PMID- 12624316 TI - Early major ischemic changes on computed tomography should preclude use of tissue plasminogen activator. PMID- 12624317 TI - Early major ischemic changes on computed tomography should not preclude use of tissue plasminogen activator. PMID- 12624318 TI - CT screening for thrombolysis: uncertainties remain. PMID- 12624319 TI - Routine or selective carotid artery shunting for carotid endarterectomy (and different methods of monitoring in selective shunting). PMID- 12624421 TI - Cre/lox: one more step in the taming of the genome. AB - Cre recombinase has become an important tool in the precise manipulation of the genome, and its adoption has led to the development of increasingly accurate mouse models for the understanding of gene function. Although much of current work exploits the alacrity and precision with which Cre catalyzes excisive DNA recombination, Cre also is adept at the insertion of heterologous DNA into the genome. The precision and efficiency with which Cre can target DNA to a predesignated locus in the genome promises to facilitate understanding of mutant genes and allelic variants in their natural chromosomal context. PMID- 12624422 TI - Gene targeting in the mouse: advances in introduction of transgenes into the genome by homologous recombination. AB - The ability to stably introduce genes into the germline of animals provides a powerful means to address the genetic basis of physiology. Introduction of genes to generate transgenic animals has facilitated the development of complex genetic models of disease, as well as the in vivo study of gene function. However, one drawback of traditional transgenic technologies in which genes are microinjected into early-stage embryos is that there is little control over where and in how many copies genes are introduced into the genome. The development of animal transgenic technologies, which take advantage of homologous recombination mechanisms and the manipulation of embryonic stem (ES) cells, allows investigators to target and alter specific loci. In mouse transgenic systems, a plethora of sophisticated gene-targeting strategies now permit investigators to manipulate the genome in ways that essentially allow one to introduce virtually any desired change into the genome. Furthermore, when coupled with systems that allow for conditional gene expression, these gene-targeting strategies allow both temporal and tissue specific control of alterations to the genome. In the present review we briefly discuss some of the more recent gene-targeting strategies that have been developed to address the limitations of traditional animal transgenesis. PMID- 12624424 TI - A model for tissue-specific inducible insulin-like growth factor-I (IGF-I) inactivation to determine the physiological role of liver-derived IGF-I. AB - Insulin-like growth factor-I (IGF-I) has important growthpromoting and metabolic effects and is expressed in virtually every tissue of the body. The highest expression is found in the liver, but the physiological role of liver-derived IGF I is unknown. It has been difficult to separate the endocrine effects of liver derived IGF-I from the autocrine/paracrine effects of locally produced IGF-I in peripheral tissues. Therefore, we have developed a mouse model with a liver specific inducible deletion of the IGF-I gene (LI-IGF-I-/- mouse). The LI-IGF-I-/ mouse has dramatically reduced (>80%) serum IGF-I levels, demonstrating that the major part of serum IGF-I is liver-derived. Surprisingly, LI-IGFI -/- mice demonstrate a normal appendicular skeletal growth up to at least 12 mo of age despite the dramatic decrease in circulating IGF-I levels, indicating that liver derived IGF-I is not required for appendicular skeletal growth. However, the adult axial skeletal growth is reduced in the LI-IGF-I-/- mice. Furthermore, the amount of cortical bone is reduced due to decreased radial growth of the cortical bone, while the trabecular bone mineral density is unchanged in the LI-IGFI -/- mice. The decreased levels of circulating IGF-I are associated with increased serum levels of growth hormone (GH), indicating a role for liver-derived IGFI in the negative-feedback regulation of GH secretion. Measurements of factors regulating GH secretion in the pituitary and in the hypothalamus revealed an increased expression of GH-releasing-hormone (GHRH) and GHsecretagogue (GHS) receptors in the pituitary of LI-IGFI -/- mice. This in turn results in an increased sensitivity to systemically administered GHRH and GHS, demonstrating that the regulatory action of liver-derived IGF-I on GH secretion is at the pituitary rather than at the hypothalamic level. The liver is an important metabolic organ and LI-IGF-I-/- mice are markedly hyperinsulinemic and yet normoglycemic, consistent with an adequately compensated insulin resistance. Interestingly, LI-IGF-I-/- mice display a reduced age-dependent fat mass accumulation compared with control mice. Furthermore, LI-IGF-I-/- mice have increased blood pressure attributable to increased peripheral resistance indicating a role for liver-derived IGF-I in the regulation of blood pressure. In conclusion, liver-derived IGF-I is important for carbohydrate and lipid metabolism and for the regulation of GH secretion at the pituitary level. Furthermore, it regulates adult axial skeletal growth and cortical radial growth while it is not required for appendicular skeletal growth. PMID- 12624423 TI - The role of circulating IGF-I: lessons from human and animal models. AB - Insulin-like growth factors (IGF-I and IGF-II) play a crucial role in regulating cell proliferation and differentiation. The IGFs have mitogenic and antiapoptotic effects on normal and transformed cells. These peptide growth factors are produced by virtually all tissues and act in an endocrine, autocrine, and paracrine fashion. The endocrine form of IGF-I originates mostly (75%) from the liver and IGF-binding proteins regulate its bioactivity. Compared to other peptide growth factors, the IGFs are in abundant supply in circulation. The role of this large reservoir of IGFs has been debated for many years. In the last few years substantial progress has been made in understanding the function of the endocrine IGF-I using new animal models. This review will revisit the IGF system with particular attention to the role of circulating IGF-I in growth regulation, metabolism, and cancer. PMID- 12624425 TI - Tissue-specific targeting of the insulin receptor gene. AB - The techniques to study the mechanisms that underlie the pathogenesis of disease processes have been revolutionized by the development of methods that allow spatiotemporal control of gene deletion or gene expression in transgenic and knockout animals. The ability to interfere with the function of a single protein in a specific tissue allows unprecedented flexibility for exploring gene function in both health and disease. The present review will summarize some of the different knockouts and transgenics generated recently to study type 2 diabetes and critically evaluate the techniques used to examine the function of the insulin receptor in two nonclassical insulin target tissues--the pancreatic islet and the central nervous system. PMID- 12624427 TI - Glucagon-like peptide 1 content of intestinal tract in adult rats injected with streptozotocin either during neonatal period or 7 d before sacrifice. AB - Glucagon-like peptide 1 (GLP-1) content of the intestinal tract was recently found to be lower in diabetesprone BioBreeding (BBdp) rats than in the corresponding control animals (BBc rats), a finding compatible with the idea that an inflammatory intestinal state precedes insulitis in these diabetes-prone animals. This study aimed at measuring GLP-1 content of the intestinal tract both in another animal model of type 1 diabetes and in an animal model of type 2 diabetes. GLP-1 content of the jejunum, ileum, colon, and cecum was measured in male and female adult control rats and animals injected with streptozotocin (STZ) either during the neonatal period or 7 d before sacrifice. GLP-1 content of the intestinal tract was higher in type 1 diabetic rats than in control animals. Such was not the case in the type 2 diabetic rats. The findings recorded in the rats injected with STZ either during the neonatal period or later in life indicate that hyperglycemia and/or insulin deficiency do not cause a decrease in GLP-1 content of the intestinal tract. On the contrary, such a content may increase when the glucose intolerance and hypoinsulinemia are sufficiently pronounced, as was the case in the type 1 diabetic rats. These findings are thus compatible with the view that the decreased GLP-1 content of the intestinal tract in BBdp rats may result from intestinal inflammation. PMID- 12624426 TI - Pancreatic cell lineage analyses in mice. AB - Considerable knowledge of the ontogeny of the endocrine pancreas has been gained in recent years, mainly through the use of two complementary genetic approaches in transgenic mice: gene inactivation or overexpression (to assess gene function) and genetic labeling of precursor cells (to determine cell lineages). In recent years, in vivo Cre/loxP-based direct cell tracing experiments show that (i) all pancreatic cells differentiate from pdx1-expressing precursors, (ii) p48 is involved in the exocrine and endocrine pancreatic lineages, (iii) islet endocrine cells derive from ngn3-expressing progenitor cells, and (iv) insulin cells do not derive from glucagon- expressing progenitors. Lineage analyses allow the identification of progenitor cells from which mature cell types differentiate. Once identified, such progenitors can be labeled and isolated, and their differentiation and gene expression profiles studied in vitro. Understanding pancreatic cell lineages is highly relevant for future cell replacement therapies in diabetic patients, helping to define the identity of putative (endodermal) pancreatic stem cells. PMID- 12624428 TI - Role of phosphatidylinositol 3-kinase/Akt pathway in angiotensin II and insulin like growth factor-1 modulation of nitric oxide synthase in vascular smooth muscle cells. AB - The aim of this study was to examine the role of the phosphatidylinositol 3 kinase (PI3K)/serine/threonine kinase Akt signaling pathway in mediating interactions between angiotensin II (Ang II) and insulin-like growth factor-1 (IGF-1) in regulation of inducible nitric oxide synthase (iNOS) in vascular smooth muscle cells (VSMCs). Exposure to 100 nM IGF-1 for 10 min resulted in increased insulin-receptor substrate-1 associated PI3K activity and Akt kinase activity, whereas 100 nM Ang II pretreatment for 5 min strikingly decreased these IGF-1 effects. NOS activity was also increased in VSMCs following exposure to IGF 1 (10 min up to 24 h). Pretreatment with Ang II for 5 min reduced IGF-1-induced NOS activity. IGF-1 treatment for 24 hr increased iNOS gene transcription, and Ang II pretreatment reduced this stimulation of iNOS gene expression by attenuating PI3K/Akt signaling. These results implicate PI3K/ Akt pathways in Ang II/IGF-1 regulation of iNOS in VSMCs. PMID- 12624429 TI - Invertase, maltase, lactase, and peroxidase activities in duodenum of BB rats. AB - The development of immune-mediated diabetes in BB rats may involve a defect of the gastrointestinal tract (GI), as suggested by increased gut permeability. This study aimed at measuring invertase, maltase, lactase, and peroxidase activities in the duodenum of diabetesprone BioBreeding (BBdp) rats and control BioBreeding rats (BBc) given free access to NIH-07 diet up to the time of killing at 60 66 d of age. After washing the entire small intestine, the duodenal mucosa was scraped off in the first 5-cm segment from the pylorus and frozen in distilled water. Invertase, maltase, and lactase activities were measured by monitoring the conversion of [U-(14)C]sucrose, [U-(14)C]maltose, and [D-[1-(14)C]glucose] lactose to radioactive hexoses, which were phosphorylated in the presence of adenosine triphosphatase and yeast hexokinase and then separated from their precursor by ion-exchange chromatography. Peroxidase activity was measured by a spectrophotometric procedure. In the BBdp rats, the activity of invertase, maltase, and lactase averaged, respectively, 70.2 +/- 4.4, 81.2 +/- 4.3, and 75.7 +/- 4.1% (n = 16 and p < 0.001 in all cases) of the control values found in BBc rats of the same sex. Inversely, after exclusion of two female BBc rats with abnormally high plasma D-glucose concentration, the activity of peroxidase in the BBdp rats averaged 157.4 +/- 20.0% (n = 16; p < 0.02) of the mean control value recorded in BBc rats of the same sex (100.0 +/- 9.3%; n = 14). These findings are compatible with the view that a proinflammatory state of the GI associated with compromise function may precede the occurrence of pancreatic insulitis in BBdp rats and, possibly, human subjects with type 1 diabetes. PMID- 12624430 TI - Effect of ethanol on regulation of (Na + K)-adenosine triphosphatase by aldosterone and dexamethasone in cultured renal papillary collecting duct cells. AB - Chronic ethanol exposure causes alterations in biologic membranes of different cell types. (Na + K) adenosine triphosphatase (ATPase), a membrane-bound enzyme inhibited by the acute presence of ethanol, increases its activity in rat kidney after chronic ethanol consumption. The aim of this investigation was to evaluate the effect of ethanol on the modulation of (Na + K)-ATPase by glucocorticoids and mineralocorticoids in renal papillary collecting duct cells. Cultured renal papillary collecting duct cells were exposed to a medium containing 150 mM ethanol plus either 100 nM aldosterone or 10 nM dexamethasone. Control groups were cultured in the absence of ethanol and/or the hormones. Mg(2+)-ATPase was used as control enzyme. The activity of ATPases was measured by ATP hydrolysis. Ethanol increased the activities of (Na + K)-ATPase and Mg(2+)- ATPase in 29 and 33% of controls, respectively; only (Na + K)-ATPase activity was elevated in the presence of aldosterone or dexamethasone, whereas Mg(2+)-ATPase was unaltered by these hormones. The effects of aldosterone and dexamethasone on (Na + K)-ATPase activity were augmented by ethanol in 50 and 19% of controls, respectively. These results suggest that ethanol treatment enhances the upregulation of (Na + K) ATPase activity by both aldosterone and dexamethasone, in cultured renal papillary collecting duct cells. PMID- 12624431 TI - The expression of angiotensin and endothelin system members in bovine corpus luteum during estrous cycle and pregnancy. AB - The aim of this study was to determine the changing profiles of the mRNA expression of members of angiotensin and endothelin system in bovine corpus luteum (CL) from different stages of the estrous cycle and pregnancy. Corpora lutea were accordingly assigned to the following stages; d 1 2, 3 4, 5 7, 8 12, 13 18, >18 (after regression) of estrous cycle and of early and late pregnancy (<4 and >4 mo). The block RT-PCR analysis of CL showed a significantly higher angiotensin converting enzyme (ACE) mRNA expression during mid and late luteal phases as well as after regression, but lower levels during pregnancy. Full quantitative real-time RT-PCR (LightCycler) confirmed this pattern of ACE mRNA expression. The angiotensin receptor type 1 (AT1R) mRNA expression was relatively stable throughout the periods examined. In contrast, AT2R mRNA temporarily decreased on d 8 12, followed by an increase to the highest levels during late luteal phase, and it remained at high levels during regression and pregnancy. Concentration of angiotensin II (Ang II) peptide in luteal tissue was highest after ovulation (d 1 2), decreased afterward, increased again during late luteal phase, and decreased to lower levels during regression and pregnancy. The mRNA expression and peptide concentration of endothelin 1 (ET-1) was high after ovulation followed by a decrease during mid and late luteal phases and increased again to the highest level after regression. The endothelin receptor type B (ETR B) mRNA expression increased during late luteal phase and further after regression. In contrast, ETR-A and endothelin converting enzyme 1 (ECE-1) mRNA expression were relatively constant during all stages examined. In conclusion, the regulatory changes of both angiotensin and endothelin family members during early luteal phase and again during late luteal phase suggest a possible modulatory role of these vasoactive peptide families for bovine CL formation and regression. PMID- 12624432 TI - Inhibition of prolactin secretion from the male rat anterior pituitary by cryptic sequences of prothyrotropin releasing hormone, ProTRH178-199 and ProTRH186-199. AB - Previous studies have shown that intronic peptide sequences in the prohormone for thyrotropin-releasing hormone (TRH) have physiological actions on pituitary hormone secretion. The aim of this investigation was to examine the effect of the cryptic peptides, prothyrotropin- releasing hormone(178-199) (ProTRH(178-199)) and ProTRH(186-199), on prolactin (PRL) release from the anterior pituitary. Perifusion studies were performed with anterior pituitaries obtained from individual adult male Sprague Dawley rats at 70 90 d of age. Perifusate was collected in 5-min fractions for 25 min prior to peptide administration and for 60 min afterward. Pituitaries were perifused with a single 5 min pulse of either 2, 10, or 40 nM concentrations (peak pulse) of each peptide or the vehicle. Sixty minutes after peptide administration, a 200 mM pulse of potassium chloride was delivered to check tissue viability. Prolactin was measured in the perifusate by radioimmunoassay. Results showed that both peptides induced a significant long term suppression of prolactin secretion that was still evident at 60 min after peptide exposure. ProTRH(186-199) was similar to ProTRH(178-199) in suppressing prolactin release at the 2 and 40 nM dose, suggesting that the amino acid sequence necessary for prolactin inhibition is contained within the smaller peptide fragment. These data indicate that a cryptic sequence within the proTRH peptide can have biological activity at the level of the anterior pituitary gland in regulating prolactin secretion. PMID- 12624433 TI - Inherited adrenocorticotropin-independent macronodular adrenal hyperplasia with abnormal cortisol secretion by vasopressin and catecholamines: detection of the aberrant hormone receptors on adrenal gland. AB - ACTH-independent macronodular adrenal hyperplasia (AIMAH) is a rare disorder and an unusual cause of Cushing s syndrome, of which familial transmission has rarely been reported. In this study, a mother and her son, the former affected with definite AIMAH and the latter with possible AIMAH, are described. Although the mother manifested overt Cushing s syndrome, her son remained with no stigmata of Cushing s syndrome except for bilateral adrenal tumor and mild hypertension, and a full suppression of plasma cortisol by lowdose dexamethasone was observed in him. Recently, aberrant expression of adrenal receptors for various ligands has been noted in AIMAH patients. In our cases, provocation tests in vivo suggested that AVP and catecholamines promoted cortisol production through V1a and/or V1b receptors and via beta-adrenergic receptor, respectively. Reverse transcriptional PCR analysis of the operated adrenal tissues of mother revealed the abnormal expression of mRNA of receptors for V1b, V2, and LH/hCG, none of which was observed in a normal control. Inherited AIMAH is very rare, and the son might be at the earliest developmental stage of AIMAH among the cases reported so far. An intervention could be tried to prevent the development of overt Cushing s syndrome by suppression of the possible endogenous ligands or by blockade of the receptors that may be aberrantly expressed in his adrenal glands. PMID- 12624434 TI - Effect of high-fat diet on body composition and hormone responses to glucose tolerance tests. AB - To determine potential hormonal mediators of the effect of high-fat diets on the development of insulin resistance, blood leptin, growth hormone (GH), glucose, and insulin responses to a 2 g/kg BW oral glucose challenge were evaluated in weanling female Sprague Dawley rats that were randomly assigned to a high-fat (HF, 39% of calories, 20% fat by weight; n = 10) and moderate-fat (MF, 22% of calories, 10% fat by weight; n = 10) diets. Oral glucose challenges were administered following 5, 7, and 9 wk on the feeding trial. Animals were provided diet in excess of their requirements for growth. Body mass analysis was conducted by dual X-ray absorptiometry (DXA) on the 6th, 8th, and 10th weeks of the trial. HF animals gained more weight after 7 wk, had greater body fat than the MF animals, and similar glucose responses to the oral glucose challenges. HF rats secreted more insulin and leptin compared to MF animals. Lean body mass and serum GH and IGF-I concentrations were not different between the groups. Results of this study demonstrate that leptin but not GH or IGF-I is involved in the development of insulin resistance in growing rats as a result of excess energy intake in the form of dietary fat. PMID- 12624435 TI - Prediction of tyrosine sulfation in seven-transmembrane peptide receptors. AB - Posttranslational modification by tyrosine sulfation regulates many important protein protein interactions and modulates the binding affinity and specificity of seventransmembrane peptide receptors. We developed a log-odds position specific-scoring-matrix (PSSM) to accurately predict tyrosine sulfation using 62 tyrosine sites known to be sulfated and 421 tyrosine sites known not to be sulfated. We predict that 49 tyrosines of 32 seven-transmembrane peptide receptors are sulfated. Although we did not incorporate characteristics of confirmed sulfation sites such as clustering and conservation across species into our PSSM, our predicted sites nevertheless exhibited these characteristics. The observed conservation suggests that there are strong evolutionary pressures to preserve selected biological activity of seven-transmembrane receptors. The predicted tyrosine sulfation sites predominantly occur in the extracellular tail and extracellular loop 2, regions consistent with their association with binding pockets of the receptor. PMID- 12624473 TI - [On the path to complete health]. PMID- 12624436 TI - Castration reduces mRNA levels for calcium regulatory proteins in rat heart. AB - Sex-related differences in the cardiac phenotype have been well established. This study was designed to determine whether androgens regulate myocardial gene expression and play a role in the sex-related differences in the myocardial phenotype. Gonadectomized male rats were treated with testosterone, and myocardial gene expression was examined in whole heart using quantitative real time PCR. Gonadectomy produced a substantial decrease in mRNA levels for the androgen receptor, Na(+)/Ca(2+) exchanger, L- type calcium channel, and beta(1) adrenergic receptor (beta(1)AR). Supplementation of testosterone in castrates produced a fivefold increase in androgen receptor mRNA levels. Testosterone treatment of castrates produced almost a sixfold increase in Na(+)/Ca(2+) exchanger mRNA, a tenfold increase in Ltype calcium channel mRNA accumulation, and a fourfold increase in beta(1)AR mRNA levels. Increased calcium channel expression, beta(1)AR expression, and Na(+)/Ca(2+) exchanger expression together may alter cytosolic calcium. These results provide the first evidence that testosterone regulates expression of myocardial calcium regulating genes and thus may play a role in modulating the cardiac phenotype in males. PMID- 12624474 TI - Effects of one week juice fasting on lipid metabolism: a cohort study in healthy subjects. AB - OBJECTIVE: We investigated the effects of a popular modified juice fasting program on lipid metabolism. VOLUNTEERS AND METHODS: 5 healthy, nonobese, male volunteers fasted for 8 days. Daily energy intake was limited to 150-300 kcal/d solely as carbohydrates (vegetable and fruit drinks). Physical activity was maintained as before. At baseline, on days 2, 3, and 8 during fasting, and on days 2 and 8 after fasting, serum lipids, lipoproteins, and insulin were investigated. RESULTS: Juice fasting resulted in bi-phasic changes: Until day 2 and 3 triacylglycerols (TG), very low-density lipoprotein apolipoprotein B (VLDL apo B), and insulin decreased by 52, 51, and 65% respectively, while nonesterified fatty acids (NEFA), low-density lipoprotein (LDL) apo B, and LDL cholesterol increased by 363, 38, and 35%. Between day 3 and 8 NEFA increased; TG and insulin increased as well, but remained below baseline values, and LDL cholesterol normalized. After 8 days juice fasting significant changes (p < 0.05) compared to the baseline were found only for free cholesterol (-10%), phospholipids (-14%), apo AI (-9%), apo AII (-11%), insulin (-42%), C-peptide ( 57%), and NEFA (+535%, p = 0.0001). Total cholesterol decreased by 9% (n.s.) after 8 days. One week after the ending of fasting all parameters returned to normal. CONCLUSION: Contrary to total fasting and fasting with limited physical activity, 8 days juice fasting without limitation of physical activity results in a decrease of free cholesterol and an only initial increase of LDL cholesterol. After 8 days insulin, TG, and VLDL are still lower than at baseline, however, they have increased compared to the initial phase, probably counterregulatory to a further increase of NEFA. PMID- 12624475 TI - [The vitality of obese patients after a weight reduction by fasting]. AB - INTRODUCTION: The objective documentation of the positive results of natural medicine or physiotherapy gains raising importance in medicine. OBJECTIVE: The aim of the study was to investigate the application of the vitality test according to Pothig in the objective holistic evaluation of treatment by natural and physical medicine. SUBJECTS AND METHODS: In 48 obese patients (28 males and 20 females) with a body mass index of 32.1 to 56.5 kg/m(2) a weight reduction therapy was carried out for 21 days (16-25 days) by means of fasting according to Buchinger, accompanied by physiotherapeutic treatments. At the beginning and at the end all patients underwent a multidisciplinary functional diagnosis by means of the vitality test according to Pothig and a bioelectrical impedance analysis. On the basis of the 47 measured parameters of the entire body function (physical, mental and emotional/social) the Vitality Index and the Functional Age Index (FAI) were calculated. RESULTS: The reduction of body weight was 9.1 +/- 2.4 (5.1 15.0) kg, the reduction of body fat 5.5 +/- 1.3 (3.3-8.5) kg. At the beginning 31% of the patients showed a reduced vitality, i.e. the FAI was at least 5 years (maximum 16 years) higher then the real age. 56% of the patients matched the real age and in 13% of the patients the FAI was better with respect to the corresponding age group. During the treatment 94% of the patients showed an improvement of their FAI by 5 +/- 3 years on average (maximum 12 years). CONCLUSIONS: With the help of the presented multidisciplinary vitality test it is possible to objectively document the positive results of natural medicine and physical medicine on the whole human organism. PMID- 12624476 TI - Effects of zinc histidine and zinc sulfate on natural human keratinocytes. AB - BACKGROUND: Zinc salts are widely used as food supplements and medicinal mineral supplementation. Zinc deficiency is associated with impaired skin conditions. The influence of zinc on skin functionality has been proven in clinical investigations. OBJECTIVE: Within the following study comparative in vitro experiments were performed to study the influence of zinc sulfate (ZnSO(4)) and zinc histidine (Zn(His)(2)) on the physiology of cultured natural human keratinocytes. METHOD: Proliferation of natural human keratinocytes was quantitatively assessed by measurement of 5-bromo-2-deoxyuridine (BrdU) incorporation against an oligomeric procyanidin as positive control. Differentiation was determined by monitoring involucrin formation. Cell viability and cytotoxicity were assessed by dimethylthiazolyldiphenyltetrazolium bromide (MTT) testing and quantification of lactate dehydrogenase. RESULTS: Neither keratin synthesis as a late marker of cell differentiation nor mitochondrial cell activity were influenced by either zinc compound. The synthesis of involucrin, an early marker of differentiation, was significantly increased by both zinc salts, ZnSO(4) being the more potent stimulator. Both zinc salts significantly increased cell proliferation, with the histidine complex being more potent. Effects were in the range of the positive control. Necrotic cell toxicity decreased significantly when Zn(His)(2) was added to the cells. CONCLUSION: Under in vitro conditions Zn(His)(2) is a strong proliferation inductor of keratinocytes with a better tolerability and a lower induction of differentiation behavior than ZnSO(4). PMID- 12624477 TI - [Literature review of the latest research results concerning the positive effect of exercise therapy in chronic heart insufficiency]. AB - Scientific investigations indicate similarities in the pathophysiology of heart insufficiency and that of physical inactivity: similar changes in peripheral hemodynamics (increased peripheral vascular resistance, worsening of oxygen utilization during exercise), in autonomic control (activation of neurohumoral compensatory mechanisms, e.g. the renin-angiotensin system, overactivation of the sympathicus, reduction of vagal tonus, reduced pressosensitivity), in functional activity (reduced exercise tolerance and reduced maximum oxygen uptake), in skeletal muscle (decrease in mass, changes in structure), and in the psychological state (reduction in activity and feeling of well-being). In several, although small-scale studies it could be shown that patients with advanced left ventricular failure were able to take part in training programs without experiencing any ill effects, and that there was a positive shift in the usual typical effects of physical training, such as increase of heart rate, change in respiratory frequency, and maximum oxygen uptake. It could be shown that exercise therapy can result in a shift in the balance between the sympathetic and the parasympathetic tonus in the low- and high-frequency maxima of the R-R interval variability. The pre-training general predominance of the sympathetic tonus over the vagal tonus was changed dramatically by the training, leading to a predominance of the vagal tonus. Recent controlled studies with a randomized and controlled cross-over design and the application of a training program which was carried out regularly and independently have confirmed the positive effect of aerobic fitness training in cases of heart disease. At the end of the exercise phase, the patients experienced a significant improvement of the symptoms of left ventricular failure and of their capacity for exercise; furthermore, the training altered parts of the neurohumoral activation, which count as the main factors in the progression and death rate of patients with chronic cardiac disease. PMID- 12624478 TI - [How holistic is health?--from Hippocrates and Plato to a future sociopsychosomatic situational therapy]. AB - To be with others is helpful for the recovery from illness. It has been shown that this is true not only in human relations but also, for instance, in relating oneself to trees as well. This implies that health is not an individual but a social property. Like in Hippocratic and Platonic medicine illness means to have got out of oneself or out of one's nature in the widest - even cosmic - sense. The basic task of complementary and natural medicine is to prevent just this; to cure diseases can only be second-class. PMID- 12624479 TI - [Effectiveness of herbal drugs]. PMID- 12624480 TI - Dehydroascorbic acid in urine as a possible indicator of surgical stress. AB - BACKGROUND/AIM: L-Ascorbic acid (AA) is the predominant circulating form of vitamin C found in human blood. It has been hypothesized that surgical stress increases the vitamin C metabolite dehydroascorbic acid (DHAA). Vitamin C is mainly excreted through the kidneys. In this study, the ratio of AA to DHAA excreted in urine was determined in patients who had undergone total hip joint endoprosthesis surgery (n = 12), and the results were compared with data obtained from healthy controls (n = 12). METHODS: All subjects received 1,000 mg sodium ascorbate intravenously three times a day (every 8 h) for 8 days, starting 2 days prior to surgery. Total urine was collected daily while subsequent determinations of AA and DHAA were performed photometrically. RESULTS: Administration of vitamin C led to average daily excretions of the combined products AA + DHAA of 2,343 +/- 438 mg/day (mean value +/- confidence intervals). The initial average ratio DHAA/AA of all 24 probands was 0.064 (6% DHAA; 153 +/- 76 mg/day). One day after surgery, an increase in the DHAA/AA ratio to 0.165 (15% DHAA; 332 +/- 107 mg/day) was measured in the patients. The ratio decreased 2 days after surgery and returned to normal within 5 days. CONCLUSION: Our data indicate that surgery increases the oxidation of AA and urinary excretion of DHAA, as a result of the enhanced formation of free radicals. PMID- 12624481 TI - Effect of different doses of vitamin E on the incidence of malformations in pregnant diabetic rats. AB - BACKGROUND/AIMS: Previously we have shown that administration of 150 mg of vitamin E (alpha-tocopherol) per day to rats having diabetes decreases the rate of embryo malformations and increases their maturation and size. The present study was addressed to determine the effects of different doses of vitamin E upon these parameters. METHODS: Female rats were made diabetic (D) with streptozotocin, and from day 0 of gestation they were treated daily with 25 (D+25), 50 (D+50), 100 (D+100), 150 (D+150), and 500 (D+500) mg of vitamin E administered orally and were compared with control (C) animals. RESULTS: On day 11.5 of gestation, crown-rump length, somite number, and protein and DNA levels were lower in D than in C embryos. Crown-rump length and somite number increased with 100 mg or higher doses of vitamin E, although the values observed in C embryos were not reached. The proportions of reabsorption and malformations were 24.7 and 50%, respectively, in D rats, and in the rats supplemented with vitamin E they decreased to 22.7 and 19% in D+25, 16.4 and 21.3% in D+50, 16.2 and 12% in D+100, 12.9 and 13.9% in D+150, and to 43.9 and 10.8% in D+500 rats, whereas the values were 6.8 and 4.9% in C animals. CONCLUSIONS: Administration of vitamin E to D rats decreases the rate of embryo malformations, dependent on the dose administered. However, high doses have a negative effect in the conceptus, as shown by the increased rate of reabsorptions in the D+500 group. PMID- 12624482 TI - Creatine supplementation affects glucose homeostasis but not insulin secretion in humans. AB - AIMS: In this study, it was investigated whether the glucose homeostasis is affected by dietary creatine supplementation. For this purpose, the plasma glucose concentration and the plasma insulin response to an oral glucose load were measured in creatine-supplemented vegetarians. METHODS: The subjects were supplemented with either 5 g of creatine monohydrate (creatine-treated group, CREAT) or 5 g of maltodextrin (control group, CON) per day for 42 days. On days 0 and 43, blood samples were collected before as well as 10, 20, and 30 min following an oral glucose load and analyzed for plasma creatine, insulin, and glucose levels. RESULTS: Creatine supplementation resulted in an increase in plasma creatine (CREAT 92.7 +/- 14.6 micro M vs. CON 31.2 +/- 3.2 micro M; p = 0.001). There was a trend (p = 0.07) towards elevated fasting plasma glucose levels following creatine supplementation, while the plasma glucose response to the glucose load was enhanced (CREAT 168.2 +/- 5.3 mM. min vs. CON 129.6 +/- 14.7 mM.min; p = 0.05). There was no difference observed in the plasma insulin response to the glucose load between the groups. CONCLUSION: This study shows that creatine supplementation may result in abnormalities in glucose homeostasis in the absence of changes in insulin secretion. PMID- 12624484 TI - Metabolism of long-chain polyunsaturated fatty acids in rat kidney cells. AB - The metabolism of long-chain polyunsaturated fatty acids is characterized in tissues, such as liver and heart, especially from studies based on isolated cells incubated with radiolabelled fatty acid substrates. Differently, only little is known about the metabolism of fatty acids in the kidney. It is controversial whether the kidney possesses the ability to desaturate long-chain fatty acids or whether kidney cells are dependent on performed polyunsaturated fatty acids transported from the liver. In this study we used isolated rat kidney cells obtained by a perfusion technique. The cells were incubated with [1-(14)C] labelled 18:3(n-3) or 20:3(n-6) fatty acids which were incorporated into complex lipids or desaturated/elongated. The lipids were separated by thin-layer chromatography and high-performance liquid chromatography. The present study shows that isolated kidney cells take up and esterify labelled long-chain polyunsaturated fatty acids in a time- and concentration-dependent manner. We have also demonstrated that isolated rat kidney cells only to a minor extent Delta6-desaturate labelled 18:3(n-3) to 18:4 (n-3). Conversely, the Delta 5 desaturation of 20:3(n-6) to 20:4(n-6) is far more active. It may thus be concluded that the kidney, at least in part, must obtain its C(20) and C(22) fatty acids from the circulation, while the active Delta5-desaturase suggests that preformed C(20) fatty acids can be converted to more unsaturated homologues in the kidney. PMID- 12624483 TI - Relative bioavailability and antioxidant potential of two coenzyme q10 preparations. AB - Coenzyme Q10 (CoQ10) is synthesized by the human body and found in certain foods. Daily supplementation of CoQ10 could protect against heart disease but the bioavailability of CoQ10 supplements depends on the formulation taken. We compared the bioavailability and antioxidant properties of two commercial CoQ10 formulations, a commercial grade CoQ10 powder (commercial grade CoQ) and a new BT CoQ10 BIO-TRANSFORMED (BT-CoQ10) obtained by fermentation of a soy-based, CoQ10 rich media with baker's yeast. Eleven healthy individuals participated in a randomized two-way crossover trial, with a 3-week washout period. Capsules containing 300 mg of either BT-CoQ10 or commercial grade CoQ10 were given daily for 1 week and multiple blood samples were taken for CoQ10, glutathione and glutathione peroxidase (GPx) determination. In 3 subjects, baseline plasma CoQ10 levels were lower prior to BT than prior to commercial grade CoQ treatment. In the remaining participants, ingestion of BT vs. commercial grade CoQ significantly increased maximum plasma CoQ10 concentration (+126%, p = 0.04) and tended to increase CoQ10 area under the curve from 0 to 24 h (+160%, p = 0.07). One week of treatment with each formulation increased plasma CoQ10 but did not alter plasma glutathione or GPx activity. The enhanced bioavailability of the BT product might be due to its predominantly reduced, hydrophilic membrane-complex form. PMID- 12624485 TI - Influence of dietary linoleic acid intake with different fat intakes on arachidonic acid concentrations in plasma and platelet lipids and eicosanoid biosynthesis in female volunteers. AB - BACKGROUND/AIM: N-6 fatty acids are considered to promote diseases prevalent in industrialized countries and characterized by an increased eicosanoid biosynthesis from arachidonic acid (AA). We investigated the impact of the linoleic acid (LA) intake on AA levels in humans. METHODS: Six healthy female volunteers (age range 23-34 years) were given liquid formula diets (LFD) devoid of AA for 6 weeks, providing a constant intake of zero energy% (LFD 0: protein 15%, carbohydrates 85%) or 20 energy% (LFD 20: protein 15%, carbohydrates 55%, fat 30%) LA, for 3 weeks each. Fatty acids of plasma cholesteryl esters and platelet lipids were determined each week, and the prostaglandin biosynthesis was measured in 24-hour urine samples. RESULTS: LFD 0 increased (+31% of initial value) and LFD 20 lowered (-30% of initial value) the percentage of AA in plasma cholesteryl esters and platelet lipids. Moreover, absence of dietary AA lowered the percentages of AA in plasma (-31% week 0 vs. week 6) and platelet (-11%) lipids, indicating a low transformation of LA. LFD 0 reduced urinary metabolite levels of prostaglandins D, E, and F in 24-hour urine samples (-48%, p < 0.001) within 24 h, but did not significantly affect platelet aggregation (-10%) and thromboxane formation (-25%). LFD 20 significantly lowered platelet aggregation ( 25%) and thromboxane formation (-43%). The prostaglandin metabolite levels increased during the first 10 days, declined thereafter, and were lower than the preexperimental values at the end of the 3-week period. CONCLUSIONS: The results show that dietary LA does not increase the AA levels in plasma or platelet lipids and does not persistently contribute to prostaglandin biosynthesis which is increased by AA intake with Western diets. PMID- 12624486 TI - Quantity and quality of dietary fat, carbohydrate, and fiber intake in the German EPIC cohorts. AB - AIM: This evaluation aims to describe the quantity and quality of dietary fat, carbohydrate and fiber intake in both German cohorts participating in the European Prospective Investigation into Cancer and Nutrition (EPIC). METHODS: Estimates are based on standardized computer-guided 24-hour dietary recalls from 1,078 women and 1,013 men in Heidelberg and 898 women and 1,032 men in Potsdam. In a subsample, plasma phospholipid (PL) fatty acids were analyzed as well. RESULTS: Adjusted mean dietary intake estimates demonstrated that the contribution of fat as well as n-6 and n-3 polyunsaturated fatty acids (PUFA) to the total daily energy intake was higher in both women and men of EPIC-Potsdam compared to EPIC-Heidelberg. Surprisingly, the dietary n-6/n-3 PUFA ratio was lower in the Potsdam cohort. These results were confirmed by means of the PL fatty acid pattern. Besides the higher contribution of polysaccharides to total energy intake in EPIC-Heidelberg, women of the Heidelberg cohort revealed a significantly lower contribution of mono- and disaccharides (sucrose) to total energy intake. Although total fiber intake data were similar in both cohorts, analysis by food groups showed differences in dietary fiber intake originating from the food groups cereals, fruits and potatoes. CONCLUSION: The results demonstrate distinct differences in the dietary fat, carbohydrate and fiber intake between both German EPIC cohorts, which contribute to the exposure variation in the whole of EPIC. PMID- 12624487 TI - Uropathogenic Escherichia coli toxins induce caspase-independent apoptosis in renal proximal tubular cells via ERK signaling. AB - BACKGROUND: Pyelonephritis is a risk factor for renal tubular epithelial cell damage. Recent studies have shown that Escherichia coli and/or its toxins may stimulate apoptotic cell death in renal tubular cells, but the underlying molecular mechanisms remain to be elucidated. METHODS: Confluent LLC-PK(1) cells were exposed to E. coli toxins from overnight cultures of the uropathogenic O6K13H1 (O6) and the nonpathogenic W3110. The cell death was studied with morphological and biological assay. RESULTS: E. coli soluble toxins from uropathogenic O6:K13:H1(O6) strain were found to induce apoptosis in a dose- and time-dependent manner in LLC-PK1 cells. The expression of FasR and the phosphorylation of ERK1/2 were significantly upregulated by O6 soluble toxins in a time-dependent manner. Cell death was completely inhibited by two specific ERK1/2 inhibitors, but not by a broad caspase inhibitor, zVAD-fmk, implicating a caspase-independent pathway via ERK. Moreover, we found that lysophosphatidic acid could trigger a survival signal through G-proteins and PI3K. CONCLUSION: We demonstrate that apoptosis induced by uropathogenic E. coli toxins is dependent on ERK1/2. Caspases, although being activated, are not necessary for cell death, and they act after the ERK signaling at which point cells become committed to cell death or can be rescued. PMID- 12624488 TI - Possibility of progenitor cell mobilization during the hematological recovery following peripheral blood stem cell autograft. AB - Twenty-four patients with hematological malignancies were studied during recovery following autografting in order to establish the proportion of patients that show CD34+ cell mobilization and the kinetics of mobilized CD34-positive cells. The patients showed a peak in peripheral blood (PB) CD34+ cells after a median of 14 days (range 12-20) following reinfusion. According to the number of circulating CD34+ cells, two groups could be clearly distinguished: 17 patients (group A) with <10 PB CD34+ cells/microl (median 1.2, range 0-5) and 7 patients (group B) with >10 CD34+ cells/microl (median 51, range 13-123). Compared to group A, patients of group B showed a faster hematological reconstitution of both polymorphonuclear leukocytes >500/microl (12 vs. 15 days) and platelets >50,000/microl (12 vs. 17 days). The expression of the beta1 integrin CD49d was similar in the two groups of patients, while a lower expression of the beta2 integrin CD11a and a greater expression of the L-selectin CD62L were observed in the PB CD34+ cells of group B patients. Both in the PB and in the BM, the number of CFU-GEMM, CFU- GM, CFU-E and BFU-E of group B was significantly greater than that of group A. However, when the clonogenic potential of a single CD34+ cell was evaluated, no major differences in the number of colonies produced per CD34+ cell were found between the two groups. PMID- 12624489 TI - High incidence of hemochromatosis gene mutations in the myelodysplastic syndrome: the Budapest Study on 50 patients. AB - Genotypic testing of nonselected patients with the myelodysplastic syndrome (MDS) for the C282Y and H63D mutations of the HFE gene responsible for hereditary hemochromatosis revealed a significantly increased frequency of these mutations when compared to healthy blood donors reflecting the average population. Among the 50 patients examined [26 refractory anemia (RA), 9 refractory anemia with ring sideroblasts (RARS), 2 refractory anemia with excess of blasts (RAEB) and 13 refractory anemia with excess of blasts in transformation (RAEB-t)] there were 24 heterozygotes (20 for H63D and 4 for C282Y), 1 homozygote for H63D and 1 compound heterozygote. The difference between the HFE-positive and HFE-negative MDS patients as regards initial serum iron and transferrin saturation was not significant. Inevitably the iron overload syndrome eventually develops in MDS patients due to intrinsic characteristics of the disease as well as an escalating need for blood transfusion therapy in the course of the disease. The high incidence rate of HFE gene mutations among MDS patients may also contribute to this vicious circle. PMID- 12624490 TI - Cytokine gene expression in regenerating haematopoietic tissues of mice after cyclophosphamide treatment. AB - The goal of the study was to investigate changes in expression of selected growth factors tentatively involved in regeneration of haematopoietic tissues (bone marrow and spleen) following cyclophosphamide (CY) damage in the mouse. The bone marrow (BM) and spleen were examined separately, since the regenerating pattern for haematopoietic progenitor cells (HPC) markedly differs in these two haematopoietically active organs after CY. Cytokines assumed to have a stimulatory effect on HPC - stem cell factor (SCF), fetal liver tyrosine kinase 3 ligand (flt3-ligand), thrombopoietin (TPO), stromal cell-derived factor 1 (SDF 1), oncostatin M (OSM) -, a suppressive effect on HPC proliferation - macrophage inflammatory protein-1alpha (MIP-1alpha), transforming growth factor-beta1 (TGFbeta1), tumour necrosis factor-alpha (TNFalpha) - and to be involved in migration of HPC (SCF, flt3-ligand, MIP1alpha, SDF-1) were examined at the level of mRNA expression by means of real-time RT-PCR. The expression of a particular cytokine appears to be similar in both BM and spleen of untreated mice. CY administration changed the expression pattern of the studied genes in BM and spleen. In BM, the levels of mRNAs for SCF and SDF-1 were increased and that for TGFbeta1 decreased at time intervals at which HPC are known to proliferate intensively during BM regeneration. In contrast, stimulated proliferation of HPC in spleen was accompanied by increased expression of flt3-ligand and oncostatin M. Upon mobilization of HPC from BM into blood after CY, the expression of SCF, TPO, SDF-1 and TGFbeta1 tends to decrease in BM. Accumulation of HPC in spleen is accompanied by increased mRNA for flt3-ligand and OSM. Our findings demonstrate that different cytokines may be involved in the proliferation and mobilization/homing of HPC during recovery after CY damage in BM and spleen. PMID- 12624491 TI - Ceramide 1-phosphate formation in neutrophils. AB - In this study, we examined the metabolism of [(3)H]N-hexanoylsphingosine [C(6) ceramide (Cer)] in neutrophils, erythrocytes, platelets and mononuclear cells. [(3)H]C(6)-Cer, exogenously added and incorporated into the inside of the cell, was found to be converted to other radioactive sphingolipids in these differentiated blood cells, except erythrocytes, which were unable to metabolize C(6)-Cer. Only in neutrophils was a specific radioactive band, comigrating with a standard C(6)-Cer 1-phosphate (C(6)-Cer-1-P) on thin layer chromatography, observed in a time-dependent manner. This metabolite was confirmed to be C(6)-Cer 1-P by its sensitivity to acid treatment and resistance to mild alkaline hydrolysis. Neutrophil [(3)H]C(6)-Cer conversion into [(3)H]C(6)-Cer-1-P, reflecting Cer kinase activity, was not affected by cell stimulation. Furthermore, extracellular release of [(3)H]C(6)-Cer-1-P was not observed either. Exogenous addition of C(8)-Cer-1-P failed to induce intracellular Ca(2+) mobilization or affect the response induced by the formyl peptide. Furthermore, neutrophil morphology was not affected by C(8)-Cer-1-P. Although specific expression of Cer kinase suggests an important role for Cer-1-P in neutrophils, its functional role(s) remain to be clarified. PMID- 12624492 TI - Effect of retinoic acid treatment on cytogenetic remission of chronic myeloid leukaemia. AB - The cytogenetic responses during the first chronic phase of 11 patients with chronic myeloid leukaemia (CML) treated with all-trans retinoic acid (ATRA) + interferon (IFN) were compared with those of 9 other CML patients treated with IFN alone. Metaphase and interphase cytogenetics and a semi-quantitative polymerase chain reaction were used to evaluate the cytogenetic responses. Two of the 11 patients in the ATRA + IFN group were withdrawn, one of them because of IFN intolerance, and the other because of non-compliance. Of the 9 remaining ATRA + IFN-treated patients 6 exhibited major cytogenetic responses, 3 of which were complete. Of the 9 patients treated with IFN alone, only 2 showed major cytogenetic responses. No severe adverse effects were observed. The preliminary results suggest that the ATRA + IFN combination may be superior in achieving cytogenetic remission in the first chronic phase of CML. PMID- 12624493 TI - Severe aplastic anemia with autoimmune thyroiditis showing no hematological response to intensive immunosuppressive therapy. AB - A 39-year-old woman with severe aplastic anemia (SAA) was transferred to our institution. She also had autoimmune thyroiditis with several positive autoantibodies. Clonal or oligoclonal T-cell proliferation was demonstrated by determining the size distribution of the complementarity-determining region 3 (CDR3) of T-cell receptor beta-chain (TCR-Vbeta) subfamilies in the patient's bone marrow and peripheral blood cells. The results suggested that hematopoiesis was suppressed by immune-mediated mechanisms. Immunosuppressive therapy for SAA using cyclosporin A (CsA) alone or concurrent CsA and antithymocyte globulin (ATG) failed to induce a hematological response. The intensity of the autoantibodies, however, partially decreased during this period. In addition, the CD4/CD8 ratio was inverted after immunosuppressive therapy. These observations indicate that, in a subset of SAA, immune-mediated hematopoietic suppression cannot be successfully treated by conventional immunosuppressive therapy, even though a substantial improvement in the underlying immunological changes can be achieved. Other therapies such as hematopoietic stem cell transplantation or more intensified repeated ATG therapy may be necessary for such patients. PMID- 12624495 TI - A case of chronic myeloid leukemia complicated with minimal change nephrotic syndrome. AB - We present a 28-year-old patient with chronic myeloid leukemia (CML) in chronic phase complicated with nephrotic syndrome. The bone marrow cells revealed the presence of Philadelphia chromosome, the cytogenetic hallmark of CML, that results from a balanced, reciprocal translocation between the long arms of chromosomes 9 and 22, t(9;22)(q34;q11). This reciprocal translocation leads to the formation of the BCR/ABL fusion gene, the presence of which was confirmed using the highly sensitive fluorescence in situ hybridization technique. The renal biopsy was compatible with minimal change nephrotic syndrome. To the best of our knowledge, this is the first case of minimal change nephrotic syndrome associated with CML before the administration of any therapy. PMID- 12624494 TI - Primary NK/T cell lymphoma of the testis. A case report and review of the literature. AB - We report a case of aggressive 'nasal type' natural killer (NK)/T cell lymphoma initially presenting as a testicular tumor in a Korean man, which quickly took a fatal course by widespread dissemination. Histologically, the testicular mass showed a diffuse dense infiltrate of medium-sized and atypical large lymphoid cells with angiocentric and angiodestructive infiltration and areas of coagulative necrosis on hematoxylin-eosin stained sections. Immunophenotyping by immunohistochemistry yielded surface markers consistent with NK/T cell lymphoma. The Epstein-Barr virus genome was detected by in situ hybridization. During involved-field irradiation and chemotherapy following radical orchiectomy, the tumor disseminated shortly to the skin and soft tissue of his anterior chest wall and central nervous system (CNS). Identical lymphoid infiltrates were present in the patient's skin. CNS involvement was interpreted as having a leptomeningeal seeding. To the best of our knowledge, this is the 9th reported case of confirmed NK/T cell lymphoma arising from the testis. Relevant literature is reviewed, and the clinicopathologic features, natural history, and treatment options for primary testicular NK/T cell lymphoma are discussed. PMID- 12624496 TI - Transient elevation of platelet count in patients with chronic idiopathic thrombocytopenic purpura: association with infection. AB - Platelet count occasionally increases after infection in patients with chronic idiopathic thrombocytopenic purpura (ITP). We report 5 such patients. Three of them were males. Between November 1992 and December 2001, increased platelet counts were observed 9 times in 5 patients with chronic ITP. Corticosteroids were administered during 7 episodes; splenectomy was performed to treat 3 episodes. Antibiotics were administered in the course of 6 episodes to treat complicating infection. Platelet counts showing 1.4- to 24-fold increases have been observed. Median duration from the onset of infection until peak platelet count was 14 days. However, platelet count finally decreased to the initial level in all episodes. These findings suggest that infection is associated with the elevation of platelet count in some patients with chronic ITP, although this phenomenon is transient. PMID- 12624497 TI - Polymorphisms of glutathione S-transferase mu1 (GSTM1) and theta1 (GSTT1) genes in multiple myeloma. PMID- 12624498 TI - Treatment of angioimmunoblastic lymphadenopathy with dysproteinemia-type T-cell lymphoma with fludarabine. PMID- 12624499 TI - Classic hypoglossal-facial nerve anastomosis after acoustic neuroma resection. A review of 46 cases. AB - To clarify the factors contributing to patient satisfaction with facial movement after the classic hypoglossal-facial nerve anastomosis, we examined 46 such patients who consented to an interview and video-recording. No correlation was seen between the physician's evaluation of returned function (according to Yanagihara's 40-point scale and the grading system of House-Brackmann) and the scores (full marks: 100 points) the patients assigned to their own facial movements. Instead the scores that the patients assigned were closely related to subjective oral dysfunction, such as difficulty in masticating, articulating, and swallowing. Those scores did not correlate with eye-related functional deficits. These findings indicate that modified techniques to minimize the hypoglossal nerve deficit may enhance patient satisfaction without sacrificing recovery of facial movements. PMID- 12624500 TI - Distribution of beta-tubulin in guinea pig inner ear. AB - Our previous research had suggested that beta-tubulin might be an autoantigen for autoimmune inner ear disease. In this study, the expression of beta-tubulin in inner ears of normal and tubulin-immunized guinea pigs was examined by immunohistochemical staining. Strong immunoreactivity to beta-tubulin monoclonal antibody was found in stria vascularis, neurons of the spiral ganglion, cochlear nerve fibers and spiral ligament. Diffuse staining was found in the stria vascularis and the neurons of the spiral ganglion, while dense network staining was found in the spiral ligament, the nerve fibers and the vestibular end organs. The semicircular canals, endolymphatic duct and sac were also positively stained. In inner ears of guinea pigs challenged with beta-tubulin, staining intensity was diminished in the stria vascularis, the spiral ligament, and the neurons of the spiral ganglion. The results suggest that beta-tubulin is distributed to most structures of guinea pig inner ear. A challenge to the inner ear by tubulin could change the beta-tubulin distribution and cause degeneration in the spiral ganglion. The results support the hypothesis that beta-tubulin might be an autoantigen for autoimmune inner ear disease. PMID- 12624501 TI - TUNEL-positive labeling in mouse inner ear caused by tubulin immunization is not apoptosis. AB - Apoptosis is involved in all fundamental processes of the immune system. To study whether apoptosis plays any role in the pathogenesis of autoimmune inner ear disease, we immunized Balb/c mice with tubulin. The inner ears were examined with TUNEL (in situ terminal dUTP nick-end labeling) and immunocytochemistry of the apoptosis regulatory proteins Bcl-2 and Bax. TUNEL-positive cells are found in the tubulin-immunized inner ears but not in the control inner ears. The positive cells are the marginal cells in the stria vascularis, and the hair cells in Corti's organ and the saccule. However, under morphological analysis by light microscope, these cells lack the features characteristic of apoptosis. Moreover, no cells staining positive for Bcl-2 and Bax are found in any structures of the inner ears. These results suggest that positive TUNEL staining in this model does not indicate apoptosis and apoptosis may be not involved in autoimmune inner ear disease. PMID- 12624502 TI - Characteristics of fricatives and sentence duration after cochlear implantation. AB - Ten postlingually deafened patients (5 male, 5 female) were examined after cochlear implantation to measure improvements in their quality of speech. Parameters such as the spectral maximum of fricatives and the duration of utterances were analysed in speech recordings taken at regular intervals after implantation. The speech samples were recorded in an audiological chamber. Parameters were analysed using ST(x) (S-Tools Software). Frequency analyses based on the fast Fourier transform and spectral estimation methods, as well as fundamental frequency and formant extraction (cepstrum, LPC = linear prediction coding) and digital filter implementations were prepared. The results indicate a tendency towards improvement in the spectral maximum of the fricatives and affricates and a shortening of the duration of the fricative parts in affricates and of sentences in nearly all our subjects. These results showed the restored auditory feedback produced by cochlear implantation to have a favourable effect on speech production. PMID- 12624503 TI - Expression of p16 protein and hypermethylation status of its promoter gene in adenoid cystic carcinoma of the head and neck. AB - Reduced expression of the p16 gene product (protein), an inhibitor of cyclin-D dependent protein kinase which regulates cell cycle at the G1/S boundary, is implicated in tumor progression in various neoplasms. Hypermethylation of the p16 promoter gene has recently been suggested to be one of the reasons for the reduced protein expression. To explore the role of p16 in the biological behavior of adenoid cystic carcinomas (ACC), we investigated the immunohistochemical expression of p16 protein in 38 ACC tumors (32 primary, 3 recurrent, and 3 metastatic tumors) and the methylation status of its promoter gene. We also examined their relationships to the histological grade of malignancy. Positive reaction of p16 protein was demonstrated in the nuclei of luminar cuboidal cells in areas with tubular patterns. The reactions were reduced in the areas with solid or large cribriform patterns. The levels of p16 expression correlated with the histological grade of malignancy. Recurrent or metastatic tumors did not differ with respect to histological grades from the original tumor except for 1 case, in which p16 expression was reduced compared to the primary tumor. Methylation-specific PCR demonstrated the hypermethylation status of the p16 promoter gene in 4 of 22 primary tumors (21%), all of which showed negative or low expression of the p16 protein. The study indicated that p16 expression was reduced in ACC cases of higher histological grade of malignancy and that hypermethylation of its promoter gene may be involved in its process in some cases. PMID- 12624504 TI - Contribution of virtual endoscopy to the investigation of laryngotracheal pathological conditions. AB - Three-dimensional CT scan imaging obtained by using helicoidal CT scan provides the basis for an endoscopic exam said to be virtual since no invasive procedure is actually performed. Compared to optical endoscopy this easily accessible exam offers additional information especially for the analysis of the infraglottic and tracheal areas, which are two anatomically rigid segments. This property facilitates their three-dimensional reconstruction. Our study encompassed 6 patients presenting with a stenosis of the laryngotracheal tract. In 5 of them it was possible to correlate optical and virtual endoscopic imaging. Coupling both exams significantly improved the diagnostic investigation and facilitated the management of the disease. However, the real contribution of virtual endoscopy to the exploration of tumoral conditions still remains to be determined given the low degree of tissue resolution. As a consequence parietal and extraparietal lesional spreading is more accurately assessed by axial scan imaging. PMID- 12624505 TI - Cochlear implantation in Mondini dysplasia. AB - The use of cochlear implantation to treat patients with inner ear malformations such as Mondini dysplasia has been increasingly successful. Until now, conventional hearing aids in these patients have not performed well. Consequently, the hearing problem for patients with this condition has been somewhat improved with the use of cochlear implants. Various results of cochlear implantation have been reported in these patients so far. This is a report of 5 patients with Mondini malformation who have undergone cochlear implant surgery. PMID- 12624506 TI - Familial isolated unilateral large vestibular aqueduct syndrome. AB - Large vestibular aqueduct syndrome (LVAS) is one of the most frequent inner ear malformations. It is associated with varying degrees of hearing loss, that may be progressive, sudden or fluctuant; nevertheless normal hearing in cases with LVAS has been described. Although unilateral LVAS has been described, all patients show clinical and radiological findings in both ears, in contrast to the family presented here, where 2 siblings are affected by unilateral sensorineural hearing loss associated with ipsilateral LVAS, with an autosomal recessive pattern. In order to search for the origin of this inner ear malformation, mutations in genes GJB2 (connexin-26 gene), PDS (Pendred's syndrome gene) and mitochondrial 12sRNA were studied, without positive results. In this paper we hypothesize the probable embryological origin for this malformation. PMID- 12624507 TI - Pneumocystis carinii infection of the middle ear and external auditory canal. Report of a case and review of the literature. AB - We present a 50-year-old male with Pneumocystis carinii infection involving the middle ear and the external auditory canal as the first manifestation of a previously unknown HIV infection. In case of therapy-resistant otitis with a polypoid mass in the external auditory canal histological evaluation should be considered to rule out malignancy or pathogens, like Pneumocystis carinii, that cannot be cultured. Oral or intravenous antiprotozoal agents are the treatment in line with current practice. Exploration of the os petrosum is never required. PMID- 12624508 TI - Malignant ectomesenchymoma of the nose. Case report and review of the literature. AB - To the best of our knowledge, a malignant ectomesenchymoma of the nose has not yet been reported. This rare tumour is composed of a mesenchymal element--usually a rhabdomyosarcoma--and a neuroectodermal element, often a neuroblastoma or a malignant schwannoma. Most malignant ectomesenchymomas have been found in children (mainly boys), but 5 adult cases, including our case, have also been reported. A 62-year-old man presented with a nasal obstruction on the right side of the nose. Diagnostic nasal endoscopy revealed a mass in the dorsal part of the right nasal cavity, which was treated by endoscopic surgery. The postoperative course was uneventful, and subsequent radiotherapy well tolerated. Since rhabdomyosarcoma is the most common component of this neoplasm, the treatment protocol for rhabdomyosarcoma would have appeared appropriate. This report reviews the literature, summarises the pathological features and possible histogenesis of the tumour, and outlines recommended treatments for malignant ectomesenchymoma. PMID- 12624509 TI - Unexpected expansive paranasal sinus mucormycosis. AB - Mucormycosis infection of the paranasal sinuses is a very rare but serious condition. Patients are usually immunocompromised by systemic disease. We report an unexpected case of expansive paranasal sinus mucormycosis in an immunocompetent European treated successfully with surgical debridement and intravenous amphotericin B. The pathogenesis, diagnosis and treatment of paranasal sinus mucormycosis is discussed. PMID- 12624510 TI - New interdisciplinary approach for removing large clivus and upper cervical spine tumors. AB - We have developed a new interdisciplinary approach for removing large clivus and upper cervical spine tumors. This approach is a combination of the Le Fort I osteotomy, midfacial degloving, and median labiomandibular glossotomy. Our approach gives an excellent, wide surgical field from the nasopharynx, including the base of the skull, to the base of the tongue and permits sufficiently safe extirpation of clivus and upper cervical spine tumors that may not be removed by transoral or transcervical approaches. Our approach not only incorporates the merits of each approach but also creates a larger surgical field that may be modified or expanded to accommodate the removal of more bulky tumors in this region. This novel approach will facilitate more successful resection of tumors arising between the nasopharynx, including the skull base, and the retropharyngeal area. PMID- 12624511 TI - Feasibility of sentinel lymph node radiolocalization in neck node-negative oral squamous cell carcinoma patients. AB - We examined the reliability of the use of lymphoscintigraphy (LS) and a hand-held gamma probe (GP) to identify the sentinel lymph node (SLN), and sequentially determined the feasibility of SLN radiolocalization in clinical neck node negative oral squamous cell carcinoma (SCC) patients. A radiolabel with the unfiltered (99m)Tc tin colloid was injected submucosally around the primary site followed by LS at 2-hour intervals. Preoperative localization was performed with GP. After en bloc removal of the regional lymphatics, histopathologic results for the nodes were compared with the SLN radiolocalization. The LS and GP counts were well correlated, and there was concordance between SLN and neck node status in 7 of 8 cases (87.5%). We thus considered that our concept in this study was valid in determining the necessity for neck dissection for those node-negative oral SCC patients. PMID- 12624512 TI - Patterns of failure following radiation with and without chemotherapy in patients with nasopharyngeal carcinoma. AB - PURPOSE: Aim of this study was to retrospectively evaluate patterns of failure, results, and prognostic factors for patients with nasopharyngeal cancer (NPC) following radiotherapy (RT) with and without concurrent chemotherapy (RCT). PATIENTS AND METHODS: Between 1978 and 1999, a total of 101 patients with NPC were treated in our hospital, of whom 53 received external megavoltage RT alone with a median total dose of 76 Gy (1978-1988), and 48 patients had RCT (1989 1999). For RCT a combination of 5-FU and cisplatin was used together with a median total dose of 72 Gy. Patterns of relapse, survival rates and toxicity as well as prognostic factors were evaluated retrospectively. RESULTS: RCT was associated with a marked reduction in distant metastases: 6/48 (13%) vs. 17/53 (32%) after RT alone. Locoregional tumor persistence was only marginally lower with RCT: 10/48 (21%) vs. 17/53 (32%) following RT. Patients with RCT demonstrated a survival advantage compared to those with RT alone (5-year overall survival (OS): 64% vs. 44%, p = 0.1). OS, disease-specific survival and locoregional control rates were 53, 57, and 78% at 5 years and 47, 51 and 78% at 10 years, respectively. OS was significantly affected by histology (p = 0.007), the patients' age (p = 0.009) and gender (p = 0.01). CONCLUSION: This retrospective study provides further evidence that both reduction of distant metastasis and enhanced local tumor control by combined radiochemotherapy may be associated with improved survival rates in NPC compared to radiation alone. Concurrent RCT is therefore considered the preferable treatment option, however, confirmation in randomized trials is still warranted. PMID- 12624513 TI - Short-time splenic irradiation for splenomegaly. AB - BACKGROUND: Splenic irradiation is routinely used in the supportive treatment of lymphoepithelial and hemopoietic diseases associated with splenomegaly. A new short-time irradiation schedule with conventional dosage (Group A: 2 Gy/fract.) was compared retrospectively with low-dose prolonged treatment schedules (Group B: < or = 1 Gy/fract.) to establish its tolerance and its efficacy in terms of relieving splenomegaly-associated symptoms. PATIENTS AND METHOD: Between 12/1996 and 3/2002 49 patients (6 with CLL, 14 with CML, 6 with NHL, 16 with MPD, 6 with OMF and 1 with AML) underwent 85 treatment courses (13 courses low-dose prolonged treatment and 72 courses short-time treatment).The spleens had been exposed to pretreatment doses of 3-70 Gy (mean 20.75 Gy). Splenic size was 12-35 cm (mean 22.76 cm). One treatment was done with 60Co. 9-20 MeV electron beams were used in 23 treatments and 6 MV photons in 61 treatments. Blood counts were monitored daily. Clinical improvement (pain relief, improvement of splenomegaly-associated symptoms), spleen volume reduction and effects on blood counts were evaluated and documented at the end of the treatment. RESULTS: Of the 85 treated patients, 55 (Group A: 47/72, Group B: 8/13) showed a > 10% reduction of spleen volume and 62 (Group A: 57/72, Group B: 5/13) clinical improvement. 15 of 49 patients underwent multiple irradiation (2-8 courses) without clinical or hematologic complications. The intervals between the courses were 2-19 months. Rapid response in terms of reduction in splenic size in 23 patients permitted field reduction during treatment. Significant hematologic abnormalities were absent throughout. CONCLUSION: The short-time treatment schedule ensured a rapid response with relief of pain and improvement of gastrointestinal and pulmonary symptoms at an overall treatment time shorter than that of low-dose prolonged treatment schedules. No differences in terms of hematologic abnormalities were found between single doses of 2 Gy and < or = 1 Gy. PMID- 12624514 TI - Breast cancer: monitoring response to neoadjuvant chemotherapy using Tc-99m sestamibi scintimammography. AB - BACKGROUND: Aim of the study was to assess the value of scintimammography using Tc-99m sestamibi in the evaluation of tumor response to neoadjuvant chemotherapy. MATERIAL ANS METHODS: Results were calculated for 9 patients undergoing neoadjuvant chemotherapy. Scintimammography using 740 MBq Tc-99m sestamibi was performed before, during and after chemotherapy, and sestamibi uptake was scored visually and semiquantitatively to evaluate tumor response. RESULTS: In the case of complete response (n = 3) sestamibi uptake decreased 8 days after beginning neoadjuvant chemotherapy and normalized in the following course. Focal uptake decreased more slowly in patients with partial response (n = 3), who showed clear, persisting tracer accumulation after therapy. The patients without response (n = 3) showed a persisting high tumor activity even after chemotherapy was completed. CONCLUSIONS: The preliminary data suggest that in contrast to other imaging modalities scintimammography appears to yield early information regarding tumor response to neoadjuvant chemotherapy. PMID- 12624515 TI - Development of a flow cytometric method to determine DNA ploidy of oesophageal cancer cells obtained by forceps biopsy samples during oesophago-gastro duodenoscopy. AB - BACKGROUND: The DNA content of oesophageal tumour cells is a prognostic factor in untreated patients. To investigate whether DNA ploidy is useful to select patients for neoadjuvant therapy it is of interest to develop a method allowing reliable flow cytometric analysis of the DNA content of tumour cells obtained by forceps biopsy during endoscopy before start of therapy. METHODS: Freshly frozen forceps biopsy samples from 30 patients with oesophageal cancer were disaggregated. DNA was stained with propidium iodide and ploidy was determined by flow cytometry. To enhance sensitivity epithelial cells were simultaneously labelled with anti-cytokeratin antibodies. Results were compared with image analysis. To evaluate the sampling error, parallel measurements were done in 10 patients by image analysis on forceps biopsies obtained during endoscopy before surgery and on the resected tumour. RESULTS: The sensitivity to detect aneuploidy was lower for standard flow cytometry than for image analysis (13 versus 33%). The overall sensitivities were identical using a double labelling technique with additional cytokeratin-staining of the epithelial cells, but divergent results were obtained in 2 cases, where detection of aneuploidy was either possible with image analysis or with double labelling flow cytometry only. DNA content of samples gained by forceps biopsies and surgically resected tumours was concordant in 8 of 10 cases. In 2 patients, aneuploidy was detected only in the surgically resected tumour but not in the pre-operatively obtained forceps biopsies. CONCLUSIONS: A flow cytometric method for routine determination of the DNA ploidy of cells obtained by forceps biopsies from patients with oesophageal cancer was developed and evaluated against image analysis. The technique allows the prediction of DNA content before tumour resection, and might be used for optimising therapy and the patient's quality of live. PMID- 12624516 TI - Quality of life (QoL) as predictive mediator variable for survival in patients with intracerebral neoplasma during radiotherapy. AB - BACKGROUND: The prognosis for patients with malignant astrocytoma or brain metastases is often fatal despite intensive therapy. Therefore we wished to elucidate whether the quality of life (QoL) is a determinant of overall survival (OAS). PATIENTS AND METHODS: From 1997 to 2000 153 patients with brain tumours were screened; 39 patients (26%) refused to participate and further 47 patients were excluded (cerebral impairment 14%, amaurosis/ language problems 3%, Karnofsky performance score < 50% 7%, death 8%, non-compliance 7%). Thus, 57 patients were analysed (33 with primary brain tumours, 24 with brain metastases). With the FACT-G questionnaire cancer-specific aspects of health-related QoL were assessed. RESULTS: Patients with metastases showed a lower QoL in the physical sphere than patients with astrocytoma, but there were no significant differences in OAS. Median survival of patients with good QoL was 31.3 months versus 14.2 months in patients with bad QoL. Only the two variables 'living with a spouse' and FACT-G sum score had a statistically significant influence on survival (p = 0.033 and p = 0.003) modelled by the Cox-PH regression. Patients who did not live with a spouse had shorter survival times than the other patients. CONCLUSION: Health-related QoL can serve to identify a patient group with higher risks of death. PMID- 12624517 TI - Stromal antigen targeting by a humanised monoclonal antibody: an early phase II trial of sibrotuzumab in patients with metastatic colorectal cancer. AB - BACKGROUND: A novel immunological approach to colon cancer therapy is the antibody targeting of the fibroblast activation protein (FAP), which is highly expressed by stroma cells of this tumour. Unconjugated sibrotuzumab (BIBH 1), which is a humanised version of the murine anti-FAP mAb F19, was investigated for its anti-tumour activity, safety and pharmacokinetics. PATIENTS AND METHODS: Patients with metastatic colorectal cancer received weekly intravenous infusions of unconjugated sibrotuzumab at a dose of 100 mg over 12 scheduled weeks. The study was implemented as an open-label, uncontrolled, multicentre trial. RESULTS: 25 patients were enrolled. Patients had one or more measurable lesions, predominantly liver lesions, at baseline. At least 8 repeated weekly infusions of sibrotuzumab in 17 evaluable patients did not result in complete or partial remission. Rather, ongoing tumour progression was noted in all patients except for 2 patients with stable disease. However, progressive disease was also observed post-study in these 2 patients who received 1 and 6 additional infusions, respectively, of sibrotuzumab. Sibrotuzumab exhibited 2-compartment pharmacokinetics with a dominant terminal phase and t1/2 beta = 5.3 +/- 2.3 days. Adverse drug reactions (rigors/chills, nausea, flushing and one incidence of bronchospasm) were observed in 5 patients. Of the 24 patients given 2 or more infusions of sibrotuzumab, antibodies against sibrotuzumab were found in 3 patients (12.5%) after 4-12 infusions. CONCLUSIONS: Sibrotuzumab was well tolerated and safe. The minimal requirement for the continuation of this exploratory trial, of at least one complete or partial remission, or equivalently, of 4 patients with stable disease, was not met. PMID- 12624518 TI - Cancer-related fatigue in patients attending oncological rehabilitation programs: prevalence, patterns and predictors. AB - BACKGROUND: Fatigue is one of the most frequent symptoms in cancer patients. It is the aim of medical and psychosocial rehabilitation programs to reduce physical impairments and psychosocial distress. There are no reliable data on the prevalence or the pattern of fatigue in patients attending such rehabilitation programs. We therefore initiated a cross-sectional study to evaluate this phenomenon. PATIENTS AND METHODS: Using the EORTC QLQ-C30, HADS and MFI questionnaires in 144 patients with different malignant diseases the incidence, patterns and necessity of treatment of fatigue were evaluated. RESULTS: One third of the patients were diagnosed with breast cancer, 27% with gastrointestinal cancers and 20% suffered from haematologic/lymphatic diseases. There was a good correlation between fatigue levels in the 3 instruments used. 22% of all patients showed significant symptoms of fatigue. The univariate analysis did not reflect any significant differences in severity of fatigue with respect to diagnosis, haemoglobin levels, medical and therapeutic history or sociodemographic data. However, using cluster analysis we could identify 3 groups of patients that differ significantly in fatigue and quality-of-life measurements. CONCLUSION: We conclude from our data that fatigue is an important phenomenon in cancer patients who attend medical and psychosocial rehabilitation programs. This has an impact on the ability and degree to reintegrate these patients into everyday life. PMID- 12624519 TI - Uterine cervical metastasis of breast cancer: a rare complication that may be overlooked. AB - BACKGROUND: Metastasis of distant malignancies to the cervix uteri is a rare occurrence and the frequency is approximately 4% for all tumours. However, the frequency of cervical metastasis of breast cancer is much lower and is estimated to range between 0.8 and 1.7%. With the exception of ovarian metastases, secondary tumours of the female genital tract are rather uncommon. Therefore, these conditions pose diagnostic problems for the clinician. PATIENT: A 40-year old woman with the diagnosis of invasive ductal cell carcinoma of the right breast underwent mastectomy with dissection of axillary lymph nodes in 1998. Subsequently, the patient received 6 cycles of chemotherapy with cyclophosphamide, methotrexate and fluorouracil. The initial tumour stage was pT2, pN0 (0/13), M0, G2. The oestrogen and progesterone receptors were positive and expression of the C-erb-B2 coding oncogene was negative. Gynaecological and ultrasonographic examination revealed a normal cervix without evident lesions. Exfoliative cytology was negative. 14 months after treatment the patient presented with an axillary relapse and surgery, second-line chemotherapy with doxorubicine and radiation therapy of the chest wall and the axilla were performed. The patient developed liver metastases 14 months later and at this time ultrasonographic pelvic examination revealed a 2.2 cm tumour of the cervix with good vascularisation. The patient had no clinical symptoms, i.e. no vaginal bleeding or discharge. Sonomorphologically this tumour appeared as a leiomyoma of the cervix. Cervical biopsies and curettage, however, revealed metastatic carcinoma expressing oestrogen and progesterone receptors consistent with the primary breast cancer. Under palliative chemotherapy with docetaxel progression of liver metastases and cervical metastasis occurred and the patient died 9 months later. CONCLUSION: Metastatic involvement of the cervix should be considered in women with a history of breast cancer who present with vaginal bleeding or suspicious changes of the cervix on transvaginal ultrasound. Therefore, gynaecological and ultrasonographic examination of the pelvis represent an important part of the follow- up investigations in women with primary breast cancer. PMID- 12624520 TI - Recurrent gastric adenocarcinoma with unusual metastatic localization and excellent response to docetaxel and 5-FU continuous infusion. AB - BACKGROUND: Metastatic gastric cancer is usually treated with cisplatin- and 5-FU based chemotherapy regimens. There are good data for the combination regimen ECF (epirubicin, cisplatin and 5-FU), which is therefore often regarded as a reference treatment. Docetaxel shows promising activity against gastric cancer as single agent and in combinations. To develop a well-tolerable combination chemotherapy for an ambulant setting we initiated a randomized phase II study, comparing docetaxel and 5-FU continuous infusion (DF) with ECF. CASE REPORT: A 66 year-old patient with the history of a curatively resected gastric cancer 2 years previously presented with abdominal masses and lesions in his spleen. Histology proved metastases of gastric adenocarcinoma. The patient was treated with docetaxel (75 mg/m2, d1) and 5-FU continuous infusion (200 mg/m2/d, d1-21, q3w) within our study. Already after 2 cycles of chemotherapy he showed symptomatic improvement and partial remission of his tumor, which was confirmed after the 3rd cycle. In our ongoing study so far 50 patients are evaluable for response. Objective tumor response (CR + PR) could be documented in 44% of patients in the DF arm as well as in the ECF arm. CONCLUSION: Docetaxel and 5-FU continuous infusion is an active regimen which could possibly be used as an alternative to established treatment protocols. PMID- 12624521 TI - Evaluation of posttraumatic psychological problems in cancer patients. AB - The introduction of the diagnosis 'posttraumatic stress disorder' (PTSD) in cancer patients in the diagnostic nomenclature and the application of assessment to investigate prevalence, risk factors and comorbidity are discussed. The diagnosis PTSD in cancer patients makes it necessary to modify the trauma criteria. Data to identify patients at risk for developing PTSD in the group of cancer patients are incomplete. Prospective, longitudinal studies are needed to define the characteristics and course of PTSD in the cancer setting. With these data it would be possible to identify the population at risk and the best time for assessment and therapeutic intervention. PMID- 12624522 TI - Granulocyte transfusion therapy for treatment of infections after cytotoxic chemotherapy. AB - Opportunistic fungal infections and antibiotic-refractory bacterial infections remain important causes of morbidity and mortality in neutropenic individuals. Furthermore, the expanding use of dose-intensive cancer treatment strategies has increased the frequency of prolonged neutropenia. Therefore, the transfusion of granulocytes should be a logical therapeutic approach. Substantial progress has been made in the field of granulocyte transfusion therapy during the past decade. Interest in granulocyte transfusion therapy has been rekindled by both the use of hematopoietic growth factors to mobilize neutrophils and modern leukapheresis techniques. Moreover, promising results were observed in the use of community donors and in granulocyte storage experiments, which could enhance the ability of blood banks for institution of granulocyte concentrates. Recent clinical trials suggest that granulocyte transfusion therapy may be effective and well-tolerated in the neutropenic patient affected by life-threatening infections. These results must be confirmed in controlled, clinical trials. PMID- 12624523 TI - Use of erythropoietin in patients with multiple myeloma. AB - The prevalence of tumour anaemia in patients with multiple myeloma is greater than 80%. At the time of diagnosis 20% of these patients are already anaemic. In about 70% of patients with multiple myeloma, recombinant human erythropoietin (r HuEPO) leads to a reduction in transfusion frequency, resulting in a drop in transfusion- related side-effects like infections and immune reactions, iron overload and hyperviscosity which often negatively influence the course of disease. A further reason for the use of erythropoietin is to achieve and maintain high haemoglobin levels (11-12 g/dl), which are of considerable prognostic significance in patients with multiple myeloma. Increasing Hb levels with r-HuEPO also improve the quality of life of patients, thus leading to better therapy compliance. The trade-off between high costs of an erythropoietin treatment and lower indirect costs (infusion material, personal equipment, patient transport costs, etc.) should be evaluated. Nevertheless, an exact definition of patients for whom the use of erythropoietin is beneficial is warranted. The pathogenesis of anaemia and the clinical experiences of erythropoietin in patients with multiple myeloma are discussed. PMID- 12624524 TI - G protein selectivity is regulated by multiple intracellular regions of GPCRs. AB - GTP-binding protein coupled receptors (GPCRs) bind to a vast diversity of extracellular ligands to regulate a wide variety of physiological responses. Upon binding of extracellular ligands, these seven-transmembrane-spanning receptor molecules couple to one or several subtypes of G protein which reside at the intracellular side of the plasma membrane to trigger intracellular signaling events. Amid the large structural diversity at the intracellular regions of GPCRs, there are only 18 different subtypes of G protein belonging to four subfamilies. The question of how GPCRs select and activate a single or multiple G protein subtype(s) has been the topic of intense investigations. This review will attempt to summarize the available data on the structural determinants in GPCRs that regulate the selectivity of G protein activation. The available data suggest that G protein can be activated by structurally diverse cationic alpha-helical structures with no obvious homology in primary sequence. The selectivity of receptor-G protein coupling is maintained by a combination of two functional domains at the intracellular region. One is the 'activation domain' which can activate multiple G protein subtypes, while the other is the 'selectivity domain' which restricts the coupling to the desired signaling pathway(s). A slight change in the conformation at these two functional domains can affect the fidelity of G protein selectivity. This hypothesis can account for the vast structural diversity of GPCRs which link a fascinating variety of extracellular inputs, yet couple to a limited number of intracellular signaling pathways. PMID- 12624525 TI - Roles of SNARE proteins and synaptotagmin I in synaptic transmission: studies at the Drosophila neuromuscular synapse. AB - The roles of SNARE proteins, i.e. neuronal Synaptobrevin (n-Syb), SNAP-25 and Syntaxin 1A (Syx 1A), and Synaptotagmin I (Syt I) in synaptic transmission have been studied in situ using mutant embryos or larvae that lack these molecules or have alterations in them. Because of the ease of genetic manipulation, the Drosophila neuromuscular synapse is widely used for these studies. The functional properties of synaptic transmission have been studied in mutant embryos using the patch-clamp technique, and in larvae by recording with microelectrodes. A major vesicular membrane protein, n-Syb, is indispensable for nerve-evoked synaptic transmission. Spontaneous synaptic currents (minis), however, are present even in embryos totally lacking n-Syb (N-SYB). Furthermore, Ca(2+)-independent enhancement of mini frequency induced by hypertonic sucrose solutions (hypertonicity response) is totally absent in N-SYB. Embryos that have defects in SNAP-25 (SNAP-25) have similar but milder phenotypes than N-SYB. The phenotype in synaptic transmission was most severe in the synapse lacking Syx 1A. Neither nerve-evoked synaptic currents nor minis occur in embryos lacking Syx 1A (SYX 1A). No hypertonicity response was observed in them. Syt I binds Ca(2+) in vitro and probably serves as a Ca(2+) sensor for nerve-evoked synaptic transmission, since nerve-evoked synaptic currents were greatly reduced in embryos lacking Syt I (SYT I). Also, Syt I has a role in vesicle recycling. Interestingly, the Ca(2+) independent hypertonicity response is also greatly reduced in SYT I. Minis persist in mutant embryos lacking any of these proteins (n-Syb, SNAP-25 and Syt I), except Syx, suggesting that minis have a distinct fusion mechanism from that for fast and synchronized release. It appears that these SNARE proteins and Syt I are coordinated for fast vesicle fusion. Minis, on the other hand, do not require SNARE complex nor Syt I, but Syx is absolutely required for vesicle fusion. The SNARE complex and Syt I are indispensable for the hypertonicity response. None of these molecules seem to serve for selective docking of synaptic vesicles to the release site. For further studies on synaptic transmission, the Drosophila neuromuscular synapse will continue to be a useful model. PMID- 12624526 TI - N-methyl-D-aspartate receptor antagonist activity in traditional Chinese stroke medicines. AB - Traditional Chinese medicine (TCM) has a long history in stroke therapy and its therapeutic efficacy has been confirmed by clinical studies. The molecular basis of the neuroprotective effects is unknown. We wondered whether or not the neuroprotective effect of TCMs might be due to their N-methyl-D-aspartate (NMDA) receptor (NMDAR) antagonist properties. We used the patch-clamp technique to screen 22 TCM stroke drugs for NMDAR antagonist activity in cultured cortical neurons. The drugs were also screened for their ability to abate NMDA-induced neurotoxicity. Aqueous extracts of Scutellaria baicalensis, Stephania tetrandra, and Salvia miltiorrhiza blocked currents induced by NMDA (200 microM, 10 microM glycine, 0 Mg2+) at a holding potential of -80 mV by 83.45+/-4.34, 38.65+/-7.50, and 52.97+/-1.78%, respectively. The block of the NMDA-evoked currents was voltage-dependent and showed a negative slope conductance reminiscent of Mg2+. Atomic absorption spectrophotometry revealed the presence of 12.5, 2, and 8.7 mM Mg2+ in the extracts of S. baicalensis,S. tetrandra, and S. miltiorrhiza, respectively. None of these extracts blocked NMDA-induced neuronal death. The Uncaria rhynchophylla extract blocked NMDA-evoked currents by 54.98+/-8.61% even at +60 mV and reduced NMDA-induced neuronal death by 59.13+/-3.52%. NMDAR antagonist activity may underlie the neuroprotective effects of this TCM. Some TCM drugs may exert therapeutic effects due to their Mg2+ content. PMID- 12624527 TI - The stimulatory effect of cannabinoids on calcium uptake is mediated by Gs GTP binding proteins and cAMP formation. AB - Cannabinoids are neurodepressive drugs that convey their cellular action through G(i/o) GTP-binding proteins which reduce cAMP formation and Ca(2+) influx. However, a growing body of evidence indicates that the stimulatory effects of cannabinoids include the elevation in cAMP and cytosolic Ca(2+) concentration. The present study expands our previous findings and demonstrates that, in N18TG2 neuroblastoma cells, the cannabinoid agonist desacetyllevonantradol (DALN) stimulates both cAMP formation and Ca(2+) uptake. The stimulatory effect of DALN on cAMP formation was not eliminated by blocking Ca(2+) entry to the cells, while its stimulatory effect on Ca(2+) uptake was abolished by blocking cAMP-dependent protein kinase. Furthermore, elevating cAMP by forskolin stimulated calcium uptake, while elevating the intracellular Ca(2+) concentration by ionomycin or KCl failed to stimulate cAMP formation. These findings suggest that cAMP production precedes the influx of Ca(2+) in the cannabinoid stimulatory cascade. The stimulatory effect of DALN on calcium uptake resisted pertussis toxin treatment, and was completely blocked by introducing anti-G(s) antibodies into the cells, indicating that the stimulatory activity of cannabinoids is mediated by G(s) GTP-binding proteins. The relevance of the cellular stimulatory activity of DALN to the pharmacological profile of cannabinoid drugs is discussed. PMID- 12624528 TI - Effect of protein phosphorylation on neurite outgrowth in cultured embryonic Xenopus spinal neurons. AB - Intracellular signaling pathways involved in neurite outgrowth have been extensively studied in a variety of cell systems. While most of these studies utilized continuous neuronal-like cell lines, fewer studies have been conducted in primary neuronal culture. One primary culture system that has recently been used to dissect the signaling pathways involved in axon guidance consists of spinal neurons derived from embryonic Xenopus laevis. In this study, we used Xenopus to study neurite outgrowth by treating neuronal cultures with pharmacological agents that activate or inhibit various protein kinases or that inhibit protein phosphatases. We found that agents which affected signaling via cAMP-dependent protein kinase, calmodulin, cyclin-dependent kinase 5, or protein phosphatases had effects on Xenopus neurite outgrowth that were similar to those reported in other primary neurons or in neuronal-like cell lines. However, agents which affected protein kinase C signaling had effects on Xenopus neurite outgrowth that were distinct from those reported in neuronal-like cell lines. Although continuous cell lines have several advantages for the dissection of signaling pathways involved in neurodevelopment, these observations underscore the importance of also using primary neurons to examine these pathways. PMID- 12624529 TI - Role of ghrelin in the control of growth hormone secretion in prepubertal rats: interactions with excitatory amino acids. AB - Ghrelin is a 28-amino-acid peptide, with an essential n-octanoyl modification at Ser3, that elicits growth-hormone (GH) secretion in rats and humans. At present, the mechanisms of ghrelin action and its interactions with other systems controlling GH secretion remain poorly characterized. In this context, the present study was undertaken to obtain information about ontogeny and possible gender differences in the GH-releasing activity of ghrelin, and to delineate its primary site(s) of action at the hypothalamus and/or pituitary. In addition, the interactions between ghrelin and other relevant signals in the control of GH secretion, such as excitatory amino acids (EAAs), nitric oxide (NO) and serotonin, were assessed. Experiments were carried out in infantile-prepubertal animals, when GH pulsatility is not yet established. Systemic administration of ghrelin (25 nmol/rat, i.p.) to 5-, 10- and 23-day-old male and female rats increased plasma GH levels from day 10 onwards. This action was NO dependent, since it disappeared in 23-day-old males after pretreatment with an inhibitor of NO synthase (NAME). Similarly, central infusion of ghrelin (3 nmol/rat, i.c.v.) elicited GH responses in 10- and 23-day-old animals significantly higher than after systemic administration. By contrast, in vitro challenge of pituitary tissue with increasing doses of ghrelin (10(-9)-10(-7) M) failed to enhance GH release into the incubation medium, whereas stimulation with GH-releasing hormone (GHRH; 10(-7) M) or GHRP-6 (10(-7) M) was effective. Finally, effects of ghrelin were blocked by pretreatment with MK-801 and NBQX antagonists of EAA ionotropic receptors and after manipulation of endogenous serotoninergic tone. In addition, the potent releasing activity of EAA agonists NMDA and AMPA was blunted by pretreatment with D-Lys3-GHRP-6, a selective antagonist of the cognate ghrelin receptor, i.e. the GH-secretagogue receptor. In conclusion, our results demonstrate that GH-releasing activity of ghrelin appears early in the infantile period, is NO dependent and involves a primary hypothalamic site of action. The data also demonstrate for the first time the existence of a cross-talk between ghrelin and other neurotransmitter systems, such as EAAs and serotonin, in precise control of GH secretion. PMID- 12624530 TI - Ontogeny and tissue-specific regulation of ghrelin mRNA expression suggest that ghrelin is primarily involved in the control of extraendocrine functions in the rat. AB - Ghrelin is a 28-amino-acid gastric peptide that potently stimulates growth hormone (GH) secretion in vivo and in vitro. Ghrelin-expressing cells have been found in the oxyntic region of the stomach and in the arcuate nucleus of the hypothalamus. The aim of this work was to investigate the regional distribution and developmental changes in ghrelin mRNA levels in the pituitary, hypothalamus and gastrointestinal (GI) tract of the rat using a semiquantitative RT-PCR assay. We also describe the effects of ghrelin immunoneutralization in late gestation and those resulting from induction of an isolated GH deficiency in adult rats. Ghrelin mRNA was already expressed in the fetus by embryonic day 12 (E12), by E17 most of ghrelin mRNA was in the trunk. At E17, in situ hybridization did not reveal a clear expression of ghrelin mRNA in fetal stomach but showed high ghrelin mRNA levels in the placenta. In the pituitary gland, levels of ghrelin mRNA were high after birth but declined significantly with puberty, whereas in the hypothalamus they were barely detectable at birth and remained very low at all subsequent time points tested. In the GI tract, ghrelin mRNA levels were high from birth to 270 days of life. Immunoneutralization of ghrelin at E16 had no effect on survival or development. Rats showed normal somatotropic function, ghrelin expression and onset of puberty. In young adult rats, passive immunization against GHRH did not affect ghrelin mRNA levels in the pituitary, hypothalamus and stomach. Only a 72-hour fasting period induced a significant increase in ghrelin mRNA levels in the stomach, but not in the pituitary and hypothalamus. These results strongly indicate that ghrelin is an important GI hormone expressed early in life and primarily sensitive to nutritional status. PMID- 12624531 TI - Distribution of fluorescence following injection of recombinant adeno-associated virus encoding green fluorescent protein into the paraventricular nucleus. AB - We have used recombinant type 2 adeno-associated virus to deliver the gene encoding green fluorescent protein into the central nervous system of adult rats. Gene expression, determined by fluorescent microscopy, was observed not only at the site of injection but also in axons following known neuroanatomical pathways. We have demonstrated a spread of enhanced green fluorescent protein from the paraventricular nucleus of the hypothalamus into the median eminence and neurohypophysis. Cell bodies containing enhanced green fluorescent protein were also visualized in the paraventricular nucleus contralateral to the side of injection. These findings suggest that gene transfer by recombinant adeno associated virus could be used as a tool to investigate hypothalamic-pituitary interactions and, elsewhere in the central nervous system, to trace axonal pathways. PMID- 12624532 TI - Expression of D1 and D2 dopamine receptors in the hypothalamus and pituitary during the turkey reproductive cycle: colocalization with vasoactive intestinal peptide. AB - The regulation of avian prolactin (PRL) secretion and PRL gene expression is influenced by hypothalamic vasoactive intestinal peptide (VIP), the PRL-releasing factor in avian species. Recent evidence indicates that D(1) and D(2) dopamine (DA) receptors play a pivotal role in VIP and PRL secretion. The differential expression of DA receptors located on hypothalamic VIP neurons and anterior pituitary cells may affect the degree of prolactinemia observed during the turkey reproductive cycle. The relative expression of D(1D) and D(2) DA receptor subtype mRNA was quantitated using in situ hybridization histochemistry (ISH). D(1D) and D(2) DA receptor mRNA was found expressed throughout the hypothalamus and pituitary. The expression of D(1D) DA receptor mRNA in the hypothalamus was found to be 6.8-fold greater than that of D(2) DA receptor mRNA. Higher D(1D) DA receptor mRNA content was found in the anterior hypothalamus (3.6-fold), the ventromedial nucleus (2.0-fold), the infundibular nuclear complex (INF; 1.9 fold), and the medial preoptic nucleus (1.5-fold) of laying hens as compared to that of reproductively quiescent non-photostimulated hens. The levels seen in incubating hyperprolactinemic hens were essentially the same as in laying hens, except for the INF where levels were 52% higher. During the photorefractory stage (hypoprolactinemia), the D(1D) DA receptor mRNA was at its lowest level in all areas tested. No differences were observed in hypothalamic D(2) DA receptor mRNA abundance throughout the reproductive cycle, except for an increase in D(2) DA receptor mRNA within the INF of photorefractory hens. Also, a marked reduction in D(2) DA receptor mRNA was observed in the pituitary of incubating hens. Pituitary D(1D) DA receptor levels did not change when birds entered the incubating phase. Double ISH revealed that D(1D) and D(2) DA receptor mRNAs were co-expressed within neurons expressing VIP mRNA, predominantly within the lateral hypothalamus and INF. D(1D) DA receptor mRNA was more highly expressed than D(2) DA receptor mRNA. The present findings clearly demonstrate that the expression of stimulatory D(1) DA receptor mRNA in the hypothalamus increases in hyperprolactinemic incubating hens, whereas inhibitory D(2) DA receptor mRNA increases in the pituitary of hypoprolactinemic photorefractory hens. PMID- 12624533 TI - Mechanisms involved in glucagon-like peptide-1 (7-36) amide action on the rat hypothalamo-neurohypophysial system. AB - Glucagon-like peptide-1 (7-36) amide (tGLP-1) has been shown to modify the secretory function of the rat hypothalamo-neurohypophysial complex (HNC). However, mechanisms underlying this action are still unclear. Using explants containing the HNC obtained from euhydrated rats, possible interactions of tGLP-1 with angiotensin II (Ang II), forskolin-induced cAMP synthesis or calcium ions were investigated. In addition, explants taken from rats given 2% saline were used in order to examine whether chronic osmotic stimulation affects tGLP-1 action on vasopressin and oxytocin neurons. tGLP-1 did not modify Ang II- or forskolin-evoked hormone release. Incubation of the HNC in calcium-free medium inhibited the tGLP-1-dependent vasopressin/oxytocin secretion. Prolonged salt loading in vivo completely changed the neurohypophysial response to tGLP-1 in vitro; it did not only abolish the stimulatory effect of tGLP-1 on basal hormone release, but reduced K(+)-stimulated vasopressin/oxytocin secretion. Consequently, the neurohypophysial response to tGLP-1 may depend on the functional status of the HNC and on the presence of calcium ions, but not cAMP. PMID- 12624534 TI - Effects of galanin-like peptide on food intake and the hypothalamo-pituitary thyroid axis. AB - Galanin-like peptide (GALP) is a novel hypothalamic peptide synthesised in neurons in the arcuate nucleus which project to the paraventricular nucleus (PVN). GALP has recently been identified as an orexigenic peptide. In this study we aimed to further characterise the hypothalamic action of this peptide in energy homeostasis. Firstly, we investigated the orexigenic effect of GALP in the PVN and compared its effects with galanin and galanin 2-29. Secondly, we examined the effect of PVN administration of GALP and galanin on circulating thyroid stimulating hormone (TSH). PVN administration of GALP significantly increased the food intake of satiated rats 1 h after administration at doses of 0.3, 1 and 3 nmol. In comparison with paraventricular administration of galanin, GALP was a more potent orexigen, whereas galanin 2-29, the relatively selective GAL R2 agonist, had no effect on food intake. Both GALP and galanin administration (1 nmol) into the PVN significantly decreased the level of circulating TSH. To investigate the mechanism of these effects, we examined the effect of GALP and galanin application on neuropeptide release from hypothalamic explants in vitro. GALP peptide (100 nM) stimulated the release of the orexigenic peptide neuropeptide Y from hypothalamic explants and decreased the release of the anorectic peptide cocaine-and-amphetamine-regulated transcript, whereas galanin (100 nM) peptide had no significant effect on the release of either peptide. Both GALP (100 nM) and galanin (100 nM) inhibited the release thyrotrophin-releasing hormone. These data suggest that in the PVN, GALP may play a role in energy homeostasis by stimulating food intake and suppressing TSH release. PMID- 12624535 TI - Selective decreases in amphetamine self-administration and regulation of dopamine transporter function in diabetic rats. AB - The dopamine transporter (DAT) regulates extracellular dopamine DA levels and is an important site of action for amphetamine and cocaine. Amphetamine and cocaine increase extracellular levels of DA by acting on the DAT; thus, variations in DAT binding sites or activity might influence the action of some drugs of abuse. It was hypothesized that streptozotocin-induced diabetes decreases amphetamine self administration and that this behavioral change is accompanied by changes in DAT function. Separate groups of male rats responded to receive either amphetamine (0.03 mg/kg/infusion), cocaine (0.25 mg/kg/infusion), or food before and for 7 days after receiving streptozotocin. Rats were sacrificed and [(3)H]DA uptake and [(3)H]WIN 35,428 binding were measured in the striatum. In a second study, rats could self-administer one of several different doses of amphetamine (0.01-0.178 mg/kg/infusion) before and after receiving streptozotocin. In streptozotocin treated rats, a marked decrease in staining for insulin in pancreatic sections was paralleled by a more than doubling in blood glucose levels. Streptozotocin significantly decreased the number of amphetamine infusions without changing the number of cocaine infusions or food pellets received. Streptozotocin increased DA uptake (V(max)) 1.6- or 2.4-fold in rats that responded for food or amphetamine and increased 3-fold the K(m) for DA only in rats that responded for food; however, [(3)H]WIN 35,428 binding was not changed in any rat. In the second study, streptozotocin only decreased amphetamine self-administration thereby supporting the view that streptozotocin does not simply decrease the potency of amphetamine. These results demonstrate a selective decrease in amphetamine self administration in diabetic rats that was associated with increased DAT function in the striatum. Collectively, these studies suggest that insulin pathways in the brain may play an important role in regulating DAT activity and amphetamine action. PMID- 12624536 TI - Pharmacology and physiology of ovine corticosteroid receptors. AB - The aim of these studies was to characterize the ovine corticosteroid receptors (MR, mineralocorticoid receptors and GR, glucocorticoid receptors) in ovine hippocampus and brainstem. Adrenal-intact and adrenalectomized ewes were studied; adrenalectomized ewes were killed 47 +/- 9 h after steroid withdrawal, when symptoms of hypotension and/or hyperkalemia became evident. RT-PCR, immunoblotting and pharmacologic studies indicated the presence of both MR and GR in hippocampus and brainstem. Competitive binding studies using 3H-cortisol in brain tissue showed that the ovine MR binds cortisol, aldosterone and progesterone with equal affinity. Differences in receptor availability in intact and adrenalectomized ewes, along with determination of the binding affinity (K(d)) of MR and GR, suggested that MR occupancy is about 90%, whereas GR occupancy is about 30%, in normal animals. There was a significant increase in protein level of MR in brainstem, and the appearance of a higher molecular weight band for MR in hippocampus following steroid withdrawal, however no significant change in mRNA was detected by semiquantitative RT-PCR for either MR or GR in hippocampus or brainstem following steroid withdrawal. These studies suggest that physiological ligands of MR in the sheep brain include progesterone and cortisol, and that, as in other species, affinity of MR for cortisol is greater than that of GR. PMID- 12624537 TI - Temperature-induced changes in thyrotropin-releasing hormone sensitivity in carp melanotropes. AB - This study investigates whether thyrotropin-releasing hormone (TRH), alpha melanocyte-stimulating hormone (alpha-MSH) and N-acetyl beta-endorphin (NAc beta END), or the thyroid hormones thyroxine (T4) and 3,5,3'-triiodothyronine (T3) are involved in the physiological response to temperature changes in the poikilotherm common carp (CYPRINUS CARPIO). Carps were either subjected to a rapid cold exposure or acclimated over time to three different temperatures. Acute cold exposure did not influence blood plasma alpha-MSH concentrations. Acclimation to 15, 22 or 29 degrees C led to a temperature-dependent increase of both alpha-MSH and NAc beta-END plasma concentrations. Moreover, the in vitro sensitivity to TRH of melanotrope cells (that synthesise these peptides) also correlated positively with ambient temperature. Increased TRH activation stimulated processing of the precursor of alpha-MSH and NAc beta-END, resulting in increased release of both peptides and storage of a surplus of NAc beta-END within melanotropes. Plasma T4 levels were highest in carps acclimated to the intermediate temperature tested, and correlated strongly with hypothalamic TRH content. Plasma T3 levels were unaffected by ambient water temperature. We conclude that ambient water temperature influences the sensitivity of melanotrope cells to TRH in carps. This effect, however, is not due to acute temperature change, but evolves during the acclimation process of carps to a new temperature. PMID- 12624538 TI - The exacerbation of hippocampal excitotoxicity by glucocorticoids is not mediated by apoptosis. AB - Both endogenous and exogenous glucocorticoids (GCs) are known to cause apoptosis in a number of peripheral tissues and in some cases in the CNS. Additionally, GCs can exacerbate the neuron loss associated with such acute neurological insults as hypoxia-ischemia, excitotoxicity, and metabolic disruption. This exacerbation is accompanied by increased accumulation of glutamate in the synapse, excessive cytosolic calcium, and increased oxygen radical activity, markers usually attributed to pathways of necrotic cell death. It is also known that acute insults can involve apoptotic mediators. In this context, one outstanding question that has received little attention is whether the exacerbation of insult mediated cell death in neurons is apoptotic in mechanism. In this study we investigate whether the GC-mediated exacerbation of hippocampal excitotoxicity in culture involves apoptosis. Specifically, we show that while the magnitude of hippocampal neuron death caused by the excitotoxin kainic acid is indeed worsened in the presence of GCs, there is no evidence of increased markers of apoptosis. Specifically, we show that neither kainic acid nor GCs alone, or in combination, cause activation of caspase 3, a critical executor of insult-induced apoptosis. Furthermore, while kainic acid causes a significant incidence of apoptotic nuclear condensation, the incidence of this morphological indicator of apoptosis is not worsened by GCs. Thus, GCs appear to augment excitotoxic death in hippocampal neurons without augmenting the occurrence of apoptosis. We suggest that this finding is to be expected, given some energetic features of GC action and the energetic demands of apoptosis. PMID- 12624539 TI - In vivo activation of the interleukin-6 receptor/gp130 signaling pathway in pituitary corticotropes of lipopolysaccharide-treated rats. AB - Adrenocorticotropic hormone (ACTH) release from anterior pituitary corticotropes is greatly increased during peripheral inflammation induced by lipopolysaccharide (LPS) administration. Interleukin-6 (IL-6) is thought to participate in LPS induced ACTH release, but whether or not corticotropes are directly targeted by this cytokine is unclear. Therefore, we investigated the expression and activation of IL-6 signaling components in the pituitary of rats 2 and 4 h after administration of LPS (250 microg/kg). Intraperitoneal LPS treatment provoked the nuclear translocation of signal transducer and activator of transcription 3 (STAT 3) and Fos expression in the anterior pituitary lobe, as demonstrated by immunohistochemistry. By using in situ hybridization, we demonstrated that suppressor of cytokine signaling 3 (SOCS-3) and c-fos mRNAs were significantly induced by the LPS treatment in the anterior lobe of the pituitary. Dual in situ hybridization revealed that most corticotropes expressed IL-6 receptor and gp130 mRNAs, and that 2 h after LPS treatment, SOCS-3 and c-fos mRNAs were induced in corticotropes. Our results suggest that LPS-induced IL-6 could regulate the hypothalamo-pituitary-adrenal axis by directly targeting corticotropes during peripheral inflammation. PMID- 12624540 TI - Tamoxifen, a selective estrogen receptor modulator, reduces ischemic damage caused by middle cerebral artery occlusion in the ovariectomized female rat. AB - Previous work has demonstrated that physiological concentrations of 17beta estradiol can protect the female rat brain against middle cerebral artery occlusion (MCAO)-induced ischemic damage. The present study examined whether therapeutic doses of the clinically relevant selective estrogen receptor modulator (SERM), tamoxifen, can similarly protect the female rat brain against ischemic stroke damage. Adult female rats were bilaterally ovariectomized and implanted subcutaneously with either a placebo or tamoxifen time-release pellet (0.1, 0.8 or 2.4 mg/kg/day). One week later, the animals underwent permanent MCAO to assess the protective ability of the different tamoxifen doses on brain infarct size. As expected, MCAO produced a large infarct ( approximately 53%) of the affected cerebral hemisphere in placebo (control) animals. The 0.1 mg/kg/day dose of tamoxifen did not exhibit any significant protective effects, however; the 0.8 and 2.4 mg/kg/day doses of tamoxifen, which are in the therapeutic range, dramatically reduced infarct of the affected cerebral hemisphere ( approximately 70% reduction) as compared to the controls. The reduction of infarct size was primarily due to protection of two major structures, the cerebral cortex and striatum. Laser Doppler analysis further revealed that tamoxifen had no significant effect on cerebral blood flow either before or after MCAO, suggesting that tamoxifen protection is independent of cerebral blood flow changes. Further studies showed that tamoxifen pellets implanted at the time of MCAO did not reduce infarct size, suggesting that pretreatment with tamoxifen is necessary to observe a protective effect. These studies suggest that clinically important SERMs may have an additional unrecognized beneficial effect of protection of the female brain. PMID- 12624541 TI - Colocalization of progesterone receptors and thyroid hormone receptors alpha in the ovine diencephalon: no effect of estradiol. AB - Progesterone and thyroid hormones are key hormones in the control of reproduction in the ewe since both have been shown to inhibit GnRH secretion. GnRH neurons do not express progesterone receptors (PR) but half of them contain thyroid hormone receptors alpha (THRalpha), two nuclear receptors potentially able to act on gene transcription. PR and THRalpha distributions overlap in most regions of the ovine preoptic area (POA) and hypothalamus. To determine whether progesterone and thyroid hormones may also have common neuronal targets for the control of GnRH cell activity, we searched for coexpression of PR with THRalpha within neurons of the POA and hypothalamus of ovariectomized ewes treated with estradiol plus progesterone or with progesterone alone. Double-labeled cells were found throughout the medial POA, the periventricular part of the paraventricular nucleus (PVNpe), the arcuate nucleus (ARC) and the ventrolateral ventromedial nucleus (VMNvl). Colocalization ratios were not statistically different between steroid treatment groups. 84% of PR-immunoreactive cells in VMNvl, 90% in POA and ARC and 95% in PVNpe contain THRalpha. More than two-thirds of THRalpha immunoreactive cells contain PR in each area examined. This study provides evidence that progesterone and thyroid hormones may act within the same cells to modulate physiological functions such as reproduction. PMID- 12624542 TI - Circadian rhythms of neuroendocrine dopaminergic neuronal activity in ovariectomized rats. AB - Prolactin (PRL) secretion is inhibited by dopamine (DA) released from hypothalamic neuroendocrine neurons designated tuberoinfundibular (TIDA), tuberohypophyseal (THDA) and periventricular hypophyseal (PHDA) dopaminergic (DAergic) neurons. Since PRL is secreted in many physiological states with a circadian rhythm, the purpose of these experiments was to determine if patterns of neuroendocrine DAergic neuronal activity in rats are also circadian. The activity of neuroendocrine DAergic neurons, defined as DA turnover rate in nerve terminals and quantitated as the ratio of DOPAC (a primary DA metabolite) to DA content, was measured by high-performance liquid chromatography with electrochemical detection (HPLC-EC) in these populations of DA neurons of OVX rats. TIDA neurons exhibit a rhythm of activity in a light:dark cycle which free runs in constant dark (DD) and is entrained by light, indicating that TIDA neuronal activity is circadian. THDA and PHDA neurons also display daily rhythms entrained to a photoperiod and PHDA neuronal activity free-runs in DD with a period of approximately 24 h. However, a significant rhythm of THDA neuronal activity was not detected under DD. In the OVX rat, the activities of TIDA and PHDA neurons, but not THDA neurons, describe all the characteristics of a circadian rhythm as they are both entrained by light, but only TIDA and PHDA neurons maintain a significant rhythm of activity under DD. PMID- 12624543 TI - Lordosis of rats is modified by neurosteroidogenic effects of membrane benzodiazepine receptors in the ventral tegmental area. AB - Progestins modulate lordosis through actions in the ventral tegmental area (VTA). Whether neurosteroidogenesis of 5alpha-pregnan-3alpha-ol-20-one (3alpha,5alpha THP), involving mitochondrial benzodiazepine receptors (MBR), is important for lordosis was investigated. Ovariectomized (Ovx), hormone-primed rats (experiments 1, 3, 5, 6) and rats in behavioral estrus (experiments 2 and 4) were unilaterally infused via chronic guide cannula to the VTA with a MBR agonist, N,N-dihexyl-2-(4 fluorophenyl) indole-30-acetamide (FGIN 1-27) or antagonist 1-(2-chlorophenyl)-N methyl-N-(1-methylpropyl)-3-isoquinolinecarboximide (PK-11195). Experiment 1: Estradiol benzoate (EB)-primed (25 microg) rats administered 0 or 25 microg progesterone (P4) SC showed increased lordosis when infused with 5.0 microg FGIN 1-27 to the VTA; those administered 100 or 200 microg P4 SC exhibited greater lordosis when infused with 2.5 or 5.0 microg FGIN, relative to saline-infused rats. Experiment 2: Rats, near the termination of behavioral estrus, infused with 2.5 or 5.0 microg of FGIN 1-27 to the VTA, showed increased lordosis compared to that seen following vehicle administration. Experiment 3: EB-primed rats administered 200 or 500 microg P4 SC showed decreased lordosis when infused with 100, 200, or 400 ng PK-11195, relative to saline-infused rats. Experiment 4: Rats infused at the peak of behavioral estrus with 100, 200, or 400 ng PK-11195 to the VTA exhibited reduced lordosis compared to that seen following vehicle administration. Experiment 5: 3alpha,5alpha-THP (100 ng) infusions to the VTA reinstated lordosis of hormone-primed rats infused with PK-11195 (100 ng) to the VTA. Experiment 6: FGIN 1-27 (5.0 microg) and PK-11195 (100 ng) infusions aimed at the VTA respectively increased and decreased midbrain levels of 3alpha,5alpha THP compared to vehicle. Notably, the specific effects observed with infusions to the VTA were not seen with infusions to the control site, the substantia nigra. These data suggest that neurosteroidogenesis involving MBRs in the VTA mediates lordosis of hormone-primed or behavioral estrous rats. PMID- 12624544 TI - Effect of superovulation on the expression of tissue inhibitor of metalloproteinase-3 in the murine endometrium. AB - Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are important in the endometrial remodeling process during embryonic trophoblast invasion. Previous studies, using in situ hybridization, revealed that the predominant proteinase expressed by the implanting mouse embryo is matrix metalloproteinase-9 (MMP-9), and the major tissue inhibitor of metalloproteinases (TIMP) produced in the murine endometrium during this period is TIMP-3. Quantitative, competitive-reverse transcription-PCR was performed to evaluate the expression of TIMP-3 mRNA during the implantation period in the hyperstimulated murine endometrium from pregnant uteri of gestation day (g.d.) 6.0 and 7.0 after injection of 5 and 10 IU pregnant mare serum gonadotropin (PMSG). In PMSG groups, the expression of TIMP-3 mRNA, normalized by MMP-9 mRNA, showed a significant increase compared to the control group (natural coitus without PMSG injection) on g.d. 6.0 (p = 0.039) and an increasing tendency on g.d. 7.0. This study suggests that ovarian hyperstimulation by gonadotropin may have a detrimental effect on embryonic implantation possibly via an altered endometrial remodeling process by the increase in expression of TIMP-3 in the murine endometrium. PMID- 12624545 TI - Impact of epidermal growth factor and transforming growth factor beta-1 on the release of fibrinolytic factors from cultured endometrial and ovarian endometriotic stromal cells. AB - We have investigated whether there are any differences in the release of urokinase plasminogen activator (uPA) and its inhibitor (PAI-1) from cultured endometrial and endometriotic stromal cells, and whether the release is regulated by epidermal growth factor (EGF) or transforming growth factor beta1 (TGFbeta1). The cells were isolated from endometriomas and endometrium from women with and without endometriosis. After treatment with EGF or TGF and in untreated controls, incubated media collected at 0, 24, 48 and 72 h were analyzed by ELISA. Stromal cells from all three types of tissues released uPA and PAI-1, but the soluble receptor of uPA was not measurable in any group. The basal release of uPA and PAI 1 from endometriotic cells was higher than from endometrial cells. The uPA release in endometriotic cells was reduced with and without the addition of EGF (p < 0.05) or TGFbeta1 (p < 0.05). EGF increased the release of PAI-1 from stromal cells from women without endometriosis (p < 0.05) but decreased the release of PAI-1 from stromal cells from endometriotic women (p < 0.05). TGFbeta1 increased the release of PAI-1 from endometriotic cells (p < 0.05) but had no effect in endometrial cells. PMID- 12624546 TI - Clinical significance of high nm23-H1 expression in intraepithelial neoplasia and early-stage squamous cell carcinoma of the uterine cervix. AB - Seventy-two cervical pathology specimens, consisting of samples of cancer and high- and low-grade intraepithelial neoplasia, were used in a study exploring the involvement of nm23-H1/NDP kinase in carcinogenesis or recurrence. Additionally, the relationships between immunohistochemical expression of nm23-H1 and various clinicopathological variables for early-stage cervical cancer were evaluated. The nm23-H1 expression for high-grade cervical intraepithelial neoplasia and squamous cell carcinoma samples was significantly elevated when compared to low-grade analogs. Only deep stromal invasion was significantly associated with high nm23 expression. Moreover, a high cumulative recurrence hazard was demonstrated for the high nm23 expression group. In conclusion, high nm23-H1 expression may induce cellular proliferation for the progression from low- to high- grade intraepithelial neoplasia, with the subsequent emergence of invasive carcinoma as well as deep stromal invasion, and can be used as an indicator of prognosis. PMID- 12624547 TI - Thiazolidinedione inhibition of peritoneal inflammation. AB - Chemoattraction of macrophages into the peritoneal cavity is one of the important characteristics in patients with endometriosis. An inflammatory response is postulated to be responsible for infertility and pelvic pain associated with this syndrome. The present in vivo studies were designed to test if thiazolidinediones (TZDs), activators of peroxisome proliferator activated receptor gamma, could inhibit monocyte chemotaxis in a murine model. TZDs were first used as orally bioavailable insulin-sensitizing agents. They are currently under investigation in the treatment of inflammatory diseases, including arthritis or colitis. Intraperitoneal injection of thioglycollate was used to elicit high numbers of activated peritoneal macrophages in female mice. Concomitant peritoneal injection of ciglitazone, a member of the TZD family, significantly reduced the number of macrophages. When cultured and stimulated by tumor necrosis factor alpha, these peritoneal macrophages also secreted less RANTES and less IL-1beta protein. This animal model suggests that treatment of endometriosis patients with TZDs may diminish symptoms associated with intraperitoneal inflammation. PMID- 12624548 TI - Long-term administration of intravaginal dehydroepiandrosterone on regression of low-grade cervical dysplasia--a pilot study. AB - Although many dysplastic cervical lesions regress spontaneously, treatment is common due to concern for progression. Lesions persist or progress in women whose immune systems are unable to clear infection by human papillomavirus (HPV). Dehydroepiandrosterone (DHEA) is an adrenal steroid that has both immune modulatory and tumor inhibitory activity. A pilot study was conducted to examine the feasibility, safety and potential efficacy of intravaginal DHEA in women with low-grade cervical dysplasia. Twelve women with low-grade dysplasia, confirmed by colposcopic exam, were given 150 mg of intravaginal micronized DHEA daily for up to 6 months. Follow-up evaluations of the cervix were done at 3 and 6 months of use. DHEA, DHEA-S, androstenedione and testosterone levels were also measured. By the end of the study period, 10 of the 12 women (83%) had no evidence of dysplasia; the remaining 2 had normal colposcopic exams but cytology showing atypical cells of undetermined significance. There were no serious side effects. Androstenedione levels were elevated at 3 months, whereas testosterone levels were unchanged over the course of treatment. The results suggest that intravaginal DHEA is safe and well tolerated and may promote regression of low grade cervical lesions. Further study is needed to establish efficacy. PMID- 12624549 TI - Investigation of placental stem villi arteries in fetally growth-restricted pregnancies: a multivariate analysis. AB - Thirteen placentas were studied from 5 normal pregnancies and 8 from pregnancies complicated by fetal growth restriction (4 with present, 3 with absent, and 1 with reversed end-diastolic velocities at Doppler interrogation of the umbilical arteries). On immunohistochemically stained slides, the diameter (d) and the wall thickness (t) of the arterial vasculature of the stem villi were measured for a total of approximately 10,000 vessels. A multivariate 'mixed effect model' statistical analysis was performed using d and t as dependent variables and gestational age, delivery mode, fetal and placental weight, the degree of vascular collapse and Doppler blood flow patterns as independent variables. Gestational age, Doppler pattern and the degree of vascular collapse significantly affected both d and t, the mode of delivery influenced d while fetal and placental weights scarcely affected the dependent variables. The above parameters should therefore be taken into account when investigating placental stem vessel morphometry. PMID- 12624550 TI - Conventional and computerized antepartum telecardiotocography. Experienced and inexperienced observers versus computerized analysis. AB - The study assessed the agreement in cardiotocogram interpretation between the 2CTG computerized system and experienced and inexperienced observers involved in a telecardiotocography project called 'TOCOMAT'. Both observers and computer analyzed FHR baseline, FHR variability, and number of large accelerations and of decelerations. The k coefficient was calculated for the statistical analysis. The interobserver agreement about the evaluation of the FHR baseline and of the large accelerations was good; it was poor about the assessment of the FHR variability and the decelerations. Similar results were obtained for the agreement among observers and computer. The use of a computerized system provides exact values for most CTG parameters; the experts, however, should rely both on clinical features and on the computerized interpretation, to make decisions about management. PMID- 12624552 TI - Voiding symptoms in pregnancy: an assessment with international prostate symptom score. AB - OBJECTIVE: The aim of the study was to evaluate the voiding symptoms of pregnant women and to compare the symptoms amongst trimesters and controls by means of the International Prostate Symptom Score (IPSS). MATERIALS AND METHODS: Voiding symptoms of 256 pregnant and 230 non-pregnant healthy women hospitalized for other reasons were evaluated. Subjects who had a history of urological and neurological problems or previous pelvic surgery associated with urinary symptoms and those who had urinary tract infection were excluded from the study. The pregnant group was then subdivided into subgroups as to the trimesters of pregnancy. The IPSS assesses seven symptoms on a scale of 0 (no symptoms) to 5 (symptom always present) and was administered as a self-administered questionnaire with the assistance of a trained nurse. Symptom scores of each group and subgroup and the distribution of scores were assessed. RESULTS: The mean scores for all questions in the pregnant group were significantly higher than controls. The comparison of mean scores according to trimesters showed a significant difference for the questions of frequency, intermittency, urgency, weak stream, nocturia, quality of life and for the total scores. The distribution of the scores for each question showed that frequency and nocturia were the most prominent symptoms in pregnancy. The percentage of the respondents with an IPSS score of >7 was 47.6% and the percentage of a quality of life score of >/=4 (mostly dissatisfied) was 48.4%. CONCLUSIONS: Voiding symptoms during pregnancy are highly prevalent. These symptoms worsen as the pregnancy progresses. PMID- 12624551 TI - Comparison of neonatal effects of epidural and general anesthesia for cesarean section. AB - We assessed the influence of anesthetic technique for cesarean section on neonatal outcome. Thirty parturient women (ASA I/II) were randomly allocated into two groups. In Group GA general anesthesia was induced with 4 mg.kg(-1) thiopental and 1.5 mg.kg(-1) succinylcholine. In group EA epidural anesthesia was performed with 20 ml 0.375% bupivacaine through L(3-4) inter-space. 1-min Apgar scores were significantly higher in group EA (p < 0.001). Neurologic and Adaptive Capacity scores at 2 and 24 h were higher in group EA (p < 0.001). In terms of blood gas values, umbilical arterial pH and pO(2) values were higher in group EA (p < 0.05 and p < 0.001, respectively). The first breast-feeding intervals were found to be shorter in group EA (p < 0.001). We conclude that in terms of better Apgar and NAC scores, acid-base status and earlier initiation of breast-feeding, the epidural anesthesia may be preferred to general anesthesia in cesarean section. PMID- 12624553 TI - Ultrasound determination of chorionicity and perinatal outcome in twin pregnancies using dividing membrane thickness. AB - OBJECTIVE: To assess the value of ultrasound measurement of twin dividing membrane thickness in predicting chorionicity and perinatal outcome. To compare magnified with unmagnified images, and measurements taken with dividing membranes parallel and perpendicular to the ultrasound beam. METHODS: This was a prospective, observational cohort study of twin-pregnancy ultrasound examinations during the second and third trimesters with chorionicity confirmed by a pathologist. Ultrasound measurements of the dividing membrane thickness were recorded in 44 twin pregnancies using unmagnified and magnified images. Measurements were taken at various membrane sites either horizontal or vertical to the ultrasound beam, depending on the orientation of the dividing membrane. We compared the mean membrane thickness between monochorionic and dichorionic pregnancies as well as between pregnancies with good and adverse perinatal outcome. RESULTS: Membranes were significantly thicker in dichorionic pregnancies when compared with the monochorionic pregnancies (2.35 vs. 1.69 mm, p = 0.021). A cut-off point of >/=2.0 mm provided a sensitivity of 75.7%, a specificity of 85.7%, a positive predictive value of 96.6%, and a negative predictive value of 40.0%, for determining dichorionicity. Two of 7 monochorionic and 24 of 37 dichorionic pregnancies (59.1%) had good perinatal outcome. These pregnancies had significantly thicker membranes (2.44 vs. 1.96 mm, p = 0.022). Even among the dichorionic pregnancies, a membrane thicker than 2.2 mm was associated with a significantly higher proportion with good perinatal outcome (80.0 vs. 47.1%, relative risk 1.7, p = 0.036). Magnified images obtained with the ultrasound beam parallel to the membrane identified adverse outcome more accurately than unmagnified and perpendicular images. A cut-off point of >2.2 mm for membrane thickness was found to be the best predictor of good outcome. CONCLUSIONS: Membrane thickness can be used to identify chorionicity and adverse perinatal outcome, particularly if the ultrasound images are magnified and parallel to the ultrasound beam. PMID- 12624554 TI - Dysgerminoma with a slightly elevated alpha-fetoprotein level diagnosed as a mixed germ cell tumor after recurrence. AB - A pure dysgerminoma shows a normal serum alpha-fetoprotein level, and mixed germ cell tumors containing endodermal sinus tumor elements have elevated serum alpha fetoprotein levels, ranging from >100 to far higher than 1,000 ng/ml. A 40-year old woman was diagnosed as having a stage Ia pure dysgerminoma with a slight alpha-fetoprotein elevation (11 ng/ml), after a staging laparotomy, because we could not find any yolk sac element in the original tumor. After 44 months, she had a pelvic recurrent tumor with a significant elevation of the serum alpha fetoprotein concentration (1,520 ng/ml); histological examination of a needle biopsy specimen revealed a typical yolk sac tumor. Eventually, her initial tumor was diagnosed as a mixed germ cell tumor. The patient was successfully treated with seven courses of chemotherapy and has been disease free for 22 months. It is necessary to be aware of the possibility of a mixed germ cell tumor containing a yolk sac element, even when the alpha-fetoprotein level is only slightly elevated. PMID- 12624555 TI - Type 2 diabetes and preventive cardiology: talking the talk and walking the walk... PMID- 12624557 TI - A prospective study in primary care in patients without vascular disease comparing levels of coronary risk factors in those recommended for lipid-lowering drugs based on either absolute risk or absolute risk reduction. AB - In the United Kingdom, the current recommendation is that lipid-lowering drugs should be prescribed for primary prevention only to subjects with an absolute coronary risk (AR) greater than 15% in 5 years (i.e., myocardial infarction or angina). However, to achieve greater benefit it may be preferable to direct treatment to those patients showing the greatest absolute risk reduction (ARR). The aim of this study was to compare the characteristics of subjects eligible for lipid-lowering drugs based on the AR criteria or on an ARR of >4.45%. A prospective study was carried out over 29 months in primary care in a part of the United Kingdom with a prevalence of coronary disease nearly 20% above the national average. Risk factors were recorded in men and women aged 30-75 years who were being considered by their primary care physician for lipid-lowering drug therapy. Of the 2351 patients included in the study, 2139 (91%) and 101 (4.3%) were, respectively, below and above the criteria for treatment by both AR and ARR criteria. In 111 (4.7%) subjects, treatment was recommended based on only one of the criteria-82 on AR and 29 on ARR. Comparing these two groups, those treated on AR only were older (mean age 68.1 years [SD, 4.1] vs. 49.1 years [SD, 4.6]; p<0.0001) and had a lower total cholesterol (260 vs. 288 mg/dL; p=0.015); higher high-density lipoprotein cholesterol (50 vs. 43 mg/dL; p=0.003), lower low density lipoprotein cholesterol (160 vs. 184 mg/dL; p=0.03), a lower total to high-density lipoprotein cholesterol ratio (5.4 vs. 7.1; p<0.0001), and lower triglycerides (258 vs. 435 mg/dL; p=0.007). The AR group also had a higher mean systolic blood pressure (170.9 vs. 158.9 mm Hg; p=0.013), presumably an attribute of their greater age. Although the AR and ARR groups did not show a difference in the proportion of males or diabetics, there was a significantly greater proportion of smokers in the latter group (72% vs. 35%; p=0.001). In conclusion, treatment recommendations based on AR alone would result in nontreatment of young subjects with significant hyperlipidemia and at high relative risk of coronary disease, whereas lipid-lowering drugs would be given preferentially to patients whose main coronary heart disease risk factors are age and hypertension but not hyperlipidemia. By contrast, treatment recommendations based on ARR ensure that lipid-lowering drugs are directed to patients who will derive the most benefit. Furthermore, delaying treatment in younger subjects at high relative risk but not high AR results in their accumulating significant coronary risk in the years before their AR exceeds an arbitrary threshold before lipid-lowering drugs are prescribed. PMID- 12624556 TI - The effects of exercise on resting blood pressure in children and adolescents: a meta-analysis of randomized controlled trials. AB - Resting blood pressure in children and adolescents can track into adulthood. The purpose of this study was to use the meta-analytic approach to examine the effects of exercise on resting systolic and diastolic blood pressure in children and adolescents. Twelve randomized, controlled trials representing 16 outcomes in 1,266 subjects met the inclusion criteria. Reductions in blood pressure were approximately 1% and 3% for resting systolic and diastolic blood pressures, respectively. However, random-effects modeling using 5000 bootstrap confidence intervals revealed that neither result was statistically significant (systolic, x bar (x);+/-SEM=-1+/-2; 95% bootstrap confidence intervals=-2 to 0 mm Hg; diastolic, x+/-SEM=-2+/-1; 95% bootstrap confidence intervals=-3 to 0 mm Hg). The results of this study suggest that short-term exercise does not appear to reduce resting systolic and diastolic blood pressure in children and adolescents. However, a need exists for additional studies, especially in hypertensive children and adolescents. PMID- 12624558 TI - It is never too late: change in physical activity fosters change in cardiovascular risk factors in middle-aged women. AB - The purpose of the study was to determine the effect of physical activity, particularly change in physical activity over time, on cardiovascular risk factors in women. The 520 women in this analysis are part of an ongoing epidemiologic investigation of the effects of menopause on risk for cardiovascular disease; the investigation spans almost 20 years. The findings show that on average, physically active women have healthier risk factor profiles over time, and that as women change their activity level, their risk factor profiles change as well. Thus, for middle-aged women going through the menopausal transition, it is never too late to reduce their cardiovascular risk by increasing their activity level. PMID- 12624559 TI - The intestinal absorption of biliary and dietary cholesterol as a drug target for lowering the plasma cholesterol level. AB - Elevated plasma low-density lipoprotein cholesterol levels constitute a major risk factor for coronary heart disease. The plasma low-density lipoprotein cholesterol concentration is dictated partly by the efficiency of intestinal cholesterol absorption. The efficacy of treatments designed to block cholesterol absorption is partially offset to the extent that the liver compensates for the interruption to the enterohepatic movement of cholesterol by increasing the rate at which it synthesizes cholesterol. Currently, the most widely-used treatment for hypercholesterolemia is based on a class of agents (statins) that partially inhibit cholesterol synthesis within the body. Recent clinical trials with a unique, potent, and selective cholesterol absorption inhibitor (ezetimibe) used in combination with lower doses of various statins showed an additive reduction in plasma low-density lipoprotein cholesterol levels which equaled the reduction achieved with maximal doses of statins given alone. Combination therapy using a statin and this novel cholesterol absorption inhibitor represents an efficacious new approach to the treatment of hypercholesterolemia in the general population. PMID- 12624560 TI - Current recommendations for prevention of cardiovascular disease in patients with diabetes mellitus. Part II of II. AB - Patients with diabetes mellitus are at a high risk of developing cardiovascular disease, and therefore stand to benefit greatly from a preventive strategy. Recommendations regarding assessment and management of traditional risk factors are basically similar for diabetic and nondiabetic patients with several important differences. Several nontraditional risk factors also play a substantial role in the development of cardiovascular disease in diabetic patients, and need to be addressed if full preventive care is to be provided. In this second in a two-part series, we present current recommendations for reducing the risk of cardiovascular disease in the diabetic patient. PMID- 12624561 TI - Consumption of fish and fish oils and decreased risk of stroke. AB - Consumption of fish and fish oils was first associated with decreased risk of cardiovascular disease almost 50 years ago. Since then, a number of epidemiologic studies have evaluated whether their consumption is specifically associated with stroke. Ecologic/cross-sectional and case-control studies have generally shown an inverse association between consumption of fish and fish oils and stroke risk. Results from five prospective studies have been less consistent, with one showing no association, one showing a possible inverse association, and three demonstrating a significant inverse association. In the latest and largest of these, the Nurses Health Study, the relative risk of total stroke was lower, although not significantly so, among women who regularly ate fish than among those who did not. A significant decrease in the risk of thrombotic stroke (relative risk, 0.49; 95% confidence interval, 0.26-0.93) was observed among women who ate fish at least two times per week compared with women who ate fish less than once per month, after adjustment for age, smoking, and other cardiovascular risk factors; a nonsignificant decrease was observed among women in the highest quintile of long-chain omega-3 polyunsaturated fatty acid intake. No association was observed between consumption of fish or fish oil and hemorrhagic stroke. These data support the hypothesis that consumption of fish several times per week reduces the risk of thrombotic stroke but does not increase the risk of hemorrhagic stroke. PMID- 12624562 TI - Obesity and the risk for cardiovascular disease. AB - Coronary heart disease (CHD) is the leading cause of mortality in the United States. Hypertension, diabetes mellitus, hypercholesterolemia, and smoking have all been directly related to CHD. Obesity is on the rise in the United States and has also been associated with CHD. This review clearly establishes obesity as an independent risk factor for CHD as demonstrated by the Framingham Heart Study, Nurses Health Study, Buffalo Health Study, and the Cancer Prevention Study II. Morbid obesity was found to correlate with a significant risk of mortality from CHD, especially in young men. Prevention of obesity, and therefore reduction in risk from cardiovascular disease, is paramount in the management of obesity. New approaches to behavioral, medical, and surgical management of obesity are reviewed, including thalidomide, an antiangiogenic agent. A primary and secondary prevention model details a multidisciplinary approach to reducing risk in obesity. PMID- 12624563 TI - Psychologic treatment for hypertension can be efficacious. AB - The traditional view of psychologic treatments and their efficacy for treating primary hypertension is that they may be of some benefit for borderline hypertension and that they can be potentially useful adjuncts to medical treatment. This review challenges such cautious conclusions by critically analyzing previous reviews and by also considering innovative, newer clinical trials. One reason for cautious conclusions has been selective reviewing and a lack of well-designed trials. Measurement and study protocol differences as well as differences in treatment modality have all been shown to have major effects on observed outcomes. Studies that promise better results have the following characteristics: 1) use of 24-hour ambulatory monitoring as an end point to rule out white coat hypertension that does not need treatment in the first place; 2) inclusion of patients with truly elevated blood pressure at pretreatment; and 3) multicomponent treatments that are tailored to individual patient needs. PMID- 12624564 TI - ALLHAT and AFFIRM. PMID- 12624565 TI - Hormone replacement therapy and primary prevention: alas, again... PMID- 12624566 TI - The Dietary Intervention Study in Children (DISC): progress and prospects for primary prevention. PMID- 12624567 TI - Predictors of quality of life in women 1 year after myocardial infarction. AB - In an analysis of 30 African American and Caucasian women, the authors describe changes in the various quality-of-life domains (health, family, socioeconomic, psychosocial, and spiritual) and predictors of quality of life for a group of women more than 1 year post-acute myocardial infarction (MI). Data reported represent three recovery points post-MI (before hospital discharge, 6-weeks post MI, and 12-months post-MI). Matched pair t tests indicated significant changes in all five domains of quality of life/cardiac, and through a multiple regression analysis 45% of the variance in quality of life could be explained by mood states and social support. The finding that social support and mood states were the only predictors of quality of life for this cohort of women at 1-year post-MI suggests that health care professionals should mobilize and reinforce social support networks and make mental health referrals for the most vulnerable women. Further study should be conducted on the unique needs of African American women, since insufficient numbers were available at 1 year to determine their unique patterns of recovery. PMID- 12624568 TI - Understanding the essentials of blood lipid metabolism. AB - The authors present an introduction to the basics of lipid metabolism including an overview of the structure and function of lipoproteins and a description of the pathways of lipid metabolism. Dyslipidemia is an important risk factor in the context of cardiovascular disease, and appropriate intervention can have a significant impact on clinical outcomes. The information presented herein will help to provide a foundation of knowledge on which to base the assessment and treatment of dyslipidemic patients. A better understanding of lipid metabolism will help health care professionals to provide better care in the realm of dyslipidemia management. PMID- 12624569 TI - Psychosocial issues in patients with congestive heart failure. AB - Psychosocial issues are important variables that need to be addressed in patients with congestive heart failure (CHF). Unfortunately, these issues are often overlooked. Depression and lack of social support have been shown to have a negative impact on patients with CHF. Patients with CHF who are depressed or who lack social support have been shown to have increased morbidity and hospital readmission rates, to be less adherent to their medical regimen, and to have an overall increase in cost of care. The variables are often interrelated, as high levels of social support may lessen the impact of depression on mortality. In addition, certain biologic factors may influence the impact of psychosocial factors in patients with CHF. This review addresses the effects of depression, treatment adherence, and social support in patients with CHF and suggests interventions targeted to these problems. Health care professionals must assess these issues in all patients with CHF, address their specific needs, and intervene appropriately when warranted. PMID- 12624571 TI - The role of infection in the pathogenesis of cardiovascular disease. AB - Atherosclerosis is a complex, multifactorial disease. Recently, research has intensified to identify the role of various infections in the pathogenesis of atherosclerosis. Specific agents have been proposed as direct initiators or accelerators of atherosclerosis, while other infectious agents have been proposed as accelerators of atherosclerosis through nonspecific stimulation of the inflammatory cascade. Recently, the total pathogen burden concept has suggested that while each specific infection contributes only slightly to the pathogenesis of atherosclerosis, the cumulative effects of infectious agents contribute greatly. Several randomized trials evaluating antibiotic therapy in the prevention of cardiovascular events have now been completed, although results have been conflicting. This manuscript summarizes current understanding of the role of infectious agents as a trigger of inflammation, as a contributor to atherosclerosis, and the potential role of antibiotic therapy in the treatment of atherosclerosis. PMID- 12624570 TI - A summary of results of the Dietary Intervention Study in Children (DISC): lessons learned. AB - Prevention of cardiovascular disease must begin in childhood, preferably before risk factors develop. Elevated low-density lipoprotein cholesterol levels in children are likely to track over time and become high-risk levels in adults. The Dietary Intervention Study in Children (DISC) was a multicenter, collaborative randomized trial in pre-adolescent children designed to test the efficacy and safety of a dietary intervention to lower saturated fat and cholesterol intake among growing children with elevated low-density lipoprotein cholesterol. Numerous DISC results, which include findings on lipids-lipoproteins, genetics, and nutrient adequacy, as well as descriptions of the behavioral intervention strategies, have been reported. A summary of practical findings and their potential clinical applications have not previously been published. Highlights of key lessons learned from DISC and translational applications of potential interest to nurses and other health care providers are presented. PMID- 12624572 TI - Cardiovascular nursing in Israel. AB - Cardiovascular (CV) nursing as an entity in Israel dates back to 1952, when the nurses in Tel-Hashomer hospital took care of postoperative heart surgery patients. The first intensive cardiac care units (ICCUs) were established in 1971. In 1982, the first ICCU course was established in Tel-Hashomer hospital nursing school. Today, most of the nursing staff in Israels ICCUs are graduates of ICCU courses. The nurses professional society, the Society for Nursing of Israel, was established in 1947. In 1989 the Society for Advancement of Cardiac Nursing in Israel (SACN) was established. The main goals of the society were: the exchange of CV nursing knowledge, CV nursing research, CV nursing education in nursing schools, education of nurses in other departments in the care of the cardiac patient, and CV nursing education in the community. The CV nurse takes a large role in the total care of the cardiac patient, which includes rehabilitation within the hospital and in the ambulatory setting and coordination of nursing in national and international multicenter clinical trials. In collaboration with the Ministry of Health Nursing Division, Israeli CV nurses participate in national and international projects to: develop and upgrade nursing education; train new CV nurses; develop, review, and revise nursing protocols and guidelines; and establish new, more advanced ICCUs in underdeveloped areas within Israel and around the world. Our vision for the future development of CV nursing in Israel includes coordination and management roles in the hospital setting, and the establishment and management of home-care programs. PMID- 12624573 TI - Aldosterone receptor antagonists: focus on eplerenone. AB - Despite the development of hypertension treatment guidelines, blood pressure control in the general population remains inadequate, indicating the need for ongoing re-evaluation of treatment strategies to further improve blood pressure control. Hypertension results from alterations in cardiac output and/or peripheral resistance. The renin-angiotensin-aldosterone system may be responsible, at least in part, for these alterations. Despite pharmacologic intervention with angiotensin-converting enzyme inhibitors and angiotensin type-1 receptor antagonists, aldosterone continues to be produced. Therapeutic modalities for treating hypertension directed toward antagonizing aldosterone might more effectively control blood pressure. Eplerenone, a new selective aldosterone receptor antagonist, recently received approval from the US Food and Drug Administration for the treatment of hypertension, either alone or in combination with other antihypertensive agents. The objective of this review is to summarize the renin-angiotensin-aldosterone system, emphasizing the role for aldosterone antagonism in the management of hypertension, with a focus on eplerenone. PMID- 12624574 TI - When does ST-segment depression in the presence of a narrow QRS tachycardia signify ischemia? PMID- 12624575 TI - Conversations that matter-the road to patient safety. PMID- 12624576 TI - A spectrum of clinical issues in geriatric cardiology. PMID- 12624577 TI - Clinical correlates of isolated left ventricular diastolic dysfunction among hospitalized older heart failure patients. AB - Heart failure due to isolated left ventricular diastolic dysfunction (LVDD) is common among older adults. The purpose of this study was to develop a model based on admission clinical features to predict isolated LVDD in older adults hospitalized with heart failure. The authors studied 394 Alabama Medicare beneficiaries discharged with a primary discharge diagnosis of heart failure. All patients had data on LV function determined by echocardiography performed during the index hospitalization. Isolated LVDD was defined by LV ejection fraction >/=45% or LV function described as normal. The authors used all subsets multiple logistic regression analyses to examine various models predicting isolated LVDD. Patients had a mean (+/-SD) age of 78 (+/-7) years and 58% were women. One hundred thirty nine (35%) patients had isolated LVDD. More women (47%) than men (21%) had isolated LVDD (p<0.001). Variables independently associated with isolated LVDD in the selected 12-predictor model were female sex (odds ratio [OR]=3.10; 95% confidence interval [CI]=1.86-5.15), pulse 100 beats/min or greater (OR=0.43; 95% CI=0.25-0.72), systolic blood pressure (OR=1.01; 95% CI=1.00-1.02), diastolic blood pressure (OR=0.98; 95% CI=0.96-0.99), pulmonary vascular congestion by chest x-ray (OR=0.51; 95% CI=0.30-0.88), coronary artery disease (OR=0.54; 95% CI=0.32-0.92), cardiomyopathy (OR=0.21; 95% CI=0.10-0.47), and admission to a teaching hospital (OR=0.36; 95% CI=0.15-0.88). The authors concluded that the proportion of patients with isolated LVDD among this cohort of elderly hospitalized heart failure patients was low and was associated with few clinical characteristics. Except for female sex and systolic blood pressure, all model variables were negatively associated with isolated LVDD. PMID- 12624578 TI - Enhanced external counterpulsation in the management of angina in the elderly. AB - This study was undertaken to determine whether enhanced external counterpulsation is a safe and effective treatment for angina in octogenarians. In this prospective observational study, demographic and clinical outcome data on patients consecutively enrolled in the International EECP Patient Registry was examined. Of the 3037 patients analyzed, 249 (8%) were >/=80 years old. Octogenarians were more likely to be female and have a history of congestive heart failure (41% vs. 29%; p<0.001). They were less likely to have had previous revascularization. Fewer patients in the octogenarian group (76% vs. 84%; p<0.01) completed a course of treatment. Of those octogenarian patients who completed treatment, 76% reported a reduction in angina and quality of life improved significantly. Adverse events related to treatment were low. At 6-month follow up, 81% reported maintenance of angina improvement. Thus, enhanced external counterpulsation is a low-risk intervention that offers octogenarians the ability to return to more normal activity and a better quality of life. PMID- 12624579 TI - Strength training and hemodynamic responses to exercise. AB - Sixty-two older adults (68.4+/-6 years) were randomized into a control group (n=16), low-intensity exercise group (n=22, LEX; 50% 1-repetition maximum; 13 repetitions per exercise), or a high-intensity exercise group (n=24, HEX; 80% 1 repetition maximum; eight repetitions per exercise) group. Subjects trained for 24 weeks, performing 1 set for each of 13 exercises. Subjects performed a graded exercise test (GXT) at baseline and after 24 weeks of strength training. Heart rate, systolic, and diastolic blood pressures were measured at rest, during the GXT and 1, 3, and 5-min post-GXT. Mean arterial pressure was also calculated. Post-training, time to peak systolic and diastolic blood pressures and heart rate was increased in the LEX and HEX groups by 22.9% and 44.5%, respectively (p<0.05). Diastolic blood pressure, heart rate, and mean arterial pressure during the GXT were significantly decreased (p<0.05) in the HEX group following training. Post-GXT heart rate was lower in the HEX and LEX groups compared to control (p<0.05) indicating a more rapid recovery. Resistance exercise prolongs the onset of peak cardiovascular responses, decreases the cardiovascular response to exertion, and improves recovery from maximal exertion. PMID- 12624580 TI - Dobutamine stress echocardiography in preoperative and long-term postoperative risk assessment of elderly patients. AB - The authors reviewed all negative preoperative dobutamine stress echocardiograms (DSEs) performed over a 3-year period to determine the value of negative DSE for preoperative risk assessment in elderly patients. All patients with negative DSE performed for preoperative evaluation were followed. Cardiac event rates during and after the operative procedure were determined for hard end points (nonfatal myocardial infarction, cardiac death) and soft end points (emergency room visits, hospitalization for unstable angina, congestive heart failure, coronary angioplasty, coronary artery bypass graft surgery). Results noted that DSEs were negative for ischemia in 82 preoperative evaluations. Group 1 (age >/=65; n=41) had hard and soft event rates per patient/year of 0.97% and 7.3%, while group 2 (age <65; n=41) had hard and soft event rates per patient/year of 0.81% and 10.8%. There were no significant differences in event rates between the two groups (p=NS). In conclusion, the authors found that negative DSEs predict low cardiac event rates in elderly patients during the perioperative and long-term postoperative periods, which are not significantly different from the cardiac event rates in a younger cohort. PMID- 12624581 TI - Proper planning for end-of-life medical care; confronting the near irrelevance of currently used living wills. PMID- 12624582 TI - Transesophageal and transpharyngeal echocardiographic detection of the extracranial segments of the left vertebral artery. PMID- 12624583 TI - Minimal requirements for interpreting Wenckebach (Mobitz I) A-V block. PMID- 12624585 TI - [Magnetic resonance cerebral blood volume maps. Comparison with histologic findings in different types of brain lesions]. AB - Recent developments in magnetic resonance (MR) have made it possible to obtain measurements of the microvasculature within brain lesions. Cerebral blood volume (CBV) maps calculated from dynamic contrast-enhanced MR imaging are particularly sensitive for depicting the microvasculature, and can enable the detection of neovascularization as well as its quantification in relative terms. The purpose of the present work is to compare the results of CBV maps calculated from MR imaging with those from histologic examination of the same region of interest: the biopsy site. Nineteen patients with brain lesions were studied (18 brain tumors and one case of multiple sclerosis). All patients underwent stereotactic biopsy, and calculation of CBV was performed from perfusion MR imaging. Three histopathologic parameters were assessed: the number of vessels (vessel density), the vessel size and the surface area filled by vessels (%). We observed a statistically significant correlation between the vessel density and the CBV, which is consistent with some previous publications. A noninvasive imaging method for characterizing the functional properties, especially hemodynamic activity, of malignant processes seems to be of great benefit to clinical practice. PMID- 12624586 TI - [MRI of cerebral cortical dysgenesis]. AB - PURPOSE: Cortical cerebral dysgenesis is a heterogeneous disorder of cortical development and organization. It is well known by neuropathologists and is becoming more important in the clinical setting, mainly due to MRI. Our purpose is to report the MRI features of cortical dysplasia in 30 patients and to propose a simple classification with review of the literature. PATIENTS AND METHODS: It is a retrospective study of the MRI features of 30 cases of cortical dysgenesis. All patients were evaluated by electroencephalography and brain MRI using T1 and T2 weighted, inversion recovery T1 weighted, FLAIR and MPR 3D sequences with multiplanar reformations. RESULTS: Twenty-nine patients were referred to the neurology department because of refractory epilepsy and one patient for mental retardation. Our patients were distributed in 4 groups: diffuse forms represented by agyria-pachygyria (4 cases), polymicrogyria (one case), and nodular heterotopias (5 cases), local bilateral forms represented by bilateral polymicrogyria (11 cases), unilateral forms represented by hemi-megalencephaly (one case), and focal cortical forms found in eight cases. CONCLUSION: Neuroblast migration disorders are better known, mainly due to high resolution MRI techniques that by the quality of multiplanar study and a better differentiation between gray and white matter, allow a more precise diagnosis. PMID- 12624587 TI - [Lyrical and musical auditive mental imagery in functional MRI]. AB - PURPOSE: to explore with functional MRI cerebral areas involved in musical and lyrical sounds signal processing with the mental imagery method. MATERIAL: and METHODS: nine volunteers (mean age: 27 years old) underwent functional MRI with BOLD contrast at 1.5 T. Box-car paradigms of partial recollections of musical or lyrical memories tasks were performed. Statistical correlations mappings were calculated and superimposed on previously realigned anatomical reference imaging to observe activated cerebral areas. RESULTS: all except one subjects had activation areas in primary and secondary auditive cortices in the temporal Heschl gyrus and the Planum Temporale, unilaterally (n=2) or bilaterally (n=6) during both mental tasks. Contralateral activation improvement was observed in 4 cases when the lyrical tasks were performed. Temporal and insular regions involved in language processing were observed in eight of nine subjects. CONCLUSION: auditive mental imagery can show in functional MRI cerebral areas involved in auditive functions and some of the areas involved in language processing. PMID- 12624588 TI - Diffusion weighted MR imaging on a low-field open magnet. Comparison with findings at 1.5T in 18 patients with cerebral ischemia. AB - Diffusion-weighted MR imaging (DWI) is particularly sensitive for the detection of acute stoke. Until recently, DWI was performed with EPI technology. We compared 18 patients with clinical suspicion of acute stroke on a standard 1.5T unit and an open low-field MR scanner. Eighteen patients with 20 lesions of acute stroke were studied retrospectively with DWI and ADC mapping on both systems. The technique used was a rotating fast-spin echo T2 at low-field and an EPI sequence at 1.5T. Both examinations were performed within 24 hours and analyzed by two neuroradiologists. We obtained the same results on DWI sequences on both systems, regarding high intensity lesions on DWI. Interpretation of the ADC maps proved to be difficult on low-field MR near the lateral ventricles (3/18). We experienced the same difficulty of interpretation at low and high field in the cerebellum, in the temporal fossa and in cortex situated near bone, due to susceptibility artifacts. Chronic lesions were better visualized at low than at high field. In our opinion, DWI on a low-field open MR scanner is a good technique to evaluate subacute stroke and was as reliable as when performed on a 1.5T MR system. PMID- 12624589 TI - [Electronic imaging with photo-realistic rendering for neuroanatomy teaching. Methods and preliminary results]. AB - Advances in software, networking, and imaging technology provide a unique opportunity for developing information systems in anatomy that can deliver relevant knowledge to the clinicians, researchers, educators, and students. Recent software tools initially produced for graphic imaging are now available in the medical graphic design field. The authors describe an original method they used to create electronic images of the central nervous system and its coverings with photo-realistic rendering. They present preliminary results and discuss the potential of this new technique as a teaching tool for neuroanatomy. PMID- 12624590 TI - [Value of single photon emission computed tomography in the characterization of fronto-temporal dementia subtypes]. AB - DFT (fronto-temporal dementia) was diagnosed in 33 patients (17 males and 16 females), mean age 74.9 years, using the criteria from Neary et al. (1998). The severity of the disease was evaluated based on the presence or absence of 4 clinical groups of signs (behavioral, affective, neurological signs and language) allowing the calculation of a global clinical ratio on 20. A CT-scan and a Single Photon Emission Computed Tomography (SPECT) scan using Tc99m-ECD with quantification of brain perfusion were then performed. This study confirmed the classical clinical forms of DFT: a psycho-affective form (21%), where negative symptoms are predominant, and a behavioral disinhibited form (9%). Most of our cases (40%) corresponded to a global form. Finally we also identified a fourth clinical form: cognitive form because of predominance of language disturbances (30%). Concerning SPECT, we found 4 topographic forms: frontal global (46%), frontal superior (24%), frontal inferior (15%) and frontal unilateral (12%). In the frontal inferior form, behavioral disturbances were correlated to the intensity of frontal inferior hypoperfusion, corresponding to an orbito-frontal dysfunction. In the frontal superior form, psycho-affective disturbances (apathy) were correlated with frontal superior hypoperfusion (anterior cingulum and superior dorso-lateral cortex). Finally, in global and frontal inferior SPECT forms, language disturbances were correlated with a frontal inferior hypoperfusion (prefrontal and inferior left frontal cortex). PMID- 12624591 TI - [Diagnosis of periventricular ependymal enhancement in MRI in adults]. AB - Periventricular enhancement in adults at MRI is a significant finding since it often indicates the presence of an underlying disease requiring prompt medical attention. From a review of patients with periventricular enhancement, the main imaging features based on the underlying infectious or tumoral etiology will be described. The presented differential diagnosis is based on the immune status of the patient, type of enhancement, and response to a trial therapy. In immunocompromised patients, the main considerations are lymphoma and viral ependymitis. The pattern of enhancement is important. The presence of thin linear enhancement suggests a viral etiology (cytomegalovirus or varicella-zoster virus) that can be confirmed at CSF evaluation whereas the presence of nodular enhancement suggests a diagnosis of primary CNS lymphoma that can be confirmed by the presence of lymphomatous cells in the CSF or, more frequently, at stereotactic surgical biopsy performed after failure of response to anti toxoplasmosis treatment. The presence of band enhancement is less specific and can be seen with viral, lymphomatous and even tuberculous involvement. In immunocompetent patients, a clinical context of infection will suggest bacterial or tuberculous ventriculitis and the presence of cystic lesions will suggest cysticercosis; in the absence of constitutional symptoms, the presence of nodular enhancement will suggest a tumoral process (lymphoma, ependymoma, germ cell tumor, or metastases). Rarely, linear enhancement will be due to sarcoidosis or Whipple's disease. PMID- 12624592 TI - [Intracerebral hematoma associated with reduced apparent diffusion coefficient mimicking acute stroke]. AB - We report the case of a 67 year old patient admitted at our institution for acute onset of left hemiplegia. MRI was done 2 h 30 after symptom onset. Diffusion weighted images showed a hyperintense lesion in the right basal ganglia region with restricted apparent diffusion coefficient (ADC=428. 10(-6) mm(2)/s), a 50% decrease in value compared to the normal left side, consistent with acute ischemia. The lesion was hyperintense and moderately heterogeneous at FLAIR imaging, best seen on echo planar T2W images, with hypointense rim consistent with magnetic susceptibility artifact. The appearance and location of the lesion suggested the possibility of hematoma, which was confirmed at CT. Interpretation of ADC values must be performed in correlation with results at imaging including sequences sensitive to magnetic susceptibility artifacts such as echo planar T2*W and T2W sequences in order to exclude the possibility of underlying hematoma. PMID- 12624593 TI - [Siderosis of the brain and spinal cord. Report of two cases]. AB - Two cases of superficial siderosis of the brain and spinal cord with cochleovestibular and cerebellar symptoms are diagnosed on brain and spinal MRI scans. Low signal intensity lines are noted on the surface of the brainstem, cerebellum, spinal cord and within the interhemispheric and sylvian fissures. In one case, no brain or vascular malformation is identified; in the second case, two cavernous angiomas are noted on the MRI study. 3D CISS may visualize thickening of the cochleovestibular nerve. PMID- 12624594 TI - Blood-borne viral diseases and the surgeon. PMID- 12624596 TI - Is a needle in the haystack worth the trouble? PMID- 12624597 TI - Infection control during gastrointestinal endoscopy. AB - Infection-control issues during gastrointestinal endoscopy, which are becoming increasingly important, can generally be divided into three major areas: (1) infectious complications resulting from a patient's own microbial flora (autologous), (2) infections transmitted from patient to patient by way of the endoscope (exogenous), and (3) infections transmitted between the patient and the health-care provider. The mean frequency of postprocedure bacteremia ranges from 0.5% for flexible sigmoidoscopy to 2.2% for colonoscopy, 4.2% for esophagogastroduodenoscopy, 8.9% for variceal ligation, 11% for endoscopic retrograde cholangiopancreatography, 15.4% for variceal sclerotherapy, and 22.8% for esophageal dilation. Although postprocedure bacteremia is not uncommon, it seldom results in infectious complications. Exogenous infections transmitted during endoscopy, which are extremely rare, generally result from failure to follow accepted guidelines for the cleaning and disinfection of gastrointestinal endoscopes, underscoring the importance of meticulous attention to endoscope reprocessing. Finally, although the risk of patient-staff transmission of infection is also rare, standard infection-control recommendations are important in protecting both patients and health-care providers. PMID- 12624598 TI - Induction of apoptosis and ubiquitin hydrolase gene expression by human serum factors in the early phase of acute myocardial infarction. AB - The physiologic events leading to apoptosis in myocardial infarction and the molecules involved in the death process have not been clarified unequivocally. We developed a method to search for serum factors that induce apoptosis of human cells, using serum obtained from patients within 1 day of the onset of acute myocardial infarction (AMI). Serum factors were found to have the ability to increase the caspase-3 activity levels in human RSa cells, which are susceptible to apoptosis inducers. The factors obtained from AMI patients by elution at about 0.5 mol/L KCl from a dye-ligand column were named AMI-SFs (serum factors from AMI). Electrophoretic analysis showed DNA fragmentation in AMI-SF-treated RSa cells, but not in RSa cells treated with fractions from AMI patients 1 week after clinical onset of illness. AMI-SF-induced DNA fragmentation was also demonstrated by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling analysis, whereas a suppression of fragmentation was seen in RSa cells treated with AMI-SFs in combination with a caspase-3 inhibitor. The increase in caspase-3 activity was not inhibited by neutralizing antibodies to tumor necrosis factor-alpha, interleukin-6, human interferon-beta, or interferon gamma. Polymerase chain reaction-based messenger RNA differential display and Northern blotting revealed an increase in the messenger RNA expression level of human ubiquitin hydrolase in AMI-SF-treated RSa cells. Antisense oligonucleotides for ubiquitin hydrolase inhibited the increase in caspase-3 activity. These findings suggested that serum from AMI patients in the acute phase contains factors that induce apoptosis, possibly by inducing the expression of the ubiquitin hydrolase gene, at least in the human cells tested. PMID- 12624599 TI - Quantification of the cell-cycle inhibitors p27(Kip1) and p21(Cip1) in human atherectomy specimens: primary stenosis versus restenosis. AB - Proliferation, a key determinator of vascular proliferative diseases, is dependent on cyclin/cyclin-dependent kinase (CDK) complexes, which are controlled by cyclin-dependent kinase inhibitors (CKIs) such as p27(Kip1) and p21(Cip1). Both have prognostic significance in various human malignancies. We have determined the levels of p27(Kip1) and p21(Cip1) in human directional coronary atherectomy specimens of primary lesions (n = 15) and lesions of in-stent restenosis (n = 18) in comparison to those of other vascular regions and have correlated CKI levels with clinical data. Quantitative immunoblotting demonstrated low expression of p27(Kip1) in primary lesions (5.9 +/- 0.5 ng/mg protein) compared with that in aorta (14.9 +/- 0.9 ng/mg), internal mammary artery (16.7 +/- 1.1 ng/mg), and carotid artery thrombendarterectomy specimens (16.5 +/- 1.7 ng/mg). Similarly, p27(Kip1) levels in lesions of in-stent restenosis were found to be significantly reduced (6.3 +/- 1.1 ng/mg; mean time of restenosis development 367 +/- 61 days). p27(Kip1) levels did, however, not have prognostic significance for the development of restenosis, and expression levels of proliferating cell nuclear antigen and CDK2 were similar in all groups examined, indicating low proliferative activity. Clinically, p27(Kip1) was not of value in predicting the development of restenosis. Furthermore, p27(Kip1) tissue levels were not increased in statin-treated patients, implying that the favorable effect of these drugs is not a result of p27(Kip1) stabilization. However, the relative content of p21(Cip1) was found to be significantly up-regulated in restenosis compared with that in primary lesions (225%) and the other vascular regions. Our data imply that negative-feedback mechanisms are still intact in coronary proliferative disease, thereby contrasting the finding of deregulated proliferation in malignancies. PMID- 12624600 TI - Glioma apoptosis induced by macrophages involves both death receptor-dependent and independent pathways. AB - Apoptosis of glioma may represent a promising intervention for tumor treatment. Macrophages are able to induce apoptosis in a number of tumor cells, including glioma. It is known that apoptosis of cells is executed on either a death receptor-dependent or independent pathway. Whether and how apoptosis of glioma cells induced by activated macrophages is involved in these two pathways simultaneously are not known. Using in vitro and in vivo experimental models, we investigated Bcl-2 system and Fas/FasL channel, representing the death receptor dependent and independent pathways, respectively, in glioma cells treated with the supernatant from the activated macrophages, which was rich in tumor necrosis factor-alpha and interferon-gamma. We found that levels of Fas and FasL were up regulated both in vitro and in vivo, accompanying an increase in the expression of caspase-8. The number of apoptotic cells was also increased significantly, although the percentage of death cells exceeded the number of tumor cells positive for Fas or FasL. It was also evident that the expression of Bax was increased, whereas the level of Bcl-2 was decreased, in glioma cells treated with the supernatant from the activated macrophages. The alteration of molecules related to both death pathways led to apoptosis of glioma and the inhibition of xenograft glioma growth in mice. Apoptosis of glioma induced by the activated macrophage is executed by way of both death receptor-dependent and independent pathways, and such an apoptosis-induced approach can effectively inhibit the growth of glioma in vivo. PMID- 12624601 TI - High blood glucose and osmolality, but not high urinary glucose and osmolality, affect neuronal nitric oxide synthase expression in diabetic rat kidney. AB - We recently demonstrated that neuronal nitric oxide synthase (nNOS) messenger RNA (mRNA) is markedly increased in the kidneys of diabetic rats and water-deprived rats. It can be inferred that high plasma glucose and osmolality and high renal tubular glucose and osmolality are somehow involving in renal NOS synthesis in diabetic rats. Phlorizin, a competitive inhibitor of glucose transport in the proximal tubule, causes renal glycosuria in nondiabetic rats and reverses hyperglycemia in diabetic rats. To further investigate whether high plasma glucose and osmolality or high renal tubular glucose and osmolality influence renal NOS synthesis in diabetic rats, we measured nNOS mRNA levels in phlorizin treated normal and diabetic rats. Neuronal NOS mRNA expression in the kidneys was not significantly different between normal rats and phlorizin-treated normal rats with high urinary glucose and osmolality. The phlorizin-treated diabetic rats showed a significant decrease in the ratio of nNOS to beta-actin mRNA compared with diabetic rats. On linear-regression analysis, plasma glucose was strongly positively correlated with nNOS mRNA expression in the cortex, outer medulla, and inner medulla (r(2) =.378, r(2) =.680, and r(2) =.445, respectively) of rat kidneys. Neither urine glucose concentration nor urine osmolality was correlated with nNOS mRNA expression in rat kidneys. In conclusion, our results indicate that nNOS mRNA expression in the kidneys of diabetic rats is directly affected by high blood glucose/osmolality but not by high urinary glucose or osmolality. PMID- 12624602 TI - Tetrathiomolybdate therapy protects against bleomycin-induced pulmonary fibrosis in mice. AB - Tetrathiomolybdate (TM), a drug developed for the treatment of Wilson's disease, produces an antiangiogenic effect by reducing systemic copper levels. Several angiogenic cytokines appear to depend on normal levels of copper for activity. In both animal tumor models and in cancer patients, TM therapy has proved effective in inhibiting the growth of tumors. We have hypothesized that the activities of fibrotic and inflammatory cytokines are also subject to modulation by the availability of copper in a manner similar to angiogenic cytokines. As a first step in evaluating whether TM plays a therapeutic role in diseases of inflammation and fibrosis, we studied the effects of TM on a murine model of bleomycin-induced pulmonary fibrosis. Oral TM therapy resulted in dose-dependent reduction in serum ceruloplasmin, a surrogate marker of systemic copper levels. Significant decreases in systemic copper levels were associated with marked reduction in lung fibrosis as determined on the basis of histopathologic findings and a biochemical measure of fibrosis. The protection afforded by TM was also reflected in significantly reduced bleomycin-induced body-weight loss. In the next phase of this work, we will seek to determine the mechanisms by which TM brings about this therapeutic benefit. PMID- 12624603 TI - A novel early chorioallantoic membrane assay demonstrates quantitative and qualitative changes caused by antiangiogenic substances. AB - The chicken chorioallantoic membrane (CAM) has been extensively used in the study of angiogenesis. However, the CAM assay can be difficult and time-consuming to quantify, provides poor quality images of the results, and is not very reproducible. In this study, a novel early CAM assay was developed: It was found to be quantitative through relatively simple methods, enabled high-quality imaging of results, and was reproducible. Additionally, unique qualitative changes in vessel structure were observed, and it was possible to measure veins and arteries separately. Treatment of the CAM on days 4 and 5 with SU5614, suramin, fumagillin, amiloride, and PI-88 reduced blood-vessel growth. SU5614 (4 microg) resulted in significant reductions in artery but not vein length (60% and 111%, respectively, vs control). Suramin tended to increase CAM vasculature at 50 microg but caused dramatic reductions both in vessel length and CAM growth at 100 microg. As with SU5614, the effect was greater with regard to arterial compared with venous length (49% and 74%, respectively, vs control). PI-88 (20 microg) also decreased artery and vein length (66% and 80%, respectively, vs control). In contrast, fumagillin (5 microg) and amiloride (20 microg) both reduced arterial growth slightly less than venous growth (67% and 54% and 50% and 44%, respectively, vs control). Each antiangiogenic substance caused a different qualitative pattern of change in vessel branching and structure. The early CAM assay will be useful in the screening of antiangiogenic substances. Further study of the qualitative effects of antiangiogenic treatments may be a valuable tool to increase our understanding of the angiogenic process itself. PMID- 12624604 TI - Diastolic blood pressure is an important determinant of augmentation index and pulse wave velocity in young, healthy males. AB - Pulse wave velocity (PWV) and augmentation index are widely used measures of arterial stiffness. The purpose of this study was to evaluate the role of blood pressure as a determinant of both indices independent of potentially confounding factors including gender, age and cardiovascular disorders. A total of 77 young, healthy subjects were investigated under resting conditions. Augmentation index was derived by pulse wave analysis using carotid applanation tonometry. PWV was determined from pressure tracing over the carotid and femoral artery. The relations between stiffness markers and haemodynamic parameters were analysed by simple (r) and multiple (beta) regression analysis. Using simple regression analysis, augmentation index was correlated to age (r=0.292, P=0.0105), diastolic blood pressure (DBP, r=0.483, P<0.0001), mean arterial blood pressure (MAP, r=0.381, P=0.0007), pulse pressure (r=-0.414, P=0.0002) and total peripheral resistance (r=0.266, P=0.0204). After multiple regression analysis, augmentation index remained significantly correlated only to DBP (beta=0.347, P=0.0051). Using simple regression analysis, PWV was correlated to age (r=0.304, P=0.0067), systolic blood pressure (r=0.280, P=0.0129). DBP (r=0.455, P<0.0001), MAP (r=0.446, P&<0.0001) and heart rate (r=0.348, P=0.0018). After multiple regression analysis, PWV remained correlated only to age (beta=0.218, P=0.0422) and DBP (beta=0.4105, P=0.0316). In summary, DBP is an important determinant of augmentation index and PWV in young, healthy males. Further studies are needed to characterize the impact of blood pressure on arterial stiffness in other populations including females and older subjects. PMID- 12624605 TI - The determinants of left ventricular hypertrophy defined by Sokolow-Lyon criteria in untreated hypertensive patients. AB - Left ventricular hypertrophy (LVH) measured by electrocardiography (ECG LVH) in hypertensive patients has been shown to be associated with an increased risk of cardiovascular sequelae. Analysis of the determinants predisposing to ECG LVH may be helpful in the prevention of LVH. The Department of Health and Social Security Hypertension Care Computer Project studied 2994 hypertensive patients in whom an electrocardiogram was recorded while not on treatment. LVH was determined as the voltage sum SV1+RV5 or RV6>or=35 mm using Sokolow-Lyon voltage criteria. The relations were determined between the presence of LVH or voltage sum and different variables. Untreated systolic (SBP) and diastolic (DBP) blood pressure and pulse pressure were positively related to the increasing ECG voltage, while body mass index (BMI) and serum cholesterol were inversely related. Blood glucose and age did not correlate significantly. Patients with the presence of ECG LVH were more often men, black people, smokers and users of alcohol. In multiple logistic regression analyses, SBP, DBP, male gender and black race were positively, whereas BMI was negatively related to the presence of LVH. The positive relation of smoking and negative relation of serum cholesterol concentration to the presence of ECG LVH were apparent in men but not in women. This study confirms the adverse association between ECG LVH and SBP and DBP, male gender, black race and decreased BMI. It also addresses the less well-known associations of blood glucose, cholesterol, smoking and alcohol consumption. PMID- 12624606 TI - Circulating homocysteine levels in sustained and white coat hypertension. AB - Although white coat hypertension has been widely studied in the last years, its risk profile is not yet completely clear. The aim of this study was to evaluate circulating homocysteine levels, an emerging cardiovascular risk factor, in subjects with white coat and sustained hypertension. We selected 31 sustained hypertensive subjects, 31 white coat hypertensive subjects and 31 normotensive subjects matched for age, gender, body mass index and occupation. Women were also matched for menopausal status. Subjects with smoking habit, dyslipidaemia and diabetes mellitus were excluded from the study. White coat hypertension was defined as clinical hypertension and daytime ambulatory blood pressure <135/85 mmHg. Blood samples were drawn after a fasting period of 12 h for routine laboratory tests and homocysteine determination. Homocysteine levels were evaluated by fluorescence polarization immunoassay. Creatinine, glucose, cholesterol and triglycerides were not different among the groups. White coat hypertensive subjects had significantly lower homocysteine levels than sustained hypertensive patients (8.2+/-2.0 vs 12.6+/-3.9 micromol/l, P=0.0003). No significant difference was observed between white coat hypertensive and normotensive subjects regarding this parameter (8.2+/-2.0 vs 7.6+/-1.9 micromol/l, P=0.9). In conclusion, our data show that middle-aged white coat hypertensive subjects without other cardiovascular risk factors have lower circulating homocysteine levels than sustained hypertensive patients suggesting that they are at lower cardiovascular risk. PMID- 12624607 TI - Reduced heart rate variability in hypertension: associations with lifestyle factors and plasma renin activity. AB - Limited information exists on the relations between heart rate variability, hypertension, lifestyle factors and renin-angiotensin-aldosterone system. A total of 191 newly diagnosed yet untreated hypertensive men and women, 35-54 years of age, were compared with an age- and gender-stratified random population sample of 105 normotensive men and women to find out independent determinants of heart rate variability. Heart rate variability was computed from 5-min ECG time series using the standard deviation of normal-to-normal RR intervals (SDNN), the square root of the mean of squared differences between adjacent normal RR intervals (RMSSD) and the fast Fourier transform spectral analysis. All absolute measures of heart rate variability were reduced in hypertension (P<0.001 for each, ANOVA). In multivariate regression analyses, reduced heart rate variability was independently associated with higher heart rate (P<0.001 for all absolute measures of heart rate variability), higher age (P=0.001 for SDNN, total and LF powers; P<0.001 for RMSSD and HF power) and higher mean arterial pressure (P<0.05 for total power, P<0.01 for the other absolute measures) but not with sodium and alcohol intakes, body mass index and smoking. Increased plasma renin activity (PRA) was an independent attributor of reduced HF power (P<0.05) and reduced RMSSD (P<0.01). Increased blood pressure and heart rate are associated with decreased heart rate variability without any direct effects on heart rate variability of lifestyle factors. High PRA is an independent determinant of diminished modulation of vagal activity. PMID- 12624608 TI - Blood pressure reactivity to mental stress task as a determinant of sustained hypertension after 5 years of follow-up. AB - Previous studies have reported an increased risk of developing sustained hypertension (SH) in borderline or mildly hypertensive subjects showing an exaggerated response of blood pressure (BP) to mental stress. The aim of this study was to assess if the response of BP to mental stress tasks is an independent predictor of SH. A total of 89 patients with grade 1 hypertension, aged 18-64 years, 62% males, were included. The mean of follow-up was 5.3 years (s.d. 2.1 years). SH was defined as the development of grades 2-3 hypertension (Systolic BP>or=160 mmHg or diastolic BP>or=100 mmHg) or to be in antihypertensive treatment after follow-up. Two mental stress tasks: mental arithmetic stress task and a stressful interview (SI) were applied at entry. The subjects were classified as hyper-reactors when BP increase was greater than 35 mmHg for systolic BP or greater than 21 mmHg for diastolic BP, according to the results obtained previously in a normotensive control group. In the univariate analysis, the factors associated with the development of SH were age (P=0.0007), office diastolic BP (P=0.014) and hyper-reactivity of BP during a stressful interview (P=0.003). In the Cox regression model, after adjusting for gender, age, and office BP, the hyper-reactivity of BP during SI was an independent predictor of development of SH. In conclusion, the response of BP to mental stress tasks is useful in predicting SH in young and middle-aged subjects with grade 1 hypertension. PMID- 12624609 TI - Renin-angiotensin-aldosterone system and G-protein beta-3 subunit gene polymorphisms in salt-sensitive essential hypertension. AB - Approximately 50% of hypertensive patients are salt sensitive (they increase their Blood Pressure in response to sodium intake or volume expansion). Mechanisms underlying salt sensitivity are not completely elucidated although there is evidence that they may be genetically determined. The aim of this study is to establish the relation among some genetic polymorphisms of the renin angiotensin system (RAAS) and the beta-3 subunit of the protein G and salt sensitivity. We studied 102 essential hypertensive patients, stage 1-2 and without target organ damage. Salt sensitivity was assessed by the rapid protocol of Weinberger. We determined by polymerase Chain reaction techniques the following polymorphisms: insertion/deletion (I/D) of the angiotensin-converting enzyme (ACE), A1166C of the angiotensin II type 1 receptor (AT1R), -344C/T and intron 2 conversion (IC) of the aldosterone synthase (CYP11B2), and C825T of the beta-3 subunit of the protein G (GNB3). 41 patients (40.19%) were salt sensitive. The distribution of the different polymorphisms was similar in both groups of patients, but subjects carriers of the W allele of the CYP11B2 IC polymorphism had a greater risk for salt sensitivity as compared with no carriers (37 of 41, 90.2% vs 4 of 41, 9.8%, OR 3.02, P<0.05). Although there is no association between salt sensitivity and the different studied genotypes of the RAAS and of the GNB3, our data show a greater risk for salt sensitivity among carriers of the W allele of the CYP11B2 1C polymorphism. PMID- 12624610 TI - Sagittal abdominal diameter is associated with insulin sensitivity in Chinese hypertensive patients and their siblings. AB - The purpose of the study is to observe the relation between anthropometric measurements, focusing on sagittal abdominal diameter (SAD), and insulin sensitivity indices in Chinese hypertensive patients and their siblings. In total, 907 participants, 537 hypertensive and 370 nonhypertensive, from 311 Taiwanese families were drawn from the Stanford Asia and Pacific Program for Hypertension and Insulin Resistance for the study. The participants received anthropometric measurements and 75-g oral glucose tolerance tests after an overnight fast. Fasting insulin, homeostasis model assessment for insulin resistance (HOMA-IR), and the insulin sensitivity index ISI(0,120) were chosen as surrogate measures of insulin sensitivity. In addition to Pearson and partial correlations, we used generalized estimating equations (GEEs) to examine the association between anthropometric measurements and insulin sensitivity indices. A small deviance in the GEEs indicates the goodness of model fit, irrespective of the independence among variables. The hypertensive patients were older in age, wider in waist circumference (WC), larger in body mass index (BMI) and SAD, and more insulin resistant than the nonhypertensive counterparts. The logarithmic transformation of fasting insulin, HOMA-IR, and ISI(0,120) significantly correlated with SAD, WC, and BMI before and after adjustments for age and sex. The deviances of SAD in the GEEs were similar to those of WC in all subjects, while BMI had smaller deviances than SAD and WC in the hypertensive patients. Our results suggest that the performance of SAD in predicting insulin sensitivity is comparable with WC in Chinese hypertensive patients and their siblings. BMI, however, seems to have better association with insulin sensitivity than SAD and WC in the patients with hypertension. PMID- 12624611 TI - Effectiveness of multidisciplinary lifestyle intervention for hypertension: a randomised controlled trial. AB - Lifestyle factors like weight, alcohol consumption, salt intake and physical activity have shown to be important in treating hypertension. There have been made some randomised trials about the effects of lifestyle interventions, but the numbers of patients have been relatively small and the durations of follow-ups have been short. No controlled trials assessing the effects of lifestyle intervention in a rehabilitation setting have been reported. In this study, the effects of multidisciplinary lifestyle intervention in rehabilitation centres among middle-aged hypertensive employees were described. A total of 731 hypertensives from 45 worksites were randomised to lifestyle intervention in a rehabilitation centre or to usual care in an occupational or primary health-care centre for 12 months. Standard measurements were conducted before the intervention and 1-year later. Blood pressure (BP) levels were clearly reduced in the intervention group, while only minor changes were observed in the control group. The net changes between the two groups both for systolic and diastolic BPs were -2.1 mmHg (95% confidence intervals (CI) -4.0 to -0.1) and -1.5 mmHg (95% CI -2.6 to -0.4), respectively. The net changes were greater among men than women. The multidisciplinary lifestyle intervention in a rehabilitation centre setting produced significant reductions in BP among middle-aged employees with hypertension. PMID- 12624613 TI - Stenting of a renal artery compressed by the diaphragm. AB - A 74-year-old man had a resistant hypertension with an increase in plasma aldosterone and active plasma renin levels, and an irregular appearance of the left kidney outline by ultrasound. The CT scan showed a stenosis of the left renal artery, which was pushed against the aorta by the left crus of the diaphragm. An angioplasty with placement of an autoexpansible stent was carried out with a good result on the arterial pressure level. After 3 years, the patient was re-hospitalised with severe hypertension. The CT scan demonstrated a compression of the stent by the left crus of the diaphragm, with good permeability of the artery downstream from the stent, and radiographic examination showed a fracture of the left renal artery stent. Thus, a reimplantation of the left renal artery in the aorta was carried out. Stenosis of the renal artery by fibres from a crus of the diaphragm is a rare cause of renovascular hypertension. Helicoidal angioscanner imaging is particularly useful to do the diagnosis. In the present case, renal angioplasty with stenting was complicated by a fracture of the stent that led to the surgery. Thus, when renal artery stenosis by a crus of the diaphragm is diagnosed, surgical treatment needs to be considered on a case-by-case basis in relation to the anatomy and the biological and functional data. PMID- 12624612 TI - Effect of benazepril addition to amlodipine on ankle oedema and subcutaneous tissue pressure in hypertensive patients. AB - The aim of this study was to evaluate the effect of benazepril addition to amlodipine antihypertensive treatment on ankle-foot volume (AFV) and pretibial subcutaneous tissue pressure (PSTP), two objective measures of ankle oedema. A total of 32 mild to moderate essential hypertensives (DBP>90 and <110 mmHg), aged 30-70 years were studied. After a 4-week placebo period, they were randomized to amlodipine 5 mg o.d. or benazepril 10 mg o.d. or amlodipine 5 mg plus benazepril 10 mg o.d. for 4 weeks, according to a crossover design. At the end of the placebo period and of each active treatment period, blood pressure,AFV and PSTP were evaluated. AFV was measured using the principle of water displacement. PSTP was assessed using a system, the subcutaneous pretibial interstitial environment with a water manometer. Both amlodipine and benazepril monotherapy significantly reduced SBP (-18.2+/-4 and -17.8+/-4 mmHg, respectively, P<0.01 vs baseline) and DBP (-12.1+/-3 and -11.7+/-3 mmHg, respectively, P<0.01); the reduction was increased by the combination (-24.2+/-5 mmHg for SBP, P<0.001 and -16.8+/-4 mmHg for DBP, P<0.001). Amlodipine monotherapy significantly increased both AFV (+17.1%, P<0.001 vs baseline) and PSTP (+56.6%, P<0.001 vs baseline). As compared to amlodipine alone, the combination produced a less pronounced increase in AFV (+5.5%, P<0.05 vs baseline and P<0.01 vs amlodipine) and PSTP (+20.5%, P<0.05 vs baseline and P<0.01 vs amlodipine). Ankle oedema was clinically evident in 11 patients with amlodipine monotherapy and in three patients with the combination. These results suggest that ACE-inhibitors partially counteract the microcirculatory changes responsible for Ca-antagonists-induced oedema formation. PMID- 12624614 TI - Determinants of poor hypertension management in the community. PMID- 12624615 TI - End points in asthma. PMID- 12624617 TI - Spirometry and tuberculosis in Ecuador. PMID- 12624618 TI - The Kyoto Accord and the medical profession. PMID- 12624619 TI - Clinical models to compare the safety and efficacy of inhaled corticosteroids in patients with asthma. AB - There is no consensus on the methods to compare the clinical efficacy of different inhaled corticosteroids. A comparison needs to be made in terms of relative potency, and studies should include two-, or preferably, three-dose comparisons. A number of clinical models and outcomes are available; they have their relative advantages and disadvantages. While measurements of symptoms and spirometry are easy and readily available, they show a flat dose-response relationship. Measurements of bronchial hyper-responsiveness to exercise and adenosine monophosphate, allergen-induced airway responses, and measurements of inflammation in sputum and exhaled air show steep dose-response relationships, particularly to low doses of inhaled steroids. An uncontrolled asthma model followed by stabilization with a short course of additional steroid, with measurements of airway responsiveness and airway inflammation, in a crossover study seems more promising than the other models. Drug deposition studies and mathematical modelling of drug pharmacokinetics in the airway may provide complementary information to clinical drug relative potency studies. Fine particle dose and emitted doses, rather than the nominal dose, should be considered in the estimation of clinical and systemic effects, respectively. When a second entry (generic) drug is being evaluated in comparison with the innovator drug (same compound and same device), it may be appropriate to consider accepting a generic as bioequivalent if it satisfies pharmaceutical equivalence. PMID- 12624620 TI - Synchronous lung and liver metastases from medullary thyroid carcinoma. AB - Metastatic disease is one of the most common causes of calcified nodules in the lung or liver. The incidence of calcified metastasis mainly to the lung and liver is high at the initial presentation in patients with medullary thyroid carcinoma. Synchronous calcified metastasis in the lung and liver is reported for the first time. The diagnosis of medullary thyroid carcinoma may be evident from the synchronous presence of miliary calcified nodules in two different sites if they are associated with high concentrations of serum markers. PMID- 12624621 TI - Sildenafil treatment of primary pulmonary hypertension. AB - A 37-year-old woman with primary pulmonary hypertension and worsening symptomatology underwent pulmonary artery (PA) catheterization and vasodilator trials. Oxygen had no effect, but 10 parts/million of nitric oxide reduced mean PA (PAm) pressure by 20%. Prostacyclin infusion at 8 ng/kg/min decreased the PAm pressure by 11%, but further dose increases were limited by systemic hypotension. Sildenafil in doses of 25 mg or higher resulted in an average decrease of 14% in PAm pressure. Sildenafil is a potentially useful treatment option for patients with primary pulmonary hypertension, and further investigation is warranted. PMID- 12624622 TI - Molecular mechanisms of initiation of fibrinolysis by fibrin. AB - Fibrinogen is rather inert in the circulation, however, after conversion into fibrin it participates in various physiological processes including fibrinolysis. Initiation of fibrinolysis occurs through a number of orchestrated interactions between fibrin, plasminogen and its activator tPA which result in generation of plasmin. Numerous studies localized a set of specific low affinity tPA- and plasminogen-binding sites in each D region of fibrin(ogen). The tPA-binding site includes residues gamma312-324 and the plasminogen-binding site includes residues Aalpha148-160; they bind tPA and plasminogen with a K(d) of about 1 micro M. Another set of high affinity tPA- and plasminogen-binding sites (K(d)s = 16-33 nM) was identified in the compact portion of each fibrin(ogen) alphaC-domain within residues Aalpha392-610. All these sites are cryptic in fibrinogen and become exposed in fibrin. Recent studies with recombinant and proteolytic fibrin(ogen) fragments clarified the molecular mechanisms by which these sites become exposed. Namely, upon fibrin assembly, the interaction between the D and E regions causes conformational changes in the former that expose the low affinity binding sites. The exposure of the high affinity binding sites in the alphaC domains is connected most probably with their switch from an intramolecular interaction in fibrinogen to an intermolecular one in fibrin. These mechanisms serve to minimize degradation of circulating fibrinogen and confine fibrinolysis to places of fibrin deposition. PMID- 12624623 TI - Molecular evolution of the vertebrate blood coagulation network. AB - In mammalian blood coagulation 5 proteases, factor VII (FVII), factor IX (FIX), factor X (FX), protein C (PC) and prothrombin act with two cofactors factor V and factor VIII to control the generation of fibrin. Biochemical evidence and molecular cloning data have previously indicated that blood coagulation involving tissue factor, prothrombin and fibrinogen is present in all vertebrates. Using degenerate RT-PCR we have isolated and characterized novel cDNAs with sequence identity to the blood coagulation serine proteases and cofactors from chicken and the puffer fish (Fugu rubripes). Sequence alignments, phylogenetic and comparative sequence analysis all support the existence of the Gla-EGF1-EGF2-SP domain serine proteases FVII, FIX, FX, PC and the A1-A2-B-A3-C1-C2 domain protein cofactors FV and FVIII in these species. These results strongly suggest that the blood coagulation network is present in all jawed vertebrates and evolved before the divergence of tetrapods and teleosts over 430 million years ago; and that vertebrate blood coagulation may have benefited from two rounds of gene or whole genome duplication. Sequences identified in Fugu coding for additional FVII-like, FIX-like and PC-like sequences support the possibility of further tandem and large-scale duplications in teleosts. Comparative sequence analyses of amino acid residues in the active site region suggest these additional sequences have evolved new and as yet unknown functions. PMID- 12624624 TI - The R2-haplotype associated Asp2194Gly mutation in the light chain of human factor V results in lower expression levels of FV, but has no influence on the glycosylation of Asn2181. AB - The R2 haplotype of the FV gene spans from exon 8 through 25 and comprises several strongly linked polymorphisms in the FV gene, including some missense mutations. Carriership of the R2-FV allele has been associated with reduced plasma FV levels, increased FV1/FV2 ratios and mild APC resistance. Some studies have reported that carriership of the R2-FV allele is associated with an increased risk of venous thombosis. At this moment, the individual contribution to the R2-associated phenotypes of the different mutations linked to the R2 haplotype of FV is unclear. The main objective of our study was to obtain insight in the influence of the R2-related Asp2194Gly mutation on FV expression, FV structure and FV function using Bdomainless rFV mutants. Replacing Asp at position 2194 by Gly resulted in a more than threefold reduction of rFV expression compared to rFV wild-type. Therefore, we propose that the R2-linked Asp2194Gly mutation is an important determinant of the association of the R2-FV allele with lower FV levels. Furthermore, the light chains from Asp2194Gly containing rFV mutants showed similar molecular weights as the light chains of the non-glycosylated rFVwt or the plasma FV2 isoform, indicating that glycosylation at Asn2181 is not stimulated by the presence of a glycine in position 2194. Finally, the apparent K(d) for dissociation of the FXaVa complex (K(1/2Xa)) was not higher in rFV mutants with the Asp2194Gly mutation than for rFVwt, suggesting that also the affinity for negatively charged phospholipids is not affected by substitution of Asp into Gly at position at 2194. PMID- 12624625 TI - Local tissue factor pathway inhibitor release in the human forearm. AB - Nineteen healthy men received unilateral brachial artery infusions of either unfractioned heparin (0.3-100 IU/min), saline or the endothelium-dependent vasodilators substance P (2-8 pmol/min) and bradykinin (100-1000 pmol/min), and the endothelium-independent vasodilator sodium nitroprusside (2-8 micro g/min). Heparin caused a dose-dependent increase in plasma TFPI concentrations in both arms (ANOVA, p <0.0001). Estimated net forearm TFPI release was 7 +/- 16, 29 +/- 20 and 138 +/- 72 ng/100 mL tissue/min during 10, 30 and 100 IU/min of heparin respectively (ANOVA, p <0.0001). Compared to the systemic circulation, the forearm sensitivity to heparin induced TFPI release was 3.6-fold lower (166 +/- 67 ng/IU vs. 596 +/- 252 ng/IU: t-test, p = 0.004). Substance P, bradykinin and sodium nitroprusside all caused substantial dose-dependent increases in blood flow (ANOVA, p <0.001 for all) without affecting plasma TFPI concentrations. There are important regional differences in endothelial TFPI release, with the forearm circulation being relatively insensitive to heparin. PMID- 12624626 TI - No association between thrombosis and factor V gene polymorphisms in Chinese Han population. AB - Activated protein C resistance (APCR) is the most common hereditary condition of thrombosis in Western countries. And it is significantly linked to a single nucleotide polymorphisms (SNPs) in the coagulation factor V gene that results in the mutations at R506, R306 and HR2 alleles. To determine the prevalence of APCR and its association with the factor V gene SNPs in Chinese Han thrombotic patients, we investigated a total of 346 Chinese thrombotic patients and 140 normal controls for APCR using the APTT-based assays, according to manufacturer's instructions, APC ratio /=30 kg/m(2)) increased the risk of thrombosis twofold (CI95: 1.5 to 3.4), adjusted for age and sex. Obese individuals had higher levels of factor VIII and factor IX, but not of fibrinogen. The effect on risk of obesity was not changed after adjustment for coagulation factors levels (fibrinogen, F VIII, F IX, D-dimer). The relative risk estimates were similar in different age groups and in both sexes, indicating a larger absolute effect in older age groups. Evaluation of the combined effect of obesity and oral contraceptive pills among women aged 15-45 revealed that oral contraceptives further increased the effect of obesity on the risk of thrombosis, leading to 10-fold increased risk amongst women with a BMI greater than 25 kg/m(2) who used oral contraceptives. Obesity is a risk factor for deep vein thrombosis. Among women with a BMI greater than 25 kg/m(2) the synergistic effect with oral contraceptives should be considered when prescribing these. PMID- 12624634 TI - Travel and the risk of symptomatic venous thromboembolism. AB - Whether long-distance travel and symptomatic venous thromboembolism (VTE) are associated is debated. On the basis of the available literature a fair risk estimate cannot be obtained. We estimated an accurate odds ratio for the relationship between recent travelling and symptomatic VTE. From three case control studies consisting of 788 and 170 patients with clinically suspected deep vein thrombosis (DVT) and 989 patients with clinically suspected pulmonary embolism (PE) referred for diagnostic work-up, a pooled odds ratio for the relation between travel and symptomatic VTE was calculated. Cases were patients in whom the diagnosis was confirmed according to a diagnostic management strategy, whereas controls were patients in whom the diagnosis was excluded and who had an uneventful clinical follow-up. Patients were seen in the period April 1997 to September 2000. Travel history was recorded prior to diagnostic work-up. The pooled odds ratio for the association between any travel and symptomatic venous thromboembolism was 0.9 (95% CI: 0.6-1.4). The median travel time was 7 h (quartile range 4 to 10 h). Separate analyses performed for different types of transport (plane, car, bus or train) yielded comparable odds ratios. The analysis for duration of travelling showed an increased odds ratio of 2.5 (95% CI: 1.0 6.2) in the category of 10-15 h of travelling. This study shows that the average traveller does not have an increased risk for symptomatic venous thromboembolism. Only very long travelling (more than 10 h) may be associated with venous thromboembolic disease. PMID- 12624635 TI - Enzyme immunoassay detection of platelet-derived microparticles and RANTES in acute coronary syndrome. AB - Platelet-derived microparticles (PDMPs) are produced by platelet activation or physical stimulation under various conditions. To evaluate changes in platelet and chemokine function in patients undergoing percutaneous transluminal coronary angioplasty (PTCA), we measured and compared levels of PDMPs and a C-C chemokine, regulated on activation normally T-cell express and secreted (RANTES), by ELISA. Levels of PDMP and RANTES in patients with acute coronary syndrome were significantly higher than those in the control groups (PDMP: 20.1 +/- 2.9 vs 80.4 +/- 7.3 U/ml, p < 0.001; RANTES: 18.6 +/- 3.7 vs 52.1 +/- 4.6 ng/ml, p < 0.01), but did not differ between the control groups and patients with stable angina. PDMP levels were higher in patients with acute myocardial infarction (AMI) than in patients with unstable angina (PDMP: 115.0 +/- 7.1 vs 63.9 +/- 6.2 U/ml, p < 0.001). There was no difference in the RANTES levels, however, between patients with AMI and patients with unstable angina. PDMP and RANTES levels were significantly decreased after PTCA (PDMP, p < 0.001; RANTES, p < 0.05), but without differences between the two groups. In addition, the level of PDMP was significantly correlated with that of RANTES or soluble CD40 ligand. These findings suggest that PTCA may prevent the development of AMI-associated complications in which activated platelets and RANTES play roles. Our ELISA method appears to be sufficient for monitoring PDMP and RANTES levels after PTCA in patients with acute coronary syndrome. PMID- 12624636 TI - CD32-dependent platelet activation by a drug-dependent antibody to glycoprotein IIb/IIIa antagonists. AB - Thrombocytopenia is observed with a frequency of up to 2% in patients treated with glycoprotein (GP) IIb/IIIa antagonists. We recently provided evidence that thrombocytopenia is caused by antibody binding to drug-induced conformational changes in GP IIb/IIIa. Here, we report that a murine monoclonal antibody binds to GP IIb/IIIa in an antagonist-dependent manner and activates platelets. Platelet stimulation is associated with a disruption of the phospholipid asymmetry, resulting in the assembly of catalytic active intrinsic Xase and prothrombinase complexes. Further mechanistic studies revealed that this response is (I) mediated in cis, (II) not associated with the formation of prothrombotic microparticles, and (III) requires intact platelet signaling and (IV) is blocked by increases in cAMP. The prothrombotic response is not observed using F(ab')2 fragments and is blocked by incubation of platelets with neutralizing antibodies to the platelet FcgammaRIIa receptor (CD 32).Taken together, these observations suggest that GPIIb/IIIa antagonist-dependent antibody binding to the platelet fibrinogen receptor has the propensity to lead to CD32-mediated platelet activation and accelerated platelet clearance, leading to thrombocytopenia. PMID- 12624637 TI - Focusing on transcription factor families in atherogenesis: the function of LKLF and TR3. AB - In this overview, two separate studies are discussed that emerged from a "discovery-driven" approach to identify genes that play an essential role in atherogenesis. First, by a combination of DNA micro-array and one-way linkage hierachical clustering, we selected genes that are induced in endothelial cells (EC) by prolonged steady- or pulsatile laminar flow, but of which expression is not affected by inflammatory and mitogenic agents (TGF-beta, IL-1betaTNF alpha,VEGF, thrombin). The genes selected accordingly were: cytochrome P450 1B1, diaphorase and the transcription factor lung Kruppel-like factor (LKLF) of which only the latter is truly EC specific. LKLF meets the criteria of an anti atherosclerotic gene, mainly since expression is restricted to areas subjected to laminar flow as shown by in situ hybridization with anatomically well-defined specimens. Second, neointimal (but not medial) smooth muscle cells (SMC) specifically synthesize the NGFI-B subfamily (TR3, MINOR,NOT) of the nuclear hormone superfamily of transcription factors. Again, evidence is presented, indicating that these proteins serve an anti-atherosclerotic function. Notably, transgenic mice, expressing either TR3 or a dominant-negative mutant TR3DeltaTA in arterial SMC, were subjected to carotid artery ligation to induce SMC proliferation. Lesions in TR3-overexpressing transgenic mice were 5-fold smaller than isogenic wild-type mice, while mice overexpressing the TR3DeltaTA mutant had a 3-fold larger lesion. It is proposed that (down-stream products of) TR3 inhibit the cell cycle, since adenovirus-mediated expression of TR3DeltaTA and TR3, respectively, inhibit and promote the synthesis of the cyclin-dependent kinase inhibitor p27(Kip1) in SMC. PMID- 12624639 TI - Rise of circulating thrombopoietin following cardiothoracic surgery is potentiated in patients with coronary atherosclerosis: correlation with a preceding increase in levels of interleukin-6. AB - Thrombopoietin (TPO) is the major regulator of platelet production. Plasma levels of TPO are thought to be regulated by its binding to platelets and megakaryocytes. Here we have used a model of cardiac surgery with cardiopulmonary bypass (CBP) to test the possibility that changes in TPO levels are influenced by the presence of coronary artery disease (CAD) and by changes in interleukin-6 (IL 6). After surgery patients with CAD (n = 22) or with normal coronary arteries (n = 11) showed a significant thrombocytopaenia followed by a reactive thrombocytosis. The platelet recovery was preceded by a significant rise in TPO (from 62.6 +/- 9.4 pg/ml at baseline to 129.2 +/- 19 pg/ml at 60 h, P <0.001), which in turn was preceded by, and was positively correlated with, a marked increase in circulating IL-6 (from 1.5 +/- 0.3 pg/ml at baseline to 269.3 +/- 30.6 pg/ml at 3-12 h, P <0.001). The rise of both IL-6 and TPO was significantly larger in patients with CAD. No correlation was found between the post-operative drop in platelet mass and changes in either the TPO or IL-6 levels. These findings suggest that in man circulating TPO levels, besides being controlled by changes in platelet mass, are influenced by inflammatory processes, including the presence of coronary atherosclerosis. PMID- 12624638 TI - Insights in the molecular mechanisms of the anti-angiogenic effect of an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase. AB - 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) reduce the risk of coronary event by cholesterollowering dependent and independent mechanisms. We have already described that the inhibitory effect of cerivastatin on angiogenesis contribute to the cholesterol-independent beneficial effect and was due to the inhibition of the cell signaling cascade RhoA/FAK/Akt. In this study, new insights in the molecular mechanism of action were provided. It indicates an inhibition of exposure of alpha V beta 3 integrin on cell membrane and a modification of gene expression. The inhibition of angiogenesis could be related to 1) an increase in genes involved in the inhibition of cell proliferation (p19(INK4), p21(Waf/Cip1),Wnt-5a), the inhibition of cell migration (Rho-GDI 1, alpha E-catenin) and 2) a downregulation of genes involved in angiogenesis (PAI-1, Vitronectin, HoxD3, Notch4) or in cell invasion (Semaphorin E). In addition, DNA repair protein genes (MLH1, XRCC1) were increased. This study may indicate new biological interest of genes involved in angiogenesis control. PMID- 12624640 TI - Vascular thrombogenicity induced by progressive LDL oxidation: protection by antioxidants. AB - Oxidative modification of LDL, which dysregulates the homeostasis between blood and vascular cells, and alterations of endothelial function are considered among the early events in the pathogenesis of atherosclerosis. This study was designed to evaluate the impact of progressive LDL oxidation on the thrombotic response both in vitro and in vivo, and to address the potential effect of antioxidants. Tissue factor was induced by progressive LDL oxidation in HUVEC, and this event was in parallel to the appearance of the apoptotic phenotype. Both these phenomena were mediated by ERK1/2 activation and were prevented by LDL pre enrichment with antioxidants. In contrast, antioxidants failed to affect tPA and PAI-1 secretion, which was increased by LDL, either native or oxidized. Tissue factor-pathway inhibitor was also increased upon HUVEC exposure to progressively oxidized LDL. LDL, in the presence of an oxidative agent, trigger a thrombogenic response in vivo, mostly TF-dependent, in an in situ model of platelet deposition. This effect was markedly attenuated when LDL were enriched with antioxidants. It can be concluded that vascular thrombogenicity is induced by progressive LDL oxidation and that alterations of the antioxidant/oxidant balance of the LDL particle in favor of the antioxidant tone are protective against the thrombotic response triggered by oxidative stress. The extrapolation of these data in a clinical setting, even if not easy, offers potential insights for the use of antioxidants in the prevention of thrombotic complications associated with atherothrombosis. PMID- 12624641 TI - Association between TAFI antigen and Ala147Thr polymorphism of the TAFI gene and the angina pectoris incidence. The PRIME Study (Prospective Epidemiological Study of MI). AB - Thrombin activatable fibrinolysis inhibitor (TAFI), a recently described inhibitor of fibrinolysis, has been hypothesized as playing a role in atherothrombosis. However, the evidence from retrospective studies, which have evaluated the role of TAFI in vascular risk, is conflicting. In a prospective cohort (the PRIME Study) of nearly 10 000 apparently healthy men recruited in France (Lille, Strasbourg, Toulouse) and Northern Ireland (Belfast), we measured baseline plasma concentration of TAFI antigen among 143 participants (81 from France and 62 from Ireland) who subsequently developed angina pectoris and among 286 age-matched participants who remained free of disease during the 5 years of follow-up. Genotyping of the Ala147Thr polymorphism located in the TAFI gene was performed using an allele specific PCR. In France, mean levels of TAFI were significantly higher at baseline among men who subsequently developed angina pectoris compared with their control subjects (119 versus 107 %; p = 0.02). The risk of future angina pectoris increased with increasing tertiles of TAFI (p = 0.02), such that men in the highest tertile at study entry had a 5-fold higher relative risk than those in the lowest tertile (95% confidence interval, 1.38 to 18.58) after controlling for the conventional cardiovascular risk factors. No such difference was observed in Northern Ireland. In France, Thr/Thr carriers of the Ala147Thr polymorphism were significantly more frequent in cases than in controls (p = 0.01) leading to a relative risk of angina pectoris of 2.7 (95%CI 1.2-5.8). Increase in plasma TAFI antigen levels is a risk factor for angina pectoris in France. Genotyping for the Ala147Thr polymorphism seems to be a reliable tool to assess the risk mediated by TAFI. PMID- 12624642 TI - Endothelial cell serine proteases expressed during vascular morphogenesis and angiogenesis. AB - Many serine proteases play important regulatory roles in complex biological systems, but only a few have been linked directly with capillary morphogenesis and angiogenesis. Here we provide evidence that serine protease activities, independent of the plasminogen activation cascade, are required for microvascular endothelial cell reorganization and capillary morphogenesis in vitro. A homology cloning approach targeting conserved motifs present in all serine proteases, was used to identify candidate serine proteases involved in these processes, and revealed 5 genes (acrosin, testisin, neurosin, PSP and neurotrypsin), none of which had been associated previously with expression in endothelial cells. A subsequent gene-specific RT-PCR screen for 22 serine proteases confirmed expression of these 5 genes and identified 7 additional serine protease genes expressed by human endothelial cells, urokinase-type plasminogen activator, protein C, TMPRSS2, hepsin, matriptase/MT-SP1, dipeptidylpeptidase IV, and seprase. Differences in serine protease gene expression between microvascular and human umbilical vein endothelial cells (HUVECs) were identified and several serine protease genes were found to be regulated by the nature of the substratum, ie. artificial basement membrane or fibrillar type I collagen. mRNA transcripts of several serine protease genes were associated with blood vessels in vivo by in situ hybridization of human tissue specimens. These data suggest a potential role for serine proteases, not previously associated with endothelium, in vascular function and angiogenesis. PMID- 12624643 TI - Fibrin-based biomaterials to deliver human growth factors. AB - Fibrin-based biomaterial preparations can be used as provisional growth matrices for cells important in tissue repair during wound healing in vivo. Their efficacy can be enhanced by including bioactive agents that promote specific cellular responses. This study examined the controlled delivery of the angiogenic growth factors bFGF, VEGF(165), and VEGF(121) using biomatrix preparations prepared from Fibrin Sealant product components. The growth factors were added prior to formation of the Fibrin Sealant clots, and the release kinetics of the proteins from the clots measured. The results indicated that the proteins were released from the clots more slowly in the order bFGF << VEGF(165) < VEGF(121). The biologic activity of the growth factors delivered from Fibrin Sealant clots was established by assaying growth stimulation of human microvascular endothelial cells (HMVEC) and angiogenesis in the chicken embryo chorioallantoic membrane (CAM) model of neovascularization. In the latter assay, clots containing bFGF, VEGF(165), or VEGF(121) all displayed angiogenic activity. However, delivery of either bFGF, VEGF(165), or VEGF(121) alone resulted in a significant percentage of clots becoming filled with blood, indicating that the newly developing vessels invading the clots were leaky and immature. In contrast, this hemorrhaging behavior did not occur with delivery of combinations, e.g., (VEGF(165) + VEGF(121)) or (VEGF(165) + bFGF), indicating that the vessels were more mature than those produced in response to single growth factors. Thus, delivering a combination of growth factors constituted an improvement over the delivery of individual growth factors for enhancing neovascularization. PMID- 12624644 TI - 2MHz ultrasound enhances t-PA-mediated thrombolysis: comparison of continuous versus pulsed ultrasound and standing versus travelling acoustic waves. AB - In addition to fibrinolytic enzymes, ultrasound has the potential to enhance thrombolysis. High frequency ultrasound has the advantage that a combination of diagnostic and therapeutic ultrasound with only one device is possible. Therefore, we investigated the optimal high frequency (2 MHz) ultrasound field characteristics and application mode in vitro. Continuous ultrasound significantly enhanced rt-PA mediated thrombolysis: in a travelling wave field thrombolysis was augmented by 49.0 +/- 14.7% and in a standing wave field by 34.8 +/- 7.3%. In an intermittent application mode (1Hz, 10Hz, 100Hz, 1kHz) most efficient results were obtained for both wave fields using 1 Hz (46.4 +/- 10.7% and 39.1 +/- 6.6%, respectively). Referring to a possible in vivo application our in vitro data suggests that an intermittent application of a 2 MHz high frequency ultrasound using a travelling wave field would be the most potent application for lysing blood clots. PMID- 12624645 TI - A safety analysis of thromboprophylaxis in acute medical illness. PMID- 12624646 TI - In silico tissue factor analysis: a bit-to-bit comparison. PMID- 12624647 TI - A reply to a rebuttal: the fibrinolytic system and obesity. PMID- 12624648 TI - The influence of climate on the basal metabolic rate of small mammals: a slow fast metabolic continuum. AB - The influence of climate (mean annual rainfall, rainfall variability, ambient temperature, T(a)) on the basal metabolic rate (BMR) of 267 small mammals (<1 kg) from six zoogeographical zones was investigated using conventional and phylogenetically independent data (linear contrasts). All climate variables varied between zones, as did BMR and body temperature ( T(b)), but not thermal conductance. Holarctic zones were more seasonal and colder, but rainfall was less variable, than non-Holarctic zones. In general, the BMR was most strongly influenced by body mass, followed by T(a) and the rainfall variables. However, there was significant variation in the strength of these relationships between zones. BMR and T(b) increased with latitude, and mass-independent BMR and T(b) were positively correlated. The latter relationship offers evidence of a slow fast metabolic continuum in small mammals. The fast end of the continuum (high BMR) is associated with the highest latitudes where BMR is most strongly influenced by T(a) and mean annual rainfall (i.e. mean productivity). The slow end of the continuum (low BMR) is associated with the semi-tropics, low productivity zones, and climatically unpredictable zones, such as deserts. Here rainfall variability has the strongest influence on BMR after body size. The implications of a slow-fast metabolic continuum are discussed in terms of various models associated with the evolution of BMR, such as the aerobic capacity models and the "energetic definition of fitness" models. PMID- 12624649 TI - Key metabolic enzymes and muscle structure in triplefin fishes (Tripterygiidae): a phylogenetic comparison. AB - Metabolic potential and muscle development were investigated relative to habitat and phylogeny in seven species of New Zealand triplefin fishes. Activity was measured in three principal glycolytic enzymes (lactate dehydrogenase, pyruvate kinase and phosphofructokinase) and two oxidative enzymes (citrate synthase and L3-hydroxyacyl CoA:NAD(+) oxidoreductase). The non-bicarbonate buffering capacity of caudal muscle was also estimated. Phylogenetic independent contrast analyses were used to reduce the effects of phylogenetic history in analyses. A positive relationship between metabolic potential and the effective water velocity at respective habitat depths was found only after the exclusion from analyses of the semi-pelagic species Obliquichthys maryannae. O. maryannae showed high glycolytic enzyme activities, and displayed double the activity of both oxidative enzymes relative to the six benthic species. Histochemically stained sections taken immediately posterior to the vent showed that adult O. maryannae and larval Forsterygion lapillum had significantly more red muscle, and smaller cross sectional areas of white and red muscle fibres, than adults of benthic species. The distribution of red muscle in adult O. maryannae resembled that of larval F. lapillum, and differed from the typical teleost pattern seen in adults of the six benthic species. Both adult O. maryannae and larval F. lapillum have an expansive lateralis superficialis muscle, typical of larval fish, which encompasses much of the caudal trunk. Results suggest that anaerobic potential in New Zealand triplefins: (a) increases with the locomotory requirements of different habitats, and (b) displays a negative relationship with depth-dependent water velocities in benthic species. O. maryannae appears to have increased aerobic potential for sustained swimming by paedomorphic retention of larval muscle architecture. PMID- 12624650 TI - Milk composition and growth in wild and captive Tasmanian bettongs, Bettongia gaimardi (Marsupialia). AB - Changes in milk composition (total solids, carbohydrate, protein, lipid and calculated gross energy content) during lactation in three groups of Tasmanian bettongs ( Bettongia gaimardi): free-living animals (wild group), captive animals offered a diet of dry dog food and apples ad libitum (ad lib group), and captive animals fed restricted amounts of the same diet (maintenance group) were related to growth rates (measured as body mass and head length) of their young. There were no significant differences in the concentration of milk solids among the three groups, but the wild group had higher lipid concentrations and the maintenance group had higher carbohydrate but lower protein concentrations. For all three groups, milk total solids increased through lactation from ca. 25% to ca. 45% and carbohydrate concentrations decreased from ca. 18% to about 3%. Protein concentrations increased from ca. 5% to ca. 10% in the wild and ad lib groups, but only from 4% to 8% in the maintenance group. Lipid concentrations increased in the wild and ad lib groups from ca. 4% to ca. 18%, but in the maintenance group only to ca. 7%. Calculated gross energy content of milk increased through lactation in the wild and ad lib groups (from ca. 500 kJ.100 ml(-1) to ca. 1,000 kJ.100 ml(-1)), but there was no significant increase in the maintenance group. The volume of milk produced increased to a peak just prior to permanent pouch vacation by the young, when the gross energy output in milk was 120-150 kJ.3 h(-1) in the wild and ad lib groups. On a daily basis this is equivalent to the milk energy output of larger wallabies, and helps to explain the relatively high growth rates of young Tasmanian bettongs. There were significant differences in growth rates among the groups, with the heaviest young always in the ad lib group. Thus differences in milk composition resulting from different planes of nutrition can lead to differences in growth rates of marsupial young. PMID- 12624651 TI - Cholinergic innervation to the upper esophageal sphincter muscle in the eel, with special reference to drinking behavior. AB - To elucidate innervation in the upper esophageal sphincter (UES) muscle of the eel, a key muscle in swallowing, repetitive electrical field stimulation (EFS; 30 mA, 40 V, 300 micros, 10 Hz, 10 trains) was employed. Anatomically, the eel UES muscle consists of striated fibers. The EFS-induced contraction of the UES was completely blocked by tetrodotoxin and curare, and abolished in Ca2+ -free Ringer solution. These results suggest that the EFS stimulates nerve fibers specifically and releases acetylcholine as a neurotransmitter. In fact, acetylcholine and carbachol constricted the UES in a concentration-dependent manner. Even after blocking neuronal firing with tetrodotoxin, acetylcholine constricted the UES muscle, suggesting the existence of acetylcholine receptors on the UES muscle cells. Both EFS- and carbachol-evoked contractions of the UES were blocked by curare at a lower concentration than by atropine or hexamethonium, suggesting that the acetylcholine receptor is nicotinic. Even in Ca2+ -free Ringer solution, a direct current stimulus (2 s duration) constricted the UES muscle to an extent similar to that in the presence of Ca2+, indicating that the muscle contraction itself does not need extracellular Ca2+, i.e., the muscle can be constricted by a release of Ca2+ from the sarcoplasmic reticulum. PMID- 12624652 TI - How important is milk for near-weaned red kangaroos ( Macropus rufus) fed different forages? AB - Red kangaroos (Macropus rufus) are large (>20 kg) herbivorous marsupials common to the arid and semi-arid regions of inland Australia, where drought is frequent. Young-at-foot (YAF) red kangaroos are the age/size class usually most affected by drought. Kangaroos at this YAF stage are making the transition from a milk-based diet to one of herbivory and an inability to adequately digest high-fibre feeds may contribute to their high mortalities during drought. We examined the role of milk in the nutrition of YAF red kangaroos fed forages of different fibre content and evaluated it as an extra energy and/or nitrogen source. Milk intake had little impact on the digestion of herbage by YAF red kangaroos fed low-fibre chopped lucerne (alfalfa) hay. Organic matter (OM) intake was 210+/-20 g day(-1) and 228+/-22 g day(-1), respectively, by YAF fed lucerne and lucerne with milk. Apparent digestibility of lucerne OM was ca. 55%, regardless of milk intake. Fed lucerne, with and without milk, YAF sustained growth rates of ca. 45 g day(-1). Conversely, even with a milk supplement, YAF red kangaroos ingested only 90+/-11 g day(-1) of high-fibre chopped oaten hay, of which they digested only ca. 36%. Despite milk intake, YAF fed chopped oaten hay lost between 0 and 75 g body mass day(-1) and were in negative nitrogen balance (-0.40+/-0.11 g N day(-1)). On all diets nitrogen loss was primarily as endogenous nitrogen (urinary and faecal) rather than as dietary nitrogen. Endogenous nitrogen losses were elevated in YAF fed chopped oaten hay, primarily as non-dietary faecal nitrogen. Overall, when high-quality feed was available, YAF were not markedly dependent on milk. However, YAF fed poor-quality chopped oaten hay would require up to 540 ml day( 1) of late-stage kangaroo milk to attain intakes of energy and nitrogen, and hence growth rates, comparable with those YAF fed lucerne. PMID- 12624653 TI - Dietary nutrient levels regulate protein and carbohydrate intake, gluconeogenic/glycolytic flux and blood trehalose level in the insect Manduca sexta L. AB - This study examined the effects of dietary casein and sucrose levels on nutrient intake, and distinguished the effects of carbohydrate and protein consumption on growth, fat content, pyruvate metabolism and blood trehalose level of 5th instar Manduca sexta larvae. Growth increased with increasing casein consumption but was unaffected by carbohydrate intake. Fat content also increased with carbohydrate consumption, but on carbohydrate-free diets fat content increased with increased protein consumption. Blood trehalose level and pyruvate metabolism were examined by nuclear magnetic resonance spectroscopy analysis of blood following administration of (3-(13)C)pyruvate. On diets containing sucrose, blood trehalose increased with increasing carbohydrate intake, and on most diets trehalose was synthesized entirely from dietary sucrose. Pyruvate cycling, indicated by the alanine C2/C3 (13)C enrichment ratio, increased with carbohydrate consumption reflecting increased glycolysis, and pyruvate decarboxylation exceeded carboxylation on all sucrose diets. Larvae that consumed <75 mg/day sucrose were gluconeogenic, based on the [2 (trehalose C6)(Glx C3/C2)]/alanine C2] (13)C enrichment ratio. On carbohydrate-free diets, blood trehalose levels were low and maintained entirely by gluconeogenesis. Blood trehalose level increased with increasing protein intake. Pyruvate cycling was very low, although many insects displayed higher levels of pyruvate decarboxylation than carboxylation. All gluconeogenic larvae displayed alanine (13)C enrichment ratios <0.35 and had blood trehalose levels <50 mM. PMID- 12624654 TI - Na+ gradient-dependent transport of hypoxanthine by calf intestinal brush border membrane vesicles. AB - The properties of hypoxanthine transport were investigated in purified brush border membrane vesicles isolated from calf proximal and distal jejunum. Hypoxanthine uptake in the vesicles was stimulated by a transmembrane Na(+) gradient and an inside negative potential resulting in a transient accumulation of intravesicular hypoxanthine, especially in the proximal jejunum. Na(+) dependent hypoxanthine uptake at this site seemed to occur by two saturable transport systems, a high affinity (K(m)=0.33 micromol/l) and a low affinity (K(m)=165 micromol/l) transporter. Guanine, hypoxanthine, thymine and uracil inhibited intravesicular hypoxanthine uptake, whereas adenine and the nucleosides inosine and thymidine were without effect. These findings represent the first demonstration of active Na(+) gradient-dependent nucleobase transport in intestinal brush border membrane vesicles. PMID- 12624655 TI - Lack of immunological responsiveness to photoperiod in a tropical rodent, Peromyscus aztecus hylocetes. AB - Non-tropical rodents undergo seasonal changes in immune function and disease. It has been hypothesized that seasonal fluctuations in immunity of non-tropical rodents are due to suppressed immune function during harsh winter conditions. A logical extension of this hypothesis is that seasonal changes in immunity should be reduced or absent in tropical rodents that do not experience marked seasonal fluctuations in environmental conditions; however this hypothesis remains to be tested. The present study tested the effects of photoperiod on humoral and cell mediated immune function of male Aztec mice ( Peromyscus aztecus hylocetes). P. a. hylocetes were housed in long (L:D 16:8) or short days (L:D 8:16) for 10 weeks. Animals were then immunized with the antigen keyhole limpet hemocyanin (KLH). Serum anti-KLH immunoglobulin G (IgG) concentrations and splenocyte proliferation in response to the T-cell mitogen Concanavalin A were assessed. Short-day P. a. hylocetes did not display differences in reproductive or immune measures compared with long-day mice. Collectively, these results suggest that P. a. hylocetes are reproductively and immunologically non-responsive to photoperiod. This lack of immunological responsiveness is likely due to the relative seasonal stability of their environment compared with temperate zone species. PMID- 12624656 TI - Oxalate and calcium oxalate mediated free radical toxicity in renal epithelial cells: effect of antioxidants. AB - In a previous study we demonstrated that oxalate induced free radical injury can promote calcium oxalate stone formation. In the present study, we tested whether the antioxidants vitamin E, superoxide dismutase (SOD), catalase and desferoxamine (DFO) can provide protection against oxalate toxicity in LLC-PK(1) cells. LLC-PK(1) cells were exposed to oxalate (1.0 mM) or oxalate+calcium oxalate monohydrate crystals (COM, 500 microg) for 3, 6, and 9 h. Cellular injury was assessed by lactate dehydrogenase (LDH) release. Malondialdehyde (MDA) content, catalase and glutathione peroxidase activities were also measured. The effect of vitamin E (200 microM), DFO (1.0 mM), SOD (400 U), and catalase (400 U) on oxalate-exposed cells was tested. LLC-PK(1) cells exposed to oxalate showed a significant increase in LDH release and MDA content, which was further elevated when COM crystals were added. Cellular glutathione peroxidase and catalase activities were decreased on exposure to oxalate. The addition of vitamin E, SOD, catalase and DFO significantly reduced the release of LDH and restored glutathione peroxidase and catalase activities towards the control level. The increased formation of MDA on oxalate or oxalate+COM toxicity was restored towards normalization by antioxidants and antioxidant enzymes. The protection rendered by vitamin E was greater than that of SOD, catalase and DFO. We conclude that oxalate associated free radical injury may promote stone formation by providing cellular debris for crystal nucleation and aggregation and augment crystal attachment to other tubular cells. Antioxidant administration may prevent calcium oxalate nucleation and retention in the renal tubules by preventing oxalate mediated peroxidative injury. PMID- 12624657 TI - Effects of Tamm-Horsfall protein on the protection of MCDK cells from oxalate induced free radical injury. AB - Renal cell injury and fixed particle formation is one of the theories of urinary stone formation. The exposure of renal epithelial cells to oxalate ions and calcium oxalate monohydrate crystals can cause free radical generation and increase lipid peroxidation. Tamm-Horsfall protein (THP) has a protective effect on the production of free radicals in vitro. We aimed to show that THP (and its deglycosylated products, D-THP) could protect culture cells from free radical injury in vivo as well as the possible mechanism by which this is done. Exposure of Madin-Darby canine kidney (MDCK) cells to Ox resulted in a significant increase in the release LDH, NBT and MDA, as well as an increase in caspase 3 activity, all of which were further elevated when COM crystals were added. With the addition of THP at 500 nM, there was a significant decrease in the release of LDH and the production of MDA and NBT. A decrease in capase 3 activity was observed when 500 nM THP was added to the culture medium that reached 32.7% and 40.4% of inhibition in CaOx+THP and CaOx+COM+THP, respectively. THP decreased the adhesion of COM crystals to the MDCK cells but lost its effect when THP was deglycosylated. The results indicate that both Ox and COM crystals cause the release of LDH, MDA, NBT and increase the activity of capase 3 in MDCK cells. As a free radical scavenger, THP reduces the amount of free radicals and provides significant protection at a critical concentration of 500 nM. The deglycosylated THP decreased the effect of the protection of the MDCK cells from oxalate-induced injury and an increase of adhesion of the COM crystals to the MDCK cells. Therefore, the effects of THP on the protection of oxalate induced radical injury may be partly due to its intact glycosylation and its adhesion to the cell membrane. PMID- 12624658 TI - Targeting gene expression of the mouse uroplakin II promoter to human bladder cells. AB - Differential expression of the desired gene product in the target tissue is central to the concept of gene therapy. One approach is to use a tissue-specific promoter to drive therapeutic genes. To investigate the feasibility of tissue specific gene therapy for bladder cancer using the mouse uroplakin II (UPII) promoter and its transcriptional control, the efficacy of this promoter as well as fragments in regulating gene expression were qualitatively and quantitatively analyzed in bladder and non-bladder tissue cell lines using DNA transfection. Our results demonstrate that the mouse UPII promoter actively drives gene expression in BIU-87, a bladder cancer cell line. Little promoter activity was detected in the non-bladder tissue cell lines. Furthermore, deleting the 5' end 1.5 kb of the UPII promoter by PCR, the activity was significantly decreased but was bladder specific. However, deleting the 3' end 143-bp of the UPII promoter, the activity was hardly detected in any tissue cell lines. The activity of the 3' end 143-bp of the UPII promoter was detected in both bladder cancer and stomach cancer cell lines. These data demonstrate that the mouse UPII promoter has a high activity in human bladder cells and a low basal activity in human non-bladder cells. This suggests that targeting the gene expression of the mouse UPII promoter could be used to treat human bladder cancer. The enhancer was contained in the region of the 1.5 kb of the 5' end of the mouse UPII promoter. The core promoter was located in the region of the 143 bp of the 3' end. PMID- 12624659 TI - Subtotal obstruction of the male reproductive tract. AB - Bilateral obstruction of the male reproductive tract is suspected in men with azoospermia, normal testicular volume and normal FSH. A testicular biopsy is required to differentiate between an obstruction and a testicular insufficiency. Unilateral or subtotal bilateral obstructions and epididymal dysfunction may cause severe oligozoospermia in men with a normal spermatogenesis. However, information on spermatogenesis in oligozoospermic men is lacking, since testicular biopsy is not routinely performed. Men with a sperm concentration of <1 x 10(6) spermatozoa/ml were investigated for possible partial obstruction by performing a testicular biopsy under local anaesthesia. Spermatogenesis was determined by the Johnsen scoring method. A testicular biopsy was performed in 78 men with severe oligozoospermia. The medical history showed male accessory gland infection in 12.8%, previous hernia repair in 14.1% and a history of cryptorchidism in 12.8%. A normal or slightly disturbed spermatogenesis (Johnsen score >8) was present in 39/78 (50%) of the men. Hernia repair occurred more often in men with normal spermatogenesis. A varicocele was predominantly seen in men with a disturbed spermatogenesis. FSH was significantly lower ( P<0.0001) in men with normal spermatogenesis. Subtotal obstruction of the male reproductive tract is a frequent cause of severe oligozoospermia in men with a normal testicular volume and a normal FSH. In other cases, an epididymal dysfunction might explain the oligozoospermia in men with a normal testicular biopsy score. PMID- 12624660 TI - The effect of the somatostatin analogue lanreotide on the prevention of urethral strictures in a rabbit model. AB - We evaluated the effect of the somatostatin analogue lanreotide on the development of surgically induced experimental strictures in the anterior urethra of the male rabbit. A total of 74 male rabbits were randomly allocated into four groups. Lanreotide was administered to the rabbits in groups 2 and 4 from day 0 to 14. To create a stricture, a resection was made in the urethra of the rabbits in groups 3 and 4 on day 2. On day 30, all rabbits were examined with urethrography, impedance planimetry and either histology or for collagen content. Urethrography and impedance planimetry demonstrated a urethral stricture in all operated animals. No difference was found between the two stricture groups, regardless of lanreotide administration, with respect to luminal cross-sectional area (CSA), circumferential tension-strain relation, histology or collagen content. The CSA of the urethra of the normal controls treated with lanreotide was smaller than the CSA of the normal controls not treated with lanreotide, however, no difference was found in histology or collagen content. Lanreotide had no measurable effect on the development of a surgically induced stricture in the male rabbit anterior urethra, however, lanreotide seems to exert an inhibitory effect on the normal growth of the urethra. PMID- 12624661 TI - Differential diagnosis of prostate lesions with the use of biomagnetic measurements and non-linear analysis. AB - The purpose of this study was to investigate the biomagnetic activity emitted from the prostate gland and to differentiate cancerous from benign prostate lesions with the use of biomagnetic measurement and non-linear analysis. Magnetic recordings were obtained from 47 patients with palpable prostate lesions. Histology revealed 24 prostate cancer patients and 23 benign prostate hyperplasia (BPH) patients. The superconducting quantum interference device (SQUID) biomagnetometer was used to measure the prostate's magnetic field by placing the SQUID detector 3 mm above the symphysis pubis. The magnetic field recorded in the 2-7 Hz frequency range was of high amplitude in most malignant lesions whereas all benign cases were of low amplitude. According to our results, the sensitivity, specificity, positive predictive value and negative predictive value were 83.33%, 100%, 100% and 85.18%, respectively. By applying the Grassberger Procaccia algorithm to the magnetoprostatogram time series in malignant and benign prostate lesions we found clear saturation in malignant prostate lesions with m>7 whereas in the benign lesions there was not clear saturation. Prostate cancer emits higher biomagnetic activity than the BPH. This confirms a higher angiogenic activity in prostate cancer than the BPH lesions. Furthermore, the saturation value in the estimation of the correlation dimension of the attractor for the cancer lesions confirms the lower complexity of the system in comparison to the BPH, which is characterized by higher complexity. PMID- 12624662 TI - The X-linked inhibitor of apoptosis protein inhibits taxol-induced apoptosis in LNCaP cells. AB - To clarify the roles of the X-linked inhibitor of apoptosis protein (XIAP), we investigated the effects of XIAP overexpression on taxol-induced cell growth arrest and apoptosis in the prostate cancer cell line (LNCaP). After the transfection of XIAP cDNA into LNCaP cells, we established clonal cell lines that overexpressed XIAP and examined the taxol effects on growth and apoptosis by 2-(4 lodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt and flow cytometric analysis. The effects of XIAP overexpression on caspase 3 were examined by immunoblot analysis and activity assay. The interaction between XIAP and caspase-3 in LNCaP cells was examined by cotransfection with myc XIAP and caspase-3-HA cDNAs followed by immunoprecipitation and immunoblot analysis. Large amounts of XIAP were expressed in the established cell lines. Although the growth rates were reduced in a dose- and time-dependent manner by taxol, these effects were significantly decreased in XIAP stably overexpressing cell lines. In addition, we found that taxol treatment induced the cleavage of pro-caspase-3, followed by apoptosis, and that the overexpression of XIAP inhibited apoptosis by attenuating the cleavage of pro-caspase-3 and caspase-3 activity. Interestingly, XIAP bound to pro-caspase-3 in LNCaP cells transiently cotransfected with myc-XIAP and caspase-3-HA cDNAs, and this interaction was inhibited by taxol treatment. These results suggest that the overexpression of XIAP inhibits taxol-induced apoptosis through the decrease of caspase-3 activity and inhibition of the processing of pro-caspase-3. PMID- 12624663 TI - Changes in the practice of non-invasive ventilation in treating COPD patients over 8 years. AB - OBJECTIVE: We reviewed data of 208 episodes of acute respiratory failure due to chronic obstructive pulmonary disease treated by non-invasive ventilation (NIV) in our Respiratory Intensive Care Unit (RICU) from its opening in 1992 to 1999. MATERIAL AND METHODS: We assessed whether the rate of NIV success, the severity of the disease, and the associated costs changed in this period during which the staff and the equipment did not change. RESULTS: The failure rate was constant over the years (17.2% on average). The severity of the episodes of ARF, defined by pH and APACHE II at admission, worsened during the years. The statistical change point test allowed us to identify 1997 as the year of a significant change in the severity of admission pH and therefore to identify two different periods: 1992-1996 (mean pH = 7.25+/-0.07) and 1997-1999 (7.20+/-0.08; P<0.001). In this latter period the risk of failure for a patient with a pH <7.25 was threefold lower than in 1992-1996. In 1997-1999 an increasing number of episodes of ARF with a pH >7.28 were treated in the Medical Ward (20% vs 60%). This allowed a significant reduction of daily cost per patient treated with NIV (558+/-8 Euros vs 470+/-14 Euros, respectively; P<0.01). CONCLUSIONS: We conclude that, over time, experience with NIV may progressively allow more severely ill patients to be treated without changing the rate of success. The daily cost of NIV per patient can be reduced by treating less severely ill patients outside the RICU. PMID- 12624664 TI - Hypothermia protects against endotoxin-induced acute lung injury in rats. AB - OBJECTIVE: Hypothermia in humans and animals is known to decrease the number and function of circulating neutrophils. Because an activation of circulating neutrophils and their sequestration into the lung are important pathogenetic phenomena in endotoxin-associated lung injury, we conjectured that hypothermia could prevent this type of lung injury. DESIGN AND SETTING: Animal study at a university-affiliated research institute. SUBJECTS: Thirty-six Sprague-Dawley rats. INTERVENTIONS: After anesthesia, the rats were randomly assigned to normothermia (37 degrees C, rectal temperature) or hypothermia (27 degrees C), which was induced by surface cooling. After 1 h of stable temperature, the rats were administered intratracheal doses of lipopolysaccharide (LPS; 3 mg/kg) (normothermia-LPS; hypothermia-LPS) or an equivalent volume of normal saline (normothermia-saline; hypothermia-saline). The rectal temperature was maintained within +/-1 degrees C of the target temperature for 6 h after the intratracheal treatment. MEASUREMENTS AND RESULTS: Compared with the normothermia-LPS group, the neutrophil count in bronchoalveolar lavage (BAL) fluid (p=0.002) and the myeloperoxidase activity of lung tissues (p=0.002) of the hypothermia-LPS group were both lower. Compared with the normothermia-LPS group, the BAL interleukin 1beta level of the hypothermia-LPS group was lower (p<0.001), whereas the BAL interleukin-10 level of the hypothermia-LPS group was higher (p=0.026). Compared with the normothermia-LPS group, the histologic scores for acute lung injury of the hypothermia-LPS group were lower (p=0.007). CONCLUSIONS: Hypothermia pretreatment decreased the pulmonary sequestration of neutrophils, induced a favorable balance between pro- and anti-inflammatory cytokines, and attenuated histologic injury in endotoxin-challenged rats. PMID- 12624665 TI - Quality control of mechanical ventilation at the patient's home. AB - OBJECTIVE: During home mechanical ventilation the prescribed settings are applied without permanent supervision of health professionals. After a long-time period of unattended operation at home the ventilator may not apply the ventilation parameters prescribed. This quality control study of home mechanical ventilation assessed whether tidal volume (V(T)), frequency (f), and minute ventilation (V'(E)) actually applied by the ventilator coincide with the values set on the ventilator control panel and with those prescribed. MEASUREMENTS: Actual V(T), f, and V'(E) applied by the ventilator in 30 patients on nocturnal HMV were measured at the patients' homes. The patients were subjected to volume targeted assist ventilation through nasal mask (n=28) or tracheostomy (n=2). The values of V(T), f, and V'(E) set at the ventilator were recorded. The actual and set V(T), f, and V'(E) values were compared with those prescribed. RESULTS: Considerable differences were found between actual, set and prescribed V(T), f, and V'(E). Actual V'(E) was significantly lower than V'(E) set: mean difference was 0.82 l/min, with considerable individual differences. Differences between actual and prescribed V'(E) were caused both by a poor performance of the ventilator and by a discrepancy between the values prescribed and those set at the ventilator control panel. CONCLUSIONS: Regularly assessing the actual performance of ventilators at the patient's home is a quality control procedure useful for detecting malfunctions which could improve compliance and outcome of home mechanical ventilation. PMID- 12624666 TI - [Epidemiology of colorectal cancer]. AB - Colorectal tumors are among the most frequently encountered forms of cancer worldwide. With approximately 57,000 new cases every year, they represent the most frequent type of cancer in Germany, ranking before breast cancer (approximately 46,000) and lung cancer (approximately 37,000). Although global incidence is on the rise, in Germany it is only increasing among men, but not among women. The mortality rate (approximately 26,500 deaths annually) in Germany has declined among men for about the past 10 years and among women for about the past 20 years. The most important risk factors are familial history of colorectal and other tumors as well as lifestyle factors such as nutrition, obesity, inactivity,and smoking.Lifestyle-related risks offer a broad area for implementing primary preventive measures,which have not yet been adequately exhausted. Several proven (fecal occult blood test) and probably effective (endoscopic) methods are available for secondary prevention. Consistent encouragement of these possibilities for prevention could reduce incidence and mortality substantially and render colorectal tumors less frequent. PMID- 12624667 TI - [Diagnosis of colorectal carcinoma. An update]. AB - Colorectal cancer (CRC) is one of the most frequent tumors in western countries. More than 50% of all CRC are diagnosed at an advanced stage which precludes curative treatment. For this reason, early detection of CRC is mandatory to improve longterm outcome. Fecal occult blood testing (FOBT) once per year and subsequent colonoscopy (if the FOBT is positive) provides up to 30% decrease in mortality from CRC. Due to the fact that current data indicate a 60% reduction in CRC-associated mortality, colonoscopy has recently been approved for CRC screening by german public health insurance companies. Yet efforts in screening largely depend on patient compliance, particularly in view of cost-effectiveness. Introduction of new imaging techniques (CT-/MRI-colonography) may increase general acceptance, but clinical benefit and costs still remain to be determined in larger studies. PMID- 12624668 TI - [Importance of coloncontrast enema]. AB - Screening for colorectal cancer starts with digital exploration of the rectum and was extended in 1977 using the haemoccult test. It was referred to detect 24% of malignancies, but with a high number of false positive findings. Combination of regular follow up examinations and coloscopy reached a sensitivity of 80%. International studies demonstrated the need of a complete colon examination, so that the coloscopy is actually an important screening method in Germany. The coloncontrast examination is a safe, cost-effective method with a low complication rate. It is suitable especially for patients, in which the colonoscopic evaluation of the colon was not complete and provides additional morphological informations in many cases. PMID- 12624669 TI - [Importance of CT and MRI in the follow-up of patients with rectal cancer]. AB - INDICATION: Presentation and evaluation of diagnostic procedures for the follow up in patients with suspected local recurrence of rectal cancer. METHODS: Follow up usually is performed using endoscopy and endosonography. Imaging techniques applied are also contrast enema as well as helical CT,MR imaging (MRI), and positron emission tomography (PET). RESULTS: The results demonstrate the difficulty of all available modalities to differentiate between early tumor recurrences from post-therapeutic alterations. Especially the differentiation of scar tissue and local tumor recurrence, the detection of lymph node as well as pulmonary or hepatic metastasis requires a large diagnostic effort with employment of various imaging modalities. Values determined for sensitivity and specificity for CT are 84% and 91% with a positive predictive value (PPV) of 78%. For MRI and PET the sensitivity are 94% and 98%,and the specificity 92% and 90% with a PPV of 93% for the PET. CONCLUSION: The results demonstrate the increasing importance of CT and MRI in the follow-up of patients with rectal cancer and enable an accurate and early diagnosis through employment of new examination strategies and sequence protocols.PET remains an add-on examination in uncertain local lesions. PMID- 12624670 TI - [Diagnosis of colorectal tumors]. AB - With the introduction of multislice CT extensive volumetric data sets can be quickly acquired in high spatial resolution. The high spatial resolution reduces partial volume effects and enables multiplanar reconstructions. Regarding the colorectum this means that the colon can be assessed if the colon is sufficiently cleaned and distended, and that transmural infiltration of colorectal carcinoma and liver metastases can be better detected. T-staging of colon cancer is less important than T-staging of rectal cancer. Based on the higher contrast MRI is superior to CT in T-staging of rectal cancer and in the differentiation between scarring tissue and recurrence of carcinoma. PMID- 12624671 TI - [Early detection of colorectal tumors: CT or MRI?]. AB - Cancer screening is currently offered for the organs breast, prostate, cervix and colorectum. With progressing technique in computerized tomography (CT) and magnetic resonance imaging (MRI) the colorectum can be increasingly better assessed. Because in CT and MRI also other organs are imaged with the colorectum, a colorectal screening automatically becomes a multiorgan screening. CT and MRI protocols designed for early detection of colorectal tumors are presented and discussed with regard to monoorganscreening (in CT: low dose, no i.v. contrast) and multiorganscreening (in CT: diagnostic dose for neighboured organs with i.v. contrast). More information under http://www.multiorganscreening.org. PMID- 12624672 TI - [Predictive parameters of infectiologic complications in patients after TIPSS]. AB - AIM: To define predictive parameters of a complicated clinical course after the TIPSS procedure. METHODS: Blood cultures were drawn prospectively in 41 patients from a central line and from the portal venous blood before stent placement as well as from the central line 20 min after intervention. C-reactive protein (CRP) (mg/dl) and white blood cell count (WBC,/microl) on the day of TIPSS-procedure (d0), the first (d1) and seven (d7) days after TIPSS were compared in patients with a complicated clinical course (spontaneous bacterial peritonitis,pneumonia,sepsis; group I) to patients without clinical complications (group II) RESULTS: Group I showed a significant increase in CRP (d0: 1.8+/-1.0; d1: 3.2+/-1.5; d7: 4.3+/-3.2), and white blood cell count (d0: 7700+/-2600; d1: 10800+/-2800; d7: 7500+/-1800) on the first day after TIPSS-procedure in comparison to group II (CRP: d0: 1.6+/-0.6; d1: 1.8+/-1.0; d7: 1.9+/-0.6. WBC: d0: 6900+/-1500; d1: 8000+/-1600; d7: 7600+/-1400). Microbiological analysis showed in 12% skin or oral flora in the last sample. CONCLUSIONS: The course of CRP and WBC-count during the first week after TIPSS procedure may indicate patients with a potential risk of a complicated clinical course. PMID- 12624673 TI - [Diagnostic imaging and differential diagnosis of pathological processes of the sinus cavernosus]. AB - To describe the different imaging modalities for the evaluation of pathological changes in the cavernous sinus as well as to compile criteria for differential diagnosis. Imaging of the cavernous sinus comprises the primary use of tomographic modalities such as CT or MRI. The continuing development of multislice-CT (MSCT) allows the depiction of bony structures of the cavernous sinus with a high resolution.Secondary reconstructions of the acquired data set allow a reliable evaluation especially of the bony topography including the foraminae of nerves and vessels. Uni- or bilateral structures can be visualized using contrast-enhanced CT.CT-angiography is capable of demonstrating the course of the internal carotid artery and its involvement in pathologies of the cavernous sinus, this recent achievement has only been made available by use of the current short scanning times. Contrast-enhanced MRI in axial and coronal orientation, optionally using fat saturation techniques as well as arterial and venous MRI-angiography (MRA) are used to depict the soft parts of the cavernous sinus. Care must be taken to cover all topographic detail including different signal intensities, as multiple inborn, neoplastic, infectious or traumatic changes can be present hampering the radiologic diagnosis of the cavernous sinus. Uni- and bilateral infiltration of the cavernous sinus as well as vascular involvement represent additional criteria leading to the differential diagnosis. The advent of dynamic sequences as well as diffusion and perfusion weighted MRI have broadened the spectrum of diagnostic modalities. Digital subtraction angiography as an invasive technique is used during therapeutic procedures such as the local treatment of aneurysms or carotid-cavernous fistulas. PMID- 12624674 TI - [Multiphasic MR angiography as an intra-individual comparison between the contrast agents Gd-DTPA, Gd-BOPTA, and Gd-BT-DO3A]. AB - QUESTION: The availability of new MR contrast agents having either a protein binding effect or higher concentration leads to the question if they differ from standard compounds in their vascular contrasting properties. METHODS: By intraindividual comparison five volunteers were examined (1.5 T MR system) with a multi-phasic MRA of the abdomen each receiving three different contrast agents (gadopentetate dimeglumine 0.5 M; gadobenate dimeglumine 0.5 M, and gadobutrol 1.0 M). The dose (0.15 mmol/kg body weight) and flow rate (3.0 ml/s) were kept identical. All images were quantitatively and qualitatively evaluated by blinded assessment. RESULTS: Significantly higher maximum signal intensity was found in the arteries for the protein interacting gadobenate dimeglumine (p = 0.05). No significant difference in vascular enhancement was demonstrated in the comparison of gadobutrol to gadopentetate dimeglumine. CONCLUSION: Gadobenate dimeglumine was shown to be a favorable contrast agent for multi-phasic MRA. A higher concentrated Gd-chelate does not automatically lead to improved vascular contrast if standard imaging protocols are used. PMID- 12624675 TI - [Diagnosis of thyroid disease. Part 2: thyroid gland nodes, diagnosis and treatment of thyroid carcinoma]. PMID- 12624677 TI - [Current preclinical findings on substances against Parkinson's disease]. AB - Striatal dopamine loss provides the premise for dopamine substitution in palliative therapy for Parkinson's disease (PD). This includes firstly L-dopa (L 3,4-dihydroxyphenylalanine, levodopa) and dopamine receptor agonists. Although this therapy has been demonstrated to induce marked clinical improvement in the early stages of PD, its use is limited in the long term by a loss of efficacy. In addition, most patients develop the "long-term L-dopa syndrome," which is characterized by a decrease in the control of Parkinsonian symptoms (decrease in the drug's effect) and the appearance of certain motor disturbances including episodes of akinetic freezing, debilitating dyskinesias, and on-off periods. The aim in developing new drugs is to achieve better therapeutic approaches. In the case of PD, the main strategies are to develop (1) alternatives to L-dopa therapy for alleviating the striatal dopamine deficit while avoiding or retarding the long-term L-dopa syndrome, (2) antidyskinetic approaches, and (3) neuroprotective drugs aimed at causal treatment of PD which is able to preserve the remaining dopaminergic neurones and to halt or at least retard the disease process. This article reviews new approaches for the treatment of PD and presents findings from our studies using a new experimental in vivo model of PD. PMID- 12624678 TI - [Methods for the early diagnosis of Parkinson's disease]. AB - Patients with idiopathic Parkinson's disease display a 60% degeneration of the nigrostriatal neurons before motor symptoms have progressed enough to allow clinical diagnosis. It is clear that any neuroprotective therapy starting at such a late stage can have no substantial effect on the disease progression. Therefore, earlier diagnosis must be the goal of future research,when at most mild motor or non-motor symptoms are present or when only risk factors can be identified. Evidence of various gene mutations associated with idiopathic Parkinsonism raise the hope that these or other biological markers will allow earlier identification of patients at risk. A possibly significant vulnerability factor for developing Parkinson's disease can also be demonstrated by means of transcranial sonography. Since the individual tests are not sufficiently specific or sensitive, a gradual, precise, and inexpensive battery of tests needs to be developed for the successful identification of a risk group for this disease. The extent of damage to the dopaminergic system in these patients can be quantified using nuclear techniques. PMID- 12624679 TI - [Medicinal treatment of idiopathic Parkinson's disease]. AB - Idiopathic Parkinson's disease (IPD) results from a largely selective degeneration of nigrostriatal dopaminergic neurons. Therefore, compounds which strengthen dopaminergic transmission in the striatum are the most important therapeutic approach. These include L-dopa, dopamine receptor agonists, selegeline, and entacapon. Since nigrostriatal degeneration leads to secondary alterations of cholinergic and glutamatergic transmission to the basal ganglia,nondopaminergic compounds such as anticholinergics and N-methyl-D aspartate (NMDA) receptor antagonists are also used in the management of IPD. L dopa is the most effective substance but after 3-5 years of L-dopa treatment, approximately half of all IPD patients develop fluctuations. Therefore, initial treatment with a dopamine receptor agonist is recommended. PMID- 12624680 TI - [New approaches to transplantation of dopamine-producing cells and stem cells]. PMID- 12624681 TI - [Management of fractures in the severely injured--influence of the principle of "damage control orthopaedic surgery"]. AB - There have been substantial changes in the management of multiply injured patients over the last decades. In the 1950s and 60s, perioperative care was limited and the surgical techniques were not well developed. It was therefore discussed that a patient might be "too sick to operate" and the general recommendation was to postpone surgical care of the extremities,until a patient had stabilized. In addition this recommendation was maintained by fears of the "fat embolism syndrome", which was considered to be directly related to fat and intramedullary contents released from the fracture site leading many to believe that early manipulation of the fracture was unsafe. The use of simple splintage clearly demonstrated the importance of skeletal stabilisation by reducing the effect of any continuing injury and this positive effect of skeletal stabilistion became more obvious with the implementation of standardized techniques of osteosynthesis. In the 1970's, pioneer studies appeared in the literature reporting that immediate stabilisation of femur fractures drastically reduced problems of traumatic pulmonary failure and post-operative care when compared to traditional non-operative fracture management. Thereafter, immediate and complete definitive operative care of all fractures has been said to represent the optimal treatment for the patient with multiple orthopaedic injuries and the benefits of this approach have been demonstrated in numerous studies within the last two decades.However,exceptions have been discussed in the past few years, where the principle of early total care may not be beneficial (head and chest trauma, high ISS predisposing to posttraumatic complications, borderline patients). In high risk, borderline patients, the surgical burden was demonstrated to increase the risk of postoperative complications. For these patients, the concept of initial temporary fixation and secondary conversion to a definitive procedure has recently been advocated, and the term "damage control orthopaedic surgery" was coined. Within recent years, an increased consensus has occurred across the countries and the oceans in regards to the acceptance of the concept of damage control. This manuscript documents the pathogenetic back grounds and the clinical results leading to a change in the management of orthopaedic injuries towards damage control. PMID- 12624682 TI - [Tibial plateau fracture--biodegradable bonecement-augmentation]. AB - Between October 1996 and January 1999,29 patients (f:16,m:13,age: 22-86) with fractures of the lateral tibial plateau were operated on arthroscopic,fluoroscopic control or were treated with open reduction and internal fixation. 15 of them were retrospective and 14 prospective analysed. The metaphyseal defect after elevation of the depressed fragment was augmented in 11 cases with autologous spongeous bone grafting,in 9 cases with biodegradable bone cement (Norian SRS). Augmentation was unnecessary in 9 cases. The results according to the Lysholm score and the radiological results were good or excellent. Concerning the kind of augmentation no difference was noted. In the Norian SRS-group the duration of postoperative treatment was shorter than in the other group. The duration of partial weight bearing was shorter too. The results of the present study suggest that an injectable calcium phosphate cement may be a competent material for augmentation in lateral tibial plateau fractures because of the application form and the initial high mechanical stability. PMID- 12624683 TI - [Results of elastic stable intramedullary nailing (ESIN) in forearm fractures in childhood]. AB - Conservative treatment of unstable forearm fractures in childhood leads to disappointing results in up to 50%. Therefore the indication for a primary operative treatment is given. In the early 1990s the ORIF (open reduction and internal fixation) Procedure was the commonly practiced treatment. Nowadays elastic stable intramedullary nailing (ESIN) is increasingly recommended. Aim of our prospective investigation was to evaluate difficulties, early results and complications of the elastic stable intramedullary nailing of forearm fractures in children. From January 1998 to December 2000 fifty-four children (23 female, 31 male) with an average age of 8 years (8-16) were treated operatively with ESIN. 48 patients had a single forearm-fracture,6 were multiple injured (ISS<16). Open reduction was necessary in 7,4% cases. Clinically and radiologically a good consolidation was found in all fractures. In one case a large callus formation induced a reduced range of motion. As complications seven superficial infections,which required neither pharmacological nor surgical intervention,were treated. In two cases a temporary paraesthesia of the N. radialis superficialis occurred. In one patient a progressive lesion of the N. ulnaris was observed, due to a constriction of scar-tissue. All fractures healed, in all cases with full range of motion. No relevant difference of length or deviation of the axis was observed. Good clinical results, the low risk of complications and the positive biological impact make the ESIN to the standard procedure in the treatment of unstable forearm fractures in childhood. PMID- 12624684 TI - [Intraarticular pathology of atraumatic shoulder dislocations. An arthroscopic study]. AB - AIM OF THE STUDY: According to the literature, atraumatic shoulder dislocations occur without or only minor trauma,self-reduction and no or little pain. Little is known about intraarticular pathology in this entity. It was the purpose of our study to evaluate such findings. METHODS: During a 2-year period,226 patients had surgery for shoulder dislocation.28 patients met the criteria for atraumatic dislocation as above (group A). Patients with bony pathology or recurrent microtrauma were excluded. All had been treated with a rehabilitation program without success.28 consecutive patients with surgery after posttraumatic dislocation served as a control group (group B). At the beginning of the surgery,arthroscopy was performed in all patients and the intraarticular findings were recorded. For the capsulo-labral pathology,we determined 3 types: type I had capsular elongation or scarring and included so called "non-Bankart-lesions"; type II had classic "Bankart"-lesions and type III more complex capsulo-labral lesions like e.g. "ALSPA"-lesions. RESULTS: The mean age in group A was 27.6 y and 26.2 y in group B.12 patients in group A and 5 in group B were female. The average no.of dislocations was 10 (1-30) or 9 (1-28), respectively. In group A we saw type I lesions in 11 patients (39.3%), type II lesions in 9 (32.1%) and type III lesions in 8 (28.6%) patients. In group B we found type I lesions in 8 (28.6%), type II lesions in 4 (14.3%) and type III lesions in 16 (57.1%) patients.Hill-Sachs lesions were found in 22 (78.6%) and 23 (82.1%) of the patients, respectively.Also, we saw chondral glenoid damage, cuff lesions and SLAP-lesions in both groups. CONCLUSION: The above mentioned criteria for atraumatic shoulder dislocation do not exclude intraarticular pathology comparable to posttraumatic cases, at least for patients, who do not respond to a conservative treatment. Besides the etiology, the given pathology must be considered for therapy. PMID- 12624685 TI - [Refixation of the meniscus with the SD meniscal staple: a comparative biomechanical study on cadaver knees]. AB - GOAL: The failure load of the SD meniscal staple was compared with the 2-0 Etibond meniscus suture in a dynamic test without isolating the meniscus. METHODS: In eight knee joint pairs, a standardized lesion of the posterior part of the medial meniscus was repaired by either three staples (7 mm) or three 2-0 Etibond sutures. After resection of the cruciate ligaments and 15 degrees external rotation, 45 degrees flexion, and 70 kg of axial loading, a tibial translation was exerted on the knee joint. The test was documented on a way force diagram, demonstrating the failure of the device at a sudden load of force. RESULTS: The average load before failure of the meniscus staple was 591.2 N (540 690 N), that of the sutures 850 N (600-1350 N). We did not find any statistical correlation to age,weight, or height of the specimens. The suture migrated, but never ruptured. In two cases the suture caused a radial meniscus lesion. Failure of the staple was detachment of the inferior part in all cases, but no migration. CONCLUSION: This test is based on the dynamic forces of the knee joint on the meniscus. Our measurements show lower failure loads of the staple compared to those of the sutures, but compared to other tests a stable method of meniscus repair. PMID- 12624686 TI - [Comparative examination between an angle stable, monokortikal osteosynthesis technique with the conventional plate osteosynthesis at the anterior arm shaft fracture]. AB - With a retrospective Follow-up essay under the use of Matched peer variables the wellbeing course was examined by patient pairs (128 patients) assigned to 64 individually with fractures of the anterior arm shaft. A group which treats others with 2.7 or 3.5 mm DCP/LC-DCP got with the AO Point Contact basic gate (PC Fix). The well-being course data count after the implantation on a period of 18 months. The Follow-up-rate was 100% for this time period. The patients became for each other on reason of the criteria: assigned to fracture classification, soft partial loss, accompanying injuries and age. Possible complications were: implantation conditional nerve damages, infections, implant failures, delayed healings, pseudarthroses, motion reductions and synostoses. Complications appeared (PC-Fix at 13 patients: respect, DCP: 5). The statistical testing didn't yield any statistically significant advantage for one for the two implants at a p value of 0.5811 for. PMID- 12624687 TI - [Clinical impact of MRI in acute wrist injuries. Indication and therapeutic consequences]. AB - PURPOSE: This prospective study was aimed at evaluating the clinical impact of magnetic resonance imaging (MRI) vs conventional radiographs in the early diagnosis of acute wrist fractures. The influence of MRI on the period of being unable to work was demonstrated. MATERIALS AND METHODS: MRI was performed within a mean of 6.6 days after initial radiographs in 54 patients (56 wrists) with clinical suspicion of wrist fractures and normal plain or indistinct radiographs. MRI findings were read without knowledge of the initial radiographs. RESULTS: In 31 of 56 wrists MRI findings resulted in a change of diagnosis. There was a false positive diagnosis on plain radiographs in nearly one-half ( n=25) of the cases,whereas only six cases had false negative results. In 28 cases MRI allowed the detection of additional injuries of soft tissues. In 35 of 56 cases radiological follow-up was no longer necessary. In 22 of 54 patients the period of immobilization could be shortened or treatment discontinued ( n=18). The period of being unable to work was shortened in 16 of 54 patients, while in 7 of 54 patients this time span had to be prolonged for therapeutic reasons. In 31 of 54 patients MRI-based therapeutic consequences had no influence on the period of being unable to work. CONCLUSION: Our results show a high clinical impact of MRI in the detection of acute wrist fractures. Early MRI is able to reduce economic costs due to efficient therapeutic treatment and shortened periods of being unable to work. We recommend MRI immediately on the day of trauma in the presence of clinical suspicion and equivocal plain radiographs PMID- 12624688 TI - [Results on treatment of anterior cruciate ligament rupture of immature and adolescents]. AB - Between 01.01.1994 and 31.12.2000 we have operated 45 patients in our clinic with open growth plates. Cruciate ligament injuries are seldom found by children with open growth plates.Nevertheless, if the injury is left untreated it is liable to injure the meniscoid and the cartilage severely and may even lead to a loosening of the tertiary stabilizers. Therefore we do refixation of anterior cruciate ligament for tibial bony rupture and Refixation in Marschall technique or a replacement for the anterior cruciate ligament using Hamstring ligaments and fixated them by using the transfix technique.Up till now, none of the patients showed any signs of ill-functioning growth plates. Though it may be necessary to point out that the patients had not fully grown out by the time the subsequent check ups were held. The knees we revised were all stable and after relating our results to the IKDC-Score we found that our treatment reflected a good to very good result (Group A and B). Results of Marschall-Technique are very poor, results after bony refixations are good, result of our prospective study encourage us to recommend the surrogate of the anterior cruciate ligament by using the transfix technique in cases where the growth plates are still open. Therefore we can recommend reconstruction on transfix technique and refixation of bony ruptures. PMID- 12624689 TI - [Simultaneous reimplantation of both lower legs--5-year follow-up (case report)]. AB - We are reporting the case of a 29 year old male in whom we performed successful reimplantaton of both lower legs following trauma inflicted by a railroad boxcar. Five years after this accident, the patient's walk is almost normal and both deep sensitivity and two point discrimination on the soles of his feet are sufficient. The patient can walk, run and stand very well on one leg, both on even and on uneven ground.He returned to his job with the railroad 8 months after his accident. Originally the patient was employed as a railroad workman, and is now an office employee. His private life is normal and he enjoys hiking and dancing. In our opinion, sufficient function of the tibial nerve in the reconstructed extremity is important for clinically satisfactory long-term results. Both the Mangled Extremity Severity Score (MESS) and the NISSSA are helpful in making the decision on whether to primarily amputate or reconstruct Gustillo IIIC cases. Good long-term results as well as general cost reduction are achievable following reconstruction of extremities. Amputation of an extremity can be predicted with 100% certainty when MESS is 9 or more. Primary shortening and secondary lengthening of an extremity is a good method of treating Gustillo III C fractures. PMID- 12624690 TI - [Total hip arthroplasty for metastatic Merkel cell carcinoma]. AB - Merkel cell carcinoma is an extremely rare malignant tumor which derives from the neuroendocrine cell system with features of epithelial differentiation. It belongs to the APUD-system (amine and precursor uptake and decarboxylation) and is characterized by highly aggressive spread with a predisposition for local recurrence and local regional and distant metastases. Metastatic spread to the hip has not been described in the literature before. We report on a 76-year old male patient with metastases of the left femoral head and greater trochanter 3 1/2 years after excision of a Merkel cell carcinoma of the left thigh with consecutive radical lymph node dissection of the left inguinal area and radiation therapy. Although microscopic,immunohistological and ultrastructural characteristics of the carcinoma have been well defined, there are no established treatment guidelines and prognostic factors that may predict the behaviour of the tumor due to the limited number of cases. Elective lymph node dissection decreases the rate of local recurrence but is not associated with improved overall survival. At the time of establishing the diagnosis, about half of the patients has positive lymph nodes with a 3-year-survival rate of 60%. PMID- 12624691 TI - [Reconstruction of bone defect by vascularized fibular transfer after failed wrist arthroplasty]. AB - A bone defect of 14 cm resulted after wrist implant removal because of loosening in a 46-year-old patient. Performing wrist arthrodesis the defect was reconstructed by a vascularized fibular transfer. Solid bone fusion was complete in six weeks. Free function of the digits was seen because of length reconstruction. PMID- 12624692 TI - [Penetrating knife injury of the heart. Emergency management and definitive treatment: case report and review of the literature]. AB - The outcome of patients with penetrating heart injuries depends to a great extent on aggressive primary care and fast transport to the closest appropriate trauma center. There, after confirming the diagnosis, the injured victim has to be transferred without any delay to the operating room where the penetrating injury can be dealt with. The importance of resolute emergency management is shown based on a case presentation and a review of the current literature. The employment of extracorporal circulation (heart-lung machine) is rather subordinate in the primary care of these patients. PMID- 12624693 TI - [Serious complication after subcutaneous injection of heparin for prophylaxis of thromboembolism. Case report]. AB - Subcutaneous application of low-dose heparin before and after surgery is a routine procedure to avoid thromboembolic complications. Advances in development of anticoagulants, reduction of intervals between applications, modern injection tools,and training of the personnel have already reduced the incidence of severe complications. However, the case presented shows impressively that a life threatening complication is still possible and has to be kept in mind in perioperative thrombosis prophylaxis. After postoperative subcutaneous injection of low-dose-heparin in the right lower abdominal wall, a 76-year-old female patient suffered from an extensive,hemodynamically active hematoma located in the M.rectus abdominis as a sequela of perforating the A. epigastrica superficialis. Consequently hypovolemic shock led to cardiopulmonary circulatory arrest. After immediate resuscitation, surgical hemostasis was performed and the hematoma was removed. Post-interventional stabilization of circulation and wound healing were trouble free. PMID- 12624694 TI - [Opioid therapy in chronic non-malignant pain]. AB - In long-term treatment opioids seem to have only minimal side-effects compared with other analgesics and co-analgesics.Nevertheless, some risks have to be considered. While immunosuppression, neurotoxicity, teratogenity, tolerance and addiction are clinically not relevant or very rare, cognitive impairment, sedation and obstipation may have a clinical impact.However, these symptoms can usually be managed by adjuvant medication and patient education. Treatment of non malignant pain with opioids can only be considered on an individual basis. Scientific evidence for general treatment with opioids, treatment of specific pain syndromes or treatment with certain opioids is not available. In conclusion, only recommendations regarding opioid treatment for certain chronic pain syndromes can be made. In only a minority of patients can a long-term analgesic effect be expected.Therefore, careful evaluation of alternative options of pain management is necessary before opioid therapy is started. With standardized documentation responders may be distinguished from non-responders. For clinical practice of long-term opioid therapy in non-malignant pain a specialized knowledge in pain management is a prerequisite. Future studies with more sophisticated methodology will be necessary to advocate more precise guidelines.However, the therapeutic recommendations from the DGSS consensus conference allow a safer,well structured and validated use of opioids for chronic non-malignant pain. PMID- 12624695 TI - [Ultrasound-guided central venous cannulation in neuropaediatric patients to avoid measures causing potential increase in brain pressure]. AB - BACKGROUND: Certain measures such as the Trendelenburg position or an increase in intrathoracic pressure raise the chances for a successful puncture of the internal jugular vein (IJV) particularly in paediatric patients. However, these measures are contraindicated in patients with increased intracranial pressure. Therefore, in anaesthetised and ventilated neuropaediatric patients we investigated whether ultrasound-guided cannulation of the IJV can replace these measures. METHODS: In this prospective study,8 newborns, 46 infants and 66 small children were included. The cross-sectional area of the right IJV was sonographically measured in horizontal and 15 degrees Trendelenburg positions with or without PEEP 0, 5 or 10 cm H(2)O. The IJV was then punctured at the conditions of a predetermined position and PEEP combination under sonographic guidance (5-8 MHz). RESULTS: The mean cross-sectional area of the IJV was 0.3-1.6 cm(2) in the horizontal position without PEEP ( n=120) and was enlarged due to the Trendelenburg position by 24% and by 63%, when PEEP 10 cm H(2)O was added. Independent of positioning and PEEP the ultrasound-guided puncture was successful at the first attempt in 98% of cases and a second attempt had to be made in only two children. CONCLUSIONS: Under sonographic visualisation of the cross-sectional area, the IJV can easily and safely be punctured for central venous cannulation in newborn, infants and small children without measures such as the Trendelenburg position or implementation of PEEP. PMID- 12624696 TI - [Prevention of post-operative nausea and vomiting. Randomised comparison of dolasetron versus dolasetron plus dexamethasone]. AB - BACKGROUND: Postoperative nausea and vomiting (PONV) are frequent complications after operations. The aim of this study was to assess the efficacy of combined dolasetron plus dexamethasone versus dolasetron alone with respect to the incidence and severity of emetic symptoms and patients satisfaction. METHODS: In a prospective, randomised, double-blind study, 150 patients scheduled for hysterectomy or breast surgery were allocated to one of the following two groups: group A received 50 mg dolasetron orally and group B 50 mg dolasetron orally plus 8 mg dexamethasone intravenously. The follow-up was for 24 h after surgery. RESULTS: In group A PONV was significantly more frequent (28%) compared to group B (12.0%). The incidence of vomiting was significantly lower in patients receiving dolasetron plus dexamethasone (0%) in comparison to patients receiving dolasetron (8.0%). Furthermore,patients satisfaction was significantly higher in group B compared to group A. About 6 or 7 patients need to be treated with additional dexamethasone instead of a placebo for one patient to benefit from this intervention (i.e. to stay free from PONV) who otherwise would have suffered from PONV. CONCLUSIONS: Combining oral dolasetron with intravenous dexamethasone further improves the antiemetic efficacy of dolasetron. With a number-needed-to treat of about 6 the additional benefit might be considered clinically relevant. PMID- 12624697 TI - [Emergency from anesthesia in small children. From laryngospasm to prolonged apnea]. AB - Postoperative laryngospasm during emergence from anaesthesia represents a potentially life-threatening complication. Even if this is successfully overcome using drug therapy, new, serious problems may develop. We report the case of a 3 1/2 -year-old boy of African descent weighing 15 kg who developed a laryngospasm during emergence from anaesthesia. Because the airway obstruction could not be controlled by deepening the anaesthesia again and administering anti-obstructive drugs, the boy was given 15 mg succinylcholine. Thereafter prolonged apnea developed such that the patient had to be admitted to the pediatric intensive care unit. The child was extubated 6 h later and the further course was normal so that he could be released from the hospital the following day. Further diagnostic study revealed a dibucaine-sensitive, fluoride-resistant pseudocholinesterase in the plasma, which is a rare form of atypical pseudocholinesterase, explaining the prolonged arousal phase after the administration of succinylcholine. Three significant aspects of this case are discussed: 1. risk factors and treatment of perioperative airway obstruction 2. factors and treatment of prolonged apnea, and 3. delayed arousal reactions and their management in an outpatient setting. PMID- 12624698 TI - [Delayed negative pressure pulmonary edema]. AB - A young athletic male adult (smoker) developed a pulmonary edema 30 min after the end of anaesthesia. Extubation was complicated by a laryngospasm. After artificial ventilation for 12 h the patient recovered completely.A negative pressure pulmonary edema (NPPE) develops after deep inspiratory efforts with an occluded airway. Such a maneuver leads to negative intrapleural pressures of -50 to -100 mmHg. This pressure gradient causes damage to the pulmonary capillaries, a transcapillary volume shift into the interstitium,and hemodynamic changes that increase the intrapulmonary blood volume. As a NPPE may occur with a delay of up to 1 h it is crucial to provide adequate monitoring for patients at risk. Symptomatic therapy usually leads to complete recovery within several hours. PMID- 12624699 TI - [Peripartum cardiomyopathy. Perioperative anaesthesiological management of a rare complication of pregnancy]. AB - Peripartum cardiomyopathy is a rare disorder with an incidence from 1:3,000 to 1:15,000 live births and thus not often described in the anaesthesiology literature. The etiology of this disease is still not known but the symptoms are similar to idiopathic dilated cardiomyopathy. Echocardiographic findings show a dilatation of the left ventricle in addition to abnormal wall motion with a severe reduction of the cardiac function. Despite the rarity of this disorder, the anaesthesiologist or ICU physician should consider peripartal cardiomyopathy as a differential diagnosis to ensure an adequate perioperative management. There seems to be an increased incidence in pregnant women who are elderly (age >30 years),who have a history of gestosis/hypertension,have a gemini pregnancy or are of black origin. The prognosis depends on the recovery of the left ventricular contractility within the first 6 months after onset of the disease. The mortality rate is reported to vary between 25% and 50%. Heart transplantation is regarded as the last resort which has successfully been performed with several patients. This case describes the perioperative management of a 32-year-old women with peripartum cardiomyopathy. PMID- 12624700 TI - [Selective digestive tract decontamination in intensive care medicine. Fundamentals and current evaluation]. AB - Selective digestive tract decontamination (SDD) is a method where topical non absorbable antibiotics are applied to the oropharynx and stomach which primarily is aimed at the prevention of ventilator-associated pneumonia. The rationale for SDD is that ventilator associated pneumonia usually originates from the patients'own oropharyngeal microflora. SDD is also used for the prevention of gut derived infections in acute necrotizing pancreatitis and in liver transplantation. Despite numerous clinical trials and several meta-analyses, SDD is still a controversial topic. It is now commonly accepted that the incidence of pneumonia is reduced,however, the concept of using topical antibiotics has its inherent limitations and the best results have been obtained by combination with a short course of intravenous antibiotics. Several issues surrounding the notorious difficulties in establishing the diagnosis of ventilator-associated pneumonia especially in the presence of antibiotics are an on-going matter of debate.Furthermore, pneumonia is the leading cause of death from nosocomial infections and its prevention was not adequately followed by reduced mortality in most individual trials, however, a benefit was suggested by recalculation of data in meta-analyses. Patients are not well defined by their need for ICU admission and mechanical ventilation and the attributable mortality of infections depends more on the type and severity of the underlying diseases. Recently published trials substantially improved our understanding as to which patients may derive most benefit from SDD.Currently, it seems that an improved survival can be achieved in surgical and trauma patients with severe but salvageable diseases, which might be classified e.g.by calculation of APACHE-II scores on admission.However, the most important drawback of SDD is the development of resistance and an increased selection pressure towards Gram-positive pathogens, especially in institutions with endemic multi-resistant microorganisms.Thus, it appears that "selective" must not only be interpreted as selective suppression of pathogenic bacteria but rather as selection of appropriate groups of patients with respect to underlying diseases and severity of illness. Furthermore, it means selection of ICUs where the endemic resistance patterns might allow the use of SDD at a relatively low risk for selection of resistant microorganisms, which is still the major concern associated with SDD. PMID- 12624701 TI - [Accessible price lists at the anaesthesiologist's workplace enhance cost consciousness as a part of process and cost optimization]. AB - The imminent introduction of the DRG (diagnosis-related-group) system is putting hospitals in Germany under considerable pressure. This requires that personnel are efficiently allocated by optimizing organizational procedures and that the limited resources be distributed in a cost-effective manner. One prerequisite for this is a marked cost-consciousness on the part of those who "incur costs" in providing a service. To increase the awareness of costs in clinical physicians, the cost structures must be transparent. In order to achieve this goal, a project was initiated at the Department of Anaesthesiology at the University Hospital of Heidelberg, which aimed to enhance the cost-consciousness of the staff by making price lists available to anaesthesiologists at the workplace. In addition to the price lists, the 25 most expensive medications and medical products were added as an ABC analysis. The departmental staff was interviewed by questionnaire as to whether this project was reasonable. After 1 year the interview was repeated. The results of the questionnaire showed that in the opinion of the staff, price lists are an effective tool, as cost-consciousness on the part of clinical physicians can be enhanced by making price structures transparent. This is a major prerequisite for individual motivation in the cost-effective management. Although the ABC analyses demonstrate no long-term effect of the price-transparency on the cost structures, the staff showed increased cost-consciousness and individual motivation for economic tasks. PMID- 12624702 TI - [High resolution fluorescence microscopy in combination with mathematical modelling. First evidence of sub-cellular anesthetic effects on Ca2+ sparks in situ]. AB - Volatile anesthetics used in daily clinical routine, are associated with a rare but life-threatening disease, malignant hyperthermia. To date it is well known that, with the exception of xenon and nitrous oxide, all volatile anesthetics have the potential to trigger calcium (Ca(2+)) release from the sarcoplasmic reticulum, thereby influencing the Ca(2+) homeostasis in muscle fibers. The effects of volatile anesthetics have been previously studied by recording Ca(2+) activated force transients in muscle fibers and by quantifying the effects on isolated intracellular Ca(2+)-release channels (ryanodin receptors). The use of high resolution fluorescence microscopy methods in combination with spatio temporal mathematical models allows the effects of volatile anesthetics on functional clusters of ryanodin receptors in mammalian skeletal muscle fibers to be studied in situ for the first time.Thus, the analysis of cellular Ca(2+) activated force production and single channel properties in conjunction with mathematical models allows the quantification of the effects of volatile anesthetics on Ca(2+)-release in the natural physiological environment on the basis of the underlying molecular architecture. In addition to the basic understanding of alterations in the Ca(2+) homeostasis induced by volatile anesthetics in muscle and nerve cells, the results are also of direct clinical importance for the understanding of the pathogenesis of malignant hyperthermia,where ryanodin receptor mutations are currently thought to result in an increased Ca(2+) release under the influence of volatile anesthetics. PMID- 12624703 TI - [Direct transfusion of surgical wound blood]. PMID- 12624704 TI - [Anaesthetic concepts in vascular surgery. The value of local anaesthetic procedures]. PMID- 12624705 TI - The treatment of ankylosing spondylitis. AB - Ankylosing spondylitis is an inflammatory disorder affecting the axial skeleton and periphery. Symptoms can often be debilitating. Current therapy for the disease include physical therapy, non-steroidal anti-inflammatory drugs (NSAIDS, anti-rheumatic disease modifying drugs (DMARDS), and the newly developed biologic agents targeting tumor necrosis factor alpha (TNF-alpha). This paper will provide a comprehensive review of these treatments which focusing on evidence based medicine for the daily clinical practice of rheumatology. PMID- 12624706 TI - Endothelin-1 concentration in plasma is increased after jogging but decreased after cycling in healthy men. AB - Endothelin-1 plays an important role as a paracrine factor in the regulation of regional blood flow. Plasma levels may represent the net result of spill-over from local stimulation/release and elimination of endothelin-1. In order to compare changes in the concentration of endothelin-1 in the plasma of subjects performing different types of sports exercises we measured immunoreactive endothelin-1 in healthy volunteers ( n=20) performing in random order jogging on a treadmill and cycling on a bicycle ergometer, for 30 min each. Plasma immunoreactive endothelin-1 increased significantly after jogging (2.13+/-0.8 pg/ml versus 2.6+/-0.8 pg/ml, before and after exercise, respectively, P<0.03), whereas it decreased after cycling (2.45+/-0.76 pg/ml versus 2.25+/-0.9 pg/ml, before and after exercise, respectively, P<0.03). We suggest that microtraumatizing effects on the endothelial lining are more pronounced during jogging than during cycling, resulting in a greater increase in plasma endothelin 1, which is too high to be immediately eliminated by the lung despite exercise induced enhanced pulmonary perfusion. In contrast, similarly enhanced lung perfusion together with a relatively lower stimulation of endothelin-1 compared with jogging, may explain the net decrease in plasma after cycling. PMID- 12624707 TI - An increase in lipoprotein oxidation and endogenous lipid peroxides in serum of obese women. AB - Endogenous malondialdehyde and diene conjugate levels, the susceptibility of apolipoprotein B-containing lipoproteins to copper-induced lipid peroxidation, and antibody titer against oxidized low-density lipoproteins were increased, but serum antioxidant activity was unchanged in obese women. Serum cholesterol, low density lipoproteincholesterol, and trigliceride levels were also elevated, but high-density lipoprotein-cholesterol levels remained unchanged in obese women. In vitro, oxidation of apolipoprotein B-containing lipoproteins and levels of antibody against oxidized low-density lipoprotein correlated with body mass index, serum total cholesterol, and low-density lipoproteincholesterol levels in obese women. These results indicate that obesity is associated with increases in endogenous lipid peroxides, oxidation of low-density lipoproteins, and lipids in serum. PMID- 12624708 TI - Association of apolipoprotein E gene polymorphism with essential hypertension and its complications. AB - The clinical significance of the apolipoprotein E genotype in patients with hypertension has been a subject of debate. We enrolled 94 patients with hypertension and 102 healthy controls in this study and determined their plasma levels of triglyceride, total cholesterol, high- and low-density lipoprotein cholesterol, apolipoprotein AI, and apolipoprotein B. The apolipoprotein E genotypes were identified by polymerase chain reaction, restriction fragment length polymorphism, and polyacrylamide gel electrophoresis. Apolipoprotein E3/4 genotype and set membership, vertical bar on horizontal stroke 4 allele frequencies in the hypertensive group were higher than in controls. In hypertensive patients with apolipoprotein E3/4 and E4/4 genotypes, systolic blood pressure was significantly higher than in those with apolipoprotein E2/3 or E3/3 genotypes. Meanwhile, the plasma levels of total cholesterol, low-density lipoprotein-cholesterol, and apolipoprotein B were higher in hypertensive patients with the.4 allele than the set membership, vertical bar on horizontal stroke 2 or set membership, vertical bar on horizontal stroke 3 allele. The echographic measurements of carotid artery intimal-medial thickness showed increasing values from set membership, vertical bar on horizontal stroke 2 to set membership, vertical bar on horizontal stroke 4 allele carriers in the hypertensive group. Analysis of variance showed that the carotid intimal-medial thickness was significantly greater in hypertensive patients with set membership, vertical bar on horizontal stroke 4 alleles compared with set membership, vertical bar on horizontal stroke 2 or set membership, vertical bar on horizontal stroke 3 alleles. Our data show an association between apolipoprotein E genotype and hypertension and support the hypothesis that the apolipoprotein set membership, vertical bar on horizontal stroke 4 allele is a susceptibility locus for systolic hypertension and carotid artery atherosclerosis. PMID- 12624709 TI - Effects of cilostazol on serum lipid concentrations and plasma fatty acid composition in type 2 diabetic patients with peripheral vascular disease. AB - Cilostazol is an anti-thrombotic and vasodilating agent, reported to have both anti-thrombotic and cerebral vasodilating effects. We investigated the effects of cilostazol on serum lipid concentrations and plasma fatty acid composition in type 2 diabetic patients with peripheral vascular disease. The serum concentrations of total cholesterol, triglycerides, high-density lipoprotein cholesterol, lipoprotein (a), remnant-like particles-cholesterol, apolipoproteins, and plasma fatty acid composition were measured in 17 diabetic patients with peripheral vascular disease before and 1, 3, and 6 months after administration of cilostazol (200 mg/day). Serum triglyceride concentrations were significantly decreased after cilostazol (from 1.31+/-0.17 mmol/l to 0.86+/-0.07 mmol/l at 6 months, P<0.01). Plasma docosahexaenoic acid levels were significantly increased after cilostazol (4.11+/-0.26% to 4.94+/-0.26% at 6 months, P<0.01). Our findings show that cilostazol can induce some beneficial changes in serum lipid profile and plasma fatty acid composition. PMID- 12624710 TI - Fuzzy logic-based tumor marker profiles improved sensitivity of the detection of progression in small-cell lung cancer patients. AB - Tumor markers were used for disease monitoring in small-cell lung cancer patients. The aim of this study was to improve diagnostic efficiency in the detection of tumor progression in small-cell lung cancer patients by using fuzzy logic modeling in combination with a tumor marker panel (NSE, ProGRP, Tumor M2 PK, CYFRA 21-1, and CEA). Thirty-three consecutive small-cell lung cancer patients were included in a prospective study. The changes in blood levels of tumor markers and their analysis by fuzzy logic modeling were compared with the clinical evaluation of response versus non-response to therapy. Clinical monitoring was performed according to the standard criteria of the WHO. Tumor M2 PK was measured in plasma with an ELISA, all other markers were measured in sera. At 90% specificity, clinically detected tumor progression was found by the best single marker, NSE, in 32% of all cases. A fuzzy logic rule-based system employing a tumor marker panel increased the sensitivity significantly (P>0.0001) in small-cell carcinomas to 67% with the threemarker combination NSE/ProGRP/Tumor M2-PK and to 56% with the best two-marker combination ProGRP/Tumor M2-PK, respectively. An improvement of sensitivity was also observed using the two marker combination of ProGRP/NSE (sensitivity 49%) or NSE/Tumor M2-PK (sensitivity 52%). The fuzzy classifier was able to detect a higher rate of progression in small-cell lung cancer patients compared with the multiple logistic regression analysis using the marker combination NSE/ProGRP/Tumor M2-PK (sensitivity 44%; AUC=0.76). With the fuzzy logic method and different tumor marker panels (NSE, ProGRP and Tumor M2-PK), a new diagnostic tool for the detection of progression in patients with small-cell lung cancer is available. PMID- 12624711 TI - High levels of serum soluble CD27 correlated with renal dysfunction. AB - CD27 is expressed on lymphocytes, but also on renal tubules. On the damaged renal tubules, CD27 is cleaved into soluble CD27 (sCD27) and cytoplasmic tail CD27. Renal tubule apoptosis is induced by the CD27 ligand, Siva, binding to the truncated tail CD27. Theoretically, serum sCD27 should be a marker of renal tubule apoptosis. Serum sCD27 levels were measured by ELISA in 274 patients at University Hospital School of Medicine, Japan. Among 73 males and 63 females with high plasma creatinine levels, 68 (93%) males and 51 (81%) females showed high serum sCD27 values (>or=500 U/ml). In the 42 males with plasma creatinine levels of 1.18-5.00 mg/dl and the 55 females with plasma creatinine levels of 0.83-4.65 mg/dl, the correlation coefficients between plasma creatinine and sCD27 were 0.605 and 0.469, respectively. SCD27 was a more sensitive marker of renal tubule apoptosis than plasma creatinine. Of 138 patients with normal plasma creatinine, 42 showed high serum sCD27 levels (>or=500 U/ml). Among the 42 patients, 19 patients had high blood urea nitrogen and 11 patients showed plasma creatinine elevations 3-11 days later. Nine patients with hemolysis and 39 patients with aneurysms had clearly higher sCD27 levels of 1633+/-483 U/ml and 905+/-437 U/ml, respectively, than normal values of 215+/-57 U/ml ( n=27) ( P value >or=0.001). Vasodilators, hypotensors, diuretics, and drugs for gout, arrhythmia, thrombosis, and diabetes were prescribed in 40%, 37%, 33%, 13%, 8%, 8%, and 5% of the 78 patients analyzed with high sCD27 levels (>or=500 U/ml). In total, 59 (76%) patients were prescribed at least one of the drugs. In conclusion, serum sCD27 was a more sensitive indicator of renal tubule apoptosis than plasma creatinine, especially during the early stage of apoptosis. Tubule apoptosis with high serum sCD27 levels was found in patients with glomerular vascular injuries. PMID- 12624712 TI - Study of phagocytosis of senescent erythrocytes in young and elderly individuals. AB - There is a decrease in the percentage contribution of a heavy density fraction of red blood cells to whole blood with increasing age. The aim of this study was to investigate in the young and elderly the interaction between monocytes and different erythrocyte suspensions: senescent red blood cells, erythrocytes stored with or without serum, and desialylated red blood cells. The results obtained with senescent red blood cells and erythrocytes stored with serum show the involvement of autologous IgG in the selective removal of erythrocytes. These values were higher in elderly individuals, indicating that this process increases with age. Our observation suggest that desialylation is not involved in the increased removal of erythrocytes observed in elderly individuals. PMID- 12624713 TI - Treatment of migraine with aura: comments and perspectives. AB - Migraine with aura (MwA) is a primary headache that affects about 30% of migraine sufferers. The main questions for the physician caring for the patient who has MwA are: when to use preventive medications, what medications to use in acute and preventive treatment, and whether the aura should be treated. The aim of this paper is to review the various therapeutic options for MwA proposed in the current literature and to evaluate their efficacy. PMID- 12624714 TI - Investigation of fine-motor disturbances in Wilson's disease. AB - Patients suffering from Wilson's disease (WD) can be divided into two main subgroups: neurologic and nonneurologic WD. We measured passive and active fine motor abilities of 37 WD patients and 24 randomly selected volunteers. The measurement was based on a standardized test set in a defined environment for detection of disturbed finemotor control. The set contains 5 tests comprising rest tremor, postural tremor, target tapping, forefinger tapping and spiral painting, reflecting different aspects of movement disorders. The tests showed significant differences between neurologic WD and volunteers, especially for tasks defining active control. In neurologic WD we found no differences between subgroups whereas for non-neurologic WD we often detected slight movement disorders. The detected movement disorders cam be interpreted as persistent disorders after long-term therapy. PMID- 12624715 TI - A study of arm movements in Huntington's disease under visually controlled and blindfolded conditions. AB - The so-called bradykinesia of Huntington's disease (HD) seems not due to reduced movement speed alone but may also be task-dependent. We therefore investigated the influence of visual control on the ability of HD patients to perform a motor task. Ten HD patients, never treated with neuroleptic drugs and with mild functional impairment in activities of daily living, performed the task blindfolded and not blindfolded, as did 10 age- and education-matched healthy controls. The task was to use the dominant hand to trace out the contours of a 20 x 20 cm square in a clockwise direction, pausing at each corner. The square was marked on the table at which the subject sat. Accuracy was stressed rather than speed. A videocamerabased system recorded movement trajectories, from which kinematic and error parameters were derived. Patients and controls moved at comparable speeds but patients took longer to complete the task due to more curvilinear and hence longer trajectories. Patients spent more time in the deceleration phase of the movement, and in the blindfold condition had more variable movements as indicated by greater error variability scores. Correlation analysis showed that kinematic parameters in patients did not correlate with involuntary movement scores. These findings indicate that abnormalities of motor control are present in HD when movement accuracy (and not velocity) is required. HD patients are more dependent on visual control than normal subjects. PMID- 12624716 TI - Italian version of the Epworth sleepiness scale: external validity. AB - We assessed the validity of an Italian language version of the Epworth sleepiness scale (ESS). The translated ESS was compared to the multiple sleep latency test (MSLT), considered the gold standard for the diagnosis of excessive daytime sleepiness (EDS). Within the context of a multicentric national study on narcolepsy (Gruppo Italiano Narcolessia Studio Epidemiologico Nazionale, GINSEN) involving 17 Italian sleep centres, we compared the two diagnostic tests on 91 prospectively recruited subjects with suspected EDS (34 with narcolepsy, 16 with obstructive sleep apnea syndrome, 19 with idiopathic hypersomnia, and 22 with other sleep, neurologic or psychiatric disorders). ESS scores were inversely correlated with mean sleep latency values, as measured with MSLT (rho = -0.31, p<0.01). ESS cut-off scores with best sensitivity and specificity were 12 and 17. For the 5-min MSLT cut-off, sensitivity was 87% and 47% respectively; specificity 39% and 74%. For the 8-min MSLT cut-off, sensitivity was 84% and 49%; specificity 50% and 88%. The Italian version of the ESS is an easy-to-use form useful for preliminary screening of daytime sleepiness level in specialist settings. PMID- 12624717 TI - A genetic association study of migraine with dopamine receptor 4, dopamine transporter and dopamine-beta-hydroxylase genes. AB - We assessed the role of some dopamine metabolism genes in the genetic susceptibility to migraine. We performed an association study using three functional polymorphisms: a 48-base-pair (bp) tandem repeat in the D4 dopamine receptor gene ( DRD4), a 40-bp tandem repeat in the dopamine transporter gene ( DAT) and a dinucleotide repeat in the dopamine beta-hydroxylase ( DBH) gene. Allelic and genotypic frequencies for each polymorphism were assayed in two migraine populations (93 individuals with migraine with aura (MA) and 101 with migraine without aura (MO)) and were compared with those in a control group (117 individuals). No significant differences were found between control and migraine groups for DAT and DBH polymorphisms. Instead, the distribution of alleles for the DRD4 gene in the MO group was significantly different from those in both MA and control groups, with the shortest and longest alleles being less frequent in MO. Our data indicate that MO, but not MA, shows significant genetic association with DRD4. PMID- 12624718 TI - Intracranial stimulation of the facial nerve: normative values with magnetic coil in 240 nerves. AB - This study was aimed at defining normative values of latency and amplitude of the compound muscle action potential (CMAP) from the orbicularis oculi muscle, obtained by transcranial magnetic stimulation. We evaluated the dependence of latency and amplitude on the age of the subjects, in order to calculate cut-off values for each age decade. A total of 120 healthy normal subjects, aged 15-78 years, were studied. CMAP from the orbicularis oculi muscle, pars inferior, was excited by means of transcranial magnetic stimulation of the homolateral scalp. A suitable mathematical model was developed to evaluate the mean variation of the latency and amplitude of CMAP for each age decade in the studied population. Mean values were, respectively, 4.62+/-0.40 ms for latency and 4.17+/-0.72 mV for amplitude. We found an increase in latency of about 0.12 ms for every ten years, that became 0.15 ms if referring only to subjects over 20 years, and a decrease in amplitude of 0.18 mV each ten years. Normality interval values for the latency and amplitude were calculated for each age decade. Our data, obtained in a representative population for range of age of evaluated subjects, provide normality values and variability coefficients useful for future confronting works. PMID- 12624719 TI - Acute disseminated encephalomyelitis after bee sting. AB - Acute disseminated encephalomyelitis (ADEM) usually follows a viral infection or an immunization and is thought to be an immunomediated disease. We describe a patient with ADEM after multiple yellow jacket bee stings. The patient recovered after treatment with a high dose of methylprednisolone. Although the pathologic mechanism exact remains unclear, potential cross-reactivity between bee toxins and the central nervous system myelin could induce demyelination. ADEM should be considered a rare complication of bee stings. PMID- 12624720 TI - Crossed aphasia with left spatial neglect and visual imperception: a case report. AB - A 64-year-old right-handed woman with no left-handers in the family developed aphasia associated with moderate left hemiparesis and dense left homonymous hemianopia following rupture of a right middle cerebral artery aneurysm and subsequent selective surgery confined to the right hemisphere. Severe left spatial neglect and constructional apraxia were also present. The patient was an achondroplasic dwarf whose previous medical and neurological history was otherwise unremarkable. Computed tomography of the brain showed a large right temporo-insulofrontoparietal lesion. Language and nonverbal cognitive functions were assessed after 2 and 6 months, and then four years later. A reportedly overall language disruption in the acute period evolved into Wernicke's aphasia and then into a mild form of conduction aphasia. The associated left spatial neglect eventually shrank to a minimum. The patient never had clinically detectable visual agnosia, but on specific tests of visual recognition and perception some impairment was found four years after onset. The left hemiparesis disappeared in time while the left hemianopia persisted. This case is a convincing example of an entirely righthanded person in whom both linguistic and visuospatial functions are represented in the right hemisphere. PMID- 12624721 TI - Physical map and haplotype analysis of 16q-linked autosomal dominant cerebellar ataxia (ADCA) type III in Japan. AB - Autosomal dominant cerebellar ataxia (ADCA) is a group of heterogeneous neurodegenerative disorders. We previously mapped a gene locus for ADCA with pure cerebellar syndrome (ADCA type III) to a 3-cM region in chromosome 16q, and found a common haplotype among affected individuals. This region was exactly within the locus for another ADCA, spinocerebellar ataxia type 4 (SCA4). To identify the gene causing 16q-linked ADCA type III, we constructed a contig with 38 bacterial artificial chromosome clones between D16S3043 and D16S3095. The size of this contig was estimated to be 4.8 Mb. We found more than 500 nucleotide tandem repeats, including 9 CAG/CTG repeats in this candidate region, although none of the 94 tandem repeats analyzed were expanded in affected individuals. However, we found 11 new polymorphic markers, giving 22 markers spanning the candidate region. By typing these markers on eight Japanese families with ADCA type III, including two new families, we found that a common "founder" haplotype is seen in a more restricted 3.8-Mb region, spanning markers GGAA05 and D16S3095. We present here a newly refined critical interval of 16q-ADCA type III/SCA4. Data of 11 new DNA markers on 16q22.1 would also be useful for other research of genes mapped to this region. PMID- 12624722 TI - Nonsyndromic sensorineural deafness associated with the A1555G mutation in the mitochondrial small subunit ribosomal RNA in a Balinese family. AB - Sensorineural deafness associated with increased sensitivity to aminoglycoside antibiotics as the consequence of an A1555G mutation in the mitochondrial DNA (mtDNA) in a highly conserved region of the small (12S) rRNA gene has been reported in Caucasian, Chinese, and Japanese individuals. We report here a large family of Balinese Indonesian origin with progressive/congenital sensorineural deafness who carry the A1555G mutation. The pedigree shows a generally maternal inheritance pattern with some exceptions, which is the result of an unusual multiple entry of the mutation into the pedigree. A complete mtDNA genome sequence from three Balinese individuals revealed a relatively large number of single- nucleotide polymorphisms (20) not previously reported, and confirmed the genetic distance of Southeast Asian populations from those of Caucasians and Japanese. The biochemical expression of the A1555G mutation under the influence of this mtDNA background was investigated. Examination of respiratory enzyme activities showed a significant decrease in respiratory complex I activity, particularly in symptomatic family members. PMID- 12624723 TI - Mutation analysis of the ALD gene in seven Japanese families with X-linked adrenoleukodystrophy. AB - The childhood cerebral form of X-linked adrenoleukodystrophy (X-ALD) is a severe congenital metabolic disease without a definite effective therapy except for hematopoietic stem cell transplantation in the appropriate disease stage. Seven Japanese families with X-ALD were analyzed for mutations in the ALD gene ( ALD). Of the seven families, three were referred to us for prenatal diagnosis, four for carrier detection, and three for confirmation diagnosis of patients. By nucleotide sequencing and/or restriction analysis, all the subjects to be examined were successfully diagnosed. Six different missense mutations in ALD were identified. There was a G-->A substitution (G512S) in two unrelated families, and a G-->A (R617H), a C-->T (R660W), a G-->C (R163P), a C-->T (S606L), or a G-->A (G116E) substitution in each of the other five families. Among the six substitutions, five were those reported previously and the other was a novel mutation. In three families, prenatal diagnosis was carried out after genetic counseling. PMID- 12624724 TI - Evaluation of the diagnostic accuracy of the stop codon (SC) assay for identifying protein-truncating mutations in the BRCA1and BRCA2genes in familial breast cancer. AB - Screening for protein-truncating mutations of the BRCA1 and BRCA2 genes is useful in genetic testing for familial breast cancer because, first, the methods are usually simple and not expensive, and second, the detected mutations indicate pathogenic mutations in general. We evaluated the diagnostic accuracy of the stop codon (SC) assay for detecting protein-truncating mutations in the BRCA1 and BRCA2 genes by comparing the results with DNA sequencing in samples from 29 patients with breast cancer from 24 Japanese families with a history of breast cancer. Protein-truncating mutations were detected in 5 of the 24 families (20.8%; two in the BRCA1 gene and three in the BRCA2 gene). Among the 176 DNA fragments examined using the SC assay, the existence of three protein-truncating mutations (one in the BRCA1 gene and two in the BRCA2gene) was predicted correctly by the assay. Only one reverse transcriptase-polymerase chain reaction fragment was positive for the SC assay but was negative using DNA sequencing. Our study showed clearly that the SC assay is sensitive (3 of 3, 100%) and specific (172 of 173, 99%) for detecting pathogenic protein-truncating mutations in the BRCA1 and BRCA2 genes, and that it could be useful for screening larger populations. PMID- 12624725 TI - The polymorphism of manganese superoxide dismutase is associated with diabetic nephropathy in Japanese type 2 diabetic patients. AB - We evaluated the relationship of an alanine or valine polymorphism at amino acid sequence 16 [Val(16)Ala] of manganese superoxide dismutase (Mn-SOD) with diabetes and diabetic nephropathy in Japanese type 2 diabetic patients. Val(16)Ala genotyping of Mn-SOD was done by polymerase chain reaction-restriction fragment length polymorphism with a restriction enzyme ( Bsaw I) in 478 Japanese type 2 diabetic patients and 261 nondiabetic Japanese healthy subjects. The genotype distribution of diabetic and nondiabetic subjects was then compared, and the association of genotype with diabetic nephropathy was evaluated in the diabetic patients. The allele frequency and genotype of the diabetic patients were not different from those of the healthy nondiabetic subjects. The VV type showed a significantly higher frequency in the diabetic patients with nephropathy than did the AA or VA type [VV type: normoalbuminuria 70.8%, microalbuminuria 84.8% (P = 0.0057), macroalbuminuria 84.1% (P = 0.0128)]. Furthermore, logistic regression analysis showed that this polymorphism is associated with diabetic nephropathy independently (odds ratio = 0.461925, P = 0.03). Accordingly, the Val(16)Ala polymorphism of Mn-SOD may be unrelated to the etiology of type 2 diabetes, but it seems to be associated with diabetic nephropathy in Japanese type 2 diabetic patients. PMID- 12624726 TI - Construction and characterization of a vestibular-specific cDNA library using T7 based RNA amplification. AB - Using a very small amount of inner-ear tissue, we constructed a human vestibular cDNA library by means of T7-based amplification of RNA. This library should allow us to identify genes likely to be involved in auditory and vestibular functions. Here we first report the characterization of the human vestibular cDNA library. Among 506 cDNA clones randomly selected from the vestibular cDNA library, DNA sequences of 301 cDNA clones were identical to those of genes of known function. Twenty-two cDNA clones were considered to be novel because they did not match any cDNA sequences in the public database. The information in our study will provide a valuable resource for identifying several novel genes underlying deafness disorders and vestibular dysfunction. PMID- 12624727 TI - High-resolution SNP map in the 55-kb region containing the selectin gene family on chromosome 1q24-q25. AB - Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis characterized by predominant IgA deposits in glomerular mesangium. By means of a genome-wide case-control association study, we previously demonstrated that eight single-nucleotide polymorphisms (SNPs) within the selectin gene cluster are significantly associated with IgAN. Here we provide more detailed information of variations corresponding to selectin loci, consisting of 88 SNPs and two insertion-deletion polymorphisms in the Japanese population: 27 in 5' flanking regions, 1 in 5' untranslated regions, 6 within coding regions, 46 in introns, 4 within 3' untranslated regions, and 4 in 3' flanking regions. The SNP map presented here will be a useful resource not only for examining the relationships between selectin genotypes and susceptibility to the IgAN phenotype, but also for analyzing gene scans of complex diseases mapped to this local segment on chromosome 1. PMID- 12624728 TI - A locus for congenital preauricular fistula maps to chromosome 8q11.1-q13.3. AB - The incidence of congenital preauricular fistula (CPF) is >1.1% in both Chinese and Caucasians, but it is even higher in Blacks. We mapped the locus for CPF to chromosome 8q11.1-q13.3 by linkage analysis of a family composed of 7 affected and 11 nonaffected members. The two-point LOD score was 2.40, shown by markers D8S285 and D8S1113 at a recombination fraction (theta) of 0.00. Results from three other markers (D8S1110, D8S260, and D8S1136) in the same region further support the linkage. Haplotype analysis for this family confined the locus to within an interval of approximately 26.7 cM, flanked by markers D8S532 and D8S279. A LOD score of <3 is likely due to the limitation of family size. PMID- 12624729 TI - Identification of a novel human angiopoietin-like gene expressed mainly in heart. AB - The angiopoietins are an important family of growth factors specific for vascular endothelium. Most of them bind to the TIE2 receptor and are related to regulation of angiogenesis. During large-scale DNA sequencing of the human fetal brain cDNA library, we cloned a novel human angiopoietin-like cDNA and termed it human angiopoietin-like 5 ( ANGPTL5). Like other members of the angiopoietin family, ANGPTL5-deduced protein also has an N-terminal cleavable signal peptide, a predicted coiled-coil domain, and a fibrinogen-like domain. The search against the human genome database indicated that ANGPTL5 maps to 11q22. Expression analysis of ANGPTL5 shows that it is mainly expressed in adult human heart. PMID- 12624730 TI - Characterization of the MCRred2 form of methyl-coenzyme M reductase: a pulse EPR and ENDOR study. AB - Methyl-coenzyme M reductase (MCR), which catalyses the reduction of methyl coenzyme M (CH(3)-S-CoM) with coenzyme B (H-S-CoB) to CH(4) and CoM-S-S-CoB, contains the nickel porphinoid F430 as prosthetic group. The active enzyme exhibits the Ni(I)-derived axial EPR signal MCR(red1) both in the absence and presence of the substrates. When the enzyme is competitively inhibited by coenzyme M (HS-CoM) the MCR(red1) signal is partially converted into the rhombic EPR signal MCR(red2). To obtain deeper insight into the geometric and electronic structure of the red2 form, pulse EPR and ENDOR spectroscopy at X- and Q-band microwave frequencies was used. Hyperfine interactions of the four pyrrole nitrogens were determined from ENDOR and HYSCORE data, which revealed two sets of nitrogens with hyperfine couplings differing by about a factor of two. In addition, ENDOR data enabled observation of two nearly isotropic (1)H hyperfine interactions. Both the nitrogen and proton data indicate that the substrate analogue coenzyme M is axially coordinated to Ni(I) in the MCR(red2) state. PMID- 12624731 TI - Homocysteine in the context of cobalamin metabolism and deficiency states. AB - It is becoming increasingly clear that serum vitamin B12 (cobalamin) concentration is a dubious indicator of functional B12 status and, in contrast to long-standing convention, correlates poorly with haematological indices. This, in turn, has led to poorly defined reference intervals for serum B12. Patients presenting with neurological disturbance due to B12 deficiency are at risk of not being diagnosed if total reliance is placed on serum B12 levels and haematological parameters. Plasma homocysteine remethylation is uniquely placed at the metabolic end-point of B12 metabolism such that plasma total homocysteine is proving to be a sensitive marker of functional B12 status. Studies also show that plasma homocysteine correlates better with holotranscobalamin than serum B12. It is suggested that clinicians should cease to be guided by surrogate haematological markers when more specific tests of B12 deficiency, such as holotranscobalamin and total homocysteine, exist. These tests demand greater prevalence in routine diagnostic use. PMID- 12624732 TI - New perspectives on the role of amine oxidases in physiopathology. AB - In the paper here presented we summarize some results obtained in our laboratory in the last few years on new structural and functional aspects of some amine oxidases (AOs), which have to be taken into consideration in defining new strategies of controlling the cellular physiopathology. In particular, the ability of Cu-AO purified from vegetal sources or from bovine serum to bind different cellular targets inducing in them conformational as well as chemical modifications are described and the consequences of this interaction on cellular functions are discussed. This is the case of the protective effect of Cu-AO against the damage induced by free radicals, cell enrichment with Cu-AO, induction of cataract and the leukocyte-endothelia interaction. The role of Cu and FAD-amine oxidases related as to the protection or damage of cells is also discussed. In this context the involvement of MAOs in the modulation of the mitochondrial functions and in the induction of apoptosis is described and some aspects of the molecular mechanism of AO inhibition by H(2)O(2) and metronidazole analyzed. PMID- 12624733 TI - Proteomic analysis of the cell envelope fraction of Escherichia coli. AB - We applied proteomics technologies to analyze a membrane preparation of Escherichia coli, wild type strain and of transformants expressing human cytochrome P450s. The proteins were analyzed by two-dimensional electrophoresis and identified by matrix-assisted laser desorption ionization mass spectrometry. The membrane proteins were solubilized with both mild detergents such as CHAPS and strong detergents, such as sodium and lithium dodecyl sulfate, sodium cholate and sodium deoxycholate. In the E. colimembrane fraction, 394 different gene products were identified. Approximately 28% of them were predicted to be integral membrane proteins, of which 100 proteins have been predicted to carry one transmembrane region, ten proteins to carry two, and two proteins to include three transmembrane domains. The remaining are probably membrane-associated and cytosolic proteins. Cytochrome P450s did not enter the immobilized pH gradient strips but were efficiently analyzed in a two-dimensional, two-detergent system. Use of strong solubilizing agents resulted in the detection of about 20 membrane proteins, which were not detected following extraction with mild detergents and chaotropes. The present database is one of the largest for membrane proteins. PMID- 12624734 TI - Chromatographic determination of L- and D-amino acids in plants. AB - Quantities of free L- and D-amino acids (L- and D-AAs) in plants (leaves of coniferous and decidious trees, fleshy fruits, leaf blades of fodder grasses, and seeds and seedlings of edible legumes) were determined. Amino acid (AA) enantiomers were converted into diastereomers using pre-column derivatization with o-phthaldialdehyde together with N-isobutyryl-L(or D)-cysteine followed by separation of the resulting fluorescent isoindol derivatives on an octadecylsilyl stationary phase using high-performance liquid chromatography. Relative amounts of D-AAs were also determined by enantioselective gas chromatography-mass spectrometry on Chirasil-L-Val. Free D-AAs acids in the range of about 0.2% up to 8% relative to the corresponding L-AAs acids were found in plants. D-Asp, D-Asn, D-Glu, D-Gln, D-Ser and D-Ala could be detected in most of the plants, and D-Pro, D-Val, D-Leu and D-Lys in certain plants. As D-AAs were detected in gymnosperms as well as mono- and dicotyledonous angiosperms of major plant families it is concluded that free D-AAs in the low percentage range are principle constituents of plants. PMID- 12624735 TI - Effects of human growth hormone (hrGH) treatment on amine metabolism in rats subjected to extensive small bowel resection. AB - The effect of human recombinant growth hormone (hrGH) on intestinal adaptation in rats subjected to massive small bowel resection has been followed by monitoring changes in the tissue polyamine system and in red blood cell (RBC) polyamine levels. In parallel, the activities of monoamine oxidase A and B and diamine oxidase, the enzymes that catalyse one of the major routes of biogenic amine metabolism, oxidative deamination, were also examined. The results suggest that whilst hrGH treatment accelerates adaptive intestinal hyperplasia evoked by the resection, it has no significant effect on RBC polyamine level or gut mucosal DNA concentration as measured 3 weeks post surgery. hrGH treated operated rats exhibited significantly lower amine oxidase activities which implies that GH may alter biogenic amine systems. PMID- 12624736 TI - Polyamine metabolism in primary human colon adenocarcinoma cells (SW480) and their lymph node metastatic derivatives (SW620). AB - The natural polyamines are multifunctional constituents of all eucaryotic cells. The objective of this work was to compare aspects of polyamine metabolism in two related cell lines with the idea to investigate whether metabolic differences can be attributed to functional differences of the cells. The human colon carcinoma derived cell lines SW480 and SW620 were chosen as models. SW480 cells were isolated from the primary tumour, SW620 cells from a lymph node of the same patient. SW620 cells grow faster, and the key regulatory enzymes of polyamine biosynthesis (ODC and AdoMetDC) are more active in the metastatic cells. Moreover, their ability to accumulate polyamines from the environment is more important than of SW480 cells. Likewise polyamine concentrations were markedly higher in SW620 cells, although they are much smaller than SW480 cells, and have a particularly small cytoplasmic space. Both cell lines show a striking diminution of ODC and AdoMetDC activities and changes in the polyamine patterns at the transition from exponential to non-exponential growth--most probably as a consequence of high cell density. Depletion of putrescine and spermidine due to inactivation of ODC by DFMO causes accumulation of cells in G1, and a proportional decrease of S-phase cells in both cell lines. Based on morphologic and other criteria SW480 and SW620 cells were typified as poorly differentiated. In agreement with their low grade of differentiation they exhibit a low alkaline phosphatase activity. However, the time-dependent decrease of alkaline phosphatase is not typical of differentiation patterns of other adenocarcinoma derived cell lines or of normal enterocytes. The high capacity of de novo polyamine biosynthesis and of polyamine uptake is presumably a prerequisite for the rapid growth and invasiveness. The fact that these properties were more accentuated in the case of SW620 cells and paralleled enhanced metastatic properties indicate relationships between basic parameters of polyamine metabolism and malignancy. PMID- 12624737 TI - Degradation of tryptophan and related indolic compounds by ruminal bacteria, protozoa and their mixture in vitro. AB - In vitro experiments were conducted to examine the degradation of d- and l isomers of tryptophan (Trp) and 10 related indolic compounds by mixed rumen bacteria (B), protozoa (P) and a combination of the two (BP). The analyses were carried out by HPLC. d-Trp (1.0 mM) was not degraded by rumen microorganisms during the 24-h incubation period. The net degradation of 1 mM l-Trp was 46.5%, 8.7% and 80.0% by B, P and BP suspensions, respectively. Trp was degraded into indoleacetic acid, indolelactic acid and indole by rumen bacteria and protozoa, and into skatole, p-cresol and indolepropionic acid by rumen bacteria only. Of them, indoleacetic acid was the major product of Trp found in B (15.4%) and P (3.1%), and skatole in BP (43.2%). This is the first report of the production of indolelactic acid and p-cresol from Trp by rumen microbes. Starch, d-glucose, salinomycin and monensin inhibited the production of skatole and indole from Trp, and skatole from indoleacetic acid by rumen bacteria. PMID- 12624738 TI - Branched chain amino acids as source of specific branched chain volatile fatty acids during the fermentation process of fish sauce. AB - The source of the formation of branched chain volatile fatty acids (VFA) in fish sauce was investigated. Certain branched VFA were derived from the degradation of specific amino acids as iso-butyric acid from valine and iso-valeric acid from leucine. Short and long straight chain VFA were significantly higher in the linoleic acid added sample than in the control but did not significantly bring changes to the branched chain VFA. It is suggested that straight chain VFA developed from fish fats. Alanine and isoleucine did not have a clear influence on the production of volatile fatty acids. PMID- 12624739 TI - Co-variation of plasma sodium, taurine and other amino acid levels in critical illness. AB - This study investigates the relationship between changes in plasma sodium and changes in amino acid levels in a patient with post-traumatic sepsis and prolonged critical illness. Ninety-two consecutive measurements were performed at regular intervals over a period of many weeks; these consisted in the determination of full amino-acidograms, plasma sodium and complementary variables. A unique, highly significant inverse correlation between taurine and plasma sodium was found (r(2) = 0.48, p < 0.001). All other amino acids were unrelated, or much more weakly related, to sodium. Taurine was also strongly and directly related to phosphoethanolamine, glutamate and aspartate. Changes in sodium and in levels of these amino acids explained up to 86% of the variability of taurine. Besides, levels of these amino acids maintained a high degree of co variation, remaining reciprocally related one to each other, directly, with r(2) ranging between 0.33 and 0.59 (p < 0.001 for all). There were similar findings for beta-alanine, which however was measured inconsistently. These data provide gross clinical evidence of a specific link binding plasma sodium and taurine levels, and may be consistent with occurrence of opposite and interdependent shifts of sodium and taurine between intravascular and extravascular space, to maintain osmoregulation. Co-variation of taurine with the other amino acids may be related to the same phenomenon, and/or to similarities in transport systems and chemical structure, or true metabolic interactions. PMID- 12624740 TI - Mitochondrial nitric oxide inhibits ATP synthesis. Effect of free calcium in rat heart. AB - Nitric oxide is a small potentially toxic molecule and a diatomic free radical. We report the interaction of L-arginine, oxygen and calcium with the synthesis of nitric oxide in heart mitochondria. Nitric oxide synthesis is increased in broken rat heart mitochondria compared with intact and permeabilized mitochondria. Intact mitochondria subjected to hypoxia-reoxygenation conditions accumulated nitric oxide that inhibits oxygen consumption and ATP synthesis. ATPase activity is not affected during this augment of nitric oxide. Physiological free calcium concentrations protected mitochondria from the damage caused by the accumulation of nitric oxide. Higher concentrations of the divalent cation increase the damage exerted by nitric oxide. PMID- 12624741 TI - Alterations in hepatic metabolism of sulfur-containing amino acids induced by ethanol in rats. AB - Alterations in hepatic metabolism of S-amino acids were monitored over one week in male rats treated with a single dose of ethanol (3 g/kg, ip). Methionine and S adenosylhomocysteine concentrations were increased rapidly, but S adenosylmethionine, cysteine, and glutathione (GSH) decreased following ethanol administration. Activities of methionine adenosyltransferase, cystathionine gamma lyase and cystathionine beta-synthase were all inhibited. gamma-Glutamylcysteine synthetase activity was increased from t = 8 hr, but GSH level did not return to control for 24 hr. Hepatic hypotaurine and taurine levels were elevated immediately, but reduced below control in 18 hr. Changes in serum and urinary taurine levels were consistent with results observed in liver. Cysteine dioxygenase activity was increased rapidly, but declined from t = 24 hr. The results show that a single dose of ethanol induces profound changes in hepatic S amino acid metabolism, some of which persist for several days. Ethanol not only inhibits the cysteine synthesis but suppresses the cysteine availability further by enhancing its irreversible catabolism to taurine, which would play a significant role in the depletion of hepatic GSH. PMID- 12624742 TI - Protein levels of genes encoded on chromosome 21 in fetal Down syndrome brain: challenging the gene dosage effect hypothesis (Part I). AB - Down syndrome (DS) is the most significant genetic disorder with mental retardation and is caused by trisomy 21. The phenotype of DS is thought to result from overexpression of a gene(s) located on the triplicated chromosome (region). An increasing body of evidence that challenge this "gene dosage effect" hypothesis, however, has been reported indicating that this hypothesis still remains to be elucidated. The availability of the complete sequence of genes on chromosome 21 could have an immediate impact on DS research, but no conclusions can be drawn from nucleic acid levels. This made us evaluate protein levels of six proteins, gene products, encoded on chromosome 21 (T-cell lymphoma invasion and metastasis inducing Tiam1 protein, holocarboxylase synthetase, human interferon-regulated resistance GTP-binding protein MxA, Pbx regulating protein 1, autoimmune regulator, and pericentrin) in fetal cortex from DS and controls at 18-19 weeks of gestational age using Western blot technique. None of the investigated proteins showed overexpression in DS compared to controls. Our present data showing unaltered expression of six proteins on chromosome 21 in fetal DS brain suggest that the existence of the trisomic state is not involved in abnormal development of fetal DS brain and that the gene dosage effect hypothesis is not sufficient to fully explain the DS phenotype. We are in the process of quantifying all gene products of chromosome 21 and our first results do not support the gene dosage hypothesis. PMID- 12624743 TI - Protein levels of genes encoded on chromosome 21 in fetal Down syndrome brain: challenging the gene dosage effect hypothesis (Part II). AB - Down syndrome (DS) is the most common genetic cause of mental retardation. To explain the impact of extra chromosome 21 in the pathology of DS, gene dosage effect hypothesis has been proposed, but several investigators including our group have challenged this hypothesis. Although analysis of the sequence of chromosome 21 has been essentially completed, the molecular and biochemical mechanisms underlying the pathology are still unknown. We therefore investigated expression levels of six proteins encoded on chromosome 21 (HACS1, DYRK1A, alphaA crystallin, FTCD, GARS-AIRS-GART, and CBS) in fetal cerebral cortex from DS and controls at 18-19 weeks of gestational age using Western blot analysis. Protein expression of HACS1 was significantly and remarkably decreased in DS, and the expression levels of five proteins were comparable between DS and controls suggesting that the gene dosage effect hypothesis is not sufficient to fully explain the DS phenotype. We are continuing to quantify proteins whose genes are encoded on chromosome 21 in order to provide a better understanding of the pathobiochemistry of DS at the protein level. PMID- 12624744 TI - Protein levels of genes encoded on chromosome 21 in fetal Down syndrome brain: challenging the gene dosage effect hypothesis (Part III). AB - Down syndrome (DS) is the most frequent genetic disorder with mental retardation and caused by trisomy 21. Although the gene dosage effect hypothesis has been proposed to explain the impact of extra chromosome 21 on the pathology of DS, a series of evidence that challenge this hypothesis has been reported. The availability of the complete sequences of genes on chromosome 21 serves now as starting point to find functional information of the gene products, but information on gene products is limited so far. We therefore evaluated expression levels of six proteins whose genes are encoded on chromosome 21 (synaptojanin-1, chromosome 21 open reading frame 2, oligomycin sensitivity confering protein, peptide 19, cystatin B and adenosine deaminase RNA-specific 2) in fetal cerebral cortex from DS and controls at 18-19 weeks of gestational age using Western blot analysis. Synaptojanin-1 and C21orf2 were increased in DS, but others were comparable between DS and controls, suggesting that the DS phenotype cannot be simply explained by gene dosage effects. We are systematically quantifying all proteins whose genes are encoded on chromosome 21 in order to provide a better understanding of the pathobiochemistry of DS at the protein level. These studies are of significance as they show for the first time protein levels that are carrying out specific function in human fetal brain with DS. PMID- 12624745 TI - Studies on the utilization of methionine sulfoxide and methionine sulfone by rumen microorganisms in vitro. AB - An in vitro experiment was conducted to test the ability of mixed rumen bacteria (B), protozoa (P), and their mixture (BP) to utilize the oxidized forms of methionine (Met) e.g., methionine sulfoxide (MSO), methionine sulfone (MSO(2)). Rumen contents were collected from fistulated goats to prepare the microbial suspensions and were anaerobically incubated at 39 degrees C for 12 h with or without MSO (1 mM) or MSO(2) (1 mM) as a substrate. Met and other related compounds produced in both the supernatants and hydrolyzates of the incubation were analyzed by HPLC. During 6- and 12-h incubation periods, MSO disappeared by 28.3 and 42.0%, 0.0 and 0.0%, and 40.6 and 62.4% in B, P, and BP suspensions, respectively. Rumen bacteria and the mixture of rumen bacteria and protozoa were capable to reduce MSO to Met, and the production of Met from MSO in BP (156.6 and 196.1 micromol/g MN) was about 17.3 and 14.1% higher than that in B alone (133.5 and 171.9 micromol/g MN) during 6- and 12-h incubations, respectively. On the other hand, mixed rumen protozoa were unable to utilize MSO. Other metabolites produced from MSO were found to be MSO(2) and 2-aminobutyric acid (2AB) in B and BP. MSO(2) as a substrate remained without diminution in all-microbial suspensions. It was concluded that B, P, and BP cannot utilize MSO(2); but MSO can be utilized by B and BP for producing Met. PMID- 12624746 TI - Preservation of amino acids during long term ischemia and subsequent reflow with supplementation of L-arginine, the nitric oxide precursor, in the rat heart. AB - We investigated whether L-arginine, used in heart preservation to limit endothelial damage, may influence the pool of amino acids during long term ischemia and reflow. Isolated isovolumic rat hearts (n = 23) were submitted to 8 h of hypothermic ischemia after cardioplegic arrest with the Centre de Resonance Magnetique Biologique et Medicale (CRMBM) solution with or without L-arginine (Arg and No Arg groups respectively). Hearts were freeze-clamped after ischemia (n = 11) or submitted to 60 min of reflow (n = 12) and freeze-clamped. Eight hearts were perfused aerobically for 20 min and freeze-clamped (No ischemia group). Addition of L-arginine to the CRMBM solution limited aspartate depletion and decreased lysine level at the end of ischemia. After reflow, L-arginine supplementation increased the pool of glutamate and arginine and limited the depletion of serine, asparagine, glycine and taurine. We conclude that adding L arginine to the CRMBM cardioplegic solution during long term ischemia preserved the amino acids pool. PMID- 12624747 TI - Determination of the standard Gibbs energies of transfer of cations and anions of amino acids and small peptides across the water nitrobenzene interface. AB - The standard Gibbs energies of transfer of anions and cations of amino acids and small peptides across the water nitrobenzene interface were determined with the help of a novel electrochemical technique using three-phase electrodes. This is the first time that reliable data are reported for the anions of amino acids. The main result is that the standard Gibbs energies of transfer of the anion and cation of an amino acid are almost the same. PMID- 12624748 TI - Taurine-evoked chloride current and its potentiation by intracellular Ca2+ in immature rat hippocampal CA1 neurons. AB - Taurine is one of the most abundant free amino acids in the immature mammalian central nervous system. In the present study, whole-cell patch-clamp recordings were made to examine taurine-evoked currents ( I(Tau)) in acutely dissociated immature rat hippocampal CA1 neurons. Taurine at low concentrations (/=3 mM) activated both glycine and GABA(A) receptors. Moreover, elevation of intracellular Ca(2+) via non-NMDA receptor activation enhanced I(Tau) reversibly. The results indicate that taurine may act as a native ligand of glycine receptors and modulate neurotransmissions in the immature hippocampus, and under certain conditions it can also activate GABA(A) receptors. The potentiation of I(Tau) by intracellular Ca(2+) may contribute to the protection effect of taurine under some cell damaging conditions. PMID- 12624749 TI - Effect of D-amino acids on some mitochondrial functions in rat liver. AB - We studied the role of the D-amino acids (D-aa) D-serine, D-alanine, D methionine, D-aspartate, D-tyrosine and D-arginine on rat liver mitochondria. The stability of D-amino acids, mitochondrial swelling, transmembrane potential and oxygen consumption were studied under oxidative stress conditions in rat liver mitochondria. In the presence of glutamate-malate all D-aas salts increased mitochondrial swelling, while in the presence of succinate plus rotenone only D ala, D-arg and D-ser, induced mitochondrial swelling. The transmembrane potential (deltapsi) was decreased in the presence of 1 microM Ca(2+). The D-aas inhibited oxygen consumption in state 3. The D-aa studied exerted effects on mitochondria via an increase of free radicals production. PMID- 12624750 TI - Volatile constituents of glutathione--ribose model system and its antioxidant activity. AB - Reaction between glutathione and ribose was carried out to study the volatiles formed via Maillard reaction and their antioxidant activity as well as their role in inhibition of LDL oxidation. The simultaneous distillation-extraction technique was used for trapping the volatile components followed by GC-MS analysis. Thirty six compounds were identified with the predominance of carbonyls and sulfur-containing compounds in the volatiles of this model system. Sensory evaluation was performed for the model system product according to the International Standard Methods (ISO). The results showed a high decrease in roasted and burnt attributes and remarkable increase in the like-boiled and roasted meat attributes. The sensory results of the model system product were confirmed by the presence of high concentrations of some volatile compounds having meat--like aroma such as 2-methyl-3-furanthiol and 2-furylmethanethiol. The radical scavenging activity of glutathione-ribose model system was quantified spectrophotometrically, using DPPH radical. The activity of the model system product was found to be slightly lower than that of gallic acid and BHA, but it was much higher than that of cinnamic acid (200 ppm. for each). A highly antioxidative activity was recorded by the model system product during the inhibition of LDL-oxidation in comparison with L-ascorbic acid as well as reduced glutathione (as a concentration of 0.5 micromol/L, for each) which may be due to the presence of some compounds such as 2-furylmethanethiol, 2-acetyl thiazole, 4 hydroxy-5-methyl-3(2H)-furanone. PMID- 12624751 TI - Isolation of the bifunctional enzyme lysine 2-oxoglutarate reductase-saccharopine dehydrogenase from Phaseolus vulgaris. AB - Lysine is catabolyzed by the bifunctional enzyme lysine 2-oxoglutarate reductase saccharopine dehydrogenase (LOR-SDH) in both animals and plants. LOR condenses lysine and 2-oxoglutarate into saccharopine, using NADPH as cofactor and SDH converts saccharopine into alpha-aminoadipate delta-semialdehyde and glutamic acid, using NAD as cofactor. The distribution pattern of LOR and SDH among different tissues of Phaseolus vulgaris was determined. The hypocotyl contained the highest specific activity, whereas in seeds the activities of LOR and SDH were below the limit of detection. Precipitation of hypocotyl proteins with increasing concentrations of PEG 8000 revealed one broad peak of SDH activity, indicating that two isoforms may be present, a bifunctional LOR-SDH and possibly a monofunctional SDH. During the purification of the hypocotyl enzyme, the LOR activity proved to be very unstable, following ion-exchange chromatography. Depending on the purification procedure, the protein eluted as a monomer of 91-94 kDa containing only SDH activity, or as a dimer of 190 kDa with both, LOR and SDH activities, eluting together. PMID- 12624752 TI - Polyamine oxidase activity in rats treated with mitoguazone: specific and permanent decrease in thymus. AB - To extend the knowledge on the role of polyamine oxidase in thymus physiology, we evaluated the in vivo effect of the polyamine biosynthetic pathway inhibitor mitoguazone. The drug markedly and permanently decreased the enzyme activity in the organ, in which the level of putrescine also decreased at the later times observed. A byproduct of the reaction catalyzed by polyamine oxidase is hydrogen peroxide, a well known inducer of apoptosis. The decrease in polyamine oxidase activity, with the consequent decrease in hydrogen peroxide production, is correlated with a positive effect on thymus physiology. Since mitoguazone has been successfully employed in patients with AIDS-related diseases, in which the reconstitution of the immune function is a favorable prognostic index, we hypothesized that mitoguazone may have the thymus as target organ, and that the decrease in polyamine oxidase activity may have a role in the positive effect of the drug. PMID- 12624753 TI - Production of cysteine for bacterial and plant biotechnology: application of cysteine feedback-insensitive isoforms of serine acetyltransferase. AB - The first step of cysteine biosynthesis in bacteria and plants consists in the formation of O-acetylserine catalyzed by serine acetyltransferase (SAT). SAT is highly sensitive to feedback inhibition by cysteine as part of the regulatory circuit of cysteine biosynthesis und thus hampers over-expression and fermentation of cysteine in biotechnological production processes. Since plants contain multiple SAT isoforms with different cysteine feedback sensitivity, this resource was exploited to demonstrate the suitability of plant SATs for the production of cysteine in both bacteria and plants. Three new cDNAs encoding SATs were isolated from Nicotiana tabacum. The catalytic activity of SAT4 was insensitive up to 0.6 mM cysteine. Expression of SAT4 in a newly constructed Escherichia coli host strain without endogenous SAT activity yielded a significant accumulation of cysteine in the culture medium compared to expression of cysteine sensitive SATs in the same strain. The application of a similarly insensitive SAT isoform from A. thaliana demonstrated the suitability of this approach to increase cysteine levels in transgenic tobacco plants. PMID- 12624754 TI - Free amino acids in the haemolymph of honey bee queens (Apis mellifera L.). AB - In queen honey bees the free amino acid content in the haemolymph clearly depends on the physiological function and social environment of the individual. While in drones and workers the content of free amino acids increases after emergence until it reaches a peak in 5-day-old animals and decreases afterwards, the amino acid content in queens reaches its highest level (>60 nmol/ microl haemolymph) with the onset of egg laying (10 d of age). This level is about 2.5 times more than the highest level found in workers. Queens maintain this high level also when they are older (>30 d) and continue to lay eggs in average colonies. As in drones and workers, in queens the predominant amino acid is proline, which accounts for more than 50% of the total content of free amino acids in egg-laying individuals. When 10-day-old queens are prevented from mating and do not lay eggs, their amino acid content is significantly lower compared to laying queens of the same age. Also the social environment influences the contents of free amino acids in queens. When virgin queens were kept for 6 days with 20 worker bees and sufficient honey and pollen in an incubator, they had significantly lower concentrations of amino acids than virgin queens living for the same period with about 8000 workers in a colony. Most probably, the high amino acid concentration in the haemolymph is the basis for the high protein synthesis activity of laying queens. PMID- 12624755 TI - Characterization of N-methyl-D-aspartate-evoked taurine release in the developing and adult mouse hippocampus. AB - Taurine is an inhibitory amino acid acting as an osmoregulator and neuroromodulator in the brain, with neuroprotective properties. The ionotropic glutamate receptor agonist N-methyl-D-aspartate (NMDA) greatly potentiates taurine release from brain preparations in both normal and ischemic conditions, the effect being particularly marked in the developing hippocampus. We now characterized the regulation of NMDA-stimulated taurine release from hippocampal slices from adult (3-month-old) and developing (7-day-old) mouse using a superfusion system. The NMDA-stimulated taurine release was receptor-mediated in both adult and developing mouse hippocampus. In adults, only NO-generating compounds, sodium nitroprusside, S-nitroso-N-acetylpenicillamine and hydroxylamine reduced the release, as did also NO synthase inhibitors, 7 nitroindazole and nitroarginine, indicating that the release is mediated by the NO/cGMP pathway. On the other hand, the regulation of the NMDA-evoked taurine release proved to be somewhat complex in the immature hippocampus. It was not affected by the NOergic compounds, but enhanced by the protein kinase C activator 4 beta-phorbol 12-myristate 13-acetate and adenosine receptor A(1) agonists, N(6) cyclohexyladenosine and R(-)N(6)-(2-phenylisopropyl)adenosine in a receptor mediated manner. The activation of both ionotropic 2-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) receptors and metabotropic glutamate group I receptors also enhanced the evoked release. The NMDA-receptor-stimulated taurine release could be a part of the neuroprotective properties of taurine, being important particularly under cell-damaging conditions in the developing hippocampus and hence preventing excitotoxicity. PMID- 12624756 TI - Hamster D-amino-acid oxidase cDNA: rodents lack the 27th amino acid residue in D amino-acid oxidase. AB - The nucleotide sequence of cDNA that encodes hamster d-amino-acid oxidase (DAO) was determined. The cDNA consisted of 1,590 nucleotides and a poly(A) tail. It had an open reading frame for a protein consisting of 346 amino acid residues. The number of the amino acid residues is the same as that of the rat DAO. However, the hamster DAO has one residue more than mouse DAO and one residue less than human, pig, rabbit, and guinea pig DAOs. Amino acid sequence of the hamster DAO was highly similar to those of mouse and rat DAOs: 89% and 88% of the amino acid residues were identical between the hamster and mouse DAOs and between the hamster and rat DAOs, respectively. The homology was slightly less between the hamster DAO and the human (81%), pig (78%), rabbit (78%), or guinea pig DAO (82%). It has been proposed that the mouse and rat DAOs lack an amino acid residue corresponding to the 25th residue of the DAOs of other mammals. However, a detailed comparison of the amino acid sequences as well as the underlying nucleotide sequences by inclusion of the hamster ones revealed that the rodent DAOs does not lack the 25th, but the 27th residue. PMID- 12624758 TI - Genetic analysis of a complex trait in the Utah Genetic Reference Project: a major locus for PTC taste ability on chromosome 7q and a secondary locus on chromosome 16p. AB - The ability to taste phenylthiocarbamide (PTC) shows complex inheritance in humans. We obtained a quantitative measure of PTC tasting ability in 267 members of 26 large three-generation families that were part of a set of CEPH families that had been used for genetic mapping. Significant bimodality was found for the distribution of age and gender adjusted scores (P<0.001), with estimated means of 3.16 (SD=1.80) and 9.26 (SD=1.54). Using the extensive genotyping available in these families from the genetic mapping efforts, we performed a genome scan by using 1324 markers with an average spacing of 4 cM. Analyses were first carried out with a recessive genetic model that has traditionally been assumed for the trait, and a threshold score of 8.0 delineating tasters from non-tasters. In this qualitative analysis, the maximum genome-wide lod score was 4.74 at 246 cM on chromosome 7; 17 families showed segregation of the dichotomous PTC phenotype. No other lod scores were significant; the next highest score was on chromosome 10 (lod=1.64 at 85 cM), followed by chromosome 3 (lod=1.29 at 267 cM). Because PTC taste ability exhibited substantial quantitative variation, the quantitative trait was also analyzed by using a variance components approach in SOLAR. The maximum quantitative genome-wide lod score was 8.85 at 246 cM on chromosome 7. Evidence for other possible quantitative loci was found on chromosomes 1 (lod=2.31 at 344 cM) and 16 (lod=2.01 at 14 cM). A subsequent two-locus whole genome scan conditional on the chromosome 7 quantitative trait locus identified the chromosome 16 locus (two-locus lod=3.33 at 14 cM). PMID- 12624759 TI - Efficient soybean transformation using hygromycin B selection in the cotyledonary node method. AB - The efficiency of soybean [Glycine max (L.) Merrill] transformation was significantly increased from an average of 0.7% to 16.4% by combining strategies to enhance Agrobacterium tumefaciens-mediated T-DNA delivery into cotyledonary node cells with the development of a rapid, efficient selection protocol based on hygromycin B. Wounded cotyledonary-node explants were inoculated with A. tumefaciens carrying either a standard-binary or super-binary plasmid and co cultivated in the presence of mixtures of the thiol compounds, L-cysteine, dithiothreitol, and sodium thiosulfate. Transformed shoots began elongating only 8 weeks after co-cultivation. Southern analysis confirmed integration of the T DNA into genomic DNA and revealed no correlation between the complexity of the integration pattern and thiol treatment applied at co-cultivation. All T(0) plants were fertile and the majority of the lines transmitted the beta glucuronidase (GUS) phenotype in 3:1 or 15:1 ratios to their progenies. PMID- 12624760 TI - Two isoforms of Rubisco activase in cotton, the products of separate genes not alternative splicing. AB - In several plant species, ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activase consists of two isoforms that are produced by alternative splicing of a pre-mRNA. Two forms of activase corresponding to the longer, redox regulated alpha and the shorter, beta forms were detected immunologically in cotton (Gossypium hirsutum L.) leaves, but their N-termini differed in 4 of 14 residues. The cDNAs for the alpha and beta forms of cotton activase diverged throughout the translated and 3'-untranslated regions, including variations that accounted for the differences in N-terminal amino acid sequence. Analysis of genomic DNA confirmed that separate genes encoded the alpha and beta forms of cotton activase. Separate activase genes were also detected in diploid species of cotton containing the different progenitor genomes of the cultivated allotetraploid, indicating that the occurrence of separate alpha- and beta-form genes in cotton predates the merger of the diploid genomes. The deduced amino acid sequences of the two forms of cotton activase exhibited 84% identity and both forms were active after expression in Escherichia coli. The recombinant alpha and beta forms exhibited similar affinities for ATP and only minor differences in thermotolerance, but their ATPase specific activities differed. The results show for the first time a plant species with two forms of activase that are structurally and functionally equivalent to the alternatively spliced alpha and beta forms in other plants, but that are encoded by separate genes. That cotton still expresses both forms of activase, even without alternative splicing, suggests that each form has a required function in photosynthesis. PMID- 12624762 TI - Beta-ketoacyl-acyl carrier protein synthase III from pea (Pisum sativum L.): properties, inhibition by a novel thiolactomycin analogue and isolation of a cDNA clone encoding the enzyme. AB - A beta-ketoacyl-acyl carrier protein (ACP) synthase III (KAS III; short-chain condensing enzyme) has been partly purified from pea leaves. The enzyme, which had acetyl-CoA:ACP acyltransferase (ACAT) activity, was resolved from a second, specific, ACAT protein. The KAS III enzyme had a derived molecular mass of 42 kDa (from its cDNA sequence) and operated as a dimer. Its enzymological characteristics were similar to those of two other plant KAS III enzymes except for its inhibition by thiolactomycin. A derivative of thiolactomycin containing a longer (C8 saturated) hydrophobic side-chain (compound 332) was a more effective inhibitor of pea KAS III and showed competitive inhibition towards malonyl-ACP whereas thiolactomycin showed uncompetitive characteristics at high concentrations. This difference may be due to the better fit of compound 332 into a hydrophobic pocket at the active site. A full-length cDNA for the pea KAS III was isolated. This was expressed in Escherichia coli as a fusion protein with glutathione S-transferase in order to facilitate subsequent purification. Demonstrated activity in preparations from E. coli confirmed that the cDNA encoded a KAS III enzyme. Furthermore, the expressed KAS III had ACAT activity, showing that the latter was inherent. The derived amino acid sequence of the pea cDNA showed 81-87% similarity to that for other plant dicotyledon KAS IIIs, somewhat less for Allium porrum (leek, 71%) and for Porphyra spp. (62%), Synechocystis spp. (65%) and various bacteria (42-65%). The pea KAS III exhibited four areas of homology, three of which were around the active-site Cys(123), His(323) and Asn(353). In addition, a stretch of 23 amino acids (residues 207-229 in the pea KAS III) was almost completely conserved in the plant KAS IIIs. Modelling this stretch showed they belonged to a peptide fragment that fitted over the active site and contained segments suggested to be involved in substrate binding and in conformational changes during catalysis, as well as an arginine suggested to participate in the acid-base catalytic mechanism. PMID- 12624761 TI - Functional identification of AtTPS03 as (E)-beta-ocimene synthase: a monoterpene synthase catalyzing jasmonate- and wound-induced volatile formation in Arabidopsis thaliana. AB - (E)-beta-Ocimene is one of the most commonly found monoterpenes of the volatile blends that are emitted from leaves in response to damage by herbivores or mechanical wounding. (E)-beta-Ocimene is also a component of many floral scents. Airborne (E)-beta-ocimene emitted from plants can serve as a chemical cue for the attraction of parasitoids or predators of plant herbivores and also as an attractant for pollinating insects. Furthermore, exposure of plants to (E)-beta ocimene can activate defense gene expression. In this paper, we describe cDNA cloning and functional characterization of a gene encoding a highly specialized (E)-beta-ocimene synthase, AtTPS03, from Arabidopsis thaliana (L.) Heynh. AtTPS03 was identified as a member of the large AtTPS gene family of putative terpene synthases. A cDNA for AtTPS03 was expressed in Escherichia coli and the enzyme function identified in vitro. The A. thaliana (E)-beta-ocimene synthase produces almost exclusively (E)-beta-ocimene (94%) with minor amounts of the related acyclic monoterpenes (Z)-beta-ocimene (4%) and myrcene (2%). Transcripts for AtTPS03 were up-regulated in leaves of Arabidopsis in response to mechanical wounding and treatment with jasmonic acid, concurrent with induced emission of (E)-beta-ocimene. AtTPS03 provides an important gene for probing plant-insect and possibly plant-plant interactions mediated by terpenoid volatiles. PMID- 12624763 TI - Proteomic identification of plant proteins probed by mammalian nitric oxide synthase antibodies. AB - Several studies suggest that a mammalian-like nitric oxide synthase (NOS) exists in plants. Researchers have attempted to verify its presence using two approaches: (i) determination of NOS functional activity and (ii) probing with mammalian NOS antibodies. However, up to now, neither a NOS-like gene nor a protein has been found in plants. While there is still some controversy over whether the NOS functional activity seen is due to nitrate reductase, using the mammalian NOS antibodies in western blot analysis, several groups have reported the presence of immunoreactive protein bands in plant homogenates. Based on these results, immunohistochemical studies using these antibodies have also been used to localize NOS in plant tissues. However, plant NOS has never been positively identified or characterized. Thus, we used a proteomic approach to verify the identities of plant proteins that cross-reacted with the mammalian NOS antibodies. Proteins extracted from maize (Zea mays L.) embryonic axes were separated by two-dimensional gel electrophoresis and subjected to western blot analysis with the mammalian neuronal NOS and inducible NOS antibodies. Twenty immunoreactive protein spots recognized on a corresponding Coomassie blue-stained two-dimensional gel were subjected to tryptic digestion, followed by identification using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Fifteen proteins were successfully identified and they have described functions that are unrelated to NO metabolism. The remaining five proteins could not be identified. The amino acid sequences of these identified proteins and those used to raise the antibodies were aligned. However, no homologous region could be found. Our results demonstrate that the mammalian NOS antibodies recognize many NOS-unrelated plant proteins. Therefore, it is inappropriate to infer the presence of plant NOS using this immunological technique. PMID- 12624764 TI - Accumulation response of chloroplasts induced by mechanical stimulation in bryophyte cells. AB - Chloroplast movement has been studied in many plants but mainly as a model system for light signaling. However, we recently showed that the avoidance response of chloroplasts is also induced by mechanical stimulation in fern protonemal cells. Here we report the discovery of a mechanically induced accumulation response of chloroplasts in bryophytes. When mechanical stimulation was directly applied with a capillary to a part of a cell, chloroplasts moved towards and accumulated at the pressed site within 30 min after the onset of stimulation in all species tested. The accumulation movement of chloroplasts was inhibited by Cremart but not by cytochalasin B in red-light-grown protonemata of Physcomitrella patens (Hedw.) B., S. & G. To determine the contribution of external Ca(2+) to the response, we examined the effects on the accumulation movement of gadolinium (Ga(3+)), an inhibitor of stretch-activated ion channels, and lanthanum (La(3+)), a potent inhibitor of calcium channels. Mechano-relocation of chloroplasts was abolished by these drugs, but no effects were observed on photo-relocation of chloroplasts, irrespective of light colors and intensity. These results suggest that influx of external Ca(2+) through the plasma membrane is essential for the early steps in signaling of mechano-relocation of chloroplasts whose motility system is dependent on microtubules. PMID- 12624765 TI - The transformation of anthers in the msca1 mutant of maize. AB - In normal anther development in maize (Zea mays L), large hypodermal cells in anther primordia undergo a series of proscribed cell divisions to form an anther containing microsporogenous cells and three distinctive anther wall layers: the tapetum, the middle layer and the endothecium. In homozygous msca1 mutants of maize, stamen primordia are initiated normally and large hypodermal cells can be detected in developing anthers. However, the normal series of cell division and differentiation events does not occur in msca1 mutant plants. Rather, structures containing parenchymal cells and ectopic, nonfunctional vascular strands are formed. The epidermal surfaces of these structures contain stomata, which are normally absent in maize anthers. Thus, all of the cell layers of the "anther" have been transformed in mutant plants. The filaments of the mutant structures are normal, and all other flower parts are normal. The msca1 mutation does not affect female fertility, but transformed "stamen" structures remain associated with mature ovules rather than aborting as in normal ear development. The msca1 mutation is distinctive in that only one part of a single (male) reproductive organ is transformed. The resulting structure has general vegetative features, but cannot be conclusively identified as a particular vegetative organ. PMID- 12624766 TI - Integrating diel starch metabolism with the circadian and environmental regulation of Crassulacean acid metabolism in Mesembryanthemum crystallinum. AB - The diel (24-h) Crassulacean acid metabolism (CAM) cycle in Mesembryanthemum crystallinum (L.) requires rhythmic patterns of transitory starch degradation to produce carbon skeletons for phospho enolpyruvate (PEP) synthesis during the nocturnal Phase I, when PEP carboxylase (PEPc) mediates CO(2) fixation. Under a normal light-dark cycle, nocturnal malate accumulation and nocturnal CO(2) uptake were observed for CAM-induced, but not C(3), M. crystallinum. In both C(3) and CAM plants, transcripts encoding beta-amylase and starch phosphorylase accumulated during the afternoon and declined nocturnally. Under a continuous light regime, ribulose-1,5-bisphosphate carboxylase/oxygenase activity remained co-ordinated with the CAM phases, and circadian abundance patterns were observed for transcripts encoding starch degradative enzymes. Despite circadian PEPc kinase expression, the accumulation of vacuolar malate ceased under continuous light. Exposure to CO(2)-free air for 24 h inhibited starch accumulation over the photoperiod, but re-fixation of respiratory CO(2) resulted in the overnight accumulation of malate to levels comparable to those of control plants. Upon return to normal air, the depleted starch concentration led to stoichiometric decreases in Phase-I CO(2) uptake and malate accumulation. The up-regulation of PEPc kinase transcripts under these conditions was ineffective at sustaining Phase-I CO(2) uptake under starch-limited conditions. We conclude that starch turnover regulates and limits carbon flux through the diel CAM cycle. PMID- 12624767 TI - Determination of the starch-phosphorylating enzyme activity in plant extracts. AB - For quantification of alpha-glucan, water dikinase (GWD) activity in crude extracts of plant tissues a radio-labeling assay was established that uses soluble starch and (33)P-labeled ATP as phosphate acceptor and donor, respectively. A constant rate of starch labeling was observed only if the ATP applied was labeled at the beta position. In wild-type extracts from leaves of Arabidopsis thaliana (L.) Heynh. the maximum rate of starch phosphorylation was approximately 27 pmol min(-1) (mg protein)(-1). Leaf extracts from the GWD deficient sex1 mutants of Arabidopsis showed no significant incorporation of phosphate whereas extracts from potato (Solanum tuberosum L.) tuber expressing a GWD antisense construct exhibited less activity than the wild-type control. To our knowledge this is the first time that a quantification of the starch phosphorylating activity has been achieved in plant crude extracts. PMID- 12624768 TI - Activation of three pathogen-inducible promoters of tobacco in transgenic pear (Pyrus communis L.) after abiotic and biotic elicitation. AB - In order to improve pear resistance against fire blight caused by Erwinia amylovora, a search for promoters driving high-level expression of transgenes specifically in response to this bacterial pathogen has been undertaken. We have examined the ability of hsr203J, str246C and sgd24 tobacco (Nicotiana tabacum L.) promoters to drive expression of the uidA reporter gene in pear. Transgenic pear clones were obtained by Agrobacterium tumefaciens-mediated transformation. Beta glucuronidase activity was determined quantitatively and qualitatively in these plants grown in vitro using fluorometric and histochemical assays and compared to cauliflower mosaic virus (CaMV) 35S promoter-driven activity. The hsr203J promoter appeared to be very weakly activated following inoculation in pear, which is the converse of the situation in tobacco. The str246C promoter was rapidly activated in pear during compatible and incompatible interactions, by wounding and following the application of several elicitors (capsicein, cryptogein, harpin, salicylic acid and jasmonic acid). The sgd24 promoter, a deletion derivative of str246C, exhibited a low level of expression after bacterial inoculation, was weakly activated by wounding and elicitors, and was not activated by phytohormones (salicylic acid and jasmonic acid). Interestingly, the sgd24 promoter was locally activated in pear, whereas the str246C promoter was activated systemically from the infection site. Taken together, these data show that, although the s tr246C and sgd24 promoters are less active than the CaMV35S promoter in pear, their pathogen-responsiveness would permit them to be used to drive the expression of transgenes to promote bacterial disease resistance. PMID- 12624769 TI - Plastid transcription in the holoparasitic plant genus Cuscuta: parallel loss of the rrn16 PEP-promoter and of the rpoA and rpoB genes coding for the plastid encoded RNA polymerase. AB - The holoparasitic plant genus Cuscuta comprises a range of species whose plastid genomes have different degrees of reductions in their coding capacity. In this study, four Cuscuta species, Cuscuta reflexa, C. gronovii, C. subinclusa and C. odorata, that possess substantial physiological differences, were analysed with respect to the sequence and promoter structure of the rrn16 gene coding for the ribosomal 16S rRNA. Whereas the coding region of this gene is highly conserved among all four Cuscuta species, significant differences were observed in the non coding region 5' of rrn16 with respect to both the length of the intergenic region between rrn16 and trnV and the promoters used to initiate transcription of the rrn16 gene. In the green species C. reflexa, rrn16 transcription starts from a functional plastid-encoded RNA polymerase (PEP) promoter that is missing in the other three species, C. gronovii, C. odorata and C. subinclusa. Instead, a 15 nucleotide-long conserved sequence immediately upstream of the mapped 5' ends bearing the nuclear-encoded RNA polymerase (NEP) promoter motif could be identified in these three species. The lack of a PEP promoter in these species coincides with the loss of two genes that encode subunits of PEP (rpoA and rpoB). PMID- 12624770 TI - The promoter of a rice glycine-rich protein gene, Osgrp-2, confers vascular specific expression in transgenic plants. AB - The genomic sequence of a rice (Oryza sativa L.) glycine-rich protein (GRP) gene, designated Osgrp-2, has been previously determined (GenBank U40708). Primer extension analysis indicated that transcription starts 47 bp upstream of the translation start codon. To gain an insight into the transcriptional regulation of this gene, the 2,401-bp promoter sequence and a series of its 5' deletions were transcriptionally fused to the beta-glucuronidase (GUS) gene. GUS activity was subsequently assayed in a transient expression system of tobacco ( Nicotiana tabacum L.) protoplasts, which revealed the presence of a positive regulatory region (-2290 to -1406) and two negative regulatory regions (-2401 to -2291 and 1405 to -1022) in the Osgrp-2 promoter for the promoter activity. The positive regulatory region displayed an enhancer-like activity when fused to the cauliflower mosaic virus (CaMV) 35S minimal promoter (-89 to +6) to drive GUS expression and assayed on tobacco leaves by the Agrobacterium-mediated transient expression technique (agroinfiltration). Histochemical staining for GUS activity on transgenic tobacco plants has further indicated a preferential expression in vascular tissues of stems and leaves conferred by the positive regulatory region. A 1,023-bp fragment of the Osgrp-2 promoter (-1021 to +2) fused with GUS was transformed into tobacco and proved to be capable of conferring vascular-specific expression. Further 5' and 3' deletion analysis of the 1,023-bp promoter revealed that a 99-bp fragment located from -497 to -399 contained cis-elements responsible for vascular-specific expression. PMID- 12624771 TI - Temperature-related non-homogeneous fatty acid desaturation in sunflower (Helianthus annuus L.) seeds. AB - The fatty acid compositions of half-seeds and whole seeds of the temperature dependent high-stearic-acid sunflower (Helianthus annuus L.) mutant CAS-14 were unexpectedly different. We found that there is a longitudinal gradient starting from the embryo up to the end of the cotyledon. The stearic acid content varied from 9.7 to 34.6% in seeds produced in a growth chamber (39/24 degrees C; day/night), and from 14.0 to 34.4% in seeds produced in the field during the summer season (35-40 degrees C in daylight and 20-25 degrees C at night). The gradient occurs throughout seed formation, and is due to a spatial and non temporal regulation of stearic acid desaturation. A similar temperature-regulated behaviour, but for oleic and linoleic acid contents, was found in normal sunflower seeds. Since the deposition of oil bodies was homogeneous during seed formation, seeds showed the gradient throughout their development. This non homogeneous distribution must be due to differences in the enzymatic pathway of de-novo fatty acid desaturation along the seed, resembling a morphogen gradient. Other high-stearic-acid mutant lines, such as CAS-3, did not show any gradient. This is the first time that a gradient and an inheritable maternal control of the fatty acid composition have been found in oilseeds. PMID- 12624773 TI - Protoxylem: the deposition of a network containing glycine-rich cell wall proteins starts in the cell corners in close association with the pectins of the middle lamella. AB - Antibodies were used to localise polysaccharide and protein networks in the protoxylem of etiolated soybean (Glycine max L.) hypocotyls. The deposition of glycine-rich proteins (GRPs) starts in the cell corners between protoxylem elements and xylem parenchyma cells. Finally, the GRPs form a network between two mature protoxylem elements. The network also interconnects the ring- and spiral shaped secondary wall thickenings, as well as the thickenings with the middle lamellae of living xylem parenchyma cells. In addition to the GRP network, a polysaccharide network composed mainly of pectins is involved in the attachment of the secondary wall thickenings to the middle lamellae of xylem parenchyma cells. PMID- 12624772 TI - Gradual shifts in sites of free-auxin production during leaf-primordium development and their role in vascular differentiation and leaf morphogenesis in Arabidopsis. AB - The major regulatory shoot signal is auxin, whose synthesis in young leaves has been a mystery. To test the leaf-venation hypothesis [R. Aloni (2001) J Plant Growth Regul 20: 22-34], the patterns of free-auxin production, movement and accumulation in developing leaf primordia of DR5::GUS-transformed Arabidopsis thaliana (L.) Heynh. were visualized. DR5::GUS expression was regarded to reflect sites of free auxin, while immunolocalization with specific monoclonal antibodies indicated total auxin distribution. The mRNA expression of key enzymes involved in the synthesis, conjugate hydrolysis, accumulation and basipetal transport of auxin, namely indole-3-glycerol-phosphate-synthase, nitrilase, IAA-amino acid hydrolase, chalcone synthase and PIN1 as an essential component of the basipetal IAA carrier, was investigated by reverse transcription-polymerase chain reaction. Near the shoot apex, stipules were the earliest sites of high free-auxin production. During early stages of primordium development, leaf apical dominance was evident from strong beta-glucuronidase activity in the elongating tip, possibly suppressing the production of free auxin in the leaf tissues below it. Hydathodes, which develop in the tip and later in the lobes, were apparently primary sites of high free-auxin production, the latter supported by auxin conjugate hydrolysis, auxin retention by the chalcone synthase-dependent action of flavonoids and also by the PIN1-component of the carrier-mediated basipetal transport. Trichomes and mesophyll cells were secondary sites of free-auxin production. During primordium development there are gradual shifts in sites and concentrations of free-auxin production occurring first in the tip of a leaf primordium, then progressing basipetally along the margins, and finally appearing also in the central regions of the lamina. This developmental pattern of free auxin production is suggested to control the basipetal maturation sequence of leaf development and vascular differentiation in Arabidopsis leaves. PMID- 12624774 TI - Increased capacity for synthesis of the D1 protein and of catalase at low temperature in leaves of cold-hardened winter rye (Secale cereale L.). AB - The effect of low temperature on protein synthesis, particularly the synthesis of the photolabile proteins D1 of photosystem II and catalase (EC 1.11.1.6), was compared in non-hardened leaves (NHL) and cold-hardened leaves (CHL) of winter rye (Secale cereale L.). At 4 degrees C, both the uptake of L-[(35)S]methionine into leaf sections and its incorporation into proteins were reduced, relative to 25 degrees C. However, much lower reductions were observed in CHL than in NHL. In particular, the proportion of the L-[(35)S]methionine uptake incorporated into membrane proteins at 4 degrees C was considerably higher in CHL than in NHL. At 25 degrees C, the incorporation of L-[(35)S]methionine into both the D1 protein and catalase was lower in CHL than in NHL, in accord with a slower light-induced turnover in CHL. At 4 degrees C, the incorporation into the D1 protein and catalase was, however, much higher in CHL than in NHL, indicating that their de novo synthesis was less suppressed by the low temperature. The results indicate that cold-acclimated leaves had an improved ability to repair the photolabile proteins D1 and catalase at low temperature, relative to NHL. mRNAs for the D1 protein and for leaf catalase were not increased in CHL, relative to NHL. The superior capacity of CHL for repair at low temperature must result from posttranscriptional mechanisms. The translational efficiency of the catalase mRNA was similarly increased in both NHL and CHL during 7-h exposures to high light at 4 degrees C, while the amounts of the catalase transcript declined under these conditions. However, during a recovery period at 22 degrees C, subsequent to an exposure of NHL to 4 degrees C and high light, transient increases of the D1 and catalase mRNAs were observed. PMID- 12624775 TI - Isolation and characterization of a myo-inositol 1-phosphate synthase cDNA from developing sesame (Sesamum indicum L.) seeds: functional and differential expression, and salt-induced transcription during germination. AB - A cDNA (SeMIPS1) encoding myo-inositol 1-phosphate synthase (EC 5.5.1.4) (MIPS) has been characterized from sesame (Sesamum indicum L. cv. Dan-Baek) seeds and its functional expression analyzed. The SeMIPS1 protein was highly homologous with those from other plant species (88-94%), while a much lower degree of sequence homology (53-62%) was found with other organisms such as humans, mouse, algae, yeast, Drosophila, bacteria and other prokaryotes. A yeast-based complementation assay in yeast mutants containing a disrupted INO1gene for yeast MIPS confirmed that the SeMIPS1 gene encodes a functional MIPS. Phylogenetic analysis suggested that the SeMIPS1 gene diverged as a different subfamily or family member. Southern hybridization revealed several copies of the SeMIPS1 gene present in the sesame genome and northern blotting indicated that expression of the SeMIPS1gene may be organ specific. Salt stress during sesame seed germination had an adverse influence on transcription of SeMIPS1and greatly reduced transcript levels as the duration of exposure to a saline environment increased and NaCl concentration increased. Germination initiation of sesame seeds was severely delayed as NaCl level increased. These results suggest that expression of SeMIPS1 is down-regulated by salt stress during sesame seed germination. PMID- 12624776 TI - Histochemical analysis of phenylphenalenone-related compounds in Xiphidium caeruleum (Haemodoraceae). AB - Phenylphenalenones represent a typical group of secondary metabolites of the Haemodoraceae. Some of these phenolic compounds show organ-specific distribution within the plant. However, detailed information on cellular localisation is still lacking. To this end, confocal laser-scanning microscopy, microspectral photometry and high-performance liquid chromatography were used to study the tissue localisation of phenylphenalenone-type compounds in Xiphidium caeruleum Aubl. From the autofluorescence potential of these compounds, specific distribution of allophanylglucosides and non-glucosidic compounds of the phenylphenalenone-type in distinct cells of the roots (apical meristem, cortex, cap, epidermis) and the shoot system was revealed. Fluorescence enhancement using "Naturstoff reagent A" (NA) indicated the occurrence of NA-positive natural products in the vacuoles of leaf epidermal cells. The present results provide new insights into the possible functions of phenylphenalenone-related compounds in the context of their localisation. Additionally, the advantages and limitations of the techniques are discussed. PMID- 12624778 TI - Number of Langerhans immune cells in painful and non-painful human skin after shingles. AB - During injury or inflammation, paracrine sensitization of peripheral sensory neurons by immune cells contributes to the sensation of pain. It is less clear whether this neural sensitization contributes to neuropathic pain after neural injury as well. Shingles (herpes zoster) is a common disease that leaves some patients with prolonged neuropathic pain known as postherpetic neuralgia (PHN). Sensitization of cutaneous neurons has been hypothesized to contribute to PHN. Langerhans cells (LC), the Ia(+) macrophages of the skin, contact epidermal neurites and, when activated, synthesize molecules with the ability to sensitize axons. For these reasons, we examined morphological evidence for activation of LC in subjects with established PHN. We also evaluated the relationship between numbers of LC and nociceptive epidermal nerve endings; these are markedly reduced in PHN. We used design-based stereology to estimate the number of CD1a(+) LC in biopsies of painful and non-painful skin from ten adults with or without PHN after shingles on the torso. There were no differences in the number of LC in previously shingles-affected and normal-control skin biopsies. The number of LC also remained at normal levels in biopsies with near-loss of innervation from shingles. LC numbers were unrelated to the presence or severity of pain. These data suggest that neuropathic pain in established PHN is not associated with increased numbers of cutaneous macrophages, and that the number of cutaneous macrophages in skin from the human torso is independent of the number of epidermal nerve endings. PMID- 12624779 TI - Strong expression of CD40, CD54 and HLA-DR antigen and lack of evidence for direct cellular cytotoxicity are unique immunohistopathological features in alopecia areata. AB - The pathological role played by T cells infiltrating hair follicles in lesions of alopecia areata (AA) is unknown. We examined the expression in cryostat sections of scalp skin obtained from a total of 28 patients with AA and from five normal control subjects of (1) molecules related to the induction of cell death including Fas, Fas ligand (FasL), perforin, granzyme B (GB), and TIA-1, (2) molecules related to antigen presentation including CD1a, CD40, CD54, CD80, and CD86, and (3) molecules induced by interferon gamma (IFN-gamma) including CD40, CD54, Fas, and HLA-DR. CD3(+) T cells infiltrated perifollicularly, perivascularly and in the hair structure and there was a predominance of CD4(+) over CD8(+) cells. Antigen-presenting cells expressing CD1a, CD40, CD54, or HLA DR were also seen. Expression of CD40, CD54, HLA-DR and CD95 was also seen in the hair structure including the dermal papilla. Consistent with these observations, IFN-gamma-producing cells were also detected in the perifollicular infiltrate. In contrast, few Fas-L(+), perforin(+), GB(+) or TIA-1(+) cells were found adjacent to the follicles. Apoptotic cells were recognized only in the outer root sheath of catagen hairs. These findings suggest that infiltrating T cells interact with perifollicular or follicular antigen-presenting cells to produce IFN-gamma, which deprives dermal papilla cells of their ability to maintain anagen hair growth. PMID- 12624780 TI - Effects of metronidazole and tinidazole ointments on models for inflammatory dermatitis in mice. AB - We investigated the effects of 1-4% ointments of metronidazole and tinidazole (derivatives of nitroimidazole) on models of inflammatory dermatitis evoked by antigen, hapten and monoclonal anti-dinitrophenol (DNP) IgE antibody in mice. Metronidazole and tinidazole ointments (1) suppressed the late-phase reaction (LPR) of biphasic ear edema in mice sensitized with ovalbumin (OA), (2) suppressed trinitrochlorobenzene-induced inflammatory dermatitis, (3) suppressed the immediate phase reactions and LPR in mice passively sensitized with anti-DNP IgE mAb, and (4) enhanced vascular permeability and the number of scratching reactions, presumably due to itching, in passively sensitized mice. These results strongly indicate that metronidazole and tinidazole 1-4% ointments possess antiinflammatory, immunosuppressive and anti-itching effects, and have the potential for clinical use in the treatment of human inflammatory skin diseases including atopic dermatitis in addition to rosacea and acne vulgaris. PMID- 12624781 TI - Activation of p38 MAPK suppresses matrix metalloproteinase-1 gene expression induced by platelet-derived growth factor. AB - p38 mitogen-activated protein kinase (MAPK) regulates matrix metalloproteinase-1 (MMP-1) gene expression bidirectionally depending on the induction. We sought to determine whether cytokines related to the regulation of extracellular matrix could activate p38 MAPK in dermal fibroblasts. We determined p38 MAPK phosphorylation/activation in dermal fibroblasts stimulated with platelet-derived growth factor-BB (PDGF-BB), transforming growth factor-beta or interleukin-4. Induction of MMP-1 mRNA by PDGF-BB was enhanced in the presence of a specific inhibitor of p38 MAPK, suggesting that p38 MAPK would function as a negative regulator of the MMP-1 mRNA level. We then determined which isoforms of p38 MAPK expressed in dermal fibroblasts were responsible for the downregulation of the MMP-1 mRNA level. Overexpression of p38beta2, but not of p38alpha, significantly decreased PDGF-BB-induced MMP-1 promoter activity, although PDGF-BB activated signaling pathways to both p38alpha and p38beta2. Taken together, the results of this study indicate that p38beta2 can function as a negative regulator of MMP-1 induced by PDGF-BB in vitro, suggesting that activation of p38beta2 might contribute to the pathogenesis of cutaneous fibrosis. PMID- 12624782 TI - Cyclosporin A and methotrexate are equally effective in reducing T cell numbers in psoriatic skin lesions but have no consistent effect on IFN-gamma and IL-4 expression in psoriatic skin in situ. PMID- 12624783 TI - Expression of CD134 and CD134 ligand in lesional and nonlesional psoriatic skin. PMID- 12624784 TI - Extraneural metastases of paediatric brain tumours. AB - A survey of published cases of extraneural metastases of primary brain tumours in children under the age of 18 years revealed 245 cases; 60.7% occurred in boys and 27.3% were directly related to the placing of a shunt. The most common entities leading to extraneural metastases were medulloblastomas (56.3%), germinomas (9.8%), glioblastomas (6.9%), ependymomas (3.7%) and pilocytic astrocytomas (2.9%). While medulloblastomas were frequent among non-shunt-related metastases (69.4%), germ cell tumours were most common among shunt-related metastases (33.3%). Metastases were encountered equally often in bone (56.3%) and visceral organs (55.5%) but were markedly rarer in lymph nodes (25.3%). Medulloblastomas showed a preference for bones (88.3% of cases), germinomas for bones (77.8%) and visceral organs (66.7%), whereas ependymomas and ependymoblastomas were more frequently found in lymph nodes and visceral organs (71.5-100%). The most frequent sites of bone metastases were the pelvis (59.3% of cases with osseous lesions), femur (54%) and vertebrae (46%). Medulloblastomas preferentially spread into pelvis, femur and vertebrae, whereas germ cell tumours mainly involved pelvis, femur and ribs and glioblastomas vertebrae and pelvis. Most frequent sites of lymph node metastases were cervical (45.8% of cases with nodular lesions) and thoracic (31.3%) with 54% of children suffering from medulloblastomas and 7.9% from germ cell tumours. The most frequent organ involved in non-shunt-related metastases were the lungs (56.5% of children with visceral lesions) followed by muscle/soft tissue (26.9%), pleura (16.4%), skin (14.9%) and liver (13.4%). Among shunt-related cases, the peritoneum was the most frequent site (86.6% of children). The mean interval between primary brain tumour and occurrence of metastasis was 22.1 months with particularly grave outcomes among medulloblastomas, non-germinomatous germ cell tumours and glioblastomas (85.5-100% deceased children), whereas 60% of children suffering from germinomas were still alive at the time of reporting. PMID- 12624785 TI - Analysis of the TP53 gene in laser-microdissected glioblastoma vasculature. AB - Malignant transformation of human gliomas is accompanied by extensive proliferation of stromal blood vessels. Recent data suggest mesenchymal transdifferentiation of neoplastic cells in various human cancers, including colon and breast cancer as well as gliosarcoma. In this study, we have analyzed proliferating stromal blood vessels in glioblastoma multiforme for the presence of mutations in the tumor suppressor gene TP53. Using tissue arrays derived from glioblastoma specimens, cases with significant immunohistochemical p53 accumulation were selected for molecular genetic detection of TP53 mutations in exons 5 to 8. None of the tumors included in this series displayed properties of gliosarcoma. Proliferating glomeruloid stromal vessels were isolated by laser microdissection from paraffin sections. In six cases, single-strand conformation polymorphism analysis for mutations of the TP53 gene in stromal blood vessels compared with adjacent tumor cells and subsequent DNA sequencing of the resulting DNA fragments were carried out. Glioblastoma cells of these cases exhibited TP53 mutations in exons 5, 7 and 8. None of these tumors showed TP53 mutations in microdissected samples from glomeruloid vessels. The absence of TP53 mutations in vascular stromal components of glioblastoma multiforme supports the hypothesis that microvascular proliferations originate from the tumor stroma and are not derived from transdifferentiated glioblastoma cells. PMID- 12624786 TI - Immunohistochemical expression of inducible nitric oxide synthase (iNOS) in human brain tumors: relationships of iNOS to superoxide dismutase (SOD) proteins (SOD1 and SOD2), Ki-67 antigen (MIB-1) and p53 protein. AB - In this study, inducible nitric oxide synthase (iNOS) expression in a series of 158 human primary brain tumors was analyzed. To gain some insight into the biological significance of iNOS expression in tumor cells, comparative immunohistochemical analyses were employed to characterize the expression of iNOS, superoxide dismutase (SOD) proteins (SOD1 and SOD2), Ki-67 antigen (MIB-1) and p53 protein in these cells. Sixteen (39.0%) of the 41 glioblastoma multiforme (GBM) specimens showed iNOS immunoreactivity. Positive immunoreactions with iNOS were also detected in 2/8 anaplastic astrocytomas, 1/17 astrocytomas, 1/14 medulloblastomas and 1/11 primitive neuroectodermal tumors, but no positive reactions were observed in oligodendrogliomas (0/11), ependymomas (0/5), schwannomas (0/21), meningiomas (0/23) or pituitary adenomas (0/7). The MIB-1 labeling index of GBMs that expressed iNOS was significantly higher than that of GBMs that did not (0.025< P <0.05, Wilcoxon rank-sum test). Unlike iNOS-negative tumors, all iNOS-positive tumors coexpressed SOD1 or SOD2. In particular, there was a significant correlation between iNOS induction and SOD1 expression (P =1.65x10(-10), Fisher's exact test) in GBM specimens. There was no significant relationship between iNOS and p53 protein in any type of primary brain tumor (P >0.05, Fisher's exact test). No significant immunohistochemical reactions with iNOS, MIB-1 or p53 protein were observed in normal brain tissue sections. We conclude that primary brain tumors express iNOS, and that iNOS expression in brain tumor cells may depend, in part, on cellular proliferation potential. Based on the fact that SOD1 scavenges oxidative-stress species originating from large amounts of nitric oxide (NO) produced by iNOS, iNOS-expressing brain tumor cells may protect themselves against NO cytotoxicity by overinducing SOD1. PMID- 12624787 TI - Clinical and neuropathological correlates of Lewy body disease. AB - We investigated distribution of neuronal and glial inclusions in 30 brains obtained at autopsy from patients with Lewy bodies (LBs) disease, which was clinically diagnosed as Parkinson's disease (PD), dementia with Lewy bodies (DLB), or pure autonomic failure (PAF). The cases were classified, according to the guidelines for the pathological diagnosis of DLB, into three types: the neocortical type, the limbic type, and the brain stem-predominant type. All postmortem brains had coil-like glial inclusions as well as LBs, and the distribution pattern and density of glial inclusions and LBs varied. The distribution of glial inclusions was strikingly similar to that of LBs. In the cerebral cortex in particular, the number of glial inclusions was fairly well correlated with the number of LBs, irrespective of the three pathological types. In the brain stem, distribution was similar between glial inclusions and LBs, and there was no distinct pathological difference among the three types. Glial inclusions and LBs were immunohistopathologically similar with respect to ubiquitin, alpha-synuclein, and Gallyas-Braak staining. The clinical features of the three types of LB disease were also similar; i.e., parkinsonism, some dementia, and/or autonomic failure. The inclusions in neurons and glial cells occurred in parallel with respect to tissue distribution and immunohistochemical features, suggesting that accumulation of neuronal and glial inclusions in the LB diseases have a common pathological feature. Our findings suggest that DLB, PD with and without dementia, and PAF share one clinicopathological entity. PMID- 12624788 TI - Bismuth-induced neuronal cell death in rat dorsal root ganglion: a stereological study. AB - The goal of the present study has been to quantify the morphological changes in myelinated nerve roots and dorsal root ganglion (DRG) cells in rats exposed to bismuth subnitrate. Male Wistar rats (n =30) were divided into three groups of ten animals. The ten animals in each group were given intraperitoneal injections of one of the following: (1) 500 mg/kg bismuth subnitrate, (2) 1,000 mg/kg bismuth subnitrate, or (3) saline. The mean total cell number of B-cells in the DRG was significantly smaller in the two treated groups, 18% (2P <0.001) and 23% (2P <0.001), respectively, than it was in the control group. In addition, there was an 11% reduction in the number of A-cells (2P =0.039) in rats exposed to the highest concentration of bismuth. Bismuth did not affect the total number or mean cross sectional area of axons and myelin sheaths of the myelinated nerve fibers in the ventral or dorsal nerve root of the DRG. This is the first study to investigate pathological changes of the peripheral nervous system after bismuth intoxication. PMID- 12624789 TI - Expression of the CD34 antigen in pleomorphic xanthoastrocytomas. AB - Pleomorphic xanthoastrocytoma (PXA) is a rare, superficially located astrocytic glioma of children and young adults, which is associated with a relatively favorable prognosis. Here we report on the expression of the hematopoietic progenitor cell and vascular endothelial cell associated antigen CD34 in PXAs. We found CD34 immunoreactivity in various fractions of tumor cells in 44 of 60 PXAs investigated (73%). CD34 expression was more common in PXAs of WHO grade II (37 of 44 tumors, 84%) than in PXAs with anaplastic features (7 of 16 tumors, 44%). Immunoreactivity for CD34 was also commonly detected in single or clustered dysplastic neural cells within the cerebral cortex adjacent to the PXAs. Reverse transcription-PCR revealed that PXAs express the full-length CD34 transcript and a known splice variant encoding a truncated form of CD34. Both transcripts were detectable at higher levels in PXAs as compared to diffuse astrocytomas and non neoplastic brain tissue. Taken together, our findings demonstrate that PXAs frequently express CD34 not only in vascular endothelial cells but also in tumor cells and in dysplastic cells of the adjacent cortex. Therefore, immunostaining for CD34 may be a helpful tool for the histological differential diagnosis of PXAs. PMID- 12624790 TI - Peripheral neuropathy in systemic lupus erythematosus: pathomorphological features and distribution pattern of matrix metalloproteinases. AB - Matrix metalloproteinases (MMPs) are endoproteases that have been implicated in the pathogenesis of inflammatory and vasculitic neuropathies. In systemic lupus erythematosus (SLE), a peripheral neuropathy is frequently seen that is thought to be caused by ischemic nerve damage due to vasculopathy and/or vasculitis of the nutritional vessels. However, the exact pathomechanisms causing SLE neuropathy are largely unknown. Elevated MMP levels have been reported in the serum of SLE patients. Supposing that altered expression of MMPs may contribute to vessel wall damage in SLE neuropathy, we investigated the expression of MMP-1, -2, -3, -9, -10 and -13, and their tissue inhibitors (TIMP-1 and -2) in sural nerves from 12 SLE patients in comparison to normal controls. All MMPs could be detected within blood vessel walls from SLE nerves, whereas in controls MMP-3 and MMP-9 was not found. TIMP-1 and TIMP-2, on the other hand, were not informative. Generally, small and large nutritional vessels in the epineurium were immunoreactive for MMPs and TIMPs. Mononuclear cells, which expressed MMP-1, - 3, -10, -13, and TIMP-1 were also observed in most of the SLE nerves, mostly around epineurial blood vessels, but only occasionally in controls. This indicates that expression of MMPs in mononuclear cells may be related to leukocyte trafficking through the vessel walls. However, the density of TIMP-positive and MMP-positive inflammatory cells did not correlate with morphometric parameters regarding the severity of the neuropathy. Our findings suggest that especially the up regulation of MMP-3 and MMP-9 within the vessel walls may be responsible for the vascular damage seen in SLE and the resulting chronic combined axonal and demyelinating type of neuropathy frequently found in SLE. PMID- 12624791 TI - Abnormal trafficking of sarcolemmal proteins in alpha-glucosidase deficiency. AB - The dystrophin-associated protein complex (DAP) plays an important role in the integrity and stability of the muscle membrane. Whereas much is known about the interaction between DAP members at the sarcolemmal location, intracellular DAP assembly and trafficking is still largely unknown. In alpha-glucosidase (acid maltase) deficiency (alphaGDD), accumulation of glycogen is accompanied by cytoarchitectural abnormalities impairing normal protein metabolism. In the present study, we took advantage of this fact to examine the consequences of impaired protein handling on the formation of DAP, with the aim of gaining indirect knowledge about its sarcoplasmic trafficking and a better understanding of mechanisms leading to myopathic changes found in alphaGDD. Histological examination of alphaGDD muscle confirmed a vacuolar myopathy with glycogen accumulation both in vacuoles and within the sarcoplasm. Sarcoplasmic accumulation of sarcolemmal proteins, including dystrophin and sarcoglycans, occurred around some vacuoles and within non-vacuolated fibres. Utrophin was up regulated and found at extra-junctional sarcolemmal locations of many fibres. AlphaGDD muscle cells developed in a fashion similar to that of controls in culture. However, vacuoles were found in 2-week-old alphaGDD myotubes, and these subsequently increased in size and number. Substantial alterations in DAP handling were found, with accumulation close to the Golgi apparatus. Utrophin was not enriched in the sarcoplasm but was up-regulated along the whole sarcolemma. Our results demonstrate a close association of dystrophin and sarcoglycans during sarcoplasmic processing. Furthermore, they suggest that the myopathy found in alphaGDD is a secondary form of DAP deficiency. PMID- 12624792 TI - Role of protein kinase B in Alzheimer's neurofibrillary pathology. AB - Protein kinase B (PKB) is an important intermediate in the phosphatidylinositol-3 kinase signaling cascade that acts to phosphorylate glycogen synthase kinase-3 (GSK-3) at its serine 9 residue, thereby inactivating it. Activated GSK-3 has been previously shown to be preferentially associated with neurofibrillary tangles (NFTs) in Alzheimer's disease (AD) brain. In the present study, we performed immunohistochemistry with an antibody to the active form of PKB in brains with different stages of neurofibrillary degeneration. We found that the amount of activated PKB (p-Thr308) increased in correlation to the progressive sequence of AT8 immunoreactivity and neurofibrillary changes assessed according to Braak's criteria. By confocal microscopy, activated PKB (p-Thr308) was found to appear in particular in neurons that are known to later develop NFTs in AD. Western blotting showed that activated PKB was increased by more than 50% in the 16,000- g supernatants of AD brains as compared with normal aged and Huntington's disease controls. This increase in PKB levels corresponded with a several-fold increase in the levels of total tau and abnormally hyperphosphorylated tau at the Tau-1 site. These studies suggest the involvement of PKB/GSK-3 signaling in Alzheimer neurofibrillary degeneration. PMID- 12624793 TI - Origin and turnover of microglial cells in fibrillar plaques of APPsw transgenic mice. AB - Activated microglial cells are an integral component of fibrillar plaques in brains of subjects with Alzheimer's disease (AD) and in brains of transgenic mice overexpressing amyloidogenic fragments of human amyloid precursor protein (APP). The aim of this ultrastructural study of fibrillar plaques was to characterize the origin of microglial cells involved in cored plaque formation. Computer-aided three-dimensional reconstruction of plaques and microvessels in APPsw transgenic mice shows perivascular development of cored plaques. Perivascular location of almost all examined plaques and the infiltration at the interface between vessels and plaques with cells of monocyte/microglia lineage indicates that plaques are formed by inflammatory cells of blood origin. The increase in the number of microglial cells from 1 or 2 in an early plaque to more than 100 in a several month-old plaque does not result in plaque degradation, but is associated with amyloid core growth and progression of neuronal degeneration, and suggests that recruitment of inflammatory cells of blood origin sustains plaque growth. Infiltration of the plaque with cells of blood origin and degeneration of 10-46% of inflammatory cells in large plaques, which is especially frequent at the interface between capillary wall and plaque, suggest their accelerated turnover. PMID- 12624794 TI - Alpha-synuclein pathology affecting Bergmann glia of the cerebellum in patients with alpha-synucleinopathies. AB - We carried out immunohistochemical examinations of the brains (cerebella) of patients who had suffered from Parkinson's disease (PD), diffuse Lewy body disease (DLBD) or multiple system atrophy (MSA), using antibodies specific for alpha-synuclein. Alpha-synuclein-positive doughnut-shaped structures were found occasionally in the cerebellar molecular layer in some of these patients. Double labeling immunofluorescence and immunoelectron microscopy studies revealed that these alpha-synuclein-positive doughnut-shaped structures were located in the glial fibrillary acidic protein-positive radial processes of Bergmann glia, corresponding to the outer area of Lewy body-like inclusions, and consisted of granulo-filamentous structures. These findings indicate that, although not frequently, Bergmann glia of the cerebellum are also the targets of alpha synuclein pathology in alpha-synucleinopathies such as PD, DLBD and MSA. PMID- 12624795 TI - A systematic analysis of polyglucosan bodies in the human gastrointestinal tract in health and disease. AB - Polyglucosan inclusion bodies have been described in smooth muscle of the gastrointestinal tract of aged dogs, and rarely in association with enteric dysmotility in humans. We have systematically examined the human small and large bowel for the presence of such inclusions in health and motility disorders. Systematic, blinded, dual observer analysis of colonic and ileal tissue from patients (n=80, age 20-92 years) undergoing large bowel resections for non dysmotile conditions, principally neoplasia was performed, as well as retrospective review of all intestinal tissues referred for specialist histochemistry from patients undergoing surgery for motility disorders. All sections were stained with haematoxylin and eosin and periodic acid-Schiff stains. No polyglucosan bodies were identified in any specimen without dysmotility, regardless of age, but were a feature of 4/104 patients with diverse severe gastrointestinal motility disorders. In contrast to dogs, polyglucosan bodies are not a feature of normal ageing in human gastrointestinal smooth muscle but, in accord with previous suggestions, are seen in rare cases of human gut dysmotility. The significance of this difference is unclear. PMID- 12624796 TI - [Pulmonary hypertension in rheumatic diseases]. AB - Pulmonary hypertension (PH) is a progressive disease of the pulmonary vasculature characterized by increased vascular resistance and pressure overload of the right ventricle. Histologically, PH lungs demonstrate medial hypertrophy of small pulmonary arteries and proliferation of endothelial cells resulting in plexiform lesions. Recent studies have identified mutations of the bone morphogenetic protein receptor 2 (BMPR2) gene and the activin-receptor-like kinase 1 (ALK1) gene, that affect the transforming growth factor beta (TGF-beta) receptor superfamily, a group of transmembrane signaling molecules with serine-threonine kinase activity that are involved in the regulation of cell growth. Several lines of evidence indicate that the development of PH is a multi-hit process, where one of the events is having a gene mutation and another might be a circumstantial condition or other disease-modifying genes. It is unknown which mechanism that is critical in rheumatic diseases causes pulmonary vascular disease. PH is most frequently associated with systemic sclerosis (SS), systemic lupus erythematosus (SLE) and mixed connective tissue disease (MCTD), however, it is still a rare manifestation of these disorders. For example, approximately 10% of SS cases manifest pulmonary vascular disease. In recent years symptomatic vasodilator therapies have been employed and have been able to improve exercise capacity and survival in these patients. PMID- 12624797 TI - [Therapy of pulmonary involvement of rheumatic diseases]. AB - Pulmonary manifestations of rheumatologic diseases occur as an effect of immunological damage to the parenchyma and to the vasculature of the lung. Hence an intensive immunosuppressive management including the use of corticosteroids and cyclophosphamide as well as extracorporeal apheresis procedures has been suggested and found to be efficacious in small series of patients. Pulmonary hypertension secondary to inflammatory rheumatological disease is a well known complication of scleroderma and lupus. The new advent in the treatment of such conditions like bosentan or epoprostenol and their second generation followers opens new prospects towards the hitherto very dark prognosis for these patients. The most frequent pulmonary problem in rheumatological patients will be the increasingly frequent occurrence of microbial pneumonias due to aggressive therapeutic immunosuppression. PMID- 12624798 TI - [Histological classification of interstitial lung diseases]. AB - The 2002 ATS/ERS consensus classification of idiopathic interstitial pneumonias standardizes definitions and criteria for classification and diagnosis of idiopathic interstitial pneumonias and replaces previous classifications. Based on clinico-radiologic-pathologic criteria seven entities were defined: idiopathic pulmonary fibrosis, nonspecific interstitial pneumonia, cryptogenic organizing pneumonia, acute interstitial pneumonia, respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia and lymphoid interstitial pneumonia. The following paper includes a brief overview of the histopathological diagnosis of these entities as compared to other diffuse interstitial pulmonary diseases and pulmonary manifestations of collagenvascular diseases. PMID- 12624799 TI - [Imaging techniques in rheumatology: sonography in rheumatoid arthritis]. AB - Musculoskeletal ultrasonography has become an important diagnostic tool in rheumatoid arthritis. In Germany it is part of the rheumatology training, and many ultrasound courses provide further education. Only in the last five years the international importance of ultrasound in rheumatology has increased dramatically. Sonography can be performed as a bedside procedure and as an extension of the clinical investigation. It is easily tolerated by the patients, and it can be repeated any time. Sonography can have a great impact on therapeutic decisions. A > or = 5 MHz linear transducer is needed. Most transducers that are used for musculoskeletal ultrasound have about 7.5 MHz. Modern transducers with higher frequencies (>7.5 MHz) and high resolution improve the diagnostic value of the investigation. Sonography is superior to plain radiography to detect erosions as far as the region is accessible by ultrasound. It is more sensitive than the clinical investigation for the detection of synovitis, tenosynovitis, tendinitis, and bursitis as well as for the differentiation of these lesions. Color Doppler sonography aids in evaluating the activity of inflammation and in differentiating intraarticular structures. Carpal and ulnar neuropathy occur secondary to rheumatoid arthritis and may lead to characteristic nerve swelling. Ultrasound-guided injections into joints and tendon sheets can be performed. PMID- 12624801 TI - [The European perspective of rheumatology training]. PMID- 12624800 TI - [The rheumatologist's conflict between "off-label" prescription and failure to render assistance]. PMID- 12624802 TI - [5-HT3-receptor-antagonists in therapy of rheumatic diseases]. PMID- 12624803 TI - [The influence of depression on the effect of Tropisetron in the therapy of fibromyalgia]. AB - As described elsewhere the oral administration of 5 mg of the 5-HT3-receptor antagonist Tropisetron in fibromyalgia exhibited less amelioration of pain in patients with a depression in comparison to patients without depression. Since an intravenous treatment seems to increase the effect of Tropisetron, the question arises whether patients with depression profit from the intravenous therapy. Methods 68 out patients with fibromyalgia according to ACR-criteria were enrolled in the study. The patients filled in a VAS pain and the Beck Depression Inventory (BDI) before and after a bolus i.v. injection of 5 mg Tropisetron for 5 days [Beck AT, Steer Ra. Beck-Depression-Inventory (BDI) In: Hautzinger M (Hrsg der dt. Ausg.). Testhandbuch. 1. Auflage Bern: Verlag Hans Huber, 1994]. In the beginning the patients had to have > or = 40 mm in the VAS pain from 0-100 mm. The patients were divided into three groups: group 1 = patients with a BDI<19 without experience with antidepressive drugs (n=26); group 2=patients with a BDI > or = 19 (n=22) and negative experience with antidepressive substances, and group 3=patients with a BDI > or = 19 and an accompanying antidepressant drug therapy and some benefit under this therapy (n=20). Results Before the therapy there was no significant difference in VAS pain in the groups, but in BDI there was a significant difference between group 1 (BDI mean value 11.5) in comparison to group 2 (BDI mean value 26.1) and group 3 (BDI mean value 24.8). After therapy all three groups had a significant amelioration of pain: group 1: p=0.000023; group 2: p=0.00073; group 3: p=0.0145. There was a significant difference between the group with BDI<19 and the group with antidepressant drug in amelioration of pain (p=0.044). A significant correlation was found in group 2 with Beck > or = 19 between amelioration of pain and BDI after therapy (p=0.008, r=0.666). In this group a pain-reactive depression and in group 3 an endogenous depression must be discussed. PMID- 12624804 TI - [Sociodemographic characteristics, risk factors and reproductive history in subjects with fibromyalgia--results of a population-based case-control study]. AB - OBJECTIVES: According to recent studies, 1.3 to 4.8% of the population of Western industrialized nations suffer from fibromyalgia, with 80 to 90% of the affected persons being women. This preponderance of women, mostly in their reproductive years, suggests an association between fibromyalgia and sex hormones. METHODS: Within the framework of a population-based cross-sectional study of 3174 female residents of Bad Sackingen aged 35 to 74 years a stratified random sample of 653 women was clinically examined and surveyed. Thirty-six of these women fulfilled the ACR criteria of fibromyalgia. Forty-four women suffered from chronic widespread pain without having fibromyalgia (chronic pain group); 408 had no chronic pain (controls). RESULTS: Compared to the control group, subjects with fibromyalgia have a 3.6-fold risk, while compared to the chronic pain group, there is a 3.9-fold risk of having a lower social level. Subjects with fibromyalgia drink less alcohol than subjects from the chronic pain group and the control group. After adjusting for age, multivariate analyses demonstrated that, in comparison to controls, subjects with fibromyalgia have a significantly later menarche (OR=2.2 for >14 years) and had never been pregnant (OR=0.3). The comparison of the chronic pain group and controls did not demonstrate these associations. CONCLUSIONS: The associations with a low social level, low alcohol intake, late menarche and rare pregnancies are specific for subjects with fibromyalgia. These factors distinguish subjects with fibromyalgia from subjects with other chronic pain conditions as well as from subjects with no chronic pain. The same hormonal factors responsible for a delayed menarche and a reduced fertility may be relevant in the development of fibromyalgia. PMID- 12624805 TI - Measurement of disease activity in psoriatic arthritis. Extended report. AB - OBJECTIVE: To determine whether activity indices, generally accepted in rheumatoid arthritis (RA) are useful and valid to measure disease activity in psoriatic arthritis (PsA) patients with peripheral arthritis. METHODS: 38 PsA patients were studied before and after a one year DMARD treatment. Extended and reduced tender and swollen joint counts, Ritchie articular index, Health Assessment Questionnaire HAQ) score, erythrocyte sedimentation rate (ESR) morning stiffness, the patient's and the assessor's global assessment (PGA and AGA) were recorded. Disease activity scores, EULAR, ACR and Clegg improvement criteria were calculated. RESULTS: All indices correlated well before and after treatment with AGA (r > 0.337, p < 0.042), except morning stiffness and tender joint counts. After treatment, PGA correlated well only with the 68 and 28 tender joint counts, ESR and HAQ (r > 0.340, p < 0.05). The response to DMARD treatment was well characterized with the changes in the number of tender and swollen joint counts, and DAS4, DAS3, DAS28. The changes correlated with the PGA and AGA. The level of agreement between Clegg and the EULAR improvement criteria with both extended and reduced joint count was comparable (p < 0.01). CONCLUSION: The well-known activity indices generally accepted in RA, as tender and swollen joint count, DAS3, DAS4, DAS28, are useful and valid indices measuring arthritis activity in PsA with peripheral arthritis. The correlation between Clegg and EULAR improvement classification indices were similar. Both seemed to characterize changes authenticated during DMARD treatment. PMID- 12624806 TI - [Solitary tumorous muscular sarcoidosis (granulomatous myopathy) of the forearm extensors]. AB - INTRODUCTION: Besides the lung and hilar lymph nodes, sarcoidosis may affect almost every other organ. Under the aspect of histology, non-caseating granulomas are normally found in sarcoidosis but also appear in connection with other diseases. Therefore some authors prefer the definition "granulomatous myopathy" in the case of solitary muscular affection as long as sarcoidosis is not found in at least two other organs. Others, however, speak of "solitary muscular sarcoidosis", if any other origin can be excluded. CASE REPORT: A 20-year old man mentioned a painless swelling in his left forearm. During a 6-year follow-up, intermittent growth with long intervals of inactivity could be ascertained. The diagnosis was supported by two biopsies within three years. Under oral therapy with prednisone the tumor decreased evidently. REVIEW OF LITERATURE: Out of 61 cases of tumorous sarcoid myopathy described in literature, only 9 lesions appeared in just one location. Three of these 9 showed no evidence of disease in other organs and referred to lower limbs of Japanese. As far as we know a unilocular solitary tumorous sarcoid myopathy of the upper limb has not yet been described. CONCLUSION: The tumorous sarcoid myopathy is mainly located between the muscle fibers. Therefore paralysis, atrophy and contracture is rarely discovered in this kind of sarcoid myopathy. It shows a slow "stop-and-go" progress and may spontaneously go in remission. The expected success of therapy is therefore to be carefully considered in every incidence in view of side effects. Oral prednisone with an initial dose of 1mg per kg body weight per day seems to be the most effective therapy. A moderate dose reduction prevents recurrence. PMID- 12624807 TI - [Airway manifestations of relapsing polychondritis: treatment with cyclophosphamide and placement of bronchial stents]. AB - We report the case of a 41-year-old female patient with relapsing polychondritis and severe respiratory involvement. The patient presented with acute respiratory failure requiring endotracheal intubation. Bronchoscopy revealed tracheal collapse and inflammatory stenoses with dynamic collapse of the major airways. We describe a multidisciplinary therapeutic approach, consisting of immunosuppressive treatment, bronchoscopic placement of self-expandable stents into the collapsing bronchi, and tracheotomy. In addition, we report the effectiveness of oral cyclophosphamide for treatment of relapsing polychondritis with severe respiratory involvement after failure of other immunosuppressive agents. The problem of severe respiratory complications in patients with relapsing polychondritis and the need for a multidisciplinary approach is discussed. PMID- 12624808 TI - Spontaneous hemarthrosis of the knee associated with clopidogrel and aspirin treatment. AB - We report a case of a 76-year-old-man who developed spontaneous hemarthrosis of his right knee following clopidogrel-aspirin treatment. Clopidogrel is an ADP receptor antagonist and in combination with aspirin widely used in patients with atherosclerotic vascular disease to reduce the incidence of ischemic events. To date, no case of spontaneous hemarthrosis following clopidogrel-aspirin therapy has been reported. Prompt aspiration after discontinuing the ADP receptor antagonist-aspirin combination therapy can assist early diagnosis and may prevent further damage to the joint. In conclusion, spontaneous hemarthrosis is a possible complication following clopidogrel-aspirin therapy and is recommended to be evaluated when appropriate clinical symptoms (e.g., intraarticular effusion) present. PMID- 12624809 TI - [ANCA-associated systemic vasculitis (M. Wegener) with pulmonary manifestation]. PMID- 12624810 TI - [What's new in immunology?]. PMID- 12624811 TI - [The Greek and the Turk: Benediktos Adamantiades (1875-1965) and Hulusi Behcet (1889-1948)]. PMID- 12624812 TI - Recent development of antitumor agents from chinese herbal medicines; part I. Low molecular compounds. AB - A series of low molecular compounds from Chinese herbal medicines which have proved to be, in some cases, highly effective especially in tumor therapy, is listed here (part II will deal with high molecular compounds, to be published in the next issue). In contrast to synthetic agents used in cancer chemotherapy, these natural compounds have relatively low toxicities. Many of the clinical studies referred to in this paper have been carried out on Asians. Because genetic factors influence enzyme levels, sometimes leading to striking differences in metabolism and pharmacokinetics of drugs, results obtained in clinical studies carried out in China are not 100 % transferable to the European population. The mechanisms of action of these compounds are manifold, consisting of reactions with DNA bases, intercalation in DNA, inhibition of topoisomerases, inhibition of protein kinases, induction of apoptosis etc. Some of the compounds have interesting structural features, that may be used as lead structures for the development of further antitumor agents. PMID- 12624813 TI - Anti-human immunodeficiency virus type 1 (HIV-1) activity of lectins from Narcissus species. AB - Mannose-specific lectins (MSLs) were isolated from bulbs of fifteen wild Narcissus species growing in Spain and assayed for their HIV-1 infection inhibitory activity in MT-4 cells and compared to the Narcissus pseudonarcissus agglutinin (NPA), the commercially available MSL obtained from daffodils. Almost all the tested MSLs were found to be active, showing EC50 values (microg/mL) similar to that of NPA, with some being comparable to those obtained with dextran sulfate without significant cytotoxicity. However, on a molar basis almost all of the MSLs tested exhibited lower EC50 values than dextran sulfate whilst six MSLs had values lower than AZT. The most efficacious anti-HIV-1 activity was exhibited by the Narcissus tortifolious MSL, which was 10- (microg/mL) and 100- (molarity) fold more potent than dextran sulfate. Significantly, although this MSL was 15 fold less potent than AZT in terms of quantity (microg/mL), it was 68-fold more potent on a molar basis. The antiviral indices, a ratio of the concentrations that produce cytotoxicity and HIV-1 replication, were calculated and three of the MSLs, N. confusus, N. leonensis and N. tortifolius reported 1.5-, 2- and 8.5-fold greater AI values than dextran sulfate or AZT. Comparison of MSL haemagglutination activities (HAA) to their anti-HIV-1 activities showed that there was no significant correlation. It was suggested that this may be due to a dissociation between both activities as a consequence of multiple isolectin composition. PMID- 12624814 TI - Constituents of sage (Salvia officinalis) with in vitro affinity to human brain benzodiazepine receptor. AB - Benzodiazepine receptor binding assay-guided fractionation of the methanol extract from sage leaves ( Salvia officinalis L.) revealed three flavones and two abietane diterpenes functioning as benzodiazepine receptor-active components. Structural elucidation of the isolated pure compounds was performed by UV, EI-MS, ESI(pos)-MS/MS, as well as 1H- and 13C-NMR techniques. The flavones apigenin, hispidulin and cirsimaritin competitively inhibited 3H-flumazenil binding to the benzodiazepine receptor with IC50 values of 30, 1.3 and 350 microM, respectively. In addition, the affinities of the newly discovered diterpene receptor ligands, i. e., 7-methoxyrosmanol and galdosol, were characterized. 7-Methoxyrosmanol exhibited an IC50 value of 7.2 microM and galdosol showed the strongest binding activity to the benzodiazepine receptor with an IC50 value of 0.8 microM. PMID- 12624815 TI - The mechanism of inhibitory actions of S-petasin, a sesquiterpene of Petasites formosanus, on L-type calcium current in NG108-15 neuronal cells. AB - The effects of S-petasin, a sesquiterpene isolated from Petasites formosanus Kitamura, on ion currents in a mouse neuroblastoma and a rat glioma hybrid cell line, NG108-15, were examined with the aid of the whole-cell voltage-clamp technique. S-Petasin (1 - 300 microM) caused a decrease in the amplitude of L type Ca2+ current (I(Ca,L)) in a concentration-dependent manner, however, it did not change the overall shape of the current-voltage relationship of I(Ca,L). The IC50 value for S-petasin-induced inhibition of I(Ca,L) was 11 microM. S-Petasin (10 microM) shifted the steady-state inactivation of I(Ca,L) to a more negative membrane potential by approximately -10 mV. S-petasin could prolong the recovery of I(Ca,L) inactivation. The inhibitory effect of S-petasin on I(Ca,L) was found to exhibit tonic and use-dependent characteristics. S-Petasin could inhibit I(Ca,L) evoked by action potential waveforms effectively. S-Petasin also suppressed low voltage-activated I(Ca,L) in NG108-15 cells. S-Petasin at a concentration of 100 microM had little effect on voltage-dependent Na+ current; however, it did produce an inhibitory effect on delayed rectifier K+ current in a time-dependent manner. These results demonstrate that S-petasin can interact directly with L-type Ca2+ channels in NG108-15 cells. These effects could contribute to the regulation of neuronal activity if similar results were found in neurons in vivo. PMID- 12624816 TI - The depressor and vasodilator effects of rutaecarpine are mediated by calcitonin gene-related peptide. AB - Previous studies have shown that rutaecarpine has depressor and vasodilator effects, and activates vanilloid receptors to evoke calcitonin gene-related peptide (CGRP) release. In the present study, we examined whether the depressor and vasodilator effects of rutaecarpine are related to the stimulation of endogenous CGRP release via activation of vanilloid receptors in rats. Rutaecarpine (30, 100, or 300 microg/kg, i. v.) caused a depressor effect concomitantly with an increase in the plasma concentrations of CGRP in a dose dependent manner, and the effects of rutaecarpine were abolished by pretreatment with capsaicin (50 mg/kg, s. c.) which depletes neurotransmitters in sensory nerves. In aortic and superior mesenteric arterial rings, rutaecarpine (10 (-7) 10(-5) M) or capsaicin (3 x 10(-9)-3 x 10(-6) M) caused a concentration-dependent vasodilator response, which was significantly attenuated by capsazepine (10(-5) M), a competitive vanilloid receptor antagonist, or by CGRP-(8-37) (10(-6) M), a selective CGRP receptor antagonist. After pretreatment with capsaicin (10(-5) M) for 20 min, vasodilator responses to rutaecarpine were also markedly attenuated. Similarly, pretreatment with rutaecarpine (10(-5) M) for 20 min also attenuated vasodilator responses to capsaicin. These results suggest that the depressor and vasodilator effects of rutaecarpine are related to stimulation of endogenous CGRP release via activation of vanilloid receptors in rats. PMID- 12624817 TI - Honokiol ameliorates cerebral infarction from ischemia-reperfusion injury in rats. AB - Honokiol, a constituent extracted from Magnolia officinalis, had been shown be an antioxidant and an anti-platelet agent in biological systems with an anti arrhythmic effect and a myocardial protective effect on ischemia-reperfusion injury. We examined the neuroprotective effect of honokiol in rats subjected to focal cerebral ischemia. Honokiol was administered intravenously either at fifteen minutes before right middle cerebral artery occlusion (pretreatment groups) or when both common carotid arteries clips were removed (treatment groups). The results showed that there was no significant hemodynamic change after intravenous infusion of honokiol at the dosages of 10(-8), 10(-7) and 10( 6) g/kg in both groups. However, honokiol significantly reduced the total volume of infarction at the doses of 10(-7) or 10(-6) g/kg in both pretreatment and treatment groups. This study suggests that honokiol is a potent neuroprotective agent in focal cerebral ischemia. This beneficial effect may be related to its antioxidant effect and anti-platelet aggregation activity. PMID- 12624818 TI - Pharmacological effects of (+)-nantenine, an alkaloid isolated from Platycapnos spicata, in several rat isolated tissues. AB - In this work, the potential activity of (+)-nantenine (a natural aporphin alkaloid) in several rat isolated tissues was studied. In rat isolated intact aorta, (+)-nantenine (0.05 - 0.5 microM) competitively antagonized with almost equal effectiveness the contractions produced by phenylephrine (PE) and 5 hydroxytryptamine (5-HT) in normal Krebs solution. However, at higher concentrations (2 microM), the alkaloid also reduced the maximal effect induced by these two agonists. In depolarizing Ca2+-free high KCl 50 mM solution, (+) nantenine (1.5 - 6 microM) inhibited, in a non-competitive way, the increase in tension evoked by Ca2+ with depression of the maximum response. On the other hand, (+)-nantenine (3 - 30 microM) did not affect the contractile effect caused by okadaic acid (OA, 1 microM) while, however, this alkaloid totally relaxed, in a concentration-dependent fashion, the contractions produced by phorbol 12 myristate 13-acetate (PMA, 1 microM) in endothelium-containing rat aortic rings. (+)-Nantenine (1 - 30 microM) reversed and competitively antagonized the inhibitory action induced by B-HT 920 in electrically-stimulated rat vas deferens. In isolated rat atria, (+)-nantenine (3 - 10 microM) diminished the contraction frequency. (+)-Nantenine (3 microM) significantly reduced the depolarization (voltage)-activated transient (T-type) and sustained (long lasting, L-type) barium inward currents [ IBa(T) and IBa(L) ] recorded in whole cell-clamped rat aortic myocytes. These results indicate that the pharmacological effects of (+)-nantenine observed at concentrations lower than 1 microM can be attributed to alpha 1 -adrenergic and 5-HT 2A receptor blocking properties whereas at higher concentrations (> 1 microM) the pharmacological activity of this natural compound may be also due to a decrease of Ca2+ influx through transmembrane calcium channels (calcium antagonist activity), to an inhibition of PKC actions and/or to an alpha 2 -adrenoceptor blockade. PMID- 12624819 TI - Agrobacterium tumefaciens-mediated genetic transformation of the cardenolide producing plant Digitalis minor L. AB - A repeatable transformation system has been established for Digitalis minor using Agrobacterium tumefaciens. Leaf explants from 30-day-old seedlings were inoculated with either EHA105 (carrying the nptII and gusA genes) or AGL1 (with the bar and gusA genes) strains. Among the tested factors influencing T-DNA transfer to plants, the EHA105 strain and the addition of acetosyringone to the co-culture medium increased transformation. The highest transformation efficiency (8.4 %) was obtained when freshly isolated explants, soaked in a bacterial suspension with an OD550 of 0.9, were subcultured on selection medium after a 4 day co-culture with the bacteria. Evidence of stable transgene integration was obtained by PCR, growth on media selective for nptII or bar genes, and expression of the gusA gene. Southern hybridisation, performed in six plants, provided information about the number of inserts. More than 200 transgenic plants were recovered from 65 independent explants. Thirty of these plants were successfully established in soil. This is the first report on transgenic Digitalis spp plants using an A. tumefaciens-mediated leaf disc transformation procedure. PMID- 12624820 TI - Two new protopines argemexicaines A and B and the anti-HIV alkaloid 6 acetonyldihydrochelerythrine from formosan Argemone mexicana. AB - Two new protopine-type alkaloids, argemexicaine A (1) and argemexicaine B (2), along with thirteen known alkaloids, were isolated from MeOH extracts of Formosan Argemone mexicana L. (Papaveraceae). Physical and spectral analyses, particularly IR and thermo-modulated 1D and 2D NMR, were used to determine the transannular conformations of the isolated protopine-type alkaloids. The known benzo[ c]phenanthridine (+/-)-6-acetonyldihydrochelerythrine (5) exhibited significant anti-HIV activity in H9 lymphocytes with EC50 and TI (Therapeutic Index) values of 1.77 microg/mL and 14.6, respectively. PMID- 12624821 TI - In vitro anti-influenza virus activity of isoquinoline alkaloids from thalictrum species. AB - The in vitro anti-influenza virus effects of some isoquinoline alkaloids, isolated from Thalictrum species (Ranunculaceae), growing in Mongolia and Sweden have been studied. (-)-thalimonine (Thl) and (-)-thalimonine N-oxide, isolated from the Mongolian plant T. simplex, inhibited markedly the influenza virus reproduction in vitro; thalictuberine N-oxide was less effective. The synthetic analogues of Thl as well as the rest of the tested alkaloids, isolated from T. foetidum and T. flavum, were not active. At a concentration range between 0.1 - 6.4 microM Thl inhibited viral reproduction in a selective and specific way. PMID- 12624822 TI - Coumarins and carbazoles from Clausena excavata exhibited antimycobacterial and antifungal activities. AB - Four known coumarins, dentatin (1), nor-dentatin (2), clausenidin (3) and xanthoxyletin (5), and six known carbazole derivatives, 3-formylcarbazole (6), mukonal (7), 3-methoxycarbonylcarbazole (8), murrayanine (9), 2-hydroxy-3-formyl 7-methoxycarbazole (10) and clauszoline J (11) were isolated from Clausena excavata. Compounds 1 and 6 were first isolated from the crude chloroform extract of the rhizomes. Compounds 1, 2, 3, 6, 7, 8, 10 and 11 showed antimycobacterial activity at a minimum inhibitory concentration (MIC) of 50, 100, 200, 100, 200, 50, 100 and 100 microg/mL, respectively. O-Methylated clausenidin ( 4), prepared from 3, exhibited antimycobacterial activity at MIC 50 microg/mL. Compounds 6, 7, 8 and 10 showed antifungal activity with IC 50 values of 13.6, 29.3, 9.5 and 2.8 microg/mL, respectively. All compounds demonstrated no cytotoxicity against KB and BC-1 cell lines. PMID- 12624823 TI - Composition and antimicrobial activity of the essential oil of Artemisia absinthium from Croatia and France. AB - The essential oils obtained by steam distillation from the aerial parts of two populations of Artemisia absinthium, from France and from Croatia, were analyzed by GC and GC-MS. The oils of A. absinthium of French origin contain (Z) epoxyocimene and chrysanthenyl acetate as major components while the oils of Croatian A. absinthium contain mainly (Z)-epoxyocimene and beta-thujone. Analysis of oils before and after anthesis showed some quantitative differences. Analysis of separated leaves and flowering heads showed only few differences among these organs. As they contain no thujone, antimicrobial screening was performed on samples of French origin and showed that A. absinthium oil inhibited the growth of both tested yeasts (Candida albicans and Saccharomyces cerevisiae var. chevalieri). PMID- 12624824 TI - In vitro studies on the mechanism of action of two compounds with antiplasmodial activity: ellagic acid and 3,4,5-trimethoxyphenyl(6'-O-aalloyl)-beta-D glucopyranoside. AB - To investigate the mechanism of action of two antiplasmodial compounds, ellagic acid and 3,4,5-trimethoxyphenyl (6'-O-galloyl)-beta-D-glucopyranoside (TMPGG), we studied in vitro two metabolic reactions of intraerythrocytic parasites: the activity of recombinant plasmepsin II, one of the haemoglobin proteases, and the detoxification of haematin into beta-haematin. Both compounds inhibited plasmepsin II activity, but at concentrations ten-fold higher than those needed for inhibiting parasite growth. Moreover, ellagic acid inhibited the formation of beta-haematin, with an IC50 only 3-fold higher than that of chloroquine. These data suggest that the antiplasmodial activity of ellagic acid could be related to the inhibition of beta-haematin formation, whereas plasmepsin II does not represent the main target of the two compounds. PMID- 12624825 TI - In vitro cytotoxic elemanolides from Vernonia lasiopus. AB - Two new elemanolides, epivernodalol and lasiopulide, were isolated after chromatographic separation of the alcoholic extract of the dried aerial parts of the Vernonia lasiopus. These elemanolides are new C-10 epimers of the sesquiterpene lactones vernodalol and demethylacroylated vernodalol isolated from other species of Vernonia. Both elemanolides showed in vitro cytotoxicity against human cancer cell lines in culture. This is the first report of isolation and cytotoxic activity of the two elemanolides from V. lasiopus. PMID- 12624826 TI - Cytotoxic activity of natural labdanes and their semi-synthetic modified derivatives from Croton oblongifolius. AB - Labda-7,12( E),14-triene-17-oic acid, previously isolated from Croton oblongifolius, and its derivatives were investigated for cytotoxicity against five human tumor cell lines. Six of these compounds, labda-7,12( E),14-triene-17 al, 17-hydroxylabda-7,12( E),14-triene, 17-acetoxylabda-7,12( E),14-triene, 15 hydroxylabda-7,13( E)-diene-17,12-olide, labda-7,13( E)-diene-17,12-olide, and 12,17-dihydroxylabda-7,13( E)-diene showed non-specific, moderate, cytotoxicity against all cell lines, whereas the other compounds were inactive. PMID- 12624827 TI - Pulsatilloside A and anemoside A3 protect PC12 cells from apoptosis induced by sodium cyanide and glucose deprivation. AB - Using sodium cyanide (NaCN) and glucose deprivation induced cell injury in PC12 as an injury model, we investigated the protective effects of pulsatilloside A and anemoside A 3 on neurons. The results showed that PC12 cells under the NaCN injury and glucose deprivation would undergo apoptosis. Additions of pulsatilloside A and anemoside A 3, at dosages ranging from 0.1, 1 and 10 microg/ml, protected PC12 cells from apoptosis determined by MTT, LDH release analysis, and flow cytometry measurement. PMID- 12624829 TI - Jatrophane diterpenes from the latex of Euphorbia obtusifolia with inhibitory activity on the mammalian mitochondrial respiratory chain. AB - Seven diterpenes isolated of the latex of the Euphorbia obtusifolia were evaluated as inhibitors of the NADH oxidase activity in submitochondrial particles from bovine heart. Compound 2, 2,3,5,7,8,9,15-heptahydroxyjatropha 6(17),11-diene-14-one 8,9-diacetate 7-isobutyrate 2,3-bis(2-methylbutyrate), was the most potent inhibitor with an inhibitory concentration (IC 50 ) value of 5.1 +/- 0.2 microM. In the present study, some structure-activity trends are suggested for the inhibitory activity of these natural products on the mammalian mitochondrial respiratory chain. PMID- 12624828 TI - Clionasterol: a potent inhibitor of complement component C1. AB - Clionasterol (1a), clionasterol monoacetate (1b) and 5alpha,8alpha-epidioxy 24alpha-ethylcholest-6-en-3-ol (2), isolated from the marine sponge Xestospongia exigua, and beta-sitosterol (3) were tested for their influence on the classical (CP) and alternative (AP) pathways of activation of the human complement system in vitro. All the sterols inhibited the CP in a dose-dependent manner but no detectable effect was observed in the AP even at concentrations of 400 microM. Clionasterol was found to be a potent inhibitor of CP (IC50 = 4.1 microM) being ten-fold more active than beta-sitosterol. The presence of the epidioxy group on C-5 and C-8 of compound 2 caused a pronounced decrease of the inhibitory effect. Mechanistic studies on the anticomplementary effect of clionasterol revealed that it interferes with the complement component C1. PMID- 12624830 TI - Vasorelaxing properties and bio-guided fractionation of Cedrelopsis grevei. AB - The activity-guided fractionation of the hydroalcoholic extract of trunk bark of Cedrelopsis grevei led to the isolation of five coumarins, norbraylin (1), methyl O-cedrelopsin (2), cedrecoumarin A (3), scoparone (4) and braylin (5) as a part of the constituents responsible for the vasorelaxing activity observed for the crude extract. PMID- 12624831 TI - Sesquiterpene lactones from Taraxacum obovatum. AB - Two new guaianolide glucosides, deacetylmatricarin 8-O-beta-glucopyranoside and 11beta-hydroxyleukodin 11-O-beta-glucopyranoside, were isolated from roots of Taraxacum obovatum, along with four known sesquiterpene lactones, deacetylmatricarin, sonchuside A, taraxinic acid beta-glucopyranosyl ester and its 11beta,13-dihydro derivative. Their structures were established by spectral methods. PMID- 12624832 TI - Inhibition of mutagenicity of food-derived heterocyclic amines by sulphoraphene- an isothiocyanate isolated from radish. AB - The naturally derived isothiocyanate, sulphoraphene [4-isothiocyanato-(1R) (methylsulphinyl)-1-(E)-butene], isolated from seeds of radish ( Raphanus sativus L., Cruciferae) was investigated for its antigenotoxic effects against a battery of cooked food mutagens (heterocyclic amines) in the Ames Salmonella/reversion assay using Salmonella typhimurium TA98 (frame-shift mutation sensitive) and TA100 (base -pair mutation sensitive) bacterial strains in the presence of Aroclor 1254 induced rat liver S9. Results of the present in vitro anti mutagenicity studies using the base-pair mutation sensitive strain TA100, strongly suggest that sulphoraphene is a potent inhibitor of the S9-mediated mutagenicity of all the tested heterocyclic amines (60 - 75 % inhibition at a dose of 500 nmol/plate). PMID- 12624833 TI - Identification of bulb from Fritillaria cirrhosa by PCR with specific primers. AB - Bulb of Fritillaria cirrhosa is an important traditional Chinese herbal medicine. According to the Chinese Pharmacopoeia (1995), it is commonly used as an antitussive and expectorant. Many young bulbs from species of Fritillaria are similar to those of F. cirrhosa, but they are different in price and quality. Therefore, there are many young bulbs from species of Fritillaria that could fake those of F. cirrhosa on the commercial market. The coding region of 5S-rRNA is highly conserved in higher eukaryotes. The 5S-rRNA spacer region sequences of F. thunbergii, F. pallidiflora, F. ussuriensi, F. delavayi, F.cirrhosa, F. anhuiensis, F. puqiensis were cloned by PCR with a pair of primers located within the conserved coding region. Based on sequences analyses of the 5S-rRNA spacer region from the 7 species, a specific sequence was found in F. cirrhosa. A pair of specific primers was designed for differentiating the bulbs of F. cirrhosa from each other by PCR. This result indicated that the method is rapid, more accurate and applicable in identification of the bulbs of F. cirrhosa at the DNA level. PMID- 12624834 TI - [On the diagnostics of structural deficits in dyslexia in clinical work: relationship between different achievement indicators]. AB - BACKGROUND: The aim of the study is to research the relationship between different indicators used in the diagnosis of reading and writing disabilities in clinical work. METHODS: In addition to being tested for intelligence and writing ability, a total of 141 children in second and fourth grade were submitted to tests assessing their ability for speech intelligibility in noise, dichotic listening, phoneme discrimination, as well as auditory short-term memory for digits and non-words. In a partial sample of 40 children with average intelligence manifesting at least a standard deviation between their ability to write and their IQ a regression analysis was introduced to assess the predictive power of applied indicators for writing ability. RESULTS: Next to IQ testing, phoneme analysis as used in the Heidelberg Test of Phoneme Discrimination accorded for the variance in writing ability. However, the relationship between these indicators proved to be most important for the beginning of learning the written language. CONCLUSIONS: The diagnostic procedures used in clinical work are critically evaluated whereby the tests to assess speech intelligibility in noise and dichotic listening, as well as short-term memory appear to be clinically relevant too, however a standardized procedure and age-relevant norms are still needed. PMID- 12624835 TI - [Acoustic schwannoma with progressive hearing loss in children. A case report]. AB - BACKGROUND: Neurinomas of the vestibulocochlear nerve unrelated to neurofibromatosis in children are extremely rare. Only 20 cases in children under the age of 16 are reported in the literature. Progressive unilateral hearing loss, tinnitus, vertigo and neurological deficits due to cranial nerve or brainstem compression are clinical signs. PATIENT AND RESULTS: We report on the case of a 12-years-old girl with an unilateral hearing loss, progressing to total deafness. Otoacoustic emissions were normal. In the MRI a large cerebellopontine angle tumor was found, identified as schwannoma of the vestibulocochlear nerve. CONCLUSIONS: The importance of MR Imaging in children with progressive unilateral hearing loss is demonstrated in this case. PMID- 12624836 TI - [Aural fullness and endolymphatic hydrops]. AB - BACKGROUND: Low-frequency modulated DPOAEs were registrated to investigate whether the subjective symptom of aural fullness can indicate an endolymphatic hydrops. METHOD: The cochlear partition is periodically moved towards scala vestibuli and scala tympani by a low-frequency suppressor tone. The level of simultaneously registrated DPOAEs is modulated depending on the phase of the suppressor. This modulation may be reduced when the displacement of the organ of Corti is inhibited by mechanical stiffening of the basilar membrane (e. g. in endolymphatic hydrops). SUBJECTS: Low-frequency modulated DPOAEs were registrated in 15 patients with Meniere's disease, in 8 patients with aural fullness and tinnitus, but without vertigo, and in 21 normal hearing adults. RESULTS: In both patient groups the modulation depth is high significantly lower than in the control group. The results do not differ significantly in the patients with and without vertigo. CONCLUSION: The reduction of the modulation depth can be interpreted as a sign of an endolymphatic hydrops. In the group of patients with aural fullness a cochlear hydrops can be assumed. The measurement of low frequency modulated DPOAEs is a new tool to diagnose and monitor the course of endolymphatic hydrops and to evaluate the effectiveness of therapeutic methods. PMID- 12624837 TI - [Early postoperative middle-ear ventilation -- risk for the transplant or guarantee for aeration of the tympanic cavity?]. AB - BACKGROUND: Sufficient ventilation of the tympanic cavity is a decisive factor for the success of the therapy and the prognosis after tympanoplastic surgery. With the postoperative use of the Valsalva- and Politzer-maneuver the patient is able to influence the aeration of the tympanic cavity in a positive way by autoinsufflation. However this implicates the risk of a dislocation of the transplant, which is possibly caused by high middle-ear pressure-peaks during the maneuvers mentioned. METHODS: The actual increase of pressure in the tympanic cavity respectively auditory canal during the Valsalva- and Politzer-maneuver is examined in 22 patients with chronic-mesotympanic Otitis media and additionally by using an experiment with a model of a reconstructed middle-ear. The risk of a transplant-dislocation after tympanoplasty type I is estimated. RESULTS: The increase of pressure in the middle ear respectively auditory canal -- and with that the possibility of a dislocation of the transplant -- is lower after Politzer- compared with Valsalva-maneuver. If the transplant is secured through silicon sheets and packings in the auditory canal, the probability of "blowing away" the newly created cover of the tympanon will be very small. CONCLUSIONS: Because of the benefits for the aeration of the tympanic cavity, an early post operative insufflation of the middle-ear -- using the Politzer- and/or "cautious" Valsalva-maneuver -- is recommended. PMID- 12624838 TI - [Naphazoline intoxication: course and therapy]. AB - BACKGROUND: As potent alpha-adrenergic agonists imidazoline derivatives like naphazoline are frequently used drugs because of their vasoconstrictive and decongestant properties. Beside their well-known local side effects also rare systemic ones like hypertension, excitability and nausea are possible. In case of intoxication this can enhance to ischemia of vital organs, bradycardia with arrhythmia, respiratory depression, hypothermia, excitation or marked sedation. PATIENTS: Two cases of intoxication after application of naphazoline for haemostasis under the operation, the postoperative course and therapy are reported. RESULTS: No specific antidote is available but a symptomatic drug therapy with intravenous administration of 5 mg phentolamine, an alpha adrenoceptor antagonist acts against the vasoconstrictive side effects. CONCLUSIONS: Because of the serious side effects and the narrow therapeutic to toxic window of naphazoline a strict observance of guidelines and contradictions, such as maximum dose, is necessary. PMID- 12624839 TI - [NPY in human nasal mucosa -- an immunocytochemical and immunoelectron microscopical study]. AB - BACKGROUND: The functions of the nasal mucosa are regulated by numerous endogenous and exogenous influences. The innervation patterns are important for the control of the physiological nasal functions. In addition to the classic neurotransmitters different neuropeptides might play a regulating and modulating role in the nasal mucosa. Both the significance and the localization of neuropeptide Y (NPY) have not been fully elucidated. PATIENTS AND METHODS: Tissue samples of human inferior turbinates from 42 patients were taken during nasal surgery and preserved in phosphate-buffered paraformaldehyde or glutaraldehyde. Serial sections were incubated with antibodies against NPY and the ABC method was applied. In order to identify immunoelectron microscopic reactions a streptavidin gold-marker was used. The findings were photodocumented by using a light- and transmission-electron microscope. RESULTS: NPY-positive terminals were mainly located in the adventitia of arterial vessels. There were also NPY-immunoreactive arterioles near to the glands. Periglandular a lower density of immunoreactions could be observed. NPY-positive fibers could be detected in the subepithelial connective tissue and at the glandular ducts. Immunoelectron microscopy revealed NPY within periglandular axons. CONCLUSIONS: Immunohistochemical and immunoelectron microscopical methods allow a detailed identification of the sympathetic cotransmitter NPY in arterial vessels of nasal mucosa in man. These results indicate that NPY-containing nerve fibers innervate arteries as well as nasal glands. These findings suggest that NPY play a significant role as a neuromodulator in the control of both vasculature and glandular secretion. The localisation of NPY in periglandular and periductal nerves confirms the direct influence of glandular functions. NPY-agonists may be a beneficial additional treatment of rhinopathies to reduce nasal obstruction and mucus secretion. PMID- 12624840 TI - [First experiences in endonasal duraplasty without antibiotics]. AB - BACKGROUND: Available evidence does not support the use of prophylactic antibiotics in patients with CSF fistulae. The question arises whether an antibiotic prophylaxis or therapy is mandatory planning an operative closure of frontobasal dural lesions. PATIENTS: In a retrospective survey a consecutive sample of 7 patients who had undergone endonasal ENT-department of two universities and an university teaching hospital was taken. The hospital course was evaluated and all patients were interviewed after a postoperative period between 6 and 36 months. Endonasal duraplasty by underlay or onlay technique without pre- or intraoperative application of antibiotics was performed. Hospital course, development of meningitis, need for antibiotic therapy later on, subjective complaints, history of meningitis, cerebrospinal fluid rhinorrhea, fluoresceine test (intrathecal administration of fluoresceine sodium and subsequent nasal endoscopy) were evaluated. RESULTS: 6 patients had an uneventful postoperative hospital course without any sign of inflammatory complications. In one patient with a larger resection of dura antibiotics were administered at the second postoperative day because of recurrent fever. There occurred no meningitis in any patient. All duraplasties were successfully closed. 5 patients showed a negative fluoresceine test, 2 patients denied it having no special problems. CONCLUSIONS: Duraplasty can be performed satisfactorily by endonasal approach. In small lesions and in the absence of special risk factors (diabetes, acute sinusitis, immunodeficient patients) endonasal duraplasty don't seem to inevitably need an antibiotic cover. A larger patient population is necessary to confirm these results. PMID- 12624841 TI - [Gastroesophageal reflux -- a common illness?]. AB - For a long time heartburn was not considered a symptom for serious illness. By now, however, it is accepted that the incidence of secondary carcinoma of the esophagus caused by chronic GERD has increased dramatically since the nineteen seventies. Mechanisms leading to GERD are complex and its incidence is not necessarily pathological. However pathological reflux in the lower esophagus (pH lower than 4 in 6 % of 24 hours), caused by decreased sphinctertonus, impaired peristalsis and clearance of the esophagus, may lead to complications. Helicobacter pylori may play a key role in GERD. There is strong evidence for a protective effect of Hp-infection in the development of GERD. In pangastritis, caused by Hp-infection, gastric acid production is inhibited resulting in a reduction of stomach-acid-concentration. This may be caused by either the chronic infection itself and the resulting atrophy of the stomach-mucosa, by the ammonia producing HP-bacteria, or an increase in acid re-absorbtion of gastric epithelium. Laryngopharyngeal reflux (LPR) often results in atypical manifestations with oral, pharyngeal, laryngeal, and pulmonary disorders. Laryngopharyngeal reflux is known to contribute to posterior acid laryngitis and laryngeal contact ulceration or granuloma formation, laryngeal cancer, chronic hoarseness, pharyngitis, asthma, pneumonia, nocturnal choking, and dental diseases. Today, PPI are the medication of choice in both acute and long-term (prophylactic) therapy of GERD. The so called "step-up-strategy" of medication is no longer recommended. Here, patients were first treated with antacids, then prokinetics followed by H2-blockers and finally low-dose PPI. Only in the case of persisting symptoms medication was further increased to high-dose PPI therapy. In the past this increase in medication lead to a prolonged healing process and consequently to higher medication costs. Studies have shown that a "step-down" therapy, beginning with high dose PPI, is highly preferable, since it is much more effective. Depending on the degree of the symptoms, however, medication may also be applied "on-demand". The BfArM has approved this kind of medication application only for Esomeprazol (Nexium mups 20 mg). PMID- 12624842 TI - [Surgical treatment of severe velopharyngeal insufficiency following uvulopalatopharyngoplasty (UPPP)]. AB - BACKGROUND: For the implementation of an uvulopalatopharyngoplasty (UPPP) it is nowadays common practice to operate in a manner which is gentle on the tissue and which retains as much muscle as possible. However, even when the greatest possible care is taken during the operation, serious complications such as persistent velopharyngeal insufficiency can arise. PATIENT AND METHOD: Based on a case report of a 55 year old man with a persistent severe velopharyngeal insufficiency after UPPP performed at another institution and serious OSA, we suggest a so-called push back technique, adapted from the cleft-palate surgery, for the extension of the soft-palate. With regard to the operative technique, it is thus possible to produce an effective extension of the soft-palate by means of a dorsocaudal displacement of a wide and on the palatal vascular fascicle pedicled mucoperiosteum flap of the mucous membrane of the hard palate, as well as similar mobilisation of the soft-palate from the os-palatinum. RESULT: Only 3 weeks after the operation, our patient showed a clinically sufficient velopharyngeal functioning. After 3 months, a complete restoration of the nasal CPAP-acceptance was evident. As a result, a complete social rehabilitation was possible for our patient. CONCLUSION: The modified push back technique of the palate retrodisplacement which is described above offers an efficient possibility of anatomically and functionally correcting the persistent and serious velopharyngeal insufficiency after UPPP. PMID- 12624843 TI - [Preliminary experiences with Photopic ultrasound imaging in the head and neck region]. AB - BACKGROUND: The application of high end ultrasound platforms increases the quality of sonographic images. New software developments like Tissue Harmonic Imaging (THI) have improved the resolution of images and help to better visualize anatomic findings. Photopic-mode uses color-coding to gain contrast. METHODS: This investigation compares the use of the Photopic-software in comparison to conventional B-mode. The ultrasound platform Sonoline Elegra was used. RESULTS: The Photopic-mode revealed in 8 of 40 patients a slight improvement within image quality. However, the use of Photopic-mode did not lead to other diagnostic results. CONCLUSION: Ultrasound investigation with Photopic-mode can be a useful supplement to the conventional B-mode-sonographic examination in selected cases by gain in contrasting. A disadvantage of the Photopic-mode is that it cannot be used simultaneously with color Doppler-mode examination. PMID- 12624844 TI - [Surgical treatment of rhinophyma with the ultrasonic scalpel (Ultracision Harmonic Scalpel)]. AB - BACKGROUND: There are multiple surgical approaches to the treatment of rhinophyma. Because of the exceptional vascularity of the nose, traditional surgical approaches to the management of rhinophyma are fraught with persistent intraoperative hemorrhage. Hemorrhage that is controlled with electrocautery or laser by a combination of excision of rhinophymatous tissue and simultaneous hemostasis invariably damages cartilage by thermal injury. The UltraCision Harmonic Scalpel achieved a tissue dissection with a simultaneous hemostasis and only minimal thermal tissue injury. To this date, in Europe no studies have been published on the use of the Ultracision Harmonic Scalpel in the surgical treatment of rhinophyma. PATIENTS AND TREATMENT: 6 patients had surgical treatment of rhinophyma with the UltraCision Harmonic Scalpel. The objective of the study was therefore to analyse the handling indication, the efficiency of intra- and postoperative hemostasis, the wound healing, the postoperative pain and the histologic results of tissue resections. RESULTS: Because of the effective intraoperative hemostasis supplementary hemostasis was not required in any examined patient. The efficient hemostasis and the tactile experience of the UltraCision Harmonic Scalpel enabled an accurate sculpting of the nose. Postoperative pain sensation was minimal. Wound healing was uneventful in all cases. Postoperative reepithelialization occurred within 28 days. Delayed complications like scarring did not occur within a median follow - up of 13 month. Histopathological evaluation was possible in all cases. CONCLUSION: The UltraCision Harmonic Scalpel offers a surgical treatment of rhinophyma with efficacious intra- and postoperative hemostasis. The combination of simultaneous tissue dissection and hemostasis enables a good overall view and control of the surgical site. Based on its mechanical function, the UltraCision Harmonic Scalpel has only a little thermal effect on neighbouring tissues and a good tactile experience that enables a controlled tissue resection. The UltraCision Harmonic Scalpel combines the advantages of tissue dissection and simultaneous hemostasis with the controlled handling of the traditional scalpel in rhinophyma surgery. PMID- 12624845 TI - [The interesting case -- case no. 55. Median dermoid cyst of the tongue]. PMID- 12624846 TI - [Construction and reaction of the mucosa-associated-lymphatic tissue (MALT) in the middle ear, the Tuba Eustachii and the larynx of the rat]. PMID- 12624847 TI - [Pros and cons as to ICD-10 chapter V]. PMID- 12624848 TI - [Burden of relatives and their expectations towards psychiatric institutions. A review of the literature and own results]. AB - OBJECTIVE: This article aimed to provide an overview regarding the burden of relatives of mentally ill patients and their expectations towards psychiatric institutions. METHODS: The literature was selected from Medline covering the years 1996 - 2002. 342 articles were reviewed, 145 of which were described in this review. RESULTS: The burden of relatives are manifold and can be classified into the following categories: time spent on caring, financial difficulties, occupational restrictions, detrimental effects on relatives own physical and psychological well-being, reduction in leisure activities, negative effects on social relationships, experiences of discrimination and refusal, deficiencies in information about illness, feelings of not being taken seriously, insufficient service support, long distance to mental health service, emotional burdens of caregivers and difficulties with the patients behaviour. The expectations of the relatives mainly refer to the categories "relationship between staff and relatives", "information about illness" and "establishing of institutions required". CONCLUSION: The various burdens of relatives and their expectations towards psychiatric services point to necessary improvements of mental health services in the sense of a consumer-oriented psychiatric care. PMID- 12624849 TI - [Psychiatric emergency cases in Hamm/Westfalen]. AB - PURPOSE: Our study examines the frequency and further characteristics of psychiatric emergencies in the ambulance service of doctors on call in the city of Hamm. METHODS/PATIENTS: All emergency documentations of the year 2000 were retrospectively examined and evaluated with regard to psychiatric disorders according to the ICD-10. In Hamm (181, 197 inhabitants) there were 3812 emergency calls for doctors on call in the examined period. RESULTS: Due to internal emergencies, psychiatric diseases were diagnosed twice as often. Alcoholic intoxications (32.8 %), suicide attempts (16.8 %) and alcoholic withdrawal syndromes were mostly diagnosed in psychiatric emergencies. Most patients were male. The average age was 41.7 years. CONCLUSIONS: In the education for doctors on call, the training in the subject psychiatry is very low. Therefore the education and training of the diagnostic and therapy of psychiatric disorders must be intensified in the acquiring of the certification for ambulance service. acquiring PMID- 12624850 TI - [Life events and contact behaviour in drug-related death]. AB - The purpose of the study was to analyse influencing factors on drug-related death. Contact behaviour and life events in the forefield of drug-related death were of special interest. During a period of 5 years 189 drug-related deaths were investigated in the German town Essen (inhabitants about 600,000). 53.4 % of 189 people with drug-related death had contact to specialised institutes for drug addicts during lifetime. However, only about 15 % of this group contacted these institutes over the period of the last three months. Additionally, 26 of the 189 persons had a psychiatric comorbid diagnosis, and this subgroup also demonstrated a restricted contact behaviour. Life events during lifetime and especially during the last 3 months seemed to be of influence upon the circumstances of drug related death. According to this study, there seems to exist a turning away from professional institutions towards the end of the drug career which makes the documentation of influencing factors on drug-related death more difficult. Turning away from the help system is not the reason for drug-related death, but an important attendant circumstance. PMID- 12624851 TI - [Does acupuncture influence the cardiac autonomic nervous system in patients with minor depression or anxiety disorders?]. AB - We undertook this study to evaluate the effects of needle acupuncture on cardiac autonomic nervous system (ANS) function in patients with minor depression or anxiety disorder. Patients (n = 36) were randomly distributed into a verum acupuncture (VA) group (needles were applied at classical acupuncture points, e. g. He7, Pe6, Du20, Bl62, Ex6) or a placebo (PL) group (needles were applied only epidermal at non-acupuncture points). Both groups underwent standardized measurements of the 5-minute resting heart rate variability (HRV), which were performed before the first and after the 9th acupuncture session of an acupuncture series, and also three times (before the start and 5, respectively, 15 minutes after needle application) during the third acupuncture session. Demographic data between the VA and PL group did not differ. Before the start of acupuncture there were also no significant differences in HRV data between these groups. Compared to PL the VA group showed a significant decrease of the mean resting heart rate both, 5 and 15 minutes after needle application, combined with a trend towards an increase of the high frequency (HF; 0.15 - 0.4 Hz) and a decrease of the low frequency (LF; 0.04 - 0.15 Hz) spectral power. The latter effects resulted in an overall significant decrease of the mean LF/HF ratio in VA compared to PL treated patients. This pattern of findings suggests that in patients with minor depression or anxiety only verum acupuncture 1.) leads to a relative increase of cardiovagal modulation of heart rate and 2.) facilitates the physiological regulatory ANS function in response to alterations of external or internal environment. Clinical implications of these findings are discussed. PMID- 12624852 TI - [Homocysteine and dementia]. AB - Homocysteine is a vascular risk factor including cerebral macroangiopathy and microangiopathy. Furthermore, there might also be an association with cognitive disorders including vascular dementia and Alzheimer's disease. Hyperhomocysteinemia linked with cognitive impairment might be an indirect marker for low concentrations of vitamin B 12, vitamin B 6 or folate, resulting from low intake or from an impaired transport of the vitamins to the brain. Another possibility is a direct harmful effect of homocysteine to cognition via vascular and neurotoxic pathophysiologic mechanisms. Because hyperhomocysteinemia is a potentially reversible risk factor and can be identified early, it should be investigated by prospective intervention studies whether lowering homocysteine levels by vitamin supplementation could reduce incidence and progression of cognitive disorders. PMID- 12624853 TI - [Cognitive impairment in adults with neurofibromatosis type 1]. AB - Cognitive impairment is a common phenomenon in children with neurofibromatosis type 1 (NF1), but only little is known about its nature and frequency in adult NF1-patients. Using a comprehensive psychometric test battery, we investigated 20 patients with NF1 and 20 age and gender matched control subjects without neurological diseases. Results showed slightly lowered test scores in patients compared with controls but no specific intellectual impairment. On a computerized test of selective attention, the NF1-group had significant slower reaction times. Also, three out of four memory tests and a test of visuoconstructive abilities showed poorer test results in the NF1-patients. Executive functions however were not affected. The findings agreed well with the test profile in NF1-children and supported the idea of a continuum between childhood and adulthood. Observations are discussed in the context of studies investigating the association of cognitive deficits with either intracranial lesions or alterations in the neurofibromin expression. PMID- 12624854 TI - Analysis of GNAS1 and overlapping transcripts identifies the parental origin of mutations in patients with sporadic Albright hereditary osteodystrophy and reveals a model system in which to observe the effects of splicing mutations on translated and untranslated messenger RNA. AB - Albright hereditary osteodystrophy (AHO) is caused by heterozygous deactivating GNAS1 mutations. There is a parent-of-origin effect. Maternally derived mutations are usually associated with resistance to parathyroid hormone termed "pseudohypoparathyroidism type Ia." Paternally derived mutations are associated with AHO but usually normal hormone responsiveness, known as "pseudo pseudohypoparathyroidism." These observations can be explained by tissue-specific GNAS1 imprinting. Regulation of the genomic region that encompasses GNAS1 is complex. At least three upstream exons that splice to exon 2 of GNAS1 and that are imprinted have been reported. NESP55 is exclusively maternally expressed, whereas exon 1A and XL alphas are exclusively paternally expressed. We set out to identify the parental origin of GNAS1 mutations in patients with AHO by searching for their mutation in the overlapping transcripts. This information would be of value in patients with sporadic disease, for predicting their endocrine phenotype and planning follow-up. In doing so, we identified mutations that resulted in nonsense-mediated decay of the mutant Gs alpha transcript but that were detectable in NESP55 messenger RNA (mRNA), probably because they lie within its 3' untranslated region. Analysis of the NESP55 transcripts revealed the creation of a novel splice site in one patient and an unusual intronic mutation that caused retention of the intron in a further patient, neither of which could be detected by analysis of the Gs alpha complementary DNA. This cluster of overlapping transcripts represents a useful model system in which to analyze the effects that mutant sequence has on mRNA-in particular, splicing-and the mechanisms of nonsense-mediated mRNA decay. PMID- 12624855 TI - Is aspirin still the antiplatelet drug of choice for patients with peripheral arterial disease? PMID- 12624856 TI - Lessons learned...a tribute to a mentor. PMID- 12624857 TI - Differentiating quality improvement from research. AB - Monitoring of quality of care has always been an important part of health care. Self-regulation and external standards require care providers to furnish safe environments for the patient. Similarities in methods used by quality improvement (QI) projects and clinical research have created some confusion in differentiating the two practices. This article reviews the current literature and differences between QI and research. In addition, the article identifies and discusses four criteria: intervention, risk, audience, and data source, which allow investigators to differentiate between the two practices and follow the appropriate procedures for project review. PMID- 12624858 TI - Validation of the Hendrich II Fall Risk Model: a large concurrent case/control study of hospitalized patients. AB - This large case/control study of fall and non-fall patients, in an acute care tertiary facility, was designed to concurrently test the Hendrich Fall Risk Model. Cases and controls (355/780) were randomly enrolled and assessed for more than 600 risk factors (intrinsic/extrinsic). Standardized instruments were used for key physical attributes as well as clinician assessments. A risk factor model was developed through stepwise logistic regression. Two-way interactions among the risk factors were tested for significance. The best fitting model included 2 Log L chi square statistic as well as sensitivity and specificity values retrospectively. The result of the study is an easy to use validated Hendrich Fall Risk Model with eight assessment parameters for high-risk fall identification tested in acute care environments. PMID- 12624859 TI - Women's feedback on a chemopreventive trial for cervical dysplasia. AB - This study identifies the risks, benefits, and social influences affecting the decision to participate in a chemoprevention trial for cervical dysplasia. Women from three clinical sites in southeastern Michigan were invited to participate in a semi-structured interview. In previous studies, minority women have significant barriers to participation in clinical trials. However, in this study, women (63% black, 26% white, and 9% other ethnicity), were interested in participating and reported only moderate barriers. These barriers could easily be addressed with resources available to investigators. PMID- 12624860 TI - Comparing patient and nurse perceptions of perioperative care quality. AB - This study compared surgical patients' (n = 874) and perioperative nurses' (n = 143) perceptions of the quality of perioperative nursing care. The data were collected with a structured questionnaire in five hospital operating departments in Finland. The questionnaire items were divided into five main categories (staff characteristics, nursing activities, preconditions, progress of nursing process and environment); some of these categories were further divided into subcategories. Overall, patients tended to give significantly higher (P <.001) ratings than nurses, but for some items the patients had more critical perceptions. The results provide important clues for improving the quality of patient care so that staff activities better serve the needs of patients. PMID- 12624861 TI - The effectiveness of a self-care intervention for the management of postpartum fatigue. AB - The purpose of this study was to test the effectiveness of the Tiredness Management Guide (TMG) as a self-care intervention for the management of postpartum fatigue from the second to the sixth week postpartum. An experimental repeated measures analysis of variance design was used with a sample of 68 primiparous mothers. Interaction effects between group membership and time in the hypothesized direction were found for fatigue. When the data were tested from the second through the fourth week, a significant interaction effect was found between group membership and time with the experimental group having lower morning fatigue. Results suggest that using the TMG may reduce levels of morning postpartum fatigue from the second through the fourth week postpartum. PMID- 12624862 TI - Overcoming subject recruitment challenges: strategies for successful collaboration with novice research agencies. AB - In most studies, the investigator reaches out to locate potential research subjects using direct strategies such as targeted radio advertising, recruitment posters, and newspaper advertisements. However, other studies may depend on indirect methods of locating potential subjects and work through intermediary contacts in clinics and hospital outpatient departments. Some agency personnel may not have had prior experience with clinical research protocols and may be unfamiliar with screening potential subjects; these personnel are likely to be employed in agencies unaffiliated with academic health sciences centers. In cases in which agency staff members are new to clinical research, special attention is required to keep agency personnel updated about the status of the research and subject recruitment efforts. This article provides an overview of practical tips designed to engage and sustain the interest of novice research agencies in subject recruitment. The article concludes with a case overview of recruitment issues that occurred during a clinical trial addressing occupational low back pain. PMID- 12624863 TI - Identifying possible depression in clinical research: ethical and outcome considerations for the investigator/clinician. AB - Most nurse researchers are aware of the ethical implications of research including the need to ensure confidentiality for research participants, as well as to provide full disclosure of the risks and benefits of the study. However, another consideration that can impact methodology from the perspective of the protection of human subjects from research risks includes the measurement of certain psychosocial variables such as depression and appropriate disclosure of such information to research participants. This issue has received little attention in the nursing literature. The intent of this article is to provide an overview of (1) the importance of informing participants about possible depression, (2) issues of identifying potential depression in clinical research including those related to depression measures, (3) strategies for the investigator/clinician to ensure protection of research participants and potential impact of those strategies, and (4) considerations in addressing depression in study design and analysis. PMID- 12624864 TI - Academic nursing clinic: impact on health and cost outcomes for vulnerable populations. AB - The purpose of this study was to examine selected health and cost outcomes of clients who used an academic nursing clinic (ANC) located in a high-rise public housing facility for low-income citizens. Service use, health promotion and screening, quality of care, satisfaction and costs were examined. Health outcomes were improved. Estimated cost savings were about $36,000 during the first year with reduced paramedic and police calls, hospitalizations, and emergency room visits. Findings show that advanced practice nurses can positively influence health outcomes by providing cost-effective quality health care. PMID- 12624865 TI - Could lack of clarity in written advance directives contribute to their ineffectiveness? A study of the content of written advance directives. AB - Since 1991, advance directives have failed to have a significant impact on end of life care. One reason is that despite widespread interest in these documents, few individuals have chosen to issue advance directives. Another reason reported in several studies is that the conditions under which the patient intended the directives to apply are unclear as are some of the treatments requested or declined. The purpose of this study was to describe instructions commonly included in written advance directives concerning the conditions under which the directives should apply and the treatments that are requested or declined. Five hundred consecutive closed medical records were reviewed from the medical and surgical departments of an urban hospital. These records produced 40 (8%) advance directive documents. Patient instructions indicating when the directives should take effect and which treatments should be declined or provided were summarized. Nurses and other health professionals can use this information to assist in patients in making their advance directives as clear as possible and to make patients more aware of the potential limitations of these types of written instructions. PMID- 12624866 TI - How to choose and evaluate a research instrument. PMID- 12624867 TI - [Stress management in manufacturing industries]. AB - Job strain factors and stress management for workers in the manufacturing industries were reviewed in this article. Major job stress factors included high job demands, low job control, low social support, role ambiguity and conflict, the physical, chemical and ergonomics of the work environment, work patterns with work schedule and shift work, and job insecurity regarding future employment. In considering effective stress management plans which counteract the job stress factors mentioned above, it is essential to use an organizational approach in the work environment. For workers in the manufacturing industry, through this approach, it is important to promote more autonomy and activities with increasing job control, to give more clarified roles and responsibilities, to provide a more mutually supportive system with better ways to communicate and to introduce a system giving additional rewards, such as paid holidays for refreshment. Plans are also needed for individuals and groups, such as workers, managers, supervisors and workgroups, regarding education, skill training and mutual training according to the personnel characteristics of each target. In order to make these stress management plans successful in the long term, an actual system for stress management is required, with recognition and support by top management. It is also important to assess the effectiveness and method of each stress management plan with proper measurements. PMID- 12624868 TI - [The roles and duties of occupational health nurses in managing and coping with stress in the workplace]. AB - In recent years, employees who complain about too much stress have been increasing, and stressmanagement and coping with stress in the workplace have emerged as one of the important subjects in Occupational Health. There are two approaches to measures for stress. One is to decrease the stressors, and the other is to promote employees' tolerance of the stress reaction. Therefore the roles and duties of Occupational Health Nurses (OHNs) have been described for each approach. OHNs who are close to the employees as first line professionals support each employee to live healthly and independently, and aim at contributing to the employees' QOL. For that reason OHNs' roles in measures for stress must become much more important in the future. PMID- 12624869 TI - [Management of hyperuricemia in occupational health: with reference to "guidelines for the management of hyperuricemia and gout"]. AB - In 1996, the need for the clinical guidelines for the management of hyperuricemia and gout was proposed of the consensus conference held at the 29th annual meeting of the Japanese Society of Purine and Pyrimidine Metabolism (president Yuji Matsuzawa). At the consensus conference, the following announcement was made. 1. Because the majority of patients with hyperuricemia are in the condition of multiple risk factor clustering syndrome, hyperuricemia per se should be counted as one of the typical lifestyle related diseases. 2. Medical management should be directed independently for the treatment of gouty arthritis and for control of the serum uric acid level. 3. The serum uric acid level should be taken into account as a possible cardiovascular risk factor. 4. Urine alkalization should be started if there is no symptom indicating hyperuricemia for the prophylaxis of urinary stones and renal dysfunction. 5. All the medical management should be considered under the consensus of the many expert physicians dealing with hyperuricemia and gout. This principal announcement was made by the consensus conference and the simple management recommendation of a 6-7-8 rule was proposed through the consensus of expert physicians. Recently, a guideline committee was organized in the Japanese Society of Gout and Nucleic Acid Metabolism (previous by the Japanese Society of Purine and Pyrimidine Metabolism) and the Guidelines for the Management of Hyperuricemia and Gout were prereleased in February and published in August 2002. In the new guidelines, the above policy was introduced and evidence was collected to give the guidelines contemporary clinical usefulness and value. It will help in the proper management of hyperuricemia in apparently healthy persons in occupational health, having multiple risk factors. PMID- 12624871 TI - Evaluation of suspected immunodeficiency. PMID- 12624870 TI - [The impact of work and organizational characteristics on the health status, job dissatisfaction and turnover intentions of workers in an information service industry]. AB - Based on the conceptual model of "healthy work organization", we conducted a questionnaire survey of 612 Japanese workers in an information service industry company to investigate the effect of work stressors and organizational characteristics on workers' health status, job dissatisfaction and turnover intentions from July to August 2001. The response rate was 96.2%. For the statistical analysis, data on 488 computer technical support staff were used. To grasp the occupational stressors, we used a focus group to clarify work stressors and organizational characteristics. After factor analysis, we identified seven factors composed of 29 items and created seven scales of work and organizational characteristics. As scales of "organizational characteristics", "insufficient evaluation system", "undeveloped management system", and "career and future ambiguity" were used. The remaining scales, "poor coworker support", "poor supervisor support", "insufficient office amenities" and "high job demands and control", were used as scales of "work and workplace characteristics". The results of multiple regression analysis showed significant relevance of organizational characteristics to health status, job dissatisfaction and turnover intentions of workers. They supported "healthy work organizations" as useful conceptual tools for the study of organizational health. PMID- 12624872 TI - Talking with S. Wayne Kay. Interview by Celia Stevens. PMID- 12624873 TI - Is this hospital legally liable in an employee fender-bender? PMID- 12624874 TI - Urinary adulterants and drugs-of-abuse testing. PMID- 12624875 TI - Richard Fairfax of OSHA talks about the bloodborne pathogens standard. Interview by Dennis Ernst. PMID- 12624876 TI - Addressing the problem of controlling procrastination. PMID- 12624877 TI - Wet mount bacteria reporting. PMID- 12624878 TI - The front line of smallpox preparedness. PMID- 12624879 TI - Putting safety in the blueprint. Patient safety is the guiding force for a new hospital. AB - St. Joseph's Community Hospital of West Bend (Wis.) is building a new hospital using innovative architectural and design features to foster patient safety. Coupled with a nonpunitive safety culture, St. Joseph's may represent the wave of the future. PMID- 12624880 TI - The art of interviewing your next CEO. AB - If conducted properly, an interview is a highly effective tool for evaluating managers at any level of an organization. Broad, general questions, gently put, elicit sincere behavior from others, allowing them the freedom to respond as they wish. Their responses tell us what they typically focus on. Moreover, the interviewee is inclined to find us "safe" to talk to. This reduces their fear or discomfort, which should be any interviewer's goal. Probing often elicits the most spontaneous, hence truest, behavior revealing who this person is and how he or she will respond in a particular position or situation. Good probes, which should be used frequently in any conversation, also convey the message that we care about the interviewee and his or her views. Interviewing means focusing on others. It should be an integral part of the board's repertoire of skills. It is also an integral part of being a good manager. Few people are emotionally capable of implementing the principles described here. It is incumbent on the board, therefore, to select those members who can conduct in-depth interviews for a new CEO. PMID- 12624881 TI - What you can do: the trustee, patient safety, and JCAHO. AB - The Joint Commission has developed patient safety standards that hospital leaders, including trustees, are responsible for implementing, along with National Safety Goals and recommendations. PMID- 12624882 TI - Patient safety. BPOC technology: an answer to new JCAHO safety goals. PMID- 12624883 TI - Workforce. A difficult prescription to fill. Hospitals search for new solutions to relieve the pharmacist shortage. PMID- 12624884 TI - Corporate responsibility laws and not-for-profits: getting ahead of the curve. PMID- 12624885 TI - Management of bronchiolitis in infants and children. PMID- 12624886 TI - Allowing patients to find meaning where they can. PMID- 12624887 TI - A mediation/medical advisory panel model for resolving disputes about end-of-life care. PMID- 12624888 TI - Extramural ethics consultation: reflections [correction of relections] on the mediation/medical advisory panel model and a further proposal. PMID- 12624889 TI - Care coordination and utilization review: clinical case managers' perceptions of dual role obligations. PMID- 12624890 TI - Irene's story. PMID- 12624891 TI - Irene's view. An interview by Sigal Klipstein. PMID- 12624892 TI - Irene's physician's view. An interview by Sigal Klipstein. PMID- 12624893 TI - The social worker's view. An interview by Stephen O'Neill. PMID- 12624894 TI - An ethicist's view. An interview by Lachlan Forrow. PMID- 12624895 TI - New reproductive options and the incest taboo. PMID- 12624896 TI - Moving from moral judgment to ethical reasoning. PMID- 12624897 TI - Improving availability and reliability. PMID- 12624898 TI - Improvements in ionisation. PMID- 12624900 TI - Delivering shared support services. PMID- 12624901 TI - Aiming at greater efficiency. PMID- 12624902 TI - No one-size-fits-all solution. PMID- 12624903 TI - Engineering ethics under close scrutiny. PMID- 12624904 TI - Total internal reflection fluorescence microscopy in cell biology. PMID- 12624905 TI - Reflection interference contrast microscopy. PMID- 12624906 TI - Second harmonic imaging microscopy. PMID- 12624907 TI - Fluorescence correlation spectroscopy. PMID- 12624908 TI - Fluorescence correlation spectroscopy of GFP fusion proteins in living plant cells. PMID- 12624909 TI - Building and using optical traps to study properties of molecular motors. PMID- 12624910 TI - Optical-trap force transducer that operates by direct measurement of light momentum. PMID- 12624911 TI - Application of optical traps in vivo. PMID- 12624912 TI - Cytomechanics applications of optical sectioning microscopy. PMID- 12624913 TI - Measurements of cell-generated deformations on flexible substrata using correlation-based optical flow. AB - The optical flow algorithm presented here is a robust method that rapidly yields a high-density field of substrate displacement vectors based on two optical images. We found that one of the limiting factors, at least for inexperienced experimentalists, is the consistency of focusing or the drift in microscope focus. However, with properly collected images the standard error of the measurement was estimated to be on the order of +/- 0.10 pixels. Finally, although the discussion has been focused on the displacement of flexible substrata, a similar method should be applicable for detecting movements on other types of images, as long as the movement involves a certain degree of local coordination. PMID- 12624914 TI - Single-molecule imaging of rotation of F1-ATPase. PMID- 12624915 TI - Molecular motors and single-molecule enzymology. PMID- 12624916 TI - Visualization of single molecules of mRNA in situ. PMID- 12624918 TI - GFP-fusion proteins as fluorescent reporters to study organelle and cytoskeleton dynamics in chemotaxis and phagocytosis. PMID- 12624917 TI - Single ion channel imaging. PMID- 12624919 TI - Dynamic imaging of cell-substrate contacts. PMID- 12624920 TI - Imaging mitochondrial function in intact cells. PMID- 12624921 TI - Dynamic imaging of neuronal cytoskeleton. PMID- 12624922 TI - Imaging calcium dynamics in developing neurons. AB - Here we describe the techniques developed to image Ca2+ signals in motile nerve growth cones both in culture and in the developing Xenopus spinal cord. We have used these methods to identify two spatially and temporally distinct classes of Ca2+ transients in growth cones. Imaging Ca2+ in morphologically complex migratory cells allows for analysis and correlation of discrete signals with a wide variety of cellular behaviors. For example, we find that localized Ca2+ changes at the tips of individual filopodia correlate with reduced filopodial motility. Further, rapid fixation after Ca2+ imaging made it possible to determine that transients occur at integrin receptor clusters that may generate and in turn be regulated by these local signals. We describe the use of caged Ca2+ to locally impose Ca2+ transients in individual filopodia and find this treatment sufficient to repel neurite outgrowth. Calcium signals across broad spatial and temporal dimensions are universal regulators of numerous complex and varied cellular functions. The imaging methods we describe here begin to view growth cones over a range of spatial resolutions and temporal frequencies necessary to detect different types of Ca2+ transients, however it is clear that not all dimensions have been examined. In particular, imaging cells more rapidly and at higher magnification may one day allow us to detect more elemental events such as single-channel openings, as has been achieved in nonneuronal cells. We also describe techniques used to examine Ca2+ signals in growth cones migrating within the spinal cord. These types of studies are ultimately necessary to confirm the relevance of in vitro findings. Although designed for the Xenopus spinal cord, the methods we outline should be applicable to other tissues and organisms. Finally, we use caged Ca2+ as a tool to reproduce very precise changes in cytosolic Ca2+ levels. This is a powerful means to test the function of different types of Ca2+ transients and assess the downstream regulators of those signals. These types of manipulations can also be used with other types of caged compounds, many of which are commercially available (Molecular Probes) or readily synthesized. PMID- 12624923 TI - Optical monitoring of neural activity using voltage-sensitive dyes. PMID- 12624924 TI - Steady-state fluorescence imaging of neoplasia. PMID- 12624925 TI - In vivo and ex vivo tissue applications of two-photon microscopy. PMID- 12624926 TI - Fluorescence anisotropy in pharmacologic screening. PMID- 12624927 TI - Surface and printing effects on fluorescent images of immobilized biomolecule arrays. PMID- 12624928 TI - [Phylogeny of Triticum L. and Aegilops L. genuses inferred from a comparative analysis of nucleotide sequences in promoter rDNA regions of individual species]. AB - The process of accumulation of knowledge on wheat and related wild species during the 20th century is briefly reviewed with special reference to the evidence of the recent years on evolution of polyploid wheats and the role of diploid species. The latter serve as potential donors of the genomes, detection of which is particularly important because of the continuing speciation in the tribe Triticeae and artificial development of synthetic forms. The arguments in favor of the donor role for various diploid wheat species and aegilopses from the section Sitopsis are compared. It is stated that in the formation of the both lines of polyploid wheats turgidum-aestivum and timopheevi, diploid Aegilops speltoides acted as a maternal form. In addition to plasmatic genomes, this aegilops species introduced into them also the B and G nuclear subgenomes. A comparison of nucleotide sequences in the variable part of the promoter of evolutionary conserved rRNA genes in polyploid wheats with their counterparts in diploid wheats and aegilopses confirmed the accepted wheat phylogenies. PMID- 12624929 TI - [Genetic analysis of spontaneous suppressors of the pho85 mutation in the yeast Saccharomyces cerevisiae]. AB - Chaperones are known to play an important role in complexation of cyclin dependent kinases with cyclins. In yeast cells growing in the presence of phosphate, cyclin-dependent kinase Pho85p and cyclin Pho80p form a complex and phosphorylate activator Pho4p. As a result, Pho4p is exported from the nucleus, and the PHO5 gene is not transcribed. The mutations suppressing the pho85 mutation were analyzed in order to identify genes which code for chaperones involved in the formation of the Pho80p-Pho85p complex in the presence of environmental phosphate. Dominant mutations DSP1, DSP2, and DSP4-6 were found. It is shown that the DSP1 gene is 2.1 cM away from the PHO85 gene on chromosome XVI and probably coincides with the EGD1 gene coding for a chaperone. PMID- 12624930 TI - [On the involvement of the regulatory gene prqR in the development of resistance to methyl viologen in cyanobacterium Synechocystis sp. PCC6803]. AB - The role of the prqR gene in the regulation of the adaptive response of the cyanobacterium Synechocystis sp. PCC6803 to the oxidative stress induced with methyl viologen (MV) was studied. For this, transcription activity of prqR and the genes, which may be involved in the control of resistance to MV, was determined by means of Northern blot hybridization in wild-type cells and in the MV-resistant Prq20 mutant with a mutation located in the DNA-binding domain of the PrqR protein. It was ascertained that the prqR gene is a component of the prqR-prqA operon and down regulates its transcription. In cells of the wild-type strain containing MV, the autorepressor activity of the PrqR protein enhances and transcription of mvrA and sodB genes encoding an respectively assumed transporter protein and iron-containing superoxide dismutase increases. The prqR gene may be involved in the negative, indirect control of transcription of these genes. The Prq20 mutant is characterized by an MV-independent derepression of the prqR-prqA operon and by a slightly increased transcription of mvrA and sodB genes not stimulated by MV. Nevertheless, the expression of mvrA and sodB genes was lower than in wild-type cells after the MV treatment. On the strength of this evidence, it is assumed that the main mechanism underlying for the resistance to MV in the Prq20 mutant is derepression of the prqA gene, the product of which is homologous to multidrug transporters, drug efflux proteins. PMID- 12624931 TI - [Heat shock during the development of brain structures of Drosophila: the memory development in the l(1)ts403 mutant of Drosophila melanogaster]. AB - The structures and functions of many genes are homologous in Drosophila and humans. Therefore, studying pathological processes in Drosophila, in particular neurogenerative processes accompanied by progressive memory loss, helps to understand the ethiology of corresponding human disorders and to develop therapeutic strategies. It is believed that the development of neurogenerative diseases might result from alterations in the functioning of the heat shock/chaperone machinery. In view of this, we used Drosophila mutant l(1)ts403 with defective synthesis of heat shock proteins for studying learning and memory in a test of conditioned courtship suppression following a heat shock given at different developmental stages. High learning indices were registered immediately and 30 min after training both in the intact controls and in flies subjected to different developmental heat shocks. This indicated normal learning and memory acquisition in the mutant. At the same time, memory retention (3 h after training) suffered to different extent depending on the developmental stage. The remote effects of heat shock given during the formation of the mushroom bodies indicated the important role of this brain structure in the memory formation. The observed memory defects may result from alterations both in mRNA transport and in the functions of molecular chaperones in the l(1)ts403 mutant. PMID- 12624932 TI - [Cytogenetic features of the blood-sucking blackfly Wilhelmia paraequina Puri (Diptera: Simuliidae) from Armenia]. AB - Three blackfly Wilhelmia paraequina populations of Armenia (rivers Debet and Megriget, channel Megri) were studied. 2n = 6: IS + IIL, IIS + IL, IIIS + IIIL. Cytological maps of polytene chromosomes were constructed. High inversion polymorphism (95.63%) was observed, with 2.6 inversions per individual. Three inversions proved to be associated with male development: Y1 was characterized by a combination of two heterozygous inversions, IIL-3 + IIL-1, while Y2 had another combination of heterozygous inversions, IIL-5 + IIL-3. The X chromosome had a standard homozygous IIL disk sequence. The populations were shown to be similar in autosomal polymorphism. A tendency for differentiation was observed with respect to frequencies and types of sex-linked inversions: the sex determination system was Y1X-XX in the Debet and Megriget populations and Y2X-XX in the channel (Megri) population. On the strength of these findings, one W. paraequina morphotype was assumed to involve two cytotypes, A (Debet and Megriget populations) and B (Megri population). PMID- 12624933 TI - [Analysis of transgenic tobacco plants carrying the gene for the surface antigen of the hepatitis B virus]. AB - The plasmids carrying the gene encoding the hepatitis B surface antigen (HBsAg) under the control of 35S RNA single or dual promoters of the cauliflower mosaic virus CaMV 35S were constructed. These constructions were used for obtaining transgenic tobacco plants that synthesize the HBS antigen. The presence of HBsAg in tobacco plant extracts was confirmed by the enzyme-linked immunoassay using antibodies against the native HBs antigen. The antigen amount in plants carrying the HbsAg gene under a single 35 S promoter was 0.0001-0.001 of the total soluble protein whereas the use of a dual 35S promoter increased the antigen synthesis to 0.002-0.05% of the protein. The antigen-synthesizing ability was inherited by the offspring. In the F1 plants, the antigen expression varied in different lines comprising 0.001 to 0.03% of the total soluble protein, which corresponded to the antigen amount in the F0 plants. PMID- 12624934 TI - [Genetic relationships among Far Eastern species of the family Araliacea inferred by RAPD analysis]. AB - A molecular genetic study of Far Eastern species of the family Araliaceae by means of RAPD analysis was conducted. Using 21 primers we assessed variability at 595 loci. Based on matrices of genetic distances D, dendrograms of genetic relationships among eleven species of six genera of this family were constructed. Our results suggest that Acanthopanax sessiliflorus and Eleutherococcus senticosus belong to different genera, Aralia cordata and A. continentalis are different species, and A. elata and A. mandshurica probably cannot be regarded as distinct species. Genetic similarity of Far Eastern A. cordata and American A. hispida is shown. PMID- 12624935 TI - [Analysis of the effects of parental genotypes of rye lines on the development of quantitative traits in primary octaploid triticale. Plant height]. AB - When breeding the primary spring octoploid triticale derived from crosses of various inbred rye lines to wheat Chinese Spring, the effects of the rye genotype and growth conditions on the plant height and proportion of the first, second, and final (pedicle) internodes to the entire stem length were studied. Two triticale groups were examined: homozygotes for the dominant (Ddw1) and recessive (ddw1) alleles of the gene responsible for short stem in rye. In the short stem triticale lines carrying the Ddw1 alleles, the plants were 20 cm shorter on average than those in the ddw1-carrying lines, and the distribution of the two triticale groups overlapped significantly. In both groups, the lines significantly differing in plant height could be differentiated, because of allelic diversity of the additional genes controlling this trait along with the Ddw gene. In most triticale lines, especially in the Ddw1-carrying ones, the plant height was much reduced under unfavorable growth conditions. At the same time, a short-stem line was isolated, which is characterized by ecological plasticity, like the maternal wheat cultivar. In the triticale studied, the stem structure depended on the short-stem rye genotype. The two-year study showed that in the triticale carrying the dominant allele of this gene, the first internode is significantly extended, whereas the upper (pedicle) internode is reduced, which increases plant lodging resistance. The differences revealed between the rye lines as well as their effect on the quantitative triticale traits are discussed in view of a variant of the hybridological analysis, which had been previously proposed for identification and mapping of the correspondent rye genes. PMID- 12624936 TI - [Phenotypic expression of the mutant gene diabetes insipidus in rats and criteria of genotyping by phenotype]. AB - The autosomal semidominant mutant gene di (diabetes insipidus) is manifested in homozygotes in the form of diabetes insipidus with water consumption from 25 to 100% of body weight per day. The heterozygotes di/+ drink water at a rate higher than 5% but lower than 25%. The level of water consumption in rats with +/+ genotype does not exceed 5% of body weight per day. Segregation analysis of F1 animals yielded by various crosses showed that genotyping of di/di homozygotes is absolutely reliable at 30% and higher level of the water consumption per day. PMID- 12624937 TI - [Phylogenic and geographic analysis of chum salmon Oncorhynchus keta (Walbaum) in Asian populations based on mitochondrial DNA variation]. AB - We used restriction length polymorphism (RFLP) analysis of PCR-amplified fragments of mtDNA to study the genetic structure of chum salmon populations sampled in 1993-2000 during a spawning run in five rivers: Narva (Southern Primorye), Naiba (Sakhalin Island), Sernovodnaya (Kunashir Island, Southern Kuril Islands), Ola (northwestern coast of the Sea of Okhotsk), and Anadyr' (Chukotka Peninsula). In total, 49 haplotypes were identified in 193 fish. Heterogeneity tests showed highly significant (P = 0) differences among all sample pairs. The estimated time of independent divergence of the populations or population groups is in good agreement with the time of Pleistocene glaciations. This result suggests that it is cyclic global changes during this time period that were crucial in determining the within-species divergence in chum salmon. The types of mtDNA genetic variability and mismatch distribution between haplotypes in the populations indicate that the southern regions of the Sea of Okhotsk and Sea of Japan served as refugia for chum salmon during glaciation periods. PMID- 12624938 TI - [Genetic subdivision of Dagestan ethnic populations]. AB - Relationships between ethnic and genetic differentiation with respect to 54 microsatellites have been analyzed in five Daghestan ethnic groups. To detect the microsatellites, human chromosomes 3, 17, and 18 were screened with a step of 10 cM (Weber/CHLC 9.0 markers) at the Mammalian Genotyping Service (National Institute of Health, United States). Comparison of the polymorphism of these loci in Daghestan populations with average worldwide data has revealed generally low heterozygosity in Daghestan populations, which is accounted for by traditional endogamous and consanguineous marriages throughout the history of these populations. The inbreeding coefficient in Daghestan ethnic groups varies from 0.005 to 0.0134 and is close to the worldwide maximum known to date. For some DNA loci, significant differences between the offsprings of consanguineous and exogamous marriages with respect to allele sizes and their variance have been found. The Daghestan ethnic populations studied differ from one another in both the frequencies of common alleles and the presence of rare alleles that are unique for each ethnic group of Daghestan and have not been found in any other population in the world. PMID- 12624939 TI - [Genetic determination of mental activity parameters in the families of schizophrenic patients]. AB - Psychological parameters of mental activity (30 in total) and their genetic determination were studied in 67 families of schizophrenia patients (67 patients, 107 parent, and 30 sibs). Abnormalities of most of the examined characteristics were found in both the patients and their healthy relatives. Parameters of attention shifting and emotionality exhibited the largest genetic component (25 75 and 17-98%, respectively) in all analyzed groups of relatives (probands affected sibs, probands-healthy sibs, healthy parents-healthy children, affected parents-affected children). Significant impact of genetic factors was also found in parameters "steadiness of attention under conditions of continuous concentration," "mediated retention span" "productivity of arbitrary retention by reproduction data," "personal anxiety level," "reflection of unusual social groups," and "self-assessment." The relationships among the characteristics examined in the system of mental activity were established by means of cluster analysis. The results of this study can be used in medical genetic counseling for identifying persons at high risk for schizophrenia. PMID- 12624940 TI - [Comparative restriction enzyme analysis of mitochondrial DNA in several populations of lake resident chars of the genus Salvelinus]. AB - Restriction enzyme analysis was employed in studying the mitochondrial DNA (mtDNA) ATPase6/ND4L region in several Northeast Asian populations of resident lake char of the genus Salvelinus. On evidence of mitotypes, genetic similarity was assumed for populations of neiva (Ueginskoe Lake), lake resident char from the northern coast of the Sea of Okhotsk (Mak-Mak Lake and Elekchanskie lakes), and dolly varden. Mitotype AAAA proved to be common for these populations. Lake char populations of the Juliet and Maxi lakes (the basin of the Kolyma river) had mitotype DBAA, which is similar to mitotype DBAB observed earlier in Taranetz char S. taranetzi from Chukotka. The divergence between mitotypes AAA and DBAA was estimated at 0.3%. Different origins were assumed for the lake resident char populations from the basins of the Sea of Okhotsk and of the Kolyma River, the former originating in the Pacific and the latter, in the Arctic basins. PMID- 12624941 TI - [Genetic and demographic structure of Russian populations from the Tver' and Rostov regions]. AB - Based on the data from 569 questionnaires collected in Udomlya and Ostashkov raions of Tver oblast and 436 questionnaires collected in Tsimlyansk and Dubovsk raions of Rostov oblast, genetic demographic characteristics and Crow's index for urban and rural populations of these regions were calculated. The data obtained were compared with those for other Russian populations obtained earlier. PMID- 12624942 TI - [Differentiation antigens in tumors: dependence on carcinogenesis mechanisms and progression (a hypothesis)]. AB - The data considered in the paper indicate that a tumor clone resulting from cell transformation, in order to develop, should overcome a microenvironmental constraint. This destroys intercellular contacts and cell interactions with extracellular matrix required for induction and maintenance of epithelium differentiation. The possible reasons for this lie in mutations of genes that control cell adhesion molecules and integrins, as well as proteases secreted by a tumor. These events lead to partial loss of differentiation antigens by a cell or to their incorrect localization in a cell. Simultaneously, the expression of embryo-specific genes is unblocked, leading to overexpression of embryonic antigens and their abnormal secretion into blood, which results in appearance of oncofetal markers in blood. Discussed from this point of view are alpha fetoprotein, the carcinoembryonic antigen, and the prostate-specific antigen, which are used as tumor markers. PMID- 12624943 TI - [Functional interactions of the influenza virus glycoproteins]. AB - Hemagglutinin (HA) and neuraminidase (NA) are functionally related coat glycoproteins of the influenza virus (Flu). HA interacts with terminal sialyl residues of oligosaccharides and ensures the binding of the virus particle to the cell surface. NA is a receptor-destroying enzyme that removes sialyl residues from oligosaccharides contained in cell and virus components and thereby prevents aggregation of virus particles. Analysis of reasortants combining low-functional NA of human Flu with HA of avian Flu showed that sialyl residues are not completely removed in some cases. With high HA affinity for sialyl substrates, such virus particles aggregate, aggregates accumulate on the cell surface, and virus yield decreases. Serial passaging of low-yield aggregating reassortants may lead to selection of high-yield variants, which do not aggregate. A loss of aggregation is due to a decrease in HA affinity for high-molecular-weight sialyl substrates. On evidence of sequencing of the HA gene in original reassortants and their nonaggregating variants, HA affinity is reduced and aggregation lost owing to a mechanism common for different HA antigenic subtypes (H2, H3, H4, and H13). This is an increase in the negative charge as a result of an amino acid substitution in the vicinity of the receptor-binding pocket of HA. Taken together, these findings suggest a way of postreassortment adaptation which improves the functional match of HA and NA. The experimental system employed provides a model of natural processes associated with generation of Flu variants having a pandemic potential. PMID- 12624944 TI - [Immunomodulatory proteins of orthopoxviruses]. AB - The review considers recent data on the structural-functional organization of the genome of orthopoxviruses pathogenic for humans, including the variola, monkeypox, cowpox, and vaccinia viruses. Emphasis was placed on the structure of molecular virulence factors that suppress the inflammatory reactions, immune response, and interferon effects induced by virus infection. PMID- 12624945 TI - [Association of the G7831A polymorphism of the ACE gene with myocardial ischemia in the Moscow population]. PMID- 12624946 TI - [Association of the T174M and M235T polymorphisms of the angiotensinogen gene with myocardial ischemia in the Russian population of the city of Moscow]. PMID- 12624947 TI - [The mutation spectrum of the CFTR gene in mucoviscidosis patients from Bashkortostan]. AB - Mutations of CFTR were studied in patients with cystic fibrosis (CF) from Bashkortostan. In total, 15 mutations were observed and 51% of all mutant alleles identified. The most diagnostically significant mutations were delF508 (33.8%), 394delTT (3.52%), CFTRdele2.3(21 kb) (1.41%), R334W (1.41%), 3849+ 10 kbC-->T (1.41%), and N1303K (1.41%). Mutations G542X, 2184insA, S1196X, and W1282X were each found in less than 1% patients. Five new mutations and two neutral substitutions were revealed. These were I488M (exon 10), 1811 + 12A-->C (intron 11), T663S (exon 13), I1226R (exon 19), 4005 + 9A-->C (intron 20), 2097A-->C (A655A, exon 13), and 3996G-->C (V1288V, exon 20). Bashkortostan was shown to differ in CFTR mutation spectrum from other regions of Russia. The results will allow direct DNA diagnostics of CF in far more families. Molecular screening of probands' relatives will contribute to identification and medical genetic counseling of heterozygous carriers, which is essential for CF prevention. PMID- 12624948 TI - [Polymorphism of the serotonin receptor (5-HTR2A) gene and verbal fluency in normalcy and schizophrenia]. AB - To study the effect of the serotonergic brain system on verbal fluency (i.e., the ability to rapidly extract necessary words from the vocabulary), the T102C polymorphism of the serotonin receptor type 2A (5-HTR2A) gene was tested for association with verbal fluency in 108 patients with schizophrenia or schizotypic disorders and 97 mentally healthy individuals. A significant association was observed only in male schizophrenics (N = 67), with homozygotes A2A2 having lower verbal fluency. The results did not support the association between the 5-HTR2A polymorphism and verbal fluency in normalcy, and agree with the assumed contribution of genotype A2A2 to the severity of schizophrenia. PMID- 12624949 TI - [Molecular genetic analysis of Y-chromosome micro deletions in men with severe spermatogenic defects]. AB - Microdeletions of the Y-chromosomal AZF loci were revealed in 10 (12%) of 82 patients with severe idiopathic spermatogenetic defects. Deletions involved AZFc in six patients, AZFa in one patient, AZFb + c in two patients, and AZFa + b + c in one patient. Microdeletion analysis employed multiplex PCR with 22 pairs of primers directed to Y-specific STS of deletion intervals 5, 6, and 7 (Yq11). Spermatogenesis in men with AZF microdeletions was assessed with semen analysis, microscopic examination of testicular aspirate, and quantitative karyotypic analysis of immature germline cells in ejaculate or aspirate. The character of spermatogenetic defects was correlated with the size and location of microdeletions in order to study the genotype-phenotype relationship. PMID- 12624950 TI - [A novel approach to isolation and functional characterization of genomic DNA from the methylotrophic yeast Hansenula polymorpha]. AB - A novel approach to isolation and functional characterization of the Hansenula polymorpha genes basing on the use of two strains of different origin is described. One of these strains is better suited for the isolation of genomic DNA fragments, while the other is preferable for their functional analysis. Thirty three genomic sequences governing expression of a reporter protein have been isolated. Analysis of the sequence encoding a homolog of the Saccharomyces cerevisiae cofilin revealed two introns. Another isolated DNA fragment encoded a homolog of the S. cerevisiae V ps10p. Disruption of the corresponding gene resulted in secretion of a vacuolar protein, carboxypeptidase Y, into the culture medium. PMID- 12624951 TI - [On parametric stability of gene networks controlling ontogenetic processes]. AB - The problem of evaluating the parametric stability of three models of pro- and eukaryotic gene networks controlling ontogenetic processes has been defined and solved. Experimental plans of testing gene networks for parametric stability based on the method of generalized threshold models were developed and realized as a software application. We examined the "sensitivity" of the functioning modes to random variations of the parameters in the three model systems: the system of developmental control of phage lambda, the subsystem of morphogenetic control of Arabidopsis thaliana flower, and the gene subnetwork controlling early ontogeny in Drosophila melanogaster. The parametric stability was quantitatively assessed for these models. PMID- 12624952 TI - [Gene expression profiles of protein kinases and phosphatases obtained by hybridization with cDNA arrays: molecular portrait of human prostate carcinoma]. AB - Hybridization with cDNA arrays was used to obtain expression profiles of 263 protein-tyrosine kinase (PTK), protein-tyrosine phosphatase (PTP), dual-specific phosphatase (DuSP), and other genes for the normal prostate tissue, primary prostate carcinomas (PC) of 84 patients, 7 xenografts, and 5 carcinoma cell lines. Analysis of 96 profiles revealed eight clusters of genes coexpressed in PC (coefficient of correlation r > 0.7). According to the known functions of their genes, the clusters were designated as proliferating-cell (CDC42, TOP2A, FGFR3, MYC, etc.), neoangiogenesis and blood-cell (LCK, VAV1, KDR, VEGF, MMP9, SYK, PTPRS, and FLT4), invasion-1 and invasion-2 (ADAM17, TRPM2, DUSP6, VIM, CAV1, CAV2, JAK1, PTPNS1, FYN, and PDGFB), HER2, and PSA/PSM/HER3. Basing on expression profiles of 66 genes, a molecular classification of PC was constructed and allowed discrimination between PC and cell lines or xenografts at 98.9% probability. The results suggested that, along with PSA, PSM (FOLH1), kallikrein 2, and a-2-macroglobulin, cell signaling genes EGFR, HER2, HER3, TOP2, KRT8, KRT18, VEGF, CD44, VIM, CAV1, and CAV2 may serve as diagnostic and prognostic markers in PC. The HER2, VEGF, and CD44 genes and the MMP and ADAM families were assumed to be promising targets for inhibitors of PC cell proliferation and metastasis. PMID- 12624953 TI - [Dominant-negative inactivation of p53: the effect of the proportion between trans-dominant inhibitor and its target]. AB - Dominant-negative mutations of the p53 tumor suppressor gene and oligomerization of the mutant and wild-type p53 are considered responsible for functional inactivation of the p53 tetramer. Although dominant-negative inactivation of p53 is well reproducible in experimental systems, its contribution to processes occurring in tumor cells heterozygous at p53 is still unclear. To study the effect of dominant-negative inhibitor GSE22 on the p53 activity, cultures coexpressing GSE22 and tetracycline-suppressible p53 were derived from p53 negative cell lines. Transcriptional activity and expression of p53 proved to depend on the proportion between p53 and GSE22. The dominant-negative effect was observed only when GSE22 was in a multifold excess to p53. GSE22 was shown to be suitable for complete reversible inactivation of p53. PMID- 12624954 TI - [The effect of modification of nucleotide-37 on the interaction of aminoacyl-tRNA with the A-site of the 70S ribosome]. AB - To estimate the effect of modified nucleotide-37, the interaction of two yeast aminoacyl-tRNAs (Phe-tRNAK+YPhe and Phe-tRNAK-YPhe) with the A site of complex [70S.poly(U).deacylated tRNA(Phe) in the P site] was assayed at 0-20 degrees C. As comparisons with native Phe-tRNAK+YPhe showed, removal of the Y base decreased the association constant of Phe-tRNAK-YPhe and the complex by an order of magnitude at any temperature, and increased the enthalpy of their interaction by 23 kJ/mol. When the Y base was present in the anticodon loop of deacylated tRNA(Phe) bound to the P site of the 70S ribosome, twice higher affinity for the A site was observed for Phe-tRNAK-YPhe but not for Phe-tRNAK+YPhe. Thus, the modified nucleotide 3' of the Phe-tRNA(Phe) anticodon stabilized the codon anticodon interaction both in the A and in the P sites of the 70S ribosome. PMID- 12624955 TI - [DNA-[N4-cytosine]-methyltransferase from Bacillus amyloliquefaciens: mechanism of action derived from steady state kinetics]. AB - Kinetic analysis of methyl group transfer from S-adenosyl-L-methionine (SAM) to the 5'-GGATCC recognition site catalyzed by the DNA-[N4-cytosine] methyltransferase from Bacillus amyloliquefaciens [EC 2.1.1.113] has shown that the dependence of the rate of methylation of the 20-meric substrate duplex on SAM and DNA concentration are normally hyperbolic, and the maximal rate is attained upon enzyme saturation with both substrates. No substrate inhibition is observed even at concentrations many times higher than the Km values (0.107 microM for DNA and 1.45 microM for SAM), which means that no nonreactive enzyme-substrate complexes are formed during the reaction. The overall pattern of product inhibition corresponds to an ordered steady-state mechanism following the sequence SAM decreases DNA decreases metDNA increases SAH increases (S-adenosyl-L homocysteine). However, more detailed numerical analysis of the aggregate experimental data admits an alternative order of substrate binding, DNA decreases SAM decreases, though this route is an order of magnitude slower. PMID- 12624956 TI - [Spliced oct-1 gene mRNA isoforms with untranslated exons and a deletion in the region coding for the POU-specific domain]. AB - Transcription factor Oct-1 is involved in expression regulation of housekeeping genes, in lymphocyte differentiation, and in the immune response. Tissue-specific oct-1 mRNA isoforms are known to be expressed in lymphoid cells. Four new mouse isoforms were identified. Of these, two were tissue-specific (oct-1R alpha and oct-1R beta) and contained exon 1L. The oct-1R alpha was shown to contain an additional fragment, which corresponds to an exon located in the 3'-region of mouse otf-1. No homolog was found in human OTF-1. The oct-1R alpha isoform proved to lack an exon coding for a fragment of the POU domain. This deletion results in a loss of the first helix of the domain, and the mutant protein is devoid of affinity for octamer ATGCAAAT. Two other mRNA isoforms, oct-1d and oct-1e, were shown to contain untranslated regions between exons 1U and 2. The regions correspond to exons 1i and 2i located between exons 1U and 1L in the 5'-region of the mouse oct-1 gene. Human OTF-1 was not found to contain exon 1i. On evidence of these and published data, it was assumed that a set of oct-1 isoforms is present in the cell, reflecting the complexity of expression regulation of oct-1 and the multiplicity of its functions. PMID- 12624957 TI - [The mRNA codon environment at the P and E sites of human ribosomes deduced from photo crosslinking with pUUUGUU]. AB - Three mRNA analogs--derivatives of hexaribonucleotide pUUUGUU comprising phenylalanine and valine codons with a perfluoroarylazido group attached to the C5 atom of the uridine residue at the first, second, or third position--were used for photocrosslinking with 80S ribosomes from human placenta. The mRNA analogs were positioned on the ribosome with tRNA recognizing these codons: UUU was at the P site if tRNA(Phe) was used, while tRNA(Val) was used to put there the GUU codon (UUU at the E site). Thus, the crosslinking group of mRNA analog might occupy positions -3 to +3 with respect to the first nucleotide of the codon at the P site. Irradiation of the complexes with soft UV light (lambda > 280 nm) resulted in the crosslinking of pUUUGUU derivatives with 18S RNA and proteins in the ribosome small subunit. The crosslinking with rRNA was observed only in the presence of tRNA. The photoactivatable group in positions -1 to +3 binds to G1207, while that in positions -2 or -3 binds to G961 of 18S RNA. In all cases, we observed crosslinking with S2 and S3 proteins irrespective of the presence of tRNA in the complex. Crosslinking with S23 and S26 proteins was observed mainly in the presence of tRNA when modified nucleotide occupied the +1 position (for both proteins) or the -3 position (for S26 protein). The crosslinking with S5/S7 proteins was substantial when modified nucleotide was in the -3 position, this crosslinking was not observed in the absence of tRNA. PMID- 12624958 TI - [Use of the modified method of chromatin immunoprecipitation for the isolation of actively transcribed loci]. AB - A modified version of the chromosomal immunoprecipitation (ChIP) assay was implemented for discrete isolation and characterization of actively transcribed genes. Specifically, it was demonstrated with the gene II/9-1 of Sciara coprophila as a model locus that significant enhancement in the isolation of actively transcribed versus repressed and inactive genes can be achieved through the ChIP methodology. A combination of solid-phase magnetic bead technology with chromosomal immunoprecipitation using antibodies that recognize the large subunit (c) of RNA polymerase II resulted in efficient isolation of the promoter region of gene II/9-1 exclusively during the amplification stage of larval development, when the gene is actively transcribed. It is postulated that the novel technology described herein can be applied to a wide variety of systems for efficient isolation and in vivo assessment of actively transcribed genes regulated by virtually any given transcription factor. PMID- 12624959 TI - [What forces can determine the formation of highly specific protein-protein complexes?]. AB - A software package was designed and used in a detailed study of the contact regions (interfaces) of a large number of protein-protein complexes using the PDB data. It appeared that for about 75% of the complexes the amino acid composition of the subunit surface in the contact region is not essential. Thus one may suggest that, along with the amino acid residues at the interface, the residues in the interior of the globules substantially contribute to protein-protein recognition. Such interactions between quite remote residues are most probably of electrical nature, and are involved in recognition by contributing to the overall electric field created by the protein molecule; the configuration of this field is perhaps the definitive factor of recognition. The overall field of the protein molecule is additively built of the fields created by each constituent residue, and it can be calculated as a sum of the fields created by the protein multipole (aggregate of 'partial' electric charges assigned to every atom of the protein molecule). Preliminary calculations of the remote electrostatic interaction have been performed for ribonuclease subunits in vacuum. The results are indicative of a real possibility that the electric field created by the protein multipole can strongly influence the mutual orientation of molecules before Brownian collisions. PMID- 12624960 TI - [Structure and seasonal dynamics of the biotic community ectoparasites of the bank vole in the Il'men'-Volkhov lowland]. AB - A biodiversity, seasonal dynamics and parasite load of a single individual and local population of the bank vole (Clethrionomys glareolus) were studied in coniferous and mixed forests of the Ilmen-Volkhov lowland (neighborhood of Oskuy village, Chudovo region, Novgorod Province) in the period from June 1999 till May 2002. The Gero's traps were used for collecting the host. Lines of traps stood in each place during 3-5 days and were checked twice a day. Trapping of micromammalian hosts and collecting of parasites took place each month, except a few gaps. Total number of collected animals 2854 including 1405 bank voles. The 29 ectoparasite species were recorded on the bank vole in the area of study. Among some mites and fleas a few species are accidental parasites probably accepted by voles from other species of animals. In the Oskuy area, the bank voles are the main and additional hosts of 25 ectoparasite species: fleas--8, lice--1, ixodid ticks--2, gamasid mites--7, acariform mites 7. Species composition of ectoparasites, their occurrence and abundance change during the year. Seasonal changes of abundance and occurrence indices are most expressed in the temporary ectoparasites (ixodid ticks, chiggers, gamasid mites), while in the permanent parasites (lice, acariform mites: Myobiidae, Myocoptidae, Listrophoridae), the seasonal fluctuations of indices are displayed in a less scale. Most vole specimens were infected with this or that ectoparasite species. The parasitocenosis on an individual specimen usually included less than 10 species of the total number 29 species recorded on the bank vole in the area investigated. One ectoparasite species was recorded on 21% of host specimens, 2-5 ectoparasite species were found on 71% of host individuals. Maximal number of ectoparasite species (10 species) was registered on one specimen only. The parasite load was dispersed unevenly among the infected voles. Mean number of parasites of all species on a host individual varied from 124 to 295. The highest grade of parasites (237-297 parasite specimens) was found in the voles with 7-10 species of parasites. PMID- 12624961 TI - [Forest tick Ixodes ricinus (Ixodidae) in the foci of tick-borne borreliosis of the northwestern part of the Moscow region]. AB - Monitoring studies during 1996-2001 have shown a natural foci of tick-borne borreliosis with high abundance of Ixodes ricinus and high infection rate with an agent: 19.5-38.4 tick individuals per 1 km of route, infection rate 18.0 +/- 1.7 22.5 +/- 1.5%. A risk to be infected with the Lyme disease in the ricinus focus of the tick-borne borreliosis during the epizootic season did not vary much during the period of study, 3.5-8.2 in different year, mean 6.0 infected ticks per 1 km of route. PMID- 12624962 TI - [Morphological barrier in mechanisms of reproductive isolation acting in areas of sympatry of closely related species Ixodes persulcatus-I.pavlovskyi and I. persulcatus-I. ricinus (Ixodidae)]. AB - A morphometric analysis of different male and female organs used in the process of copulation, including an attachment of the spermatophore to the genital opening of female and placing the spermatophore into the vestibular part of vagina, was carried out for the pairs of ixodid tick species Ixodes persulcatus- I. pavlovskyi and I. persulcatus--I. ricinus in areas of their sympatry. In two disjunctive areas of sympatry of I. persulcatus--I. pavlovskyi (South-East Altai and Southern Primorye), the morphological barrier is displayed in the precopulation mechanism of reproductive isolation and does act only in the pair I. pavlovskyi female--I. persulcatus male. It is proved by the reliable prevalence of the hypostome diameter (in widest part) of I. persulcatus male over the genital opening length of I. pavlovkyi female. Because of this reason, the copulation between these partners cannot take place. On the other hand, there are no any morphological obstacles for a copulation within each species examined and in the combination I. persulcatus female--I. pavlovskyi male, and in any combination of the partners I. persulcatus--I. ricinus in sympatry areas (samples from two locations in the north-west of the Eastern-European Plane have been examined). Morphological peculiarities causing differences in host preference of tick species, that in turn realizes the isolation mechanisms between species, are discussed. Postcopulation non-genetic components of the reproductive isolation mechanism have been found in examined species. PMID- 12624963 TI - [Parthenogenetic generations of Sanguinicola armata (Trematoda: sanguinicolidae)]. AB - Daughter sporocysts of Sanguinicola armata are represented by several generations, changes of which goes synchronously with the changes of year seasons. Young individuals beginning the reproductions form exclusively cercariae. The old sporocysts begin to produce sporocysts only. These young sporocysts do not quite the organism of the old sporocyst. Therefore, series of sporocysts inside other sporocysts are often observed in hystological cross sections. Germinal masses of daughter sporocysts of S. armata have some specific characters, which are not observed in analogous organs in daughter sporocysts of other trematode species. PMID- 12624964 TI - [Parasitic systems of Microsporidia: descriptions and terminology questions]. AB - Three parasitic systems of Microsporidia are described: the system of monoxenic Vairimorpha mesnili with paraxenic hosts presented lepidopteran and hymenopteran species; the system of dixenic Amblyospora sp. with metaxenic hosts presented bloodsucking mosquitoes and crustaceans and the system of Metchnikovella sp. as parasite of other obligate parasite. The last case is characterized by very intimate interrelations between hyperparasite (microsporidian species), obligate parasite--host of Microsporidia (gregarine) and hyperhost--host of gregarine (polychaeta). This hyperparasite system is exclusive case of parasitic systems. Parasitic and hyperparasitic systems reflects a population level of host-parasite interactions. On biocenotic level many other organisms such as predators, vectors and reservators of invasion stages of Microsporidia affect parasitic systems giving a chance to one of the members of the system (to the host or to the parasite). These organisms form epiparasitic system. In all cases of the parasitic systems there are two-way communications between parasites and their hosts. In systems on biocenotic level--parasitic consortium--members of epiparasitic systems acts on parasitic systems, but members of parasitic systems don't affect epiparasitic systems. PMID- 12624966 TI - [Myxosporeans (Cnidospora: Myxosporea) of Antarctic fishes]. AB - Data on 13 species of fish myxosporeans from the Antarctic part of the Atlantic Ocean is present. Among them, 8 new species are described: Zschokkella australis sp. n., Davisia nototheniae sp. n., Ceratomyxa orthospora sp. n., C. ellipsoidea sp. n., C. antarctica sp. n., Leptotheca ampla sp. n., Pseudalataspora squamifrons sp. n., P. meridionalis sp. n.). Four species were found for the first time in the region and four species were found in new hosts. PMID- 12624967 TI - [Bradophilidae fam. n.--a new family of mesoparasitic copepod (Copepoda: Poecilostomatoida), collected from the polychaete Brada villosa from the Beloe more]. AB - The Bradophila pigmaea Levinsen, 1878 is redescribed on the basis of new material collected firstly after original description. The new family Bradophilidae belonging to order Poecilostomatoida is erected on the basis the genus Bradophila. The male of Bradophyla pigmaea is described in detail at firstly. PMID- 12624965 TI - [Peculiarities of metabolism of the microsporidia Nosema grylli during the intracellular development]. AB - A long adaptation of Microsporidia to intracellular development supposes the host derived ATP dependence of merogony and sporogony stages. To prove this assumption the activities of ten carbohydrate and energy metabolism enzymes were compared in the microsporidia Nosema grylli intracellular stages and mature spores. This species infects the fat body of crickets Gryllus bimaculatus. We have demonstrated lower activities of glycolytic enzymes, phosphoglucomutase and glucose-6-PhDH in the metabolically active meronts and sporonts than in the dormant mature spores. Low glycolysis level indicates that carbohydrate catabolism is not a principal mechanism of ATP supply in the N. grylli intracellular stages. Furthermore, we have not revealed a preferable expenditure of glycogen in comparison with triglycerides in infected cricket fat bodies. The N. grylli infection causes an equal reduction of glycogen and lipid content approximately in 2-3 times. Microsporidia have not mitochondria, Krebs cycle and electron-transport chain. Therefore they are not able to utilise fat reserves for ATP production. It seems to be proposed that microsporidia consume exogenous ATP which is produced by host cell metabolic system. The N. grylli infection provokes an increase of ATP content and ratio of ATP/ADP concentrations in cricket fat bodies approximately in 4 times. These data indicates a rise of host cell energy metabolism rate during the infection. PMID- 12624968 TI - [Safety management in surgery]. PMID- 12624969 TI - [New directions of patient safety policy in several foreign countries]. AB - The importance of safety in healthcare has long been recognized, but actual policies and actions to prevent medical errors and improve patient safety have not been developed comprehensively. Retrospective studies based on medical records provide evidence of the extent of adverse events in healthcare services. The new trends in policies for patient safety differ radically from the previous ones in that they emphasize the necessity of reforming the entire healthcare system to make patient safety the first priority. To achieve this it is necessary to establish an adequate information system to collect and analyze data in a timely fashion and distribute the results to stakeholders to encourage "Safety Culture" in healthcare industries. PMID- 12624970 TI - [Actions for improvement of medical safety by all parties involved in healthcare]. AB - How to prevent medical adverse events and augment the public's confidence in healthcare has become an urgent and one of the highest-priority medical policy issues in Japan. All parties involved in healthcare, such as medical institutions, companies that sell medical supplies and devices, educational institutions, and medical societies, are asked to work together cooperatively in their respective roles to ensure medical safety. Medical institutions, as the facilities that actually provide healthcare, are responsible for improving the safety and reliability of medical treatment. They must do this through appropriate supervision of their organizations and maintenance of their systems under the sound leadership of their management, and they must tackle safety measures by improving their organizations. Medical societies are expected not only to make scientific contributions but also to collect and evaluate information on the risks in their specialties and disseminate the information widely to their members. The national government must clarify the short- and long term goals for promoting medical safety. At the same time, it must maintain the appropriate environment and coordinate the involvement of concerned parties in the achievement of those goals. PMID- 12624971 TI - [Reporting of patient deaths and grave injury incidental to clinical practice]. AB - The Japan Surgical Society has developed guidelines for the reporting of patient deaths and grave injury incidentally occurring in their clinical courses, following its declaration in March 2001 in reference to practitioners' duty to notify the police under Article 21 of the Medical Act. These guidelines can be summarized into three points. First, they are ethical guidelines for practitioners and are not concerned with the legal duty required under the Medical Act. Second, they require reporting of patient deaths even if the cause is still not confirmed as medical malpractice, and reporting of grave injury due to apparent medical malpractice, not mention deaths due to apparent medical malpractice. Third, it is emphasized that the guidelines be applicable for the time being until some generalized system is established, which is specialized to accept reports about patient deaths and grave injury incidentally occurring in their clinical course from every hospital nationwide and to collect data concerning small incidents and risks in medical practice. The Japan Surgical Society strongly requests that such a general system start to function as soon as possible. In advance, the current guidelines should be amended repeatedly in reference to problems occurring in actual cases. PMID- 12624972 TI - [Patient safety management in surgery]. AB - This article deals with practical guidelines for informed consent to surgery and strategies to avoid adverse events during the operative and postoperative periods. It also describes what to do in the event of accidents and what should or should not be done to avoid medical litigation. PMID- 12624973 TI - [Safety improvement through implementation of clinical pathways]. AB - In Japan the clinical path (or critical path) is attracting attention as a tool for the improvement of healthcare in terms of optimization of informed consent, development of a team approach in healthcare, transformation of the healthcare system into a patient-centered pattern, economizing on medical resources, etc. As clinical pathways can contribute to the reduction of medical errors and the early detection of abnormalities, they are also very effective in promoting patient safety. Important aspects of risk management include gathering and analyzing past incident reports and establishing countermeasures; they imply that each individual health professional, including each physician and nurse, improve their medical knowledge with regard to risk management and that health professionals improve their communication skills and share as much information as possible. In addition, it is imperative for risk management to standardize medical care, streamline the components of healthcare, and avoid unnecessary tests and treatment measures. Clinical pathways are remarkably useful for all of these goals. Another important consideration is the fact that patients and their relatives can promptly identify irregularities arising from deviations from clinical path stipulations. Clinical pathways are highly useful with regard to risk management. PMID- 12624974 TI - [Safety management in operating rooms]. AB - In operating rooms, patients with a wide range of conditions are transferred from the wards to undergo surgery. Furthermore, because surgeons with different specialties perform various operations using instruments unique to their specialty, it is possible that operating room nurses, surgeons, and ward nurses do not always communicate clearly. Patients are anesthetized and unconscious, and as a general rule, family members are not allowed entry. As a result, operating rooms are unique environments where neither the patients receiving medical services nor their families can express opinions and concerns. Therefore an especially strict crisis management approach is needed in operating rooms. At present, individual hospital implement their own crisis management systems to prevent medical accidents. However, national debate is needed to establish medical accident prevention guidelines before each hospital compiles a manual that suits its needs. PMID- 12624975 TI - [Nosocomial infections and risk management]. AB - Nosocominal infections have been increasing due to the complexity of treatment modalities, more numerous admissions of elderly persons from nursing homes, and readmission of the patients with implanted devices. It is mandatory to have risk management systems in place in Japan to improve the quality of medical practice and also to avoid unexpected morbidity and mortality. This paper discusses the importance of surveillance for infection, control of infectious outbreaks, guidelines for antimicrobial usage, and protection of healthcare workers. PMID- 12624976 TI - [Safety management activities of Yokohama City University Hospital--safety management is the platform of quality improvement]. AB - Four years have passed since mistaking patients for operations occurred at Yokohama city university hospital (YCU hosp) on January 11, 1999. The accident was not just a simple mistake such as the patients happened to have the same name. Its root seemed to be a system error that deeply related to health care delivery at YCU hosp. Based on the trying experience, YCU hosp started to construct a new system for health care safety management. From 2001 the system development entered the second phase. The activities of this phase included promoting interdisciplinary projects on a variety of topics, implementing a systematic educational program, and activating risk managers. As a whole, autonomous activities by health care professionals regarding patient safety started to take place in the hospital as a new approach for safety management. The next step will be to expand these activities into a movement for hospital wide quality improvement and establishing clinical governance. PMID- 12624977 TI - [Diagnosis related groups (4). Future direction of DPC research and hospital services based on DPC]. PMID- 12624978 TI - Drug resistant-Mycobacterium leprae--results of mouse footpad studies from a laboratory in south India. AB - Out of 265 biopsies of leprosy patients received at the Experimental Pathology Laboratory of Schieffelin Leprosy Research and Training Centre from 1987 to 1997 for evaluating resistant strains of M. leprae, using the mouse footpad technique, 49 showed resistant strains of M leprae to varying concentrations of dapsone, rifampicin and clofazimine. 23 (47%) of these were from a control area. With 369 skin-smear positive multibacillary (MB) patients as the risk group (denominator), 23 (6.23%) were resistant to one or more drugs. 18 (4.88%) had dapsone resistance, 5 (1.36%) were resistant to rifampicin and 9 (2.44%) had resistance to low concentrations of clofazimine (0.0001%). Out of the 23 biopsies with drug resistance from the control area, primary dapsone resistance was seen in 7 (30%) biopsies and secondary dapsone resistance in 11 (48%). Primary rifampicin resistance was seen in 4 (17.4%) patients, secondary rifampicin resistance in 1 (4.35%) and primary clofazimine resistance in 7 (30%). 3 (13%) of the strains showed secondary clofazimine resistance. One biopsy had resistant strains to all the three drugs. In a control area where properly supervised effective multidrug therapy (MDT) was regularly administered over the years, the emergence of drug resistance is negligible. It may not be the case if the content, duration and regularity of the drug regimen were not satisfactory. Aware of the possible shortcomings in mass administration of MDT, it is emphasized that mouse footpad studies on drug resistance should be made available at least in endemic areas where the incidence of the disease has not changed despite good MDT coverage in order to monitor the emergence of drug resistance. Research into molecular biological identification of drug resistant-M.leprae should be intensified. These steps would help to institute timely measures to check the spread of any drug resistant organisms in the community. PMID- 12624979 TI - Multidrug therapy in leprosy can prevent relapse--a retrospective study. AB - A retrospective study was done at the Leprosy Control Unit (LCU) in Durgapur of Burdwan district, West Bengal, to determine the relapse rate following multidrug therapy (MDT). A total of 1581 patients (1276 PB and 305 MB) completed MDT regimens during a period of 5 years as per WHO recommendations and National Leprosy Eradication Programme (NLEP) guidelines. The treated patients were kept under surveillance as per NLEP guidelines and searched for relapses. The results of MDT were compared with those of pre-MDT (monotherapy) era at the same centre (total: 405 patients; PB-373, MB-32) and also with those of the Leprosy Clinic in Gopalpur (only dapsone was given to a total of 189 patients, PB-167, MB-22). Following monotherapy, the relapse rate was 10.06% at the Gopalpur Leprosy Clinic and 12.4% at the Durgapur LCU during the 2 years (PB) and 5 years (MB) of surveillance, whereas following MDT no relapse case was encountered both in PB and MB cases during the surveillance periods recommended by WHO. The results of this study are comparable with those of other studies. Though a few studies showed relapses during long-term surveillance beyond the periods recommended by WHO, it is once again established that MDT can prevent relapse in leprosy. PMID- 12624980 TI - Grip and pinch strength in relation to function in denervated hands. AB - A study was conducted in the Schieffelin Leprosy Research and Training Centre, Karigiri, Tamil Nadu, from March to September 2000 to find out the relationship between grip and pinch strengths and basic activities of daily living (BADL) in leprosy patients. Sixty-two patients (mean age: 36.1 years) suffering from impairment of motor nerve function in one or both hands were included in the study. Grip strength and pinch strength, namely pulp-to-pulp pinch strength, lateral pinch strength, tripod pinch strength, were measured using a hydraulic hand dynamometer and a pinch gauge. BADL was assessed using Karigiri activities of daily living rating scale. Frequency table, mean, median, range, standard deviation, correlation and multiple regressions were used in analysis. The results showed that there was a highly significant correlation (p < 0.001) between the grip and pinch strength and BADL. Multiple regression analysis showed only tripod pinch strength had a significant, independent influence on the BADL. Other two pinch strengths and grip strength were not significant. These results suggest that there is a significant relationship between the grip and pinch strengths and BADL. The tripod pinch strength is the most important hand strength contributing to the problems in BADL. In a measure of association between impairment and disability, the results revealed that out of 62 patients with physical impairment of the hand, 59 (95.2%) had experienced disabilities. This indicated that, despite impairment, 4.8% do not have any disabilities. PMID- 12624982 TI - An interventional strategy to strengthen integration of NLEP into primary health care. AB - The Government of Orissa implemented the Revised Operational Strategy in September 1999 to integrate the NLEP functions into primary health care activities. An interventional strategy, in the form of consensus on job responsibilities and capacity-building through training of PHC staff, was developed and adopted in a rural block under the Department of Community Medicine to strengthen the integration process. The impact was studied six months after the intervention by comparing it with the leprosy situation in the pre intervention period. Data were collected by verification of registers at the block PHC and sub-centre levels. Analysis was done using different leprosy indices, such as new case-detection rate (NCDR), child rate, deformity rate, profile of leprosy cases and patient compliance, etc. This integrated approach was found to be more community-oriented and effective in early case-detection in children and women. It also helped in providing continuous MDT services because of the involvement of primary health care functionaries in the post-intervention period. PMID- 12624981 TI - Does pentoxifylline find a place in the armementarium of leprologists in type II reaction? AB - Alternative therapeutic interventions in Type II lepra reaction are being considered following serious problems associated with the use of steroids and thalidomide. Pentoxifylline (PTX) has been used in Type II reaction with varying degrees of success. The results of a study on the use of this drug in a dose of 1200 mg per day for a period of 2 months in patients with ENL reaction are discussed. Five patients, one of whom was HIV positive--all with severe Type II reaction, were regularly evaluated for regression of inflammatory symptoms and clinical involution of ENL lesions while on PTX therapy and thereafter. It was found that PTX led to a total elimination of systemic symptoms within a week. ENL lesions regressed in two weeks. However, in one patient, lesions recurred after one month of therapy. It appears that PTX is well tolerated and could be used as an additional drug in the armamentarium of leprologists in the management of Type II reaction, especially in HIV co-infection, where long-term steroids are contraindicated. However, further studies to compare the effects of PTX with currently, widely used drugs for the treatment of ENL reaction are necessary. PMID- 12624983 TI - Follicular mucinosis in suspected leprosy. AB - Four cases of suspected leprosy showed, on biopsy, follicular mucinosis without any granulomatous inflammation. All the patients were adolescents (12-17 years) with a single lesion on the face. Three patients showed complete clearing after anti-leprosy treatment, and the fourth patient is currently taking anti-leprosy treatment and showed good clinical response. PMID- 12624984 TI - [Influence of thinning on regeneration in a coastal pinus thunbergii forest]. AB - A coastal forest planted nearby the sea can provide many shelter benefits for the coastal regions. It is ideally if the continuity of the shelter benefits could be preserved through reasonable management. Thinning and regeneration as the most important management techniques for plantations can help the continuity of the shelter benefits of the coastal forest. However, because of the peculiarities of coastal forest, i.e., the coastal plantation nearby the sea is vulnerable to disturbances (thinning as one kind of disturbance), the study on thinning and regeneration within a coastal forest is poorly understood. The purpose of this paper is to give a primary understanding in natural regeneration for the coastal Pinus thunbergii forest with different thinning rates after four growing seasons since thinning. The experiment was carried out at the middle of the shoreline along the Japan Sea, and the investigated sites consisted of four thinning treatments (control, 0% thinned, 20% thinned, 30% thinned and 50% thinned) in a coastal P. thunbergii forest. After thinning, the regenerated seedlings, soil water content, light condition (canopy openness or canopy density), wind regime, and litter depth and quantity were investigated for four growing seasons. The relationships between the regenerated seedlings and light condition, litter, wind profile and soil water content were examined. The results showed that thinning could improve the light condition on the forest floor, increase the exchange of airflow (wind speed) in the coastal forest stand, and ameliorate the water content of the forest soil. These factors accelerated the decomposition of litters, and provided necessary conditions for natural regeneration. The results of regeneration observation indicated that the most intensively thinned treatment (50% thinned with density of about 1500 stems.hm-2) could provide a better condition for regeneration during the four growing seasons. The density and growth of seedling (greater than 1 year) increased significantly with increasing thinning intensities, and the establishment of seedlings was obviously succeeded in the most intensively thinned treatment, but failed in less thinned treatments and understory. The thinned intensity of 50% did not induce wind damage to the coastal forest in the four years after thinning, and did not cause the loss of shelter functions of the coastal forest such as sand blocking, wind breaking and salt preventing etc.. On the contrary, it could provide the suitable conditions for natural regeneration of the pine coastal forest, or for the immigration of other species. Therefore, thinning as the silviculturally created openness is very important for the establishment of seedlings in the coastal forests, which provides a mechanism for the coastal forest from even aged stands dominated by P. thunbergii to stands containing multiple size classes and canopy layers. PMID- 12624985 TI - [Population dynamics of beauveria bassiana in masson pine plantation ecosystem]. AB - Annual dynamics of Beauveria bassiana in 12 experimental plots of masson pine plantation inoculated at different doses and frequencies was investigated. The results showed that inoculum density of B. bassiana in the plantation was correlative to released inoculum in initial stage after B. bassiana released. In this period, the amount of inoculum densities at different niches of the plantation was in order of litter layer > crown layer > soil layer. Four months later, inoculum densities in X-plots (2 releases a year), Y-plots (1 release a year) and Z-plots (1 release every two years inoculated one year ago) fluctuated stably within certain ranges. Therefore, the optimal frequency and dosage of inoculation releasing B. bassiana was determined as twice a year, and 37.5 g.hm-2 (10(11) conidia.g-1) each time. PMID- 12624986 TI - [Distribution pattern of rare plants along riparian zone and its implication for conservation in Shennongjia area]. AB - Due to the importance of riparian zone in maintaining and protecting regional biodiversity, more and more ecologists paid their attentions to riparian zone, and had been aware of the important effects of riparian zone in basic study and practical management. In this study, forty sampling belts (10 m x 100 m) parallel to the bank of Xiangxi River at different elevations in Shennongjia area were selected to investigate the riparian vegetation and rare plants. Fourteen species of rare plants were found in riparian zone, accounting for 42.4% of total rare plant species in Shennongjia area. The main distribution range of the fourteen rare plant species was the mixed evergreen and deciduous broadleaved forest at elevation of 1200-1800 m, where species diversity of plant community was the maximum at the moderate elevation. Fourteen rare plant species could be divided into three groups against the elevation, namely low elevation species group, moderate elevation species group, and high elevation group. In the paper, the authors discussed the reasons forming the distribution pattern of rare plant species, and pointed out the important function of riparian zone on rare plant species protection. PMID- 12624987 TI - [Mating system and inbreeding retrogression of casuarina equisetifolia plantation, an introduced species in Xiamen]. AB - Casuarina equisetifolia, an introduced species, is a good shelter species planted along coastlines due to its enduring sandy and saline habitats. Decline was, however, observed in C. equisetifolia plantations in many areas since 1980's. Mating system and inbreeding retrogression of a plantation of C. equisetifolia in Xiamen and their roles in the decline of C. equisetifolia forests were thus researched using allozyme technique. Multilocus outcrossing rate was 0.622, lower than that expected from related species, indicating a mixed-mating system, and more inbreeding caused through this introduction. Calculations from electrophoresis data showed that very a high inbreeding retrogression occurred in the C. equisetifolia plantation, indicating that inbreeding an its subsequent retrogression played an important role in the decline of C. equisetifolia plantation. Alleviating the decline through genotype species-introduction was also proposed. PMID- 12624988 TI - [Distribution of anthraquinones in leaves of two Aloe species and their defence strategy]. AB - Aloe plants are the succulents mainly distributed in arid or semi-arid desert in South Africa. TLC analysis indicated that Aole arborescens and Aloe hereroensis contained the high concentration of phenolic derivative metabolites, anthraquinones such as barbaloin, homonataloin, aloeresin and aloenin. In younger leaf, L3 of A. hereroensis, the average content of 4 anthraquinones reached 44.9% of the dry weight of exudates. The similar distribution of the anthraquinones in the two species were found, which showed that the youngest leaves had the highest content, the top part of each leaf had the highest content, and the basal part had the lowest content. Along leaf margin, the content was higher than that of central parts. However, some different distributions were also revealed and compared between the two species. It was inferred that the special distribution of anthraquinones in Aloe was a chemical defense strategy for protection themselves. PMID- 12624989 TI - [A comparative study on drought-resistance of four plant species in kerqin sandy land]. AB - The water physio-ecological index and photosynthetic rate of four plant species, Caragana microphylla, Amorpha fruticosa, Artemish halodendron, and Lespedeza bicolor, in kerqin sandy land were determined. The results showed that C. microphylla has the lowest leaf water potential and the highest water maintainance, bound water/free water ratio, and water utilization efficiency. Its drought-resistance is the strongest. Amorpha fruticosa was contrary to Caragana microphylla, and its drought-resistance was the weakest. Artemish halodendrona had a high leaf water potential, but its water-maintainance, bound water/free water ratio, and water utilization efficiency were very high. Its drought resistance was stronger. Lespedeza bicolar had a lower leaf water potential and water-maintainance, and a usual bound water/free water ratio, and water utilization efficiency. Its drought-resistance was stronger than Amorpha fruticosa's, but weaker than Artemish halodendron's. PMID- 12624990 TI - [Ecological benefits of artificial seabuckthorn stands in semi-arid hilly region of Loess plateauion soil- and water conservation and soil moisture]. AB - There is a remarkable function on decreasing runoff and sediment in seabuckthorn and its mixed stands, but the effects on soil- and water conservation are different due to different structure and patterns of seabuckthorn and its mixed stands. The intensity of soil water use by seabuckthorn forest was different along with the month in growing season. In growing season, soil moisture in 0-500 cm layer was 5.1%, the lowest in the end of May and 8.8%, the highest in the end of October. This intensity was also different with forest age. 8 ages seabuckthorn forest consumed 231.2 mm of soil stored water in 0-500 cm layer annually, and soil moisture was 5.6%. Therefore, it should be cut for increasing soil moisture. In end of the third year after cutting, soil moisture in 0-160 cm layer could recover, and the value would be 10.3%-14.6%. Seabuckthorn forest could reduce the effect of slope location on soil moisture. Seabuckthorn and its mixed stands have similar intensity of soil water use, and there was a soil dry layer phenomenon in their stands. PMID- 12624991 TI - [Simulating photosynthesis and evapotranspiration of winter wheat with a SVAT model]. AB - Two sub-models were added to the developed soil-vegetation-atmosphere transfer model. Of the two sub-models, one was the canopy photosynthesis/stomatal coupled sub-model distinguishing the sun light and shaded leaves, and the second was the simplified growth sub-model considering growth respiration and maintenance respiration. The comparison of measured and simulated daily evapotranspiration showed that the RMSD (root mean standard deviation) of model prediction to observation was 0.65 mm, and MAPD (mean absolute percent difference) was 14%. The daily course of the simulated and measured photosynthesis rates at the top of canopy were in good consistency. With the high-resolution meteorological data, the canopy net photosynthesis and evapotranspiration were simulated over the whole growing season. The sensitivities of maximum leaf area index, environmental CO2 concentration and leaf nitrogen content to evapotranspiration and water use efficiency were also analyzed. PMID- 12624992 TI - [Effect of soil water on the growth and physiological characteristics of grafted and non-grafted cucumber in greenhouse]. AB - Studies on the effect of soil water on the growth and physiological characteristics of cucumber in greenhouse showed that water deficit or excessiveness could retard the growth and development of cucumber. The transpire rate, Gs, water potential and root activity decreased quickly due to water deficit. PMID- 12624993 TI - [Developmental genetic analysis of panicle dry weight of indica hybrid rice oryza sativa under different environmental conditions]. AB - Four indica sterile lines (A) and corresponding maintainer lines (B) as well as five indica restorer lines, which varied in panicle dry weight, were used to create genetic populations of two generations according to incomplete diallel cross design (4 x 5). Analysis on panicle dry weight at different developmental stages based on tow-years experimental data was conducted by using additive dominant developmental genetic models with genotype effects and genotype x environment interaction effects. The results showed that panicle dry weight was controlled by both genetic main effects and genotype x environment interaction effects in the whole developmental process. During the early-middle developmental periods (15 days after flowering), panicle dry weight was mainly governed by dominant effects, and dominant genes expressed with large amount. The meanwhile, positive effects produced by environmental factors appeared to be significant. Proper cultivation measure to create good external environment conditions for rice panicle was helpful to promote the expression of heterosis potential completely. Within the middle-late developmental periods (from the fifteenth day to the thirtieth days after flowering), additive effects played major role, and its genes expresed in the most active state. Genetic selection for panicle conducted in this period was expected to gain better genetic advancement. PMID- 12624994 TI - [A long-term field trial on fertilization and on use of recycled nutrients in farming systems III. Recycling rate of P and N through a composting cycle and recoveries of P and N in organic manure]. AB - The results from a nine years experiment indicated that the average loss rate of N and P contained in harvest products through a feeding-composting cycle was about 30%. Under high yield conditions, the amount of recycled N and P from 60% of harvested products and through a composting cycle in a farming system were about 55-70 kg.hm-2 and 9-11 kg.hm-2, respectively, equivalent to 37%-47% of N and 36%-44% of P from fertilizers applied every year to the system. PMID- 12624995 TI - [Changes of crop yield and soil fertility under long-term application of fertilizer and recycled nutrients in manure on a black soil III. Soil nutrient budget]. AB - The nutrient budget of fertilization models under different treatments was calculated using data from a field experiment over the period of 1985-1999. The results indicated that application of nitrogen fertilizer accelerated a large deficit of soil phosphorus, and the use of nitrogen and phosphorus accelerated the deficit of potassium. The experimental data demonstrated the appearance of a large area of soil deficit of phosphorus from 1970s and of potassium from 1980s in China. Nutrient recycled in farming system improved soil nutrient budget, but could not meet the nutrient requirements from high-yield crops. The use of recycled nutrients with an appropriate use of fertilizers according to the soil fertility could produce higher crop yields, balance soil nutrient budget, and not cause surplus nutrients to emit into environment. PMID- 12624996 TI - [A long-term field trial on fertilization and on use of recycled nutrients in farming systems IV. Soil fertility changes]. AB - The fertility changes of soil under different fertilization treatments of a twelve years field trial indicated that the application of compost recycled in a farming system with appropriate amount of fertilizer applied to balance the soil nutrient budget, could not only produce higher crop yields and less surplus of nutrients into environment, but also improved soil fertility with significant increase of soil organic carbon and nitrogen in cultivated layer. However, maintaine higher soil available phosphorus, it was not sufficient just to balance the budget of soil phosphorus, and more phosphorus fertilizer application was needed. PMID- 12624997 TI - [Effects of neem seed extracts on nitrogen use efficiency in two different soils]. AB - Incubation test and pot experiments were conducted with haplic luvisols and hydragric anthrosols to study the effects of neem seed extracts (N I, N II) on nitrification and immobilization of ammonium sulfate. N I could significantly inhibit the nitrification of N applied to the two soils. N II was effective in promoting the immobilization of NH4+(-)N. Pot experiments showed that N II could increase the use efficiency of chemical nitrogen significantly in fimic anthrosols. PMID- 12624998 TI - [Effects of phosphorus application methods on phosphorus loss on sloping land under simulated rainfall]. AB - A simulated rainfall experiment was conducted to study the effects of three phosphorus application methods on phosphorus loss and its forms in loss process. The results showed that the bio-available P (BAP) loss in runoff was positively correlated to the mixed intensity of P applied into soil. Under mixed application (MA), the dissolved phosphorus (DP) concentration, BAP concentration, DP/BAP and BAP/TP ratio in runoff were all the highest, and those under point application (PA) were the second. Under line application (LA), DP and BAP concentrations were low, DP/BAP and BAP/TP ratio were also relatively low, and the difference compared with control was insignificant. According to the reduction of P loss, the order of the three P application methods was LA > PA > MA. PMID- 12624999 TI - [Effect of iron oxides addition on organic acids content in paddy soil]. AB - The addition of ferrihydrite into paddy slurries significantly reduced the concentration of acetate. In fresh paddy soil, due to the competitive consumption for acetate by addition of ferrihydrite, the concentration of acetate was reduced to 10-20 mumol.L-1 after 5d anaerobic incubation. However, in control slurries, the acetate concentration was still as high as more than 1200 mumol.L-1. In soil produced low acetate, the acetate was completely consumed by addition of ferrihydrite, resulting in a full inhibition of methane formation. Also, the application of lepidocrocite reduced methane genesis, but the reductive effect was less than ferrihydrite. The acetate was largely accumulated during a short term anaerobic incubation from 1 to 10 days by hematite addition, but at the same time, the methane genesis was not extensively increased. The propionate concentration was also reduced under anaerobic incubation by addition of different iron oxides such as ferrihydrite, lepidocrocite and aluminum substituted goethite, and the inhibitory effect was in order of ferrihydrite > lepidocrocite > Al-substituted goethite. The types and contents of organic acids in fresh soil were greatly different from those being incubated for 11 weeks under an anaerobic condition. PMID- 12625000 TI - [Effects of probiotics additives EM on growth metabolism and immune function of blue fox in growth phase]. AB - Different doses of probiotics EM were added to the diets of blue fox in its growth phase to study the effects of EM additives on its growth, digestion metabolism and immune function. The results showed that with EM additives applied, the ingestion and average daily gains were increased remarkably (P < 0.05). The apparent digest rate of crude fibre, crude protein, mineral elements Ca, P and NPU could be increased (P < 0.05) and correlated with its dose, and the digest rates of the later four targets in group III were increased by 8.21%, 16.27%, 9.82%, and 12.13%, respectively. EM additives could not increase the digest rate of ether extral and N-free extract. Disease of diarrhoea and blood in stool could be prevented and cured, and immune function and the survival rate of fox in growth phase could be increased remarkably. EM, which a kind of efficient, safe and environmental protection additives, can be extended and applied in farming of fur animals. PMID- 12625001 TI - [Role of contagious infection of Pandora delphacis in suppression of Myzus persicae colonies]. AB - The colonies of green peach aphid, Myzus persicae, on detached cabbage leaves (8 replicates) were initiated with each including 3 apterae exposed to 'spore shower' of the entomophthoraceous fungus, Pandora delphacis, to evaluate its potential for aphid control. The colonies were then allowed to freely propagate and infect from one to another for mycosis development at different regimes of temperature (10-30 degrees C) and relative humidity (74-100% RH). During a period of 30-day observation, aphid mycosis developed much more rapidly at the regimes of higher temperature (20-30 degrees C) and humidity (> or = 95% RH), at which, nymphs were effectively infected by contacting the conidia discharged from apterous cadavers. The efficacy of P. delphacis for control of M. persicae was easily visible at all regimes considered, despite variation in mycosis-caused mortality. Compared to the increase of M. persicae colony not contaminated with the fungal agent, the efficacy of control at all humidity regimes of 30 degrees C was the best, and it could be > 60% on day 4, and 100% on day 16. Secondary to the best, the increase of colony size at 20 and 25 degrees C was controlled by > 30% on day 8, and > 80% on day 20 at all the humidity regimes with occasional exceptions. The efficacy of control at 10 and 15 degrees C was usually inferior to those at higher temperatures, but to less degree associated with relative humidity. These results indicate that P. delphacis was of high potential for aphid control, deserving further study for practical utilization. PMID- 12625002 TI - [Dynamics of predatory natural enemy community in cotton fields]. AB - The dynamics of predatory natural enemy community in cotton field along the Yangtze River in Anhu province was studied. The results showed that there were totally 19 species of predatory natural enemies, and the dominant species were Propylaea japonica, Erigonidium graminicola, Theridium octomaculatum, Misumenops tricuspidatus, Orius minutes and Geocoris ochropterus. The predatory natural enemies in the cotton fields were rather abundant in the whole growing period of cotton, except the rainy season. Among the dominant species, P. japonica, M. tricuspidatus, O. minutes and G. ochropterus were distributed mainly in the upper and middle parts of cotton plants, while E. graminicola and T. octomaculatum mainly in the middle and lower parts of cotton plants. The diversities of the community were relatively high in the early developmental period of cotton, and reached the top level in the bud and flowering period of cotton in the first 10 days of July, then declined successively hereafter. The principal factor affecting the diversity was species richness in the early developmental period of cotton (from shoot period to bud and flowering period), and species evenness in the late developmental period of cotton (from flowering and boll period to boll opening period). The characteristics of the community were relatively analogous before the second 10 days of August (from shoot period to flowering and boll period of cotton), but after this period, the characteristics changed obviously with the features of the dominant species getting more and more conspicuous. PMID- 12625003 TI - [Starvation metabolism in parent Chinese mitten-handed crab (Eriocheir sinensis)]. AB - The experiment was conducted to study the influence of starvation on the metabolism of parent Chinese mitten-handed crab (Eriocheir sinensis) with weights of 54.59 (+/- 2.37) g under 20 (+/- 0.5) degree C during Oct. to Dec., 1998. The results showed oxygen consumption, CO2 production and ammonia-N excretion of the parent crab being starved for 30 days were 50.0%, 63.4%, and 59.1% of those of the fed crab, respectively. The course of oxygen consumption reduction could be divided into four phases: 1) from the 1st day to the 6th day, the metabolic rate decreased remarkably; 2) from the 7th day to the 15th day, the metabolic rate kept relatively stable; 3) from the 16th day to the 25th day, the metabolic rate decreased markedly again; and 4) from 26th day to 30th day (the end time of the experiment), the metabolic rate decreased most dramatically. There were only three phases during the course of CO2 production and ammonia-N excretion reduction. Meanwhile, the standard metabolism of the starved crab reduced from 4.45 to 2.36 J.g-1.h-1, and lipid was used as the first energy source in the course of starvation. PMID- 12625004 TI - [On nest-site selection of Otic tarda]. AB - During April to July in 2000 and 2001, the nest-site selection of Otis tarda was studied by direct observation and sampling method in the southeast grassland of Keyouqianqi in Inner Mongolia. The results showed that Otis tarda was one kind of big-sized bird built nests on the ground in sparse cluster, and the smallest distance between nests was 9 m. Otis tarda had a significant tendency in nest site selection. Firstly, this tendency was represented under the scale of landscape. Most nests of Otis tarda were built on the mid-hill of grassland, the elevation range was from 190 m to 230 m, most nests built on the slopes faced to sun, and the biggest gradient was not more than 8 degree. Moreover, the nests were near to roads. Secondly, Otis tarda selected its nest-site according to vegetation structure. It tended to select nest-site with thick hay, high vegetation density and grass height from 15 cm to 35 cm, but would not select the place with over high vegetation density no nest. PMID- 12625005 TI - [Natural increase of parasitoids population of diamondback moth Plutella xylostella under ecological control condition]. AB - The parasitoids of diamondback moth Plutella xylostella was unfolded year after year since a series of ecological measurement was taken in Shenzhen Longgang Demonstrate Ecological Farm. The parasitical eggs of diamondback moth by Trichogrammatoidea bactraz and Trichogramma ostrinae were firstly found in the field in September 1999. Another two parasitoids, Trichogramma confusum and Trichogramma bilingensis also had a certain parasitism in nature. Now, the four Trichogramma species had already established their natural population, and their average parasitism was 15% around the year, and reached 25-30% at the harvesting period in Autumn, among them, Trichogramma confusum was predominant in quantity, which showed that ecological control played an important role in augmenting and protecting the population of Trichogramma in nature. PMID- 12625006 TI - [Microbiological eco-regulation in Vc two-step fermentation]. AB - The growth and interaction of Gluconobacter axydans and Bacillus megaterium in Vc two step fermentation have been studied. The results showed that there is a kind of coordinate symbiosis between G. axydans and B. megaterium during the fermentation, which could promoted 2-Keto-L-gulonic acid (2KGA) synthesis. The efficiency on the synbiosis would vary as the conditions of fermentation were different and were improved by the environmental factors. PMID- 12625007 TI - [Bio-remediation techniques of crude oil contaminated soils]. AB - The bioremediation of soils contaminated by different types of petroleum were carried out with composting process in a prepared bed. By the measures of nutrient- and microbiological agent addition, and moisture- and pH control, an ideal environment for microbes were obtained. When total petroleum hydrocarbons, which consist of thin oil, high condensation oil, special viscous oil, and viscous oil, were in the range of 25.8-77.2 g.kg-1 dry soil, the petroleum removal rate could reach 38.37-56.74% by 2 months operation. The contents of aromatic hydrocarbon, asphaltum and resin were important factors controlling the degradation of petroleum. 6 fungi, 6 bacteria and 1 actinomyces were found to be the dominant strains for petroleum degradation. The results could provide theoretical bases for remediation of soil contaminated by petroleum. PMID- 12625008 TI - [On nitrogen transformations and maturity during composting of pig manure]. AB - The contents of T-N, the ratios of soluble NO3-N/soluble T-N and soluble organic N/soluble T-N increased after co-composting for 63 days of pig manure and sawdust with or without additional leaves, while the ratios of soluble NH4-N/soluble T-N decreased. In this study, addition of leaves was beneficial for maturation of pig manure composting, and it could shorten the composting time from 42 d to 35 d. The ratios of soluble NH4-N/soluble T-N, soluble NO3-N/soluble T-N and soluble organic N/soluble T-N correlated with seed germination index (GI) significantly, and it could be used for maturity evaluation. PMID- 12625009 TI - [Effects of submarine topography and water depth on distribution of pelagic fish community in minnan-taiwan bank fishing ground]. AB - According to the fishing record of the light-seine information vessel in Minnan Taiwan bank ground during 1989 to 1999, the effects of submarine topography and water depth on distribution of pelagic fish community in Minnan-Taiwan bank fishing ground was studied. The results showed that the pelagic fish distributed concentratively, while the submarine topography and water depth varied widely, but in different fishing regions, the distribution of pelagic fishes was uneven. The distribution of fishing yield increased from north to south, and closed up from sides of the bank to south or north in the regions. Pelagic fish distributed mainly in mixed water in the southern Taiwan Strait, and in warm water in the Taiwan Strait. The central fishing grounds were at high salt regions. Close gathering regions of pelagic fish or central fishing ground would be varied with the seasonal variation of mixed water in the southern Taiwan Strait and warm water in the Taiwan Strait. Central fishing ground was not only related to submarine topography and water depth, but also related to wind direction, wind power and various water systems. In the fishing ground, the gathering depth of pelagic fish was 30-60 m in spring and summer, and 40-80 m in autumn and winter. PMID- 12625010 TI - [Preparation of PSAS and its application in HAB prevention]. AB - In order to obtain optimum PSAS (Polysilicate-aluminium sulfate) preparation conditions for removing HAB organisms, the 3-factor and 3-level orthogonal test was devised. Concentration of SiO2, Al3+/SiO2 molar ratio and pH were chosen as the three chief factors in PSAS preparation. The experimental results showed that the optimum preparation condition for various HAB organisms was different, and according to practical experimental operation, the appropriate preparation condition was considered as: concentration of SiO2 = 2%, molar ratio of Al3+/SiO2 = 1, and pH = 4. In addition, the coagulation efficiencies of PSAS on various HAB organisms were studied and compared with classical inorganic flocculant AS (Aluminium sulfate). The results indicated that the removal rates of PSAS were much higher than those of AS, and the dosages of PSAS were 30-40% lower than those of AS when they achieved the same removal rates. On the other hand, the anti-coagulation ability of HAB organisms varied with algal species, which was related to different physiological and ecological features of various algal cells. PMID- 12625011 TI - [Ecological restoration technologies for mined lands: a review]. AB - Mining activities usually cause catastrophic and extensive environmental changes, and eventually cause major damages to the whole ecosystem. The natural restoration for mine lands and tailings is a very slow process, and even can hardly reach their original states. Therefore, how to develop rapid and efficient approaches to accelerate restoration of mined lands has been highlighted by restorationists and environmental engineers during the past two decades. Almost all studies in this field indicate that the major problems come from soils: such as high metal concentrations, extremely strong acidity resulting from oxidation of pyrite, and poor fertility. Replacement of topsoil is therefore regarded as the most efficient method to alleviate adverse conditions of substrates; if this method is not available, other alternatives with lime, fertilizers, organic manures, garbage, mining wastes, and others will be applicable. In the aspect of using plants, species with strong resistance and rapid growth, like grasses and herbaceous legume, are always the first choice. If utilizing plants for the purpose of phytoremediation, species that are capable of accumulating exceptionally high concentrations of phytotoxic metals and of course, have a huge biomass, are preferably considered. No matter what type of ecosystem a mined land is restored or reclaimed to, an evaluation on whether it is a successful restoration or reclamation should be given. However, more practical, simple, and universal evaluation methods as well as more cost-effective, and operation-easy restoration techniques are still waiting to be developed. A set of artificial restoration methods that can be widely applied was summarized, and a discussion on the advantage and disadvantage of several evaluation systems was conducted in this review. PMID- 12625012 TI - [Research advances in soil persistence characteristics of toxins released by transgenic bt crops and their relationships with soil properties]. AB - With the popularization and application of transgenic Bt crops, and increasing attention has been paid to their potential impact on ecological environment and other aspects. And, as the development of detecting techniques for Bt toxin protein and soil were made by oversea researchers. This paper reviewed the current situation of the studies on the insecticidal characteristics and detecting methods of toxin proteins released by transgenic Bt crops, the potentical risk of their field release, the impact of soil mineral, organic matter and organo-mineral complex on the persistence of purified Bt toxins, and the behavior of Bt toxins in rhizospheric soil. Based on our researches, several problems which should be paid attention to in related studies were also pointed out. PMID- 12625013 TI - [Feasibility to introduce rare tree species Pinus sibirica into China]. AB - Pinus sibirica growing mainly in Siberia of Russia is distributed over the Euro Asia Taiga forest belt. There are many high-quality populations due to a great deal of variations. This kind of tree has an advantage of standing up to frigid environment, and can spread out in such places that have cold weather and high altitude. In China, boreal forest is a wide-spreaded type of forest that has the largest area and high volume. For this reason, it is feasible to introduce Pinus sibirica into the region that the condition is suitable. Introducing this kind of tree is a strategic project that can improve the structure and quality of our boreal forest. Introducing it can not only meet the demands of improved variety in short time, but also do the experiment of producing edible seeds and build up the developing center of nut, which can be a way of getting rid of poverty of forest region in heavy frigid area where is regarded as infertile area for farming formerly. PMID- 12625014 TI - [Research advances in plant selenium nutrition and its mechanism]. AB - The amount of selenium in soil environment significantly influences the selenium nutrition of plants. In this paper, the absorption, accumulation and translocation of Se in plants was discussed, and the toxicity of selenium and its effect on crop yield and quality was summarized. The advances in related researches were also introduced. On the basis of these, a new approach on the effect of selenium on soil enzyme activities to elaborate the essentiality of selenium for plants was proposed. PMID- 12625015 TI - [Stable mechanisms of soil aggregate and effects of human activities]. AB - Soil aggregate is the basic unit of soil structure. Its formation is a natural process but influenced severely by human activities, including land utilization, tillage disturbance, manure application, and cropping system. The improvement of soil structure is decided not only by total amount of organic C inputed into soil, but also by the composition and characteristic of organic matter. Owing to the different composition and characteristic of organic matter, there are different mechanisms for the stability of micro- and macro-aggregates. PMID- 12625016 TI - [Mechanisms of organic cementing soil aggregate formation and its theoretical models]. AB - Soil aggregate is the basic unit of soil structure, and its formation and stabilization has direct effect on soil fertility and crop growth. For black soil with high content of organic matter, organic cementing substance is an important factor in forming soil aggregates. Based on the recent researches on organic cementing agents, this paper was focused on the nature of organic cementing substances, the effect of soil organisms and plant roots in forming soil aggregates, and the theoretical models and stable mechanisms of soil aggregate formation. PMID- 12625017 TI - [Advances in research on ecological land classification]. AB - Land classification is the base of land evaluation, land planning and land management, and plays an important role in the sustainable development of forestry resource. Ecological land classification (ELC) is the main approuch and direction of land classification. Along with the development of landscape ecology and 3S techniques, developing of ecological classification system (ECS) becomes the keystone of ELC. The definition, history, basic characteristics of ELC, the theory of ECS and its' prospects were systematically reviewed in this paper. The prospects were also described for the development of ECS methodology and the application of ELC. The general direction of ecological land research, which was a multi-factors and multi-levels syntheses based on the multi-objects administration of resource (forest, land and waters), was the combination of quantificational and qualitative research. It is necessary to carry out relevant researches in China. PMID- 12625018 TI - [Unique ecosystem characters and ecological principles for development in the ecotones between agriculture and pasture in north China]. AB - Based on the fact that its ecological deterioration, a rethinking on the former definition of 'transitional zone between agriculture and grassland' was proposed. It was recognized that this zone has been gradually and dynamically formed under the specific circumstances of history and anthroponomy in China. It should thus be re-defined as 'ecotone between agriculture and pasture', instead of just focusing on its features of geography, ecology, and vegetation separately, as well as its character of transition. More attention should be put on its nature of complex system of ecology, society and economy, and then, derive relevant abating measures. Boreal ecotone has the functions of life support for a relatively dense population, as well as ecosystem services. Under the premise of unable copying the models for arid and semiarid areas in abroad, in particular developed countries, an unique strategy of capable of compromising both above mentioned functions, as well as ecological reconstruction, which is embodied in readjusting agro-structure, rationally utilizing and conserving resources, and establishing 'polar' industries, such as grasses and livestock, should be the center of an interdisciplinary study approach. PMID- 12625019 TI - [Advances in research on CO2 concentrating mechanism of green algae]. AB - Unicellular green algae plays a key role in freshwater ecosystem, which possesses a CO2 concentrating mechanism that can increase the level of CO2 at the active site of ribulose bisphosphate carboxylase-oxygenase (Rubisco) by actively transporting inorganic carbon when adapted to low CO2 concentration. The mechanism results in an increase in photosynthetic rate, and a decrease in photorespiration. This mechanism and its environmental regulation such as light, temperature, CO2 concentration and nutrient are reviewed in this paper to enhance further studies on response of phytoplankton to elevated atmospheric CO2 concentration in China. PMID- 12625020 TI - [A review on the concept, characteristic and assessment of landscape health]. AB - As a new concept emerging from landscape ecology and ecosystem health, landscape health is the subject dealing with ecosystem health for landscapes which, under the disturbance of intense activities from human beings, are seriously polluted and degraded, even facing with the risk of disappearing. The basic concept, characteristics and criteria of landscape health were introduced in this paper, and indicators, scales and hierarchical structures and methods for landscape health assessment were also discussed. It was emphasized that comparative, integrative, and developmental methods should be applied in landscape health assessment. PMID- 12625021 TI - [Preliminary study on the growth stimulation of marine microalgae stimulated by low level of toxicant]. AB - Cell growth of Nitzschia closterium and Platymonas subcordiformis was stimulated under 4 and 8 micrograms.L-1 anthrancene stress, respectively. The cell growth of two species was also stimulated under 30 mg.L-1 streptomycin stress. The hormesis of marine microalgae cells under low level of anthrancene and streptomycin resulted in low level of free radical and decrease of microorganisms. Furthermore, the hormesis of marine microalgae was also discussed in this paper. PMID- 12625022 TI - [Effect of waterlogging on senescence of flag leaf and root of wheat yangmai]. AB - In the pot culture experiment, winter wheat (Triticum aestivum L.) c.v. Yanmai 5 was selected to study the effect of long-term water-logging, which was conducted from jointing stage to 20 days after anthesis, on grain weight and membrane lipid peroxidation, and on enzyme activities involving membrane lipid peroxide elimination of wheat leaf and root. The content of MDA (malondialdehyde), the product of membrane lipid peroxidation in flag leaf and root under waterlogging increased significantly, while the activities of SOD (super oxide dismutase) and CAT (catalase), two key enzymes involving in membrane lipid peroxide elimination decreased severely. Thereafter, the wheat flag leaf and root under waterlogging senesced much earlier than those under control. Therefore, the grain filling rate of wheat grown under water-logging condition decreased significantly, and subsequently, its grain weight decreased obviously. PMID- 12625023 TI - New developments in neuroscience. PMID- 12625024 TI - Overview: the search for therapeutic strategies for cerebrovascular ischemia. PMID- 12625025 TI - Genome-based drug discovery: prioritizing disease-susceptibility/disease associated genes as novel drug targets for schizophrenia. AB - The search for new classes of antipsychotics based on novel targets identified from linkage/linkage association in diseased cohorts and microarray approaches using tissue from affected individuals is a high priority in central nervous system research. Genes linked to schizophrenia, a disease affecting 1% of the population, have been identified on nearly every chromosome of the human genome leading to a diverse choice of targets for validation. Interestingly, while the majority of currently used antipsychotic medications act by blocking dopamine receptors, there have been few genetic studies implicating the dopamine receptor family in disease etiology. Recently, four genes have been identified that encode dysbindin, neuroregulin, D-amino acid oxidase and G72, respectively, that support previous studies suggesting that schizophrenia may result from a hypofunction of glutamatergic neurotransmission. Linkage and microarray studies have similarly supported studies implicating the alpha 7 neuronal nicotinic receptor in the etiology of schizophrenia. Microarray studies using brain tissue from schizophrenic patients have shown changes in gene expression that number in the thousands, involving a number of proteins related to synaptic structure and function (PSYN gene group) and cellular metabolism. The majority of these proteins are not traditional drug discovery targets, nor are their functional roles in schizophrenia obvious, providing a challenge to validate them from the drug target identification/drug discovery perspective. The current state-of-the art in genome-based approaches to schizophrenia, target discovery highlights a need for a multidisciplinary, integrative, null hypothesis-based approach to sort through these novel genes as drug targets. PMID- 12625026 TI - New drug development for post-traumatic stress disorder. AB - US FDA approval of two serotonin-selective reuptake inhibitor (SSRI) agents for post-traumatic stress disorder (PTSD) has created new opportunities for drug development. This follows many years of exploring the potential utility of several classes of psychotropic agents for this very common, yet under-recognized and under-treated disorder. This review examines some of the basic neurobiological abnormalities observed in PTSD and summarizes open and controlled drug trials for major classes of medications, including SSRIs, other antidepressants, atypical neuroleptics, noradrenergic modulators and anticonvulsants, while critically evaluating the extent of effectiveness of these agents and reviewing unmet gaps in therapeutic need. PMID- 12625027 TI - New hope in the treatment of painful symptoms in depression. AB - Depression is increasingly seen as a triad of psychological, somatic and physical symptoms that all need to be treated to achieve maximal remission. In primary care, physical symptoms such as pain, are the principal presenting symptoms, and a common psychopharmacology between pain and depression suggests that compounds that inhibit the reuptake of both serotonin and norepinephrine are likely to produce the greatest relief from depression and chronic pain. Recent, principally open, trials with members of the new selective serotonin and norepinephrine reuptake inhibitor class of antidepressants such as venlafaxine, milnacipran and duloxetine (Eli Lilly & Co/Shionogi & Co Ltd), suggest that these compounds may be effective in relieving pain both associated with, and independent of depression. PMID- 12625028 TI - Corticotropin-releasing hormone modulators and depression. AB - Basic and clinical studies demonstrate that the central corticotropin-releasing hormone (CRH) circuits are overactive among depressives, a phenomenon frequently reflected by enhanced cortisol and corticotropin levels in the peripheral blood of these patients. Behavioral pharmacology provided evidence that CRH overexpression accounts for many signs and symptoms characteristic of depression. CRH-type 1 receptors (CRHR), were identified as responsible for conveying the CRH signal into cellular circuitries, thereby inducing depression-related symptoms. In order to decrease CRH signaling, many pharmaceutical companies have developed small molecules that after oral ingestion, penetrate the blood-brain barrier and selectively bind at CRHR1 with high affinity. These compounds have been tested in animal models and patients with major depression. One of these compounds, R 121919 (Neurocrine Biosciences Inc), ameliorated depressive symptomatology without unwanted endocrine side effects or other adverse effects. While clinical trials of R-121919 have been discontinued after phase IIa studies, a number of other CRHR1 antagonists are being developed, and hopefully this advance will ultimately lead to a favorable alternative to currently available antidepressant drugs. PMID- 12625029 TI - Therapeutic options for Huntington's disease. AB - Huntington's disease (HD) is an autosomal dominant, progressive neurodegenerative disorder. The genetic defect lies in the expansion of a CAG repeat on chromosome 4 and neuropathologically it is characterized by neuronal loss in the striatum. Clinical signs are chorea, impaired voluntary movement, behavioral changes and dementia. Present therapies are limited to these symptoms without any influence on the course of the disease, whereas current pharmacological developments mainly focus on delaying disease progression. This review discusses the present symptomatic treatments and focuses on recent developments of new therapeutic approaches for HD. PMID- 12625030 TI - Amyloid inhibitors and Alzheimer's disease. AB - Neuritic plaques composed of amyloid beta-protein (A beta) are an early and invariant neuropathological feature of Alzheimer's disease (AD). The current preclinical search for drugs is mainly focused on decreasing A beta production by inhibiting beta- or gamma-secretase, blocking the formation of these plaques by preventing A beta protofibril and fibril formation, and alleviating the toxic effects of neuritic plaque deposition. Increasing numbers of drugs currently used as therapies for other diseases are now entering clinical trials for AD, but the molecular targets of these drugs and their relevance to A beta toxicity needs to be thoroughly addressed. This knowledge will allow us to fully understand the A beta-related pathways in AD pathogenesis and explore novel therapeutic interventions. PMID- 12625031 TI - Tadalafil Lilly ICOS. AB - Tadalafil is a phosphodiesterase type 5 inhibitor in development by Lilly ICOS for the potential treatment of erectile dysfunction. The compound will be marketed in North America and Europe by a collaboration formed by Eli Lilly & Co and ICOS Corp. Eli Lilly & Co has marketing rights in all other territories. Marketing approval in Europe was granted in November 2002, with launch expected in the first half of 2003. An approvable letter was issued by the FDA in April 2002, with a US launch anticipated in the first half of 2003. PMID- 12625032 TI - RG-1068 RepliGen. AB - RG-1068 is a synthetic form of the natural human hormone secretin under development by RepliGen for the potential treatment of autism. RG-1068 received Fast Track designation for the treatment of pediatric autism in September 2001, and in February 2002, it entered phase III clinical trials. PMID- 12625033 TI - DU-127090 Solvay/H Lundbeck. AB - DU-127090 is a mixed dopamine antagonist/serotonin agonist in development by Solvay and H Lundbeck as a potential treatment for psychosis and schizophrenia, for which it is in phase II clinical trials. In August 2002, phase II trials were ongoing and Lundbeck expected to commence phase III trials in 2003 and file an NDA after 2004. DU-127090 is also under development for Parkinson's disease, for which it is in phase I clinical trials. PMID- 12625034 TI - Sumanirole Pharmacia. AB - Sumanirole is a dopamine D2 agonist under development by Pharmacia Corp (formerly Pharmacia & Upjohn) for the treatment of Parkinson's disease. The compound is currently undergoing phase II trials in Japan and phase III clinical trials in the US. PMID- 12625035 TI - Nicaraven Chugai. AB - Chugai (the Japanese subsidiary of Roche) is developing nicaraven (Antevas), a water-soluble antioxidant, for the potential treatment of disorders caused by acute cerebrovascular diseases. A registration application was filed in April 1995, and in April 2002, nicaraven was still awaiting registration in Japan. By August 2002, Chugai had filed an NDA in Japan for the additional indication of subarachnoidal bleeding. PMID- 12625036 TI - Pagoclone Indevus. AB - Pagoclone is a cyclopyrrolone GABAA receptor modulator under development by Indevus for the potential treatment of panic and anxiety disorders. Indevus initiated phase II/III trials in November 1996. Pfizer had initiated a phase III trial for panic disorder in August 2000 but discontinued development of the compound in June 2002. PMID- 12625037 TI - [Deceleration of vital activity as a universal biochemical mechanism ensuring adaptation of microorganisms to stress factors: a review]. AB - The review deals with a major problem of contemporary biology, i.e., the strategy of persistence of organisms affected by stressors. The primary mechanism of biochemical adaptation of cells is their transition to a specific surviving state, which is characterized by deceleration of vital activity (DVA), a process related to decreased metabolism. According to the latest classification, there are four DVA types: quiescence (endogenous or exogenous), cryptobiosis, anabiosys, and latent life. These states are viewed from a new standpoint, as depending on both the stage of growth and development of the organism and the type of stressor. A brief characterization of cryptobiosis and anabiosys is provided. Quiescence, which is a specific type of DVA, is characterized in greater detail, with emphasis on differences between fungi and bacteria. Endogenous and exogenous quiescence is compared in two types of cells (spores). Changes in the composition of quiescent cells (spores) as compared to their vegetating counterparts involve lipids (membrane-associated and neutral), cytosolic carbohydrates (which serve as a reserve supply of nutritives or act as protectors), water content, RNA/protein synthesis, and cyclic nucleotides (e.g., cAMP). Biochemical mechanisms controlling reversion of DVA (the transition from spores to germinating cells) are also analyzed. In conclusion, consideration is given to the role of DVA in evolution and to the implications of DVA studies for biotechnology, cryogenic preservation, utilization of interstellar space and permafrost territories. PMID- 12625038 TI - [Estimation of the hydrocarbon-oxidizing activity of microorganisms]. AB - A method allowing the microorganisms growing on substrates with a low water solubility (oil, fuel oil, resins, and asphalthenes) to be isolated and counted was developed. An advantage of the method is its ability to estimate the oil utilizing activity of each strain visually according to the decolorized zones formed during its growth on oil products. The sizes of these zones indicate which oil-degrading strain is most active. PMID- 12625039 TI - [Resistance of Penicillium piceum F-648 to hydrogen peroxide under short term and prolonged oxidative stress]. AB - Resistance of Penicillium piceum F-648 to hydrogen peroxide under short-term and prolonged oxidative stress was studied. An increase in the activity of intracellular catalase in fungal cells after short-term exposure to hydrogen peroxide was shown. Activation of fungal cells induced by H2O2 depends on H2O2 concentration, time of exposure, and the growth phase of the fungus. Variants of P. piceum F-648 that produced two forms of extracellular catalase with different catalytic properties were obtained due to prolonged adaptation to H2O2. Catalase with low affinity for substrate was produced predominantly by the parent culture and variant 3; however, a high substrate affinity of catalase was observed in variant 5. Variant 5 of P. peniceum F-648 displayed a high catalytic activity and operational stability of catalase in the presence of phosphate ions and the concentration of substrate less than 30 mM at pH more than 7. PMID- 12625040 TI - [Genetic aspects of the relationship between isoquinoline alkaloids and mineral elements in greater celandine (Chelidonium majus L.)]. AB - Interrelations between the total content of isoquinoline alkaloids, concentrations of quaternary protoberberines and benzophenanthridines, and the amount of K, Cu, Co, Al, Ba, and Zn in aerial parts of individual celandine plants were revealed, within a single cenopopulation, using correlation analysis and regression analysis. Mathematical models describing the regulation of isoquinoline metabolism by some of the mineral elements were obtained in the analytical form. The results suggest that this process is genetically determined. PMID- 12625041 TI - [Evaluation of the effects of biological preparations on phytopathogenic fungi Didymella applanata and Botrytis cinerea]. AB - Fungicidal and fungistatic effects of biological preparations involving bacteria of the genera Pseudomonas and Bacillus and fungi of the genus Chaetomium on phytopathogenic fungi Didymella applanata and Botrytis cinerea were evalauated. All the biological preparations under study inhibited the growth of colonies of the fungi; however, the degree of the inhibition depended on the nature of each particular microorganism and the concentration of each particular preparation. The preparation containing Bacillus subtilis at a concentration of 0.2% effected maximum suppression of B. cinerea (the diameter of the colonies decreased sevenfold). The preparations containing bacteria of the genus Pseudomonas and fungi of the genus Chaetomium were most efficient in suppressing D. applanata. The preparations containing B. subtilis and Chaetomium spp. showed promise as agents against simultaneous development of spur blight and Botrytis blight. PMID- 12625042 TI - [Conditions for cultivation of the fungus Penicillium melinii UzLM-4 and its biosynthesis of lipases]. AB - Cultivation of the fungus Penicillium melinii UzLM-4 on a Raistrick's medium of our own modification made it possible to increase the biosynthesis of lipases three to four times. The following conditions ensured a high rate of synthesis of the extracellular lipase: age of the inoculum, 15 days; concentration of the inoculum, 15 x 10(6) conidia per 100 ml medium; initial pH of the nutrient medium, 8.0; and cultivation in a shaker at 150 rpm (25 degrees C). PMID- 12625044 TI - [Effects of nitriles and amides on the growth and the nitrile hydratase activity of the Rhodococcus sp. strain gt1]. AB - Effects of some nitriles and amides, as well as glucose and ammonium, on the growth and the nitrile hydratase (EC 4.2.1.84) activity of the Rhodococcus sp. strain gt1 isolated from soil were studied. The activity of nitrile hydratase mainly depended on carbon and nitrogen supply to cells. The activity of nitrile hydratase was high in the presence of glucose and ammonium at medium concentrations and decreased at concentrations of glucose more than 0.3%. Saturated unsubstituted aliphatic nitriles and amides were found to be a good source of nitrogen and carbon. However, the presence of nitriles and amides in the medium was not absolutely necessary for the expression of the activity of nitrile hydratase isolated from the Rhodococcus sp. strain gt1. PMID- 12625043 TI - [Analysis of secondary metabolites of microscopic fungi of the genus Penicillium by chromatographic techniques]. AB - Combinations of various regimens of thin-layer chromatography and high performance liquid chromatography (HPLC) was efficient in analyzing 39 nitrogen containing secondary metabolites (alkaloids) produced by 12 strains of microscopic fungi of the genus Penicillium. Chromatographic mobility of alkaloids on Silufol plates was determined in the following systems (following staining with the Ehrlich reagent): (a) chloroform, methanol, and 25% NaOH (90:10:1, 90:10:0.1, or 80:20:0.2); (b) chloroform and acetone (9:1); and (c) ethyl acetate, methanol, and 25% NH4OH (85:15:10). Conditions for separation of clavine alkaloids by HPLC on Spherisorb ODS2 and Supelcosil LC-18 columns (gradient elution) were optimized. Retention values of 22 alkaloids were compared to those of agroclavine and roquefortine. PMID- 12625045 TI - [Composition and biological activity of submerged mycelium of the xylotrophic basidiomycete Lentinus edodes]. AB - The composition of submerged mycelium of Lentinus edodes, grown in laboratory fermenters, has been studied. The mycelium contained 23-24% proteins, 8-9% lipids, up to 1800 mg% phenolic substances, and a significant amount of inorganic substances, including calcium and iron. The fungus produced up to 5.0% intracellular and 3.5-4.0 g/l extracellular polysaccharides. The submerged mycelium stimulated the development of humoral immune response elicited by sheep red blood cells. PMID- 12625046 TI - [Fungal decomposition of oat straw during liquid and solid state fermentation]. AB - White rot fungi (Coriolus hirsutus, Coriolus zonatus, and Cerrena maxima from the collection of the Komarov Botanical Institute of the Russian Academy of Sciences) and filamentous fungi (Mycelia sterilia INBI 2-26 and Trichoderma reesei 6/16) were grown on oat straw-based liquid and solid media, as well as in a bench-scale reactor, either individually or as co-cultures. All fungi grew well on solid agar medium supplemented with powdered oat straw as the sole carbon source. Under these conditions, the mould Trichoderma reesei fully suppressed the growth of all basidiomycetes studied; conversely, Mycelia sterilia neither affected the development of any of the cultures, nor did it show any substantial susceptibility to suppression by their presence. Pure solid cultures of basidiomycetes, as well as the co-culture of Coriolus hirsutus and Cerrena maxima caused a notable bleaching of the oat straw during its consumption. When grown on the surface of oat straw-based liquid medium, the basidiomycetes consumed up to 40% polysaccharides without measurable lignin degradation (a concomitant process). Under these conditions, Mycelia sterilia decomposed no more than 25% lignin in 60 days, but this was observed only after polysaccharide exhaustion and biomass accumulation. In contrast, during solid state straw fermentation, white rot fungi consumed up to 75% cellulose and 55% lignin in 83 days (C. zonarus), whereas the corresponding consumption levels for co-cultures of Mycelia sterilia and Trichoderma reesei equaled 70 and 45%, respectively (total loss of dry weight ranged from 55 to 60%). Carbon dioxide-monitored solid-state fermentation of oat straw by the co-culture of filamentous fungi was successfully performed in an aerated bench-scale reactor. PMID- 12625047 TI - [Comparative analysis of models describing interactions between antibodies and liposomal antigens]. AB - Mathematical models of three levels of complexity are proposed, describing the interactions between antibodies and polyvalent liposomal antigens. The models take into account the contribution of high-affinity bivalent complexes and clusters of several antigenic groups to the immunochemical reaction. The models were analyzed numerically at different values of thermodynamic and kinetic parameters of the antigen-antibody reaction. Conditions, under which models of different levels of complexity provide a satisfactory description of the antigen antibody interaction, were determined. The adequacy of the results of modeling was confirmed experimentally in liposomal preparations differing from each other by the surface density of monovalent hapten (atrazine) conjugated with a lipid. PMID- 12625048 TI - [Biological oxidation of sulfide raw material using a culture of Thiobacillus ferrooxidans under various conditions of leaching]. AB - Major parameters of the first stage of leaching of a copper-zinc sulfide product by a culture of Thiobacillus ferrooxidans have been studied, including the effects of solid phase concentration, Fe2+ and Fe3+ ions, pH, and the intensity of mixing. The first stage of leaching of the sulfide raw material is optimum under the following conditions: pH of the original leaching solution equal to 1.6; Fe3+ concentration of order of 10 g/l; and vigorous mixing of the suspension at solid phase concentrations of 30-35%. A theoretical substantiation of the observed dependences is proposed. PMID- 12625049 TI - [Determination of carboxylic acids in wines by liquid ion-exclusion chromatography]. AB - Chromatographic behavior of aromatic and aliphatic acids was studied using an ion exclusive column. Conditions were determined, under which the separation of the acids by single-column ion-exclusion chromatography with UV-detection was the most selective and efficient. The method developed by us was employed for determining the content of carboxylic acids in wines. PMID- 12625050 TI - [Effect of the structure of alcohols on their binding by thermotropic gels of corn starch]. AB - Binding of alcohols by thermotropic gels of normal corn starch was studied using the method of capillary gas-liquid chromatography. The amount of the gel-bound substances depended linearly on their concentrations in the original gel. Binding of n-alcanols increased with the length of the alkyl chain, exhibiting a linear dependence. The binding was attenuated by the presence of double bonds-cyclic substituents in or branching of the carbon chain. Binding of alcohols by starch polysaccharides was more pronounced in thermotropic gels, as compared to cryotextured gels. Binding isotherms were indicatice of co-operative interactions, in which alcohols formed supromolecular structures with hydrated molecules of polysaccharides and inclusion complexes with amylose and side chains of amylopectin. PMID- 12625051 TI - [Studies of oligosaccharide specificity of perch roe fucolectin, using gold labeled neoglycoproteins]. AB - The specificity of perch (Perca fluviatilis) roe fucolectin was studied using the protein dot blot technique, followed by detection with colloidal gold-labeled neoglycoproteins bearing human milk polysaccharides. The strongest binding was noted with the H type 1 pentasaccharide lacto-N-fucopentaose (LNFP I, Fuc alpha 1 2 Gal beta 1-3 GlcNAc beta 1-3 Gal beta 1-4Glc); the interaction with the H type 6 trisaccharide 2'-fucosyllactose (2-FL, Fuc alpha 1-2 Gal beta 1-4 Glc) was weaker. Binding of the perch lectin to the Lewis antigens (associated with tumors and embryonic tissues) was also studied. It was found that the lectin weakly interacted with the hexasaccharide lacto-N-difucohexaose I, Fuc alpha 1-2 Gal beta 1-3[Fuc alpha 1-4]GlcNac beta 1-3 Gal beta 1-4 Glc), but not with Lea, Lec, Lex antigens. Thus, perch roe lectin exhibited pronounced differences in carbohydrate specificity from other fucolectins--a feature that may be used in structural studies and isolation of fucose-containing glycoconjugates. PMID- 12625052 TI - [Role of trophic and hormonal factors in exogenous regulation of the formation of reproductive organs in yellow lupine (Lupinus luteus L.)]. AB - During the formation and ripening of beans in yellow lupine, their intactness may be increased by treatment of the plants with the microelement molybdenum or the growth factor 24-epibrassinolide. Early stages of the effects of either factor involved activation of various metabolic pathways; eventually, the supply of fruit elements with free amino acids was improved and the seed productivity of the plants, increased. PMID- 12625053 TI - [Specific features of albumin interactions with hemiacetals of aflatoxins and sterigmatocystin]. AB - Aflatoxin B2a (AB2a), aflatoxin G2a (AG2a), and hemiacetal of sterigmatocystin have been shown to form immunoreactive conjugates with albumin. The conjugates were formed following incubation of solution mixtures at room temperature for 1 h, as demonstrated by spectrophotometry and enzyme immunoassay. Anti-AB2a antibodies reacted with AB2a, aflatoxin B1, and aflatoxin AB2 (100, 8.8, and 5.9%, respectively); a similar result was obtained for anti-AG2a antibodies reacting with AG2a, aflatoxin G1, and aflatoxin AG@2 (100, 2.5, and < 1.0%, respectively). Binding of anti-AB2a and anti-AG2a antibodies to solid-phase conjugates of AB2a or AG2a exhibited similar analytical characteristics. PMID- 12625054 TI - [Primary structure of a short toxin from the venom of a vietnamese scorpion (Buthus sp.)]. AB - The complete amino acid sequence of an important toxin (toxin 14) from the venom of a Vietnamese scorpion (Buthus occitanus sp.) has been determined, which includes 35 amino acid residues and three disulfide bridges (molecular weight, 3843 Da). The comparison of the sequence with sequences of short scorpion toxins led us to conclude that toxin 14 belongs to a novel group of toxins affecting the excitability of myelinated nerves. PMID- 12625055 TI - [Iurii Viktorovich Natochin ( on the 70th anniversary of his birth)]. PMID- 12625056 TI - [The role of beta,gamma-dimers of GTP-binding proteins in the processes of hormone signal transduction]. PMID- 12625057 TI - [Neurochemical organization of the turtle pretectum: immunohistochemical study. Comparative analysis]. PMID- 12625058 TI - [New methods to analyze the variations of the brain gangliosides content, which are connected with idioadaptation and aromorphosis in vertebrates]. PMID- 12625059 TI - [Distribution of the proteins, providing for the iron ions homeostasis in the cow eye retina ]. PMID- 12625060 TI - [The complex study of the Drosophila melanogaster mutants in agnostic locus: the model for the disruption of the genome architecture and cognitive functions]. PMID- 12625061 TI - [The study of the prostaglandins influence on the water intake and ion transport in the skin of the frog Rana temporaria]. PMID- 12625062 TI - [The study of the role of the GABA A and GABA B receptors in presynaptic inhibition of primary afferents of spinal cord of the frog Rana ridibunda]. PMID- 12625063 TI - [Wulst neurons in the youngs of pied flycatcher Ficedula hypoleuca at previsual stage of development]. PMID- 12625064 TI - [Comparative study of the relaxation mechanism in the smooth muscles of bronchi and pulmonary artery in rat]. PMID- 12625065 TI - [Morphological study of the space distribution of vestibulospinal neurons in the frog Rana ridibunda]. PMID- 12625066 TI - [Catecholamines and nitric oxide participation in the regulation of apoptosis in nonapeptidergic neurons in the rat hypothalamus]. PMID- 12625067 TI - [XVIII Russian Physiology Congress ]. PMID- 12625068 TI - [History of modern chromosomal analysis. Differential staining of plant chromosomes]. AB - Differential staining methods found extensive use in karyotype studies of many plant and animal species and provide for reliable identification of all chromosomes of the organism. Below we describe the most widespread methods and history of their advent. In addition, we discuss specific structure of the chromosomes and possible mechanisms responsible for differential segmentation. PMID- 12625069 TI - [Aspartate- and alanine-aminotransferase in early development of keta]. AB - We studied the activities of the marker enzymes of physiological state and adaptive reactions, aspartate and alanine aminotransferases, in early development of the keta Oncorhynchus keta. Aspartate aminotransferase with pH optima 6.8, 7.0, 7.6, and 8.0 and alanine aminotransferase with pH optima 7.0, 7.4, 7.6, 7.8, 8.0, and 8.2 were found in the eggs, larvae, and fry. The succession of enzymes with different pH takes place during ontogenesis, as well as stage specific changes in their activity. The maximum enzymatic activity was recorded in the larvae during their rise for "afloat." A correlation was established between the dynamics of enzymatic activity and soluble nitrogen and amine nitrogen contents. PMID- 12625070 TI - [Age-related features of cataractogenesis in salmon fry. I. Lipid composition of the lens in normal development]. AB - This papers opens a series of publications on the mechanisms of cataractogenesis in the salmon fry. Biochemical features of normal lens development and cataractogenesis in fry of different age and age-related dynamics of the liver lipid composition during upon cataractogenesis will be dealt with, since lipids are most intensely synthesized in the liver, from where they are transported in the lens. Here, we describe the dynamics of lens lipid composition in the salmon fry, including the total lipid content and dynamics of individual classes. The data are analyzed on the fatty acid spectrum of the salmon fry lens, as compared to the human lens. PMID- 12625071 TI - [Hematopoiesis on cellulose acetate membranes in the presence of lipopolysaccharides]. AB - The formation of hemopoietic colonies on acetate cellulose membranes in the peritoneal cavity of mice was markedly enhanced after the injection of bacterial lipopolysaccharide. In addition to granulocytic-macrophagal differentiation, the foci of erythropoiesis appeared. The stimulating effect of lipopolysaccharide was not expressed in nonirradiated mice and during the formation of hemopoietic foci on acetate cellulose membranes in the subcutaneous connective tissue. PMID- 12625072 TI - [Effect of chorionic gonadotropin on thymocyte differentiation in the presence of thymus epithelial cells]. AB - We studied the effects of the main placental hormone, chorionic gonadotropin, on differentiation of human thymocytes in vitro in the presence of thymic epithelial cells. It was shown that the hormone at a high dose (100 IU/ml) enhanced the epithelium-induced phenotypic maturation of thymocytes, which is registered by an increased expression of the membrane marker CD3 and transition of CD4+8+ thymocytes in the cells with CD4+8- and CD4-8+ phenotypes. In addition, gonadotropin enhanced the proliferative response of thymocytes to the mitogen during their cultivation with the epithelium. The stimulating effect of the hormone on the epithelium-induced differentiation of thymocytes is mediated by the humoral factors of epithelial cells. In addition, gonadotropin at this dose exerts its own differentiating activity with respect to thymocytes and stimulates their phenotypic and functional maturation in a monoculture. PMID- 12625073 TI - [Regulation of asexual reproduction in the planarian Dugesia tigrina]. AB - We studied asexual reproduction of planarians under the natural and artificial photoperiodic conditions. It was shown that light inhibits the fission of planarians, while darkness stimulates it. The diurnal dynamics of the fission of planarians demonstrated a circadian rhythm. This rhythm is stable, which is expressed when the conditions are experimentally changed: constant darkness, unnatural rhythm of light-darkness succession). However, this stability is affected at the time zone change. The planarians are adapted to new conditions and begin to fission at once in correspondence with the new diurnal regime. PMID- 12625074 TI - [Effect of juvenoids and retinoic acid on the development or larva and nymphs of the tick Ixodes ricinus L. (Acari: Ixodidae) and regeneration of Haller's organ during metamorphosis]. AB - Larvae and nymphae of the tick Ixodes ricinus L. display similar reactions to analogs of the insect juvenile hormones (methopren and pyriproxyfen), which induce at both stages juvenalization of the Haller's sense organ regenerates. Similar effects were also described for retinoic acid. Unlike juvenoids, retinoic acid can affect not only regeneration, but also normal development of the Haller's organ and cause changes corresponding to so-called regenerative induction. Amputation of the leg and treatment with retinoic acid do not affect the duration of larval or nymphal development, while juvenoids somewhat accelerate their development. PMID- 12625075 TI - [Character of distribution of defects of wing venation in the banded agrion (Calopteryx spendens)]. AB - The distribution of morphological structures was studied in wings of the banded agrion (Calopteryx splendens Harr.) from different intrapopulation groups. Dragon flies of odd years of emergence are characterized by a more stable pattern of ontogenetic processes accordin to the mean total number of venation defects. The sharply increased level of radiation in summer 1986, which coincided with the flight of dragon-flies, could have caused hereditary defects expressed in a sequence of generations of even years of emergence. Apparently, most alternative features of wing venation in dragon-flies can be considered as markers of stability of the ontogenetic processes, which reflect, to a great extent, genotypic features of the organisms in a population. A possible mechanism has been described, which explains the proposed topological model of formation of the venation defects of different types. The increase in mean frequencies of defects can suggest an enhanced development over the aberrant epigenetic trajectories, which may lead to the elimination of these organisms under the influence of various agents, i.e., to the stabilizing selection in a population. The results obtained suggest that defects of venation arise on the stochastic basis and their frequency increases upon destabilization of ontogenetic processes not only by the environmental factors, but also by genetic stress. Venation defects can be successfully used in population biomonitoring. PMID- 12625076 TI - "I've been hit". PMID- 12625077 TI - Yuppie scans from head to toe: unethical entrepreneurism. AB - Our responsibility as Canadian radiologists is the same as the responsibility of physicians irrespective of the health care system they serve or the country where they work. "Our modern society does not value properly the intellectual effort, and the time studying the problems of patients and deciding on their solutions. Much higher rewards are placed on the use of sophisticated medical technology. Changing this way of practice requires much more than an academic call to order. As members of our modern society we physicians have a tough time isolating ourselves from prevalent values. But, we do have the duty to point out inconsistencies of the present societal values regarding medical activities, striving to change this state of affairs for the benefits of medicine, patients and society as a whole." PMID- 12625078 TI - Rational computed tomography screening in 2003. PMID- 12625079 TI - Interpretation of diagnostic tests: diagnostic accuracy efficacy and effectiveness. PMID- 12625081 TI - Patient positioning after lung biopsy: influence on the incidence of pneumothorax. AB - OBJECTIVE: To determine if post-biopsy complication rates were influenced by patient positioning after the procedure. METHODS: A prospective evaluation of post-biopsy complications in 87 patients (55 men, 32 women; mean age 66 yr, range 20-86 yr) undergoing fine-needle aspiration biopsy of lung nodules was performed. Biopsies were performed under computed tomographic guidance in 66 patients, fluoroscopic guidance in 18 and ultrasonographic guidance in 3 patients. Patients were randomly assigned to be placed either in the lateral decubitus "biopsy-side down" position or in the "biopsy-site-dependent" position (i.e., lying directly on the skin puncture site) after surgery. Chest imaging was performed immediately and at 4 hours after biopsy. RESULTS: Immediately after the procedure, 6 (14%) of the 42 patients in the decubitus biopsy-side-down group and 15 (33%) of the 45 patients in the site-dependent group experienced pneumothoraces (p > 0.05). No patient required tube thoracostomy. Two patients in the decubitus group and 1 in the site-dependent group experienced hemoptysis immediately after biopsy. On the 4-hour chest radiographs, there was no progression in size of the existing pneumothoraces. CONCLUSION: Positioning the patient either decubitus biopsy-side down or lying directly on the puncture site after lung biopsy does not appear to affect complication rates. PMID- 12625080 TI - Prediction of anemia on unenhanced computed tomography of the thorax. AB - OBJECTIVE: To determine if anemia can be predicted on unenhanced computed tomography (CT) of the thorax. METHODS: Hemoglobin and hematocrit levels were obtained within 24 hours of the unenhanced CT scan of the thorax of 200 patients. Anemia was defined as a hemoglobin level less than 140 g/L for men and less than 120 g/L for women. Regions of interest were placed on the left ventricular cavity, aorta and the interventricular septum if visualized. The attenuation of the interventricular septum and left ventricular cavity were correlated with the presence or absence of anemia. RESULTS: When the interventricular septum was not visualized, for every 1 Hounsfield unit (HU) increase in left ventricular attenuation, hemoglobin increased by 0.435 g/L (SE = 0.253, p < 0.001). Failure to visualize the interventricular septum did not exclude the presence of anemia in either sex. When the interventricular septum was visualized, 100% of males and 89% of females met the criteria for the diagnosis of anemia. The prediction of anemia by visualization of the interventricular septum alone yielded a sensitivity of 75.4% and a specificity of 90.3%, with 80% of patients correctly predicted. The multiple regression analysis model yielded a sensitivity of 94.2% and a specificity of 67.7%, with 86% of patients correctly predicted. CONCLUSION: The diagnosis of anemia should be suggested whenever the interventricular septum is visualized on unenhanced CT. PMID- 12625082 TI - Globus sensation: findings on videofluoroscopic examinations. AB - OBJECTIVE: To determine the findings on videofluoroscopy of the pharynx and esophagus in patients with a globus sensation and compare them with a similar group of patients with dysphagia. METHODS: A computerized search of radiology files revealed 83 patients with a globus sensation who had videofluoroscopic examinations of the pharynx and esophagus during a 10-year period from 1989-1999. A computerized search was also performed to generate an age- and sex-matched group of 83 patients with dysphagia who also had videofluoroscopic examinations during this period. Original radiologic reports were reviewed to determine the frequency of morphologic and functional abnormalities of the pharynx and esophagus in these 2 groups. RESULTS: Early closure of the cricopharyngeus was found in 13 (16%) patients with globus versus 2 (2%) with dysphagia (p < 0.01). Enlarged pharyngeal tonsils were found in 31 (37%) patients with globus versus 17 (21%) with dysphagia (p < 0.05). Enlarged lingual tonsils were characterized on barium studies by nodularity or lobulation of the posterior-inferior base of the tongue and valleculae in all patients, and the palatine tonsils were symmetrically enlarged in all but 2 patients. No other radiographic findings were significantly associated with a globus sensation. CONCLUSION: Early closure of the cricopharyngeus and enlarged lingual or palatine tonsils may contribute to the development of a globus sensation, but the pathophysiologic basis for this association remains unclear. PMID- 12625083 TI - Radiologic features of pancreatic lipoma. PMID- 12625085 TI - Experience with a modified Gray-St. Louis catheter for percutaneous gastrojejunostomy. AB - PURPOSE: To determine if design modifications to the Gray-St. Louis gastrojejunostomy catheter would improve its deployment and utilization characteristics. METHODS: A modified catheter and the Gray-St. Louis catheter were alternately inserted in consecutive patients requiring gastrojejunostomy. At the time of insertion, the anatomic locations of the fenestrated end of the catheter and the Cope retention loop were recorded. At the time of removal or exchange, catheters were assessed for the position of the fenestrated end and the retention loop, the presence of catheter tears, leakage of injected contrast medium and evidence of catheter blockage. RESULTS: Of the 250 catheters we assessed in a 3-year period, 141 (56%) were modified catheters and 109 (44%) were Gray-St. Louis catheters. At insertion, the retention loop of the modified catheter was in the duodenum in 132 (94%) patients, whereas the retention loop of the Gray-St. Louis Catheter reached the duodenum in 29 (27%) (p < 0.05). At the time of catheter removal or exchange, the retention loop of the modified catheter was in the duodenum in 72 (60%) patients, whereas the retention loop of the Gray St. Louis catheter was in the duodenum in 13 (14%) (p < 0.05). CONCLUSION: The design modifications to the Gray-St. Louis catheter improved the likelihood of the retention loop residing in the duodenum, while continuing to allow the fenestrated portion of the catheter to reside in the jejunum. PMID- 12625084 TI - Abdominal computed tomographic findings of Mycobacterium tuberculosis and Mycobacterium avium intracellulare infection in HIV seropositive patients. AB - OBJECTIVE: To compare the computed tomographic (CT) findings of abdominal Mycobacterium tuberculosis (MTB) infection and Mycobacterium avium intracellulare (MAI) infection in patients with human immunodeficiency virus (HIV) infection. METHODS: A retrospective review of the CT findings of 30 patients with HIV and proven MTB (n = 9) or MAI (n = 21) infection was conducted. Images were reviewed by a radiologist blinded to the diagnosis, and the radiologic findings involving the abdominal viscera, peritoneum and lymph nodes were compared. RESULTS: The following were more frequent in patients with MAI infection: hepatomegaly (MAI 71% v. MTB 44%, p < 0.05), uniform attenuation of lymph nodes (MAI 90% v. MTB 55%, p < 0.05) and clustered pattern of lymph nodes (MAI 57% v. MTB 22%, p < 0.05). In patients with MTB infection, lymph nodes with low attenuation centrally were more common (MAI 10% v. MTB 44%, p < 0.05), and mesenteric lymph nodes were significantly larger (MAI mean = 20 mm v. MTB mean = 40 mm, p < 0.05). CONCLUSION: Although nonspecific, CT may be useful in the early diagnosis of MTB and MAI infection, allowing for presumptive treatment before microbiologic confirmation. PMID- 12625086 TI - Fetal echocardiography--beyond the four-chamber view into the next millennium. PMID- 12625087 TI - Empathy: Its ultimate and proximate bases. AB - There is disagreement in the literature about the exact nature of the phenomenon of empathy. There are emotional, cognitive, and conditioning views, applying in varying degrees across species. An adequate description of the ultimate and proximate mechanism can integrate these views. Proximately, the perception of an object's state activates the subject's corresponding representations, which in turn activate somatic and autonomic responses. This mechanism supports basic behaviors (e.g., alarm, social facilitation, vicariousness of emotions, mother infant responsiveness, and the modeling of competitors and predators) that are crucial for the reproductive success of animals living in groups. The Perception Action Model (PAM), together with an understanding of how representations change with experience, can explain the major empirical effects in the literature (similarity, familiarity, past experience, explicit teaching, and salience). It can also predict a variety of empathy disorders. The interaction between the PAM and prefrontal functioning can also explain different levels of empathy across species and age groups. This view can advance our evolutionary understanding of empathy beyond inclusive fitness and reciprocal altruism and can explain different levels of empathy across individuals, species, stages of development, and situations. PMID- 12625089 TI - Adolescent health--a new perspective in Singapore. PMID- 12625088 TI - Two visual systems and two theories of perception: An attempt to reconcile the constructivist and ecological approaches. AB - The two contrasting theoretical approaches to visual perception, the constructivist and the ecological, are briefly presented and illustrated through their analyses of space and size perception. Earlier calls for their reconciliation and unification are reviewed. Neurophysiological, neuropsychological, and psychophysical evidence for the existence of two quite distinct visual systems, the ventral and the dorsal, is presented. These two perceptual systems differ in their functions; the ventral system's central function is that of identification, while the dorsal system is mainly engaged in the visual control of motor behavior. The strong parallels between the ecological approach and the functioning of the dorsal system, and between the constructivist approach and the functioning of the ventral system are noted. It is also shown that the experimental paradigms used by the proponents of these two approaches match the functions of the respective visual systems. A dual-process approach to visual perception emerges from this analysis, with the ecological-dorsal process transpiring mainly without conscious awareness, while the constructivist-ventral process is normally conscious. Some implications of this dual-process approach to visual-perceptual phenomena are presented, with emphasis on space perception. PMID- 12625090 TI - The perils of puberty. AB - Puberty and adolescence represent a transitional phase from childhood to adulthood, and are defined by the attainment of secondary sexual characteristics and reproductive capability, occurring concomitantly with psychological and social development. The perils of puberty lie in the difficult and complex inter relationship of biological, psychological and social changes, as the adolescent strives to adjust to new social roles and identities, and new reproductive capacities. An appreciation of these changes in normal puberty is essential to all professionals caring for adolescents. PMID- 12625091 TI - Five-year review of adolescent mental health usage in Singapore. AB - INTRODUCTION: To provide a review of adolescent mental health service utilisation in a child and adolescent mental health outpatient clinic in Singapore. MATERIALS AND METHODS: Data from all new adolescent cases seen over a 5-year period in the Child Guidance Clinic were analysed. A review of services provided is also included. RESULTS: Adolescent mental health usage has been on an increase with a rise in the number of cases diagnosed with depression. There has also been an increase in the number of forensic cases seen in the department. CONCLUSIONS: There is a growing demand for adolescent mental health services. As the demands and stresses on today's youth increase, there is likely to be a continued increase in the demand for such services. The challenge in the future is to provide adolescent mental health services in cost-effective packages that will meet the needs as well as stay financially viable. PMID- 12625092 TI - Youth suicide and parasuicide in Singapore. AB - INTRODUCTION: Suicide and parasuicide rates are important markers of the health of a nation. Suicide is a common cause of mortality in youth and parasuicide has its peak prevalence in adolescence and early adulthood. Both pose a tremendous burden to individuals, families and society. Historical and current studies on suicide and parasuicide in Singapore are reviewed and discussed in the light of available worldwide research. METHODS: Studies on suicide and parasuicide in Singapore were identified through a MEDLINE search (from 1979). Information from these studies were supplemented with relevant local monographs on suicide and parasuicide. RESULTS: Suicide is extremely rare in children under 10 years of age. Rates rise in the 10 to 14 years age group and increase markedly in the 15 to 19 years age group. The male to female ratio is about 2:1 for youths 10 to 14 years old and 1:1 for the 15 to 19 years age group. The historical preponderance of female suicide is much less clearly seen in recent years. Jumping from a height is the commonest method used, and mental illness and recent life stress are common causative factors. A peak in suicides in the young was found for June and October, and a trough for November and December. Parasuicide is extremely rare in children under 10 years of age. There was a peak in the months of October and November in the student population, corresponding to the examination months. Youths of Indian ethnicity were over-represented. CONCLUSIONS: Preventing suicide and parasuicide is of paramount importance, given the severe burden of such acts on individuals, families and society. Current studies on youth suicide in Singapore and a national study on parasuicide are urgently needed to increase our understanding of these phenomena. PMID- 12625093 TI - Eating disorders in Singapore: a review. AB - INTRODUCTION: Anorexia nervosa, bulimia and binge-eating disorder are the three best recognised eating disorders. Eating disorders have been reported to affect some 1 to 3% of adolescent females in the West. Increasing reports from Asia suggest that it is no longer possible to view these illnesses as 'Western' diseases. Eating disorders are the third most common serious medical disorder in female adolescents, after asthma and depression. It poses a tremendous burden for individuals, families and society. Historical and current studies on eating disorders in Singapore are reviewed, and discussed in the light of available regional and worldwide research. METHODS: Studies on anorexia nervosa and bulimia in Singapore were identified through a MEDLINE search (from 1979). Information from these studies were supplemented with relevant local monographs on eating disorders. RESULTS: Reports of eating disorders have increased over recent years in the local literature. The psychopathology of eating disorders in Singapore is very similar to that described in the Western literature, and in the two main current classification systems. Body dissatisfaction is prevalent among Singaporean Chinese schoolgirls and female undergraduates, with rates not dissimilar to the West. There is a link between 'Westernisation' (based on English being spoken as the main language at home) and body dissatisfaction. PMID- 12625094 TI - Sexually transmitted infections in Singapore teenagers. AB - INTRODUCTION: This is a review of sexual knowledge, behaviour and the extent of sexually transmitted infections (STI), including human immunodeficiency virus (HIV) infection, in Singapore adolescents. METHODS: Data were retrieved from relevant studies, reports and statistics concerning STI and HIV in Singapore. RESULTS: For many reasons, adolescents are at a higher risk of acquiring STI and HIV infections. Adolescents in Singapore are still relatively conservative in their sexual attitudes and behaviour. Most of those who had sex never used condoms. Fortunately, the incidence of STI and HIV infections among adolescents is relatively low. However, this may be changing, and there is a proportion of individuals who exhibit higher risk behaviours. CONCLUSIONS: In this age of rapidly changing morals and values, accurate information and skills need to be provided to young people to ensure that they are adequately prepared to protect themselves from acquiring STI/HIV infection. PMID- 12625095 TI - Adolescent alcohol and other substance use: sharing the Australian experience. AB - Substance use is common among adolescents. Although substance use may be a manifestation of experimentation in adolescent development, it may have more serious implications. It may be an indicator of emotional disturbance, or may be associated with several other health risk behaviours in adolescence. Substance use may also progress to harmful levels that can have a significant impact on a young person's health. Health professionals are increasingly being exposed to teenagers who use alcohol and drugs. The following article aims to provide, based on clinical and research experience in a number of institutions in Melbourne, Australia, an overview of the epidemiology of adolescent drug use, a clinical approach to assess substance use in young people and the principles of management of drug use in adolescence. PMID- 12625096 TI - Strategies to promote better outcomes in young people with chronic illnesses. AB - INTRODUCTION: This paper aims to provide an overview of chronic illness and disability in young people, focusing on the developmental needs of young people and the impact of chronic illness on adolescence. The emotional well-being of the young person with chronic illness is explored and strategies that promote better health outcomes for the young person through health, family and school settings are discussed. METHODS: Literature was searched using Medline from 1980 to 2002 using the keywords chronic illness, chronic disease, disability, psychological, social, outcomes and transition. Articles were also hand searched from retrieved articles. CONCLUSIONS: Young people with chronic illness have the same developmental needs as other adolescents. Traditional health care is focused on disease outcomes; however, young people do not necessarily share the same focus. Improving health outcomes for young people involves health professionals having an understanding of adolescent development and broadening the medical perspective to incorporate the developmental perspective. Building competence in young people with chronic illness, strengthening connectedness within the family, school and peer group and building the capacity of the health care team in different settings will provide opportunities to enhance resilience in young people with chronic illness. In doing so, their health outcomes will hopefully also be improved. PMID- 12625098 TI - Clinical assessment, management and outcomes of a group of adolescents presenting with complex medico-psychosocial conditions. AB - BACKGROUND: Adolescents with complex medico-psychosocial presentations are often seen as a management challenge. The Medical Family Therapy model provides a useful framework for working with these patients in the context of a multidisciplinary approach to treatment. MATERIALS AND METHODS: A retrospective case analysis of 38 patients referred over a two-year period to the Department of Adolescent Medicine was carried out. These patients met DSM-IV criteria for somatoform disorder or had a diagnosis of chronic fatigue syndrome (CFS). Duration of symptomatology, diagnosis, the presence of psychiatric conditions in the young person and their immediate family and the type and duration of the intervention were examined in relation to outcome. Two case presentations illustrate the complexity of the assessment and treatment process. RESULTS: Clinicians rated 47% of patients who engaged with the service as improved. There was no relationship between diagnosis, length of intervention and outcome. No significant differences emerged between the group of young people diagnosed with CFS and those with somatoform disorders in terms of outcome. Nine patients presented with symptoms which were similar or identical to those of one of their parents. Physical illness was more likely to be reported as a precipitating factor in the CFS group. Poor school attendance and psychiatric morbidity were linked to poor outcome. CONCLUSIONS: A comprehensive evaluation of presenting symptomatology and focussed intervention with measurable outcomes are important aspects of the clinical approach to complex medico-psychosocial conditions in adolescents. Families' beliefs about the presenting symptomatology and experiences of illness should be explored. PMID- 12625097 TI - Adolescent chronic illness: a qualitative study of psychosocial adjustment. AB - INTRODUCTION: The purpose of this study was to investigate the psychosocial issues facing young people living with a chronic medical condition. MATERIALS AND METHODS: Subjects were young people with a range of medical conditions who were on a waiting list to participate in the Chronic Illness Peer Support programme at the Centre for Adolescent Health, Royal Children's Hospital, Melbourne, Australia. Young people agreed to in-depth interviews which were taped and transcribed. Thematic analysis was undertaken by two researchers working independently. RESULTS: Thirty-five young people were interviewed. Thematic analysis revealed five broad themes: control (in control, under control, out of control); emotional reactions (happiness, frustration, anger, sadness, anxiety); acceptance (of illness, of others, of self); coping strategies, and; a search for meaning. The importance of social connections was emphasised. While illustrating the difficulties of managing a chronic medical condition during adolescence, a generally positive message emerges about these young people. CONCLUSIONS: Many young people with chronic illness appear relatively resilient in the face of the adjustment challenges presented by their illness. Interventions that allow a young person to explore meaning, build self-esteem, and acceptance through positive social connections are likely to improve adjustment outcomes in this group. PMID- 12625100 TI - From recreation to creative expression: the essential features of an adolescent inpatient psychosocial support programme. AB - INTRODUCTION: Adolescent Inpatient Units are now an established component of leading tertiary children's hospitals in the developed world. While different methods have been used to provide psychosocial support in these settings, there is little recent literature about these programmes. We aimed to identify programme goals and to explore the features that are thought to underpin an effective programme. MATERIAL AND METHODS: A qualitative paradigm was used to identify the essential features of the psychosocial support programme within the adolescent inpatient unit, Royal Children's Hospital, by drawing on the practice wisdom of experts involved in the development of the psychosocial support programme since its inception over a decade ago. In-depth interviews were conducted that were analysed thematically following recording and transcription. RESULTS: Seven out of nine identified professionals were interviewed. These staff identified that while the psychosocial support programme had focussed on different activities since its inception, the goals of the program had been consistent overtime. The essential underpinning features were reported to be support for the programme and communication between multidisciplinary health professionals. When this was effective, the overarching role of the programme was reported to be the promotion of identity formation in adolescents. CONCLUSIONS: This small-scale qualitative investigation reveals that identity formation has consistently been the focus of the psychosocial support programme in our adolescent unit, despite the different ways the programme has been run over time. Findings about the importance of support and communication are essential knowledge for those considering the implementation of similar programmes. PMID- 12625099 TI - Adolescent admissions to a tertiary paediatric hospital: a dynamic pattern. AB - INTRODUCTION: Adolescent inpatient facilities emerged in Australia in the 1980s to cater for an increasing number of young people with chronic illness and disability. Yet, there is minimal published data on the number of young people admitted to hospital within these units, their unit of admission, length of stay or reason for admission. A 12-year audit of adolescents admitted to a tertiary hospital with a dedicated adolescent unit and adolescent medicine programme in Victoria, Australia, was conducted to review the pattern of hospitalisation in young people in order to provide data to assist healthcare policy and planning agendas. MATERIALS AND METHODS: Admissions to the Royal Children's Hospital in Victoria, Australia, of adolescents aged 10 years and above were reviewed over a 12-year period from 1990 to 2001. We identified the annual number of adolescents admitted, the proportion of adolescents admitted to the Adolescent Inpatient Unit (ward) and annual admissions under the Adolescent Medicine Unit (department). RESULTS: Adolescents now constitute nearly 30% of all admissions at this children's hospital. Over this period, admissions to the Adolescent Inpatient Unit have nearly doubled and annual admissions under the Adolescent Medicine Unit rose from 38 to 288. The majority of adolescents were admitted under specialty medical and surgical units. CONCLUSIONS: The knowledge that nearly one in three admissions to this tertiary children's hospital is over 10 years old should help promote the development of planning and policy agendas that better balance both health and developmental priorities in this age group. PMID- 12625101 TI - Art and "the language of well-being" in adolescent health care. AB - Feeling effective as a young person depends on a capacity to draw upon one's own resources in the service of healthy living and development. In adolescent health care, there is the need to call upon the talents and creativity of young people, to introduce new and exciting experiences, and to facilitate involvement in their own care in order to nurture optimal growth and development on a physical and psychological level. While hospitalisation can represent a major crisis point in adolescence, the provision of a stimulating environment and the opportunity for creative activities offers an exciting, transformative and healing experience. Art allows adolescents to use alternative languages beyond illness, to engage in endeavours that are distanced from overt therapeutic intent, and to embrace attributes of self-esteem and resilience. Through the process and production of art, and the inclusion of music, poetry, film or theatre, young people can experience personal growth, acquire skills, develop socially and contribute to environmental change. In seeking to illustrate the value and importance of such approaches, this paper draws upon the experiences of a youth arts program attached to an adolescent ward. In a project called Art Injection, art students worked with adolescents to make sculptures from old hospital equipment, with startling results. More recently, the development of personal totem poles and an imaginative mosaic mural has powerfully engaged creativity and community in care. Group and individual art sessions, including the media arts project Creative Well, are offered on weekdays as part of the general hospital routine, enabling hospitalised young people to experience creativity as a daily part of their lives. PMID- 12625102 TI - Adolescent health education programmes: theoretical principles in design and delivery. AB - The patterns of health need in youth have changed over the last three decades with increasing rates of psychosocial problems compromising adolescent development and affecting the patterns of morbidity and mortality in adult years. This has increased the public health interest in health promotion, early detection and preventive health care for adolescents. Yet, young people report significant barriers to accessing the health care system, and health professionals report a self-perceived lack of competency and prior training in adolescent health, especially in these areas of youth health need. There is an imperative to provide quality professional development for existing health professionals and undergraduates in adolescent health care. This paper will discuss a model of professional knowledge in adolescent health and the evidence based principles in design and delivery of effective education programmes. It will also review the published evaluations of adolescent education programmes and outcomes that still require evaluation in the future. PMID- 12625103 TI - Clinical utility of selective intra-arterial calcium-stimulated hepatic venous sampling in regionalisation of insulinomas--the Singapore experience. AB - INTRODUCTION: To evaluate the efficacy and safety of selective intra-arterial calcium-stimulated hepatic venous sampling (ASVS) as a preoperative regionalisation modality for insulinomas. DESIGN AND METHODS: Four patients with biochemically-proven endogenous hyperinsulinism and negative spiral computed tomographic (CT) pancreas were subjected to ASVS between October 1999 to May 2001. Results obtained from ASVS were compared with localisation studies using either magnetic resonance imaging (MRI) or endoscopic ultrasonography (EUS); and these were confirmed surgically whenever possible. RESULTS: ASVS led to a definitive regionalisation in all 4 patients evaluated; the predicted location of the insulinoma matched the findings intraoperatively in all 3 patients who were operated upon. These were all proven to be insulinoma histologically. However, 1 patient showed a positive ASVS result in samples obtained from the left hepatic vein only. In the patient who was unable to undergo surgical resection due to other co-morbid factors, his ASVS findings were corroborated by localisation obtained from the MRI study. Conversely, EUS was found to give an incorrect localisation of insulinoma in 1 patient. Adverse effects were encountered in 3 patients (2 with mild hypotension and 1 with transient atrial fibrillation); however, premature termination of the procedure was not necessary in any of the patients. CONCLUSION: ASVS is accurate and reliable for regionalisation of insulinoma, especially in patients who do not have an obvious, unequivocal tumour using high quality current-generation MRI scans. PMID- 12625104 TI - Improving the uptake of human immunodeficiency virus (HIV) antenatal screening in a primary healthcare setting. AB - INTRODUCTION: It has been well established that proper treatment and management of human immunodeficiency virus (HIV)-infected mothers can decrease virus transmission to their fetuses. HIV screening of antenatal mothers attending government polyclinics was offered from 1998 at a subsidized cost of $5 and given on a voluntary basis. The polyclinics provide antenatal care to approximately 12% of pregnant mothers in Singapore who belong mainly to the lower socio-economic group. The uptake of the HIV screening by these mothers has been poor. MATERIALS AND METHODS: A one-month study was first undertaken in 1999 to determine the reasons for the poor uptake and to propose recommendations for improving the uptake. A second one-month study was undertaken in 2001 to evaluate the effect of the new strategies after implementation. FINDINGS: This first study showed that refusal for the test was mainly due to poor perception of risk among the mothers. The HIV screening test was incorporated into an antenatal screening package and consent was obtained for this package. This was implemented on an opt-out basis. The second study done after implementation showed that the uptake of the HIV screening improved from 45% to 99%. CONCLUSION: The incorporation of the antenatal HIV screening into an antenatal screening package, and implementing this on an opt-out basis, improves uptake to almost a universal level. PMID- 12625105 TI - Thyroid dysfunction in elderly patients. AB - INTRODUCTION: The aim of this study was to determine the prevalence of thyroid dysfunction in an elderly in-patient population in a restructured hospital. METHODS: This was a prospective observational study performed on consecutive patients admitted into a geriatric ward in a restructured hospital in Singapore over a period of 3 months. Thyroid function tests (free thyroxine and thyroid stimulating hormone) were performed on all patients during routine blood screening. For those with abnormal thyroid function tests, further investigations including thyroid autoantibodies and anterior pituitary hormone measurements were performed where indicated. Demographic data, the presence of sepsis, hypoalbuminaemia and patients' functional status as well as other thyroid-related data were captured. RESULTS: A total of 184 patients were screened and 62 (33.7%) patients were found to have abnormal thyroid function tests. The mean age was 83.8 years (SD 6.2). Twenty (32.3%) were males and 42 (67.7%) were females. Thirty-three (53.2%) patients had sepsis on admission, 29 (46.8%) were functionally dependent and 22 (35.5%) had hypoalbuminaemia. The prevalence of thyroid disorders were as follows: 1 (0.5%), 4 (2.2%), 9 (4.9%), 15 (8.2%) and 33 (17.9%) patients had hyperthyroidism, subclinical hypothyroidism, primary hypothyroidism, secondary hypothyroidism and sick euthyroid syndrome, respectively. Cross-tabulating sick euthyroid syndrome against functional dependence, hypoalbuminaemia and sepsis did not reveal any significant association (Fisher's exact test, P = 0.44, P = 0.42 and P = 0.61, respectively). CONCLUSION: There was a high prevalence of thyroid dysfunction in the elderly in patient population. We would advocate a lower threshold for screening elderly in patients for thyroid dysfunction. PMID- 12625106 TI - Immediate and mid-term safety and efficacy of single lead VDD pacemakers for patients with atrioventricular block and normal sinus node function--a single centre experience. AB - INTRODUCTION: Single lead atrial synchronous ventricular pacing (VDD) is increasingly being used in place of conventional dual chamber pacing (DDD) for patients with atrioventricular block and preserved sinus node function. Compared to the latter, VDD pacemakers provide similar haemodynamic benefits derived from atrial synchronous pacing, with the added benefit of an easier implant procedure. OBJECTIVE: To review the use, safety and efficacy of VDD pacing in a single tertiary referral centre. MATERIALS AND METHODS: A review of all patients with atrioventricular block and normal sinus node function implanted with a VDD pacemaker over a 2-year period at a local tertiary cardiac referral centre. Data on complications, atrial sensing performance and maintenance of atrioventricular synchrony during implant and at subsequent follow-up visits were obtained from a prospectively maintained registry and analysed. RESULTS: Forty-one patients (17 males, 24 females) with a mean age of 72 +/- 9 years received VDD pacing for various forms of high-grade atrioventricular block. The average implantation time was 46.8 +/- 17.1 minutes, and a pneumothorax in 1 patient was the only complication. Electrical measurements at implantation and subsequent follow-up visits revealed an initial rapid decrease in atrial signal amplitude (mean atrial P wave at implant 3.1 +/- 1.1 mV, predischarge 1.9 +/- 1.3 mV) which began to stabilise after 3 months, reaching a mean atrial P wave value of 1.3 +/- 0.3 mV at 24 months. The atrial sensing performance (percentage of atrial synchronous ventricular complexes) was 97% over a mean follow-up period of 9.9 months. Four patients (10%) developed paroxysmal atrial tachyarrhythmias. Sinus node dysfunction was not observed in any of our patients during the follow-up period. CONCLUSION: In patients with atrioventricular block and preserved sinus node function, single lead VDD pacing is safe and effective in maintaining a physiological atrial synchronous pacing mode. PMID- 12625107 TI - One-stage surgical correction of proximal hypospadias. AB - INTRODUCTION: Proximal hypospadias poses major problems in surgical correction owing to the complexity and severity of the abnormalities, leading to the use of staged repairs to correct this condition. However, with precise definition of the components of this deformity a single-stage operation can be developed and applied successfully for surgical correction of this condition. MATERIALS AND METHODS: Twenty-six patients with severe proximal hypospadias were subjected to a one-stage repair. Excision of proximal fibrotic dartos tissue and removal of dysplastic urethral plate tissue corrected chordee completely. Urethral reconstruction was then performed by tubularising a flap of dorso-lateral preputial skin which was then anastomosed to the proximal urethra. The glandular part of the urethra was reconstructed using the distal part of the flap as an onlay graft over the meatal groove. The suture lines were covered with a layer of dartos tissue and skin closure was completed by transposing dorsal skin to surface the ventral penile shaft. A urethral catheter was left in for 10 days. RESULTS: All patients have been followed up after surgery from 1 to 5 years with a median period of 2 years. There were no fistulas. Two patients had mild stenosis at the meatus which responded to dilatation. One patient developed a stricture at the proximal anastomosis which required secondary correction. All other patients achieved satisfactory correction, both in terms of voiding and in the cosmetic appearance of the genitalia. CONCLUSION: Single-stage repair of hypospadias can be successfully applied in the correction of severe proximal hypospadias. It requires meticulous dissection and careful design of reconstructive techniques. The end results are comparable to staged procedures and morbidity is significantly lower. PMID- 12625108 TI - Nipah encephalitis outbreak in Malaysia. AB - INTRODUCTION: Between September 1998 and June 1999, there was a severe outbreak of viral encephalitis among the pig farm workers in Malaysia. METHODS: This is a review of the published literature related to the outbreak with the focus on human diseases. RESULTS: The encephalitis was caused by a newly discovered paramyxovirus related to Hendra virus, later named Nipah virus. There were 265 patients with acute encephalitis. The disease is thought to spread from pig to man through close contact. The risk of human-to-human spread is thought to below. The disease affected mainly adult Chinese males, half of whom had affected family members. The disease presented mainly as acute encephalitis with a short incubation period of less than two weeks, with the main symptoms of fever, headache, and giddiness followed by coma. Distinctive clinical signs include segmental myoclonus, areflexia and hypotonia, hypertension, and tachycardia. Initial cerebrospinal fluid was abnormal in 75% of patients. Serology was helpful in confirming the diagnosis. Magnetic resonance imaging showed distinctive changes of multiple, discrete, and small high signal lesions, best seen with fluid-attenuated inversion recovery (FLAIR) sequences. Mortality was high at 40% and death was probably due to severe brainstem involvement. The main necropsy finding in acute encephalitis was that of disseminated microinfarction associated with vasculitis and direct neuronal involvement. Ribavirin was able to reduce the mortality by 36%. Relapse encephalitis was seen in 7.5% of those who recovered from acute encephalitis, and late-onset encephalitis in 3.4% of those with initial non-encephalitic or asymptomatic diseases. The mean interval between initial illness and the onset of the complication was 8.4 months. The relapse and late-onset encephalitis which manifested as focal encephalitis arose from recurrent infection. CONCLUSION: Nipah virus, a recently discovered paramyxovirus, causes a unique encephalitis with high mortality as well as relapse and late-onset encephalitis. The infection is mainly spread from pigs to man. PMID- 12625109 TI - Intermittent epoprostenol infusions in systemic lupus erythematosus associated pulmonary hypertension--a series of three cases. AB - INTRODUCTION: Pulmonary hypertension (PHT) associated with systemic lupus erythematosus (SLE) has a grave prognosis. Continuous prostacyclin infusion significantly reduces pulmonary arterial pressure (PAP) and may improve survival in patients with primary and secondary PHT. We report our experience with intermittent epoprostenol (EPO) infusion in SLE patients with PHT. CLINICAL CHARACTERISTICS: We reviewed patients with SLE associated PHT who were treated with intermittent EPO infusions in 1998. All 3 patients had severe PHT, with maximum systolic PAP (PASP) of 58, 96 and 67 mm Hg, respectively, when measured using Doppler echocardiography, and were in New York Heart Association functional class III. TREATMENT: All patients were given 6 infusions of EPO monthly via peripheral venous access without significant side effects. OUTCOME: The PASP of all patients remained stable during the therapy. The therapeutic response as measured by patient symptoms and PASP was variable. No patient had significant reduction of PASP during the 6-month treatment period. Two patients developed rebound elevation of PASP after cessation of treatment, while the PASP of the other patient remained stable for 10 months. CONCLUSION: Although intermittent EPO infusion did not substantially reduce PASP, it prevented further rise in PAP during the treatment period, suggesting that this regimen may benefit a subset of SLE patients with PHT. PMID- 12625110 TI - Unusual case of bowel infarction with invasive Aspergillus in an immunocompromised patient. AB - INTRODUCTION: We report a patient with bowel infarction due to invasive Aspergillus infection as a result of prolonged chemotherapy-induced neutropenia. CLINICAL PICTURE: The patient was given neoadjuvant chemotherapy with doxorubicin, docetaxel, and capecitabine for a breast tumour. She developed prolonged neutropenia, Escherichia coli and Candida krusei sepsis, acute arterial thrombosis of the left lower limb, and invasive aspergillus infection. TREATMENT: She underwent a subtotal colectomy, small intestine resection and an above-knee amputation. OUTCOME: The hospitalisation was complicated with methicillin resistant Staphylococcus aureus pneumonia and short gut syndrome. She subsequently underwent simple mastectomy with axillary clearance and received adjuvant chemotherapy and radiation without complication. CONCLUSION: Chemotherapy should be protocol-directed. In a non-trial situation, protocol must have a sound basis, proven safety, and efficacy. Aspergillus infection is uncommon in patients with solid tumours; prompt treatment must be started on high suspicion. PMID- 12625111 TI - Routine abdominal and pelvic ultrasound examinations: an audit comparing radiographers and radiologists. AB - An audit of 202 routine abdominal and pelvic ultrasound examinations was carried out to evaluate the clinical performance and interpretation of these scans by radiographers and compare them with radiologists, and to investigate the extended role of radiographers in performing these scans. Each scan was first performed by a radiographer and then repeated by the radiologist. The findings were subsequently compared and any discrepancy resolved by re-scanning the patient with or without the involvement of an independent radiologist, or by follow-up of the patient by other imaging studies. In 158 (78.2%) scans, there was complete agreement between the radiographer's and radiologist's findings. In 44 scans (21.8%), there was incomplete agreement--there were 108 abnormal findings in these scans with incomplete agreement/discrepancy in 53 abnormalities. Overall, the accuracy of radiographers was 92.0% and radiologists was 91.7%; however, the accuracy rates were 94.0% and 96.4%, respectively, when minor abnormal findings without significant influence on the patient's clinical outcome were excluded. PMID- 12625112 TI - The diagnosis and management of hypercalcaemia. AB - INTRODUCTION: Hypercalcaemia is a relatively common clinical problem with a wide spectrum of presentations. This review provides an overview of the diagnosis and management of hypercalcaemia, and recent developments in drug therapy. METHODS: A non-systematic review of the English-language literature on various aspects in the management of hypercalcaemia was conducted. RESULTS: Increasingly, more patients with asymptomatic or minimally symptomatic disease are being detected. Appropriate evaluation of hypercalcaemia requires a careful elucidation of the underlying causes, the commonest being primary hyperparathyroidism and malignancy. The decision to treat and extent of treatment depends on factors such as the degree of hypercalcaemia, severity of symptoms, and the underlying cause. Several options are readily available to lower serum calcium levels. In the setting of acute, symptomatic hypercalcaemia, general measures such as saline rehydration and loop diuretics, and more specific measures including bisphosphonates (e.g. clodronate, pamidronate, ibandronate and zoledronate), calcitonin, mithramycin, gallium, glucocorticoids and dialysis, could be utilised. Definitive correction of the underlying cause, such as surgery for primary hyperparathyroidism or anti-tumour therapy, is important in controlling chronic hypercalcaemia. Medical therapy (estrogen, alendronate and raloxifene) is available to control bone sequelae in asymptomatic, mild hyperparathyroidism where surgery is not indicated. CONCLUSION: The therapeutic approach to hypercalcaemia should be tailored to the clinical setting. Developments in the understanding of the pathophysiological processes and the increasing availability of more powerful therapeutic options would help in achieving good outcomes in the management of acute and chronic hypercalcaemia. PMID- 12625113 TI - [Molecular biology for physicians]. PMID- 12625114 TI - [The human genome and gene expression]. AB - Recent data estimate that the human genome contains only about twice as many genes as the c. elegans and drosophila genomes. However, this may not reflect the true difference in the complexity of the organisms, since splicing and other postranscriptional mechanisms may occur more frequently in humans. This review highlights the current knowledge of human gene structure and gene expression, with focus on mechanisms that play a role in human diseases. PMID- 12625115 TI - [Genetic counseling and genetic services]. AB - Advances in knowledge about the genetic basis of disease have lead to an increasing demand for and expectation of genetic counselling and genetic services. This presents a new challenge to the health care system, as well as a need for professional education and public dialogue. In this review the current provision of genetic counselling and services is described, especially with the focus on the process of counselling and the persons presenting for counselling. Genetic counselling is seen as an information process and the scenarios for decision-making are discussed. The future development of genetic services should be developed within an ethical framework and the importance of quality assessment is underlined. PMID- 12625116 TI - [Use of molecular biological methods in invasive prenatal genetic diagnosis]. AB - In the past, prenatal cytogenetic analysis was limited by answering times of one to three weeks and lack of exact diagnosis of some structural abnormalities. The number of prenatal analyses of monogenic diseases was small due to lack of knowledge of the gene and the mutation in question. The introduction of molecular biological techniques allows prenatal diagnosis of the most frequent trisomies within one to two days and exact diagnosis of almost all structural abnormalities. The Human Genome Project now allows prenatal diagnosis for most monogenic diseases, and the rapid improvement of the DNA-chip technology will increase the number of prenatal diagnoses even further. PMID- 12625117 TI - [Polymerase chain reaction--application of the technique]. AB - Our knowledge of genes and genetic diseases has increased tremendously since the polymerase chain reaction technique (PCR) was introduced as a technique in molecular biology. The technique has become a critical tool for basic research, molecular biology, and biotechnology. PCR has been the link between science and molecular biology as a diagnostic tool. High sensitivity within a short time and low labour cost have been the key elements for the success of PCR. Furthermore, combinations with other techniques in molecular biology have opened up for still new perspectives as e.g. fluorescence-based real-time reverse transcription PCR (RT-PCR). In addition to providing a snapshot of the state-of-the-art applications of PCR, a look is made into the new DNA-micro-array era. PMID- 12625118 TI - [DNA microarrays (DNA chips) used in molecular medical research]. AB - Microchip technology and mapping of the human genome has led to the invention of DNA-micro-array technology. In that technology thousands of genes are analysed in parallel on a small glass surface. The very large data material that is generated requires bioinformatic analysis in order to be transformed to understandable biological results. There are different forms of DNA-micro-arrays. Some are used for measuring gene expression, i.e. which genes are turned on and which genes are turned off. Others are used for sequencing, and finally some are used for measuring single nucleotide polymorphisms (SNPs). Some application possibilities of the arrays within the area of cell biology and classification of diseases are presented. Especially within the area of classification of cancer diseases the technology is promising, and a scientific breakthrough could occur in the area of classification of very aggressive cancers. PMID- 12625119 TI - [Apoptosis in chronic inflammatory bowel disease. The importance for pathogenesis and treatment]. AB - Inflammatory bowel disease (IBD) is characterized by the increased survival of lamina propria leucocytes and ulcerations and an increased epithelial leakiness that compromise the barrier function of the epithelial lining. Deregulated apoptosis seems to be a major cause of the impaired barrier function and of leucocyte survival. This review focuses on the regulation of epithelial and leucocyte apoptosis in the IBD colon. Furthermore, the implications for present and future therapeutic strategies are discussed. PMID- 12625120 TI - [Transgenic mice as disease models]. AB - Transgenic animal models have proven to be useful tools in understanding both basic biology and the events associated with disease. Recent technical advances in the area of genomic manipulation in combination with the availability of the human and murine genomic sequences now allow the precise tailoring of the mouse genome. In this review we describe a few systems in which transgenic animal models have been employed for the purpose of studying the etiology of human diseases. PMID- 12625121 TI - [Cloning techniques for mitochondrial diseases and prenatal diagnosis]. AB - Disorders caused by mutation in the mitochondrial DNA are uncommon. Due to the special pattern of inheritance and of the variability of penetrance the options to affected couples to have healthy children are few. So far traditional prenatal diagnosis is of limited benefit. The problems may be overcome by oocyte donation. However, if the couple wants their own biological offspring, no good method is available today. We discuss ooplasmic and nuclear transfer as possible future options for these couples to have healthy biological children. PMID- 12625122 TI - [Conformational diseases]. AB - Conformational diseases are diseases where cellular functions are compromised because of misfolded proteins. The conceptional framework of conformational diseases is found in the cellular protein quality control systems which in the normal and young cell eliminate misfolded proteins. Many inherited genetic defects result in the misfolding of proteins, which may lead to recessive disorders if the proteins in question are totally or partly eliminated or to dominant diseases if the proteins slip through the protein quality control and accumulate in the cell. These inherited diseases are all early onset. Misfolding may also occur in proteins with an intrinsic ability to aggregate and in oxidatively damaged proteins, which accumulate by ageing. If the protein quality control systems are not sufficiently efficient cell toxic protein complexes may accumulate. This pathogenesis is a major contributing factor in the development of late onset neurodegenerative disorders. PMID- 12625123 TI - [Monogenic hereditary diseases]. AB - Monogenic disorders are caused by the inheritance of single gene mutations; alternatively, a monogenic disorder arises as a consequence of a de novo mutation in either the paternal or maternal germ line. The exponential increase in our understanding of the human genome has resulted in the localisation and cloning of a vast number of disease genes which has enabled the precise characterization of the underlying molecular defect in many of these disorders. Single nucleotide substitutions and microdeletions are the major types of disease-related mutations, but more complex mutations have also been described. PMID- 12625124 TI - [Nephrectomy in metastatic renal cell carcinoma--an old myth in new clothes?]. PMID- 12625125 TI - [Therapy-requiring mental disorders among asylum seeking children]. PMID- 12625126 TI - Schistosomiasis and soil-transmitted helminth infections: forging control efforts. PMID- 12625127 TI - Royal Society of Tropical Medicine and Hygiene meeting at Manson House, London 17 January 2002. Cervical cancer in developing countries. AB - The public health importance of cervical cancer is now increasingly appreciated as a means to improve the general health of women in many developing countries. Developing countries account for 80% of the world burden, mostly due to the lack of effective control programmes. Infection with oncogenic types of human papillomaviruses (HPV) has been established as the central cause for cervical cancer. Thus, vaccination against HPV is a potentially useful strategy for prevention, but this may take several years to become a reality. Currently, early detection and treatment is the most effective approach to control cervical cancer. Cervical cancer may be controlled through improving awareness and accessibility to diagnostic and treatment services. Cytology-based screening is beyond the capacity of health services in many developing countries, hence, alternative methods to cytology are being investigated. Visual inspection of the cervix after application of 3-5% acetic acid (VIA) seems to be a promising screening test, with a similar sensitivity to that of cytology, but lower specificity. Currently, it is being evaluated for its cost-effectiveness in reducing cervical cancer incidence and mortality in randomized trials. Information from the ongoing studies will be valuable for evolving cervical cancer control policies and programmes in low-resource settings. PMID- 12625129 TI - Zooprophylaxis, artefact or reality? A paired-cohort study of the effect of passive zooprophylaxis on malaria in The Gambia. AB - The World Health Organization has recommended the use of cattle for zooprophylaxis as a protective measure against malaria since 1982. However, concern has been raised about this practice, since some studies have shown that the presence of cattle may instead increase malaria prevalence. This study was designed to investigate the effect of passive zooprophylaxis on malaria in an area of moderate seasonal transmission in The Gambia, West Africa. The study was based on a paired-cohort of 204 children aged < 7 years, sleeping < 20 m or > 50 m from cattle, and surveys were done from 14 October to 2 December 1997. Entomological investigations showed that the presence of cattle did not alter the risk of malaria transmission in nearby houses. There was also no significant difference in the prevalence of Plasmodium falciparum between the 2 groups. Although the presence of cattle appeared to be protective against high parasitaemia, cattle were also associated with greater wealth of the children's families. Conditional logistic regression analysis showed that the decreased risk of high parasitaemia in the group with cattle present was an artefact associated with the higher general wealth of the cattle owners. We concluded that zooprophylaxis is not an effective intervention method against malaria in settings similar to The Gambia. PMID- 12625128 TI - Risk factors for malaria in pregnancy in an urban and peri-urban population in western Kenya. AB - To assess risk factors for malaria in pregnancy in Kisumu, western Kenya, we studied healthy women with an uncomplicated pregnancy of > or = 32 weeks attending the antenatal clinic in the Provincial Hospital. Between June 1996 and March 1999, malaria and human immunodeficiency virus (HIV) infection were examined in 5093 pregnant women: 20.1% of the women were parasitaemic and 24.9% were HIV-seropositive. 2502 women delivered in the hospital and a smear was obtained: the prevalence of placental malaria, maternal peripheral parasitaemia, and HIV infection was respectively 19.0%, 15.2% and 24.5%. HIV infection (risk ratio [RR] 1.58, 95% confidence interval [95% CI] 1.32-1.89), young age (< 21 years: RR 1.51, 95% CI 1.19-1.91), being a primigravidae (RR 1.41, 95% CI 1.05 1.88), a peri-urban residence (RR 1.50, 95% CI 1.21-1.88), and Luo ethnicity (RR 1.74, 95% CI 1.35-2.24) were risk factors for malaria at delivery. Use of sulfadoxine-pyrimethamine (SP), reported by 2.1% of the women, was a protective factor (RR 0.44, 95% CI 0.18-1.06). Results were similar in the third trimester. In this urban/peri-urban setting, preventing HIV infection, delaying the first pregnancy until after adolescence, and applying an effective antimalarial strategy such as intermittent therapy with SP will reduce the prevalence of malaria in pregnancy. PMID- 12625131 TI - Effect of water supply and sanitation on the prevalence and intensity of Ascaris lumbricoides among pre-school-age children in Ajebandele and Ifewara, Osun State, Nigeria. AB - The relationship between Ascaris lumbricoides infection and provisions for safe water supply and excreta disposal was investigated in Ajebandele and Ifewara, 2 peri-urban communities near Ile-Ife, Osun State, Nigeria, from September 1998 to December 1999. Although there were more facilities for safe water supply (pipe borne and shallow wells) and sanitation (flush toilets and pit latrines) in Ajebandele than in Ifewara, they were not evenly distributed and only a few families, mainly in Ajebandele, were well provided for. Our results indicated that the prevalence of A. lumbricoides was significantly higher in Ifewara (21.2%) than in Ajebandele (13.1%), and within villages was much lower in houses with tap water and flush toilets. However, the mean intensity of infection was unexpectedly higher in Ajebandele (166.5 +/- 692.0) than in Ifewara (153.1 +/- 485.8) suggesting that the present level of investment in Ajebandele is probably not sufficient to reduce morbidity. These findings confirm suggestions that there is a threshold level of investment below which interventions on water supply and sanitation may not significantly improve human health. Sustainable ascariasis control in the study communities will require significant improvements to existing water supplies and sanitation facilities and any future interventions should incorporate a programme of health education to ensure that the facilities provided are properly used and hygienically maintained. PMID- 12625130 TI - Mortality in a seven-and-a-half-year follow-up of a trial of insecticide-treated mosquito nets in Ghana. AB - A 17% efficacy in preventing all-cause mortality in children aged 6-59 months was previously reported from a cluster-randomized controlled trial of insecticide treated mosquito nets (ITNs) carried out in the Kassena-Nankana District of northern Ghana from July 1993-June 1995. A follow-up until the end of 2000 found no indication in any age group of increased mortality in the ITN group after the end of the randomized intervention. These results should further encourage the use of ITNs as a malaria control tool in areas of high endemicity of Plasmodium falciparum. PMID- 12625132 TI - The intensity of transmission of hepatitis A and heterogeneities in socio environmental risk factors in Rio de Janeiro, Brazil. AB - The objective of this work was to assess the intensity of transmission of hepatitis A in Rio de Janeiro, Brazil. We also used the estimation of the parameters of a deterministic model to study the effects of risk factors. Age specific seroprevalence of antibodies against hepatitis A virus (HAV) was obtained from a survey screening in a city of the metropolitan area of Rio de Janeiro, in 1997. From the seroprevalence data, we estimated the age-dependent force of infection (lambda) and the average age of first infection (A), using a deterministic model. To evaluate the influence of the environmental risk factors, we estimated the same parameters stratifying the sample for the selected socio environmental risk factors: the number of years of schooling of the female responsible for the house, crowding within the bedroom, number of water taps and fittings, and the presence of sewage in front of the house. For the whole sample, the maximum force of infection estimated was 0.12/year and the average age of infection was 10.1 years. This last parameter decreased as the number of persons per bedroom increased, and also when the number of water taps and the number of years of schooling of the woman responsible for the house decreased. The proposed environmental interventions may lead to a decrease in the intensity of transmission of HAV and an increase in the average age of first infection in the next few years. This may have public health implications, since hepatitis A is more severe in adults. In this context, specific vaccination programmes may be necessary, as in developed countries. PMID- 12625133 TI - Vector competence of Brazilian Aedes aegypti and Ae. albopictus for a Brazilian yellow fever virus isolate. AB - Because the potential urban yellow fever (YF) mosquito vectors Aedes aegypti and Ae. albopictus are at historical highs in Brazil, both in terms of density and geographical range, we assessed the risk of an urban YF epidemic in Brazil. We evaluated and confirmed in a laboratory setting the vector competence of Brazilian Ae. aegypti for a currently circulating strain of YF virus, and investigated the potential for Brazilian Ae. albopictus to transmit YF. PMID- 12625134 TI - Care-seeking behavioural patterns, awareness and diagnostic processes in patients with smear- and culture-positive pulmonary tuberculosis in Lagos, Nigeria. AB - This study was carried out in 2 chest referral clinics in Lagos, Nigeria, between February 2000 and May 2001 to assess the effects of knowledge, attitude, and practice of 168 newly diagnosed tuberculosis (TB) patients on their care-seeking behaviour. At the onset of symptoms patients sought treatment from one or more of the local private orthodox and traditional health providers, and patent medicine dealers before presenting at a chest clinic. There was a correlation between the level of knowledge and awareness of TB with time of presentation at the chest clinic. Of the 32 patients who presented and were diagnosed at the chest clinics within 4 weeks of onset of symptoms, 50% had knowledge of the aetiological agent of the disease and 60% had some idea of the mode of transmission. Of the 105 patients who presented and were diagnosed 12 weeks after the onset of symptoms, 97% had no knowledge of the aetiological agent and 95% had no idea of the mode of transmission. Overall there was a low level of knowledge and awareness of the disease as well as an apparently high level of improper health care-seeking behaviour amongst the patients studied which probably contributed significantly to the delay in early and accurate diagnosis of most of the cases. These findings indicate an urgent need to educate communities and care providers on the cause and mode of transmission of TB, and the need to attend designated health facilities for early diagnosis and proper treatment. PMID- 12625135 TI - Sexually transmitted infections among prison inmates in a rural district of Malawi. AB - As part of a comprehensive human immunodeficiency virus (HIV) prevention strategy targeting high-risk groups, sexually transmitted infection (STI) clinics are offered to all prisoners in Thyolo district, southern Malawi. Prison inmates are not, however, allowed access to condoms as it is felt that such an intervention might encourage homosexuality which is illegal in Malawi. A study was conducted between January 2000 and December 2001 in order to determine the prevalence, incidence, and patterns of STIs among male inmates of 2 prisons in this rural district. A total of 4229 inmates were entered into the study during a 2-year period. Of these, 178 (4.2%) were diagnosed with an STI. This included 83 (46%) inmates with urethral discharge, 60 (34%) with genital ulcer disease (GUD), and 35 (20%) inmates with epididymo-orchitis. Fifty (28%) STIs were considered incident cases acquired within the prisons (incidence risk 12 cases/1000 inmates/year). GUD was the most common STI in this group comprising 52% of all STI. This study shows that a considerable proportion of STIs among inmates are acquired within prison. In a setting of same-sex inmates, this suggests inter prisoner same-sex sexual activity. The findings have implications for HIV transmission and might help in developing more rational policies on STI control and condom access within Malawi prisons. PMID- 12625136 TI - Anopheline vectors and malaria transmission in eastern Afghanistan. AB - Anopheline vectors and malaria transmission were studied in 2 river-irrigated, rice-growing districts of eastern Afghanistan from May 1995 to December 1996. Clinical malaria was monitored in 12 rural villages (population 14,538) by passive case detection at local clinics. Adult mosquitoes were collected by space spraying of living quarters and stables and by cattle bait catches. Mosquito head thoraces (17,255 specimens) were tested for Plasmodium falciparum and P. vivax circumsporozoite protein (CSP) using enzyme-linked immunosorbent assay. The recorded incidence of P. vivax and P. falciparum was 199 and 41 episodes per 1000 person years, respectively. Twelve species of anopheline were recorded; Anopheles stephensi comprised 82% and A. culicifacies 5%. Eight species tested positive for CSP: A. stephensi, A. culicifacies, A. fluviatilus, A. annularis, A. pulcherrimus, A. maculatus, A. splendidus and A. superpictus. Among infected mosquitoes 46% were positive for P. falciparum, 45% for P. vivax VK-247, and 9% for P. vivax PV-210. Estimates of the feeding rates of infective vectors on humans indicated that A. stephensi would contribute 76% of infective bites, A. fluviatilis and A. pulcherrimus 7% each, and A. culicifacies and A. superpictus 3% each. The overall infective vector feeding rate correlated with the P. vivax incidence rate in the human population. The conventional view of A. culicifacies being the main rural vector and A. stephensi important only in urban settings needs to be reconsidered in western outreaches of the Indo-Pakistan subcontinent. PMID- 12625137 TI - Visceral leishmaniasis in Tunisia: result of the isoenzymatic characterization of 65 Leishmania infantum strains. AB - The isoenzymatic characterization of 65 Tunisian strains of Leishmania, isolated from human cases of visceral leishmaniasis between June 1998 and August 2001, revealed the existence of 3 zymodemes of the L. infantum complex: MON-1 the most common (93.8%), followed by MON-24 (3.1%) and MON-80 (3.1%). 72% of the strains were obtained from children under the age of 5 years. The majority of the studied strains originated from 8 provinces of northern Tunisia, particularly the province of Zaghouan. PMID- 12625138 TI - American visceral leishmaniasis: wild animal hosts. AB - In Colombia it has been suggested that the rodent Proechimys canicollis may be a reservoir-host of Leishmania (L.) chagasi, the cause of American visceral leishmaniasis, based on polymerase chain reaction/hybridization tests. We have detected no infection in laboratory-bred specimens of another species, P. guyannensis, after their inoculation with promastigotes or amastigotes of this parasite from Amazonian Brazil. PMID- 12625139 TI - Deforestation and the spatio-temporal distribution of savannah and forest members of the Simulium damnosum complex in southern Ghana and south-western Togo. AB - Spatio-temporal data on cytotaxonomic identifications of larvae of different members of the Simulium damnosum complex collected from rivers in southern Ghana and south-western Togo from 1975 until 1997 were analysed. When the data were combined, the percentages of savannah blackflies (S. damnosum sensu stricto and S. sirbanum) in the samples were shown to have been progressively increasing since 1975. The increases were statistically significant (P < 0.001), but the rates of increase were not linear. Further analyses were conducted according to the collection seasons and locations of the samples, to account for possible biases such as savannah flies occurring further south in the dry season or a preponderance of later samples from northern rivers having more savannah flies. These analyses showed that the increasing trend was statistically significant (P < 0.0001) only during the periods April to June and October to December. The presence of adult savannah flies carrying infective larvae (L3) indistinguishable from those of Onchocerca volvulus in the study zone was confirmed by examinations of captured flies. The percentages of savannah flies amongst the human-biting populations and the percentages with L3s in the head were higher during dry seasons than wet seasons and the savannah species were found furthest south (5 degrees 25'N) in the dry season. Comparisons of satellite images taken in 1973 and 1990 over a study area in south-western Ghana encompassing stretches of the Tano and Bia rivers demonstrated that there have been substantial increases in urban and savannah areas, at the expense of forest. This was so not only for the whole images but also for subsamples of the images taken at 1, 2, 4, 8 and 16 km distant from sites alongside the River Tano. At every distance from the river, the percentages of pixels classified as urban or savannah have increased in 1990 compared with 1973, while those classified as degraded or dense forest have decreased. The possibility that the proportionate increases in savannah forms of the vectors of onchocerciasis, and hence in the likelihood of the transmission of savannah strains of the disease in formerly forested areas, were related to the decreases in forest cover is discussed. PMID- 12625140 TI - Detection of Giardia duodenalis antigen in coprolites using a commercially available enzyme-linked immunosorbent assay. AB - The objective of this experiment was to assess the utility of a commercially available enzyme-linked immunosorbent assay (ELISA) kit for diagnosis of giardiasis in archaeological human remains. The kit, a monoclonal antibody assay, is used to detect the presence of Giardia-specific antigen 65 (GSA65) in human faeces. We utilized the assay in ancient faecal material. The material included desiccated faeces found in mummies or in archaeological sites, and sediments from latrines. A total of 83 specimens, previously examined microscopically for parasites, were examined. The ELISA detected 3 positive samples, dated to about 1200 AD, 1600 AD and 1700 AD. The ELISA was superior to direct observation. It was possible to identify G. duodenalis cysts by direct microscopy in only one of these samples. The results did not show cross-reactivity between this protozoan and helminths. The use of ELISA to detect G. duodenalis coproantigen could help the diagnosis of giardiasis in ancient human remains. PMID- 12625141 TI - Haemozoin as a marker of placental parasitization. AB - Both Plasmodium vivax and P. falciparum malaria can cause the delivery of low birthweight babies. In this report, we have quantitated haemozoin levels in placentas from women living on the Thai-Burmese border in a region of low transmission for both P. falciparum and P. vivax malaria from June 1995 to January 2000. P. falciparum malaria infections during pregnancy lead to the accumulation of haemozoin (malaria pigment) in the placenta, especially in infections near term and in primigravid pregnancies. Haemozoin concentration was not associated with adverse birth outcomes. Women with P. vivax infections during pregnancy do not have measurable levels of placental haemozoin suggesting that P. vivax-infected erythrocytes do not accumulate in the placenta as much as P. falciparum-infected ones. PMID- 12625142 TI - Case report: severe acute symptomatic hyponatraemia in falciparum malaria. AB - Hyponatraemia is a common finding in malaria, but rarely appears to be of clinical significance. We describe a case of acute, profound, hyponatraemia with confusion and convulsions developing in the context of seemingly uncomplicated Plasmodium falciparum malaria. We draw attention to this rarely reported and poorly documented life-threatening complication and review the limited literature on the subject. PMID- 12625144 TI - Case report: intraocular localization of Mansonella perstans in a patient from south Chad. AB - A case of Mansonella perstans infection with visual impairment and a retinal lesion is described. After a course of diethylcarbamazine and a further course of mebendazole the symptoms improved and the microfilariae blood concentration decreased. The symptoms and response to antifilarial treatment strongly suggested the intraocular localization of an M. perstans worm. PMID- 12625143 TI - Filaria dance sign and subclinical hydrocoele in two east African communities with bancroftian filariasis. AB - During population-wide cross-sectional surveys for Wuchereria bancrofti microfilaremia, circulating antigenaemia, and clinical disease in a high and a low endemicity community in East Africa in 1998, a portable ultrasound scanner was used simultaneously to examine the scrotal tissue of the male populations (n = 422 and 328, respectively) for signs of adult worms. The overall microfilaria (mf) and circulating filarial antigen (CFA) prevalences in the scanned males were 30.8% and 53.6% in the high and 4.3% and 19.8% in the low endemicity community, respectively. During ultrasound examination, the filaria dance sign (FDS)- indicating the presence of live adult W. bancrofti worms--was observed in 16.1% and 6.7% of the males in these communities, respectively. This examination also revealed that subclinical hydrocoele (fluid accumulation in the scrotal sac, not detected during physical examination for clinical hydrocoele) was very common, affecting 25.3% and 15.5% of the examined males in the high and low endemicity community, respectively. Both of these ultrasonographic signs started to appear around the age of puberty and were most common in adults. In the high endemicity community, the prevalence and mean intensity of mf and CFA were considerably higher in FDS-positive than in FDS-negative adult males, whereas no obvious difference in these parameters was noted between adult males with and without subclinical or the combination of clinical and subclinical hydrocoele. Associations were less clear in the low endemicity community, probably because of the low number of infected individuals. The application of ultrasonography as a tool in bancroftian filariasis epidemiological field studies thus indicated that scrotal pathology may be much more common in endemic areas than hitherto reported. PMID- 12625146 TI - Reassessment of the resistance of Plasmodium falciparum to chloroquine in Gabon: implications for the validity of tests in vitro vs. in vivo. AB - Increasing resistance of Plasmodium falciparum to antimalarial drugs presents a major risk factor for people living in endemic areas of tropical Africa. In Lambarene, Gabon, regular surveillance of chloroquine sensitivity of P. falciparum in vitro has been carried out since 1992 using the WHO standard microtest. Results indicated that from 1994 onwards chloroquine resistance in vitro decreased significantly and that by 2000, about 70% of parasite isolates seemed to be sensitive to chloroquine in vitro. In 2001, we conducted a clinical study to reassess the efficacy of chloroquine in vivo for the treatment of uncomplicated P. falciparum malaria. Twenty-six patients aged 4-15 years were included in this study. Most unexpectedly, the study demonstrated high-grade resistance to chloroquine in vivo (failure rate on day 28 of 100%). As a consequence, tests of parasite susceptibility to chloroquine in vitro were repeated using the same protocol except for the replacement of previously used commercially available predosed WHO culture plates by independently dosed plates. All tested P. falciparum isolates were highly resistant to chloroquine, correlating well with our clinical findings. We concluded that high level resistance of P. falciparum to chloroquine persists in the study area. Neglect or absence of quality controls of essential test material can lead to invalid study results and wrong conclusions and should always be suspected in the case of major fluctuations in the sensitivity patterns of an antimalarial drug in vitro. In addition, our results highlight the supreme value of tests in vivo in providing reliable estimates of the efficacy of an antimalarial in a specific area. PMID- 12625145 TI - A randomized, double-blind study on the efficacy and safety of a practical three day regimen with artesunate and mefloquine for the treatment of uncomplicated Plasmodium falciparum malaria in Africa. AB - A randomized, double-blind, parallel-group study in 104 hospitalized patients with acute, uncomplicated Plasmodium falciparum malaria was performed in West and Central Africa from March to July 2001. Patients were randomized to receive simultaneous dosing (artesunate 200 mg/d plus mefloquine 250 mg/d from the first to the third day [investigational group]) or sequential dosing (artesunate 200 mg/d for 3 d plus mefloquine 250 mg on the second and 500 mg on the third day [reference group]). Patients were followed-up for 28 d, and clinical and parasitological outcomes were assessed. The 14-d cure rate was 100% in the investigational group and 98% in the reference group with no recrudescence until day 28. Mean times to fever and parasite clearance were similar between the 2 groups (32 h vs. 26 h and 45 h vs. 48 h) and tolerability was good in both groups. The number of patients with vomiting was statistically significantly lower in the investigational group compared to the reference group (3.8% vs. 19.2%, P = 0.014). A 3-d once-daily co-administration of artesunate and mefloquine starting on day one offers a practical dosing regimen, which is highly effective and well tolerated in patients with uncomplicated P. falciparum malaria. PMID- 12625147 TI - High Plasmodium falciparum resistance to chloroquine and sulfadoxine pyrimethamine in Harper, Liberia: results in vivo and analysis of point mutations. AB - In Liberia, little information is available on the efficacy of antimalarials against Plasmodium falciparum malaria. We measured parasitological resistance to chloroquine and sulfadoxine-pyrimethamine (SP) in Harper, south-west Liberia in a 28-d study in vivo. A total of 50 patients completed follow-up in the chloroquine group, and 66 in the SP group. The chloroquine failure rate was 74.0% (95% confidence interval [95% CI] 59.7-85.4%) after 14 d of follow-up and 84.0% (95% CI 70.9-92.8%) after 28 d (no polymerase chain reaction [PCR] analysis was performed to detect reinfections in this group). In the SP group, the failure rate was 48.5% (95% CI 36.2-61.0%) after 14 d and 69.7% (95% CI 57.1-80.4%) after 28 d, readjusted to 51.5% (95% CI 38.9-64.0%) after taking into account reinfections detected by PCR. Genomic analysis of parasite isolates was also performed to look for point mutations associated with resistance. Genotyping of parasite isolates revealed that all carried chloroquine-resistant K-76T mutations at gene pfcrt, whereas the triple mutation (S108N, N511, C59R) at dhfr and the A437G mutation at dhps, both associated with resistance to SP, were present in 84% and 79% of pretreatment isolates respectively. These results seriously question the continued use of chloroquine and SP in Harper and highlight the urgency of making alternative antimalarial therapies available. Our study confirms that resistance to chloroquine may be high in Liberia and yields hitherto missing information on SP. PMID- 12625148 TI - Efficacy of amodiaquine for uncomplicated Plasmodium falciparum malaria in Harper, Liberia. AB - In the face of spreading chloroquine and sulfadoxine-pyrimethamine (SP) resistance, amodiaquine remains a cheap and efficacious alternative for treating uncomplicated Plasmodium falciparum malaria in many settings. In Harper, south eastern Liberia, a previous study we conducted showed very high levels of resistance to both chloroquine and SP. In 2001, in an effort to look for possible alternatives, we measured in the same setting the efficacy of amodiaquine in a 28 d study in vivo, with results corrected by polymerase chain reaction genotyping to distinguish recrudescences from reinfections. In total, 107 children were included in the study and received a 3-d supervised course of 25 mg/kg amodiaquine. Of these, 81 were analysable at day 28. The overall failure rate was 19.8% (95% CI 11.7-30.1%) considering both parasitological and clinical outcomes. These results provide hitherto missing data on amodiaquine in Liberia, and confirm that the drug may still be efficacious in settings where chloroquine and SP are failing. We recommend the introduction of amodiaquine in association with artesunate as a first-line antimalarial in Harper. PMID- 12625149 TI - The effect of 10 alpha-trifluoromethylhydroartemisinin on Plasmodium berghei infection and its toxicity in experimental animals. AB - The antimalarial activity of 10 alpha-trifluoromethylhydroartemisinin (TFMHA) was compared to that of dihydroartemisinin (DHA) in the Plamodium berghei mouse model. Treatment with TFMHA in mice infected with a P. berghei chloroquine sensitive strain at 25 mg/kg for 3, 5, and 7 d, or DHA at the same dose for 7 d showed the parasite was eliminated from the host within 2.6 d. The radical cure and survival rates of these mice up to 60 d after infection were 90-100%. In mice infected with the P. berghei chloroquine-resistant strain, TFMHA used at 25 mg/kg/day for 3, 5, or 7 d reduced parasitaemia within 2 d. The radical cure and survival rates of these animals up to 60 d after infection were 30, 60, and 90% for the 3 treatment durations respectively. In contrast, DHA only had an inhibitory effect on the growth of the parasite within the first few days of treatment and could not eliminate the parasite even after 7 d of treatment. There was a 100% relapse and all mice died within 28 d after infection. The acute toxicity of TFMHA as determined by the median lethal dose (LD50) in mice treated orally was 820 mg/kg. In rabbits, TFMHA given orally at 20 mg/kg once daily for 28 successive days had no effect on the bodyweight, haematological, biochemical, histopathological and electrocardiogram parameters. The results showed that TFMHA is an effective antimalarial drug with a low level of toxicity. PMID- 12625150 TI - Comparison of tafenoquine (WR238605) and primaquine in the post-exposure (terminal) prophylaxis of vivax malaria in Australian Defence Force personnel. AB - On return from duty in North Solomons Province (including Bougainville Island), Papua New Guinea, 586 Australian Defence Force personnel received either primaquine (14-d) or tafenoquine (3-d) post-exposure malaria prophylaxis. Within 12 months, 6 of the 214 volunteers receiving primaquine and 7 of 378 receiving tafenoquine had developed vivax malaria. Overall, volunteers preferred the shorter course of tafenoquine. PMID- 12625152 TI - Height as a proxy for weight in determining azithromycin treatment for paediatric trachoma. AB - Azithromycin (Zithromax, Pfizer Inc., New York, NY, USA) is effective in the control of blinding trachoma. Community-based azithromycin treatment is recommended by the World Health Organization as part of a multipronged strategy aimed at the global elimination of binding trachoma by the year 2020. Paediatric trachoma is treated with azithromycin according to weight at a target dosage of 20 mg/kg. However, conventional weight-based treatment may be problematic in the field due to the logistical difficulties associated with weight scales. We assessed the accuracy of using height as a proxy for weight to determine azithromycin treatment in 4 countries--Viet Nam, Tanzania, Ghana, and Mali--where mass treatment programmes are underway. Population-based data collected from 1988 to 2000 were analysed using least squares regression. Height treatment schedules were developed for each data set. The accuracy of each schedule was evaluated according to the percentage of children receiving treatment within a dosage range of 20-30 mg/kg, a conservative estimate of the safe and effective treatment range for paediatric trachoma. Using height to determine dose, 89-95% of children would receive a dosage of 20-30 mg/kg. In these populations, height-based treatment is a reliable alternative to conventional weight-based treatment. Methods for developing height schedules presented in this analysis could be applied to other regions and therapeutics. PMID- 12625153 TI - Are health personnel the best choice for directly observed treatment in southern Thailand? A comparison of treatment outcomes among different types of observers. AB - A prospective study was conducted in 24 districts in southern Thailand in 1999 with directly observed treatment, short-course strategy (DOTS) implemented to determine treatment outcomes in relation to the practical observer among 455 enrolled patients with tuberculosis. Health personnel (HP), community members (CM), family members (FM) and self-administration (SA) were initially assigned to be DOT observers in 43%, 5%, 44% and 8% of 411 analysed patients, respectively. In practice, 56% of the 379 patients with assigned observers changed their observers. The practical observer was the assigned observer among 17% of patients assigned to HP, 57% to CM, 75% to FM, and 34% to SA, respectively. There were no significant differences in treatment success between different types of main observers. Adjusted odds ratios (95% confidence interval) of treatment non success were 1.1 (0.3-4.7), 0.7 (0.2-3.3), and 0.5 (0.2-1.1) for HP, CM, and FM, over SA groups, respectively. HP may not be the best choice in our setting due to poor sustainability and the availability of another promising choice (CM). PMID- 12625151 TI - Evaluation of the efficacy of pyrantel-oxantel for the treatment of soil transmitted nematode infections. AB - A randomized controlled trial comparing the efficacy of pyrantel-oxantel (10 mg/kg) with mebendazole (500 mg) was performed on 1329 schoolchildren aged 6-9 years on Pemba Island in September-October 2000 to evaluate alternative single dose drugs for regular treatment of intestinal nematode infections. Both mebendazole and pyrantel-oxantel were very effective in eliminating Ascaris lumbricoides infection, inducing cure rates of more than 96% and reducing the mean egg counts by more than 95%. Both drugs had a moderate efficacy against Trichuris trichiura infection, but pyrantel-oxantel had a higher cure rate (31.5% vs. 23.3%, P < 0.01), though the reductions in egg counts did not differ significantly and were more than 80%. Pyrantel-oxantel and mebendazole had a similar, poor efficacy in curing hookworm infections and had a moderate effect in reducing the egg counts by 67% and 68%, respectively. Pyrantel-oxantel (10 mg/kg) offers a valuable alternative to mebendazole as a single-dose treatment for the control of intestinal nematode infections in children in endemic areas of sub Saharan Africa, due to its comparable efficacy, its low cost and its suitability for use in young children. PMID- 12625154 TI - ABO blood groups, grass pea preparation, and neurolathyrism in Ethiopia. AB - An exploratory study was conducted in the rural Estie district of Ethiopia in 1997 to identify the role of ABO blood group, rhesus factor, and type of grass pea (Lathyrus sativus) diet in the susceptibility to neurolathyrism. Five-hundred study subjects (250 cases and 250 controls) were examined and interviewed, and had their ABO and rhesus blood groups determined. The majority (86%) of the cases were males. Blood group O was the most common in the patients and controls followed by groups A, B, and AB. The vast majority of the study subjects were rhesus-positive. The gravy (Shiro) grass pea preparation was consumed by 91.6% of the study population, boiled (Nifiro) by 86%, and roasted (Kollo) by 56.4%. Almost half (48%) of the cases had consumed grass pea for > 4 months compared to 8% of controls (P < 0.001). There was a significant association between the risk for neurolathyrism and the consumption of boiled (adjusted odds ratio [AOR] = 98.4) and roasted (AOR = 55.62) forms of grass pea. There was no risk of paralysis associated with consumption of the gravy form of grass pea (AOR = 0.40, 95% confidence interval 0.1-2.0). Blood group O remained significantly associated with the disease after adjusting for age, type of grass pea preparation consumed, and duration of consumption (AOR = 2.90). PMID- 12625155 TI - Malaria transmission, antimalarial-drug use, and resistance. PMID- 12625156 TI - [The use of disinfectants in veterinary practice]. AB - The use of chemical disinfectants within the veterinary practice is only permitted when the disinfectants are legally registered. A distinction has to be made between disinfectants to be used on the skin of men or animals and disinfectants to be used on surfaces like floors, walls, cages, stables and for veterinary instruments. For the first group, to be considered as (veterinary) medicines, the Act on (Veterinary) Medicines applies. For the second group, to be considered as veterinary biocides, the Pesticide Act applies. A small survey carried out by the Keuringsdienst van Waren shows that in veterinary practice disinfectants are applied in an inappropriate way. A lack of knowledge and the lack of hygienic protocols could be reasons for these misuses. In this article the Keuringsdienst van Waren gives information about the legal aspects of disinfectants that fall within the scope of the Pesticide Act. PMID- 12625157 TI - [Gastrointestinal diseases in the cat: are cats different from dogs?]. PMID- 12625158 TI - [Adverse effects of estradiol-17beta and alternatives for prostaglandins]. PMID- 12625159 TI - [Ammonium hydroxide instead of ammonium chloride]. PMID- 12625160 TI - [Research prostate carcinoma in the dog at the University Clinic for Companion Animals in Utrecht, the Netherlands]. PMID- 12625161 TI - [Senecio jacobaea]. PMID- 12625162 TI - [BSE: an additional explanation]. PMID- 12625163 TI - [Question marks about the use of enrofloxacin in sport horses]. PMID- 12625164 TI - Reflections on making decisions. PMID- 12625165 TI - Meeting unmet needs in patients with sepsis: the role of drotrecogin alfa (activated). PMID- 12625166 TI - Synergy model and the role of clinical nurse specialist in a multihospital system. PMID- 12625167 TI - Dying by irreconcilable numbers. PMID- 12625168 TI - Subjective perceptions and physiological variables during weaning from mechanical ventilation. AB - BACKGROUND: As costs related to mechanical ventilation increase, clear indicators of patients' readiness to be weaned are needed. Research has not yet yielded a consensus on physiological variables that are consistent correlates of weaning outcomes. Subjective perceptions rarely have been examined for their contribution to successful weaning. OBJECTIVE: To explore the subjective perceptions of dyspnea, fatigue, and self-efficacy and selected physiological variables in patients being weaned from mechanical ventilation. METHODS: Data were collected prospectively on 68 patients being weaned from mechanical ventilation. Subjective perceptions were measured by using 3 visual analog scales; physiological variables were measured by using the Burns Weaning Assessment Program and a patient profile. Weaning outcomes were recorded 24 hours after data collection. RESULTS: Participants were primarily white women and required mechanical ventilation for a mean of less than 4 days. Participants reported mild dyspnea, moderate fatigue, and high weaning self-efficacy. High PaO2, low PaCO2, stable hemodynamic status, adequate cough and swallow reflexes, no metabolic changes, and no abdominal problems were associated with complete weaning (P = .05). Subjective perceptions were associated with physiological variables but not with weaning outcomes. CONCLUSIONS: Multidimensional assessment of both primary and secondary indicators of readiness to be weaned is necessary for timely, efficient weaning from mechanical ventilation. Primary assessments include physiological variables related to gas exchange, hemodynamic status, diaphragmatic expansion, and airway clearance. Secondary assessments include perceptions related to key physiological variables. Additional research is needed to determine the predictive value of physiological variables and perceptions of dyspnea, fatigue, and self-efficacy. PMID- 12625169 TI - Oral care interventions in critical care: frequency and documentation. AB - BACKGROUND: No data have been collected to describe the products, methods, and frequency of oral care needed to reduce dental plaque, oral colonization, and ventilator-associated pneumonia in critically ill patients. OBJECTIVES: To describe the frequency of use of oral care interventions reported by nurses in several intensive care units in a large southeastern medical center. METHODS: Staff members completed a written survey describing their oral care practices, and oral care interventions were recorded from the unit's flow sheet for the previous 24 hours for all patients at 5 randomly selected times during 1 month. RESULTS: Most respondents (75%) reported providing oral care 2 or 3 times daily for nonintubated patients, and 72% reported providing care 5 times daily or more for intubated patients. However, oral care was documented on the unit's flow sheet a mean of 1.2 times per patient. Reported use of toothpaste and a toothbrush was significantly greater in nonintubated patients (P < .001), and use of a sponge toothette was significantly greater in intubated patients (P < .001). Nurses' mean rating of oral care priority was 53.9 on a 100-point scale. CONCLUSIONS: Despite evidence that they are ineffective for plaque removal, sponge toothettes remain the primary tool for oral care, especially in intubated patients in intensive care units. Nurses report frequent oral care interventions, but few are documented. Education and focus on good oral care strategies are required; nursing research to delineate the best procedure for all patients in intensive care units is needed. PMID- 12625170 TI - Advances in the understanding of clinical manifestations and therapy of severe sepsis: an update for critical care nurses. AB - Severe sepsis is a major public health concern and a burden on the healthcare system. Despite improvements in efforts to control the source of infection and increased recognition by healthcare providers of patients with the disease, the mortality rate remains unacceptably high, from 30% to 50%. The systemic inflammatory response syndrome criteria are used as diagnostic indicators of sepsis when they occur in patients with known or suspected infection. The outcome of a patient with severe sepsis is often related to the occurrence of sepsis induced multiple organ dysfunction syndrome. Multiple organ dysfunction syndrome appears to result from a cascade of organism-related factors, inflammatory mediators, endothelial injury, disturbed hemostasis, and microcirculatory abnormalities. In patients with severe sepsis, derangements of inflammation and coagulation are tightly linked. Although numerous clinical trials focused on interventions in one or the other of the inflammatory and coagulation systems failed to show reduced mortality due to sepsis, a member of a new class of drugs called "cogins" was effective. In its active form, protein C has anti inflammatory, antithrombotic, and profibrinolytic properties that can reduce organ injury associated with severe sepsis. A recombinant form of activated protein C, drotrecogin alfa (activated), significantly reduces 28-day mortality due to all causes in patients with severe sepsis and has an acceptable safety profile. This review provides an overview of severe sepsis, highlighting recent advances in treatment of the disease and the role of critical care nurses. PMID- 12625171 TI - From theory to practice: preventing pain after cardiac surgery. AB - A pain management guideline was developed at the Royal Columbian Hospital, New Westminster, British Columbia, to prevent pain after cardiac surgery. The guideline was based on a wellness model and was predicted on the World Health Organization's analgesic ladder. Patients are given nonopioids around the clock and throughout the postoperative stay and are given an opioid to prevent procedural pain and treat breakthrough pain. In an evaluation of the guideline, records from 133 cardiac surgery patients were retrospectively reviewed. The type and dose of analgesics administered for the first 6 days after surgery, the effectiveness of the pain management plan, the occurrence of adverse effects, time to extubation, and postoperative lengths of stay were determined. Ninety five percent of patients had effective pain relief. Almost all patients received acetaminophen around the clock. A total of 89% received indomethacin. All patients received opioids intermittently. Doses of opioids were converted to morphine oral equivalents, which peaked on day 1 after surgery (38 equivalents) and decreased sharply by day 2 (< 10 equivalents). Median postoperative length of stay was 5 days for patients who had bypass surgery and 6 days for patients who had valve surgery. This proactive, low-tech, low-risk, well-tolerated pain management approach is cost-effective, simple, and feasible to use. The findings support use of this approach in managing pain after cardiac surgery. PMID- 12625172 TI - Phantom shock in a patient with an implantable cardioverter defibrillator: case report. PMID- 12625173 TI - Exposure of intensive care unit nurses to nitric oxide and nitrogen dioxide during therapeutic use of inhaled nitric oxide in adults with acute respiratory distress syndrome. AB - BACKGROUND: Although low concentrations of inhaled nitric oxide may by therapeutic, both nitric oxide and its oxidation product nitrogen dioxide are potentially toxic. The threshold limits for time-weighted average concentrations of nitric oxide and nitrogen dioxide issued by the American Conference of Governmental Industrial Hygienists are 25 and 3 ppm, respectively. The concentrations of these gases in the breathing space of hospital personnel during administration of nitric oxide to adult patients have not been reported. METHODS: Air was sampled from the breathing zone of intensive care unit nurses via collar mounted tubes during the nurses' routine duties attending patients who were receiving inhaled nitric oxide at 5 or 20 ppm. The exhaust ports of the mechanical ventilators were left open to the room. Nitric oxide and nitrogen dioxide were chemically assayed as nitrite from sorbent tubes by using spectrophotometry. Ambient nitric oxide levels were measured at sequential distances from the ventilator by using chemiluminescence. RESULTS: The time weighted average concentrations of inspired gas for nurses during inhaled nitric oxide treatment were 0.45 ppm or less for nitric oxide and less than 0.29 ppm for nitrogen dioxide. Nitric oxide levels at the ventilator during delivery at 20 ppm were 9.2 ppm, but dropped off markedly beyond 0.6 m (2 ft), to a mean of about 30 ppb. CONCLUSION: Inhaled nitric oxide therapy at doses up to 20 ppm does not appear to pose a risk of excessive occupational exposure to nitric oxide or nitrogen dioxide to nurses during routine delivery of critical care. PMID- 12625174 TI - Safeguarding the patient and the profession: the value of critical care nurse certification. AB - Today's critically ill patients require heightened vigilance and extraordinarily intricate care. As skilled and responsible health professionals, the 403,000 critical care nurses in the United States must acquire the specialized knowledge and skills needed to provide this care and demonstrate their competence to the public, their employers, and their profession. Recognizing that nurses can validate specialty competence through certification, this white paper from the American Association of Critical-Care Nurses and the AACN Certification Corporation puts forth a call to action for all who can influence and will benefit from certified nurses' contribution to patient care. PMID- 12625175 TI - Reflecting on what we care about. PMID- 12625177 TI - Low amplitude QRS. PMID- 12625176 TI - Obesity and the metabolic syndrome. AB - The prevalence of marked obesity is increasing rapidly among adults and has more than doubled in 10 years. Sixty-one percent of the adult population of the United States is overweight or obese. Americans are the fattest people on earth. Paradoxically these increases in the numbers of persons who are obese or overweight have occurred during recent years when Americans have been preoccupied with numerous dietary programs, diet products, weight control, health clubs, home exercise equipment, and physical fitness videos, each "guaranteed" to bring rapid results. Overweight and obesity are also world problems. The World Health Organization estimates that 1 billion people around the world are now overweight or obese. Westernization of diets has been part of the problem. Fruits, vegetables, and whole grains are being replaced by readily accessible foods high in saturated fat, sugar, and refined carbohydrates. Since class 3 obesity (morbid or extreme obesity) is associated with the most severe health complications, the incidence of hypertension, stroke, heart disease, diabetes, and peripheral vascular disease will increase substantially in the future. Recently, obesity alone has been implicated in the development of cardiac hypertrophy and CHF. The metabolic syndrome associated with abdominal obesity, which includes insulin resistance, dyslipidemia, and elevated CRP levels, identifies subjects who have an increase in cardiovascular morbidity and mortality. Twenty to 25% of the adult population in the United States have the metabolic syndrome, and in some older groups this prevalence approaches 50%. The prevalence of overweight children in the United States has also been increasing dramatically, especially among non Hispanic blacks and Mexican-American adolescents. Overweight children usually become overweight adults. Atherosclerosis begins in childhood. The degree of atherosclerotic changes in children and young adults can be correlated with the presence of the same risk factors seen in adults. As health providers, our direction is obvious! PMID- 12625178 TI - The patient's right to choose. PMID- 12625180 TI - Inroads in implantology: managing the severely resorbed maxillary alveolus. PMID- 12625181 TI - Launch of the dental laser drill. PMID- 12625182 TI - Managed health care and dentists in the Gauteng province. AB - Managed health care (MHC) has emerged in South Africa as an alternative system to control the cost of health care. Fears, negative perceptions and much uncertainty have surrounded this issue since its introduction in South Africa. The purpose of this study was to assess the understanding, perceptions and opinions of dentists residing in the Gauteng province about the emergence, benefits, impact, and future of MHC in South Africa. Eighty-six dentists (21.5%) out of the sample population of 400 responded to the questionnaire. The majority (66.7%) of the respondents perceived MHC to be a threat to their practices, with most feeling that MHC will interfere with the doctor-patient relationship. Forty-two per cent of the respondents were of the opinion that MHC will not succeed in South Africa. The results suggest that this sample of dentists do not consider MHC to be an alternative to the existing fee-for-service system. The introduction of MHC in South Africa is mostly based on models used in the USA, which might not be appropriate in the unique South African environment. PMID- 12625183 TI - Spatial and temporal localisation of bone morphogenetic protein-3 (osteogenin) in the developing rat submandibular gland. AB - Submandibular gland morphogenesis is a highly regulated process modulated by epithelio-mesenchymal interactions. Bone morphogenetic proteins (BMPs), members of the transforming growth factor-beta (TGF-beta) superfamily, are known to be soluble mediators of epithelio-mesenchymal interactions during the development of certain organs. The aim of this study was to localise bone morphogenetic protein 3 (BMP-3 or osteogenin), spatially and temporally in the developing rat submandibular gland. Immunocytochemistry was performed on sections of the developing submandibular gland (gestation ages E14.5-E19.5). BMP-3 was localised in the extra-cellular matrix of the mesenchyme of the gland in all the stages examined. Intense staining of BMP-3 was observed in the epithelial cells of the developing end-buds between stages E14.5 and E16.5. As cytodifferentiation progressed (stage E17.5 onwards) the number of epithelial cells in the developing acini in which BMP-3 was present became markedly reduced. Similarly, the number of cells containing BMP-3 in the developing ducts decreased as duct development progressed. By stage E19.5, BMP-3 was located mainly in the immature ducts. While the effects of BMP-3 on matrix macromolecules could not be deduced from this study, its spatial and temporal location within the developing glands may indicate a role in the co-ordinated regulation of branching morphogenesis. PMID- 12625184 TI - [What do dental students at the University of Stellenbosch know of the education and functions of oral hygienists?]. AB - The purpose of this study was to determine the knowledge of dental students regarding the requirements for the course in oral hygiene at the University of Stellenbosch, the functions of oral hygienists and how they rate the necessity of the profession compared with other health care providers. Students in their second, third, fourth and fifth years of study were asked to complete a questionnaire. Although obvious gaps existed in the knowledge of a notable number of students, the majority of students in all the years showed good insight into the requirements for and contents of the course and functions of an oral hygienist. The number of second- and third-year students who gave the correct answers were consistently fewer than the senior students, indicating that when dental students come into contact with oral hygiene students in their clinical years their knowledge of the course and the profession improves. The void in the knowledge of dental students which most certainly needs correction is the limited concept some of them have about the training oral hygienists receive in periodontics (especially the preclinical students), orthodontics, medicines, legal aspects concerning dentistry, first aid and diagnostic radiology. Regarding the functions of oral hygienists, their importance was regarded by many students as comparable with that of occupational therapists, radiographers, nurses, dieticians and dental therapists, notably higher than that of dental assistants and slightly less important than that of physio-therapists. PMID- 12625185 TI - National pathfinder survey of periodontal status and treatment needs in The Gambia. AB - OBJECTIVE: The aim of the pathfinder survey was to assess the periodontal status and treatment needs in The Gambia. METHODS: A multistage stratified random sampling technique was used to obtain a sample of 1,235 (or 0.1% of the national population). The study was conducted according to the World Health Organisation's criteria (WHO, 1987). The WHO J2 software programme was used to analyse the data. RESULTS: Not more than 12% of subjects in any age group had healthy periodontal tissues. Two to three sextants were healthy among those under the age of 35 years. Shallow (4-5 mm) pockets were present in all ages. However, less than one sextant was involved among subjects younger than 25 years. Despite the high prevalence of pockets, few sextants were edentulous. The majority of subjects needed oral hygiene instructions and oral prophylaxis. Of the subjects between the ages of 8 and 29 years, 5-28% needed complex treatment in only half of a sextant. Similarly, 38% and 80% of older subjects needed complex treatment in 0.8 1.9 sextants. CONCLUSION: A national oral health plan in The Gambia should focus on health education and provision of oral prophylaxis by trained auxiliary health care workers. In conclusion, there is a need to develop a functional district oral health services system in The Gambia. PMID- 12625187 TI - Dental abscesses as a cause of 'unexplained' recurrent fever in a 9-year-old boy. PMID- 12625186 TI - Oral health care in diabetes mellitus. AB - Diabetes mellitus produces many complex changes in the lives of those affected. Elevated blood glucose levels, which may occur in the absence of symptoms, lead to late complications from tissue damage. There is an increased susceptibility to infection, poor wound healing and periodontal disease. Furthermore, chronic oral infection itself may contribute to raised blood glucose levels and hence to the later complications of diabetes. Acute infection in the oral cavity needs specific and aggressive management, just as in the acutely infected diabetic foot.' The dental team may not have made a significant contribution to the shared management of the person with diabetes in the past; however, recent findings suggest that the dental team may contribute greatly to the shared care of diabetes with the diabetic team itself, and regular liaison is strongly recommended. PMID- 12625188 TI - Conscious sedation. PMID- 12625190 TI - Direction of hand preference: the connection with speech and the influence of familial handedness. AB - Taking familial handedness into account, right- and left-handers with differing degrees of hand preference were monaurally presented with verbal stimuli (CVs) to which they responded using their right and left hands at separate times. This reaction time design was used to ascertain the relative cognitive functioning capacity of each hemisphere for verbal processing. However, the results disclosed more about the determination of direction of hand preference than about cognitive processing, per se. It was found that in both strong right-handers and strong left-handers with an incongruent hand preference (i.e., own handedness incongruent with family history of handedness) direction of hand preference is the result of suppression of the nonpreferred hand in the left hemisphere. Strong right- and strong left-handers with a congruent hand preference (i.e., hand preference congruent with family history of handedness) appear to have a more direct hand preference-left hemisphere mechanism. The findings of the present study are used to form part of a new theory of hand preference determination. PMID- 12625191 TI - EEG asymmetry responses to lavender and rosemary aromas in adults and infants. AB - Frontal EEG asymmetry shifting from baseline was examined in adults and infants exposed to lavender and rosemary by reanalyzing previously published data, using techniques different from those utilized in the original two studies. Results from Study 1 on 39 adults revealed significant EEG shifting in the lavender group, with greater relative left frontal EEG activation (associated with greater approach behavior and less depressed affect). The participants in the two aroma groups were further grouped by those with greater baseline, relative to left frontal EEG activation, versus those with a greater baseline, relative to right frontal activation. Collapsing across aroma groups, those with greater baseline, relative to right frontal activation, shifted left during the aroma. Those with greater baseline relative to left frontal activation did not change. In the rosemary group, those with greater baseline relative to right frontal EEG activation shifted left during the aroma, while those with greater baselines relative to left frontal EEG activation shifted right. In the lavender group, those with greater baselines relative to right frontal baseline EEG activation shifted left, but those with greater baselines relative to left baseline did not shift. Study 2 on 27 full-term newborns revealed no significant shifts in asymmetry in either aroma group. However, when the aroma groups were collapsed, the right frontal EEG group exhibited significant shifting relative to left frontal EEG activation. This finding was similar to the adult findings, suggesting that either lavender or rosemary may induce left frontal EEG shifting in adults and infants who show greater baselines relative to right frontal EEG activation. PMID- 12625192 TI - The effect of different hypertension models on visual evoked potentials. AB - Even though there is an abundance of information about the complications of hypertension, studies of its influence on visual evoked potentials (VEPs) are rare. In previous studies, it was pointed out that hypertension induces changes on VEPs. However, it has not yet been clarified which models of hypertension are more effective on VEPs. The aim of this study was to investigate this subject in rats. Animals were divided equally into six groups: control group (C), sham operated (Sham), two kidney-one clip (2K-1C), one kidney-one clip (1K-1C), deoxycorticosterone-salt (DOCA), and N-omega-nitro-L-arginine-methyl ester (L NAME) groups. Mean arterial pressure was significantly higher in four hypertensive groups compared with control and sham groups, but there were no significant differences either among hypertensive groups or between sham and control groups. At the end of the experimental period, flash visual evoked potentials were recorded. The mean latencies of P1, N1, P2, N2, and P3 components were significantly prolonged in all hypertensive groups compared with the control and sham groups. The mean latencies of all VEPs components in the L-NAME group were longer than in the other hypertensive groups. Thiobarbituric acid-reactive substances (TBARS) were determined as an indicator of lipid peroxidation. Our data showed that hypertension caused a significant increase of lipid peroxidation in brain and retinal tissues. Additionally, plasma renin activity (PRA) was highest in the 2K-1C group and lowest in the DOCA group. PMID- 12625194 TI - Measuring various sizes of H-reflex while monitoring the stimulus condition. AB - The purpose of this study was to assess the usefulness of a new technique that measured various sizes of the soleus H-reflex, while monitoring the stimulus condition. Eight healthy volunteers participated in this experiment. In the new technique, an above-motor-threshold conditioning stimulus was given to the tibial nerve 10-12 ms after a below-motor-threshold test stimulus. The conditioning stimulus evoked a direct M-wave, which was followed by a test-stimulus-evoked H reflex. This reflex was followed by a conditioning stimulus-evoked H-reflex. The amount of the voluntary-contraction-induced facilitation of the H-reflex was similar for both the new technique and conventional technique, in which an above motor-threshold test stimulus was given without a conditioning stimulus. Using the new technique, we found that the amount of facilitation increased linearly with the size of the test H-reflex. This technique allows us to evoke various sizes of H-reflex while monitoring a stimulus condition, and is useful for measuring H-reflexes during voluntary movement. PMID- 12625193 TI - A simple, inexpensive, and precise spectrophotometric method for evaluating the concentration of ascorbic acid in CSF samples: data from different neurological pathologies. AB - A method is presented for evaluating the concentration of ascorbic acid in 10 microliters samples of cerebrospinal fluid (CSF) with just ultraviolet (UV) scanning in the 220-300 nm range. The method assumes that the two main UV absorbing CSF components are proteins (with a peak at 280 nm) and ascorbic acid (with a peak at 264 nm). On this basis, the absorbances at those wavelengths are the starting points for a calculation that evaluates the sheer contribution of ascorbic acid to the absorbance at 264 nm. The rapidity of the UV analysis (just a few min), together with the precaution of keeping the samples under argon, along the short preparatory procedure, is especially suitable in the analysis of an air labile substance, such as ascorbic acid. The results of the UV procedure have been checked with parallel HPLC determinations for 19 test CSF samples and the results have shown an excellent correspondence. Finally, data are presented about the evaluation of ascorbic acid concentrations in CSF samples from various neurological pathologies in comparison with normal cases. These data show the interesting result of a significant reduction in the average ascorbic acid CSF level in patients suffering from Alzheimer's disease. However, a decrease in that parameter is found also for a few other pathologies. PMID- 12625195 TI - Correlations among behavioral measures of orbitofrontal function. AB - Orbitofrontal cortex plays an important role in guiding behavior based on emotional input and rewards. It receives multimodal higher order sensory information and mediates reinforcement for primary as well as secondary or conditioned forms of reinforcement (e.g., monetary rewards). Several behavioral measures have demonstrated putative sensitivity to orbitofrontal function: smell identification, delayed alternation, and response inhibition (go/no-go and antisaccades). A correlational study of performances on these tasks by healthy controls demonstrated some interrelationships: go/no-go correlated with antisaccades, left nostril smell correlated with go/no-go, and right nostril smell correlated with delayed alternation. Further analysis showed that certain smells correlated with tasks more strongly than others. Given the diversity of these tasks, it is suggested that these intercorrelations results from at least the partial overlap of the neural substrates for olfactory identification and tasks requiring response modification based on reinforcement contingencies. PMID- 12625196 TI - No effect of long-term valproate therapy on thyroid and parathyroid functions in children. AB - In this study, we studied serum calcium, phosphorus, alkaline phosphatase, thyroid hormones (total thyroxine, free thyroxine, thyroid-stimulating hormone), parathyroid hormone, and osteocalcine levels in children with epilepsy who had been receiving long-term valproate (VPA) therapy in order to determine whether there was any effect of VPA therapy on these hormones. The study included 31 patients with epilepsy receiving VPA and 22 healthy age-matched controls. The age ranged from 15 months to 16 years and 18 months to 17 years in the study and control group, respectively. The duration of VPA use was between 12 months and 5 years (1.93 +/- 1.90 years). When comparing the results, we did not find any significant difference in any of the parameters, including serum calcium, phosphorus, alkaline phosphatase, osteocalcine, and thyroid and parathyroid hormone levels, between the study and control group. We suggest that VPA can safely be used with regard to thyroid and parathyroid dysfunction in childhood epilepsy. PMID- 12625197 TI - The efficacy of neuropsychological symptom inventory in the differential diagnosis of medical, psychiatric, and malingering patients. AB - The present study was conducted to examine patient responses to the Neuropsychological Symptom Inventory (NSI) and attempted to discriminate among medical, psychiatric, and simulated malingering patients. Results indicated that the NSI was able to significantly discriminate malingered responses from medical and psychiatric patient responses. However, applying a lie scale derived from previous research with the NSI did not allow discrimination between the malingered group and the psychiatric patients. Using the factors of the NSI derived from earlier research offered both greater detection of malingerers and enhanced evaluation of symptom profiles of medical and psychiatric patients. The NSI provides an efficient screen for exaggerated symptoms, as well as an indication of the level of general neuropsychological functioning of the patient when included in a neuropsychological evaluation. PMID- 12625198 TI - Conflicting professional values in medical education. PMID- 12625199 TI - The best lack all conviction: biomedical ethics, professionalism, and social responsibility. PMID- 12625200 TI - Understanding, being, and doing: medical ethics in medical education. PMID- 12625201 TI - The moral orientations of justice and care among young physicians. PMID- 12625202 TI - Bioethics and graduate medical education: the great match. PMID- 12625203 TI - Justice in residency placement: is the match system an offense to the values of medicine? PMID- 12625205 TI - Helping residents live at risk. PMID- 12625206 TI - Workplace bullying, psychological distress, and job satisfaction in junior doctors. PMID- 12625207 TI - Lessons from a bygone medical ethics program. PMID- 12625208 TI - CQ sources/bibliography. PMID- 12625209 TI - Response to "Do genetic relationships create moral obligations in organ transplantation?" By Walter Glannon and Lainie Friedman Ross (CQ Vol 11, No 2). PMID- 12625211 TI - Unethical author attribution. PMID- 12625210 TI - Organizational ethics in residency training: moral conflict with supervising physicians. PMID- 12625212 TI - Recap of FDA product approvals--2002. PMID- 12625213 TI - Clinicians, students fill health services gap for Houston's homeless. PMID- 12625214 TI - Improving care and reducing costs through partnerships between clinical and financial leaders. PMID- 12625215 TI - Programs established by FDA to expedite patient access to medications. PMID- 12625216 TI - Biological response modifiers in the management of rheumatoid arthritis. AB - The management of rheumatoid arthritis (RA) with biological response modifiers (BRMs) is reviewed. RA, an autoimmune disorder affecting 1-2% of the world's population, is characterized by inflammation of synovial tissues, joint swelling, stiffness, and pain that may progress to joint erosion. There is strong evidence that inflammatory mediators, such as tissue necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1), play a critical role in the pathogenesis of this disorder. IL-1-receptor antagonist (IL-1Ra) is produced in healthy subjects and helps to protect against the adverse effects associated with IL-1 overexpression. Administration of IL-1Ra or similar agents may reduce the effects of IL-1 and ameliorate inflammatory conditions. Traditional treatment of RA has been based on symptomatic management with non-steroidal antiinflammatory drugs, disease modifying antirheumatic drugs, and corticosteroids, each of which has substantial drawbacks in terms of effectiveness or adverse effects. Newer therapeutic strategies for blocking the biological effects of inflammatory cytokines include antibodies directed against TNF (e.g., infliximab), soluble receptors (e.g., etanercept) and receptor antagonists to IL-1 (anakinra) [corrected]. Clinical trials indicate that these BRMs may be more effective than traditional agents because they are able to alter joint remodeling in addition to attenuating symptoms. Anti-TNF therapies may be associated with increased risk for infections, sepsis, tuberculosis reactivation, demyelination disorders, and blood dyscrasias; anakinra appears to be safer. Combination therapy with BRMs may be more appropriate for RA than monotherapy. The role of BRMs in the treatment of RA will evolve as investigators learn more about the drugs and the disorder. PMID- 12625217 TI - Bitter melon (Momordica charantia): a review of efficacy and safety. AB - The pharmacology, clinical efficacy, adverse effects, drug interactions, and place in therapy of bitter melon are described. Bitter melon (Momordica charantia) is an alternative therapy that has primarily been used for lowering blood glucose levels in patients with diabetes mellitus. Components of bitter melon extract appear to have structural similarities to animal insulin. Antiviral and antineoplastic activities have also been reported in vitro. Four clinical trials found bitter melon juice, fruit, and dried powder to have a moderate hypoglycemic effect. These studies were small and were not randomized or double blind, however. Reported adverse effects of bitter melon include hypoglycemic coma and convulsions in children, reduced fertility in mice, a favism-like syndrome, increases in gamma-glutamyltransferase and alkaline phosphatase levels in animals, and headaches. Bitter melon may have additive effects when taken with other glucose-lowering agents. Adequately powered, randomized, placebo-controlled trials are needed to properly assess safety and efficacy before bitter melon can be routinely recommended. Bitter melon may have hypoglycemic effects, but data are not sufficient to recommend its use in the absence of careful supervision and monitoring. PMID- 12625218 TI - Detection of medication nonadherence through review of pharmacy claims data. AB - The utility of pharmacy claims data in detecting improper medication use, medication changes, and formulary adherence was assessed. Pharmacists provided six months of pharmacy claims data to primary care physicians for patients on the day of their scheduled clinic appointments. Similar data, not supplied to physicians, were generated for a matched population of patients on the day of their scheduled clinic appointments. A blinded pharmacist abstracter reviewed dictated office notes for both control and treatment groups. Medication adherence and medication changes were assessed by the abstracter as well as the difference in total medication costs for each of the groups before and after the pharmacy claims data were supplied. Surveys were distributed to physicians to determine whether pharmaceutical care was improved by the pharmacy claims data. In the treatment group, physicians detected medication nonadherence in 30.5% of their patients, while the abstracter noted nonadherence in 58.1% of patients. Physicians failed to detect any nonadherence in the control group, but the abstractor detected nonadherence in 57.1% of these patients. Changes in medication regimens occurred more often in the treatment group (p < 0.001). The mean percentage of patients switched to formulary agents significantly differed between the treatment and control groups (27.7% versus 0.0%, respectively) (p < 0.001). There were no differences in median drug costs for either group before or after the pharmacy claims data were provided. Provision of pharmacy claims data to physicians helped them detect medication nonadherence, evaluate therapeutic duplication or omissions, increase formulary use, and reduce the time required to obtain an accurate medication history. PMID- 12625219 TI - Hospital policies regarding herbal medicines. AB - The prevalence and content of herbal policies and herbal formularies in the hospital setting were studied. Drug information centers affiliated with hospital pharmacies were surveyed by telephone. Hospitals with policies on herbal products were asked to provide detailed information about the policies. Of 70 hospitals included in the analysis, 53 (76%) had policies and procedures on the use of herbal products. Three hospitals (4%) reported having an herbal formulary. A majority of the existing policies required a physician order for an herbal product to be used inhouse. Many of the policies also required pharmacists to verify labeled product ingredients. Product administration, patient consent, and drug interaction screenings were addressed by some hospitals. Most hospitals reported having policies on the use of herbal products, but herbal formularies were rare. PMID- 12625220 TI - Reducing claims for prescription drugs with a high potential for abuse. PMID- 12625221 TI - Sudden hearing loss associated with ephedra use. PMID- 12625222 TI - Cost-benefit analysis of a pharmacy-managed medication assistance program for hospitalized indigent patients. PMID- 12625223 TI - Distinction between innovative therapy and research. PMID- 12625224 TI - 2002 ASHP National Residency Preceptors Conference celebrates 40 years of programs, emphasizes caring. PMID- 12625225 TI - Impact of aspirin on the gastrointestinal-sparing effects of cyclooxygenase-2 inhibitors. PMID- 12625226 TI - Quantifiable impact of the contract for health and wellness: health behaviors, health care costs, disability, and workers' compensation. AB - Current literature about the long-term impacts of corporate health and wellness programs has brought to light new evidence about the cost savings associated with health-promotion interventions. A critical element in these initiatives is attracting the participation of employees at risk for high benefits use. This study presents evidence that suggests accomplishing this task has economic savings implications to large employers. A health and wellness intervention program offered at GlaxoSmithKline, entitled the Contract for Health and Wellness, is examined. Focusing on a group of 6049 employees, the study examines the impact on health behaviors and on integrated health benefits use of this continuously employed population from 1996 to 2000. Total benefits costs are examined for participants and nonparticipants, and the annual savings associated with the isolated impact of the program are, on average, $613 per participant. Reductions in disability costs accounted for the majority of these savings. PMID- 12625227 TI - Association between human paraoxonase gene polymorphism and chronic symptoms in pesticide-exposed workers. AB - Pesticides, such as parathion, are metabolized by cytochrome p-450 system to paraoxon, which is a potent cholinesterase inhibitor. Paraoxonase (PON) catalyzes the hydrolysis of these toxic metabolites and protects against pesticide toxicity. A glutamine/arginine (Gln/Arg) polymorphism at amino acid position 192 of PON has been described. The Arg/Arg genotype is associated with higher serum paraoxonase activity compared to Gln/Gln. The Arg/Gln genotype is associated with intermediate serum PON activity. The potential association between PON genotype and symptoms of chronic pesticide toxicity was examined among 100 farm workers. As part of a cross-sectional study of pesticide toxicity among mixed-race farm workers in the Western Cape. South Africa, 100 farm workers were genotyped for polymorphism of the paraoxonase gene at amino acid position 192. Subjects with two or more of the following symptoms were considered to have evidence of chronic toxicity: abdominal pain, nausea, rhinorrhea, dizziness, headache, somnolence, fatigue, gait disturbance, limb numbness, paresthesias, limb pain, or limb weakness. In multivariable logistic regression analysis, the independent predictors of chronic toxicity were previous history of head trauma resulting in loss of consciousness (OR 2.8, 95% CI = 1.7-6.7), having worked as a pesticide applicator (OR 5.4, 95% CI = 3.2-8.9), and having one of the two "slow metabolism" (Gln/Gln or Gln/Arg) genotypes (OR 2.9, 95% CI = 1.7-6.9). Furthermore, the prevalence of chronic toxicity increased in a stepwise fashion from 15% among pesticide nonapplicators with a "fast metabolism" (Arg/Arg) genotype, to 42.9% among pesticide nonapplicators with "slow metabolism" (Gln/Gln or Gln/Arg) genotypes, to 58.8% among pesticide applicators with "fast metabolism" genotype, and 75.0% among pesticide applicators with "slow metabolism" genotypes (P = 0.001). Age, number of years on the job, smoking history, alcohol history, education level, plasma or red blood cell cholinesterase level, or previous history of acute organophosphate poisoning were not statistically significant predictors of chronic toxicity. The PON genotype is an important determinant of a farmworker's susceptibility to chronic pesticide poisoning. PMID- 12625228 TI - Job insecurity as a risk factor for common infections and health complaints. AB - The aim of this study was to investigate the cross-sectional and longitudinal effect of job insecurity on common infections and health complaints. Self administered questionnaire data from the Maastricht Cohort Study (n = 12, 140) were used. We used Generalized Estimating Equations analyses to calculate pooled odds ratios (ORs) and 95% confidence intervals. A cross-sectional relationship between job insecurity and common infections or health complaints was found. For the longitudinal relationship, the largest effect was found for flu-like illness (OR 1.39; CI = 1.22-1.57) and health complaints (OR 1.51; CI = 1.39-1.64). We additionally corrected for health behavior, the presence of a long-standing illness, and work-related demands, resulting in lower ORs. An increase in common infections or health complaints has a substantial impact on employee well-being and may result in economic consequences for the company. PMID- 12625229 TI - A triangulation approach to historical exposure assessment for the carbon black industry. AB - The determination of cumulative exposures for individual workers is necessary for research and practice of occupational health and hygiene. Reconstruction of exposures for a study of respiratory morbidity was needed to study the effects of exposure to carbon black production. Approximately 15,800 exposure estimates were needed. There were 22 plants, a 40-year time span, six job categories, and three types of dust-exposure metrics (respirable, inhalable, and "total" dust). Three information sources were used: 1) Industrial hygiene air level measurements where available (several industry-wide surveys had been conducted). 2) A formal process survey identifying specific dates and types of process and control changes. 3) An Historical Relative Exposure Rating Scale; plant health and safety personnel used this spreadsheet-based rating scheme to quantify exposures before and between years of actual measurement relative to a reference year in which measurements were available. A job-exposure matrix was calculated by integrating these three methods. Linear scaling factors were identified to interconvert geometric to arithmetic means and to interconvert total and inhalable dust. Individual worker cumulative exposures were then calculated based upon job histories linked with the job-exposure matrix. The nine-step process for integrating all available relevant data was effective in estimating the exposures for each of the cells of the job-exposure matrix. Among the 1680 workers participating, the mean cumulative inhalable dust exposure was 48.4 mg-years/m3. Early years contribute disproportionately to the cumulative exposures of individuals since levels have declined significantly over time. The use of multiple sources of information, including a relative exposure rating instrument, significantly facilitates reconstruction of historical exposures. Inadequate adjustment for temporal trends can lead to underestimation of cumulative exposures and significantly affect estimation of dose-effect relationships. These methods are applicable to other situations requiring estimation of cumulative exposure with sparse industrial hygiene data in early years. PMID- 12625230 TI - Effect of carbon black exposure on respiratory function and symptoms. AB - Carbon black is a widely used pigment and filler. Some, but not all, previous studies have suggested an effect of long-term exposure upon the lungs. Carbon black production facility employees (1755) participated in the third round of the industry-wide medical surveillance testing. They were employed in 22 North American plants. Spirometry and a systematically administered questionnaire were included in the year 2000 round of the industry-wide medical surveillance program. Industrial hygiene data from an industry-wide survey in 2000-2001, as well as all available exposure assessment data collected since 1979, were integrated with process questionnaires and exposure rating questionnaires completed by plant personnel. Analyses included multiple linear regression and categorical data analyses. Multiple regression analyses showed statistically significant, consistent relationships between cumulative exposure and small reductions in forced expiratory volume in 1 second (FEV1) but not with other spirometry parameters. The estimated slopes were -2 mL FEV1 per mg-year/m3 of cumulative 'total' dust exposure and -0.7 mL FEV1 per mg-year/m3 of cumulative exposure for the inhalable fraction. In addition, heavy cumulative exposures were associated with a small increase in chronic bronchitis in nonsmokers. Recent exposures, typically much lower than in the past, were not demonstrated to be associated with these effects. Consistent with good occupational hygiene practice for any contaminant, workplace exposures to carbon black should be controlled to lowest practical levels. PMID- 12625231 TI - The World Health Organization Health and Work Performance Questionnaire (HPQ). AB - This report describes the World Health Organization Health and Work Performance Questionnaire (HPQ), a self-report instrument designed to estimate the workplace costs of health problems in terms of reduced job performance, sickness absence, and work-related accidents-injuries. Calibration data are presented on the relationship between individual-level HPQ reports and archival measures of work performance and absenteeism obtained from employer archives in four groups: airline reservation agents (n = 441), customer service representatives (n = 505), automobile company executives (n = 554), and railroad engineers (n = 850). Good concordance is found between the HPQ and the archival measures in all four occupations. The paper closes with a brief discussion of the calibration methodology used to monetize HPQ reports and of future directions in substantive research based on the HPQ. PMID- 12625232 TI - Organizational climate, perceived work-related symptoms and sickness absence: a population-based survey. AB - Very few reports have been published on organizational climate, health, and sickness absence in a representative sample of the entire workforce. The aim of this study was to determine how the perceived organizational climate of a workplace is related with work-related symptoms and sickness absence and how these factors vary according to sociodemographic and work-related characteristics. Data were collected in computer-assisted telephone interviews of a random sample of 4209 currently employed Finns drawn from the population register. A tense and prejudiced climate was associated with a higher risk of work-related symptoms than a relaxed and supportive climate (odds ratio [OR] 3.0 (95% CI = 2.4-3.7). The corresponding ORs were 4.3 (95% CI = 3.3-5.6) for psychological symptoms, 1.6 (95% CI = 1.2-2.0) for musculoskeletal symptoms, and 1.6 (95% CI = 1.3-2.1) for more than the average number of sick-leave days. Part of the impact of organizational climate on sickness absence is not caused by an increase in work-related symptoms. Thus, organizational climate seems not only to correspond with organizational practices and leadership but also occupational health. Organizational climate could be used as a research tool in attempts to reduce work-related ill health and sickness absenteeism. PMID- 12625233 TI - A petroleum company's experience in implementing a comprehensive medical fitness for duty program for professional truck drivers. AB - Aggregate health risk appraisal data from Chevron Texaco's truck driver workforce revealed that approximately 50% of the truck drivers who completed a health risk appraisal were found to be at risk for a back injury. Lost time records revealed that this population had 65% more lost workdays attributed to cumulative trauma injuries compared with acute trauma injuries. Chevron Texaco addressed the issue by implementing a medical fitness for duty program consisting of three components: physical examination (physical examination mandated by the Department of Transportation and a functional capacity evaluation), education (on safe body mechanics), and physical fitness. After a 1-year development period, the fitness for duty program was implemented for Chevron Texaco's professional truck drivers in North America. In the first year of the program, 109 functional capacity evaluations were completed by trained clinicians, and 88% of candidates were found "able to work without restrictions," whereas 6% were found "able to work with caution." The article describes the program results to date, and also describes the follow-up program for drivers found to be at risk for a back injury. PMID- 12625234 TI - Hazardous ammonia releases: public health consequences and risk factors for evacuation and injury, United States, 1993-1998. AB - Unplanned releases of ammonia lead more often to evacuation and injury than releases of other chemicals, but few studies have systematically investigated ammonia releases. We analyzed Hazardous Substances Emergency Events Surveillance system data for 1993-1998. Evacuation of a total of at least 40,680 persons resulted from 537 ammonia releases, and 248 ammonia releases led to injury of 1434 persons. Equipment failure and operator error were cited as factors contributing to ammonia releases 90% of the time. Eighty-seven percent of releases occurred at fixed facilities. Risk factors for evacuation and injury differed between the food-manufacturing industry and other industries. Indoor release was a consistent risk factor, whereas quantity of ammonia released was not always a risk factor. Preventive maintenance and worker training may be effective tools to reduce the burdens of hazardous ammonia releases. PMID- 12625235 TI - [Schizophrenia and perspective taking: a comparison of schizophrenic and transient psychotic disorder patients]. AB - The purpose of the present study was to investigate the ability of schizophrenic patients to take another person's perspective. Two perspective taking tasks were examined: perceptual and affective perspective taking. Three groups participated in the study: 22 schizophrenics, 11 transient psychotic disorder [TPD] patients, and 20 normal students. Both schizophrenic and TPD patients scored significantly lower in the both perspective taking tasks than the normal controls. The affective perspective taking scores of the two patients groups were compared with those of six year-old children. Although schizophrenic and TPD patients were thought to be able to infer another's affective state correctly, they sometimes made errors in the task, because they failed to control for their own affect. Thus, their response tendency was different from the normal children of the age. The two patients groups appeared to have a common specific difficulty in inferring another's emotions. PMID- 12625236 TI - [How does the emotionality of the witnessed event affect the source monitoring ability?]. AB - This study investigated if emotionality affects the occurrence of source misattribution, which seems to be the most valid mechanism of misinformation effect. A half of 120 participants saw an emotionally stressful videotaped movie, and the other half a neutral one. Then, they answered the questionnaire, which included misinformation about the details of the movie. After 15 minutes of 2 days, they answered the source monitoring test, which required them to select the source of the memory of details that had been presented in the movie and/or in the questionnaire, or not either of them. Several patterns of differences in test performance appeared between the emotional condition and the neutral condition. These differences suggest that the ability for source monitoring declined more saliently with time in the emotional condition than in the neutral condition, and that memories about visually central details are less vulnerable to misinformation effect with 15 minutes delay in the emotional condition than in the neutral condition. PMID- 12625237 TI - [Interaural attention switching suppresses N400 attenuation with spoken word repetition]. AB - Event-related brain potentials were recorded from participants who listened to two pairs of words (and occasionally nonwords) simultaneously presented to both ears with stimulus onset asynchrony (SOA) of 1 s on a trial. When participants were instructed to fixate their attention to one ear and detect nonwords, the N400 attenuation was observed for a word repeated in the attended ear. The requirement of interaural attention switching between the first and second presentations of word pairs, however, suppressed the repetition effect on N400, even when the first and repeated presentations of a word were attended. The suppression of N400 attenuation was interpreted to indicate that attention switching interferes with the immediate availability of the decision for the first presentation in working memory. PMID- 12625238 TI - [Black sheep effect and ingroup favoritism in social identity perspective]. AB - Black sheep effect (Marques, Yzerbyt, & Leyens, 1988) and ingroup favoritism (Tajfel, Billig, Bundy, & Flament, 1971) appear to contradict each other with respect to the evaluation of inferior ingroup members. In addition to examining the relationship between the two phenomena, this study also examined the effect of ingroup identification on them. Sixty-one female student-nurses participated in the study. To measure black sheep effect, superior and inferior ingroup and outgroup members were rated on 20 adjectives. Distribution matrices (Tajfel et al., 1971) were used to measure ingroup favoritism. The participants were median split according to their level of group identification. Black sheep effect was found only among high-identification participants, who also showed ingroup favoritism. Ingroup identification had a positive correlation with both ingroup favoritism and black sheep effect. Overall, ingroup favoritism had a positive correlation with the evaluation of superior members, while it had a negative correlation with that of inferior ingroup members. PMID- 12625239 TI - [Negative bias on self-referent processing in depression: focused on mood congruent effects]. AB - The purpose of this study was to investigate negative bias on self-referent processing in depression, focused on the mood congruent effects in a natural depressed state and an experimentally induced transient depressed mood state. In Experiment 1, autobiographical memories and self-relevant ratings of personality trait words were examined in a natural depressed state or non-depressed state, which were measured by Beck Depression Inventory. Results revealed the mood congruent effects on both tasks. In Experiment 2, the same tasks as Experiment 1 were conducted in a transient depressed mood state or non-depressed mood state, which were induced through listening music. Unlike Experiment 1, there were no effects in both tasks, and a positive bias was observed in both mood states. It was suggested that transient mood state did not bias self-referent processing in depression, and Beck's schema hypothesis was supported. PMID- 12625240 TI - [Mood effects on the impression formation of typical and atypical targets]. AB - In this study, we investigated mood effects on impression formation of typical and atypical target persons, in an attempt to replicate the findings reported by Forgas (1992a). Listening to a particular piece of music, participants were first induced into positive, negative, or neutral mood. Then, they read statements, in which typical and atypical targets were described with positive and negative personality traits. While reading the statements, they were asked to form an impression of the target, and evaluate him/her. After the impression formation task, they were given ten minutes for an incidental free recall test. Results showed that mood congruent memory effect was found regardless of the targets. On the contrary, mood congruent judgment effect in positive mood was found only for typical targets. It was suggested that mood effects depend on the kind of information processing strategies triggered by typical and atypical targets. PMID- 12625241 TI - [The link between positive-negative appraisals of prebereavement and mental health after spousal loss]. AB - The aim of this study is to investigate the link between the cognitive appraisals of prebereavement and mental health after spousal loss. One hundred and twenty widow(er)s answered questionnaires concerning appraisals of prebereavement (Scale for Appraisals of Prebereavement: SAP) and mental health (General Health Questionnaire Japanese version, 28 items version: GHQ-28). As a result of factor analysis with promax rotation, it was revealed that SAP had four main factors: "Human relationship with the deceased," "Life of the deceased," "End of the deceased," and "Hospital staff and care." Age of the subjects showed positive correlations with both "Total appraisal of prebereavement" and "Life of the decreased." It was shown that the SAP as a whole had very little relationship with each subscales of GHQ-28. The psychiatrically high-risk widow(er)s have more positive appraisals on both "Total appraisal of prebereavement" and "Life of the deceased." The implications of these findings for family care and bereavement care were discussed. PMID- 12625242 TI - [Relationship with husband and child, interpersonal attitude, and personal development as mother]. AB - The present study examined how relationship with husband and child and interpersonal attitude affected a women's personal development as mother. One hundred forty-two mothers with a 3- to 5-year-old child completed a questionnaire that consisted of three scales: Parent Development Scale (Kashiwagi & Wakamatu, 1994); Husband and Children Attachment Scale (Ohinata, 1988); and Attachment Style Scale (Takuma & Toda, 1988). The last was used to measure the person's basic interpersonal attitude. Results indicated first that the person's close relationship with her child had an influence over various aspects of development as mother. Second, strong attachment to her husband seemed to make the person more active in her life. Third, the interpersonal attitude appeared to be related with the person's flexibility. PMID- 12625243 TI - [Philadelphia chromosome]. PMID- 12625244 TI - [The mystery of histone H1]. PMID- 12625245 TI - [Prion (PSI) and its effect on termination of translation in Saccharomyces cerevisiae]. PMID- 12625246 TI - [Gene expression silencing and new tools in plant molecular biology]. PMID- 12625247 TI - [Participation of aminoacyl-tRNA synthetases in cellular processes]. PMID- 12625248 TI - [Depletion of cholesterol for signal transduction from endoplasmic reticulum to the cell nucleus--the proteolytic cleavage of transcription factors from the ER membranes]. PMID- 12625249 TI - [Role of cell membrane lipids in multidrug resistance and its modulation]. PMID- 12625250 TI - [Nucleotide receptors]. PMID- 12625251 TI - [Adenosine--neurotransmitter and neuromodulator in the central nervous system]. PMID- 12625253 TI - HIPAA compliance deadlines are fast approaching! AB - The 1996 Health Insurance Portability and Accountability Act is a broad and complex law that sets out requirements in areas ranging from uniform data management to the privacy and security of health care information. No home care or hospice provider will be compliant with HIPAA without changes in their operation. The author reviews the entities affected by HIPAA, provides the timetable for the law's implementation, and sorts through the wide-ranging security and privacy standards home care providers will have to incorporate into their operations. PMID- 12625254 TI - HIPAA practical strategies & issues to compliance. AB - Although the rules for the Health Insurance Portability and Accountability Act are still undergoing continuous interpretation, as the first major compliance date approaches there are several steps that home health and hospice agencies can take now to ensure that they are ready for the April 2003 deadline. The author reviews how agencies can prepare their organization and staff for enforcement of the new privacy regulations. PMID- 12625255 TI - Implementing technological safeguards to ensure patient privacy. AB - Computers play an increasingly important role in the storage and retrieval of patient data. Internet attack incidents are growing at the astronomical rate of 64 percent a year, and some estimates state that employees are responsible for more than 80 percent of all security breaches. The information security regulations of the Health Insurance Portability and Accountability Act provide an impetus for health care providers to implement processes that ensure the safety and confidentiality of patient information. PMID- 12625256 TI - The technology-privacy conundrum. HIPAA perspectives for telehome care providers. AB - The implementation of Health Insurance Portability and Accountability Act privacy and security provisions does not need to be a cause for panic among health care organizations. A common sense approach that stresses documentation of policies and procedures for communication using technology, such as in the case of providing telehome care, takes agencies a long way towards HIPAA compliance. PMID- 12625257 TI - Personal care services: making the most of limited Medicaid funds. PMID- 12625258 TI - Home care heroes from 9/11. PMID- 12625259 TI - Home care at the crossroads. PMID- 12625260 TI - A chance to make it better. PMID- 12625261 TI - Perspectives. Bush health agenda: less than meets the eye. PMID- 12625263 TI - All-in-one surgery for heart failure. New techniques, materials and multitasking offer extra years of life. PMID- 12625262 TI - Space life sciences: extraterrestrial organic chemistry, UV radiation on biological evolution, and planetary protection. Proceedings of the F3.4(1)-B0.8, F3.1 and F3.5-PPP1 Symposia of COSPAR Scientific Commission F which were held during the Thirty-third COSPAR Scientific Assembly, Warsaw, Poland, July, 2000. PMID- 12625264 TI - Emboli-catching "umbrella" protects high-risk patients. PMID- 12625265 TI - The danger of not dialing 911. PMID- 12625266 TI - Antiplatelet drug works better if used longer. PMID- 12625268 TI - Heart protection without HRT. Options abound in the wake of negative news for women. PMID- 12625267 TI - The fewer the teeth, the greater the stroke risk. PMID- 12625270 TI - Study supports Atkins diet; heart association is skeptical. PMID- 12625269 TI - Net benefit: fish for your health. If you can't tolerate seafood, consider fish oil supplements. PMID- 12625271 TI - When the subject is sex, shhh... Patients' questions often go unanswered, a survey reveals. PMID- 12625272 TI - Ask the doctors. I live in a city with high air pollution. Is this a risk factor for heart disease? PMID- 12625273 TI - Ask the doctors. I have prominent varicose veins on both legs. They hurt, and I don't like how they look. How can I get rid of them? PMID- 12625274 TI - Ask the doctors. My doctor said recently that I have pre-diabetes. What does that mean? PMID- 12625275 TI - Sciatica. Lower back pain with many causes. PMID- 12625276 TI - Health tips. Tennis elbow and golfer's elbow. PMID- 12625277 TI - Test for acid reflux more convenient than previous tests. PMID- 12625278 TI - New treatment for pain caused by bone cancer. PMID- 12625279 TI - Managing your weight. A moving experience. PMID- 12625280 TI - Seasonal depression. A matter of light. PMID- 12625281 TI - Improving bone density. Tips may help. PMID- 12625282 TI - What exactly is a pterygium? My ophthalmologist says I have the beginnings of one. PMID- 12625283 TI - I've read that taking a chromium supplement can help increase metabolism. Is this true? Are there side effects? PMID- 12625284 TI - FDA approves generic drug for persistent heartburn. PMID- 12625285 TI - FDA approves OTC Claritin. PMID- 12625286 TI - FDA approves new HIV test kit. PMID- 12625287 TI - Dietary supplements seized after autism claims. PMID- 12625288 TI - Health advisory on sprouts. PMID- 12625290 TI - Heart pump approved for permanent use. PMID- 12625289 TI - New lab test cleared for strep in pregnant women. PMID- 12625291 TI - Cyber warnings for herbal products promoter. PMID- 12625292 TI - New warnings for Bextra. PMID- 12625293 TI - Drugs approved for opiate dependence. PMID- 12625294 TI - New female sterilization device. PMID- 12625295 TI - New guidelines focus on fish oil, omega-3 fatty acids. PMID- 12625296 TI - Cholesterol-lowering drug improves survival after heart transplant. PMID- 12625297 TI - Mild aerobic exercise no protection from osteoporosis. PMID- 12625298 TI - New FDA Commissioner sees challenges, opportunities. PMID- 12625299 TI - Turning up the heat on acrylamide. PMID- 12625300 TI - Drug research and children. PMID- 12625301 TI - FDA issues warning on decorative contact lenses. PMID- 12625302 TI - West Nile virus: reducing the risk. PMID- 12625303 TI - The lowdown on depression. PMID- 12625304 TI - MSG. A common flavor enhancer. PMID- 12625305 TI - Use caution with pain relievers. PMID- 12625306 TI - Wisconsin companies guilty of 'cheesy' operations. PMID- 12625307 TI - Pediatric drug research and mental illness. PMID- 12625308 TI - Community cooperatives combat sexual assault and domestic violence. AB - The effectiveness of the SANE program is borne out by the following testimonies: "The emotional support required by these victims is best rendered by a SANE. This frees the ED nurse to care for other patients, while sexual assault victims receive a high level of care," says Nancy Donel, RN manager at St. Thomas Hospital ED. "The DOVE program benefits not only the emergency physician, but the EMS system as well. It gives us a resource and a specifically identified program with well-trained, qualified providers. Through their training and knowledge, SANEs not only help victims, but also increase the number of legal convictions that take assailants off the streets. This improves the health and safety of the communities in which we live and serve," says Michael Mackan, MD, of the Summa Health System. PMID- 12625309 TI - Ultrastructural characteristics of the lung of Melanophryniscus stelzneri stelzneri (Weyenberg, 1875) (Anura, Bufonidae). AB - The lung of the toad, Melanophryniscus stelzneri stelzneri was studied using scanning and transmission electron microscopy. In M.s.stelzneri the parenchyma forms a polygonal network arrangement, therefore the parenchyma is edicular. These spaces are delimited by the interconnection of third order septa which are covered by respiratory epithelium. Small patches of ciliated epithelium without goblet cells appear irregularly distributed on the septa. The respiratory epithelium consists of one type of pneumocyte, which shows characteristics of both type I and type II alveolar cells of higher vertebrates. The pneumocytes are irregular in shape and possess attenuated cytoplasmic processes, which spread around the capillaries to form the outer layer of the air-blood barrier. These cells contain different types of cytoplasmic bodies: electron dense bodies, multivesicular bodies and lamellar bodies. Dense bodies are probably the precursors of lamellar bodies and the multivesicular bodies are incorporated into the latter. Neuroepithelial bodies appear randomly distributed over the septa. These bodies are separated from the lumen of the lung by thin cytoplasmic processes of neighbouring pneumocytes. The air-blood barrier consists of three layers: epithelium, interstitial space and endothelium. The relatively simple pulmonary structure of M.s.stelzneri is due to a lower degree of partitioning of the pulmonary lumen in comparison to the lung of other bufonid anurans, could be correlated with a well developed cutaneous and buccopharingeal respiration. The testing of this hypothesis awaits further studies. PMID- 12625311 TI - [Drugs with Iberis amara as a modern phytotherapy to treat chronic functional disorders of the gastrointestinal tract]. PMID- 12625310 TI - Prevention of propofol-induced injection pain by remifentanil: a placebo controlled comparison with lidocaine. AB - In a randomised, double-blind study we compared the efficacy of continuous remifentanil infusion (0.25 microg x kg(-1) x min(-1) with 40 mg lidocaine and placebo in the prevention of injection pain due to intravenous propofol administration (1.5-2 mg x kg(-1)) in 155 patients scheduled for elective surgery. Pain severity was evaluated using a four-point scale. The incidence of injection pain was 62% in the placebo group and could be reduced significantly by using remifentanil (30%; p < 0.0015) or lidocaine (33%; p < 0.005). Analysis of the pain scores showed a significant difference between remifentanil and placebo (p < 0.00005) as well as between lidocaine and placebo (p < 0.0002). There was no significant difference between remifentanil and lidocaine. Remifentanil provided effective pain relief, comparable with lidocaine, and is an alternative as part of an intravenous anaesthesia regimen to using another concomitant drug. PMID- 12625312 TI - Stanley F. Schneider (1922-2002). PMID- 12625313 TI - Studies shed light on ambulatory surgery risks. PMID- 12625314 TI - Teaching breast self-examination in developing countries does not reduce breast cancer deaths. PMID- 12625316 TI - Smoking quit lines work in clinical trials and actual practice. PMID- 12625315 TI - In re Marriage of Buzzanca: charting a new destiny. PMID- 12625317 TI - Blood, sweat, and tears: toward a new paradigm for protecting donor privacy. PMID- 12625318 TI - Poverty is a major factor in African American breast cancer deaths. PMID- 12625319 TI - Protecting health information privacy: the case for self-regulation of electronically held medical records. PMID- 12625320 TI - Physician-assisted suicide: the Supreme Court's wary rejection. PMID- 12625321 TI - Neade v. Portes: a physician's fiduciary duty to disclose financial incentives. PMID- 12625323 TI - Fertility control in wildlife. Proceedings of the Fifth International Symposium on Fertility Control in Wildlife. Skukuza, The Kruger National Park, South Africa. August 19-22, 2001. PMID- 12625322 TI - Oregon's guidelines for physician-assisted suicide: a legal and ethical analysis. AB - Oregon's Death with Dignity Act was first passed by a ballot initiative in 1994, but numerous judicial challenges delayed implementation of the Act. In November of 1997, following the United States Supreme Court decisions in Vacco v. Quill and Washington v. Glucksberg, which left the states' power to regulate physician assisted suicide undisturbed, the Oregon voters upheld their law. Oregon remains the only state in the nation to authorize physician-assisted suicide. The Task Force to Improve the Care of Terminally Ill Oregonians published a Guidebook for health care providers on the Oregon Act, and the New England Journal of Medicine recently issued a special report on the first year's experience under the Act. This paper analyzes the legal context of the Oregon Death with Dignity Act, discusses the efficacy of the tenets in the Guidebook, and explores ethical issues underlying the guidelines, particularly those pertaining to the meaning of a patient's request for assisted suicide and processes supporting informed consent. PMID- 12625324 TI - Unilateral lichen planus located on the chest showing a patchy and linear distribution. PMID- 12625325 TI - Onychotillomania. PMID- 12625326 TI - Proceedings of the Annual Conference of the International Embryo Transfer Society. Auckland, New Zealand. January 11-15, 2003. PMID- 12625329 TI - The old faith and the new science: the Nuremberg Code and human experimentation ethics in Britain, 1946-73. AB - This article explores the impact of the Nuremberg Code on post-Second World War research ethics in Britain. Against the background of the Nuremberg Medical Trial, the Code received international endorsement, but how much did its ethical percepts influence actual research? This paper shows that, despite British involvement in the formulation of the Code, the experience of war-time and changing career structures were more influential in shaping the approach of investigators to their subjects. Where medical debates ensued, primarily over controversial research practices at the British Postgraduate Medical School, Hammersmith Hospital, they were set in the context of a much older division between 'bedside' and 'scientific' medicine. But whatever differences there may have been between those physicians who advocated research and those who questioned its use and ethical basis, most clung to the paternalist assumption that it was the doctor's place to decide what was best for his patients. Faced with rising public and medical criticism of contemporary research practices, the medical elite of the 1960s and 1970s safeguarded the reputation of the profession and medical control of research by negotiating new voluntary codes. In a similar move, their predecessors had helped to negotiate the Nuremberg Code in anticipation of public criticism of experimentation arising from the Nuremberg Medical Trial. PMID- 12625328 TI - Understanding the origin of wound pain during dressing change. PMID- 12625327 TI - Cutaneous meningioma. PMID- 12625330 TI - The effects of input rate and synchrony on a coincidence detector: analytical solution. AB - We derive analytically the solution for the output rate of the ideal coincidence detector. The solution is for an arbitrary number of input spike trains with identical binomial count distributions (which includes Poisson statistics as a special case) and identical arbitrary pairwise cross-correlations, from zero correlation (independent processes) to complete correlation (identical processes). PMID- 12625331 TI - A theoretical model of axon guidance by the Robo code. AB - After crossing the midline, different populations of commissural axons in Drosophila target specific longitudinal pathways at different distances from the midline. It has recently been shown that this choice of lateral position is governed by the particular combination of Robo receptors expressed by these axons, presumably in response to a gradient of Slit released by the midline. Here we propose a simple theoretical model of this combinatorial coding scheme. The principal results of the model are that purely quantitative rather than qualitative differences between the different Robo receptors are sufficient to account for the effects observed following removal or ectopic expression of specific Robo receptors, and that the steepness of the Slit gradient in vivo must exceed a certain minimum for the results observed experimentally to be consistent. PMID- 12625332 TI - Verification of the selectivity of (+)N-3-benzylnirvanol as a CYP2C19 inhibitor. PMID- 12625333 TI - Reader consensus. PMID- 12625334 TI - Arising from the dead: challenges of posthumous procreation. AB - The medical capabilities derived from modern reproductive technology, such as in vitro fertilization and cryopreservation, have enabled physicians and scientists to intervene in the procreative process in innumerable ways. However, this intervention in the natural reproductive process raises both moral and legal concerns. In this Article, Professor Schiff explores some of the conflicts that may result when an individual or couple elects to cryopreserve gametes or embryos and subsequently, one or both of the contributors dies, or when gametes are harvested from a dead body. This Article will specifically address the moral and legal responses to circumstances where the decedent has either clearly expressed opposition to posthumous use of the reproductive material or else the decedent's intent regarding posthumous use of the material is ambiguous. By discussing philosophical and moral positions relating to personhood and the body and analyzing legal issues such as reproductive choice and organ donation, Professor Schiff creates the necessary format to examine and recommend the proper legal treatment of this controversial aspect of posthumous procreation. PMID- 12625336 TI - Is it less cruel if the animals 'want' to do it? PMID- 12625335 TI - Head to head: people with inherited diseases are ready to challenge pro-lifers over the future of medical research. PMID- 12625337 TI - The people vs patents. PMID- 12625338 TI - The scandal of IVF league tables. PMID- 12625339 TI - Your money or your life. PMID- 12625340 TI - Ethical dilemmas in testing for late onset conditions: reactions to testing and perceived impact on other family members. AB - This paper examines some of the ethical dilemmas that arise when testing for the late onset, untreatable condition of Huntington disease (HD) specifically now that technology allows us to test younger generations of the family for the mutant gene. Drawing on interview data from families with Huntington disease, the reactions to testing and perceived impact on other family members are considered. These are discussed in the light of the possible lowering of the age of test applicants. This potentially raises problems for a younger generation as they are confronted with reproductive decisions that the older generation did not have to face. It also means that individuals have the prospect of living with knowledge of a future illness for much longer. PMID- 12625342 TI - Small deep brain infarcts. PMID- 12625343 TI - Transnational pharmaceutical corporations and neo-liberal business ethics in India. AB - The author critiques the expedient application of market valuation principles by the transnational corporations and other large firms in the Indian pharmaceutical industry on a number of issues like patents, pricing, irrational drugs, clinical trials, etc. He contends that ethics in business is chiseled and etched within the confines of particular social structures of accumulation. An ascendant neo liberal social structure of accumulation has basically shaped these firms' sharp opposition to the Indian Patents Act, 1970, government administered pricing, etc. The author contents that the practice of neo-liberal economics is strongly associated with a "one-dimensional" ethics that privileges market valuation principles over all others. This seems to inevitably generate a social counter movement that struggles for social protections. He critiques neo-liberal business practices from a perspective that derives from the work of the economic anthropologist Karl Polanyi. Before the present phase of liberalization in India, markets were "managed", but without a "welfare state" in place. Moving toward deregulation of the markets without a welfare state in place is unethical. Keeping the debilities of the institutional framework of public policy in mind, the author adopts a Polanyian perspective that places its trust and hope in the growing social legitimacy of the counter-movement in opposition to both neo liberal business practices and the degenerate behavior of state agencies. PMID- 12625344 TI - Entrepreneurial "mining" of the dying: viatical transactions, tax strategies and mind games. AB - Conceptually, entrepreneurship is seen as the engine that drives a robust economy, promotes a favorable quality of life, and assures the availability of the attributes needed for meaningful living. However, like many popular concepts in this world, its limitations are normally not well acknowledged. A grouping of entrepreneurial ventures which as recently come into existence deals with the personal fiscal issues associated with the end-of-life phase of the human cycle. While generally praised as humanitarian services for society, that are assuredly within legal bounds, these activities have potentially significant negative implications. When examined from an ethical perspective, some questionable practices became apparent. Three such undertakings in particular have to do with the terminally ill and are examined in this paper. The first, viatical transactions, is defined as the transfer of a life insurance policy's ownership to someone who does not have an insurable interest in the life of the insured individual. The second, creative tax shelters for wealthy people involved in estate planning, involves hiring unrelated parties, particularly those who are young and in failing health, to do the dying for the principal. The third, mind games, involves curtailing the use of medical resources for the dying. The justification for this is rationing scarce medical resources. PMID- 12625345 TI - Occupational therapy for stroke patients: when, where, and how? PMID- 12625346 TI - [Dysphagia with vomiting. Highly differentiated liposarcoma of the larynx]. PMID- 12625347 TI - Advertising for clinical research. PMID- 12625348 TI - Practicing safer research: using the law to protect the confidentiality of sensitive research data. PMID- 12625349 TI - The ethics of clinical research in developing countries. PMID- 12625350 TI - Confidentiality of health care provider quality of care information. PMID- 12625351 TI - Vasculocentricity versus cerebrocentricity: what stroke-related baroreceptor reflex sensitivity changes might be telling us. PMID- 12625352 TI - The misuse of statistics: concepts, tools, and a research agenda. AB - This paper presents concerns regarding misuse of statistics in scientific work, especially in biomedical research. The paper discusses what is meant by "misuse." It appears that misuse arises from various sources: degrees of competence in statistical theory and methods, honest error in the application of methods, egregious negligence, and deliberate deception (misconduct.) The incidence of error is partly due to a perceived need to meet artificial statistical criteria for acceptance of research reports for publication by journals. There has been no systematic research into the prevalence of misuse or its breakdown by type. Nonetheless, there are ways to encourage, or even to enforce, good statistical practice. These can be greatly supported by use of available statistical ethics documents. This article suggests lines of further research that could define the problem more explicitly and that might lead to additional corrective measures. PMID- 12625354 TI - ICN leadership programme will reach more nurses. PMID- 12625353 TI - Informed consent and investigational new drug abuses in the U.S. military. AB - OBJECTIVE: The focal point of this investigation was to research the ethical issues surrounding the military's requests for informed consent waivers when using investigational drugs, and the recent debate surrounding the anthrax vaccine as an investigational new drug (IND). DESIGN: The military's management of the informed consent process was examined using documents obtained through the Freedom of Information Act, Institutional Review Board (IRB) minutes, legal pleadings, and protocols for specific investigational drugs. RESULTS: In December 1990, prior to Operation Desert Storm, the Federal Drug Administration (FDA) granted the Department of Defense (DoD) an unprecedented waiver to the federally mandated informed-consent requirement for the use of investigational drugs. However, the waiver approval was conditional, and the FDA insisted on several safeguards. Partially in response to the subsequent Gulf War Syndrome debate, the FDA recently evaluated the military's use of investigational drugs during the Gulf War. The FDA cited the military for significant deviations from the originally approved protocols. Most notably, the military was found to be abusing the IRB process by convening a second IRB when the first IRB concluded that waiving informed consent was unethical. In addition, there was a gross lack of documentation and no monitoring of adverse reactions. The DoD's plan to use the current anthrax vaccine on all 2.4 million troops against inhalation anthrax has kindled an additional investigational drug controversy. The safety and efficacy of the use of the anthrax vaccine as a prophylactic against inhalation anthrax have been questioned by both military and medical organizations. There have never been any published studies of human efficacy or long-term effects for the anthrax vaccine. In addition, the military is not using the vaccine for its intended purpose, and it is also not adhering to prescribed dosing schedules. There is clear evidence to support the claim that, in fact, the military's use of the anthrax vaccine should be considered unethical. CONCLUSIONS: I argue that in medical situations, the military is obligated to treat its troops as autonomous persons entitled to basic rights and protections. The DoD is currently using an approved drug, the anthrax vaccine, for an unapproved purpose and in an unapproved manner. In doing so, the DoD is not only violating the FDA's regulations against such practices, it is also violating an executive order which only allows the president to authorize the use of INDs on service members without their consent. PMID- 12625355 TI - Rural Africa benefits from ICN mobile libraries. International Council of Nurses. PMID- 12625356 TI - "Thank God for the lawyers": some thoughts on the (mis)regulation of scientific misconduct. PMID- 12625357 TI - [Physiopathology and management in children--indication for tonsillectomy]. PMID- 12625358 TI - A tale of two experts: thalidomide and political engagement in the United States and West Germany. AB - The physicians, Widukind Lenz and Frances Kelsey, played crucial roles in the thalidomide drama of the early 1960s. Widespread use of the drug in West Germany was only halted when the paediatrician, Lenz, publicized its association with the birth of nearly 4,000 children exhibiting abnormal limb growth. Few cases were reported in the United States because Kelsey, a medical officer at the US Food and Drug Administration, repeatedly delayed thalidomide's marketing approval. Experts in both countries were expected to demonstrate publicly the professional 'objectivity' of medicine and the institutional 'disinterestedness' of regulatory bodies. These norms were invoked both by industry representatives seeking to undermine the two experts and by critics desiring stronger regulatory controls. Comparing Lenz and Kelsey demonstrates how institutional structures shape an expert's social and scientific roles. While the United States provided important protection from external pressure for Kelsey through her regulatory position at the FDA, Lenz was open to sharp criticism, especially when giving expert testimony during a lengthy court trial. The degree of exposure to politically motivated attacks differed for these two experts; they nevertheless faced similar threats to their professional credibility and personal integrity when they publicized links between thalidomide and birth defects. PMID- 12625359 TI - Beverages: bottled water. Direct final rule. AB - The Food and Drug Administration (FDA) is amending its bottled water quality standard regulations by establishing an allowable level for the contaminant uranium. As a consequence, bottled water manufacturers are required to monitor their finished bottled water products for uranium at least once each year under the current good manufacturing practice (CGMP) regulations for bottled water. Bottled water manufacturers are also required to monitor their source water for uranium as often as necessary, but at least once every 4 years unless they meet the criteria for the source water monitoring exemptions under the CGMP regulations. FDA will retain the existing allowable levels for combined radium 226/-228, gross alpha particle radioactivity, and beta particle and photon radioactivity. This direct final rule will ensure that the minimum quality of bottled water, as affected by uranium, combined radium-226/-228, gross alpha particle radioactivity, and beta particle and photon radioactivity, remains comparable with the quality of public drinking water that meets the Environmental Protection Agency's (EPA's) standards. FDA is issuing a direct final rule for this action because the agency expects that there will be no significant adverse comment on this rule. Elsewhere in this issue of the Federal Register, FDA is publishing a companion proposed, rule under the agency's usual procedure for notice-and-comment rulemaking, to provide a procedural framework to finalize the rule in the event the agency receives any significant adverse comments and withdraws this direct final rule. The companion proposed rule and direct final rule are substantively identical. PMID- 12625360 TI - Experimental rage: the development of medical ethics and the genesis of scientific facts. Ludwik Fleck: an answer to the crisis of modern medicine in interwar Germany? AB - In 1930, the large-scale introduction of the BCG vaccination in the city of Lubeck in northern Germany led to a major scandal that focused public attention on medical experimentation with human beings as well as reviving criticism of the medical profession that had been voiced before. The trial following the catastrophe raised the first clearly identifiable public discussions on medical ethics in Europe, and led to the establishment of the first regulations for medical research on human beings in the western hemisphere; the German 'Richtlinien' of 1931. In 1935, Ludwik Fleck (1896-1961) published a now classic monograph entitled Genesis and Development of a Scientific Fact. The central hypothesis of this article is that, when Fleck published his book four years after the Lubeck trial, he was proposing answers to questions raised, at least partially, by the Lubeck case, although he never explicitly mentions it. Most interestingly, Fleck proposed a different approach to the fundamental dilemma of modern experimental medicine, the potential opposition between an individual's well-being, and the production and application of scientific knowledge in medicine. Where the standard answer to these questions has, since the 1930s, become moral reasoning and ethical regulation, known today as bioethics, Fleck portrays a different approach that could be characterized as the attempt to foster a deeper and more democratic understanding of science through an examination of its intimate functioning. PMID- 12625361 TI - Health claims and other health-related statements in the labeling and advertising of alcohol beverages (99R-199P). Final rule, Treasury decision. AB - TTB is amending the regulations to prohibit the appearance on labels or in advertisements of any health-related statement, including a specific health claim, that is untrue in any particular or tends to create a misleading impression. A specific health claim on a label or in an advertisement is considered misleading unless the claim is truthful and adequately substantiated by scientific evidence; properly detailed and qualified with respect to the categories of individuals to whom the claim applies; adequately discloses the health risks associated with both moderate and heavier levels of alcohol consumption; and outlines the categories of individuals for whom any levels of alcohol consumption may cause health risks. In addition, TTB will consult with the Food and Drug Administration (FDA), as needed, on the use of specific health claims on labels. If FDA determines that a specific health claim is a drug claim that is not in compliance with the requirements of the Federal Food, Drug, and Cosmetic Act, TTB will not approve the use of such statement on a label. Health related statements that are not specific health claims or health-related directional statements will be evaluated on a case-by-case basis to determine if they tend to mislead consumers. The final rule provides that health-related directional statements (statements that direct or refer consumers to a third party or other source for information regarding the effects on health of alcohol consumption) will be presumed misleading unless those statements include a brief disclaimer advising consumers that the statement should not encourage consumption of alcohol for health reasons, or some other appropriate disclaimer to avoid misleading consumers. TTB believes that the final regulations will ensure that labels and advertisements do not contain statements or claims that would tend to mislead the consumer about the significant health consequences of alcohol consumption. PMID- 12625362 TI - [Proof of efficacy as basis for assessment of diagnostic and therapeutic methods in medical expert testimony]. PMID- 12625363 TI - [A 64-year-old man with pre- and pan-systolic murmurs after acute myocardial infarction]. PMID- 12625364 TI - Protease-activated receptors: a means of converting extracellular proteolysis into intracellular signals. AB - Protease-activated receptors (PARs) mediate cellular responses to a variety of extracellular proteases. The four known PARs constitute a subgroup of the family of seven-transmembrane domain G protein-coupled receptors and activate intracellular signalling pathways typical for this family of receptors. Activation of PARs involves proteolytic cleavage of the extracellular domain, resulting in formation of a new N terminus, which acts as a tethered ligand. PAR 1, -3, and -4 are relatively selective for activation by thrombin whereas PAR-2 is activated by a variety of proteases, including trypsin and tryptase. Recent studies in mice genetically incapable of expressing specific PARs have defined roles for PAR-1 in vascular development, and for PAR-3 and -4 in platelet activation, which plays a fundamental role in blood coagulation. PAR-1 has also been implicated in a variety of other biological processes including inflammation, and brain and muscle development. Responses mediated by PAR-2 include contraction of intestinal smooth muscle, epithelium-dependent smooth muscle relaxation in the airways and vasculature, and potentiation of inflammatory responses. The area of PAR research is rapidly expanding our understanding of how cells communicate and control biological functions, in turn increasing our knowledge of disease processes and providing potential targets for therapeutic intervention. PMID- 12625365 TI - The complex nature of protein phosphatases. AB - Protein phosphatases are integrally associated with the regulation of cellular signaling. The mechanisms underlying the specific regulatory roles are likely to be unique to each cell system. Nevertheless, analysis of phosphatase regulation in a number of systems has identified phosphatase targeting through association with a wide range of binding partners to be a fundamental mechanism of regulation. Using protein phosphatase 2A (PP2A) as an example, this snapshot summarizes these fundamental mechanisms of protein phosphatase regulation. PMID- 12625366 TI - Do hemoglobin and hemocyanin impair schistosoma killing by no? AB - Nitric oxide (NO) possesses antiparasitic effects on both Protozoa and Metazoa in definitive and intermediate hosts as well as in vectors. Here, we postulate that hemoglobin and hemocyanin may impair Schistosoma killing by NO in the definitive and intermediate hosts. Interestingly, hemoglobin, myoglobin, and neuroglobin may protect Plasmodium and Trypanosoma from the antiparasitic effects of NO. PMID- 12625367 TI - Nutrient-gene interactions in mitochondrial function: vitamin A needs are increased in BHE/Cdb rats. AB - The BHE/Cdb rat has a maternally inherited mutation in the ATPase 6 mitochondrial gene that associates with impaired oxidative phosphorylation (OXPHOS) and glucose intolerance. A longevity study revealed that feeding an egg-rich (vitamin A-rich) diet delayed the onset of impaired glucose tolerance. Two experiments were conducted to test the hypothesis that BHE/Cdb rats require more dietary vitamin A than normal rats. Experiment 1 was a dose-response study examining OXPHOS in BHE/Cdb rats fed one of six levels of vitamin A. In experiment 2 BHE/Cdb and Sprague-Dawley rats were used. The rats were depleted of retinol stores, then repleted with 4 or 12 IU vitamin A/g diet. Vitamin A status was assessed in depleted, never depleted, and depleted/repleted rats. OXPHOS was optimized at 4 IU/g diet for the Sprague-Dawley rats and 12 IU/g diet for the BHE/Cdb rats. These results suggested that the criteria for vitamin intake adequacy in the BHE/Cdb rats is the optimization of mitochondrial OXPHOS. Using this criteria, we conclude that diabetes-prone BHE/Cdb rats require more dietary vitamin A than normal rats. PMID- 12625368 TI - Nutrient-gene interactions: dietary vitamin A and mitochondrial gene expression. AB - The BHE/Cdb rat is a model for mitochondrial diabetes due to a mutation in the ATPase 6 gene. These rats require more dietary vitamin A to optimize mitochondrial function than do normal Sprague-Dawley rats. To determine a possible mechanism for this effect, cultured hepatocytes and hepatic tissues were studied. ATPase 6 (F0ATPase subunit a), retinoic acid receptors (RARs), and mitochondrial transcription factor A (mtTFA) gene products were determined using Western blot analysis. Northern analysis was used to determine ATPase 6, ATPase 6,8, and ND1 mRNA. Mitochondrial density was determined using confocal microscopy. Dose response studies using primary hepatocyte cultures showed that both ATPase 6 gene product and mRNA were optimized with additions of 10(-9) M retinoic acid. Retinoic acid receptors were found in the mitochondrial compartment. MtTFA levels were increased by vitamin A. Mitochondrial density was greater in the BHE/Cdb tissue than in Sprague-Dawley tissue. These results show that vitamin A affects mitochondrial function via an effect on both nuclear and mitochondrial encoded genes. PMID- 12625369 TI - Suppression of gastric mucosal inflammatory responses to Helicobacter pylori lipopolysaccharide by peroxisome proliferator-activated receptor gamma activation. AB - Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is a nuclear transcription factor that regulates the expression of genes associated with inflammation. We applied the animal model of H. pylori lipopolysaccharide(LPS) induced gastritis to assess the effect of a specific PPAR-gamma ligand, ciglitazone, on the apoptotic processes and the mucosal activity of inducible nitric oxide synthase (NOS-2), and the expression of COX-1 and -2 cyclooxygenases. In the absence of ciglitazone, the LPS-elicited mucosal inflammatory responses were accompanied by a massive epithelial cell apoptosis, upregulation of NOS-2 and COX-2 protein expression, and a marked increase in the mucosal PGE2 generation and NOS-2 activity. The expression of COX-1 protein, however, remained unchanged. Administration of ciglitazone led to dose-dependent reduction (up to 48%) in the severity of mucosal inflammatory involvement elicited by the LPS and this effect of the agent was reflected in a 72.5% reduction in apoptosis, a 58.7% decline in the mucosal PGE2 generation and a 75.6% drop in NOS-2 activity, and produced a marked decrease inCOX-2 and NOS-2 protein expression. Our findings demonstrate that PPAR-gamma activation suppresses gastric mucosal inflammatory responses to H. pylori LPS, and suggest that pharmacological manipulation of PPAR-gamma activation may provide therapeutic benefits in the resolution of inflammation associated with H. pylori infection. PMID- 12625370 TI - E2F1 activates the human p53 promoter and overcomes the repressive effect of hepatitis B viral X protein (Hbx) on the p53 promoter. AB - The functional effect of the interaction of E2F1 and hepatitis B virus X protein (HBx) on the promoter of human p53 gene was studied using chloramphenicol acetyl transferase (CAT) assay. E2F1 activated the p53 promoter through E2F1 binding site. As previously reported, HBx repressed the p53 promoter through E-box. When E2F1 was cotransfected with HBx, E2F1 overcame the repressive effect of HBx on the p53 promoter through the E2F1 site. However, in the thymidine kinase (tk) heterologous promoter system with the E2F1 binding sites, cotransfection of E2F1 and HBx showed a strong synergistic activation. An in vitro interaction assay showed that E2F1 and HBx physically bind with each other. Analyses of the interaction domain with the GAL4 fusion protein showed that the pRb-binding domain of E2F1 was necessary for the functional interaction of these two proteins. Taken together, these results imply the functional inhibitory action of E2F1 on the HBV life cycle and HBV-mediated hepatocellular carcinogenesis (HCC). Therefore, the normal or enhanced function of E2F1 gene would be important in controlling the HBx function in HCC. PMID- 12625371 TI - Would mammalian fructose metabolism be more sensible if fructose was converted into fructose-6-phosphate rather than the -1-phosphate? PMID- 12625372 TI - How I became a biochemist. PMID- 12625373 TI - Monitoring dyslexics' intelligence and attainments: a follow-up study. AB - Intelligence (Wechsler Intelligence Scale for Children R and III, 1992) and written language attainment (BAS Word Reading, Neale Analysis of Reading, Vernon Graded Word Spelling) data for around 250 children attending a specialist school for dyslexics are presented. The Wechsler scales data show some evidence for 'ACID' and 'SCAD' profile effects on the subtests, with specifically weak Index scores on Freedom from Distractibility and Processing Speed. The relationship between intelligence and reading development is also examined, with evidence for significant correlations between intelligence and written language and a longitudinal study showing that there is no 'Matthew' or drop-off effect in intelligence. The attainments tests demonstrate that the widening gap between a dyslexic's chronological age and his/her attainments can be closed, and how attainments may be monitored within the context of 'growth curves'. The results are discussed in relation to recent reports (e.g. B.P.S. on Dyslexia, Literacy and Psychological Assessment) on the relationship between intelligence and attainments and it is concluded that this report could be seriously misleading for practising educational psychologists. PMID- 12625374 TI - Dyslexia and music: measuring musical timing skills. AB - Over the last few decades, a growing amount of research has suggested that dyslexics have particular difficulties with skills involving accurate or rapid timing, including musical timing skills. It has been hypothesised that music training may be able to remediate such timing difficulties, and have a positive effect on fundamental perceptual skills that are important in the development of language and literacy skills (Overy, 2000). In order to explore this hypothesis further, the nature and extent of dyslexics' musical difficulties need to be examined in more detail. In the present study, a collection of musical aptitude tests (MATs) were designed specifically for dyslexic children, in order to distinguish between a variety of musical skills and sub-skills. 15 dyslexic children (age 7-11, mean age 9.0) and 11 control children (age 7-10, mean age 8.9) were tested on the MATs, and their scores were compared. Results showed that the dyslexic group scored higher than the control group on 3 tests of pitch skills (possibly attributable to slightly greater musical experience), but lower than the control group on 7 out of 9 tests of timing skills. Particular difficulties were noted on one of the tests involving rapid temporal processing, in which a subgroup of 5 of the dyslexic children (33%) (mean age 8.4) was found to account for all the significant error. Also, an interesting correlation was found between spelling ability and the skill of tapping out the rhythm of a song, which both involve the skill of syllable segmentation. These results support suggestions that timing is a difficulty area for dyslexic children, and suggest that rhythm skills and rapid skills may need particular attention in any form of musical training with dyslexics. PMID- 12625375 TI - Age-related improvements in auditory temporal resolution in reading-impaired children. AB - Individuals with developmental dyslexia show impairments in processing that require precise timing of sensory events. Here, we show that in a test of auditory temporal acuity (a gap-detection task) children ages 6-9 years with dyslexia exhibited a significant deficit relative to age-matched controls. In contrast, this deficit was not observed in groups of older reading-impaired individuals (ages 10-11 years; 12-13 years) or in adults (ages 23-25 years). It appears, therefore, that early temporal resolution deficits in those with reading impairments may significantly ameliorate over time. However, the occurrence of an early deficit in temporal acuity may be antecedent to other language-related perceptual problems (particularly those related to phonological processing) that persist after the primary deficit has resolved. This result suggests that if remedial interventions targeted at temporal resolution deficits are to be effective, the early detection of the deficit and early application of the remedial programme is especially critical. PMID- 12625376 TI - Evaluation of an exercise-based treatment for children with reading difficulties. AB - An evaluation is reported of an exercise-based approach to remediation of dyslexia-related disorders. Pupils in three years of a Warwickshire junior school were screened for risk of literacy difficulty using the Dyslexia Screening Test (DST). The 35 children scoring 0.4 or over on the DST were divided randomly into two groups matched for age and DST score. One quarter of the participants had an existing diagnosis of dyslexia, dyspraxia or ADHD. Both groups received the same treatment at school but the intervention group used the DDAT exercise programme daily at home. Performance on the DST and specialist cerebellar/vestibular and eye movement tests were assessed initially and after six months. Cerebellar/vestibular signs were substantially alleviated following the exercise treatment whereas there were no significant changes for the control group. Even after allowing for the passage of time, there were significant improvements for the intervention group in postural stability, dexterity, phonological skill, and (one-tailed) for naming fluency and semantic fluency. Reading fluency showed a highly significant improvement for the intervention group, and nonsense passage reading was also improved significantly. Significantly greater improvements for the intervention group than the control group occurred for dexterity, reading, verbal fluency and semantic fluency. Substantial and significant improvements (compared with those in the previous year) also occurred for the exercise group on national standardized tests of reading, writing and comprehension. It is concluded that, in addition to its direct effects on balance, dexterity and eye movement control, the benefits of the DDAT exercise treatment transferred significantly to cognitive skills underlying literacy, to the reading process, and to standardized national literacy attainment tests. PMID- 12625377 TI - The prevalence and economic importance of bovine fasciolosis in Kenya--an analysis of abattoir data. AB - A retrospective study covering a period of 10 years (1990-1999) was carried out using post mortem meat inspection records at the Veterinary Department Headquarters at Kabete to determine the prevalence and economic importance of bovine fasciolosis in Kenya. Meat inspection records from abattoirs in 38 districts distributed over seven out of the eight provinces of Kenya were examined. Prevalence of fasciolosis was calculated as the number of cattle found to be infected with Fasciola, expressed as a percentage of the total number of cattle slaughtered. Using the average weight and market price of a bovine liver, the monetary loss occasioned by condemnation of Fasciola infected livers was calculated. A survey was also carried out at Dagoretti slaughterhouse complex in Nairobi to determine the relative occurrence of F. gigantica and F. hepatica in slaughtered cattle. Cattle slaughtered at Dagoretti slaughterhouse originate from all parts of the country. A total of 5,421,188 cattle were slaughtered in the seven provinces of Kenya during the 10-year period and 427,931 (8%) of these cattle were infected with Fasciola. The region with the highest prevalence of fasciolosis was Western Province (16%) followed, in descending order, by Eastem Province (11%), Nyanza Province (9%), Rift Valley Province (8%), Central Province (6%), Nairobi Province (4%) and Coast Province (3.5%). The total economic loss incurred by the country during the 10-year period as a result of condemnation of the infected livers was approximately US$2.6 million. The total annual economic losses during this period ranged from approximately US$0.2-0.3 million. The highest total economic losses for the 10-year period were recorded in Western Province (US$0.8 million) and Central Province (US$0.7 million). A total of 1584 cattle originating from five provinces of Kenya were slaughtered at Dagoretti slaughterhouse over a period of two months of which 147 (9.3%) were infected with liver flukes. All the liver flukes obtained from the infected livers were identified as F. gigantica. It is concluded that fasciolosis is prevalent in cattle in all provinces of Kenya, that it causes great economic losses as a result of condemnation of infected livers, and that F. gigantica is the main species of liver flukes affecting cattle in Kenya. Local climatic factors, cattle trade, rustling and population numbers, and the presence of the snail intermediate hosts are probably the main factors influencing the incidence of the disease in the various regions of the country. PMID- 12625378 TI - Animal-level risk factors for Trypanosoma evansi infection in camels in eastern and central parts of Kenya. AB - Point prevalences and animal-level risk factors for Trypanosoma evansi infection were investigated in a cross-sectional study that involved 2227 camels from eastern and central parts of Kenya. The screening tests used were haematocrit centrifugation technique (HCT), mouse inoculation and latex agglutination (Suratex). All camels were screened with HCT, while 396 and 961 of them were, in addition, screened with mouse inoculation and Suratex tests, respectively. Parasitological and Suratex test results were used in parallel to determine the number of camels exposed to T. evansi infections. Statistical analyses were conducted using Statistical Analysis Systems. Parasitological and Suratex test results in parallel were dependent variables in multivariable logistic regression models that determined risk factors for T. evansi infection. Herd-level clustering was corrected with general estimation equations. The prevalences were 2.3% and 19.6%, using parasitological and Suratex tests, respectively, and 21.7% when both tests were used in parallel. There was a positive association between the screening tests (McNemar's test = 104.8, P = 0.001) although the strength of association was low (Kappa = 0.2; 95% CI: 0.1-0.3). Before accounting for herd level clustering, dry season (OR = 1.5; 95% CI: 1.0, 2.1) and nomadic pastoralism (OR = 1.8; 95% CI: 1.1, 3.2) were associated with increased odds of a camel being exposed to T. evansi infection compared to wet season and ranching, respectively. Following this correction, only nomadic pastoralism was significantly associated (OR = 3.1; 95% CI = 1.0, 14.4) with T. evansi infection compared to ranching. It is concluded that camels managed under nomadic pastoralism had higher risk of being exposed to T. evansi infections than camels from ranching systems of management. PMID- 12625379 TI - On-host ecology and off-host survival of the sheep scab mite Psoroptes ovis. AB - These studies were conducted to investigate the possible role of certain aspects of the on-host ecology and off-host survival of the sheep scab mite, Psoroptes ovis, in the dissemination of infestation. All developmental stages of the mite occurred in the fleece both proximally or distal to the skin of infested Merino and Dorper sheep. A larger proportion of mites was present in the fleece of Dorper sheep distal to the skin in the late afternoon and early morning than at other times during the day. Immature and adult mites readily transferred to tufts of wool or hair placed on infested sheep of both breeds. No mites could be found on wool or hair rubbed off onto tree trunks or branches or other structures in enclosures housing heavily infested sheep, nor could any mites be collected from the soil of these enclosures, whereas more than 80% of mites artificially seeded onto soil samples were recovered. The longest mean off-host survival times for larvae, nymphs, and male and ovigerous female mites were recorded at 10 degrees C, and were 9.25 days (RH = 90%), 15 days (RH = 33% and 75%), 10.5 days (RH = 75% and 90%) and 11.25 days (RH = 90%) respectively. Under natural climatic conditions ovigerous females in glass vials containing Merino wool survived for 17 days compared to 15 days for females in vials without wool; this difference was, however, not significant. The mean off-host pre-hatch period for eggs varied between 5.9 days (T = 25 degrees C and RH = 33%) and 22.1 days (T = 10 degrees C and RH = 75%), while the longest time individual eggs took to hatch at the latter temperature and RH was 31 days. PMID- 12625380 TI - Effects of the scab mite Psoroptes ovis on the haematology and live mass of Merino and Dorper sheep. AB - Five Merino and five Dorper sheep were artificially infested with the sheep scab mite Psoroptes ovis and the effect of infestation on their haematology, serum protein levels and live mass recorded for a period of 14 weeks. The reaction of the Merino sheep to infestation was more severe than that of the Dorper sheep. Haematological values fluctuated within the normal range during the assessment period. The mean haemoglobin concentration of the Merino sheep declined until antiparastic treatment was administered 10 weeks after infestation, after which it gradually increased. The lymphocyte counts of both breeds of sheep declined from 2 weeks to 10 weeks post-infestation, but increased after treatment, while the highest eosinophil counts were recorded in the Merino sheep at the height of the acute disease 8-10 weeks post-infestation. Serum albumin values for both breeds and serum globulin values for the Merino sheep were higher than normal during the entire 14-week observation period. A decrease in serum albumin and an increase in serum globulin concentration occurred at the height of infestation in both breeds. The mean live mass of a second group of five infested Merino sheep decreased by 6.4 kg over a 16-week period compared to a gain of 4.56 kg for five infested Dorper sheep. PMID- 12625381 TI - Apparent prevalence of dourine in the Khomas region of Namibia. AB - A 15-year record of the results of horse sera from the Khomas region of Namibia tested by the complement fixation test for dourine at the Central Veterinary Laboratory in Windhoek before clearing the respective animals for export and competitive sport were subjected to statistical analysis. The range of percentage positive, taken as the apparent prevalence of dourine for the region, during the period of study, was 0-29.09%; the average regional level of apparent prevalence was 8.33%. These figures were thought to be lower than the real situation due to some bias in the sampling criteria. For more accurate results, the more reliable enzyme-linked immunosorbent assay techniques are recommended for use in sero surveys for dourine in Khomas and other regions of Namibia to provide a basis for development of effective control strategies against the disease. PMID- 12625382 TI - Freezing of sheep faeces invalidates Haemonchus contortus faecal egg counts by the McMaster technique. AB - Faecal pellets from a sheep that was artificially infected with a monoculture of Haemonchus contortus were collected over a 2-h period in the morning. In the laboratory the faeces were thoroughly mixed by hand and 48 by 1 g aliquots of the pellets were sealed in plastic bags, from which the air had gently been expressed. The faecal worm egg count of the sheep was about 14,000 g(-1). Varying numbers of the bags were either processed for faecal worm egg counting (FEC) by the McMaster technique on day 0, or were stored at one of the following temperatures: about 4 degrees C, -10 degrees C or -170 degrees C before processing. The faecal aliquots that were frozen were thawed at room temperature after having been frozen for either 2 h or 7 days, and processing of aliquots maintained at 4 degrees C proceeded shortly after the samples had been removed from the refrigerator. A dramatic reduction in egg numbers was found in all the aliquots that were frozen at -170 degrees C before faecal worm egg counts were done, as well as in those frozen for 7 days at about -10 degrees C. Numerous empty, or partially empty, egg shells were observed when performing the counts in faeces that had been frozen. In contrast, there was no significant reduction in the numbers of eggs in aliquots maintained for 7 days in a refrigerator at +/- 4 degrees C before examination, when compared with others examined shortly after collection of the faeces. Since H. contortus eggs in faeces are damaged by freezing, some methods that can be used for short term preservation are outlined. It is concluded that all nematode egg counts from cryopreserved faeces (whether in a freezer at -10 degrees C or in liquid nitrogen) should possibly be regarded as being inaccurate, unless the contrary can be demonstrated for different worm genera. However, exceptions are expected for the more rugged ova, such as those of the ascarids and Trichuris spp. PMID- 12625383 TI - Review of current problems and shortcomings in the Tanzanian animal health information system with suggestions on improvement. AB - Livestock diseases have always been the focus of the Tanzanian Veterinary Authorities. However, they have become more important since the formation of the World Trade Organisation and subsequent implementation of the various multilateral agreements on trade. There is also a strong political desire to improve the animal health status as part of poverty alleviation strategies. As a result there is a need to develop better systems for investigating and reporting of animal diseases. In order to follow the OIE pathway and to obtain a disease free status, reliable evidence of freedom from particular diseases is becoming an issue of major interest. Assessment of the Tanzanian animal health information system revealed two major problems; firstly, the absence of disease information that accurately reflects the health status of the source population, and secondly, an inefficient information management system which is unable to provide useful information on the spatial component of animal health. A strategic approach is proposed that involves the collection of animal health information using active surveillance techniques and the introduction of a geographic information system. This approach should improve the management and reporting of animal health information. PMID- 12625384 TI - Water absorption in relation to fermentation in the colon of the ostrich (Struthio camelus). AB - The colon is a major site for fermentation and water absorption in the ostrich. Water absorption along the colon was evaluated and its relationship to osmolality, Na+ concentration, short chain fatty acid (SCFA) concentration and carbohydrate content of digesta analysed. Mean water content decreased from 5.30 +/- 0.99 to 2.51 +/- 0.13 mf/g dry mass in the first 5 m of the colon. Correspondingly, mean carbohydrate content fell from 529.85 +/- 46.61 to 434.99 +/- 29.89 mg/g dry mass. A significant correlation was shown between the decreases in mean carbohydrate and water content along the colon (r2 = 0.997, P < 0.05). Changes in mean osmolality (+/- 10 mOsm/kg) and SCFA concentration (+/- 7 mmol/l) were minimal in comparison to the change in Na+ concentration (-54 mmol/l). These findings reflect a close coupling between SCFA production and absorption on the one hand and water absorption on the other. PMID- 12625385 TI - Continuous exposure to an aversive mixture as a means of maintaining aversion to vermeerbos (Geigeria ornativa O. Hoffm.) in the presence of non-averted sheep. AB - Continuous exposure to an aversive mixture was investigated as a means of maintaining aversion to vermeerbos in sheep subjected to the social influence of non-averted sheep. The use of an aversive mixture was based on a hypothesis that continuous exposure to an acceptable aversive mixture (containing both the aversive substance and the identification factors of vermeerbos mixed with maize meal) would tempt sheep to consume small quantities of the aversive mixture each day and that this would keep them averted to vermeerbos, despite the social influence of non-averted sheep. Persistent aversion to a vermeerbos-maize meal mixture (1:99 by mass) by sheep continuously exposed to such an aversive mixture, after an initial aversion conditioning with lithium chloride (LiCl, 160 mg/kg BM), was demonstrated. Aversion in adjacent controls not exposed to the aversive mixture only lasted for some time. A similar result was obtained when sheep were challenged for intake of a pure stand of established vermeerbos. Three sheep continuously exposed to an aversive mixture after an initial aversion conditioning totally refused grazing the vermeerbos during a 42-day trial, despite the social influence of three non-averted control sheep grazing vermeerbos on an adjacent site. These results were confirmed by a second replication the following year. Joint grazing for an hour a day by averted and non-averted sheep during the last seven days of this replication also resulted in total avoidance of vermeerbos by the averted animals, despite continued intake of vermeerbos by the control sheep PMID- 12625387 TI - Early nerve repair after injury to the postganglionic plexus: an experimental study of sensory and motor neuronal survival in adult rats. AB - The optimal time for brachial plexus nerve repair is debatable. In this study we examined whether early re-establishment of neurotrophic support from the periphery might reduce neuronal loss. In 14 adult rats, the C7 spinal nerve was transsected. All sensory cells of the dorsal root ganglion and spinal motor neurons projecting into the C7 nerve were labelled retrogradely. The proximal and distal portions of the C7 nerve were then reanastomosed by either primary repair or by a vascularised or conventional ulnar nerve graft. At 16 weeks postoperatively, the nerve repair had significantly reduced the loss of both sensory and motor C7 neurons. Most striking was that a 30% motor neuronal loss in the control was almost eliminated by early nerve repair. In the grafted animals, half of the surviving neurons had regenerated through the graft, with no difference between vascularised and conventional nerve grafts. These results suggest that early surgical intervention may promote neuronal survival and regeneration after injuries to the brachial plexus. PMID- 12625386 TI - Amphistome infection of goats farmed under resource-poor conditions in South Africa. AB - From December 1998 to April 2000, a longitudinal study was conducted of the pooled trematode faecal egg counts of samples collected from goats of resource poor farmers at Rust de Winter, Gauteng Province, Impendle, KwaZulu-Natal Province, and Kraaipan, North-West Province, South Africa. The amphistome faecal egg counts followed a seasonal pattem, with an increase in the counts during the warmer months of the year (October to March). This is the first work concerning the seasonal cycling of amphistomes in ruminants in South Africa. PMID- 12625388 TI - Efficacy of topical nitroglycerin and transcutaneous electrical nerve stimulation on survival of random-pattern skin flaps in rats. AB - We compared the efficacy of topical nitroglycerin and transcutaneous electrical nerve stimulation (TENS) on the survival of random-pattern skin flaps in rats. Thirty Wistar albino rats were used and a dorsal, cranially-based random-pattern flap was raised. The rats were divided into three groups of 10 rats each. The first group had only the flap raised while the second and third groups were given topical nitroglycerin 5 mg or TENS for one hour a day for seven days. The amount of flap necrosis was measured on the seventh postoperative day. The mean area of necrosis in the flaps were 726.2, 544.2, and 150.0 mm2 in the control, nitroglycerin, and TENS groups, respectively. The mean percentage of flaps that necrosed were 51.9, 38.9, and 10.7 in the corresponding groups. The TENS group had significantly higher percentage area of flap surviving than the control (p < 0.0001) and nitroglycerin groups (p = 0.002). TENS, with its efficacy on survival and with negligible side-effects, could be a reliable treatment. Clinically, it can easily be used postoperatively when flaps become ischaemic, and will be tolerated well by patients. PMID- 12625389 TI - Effects of neurotrophic factors on chemokinesis of Schwann cells in culture. AB - Schwann cells are support cells in the peripheral nervous system and are responsible for migration, adhesion, production of the extracellular matrix, and myelination. Migration is considered to be essential for nerve regeneration after transsection. We have examined the chemokinetic effects of nerve growth factor (NGF), brain derived growth factor (BDNF), and neurotrophin-3 (NT-3) on Schwann cells in vitro using a chemotaxis chamber. The chemokinetic activity of Schwann cells was strongly accelerated by NGF, but was not influenced by BDNF. NT-3 at a concentration of 1 ng/ml had a stimulatory effect on chemokinesis. These data suggest that NGF is a chemoattractant for Schwann cells in vitro. Giving exogenous NGF might stimulate both migration of Schwann cells and the formation of Bungner's bands after peripheral nerve injuries in animal models. PMID- 12625390 TI - Periorbital reconstruction using variations of the upper eyelid flap. AB - Tissues that have characteristics identical or similar to periorbital soft tissue are preferred for reconstruction in the eye region. The upper eyelid is part of the periorbital area that provides an ideal reserve of tissue for this purpose, without producing a donor-site deformity. We used the upper eyelid flap with different pedicle designs in various parts of the periorbital region in 21 patients, and achieved favourable results. The flap was medially-based in five, superomedially-based in seven, laterally-based in eight, and superolaterally based in one. The tissue defects were the result of the excision of tumours in 18 patients, and of injury in three. The defects were located in the upper eyelid in two, the lower eyelid in five, the medial canthus in eight, the lateral canthus in four, and in the neighbouring orbitonasal zones in two. Different sized flaps were used according to the dimensions of the defects, which ranged from about 1 to 8 cm2. The upper eyelid flap is versatile, and provides tissue of ideal colour and texture. The procedure can be done quickly in one stage with minimal morbidity. PMID- 12625391 TI - Distortion and movement of the expander during skin expansion. AB - Distortion and movement of tissue expanders can cause expansion of the wrong area, such as the naevus or the scar that is to be resected. In 71 rectangular expanders, we examined the incidence of distortion (over 15 degrees) and movement (over 3 cm). We divided the expanders into three anatomical site groups: scalp, body, and extremities, and compared the complication rate between two study groups (distortion or movement, or not). In total, the incidence of distortion was 15/71 (21%) and that of movement 5/71 (7%). Distortion occurred mainly in the extremities (11/33,33%). The implanted expanders tended to move more often in the body part (3/15, 20%). In the extremities, the bigger the angle between the axis of the implanted expander and that of the extremity, the bigger the angle of distortion. Although the incidence of complications between the two groups was not significant, except for alteration in design of the flap, we recommend that these points should be considered when preoperative plans are being made for appropriate patients. PMID- 12625392 TI - Vacuum-assisted wound closure for cheaper and more comfortable healing of pressure sores: a prospective study. AB - Pressure sores are a common complication of patients with spinal injuries. The vacuum-assisted closure technique is widely used to induce and promote wound healing. We tested our clinical impression that pressure sores healed faster with vacuum-assisted closure, and compared it with the traditional wet-to-dry/wet-to wet technique with gauze soaked in Ringer's solution changed three times a day. Consecutive patients with pressure sores were entered into the study. Two randomised groups of 11 patients each with pressure sores of the pelvic region were included. We found no difference in time to reach 50% of the initial wound volume between the two methods. The vacuum-assisted group took a mean (SD) of 27 (10) days and the traditional group 28 (7) days. The two methods were equally effective in forming granulation tissue, so one can profit from the other advantages of the vacuum-assisted treatment (reduced costs and improved comfort) knowing that the effect on the formation of granulation tissue is as good as with the traditional treatment. PMID- 12625393 TI - Aesthetic considerations in the cervicofacial management of Madelung syndrome. AB - The most common concern in patients with Madelung syndrome who are seeking cosmetic improvement is the deposition of fat in the cervicofacial region. Surgical experience in a series of 15 patients was summarised to provide guidelines for a rational approach to a better aesthetic outcome. The records and clinical photographs of these patients were reviewed. Outcome was assessed in terms of adequacy of debulking, final overall symmetry, smoothness of contour, prominence of scar, and necessity for multiple sessions. Open excision was the preferred primary treatment. A long mid-neck, transverse skin crease incision provided good exposure and was superior to multiple local incisions. Other surgical details worth noting include subplatysmal dissection, thick skin flaps, dissection around instead of into lipomas, appropriate removal of redundant skin, adequate reduction of tumour size instead of total excision, haemostasis, suction drainage, and layered skin closure. Liposuction was a useful adjunct for contouring the sides of the jaws in selected patients. PMID- 12625394 TI - Isolation and in vitro cultivation of human urothelial cells from bladder washings of adult patients and children. AB - To acquire urothelial cells for in vitro engineering of urothelium, biopsy specimens were taken from the urological tract. In clinical practice the number of cells harvested by biopsy are limited and the procedure requires general anaesthesia in children. The purpose of this study was to find out if bladder washings from adult patients as well as children contained enough proliferative and colony-forming uroepithelial cells to regenerate urethral mucosa in vitro, and if the cells could be stored by freezing. Bladder washings from nine children and eight adult patients were collected from patients who were having procedures that required an indwelling catheter. All cultures grew colonies of cells with a morphological appearance typical for epithelial cell growth. The cultures could be expanded to confluent, stratified sheets, and cells that stained for pancytokeratin, indicating an epithelial origin. Cells stored in -150 degrees C could be cultured and expanded in vitro. No differences were seen between cells from adults and children. Bladder washing is a non-invasive way to obtain many autologous urothelial cells. The method is reproducible and well tolerated by children. The possibility of culturing cells obtained in this way into stratified grafts provides a unique way of reconstructing the urogenital tract by "tissue engineering". PMID- 12625395 TI - Magnetic resonance assessment of the double-crush phenomenon in patients with carpal tunnel syndrome: a bilateral quantitative study. AB - Our main goal was to assess the coexistence of narrowed cervical foramens and cervical canal stenosis in patients with carpal tunnel syndrome (CTS). We took magnetic resonance (MR) images of 120 wrists and 480 foramens in 60 age and sex matched subjects (30 patients with CTS and 30 controls). All patients had nerve conduction velocity tests before MR examination. We measured the volume of the carpal canal in all participants bilaterally. We also quantified the cross sectional areas of the cervical foramens on both sides from C4 to T1 and measured the diameter of the cervical canal in all participants. We searched for the prevalence and location of cervical spondylosis and disc prolapse on the side of the symptomatic wrist(s). All patients had CTS-related neurophysiological findings. There was no correlation between the symptoms and the reduced carpal canal volume. The mean (SD) area of the foramens was 109.5 (12.8) mm2 in the patients compared with the controls 126.4 (28.7) mm2 (p = 0.007). Cervical spondylosis and disc prolapse were more common in the patients than the controls at the C5-C6 and C6-C7 levels, and their locations were on the same side as the symptoms in the wrist(s) in 50% of cases. There was no difference in the size of the cervical canal between the two groups. The higher incidence of narrowed cervical foramens in the patients and its concordance with affected nerve roots on the same side as the CTS symptoms support the hypothesis of a double-crush phenomenon. PMID- 12625396 TI - Early use of artificial sensibility to improve sensory recovery after repair of the median and ulnar nerve. AB - Artificial sensibility based on use of a "tactile glove" which substitutes for lack of sensory afferent inflow with acoustic feedback, was used early after repair of the median and ulnar nerves in a 21-year-old man. After six and 12 months the functional outcome exceeded what is expected in adults, and analysis with calculations for the minimal detectable change (MDC) in tactile gnosis showed a true change. This case highlights the timing of sensory re-education after nerve repair and also emphasises the importance of early restitution of afferent inflow from a denervated hand during rehabilitation. PMID- 12625397 TI - Bifid median nerve: anatomical variant at the carpal tunnel. AB - We describe a rare case of a bifid median nerve in the wrist of a 4-year-old girl who presented with a laceration of her right hand. In addition to the nerve, the carpal tunnel was duplicated with a separate tunnel for each branch. PMID- 12625398 TI - The distally-based radial fasciosubcutaneous flap for soft tissue cover of the flexor aspect of the wrist. AB - The distally-based radial forearm fasciosubcutaneous flap is based on the distal perforators of the radial artery. We used it in a particularly difficult case involving loss of soft tissue at the wrist with exposure of tendons and nerves after an operation to section the transverse carpal ligament for carpal tunnel syndrome complicated by a chronic fistula. PMID- 12625399 TI - Diversity of Mannheimia haemolytica and pasteurella trehalosi serotypes from apparently healthy sheep and abattoir specimens in the highlands of Wollo, North East Ethiopia. AB - The prevalence and serotypic diversity of Mannheimia [Pasteurella] haemolytica and Pasteurella trehalosi from nasal swabs, sera and abattoir specimens from sheep in the highlands of Wollo, North East Ethiopia was investigated. Prevalence rates of 83% and 75% of these microorganisms were found in the serum samples and nasal swabs, respectively, from apparently healthy sheep. In a local abattoir, 205 lungs were investigated, 34% of which showed pneumonia, from which samples were collected from 51 lungs and the same number of corresponding tonsils. Mannheimia and Pasteurella species were isolated from 59% of these pneumonic lungs and 69% of the respective tonsils. M. haemolytica serotypes accounted for 41 (59%) and P. trehalosi for 11 (32%) of the isolates from the abattoir specimens. The majority (67%) of isolates from nasal swabs were P. trehalosi, M. haemolytica being isolated f rom 4 (13%) of the swabs. M. glucosida was isolated only from the tonsils. The predominant serotypes of the isolates from both the nasal swabs and the abattoir specimens were M. haemolytica A1 (17%) and P. trehalosi T4 (16%) and T3 (13%). P. trehalosi T15 was less commonly encountered, while M. haemolytica A9 and A13 were not isolated. Studies on sera from 100 sheep indicated that antibodies against M. haemolytica serotype A1 (14%) were most common, followed by A5 and A8 (each 10%) and A9 and P. trehalosi T3 (each 9%) and T4 (8%). Antibodies against M. glucosida or serotype All occurred in 2% of the sera. Multiple serotypes were common in all types of samples. The importance of including in vaccines the most prevalent serotypes involved in the pneumonia of sheep in the area is discussed. PMID- 12625400 TI - Biochemical profiles in Friesian cattle naturally infected with Theileria annulata in Saudi Arabia. AB - The biochemical parameters were studied in adult and young Friesian cattle naturally infected with Theileria annulatta in the Qassim Region, Saudi Arabia. Forty-three clinical cases of tropical theileriosis were studied, together with 40 clinically healthy Friesian cattle. Cattle clinically infected with T. annulata had significantly lower serum total protein, albumin, globulin, creatinine, calcium, phosphorus, magnesium, potassium, iron and copper concentrations and significantly higher AST activity and bilirubin concentration than the healthy cattle. PMID- 12625401 TI - Alternatives to blood as a source of DNA for large-scale scanning studies of canine genome linkages. AB - Participation and compliance are critical to the success of any large-scale study of canine disease using DNA markers. Most canine genetic studies rely upon DNA extracted from peripheral blood samples. We assessed the utility of buccal swab epithelial cells and toe nails as a source of DNA for use in genomic screening studies. Using eight multiplexed canine microsatellite markers, amplified DNA obtained from peripheral blood, and from freshly extracted buccal epithelial cells, and buccal swab DNA extracted and stored at 20 degrees C for 27 months or extracted from toe nails were compared for three dogs. The accuracy of the genotyping at each locus was identical for each preparation. Buccal swab DNA samples were readily and uniformly amplified and could be stored for years without loss of integrity. Each buccal swab provided sufficient DNA for more than 200 individual PCR reactions. Toe nails provided ample DNA for thousands of PCR reactions and had the added advantage of ease of storage of the original tissues. These studies demonstrate the potential utility of DNA derived from buccal swabs or nails in large-scale genomic scanning and marker linkage studies. PMID- 12625402 TI - Immunosuppression in broiler chicks fed aflatoxin and inoculated with fowl adenovirus serotype-4 (FAV-4) associated with hydropericardium syndrome. AB - A total of 240 unvaccinated day-old broiler chicks, which had been found to be negative for antibodies against FAV-4, were divided into four groups of 60 chicks each. Group A was fed aflatoxin at 1 ppm from 7 days to 7 weeks of age. Group V was infected intra-abdominally at 14 days of age with 0.2 ml of FAV-4, having a titre of 10(5.5) TCID50 per 0.2 ml. The combined group AV was given the aflatoxin and infected with FAV-4. The fourth group C served as the control. More pronounced clinical signs, a higher mortality rate (56.7%), and reductions in body weight gain and in the organ to body weight ratios of the bursa and spleen were recorded in group AV. A significant (p < 0.01) reduction in the HI antibody titre following vaccination against Newcastle disease, and of skin thickness in the delayed hypersensitivity test following sensitization with DNCB, indicated an additive immunosuppressive effect from aflatoxin and FAV-4 on the humoral and cell-mediated immune responses in group AV compared to groups A and V. Microscopically, marked depletion and degeneration of lymphocytes in the thymus, bursa, spleen and caecal tonsils were observed in group AV up to 5 weeks PI. PMID- 12625403 TI - Experimental Tribulus terrestris poisoning in sheep: clinical, laboratory and pathological findings. AB - Eleven native sheep, 1-2 years old, of both sexes were randomly divided into two groups, 6 sheep being allocated to the experimental group and 5 serving as controls. The sheep in the experimental group were fed 80% Tribulus terrestris and 20% alfalfa hay and wheat straw, while the control sheep were given a mixture of 40% alfalfa hay and 60% wheat straw. Clinical signs of hepatogenous photosensitivity were observed from day 11, including reddening and crust formation on the muzzle, nose, ears and eyelids, depression, weight loss, icterus, conjunctivitis, and yellow discoloration of the urine. Laboratory findings on weekly samples indicated significant differences (p < 0.05) in white blood cell count, total plasma protein and fibrinogen, total and direct bilirubin, blood urea nitrogen and creatinine concentrations, and aspartate aminotransferase and alkaline phosphatase activities. There were no significant differences in the packed cell volume, in the neutrophil, lymphocyte or eosinophil counts, or in the serum calcium, phosphorus, potassium, sodium or chloride concentrations. At necropsy of the experimental animals, there were various degrees of generalized icterus and the livers were swollen and discolored by bile pigment. Histopathological examination revealed varying amounts of crystalloid material in the bile ducts and renal tubules, hepatocellular degeneration, biliary fibrosis and proliferation, renal tubular necrosis and focal necrosis of cardiac muscle. PMID- 12625404 TI - Use of the reverse transcription polymerase chain reaction (RT-PCR) for the rapid diagnosis of foot and mouth disease in South America. AB - Foot and mouth disease (FMD) is a limiting factor for the economic progress of the animal industry in South America. The presence of the disease results in the imposition of national and international sanitary barriers to animals and animal products, and, most especially, a reduction in the availability of protein from animal origin and in income. Rapid and accurate identification of infected animals, those with either clinical or subclinical disease as well as with persistent infection, is essential for maintaining an efficient eradication programme. The polymerase chain reaction was used to rapidly identify infected animals. With a primer set that corresponds to a conserved region of the 3D sequence of the viral genome, it was possible to amplify, regardless of the serotype, 116 strains of FMD virus, of which 109 were strains collected from outbreaks of FMD throughout South America from 1945 to the most recent outbreaks in 2000/2001. The PCR technique should be of considerable value in facilitating the diagnosis of FMD in South America. where laboratory resources are limited and a rapid response is needed, particularly in areas where national programmes for controlling or eradicating the disease are being implemented. PMID- 12625405 TI - Avian reovirus induces an inhibitory effect on lymphoproliferation in chickens. AB - The cellular immune responses of chickens inoculated with the vaccine strain S 1133 and/or a field isolate VA-1 of avian reovirus (ARV) were studied. Both strains of virus caused inhibition of the phytohaemagglutinin (PHA)-induced lymphoproliferative response of peripheral blood mononuclear cells (PBMC) and splenic mononuclear cells (SMC) during the initial stage from day 4 up to day 10 post-inoculation (PI), with a later return to the normal value. The inhibition in the PHA-induced lymphoproliferation of SMC could be partially overcome by depletion of adherent cells. The supernatant of the PHA-stimulated SMC culture was also checked in vitro for the presence of suppressive factor(s) produced in response to ARV infection. The culture supernatant from chickens at day 5 PI caused significant inhibition of the PHA-induced lymphoproliferation of control birds, suggesting the presence of suppressive factor(s). ARV infection also significantly inhibited IL-2 production on day 5. There was a significant increase in nitric oxide production by the splenic mononuclear cells of chickens inoculated with either strain of ARV. PMID- 12625406 TI - A case of autoimmune hypophysitis associated with Graves' disease. AB - In a 76-year-old woman with hyperthyroidism, hyperprolactinemia and thickening of the pituitary stalk on magnetic resonance imaging (MRI) was presented. Thyroid stimulating antibody (TSAb) was positive and anti-pituitary antibodies against 49 and 68 kD human anterior pituitary membrane antigens were detected immunologically. Secretion of pituitary hormones was almost normal except for suppressed TSH and hyperprolactinemia. As autoimmune etiologies were likely involved in the disorders, autoimmune hypophysitis associated with Graves' disease was arrived at as the plausible diagnosis. PMID- 12625407 TI - Changes in telomerase activity in experimentally induced atretic follicles of immature rats. AB - Follicular atresia is characterized by apoptosis of granulosa cells, and telomerase plays an important role in the apoptotic process. To study the relationship between follicular atresia and telomerase activity, we investigated changes in telomerase activity and localization in experimentally induced atretic follicles of immature rats. Immature female Sprague-Dawley rats received 15 IU equine CG (eCG) by subcutaneous injection. Rats were killed under ether anesthesia at 2, 3, 4, or 5 days after eCG injection. Telomerase activity in granulosa cells was measured by telomeric repeat amplification protocol (TRAP) assay, and telomerase localization in the ovary was examined by in situ TRAP assay. Telomerase activity was detected at high levels in granulosa cells on day 2 after eCG injection regardless of follicle size, and levels were significantly decreased in large follicles, with atretic changes, on days 4 and 5. No such decrease was observed in granulosa cells of small follicles. In the next experiment, rats received subcutaneous injections of estradiol (1, 10, or 50 microg/rat) to prevent follicular atresia or sesame oil as a control from day 2 to day 4 after eCG injection and were killed under ether anesthesia on day 5 after eCG injection. The changes observed in the large follicles on days 4 and 5 in oil treated rats were not observed with estradiol treatment. These findings suggest that the telomerase in granulosa cells is likely to play an important role for healthy follicle life and that loss of its activity may be associated with follicular atresia. PMID- 12625408 TI - Adult growth hormone deficiency in Japan: results of investigation by questionnaire. AB - Adult GH deficiency (AGHD) has been established as a syndrome associated with various metabolic disturbances such as hyperlipidemia, impaired glucose tolerance and protein catabolism, in addition to changes in body composition such as increased visceral fat, decreased muscle mass and bone density. We investigated the clinical findings, complications and prognosis of AGHD in Japan. The questionnaire was sent to various expert facilities of endocrinology and metabolism to gather cross-sectional information as well as longitudinal follow up data on adult patients with hypopituitarism. We received answers on 422 subjects, of which number the GH stimulation test was performed in only 63% of them. An age- and sex-matched group of 259 adults with hypopituitarism (125 male and 134 female subjects) was finally selected for this investigation. Of them 185 subjects (81 male and 104 females) were diagnosed as AGHD with plasma peak GH levels less than 3 ng/ml after GH stimulation test. Male adult patients with GHD had significantly lower ratio of smoking and drinking in their life style compared with those without GHD. Male adult patients with GHD revealed significantly higher BMI on physical examination, and significantly higher plasma ALT, AST, total cholesterol, and LDL cholesterol in blood chemistry compared with those without GHD (P < 0.05). Though patients with ischemic heart disease were more frequent in female patients than male patients, the rate of frequency was not different between female adult patients with and without GHD. Clinical characteristics found in especially male adult patients with GHD in Japan were consistent with findings reported so far in foreign countries. However, consequent complications such as atherosclerosis seemed less severe than expected. Moreover, GH stimulation test for the diagnosis of AGHD as well as clinical test to perform when AGHD was suspected is still less frequently carried out. Therefore, the clinical outcome of AGHD in our country requires further investigation. PMID- 12625409 TI - A case of lymphocytic infundibuloneurohypophysitis associated with systemic lupus erythematosus. AB - A 27-year-old man was admitted to our hospital with facial erythema and general malaise. He had previously suffered from orbital myositis, central diabetes insipidus (DI), peripheral neuritis, and hypogonadotropic hypogonadism. Physical and immunological examinations revealed that he was suffering from systemic lupus erythematosus (SLE). Magnetic resonance imaging of the hypothalamic-pituitary region demonstrated a significant enlargement of the pituitary stalk and posterior pituitary. Endocrinological examinations showed that he had not only DI and hypogonadotropic hypogonadism but also hypoadrenalism and hypothyroidism, which were ascribed to the pituitary stalk lesion. Lymphocytic infundibuloneurohypophysitis associated with SLE was diagnosed. Administration of 30 mg/day of prednisolone for one month resulted in a marked reduction of the pituitary stalk thickening and posterior pituitary. It is recommended that a pharmacological dose of glucocorticoid be used in the treatment of lymphocytic hypophysitis patients who show significant thickening of the pituitary stalk and/or a large pituitary mass. PMID- 12625410 TI - Effects of growth hormone on testicular dysfunction induced by cyclophosphamide (CP) in GH-deficient rats. AB - Growth hormone (GH) is known to accelerate spermatogenesis and maintain gonadal function. In this study, we evaluated the effect of GH on recovery from testicular damage induced by cyclophosphamide (CP). Eleven- to fourteen-week-old GH-deficient Lewis rats (dw/dw) were divided into 4 groups (n = 10 each), with one group serving as controls. In the CP group, CP was intravenously administered in daily doses of 50 mg/kg for 2 days, followed by daily doses of 10 mg/kg for the next 3 days. In the GH group, rat GH was subcutaneously administered at a daily dose of 0.3 mg/kg until the rats were sacrificed. In the CP/GH group, GH and CP administration were started simultaneously. In the CP/preGH group, GH administration was started 14 days before CP administration. Five rats from each group were sacrificed at days 14 and 28 after administration of CP. Spermatogenesis was then evaluated morphometrically by counting numbers of cells at several stages of the spermatogenic cycle. On day 14, there were no significant differences in the numbers of the spermatocytes between CP and CP/GH group. On day 28, the numbers of spermatocytes and motility of spermatozoa in CP/GH group were greater than those of CP group were. In the CP/preGH group, these effects of GH administration were not observed. These results suggested that administration of GH improved testicular function damaged by CP under GH deficient condition, when GH and CP administration are started simultaneously. PMID- 12625411 TI - Hypothyroidism as a cause of rhabdomyolysis. AB - We describe a patient presenting with muscular symptoms and rhabdomyolysis without any other precipitating factor, except primary hypothyroidism. After thyroxine replacement, musculoskeletal symptoms disappeared and creatine kinase concentrations decreased. Hypothyroidism is a rare cause of rhabdomyolysis, but should always be considered in a patient with an unexplained increase in creatine kinase concentrations. PMID- 12625412 TI - A novel missense mutation of AIRE gene in a patient with autoimmune polyendocrinopathy, candidiasis and ectodermal dystrophy (APECED), accompanied with progressive muscular atrophy: case report and review of the literature in Japan. AB - Autoimmune polyendocrinopathy, candidiasis, and ectodermal dystrophy (APECED) also known as autoimmune polyglandular syndrome type I, is a rare autosomal recessive disorder that results in several autoimmune diseases due to mutations in the AIRE (autoimmune regulator) gene. A 39-year-old female patient developed chronic mucocutaneous candidiasis at 3 yrs, idiopathic hypoparathyroidism at 11 yrs, chronic hepatitis at 23 yrs, Addison's disease and diabetes mellitus type I at 27 yrs. In addition, the patient developed progressive muscular atrophy of unknown etiology at the beginning of the third decade, and is bedridden at the present time. Her grandparents, parents, brother and daughter did not develop any features of APECED, but her father died of hepatoma. Direct sequencing of the AIRE gene revealed a novel missense mutation at exon 1 (R15C), which was identified to be of maternal origin. The other mutation was not found despite repeated sequencing of the whole coding regions. The R15C mutation was not detected in patients with idiopathic hypoparathyroidism (N= 10), idiopathic Addison's disease (N = 3), and normal subjects (N = 55). Although we could not analyze the father's gene, these results suggest that the patient is probably a compound heterozygote of the AIRE gene, in which the other abnormal allele could not be identified by the present analytical method. These data are compatible with the recent review that only one defective allele was detectable in some patients with clinically evident APECED. We found only six Japanese patients compatible with diagnosis of APECED, indicating that this autoimmune disease is extremely rare in our country. PMID- 12625413 TI - Pituitary adenomas in adolescent patients with multiple endocrine neoplasia type 1. AB - Two juvenile patients with multiple endocrine neoplasia type 1 (MEN1) who developed pituitary adenomas are reported. The first case, a 14-year-old girl, developed prolactinoma and manifested delayed puberty and growth arrest. The second case, a 16-year-old boy, was asymptomatic and a pituitary adenoma accompanied by mild elevation of PRL and GH was identified through family screening. His growth and pubertal development was not impaired. Medication with bromocriptine was started for both cases with good therapeutic responses. These cases emphasize relevance of early screening of endocrine disorders for members of families with MEN1. PMID- 12625414 TI - Prospective examination of adrenocortical function in advanced AIDS patients. AB - In human immunodeficiency virus infected individuals, human cytomegalovirus (CMV) remains a significant pathogen. The adrenal gland is a preferential site of CMV disease in acquired immunodeficiency syndrome (AIDS) patients. However, glucocorticoid replacement is often not selected because of the risk of exacerbating underlying infection. To evaluate the need for glucocorticoid replacement in these patients, we performed a prospective study to investigate the adrenal function in 60 advanced AIDS patients. Their adrenal function including rapid ACTH test (RAT), basal plasma ACTH level, and daily urinary free cortisol level was evaluated. Approximately 25% of the patients turned out to be abnormal in this evaluation. Almost 60% of the patients could be followed up for one year or until their death. Using the follow-up data, we calculated sensitivity, specificity, positive predictive value and negative predictive value. Excretion of urinary free cortisol with normal RAT was comparable to normal controls, whereas patients with abnormal RAT excreted significantly lower urinary free cortisol. During hospitalization, 14 patients with normal RAT had febrile episode. During the febrile period the concentration of the urinary free cortisol level increased by 2.2 times. This study suggests that glucocorticoid replacement is necessary for AIDS patients suspected as clinically adrenal insufficiency, and that the dose of glucocorticoid replacement might be increased during sick days in AIDS patients with abnormal adrenal function. PMID- 12625415 TI - The effects of nateglinide following oral glucose load in impaired glucose tolerance subjects: rapid insulin stimulation by nateglinide in IGT subjects. AB - This study was designed to determine the effect of a novel insulin secretagogue, nateglinide, on the glycemic response curve and early insulin secretion following oral glucose load in impaired glucose tolerance (IGT) subjects. Thirteen subjects were given a 75 g oral glucose tolerance test (75 g OGTT), the findings of which resulted in the diagnosis of IGT. The subjects returned to our hospital immediately. Eight subjects, in whom neither body weight nor life style (daily diet and exercise) was significantly altered during this period, were given 90 mg of nateglinide 5 min before a second oral glucose load in order to examine restoration of impaired early insulin secretion. Nateglinide administration resulted in the almost normalization of the glycemic response curve with restoration of impairment in early insulin response at 30 and 60 min after an oral glucose load. The area under the secreted insulin-time curve was not changed significantly by nateglinide administration. A single dose of nateglinide was shown to almost normalize the glycemic response curve after a 75 g OGTT and to restore impairment in early insulin response in IGT subjects. PMID- 12625416 TI - Association of HLA-CW3 with type 1 diabetes mellitus and idiopathic hypoparathyroidism in the Japanese. PMID- 12625417 TI - Conjugated linoleic acids: all the same or to everyone its own function? AB - Conjugated linoleic acid (CLA) is a generic term referring to a mixture of geometrical and positional isomers of linoleic acid in which up to 16 members have been identified. Many potentially beneficial health effects have been ascribed to these fatty acids when consumed as a mixture, and where generally 2 isomers dominate, e.g. the 9c, 11t-isomer, the so-called rumenic acid, and the 10t, 12c-isomer: anti-carcinogenic, immune modulator, anti-atherosclerotic, and anti-obesity among the most spectacular. The question arises as to whether the pleiotropic biological activity is supported by one or several of the isomers. Recent studies using pure individual isomers have started to elucidate this issue, but many others are required to ascribe a respective role to each CLA isomer (the main ones as well as the minor ones), such as those occurring in some complex mixtures already commercially available, or even in foodstuff. The aim of the present study was to focus on the CLA-isomer specific effects depicted in the literature up to now. PMID- 12625418 TI - Rumen escape of methionine and lysine administered intraruminally to growing double-muscled Belgian Blue bulls. AB - In many dietary conditions, methionine (Met) and lysine (Lys) are the most limiting amino acids (AA) for ruminants. The AA protected from ruminal fermentation are not commercially available, with the exception of Met which is not always economical, especially for meat production. This study measured ruminal escape of free Met and Lys supplemented intraruminally to fast growing bulls. Six double-muscled Belgian Blue bulls, fed a high concentrate diet and fitted with a rumen cannula, received free Met (40 g x d(-1)) and free Lys (60 g x d(-1)), individually or simultaneously, in a duplicated Latin square design. The mean ruminal escape of Met and Lys reached 37 and 45% respectively, and did not differ if administered separately or together. Plasma Lys and Met concentrations were increased by 504 and 126%, respectively. Substantial proportions of free AA escaped ruminal fermentation and were available for absorption from the small intestine when they were administered at physiologically high levels. PMID- 12625419 TI - Nitrogen utilisation by dairy cows fed diets differing in crude protein level with a deficit in ruminal fermentable nitrogen. AB - We studied the efficiency of nitrogen utilisation by dairy cows, using three diets differing in the crude protein (CP) level but with similar deficits (10 g x kg(-1) dry matter, DM) in ruminal fermentable nitrogen. There was no difference in milk yield from the cows offered the three diets (130, 145 and 160 g CP x kg( 1) DM). The milk protein content differed between the two most extreme diets (28.9 vs. 29.9 g x kg(-1), P < 0.05), resulting in higher protein yields for the highest CP treatment (P < 0.01). The efficiency of nitrogen utilisation, calculated as the proportion of ingested nitrogen recovered in the milk, was significantly higher for the 130 g CP x kg(-1) DM diet than for the other two diets (0.37 vs. 0.33 and 0.32 respectively. P < 0.01). The different diets also resulted in different levels of nitrogen excretion into the environment (237, 270 and 330 g N x d(-1), P < 0.01). Hepatic deamination of the amino acids may have generated additional energy to enable the animal to make use of the additional nitrogen in the diet, resulting in an increase in plasma urea concentration. PMID- 12625420 TI - Protein degradation in the rumen of red clover forage at various stages of growth and conserved as silage or wrapped big bales. AB - In order to study the extent to which rumen soluble nitrogen can contribute to the intestinal flow, a study was carried out to simultaneously assess the dynamics of protein disappearance from dacron bags placed in the rumen and the amount of various N products in the rumen fluid (total nitrogen (tN), ammonia nitrogen (NH3-N), non-ammonia nitrogen (NAN)). The measurements were carried out on 4 sheep fed successively various red clover forages. These forages included the initial growth of fresh red clover (50% bud, first flower, and full flower). In addition, one silage and one wrapped big bale at the first flower stage and two wrapped big bales (harvested at 51% and 71% dry matter) at the full flower stage were given. The effective degradability of nitrogen (DegN) for a fresh forage estimated from the nylon bag procedure did not vary (p > 0.05) with the vegetation stage (0.727 for the bud stage, 0.694 at the first flower, 0.706 at the full flower). The DegN of the silage was higher (p < 0.05; 0.735) and the DegN of the wrapped big bale was markedly lower (p < 0.05; 0.660), than the original fresh forage at the first flower. The DegN of the wrapped big bales made at 51 and 71% DM, respectively, were 0.625 and 0.604 against 0.706 for fresh forage at the full flower stage. The concentrations of tN and NAN in the rumen fluid were low, highest 1 h to 2 h after feeding, and then decreasing up to 7 h after feeding whatever the growth stage and conservation mode. A part of the solubilised nitrogen remained as protein 1 h after feeding for fresh red clover harvested at various growth stages, while minimal protein could be seen in the rumen fluid after the sheep were fed silage or wrapped big bales. The part of NAN escaping rumen degradation and transiting with the rumen fluid was between 7 and 13% of the nitrogen disappearing from the nylon bags (NAN/CP x DegN) placed in the rumen. There was only a small difference for forages at different stages of growth, or modes of conservation. This fraction was higher for wrapped big bales and particularly for the late stage forage (wrapped big bale, 71% DM, harvested at the full flower stage). PMID- 12625421 TI - The pesticide methoxychlor given orally during the perinatal/juvenile period, reduced the spermatogenic potential of males as adults by reducing their Sertoli cell number. AB - Perinatal and juvenile oral treatment of rats with the insecticide, methoxychlor (MXC), reduced testicular size and other reproductive indices including the number of epididymal spermatozoa in those animals as adults 161. The objective was to determine if these males exposed during development had fewer Sertoli cells which might explain these testicular effects. Rat dams were gavaged with MXC at 0, 5, 50, or 150 mg x kg(-1) x day(-1) for the week before and after they gave birth. Resulting male pups (15/group) then were dosed directly from postnatal day 7 to 42. Testes were fixed in Bouin's and in OsO4, embedded in Epon and sectioned at 0.5 microm, stained with toluidine blue, and evaluated stereologically or cut at 20 microm to measure Sertoli cell nuclei with Nomarski optics. Sertoli cell number was calculated as the volume density of the nucleus times the parenchymal weight (90% of testicular weight) divided by the volume of a single Sertoli cell nucleus. Across dose groups, there were no changes in the nuclear volume density, the volume of a single nucleus, or the number of Sertoli cells per g parenchyma. There were highly significant dose-related changes in the volume of Sertoli cell nuclei per testis and the number of Sertoli cells per testis. Reduced testicular weight (r = 0.94) and reduced numbers of epididymal spermatozoa (r = 0.43) were significantly (p < 0.01) correlated to reduced number of Sertoli cells per testis. Hence, perinatal and juvenile oral exposure to MXC can reduce spermatogenic potential of males as adults by reducing their number of Sertoli cells. PMID- 12625422 TI - Successful pregnancies in cows following double freezing of a large volume of semen. AB - The objective of the following paper is to describe a new technology for large volume and double freezing of semen in 12 mL test tubes. Semen from two different bulls was frozen with a new technique using 12 mL test tubes and was refrozen after thawing in mini straws. All freezing was done in a "Multi thermal gradient" (MTG) freezing apparatus, which moves the container at a constant velocity (V) through a thermal gradient (G) producing a controlled cooling rate B = (G) x (V). Each of the two bulls ejaculated were evaluated for post thaw motility in the lab and then in a field trial which was carried out in a split sample mode. We inseminated 105 cows after a double freezing/thawing cycle, and another 123 cows were inseminated with semen frozen in mini-straws and a conventional method. The results showed a 75 +/- 5% post thaw motility after freezing a 12 mL test tube and 50 +/- 5% after a second freezing/thawing in mini-straws, respectively. Controlled vapour freezing showed a 60 +/- 10% post thaw motility. The results of the field trial showed a pregnancy rate of 44% (47/105) for the double freezing group in comparison to 45.5% (56/123) for the controlled group. These results can be beneficial for large volume freezing, and therefore for bull semen cryobanking in a large volume which will be followed by second freezing in a regular insemination volume. PMID- 12625423 TI - Superovulation in cattle: practical aspects of gonadotropin treatment and insemination. AB - Embryo transfer (ET) in cattle has been used for the realisation of breeding programmes world-wide for more than 20 years. The efficiency of breeding technology, i.e. the breeding progress and costs, depends to a large extent on the results of superovulatory treatment and artificial insemination (A.I.). The results of this step are characterised by a high degree of variation. Numerous attempts have been undertaken to explain the reason(s) for this. Numerous attempts have also been made to clarify the importance of different factors affecting the results. Undoubtedly, the applied hormones and the scheme of insemination itself are main factors, which influence the number and the portion of transferable embryos. Therefore this paper is focused on the following aspects of superovulatory treatment with FSH: dose-response relations, bioactivity of the glycoprotein, FSH/LH ratio, ovulation time and time-oriented insemination, frequency of gonadotropin administration and follicular population at the time of gonadotropin application. PMID- 12625424 TI - Recent advances in the superovulation in cattle. AB - The variability in the superovulatory response continues to be one of the most frustrating problems with embryo transfer in cattle. The removal of LH from pituitary extracts has tended to reduce the variability in response, and several studies involving the use of the purified porcine pituitary extract. Folltropin-V are reviewed. The major source of variability in the superovulatory response in cattle is the status of ovarian follicles at the time of initiation of gonadotrophin treatments. Data support the benefits of initiating gonadotrophin treatments at the time of emergence of a follicular wave. Incorporation of techniques designed to control follicular wave dynamics, such as follicular ablation, or treatment with estradiol/progesterone, have reduced the variability caused by treating cows at different stages of follicular development, and at the same time improved response by taking advantage of endogenous recruitment and selection mechanisms. New protocols offer the convenience of being able to initiate gonadotrophin treatments quickly and at a self-appointed time, without the necessity of estrus detection and without sacrificing response. Methods can be used for repeated superstimulation of donor animals at 25 to 30 day intervals, without regard to estrus detection or stage of the estrous cycle, and without compromising embryo production. PMID- 12625425 TI - The monitoring of bovine pregnancies derived from transfer of in vitro produced embryos. AB - Both an increased rate of embryonic, foetal and perinatal losses, and the occurrence of deviations in foetal and placental development are associated with bovine pregnancies obtained from in vitro produced embryos. This thus requires for a more accurate and frequent monitoring of foetal and maternal functions during pregnancies. Such approaches will enable to establish the period during which these losses and deviations in development occur and to plan possible clinical interventions. This paper reviews some recent data on return rates, late embryonic and foetal losses in recipients after the transfer of either MOET, IVF or nuclear transfer embryos. Special attention is paid to the diagnostic value of measurements of pregnancy specific/associated proteins and progesterone in maternal plasma. Possibilities to measure foetal body sizes, size of placentomes and foetal heart rate by means of transrectal or transabdominal ultrasonography are illustrated with data from the literature and with recent results from our own large field study with MOET, IVP-co-culture and IVP-SOF embryos. PMID- 12625426 TI - Rethinking confrontation in alcohol and drug treatment: consideration of the clinical context. AB - The role of confrontation in the treatment of substance misuse problems is widely debated in the field. Historically, confrontation of denial has been important in treatment, particularly in residential programs. However, confrontation has also been criticized as counterproductive. Miller et al. colleagues have found that confrontation from treatment professionals is correlated with poor outcome. This paper reviews the literature supporting and criticizing confrontation. It is suggested that confrontation is not a monolithic concept. Rather, it is contingent upon clinical contextual factors that current research has not addressed, such as the treatment setting, treatment philosophy, client characteristics, the client's view about their relationship with confronter, the extent to which the client views the confrontation as legitimate, the client's perception of the confronter's motivation, and the level of emotional intensity involved in the confrontive exchange. Confrontation from multiple sources requires more focus, including confrontation from treatment peers, family members, the workplace, the legal system, social welfare workers, and medical professionals. PMID- 12625427 TI - Normalization of marijuana use: its effects on adolescents' intentions to use marijuana. AB - The comparative salience of attitudes, normative influences, and perceptions of the ease or difficulty of engaging in marijuana use was examined in 91 adolescents from three subsamples: those living in Nimbin, Australia, where marijuana use appears to have become normalized, a control group, and a group of homeless adolescents, surveyed in 1997. There were no differences between two of the groups in relation to their attitudes towards marijuana, and no differences in perceived access to the drug. For those who intended using marijuana, personal beliefs were significantly more salient than beliefs about what important others thought they should do. PMID- 12625428 TI - Social problems, psychological well-being, and childhood parenting experiences in a community sample of heroin addicts in central London. AB - Well-validated instruments were used to compare the social adjustment, psychological well-being, and childhood parenting experiences of 105 heroin dependent people and a matched sample of 103 unemployed people recruited from April to November 2001 via newspaper advertisements and word of mouth in central London. The groups were not homogenous. Both groups had more social and psychological problems than population norms. The unemployed group reported significantly more social and interpersonal problems than the heroin users although they had significantly fewer psychological problems. Childhood parenting experiences were comparable between both groups suggesting this is not a specific cause of heroin dependence. PMID- 12625429 TI - Psychostimulant dependence in a community sample. AB - OBJECTIVE: To examine the prevalence of psychostimulant dependence and the characteristics associated with nonmedical users' development of dependence. METHODS: The study sample was drawn from the 1995 to 1998 National Household Surveys on Drug Abuse. Statistical analysis was conducted on a total of 1047 individuals aged 12 or older who reported nonmedical use of stimulants in the past year. Multiple multinomial logistic regression identified factors related to stimulant dependence and dependence problems. RESULTS: Among all past year stimulant users, 19% met criteria for stimulant dependence in the past year, and an additional 16% reported having one to two dependence problems. Adjusting for demographics and drug use characteristics, female stimulant users were an estimated 2.6 times more likely than male users to develop dependence. Not only did the Western region of the United States have more recent stimulant users than other regions, its users also were more likely to meet criteria for dependence or experience dependence problems. Stimulant users who had increased odds of progressing into dependence were characterized by an early onset of stimulant use, coexisting multiple illicit drug use, and an onset of daily cigarette smoking between the ages of 13 and 17 years. CONCLUSIONS: Gender differences in initial stimulant use and progression to dependence require further investigation, including contextual, cultural, or perceptual factors related specifically to the choice of drugs by females. PMID- 12625430 TI - Childhood sexual abuse, PTSD, and the functional roles of alcohol use among women drinkers. AB - The present study investigated whether a history of childhood sexual abuse (CSA) or current post traumatic stress disorder (PTSD) was associated with alcohol related expectancies and recalled effects of drinking reported by women in substance user treatment. The results indicate that CSA status was not associated with the alcohol-related expectancies or effects of drinking reported by women in substance user treatment. However, those currently manifesting PTSD were more likely than their peers without PTSD to report greater alcohol-related expectancies of tension reduction and having experienced greater positive enhancement from drinking. Theoretical and clinical implications of these findings are considered. PMID- 12625431 TI - The drug-user husband and his wife: attachment styles, family cohesion, and adaptability. AB - This study which assesses the association between the attachment styles of drug user husbands (n = 56) and their wives (n = 56) and their perceptions of family dynamics was conducted in 1998. The population study included heroin (52.9%) and multidrug detoxified outpatients. All subjects completed the Adult Attachment Style Scale and the FACES III. Results indicated that the perceptions of family adaptability and cohesion among the drug-user husbands and their wives did not differ from the Israeli norm. Most of the drug users (60.7%) were characterized by the avoidant attachment style, followed by the secure style (26.8%), and the anxious/ambivalent style (12.5%). Half the wives (53.6%) were characterized by the secure style, followed by the avoidant style (42.9%) and the anxious/ambivalent style (3.6%). A secure style in husband and wife was associated with higher levels of family cohesion and adaptability, and the anxious/ambivalent style with a lower perception of family cohesion and adaptability. These findings have important implications for rehabilitation prospects and for planning intervention programs. PMID- 12625432 TI - Construct validity for alcohol dependence as indicated by the SUDDS-IV. AB - OBJECTIVE: This study considers the diagnostic construct validity of the DSM-IV (Diagnostic and Statistical Manual-lV) for "alcohol dependence". Previous reports have indicated that "dependence" constitutes a more distinct and pronounced syndrome than "alcohol abuse". METHOD: Data were collected in 2000-2001 on 1340 male and female inmates evaluated for "substance use disorders" using the SUDDS IV, a detailed structured diagnostic interview, to collect data on all of the DSM IV diagnostic criteria for "abuse" and "dependence". RESULTS: Dependent individuals tended to produce distinct and extensive symptom profiles that distinguished them from individuals without a diagnosis or those meeting abuse criteria. CONCLUSIONS: Alcohol dependence as defined by the DSM-IV appears to be quite distinct from abuse and can be identified unequivocally for the majority of dependent cases. PMID- 12625433 TI - School-based early intervention and later child maltreatment in the Chicago Longitudinal Study. AB - Investigated were the effects of participation in the Title I Child-Parent Centers (CPC) on substantiated reports of child maltreatment for 1,408 children (93% of whom are African American) in the Chicago Longitudinal Study. The CPCs provide child education and family support services in high-poverty areas. After adjusting for preprogram maltreatment and background factors, 913 preschool participants had significantly lower rates of court petitions of maltreatment by age 17 than 495 children of the same age who participated in alternative kindergarten interventions (5.0% vs. 10.5%, a 52% reduction). Participation for 4 to 6 years was significantly associated with lower rates of maltreatment (3.6% vs. 6.9%, a 48% reduction). Findings based on child protective service records (as well as combined protective service and court records) were similar. Preschool length, family risk, and school poverty were associated with lower rates of maltreatment. Parental involvement in school and school mobility were significant mediators of intervention effects. PMID- 12625434 TI - Properties of school Chinese: implications for learning to read. AB - The properties of the 2,570 Chinese characters explicitly taught in Chinese elementary schools were systematically investigated, including types of characters, visual complexity, spatial structure, phonetic regularity and consistency, semantic transparency, independent and bound components, and phonetic and semantic families. Among the findings are that the visual complexity, phonetic regularity, and semantic transparency of the Chinese characters taught in elementary school increase from the early grades to the later grades: Characters introduced in the 1st or 2nd grade typically contain fewer strokes, but are less likely to be regular or transparent, than characters introduced in the 5th or 6th grade. The inverse relation holds when characters are stratified by frequency. Low-frequency characters tend to be visually complex, phonetically regular, and semantically transparent whereas high frequency characters tend to be the opposite. Combined with other findings, the analysis suggests that written Chinese has a logic that children can understand and use. PMID- 12625435 TI - Children's suggestibility in relation to their understanding about sources of knowledge. AB - In the experiments reported here, children chose either to maintain their initial belief about an object's identity or to accept the experimenter's contradicting suggestion. Both 3- to 4-year-olds and 4- to 5-year-olds were good at accepting the suggestion only when the experimenter was better informed than they were (implicit source monitoring). They were less accurate at recalling both their own and the experimenter's information access (explicit recall of experience), though they performed well above chance. Children were least accurate at reporting whether their final belief was based on what they were told or on what they experienced directly (explicit source monitoring). Contrasting results emerged when children decided between contradictory suggestions from two differentially informed adults: Three- to 4-year-olds were more accurate at reporting the knowledge source of the adult they believed than at deciding which suggestion was reliable. Decision making in this observation task may require reflective understanding akin to that required for explicit source judgments when the child participates in the task. PMID- 12625436 TI - Do beez buzz? Rule-based and frequency-based knowledge in learning to spell plural -s. AB - There has been much discussion about whether certain aspects of human learning depend on the abstraction of rules or on the acquisition of frequency-based knowledge. It has usually been agreed, however, that the spelling of morphological patterns in English (e.g., past tense -ed) and other languages is based on the acquisition of morphological rules, and that these rules take a long time to learn. The regular plural -s ending seems to be an exception: Even young children can spell this correctly, even when it is pronounced /z/ (as in bees). Reported here are 3 studies that show that 5- to 9-year-old children and adults do not usually base their spellings of plural real-word and pseudo-word endings on the morphological rule that all regular plurals are spelled with -s. Instead, participants appeared to use their knowledge of complex but untaught spelling patterns, which is based on the frequency with which certain letters co-occur in written English. PMID- 12625437 TI - The neuropsychology of Down syndrome: evidence for hippocampal dysfunction. AB - This study tested prefrontal and hippocampal functions in a sample of 28 school aged (M = 14.7 years, SD = 2.7) individuals with Down syndrome (DS) compared with 28 (M = 4.9 years, SD = .75) typically developing children individually matched on mental age (MA). Both neuropsychological domains were tested with multiple behavioral measures. Benchmark measures of verbal and spatial function demonstrated that this DS sample was similar to others in the literature. The main finding was a significant Group x Domain interaction effect indicating differential hippocampal dysfunction in the group with DS. However, there was a moderate partial correlation (r = .54, controlling for chronological age) between hippocampal and prefrontal composite scores in the DS group, and both composites contributed unique variance to the prediction of MA and adaptive behavior in that group. In sum, these results indicate a particular weakness in hippocampal functions in DS in the context of overall cognitive dysfunction. It is interesting that these results are similar to what has been found in a mouse model of DS. Such a model will make it easier to understand the neurobiological mechanisms that lead to the development of hippocampal dysfunction in DS. PMID- 12625438 TI - Infants' perception of object trajectories. AB - Filling in the gaps in what humans see is a fundamental perceptual skill, but little is known about the developmental origins of occlusion perception. Three experiments were conducted with infants between 2 and 6 months of age to investigate perception of the continuity of an object trajectory that was briefly occluded. The pattern of results across experiments provided little evidence of veridical responses to trajectory occlusion in the youngest infants, but by 6 months, perceptual completion was more robust. Four-month-olds' responses indicated that they perceived continuity under a short duration of occlusion, but when the object was out of sight for a longer interval, they appeared to perceive the trajectory as discontinuous. These results suggest that perceptual completion of a simple object trajectory (and, by logical necessity, veridical object perception) is not functional at birth but emerges across the first several months after onset of visual experience. PMID- 12625439 TI - Life with (or without) father: the benefits of living with two biological parents depend on the father's antisocial behavior. AB - The salutary effects of being raised by two married, biological parents depend on the quality of care parents can provide. Using data from an epidemiological sample of 1,116 5-year-old twin pairs and their parents, this study found that the less time fathers lived with their children, the more conduct problems their children had, but only if the fathers engaged in low levels of antisocial behavior. In contrast, when fathers engaged in high levels of antisocial behavior, the more time they lived with their children, the more conduct problems their children had. Behavioral genetic analyses showed that children who resided with antisocial fathers received a "double whammy" of genetic and environmental risk for conduct problems. Marriage may not be the answer to the problems faced by some children living in single-parent families unless their fathers can become reliable sources of emotional and economic support. PMID- 12625441 TI - U.S. and Korean children's comprehension of fraction names: a reexamination of cross-national differences. AB - Two experiments tested the claim that the transparency of Korean fraction names promotes fraction concepts (Miura, Okamoto, Vlahovic-Stetic, Kim, & Han, 1999). In Experiment 1, U.S. and Korean first and second graders made similar errors on a fraction-identification task, by treating fractions as whole numbers. Contrary to previous findings, Korean children performed at chance when a whole-number representation was included. Nonetheless, Korean children outperformed their U.S. peers overall. In Experiment 2, U.S. children's performance improved when fraction names were used that explicitly referred to part-whole relations like Korean fraction names. U.S. children's scores actually exceeded those of Korean children. Thus, although the differences in fraction names may influence children's performance, this may not account for the reported cross-national differences. PMID- 12625440 TI - Reciprocal influences between stressful life events and adolescent internalizing and externalizing problems. AB - Investigated in this study were hypothesized reciprocal influences between stressful life events and adolescent maladjustment using data from a 6-year, prospective longitudinal study. Stressful life experiences, internalizing symptoms, and externalizing behaviors were assessed for a sample of adolescents (215 males, 236 females) living in the rural Midwest. From 7th to 12th grades, autoregressive analyses showed that stressful life events and these two forms of maladjustment were reciprocally interrelated over time. For example, stressful life events at one point significantly predicted delinquent behaviors 1 year later, which, in turn, significantly predicted stressful life events 1 year later, and vice versa. The findings provide evidence for the accumulating disadvantage for adolescents that results from the mutual reinforcement of problematic situations and adjustment problems over time. PMID- 12625442 TI - Arrows of time in early childhood. AB - Three studies with 149 children were conducted to provide information about development of the perception of temporally unidirectional transformations, such as dropping blocks or breaking a cookie. Children 3.5 through 6.5 years of age compared forward and backward videotapes of events or made individual judgments of what would happen if the actions were attempted. Even children 3.5 to 4.5 years of age recognized the anomaly of backward versions of gravity and separation events. In addition, relatively few children predicted impossible transformations in the prediction task. The results show that young children, like adults, are sensitive to the unidirectional nature of varied transformations. PMID- 12625443 TI - How specific is the shape bias? AB - Children tend to extend object names on the basis of sameness of shape, rather than size, color, or material-a tendency that has been dubbed the "shape bias." Is the shape bias the result of well-learned associations between words and objects? Or does it exist because of a general belief that shape is a good indicator of object category membership? The present three studies addressed this debate by exploring whether the shape bias is specific to naming. In Study 1, 3 year-olds showed the shape bias both when asked to extend a novel name and when asked to select an object of the same kind as a target object. Study 2 found the same shape bias when children were asked to generalize properties relevant to category membership. Study 3 replicated the findings from Study 1 with 2-year olds. These findings suggest that the shape bias derives from children's beliefs about object kinds and is not the product of associative learning. PMID- 12625444 TI - Want that is understood well before say that, think that, and false belief: a test of de Villiers's linguistic determinism on German-speaking children. AB - Two experiments with 79 monolingual German speaking children between 2.5 and 4.5 years showed a consistent developmental gap between children's memory/inference of what someone wanted and what someone wrongly said or thought. For instance, when John is still playing and mother says, "John should be going to bed," more than 70% answered correctly that mother wanted John to go to bed. However, when mother said, "John is going to bed," about 70% answered wrongly that she thought/said that he was still playing. Correct answers emerged with the mastery of the false-belief task. In German, want sentences (about something to happen) obligatorily take the same grammatical that complement as say or think sentences. Therefore, the observed gap constrains de Villier's (1995) linguistic determinism, which claims that acquisition of the necessary grammatical structures for talking about the mind drives children's ability to think about the mind. PMID- 12625445 TI - Parenting and family socialization strategies and children's mental health: low income Mexican-American and Euro-American mothers and children. AB - The extent to which current theories on family-related factors associated with children's depression and conduct problems are applicable to Mexican American children was examined among demographically comparable samples of low-income Mexican American (English and Spanish speaking) and Euro-American mothers and children. There were ethnic differences in mean levels of children's depression, maternal inconsistent discipline, and hostile control. In addition, there were differences across language within the Mexican American sample on levels of reported maternal inconsistent discipline and hostile control. The vast majority of relations between parenting and mental health were similar between Mexican Americans and Euro-Americans, suggesting that current theories do apply across ethnic groups. However, analyses across language within the Mexican American sample showed that language preference moderated the relation between maternal acceptance and children's conduct problems. Moreover, the relation between acceptance and hostile control differed across groups. These results are discussed in light of the relative influence of ethnicity and other contextual variables on parenting and children's mental health. PMID- 12625446 TI - Examination of peer-group contextual effects on aggression during early adolescence. AB - Peer group contextual effects of aggressive behavior among middle school students (6th-8th graders) were examined using a short-term longitudinal design. More specifically, the homophily hypothesis that peer group membership influences individual-level bullying and fighting was evaluated with multilevel sex-specific models of individual- and peer-level aggression scores. Peer groups were identified via social network analysis. Intraclass correlation coefficients yielded through hierarchical linear modeling demonstrated substantial within group similarity on self-reported bullying and fighting, suggesting that students affiliate with individuals who bully and fight at the same frequency. Peer group bullying and fighting was associated with individual-level behavior, even after controlling individual baseline levels for males and females. However, peer contextual effects explained more variance in individual bullying than individual fighting. This differential impact of peer group membership suggests that future studies consider peer relations across subtypes of aggression. PMID- 12625447 TI - The infant as onlooker: learning from emotional reactions observed in a television scenario. AB - Two studies investigated whether 10- and 12-month-olds can use televised emotional reactions to guide their behavior. Infants watched an actress orient toward 1 of 2 novel objects and react with neutral affect during baseline and with positive or negative affect during test. Infants then had 30 s to interact with the objects. In Study 1, 12-month-olds (N = 32) avoided the target object and showed increases in negative affect after observing the negative-emotion scenario. Twelve-month-olds' responses to positive vs. neutral signals did not differ significantly. In Study 2, 10-month-olds (N = 32) attended to the televised presentations but showed no consistent changes in their object interactions or affect. Thus, 12-month-olds used social information presented on television and associated emotional signals with the intended target. PMID- 12625448 TI - Preschool emotional competence: pathway to social competence? AB - Preschoolers' (N = 143) patterns of emotional expressiveness, emotion regulation, and emotion knowledge were assessed. Their contributions to social competence, as evidenced by sociometric likability and teacher ratings, were evaluated via latent variable modeling, both concurrently and across time. Moderation of key results by age and sex was also explored. Emotional competence assessed at 3 to 4 years of age contributed to both concurrent and kindergarten social competence. Even early in the preschool period, contributions of emotional competence to social competence have long-term implications. PMID- 12625449 TI - Anxious solitude and peer exclusion: a diathesis-stress model of internalizing trajectories in childhood. AB - A diathesis-stress model was proposed in which the joint forces of individual vulnerability (anxious solitude) and interpersonal adversity (peer exclusion) predict depressive symptoms in children over time. Children's (N = 388; 50% female) social behavior, peer exclusion, and emotional adjustment were assessed at kindergarten entry and every spring thereafter through 4th grade, primarily by teacher report. Results indicated that anxious solitude and peer exclusion co occur in children soon after kindergarten entry and that anxious solitary children who are excluded early on, in comparison with their nonexcluded anxious solitary counterparts, display greater stability in their subsequent display of anxious solitude. As hypothesized, the joint influence of anxious solitude and exclusion predicted the most elevated depressive symptom trajectories. PMID- 12625450 TI - The moderation of adolescent-to-peer similarity in tobacco and alcohol use by school levels of substance use. AB - Using the National Longitudinal Study of Adolescent Health, this study examined the impact of school-level smoking and drinking on adolescent-peer similarity for smoking and drinking. Hierarchical linear modeling revealed that adolescent-peer similarity was significantly moderated by school-level substance use for both tobacco and alcohol use. For tobacco, similarity between adolescent and peer use increased from .18 in the lower quartile of tobacco-using schools to .44 in the upper quartile of tobacco-using schools. Corresponding similarities for alcohol use ranged from .25 to .34. These results suggest that schools with relatively few substance-using peers provide less opportunities for adolescents to pick niches that expose them to risk factors that correspond to their own substance use behaviors. PMID- 12625451 TI - A secure base in adolescence: markers of attachment security in the mother adolescent relationship. AB - This study sought to identify ways in which adolescent attachment security, as assessed via the Adult Attachment Interview, is manifest in qualities of the secure base provided by the mother-adolescent relationship. Assessments included data coded from mother-adolescent interactions, test-based data, and adolescent self-reports obtained from an ethnically and socioeconomically diverse sample of moderately at-risk 9th and 10th graders. This study found several robust markers of adolescent attachment security in the mother-adolescent relationship. Each of these markers was found to contribute unique variance to explaining adolescent security, and in combination, they accounted for as much as 40% of the raw variance in adolescent security. These findings suggest that security is closely connected to the workings of the mother-adolescent relationship via a secure-base phenomenon, in which the teen can explore independence in thought and speech from the secure base of a maternal relationship characterized by maternal attunement to the adolescent and maternal supportiveness. PMID- 12625452 TI - Preschool children use linguistic form class and pragmatic cues to interpret generics. AB - Generic noun phrases (e.g., "Bats live in caves") are important for expressing knowledge about abstract kinds. Past work has found that parents frequently use generic noun phrases in their speech to young children. However, little is known regarding how children understand these expressions, nor which cues signal generic meaning. The present set of 5 studies examined the influence of linguistic form class (e.g., "What color are dogs?" [generic] versus "What color are the dogs?" [nongeneric]) and of pragmatic context (e.g., "What color are they?" in the presence of either a single exemplar [generic] or multiple exemplars [nongeneric]). Participants were 2-year-olds (N = 42), 3-year-olds (N = 40), 4-year-olds (N = 40), and adults (N = 51). The data indicate that by 2 years of age, children use linguistic form class, and by 3 years of age, children use pragmatic context. These findings demonstrate that young children have begun to understand the distinction between generic and nongeneric noun phrases from a very young age, and that identification of generics is signaled not by formal or pragmatic cues alone, but by a combination of information from both language form and pragmatic context. It is suggested that children make use of multiple linguistic and conceptual cues to acquire and interpret generics. PMID- 12625454 TI - Distribution and morphology of serotonin-immunoreactive axons in the hippocampal region of the New Zealand white rabbit. I. Area dentata and hippocampus. AB - This study provides a detailed light microscopic description of the morphology and distribution of immunohistochemically stained serotonergic axons in the hippocampal region of the New Zealand white rabbit. The serotonergic axons were segregated morphologically into three types: beaded fibers, fine fibers, and stem axons, respectively. Beaded fibers were thin serotonergic axons with large varicosities, whereas thin axons with small fusiform or granular varicosities were called fine fibers. Finally, thick straight non-varicose axons were called stem-axons. Beaded fibers often formed large conglomerates with numerous boutons (pericellular arrays) in close apposition to the cell-rich layers in the hippocampal region, e.g., the granular and hilar cell layers of the dentate area and the pyramidal cell layer ventrally in CA3. The pericellular arrays in these layers were often encountered in relation to small calbindin-D2BK-positive cells, as shown by immunohistochemical double staining for serotonin and calbindin-D28K. The beaded and fine serotonergic fibers displayed a specific innervation pattern in the hippocampal region and were encountered predominantly within the terminal field of the perforant path, e.g., the stratum moleculare hippocampi and the outer two-thirds of the dentate molecular layer. These fibers were also frequently seen in the deep part of the stratum oriens and the alveus, forming a dense plexus in relation to large multipolar calbindin-D28K-positive cells and their basal extensions. Stem-axons were primarily seen in the fimbria and alveus. This innervation pattern was present throughout the entire hippocampal formation, but there were considerable septotemporal differences in the density of the serotonergic innervation. A high density of innervation prevailed in the ventral/temporal part of the hippocampal formation, whereas the dorsal/septal part received only a moderate to weak serotonergic innervation. These results suggest that the serotonergic system could modulate the internal hippocampal circuitry by way of its innervation in the terminal field of the perforant path, the hilus fasciae dentatae, and ventrally in the zone closely apposed to the mossy fiber layer and the pyramidal cells of CA3. This modulation could be of a dual nature, mediated directly by single serotonergic fibers traversing the hippocampal layers or indirectly by the pericellular arrays and their close relation to the calbindin-D28K-positive cells. The marked septotemporal differences in innervation density point toward a difference between the ventral and dorsal parts of the hippocampal formation with respect to serotonergic function and need for serotonergic modulation. PMID- 12625453 TI - Temporal specific patterns of semaphorin gene expression in rat brain after kainic acid-induced status epilepticus. AB - Mossy fiber sprouting and other forms of synaptic reorganization may form the basis for a recurrent excitatory network in epileptic foci. Four major classes of axon guidance molecules--the ephrins, netrins, slits, and semaphorins--provide targeting information to outgrowing axons along predetermined pathways during development. These molecules may also play a role in synaptic reorganization in the adult brain and thereby promote epileptogenesis. We studied semaphorin gene expression, as assessed by in situ hybridization, using riboprobes generated from rat cDNA in an adult model of synaptic reorganization, kainic acid (KA)-induced status epilepticus (SE). Within the first week after KA-induced SE, semaphorin 3C, a class III semaphorin, mRNA content is decreased in the CA1 area of the hippocampus and is increased in the upper layers of cerebral cortex. Another class III semaphorin, semaphorin 3F, is also decreased in CA1 and CA3 of hippocampus within the first week after KA-SE. These changes in gene expression are principally confined to neurons. By contrast, there was little change in the semaphorin 4C mRNA content of CA1 neurons at this time. No changes in expression of semaphorin 3A and 4C genes were detected 28 days after KA-induced SE. Regulation of semaphorin gene expression after KA-induced SE suggests that neurons may regulate the expression of axonal guidance molecules and thereby contribute to synaptic reorganization after injury of the mature brain. The anatomic locale of the altered semaphorin gene expression may serve as a marker for specific networks undergoing synaptic reorganization in the epileptic brain. PMID- 12625455 TI - Effect of halothane on type 2 immobility-related hippocampal theta field activity and theta-on/theta-off cell discharges. AB - Rats were studied in acute and chronic (freely moving) recording conditions during exposure to different levels of the volatile anesthetic halothane, in order to assess effects on hippocampal theta field activity in the chronic condition and on theta-related cellular discharges in the acute condition. Previous work has shown that the generation of hippocampal type 2 theta depends on the coactivation of cholinergic and GABAergic inputs from the medial septum. Based on these data and recent findings that halothane acts on interneuron GABA(A) receptors, we predicted that exposure of rats to subanesthetic levels would result in the induction of type 2 theta field activity. In the chronic condition, exposure to subanesthetic levels of halothane (0.5-1.0 vol %) was found to induce theta field activity during periods of immobility (type 2 theta) with a mean increase of 39% in amplitude (mV) compared to control levels during movement. The total percentage of signal power (V2) associated with peak theta frequencies (80% compared to control levels of 47%) was also increased by halothane. Over the whole range of administered halothane concentrations, theta field frequency progressively declined from a mean peak frequency of 6.5 +/- 0.8 Hz at 0.5 vol % halothane to a mean peak frequency of 4.0 +/- 1.8 Hz at 2.0 vol % halothane. Subsequent administration of a muscarinic cholinergic antagonist, atropine sulfate, selectively abolished all type 2 immobility-related theta field activity, while type 1 movement-related theta was still intact. At anesthetic levels (1.5-2.0 vol %) in acute experiments, hippocampal field activity spontaneously cycled between theta and large-amplitude irregular activity. Analysis of depth profiles in four experiments revealed they were identical to those previously described for rats under urethane anesthesia conditions. In addition, the discharge properties of 31 theta-related cells, classified as tonic and phasic theta-on and tonic and phasic theta-off cells, did not differ significantly from those described previously in rats anesthetized with urethane. These data provide further support for an involvement of GABA(A) receptors in the generation of hippocampal theta. PMID- 12625456 TI - Selective impairment of fornix-transected rats on a new nonspatial, odor-guided task. AB - In the present experiment, sham-operated (SH) and fornix-transected (FX) rats were trained on a new nonspatial, odor-guided task. On each session, eight odor pairs were presented twice. On the first occurrence of a pair, rats were reinforced for pushing the container (go response) in which the olfactory stimuli were placed. On the second occurrence, they were not reinforced and had to refrain from responding (no-go response) to be scored as success. Rats were first trained to criterion on odor pairs made of replicates of the same odor (S pairs). Then they were trained to criterion on pairs made of different odors, each member of the pair overlapping with that of another pair (O pairs) and finally, on pairs of different odors with no overlap (NO pairs). The results showed that the number of sessions to reach criterion was significantly higher in FX than in SH rats during training on O pairs, but not during training on S or on NO pairs. These findings are consistent with the configural (Rudy and Sutherland, 1995: Hippocampus 5:375-389) or relational (Eichenbaum et al., 1994: Behav Brain Sci 17:449-518) account of the hippocampal memory function. PMID- 12625457 TI - The role of NMDA glutamate receptors, PKA, MAPK, and CAMKII in the hippocampus in extinction of conditioned fear. AB - Pavlovian conditioning involves the association of initially neutral conditioned stimuli (CS) with unconditioned stimuli (US) that elicit a response. In contextual fear conditioning in rodents, the CS is the context of a training apparatus and the US is a foot shock. Retrieval of memory of the training is tested by presenting the CS alone. But a retrieval test also initiates extinction of the conditioned response. That is, presentation of the CS alone results in new learning, i.e., the CS no longer predicts the US. Here we report that extinction is triggered by two hippocampal signaling pathways underlying retrieval (the cAMP dependent protein kinase and the mitogen-activated protein kinase pathways) and two other mechanisms that become activated at the same time and are not necessary for retrieval (N-methyl-D-aspartate glutamatergic receptors and the calcium/calmodulin-dependent protein kinase II signaling pathway). Thus, the molecular mechanisms underlying acquisition and/or consolidation of the memory for extinction are similar to those described for the acquisition and/or consolidation of the original contextual fear. PMID- 12625458 TI - Type II glucocorticoid receptor immunoreactivity in the mossy cells of the rat and the mouse hippocampus. AB - Hippocampal principal neurons, granule and pyramidal cells, are known to express type II glucocorticoid receptors (GR) and it is believed that glucocorticoids (GC) mediate at least some of their effects through GR. Under conditions of severe stress and trauma, these principal cells are vulnerable to damage and this mechanism may be exacerbated by GR. The mossy cell, an excitatory dentate gyrus neuron, is also damaged following trauma, with over 50% reported loss in rats after kainate-induced seizures. However, it has not been determined if GC play any role in protecting or exacerbating damage to this important hippocampal cell type. In the present study, we have undertaken an evaluation of the presence of GR in mossy cells of the rat and mouse utilizing an immunocytochemical double labeling technique. To identify mossy cells in the rat, we utilized an antibody to the glutamate receptor subunit 2/3 (GluR2/3). In addition to GluR2/3 antibodies, in the mouse, an antibody to the calcium-binding protein, calretinin (CR), to identify mossy cells was also employed. Our results show that GR immunoreactivity (IR) was colocalized with GluR2/3-IR in approximately 90% of the rat and the mouse mossy cells. In addition, GR-IR was identified in the CR-IR mossy cells in the mouse hippocampus, whereas the CR-IR interneurons of rat and mouse were negative for GR-IR. The presence of GR on mossy cells may indicate the ability of GC to mediate cellular activity of these cells. PMID- 12625459 TI - Encoding of novel picture pairs activates the perirhinal cortex: an fMRI study. AB - It is well established in nonhuman primates that the medial temporal lobe (MTL) structures, the hippocampus and the entorhinal and perirhinal cortices, are necessary for declarative memory encoding. In humans, the neuropathological and neuropsychological changes in early Alzheimer's disease (AD) further support a role for the rhinal cortex in the consolidation of new events into long-term memory. Little is known, however, regarding the function of the rhinal cortex in humans in vivo. To examine the participation of the interconnected MTL structures as well as the whole-brain network of activated brain areas in visual associative long-term memory, functional magnetic resonance imaging (fMRI) was used to determine the brain regions that are activated during encoding and retrieval of paired pictures in 12 young control subjects. The most striking finding in the MTL activation pattern was the consistent activation of the perirhinal cortex in the encoding-baseline and encoding-retrieval comparisons with a strict statistical threshold (P < 0.00001). In contrast, no perirhinal cortex activation was detected in the retrieval-baseline or retrieval-encoding comparisons even with a low statistical threshold (P < 0.05). The location of the perirhinal activation area was in the transentorhinal part of the perirhinal cortex, in the medial bank of the collateral sulcus. The hippocampus and the more posterior parahippocampal gyrus were activated in both encoding and retrieval conditions. During the encoding processing, MTL activations were more consistent and the hippocampal activation area located more anteriorly than during retrieval. The frontal, parietal, temporal, and occipital association cortices were also activated in the encoding-baseline and retrieval-baseline comparisons. The data suggest that encoding, but not retrieval, of novel picture pairs activates the perirhinal cortex. To our knowledge, this is the first fMRI study reporting encoding activation in this transentorhinal part of the perirhinal cortex, the site of the very earliest neuropathological changes in AD. PMID- 12625460 TI - Long-term potentiation and long-term depression induced by local application of ATP to hippocampal CA1 neurons of the guinea pig. AB - The present study has investigated the role of ATP in the induction of synaptic plasticity, using local application of ATP by picopump administration into the stratum radiatum of guinea pig hippocampal region CA1. Excitatory postsynaptic currents (EPSCs) evoked by stimulation of Schaffer collateral/commissural afferents synapsing on CA1 pyramidal cells of hippocampal slices were monitored in voltage-clamp mode, using whole-cell recording. Brief local application of ATP (1 mM) induced an inward current, usually consisting of early- and late-phase components. Because the late-phase component of an ATP-induced current was largely inhibited by Ca2+-free solution, this component is supposed to depend on extracellular Ca2+. After local application of ATP, long-term synaptic modification of EPSCs was induced: LTP was detected in neurons exhibiting a small late Ca2+ current, while LTD was obtained from recordings showing a large late Ca2+ current in response to ATP application. There was a statistically significant correlation between the magnitude of long-term plastic changes and the size of Ca2+ currents in response to ATP application. Furthermore, there was significant difference between the average size of the Ca2+ current in the LTP group and the size in the LTD group. These results suggest that a small Ca2+ influx in response to ATP application induces LTP, whereas a large one induces LTD in guinea pig hippocampal CA1 neurons. PMID- 12625461 TI - Theta- and movement velocity-related firing of hippocampal neurons is disrupted by lesions centered on the perirhinal cortex. AB - The hippocampus is critically involved in spatial memory and navigation. It has previously been proposed that, as part of this process, the hippocampus might have access to self-motion information. The possibility that some of this information may originate from the perirhinal cortex, a region involved in high order multimodal processing, was tested in the present study by recording the responses of hippocampal complex-spike (place cells) and theta cells (putative interneurons) to movement velocity and to the movement-related theta rhythm EEG while rats with bilateral ibotenic acid lesions centered on the perirhinal cortex (n = 5), or control surgeries (n = 5), foraged in a rectangular environment. Perirhinal cortex lesions altered several characteristics of place and theta cell firing. First, the proportion of theta cells recorded was significantly lower in perirhinal lesion animals (8/39 units) compared to controls (22/53 units). Second, the firing of place cells recorded from lesion animals was phase-shifted so as to occur significantly earlier during the theta rhythm cycle than in place cells from controls (mean difference = 48.73 degrees). Third, the firing rates of a significantly lower proportion of place cells from lesion animals were modulated by the movement velocity of the animal compared to place cells from controls. These results indicate that the perirhinal cortex contributes to the responses of hippocampal CA1 place cells by providing information about self movement and by controlling the timing of firing of these cells. This information may normally be utilized by the hippocampus during spatial memory and navigation processes. PMID- 12625462 TI - Stimulation-induced reset of hippocampal theta in the freely performing rat. AB - Previous research has suggested that visual and auditory stimuli in a working memory task have the ability to reset hippocampal theta, perhaps allowing an organism to encode the incoming information optimally. The present study examined two possible neural pathways involved in theta resetting. Rats were trained on a visual discrimination task in an operant chamber. At the beginning of a trial, a light appeared over a centrally located lever that the rat was required to press to receive a water reward. There was a 30-s intertrial interval before the next light stimulus appeared. After learning the task, all rats received surgical implantation of stimulating electrodes in both the fornix and the perforant path and recording electrodes, bilaterally in the hippocampus. After surgery, theta was recorded before and after the light stimulus to determine whether resetting to the visual stimulus occurred. During the intertrial interval, rats received single-pulse electrical stimulation of either the fornix or perforant path. Theta was recorded both before and after the electrical stimulation to determine whether resetting occurred. In this experiment, hippocampal theta was reset after all three stimulus conditions (light, perforant path, and fornix stimulation), with the greatest degree of reset occurring after the fornix stimulation. The results suggest that activation of the perforant path and fornix may underlie theta reset and provide a mechanism by which the hippocampus may enhance cognitive processing. PMID- 12625463 TI - Reward value invariant place responses and reward site associated activity in hippocampal neurons of behaving rats. AB - To investigate the involvement of the hippocampal-accumbens system in goal oriented displacement behaviors, hippocampal neuronal activity was recorded in rats learning and recalling new distributions of different volumes of liquid reward among the arms of a plus maze. Each arm had a reward box containing a water trough and identical visual cues that could be illuminated independently. As the water-restricted rat successively visited the respective boxes, it received 7, 5, and 3 drops of water, and then 1 drop, provided at 1-s intervals. (Reward distributions were reassigned daily and mid-session.) In the training phase, reward boxes were lit individually. In the recall phase, the lamps on all arms were lit and then turned off as the rat visited the boxes in order of descending value. Neuronal firing rates were analyzed for changes related to reward value or to shifts between learning and recall phases. The principal finding is that place responses remained unchanged after these manipulations and that these neurons showed no evidence of explicit coding of reward value. In addition, two other types of responses appeared while the rat was stationary at the reward boxes awaiting multiple rewards. These were observed primarily in neurons within the dentate gyrus, but also in CA1. Position-selective reward site responses were regular at 20-60 impulses per second, while position-independent discharges bursted irregularly at about 5 impulses per second. Such responses could explain controversial reports of reward dependence in hippocampal neurons. The higher incidence of the latter responses in the temporal ("ventral") hippocampus is consistent with the distinctive anatomical and functional properties of this subregion. PMID- 12625464 TI - The entorhinal cortex of the mouse: organization of the projection to the hippocampal formation. AB - The origin and the terminations of the projections from the entorhinal cortex to the hippocampal formation of the mouse (C57BL/6J strain) have been studied using anterogradely and retrogradely transported tracers. The entorhinal cortex is principally divided into two areas, the lateral entorhinal area (LEA) and the medial entorhinal area (MEA). LEA is the origin of the lateral perforant path that terminates in the outer one-third of the molecular layer of the dentate gyrus, and MEA is the origin of the medial perforant path that ends in the middle one-third of the molecular layer of the dentate gyrus. This projection is mostly to the ispsilateral dentate gyrus; only a few labeled axons and terminals are found in the contralateral dentate gyrus. The projection to the dentate gyrus originates predominantly from neurons in layer II of the entorhinal cortex. The entorhinal cortex also projects to CA3 and CA1 and to subiculum; in both CA3 and CA1, the terminals are present in stratum lacunosum-moleculare, whereas in the subiculum the terminals are in the outer part of the molecular layer. The projection from the entorhinal cortex to CA3, CA1, and subiculum is bilateral, and it originates predominantly from neurons in layer III, but a small number of neurons in the deeper layers of the entorhinal cortex contributes to this projection. The projection of entorhinal cortex to the hippocampus is topographically organized, neurons in the lateral part of both LEA and MEA project to the dorsal part (i.e., septal pole) of the hippocampus, whereas the projection to the ventral (i.e., temporal pole) hippocampus originates from neurons in medial parts of the entorhinal cortex. PMID- 12625465 TI - Cellular mechanisms underlying the rhythmic bursts induced by NMDA microiontophoresis at the apical dendrites of CA1 pyramidal neurons. AB - This article reports the cellular mechanisms underlying a form of intracellular "theta-like" (theta-like) rhythm evoked in vitro by microiontophoresis of N methyl-D-aspartate (NMDA) at the apical dendrites of CA1 pyramidal neurons. Rhythmic membrane potential (Vm) oscillations and action potential (AP) bursts (approximately 6 Hz; approximately 20 mV; approximately 2-5 APs) were evoked in all cells. The response lasted approximately 2 s, and the initial oscillations were usually small (< 20 mV) and below AP threshold. Rhythmic bursts were never evoked by imposed depolarization in the absence of NMDA. Block of Na+ conductance with tetrodotoxin (TTX) (1.5 microM), of non-NMDA receptors with 6-cyano-7 nitroquinoxaline-2,3-dione (CNQX) (20 microM) and of synaptic inhibition by bicuculline (50 microM) and picrotoxin (50 microM) did not prevent NMDA oscillation. Inhibition of the voltage dependence of the NMDA conductance in Mg2+ free Ringer's solution blocked oscillations. Preventing Ca2+ influx with Ca2+ free and Co2+ (2-mM) solutions and block of the slow Ca2+-dependent afterhyperpolarization (sAHP) by carbamilcholine (5 microM), isoproterenol (10 microM), and intracellular BAPTA blocked NMDA oscillations. Inhibition of L-type Ca2+ conductance with nifedipine (30 microM) reduced oscillation amplitude. Block of tetraethylammonium (TEA) (10 mM) and 4AP (10 mM)-sensitive K+ conductance increased the duration and amplitude, but not the frequency, of oscillations. In conclusion, theta-like bursts relied on the voltage dependence of the NMDA conductance and on high-threshold Ca2+ spikes to initiate and boost the depolarizing phase of oscillations. The repolarization is initiated by TEA sensitive K+ conductance and is controlled by the sAHP. These results suggest a role of interactions between NMDA conductance and intrinsic membrane properties in generating the CA1 theta-rhythm. PMID- 12625466 TI - Regional analysis of hippocampal activation during memory encoding and retrieval: fMRI study. AB - Investigators have recently begun to examine the differential role of subregions of the hippocampus in episodic memory. Two distinct models have gained prominence in the field. One model, outlined by Moser and Moser (Hippocampus 1998;8:608 619), based mainly on animal studies, has proposed that episodic memory is subserved by the posterior two-thirds of the hippocampus alone. A second model, derived by Lepage et al. (Hippocampus 1998;8:313-322) from their review of 52 PET studies, has suggested that the anterior hippocampus is activated by memory encoding while the posterior hippocampus is activated by memory retrieval. Functional magnetic resonance imaging (fMRI) studies have tended to show limited activation in the anteriormost regions of the hippocampus, providing support for the Moser and Moser model. A potential confounding factor in these fMRI studies, however, is that susceptibility artifact may differentially reduce signal in the anterior versus the posterior hippocampus. In the present study, we examined activation differences between hippocampal subregions during encoding and retrieval of words and interpreted our findings within the context of these two models. We also examined the extent to which susceptibility artifact affects the analysis and interpretation of hippocampal activation by demonstrating its differential effect on the anterior versus the posterior hippocampus. Both voxel by-voxel and region-of-interest analyses were conducted, allowing us to quantify differences between the anterior and posterior aspects of the hippocampus. We detected significant hippocampal activation in both the encoding and retrieval conditions. Our data do not provide evidence for regional anatomic differences in activation between encoding and retrieval. The data do suggest that, even after accounting for susceptibility artifact, both encoding and retrieval of verbal stimuli activate the middle and posterior hippocampus more strongly than the anterior hippocampus. Finally, this study is the first to quantify the effects of susceptibility-induced signal loss on hippocampal activation and suggests that this artifact has significantly biased the interpretation of earlier fMRI studies. PMID- 12625467 TI - Role of neuron-glia interaction in the regulation of brain glucose utilization. PMID- 12625468 TI - Glucosensing neurons: the metabolic sensors of the brain? PMID- 12625469 TI - Glucose transport into brain: effects of hypoglycemia. PMID- 12625470 TI - Brain glycogen: an insulin-sensitive carbohydrate store. PMID- 12625472 TI - Brain glucose metabolism and hypoglycaemia. PMID- 12625471 TI - Hypoglycemia and white matter: pathophysiology of axon injury and role of glycogen. PMID- 12625473 TI - Hepatorenal glucose reciprocity in physiologic and pathologic conditions. PMID- 12625474 TI - The role of neural signalling and hypoglycaemia per se in the counterregulatory response. PMID- 12625475 TI - Portal vein glucose sensing. PMID- 12625476 TI - Portal vs central glucose sensing in humans: the value of clinical models. PMID- 12625477 TI - The anatomy of the human hypoglycaemia sensor. PMID- 12625478 TI - Autonomic mechanism and defects in the glucagon response to insulin-induced hypoglycaemia. AB - In summary, this article briefly reviews the evidence that three separate autonomic inputs to the islet are capable of stimulating glucagon secretion and that each is activated during IIH. We have reviewed our evidence that these autonomic inputs mediate the glucagon response to IIH, both in non-diabetic animals and humans. Finally, we outline our new preliminary data suggesting an eSIN in an autoimmune animal model of T1DM. We conclude that the glucagon response to IIH is autonomically mediated in non-diabetic animals and humans. We further suggest that at least one of these autonomic inputs, the sympathetic innervation of the islet, is diminished in autoimmune T1DM. These data raise the novel possibility that an autonomic defect contributes to the loss of the glucagon response to IIH in T1DM. PMID- 12625479 TI - Adrenaline vs glucagon in the primacy of glucose counterregulation. PMID- 12625480 TI - The pathophysiology of hypoglycaemia in diabetes. PMID- 12625482 TI - Reciprocity of hypoglycaemia and exercise in blunting respective counterregulatory responses: possible role of cortisol as a mediator. PMID- 12625481 TI - Alterations in brain glucose uptake and hypoglycaemia unawareness. PMID- 12625483 TI - Mechanisms of impaired hypothalamic-pituitary-adrenal (HPA) function in diabetes: reduced counterregulatory responsiveness to hypoglycaemia. AB - In summary, our data suggest that in uncontrolled diabetes, increased HPA activity is caused by increased central drive at or above the level of the PVN. Insulin treatment only restores HPA activity at and below the pituitary level, presumably by GC-mediated suppression of ACTH secretion. We hypothesize that the defective HPA response to hypoglycaemia is at least in part due to a lack of a decrease in MR mRNA in response to hypoglycaemia, and diminished sensitivity of the pituitary and adrenal gland to stimulation. Interestingly, insulin treatment restores the HPA response, but not the defective epinephrine response. Therefore, defective epinephrine responses are not linked to defective HPA responses. Similarly, antecedent hypoglycaemia specifically impairs epinephrine responses, but not HPA responses to hypoglycaemia. These studies have revealed some of the mechanisms of impaired HPA function in diabetes and its impaired responsiveness to hypoglycaemia. Further investigations are essential for understanding poor counterregulation in insulin-treated diabetes and may lead to new strategies for preventing hypoglycaemia. PMID- 12625484 TI - The role of beta-adrenergic sensitivity in the pathogenesis of hypoglycaemia unawareness. PMID- 12625485 TI - Cognitive function in acute hypoglycaemia 2. PMID- 12625486 TI - Cognitive function in hypoglycaemia. PMID- 12625487 TI - What we know about counterregulation in Type 2 diabetes. PMID- 12625488 TI - The effects of hormonal therapy on aortic stiffness and left ventricular diastolic function. AB - OBJECTIVE: The main objective of this study is to investigate the effects of oestrogen replacement therapy (ERT) and hormone replacement therapy (HRT) on aortic stiffness and on the left ventricular diastolic function, including tissue Doppler. METHODS AND RESULTS: The two study groups were composed of 20 postmenopausal women having HRT and 22 postmenopausal women having ERT. Each group was evaluated for aortic elasticity properties and the left ventricular diastolic function at both the pre-treatment stage and after 12 weeks of hormonal therapy. There was a significant improvement in beta index (5.2+/- 2.5 vs. 3.2+/- 2.2, p = 0.001), distensibility (5.2+/-3.7 vs. 6.1 +/-4.1 cm2 x dyn(-1) x 10(-3), p = 0.036) and mitral E/Em ratio (7.44 +/- 3.25 vs. 5.75 +/- .2.34, p = 0.004) with ERT. HRT was observed to improve aortic elasticity properties (for strain 10.7+/-4.7 vs. 12.8 +/-7.6%, for beta index 4.9+/-2.1 vs. 3.39+/- 2.4 and for distensibility 4.6+/- 2.1 vs.5.69 +/-4.1 cm2 x dyn(-1) x 10(-3)) and the mitral E/Em ratio (7.61 +/- 3.31 vs.5.81 +/-2.31, p = 0.003). CONCLUSION: Both ERT and HRT have an improving effect on aortic elasticity properties, as well as on the diastolic function. PMID- 12625489 TI - Hyperlipidaemias and serum cytokines in patients with coronary artery disease. AB - OBJECTIVE: The inflammatory processes as well as the lipid disturbances play an important role in the pathogenesis of atherosclerosis. The aim of the study was to evaluate the influence of the hyperlipidaemias on serum levels of tumour necrosis factor (TNF) alpha, the soluble form of TNF receptor (sTNFR) 1 and 2, Interleukin (IL)-10 in patients with stable coronary artery disease (CAD). METHODS AND RESULTS: The study group comprised 94 consecutive admissions with stable CAD: 39 patients with hypercholesterolaemia (group HC), 22 patients with mixed hyperlipidaemia (group HL) and 33 patients with normal lipids (group NL). Twenty healthy volunteers were the controls (group C). Serum TNFalpha levels were higher in all CAD groups (p < 0.001) than in healthy subjects. Mean serum concentrations of sTNFR 1 were significantly higher in group NL (p < 0.05) in comparison both to group HC and controls. IL-10 levels were higher in group HC than in controls (p < 0.5). In all CAD patients TNFalpha showed a negative correlation with HDL-cholesterol (p < 0.001) and a positive correlation with triglycerides (p < 0.00 1). Moreover, sTNFR 1 and IL-10 showed a negative (p < 0.05) and sTNFR 2 a positive correlation with LDL-cholesterol (p < 0.001). CONCLUSIONS: CAD patients are characterized by increased serum concentrations of TNFalpha. It seems likely that immune activation (TNFalpha, sTNFR 1, sTNFR 2, and IL- 10) in CAD patients is related to serum lipids levels. PMID- 12625490 TI - Chest pain after partial upper versus complete sternotomy for aortic valve surgery. AB - OBJECTIVE: The aim of this study was to evaluate pain and pulmonary function after partial upper versus complete sternotomy for aortic valve surgery. METHODS AND RESULTS: In a prospective study of 31 adult patients, 15 individuals underwent aortic valve replacement via partial upper sternotomy, and 16 patients were operated on by means of a full median sternotomy. Chest pain was assessed at rest, after spirometry and coughing, using the visual analogue scale and a structured questionnaire. Forced expiratory volume at I second and forced vital capacity were measured by a hand-held spirometer. Minimal-access patients had less postoperative pain and improved pulmonary function in comparison with conventional aortic valve replacement. After partial upper sternotomy, the postoperative chest pain was confined to the sternal area, while after standard sternotomy a more generalized chest pain occurred. Differences in pain scores could still be demonstrated one month after discharge. CONCLUSIONS: This study suggests that partial upper sternotomy improves pulmonary function and reduces pain in comparison with standard full sternotomy. PMID- 12625492 TI - Pacemaker-related endocarditis: the value of transoesophageal echocardiography in diagnosis and treatment. AB - We report 5 cases with pacemaker endocarditis. All cases were presented with long standing fever and the diagnosis was confirmed by transoesophageal echocardiography. Besides the contribution to diagnosis, transoesophageal echocardiography defined the technique that was applied to remove the pacing system. In four patients with large vegetations (> 10 mm) the entire pacing system was removed by thoracotomy, while in one with relatively small vegetations (< or = 10 mm) the pacing system was removed by external traction. We have reviewed the literature regarding pacemaker endocarditis, particularly with respect to the role of transoesophageal echocardiography in the early diagnosis and determination of optimal treatment. PMID- 12625491 TI - Is coronary angiography performed in the appropriate patients after acute myocardial infarction? Clinical and angiographic findings and one-year mortality in a large unselected cohort of 2493 consecutive patients in a French region (the PRIMA study). AB - OBJECTIVE: Rates of coronary angiography (CA) after myocardial infarction (MI) vary widely between institutions. Furthermore, the indications for CA are often in conflict with recognized guidelines. The present study sought to determine the characteristics and the one-year mortality in patients with MI, regardless of age and hospital facilities, according to the use of CA after MI. METHODS AND RESULTS: Data were prospectively collected in all patients with MI admitted to all hospitals in three departments in the Rhone-Alpes region. Among 2493 patients, 1117 (45%) underwent CA. In multivariate analysis, CA rate was lower with increasing age, female sex, in patients with comorbidity or heart failure. CA was performed in 49% of patients admitted to hospitals with on-site CA vs. 32% in hospitals without on-site CA (OR: 3.54, after adjustment for patients' characteristics). One-year mortality rate was 6.5% for the CA group and 36.9% for the no-CA group. In multivariate analysis, age, history of angina pectoris, presence of Q waves, Killip class at admission II, III, or IV and CPK ratio > or = 9 were significant predictors of a higher one-year mortality, but performance of CA did not significantly influence it: RR: 0.79 (95% CI 0.58 to 1.07). CONCLUSIONS: Among patients with MI in a large unselected cohort in a French region, the one-year mortality was significantly lower in those referred for angiography. However, after correction for the confounding effects of simple baseline clinical indicators of risk, this apparent benefit reflected the fact that angiography was performed in those at lowest risk. PMID- 12625494 TI - Cor triatriatum dextrum. Surgical treatment in a neonate. AB - This is the report of a cor triatriatum dextrum, detected in utero and surgically treated at 10 days of age. We have been unable to find a similar report of neonatal correction in the literature. PMID- 12625493 TI - Myocardial infarction and primary thrombocytosis in a young man. AB - We report the case of a 27-year-old male smoker with simultaneous myocardial infarction (MI) and acute peripheral ischaemia. At the time of his first hospitalization, the platelet count and the coronary angiogram were normal. Two years later, the patient presented again with a leg artery occlusion, followed, a few days later, by a recurrence of MI. At this time, the platelet count was high and the coronary angiogram revealed an occlusion of the right coronary artery. The diagnosis of primary thrombocytosis was made by bone marrow aspiration and biopsy. Different unusual mechanisms of MI in young people are discussed in this report. PMID- 12625495 TI - Thromboprophylaxis in general medical patients. PMID- 12625496 TI - Markers of inflammation in atrial fibrillation. PMID- 12625497 TI - Pregnancy in perinatally HIV-infected adolescents and young adults--Puerto Rico, 2002. AB - Since the introduction of highly active antiretroviral (ARV) therapy in the United States in the mid-1990s, the life expectancy of U.S. children who were infected perinatally with human immunodeficiency virus (HIV) has increased substantially. As a result, the number of perinatally HIV-infected females in the United States who are becoming both sexually active and pregnant is increasing. During August 1998-May 2002, a total of 10 pregnancies were identified among eight perinatally HIV-infected adolescents and young adults in Puerto Rico; in April 2002, the Puerto Rico Department of Health (PRDOH) asked CDC to assist in assessing such pregnancies. This report describes these pregnancies and discusses factors associated with sexual activity and pregnancy. The findings suggest that increasing numbers of pregnancies will occur among perinatally HIV-infected adolescents and young adults and that appropriately tailored reproductive health interventions should be developed. PMID- 12625498 TI - Secondhand smoke exposure among middle and high school students--Texas, 2001. AB - Secondhand smoke (SHS) exposure causes approximately 3,000 deaths each year in Texas and is associated with increased morbidity caused by acute respiratory infections and asthma among children and respiratory problems among adults. Adult lung cancer risk might be increased as a result of SHS exposure during childhood and adolescence. To assess SHS exposure among middle and high school students in Texas, the Texas Department of Health (TDH) analyzed data from the 2001 Texas Youth Tobacco Survey (TYTS). This report summarizes the results of that analysis, which indicate that half of 8,687 middle school students and two-thirds of 8,696 high school students reported SHS exposure within the 7 days preceding the survey. The prevalence of SHS exposure was higher among students who reported living with a cigarette smoker or having at least one close friend who smokes. Among middle school students, the prevalence of SHS exposure was lower among those who perceived SHS to be harmful to their health. These findings emphasize the need for continuing efforts to decrease SHS exposure among youth and to increase understanding of the harmful health effects of SHS exposure. PMID- 12625499 TI - Ambulance crash-related injuries among Emergency Medical Services workers--United States, 1991-2002. AB - Ambulance crashes are one of many hazards faced by Emergency Medical Services (EMS) personnel. Although no complete national count of ground ambulance crashes exists, the total number of fatal crashes involving ambulances can be ascertained by using the National Highway Traffic Safety Administration (NHTSA) Fatality Analysis Reporting System (FARS). To characterize risk factors for EMS workers involved in ambulance crashes, CDC's National Institute for Occupational Safety and Health (NIOSH) and NHTSA investigated three case reports of ambulance crashes. This report summarizes these investigations, presents surveillance data, and discusses recommendations for prevention measures. NIOSH is identifying and testing alternative measures to reduce injury risk for EMS workers. PMID- 12625500 TI - Smallpox vaccine adverse events among civilians--United States, February 18-24, 2003. AB - During the civilian smallpox vaccination program, CDC and state health departments are conducting surveillance for vaccine-associated adverse events. In the first stage of the program, active surveillance is being conducted for potentially life-threatening, moderate-to-severe, and other serious adverse events and for vaccinia transmission to contacts of vaccinees (Table). Nonserious events are reported through passive surveillance and are expected to be underreported. This report summarizes smallpox vaccine adverse events reported among civilians vaccinated as of February 21, 2003, and received by CDC from the Vaccine Adverse Event Reporting System (VAERS) as of February 24. PMID- 12625501 TI - International laboratory animal science. PMID- 12625502 TI - Bordetella bronchiseptica infection of rats and mice. AB - Bordetella bronchiseptica has long been associated with respiratory tract infections in laboratory research, food-producing, companion, and wildlife animal species. Its range of distribution also may include humans and contaminated inanimate environmental sources. Natural diseases due to B. bronchiseptica infections in laboratory rats and mice were described before many of the major pathogens of these hosts were discovered. To our knowledge, there are no recent reports of natural disease due to B. bronchiseptica in these species; as a result, some have questioned its role as a natural pathogen in murine hosts. We reviewed occurrence of natural B. bronchiseptica infections and present information gained from recent experimental infection studies in murine hosts. We also discuss the potential impact of natural B. bronchiseptica infections on research and methods of control. PMID- 12625503 TI - Species characterization of plasma nitrite/nitrate (NOx) concentration. AB - Experiments were designed to detect and determine differences between nitrite/nitrate concentration ([NOx]) in plasma across 15 species selected from seven classes of vertebrates. Blood collected in syringes was placed immediately into ethylenediaminetetraacetic acid (EDTA)-containing tubes and was centrifuged. Plasma [NOx] was determined by measurement of chemiluminescence. Across classes of vertebrates, baseline plasma [NOx] ranged from 0.6 to 171.3 nmol/ml. Mean +/- SD plasma [NOx] was highest in a fresh-water, jawless fish (lamprey, 95.5 +/- 9.1 nmol/ml) and lowest in a saltwater cartilaginous fish (skates, 1.1 +/- 0.4 nmol/ml). Both amphibians tested had a wide range in plasma [NOx], which was explained partly by temporal changes during the year. Within the mammalian class, plasma [NOx] ranged from 3.8 to 43.2 nmol/ml. Results of this study indicate that NO is detectable in plasma of all classes of vertebrates and that baseline concentration varies among species. PMID- 12625504 TI - Influence of postoperative analgesics on the development of neuropathic pain in rats. AB - Rodent models of neuropathic pain require extensive tissue manipulation to induce the lesion of interest which results in inflammation and postoperative pain that is unrelated to nerve injury per se. We sought to determine whether acute postoperative pain management affects the development of hallmark signs of neuropathic pain. Analgesic regimens (q 24 h x 3 days) were buprenorphine (0.05 and 0.1 mg/kg of body weight, s.c.), flunixin meglumine (1.1 and 2.5 mg/kg, s.c.), and fentanyl citrate (0.01 and 0.1 mg/kg, i.p.). The spared nerve injury model of neuropathic pain was used, and mechanical and cold allodynia as well as body weight gain were measured for 28 days. Buprenorphine and fentanyl alleviated mechanical sensitivity and prevented weight loss associated with the surgery (0 to 3 days), but opioid-related adverse effects were observed. Flunixin reduced wound inflammation and improved weight gain, but had no effect on nociceptive thresholds. Cold allodynia was unaltered by any treatment. By postoperative day 7, control and treatment groups did not differ with respect to weight gain or nociceptive thresholds. Our findings suggest that postsurgical inflammation and pain behavior can be ameliorated without substantially altering the long-term development of neuropathic pain, provided that the selection of agent(s) and treatment regimen(s) is appropriate and the neuropathic pain of interest is evaluated seven days after surgery. PMID- 12625505 TI - Hematologic and serum biochemical values for zebrafish (Danio rerio). AB - The zebrafish (Danio rerio) has proven an excellent model for study of vertebrate development and genetics. Mutagenesis studies have produced many blood mutants with defects ranging from hematopoiesis to coagulation. The overwhelming majority of zebrafish studies have focused on development and mutational effects in embryos, whereas effects in mature zebrafish have gone largely unexplored. We believe that zebrafish will prove a valuable model for study of aging and age related diseases, and we have sought to characterize some of the basic features of mature zebrafish. Accordingly, blood was collected from adult zebrafish and was analyzed to determine reference hematologic and biochemical parameters. White blood cell differential counts indicated predominantly lymphocytes, with mean proportion of 82.95%. Total red blood cell counts averaged 3.02 x 10(6) cells/microl. Except for increases in alanine transaminase (ALT), amylase, and phosphorus values, serum biochemical analytes were within the range of reported values for mammals and other species of fish. Accurate analysis of the many zebrafish mutants generated requires determination of normal characteristics of zebrafish. We believe results such as these will help define normal adult zebrafish, which have a tremendous potential for use in the study of human disease and aging. PMID- 12625506 TI - Porcine model of diabetic dyslipidemia: insulin and feed algorithms for mimicking diabetes mellitus in humans. AB - A weakness of many animal models of diabetes mellitus is the failure to use insulin therapy, which typically results in severe body wasting. Data collected from such studies must be interpreted cautiously to separate the effects of hyperglycemia from those of starvation. We provide several algorithms that were used by us in two long-term (20-week) experiments in which hyperglycemia (300 to 400 mg/dl), dyslipidemia (cholesterol [280 to 405 mg/dl] and triglycerides [55 to 106 mg/dl] concentrations), and positive energy balance were maintained in swine. Yucatan miniature swine groups included control, alloxan-induced diabetes mellitus, diabetes mellitus plus diet-induced dyslipidemia, and exercise-trained diabetic dyslipidemic pigs. The algorithms were developed for the porcine model because of several similarities to humans, including: cardiac anatomy and physiology, propensity for sedentary behavior, and metabolism of dietary carbohydrates and lipids. Acute toxic effects of alloxan (hypoglycemia, hyperglycemia, nephrotoxicosis) were minimized by preventive fluid loading and by use of algorithms in which insulin, food, and fluid therapy were administered. Long-term insulin and food maintenance algorithms elicited normal body weight gain in all three diabetic groups (lean experiment) and threefold greater body weight gain in pigs of an obesity experiment. Exercise-trained pigs of both experiments manifested significantly increased work performance and did not experience medical complications. We conclude that these algorithms can be used in swine, or similar algorithms can be developed for other animal species to maintain hyperglycemia and/or dyslipidemia, while avoiding diabetes-induced wasting. Importantly, animal models of diabetes mellitus that maintain positive energy balance and poor glycemic control provide a marked improvement over other models by more closely mimicking the human presentation of diabetes mellitus. PMID- 12625507 TI - Hyperglycemia-induced insulin resistance in diabetic dyslipidemic Yucatan swine. AB - Hyperglycemia, dyslipidemia, and associated insulin resistance are hallmarks of diabetes mellitus. Purposes of the study reported here were to develop practical methods for assessment of in vivo insulin sensitivity and determine contributions of hyperglycemia and dyslipidemia to insulin resistance in the porcine model of alloxan-induced diabetes mellitus and dyslipidemia. Male Yucatan swine groups were treated for 20 weeks: control (C), high fat-fed (2% cholesterol) hyperlipidemic (H), alloxan-induced diabetic normolipidemic (D), diabetic high fat-fed (diabetic dyslipidemic, DD), and diabetic dyslipidemic treated with the lipid-lowering agent atorvastatin (DDA). Plasma cholesterol concentration increased sixfold in animals of groups H, DD, and DDA, whereas triglyceride concentration increased threefold in animals of group DD only. Diabetics had decreases in glucose tolerance and pancreatic immunostaining for insulin. Use of the gold standard hyperinsulinemic euglycemic clamp procedure indicated that maximal insulin-stimulated glucose uptake was similar to that in humans, but this method was not practical for use in pigs. Instead, a more convenient and valid insulin sensitivity test involving suppression of insulin secretion with somatostatin and a single insulin injection was used. Insulin sensitivity was greatly impaired by anesthesia with isoflurane, but was not affected by use of the anxiolytic agent diazepam. Insulin sensitivity decreased by 75% in diabetics (groups D, DD, DDA), compared with animals of groups C and H, and was inversely related to fasting blood glucose concentration (r = -0.72). Insulin treatment to restore blood glucose values of diabetics (> 250 mg/dl) to near control values (< 100 mg/dl) promptly restored insulin sensitivity to control values. We conclude that hyperglycemia is a major cause of insulin resistance in the porcine model of alloxan-induced diabetes mellitus and dyslipidemia. PMID- 12625508 TI - Detection of lymphocytic choriomeningitis virus by use of fluorogenic nuclease reverse transcriptase-polymerase chain reaction analysis. AB - Lymphocytic choriomeningitis virus (LCMV) induces persistent infections in laboratory mice; is a known contaminant of biological materials, such as transplantable tumor cell lines; and is of great concern in animal facilities due to its zoonotic potential. Fluorogenic nuclease reverse transcriptase-polymerase chain reaction (fnRT-PCR) assays combine RT-PCR with an internal fluorogenic hybridization probe, thereby potentially enhancing specificity and eliminating post-PCR processing. An fnRT-PCR assay specific for LCMV was, therefore, developed by targeting primer and probe sequences to a unique region of the LCMV nucleocapsid (NP) gene. The LCMV fnRT-PCR assay detected only LCMV and did not detect other RNA viruses that naturally infect rodents. The fnRT-PCR assay detected as little as one picogram of LCMV RNA, but was 100-fold less sensitive when directly compared with the mouse antibody production test. The fnRT-PCR assay was also able to detect viral RNA in numerous tissues and in feces and cage swipe specimens collected from experimentally inoculated BALB/c mice, but did not detect any viral RNA in similar samples collected from age- and strain-matched mock-infected mice. In conclusion, the LCMV fnRT-PCR assay offers a potentially high-throughput diagnostic assay to detect LCMV in mice and contaminated biological materials. PMID- 12625509 TI - Role of major histocompatibility complex class II in resistance of mice to naturally acquired infection with Syphacia obvelata. AB - Genetics plays a substantial role in host resistance in many host-parasite interactions. We examined the prevalence of naturally acquired infection with Syphacia obvelata in a number of mouse strains housed in a non-barrier facility. These mice, which included cross-bred and congenic, inbred strains on various genetic backgrounds, differ in the loci for the immune function genes--major histocompatibility complex class II (MHCII), toll-like receptor 4 (Tlr4), and solute carrier family 11, member 1 (Slc11a1)--which allowed comparisons of the impact of these genes on resistance to pinworm infection. Male and female mice of various ages were sampled over an 18-month period; infection was determined by use of the cellophane tape test. Results indicated that mice that were MHCII+/+ had a significantly lower prevalence of infection than did mice that were MHCII-/ . Differences were not seen between male and female mice. Although MHCII+/+ mice had an age-associated decrease in infection prevalence, such decrease was not seen in MHCII-/- mice. In contrast, infection prevalence in mice with the normal Tlr4 gene (Tlr4(LPS-n/LPS-n)) gene did not differ significantly compared with that in mice that were homozygous for either the point mutation (Tlr4(LPS-d/LPS d)) or deletion (Tlr4(LPS-del/LPS-del)) of that gene. Likewise, the presence (Sle11a1r/r) or absence (Slc11a1s/s) of functional alleles for Slc11a1 had no effect on the prevalence of infection with S. obvelata. In conclusion, presence of MHCII, but not Tlr4 or Slc11a1 significantly influences prevalence of naturally acquired infection with S. obvelata. These data justify further comprehensive analyses of the immune components that are involved in pinworm resistance. PMID- 12625510 TI - Switching of melanocyte pigmentation associated with pituitary pars intermedia tumors in Rb+/- and p27-/- female mice with yellow pelage. AB - As an incidental finding in a study of mammary tumorigenesis, two lines of genetically engineered mice were observed to develop pigmentation changes of the fur. Mice with targeted mutations of the Rb1 (Rb) and Cdkn1b (p27kip1) genes were crossed from C57BL/6 (black coat color; eumelanin) and 129Sv (wild-type agouti coat color) backgrounds, respectively, to one with a dominant yellow coat color (phaeomelanin) carrying a transgene for Agouti under a keratinocyte specific promoter. Both Rb+/- and p27-/- mice developed pituitary tumors of the pars intermedia that were associated with a switch to black (eumelanic) fur but were not observed in sibling Rb+/+ and p27+/+ mice. This phenomenon was observed first in the vibrissae and, subsequently one to two weeks later, as periorbital and dorsal patches, and was associated with pituitary lesions larger than four millimeters in the longest dimension. In Rb+/- mice, pigmentation change preceded a moribund state attributable to the tumors by two to four weeks, whereas in p27 /- mice, the pigmentation alteration was earlier, more gradual, and prolonged. The switch from phaeomelanin to eumelanin in the fur is most likely due to out competition of the agouti gene product by alpha-melanocyte-stimulating hormone from the pituitary tumors, an effect masked in black or agouti mice. PMID- 12625511 TI - Cryptosporidiosis associated with emaciation and proliferative gastritis in a laboratory-reared South African clawed frog (Xenopus laevis). AB - A 2-year-old emaciated female South African clawed frog (Xenopus laevis) was euthanized because of chronic weight loss. At necropsy, there was no evidence of bacterial, fungal or viral disease; however, the histopathologic findings indicated a proliferative gastritis and the presence of numerous cryptosporidial stages throughout the intestinal tract. Crytosporidial oocysts were present in the water taken from the aquarium housing the infected frog and were likely shed by the sick frog; however, the exact source of the oocysts could not be identified. Water samples from other frog aquaria in the facility did not contain cryptosporidial oocysts. Some Cryptosporidium species are important zoonotic pathogens and, to our knowledge, this is the first report of disease associated with Cryptosporidium infection in a laboratory Xenopus laevis. PMID- 12625512 TI - Administration of vaccinia virus to mice may cause contact or bedding sentinel mice to test positive for orthopoxvirus antibodies: case report and follow-up investigation. AB - Routine testing of bedding sentinels from a barrier room revealed one mouse seropositive to ectromelia virus (EV). Results of hemagglutination-inhibition testing and western blot analysis were confirmatory for orthopoxvirus antibodies. Additional seropositive animals were not identified. Interviews indicated that replication-competent vaccinia virus (VV), Western Reserve strain (VV-WR), recently had been given to mice. Although VV-WR was not expected to spread by contact or via fomites, the case evidence suggested transmission of vaccinia via soiled bedding. In a follow-up experiment, 15 index mice were inoculated with 10(7) plaque-forming units of VV by either subcutaneous or intrarectal instillation. A dedicated contact sentinel and a bedding sentinel were provided for each index mouse. All 15 index mice were positive for antibodies when tested 22 days after inoculation. One mouse, inoculated by the subcutaneous route, appeared ill and developed lesions on the proximal portion of the tail. The contact sentinel mouse housed with this index mouse was the only sentinel to seroconvert. We conclude that VV-WR can spread to contact sentinels and potentially to bedding sentinels. The ability of other VV strains to be transmitted horizontally and the susceptibility of different mouse strains to infection merit further investigation. The use of VV in animal facilities must be managed carefully since the available serologic tests do not distinguish between VV and EV, an exotic agent of major concern to laboratory animal facilities. PMID- 12625513 TI - Quality of life and coping with schizophrenia symptoms. AB - The question addressed in this paper is whether different coping styles mediate the relationship between psychopathology and related distress and the quality of life (QOL) among patients with schizophrenia. In a cross-sectional design, 161 schizophrenia inpatients were comprehensively evaluated with standardized measures of QOL, psychopathology, psychological distress and coping styles. Correlations and regression analyses were performed to examine the relationship among parameters and to estimate the mediating effect of coping styles on QOL in the framework of a distress/protection model. Life quality correlated positively with task- and avoidance-oriented coping styles and slightly negatively with emotion-oriented coping. Emotion-oriented coping mediated the relationship between the severity of activation, anxiety/depression symptoms, and QOL, while avoidance-oriented (distraction) coping was mediated between QOL and paranoid symptoms. Coping styles accounted for 25% of the variance in subjective QOL scores compared with 15% for psychological distress, and only 3% for clinical variables. The ability to cope with symptoms and associated distress substantially contributes to QOL appraisal in schizophrenia. Thus, different coping strategies may reduce the negative influence of specific symptoms and related distress on the subjective QOL of schizophrenia patients. PMID- 12625515 TI - Factors that contribute to quality of life outcomes prioritised by people with multiple sclerosis. AB - The aim was to investigate factors associated with depression and social function, two outcomes identified as important by people with multiple sclerosis (MS) and to identify underlying dimensions of psycho-social well-being that may be useful as outcome measures. People with MS in eight randomly selected health authorities/boards in England and Scotland completed a postal questionnaire relating to preferences and needs for their health and social care, along with the Beck Depression Inventory and the SF-36. Responses to 10 of the original items were subjected to factor analysis. These and other explanatory variables were entered into multivariable regression models for the two outcomes. The factor analysis resulted in three dimensions representing different aspects of psycho-social well-being; one of these (representing autonomy) was associated with improvements in both outcomes, as was the SF-36 emotional role limitation score. Three other SF-36 dimensions and lack of contact with a health professional in the last year were related just to social function. The regression models emphasise the value of enabling autonomy and self-reliance amongst people with MS, as well as more general measures of emotional health. The present work identifies specific questions that could be used to measure pivotal aspects of an individual's psycho-social well-being. While these findings warrant replication for people with MS, they may have relevance to those with other long term illnesses. PMID- 12625514 TI - Adjusting distributions of the Health Utilities Index Mark 3 utility scores of health-related quality of life. AB - The Health Utilities Index Mark 3 (HUI3) is a multi-dimensional, preference-based measure of health status and health-related quality of life (HRQoL). HUI3 scores correlate strongly with self-ratings of health status and functional disability and vary according to age, gender and occupation. In comparative studies relating to HRQoL, it is necessary to carry out adjusted comparison of the health status of the different groups. taking into account unbalanced distribution of confounding variables. This paper describes a stratification method to adjust the distributions of HUI3 scores. This method provides a graphical representation of adjusted distribution of HUI3, which can also be used to adjust other HRQoL scores. Cross-sectional data from the 1998/1999 National Population Health Survey (NPHS) in Canada were used to verify the proposed method. Male agriculture workers and male construction workers in Canada had quite different age distributions but similar HUI3 distributions. After adjusting the age distribution of the construction group to match the distribution of agriculture group, the mean HUI3 score of the former significantly decreased. PMID- 12625516 TI - Psychometric characteristics of the General Well-Being Schedule (GWB) with African-American women. AB - The General Well-Being Schedule (GWB) is a brief, reliable, and valid instrument used in population studies to assess psychological well-being, although its validity with African-Americans has yet to be established. This study evaluated the reliability, validity, and factor structure of the GWB in a sample of 599 overweight African-American women who participated in multicenter weight loss trial. The results of the factor analysis indicate that the GWB is primarily unidimensional and that the existence of the six hypothesized subscales was not supported. The GWB demonstrated evidence of concurrent and construct validity when examined in association with measures of self-concept, depression, and several health behaviors. The results of this study suggest that the GWB is a reliable and valid measure of psychological well-being in African-American women. PMID- 12625517 TI - The mediating role of optimism on post-radiation quality of life in nasopharyngeal carcinoma. AB - Nasopharyngeal carcinoma (NPC) is prevalent in southern China. In non-welfare countries, eating difficulties after radiotherapy may hinder NPC patients' short term quality of life (QoL) especially regarding work and finance. This study explored the positive mediating role of optimism on the relationship between eating ability and QoL of Chinese NPC patients who received radiotherapy in Hong Kong. A sample of 211 newly referred patients were recruited and followed-up face to-face or by telephone at four (post-radiation 1) (FU 1) and eight (post radiation 2) (FU 2) months from baseline. QoL was measured by the FACT-G (Chinese). Optimism and eating ability were measured by two 11-point self-rated items. Mediating effects were tested with a series of multiple regression models. After adjustment for pre-radiation QoL, socioeconomic status and cancer stage, optimism significantly mediated the relationship between eating ability at FU 1 and overall QoL at FU 2, in particular physical and social/family well-being. This study is the first to underpin the mediating role of optimism on QoL of NPC patients. Faced with aversive side-effects of radiotherapy, facilitating positive thinking in Chinese NPC patients may accelerate their short-term post-radiation adjustment. PMID- 12625518 TI - Quality of life and menopausal transition for middle-aged women on Kinmen island. AB - Although it seems reasonable to suggest that most women experience significant changes in quality of life (QOL) during the menopausal period, few researchers have quantified these changes. A total of 1497 women, aged 40-54 years and living on the island of Kinmen, were recruited for this survey. However, 137 were eliminated leaving 1360 for analysis. Women who used hormonal therapy or who had undergone surgically induced menopause were excluded. The subjects with incomplete data or who exhibited mental retardation or severe psychiatric disease were also eliminated. Univariate analysis demonstrated that, in general, QOL scores were poorer for perimenopausal and postmenopausal status. Comparing pre- and postmenopausal women, significant statistical differences were demonstrated for role limitations due to physical and emotional problems, even after adjusting for age, education level, body mass index, menarche, and chronic illness. A strong association was demonstrated between menopausal symptoms and premenstrual syndrome (PMS). Women with menopausal symptoms and PMS had significantly lower scores on all SF-36 dimensions. The results of this study suggest that poorer health status is experienced by peri- and postmenopausal women compared to premenopausal women. PMID- 12625519 TI - Development of the Cystic Fibrosis Questionnaire (CFQ) for assessing quality of life in pediatric and adult patients. AB - To assess the impact of cystic fibrosis (CF) and treatment on quality of life (QOL) from childhood throughout adult age, two versions of the Cystic Fibrosis Questionnaire (CFQ), were developed and validated in France: the CFQ 14+ for teenagers and adults, the CFQ Child P, a parent-proxy evaluation for children aged 8-13. They include three modules for assessing QOL, symptoms and health perception. Nine QOL dimensions were identified: physical functioning, energy/well-being, emotions, social limitations, role, embarrassment, body image, eating disturbances and treatment burden. Items were derived from 33 interviews with patients and parents. Item reduction and assessment of internal consistency, convergent and discriminant validity were based on a large cross-sectional survey among 393 patients and parents. A second study was conducted among 124 patients and 85 parents to test reproducibility and responsiveness, confirm the subscale structure and assess scalar properties using Rasch analysis. All psychometric properties were successfully demonstrated and both the CFQ 14+ and the CFQ Child P French questionnaires are now well validated. German and Spanish validated adaptations are available, an English validation is in progress. CONCLUSION: The CFQ 14+ and CFQ Child P are well validated, multilingual measures which allow QoL assessment in children, teenagers and adults with CF. PMID- 12625520 TI - The revised German Cystic Fibrosis Questionnaire: validation of a disease specific health-related quality of life instrument. AB - BACKGROUND: The assessment of health related quality of life (HRQOL) has increasingly been recognized as an important adjunct to medical outcome parameters in the monitoring of cystic fibrosis (CF) patients and in clinical studies. The Cystic Fibrosis Questionnaire (CFQ) is a disease-specific HRQOL instrument for CF patients developed in France. Translations are currently being validated in four different countries. The aim of the present study was to validate the German adaptation of the CFQ-14+. METHOD: On the basis of results from a first dataset (n = 197 CF adolescent and adult patients), revisions were made and retested in a second sample of 103 patients. RESULTS: The final revised version showed good construct validity. The same nine HRQOL domains as in the French original CFQ-14+ emerged from the analyses as structurally robust scales. The internal consistencies of the HRQOL scales ranged from 0.71 to 0.94. The instrument's clinical validity was supported by severely ill patients reporting lower HRQOL than less severely ill patients on most scales. CONCLUSION: We conclude that the evidence supports the validity and reliability of the instrument. An important future area of application is the use in comparative multi-center international studies. PMID- 12625521 TI - Validity and reliability of the EQ-5D self-report questionnaire in English speaking Asian patients with rheumatic diseases in Singapore. AB - Validity and reliability of a Singaporean English EQ-5D self-report questionnaire (EQ-5D) were evaluated among consecutive outpatients with rheumatic diseases attending a tertiary referral hospital in Singapore (a multi-ethnic, urban Asian country). Subjects were interviewed twice within a 2-week period using a standardized questionnaire containing the EQ-5D, Short Form 36 Health Survey (SF 36) and assessing demographic and psychosocial characteristics. To assess validity of the EQ-5D, 13 hypotheses relating responses to EQ-5D dimension/Visual Analogue Scale (EQ-VAS) to SF-36 scores or other variables were examined using the Mann-Whitney test, Kruskal-Wallis test, or Spearman's correlation coefficient. Test-retest reliability was assessed using Cohen's kappa. Sixty-six subjects were studied (osteoarthritis: 9, rheumatoid arthritis: 26, systemic lupus erythematosus: 23, spondyloarthropathy: 8; female: 72.7%; mean age: 44.3 years). Ten of 13 a-priori hypotheses relating EQ-5D responses to external variables were fulfilled, supporting the validity of the EQ-SD. Cohen's kappa for test-retest reliability (n = 52) ranged from 0.29 to 0.61. The Singaporean English EQ-5D appears to be valid in measuring quality of life in Singaporeans with rheumatic diseases; however, its reliability requires further investigation. These data provide a basis for further studies assessing the validity of the EQ 5D in Singapore. PMID- 12625522 TI - Quality of life in patients of nasopharyngeal carcinoma: validation of the Taiwan Chinese version of the EORTC QLQ-C30 and the EORTC QLQ-H&N35. AB - The authors followed the guidelines of translation and pilot testing of the EORTC QLQ-C30 and EORTC-QLQ-H&N35 questionnaires. The questionnaires were given to 50 nasopharyngeal carcinoma patients under active treatment and 50 under follow-up at our institution from November 2000 to June 2001. A retest was conducted 2 weeks after the first interview/form completion for the follow-up group. The intraclass correlation coefficients of the two questionnaires were moderate to high in the follow-up group. Cronbach's alpha coefficients of all scales of the two questionnaires were > or = 0.70 except that of cognitive functioning. Correlation of scales measuring similar dimensions of the QLQ-C30 and the SF-36 were moderate to high, while that of the QLQ-H&N35 and the QLQ-C30 and the SF-36 were moderate to low. Patients in the active treatment group had more serious acute problems due to disease and chemotherapy. Patients in the follow-up group had more serious chronic problems due to radiation therapy. We concluded that the Taiwan Chinese version of the EORTC QLQ-C30 and the EORTC QLQ-H&N35 had moderate to high test-retest reliability, high internal consistency in most scales, and could show the expected differences between patients in active treatment and follow-up group. PMID- 12625523 TI - Free oxygen radicals are generated at the time of aspiration of oocytes from ovaries that have been stored for a long time. AB - In ischaemic tissues, reperfusion induces acute injury and functional changes. In this work, ovaries were stored for various times, and superoxide dismutase (SOD) and dimethylthiourea (DTMU) were used at the time of oocyte aspiration. We then attempted to determine whether free oxygen radicals are generated at oocyte aspiration and whether they impair the developmental competence of oocytes. Over 2 mM of DMTU and 1000 U/ml of SOD significantly improved the rate of blastulation 8 days after insemination. For ovaries that were preserved for 3 and 7 h, using antioxidants also significantly improved the rate of blastulation 8 days after insemination. However, no effect was observed on oocytes from ovaries preserved for 1 h. We examined how the antioxidants affected the presence of germinal vesicles, chromatin configuration, and polar body extrusion 6 or 21 h after culture. Chromatin configuration was classified into three groups according to the amount of chromatin condensation (group 1, strong condensation; group 2, moderate; group 3, slight). Storing ovaries for a long time decreased the frequency of occurrence of group 2, but increased groups 1 and 3. However, using antioxidants at oocyte aspiration decreased the frequency of group 3 and increased group 1. Moreover, there was no difference in the rate of germinal vesicle breakdown and polar body extrusion. Our results show that preserving ovaries for a long time induces the generation of free oxygen radicals and that these chemicals impair oocyte viability. Using antioxidants at oocyte aspiration was beneficial for embryo production. PMID- 12625524 TI - MAPK/ERK kinase (MEK) signalling is required for resumption of meiosis in cultured cumulus-enclosed pig oocytes. AB - Resumption of meiosis of mammalian oocytes is facilitated by the maturation promoting factor (MPF) and accompanied by activation of mitogen activated protein kinases (MAPK) which are phosphorylated by the MAPK kinase (MEK). In this study we examined the effects of PD 98059, which inhibits the activity of MEK, on in vitro maturation of pig oocytes. Cumulus-oocyte complexes (COCs) were cultured in the presence or absence of the drug (50 microM) for various time periods. To elucidate the influence of cumulus cells, COCs were first cultured in inhibitor free medium, subsequently denuded, and incubated further in PD 98059 supplemented medium. Reversibility of drug action as tested following PD 98059 treatment of COCs by transferring them to drug-free medium. Culture of COCs in medium supplemented with PD 98059 prevents resumption of nuclear maturation in the majority of COCs. This inhibition was reversible and accompanied by a non activation of both MAP and MPF. Addition of the MEK inhibitor to extracts of in vitro matured oocytes revealed that the kinase activities were not directly influenced by the inhibitor, suggesting a link between MAP and MPF kinases. Preincubation of COCs in inhibitor-free medium for 6 h followed by further culture of COCs or denuded oocytes in the presence of PD 98059 for various periods resulted in elevated MAP and MPF kinase activities, indicating an early and transient MEK signalling in the oocyte itself. These results support the idea that MAP and MPF are involved in the induction of germinal vesicle breakdown in porcine oocytes. PMID- 12625525 TI - Sex determination of buffalo embryos (Bubalus bubalis) by polymerase chain reaction. AB - The aim of this study was to identify a simple, rapid method for sex determination of in vitro produced buffalo embryos, amplifying Y-chromosome specific repeat sequences by polymerase chain reaction (PCR). Buffalo oocytes collected from slaughtered animals were matured, fertilised and cultured in vitro for 7 days. On day 7 embryos were evaluated and divided in to six groups according to developmental stage (2, 4, 8, 16 cells, morulae and blastocyst). Each embryo was stored singly in phosphate-buffered saline at -20 degrees C until PCR. Two different methods of extraction of DNA were compared: a standard procedure (ST), using a normal extraction by phenol-chloroform, isoamyl alcohol and final precipitation in absolute ethanol and a direct procedure (DT), using a commercial kit (Qiaquik-Qiagen mini blood). A pair of bovine satellite primers and two pairs of different bovine Y-chromosome-specific primers (BRY4.a and BRY.1) were used in the PCR assay on embryos and on whole blood samples collected from male and female adult buffaloes, used as control. The trial was carried out on 359 embryos (193 for ST and 166 for DT). When DNA samples from blood were amplified, the sex determined by PCR always corresponded to the anatomical sex. Embryo sexing was not possible in two embryos in ST and one embryo in DT. Both extraction protocols recovered sufficient quantities of target DNA at all developmental stages, but the time required for the ST (24 h) limits its use in embryo sexing and supports the use of commercial extraction kits (5 h). PMID- 12625526 TI - Review of the longevity of the second polar body in the mouse. PMID- 12625527 TI - DNase I and II present in avian oocytes: a possible involvement in sperm degradation at polyspermic fertilisation. AB - During polyspermic fertilisation in birds numerous spermatozoa enter the eggs, in contrast to the situation in mammals where fertilisation is monospermic. However, in birds only one of the spermatozoa which have entered an egg participates in zygote nucleus formation, while the supernumerary spermatozoa degenerate at early embryogenesis. Our previous work has demonstrated the presence in preovulatory quail oocytes of DNase I and II activities able to digest naked lambdaDNA/HindIII substrate in vitro. In the present studies, the activities of both DNases in quail oocytes at different stages of oogenesis and in ovulated mouse oocytes were assayed in vitro using the same substrate. Degradation of quail spermatozoa by quail oocyte extracts was also checked. Digestion of the DNA substrate was evaluated by electrophoresis on agarose gels. The activities of DNase I and II in quail oocytes increased during oogenesis and were the highest in mature oocytes. The activities were present not only in germinal discs but also in a thin layer of cytoplasm adhering to the perivitelline layer surrounding the yolk. At all stages of oogenesis the activity of DNase II was much higher than that of DNase I. DNA contained in spermatozoa was also degraded by the quail oocyte extracts under conditions optimal for both DNases. In contrast to what is observed in quail oocytes, no DNase activities were detected in ovulated mouse eggs; this is logical as they would be useless or even harmful in monospermic fertilisation. The possible role of DNase activities in avian oocytes, in degradation of accessory spermatozoa during polyspermic fertilisation, is discussed. PMID- 12625528 TI - Identification of differentially expressed mRNAs in bovine preimplantation embryos. AB - We have characterised the changes in preimplantation embryos that occur in the mRNA population during the transition from maternal to zygotic control of embryogenesis. We connected the mRNA differential display method and RT-PCR based method that allows amplification of the whole population of messengers. In the early stages of development we have further characterised the level of individual mRNAs with the help of semiquantitative RT-PCR used with specific primers. This report concerns four of 12 cDNA fragments that appeared to be differentially expressed between the 4- and late 8-cell stages. A transcript corresponding to fragment no. 1/12 appears to be analogous to the maternal mRNA since it is abundant in 1-, 2-, 4- and 8-cell embryos and rapidly decreases in the later stages. A similar pattern of expression was revealed in the transcript corresponding to fragment no. 8/9. A transcript corresponding to fragment no. 20/8 is newly synthesised from the embryonic genome at the late 8-cell stage and its amount rapidly increases during the following stages. This messenger shows a 91.7% identity with mRNA for human S3A ribosomal protein and 92.2% identity with mRNA for Felis domesticus S3A ribosomal protein. A transcript corresponding to fragment no. 8/19 is stage-specific, being newly synthesised from an embryonic genome at the late 8-cell stage and decreasing in the later stages. This messenger shows 86.6% identity with a mouse mRNA for proline-rich protein and 91.6% identity with human mRNA for KIAA-0058 gene. A complex of these molecular markers represents a suitable tool for answering questions concerning the molecular control of major gene activation during bovine embryogenesis. PMID- 12625529 TI - Development of mouse embryos derived from oocytes reconstructed by metaphase I spindle transfer. AB - Abnormal oocyte spindle is frequently associated with the infertility of aged women. Directly manipulating the metaphase I (MI) spindle may be a feasible method to overcome this kind of problem. Here, we report that the MI meiotic spindle can be removed from MI mouse oocytes and will autonomously divide into two daughter cells with the same size, morphology and an equal number of chromosomes after culture for 5 h in maturation medium. The division rate of the MI spindle reached 56% after 10-15 h of culture. After transferring the MI meiotic spindle into synchronous ooplasm by electrofusion, about 61% of the reconstructed oocytes continued to complete the first meiosis and extruded a normal first polar body. The matured reconstructed oocytes can also be fertilised. Approximately 50% of the 2-cell embryos developed to the morula stage after in vitro culture. PMID- 12625530 TI - The effect of PD98059 on MAPK regulation in cumulus-enclosed and cumulus-free mouse oocytes. AB - The effect of the p42/44 mitogen-activated kinase (MAPK) inhibitor, PD98059, on MAPK activation and meiosis resumption in mouse oocytes was studied. When germinal vesicle (GV)-stage denuded oocytes (DOs) were cultured continuously in 50 microM PD98059, germinal vesicle breakdown (GVBD) was postponed for 2-3 h. MAPK phosphorylation and activation was delayed as well. However, PD98059 did not impair histone H1 kinase activation. After 14 h of culture there was no significant difference in the rate of DOs reaching metaphase II (MII) arrest in either control or experimental conditions. The effect of PD98059 on MAPK inhibition was further tested in epidermal growth factor (EGF)-treated oocytecumulus complexes (OCCs). Exposure of GV-stage OCCs for 5 min to EGF (10 ng/ml) induced a considerable increase in MAPK phosphorylation. After OCCs were further cultured in 50 microM PD98059 a rapid dephosphorylation of MAPK was induced. Already after 1 min of treatment the non-phosphorylated form of MAPK dominated, indicating the high effectivity of PD98059. This result indicates that short EGF/PD98059 treatment of OCCs induced MAPK phosphorylation/dephosphorylation in cumulus cells only. As only a transient delay in MAPK phosphorylation and activation was observed in PD98059-treated DOs we conclude that there is also another PD98059-nonsensitive pathway(s) leading to MAPK activation in mouse oocytes. The data obtained suggest that meiosis resumption in mouse oocytes is somehow influenced by the MEK/MAPK activation pathway. PMID- 12625531 TI - Different activation treatments for successful development of bovine oocytes following intracytoplasmic sperm injection. AB - In this study, the developmental capacity and cytogenetic composition of different oocyte activation protocols was evaluated following intracytoplasmic sperm injection (ICSI) of in vitro matured bovine oocytes. Motile spermatozoa selected by Percoll density gradient were treated with 5 mM dithiothreitol (DTT) and analysed for ultrastructural changes of the head using transmission electron microscopy (TEM). The alterations in sperm morphology after DTT treatment for different times (15, 30 and 60 min) were 10%, 45-55% and 70-85%, respectively. Further, a partial decondensation of sperm heads was observed after DTT treatment for 30 min. Oocytes were injected with sperm treated with DTT for 30 min. In group 1, sperm injection was performed without any activation stimulus to the oocytes. In group 2, sham injection without sperm was performed without activating the oocytes. Oocytes injected with sperm exposed to 5 microM ionomycin for 5 min (group 3), 5 microM ionomycin + 1.9 mM dimethylaminopurine (DMAP) for 3 h (group 4) and 5 microM ionomycin + 3 h culture in M199 + 1.9 mM DMAP (group 5) were also evaluated for cleavage, development and chromosomal abnormality. Cleavage and development rates in groups 1, 2 and 3 were significantly (p < 0.05) lower than those in groups 4 and 5. The incidence of chromosomal abnormality in the embryos treated directly with DMAP after ionomycin (group 4) was higher than in group 5. We conclude that immediate DMAP treatment after ionomycin exposure of oocytes results in arrest of release of the second polar body, and thus leads to changes in chromosomal pattern. Therefore, the time interval between ionomycin and DMAP plays a crucial role in bovine ICSI. PMID- 12625532 TI - In vitro development of equine nuclear transfer embryos: effects of oocyte maturation media and amino acid composition during embryo culture. AB - This study was conducted to evaluate the effects of insulin-like growth factor I (IGF-I) and other media factors during oocyte maturation, and the presence of different compositions of amino acids in embryo culture medium, on the development of equine embryos. Oocytes recovered from slaughterhouse-derived ovaries were matured in vitro for 24 h and those with a polar body were subjected to intracytoplasmic sperm injection (ICSI) or nuclear transfer with adult fibroblasts (NT). For ICSI embryos, there were no significant differences in rates of morphological cleavage, cleavage with normal nuclei or average nucleus number at 96 h post-ICSI between the absence and presence of IGF-I in maturation medium, or between embryos cultured in G1.2 or a modified CZB medium (CZB-C). Embryos produced by interspecies NT (equine donor cells into bovine cytoplasts) also showed no difference in cleavage rate or average nucleus number whether cultured in G1.2 or in CZB-C. The rates of cleavage, cleavage with normal nuclei and average nucleus number of equine NT embryos were not significantly different among oocytes matured in M199 with FSH in the presence or absence of IGF-I, or in EMMI medium, which contains IGF-I, epidermal growth factor, steroid hormones, FSH and LH. There were no differences in development of equine NT embryos cultured in any of three amino acid treatments (with or without non-essential amino acids, or containing taurine, hypotaurine and cysteine only). The cleavage rate and average nucleus number of parthenogenetically activated oocytes (treated similarly to NT oocytes but not enucleated or subjected to donor cell injection) were significantly (p < 0.05) higher than those for NT embryos. These results indicate that the presence of IGF-I or of EMMI medium during in vitro maturation of equine oocytes does not have a beneficial effect on their developmental competence as assessed at 96 h. Presence or absence of non-essential amino acids in embryo culture medium does not affect development of NT embryos within the first 96 h of culture. Factors associated with enucleation or nuclear transfer decrease the developmental competence of equine NT embryos. CZB-C medium may be used for culture of equine embryos with results similar to those obtained with G1.2 medium, thus providing a base medium that may be modified for further study of culture requirements of equine embryos. PMID- 12625533 TI - Frequency of sex chromosomal mosaicism in bovine embryos and its effects on sexing using a single blastomere by PCR. AB - Assessment of nuclear status is important when a biopsied single blastomere is used for embryo sexing. In this study we investigated the nuclear status of blastomeres derived from 8- to 16-cell stage in vitro fertilised bovine embryos to determine the representativeness of a single blastomere for embryo sexing. In 24 embryos analysed, the agreement in sex determination between a biopsied single blastomere and a matched blastocyst by polymerase chain reaction (PCR) was 83.3%. To clarify the discrepancies, karyotypes of blastomeres in 8- to 16-cell stage bovine embryos were analysed. We applied vinblastine sulfate at various concentrations and for different exposure times for metaphase plate induction in 8- to 16-cell stage bovine embryos. The 1.0 mg/ml vinblastine sulfate treatment for 15 h was selected as the most effective condition for induction of a metaphase plate (> 45%). Among 22 embryos under these conditions, only 8 of 10 that had a normal diploid chromosome complement showed a sex chromosomal composition of XX or XY (36.4%) and 2 diploid embryos showed mosaicism of the opposite sex of XX and XY in blastomeres of the embryo (9.1%). One haploid embryo contained only one X-chromosome (4.5%). Four of another 11 embryos with a mixoploid chromosomal complement contained a haploid blastomere with a wrong sex chromosome (18.2%). In conclusion, assessment of nuclear status of 8- to 16-cell stage bovine embryos revealed that morphologically normal embryos had a considerable proportion of mixoploid blastomeres and sex chromosomal mosaicism; these could be the cause of discrepancies in the sex between biopsied single blastomeres and matched blastocysts by PCR. PMID- 12625534 TI - Antibiotic resistance and medicines licensing. PMID- 12625535 TI - Communicating quality--a key message for veterinary practice. PMID- 12625536 TI - Evaluation of once daily treatment with cyclosporine for anal furunculosis in dogs. AB - Twenty-four dogs with anal furunculosis were treated with cyclosporine once daily for 13 weeks at dosages of 1.5, 3.0, 5.0 or 7.5 mg/kg, and re-examined after six and 12 months. After 13 weeks the disease in six of the dogs was in remission, 11 were controlled or improved and seven had failed to respond. The response of the dogs given the highest dose was significantly better than the response of the other groups taken together (P < 0.014), and better than the responses of the groups given 1.5 mg/kg and 5 mg/kg (P < 0.05). The dogs improved clinically during the treatment, most rapidly during the first five weeks. Of the six dogs that were in remission after 13 weeks, three relapsed after one, two and six months. The 11 dogs that were improved or controlled after 13 weeks were either left untreated or were continued on cyclosporine medication for one to three months at a dosage of 1.5 to 7.5 mg/kg; the disease went into remission in four cases and remained controlled in the other seven, but four of the 11 cases relapsed during the 12 months following the treatment. The side effects observed included increased coat turnover and transient vomiting. PMID- 12625537 TI - Characterisation of Mycoplasma gallisepticum strains involved in respiratory disease in pheasants and peafowl. AB - Two cases of Mycoplasma gallisepticum infection in different avian species in backyard gamebird operations in Slovenia were investigated. In the first case, M gallisepticum was associated with severe respiratory disease with almost 20 per cent mortality in pheasants, whereas the infection was less pathogenic for chickens and turkeys reared at the same site. The M gallisepticum isolates from pheasants had a unique pMGA gene sequence containing a repeat of 12 nucleotides, and they contained only small amounts of the cytadhesins MGC1 and MGC3 and no PvpA protein. However, they expressed some typical M gallisepticum proteins and several proteins which were immunogenic for pheasants, chickens and turkeys. A strain of M gallisepticum isolated from the sinus of a pheasant was highly pathogenic for chicken embryos. In the second case, the M gallisepticum strain that was associated with respiratory disease and mortality in peafowl also affected chickens. M gallisepticum strain ULB 992 was isolated from the infraorbital sinus of a dead peafowl. The ULB 992 strain synthesised a small amount of MGC3, a truncated form of MGC1 and lacked PvpA. However, it expressed several proteins which were immunogenic for the birds infected with M gallisepticum at both gamebird operations. PMID- 12625538 TI - Efficacy of selamectin against biting lice on dogs and cats. AB - The efficacy of selamectin was evaluated against naturally acquired Trichodectes canis infestations on dogs and against Felicola subrostratus infestations on cats. Twenty dogs and 18 cats were randomly allocated to treatment with either a placebo or selamectin (6 mg/kg), administered topically once only on day 0. The treatment had no adverse effects in either the dogs or the cats. Efficacy was assessed by counting the live lice (adults and nymphs) on each animal by using a coat-parting technique on days -3, 7, 14, 21, 28, 35 and 42 for the dogs, and on days -1, 7, 21, 35 and 42 for the cats. On day 43, the number of live lice on each dog was also assessed by using a whole-body combing technique. Selamectin was 100 per cent effective in killing biting lice on the dogs and cats throughout the period of assessment; the louse counts on the treated dogs and cats were significantly lower than the pretreatment counts (P = 0.0001) and were also significantly lower than on the placebo-treated dogs (P < 0.05) and cats (P = 0.0001). There was a marked reduction in the prevalence of clinical signs associated with ectoparasite infestation in the treated dogs and no clinical signs were observed in any of the treated cats. PMID- 12625540 TI - Relationship between fertility and the walking activity of cows at oestrus. PMID- 12625539 TI - Canine distemper virus in river otters in British Columbia as an emergent risk for coastal pinnipeds. PMID- 12625542 TI - Student membership of the BVA. PMID- 12625541 TI - Outbreak of malignant catarrhal fever in cattle in Turkey. PMID- 12625543 TI - Export of dogs and cats to New Zealand. PMID- 12625544 TI - Fees for retired RCVS members. PMID- 12625546 TI - Veterinary nursing surveys. PMID- 12625545 TI - Revision of the Veterinary Surgeons Act. PMID- 12625547 TI - Control of status epilepticus. PMID- 12625548 TI - Expert witnesses. PMID- 12625549 TI - Solving chemical problems of environmental importance using cavity ring-down spectroscopy. AB - Cavity ring-down (CRD) is a sensitive variant of traditional absorption spectroscopy that has found increasing use in a number of chemical measurement applications. This review focuses on applications of cavity ring-down spectroscopy that will be of interest to environmental chemists and analytical chemists working on environmental problems. The applications are classified into direct monitoring approaches, indirect analysis methods and ancillary studies and a differentiation is made between field-tested instruments and proof of principle studies. PMID- 12625550 TI - Imaging TOF-SIMS analysis of oligonucleotide microarrays. AB - Single strand thiolated oligonucleotide (25-mer) was printed onto chemically modified glass and silicon surfaces. Confirmation of the level of attachment attained in each case was effected through detection by conventional confocal fluorescence microscopy. Both positive-ion and negative ion imaging time-of flight mass spectra were recorded for the visualization of micro-patterned oligonucleotide arrays. This represents the first report of such detection by this form of mass spectrometry on glass. Ultimately, we are interested in the possibility that imaging time-of-flight secondary ion mass spectrometry can discern the orientation and conformation of DNA strands present on the surface of a substrate. PMID- 12625551 TI - Detection and characterization of 20-oxosteroids in rat brains using LC-electron capture APCI-MS after derivatization with 2-nitro-4 trifluoromethylphenylhydrazine. AB - The detection and characterization of 20-oxosteroids in rat brains by liquid chromatography (LC)-electron capture atmospheric pressure chemical ionization (ECAPCI)-mass spectrometry (MS) is described. In the present method, oxosteroids were derivatized with a highly electron-affinitive reagent, 2-nitro-4 trifluoromethylphenylhydrazine, to convert them to the corresponding hydrazones. The derivatized steroids showed over 20-fold higher sensitivity in ECAPCI-MS than intact steroids measured in positive-APCI-MS. A brain sample was homogenized, purified with two disposable cartridges, derivatized and subjected to LC-ECAPCI MS. Pregnenolone, progesterone, 5alpha-dihydroprogesterone, allopregnanolone and epiallopregnanolone were clearly detected with a 50 mg sample of the brain tissue. PMID- 12625552 TI - Ligand exchange chiral separations by cyclodextrin derivatives in capillary electrophoresis. AB - A beta-cyclodextrin substituted by an imidazole-bound histamine (CDmh) was successfully used to separate underivatised tryptophan racemate in capillary electrophoresis in the presence of copper(II) ion by the ligand exchange mechanism. To the best of our knowledge, this is the first example of a cyclodextrin derivative behaving as the first coordination sphere ligand in the complex added to the background electrolyte (BGE). PMID- 12625553 TI - Assessment of the cross-reactivity and binding sites characterisation of a propazine-imprinted polymer using the Langmuir-Freundlich isotherm. AB - In this paper, the Langmuir-Freundlich isotherm is used to model the interaction of several triazines (desethylatrazine, desisopropylatrazine, simazine, atrazine, propazine and prometryn) with a propazine-imprinted polymer and to explain the observed cross-reactivity. Different rebinding experiments (each herbicide alone or all together in a mixture) were carried out and the experimental binding isotherms were fitted to the Langmuir-Freundlich isotherm. The fitting coefficients obtained (total number of binding sites, mean binding affinity and heterogeneity index) allowed the description of the kind of binding sites present in the imprinted polymer under study. It was concluded that the recognition mechanism was mainly governed by the molecular size although slight differences in the molecular structure may also play an important role. The obtained results suggest that the use of this new methodology can open new pathways for understanding how molecular recognition in imprinted polymers takes place. PMID- 12625554 TI - Investigation of the competitive rate of derivatization of several secondary amines with phenylisocyanate (PHI), hexamethylene-1,6-diisocyanate (HDI), 4,4' methylenebis(phenyl isocyanate) (MDI) and toluene diisocyanate (TDI) in liquid medium. AB - The stabilization of the isocyanate (NCO) groups during workplace sampling is necessary for their subsequent laboratory analysis. Most derivatization reagents are secondary amines. By carrying out a test in which two secondary amines are added to an isocyanate, the relative rates of these reactions can be evaluated. This evaluation is known for a monoisocyanate, phenylisocyanate (PHI), but is being developed for diisocyanates. This study deals with the relative reactivity (RR) of four diisocyanates: hexamethylene 1,6-diisocyanate (HDI), 4,4' methylenebis(phenyl isocyanate) (MDI), and the ortho and para isomers of toluene diisocyanate (TDI) in addition to PHI, with four secondary amines: 1-(2 methoxyphenyl)piperazine (MOPIP), 9-(N-methylaminomethyl)anthracene (MAMA), 1-(9 anthracenylmethyl)piperazine (MAP), and dibutylamine (DBA). These competitive derivatization reactions are studied in three reaction solvents, namely acetonitrile, toluene, and acetonitrile doped with water (1% v/v). The results show that the order of reactivity, which doesn't change with the isocyanate as well as with the solvent used, is the following: DBA > MAP > MOPIP > MAMA. The relative difference in reactivity is a function of both the isocyanate and the solvent used. Hindered aromatic diisocyanates (TDI and MDI) show a greater difference in reactivity with the derivatization agents. These differences in reactivity are also modified by the solvent used. For example, larger differences are observed in acetonitrile than in toluene, but the introduction of water to acetonitrile, which does not affect the reaction yield, makes these differences smaller. PMID- 12625555 TI - Stability of polycyclic aromatic hydrocarbons in pressurised hot water. AB - The stability of seven polycyclic aromatic hydrocarbons (PAHs) were studied in aqueous solutions at 100-350 degrees C. The aqueous solution of one PAH compound at a time was sealed in a stainless steel bomb-type reaction vessel in an argon atmosphere and heated for 10 to 240 min in an oven. The contents of the vessel were analysed by GC-MS. At 300 degrees C, degradation was observed even with the shortest heating time, 10 min. The degradation products were typically different oxidation products such as ketones and quinones. In addition to the heating time and temperature, the concentration of the analyte and the inner surface of the vessel affected the results. PMID- 12625556 TI - Studies on nitroaromatic compound degradation in modified Fenton reactions by electrospray ionization tandem mass spectrometry (ESI-MS-MS). AB - Degradation products resulting from modified Fenton reactions with the nitroaromatic compounds trinitrotoluene (TNT) and trinitrobenzene (TNB) were identified by electrospray ionization tandem mass spectrometry (ESI-MS-MS). Several hydroperoxide adducts were tentatively identified as initial, one electron reduction products of TNT and tandem mass spectrometry confirmed their structure. A transformation pathway of TNT, resulting from reactions with oxygen radical species generated by the modified Fenton reaction, was proposed. PMID- 12625557 TI - DNA hybridization in reverse micelles and its application to mutation detection. AB - DNA hybridization was investigated in AOT (sodium di-2-ethylhexyl sulfosuccinate)/isooctane reverse micelles. The single-stranded DNA molecules were encapsulated in the nanoscale water pools formed in the reverse micelles, reducing the hybridization rate. The DNA hybridization can be monitored by simply measuring the UV absorbance of the reverse micellar solution at 260 nm. We found that the DNA hybridization occurred only at the restricted water content (Wo = [H2O]/[AOT] = 20) and below room temperature. We applied this DNA hybridization behavior in reverse micelles to mutation detection in a model gene p53 and successfully detected the single nucleotide mutations in 20-mer. 30-mer and 50 mer nucleotides without a DNA labeling. PMID- 12625558 TI - Using trimethylamine dehydrogenase in an enzyme linked amperometric electrode. Part 1. Wild-type enzyme redox mediation. AB - An amperometric enzyme electrode was studied based on the wild-type protein trimethylamine dehydrogenase (TMADH), which catalyses the oxidative N demethylation of trimethylamine to produce dimethylamine and formaldehyde. Ferrocene derivatives were investigated electrochemically, as free diffusing electron acceptors for recycling of the prosthetic groups of the immobilised enzyme. Ferricinium had the highest rates but, inhibited the enzyme, possibly as a result of a conformational change initiated at the Val-344 residue where it binds close to the 4Fe-4S cluster, interrupting the electron transfer between flavin mononucleotide (FMN) and 4Fe-4S by changing the redox potential of one or both of the prosthetic groups. (Dimethylamino)methylene ferrocene (DMAMFe) (k(s) = 0.93 x 10(5) M(-1) s(-1)) did not show inhibition and was used as a comparison for steady-state characterisation. The sensor response was studied over the pH range 6.0-1.0. Plots of kcat/KM revealed two ionisations with pKa values of 7.5 and 10. The pKa of 10 was attributed to the ionisation of the secondary amine in DMAMFe, whereas the pKa of 7.5 was thought to reflect the ionisations of the intramolecular electron pathway. A TMADH/DMAMFe amperometric enzyme electrode was successfully used for the determination of TMA in different fish samples (detection limit: 2 mg TMA-N per lOOg wet fish muscle). The obtained results compared well with a reference method based on picric acid. PMID- 12625559 TI - Amperometric determination of sulfide at a pre-oxidised nickel electrode in acidic media. AB - The electrochemical response of a pre-oxidised nickel electrode to increasing additions of sulfide has been examined and shown to produce a stripping-like voltammetric wave. A mechanism is described based on the formation of nickel sulfide at the electrode surface from a non-electroactive nickel oxide layer. The analytical utility of the approach has been examined and a linear range from 1 to 140 microM and a limit of detection of 0.8 microM is achievable, depending on the accumulation time. PMID- 12625560 TI - Indirect voltammetric determination of trace hydroxylamine using magnetic microspheres. AB - A new indirect voltammetric method for the determination of hydroxylamine is described. It is based on the reduction of an electroactive derivative of hydroxylamine on the surface of a magnetic electrode. The electroactive derivative produced by hydroxylamine reacted with magnetic polymer microspheres containing carbonyl groups on the surface. The experimental conditions are discussed. It was found that the peak potential (Ep) of the derivative was -0.46 V (vs. Ag/AgCl) under optimum conditions. Hydroxylamine could be determined in the range of 5-2,000 microg l(-1) with the detection limit of 2 microg l(-1) and relative standard deviation for the determination of 100 microg l(-1) hydroxylamine was 2.35%. Satisfactory results were obtained for the determination of hydroxylamine in aqueous medium. PMID- 12625561 TI - Electrochemical immunoassay at a 17beta-estradiol self-assembled monolayer electrode using a redox marker. AB - A simple electrochemical immunoassay was demonstrated using a 17beta-estradiol modified electrode. 17beta-estradiol was immobilized on the gold electrode surface with a self-assembly technique. The specific binding between estradiol antibody and 17beta-estradiol on the electrode surface was evaluated by monitoring the change in the electrode response with three hydrophilic redox markers. The decrease in the electrode response for the redox marker was observed, when the antibody was bound to the estradiol self-assembled monolayer (SAM) electrode surface. The change in the electrode response of the redox marker is attributed to the steric hindrance between the antibody on the electrode surface and the redox marker. The relative standard deviation at 30 microg ml(-1) estradiol antibody was 4.1% (n = 3). The competitive reaction between the antigen in the solution and 17beta-estradiol immobilized on the electrode surface for the limited binding sites on the antibody produced an increase in the electrode response with hydroquinone as the marker. The binding affinity of three antigens including 17beta-estradiol to the estradiol antibody was evaluated. Furthermore, the result obtained from this method was compared with the previously reported enzyme binding assay using the biotinylated estradiol and the biotin-immobilized microtiter plate. PMID- 12625562 TI - Micro-Raman spectroscopy of natural and synthetic indigo samples. AB - In this work indigo samples from three different sources are studied by using Raman spectroscopy: the synthetic pigment and pigments from the woad (Isatis tinctoria) and the indigo plant (Indigofera tinctoria). 21 samples were obtained from 8 suppliers; for each sample 5 Raman spectra were recorded and used for further chemometrical analysis. Principal components analysis (PCA) was performed as data reduction method before applying hierarchical cluster analysis. Linear discriminant analysis (LDA) was implemented as a non-hierarchical supervised pattern recognition method to build a classification model. In order to avoid broad-shaped interferences from the fluorescence background, the influence of 1st and 2nd derivatives on the classification was studied by using cross-validation. Although chemically identical, it is shown that Raman spectroscopy in combination with suitable chemometric methods has the potential to discriminate between synthetic and natural indigo samples. PMID- 12625563 TI - Development of a modified medium pressure microwave vapor-phase digestion method for difficult to digest organic samples. AB - A previously developed microwave heated vapor-phase digestion method for biological samples was modified to enable digestion of difficult to digest organic samples. Organic samples containing ca. 100 mg of organic carbon were digested using volume calibrated quartz inserts inside second generation type medium pressure microwave vessels. As digestion reagents, 98% sulfuric acid, 70% nitric acid and 30% hydrogen peroxide were used. The accuracy of the method was tested with six certified reference materials. Elements Ca, Fe, K, Na, Mg, P and Zn were determined from NIST-SRM 8433 corn bran. Elements Al, Fe, Cd, Cu, and Zn were determined from NRCC DOLT-2 dogfish liver. The element Cd was determined from IRMM-VDA Cd in polyethylene No. 001-004 reference materials. These elements were determined from digested samples by ICP-OES. The results were close or within certified limits. The modified method could digest nearly all the materials tested, including the above mentioned reference materials, 2 nitrobenzoic acid (2-NBA), 4-NBA and copper(II) phthalosyanine-3, 4',4',4' tetrasulfonic acid tetrasodium salt (CPS). The method could not digest 3-NBA. PMID- 12625564 TI - Adsorption of water vapour from humid air in carbon molecular sieves: carbosieve S-III and carboxens 569, 1000 and 1001. AB - The adsorption of water vapour in carbon molecular sieves (CMS) used to determine volatile organic compounds (VOCs) in air was investigated. The CMS mass in the trap was found not to affect the mass of retained water under conditions of incomplete saturation of the adsorbent bed with water. Thus, the restrictions commonly imposed on the CMS mass are not necessary. The usefulness of four different CMSs to sample large volumes of humid air was estimated. Carboxen 1000 exhibited the best performance. To assess the magnitude of CMS mass in the trap in dependence on the volume, the relative humidity and the temperature of the sample, the use of a novel parameter, called the water vapour interference factor, was suggested. PMID- 12625565 TI - Quality of life in head and neck cancer. PMID- 12625566 TI - Neuroprotection of vestibular sensory cells from gentamicin ototoxicity obtained using nitric oxide synthase inhibitors, reactive oxygen species scavengers, brain derived neurotrophic factors and calpain inhibitors. AB - OBJECTIVE: In order to devise a new treatment for inner ear disorders, the efficacy of a nitric oxide synthase inhibitor (L-N(G)-nitroarginine methylester [L-NAME]), a radical scavenger (D-methionine), a neurotrophin (brain-derived neurotrophic factor [BDNF]) and a calpain inhibitor (leupeptin) for protection from hair cell damage was investigated. MATERIAL AND METHODS: The effects of these drugs on gentamicin-induced production of nitric oxide (NO) and reactive oxygen species (ROS) were studied by means of the fluorescence indicators 4,5 diaminofluorescein diacetate and dihydrotetramethylrosamine. The effect on gentamicin-induced vestibular hair cell damage was examined by using an in vitro LIVE/DEAD system. RESULTS: L-NAME inhibited the production of NO, D-methionine and BDNF restricted the production of ROS and leupeptin inhibited neither NO nor ROS. All the drugs used limited the vestibular hair cell damage caused by gentamicin. The combinations L-NAME + BDNF, L-NAME + leupeptin and D-methionine + BDNF had a significantly stronger preventive effect on hair cell damage. CONCLUSION: It is suggested that combined treatment with a radical inhibitor and either a neurotrophin or calpain inhibitor may help to treat inner ear disorders more effectively. PMID- 12625567 TI - Protective effect of brain-derived neurotrophic factor against the ototoxicity of Pseudomonas aeruginosa exotoxin A. AB - OBJECTIVE: The protective effect of brain-derived neurotrophic factor (BDNF) against the ototoxicity resulting from exposure of Pseudomonas aeruginosa exotoxin A (PaExoA) to the middle ear was analyzed. The combined effect of BDNF and N(G)-nitro-L-arginine methyl ester (L-NAME) was also investigated. MATERIAL AND METHODS: Six groups of albino rats were instilled through the tympanic membrane into the round window niche with the following solutions: saline; PaExoA; BDNF; L-NAME; PaExoA + BDNF; and PaExoA + BDNF + L-NAME. Frequency specific (2-31.5 kHz) auditory brainstem responses were used to obtain the hearing thresholds before and 2, 5 and 15 days after instillation. RESULTS: PaExoA penetrated from the middle ear into the cochlea, causing initially mixed hearing loss, followed by persistent sensorineural hearing loss. This impairment was blocked by BDNF at 6, 8 and 10 kHz on Day 2 and at 8 kHz on Day 5. L-NAME given in combination with BDNF did not show any additional protective effect. There were no significant differences in the thickness of the round window membrane between control ears and those in each instillation group. CONCLUSION: Our results suggest that BDNF may protect against cochlear damage caused by PaExoA in the middle turns of the ear. PMID- 12625569 TI - Immunological responses in acute low-tone sensorineural hearing loss and Meniere's disease. AB - OBJECTIVE: The autoimmune response appears to play an important role in some types of acute sensorineural hearing loss. Endolymphatic hydrops associated with fluctuating hearing loss has also been suggested to be caused by an immunological mechanism. Acute low-tone hearing loss (ALHL) associated with Meniere's disease (MD) is characterized by fluctuating hearing loss, and its etiology is thought to involve endolymphatic hydrops. The aim of this study was to attempt to determine the etiology of ALHL in MD. MATERIAL AND METHODS: A flow cytometer was used to analyze intracellular cytokine levels in peripheral blood from 19 patients with ALHL and 26 patients with MD and the data compared to those obtained from age- and gender-matched healthy volunteers. RESULTS: The patients with ALHL showed significantly increased levels of Th1 subsets (interferon-gamma-producing helper T cells) as compared to those in normal controls. The levels of Th2 (IL-4 producing helper T cells) subsets did not differ from those in the control group and thus Th1 predominated in ALHL patients. The patients with MD showed significantly increased natural killer cell activity but no Th1 dominance. These patients had no obvious systemic or local disease except in the inner ear. CONCLUSION: An abnormality of the Th1/Th2 balance in ALHL and increased natural killer cell activity in MD are thought to relate to inner ear disorder. These results are consistent with the possibility that the etiology of ALHL and MD involves an immune response. PMID- 12625570 TI - Head and body center of gravity control strategies: adaptations following vestibular rehabilitation. AB - OBJECTIVE: We present for the first time evidence that vestibulopathy impairs coordination of the head with the body center of gravity (CG) during free speed gait over ground. Vestibulopathic individuals demonstrate uncoordinated movement and gait due, at least in part, to impaired head stability and visual fixation. Vestibular rehabilitation increases speed and stability during gait and stair climbing, although the underlying mechanisms are poorly understood. MATERIAL AND METHODS: To determine whether these locomotor improvements are due to reorganized coordination of the head with whole body CG, three-dimensional kinematics were obtained from 10 vestibulopathic individuals before and after vestibular rehabilitation and from 10 matched healthy control subjects during unconstrained, paced and in-place gait. Head control patterns were characterized using both qualitative pattern analysis and quantification of coherence between head and body CG displacements. RESULTS: Patterns of head-CG coordination differ between normal and vestibulopathic individuals in all three directions of head rotation- pitch, roll and yaw--before rehabilitation. Following vestibular rehabilitation, subjects with vestibulopathy demonstrate more normal patterns in pitch and improvements toward normal in roll and yaw. CONCLUSION: These data strongly suggest that compensatory mechanisms, obtained during vestibular rehabilitation, mediate head-CG coordination. PMID- 12625568 TI - Clostridium difficile toxin B, an inhibitor of the small GTPases Rho, Rac and Cdc42, influences spiral ganglion neurite outgrowth. AB - OBJECTIVE: Neurotrophins and extracellular matrix (ECM) molecules are involved in neurite guidance during the development of spiral ganglion (SG) neurons. Several intracellular signaling molecules can be activated by ECMs and neurotrophins via their cognate receptors. In other systems these include the Rho small GTPases, which influence reorganization of the actin cytoskeleton that is required for axon growth. The aim of this study was to determine whether neurotrophin-3 (NT-3) mediated SG neurite outgrowth on laminin-1 (LN) is dependent on the activation of the small GTPases Rho/Rac/Cdc42. MATERIAL AND METHODS: SG explants from postnatal day 4 rats were cultured on LN with and without NT-3 and increasing concentrations of Clostridium difficile Toxin B, an inhibitor of Rho GTPases. After fixation and immunocytochemical labeling, neurite growth was evaluated. RESULTS: Treatment with C. difficile Toxin B without NT-3 led to a dose-dependent decrease in the length and number of processes on LN. In contrast, C. difficile Toxin B had no significant effect on NT-3-mediated stimulation of neurite growth on LN, in terms of either number or length. CONCLUSION: The results suggest that the Rho GTPases (Rho, Rac and Cdc42) are not involved in the pathways linking NT 3 signals to neurite outgrowth, but appear to be involved in LN signaling in these neurons. However, NT-3 can override or bypass LN signaling to promote neurite extension. PMID- 12625571 TI - Influenza A virus infection of human Schwann cells in vitro. AB - OBJECTIVES: Sudden sensorineural hearing loss, vestibular neuronitis, vocal fold paralysis and Bell's palsy have been associated with a viral etiology, due to the infection of nerve cells. The goal of this research was to ascertain whether Schwann cells can support infection with human influenza A virus and thereby represent a plausible alternative site for virus-host interaction. Viral infection of Schwann cells may lead to secretion of inflammatory mediators, leukocyte recruitment, demyelination and nerve damage. MATERIAL AND METHODS: Cultured human Schwann cells were exposed to human influenza A virus. Infection was assayed at various times post-inoculation (0, 24, 48 and 72 h) using light microscopy, immunocytochemistry and influenza A virus-specific reverse transcriptase polymerase chain reaction (RT-PCR). A group of unexposed cells served as controls. RESULTS: Following exposure to the virus, vacuolization, cellular expansion and detachment from the dish were seen as early as 24 h post inoculation. The exposed cells demonstrated positive immunocytochemical staining for influenza A virus antigen at 24, 48 and 72 h. Using RT-PCR, a sharp rise in influenza A virus-specific mRNA was detected. CONCLUSIONS: Human Schwann cells can be infected with human influenza A virus. Further studies will assess the inflammatory response in this model. PMID- 12625573 TI - Hearing deterioration in patients with a non-growing vestibular schwannoma. AB - OBJECTIVE: The aim of this study was to document and analyse the course of several audiometric parameters in 49 patients with a non-growing unilateral vestibular schwannoma (VS). MATERIAL AND METHODS: Patients received conservative management and absence of tumour growth was ascertained by means of serial magnetic imaging studies. Pure-tone audiometry and speech audiometry were performed at yearly intervals. RESULTS: Pure-tone audiometry revealed a significant increase in thresholds at all frequencies, except for 8.0 kHz. The maximum yearly threshold increase was 2.4 dB hearing level at 1.0 and 2.0 kHz. Speech audiometry revealed a significant decrease in maximum discrimination over the course of time. No significant changes were observed in the following parameters: the intensity level at which maximum discrimination was achieved; the roll-over index; the speech reception threshold; and the slope of the curve in the speech audiogram. No change was observed in the relation between pure-tone audiometry thresholds and speech audiometry scores. Apparently, the deterioration of pure-tone perception and speech discrimination ran parallel courses. CONCLUSIONS: The results of this study indicate that hearing loss is a predominant symptom in patients with a non-growing VS, as is also known in patients with a growing lesion. Moreover, it seems unlikely that the hearing loss in VS patients is merely the result of mechanical influences on retrocochlear neural or vascular structures. PMID- 12625572 TI - Defibrinogenation therapy for idiopathic sudden sensorineural hearing loss in comparison with high-dose steroid therapy. AB - OBJECTIVE: The efficacy of defibrinogenation therapy for idiopathic sudden sensorineural hearing loss was studied in comparison with high-dose steroid therapy. MATERIAL AND METHODS: Eighty-eight consecutive patients with hearing levels > 40 dB and who had suffered hearing loss for < or = 30 days were enrolled: 40 patients for high-dose steroid therapy (PSL group) and 48 for defibrinogenation therapy (BX group). Hearing recovery was evaluated by grade assessment and by the improvement in hearing compared to the unaffected contralateral ear. RESULTS: The overall hearing outcomes of the two groups were roughly equivalent. However, with regard to patients with initial hearing levels < 80 dB, the hearing improvement rate of the BX group was significantly worse than that of the PSL group (61.2% +/- 7.3% vs 88.7% +/- 8.9%; p < 0.05), whereas in patients with initial hearing levels > or = 80 dB, the hearing outcomes did not differ between the 2 groups. Three patients in the PSL group manifested hyperglycemia while no serious side-effects were observed in the BX group. CONCLUSION: These results indicate that high-dose steroid therapy should be employed in preference to defibrinogenation therapy for patients with moderate hearing loss, whereas defibrinogenation therapy has an advantage for those with severe hearing loss, in view of its lower frequency of side-effects. PMID- 12625574 TI - Cochlear implantation in a patient with mitochondrial encephalopathy, lactic acidosis and stroke-like episodes syndrome. AB - OBJECTIVE: The aim of this study was to investigate the usefulness of a Nucleus C124M cochlear implant in a patient with mitochondrial encephalopathy, lactic acidosis and stroke-like episodes (MELAS) syndrome. MATERIAL AND METHODS: A 29 year-old woman with MELAS syndrome presented with sensorineural hearing loss and diabetes mellitus and underwent multichannel cochlear implantation. The follow-up period was 10 months. The case history, assessment of mitochondrial disease, indications for the cochlear implant and the benefits of cochlear implantation were evaluated. RESULTS: Nine months after the surgery the patient could use the telephone and was quite satisfied with the improvement in communication due to the cochlear implant. CONCLUSION: Cochlear implantation can be recommended for patients with MELAS syndrome if they have residual retrocochlear function. Single photon emission computerized tomography was found to be very useful for evaluating retrocochlear function. PMID- 12625575 TI - Neurotological complications after radiosurgery versus conservative management in acoustic neuromas: a systematic review-based study. AB - OBJECTIVE: Treatment modalities for acoustic neuroma (AN) include surgery, observation and gamma-knife surgery. The aim of this study was to compare neurotological complications resulting from two treatment alternatives to microsurgery: radiosurgery and observation. MATERIAL AND METHODS: We conducted a systematic review of the literature dealing with radiosurgery for AN and compared the rate of neurotological complications in this population with that in a cohort of patients managed conservatively. RESULTS: We found that neurotological complications, namely facial hypoesthesia (p = 0.002), hearing loss (p < 0.05) and hydrocephalus (p = 0.02), were more frequently encountered after radiosurgery than with conservative management. In contrast, we found that the risk of growth of AN is significantly higher with conservative management and that the rate of stability of the tumor did not differ significantly between the two treatments. CONCLUSION: We prefer a conservative management regimen for patients who cannot be operated on for their AN. However, there are some difficulties inherent in this conservative management policy, namely non-compliance and difficulties in establishing the evolution of the tumor. PMID- 12625576 TI - The reversed ipsilateral acoustic reflex: clinical features and kinetic analysis. AB - OBJECTIVE: The reversed ipsilateral acoustic reflex (RIAR) has been the subject of few studies in the past literature and the nature of this "reflex" response is not yet completely understood. It can be observed in different diseases, all of which result from inactivity of the stapedial reflex (SR). We aimed to investigate the occurrence of the RIAR in different diseases. MATERIAL AND METHODS: An extensive clinical investigation of the RIAR was carried out. RESULTS: The RIAR was found to be present in 100% of ears lacking an effective SR arc. Its amplitude is modulated by the air pressure level in the external ear canal whilst its sign is constantly positive and never reverses. CONCLUSION: Kinetic analysis of the RIAR suggests the existence of a single underlying phenomenon while the interaction of the RIAR with the SR may explain the complex morphology frequently observed in the normal ipsilateral acoustic reflex. PMID- 12625577 TI - Solitary fibrous tumour of the nasal cavity and paranasal sinuses. AB - OBJECTIVE: Solitary fibrous tumour (SFT) is an uncommon unilateral benign lesion that usually arises from the pleura. Recently, SFT has been described in many other locations, such as the mediastinum, head and neck, orbit and urogenital system. To date, only 21 cases of SFT arising from the nasal cavity and paranasal sinuses have been reported in the literature. MATERIAL AND METHODS: We describe a case of SFT localized in the right nasal cavity with extension to the right ethmoid and sphenoid sinuses in a 43-year-old man who presented with a 6-month history of unilateral right-sided nasal obstruction, rhinorrhoea and recurrent epistaxis. RESULTS: CT showed a large mass arising from the right anterior ethmoid and sphenoid sinuses. Histopathologically, the lesion consisted of a non encapsulated mass with spindle-shaped cells within a collagenous stroma. Definitive diagnosis was obtained by means of immunohistochemical analysis, which showed vimentin and CD34 cells. CONCLUSION: The tumour was removed by endoscopic sinus surgery, and the patient remained free of tumour after 12 months of follow up. PMID- 12625578 TI - Prevalence of self-reported allergic and non-allergic rhinitis symptoms in Stockholm: relation to age, gender, olfactory sense and smoking. AB - OBJECTIVE: To estimate the prevalence of isolated self-reported allergic and non allergic rhinitis symptoms in an adult population and to explore the relations to age, gender, olfaction and smoking habits. MATERIAL AND METHODS: Self-judged health and environmental exposures were investigated by means of a questionnaire survey administered to a stratified random sample of 15,000 adults in Stockholm County. RESULTS: A total of 10,670 individuals were included in the analysis, corresponding to a response rate of 73%. The results revealed a high prevalence of self-reported non-allergic rhinitis, 19%, almost as high as the prevalence of self-reported allergic rhinoconjunctivitis, 24%. In contrast to current clinical opinion, we did not find a significant increase in the prevalence of non-allergic symptoms with increased age. There were no statistically significant gender differences in the prevalence of either allergic or non-allergic symptoms. A reduced sense of smell was twice as common in the non-allergic group, 23%, as in the healthy population. The prevalence of rhinitis symptoms differed according to smoking habits. CONCLUSION: Both self-reported allergic rhinitis symptoms and non allergic nasal symptoms are frequent in the population sample. Self-reported non allergic nasal symptoms seem to occur independent of age and reduced olfactory sense is a common complaint among these subjects. The prevalence of self-reported allergic and non-allergic nasal symptoms did not differ much between men and women or between individuals with different smoking habits. PMID- 12625579 TI - Antimicrobial peptide LL-37 is upregulated in chronic nasal inflammatory disease. AB - OBJECTIVE: LL-37 is one of the antimicrobial peptides and the only member of the cathelicidin family identified so far in humans. We attempted to find a correlation between LL-37 peptide and inflammation of the nasal mucosa. MATERIAL AND METHODS: Nasal mucosa specimens were obtained from 15 chronic infective rhinitis patients and 6 normal controls. Immunohistochemical staining was used to examine the localization of LL-37 and reverse transcriptase polymerase chain reaction (RT-PCR) was used to determine the levels of LL-37, IL-1beta and IL-8 in tissue. RESULTS: LL-37 peptide was primarily localized in the surface of the epithelia, in the serous and mucous cells of the submucosal glands and in stromal inflammatory cells. The number of LL-37 immunoreactive cells in inflammatory nasal mucosa was significantly increased compared with normal nasal tissue. Using RT-PCR, LL-37 mRNA was detected in 3/6 normal turbinate samples but in all cases with inflammatory nasal tissues. IL-1beta and IL-8 transcripts exhibited a similar pattern to that of LL-37. CONCLUSIONS: We suggest that LL-37 is one of the antimicrobial peptides found in human nasal mucosa and that it participates in the innate immune system of the nasal mucosa. PMID- 12625580 TI - Spindle-cell tumours of the larynx: diagnostic pitfalls. A case report and review of the literature. AB - Malignant laryngeal tumours with spindle-cell morphology are relatively rare. Differential diagnosis of spindle-cell carcinoma, leiomyosarcoma, fibrosarcoma, rhabdomyosarcoma, malignant melanoma and inflammatory myofibroblastic tumour may be particularly difficult. Ambiguous and indeterminate diagnoses are also possible after immunohistochemical investigations of the expression of specific tumour markers but electron microscopic observations help to clarify most of these diagnoses. PMID- 12625581 TI - Development and clinical assessment of a heat and moisture exchanger with a multi magnet automatic tracheostoma valve (Provox FreeHands HME) for vocal and pulmonary rehabilitation after total laryngectomy. AB - OBJECTIVE: To develop and test the prototypes of a novel post-laryngectomy rehabilitation tool incorporating an obligatory, disposable heat and moisture exchanger (HME) and a reusable, multi-magnet automatic speaking valve (ASV). MATERIAL AND METHODS: The study subjects comprised 20 laryngectomized individuals (15 males, 5 females), 5 of whom were already using an ASV and 15 who were not. Three successive prototypes were tested. Data were collected by means of structured questionnaires, considering for example patient compliance, skin adhesion, voicing and coughing aspects, and voice and speech quality assessments, assessing for example maximum phonation time and dynamic loudness range. RESULTS: Of the 15 non-ASV users, 5 did not comply with the study due to peristomal skin adhesion problems. Of the remaining 15 patients, all 5 ASV users and 6/10 non users were fully compliant with the new device. The cough-relief valve of the new device functions properly, as does the valve position adjustment for physical exertion. With this new device the maximum phonation time was longer than with a regular ASV (15.2 vs 11.6 s; p = 0.006) and the dynamic range was larger (33.0 vs 24.8 dB; p < 0.001). CONCLUSION: The test results obtained with this new device show that its advanced features (obligatory HME and multi-magnet valve systems) offer additional benefits for further improving vocal and pulmonary rehabilitation after total laryngectomy. PMID- 12625582 TI - Potential molecular prognostic markers for lymph node metastasis in head and neck squamous cell carcinoma. PMID- 12625583 TI - Vitamin A-storing stellate cells in the human vocal fold. AB - OBJECTIVE: Previously, we discovered cells with a star-like appearance that constantly synthesized extracellular matrices in the human adult vocal fold mucosa. These cells had no nomenclature and were thus designated "vocal fold stellate cells (VFSC)". The aim of the present study was to detect vitamin A (retinol) in the VFSC. MATERIAL AND METHODS: Five normal human adult vocal folds obtained from surgical specimens were used. Light microscopy using the gold chloride method and fluorescence microscopy were used to detect vitamin A. RESULTS: VFSC distributed in the maculae flavae of human adult vocal folds possessed lipid droplets and stored vitamin A. CONCLUSION: It is generally accepted that vitamin A, an essential liposoluble vitamin, is used as a morphogen that controls the differentiation and morphogenesis of cells. In our previous investigation and in this study VFSC were found to differ morphologically from conventional fibroblasts. VFSC represent morphologically a new category of cells in the human vocal fold. PMID- 12625584 TI - Measurement of longitudinal piezoelectric coefficient of thin films by a laser scanning vibrometer. AB - A laser scanning vibrometer (LSV) was used for the first time to measure the piezoelectric coefficient of ferroelectric thin films based on the converse piezoelectric effect. The significant advantages of the use of the LSV for this purpose were demonstrated. Several key points were discussed in order to achieve reliable and accurate results. PMID- 12625585 TI - Stability analysis of an acoustically levitated disk. AB - In this paper, a model is developed for the stability analysis of an acoustically levitated disk on the basis of analyzing eddy acoustic streaming and acoustic viscous stress. In the model, the effect of the acoustic streaming outside the boundary layer that is on the surface of the levitated disk is properly taken into account. Also, the calculation of sound field and acoustic viscous stress is limited to the range that has a dominant effect on the stability. By this method, we obtain a quite accurate solution of the stability coefficient. For the small horizontal shift of a large levitated disk, the model is verified by the good agreement between the experimental and theoretical results. By means of this model and relevant experiments, some factors that affect the stability of the levitated disk are investigated, and useful guidelines for design and application are obtained. It is found that the range from the edge to the outermost nodal circle of the disk-shaped vibrator has a large effect on the stability of the levitated disk. To stabilize the levitated disk by acoustic viscous force, the distance between the edge and the outermost nodal circle of the vibrator must be larger than a critical value, which is determined by the driving frequency and the sound velocity of the fluid between the levitated disk and the vibrator. When this condition is satisfied, increasing the distance between the edge and the outermost nodal circle leads to a decrease in the stability. It is also found that the property of the fluid between the levitated disk and the vibrator has a large effect on the stability. It is easier to stabilize the levitated disk in steam than in air, but more difficult to do so in carbon dioxide and hydrogen. In addition, theoretical results show that increasing the weight per unit area of the levitated object increases the stability for a given vibrator velocity. The distribution of the acoustic viscous stress and the dependence of the stability coefficient and the holding force on the horizontal shift of the levitated disk, which are obtained by this study, also are useful to a better understanding of the stability of the levitated disk. PMID- 12625586 TI - Adaptive imaging using the generalized coherence factor. AB - Sound-velocity inhomogeneities degrade both spatial and contrast resolutions. This paper proposes a new adaptive imaging technique that uses the generalized coherence factor (GCF) to reduce the focusing errors resulting from the sound velocity inhomogeneities. The GCF is derived from the spatial spectrum of the received aperture data after proper receive delays have been applied. It is defined as the ratio of the spectral energy within a prespecified low-frequency range to the total energy. It is demonstrated that the low-frequency component of the spectrum corresponds to the coherent portion of the received data, and that the high-frequency component corresponds to the incoherent portion. Hence, the GCF reduces to the coherence factor defined in the literature if the prespecified low-frequency range is restricted to dc only. In addition, the GCF is also an index of the focusing quality and can be used as a weighting factor for the reconstructed image. The efficacy of the GCF technique is demonstrated for focusing errors resulting from the sound-velocity inhomogeneities. Simulations and real ultrasound data are used to evaluate the efficacy of the proposed GCF technique. The characteristics of the GCF, including the effects of the signal-to noise ratio and the number of channels, are also discussed. The GCF technique also is compared with the correlation-based technique and the parallel adaptive receive compensation algorithm; the improvement in image quality obtained with the proposed technique rivals that of the latter technique. In the presence of a displaced phase screen, this proposed technique also outperforms the correlation based technique. Computational complexity and implementation issues also are addressed. PMID- 12625587 TI - Dynamic-transmit focusing using time-dependent focal zone and center frequency. AB - A method for achieving dynamic-transmit focus is presented. Within an initial high bandwidth pulse, successively higher frequencies are focused to successively closer focal zones along a transmitted beam line. During the receive operation, a time varying bandpass filter initially passes the higher frequencies that are focused closer in and successively passes lower center frequencies as time evolves and the focal zone moves out. The dynamically focused receive pulses are digitally sampled and processed by a matched digital filter to minimize phase anomalies. In this way, an improved resolution image is obtained with no loss of frame rate. The method is evaluated using comprehensive simulations that account for realistic levels of phase aberration and tissue attenuation. The method is relatively robust with respect to these perturbations. When the appropriate conditions apply, the new method can achieve an improvement in mean lateral resolution similar to that found in a multiple transmit zone implementation but without the frame rate penalty. A discussion of implementation considerations and limitations is presented. PMID- 12625588 TI - Superthreshold behavior and threshold estimation of ultrasound-induced lung hemorrhage in pigs: role of age dependency. AB - Age-dependent threshold and superthreshold behavior of ultrasound-induced lung hemorrhage were investigated with 116 2.1 +/- 0.3-kg neonate crossbred pigs (4.9 +/- 1.6 days old), 103 10 +/- 1.1-kg crossbred pigs (39 +/-5 days old), and 104 20+/-1.2-kg crossbred pigs (58 +/- 5 days old). Exposure conditions were: 3.1 MHz, 10-s exposure duration, 1-kHz pulse repetition frequency (PRF), and 1.2 micros pulse duration. The in situ (at the pleural surface) peak rarefactional pressure ranged between 2.2 and 10.4 MPa with either eight or nine acoustic pressure groups for each of the three pig ages (12 pigs/exposure group) plus sham exposed pigs. There were no lesions in the shams. Pigs were exposed bilaterally with the order of exposure (left then right lung, or right then left lung) and acoustic pressure both randomized. Pig age was not randomized. Individuals involved in animal handling, exposure, and lesion scoring were blinded to the exposure condition. Logistic regression analysis was used to examine the dependence of the lesion incidence rates on in situ peak rarefactional pressure, left versus right lung exposure, order of exposure (first versus second), and age in three age groups. Likewise, lesion depth and lesion root surface area were analyzed using Gaussian tobit regression analysis. A significant threshold effect on lesion occurrence was observed as a function of age; younger pigs were less susceptible to lung damage given equivalent in situ exposure. Overall, the oldest pigs had a significantly lower threshold (2.87 +/- 0.29 MPa) than middle-aged pigs (5.83 +/- 0.52 MPa). The oldest pigs also had a lower threshold than neonate pigs (3.60 +/- 0.44 MPa). Also, an unexpected result was observed. The ultrasound exposures were bilateral, and the threshold results reported above were based on the lung that was first exposed. After the first lung was exposed, the pig was turned over and the other lung was exposed to the same acoustic pressure. There was a significant decrease (greater than the confidence limits) in occurrence thresholds: 3.60 to 2.68, 5.83 to 2.97, and 2.87 to 1.16 MPa for neonates, middle aged, and oldest pigs, respectively, in the second lung exposed. Thus, a subtle effect in lung physiology resulted in a major effect on lesion thresholds. PMID- 12625589 TI - Optimization of buffer rod geometry for ultrasonic sensors with reference path. AB - Several applications of ultrasonic techniques are limited by the signal-to-noise ratio (SNR). Transducers in these applications usually operate in the pulse-echo mode. Many transducers, especially those for high temperatures, use buffer rods. Often a reference path is used to eliminate electrical and transducer drift. Interference of echo signals and noise causes errors of both amplitude and phase measurement of the detected echoes. In this paper we discuss the influence of major noise sources as a function of geometry and operating environment. The effects are studied using both experimental results and models. Although the results are applied to an ultrasonic density sensor operating in the pulse-echo mode, they are applicable to other pulse-echo mode transducers comprising homogeneous cylindrical buffer rods. This paper will show how the SNR of the density transducer was improved in a special time window from 34 to 72 dB by careful design. PMID- 12625590 TI - A single-element tuning fork piezoelectric linear actuator. AB - This paper describes the design of a piezoelectric tuning-fork, dual-mode motor. The motor uses a single multilayer piezoelectric element in combination with tuning fork and shearing motion to form an actuator using a single drive signal. Finite-element analysis was used in the design of the motor, and the process is described along with the selection of the device's materials and its performance. Swaging was used to mount the multilayer piezoelectric element within the stator. Prototypes of the 25-mm long bidirectional actuator achieved a maximum linear no load speed of 16.5 cm/s, a maximum linear force of 1.86 N, and maximum efficiency of 18.9%. PMID- 12625591 TI - Equations for small fields superposed on finite biasing fields in a thermoelectroelastic body. AB - Equations for infinitesimal incremental fields superposed on finite biasing fields in a thermoelectroelastic body are derived from the nonlinear equations of thermoelectroelasticity. The equations are general in the sense that no assumptions are made on the biasing fields. The general equations obtained are reduced to various special cases under different approximations. PMID- 12625592 TI - Conventional detection of 2,4-dinitrophenol using quartz crystal microbalance. AB - We present conventional detection of 2,4-dinitrophenol (DNP) for using the competitive reaction between DNP and DNP-conjugated albumin onto DNP antibody immobilized quartz crystal microbalance (QCM). This QCM method allows detection of DNP concentration in the range of 0.01 to 100 ng/ml; linear correlation obtains DNP concentration from 1 to 100 ng/ml. PMID- 12625593 TI - Contrast nephropathy or contrast creatininopathy. PMID- 12625594 TI - Matrix metalloproteinases and coronary artery diseases. AB - Matrix metalloproteinases (MMPs) play an important role in cardiovascular remodeling by degrading the extracellular matrix. Enhanced MMP expression has been detected in the atherosclerotic plaque, and activation of MMPs appears to be involved in the vulnerability of the plaque. Circulating MMP levels are elevated in patients with acute myocardial infarction and unstable angina. Increased MMP expression is also observed after coronary angioplasty, which is related to late loss index after the procedure. These observations suggest that MMP expression may be not only related to instability of the plaque, but also to the formation of restenotic lesions. The development of therapeutic drugs targeted specifically against MMPs may be useful in the prevention of atherosclerotic lesion development, plaque rupture, and restenosis. PMID- 12625595 TI - Comparison of left ventriculography and coronary arteriography with positron emission tomography in assessment of myocardial viability. AB - BACKGROUND: Assessment of viability of myocardium after an ischemic insult is an important clinical question that affects decisions pertaining to potential revascularization. The results of contrast left ventriculograms and coronary angiography were compared to positron emission tomography (PET) in 64 patients with coronary artery disease and reduced left ventricular function. HYPOTHESIS: The study was undertaken to determine the relative utility of the invasive studies in the assessment of viability. METHODS: Right anterior oblique ventriculograms were assessed for hypokinesis, akinesis, or dyskinesis in six segments. The PET scans were assessed for viability by visual estimation of flourodeoxyglucose (FDG) uptake in six segments that corresponded to the segments analyzed on the ventriculograms. RESULTS: Of a total of 373 segments successfully analyzed by PET, 272 were judged to be viable (normal or hypokinetic) by contrast ventriculography. Of these, 253 (93%) were considered viable by PET. Of 177 segments deemed either normal or mild-to-moderately hypokinetic by ventriculography, 170 (94%) were viable by PET. Of 95 severely hypokinetic segments, 83 (84%) were viable by PET. Of 79 akinetic segments, 44 (56%) were considered viable by PET. For segments that were dyskinetic and thought to be nonviable by ventriculography, 19 of 22 (86%) were also considered nonviable by PET. For 294 segments for which a determination on viability was made based on the presence of wall motion on the ventriculogram (normal, hypokinetic, ordyskinetic; not akinetic), there was excellent agreement with PET (93%; p < 0.001). In 49 patients there was akinesis in no more than one segment in either the anterior or inferior territories, indicating the potential for assessment of viability by ventriculography in at least two of three segments in each territory. Coronary anatomy was analyzed to assess whether coronary patency could help in assessing viability. Segments supplied by patent arteries were more likely to be viable by PET than segments supplied by occluded arteries (p < 0.001). Akinetic segments were more likely to be supplied by occluded arteries (56 vs. 23, 72%). Dyskinetic segments were predominantly nonviable by PET (86%) and were usually supplied by occluded arteries (77%). CONCLUSION: In patients in whom the assessment of viability is clinically relevant, the presence of systolic inward motion on the contrast left ventriculogram correlates well with segment viability by PET, while outward or dyskinetic movement correlates well with nonviability. Thus, the use of PET to assess viability in many patients may be unnecessary. PMID- 12625596 TI - Predischarge exercise electrocardiogram and stress echocardiography can predict long-term clinically driven revascularization following acute myocardial infarction. AB - BACKGROUND: Predischarge stress testing provides suboptimal prediction of spontaneous hard events following uncomplicated acute myocardial infarction (AMI). HYPOTHESIS: This study was aimed at assessing whether soft cardiac ischemic events requiring late revascularization could be predicted more accurately. METHODS: In all, 428 patients undergoing exercise electrocardiography (ECG) and stress echocardiography (SE, 345 dobutamine and 83 dypiridamole) within 15 days of uncomplicated AMI were followed up for 425 (range 20-2220) days. Soft ischemic events (effort angina>class II [Canadian Cardiovascular Society Classification] and unstable angina) driving late (>6 months) revascularization were regarded as endpoints. RESULTS: A total of 58 events (29 effort and 29 unstable angina with subsequent 47 coronary artery bypass grafts and 11 percutaneous transluminal coronary angioplasties) occurred: 26 in patients with positive exercise ECG and 34 in patients with positive SE. Univariate predictors of revascularizations were positive exercise ECG (p = 0.0001), peak wall motion score index (WMSI) (p = 0.0009), low workload (p = 0.0018), rest WMSI (p = 0.02) and positive SE (p = 0.02). Cox multivariate analysis selected peak WMSI, positive exercise ECG, and low workload positive exercise ECG as independent predictors of late revascularizations. CONCLUSIONS: Predischarge stress testing identifies the long-term occurrence of soft ischemic events driving late revascularization after uncomplicated AMI. PMID- 12625597 TI - Hemodynamic changes and prognosis in patients with hypertrophic cardiomyopathy and abnormal blood pressure responses during exercise. AB - BACKGROUND: An abnormal blood pressure response (BPR) during exercise has been proposed as a risk factor for sudden cardiac death in patients with hypertrophic cardiomyopathy (HCM). Some patients with HCM show systolic dysfunction during exercise. HYPOTHESIS: The aim of this study was to clarify the hemodynamic response during exercise and prognosis in patients with HCM and abnormal BPR. METHODS: Sixty-five patients with HCM underwent radionuclide monitoring of left ventricular function and measurement of blood pressure during supine ergometer exercise. Thereafter, cardiac events were recorded for an average period of 76 months. RESULTS: Seven of 65 patients had abnormal BPR, while the others had normal BPR. Changes of heart rate and systemic vascular resistance during exercise did not differ between the two groups. Stroke volume did not increase in the abnormal BPR group but did in the normal BPR group. During a mean follow-up period of 76 months, three of the seven patients (43%) with abnormal but only one patient (2%) with normal BPR suffered a malignant arrhythmia. CONCLUSIONS: Abnormal BPR occurred in about 11% of patients with nonobstructive HCM and was associated with a high prevalence of cardiac events. The predictor of abnormal BPR during exercise may not be an abnormal response of systemic vascular resistance and heart rate, but the lack of an appropriate increase in stroke volume. PMID- 12625598 TI - Characteristics and outcome of patients with ST-elevation infarction in relation to whether they received thrombolysis or underwent acute coronary angiography: are we selecting the right patients for coronary angiography? AB - BACKGROUND: During the last decade, there has been an on-going debate with regard to whether percutaneous coronary intervention (PCI) or thrombolysis should be preferred in patients with ST-elevation acute myocardial infarction (AMI). Some studies clearly advocate PCI, while others do not. HYPOTHESIS: The study aimed to describe the characteristics and to evaluate outcome of patients with suspected ST-elevation or left bundle-branch block infarction in relation to whether they received thrombolysis or had an acute coronary angiography aiming at angioplasty. METHODS: The study included all patients admitted to Sahlgrenska University Hospital in Goteborg, Sweden, with suspected acute myocardial infarction who, during 1995-1999, had ST-elevation or left bundle-branch block on admission electrocardiogram (ECG) requiring either thrombolysis or acute coronary angiography. A retrospective evaluation with a follow-up of 1 year after the intervention was made. RESULTS: In all, 413 patients had thrombolytic treatment and 400 had acute coronary angiography. The patients who received thrombolysis were older (mean age 70.3 vs. 64.1 years). Mortality during 1 year of follow-up was 20.9% in the thrombolysis group and 16.6% in the angiography group (p = 0.12). Among patients in whom acute coronary angiography was performed, only 85% underwent acute percutaneous coronary intervention (PCI). There was a mortality of 12.1 vs. 41.7% among those who did not undergo acute PCI. Development of reinfarction, stroke, and requirement of rehospitalization was similar regardless of type of initial intervention. The thrombolysis group more frequently required new coronary angiography (36.9 vs. 20.6%; p<0.0001) and new PCI (17.8 vs. 11.9%; p = 0.01). Despite this, after 1 year symptoms of angina pectoris were observed in 27% of patients in the thrombolysis group and in only 14% of those in the angiography group (p = 0.0002). CONCLUSION: In a Swedish university hospital with a high volume of coronary angioplasty procedures, we found no significant difference in mortality between patients who had thrombolysis and those who underwent acute coronary angiography. However, requirement of revascularization and symptoms of angina pectoris 1 year later was considerably less frequent in those who had undergone acute coronary angiography. However, distribution of baseline characteristics was skewed and efforts should be focused on the selection of patients for the different reperfusion strategies. PMID- 12625599 TI - C-reactive protein and rapidly progressive coronary artery disease--is there any relation? AB - BACKGROUND: High plasma C-reactive protein (CRP) levels have been associated with an unfavorable outcome in patients with coronary artery disease (CAD), and a direct participation of CRP in the atherosclerotic process has been postulated. HYPOTHESIS: The aim of this study was to evaluate the possible relationship of high plasma CRP levels with the rapid progression of coronary atherosclerosis (RPCAD). METHODS: In all, 194 patients who were readmitted and underwent repeat coronary angiography because of recurrence of symptoms following successful percutaneous coronary intervention were studied. Median angiographic follow-up time was 6 months. Rapid progression CAD was defined as the presence of a new lesion, > 25% in luminal diameter stenosis, in a previously nondiseased vessel, or deterioration of a known, nontreated lesion by at least 25%. RESULTS: By multivariate analysis, patients with high plasma CRP levels upon first admission were at higher risk of RPCAD. In particular, odds ration (OR) = 1.8; 95% confidence interval (CI) = 1.3-3.6; p value = 0.02 in patients with CRP = 0.5-2 mg/dl versus patients with CRP < 0.5 mg/dl, and OR = 7.1; 95% CI = 3.8-9.5; p value < 0.001 in patients with CRP > 2 mg/dl versus patients with CRP < 0.5 mg/dl. CONCLUSION: Increased plasma CRP levels could possibly identify patients at high risk for the development of RPCAD. PMID- 12625600 TI - Systolic, diastolic, and pulse pressures as coronary risk factors in a population with low cholesterol levels: a prospective 10-year evaluation. AB - BACKGROUND: Pulse pressure (PP) has recently been found to be predictive of subsequent cardiovascular, particularly coronary, events in some middle-aged and elderly populations. HYPOTHESIS: The aim of this prospective population-based study was to evaluate the roles of systolic (SBP), diastolic blood pressure (DBP), and PP in predicting coronary heart disease (CHD) morbidity and mortality in both genders in a population of a wide age span and a setting of low levels of low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesterol. METHODS: Of participants in the Turkish Adult Risk Factor Study aged > or = 20 years, those free of CHD at baseline examination in 1990 were followed up for a mean of 9.3 years. Coronary heart disease was diagnosed based on clinical findings and Minnesota coding of resting electrocardiograms (ECGs). In 2,601 men and women (mean age at baseline 41.4 +/- 15 years), deaths from CHD developed in 80 and nonfatal CHD in 158 patients. RESULTS: In a logistic regression analysis for predictors of composite endpoint of coronary mortality and morbidity, individually. DBP was not significantly and independently associated, while SBP was an independent risk predictor in both genders (relative risk [RR] 1.016). When two of three blood pressure components were entered jointly into the multivariable model, PP had a value inferior to that of SBP among men and women, but was of greater value than DBP in men in predicting both the composite endpoint and death from CHD. Among women, the predictive values of PP and DBP were similar. Within the categories of SBP > or = 120 mmHg in men, PP contributed significantly to the predictive power of SBP in overall CHD risk over and above that supplied by SBP, whereas DBP remained noncontributory. CONCLUSION: Although inferior to the predictive power of SBP, PP was an important determinant of CHD risk in a population of a diverse age span and a better predictor than DBP in men. PMID- 12625601 TI - Antonio Maria Valsalva. PMID- 12625603 TI - Electrocardiographic criteria still useful for mapping of idiopathic right ventricular tachycardia. PMID- 12625602 TI - Electrocardiographic patterns of superior right ventricular outflow tract tachycardias: distinguishing septal and free-wall sites of origin. AB - INTRODUCTION: The superior right ventricular outflow tract (RVOT) septum and free wall are common locations of origin for outflow tract ventricular tachycardias (VT). We hypothesized that (1) unique ECG morphologies of pace maps from septal and free-wall sites in the superior RVOT could be identified using magnetic electroanatomic mapping for accurate anatomical localization; and (2) this ECG information could help facilitate pace mapping and accurate VT localization. METHODS AND RESULTS: In 14 patients with structurally normal hearts who were undergoing ablation for outflow tract VT, a detailed magnetic electroanatomic map of RVOT was constructed in sinus rhythm, then pace mapping was performed from anterior, mid, and posterior sites along the septum and free wall of the superior RVOT. Pace maps were analyzed for ECG morphologies in limb leads and transition patterns in precordial leads. Monophasic R waves in inferior leads for septal sites were taller (1.7 +/- 0.4 mV vs 1.1 +/- 0.3 mV; P < 0.01) and narrower (158 +/- 21 msec vs 168 +/- 15 msec; P < 0.01) compared with free-wall sites; lacked "notching" (28.6% vs 95.2%; P < 0.05); and showed early precordial transition (by lead V4; 78.6% vs 4.8%; P < 0.05). A positive R wave in lead I also distinguished posterior from anterior septal and free-wall sites. Based on QRS morphology in limb leads and precordial transition pattern (early vs late), in a retrospective analysis, a blinded reviewer was able to accurately localize the site of origin of clinical arrhythmia (the successful ablation site on the magnetic electroanatomic map) in 25 of 28 patients (90%) with superior RVOT VT. CONCLUSION: Pace maps in the superior RVOT region manifest site-dependent ECG morphologies that can help in differentiating free-wall from septal locations and posterior from anterior locations. Despite overlap in QRS amplitude and duration, in the majority of patients a combination of ECG features can serve as a useful template in predicting accurately the site of origin of clinical arrhythmias arising from this region. PMID- 12625604 TI - Bigeminy pacing: a new protocol to unmask atrial vulnerability. AB - INTRODUCTION: One of the most exciting developments in our understanding of atrial fibrillation (AF) mechanisms has been the recognition that "AF begets AF" in a process termed atrial remodeling. Little information is available about the events that mediate short-term remodeling. In a bigeminy atrial pacing protocol that produces a continuous extrasystole-postextrasystole cycle length, we sought to evaluate the electrophysiologic consequences of irregular atrial pacing. METHODS AND RESULTS: This study included 22 consecutive patients with documented paroxysmal AF and 10 control subjects. After evaluating the effective refractory period (ERP) and functional refractory period (FRP), bigeminy atrial pacing was performed for 5 minutes. The S1-S2 coupling interval during bigeminy pacing was programmed to a mean value of 275 +/- 45 msec, i.e., 45 msec longer than the basic ERP measured at 100 beats/min. During bigeminy pacing, AF that lasted longer than 1 minute occurred in 12 AF patients and in none of the control subjects (group I). Short salvos of AF occurred in 5 patients and 3 controls (group II). No arrhythmia occurred in 5 patients and 7 controls (group III). Sensitivity, specificity, and negative and positive predictive values of sustained AF induced by bigeminy pacing were 54%, 100%, 50%, and 100%, respectively. No differences were observed between different pacing rates during bigeminy, the premature coupling interval S1-S2, or the conduction parameters S2 A2 and A2. Group I had the shortest basic ERP (222 +/- 38 msec) and group III the longest ERP (242 +/- 21 msec, P < 0.05); group II was intermediate. Atrial ERPs and FRPs measured immediately after termination of 5 minutes of bigeminy pacing were shorter than during baseline. The degree of shortening was similar in AF patients and in controls. The locoregional conduction delay A2 did not change after the bigeminy protocol. CONCLUSION: This study demonstrates that atrial bigeminy pacing highly increases atrial vulnerability. This protocol appears interesting because its sensitivity and specificity are higher than those of the conventional extrastimulation test. This makes it attractive for routine diagnosis of undocumented paroxysmal AF. Because it may induce atrial arrhythmias independently of the classic mechanisms of wavelength shortening, this study emphasizes the need for new modalities in the prevention of atrial arrhythmias. PMID- 12625605 TI - Body surface Laplacian electrocardiogram of ventricular depolarization in normal human subjects. AB - INTRODUCTION: The body surface Laplacian electrocardiogram (ECG) mapping provides a noninvasive means for spatiotemporal mapping of cardiac electrical events. The aim of the present study was to explore the relationship between the Laplacian ECG and the underlying cardiac activities during ventricular depolarization in healthy human subjects. METHODS AND RESULTS: A 95-channel body surface potential ECG was recorded over the anterolateral chest from 11 healthy male subjects. The surface Laplacian (SL) ECG was estimated from the recorded potentials during QRS complex by means of a novel spline SL estimator, as well as by the conventional 5 point SL estimator for comparison purpose. A simulation study was also conducted using a realistic geometry heart-torso model in an attempt to qualitatively interpret the experimental results. For all subjects, more spatial details were observed in the SL ECG maps compared with the potential ECG maps, with spline SL more robust against noise than the 5-point SL. In total, three positive activities (denoted as P1, P2, P3) and four negative activities (denoted as N1, N2, N3, N4) in the spline SL ECG maps were observed during ventricular depolarization. Initial localized P1 and N1 activities were observed in 11 and 8 subjects, respectively. Then, the initial P1 was divided into three positive activities (P1, P2, P3) in 9 subjects. After the appearance of multiple positive activities, three negative activities (N2, N3, N4) appeared in 11, 8, and 9 subjects, respectively. Similar findings were obtained in the computer simulation study. CONCLUSION: The present study demonstrates that the SL ECG provides more spatial details than the potential ECG, and multiple simultaneously active ventricular activities could be revealed in the SL ECG maps. The results suggest that the SL ECG may provide an alternative for noninvasive mapping of cardiac electrical activity. PMID- 12625606 TI - Increased dispersion of refractoriness in patients with atrial fibrillation in the early postoperative period after coronary artery bypass grafting. AB - INTRODUCTION: Increased atrial effective refractory period (AERP) dispersion is well correlated with vulnerability to atrial fibrillation (AF). However, the preoperative electrophysiologic characteristics of atrial abnormalities that may play an important role in the development of AF postoperatively in patients with coronary artery bypass grafting (CABG) have not been investigated in detail. METHODS AND RESULTS: Fifty-six consecutive patients who underwent CABG were enrolled in this study. Eighteen patients (14 men and 4 women; mean age 57.7 +/- 5.2 years) with AF in the early postoperative period and 38 patients (28 men and 10 women; mean age 56.3 +/- 6.4 years) without AF were compared with regard to preoperative clinical, echocardiographic, angiographic, and electrophysiologic parameters. Preoperative PA interval and AERP dispersion values were higher (P < 0.05) in patients who developed AF in the early postoperative period. PA interval (P < 0.05, odds ratio = 1.64, 95% confidence interval 1.17-2.30), AERP in the high right atrium (AERP(HRA); P < 0.05, odds ratio = 0.94, 95% confidence interval 0.91-0.97), AERP in the right posterolateral atrium (AERP(RPL); P < 0.05, odds ratio = 0.79, 95% confidence interval 0.63-0.98), AERP in the distal coronary sinus (AERP(DCS); P < 0.05, odds ratio = 0.84, 95% confidence interval 0.74-1.02), and AERP dispersion (P < 0.001, odds ratio = 1.29, 95% confidence interval 1.12-1.47) were independently related to post-CABG AF in univariate analysis. Increases in preoperative PA interval and AERP dispersion were found to be associated with a high risk for development of post-CABG AF. CONCLUSION: AERP dispersion is a suitable electrophysiologic indicator for atrial vulnerability. The presence of increased preoperative AERP dispersion and PA interval may indicate patients at high risk for development of AF in the early postoperative period after coronary artery bypass grafting. PMID- 12625607 TI - Effect of coronary sinus electrode on the optimal atrial defibrillation pathway for an atrioventricular defibrillator. AB - INTRODUCTION: Previous studies have demonstrated significant failure in converting atrial fibrillation (AF) using a conventional ventricular pathway. The aim of this study was to assess the benefit of incorporating a coronary sinus (CS) lead into the atrial defibrillation pathway in atrial defibrillation threshold (ADFT) reduction in patients with persistent AF. METHODS AND RESULTS: This study was a prospective, randomized assessment of shock configuration on ADFT in 18 patients undergoing elective internal cardioversion for persistent AF (mean AF duration: 8 +/- 9 months). The lead system included a dual-coil defibrillation lead (Endotak DSP, Guidant) with a distal right ventricular (RV) electrode and a proximal superior vena cava (SVC) electrode, a CS lead (Perimeter, Guidant), and a left pectoral cutaneous electrode (Can). In each patient, dual step-up ADFTs were determined for each of three vectors: (1) RV --> SVC+Can; (2) CS --> SVC+Can; and (3) RV --> CS+SVC+Can (group 1, n = 8) or RV+CS -> SVC+Can (group 2, n = 10), using R wave-synchronized biphasic shocks. Successful defibrillation was achieved in all patients without any ventricular proarrhythmia. ADFT of CS --> SVC+Can (11.8 +/- 5.6 J) was significantly lower than ADFT of RV --> SVC+Can (16.5 +/- 7.8 J, P = 0.021). ADFT of CS --> SVC+Can was similar to RV --> CS+SVC+Can (group 1: 12.0 +/- 6.5 J vs 17.4 +/- 4.8 J, P = 0.16), but it was significantly higher than RV+CS --> SVC+Can (group 2: 9.0 +/- 3.9 J vs 11.6 +/- 5.0 J, P = 0.049). CONCLUSION: Patients with persistent AF of substantial duration can be reliably cardioverted using a conventional implantable cardioverter defibrillator (ICD) lead set; however, the incorporation of a CS lead to the conventional ICD lead configuration significantly lowered ADFT. The optimal shock vector that incorporates a CS lead for atrial defibrillation requires future studies. PMID- 12625609 TI - Behavior of nonselective cation channels and large-conductance Ca2+-activated K+ channels induced by dynamic changes in membrane stretch in cultured smooth muscle cells of human coronary artery. AB - INTRODUCTION: The effects of membrane stretch on ion channels were investigated in cultured smooth muscle cells of human coronary artery. METHODS AND RESULTS: In the cell-attached configuration, membrane stretch with negative pressure induced two types of stretch-activated (SA) ion channels: a nonselective cation channel and a large-conductance Ca2+-activated K+ (BK(Ca)) channel. The single-channel conductances of SA cation and BK(Ca) channels were 26 and 203 pS, respectively. To elucidate the mechanism of activation of these SA channels and to minimize mechanical disruption, a sinusoidal change in pipette pressure was applied to the on-cell membrane patch. During dynamic changes in pipette pressure, increases in SA cation channel activity was found to coincide with increases in BK(Ca) channel activity. In the continued presence of cyclic stretch, the activity of SA cation channels gradually diminished. However, after termination of cyclic stretch, BK(Ca) channel activity was greatly enhanced, but the activity of SA cation channels disappeared. CONCLUSION: This study is the first to demonstrate that the behavior of SA cation and BK(Ca) channels in coronary smooth muscle cells is differentially susceptible to dynamic changes in membrane tension. PMID- 12625608 TI - Transvenous defibrillation leads: high incidence of failure during long-term follow-up. AB - INTRODUCTION: Patients with implantable cardioverter defibrillators (ICD) critically depend on correct functioning of their system. The aim of this study was to determine the incidence and clinical presentation of transvenous ICD lead failures during long-term follow-up. METHODS AND RESULTS: The study group consisted of 261 consecutive patients who received Medtronic right ventricular polyurethane transvenous leads (models 6884, 6966, 6936) between 1990 and 1998 as part of an abdominal (n = 70) or pectoral (n = 191) ICD system. During mean follow-up of 4.0 +/- 2.6 years, 31 patients (12%) developed a lead-related sensing failure with oversensing of artifacts. All failures except two were compatible with an insulation defect and occurred late after ICD placement (6.0 +/- 1.8 years after implant). Lead survival decreased from 98% at 4-year follow up to only 62% at 8-year follow-up. Lead survival was not related to patient age, sex, venous lead implantation route, or device implantation site. In 26 (87%) of 31 patients, a sensing defect resulted in inappropriate detection of ventricular fibrillation and subsequent delivery of 3 +/- 3 (range 1-11) inappropriate shocks in 19 (61%) of 31 patients. Device interrogation showed artifacts classified as nonsustained ventricular tachycardia in 21 patients, 40 +/- 43 days before clinically relevant failure of the system. One patient with a subclavian crush syndrome required resuscitation because of undersensing of true ventricular fibrillation. CONCLUSION: Transvenous polyurethane ICD leads showed a high rate of lead insulation failure late after implantation with frequent inappropriate shock deliveries. Close follow-up is mandatory in patients with these leads. Automated device control features with patient alert function integrated into new devices may contribute to early detection of lead failure. PMID- 12625610 TI - Estimation of entrainment response using electrograms from remote sites: validation in animal and computer models of reentrant tachycardia. AB - INTRODUCTION: Studies suggest that entrainment response (ER) of reentrant tachycardia to overdrive pacing can be estimated using signals from sites other than the paced site. METHODS AND RESULTS: A formula for estimation of ER using remote sites against the difference between the postpacing interval (PPI) and tachycardia cycle length (TCL) determined solely from the paced site signal was validated in experimental data and using a simple two-dimensional cellular automata model of reentry. The model also was used to study the behavior and features of entrained surfaces, including the resetting of tachycardia phase by single premature paced stimuli. Experimental results from 1,484 remote sites in 115 pacing sequences showed the average of the median ER estimate error at each pacing site was -2 +/- 5 msec, and the median ER estimate was within 10 msec of PPI-TCL for 94% of pacing sites. From simulation results, ER at the paced site was accurately estimated from >99.8% of 20,764 remote sites during pacing at 24 sites and three paced cycle lengths. Intervals measured from remote electrograms revealed whether the site was activated orthodromically or nonorthodromically during pacing, and results of simulations illustrated that the portion of the surface activated nonorthodromically during pacing increased with distance from the pacing site to the circuit. The phenomenon of nonorthodromic activation of reentrant circuits predicted by modeling was discernible in measurements taken from the animal model of reentrant tachycardia. Results also showed that, for single premature stimuli that penetrated the tachycardia circuit, phase reset of the tachycardia was linearly related to distance between the central obstacle and the paced site. CONCLUSION: The ER is a complex but predictable perturbation of the global activation sequence of reentrant tachycardias. This predictability allows calculations of the response from anywhere on the perturbed surface. These findings suggest new techniques for measurement of the ER, which may lend themselves to computer-based methods for accurate and rapid mapping of reentrant circuits. PMID- 12625611 TI - Entrainment in the age of three-dimensional computer mapping. PMID- 12625612 TI - Three-dimensional mapping of earliest activation after near-threshold ventricular defibrillation shocks. AB - INTRODUCTION: Following shocks with a 50% defibrillation success (DFT50) delivered from electrodes at the right ventricular (RV) apex and superior vena cava (SVC), the earliest epicardial postshock activation always appears focally in the left ventricular (LV) apex for both successful and failed shocks. Because the heart is a three-dimensional (3D) structure, questions remain whether this activation truly arises from a focus or the focal pattern represents epicardial breakthrough resulting from intramural reentry. To answer these questions, 3D electrical mapping was performed. METHODS AND RESULTS: In six pigs, 60 to 84 epoxy fiberglass needles (0.7-mm-diameter), each containing six electrodes 2 mm apart, were inserted into the LV with 3- to 5-mm spacing around the apex and 5- to 10-mm spacing near the base. Ten DFT50 shocks (RV-->SVC, biphasic, 6/4 msec) were delivered after 10 seconds of fibrillation in each animal. The first five activations after each shock were mapped. Of 60 DFT50 shocks, 31 were successful, of which the first postshock cycle was a sinus beat in 13. In the other 18 successful shock episodes, the first postshock activation was detected 63 +/- 16 msec after the shock, which was not significantly different from the 58 +/- 23 msec postshock interval for the 29 failed shock episodes. In these 47 successful and failed shock episodes, the earliest postshock activation always arose focally from the LV apex. Its origin was in the subepicardium in 76% +/- 17%, midmyocardium in 16% +/- 12%, and subendocardium in 8% +/- 6% of cases. CONCLUSION: Following near-DFT50 shocks, the first postshock cycles did not arise by macroreentry. Instead, they originated from a true focus or microreentry, most commonly near the epicardium. PMID- 12625613 TI - Can we learn about ventricular fibrillation in man by studying animal models of defibrillation? PMID- 12625614 TI - Delayed afterdepolarization inhibitor: a potential pharmacologic intervention to improve defibrillation efficacy. AB - INTRODUCTION: Electrical and optical mapping studies of defibrillation have demonstrated that following shocks of strength near the defibrillation threshold (DFT), the first several postshock cycles always arise focally. No immediate postshock reentry was observed. Delayed afterdepolarizations (DADs) have been suggested as a possible cause of this rapid repetitive postshock activity. The aim of this study was to test the hypothesis that DFT is decreased by application of a DAD inhibitor. METHODS AND RESULTS: Six pigs (30-35 kg) were studied. First, control DFT was determined using a three-reversal up/down protocol. Each shock (RV-SVC, biphasic, 6/4 msec) was delivered after 10 seconds of ventricular fibrillation (VF). Then, flunarizine (a DAD inhibitor) was injected intravenously (2 mg/kg bolus and 4 mg/kg/hour maintenance) and the DFT was again determined. A third DFT was determined 50 minutes after drug infusion was terminated to allow the drug to wash out. DFT after flunarizine application (520 +/- 90 V, 14 +/- 3 J) was significantly lower than control DFT (663 +/- 133 V, 23 +/- 4 J). After the drug washed out, DFT (653 +/- 107 V, 22 +/- 4 J) returned to the control DFT value (P = 0.6). Flunarizine reduced the DFT approximately 22% by leading-edge voltage and approximately 40% by energy. CONCLUSION: Flunarizine, a DAD inhibitor, significantly improved defibrillation efficacy. This finding suggests that DADs could be the source of the rapid repetitive focal activation cycles arising after failed near-DFT shocks before degeneration back into VF. Future studies are needed to investigate the cause of the earliest postshock activation and to determine if the DADs are responsible. PMID- 12625615 TI - Emission ratiometry for simultaneous calcium and action potential measurements with coloaded dyes in rabbit hearts: reduction of motion and drift. AB - INTRODUCTION: Optical measurements of the cardiac calcium transient (Ca) and transmembrane action potential (AP) may be performed simultaneously with emission ratiometry to lessen motion artifacts and photobleaching effects. We examined changes in emission spectrum in perfused rabbit hearts coloaded with Rh237 and a green-emitting Ca dye (Fluo-4 or Oregon Green BAPTA 1) to determine wavelength bands for emission ratiometry and to test whether ratiometry reduces motion artifacts and drift. METHODS AND RESULTS: A 488-nm laser illuminated hearts while a spectrofluorometer collected fluorescence from 489 to 838 nm at 1 kHz. Ratiometry with the Ca- and AP-insensitive emission band 663 to 685 nm (IS) as denominator and the Ca-sensitive band 510 to 532 nm as numerator lessened motion artifacts, which was quantified as a 1.4-fold increase in relative amplitude of the Ca (P < 0.05). Ratiometry with the AP-sensitive band 772 to 794 nm as denominator and the IS as numerator produced a 1.7-fold increase in relative amplitude of the AP (P < 0.05). The ratiometry decreased photobleaching-dependent drift by a factor of 0.6 (P < 0.05) for Ca and 0.45 (P < 0.05) for AP. CONCLUSION: Simultaneous Ca and AP emission ratiometry reduces motion artifacts and drift in hearts with coloaded dyes. PMID- 12625616 TI - Automated external defibrillator arrhythmia detection in a model of cardiac arrest due to commotio cordis. AB - INTRODUCTION: Cardiac arrest due to chest wall blows (commotio cordis) has been reported with increasing frequency in children, and only about 15% of victims survive. Automated external defibrillators (AEDs) have been shown to be life saving in adults with cardiac arrest, but data on their use in children are limited. In a swine model of commotio cordis designed to be most relevant to young children, we assessed the efficacy of a commercially available AED for recognition and termination of ventricular fibrillation. METHODS AND RESULTS: Ventricular fibrillation was produced in anesthetized juvenile swine by precordial impact from a baseball under controlled conditions. Animals were randomized to defibrillation after 1, 2, 4, or 6 minutes of ventricular fibrillation. Twenty-six swine underwent 50 ventricular fibrillation inductions. Sensitivity of the AED for recognition of ventricular fibrillation was 98%, and specificity for nonshockable episodes was 100%. All episodes of ventricular fibrillation were successfully terminated by the AED. CONCLUSION: In this experimental model of commotio cordis, the AED proved to be highly sensitive and specific for recognition of ventricular fibrillation and effective in terminating the arrhythmia and restoring sinus rhythm. These findings suggest that early defibrillation with the AED could save young lives on the athletic field. PMID- 12625617 TI - Biventricular implantable cardioverter defibrillator and inappropriate shocks. AB - A 53-year-old man with nonischemic cardiomyopathy underwent implantation of a biventricular implantable cardioverter defibrillator (ICD) for symptomatic ventricular tachycardia. He received five shocks while attempting to exercise, 48 hours after implantation. Interrogation of the device revealed double counting of ventricular sensed events by the left and right ventricular leads. Shortening the AV delay and AV nodal blockade (beta-blocker) to promote ventricular pacing failed to prevent additional inappropriate ICD discharges. After detailed consideration of all options including AV nodal ablation, we chose to disconnect the left ventricular lead pending availability of newer devices with sensing functions limited to the right ventricular lead. Since then, the patient has not experienced any additional inappropriate discharges. PMID- 12625618 TI - Paroxysmal supraventricular tachycardia with persistent ventriculoatrial block. AB - We report the case of a 64-year-old patient with paroxysmal supraventricular tachycardia and persistent VA block. Induction and maintenance of tachycardia occurred without apparent activation of the atria. Diagnostic characteristics were most compatible with AV nodal reentrant tachycardia (AVNRT). Automatic junctional tachycardia and orthodromic nodoventricular or nodofascicular reentry tachycardia were considered in the differential diagnosis. Upper common pathway block during AVNRT may be explained by either intra-atrial conduction block or purely intranodal confined AVNRT. The arrhythmia was cured by a typical posteroseptal ablation approach guided by slow pathway potentials. PMID- 12625619 TI - ATP-sensitive K+ channels of vascular smooth muscle cells. AB - ATP-sensitive potassium channels (K(ATP)) of vascular smooth muscle cells represent potential therapeutic targets for control of abnormal vascular contractility. The biophysical properties, regulation and pharmacology of these channels have received intense scrutiny during the past twenty years, however, the molecular basis of vascular K(ATP) channels remains ill-defined. This review summarizes the recent advancements made in our understanding of the molecular composition of vascular K(ATP) channels with a focus on the evidence that hetero octameric complexes of Kir6.1 and SUR2B subunits constitute the vascular K(ATP) subtype responsible for control of arterial diameter by vasoactive agonists. PMID- 12625620 TI - Sophisticated architecture is required for the sinoatrial node to perform its normal pacemaker function. AB - The heart's pacemaker, the sinoatrial node, does not consist of a group of uniform sinoatrial node cells embedded in atrial muscle. Instead, it is a heterogeneous tissue with multiple cell types and a complex structure. Evidence suggests that from the periphery to the center of the sinoatrial node, there is a gradient in action potential shape, pacemaking, ionic current densities, connexin expression, Ca2+ handling, myofilament density, and cell size. This complexity may be necessary for the sinoatrial node to pacemake under diverse conditions, drive the more hyperpolarized atrial muscle, and resist proarrhythmic perturbations. PMID- 12625621 TI - Incomplete right bundle branch block during supraventricular tachycardia: fact or fiction? PMID- 12625622 TI - Wide QRS complex tachycardia with negative precordial concordance: always a ventricular origin? PMID- 12625623 TI - Pulmonary vein ablation using the LocaLisa nonfluoroscopic mapping system. PMID- 12625624 TI - Cellular mechanisms underlying the long QT syndrome. PMID- 12625625 TI - Patch-clamp experiments with porins extracted from a marine bacterium (Photobacterium profundum strain SS9) and reconstituted in liposomes. AB - The reconstitution of bacterial porins in liposome bilayers for patch-clamp recording is well established. However, the solutions used in the dehydration, rehydration, and osmotic swelling of the liposomes have been developed for porins from enteric bacteria. Porins from marine bacteria normally function in contact with seawater whose ionic composition and osmotic pressure would appear to be incompatible with the established methods. Here, we show that, contrary to expectation, an established reconstitution and patch-clamp method works well with porins, mainly OmpH and OmpL, extracted from the deep-sea marine bacterium Photobacterium profundum strain SS9 and that seawater can be introduced at a supplementary stage. PMID- 12625626 TI - Scorpion toxins from Buthus martensii Karsch all possess a predicted alpha-tight turn. AB - We have purified a new toxin (BmK 17[4]) from Asian scorpion (Buthus martensii Karsch) venom that possesses a distinctive structural motif in its N-terminal (positions 8-12) that is similarly found in two other previously described alpha like toxins. BmK 17[4] prolongs action potentials (APs) in frog nerve and was purified using gel filtration, ion exchange, fast protein liquid chromatography (FPLC), and high-performance liquid chromatography (HPLC). BmK 17[4] significantly prolonged frog APs but it did not alter APs from an insect ventral nerve cord at similar doses. When applied to voltage-clamped frog muscle single fibers, BmK 17[4] prolonged fast inactivation. Because the polypeptide prolongs APs when both K+ and Ca2+ channels were blocked, BMK 17[4] acts to selectively alter Na+ channel inactivation. The N-terminal sequence of BmK 17[4] was found to be VRDAYIAKPENCVYXC --. The molar mass of BmK 17[4] was determined by LC/MS/MS to be 7097 Daltons. The N- terminal motif (KPENC), which introduces a reverse turn in residues 8-12, does not appear in previously characterized BmK alpha-toxins and may be characteristic of alpha-like toxins. Sequence similarity database searches were used to test whether the N-terminal sequences of alpha-like polypeptide toxins from B. martensii Karsch possess a distinctive structural motif in its 5-residue reverse turn (alpha-turn) that is conserved. Sequence similarities with putative polypeptides encoded by cDNAs obtained from a cDNA library [Zhu, S. Y., Li, W. X., Zenq, X. C., et al. (2000) Nine novel precursors of Buthus martensii scorpiox alpha-toxin homologues. Toxicon 38, 1653-1661] from BmK venom glands showed that an active polypeptide toxin cleaved from the putative propolypeptide toxin BmK M9 is likely identical to BmK 17[4]. Sequence comparisons with toxins and putative toxins from B. martensii Karsch and other species revealed that a group of these toxins possess a common structural motif in their alpha-turn. A neighbor-joining phylogenetic analysis suggests that there are two phylogenetic sister groups of related BmK polypeptides; one possesses the KPENC motif and the other possesses a modifed version (KPHNC) of it. PMID- 12625627 TI - Protein dynamics from solution NMR: theory and applications. AB - Solution nuclear magnetic resonance (NMR) spectroscopy is unique in its ability to elucidate the details of atomic-level structural and dynamical properties of biological macromolecules under native-like conditions. Recent advances in NMR techniques and protein sample preparation now allow comprehensive investigation of protein dynamics over timescales ranging 14 orders of magnitude at nearly every atomic site. Thus, solution NMR is poised to reveal aspects of the physico chemical properties that govern the ensemble distribution of protein conformers and the dynamics of their interconversion. We review these advances as well as their recent application to the study of proteins. PMID- 12625628 TI - Cellular and molecular mechanisms of cerebellar granule cell migration. AB - The real-time observation of cell movement in brain slice preparations reveals that in the developing brain, postmitotic neurons alter their shape concomitantly with changes in the mode, direction, tempo, and rate of migration as they traverse different cortical layers. Although it has been hypothesized that orchestrated activities of multiple external cues and cell-cell contact are essential for controlling the cortical-layer-specific changes in cell migration, signaling mechanisms and external guidance cues related to the alteration of neuronal cell migration remain to be determined. In this article, we will first review recent studies on position-specific changes in granule cell behavior through different migratory terrains of the developing cerebellar cortex. We will then present possible roles for the coordinated activity of Ca2+ channels, NMDA type of glutamate receptors, and intracellular Ca2+ fluctuations in controlling cerebellar granule cell movement. Furthermore, we will discuss the crucial roles of brain-derived neurotrophic factor (BDNF), neuregulin (NRG), stromal cell derived factor 1alpha (SDF-1alpha), ephrin-B2, and EphB2 receptor in providing directional cues promoting granule cell migration from the external granular layer (EGL) to the internal granular layer (IGL). Finally, we will demonstrate that endogenous somatostatin controls the migration of granule cells in a cortical layer-specific manner: Endogenous somatostatin accelerates granule cell movement near the birthplace within the EGL, but significantly slows down the movement near their final destination within the IGL. PMID- 12625629 TI - Mechanotransduction determines the structure and function of lung and bone: a theoretical model for the pathophysiology of chronic disease. AB - Multicellular organisms have evolved in adaptation to the Earth's gravitational and oxygen environment. This epigenetic process is dependent on the capacity of mesodermal cells to act as mechanosensors that can conform, deform, and reform in adaptation to the organism's physical environment. Mechanical forces, such as hydrostatic pressure and gravity, play important roles in the embryonic development, homeostasis, and repair of lung and bone. We discuss the role of parathyroid hormone-related protein (PTHrP) as a mechanotransducer for stretch in these organs during normal development, particularly as it lends itself to homeostasis; we further demonstrate that "uncoupling" of such mechanisms may play a central role in injury repair, particularly as it relates to chronic diseases of lung and bone. Endothermal PTHrP signaling through its G-protein coupled receptor promotes normal cell-cell signaling that maintains the homeostatic phenotypes of lung and bone. Molecular disruption of the PTHrP/PTHrP receptor pathway from endoderm to mesoderm, because of such factors as volutrauma, hyperoxia, inflammation, and microgravity, alters intracellular signaling, causing maladaptive cellular changes, resulting in myofibroblast proliferation and granulation. Examples of such pathologic changes specifically related to this cellular/molecular mechanism of maladaptation are chronic lung disease and osteoporosis. We suggest a new paradigm that may help in the future creation of diagnostic and therapeutic modalities for a wide range of developmental and chronic diseases ranging from bronchopulmonary dysplasia in newborns to idiopathic pulmonary fibrosis and osteoporosis as a result of aging or microgravity. PMID- 12625630 TI - How we got to the Women's Health Initiative hormone replacement trial. PMID- 12625631 TI - The Women's Health Initiative estrogen plus progestin trial: the study and how it changes our practice. PMID- 12625632 TI - Focus on cardiovascular health: hormone replacement therapy, estrogen replacement therapy, and selective estrogen receptor modulators. PMID- 12625633 TI - Prevention and treatment of osteoporosis: where are we today? PMID- 12625634 TI - Motor recovery after stroke: new directions in scientific inquiry. AB - The Decade of the Brain is now over, but the most important discoveries are still to come. Given that in the United States and elsewhere in the world the population is aging, the impact of high-quality brain research over the next 2 decades could be tremendous. It is important to improve stroke prevention and acute stroke treatment and also to find the means to eliminate stroke-related disability for those who live each day with motor and other neurologic impairments. PMID- 12625635 TI - Overview of cortical plasticity and recovery from brain injury. AB - The analysis of plastic changes in the nervous system is based on the assumption that the nervous system is not a static system but rather one that changes over time. Neural plasticity can be studied at many levels, beginning with behavior and then becoming progressively more microscopic by descending to the level of cerebral maps, synaptic organization, physiologic activities, molecular structure, and mitosis. This article considers each level in turn and then briefly describes how an understanding of the principles of plasticity can be used to initiate treatments for cerebral injury. PMID- 12625636 TI - Should the injured and intact hemispheres be treated differently during the early phases of physical restorative therapy in experimental stroke or parkinsonism? AB - Over a century ago the intact cortex was proposed to contribute to recovery from unilateral brain injury, but its possible role in functional outcome has become more appreciated in recent years as a result of anatomic, metabolic and behavioral studies. Although use of the contralesional limb is naturally impaired after sensorimotor cortex injury, neural and astrocytic events in the intact hemisphere may give rise to, and may be influenced by, an enhanced ability to compensate for lost motor function. The debate is still open as to whether the neural changes are generally compensatory in nature, with activity in the homotopic cortex leading to greater capability in the nonimpaired limb, or whether they are actually a matter of reorganization in the homotopic cortex leading to connections to denervated targets in the opposite hemisphere, thus allowing the homotopic cortex to control motor programs there. Although both phenomena may occur to some degree, there is mounting evidence in support of the former view. Careful behavioral techniques have been developed that can expose compensatory tricks, and the time course of these behaviors correlates well with anatomic data. Moreover, if the intact cortex sustains a second lesion after recovery from the first, forelimb sensorimotor function specific to the first impaired side of the body is not worsened. Partial denervation of callosal fibers coming from the injured hemisphere, plus preferential use of the good forelimb caused by a cortical injury, may increase trophic factors in the intact hemisphere. These and related events seem to provide a growth-favorable environment there that permits motor learning in the intact forelimb at a level of skill exceeding that which a normal animal can attain in the same period of time. There are anecdotal cases in human neurologic patients that are consistent with these findings. For example, a colleague of the authors who sustained a unilateral infarction that rendered his dominant right hand severely impaired noticed that soon after the stroke he was able to use his left hand for writing and computers as well as he had ever used his right hand. Cross-midline placing tests also indicate that the structural events observed in the intact cortex may potentiate projections to the damaged hemisphere. These changes may help restore the capacity of tactile information projecting to the intact hemisphere to control limb placing in the impaired forelimb. Neural events in the injured hemisphere can be affected by behavior differently than the neural events in the intact hemisphere. Different therapeutic strategies might well be used on opposing limbs at different times after unilateral sensorimotor cortex injury to optimize recovery (and, indeed, to avoid exaggerating the insult). Finally, the details of reorganization in both hemispheres differ greatly depending on the type of brain injury sustained (eg, in stroke versus Parkinson's disease), suggesting that an approach that considers the role of both hemispheres is likely to be beneficial in research on a broad variety of brain pathologies. PMID- 12625637 TI - Functional magnetic resonance imaging in stroke recovery. AB - A better understanding of brain reorganization following stroke may have value in predicting outcome defining the targets of restorative therapies, measuring the physiology of recovery, and in serving as a biologic marker in studies targeting stroke recovery. Measures of brain organization may provide insights as to which patients retain relevant substrate for therapies that target restorative events. Brain reorganization can be studied with many different methods that often have complementary value. Functional MR imaging provides insights into brain plasticity after stroke and remains a valuable tool in the study of motor recovery after stroke. PMID- 12625638 TI - Functional and structural plasticity in motor cortex: implications for stroke recovery. AB - Several studies have now demonstrated that the motor cortical representations are dynamically maintained in both normal and brain-injured animals. Functional plasticity in the motor cortex of normal animals is accompanied by changes in synaptic morphology; these changes are skill-dependent rather than simply use dependent. Finally, motor cortical areas undergo substantial functional alterations after focal ischemic infarcts; motor experience is a potent and adaptive modulator of injury-related plasticity. These recent neuroscientific advances set the stage for the development of new, more effective interventions in chronic stroke populations that are based on the basic mechanisms underlying neuroplasticity. PMID- 12625639 TI - Improved motor recovery after stroke and massive cortical reorganization following Constraint-Induced Movement therapy. AB - Constraint-Induced Movement therapy (CI therapy) has been demonstrated to improve motor function and upper extremity (UE) use of persons with hemiparesis resulting from chronic stroke through two separate but linked mechanisms, overcoming learned nonuse, and facilitating use-dependent cortical reorganization. The principles of CI therapy and adaptations of the basic techniques have been used successfully with diagnostic categories other than stroke that involve disability greater than what is warranted by the organic condition of the individual. Because neuroimaging and transcranial magnetic stimulation studies indicate that many of these conditions involve abnormalities of cortical organization, CI therapy might therefore be viewed as a technique that achieves clinical efficacy by correcting disorders of brain plasticity. CI therapy constitutes a new approach to neurorehabilitation and, with continued investigation, elaboration, and application to clinical settings, it seems to hold considerable promise. PMID- 12625640 TI - Neuromuscular electrical stimulation for motor relearning in hemiparesis. AB - Neuromuscular electrical stimulation may have an important role in improving the motor function of stroke survivors. Active, repetitive movement training mediated by transcutaneous cyclic and EMG-triggered NMES may facilitate the motor recovery of stroke survivors. Multicenter, double-blinded, randomized clinical trials should be pursued to confirm the motor-relearning effects of transcutaneous NMES and to define appropriate prescriptive specifications. Intramuscular EMG controlled NMES may be superior to transcutaneous systems and is presently undergoing preliminary randomized clinical trials. Neuroprostheses systems may provided the highest level of goal-oriented activity and cognitive investments, which may lead to significant motor relearning. Implementation of clinically viable neuroprosthesis systems, however, will probably require additional technical developments including more reliable control paradigms and methods for blocking undesirable muscle contractions. In view of the dynamic nature of the present health care environment, the future of NMES technology is difficult to predict. By necessity, scientists and clinicians must continue to explore new ideas and to improve on the present systems. Components will be smaller, more durable, and more reliable. Control issues will remain critical for both motor relearning and neuroprosthetic applications, and the implementation of cortical control is likely to dictate the nature of future generations of NMES systems. Finally, consumers will direct future developments. In the present health care environment, where cost has become an overwhelming factor in the development and implementation of new technology, the consumer will become one of technology's greatest advocates. The usual drive toward greater complexity will be tempered by the practical issues of clinical implementation, where patient acceptance is often a function of a tenuous balance between the burden or cost associated with using a system and the system's impact on the user's life. PMID- 12625641 TI - Treadmill training with partial body weight support after stroke. AB - Treadmill therapy with partial BWS is a promising new approach to improve gait ability after stroke. This task-specific approach enables nonambulatory patients the repetitive practice of complex gait cycles instead of single-limb gait preparatory maneuvers. Patients walk more symmetrically with less spasticity and better cardiovascular efficiency on the treadmill than with floor walking. Several controlled, clinical studies have shown the potential of treadmill training as a therapeutic intervention for nonambulatory patients with chronic stroke-related hemiplegia. Furthermore, controlled trials in acute stroke survivors have shown that treadmill training is as effective as other physiotherapy approaches that stress the repetitive practice of gait. Controlled multicenter trials comparing locomotor training with conventional therapy will be forthcoming. An electromechanical gait trainer that relieves the strenuous effort of the therapists and provides control of the trunk in a phase-dependent manner is a new technical alternative for gait training in severely impaired stroke patients. PMID- 12625642 TI - Amphetamines and related drugs in motor recovery after stroke. AB - Studies in laboratory animals indicate that the rate and extent of functional recovery after focal brain injury can be modulated by drugs affecting specific central neurotransmitters. Preliminary clinical studies suggest that similar drug effects may occur in humans recovering from stroke. Combined with principles derived from the laboratory, these clinical studies provide important insights to guide the rational design of trials aimed at determining the clinical use of this approach to improving poststroke recovery. PMID- 12625643 TI - Growth factors and stem cells as treatments for stroke recovery. AB - Both polypeptide growth factors and stem cell populations from bone marrow and umbilical cord blood hold promise as treatments to enhance neurologic recovery after stroke. Growth factors may exert their effects through stimulation of neural sprouting and enhancement of endogenous progenitor cell proliferation, migration, and differentiation in brain. Exogenous stem cells may exert their effects by acting as miniature "factories" for trophic substances in the poststroke brain. The combination of growth factors and stem cells may be more effective than either treatment alone. Stroke recovery represents a new and relatively untested target for stroke therapeutics. Whereas acute stroke treatments focus on agents that dissolve blot clots (thrombolytics) and antagonize cell death (neuroprotective agents), stroke recovery treatments are likely to enhance structural and functional reorganization (plasticity) of the damaged brain. Successful clinical trials of stroke recovery-promoting agents are likely to be quite different from trials testing acute stroke therapies. In particular, the time window of effective treatment to enhance stroke recovery is likely to be far longer than that for acute stroke treatments, perhaps days or weeks rather than minutes or hours after stroke. This longer time window means that time is available for careful screening and testing of potential subjects for stroke recovery trials, both in terms of size and location of cerebral infarcts and in type and severity of neurologic deficits. Detailed baseline information can be obtained for each patient against which eventual clinical outcome can be compared. Finally, separate and detailed outcome measures can be obtained in both the sensorimotor and cognitive neurologic spheres, because it is possible that these two kinds of function may recover differently or be differentially responsive to recovery-promoting treatments. Stroke recovery represents an important and underexplored opportunity for the development of new stroke treatments. PMID- 12625644 TI - The therapeutic effects of cellular therapy for functional recovery after brain injury. AB - The studies presented in this article suggest that marrow-derived cellular therapy may be an effective adjunct treatment for functional recovery after stroke. Cellular therapy can enhance the endogenous restorative mechanisms of the injured brain, assisting the tissue as it returns to a "developmental" state and supporting the process of neovascularization, neurogenesis, and neural reorganization. The advantages of using MSCs are that they can be given as an autologous graft, avoiding risks of rejection and graft-versus-host reactions, and that they can be given intravenously, minimizing complications. It is anticipated that cellular therapy, in combination with standard rehabilitation therapy and neural retraining, can improve functional outcomes following stroke. PMID- 12625645 TI - Neuronal transplantation for motor stroke: from the laboratory to the clinic. AB - Laboratory studies have established the potential for neuronal transplantation to be of benefit to patients. Experimental studies in normal animals indicate that brain implantation of neurons seems safe. Implanted neurons integrated with the host brain, sent out axonal processes to communicate with other nerve cells, released transmitters (the chemical messengers of nerve cell communication), and demonstrated typical neuronal proteins. This article discusses phase I and II trials of neuronal transplantation in humans with small strokes in critical brain locations such as the basal ganglia region. More work is needed to confirm safety and to identify optimal measures of efficacy in this setting. PMID- 12625646 TI - Experiential learning in health administration: a new approach for new times. AB - A balance of experiential and didactic learning is essential to develop managers and leaders for early and mid-career positions in the healthcare administration profession. This article suggests pedagogical methods that enhance experiential learning. It also recognizes the need for a renewed focus on diverse experiential learning opportunities, management and preceptor development programs, and graduate program partnerships. PMID- 12625647 TI - Competency-based health services management education: contemporary issues and emerging challenges. AB - Health service executives increasingly recognize managerial competencies as an important element of contemporary health services management education and practice. This paper addresses reasons for the growing interestin the topic, provides a brief review of selected literature, and presents findings from two Delphi studies of the identification and use of competencies in health services management. The two studies reveal that there is a high level of agreement among academicians and practitioners concerning which domains of competencies are effectively addressed in health management education, but many still have concerns regarding how the competencies should be applied. The paper concludes with unresolved issues and their implications for future research. PMID- 12625648 TI - The dominant influence of social context on CEO learning in health care: a challenge to traditional management continuing education and development. AB - Little is known about their learning preferences of the health care industry's most senior executives, particularly CEOs. This paper focuses on the "powerful mechanisms for CEO learning" made available to health care managers through "exclusive learning groups." By meeting ad hoc or regularly under the auspices of such organizations as The Aspen Institute, the American College of Health Care Executives, Healthcare Research and Development, Inc., and other professional and commercial organizations, CEOs come together to discuss the challenges and dilemmas of their jobs. This paper reports on a two-part research effort to determine why CEO exclusive learning groups are so effective. A survey of 70 CEOs in health care-related organizations and in-depth interviews with 8 CEOs reveals that CEOs use exclusive learning group sessions as social learning vehicles, as opportunities to create secondary-level executive networks, as mechanisms for achieving a competitive advantage, and as safe, confidential environments to try out new ideas, seek information, and build relationships. PMID- 12625649 TI - A conversation with Patrick G. Hays, MHA, FACHE. PMID- 12625650 TI - Selection criteria for rehabilitation services. AB - In an era of inadequate resources for all health services, rehabilitation services providers are frequently asked to produce criteria defining the patients they will accept. In principle rationing of services by ability to benefit is fair. In practice there are many difficulties. The evidence to allow selection of patients for rehabilitation does not exist and probably the best criterion of ability to benefit from rehabilitation (not 'likely to have a good outcome') is demonstration of benefit within a rehabilitation programme. Basic considerations of epidemiological statistical facts also show that criteria are likely to select too many inappropriate patients into services or exclude too many appropriate patients from services. The solution is for purchasers and providers to develop mutual trust and, in the UK at least, to ensure that patients who do not need rehabilitation can be discharged quickly into appropriate support services. PMID- 12625651 TI - Prognostic factors for ambulation and activities of daily living in the subacute phase after stroke. A systematic review of the literature. AB - OBJECTIVE: To identify evidence-based prognostic factors in the subacute phase after stroke for activities of daily living (ADL) and ambulation at six months to one year after stroke. DESIGN: Systematic literature search designed in accordance with the Cochrane Collaboration criteria with the following data sources: (1) MEDLINE, EMBASE, CINAHL, Current Contents, Cochrane Database of Systematic Reviews, Psyclit, and Sociological Abstracts. (2) Reference lists, personal archives, and consultation of experts. (3) Guidelines. METHODS: Inclusion criteria were: (1) cohort studies of patients with an ischaemic or haemorrhagic stroke; (2) inception cohort with assessment of prognostic factors within the first two weeks after stroke; (3) outcome measures for ADL and ambulation; and (4) a follow-up of six months to one year. Internal, statistical and external validity of the studies were assessed using a checklist with 11 methodological criteria in accordance with the recommendations of the Cochrane Collaboration. RESULTS: From 1,027 potentially relevant studies 26 studies involving a total of 7,850 patients met the inclusion criteria. Incontinence for urine is the only prognostic factor identified in three studies with a level A (i.e., a good level of scientific evidence according to the methodological score). The following factors were found in one level A study: initial ADL disability and ambulation, high age, severe paresis or paralysis, impaired swallowing, ideomotor apraxia, ideational apraxia, and visuospatial construction problems; as well as factors relating to complications of an ischaemic stroke, such as extraparenchymal bleeding, cerebral oedema and size of intraparenchymal haemorrhage. CONCLUSIONS: The present evidence concerning possible predictors in the subacute stage of stroke has insufficient quality to make an evidence-based prediction of ADL and ambulation after stroke because only one prognostic factor was demonstrated in at least two level A studies, our cut-off for sufficient scientific evidence. PMID- 12625652 TI - Obstacle training programme for individuals post stroke: feasibility study. AB - OBJECTIVE: The purposes were threefold: to develop an obstacle ambulation training programme, to recruit and retain individuals post chronic stroke and to assess the effectiveness of the obstacle ambulation training programme. DESIGN: Intervention study; case series; baseline to post-intervention measures and one month follow-up after intervention. SETTING: An outpatient department. SUBJECTS: Five individuals with chronic stroke with mild to moderate functional limitations; convenience sample. INTERVENTIONS: Therapy consisted of twice weekly sessions for four weeks during which subjects walked along a walkway and over obstacles. MAIN OUTCOME MEASURES: Motor Assessment Scale--walking section (MAS- time), Six-minute Walk Test distance, walking velocity, and Medical Outcomes Study-36 Health Status Measurement (SF-36). RESULTS: Significant improvements were seen baseline to post intervention for walking velocity, Six-minute Walk Test, MAS and SF-36 physical function score (p < or = 0.025 for all measures). Most improvements noted at the end of training were retained one month later. The effect sizes ranged from 0.33 to 1.20. CONCLUSIONS: Substantial improvement in ambulation function and disability level were seen as a result of the obstacle training programme for individuals with chronic stroke. Further investigation is warranted. PMID- 12625653 TI - Effects of ankle-foot orthoses on hemiparetic gait. AB - OBJECTIVE: Ankle-foot orthoses (AFOs) are widely used to provide optimal ambulation in people with hemiplegia. In this study we evaluated the mechanical effects of metallic and plastic AFOs on severely hemiparetic stroke patients. METHODS: Twelve hemiparetic patients were analysed on a Vicon 370 Motion Analysis System. Spatiotemporal, kinematic and kinetic parameters were measured. RESULTS: The two types of orthoses generally had similar positive effects on hemiplegic gait parameters, increasing cadence, walking speed, single and double step length, ankle dorsiflexion angle at heel strike and swing. The metallic AFO was better at increasing the ankle dorsiflexion angle than the plastic AFO. CONCLUSION: Hemiplegic gait was improved by both orthoses. However, metallic AFOs provided better stabilization of the ankle, allowing improved heel strike and push-off. PMID- 12625654 TI - Target board test for the quantification of ataxia in tremulous patients. AB - OBJECTIVE: The objective was to develop and assess the validity and reliability of a target board test (TBT) for quantifying ataxia and measuring dysmetria in the presence of tremor. DESIGN: Each subject was instructed to reach out and mark a target placed at arm's length with a pen (10 times with each hand). Ten patients performed the test twice. SETTING: A hospital-based multiple sclerosis (MS) unit. SUBJECTS: Fifty-three patients with MS and upper limb tremor/ataxia and 20 healthy control subjects. The MS patients were classified into four subgroups: MS control group (n = 13), MS tremor group (n = 9), MS dysmetria group (n = 6), MS mixed (tremor and dysmetria) group (n = 25). MAIN OUTCOME MEASURES: The main outcome measures were the average radial distance away from the target (mean R) and the mean directional error (mean V) of the 10 contact points from the target. From these a dysmetria tremor index (DTI) was calculated by dividing mean V by mean R. Also used were a dysmetria scale, a dysdiadochokinesia scale and a finger-tapping test. RESULTS: Mean R correlated significantly with dysmetria, dysdiadochokinesia, kinetic tremor and (inversely) with the finger tapping test (all p < 0.005). The median difference between two measurements of mean R for all 10 contact points was 11.3% and 19.0% and for mean V48.3% and 63.4% and DTI 57.2% and 50.5% for the right- and left-hand sides respectively, indicating the considerable directional variability within ataxia. CONCLUSION: The TBT provides simple quantitative objective measurements of upper limb ataxia. PMID- 12625655 TI - Screening for ability to complete a questionnaire: a preliminary evaluation of the AbilityQ and ShoulderQ for assessing shoulder pain in stroke patients. AB - OBJECTIVES: To assess the repeatability of the AbilityQ--a screening tool to assess technical ability to complete a questionnaire--in patients with complex disabilities following stroke. To evaluate the validity of the AbilityQ in predicting repeatability and consistency of response to verbal and visual analogue scale questions in the ShoulderQ in those with shoulder pain. DESIGN: In a cross-sectional study, the AbilityQ and 'ShoulderQ' were administered on two consecutive days by a single investigator. SETTING: Six regional rehabilitation centres in the UK managing patients with severe complex neurological disability. SUBJECTS: Forty-nine patients with anterior or posterior circulation strokes: 13 (63%) had cognitive deficits and 28 (57%) had communicative deficits. RESULTS: Thirty-one (63.3%) patients required some degree of help in completing the AbilityQ. Percentage agreement for individual questions between day 1 and day 2 ranged from 55 to 88% with kappa values from 0.07 to 0.79. Repeatability of ShoulderQ responses was 36-72% for the verbal questions (kappa values 0.16-0.56). For visual analogue scale (VAS) questions, agreement +/- 1 on a 10-point scale was 36-59% with intraclass correlation coefficients 0.50-0.60 (p < 0.01). High verbal (> or = 3/4) and VAS scores (3/3) on the AbilityQ had positive predictive values for good repeatability in the ShoulderQ of 80% and 58% respectively. Mismatched AbilityQ responses to verbal and VAS questions showed a positive predictive value of 92% for similar inconsistency in the ShoulderQ and a negative predictive value of 90%. CONCLUSIONS: In this preliminary evaluation the AbilityQ and the ShoulderQ each demonstrated a moderate level of repeatability in a group of patients expected to have difficulty in completing questionnaires. The AbilityQ appears to have some clinical usefulness in identifying those able to respond to the ShoulderQ and, where responses to verbal and VAS questions conflict, may offer some guidance as to which are most likely to be accurate in that individual. Further development is underway. PMID- 12625656 TI - Natural history of cognitive impairment after stroke and factors associated with its recovery. AB - OBJECTIVES: To describe the natural history of cognitive impairment following stroke over three years; determine factors associated with recovery of post stroke cognitive impairment and examine the effect of this recovery on stroke outcomes. DESIGN: Observational study. SETTING: Population-based register of first-ever strokes. MEASURES: Mini-Mental State Examination (MMSE) for cognition (cognitive impairment: MMSE<24), Barthel and Frenchay activity indices for disability. METHODS: To describe its natural history, cognition was assessed at three months, 1, 2 and 3 years after stroke in 163 subjects registered in 1995. Recovery of cognitive impairment was examined using a larger cohort registered between 1995 and 1998 (n = 476) and 193 subjects cognitively impaired at three months were reassessed a year post stroke; 34 who had recovered cognitively (MMSE 24-30) were compared with 102 with persistent cognitive impairment in terms of demography, risk factors, initial impairments and stroke subtype. RESULTS: At three months, 1, 2 and 3 years post stroke, the prevalence rates of cognitive impairment were 39%, 35%, 30% and 32% respectively. Multivariable analyses showed that recovery was associated with smoking (OR 3.7; 95% CI 1.2-11.8), compromised by visuospatial neglect (OR 0.27; 95% CI 0.08-0.89), and had a near-significant association with right hemispheric lesion (OR 2.87; 95% CI 0.94-8.78). Cognitive recovery was associated with less institutionalization (p = 0.032) and being less disabled, on Barthel (p = 0.001) and Frenchay Activity Indices (p = 0.028). CONCLUSIONS: Cognitive impairment remains highly prevalent up to three years after first stroke. Recovery from cognitive impairment is associated with smoking and possibly right hemisphere strokes, but compromised by visuospatial neglect. These associations require further clarification. PMID- 12625657 TI - Changing patterns of cognitive-motor interference (CMI) over time during recovery from stroke. AB - OBJECTIVE: To explore the extent and nature of change in cognitive-motor interference (CMI) among rehabilitating stroke patients who showed dual-task gait decrement at initial assessment. DESIGN: Experimental, within-subjects, repeated measures design. SETTING: Rehabilitation centre for adults with acquired, nonprogressive brain injury. SUBJECTS: Ten patients with unilateral stroke, available for reassessment 1-9 months following their participation in a study of CMI after brain injury. MEASURES: Median stride duration; mean word generation. METHODS: Two x one-minute walking trials, two x one-minute word generation trials, two x one-minute trials of simultaneous walking and word generation; 10 metre walking time; Barthel ADL Scale score. RESULTS: Seven out of ten patients showed reduction over time in dual-task gait decrement. Three out of ten showed reduction in cognitive decrement. Only one showed concomitant reduction in gait and word generation decrement. CONCLUSION: Extent of CMI during relearning to walk after a stroke reduced over time in the majority of patients. Effects were more evident in improved stride duration than improved cognitive performance. Measures of multiple task performance should be included in assessment for functional recovery. PMID- 12625658 TI - A hypothesis: self-propulsion in a wheelchair early after stroke might not be harmful. AB - BACKGROUND: There is often strong clinical resistance to patients self-propelling a wheelchair post stroke as this is believed to produce immediate increases in abnormal posture and movement. Research to support this viewpoint is limited. OBJECTIVE: To begin investigation of the immediate effects of self-propulsion on symmetrical sitting. DESIGN: Replicated single-case studies ABABA. SETTING: Movement analysis laboratory. SUBJECTS: Four patients, a maximum of eight weeks post stroke and six age-matched healthy volunteers. INTERVENTIONS: Subjects sat in the wheelchair during the A phases and self-propelled forwards during the B phases. The Manchester Active Position Seat (consists of 68 force transducers which transmit data at 10 Hz) measured the magnitude of peak force and the position of peak force on both sides of the seat. The mean symmetry index and standard deviation for each study phase were calculated and graphed for each subject. Interpretation was by visual inspection. RESULTS: Only one stroke patient and one volunteer increased asymmetry of magnitude of peak force following the two periods of self-propulsion. Only one of the stroke patients increased asymmetry of position of peak force following self-propulsion compared with three of the healthy volunteers. CONCLUSIONS: These results raise the hypothesis that self-propulsion early post stroke might not produce immediate detrimental effects on seated symmetry. PMID- 12625659 TI - Use of unconventional therapies by individuals with multiple sclerosis. AB - OBJECTIVE: To examine the prevalence and patterns of use of complementary and alternative medicine (CAM) among individuals with multiple sclerosis (MS) in the USA and to explore the reasons for use, symptoms treated and perceived effectiveness of these therapies. METHODS: Surveys were mailed to the entire mailing list of the MS Foundation, constituting 11,600 individuals with MS or their family members; 3,140 adults with MS returned surveys, yielding a response rate of 27.1%. RESULTS: More than half of the responding sample (57.1%) had used at least one CAM modality. The longer that people had MS and the less satisfied they were with conventional health care the more likely they were to use CAM therapies. The most common reasons for using CAMs were the desire to use holistic health care (i.e., treatments that recognized the interrelatedness of mind, body and spirit) and dissatisfaction with conventional medicine. Ingested herbs were the most frequently used CAM modalities (26.6%), followed by chiropractic manipulation (25.5%), massage (23.3%) and acupuncture (19.9%). Women were 25% more likely than men and whites were 30% more likely than non-whites to use CAM therapies. There was no significant relationship between the frequency of use and the reported efficacy of the CAM techniques (r = 0.17, p > 0.10). CONCLUSIONS: The prevalence of CAM use in this population warrants more research on the efficacy and safety of these therapies, especially those with high usage or high efficacy ratings, such as herbs, chiropractic manipulation and massage, but for which there is little or no research evidence for efficacy or safety. PMID- 12625660 TI - Life goals: the concept and its relevance to rehabilitation. AB - OBJECTIVE: Life goals are desired states that people seek to obtain maintain or avoid. These goals may influence motivation to participate in the rehabilitation process. The aim of this paper is to review the literature on life goals and the influence of life goals on the rehabilitation process. METHODS: The MEDLINE, EMBASE, Psychlit and CINAHL databases were searched with the keywords goals, life goals, aim of life, meaning of life, motivation, assessment (identification) of life goals, goal planning, disability, coping and rehabilitation. RESULTS: The initial search produced 917 abstracts. After going through these abstracts, 39 articles were selected for inclusion in the review. Age, gender, personality, experiences and society and environment influence life goals. Pursuit and attainment of life goals affect sense of well-being. Life goals are accessible to conscious awareness and can be identified. Several questionnaires are available for assessment of life goals. Different questionnaires assess different aspects of life goals. All except one of these questionnaires need to be tested for validity and reliability in a rehabilitation setting. Disabilities interfere with goal striving and result in emotional distress. Motivation to participate in a rehabilitation programme depends on concurrence between a patient's life goals and treatment goals. Incorporation of a subject's life goals into a management programme resulted in better outcomes in various physical and psychiatric disorders. There are no data on the efficacy of life goal-orientated rehabilitation programmes. CONCLUSIONS: Life goals influence patients' motivation to participate in and compliance with treatment programmes. We still do not know whether rehabilitation programmes focusing on life goals make any difference in outcome. There is need for further studies in this area. PMID- 12625661 TI - Efficacy of a specially designed hip protector for hip fracture prevention and compliance with use in elderly Hong Kong Chinese. AB - BACKGROUND: Hip protectors, while effective in prevention of hip fractures, have been designed for a Caucasian body build and may not be suitable for Asian subjects living in a subtropical climate. OBJECTIVE: Hip protectors and accompanying shorts were specially designed for use for the Chinese body build and subtropical climate, and tested for compliance and efficacy in fracture prevention. METHOD: A pragmatic study examining compliance with the wearing of hip protectors, and effectiveness in preventing hip fractures (using a case control design), in 302 subjects wearing hip protectors and 352 control subjects. RESULTS: Overall compliance varied from 55 to 70%. The relative risk for hip fracture was 0.18 (0.04-0.79), relative risk reduction 82%, and the number of subjects needing to wear a hip protector to prevent one fracture was 33 (19-117). CONCLUSION: Hip protectors adapted for use in Chinese people appear to reduce hip fractures in routine clinical practice. PMID- 12625662 TI - Concurrent validity of an electronic descriptive pain scale. AB - The study objective was to assess the concurrent validity of the Electronic Descriptive Pain Scale (EDPS), a pain scale built into a transcutaneous electrical nerve stimulation device. One hundred patients in an outpatient physiotherapy (PT) clinic participated (mean age 41.30 years, SD 13.95). Before and after a PT treatment, subjects rated their current pain intensity with the EDPS, a visual analogue scale, a numerical pain rating scale, and the McGill Pain Questionnaire's Present Pain Intensity. The results showed relatively high significant correlations between the EDPS and each of the other pain scales. PMID- 12625663 TI - Duration of condition is unrelated to health-state valuation on the EuroQoL. AB - OBJECTIVE: To determine whether health valuations, such as those used in economic evaluation, are affected by duration of a health condition. People with disabling health conditions tend to value health more highly than members of the general population, and one explanation for this is that over time their experience of living with a disabling illness changes the way in which they value health. If this is so, a relationship between the duration of an individual's disabling health condition and the valuation they assign to their health-state might reasonably be expected. DESIGN: A postal survey using the EuroQoL (EQ-5D) instrument to collect descriptions and valuations for health from people who reported a diagnosis of either stroke or multiple sclerosis. Contact with participants was made through national support organizations and questionnaires were returned by mail. RESULTS: Eight hundred and ninety-four people completed the survey. One hundred (11 %) had one health-state indicating moderate problems in all five dimensions of the EQ-5D descriptive profile. For people with this health-state, analysis of covariance showed no relationship between valuation of health-state and time from onset of illness (F = 0.38, p = 0.54). This finding applied irrespective of the diagnosis, and for some other less frequently reported health-states. CONCLUSION: Clinical experience suggests that over time people adapt to long-term disability. However we found no evidence to support the proposition that higher health-state valuations by people with disabling conditions are explained by the actual duration of their condition. PMID- 12625664 TI - A regression analysis of the Waterlow score in pressure ulcer risk assessment. AB - OBJECTIVE: To explore the predictive value of the Waterlow score, and the subscores of age and gender. DESIGN: Logistic regression analysis was conducted on the two subscores of the Waterlow score, and the residual Waterlow score with gender and age removed. Receiver operating characteristic (ROC) analysis gave a quantitative measure of the classification ability of the Waterlow score. SETTING: Burton, UK. SUBJECTS: All admissions over a five-year period to the District General Hospital, a total of 150,015 admissions of 82,691 patients. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Area under the ROC curve for significant (as determined by logistic regression) variables. RESULTS: Data were inaccurate in at least 44.7% of the records, and analysis was conducted on the 43,735 records for which no errors were apparent. Nine hundred and fifty-four patients had a pressure ulcer on admission (2.1%); 277 developed a pressure ulcer (0.6%). The Waterlow score was predictive of pressure ulcers. Age was predictive, and gender was not found to be a significant predictor. CONCLUSIONS: The Waterlow score may be improved and simplified by removing gender from the scoring system. PMID- 12625665 TI - A 2 1/2-year-old girl with reflex sympathetic dystrophy syndrome (CRPS type I): case report. AB - An unusual case of a 2 1/2-year-old girl with reflex sympathetic dystrophy (RSD) of the left arm is described. She is the youngest RSD case ever presented in the literature. Upper extremity involvement is also rare in childhood RSD. She had both physical and psychological trauma in an earthquake preceding the disease. The association of RSD with a psychological disorder is stressed and awareness of the condition to the general paediatrician is recommended for early diagnosis and successful treatment. PMID- 12625666 TI - Clinically significant benign pneumoperitoneum as a complication of percutaneous endoscopic gastrostomy. AB - Benign pneumoperitoneum is a complication of percutaneous endoscopic gastrostomy (PEG) feeding tube placement. The rate of occurrence is reported as up to 25%. It is usually an incidental finding that arises immediately following the procedure. We report a case of pneumoperitoneum that developed a significant time after the initial procedure. The patient developed intractable vomiting and had to be fed parenterally. The pneumoperitoneum resolved and the gastrostomy tube feed was re started. PMID- 12625667 TI - Current management of hypertensive disease in pregnancy. PMID- 12625668 TI - Methyldopa versus no drug treatment in the management of mild pre-eclampsia. AB - OBJECTIVES: To evaluate the efficacy of methyldopa in the treatment of mild pre eclampsia, to prevent its progress and to investigate its effect on the pregnancy outcomes. DESIGNS: Randomised clinical trial. SETTING: Wad Medani Hospital in the central Sudan. SUBJECTS: Seventy primigravidae with single, alive baby of 28-36 weeks gestational age suffering from true mild pre-eclampsia were enrolled. The patients were randomised in two groups, treatment group who received methyldopa 750-4000 mg/day (n=34) and a control group who received no treatment (n=36). All the (treatment and control) patients were drug followed as in-patients till the delivery, seen with their babies on the days 7, 42 after the delivery. MAIN OUTCOMES MEASURES: The outcomes examined were, rise of the diastolic blood pressure to 110 mm Hg or more, occurrence of imminent eclampsia or the eclampsia, if the maturity could be achieved, occurrence of intrauterine growth retardation, abruptio placentae, mode of delivery, birth weight, placental weight, perinatal death, Apgar score and referral of the babies to the pediatrician. RESULTS: Three out of 34 (8.8%) of the treatment group had a rise in the diastolic blood pressure of 110 mm Hg, 18/36 (50%) of the control had a rise in the diastolic blood pressure of 110 mmHg (p < 0.05). Three out of thirty four (8.8%) of the treatment group developed imminent eclampsia, while 10/36 (27.8) of the control group developed imminent eclampsia (p < 0.05). The maturity was achieved in 82.3% and 88.8% of the treatment and the control, respectively (p > 0.05). There were ten (14.2%) perinatal deaths, four of them in the treatment group, while six in the control (p > 0.05). There was no difference regarding birth weight, occurrence of intrauterine growth retardation, placental weight, mode of delivery, Apgar score, referral of the babies to the paediatrician. No patient developed eclampsia or abruptio placenta; there was no maternal death in both groups. CONCLUSION: Methyldopa can prevent the progress of the mild pre-eclampsia to severe pre-eclampsia, without affecting the maturity, birthweight or the neonatal outcomes. PMID- 12625669 TI - Re: Experience of road traffic accident victims at the Nairobi Hospital, Saidi and Kahoro. PMID- 12625670 TI - Socio-demographic and reproductive characteristics of mothers delivered at the Korle-Bu Teaching Hospital, Ghana. AB - OBJECTIVES: To measure selected socio-demographic and reproductive history characteristics of parturients at the Korle-Bu Teaching Hospital (KBTH), Accra, Ghana, and to compute the risk load. DESIGN: A non-randomised cross-sectional survey. SETTING: Korle-Bu Teaching Hospital, Accra, Ghana. SUBJECTS: Korle-Bu Teaching Hospital, a tertiary institution delivers about 11,000 women annually. From 1st November to 12th December 1994, 961 parturients were studied out of 978 delivered during the study period. Seventeen questionnaires were excluded from analysis because of errors and omissions that could not be corrected before the parturients' discharge from hospital. METHODS: The data sources were the patients' antenatal and delivery records, and a structured interviewer administered questionnaire. RESULTS: More than three percent of the subjects were less than 18 years, and 10.8% were over 35 years of age. Before the index delivery, 5.8% were grand multiparae. Eighteen per cent had never been to school. Seventeen per cent of parous subjects had experienced a perinatal death. The non educated had significantly more births. The mean birth interval was less than two years in 26.4%. Fifty percent of those who had been previously pregnant had a history of at least one induced-abortion. Only 21.0% of the 961 subjects had ever used a family planning method. The risk load was 53.0%. CONCLUSION: Analysis of the historical factors of parturients surveyed at the KBTH showed a high risk load related mainly to lack of education. Formal education of the female child and family health education of our women are recommended to reduce the high past abortion rate and risk load. Additionally, postpartum tubal ligation for those who have completed their families will further reduce the risk load. PMID- 12625672 TI - Utilisation of antenatal and maternity services by mothers seeking child welfare services in Mbeere District, Eastern Province, Kenya. AB - BACKGROUND: Utilisation of antenatal and maternity services is an important maternal health indicator. Increasing the proportion of mothers who are cared for in health facilities during pregnancy, childbirth and puerperium reduces the health risks to mothers and their children. OBJECTIVE: To determine the utilisation of antenatal and maternity services by mothers of Mbeere District bringing their children to the child welfare clinic. DESIGN: Cross-sectional, descriptive study. SETTING: Four rural health centres in Mbeere district. SUBJECTS: Two hundred mothers bringing their children aged one year and below to the child welfare clinic between September and December 2000. RESULTS: The proportion of mothers who utilised health facilities for antenatal and maternity services was 97.5% and 52%, respectively. Utilisation of health facilities for maternity services was significantly influenced by number of children and distance to health facility in that, as number of children increased, utilisation of maternity services reduced (chi2=8.99; p=0.027; df=1). Mothers living less than 5 km to a health facility utilised the services better than those living 5 km and beyond (chi2=7.57; p=0.0059; df=1). Among the reasons given by the mothers (individual respondents and through Focus Group Discussions) regarding dissatisfaction with the services offered included shortage of drugs and essential supplies, lack of commitment by staff, poor quality of food and lack of cleanliness in the health facilities. CONCLUSION: Coverage for antenatal services was high among mothers during their last pregnancy. However, only about half of the mothers interviewed utilised health facilities for maternity services (labour and delivery). The major constraints experienced by the mothers as they sought for the services (as reported by individual respondents and through Focus Group Discussions) included lack of transport, lack of money for transport and hospital fee and delay in admission to health facility once mothers report in labour. Lack of satisfaction with quality of care given could be the major demotivating factor in the use of health facilities for maternity services. PMID- 12625671 TI - Haemorheological changes during the menstrual cycle. AB - OBJECTIVE: To determine changes in some haemorheological parameters: haematocrit (Hct), Whole Blood Relative Viscosity (WBRV), Erythrocyte Sedimentation Rate (ESR) and Fibrinogen Concentration (FC) during the menstrual cycle. DESIGN: Cross sectional prospective study. SUBJECTS: Three hundred and fifty randomly selected female undergraduate students of the University of Port Harcourt, Port Harcourt, Nigeria. SETTING: Departments of Human Physiology and Anatomy, Faculty of Basic Medical Sciences, College of Health Sciences, University of Port Harcourt, Port Harcourt, Nigeria. MAIN OUTCOME MEASURES: Significant variation (p<0.05) were found in the values of all four haemorheological parameters assessed in the various phases of the menstrual cycle. RESULTS: Both haematocrit (Hct) and Whole Blood Relative Viscosity (WBRV) showed a similar trend, being highest in the ovulatory phase, followed by the luteal phase and follicular phase, and lowest in the menstrual phase. The Erythrocyte Sedimentation Rate (ESR) however, showed a pattern opposite to both Hct and WBRV, being highest in the menstrual phase followed by the luteal phase, and the follicular phase, and lowest in the ovulatory phase. The Fibrinogen Concentration (FC) was highest in the menstrual phase, followed by the ovulatory phase, the luteal phase and was lowest in the follicular phase. CONCLUSION: The study shows that significant variation could occur in some haemorheological parameters during the phases of the normal menstrual cycle. PMID- 12625673 TI - Gunshot injuries as seen at the Aga Khan Hospital, Nairobi, Kenya. AB - OBJECTIVE: To determine the prevalence, injury patterns, offender-victim characteristics, treatment and outcome of firearm-related injuries at the Aga Khan Hospital, Nairobi. DESIGN: Retrospective descriptive study. SETTING: The Aga Khan Hospital, a major private hospital in Nairobi. SUBJECTS: One hundred and seven patients who presented at the Aga Khan Hospital Accident and Emergency Department (January 1993-December 1998) with firearm injuries and were subsequently hospitalised. There were 97 males and 10 females aged four to 94 years. The mean age was 39 years. RESULTS: The peak incidence was in the 40-49 year age group. The male to female ratio was 10:1. The victim offender was a thug, thief or robber in 74.7% of cases. Law enforcement officers were responsible for 9.4% of the injuries. The Injury Severity Scores (ISS) ranged from one to 32 with mean score of 8.25. Injuries involving the extremities were the most prevalent. There were 31 major operations performed. The complication rate was 35.5%. Six (6.5%) of these patients died. CONCLUSION: Gunshot injuries cause profound morbidity and significant mortality. A wider and larger study needs to be undertaken to elucidate the true nature of firearm injuries. PMID- 12625674 TI - Re: Rheumatology in Kenya: 2000 and onwards. PMID- 12625675 TI - Pattern of cancer in Mbarara, Uganda. AB - OBJECTIVE: To determine the pattern of cancer in Mbarara region. DESIGN: Retrospective descriptive study. SETTING: Mbarara University of Science and Technology (MUST), Mbarara, southwestern Uganda. SUBJECTS: A total of 585 cancer patients diagnosed between 1995 and 1999 formed the basis of the analysis. RESULTS: The most frequent cancers observed among males in their descending order were Kaposis sarcoma (KS), stomach, Non Hodgkin Lymphoma (NHL), prostate and penis while in females cancer of cervix, breast, NHL, stomach and KS were found most frequent. CONCLUSION: This pattern was different from that of Kyadondo county, Uganda in which stomach and penila cancer were found to be rare whereas oesophageal cancer was one of the commonest cancer in Kyadondo but rare in Mbarara. These differences which are discussed in detail could be attributed to both environmental and genetic factors. PMID- 12625676 TI - Control of Schistosoma mansoni by the soapberry Endod (Phytolacca dodecandra) in Wollo, northeastern Ethiopia: post-intervention prevalence. AB - OBJECTIVES: To record the effect of Endod soap and spraying of soaked Endod suspension on the prevalence of human schistosomiasis. DESIGN: A cross-sectional epidemiological study in which pre- and post-intervention parasitological results were compared. SETTING: Kemise, Bati and Harbu towns in northeastern Ethiopia. SUBJECTS: The study subjects included all members of the five per cent households systematically selected from the three towns. RESULTS: In Kemise town, where suspension of ground Endod was sprayed on the stream containing infected snails, the prevalence of the disease was reduced from 59% to 53% and the mean intensity of infection was reduced from 239 eggs per gram (EPG) of faeces to 99 EPG (p < 0.05). In Bati town where Endod soap approach was used, the respective reduction in the prevalence and intensity of infection was from 51% to 43% and from 195 EPG to 162 EPG (p < 0.05). There was also a significant reduction of the disease in the control town probably due to the effects of praziquantel treatment and other factors. CONCLUSION: The reduction achieved in the prevalence and intensity of schistosomiasis after an intervention period of four years was limited. This observation corroborates the fact that molluscicides must always be considered as supplementary to chemotherapy in the control of schistosomiasis. Although both approaches can be used, the spraying approach appears to be simpler and more feasible because two or three times yearly application of Endod suspension would suppress snail population and reduce transmission. Nevertheless, the choice as to what approach to use must be made on the basis of community preference, and for some soap-effect of Endod would be attractive PMID- 12625677 TI - Pathogenic agents of chronic suppurative otitis media in Ilorin, Nigeria. AB - OBJECTIVE: To determine the type and pattern of antibiotic susceptibility of the pathogenic micro-organisms causing chronic suppurative otitis media (CSOM) in our environment. DESIGN: A retrospective study of ear discharges from patients presenting consecutively with chronic suppurative otitis media. SETTINGS: University of Ilorin Teaching Hospital, a major referral centre in the Middle Belt region of Nigeria. MAIN OUTCOME MEASURES: Bacterial isolates and their sensitivity patterns. SUBJECTS: Three hundred and seventy five patients aged between eight months and 70 years referred to the ear, nose, and throat clinic of The University of Ilorin Teaching Hospital were enrolled in the study. RESULTS: About 95.5% and 4.5% of the specimens were culture positive and negative respectively. The commonest bacterial aetiologic agents were Pseudomonas aeruginosa (26.0%) and Proteus spp (21.8%). Peak prevalence of 30.5% occurred among the 0-5 years age group. Seventy five per cent of isolates were gram negative bacteria. Ofloxacin produced 100% sensitivity in both gram positive and gram-negative organisms tested. Colistin, ceftazidime and cefuroxime were highly active (80%) against the gram-negative bacteria while erythromycin and cloxacillin were very effective (80%) against the gram-positive isolates. CONCLUSION: Chronic suppurative otitis media is still highly prevalent in our environment, affecting mainly children. The antibiotic susceptibility pattern of pathogenic isolates is different from those of other regions of Nigeria with increasing resistance recorded for some organisms. Hence, where possible and available, susceptibility tests should guide the management of CSOM in this environment, otherwise, ofloxacin if indicated and cloxacillin/erythromycin may provide relief and delay emergence of resistant strains. PMID- 12625678 TI - Burden of mental illness on family members, care-givers and the community. AB - OBJECTIVE: To determine the burden of mental illness in the family/caregiver and the community. DESIGN: A cross-sectional descriptive study. SETTING: Rehabilitation centres, community day centres, resettlement villages and in the community in three provinces (Harare, Bulawayo and Masvingo), Zimbabwe. SUBJECTS: A sample size comprising sixty six care-givers and 126 patients were consecutively interviewed for the study. RESULTS: The care-givers had a mean +/- s.d. age of 48.8 +/- 15.7 years. The majority were females (80.3%), married males, 76.9% (p=0.073), unemployed females, 94.3% (p=0.0004) and parents of the patients accounted for 51.5%. Many respondents believed that the cause of mental illness was witchcraft (31%), they experienced financial constraints (63.6%), and spent a lot of money on food (56.1%). Patients were younger than their caregivers, with a mean +/- s.d. age of 37.7 +/- 12.8 years and mostly on medications (91.3%). CONCLUSION: The caregivers were mainly women and unemployed, whereas patients were mostly male. Caregivers were faced with multiple problems, but they were more tolerant to patients' behaviour than the community at large. PMID- 12625679 TI - Effects of hypokinesia on cyclic nucleotides and hormonal regulation of calcium metabolism in rats. AB - OBJECTIVE: To study the dynamic of changes in the level of parathyroid hormone (PTH), calcitonin (CT), cyclic nucleotides (cAMP, cGMP) and calcium in the blood of rats, while in urine--phosphate, calcium and cyclic nucleotides. DESIGN: Laboratory based experiment. SETTING: Laboratory in the Department of Biochemistry, Faculty of Medicine, University of Nairobi, Nairobi, Kenya and in the Department of Biochemistry, Kharkov State University, Ukrain Republic. RESULTS: Correlation between the changes in the parameters of study was shown. This supports the theory about the relationship between branches of hormonal systems (cyclic nucleotides-calcium), which perform the central role in the regulation of homeostasis. The results show that daily excretion of calcium and phosphate is age dependent in rats during the pathogenesis of hypokinesia: the values are higher in control groups. Changes in calcium in the blood and phosphaturia in the experimental animals of all ages were attributed to changes in the intensive re-absorption process within the bones during the readaptation period of hypokinesia. CONCLUSION: Old rats have a higher ability to adaptation than younger experimental counterparts during hypokinesia. PMID- 12625680 TI - Rheumatic disorders in Sub-saharan Africa. AB - OBJECTIVE: To review prevalence of rheumatic disorders in Sub-saharan Africa and in the context of current medical practice in the region assess the need for service and educational provision. DATA SOURCES: Medline, (English, French). Pre Medline literature review from the 1950's (Current contents). Various conference reports including attendance at all three AFLAR (African League Against Rheumatism) congresses in the 1990's. Author's personal database. All cited references read in full. CONCLUSIONS: The evidence shows rheumatoid arthritis and systemic lupus erythematosus to be increasing in frequency in the indigenous populations of East, Central and South Africa but remaining rare in West Africans. Gout is now more prevalent than ever throughout the subcontinent. HIV has spawned a variety of previously rare spondyloarthropathies (reactive arthritis, psoriatic arthritis, enthesopathy) and changed the epidemiology of pyomyositis and osteomyelitis. Osteoarthritis is a universal problem. Juvenile chronic arthritis is not rare and rheumatic fever is common. Acute and chronic locomotor problems associated with diverse entities such as leprosy, brucellosis, meningococcus, alpha viruses, parasites, fluorosis, rickets and haemoglobinopathies enhance diagnostic diversity and therapeutic and educational requirements. Suggestions made to address the challenge posed by the burden of rheumatic disorders. PMID- 12625681 TI - Size, course, distribution and anomalies of the middle cerebral artery in adult Nigerians. AB - OBJECTIVE: To determine the size, course, distribution and anomalies of the middle cerebral artery in adult Nigerians. DESIGN: A retrospective study. SETTING: Department of pathology, University College Hospital, Ibadan between April and September 1999. SUBJECTS: One hundred middle cerebral arteries from patients' asymptomatic for central nervous system lesions, obtained at autopsy from fifty adult Nigerians were examined. RESULTS: The MCA arose as the wider of the two terminal branches of the internal carotid artery (ICA). Its mean diameter was 3.49 mm (CI = 3.39-3.59 mm), while the mean pre-division length was 15.43 mm (CI = 14.59-16.27 mm). In 92% of specimens, the first branches were perforators. The early branches were destined solely to the temporal lobe pole in most hemispheres (85%). In most specimens, (81%) the main trunk bifurcated while in 13% it trifurcated. One accessory MCA, and the early branches were given off before the perforators in two middle cerebral arteries, making an incidence of anomalies of 3%. There was no aneurysm. CONCLUSION: These results are similar to what has been described in the literature for Caucasians. It buttresses the assertion that anatomical anomalies of the MCA are rare. The rarity at autopsy of aneurysms of the MCA in asymptomatic Nigerian Africans is not explained by the gross morphology of this vessel. PMID- 12625682 TI - Multiple supernumerary teeth in association with malocclusion: report of two cases. AB - Although multiple supernumerary teeth without any associated syndromes are rare, their occurrence can create a variety of clinical problems such as derangement of the occlusion, prevention of eruption of permanent teeth, damage to adjacent teeth, cystic degeneration and root resorption. Hence, clinical and radiographic evaluation of patients should always be thorough in order to detect their presence. Furthermore, because the clinical management of multiple supernumerary teeth poses a great challenge to clinicians, timely, appropriate consultation and interdisciplinary approach to treatment is extremely important. We report two cases, a 14 year-old boy with eight and a 13-year-old girl with seven supernumerary teeth not associated with syndromes. In the boy, the teeth were bilaterally distributed in all quadrants in the premolar regions, and in the girl they were distributed bilaterally in the premolar regions in the mandible and bilaterally distal to the upper third molars. The clinical implications and management are discussed. PMID- 12625683 TI - The role of surgery in the management of peptic ulcer disease in the last three decades. PMID- 12625684 TI - Surgical management of peptic ulcer disease. AB - OBJECTIVE: To determine the pattern of surgically managed peptic ulcer disease. DESIGN: A retrospective study. SETTING: Department of Surgery, Moi Teaching and Referral Hospital, Eldoret, Kenya. SUBJECTS: Fifty three patients operated on for peptic ulcer disease. RESULTS: The mean age was 47 years with a male/female ratio of 1.7:1. Duodenal ulcer associated complications were the commonest with duodenal ulcer/gastric ulcer ratio of 11.5:1. Most patients had chronic peptic ulcer symptoms with inadequate or no medical treatment. Perforations were the commonest complications (56.6%) followed by gastric outlet obstruction (34.0%). Closure with omental patch was done in 83.3% of perforations while truncal vagotomy and drainage was done in gastric outlet obstruction. Hypostatic pneumonia was the commonest post-operative complication. Seventy one point seven per cent of the patients were free of dyspeptic symptoms during the brief follow up period. CONCLUSION: Most patients with chronic peptic ulcers had had inadequate treatment and perforation was the most common complication. Repair of perforations with omental patch, and truncal vagotomy and drainage procedure for gastric outlet obstruction, were satisfactory surgical methods offered to patients at the Moi Teaching and Referral Hospital. PMID- 12625685 TI - Diagnostic peritoneal lavage in the evaluation of abdominal trauma using the dipstick. AB - OBJECTIVE: To determine the accuracy and sensitivity of diagnostic peritoneal lavage in the assessment of intra-abdominal injury using the dipstick method. DESIGN: Prospective study, involving the performance of diagnostic peritoneal lavage in the out patient department and surgical wards prior to surgical intervention. SETTING: Kenyatta National Hospital-General Surgical and Orthopaedic wards and outpatient department. The study was conducted over a duration of six months, starting from January 1995 to July 1995. RESULTS: Ninety six patients with penetrating (68) and blunt (28) abdominal trauma underwent diagnostic peritoneal lavage as evaluation of the severity of abdominal trauma. Dipstick (combur 9 strips) was used to evaluate lavage effluent for red blood cells, white blood cells, protein and bilirubin. Forty three patients had positive diagnostic peritoneal lavage (DPL) results, of which 40 (93%) had positive findings at laparatomy and three (7%) had negative findings at laparatomy. The remaining 53 patients had negative DPL results and were managed conservatively. One patient with a negative DPL result became symptomatic and had a positive laparatomy. Conservatively managed patients were discharged after 24 hours observations without any complications. DPL had an accuracy and sensitivity of 93% and specificity of 98%. CONCLUSION: Diagnostic peritoneal lavage is a cheap, safe and reliable method for assessment of abdominal trauma. The method is easy to perform by trained junior doctors in the OPD, or as a bedside procedure. Use of this method reduced negative laparotomy rate from 50% to 6.9% and average duration of stay from 6.5 days to 1.9 days. This method is recommended as a basic tool in the assessment of abdominal trauma patients. PMID- 12625687 TI - Cancer of the cervix: knowledge and attitudes of female patients admitted at Muhimbili National Hospital, Dar es Salaam. AB - OBJECTIVES: To determine the level of knowledge of basic symptoms of cancer of the cervix among Tanzanian females and to determine causes of late presentation with advanced disease among cancer patients. DESIGN: Hospital based cross sectional study. SETTING: Muhimbili National Hospital, Dar es Salaam, Tanzania. SUBJECTS: Eighty nine cervical cancer patients and 178 controls were interviewed between August 1999 and January 2000. Data was analyzed using Epi-Info version 6.04. RESULTS: At Muhimbili National Hospital most patients are admitted in very advanced stages of the disease (Stage IIb and IV). We determined, using a structured questionnaire, knowledge of basic symptoms of cancer of the cervix, attitude and reasons for late presentation among female patients admitted at Muhimbili National Hospital gynaecological ward. The mean age of cases was 48.8(SD11.1) years and the mean parity was 6.7 years were comparable to that of control, which were 45 years(SD10.8) and mean parity of 6.6 respectively. Mean age in years at marriage was lower for cases 17.5(SD 2.9) than controls 18.8(SD3.5). Majority of cases (50.6%) and controls (23.6%) were illiterate, and 21.3% of cases and 33.7% of controls had incomplete primary education. Majority of both cases (47.23%) and controls (56.7%) had no routine gynaecological examination and they did not find it necessary. More than 90% of the cases were in advanced stages of the disease (stage IIb-IV). CONCLUSION: Both cases and controls had low knowledge of basic symptoms of cancer of the cervix and as a result most of those who happen to have problems reported late with advanced disease. PMID- 12625686 TI - Anaemia in women of reproductive age in Dar-es-Salaam, Tanzania. AB - BACKGROUND: Anaemia is among the greatest health problems in reproductive age women in developing countries. OBJECTIVES: To estimate the prevalence of anaemia among non-pregnant parous women, and to investigate the main underlying cause for the anaemia. SETTING: A sub-urban Maternal and Child Health Clinic (MCH) in Dar es Salaam. DESIGN: Cross-sectional. METHODS: Consecutive parous non-pregnant women who had brought their children for vaccination and/or had come for family planning to Mbagala MCH clinic were invited to participate in the study. Obstetric and social history was recorded, and their height and weight were checked. Haemoglobin was measured using HemoCue hemoglobinometer. Anaemic women were further investigated to determine the cause of anaemia by haematological and biochemical tests. RESULTS: Five hundred and four parous non-pregnant women were screened, 49% were anaemic (Hb <12 g/dl) and 1.6% severely anaemic (Hb <7 g/dl). Anaemia was not related to socio-demographic and obstetric history characteristics, but decreased significantly with increasing Body Mass Index (BMI) (p=0.042). The prevalence of anaemia was significantly lower in women using hormonal contraceptives, compared to non-users (36% vs 54%) (p=0.04). Eighty seven percent of the anaemic women were iron deficient and 8.7% had elevated serum C-reactive protein indicating undiagnosed infections. CONCLUSION: Nutritional deficiencies in women have to be corrected before and between pregnancies and all contacts women have with the health system should be utilised for anaemia control interventions, in addition to long-term community approaches. To improve maternal health calls for a broader agenda and a change of approach in the MCH-clinics. PMID- 12625688 TI - Chlamydia as a cause of late neonatal pneumonia at Kenyatta National Hospital, Nairobi. AB - BACKGROUND: Neonatal pneumonia is a common cause of morbidity and mortality all over the world. The problem is known to be higher in resource poor third world countries. Organisms (such as chlamydia) not covered by routine laboratory tests and regular antibiotic regimes may frequently contribute towards the causation of late neonatal pneumonia. It is therefore useful to gather epidemiological evidence to guide in the routine diagnosis and treatment of such infections. OBJECTIVE: To determine the prevalence of chlamydia associated pneumonia among infants developing the disease between the 7th and 30th days of life (late neonatal pneumonia). DESIGN: Cross sectional survey. SETTING: Newborn Unit, Kenyatta National Hospital. SUBJECTS: Fifty two newborns clinically diagnosed as having late neonatal pneumonia. They were all subjected to chest X-rays to confirm the clinical diagnosis. Nasopharyngeal aspirates for chlamydia antigen detection tests were then performed on all patients. The study was undertaken during the months of September through to November 2000. MAIN OUTCOME MEASURES: The proportion of newborns with late neonatal pneumonia that have chlamydia trachomantis as the sole or contributory causative agent. RESULTS: Fifty two newborns of postnatal age between seven and 30 days were recruited. Their sex distribution was about 1:1. Thirty three (63.5%) of these infants were found with chlamydia in their upper airways. Thirty out of 47 available chest X-rays, representing 63.8% had evidence of interstitial pneumonitis. Chlamydia associated pneumonia indicated by the presence of both interstitial pneumonia and colonization of the upper air ways was present in 24 out of 47 patients, 51% of the total cases of late neonatal pneumonia. When X-rays alone were compared with our gold standard for the diagnosis of chlamydia pneumonia (radiology and colonization), we computed a sensitivity of 100%, specificity 73%, negative predictive value 100% and positive predictive value of 80%. Mode of delivery, birth weight and gestation had no association with nasopharyngeal colonization by chlamydia or actual diagnosis of chlamydia pneumonia. CONCLUSION: The prevalence of chlamydia associated infection among newborns with late neonatal pneumonia at Kenyatta National Hospital is 51%, eight times more than that reported elsewhere. Chest X-rays appear to be a reliable diagnostic tool in this group. The use of antichlamydial drugs in addition to the regular antibiotics whenever a diagnosis of late neonatal pneumonia is made is justifiable. PMID- 12625689 TI - Molecular markers in epidemiological monitoring of the spread of resistance to antimalarials: a review. AB - OBJECTIVE: To review the prevalence and distribution of resistance to chloroquine and pyrimethamine-sulphadoxine combination and the use of molecular markers for monitoring the spread of the resistance. DATA SOURCES: Literature search on compact disk-read only memory (CD-ROM), Medline and Internet, using the key words: Malaria and epidemiology, malaria and resistance, sulphadoxine pyrimethamine resistance and chloroquine resistance. Some articles were manually reviewed. STUDY SELECTION: Relevant studies or articles on resistance to chloroquine, sulphadoxine pyrimethamine combination and other antimalarials and molecular resistance markers from various sources are included in the review. DATA EXTRACTION: From individual study or articles. DATA SYNTHESIS: Information on antimalarial resistance is harmonised under the headings; Introduction, Prevalence and distribution of resistance to antimalarials, Use of molecular markers for epidemiological monitoring of antimalarial resistance. CONCLUSION: The spread and status of resistance to sulphadoxine-pyrimethamine (SP) and chloroquine should be monitored constantly in major health facilities. This will not only detect the emergence of resistance to these drugs but also generate information on the extent of resistance to these antimalarials. Mutations in the dhfr and dhps genes can be used as markers in SP resistance surveilance while the presence of pfcrt mutations thought to confer resistance should also be analysed to ascertain whether they truly correlate to the resistance patterns that have been observed in various malarious regions. Little is known on the interaction and exact role(s) of PfCRT protein in conferring the resistance trait. PMID- 12625690 TI - Impact of cattle keeping on human biting rate of anopheline mosquitoes and malaria transmission around Ziway, Ethiopia. AB - OBJECTIVE: To assess the impact of livestock keeping on the human biting rate (HBR) of anopheline mosquitoes and malaria transmission around Ziway in the middle course of the Ethiopian Rift Valley. DESIGN: As a passive experiment, man landing captures were done in homesteads with mixed dwelling, separate cattle shed and without livestock; and as an active experiment, captures were in experimental tukuls (huts) of cattle, goats, and without livestock. Parasite and spleen rates of children were compared among those residents under variable living conditions mentioned for passive experiment. SUBJECTS: For entomological study, human-baits were used for man-landing captures of mosquitoes. Study subjects for parasitological and clinical studies were children below 10 years old. MAIN OUTCOME MEASURES: Human-biting rate (HBR) of anopheline mosquitoes; and the parasite and spleen rates of the study subjects in different living conditions. RESULTS: In the passive experiment, the mean HBR of Anopheles arabiensis in mixed dwelling, separate cattle shed and without livestock was 8.45, 4.64 and 5.97, respectively. Similarly, the HBR of An. pharoensis was 2.88, 1.79 and 1.61, respectively. In the active experiment, the mean HBR of An. arabiensis in tukuls with cattle, goats, and without livestock was 3.50, 3.38 and 1.43 respectively; while that of An. pharoensis was 0.37, 0.70 and 0.55 respectively. Parasitologically, mean parasite rates of 26.67%, 15.05% and 23.85% were, respectively, recorded from children living under the above conditions stated for passive experiment. Similarly, the mean spleen rates of 50.0%, 26.9%, and 47.37% were recorded, respectively. CONCLUSION: These observations in the present study indicate that the presence of cattle in homesteads tends to increase the man biting rate of An. arabiensis, although differences in the mean HBR of vector mosquitoes were not statistically significant for all groups. In contrast, cattle keeping in separate cattle sheds outside of the human dwellings tends to reduce the man biting rate of An. arabiensis and malaria transmission in the study area. PMID- 12625691 TI - Validity of random blood glucose as a predictor of the quality of glycaemic control by glycated haemoglobin in out-patient diabetic patients at Kenyatta National Hospital. AB - BACKGROUND: Patients with diabetes mellitus in Kenya come to the hospital for follow-up visits very infrequently. For most of these patients their blood glucose monitoring is done only on the day of visit to the doctor. OBJECTIVE: To determine how well the physician-based morning random blood level determines or reflects the quality of glycaemic control. DESIGN: Cross-sectional study (morning, random blood glucose taken between 8.00 a.m. and 12.00 noon). SETTING: Out-patient diabetic clinic of Kenyatta National Hospital. SUBJECTS: Patients with diabetes mellitus either type 1 or type 2 attending the out-patient clinic. MAIN OUTCOME MEASURES: Random blood glucose (morning) and glycated haemoglobin (HbA1c). RESULTS: The morning random glucose level had a linear relationship with glycated haemoglobin levels taken simultaneously. A blood glucose level of 7 mmol/l had 92.7% sensitivity for good control (HbA1c < or = 7.8%) on a blood sample which was taken simultaneously and 59.8% specific for the same. When blood glucose cut-off level was raised to 10 mmol/l sensitivity fell to 66.3% for HbA1c < or = 7.8%, and 83.2% specificity for poor glycaemic control (HbA1c > 7.8%). There was marked fall in sensitivity of rising random blood glucose level in predicting good glycaemic control in our study, with concomitant rise in specificity of those high cut-off levels of blood glucose in predicting poor glycaemic control. CONCLUSION: Morning random blood glucose in the ambulatory diabetic patients related well to simultaneously assayed HbA1c. Blood glucose within usual therapeutic targets of 4-8 mmol/l predicted good glycaemic control (HbA1c < or = 7.8%) with high sensitivity at the range of 86.3-98.4%. In resource poor settings, the morning random blood glucose assay, which is done in patients who may attend the diabetic clinic in the morning hours, may be used to predict the quality of their diabetic control. However caution should be exercised in its widespread use because its overall applicability may be clinic-specific depending largely on the average metabolic control of the diabetic population using that clinic. Further studies need to be done to relate HbA1c to blood glucose levels obtained at different times of the day in this population to determine the best predictor of good glycaemic control. PMID- 12625692 TI - Decision-intervention interval in ruptured uteri in Ile-Ife, Nigeria. AB - OBJECTIVE: To determine the decision-intervention interval in ruptured uterus with a view of overhauling the management strategy thereby improving the maternal and perinatal outcome. DESIGN: Cross-sectional survey based on secondary data done between 1990-1999. SETTING: Obafemi Awolowo University Teaching Hospital Complex, Ile-Ife, Nigeria. SUBJECTS: One hundred and two consecutive women who had uterine rupture. RESULTS: The decision-surgical intervention interval ranged from 30 minutes to 4.5 hours. The major reason for delay was unavailability of compatible blood (88.2%), followed by lack of electricity (4.9%), unsterile instruments (3.9%), waiting for ambulance to get senior obstetricians (2.9%), delay in arrival of anaesthetist (1.9%) and neonatologists (1.9%). The peri-natal mortality rate (PNMR) was 843 per 1000 total births and maternal mortality rate (MMR) 4902 per 100,000 births. CONCLUSION: The maternal and perinatal outcome in uterine rupture would be improved by early diagnosis and avoidance of preoperative delay through availability of essential obstetric services. PMID- 12625693 TI - Cancrum oris in HIV infected children in Lesotho: report of two cases. AB - Cancrum Oris (Noma) has been known to mankind since time immemorial. Its public health importance is lately being realised especially in Africa, where majority of the cases occur. It affects those whose immune systems are compromised and thus, it is no wonder that it is being reported in HIV infected individuals. This development could have serious implications regarding the impact of the HIV pandemic globally and in particular the African region where poverty is rife. PMID- 12625694 TI - Goldenhar's syndrome: case report. AB - A case of Goldenhar's Syndrome (oculoauriculovertebral dysplasia) in a 3-day-old Nigerian neonate with right anophthalmos and lipodermoid, left limbal dermoid, bilateral preauricular appendages and mandibular hypoplasia is presented. The lipodermoid was attached to the tarsal conjunctiva of the lower lid of the anophthalmic side. The causative factor was presumably maternal drug (traditional medicine) ingestion at three months gestation. No similar case has been reported previously. The literature on Goldenhar's Syndrome is briefly reviewed. PMID- 12625695 TI - Convergent solid-phase synthesis of symmetrical benzo[b]furan's dimerizer. PMID- 12625696 TI - Formamide as a combined ammonia synthon and carbon monoxide source in fast palladium-catalyzed aminocarbonylations of aryl halides. PMID- 12625697 TI - Ion-extraction ladder sequencing from bead-based libraries. AB - Ion-extraction mass spectrometry of ladders of mixtures of isotopically labeled compounds from single beads allows the unambiguous sequencing of bead-based peptides and offers significant advantages over traditional methods of library analysis. PMID- 12625698 TI - Solid-phase combinatorial library of norstatine-type isosters by the nitroaldol reaction. AB - A combinatorial library of norstatine-type peptide isosters as putative inhibitors of aspartic proteases is presented. The library was synthesized using a split-and-mix strategy designed to afford a one-bead-two-compounds library with the isosteric elements positioned centrally in peptide chains. Application of ladder synthesis during library generation enabled structure identification by MALDI-TOF mass spectroscopy. The library was screened against aspartic protease renin, and two types of inhibitors were identified, that is, XXX-psi[CHRCHOH)-XXX and an aldehyde arising from unreacted starting material. Selected hits were resynthesized and assayed in solution, revealing inhibitors of nanomolar potency. PMID- 12625700 TI - High-throughput manual parallel synthesis using SynPhase crowns and lanterns. AB - The high-throughput manual solid-phase parallel synthesis of libraries comprising thousands of discrete samples using pellicular supports (i.e. SynPhase crowns and lanterns) and a suite of novel tools and techniques is described. Key aspects of this approach include the combination of a split-split-split synthesis strategy with spatial encoding to differentiate thousands of crowns, the rapid washing and filtration of up to 48 reaction vessels in parallel, the application of an inexpensive and environmentally friendly technique to remove trifluoroacetic acid from sixteen 96-well plates in parallel, and a high-throughput method for removing cleaved crowns from reusable pin racks. Tens of thousands of discrete samples have been produced in-house using this conceptually and operationally straightforward strategy. PMID- 12625699 TI - Imaging combinatorial libraries by mass spectrometry: from peptide to organic supported syntheses. AB - Supported peptide and drug-like organic molecule libraries were profiled in single nondestructive imaging static secondary ion mass spectrometric experiments. The selective rupture of the bond linking the compound and the insoluble polymeric support (resin) produced ions that were characteristic of the anchored molecules, thus allowing unambiguous resin bead assignment. Very high sensitivity and specificity were obtained with such a direct analytical method, which avoids the chemical release of the molecules from the support. Libraries issued from either mix-and-split or parallel solid-phase organic syntheses were profiled, demonstrating the usefulness of such a technique for characterization and optimization during combinatorial library development. Moreover, the fact that the control was effected at the bead level whatever the structure and quantity of the anchored molecules allows the sole identification of active beads selected from on-bead screening. Under such circumstances, the time-consuming whole-library characterization could thus be suppressed, enhancing the throughput of the analytical process. PMID- 12625701 TI - Synthesis of 4-(5-iodo-3-methylpyrazolyl) phenylsulfonamide and its elaboration to a COX II inhibitor library by solution-phase suzuki coupling using Pd/C as a solid-supported catalyst. AB - An effective synthesis of 4-(5-iodo-3-methylpyrazolyl) phenylsulfonamide has been developed. This aromatic iodide template served as an efficient oxidative addition partner for the preparation of a solution-phase library of Celecoxib analogues via Suzuki coupling using Pd/C, a readily filterable catalyst. PMID- 12625702 TI - High-throughput one-bead-one-compound approach to peptide-encoded combinatorial libraries: MALDI-MS analysis of single TentaGel beads. AB - The identification of pharmacologically promising compounds (lead compounds) from combinatorial libraries is frequently limited by the throughput of the analytical technique employed. Fourier transform mass spectrometry (FTMS) offers high sensitivity, mass accuracy (m/Deltam > 500 000), and sequencing capabilities. A rapid and efficient method for high-throughput analysis of single beads from peptide-encoded combinatorial libraries with matrix-assisted laser desorption/ionization (MALDI) mass spectrometry is presented. Encoding peptides on single beads are identified and structurally characterized by MALDI time-of flight (TOF) and ultrahigh-resolution MALDI Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry. A strategy of on-probe sample preparation is developed to minimize handling of the beads. PMID- 12625703 TI - Efficient synthesis of sulfonic, phosphoric, and phosphinic esters employing alkylating polymer-bound reagents. AB - The efficient esterification of various sulfonic acids and sulfonates using polymer-bound triazenes based on the triazene T2 linker is described. Esterification of enantiopure alpha-substituted sodium sulfonates was performed in the presence of an alkylating resin without racemization. Racemization is a serious drawback in the esterification route via sulfonyl chlorides because of intermediate sulfene formation. To demonstrate the versatility of this protocol, phosphoric and phosphinic acids have been converted into the corresponding esters as well. All products were obtained in good yield and excellent purities without any further purification steps. PMID- 12625704 TI - Preparation of thioamide building blocks via microwave-promoted three-component kindler reactions. AB - A microwave-enhanced variation of the Kindler thioamide synthesis is introduced. Taking advantage of the sealed vessel capabilities of a dedicated single-mode microwave reacto,r a diverse selection of 13 aldehyde and 12 amine precursors was utilized in the construction of a representative 34-member library of substituted thioamides. The three-component condensations of aldehydes, amines, and elemental sulfur were carried out using 1-methyl-2-pyrrolidone (NMP) as solvent employing microwave flash heating at 110-180 degrees C for 2-20 min. A simple workup protocol allows the isolation of synthetically valuable primary, secondary, and tertiary thioamide building blocks in 83% average yield and >90% purity. PMID- 12625705 TI - Method for analysis of polymer-supported organic compounds using mass spectrometry direct insertion. AB - A new approach on the use of mass spectrometry direct-insertion and a quadrupole detector for analysis of organic compounds supported in solid phase has been developed. This is a simple and efficient method based on cleavage due to the thermal-instability of the benzylic group of most commercial resins. The cleavage of supported compounds takes place in the spectrometer as a consequence of the high temperature in the instrument's chamber. These compounds are detected using a similar fragmentation pattern and a molecular ion corresponding to the same compound obtained by traditional synthesis. Polymer degradation fragments do not interfere with the spectrum interpretation, because only a few peaks and low intensities are detected. We report here the identification of different types of compounds supported in Merrifield resin, such as bis-o-aminobenzamides and simple aromatic and aliphatic compounds, using this new approach. PMID- 12625706 TI - Parallel solid-phase synthesis of 2-imino-4-oxo-1,3,5-triazino[1,2 a]benzimidazoles via tandem aza-Wittig/heterocumulene-mediated annulation reaction. AB - The parallel synthesis of a large number of 2-imino-4-oxo-1,3,5-triazino[1,2 a]benzimidazole derivatives via a solid-phase 1,3,5-triazino-annulation reaction is described. The solid-phase approach involves the in situ generation of iminophosphorane derivatives derived from resin-bound 2-aminobenzimidazoles employing Mitsunobu conditions. The subsequent Aza-Wittig reaction of the iminophosphoranes with isocyanates leads to highly reactive carbodiimides, which undergo an intramolecular heterocyclization reaction to form tetrasubstituted 2 imino-4-oxo-1,3,5-triazino[1,2-a]benzimidazoles in high yields (74-94%) and good purity (>80%). PMID- 12625707 TI - Recycling and reuse of a polymer-supported scavenger for amine sequestration. AB - The regeneration and reuse of a supported scavenger 1 for amine sequestration has been achieved up to three times without significant loss of activity. The scavenging process between the aldehyde resin 1 and a range of amines has been investigated in detail to determine the scope of this scavenger. Its application for the rapid purification of a small library of secondary amines 7a-j has also been demonstrated, and it has been shown that the large excess of scavenger resin used can be recovered and recycled, making this a more cost-effective process. PMID- 12625708 TI - Synthesis of tetrahydro-1,4-benzodiazepine-2-ones on hydrophilic polyamide SynPhase lanterns. AB - Solid-phase synthesis is greatly dependent on the solid support. Here, we report the use of a new hydrophilic grafted surface on SynPhase lanterns in solid-phase organic chemistry. A convenient and facile solid-phase synthesis of disubstituted 1,4-benzodiazepine-2-ones on polyamide SynPhase lanterns is described. The key step of the synthesis involved a reduction-cyclization of a nitroaryl methyl ester with a mixture of tin(II) chloride dihydrate and ammonium acetate in water and ethanol at elevated temperature to give the desired target compounds. A library of 21 disubstituted 1,4-benzodiazepine-2-ones was prepared. PMID- 12625709 TI - Combinatorial lead optimization of [1,2]-diamines based on ethambutol as potential antituberculosis preclinical candidates. AB - Despite relatively modest potency, ethambutol (EMB, (S,S)-[N,N-di-2-amino-1 butanol]ethylenediamine) is a mainstay of contemporary chemotherapy for the treatment of tuberculosis. We have developed a solid-phase synthesis of 1,2 diamine analogues of EMB using a novel acylation-reduction sequence that is compatible with high-throughput 96-well format chemistry. Using this procedure, we have synthesized 63 238 diamine analogues in pools of 10 that are suitable for testing. MIC and a target-based reporter assay were used to direct deconvolution of 2796 individual compounds from these mixtures, and the 69 most potent molecules were resynthesized in milligram quantities for hit confirmation. Purification of these individual active diamine analogues allowed the identification of 26 compounds with activity equal to or greater than EMB. Amines which occurred most frequently in active compounds included many with large hydrophobic moieties, suggesting that optimization was perhaps selecting for the isoprenoid binding site of the arabinosyltransferase target of EMB. N-Geranyl-N' (2-adamantyl)ethane-1,2-diamine (109), the most active of these diamines, displayed a 14-35-fold improvement in activity in vitro against Mycobacterium tuberculosis, as compared to EMB. PMID- 12625710 TI - Solid-phase rhodium carbenoid N-H insertion reactions: the synthesis of a diverse array of indoles. AB - A solid-phase synthesis of an array of indoles is reported. The key step in our approach involves a N-H insertion reaction of N-alkylanilines into a highly reactive polymer-bound rhodium carbenoid intermediate to yield the corresponding alpha-arylamino-beta-ketoester. These insertion products were then treated under acid-catalyzed cyclodehydration conditions to yield a series of polymer-bound indole esters, which were subsequently cleaved from the resin under Lewis acid promoted amidation conditions to yield the desired indoles in good yields and with excellent purities. PMID- 12625711 TI - Poly(3-hydroxyalkanoate)s functionalized with carboxylic acid groups in the side chain. AB - Biodegradable polyesters represent an important class of materials, and one subset of these polymers are the bacterially produced poly(3-hydroxyalkanoate)s (PHA), a bacterially produced material. These polymers are very hydrophobic, and chemical methods to increase their hydrophilicity will ultimately lead to new applications. Many copolymers of PHA are known that contain simple, nonpolar functionality in the side chain, and we explored the conversion of side-chain olefins to carboxylic acids under conditions that minimize molecular weight degradation. With the use of osmium tetraoxide and oxone, the conversion proceeded to completion with little backbone degradation, which was confirmed with NMR, IR, and gel permeation chromatography (GPC). The solubility character of the polymer before and after reaction is very different, and several solvents were explored including acetone, tetrahydrofuran (THF), and water. PMID- 12625712 TI - Sustainable enzymatic preparation of polyaspartate using a bacterial protease. AB - Diethyl l-aspartate was polymerized by a bacterial protease from Bacillus subtilis (BS) in organic solvent at a temperature between 30 and 50 degrees C to yield alpha-linked poly(ethyl l-aspartate) having an M(w) of up to 3700 and a maximum polymer yield of 85%. The best polymerization conditions were the 40 degrees C polymerization of diethyl l-aspartate using 30% protease BS containing 4.5 vol % water in acetonitrile for 2 days. Poly(ethyl l-aspartate) was readily depolymerized by the enzyme into the oligomeric and monomeric l-aspartate in aqueous acetonitrile. Poly(sodium aspartate) prepared by the saponification of poly(ethyl l-aspartate) was readily biodegradable by activated sludge obtained from the municipal sewage treatment plant. Also, poly(sodium aspartate) was depolymerized by the hydrolase enzyme into the monomeric aspartate. These results may indicate the sustainable chemical recycling and biorecycling of this polymer. PMID- 12625713 TI - Solid-phase handling of hydrophobins: immobilized hydrophobins as a new tool to study lipases. AB - Hydrophobins are fungal proteins that self-assemble spontaneously at hydrophilic hydrophobic interfaces and change the polar nature of the surfaces to which they attach. This attribute can be used to introduce hydrophobic foci on the surface of hydrophilic supports where hydrophobins are attached by covalent binding. In this paper, we report the binding of Pleurotus ostreatus hydrophobins to a hydrophilic matrix (agarose) to construct a support for noncovalent immobilization and activation of lipases from Candida antarctica, Humicola lanuginosa, and Pseudomonas flourescens. Lipase immobilization on agarose-bound hydrophobins proceeded at very low ionic strength and resulted in increased lipase activity and stability. The enzyme could be desorbed from the support using moderate concentrations of Triton X-100, and its enantioselectivity was similar to that of lipases interfacially immobilized on conventional hydrophobic supports. These results suggest that lipase adsorption on hydrophobins follows an "interfacial activation" mechanism; immobilization on hydrophobins offers new possibilities for lipase study and modulation and reveals a new application for fungal hydrophobins. PMID- 12625714 TI - Enzymatic synthesis and curing of biodegradable epoxide-containing polyesters from renewable resources. AB - Epoxide-containing polyesters were enzymatically synthesized via two routes using unsaturated fatty acids as starting substrate. Lipase catalysis was used for both polycondensation and epoxidation of the unsaturated fatty acid group. One route was synthesis of aliphatic polyesters containing an unsaturated group in the side chain from divinyl sebacate, glycerol, and the unsaturated fatty acids, followed by an epoxidation of the unsaturated fatty acid moiety in the side chain of the resulting polymer. In another route, epoxidized fatty acids were prepared from the unsaturated fatty acids and hydrogen peroxide in the presence of lipase catalyst, and subsequently the epoxidized fatty acids were polymerized with divinyl sebacate and glycerol. The polymer structure was confirmed by NMR and IR, and for both routes, the high epoxidized ratio was achieved. Curing of the resulting polymers proceeded thermally, yielding transparent polymeric films with high gloss surface. Pencil scratch hardness of the present films improved, compared with that of the cured film obtained from the polyester having an unsaturated fatty acid in the side chain. The obtained film showed good biodegradability, evaluated by BOD measurement in an activated sludge. PMID- 12625715 TI - Characterization of Polylactide-b-polyisoprene-b-polylactide Thermoplastic Elastomers. AB - Model polylactide-b-polyisoprene-b-polylactide (PLA-PI-PLA) triblock copolymers were prepared by an efficient protocol starting with alpha,omega-dihydroxy polyisoprene (HO-PI-OH). Using a moderately electrophilic Al(O-i-Pr)(3) catalyst and carefully controlling the ratio of Al to HO-PI-OH avoided gel formation and resulted in acceptable lactide polymerization rates. The triblock copolymers were free of homopolymer or diblock contaminants as determined by chromatographic and spectroscopic analyses. Three representative PLA-PI-PLA materials were prepared with spherical, cylindrical, and lamellar morphologies as confirmed by small angle X-ray scattering and transmission electron microscopy. We employed dynamic mechanical analysis and tensile testing to assess the viscoelastic and mechanical behavior. The morphology largely determined the tensile properties of these materials, with the Young's modulus and ultimate tensile strength following predicted trends. Excellent elongations were achieved especially for the PLA-PI PLA sample with the cylindrical morphology, and the PLA-PI-PLA sample with the spherical morphology showed the best elastomeric recovery. Microphase alignment and pull rate significantly influenced the resultant tensile properties. PMID- 12625716 TI - A chemical surface modification of chitosan by glycoconjugates to enhance the cell-biomaterial interaction. AB - The use of wheat germ agglutinin (WGA), a lectin molecule, to modify chitosan and enhance the cell-biomaterial interaction was examined. The percentage of living fibroblast cells on the surfaces of tissue culture polystyrene (TCPS) control, WGA-modified chitosan, and unmodified chitosan films increased to 99%, 99%, and 85%, respectively, after seeding for 48 h. DNA staining revealed that a portion of fibroblasts cultivated on chitosan films( )were undergoing apoptosis. In contrast, fibroblasts growing on WGA-modified chitosan film surfaces did not show any indication of apoptosis. The number of fibroblast cells was the highest on the WGA-modified chitosan surfaces, followed by the TCPS and unmodified chitosan surfaces. This WGA-mediated enhancement on the fibroblast cell-biomaterial interaction was cell type dependent. Other types of cells may need different lectin molecules for enhanced interaction with biomaterials. Further, the evaluation of the heat shock protein (HSP) mRNA expression indicated that HSP 90 expression was increased in the fibroblast cells cultivated on chitosan films and decreased to basal levels on the WGA-modified chitosan films. Taken together, our data suggest that the use of WGA and other lectin molecules to enhance the cell biomaterial interaction via oligosaccharide-mediated cell adhesion is a promising way to improve cell adhesion and proliferation, the two key issues in tissue engineering. PMID- 12625717 TI - Polyelectrolyte Complexes: Interactions between Lignosulfonate and Chitosan. AB - The interactions between high molecular weight chitosans (fraction of acetylated units (F(A)) = 0.10 or 0.50) and lignosulfonates of varying molecular weights (5000-400000 g/mol) and degrees of sulfonation (0.39-0.64) were studied. Lignosulfonates and chitosans form primarily insoluble polyelectrolyte complexes when mixed at pH 4.5, where the polymers are oppositely charged. In contrast, no complex formation occurred at pH 8, as shown by using a chitosan with F(A) = 0.50, which is soluble at this pH. Thus, a positively charged chitosan is a prerequisite for interactions leading to insoluble complexes with lignosulfonates. It is therefore unlikely that complex formation involves the formation of covalent sulfonylamide linkages as proposed in the literature. The composition of the complexes varied to some degree with the mixing ratio and molecular weight of lignosulfonate, but in most cases compact complexes with a sulfonate/amino ratio close to 1.0 were formed, suggesting that all sulfonate groups are accessible for interactions with chitosan. The influence of the ionic strength and temperature on the complex formation and the behavior of the precipitated complexes were in agreement with that expected for classical polyelectrolyte complexes where the associative phase separation is primarily governed by the increase in entropy due to the release of counterions. PMID- 12625718 TI - On the characterization of pH-sensitive liposome/polymer complexes. AB - A randomly alkylated copolymer of N-isopropylacrylamide, methacrylic acid and N vinyl-2-pyrrolidone was characterized with regard to its pH- and temperature triggered conformational change. It was then complexed to liposomes to produce pH responsive vesicles. Light scattering and differential scanning calorimetry experiments performed at neutral pH revealed that the polymer underwent coil-to globule phase transition over a wide range of temperatures. At 37 degrees C and pH 7.4, although the polymer was water-soluble, Fourier transform infrared spectroscopy analysis showed that it was partly dehydrated. At acidic pH, the decrease in the lower critical solution temperature was accompanied by an increase in cooperativity degree of the phase transition. Complexation of copolymer to liposomes did not substantially influence its phase transition. The liposome/copolymer complexes were stable at neutral pH but rapidly released their contents under acidic conditions. The copolymer slightly increased liposome circulation time following intravenous administration to rats. The addition of poly(ethylene glycol) to the formulation had a detrimental effect on pH sensitivity but enhanced substantially the circulation time. PMID- 12625719 TI - Highly branched poly(L-lysine). AB - This paper describes the synthesis of several novel water-soluble highly branched polypeptides. The synthesis starts with the ring-opening polymerization of epsilon-benzyloxycarbonyl-l-lysine N-carboxyanhydride (Z-Lys NCA) or epsilon trifluoroacetyl-l-lysine N-carboxyanhydride (TFA-Lys NCA), followed by end functionalization of the peptide chain with N(alpha),N(epsilon)-di(9 fluorenylmethoxycarbonyl)-l-lysine (N(alpha),N(epsilon)-diFmoc Lys). Deprotection of the N(alpha),N(epsilon)-diFmoc Lys end group affords two new primary amine groups that can initiate the polymerization of a second generation of branches. Repetition of this ring-opening polymerization-end functionalization sequence affords highly branched poly(epsilon-benzyloxycarbonyl-l-lysine) (poly(Z-Lys)) and poly(epsilon-trifluoroacetyl-l-lysine) (poly(TFA-Lys)) in a small number of straightforward synthetic steps. Removal of the side-chain protective groups yields water-soluble and highly branched poly(l-lysine)s, which may be of potential interest for a variety of medical applications. PMID- 12625720 TI - Rheological properties and gelation of aqueous cellulose-NaOH solutions. AB - The shear rheology of a microcrystalline cellulose dissolved in a 9% NaOH aqueous solution was studied in the steady and oscillatory modes. The cellulose-(9% NaOH H(2)O) mixtures show not to be true solutions. In the dilute regime, with cellulose concentration below 1%, the rheological behavior is typical of the one of suspensions. The formation of cellulose aggregates is favored when temperature is increased. In the semidilute regime, an irreversible aggregate-based gelation occurs, being faster with increasing temperature. PMID- 12625721 TI - Layer-by-layer engineering of biocompatible, decomposable core-shell structures. AB - The objective of the present investigation was to fabricate composite colloidal particles consisting of a sacrificial, decomposable template of biodegradable nature covered with biocompatible polyelectrolyte multilayers using the layer-by layer sequential adsorption technique. Poly-dl-lactic acid and poly(dl-lactic-co glycolic acid) were chosen to design the microparticulate template, and a preliminary feasibility study was carried out with poly(styrene sulfonate sodium) poly(allylamine hydrochloride) as shell components. The properties of both core shell and hollow structures obtained by core dissolution were characterized by confocal laser scanning microscopy, microelectrophoresis, scanning force microscopy, and scanning electron microscopy. The concept was then extended to biocompatible polyelectrolytes as shell wall building blocks to deduce stable hollow capsules with tailored properties. Uniform, complete coating with oppositely charged polyelectrolyte pairs was achieved for all the combinations investigated. The results demonstrate that polyester microparticles could serve as viable alternative components to conventionally employed templates to derive hollow capsules with defined size, shape, and shell thickness. With all the components used for fabrication being biocompatible, these polyelectrolyte capsules may find interesting applications in the fields of biology, biochemistry, biotechnology, and drug delivery. PMID- 12625722 TI - Ionic strength dependence of protein-polyelectrolyte interactions. AB - The effect of univalent electrolyte concentration on protein-polyelectrolyte complex formation has been measured by frontal analysis continuous capillary electrophoresis (FACCE) and turbidimetry for the interaction of bovine serum albumin (BSA) with a synthetic hydrophobically modified polyacid, for BSA with (porcine mucosal) heparin (Hp), a highly charged polyanion, and for Hp and insulin. All three highly diverse systems display maxima or plateaus in complex formation in the range of ionic strength 5 < I < 30 mM, confirmed in the case of BSA-Hp by multiple techniques. Similar maxima are reported in the literature, but with little discussion, for BSA-poly(dimethyldiallylammonium chloride), lysozyme hyaluronic acid, and lysozyme-chondroitin sulfate, always in the I range 5-30 mM. While inversion of salt effect has been discussed specifically for the interaction of gelatin and sodium polystyrenesulfonate with gelatin(28) and with beta-lactoglobulin,(10) the general nature of this phenomenon, regardless of polyelectrolyte origin, molecular weight, and charge sign has not been recognized. The position of the maxima and their occurrence when protein and polyelectrolyte have the same net charge imply that they arise when Debye lengths extend, at low I, beyond half the protein diameter so that addition of salt screens repulsions, as well as attractions. This appears to be a general effect caused by electrostatic repulsions that can coexist simultaneously with hydrophobic interactions. Modeling of protein electrostatics via Delphi is used to visualize this effect for BSA, lysozyme, insulin, and beta-lactoglobulin. PMID- 12625723 TI - Tailoring the degradation of hydrogels formed from multivinyl poly(ethylene glycol) and poly(vinyl alcohol) macromers for cartilage tissue engineering. AB - Tuning the degradation profiles of polymer cell carriers to match cell and tissue growth is an important design parameter for (cartilage) tissue engineering. In this study, degradable hydrogels were fabricated from divinyl, tetrafunctional poly(ethylene glycol) (PEG) and multivinyl, multifunctional poly(vinyl alcohol) (PVA) macromers to form homopolymer and copolymer gels. These gels were characterized by their volumetric swelling ratio and mass loss profiles as a function of degradation time. By variation of the macromer chemistry and functionality, the degradation time changed from less than 1 day for homopolymer PVA gels to 34 days for pure PEG gels. Furthermore, the degrading medium influenced mass loss, and a marked decrease in degradation time, from 34 to 12 days, was observed with the PEG gels when a chondrocyte-specific medium containing fetal bovine serum was employed. Interestingly, when copolymer gels of PEG and PVA were formed, PVA was released throughout the degradation (as determined by gel permeation chromatography) suggesting that covalent cross linking of the PVA in the network was facilitated by copolymerizing with the PEG macromer. To assess these novel gels for cartilage tissue engineering applications, chondrocytes were photoencapsulated in the copolymer networks and cultured in vitro for up to 6 weeks. DNA, glycosaminoglycan (GAG), and total collagen contents increased with culture time, and the resulting neocartilaginous tissue at 6 weeks was homogeneously distributed as seen histologically. Biochemical analysis revealed that the constructs were comprised of 0.66 +/- 0.04 microg of DNA/mg wet weight (ww), 1.0 +/- 0.05% GAG/ww, and 0.29 +/- 0.07% total collagen/ww at 6 weeks. Furthermore, the compressive modulus increased during culture from 7 to 97 kPa as the neocartilaginous tissue evolved and the gel degraded. In summary, fabricating hydrogels through the copolymerization of PEG and PVA macromers is an effective tool for encapsulating chondrocytes, controlling gel degradation profiles, and generating cartilaginous tissue. PMID- 12625724 TI - Complex coacervation of whey proteins and gum arabic. AB - Mixtures of gum arabic and whey protein (whey protein isolate, WP) form an electrostatic complex in a specific pH range. Three phase boundaries (pH(c), pHphi(1), pHphi(2)) have been determined using an original titration method, newly applied to complex coacervation. It consists of monitoring the turbidity and light scattering intensity under slow acidification in situ with glucono delta-lactone. Furthermore, the particle size could also be measured in parallel by dynamic light scattering. When the pH is lowered, whey proteins and gum arabic first form soluble complexes. This boundary is designated as pH(c). When the interaction is stronger (at lower pH), phase separation takes place (at pHphi(1)). Finally, at pHphi(2) complexation was suppressed by the charge reduction of the gum arabic. The major constituent of the whey protein preparation used was beta-lactoglobulin (beta-lg), and it was shown that beta-lg was indeed the main complex-forming protein. Moreover, an increase of the ionic strength shifted the pH boundaries to lower pH values, which was summarized in a state diagram. The experimental pH(c) values were compared to a newly developed theory for polyelectrolyte adsorption on heterogeneous surfaces. Finally, the influence of the total biopolymer concentration (0-20% w/w) was represented in a phase diagram. For concentrations below 12%, the results are consistent with the theory on complex coacervation developed by Overbeek and Voorn. However, for concentrations above 12%, phase diagrams surprisingly revealed a "metastable" region delimited by a percolation line. Overall, a strong similarity is seen between the behavior of this system and a colloidal gas-liquid phase separation. PMID- 12625725 TI - Kinetics of adsorption, desorption, and exchange of alpha-chymotrypsin and lysozyme on poly(ethyleneterephthalate) tracked film and track-etched membrane. AB - Adsorption kinetics of (125)I-radiolabeled alpha-chymotrypsin at pH 8.6 was studied in a laminar regime between two walls of poly(ethyleneterephthalate) tracked films and membranes. Adsorption kinetics in the presence of solution (10 microg/mL), desorption by rinsing with buffer, and the following exchange of proteins by flowing unlabeled solution were measured. At pH 8.6, alpha chymotrypsin is almost neutral and can be mostly removed from the film surface, contrary to positive lysozyme adsorbed at pH 7.4. Results suggest that alpha chymotrypsin is irreversibly adsorbed in pores, while desorption and exchange occur on membrane flat faces. A method is proposed to determine adsorption kinetics in the pores. Kinetics of desorption and exchange of alpha-chymotrypsin from the film surface can be described by stretched exponential functions in the examined time domain with the same exponent, beta approximately 0.62, which does not depend also on the former adsorption duration. However, the mean residence time at the interface is about 2.5 times greater in the presence of only the buffer than that in the presence of solution. This effect could be explained by a fast exchange at the arrival of unlabeled solution for a part of the adsorbed population. PMID- 12625726 TI - Oxidative degradation of cis- and trans-1,4-polyisoprenes and vulcanized natural rubber with enzyme-mediator systems. AB - Oxidative degradation of cis- and trans-1,4-polyisoprenes by two types of enzyme mediator systems, lipoxygenase/linoleic acid and horseradish peroxidase/1 hydroxybenzotriazole, was investigated at 37 degrees C in aqueous media and analyzed by gel permeation chromatography. Lipoxygenase and horseradish peroxidase activate their substrates, linoleic acid and 1-hydroxybenzotriazole, respectively, for scission of main chains of both 1,4-polyisoprenes. Molecular weights of 1,4-polyisoprenes decreased during the treatment under both enzyme mediator systems, and the depolymerization was completely inhibited by the addition of butylated hydroxytoluene. When the enzyme or the mediator from a reaction system was omitted, depolymerization did not progress, indicating that the scission of polymer chain is induced by the radicals generated only in the presence of both enzyme and mediator. Fenton reagent with linoleic acid was also effective against the degradation of both 1,4-polyisoprenes. Vulcanized natural rubber latex gloves were treated under these three methods, and surface degradation with hole formation was observed with a scanning electron micrograph. PMID- 12625727 TI - Degradation of vulcanized and nonvulcanized polyisoprene rubbers by lipid peroxidation catalyzed by oxidative enzymes and transition metals. AB - Despite numerous reports concerning the biodegradation of rubber materials, there has been no report of rubber degradation by fully characterized enzymes. In the present paper, we presented a new method to decompose nonvulcanized and vulcanized polyisoprene rubbers by controlling the free radical chain reactions of lipids using oxidative enzymes, manganese peroxidase (MnP), laccase (Lac), and horseradish peroxidase (HRP). Nonvulcanized synthetic polyisoprene (IR) was degraded by the free radicals from unsaturated fatty acids produced by MnP, HRP, and a combination of Lac/1-hydroxybenzotriazole. In contrast, lipoxygenase caused no apparent degradation. Degradation of IR was also observed in lipid peroxidation initiated by the Fenton reaction (FR) and Mn(III), an oxidation product produced by MnP. Vulcanized polyisoprene rubber sheets were degraded by the lipid peroxidation initiated by HRP, MnP, Mn(III), and FR. Pyrolysis GC-MS analysis demonstrated that the lipid peroxidation liberated isoprenoid fragments from the vulcanized rubbers. PMID- 12625728 TI - A smart bioconjugate of chymotrypsin. AB - alpha-Chymotrypsin was immobilized on Eudragit S-100 via covalent coupling with 93% retention of proteolytic activity. The conjugate behaved as a smart biocatalyst and functioned as a pH-dependent reversibly soluble-insoluble biocatalyst. The pH optimum of chymotrypsin broadened on immobilization, and the immobilized preparation showed better stability at and above pH 6.5 as compared to the free enzyme. The immobilized enzyme showed a slight shift in the temperature optimum and enhanced thermal stability retaining 70% of its original activity after 1 h of exposure to 40 degrees C as compared to the 25% residual activity for the free enzyme under identical conditions. K(m) and V(max) values did not change on immobilization. Also, the immobilized preparation was quite stable to reuse, it retained almost 85% of its original activity even after a fifth precipitation cycle. UV spectroscopy and circular dichroism were used to probe structural changes in the enzyme upon immobilization. PMID- 12625729 TI - Thermoreversible gelation of aqueous mixtures of pectin and chitosan. Rheology. AB - The synergistic interaction between pectin and chitosan in aqueous acid solution and in the gel phase has been studied by oscillatory shear measurements. Mixtures of pectin and chitosan form thermoreversible gels over a broad composition range by lowering the temperature. The value of the gelation temperature depends on the composition of the mixture, with low values for mixtures with low pectin contents. For incipient gels, a power law can describe the frequency dependence of the complex viscosity, with power law exponents close to -1. The gel evolution of pectin-chitosan mixtures upon a temperature quench below the gel point has been studied. Evidence is provided for a relation between gelation and phase separation in the process of temperature-induced gelation of pectin-chitosan mixtures. A simple model is proposed to rationalize the gelation process in these systems. PMID- 12625730 TI - Copolymerization of 2-carboxyisopropylacrylamide with N-isopropylacrylamide accelerates cell detachment from grafted surfaces by reducing temperature. AB - Acrylic acid (AAc) has been utilized to introduce reactive carboxyl groups to a temperature-responsive polymer, poly(N-isopropylacrylamide) (PIPAAm). However, AAc introduction shifts the copolymer phase transition temperatures higher and dampens the steep homopolymer phase transition with increasing AAc content. We previously synthesized 2-carboxyisopropylacrylamide (CIPAAm) having both a similar side chain structure to IPAAm and a functional carboxylate group in order to overcome these shortcomings. In the present study, these copolymers, grafted onto cell culture plastic, were assessed for cell adhesion control using their phase transition. AAc introduction to PIPAAm-grafted surfaces resulted in excessive surface hydration and hindered cell spreading in culture at 37 degrees C. In contrast, CIPAAm-containing copolymer-grafted surfaces exhibited relatively weak hydrophobicity similar to both homopolymer PIPAAm-grafted surfaces as well as commercial ungrafted tissue culture polystyrene dish surfaces. Cells adhered and spread well on these surfaces at 37 degrees C in culture. As observed previously on PIPAAm-grafted surfaces, cells were spontaneously detached from the copolymer-grafted surfaces by reducing culture temperature. Cell detachment was accelerated on the CIPAAm copolymer-grafted surfaces compared to pure IPAAm surfaces, suggesting that hydrophilic carboxyl group microenvironment in the monomer and polymer is important to accelerate grafted surface hydration below the lower critical solution temperature, detaching cells. PMID- 12625731 TI - Dynamic light scattering of native silk fibroin solution extracted from different parts of the middle division of the silk gland of the Bombyx mori silkworm. AB - Dynamic light scattering (DLS) measurements were performed on aqueous solutions of native silk fibroin extracted from three parts, the posterior (MP), the middle (MM), and the anterior parts (MA), of the middle division (M) of the silk gland of the Bombyx mori silkworm to study the dynamics and aggregation properties of silk fibroin. In the MP part, fibroin molecules are present as aggregates (or clusters) being composed of several large protein complexes or elementary unit (EU), which are further associated to make a large assembly connected via divalent metallic ions. In the MM part, such clusters of EU take more compact structure, and finally in the MA part, clusters disappear, but EUs are more or less aligned to keep the assembly, and the EU takes the conformation of wormlike cylinder capped with hemispheres at both ends. The overall conformational change in solution structure was interpreted as being due to the change in ionic environment in the solution. DLS study was also performed on regenerated silk fibroin solutions, which revealed that fibroin is present as a single molecule dominantly and their association behavior seems completely different from that of native samples and does not depend on types and concentration of added metallic ions. PMID- 12625733 TI - Azobenzene-modified poly(l-glutamic acid) (AZOPLGA): its conformational and photodynamic properties. AB - Azobenzene-modified poly(l-glutamic acid) (AZOPLGA) polymers with 22 and 35 mol % of azo chromophores in the side chains have been synthesized by condensing 4 methoxy-4'-aminoazobenzene and poly(l-glutamic acid). These polymers have been characterized by NMR, FT-IR, and UV-visible spectroscopic techniques. The conformational features of the polymer backbone chains in the films that were cast from the polymer solutions prepared in different solvents have been investigated by circular dichroism spectroscopy. Experimental data suggested that the thermal cis-trans relaxation and photoinduced birefringence, which are related to the azo chromophores in the side chains of polymer, are not affected by the conformations of polymer backbones. However, the modulations of the surface relief gratings, the result of photoinduced mass transport process, recorded on these polymers are sensitive to polymer main chain conformation, as well as the degree of functionalization. PMID- 12625732 TI - Self-organization of amphiphilic polymer in vesicle bilayers composed of surfactant mixtures. AB - Cryogenic transmission electron microscopy (cryo-TEM) and small angle neutron scattering (SANS) are used to investigate the association of amphiphilic polymers consisting of a double-chain hydrophobic tail attached onto poly(ethylene glycol) (PEG) polymer chains into two different systems of equilibrium vesicles. For cetyltrimethylammonium bromide (CTAB)/sodium perfluorohexanoate (FC(5)) vesicle bilayers, the size distribution of the vesicles slightly becomes narrow in the presence of the polymers, suggesting that the wedge-shaped polymers increase the spontaneous curvature of the vesicles. In contrast, the confinement of polymer molecules inside the CTAB/sodium perfluorooctanoate (FC(7)) vesicles that are stabilized by spontaneous curvature causes an abrupt decrease in the bilayer rigidity. By an analysis of vesicle size distribution, it is found that the membrane elasticity of CTAB/FC(7) vesicles is varied considerably from 6k(B)T to 0.3k(B)T, implying the transition of stabilization mechanism from spontaneous curvature to thermal fluctuation in the presence of polymer. The polymer incorporation mechanism into the bilayers is understood, in the comparison of the vesicle radius and size distribution before and after adding polymer, as that the polymer is anchored into the vesicle bilayer owing to hydrophobic property after the adsorption on the surface of the bilayer. PMID- 12625734 TI - Lipase-catalyzed biodegradation of poly(epsilon-caprolactone) blended with various polylactide-based polymers. AB - Poly(epsilon-caprolactone) was blended with various polylactide-based polymers and processed to films by the solution casting method. Blends of 25/75, 50/50, 75/25, 90/10, and 95/5 (w/w) poly(epsilon-caprolactone)/poly(l-lactide), a 95/5 (w/w) blend of poly(epsilon-caprolactone) with a poly(d-lactide), a 50/50 (w/w) poly(l-lactide)-poly(d-lactide) mixture, and a poly(l-lactide-co-epsilon caprolactone) copolymer were considered comparatively. The various phase separated films were allowed to degrade in the presence of Pseudomonas lipase, biodegradation being monitored by proton nuclear magnetic resonance, size exclusion chromatography, differential scanning calorimetry, X-ray diffraction, and environmental scanning electron microscopy. The formation of separated phases during solvent evaporation and their morphologies are discussed. The introduction of poly(l-lactide) dramatically decreased the degradation rate of poly(epsilon caprolactone)/poly(l-lactide) blends. The higher the percentage of poly(l lactide), the slower the degradation, while the presence of cracks and increasing the lipase concentration acted in favor of the enzymatic degradation. Long-term enzymatic degradation of the various 95/5 blends was investigated over 480 h. The poly(epsilon-caprolactone) phase was enzymatically degraded by the lipase regardless of the blend type, the degradation rate depending on the nature of the co-components. PMID- 12625735 TI - Surface engineering of poly(DL-lactide) via electrostatic self-assembly of extracellular matrix-like molecules. AB - We report the development of new biomacromolecule coatings on biodegradable biomaterials based on electrostatic assembly of extracellular matrix-like molecules. Poly(ethylene imine) (PEI) was employed to engineer poly(dl-lactide) (PDL-LA) substrate to obtain a stable positively charged surface. An extracellular matrix- (ECM-) like biomacromolecule, gelatin, was selected as the polyelectrolyte to deposit on the activated PDL-LA substrate via the electrostatic assemble technique. The extracellular matrix-like multilayer on the PDL-LA substrate was investigated by attenuated total reflection (ATR-FTIR), X ray photoelectron spectrscopy (XPS), contact angle, and atomic force microscopy (AFM). The gradual buildup of the protein layer was investigated by UV-vis spectra, and it was further given a quantitative analysis of the protein layer on the PDL-LA substrate via the radioiodination technique. The stability of the protein layer under aqueous condition was also tested by the radiolabeling method. Chondrocyte was selected as the model system for testing the cell behavior and morphology on modified PDL-LA substrates. The chondrocyte test about cell attachment, proliferation, cell activity and cell morphology by SEM, and confocal laser scanning microscopy (CLSM) investigation on extracellular matrix like multilayer modified PDL-LA substrate was shown to promote chondrocyte attachment and growth. Comparing conventional coating methods, polyelectrolyte multiplayers are easy and stable to prepare. It may be a good choice for the modification of 3-D scaffolds used in tissue engineering. These very flexible systems allow broad medical applications for drug delivery and tissue engineering. PMID- 12625736 TI - Analyis of structure/property relationships in silkworm (Bombyx mori) and spider dragline (Nephila edulis) silks using Raman spectroscopy. AB - The molecular deformation of both silkworm (Bombyx mori) and spider dragline (Nephila edulis) silks has been studied using a combination of mechanical deformation and Raman spectroscopy. The stress/strain curves for both kinds of silk showed elastic behavior followed by plastic deformation. It was found that both materials have well-defined Raman spectra and that some of the bands in the spectra shift to lower frequency under the action of tensile stress or strain. The band shift was linearly dependent upon stress for both types of silk fiber. This observation provides a unique insight into the effect of tensile deformation upon molecular structure and the relationship between structure and mechanical properties. Two similar bands in the Raman spectra of both types of silk in the region of 1000-1300 cm(-1) had significant identical rates of Raman band shift of about 7 cm(-1)/GPa and 14 cm(-1)/GPa demonstrating the similarity between the silk fibers from two different animals. PMID- 12625737 TI - Preparation of comb-type N-isopropylacrylamide hydrogel beads and their application for size-selective separation media. AB - A series of the comb-type poly(N-isopropylacrylamide) (NIPAM) gel beads were prepared by inverse suspension polymerization techniques. The comb-type NIPAM gel beads exhibited large volume change at 30 degrees C, and their deswelling rate, defined as the time required for half-shrinking, was 10 times faster than that of the normal-type NIPAM gel beads. The gel beads were utilized to concentrate dilute aqueous solutions of albumin, gamma-globulin, and vitamin B(12). The separation efficiencies of albumin and gamma -globulin with the comb-type NIPAM gel were 80% and 85%, respectively. Whereas those with normal-type NIPAM gel were 55% and 60%, respectively. The incorporation of grafted chains into gel makes the effective mesh size smaller. Therefore it induces the additional obstruction effects between the solutes and network and excludes the high molecular weight solutes. After they have extracted water, their rapid deswelling property makes the gel regenerate effectively by warming to release the absorbed water. PMID- 12625738 TI - Asymmetrical-flow field-flow fractionation with on-line multiangle light scattering detection. 1. Application to wormlike chain analysis of weakly stiff polymer chains. AB - Four samples of hyaluronan in the sodium form, ranging in weight-average molecular weight, M(w), from 6.67 x 10(5) to 4.23 x 10(6) were investigated by asymmetrical-flow field-flow fractionation coupled to multiangle light scattering (FlFFF-MALS) in 0.2 M aqueous NaCl at 25 degrees C. M(w) and z-average radii of gyration, R(G)(z)(), obtained via FlFFF-MALS showed a good agreement with the results obtained by conventional static light scattering. Furthermore, the molecular weight dependence of the radius of gyration for sodium hyaluronan obtained via FlFFF-MALS was analyzed on the basis of the Kratky-Porod model for unperturbed wormlike chains combined with the Yamakawa theory for radius expansion factor, and a sufficiently good agreement was observed between the theoretical prediction and experimental data. These results show the potential usage of FlFFF-MALS regarding size separation and molecular characterization even for weakly stiff chains. PMID- 12625739 TI - Chemoenzymatically synthesized glycoconjugate polymers. AB - Glycoconjugate polymers with poly(vinyl alcohol) (PVA) backbone were synthesized via a chemoenzymatic method. The sugar alcohols of maltose and lactose were submitted to transesterification in the presence of lipases. The esterification was achieved with high selectivity and yield, and the resulting maltitol and lactitol 6-vinyl sebacates were polymerized by a conventional radical initiator with hydrogen peroxide and ascorbic acid. The glycoconjugate polymers carrying alpha-glucose and beta-galactose as recognition signals showed the biological activity such as lectin recognition abilities and hepatocyte adhension. The biodegradability of these polymers was modest but higher than PVA. PMID- 12625740 TI - Structure and morphology changes during in vitro degradation of electrospun poly(glycolide-co-lactide) nanofiber membrane. AB - Electrospun poly(glycolide-co-lactide) (PLA10GA90, LA/GA ratio 10/90) biodegradable nanofiber membranes possessed very high surface area to volume ratios and were completely noncrystalline with a relatively lowered glass transition temperature. These characteristics led to very different structure, morphology, and property changes during in vitro degradation, which were examined systematically. A shrinkage study showed that the electrospun crystallizable but amorphous PLA10GA90 membranes exhibited a very small shrinkage percentage when compared with the electrospun membranes of noncrystallizable poly(lactide-co glycolide) (PLA75GA25, LA/GA 75/25) and poly(d,l-lactide). Although the weight loss of electrospun PLA10GA90 membranes exhibited a similar degradation behavior as cast thin films, detailed studies showed that the structure and morphology changes in electrospun membranes followed different pathways during the hydrolytic degradation. After 1 day of degradation in buffer solution at 37 degrees C, electrospun PLA10GA90 membranes exhibited a sudden increase in crystallinity and glass transition temperature, due to the fast thermally induced crystallization process. The continuous increase in crystallinity and apparent crystal size, as well as the decrease in long period and lamellae thickness, indicated that the thermally induced crystallization was followed by a chain cleavage induced crystallization process. The mass loss rate was accelerated after 6 days of degradation. The increase in glass transition temperature during this period further confirmed that the degradation of PLA10GA90 nanofibers was initiated from the amorphous region within the lamellar superstructures. A mechanism of structure and morphology changes during in vitro degradation of electrospun PLA10GA90 nanofibers is proposed. PMID- 12625741 TI - First-order kinetics analysis of monomer composition dependent polyhydroxyalkanoic acid degradation in Pseudomonas spp. AB - The intracellular degradation of polyhydroxyalkanoic acid (PHA) in pseudomonads was investigated by first-order kinetics analysis using the initial rate method. One type of PHA was accumulated in five Pseudomonas spp., P. oleovorans, P. aeruginosa, P. fluorescens, P. citronellolis, and P. putida, by growing them on octanoic acid. The monomer compositions of the five PHA were not significantly different from one another: 85-90 mol % 3-hydroxyoctanoic acid (3HO), 7-12 mol % 3-hydorxycaproic acid (3HC), and 3-6 mol % 3-hydroxydecanoic acid (3HD). The first-order degradation rate constants (k(1)) for the octanoate-derived PHA (designated P(3HO)) in the five species were in a similar range between 0.060 and 0.088 h(-1). This may indicate the similar specificities of the five intracellular depolymerases. In addition, the similar k(1) among the different species may correlate with the high degree of amino acid sequence identities (over 85%) among the intracellular PHA depolymerase phaZ genes. Six other chemically different types of PHA were accumulated in P. putida from n-nonanoic acid, n-decanoic acid, 5-phenyvaleric acid, or 11-phenoxyundecanoic acid as a single or a mixed carbon source. The calculated k(1) values were characteristic to each PHA, reflecting their chemical structures. In comparison with P(3HO), an increase in the levels of the two minor monomers 3HC and 3HD as in P(21 mol % 3HC co-56 mol % 3HO-co-23 mol % 3HD) significantly slowed the rate of intracellular degradation. From the comparison of k(1) values, it is suggested that the P. putida intracellular depolymerase is most active against P(3HO). PMID- 12625743 TI - Synthesis and characterization of poly(butylene succinate-co-butylene malate): a new biodegradable copolyester bearing hydroxyl pendant groups. AB - A new biodegradable copolyester, poly(butylene succinate-co-butylene malate) P(BS co-BM), has been preliminarily prepared with optically active centers and lateral hydroxyl functional groups via a four-step synthetic strategy. First, an optically active benzyl-protected dimethyl malate was synthesized from a starting material of (S)-dimethyl malate and purified with good yield. Then, copolyester poly(butylene succinate-co-benzyl-protected butylene malate), P(BS-co-BBM), was prepared through a skilled condensation copolymerization of the benzyl-protected dimethyl malate, dimethyl succinate, and 1,4-butanediol in the presence of titanium tetraisopropoxide as the catalyst. Finally, a Pd/C catalyzed hydrogenation was applied to eliminate the benzyl protection group in a mixed solution of THF and methanol; thus the target copolyester P(BS-co-BM) was attained. On the other hand, physical properties of the synthesized copolyesters were systematically characterized by means of nuclear magnetic resonance spectrometer, Fourier transformed infrared spectrometer, gel permeation chromatography, optical polarimeter, quantitative hydroxyl titration, and thermal analytical instruments. The experimental evidence demonstrated a successful construction of the product P(BS-co-BM) bearing lateral hydroxyl functional groups. It was also revealed that the lower BBM unit content was in the benzyl protected optically active P(BS-co-BBM) copolyester, the higher melting point T(m), crystallinity, the broader molecular distribution, and the lower glass transition temperature T(g) would be detected, and these results can be accounted for the presence of bulky lateral benzyl moieties. In contrast, the deprotected product P(BS-co-55 mol % BM) showed a higher T(m), crystallinity and lower T(g) than its counterpart P(BS-co-55 mol % BBM). Interestingly, a thermal stability as high as that of the linear PBS was observed for P(BS-co-55 mol % BM) while a strong BBM unit content dependence of thermal stability was detected for the benzyl-protected copolyester P(BS-co-BBM)s. Therefore, these results may be beneficial for the new optically active P(BS-co-BM) bearing hydrophilic hydroxyl functional groups as a potential biomaterial. PMID- 12625742 TI - Riboflavin-photosensitized changes in aqueous solutions of alginate. Rheological studies. AB - Interactions between photoexcited riboflavin (RF), promoted by irradiation in the range of 310-800 nm, and alginate have been studied in air equilibrated aqueous solutions with the aid of rheological methods. Light irradiation of RF causes under aerobic conditions fragmentation of alginate and a decrease in the shear viscosity and other rheological parameters of its solutions. The decrease is most pronounced in concentrated polymer solutions. The photochemical degradation of alginate is inhibited in the presence of the quenchers/scavengers d-mannitol, glutathione, potassium iodide, and sodium azide and in excess oxygen. The addition of thiourea to alginate-RF solutions leads to enhanced degradation of the polymer. Significant shear-thinning effects and deviations from the Cox-Merz rule are observed at higher polymer concentrations. PMID- 12625744 TI - Layer-by-layer assembly to modify poly(l-lactic acid) surface toward improving its cytocompatibility to human endothelial cells. AB - A novel technique to introduce free amino groups onto polyester scaffolds via aminolyzing the ester groups with diamine has been developed recently. Positively charged chitosan was then deposited onto the aminolyzed poly(l-lactic acid) (PLLA) membrane surface in a layer-by-layer assembly manner using poly(styrene sulfonate, sodium salt) (PSS) as a negatively charged polyelectrolyte. The layer by-layer deposition process of PSS and chitosan was monitored by UV-vis absorbance spectroscopy, energy transfer by fluorescence spectroscopy, and advancing contact angle measurements. The existed chitosan obviously improved the cytocompatibility of PLLA to human endothelial cells. The cell attachment, activity, and proliferation on the PLLA membranes assembled with three or five bilayers of PSS/chitosan with chitosan as the outermost layer were better than those with one bilayer of PSS/chitosan or the control PLLA. The cells also showed morphology of an elongated shape with abundant cytoplasm, and a confluent cell layer was reached after being cultured for 4 days. Measurement of von Willebrand factor secreted by these endothelial cells (ECs) verified the endothelial function. Hence, better ECs compatible PLLA were produced. PMID- 12625745 TI - Shear-induced mixing and demixing in aqueous methyl hydroxypropyl cellulose solutions. AB - The influence of shear flow on the phase separation of aqueous methyl hydroxypropyl cellulose solutions was investigated by means of rheoturbidity and online rheo small angle light scattering (SALS) experiments. In semidilute solution shear-induced mixing was observed and the cloud curve was shifted to higher temperatures with increasing shear rate. With higher concentrated solutions, however, shear-induced demixing was found. The shear-induced mixing is interpreted as being a disruption of slightly entangled clusters under the influence of the shear energy. The shear demixing appears in line with the observation with other systems. A characteristic butterfly pattern was observed in rheo-SALS. PMID- 12625748 TI - On the incorporation of multi-modality image registration into the radiotherapy treatment planning process. AB - A technique is presented that allows the direct use of physiological image sets in the radiation therapy treatment planning process. When fused to the treatment planning CT, physiological image studies may allow one to define physiological tumor subvolumes consisting of areas of possible chronic hypoxia, areas of high perfusion, areas of high diffusion, and areas containing high choline concentrations. These physiological tumor subvolumes could be selectively boosted to increase local control of malignant brain tumors once one has determined which of these physiological tumor subvolumes predicts for local tumor recurrence after conventional radiotherapy. In this technique a user assisted automatic registration technique is used that is based on an analytical estimate for the transformation matrix needed to register two rigid bodies. The only user input needed is three non-collinear points selected based on landmarks in the primary image and the corresponding three points in the secondary image. Since this registration technique uses two sets of at least three user-defined landmark points each of which has some selection error associated with it, the final registration will have an error that depends only on the selection error associated with the point sets. Since physiological image studies are acquired at the same setting as the T1- w MRI their spatial orientation with respect to the T1- w MRI is known. Therefore, the registration of multiple physiological image studies to the treatment planning CT can be accomplished by first correlating them to the T1- w MRI, and in a second step the T1- w MRI is then registered to the treatment planning CT. The desired registration of the physiological image studies to the treatment planning CT is then accomplished by simply composing the appropriate transformation matrices. PMID- 12625746 TI - Use of small-angle neutron scattering to study tubulin polymers. AB - Small-angle neutron scattering has been used to examine taxol-stabilized microtubules and other tubulin samples in both H(2)O and D(2)O buffers. Measurements were made at pH/pD values between 6.0 and 7.8, and observed scattered intensities, I(Q), have been interpreted in terms of multicomponent models of microtubules and related tubulin polymers. A semiquantitative curve fitting procedure has been used to estimate the relative amounts of the supramolecular components of the samples. At both pH and pD 7.0 and above, the tubulin polymers are seen to be predominantly microtubules. Although in H(2)O buffer the polymer distribution is little changed as the pH varies, when pD is lowered the samples appear to contain an appreciable amount of sheetlike structures and the average microtubule protofilament number increases from ca. 12.5 at pD > or = approximately 7.0 to ca. 14 at pD approximately 6.0. Such structural change indicates that analysis of microtubule solutions based on H(2)O/D(2)O contrast variation must be performed with caution, especially at lower pH/pD. PMID- 12625749 TI - Comparison of volume-rendered and surface-rendered MR colonography. AB - In the United States and Europe, colorectal cancer is the second leading cause of cancer-related deaths. It is well known that colorectal carcinomas may originate from preexisting adenomas. For the visualization of colorectal cancer and other pathologic changes such as polyps, two 3D methods (volume-rendering (VR) and surface-rendering (SR)) in MR colonography were compared in our study. MR colonography was carried out in 17 patients on a 1.5 T MR scanner using a 10 mmolar gadolinium water solution enema. Coronal as well as rotated VR and SR views were compared in order to examine the technical quality (TQ) of the visualization model and grade of confidence (GC) in the pathological findings. Colonoscopic findings revealed 8 colorectal carcinoma, 10 patients with polyps, 4 diverticular disease, and 2 with redundant bowel loops. Based on a total of 248 colonic segments for both visualization methods, volume rendering were significantly superior to surface rendering for both, TQ (p<0.0001) and GC (p<0.0001). Volume rendering and surface rendering were not dependent on individual colon segments (p=0.13 for TQ and p=0.18 for GC) or on image rotation (p=0.06 for TQ and p=0.062 for GC). It is also independent of the type of pathology (p=0.31 for TQ and p=0.42 for GC) and the reviewers (p=0.62 for TQ and p=0.88 for GC). This indicates, that for the purpose of interpreting the technical quality and pathological findings, volume rendering is superior to surface rendering in MR colonography. Volume rendering could be used as an 3D visualization tool, enabling MR colonography examinations to be completed sooner in cases where colon distension is sufficient, and it would also provide an overview of potential mass lesions. PMID- 12625751 TI - CyberKnife radiotherapy for localized prostate cancer: rationale and technical feasibility. AB - There is a clear dose response for localized prostate cancer radiotherapy and there probably is a radiobiological rationale for hypo-fractionation. Combining the two should maximize tumor control and increase the therapeutic ratio. This study examines the rationale and technical feasibility of CyberKnife radiotherapy (a robotic arm-driven linear accelerator) for localized prostate cancer. Its ability to deliver non-coplanar non-isocentric arcs can yield maximally conformal isodoses. It is the only integrated system capable of target position verification and real-time tracking during delivery of conformal stereotactic radiotherapy. Inverse planning with the CyberKnife is used to design a course of radiotherapy for localized prostate cancer. Fiducial markers within the gland are used to verify organ position and track organ motion via an orthogonal pair of electronic x-ray imaging devices and provide real-time feedback correction to the robotic arm during delivery. Conformal isodose curves and dose volume histograms (DVH) are used to compare with an optimized Intensity-Modulated Radiotherapy (IMRT) plan actually delivered to the study patient based upon CT scan-derived organ volumes. The CyberKnife can produce superior DVHs for sparing of rectum and bladder and excellent DVHs for target coverage compared with IMRT, and possesses dose heterogeneities to the same degree as IMRT plans. Because of the significantly longer delivery times required it would be best suited for hypo fractionated regimens. Such dose regimens might allow for biologically equivalent dose escalation without increased normal tissue toxicity. Since the CyberKnife can verify organ position and motion and correct for this in real-time it is the ideal means of achieving such excellent DVHs without a compromise in doses to normal tissues. These capabilities are essential if one contemplates hypo fractionated regimens with large dose-per-fraction sizes (>5Gy to 10Gy) and dose escalation. PMID- 12625750 TI - Intraoperative MR-guided instillation of phosphorus-32 for cystic craniopharyngiomas: case report. AB - Intraoperative magnetic resonance imaging has been applied to a number of neurosurgical disease processes since the late 1990's. The ability to visualize the operative site in near-real time has added a significant degree of safety to the treatment of lesions such as a cystic craniopharyngioma which can be located in regions of the brain where an untoward consequence can result in significant neurological morbidity. Previous surgical techniques, although often successful, did not allow the neurosurgeon to directly visualize whether the goals of surgery had been met or whether there was an inadvertent complication associated with the surgical approach until after the event had occurred. The safe and accurate instillation of radioactive phosphorus into this cystic tumor resulted in clinical improvement and the maintenance of normal pituitary function for this patient. The extreme accuracy and safety of this surgical technique is demonstrated by the imaging examples provided. PMID- 12625752 TI - Applications to cancer research of "lab-on-a-chip" devices based on dielectrophoresis (DEP). AB - The recent development of advanced analytical and bioseparation methodologies based on microarrays and biosensors is one of the strategic objectives of the so called post-genomic. In this field, the development of microfabricated devices could bring new opportunities in several application fields, such as predictive oncology, diagnostics and anti-tumor drug research. The so called "Laboratory-on a-chip technology", involving miniaturisation of analytical procedures, is expected to enable highly complex laboratory testing to move from the central laboratory into non-laboratory settings. The main advantages of Lab-on-a-chip devices are integration of multiple steps of different analytical procedures, large variety of applications, sub-microliter consumption of reagents and samples, and portability. One of the requirement for new generation Lab-on-a-chip devices is the possibility to be independent from additional preparative/analytical instruments. Ideally, Lab-on-a-chip devices should be able to perform with high efficiency and reproducibility both actuating and sensing procedures. In this review, we discuss applications of dielectrophoretic(DEP) based Lab-on-a-chip devices to cancer research. The theory of dielectrophoresis as well as the description of several devices, based on spiral-shaped, parallel and arrayed electrodes are here presented. In addition, in this review we describe manipulation of cancer cells using advanced DEP-based Lab-on-a-chip devices in the absence of fluid flow and with the integration of both actuating and sensing procedures. PMID- 12625753 TI - Evaluation of TNF-alpha/Bax gene therapy and radiation against C6 glioma xenografts. AB - Successful therapy of high-grade tumors of the brain is likely to require a combination of new therapeutic approaches. The major goal of the present study was to construct a plasmid-based bax gene vector (pGL1-Bax) and evaluate its expression in vitro and in vivo using athymic mice with subcutaneously growing C6 glioma. Preliminary experiments of efficacy and safety were also performed using pGL1-Bax alone and in combination with previously constructed pGL1-TNF-alpha, as well as with radiation. pGL1-Bax was expressed by C6 cells and was correlated with apoptosis, indicating that the construct and the bax protein were functional. Although intratumoral injections of pGL1-Bax alone, up to total doses of 450 micro g, did not significantly affect tumor growth, consistently smaller tumors were obtained when pGL1-TNF-alpha plus pGL1-Bax were injected 16-18 hr prior to tumor irradiation. Furthermore, in mice with two tumors, one treated and one untreated, progression of the untreated tumor was delayed in the animals receiving all three modalities. No prohibitive toxicities were noted, based on mouse body weights and in vitro assays of blood and spleen. Significant increases in spleen mass, total leukocyte counts, percentage of granulocytes, spontaneous blastogenesis, and CD71-expressing B cells were primarily associated with tumor presence and not treatment type. Overall, the results are promising and suggest that TNF-alpha/Bax gene therapy may be beneficial against highly malignant tumors of the brain. To our knowledge, this is the first report of bax gene therapy used together with radiation in an in vivo glioma model. PMID- 12625754 TI - Cancer gene therapy. AB - Cancer gene therapy is the transfer of genetic material to the cells of an individual with the goal of eradicating cancer cells, both in the primary tumor and metastases. Cancer gene therapy strategies exploit our expanding knowledge of the genetic basis of cancer, thereby allowing rationally targeted interventions at the molecular level. The successful implementation of cancer gene therapy in the clinic awaits the development of vectors capable of specific and efficient gene delivery to cancer cells. The first clinical applications of cancer gene therapy are likely to be in combination with conventional therapies, such as radiotherapy and immunotherapy. PMID- 12625755 TI - Photodynamic therapy for esophageal cancer: a useful and realistic option. AB - The use of light therapy for tissue destruction is highly attractive for the endoscopic and minimally invasive therapy of esophageal cancer. Photodynamic therapy (PDT) offers the possibility of palliation of advanced obstructing tumors. However, there are other competing techniques, which can be used to open the esophageal lumen. It has also proved very effective in providing prolonged palliation of patients with advanced irresectable cancer. Completely obstructing tumors, tortuous and long lesions, and tumors near the upper end of the esophagus are particularly suitable for photodynamic therapy. Patients with obstruction to an esophageal prosthesis are also well palliated with PDT. A more interesting and exciting development is its use for the eradication of early asymptomatic mucosal disease. Photodynamic therapy is particularly useful for the eradication of field cancerous change in patients with pre-malignant Barrett's esophagus, or early tumors in patients unfit for radical therapy. PMID- 12625756 TI - Overview of electroporation. AB - This special issue of Technology in Cancer Research and Treatment focuses on the versatility of electroporation. Contributed papers report on the basic mechanisms as well as the use of this procedure ex vivo and in vivo. In addition, recent studies utilizing in vivo electroporation for plasmid DNA and drug delivery are presented. PMID- 12625757 TI - Factors controlling electropermeabilisation of cell membranes. AB - Electric field pulses are a new approach for drug and gene delivery for cancer therapy. They induce a localized structural alteration of cell membranes. The associated physical mechanisms are well explained and can be safely controlled. A position dependent modulation of the membrane potential difference is induced when an electric field is applied to a cell. Electric field pulses with an overcritical intensity evoke a local membrane alteration. A free exchange of hydrophilic low molecular weight molecules takes place across the membrane. A leakage of cytosolic metabolites and a loading of polar drugs into the cytoplasm are obtained. The fraction of the cell surface which is competent for exchange is a function of the field intensity. The level of local exchange is strongly controlled by the pulse duration and the number of successive pulses. The permeabilised state is long lived. Its lifetime is under the control of the cumulated pulse duration. Cell viability can be preserved. Gene transfer is obtained but its mechanism is not a free diffusion. Plasmids are electrophoretically accumulated against the permeabilised cell surface and form aggregates due to the field effect. After the pulses, several steps follow: translocation to the cytoplasm, traffic to the nucleus and expression. Molecular structural and metabolic changes in cells remain mostly poorly understood. Nevertheless, while most studies were established on cells in culture (in vitro), recent experiments show that similar effects are obtained on tissue (in vivo). Transfer remains controlled by the physical parameters of the electrical treatment. PMID- 12625758 TI - Digression on membrane electroporation for drug and gene delivery. AB - Membrane electroporation (ME) defines an electrical technique to render lipid membranes porous and permeable, transiently and reversibly, by external voltage pulses. Although there are numerous applications of ME to manipulate cells, organelles and tissues in cell biology, biotechnology and medicine, yet the molecular mechanism of ME is only slowly being understood. A general chemical- thermodynamical approach for the quantitative description of cell membrane electroporation has been developed to provide the framework to quantitatively rationalize electroporative cell transformation and electroporative uptake of drug-like dyes into cells, as well as electrolyte efflux from salt-filled electroporated vesicles. Mechanistically, the electroporative transfer of gene and drug-like dyes involves the coupling between an interactive contact formation of the permeates with the cell surface membrane and the structural electroporation-resealing cycle C <--> (P) where C is the closed and (P) represents a number of different porated membrane states, respectively. The experimentally accessible concentration fraction f(p) = [(P)] / ([C] + [(P)]) of porous states is related to thermodynamic and electro-mechanic parameters such as temperature and the electric field strength, membrane rigidity or curvature. The results of the theoretical approach, mainly based on electrooptical data of lipid vesicles, have been successfully used to analyze single cells and to specify conditions for the practical purpose of direct electroporative gene transfer and drug delivery, in particular in the new medical disciplines of electroporative chemotherapy and electroporative gene vaccination. PMID- 12625759 TI - Highly efficient, large volume flow electroporation. AB - Electroporation is widely used to transfect and load cells with various molecules. Traditional electroporation using a static mode is typically restricted to volumes less than 1 mL, which limits its use in clinical and industrial bioprocessing applications. Here we report efficient, large volume transfection results by using a scalable-volume electroporation system. Suspended (Jurkat) and adherent cells (10T1/2 and Huh-7) were tested. A large macromolecule, FITC-conjugated dextran (MW=500 kD) was used to measure cell uptake, while a plasmid carrying the gene coding for enhanced green fluorescence protein (eGFP) was used to quantitate the flow electrotransfection efficiency as determined by flow cytometry. The flow electroloading efficiency of FITC-dextran was >90%, while the cell viability was highly maintained (>90%). High flow electrotransfection efficiency (up to 75%) and cell viability (up to 90%) were obtained with processing volumes ranging from 1.5 to 50 mL. No significant difference of electrotransfection efficiency was observed between flow and static electrotransfection. When 50 mL of cell volume was processed and samples collected at different time points during electroporation, the transgene expression and cell viability results were identical. We also demonstrated that DNA plasmid containing EBNA1-OriP elements from Epstein-Barr virus were more efficient in transgene expression than standard plasmid without the elements (at least 500 too 1000-fold increase in expression level). Finally, to examine the feasibility of utilizing flow electrotransfected cells as a gene delivery vehicle, 10T1/2 cells were transfected with a DNA plasmid containing the gene coding for mIL12. mIL12 transfected cells were injected subcutaneously into mice, and produced functional mIL12, as demonstrated by anti-angiogenic activity. This is the first demonstration of efficient, large volume, flow electroporation and the in vivo efficacy of flow electrotransfected cells. This technology may be useful for clinical gene therapy and large-scale bioprocesses. PMID- 12625760 TI - Applications of plasmid electrotransfer. AB - The use of electric pulses to transfect cells has recently been extended to show the utility of this procedure in vivo. Electrotransfer has been performed in vivo on several tissue types including skin, blood vessels, liver, tumor, muscle, cornea, brain and spleen. The most widely targeted tissue has been skeletal muscle. In addition to its potential use in gene therapy, in vivo DNA electrotransfer is also, because of its simplicity, a powerful laboratory tool to study in vivo gene expression and function in a given tissue. Many published studies have now shown that plasmid electrotransfer can lead to a long-lasting therapeutic effect in various pathologies, such as cancer, blood disease, or muscle ischemia. The future potential for this gene therapy approach will include delivery for both local action or distal effect by secretion of the transgenic proteins in the circulation. PMID- 12625761 TI - Electric field enhanced plasmid delivery to liver hepatocellular carcinomas. AB - Electric field enhanced molecular delivery for cancer research and treatment is a new technology that has demonstrated its effectiveness in clinical trials using bleomycin or cisplatin (Heller, R., Gilbert, R., Jaroszeski, M. J. Clinical applications of electrochemotherapy, Advanced Drug Delivery Reviews 35,119-129, 1999), as chemotherapeutic agents. The technology is being investigated in research applications for applicability as a method to enhance gene expression in a target tumor. Success is predicated on an appropriate effective electric field mediated delivery protocol that triggers significant appropriate gene expression duration and levels. An electric field mediated delivery protocol includes a set of conditions associated with the electric field, the electroporation signature, as well as parameters associated with the plasmid and the electric field applicator. Manipulation of the electrical parameters within the electroporation signature generates different gene expression levels in liver hepatocellular carcinomas. Statistically significant gene expression levels were obtained that differed by an order of magnitude when two different electric field strength and duration conditions were employed. PMID- 12625762 TI - Electrical enhancement of formulated plasmid delivery in animals. AB - Electroporation has been shown to significantly increase plasmid transfer to the skeletal muscle, but this procedure is also implicated in muscle damage. We are reporting a highly efficient in vivo transfer of a plasmid formulated with poly (L-glutamate) (PLG) into murine, canine and porcine muscle fibers using electric pulses of low field intensity. In mice and pigs, the use of secreted embryonic alkaline phosphatase (SEAP) as the indicator gene caused increased PLG expression by 2-3 fold compared to naked plasmid; while delivery of a PLG-plasmid formulation to dogs showed a 10-fold increase in serum SEAP levels compared to plasmid alone. Muscle lesions were reduced by the protective PLG. Thus, PLG may constitute a useful adjuvant for increased expression and reduced muscle trauma to plasmid DNA delivered by electroporation. PMID- 12625764 TI - Effect of electrochemotherapy on muscle and skin. AB - The efficient delivery of drugs to tumors is an important tool for the treatment of a variety of cancers. Electric pulses have been shown to facilitate the uptake of molecules through the cell membrane. This procedure has been successful in increasing the effectiveness of anti-tumor agents (electrochemotherapy; ECT). Response rates of >80% have been obtained in both animal and human trials for several types of skin malignancies. The study reported here examined the effect of ECT on normal tissue. The hind limbs of Sprague Dawley rats were treated with 1-3 electroporation sequences in the presence or absence of the drug (bleomycin) which was administered at 4, 8 or 16 units/ml. The treated sites were examined histologically 3, 14 and 56 days later. Limb function was not affected by the treatment and skin and muscle necrosis was only seen at the higher doses. PMID- 12625763 TI - The application of electroporation to transfect hematopoietic cells and to deliver drugs and vaccines transcutaneously for cancer treatment. AB - Electroporation and the associated phenomenon of electrofusion have been widely adapted as tools to a broad range of biomedical research and therapy. In this article, we summarize our adaptation of the electroporation and electrofusion technology in two fronts of cancer research and treatment. The first is genetic manipulation of hematopoietic cells for the purpose of cancer treatment. High efficiency transfection methods have been developed to transfect NK cells, peripheral blood stem cells, and bone marrow derived dendritic cells. Hybrids of tumor cells and bone marrow derived dendritic cells have been formed by electrofusion for the purpose of tumor vaccines. The second front is the use of transcutaneous electroporation to deliver anticancer drugs and vaccines across the skin. Methods to extend the upper molecular weight limit of transcutaneous electroporation have been developed. The pro-photosensitizer drug, delta-amino levulinic acid, the anticancer drug methotrexate, and peptide vaccines designed for cancer prevention and immunotherapy have been delivered transcutaneously by electroporation. These studies hold promise for the treatment of cancers in human. PMID- 12625766 TI - Modification of reference array attachment for image guided neurosurgery: direct cranial fixation. AB - We have modified the cranial fixation technique of the reference array used for the Stealth (Medtronics Inc., Minneapolis, MN) image guided neurosurgical workstation to avoid rigid immobilization and to accommodate patients undergoing awake procedures. The modification allows attachment of a reference array directly to the skull prior to registration, avoiding the requirement for rigid cranial fixation. The accuracy of fiducial registration for the modified reference array was compared to the conventional reference array using a phantom system yielding similar registration results and target accuracy. The successful application of the modified system to two operative cases is described. PMID- 12625765 TI - Electrochemotherapy for the treatment of human sarcoma in athymic rats. AB - Electrochemotherapy is the combined use of a chemotherapeutic agent and pulsed electric fields. Electrical treatment causes an increase in cell membrane permeability which allows the chemotherapeutic agent to more freely enter the tumor cells. Electrochemotherapy has been under development in clinical trials. This study focused on determining the applicability of electrochemotherapy for treating soft tissue sarcoma using an animal model bearing human sarcomas. The antitumor effects of several concentrations of cisplatin, bleomycin, doxorubicin, and netropsin as single agents delivered with electric pulses were investigated based on post-treatment tumor volumes and histology. Electrochemotherapy treatment resulted in 5% to 88.9% durable complete responses; ECT that employed bleomycin resulted in the highest antitumor effects. This indicates the feasibility of electrochemotherapy as a modality for limb preserving treatments for sarcoma of the extremities. PMID- 12625767 TI - Transcription factor decoy (TFD) in breast cancer research and treatment. AB - Synthetic oligonucleotides have recently been the object of many investigations aimed to develop sequence-selective compounds able to modulate, either positively or negatively, transcription of eukaryotic and viral genes. Alteration of transcription could be obtained by using synthetic oligonucleotides mimicking target sites of transcription factors (the transcription factor decoy -TFD- approach). This could lead to either inhibition or activation of gene expression, depending on the biological functions of the target transcription factors. Since several transcription factors are involved in tumor onset and progression, this issue is of great interest in order to design anti-tumor compounds. In addition to oligonucleotides, peptide nucleic acids (PNA) can be proposed for the modulation of gene expression. In this respect, double-stranded PNA-DNA chimeras have been shown to be capable to exhibit strong decoy activity. In the case of treatment of breast cancer cells, decoy oligonucleotides mimicking CRE binding sites, promoter region of estrogen receptor alpha gene, NF-kB binding sites have been used with promising results. Therefore, the transcription factor decoy approach could be object of further studies to develop protocols for the treatment of breast cancer. In the future, transcription factors regulating cell cycle, hormone-dependent differentiation, tumor invasion and metastasis are expected to be suitable targets for transcription factor decoy. PMID- 12625769 TI - Challenges and opportunities for in vivo imaging in oncology. AB - Advances in genomics, proteomics and technology are changing medicine in fundamental ways. There are increasing clinical and laboratory requirements to obtain cellular and molecular information in vivo. This is particularly true in oncology, where the behavior of tumor cells is inextricably linked to their milieu. If cancer cells are removed from their microenvironment, their pattern of gene expression changes. Therefore, non-invasive, quantitative means of detecting gene and protein activity are essential. In vivo imaging is one methodology for achieving this. Marked advances in tracer methods for PET scanning or single photon nuclear medicine techniques have occurred in the past few years. MRI contrast agents that reflect physiologic information are also being developed, although larger mass quantities of injectable material are required. The useful concept of "activatable agents" was pioneered in MRI. Similarly, ultrasound and computed tomography are being re-engineered to reflect information at the cellular level. In vivo optical imaging technologies have matured to the point where they are indispensable laboratory tools for small animal imaging. Human applications are in the feasibility testing stage, and the future for clinical optical imaging techniques looks bright. Merging these molecular imaging techniques with minimally or non-invasive image-guided therapeutic delivery techniques is a subsequent goal in the fight against cancer. PMID- 12625770 TI - Positron emission tomography in cancer research and treatment. AB - Positron emission tomography (PET), the imaging of pharmaceuticals labeled with positron-emitting radionuclides, is a rapidly growing modality for the diagnosis and management of cancer. PET yields high-quality images characterizing substrate metabolism, cellular proliferation, receptor density, and other parameters that can be used to identify cancer and evaluate its response to therapies. The technique mainly utilized in cancer management is FDG-PET, which exploits the abnormal glucose metabolism of cancer cells first characterized by Warburg. We discuss the principles of PET, the currently available instrumentation and radiopharmaceuticals, the efficacy of FDG-PET in the management of cancer, and the prospects for near-term advances in cancer using PET. PMID- 12625771 TI - Development and Monte Carlo analysis of antiscatter grids for mammography. AB - Mammography arguably demands the highest fidelity of all x-ray imaging applications, with simultaneous requirements of exceedingly high spatial and contrast resolution. Continuing technical improvements of screen-film and digital mammography systems have led to substantial improvements in image quality, and therefore improvements in the performance of anti-scatter grids are required to keep pace with the improvements in other components of the imaging chain. The development of an air-core honeycomb (cellular) grid using x-ray lithography and electroforming techniques is described, and the production of a 60 mm x 60 mm section of grid is reported. A crossed grid was constructed with 25 microm copper septa, and a period of 550 microm. Monte Carlo and numerical simulation methods were used to analyze the theoretical performance of the fabricated grid, and comparisons with other grid systems (Lorad HTC and carbon fiber interspaced grids) were made over a range of grid ratios. The results demonstrate essentially equivalent performance in terms of contrast improvement factor (CIF) and Bucky factor (BF) between Cu and Au honeycomb grids and the Lorad HTC (itself a copper honeycomb grid). Gold septa improved both CIF and BF performance in higher kVp, higher scatter geometries. The selectivity of honeycomb grids was far better than for linear grids, with a factor of approximately 3.9 improvement at a grid ratio of 5.0. It is concluded that using the fabrication methods described, that practical honeycomb grid structures could be produced for use in mammographic imaging, and that a substantial improvement in scatter rejection would be achieved using these devices. PMID- 12625772 TI - Dual-modality imaging of cancer with SPECT/CT. AB - Dual-modality imaging is an in vivo diagnostic technique that obtains structural and functional information directly from patient studies in a way that cannot be achieved with separate imaging systems alone. Dual-modality imaging systems are configured by combining computed tomography (CT) with radionuclide imaging (using positron emission tomography (PET) or single-photon emission computed tomography (SPECT)) on a single gantry which allows both functional and structural imaging to be performed during a single imaging session without having the patient leave the imaging system. A SPECT/CT system developed at UCSF is being used in a study to determine if dual-modality imaging offers advantages for assessment of patients with prostate cancer using (111)In-ProstaScint, a radiolabeled antibody for the prostate-specific membrane antigen. (111)In-ProstaScint images are reconstructed using an iterative maximum-likelihood expectation-maximization (ML EM) algorithm with correction for photon attenuation using a patient-specific map of attenuation coefficients derived from CT. The ML-EM algorithm accounts for the dual-photon nature of the 111In-labeled radionuclide, and incorporates correction for the geometric response of the radionuclide collimator. The radionuclide image then can be coregistered and overlaid in color on a grayscale CT image for improved localization of the functional information from SPECT. Radionuclide images obtained with SPECT/CT and reconstructed using ML-EM with correction for photon attenuation and collimator response improve image quality in comparison to conventional radionuclide images obtained with filtered backprojection reconstruction. These results illustrate the potential advantages of dual modality imaging for improving the quality and the localization of radionuclide uptake for staging disease, planning treatment, and monitoring therapeutic response in patients with cancer. PMID- 12625773 TI - Contrast-assisted destruction-replenishment ultrasound for the assessment of tumor microvasculature in a rat model. AB - Angiogenesis, the development of new blood vessels, is necessary for tumor growth. Anti-angiogenic therapies have recently received attention as a possible cancer treatment. The purpose of this study was to monitor the vascularity of induced tumors in rats using contrast-enhanced ultrasound during anti-angiogenic therapy. Six rats with subcutaneously implanted R3230 murine mammary adenocarcinomas were treated with an orally administered anti-angiogenic agent (SU11657) beginning 28 days after tumor implantation (20 mg/kg BW once daily). Three additional tumor-bearing control rats were treated with an equivalent volume of vehicle alone. Sonographic evaluation of tumor blood flow was performed using a modified Siemens Sonoline Elegra equipped with a 5.0 MHz linear transducer prior to drug administration, during the first 51 hours following initial drug administration, and on days 8 and 15 after initiation of therapy. Tumor volumes were estimated at each time point using a prolate ellipsoid method from linear dimensions measured on the B-mode ultrasound image in the three major axes. A destruction-replenishment technique was used for tumor blood flow evaluation using a constant rate infusion of intravenously delivered ultrasound contrast media (Definity). A destructive pulse was fired first, followed by a chain of non-destructive pulses that allowed for visualization of vascular contrast agent replenishment. Parametric maps of the time required for contrast agent replenishment and the time-integrated intensity were generated for both the tumor and kidney. Following ultrasound examination, contrast-enhanced computed tomography of each tumor was performed in the same imaging plane as that used to acquire the ultrasound images. Fifteen days after the start of treatment, tumors were excised, preserved in 10% formalin, and sectioned in a plane approximating the ultrasound and CT imaging planes. Sections were prepared for light microscopy with H & E, CD31 and factor VIII immunostain to evaluate overall morphology and vessel distribution. Ultrasound measurements of tumor volume, the spatial extent of contrast enhancement, and the time required for contrast replenishment within control tumors were significantly different from those of treated tumors. The time-integrated ultrasound contrast enhancement decreases and the time required for replenishment of the contrast agent within the tumor volume increases over the course of anti-angiogenic therapy. Parametric maps of integrated intensity are shown to correlate with the regions of viable tumor demonstrated on H & E and regions of elevated contrast intensity on CT. Contrast-enhanced ultrasound imaging of implanted tumors provides a tool to assess differences in the microcirculation of treated and control tumors in studies of anti-angiogenic agents. PMID- 12625774 TI - Dynamic breast tumor oximetry: the development of prognostic radiology. AB - A novel pre clinical approach to evaluating tumor oxygen dynamics was recently introduced (Am. J. Clin. Oncol. 24, 462-466 (2001)). FREDOM (Fluorocarbon Relaxometry using Echo planar imaging for Dynamic Oxygen Mapping) allows maps of tumor pO(2) including 50 - 150 individual locations simultaneously to be produced with typical in plane resolution of 1.25 mm in 6.5 mins. The technique has been applied extensively in rat prostate tumors and is now demonstrated in the rat breast 13762NF adenocarcinoma. When anesthetized rats breathed 33% oxygen, mean baseline pO(2) was in the range 17 +/- 2 (se) torr to 74 +/- 4 torr with mean value for nine tumors 46 +/- 8 torr. However, small tumors (< 2.2 cm(3)) were significantly better oxygenated with mean pO(2) = 63 +/- 7 torr than large tumors (> 2.4 cm(3)) with mean pO(2) 24 +/- 5 torr (p < 0.002). Switching the inhaled gas to oxygen or carbogen produced a significant and rapid increase in mean pO(2) for both small and larger tumors (p < 0.05). Given the increasing evidence that tumor oxygenation is related to therapeutic outcome, we believe this approach to measuring tumor oxygen dynamics can be of value in predicting response to therapy, evaluating adjuvant interventions designed to modulate response to therapy, and in providing "Prognostic Radiology". PMID- 12625776 TI - Steady-state and dynamic contrast MR imaging of human prostate cancer xenograft tumors: a comparative study. AB - Understanding tumor vascular physiology is critically important for developing non-invasive, molecularly targeted diagnostic agents and therapies. In this study, using three different human prostate cancer xenografts (MDA PCa 2b, PC3, and LnCap), structural and physiological parameters of neoplastic vasculature and interstitum were explored with a widely available magnetic resonance imaging (MRI) pulse sequence (3D SPGR: spoiled gradient echo). Using dual injection technique employing two T1 contrast agents of different molecular masses (Weissleder, R., Cheng, H. C., Marecos, E., Kwong, K. K., Bogdanov, A., Jr. Eur. J. Cancer 34, 1448-1454 (1998).), steady state (SS) MRI measurements and dynamic contrast agent enhancement (DCE) MRI measurements were simultaneously acquired and analyzed using a two-compartment model for calculating parameters reflecting tumoral architecture and physiology. In particular, interstitial volume and vascular permeability were independently quantified using these two different MRI techniques. Relative vascular water exchange rate, calculated by the flip angle (FA) dependence of measured blood volume using SS technique, and vascular permeability of contrast agent, extrapolated from DCE MRI, were compared. It was found that the SS and DCE techniques were comparable and yielded similar qualitative results for extravascular compartment (interstitial volume). However, the permeability (water exchange rate and contrast agent vascular permeability) values were in disagreement. The results of MR studies are important for interpreting optical imaging results obtained using long-circulating of tumor associated enzymatic activity. PMID- 12625775 TI - Measurement of volumetric and vascular changes with dynamic contrast enhanced MRI for cancer therapy monitoring. AB - Longitudinal dynamic contrast enhanced MRI studies were undertaken to monitor therapy induced volumetric and vascular changes. Three study components are presented in this work: one animal tumor chemotherapy study (R3230 AC adenocarcinoma treated with Taxotere), one patient with invasive lobular breast cancer undergoing neoadjuvant chemotherapy (AC regimen), and one patient with brain metastasis of primary breast cancer undergoing radiation therapy (40 Gray whole brain irradiation). In the animal study two contrast media with different molecular weights, Gadodiamide and Gadomer-17, were used. Only Gadomer-17 revealed significant changes in vascular properties. The responders showed decreased V(b) (vascular volume index) and K(2) (out-flux transport rate), which preceded tumor regression. The control tumors showed increased V(b) and K(2), before tumor growth became much faster. In the patient undergoing neoadjuvant therapy, the tumor was shrinking by 45% after 2 cycles of treatment, then again by 45% after 2 additional cycles. K(2) was decreasing over time with treatment. In the patient with brain metastasis, the 2 follow-up studies were much longer apart to monitor the regression and relapse of lesions. The pre-treatment volumes of lesions in the group without recurrence were significantly smaller compared to those with recurrence. In summary, the tumor volume was more sensitive than the vascular parameters measured by the small extracellular contrast medium for the assessment of therapy response and prediction of recurrence. The vascular properties measured by macromolecular contrast medium may have the potential to serve as early therapeutic efficacy indicators. PMID- 12625779 TI - Oncoproteomics: state-of-the-art. AB - Proteomics is a promising approach in the identification of proteins and biochemical pathways involved in carcinogenesis. Proteomic technologies are now being incorporated in oncology in the post-genomic era. Cancer involves alterations in protein expression and provides a good model not only for detection of biomarkers but also their use in drug discovery. Proteomics has an impact on diagnostics as well as drug discovery. Genomics still remains an important approach but the value of proteomics lies in the fact that most of the diagnostics and drugs target proteins. The importance of application of proteomics in oncology is recognized by the publication of this special issue of TRCT. PMID- 12625777 TI - Congruent MRI and near-infrared spectroscopy for functional and structural imaging of tumors. AB - We present a combined near-infrared diffuse optical spectroscopy (DOS) and Magnetic Resonance Imaging (MRI) system for the study of animal model tumors. A combined broadband steady-state and frequency domain optical spectroscopy apparatus was integrated with the MRI. The physiological properties of tissue rendered by MRI, including vascular volume fraction and water, were compared with chromophore concentrations as determined from the parameters obtained by optical measurements. DOS measurements provided oxy-hemoglobin, deoxy-hemoglobin, and water concentration locally in tumors. A method for co-registration of the information obtained by both modalities was developed. Using Monte Carlo simulations, the optically sampled volume was superimposed on the MR images, illustrating which tissue structure was probed optically. Finally, two optical contrast agents, indocyanine green (ICG) and methylene blue (MB), were employed and their kinetics were measured by DOS system from different locations on the tumor and compared with Gd-DTPA enhancement maps obtained from MRI. PMID- 12625780 TI - Understanding cancer through proteomics. AB - Proteomics is a rapidly expanding discipline that aims to gain a comprehensive understanding of the expressions, modification, interactions, and regulation of proteins in cells. New high-throughput technologies, such as protein chips and isotope-coded affinity tag peptide labeling, coupled with classic technologies such as two-dimensional gel electrophoresis and mass spectrometry, complement genomic technologies, providing cancer researchers with powerful tools for cancer diagnosis and prognosis and for the identification of targets for therapy. PMID- 12625781 TI - Proteomics-based anticancer drug discovery and development. AB - Proteins are important targets for drug discovery and this applied to cancer as well because there is a defect in the protein machinery of the cell in malignancy. Proteomic technologies are now being integrated with genomic approaches for cancer drug discovery and target validation. Among the large number of proteomic technologies available for this purpose, the most important ones are 3-D protein structure determination, protein microarrays, laser capture microdissection and study of protein-protein and protein-drug interactions. Cancer biomarkers and several cell pathways are important drug targets. Several companies are involved in using proteomic technologies for drug discovery. Finally, proteomic approaches will play an important role in the discovery and development of personalized medicines. PMID- 12625782 TI - Contributions of proteome profiling to the molecular analysis of cancer. AB - The proteome is the most functional compartment encoded for in the genome. Technologies for protein separation and quantitation, coupled with mass spectrometry for protein identification, have provided the means for proteome profiling of tumor cell lines and tissues that complement genomic and transcriptomic profiling. The application of established and novel proteomic technologies to the molecular analysis of cancer is reviewed. PMID- 12625783 TI - Application of proteomics in the search for novel proteins associated with the anti-cancer effect of the synthetic cyclin-dependent kinases inhibitor, bohemine. AB - The purpose of this study was to use the proteomics approach, which is based on high resolution two-dimensional electrophoresis coupled with multivariate correspondence analysis and mass spectrometry, to classify objectively the biochemical basis of the anti-cancer activity of the synthetic cyclin-dependent kinase inhibitor, bohemine (BOH). The changes in the cell cycle and corresponding protein composition of the A549 human lung adenocarcinoma cell line after treatment with BOH were evaluated and proteins differentially expressed in the BOH treated A549 cells, compared to the untreated A549 counterparts, were selected. Thirteen of these candidate proteins associated with the drug effects in vitro were identified by mass spectrometry. Many of these proteins fall into one of three functional categories: i) metabolic pathways (glycolysis, nucleic acid synthesis and NADPH production), ii) stress response and protein folding, and iii) cytoskeleton and exocytosis. Changes in protein expression patterns corresponded to a higher resistance of A549 lung carcinoma cells to BOH when compared to the CEM leukaemia cell line. These protein changes reflect a fine balance of the resistant versus the susceptible phenotype in response to the drug. Since BOH is a selective cyclin-dependent kinase inhibitor, changes in the protein expression pattern can be more generally associated with cell cycle regulation as evidenced by inhibition of cell cycling in A549 cells. Our conclusions further underline the importance of cell cycle control in both the cellular signalling and metabolic pathways. PMID- 12625784 TI - Identification of tumor-associated antigens using proteomics. AB - In the post-genomic era, the identification of tumor-associated antigens that elicit a humoral response is allowed at the protein level using proteomics. Indeed, the screening of autoantibodies using 2-D Western blot experiments with sera from cancer patients, followed by the subsequent identification of the target protein by mass spectrometry and database search has permitted the exploitation of the B-cell repertoire of patients with cancer. Applied to several types of cancer, a proteomic-based approach has revealed a high frequency of autoantibodies in sera from patients. Several of the antigenic proteins identified may constitute novel cancer markers and may have clinical utility in diagnosis or in establishing prognosis. Furthermore, the approach has allowed to distinguish isoforms that may help to define epitopes. On the other hand, the analysis of the expression levels of some of the antigenic proteins has revealed differential expression in tumors as compared with healthy tissues that might explain antigenicity. PMID- 12625785 TI - Clinical proteomics for cancer biomarker discovery and therapeutic targeting. AB - As we emerge into the post-genome era, proteomics finds itself as the driving force field as we translate the nucleic acid information archive into understanding how the cell actually works and how disease processes operate. Even so, the traditionally held view of proteomics as simply cataloging and developing lists of the cellular protein repertoire of a cell are now changing, especially in the sub-discipline of clinical proteomics. The most relevant information archive to clinical applications and drug development involves the elucidation of the information flow of the cell; the "software" of protein pathway networks and circuitry. The deranged circuitry of the cell as the drug target itself as well as the effect of the drug on not just the target, but also the entire network, is what we now are striving towards. Clinical proteomics, as a new and most exciting sub-discipline of proteomics, involves the bench-to-bedside clinical application of proteomic tools. Unlike the genome, there are potentially thousands of proteomes: each cell type has its own unique proteome. Moreover, each cell type can alter its proteome depending on the unique tissue microenvironment in which it resides, giving rise to multiple permutations of a single proteome. Since there is no polymerase chain reaction equivalent to proteomics- identifying and discovering the "wiring diagram" of a human diseased cell in a biopsy specimen remains a daunting challenge. New micro-proteomic technologies are being and still need to be developed to drill down into the proteomes of clinically relevant material. Cancer, as a model disease, provides a fertile environment to study the application of proteomics at the bedside. The promise of clinical proteomics and the new technologies that are developed is that we will detect cancer earlier through discovery of biomarkers, we will discover the next generation of targets and imaging biomarkers, and we can then apply this knowledge to patient-tailored therapy. PMID- 12625786 TI - SELDI ProteinChip array in oncoproteomic research. AB - Research into the causes, early detection and treatment of cancers is a primary focus of the health care industry and proteomic-based methodologies are providing an increasingly important role in addressing these issues. The ProteinChip Array technology forms the basis of a clinical proteomics platform designed to expedite the discovery, validation, and characterization of cancer biomarkers at all stages of cancer progression. Being able to detect cancer progression early in turn allows for the possibility of more effective treatment. This short review serves to introduce the technology by highlighting specific examples related to cancer biomarker discoveries. PMID- 12625787 TI - Role of proteomics in diagnosis of cancer. AB - Proteomic technologies have emerged as an important addition to the genomic and antibody-based technologies for the diagnosis of cancer. Important technologies include 2-D gel electrophoresis, mass spectrometry, laser capture microdissection, detection of molecular markers of cancer and protein patterns. For clinical applications, the most likely technologies to be used widely are protein biochips. Application of these technologies to various cancers are described. Proteomic technologies have a potential in developing molecular diagnostics and markers for the early detection of cancer. However, information from various diagnostic technologies should be integrated to obtain the optimal information required for clinical management of a patient. PMID- 12625789 TI - The role of proteomics in the diagnosis and outcome prediction in colorectal cancer. AB - Colorectal cancer is the second most frequent cancer in Western countries. Exogenous factors play a major role in the aetiology of sporadic colorectal cancer representing about 90% of all cases, hereditary cancers accounting for about 10% of patients. Thus, in the large majority of cases, cell dysfunction in CRC results from multiple rather than single, gene interactions. Numerous cellular events and environmental influences modify gene expression or post translational protein modifications. Changes like glycosylation of proteins and lipids which are a common feature in colorectal cancer and influence cancer cell behaviour, cannot be directly detected by genetic studies. Better than genomics studies, functional proteomics studies allow the investigation of environmental factors over time, allowing the monitoring of metabolic responses to various stimuli. However, proteomics studies also have several drawbacks: a) current tools only allow narrow-range analyses, b) identification of proteins of interest remains cumbersome, c) protein studies address multiple compounds of high complexity, d) large amount of proteins are necessary to allow analysis, e) protein research require specific tools, e.g. tagged antibodies, that first have to be developed. Some protein tests are already in application for CRC: a classical prognostic test in colorectal cancer is based on the detection and quantification of a single protein (CEA) in body fluids. Recently, a screening assay based on APC protein truncation test has also been proposed. However, studies linking large protein expression patterns with clinical outcome in colorectal cancer are still in their infancy. To be able to predict occurrence of disease, and treatment outcome, more studies on genotype-phenotype correlations are needed both in sporadic and in hereditary colorectal cancer. PMID- 12625788 TI - Proteomics of breast cancer: outcomes and prospects. AB - Breast cancer is a major public health problem. The identification of new markers to differentiate neoplastic from the normal cells, more thorough understanding of different stages of the pathology, as well as the definition of new therapeutic targets, are all of critical importance. With the completion of human genome sequencing and the introduction of mass spectrometry, combined with protein identification via advanced bioinformatics, proteomics has emerged as a valuable tool for the discovery of new molecular markers. New methods in functional proteomics have also been developed to study the intracellular signaling pathways that underline the development of breast cancer. As illustrated with the examples of fibroblast growth factor-2 and H19, an oncogenic, noncoding mRNA, proteomics have become a powerful approach for deciphering the complex signaling circuitry involved in tumor growth. Breast cancer proteomics have already identified proteins of potential clinical interest (such as the molecular chaperone 14-3-3 sigma) and technological innovations in large scale/high throughput analysis are now ushering in new prospects. PMID- 12625790 TI - Brain metastases from gynecological cancers: factors that affect overall survival. AB - We retrospectively reviewed factors that affected overall survival for patients with gynecological cancers that had metastasized to the brain. Between January 1985 to November 1999, we treated 25 patients with brain metastases from gynecological malignancies (cervix n=6, endometrium n=10, and ovary n=9). Various patient and tumor characteristics were identified and analyzed for their significance. Median age was 46 years old (range, 37-78 years) with the majority of tumors being adenocarcinoma (20/25 patients). The treatment consisted of whole brain radiation therapy (WBRT) in 11 patients, focal therapy (surgery and/or stereotactic radiosurgery [SRS]) in 6 patients, and combination therapy (WBRT and surgery and/or SRS) in 8 patients and resulted in median survivals of 6 months, 7 months and 11 months, respectively. Overall median survival was 7.3 months (range, 1 to 88 months). Cause of death was systemic in 9, neurologic in 8 and progression of primary in 2. Those with single lesions had better median survivals compared to those with multiple lesions (17 months vs. 3 months, p=0.017). Our results suggest that patients with a single lesion had improved outcomes. We encourage enrollment of patients with brain metastases onto prospective clinical trials. PMID- 12625791 TI - Helical tomotherapy: an innovative technology and approach to radiation therapy. AB - Helical tomotherapy represents both a novel radiation treatment device and an innovative means of delivering radiotherapy. The helical tomotherapy unit itself is essentially a hybrid between a linear accelerator and a helical CT scanner for the purpose of delivering intensity-modulated radiation therapy (IMRT). The imaging capacity conferred by the CT component allows targeted regions to be visualized prior to, during, and immediately after each treatment. The megavoltage CT (MVCT) images supplant the port-films used in conventional radiotherapy, providing unprecedented anatomical detail. Image-guidance through MVCT will allow the development and refinement of the concept of "adaptive radiotherapy", the reconstruction of the actual daily delivered dose (as opposed to planned dose) accompanied by prescription and delivery adjustments when appropriate. In addition to this unique feature, helical tomotherapy appears capable of further improvements over 3-dimensional conformal radiation therapy and non-helical IMRT in the specific avoidance of critical normal structures, i.e "conformal avoidance", the counterpart of conformal radiation therapy. Based on radiobiological principles that exploit the physical advantages of helical tomotherapy, several dosimetric and clinical investigations are underway. PMID- 12625792 TI - Effectiveness of antidepressants: comparative remission rates. AB - Historically, clinical researchers have gauged the short-term effectiveness of antidepressants by response rates, which have been defined as a "significant" reduction of symptoms or a global impression of at least moderate benefit. However, increased focus over the past decade has led many researchers to suggest that remission, i.e., a virtual elimination of depressive symptoms and restoration of psychosocial functioning, should be the primary goal of the initial phase of therapy. This article examines the relative efficacy of various antidepressant therapies. There is some evidence that medications affecting multiple neurochemical systems, such as the tricyclics amitriptyline and clomipramine (in studies of hospitalized patients) and the more selective "dual reuptake inhibitor" venlafaxine, may result in higher rates of remission relative to other agents. Given the better tolerability of newer antidepressants relative to tricyclics, both logic and an increasing amount of data support a greater role for multiaction antidepressants. PMID- 12625793 TI - Using treatment algorithms to bring patients to remission. AB - Many patients treated with antidepressants fail to achieve full remission, and the costs, both social and economic, of response without remission as well as residual symptoms are high. Patients who experience incomplete remission to antidepressant treatment are candidates for a sequential treatment approach involving treatment options such as switching, augmentation, or combination of antidepressants. Recently, the number of alternatives for treatment has increased substantially. Algorithms and treatment guidelines that synthesize current data and research provide clinicians with a structure when changes in treatment strategy are necessary. Guidelines and algorithms are not designed to take away the clinician's autonomy but instead are intended to provide support for treatment decisions, and effective ones allow for a wide degree of flexibility. It can be easily argued that the use of algorithms with the associated decision support tools increases the role of the clinician in assessment of the clinical status and subsequent treatment choices. PMID- 12625794 TI - The use of antidepressants in novel combination therapies. AB - Antidepressant monotherapy is used more often than other therapies to achieve symptom remission in depressed patients; however, for patients resistant to antidepressants, other strategies are necessary. Many novel combination therapies have been proposed to treat resistant depression. The efficacy of combination therapies such as lithium augmentation of antidepressants is supported by a large amount of evidence including data from controlled trials. Nonetheless, anecdotal reports suggest that these combinations are underutilized. Data from studies of the use of the combination of atypical antipsychotics and selective serotonin reuptake inhibitors suggest that this is a particularly promising therapeutic avenue. However, more research is needed to corroborate these early results. PMID- 12625795 TI - The role of algorithms in the detection and treatment of depression in primary care. AB - Many depressed patients present to primary care physicians instead of psychiatrists, and several studies have found that primary care physicians underdiagnose and undertreat depression. Primary care physicians may fail to diagnose and treat depression for many reasons: depression as it appears in primary care is in many ways a different disease than that seen in the psychiatric setting, and the process of detecting and treating it is different as well. Primary care clinicians face 2 main tasks when addressing depression in routine practice: to accurately identify those patients who are most likely to benefit from antidepressant treatment and to provide the best treatment options possible in the limited time available per visit. Treatment algorithms can be useful, but they are often difficult to apply in primary care. This article reviews the evidence about the detection and treatment of depression in primary care, examines current research on the differences between the primary and specialty care environments, and discusses aspects of treatment guidelines and algorithms that are important to primary care physicians. PMID- 12625796 TI - Treating generalized anxiety disorder. AB - Generalized anxiety disorder (GAD) is characterized by chronically persistent worry and therefore requires effective long-term treatment. This article reviews the benefits and risks associated with various pharmacologic and psychological therapies to assess their ability to achieve the elimination of GAD symptomatology and restoration of normal function. Psychotherapeutic approaches such as applied relaxation, cognitive therapy, and cognitive-behavioral therapy have all been shown to be effective when used as monotherapies and may be beneficial when used adjunctively. Current effective pharmacotherapies for patients with GAD include anxiolytic benzodiazepines, buspirone, and antidepressants including venlafaxine and paroxetine. Benzodiazepines have long been used to treat anxiety and are particularly appropriate in short-term treatment situations; however, their adverse side-effect profile and their inability to treat depression commonly comorbid with GAD renders them less than ideal in many situations. Buspirone has demonstrated anxiolytic benefits but, like benzodiazepines, shows negligible antidepressant action. Antidepressants like paroxetine and venlafaxine are not only effective antidepressants but also effective anxiolytics, thus implying their special ability to treat GAD and concurrent depression, even over the long-term. PMID- 12625797 TI - Use of algorithms to treat anxiety in primary care. AB - The presentations of anxiety in primary care are more numerous and broader in spectrum than the presentations of depression, and the primary care physician is often faced with the challenge of teasing out a diagnosis from the full spectrum of anxiety disorders. A treatment algorithm that begins with recognition and diagnosis and carries the primary care physician and patient through long-term treatment and, finally, withdrawal of treatment can be a useful and appropriate tool. Use of an algorithm targeted specifically for primary care physicians treating patients with anxiety disorders would insure that patients in the primary care setting receive the best care during treatment of anxiety disorders, while primary care physicians become better able to serve a broader community. PMID- 12625798 TI - New approaches to managing difficult-to-treat depressions. AB - In a sense, all depression is difficult to treat. Most depressions are episodic conditions that, not infrequently, are slow to fully remit. Most also are complicated by comorbid psychiatric and general medical disorders. However, a minority of such difficult-to-treat depressions are treatment resistant. The most common cause of initial treatment failure is not resistance but undertreatment that is, an insufficient duration of treatment, a subtherapeutic dosage of antidepressant, and/or poor adherence to the prescribed regimen. Complicating factors such as undiagnosed hypothyroidism or substance abuse can result in apparent treatment resistance unless addressed. Challenging subtypes of illness, including psychotic and bipolar subtypes of depression, are not necessarily inherently refractory but must be met with modified treatment approaches. PMID- 12625799 TI - Managing treatment-resistant major depression. AB - A sizable proportion of patients suffering from nonpsychotic unipolar depression experience only partial or no clinical response to antidepressant treatment. Switching, augmenting, and combining various pharmaceutical agents can be effective strategies for patients with treatment-resistant depression. The empirical evidence supporting these approaches is inconsistent, however, and there is a paucity of controlled studies to support their efficacy. Additionally, it has been difficult to demonstrate the advantages of these strategies over increasing the dose or duration of the initial drug treatment. This article will review available evidence and clinical considerations regarding switching, augmenting, and combining various agents in the treatment of patients suffering from nonpsychotic unipolar depression who have failed adequate courses of antidepressant treatment. More research is needed that controls for continued time on the initial agent, that compares different strategies, and that determines which patients are the best candidates for which treatment. PMID- 12625800 TI - New approaches in managing bipolar depression. AB - Historically, the pharmacologic treatment of bipolar depression has not been well studied. New data are beginning to emerge regarding the efficacy of new medications and the use of combinations of mood stabilizers and antidepressants in acute and long-term treatment of bipolar depression. We reviewed data from recent randomized, controlled trials of mood stabilizers and antidepressants in the treatment of bipolar depression and naturalistic studies examining the risk of switching and depressive relapse with ongoing antidepressant treatment. PMID- 12625801 TI - New approaches to managing psychotic depression. AB - Major depression with psychotic features, while fairly common, is frequently misdiagnosed. Symptoms seen in these patients are those of an overall severe depressive disorder with psychomotor impairment (retardation or agitation), guilt, suicidal preoccupation, and neuropsychological impairment. A number of biological characteristics and behavioral symptoms are specific to patients suffering from psychotic depression and differ significantly from those of nonpsychotic depression. Psychotic depression is seen in patients of all ages, and it has a high short-term morbidity and risk of suicide. Data support the use of antipsychotics in combination with antidepressants for major depression with psychotic features, but other treatments may have as great or greater efficacy for the disorder. This article focuses on recognizing the features of psychotic depression, the success of current treatment options, and new treatments under investigation. PMID- 12625802 TI - Difficult-to-treat depressions: a primary care perspective. AB - Depression is common in primary care and more difficult to treat than many clinicians are aware. The goal of treatment is symptomatic remission, and by current estimates 50% or more of patients treated with antidepressant monotherapy may suffer from residual neurovegetative, cognitive, and somatic symptoms. Bipolar disorder, in particular, is more prevalent in primary care than previously recognized, is easily misdiagnosed, and may be a significant source of treatment failure. This article reviews treatment resistance, its causes, and management approaches. Many strategies are straight-forward and within the skill set of primary care clinicians. The use of antidepressants with multiple mechanisms of action may reduce first-order resistance. Antidepressant augmentation strategies (e.g., with lithium or atypical antipsychotics) are often very effective and readily instituted by informed and motivated practitioners. PMID- 12625803 TI - Neurohumoral control of gastrointestinal motility. AB - Neurohumoral substances and their receptors play a major part in the complex regulation of gastrointestinal motility and have therefore been the predominant targets for drug development. The numerous receptors involved in motility are located mainly on smooth muscle cells and neuronal structures in the extrinsic and intrinsic parts of the enteric nervous system. Within this system, receptor agonists and antagonists interacts directly to modify excitatory or inhibitory signals. In view of this complexity it is not surprising that our knowledge about the mechanisms of actions of the various neurohormones and drugs affecting gut motility has been rather fragmented and incomplete. However, recently substantial progress has been achieved, and drug therapy for gut dysmotility is emerging, based primarily on neurohumoral receptors. This paper presents a selective review of the neurohumoral regulatory mechanisms of gastrointestinal motility. In this context, the physiology and pharmacology of the smooth muscle cells, gastrointestinal motility and dysmotility, the enteric nervous system, gastrointestinal reflexes, and serotonin is presented. Further investigation and understanding of the transmitters and receptors involved in especially the reflex activation of peristalsis is crucial for the development of novel therapies for motility disorders. PMID- 12625804 TI - Enteral nutrition and hepatosplanchnic region in critically ill patients - friends or foes? AB - Enteral nutrition (EN) is a preferred way of feeding in critically ill patients unless obvious contraindications such as ileus or active gastrointestinal bleeding are present. Early enteral nutrition as compared to delayed EN or total parenteral nutrition decreases morbidity in postsurgical and trauma patients. The hepatosplanchnic region plays a pivotal role in the pathophysiology of sepsis and multiple organ dysfunction syndrome. The beneficial effects of EN on splanchnic perfusion and energy metabolism have been documented both in healthy volunteers and animal models of sepsis, hemorrhagic shock and burns. By contrast, EN may increase splanchnic metabolic demands, which in turn may lead to oxygen and/or energy demand/supply mismatch, especially when hyperemic response to EN is not preserved. Therefore, the timing of initiation and the dose of EN in patients with circulatory failure requiring vasoactive drugs are a matter of controversy. Interestingly, the results of recent clinical studies suggest that early enteral nutrition may not be harmful even in patients with circulatory compromise. Nevertheless, possible onset of serious complications, the non-occlusive bowel necrosis in particular, have to be kept in mind. Unfortunately, there is only a limited number of clinically applicable monitoring tools for the effects of enteral nutrition in critically ill patients. PMID- 12625805 TI - Statistics in three biomedical journals. AB - In this paper we analyze the use of statistics and associated problems, in three Czech biological journals in the year 2000. We investigated 23 articles Folia Biologica, 60 articles in Folia Microbiologica, and 88 articles in Physiological Research. The highest frequency of publications with statistical content have used descriptive statistics and t-test. The most usual mistake concerns the absence of reference about the used statistical software and insufficient description of the data. We have compared our results with the results of similar studies in some other medical journals. The use of important statistical methods is comparable with those used in most medical journals, the proportion of articles, in which the applied method is described insufficiently is moderately low. PMID- 12625806 TI - Inhibin B, follicle stimulating hormone, luteinizing hormone and testosterone during childhood and puberty in males: changes in serum concentrations in relation to age and stage of puberty. AB - Inhibin B is a gonadal dimeric polypeptide hormone that regulates synthesis and secretion of follicle stimulating hormone (FSH) in a negative feedback loop. The aim of the present study was to determine changes in serum inhibin B, gonadotropins and testosterone concentrations during childhood and puberty in males. We studied the relationship between circulating inhibin B, gonadotropins and testosterone in serum of healthy boys during the first two years of life and then in pubertal development. Using a recently developed two-side enzyme-linked immunosorbent assay (ELISA), inhibin B levels were measured in the serum of 78 healthy boys divided into eleven age groups from birth to the end of pubertal development. In addition, serum levels of gonadotropins and testosterone were measured. Serum inhibin B, gonadotropins and testosterone increased during the first months of postnatal life. A peak in serum inhibin B and gonadotropins concentrations was observed around 3-4 months of age. There was a significant positive correlation between serum inhibin B and gonadotropins and testosterone levels during the first 2 years of life. After this early increase, serum inhibin B, gonadotropins and testosterone levels decreased significantly and remained low until puberty followed by an increase beginning with the onset of puberty. Serum levels of inhibin B reached a peak at stage G3 of puberty. Around midpuberty, inhibin B lost its positive correlation with luteinizing hormone (LH) and testosterone from early puberty, and developed a strong negative correlation with FSH, which persisted into adulthood. We conclude that inhibin B plays a key role in the regulation of the hypothalamic-pituitary-gonadal hormonal axis during male childhood and pubertal development. Inhibin B is a direct marker of the presence and function of Sertoli cells and appears to reflect testicular function in boys. PMID- 12625807 TI - Leptin levels in obese children: effects of gender, weight reduction and androgens. AB - Obesity in children is accompanied by increased circulating leptin concentrations. Girls have higher leptin concentrations than boys. The aim of our study was to compare serum leptin levels before and after a five-week weight reduction program and to study the relationship of leptin levels, serum total cholesterol, and androgens (testosterone, dehydroepiandrosterone, dehydroepiandrosterone sulphate) in 33 obese boys (age: 12.7+/-1.97 years, BMI: 30.46+/-4.54) and 66 obese girls (age: 12.7+/-2.51 years, BMI: 29.31+/-4.62). We found that serum leptin concentrations in obese children were significantly decreased after a weight reduction program (before 20.79+/-9.61 ng/ml, after 13.50+/-8.65 ng/ml in girls; before 12.25+/-10.09 ng/ml and after 5.18+/-3.56 ng/ml in boys, p<0.0001 in both genders). Leptin levels correlated positively with the body mass index before and after weight reduction. There was a positive association in obese boys and a negative one in obese girls between leptin levels and the WHR (waist to hip circumference ratio). Serum leptin also shows a strong relationship to fat distribution (p=0.02 in boys, p<0.0001 in girls). No significant correlation was found between leptin concentrations and total cholesterol or androgens. We confirmed that leptin is a sensitive parameter of body composition and weight reduction in obese children. PMID- 12625808 TI - Serum ghrelin levels in obese patients: the relationship to serum leptin levels and soluble leptin receptors levels. AB - Ghrelin is a new endogenous ligand for the growth hormone secretagogue receptor. It activates the release of growth hormone from the pituitary and it also participates in the regulation of energy homeostasis. The aim of the study was to characterize changes in serum ghrelin levels in obese subjects and their relationship to the serum levels of leptin and soluble leptin receptor. Eight obese patients (6 women and 2 men) with body mass index (BMI) 40.3+/-13.4 kg.m( 2) and eight healthy controls (5 women and 3 men) with BMI 22.7+/-1.3 kg.m(-2) were examined. The ghrelin serum levels (165.0+/-58.1 vs. 343.37+/-81.96; p<0.001) and soluble leptin receptor serum levels (7.25+/-3.44 vs. 21.80+/-4.99; p<0.0001) were significantly lower in obese patients. The leptin serum levels (23.45+/-12.90 vs. 6.41+/-2.96; p<0.005) were significantly higher compared to the lean subject group. In both measured groups the levels of serum leptin significantly positively correlated with BMI. We proved a significantly lower serum ghrelin levels in the group of obese patients in comparison with the control group. PMID- 12625809 TI - Different activation of ACTH and corticosterone release in response to various stressors in rats. AB - The aim of this study was to investigate the reaction of the hypothalamo pituitary-adrenocortical (HPA) system to various stressors (fasting, crowding, cold and heat) by measuring blood ACTH and corticosterone (CORT) concentration as well as the cholesterol (CHOL) content in the adrenals. To examine the effects of stress termination, the rats were returned and kept under control conditions for the same period as that of stress duration (supposed recovery period). According to our results HPA system was activated by all the stressors applied. Heat seems to be the strongest stressor since the exposure of animals to a high ambient temperature resulted in the greatest rise of plasma ACTH concentration as well as CORT synthesis and secretion. These values remained elevated after the stress termination i.e. after the rats had been returned to room temperature. Fasting seems to be the weakest stressor given because it causes the smallest increase in blood ACTH and CORT concentrations. Moreover, in refed rats the HPA function was fully recovered. In conclusion, the various stressors applied seem to induce a different response of the HPA system as judged by quantitative changes in ACTH and CORT release. PMID- 12625810 TI - Hepatotoxic effect of D-galactosamine and protective role of lipid emulsion. AB - D-galactosamine is a hepatotoxic agent, which induces diffuse injury of liver tissue followed by the regeneration process. Our data showed a high increase of serum aminotransferases after D-galactosamine administration, which indicates a high extent of liver injury. When lipid emulsion was applied immediately after D galactosamine, the increase of serum aminotransferases was greatly reduced. In addition, the decrease of the cytochrome c oxidase activity induced by D galactosamine was not observed after lipid emulsion administration and the increase of total liver oxidative capacity in the regeneration period due to activated mitochondrial biogenesis was accelerated. All these findings indicate a protective effect of lipid emulsion administration against D-galactosamine toxicity. PMID- 12625811 TI - IgA and IgG antigliadin, IgA anti-tissue transglutaminase and antiendomysial antibodies in patients with autoimmune thyroid diseases and their relationship to thyroidal replacement therapy. AB - Celiac disease is a chronic illness of the small bowel caused by gliadin intolerance in genetically predisposed subjects. The aim of this study was to investigate serum levels of IgA and IgG antigliadin antibodies, IgA antiendomysial antibodies, and IgA anti-tissue transglutaminase antibodies in 169 patients with autoimmune thyroid diseases, i.e. chronic thyroiditis and Graves' disease. Antiendomysial antibodies were positive in 2 out of 169 persons (1.18%), IgA antigliadin antibodies in 15.98%, IgG antigliadin antibodies in 51.48%, and IgA anti-tissue transglutaminase in 14.79%. The prevalence of positivity was higher compared to the 1312 control blood donors described in our previous study (Vancikova et al. 2002) (p<0.05). Patients with chronic thyroiditis treated with a high replacement dosage of levothyroxin (125-200 microg daily) had higher serum levels of IgA antigliadin antibodies in comparison with patients treated with a lower dosage (50-100 microg daily) (medians: 13.00 vs. 19.69, p=0.033). We found a negative correlation of IgA anti-tissue transglutaminase antibodies and total calcium serum levels (r = -0.480, p=0.0236, n=22). We can conclude that in persons with autoimmune thyropathy there is a high prevalence of positive antigliadin, anti-tissue transglutaminase and antiendomysial antibodies. Latent celiac disease may lead to impaired resorption of therapeutically administered levothyroxine, calcium, or other substances. PMID- 12625812 TI - Changes in acute phase proteins after anti-tumor necrosis factor antibody (infliximab) treatment in patients with Crohn's disease. AB - Acute phase proteins and markers of proteosynthetic activity reflect the clinical activity in Crohn's disease (CD). The impact of anti-tumor necrosis factor antibody (anti-TNF) therapy on serum levels of acute phase proteins and proteosynthetic markers was studied. Fourteen patients with active CD were treated with 5 mg per kg of anti-TNF in intravenous infusion. Clinical activity (assessed by Crohn's disease activity index - CDAI), alpha-1-acid glycoprotein, haptoglobin, cholinesterase and prealbumin were assessed before and in months 1 and 5 after treatment. A sustained decrease in CDAI was observed. This was accompanied by a significant decrease in alpha-1-acid glycoprotein and haptoglobin in month 1 (p=0.005 and p=0.01, respectively) while in month 5 the levels of both acute phase proteins rose significantly (p=0.003 for alpha-1-acid glycoprotein and p=0.02 for haptoglobin). Cholinesterase and prealbumin significantly increased in month 1 after the treatment (p=0.02 and p=0.0006, respectively), the increase was sustained in cholinesterase while prealbumin levels diminished in month 5. We conclude that the clinical improvement after anti-TNF therapy for CD is accompanied by changes of acute phase proteins and proteosynthetic markers. The assessment of these laboratory markers may be useful in the management of CD patients treated with anti-TNF. PMID- 12625813 TI - Different patterns of serum interleukin 10 response to treatment with anti-tumor necrosis factor alpha antibody (infliximab) in Crohn's disease. AB - Administration of anti-tumor necrosis factor antibody (anti-TNF, infliximab) down regulates T helper 1 (Th 1) cytokines production in intestinal mucosa of patients with Crohn's disease (CD). Interleukin 10 (IL-10) is thought to be involved in CD pathogenesis through regulation of the Th 1 response. The aim of this study was to determine the IL-10 response in CD patients treated with anti-TNF. Fourteen patients with active CD received 5 mg/kg of infliximab; clinical activity assessed by Crohn's Disease Activity Index (CDAI), alpha1-acid glycoprotein and serum IL-10 were determined before and after treatment, in month 0, 1 and 5. In the group with a good clinical response, IL-10 levels diminished significantly in month 1 (p<0.05) and remained decreased in month 5. The group with a lower response showed a significant increase in IL-10 levels in month 1 (p<0.05). alpha1-acid glycoprotein levels obtained before treatment were significantly elevated in the group with a good clinical response (p<0.05) and a significant decrease in month 1 was observed in this group (p<0.05). These observations suggest that a pattern of IL-10 response might be related to the clinical response to anti-TNF treatment in CD. PMID- 12625814 TI - Changes in the superoxide production and other macrophage functions could be related to the mortality of mice with endotoxin-induced oxidative stress. AB - Free radicals and proinflammatory cytokines from phagocytes have been implicated in the pathogenesis of endotoxic shock, a disease with high mortality caused by Gram-negative bacterial endotoxin. In the present study, male BALB/c and Swiss mice received intraperitoneally lipopolysaccharide (LPS) at 100 mg/kg and 150 mg/kg, respectively, that led to a lethal endotoxic shock (100 % of mortality before 30 h). Swiss mice injected with 100 mg/kg, that did not show lethal endotoxic shock, were also studied. Peritoneal macrophages were obtained from animals at 2, 4, 12 or 24 h after injection of LPS or saline (control) solutions. Superoxide anion and tumor necrosis factor (TNFalpha) production were determined in these cells as well as other functions such as adherence capacity, chemotaxis and phagocytosis. The increase in superoxide anion production after endotoxin injection was higher in cells from mice with lethal shock than in those with non lethal shock. However, the enhancement of TNFalpha production was similar in all cases, although in Swiss mice the highest levels of TNFalpha were observed at 1.5 h after endotoxin injection, while in BALB/c mice they occurred at 2 h after LPS injection. This oxidative stress was also revealed by the other functions analyzed, since adherence to substrate and phagocytosis were stimulated and chemotaxis was decreased after endotoxin injection as compared to controls, the differences being even more significant in animals with lethal shock. These data suggest that these changes, mainly the increased production of free radicals even more than the TNFalpha release, could be involved in mouse mortality caused by LPS. PMID- 12625815 TI - The effect of hyperbaric oxygen treatment on oxidative stress in experimental acute necrotizing pancreatitis. AB - Various protocols may be used for acute pancreatitis treatment. Recently, the benefit of hyperbaric oxygen (HBO) has been demonstrated. To clarify the mechanism of HBO on the process of the acute pancreatitis, we determined the levels of antioxidant enzymes in an acute pancreatitis model. Forty-five Sprague Dawley rats were randomly divided into three groups: Group I: sham group (n=15), Group II: pancreatitis group (n=15), Group III: pancreatitis group undergoing HBO therapy (n=15). HBO was applied postoperatively for 5 days, two sessions per day at 2.5 fold absolute atmospheric pressure (ATA) for 90 min. Superoxide dismutase (Cu/Zn-SOD), malondialdehyde (MDA), and glutathione peroxidase (GSH Px) activity were measured in pancreatic tissue and erythrocyte lysate. MDA and GSH Px were also determined in plasma. In addition, amylase levels were measured in the serum. While serum amylase levels and MDA values in erythrocyte, plasma and pancreatic tissue were decreased, the levels of GSH Px and SOD were found to be significantly increased in the Group III as compared to those of the Group II. The findings of our study suggest that HBO has beneficial effects on the course of acute pancreatitis and this effect may occur through the antioxidant systems. PMID- 12625816 TI - The onset of apoptosis of neurons induced by ischemia-reperfusion injury is delayed by transient period of hypertension in rats. AB - We investigated the potential neuroprotective effect of transient hypertension on neuronal cell death induced by ischemia-reperfusion. Recovery of neurons, terminally differentiated cells, is almost entirely dependent upon active transcription and repair of DNA damage. We focused on the histochemical detection of distribution of NOR (argyrophylic nucleolar proteins) reflecting nucleolar integrity, immunohistochemical detection of PARP-1 (poly(ADP-ribose) polymerase 1), MADD (mitogen-activated death domain), a protein accumulated in nucleoli upon stimulation by ischemia, the active form of caspase-3, a universal proteolytic enzyme of apoptosis. The terminal deoxynucleotidyl-transferase (TdT)-mediated dUTP-biotin nick-end-labeling method (TUNEL) proved the presence of in situ DNA fragmentation. We used the model of transient focal cerebral ischemia in rats with occlusion of middle cerebral artery. In experimental group of rats, the transient hypertension was induced by constriction of the abdominal aorta. The period of ischemia lasted 15, 30, 60 and 120 min followed by 48 h of reperfusion. We examined the frontal lobe of the ipsilateral hemisphere for apoptosis of neurons and compared it with the intact brain tissue. In normotensive rats with transient focal cerebral ischemia, we found disintegrated nucleoli of cortical as well as subcortical neurons at all investigated periods of ischemia, whereas the neurons of intact animals showed compact nucleoli with a few satellites. Nuclear positivity for MADD and PARP-1 was apparent in the neocortex after 15 min and peaked after 30 min of ischemia. On the other hand, the subcortical neurons showed nuclear positivity after 60 and 120 min. The immunohistochemical reaction for active caspase 3 was apparent after 30 min onwards predominantly in the cortex. The TUNEL staining was distinct after 60 and 120 min. In hypertensive rats, we found nucleolar disintegration, positivity for MADD, PARP-1 and caspase 3 after 30 min cortically and subcortically, followed by TUNEL positive staining of cortical neurons after 60 and 120 min. In summary, we detected delayed activation of neuronal apoptosis in transiently hypertensive rats with focal cerebral ischemia compared to normotensive animals. The apoptotic phenotype was confirmed by a panel of complementary methods showing rapid proteolysis-nucleolar segregation, MADD, PARP-1 and caspase-3 positivity as well as ultimate DNA fragmentation proved by the TUNEL assay. PMID- 12625817 TI - Actions of lyotropic anions on the mechanical properties of fast and slow twitch rat muscles at different temperatures. AB - The effects of lyotropic (swelling) anions (Cl(-), Br(-), NO(3)(-) and I(-)) on contractile properties of fast-twitch extensor digitorum longus (EDL) and slow twitch soleus (SOL) muscles were investigated in vitro at 20 degrees C and 35 degrees C. Isolated muscles bathed in anionic Tyrode solution were stimulated directly and isometric single twitches and fused tetanic contractions were recorded. In a Cl(-)Tyrode solution a decrease of the bathing temperature led to a cold potentiation of the twitch tension (P(t)) in EDL muscles, however, to a cold depression in SOL muscles, in both muscles combined with a prolongation of contraction (CT) and half relaxation (HRT) times. The extent and order of the potentiating effect of lyotropic anions on the P(t), CT and HRT in EDL and SOL were quite similar and increased in the order: Cl(-)< Br(-)< NO(3)(-)< I(-). Since the lyotropic anions did not influence tetanic tensions, the twitch-tetanus ratio (TTR) was increased in NO(3)(-) and I(-)solutions. All effects of the anions were rapidly and completely reversed in both muscles when the test solution was replaced by the normal one. The temperature decrease caused no significant alteration in the potentiation capacity of the anions or in the kinetics of their action and reversibility. PMID- 12625819 TI - Heart rate and increased intravascular volume. AB - The objective was to establish whether an intravascular volume increase leads to a heart rate (HR) increase without increased sympathetic tonus. HR changes at rest and at deep breathing (6/min - simulated increase of atrial filling pressure) were measured in patients after heart transplantation. Evaluation of dependency of HR changes on breathing depth was done through a new time series methodology. The data was evaluated through graphs displaying a significant increase in the graph area at deep breathing, when compared with breathing at rest (p<0.01). We presume that an increase in HR corresponds to increased intravascular volume and malfunctioning kidneys. PMID- 12625818 TI - Oxidative stress in the brain tissue of laboratory mice with acute post insulin hypoglycemia. AB - Malondialdehyde (MDA), Cu,Zn-superoxide dismutase (Cu,Zn-SOD) and selenium dependent glutathione peroxidase (GSPHx) are currently considered to be basic markers of oxidative stress. MDA is one of the end-products of the peroxidation of membrane lipids, whereas enzymes Cu,Zn-SOD and GSHPx belong to the natural antioxidants. The role of oxygen free radicals in the pathogenesis of many diseases is well documented. The aim of this study was to ascertain the influence of insulin-induced acute hypoglycemia on oxidative stress in the brain tissue. Hypoglycemia was induced in ICR mice by intraperitoneal administration of insulin at a dose 24 IU/kg. There was a correlation between the severity of hypoglycemia and the levels of MDA, Cu,Zn-SOD and GSHPx. The results showed that in severe hypoglycemia (serum glucose concentration below 1.0 mmol/l) the lipoperoxidation in brain tissue expressed as the level of MDA was higher in comparison with normoglycemic controls (glycemia around 3.7 mmol/l) as well as in comparison with the levels of MDA during moderate hypoglycemia (glycemia ranging between 1-2 mmol/l). This indicates the enhancement of lipoperoxidation in the brain tissue during severe hypoglycemia. However, both enzymes - Cu,Zn-SOD or GSHPx - did not show a similar tendency. PMID- 12625820 TI - Novel "three-in-one" peptide device for genetic drug delivery. AB - We here describe a new strategy for the delivery of oligonucleotides to cells that is based on the use of a short peptide containing three functional units: a membrane-penetrating segment, a DNA-binding domain and a cell-localization sequence. The designed vector binds strongly to oligonucleotides and has membrane perturbing abilities in vitro. This type of multi-functional device may be a powerful tool to achieve efficient delivery of genetic drugs in vivo. PMID- 12625821 TI - The temperature dependence of gramicidin conformational States in octanol. AB - In lipid bilayers and organic solvents, the hydrophobic polypeptide gramicidin adopts a number of different conformations, some of which are capable of conducting monovalent cations across phospholipid membranes. The equilibria between conformations have been shown to be influenced by factors such as lipid chain length, solvent, concentration and salt. In this study, the temperature dependence of the equilibrium mixture of double helical ion-free gramicidin in octanol was examined using circular dichroism spectroscopy. PMID- 12625823 TI - pH-induced conformational change in an alpha-helical coiled-coil is controlled by His residues in the hydrophobic core. AB - An alpha-helical coiled-coil structure is one of the basic structural units in proteins. Hydrophilic residues at the hydrophobic positions in the coiled-coil structure play important roles in structures and functions of natural proteins. We reported here a peptide that formed a triple stranded alpha-helical coiled coil showing the pH-dependent structural change. The peptide was designed to have two His residues at the hydrophobic positions of the center of the coiled-coil structure. The peptide folded into a triple stranded coiled-coil at neutral pH, while it unfolded at acidic pH. This construct is useful to create a protein that the structure or function is controlled by pH. PMID- 12625822 TI - Steady-state cleavage kinetics for dengue virus type 2 ns2b-ns3(pro) serine protease with synthetic peptides. AB - The N-terminal part of the NS3 protein from dengue virus contains a trypsin-like serine protease responsible for processing the nonstructural region of the viral polyprotein. Enzymatic activity of the NS2B-NS3(pro) precursor incorporating a full-length NS2B cofactor of dengue virus type 2 was examined by using synthetic dodecamer peptide substrates encompassing native cleavage sequences of the NS2A/NS2B, NS2B/NS3, NS3/NS4A and NS4B/NS5 polyprotein junctions. Cleavage of the dansylated substrates was monitored by a HPLC-based assay and kinetic parameters for K(1M), k(cat) and k(cat)/K(m) were obtained. The data presented here show that NS2B-NS3(pro) expressed in recombinant E. coli can be renatured to an active protease which reacts in the absence of microsomal membranes with all 4 substrate peptides, albeit the molecule does not exhibit autoproteolytic processing at the NS2B/NS3 site. A marked difference in cleavage efficiency was found for the NS2B/NS3 substrate and the remaining 3 peptides based on the NS2A/NS2B, NS3/NS4A and NS4A/NS5 cleavage sites. PMID- 12625824 TI - Inhibition of human napsin A. AB - The newly-discovered human aspartic proteinase, napsin A was not susceptible to protein inhibitors from potato, squash or yeast but was weakly inhibited by the 17 kDa polypeptide from Ascaris lumbricoides and potently by isovaleryl and lactoyl-pepstatins. A series of synthetic inhibitors was also investigated which contained in the P(1)-P(1)' positions the dipeptide analogue statine or its phenylalanine or cyclohexylalanine homologues and in which the residues occupying P(4)-P(3)' were varied systematically. On this basis, the active site of napsin A can be readily distinguished from other human aspartic proteinases. PMID- 12625825 TI - Direct observation of structure transition of donor-acceptor labeled peptides temperature dependence of fluorescence quenching kinetics. AB - From the temperature dependence of fluorescence quenching kinetics, a novel temperature-sensitive structure transition was discovered in a donor-acceptor labeled peptides. Perfect agreement of related abrupt changes in a(1),tau(1) and tau(2) gives strong support for the existence of this structure transition, which suggests it is possible to develop peptides that can be rapidly and reversibly switched between two structure states in response to temperature shift. PMID- 12625826 TI - Kinetic and site-directed mutagenesis studies of prevotella intermedia acid phosphatase. AB - Site-directed mutagenesis was used to examine the roles of the conserved histidine, arginine and cysteine residues in acid phosphatase from Prevotella intermedia (PiACP). The replacement of histidine and arginine residues resulted in the elimination of the PiACP activity while the cysteine mutants retained activity. These results suggest that the histidine and arginine residues are essential for catalysis. PMID- 12625827 TI - A structure-function analysis of glial cell-line-derived neurotrophic factor receptor alpha1. AB - The GFRalpha1 cDNA was amplified by RT-PCR from fetal rat hippocampus. The soluble recombinant GFRalpha1 and its mutants were obtained from an Escherichia coli expression system. The biological activity of soluble GFRalpha1 and its mutants were evaluated in PC12 cells. The results suggest that the central domain of GFRalpha1 is a crucial determinant for ligand binding. This established a solid basis for further study to find the key amino acid mediating the binding of GDNF and GFRalpha1. PMID- 12625828 TI - The denaturation of alpha, beta and psi bovine trypsin at pH 3.0: evidence of intermediates. AB - The conformational stability and the folding process of alpha, beta and Psi bovine trypsin at pH 3.0 followed by circular dichroism (CD) and size exclusion in HPLC have been analyzed as a function of urea concentration. The thermodynamic stability for a and b are deltaG = 15.91 +/- 0.28 kcal/mol, deltaG = 15.54 +/- 2.39 kcal/mol. respectively, and y trypsin is deltaG = 16.10 +/- 2.51 kcal/mol. The transition curves for alpha, beta and Psi forms suggest a molten globule state. PMID- 12625829 TI - N-terminal domain unfolds first in the sequential unfolding of papain. AB - Temperature and Guanidine hydrochloride induced unfolding transitions of papain at pH 2.0 are biphasic implying independent and sequential unfolding of its two domains. To determine the order of unfolding, the active site located in the interface of the domains was labeled with an environment specific fluorescent probe (1,8-IAEDANS). Unfolding of this complex relative to the free protein followed by intrinsic and extrinsic fluorescence measurements suggests that the N domain unfolds initially in the sequential unfolding of domains. PMID- 12625830 TI - Low content of protein S29 in ribosomes of human lung cancer cell line a549: detected by two-dimensional electrophoresis. AB - This study revealed that the content of protein S29 in ribosomes of cancer cell line A549 was distinctly low (equivalent to about 30% of that of 2BS). The conclusion was acquired based on the ratios of spot volume of ribosomal protein S29 to that of several other ribosomal proteins (S29/L37a, S29/L38, S29/S27 and S29/S28) in the same gel plate. The possible biological roles of ribosomal protein S29 in malignant transformation and translation regulation are briefly discussed. PMID- 12625831 TI - Spectroscopic analysis of the stability of bothrops myotoxic phospholipases A2 to guanidine and urea denaturation. AB - Spectrophotometric profiles representing the unfolding induced by guanidine on Bothrops moojeni myotoxins-I (MjTX-I) and II (MjTX-II), Bothrops jararacussu bothropstoxin-I (BthTX-I) and Bothrops pirajai piratoxin-I (PrTX-I) were obtained and compared with those obtained with bovine ribonuclease A (RNAse) and trypsin. The molar (epsilon(1M)) and percent (epsilon(1%)) extinction coefficients were determined for the four myotoxins as well as for RNAse and trypsin as reference parameters. These coefficients were then used throughout this work. The changes in free energy (deltaGD(H)(2)(O)) corresponding to zero guanidine concentration and the guanidine concentrations (D(1/2)) able to convert 50% of the molecules from the native to the unfolded state were determined. The values of deltaGD (H)(2)(O) ranged from 4.42 (BthTX-I) to 8.02 (MjTX-I) kcal/mole, compared with 6.47 and 6.88 kcal/mole for trypsin and RNAse, respectively. The values for deltaGD(H)(2)(O) and D1/2 showed that BthTX-I is the least stable among the four myotoxins assayed, with a D1/2 close to that of RNAse, while MjTX-II is conformationally the most stable. Monitoring of the unfolding of RNAse and PrTX-I by a 0 to 6 M urea gradient PAGE revealed transitions from the native (N) to the unfolded (U) state with deltaG(N-U)of 0.22 and 0.41 kcal/mole, respectively. Sigmoidal curves showed well-defined two-stage transitions for both proteins. PMID- 12625832 TI - The effect of a mixed meal on endothelium-dependent vasodilation is dependent on fat content in healthy humans. AB - Impaired endothelium-dependent vasodilation (EDV) is an early marker of atherosclerosis. The aim of the present study was to investigate how meals with different fat contents influence endothelial vasodilatory function. A total of 26 young, healthy men and women aged 20-30 years ingested an ordinary Western meal [34 energy% (E%) fat, n =10], or isocaloric meals with low-fat (20 E%, n =8), or minimal-fat (3 E%, n =8) content. EDV was assessed as forearm blood flow (FBF) during local administration of 4 microg/min methacholine chloride (Mch-FBF) and endothelium-independent vasodilation as FBF during administration of 10 microg/min sodium nitroprusside (SNP-FBF) at baseline and 1 and 2 h after each meal. FBF was determined by venous occlusion plethysmography. An endothelial function index (EFI) was calculated as the Mch-FBF/SNP-FBF ratio. Both Mch-FBF and the EFI were decreased at 1 h after the 34 E% fat meal ( P <0.01 and P <0.05 respectively), but approached fasting levels after 2 h. Mch-FBF and EFI did not change significantly in the group consuming the 20 E% fat meal, but increased in the 3 E% fat group ( P <0.01 and P <0.05 compared with baseline for Mch-FBF and EFI respectively). SNP-FBF was not significantly affected by any of the meals. In conclusion, low-fat meals did not attenuate EDV, in contrast with an ordinary Western meal, which transiently impaired EDV. Our findings indicate that a dietary fat content of 20 E% or less might be beneficial to endothelial vasodilatory function. PMID- 12625833 TI - Oestrogen replacement therapy lowers plasma levels of asymmetrical dimethylarginine in healthy postmenopausal women. AB - Oestrogen replacement therapy (ERT) has been shown to lead to favourable changes in the cardiovascular risk profile of postmenopausal women. Part of this effect is ascribed to increased production or bioavailability of nitric oxide (NO). We have tested the hypothesis that ERT lowers plasma levels of asymmetrical dimethylarginine (ADMA), an endogenous inhibitor of NO synthase (NOS). In a randomized double-blind study design, 40 hysterectomized postmenopausal women received conjugated equine oestrogen (CEE; 0.625 mg/day; n =14), the selective oestrogen receptor modulator raloxifene (150 mg/day; n =13) or placebo ( n =13). At baseline and after 6, 12 and 24 months of treatment, plasma was analysed for levels of arginine, ADMA, and symmetrical dimethylarginine (SDMA), a stereoisomer of ADMA that does not inhibit NOS. An overall treatment effect on ADMA levels was observed in the CEE group ( P =0.004 compared with placebo), but not in the raloxifene group ( P =0.50). The decrease of ADMA levels by CEE treatment was consistent over the 2-year study period, without significant differences between the effects at 6, 12 and 24 months. The average post-baseline change in ADMA in the CEE group compared with placebo was -7.8% (95% confidence interval -12.8% to 2.9%; P =0.003). Arginine or SDMA levels did not change during treatment in any of the groups. Thus ERT with oral conjugated oestrogen, but not with raloxifene, significantly reduced plasma concentrations of the cardiovascular risk factor ADMA in healthy postmenopausal women. PMID- 12625834 TI - Identification of S1 proteins B2, C1 and D1 as AUF1 isoforms and their major role as heterogeneous nuclear ribonucleoprotein proteins. AB - AUF1 (A+U-rich RNA binding factor) participates in the rapid decay of mRNAs in the cytoplasm. It is sometimes called heterogeneous nuclear ribonucleoprotein (hnRNP) D0; however, evidence for its characterization as an hnRNP protein has been scarce. S1 proteins A-D are those selectively extracted at pH 4.9 from isolated nuclei pretreated with either RNase A or DNase I. In the present study we identified S1 ('first supernatant') proteins B2, C1 and D1 with p45, p40 and p37 AUF1s respectively, by microsequencing and product analysis of transfected cDNAs. We found, further, that more than 96% of the S1 proteins occurred in the nucleus, and localized largely in RNase-sensitive structures. B2 was confined in the nucleus and C1 directly bound to heterogeneous nuclear RNAs (hnRNAs). These B2 and C1 proteins formed hnRNP structures responsible for the 33 S, and, to lesser extent, the 40 S particles, which were liberated upon mild nucleolytic cleavage. On the other hand, D1 and the remainder of C1 were associated with nuclease-hypersensitive sites of hnRNAs, and comprised the major cytoplasmic AUF1s that may be involved in mRNA decay. Two-dimensional immunoblotting resolved each S1 isoform into up to six spots or more, and suggested that the previous uncertain relationship of hnRNP D0 and hnRNP D is resolved in terms of charge differences and differential splicing arising from one gene. The present results thus indicate that S1 proteins B2, C1 and D1 are identical with AUF1 proteins, but largely occur as hnRNP proteins in the nucleus. That hnRNP D0 is indeed an hnRNP protein was verified. PMID- 12625836 TI - New insights into the possible role of bacteriophages in host defense and disease. AB - BACKGROUND: While the ability of bacteriophages to kill bacteria is well known and has been used in some centers to combat antibiotics - resistant infections, our knowledge about phage interactions with mammalian cells is very limited and phages have been believed to have no intrinsic tropism for those cells. PRESENTATION OF THE HYPOTHESIS: At least some phages (e.g., T4 coliphage) express Lys-Arg-Gly (KGD) sequence which binds beta3 integrins (primarily alphaIIbbeta3). Therefore, phages could bind beta3+ cells (platelets, monocytes, some lymphocytes and some neoplastic cells) and downregulate activities of those cells by inhibiting integrin functions. TESTING THE HYPOTHESIS: Binding of KGD+ phages to beta3 integrin+ cells may be detected using standard techniques involving phage - mediated bacterial lysis and plaque formation. Furthermore, the binding may be visualized by electron microscopy and fluorescence using labelled phages. Binding specificity can be confirmed with the aid of specific blocking peptides and monoclonal antibodies. In vivo effects of phage - cell interactions may be assessed by examining the possible biological effects of beta3 blockade (e.g., anti-metastatic activity). IMPLICATION OF THE HYPOTHESIS: If, indeed, phages can modify functions of beta3+ cells (platelets, monocytes, lymphocytes, cancer cells) they could be important biological response modifiers regulating migration and activities of those cells. Such novel understanding of their role could open novel perspectives in their potential use in treatment of cardiovascular and autoimmune disease, graft rejection and cancer. PMID- 12625835 TI - Thermotoga neapolitana adenylate kinase is highly active at 30 degrees C. AB - The adenylate kinase (AK) gene from Thermotoga neapolitana, a hyperthermophilic bacterium, was cloned and overexpressed in Escherichia coli, and the recombinant enzyme was biochemically characterized. The T. neapolitana AK (TNAK) sequence indicates that this enzyme belongs to the long bacterial AKs. TNAK contains the four cysteine residues that bind Zn(2+) in all Gram-positive AKs and in a few other Zn(2+)-containing bacterial AKs. Atomic emission spectroscopy and titration data indicate a content of 1 mol of Zn(2+)/mol of recombinant TNAK. The EDTA treated enzyme has a melting temperature (T (m)=93.5 degrees C) 6.2 degrees C below that of the holoenzyme (99.7 degrees C), identifying Zn(2+) as a stabilizing feature in TNAK. TNAK is a monomeric enzyme with a molecular mass of approx. 25 kDa. TNAK displays V (max) and K (m) values at 30 degrees C identical with those of the E. coli AK at 30 degrees C, and displays very high activity at 80 degrees C, with a specific activity above 8000 units/mg. The unusually high activity of TNAK at 30 degrees C makes it an interesting model to test the role of enzyme flexibility in activity. PMID- 12625837 TI - Mechanomyography versus electromyography, in monitoring the muscular fatigue. AB - BACKGROUND: The use of the mechanomyogram (MMG) which detects muscular vibrations generated by fused individual fiber twitches has been refined. The study addresses a comparison of the MMG and surface electromyogram (SEMG) in monitoring muscle fatigue. METHODS: The SEMG and MMG were recorded simultaneously from the same territory of motor units in two muscles (Biceps, Brachioradialis) of the human (n = 18), during sustained contraction at 25 % MVC (maximal voluntary contraction). RESULTS: The RMS (root mean square) of the SEMG and MMG increased with advancing fatigue; MF (median frequency) of the PSD (power density spectra) progressively decreased from the onset of the contraction. These findings (both muscles, all subjects), demonstrate both through the SEMG and MMG a central component of the fatigue. The MF regression slopes of MMG were closer to each other between men and women (Biceps 1.55%; Brachialis 13.2%) than were the SEMG MF slopes (Biceps 25.32%; Brachialis 17.72%), which shows a smaller inter-sex variability for the MMG vs. SEMG. CONCLUSION: The study presents another quantitative comparison (MF, RMS) of MMG and SEMG, showing that MMG signal can be used for indication of the degree of muscle activation and for monitoring the muscle fatigue when the application of SEMG is not feasible (chronical implants, adverse environments contaminated by electrical noise). PMID- 12625838 TI - Dynamics of a disabled population in Morocco. AB - BACKGROUND: The disabled population constitutes a class of people needing special care and necessitating important economic and social effort. METHODS: In this paper, using specific parameter settings, partial differential equations are used to model the temporal change of the proportion of the disabled population in Morocco. RESULTS: Combining different forms and values of the parameters, a numerical method is proposed and three scenarios are considered. These forms and values are determined by data fitting and simulation. CONCLUSIONS: The experiments show clearly the dynamical evolution of the disabled population with time and age according to each scenario. PMID- 12625839 TI - Assessment of diffuse Lewy body disease by 2-[18F]fluoro-2-deoxy-D-glucose positron emission tomography (FDG PET). AB - BACKGROUND: Lewy body disease is, after Alzheimer's disease, the second most common cause of senile degenerative dementia with progressive cognitive deterioration, fluctuation of cognitive and motoric functions and psychotic symptoms. It is characterized histologically by the occurrence of Lewy bodies in allocortical, neocortical and subcortical structures. The aim of this study was to measure the cortical glucose metabolism using FDG PET (2-[18F]fluoro-2-deoxy-D glucose position emission tomography) compared to normal subjects. PATIENTS AND METHODS: Five patients (5 m, mean age 75 y) with clinically suspected diffuse Lewy body disease (DLB) were studied with FDG PET. PET studies of the head were performed with a Siemens ECAT-ART PET-scanner with attenuation correction using 137-Cs point sources. RESULTS: We found the same distribution pattern of diffuse glucose hypometabolism in the entire cortical region with relative sparing of the primary sensory-motor cortex in all the patients. The few cases reported in the literature so far describe findings similar to ours. CONCLUSION: The pattern of diffuse glucose hypometabolism in the entire cortex including the occipital region seems to be a typical feature of DLB that is distinctive from dementia of Alzheimer's disease. PMID- 12625840 TI - Transfection of IL-10 expression vectors into endothelial cultures attenuates alpha4beta7-dependent lymphocyte adhesion mediated by MAdCAM-1. AB - BACKGROUND: Enhanced expression of MAdCAM-1 (mucosal addressin cell adhesion molecule-1) is associated with the onset and progression of inflammatory bowel disease. The clinical significance of elevated MAdCAM-1 expression is supported by studies showing that immunoneutralization of MAdCAM-1, or its ligands reduce inflammation and mucosal damage in models of colitis. Interleukin-10 (IL-10) is an endogenous anti-inflammatory and immunomodulatory cytokine that has been shown to prevent inflammation and injury in several animal studies, however clinical IL 10 treatment remains insufficient because of difficulties in the route of IL-10 administration and its biological half-life. Here, we examined the ability of introducing an IL-10 expression vector into endothelial cultures to reduce responses to a proinflammatory cytokine, TNF-alpha METHODS: A human IL-10 expression vector was transfected into high endothelial venular ('HEV') cells (SVEC4-10); we then examined TNF-alpha induced lymphocyte adhesion to lymphatic endothelial cells and TNF-alpha induced expression of MAdCAM-1 and compared these responses to control monolayers. RESULTS: Transfection of the IL-10 vector into endothelial cultures significantly reduced TNF-alpha induced, MAdCAM-1 dependent lymphocyte adhesion (compared to non-transfected cells). IL-10 transfected endothelial cells expressed less than half (46 +/- 6.6%) of the MAdCAM-1 induced by TNF-alpha (set as 100%) in non-transfected (control) cells. CONCLUSION: Our results suggest that gene therapy of the gut microvasculature with IL-10 vectors may be useful in the clinical treatment of IBD. PMID- 12625843 TI - Erectile dysfunction: an overview. AB - Sexual function is a central part of a man's identity, helping to define who he is, and how he feels about himself. It is also a key determinant of relationship health. Erectile dysfunction (ED) threatens all of this, and is thus an important topic in male health. Erectile dysfunction is common around the world, especially in older men. Risk factors for ED overlap significantly with those for cardiovascular disease and endothelial dysfunction, especially diabetes. PMID- 12625844 TI - A historical review of erectile dysfunction. AB - Over the last three decades, there has been a significant increase in our understanding of the physiologic mechanisms responsible for erectile dysfunction. Erectile dysfunction has become a topic of considerable media and societal interest and acceptance. Paralleling the increase in knowledge has been an explosion in therapeutic options. This article will evaluate the therapeutic options, from a historical perspective of what has been available, and outline the progress that has been made. PMID- 12625841 TI - New knowledge from old: in silico discovery of novel protein domains in Streptomyces coelicolor. AB - BACKGROUND: Streptomyces coelicolor has long been considered a remarkable bacterium with a complex life-cycle, ubiquitous environmental distribution, linear chromosomes and plasmids, and a huge range of pharmaceutically useful secondary metabolites. Completion of the genome sequence demonstrated that this diversity carried through to the genetic level, with over 7000 genes identified. We sought to expand our understanding of this organism at the molecular level through identification and annotation of novel protein domains. Protein domains are the evolutionary conserved units from which proteins are formed. RESULTS: Two automated methods were employed to rapidly generate an optimised set of targets, which were subsequently analysed manually. A final set of 37 domains or structural repeats, represented 204 times in the genome, was developed. Using these families enabled us to correlate items of information from many different resources. Several immediately enhance our understanding both of S. coelicolor and also general bacterial molecular mechanisms, including cell wall biosynthesis regulation and streptomycete telomere maintenance. DISCUSSION: Delineation of protein domain families enables detailed analysis of protein function, as well as identification of likely regions or residues of particular interest. Hence this kind of prior approach can increase the rate of discovery in the laboratory. Furthermore we demonstrate that using this type of in silico method it is possible to fairly rapidly generate new biological information from previously uncorrelated data. PMID- 12625842 TI - Glycosaminoglycans in human retinoblastoma cells: heparan sulfate, a modulator of the pigment epithelium-derived factor-receptor interactions. AB - BACKGROUND: Pigment epithelium-derived factor (PEDF) has binding affinity for cell-surface receptors in retinoblastoma cells and for glycosaminoglycans. We investigated the effects of glycosaminoglycans on PEDF-receptor interactions. RESULTS: 125I-PEDF formed complexes with protease-resistant components of medium conditioned by human retinoblastoma Y-79 cells. Using specific glycosaminoglycan degrading enzymes in spectrophotometric assays and PEDF-affinity chromatography, we detected heparin and heparan sulfate-like glycosaminoglycans in the Y-79 conditioned media, which had binding affinity for PEDF. The Y-79 conditioned media significantly enhanced the binding of 125I-PEDF to Y-79 cell-surface receptors. However, enzymatic and chemical depletion of sulfated glycosaminoglycans from the Y-79 cell cultures by heparitinase and chlorate treatments decreased the degree of 125I-PEDF binding to cell-surface receptors. CONCLUSIONS: These data indicate that retinoblastoma cells secrete heparin/heparan sulfate with binding affinity for PEDF, which may be important in efficient cell-surface receptor binding. PMID- 12625845 TI - Pharmacology of phosphodiesterase 5 inhibitors. AB - The phosphodiesterase enzymes, of at least 11 types, are ubiquitous throughout the body, and perform a variety of functions. Phosphodiesterase type 5 (PDE5) is the predominant enzyme in the corpus cavernosum, and plays a crucial role in penile erection. Inhibitors of PDE5 are the most effective oral agents in the treatment of erectile dysfunction. Sildenafil, tadalafil, and vardenafil are all potent inhibitors of PDE5 and show the same mechanism of action, although they have some pharmacological differences that may translate into varying clinical effects. PMID- 12625846 TI - Tadalafil: a new agent for erectile dysfunction. AB - Oral phosphodiesterase 5 (PDE5) inhibitors for the treatment of erectile dysfunction are preferred by most men, and are recommended in guidelines as first line therapy, because of convenience, high efficacy, and low rates of side effects. Tadalafil (Cialis) is a new agent that has been studied in different patient populations. It has a different molecular structure than other PDE5 inhibitors, and a different pharmacologic profile that provides a longer period of effectiveness than other agents. This article will review clinical trials on tadalafil, to provide a comprehensive overview of its efficacy and safety. PMID- 12625847 TI - Cardiovascular safety of PDE5 inhibitors. AB - Erectile dysfunction (ED) and cardiovascular disease share many of the same risk factors and have some common elements of pathophysiology. Clinically, they often coexist. Another link between the two conditions is that sildenafil, the first oral therapeutic agent effective in treating ED, has been shown to potentiate the hypotensive effects of nitrates, a potentially serious side effect. Nitrates are commonly used in the treatment of coronary artery disease. As such, sildenafil (and, likely, other new phosphodiesterase type 5 [PDE5] inhibitors) is contraindicated in men who use nitrate medications. This article will examine the risk of an acute coronary event during sexual activity, and review an algorithm for evaluating the cardiac risk of a patient with ED. The interaction between PDE5 inhibitors and cardiac medications will be discussed, along with guidelines for using sildenafil in men with cardiac disease. PMID- 12625850 TI - Reforming Canada's medical care system. PMID- 12625851 TI - A meaningful legacy: urologists as Nobel Prize laureates. AB - OBJECTIVE: To review the careers of two urologists among Nobel Prize-winners in medicine, W. Forssmann and C. H. Huggins, and the significance of their contributions. MATERIAL AND METHODS: Investigation was performed based on analysis of collected findings from the biographies of laureates, their scientific publications and the Nobel archive database. RESULTS: Review revealed that of the 175 scientists and physicians who received the Nobel Prize, just over one half (94) held an MD degree while the remainder were PhD's or other degrees. Of the 94 MD-degreed physicians nine (9.4%) were surgeons. Two of these laureates were urologists- Drs. Werner Forssmann and Charles B. Huggins, who were awarded the Nobel Prize in 1956 and 1966, respectively. Although Werner Forssmann worked as a urologist for most of his career, early in his surgical training he invented procedures for cardiac catheterization and performed the first procedures on himself in 1929. Charles Huggins identified the role of androgens in prostate cancer progression in 1940, and thus established the principles of hormonal suppressive therapy for advanced disease. CONCLUSIONS: The distinguished accomplishments of these two great urologists exemplify the highest level of excellence in science for the entire surgical and urological community. Furthermore, today's breakthroughs in molecular medicine represent an extremely appealing challenge for the new generation of scientists and clinicians. PMID- 12625852 TI - Radical prostatectomy: size of the prostate gland and its relationship with acute perioperative complications. AB - OBJECTIVE: To determine if there is correlation between the size of radical prostatectomy specimens and perioperative complications including intraoperative blood loss. METHODS: One hundred twenty consecutive retropubic radical prostatectomy cases were retrospectively reviewed. Perioperative complications, intraoperative blood loss, pathologic stage, and size of the prostatectomy specimen were recorded. Logistic regression was used to determine whether variables such as age, PSA, and prostate weight are significant predictors of perioperative complications and intraoperative blood loss. RESULTS: The final analysis included a total of 117 cases. Significant complications were seen in 10 patients (8.5%). The median weight of the prostatectomy specimen in the group with major complications was 44.5 g (range 24-219) which was significantly higher than the median weight of 39.9 g (range 13-124) for the group without any complications (p = 0.034). The size of the prostate gland predicted the likelihood of a perioperative complication better than chance. A prostate size greater than 37 g was 10 times more likely to encounter major complications. Our analysis also indicated a statistically significant positive correlation between the weight of the prostatectomy specimen and intraoperative blood loss (p = 0.046). CONCLUSIONS: Prostate size correlates with a higher risk of major perioperative complications and higher intraoperative blood loss. PMID- 12625853 TI - Radical prostatectomy stabilizes peak urinary flow rates. AB - OBJECTIVE: A community-based study of 2,119 men in Olmsted County reported that median peak urinary flow rates (PFR)s and voided volumes decrease steadily after age 40. We wanted to study how removal of the prostate via radical prostatectomy (RRP) would effect age-related reduction of PFRs and voided volumes. METHODS: One hundred men 1-9 years following RRP were evaluated for PFR, voided volume, post void residual, and AUA symptom score. RESULTS: Following removal of the prostate the mean PFR was 26.6 +/- 11 cc/sec and the mean voided volume was 345 +/- 174 cc. Analysis of PFR and voided volume versus age were stable over all ages. PFR and voided volume versus follow-up demonstrated a flat or slightly positive slope. PFRs post RRP were higher compared to the community-based data that reported mean PFRs that dropped steadily for men in their 50's, 60's and 70's (25.8 to 18.6 cc/sec, 26.3 to 16.1 cc/sec, 27.3 to 13.8 cc/sec, respectively p<.01). CONCLUSION: Following RRP, PFRs are, as expected, higher than historic controls and stable with advancing age and follow-up. This is in contrast to diminishing PFRs seen in normal men as they age. Our study suggests that age related reduction in PFR and voided volumes is largely eliminated after total removal of the prostate gland. PMID- 12625854 TI - A comparative analysis of rapid urine tests for the diagnosis of upper urinary tract malignancy. AB - OBJECTIVES: To compare the effectiveness of two rapid urine tests fibrinogen/fibrin degradation products FDP (Aura Tek FDP, PerImmune Inc., Rockville, Maryland, USA) and bladder tumor antigen BTA (Bard BTA, Bard Canada Inc., Mississauga, Ontario, Canada) to urinary cytology in establishing the diagnosis of transitional cell carcinoma (TCC) in patients with suspected upper tract malignancy. MATERIALS AND METHODS: In a prospective study, urine samples were collected from 29 patients with abnormalities of the upper tracts highly suspicious for malignancy. Sensitivity and specificity of the BTA and FDP tests were determined and compared to those of cytology. All persons interpreting the individual tests were blinded to the other test results. RESULTS: Of the 29 patients evaluated, 14 were found to have upper tract TCC. The overall sensitivity of FDP, BTA and urinary cytology was found to be 100%, 50% and 29%, respectively. The accuracy of the FDP test was 83% as compared to 62% for BTA and 59% for cytology. CONCLUSION: Urinary cytology lacks sensitivity in the diagnosis of upper tract TCC. This preliminary study suggests that the point-of-care test, FDP, has better test characteristics than urinary cytology. Further evaluation of these tests is warranted for their potential to improve and facilitate the diagnosis of upper tract tumors. PMID- 12625855 TI - Comparison of recovery from postoperative pain utilizing two sling techniques. AB - PURPOSE: Bone anchors are used for suture fixation in a wide variety of reconstructive surgeries. They have been in use for pelvic floor reconstruction since 1992. Bone anchors provide a stable point of suture fixation in order to avoid tying over the mobile rectus fascia. The purpose of this study was to compare two sling techniques that utilize bone anchors with respect to recovery from postoperative pain, complete continence, operative time, and length of hospital stay. MATERIALS AND METHODS: A total of 64 women (mean age = 57) were treated for stress urinary incontinence secondary to intrinsic sphincter deficiency or hypermobility between March 1998 to August 2000. Group I (SPWS) consisted of 30 patients who underwent insitu vaginal wall sling with suprapubic placement of bone anchors in the pubic tubercle utilizing the Vesica system. Group II (TVCS) consisted of 34 patients who underwent cadaveric fascia sling with transvaginal placement of bone anchors behind the symphysis pubis utilizing the Precision-TAC system. Phone interviews were conducted by a third party who was blinded to the details of the surgical technique, to assess pain at various postoperative times as well as current level of continence. The pain assessment was done using the Verbal Pain Assessment Scale (VAS). Complete continence was defined as dryness with no pad use. RESULTS: Significant differences were discovered in both days to pain free state and operative time. No other differences were detected in continence or length of hospital stay. Based on the VAS, a pain free state was achieved for the TVCS group in 1.33 days and for the SPWS group in 9.7 days with p=0.00043. Mean operative time for the SPWS group was 96.9 minutes for the sling alone and 106.7 minutes when combined with cystocele repair. Mean operative time for the TVCS group was 75.36 minutes for the sling alone and 98.11 minutes when combined with cystocele repair. No patient in either group developed osteomyelitis, osteitis pubis, removal of the bone anchors for any reason, nor sling erosion. Seventy percent and 83.3% patients were completely dry (mean follow-up 12.5 months, range 3-30 months) in the SPWS and TVCS group, respectively. CONCLUSION: A pain free state is achieved faster in patients undergoing transvaginal placement of bone anchors compared to bone anchors placed suprapubically. Bone anchors used in sling procedures are safe and achieve acceptable short term continence rates. PMID- 12625856 TI - The unsuspected nonpalpable testicular mass detected by ultrasound: a management problem. AB - Ultrasound is recognized as a valuable method of detecting testicular masses. Rarely, ultrasound will detect a testicular mass that was not clinically suspected. We present the case of a 43-year old man who presented with an unsuspected testicular mass detected by ultrasound. He underwent inguinal orchiectomy. The pathology showed a Leydig Cell tumor with cytological atypia. A review of the literature suggests that most incidentally discovered testicular masses are tumors, but there is disagreement as to whether they are usually benign or malignant. In view of the fact that many of these lesions are benign, if tumor markers are not elevated, inguinal exploration and excision of the lesion with frozen section examination is an acceptable management approach. PMID- 12625857 TI - The unsuspected nonpalpable testicular mass detected by ultrasound: a management problem. PMID- 12625858 TI - Group A streptococcal hydrocele infection and sepsis in a renal transplant recipient. AB - A case of hydrocele infection secondary to a cutaneous b-hemolytic group A streptococcal infection is described in a renal transplant recipient. Sepsis and renal failure occurred in the setting of this severe, life-threatening infection. This case represents the first description of a group A streptococcal hydrocele infection in an adult. This type of infection can progress rapidly to sepsis and its attendant complications, especially in an immunocompromised patient. Early diagnosis and treatment is crucial in order to optimize the outcome. PMID- 12625859 TI - Metastatic adenocarcinoma in testis presenting as a testicular mass--a case report and review of literature. AB - Testicular metastasis presenting as a testicular mass is an extremely rare condition. There are only nine previously reported cases where testicular mass was the first clinical manifestation of underlying malignancy. Here we report a case of metastatic mucin secreting adenocarcinoma in testis presenting as a testicular mass with unknown primary. We have given a brief review of literature about the spread of tumor to testes. PMID- 12625860 TI - Vaso-cutaneous fistula after vasectomy. AB - Vaso-cutaneous fistulae occur rarely after vasectomy. We report three cases encountered in our hospital over the last 18 years and recommend technical considerations to avoid this complication. PMID- 12625862 TI - Predictive and correlative techniques for the design, optimisation and manufacture of solid dosage forms. AB - There is much interest in predicting the properties of pharmaceutical dosage forms from the properties of the raw materials they contain. Achieving this with reasonable accuracy would aid the faster development and manufacture of dosage forms. A variety of approaches to prediction or correlation of properties are reviewed. These approaches have variable accuracy, with no single technique yet able to provide an accurate prediction of the overall properties of the dosage form. However, there have been some successes in predicting trends within a formulation series based on the physicochemical and mechanical properties of raw materials, predicting process scale-up through mechanical characterisation of materials and predicting product characteristics by process monitoring. Advances in information technology have increased predictive capability and accuracy by facilitating the analysis of complex multivariate data, mapping formulation characteristics and capturing past knowledge and experience. PMID- 12625863 TI - Modified microplex vector enhances transfection of cells in culture while maintaining tumour-selective gene delivery in-vivo. AB - A non-commercial liposome (dimethyl dioctadecyl ammonium bromide:dioleoyl phosphatidylethanolamine) was compared with a commercial variety (Lipofectamine) for transfection of cultured rat adenocarcinoma cells and in an in-vivo kidney tumour model. Transfection of the cells in culture and in tumours in-vivo was variable with both types of liposomes. A high-dose microplex (lipoplex microsphere) vector enhanced liposome-mediated transfection of cells in culture. When these high-dose microplexes were tested in-vivo, they were better than both microspherical and liposomal delivery modes in terms of transgene expression levels and the tumour-to-normal tissue ratio of gene delivery. Microplexes have been demonstrated to be capable of not only selective delivery of plasmids to solid tumours, but also of increasing transfection in cell culture, a finding that may be used in ex-vivo transfection studies. It is hypothesized that microspheres anchored the combination vector closer to the cultured cells, allowing attached liposomes to gain easier access into cells. In-vivo, microspheres permitted the microplexes to selectively deliver their genetic payload within the tumour tissue, from where the action of cationic liposomes on cellular membranes facilitated increased access of plasmids into the cytosol of target cells. PMID- 12625864 TI - Daily transdermal administration of selegiline to guinea-pigs preferentially inhibits monoamine oxidase activity in brain when compared with intestinal and hepatic tissues. AB - Selegiline has been formulated in an acrylic polymer adhesive mixture to be employed as a constant release topical patch for daily transdermal administration. Application of this selegiline transdermal system (STS) to guinea pigs resulted in an average delivery of 1.185 mg selegiline/cm(2) patch/24 h. STS dose-response curves were generated by altering patch size (cm(2)). A transdermal dose range was identified which inhibited guinea-pig brain monoamine oxidase-B (MAO-B) by greater than 95% yet provided for a dose-dependent inhibition of monoamine oxidase-A (MAO-A) activity. The ID50 for inhibition of MAO-A activity in response to a 21-day daily regimen with transdermal selegiline was approximately 7.5-fold lower for cortical and striatal brain regions compared with that obtained for duodenum; hepatic MAO-A was unaffected following the same dosing regimen. By contrast, orally administered selegiline inhibited brain and duodenal MAO-A to the same extent, and generated a shallower dose-inhibition curve for brain MAO-A inhibition. In addition, transdermal delivery was approximately 6-8-times more potent than oral selegiline for the inhibition of brain MAO-A activity. It is concluded that daily transdermal selegiline administration may provide therapeutic advantages over oral treatment, based on its preferential, dose-dependent inhibition of brain vs peripheral MAO-A activity. PMID- 12625865 TI - Physical stability and enthalpy relaxation of drug-hydroxypropyl methylcellulose phthalate solvent change co-precipitates. AB - The poorly water-soluble drug GWX was co-precipitated with hydroxypropyl methylcellulose phthalate (HPMCP) using a solvent change method. The two co precipitate formulations made, with drug-HPMCP ratios of 2:8 and 5:5, were analysed using modulated temperature differential scanning calorimetry. They were found to consist of completely amorphous solid solution and a mixture of amorphous solid solution, crystalline drug and amorphous drug, respectively. Stability with respect to crystallization of the two co-precipitates and pure amorphous drug made by quench cooling was compared by storing preparations at 25 degrees C and 40 degrees C, under vacuum over P(2)O(5), and at 75% relative humidity (r.h.). Humidity (75% r.h. compared with dry) had a larger influence on crystallization of the amorphous drug than temperature (25 degrees C compared with 40 degrees C). The solid solution phase in co-precipitates had a relatively higher stability than amorphous drug alone, with respect to crystallization, in presence of the plasticizer water, and crystalline drug. These findings were partly explained by evidence of decreased molecular mobility in the amorphous solid solution with respect to amorphous drug alone, using enthalpy relaxation measurements. At an ageing temperature of 65 degrees C, the calculated half-life for enthalpy relaxation of the 2:8 drug-HPMCP ratio coprecipitate was about 6 orders of magnitude greater than that of amorphous drug alone, indicating a large difference in relative molecular mobility. PMID- 12625866 TI - Characterisation and evaluation of novel surfactant bacterial anti-adherent coatings for endotracheal tubes designed for the prevention of ventilator associated pneumonia. AB - It is accepted that ventilator-associated pneumonia is a frequent cause of morbidity and mortality in intensive care patients. This study describes the physicochemical properties of novel surfactant coatings of the endotracheal tube and the resistance to microbial adherence of surfactant coated endotracheal tube polyvinylchloride (PVC). Organic solutions of surfactants containing a range of ratios of cholesterol and lecithin (0:100, 25:75, 50:50, 75:25, dissolved in dichloromethane) were prepared and coated onto endotracheal tube PVC using a multiple dip-coating process. Using modulated temperature differential scanning calorimetry it was confirmed that the binary surfactant systems existed as physical mixtures. The surface properties of both surfactant-coated and uncoated PVC, following treatment with either pooled human saliva or phosphate-buffered saline (PBS), were characterised using dynamic contact angle analysis. Following treatment with saliva, the contact angles of PVC decreased; however, those of the coated biomaterials were unaffected, indicating different rates and extents of macromolecular adsorption from saliva onto the coated and uncoated PVC. The advancing and receding contact angles of the surfactant-coated PVC were unaffected by sonication, thereby providing evidence of the durability of the coatings. The cell surface hydrophobicity and zeta potentials of isolates of Staphylococcus aureus and Pseudomonas aeruginosa, following treatment with either saliva or PBS, and their adherence to uncoated and surfactant-coated PVC (that had been pre-treated with saliva) were examined. Adherence of S. aureus and Ps. aeruginosa to surfactant-coated PVC at each successive time period (0.5, 1, 2, 4, 8 h) was significantly lower than to uncoated PVC, the extent of the reduction frequently exceeding 90%. Interestingly, the microbial anti-adherent properties of the coatings were dependent on the lecithin content. Based on the impressive microbial anti-adherence properties and durability of the surfactant coating on PVC following dip coatings, it is proposed that these systems may usefully reduce the incidence of ventilator-associated pneumonia when employed as luminal coatings of the endotracheal tube. PMID- 12625867 TI - Influence of route of administration on the absorption and disposition of alpha-, gamma- and delta-tocotrienols in rats. AB - A study was conducted to evaluate the bioavailability of alpha-, gamma- and delta tocotrienols administered via oral, intravenous, intramuscular and intraperitoneal routes in rats. Three separate experiments, each conducted according to a two-way crossover design, were carried out to compare intravenous and oral, intramuscular and oral, and intraperitoneal and oral administration. Oral absorption of all three tocotrienols was found to be incomplete. Of the three tocotrienols, alpha-tocotrienol had the highest oral bioavailability, at about 27.7+/-9.2%, compared with gamma- and delta-tocotrienols, which had values of 9.1+/-2.4% and 8.5+/-3.5%, respectively. Such biodiscrimination was also observed in their total clearance rates (estimated from the intravenous data). alpha-Tocotrienol showed the lowest clearance rate at about 0.16 L kg(-1) h(-1), whereas that of delta- and gamma-tocotrienols was quite similar, with values of 0.24 and 0.23 L kg(-1) h(-1), respectively. Interestingly, all three tocotrienols were found to be negligibly absorbed when administered intraperitoneally and intramuscularly. Thus, these two routes of administration should be avoided when evaluating the biological activities of the tocotrienols in whole animal experiments. PMID- 12625868 TI - Gene expression of CYP3A4, ABC-transporters (MDR1 and MRP1-MRP5) and hPXR in three different human colon carcinoma cell lines. AB - Colon carcinoma cell lines are used widely as screening models for intestinal absorption of drugs. However, the expression of important transport systems and of metabolic enzymes is not completely characterized yet. The expression and inducibility of multidrug resistance gene 1 (MDR1) and cytochrome P450 isoform 3A4 (CYP3A4) was investigated in Caco-2 parental, Caco-2 TC-7 (TC-7) and LS180 cell lines. In the same three cell lines, we investigated the expression of isoforms of the multidrug resistance associated protein family (MRP1-MRP5) and the human pregnane X receptor (hPXR), which may be important for MDR1 and CYP3A4 induction. Cells were treated with rifampicin or 1alpha,25 dihydroxycholecalciferol (1,25(OH)(2)D(3)) for 72 h and the total RNA was extracted. Afterwards reverse transcription real-time polymerase chain reaction (TaqMan) assay was performed to determine the mRNA expression level. We have shown that in LS180 cells, MDR1 and CYP3A4 were inducible with both inducers. In Caco-2 parental and TC-7 cells, CYP3A4 was only inducible with 1,25(OH)(2)D(3). Furthermore, differences were shown in gene expression of several transport proteins (MDR1 and MRP1-MRP5) and CYP3A4 in different human colon carcinoma derived cell lines. hPXR mRNA was expressed in all three cell lines but the amount of mRNA detected was significantly higher in LS180 cells than in Caco-2 and TC-7 cells. We concluded that LS180 cells were a suitable model to study MDR1 and CYP3A4 induction, but for drug transport studies Caco-2 parental and TC-7 cells would be preferred as the more physiological model. PMID- 12625869 TI - Efficacy of repifermin (keratinocyte growth factor-2) against abnormalities in gastrointestinal mucosal transport in a murine model of colitis. AB - Human keratinocyte growth factor-2 (KGF-2) is a member of the fibroblast growth factor family that promotes healing of experimental small intestinal ulceration and colitis. The aim of this study was to determine whether repifermin, a truncated form of recombinant human KGF-2, reverses abnormalities in colonic mucosal transport in a murine model of dextran sulfate sodium (DSS)-induced colitis. Male Swiss-Webster mice were given 4% DSS in drinking water for 7 days and then normal drinking water for 3 days. Repifermin (5 mg kg(-1), i.p.) or vehicle was administered daily for 7 days starting on Day 4 of DSS exposure. On Day 10, net ion transport was measured electrophysiologically in colonic mucosal sheets. Repifermin significantly reduced DSS-induced colonic inflammation measured by tissue myeloperoxidase activity. Concurrently, in colonic tissue taken from mice treated with repifermin, there was a normalization of basal potential difference and short circuit current, and an improvement in the secretory responses to stimulation of muscarinic and ganglionic cholinoceptors. In control mice, repifermin did not interact directly with colonic epithelial cells or intramural neurones to induce immediate changes in net electrogenic transport. The results suggest that repifermin therapy may improve the mucosal electrogenic transport that is impaired during colitis. PMID- 12625870 TI - Age-related alteration of haloperidol-serum protein binding. AB - Serum haloperidol levels were determined in 59 patients, 50-88-years-old, with psychosis, receiving long-term treatment with haloperidol. Although the total (bound and free form) haloperidol level in serum showed a linear correlation with daily dose, there was a larger variation in the relationship between free form and the daily dose compared with total because of inter-individual variation in the serum protein binding of haloperidol. The free fraction of haloperidol in serum increased with age. There was no difference in the ratio of total haloperidol level per daily dose between the adult and elderly groups, whereas the ratio of free haloperidol level per daily dose was significantly higher in the elderly than in the adult group. In the elderly, therefore, the therapeutic window of haloperidol should be assessed using free form level rather than total level, which is influenced by serum protein binding of the drug. PMID- 12625871 TI - Synthesis and biochemical evaluation of some novel benzoic acid based esters as potential inhibitors of oestrone sulphatase. AB - Oestrone sulphatase is an important target in the fight against hormone-dependent breast cancer. In an effort to investigate the reported definitive pharmacophore for oestrone sulphatase and continue our search for potent inhibitors of this enzyme, we have undertaken extensive synthesis, biochemical evaluation and physicochemical property determination of a range of benzoic acid based esters. Here, we report the initial results of our study into a series of straight chain alkyl esters of 4-sulphonylbenzoic acid. Using these compounds, we have investigated the involvement of two physicochemical properties, namely logP and pK(a). The results of this study show that there was a strong correlation between the inhibitory activity and the logP of the parent compound. Within the series of compounds studied, hydrophobicity appears to be a more important factor than pK(a) in determining the overall inhibitory activity. In a previous report, we showed that pK(a) plays an important role in stabilizing the phenoxide ion resulting from the hydrolysis of the sulphamate group. Here, we propose that although pK(a) is an important factor in determining the overall inhibitory activity when a wide range of compounds are considered, both hydrophobicity and pK(a) need to be considered in the design of potential inhibitors of oestrone sulphatase. PMID- 12625872 TI - Binding and functional affinity of some newly synthesized phenethylamine and phenoxypropanolamine derivatives for their agonistic activity at recombinant human beta3-adrenoceptor. AB - Beta(3)-adrenoceptor is the predominant beta-adrenoceptor in adipocytes and has drawn much attention during the investigation for anti-obesity and antidiabetes therapeutics. Thirteen new compounds have been evaluated for their potencies and efficacies as beta(3)-adrenoceptor agonists on human beta(3)-adrenoceptor expressed in COS-7 and Chinese hamster ovary (CHO) cells using radioligand binding assay and cyclic AMP (cAMP) accumulation assay. Phenoxypropanolamine derivatives, SWR-0334NA (([E)-[4-[5-[(3-phenoxy-2-hydroxypropyl)amino]-2-pentene 3-yl] phenoxy]acetic acid sodium salt), SWR-0335SA ((E)-[4-[5-[(3-phenoxy-2 hydroxypropyl)amino]-2-pentene-3-yl] phenoxy] acetic acid ethanedioic acid), SWR 0342SA (S-(Z)-[4-[[1-[2-[(2-hydroxy-3-phenoxypropyl)]amino] ethyl]-1 propenyl]phenoxy] acetic acid ethanedioic acid), SWR-0348SA-SITA ((E)-[4-[5-[(3 phenoxy-2-hydroxypropyl)amino]-2-hexene-3-yl] phenoxy]acetic acid ethanedioic acid) and SWR-0361SA ((E)-N-methyl[4-[5-[(3-phenoxy-2-hydroxypropyl)amino]-2 pentene-3-yl]phenoxy]acetoamide ethanedioic acid) showed higher agonistic activity for the beta(3)-adrenoceptor. Among the compounds tested, SWR0334NA exhibited full agonist activity (%E(max) = 100.26) despite its lower binding affinity (pK(I) = 6.11). Compounds SWR-0338SA ((E)-[4-[5-[(2-phenyl-2 hydroxyethyl)amino]-2-pentene-3-yl] phenoxy]acetic acid ethanedioic acid), SWR 0339SA (S-(E)-[4-[5-[(3-phenoxy-2-hydroxypropyl)amino]-2-pentene-3-yl] phenoxy] acetic acid ethanedioic acid), SWR-0345HA ((E)-2-methyl-3-[4-[2-(2-phenyl-2 hydroxyethyl-amino)ethoxy] phenyl]-2-propenoic acid ethyl ester hydrochloride), SWR-0358SA ((E)-(2-methoxyethyl)-[4-[5-[(3-phenoxy-2-hydroxypropyl) amino]-2 pentene-3-yl]phenoxy]acetoamide ethanedioic acid) and SWR-0362SA ((E)-1-[[[4-[5 [(3-phenoxy-2-hydroxypropyl)amino]-2-pentene-3 yl]phenoxy]acetyl]carbonyl]piperidine ethanedioic acid) had moderate agonistic activity and were phenethylamine and phenoxypropanolamine derivatives. Compounds SWR-0065HA ([4-[2-[3-[[(3,4-dihydro-4-oxo-[1,2,4]-triazino(4,5-a)indol)-lyl]oxy] 2-hydroxypropylamino]ethoxy]phenyl]acetic acid methyl ester hydrochloride), SWR 0098NA ((E)-[4-[3-[(2-phenyl-2-hydroxyethyl)amino]-1-butenyl] phenoxy]acetic acid sodium salt) and SWR-0302HA ([4-[[4-[2-(3-chlorophenoxy-2-hydroxypropyl)amino]-E 2-butenyl]oxy]phenoxy]acetic acid hydrochloride) had very low binding affinity towards beta(3)-adrenoceptors and they did not induce cAMP accumulation. We concluded that compounds SWR-0334NA, SWR-0335SA, SWR-0342SA, SWR-0348SA-SITA and SWR-0361SA were potential agonists of human beta(3)-adrenoceptor. Further investigation on their selectivity towards beta(3)-adrenoceptor could be useful for the exploration of the physiological properties of the beta(3)-adrenoceptor. PMID- 12625873 TI - Iso-S-petasin, a hypotensive sesquiterpene from Petasites formosanus, depresses cardiac contraction and intracellular Ca2+ transients in adult rat ventricular myocytes. AB - Petasites formosanus is an indigenous species of the medicinal plant Petasites which has been used to treat hypertension. Both S-petasin and its isoform iso-S petasin have been shown to be the effective ingredients in P. formosanus. However, their effect on heart function has not been revealed. This study was to examine the effect of iso-S-petasin on cardiac contractile function at the myocyte level. Ventricular myocytes were isolated from adult rat hearts and were stimulated to contract at 0.5 Hz under 1.0 mM extracellular Ca(2+). Contractile properties were evaluated using an IonOptix MyoCam system including peak shortening (PS), time to PS (TPS), time to 90% re-lengthening (TR(90)) and maximal velocity of shortening/re-lengthening (+/-dL/dt). Intracellular Ca(2+) properties were assessed by fura-2 and presented as Ca(2+)-induced Ca(2+) release (CICR) and intracellular Ca(2+) decay. Acute application of iso-S-petasin (10(-7) to 10(-4) M) elicited a concentration-dependent inhibition in PS and CICR, with maximal inhibitions of 51.0% and 31.0%, respectively. Iso-S-petasin also induced a concentration-dependent inhibition of+/-dL/dt without affecting TPS, TR(90), baseline intracellular Ca(2+) level or intracellular Ca(2+) decay. Elevation of extracellular Ca(2+) from 1.0 mM to 2.7 mM significantly antagonized the iso-S petasin-induced depression in PS and CICR. These results demonstrated a direct depressant action of iso-S-petasin on ventricular contraction, which may work in concert with its antihypertensive action to reduce the cardiac load. The iso-S petasin-induced decrease in CICR may play a role in its cardiac depressant effect. PMID- 12625874 TI - Protective effects of benidipine on hydrogen peroxide-induced injury in rat isolated hearts. AB - We investigated the effects of benidipine (hydrochloride), a calcium antagonist, on hydrogen peroxide (H(2)O(2))-induced injury in Langendorff-perfused rat hearts. The hearts were aerobically perfused at a constant flow and exposed to H(2)O(2) (600 micromol L(-1)) for 4 min, resulting in the oxidative stress induced myocardial dysfunction (e.g., decrease in the left ventricular developed pressure) and myocardial cell injury (e.g., increase in the release of lactate dehydrogenase). Pretreatment of the hearts with benidipine or nifedipine was performed for 20 min until the start of H(2)O(2) exposure. Benidipine at 1 nmol L(-1) and nifedipine at 10 nmol L(-1) decreased the myocardial contractility and perfusion pressure to a similar degree in the hearts under normal conditions. Benidipine (1 nmol L(-1)) significantly reduced the H(2)O(2)-induced myocardial damage. Nifedipine (10 nmol L(-1)) also tended to exhibit similar effects. Benidipine inhibited the increase in tissue lipid peroxidation induced by H(2)O(2). The results suggest that, in addition to the calcium antagonism, benidipine possesses other actions responsible for the cardioprotective effects, to which the antioxidant activity of benidipine may partly contribute. PMID- 12625876 TI - Behavioural evidence of agonist-like effect of isoteoline at 5-HT1B serotonergic receptors in mice. AB - Isoteoline is a compound of aporphine structure derived from the alkaloid glaucine. Previous studies with isoteoline have shown antagonistic activity at 5 HT(2C) serotonergic receptors. We have investigated whether isoteoline interacts with 5-HT(1B) receptors. An isolation-induced social behavioural deficit test in mice was used as a model of stimulation of these receptors. The deficit in the behaviour of isolated mice in this experimental procedure was reported to be sensitive to 5-HT(1B)-receptor stimulation, since agonists at these receptors are capable of reversing it. In our study, we used N-(3 trifluoromethylphenyl)piperazine (TFMPP) (2 mg kg(-1)) as a reference agonist at these receptor sites. TFMPP completely restored the normal behaviour of the isolated mice. Its effect was prevented by propranolol (4 mg kg(-1)), a beta adrenergic receptor antagonist with a high affinity for 5-HT(1B) receptors, which was inactive by itself. When isoteoline was given before TFMPP, it did not prevent the effect of the latter. Given alone at doses of 0.25, 1, 4 or 8 mg kg( 1), isoteoline showed an effect of its own to normalize the behaviour of isolated mice. The effect of isoteoline (1 mg kg(-1), i.p.) was antagonized by pretreatment with propranolol, indicating that it was mediated through stimulation of 5-HT(1B) receptors. Repeated treatment with isoteoline (1 mg kg( 1), 2 x 3 days, i.p.) produced tolerance to its effect and significantly attenuated the effect of TFMPP, when animals were tested 16 h after the last injection. In conclusion, the results provided functional evidence of agonist like activity of isoteoline at the 5-HT(1B) receptors. PMID- 12625875 TI - Involvement of nuclear factor-kappaB in lipoteichoic acid-induced cyclooxygenase 2 expression in RAW 264.7 macrophages. AB - We have investigated the role of protein kinase C (PKC) and nuclear factor-kappaB (NF-kappaB) in cyclooxygenase-2 (COX-2) expression caused by Staphylococcus aureus lipoteichoic acid in RAW 264.7 macrophages. A phosphatidylcholine phospholipase C (PC-PLC) inhibitor (D-609) and a phosphatidylinositol phospholipase C (PI-PLC) inhibitor (U-73122) attenuated lipoteichoic acid-induced COX-2 expression, while a phosphatidate phosphohydrolase inhibitor (propranolol) had no effect. Two PKC inhibitors (Go 6976 and Ro 31-8220) and the NF-kappaB inhibitor, pyrrolidine dithiocarbamate (PDTC), also attenuated lipoteichoic acid induced COX-2 expression. Lipoteichoic acid resulted in a decrease in PKC activity in the cytosol and an increase in PKC activity in membranes. The lipoteichoic acid-induced translocation of p65 NF-kappaB from the cytosol to the nucleus was inhibited by D-609, U-73122, Go 6976, Ro 31-8220, and PDTC, but not by propranolol. The results suggested that lipoteichoic acid might have activated PC-PLC and PI-PLC to induce PKC activation, which in turn initiated NF-kappaB activation, and finally induced COX-2 expression in RAW 264.7 macrophages. PMID- 12625877 TI - Anti- and pro-oxidant activity of rutin and quercetin derivatives. AB - Some semi-synthetic flavonoids, particularly derivatives of rutin, are used as therapeutic agents in the treatment of diseases involving free radicals. Here, for the first time, a complete study has been made of the relationship between the structure of such molecules and their superoxide, hydroxyl and peroxyl radical scavenging activity. The molecules chosen for this study were rutin, its aglycone (quercetin), and their methyl ethyl and hydroxyl-ethyl derivatives. Our results are consistent with the general agreement on the structural requirements for free radical scavenging activity. Moreover, we have shown that alkylation of the hydroxyl in position 7 enhanced the scavenging, and also that in a Fenton reaction system, some quercetin derivatives with free catechol moiety or free hydroxyl in position 3 (or both) were pro-oxidant, through superoxide radical and hydrogen peroxide production. Although the structural features needed for pro oxidant activity are not entirely clear, it appears that to avoid pro-oxidant behaviour, the hydroxyl group in position 3 should be blocked to prevent its auto oxidation. Thus, flavonoids cannot only be considered purely as antioxidants, since under certain reaction conditions they can also display pro-oxidant activity. This unexpected behaviour could explain, in part, the observed toxicity of some flavonoids in-vivo. PMID- 12625878 TI - Depression of phenytoin metabolic capacity by 5-fluorouracil and doxifluridine in rats. AB - It has been found in clinical practice that the serum level of phenytoin, of which metabolism is mediated by hepatic CYP2C enzymes, was markedly elevated by co-administration of 5-fluorouracil (5-FU) and doxifluridine (5'-deoxy-5 fluorouridine; 5'-DFUR), a prodrug of 5-FU, but the detailed mechanisms are unclear. A study using rats was undertaken to examine the effects of 5-FU and 5' DFUR on phenytoin metabolism in hepatic microsomes and phenytoin pharmacokinetics in-vivo. Neither 5-FU nor 5'-DFUR exhibited direct inhibitory effects on hepatic microsomal phenytoin p-hydroxylation, a major metabolic route catalysed by CYP2C in rats, as in humans. 5-FU and 5'-DFUR were injected intraperitoneally into male rats as single doses (1.68 mmol kg(-1)) and repeated doses (0.24 mmol kg(-1) for 7 days). Control rats received vehicle alone. A significant reduction in the activity of phenytoin p-hydroxylation was observed 4 days after the last administration irrespective of the agents and their treatment regimens, although the activity was unchanged on Day 1. Pharmacokinetic analysis of phenytoin revealed that the elimination rate constant and the total clearance was decreased by 70-75% in both the 5'-DFUR-treated and 5-FU-treated rats, indicating that the decrease in the metabolic capacity of phenytoin was responsible for the change in phenytoin disposition in-vivo. On the other hand, 5-FU significantly depressed the total P450 content, NADPH cytochrome c reductase activity and activities of progesterone hydroxylations. However, the depressive effects of 5'-DFUR were not very potent relative to those of 5-FU, which can be explained by the fact that 5 FU is derived from 5'-DFUR to only a small extent. According to a recent report, phenytoin p-hydroxylation and progesterone 2alpha-/21-hydroxylations share common CYP2C enzymes as their catalysts. Because there was a difference in the modulation profiles between phenytoin p-hydroxylation and progesterone 2alpha-/21 hydroxylations after exposure to 5'-DFUR, 5'-DFUR might modulate phenytoin metabolism without loss of catalytic ability for other substrates, unlike 5-FU. The present study suggested that the down-regulation of hepatic CYP2C enzymes occurs by 5-FU exposure even at a low level, and provided a fundamental explanation for the drug interaction encountered in clinical practice. PMID- 12625879 TI - Iatrogenic perforation in endoscopic stapling diverticulotomy for pharyngeal pouches. AB - In pharyngeal pouch surgery, the relatively new technique of endoscopic stapling diverticulotomy has a number of advantages over more traditional surgical treatments, such as Dohlman's procedure and open pouch excision, and now seems to be the procedure of choice. However, a number of iatrogenic perforations and deaths have been reported with this procedure. We present three cases of iatrogenic perforations occurring during endoscopic stapling of a pharyngeal pouch by different surgeons in our unit, and review the management, causes and prevention of this potentially life-threatening complication. PMID- 12625880 TI - Functions of the mastoid cell system: auto-regulation of temperature and gas pressure. AB - This article presents a new approach to understanding the physiological functions of the mastoid cell system. It is suggested that the cell system, in combination with the continuous blood flow through the adjacent large vessels, makes up a compound functional unit that serves to protect the sensitive vestibular part of the inner ear from inadequate stimulation by external temperature changes. By virtue of the large surface area of the cell system mucosa with respect to the enclosed gas volume, the mastoid cell system may also work as a pressure regulator. Variations of the bi-directional exchange of fluid over the capillary network in the mucosa will change the size of the lumen that is available for the gas in the cell system. Volumes of gas and fluid can thus be exchanged to keep the intratympanic pressure within physiological limits. The process is most effective in a cell system with a high area-to-volume ratio. PMID- 12625882 TI - Current practices of cocaine administration by UK otorhinolaryngologists. AB - The aim of this study was to determine which anaesthetic and vasoconstrictor preparations UK Otorhinolaryngologists use for rhinological surgery, with particular reference to cocaine and adrenaline. The incidence and types of adverse reactions to cocaine were also recorded. A postal survey of all BAO-HNS consultant members was performed. Of the 360 consultant surgeons included in the survey, the majority still use peri-operative cocaine on a regular basis, 66 per cent use cocaine and adrenaline together and more than 40 per cent use cocaine in paediatric patients. Sixteen per cent of respondents did not use cocaine. Only 11 per cent of surgeons had experienced cocaine toxicity in their patients, with only one recorded case of mortality. Most surgeons in the UK use cocaine because of the superior operative field it provides and because they consider it to be safe even with adrenaline. The actual incidence of adverse reactions to cocaine is low, with serious complications being less common than the risks from general anaesthesia. Cocaine remains a valuable agent in the armamentarium of the rhinologist. PMID- 12625881 TI - Vestibular-evoked myogenic potential in patients with unilateral vestibular neuritis: abnormal VEMP and its recovery. AB - The incidence of inferior vestibular nerve disorders in patients suffering from unilateral vestibular neuritis and the recovery of these disorders were evaluated by monitoring the vestibular-evoked myogenic potential (VEMP). Eight patients ranged from 21 to 73 years that suffered from unilateral vestibular neuritis underwent VEMP and caloric testing. Abnormal VEMP was observed in two of the eight patients with unilateral vestibular neuritis. Two patients were diagnosed as having an inferior vestibular nerve disorder. One of these patients showed recovery of the inferior vestibular nerve function as assessed by the VEMP. Disorders of the inferior vestibular nerve function and their recovery was confirmed by our current results. The time course of recoveries of the superior and inferior vestibular nerve systems were similar in the two patients. PMID- 12625883 TI - Biomaterials in laryngotracheal surgery: a solvable problem in the near future? AB - Clinical success of laryngotracheal protheses are constrained by a combination of biocompatible response in the host and a suitable functional rehabilitation. This review considers clinical limits of different materials commonly used in ear, nose and throat surgery and will more particularly focus on titanium, one material recognized to be well tolerated in implantology. PMID- 12625884 TI - The prevalence of Helicobacter pylori infection in malignant and premalignant conditions of the head and neck. AB - Helicobacter pylori is an accepted cause of chronic active gastritis and has a major causative role in peptic ulceration. It is a gastric carcinogen. Its role in non-ulcer dyspepsia (NUD) is less clear; yet 50 per cent of patients with NUD are infected with H pylori. H pylori has been investigated in several other organ systems, but has not been investigated extensively in squamous cell carcinoma of the upper aerodigestive tract, a region which could be directly exposed to the bacterium by gastro-oesophageal reflux (GOR). In this study 61 patients with severe laryngeal dysplasia or frank carcinoma of the head and neck are striated by age, investigated for the presence of antibodies to H pylori and compared to age and sex matched controls. In the age group of 46-61 years, the presence of H pylori antibodies was marginally greater in the experimental (63.0 per cent) than the control group (40.7 per cent) (Pearson Chi square p = 0.055, Fisher 2-sided exact test p = 0.066). When combining this age group with the younger age group and thereby creating two roughly equal groups (n = 31 and n = 30) there was also a statistical trend towards increased positivity in the experimental group. These findings are discussed in the light of other studies with gastro-oesophageal reflux disease (GORD). PMID- 12625885 TI - Microbial contamination of non-disposable instruments in otolaryngology out patients. AB - Nosocomial infections are an important cause of morbidity and contaminated equipment may contribute to this. There has been little work concerning contamination of non-disposable equipment in Otolaryngology. This study investigates the efficacy of the cleaning regimen for non-disposable instruments in an Otolaryngology out-patients clinic. Instruments were swabbed before each clinic, when they had been autoclaved, then after use on patients, when they had been washed with detergent and chlorhexidine. Swabs of 86 instruments were cultured using standardized microbiological techniques. Fifteen instruments (17 per cent) were contaminated, most with coagulase negative staphylococci. Two specimens of Micrococcus luteus were cultured and one each of Pseudomonas aeruginosa, Acinetobacter lwoffii and Aureobacterium spp. Micrococcus luteus and coagulase negative staphylococci may represent skin contaminants, but Aureobacterium spp. and Acinetobacter lwoffii can be sources of nosocomial infection. Pseudomonas aeruginosa is a potentially serious pathogen and is implicated in the aetiology of otitis externa. These findings question the efficacy of the current cleaning techniques. PMID- 12625886 TI - Does receiving a copy of correspondence improve patients' satisfaction with their out-patient consultation? AB - It is standard practice to write to a patient's general practitioner (GP) following an out-patients consultation. This study set out to assess whether sending a copy of this letter to the patient improves their satisfaction with the consultation. Two hundred patients were randomly assigned to receive or not to receive a copy of their GP letter. Their satisfaction was then assessed by means of a postal questionnaire. The two groups were compared to ensure that their was no significant difference between them with regard to any other aspect of their consultation. Those who did not receive a copy letter had a median overall satisfaction score of 7.75 whilst those who did had a median score of 9.0 (p = 0.014). The only other factors predictive of overall satisfaction were receiving an explanation of the problem and spending sufficient time with the doctor. Sending patients a copy of correspondence to their GP is one means of aiding communication and improving overall satisfaction. PMID- 12625887 TI - Ipswich lacrimal tube: pedicle nasal septal tube for the reconstruction of lacrimal drainage passage. AB - Canalicular blockage of the lacrimal system still remains a major challenge for oculoplastic surgeons. The conventional treatment of conjunctival dacryocystorhinostomy using the Lester Jones bypass tube is often associated with tube migration and foreign body reaction. The authors report a new technique to reconstruct the lacrimal passage on a single patient with severe canalicular obstruction following repeated failures from previous Lester Jones tube operations. An epithelial-lined tube was recreated between the nasal cavity and the conjunctiva using a superiorly based mucoperichondrial flap from the nasal septum (Ipswich lacrimal tube). The pedicle of the flap was divided six weeks following the operation. The patient was still symptom free two years following the operation together with a positive dye test confirming the patency of the new lacrimal tube. PMID- 12625888 TI - Endoscopic assisted external approach anterior ethmoidal artery ligation for the management of epistaxis. AB - Anterior ethmoidal artery ligation is a well-established surgical procedure in the management of epistaxis. We describe a procedure of anterior ethmoidal artery ligation via minimal access external surgery with the use of a rigid endoscope. This is, as far as we are aware, the first description of an external approach endoscopic anterior ethmoidal artery ligation. PMID- 12625889 TI - Carbon monoxide poisoning and sensorineural hearing loss. AB - Exposure to carbon monoxide is a well-recognized cause of morbidity and mortality. Both acute accidental poisoning and chronic exposure are associated with a range of adverse health effects. We report two cases of carbon monoxide poisoning with the associated phenomenon of sensorineural hearing loss. Although hearing loss as a result of acute carbon monoxide exposure has previously been described, here we emphasize the need to consider chronic exposure to carbon monoxide as a potentially reversible cause of sensorineural hearing loss if diagnosed and treated early. PMID- 12625890 TI - Permanent bilateral acoustic trauma due to air bag deployment in a young female adult. AB - Air bag safety systems have significantly reduced the number of occupant injuries from road traffic accidents (RTA). However air bag deployment is also associated with unavoidable risks. We report the acoustic trauma incurred by a young female driver who was a heavy smoker as a consequence of air-bag deployment in a low speed RTA and the sparing of her child seated in the rear. PMID- 12625891 TI - Surgical emphysema following nose blowing. AB - A 20-year-old woman developed gross surgical emphysema tracking down to the chest after an episode of nose blowing. Fortunately there were no ocular complications and the patient settled with conservative management. Follow-up showed no recurrence. The case presented illustrates the very rare complication of orbital fracture following nose blowing. PMID- 12625892 TI - Delayed diagnosis of laryngeal foreign body. AB - Aspiration of a foreign body is a recognized cause of accidental death in children. Paediatricians are aware of the symptoms of inhaled foreign bodies in the lower respiratory tract. However, symptoms which suggest impaction in the larynx do not appear to raise the same index of suspicion of a foreign body. One case of laryngeal foreign body is described with a delay in diagnosis of five days. The clinical presentation, investigations and management are discussed. PMID- 12625893 TI - An unusual pattern of intubation injury? AB - Post-intubation laryngeal dysfunction is well documented. Both common and rare patterns of injury have been described in the literature. An unusual pattern of intubation injury is described in this case report. The different patterns of post-intubation laryngeal injury are discussed as well as the possible aetiology in the case described. PMID- 12625894 TI - A rare case of Crohn's disease in head and neck surgery. AB - Oral manifestations of Crohn's disease may present to the Otolaryngologist in the form of ulceration, glossitis and odynophagia, but rarer examples of presentation are also recognized. In this case a 24-year-old lady presented with cervical lymphadenopathy, the subsequent investigation of which resulted in the identification of the disease both in this node and in the tonsils. It is noted that these lesions may precede the classical intestinal manifestations and so the Otolaryngologist could aid in the initial diagnosis of the disease. The pathogenic possibilities of how Crohn's disease might have arisen in these and other extra-intestinal regions, are discussed. PMID- 12625895 TI - Bilateral parotid and submandibular gland enlargement: rare features of Wegener's granulomatosis. AB - Wegener's granulomatosis is a potentially fatal disease of unknown origin affecting mainly the upper and lower respiratory tracts and kidneys. Prompt recognition of the more unusual presentations of the disease is necessary to ensure early treatment. We present a case of a 46-year-old female with bilateral submandibular and parotid gland enlargement. Parotid or submandibular salivary gland enlargement is a rare presenting feature of Wegener's granulomatosis. Common to 80 per cent of these cases is nasal involvement, while ear pathology or lung lesions may occur in the remaining cases. The diagnosis is both clinical and pathological, biopsy of suspicious tissue, serum c-ANCA levels and a chest X-ray are valuable investigations. Treatment with immunosuppressive therapy is essential, and usually ensures a long-term remission. PMID- 12625896 TI - Bilateral parotid cysts as presentation of Sjogren's syndrome. AB - We present a case of a 40-year woman with bilateral parotid salivary gland enlargement as presentation of primary Sjogren's syndrome. Computed tomography (CT) and magnetic resonance imaging (MRI) showed parotid cysts, suggestive of cystic benign lymphoepithelial lesions. A sub-labial biopsy confirmed the syndrome. After 24-month follow-up, the left parotid cysts remain the same, whereas other cysts have appeared in the right parotid gland. Parotid involvement in Sjogren's syndrome is discussed. PMID- 12625916 TI - Development and standardization of a rapid, PCR-based method for the detection of Wuchereria bancrofti in mosquitoes, for xenomonitoring the human prevalence of bancroftian filariasis. AB - PCR has recently been studied as a promising tool for monitoring the progress of efforts to eliminate lymphatic filariasis. PCR can be used to test concurrently at least 30 pools, with as many as 40 mosquitoes in each pool, for the presence of filarial larvae. The SspI PCR assay for the detection of Wuchereria bancrofti DNA in pools of mosquitoes has been used since 1994 in a variety of laboratories worldwide. During that time, the original assay has been modified in these different laboratories and no standardized assay currently exists. In an effort to standardize and improve the assay, a meeting was held on 15-16 November 2001, at Emory University in Atlanta, with representatives from most of the laboratories currently using the assay. The first round of testing was designed to test the four most promising methods for DNA extraction from pools of mosquitoes. Two of the four methods stood out as clearly the best and these will be now optimised and evaluated in two further rounds of testing. PMID- 12625917 TI - Xenomonitoring of Culex quinquefasciatus mosquitoes as a guide for detecting the presence or absence of lymphatic filariasis: a preliminary protocol for mosquito sampling. AB - A protocol for the collection of resting, blood-engorged Culex quinquefasciatus Say and their examination for microfilariae has been developed as a way of detecting whether lymphatic filariasis (LF) occurs in a particular locality. The protocol was first implemented in a pilot study in Trinidad, West Indies. For gathering prevalence data, such xenomonitoring is a suitable alternative to the use of human bait, which is ethically questionable. The resting mosquitoes were collected, either indoors or outdoors, using electrical and mouth aspirators. A 'cocoeya broom', made from a bunch of the midribs of coconut-palm leaves, was found to be useful in flushing out the mosquitoes resting in hard-to-reach areas within bedrooms. The rationale behind the strategy and the five-step methodology, of householder notification, mapping, preparation of equipment, mosquito collecting and laboratory processing, are described. Data from the pilot study indicate that this xenomonitoring protocol may be applicable worldwide, albeit with modifications to take account of variations in the vector species involved and their ecology and resting behaviour. PMID- 12625918 TI - Field application of PCR-based assays for monitoring Wuchereria bancrofti infection in Africa. AB - Approximately 50 million people in Egypt and sub-Saharan Africa have bancroftian filariasis and together they represent about a third of all cases of lymphatic filariasis (LF) world-wide. Currently, the Global Programme to Eliminate Lymphatic Filariasis, which was launched by the World Health Organization (WHO) in 1998, is largely based on repeated annual cycles of mass drug administration (MDA) to endemic populations. Also, some countries, including Egypt, are taking steps to improve LF vector-control interventions, to break the transmission cycle more effectively than is achievable with MDA alone. New tools and strategies for monitoring and evaluating elimination campaigns are needed. The last 20 years have witnessed dramatic advances in the diagnosis of LF for epidemiological purposes. The recent introduction and development of molecular technologies have moved parasite-detection systems from traditional methods (that are labour intensive, tedious and often impractical) to improved PCR-based assays that have considerable potential for field use. The present article highlights the strengths and limitations of the PCR-based assays when used to detect filarial infections in mosquitoes (particularly for the xenomonitoring of elimination campaigns). PMID- 12625919 TI - Progress towards, and challenges for, the elimination of filariasis from Pacific island communities. AB - The Pacific Programme for the Elimination of Lymphatic Filariasis (PacELF) - the first regional campaign to attempt to eliminate filariasis as a public-health problem - is using five, annual, mass drug administrations (MDA) of diethylcarbamazine (DEC) plus albendazole to stop transmission. In 2001, nine countries and territories covered by the programme had begun annual MDA campaigns, with population treatment coverages ranging from 52% to 95%. By the end of 2002, it is anticipated that 11 countries/territories will have begun such MDA campaigns. Even with high MDA coverage, the efficiency of Aedes polynesiensis as a vector of Wuchereria bancrofti may limit the effectiveness of the elimination campaigns in some countries. In areas of limited MDA coverage, additional strategies, such as vector control (as a adjunct to the MDA), or alternative approaches, such as the use of DEC-fortified salt, may be necessary to stop transmission. PMID- 12625920 TI - Operational issues in the control of the vectors of Brugia. AB - An estimated 13 million people in the Oriental Region have brugian filariasis. The filarial parasites that cause this disease exist in periodic and sub-periodic forms and are transmitted by four genera of mosquito: Anopheles, Mansonia and, less frequently, Coquillettidia and Ochlerotatus. In most endemic countries, control of the disease has been entirely based on chemotherapy, although house spraying and use of insecticide-treated bednets can be quite effective against the vectors of nocturnally periodic Brugia malayi and B. timori. The vector control methods that may be applied against the Mansonia mosquitoes that transmit the parasites causing sub-periodic brugian filariasis are reviewed here. Most of the conventional methods for controlling the immature, aquatic stages of mosquitoes have proved unsatisfactory against Mansonia. The reason is that, unlike the those of other genera, the larvae and pupae of Mansonia spp. are relatively immobile and obtain air not at the water surface but from the underwater roots, stems and leaves of floating plants to which the larvae and pupae attach. Removal of host plants can be very effective in reducing Mansonia productivity, whereas large-scale use of herbicides is restricted by the potential adverse effects on the ecosystem. Environmental management in water development projects remains the best option. PMID- 12625921 TI - Cost-effectiveness of the use of vector control and mass drug administration, separately or in combination, against lymphatic filariasis. AB - The costs and effects of two intervention strategies for the control of bancroftian filariasis-annual mass drug administrations (MDA) with a combination of diethylcarbamazine and ivermectin, with or without integrated vector control (VC)-were estimated in rural villages in South India. The aim was to compare the cost-effectiveness of MDA alone with that of MDA plus VC. Control of the local vector, Culex quinquefasciatus, was based on the application of polystyrene beads to cesspits, the treatment of drains with larvicidal Bacillus sphaericus and the stocking of wells with larvivorous fish. An itemized cost menu was used to cost MDA and MDA + VC, retrospectively. The annual transmission potential was used to assess the direct outcome of the disease-control methods, whereas the prevalence and intensity of microfilaraemia were used as indicators of the impact of each method. The per-capita costs were 1.49 U.S. dollars for two rounds of MDA, 1.70 U.S. dollars for 2 years of VC and, therefore, 3.19 U.S. dollars for 2 years of MDA + VC. Integration of VC with MDA did not appear to be cost-effective: it cost an estimated 1.80 U.S. dollars to stop an infective mosquito biting a villager using MDA alone but 3.32 U.S. dollars to achieve the same result using MDA + VC. Similarly, the cost to reduce the prevalence of microfilaraemia in a three village group by 1% was only 96.62 U.S. dollars for MDA alone but 201.16 U.S. dollars when vector control was integrated. The implications of these results for the control and elimination of filariasis in Indian village communities, and the options for sharing and minimizing costs, are discussed. PMID- 12625922 TI - Impact of insecticide-treated materials on filaria transmission by the various species of vector mosquito in Africa. AB - Nocturnally periodic bancroftian filariasis is maintained by three mainly endophilic vectors in East Africa: Culex quinquefasciatus, Anopheles funestus and the An. gambiae complex. Permethrin-impregnated bednets provide considerable protection against these mosquitoes, but the species respond differently. The degree of protection conferred by treated bednets was determined in Kenyan communities where all three vectors actively transmit Wuchereria bancrofti. The annual transmission potential in the communities (i.e. an estimate of the number of human infective, third-stage larvae of W. bancrofti inoculated into each villager each year) was reduced by 92%, through the nets' impact on vector biting rates (reduced by 22%) and their cumulative impact on the annual infective biting rate (reduced by 95%). Thus a modest reduction in the numbers of mosquitoes biting humans, attributable to the use of the insecticide-treated nets, strongly suppressed the risk of W. bancrofti transmission. PMID- 12625923 TI - Use of floating layers of polystyrene beads to control populations of the filaria vector Culex quinquefasciatus. AB - Floating layers of polystyrene beads suffocate mosquito larvae and pupae and inhibit egg laying. The layers are very durable in breeding sites with water contained within walls, as in wet pit latrines and soakage pits. In some areas such pits constitute an important breeding site for Culex quinquefasciatus. Trials have been conducted in communities in Zanzibar, Tanzania, and in Tamil Nadu, India, where such mosquito populations were the vectors of Wuchereria bancrofti. In each case, treatment of all the pits with polystyrene beads was integrated with mass treatment of the people with antifilarial drugs-in Zanzibar in 1988 with diethylcarbamazine (DEC) and in India in the 1990s with DEC plus ivermectin. The results were compared with those in communities with the mass drug treatment alone and with control communities with neither treatment. The polystyrene-bead treatments greatly and sustainably reduced the vector populations. Comparison of the communities after drug treatment ceased showed that this form of vector control contributed markedly to the prevention of a resurgence of filarial infection. Where Cx. quinquefasciatus breeding in pits form a major component of the vector population, use of polystyrene-bead layers could assist considerably in the process of eliminating lymphatic filariasis by mass drug administration. PMID- 12625924 TI - Control of the Aedes vectors of the dengue viruses and Wuchereria bancrofti: the French Polynesian experience. AB - In most of the 130 islands of French Polynesia, the stenotopic mosquitoes Aedes aegypti (the main local vector for the viruses causing dengue) and Aedes polynesiensis (the main local vector of Wuchereria bancrofti) share many breeding sites in water containers such as discarded cans, coconut shells, buckets and water-storage pots and drums. In addition to selective application of insecticides, non-polluting methods of controlling these mosquitoes have been evaluated during the last decade in two main ecological situations: (1) villages, where Aedes breeding sites are typically peridomestic; and (2) flooded burrows of land crabs, the major source of Ae. polynesiensis throughout the South Pacific region. Large-scale trials of biological control agents, such as mosquito fish (Gambusia affinis and Poecilia reticulata) and copepods (Mesocyclops aspericornis), and of integrated-control strategies have demonstrated the efficacy of certain techniques and control agents against the target Aedes populations in some village situations. Generally, mechanical methods (the use of layers of polystyrene beads against mosquito larvae and pupae, and screening against adult mosquitoes) were more efficient than use of the biological control agents. By integrating several methods of control, mosquito densities (as measured by human-bait collections and larval surveys) were reduced significantly compared with the results of concurrent sampling from untreated villages, and control remained effective for months after the interventions ceased. In land crab burrows, the first attempts to control Aedes larvae used bacterial agents (Bacillus thuringiensis) and predatory copepods gave disappointing results. Mesocyclops aspericornis could be an effective control agent if the burrows were constantly flooded, but most burrows dry out and refill periodically, so copepod populations do not survive. As it proved difficult to reach all corners of the long sinuous burrows with any control agent, larvicidal (chlorpyrifos-methyl) baits were developed for foraging crabs to carry into their burrows. This novel technique proved to be effective and could become the method of choice for treating crab burrows. Further research is underway to find the optimum (biological or chemical) larvicidal ingredient for the crab bait. Despite the ecological and logistical challenges of controlling the Aedes vectors of the dengue viruses and W. bancrofti in so many scattered islands, the French Polynesian experience indicates that relatively simple methods can be integrated and applied effectively and economically. Operationally, however, success also depends on a strong political commitment and on at-risk communities that are sufficiently motivated to maintain a good level of Aedes control. PMID- 12625925 TI - Wolbachia as a potential tool for suppressing filarial transmission. AB - There is currently a great deal of interest in Wolbachia because of their wide distribution in arthropods and filarial nematodes and their striking effects on the biology of their hosts, including a possible role in speciation. They manipulate the reproduction of arthropod hosts through various effects on their hosts' biology, particularly cytoplasmic incompatibility (CI), to increase the proportion of infected individuals in the population, often to the point of fixation. This ability of Wolbachia to sweep through host populations indicates several potential applications of Wolbachia in the control of mosquito-borne disease. One uses Wolbachia-induced CI as a form of sterile-insect technique, to suppress mosquito populations. Another envisages the application of CI for population replacement, with the intention of preventing the transmission of human pathogens, by substituting desirable genotypes, including those carried in transgenes. A third possibility is to use Wolbachia to reduce the survival of mosquito populations and thereby reduce their ability to transmit the infection. This article provides an overview of the biological effects of Wolbachia on arthropod hosts, with discussion of the possible future exploitation of these effects in the control of filariasis. PMID- 12625926 TI - Entomological monitoring of annual mass drug administrations for the control or elimination of lymphatic filariasis. AB - Together, the interruption of transmission, through annual mass treatments with antifilarial drugs, and the alleviation of suffering in chronic cases form the principal strategy of the Global Programme to Eliminate Lymphatic Filariasis (GPELF). Entomological monitoring of the annual mass drug administrations is necessary to assess transmission levels and progress towards control or elimination of lymphatic filariasis. However, the traditional procedures for entomological monitoring, such as the dissection of mosquitoes caught manually, either as they come to bite at night or as they rest by day, are cumbersome, cost prohibitive and often not feasible in large-scale control programmes. The collection of mosquitoes in traps and the assessment of their infection status using PCR-based assays offer promise and need to be established as a routine part of control programmes. It is proposed that-to verify the interruption of transmission or determine the prevalence of mosquito infection-1% of the villages/communities in each intervention unit be sampled, mosquitoes be collected from at least 100 households in each study village, and 10 female mosquitoes or all of those collected (whichever is the lower number) from each study house then be checked for infection. PMID- 12625927 TI - Limitation and facilitation in the vectors and other aspects of the dynamics of filarial transmission: the need for vector control against Anopheles-transmitted filariasis. AB - In certain filaria-mosquito combinations, the number of infective, third-stage larvae (L(3)) that develop in a mosquito is not proportional to the number of microfilariae (mff) ingested by that mosquito. As the number of mff ingested increases, the yield of L(3) per microfilaria may either increase (in a process known as 'facilitation') or decrease (in a process known as 'limitation'). Each ingested microfilaria that is successful (in terms of reaching the haemocoel) increases (facilitation) or decreases (limitation) the 'permeability' of the stomach wall for the next microfilaria. Limitation is seen in some culicine mosquitoes, especially the Aedes spp. that transmit Wuchereria bancrofti, which, in consequence, become relatively more efficient as vectors as they ingest fewer mff. This phenomenon makes the interruption of filarial transmission by Aedes spp. particularly difficult. As the survival of anopheline mosquitoes is adversely affected by filarial infection, the use of mass drug administrations (MDA) to reduce the prevalence and intensity of microfilaraemias may increase the mean lifespan of some of the local Anopheles species. If these same species also act as vectors of malarial parasites, effective, drug-based control of W. bancrofti may worsen the problem posed by malaria. Therefore, wherever malaria and bancroftian filariasis are co-endemic and caused by parasites transmitted by the same species of mosquito, MDA should be augmented by interventions (use of bednets or house-spraying) against adult Anopheles. PMID- 12625928 TI - Modelling the epidemiology, transmission and control of lymphatic filariasis. AB - Wuchereria bancrofti transmitted by Culex quinquefasciatus accounts for >90% of the global burden of lymphatic filariasis (LF). Recent advances in diagnostic and control tools and a better epidemiological understanding of the disease have led to hope that LF is eradicable. The World Health Organization has helped a number of member countries to launch nation-wide programmes of mass treatment with antifilarial drugs such as diethylcarbamazine, albendazole and ivermectin, for the elimination of this disease. In order to make rational decisions about control strategies, reliable predictions of the long-term impact of such treatment, and of alternative interventions, need to be made, and these can only be based on a sound, quantitative understanding of the population biology of the parasites. Mathematical models have proven valuable in gaining quantitative insights into the population dynamics of the parasites, and may be used to make credible predictions of the likely outcomes of various control strategies. This article provides an overview of the development of the relevant mathematical/statistical models and of their application in studies of the epidemiology, transmission and control of lymphatic filariasis. PMID- 12625929 TI - The genetic diversity of Brazilian Trypanosoma cruzi isolates and the phylogenetic positioning of zymodeme 3, based on the internal transcribed spacer of the ribosomal gene. AB - There is a high level of genetic diversity among the clinical isolates of the taxon Trypanosoma cruzi. The results of recent studies on well defined gene loci have revealed that T. cruzi can be divided into two major phylogenetic lineages, designated T. cruzi I and T. cruzi II. Further intra-lineage polymorphisms were explored, in clinical isolates from five representative regions of Brazil, using the internal transcribed spacers that flank the 5.8S ribosomal RNA gene. These regions were amplified in a PCR and the products were digested with six restriction enzymes before being subjected to RFLP analysis in polyacrylamide gels. The results were used to construct a complex evolutionary tree, in which the isolates were clustered into two major phylogenetic lineages with a low level of similarity. To investigate the evolutionary relationships between the isolates, the PCR products were cloned and sequenced. The results of the subsequent phylogenetic analysis confirmed the presence of the two major groups of T. cruzi and showed that zymodeme 3, a third iso-enzymatic sub-group, is closer to T. cruzi I than to T. cruzi II. PMID- 12625930 TI - The natural history of Sudanese post-kala-azar dermal leishmaniasis: clinical, immunological and prognostic features. AB - In an exploration of the natural history of post-kala-azar dermal leishmaniasis (PKDL), 134 residents of Sudan who had recently been diagnosed as cases of the disease were investigated. In each case, diagnosis had been based on clinical criteria, the temporal relationship between the rash and the treatment of visceral leishmaniasis (VL), positive results in direct agglutination tests (DAT) and/or leishmanin skin tests (LST), and the exclusion of other skin conditions. The mean (S.D.) age of the subjects was 6.4 (3.0) years. Although PKDL appeared commonest among those aged 4-8 years (P < 0.05), it was most severe in children aged 8-O-4-diferulic acid > 8-8-diferulic acid. PMID- 12625947 TI - Phenolic antioxidants: electrochemical behavior and the mechanistic elements underlying their anodic oxidation reaction. AB - Electrochemical analysis has been widely used to assess activities of reductant antioxidants, but the mechanistic factors that determine reducing ability and their corresponding correlations remain to be explored further. In the present paper, the reactivity of a selection of phenolic compounds in anodic oxidation was investigated by cyclic voltammetry (CV). The effects of electron-donation ability, deprotonation equilibrium, stability of radicals and chemical hardness on reducing capacity were studied by computational chemistry methods and multiple linear regression analysis. The results suggested that all these factors made significant contributions to reducing potency although electron-donation ability of the parent molecule plays the most important role in the electrode reaction. With the majority of the compounds examined, the occurrence of multiple electrochemical reactions leading to irreversible anodic oxidation was implied by: (i) the characteristics of the radical cation intermediate; (ii) the propensity of the deprotonation reaction; and (iii) the reactivity of the parent molecule. Upon correlation analysis of oxidation potential and computed physicochemical descriptors, some new light was shed on the mechanism by which phenolic compounds act as antioxidant reductants in free radical reactions. A satisfactory multi-descriptor theoretical QSAR model was derived, which might be of predictive significance in the screening or design of new antioxidants. PMID- 12625946 TI - Scavenging of photogenerated oxidative species by antimuscarinic drugs: atropine and derivatives. AB - The quenching ability of photogenerated oxidative species by some antimuscarinic drugs generically named atropines (e.g. atropine [I] eucatropine [II], homatropine [III] and scopolamine [IV]) have been investigated employing stationary photolysis, polarographic detection of dissolved oxygen, stationary and time-resolved fluorescence spectroscopy, and laser flash photolysis. Using Rose Bengal as a dye sensitiser for singlet molecular oxygen, O(2)((1)Delta(g)), generation, compounds I-IV behave as moderate chemical plus physical quenchers of the oxidative species. Correlation between kinetic and electrochemical data indicates that the process is possibly driven by a charge-transfer interaction. The situation is somewhat more complicated employing the natural pigment riboflavin (Rf) as a sensitiser. Compounds I and II complex Rf ground state, diminishing the quenching ability towards singlet and triplet excited state of the pigment. On the other hand, compounds III and IV effectively quench Rf excited states, protecting the pigment against photodegradation. Under anaerobic conditions, semireduced Rf (Rf(.-)) is formed through quenching of excited triplet Rf. Nevertheless, although Rf(.-) is a well-known generator of the reactive species superoxide radical anion by reductive quenching in the presence of oxygen, the process of O(2)((1)Delta(g)) production prevails over superoxide radical generation, due to the relatively low rate constants for the quenching of triplet Rf by the atropines (in the order of 10(7) M(-1)s(-1) for compounds III and IV) in comparison to the rate constant for the quenching by ground state oxygen, approximately two orders of magnitude higher, yielding O(2)((1)Delta(g)). Compound I is the most promising O(2)((1)Delta(g)) physical scavenger, provided that it exhibits the higher value for the overall quenching rate constant and only 11% of the quenching process leads to its own chemical damage. PMID- 12625949 TI - WTEC panel report on tissue engineering. PMID- 12625950 TI - Articular cartilage bioreactors and bioprocesses. AB - This review summarizes the major approaches for developing articular cartilage, using bioreactors and mechanical stimuli. Cartilage cells live in an environment heavily influenced by mechanical forces. The development of cartilaginous tissue is dependent on the environment that surrounds it, both in vivo and in vitro. Chondrocytes must be cultured in a way that gives them the proper concentration of nutrients and oxygen while removing wastes. A mechanical force must also be applied during the culturing process to produce a phenotypically correct tissue. Four main types of forces are currently used in cartilage-culturing processes: hydrostatic pressure, direct compression, "high"-shear fluid environments, and "low"-shear fluid environments. All these forces have been integrated into culturing devices that serve as bioreactors for articular cartilage. The strengths and weaknesses of each device and stimulus are explored, as is the future of cartilage bioreactors. PMID- 12625952 TI - Serum from patients anesthetized with opiates less effective in the support of chondrocyte growth in vitro. AB - Risk of viral and/or prion disease transmission associated with the use of fetal bovine serum in clinical cell culture has led to the increasing use of autologous human serum in tissue engineering. A relatively large volume of blood is needed and so, to decrease patient discomfort, we have investigated the feasibility of taking blood when the patient is anesthetized. Two serum samples were prepared from each of 22 patients: (1). from the awake patient (PRE) and (2). from the patient 5 min after induction of general anesthesia (PER). The sera were compared for their ability to support the in vitro proliferation of primary human chondrocytes, determined by cell counting. The effects of anesthetic agents on the PER/PRE cell number ratio were established by analysis of variance and stepwise multilinear regression analysis. The PER sample supported higher growth in 2 of 22 patients, equivalent growth in another 11, and significantly lower growth in the remaining 8. Only the opiate analgesics (fentanyl [Sublimaze], alfentanyl [Rapifen], and diamorphine) had a significant and inhibitory effect on chondrocyte proliferation. It is suggested that opiate analgesics be avoided when blood is taken to support the in vitro growth of human cells. PMID- 12625951 TI - Modulation of the contractile and biosynthetic activity of chondrocytes seeded in collagen-glycosaminoglycan matrices. AB - Studies have demonstrated that articular chondrocytes can express the gene for alpha-smooth muscle actin (SMA) and can contract porous polymeric matrices employed for tissue engineering, thereby altering the pore structure and distorting the shape of the scaffold. The objectives of this study were to determine whether an agent known to disrupt microfilament organization in chondrocytes could reduce this contractility and to assess whether there was an association between the contractile behavior of chondrocytes and their biosynthetic activity. Staurosporine, an antibiotic known to inhibit protein kinase C and disrupt cytoskeletal structure, was used as the agent to modulate the chondrocytic phenotype and contractile and biosynthetic activity of serially passaged adult canine chondrocytes seeded in type 1 collagen-glycosaminoglycan scaffolds. Cells in monolayer culture treated with as little as 3 nM staurosporine for 4 days contained type II procollagen, whereas few cells in the untreated control cultures demonstrated type II procollagen synthesis. Treatment with staurosporine also led to a decrease in the amount of SMA synthesized by the cells. Consistent with this decreased expression of the contractile actin isoform, cells cultured in the collagen-glycosaminoglycan scaffolds and treated with 5 nM staurosporine contracted the scaffold significantly less than untreated cells (15% diameter contraction by treated cells, compared with more than 50% contraction by untreated cells). The staurosporine-treated cells were biosynthetically active, displaying higher rates of protein and glycosaminoglycan synthesis, as indicated by rates of incorporation of [(3)H]proline and [(35)S]sulfate, respectively, compared with untreated cells. The long-held notion that changes in cytoskeletal structure influence phenotypic characteristics of cultured chondrocytes may now be extended to relate expression of a specific muscle actin isoform to certain cell processes. Moreover, the finding that chondrocytes with a lower level of expression of SMA and reduced contractility display higher rates of biosynthesis warrants further study. PMID- 12625953 TI - Optimum combination of monolayer and three-dimensional cultures for cartilage like tissue engineering. AB - The autologous chondrocyte transplantation technique has been introduced for the repair of articular cartilage defects. The advantage of transplanting chondrocytes cultured in suspension includes the in vitro expansion of cell numbers. However, the disadvantages include the potential leakage of cells from defects, dedifferentiation of cellular phenotype, and uneven distribution of cells. Transplantation of chondrocytes cultured in collagen gel resolves those problems. However, the expansion of cells in three-dimensional culture is more difficult than in monolayer culture, and for practical reasons only limited numbers of chondrocytes can be obtained from an unloaded area of the knee. To develop a method for the production of high-quality cultured grafts, we investigated the combination of monolayer culture for cell expansion and three dimensional culture for maintenance of cell phenotype. Articular chondrocytes from rabbits were divided into four groups, exposed to various combinations of culture conditions, and cultured for a total of 3 weeks. Each group was evaluated histologically, biochemically, and biomechanically. Our findings showed that the combination of 2 weeks of monolayer culture followed by 1 week of three dimensional culture resulted in the highest chondroitin sulfate levels, sufficient cell numbers, and adequate stiffness of the chondrocyte-collagen composites, giving optimal graft preparation. PMID- 12625954 TI - Functional assessment of tissue-engineered meniscal cartilage by magnetic resonance imaging and spectroscopy. AB - A perfusion bioreactor system was used to grow bioartificial meniscal cartilage tissue in vitro. Magnetic resonance imaging and magnetic resonance spectroscopy methods were used to characterize the flow and perfusion profiles and the growth, distribution, and bioenergetics of the fibrochondrocytes in the resulting constructs. These measurements were correlated with each other and with subsequent histologic analysis. The study has demonstrated that these noninvasive magnetic resonance methods will be useful for designing bioreactor operation strategies and cell scaffolds that lead to the production of tissue-engineered meniscal cartilage constructs with properties resembling those of the native tissue. PMID- 12625955 TI - Characterization of polylactic acid-polyglycolic acid composites for cartilage tissue engineering. AB - The objective of this study was to determine the effects of scaffold composition on the physical properties, adhesion, and growth of bovine articular chondrocytes on polylactic acid (PLA)/polyglycolic acid (PGA) composites. Nonwoven meshes of PGA were coated with PLA, using a solvent evaporation technique that resulted in composites with fractional PLA contents ranging from 0 to 68%. The compressive modulus of scaffolds increased linearly with the addition of PLA, ranging from less than 1 kPa for PGA to approximately 20 kPa for scaffolds with 68% PLA content. The characteristic degradation time of these scaffolds also increased from approximately 5 days for 0% PLA to 45 days for 68% PLA. Addition of PLA decreased cell seeding efficiency from 48% for 0% PLA scaffolds to 27% for 68% PLA scaffolds. Cells seeded onto 27% PLA scaffolds increased 3-fold in number over 4 weeks in culture, whereas cells seeded onto 68% PLA increased only 2-fold in number. Scanning electron microscopy indicated that cells attached to PGA appeared flat with many small processes, whereas those attached to PLA were more rounded. These studies provide important information for the design of scaffolds for cartilage tissue engineering. PMID- 12625956 TI - Poly(D,L-lactic acid)-poly(ethylene glycol)-monomethyl ether diblock copolymers control adhesion and osteoblastic differentiation of marrow stromal cells. AB - Biodegradable polymers, such as poly(lactic acid) (PLA) and poly(lactic coglycolic acid) (PLGA), are attractive materials for tissue engineering because of their degradative and mechanical properties, which permit scaffolds to be tailored to the individual requirements of different tissues. Although these materials support tissue development, their chemical properties offer no control of cell adhesion or function because their surfaces become immediately masked by adsorbing serum proteins when the materials come into contact with body fluids. Furthermore, adhesion proteins undergo conformational changes and a decrease in bioactivity when adsorbed to hydrophobic materials, such as PLA. To overcome these limitations, we modified the properties of PLA by synthesizing a diblock copolymer with poly(ethylene glycol) (PEG), which is known to reduce the amount of adsorbed proteins and to modify their conformation. By altering the PEG content of these diblock copolymers we were able to control the adsorption of adhesion proteins and, because cell adhesion takes place only in the presence of serum proteins, to control cell adhesion and cell shape. Marrow stromal cell differentiation to the osteoblastic phenotype was strongly improved on PEG-PLA compared with PLA, PLGA and tissue culture polystyrene and led to a 2-fold increase in alkaline phosphatase activity and mineralization. PMID- 12625957 TI - Quantitative measures of osteoinductivity of a porous poly(propylene fumarate) bone graft extender. AB - Bioresorbable bone graft substitutes could alleviate disadvantages associated with the use of autografts, allografts, and other synthetic materials. However, little is known about the minimum autograft/extender ratio for a given material at which a sufficient osteoinductive effect is still seen. Therefore, we investigated a bioresorbable bone graft substitute made from the unsaturated polyester poly(propylene fumarate), PPF, at various mixing ratios with autograft. The bone graft extender is cross-linked in the presence of a hydroxylapatite filler and effervescent foaming agents citric acid and sodium bicarbonate. The porous bone graft extender material develops porosity in vivo by generating carbon dioxide during the effervescent reaction, resulting in foam formation and expansion with respective pore sizes of 50 to 1000 microm. In an attempt to determine how much cancellous autograft bone could be extended with the poly(propylene fumarate) material and at which ratio the autograft/extender combination remained supportive of the overall structural integrity of the repairing defect site, we studied the amount of new bone formation on implantation of the materials in 3-mm holes made in the anteromedial tibial metaphysis of Sprague-Dawley rats. The extender formulation was analyzed at high autograft/extender (75% autograft/25% extender) and low autograft/extender (25% autograft/75% extender) mixing ratios and compared with negative (extender alone) and positive (autograft alone) controls. Animals from each of the formulations were killed in groups of eight at 6 weeks postoperatively. Hence, a total of 32 animals were included in this study. Histologic analysis of the healing process revealed enhanced in vivo osteoinduction with the bone graft extender regardless of the autograft loading. Histomorphometry did not show any statistically significant difference between the high and low autograft/extender ratios. All formulations maintained implant integrity and did not provoke sustained inflammatory responses. This study suggested that the presence of even a small amount of autograft within the polymer-based bone graft extender results in significant enhancement of osteoinduction. This finding has immediate applicability to the development of bone graft extender formulations for clinical use. PMID- 12625958 TI - Role of vascular endothelial growth factor in bone marrow stromal cell modulation of endothelial cells. AB - One of the fundamental principles that underlies tissue-engineering strategies using cell transplantation is that a newly formed tissue must acquire and maintain sufficient vascularization in order to support its growth. Enhancing angiogenesis through delivery of growth factors is one approach to establishing a vascular network to these tissues. In this study, we tested the potential of bone marrow stromal cells (BMSCs) to modulate the growth and differentiation activities of blood vessel precursors, endothelial cells (ECs), by their secretion of soluble angiogenic factors. The growth and differentiation of cultured ECs were enhanced in response to exposure to BMSC conditioned medium (CM). Enzyme-linked immunosorbent assays demonstrated that both mouse and human BMSCs secreted significant quantities of vascular endothelial growth factor (VEGF) (2.4-3.1 ng/10(6) cells per day). Furthermore, eliminating the activity of BMSC-secreted VEGF with blocking antibodies completely blocked the CM effects on cultured ECs. These data demonstrate that human BMSCs secrete sufficient quantities of VEGF to enhance survival and differentiation of endothelial cells in vitro, and suggest they may be capable of directly orchestrating angiogenesis in vivo. PMID- 12625959 TI - Application and limitations of chloromethyl-benzamidodialkylcarbocyanine for tracing cells used in bone Tissue engineering. AB - Bone tissue engineering has the potential to provide us with an autologous bone substitute. Despite extensive research to optimize the technique, little is known about the survival and function of the cells after implantation. To monitor the cells, in vivo labeling is the method of choice. In this study we investigated the use of the fluorescent membrane marker chloromethyl benzamidodialkylcarbocyanine (CM-Dil) to label cells used in bone tissue engineering. When applying label concentrations up to 50 microM, cells could be labeled efficiently without negative effects on cell vitality, proliferation, or bone-forming capacity. Porous hydroxyapatite scaffolds were seeded with labeled cells, and up to 6 weeks after implantation in nude mice cells could be traced inside tissue-engineered bone. However, contrary to other reports concerning intramembranous labels, transfer of the label from labeled to unlabeled cells was detected. Transfer occurred both in vitro and in vivo between vital cells and between dead and living cells. To determine when in vivo label transfer happened, devitalized, labeled constructs were implanted for various time periods in nude mice. The presence of vital labeled cells inside these constructs, when evaluated at different implantation periods, indicated transfer of the label. Transfer occurred at 7 days postimplantation when 40 microM label was applied, whereas 10 microM labeled constructs showed transfer 10 days after implantation. These findings indicate that CM-Dil label is useful for in vivo tracing of cells for follow-up periods up to 10 days. This makes the label particularly useful for cell survival studies in tissue-engineered implants. PMID- 12625960 TI - Soft tissue reaction to microgrooved poly-L-lactic acid implants loaded with transforming growth factor beta(3). AB - In both normal and disturbed wound healing, the generation of large, contracting scars can raise serious functional and cosmetic problems. A possible strategy to minimize or avoid the generation of scar tissue surrounding an implant is to apply transforming growth factor-beta(3) (TGF-beta(3)) to the implant. TGF beta(3) (0, 1, or 2.5 microg) was freeze-dried onto poly-L-lactic acid (PLA) microgrooved substrates (width, 10 microm; depth, 1 microm) and implanted subcutaneously on the backs of rats for 2 and 8 weeks. After sacrifice, implants and surrounding tissue were histologically processed. Light microscopic and histomorphometric evaluation of capsule thickness, capsule quality, and implant tissue interface was performed. In addition, we stained for alpha-smooth muscle actin (SMA), collagen, and ED-1 (a monocyte-macrophage marker). All implants were surrounded by a fibrous capsule. Capsules of the implants loaded with 1 or 2.5 microg of TGF-beta(3) showed significantly higher capsule quality. This meant that capsules were more mature compared with implants without TGF-beta(3). However, no significant differences were found in terms of thickness of the capsules or quality of the interface. Finally, apparently significant differences were also found in the expression of alpha-SMA, when comparing the various growth factor concentrations at both implantation points. In conclusion, the use of microgrooved PLA substrates with TGF-beta(3) did not lead to an overall improvement of periimplant tissue healing. PMID- 12625961 TI - Human endothelial progenitor cell-seeded hybrid graft: proliferative and antithrombogenic potentials in vitro and fabrication processing. AB - In this article, we show that human endothelial progenitor cells (EPCs) in circulating peripheral blood may be a novel cell source for a cell-incorporated engineered vascular graft. Cultures of human peripheral blood mononuclear cells collected by the density gradient technique developed highly proliferative EPC colonies, which expanded with culture time. The production rates of antiplatelet substances such as endothelial-type nitric oxide synthase and 6-keto prostaglandin-F(1)-alpha were approximately one-third and approximately one-half of those of mature endothelial cells (ECs), respectively. On the other hand, the tissue-type plasminogen activator production rate of EPCs was almost the same as that of ECs. EPCs were seeded and cultured on a small-diameter compliant graft (inner diameter, 1.5 mm) made of microporous segmented polyurethane film coated with a photo-reactive gelatin layer, and subsequently subjected to hydrodynamic shear stress by ex vivo circulation. EPCs fully covering the graft elongated and aligned themselves with the direction of the flow, resulting in the production of an integrated EPC-engineered graft. These results indicate that EPCs, which have high proliferative potential and high antithrombogenic potential, comparable to those of ECs, are a suitable cell source for cardiovascular tissue engineering. PMID- 12625962 TI - Efficient and stable retroviral transfection of ovine endothelial cells with green fluorescent protein for cardiovascular tissue engineering. AB - To determine whether cellular components of tissue-engineered cardiovascular structures are derived from cells harvested and seeded onto an acellular scaffold, or from cells originating from surrounding tissue (e.g., proximal and distal anastomosis), cellular retroviral transfection with green fluorescent protein (GFP) was used. Ovine endothelial cells (ECs) were transfected with a Moloney murine leukemia virus (Mo-MuLV)-based retroviral vector expressing GFP. Transfection was evaluated by fluorescence microscopy and fluorescence-activated cell sorting. The rate of transfection of the primary cells was 33.4% for ECs, 48 hours after transfection. Stable transfection could be observed for at least 25 subsequent passages. Retroviral transfection with GFP enables stable and reliable long-term labeling of ovine ECs. This approach might offer an attractive pathway to study tissue development, with emphasis on distinguishing between cellular components initially seeded onto a construct and those occurring as a result of cell ingrowth from surrounding tissue. PMID- 12625963 TI - Muscle cell-mediated gene delivery to the rotator cuff. AB - Rotator cuff lesions are one of the most common causes of upper extremity disability. Surgical therapy addresses mostly the extrinsic etiology, but not intrinsic factors such as aging, structural changes, low vascularity, and inflammatory processes. In this study, genetically engineered, highly purified muscle-derived cells (MDCs) were characterized and injected into the supraspinatus tendons of nude rats. The injected cells were monitored for 3 weeks. In vitro, the engineered, highly purified MDCs do not express vimentin; 98% of them are positive for the beta-galactosidase marker gene, and 99% hybridize with the specific pancentromeric mouse probe. beta-Galactosidase marker gene expression of the injected cells was detected up to 21 days. From day 7 after injection, the cell nuclei became spindle shaped, cells were integrated into the tendon collagen bundles, and the cells showed differentiation into vimentin-expressing fibroblastic cells. The results indicate that the rotator cuff tendon matrix and its original cellular components modulated the injected MDCs toward a fibroblastic phenotype. The compatibility and ability of MDCs to differentiate into other cell lineages, such as fibroblasts, might have high potential utility in tissue-engineering applications for tendon healing. This approach facilitates the application of muscle-derived progenitor cells and ex vivo gene therapy for the treatment of rotator cuff lesions. PMID- 12625964 TI - Novel approach to regeneration of periodontal tissues based on in situ tissue engineering: effects of controlled release of basic fibroblast growth factor from a sandwich membrane. AB - To regenerate periodontal tissues, a sandwich membrane composed of a collagen sponge scaffold and gelatin microspheres containing basic fibroblast growth factor (bFGF) in a controlled-release system was developed according to the new concept of "in situ tissue engineering." A three-walled alveolar bone defect (3 x 4 x 4 mm) was made bilaterally in edentulous regions created mesially to the canines in both the maxilla and mandible of nine beagle dogs. A sandwich membrane with or without bFGF (100 microg) was implanted in each defect (each group, n = 18). During weeks 1, 2, and 4, histologic evaluation and histometric analyses were performed on three dogs. Throughout the 4 weeks, vascularization and osteogenesis were active only in the bFGF-treated group (p < 0.01). New cementum was formed (2.4 +/- 0.9 mm) on the exposed root surface at 4 weeks, and functional recovery of the periodontal ligament was indicated in part by the perpendicular orientation of regenerated collagen fibers. In the control group, epithelial downgrowth and root resorption occurred and the defects were filled with connective tissue. Thus, our sandwich membrane induced successful regeneration of the periodontal tissues in a short period of time. PMID- 12625965 TI - Evaluation of transplanted tissue-engineered oral mucosa equivalents in severe combined immunodeficient mice. AB - The aim of this study was to determine the optimal stage of development at which transplant human ex vivo-produced oral mucosa equivalents (EVPOMEs) in vivo. EVPOMEs were generated in a serum-free culture system, without the use of an irradiated xenogeneic feeder layer, by seeding human oral keratinocytes onto a human cadaveric dermal equivalent, AlloDerm. EVPOMEs were cultured for 4 days submerged and then for 7 or 14 days at an air-liquid interface to initiate stratification before transplantation into SCID mice. AlloDerm, without epithelium, was used as a control. Mice were killed on days 3, 10, and 21 posttransplantation. Epithelium of the transplanted EVPOMEs was evaluated with the differentiation marker keratin 10/13. Dermal microvessel ingrowth was determined by immunohistochemistry with a mouse vascular marker, lectin binding from Triticum vulgaris. The presence and stratification of the epithelium were correlated with revascularization of the underlying dermis. The microvessel density of AlloDerm without epithelium was less than that of EVPOMEs with an epithelial layer. Microvessel density of the dermis varied directly with the degree of epithelial stratification of the EVPOMEs. The EVPOMEs cultured at an air-liquid interface for 7 days had the optimal balance of neoangiogenesis and epithelial differentiation necessary for in vivo grafting. PMID- 12625966 TI - Growth of porcine enamel-, dentin-, and cementum-derived cells in collagen glycosaminoglycan matrices in vitro: expression of alpha-smooth muscle actin and contraction. AB - The objective of the study was to investigate the behavior of porcine enamel, dentin, and cementum cells, isolated from tissue digests and growing out from explants, in monolayer culture and in a collagen-glycosaminoglycan (GAG) matrix for tissue engineering. A notable finding of the study was the expression of a contractile muscle actin isoform, alpha-smooth muscle actin (SMA), by each cell type and their contraction of the collagen-GAG scaffold. Of importance was the immunohistochemical demonstration that the majority of ameloblasts and odontoblasts in vivo contained SMA. Western blot analysis demonstrated the presence of SMA in all of the cell types. A large amount of SMA was found in the odontoblasts after the first passage. SMA expression in the enamel- and cementum derived cells appeared to increase with time in culture and with passage number. The implications of this finding for tissue engineering and the possible roles for SMA in dental tissue cells are discussed. PMID- 12625967 TI - Antimicrobial susceptibility of Gram-negative uropathogens isolated from obstetric patients. AB - OBJECTIVE: To evaluate the antimicrobial susceptibility of Gram-negative uropathogens isolated from pregnant women. METHODS: We performed a snapshot cohort study of women receiving care in the University of Florida prenatal clinics during March 2000. Subjects with asymptomatic bacteriuria or cystitis were identified and the antimicrobial susceptibility of each pathogen was recorded. Data were analyzed using chi-square, Fisher's exact test and ninety five percent confidence intervals, as appropriate. RESULTS: Ninety-five positive cultures were identified. Isolates were more often susceptible to trimethoprim sulfamethoxazole (TMP-SMX) (87%) and nitrofurantoin (89%) than to ampicillin (72%) (p < 0.03). Escherichia coli accounted for 71 (75%) cases and was more often susceptible to nitrofurantoin (100%) than to TMP-SMX (87%) (p < 0.01). Proteus isolates were all susceptible to TMP-SMX and resistant to nitrofurantoin (p < 0.01). CONCLUSIONS: Both TMP-SMX and nitrofurantoin are superior to ampicillin for empiric treatment of lower urinary tract infection in pregnant women. Nitrofurantoin is superior to TMP-SMX for treatment of infections caused by E. coli. For suspected or confirmed cases caused by Proteus organisms, TMP-SMX is the preferred agent. PMID- 12625968 TI - Risk of perinatal transmission of hepatitis B virus in Jordan. AB - OBJECTIVES: To determine the risk of perinatal transmission of hepatitis B virus (HBV) in Jordan. METHODS: Plasma samples from 1000 pregnant Jordanian women were screened by ELISA for HBV markers (HBsAg, HBeAg, anti-HBe, anti-HBc and anti HBs). RESULTS: HBsAg and HBeAg were detected in 4.3% and 0.1% of the pregnant women, respectively. The overall prevalence of antibodies was 6%, 11.1% and 7.5% for anti-HBe, anti-HBc and anti-HBs, respectively. Women were assigned to four groups according to the serological patterns of HBV markers: susceptible (85.9%), with acute infection (2.9%), with chronic infection (1.4%) and previously infected (9.8%). Most women were at the third trimester of pregnancy, therefore women with acute and chronic hepatitis at this gestational age were at risk of transmitting HBV infection to their newborns. Women who belonged to the low socio economic class were at higher risk of HBV infection. CONCLUSIONS: Based on the results, we recommend screening women for HBV during pregnancy in order to identify HBV carriers. All newborns born to carriers should be vaccinated immediately after birth, both passively and actively. Also vaccination of HBV seronegative pregnant women is recommended. PMID- 12625970 TI - Site-specific prevalence and cell densities of selected microbes in the lower reproductive tract of menstruating tampon users. AB - OBJECTIVE: To assess differences in prevalence and cell densities of enterococci, Gram negative enterics (GNEs), yeast and Staphylococcus aureus among four genital sites and to examine whether the presence of organisms at one site affected the presence of organisms at other sites. METHODS: Swab samples from the perineum, below and above the hymen, and the posterior fornix obtained from 52 tampon users on menstrual cycle day 3 were analyzed for site-specific prevalence and cell densities of microorganisms. RESULTS: Enterococci and GNEs were the most prevalent study organisms at all sites and decreased in prevalence from the perineum to the posterior fornix. Cell densities similarly decreased from below the hymen to the posterior fornix. Yeast were detected at the hymen only; S. aureus frequency was similarly low at all sites. Yeast and S. aureus site specific cell densities were similar. The above- and below-hymen sites were similar in prevalence and cell density of organisms. An above-chance association existed between the presence of any study organism below the hymen and above the hymen and was strongest for GNEs. CONCLUSIONS: The pattern of genital colonization with enterococci and GNEs reflects their likely gastrointestinal source. The absence of significant differences in the prevalence and cell densities of study microflora above and below the hymen combined with an above chance association of the presence of microorganisms at these regions suggests that the regions above and below the hymen are not different with respect to the presence of the organisms evaluated in this study. PMID- 12625971 TI - Vulvar myiasis during pregnancy. AB - Myiasis is a parasitic infestation caused by the larvae of several fly species. Diagnosis and treatment are simple. The location of this infestation at the vulvar area is, however, an extremely rare occurrence. The authors present two cases of vulvar myiasis affecting pregnant women. The first case is a 19-year-old pregnant girl with vulvar myiasis and concomitant syphilis, vaginal trichomoniasis and genital candidiasis. The patient was also HIV-positive. The second case is a 17-year-old pregnant girl with vulvar myiasis associated with extensive vulvar condyloma acuminatum lesions. PMID- 12625969 TI - Does pre- and postoperative metronidazole treatment lower vaginal cuff infection rate after abdominal hysterectomy among women with bacterial vaginosis? AB - OBJECTIVE: Bacterial vaginosis (BV) is a known risk factor for postoperative infection following abdominal hysterectomy. Vaginal bacterial flora scored as intermediate has been shown to have the same risk of postoperative infection as BV. METHODS: Women undergoing total abdominal hysterectomy for benign diseases were open-randomized according to Zelen to either treatment with metronidazole rectally for at least 4 days or no treatment. At the preoperative gynecological examination a vaginal smear was collected and Gram stained. Women with BV or intermediate flora were merged to one group called abnormal vaginal flora. RESULTS: In total 213 women were randomized to treatment or no treatment. After exclusion of 71 women, 142 women were eligible for analysis. Among the 59 women diagnosed with abnormal vaginal flora there were no vaginal cuff infections in the treated arm, compared with 27% in the 'no treatment' arm (p < 0.01). Treatment also reduced the vaginal cuff infection rate from 9.5 to 2% among the 83 women with lactobacilli flora. However, this difference was not statistically significant. Treatment had no effect on the rate of wound infections. Intention to-treat analysis showed a significant reduction in vaginal cuff infections among women randomized to treatment. CONCLUSION: Pre- and postoperative treatment for at least 4 days with metronidazole rectally reduces significantly vaginal cuff infection among women with abnormal vaginal flora. PMID- 12625972 TI - Varicella vaccination during early pregnancy: a cause of in utero miliary fetal tissue calcifications and hydrops? AB - BACKGROUND: It is the purpose of this article to describe a suspected association of inadvertent vaccination with varicella vaccine during early pregnancy with the subsequent development of in utero miliary fetal tissue calcifications and fetal hydrops detected by sonogram at 15 weeks of gestation. CASE: This is a case presentation of a pregnant patient who received varicella vaccination during the same menstrual cycle that she became pregnant, and is supplemented by a literary review. The fetus developed miliary fetal tissue calcifications and fetal hydrops detected by a targeted sonogram at 15 weeks gestation. CONCLUSION: Varicella vaccination during early pregnancy may be a cause of miliary fetal tissue calcifications and fetal hydrops. PMID- 12625973 TI - Haemophilus influenzae septic abortion. AB - BACKGROUND: Haemophilus influenzae septic abortion is typically caused by nontypeable strains of the organism. Furthermore, nontypeable species with a special affinity for the genital tract are the most frequent isolates encountered, and an ascending vaginal or cervical infection is often the suspected route of transmission. CASE: A 39-year-old woman at 8 weeks gestation who underwent dilation, evacuation, and curettage for embryonic demise had clinical evidence for sepsis and isolation of a nontypeable, ampicillin resistant H. influenzae from blood cultures. Although an ascending vaginal infection was suspected, the route of transmission was not determined. CONCLUSION: Nontypeable strains of. H. influenzae have demonstrated increased beta-lactamase activity, and ampicillin, formerly the treatment of choice, should be used only if isolate susceptibility is known. PMID- 12625976 TI - Laboratory medicine practice guidelines. Laboratory support for the diagnosis and monitoring of thyroid disease. PMID- 12625990 TI - Methylphenidate improves Stroop naming speed, but not response interference, in children with attention deficit hyperactivity disorder. AB - OBJECTIVE: The goal of this study was to investigate the effect of methylphenidate (MPH) on response interference, as measured by the Stoop Color and Word Test, in children with attention deficit hyperactivity disorder (ADHD). Response interference is a core component of response inhibition that has been shown to be impaired in children with ADHD. METHODS: A clinic-referred sample of school-aged children with a confirmed Diagnostic and Statistical Manual of Mental Disorders (4th ed.) diagnosis of ADHD and good reading skills (n = 31) completed the Stroop Color and Word Test in an acute, randomized, placebo-controlled, crossover trial with three single fixed doses of MPH. RESULTS: MPH did not improve response interference on the Stroop Color and Word Test but did significantly improve color naming and word naming abilities. CONCLUSION: Response interference, as measured by the Stroop Color and Word Test, is not improved by MPH in children with ADHD. In addition, findings demonstrate strongly positive MPH effects on the highly effortful process of color naming, which has previously been demonstrated as impaired in children with ADHD. MPH was also shown to have a positive but smaller effect on word naming speed. PMID- 12625991 TI - Prevalence and patterns of use of psychoactive medicines in individuals with autism in the Autism Society of North Carolina. AB - The aim of this study was to assess the prevalence and patterns of psychoactive and over-the-counter medicines in a large cohort of individuals with autism. We conducted a mail survey of 3,228 families that are members of the Autism Society of North Carolina. This is one of the largest chapters of the Autism Society of America. The survey form addressed current medicines used, side effects, demographic characteristics, and medical conditions. Some 1,538 member families within the society (48%) responded to the survey. In all, 703 (45.7%) individuals with autism were taking psychotropic drugs, 191 (12.4%) antiepileptic drugs (AEDs), and 86 (5.7%) supplements for autism. The total number taking psychotropic, antiepileptic, or vitamin treatments was 816 (53.1%). Antidepressants (taken by 21.7% of the sample), antipsychotics (16.8%), and stimulants (13.9%) were the most commonly prescribed agents. Univariate and multivariate analyses were conducted to examine factors associated with treatment. Greater age, more severe autism and mental retardation, and more restricted housing were often associated with greater use of psychoactive agents. These findings suggest that individuals with autism are a frequently medicated group, although the empirical research support for most agents being used is still very limited. PMID- 12625992 TI - Adjunctive topiramate treatment for pediatric bipolar disorder: a retrospective chart review. AB - OBJECTIVE: The objective of this study was to evaluate the effectiveness, safety, and tolerability of the anticonvulsant agent, topiramate, as adjunctive treatment for children and adolescents with bipolar disorders. METHODS: The outpatient medical charts of children and adolescents with a Diagnostic and Statistical Manual of Mental Disorders (4th ed.) diagnosis of bipolar disorder, type I or II, and who were treated with topiramate were retrospectively reviewed by two child and adolescent psychiatrists using the Clinical Global Impression (CGI) scale and the Clinical Global Assessment Scale (CGAS). Separate CGI ratings were made for mania and overall bipolar illness. RESULTS: Twenty-six patients (mean age 14 +/- 3.5 years) with bipolar disorder, type I (n = 23) or II (n = 3), who had been treated (mean duration 4.1 +/- 6.1 months) with topiramate (mean dose 104 +/- 77 mg/day) were identified. Response rate (defined by a CGI-Improvement score of < or = 2 at endpoint) was 73% for mania and 62% for overall illness. CGAS scores significantly improved from baseline to endpoint. No serious adverse events were reported. CONCLUSIONS: Although controlled trials are necessary, this retrospective study suggests that topiramate is effective and well tolerated as an adjunctive treatment for children and adolescents with bipolar disorder. PMID- 12625993 TI - Glutamatergic changes with treatment in attention deficit hyperactivity disorder: a preliminary case series. AB - Magnetic resonance spectroscopy, a noninvasive neuroimaging method, is a technique with the potential to measure in vivo neurochemical changes to different medication treatments. Symptoms of attention deficit hyperactivity disorder (ADHD) improved in two children treated with methylphenidate and two children treated with atomoxetine, for whom pre- and posttreatment proton magnetic resonance spectroscopy examinations were obtained to assess the relation between the neurochemical profiles in the striatum and prefrontal cortex among symptom severity and response to treatment. In the striatum, a striking decrease in the glutamate/creatine ratio (mean change 56.1%) was observed between 14 and 18 weeks of therapy in all four children with ADHD. In the prefrontal cortex, however, changes in the glutamate/creatine ratio were noted only in subjects receiving atomoxetine, not in those receiving methylphenidate. These data suggest that in vivo magnetic resonance spectroscopy measurement has the potential to assess response to psychopharmacological treatment in children with ADHD. PMID- 12625994 TI - A case series of eight aggressive young children treated with risperidone. AB - The aim of this study was to examine the use of risperidone in routine clinical care for very aggressive young children. This is a retrospective medical chart review of patients age less than 6 years 11 months who were treated with risperidone for 1 to 10 months during the 1-year study period. Treatment response, side effects, and Clinical Global Impression (CGI) scores were identified. One hundred and five such young children were identified; 8 had been treated with risperidone (6 boys, 2 girls: mean age 4.9 +/- 0.8 years). Risperidone was used in combination with other psychotropic medications in 7 of the 8 children. The mean daily dose of risperidone was 1.25 +/- 0.27 mg. Seventy five percent of the children were on concomitant lithium, valproate, or carbamazepine; 63% were on stimulants or alpha adrenergics. This was a highly comorbid group, with 7 children presenting with attention deficit hyperactivity disorder and 5 children with bipolar disorder not otherwise specified. The average baseline CGI severity was 5.5 (SD = 0.5), and at last visit it was 3.5 (SD = 0.5), p < 0.0001. Mean CGI improvement score was 1.9 (SD = 0.6). Adverse effects included significant weight gain (mean 5.5 +/- 4.9 kg, p < 0.05) in 6 patients. One child had hyperprolactinemia. Given the potential development of atherosclerosis in obesity and endocrine response in hyperprolactinemia, risperidone should be reserved for those children with severe aggressive behavior who failed multiple trials with other agents. Further controlled trials are needed. PMID- 12625995 TI - Clozapine-induced akathisia in children with schizophrenia. AB - Akathisia is a relatively rare side effect with the newer atypical antipsychotic agents, particularly clozapine, and is easily misdiagnosed in children. As children are often unable to describe their symptoms verbally, their akathisia can be misdiagnosed as worsening of their psychosis, prompting an unnecessary increase in their neuroleptic dose. Two cases of childhood-onset schizophrenia associated with clozapine-induced akathisia responsive to beta-blocker treatment are described. Akathisia should be considered in all cases of apparent nonresponse to atypical antipsychotics. PMID- 12625996 TI - Olanzapine overdose: a pediatric case report. AB - This paper reports a case of an olanzapine overdose in a 12-year-old boy. An opioid-like presentation was noted. Despite a high serum level of olanzapine, the patient made a complete recovery and showed no sequelae at follow-up. PMID- 12625997 TI - Pediatric quetiapine overdose: a case report and literature review. AB - Quetiapine is a medication approved for the treatment of psychotic disorders in adults. At this time it is not approved for the treatment of children or adolescents. It is an atypical antipsychotic agent that is efficacious in treating both the positive and negative symptoms of schizophrenia. There is currently little information available concerning the safety of quetiapine in overdose, and there are no previous case reports of quetiapine overdose in the pediatric population. We present the case of a 15-year-old girl who ingested 1250 mg of quetiapine (21.6 mg/kg) in a suicide attempt. She developed multiple symptoms including tachycardia, agitation, hypotension, and unconsciousness. We compare her symptoms to previous adult cases of quetiapine overdose and review overdose treatment recommendations. We also examine clinical situations that may lead to a more severe clinical course. PMID- 12625998 TI - Use of nortriptyline in adolescent-onset bipolar disorder. PMID- 12625999 TI - Lamotrigine and borderline personality disorder. PMID- 12626000 TI - Sexual disinhibition induced by risperidone in a child. PMID- 12626001 TI - The initiation of bursts in thalamic neurons and the cortical control of thalamic sensitivity. AB - Thalamic neurons generate high-frequency bursts of action potentials when a low threshold (T-type) calcium current, located in soma and dendrites, becomes activated. Computational models were used to investigate the bursting properties of thalamic relay and reticular neurons. These two types of thalamic cells differ fundamentally in their ability to generate bursts following either excitatory or inhibitory events. Bursts generated with excitatory inputs in relay cells required a high degree of convergence from excitatory inputs, whereas moderate excitation drove burst discharges in reticular neurons from hyperpolarized levels. The opposite holds for inhibitory rebound bursts, which are more difficult to evoke in reticular neurons than in relay cells. The differences between the reticular neurons and thalamocortical neurons were due to different kinetics of the T-current, different electrotonic properties and different distribution patterns of the T-current in the two cell types. These properties enable the cortex to control the sensitivity of the thalamus to inputs and are also important for understanding states such as absence seizures. PMID- 12626002 TI - Thalamic circuitry and thalamocortical synchrony. AB - The corticothalamic system has an important role in synchronizing the activities of thalamic and cortical neurons. Numerically, its synapses dominate the inputs to relay cells and to the gamma-amino butyric acid (GABA)ergic cells of the reticular nucleus (RTN). The capacity of relay neurons to operate in different voltage-dependent functional modes determines that the inputs from the cortex have the capacity directly to excite the relay cells, or indirectly to inhibit them via the RTN, serving to synchronize high- or low-frequency oscillatory activity respectively in the thalamocorticothalamic network. Differences in the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) subunit composition of receptors at synapses formed by branches of the same corticothalamic axon in the RTN and dorsal thalamus are an important element in the capacity of the cortex to synchronize low-frequency oscillations in the network. Interactions of focused corticothalamic axons arising from layer VI cortical cells and diffuse corticothalamic axons arising from layer V cortical cells, with the specifically projecting core relay cells and diffusely projecting matrix cells of the dorsal thalamus, form a substrate for synchronization of widespread populations of cortical and thalamic cells during high-frequency oscillations that underlie discrete conscious events. PMID- 12626005 TI - Is there a thalamic component to experience-dependent cortical plasticity? AB - Sensory deprivation and injury to the peripheral nervous system both induce plasticity in the somatosensory system of adult animals, but in different places. While injury induces plasticity at several locations within the ascending somatosensory pathways, sensory deprivation appears only to affect the somatosensory cortex. Experiments have been performed to detect experience dependent plasticity in thalamic receptive fields, thalamic domain sizes and convergence of thalamic receptive fields onto cortical cells. So far, plasticity has not been detected with sensory deprivation paradigms that cause substantial cortical plasticity. Part of the reason for the lack of thalamic plasticity may lie in the synaptic properties of afferent systems to the thalamus. A second factor may lie in the differences in the organization of cortical and thalamic circuits. Many deprivation paradigms induce plasticity by decreasing phasic lateral inhibition. Since lateral inhibition appears to be far weaker in the thalamus than the cortex, sensory deprivation may not cause large enough imbalances in thalamic activity to induce plasticity in the thalamus. PMID- 12626003 TI - Novel neuronal and astrocytic mechanisms in thalamocortical loop dynamics. AB - In this review, we summarize three sets of findings that have recently been observed in thalamic astrocytes and neurons, and discuss their significance for thalamocortical loop dynamics. (i) A physiologically relevant 'window' component of the low-voltage-activated, T-type Ca(2+) current (I(Twindow)) plays an essential part in the slow (less than 1 Hz) sleep oscillation in adult thalamocortical (TC) neurons, indicating that the expression of this fundamental sleep rhythm in these neurons is not a simple reflection of cortical network activity. It is also likely that I(Twindow) underlies one of the cellular mechanisms enabling TC neurons to produce burst firing in response to novel sensory stimuli. (ii) Both electrophysiological and dye-injection experiments support the existence of gap junction-mediated coupling among young and adult TC neurons. This finding indicates that electrical coupling-mediated synchronization might be implicated in the high and low frequency oscillatory activities expressed by this type of thalamic neuron. (iii) Spontaneous intracellular Ca(2+) ([Ca(2+)](i)) waves propagating among thalamic astrocytes are able to elicit large and long-lasting N-methyl-D-aspartate-mediated currents in TC neurons. The peculiar developmental profile within the first two postnatal weeks of these astrocytic [Ca(2+)](i) transients and the selective activation of these glutamate receptors point to a role for this astrocyte-to-neuron signalling mechanism in the topographic wiring of the thalamocortical loop. As some of these novel cellular and intracellular properties are not restricted to thalamic astrocytes and neurons, their significance may well apply to (patho)physiological functions of glial and neuronal elements in other brain areas. PMID- 12626004 TI - The role of the thalamus in the flow of information to the cortex. AB - The lateral geniculate nucleus is the best understood thalamic relay and serves as a model for all thalamic relays. Only 5-10% of the input to geniculate relay cells derives from the retina, which is the driving input. The rest is modulatory and derives from local inhibitory inputs, descending inputs from layer 6 of the visual cortex, and ascending inputs from the brainstem. These modulatory inputs control many features of retinogeniculate transmission. One such feature is the response mode, burst or tonic, of relay cells, which relates to the attentional demands at the moment. This response mode depends on membrane potential, which is controlled effectively by the modulator inputs. The lateral geniculate nucleus is a first-order relay, because it relays subcortical (i.e. retinal) information to the cortex for the first time. By contrast, the other main thalamic relay of visual information, the pulvinar region, is largely a higher-order relay, since much of it relays information from layer 5 of one cortical area to another. All thalamic relays receive a layer-6 modulatory input from cortex, but higher-order relays in addition receive a layer-5 driver input. Corticocortical processing may involve these corticothalamocortical 're-entry' routes to a far greater extent than previously appreciated. If so, the thalamus sits at an indispensable position for the modulation of messages involved in corticocortical processing. PMID- 12626006 TI - Thalamocortical control of feed-forward inhibition in awake somatosensory 'barrel' cortex. AB - Intracortical inhibition plays a role in shaping sensory cortical receptive fields and is mediated by both feed-forward and feedback mechanisms. Feed-forward inhibition is the faster of the two processes, being generated by inhibitory interneurons driven by monosynaptic thalamocortical (TC) input. In principle, feed-forward inhibition can prevent targeted cortical neurons from ever reaching threshold when TC input is weak. To do so, however, inhibitory interneurons must respond to TC input at low thresholds and generate spikes very quickly. A powerful feed-forward inhibition would sharpen the tuning characteristics of targeted cortical neurons, and interneurons with sensitive and broadly tuned receptive fields could mediate this process. Suspected inhibitory interneurons (SINs) with precisely these properties are found in layer 4 of the somatosensory (S1) 'barrel' cortex of rodents and rabbits. These interneurons lack the directional selectivity seen in most cortical spiny neurons and in ventrobasal TC afferents, but are much more sensitive than cortical spiny neurons to low amplitude whisker displacements. This paper is concerned with the activation of S1 SINs by TC impulses, and with the consequences of this activation. Multiple TC neurons and multiple S1 SINs were simultaneously studied in awake rabbits, and cross-correlation methods were used to examine functional connectivity. The results demonstrate a potent, temporally precise, dynamic and highly convergent/divergent functional input from ventrobasal TC neurons to SINs of the topographically aligned S1 barrel. Whereas the extensive pooling of convergent TC inputs onto SINs generates sensitive and broadly tuned inhibitory receptive fields, the potent TC divergence onto many SINs generates sharply synchronous activity among these elements. This TC feed-forward inhibitory network is well suited to provide a fast, potent, sensitive and broadly tuned inhibition of targeted spiny neurons that will suppress spike generation following all but the most optimal feed-forward excitatory inputs. PMID- 12626007 TI - Spike timing and visual processing in the retinogeniculocortical pathway. AB - Although the visual response properties of neurons along the retinogeniculocortical pathway have been studied for decades, relatively few studies have examined how individual neurons along the pathway communicate with each other. Recent studies in the cat (Felis domestica) now show that the strength of these connections is very dynamic and spike timing plays an important part in determining whether action potentials will be transferred from pre- to postsynaptic cells. This review explores recent progress in our understanding of what role spike timing has in establishing different patterns of geniculate activity and how these patterns ultimately drive the cortex. PMID- 12626009 TI - Dynamic shifting in thalamocortical processing during different behavioural states. AB - Recent experiments in our laboratory have indicated that as rats shift the behavioural strategy employed to explore their surrounding environment, there is a parallel change in the physiological properties of the neuronal ensembles that define the main thalamocortical loop of the trigeminal somatosensory system. Based on experimental evidence from several laboratories, we propose that this concurrent shift in behavioural strategy and thalamocortical physiological properties provides rats with an efficient way to optimize either the detection or analysis of complex tactile stimuli. PMID- 12626008 TI - Corticothalamic interactions in the transfer of visual information. AB - Thalamic function does not stand apart, as a discrete processing step, from the cortical circuitry. The thalamus receives extensive feedback from the cortex and this influences the firing pattern, synchronization and sensory response mode of relay cells. A crucial question concerns the extent to which the feedback simply controls the state and transmission mode of relay cells and the extent to which the feedback participates in the specific processing of sensory information. Using examples from experiments examining the influence of feedback from the visual cortex to the lateral geniculate nucleus (LGN), we argue that thalamic mechanisms are selectively focused by visually driven feedback to optimize the thalamic contribution to segmentation and global integration. This involves effects on both the temporal and spatial parameters characterizing the responses of LGN cells and includes, for example, motion-driven feedback effects from MT (middle temporal visual area) relayed via layer 6 of V1 (primary visual cortex). PMID- 12626010 TI - Glutamate receptor functions in sensory relay in the thalamus. AB - It is known that glutamate is a major excitatory transmitter of sensory and cortical afferents to the thalamus. These actions are mediated via several distinct receptors with postsynaptic excitatory effects predominantly mediated by ionotropic receptors of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) and N-methyl-D-aspartate varieties (NMDA). However, there are also other kinds of glutamate receptor present in the thalamus, notably the metabotropic and kainate types, and these may have more complex or subtle roles in sensory transmission. This paper describes recent electrophysiological experiments done in vitro and in vivo which aim to determine how the metabotropic and kainate receptor types can influence transmission through the sensory thalamic relay. A particular focus will be how such mechanisms might operate under physiological conditions. PMID- 12626011 TI - A new interpretation of thalamocortical circuitry. AB - Almost all the information that is needed to specify thalamocortical and neocortical wiring derives from patterned electrical activity induced by the environment. Wiring accuracy must be limited by the anatomical specificity of the cascade of events triggered by neural activity and culminating in synaptogenesis. We present a simple model of learning in the presence of plasticity errors. One way to achieve learning specificity is to build better synapses. We discuss an alternative, circuit-based, approach that only allows plasticity at connections that support highly selective correlations. This circuit resembles some of the more puzzling aspects of thalamocorticothalamic circuitry. PMID- 12626012 TI - Target and temporal pattern selection at neocortical synapses. AB - We attempt to summarize the properties of cortical synaptic connections and the precision with which they select their targets in the context of information processing in cortical circuits. High-frequency presynaptic bursts result in rapidly depressing responses at most inputs onto spiny cells and onto some interneurons. These 'phasic' connections detect novelty and changes in the firing rate, but report frequency of maintained activity poorly. By contrast, facilitating inputs to interneurons that target dendrites produce little or no response at low frequencies, but a facilitating-augmenting response to maintained firing. The neurons activated, the cells they in turn target and the properties of those synapses determine which parts of the circuit are recruited and in what temporal pattern. Inhibitory interneurons provide both temporal and spatial tuning. The 'forward' flow from layer-4 excitatory neurons to layer 3 and from 3 to 5 activates predominantly pyramids. 'Back' projections, from 3 to 4 and 5 to 3, do not activate excitatory cells, but target interneurons. Despite, therefore, an increasing complexity in the information integrated as it is processed through these layers, there is little 'contamination' by 'back' projections. That layer 6 acts both as a primary input layer feeding excitation 'forward' to excitatory cells in other layers and as a higher-order layer with more integrated response properties feeding inhibition to layer 4 is discussed. PMID- 12626013 TI - Excitatory inputs to spiny cells in layers 4 and 6 of cat striate cortex. AB - The principal target of lateral geniculate nucleus in the cat visual cortex is the stellate neurons of layer 4. In previously reported work with intracellular recording and extracellular stimulation in slices of visual cortex, three general classes of fast excitatory synaptic potentials (EPSPs) in layer 4a spiny stellate neurons were identified. One of these classes, characterized by large and relatively invariant amplitudes (mean 1.7 mV, average coefficient of variation (CV) 0.083) were attributed to the action of geniculate axons because, unlike the other two classes, they could not be matched by intracortical inputs, using paired recording. We have examined in detail the properties of this synaptic input in twelve examples, selecting for study those EPSPs where there was secure extracellular stimulation of the single fibre input to a pair of stimuli 50 ms apart. In our analysis, we conclude that the depression that these inputs show to the second stimulus is entirely postsynaptic, since the evidence strongly suggests that the probability of transmitter release at the synaptic site(s) remains 1.0 for both stimuli. We argue that the most plausible explanation for this postsynaptic depression is a reduction in the average probability of opening the synaptic channels. Using a simple biochemical analysis (c.f. Sigworth plot), it is then possible to calculate the number of synaptic channels and their probability of opening, for each of the 12 connections. The EPSPs had a mean amplitude of 1.91 mV (+/- 1.3 mV SD) and a mean CV of 0.067 (+/- 0.022). The calculated number of channels ranged from 20 to 158 (59.4 +/- 48.7) and their probability of opening to the first EPSP had an average of 0.83 (+/- 0.09), with an average depression of the probability to 0.60 for the second EPSP. Geniculate afferents also terminate in layer 6. Intracellular recordings were also made in the upper part of this layer and a total of 51 EPSPs were recorded from pyramidal cells of three principal types. Amongst this dataset we sought EPSPs with similar properties to those characterized in layer 4a. Three examples were found, which is a much lower percentage (6%) than the incidence of putative geniculate EPSPs found in layer 4a (42%). PMID- 12626014 TI - The thalamus as a monitor of motor outputs. AB - Many of the ascending pathways to the thalamus have branches involved in movement control. In addition, the recently defined, rich innervation of 'higher' thalamic nuclei (such as the pulvinar) from pyramidal cells in layer five of the neocortex also comes from branches of long descending axons that supply motor structures. For many higher thalamic nuclei the clue to understanding the messages that are relayed to the cortex will depend on knowing the nature of these layer five motor outputs and on defining how messages from groups of functionally distinct output types are combined as inputs to higher cortical areas. Current evidence indicates that many and possibly all thalamic relays to the neocortex are about instructions that cortical and subcortical neurons are contributing to movement control. The perceptual functions of the cortex can thus be seen to represent abstractions from ongoing motor instructions. PMID- 12626015 TI - The influence of the corticothalamic projection on responses in thalamus and cortex. AB - We review results on the in vivo properties of neurons in the dorsal lateral geniculate nucleus (dLGN) that receives its afferent input from the retina and projects to the visual cortex. In addition, the dLGN receives input from the brain stem and from a rather strong corticothalamic back-projection, which originates in layer 6 of the visual cortex. We compare the behaviour of dLGN cells during spontaneous changes of the frequency contents of the electroencephalograph (EEG) (which are mainly related to a changing brain stem influence), with those that are obtained when experimentally silencing the corticothalamic feedback. The spatial and temporal response properties of dLGN cells are compared during these two conditions, and we report that the neurons behave similarly during a synchronized EEG state and during inactive corticothalamic feedback. In both situations, dLGN cells are rather phasic and their remaining tonic activity is temporally dispersed, indicating a hyperpolarizing effect. By means of a novel method, we were able to chronically eliminate a large proportion of the corticothalamic projection neurons from the otherwise intact cortex. In this condition, we found that cortical cells also lose their EEG specific response differences but, in this instance, probably due to a facilitatory (depolarizing) plasticity reaction of the remaining network. PMID- 12626016 TI - Recordings, behaviour and models related to corticothalamic feedback. AB - In this paper, we review recent work on aspects of corticothalamic interactions in the auditory and in the visual systems. There are gross similarities in the arrangements of these systems, but considerable contrasts in the processing computations and in the effects of corticothalamic feedback. PMID- 12626017 TI - Analysing neuronal correlates of the comparison of two sequentially presented sensory stimuli. AB - In a typical sequential sensory discrimination task, subjects are required to make a decision based on comparing a sensory stimulus against the memory trace left by a previous stimulus. What is the neuronal substrate for such comparisons and the resulting decisions? This question was studied by recording neuronal responses in a variety of cortical areas of awake monkeys (Macaca mulatta), trained to carry out a vibrotactile sequential discrimination task. We describe methods to analyse responses obtained during the comparison and decision phases of the task, and describe the resulting findings from recordings in secondary somatosensory cortical area (S2). A subset of neurons in S2 become highly correlated with the monkey's decision in the task. PMID- 12626018 TI - Rate and timing in cortical synaptic plasticity. AB - Debate has raged over the past few years as to whether cortical neurons transmit information primarily in their average firing rates or in the precise timing of their spikes. Here, we address the related question of which features of spike trains control plasticity at cortical synapses. Using paired recording in slices we have developed a quantitative and predictive description of the joint dependence of cortical plasticity on the rate and relative timing of pre- and postsynaptic firing. The results hold important implications for which parts of the neural code are most readily stored for later retrieval. PMID- 12626020 TI - Feed-forward synchronization: propagation of temporal patterns along the retinothalamocortical pathway. AB - Visual responses in the cortex and lateral geniculate nucleus (LGN) are often associated with synchronous oscillatory patterning. In this short review, we examine the possible relationships between subcortical and cortical synchronization mechanisms. Our results obtained from simultaneous multi-unit recordings show strong synchronization of oscillatory responses between retina, LGN and cortex, indicating that cortical neurons can be synchronized by oscillatory activity relayed through the LGN. This feed-forward synchronization mechanism operating in the 60 to 120 Hz frequency range was observed mostly for static stimuli. In response to moving stimuli, by contrast, cortical synchronization was independent of oscillatory inputs from the LGN, with oscillation frequency in the range of 30 to 60 Hz. The functional implications of synchronization of activity from parallel channels are discussed, in particular its significance for signal transmission and cortical integration processes. PMID- 12626019 TI - Adaptation in the corticothalamic loop: computational prospects of tuning the senses. AB - The present article discusses computational hypotheses on corticothalamic feedback and modulation of cortical response properties. We have recently proposed that the two phenomena are related, hypothesizing that neuronal velocity preference in the visual cortex is altered by feedback to the lateral geniculate nucleus. We now contrast the common view that response adaptation to stimuli subserves a function of redundancy reduction with the idea that it may enhance cortical representation of objects. Our arguments lead to the concept that the corticothalamic loop is involved in reducing sensory input to behaviourally relevant aspects, a pre-attentive gating. PMID- 12626021 TI - Pre-attentive segmentation and correspondence in stereo. AB - Traditional stereo grouping models have focused on the problem of stereo correspondence between monocular inputs. Recent physiological data revealed that the disparity selective V2 cells increase their responses when (random-dot stereograms) stimuli within their receptive fields are at or near the boundary of a depth surface. Such highlights to depth (non-luminance) edges are seemingly not computationally required for the correspondence problem. Computationally, these highlights make the boundaries of a depth surface more salient, serving pre attentive segmentation (between depth planes) and attracting visual attention. In special cases, they enable the psychophysically observed perceptual pop-out of a target from a background of visually identical distractors at a different depth. To achieve the highlights, mutual inhibition between disparity selective cells that are tuned to the same or similar depths is required. However, such mutual inhibition would impede the computation for the correspondence problem, which requires mutual excitation between the same cells. In this work, I introduce a computational model that, I believe, is the first to address both stereo correspondence and pre-attentive stereo segmentation. The computational mechanisms in the model are based on intracortical interactions in V2. I will demonstrate that the model captures the following physiological and psychophysical phenomena: (i) depth-edge highlighting; (ii) disparity capture; (iii) pop-out; and (iv) transparency. PMID- 12626023 TI - Hospital downsizing and workforce reduction strategies: some inner workings. AB - Downsizing, manpower reductions, re-engineering, and resizing are used extensively in the United States to reduce cost and to evaluate the effectiveness and efficiency of various functions and processes. Published studies report that these managerial strategies result in a minimal impact on access to services, quality of care, and the ability to reduce costs. But, these approaches certainly alienate employees. These findings are usually explained by the significant difficulties experienced in eliminating nursing and other similar direct patient care-oriented positions and in terminating white-collar employees. Possibly an equally plausible reason why hospitals and physician practices react so poorly to these management strategies is their cost structure-high fixed (85%) and low variable (15%)-and that simply generating greater volume does not necessarily achieve economies of scale. More workable alternatives for health executives to effectuate cost reductions consist of simplifying prepayment, decreasing the overall availability and centralizing tertiary services at academic health centres, and closing superfluous hospitals and other health facilities. America's pluralistic values and these proposals having serious political repercussions for health executives and elected officials often present serious barriers in their implementation. PMID- 12626022 TI - 'Best practice' development and transfer in the NHS: the importance of process as well as product knowledge. AB - A core prescription from the knowledge management movement is that the successful management of organizational knowledge will prevent firms from 'reinventing the wheel', in particular through the transfer of 'best practices'. Our findings challenge this logic. They suggest instead that knowledge is emergent and enacted in practice, and that normally those involved in a given practice have only a partial understanding of the overall practice. Generating knowledge about current practice is therefore a precursor to changing that practice. In this sense, knowledge transfer does not occur independently of or in sequence to knowledge generation, but instead the process of knowledge generation and its transfer are inexorably intertwined. Thus, rather than transferring 'product' knowledge about the new 'best practice' per se, our analysis suggests that it is more useful to transfer 'process' knowledge about effective ways to generate the knowledge of existing practice, which is the essential starting point for attempts to change that practice. PMID- 12626024 TI - Hospital efficiency and patient satisfaction. AB - The objective of this study was to investigate the relationship between efficiency and patient satisfaction for a sample of general, acute care hospitals in Ontario, Canada. A measure of patient satisfaction at the hospital level was constructed using data from a province-wide survey of patients in mid-1999. A measure of efficiency was constructed using data from a cost model used by the Ontario Ministry of Health, the primary funder of hospitals in Ontario. In accordance with previous studies, the model also included measures of hospital size, teaching status and rural location. Based on the results of this study, at a 95% confidence level, there does appear to be evidence to suggest that an inverse relationship between hospital efficiency and patient satisfaction exists. However, the magnitude of the effect appears to be small. Hospital size and teaching status also appear to affect satisfaction, with lower satisfaction scores reported among non-teaching and larger hospitals. This study did not find any evidence to suggest that hospital location (rural versus urban) or religious affiliation contributed to reports of patient satisfaction in any way not explained by the other measures included in the study. The findings imply that low patient satisfaction cannot be explained by excessive management concentration on efficiency. Managers should analyse some of the underlying causes of patient dissatisfaction before reconfiguring resources. It may be beneficial in larger hospitals to study the aspects of care that patients have reported they prefer in small hospitals. PMID- 12626025 TI - A theoretical model for the optimal allocation of health resources in Greece. AB - This paper presents a theoretical model that enables us to rate the efficiency of health resources in different regions. The model is based on the theory of production and refers to two specific regions. The hypotheses that determine the relationship between health care resources and their indicators are examined as well as the necessary conditions for the optimal regional allocation of these resources. PMID- 12626026 TI - Utilization of general practitioner services in New Zealand and its relationship with income, ethnicity and government subsidy. AB - The New Zealand primary care system involves user part-charges for general practitioner (GP) care. This study explores the relationship between the use of GP services in New Zealand and government subsidies, income and ethnicity. The study used the 1996/97 New Zealand Health Survey dataset, a nationally representative household survey of health status and health service utilization. Multivariate logistic regression models were used to estimate the likelihood of visiting the GP at least once in 12 months, and the likelihood of frequent visits (at least six visits in 12 months). Adjustment was made for health need (as indicated by global self-reported health status and a number of specific disease and health-related behaviour measures), together with a range of other relevant demographic and socio-economic variables. Following these adjustments, low-income groups and Maori were found to be significantly less likely to visit the GP at least once in the year, but there were no significant differences across income or ethnic groups in the likelihood of frequent visits. These results suggest that the system of low-income targeted government subsidies reduces, but does not fully compensate for the barrier posed by doctors' fees. Moreover, there appear to be barriers for indigenous groups, in addition to income. PMID- 12626027 TI - The role of regional planning and management strategies in the transformation of the healthcare system. AB - Drawing on a case study, this article questions the role of planning and management strategies in the process of transforming a regional public healthcare system that involves a number of organizations and is characterized by fluidity in its functions and division of power. It examines the efficacy of the Regional Plan for the Organization of Health Services (PROS) in reforming the mental health sector in a health and social service district in Quebec, in terms of integrated regional management of mental healthcare and activities at the local level. The regional planning procedure involves a major transformation in management of the mental health system, organizational roles and clinical and professional practices. Our assessment of PROS highlights the importance of taking into account the context of implementation and the instrumental value of planning, before judging its efficacy. To transform a complex healthcare system at the regional and local level, the study suggests a revised conception of the main roles played by planning and of the process shaping its implementation. Our study concludes in favour of developing management strategies at the operational, clinical and professional levels, and integrating them as a planning aid that allows a more corporate and matrix-based system to be set up. PMID- 12626028 TI - Intrauterine growth of normal thyroid. AB - In this study we describe fetal thyroid growth during gestation and establish normal reference values using a simple linear ultrasound measurement of the thyroid. A total of 1180 normal singleton pregnancies, with no known risk factors for thyroid disorders, were enrolled from 12 to 39 weeks of gestation. The thyroid antero-posterior diameter was measured on a transverse axial plane through the fetal neck. The best fit regression was a power equation: thyroid diameter = 0.2412 weeks(1.0278) (r(2) = 0.55). The percentiles smoothed curves were calculated for each week. In conclusion, the results of the present study support previous findings that the fetal thyroid grows between 12 and 39 weeks of gestation with a steepest increase during the second trimester, that is when the fetal thyroid becomes functionally active. The normal ranges of this simple index of thyroid growth can be useful both as screening and for the clinical evaluation of pregnant patients with thyroid disorders. PMID- 12626029 TI - Current concepts in aesthetic endocrinology. AB - The extragenital effects of ovarian steroids are relevant to the metabolism of skin and hair, the changes in body composition and the alterations of the subcutaneous fat distribution throughout life. When ovarian steroids become deficient or are produced in excess, different problems may arise in these tissues and some of these problems, i.e., obesity and cellulite, display gender specific components. Therefore, a new field in endocrine research known as aesthetic endocrinology is gaining more interest. Because sex steroids are small molecules they can be transported into the skin by topical application when properly formulated. This possibility is used in aesthetic endocrinology in order to achieve local effects but to avoid systemic reactions. After reviewing the current data it collectively seems legitimate to recommend estrogens, either orally or topically, in order to counteract the aging of the skin after menopause. Although a reconstitution of juvenile skin cannot be achieved through this method, a slowing in the skin aging process seems a reasonable expectation. In contrast, the successful treatment of hair loss in women is only confirmed for the application of the non-hormonal compound minoxidil. Apart from the difficult problem of hirsutism, acne and changes in body composition offer promising therapeutical options for endocrinological methods. PMID- 12626031 TI - The role of plasma renin activity in distinguishing patients with polycystic ovary syndrome (PCOS) from oligomenorrheic patients without PCOS. AB - All the components of the renin-angiotensin system have been identified in the human ovary. Renin plays a major role in folliculogenesis, and possibly in follicular atresia. Polycystic ovary syndrome (PCOS) is characterized by early follicular atresia. We studied whether assessment of plasma renin activity would be useful for diagnosing PCOS and distinguishing PCOS and non-PCOS in oligomenorrheic patients. Patients were divided into three groups: PCOS group (group 1, n = 40), non-PCOS oligomenorrheic group (group 2, n = 30) and ovulatory control group (group 3, n = 30). Plasma renin activity was determined in the early follicular phase of the menstrual cycle. Baseline serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin, insulin, aldosterone and androgens (total testosterone, free testosterone, androstenedione, dehydroepiandrosterone sulfate and 17alpha hydroxyprogesterone) were determined in all groups. The mean LH level was lower (4.94 +/- 4.65 mIU/ml) in control patients than in PCOS patients (12.9 +/- 1.75 mIU/ml) and non-PCOS oligomenorrheic patients (10.8 +/- 1.65 mIU/ml). There was no statistically significant difference between the groups regarding FSH levels. The mean plasma renin activity was 3.47 +/- 0.29 ng/ml in the PCOS group. The mean plasma renin activity was 1.59 +/- 0.21 ng/ml the non-PCOS oligomenorrheic group (statistically significiant differrence). There was no statistically significant difference between the non-PCOS oligomenorrheic (1.59 +/- 0.21 ng/ml) and control groups (1.2 +/- 0.16 ng/ml). Use of plasma renin activity alone as a diagnostic marker in PCOS is not useful, because the clinical findings of PCOS are heterogeneous and the pathogenesis is unclear. However, it can be used together with other variables to diagnose women with PCOS and to distinguish non-PCOS oligomenorrheic women from those with PCOS. PMID- 12626030 TI - Preliminary indication of a high prevalence of polycystic ovary syndrome in indigenous Australian women. AB - We aimed to estimate the likely prevalence of polycystic ovary syndrome (PCOS) in indigenous Australian women in a cross-sectional survey based on structured interviews. In the Kimberley region of Western Australia and south-western Victoria, between February and June 1999, women over the age of 18 available for interview and willing to participate were included. Menstrual status, anthropometric measures, fasting blood biochemistry and hirsutism were assessed. A total of 38 premenopausal women completed the survey. Of these, 50% had regular menstrual cycles of normal duration and no hirsutism; seven had oligomenorrhea and hirsutism/hyperandrogenemia; and of the nine women with hirsutism and regular menses, three had hyperandrogenemia. Thus, possibly ten women fulfilled the criteria of PCOS. This unique data set suggests a high prevalence of PCOS in indigenous Australian women. Further research to document this formally is warranted, in view of the potential adverse effects of PCOS on reproduction and metabolism. PMID- 12626032 TI - The pathophysiological influence of leptin and the tumor necrosis factor system on maternal insulin resistance: negative correlation with anthropometric parameters of neonates in gestational diabetes. AB - The contribution of the tumor necrosis factor (TNF) system and leptin was studied in insulin resistance and neonatal development during the course of normal pregnancy and gestational diabetes mellitus (GDM). Thirty patients with GDM and their neonates (n = 30), 35 healthy pregnant women (15 in the first, nine in the second and 11 in the third trimester) and their neonates (n = 20), and 25 healthy matched non-pregnant women participated in the study. Significantly elevated levels of maternal TNF-alpha, sTNF receptor (R)-1 and R-2, leptin (detected by enzyme-linked immunosorbent assay) and fasting C-peptide (measured by radioimmunossay and raised body mass index (BMI) were found in GDM patients and in the third trimester of normal pregnancies. TNF-alpha, sTNFR-2, C-peptide, leptin concentrations and BMI positively correlated with each other in GDM. An inverse relationship between the body length, head circumference and body weight of the newborns, and maternal TNF-alpha, leptin and C-peptide concentrations was shown in GDM. In healthy pregnancies the maternal serum leptin level was in a negative linear correlation with the head circumference of the newborns. In conclusion, increased TNF-alpha and leptin levels may contribute to insulin resistance in GDM and in the third trimester of normal pregnancy and may negatively influence the anthropometric parameters of the newborns. PMID- 12626033 TI - Effect of different hormone replacement therapy regimens on circadian blood pressure profile and active renin in postmenopausal women. AB - Hormone replacement therapy (HRT) was considered as main prevention of cardiovascular disease (CVD) in postmenopausal women. Mechanisms of vasoprotective effect of this treatment are complex. However, recent data give rise to some uncertainties about HRT benefits and risks. Little is known about the effects of oral and transdermal HRT regimens on the renin-angiotensin aldosterone system (RAS) and blood pressure (BP). This 3-month study comprised 28 menopausal women (age range 45-55 years) divided into two groups: Group 1: 12 normotensive women with natural occurrence of menopause receiving oral treatment with Climen (Schering) containing estradiol valerate and cyproterone acetate; Group 2: 16 normotensive women with surgically induced menopause receiving transdermal application of Climara (Schering) containing 17beta-estradiol. There were no significant differences in office BP before and after treatment with Climara or Climen. However, ambulatory monitoring showed a significant fall in systolic BP (day-time, night-time and total 24-h) when estradiol alone was used. A similar trend towards lower values of systolic BP that was significant only for the night-time BP was observed after treatment with Climen. There were no significant changes in diastolic BP after both treatment regimens. Heart rate (day-time and 24-h) was significantly lower after transdermal estradiol treatment. There was no significant change in active renin after both treatment regimens. The present study showed that both treatment regimens resulted in lower ambulatory BP in normotensive postmenopausal women with more notable reduction in night-time BP. Increase in nocturnal dipping may account in part for the beneficial cardiovascular effects of HRT including decreased end-organ damage. PMID- 12626034 TI - Hormone replacement therapy in women with a history of breast cancer. AB - Health care professionals in modern Western societies will meet an increasing number of women surviving breast cancer. How the menopause of these women should be treated is still an open question. Use of hormone replacement therapy (HRT) may, at least in theory, increase the risk for recurrence of cancer, but its categoric refusal is a double-edged sword because it also denies these women all the undisputable health benefits HRT provides. This refusal is not, however, supported by the observational data available so far on this question, because HRT has not increased the risk for breast cancer recurrence. In fact, it is well established that HRT abolishes hot flushes and improves significantly these patients' quality of life. At present, we have no effective nonhormonal alternatives for the control of vasomotor symptoms, and the efficacy of phytoestrogens in the treatment of menopausal symptoms is unproven. Selective estrogen receptor modulators (SERMs) which protect against osteoporosis and perhaps also against breast cancer, and which may have beneficial effects on the cardiovascular system, aggravate hot flushes and are therefore not useful, at least in the first postmenopausal years. In some countries, progestins are often prescribed for the control of such patients' vasomotor symptoms, but their safety has never been assessed in clinical trials, and in theory they can be harmful. Randomized clinical trials (RCT) on the use of HRT in breast cancer survivors are underway, but their completion will take years, and even these may be open to criticism. Tibolone may appear to be an appealing alternative for HRT, but it should also be studied with RCTs in this indication. At present, a patient with a history of breast cancer must be given balanced information as to the possible benefits and risks of HRT, and she herself must make the decision whether or not to start HRT. PMID- 12626035 TI - Effects of sex steroid hormones and menopause on serum leptin concentrations. AB - Leptin is a protein secreted by adipocytes; its circulating levels are correlated to fat mass and it acts on the hypothalamic centers regulating body weight. Leptin may also play an important role in regulating reproductive function. Indeed, ob/ob mice, lacking leptin due to a genetic mutation, are obese and infertile; administration of recombinant leptin to these animals reduces body weight and restores fertility. A sexual dimorphism in serum leptin levels has also been observed, with higher concentrations in women. Studies in vitro seem to indicate that estrogens stimulate leptin secretion, while in vivo studies are extremely discordant. In humans, several studies showed increased, unmodified and decreased leptin levels after the menopause. Furthermore, hormonal replacement therapy (HRT) after the menopause was reported to result in unmodified, increased or decreased leptin levels. It is likely that the effects of postmenopausal hypoestrogenism on leptin levels are masked by the postmenopausal changes in body composition. Indeed, after menopause, there is an increase in body weight, body mass index (BMI) and fat mass with a centralization of fat distribution. Administration of HRT may stop these changes and even restore a premenopausal pattern, leading then to decreased leptin levels. PMID- 12626036 TI - Molecular mechanisms contributing to the pathogenesis of endometriosis. PMID- 12626037 TI - Flow cytometry in human reproductive biology. AB - Flow cytometry (FC) is an analytical cytology technique which has been extensively used for decades. It has many advantages compared with other similar methods for the study of cell biology, even on a molecular basis. FC allows the cell-by-cell analysis of many optical or immunological features in the same sample, at the same time, and at a rate of thousands of cells per second, generating immense quantities of data and thus providing almost limitless information which is statistically robust due to the number of units studied. The aim of this review is to describe the contribution of FC to the study of physiological and pathological processes related to human reproduction, and to discuss how this technique has been used in research, as well as its clinical applications in this field. We have used some practical examples selected from the most relevant studies within a wide range of investigations published in the literature, and we have also drawn on our own experience of using flow cytometry to study different phenomena related to reproduction. It is conclued that FC is a useful instrument for basic investigation of gynecological issues, as well as for the study of male reproductive characteristics, either in research applications or directly for clinical diagnostic purposes. Future development of these techniques will permit further advances both in our knowledge and in the improvement of assisted reproduction techniques. PMID- 12626057 TI - Research for medical photographers: photographic measurement. AB - Many first degrees in photography or photography-related topics lack a thorough grounding in the photographic sciences. This in turn leads to difficulties for medical photographers who graduate from these courses, when it comes to designing photographic protocols for research projects that will utilize photography as a means of data collection. The concept of the image as subject analogue is introduced and means of visualizing the subject are outlined. The considerations for undertaking photometric, photogrammetric and time-based measurements are given. The variables that need to be controlled in each modality are highlighted and a bibliography is appended so that readers can discover for themselves working examples of the techniques described. PMID- 12626058 TI - Designing trigger pictures in context: the challenge of balance. AB - Experience, supported by research, has shown that people interpret pictures against a complex network of communication factors, including various socio cultural factors. In the process of developing interactive health learning materials for primary schools of less developed communities of South Africa, a set of prototype trigger pictures was pre-tested to determine its suitability for sexuality and HIV/AIDS education. The results suggest a link between socio cultural factors related to sexual behaviour among young people, and the way they 'read' these pictures. By way of an explanatory model it is recommended that media producers should seek a balance between learner factors, picture coding factors, and socio-cultural factors in the design of contextualized trigger pictures. PMID- 12626059 TI - The ethical context of clinical photography. AB - The unforseen consequences of unconsented clinical photography when images are required for publication is an issue that affects medical practitioners and clinical photographers. This paper discusses, a scenario based on a typical event, the nature of informed consent and its relationship to ethical theory and the development of codes of practice. It includes a critique of ethical theory and describes mapping or codification of moral behaviour and the relationship to protocols or codes of practice, and concludes with a call to improve understanding and expertise in the study of ethics and its context in medical illustration. PMID- 12626060 TI - Planning, designing and evaluating a departmental medical illustration website. AB - The Medical Illustration Department at West Glasgow Hospitals made a decision to construct a departmental website that would not only raise awareness of the department but also demonstrate the expertise in website construction the medical illustration staff had developed. Departmental staff and client base surveys were conducted to define what was required for a successful departmental website. Further investigation was conducted by surveying five existing medical illustration websites. Based on this initial data the website was constructed. A post-construction survey was conducted to assess suitability, usability and to highlight any potential problem areas. PMID- 12626061 TI - Masking unmasked. PMID- 12626062 TI - The preservation and destruction of archival material. PMID- 12626063 TI - Keratic precipitates in uncontrolled glaucoma. PMID- 12626064 TI - Biocomm 2002 - Bio-Communications Association, Orlando, February 22-23, 2002. PMID- 12626067 TI - Characterization of transgenic mice--a comparison of protocols for welfare evaluation and phenotype characterization of mice with a suggestion on a future certificate of instruction. AB - A thorough welfare evaluation performed as part of a general phenotype characterization for both transgenic and traditional mouse strains could not only contribute to the improvement of the welfare of laboratory animals, but could also be of benefit to scientists, laboratory veterinarians and the inspecting authorities. A literature review has been performed to identify and critically evaluate already existing protocols for phenotype and welfare characterization. There are several relevant schemes available, among others the SHIRPA method, the modified score sheet of Morton and Griffiths, the FRIMORFO phenotype characterization scheme and the behavioural phenotype schemes as described by Crawley. These protocols have been evaluated according to four goals: Their ability (1) to reveal any special needs or problems with a transgenic strain, (2) to cover the informational needs of the purchaser/user of the strain, (3) to refine the welfare of the transgenic animal model by identifying relevant humane endpoints, (4) to prevent the duplication of animal models that have already been developed. The protocols described are useful for characterizing the phenotype and judging welfare disturbances, however the total amount of information and the degree of detail varies considerably from one scheme to another. We present a proposal regarding the practical application of the various schemes that will secure proper treatment and the identification of humane endpoints. It is advocated that with every purchase of a particular strain, an instruction document should accompany the strain. This document needs to give detailed descriptions of the typical characteristics of the strain, as well as necessary actions concerning relevant treatment and humane endpoints. At the moment no such documents are required. The introduction of these types of documents will contribute to improvements in animal welfare as well as experimental results in laboratory animal experimentation. PMID- 12626068 TI - The refining influence of ethics committees on animal experimentation in Sweden. AB - Mandatory scrutiny of projects by animal ethics committees was introduced in Sweden in 1979. The present study investigated the minutes of meetings held between 1989 and 2000 at which consideration of applications for experimental work in animals resulted in requests for modification (n = 3607). 18.1% of the applications received were approved only after modifications. The majority of the changes requested may be classified as 'Refinement'. The most common requests were for improvement of project design, euthanasia method and housing and husbandry. There was a relative increase in modifications requested by the committees related to anaesthesia, choice of licensed supervisor and the need for licenses or informed consent from animal owners during the period investigated. There was a relative decrease in modifications related to euthanasia, housing and husbandry, and general endpoint assertions. The results suggest that the work of the committees may be perceived as an ongoing process, since several of the applications for which modification was requested were projects that had been approved on a previous occasion but were now up for renewal. In order to have maximal influence on the refinement of scientific protocols it is important that the scientists in the committees are continuously updated on developments in laboratory animal science. PMID- 12626069 TI - The Himalayan rabbit (Oryctolagus cuniculus L.): spontaneous incidences of endpoints from prenatal developmental toxicity studies. AB - The historical database represents the spontaneous, i.e. normal changes of characters, in laboratory animals, and is a must for the assessment of toxic effects of compounds. For the Himalayan rabbit (CHBB:HM, SPF) background data were collected from the control groups of regulatory studies between 1968-1999 in the Biberach laboratory of Boehringer Ingelheim (Germany). In the present study these data were compared with the background data from the years 1974-1984, with those from the Ingelheim laboratory, Germany (1977-1984) and those from the breeding colony at Kawanishi, Japan (1971-1991, 1972-1984). Fertility in the Biberach laboratory was relatively constant through the years, due to the strict breeding system. Litter parameters (corpora lutea, implantations and viable fetuses), including fetal sex distribution and fetal weight, were constant in the Biberach laboratory through the years and compared well with the data from Ingelheim and Kawanishi. From these three laboratories the results of spontaneous changes in fetal morphology above 0.1% incidence suggest a pattern of deviations from the Bauplan (fundamental body plan). These comprise synosteosis of sternebrae, lumbar (additional) ribs, hypoplasia of 12th rib, status of ossification, aplasia of gall bladder, hypoplasia of gall bladder, incompletely subdivided lung, dilated ventricle of heart, deviations at the A. carotis and at the aortic arch and flexure of paw. The absence of a time-dependent statistical trend indicates that the spontaneous change of endpoints remained constant through the time axis. Comparison with the spontaneously changed fetal morphology of the other strains (NZW, JW, SF rabbits) does not provide support for their being a totally strain-specific pattern, and this pattern may be closer to the species than specific to a single strain. The firm patterns of deviations in fetal morphology provide suitable and valuable markers for assessing toxic effects on ontogenesis. According to our experience these morphological endpoints react very sensitively to teratogenic compounds, which is demonstrated by increased incidences. Our knowledge of evolution of organisms and of mechanisms leading to morphological deviations points to a strict selection of phenotypes. For laboratory animals the decisive selection factors are defined by man. These are nutritive factors, environmental conditions, the mating system, handling and treatment, and experimental design. All these factors have to be held constant through the years, which is an absolute pre-condition for the application of valid historical data. The Himalayan rabbit is highly suitable for use in developmental toxicity studies due to the stability of reproductive data within the same laboratory over a period of more than 30 years, due to the similarity of reproductive data among three different laboratories, and due to an absence of relevant differences to the reproductive data of other rabbit strains. PMID- 12626070 TI - Lymph node topography of the head and neck in New Zealand White rabbits. AB - Investigations of the lymphogenic metastatic spread of VX2 carcinomas in New Zealand White rabbits require an exact knowledge of the topography of cervical and facial lymph nodes. The topography of neck lymph nodes was evaluated from 16 rabbits macroscopically, histologically and by lymphographic investigations, and the possibility of their surgical removal (neck dissection) was examined. The upper aerodigestive tract and the ear of New Zealand White rabbits drain via four consistent groups of 12-18 lymph nodes. Except for the paratracheal lymph node, they are all easily accessible to surgery. The data presented in this study encourage the use of induced VX2 carcinomas in New Zealand White rabbits as an animal model to study the lymphogenic metastatic spread of squamous cell carcinomas of the head and neck. Such investigations could lead to an improvement of surgical and pharmaceutical treatment of this tumour entity. PMID- 12626071 TI - The effects of different rack systems on the breeding performance of DBA/2 mice. AB - Housing systems for laboratory animals have been developed over a long time. Micro-environmental systems such as positive, individually ventilated caging systems and forced-air-ventilated systems are increasingly used by many researchers to reduce cross contamination between cages. There have been many investigations of the impact of these systems on the health of animals, the light intensity, the relative humidity and temperature of cages, the concentration of ammonia and CO(2), and other factors in the cages. The aim of the present study was to compare the effects of different rack systems and to understand the influence of environmental enrichment on the breeding performance of mice. Sixty DBA/2 breeding pairs were used for this experiment. Animals were kept in three rack systems: a ventilated cabinet, a normal open rack and an individually ventilated cage rack (IVC rack) with enriched or non-enriched type II elongated Makrolon cages. Reproduction performance was recorded from 10 to 40 weeks of age. In all three rack systems there was a similar breeding index (pups/dam/week) in non-enriched groups during the long-term breeding period, but the coefficients of variation in the IVC rack were higher for most parameters. This type of enrichment seems to lead to a decrease in the number of pups born, especially in the IVC group. However, there was no significant difference in breeding index (young weaned/female/week). PMID- 12626072 TI - Higher heart rate of laboratory mice housed individually vs in pairs. AB - Many studies have shown that housing mice individually over a long period significantly alters their physiology, but in most cases measurement has required human interference and restraint for sampling. Using a radio-telemetry system with implantable transmitters, we recorded heart rate (HR), motor activity (ACT) and body temperature (BT) of freely moving male mice (NMRI) housed either individually or in pairs with an ovarectomized female. Data for each parameter were collected at 5 min intervals for two consecutive 24 h periods. Even after several weeks of habituation to the social conditions, HR was increased in mice housed individually compared with mice housed in pairs, although their measured ACT did not differ. Additionally, BT tended to be reduced in individually-housed mice. When the data were analysed according to different ACT levels, HR was increased in individually-housed mice during phases of low and high, but not intermediate, motor activity. Furthermore, individually-housed mice had more, but shorter, resting bouts, indicating disruption of the normal circadian sleep pattern. Enhanced HR in individually-housed mice does not necessarily indicate stress, but might be an important physiological indicator of discomfort. The fact that individual housing alters basic physiological parameters in laboratory mice highlights the need to control for housing-dependent variation, especially in experiments that are sensitive to changes in these parameters. PMID- 12626073 TI - Subsensitivity to beta-adrenergic stimulation in atria from rats infested with Syphacia sp. AB - Syphacia sp. is a common intestinal parasite in conventionally-housed laboratory rodents. Although gross lesions are rare in oxyuriasis, it is possible that more subtle changes may develop, which may affect research results. In this study, we analysed the responsiveness to beta-adrenergic stimulation by isoproterenol (ISO) of left atria isolated from Syphacia-infested (SYPH) and control, non-infested adult male Wistar rats (CONT). In the non-infested animals, ISO pD(2) was not significantly changed by ivermectin treatment. Whereas the maximal inotropic response to ISO was not significantly affected, the pD(2) value was decreased in SYPH (7.61 +/- 0.09, n = 7, vs 8.21 +/- 0.25 in CONT, n = 5, P < 0.05), indicating lower sensitivity to beta-adrenergic stimulation. This change was similar to that caused by a classic stressor, namely repeated immobilization, in non-infested rats (IMMO). In this group, ISO pD(2) was 7.62 +/- 0.14, n = 6 (P < 0.05 with relation to CONT). The results indicate that infestation with Syphacia sp. is as effective as immobilization at diminishing cardiac reactivity to beta adrenergic stimulation. It is thus possible that oxyuriasis may affect the response of other tissues to physiological modulators. PMID- 12626074 TI - A simple procedure to perform intravenous injections in the Mongolian gerbil (Meriones unguiculatus). AB - We describe a simple and feasible procedure for performing intravenous administration of substances in the gerbil. Under light anaesthesia, animals were held in dorsal recumbency and a very small incision of skin, parallel to the femoral vein on the internal side of the thigh, was made. The vein is easily accessible via thin skin incision. An insulin syringe and a 30 G needle were used for the injection. This is an easy and quick method, which, with appropriate anaesthesia, allows rapid recovery. PMID- 12626075 TI - The effect of body weight on the degree of blood velocity profile skewness in the aortic annulus in domestic pigs. AB - We investigated the blood velocity profile in the aortic annulus (AA) in two groups of domestic pigs using epicardial Doppler echocardiography. The velocity profile skewness in terms of max/mean TVI (the ratio of maximal to cross sectional mean time-velocity integral along the diameter) was 1.107 +/- 0.01 in the small pigs (n = 10; body weight: 24.6 +/- 0.8 kg) and 1.216 +/- 0.026 in the large pigs (n = 8; body weight: 50.6 +/- 2.5 kg) (P = 0.002). The velocity profile in the AA is more skewed in large animals than in small animals and the skewness in the larger animals is similar to that in normal adult humans. This study shows the importance of choosing animals of sufficient size if flow method investigations are to be performed. This is particularly important for ultrasound Doppler investigations based on a limited sample of velocities across the flow channel. PMID- 12626077 TI - Toward optimal health: the experts discuss osteoarthritis in women [interviewed by Jodi Godfrey Meisler]. PMID- 12626078 TI - External occlusive devices for management of female urinary incontinence. AB - OBJECTIVES: Urinary incontinence affects 11 million women in the United States and is most common in older women. There are several different forms of urinary incontinence, which can be distinguished by their associated pathophysiologic conditions and signs. Genuine stress incontinence is the instantaneous leakage of urine in response to raised intraabdominal pressure associated with such activities as lifting, sneezing, and coughing. Risk factors for development of this form of incontinence include vaginal delivery, vaginal surgery, inadequate estrogen levels, and advanced age. Surgery, the most effective curative treatment for stress incontinence, is invasive and expensive and can lead to impairment of normal urinary tract functions. Other management options include pelvic floor rehabilitation by means of exercise, pharmacotherapy, intravaginal and intraurethral devices, absorbent products, and external occlusive devices. METHODS: A review of publications on the effectiveness of available external occlusive devices is presented, with special emphasis on a new, single-use, disposable urethral cap device which offers women with stress incontinence an over-the-counter product that enables them to independently manage their incontinence according to their individual needs. RESULTS: Non-surgical methods of managing urinary incontinence are available, with newer methods providing a greater range of choices for women. CONCLUSIONS: External occlusive devices can be effectively used by women with urinary incontinence who will not or cannot have surgical correction of the problem. PMID- 12626079 TI - Study of axial bone mineral density in postmenopausal women with diffuse idiopathic skeletal hyperostosis related to type 2 diabetes mellitus. AB - OBJECTIVE: The association of bone mineral density (BMD) with diffuse idiopathic skeletal hyperostosis (DISH) related to diabetes mellitus was studied. METHODS: We measured BMD and elevated known determinants of BMD (bone markers) in 35 patients with DISH-related type 2 diabetes mellitus, 47 type 2 female diabetics, and 52 female controls with no systemic disease and no drug administration. All subjects were matched for age and body mass index (BMI). All subjects were in the postmenopausal period. RESULTS: Among subjects, BMD values were significantly higher in DISH patients than in diabetics and controls (p < 0.05). In addition, the duration of diabetes mellitus was longer in DISH patients and significantly correlated with total hip BMD in DISH patients (p < 0.05). CONCLUSIONS: Patients with DISH have higher BMD, and increased BMD probably results from hyperostosis of the axial skeleton, which may cause decreased fracture risk. PMID- 12626080 TI - Hormone replacement therapy counseling: prevalence and predictors. AB - OBJECTIVES: To evaluate the prevalence of hormone replacement therapy (HRT) counseling and to investigate the relationship between patients' cultural background and healthcare professionals' counseling practices. HRT is the principal preventive intervention for reducing risk of menopause-related pathologic conditions. This study assessed self-reports of HRT counseling among women receiving care in an urban community hospital. METHODS: A descriptive self reported survey study of 134 women aged 45-65 was conducted in the medical clinic at an urban community teaching hospital in a multicultural area in New York. Questionnaires asked about age, ethnicity, HRT use, contraindications, health professional counseling, and barriers to HRT counseling. Possible predictors of HRT counseling were analyzed with logistic regression models. Categorical variables were compared with chi-square statistics. RESULTS: Overall prevalence of HRT counseling was 25%. Prevalence of HRT use was 19%; among those using HRT, only 12.7% reported receiving HRT counseling. Physicians were more likely to discuss HRT than other healthcare professionals. Over 90% of the patients reported a desire for HRT counseling. CONCLUSIONS: The low rates of HRT counseling suggest physicians have limited daily experience with HRT counseling practices. Public health efforts should focus on increasing physician and general population awareness to promote HRT counseling strategies and ensure public access to HRT information. PMID- 12626081 TI - Aging, health, and depressive symptoms: are women and men different? AB - OBJECTIVE: This study investigated whether age and self-reported health status comparisons affected reports of depressive symptoms differently for older women and men. METHODS: Cross-sectional survey design conducted in a large health maintenance organization (HMO) that provides comprehensive, prepaid benefits to its members. Study subjects were enrollees who were 65 years of age or older. The dependent measure was their reports of depressive symptoms as measured using the Center for Epidemiologic Studies-Depression Scale (CES-D). The explanatory variables were patients' gender, age, self-reported health status comparisons, living status, and physical impairments. RESULTS: The difference in the proportion of females reporting high depressive symptoms (CES-D>or=16) was 3.8% between the 75-84 year age group and the youngest female group (26.9% vs. 23.1%, respectively). The difference for males was 13.7% (26.5% vs. 12.8%, respectively) (OR(5) = 0.44, 95% CI(5) = 0.27, 0.73). Similar findings occurred among females and males who self-reported that they were in poor health (OR(5) = 0.20, 95% CI(5) = 0.06, 0.72). Aging and self-reported comparisons affected females and males similarly when high depressive symptoms were examined using a higher cut point (CES-D>or=21), although females were more than 50% more likely to report symptoms than males. CONCLUSIONS: As females aged and self-reported health status comparisons declined, their depressive symptoms increased steadily, but at a lower rate than males' depressive symptoms. Older persons in fair or poor self reported health should especially be screened for increasing depressive symptoms. Although females generally report greater risk of high depressive symptoms, it is still a concern for males as they age and their health declines. PMID- 12626083 TI - Observations from the CDC. REACH 2010 coalitions: reaching for ways to prevent cardiovascular disease and diabetes. PMID- 12626084 TI - Toward optimal health: the experts discuss the issues of aging. Interview by Jodi Godfrey Meisler. PMID- 12626085 TI - Lambert-eaton myasthenic syndrome: a case report and review of the literature. AB - Lambert-Eaton myasthenic syndrome (LEMS) is a type of paraneoplastic syndrome that may initially manifest with weakness and gait abnormalities. These symptoms may precede the diagnosis of malignancy by months or years, and morbidity and mortality may be significantly affected by early detection of the malignancy. A case report and review of the diagnosis, management, and treatment of these syndromes are presented, with particular emphasis on the rehabilitation management of these patients, often overlooked in medical treatment. PMID- 12626086 TI - Vaginal estrogen preparations: a review of safety and efficacy for vaginal atrophy. AB - BACKGROUND: A theoretical advantage of local (i.e., vaginal) therapy of genitourinary symptoms could be avoidance of systemic adverse effects. Review of efficacy and adverse effects of commonly prescribed vaginal estrogens is of great clinical relevance. METHODS: A Medline (1966-present) search was performed for randomized controlled trials involving vaginal estrogens. Reference lists of papers were reviewed for additional references. RESULTS: Twenty-two references were randomized controlled trials of vaginal estrogens used by postmenopausal women with signs or symptoms of vaginal atrophy. Subject numbers ranged from 20 to 251. Duration ranged from 2 weeks to 1 year. Different preparations and schedules were used across the trials. All treatments alleviated signs and symptoms of atrophic vaginitis, regardless of whether objective signs of atrophy were required for study entry. Data for urinary symptoms was conflicting; the ring may prevent recurrent urinary tract infections (UTIs). The trials with endometrial scrutiny were less than one year and had mixed results. Nonhormonal lubricant is effective in improving some atrophic signs and symptoms. All preparations were associated with vaginal irritation. Bleeding with vaginal estradiol tablets may be less than that with CEE cream. Vaginal tablets or rings were preferred over other preparations. There were no serious adverse events reported. There was occasional expulsion of estradiol ring in the setting of prior hysterectomy. CONCLUSIONS: All preparations are effective in decreasing signs and symptoms of vaginal atrophy, but they differ slightly in their adverse event profiles. Long-term safety of the preparations is best established for estradiol tablets (1 year), but is lacking for all preparations. PMID- 12626087 TI - Urinary incontinence in working women: a comparison study. AB - OBJECTIVES: To compare the findings of two surveys concerning the nature of urinary incontinence and management strategies used by full-time employed working women. METHODS: The first study was conducted in an urban academic setting with a survey distributed to 2000 women. The second study was conducted with 500 women in a rural pottery manufacturing facility. RESULTS: Twenty-one percent of the women working in an academic setting (group A) and 29% (p = 0.002) of the women working in a manufacturing setting (group B) reported being incontinent of urine at least monthly. More women in group B reported antecedents mixed incontinence (i.e., antecedent to both stress and urge incontinence). More women in group B also used panty liners to manage urine loss (p = 0.003), whereas more women in group A used voiding schedules (p = 0.008) and pelvic muscle exercises (p = 0.04). More women in group A reported that they did not know if their incontinence could improve (40% vs. 1%, p = 0.00). The majority in both groups said that getting treatment was of no or little importance, yet, overwhelmingly, most women (group A 81%; group B 86%) wanted more information about incontinence. CONCLUSIONS: Incontinence is a prevalent condition in working women. Discrepancies exist in the nature of the urine loss and strategies used to manage incontinence. Treatment also differs for those women who report urine loss to healthcare providers. Most women want to learn more about incontinence. Secondary prevention strategies need to be tailored and tested to meet symptoms and concerns for women in different work settings. PMID- 12626088 TI - The risk for infant mortality among adolescent childbearing groups. AB - OBJECTIVE: To evaluate risk disparities and risk factors for infant mortality among adolescent childbearing age groups. METHODS: We combined the 1995 and 1996 comprehensive U.S. birth cohorts provided by the National Center for Heath Statistics. Our analysis included 777,762 singleton, first births to women aged 12-19 years linked to 4631 infant deaths. We used both bivariate comparisons and multivariable logistic regression for our analysis, with infant mortality as our main outcome measure. RESULTS: Rates of infant mortality are substantially higher for < or =15-year-olds (8.1/1000 live births) compared with 16-17-year-olds (6.3/1000 live births) and 18-19-year-olds (5.4/1000 live births). Even after adjusting for risk factors associated with poor outcomes, including alcohol use, tobacco use, and prenatal care use, the risk for infant mortality was 1.6 (95% confidence interval [95% CI] 1.4, 1.7) times greater for infants of mothers < or =15 years old as compared with those mothers 18-19 years old. In the < or =15 year-old group, 62% of fathers were not reported on the child's birth certificate. Not reporting the father was associated with a 24% increased risk for infant mortality after adjusting for maternal and infant risk factors. CONCLUSIONS: Childbearing in < or =15-year-olds is associated with a substantial increased risk for infant mortality compared with childbearing in older adolescence. This study suggests that not reporting the father on a birth certificate is a potential risk marker. Risk differences among adolescent age groups may be important to consider when creating tailored intervention and prevention strategies. PMID- 12626089 TI - Religious beliefs and breast cancer screening. AB - BACKGROUND: The mechanisms underlying a reported tendency for women who hold strong religious beliefs to seek medical help at more advanced stages of breast cancer are unknown. This study investigates further the effect of religious beliefs with other variables on breast cancer screening and the intended presentation of a self-discovered breast lump. METHODS: The study included 682 eastern North Carolina women aged 40 and over who were interviewed in their homes about religious and other beliefs about breast cancer, screening, and intended actions with a self-discovered breast lump. RESULTS: Principal components factor analysis results suggested that a majority of women believe that God works through doctors to cure breast cancer. We labeled this dimension "religious intervention with treatment." A minority believed that medical treatment was unnecessary because only God could cure breast cancer. We labeled this dimension "religious intervention in place of treatment." The first dimension correlated with self-reported mammography but not clinical breast examination or women's intention to delay presentation of a self-discovered breast lump. The second dimension, significantly more common in African American women who were less educated and older, correlated strongly with the intention to delay presentation of a self-discovered breast lump. CONCLUSIONS: Belief in "religious intervention in place of treatment" may help to explain why African American women delay presentation of palpable breast lumps, contributing to advanced-stage cancer diagnosis. We suggest that clinicians and clergy work together within the context of religious beliefs to enhance early detection and survival from breast cancer. PMID- 12626091 TI - Carbon sequestration in Synechococcus Sp.: from molecular machines to hierarchical modeling. AB - The U.S. Department of Energy recently announced the first five grants for the Genomes to Life (GTL) Program. The goal of this program is to "achieve the most far-reaching of all biological goals: a fundamental, comprehensive, and systematic understanding of life." While more information about the program can be found at the GTL website (www.doegenomestolife.org), this paper provides an overview of one of the five GTL projects funded, "Carbon Sequestration in Synechococcus Sp.: From Molecular Machines to Hierarchical Modeling." This project is a combined experimental and computational effort emphasizing developing, prototyping, and applying new computational tools and methods to elucidate the biochemical mechanisms of the carbon sequestration of Synechococcus Sp., an abundant marine cyanobacteria known to play an important role in the global carbon cycle. Understanding, predicting, and perhaps manipulating carbon fixation in the oceans has long been a major focus of biological oceanography and has more recently been of interest to a broader audience of scientists and policy makers. It is clear that the oceanic sinks and sources of CO(2) are important terms in the global environmental response to anthropogenic atmospheric inputs of CO(2) and that oceanic microorganisms play a key role in this response. However, the relationship between this global phenomenon and the biochemical mechanisms of carbon fixation in these microorganisms is poorly understood. The project includes five subprojects: an experimental investigation, three computational biology efforts, and a fifth which deals with addressing computational infrastructure challenges of relevance to this project and the Genomes to Life program as a whole. Our experimental effort is designed to provide biology and data to drive the computational efforts and includes significant investment in developing new experimental methods for uncovering protein partners, characterizing protein complexes, identifying new binding domains. We will also develop and apply new data measurement and statistical methods for analyzing microarray experiments. Our computational efforts include coupling molecular simulation methods with knowledge discovery from diverse biological data sets for high-throughput discovery and characterization of protein-protein complexes and developing a set of novel capabilities for inference of regulatory pathways in microbial genomes across multiple sources of information through the integration of computational and experimental technologies. These capabilities will be applied to Synechococcus regulatory pathways to characterize their interaction map and identify component proteins in these pathways. We will also investigate methods for combining experimental and computational results with visualization and natural language tools to accelerate discovery of regulatory pathways. Furthermore, given that the ultimate goal of this effort is to develop a systems level of understanding of how the Synechococcus genome affects carbon fixation at the global scale, we will develop and apply a set of tools for capturing the carbon fixation behavior of complex of Synechococcus at different levels of resolution. Finally, because the explosion of data being produced by high throughput experiments requires data analysis and models which are more computationally complex, more heterogeneous, and require coupling to ever increasing amounts of experimentally obtained data in varying formats, we have also established a companion computational infrastructure to support this effort as well as the Genomes to Life program as a whole. PMID- 12626092 TI - Analysis of the genetic potential and gene expression of microbial communities involved in the in situ bioremediation of uranium and harvesting electrical energy from organic matter. AB - The proposed research will investigate two microbial communities that are of direct relevance to Department of Energy interests. One is the microbial community associated with the in situ bioremediation of uranium-contaminated groundwater. The second is a microbial community that harvests energy from waste organic matter in the form of electricity. These studies will address DOE needs for (1) remediation of metals and radionuclides at DOE sites and (2) the development of cleaner forms of energy and biomass conversion to energy. Our previous studies have demonstrated that the microbial communities involved in uranium bioremediation and energy harvesting are both dominated by microorganisms in the family Geobacteraceae and that the organisms in this family are responsible for uranium bioremediation and electron transfer to electrodes. The initial objectives of this study are to (1) describe the genetic potential of the Geobacteraceae that predominate in the environments of interest; (2) identify conserved patterns of gene expression within the Geobacteraceae family in response to a range of environmental conditions; (3) begin to identify mechanisms controlling the expression of key genes related to survival, growth, and activity in subsurface environments and on electrodes; and (4) use the results from subobjectives 1-3 to develop a conceptual model for predicting gene expression of Geobacteraceae in the environments of interest. This will serve as the basis for a subsequent simulation model of the growth and activity of Geobacteraceae in the subsurface and on electrodes. PMID- 12626093 TI - H. influenzae Consortium: integrative study of H. influenzae-human interactions. AB - Developments in high-throughput analysis tools coupled with integrative computational techniques have enabled biological studies to reach new levels. The ability to correlate large volumes of diverse data types into cohesive models of organism function has spawned a new systematic approach to biological investigation. The creation of a new consortium has been proposed to investigate a single organism utilizing these comprehensive approaches. The Haemophilus influenzae Consortium (HIC) would be comprised of five laboratories, each providing separate and complementary areas of expertise in the study of Haemophilus influenzae (HI). The 5-year study proposes to develop coherent models of HI, both as a stand-alone organism, and more importantly, as a human pathogen. Studies in growth condition specificity followed by genomic, metabolic, and proteomic experimentation will be combined and integrated through computational and experimental analyses to form dynamic and predictive models of HI and its responses. Data from the HIC will allow greater understanding of cellular behavior, pathogen-host interactions, bacterial infection, and provide future scientific endeavors with a template for studies of other pathogens. PMID- 12626094 TI - Target selection for structural genomics: a single genome approach. AB - We describe our strategy for selecting targets for protein structure determination in context of structural genomics of a single genome. In the course of target selection, we have studied two of the smallest microbial genomes, Mycoplasma genitalium and Mycoplasma pneumoniae. To our surprise, we found that only 71 Mycoplasma genes or their orthologues can be considered as easy targets for high-throughput structural studies--far fewer than expected. We discuss the methods and criteria used for target selection and the reasons explaining rarity of easy targets. First, despite the common opinion that protein folds can be predicted for only 30-50% of genes, the number of "truly unknown" structures is less than one-third. Second, due to the different codon usage, two thirds of Mycoplasma proteins cannot be directly expressed in E. coli in high-throughput manner and require substitution by their homologues from other organisms. Third, membrane or large multi-domain proteins are difficult targets because of solubility and size issues and often require identification and structure determination of protein domains. Finally, we propose different approaches to address the difficult targets. PMID- 12626095 TI - Improved postimplantation development of rabbit nuclear transfer embryos by activation with inositol 1,4,5-trisphosphate. AB - Cloned rabbit embryos are characterized by their extremely poor postimplantation development, despite their high survivability until the blastocyst stage in vitro. This study examined whether the developmental failure of cloned rabbit embryos in vivo can be overcome by technical improvements to the activation protocol. Freshly collected cumulus cells were transferred into enucleated oocytes by intracytoplasmic injection. One to two hours later, the oocytes were activated by electroporation with Ca(2+) or inositol 1,4,5-trisphosphate (IP3), which is known to induce repeated rises in intracellular Ca(2+), as in normal fertilization. After transfer of embryos at the two- to four-cell stages, well defined implantation sites with remnant fetal tissue were observed at term (day 28) only in the IP3-stimulation groups (0.9% and 5.8% per transferred embryo for single and triple stimulation groups, respectively). When some recipients in the same group were examined at days 16-20, a viable cloned fetus (day 19) with normal organogenesis was obtained. These findings clearly demonstrate that the oocyte activation protocol using IP3 enhances the postimplantation development of nuclear-transferred rabbit embryos. PMID- 12626096 TI - Chromosomal abnormalities: a potential quality issue for cloned cattle embryos. AB - Nuclear transfer in cattle is associated with high levels of embryonic mortality and often with congenital malformation. Chromosomal abnormalities are a well known cause of pregnancy failure and congenital malformation in humans, but their relative contribution to pregnancy failure and congenital malformation in cloned embryos and calves is largely unknown. This paper reviews existing literature on the chromosomal constitution of bovine embryos produced by fertilization in vivo and in vitro, parthenogenetic activation, and nuclear transfer. The published data suggest that chromosomally abnormal cells are common in embryos; however, the frequency reported varies with the method of embryo production. The most frequently observed deviation from the diploid karyotype was mixoploidy resulting from aberrant cell division causing polyploidy in a variable proportion of the embryo's cells. PMID- 12626099 TI - Fundamental features of chromatin structure. AB - Which mechanisms regulate nuclear plasticity? Part of the answer to that question lies in understanding how genes are expressed and regulated in the context of chromatin structure. It is now clear that the genes are regulated in discrete and controlled stages, from packaging into chromatin to their localization within the nucleus. Whereas the genetic information provides the framework for the manufacture of all proteins necessary to create a living cell, chromatin structure controls how, where, and when the genetic information should be used. In this minireview, I summarize the main characteristics of chromatin structure and highlight some of the modifications usually associated with the regulation of gene expression. PMID- 12626097 TI - SDF-1alpha/CXCR4: a mechanism for hepatic oval cell activation and bone marrow stem cell recruitment to the injured liver of rats. AB - Stromal derived factor-1 alpha (SDF-1alpha) and its receptor CXCR4 have been shown to play a role in the systematic movement of hematopoietic stem cells (HSC) in the fetal and adult stages of hematopoiesis. Under certain physiological conditions liver oval cells can participate in the regeneration of the liver. We have shown that a percentage of oval cells are of hematopoietic origin. Others have shown that bone marrow derived stem cells can participate in liver regeneration as well. In this study we examined the role of SDF-1alpha and its receptor CXCR4 as a possible mechanism for oval cell activation in oval cell aided liver regeneration. In massive liver injury models where oval cell repair is involved hepatocytes up-regulate the expression of SDF-1alpha, a potent chemoattractant for hematopoietic cells. However, when moderate liver injury occurs, proliferation of resident hepatocytes repairs the injury. Under these conditions SDF-1alpha expression is not up-regulated and oval cells are not activated in the liver. In addition, we show that oval cells express CXCR4, the only known receptor for SDF-1alpha. Lastly, in vitro chemotaxis assays demonstrated that oval cells migrate along a SDF-1alpha gradient which suggests that the SDF-1alpha/CXCR4 interaction is a mechanism by which the oval cell compartment could be activated and possibly recruit a second wave of bone marrow stem cells to the injured liver. In conclusion, these experiments begin to shed light on a possible mechanism, which may someday lead to a better understanding of the hepatic and hematopoietic interaction in oval cell aided liver regeneration. PMID- 12626100 TI - Chromatin as a regulative architecture of the early developmental functions of mammalian embryos after fertilization or nuclear transfer. AB - Nuclear transfer of a somatic nucleus into an enucleated oocyte has demonstrated in several mammalian species that the chromatin of a differentiated nucleus can be reprogrammed so as to be able to direct the full development of the reconstructed embryo. This review focus on the timing of the early events that allow the return of somatic chromatin to a totipotent state. Our understanding of the modifications associated with chromatin remodeling is limited by the low amount of biological material available in mammals at early developmental stages and the fact that very few genetic studies have been conducted with nuclear transfer embryos. However, the importance of several factors such as the covalent modifications of DNA through the methylation of CpG dinucleotides, the exchange of histones through a reorganized nuclear membrane, and the interaction between cytoplasmic oocyte components and nuclear complexes in the context of nuclear transfer is becoming clear. A better characterization of the changes in somatic chromatin after nuclear transfer and the identification of oocyte factors or structures that govern the formation of a functional nucleus will help us to understand the relationship between chromatin structure and cellular totipotency. PMID- 12626101 TI - Novel approaches to transdifferentiation. AB - Ways of directly turning a somatic cell into another (a process known as transdifferentiation) would alleviate difficulties associated with current nuclear transplantation procedures and be beneficial for producing replacement cells for therapeutic purposes. Adult stem cells have been shown to display a broader differentiation potential than anticipated and may contribute to tissues other than those in which they reside. In addition, novel transdifferentiation strategies are being developed. We illustrate here a functional reprogramming of a somatic cell using a nuclear and cytoplasmic extract derived from another somatic cell type. Reprogramming of 293T fibroblasts in an extract from T cells is evidenced by nuclear uptake and assembly of transcription factors, induction of activity of a chromatin remodeling complex, changes in chromatin composition and activation of lymphoid cell-specific genes. The reprogrammed cells expressed T cell-specific surface molecules and a complex regulatory function. We propose that in vitro cell reprogramming may create possibilities for producing isogenic replacement cells for therapeutic applications. The system is also likely to constitute a powerful tool to examine the mechanisms of nuclear reprogramming as they occur in vitro. PMID- 12626102 TI - Phenotypic effects of somatic cell cloning in the mouse. AB - Although a variety of phenotypes and epigenetic alterations have been reported in animals cloned from somatic cells, the exact nature and consequences of cloning remain unclear. We cloned mice using fresh or short-term cultures of donor cells (cumulus cells, immature Sertoli cells, and fetal or adult fibroblast cells) with defined genetic backgrounds, and then compared the phenotypic and epigenetic characteristics of the cloned mice with those of fertilization-derived control mice. Irrespective of the nucleus-donor cell type, about 50% of the reconstructed embryos developed to the morula/blastocyst stage, but about 90% of these clones showed arrested development between days 5 and 8, shortly after implantation. Most of the clones were alive at term, readily recovered respiration, and did not show any malformations or overgrowths. However, their placentas were two- to threefold larger than those of the controls, due to hyperplasia of the basal (or spongiotrophoblast) layer. Although there was significant suppression of a subset of both imprinted and non-imprinted placental genes, fetal gene suppression was minimal. The seven imprinted genes that we examined were all expressed correctly from the parental alleles. These findings were consistent for every cell type from the midgestation through term stages. Therefore, cloning by nuclear transfer does not perturb the parent-specific imprinting memory that is established during gametogenesis, and the phenotypic and epigenetic effects of cloning are restricted to placental development at the midgestation and term stages. Twelve male mice that were born in a normal manner following nuclear transfer with immature Sertoli cells (B6D2F1 genetic background) were subjected to long-term observation. They died earlier than the genotype-matched controls (50% survival point: 550 days vs. 1028 days, respectively), most probably due to severe pneumonia, which indicates that unexpected phenotypes can appear as a result of the long-term effects of somatic cell cloning. PMID- 12626103 TI - Effectiveness of off-line and web-based promotion of health information web sites. AB - The relative effectiveness of off-line and web-based promotional activities in increasing the use of health information web sites by target audiences were compared. Visitor sessions were classified according to their method of arrival at the site (referral) as external web site, search engine, or "no referrer" (i.e., visitor arriving at the site by inputting URL or using bookmarks). The number of Australian visitor sessions correlated with no referrer referrals but not web site or search-engine referrals. Results showed that the targeted consumer group is more likely to access the web site as a result of off-line promotional activities. The properties of target audiences likely to influence the effectiveness of off-line versus on-line promotional strategies include the size of the Internet using population of the target audience, their proficiency in the use of the Internet, and the increase in effectiveness of off-line promotional activities when applied to locally defined target audiences. PMID- 12626104 TI - A case study: telemedicine technology and peritoneal dialysis in children. AB - We investigated the feasibility and effectiveness of a telemedicine system for monitoring pediatric patients undergoing automated peritoneal dialysis (APD) at home. The system uses modem-based communication between the patient's cycler and a computer in the dialysis unit, which allows data transmission and storage, and live patient-physician interaction by ISDN lines, modem, microphone with stereo speakers, and digital cameras for private video-conferencing and image capture. Two children aged 10 and 12 years, who live 1500 and 40 km from the dialysis unit, respectively, have been using the system for 7 months. All of the APD treatment data were stored and examined; 122 televisits were performed. The APD data show that both patients have complied with their dialysis prescription. The telemedicine system broadens patient/physician interchange and increases the quality of care and the life of children on peritoneal dialysis. PMID- 12626105 TI - Self-regulation training for chronic pain: can it be done effectively by telemedicine? AB - The purpose of this project was to test the efficacy of behavioral telemedicine interventions designed to teach self-regulation skills to chronic pain patients who had not previously been treated. The study sought to determine whether there were differences in outcome and consumer satisfaction with the pain management intervention when delivered by conventional face-to-face methods, by telephone only, and by closed circuit TV (CCTV). We found that self-regulation techniques can be readily taught via behavioral telemedicine interventions. The primary methodologic weakness of the study is that a single experienced clinician provided all of the interventions. Thus, it is not known at this time how well the intervention can be generalized to other clinicians. In addition, there was no control on the administration of the protocols across conditions. PMID- 12626106 TI - Public information services in the field of communication disorders: comparison between teleservice and E-service. AB - This study evaluates the use of information services in the field of communication disorders by monitoring two communication disorders information services operated in Israel over 8-10 months. One was an E-service and the other was a teleservice. More women than men used the services. Approximately 85% of referrals were judged as appropriate and relevant to the field, and the majority of problems could be treated sufficiently through the service. PMID- 12626107 TI - Toward an understanding of diagnostic teleconsultations and their impact on diagnostic confidence. AB - This paper describes a four-stage model of the medical diagnostic process, and provides the results of an initial test of this emerging knowledge management theory. A study of the diagnostic teleconsultations between primary care physicians located in remote communities and specialists at the Medical College of Georgia was conducted. The study involved an analysis of the video recordings of teleconsultations and postteleconsultation telephone interviews with the specialists and physicians conducting diagnostic teleconsultations. The study revealed that the number of hypotheses generated by specialists, the overall duration of teleconsultations, and physicians' change in diagnostic confidence depended on the diagnostic stage of the specialty teleconsultation. This study demonstrated that the diagnostic stages have different behavioral and cognitive characteristics. Designers of telemedicine programs can complement these characteristics to increase the productivity of health care providers. PMID- 12626108 TI - Proposed model of a digital video-based home telecare system. AB - The purpose of our study was to develop a generic reference model for home telecare in a digital television environment and to suggest how it might be deployed at a test village. We describe here an architecture based on digital video broadcast standards, its components, services, and required additional technical solutions to provide a unified home telecare solution. We also suggest different user groups within the village context and possible service scenarios for them. We conclude that, in general, digital television provides a potentially flexible and promising platform for home telecare services. PMID- 12626111 TI - Lessons from redox signaling in plants. PMID- 12626110 TI - Teleconsultation practice guidelines: report from G8 Global Health Applications Subproject 4. AB - This report presents a series of recommendations derived from deliberations of the G8 countries Subproject 4 Group (SP4 Group) of the Global Health Care Applications Project entitled, A Teleconsultation Practice Guideline. The recommendations provide an initial step toward developing a general guideline platform for the practice of telemedicine/teleconsultation. PMID- 12626109 TI - Health "Smart" home: information technology for patients at home. AB - This article reviews the emerging concept of health "Smart" homes (HSH) and its potential through the use of telemedical information systems and communication technologies. HSH systems provide health care services for people with special needs who wish to remain independent and living in their own home. The large diversity of needs in a home-based patient population requires complex technology. Meeting these needs technically requires the use of a distributed approach and the combination of many hardware and software techniques. We also describe the wide scope of new information, communication, and data-acquisition technologies used in home health care. We offer an introduction to the HSH concept in terms of technical, economic, and human requirements. Examples of HSH projects are presented, including a short description of our own smart home and telehealthcare information system project. PMID- 12626112 TI - Superoxide as an obligatory, catalytic intermediate in photosynthetic reduction of oxygen by adrenaline and dopamine. AB - The superoxide anion radical is known to be the first product of photosynthetic reduction of oxygen mediated by a variety of electron carriers. The effectiveness of many of these electron carriers as herbicides, and the toxicity of the superoxide they produce, have been suggested to rule out oxygen reduction as a physiological component of normal photosynthesis. Here results with isolated spinach chloroplasts are presented that demonstrate that the related catecholamines adrenaline and dopamine mediate photosynthetic reduction of oxygen. Complete inhibition by added superoxide dismutase of light-dependent oxygen uptake by isolated chloroplasts and of the electron transport it supports indicates that superoxide is an obligatory catalytic intermediate, not a product, in adrenaline- and dopamine-mediated oxygen reduction. These compounds might function as chemical analogues of a proposed natural mediator, or oxygen-reducing factor, that allows oxygen reduction to participate in energy transduction in photosynthesis. The identity of the putative natural mediator and the role of oxygen reduction in photosynthesis are discussed. The fully oxidized form of adrenaline, adrenochrome, also acts as a mediator of photosynthetic oxygen uptake, but only by reducing oxygen to superoxide. PMID- 12626113 TI - Characterization of the redox properties of poplar glutaredoxin. AB - The presence of glutaredoxins in plants is now well recognized, but their functions and natural substrates remain largely unknown. Recently, a poplar glutaredoxin has been biochemically characterized and several mutants have been engineered in order to explore its reactivity. This work focuses on some physiological functions of the enzyme. According to our findings, the poplar glutaredoxin can serve as an electron donor to the bacterial 3' phosphoadenylylsulfate reductase as it supports both the catalysis by the enzyme in vitro and complements a methionine auxotroph strain of Escherichia coli. In addition, poplar glutaredoxin is able to reduce the Escherichia coli ribonucleotide reductase 1a (in vitro reduction of cytidine diphosphate). Although this glutaredoxin is described as an electron donor to a phloem-located peroxiredoxin, whose function is to detoxify hydroperoxides, we found that it does not directly reduce hydrogen peroxide or other alkyl hydroperoxides as described for yeast and rice glutaredoxins. However, the poplar glutaredoxin may be involved in the response to oxidative stress as its overexpression in Escherichia coli resulted in a higher resistance toward hydrogen peroxide, menadione, and tert-butyl hydroperoxide. PMID- 12626114 TI - Effects of leaf ascorbate content on defense and photosynthesis gene expression in Arabidopsis thaliana. AB - Ascorbate deficiency in the Arabidopsis thaliana vtc1 mutant had no effect on photosynthesis, but modified defense pathways. The ascorbate content of vtc1 leaves was increased 14-fold after 10 mM ascorbate was supplied, without a concomitant change in redox state. High ascorbate modified the abundance of 495 transcripts. Transcripts encoding dehydroascorbate reductase, pathogenesis related protein 1, and a peroxiredoxin were decreased, whereas those encoding salicylate induction-deficient protein 1, Cu,Zn superoxide dismutase, iron superoxide dismutase, metallothionein, and glutathione transferases were increased. Catalase transcripts were unaffected, but ascorbate peroxidase isoforms APX1 and tAPX were slightly decreased and sAPX transcripts increased. A number of nuclear encoded transcripts for photosynthetic electron transport components were repressed as a result of ascorbate accumulation, whereas those that were chloroplast-encoded were increased. High ascorbate caused decreases in mRNAs encoding chloroplast enzymes such as fructose-1,6-bisphosphatase and sedoheptulose-1,7-bisphosphatase that are activated by reduced thioredoxin. In contrast, others, such as glucose 6-phosphate dehydrogenase, whose activity is inactivated by reduced thioredoxin, were repressed. Together, these results show that ascorbate is involved in metabolic cross-talk between redox-regulated pathways. The abundance of this antioxidant provides information on redox buffering capacity that coordinates redox processes associated with the regulation of photosynthesis and plant defense. PMID- 12626115 TI - The functions of nitric oxide-mediated signaling and changes in gene expression during the hypersensitive response. AB - Nitric oxide (NO) is a highly reactive molecule that rapidly diffuses and permeates cell membranes. In animals, NO is implicated in a number of diverse physiological processes, such as neurotransmission, vascular smooth muscle relaxation, and platelet inhibition. It may have beneficial effects, e.g., as a messenger in immune responses, but it is also potentially toxic when the antioxidant system is overwhelmed and reactive oxygen intermediates (ROI) accumulate. During the last few years, NO has been detected in several plant species, and an increasing number of reports on its function have implicated NO as an important effector in plant growth, development, and defense. The broad chemistry of NO involves an array of interrelated redox forms with different chemical reactivities and numerous potential biological targets in plants. NO signaling functions depend on its reactivity. ROI are key modulators of NO in triggering cell death, but the nature of the mechanisms by which this occurs in plants is different from those commonly observed in animals. This review focuses on the signaling functions of NO, when channeled through the cell death pathway by ROI. PMID- 12626116 TI - Participation of photosynthetic electron transport in production and scavenging of reactive oxygen species. AB - The photosynthetic electron transport chain (PETC) is the principal place of appearance of reactive oxygen species (ROS) in plants under illumination. The peculiarities of this process in different segments of the PETC are discussed. Oxygen uptake observed under impaired electron donation to photosystem II is attributed mainly to hydroperoxide formation by reaction of oxygen with organic radicals generated after detachment of electrons by P680(+). Oxygen reduction in the plastoquinone pool is suggested to start with the reaction of O(2) with plastosemiquinone, and to be followed by reduction of superoxide to hydrogen peroxide by plastohydroquinone. The distribution of plastoquinone throughout the thylakoid membrane interior provides for the generation of ROS by this route all along the membrane surface. O(2) reduction at the acceptor side of photosystem I remains poorly understood. The regeneration of antioxidants is stated to be a priority task of photosynthetic electron transport in view of the effectiveness of monodehydroascorbate as electron acceptor. We propose that ROS generation in the plastoquinone pool and the possible formation of hydroperoxides in the vicinity of photosystem II are key processes participating in the primary stages of redox signaling. PMID- 12626117 TI - Redox regulation of thylakoid protein phosphorylation. AB - The photosystem II of chloroplast thylakoid membranes contains several proteins phosphorylated by redox-activated protein kinases. The mechanism of the reversible activation of the light-harvesting antenna complex II (LHCII) kinase(s) is one of the best understood and related to the regulation of energy transfer to photosystem II or I, thereby optimizing their relative excitation (state transition). The deactivated LHCII protein kinase(s) is associated with cytochrome b(6)f and dissociates from the complex upon activation. Activation of the LHCII protein kinase occurs via dynamic conformational changes in the cytochrome b(6)f complex taking place during plastoquinol oxidation. Deactivation of the kinase involves its reassociation with an oxidized cytochrome complex. A fine-tuning redox-dependent regulatory loop inhibits the activation of the kinase via reduction of protein disulfide groups, possibly involving the thioredoxin complex. Phosphorylation of LHCII is further modulated by light-induced conformational changes of the LHCII substrate. The reversible phosphorylation of LHCII and other thylakoid phosphoproteins, catalyzed by respective kinases and phosphatases, is under strict regulation in response to environmental changes. PMID- 12626118 TI - Redox signaling in the chloroplast: the ferredoxin/thioredoxin system. AB - Chloroplasts have developed a light-dependent system for the control of the activities of key enzymes involved in assimilatory (photosynthetic) and dissimilatory pathways, which allows a switch between these opposing pathways to prevent futile cycling. This regulatory system, known as the ferredoxin/thioredoxin system, consists of several proteins constituting a redox cascade that transmits the light signal perceived by chlorophyll to selected target proteins, thereby influencing their activity. A central component of the redox cascade is a novel enzyme, the ferredoxin:thioredoxin reductase, which is capable of reducing a disulfide bridge with the help of an iron-sulfur cluster. Recent developments on the elucidation of the structures of several implicated proteins and on the mechanism of signal transfer have greatly improved our understanding of this regulatory mechanism. This review describes the components of the redox cascade, the principal target proteins, and the mechanism of action of the light-signal transfer. PMID- 12626119 TI - Redox regulation of chloroplast transcription. AB - Chloroplasts are the important plant cell organelles where photosynthesis takes place. Throughout this process, reaction center proteins are degraded and subsequently replenished by redox-responsive gene expression. In addition to well defined posttranscriptional mechanisms at the RNA and protein level, the transcription of chloroplast DNA into RNA precursors has been a focal point of studies in this area. Evidence has become available for a central role of a redox responsive protein kinase named plastid transcription kinase (PTK) because of its association with the chloroplast transcription complex. The recent cloning of the PTK gene has resulted in a full-length cDNA for a protein related to the catalytic alpha subunit of nucleocytoplasmic casein kinase (CK2), yet with an additional chloroplast transit peptide. The corresponding protein, termed cpCK2alpha, was shown to be associated with the major organellar RNA polymerase, PEP-A. Both authentic PTK and recombinant cpCK2alpha have comparable general properties in vitro, and both are subject to regulation by the redox-reactive reagent glutathione. Based on the physical and functional equivalence, it is anticipated that the cloned protein can help clarify the functional role of the transcription kinase in vivo, including the identification of interaction partners at the interface between photosynthetic redox signaling and gene expression. PMID- 12626120 TI - Redox control of posttranscriptional processes in the chloroplast. AB - The ability to couple photosynthetic electron transport and redox poise to plastid gene expression enables plants to respond to environmental conditions and coordinate nuclear and chloroplast activities in order to maintain photosynthetic efficiency. The plastid redox regulatory system serves as a paradigm for understanding redox-regulated gene expression. In this review, we will focus on posttranscriptional events of redox-regulated gene expression in the chloroplast. As redox regulation of enzymatic activities in the chloroplast will be covered in other reviews in this volume, as will discussions on the redox regulation of chloroplast transcription, we will concentrate on the available evidence for redox regulation of chloroplast translation, and mRNA splicing and turnover. PMID- 12626121 TI - Chloroplast redox control of nuclear gene expression--a new class of plastid signals in interorganellar communication. AB - Chloroplasts are genetically semiautonomous organelles that contain their own subset of 100-120 genes coding for chloroplast proteins, tRNAs, and rRNAs. However, the great majority of the chloroplast proteins are encoded in the nucleus and must be imported into the organelle after their translation in the cytosol. This arrangement requires a high degree of coordination between the gene expression machineries in chloroplasts and nucleus, which is achieved by a permanent exchange of information between both compartments. The existence of such coordinating signals has long been known; however, the underlying molecular mechanisms and signaling routes are not understood. The present data indicate that the expression of nuclear-encoded chloroplast proteins is coupled to the functional state of the chloroplasts. Photosynthesis, which is the major function of chloroplasts, plays a crucial role in this context. Changes in the reduction/oxidation (redox) state of components of the photosynthetic machinery act as signals, which regulate the expression of chloroplast proteins in both chloroplasts and nucleus and help to coordinate the expression both in compartments. Recent advances in understanding chloroplast redox regulation of nuclear gene expression are summarized, and the importance for intracellular signaling is discussed. PMID- 12626122 TI - Reactive quinones differentially regulate SAPK/JNK and p38/mHOG stress kinases. AB - The stress-activated protein kinases SAPK/JNK and p38/mHOG are activated by diverse classes of stress stimuli, many of which induce redox perturbations. We studied the effects of reactive quinones on stress signaling pathways. Menadione (2-methyl-1,4-naphthoquinone), which undergoes both one- and two-electron reduction, completely inhibited SAPK activity at high concentrations while activating SAPK at lower concentrations. Menadione activated p38/mHOG dose responsively. 2,3-Dimethyl-1,4-naphthoquinone (DMNQ), which preferentially undergoes two-electron reduction, had similar effects. In contrast, 1,4 naphthoquinone, which preferentially undergoes one-electron reduction, inhibited SAPK at high concentrations, but failed to activate SAPK at any concentration tested. In addition, this quinone activated p38 only at lower concentrations; high concentrations inhibited p38 activity. These activity profiles correlated with the activation state of the upstream kinase, indicating that the effects were mediated by an upstream step in the kinase pathway. The quinone reductase inhibitor dicoumarol blocked activation of SAPK by menadione and DMNQ, suggesting that two-electron reduction is important. Finally, addition of increasing amounts of hydrogen peroxide mimicked the effects of menadione and DMNQ, suggesting that hydrogen peroxide may be the relevant mediator. Differential activation of stress kinases by reactive quinones demonstrates that the cellular redox environment independently modulates these pathways. PMID- 12626123 TI - Involvement of reactive oxygen species in cardiac preconditioning in rats. AB - To date, the involvement of reactive oxygen species in ischemic preconditioning in vivo in rats is not clearly demonstrated. The aim of the present study was to determine whether N-(2-mercaptopropionyl)glycine (MPG), a cell-diffusible hydroxyl radical scavenger, and carnosine, a potent singlet oxygen quencher, could block protection afforded by a single cycle of ischemic preconditioning in vivo in the rat. An ESR study was first performed to validate in vitro the specific antioxidant properties of carnosine and MPG. In a second set of experiments, open-chest rats were subjected to 30 min of left coronary occlusion followed by 60 min of reperfusion. Preconditioning was elicited by 5 min of ischemia and 5 min of reperfusion. Neither MPG (1-h infusion, 20 mg/kg) nor carnosine injection (bolus, 25 micro mol/rat) affected infarct size. The infarct size-limiting effect of preconditioning was completely blunted by MPG, whereas carnosine did not alter the cardioprotection. It is concluded that free radicals and especially hydroxyl radicals could be involved in the adaptive mechanisms induced by a single cycle of preconditioning in vivo in rats. PMID- 12626124 TI - Regulation of vascular endothelial growth factor synthesis by nitric oxide: facts and controversies. AB - Vascular endothelial growth factor (VEGF) is the major molecule governing angiogenesis, defined as the growth of blood vessels from vascular structure. There is abundant evidence that nitric oxide (NO) is an effector molecule mediating the activity of VEGF. By binding to its receptors, VEGF initiates the signaling cascades leading to NO production and angiogenic activation of endothelial cells. Recent data show that NO induces VEGF synthesis in numerous cell types, including vascular smooth muscle cells, macrophages, keratinocytes, and tumor cells. NO enhances VEGF production by augmenting its expression through activation of Akt kinase, followed by induction of several transcription factors, of which stabilization of hypoxia-inducible factor (HIF-1) is the critical step. With respect to its effect on VEGF expression, NO mimics hypoxia, the classical activator of HIF-1 and VEGF synthesis. The effect of NO on VEGF production is also mediated by heme oxygenase, an enzyme generating carbon monoxide, which appears to stimulate VEGF release. In this review, we attempt to elucidate the molecular mechanisms underlying the effects of NO on VEGF synthesis. We also discuss some discrepant data and suggest explanations for various aspects of the NO-VEGF relationship. PMID- 12626125 TI - Redox control on the cell surface: implications for HIV-1 entry. AB - Proteins that work outside cells nearly always contain disulfide bonds. The prevailing view is that these bonds have been added during evolution to enhance protein stability. Recent evidence suggests that disulfide bonds can also control protein function. Certain secreted proteins contain one or more disulfide bonds that can control function by breaking and reforming in a controlled way. This review focuses on disulfide exchange events on the cell surface, with a particular reference to two proteins involved in HIV-1 infection. The primary HIV 1 receptor on immune cells, CD4, and the viral envelope glycoprotein, gp120, play a central role in HIV-1 entry. Redox change in a disulfide bond or bonds in one or both of these proteins appears to be important for HIV-1 entry. PMID- 12626126 TI - Astaxanthin limits exercise-induced skeletal and cardiac muscle damage in mice. AB - Dietary antioxidants may attenuate oxidative damage from strenuous exercise in various tissues. Beneficial effects of the antioxidant astaxanthin have been demonstrated in vitro, but not yet in vivo. We investigated the effect of dietary supplementation with astaxanthin on oxidative damage induced by strenuous exercise in mouse gastrocnemius and heart. C57BL/6 mice (7 weeks old) were divided into groups: rested control, intense exercise, and exercise with astaxanthin supplementation. After 3 weeks of exercise acclimation, both exercise groups ran on a treadmill at 28 m/min until exhaustion. Exercise-increased 4 hydroxy-2-nonenal-modified protein and 8-hydroxy-2'-deoxyguanosine in gastrocnemius and heart were blunted in the astaxanthin group. Increases in plasma creatine kinase activity, and in myeloperoxidase activity in gastrocnemius and heart, also were lessened by astaxanthin. Astaxanthin showed accumulation in gastrocnemius and heart from the 3 week supplementation. Astaxanthin can attenuate exercise-induced damage in mouse skeletal muscle and heart, including an associated neutrophil infiltration that induces further damage. PMID- 12626127 TI - Anti-vascular endothelial growth factor antibody and nimustine as combined therapy: effects on tumour growth and angiogenesis in human glioblastoma xenografts. AB - We evaluated the effectiveness of vascular endothelial growth factor (VEGF) blockade alone and in combination with 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3 (2-chloroethyl)-3-nitrosourea (ACNU, nimustine), a cytotoxic agent commonly used in the treatment of malignant gliomas, to eradicate tumors of human glioblastoma cell lines implanted in SCID (severe combined immunodeficiency) mice. ACNU, but not cisplatin and etoposide, elevated VEGF expression in a glioma cell line in vitro. VEGF antibody alone inhibited glioma growth in vivo as a result of angiogenesis inhibition. The combination with ACNU resulted in an additive effect for inhibition of glioma growth. ACNU also induced VEGF up-regulation in glioma tissues, which was decreased with VEGF antibody treatment. One of the mechanisms of the additive effect of the VEGF antibody and ACNU combination is the blockade of VEGF up-regulation induced by ACNU. As such, the combination of antiangiogenic therapy with conventional therapy is promising for glioma treatment in the future. PMID- 12626128 TI - A phase I study of topotecan as a radiosensitizer for brainstem glioma of childhood: first report of the Children's Cancer Group-0952. AB - Our purpose was to establish the maximum tolerated dosage (MTD) of daily i.v. topotecan with conventionally fractionated radiotherapy (XRT) for patients with intrinsic pontine glioma of childhood. Topotecan was given as a 30-min i.v. infusion 30-60 min before each XRT treatment given daily for 33 days. Total XRT dose was 59.4 Gy. Dose escalation of topotecan was carried out using a standard phase I design. Dose limiting toxicity (DLT) was defined as an absolute neutrophil count (ANC) of < or =500/mm(3) for > or =7 days; platelets of < or =50,000/mm(3) for > or =7 days; >7 days platelet transfusions; fever and neutropenia (ANC < or =500/mm(3) for > or =7 days); and/or any > or=grade 3 non hematologic toxicity. In this multi-institutional phase I study, 17 patients <21 years with intrinsic pontine glioma were enrolled. Sixteen patients completed treatment. An ANC < or =500/mm(3) for > or =7 days occurred in 2/5 patients at 0.50 mg/m(2) of topotecan, which was the DLT. The remaining 14 patients received topotecan without experiencing DLT. One patient at 0.40 mg/m(2) died of disease progression while on treatment. There were 6 other grade 4 hematologic events (5 ANCs <500/mm(3), 1 hemoglobin <6. 5 g/dl) not meeting DLT criteria. No significant non-hematologic toxicities were seen. The actuarial median survival time is 15 months (95% confidence interval, 9.6-19 months); 1-year survival is 53%. DLT of daily topotecan with cranial XRT is grade 4 neutropenia for > or =7 days at 0.50 mg/m(2) x 33 (total dosage = 16.5 mg/m(2)); the recommended safe MTD of daily topotecan for further phase II testing is 0.40 mg/m(2) x 33 (total dosage = 13.2 mg/m(2)). PMID- 12626130 TI - Are all psychological treatments for psychosis equal? The need for CBT in the treatment of psychosis and not for psychodynamic psychotherapy. AB - Paley and Shapiro conclude in their review (Paley & Shapiro, 2002) that there is clear evidence that cognitive-behaviour therapy (CBT) results in clear clinical benefits for people with schizophrenia. They argue that this research could be informed by the wider psychotherapy literature, specifically the 'equivalence paradox' and 'investigator allegiance', and that psychodynamic treatments for schizophrenia should be investigated and evaluated. We argue that their review of the literature is incomplete. We conclude that there is little evidence to support the 'equivalence paradox' generally, and where it is suggested, as in depression, this is most likely an artefact. In schizophrenia, there is consistent evidence that supportive psychotherapy performs poorly in treating hallucinations. We argue for improved methodology in carrying out randomized controlled trials, which would minimize bias, such as from 'investigator allegiance'. We consider evaluation of psychodynamic psychotherapy to demonstrate equivalence to CBT as a low priority and unlikely ever to be attempted. We consider the research and clinical priorities to be the further developments of CBT and family intervention methods, which could have significant benefits for sufferers of schizophrenia and their carers. PMID- 12626129 TI - Biologic characterization of a secondary glioblastoma with extracranial progression and systemic metastasis. AB - Glioblastomas rarely metastasize outside the CNS. We biologically characterized a case of secondary glioblastoma associated with extracranial progression and distant metastasis. A 42-year-old male patient was subjected to craniotomy for a left temporal tumor (astrocytoma grade II) and subsequently underwent another 3 craniotomies due to tumor recurrences. At the third craniotomy, extracranial progression was noted, and the tumor was classified as a glioblastoma. In order to pinpoint the genes expressed differentially in the intracranial primary tumor and the metastatic tumors, we used cDNA microarray. The patterns of gene expression in these 2 samples were highly similar, suggesting that the mechanism of metastasis was direct infiltration of tumor cells into extracranial blood vessels. Insulin-like growth factor binding protein-2 was overexpressed in both primary and metastatic tumors. Immunohistochemical studies of DNA-dependent protein kinase, which participates in the repair of DNA, was strongly positive in the samples obtained at the first and second operations, but the positive rates were markedly reduced in the specimens obtained at the third and fourth operations. These results suggest that insulin-like growth factor binding protein 2 and deficiency of DNA-dependent protein kinase proteins promoted tumor progression in the present case. PMID- 12626132 TI - A cognitive therapy model of social anxiety problems: potential limits on its effectiveness? AB - Cognitive therapy formulations of, and interventions for, social phobia are increasingly influential. In this article, relevant studies are reviewed and lead to a tentative conclusion that purely 'cognitive' interventions may, in fact, fail to provide optimum help for a large number of people likely to present to mental health services with problems of social anxiety. PMID- 12626131 TI - Patients' relationship episodes and therapists' feelings. AB - The idea that psychotherapists' feelings may reflect some aspect of the content in the patients' material has long been clinically accepted but on the whole poorly systematically studied. The aim of this study was to analyse associations between relationship episodes told by patients at evaluation interviews, and therapists' subsequent feelings towards the patients. In total, 28 psychotherapies with nine psychotherapists were studied. Before therapy started, the clients were interviewed separately by the eventual therapist and a second interviewer and six relationship episodes were written down by the interviewers. During the therapy, the therapists reported their feelings on a 'feeling checklist' after each therapy session. In a first analysis, the relationship episodes were categorized according to the CCRT system. The results indicated only scattered associations between CCRT categories and therapist feelings. The results were, however, difficult to interpret as it was found that the therapists' feeling patterns were quite homogenous for each therapist. Considering this, qualitative analyses were made of six therapies carried out by two therapists. These analyses revealed different but theoretically plausible patterns between the feelings attributed to the persons in the patients' relationship episodes and the therapists' subsequent feelings towards the patients. The results were interpreted as a confirmation of the purported relations between patient relationship episodes and therapist feelings, but also as pointing to the need to develop the understanding of these patterns. It seems particularly important to emphasize the importance of the therapist's own habitual feeling patterns when the impact of the patient's internalized relationship patterns on countertransference reactions is evaluated. PMID- 12626133 TI - Study of psychiatric admission patterns before and after out-patient psychotherapy in a district service setting. AB - There is a need for evidence to demonstrate the clinical and economic effectiveness of psychotherapy as a component of mental health service delivery. A naturalistic cohort study was carried out in a specialist psychotherapy department in a district mental health service. Thirty-six patients were identified with hospital admissions in the study period who had also received a course of outpatient psychotherapy. There was a significant decrease in hospital in-patient usage temporally associated with receipt of psychotherapy and associated cost savings. Out-patient psychotherapy leads to less in-patient bed usage and significant health-care savings among previously hospitalized psychiatric patients (approximately 6700 pounds sterling per patient). PMID- 12626134 TI - Pathways to anaclitic and introjective depression. AB - Depression may be conceptualized multidimensionally, owing to marked individual differences in developmental vulnerability and phenomenology. Blatt (Blatt, Quinlan, Chevran, McDonald, Zuroff, 1982; Blatt, Quinlan, Pilkonis, & Shea, 1995) has validated two subtypes of depression. Anaclitic depression involves excessive interpersonal concerns, including feelings of loneliness, weakness, helplessness and abandonment fears. Introjective depression denotes achievement concerns, and is characterized by a tendency towards self-criticism and self-evaluation. Present research examined this distinction by identifying possible differential attachment patterns for the depression subtypes. It was hypothesized that, while secure attachment would negatively relate to depression, insecure attachment would predict anaclitic and introjective subtypes, with perfectionism mediating this relationship. A sample of 245 participants completed the Depressive Experiences Questionnaire, the Beck Depression Inventory, the Multidimensional Perfectionism Questionnaire, and two measures of attachment. Regressional techniques revealed that anaclitic depression was predicted by preoccupied attachment (seeking others' acceptance to bolster low self-worth), with socially prescribed perfectionism (striving to meet the high standards of others) partially mediating the relationship. Introjective depression was predicted by fearful-avoidant attachment (low self-worth and expectations of rejection), with self-oriented perfectionism (self-imposed unrealistic standards) acting as a partial mediator. Findings further validate these depressive subtypes by indicating their different attachment-perfectionism paths. Results are concordant with attachment theory and have implications for both theory and therapy. PMID- 12626135 TI - Investigating individuals' motivations to become counselling psychologists: The influence of early caretaking roles within the family. AB - This study examines the early family experiences of a sample of trainee counselling psychologists in order to gain insight into their career motivations. Specifically, it was hypothesized that counselling students would be more likely to report early occupation of caring roles within the family and less experience of appropriate bonding with their parents compared with students training to enter a non-counselling psychology profession. Participants completed scales to assess Parentification, Parental Bonding and Self-efficacy toward Caring. The responses of 64 students were analysed, 32 in each respective group. Results indicated that counselling psychology trainees reported significantly lower levels of care from their mothers but reported higher levels of self-efficacy toward caring. An investigation of the predictors of parentification indicated that a lack of care from fathers was associated with higher parentification scores for all participants. Care from mothers was shown to relate differently to parentification, depending on the gender of the participant, such that a lack of care from mothers was related to parentification for female participants, whereas greater care from mothers was related to parentification for male participants. Implications for counselling psychology training and practice are discussed in addition to future directions for parentification research. PMID- 12626137 TI - The legal situation regarding assisted reproduction in Germany. AB - Current and proposed laws regarding the practice of assisted conception and preimplantation genetic diagnosis (PGD) in Germany are discussed in this paper. Strict limits on scientific and clinical studies into assisted conception that were imposed by legislators are analysed in response to modern trends. New laws are in prospect, and discussions on their form are attracting much debate. Ethical issues cover donor insemination and surrogate motherhood, and increasingly involve intracytoplasmic sperm injection. The limit of three fertilized oocytes, and cryopreservation of pronucleate eggs only, will probably be retained in any new law. Proposals for PGD are still attracting considerable opposition, in relation to the concept of totipotency that underlined earlier legislation. These attitudes are examined and criticized in this review, and comments that such techniques infringe personal dignity are questioned in some detail. So too are proposals to limit clinicians' liability for a child being born following failure of sterilization, with a recommendation to avoid performing this surgery on the grounds of avoiding future financial burdens for the couple. Possible future legislation is considered briefly at the conclusion of the article. PMID- 12626139 TI - Roles of cyclic GMP in modulating ovarian functions. AB - The production of a viable oocyte is dependent upon the critical influences of gonadotrophins on follicular development, granulosa cell maturation, ovulation, and luteinization. While the effects of LH and FSH are due in large part to cyclic AMP-dependent signalling mechanisms, it is clear that a number of other factors modulate the actions of gonadotrophins on the ovary via activation of alternative signalling pathways. In this regard, recent studies indicate that the second messenger guanosine 3',5'-cyclic monophosphate (cGMP) mediates a wide range of influences on the ovary. Nitric oxide (NO) is a major regulator of cGMP production via its action on soluble guanylyl cyclase, while natriuretic peptides activate receptors with intrinsic guanylyl cyclase activities. In addition, other factors known to influence ovarian functions are now recognized to act via NO/cGMP pathways. This report will review these previous findings and present new data demonstrating the inhibitory influence of cGMP on cAMP-stimulated LH receptor expression in cultured granulosa cells. PMID- 12626140 TI - Apoptosis in the human ovary. AB - Programmed cell death or apoptosis is an essential component of human ovarian function and development. During early fetal life approximately 7 x 10(6) oocytes are formed in the human ovary. However, the number of oocytes is dramatically reduced already before birth through apoptotic cell death. In reproductive life, a number of primordial follicles start growing during each menstrual cycle. Usually only one will ovulate and the fate of the rest of the follicles is atresia through the mechanism of apoptosis. Ultimately, only around 400 follicles will ovulate during a woman's reproductive life. After ovulation, the dominant follicle forms the corpus luteum, a novel endocrine gland that is responsible for the production of progesterone and maintenance of endometrium during early pregnancy. If pregnancy does not occur, corpus luteum action must cease so that new follicles can resume growing during the next menstrual cycle. Apoptosis is also responsible for corpus luteum regression in the human ovary. In recent years, new knowledge of the role and regulation of apoptosis in the ovary has been acquired through the use of knockout and transgenic animals. Apoptosis seems to be the mechanism that makes the female biological clock tick. The following review will discuss the role of apoptosis and its regulation of human ovarian function. PMID- 12626141 TI - The polycystic ovary syndrome: what does insulin resistance have to do with it? AB - About half of infertility cases are attributable to female factors, of which anovulation is the leading cause. Most cases of anovulation are due to the polycystic ovarian syndrome (PCOS). Clinically PCOS, present in 5% of all women, consists of anovulation, acne, hirsutism, reversed LH-to-FSH ratio and often resistance to insulin. In some cases PCOS appears in familial clusters but its genetic cause is unknown. Several genes were suggested as possible culprits for PCOS but their involvement has not been proven. The central paradox regarding the role of insulin in PCOS is the high responsiveness to insulin by the ovary, as opposed to the resistance of the whole body. On the backdrop of knowledge of several paralogous genes for each of the participating proteins in the insulin signal transduction pathways, it is possible that different paralogues propagate the intracellular signal in the ovary as opposed to peripheral tissues. Studies by microarray techniques of the different gene expression profiles in the two ovarian cells and peripheral cells such as adipocytes could clarify whether the ovarian defect in PCOS is identical to the peripheral defect in insulin signal transduction, or whether serum insulin concentrations simply serve to reveal the ovarian defect. PMID- 12626142 TI - Gonadotrophin-releasing hormone antagonists: will they replace the agonists? AB - Modern gonadotrophin-releasing hormone (GnRH) antagonists such as cetrorelix and ganirelix reliably prevent premature LH surges in controlled ovarian hyperstimulation for assisted reproductive technologies. Cetrorelix and ganirelix are safe and effective compounds. Because of their distinct pharmacological mode of action, it has been possible to achieve a significant reduction of treatment time. Fertilization and pregnancy rates are comparable to those obtained in agonist protocols for ovarian stimulation. No allergic or hyperergic reactions have been reported, and patient compliance is excellent. The fact that GnRH antagonists allow an immediate suppression of gonadotrophin concentrations while preserving pituitary responsiveness to endogenous GnRH provides enormous flexibility in treatment. GnRH antagonists have helped to overcome some major disadvantages of GnRH agonists, especially of the long protocol, which is currently the standard protocol for ovarian stimulation. The mistaken administration of antagonists during early pregnancy is not possible, since they are only administered during ovarian stimulation when a premature LH surge may be imminent. They are used in the spontaneous cycle or after pretreatment with oral contraceptives. Pregnancy can easily be ruled out by testing for human gonadotrophic hormone before onset of gonadotrophin stimulation on the second or third day of the cycle. Since flare-up effects are absent, there is no risk of cyst formation. Hormonal withdrawal symptoms are eliminated, since no period of pituitary suppression occurs, and therefore exogenous gonadotrophins are not required. Overall, the duration of the stimulation cycle is as short as a normal menstrual cycle. The procedure seems to be safer than the long protocol, since the most serious complication, the occurrence of severe cases of OHSS, is reduced. It is also safe with respect to the course of pregnancies and the health of offspring. Both protocols developed so far, the single dose and multiple dose antagonist protocol, are comparable, utilizing data from the large prospective phase IIIb studies. Although several studies have indicated a slight reduction in pregnancy rate with GnRH antagonists as compared with agonists, this problem may be rectified by developing flexible antagonist regimens designed for individual patients. Introducing flexible GnRH antagonist regimens should be the area of research in the near future. PMID- 12626143 TI - Chromosomal abnormalities in a series of 6,733 human oocytes in preimplantation diagnosis for age-related aneuploidies. AB - Most chromosomal abnormalities originate from female meiosis and contribute significantly to pregnancy failures, particularly in women of advanced maternal age. A total of 8,382 oocytes were obtained in 1,297 IVF cycles from patients of advanced maternal age (mean 38.5 years). Following a standard IVF protocol, oocytes were tested following removal and fluorescence in-situ hybridization (FISH) analysis of the first (PB1) and second polar bodies (PB2), using probes specific for chromosomes 13, 16, 18, 21 and 22 (Vysis). FISH results were available in 67,33 (80.3%) oocytes tested, 3,509 (52.1%) of which were aneuploid, with the remaining 3,224 (47.9%) normal oocytes available for transfer. In all, 41.7% of oocytes had meiosis I errors, compared to 35.1% with meiosis II errors. Abnormalities in meiosis I were represented by extra chromatids in 15.4%, missing chromatids in 48.1%, missing chromosomes in 5.9%, extra chromosomes in 0.5%, and complex abnormalities in 30.1%. The proportions of abnormal oocytes with missing or extra chromatids in meiosis II were 36.6 and 41.2% respectively, with the remaining oocytes having complex abnormalities, involving missing or extra chromatids of different chromosomes (22.1%) following meiosis II. Overall, 41.8% oocytes had meiosis I, 30.7% meiosis II, and 27.6% both meiotic division errors. A total of 45.1% of the abnormal oocytes had complex errors, involving the same chromosome in both meiotic divisions (21.5%), or different chromosomes (78.5%), of which 74.8% were with abnormalities of two, and 25.2% with abnormalities of three chromosomes studied. Of 3,224 detected aneuploidy-free zygotes, 2,587 were transferred in 1,100 treatment cycles (2.35 embryos per transfer), resulting in 241 (21.9%) clinical pregnancies and 176 healthy children born, suggesting a positive clinical outcome following aneuploidy testing of oocytes in a group of IVF patients of average age 38.5 years. PMID- 12626144 TI - Somatic cell haploidization: an update. AB - Oocyte donation is the only method of treating female sterility caused by complete absence of oocytes, with the loss of genetic motherhood. Genetic fatherhood of males with complete absence of spermatozoa can only be restored by assisted reproduction treatment if sperm precursor cells belonging to the male germline can still be recovered from the testis. Otherwise, sperm donation is the only available solution. Somatic nucleus haploidization after injection into previously enucleated donor oocytes (diploid-to-haploid reduction) might enable the reconstruction of new oocytes carrying the complete nuclear genome of female patients lacking their own oocytes. Such newly formed oocytes could subsequently be fertilized by spermatozoa from the patient's husband. In cases of male infertility with complete absence of the germline, the patient's somatic cell nuclei could be injected into the oocytes without previous enucleation, and somatic nucleus haploidization would occur in the presence of the original female nucleus (triploid-to-diploid reduction), hopefully leading to the formation of a diploid embryo. Both interventions differ substantially from cloning because embryos are formed by syngamy with the male and female genomes originating from the two genetic parents, as in natural fertilization. Ultrastructural remodelling of mouse somatic cell nucleoli can be achieved in enucleated metaphase II mouse oocytes. Haploidization has also been attempted with Sertoli cells and with fibroblasts, both of which are also available in male patients. Experiments are currently under way to assess the regularity of chromatid segregation during somatic nucleus haploidization. PMID- 12626145 TI - Birth resulting from transfer of blastocysts cryopreserved with propanediol after spontaneous hatching. AB - This paper reports the birth of a healthy baby resulting from transfer of blastocysts that were cryopreserved using propanediol after spontaneous hatching. A young infertile couple underwent IVF treatment in the clinic. After several IVF attempts, two births resulted; the first one with fresh embryos in 1996 after three IVF cycles, and the second one in 1999 (after a new IVF cycle in 1998) with frozen blastocysts that had remained cryopreserved in 1.5 mol/l propanediol and 0.1 mol/l sucrose after spontaneous hatching. This report of a healthy baby following transfer of hatched blastocysts frozen in propanediol supports further exploration of this approach. PMID- 12626146 TI - Counselling on cryopreservation of pronucleated oocytes. AB - The German Embryo Protection Law permits only the cryopreservation of supernumerary pronuclear stage eggs. Data are needed to counsel patients about their individual benefits from this procedure. All fresh embryo transfer cycles performed from January 1994 until December 1998 in which supernumerary pronucleate eggs were cryopreserved (n = 557) were analysed retrospectively, together with data from all subsequent cycles involving transfers of frozen thawed pronucleate eggs (n = 420) from January 1994 until June 2001. The additional cumulative pregnancy rate per fresh cycle was 11.5%. This rate depended on the number of embryos per transfer, i.e. 1.9, 8.2 and 13.0% respectively when one, two or three embryos were transferred (P < 0.05). A strong correlation was found between the numbers of cryopreserved pronucleate eggs and pregnancy rates, of 9.3, 10.5 and 17.1% when 1-3, 4-6, or at least 7 pronucleate eggs were available respectively. Additional benefit in terms of this rate from cryopreservation for a patient with and without a pregnancy in the fresh embryo transfer cycles was 5.3 and 12.7% respectively. It is concluded that higher pregnancy rates in cycles involving cryopreserved eggs for patients who did not become pregnant in fresh transfer cycles reveals the disadvantage of the German Embryo Protection Law, which does not allow embryo selection. Therefore, cryopreservation of pronucleate eggs with a higher developmental potential is possible. The total pregnancy rate could be raised from 28.0 to 35.5% per fresh transfer cycle. PMID- 12626147 TI - Counselling lesbian couples: requests for donor insemination on social grounds. AB - Fertility centres are increasingly involved in dealing with requests from lesbian couples for donor insemination (DI). Data were collected on 95 Belgian lesbian couples who applied for DI. The majority of couples were well integrated in a social environment (family, friends and work) that consisted mainly of heterosexuals. They tended to be open about their homosexuality and most couples considered the social environment to be tolerant towards their homosexual orientation. Couples who considered alternatives to DI would have liked to have had more information on the donor and were more inclined to introduce a 'godfather' who would take special interest in the child. Couples who considered DI to be the best solution considered the absence of a father to be less of a problem for the child and wished to have no information at all as regards the donor. Approximately half the couples considered their family a two-mother unit. For the other couples, the family unit consisted of a mother and her partner who shared parental responsibility for the child equally. The issues that are considered important to cope with and on which lesbian couples should be counselled are presented in the discussion. PMID- 12626148 TI - Oxygen, early embryonic metabolism and free radical-mediated embryopathies. AB - Free radicals, once the preserve of chemists, are now recognized as playing a central role in many biological systems. They are formed as an inevitable by product of aerobic respiration and various cytoplasmic processes at a rate dependent upon the prevailing oxygen tension. At physiological concentrations, oxygen and nitrogen free radical species play key roles in intracellular signalling, regulating many homeostatic mechanisms and mediating stress responses. If concentrations exceed cellular defences, however, then indiscriminate damage may occur to lipids, proteins and DNA. Cell function may be perturbed, and in the most severe cases apoptosis may result. Although there are significant species differences, many aspects of early mammalian development, from fertilization through to differentiation of the principal organ systems, take place in vivo in a low oxygen environment. This may serve to protect the embryo from free radical damage, for exposure of early embryos to ambient oxygen concentrations or the products of maternal metabolic disorders is often associated with reduced viability and an increased rate of congenital malformations. Administration of free radical scavengers, including vitamins C and E, can mitigate many of these effects, indicating the importance of a balanced maternal diet to successful reproduction. PMID- 12626149 TI - Aspects of the molecular regulation of early mammalian development. AB - Many regulatory systems operate in the early mammalian embryo. This brief overview surveys several systems and their integration including polarities and axes, left-right differentiation, timers in cells, tissues and in gene expression, and imprinting. Polarities are essential from the very earliest stages of oocyte formation, and maintain their significance until blastocyst stages and beyond. They determine cleavage axes and the distribution of maternal proteins in the oocyte, distinct distributions being identified at the animal pole especially. Left-right axes are no doubt expressed from the earliest embryonic stages, and perhaps even in determining slight differences in the axes of cleavage and of maternal protein distribution. Timers, equally fundamental, have been demonstrated to control many functions in oocytes and embryos. Many fundamental processes in early mammalian oocytes and embryos are closely timed. They are classified into circadian rhythms, hourglass timers, clocks regulating major aspects of development including transcription, longevity via telomere clocks and long-range systems. Imprinting and methylation, increasingly important in establishing stable phenotypes in early embryos, might develop abnormally under some circumstances including intracytoplasmic sperm injection and cloning. A general summary briefly describes some other aspects of regulation, especially chromosomal anomalies in human embryos. PMID- 12626150 TI - Proposed Canadian legislation to regulate reproductive technologies and related research. PMID- 12626168 TI - Progestogen-only emergency contraception and ectopic pregnancy. PMID- 12626169 TI - Homeopathy: a potent alternative. PMID- 12626170 TI - The Dalkon Shield saga. PMID- 12626171 TI - Size does matter. PMID- 12626173 TI - Newer progestogens. AB - OBJECTIVE: To review the literature on the most recent progestogens to be developed, to provide clinical comparisons with older progestogens and to look at the potential of products not yet marketed. DATA SOURCES: Searches of Medline and Popline together with requests for bibliographies from the Population Council, Wyeth-Ayerst Research and Schering Health Care. STUDY SELECTION: Information from technical papers was used to ascertain the metabolic characteristics and receptor binding affinities of the compounds. Previous reviews were scrutinised in order to make comparisons with older compounds. Any available trials were examined to ascertain efficacy, bleeding patterns and tolerability, more weight being given to comparative trials. DISCUSSION: Five progestogens have been developed in the last decade. They are all devoid of androgenic activity; some have antiandrogenic activity. Combined oral contraceptive (COC) pills containing dienogest and drospirenone are already marketed. Nomegestrol and nestorone have been extensively studied as subdermal implants. CONCLUSIONS: Newer progestogens used in combination with oestrogen behave very similarly to existing products. Progestogen-only products using new progestogens have potential for significantly better tolerability due to their lack of androgenic activity. PMID- 12626174 TI - Who has chlamydia? The prevalence of genital tract Chlamydia trachomatis within Portsmouth and South East Hampshire, UK. AB - OBJECTIVE: To determine the prevalence of genital tract Chlamydia trachomatis infection in women and men attending different health care settings in Portsmouth and South East Hampshire. DESIGN: Prospective, opportunistic screening. SETTING: Multiple health care sites. PARTICIPANTS: Consenting sexually active women and men. INTERVENTION: A urine sample was tested for Chlamydia trachomatis and positive patients were offered treatment and partner notification. MAIN OUTCOME MEASURES: The presence or absence of chlamydia infection according to age, gender, health care setting and reason for attendance. RESULTS: A total of 14,756 samples were tested giving an overall prevalence of 9.6%. The prevalence was significantly higher in women attending for a termination of pregnancy, antenatal care, women and men attending genitourinary medicine and in those with genital tract symptoms. The prevalence was different for men and women at different ages. CONCLUSION: The prevalence of genital Chlamydia trachomatis infection was high but differed at various health care settings and by reason for attendance. PMID- 12626175 TI - Intrauterine techniques: contentious or consensus opinion? AB - CONTEXT: Insertion of intrauterine devices (IUDs) is a routine procedure in Contraception & Sexual Health (C&SH) Service clinics. Techniques for IUD insertion vary between practitioners. OBJECTIVE: To describe the preferred approach to various aspects of IUD provision of experienced doctors working in three large, teaching C&SH Services, including policies on screening for chlamydia, antibiotic prophylaxis, use of tenaculae, use of analgesia/anaesthesia and use of assistants at the time of IUD insertion. DESIGN: An anonymous questionnaire to all doctors working in three neighbouring services. SETTING: Three community C&SH Services in Hampshire seeing in total approximately 92,000 patients each year. PARTICIPANTS: Doctors working regularly in target C&SH Services. RESULTS: A total of 94% of doctors cleanse the cervix prior to IUD insertion, 96% test for chlamydia before fitting an emergency coil and 18.5% always prescribe prophylactic antibiotics. For routine IUD insertions, 50% of doctors always screen for chlamydia prior to fitting the device. A total of 86% of doctors always stabilise the cervix with an Allis or similar instrument, with 14% reporting using an Allis 'sometimes' or 'rarely/never'. Instillagel was the most commonly used method of anaesthesia. A total of 75% of doctors have an assistant present for every insertion, eight doctors 'sometimes', and one 'rarely/never'. DISCUSSION: Arguments for and against each area of contention are discussed, and evidence reviewed. CONCLUSION: Practice varies between practitioners, and doctors training in intrauterine techniques may be given conflicting advice. All clinicians should be able to justify their practice on clinical grounds and audit outcomes. PMID- 12626176 TI - Homeopathic treatment for premenstrual symptoms. AB - Premenstrual syndrome (PMS) and premenstrual dysphoric disorder (PMDD) are well documented disorders causing significant morbidity in the female population. Treatments prescribed do not necessarily reflect proven clinical effectiveness. A recent systematic review from the Exeter Department of Complementary Medicine failed to endorse complementary therapies as a whole for treatment of PMS. However, a recent randomised controlled trial of homeopathic treatment for PMS confirms the clinical experience of homeopathic physicians that homeopathy is helpful in PMS. PMID- 12626177 TI - Can the Internet be used to improve sexual health awareness in web-wise young people? AB - OBJECTIVES: To assess Internet use amongst young people to determine whether it would be a practical way to provide sex education and information. METHODS: Year 10 students (aged 14-15 years) from North Nottinghamshire schools were asked to participate in focus groups to discuss the Internet. A series of predefined questions were directed to the whole group to generate debate. Areas explored included: Internet access and site; frequency and purpose of Internet use; websites visited; ideas for a genitourinary medicine (GUM) website. Responses were recorded by a hand count or as individual verbal responses. RESULTS: Thirteen focus groups were held involving 287 students of approximately equal sex distribution. All had access to Internet facilities at school and 224 (78.0%) had access elsewhere. Access was at least once a week by 178 (62.0%) mostly for e mail, games, chatlines and homework. No one accessed for health information. One hundred and seventy-nine (62.4%) participants said they would use a GUM website. A 'question line' where they could e-mail questions to a health care professional was of interest to 202 (70.4%) participants. CONCLUSIONS: The Internet would be a practical and accessible way of delivering sexual health education to young people, particularly if it is incorporated into activities and websites they enjoy. PMID- 12626179 TI - Experience of reversal of sterilisation at Glasgow Royal Infirmary. AB - OBJECTIVE: To review experience at the Glasgow Royal Infirmary with regard to women who underwent reversal of sterilisation, to obtain a profile of these women and to evaluate the procedure itself in terms of safety and restoration of fertility. DESIGN: A retrospective study based on case records from the Glasgow Royal Infirmary and from the Glasgow Royal Maternity Hospital, both of which share the same catchment area. PARTICIPANTS: Eighty-five women underwent reversal of sterilisation between 1 January 1994 and 31 December 1998 at the Glasgow Royal Infirmary. Follow-up at the Glasgow Royal Maternity identified 43/85 women. METHODS: Patients were identified from relevant theatre log books and from records at the maternity hospital until December 2000 and their case notes reviewed. RESULTS: Having a new partner was responsible for 90% of requests. Median age at request for reversal was 34 years and most had been sterilised before the age of 30 years. There were few postoperative complications. Pregnancy occurred in at least 43% of women. CONCLUSION: Reversal of sterilisation is a safe and effective method of restoring fertility. The actual incidence of pregnancy after reversal is likely to be higher than the 43.5% recorded due to difficulties in achieving 100% follow-up. PMID- 12626178 TI - Undiagnosed HIV in women attending a sexual health clinic for non-barrier methods of contraception. Two case studies. AB - Approximately one-third of human immunodeficiency virus (HIV) cases in the UK remain undiagnosed. More than half of new diagnoses in the last 2 years have been acquired through heterosexual sexual intercourse. Two cases of women with identified risk factors for HIV infection, who presented for contraception and were later found to be HIV-positive, are described. It is suggested that providers of contraception are well placed to identify clients at high risk of HIV and to recommend HIV testing. PMID- 12626180 TI - Simulators for intimate examination training in the developing world. AB - CONTEXT: Family planning clinic in urban India. OBJECTIVE: To assess the usefulness of a pelvic simulator for intimate examination and intrauterine device insertion training, and for enhancing reproductive education for women. DESIGN: Observational study using evaluation forms completed by staff, and focus group interviews with women and girls participating in sexual health education. Content analysis of interview material. RESULTS: Staff being trained in intimate examination found the simulator to be useful in increasing their level of skill. Participants in sexual health education reported an improved range and depth of knowledge following demonstrations with the simulator. CONCLUSION: Where working and training conditions are crowded and lacking in privacy, inappropriate examination practices are likely to be propagated. Training for intimate examinations can be enhanced by the use of pelvic simulators. PMID- 12626181 TI - Provider resistance to advance provision of oral contraceptives in Africa. AB - CONTEXT: In Africa, many new family planning clients are not menstruating at the time they present for services. Where pregnancy tests are unavailable, clients are often denied their method of choice and sent home to await menses. For pill clients, one obvious solution is 'advance provision' of oral contraceptives for later use. However, this practice is rare in Africa. OBJECTIVE: To assess the level of provider resistance to advance provision of oral contraceptives. DESIGN: We added questions about advance provision of pills to five provider surveys in three African countries. We also used simulated clients in Ghana to assess provider resistance to the practice. RESULTS: In Kenya, only 16% of providers thought it safe to give women oral contraceptives to be started at a later date. In Ghana and Senegal, fewer than 5% of providers mentioned advance provision as a way to manage non-menstruating pill clients. CONCLUSION: Training programmes and service delivery guidelines in developing countries should provide for advance provision of pills to appropriate clients PMID- 12626182 TI - An innovative approach to the assessment of laypersons' knowledge of contraceptive methods, followed by an educational intervention. AB - OBJECTIVES: To assess the knowledge of laypersons about contraceptive methods and to pilot an educational strategy. DESIGN: Cross-sectional survey using an anonymous questionnaire followed by an educational intervention. PARTICIPANTS: A total of 167 respondents to 400 invitations. METHOD: A questionnaire with 10 questions about contraceptive methods was answered before a meeting between laypersons and health professionals. Small groups facilitated by a health professional discussed the answers. One representative from each group presented the conclusions. At the end of the discussion session, gynaecologists gave lectures about contraceptive methods. RESULTS: Of the 167 people who attended, only 65 (39%) individuals handed over their anonymous answers for analysis. Those who did knew about the existence of contraceptive methods, but most of them showed incorrect or little knowledge about mechanism of action or risks. CONCLUSION: Embarrassment led many participants not to hand in their questionnaires but the results were evidence of the need for educational interventions. Lessons learnt from this experience are discussed. PMID- 12626196 TI - Left ventricular assist devices as alternative to heart transplant. When will they replace allogenic heart transplants? PMID- 12626183 TI - What constitutes 'unprotected sex'? PMID- 12626197 TI - Chlamydia pneumoniae and aortic aneurysms. PMID- 12626199 TI - Declining coronary heart disease mortality in Iceland: contribution by incidence, recurrence and case fatality rate. AB - OBJECTIVE: To analyse to what extent the recent decline in coronary heart disease mortality in Iceland is due to changes in incidence, recurrence and case fatality rates. DESIGN: A countrywide registration of myocardial infarction (MI) in people aged 25-74 was performed in Iceland during 1981-1999 according to the MONICA protocol. Possible cases were found by review of all hospital discharge records, autopsy records and death certificates. RESULTS: MI death rate declined by 63% in males and 51% in females, most in the youngest age groups in men (86%) and least in the oldest (49%). In women there was not a significant difference in age groups. Overall the age-adjusted reduction in MI death rate was 55.4% in both sexes combined; of this 23.1% was due to incidence reduction, 22.8% to recurrence reduction and 11.6% to case fatality reduction. In the youngest age groups the decline in incidence contributed most to the decline in MI death rate (62% in men and 71% in women), but thereafter the decline in case fatality in men. In the older age groups decline in recurrence rate has greater weight. CONCLUSION: The recent decline in MI mortality under the age of 75 years in Iceland is due to reduction in incidence and recurrence rate by about 40% each and to reduction in case fatality by 20%. PMID- 12626198 TI - Chlamydia pneumoniae in patients undergoing surgery for thoracic aortic disease. AB - OBJECTIVE: To investigate if Chlamydia pneumoniae is present in the wall of the thoracic aorta in patients operated on for aneurysm or aortic dissection. DESIGN: Consecutive patients undergoing surgery for thoracic aortic aneurysm (TAA, 32 patients) and for aortic dissection (6 patients) were included in this prospective study. Tissue samples from the aorta were analysed for the presence of C. pneumoniae by polymerase chain reaction (PCR), histopathology, immunohistochemistry and in one aortic tissue sample C. pneumoniae was verified by electron microscopy and immunogold labelling technique. Cultured Hep 2 cells infected with C. pneumoniae were used as a positive control for electron microscopy. Sera for microimmunofluorescence were obtained in 36/38 and throat swabs for C. pneumoniae PCR in 17/38 patients. RESULTS: Chlamydia pneumoniae was detected by PCR in 4 of 32 TAA tissue samples (12%) and in 0 of 6 patients operated on for aortic dissection. Chlamydia pneumoniae inclusion bodies in one of the PCR positive tissue samples were verified by electron microscopy. IgG antibodies to C. pneumoniae were present in 17/31 (55%) and IgA in 15/31 (48%) of the TAA patients and in none of five tested patients with dissection. None of the tested throat swabs was positive. CONCLUSION: In this study we report the presence of C. pneumoniae by PCR and electron microscopy in the wall of TAA. A high prevalence of serum IgA antibodies to C. pneumoniae was found in TAA patients. In contrast no signs of C. pneumoniae were detected in patients with thoracic aortic dissection. PMID- 12626200 TI - On the allocation of cardiac allografts from blood group-O donors. AB - An organ allocation policy, in which hearts from blood group-O donors are used to transplant recipients with other blood groups (ABO-compatible, non-identical transplantations), may affect blood group-O patients on the waiting list. We investigated how blood group affiliation influences potential recipients on the waiting list. In the case of patients with blood group O, fewer patients were transplanted, waiting list mortality was higher and waiting time to transplantation was longer. Patients with blood group O awaiting cardiac transplantation are affected considerably by an organ allocation policy in which ABO-compatible, non-identical transplantations are performed. PMID- 12626201 TI - Soluble adhesion molecules in coronary surgery and cardiopulmonary bypass with pump prime aprotinin. AB - OBJECTIVE: The purpose of the present study was to establish whether pump prime aprotinin could influence soluble adhesion molecules in patients undergoing elective coronary artery bypass surgery. DESIGN: Thirty patients admitted for first-time elective coronary artery bypass surgery were randomized into control or aprotinin groups. Patients in the aprotinin group received 280 mg of aprotinin in the pump prime. Plasma levels of soluble adhesion molecules were analyzed perioperatively. RESULTS: There were no significant changes in plasma sE-selectin after the operation in either group. Plasma sP-selectin increased significantly up to 20 h after reperfusion to the myocardium. Plasma sICAM-1 decreased in the early stage after cardiopulmonary bypass (CPB), then recovered at 4 h after reperfusion and a significant increase in sICAM-1 was observed 20 h later. There were no significant differences between the groups in postoperative changes in sP selectin (p = 0.21) and sICAM-1 (p = 0.91). CONCLUSION: Pump prime aprotinin did not influence plasma levels of E-selectin, P-selectin and ICAM-1 in the present patients. The present results do not support the concept of an anti-inflammatory effect of pump prime aprotinin. PMID- 12626202 TI - Cell adhesion and tissue factor upregulation in oxygenators used during coronary artery bypass grafting are modified by the Corline Heparin Surface. AB - OBJECTIVE: Cardiopulmonary bypass (CPB) is associated with inflammatory response and activation of coagulation. We investigated the influence of a new heparin surface on the activation of cells retrieved from oxygenators used during coronary artery bypass grafting (CABG). DESIGN: Sixty patients undergoing CABG with CPB were randomly assigned to either uncoated or completely Corline Heparin Surface (CHS)-coated circuits with one of three different levels of systemic heparin: standard, high or low. At end of surgery adhered cells were retrieved from the oxygenators and cell count, tissue factor (TF)- and CD11b-expression on monocytes and monocytic TFmRNA were analysed. RESULTS: The heparin coating of the oxygenator prevented adhesion of granulocytes, monocytes and platelets. TF expression on monocytes from the oxygenators was significantly higher than on circulating cells in all groups. Monocytes from the uncoated oxygenators showed low levels of TF-expression with high levels of TFmRNA. The coated group with high level of heparin showed higher surface-expression of TF with low levels of TFmRNA. CONCLUSION: The CHS was most biocompatible with the standard level of heparin used during CABG whereas elevation of systemic heparin rather increased the activation and TF upregulation in monocytes from oxygenators. PMID- 12626203 TI - Release patterns of pregnancy associated plasma protein A (PAPP-A) in patients with acute coronary syndromes. AB - OBJECTIVE: Pregnancy associated plasma protein A (PAPP-A) has recently been shown to be associated with acute coronary syndromes (ACS). The goal of this study was to investigate its release patterns in patients with ACS. DESIGN: PAPP-A concentrations in plasma samples serially collected after admissions from 15 patients with ACS were measured. The levels of PAPP-A were compared with a reference range determined from 80 normal subjects. The associations between PAPP A and myoglobin (Mb), C-reactive protein (CRP), fatty-acid-binding protein (FABP) and creatine kinase MB (CK-MB) were determined. RESULTS: Various release patterns were observed with 2-10-fold changes of PAPP-A in the different patients. Increases in PAPP-A levels above the reference range could appear early at 2 h or late at 30 h after onset of chest pain. Only in 4 of the 15 cases were significantly elevated PAPP-A levels detected before 6 h after onset. Elevations early after admission showed rapid decline whereas later elevations were more persistent. No associations between PAPP-A and Mb, CRP, FABP and CK-MB were found. However, a weak but significant association to cardiac troponin I (cTn I) was found. CONCLUSION: PAPP-A is an additional marker for ACS, but does not seem to be a useful early marker for acute myocardial infarction (AMI). The possible clinical utility of PAPP-A calls for extensive studies of chest pain patients using serial sampling combined with short- and long-term outcome studies. PMID- 12626204 TI - No effect of cyclooxygenase inhibition on plaque size in atherosclerosis-prone mice. AB - OBJECTIVE: To study the role of cyclooxygenase (COX) inhibition on the development of advanced atherosclerosis in apolipoprotein E-deficient (apoE(-/-)) mice. DESIGN: Sixty apoE(-/-) mice were divided into three groups: a control group, a group fed standard mouse chow supplemented with 0.0067% (wt/wt) MF Tricyclic (selective COX-2 inhibitor), and a group fed the diet supplemented with 0.0134% (wt/wt) sulindac (non-selective COX inhibitor). Four months later, the mice were killed and the atherosclerotic plaque area in the aortic root was measured. RESULTS: Mean body weights did not differ at any time. The MF Tricyclic and sulindac groups had drug plasma levels of 1.31 +/- 0.11 and 0.84 +/- 0.23 micro g/ml, respectively. Plasma total cholesterol and triglyceride values were similar in all three groups. A small difference in plasma levels of high-density lipoprotein cholesterol was found between the groups (p = 0.03). Advanced atherosclerotic plaques were present in mice from all three groups, but there was no difference in mean plaque size between the groups (p = 0.9). CONCLUSION: Neither selective COX-2 nor non-selective COX inhibition influenced the development of advanced atherosclerosis in apoE(-/-) mice. PMID- 12626206 TI - A comparison of spiral computed tomography and latex agglutination D-dimer assay in acute pulmonary embolism using pulmonary arteriography as gold standard. AB - OBJECTIVE: To compare the diagnostic accuracy of contrast medium enhanced spiral computed tomography of the pulmonary arteries (s-CTPA) and a latex agglutination D-dimer assay in patients with suspected acute pulmonary embolism (PE) by using pulmonary arteriography (PA) and clinical follow-up as reference method. DESIGN: Ninety hemodynamically stable patients with symptoms of acute pulmonary embolism were prospectively evaluated with s-CTPA and pulmonary arteriography (PA) within 24 h from admission. Plasma D-dimer levels on admittance were analyzed using a rapid latex agglutination D-dimer assay. The outcome of D-dimer concentrations in plasma below 0.25 and 0.5 mg/l was studied. RESULTS: All PA and s-CTPA investigations were regarded as of acceptable diagnostic quality in a consensus reading. Thirty-three patients had a positive PA (37%). Three patients had false negative and two patients had false positive s-CTPA findings. s-CTPA had 91% sensitivity, 96% specificity, 94% positive predictive value (PPV) and 95% negative predictive value (NPV). The sensitivity and specificity for D-dimer below 0.5 mg/l were 79 and 88%, respectively. The PPV and NPV were 81 and 87%. If a cut-off level of 0.25 mg/l was used the corresponding figures were 91, 65, 63 and 92%. CONCLUSION: s-CTPA has a higher sensitivity and specificity than latex agglutination D-dimer. A cut-off level of 0.25 mg/l can be used as screening method, but s-CTPA must be performed to exclude false positive cases. PMID- 12626205 TI - The kinase insert domain-containing receptor (KDR) is regulated by shear stress. AB - OBJECTIVE: Intimal hyperplasia develops in areas with low shear stress. The aim of the present study was to investigate if the mRNA expression of vascular endothelial growth factor (VEGF), Fms-like tyrosine kinase-1 receptor (Flt-1) and kinase insert domain-containing receptor (KDR) is regulated by shear stress. DESIGN: Endothelial cells from human umbilical veins were in an in vitro system subjected to different levels of shear stress during 1 and 12 h. The mRNA expression of VEGF, Flt-1 and KDR was measured with RT-PCR. eNOS served as positive control and actin as housekeeping gene. RESULTS: The KDR expression was isolated upregulated 3-4 times after 12 h exposure to high shear stress. CONCLUSION: The genetic expression of KDR is upregulated by shear stress and this response is time dependent. PMID- 12626207 TI - Leg wound closure after saphenous vein harvesting in patients undergoing coronary artery bypass grafting: a prospective randomized study comparing intracutaneous, transcutaneous and zipper techniques. AB - OBJECTIVE: Two prospective randomized studies were undertaken to compare different suture closure techniques with respect to postoperative wound infection rates and cosmetic results after saphenous vein harvesting in patients undergoing coronary artery bypass surgery. DESIGN: A total of 166 patients were included in the first study, in which 85 had their leg wounds closed with transcutaneous and 81 with intracutaneous suture. In the second study, 168 patients were selected to a non-invasive surgical zipper (n = 78) or intracutaneous suture (n = 90). RESULTS: In the first study the overall infection rate was 20.5%, 17.6% in the transcutaneous group compared with 23.5% in the intracutaneous group (p = 0.35). In the second study the infection rate was 19.3%, 15.3% in the zipper group vs 23.3% in the intracutaneous group (p = 0.20). On a cosmetic scale from 1 to 10, an average score of 8.0 was obtained in the percutaneous (p.c.) group vs 8.3 in the intracutaneous (i.c.) group (p = 0.35), and 9.0 in the zipper group vs 8.4 in the i.c. group (p = 0.003). CONCLUSION: The incidence of leg wound infection after saphenous vein harvesting in coronary artery bypass graft surgery is high. The zipper closing method may give a lower infection rate and a better cosmetic result compared with the intracutaneous suture. PMID- 12626208 TI - The end of WISDOM. PMID- 12626209 TI - WISDOM: history and early demise - was it inevitable? AB - In 1989, the UK Medical Research Council (MRC) agreed that, if feasible, a randomized controlled trial to assess the long-term risks and benefits of hormone replacement therapy (HRT) was a priority. Feasibility work began in 1990 and demonstrated that a large-scale multicenter trial was possible. An application for funding for a main trial was submitted to MRC in 1993 and, after extensive review, funding was released in late 1996. Set-up work for the trial - the Women's International Study of long Duration Oestrogen after Menopause (WISDOM) - began in 1997 with recruitment in 1999. In October 2002, following the early discontinuation of one arm of the US Women's Health Initiative HRT trial, the MRC decided to stop the WISDOM trial. This article, by the principal UK investigators of WISDOM, sets out the background and history of the trial. PMID- 12626211 TI - A tale of two studies: reflections on WHI and WISDOM. PMID- 12626210 TI - Cessation of the WHI and WISDOM trials: a New Zealand perspective. PMID- 12626212 TI - Combined hormone replacement therapy and risk of breast cancer in a French cohort study of 3175 women. AB - The largest-to-date randomized trial (Women's Health Initiative) comparing the effects of hormone replacement therapy (HRT) and a placebo concluded that the continuous use of an oral combination of conjugated equine estrogens (CEE) and medroxy-progesterone acetate (MPA) increases the risk of breast cancer. This conclusion may not apply to women taking other estrogen and progestin formulations, as suggested by discrepancies in the findings of in vitro studies, epidemiological surveys and, mostly, in vivo studies of human breast epithelial cell proliferation showing opposite effects of HRT combining CEE plus MPA or estradiol plus progesterone. To evaluate the risk of breast cancer associated with the use of the latter combination, commonly prescribed in France, a cohort including 3175 postmenopausal women was followed for a mean of 8.9 years (28 367 woman-years). In total, 1739 (55%) of these women were users of one type of estrogen replacement with systemic effect during at least 12 months, any time after the menopause, and were classified as HRT users. Among them, 83% were receiving exclusively or mostly a combination of a transdermal estradiol gel and a progestin other than MPA. Some 105 cases of breast cancer occurred during the follow-up period, corresponding to a mean of 37 new cases per 10 000 women/year. Using multivariate analysis adjusted for the calendar period of treatment, date of birth and age at menopause, we were unable to detect an increase in the relative risk (RR) of breast cancer (RR 0.98, 95% confidence interval (CI): 0.65 1.5) in the HRT users. The RR of breast cancer per year of use of HRT was 1.005 (95% CI 0.97-1.05). These results do not justify early interruption of such a type of HRT, which is beneficial for quality of life, prevention of bone loss and cardiovascular risk profile, without the activation of coagulation and inflammatory protein synthesis measured in users of oral estrogens. PMID- 12626214 TI - Hormone replacement therapy use over a decade in an Australian population. AB - OBJECTIVES: To describe the prevalence of hormone replacement therapy (HRT) during the 1990s in an Australian population and the length of time this therapy has been used. Design Data were obtained from five biennial representative population face-to-face interview surveys of the South Australian population from 1991 to 2000, and an additional telephone survey in 2001. The main surveys used consistent methods and quality control procedures. PARTICIPANTS: Over 3000 South Australian adults were interviewed in their own homes by trained health interviewers in each of the biennial surveys. RESULTS: In 2000, 28% of women aged over 50 were currently using HRT, and ever-use of HRT was 43%. Highest use of HRT was in the 55-59-year age group, where current use was 47% and ever-use was 70%. Among women 55 years and older there was a statistically significant increase in the prevalence of HRT use between 1991 and 2000. Mean length of use in women aged over 55 years increased from 5.4 years in 1991 to 9.6 years in 2000. CONCLUSIONS: Consistent surveillance of the population has shown that HRT use has not increased before age 55, but has greatly increased thereafter. Long-term effects may now be occurring, and further results of current long-term randomized trials are awaited to confirm the overall risk/benefit ratio suggested by observational studies and early results from long-term trials. PMID- 12626213 TI - Differing effects of low-dose estrogen-progestin therapy and pravastatin in postmenopausal hypercholesterolemic women. AB - BACKGROUND: Most studies examining the potential cardioprotective effects of postmenopausal estrogen have been undertaken in healthy women, with doses that may not be appropriate for long-term intervention. New low-dose estrogen progestin regimens alleviate postmenopausal symptoms with a favorable side-effect profile; however, little is known of the impact of such regimens in women at increased risk of cardiovascular disease. Hence, we have evaluated the effects of low-dose oral estrogen-progestin therapy on serum lipoprotein lipids, brachial artery reactivity and fibrinogen in hypercholesterolemic postmenopausal women in direct comparison with the effects of pravastatin, a lipid-lowering agent known to reduce cardiovascular events in women. METHODS: In a randomized, double-blind, double-dummy, parallel trial, we studied the effects of continuous combined estrogen-progestin therapy (1 mg 17beta-estradiol with 500 micro g norethisterone acetate daily) or pravastatin (20 mg daily) in 72 postmenopausal women with fasting serum low-density lipoprotein (LDL) cholesterol levels greater than 124 mg/dl after an 8-week run-in diet, over a 24-week period. The primary end-point was percentage change in LDL cholesterol from baseline. RESULTS: The intention-to treat population comprised 65 women, mean age 59 +/- 6.3 years, and 29 in each group completed the trial. Diet alone reduced LDL cholesterol significantly in both treatment groups, in association with a reduction in weight during this period. Compared with respective baseline values, pravastatin decreased LDL cholesterol and total cholesterol to a greater extent than hormone therapy (p = 0.0001 and 0.003 for difference between treatments, respectively). High-density lipoprotein (HDL) cholesterol levels decreased with hormone therapy, but did not change with pravastatin (p = 0.01). Lipoprotein(a) decreased significantly with hormone therapy only (-14%, 95% confidence interval (CI) -21 to -6%, p = 0.01 for difference between groups). Brachial artery flow-mediated dilatation (FMD) was impaired at baseline, and this increased with hormone therapy (absolute mean change in artery diameter as percentage units 2.07, 95% CI 0.57-3.57, p = 0.009) versus no change with pravastatin (0.19, 95% CI -1.1 to 1.5, p = 0.78), with a near-significant difference between the two groups (p = 0.058). A significant correlation between improved brachial artery FMD and reduction in LDL cholesterol was not observed. Fibrinogen decreased significantly in both treatment groups with no difference between treatments. CONCLUSIONS: In postmenopausal hypercholesterolemic women, pravastatin and hormone therapy exhibited divergent effects. The former lowered total and LDL cholesterol more effectively, whereas hormone therapy lowered lipoprotein(a) significantly and improved brachial artery endothelium-dependent dilatation, independent of the reduction in LDL cholesterol. The modest increase in brachial artery FMD seen is consistent with hypercholesterolemia compromising endothelial integrity, and suggests that the important effect of estrogen on the endothelial microenvironment may be attenuated in women with endothelial dysfunction. PMID- 12626215 TI - Addition of testosterone to estrogen replacement therapy in oophorectomized women: effects on sexuality and well-being. AB - OBJECTIVE: To evaluate the effect of adding testosterone undecanoate 40 mg daily to estrogen replacement on sexual function, psychological well-being and self esteem in surgically postmenopausal women. METHODS: A letter of invitation to participate in the study was mailed to women who had undergone hysterectomy and bilateral oophorectomy for benign disorders during 1990-98. Fifty women, 45-60 years old, were consecutively recruited and randomly assigned to oral treatment with testosterone undecanoate 40 mg plus estradiol valerate 2 mg daily or placebo plus estradiol valerate 2 mg daily for 24 weeks. A double-blind design was chosen, with cross-over to the other regimen for another 24 weeks of treatment. Forty-four women completed the study. Outcome included scores on McCoy's sex scale questionnaire, the Psychological General Well-Being index and a self-esteem questionnaire, at baseline and after 24 weeks of either treatment. Serum concentrations of total testosterone, sex hormone binding globulin, free testosterone, dihydrotestosterone, androstenedione, estradiol, follicle stimulating hormone and luteinizing hormone were analyzed at baseline and after 24 weeks of both treatment regimens. RESULTS: After 24 weeks, both treatment regimens had significantly improved some of the sexual variables. The addition of testosterone had a significantly better effect on the sex variables 'enjoyment of sex', 'satisfaction with frequency of sexual activity' and 'interest in sex'. The total McCoy score was significantly increased by both treatments, but there was a stronger effect when testosterone was also given. Although both regimens improved psychological well-being and self-esteem, we found no significant differences between testosterone-estrogen or estrogen alone at 24 weeks. Serum levels of all androgens, with considerable individual variation, increased significantly from baseline after 24 weeks of testosterone-estrogen treatment. Supraphysiological levels were achieved in a significant proportion of the women. Increases in estradiol and sex hormone binding globulin were less marked when testosterone was also given. Both treatments reduced gonadotropin levels. CONCLUSIONS: The addition of testosterone undecanoate improved specific aspects of sexual function more than treatment with estrogen alone. Improvements in well-being and self esteem were similar for both treatments. If testosterone undecanoate 40 mg daily should be used for clinical treatment, regular monitoring of androgen serum levels is needed. PMID- 12626217 TI - Attitudes towards and use of dietary supplementation in a sample of postmenopausal women. AB - OBJECTIVES: To assess the reason why people take food supplements in addition to or in place of medication, as well as the perceived efficacy and risk of these supplements. METHODS: A 20-item, computer-assisted telephone questionnaire was administered to 442 women over 60 years of age in Hull, to assess the prevalence of use and perception of efficacy and safety of food supplements. RESULTS: Data were available for 411 women, of whom 269 (65.4%) were currently taking food supplements, 76 (18.5%) were previous users and 66 (16%) were never-users. There was no difference in terms of age, smoking, consumption of fruit and vegetables, life-style and attitude towards conventional medicine between current or past users of supplements and never-users. The majority of users (n = 229, 66%) thought that food supplements help maintain good health (p < 0.001). Overall long term (5 years) adherence was achieved in 244 cases of supplement uses (36%). Among users, 150 (43%) thought that supplements were very effective, and 171 (49.5%) women had noticed an improvement of various symptoms. Although the majority of users (n = 176, 51%) believed that supplements might have adverse effects, over 66% would not associate side-effects with the food supplement. Information regarding supplements was obtained primarily from the media, particularly magazine and newspaper articles (27%), with health professionals rarely being consulted (16%). CONCLUSIONS: Users believed supplements to be effective, although possibly with associated side-effects. However, they would not attribute adverse effects experienced to this form of treatment. Medical practitioners and health professionals are rarely informed or consulted, leaving potentially dangerous side-effects of food supplements to go unreported. PMID- 12626216 TI - Hysterectomy and ovarian function: levels of follicle stimulating hormone and incidence of menopausal symptoms are not affected by hysterectomy in women under age 45 years. AB - OBJECTIVE: To investigate levels of follicle stimulating hormone (FSH), as a measure of ovarian function and menopausal symptoms prior to and following hysterectomy in women under the age of 45 years. DESIGN: This was a prospective controlled study. SAMPLE: A total of 56 hysterectomy patients and 34 controls, up to the age of 42 years, were recruited. METHODS: Women undergoing hysterectomy were recruited prior to their operation and were matched, where possible, with controls for age, parity and smoking. Five blood samples were taken over 2 years, two before the operation (or at a 1-month interval in the control group) and then at 6 months, 1 year and 2 years. Questionnaires were completed at each sampling to assess symptoms associated with reduced estrogen secretion. RESULTS: No significant differences in FSH levels between patients and controls were found. Modest differences in FSH levels between different time-points were identified (p = 0.03), but this disappeared if age and smoking were included as covariates. For the questionnaire data, there were no significant differences between the two groups on the somatic and vasomotor scales, but for psychological symptoms the patients scored significantly higher than the controls (p = 0.007), particularly at the pre-operation time-point. CONCLUSION: The study found no evidence of compromise of ovarian function, as reflected in FSH levels, within 2 years of hysterectomy. Psychological symptoms were higher in women undergoing hysterectomy both before and after the operation. PMID- 12626218 TI - Comparative study to evaluate skin irritation and adhesion of Estradot and Climara in healthy postmenopausal women. AB - OBJECTIVES: To compare the local tolerability, adhesion and estradiol delivery of a 5-cm(2) transdermal patch (Estradot), Novartis Pharmaceuticals, Basel, Switzerland) and a 12.5-cm(2) patch (Climara, Berlex Laboratories, Wayne, NJ, USA). METHODS: This was an open-label, randomized, intrapatient, comparative study. One hundred healthy postmenopausal women applied the 50 micro g/day 5 cm(2) patch and the 12.5-cm(2) patch concurrently for 7 days; safety and tolerability were assessed. Twelve women continued to apply the 5-cm(2) and 12.5 cm(2) patches separately for 7 days in a two-way cross-over study, to investigate the reproducibility of estradiol delivery. RESULTS: The proportion of subjects with a clinically significant erythema score was higher with the 12.5-cm(2) patch. The 5-cm(2) patch had a significantly lower incidence of very slight erythema than the larger patch (21.4% vs. 32.3%; p = 0.0028). Overall erythema scores were 0.22 and 0.41, respectively. More 5-cm(2) patches had > 90% adherence than the 12.5-cm(2) patches (87.5% vs. 82.0%, not significant) and fewer became detached (0.5% vs. 3.0%). Both the patches showed reproducible delivery of estradiol. CONCLUSION: The 5-cm(2) patch was associated with less skin irritation and better adherence than the 12.5-cm(2) patch, although the majority of the differences were not significant. The 5-cm(2) patch was well tolerated and showed reproducible estradiol delivery, as did the 12.5-cm(2) patch. PMID- 12626220 TI - 'Executive summary of the Stages of Reproductive Aging Workshop (STRAW)': not less, but more confusion. PMID- 12626219 TI - Ten years of treatment with tibolone 2.5 mg daily: effects on bone loss in postmenopausal women. AB - OBJECTIVES: To assess the effects of tibolone 2.5 mg daily over a 10-year period on bone loss and other efficacy parameters in postmenopausal women, and to confirm its long-term safety and tolerability. METHODS: A 10-year, open, non randomized, prospective study was conducted in 110 postmenopausal women; 59 women received tibolone 2.5 mg daily and 51 were entered into an untreated control group. After 10 years, 31 women in the tibolone group (53%) and 26 (51%) in the control group remained in the study. RESULTS: After 10 years, tibolone had significantly (p < 0.0001) increased the mean bone mineral density in the lumbar spine and femoral neck by 4.8% and 3.7%, respectively, compared with baseline. The corresponding changes in the control group were -8.5% and -8.9% (p < 0.0001 vs. baseline), respectively. The incidence and intensity of hot flushes were reduced in the tibolone group and there were improvements in vaginal cytology parameters. Tibolone was well tolerated. CONCLUSION: Tibolone 2.5 mg daily maintains its beneficial effects on bone loss over 10 years, while remaining well tolerated. PMID- 12626221 TI - Perinatal mortality in triplet births in Japan: time trends and factors influencing mortality. AB - Perinatal mortality rates (PMRs) in triplets were analyzed using Japanese Vital Statistics during the period of 1980-1998. The total number of perinatal deaths in triplets was 1051. The PMR significantly decreased from 214 per 1000 births in 1980 to 39 in 1998, a reduction of 82%. PMRs in triplets were 11.1-fold higher in 1980 and 6.9-fold higher in 1998 than in singletons, indicating that PMRs improved more in triplets than in singletons during the last two decades in Japan. The PMR was the highest in the third-born, followed by the second- and the first-born triplets in each period. As for maternal age, the PMR was 1.5-3.7 times higher in the < 25 years of age group than the other age groups. Additionally, the PMR was the lowest for birthweight (BW) >or= 2000 g during the entire period. In addition, the PMR decreased with gestational age (GA) of up to 38-39 weeks and increased thereafter. The effects of BW on the PMR were stronger than the effects of GA. The proportion of perinatal deaths in triplets with extremely low BW (< 1000 g) was 74% in 1980-1989 and increased to 82% in 1990 1998. The declining PMR was unlikely to be due to the improvement in BW in triplets. It is likely that it was related to the improved medical management of triplets during the perinatal period and the first week of life. Information obtained in the present study may be useful in counseling pregnant woman about triplet births. PMID- 12626222 TI - A matched cohort comparison of the outcome of twin versus singleton pregnancies in Flanders, Belgium. AB - To compare perinatal outcome of singleton versus twin pregnancies a matched cohort study was performed in Flanders, Belgium. All twins delivered in the region of Flanders during 1998-1999 were compared to singletons, matched for gestational age, fetal sex and maternal parity, resulting in 4384 infants in each group. Above 32 weeks of gestation, birthweight was significantly lower in twins (2095 +/- 364 g versus 2315 +/- 523 g; p < 0.001, 95% confidence interval 193 to 246 g). Perinatal mortality was also significantly lower in twins (1.98% versus 1.26%; odds ratio for twins 0.63; 95% confidence interval 0.53-0.75; p < 0.001 ), this was mostly due to fetal and not to early neonatal mortality. Congenital malformations occurred less frequently in twins (2.5% versus 3.7%; odds ratio for twins 0.80, 95% confidence interval 0.69-0.92; p = 0.001). From gestational age of 32 weeks on, respiratory distress syndrome was less frequent in twins (6.7% versus 8.0%; odds ratio for twins 0.81; 95% confidence interval 0.68-0.97; p = 0.011 ). No significant differences were noted with regard to intraventricular haemorrhage, neonatal infections and retinopathy of prematurity. Although twins have a lower birthweight, their outcome is more favorable compared to singletons, when matched for gestational age. PMID- 12626223 TI - Newborn twin outcome predicted by maternal variables: differentiation by term and sex. AB - Predictive relations between maternal variables and outcome status for 199 pairs of newborn twins were evaluated. The maternal independent variables included pregravid weight, weight at end of pregnancy, weight gain, age, parity, height, socioeconomic status, years of education, and body mass index. The twin dependent variables included birthweight, birth length, 5-min Apgar score, number of days in an isolette, number of days in the hospital, weight at discharge, and chronological age at discharge. For stepwise multiple linear regression analyses, the newborn twins were grouped by term (fullterm/preterm) and sex (female/male). A combination of a larger number of independent variables acted as statistically significant predictors of twin outcome for the preterm twins than for the fullterm twins, and for the male twins than for the female twins. This was noted particularly for the anthropometric variables and for parity. Maternal weight gain, typically considered to be the critical variable related to infant birthweight, consistently had a significant outcome role for the preterm twins and male twins, but not for the fullterm twins or female twins. Demographic variables had a low rate of prediction for newborn outcome. This study has demonstrated that evaluating twins within groups separated by potential risk variables allows for a more accurate description of the influence of maternal variables on twin outcome than evaluating twins across groups as a single population. PMID- 12626224 TI - Retrospective determination of chorion type in twins using a simple questionnaire. AB - This study investigates the validity of retrospective determination of chorion type by asking the question to the mother about the number of placentas. In the "East Flanders Prospective Twin Survey" (EFPTS), accurate information on the placentation and zygosity of the multiples was collected prospectively. The mothers of 231 monozygotic (95 dichorionic and 136 monochorionic) twins and 255 dizygotic twins were asked to fill in a simple questionnaire regarding 1). the zygosity and 2). the number of placentas of their twins. The accuracy of the response to the question on "the number of placentas" was 60% for monozygotic twins and 37% for dizygotic twins. The accuracy of the response to the question on the zygosity of the twins was 93% for monozygotic and 95% for dizygotic twins. If the questionnaire was used for the determination of chorion type, a total of 31 monozygotic twins (13%) should have been assigned as dichorionic on the fact that there were two separate placentas. Of these, 10 (32%) are monochorionic and 12 (39%) were falsely reported as having two placentas. We conclude from these findings that this simple questionnaire method is unreliable for the retrospective determination of the chorion type. PMID- 12626225 TI - Zygosity diagnosis in the absence of genotypic data: an approach using latent class analysis. AB - For zygosity diagnosis in the absence of genotypic data, or in the recruitment phase of a twin study where only single twins from same-sex pairs are being screened, or to provide a test for sample duplication leading to the false identification of a dizygotic pair as monozygotic, the appropriate analysis of respondents' answers to questions about zygosity is critical. Using data from a young adult Australian twin cohort (N = 2094 complete pairs and 519 singleton twins from same-sex pairs with complete responses to all zygosity items), we show that application of latent class analysis (LCA), fitting a 2-class model, yields results that show good concordance with traditional methods of zygosity diagnosis, but with certain important advantages. These include the ability, in many cases, to assign zygosity with specified probability on the basis of responses of a single informant (advantageous when one zygosity type is being oversampled); and the ability to quantify the probability of misassignment of zygosity, allowing prioritization of cases for genotyping as well as identification of cases of probable laboratory error. Out of 242 twins (from 121 like-sex pairs) where genotypic data were available for zygosity confirmation, only a single case was identified of incorrect zygosity assignment by the latent class algorithm. Zygosity assignment for that single case was identified by the LCA as uncertain (probability of being a monozygotic twin only 76%), and the co twin's responses clearly identified the pair as dizygotic (probability of being dizygotic 100%). In the absence of genotypic data, or as a safeguard against sample duplication, application of LCA for zygosity assignment or confirmation is strongly recommended. PMID- 12626226 TI - Serum inhibin A and B concentrations during the menstrual cycle in mothers of spontaneous dizygotic twins. AB - Dizygotic twinning in humans is influenced by genetic factors suggesting inherited variation affects follicle development and predisposes to double ovulations. In a previous study, we conducted a detailed examination of follicle development and variation in hormone concentrations during the menstrual cycle in mothers of DZ twins (MODZT) compared with an age-matched control group of mothers of singletons. We did not detect differences in FSH concentrations between mothers of twins and mothers of singletons. Serum inhibin concentrations were measured by a radioimmunoassay that did not distinguish between dimeric inhibin A and B forms and free inhibin alpha subunit. We therefore analyzed the samples from this study with specific assays to determine whether concentrations of inhibin A and B were different between MODZT and controls and therefore contribute to the twinning phenotype. There were no significant differences between MODZT with single ovulations and control women in inhibin A and B concentrations during the cycle, including the critical period for the selection of the dominant follicle. These data suggest that the genetic cause of twinning is not associated with changes in FSH concentrations or recognised feedback mechanisms regulating FSH release. PMID- 12626227 TI - Breastfeeding rights of multiple birth families and guidelines for health professionals. AB - Increasing numbers of women wish to breastfeed their multiple birth children. Breastfeeding of preterm and fullterm multiple birth infants is complex and demanding for the families and presents distinct challenges for health professionals. Families require sustained assistance from health care providers who are encouraging, knowledgeable, skilled, and committed to the breastfeeding of multiple birth children. Seven breastfeeding rights of multiple birth families are presented for the continuum of pregnancy to early childhood and are in accordance with the Declaration of Rights and Statement of Needs of Twins and Higher Order Multiples (Council of Multiple Birth Organizations of the International Society for Twin Studies, 1995). Guidelines for each of the rights have been developed to assist health professionals provide "best practices" in community and hospital settings. The guidelines are based on the existing body of breastfeeding of multiples' research, empirical findings, and consultations with parents and care providers with experience and/or expertise in breastfeeding multiples. The rights and guidelines suggest direction for providing assistance, implementing programs and services, conducting research, and evaluating the effectiveness of multiples-specific breastfeeding care during the prenatal, infancy, and toddlerhood periods. PMID- 12626228 TI - Small for gestational age as a predictor of behavioral and learning problems in twins. AB - The study examines the effects of being born small for gestational age (SGA) on rates of behavioral problems in twins and siblings, utilising data from the Australian Twin Study of disruptive behavior disorders in twins and their siblings. Participants were 3944 twins and their siblings who were assessed at two intervals three years apart. At the first assessment (1991), they ranged between 4 and 12 years of age. Items assessing Attention Deficit/Hyperactivity Disorder were based on DSM-III-R criteria (Time 1) and DSM-IV criteria (Time 2). Other measures included history of speech and reading therapy, demographic information and obstetric and neonatal history. Results indicated that both male and female twins, who were extremely growth restricted (small for gestational age up to the third percentile-WGA3) showed more inattention, and poorer speech and reading scores. The effects were greater for males. Male twins who were small for gestation age, up to the 10th percentile, were more likely to have a DSM-IV diagnosis of Inattention. Implications of these results included WGA3 male twins being at a "triple disadvantage" for subsequent behavioral and learning problems, in that being male, being a twin and being small for gestational age are all significant factors. Recommendations are made for early intervention for low birthweight male twins. The study is consistent with recent follow-up studies of very-low-birthweight singletons, indicating male disadvantage in cognitive outcome. While there is some genetic component to SGA, it does constitute a potentially major contribution to common environmental effects that must be considered in twin-based genetic analyses. PMID- 12626229 TI - Stress and coping among mothers of twins in rural southwestern Nigeria. AB - High rates of twinning have been reported in Africa. This study sought to learn whether mothers of twins experience different stress levels than those who have only singletons. Both twin mothers (101) and singleton mothers (101) were interviewed. Using a scale of 15 stressors the researchers found that the mean stress score was significantly higher for twin mothers than singleton mothers. Higher stress scores were also associated with parity and perceptions of problems in caring for children and inversely related to perceived levels of social support. Confronting stressors or simply accepting them were the two main ways that mothers coped with their problems. PMID- 12626230 TI - Osteoarthritis of the hands, hips and knees in an Australian twin sample- evidence of association with the aggrecan VNTR polymorphism. AB - Age-related changes in the composition of the cartilage matrix may be associated with the development of osteoarthritis, a relatively late-onset disease characterised by the destruction of joint cartilage. In order to investigate whether differences in the VNTR polymorphic region of aggrecan affect cartilage functionality and therefore the development of osteoarthritis, we examined the aggrecan polymorphic genotypes of a sample of 134 Australian twins aged over 50 (including 34 monozygotic and 27 dizygotic twin pairs). Clinical measures of hand, hip and knee osteoarthritis, as well as self-reported bone and joint pain, were tested for association with the aggrecan polymorphism. The results were consistent with either a deleterious effect of allele 27, or a protective effect of alleles 25 and 28, providing some additional evidence for an association between the aggrecan VNTR polymorphism and osteoarthritis of the hands, hips and knees. PMID- 12626231 TI - Challenges in recruiting older twins for the Sri Lankan twin registry. AB - The National Twin Registry of Sri Lanka was established in 1997 as a volunteer register. To extend it to a population-based register, we examined the effectiveness of tracing older twins by inspecting birth records and recruiting them by postal invitation and in-person contact. Birth records at a divisional secretariat reported from 2 maternity hospitals between the years of 1954-1970 were scrutinised to identify a random sample of twins. These hospitals had the highest twin delivery rates for the whole country. We identified 620 twins and a questionnaire was mailed to them. Research assistants visited a cohort of non respondents (71) in the postal survey. These 620 twins were identified after perusing 20700 birth records. The twinning rate was estimated at 29.95 ([620/20700] x 1000) twins per 1000 registered births (CI 27.63-32.27). In the postal survey, 37 (12%) responded and 62 letters were returned (20%). Both twins were still alive in 20 pairs, one was still alive in 15 pairs, and both twins were dead in 2 pairs. During field visits, 42 (59.2%) addresses were located. Information was available on 16 twin pairs. Both twins were alive in 8 pairs, one each in 4 pairs, and both were dead in 4 pairs and at least one twin was traced in 10 pairs (14%). Both the postal and the field survey gave a low yield. This finding is different from tracing younger twins born between 1985-1997 by using the same methods. Migration, urbanization and development in the country might have affected tracing older twins from the birth record addresses, which were decades old. PMID- 12626233 TI - The stratosphere. AB - The stratosphere is that part of the atmosphere which lies between ca. 10 and 50 km above the surface of the Earth and which contains the ozone layer. It is the seat of much interesting behaviour in terms of dynamics, radiation and chemistry, now revealed in detail by observations from modern space instruments, but still not completely understood. Other planetary atmospheres exhibit stratospheric behaviour which in some ways resembles, and in others contrasts sharply with, that of the Earth. In reviewing these topics, this paper describes some key problems that will be addressed by new measurements from space in the near future. PMID- 12626234 TI - The GAMBLE project: a fresh approach to satellite remote sensing of the sea. AB - Global Altimeter Measurements by Leading Europeans (GAMBLE) is a project, funded by the European Commission, which aims to mobilize the community of specialists in radar altimetry within Europe. The main objective is to investigate how to maximize the synergy between satellite altimetry missions before launch. This makes GAMBLE unique in the annals of planning future satellite missions over the oceans and represents a notable first for Europe. PMID- 12626235 TI - Measurements of the offshore wave climate around the British Isles by satellite altimeter. AB - Satellite altimetry gives a new perspective on ocean wave climate. Measurements around the British Isles show a strong seasonality, with exceptionally large average wave heights to the west and north of the British Isles in the winter. Furthermore, the interannual variability of winter wave climate is very high. Most of this variability can be described by a strong linear dependence on the North Atlantic Oscillation (NAO) index. This relationship may largely explain observations of increasing wave heights in the northeastern Atlantic and northern North Sea during the latter decades of the 20th century, coincident with a long term rise in the NAO. PMID- 12626239 TI - Satellite observations of the Agulhas Current system. AB - The Agulhas Current system is a complex interplay of currents and eddies with the bathymetry. Components such as the East Madagascar Retroflection and the Agulhas Return Current evolve significantly over a month, and they are thus not adequately resolved by infrequent research-ship cruises. This paper contrasts the abilities of three different spaceborne sensors for monitoring these complex regimes. A key parameter is sea-surface temperature, measured by both infrared and microwave radiometers. Ocean colour observations of chlorophyll can also be used to distinguish between water masses. PMID- 12626236 TI - Assimilating satellite ocean-colour observations into oceanic ecosystem models. AB - The effectiveness of ocean-colour data assimilation in providing robust biological-parameter estimates for basin-scale ecosystem models is investigated for a phytoplankton-zooplankton-nutrient model using North Atlantic satellite chlorophyll data. The model is forced by annual cycles of mixed-layer depth, day length, photosynthetically available radiation and a temperature-dependent phytoplankton maximum growth rate. Although ocean-colour data are potentially limited in their ability to constrain model parameters because they provide information about the phytoplankton component only, this limitation is offset by the volume of data available covering the range of possible biogeochemical responses to similar and widely varying physical conditions. The results are improved by applying wintertime nutrient estimates based on in situ observations as an additional constraint. Repeatability of parameter estimates obtained from independent samples is examined. Results obtained using regional and basin-wide sampling strategies for obtaining the optimization dataset are compared and the geographic applicability of the calibrated models is assessed. PMID- 12626240 TI - Rossby waves: synergy in action. AB - Rossby waves are an important phenomenon, linking processes in the west of ocean basins with forcing that occurred earlier in the east. We show evidence for such features in satellite-derived datasets of sea-surface height, temperature and ocean colour, using a section of the south Indian Ocean as an example. We discuss the possible mechanisms for an effect on chlorophyll, and we investigate this by comparing the ocean colour signal with the other datasets. In this region, the primary mechanism for a Rossby-wave signal in ocean colour appears to be meridional advection of water across a strong chlorophyll gradient. However, this cannot fully explain the observations in the westernmost basin. PMID- 12626242 TI - Initial in-orbit results from a low-cost atmospheric ozone monitor operating on board the FASat-Bravo microsatellite. AB - In 1994, a collaborative programme was set up between the University of Surrey and the Chilean Air Force to design and build a low-cost 50 kg microsatellite with instrumentation capable of monitoring the distribution and concentration of stratospheric ozone, particularly over Chilean territory. This resulted in the joint design and development of the ozone-layer monitoring experiment (OLME), which was flown on board the resultant FASat-Bravo microsatellite, launched in July 1998 into an 820 km altitude Sun-synchronous orbit. The nadir-pointing OLME measures the solar ultraviolet light backscattered from the atmosphere in four spectral bands. From these raw radiometric data, total column ozone concentrations can be deduced. Since the launch, a series of high-spatial resolution measurements have been made over Chilean scientific stations, while continuous low-spatial-resolution measurements have been used to provide global coverage. To date, the University of Surrey has concentrated on these global measurements, and has used a simplified ozone retrieval algorithm to check the results qualitatively against ozone maps produced by NASA's Earth Probe Total Ozone Mapping Spectrometer (TOMS) payload. These initial results look promising, with apparently good correlation between the two datasets. A novel, more-complex retrieval algorithm has been applied to the high-resolution data, and early results have shown agreement with the TOMS. PMID- 12626251 TI - Earthquake science research with a microsatellite. AB - Reliable, repeatable earthquake forecast is a subject surrounded by controversy and scepticism. What is clear is that reliable forecast would be a critical tool for effective earthquake disaster management. It is proposed that satellites and ground-based facilities may detect earthquake precursors in the ionosphere a few hours or days before the main shock. A low-cost 100 kg class satellite carrying a topside sounder is proposed, to make systematic measurements over seismically active zones. The mission aims to confirm or refute the hypothesis of ionospheric earthquake precursors, define the reliability and reproducibility, and enable further scientific understanding of their mechanisms. PMID- 12626249 TI - A strategy for the observation of volcanism on Earth from space. AB - Heat, strain, topography and atmospheric emissions associated with volcanism are well observed by satellites orbiting the Earth. Gravity and electromagnetic transients from volcanoes may also prove to be measurable from space. The nature of eruptions means that the best strategy for measuring their dynamic properties remotely from space is to employ two modes with different spatial and temporal samplings: eruption mode and background mode. Such observational programmes are best carried out at local or regional volcano observatories by coupling them with numerical models of volcanic processes. Eventually, such models could become multi-process, operational forecast models that assimilate the remote and other observables to constrain their uncertainties. The threat posed by very large magnitude explosive eruptions is global and best addressed by a spaceborne observational programme with a global remit. PMID- 12626259 TI - A worthwhile investment: research-council scientists speak out. AB - This article looks at the work of two of the UK's research councils and the way their funding contributes to science at large, and to the scientists themselves. Nina Morgan talks to three scientists from the Particle Physics and Astronomy Research Council and three from the Natural Environment Research Council about their interests and enthusiasms. PMID- 12626260 TI - Cosmology with varying constants. AB - The idea of possible time or space variations of the 'fundamental' constants of nature, although not new, is only now beginning to be actively considered by large numbers of researchers in the particle physics, cosmology and astrophysics communities. This revival is mostly due to the claims of possible detection of such variations, in various different contexts and by several groups. I present the current theoretical motivations and expectations for such variations, review the current observational status and discuss the impact of a possible confirmation of these results in our views of cosmology and physics as a whole. PMID- 12626261 TI - The dark side of galaxy formation. AB - I discuss the discovery of a population of extremely luminous, but very dusty and very distant, galaxies in the submillimetre (submm) waveband. Almost all the light emitted by the stars in these galaxies is absorbed by interstellar dust (which is produced by the same stars) and re-radiated in the far-infrared. This leaves little to be detected at optical wavelengths and results in most of these galaxies being effectively invisible in even the deepest optical images obtainable with the Hubble space telescope. Yet this population contributes most of the light emitted by galaxies at wavelengths of lambda > or approximately equal 100 microm over the lifetime of the Universe. Together with other observations, this suggests that perhaps up to half of all the stars seen in galaxies today were formed in very dusty regions in the early Universe. Hence, studying the galaxies detected in the submm wavebands is critical for developing and testing models of galaxy formation and evolution. Individually, these luminous submm galaxies are forming stars a thousand times faster than our Galaxy is at the present-day, sufficiently fast to form all the stars in the most luminous galaxy in the local Universe within a short period, up to ca. 0.1-1 Gyr. Detailed study of a handful of examples of this population confirm these estimates and unequivocally identify the bulk of this submm-selected population with dusty, star-burst galaxies in the very distant Universe. The extreme faintness of this population in the optical and near-infrared wavebands, resulting from their obscuration by dust, means that our understanding of the detailed nature of these galaxies is only slowly growing. I give a brief summary of the properties of these highly obscured systems and describe the wide range of facilities currently being developed that will greatly aid in their study. PMID- 12626262 TI - The resolved history of galaxy evolution. AB - We briefly review the study of the evolution of galaxies from an observational point of view, with particular emphasis on the role of the Hubble Space Telescope in probing the evolution of the different morphological types of galaxy. We show how using the stellar mass of galaxies as a tracer of evolution can improve our understanding of the physical process taking place before turning our eyes towards the future and giving an overview of what we can expect from future advances in technology. PMID- 12626264 TI - The solar dynamo. AB - In this article I review the fundamentals of solar-dynamo theory. I describe both historical and contemporary observations of the solar magnetic field before outlining why it is believed that the solar field is maintained by a hydromagnetic dynamo. Having explained the basic dynamo process and applications of the theory to the Sun, I shall conclude by speculating on future directions for the theory. PMID- 12626263 TI - Violence in the hearts of galaxies: aberration or adolescence? AB - Violent activity in the nuclei of galaxies has long been considered a curiosity in its own right; manifestations of this phenomenon include distant quasars in the early Universe and comparatively nearby Seyfert galaxies, both thought to be powered by the release of gravitational potential energy as material from the host galaxy accretes onto a central supermassive black hole (SMBH). Traditionally, the broader study of the formation, structure and evolution of galaxies has largely excluded active galactic nuclei. Recently, however, this situation has changed dramatically, both observationally and theoretically, with the realization that the growth and influence of the SMBH, the origin and development of galaxies and nuclear activity at different epochs in the Universe may be intimately related. The most spectacular fireworks seen in distant quasars may be relatively easy to explain, since the era of greatest quasar activity seems to coincide with turbulent dynamics at the epoch of galaxy formation in the young, gas-rich Universe. Ubiquitous black holes are believed to be a legacy of this violent birth. Alternatively, black holes may be the seeds that drive galaxy formation in the first place. Closer to home, and hence more recently in the history of the Universe, a fraction of comparatively ordinary galaxies, similar to our own, has reignited their central engines, albeit at a lower level of activity. Since these galaxies are more established than their younger and more distant counterparts, the activity here is all the more puzzling. Whatever the mechanisms involved, they are likely to play an important role in galaxy evolution. I review the intriguing evidence for causal links between SMBHs, nuclear activity and the formation and evolution of galaxies, and describe opportunities for testing these relationships using the next generation of earthbound and space-borne astronomical facilities. PMID- 12626265 TI - The astrophysics of crowded places. AB - Today the Sun is in a relatively uncrowded place. The distance between it and the nearest other star is relatively large (about 200,000 times the Earth-Sun distance!). This is beneficial to life on Earth; a close encounter with another star is extremely unlikely. Such encounters would either remove the Earth from its orbit around the Sun or leave it on an eccentric orbit similar to a comet's. But the Sun was not formed in isolation. It was born within a more-crowded cluster of perhaps a few hundred stars. As the surrounding gas evaporated away, the cluster itself evaporated too, dispersing its stars into the Galaxy. Virtually all stars in the Galaxy share this history, and here I will describe the role of 'clusterness' in a star's life. Stars are often formed in larger stellar clusters (known as open and globular clusters), some of which are still around today. I will focus on stars in globular clusters and describe how the interactions between stars in these clusters may explain the zoo of stellar exotica which have recently been observed with instruments such as the Hubble Space Telescope and the X-ray telescopes XMM-Newton and Chandra. In recent years, myriad planets orbiting stars other than the Sun--the so-called 'extrasolar' planets--have been discovered. I will describe how a crowded environment will affect such planetary systems and may in fact explain some of their mysterious properties. PMID- 12626266 TI - Solar variability, coupling between atmospheric layers and climate change. AB - One of the enduring puzzles of atmospheric physics is the extent to which changes in the Sun can influence the behaviour of the climate system. While solar-flux changes tend to be relatively modest, a number of observations of atmospheric parameters indicates a disproportionately large response. Global-scale models of the coupled middle and upper atmosphere have provided new insights into some of the mechanisms that may be responsible for the amplification of the solar signal. In particular, modification of the transport of heat and chemicals such as ozone by waves during periods of solar activity has been shown to make an important contribution to the climate of the stratosphere and mesosphere. In this paper, a review of some of the recent advances in understanding the coupling between atmospheric layers and how this work relates to Sun-weather relations and climate change in the troposphere will be presented, along with a discussion of some of the challenges that remain. PMID- 12626267 TI - Imaging of near-Earth space plasma. AB - This paper describes the technique of imaging the ionosphere using tomographic principles. It reports on current developments and speculates on the future of this research area. Recent developments in computing and ionospheric measurement, together with the sharing of data via the internet, now allow us to envisage a time when high-resolution, real-time images and 'movies' of the ionosphere will be possible for radio communications planning. There is great potential to use such images for improving our understanding of the physical processes controlling the behaviour of the ionosphere. While real-time images and movies of the electron concentration are now almost possible, forecasting of ionospheric morphology is still in its early stages. It has become clear that the ionosphere cannot be considered as a system in isolation, and consequently new research projects to link together models of the solar-terrestrial system, including the Sun, solar wind, magnetosphere, ionosphere and thermosphere, are now being proposed. The prospect is now on the horizon of assimilating data from the entire solar-terrestrial system to produce a real-time computer model and 'space weather' forecast. The role of tomography in imaging beyond the ionosphere to include the whole near-Earth space-plasma realm is yet to be realized, and provides a challenging prospect for the future. Finally, exciting possibilities exist in applying such methods to image the atmospheres and ionospheres of other planets. PMID- 12626268 TI - Mixing in vortical, chaotic and turbulent flows. AB - Mixing is discussed in relation to stirring as reflected in the geometry of advected interfaces, the behaviour of fluid-element pairs and their separation rates. Stirring is different in vortical, chaotic and turbulent flows because of qualitative differences in spatio-temporal flow structure, thus giving rise to different mixing laws. Important applications of the mixing and stirring properties discussed in this review are chlorine deactivation and ozone depletion in stratospheric mid-latitudes. PMID- 12626269 TI - The break-up of continents and the formation of new ocean basins. AB - Rifted continental margins are the product of stretching, thinning and ultimate break-up of a continental plate into smaller fragments, and the rocks lying beneath them store a record of this rifting process. Earth scientists can read this record by careful sampling and with remote geophysical techniques. These experimental studies have been complemented by theoretical analyses of continental extension and associated magmatism. Some rifted margins show evidence for extensive volcanic activity and uplift during rifting; at these margins, the record of the final stages of rifting is removed by erosion and obscured by the thick volcanic cover. Other margins were underwater throughout their formation and showed rather little volcanic activity; here the ongoing deposition of sediment provides a clearer record. During the last decade, vast areas of exhumed mantle rocks have been discovered at such margins between continental and oceanic crust. This observation conflicts with the well-established idea that the mantle melts to produce new crust when it is brought close to the Earth's surface. In contrast to the steeply dipping faults commonly seen in zones of extension within continental interiors, faults with very shallow dips play a key role in the deformation immediately preceding continental break-up. Future progress in the study of continental break-up will depend on studies of pairs of margins which were once joined and on the development of computer models which can handle rigorously the complex transition from distributed continental deformation to sea floor spreading focused at a mid-ocean ridge. PMID- 12626270 TI - On the time-scales of magmatism at island-arc volcanoes. AB - Precise information on time-scales and rates of change is fundamental to an understanding of natural processes and the development of quantitative physical models in the Earth sciences. U-series isotope studies are revolutionizing this field by providing time information in the range 10(2)-10(4) years, which is similar to that of many modern Earth processes. I review how the application of U series isotopes has been used to constrain the time-scales of magma formation, ascent and storage beneath island-arc volcanoes. Different elements are distilled off the subducting plate at different times and in different places. Contributions from subducted sediments to island-arc lava sources appear to occur some 350 kyr to 4 Myr prior to eruption. Fluid release from the subducting oceanic crust into the mantle wedge may be a multi-stage process and occurs over a period ranging from a few hundred kyr to less than one kyr prior to eruption. This implies that dehydration commences prior to the initiation of partial melting within the mantle wedge, which is consistent with recent evidence that the onset of melting is controlled by an isotherm and thus the thermal structure within the wedge. U-Pa disequilibria appear to require a component of decompression melting, possibly due to the development of gravitational instabilities. The preservation of large (226)Ra disequilibria permits only a short period of time between fluid addition and eruption. This requires rapid melt segregation, magma ascent by channelled flow and minimal residence time within the lithosphere. The evolution from basalt to basaltic andesite probably occurs rapidly during ascent or in magma reservoirs inferred from some geophysical data to lie within the lithospheric mantle. The flux across the Moho is broadly andesitic, and some magmas subsequently stall in more shallow crustal level magma chambers, where they evolve to more differentiated compositions on time-scales of a few thousand years or less. PMID- 12626271 TI - Remote monitoring of the earthquake cycle using satellite radar interferometry. AB - The earthquake cycle is poorly understood. Earthquakes continue to occur on previously unrecognized faults. Earthquake prediction seems impossible. These remain the facts despite nearly 100 years of intensive study since the earthquake cycle was first conceptualized. Using data acquired from satellites in orbit 800 km above the Earth, a new technique, radar interferometry (InSAR), has the potential to solve these problems. For the first time, detailed maps of the warping of the Earth's surface during the earthquake cycle can be obtained with a spatial resolution of a few tens of metres and a precision of a few millimetres. InSAR does not need equipment on the ground or expensive field campaigns, so it can gather crucial data on earthquakes and the seismic cycle from some of the remotest areas of the planet. In this article, I review some of the remarkable observations of the earthquake cycle already made using radar interferometry and speculate on breakthroughs that are tantalizingly close. PMID- 12626272 TI - Anthropogenic lead dynamics in the terrestrial and marine environment. AB - Human activities have greatly altered the natural geochemical cycles of several heavy metals, most notably lead derived from leaded-petrol and metal-smelting emissions. This inadvertent geochemical tracer experiment poses two challenges: understanding how anthropogenic lead affects human health and the environment, and quantifying its time-dependent distribution within terrestrial and marine systems. Accurate assessment of the latter relies on well-constrained historical and modern lead fluxes from proxy records and direct observations, lead source estimates from stable lead isotopes, and transport rate estimates from radionuclides. Numerous studies support the global-scale atmospheric lead fluxes principally derived from anthropogenic activities, the short lead residence time in the atmosphere and surface ocean, and the predominance of North American and European lead emissions. Emerging observations and models are currently addressing the time-dependent evolution of this reactive tracer in the atmosphere and oceans. PMID- 12626273 TI - Dust in the Earth system: the biogeochemical linking of land, air and sea. AB - Understanding the response of the Earth's climate system to anthropogenic perturbation has been a pressing priority for society since the late 1980s. However, recent years have seen a major paradigm shift in how such an understanding can be reached. Climate change demands analysis within an integrated 'Earth-system' framework, taken to encompass the suite of interacting physical, chemical, biological and human processes that, in transporting and transforming materials and energy, jointly determine the conditions for life on the whole planet. This is a highly complex system, characterized by multiple nonlinear responses and thresholds, with linkages often between apparently disparate components. The interconnected nature of the Earth system is wonderfully illustrated by the diverse roles played by atmospheric transport of mineral 'dust', particularly in its capacity as a key pathway for the delivery of nutrients essential to plant growth, not only on land, but perhaps more importantly, in the ocean. Dust therefore biogeochemically links land, air and sea. This paper reviews the biogeochemical role of mineral dust in the Earth system and its interaction with climate, and, in particular, the potential importance of both past and possible future changes in aeolian delivery of the micro-nutrient iron to the ocean. For instance, if, in the future, there was to be a widespread stabilization of soils for the purpose of carbon sequestration on land, a reduction in aeolian iron supply to the open ocean would occur. The resultant weakening of the oceanic carbon sink could potentially offset much of the carbon sequestered on land. In contrast, during glacial times, enhanced dust supply to the ocean could have 'fertilized' the biota and driven atmospheric CO(2) lower. Dust might even play an active role in driving climatic change; since changes in dust supply may affect climate, and changes in climate, in turn, influence dust, a 'feedback loop' is formed. Possible feedback mechanisms are identified, recognition of whose operation could be crucial to our understanding of major climatic transitions over the past few million years. PMID- 12626274 TI - Carbon in the atmosphere and terrestrial biosphere in the 21st century. AB - The release of carbon dioxide from fossil-fuel combustion and land-use change has caused a significant perturbation in the natural cycling of carbon between land, atmosphere and oceans. Understanding and managing the effects of this disruption on atmospheric composition and global climate are likely to be amongst the most pressing issues of the 21st century. However, the present-day carbon cycle is still poorly understood. One remarkable feature is that an increasing amount of atmospheric carbon dioxide appears to be being absorbed by terrestrial vegetation. I review the recent evidence for the magnitude and spatial distribution of this 'terrestrial carbon sink', drawing on current research on the global atmospheric distribution and transport of carbon dioxide, oxygen and their isotopes; direct measurement of CO(2) fluxes above various biomes; and inventories of forest biomass and composition. I review the likely causes of these carbon sinks and sources and their implications for the ecology and stability of these biomes. Finally, I examine prospects and key issues over coming decades. Within a few years, satellite measurements of atmospheric CO(2) and forest biomass, coupled with 'real-time' biosphere-atmosphere models, will revolutionize our understanding of the terrestrial carbon cycle. Controlling deforestation and managing forests has the potential to play a significant but limited part in reaching the goal of stabilizing atmospheric CO(2) concentrations. However, there are likely to be limits to the amount of carbon storage possible in natural vegetation, and, in the long term, terrestrial carbon storage may be unstable, with the potential to accelerate rather than brake global warming. PMID- 12626275 TI - Costing climate change. AB - Debate over how, when, and even whether man-made greenhouse-gas emissions should be controlled has grown in intensity even faster than the levels of greenhouse gas in our atmosphere. Many argue that the costs involved in reducing emissions outweigh the potential economic damage of human-induced climate change. Here, existing cost-benefit analyses of greenhouse-gas reduction policies are examined, with a view to establishing whether any such global reductions are currently worthwhile. Potential for, and cost of, cutting our own individual greenhouse-gas emissions is then assessed. I find that many abatement strategies are able to deliver significant emission reductions at little or no net cost. Additionally, I find that there is huge potential for individuals to simultaneously cut their own greenhouse-gas emissions and save money. I conclude that cuts in global greenhouse-gas emissions, such as those of the Kyoto Protocol, cannot be justifiably dismissed as posing too large an economic burden. PMID- 12626276 TI - Earth's biggest 'whodunnit': unravelling the clues in the case of the end-Permian mass extinction. AB - The mass extinction that occurred at the end of the Permian period, 250 million years ago, was the most devastating loss of life that Earth has ever experienced. It is estimated that ca. 96% of marine species were wiped out and land plants, reptiles, amphibians and insects also suffered. The causes of this catastrophic event are currently a topic of intense debate. The geological record points to significant environmental disturbances, for example, global warming and stagnation of ocean water. A key issue is whether the Earth's feedback mechanisms can become unstable on their own, or whether some forcing is required to precipitate a catastrophe of this magnitude. A prime suspect for pushing Earth's systems into a critical condition is massive end-Permian Siberian volcanism, which would have pumped large quantities of carbon dioxide and toxic gases into the atmosphere. Recently, it has been postulated that Earth was also the victim of a bolide impact at this time. If further research substantiates this claim, it raises some intriguing questions. The Cretaceous-Tertiary mass extinction, 65 million years ago, was contemporaneous with both an impact and massive volcanism. Are both types of calamity necessary to drive Earth to the brink of faunal cataclysm? We do not presently have enough pieces of the jigsaw to solve the mystery of the end-Permian extinction, but the forensic work continues. PMID- 12626277 TI - Longevity and clinical performance of IPS-Empress ceramic restorations--a literature review. AB - A literature review of longevity and clinical performance of IPS-Empress restorations is presented. A MEDLINE search was conducted in fall 2000. Selection criteria were set so as to identify suitable clinical trials that were published in full and that had lasted more than 2 years. A total of 6 clinical trials on the performance of IPS-Empress inlays and onlays and a total of 3 clinical trials on the performance of IPS-Empress crowns were identified. Survival rates for IPS Empress inlays and onlays ranged from 96% at 4.5 years to 91% at 7 years; most failures were due to bulk fracture. IPS-Empress crowns had a survival rate ranging from 92% to 99% at 3 to 3.5 years; crown failure was also mainly due to fracture. Dentists should inform their patients about these survival rates when offering such treatment. The use of IPS-Empress crowns in the posterior of the mouth is not recommended until the results of more long-term clinical trials are available. PMID- 12626278 TI - Use of photographs for communicating with the laboratory in indirect posterior restorations. AB - This article presents a single onlay case that was significantly enhanced through a detailed communication process between the clinician and the laboratory technician. By using colour slides as part of the dentist-technician communication process, the author found that the technician was better able to create an esthetic, accurate and successful restoration that addressed the patient's needs. PMID- 12626279 TI - Ameloblastic fibro-odontoma: a case report. AB - Ameloblastic fibro-odontoma is a benign epithelial odontogenic tumour with odontogenic mesenchyme exhibiting the histologic characteristics of ameloblastic fibroma and complex odontoma. It is usually associated with developing teeth and occurs predominantly in children and adolescents. In many cases, such lesions are found on radiographic evaluation of patients in whom eruption of teeth is delayed. Ameloblastic fibro-odontoma is generally asymptomatic but may cause swelling and discomfort. This report describes an ameloblastic fibro-odontoma in the posterior mandible of a 26-year-old woman and discusses the histogenesis and clinical features of the lesion. PMID- 12626280 TI - Acute herpetic gingivostomatitis in adults: a review of 13 cases, including diagnosis and management. AB - OBJECTIVE: To present to general dentists the typical signs and symptoms associated with adult acute (primary) herpetic gingivostomatitis. The pertinent laboratory tests, management options and current pharmacotherapy are also reviewed. REVIEW DESIGN: The clinical files of 13 adult patients were reviewed. All had no history of herpes simplex virus infection and presented with oral lesions suggestive of primary herpetic infection. The subjects were all patients of one of the investigators, and their workup included Tzanck testing and viral culture. RESULTS: The patients ranged in age from 18 to 79 (mean 37.2, standard deviation 19.6) years. Nine (69%) were men. Viral culture was confirmed as the gold standard for diagnosis. The sensitivity of Tzanck testing was 77% (10/13), slightly higher than that reported previously (40% to 50%). In this patient group the febrile lymphadenopathic profile was typical of younger patients (18 to 42 years of age), whereas older patients presented with predominantly oral symptoms. CONCLUSIONS: Primary herpetic gingivostomatitis is not limited to children but can affect people of any age. Proper diagnosis and treatment are essential, particularly in elderly and immunocompromised patients. Tzanck testing may serve as a useful adjunct in diagnosis. Antiviral agents such as valacyclovir and famciclovir should be considered part of early management. Dentists are often the first health care professionals to be consulted by patients with this condition, and recognition of the infection is paramount. PMID- 12626281 TI - Amprenavir in pre-treated patients: virological and immunological response in a cohort of 45 patients. AB - Treatment failure after preceding protease inhibitors (PI) is often due to resistance mutations. Our objective was to evaluate amprenavir (APV) in pre treated patients and to correlate it with pre-existing mutations. - Fourty five patients were entered in an open label prospective study (6/99-12/2000). Pre treatment was 6.2 years +/- 2.4 (2-11.3) and included a mean of 4.13 nucleosides (RTIs) and 2.73 PIs. Genotypic resistance testing was performed prior to the switch. APV dose was 1200 mg/d in combination with ritonavir (RTV) boosting (2 x 100mg) and 2 x 1200 mg/d in 6 patients without RTV. Co-medication was selected based on treatment history and results of genotypic testing. - The median duration of treatment at analysis was 34 weeks. Plasma viral load (VL) average at baseline was 4.6 log subset 10 +/- 08. After 24 weeks the mean VL reduction was 1.4 log subset 10 +/- 1.2 3.86-0.40s). A VL reduction of >1.5 log subset 10 was found in 16/45 patients (36%), 21/45 (47%) patients achieved a VL <400 cp/ml, and 12/45 (27%) a VL < 50 cp/ml. CD4 cells increased from a mean baseline of 208/microl +/- 185 to 318/microl +/- 253. Fourty percent (18/45) of patients had a CD4 gain of more than 100 cells/microl. Genotypic resistance determination showed PI mutations in 87% of patients tested. The average number of mutated codons was 4.54. Three of 4 patients with I84V mutation did not achieve an undetectable VL. - Our findings demonstrate that APV and APV/RTV plus two additional antiretrovirals has a good virological and immunological success rate in pre-treated patients. Presence of more than two APV resistance mutations was associated with treatment failure. PMID- 12626282 TI - Superior virological efficacy of ritonavir-boosted protease inhibitor regimens compared to single protease inhibitor therapy. AB - Low dose-ritonavir boosted protease inhibitors are increasingly being used for the first-line antiretroviral treatment, though their virological efficacy has just poorly been compared to alternative antiretroviral therapies. Here, we retrospectively investigated the virological responses of 316 protease inhibitor naive HIV patients receiving highly active antiretroviral treatment based on a single (n = 256) or a ritonavir-boosted protease inhibitor (n = 60), both in the background of two nucleoside analogues. - By intent-to-treat analysis, a complete initial virological response was achieved in 71.8% of all patients in the single protease inhibitor group (indinavir: 76%, ritonavir: 67.5%, nelfinavir: 70.6%) and in 88.3% (p = 0.008) of patients treated with a boosted protease inhibitor (saquinavir/r: 71.4%, indinavir/r: 92.1%, lopinavir/r: 86.6%). The multivariate risk analysis identified boosted PI treatment as an independent predictor of a complete virological response (OR = 2.8, p=0.02). Viral rebound after an initial complete virological response was observed in 28% and 17% (p = 0.06) of patients receiving a single or a dual protease inhibitor, respectively. The rate of durable viral suppression over 12 months was 44.5% and 56.7% (p = 0.09) in the respective study cohorts. Ritonavir-boosted protease inhibitors therefore seem to induce a superior virological response rate and a higher degree of sustained virological suppression. PMID- 12626283 TI - Clinical correlation of antimitochondrial antibodies. AB - BACKGROUND: Antimitochondrial antibodies (AMA) are a hallmark of primary biliary cirrhosis (PBC). They are believed to be absolutely disease specific. It does occur that patients with positive AMA are diagnosed with PBC in the absence of liver specific signs and symptoms. The aim of the present study was to examine the disease spectrum of unselected AMA positive patients of an university hospital. METHODS: All of the AMA tests performed in the immunological laboratory of the hospital between 1992 and 1998 were examined for positivity. 100 patients with a positive result were analyzed retrospectively for diagnosis, clinical and laboratory features. RESULTS: 61 patients suffered from liver diseases and 39 from non-liver diseases. The patients with liver diseases were 36 patients with PBC, 2 patients with PBC/PSC-overlap syndrome, 4 patients with autoimmune hepatitis and 19 patients with different liver diseases of other than autoimmune origin. The 39 patients with non-liver diseases included 9 patients with systemic autoimmune diseases, 3 patients with organ-specific autoimmune diseases, 8 patients with carcinoma and 19 patients with different diseases. 97 patients had an ELISA test for antibodies to the mitochondrial antigen M2 performed in addition to the immunofluorescence test for AMA. 73 patients had positive values for anti-M2 antibodies and 24 patients had negative results. Anti-M2 antibody values were divided in negative, low (5-100 U/ml), medium (101-1000 U/ml), high (1001-10000 U/ml) and very high (>10000 U/ml). Very high and high anti-M2 values were present mainly in patients with PBC and some patients with other liver diseases, medium high and low values in patients with different disease groups. In this unselected patient population only one third of AMA positive patients had an established diagnosis of PBC, about 10% a diagnosis of a systemic autoimmune disease and 3 % had other organ-specific autoimmune diseases. It can be concluded that, although high titers of antibodies against M2 are present mainly in patients with PBC, medium high and low titers may be present in patients lacking symptoms of PBC. CONCLUSIONS: Although there is the possibility that the patients will develop the disease in the future, the presence of AMA alone does not seem to be sufficient to establish a diagnosis of PBC in a patient population in which at a given time point only 38 % of AMA positive patients have the disease. PMID- 12626284 TI - Different expression of cytokines in survivors and non-survivors from MODS following cardiovascular surgery. AB - OBJECTIVE: Cardiopulmonary bypass is often associated with pathophysiological changes in form of systemic inflammatory response syndrome (SIRS) or multiple organ dysfunction syndrome (MODS). In the present study, we investigated plasma levels of pro- and anti-inflammatory cytokines in survivors and non-survivors from MODS in the early postoperative course following open heart surgery. DESIGN: Prospective clinical study. SETTING: A University Cardiothoracic Intensive Care Unit. METHODS: Levels of cytokines (IL-6, IL-8, IL-10, IL-18, and TGF- ) and procalcitonin (PCT) were measured at the first four postoperative days in 16 adult male patients with an Apache II-score >24 and two or more organ dysfunctions after myocardial revascularization. MAIN RESULTS: All pro inflammatory cytokines, except for IL-6, were significantly elevated in non survivors from MODS, with peak values at the first two postoperative days. The plasma levels of immunoinhibitory cytokines showed no differences between the groups. CONCLUSIONS: The results of our study show a different expression of pro inflammatory cytokines in survivors and non-survivors from MODS following operations with extracorporeal circulation. In addition to Apache-II score, especially IL-8, IL-18, and PCT may be used as parameters for the prognosis of patients with organ dysfunctions after cardiac surgery. PMID- 12626285 TI - Longitudinal evaluation of serum estradiol and estrone in male patients infected with the human immunodeficiency virus. AB - In patients with human immunodeficiency virus (HIV) infection alteration of various endocrine functions have been described. However, there is limited information available on estrogens and their function in these patients. The aim of this study was to evaluate the pituitary and testicular endocrine markers: 14 HIV-positive men were included into the longitudinal study with a follow up to of 18 month period. None of the patients had a history or clinical evidence of endocrine dysfunction. Follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin, estrone, estradiol, and testosterone were measured in serum by commercially available radioimmunoassays. Prolactin levels were not affected in the patients. LH and FSH remained within normal range, but at the end of the study period we observed a significant increase of LH (p < 0.05). In addition, testosterone levels were normal with a slight reduction at the end of the 18 month observation period (p <0.05). Both estrone and estradiol were significantly increased with a further rise at the end of the study (p <0.05). In summary, the elevation of the serum estrone and estradiol in HIV-afflicted patients correlated with the progress of the HIV-disease, being paralleled by a decrease in testosterone. The altered relation of estrogens and testosterone may in a part be responsible for the complaints of decreased libido and increased impotence often observed in HIV-infected men. PMID- 12626286 TI - Salivary peroxidase levels in patients with AIDS. AB - The main consequences of human immunodeficiency virus (HIV) infection and AIDS are frequent and persistent opportunistic infections at mucosal surfaces, but data upon impaired oral mucosal response in AIDS patients are still lacking. - The aim of this study was to determine salivary flow rates and peroxidase levels in unstimulated whole saliva in AIDS patients together with comparison to the healthy controls. Salivary peroxidase levels were determined according to Putter and Becker in 20 AIDS patients and 18 HIV-seronegative healthy controls. Statistical analysis was performed using Student t-test. Salivary peroxidase levels were significantly increased in the AIDS group (9.41 +/- 8.50 kU/L; p<0.009) when compared to the healthy controls (3.1 +/- 2.0 kU/L). Salivary flow rates were significantly decreased in AIDS patients (0.17+/-0.11 ml/min, p<0.009) when compared with healthy controls (0.58 +/- 0.19 ml/min). Elevated salivary peroxidase levels indicate increased salivary antimicrobial activity in AIDS patients. PMID- 12626287 TI - Review of existing prophylaxis programs for 3880 first graders from Rhineland Palatinate, Germany. AB - A large study amongst 6-7 years old children was conducted in one of the German Federal States to evaluate the success of presently existing prophylaxis programs and to determine groups at high risk for dental caries. - A total of 3880 children (1844 girls and 2036 boys) received a dental exam while in first grade in the year 2000. 37.8% (n = 1465) of the students lived in a larger city (approximately 200000 inhabitants), 62.2% (n = 2415) of the students came from rural areas. The proportion of foreign children, mainly from Serbia, Croatia and Turkey, amounted to a total of 19.1% (n = 741). All children were cared for as part of a group dental prophylaxis program in their respective schools and were examined under the protocol of the dmf-t/DMF-T index. 53.4% (n = 2073) of the students had a naturally healthy dentition, with the percentage of boys (55.5%, n = 1129) being higher than that of girls (51.2%, n = 944). The percentage of the caries-free dentitions in the rural population was 56.1% (n = 1163) whereas the percentage amongst the urban students was 43.9% (n = 910). The percentage of naturally healthy dentitions was 60.8% (n = 1909) amongst German children, and their foreign classmates had 21.9% (n = 162) caries-free dentitions. 37.6% (n = 1459) of all children had carious lesions; the foreign children had carious defects in 69.7% (n = 517) of the cases and the German students in 30.1% (n = 944). 8.8% of all first graders surveyed had fully treated healthy dentitions. - Although the goal of the WHO to obtain caries-free dentitions amongst 5-6 year olds in 50% of the cases has been met in the year 2000, this study showed clearly that oral health was significantly worse for the majority of foreign children compared to the German school children. PMID- 12626288 TI - [Traditional enema for newborns and infants in Bobo Dioulasso: health practice or socialisation]. AB - Kanki demonstrated a high prevalence and frequency of enema practised with newborns in the South-West of Burkina Faso. Little is known about the risks on children's health possibly associated with this practice and about its impact on other treatments in paediatrics. In this study, the authors describe daily administered enema (DAE) and analyse local conceptual frameworks underlying this practice through in-depth-interviews and focus group discussions with 30 mothers, 5 traditional healers and 5 health agents. Various medications are used to compose the liquid introduced by the mothers in the child's anus. Many of these substances are prone to irritate intestinal mucus, others are simply toxic. Practically, enema aims at curing or preventing a variety of diseases caused by an accumulation of impurities (nogo) in the intestines due to the consumption of inappropriate food. With newborns, diseases are transmitted by mothers through breastfeeding after eating food which is too sweet or too fat. In addition to provoking diseases, the nogo also "block" the child's physical and psychic development during his/her first year of life. Therefore, as soon as the child has excreted for the first time, most mothers give enema daily both to protect their children from diseases and to speed their development. In fact, beside prophylaxy and therapy lies a "didactic" function of enema as a medication used to help the child to stand up, to get teeth..., to gain independence from his/her mother. DAE therefore plays an important role in the process of acquiring bio social aptitudes, i.e., important educational virtues to achieve a successful first step in the socialisation process. Exploring more deeply local perceptions explaining the origin of the n g , the authors found an interesting relationship with religious taboos. Beside prohibited food, the n g are also due to transgression of various taboos surrounding birth and breastfeeding and even suggest a religious, rather than hygienic, explanation for the food prohibited. DAE ultimately consists in re-adjusting the child and his mother according to moral and cultural rules, avoiding the negative consequences of transgression. Consistently, DAE also facilitates the process of acquiring bio-social aptitudes for the child and therefore help him to enter his/her family as a fully accepted member. Moreover, religious prohibitions surrounding birth end as soon as the child is able to eat solid food and free him/herself. Therefore, DAE also helps the parents to get back to their normal life conditions. In conclusion, in addition to prophylactic and therapeutic explanations, the DAE participates in a general process of socialisation of newborns. To a certain extent, the child's health depends on the respect of the social and religious system and rules. The study reveals the important conceptual gap that may exist between two different logical frameworks--biomedical and popular--prone to explain health risk for newborns. The authors then reflect on the possible impact of health education programmes seeking to intervene at the hygienic level and ultimately facing an important set of cultural values aiming at keeping the social and cultural organisation coherent. PMID- 12626289 TI - [Monitoring the chemoresistance of Plasmodium falciparum malaria in Yopougon (Abidjan): in vivo study of chloroquine sensitivity and evaluation of pyrimethamine resistance following the analysis of point mutation in the dihydrofolate reductase gene]. AB - A major endemic in Cote d'Ivoire, malaria is the first cause of hospital admissions and mortality in tropical Africa. The decrease of morbidity and mortality depends on early diagnosis and relevant treatment. This situation is hampered by an emerging resistance of P. falciparum to usual drugs such as chloroquine and sulfadoxine-pyrimethamine. In recognition of this problem, we established a monitoring system in the north of Abidjan (Yopougon) in order to better analyse P. falciparum resistance. The molecular basis of P. falciparum resistance to pyrimethamine is associated with point mutations in the dihydrofolate reductase (dhfr) gene. The presence of a wild-type codon 108-ser is defined by the presence of an Alu1 restriction site. A single base change resulting in the change of amino acid from 108-Ser to 108-Asn or 108-Thr results in the appearance of a Bsr1 or a Scrf1 restriction site respectively. In response to these needs, 42 children aged 6 to 59 months were enrolled in the study by using tests of therapeutic efficacy of chloroquine (14-day in vivo test of WHO). Before treatment, infected blood samples were stored at 20 C until P. falciparum DNA extraction. The results of the in vivo sensitivity of chloroquine showed 84.3% of plasmodic rate, 97.7 % of P. falciparum against 2.3% of P. malariae. However, 78.6% of adequate clinical response (ACR) was obtained and 21.4% of early therapeutic failure (ETF). At the end of the study, clearance of parasitemia and fever was obtained but the gametocytic rate was 4.8%. More, RFLP studies of amplified DNA fragment revealed that P. falciparum from 12 children (44.5%) had point mutation in the codon 108 of the dhfr gene. The mutation of these isolates was based on the change of amino-acid from 108-Ser to 108-Asn. Moreover, 51.8% of isolates were of the wild type. In conclusion, our results showed that chloroquine resistance is a reality in Abidjan just like anywhere else in West Africa. However, the number of isolates which had point mutation in the dhfr gene suggested that the future approach must be the study of possible correlation between the in vivo sulfadoxine-pyrimethamine test and point mutations in the dihydrofolate reductase and in the dihydropteroate synthase gene of P. falciparum from Cote d'Ivoire. PMID- 12626290 TI - [Microbial quality assessment of some street food widely consumed in Ouagadougou, Burkina Faso]. AB - The major part of the population of Ouagadougou, Burkina Faso, have their breakfast, dinner and lunch in street food shops. The characteristics of street food vendors are indicated. It appeared clearly that women play a major part (75%) in street food sale. Vendors have only a slight knowledge of food processing and of hygienic practices. There is a high proportion (about 50%) of people among them. This dietary habit has some negative aspects on the consumers' health. Ours studies clearly showed that hygienic practices during sales operations are not respected by all categories of vendors of food products. Also, it was observed that street food vendors sometimes sit close to the waste water drainage system and solid wastes. Sometimes, the food is not covered and it is manipulated by consumers in areas infested by flies and others insects. The water used to wash the materials is of poor quality. The second aspects of our studies showed that, in most cases, when the safety and hygienic practices were not respected, the food became a true microorganism culture medium under tropical climatic conditions. A table reports microbial analysis and contamination rates of some important street foods consumed in Ouagadougou. In general these results showed the failure of microbial quality of some food which is not preheated (milk product, fruit juice, vegetable, fruit) and in the case of food which is not reheated after a long time of exposition (dry meat and meat on sticks). The presence of Salmonella and Shigella species in some food represents a serious danger for consumers. These aspects were observed with most street food vendors. It probably makes street food the source of most diseases caused by bacteria and other microorganisms. PMID- 12626291 TI - [Intensive care in Africa: a report of the first two years of activity of the intensive care unit of Ouagadougou national hospital (Burkina Faso)]. AB - OBJECTIVE: Intensive care units (ICUs) are very expensive and their role and effectiveness in developing countries are discussed; yet, their performance in these countries was infrequently reported. We report the experience over the first two years of activity of the multidisciplinary intensive care unit of the Ouagadougou national hospital. The analysis of such experience raises the issues related to intensive care in a developing country in terms of technical and social efficiency. DESIGN: Retrospective study of medical records. SETTING: Multidisciplinary ICUs of a national teaching hospital. The eleven million inhabitants of Burkina Faso are one the poorest nations in the world (3rd in UNDP classification). The Yalgado Ouedraogo national hospital is the largest in the country and the only one in the capital city, Ouagadougou; this national referral and teaching hospital has 724 beds. The ICU was created in December 1996; it has 8 beds, equipped with ventilators, monitors and various instruments. The staff consists of two full-time anesthesiologists and three others who contribute to the duty system, one senior nurse, two nurses specialized in anesthesia and fourteen other nurses. The unit is open to medical students and student nurses for hospital-based training. PATIENTS: All patients admitted in 1997 and 1998. MEASUREMENTS AND MAIN RESULTS: Data was collected from medical records and related to length of stay (LOS), morbidity, mortality, therapy and patients' socio-demographic background. No severity score was given. Three hundred and thirty-eight patients, mainly males (73%), were admitted; the average bed occupancy rate was 25%. The average age of patients was 39.05 +/- 1.21; there was no sex-specific age difference. Distribution as per socio-professional category showed a high proportion of civil servants (38.0%); farmers (23.7%) and housewives (17.6%) were relatively few. Admission diagnoses included 146 traumas (43.2%) of which 105 cranial traumas, 121 post operative (35.8%) and 71 medical pathologies (21.0%). Forty-nine patients (14.5%) were mechanically ventilated. The average LOS was 4.69 0.42 days; half of the patients stayed under 48 hours. The overall mortality rate was 63.6%. The rate was 79% for medical pathologies, 70.5% for traumas and 48.5% for post operative patients. The LOS was significantly longer in survivors (7.24 +/- 1.02) than in deceased (3.54 +/- 0.38). The cumulative survival curve showed a high death probability density which decreased in time: 0.74 on the first day, 0.58 on the second, 0.36 on the sixth. The mortality rate was higher in ventilated patients than in non ventilated ones. The highest mortality rate was observed among senior civil servants and farmers, and the lowest among craftsmen. The analysis of the first two years of operation of the ICUs of Ouagadougou national hospital reveals a low bed occupancy and a high mortality rate, particularly in the first days and for farmers. These results provide an opportunity to suggest the need for reorganization, with special emphasis on personnel availability and training, and for improved affordability of intensive care services. PMID- 12626292 TI - [Assessment and prospects of the community's participation at the Ziguinchor regional hospital centre (Senegal)]. AB - At the Ziguinchor regional hospital centre (RHC), the populations participate in the health effort by paying care services and medicines. The financial assessment of the participation of the community at the Ziguinchor RHC, from January 1,1998 to June 30, 2000, did not give good results. CFAF 263,066,092 of receipts have been generated, especially through the sale of medicines (38.85%) and via the obstetrical and gynecological clinic services (13.30%). The expenditures went up to CFAF 267,120,718; 41.87% were given to the staff, and 25.39% were used to buy medicines. Globally, expenditures exceeded receipts. At the beginning of January 1998, CFAF 7,600,524 were left to the Health Committee's budget. At the end of June 2000, there only remained CFAF 3,365,898. The decrease in the budget was expected because the RHC Health Committee board, set since 1984, never been renewed. Its management never followed the present recommendations. The board members, who were arbitrarily elected, didn't respect their commitment and were just represented by the treasurer. The latter had only a financial executive role. The board's decisions, which should have had the agreement of its president, were taken unilaterally by the direction of the RHC. Consequently, expenses evolved in keeping with gains, completely ignoring the use of the state budget. For a successful start of the RHC in the hospital reform, ten recommendations have been put across. The objective is to improve the organization of the Health Committee in order to better answer the population health problems. PMID- 12626293 TI - [Case-control study of hepatitis C HBs Ag and antivirus antibodies in chronic hepatopathies]. AB - The aim of this study is to establish the prevalence of hepatitis C HBs Ag and of anti-virus antibodies in chronic hepatopathies. The prospective case-control study was carried out on 91 patients who needed to be treated for chronic hepatopathies and 92 occasional blood donors. The search for hepatitis C HBs Ag and anti-virus antibodies was done using third generation ELISA screening. At the end of the study, HBs Ag was found in 54% of the patients vs. 4.3% of the control (p=0.0006). The two markers were present more frequently in cirrhosis than in hepatocellular carcinoma (HCC) and their association was more frequent in the case of cirrhosis. In Mali, hepatitis B and C viruses play an important part in chronic hepatopathies. PMID- 12626294 TI - [General practitioners' needs for continuing medical education in the Sousse region (Tunisia)]. AB - A continuing medical education is an essential activity in the search for doctors' performance, provided it is adapted to the specificity of their medical practice. The objective of this work is to identify the needs of general practitioners in relation to continuing medical education. It is a structural descriptive and transversal survey of about 112 general practitioners among the 140 doctors in the Sousse region in 2000. Data have been collected through a questionnaire that develops the expectation of general practitioners concerning the themes, domains and specialties privileged in a continued training. This research shows that the diagnostic and therapeutic strategies of current affections were themes requested by 85% of general practitioners. The doctor patient relation has been chosen by 71% of doctors. Emergency medicine was the specialty proposed by most of them. Thus, general practitioners expect a permanent training specific to their profile that could improve their clinical and relational competences. PMID- 12626295 TI - [Antenatal care utilization and unfavourable pregnancy outcome trends in Benin (Africa)]. AB - Antenatal care was an efficient preventive strategy for decreasing unfavorable pregnancy outcome incidence. Its appropriate utilisation remained the priority of Benin Health authorities following the Bamako Initiatives; but little research was undertaken on its effectiveness. This study proposes to describe the trends of unfavorable pregnancy outcome and antenatal care utilization. To that end, we collected and analysed retrospectively the data stored from 1993 to 1997 in the Heath Ministry data bank (339 maternity units). We observed an increase of antenatal care utilization (31 to 42%), the progressive high level of the maternal mortality ratios (245 to 322 per million), intrapartum foetal death (31 to 41%) and neonatal mortality ratios (11 to 12%). We also observed in the regional areas, a positive high variability of unfavourable pregnancy and antenatal care utilization (p=0.00). The authors insist on the improvement of care quality through the reinforcement of caregivers' training followed by extensive research on antenatal care quality. PMID- 12626296 TI - [Current aspects of extra-uterine pregnancy in Libreville (Gabon): an account of 153 cases]. AB - The impact of extra-uterine pregnancy is still increasing in the world. At the same time, its treatment is being improved in developed countries. The purpose of this study is to assess the current aspects of extra-uterine pregnancy in Libreville (Gabon) and to suggest actions which may improve its vital prognosis in the African context. The present study is only prospective and focuses on 153 extra-uterine cases of pregnancy recorded from February 1, 1997 to July 31, 2000. The average frequency was 2.32%. This frequency has been steadily increasing and went from 2% in 1997 to 2.32 % in 2000. The average age of the patients was 29. Extra-uterine pregnancy was diagnosed until the patients were 17. It concerned all parities. The average age was 8 weeks of pregnancy. Three para-clinical investigations allowed the confirmation of the diagnosis: ultrasound scan, coelioscopy, beta GCH plasma dosage. Laparotomy was the main therapeutic solution. No maternal death was recorded. The education of women capable of procreating on the necessity of early consultations, the supplying of hospitals with means which allow para-clinical investigations and the motivation of medical staff seem to be the main solutions to decrease the mortality rate due to extra uterine pregnancy in Africa. PMID- 12626297 TI - [Mucopolysaccharidosis Type 1 (Hurler's disease): the case of a ten-year-old boy]. PMID- 12626301 TI - [Responsibility to be an editor and human, unfairness and supportive forces of the journal]. PMID- 12626298 TI - [RESO (reseau d'entraide pour la sante oculaire), an information network geared to professionally isolated ophthalmologists]. PMID- 12626302 TI - [Stenting of renal artery with or without predilatation in hypertensive patients with renal artery stenosis: results of the nine-months follow-up]. AB - OBJECTIVE: We evaluated the effects of stenting on blood pressure and renal functions in hypertensive patients with proximal/ostial atherosclerotic renal artery stenosis. METHODS: Twenty-six hypertensive patients (9 female, 17 male, mean age 59.0+/-7.4 years) who had renal artery stenosis were included into this study. Their blood pressure, urea and creatinine levels were measured at 24 hours, 3 months and 9 months after procedure. RESULTS: Stents were implanted successfully in all cases. Implanting of stent was done with predilatation in 16 (5%) cases and without predilatation (direct stenting) in 10 (49%) cases. Mean stent diameter and stent length were 7.11+/-0.3 mm and 15.0+/-2.2mm respectively. The antihypertensive drug therapy was stopped in 6 (23%) patients, decreased in 15 patients (58%) and did not change in 5 (19%) patients. No significant changes were observed in urea and creatinine levels. However, creatinine levels were lower after procedure in direct stenting patients than in patients in whom predilatation was applied (0.78+/-0.3 mm; 1.32+/-0.6, p=0.003). This difference disappeared on the 3rd month of follow-up. One patient died during follow-up period. Control angiography was performed in 23 patients. Angiographic restenosis was found in 1 (4.3%) patient. Major events during follow-up period occurred in 2 (8%) patients. CONCLUSION: Stenting in hypertensive patients with renal artery stenosis is reliable and effective procedure. Selection the stenting procedure with or without predilatation depends on the clinical status of patient and the choice of the interventional team. PMID- 12626303 TI - [The efficiency of fast track protocol in elderly patients who underwent coronary artery surgery]. AB - OBJECTIVE: This study is planned to display the efficiency of fast track protocol and its difference from the conventional anesthesia in patients older than 65 years. METHODS: One hundred patients older than 65 years underwent coronary artery surgery between October 2000-March 2001 in cardiovascular surgery clinic were considered in this study. Fifty patients in whom fast track protocol was applied were included into the study group, group A; fifty patients underwent conventional anesthesia technique were referred to the control group, group B. In both groups demographic characteristics, early hospital mortality, operation time, total drainage, number of transfusions, stay in the intensive care unit and discharge time were recorded. RESULTS: The mean age was 69.0+/-3.0 years in group A and 70.4+/-3.6 years in group B. Early hospital mortality was 2% in group A, 10% in group B (p>0.05). Intensive care unit stay was 22.01+/-10.12 hours in group A and 60.18+/-32.23 hours in group B (p<0.05). Discharge time was on 5.5+/ 1.3 day in group A and on 6.9+/-2.3 day in group B (p<0.05). There were no statistical differences between the two groups in respect to other parameters. CONCLUSION: Fast track protocol in patients older than 65 years is a suitable technique by using modern cardiac surgery methods. This protocol is successfully used by selecting the suitable patients and following the patients carefully in the postoperative period. PMID- 12626305 TI - [Evaluation of early alterations in transmitral diastolic flow and tissue Doppler findings of the Basal segments of both ventricles in early period after coronary angioplasty]. AB - OBJECTIVE: Diastolic dysfunction of the left ventricle (LV) appears to be the earliest manifestation of myocardial ischemia. Experimental and clinical studies have shown that both early and late improvements of diastolic function occur after percutaneous transluminal coronary angioplasty (PTCA). Because of the limited utility of transmitral flow profile in the evaluation of the LV diastolic function, recently, assessment of myocardial velocities by Doppler tissue imaging is gaining importance. The aim of this study was to determine the utility of pulsed wave tissue Doppler (PWTD) pattern from the mitral and tricuspid annulus motion in the evaluation of early alterations of the LV diastolic function after revascularization with PTCA in patients with coronary artery disease. METHODS: Pulsed wave tissue Doppler in combination with conventional pulsed-Doppler indices were used to evaluate LV diastolic function before and 24 hours after PTCA. Examinations were performed on 31 patients with chronic ischemic heart disease underwent elective first PTCA. As conventional Doppler indices, early diastolic mitral velocity (e), and its deceleration time (Edt), isovolumic relaxation time (IVRT), late diastolic mitral velocity (a) were measured. Using 2 dimensional echocardiography four chamber apical window, mitral annulus septal and lateral site and tricuspid annulus lateral site were viewed, and PWTD velocity profile were used to calculate; early diastolic maximum velocity and time, late diastolic maximum velocity and time, systolic maximum velocity and time, IVRT and isovolumic contraction time (IVCT) from each site. RESULTS: After PTCA only 2 transmitral conventional pulsed-Doppler indices were changed significantly (IVRT; from 139.7+/-22.2 msec to 120.0+/-15.9 msec, p=0.0001; Edt; from 279+/-11 msec to 248+/-36 msec, p=0.005). However, PWTD (mitral annulus lateral site) analyses showed significant improvement in most of the diastolic parameters: e/a ratio - from 0.80+/-0.26 to 0.89+/-0.22, p=0.012; s wave - from 11.6+/-3.1 cm/sec to 13.2+/-3.6 cm/sec, p=0.03; IVRT - from 130+/-37 msec to 108+/-29 msec, p=0,0001; IVCT - from 84.1+/-19.2 msec to 75.6+/-12.2 msec, p=0.02. Similar significant changes were also observed in the PWTD diastolic parameters of both the mitral annulus septal and tricuspid annulus sites. Peak systolic velocities that reflect the LV systolic functions, of three annular sites significantly improved early after PTCA, however ejection fraction was not changed as much as tissue Doppler parameters (s maximum velocity before PTCA: 11.7+/-3.1 cm/sec versus 13.2+/-3.6 cm/sec after PTCA, p=0.03). CONCLUSION: Tissue Doppler indices of the mitral annulus reflecting both the diastolic and systolic functions, improve early after successful PTCA in patients with coronary artery disease. PMID- 12626309 TI - The effects of handgrip stress test on hemodynamic parameters before and after cilazapril treatment in patients with heart failure. AB - OBJECTIVE: To assess the effect of cilazapril treatment on several hemodynamic parameters during handgrip maneuvers in patients with congestive heart failure. Cilazapril, an ACE inhibitor with high affinity, has been shown to be highly effective against a variety of vascular disorders. The effectiveness of isometric handgrip exercise on changes of cardiovascular hemodynamic parameters before and after cilazapril treatment in patients with congestive heart failure is unknown. METHODS: The study population included 30 patients (16 male, 14 female) with mean age of 65+/-18 years. The effects of handgrip maneuver on hemodynamic parameters were studied by right heart catheterization and Doppler echocardiography. RESULTS: Heart rate (HR) and mean arterial pressures (MAP) increased significantly after handgrip maneuver (from 95+/-6 beats/min to 101+/-12 beats/min; from 109+/-15 mm Hg to 118+/-19 mm Hg, p<0.05 respectively). Pulmonary capillary wedge pressure (PCWP), pulmonary artery systolic (s) and diastolic (d) pressures (PAP), cardiac index (CI), right ventricular systolic and diastolic pressures (RVPs and RVPd), left ventricular ejection fraction (LVEF), right ventricular ejection fraction (RVEF) did not change after handgrip maneuvers (p>0.05). On the other hand, PAPs and PAPd, RVPs and RVPd, MAP and HR (p<0.05) decreased significantly during handgrip maneuvers after cilazapril treatment. However PCWP and CI, LVEF, RVEF did not change after treatment (p>0.05). CONCLUSION: Cardiovascular response to handgrip maneuver may be a marker of failure to respond to compensatory mechanisms. Cilazapril treatment was associated with significant improvement in hemodynamic parameters during handgrip stress test, the mechanisms of which are increased sympathetic and renin angiotensin system activation, and altered vascular tonus. PMID- 12626307 TI - The association of plasma homocysteine, coronary risk factors and serum nitrite in patients with coronary artery disease, vascular syndrome x and healthy subjects. AB - OBJECTIVE: We evaluated the association of plasma total homocysteine (tHcy), cardiac risk factors and total nitrite in coronary artery disease (CAD) patients, cardiac syndrome X patients and in healthy subjects. METHODS: Forty two CAD, 22 cardiac syndrome X patients and 30 healthy subjects, aged 30 to 75 years were included into the study. Blood samples of tHcy, serum total nitrite and cardiac risk factors were studied appropriately. The results were compared between the groups. The independent contributions of tHcy and total nitrite to CAD and cardiac syndrome X and their interactions with cardiac risk factors were evaluated. RESULTS: After adjusting for age, median values of tHcy and total nitrite were evaluated for their skewness. Coronary artery disease patients had higher median plasma tHcy levels than cardiac syndrome X patients (p<0.001) and healthy subjects (p<0.001) and lower serum total nitrite levels than patients in the two other groups (p<0.05), respectively. Using a univariate linear regression analysis tHcy had a moderately significant positive correlation with age (beta=0.34, p=0.002) and a weakly significant inverse correlation with female gender (beta=-0.24, p=0.032). Using a partial correlation analysis by controlling for age, gender and clinical situations tHcy had a positive but moderately significant correlation with LDL cholesterol (r=0.23, p=0.01) and triglycerides (r=0.27, p=0.016). Total nitrite had a positive but weakly significant correlation with HDL cholesterol (r=0.23, p=0.04) and fibrinogen (r=0.24, p=0.03) and an inverse but moderately significant correlation with LDL cholesterol (r= 0.37,p=0.001). Using a multivariate stepwise regression analysis total nitrite was inversely and significantly associated with tHcy (beta=-0.45) in the control group. The contribution of HDL cholesterol to the association was beta=-0.45, p=0.044, R2=36.2%, HDL cholesterol with fibrinogen--beta=-0.45, p=0.05, R2=36.6% and HDL cholesterol with LDL cholesterol--beta=-0.45, p=0.05, R2=36.3%. In a forward stepwise logistic regression analysis the age adjusted odds ratio (OR) for coronary artery disease per standard deviation change in log-transformed tHcy concentration was - 0.82, p=0.013 and in total nitrite concentration was - 1.08, p=0.02. Using the same model neither tHcy nor total nitrite was associated with cardiac syndrome X (p=0.221 and p=0.112), respectively. CONCLUSION: The low nitrite levels can be a marker of endothelial dysfunction in the presence of hyperhomocysteinemia and other cardiac risk factors. Our results might support endothelial dysfunction in CAD but not in cardiac syndrome X patients. PMID- 12626310 TI - Noninvasive electrocardiographic findings and plasma norepinephrine levels in patients with post-myocardial infarction receiving anti-anginal agents. AB - OBJECTIVE: The aim of this study was to investigate the effects of anti-anginal agents on plasma norepinephrine (NE) levels and the autonomic nerve functions evaluated by advanced noninvasive electrocardiographic (ECG) tests in post myocardial infarction (PMI) patients. METHODS: The subjects were 89 PMI patients who had suffered myocardial infarction (MI) at least 2 months before they participated in this study, and who had been taking anti-anginal agent mono therapy (typical Japanese doses) for at least 6 months. Subjects were classified into the following 3 groups, based on type of anti-anginal agent: calcium antagonists (n=31, 60+/-12 years), nitrates (n=29, 56+/-11 years) and beta blockers (n=29, 63+/-14 years). Left ventricular late potentials (LP), heart rate variability (HRV), T wave alternans (TWA), QT dispersion (QTd), and plasma NE levels of all subjects were assessed. There were no significant differences in age, gender, MI location or coronary risk factors between the 3 groups. RESULTS: There were no significant differences in the number of subjects who satisfied criteria for LP, TWA, and QTd between the 3 groups. Mean high frequency power of HRV of the calcium antagonist group was significantly (p<0.05) lower than those of the nitrate and beta-blocker groups. All 3 groups had similar LF/HF, TWA microvoltage and QTd values, but mean plasma NE level of the calcium antagonist group was significantly (p<0.01) higher than those of the nitrate and beta blocker groups. CONCLUSION: These results indicate that calcium antagonist therapy in PMI patients lowers parasympathetic tone and elevates plasma NE levels. However, in the present study, these values remained within normal ranges. PMID- 12626311 TI - [Myocardial noncompaction: two cases and review]. AB - Myocardial noncompaction (MN) is a rare form of cardiomyopathy due to an arrest in endomyocardial embryogenesis. It is characterized by multiple prominent trabeculations with deep intertrabecular recesses in ventricular walls. The disease usually affects the left ventricle and may cause cardiac failure, arrhythmias, and systemic embolism. We would like to present two rare cases of MN with a poor long-term prognosis. The first case is an unique case in the literature having bioprosthetic valve implantation and associated with a recovery of heart failure after the operation, the second case is another interesting case because of the association of A-V complete block, and multiple structural cardiac abnormalities. PMID- 12626312 TI - [Pulsed wave tissue Doppler echocardiography]. AB - Tissue Doppler imaging is a recently developed technique that allows direct measurement of regional or global systolic and diastolic myocardial motion velocities. Myocardial velocities may be recorded in the pulsed wave tissue Doppler (PWTD) or color tissue Doppler. Pulsed wave tissue Doppler appears to be relatively independent of preload and provides important information in various heart diseases. Technical basis, normal pattern and clinical utility of PWTD were reviewed in this article. PMID- 12626313 TI - [Mitral valve prolapse syndrome: orthostatic hypotension and physiopathology of its clinical symptomathologies]. AB - Although several investigations on mitral valve prolapse syndrome (MVPS) have been performed, clinical symptoms of this syndrome are not yet clarified. Atypical chest pain, palpitations, fatigue, dyspnea and anxiety are the most frequent symptoms associated with this syndrome. However, dizziness and syncope may be serious symptoms in MVPS. Dizziness and syncope are related to cardiac arrhythmias and are proposed to distinguish types, frequency of arrhythmias and relation to the symptoms. Orthostatic hypotension and tachycardia rarely occur in MVPS. The physiopathological mechanisms of these symptoms are not known clearly, but multifactorial causes are thought to be responsible including autonomic dysfunction, hyperadrenergic state, abnormalities in regulation of baroreceptors, parasympathetic derangements, decrease of intravascular volume, abnormal renin aldosterone response to depletion of intravascular volume and abnormal release of atrial natriuretic factor. PMID- 12626314 TI - [Diuretics use in edematous patients]. PMID- 12626315 TI - [Congenital bicuspid aorta associated with ascending aortic aneurysm]. PMID- 12626317 TI - [Extraction of the infected defibrillator lead using excimer laser system]. PMID- 12626316 TI - Bilateral renal arterial embolisation in a patient with mitral stenosis and atrial fibrillation: an uncommon reason of flank pain. PMID- 12626318 TI - [Myeloid leukaemia case with myocardial infarction and septal rupture secondary to infective endocarditis]. PMID- 12626319 TI - [A case of primary malignant cardiac tumour considered as a pericardial tuberculosis]. PMID- 12626320 TI - [Recurrent angina due to coronary-subclavian steal syndrome]. PMID- 12626321 TI - Altered energetic properties in skeletal muscle of men with well-controlled insulin-dependent (type 1) diabetes. AB - This study asked whether the energetic properties of muscles are changed by insulin-dependent diabetes mellitus (or type 1 diabetes), as occurs in obesity and type 2 diabetes. We used (31)P magnetic resonance spectroscopy to measure glycolytic flux, oxidative flux, and contractile cost in the ankle dorsiflexor muscles of 10 men with well-managed type 1 diabetes and 10 age- and activity matched control subjects. Each subject performed sustained isometric muscle contractions lasting 30 and 120 s while attempting to maintain 70-75% of maximal voluntary contraction force. An altered glycolytic flux in type 1 diabetic subjects relative to control subjects was apparent from significant differences in pH in muscle at rest and at the end of the 120-s bout. Glycolytic flux during exercise began earlier and reached a higher peak rate in diabetic patients than in control subjects. A reduced oxidative capacity in the diabetic patients' muscles was evident from a significantly slower phosphocreatine recovery from a 30-s exercise bout. Our findings represent the first characterization of the energetic properties of muscle from type 1 diabetic patients. The observed changes in glycolytic and oxidative fluxes suggest a diabetes-induced shift in the metabolic profile of muscle, consistent with studies of obesity and type 2 diabetes that point to common muscle adaptations in these diseases. PMID- 12626322 TI - Protein tyrosine phosphatases: the quest for negative regulators of insulin action. AB - Type 2 diabetes is increasing at an alarming rate worldwide, and there has been a considerable effort in several laboratories to identify suitable targets for the design of drugs against the disease. To this end, the protein tyrosine phosphatases that attenuate insulin signaling by dephosphorylating the insulin receptor (IR) have been actively pursued. This is because inhibiting the phosphatases would be expected to prolong insulin signaling and thereby facilitate glucose uptake and, presumably, result in a lowering of blood glucose. Targeting the IR protein tyrosine phosphatase, therefore, has the potential to be a significant disease-modifying strategy. Several protein tyrosine phosphatases (PTPs) have been implicated in the dephosphorylation of the IR. These phosphatases include PTPalpha, LAR, CD45, PTPepsilon, SHP2, and PTP1B. In most cases, there is evidence for and against the involvement of the phosphatases in insulin signaling. The most convincing data, however, support a critical role for PTP1B in insulin action. PTP1B knockout mice are not only insulin sensitive but also maintain euglycemia (in the fed state), with one-half the level of insulin observed in wild-type littermates. Interestingly, these mice are also resistant to diet-induced obesity when fed a high-fat diet. The insulin-sensitive phenotype of the PTP1B knockout mouse is reproduced when the phosphatase is also knocked down with an antisense oligonucleotide in obese mice. Thus PTP1B appears to be a very attractive candidate for the design of drugs for type 2 diabetes and obesity. PMID- 12626323 TI - Glucagon and regulation of glucose metabolism. AB - As a counterregulatory hormone for insulin, glucagon plays a critical role in maintaining glucose homeostasis in vivo in both animals and humans. To increase blood glucose, glucagon promotes hepatic glucose output by increasing glycogenolysis and gluconeogenesis and by decreasing glycogenesis and glycolysis in a concerted fashion via multiple mechanisms. Compared with healthy subjects, diabetic patients and animals have abnormal secretion of not only insulin but also glucagon. Hyperglucagonemia and altered insulin-to-glucagon ratios play important roles in initiating and maintaining pathological hyperglycemic states. Not surprisingly, glucagon and glucagon receptor have been pursued extensively in recent years as potential targets for the therapeutic treatment of diabetes. PMID- 12626324 TI - Interaction of glucagon and epinephrine in the control of hepatic glucose production in the conscious dog. AB - Epinephrine increases net hepatic glucose output (NHGO) mainly via increased gluconeogenesis, whereas glucagon increases NHGO mainly via increased glycogenolysis. The aim of the present study was to determine how the two hormones interact in controlling glucose production. In 18-h-fasted conscious dogs, a pancreatic clamp initially fixed insulin and glucagon at basal levels, following which one of four protocols was instituted. In G + E, glucagon (1.5 ng x kg(-1) x min(-1); portally) and epinephrine (50 ng x kg(-1) x min(-1); peripherally) were increased; in G, glucagon was increased alone; in E, epinephrine was increased alone; and in C, neither was increased. In G, E, and C, glucose was infused to match the hyperglycemia seen in G + E ( approximately 250 mg/dl). The areas under the curve for the increase in NHGO, after the change in C was subtracted, were as follows: G = 661 +/- 185, E = 424 +/- 158, G + E = 1178 +/- 57 mg/kg. Therefore, the overall effects of the two hormones on NHGO were additive. Additionally, glucagon exerted its full glycogenolytic effect, whereas epinephrine exerted its full gluconeogenic effect, such that both processes increased significantly during concurrent hormone administration. PMID- 12626326 TI - Chemical sympathectomy alters regulation of body weight during prolonged ICV leptin infusion. AB - To assess the importance of the sympathetic nervous system in regulating body weight during prolonged leptin infusion, we evaluated food intake, body weight, and physical activity in conscious, unrestrained rats. Initial studies illustrated that prolonged intracerebroventricular (ICV) infusion of leptin enhanced substrate oxidation so that adipose tissue lipid stores were completely ablated, and muscle triglyceride and liver glycogen stores were depleted. After neonatal chemical sympathectomy, changes in weight and food intake were compared in groups of sympathectomized (SYM) and control (CON) adult animals during ICV infusion of leptin. CON animals lost 60 +/- 9 g over 10 days vs. 25 +/- 3 g in the SYM animals when food intake was matched between the two groups. Greater weight loss despite similar energy intake points to an important role of the sympathetic nervous system in stimulating energy expenditure during ICV leptin infusion by increasing the resting metabolic rate, since no differences in physical activity were observed between CON and SYM groups. In conclusion, activation of the SNS by leptin increases energy expenditure by augmenting the resting metabolic rate. PMID- 12626325 TI - Skeletal muscle lipid metabolism with obesity. AB - The objectives of this study were to 1). examine skeletal muscle fatty acid oxidation in individuals with varying degrees of adiposity and 2). determine the relationship between skeletal muscle fatty acid oxidation and the accumulation of long-chain fatty acyl-CoAs. Muscle was obtained from normal-weight [n = 8; body mass index (BMI) 23.8 +/- 0.58 kg/m(2)], overweight/obese (n = 8; BMI 30.2 +/- 0.81 kg/m(2)), and extremely obese (n = 8; BMI 53.8 +/- 3.5 kg/m(2)) females undergoing abdominal surgery. Skeletal muscle fatty acid oxidation was assessed in intact muscle strips. Long-chain fatty acyl-CoA concentrations were measured in a separate portion of the same muscle tissue in which fatty acid oxidation was determined. Palmitate oxidation was 58 and 83% lower in skeletal muscle from extremely obese (44.9 +/- 5.2 nmol x g(-1) x h(-1)) patients compared with normal weight (71.0 +/- 5.0 nmol x g(-1) x h(-1)) and overweight/obese (82.2 +/- 8.7 nmol x g(-1) x h(-1)) patients, respectively. Palmitate oxidation was negatively (R = -0.44, P = 0.003) associated with BMI. Long-chain fatty acyl-CoA content was higher in both the overweight/obese and extremely obese patients compared with normal-weight patients, despite significantly lower fatty acid oxidation only in the extremely obese. No associations were observed between long-chain fatty acyl CoA content and palmitate oxidation. These data suggest that there is a defect in skeletal muscle fatty acid oxidation with extreme obesity but not overweight/obesity and that the accumulation of intramyocellular long-chain fatty acyl-CoAs is not solely a result of reduced fatty acid oxidation. PMID- 12626327 TI - Functional and morphological evidence for a ventricular conduction system in zebrafish and Xenopus hearts. AB - Zebrafish and Xenopus have become popular model organisms for studying vertebrate development of many organ systems, including the heart. However, it is not clear whether the single ventricular hearts of these species possess any equivalent of the specialized ventricular conduction system found in higher vertebrates. Isolated hearts of adult zebrafish (Danio rerio) and African toads (Xenopus laevis) were stained with voltage-sensitive dye and optically mapped in spontaneous and paced rhythms followed by histological examination focusing on myocardial continuity between the atrium and the ventricle. Spread of the excitation wave through the atria was uniform with average activation times of 20 +/- 2 and 50 +/- 2 ms for zebrafish and Xenopus toads, respectively. After a delay of 47 +/- 8 and 414 +/- 16 ms, the ventricle became activated first in the apical region. Ectopic ventricular activation was propagated significantly more slowly (total ventricular activation times: 24 +/- 3 vs. 14 +/- 2 ms in zebrafish and 74 +/- 14 vs. 35 +/- 9 ms in Xenopus). Although we did not observe any histologically defined tracts of specialized conduction cells within the ventricle, there were trabecular bands with prominent polysialic acid-neural cell adhesion molecule staining forming direct myocardial continuity between the atrioventricular canal and the apex of the ventricle; i.e., the site of the epicardial breakthrough. We thus conclude that these hearts are able to achieve the apex-to-base ventricular activation pattern observed in higher vertebrates in the apparent absence of differentiated conduction fascicles, suggesting that the ventricular trabeculae serve as a functional equivalent of the His-Purkinje system. PMID- 12626329 TI - Failing atrial myocardium: energetic deficits accompany structural remodeling and electrical instability. AB - The failing ventricular myocardium is characterized by reduction of high-energy phosphates and reduced activity of the phosphotransfer enzymes creatine kinase (CK) and adenylate kinase (AK), which are responsible for transfer of high-energy phosphoryls from sites of production to sites of utilization, thereby compromising excitation-contraction coupling. In humans with chronic atrial fibrillation (AF) unassociated with congestive heart failure (CHF), impairment of atrial myofibrillar energetics linked to oxidative modification of myofibrillar CK has been observed. However, the bioenergetic status of the failing atrial myocardium and its potential contribution to atrial electrical instability in CHF have not been determined. Dogs with (n = 6) and without (n = 6) rapid pacing induced CHF underwent echocardiography (conscious) and electrophysiological (under anesthesia) studies. CHF dogs had more pronounced mitral regurgitation, higher atrial pressure, larger atrial area, and increased atrial fibrosis. An enhanced propensity to sustain AF was observed in CHF, despite significant increases in atrial effective refractory period and wavelength. Profound deficits in atrial bioenergetics were present with reduced activities of the phosphotransfer enzymes CK and AK, depletion of high-energy phosphates (ATP and creatine phosphate), and reduction of cellular energetic potential (ATP-to-ADP and creatine phosphate-to-Cr ratios). AF duration correlated with left atrial area (r = 0.73, P = 0.01) and inversely with atrial ATP concentration (r = -0.75, P = 0.005), CK activity (r = -0.57, P = 0.054), and AK activity (r = -0.64, P = 0.02). Atrial levels of malondialdehyde, a marker of oxidative stress, were significantly increased in CHF. Myocardial bioenergetic deficits are a conserved feature of dysfunctional atrial and ventricular myocardium in CHF and may constitute a component of the substrate for AF in CHF. PMID- 12626328 TI - Role of PKC in autocrine regulation of rat ventricular K+ currents by angiotensin and endothelin. AB - Transient and sustained K(+) currents were measured in isolated rat ventricular myocytes obtained from control, steptozotocin-induced (Type 1) diabetic, and hypothyroid rats. Both currents, attenuated by the endocrine abnormalities, were significantly augmented by in vitro incubation (>6 h) with the angiotensin converting enzyme inhibitor quinapril or the angiotensin II (ANG II) receptor blocker saralasin. Western blots indicated a parallel increase in Kv4.2 and Kv1.2, channel proteins that underlie the transient and (part of the) sustained currents. Under diabetic and hypothyroid conditions, both currents were also augmented by an endothelin receptor blocker (PD142893) or by an endothelin converting enzyme inhibitor. Kv4.2 density was also enhanced by PD142893. Incubation (>5 h) with the PKC inhibitor bis-indolylmaleimide augmented both currents, whereas the PKC activator dioctanoyl-rac-glycerol (DiC8) prevented the augmentation of currents by quinapril. DiC8 also prevented the augmentation of Kv4.2 density by quinapril. Specific peptides that activate PKC translocation indicated that PKC-epsilon and not PKC-delta is involved in ANG II action on these currents. In control myocytes, quinapril and PD142893 augmented the sustained late current but had no effect on peak current. It is concluded that an autocrine release of angiotensin and endothelin in diabetic and hypothyroid conditions attenuates K(+) currents by suppressing the synthesis of some K(+) channel proteins, with the effects mediated at least partially by PKC-epsilon. PMID- 12626330 TI - Side population cells and Bcrp1 expression in lung. AB - Side population (SP) cells are a rare subset of cells found in various tissues that are highly enriched for stem cell activity. SP cells can be isolated by dual wavelength flow cytometry because of their capacity to efflux Hoechst dye, a process mediated by the ATP-binding cassette transporter breast cancer resistance protein (Bcrp) 1. By performing flow cytometry of enzymedigested mouse lung stained with Hoechst dye, we found that SP cells comprise 0.03-0.07% of total lung cells and are evenly distributed in proximal and distal lung regions. By RT PCR, we found that lung SP cells express hepatocyte nuclear factor-3beta, but not thyroid transcription factor-1. Surface marker analysis revealed lung SP cells to be stem cell antigen 1 positive, Bcrp1 positive, lineage marker negative, and heterogeneous at the CD45 locus. As expected, we did not detect lung SP cells in Bcrp1-deficient animals. We, therefore, employed nonisotopic in situ hybridization and immunostaining for Bcrp1 as a strategy to localize these cells in vivo. Expression was observed in distinct lung cell types: bronchial and vascular smooth muscle cells and round cells within the distal air space. We confirmed the expression of Bcrp1 in primary bronchial smooth muscle cell cultures (BSMC) and in lavaged distal airway cells, but neither possessed the capacity to efflux Hoechst dye. In BSMC, Bcrp1 was localized to an intracellular compartment, suggesting that the molecular site of Bcrp1 expression regulates SP phenotype. PMID- 12626331 TI - Effects of hyperoxia on VEGF, its receptors, and HIF-2alpha in the newborn rat lung. AB - Signaling through the hypoxia inducible factor (HIF)-VEGF-VEGF receptor system (VEGF signaling system) leads to angiogenesis and epithelial cell proliferation and is a key mechanism regulating alveolarization in lungs of newborn rats. Hyperoxia exposure (>95% O2 days 4-14) arrests lung alveolarization and may do so through suppression of the VEGF signaling system. Lung tissue mRNA levels of HIF 2alpha and VEGF increased from days 4-14 in normoxic animals, but hyperoxia suppressed these increases. Levels of HIF-2alpha and VEGF mRNA were correlated in the air but not the O2-treated group, suggesting that the low levels of HIF 2alpha observed at high O2 concentrations are not stimulating VEGF expression. VEGF164 protein levels increased with developmental age, and with hyperoxia to day 9, but continuing hyperoxia decreased levels by day 12. VEGFR1 and VEGFR2 mRNA expression also increased in air-exposed animals, and these, too, were significantly decreased by hyperoxia by day 9 and day 12, respectively. Receptor protein levels did not increase with development; however, O2 did decrease protein to less than air values. Hyperoxic suppression of VEGF signaling from days 9-14 may be one mechanism by which alveolarization is arrested. PMID- 12626333 TI - Effect of poly(ADP ribose) synthetase inhibition on burn and smoke inhalation injury in sheep. AB - We investigated the role of the nuclear enzyme poly (ADP ribose) synthetase (PARS) in the pathogenesis of combined burn and smoke inhalation (burn/smoke) injury in an ovine model. Eighteen sheep were operatively prepared for chronic study. PARS inhibition was achieved by treatment with a novel and selective PARS inhibitor INO-1001. The PARS inhibitor attenuated 1) lung edema formation, 2) deterioration of gas exchange, 3) changes in airway blood flow, 4) changes in airway pressure, 5) lung histological injury, and 6) systemic vascular leakage. Lipid oxidation and plasma nitrite/nitrate (stable breakdown products of nitric oxide) levels were suppressed with the use of INO-1001. We conclude that PARS inhibition attenuates various aspects of the pathophysiological response in a clinically relevant experimental model of burn/smoke inhalation injury. PMID- 12626332 TI - Role of sphingosine-1 phosphate in the enhancement of endothelial barrier integrity by platelet-released products. AB - In vitro and in vivo evidence indicates that circulating platelets affect both vascular integrity and hemostasis. How platelets enhance the permeability barrier of the vascular endothelium is not well understood. We measured the effect of isolated human platelets on human pulmonary artery endothelial cell (EC) barrier integrity by monitoring transmonolayer electrical resistance. EC barrier function was significantly increased by the addition of platelets ( approximately 40% maximum, 2.5 x 106 platelets/ml). Platelet supernatants, derived from 2.5 x 106 platelets/ml, reproduced the barrier enhancement and reversed the barrier dysfunction produced by the edemagenic agonist thrombin, which implicates a soluble barrier-promoting factor. The barrier-enhancing effect of platelet supernatants was heat stable but was attenuated by either charcoal delipidation (suggesting a vasoactive lipid mediator) or pertussis toxin, implying involvement of a Gialpha-coupled receptor signal transduction pathway. Sphingosine-1 phosphate (S1P), a sphingolipid that is released from activated platelets, is known to ligate G protein-coupled EC differentiation gene (EDG) receptors, increase EC electrical resistance, and reorganize the actin cytoskeleton (Garcia JG, Liu F, Verin AD, Birukova A, Dechert MA, Gerthoffer WT, Bamberg JR, and English D. J Clin Invest 108: 689-701, 2001). Infection of EC with an adenoviral vector expressing an antisense oligonucleotide directed against EDG-1 but not infection with control vector attenuated the barrier-enhancing effect of both platelet supernatants and S1P. These results indicate that a major physiologically relevant vascular barrier-protective mediator produced by human platelets is S1P. PMID- 12626334 TI - Targeted blocking of gene expression for CGRP receptors elevates pulmonary artery pressure in hypoxic rats. AB - We previously described the protection by calcitonin gene-related peptide (CGRP) against hypoxic pulmonary hypertension. Here, we examine the roles of its putative receptor RDC-1 and receptor activity-modifying protein (RAMP) 1 in mediating this protection by selectively inhibiting their synthesis. RAMP1 is an accessory protein for another putative CGRP receptor, calcitonin receptor-like receptor. Antisense oligodeoxyribonucleotides (ASODNs, 5 mg.kg-1.day-1 or 5 and 10 mg.kg-1.day-1 for RDC-1) targeting RAMP1 and RDC-1 mRNAs were chronically infused to the pulmonary circulation of male Sprague-Dawley rats during 7 days of normoxia or hypobaric hypoxia (380 mmHg), and alpha-CGRP ASODN was used as a technical control. CGRP, RAMP1, and RDC-1 ASODNs significantly elevated pulmonary artery pressure (PPA) in chronic hypoxic rats compared with hypoxic mismatched ASODN (MMODN) and saline vehicle controls. CGRP and RAMP1 ASODNs raised PPA in normoxic rats briefly exposed to 10% O2 above MMODN and saline controls. Moreover, normoxic rats treated with CGRP ASODN had higher basal pulmonary vascular tone compared with controls. These data confirm the protective role of CGRP in the pulmonary circulation and suggest that endogenous RAMP1 and RDC-1 are essential in regulation of PPA in hypoxia. This is the first in vivo evidence supporting RDC-1 and RAMP1 as functional CGRP receptor and receptor component. PMID- 12626335 TI - Allergen-induced airway disease is mouse strain dependent. AB - We investigated the development of airway hyperreactivity (AHR) and inflammation in the lungs of nine genetically diverse inbred strains of mice [129/SvIm, A/J, BALB/cJ, BTBR+(T)/tf/tf, CAST/Ei, C3H/HeJ, C57BL/6J, DBA/2J, and FVB/NJ] after sensitization and challenge with ovalbumin (OVA). At 24, 48, and 72 h post-OVA exposure, the severity of AHR and eosinophilic inflammation of the mouse strains ranged from relatively unresponsive to responsive. The severity of the airway eosinophilia of some strains did not clearly correlate with the development of AHR. The temporal presence of T helper type 2 cytokines in lung lavage fluid also varied markedly among the strains. The levels of IL-4 and IL-13 were generally increased in the strains with the highest airway eosinophilia at 24 and 72 h postexposure, respectively; the levels of IL-5 were significantly increased in most of the strains with airway inflammation over the 72-h time period. The differences of physiological and biological responses among the inbred mouse strains after OVA sensitization and challenge support the hypothesis that genetic factors contribute, in part, to the development of allergen-induced airway disease. PMID- 12626336 TI - Pulmonary vascular dysfunction in preterm lambs with chronic lung disease. AB - Chronic lung injury from prolonged mechanical ventilation after premature birth inhibits the normal postnatal decrease in pulmonary vascular resistance (PVR) and leads to structural abnormalities of the lung circulation in newborn sheep. Compared with normal lambs born at term, chronically ventilated preterm lambs have increased pulmonary arterial smooth muscle and elastin, fewer lung microvessels, and reduced abundance of endothelial nitric oxide synthase. These abnormalities may contribute to impaired respiratory gas exchange that often exists in infants with chronic lung disease (CLD). Nitric oxide inhalation (iNO) reduces PVR in human infants and lambs with persistent pulmonary hypertension. We wondered whether iNO might have a similar effect in lambs with CLD. We therefore studied the effect of iNO on PVR in lambs that were delivered prematurely at approximately 125 days of gestation (term = 147 days) and mechanically ventilated for 3 wk. All of the lambs had chronically implanted catheters for measurement of pulmonary vascular pressures and blood flow. During week 2 of mechanical ventilation, iNO at 15 parts/million for 1 h decreased PVR by approximately 20% in 12 lambs with evolving CLD. When the same study was repeated in eight lambs at the end of week 3, iNO had no significant effect on PVR. To see whether this loss of iNO effect on PVR might reflect dysfunction of lung vascular smooth muscle, we infused 8-bromo-guanosine 3',5'-cyclic monophosphate (cGMP; 150 microg. kg(-1). min(-1) iv) for 15-30 min in four of these lambs at the end of week 3. PVR consistently decreased by 30-35%. Lung immunohistochemistry and immunoblot analysis of excised pulmonary arteries from lambs with CLD, compared with control term lambs, showed decreased soluble guanylate cyclase (sGC). These results suggest that loss of pulmonary vascular responsiveness to iNO in preterm lambs with CLD results from impaired signaling, possibly related to deficient or defective activation of sGC, the intermediary enzyme through which iNO induces increased vascular smooth muscle cell cGMP and resultant vasodilation. PMID- 12626337 TI - Protein tyrosine phosphatase activity regulates endothelial cell-cell interactions, the paracellular pathway, and capillary tube stability. AB - Protein tyrosine phosphorylation is tightly regulated through the actions of both protein tyrosine kinases and protein tyrosine phosphatases. In this study, we demonstrate that protein tyrosine phosphatase inhibition promotes tyrosine phosphorylation of endothelial cell-cell adherens junction proteins, opens an endothelial paracellular pathway, and increases both transendothelial albumin flux and neutrophil migration. Tyrosine phosphatase inhibition with sodium orthovanadate or phenylarsine oxide induced dose- and time-dependent increases in [14C]bovine serum albumin flux across postconfluent bovine pulmonary artery endothelial cell monolayers. These increases in albumin flux were coincident with actin reorganization and intercellular gap formation in both postconfluent monolayers and preformed endothelial cell capillary tubes. Vanadate (25 microM) increased tyrosine phosphorylation of endothelial cell proteins 12-fold within 1 h. Tyrosine phosphorylated proteins were immunolocalized to the intercellular boundaries, and several were identified as the endothelial cell-cell adherens junction proteins, vascular-endothelial cadherin, and beta-, gamma-, and p120 catenin as well as platelet endothelial cell adhesion molecule-1. Of note, these tyrosine phosphorylation events were not associated with disassembly of the adherens junction complex or its uncoupling from the actin cytoskeleton. The dose and time requirements for vanadate-induced increases in phosphorylation were comparable with those defined for increments in transendothelial [14C]albumin flux and neutrophil migration, and pretreatment with the tyrosine kinase inhibitor herbimycin A protected against these effects. These data suggest that protein tyrosine phosphatases and their substrates, which localize to the endothelial cell-cell boundaries, regulate adherens junctional integrity, the movement of macromolecules and cells through the endothelial paracellular pathway, and capillary tube stability. PMID- 12626338 TI - Mechanisms of alternative pre-messenger RNA splicing. AB - Alternative pre-mRNA splicing is a central mode of genetic regulation in higher eukaryotes. Variability in splicing patterns is a major source of protein diversity from the genome. In this review, I describe what is currently known of the molecular mechanisms that control changes in splice site choice. I start with the best-characterized systems from the Drosophila sex determination pathway, and then describe the regulators of other systems about whose mechanisms there is some data. How these regulators are combined into complex systems of tissue specific splicing is discussed. In conclusion, very recent studies are presented that point to new directions for understanding alternative splicing and its mechanisms. PMID- 12626339 TI - Semisynthesis of proteins by expressed protein ligation. AB - Expressed protein ligation (EPL) is a protein engineering approach that allows recombinant and synthetic polypeptides to be chemoselectively and regioselectively joined together. The approach makes the primary structure of most proteins accessible to the tools of synthetic organic chemistry, enabling the covalent structure of proteins to be modified in an unprecedented fashion. The ability to incorporate noncoded amino acids, biophysical probes, and stable isotopes into specific locations within proteins provides research tools to peer into the inner workings of these molecules. In this review I discuss the development of this technology, its broad application to biological systems, and its possible role in the area of proteomics. PMID- 12626340 TI - Effects of endothelin-1 on epithelial ion transport in human airways. AB - Endothelin-1 (ET-1) exerts many biological effects in airways, including bronchoconstriction, airway mucus secretion, cell proliferation, and inflammation. We investigated the effect of ET-1 on Na absorption and Cl secretion in human bronchial epithelial cells. Addition of 10(-7) M ET-1 had no effect on the inhibition of the short circuit current (Isc) induced by amiloride, a Na channel blocker. Addition of 10(-7) M ET-1 to the apical bath in the presence of amiloride increased Isc in cultured human bronchial epithelial cells studied in Ussing chambers. No effect was observed when ET-1 was added to basolateral bath, indicating that the involved ET-1 receptors are likely present only in the apical membrane of the cells. Use of Cl-free solutions and bumetanide reduced the ET-1-induced increases in Isc, indicating that ET-1 stimulates Cl secretion. The ET-1-induced increase in Isc was prevented by exposure to the ETB receptor antagonist BQ-788 but not to the ETA receptor antagonist BQ-123. ET-1 did not raise intracellular Ca levels, but increased the intracellular concentration of cAMP. These findings indicate that ET-1 is a Cl secretagogue in human airways and acts presumably through apically located ETB receptors and activation of the cAMP pathway. PMID- 12626341 TI - Lactoperoxidase and human airway host defense. AB - The lactoperoxidase (LPO) antibiotic system is a well-characterized component of mammary and salivary gland secretions. Because LPO has been shown to function in ovine airways, human airway tissue and secretions were examined for the presence of LPO and its substrate, the anion thiocyanate (SCN-). In addition, human airway secretions were tested for LPO-mediated antibacterial activity, and LPO's activity was assessed against some human airway pathogens. The data showed that normal human airway secretions contained LPO enzyme activity (0.65 +/- 0.09 microg/mg secreted protein; n = 17), and Western blots of secretions demonstrated bands of the expected sizes for LPO. LPO mRNA was detected in trachea by sequencing PCR-amplified cDNA. SCN-, LPO's substrate, was present in undiluted airway secretions at concentrations sufficient for LPO catalysis (0.46 +/- 0.19 mM; n = 8), and diluted secretions contained antibacterial activity with LPO-like properties. Immunocytochemistry localized LPO to submucosal glands in human bronchi. Finally, as expected based on the known antibacterial spectrum of the LPO system, airway secretions showed LPO-dependent activity against Pseudomonas aeruginosa. In addition, the airway LPO system was shown to be effective against Burkholderia cepacia and Haemophilus influenzae. Thus, a functional LPO system exists in human airways and may contribute to airway host defense against infection. PMID- 12626342 TI - Asbestos-induced apoptosis is protein kinase C delta-dependent. AB - Alveolar epithelial and mesothelial cells undergo apoptosis in response to asbestos, a phenomenon that may be important in injury and/or initiation of compensatory proliferation. Here, we report a functional role of protein kinase (PKC)delta in apoptosis by crocidolite asbestos. We first show that asbestos increases the kinase activity of PKC delta in alveolar type II epithelial cells (C10 line) and causes its translocation to mitochondria, events associated with caspase-9 cleavage and apoptosis as detected by the Apostain technique. Pretreatment of C10 cells with rottlerin (Rot), a PKC delta-selective inhibitor, before addition of asbestos prevented cleavage of caspase-9 and blocked the appearance of apoptotic cells. Asbestos-induced apoptosis also was inhibited in cells stably expressing a dominant-negative kinase-deficient mutant of PKC delta (dnPKC delta), but not dnPKC alpha. Activities of PKC alpha and PKC zeta increased after exposure to asbestos, but neither isoform migrated to mitochondria. A general inhibitor of PKCs, bisindolylmaleimide I, had no effect on asbestos-induced apoptosis. Hydrogen peroxide (H2O2) induced activation of PKCs delta, alpha, zeta, and theta, translocation of PKC delta to mitochondria, and caspase-9 cleavage. However, H2O2-induced apoptosis was not inhibited by cell lines stably expressing either dnPKC delta or dnPKC alpha, suggesting that activation of PKC delta has a distinct role in the development of asbestos induced apoptosis. PMID- 12626343 TI - Peroxynitrite enhances interleukin-10 reduction in the release of neutrophil chemotactic activity. AB - Peroxynitrite, formed by nitric oxide and superoxide, has been shown to nitrate and reduce the function of proinflammatory proteins such as interleukin (IL)-8, monocyte chemoattractant protein-1, and eotaxin, but in contrast, to enhance the function of the anti-inflammatory cytokine IL-10 in reducing IL-1 release from blood monocytes. However, the effect of nitrated IL-10 on release of proinflammatory cytokines from lung epithelial cells is unknown. We hypothesized that peroxynitrite would enhance the capacity of human IL-10 to reduce inflammatory mediators released by epithelial cells. To test this hypothesis, recombinant human IL-10 was evaluated for its capacity to attenuate the release of neutrophil chemotactic activity and IL-8 from a human epithelial cell line in response to IL-1 beta and tumor necrosis factor-alpha. Neutrophil chemotactic activity and IL-8 in lung epithelial culture supernatant fluids were significantly lower after culture with nitrated human IL-10 compared with non nitrated human IL-10 controls (P < 0.05). Consistent with these results, nitrated human IL-10 attenuated IL-8 mRNA expression more than non-nitrated human IL-10 controls (P < 0.05). These data demonstrate that peroxynitrite exposed human IL 10 has enhanced anti-inflammatory activity and suggest that nitration may play a critical role in the regulation of inflammation within the lower respiratory tract. PMID- 12626344 TI - Expression of macrophage inflammatory protein-3 beta/CCL19 in pulmonary sarcoidosis. AB - In this study, messenger RNA (mRNA) expression for novel T lymphocyte chemoattractants, leukotactin-1, macrophage inflammatory protein (MIP)-3 alpha and MIP-3 beta was investigated in bronchoalveolar lavage fluid (BALF) cells from patients with sarcoidosis, a T cell-mediated disease with typical CD4+ lymphocyte alveolitis. Of these three chemokines, only MIP-3 beta mRNA was upregulated in sarcoidosis, and therefore, protein levels of this chemokine, its pharmacologic regulation, and association with disease clinical course were explored. MIP-3 beta protein concentrations were elevated in BALF from sarcoid patients compared with control subjects (p = 0.001) and in patients with chest X-ray stage II chemokine protein levels were increased compared with stage I (p = 0.003). MIP-3 beta protein was associated predominantly with alveolar macrophages and correlated with BALF lymphocytes and T cell subsets. mRNA expression for the MIP 3 beta receptor, CC chemokine receptor 7, was increased in sarcoidosis and correlated with MIP-3 beta protein levels. MIP-3 beta mRNA and protein expression in BALF cells was suppressed by dexamethasone and cyclosporine A in vitro. In conclusion, MIP-3 beta is implicated in T lymphocyte recruitment in sarcoidosis, is associated with disease progression, and is downregulated by drugs used for sarcoidosis treatment. This novel chemokine, therefore, represents a candidate for studies of sarcoidosis pathobiologic mechanisms. PMID- 12626346 TI - Early alterations in neutrophil activation are associated with outcome in acute lung injury. AB - Persistent elevations of proinflammatory cytokines in the lungs are associated with increased mortality from acute lung injury (ALI), suggesting that the degree of pulmonary inflammation is an important determinant of clinical course in ALI. The transcriptional regulatory factor nuclear factor-kappaB (NF-kappaB) is involved in modulating the expression of many cytokines and other proinflammatory mediators implicated in the development and progression of ALI. Because neutrophils appear to play a major role in the development of ALI, we examined the relationships between clinical outcome and activation of NF-kappaB in peripheral neutrophils from patients (n = 30) with sepsis-induced ALI. We found that nuclear translocation of NF-kappaB in this setting was dependent on the activation of p38 and Akt kinases. Diminished activation of NF-kappaB or Akt, but not p38, in the early postintubation period was associated with less time on the ventilator and improved survival in critically ill patients with ALI. These results suggest that early alterations in neutrophil activation patterns, particularly involving the ability to accumulate NF-kappaB to the nucleus after relevant stimuli, contribute to subsequent clinical course in ALI. PMID- 12626345 TI - Fibroblast growth factor receptor-1 and neonatal compensatory lung growth after exposure to 95% oxygen. AB - Neonatal rats exposed to 95% oxygen (O2) for 7 days from birth had inhibited lung growth, DNA synthesis, and secondary septation. These parameters were rapidly restored by a period of recovery in air. Northern and Western blot analysis and immunohistochemistry were used to screen for the fibroblast growth factor receptor-1 (FGF-R1) and its high affinity ligand, basic fibroblast growth factor (bFGF), which could have a role in this recovery process. Expression of bFGF in the lung was significantly reduced at the end of the 7-day exposure to 95% O2 and was increased after 3 days of recovery in air. Expression of FGF-R1 was not affected by exposure to 95% O2 or recovery in air. We hypothesized that the increase in bFGF after removal from 95% O2, acting through the FGF-R1, would be critical for compensatory growth. Intraperitoneal injection of soluble truncated FGF-R1 at the onset of the recovery phase arrested compensatory lung DNA synthesis and secondary septation seen in control animals after 3 days of recovery, confirming a role for FGF-R1 in this model of compensatory neonatal lung growth. PMID- 12626347 TI - Influence of Cheyne-Stokes respiration on cardiovascular oscillations in heart failure. AB - In patients with congestive heart failure, Cheyne-Stokes respiration is accompanied by oscillations in blood pressure and heart rate at a very low frequency. It is not known whether these cardiovascular oscillations are primarily related to oscillations in ventilation or oxyhemoglobin saturation. We hypothesized that abolition of the ventilatory oscillations of Cheyne-Stokes respiration by CO2 inhalation would eliminate accompanying oscillations in blood pressure and heart rate but that elimination of hypoxic dips by supplemental O2 would not. We studied 10 subjects with heart failure and Cheyne-Stokes respiration during sleep using frequency spectral analysis. During Cheyne-Stokes respiration, heart rate and blood pressure oscillated in association with respiratory oscillations at very low frequency. Inhalation of CO2 abolished Cheyne-Stokes respiration and associated oscillations in both blood pressure and heart rate. In contrast, inhalation of O2 sufficient to eliminate hypoxic dips had no significant effect on Cheyne-Stokes respiration, blood pressure (n = 6), or heart rate (n = 5). We conclude that ventilatory oscillations during Cheyne Stokes respiration rather than oscillations in oxygenation per se powerfully induce heart rate and blood pressure oscillations. Cheyne-Stokes respiration is therefore one of the mechanisms that contributes to the very low-frequency oscillations in heart rate and blood pressure observed in patients with heart failure. PMID- 12626348 TI - Beta 2 microglobulin knockout mice are resistant to lethal intraabdominal sepsis. AB - beta 2 microglobulin knockout (beta2M-/-) mice lack CD8+ T and natural killer T cells. We hypothesized that beta 2M-/- mice are resistant to lethal intraabdominal sepsis. To test this hypothesis, mortality, cytokine production, and physiologic function were assessed in beta 2M-/- mice during sepsis caused by cecal ligation and puncture (CLP). beta 2M-/- mice survived significantly longer than wild-type mice after CLP but ultimately exhibited 100% mortality. Treatment of beta 2M-/- mice with anti-asialoGM1 to deplete natural killer cells conferred greater than 70% long-term survival. Compared with wild-type mice, beta 2M-/- mice treated with anti-asialoGM1 produced decreased amounts of proinflammatory cytokines and did not exhibit hypothermia or metabolic acidosis after CLP. Adoptive transfer of CD8+ T and natural killer cells into beta 2M-/- mice treated with anti-asialoGM1 re-established CLP-induced mortality. CD8 knockout mice treated with anti-asialoGM1, which are specifically deficient in CD8+ T and natural killer cells, exhibited 40% long-term survival after CLP. Furthermore, treatment of wild-type mice with antibodies to CD8 and asialoGM1 conferred a significant survival benefit compared with wild-type mice treated with nonspecific IgG. These findings demonstrate that beta 2M-/- mice treated with anti-asialoGM1 are resistant to CLP-induced mortality and that depletion of CD8+ T and natural killer cells largely accounts for the survival benefit observed in these mice. PMID- 12626349 TI - Respiratory effects of gestational intermittent hypoxia in the developing rat. AB - Intermittent hypoxia (IH), one of the hallmarks of obstructive sleep apnea, occurs more frequently during pregnancy. We hypothesized that IH may lead to persistent postnatal changes in respiratory responses to acute hypoxia and may also lead to adverse effects on spatial function learning as revealed by the Morris water maze. To examine this issue, time-pregnant Sprague-Dawley rats were exposed to IH and room air (IHRA; 21 and 10% O2 alternations every 90 seconds) or to normoxia (RARA) until delivery. Ventilatory and metabolic responses to a 20 minute acute hypoxic challenge (10% O2) were conducted at postnatal ages 5, 10, 15, and 30 days. In addition, spatial task learning was assessed in the water maze at 1 and 4 months of age. Normoxic ventilation was higher at all time points in IHRA rats than in RARA rats (p < 0.01). Peak hypoxic ventilatory responses were attenuated in IHRA rats at 5 days of age and hypoxic ventilatory depression was accentuated at this age as well. However, ventilatory equivalents (minute ventilation/oxygen consumption) revealed significant reductions in peak hypoxic ventilatory responses of IHRA rats and hypoxic ventilatory depression at all postnatal ages (p < 0.01). Acquisition and retention of a spatial task were similar in the IHRA and RARA groups at both 1 and 4 months of age. We conclude that gestational intermittent hypoxia elicits long-lasting alterations in the control of breathing. We postulate that such IH-induced respiratory plasticity may create selective vulnerability to hypoxia during development. PMID- 12626350 TI - Arginine therapy: a new treatment for pulmonary hypertension in sickle cell disease? AB - Pulmonary hypertension is a life-threatening complication of sickle cell disease. L-Arginine is the nitrogen donor for synthesis of nitric oxide, a potent vasodilator that is deficient during times of sickle cell crisis. This deficiency may play a role in pulmonary hypertension. The enzyme arginase hydrolyzes arginine to ornithine and urea, and thus, it may compete with nitric oxide synthase, leading to decreased nitric oxide production. Nitric oxide therapy by inhalation has improved pulmonary hypertension associated with acute chest syndrome in sickle cell disease, and several studies demonstrate therapeutic benefits of arginine therapy for primary and secondary pulmonary hypertension. We sought to determine the effects of arginine therapy on pulmonary hypertension in patients with sickle cell disease. Arginase activity was also determined. Oral arginine produced a 15.2% mean reduction in estimated pulmonary artery systolic pressure (63.9 +/- 13 to 54.2 +/- 12 mm Hg, p = 0.002) after 5 days of therapy in 10 patients. Arginase activity was elevated almost twofold (p = 0.07) in patients with pulmonary hypertension and may limit arginine bioavailability. With limited treatment options and a high mortality rate for patients with sickle cell disease who develop pulmonary hypertension, arginine is a promising new therapy that warrants further investigation. PMID- 12626351 TI - Accentuated T helper type 2 airway response after allergen challenge in cyclooxygenase-1-/- but not cyclooxygenase-2-/- mice. AB - Acute pharmacologic inhibition of cyclooxygenase (COX)-1 or -2 during allergen sensitization and exposure leads to enhanced T helper type 2 (Th2) airway responses. COX-1 and -2 play functionally distinct roles in lymphocyte development, and consequently, genetic deficiency of either enzyme, as opposed to acute pharmacologic inhibition, may modulate Th2-mediated allergic airway disease differently. An ovalbumin-induced mouse model of allergic airway disease was used. The immunophenotype of bronchoalveolar lavage lymphocytes was assessed by flow cytometry, bronchoalveolar lavage cytokines, and chemokines were measured by enzyme-linked immunosorbent assay, adhesion molecule expression was assessed by immunoblotting in combination with immunohistochemistry, and bronchoconstriction was assessed by whole body plethysmography. The airways of COX-1-/- mice contained increased numbers of CD4+ and CD8+ T cells, exaggerated levels of the Th2 cytokines interleukin-4, -5, and -13, and increased levels of eotaxin and thymus- and activation-regulated chemokine. Allergen-induced bronchoconstriction was also increased in COX-1-/- mice. Vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 levels were increased in lungs of both COX-1-/- and COX-2-/- mice relative to wild type. These data suggest that genetic deficiency of COX-1 but not COX-2 modulates T cell recruitment, Th2 cytokine secretion, and lung function in the allergic airway. PMID- 12626352 TI - Neural expression and increased lavage fluid levels of secretoneurin in seasonal allergic rhinitis. AB - Secretoneurin is a neuropeptide potentially involved in migration of eosinophils, monocytes, and dendritic cells. Whether secretoneurin is present in the human airway mucosa and whether it is released at ongoing allergic airway inflammation is currently unknown. In patients with allergic rhinitis, we have explored the occurrence of secretoneurin in nasal mucosal biopsies and lavage fluids before and during natural allergen exposure. Immunohistochemical analysis revealed an abundance of nerves displaying secretoneurin immunoreactivity, which were distributed predominantly around blood vessels and submucosal glands. A majority of nerve fibers containing vesicular acetylcholine transporter, tyrosine hydroxylase, calcitonin gene-related peptide, and vasoactive intestinal peptide were also secretoneurin-immunoreactive, indicating a localization of secretoneurin in cholinergic, adrenergic, and sensory nerves. Lavage fluid levels of secretoneurin were increased at allergen exposure (p < 0.01-0.05). Levels of secretoneurin did not correlate with eosinophil cationic protein (rho = 0.1, p = 0.7). We conclude that secretoneurin has a widespread occurrence in nasal mucosal nerves of patients with seasonal allergic rhinitis and that increased nasal lavage fluid levels of secretoneurin may characterize ongoing allergen exposure. These data favor a role of secretoneurin in the local traffic of immune cells in human airway mucosa. PMID- 12626353 TI - The stromal derived factor-1/CXCL12-CXC chemokine receptor 4 biological axis in non-small cell lung cancer metastases. AB - Non-small cell lung cancer is characterized by a specific metastatic pattern. The mechanism for organ-specific metastasis is poorly understood, although evidence has suggested that the chemokine stromal derived factor-1 (CXCL12) and its cognate receptor CXCR4 may regulate breast cancer metastasis. We hypothesized that the CXCL12-CXCR4 biological axis is important in mediating non-small cell lung cancer metastases. Our results indicate that both non-small cell lung cancer tumor specimens resected from patients and non-small cell lung cancer cell lines express CXCR4, but not CXCL12. Non-small cell lung cancer cell lines undergo chemotaxis in response to CXCL12. CXCL12-CXCR4 activation of non-small cell lung cancer cell lines showed intracellular calcium mobilization and mitogen-activated protein kinase activation with enhanced extracellular signal-related kinase-1/2 phosphorylation without change in either proliferation or apoptosis. Target organs in a murine model that are the preferred destination of human non-small cell lung cancer metastases elaborate higher levels of CXCL12 than does the primary tumor, and suggest the generation of chemotactic gradients. The administration of specific neutralizing anti-CXCL12 antibodies to severe combined immunodeficient mice expressing human non-small cell lung cancer abrogated organ metastases, without affecting primary tumor-derived angiogenesis. These data suggest that the CXCL12-CXCR4 biological axis is involved in regulating the metastasis of non-small cell lung cancer. PMID- 12626354 TI - Fibroblast growth factors as regulators of central nervous system development and function. AB - Fibroblast growth factors (FGFs) are multifunctional signaling proteins that regulate developmental processes and adult physiology. Over the last few years, important progress has been made in understanding the function of FGFs in the embryonic and adult central nervous system. In this review, I will first discuss studies showing that FGF signaling is already required during formation of the neural plate. Next, I will describe how FGF signaling centers control growth and patterning of specific brain structures. Finally, I will focus on the function of FGF signaling in the adult brain and in regulating maintenance and repair of damaged neural tissues. PMID- 12626355 TI - Adiposity signals and food reward: expanding the CNS roles of insulin and leptin. AB - The hormones insulin and leptin have been proposed to act in the central nervous system (CNS) as adiposity signals as part of a theoretical negative feedback loop that senses the caloric stores of an animal and orchestrates adjustments in energy balance and food intake. Much research has provided support for both the existence of such a feedback loop and the specific roles that insulin and leptin may play. Most studies have focused on hypothalamic sites, which historically are implicated in the regulation of energy balance, and on the brain stem, which is a target for neural and humoral signals relating to ingestive acts. More recent lines of research, including studies from our lab, suggest that in addition to these CNS sites, brain reward circuitry may be a target for insulin and leptin action. These studies are reviewed together here with the goals of providing a historical overview of the findings that have substantiated the originally hypothesized negative feedback model and of opening up new lines of investigation that will build on these findings and allow further refinement of the model of adiposity signal/CNS feedback loop. The understanding of how motivational circuitry and its endocrine or neuroendocrine modulation contributes to normal energy balance regulation should expand possibilities for future therapeutic approaches to obesity and may lead to important insights into mental illnesses such as substance abuse or eating disorders. PMID- 12626356 TI - Role of oxidative stress in angiotensin-induced hypertension. AB - Infusion of ANG II at a rate not sufficient to evoke an immediate vasoconstrictor response, produces a slow increase in blood pressure. Circulating levels of ANG II may be within ranges found in normotensive individuals, although inappropriately high with respect to sodium intake. When ANG II levels are dissociated from sodium levels, oxidative stress (OXST) occurs, which can increase blood pressure by several mechanisms. These include inadequate production or reduction of bioavailability of nitric oxide, alterations in metabolism of arachidonic acid, resulting in an increase in vasoconstrictors and decrease in vasodilators, and upregulation of endothelin. This cascade of events appears to be linked, because ANG II hypertension can be blocked by inhibition of any factor located distally, blockade of ANG II, OXST, or endothelin. Such characteristics are shared by other models of hypertension, such as essential hypertension, hypertension induced by reduction in renal mass, and renovascular hypertension. Thus these findings are clinically important because they reveal 1) uncoupling between ANG II and sodium, which can trigger pathological conditions; 2) the various OXST mechanisms that may be involved in hypertension; and 3) therapeutic interventions for hypertension developed with the knowledge of the cascade involving OXST. PMID- 12626357 TI - Fever. PMID- 12626358 TI - Cardiovascular and renal sympathetic activation by blood-borne TNF-alpha in rat: the role of central prostaglandins. AB - In pathophysiological conditions, increased blood-borne TNF-alpha induces a broad range of biological effects, including activation of the hypothalamic-pituitary adrenal axis and sympathetic drive. In urethane-anesthetized adult Sprague-Dawley rats, we examined the mechanisms by which blood-borne TNF-alpha activates neurons in paraventricular nucleus (PVN) of hypothalamus and rostral ventrolateral medulla (RVLM), two critical brain regions regulating sympathetic drive in normal and pathophysiological conditions. TNF-alpha (0.5 microg/kg), administered intravenously or into ipsilateral carotid artery (ICA), activated PVN and RLVM neurons and increased sympathetic nerve activity, arterial pressure, and heart rate. Responses to intravenous TNF-alpha were not affected by vagotomy but were reduced by mid-collicular decerebration. Responses to ICA TNF-alpha were substantially reduced by injection of the cyclooxygenase inhibitor ketorolac (150 microg) into lateral ventricle. Injection of PGE(2) (50 ng) into lateral ventricle or directly into PVN increased PVN or RVLM activity, respectively, and sympathetic drive, with shorter onset latency than blood-borne TNF-alpha. These findings suggest that blood-borne cytokines stimulate cardiovascular and renal sympathetic responses via a prostaglandin-dependent mechanism operating at the hypothalamic level. PMID- 12626359 TI - Increased platelet-activating factor-induced periventricular brain microvascular constriction associated with immaturity. AB - Oxidant stress contributes to the pathogenesis of hypoxic-ischemic encephalopathies. Platelet-activating factor (PAF) is generated during oxidant stress. We studied the vasomotor mode of actions of PAF on periventricular (PV) microvessels of fetal ( approximately 75% of term), newborn (1-3 days), and adult pigs. PAF constricted PV microvessels from fetal (29.27 +/- 2.6%) and newborn (22.14 +/- 3.2%) pigs but was ineffective in adults (<2.5%). Specific [(3)H]PAF binding was greater in fetus and newborn than in adults; a concordant developmental PAF-induced inositol phosphate formation was observed. PAF-induced vasoconstriction was abrogated by thromboxane A(2) (TXA(2)) synthase and receptor inhibitors, calcium channel blockers, and by removal of endothelium; vasoconstriction to TXA(2) mimetic U-46619 did not differ with age. Immunoreactive TXA(2) synthase expression and PAF-evoked TXA(2) formation revealed a fetus> newborn>adult profile. Thus the greater PAF-induced PV microvascular constriction in younger subjects seems attributable to greater PAF receptor density and mostly secondary to TXA(2) formation from endothelium. The resulting decrease in blood flow may contribute to the increased vulnerability of the PV brain regions to oxidant stress-induced injury in immature subjects. PMID- 12626360 TI - 5-Aminoimidazole-4-carboxamide 1-beta -D-ribofuranoside (AICAR) stimulates myocardial glycogenolysis by allosteric mechanisms. AB - We tested the hypothesis that activation of AMP-activated protein kinase (AMPK) promotes myocardial glycogenolysis by decreasing glycogen synthase (GS) and/or increasing glycogen phosphorylase (GP) activities. Isolated working hearts from halothane-anesthetized male Sprague-Dawley rats perfused in the absence or presence of 0.8 or 1.2 mM 5-aminoimidazole-4-carboxamide 1-beta-d-ribofuranoside (AICAR), an adenosine analog and cell-permeable activator of AMPK, were studied. Glycogen degradation was increased by AICAR, while glycogen synthesis was not affected. AICAR increased myocardial 5-aminoimidazole-4-carboxamide 1-beta-d ribofuranotide (ZMP), the active intracellular form of AICAR, but did not alter the activity of GS and GP measured in tissue homogenates or the content of glucose-6-phosphate and adenine nucleotides in freeze-clamped tissue. Importantly, the calculated intracellular concentration of ZMP achieved in this study was similar to the K(m) value of ZMP for GP determined in homogenates of myocardial tissue. We conclude that the data are consistent with allosteric activation of GP by ZMP being responsible for the glycogenolysis caused by AICAR in the intact rat heart. PMID- 12626361 TI - Fos, RVLM-projecting neurons, and spinally projecting neurons in the PVN following hypertonic saline infusion. AB - Hypertonic saline (HTS; 1.7 M) infused intravenously into conscious rats increases the production of Fos, a marker of cell activation, in the hypothalamic paraventricular nucleus (PVN). The parvocellular PVN contains subpopulations of neurons. However, which subpopulations are activated by HTS is unknown. We determined whether PVN neurons that innervate the rostral ventrolateral medulla (RVLM) or the spinal cord (important autonomic sites) expressed Fos following HTS. Experiments were performed 24-96 h after chronic implantation of an intravenous cannula. HTS significantly increased the number of Fos-positive cells. In the parvocellular PVN, the maximum number of Fos-positive cells occurred rostral of the anterior-posterior level at which the number of neurons that projected to the medulla or spinal cord peaked. Compared with controls, HTS did not significantly increase the number of double-labeled neurons. These findings demonstrate that an elevation in plasma osmolality activates PVN neurons but not the subgroups of PVN neurons with projections to the RVLM or to the spinal cord. PMID- 12626362 TI - Interaction of prostaglandins with the renin-angiotensin system. PMID- 12626363 TI - PTHrP regulates cerebral blood flow and is neuroprotective. PMID- 12626364 TI - Adenovirus-mediated human prostasin gene delivery is linked to increased aldosterone production and hypertension in rats. AB - Prostasin has been demonstrated to be an activator of epithelial sodium channels in cultured renal and bronchial epithelial cells. In this study, we evaluated the effects of adenovirus-mediated gene transfer of human prostasin on blood pressure regulation and sodium reabsorption in Wistar rats. Expression of human prostasin mRNA was identified in rat adrenal gland, liver, kidney, heart, lung, and aorta, and immunoreactive human prostasin was detected in the circulation and urine of rats receiving prostasin gene transfer. A single injection of adenovirus carrying the prostasin gene caused prolonged increases in blood pressure for 3-4 wk. Blood pressure increase was accompanied by elevated plasma aldosterone levels and reduced plasma renin activity. The increase in blood pressure and plasma aldosterone levels as well as the reduction of plasma renin activity correlated with the expression of human prostasin transgene. Elevated plasma aldosterone levels were detected at 3 days after gene transfer before the development of hypertension, indicating that stimulation of mineralocorticoid production is the primary target of prostasin. Prostasin gene transfer significantly reduced urinary K(+) excretion but increased urinary Na(+) and kallikrein excretion. Elevated renal kallikrein levels promote natriuresis, which may lead to sodium escape and prevent further increases of blood pressure after prostasin gene transfer. In summary, these results suggest that prostasin participates in blood pressure and electrolyte homeostasis by regulating the renin-angiotensin aldosterone and kallikrein-kinin systems. PMID- 12626365 TI - Mechanisms regulating the marked seasonal variation in melatonin synthesis in the European hamster pineal gland. AB - Like many wild species, the European hamster (Cricetus cricetus) adapts to the marked seasonal changes in its environment, namely by hibernation and inhibition of sexual activity in winter. These annual functions are driven by the variation in the environmental factors (light, temperature) that are transmitted to the body through large variations in the duration and amplitude of the nocturnal melatonin rhythm. Here we report that the seasonal variation in melatonin synthesis is mainly driven by arylalkylamine N-acetyltransferase gene transcription and enzyme activation. This, however, does not exclude participation of hydroxyindole-O-methyltransferase, which may relay environmental temperature information. The in vivo experiments show that norepinephrine stimulates melatonin synthesis, this effect being gated at night. The possibility that the variation in pineal metabolism depends on a seasonal change in the suprachiasmatic nuclei clock circadian activity that is transmitted by norepinephrine is discussed. PMID- 12626366 TI - Androgen-induced activation of 20-HETE production may contribute to gender differences in blood pressure regulation. PMID- 12626367 TI - Length-dependent regulation of basal myosin phosphorylation and force in detrusor smooth muscle. AB - Urinary bladder (detrusor) smooth muscle is active in the absence of an external stimulus. Tone occurs even "at rest" during the filling phase, and it is elevated in patients with overactive bladder. This study examined the role of muscle length on tone and the level of basal myosin light chain phosphorylation (MLC(20P)). MLC(20P) was 23.9 +/- 1% (n = 58) at short lengths (zero preload; L(z)). An increase in length from L(z) to the optimal length for contraction (L(o)) caused a reduction in MLC(20P) to 15.8 +/- 1% (n = 49). Whereas 10 microM staurosporine reduced MLC(20P) at L(z), 1 microM staurosporine, a Ca(2+)-free solution, and inhibitors of MLC kinase, protein kinase C (PKC) and RhoA kinase (ROK) did not. However, 1 microM staurosporine and inhibitors of ROK inhibited MLC(20P) and tone at L(o). These data support the hypothesis that a Ca(2+) independent kinase, possibly ZIP-like kinase, regulates MLC(20P) at L(z), whereas in detrusor stretched to L(o), additional kinases, such as ROK, participate. PMID- 12626368 TI - Sympathoinhibitory pathway from caudal midline medulla to RVLM is independent of baroreceptor reflex pathway. AB - Glutamate stimulation of the caudal midline medulla (CMM) causes profound sympathoinhibition due to GABAergic inhibition of presympathetic neurons in the rostral ventrolateral medulla (RVLM). We investigated whether the sympathoinhibitory pathway from CMM to RVLM, like the central baroreceptor reflex pathway, includes a glutamatergic synapse in the caudal ventrolateral medulla (CVLM). In pentobarbital sodium-anesthetized rats, the RVLM on one side was inhibited by a muscimol microinjection. Then the response evoked by glutamate microinjections into the CMM or by baroreceptor stimulation was determined before and after 1) microinjection of the GABA receptor antagonist bicuculline into the RVLM on the other side or 2) microinjections of the glutamate receptor antagonist kynurenate bilaterally into the CVLM. Bicuculline in the RVLM greatly reduced both CMM- and baroreceptor-evoked sympathoinhibition. Compared with the effect of vehicle solution, kynurenate in the CVLM greatly reduced baroreceptor-evoked sympathoinhibition, whereas its effect on CMM-evoked sympathoinhibition was not different from that of the vehicle solution. These findings indicate that the output pathway from CMM sympathoinhibitory neurons, unlike the baroreceptor and other reflex sympathoinhibitory pathways, does not include a glutamatergic synapse in the CVLM. PMID- 12626369 TI - ANG II is involved in the LPS-induced production of proinflammatory cytokines in dehydrated rats. AB - We have previously reported results that led us to speculate that ANG II is involved in the LPS-induced production of proinflammatory cytokines, especially under dehydrated conditions. To test this possibility, in this study we examined the effects of an angiotensin-converting enzyme (ACE) inhibitor and an antagonist of the type-1 ANG II receptor (AT(1) receptor) on the LPS-induced production of the proinflammatory cytokines IL-1 and IL-6 in dehydrated rats. A single intravenous injection of LPS induced a marked increase in the expression of IL 1beta mRNA in the liver, an effect that was significantly attenuated by pretreatment with the ACE inhibitor. Furthermore, the ACE inhibitor reduced the LPS-induced increase in the hepatic concentration of IL-1beta protein. When the AT(1)-receptor antagonist was given intravenously before the LPS, the increase in the hepatic concentration of IL-1beta was significantly reduced. Finally, the ACE inhibitor reduced the LPS-induced increase in the plasma concentration of IL-6. These results represent the first in vivo evidence that ANG II and its AT(1) receptor play important roles in the production of proinflammatory cytokines that is induced by LPS under dehydrated conditions. PMID- 12626370 TI - Chronic angiotensin II infusion attenuates the renal sympathoinhibitory response to acute volume expansion. AB - In this study the hypothesis was tested that chronic infusion of ANG II attenuates acute volume expansion (VE)-induced inhibition of renal sympathetic nerve activity (SNA). Rats received intravenous infusion of either vehicle or ANG II (12 ng. kg(-1). min(-1)) for 7 days. ANG II-infused animals displayed an increased contribution of SNA to the maintenance of mean arterial pressure (MAP) as indicated by ganglionic blockade, which produced a significantly (P < 0.01) greater decrease in MAP (75 +/- 3 mmHg) than was observed in vehicle-infused (47 +/- 8 mmHg) controls. Rats were then anesthetized, and changes in MAP, mean right atrial pressure (MRAP), heart rate (HR), and renal SNA were recorded in response to right atrial infusion of isotonic saline (20% estimated blood volume in 5 min). Baseline MAP, HR, and hematocrit were not different between groups. Likewise, MAP was unchanged by acute VE in vehicle-infused animals, whereas VE induced a significant bradycardia (P < 0.05) and increase in MRAP (P < 0.05). MAP, MRAP, and HR responses to VE were not statistically different between animals infused with vehicle vs. ANG II. In contrast, VE significantly (P < 0.001) reduced renal SNA by 33.5 +/- 8% in vehicle-infused animals but was without effect on renal SNA in those infused chronically with ANG II. Acutely administered losartan (3 mg/kg iv) restored VE-induced inhibition of renal SNA (P < 0.001) in rats chronically infused with ANG II. In contrast, this treatment had no effect in the vehicle-infused group. Therefore, it appears that chronic infusion of ANG II can attenuate VE-induced renal sympathoinhibition through a mechanism requiring AT(1) receptor activation. The attenuated sympathoinhibitory response to VE in ANG II-infused animals remained after arterial barodenervation and systemic vasopressin V(1) receptor antagonism and appeared to depend on ANG II being chronically increased because ANG II given acutely had no effect on VE induced renal sympathoinhibition. PMID- 12626371 TI - The embryo makes red blood cell progenitors in every tissue simultaneously with blood vessel morphogenesis. AB - During embryonic life, hematopoiesis occurs first in the yolk sac, followed by the aorto-gonado-mesonephric region, the fetal liver, and the bone marrow. The possibility of hematopoiesis in other embryonic sites has been suspected for a long time. With the use of different methodologies (transgenic mice, electron microscopy, laser capture microdissection, organ culture, and cross-transplant experiments), we show that multiple regions within the embryo are capable of forming blood before and during organogenesis. This widespread phenomenon occurs by hemo-vasculogenesis, the formation of blood vessels accompanied by the simultaneous generation of red blood cells. Erythroblasts develop within aggregates of endothelial cell precursors. When the lumen forms, the erythroblasts "bud" from endothelial cells into the forming vessel. The extensive hematopoietic capacity found in the embryo helps explain why, under pathological circumstances such as severe anemia, extramedullary hematopoiesis can occur in any adult tissue. Understanding the intrinsic ability of tissues to manufacture their own blood cells and vessels has the potential to advance the fields of organogenesis, regeneration, and tissue engineering. PMID- 12626372 TI - An evolutionary approach for identifying potential transcription factor binding sites: the renin gene as an example. AB - Evolutionary pressure has resulted in the conservation of certain nucleotide sequences. These conserved regions are potentially important for certain functions. Here we give an example of a comparison between noncoding sequences combined with other independent database information to shed light onto the regulation of the renin gene, a gene that has great importance for cardiovascular and renal homeostasis. To combine the information regarding conservation and weight matrices of transcription factor (TF) binding sites, an algorithm was developed (TFprofile). Notably, a local peak in the resulting binding profile coincides with a previously experimentally identified regulatory region for the renin gene. The existence of further peaks in the binding profile in the conserved 3.9-kb-long hRENc DNA block upstream of the renin gene suggests additional regions of potential importance for gene regulation. The algorithm TFprofile may be used to integrate information on cross-species evolutionary conservation and aspects of TF binding characteristics to provide putative regulatory DNA regions for experimental verification. PMID- 12626373 TI - Determinants of success of inpatient clinical information systems: a literature review. AB - We reviewed the English and Dutch literature on evaluations of patient care information systems that require data entry by health care professionals published from 1991 to 2001. Our objectives were to identify attributes that were used to assess the success of such systems and to test the ability of a framework developed by Delone and McLean for management information systems(1) to categorize these attributes correctly. The framework includes six dimensions or success factors: system quality, information quality, usage, user satisfaction, individual impact, and organizational impact. Thirty-three papers were selected for complete review. Types of study design included descriptive, correlational, comparative, and case studies. A variety of relevant attributes could be assigned to the six dimensions in the Delone and McLean framework, but some attributes, predominantly in cases of failure, did not fit any of the categories. They related to contingent factors, such as organizational culture. Our review points out the need for more thorough evaluations of patient care information systems that look at a wide range of factors that can affect the relative success or failure of these systems. PMID- 12626375 TI - Building and evaluation of a structured representation of pharmacokinetics information presented in SPCs: from existing conceptual views of pharmacokinetics associated with natural language processing to object-oriented design. AB - OBJECTIVE: Develop a detailed representation of pharmacokinetics (PK), derived from the information in Summaries of Product Characteristics (SPCs), for use in computerized systems to help practitioners in pharmaco-therapeutic reasoning. METHODS: Available knowledge about PK was studied to identify main PK concepts and organize them in a preliminary generic model. The information from 1950 PK SPC-texts in the French language was studied using a morpho-syntactic analyzer. It produced a list of candidate terms (CTs) from which those describing main PK concepts were selected. The contexts in which they occurred were explored to discover co-occurring CTs. The regrouping according to CT semantic types led to a detailed object-oriented model of PK. The model was evaluated. A random sample of 100 PK texts structured according to the model was judged for completeness and semantic accuracy by 8 experts who were blinded to other experts' responses. RESULTS: The PK text file contained about 300000 words, and the morpho-syntactic analysis extracted 17520 different CTs. The context of 592 CTs was studied and used to deduce the PK model. It consists of four entities: the information about the real PK process, the experimental protocol, the mathematical modeling, and the influence of factors causing variation. Experts judged that the PK model represented the information in 100 sample PK texts completely in 89% of cases and nearly completely in the other 11%. There was no distortion of meaning in 98% of cases and little distortion in the remainder. CONCLUSION: The PK model seems to be applicable to all SPCs and can be used to retranscribe legal information from PK sections of SPCs into structured databases. PMID- 12626374 TI - Generating hypotheses by discovering implicit associations in the literature: a case report of a search for new potential therapeutic uses for thalidomide. AB - The availability of scientific bibliographies through online databases provides a rich source of information for scientists to support their research. However, the risk of this pervasive availability is that an individual researcher may fail to find relevant information that is outside the direct scope of interest. Following Swanson's ABC model of disjoint but complementary structures in the biomedical literature, we have developed a discovery support tool to systematically analyze the scientific literature in order to generate novel and plausible hypotheses. In this case report, we employ the system to find potentially new target diseases for the drug thalidomide. We find solid bibliographic evidence suggesting that thalidomide might be useful for treating acute pancreatitis, chronic hepatitis C, Helicobacter pylori-induced gastritis, and myasthenia gravis. However, experimental and clinical evaluation is needed to validate these hypotheses and to assess the trade-off between therapeutic benefits and toxicities. PMID- 12626376 TI - A consensus statement on considerations for a successful CPOE implementation. AB - In May of 2001, thirteen experts on computerized provider order entry (CPOE) from around the world gathered at a 2-day conference to develop a consensus statement on successful CPOE implementation. A qualitative research approach was used to generate and validate a list of categories and considerations to guide CPOE implementation. PMID- 12626378 TI - Web messaging: a new tool for patient-physician communication. AB - OBJECTIVE: There is a high demand by patients to communicate electronically with their doctor. This study evaluates the use of a web messaging system by staff and patients of UC Davis community Primary Care Network (PCN) clinic. DESIGN: Eight providers and their staff and patients were surveyed on use of this web messaging system, and physician productivity was measured with Relative Value Unit (RVU) and office visit data. RESULTS: 36.9% (238/645) of registered users responded to the survey. The web messaging system was preferred over phone calls by both providers and patients for the communication of non-urgent problems. A great majority of patients found it easy to use (88.8% or 206/232) and were satisfied (85.8% or 199/232). Satisfaction was significantly associated with timely provider response (Goodman-Kruskal Gamma = 0.667, 95% CI = 0.546-0.789). Clinicians were also favorable to the system and, despite concerns, were not inundated with messages. Most found it easy to use, perceived it to improve patient communication, and valued the insurance reimbursement capability. Furthermore, the system did not have a negative impact on physician productivity. CONCLUSION: A patient-provider web messaging system, which provides a combination of security and access controls, customized routing, rich knowledge content, and insurance reimbursement capability, is a useful addition to the array of communication options available to health care providers and their patients. PMID- 12626379 TI - Clinical team functioning and IT innovation: a study of the diffusion of a point of-care online evidence system. AB - OBJECTIVES: To investigate the association between clinical team functioning and diffusion (awareness, use, and impact) of a 24-hour online evidence retrieval system. To examine the relationships between clinical team characteristics and the adoption of the online evidence system. DESIGN: 18 clinical teams, consisting of 180 clinicians from three Australian hospitals, were identified and studied. Teams were categorized as small ( 15). MEASUREMENTS: Clinical team functioning was assessed using the Team Climate Inventory (TCI). Awareness, use, and impact of an online evidence retrieval system were measured using a self-administered questionnaire. The relationships between TCI scores and awareness, use, and impact were examined using t-tests and one-way ANOVAs. Chi square analyses were used to examine differences between small and large teams. RESULTS were interpreted within a diffusion of innovations framework. RESULTS: Clinical team functioning was not related to awareness or use of the online evidence retrieval system. However, clinical team functioning was significantly associated with the impact of online evidence in terms of reported experience of improved patient care following system use. Clinicians in small teams ( 15) teams. CONCLUSIONS: Team functioning had the greatest impact on the fourth stage of innovation diffusion, the effective use of online evidence for clinical care. This supports Rogers' diffusion of innovation theory, to the effect that different types of communication about an innovation are important at different stages in the diffusion process. Members of small teams were more aware of the system than members of large teams. Team functioning is amenable to improvement through interventions. The findings suggest that the role of team climate in the diffusion of information systems is a promising area for future research. PMID- 12626381 TI - Predicting the molecular shape of polysaccharides from dynamic interactions with water. AB - How simple monosaccharides, once polymerized, become the basis for structural materials remains a mystery. A framework is developed to investigate the role of water in the emergence of dynamic structure in polysaccharides, using the important beta(1-->4) linkage as an example. This linkage is studied within decasaccharide fragments of cellulose, chitin, mannan, xylan, and hyaluronan, using molecular simulations in the presence of explicit water solvent. Although cellulose, mannan, chitin, and xylan are chemically similar, their intramolecular hydrogen-bond dynamics and interaction with water are predicted to differ. Cellulose, mannan, and chitin favor relatively static intramolecular hydrogen bonds, xylan prefers dynamic water bridges, and multiple water configurations are predicted at the beta(1-->4) linkages of hyaluronan. With such a variety of predicted dynamics, the hypothesis that the beta(1-->4) linkage is stabilized by intramolecular hydrogen bonds was rejected. Instead, it is proposed that favored molecular configurations are consistent with maximum rotamer and water degrees of freedom, explaining observations made previously by X-ray diffraction. Furthermore, polysaccharides predicted to be conformationally restricted in simulations (cellulose, chitin, and mannan) prefer the solid state in reality, even as oligosaccharides. Those predicted to be more flexible (xylan and hyaluronan) are known to be soluble, even as high polymers. Therefore an intriguing correlation between chemical composition, water organization, polymer properties, and biological function is proposed. PMID- 12626380 TI - Evaluating the impact of information technology on medication errors: a simulation. PMID- 12626382 TI - Chemoenzymatic synthesis and application of glycopolymers containing multivalent sialyloligosaccharides with a poly(L-glutamic acid) backbone for inhibition of infection by influenza viruses. AB - Highly water-soluble glycopolymers with poly(alpha-L-glutamic acid) (PGA) backbones carrying multivalent sialyl oligosaccharides units were chemoenzymatically synthesized as polymeric inhibitors of infection by human influenza viruses. p-Aminophenyl disaccharide glycosides were coupled with gamma carboxyl groups of PGA side chains and enzymatically converted to Neu5Acalpha2 3Galbeta1-4GlcNAcbeta-, Neu5Acalpha2-6Galbeta1-4GlcNAcbeta-, Neu5Acalpha2 3Galbeta1-3GalNAcalpha-, and Neu5Acalpha2-3Galbeta1-3GalNAcbeta- units, respectively, by alpha2,3- or alpha2,6-sialytransferases. The glycopolymers synthesized were used for neutralization of human influenza A and B virus infection as assessed by measurement of the degree of cytopathic inhibitory effect in virus-infected MDCK cells. Among the glycopolymers tested, alpha2,6 sialo-PGA with a high molecular weight (260 kDa) most significantly inhibited infection by an influenza A virus, strain A/Memphis/1/71 (H3N2), which predominantly binds to alpha2-6 Neu5Ac residue. The alpha2,6-sialo-PGA also inhibited infection by an influenza B virus, B/Lee/40. The binding preference of viruses to terminal sialic acids was affected by core determinants of the sugar chain, Galbeta1-4GlcNAcbeta- or Galbeta1-3GalNAcalpha/beta- units. Inhibition of infection by viruses was remarkably enhanced by increasing the molecular weight and sialic acid content of glycopolymers. PMID- 12626383 TI - GALT deficiency causes UDP-hexose deficit in human galactosemic cells. AB - Previously we reported that stable transfection of human UDP-glucose pyrophosphorylase (hUGP2) rescued galactose-1-phosphate uridyltransferase (GALT) deficient yeast from "galactose toxicity." Here we test in human cell lines the hypothesis that galactose toxicity was caused by excess accumulation of galactose 1-phosphate (Gal-1-P), inhibition of hUGP2, and UDP-hexose deficiency. We found that SV40-transformed fibroblasts derived from a galactosemic patient accumulated Gal-1-P from 1.2+/-0.4 to 5.2+/-0.5 mM and stopped growing when transferred from 0.1% glucose to 0.1% galactose. Control fibroblasts accumulated little Gal-1-P and continued to grow. The GALT-deficient cells had 157+/-10 micromoles UDP glucose/100 g protein and 25+/-5 micromoles UDP-galactose/100 g protein when grown in 0.1% glucose. The control cells had 236+/-25 micromoles UDP- glucose/100 g protein and 82+/-10 micromoles UDP-galactose/100 g protein when grown in identical medium. When we transfected the GALT-deficient cells with either the hUGP2 or GALT gene, their UDP-glucose content increased to 305+/-28 micromoles/100 g protein (hUGP2-transfected) and 210+/-13 micromoles/100 g protein (GALT-transfected), respectively. Similarly, UDP-galactose content increased to 75+/-12 micromoles/100 g protein (hUGP2-transfected) and 55+/-9 micromoles/100 g protein (GALT-transfected), respectively. Though the GALT transfected cells grew in 0.1% galactose with little accumulation of Gal-1-P (0.2+/-0.02 mM), the hUGP2-transfected cells grew but accumulated some Gal-1-P (3.1+/-0.4 mM). We found that 2.5 mM Gal-1-P increased the apparent KM of purified hUGP2 for glucose-1-phosphate from 19.7 microM to 169 microM, without changes in apparent Vmax. The Ki of the reaction was 0.47 mM. Gal-1-P also inhibited UDP-N-acetylglucosamine pyrophosphorylase, which catalyzes the formation of UDP-N-acetylglucosamine. We conclude that intracellular concentrations of Gal-1-P found in classic galactosemia inhibit UDP-hexose pyrophosphorylases and reduce the intracellular concentrations of UDP-hexoses. Reduced Sambucus nigra agglutinin binding to glycoproteins isolated from cells with increased Gal-1-P is consistent with the resultant inhibition of glycoprotein glycosylation. PMID- 12626384 TI - A biochemical and pharmacological comparison of enzyme replacement therapies for the glycolipid storage disorder Fabry disease. AB - Fabry disease is a lysosomal storage disease arising from deficiency of the enzyme alpha-galactosidase A. Two recombinant protein therapeutics, Fabrazyme (agalsidase beta) and Replagal (agalsidase alfa), have been approved in Europe as enzyme replacement therapies for Fabry disease. Both contain the same human enzyme, alpha-galactosidase A, but they are produced using different protein expression systems and have been approved for administration at different doses. To determine if there is recognizable biochemical basis for the different doses, we performed a comparison of the two drugs, focusing on factors that are likely to influence biological activity and availability. The two drugs have similar glycosylation, both in the type and location of the oligosaccharide structures present. Differences in glycosylation were mainly limited to the levels of sialic acid and mannose-6-phosphate present, with Fabrazyme having a higher percentage of fully sialylated oligosaccharides and a higher level of phosphorylation. The higher levels of phosphorylated oligomannose residues correlated with increased binding to mannose-6-phosphate receptors and uptake into Fabry fibroblasts in vitro. Biodistribution studies in a mouse model of Fabry disease showed similar organ uptake. Likewise, antigenicity studies using antisera from Fabry patients demonstrated that both drugs were indistinguishable in terms of antibody cross reactivity. Based on these studies and present knowledge regarding the influence of glycosylation on protein biodistribution and cellular uptake, the two protein preparations appear to be functionally indistinguishable. Therefore, the data from these studies provide no rationale for the use of these proteins at different therapeutic doses. PMID- 12626385 TI - Cloning and functional expression of a novel GDP-6-deoxy-D-talose synthetase from Actinobacillus actinomycetemcomitans. AB - Actinobacillus actinomycetemcomitans is a Gram-negative coccobacillus that can cause various forms of severe periodontitis and other nonoral infections in human patients. The serotype a-specific polysaccharide antigen of A. actinomycetemcomitans contains solely 6-deoxy-D-talose and its O-2 acetylated modification. This polysaccharide is synthesized from the donor GDP-6-deoxy-D talose with the relevant talosylation enzyme(s). In the synthesis of GDP-6- deoxy D-talose, GDP-D-mannose is first converted by GDP-mannose-4,6-dehydratase (GMD) to GDP-4-keto-6-deoxy-D-mannose and then reduced to GDP-6-deoxy-D-talose by GDP-6 deoxy-D-talose synthetase (GTS). In this study, we cloned and overexpressed in Escherichia coli the A. actinomycetemcomitans GTS enzyme responsible for the synthesis of GDP-6-deoxy-D-talose. The recombinant A. actinomycetemcomitans GTS enzyme expressed in E. coli converted the GDP-4-keto-6-deoxy-intermediate to a novel GDP-deoxyhexose. The synthesized GDP-deoxyhexose was shown to be GDP-6 deoxy-D-talose by HPLC, MALDI-TOF MS, and NMR spectroscopy. The functional expression of gts provides another enzymatically defined pathway for the synthesis of GDP-deoxyhexoses, which can be used as donors for the corresponding glycosyltransferases. PMID- 12626386 TI - Specificity of human anti-NOR antibodies, a distinct species of "natural" anti alpha-galactosyl antibodies. AB - Natural anti-NOR antibodies are common in human sera and agglutinate human erythrocytes of a rare NOR phenotype. The NOR phenotype-related antigens are unique neutral glycosphingolipids recognized by these antibodies and Griffonia simplicifolia IB4 isolectin (GSL-IB4). The oligosaccharide chains of NOR glycolipids are terminated by Galalpha1-4GalNAcbeta1-3Galalpha units. To characterize the specificity of anti-NOR antibodies and compare it with specificities of GSL-IB4 and known anti-Galalpha1,3Gal antibodies, alpha galactosylated saccharides and saccharide-polyacrylamide conjugates were used. New synthetic oligosaccharides, corresponding to the terminal di- and trisaccharide sequence of NOR glycolipids and the conjugate of the NOR-tri with HSA were included. These compounds were tested by microtiter plate ELISA and hemagglutination inhibition. Anti-NOR antibodies reacted most strongly with Galalpha1-4GalNAcbeta1-3Gal (NOR-tri), and over 100 times less strongly with Galalpha1-4GalNAc (NOR-di). The antibodies reacted also with Galalpha1-4Gal and Galalpha1-4Galbeta1-4GlcNAc, similarly as with NOR-di but not with other tested compounds. In turn, anti-Galalpha1,3Gal antibodies reacted most strongly with Galalpha1-3Gal and were very weakly inhibited by the NOR-related oligosaccharides (weaker than by galactose), and NOR-tri was less active than NOR-di. GSL-IB4 reacted with all tested alpha-galactosylated saccharides and conjugates, including the similarly active NOR-tri and NOR-di. These results showed that anti NOR represent a new species of anti-alpha-galactosyl antibodies with high affinity for the Galalpha1-4GalNAcbeta1-3Gal sequence present in rare NOR erythrocytes. PMID- 12626387 TI - Distinct localization of MUC5B glycoforms in the human salivary glands. AB - Salivary mucins, encoded by the MUC5B gene, make up a heterogeneous family of molecules, which are secreted by several glands, including the submandibular, sublingual, and palatine glands. Previous studies have shown that heterogeneity in the salivary mucin population is related to its multiglandular origin. In the present study we address the question to what extent the mucin (MUC5B) population from a single human salivary gland is made up of different glycoforms. Using monoclonal antibodies to defined protein and sulfated carbohydrate epitopes specific to MUC5B, we conduct an immunohistochemical study of different salivary gland types, including submandibular, sublingual, and labial glands. In all tissues studied we found a mosaic expression pattern of sulfo-Lewis a antigen, recognized by mAb F2, which in salivary glands is exclusively present on MUC5B. On the other hand, mucous acini were uniformly labeled by mAb EU-MUC5Bb, evoked against a peptide-stretch of the tandem repeat region of MUC5B. Double staining with both antibodies confirmed the presence of MUC5B-positive/sulfo-Lewis a positive cells, as well as MUC5B-positive/sulfo-Lewis a-negative cells within one glandular unit. These results indicate that one and the same salivary gland synthesizes different MUC5B glycoforms. PMID- 12626388 TI - Multiple transcription initiation and alternative splicing in the 5' untranslated region of the core 2 beta1-6 N-acetylglucosaminyltransferase I gene. AB - The glycosyltransferase core 2 beta1,6 N-acetylglucosaminyltransferase I (C2GnT I) plays an important regulatory role in the synthesis of biologically significant oligosaccharide structures. This gene is expressed in a variety of cell types, including lymphocytes and mucin-producing cells. The expression pattern of this gene suggests a complex system of regulation. To investigate the molecular regulation of this gene and locate potential promoter elements, rapid amplification of cDNA ends (RACE) analysis was used to determine the 5' ends of the C2GnT I mRNAs from a number of tissues. These experiments identified five C2GnT I mRNAs that are different in their 5' untranslated regions. The RACE cDNAs had four different 5' terminal sequences (exons A, B, D, and E'), suggesting four transcription initiation sites. One mRNA form was the result of alternative exon (exon C) utilization. These exons are spread across 60 kb of DNA on human chromosome 9, and all splice to the exon (exon F) that contains the C2GnT I coding region. Reverse transcription polymerase chain reaction experiments using primers specific for each of the four 5' end exon sequences revealed that the 5' terminal exons are differentially expressed, suggesting tissue specificity for the different 5' untranslated regions. These findings are consistent with the presence of multiple tissue-specific promoters for the C2GnT I gene. PMID- 12626389 TI - Increased fucosylation and reduced branching of serum glycoprotein N-glycans in all known subtypes of congenital disorder of glycosylation I. AB - The N-glycans present on the total mixture of serum glycoproteins (serum N glycome) were analyzed in 24 subjects with congenital disorder of glycosylation type I (CDG-I) and 7 healthy, age-matched individuals. No new N-glycan structures were observed in the sera of CDG-I patients as compared with normal sera. However, we observed in all subtypes a significantly increased degree of core alpha-1,6-fucosylation of the biantennary glycans as compared to normal, as well as a significant decrease in the amount of triantennary glycans. These serum N glycome changes appear to be a milder manifestation of some of the changes observed in adult liver cirrhosis patients, which is compatible with the reported steatosis and fibrosis in CDG-I patients. In the CDG-Ia subgroup, the extent of the serum N-glycome changes correlates with the aberration of the serum transferrin isoelectric focusing pattern, which measures the severity of the lack of entire N-glycan chains (primary consequence of CDG-I) in the liver and is the standard diagnostic test for this category of inherited diseases. PMID- 12626390 TI - Altered glycosylation pattern allows the distinction between prostate-specific antigen (PSA) from normal and tumor origins. AB - Prostate-specific antigen (PSA) is a glycoprotein secreted by prostate epithelial cells. PSA is currently used as a marker of prostate carcinoma because high levels of PSA are indicative of a tumor situation. However, PSA tests still suffer from a lack of specificity to distinguish between benign prostate hyperplasia and prostate cancer. To determine whether PSA glycosylation could provide a means of differentiating between PSA from normal and tumor origins, N glycan characterization of PSA from seminal fluid and prostate cancer cells (LNCaP cell line) by sequencing analysis and mass spectrometry was carried out. Glycans from normal PSA (that correspond to low and high pI PSA fractions) were sialylated biantennary complex structures, half of them being disialylated in the low pI PSA fraction and mostly monosialylated in the high pI PSA. PSA from LNCaP cells was purified to homogeneity, and its glycan analysis showed a significantly different pattern, especially in the outer ends of the biantennary complex structures. In contrast to normal PSA glycans, which were sialylated, LNCaP PSA oligosaccharides were all neutral and contained a higher fucose content. In 10 15% of the structures fucose was linked alpha1-2 to galactose, forming the H2 epitope absent in normal PSA. GalNAc was increased in LNCaP glycans to 65%, whereas in normal PSA it was only present in 25% of the structures. These carbohydrate differences allow a distinction to be made between PSA from normal and tumor origins and suggest a valuable biochemical tool for diagnosis and follow-up purposes. PMID- 12626391 TI - Molecular modeling of glycosyltransferases involved in the biosynthesis of blood group A, blood group B, Forssman, and iGb3 antigens and their interaction with substrates. AB - A terminal alpha1-3 linked Gal or GalNAc sugar residue is the common structure found in several oligosaccharide antigens, such as blood groups A and B, the xeno antigen, the Forssman antigen, and the isogloboside 3 (iGb3) glycolipid. The enzymes involved in the addition of this residue display strong amino acid sequence similarities, suggesting a common fold. From a recently solved crystal structure of the bovine alpha3-galactosyltransferase complexed with UDP, homology modeling methods were used to build the four other enzymes of this family in their locked conformation. Nucleotide-sugars, the Mn2+ ion, and oligosaccharide acceptors were docked in the models. Nine different amino acid regions are involved in the substrate binding sites. After geometry optimization of the complexes and analysis of the predicted structures, the basis of the specificities can be rationalized. In the nucleotide-sugar binding site, the specificity between Gal or GalNAc transferase activity is due to the relative size of two clue amino acids. In the acceptor site, the presence of up to three tryptophan residues define the complexity of the oligosaccharide that can be specifically recognized. The modeling study helps in rationalizing the crystallographic data obtained in this family and provides insights on the basis of substrate and donor recognition. PMID- 12626392 TI - Epitope mapping of sialyl Lewis(x) bound to E-selectin using saturation transfer difference NMR experiments. AB - A complex between sialyl Lewisx (alpha-D-Neu5Ac-[2-->3]- beta-D-Gal-[1-->4] [alpha-L-Fuc-(1-->3)]-beta-D-GlcNAc-O-[CH2]8 COOMe) and E-selectin was studied using saturation transfer difference (STD) nuclear magnetic resonance (NMR) experiments. These experiments allow the identification of the binding epitope of a ligand at atomic resolution. A semiquantitative analysis of STD total correlation spectroscopy spectra provides clear evidence that the galactose residue receives the largest saturation transfer. The protons H4 and H6 of the galactose residue are in especially close contact to the amino acids of the E selectin binding pocket. The fucose residue also receives a significant saturation transfer. The GlcNAc and Neu5Ac residues, with the exception of H3 and H3' of Neu5Ac, were found to interact weakly with the protein surface. These findings are in excellent agreement with a recently published X-ray structure and with the earlier findings from syntheses and activity assays. To further characterize the binding pocket of E-selectin, an inhibitory peptide, Ac TWDQLWDLMK-CONH2, was synthesized and the binding to E-selectin studied utilizing transfer nuclear Overhauser effect spectroscopy (trNOESY) experiments. Finally, competitive trNOESY experiments were performed, showing that the synthetic peptide is a competitive inhibitor of sialyl Lewisx. PMID- 12626393 TI - Purification and cDNA cloning of UDP-GlcNAc:GlcNAcbeta1-3Galbeta1-4Glc(NAc)-R [GlcNAc to Gal]beta1,6N-acetylglucosaminyltransferase from rat small intestine: a major carrier of dIGnT activity in rat small intestine. AB - A rat intestinal beta1,6N-acetylglucosaminyltransferase (beta1-6GnT) responsible for the formation of the beta1,6-branched poly-N-acetyllactosamine structure has been purified to apparent homogeneity by successive column chromatographic procedures using an assay wherein pyridylaminated lacto- N-triose II (GlcNAcbeta1 3Galbeta1-4Glc-PA) was used as an acceptor substrate and the reaction product was GlcNAcbeta1-3(GlcNAcbeta1-6)Galbeta1-4Glc-PA. The purified enzyme catalyzed the conversion of the polylactosamine acceptor GlcNAcbeta1-3'LacNAc into GlcNAcbeta1 3'(GlcNAcbeta1-6') LacNAc (dIGnT activity), but it could not transfer GlcNAc to LacNAcbeta1-3'LacNAc (cIGnT activity). This enzyme could also convert mucin core 1 and core 3 analogs, Galbeta1-3GalNAcalpha1-O-paranitrophenyl (pNP) and GlcNAcbeta1-3GalNAcalpha1-O-pNP, into Galbeta1-3(GlcNAcbeta1-6) GalNAcalpha1-O pNP (C2GnT activity) and GlcNAcbeta1-3(GlcNAcbeta1-6)GalNAcalpha1-O-pNP (C4GnT activity), respectively. Based on the partial amino acid sequences of the purified protein, the cDNA encoding this enzyme was cloned. The COS-1 cells transiently transfected with this cDNA had high dI/C2/C4GnT activities in a ratio of 0.34:1.00:0.90, compared with non- or mock-transfected cells. The primary structure shows a significant homology with human and viral mucin-type core 2 beta1-6GnTs (C2GnT-Ms), indicating that this enzyme is the rat ortholog of human and viral C2GnT-Ms. This is the first identification and purification of this enzyme as a major carrier of dIGnT activity in the small intestine. This rat ortholog should mostly be responsible for making distal I-branch structures on poly-N-acetyllactosamine sequences in this tissue, as well as making mucin core 2 and core 4 structures, given that it also has high C2/C4GnT activities. PMID- 12626394 TI - Characterization of carbohydrate recognition by langerin, a C-type lectin of Langerhans cells. AB - Langerin is a type II transmembrane cell surface receptor found on Langerhans cells. The extracellular domain of langerin consists of a neck region containing a series of heptad repeats and a C-terminal C-type carbohydrate-recognition domain (CRD). A role for langerin in processing of glycoprotein antigens has been proposed, but until now there has been little study of the langerin protein. In this study, analytical ultracentrifugation and circular dichroism spectroscopy of recombinant soluble fragments of human langerin have been used to show that the extracellular region of this receptor exists as a stable trimer held together by a coiled coil of alpha-helices formed by the neck region. The langerin CRD shows specificity for mannose, GlcNAc, and fucose, but only the trimeric extracellular domain fragment binds to glycoprotein ligands. Langerin extracellular domain binds mammalian high mannose oligosaccharides, as well mannose-containing structures on yeast invertase but does not bind complex glycan structures. Full length langerin stably expressed in rat fibroblast transfectants mediates efficient uptake and degradation of a mannosylated neoglycoprotein ligand. pH dependent ligand release appears to involve interactions between the CRDs or between the CRDs and the neck region in the trimer. The results are consistent with a role for langerin in internalization of both self and nonself glycoprotein antigens. PMID- 12626395 TI - The binding of human glial cell line-derived neurotrophic factor to heparin and heparan sulfate: importance of 2-O-sulfate groups and effect on its interaction with its receptor, GFRalpha1. AB - We report ELISA studies of the glycosaminoglycan binding properties of recombinant human glial cell line-derived neurotrophic factor (GDNF). We demonstrate relatively high affinity binding as soluble heparin competes with an IC50 of 0.1 micro g/ml. The binding of GDNF to heparin is particularly dependent on the presence of 2-O-sulfate groups. Highly sulfated heparan sulfate is also an effective competitor for GDNF binding. We also show that heparin at low concentrations protects GDNF from proteolytic modification by an endoprotease and also promotes the binding of GDNF to its receptor polypeptide, GFRalpha1. In both of these actions, 2-O-desulfated heparin is less effective. Considered overall, these findings provide strong support for a hypothesis that the bioactivity of GDNF during prenatal development is essentially dependent on the binding of this growth factor to 2-O-sulfate-rich heparin-related glycosaminoglycan. PMID- 12626396 TI - New insights on trehalose: a multifunctional molecule. AB - Trehalose is a nonreducing disaccharide in which the two glucose units are linked in an alpha,alpha-1,1-glycosidic linkage. This sugar is present in a wide variety of organisms, including bacteria, yeast, fungi, insects, invertebrates, and lower and higher plants, where it may serve as a source of energy and carbon. In yeast and plants, it may also serve as a signaling molecule to direct or control certain metabolic pathways or even to affect growth. In addition, it has been shown that trehalose can protect proteins and cellular membranes from inactivation or denaturation caused by a variety of stress conditions, including desiccation, dehydration, heat, cold, and oxidation. Finally, in mycobacteria and corynebacteria, trehalose is an integral component of various glycolipids that are important cell wall structures. There are now at least three different pathways described for the biosynthesis of trehalose. The best known and most widely distributed pathway involves the transfer of glucose from UDP-glucose (or GDP-glucose in some cases) to glucose 6-phosphate to form trehalose-6-phosphate and UDP. This reaction is catalyzed by the trehalose-P synthase (TPS here, or OtsA in Escherichia coli ). Organisms that use this pathway usually also have a trehalose-P phosphatase (TPP here, or OtsB in E. coli) that converts the trehalose-P to free trehalose. A second pathway that has been reported in a few unusual bacteria involves the intramolecular rearrangement of maltose (glucosyl alpha1,4-glucopyranoside) to convert the 1,4-linkage to the 1,1-bond of trehalose. This reaction is catalyzed by the enzyme called trehalose synthase and gives rise to free trehalose as the initial product. A third pathway involves several different enzymes, the first of which rearranges the glucose at the reducing end of a glycogen chain to convert the alpha1,4-linkage to an alpha,alpha1,1-bond. A second enzyme then releases the trehalose disaccharide from the reducing end of the glycogen molecule. Finally, in mushrooms there is a trehalose phosphorylase that catalyzes the phosphorolysis of trehalose to produce glucose-1-phosphate and glucose. This reaction is reversible in vitro and could theoretically give rise to trehalose from glucose-1-P and glucose. Another important enzyme in trehalose metabolism is trehalase (T), which may be involved in energy metabolism and also have a regulatory role in controlling the levels of trehalose in cells. This enzyme may be important in lowering trehalose concentrations once the stress is alleviated. Recent studies in yeast indicate that the enzymes involved in trehalose synthesis (TPS, TPP) exist together in a complex that is highly regulated at the activity level as well as at the genetic level. PMID- 12626397 TI - Alpha1,3-fucosyltransferase IX (Fut9) determines Lewis X expression in brain. AB - The expression of the Lewis X (Lex) carbohydrate structure in brain is developmentally regulated and is thought to play a role in cell-cell interaction during neuronal development. Mice possess three functional alpha1,3 fucosyltransferase genes: Fut4, Fut7, and Fut9. Fut7 is known to have no activity to synthesize Lex. In the present study, the relative activities of Fut4 and Fut9 for Lex synthesis were determined using recombinant enzymes. Fut9 exhibited very strong activity for oligosaccharide acceptors and glycolipid acceptors, that is, more than 10- and 100-fold, respectively, than that of Fut4. Furthermore, both cerebrum and cerebellum at various stages of development (E17, P0, P7, P30, P100) expressed 15-100 times more Fut9 transcript than Fut4 transcript. Neurons and astrocytes in primary culture also expressed 10-15 times more Fut9 than Fut4 transcript. Moreover, alpha1,3-Fut activity toward a polylactosamine chain in homogenates of brain tissues and primary cultured cells showed a pattern typical of Fut9, not Fut4. The developmental profile of activity for the synthesis of Lex was well correlated with that of Fut9 transcript. Immunohistochemistry with anti Fut9 monoclonal antibody revealed the distribution of the Lex structure. These results showed that Fut9 is the most responsible enzyme for the synthesis of Lex in brain. PMID- 12626398 TI - Studies on Galalpha3-binding proteins: comparison of the glycosphingolipid binding specificities of Marasmius oreades lectin and Euonymus europaeus lectin. AB - The carbohydrate binding preferences of the Galalpha3Galbeta4 GlcNAc-binding lectins from Marasmius oreades and Euonymus europaeus were examined by binding to glycosphingolipids on thin-layer chromatograms and in microtiter wells. The M. oreades lectin bound to Galalpha3-terminated glycosphingolipids with a preference for type 2 chains. The B6 type 2 glycosphingolipid (Galalpha3[Fucalpha2]Galbeta4GlcNAcbeta3Galbeta4Glcbeta1Cer) was preferred over the B5 glycosphingolipid (Galalpha3Galbeta4GlcNAcbeta3Galbeta4Glcbeta1Cer), suggesting that the alpha2-linked Fuc is accommodated in the carbohydrate binding site, providing additional interactions. The lectin from E. europaeus had broader binding specificity. The B6 type 2 glycosphingolipid was the best ligand also for this lectin, but binding to the B6 type 1 glycosphingolipid (Galalpha3[Fucalpha2]Galbeta3GlcNAcbeta3Galbeta4Glcbeta1Cer) was also obtained. Furthermore, the H5 type 2 glycosphingolipid (Fucalpha2Galbeta4GlcNAcbeta3Galbeta4Glcbeta1Cer), devoid of a terminal alpha3 linked Gal, was preferred over the the B5 glycosphingolipid, demonstrating a significant contribution to the binding affinity by the alpha2-linked Fuc. The more tolerant nature of the lectin from E. europaeus was also demonstrated by the binding of this lectin, but not the M. oreades lectin, to the x2 glycosphingolipid (GalNAcbeta3Galbeta4GlcNAcbeta3Galbeta4Glcbeta1Cer) and GlcNAcbeta3Galbeta4GlcNAcbeta3Galbeta4Glcbeta1Cer. The A6 type 2 glycosphingolipid (GalNAcalpha3[Fucalpha2]Galbeta4GlcNAcbeta3Galbeta4Glcbeta1Cer) and GalNAcalpha3Galbeta4GlcNAcbeta3Galbeta4Glcbeta1-Cer were not recognized by the lectins despite the interaction with B6 type 2 glycosphingolipid and the B5 glycosphingolipid. These observations are explained by the absolute requirement of a free hydroxyl in the 2-position of Galalpha3 and that the E. europaea lectin can accommodate a GlcNAc acetamido moiety close to this position by reorienting the terminal sugar, whereas the M. oreades lectin cannot. PMID- 12626399 TI - A transgenic insect cell line engineered to produce CMP-sialic acid and sialylated glycoproteins. AB - We have previously engineered transgenic insect cell lines to express mammalian glycosyltransferases and showed that these cells can sialylate N-glycoproteins, despite the fact that they have little intracellular sialic acid and no detectable CMP-sialic acid. In the accompanying study, we presented evidence that these cell lines can salvage sialic acids for de novo glycoprotein sialylation from extracellular sialoglycoproteins, such as fetuin, found in fetal bovine serum. This finding led us to create a new transgenic insect cell line designed to synthesize its own sialic acid and CMP-sialic acid. SfSWT-1 cells, which encode five mammalian glycosyltransferases, were transformed with two additional mammalian genes that encode sialic acid synthase and CMP-sialic acid synthetase. The resulting cell line expressed all seven mammalian genes, produced CMP-sialic acid, and sialylated a recombinant glycoprotein when cultured in a serum-free growth medium supplemented with N-acetylmannosamine. Thus the addition of mammalian genes encoding two enzymes involved in CMP-sialic acid biosynthesis yielded a new transgenic insect cell line, SfSWT-3, that can sialylate recombinant glycoproteins in the absence of fetal bovine serum. This new cell line will be widely useful as an improved host for baculovirus-mediated recombinant glycoprotein production. PMID- 12626400 TI - The dendritic cell-specific C-type lectin DC-SIGN is a receptor for Schistosoma mansoni egg antigens and recognizes the glycan antigen Lewis x. AB - Schistosoma mansoni soluble egg antigens (SEAs) are crucially involved in modulating the host immune response to infection by S. mansoni. We report that human dendritic cells bind SEAs through the C-type lectin dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN). Monoclonal antibodies against the carbohydrate antigens Lewisx (Lex) and GalNAcbeta1-4(Fucalpha1-3)GlcNAc (LDNF) inhibit binding of DC-SIGN to SEAs, suggesting that these glycan antigens may be critically involved in binding. In a solid-phase adhesion assay, DC-SIGN-Fc binds polyvalent neoglycoconjugates that contain the Lex antigen, whereas no binding was observed to Galbeta1-4GlcNAc, and binding to neoglycoconjugates containing only alpha-fucose or oligosaccharides with a terminal alpha1-2-linked fucose is low. These data indicate that binding of DC-SIGN to Lex antigen is fucose dependent and that adjacent monosaccharides and/or the anomeric linkage of the fucose are important for binding activity. Previous studies have shown that DC SIGN binds HIV gp120 that contains high-mannose-type N-glycans. Site-directed mutagenesis within the carbohydrate recognition domain (CRD) of DC-SIGN demonstrates that amino acids E324 and E347 are involved in binding to HIV gp120, Lex, and SEAs. By contrast, mutation of amino acid Val351 abrogates binding to SEAs and Lex but not HIV gp120. These data suggest that DC-SIGN recognizes these ligands through different (but overlapping) regions within its CRD. Our data imply that DC-SIGN not only is a pathogen receptor for HIV gp120 but may also function in pathogen recognition by interaction with the carbohydrate antigens Lex and possibly LDNF, which are found on important human pathogens, such as schistosomes and the bacterium Helicobacter pylori. PMID- 12626401 TI - Evidence for a sialic acid salvaging pathway in lepidopteran insect cells. AB - We previously described a transgenic insect cell line, Sfbeta4GalT/ST6, that expresses mammalian beta-1,4-galactosyltransferase and alpha2,6-sialyltransferase genes and produces glycoproteins with terminally sialylated N-glycans. The ability of these cells to produce sialylated N-glycans was surprising because insect cells contain only small amounts of sialic acid and no detectable CMP sialic acid. Thus, it was of interest to investigate potential sources of sialic acids for sialoglycoprotein synthesis by these cells. We found that Sfbeta4GalT/ST6 cells can produce sialylated N-glycans when cultured in the presence but not in the absence of fetal bovine serum. The serum component(s) supporting N-glycan sialylation by Sfbeta4GalT/ST6 cells is relatively large-it was not removed by dialysis in a 50,000-molecular-weight cutoff membrane. Serum free media supplemented with purified fetuin but not asialofetuin supported N glycan sialylation by Sfbeta4GalT/ST6 cells. The terminally sialylated N-glycans isolated from fetuin also supported glycoprotein sialylation by Sfbeta4GalT/ST6 cells. Finally, serum-free medium supplemented with N-acetylneuraminic acid or N acetylmannosamine supported glycoprotein sialylation by Sfbeta4GalT/ST6 cells but to a much lower degree than serum or fetuin. These results provide the first evidence of a sialic acid salvaging pathway in insect cells, which begins to explain how Sfbeta4GalT/ST6 and other transgenic insect cell lines can sialylate recombinant glycoproteins in the absence of a more obvious source of CMP-sialic acid. PMID- 12626402 TI - Sulfated fucans, fresh perspectives: structures, functions, and biological properties of sulfated fucans and an overview of enzymes active toward this class of polysaccharide. AB - Sulfated fucans, frequently referred to simply as fucans, constitute a class of polysaccharides first isolated in 1913. For many years fucans were regarded only as a potential source of l-fucose, although their anticoagulant activity was known. Even as the potent effects of fucans on physiological systems have become better characterized, structural studies have lagged behind. Recently the search for new drugs has raised increased interest in sulfated fucans. In the past few years, several structures of algal and invertebrate fucans have been solved, and many aspects of their biological activity have been elucidated. From this work emerges a more interesting picture of this class of polysaccharides than was previously suspected. The availability of purified fucans and fucan fractions with simple, but varied structures, in conjunction with the development of new enzymatic tools, demonstrate that the biological properties of sulfated fucans are not only a simple function of their charge density but also are determined by detailed structural features. PMID- 12626403 TI - Characterization of the rat alpha(1,3)galactosyltransferase: evidence for two independent genes encoding glycosyltransferases that synthesize Galalpha(1,3)Gal by two separate glycosylation pathways. AB - The important xenoepitope Galalpha(1,3)Gal was thought to be exclusively synthesized by a single alpha(1,3)galactosyltransferase. However, the cloning of the distant family member rat iGb3 synthase, which is also capable of synthesizing Galalpha(1,3)Gal as the glycolipid structure iGb3, challenges the notion that alpha(1,3)galactosyltransferase is the sole Galalpha(1,3)Gal synthesizing enzyme. We describe the cloning of the rat homolog of alpha(1,3)galactosyltransferase, showing that indeed the rat expresses two distinct alpha(1,3)galactosyltransferases, alpha(1,3)GT and iGb3 synthase. Rat alpha(1,3)galactosyltransferase shows a high amino acid sequence identity with the alpha(1,3)galactosyltransferase of mouse (90%), pig (76%), and ox (75%), in contrast to the low amino acid sequence identity (42%) with iGb3 synthase. The rat alpha(1,3)galactosyltransferase is expressed in heart, brain, spleen, kidney, and liver and has a similar intron/exon structure to the mouse alpha(1,3)galactosyltransferase. Transfection studies show that in contrast to the iGb3 synthase, rat alpha(1,3)galactosyltransferase can synthesize Galalpha(1,3)Gal on glycoproteins but cannot synthesize the glycolipid iGb3, defining two separate glycosylation pathways for the synthesis of Galalpha(1,3)Gal. Furthermore iGb3 synthase was found to be distinct from alpha(1,3)GT with its ability to synthesize poly-alpha-Gal glycolipid structures. PMID- 12626404 TI - Structures of the glycosylphosphatidylinositol membrane anchors from Aspergillus fumigatus membrane proteins. AB - Glycosylphosphatidylinositol (GPI)-anchored proteins have been identified in all eukaryotes. In fungi, structural and biosynthetic studies of GPIs have been restricted to the yeast Saccharomyces cerevisiae. In this article, four GPI anchored proteins were purified from a membrane preparation of the human filamentous fungal pathogen Aspergillus fumigatus. Using new methodology applied to western blot protein bands, the GPI structures were characterized by ES-MS, fluorescence labeling, HPLC, and specific enzymatic digestions. The phosphatidylinositol moiety of the A. fumigatus GPI membrane anchors was shown to be an inositol-phosphoceramide containing mainly phytosphingosine and monohydroxylated C24:0 fatty acid. In constrast to yeast, only ceramide was found in the GPI anchor structures of A. fumigatus, even for Gel1p, a homolog of Gas1p in S. cerevisiae that contains diacylglycerol. The A. fumigatus GPI glycan moiety is mainly a linear pentomannose structure linked to a glucosamine residue: Manalpha1-3Manalpha1-2Manalpha1-2Manalpha1-6Manalpha1-4GlcN. PMID- 12626405 TI - A novel GlcNAcalpha1-HPO3-6Gal(1-1)ceramide antigen and alkylated inositol phosphoglycerolipids expressed by the liver fluke Fasciola hepatica. AB - The acidic (glyco)lipids of the parasitic liver fluke Fasciola hepatica exhibited two different phosphate-containing species, designated AL-I and AL-II, which were analyzed by MALDI-TOF MS, ESI MS, NMR, methylation analysis, and combined GC-MS in conjunction with HF treatment. AL-I was structurally determined as 1-O hexadecyl-sn-glycerol-3-phosphoinositol, an ether bond variant of lysophosphatidylinositol. The structure of AL-II was shown to be GlcNAcalpha1 HPO3-6Gal(1-1)ceramide. Ceramide analysis revealed as major components 2 hydroxyoctadecanoic acid [18:0(2-OH)] together with C18- and C20 phytosphingosines. AL-II was apparently highly antigenic and strongly recognized by both animal- and human-F. hepatica infection sera. Furthermore, inhibition ELISAs revealed that the unusual antigenic determinant GlcNAcalpha1-HPO3- phosphate might have a potential in the serodiagnosis of F. hepatica infections. PMID- 12626406 TI - Heparin sequencing. AB - Heparin is a highly sulfated glycosaminoglycan widely used as an anticoagulant. Modifications in its relatively uniform structure appear to be key to its recognition and modulation of serine proteases, growth factors, chemokines, and extracellular proteins, as has been most clearly demonstrated in the antithrombin binding site. We sequenced the major oligosaccharides released from mastocytoma heparin by partial nitrous acid using a highly sensitive technique tailored for sequencing of metabolically radiolabeled heparin. It utilizes partial nitrous acid cleavage to allow simultaneous sequencing of the internal components of the oligosaccharide under investigation by specific lysosomal exoenzymes. Sequencing revealed that although the majority of the heparin disaccharides are N-, 2-O-, and 6-O-sulfated, the less sulfated disaccharides (lacking 2-O- or 6-O-sulfates) seem to be spaced out along the chain. The technique may be particularly useful for characterizing heparin from novel sources, such as the glial progenitor cells and Ascidia, as well as for sequencing protein binding sites. PMID- 12626407 TI - Characterization by NMR and molecular modeling of the binding of polyisoprenols and polyisoprenyl recognition sequence peptides: 3D structure of the complexes reveals sites of specific interactions. AB - The objective of these studies was to test the hypothesis that proteins that contain potential polyisoprenyl recognition sequences (PIRSs) in their transmembrane-spanning domain can bind to the polyisoprenyl (PI) glycosyl carrier lipids undecaprenyl phosphate (C55-P) and dolichyl phosphate (C95-P). A number of prokaryotic and eukaryotic glycosyltransferases that utilize PI coenzymes contain a conserved PIRS postulated to be the active PI binding domain. To study this problem, we first determined the 3D structure of a PIRS peptide, NeuE, by homonuclear 2D 1H-nuclear magnetic resonance (NMR) spectroscopy. Experimentally generated distance constraints derived from nuclear Overhauser enhancement and torsion angle constraints derived from coupling constants were used for restrained molecular dynamics and energy minimization calculations. Molecular models of the NeuE peptide were built based on calculations of energy minimization using the DGII program NMRchitect. 3D models of dolichol (C95) and C95-P were built based on our 2D 1H-NMR nuclear Overhauser enhancement spectroscopy (NOESY) results and refined by energy minimization with respect to all atoms using the AMBER (assisted modeling with energy refinements) force field. Our energy minimization studies were carried out on a conformational model of dolichol that was originally derived from small-angle X-ray scattering and molecular mechanics methods. These results revealed that the PIs are conformationally nearly identical tripartite molecules, with their three domains arranged in a coiled, helical structure. Analyses of the intermolecular cross peaks in the 2D NOESY spectra of PIRS peptides in the presence of PIs confirmed a highly specific interaction and identified key contact amino acids in the NeuE peptide that constituted a binding motif for interacting with the PIs. These studies also showed that subtle conformational changes occurred within both the PIs and the NeuE peptide after binding. 3D structures of the resulting molecular complexes revealed that each PI could bind more than one PIRS peptide. These studies thus represent the first evidence for a direct physical interaction between specific contact amino acids in the PIRS peptides and the PIs and supports the hypothesis of a bifunctional role for the PIs. The central idea is that these superlipids may serve as a structural scaffold to organize and stabilize in functional domains PIRS-containing proteins within multiglycosyltransferase complexes that participate in biosynthetic and translocation processes. PMID- 12626409 TI - (Arg)3 within the N-terminal domain of glucosidase I contains ER targeting information but is not required absolutely for ER localization. AB - Glucosidase I is an endoplasmic reticulum (ER) type II membrane enzyme that cleaves the distal alpha1,2-glucose of the asparagine-linked GlcNAc2-Man9-Glc3 precursor. To identify sequence motifs responsible for ER localization, we prepared a protein chimera by transferring the cytosolic and transmembrane domain of glucosidase I to the luminal domain of Golgi-Man9-mannosidase. The GIM9 hybrid was overexpressed in COS 1 cells as an ER-resident protein that displayed alpha1,2-mannosidase activity, excluding the possibility that the glucosidase I specific domains interfere with folding of the Man9-mannosidase catalytic domain. After substitution of the Args in position 7, 8, or 9 relative to the N-terminus by leucine, the GIM9 mutants were transported to the cell surface indicating that the (Arg)3 sequence functions as an ER-targeting motif. Cell surface expression was also observed after substitution of Arg-7 or Arg-8 but not Arg-9 in GIM9 by either lysine or histidine. Thus the side chain structure, including its positive charge, appears to be essential for signal function. Analysis of the N-linked glycans suggests that the (Arg)3 sequence mediates ER localization through Golgi to-ER retrograde transport. Glucosidase I remained localized in the ER after truncation or mutation of the N-terminal (Arg)3 signal, in contrast to comparable GIM9 mutants. ER localization was also observed with an M9GI chimera consisting of the cytosolic and transmembrane domain of Man9-mannosidase and the glucosidase I catalytic domain. ER-specific targeting information must therefore be provided by sequence motifs contained within the glucosidase I luminal domain. This structural information appears to direct ER localization by retention rather than by retrieval, as concluded from N-linked Man9-GlcNAc2 being the major glycan released from the wild-type enzyme. PMID- 12626408 TI - Opposite effects of galectin-1 on alternative metabolic pathways of L-arginine in resident, inflammatory, and activated macrophages. AB - Recent evidence has implicated galectins and their carbohydrate ligands as master regulators of the inflammatory response. Galectin-1, a member of this family, has shown specific anti-inflammatory and immunoregulatory effects. To gain insight into the potential mechanisms involved in these effects, we investigated the effects of galectin-1 in L-arginine metabolism of peritoneal rat macrophages. Pretreatment of macrophages with galectin-1 resulted in a dose- and time dependent inhibition of lipopolysaccharide-induced nitric oxide (NO) production, accompanied by a decrease in inducible nitric oxide synthase (iNOS) expression (the classic pathway of L-arginine). On the other hand, galectin-1 favored the balance toward activation of L-arginase, the alternative metabolic pathway of L arginine. Inhibition of NO production was not the result of increased macrophage apoptosis because addition of this beta-galactoside-binding protein to macrophages under the same experimental conditions did not affect the apoptotic threshold of these cells. To understand how endogenous galectin-1 is regulated in macrophages under inflammatory stress, we finally explored the ultrastructural distribution, expression, and secretion of galectin-1 in resident, inflammatory, and activated macrophages. This study provides an alternative cellular mechanism based on the modulation of L-arginine metabolism to understand the molecular basis of the anti-inflammatory properties displayed by this carbohydrate-binding protein. PMID- 12626410 TI - In vitro oligosaccharide synthesis using intact yeast cells that display glycosyltransferases at the cell surface through cell wall-anchored protein Pir. AB - A glycosyltransferase was fused to the yeast cell wall protein Pir, which forms the Pir1-4 protein family and is incorporated into the cell wall by an unknown linkage to be displayed at the yeast cell surface. We first expressed the PIR1-HA gma12+ fusion, in which gma12+ encodes alpha-1,2-galactosyltransferase from the fission yeast Schizosaccharomyces pombe under the Saccharomyces cerevisiae GAPDH promoter. The alpha-1,2-galactosyltransferase activity was detected at the surface of the intact cells that produce Pir1-HA-Gma12 fusion. To further demonstrate sequential oligosaccharide synthesis, two plasmids containing PIR1-HA KRE2 and PIR2-FLAG-MNN1 fusion genes were constructed in which KRE2 and MNN1 encode alpha-1,2-mannosyltransferase and alpha-1,3-mannosyltransferase from S. cerevisiae, respectively. The intact yeast cells transformed with these two plasmids added mannoses initially with an alpha-1,2 linkage and subsequently with an alpha-1,3 linkage to the alpha-1,2-mannobiose acceptor in the presence of a GDP-mannose donor, demonstrating that Pir1 and Pir2 can be used as anchors to simultaneously immobilize several glycosyltransferases at the yeast cell surface. Based on the high acceptor specificity of glycosyltransferases, we propose a simple in vitro method for oligosaccharide synthesis using the yeast intact cell as a biocatalyst. PMID- 12626411 TI - The two rat alpha 2,6-sialyltransferase (ST6Gal I) isoforms: evaluation of catalytic activity and intra-Golgi localization. AB - alpha2,6-Sialyltransferase (ST6Gal I) functions in the Golgi to terminally sialylate the N-linked oligosaccharides of glycoproteins. Interestingly, rat ST6Gal I is expressed as two isoforms, STtyr and STcys, that differ by a single amino acid in their catalytic domains. In this article, our goal was to evaluate more carefully possible differences in the catalytic activity and intra-Golgi localization of the two isoforms that had been suggested by earlier work. Using soluble recombinant STtyr and STcys enzymes and three asialoglycoprotein substrates for in vitro analysis, we found that the STcys isoform was somewhat more active than the STtyr isoform. However, we found no differences in isoform substrate choice when these proteins were expressed in Chinese hamster ovary cells, and sialylated substrates were detected by lectin blotting. Immuno fluorescence and immunoelectron microscopy revealed differences in the relative levels of the isoforms found in the endoplasmic reticulum (ER) and Golgi of transiently expressing cells but similar intra-Golgi localization. STtyr was restricted to the Golgi in most cells, and STcys was found in both the ER and Golgi. The ER localization of STcys was especially pronounced with a C-terminal V5 epitope tag. Ultrastructural and deconvolution studies of immunostained HeLa cells expressing STtyr or STcys showed that within the Golgi both isoforms are found in medial-trans regions. The similar catalytic activities and intra-Golgi localization of the two ST6Gal I isoforms suggest that the particular isoform expressed in specific cells and tissues is not likely to have significant functional consequences. PMID- 12626412 TI - The effects of ethanol on the glycosylation of human transferrin. AB - Appearance of a hyposialylated transferrin fraction in the plasma during chronic alcohol exposure is a well-known phenomenon, and it represents the best available marker of chronic alcohol consumption. The mechanisms of its appearance are still not well understood and are extremely complex, involving biosynthesis and catabolism alterations, although the only structural abnormality described corresponds to the loss of an entire glycan chain. We analyzed and compared the oligosaccharides present on the different isoforms of purified transferrin isolated from control and patients with severe alcohol abuse by fluorescent carbohydrate electrophoresis and matrix-assisted laser desorption ionization mass spectrometry. Our data indicate that the major modification observed is the loss of an entire oligosaccharide chain; we also demonstrate that there is a modification of terminal sialylation. Carbohydrate-deficient transferrin (CDT) is the result of multiple alterations of glycosylation. These results give a partial explanation to the poor sensitivity of the measurement of CDT and its controversial use as a marker of chronic alcohol consumption. PMID- 12626413 TI - Evolution of substrate recognition across a multigene family of glycosyltransferases in Arabidopsis. AB - The complete sequence of the Arabidopsis genome enables definitive characterization of multigene families and analysis of their phylogenetic relationships. Using a consensus sequence previously defined for glycosyltransferases that use small-molecular-weight acceptors, 107 gene sequences were identified in the Arabidopsis genome and used to construct a phylogenetic tree. Screening recombinant proteins for their catalytic activities in vitro has revealed enzymes active toward physiologically important substrates, including hormones and secondary metabolites. The aim of this study has been to use the phylogenetic relationships across the entire family to explore the evolution of substrate recognition and regioselectivity of glucosylation. Hydroxycoumarins have been used as the model substrates for the analysis in which 90 sequences have been assayed and 48 sequences shown to recognize these compounds. The study has revealed activity in 6 of the 14 phylogenetic groups of the multigene family, suggesting that basic features of substrate recognition are retained across substantial evolutionary periods. PMID- 12626414 TI - Activities and expression pattern of the carbohydrate sulfotransferase GlcNAc6ST 3 (I-GlcNAc6ST): functional implications. AB - In recent years, a family of five GlcNAc-6-O-sulfotransferases, called the GlcNAc6STs, has been molecularly cloned. One of these, GlcNAc6ST-2 (originally named HEC-GlcNAc6ST or LSST), shows a very restricted expression at the mRNA level in high endothelial cells (HECs) of lymph nodes high endothelial venules (HEVs). This enzyme has been shown to be involved in elaborating the 6-sulfo sLex structure on a set of mucin-like acceptors within HECs, thus providing a critical recognition determinant for L-selectin during the process of lymphocyte homing to lymph nodes. Limited information has been available about the closely related sulfotransferase known as GlcNAc6ST-3 (I-GlcNAc6ST). Here, employing transfection experiments with a series of glycoprotein acceptors, we report that this sulfotransferase has a marked preference for sulfating O-linked sugars of mucin type acceptors, whereas other sulfotransferases in the family (GlcNAc6ST-1, GlcNAc6ST-2) and a Gal-6-O-sulfotransferase exhibit strong activity on both mucin type acceptors and glycoproteins with predominantly N-linked chains. PCR analysis of cDNAs derived from a panel of tissues and purified cell populations confirms the strong expression of GlcNAc6ST-3 in gut-associated tissues and extends the expression to include lymphocytes. In contrast to GlcNAc6ST-2, GlcNAc6ST-3 transcripts are present minimally, if at all, in HECs; moreover, this enzyme is not able to generate the 6-sulfo sLex epitope in transfected cells. These latter findings argue that GlcNAc6ST-3 is not involved in generating HEV-expressed ligands for L-selectin. PMID- 12626415 TI - Glycosylation of human pancreatic ribonuclease: differences between normal and tumor states. AB - Characterization of the N-glycans from human pancreatic ribonuclease (RNase 1) isolated from healthy pancreas and from pancreatic adenocarcinoma tumor cells (Capan-1 and MDAPanc-3) revealed completely different glycosylation patterns. RNase 1 from healthy cells contained neutral complex biantennary structures, with smaller amounts of tri- and tetraantennary compounds, and glycans with poly-N acetyllactosamine extensions, all extensively fucosylated. In contrast, RNase 1 glycans from tumor cells (Capan-1) were fucosylated hybrid and complex biantennary glycans with GalNAc-GlcNAc antennae. RNase 1 glycans from Capan-1 and MDAPanc-3 cells also contained sialylated structures completely absent in the healthy pancreas. Some of these features provide distinct epitopes that were clearly detected using monoclonal antibodies against carbohydrate antigens. Thus monoclonal antibodies to Lewis(y) reacted only with normal pancreatic RNase 1, whereas, in contrast, monoclonal antibodies to sialyl-Lewis(x) and sialyl Lewis(a) reacted only with RNase 1 secreted from the tumor cells. These glycosylation changes in a tumor-secreted protein, which reflect fundamental changes in the enzymes involved in the glycosylation pathway, open up the possibility of using serum RNase 1 as a tumor marker of pancreatic adenocarcinoma. PMID- 12626416 TI - Glycosylation of human recombinant gonadotrophins: characterization and batch-to batch consistency. AB - The glycan moiety of human recombinant gonadotrophins (r-hFSH, r-hLH, and r-hCG) produced in CHO cell lines has been characterized by a combination of chromatographic and mass spectrometric techniques, including both matrix-assisted laser desorption ionization and electrospray. Two glycan mapping methods have been developed for the three gonadotrophins that allow separation of the glycans according to either their charge or sialylation level or their antennarity. A method was also developed for r-hCG that permits the complete resolution of the N glycan from the O-glycan species. Whereas the structure found for the N-glycans of the gonadotrophins was in agreement with the complex type model, the structure for an O-glycan of r-hCG, not yet described, has been unambiguously determined using nanoelectrospray ion trap mass spectrometry. Using these two glycan mapping methods, the high level of batch-to-batch consistency achieved for the glycosylation of the three recombinant gonadotrophins in commercial production has been shown. These data demonstrate the tight control that can be achieved in the manufacturing of complex recombinant therapeutic glycoproteins, which is a prerequisite to the delivering of a guaranteed dose of drug from vial to vial, and in turn to ensuring the clinical efficacy of the product. PMID- 12626417 TI - Plant cultured cells expressing human beta1,4-galactosyltransferase secrete glycoproteins with galactose-extended N-linked glycans. AB - Previously, we generated transgenic tobacco BY2 suspension-cultured cells (GT6 cells) that produced human beta1,4-galactosyltransferase. In this study, we analyze the N-glycan structures of glycoproteins secreted from GT6 cells to the spent medium. The N-glycans were liberated by hydrazinolysis, and the resulting oligosaccharides were labeled with 2-aminopyridine (PA). The pyridylaminated glycans were purified by reversed-phase and size-fractionation HPLC. The structures of the PA sugar chains were identified by the combined use of 2D PA sugar chain mapping, MS/MS analysis, and exoglycosidase digestion. The distribution of proposed N-glycan structures of GT6-secreted glycoproteins (GalGNM5 [26.8%], GalGNM4 [18.4%], GalGNM3 [19.6%], and GalGNM3X [35.2%]) is different from that found in intracellular glycoproteins (M7A [9.3%], M7B [15.9%], M6B [19.5%], M5 [1.4%], M3X [6.6%], GalGNM5 [35.5%], and GalGNM3 [11.8%]). In vitro, sialic acid was transferred to sugar chains of extracellular glycoproteins from the GT6 spent medium. The results suggest that sugar chains of extracellular glycoproteins from the GT6 spent medium are candidates for substrates of sialic acid transfer. PMID- 12626418 TI - Sialylation and sulfation of lactosylceramide distinctly regulate anchorage independent growth, apoptosis, and gene expression in 3LL Lewis lung carcinoma cells. AB - To investigate the significance of sialylation and sulfation of lactosylceramide in transformed cells, we established ganglioside GM3- and lactosylsulfatide (SM3) reconstituted cells by transfecting cDNAs of GM3 synthase and cerebroside sulfotransferase into the J5 subclone of 3LL Lewis lung carcinoma cells. The J5 clone was selected for the transfection of these genes because it lacks GM3 and SM3 but accumulates lactosylceramide. The anchorage-dependent growth of both GM3- and SM3-reconstituted cells was similar. However, anchorage-independent growth (as measured by colony-forming ability in soft agar) of the SM3- reconstituted cells was almost completely lost, which supports our previous observation showing the suppression of tumorigenic potential in vivo and beta1 integrin gene expression induced by the introduction of cerebroside sulfotransferase gene (Kabayama et al. [2001] J. Biol. Chem., 276, 26777-26783). The GM3-reconstituted cells formed a significantly higher number of colonies in soft agar compared to mock-transfected cells and began to proliferate and become resistant to apoptosis when serum was depleted, indicating that endogenous GM3 is essential for maintaining these fundamental properties of malignant cells. We also found that serum-induced ERK1/2 activation was suppressed in the GM3-reconstituted cells, suggesting that anchorage-independent cell cycle initiation by endogenous GM3 is elicited through pathway(s) independent of ERK1/2 activation. The selective down regulation of platelet-derived growth factor (PDGF)-dependent ERK1/2 activation in the GM3-reconstituted cells was due to the substantial decreases of PDGF alpha receptor mRNA and protein, but in the SM3-reconstituted cells PDGF alpha receptor expression was similar to mock cells. Thus, endogenously produced GM3 and SM3 differentially and distinctly regulate tumor-progression ability, that is, GM3 leads the transformed phenotype of J5 cells to promotion and SM3 to abrogation. PMID- 12626419 TI - Structural determination of the N-glycans of a lepidopteran arylphorin reveals the presence of a monoglucosylated oligosaccharide in the storage protein. AB - The structures of the oligosaccharides attached to arylphorin from Chinese oak silkworm, Antheraea pernyi, have been determined. Arylphorin, a storage protein present in fifth larval hemolymph, contained 4.8% (w/w) of carbohydrate that was composed of Fuc:GlcNAc:Glc:Man=0.2:4.0:1.4:13.6 moles per mole protein. Four moles of GlcNAc in oligomannose-type oligosaccharides strongly suggest that the protein contains two N-glycosylation sites. Normal-phase HPLC and mass spectrometry oligosaccharide profiles confirmed that arylphorin contained mainly oligomannose-type glycans as well as truncated mannose-type structures with or without fucosylation. Interestingly, the most abundant oligosaccharide was monoglucosylated Man9-GlcNAc2, which was characterized by normal-phase HPLC, mass spectrometry, Aspergillus saitoi alpha-mannosidase digestion, and 1H 600 MHz NMR spectrometry. This glycan structure is not normally present in secreted mammalian glycoproteins; however, it has been identified in avian species. The Glc1Man9GlcNAc2 structure was present only in arylphorin, whereas other hemolymph proteins contained only oligomannose and truncated oligosaccharides. The oligosaccharide was also detected in the arylphorin of another silkworm, Bombyx mori, suggesting a specific function for the Glc1Man9GlcNAc2 glycan. There were no processed glucosylated oligosaccharides such as Glc1Man5-8GlcNAc2. Furthermore, Glc1Man9GlcNAc2 was not released from arylophorin by PNGase F under nondenaturing conditions, suggesting that the N-glycosidic linkage to Asn is protected by the protein. Glc1Man9GlcNAc2 may play a role in the folding of arylphorin or in the assembly of hexamers. PMID- 12626420 TI - Immunoreactivity in mammals of two typical plant glyco-epitopes, core alpha(1,3) fucose and core xylose. AB - The presence of nonmammalian core alpha(1,3)-fucose and core xylose glyco epitopes on glycans N-linked to therapeutic glycoproteins produced in plants has raised the question of their immunogenicity in human therapy. We address this question by studying the distribution of these N-glycans in pea, rice, and maize (which are the crops intended for the production of therapeutic proteins) and by reinvestigating their immunogenicity in rodents. We found that immunization with a model glycoprotein, horseradish peroxidase, elicits in C57BL/6 mice and rats the production of antibodies (Abs) specific for core alpha(1,3)-fucose and core xylose epitopes. Furthermore, we demonstrated that about 50% of nonallergic blood donors contains in their sera Abs specific for core xylose, whereas 25% have Abs against core alpha(1,3)-fucose. These Abs probably result from sensitization to environmental antigens. Although the immunological significance of these data is too speculative at the moment, the presence of such Abs might introduce some limitations to the use of plant-derived biopharmaceutical glycoproteins, such as an accelerated clearance during human therapy. PMID- 12626421 TI - Schistosoma mansoni-infected mice produce antibodies that cross-react with plant, insect, and mammalian glycoproteins and recognize the truncated biantennaryN glycan Man3GlcNAc2-R. AB - To reveal the role of cross-reactive carbohydrate determinants in the host immune response in helminth infections and allergenicity, we developed monoclonal antibodies (mAbs) that recognize glycan epitopes present on glycoconjugates from both helminths and plants. An IgM mAb (100-4G11-A) was selected from a panel of anti-glycan mAbs generated from Schistosoma-infected or immunized mice because it recognized both a plant glycoprotein horseradish peroxidase and phospholipase A2 from honeybee venom. On further characterization, it was shown that mAb 100-4G11 A recognizes the truncated biantennary N-glycan Man3GlcNAc2-R. Immunocytochemical analysis and immunoblotting with this mAb demonstrated that Man3GlcNAc2-R structures occur on many glycoproteins of schistosomes and other invertebrates. Remarkably, Man3GlcNAc2-R is also expressed on a restricted number of vertebrate glycoproteins. Our data indicate that this truncated N-glycan is immunogenic in mice during the course of infection. Nevertheless, no elevated antibody levels against this glycan epitope could be detected in sera of individuals infected with Schistosoma mansoni. PMID- 12626422 TI - The underglycosylation of plasma alpha 1-antitrypsin in congenital disorders of glycosylation type I is not random. AB - Conditions under which the glycosylation capacity of cells is limited provide an opportunity for studying the efficiency of site-specific glycosylation and the role of glycosylation in the maturation of glycoproteins. Congenital disorders of glycosylation type 1 (CDG-I) provide such a system. CDG-I is characterized by underglycosylation of glycoproteins due to defects in the assembly or transfer of the common dolichol-pyrophosphate-linked oligosaccharide precursor of asparagine linked glycans. Human plasma alpha1-antitrypsin is normally fully glycosylated at three asparagine residues (46, 83, and 247), but un-, mono-, di-, and fully glycosylated forms of alpha1-antitrypsin were detected by 2D PAGE in the plasma from patients with CDG-I. The state of glycosylation of the three asparagine residues was analyzed in all the underglycosylated forms of alpha1-antitrypsin by peptide mass fingerprinting using matrix-assisted laser desorption ionization time-of-flight mass spectrometry. It was found that asparagine 46 was always glycosylated and that asparagine 83 was never glycosylated in the underglycosylated glycoforms of alpha1-antitrypsin. This showed that the asparagine residues are preferentially glycosylated in the order 46>247>83 in the mature underglycosylated forms of alpha1-antitrypsin found in plasma. It is concluded that the nonoccupancy of glycosylation sites is not random under conditions of decreased glycosylation capacity and that the efficiency of glycosylation site occupancy depends on structural features at each site. The implications of this observation for the intracellular transport and sorting of glycoproteins are discussed. PMID- 12626423 TI - Identification and characterization of adsorbed serum sialoglycans on Leishmania donovani promastigotes. AB - Sialic acids as terminal residues of oligosaccharide chains play a crucial role in several cellular recognition events. The presence of sialic acid on promastigotes of Leishmania donovani, the causative organism of Indian visceral leishmaniasis, was demonstrated by fluorimetric high-performance liquid chromatography showing Neu5Ac and, to a minor extent, Neu5,9Ac2. The presence of Neu5Ac was confirmed by GC/MS analysis. Furthermore, binding with sialic acid binding lectins Sambucus nigra agglutinin (SNA), Maackia amurensis agglutinin (MAA), and Siglecs showed the presence of both alpha2,3- and alpha2,6-linked sialic acids. No endogenous biosynthetic machinery for Neu5Ac could be demonstrated in the parasite. Concomitant western blotting of parasite membranes and culture medium with SNA demonstrated the presence of common sialoglyconjugates (123, 90, and 70 kDa). Similarly, binding of MAA with parasite membrane and culture medium showed three analogous sialoglycans corresponding to 130, 117, and 70 kDa, indicating that alpha2,3- and alpha2,6-linked sialoglycans are adsorbed from the fetal calf serum present in the culture medium. L. donovani promastigotes also reacted with Achatinin-H, a lectin that preferentially identifies 9-O-acetylated sialic acid in alpha2-->6 GalNAc linkage. This determinant was evidenced on parasite cell surfaces by cell agglutination, ELISA, and flow cytometry, where its binding was abolished by pretreatment of cells with a recombinant 9-O-acetylesterase derived from the HE1 region of the influenza C esterase gene. Additionally, binding of CD60b, a 9-O-acetyl GD3-specific monoclonal antibody, corroborated the presence of terminal 9-O-acetylated disialoglycans. Our results indicate that sialic acids (alpha2-->6 and alpha2-->3 linked) and 9-O-acetyl derivatives constitute components of the parasite cell surface. PMID- 12626424 TI - The contribution of tandem repeat number to the O-glycosylation of mucins. AB - The serine- and threonine-rich tandem repeat (TR) units that make up the characteristic feature of mucin glycoproteins are often polymorphic with substantial genetic variation in TR number. The precise effect of TR number on O glycosylation is not fully understood, although the TR number of several mucins may be associated with apparent susceptibility to certain human diseases. To evaluate the contribution of TR number to O-glycosylation, we generated a series of chimeric mucins carrying increasing numbers of TR units from the MUC5B mucin in the context of an epitope-tagged MUC1 mucin backbone. These mucins were expressed in Caco2 colon carcinoma cell clones and purified by immunoprecipitation. O-Glycosylation was investigated by western blotting with antibodies to known carbohydrate structures and by fast atom bombardment-mass spectrometry. Additional carbohydrate epitopes were detected with antibodies on chimeric mucins with a higher TR number in comparison to those with fewer TRs. Using mass spectrometry, higher-molecular-weight glycans were detected more frequently on the mucins with extended TRs compared to those with fewer TRs. However no novel carbohydrate structures were seen, suggesting that TR number does not affect the specificity of O-glycosylation. PMID- 12626425 TI - Purification and molecular identification of the human hyaluronan receptor for endocytosis. AB - The clearance of hyaluronan (HA) and chondroitin sulfates from the circulating blood and lymph in the body is mediated by the membrane-bound HA receptor for endocytosis (HARE). Previously, we found that two HARE species of approximately 175 kDa and approximately 300 kDa are abundant in the sinusoidal endothelial cells in rat liver, spleen, and lymph nodes (Zhou et al. [2000], J. Biol. Chem., 275, 37733-37741). In the present study, immunocytochemical analysis of human tissues showed a similar pattern with abundant expression of HARE in the sinusoidal endothelial cells of human liver, spleen, and lymph nodes. The two human HARE proteins were immunoaffinity-purified from human spleen. Each protein was recognized in western blots using several anti-rat HARE monoclonal antibodies and was able to bind 125I-HA specifically. In nonreducing SDS-PAGE, these two human HARE species migrated at approximately 190 kDa and approximately 315 kDa; both proteins are approximately 15 kDa larger than the corresponding rat HAREs, although the de-N-glycosylated core proteins are essentially the same mass. After reduction, the human 190-kDa HARE gave a single 196-kDa species, which was not seen in the approximately 315-kDa HARE after reduction. The reduced approximately 315-kDa HARE yielded two major proteins at approximately 250 kDa and approximately 220 kDa. We determined the sequence of the human 190-kDa HARE cDNA based on analysis of internal tryptic peptides, as well as RT-PCR and 5' RACE analyses using human spleen and lymph node cDNA libraries. The human gene that encodes HARE is on chromosome 12. PMID- 12626427 TI - Binding inhibition of angiogenic factors by heparan sulfate proteoglycans in aqueous humor: potential mechanism for maintenance of an avascular environment. AB - Aqueous humor is a clear fluid, primarily a blood filtrate, which circulates through the anterior chamber of the eye and bathes the cornea. We explored the possibility that components in the aqueous humor play a direct part in maintaining the avascular environment of the cornea. We report here that heparan sulfate proteoglycan (HSPG) was found in bovine aqueous humor and that it directly inhibits binding of basic fibroblast growth factor and vascular endothelial growth factor to cell-surface heparan sulfate. We demonstrate that this holds true for all heparin binding proteins tested but not for epidermal growth factor, which does not bind heparin. Furthermore, we show, with mathematical modeling, that the concentration of HSPG in aqueous humor (approximately 4 microg/ml), when combined with the clearance of aqueous humor from the eye due to circulation, is sufficient to block the binding of heparin binding growth factors to corneal endothelium. This mechanism suggests a physiological process to control bioavailability of angiogenic growth factors in the cornea. PMID- 12626426 TI - Lovastatin inhibits brain endothelial cell Rho-mediated lymphocyte migration and attenuates experimental autoimmune encephalomyelitis. AB - Neuroinflammatory diseases, such as multiple sclerosis (MS), result from aberrant leukocyte traffic into the central nervous system (CNS). To breach the specialized blood-brain barrier, activated leukocytes interact with CNS endothelial cells (EC) and activate a CD54-mediated signaling pathway controlling the Rho GTPase. To function correctly Rho requires posttranslational prenylation, and this can be inhibited by depleting the supply of isoprenoids through inhibition of the cholesterol synthesis pathway with 3-hydroxy-3-methylglutaryl CoA reductase (HMG-CoA reductase) inhibitors (statins). Here we show that treatment of brain EC in vitro with lovastatin inhibits Rho-mediated transendothelial T cell migration. This effect can be reversed by supplementation with mevalonolactone, the downstream product of HMG-CoA reductase, or by ectopic expression of myristoylated Rho, which remains active in the absence of prenylation. In a relapsing-remitting mouse model of MS, lovastatin treatment inhibited leukocyte migration into the CNS and significantly attenuated the development of both acute and relapsing clinical disease. These studies demonstrate that the indirect pharmacological inhibition of Rho proteins in brain EC by statins can inhibit a key stage in the pathogenesis of neuroinflammation, namely leukocyte migration across the blood-brain barrier. These studies demonstrate a novel effect of statins in modulating the immune response in neuroinflammtory diseases and may provide additional rationale for their use in the treatment of MS. PMID- 12626428 TI - 17beta-estradiol activates ICI 182,780-sensitive estrogen receptors and cyclic GMP-dependent thioredoxin expression for neuroprotection. AB - Clinical studies suggest that estrogen may improve cognition in Alzheimer's patients. Basic experiments demonstrate that 17beta-estradiol protects against neurodegeneration in both cell and animal models. In the present study, a human SH-SY5Y cell model was used to investigate molecular mechanisms underlying the receptor-mediated neuroprotection of physiological concentrations of 17beta estradiol. 17beta-estradiol (<10 nM) concomitantly increased neuronal nitric oxide synthase (NOS1) expression and cell viability. 17beta-estradiol-induced neuroprotection was blocked by the receptor antagonist ICI 182,780, also prevented by inhibitors of NOS1 (7-nitroindazole), guanylyl cyclase (LY 83,583), and cGMP-dependent protein kinase (PKG) (Rp-8-pCPT-cGMPs). In addition to the expression of NOS1 and MnSOD, 17beta-estradiol increased the expression of the redox protein thioredoxin (Trx), which was blocked by the inhibition of either cGMP formation or PKG activity. The expression of heme oxygenase 2 and brain derived neurotrophic factor was not altered. Estrogen receptor-enhanced cell viability against oxidative stress may be linked to Trx expression because the Trx reductase inhibitor, 5,5'-dithio-bis(2-nitrobenzoic acid) significantly reduced the cytoprotective effect of 17beta-estradiol. Furthermore, Trx (1 microM) inhibited lipid peroxidation, proapoptotic caspase-3, and cell death during oxidative stress caused by serum deprivation. We conclude that cGMP dependent expression of Trx--the redox protein with potent antioxidative and antiapoptotic properties--may play a pivotal role in estrogen-induced neuroprotection. PMID- 12626429 TI - Neuroprotective effect of vasoactive intestinal peptide (VIP) in a mouse model of Parkinson's disease by blocking microglial activation. AB - Parkinson's disease (PD) is a common neurodegenerative disorder with no effective protective treatment, characterized by a massive degeneration of dopaminergic neurons in the substantia nigra (SNpc) and the subsequent loss of their projecting nerve fibers in the striatum. To elucidate PD pathogenic factors, and thus to develop therapeutic strategies, a murine PD model based on the administration of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) has been used extensively. It has been demonstrated that activated microglia cells actively participate in the pathogenesis of MPTP-induced PD through the release of cytotoxic factors. Because current treatments for PD are not effective, considerable research focused lately on a number of regulatory molecules termed microglia-deactivating factors. Vasoactive intestinal peptide (VIP), a neuropeptide with a potent anti-inflammatory effect, has been found to be protective in several inflammatory disorders. This study investigates the putative protective effect of VIP in the MPTP model for PD. VIP treatment significantly decreases MPTP-induced dopaminergic neuronal loss in SNpc and nigrostriatal nerve-fiber loss. VIP prevents MPTP-induced activation of microglia in SNpc and striatum and the expression of the cytotoxic mediators, iNOS, interleukin 1beta, and numor necrosis factor alpha. VIP emerges as a potential valuable neuroprotective agent for the treatment of pathologic conditions in the central nervous system, such as PD, where inflammation-induced neurodegeneration occurs. PMID- 12626430 TI - Molecular and functional characterization of SLC26A11, a sodium-independent sulfate transporter from high endothelial venules. AB - Lymphocyte emigration from the blood into most secondary lymphoid organs and chronically inflamed tissues occurs at the level of high endothelial venules (HEV). A unique characteristic of HEV endothelial cells (HEVEC) is their capacity to incorporate large amounts of sulfate into sialomucin-type counter-receptors for the lymphocyte homing receptor L-selectin. We have previously shown that sulfate uptake into HEVEC is mediated by two distinct functional classes of sulfate transporters: Na+-coupled transporters and sulfate/anion exchangers. Here, we report the molecular characterization from human HEVEC of SLC26A11, a novel member of the SLC26 sulfate/anion exchanger family. Functional expression studies in COS-7 and Sf9 insect cells revealed that SLC26A11 is targeted to the cell membrane and exhibits Na+-independent sulfate transport activity, sensitive to the anion exchanger inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). Northern blot analysis showed the highest SLC26A11 transcript levels in placenta, kidney, and brain. The SLC26A11 gene mapped to human chromosome 17q25, very close to the hereditary hearing loss diseases loci DFNA20, DFNA26, and USH1G. RT-PCR analysis of SLC26 sulfate transporters in human HEVEC revealed coexpression of SLC26A11 with SLC26A2/DTDST and lack of SLC26A1/SAT1, SLC26A3/DRA, and SLC26A8/TAT1. Together, our results indicate that SLC26A11 is a novel Na+-independent sulfate transporter that may cooperate with SLC26A2 to mediate DIDS-sensitive sulfate uptake into HEVEC. PMID- 12626431 TI - Besides affecting intracellular calcium signaling, 2-APB reversibly blocks gap junctional coupling in confluent monolayers, thereby allowing measurement of single-cell membrane currents in undissociated cells. AB - 2-aminoethoxydiphenyl borate (2-APB) has been widely used as a blocker of the IP3 receptor and TRP channels, including store-operated calcium channels. We now show in monolayers of normal rat kidney cells (NRK/49F) that 2-APB completely and reversibly blocks gap junctional intercellular communication at concentrations similar to that required for inhibition of PGF2alpha-induced increases in intracellular calcium. Gap junctional conductances between NRK cells were estimated with single-electrode patch-clamp measurements and were fully blocked by 2-APB (50 microM), when applied extracellularly but not via the patch pipette. Half maximal inhibition (IC50) of electrical coupling in NRK cells was achieved at 5.7 microM. Similar results were obtained for human embryonic kidney epithelial cells (HEK293/tsA201) with an IC50 of 10.3 microM. Using 2-APB as an electrical uncoupler of monolayer cells, we could thus measure inward rectifier potassium, L-type calcium, and calcium-dependent chloride membrane currents in confluent NRK monolayers, with properties similar to those in dissociated NRK cells in the absence of 2-APB. The electrical uncoupling action described here is a new 2-APB property that promises to provide a powerful pharmacological tool to study single-cell properties in cultured confluent monolayers and intact tissues by electrical and chemical uncoupling of the cells without the need of prior dissociation. PMID- 12626432 TI - Glycation proceeds faster in mutated Cu, Zn-superoxide dismutases related to familial amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) involves the progressive degeneration of motor neurons in the spinal cord and motor cortex. It has been shown that 15-20% of patients with familial ALS (FALS) have defects in the Sod1 gene that encodes Cu, Zn-superoxide dismutase (SOD). To elucidate the pathological role of mutated Cu, Zn-SODs in FALS, the susceptibility of mutants to glycation was examined. Mutated Cu, Zn-SODs (G37R, G93A, and I113T) related to FALS and wild type were produced in a baculovirus/insect cell expression system. Glycated and nonglycated proteins were separated on a boronate column, and the nonglycated fraction was then incubated with glucose. The mutated Cu, Zn-SODs were found to be highly susceptible to glycation compared with the wild-type enzyme as estimated by Western blot analysis using an anti-hexitol lysine antibody. The mutated Cu, Zn SOD incubated with glucose generated higher levels of hydrogen peroxide than the wild-type enzyme. Mutated Cu, Zn-SODs were also shown to be highly susceptible to fructation, and the fructated mutant also produced higher levels of hydrogen peroxide than the wild type. These results suggest that high susceptibility of mutated Cu, Zn-SODs to glycation could be the origin of the oxidative stress associated with neuronal dysfunction in FALS. PMID- 12626433 TI - Gelatinase B is diabetogenic in acute and chronic pancreatitis by cleaving insulin. AB - Genetic, endocrine, and environmental factors contribute to the development of diabetes. Much information has been gathered on the homeostasis mechanisms of glucose regulation by insulin-producing pancreatic beta cells. Here we demonstrate high expression levels of gelatinase B (matrix metalloproteinase-9, MMP-9) by neutrophils in acute pancreatitis and by ductular epithelial cells in chronic pancreatitis. Because gelatinase B processes cytokines and chemokines, we investigated whether and how gelatinase B cleaves insulin. Pure human neutrophil gelatinase B was found to destroy insulin by cleavage at 10 sites. Pancreatic islet and ductular cells are relatively spared in comparison with the complete destruction of acinar cells of the exocrine pancreas in chronic pancreatitis. High expression levels of gelatinase B are maintained in the immediate proximity of insulin-secreting beta cells. Consequently, diabetes may be worsened by enzymatic degradation of insulin by gelatinase B and by the consequent enhancement of the autoimmune process. Gelatinase B is diabetogenic in acute and chronic pancreatitis by cleaving insulin. PMID- 12626434 TI - cDNA gene expression profile homology of antioxidants and their antiapoptotic and proapoptotic activities in human neuroblastoma cells. AB - Antioxidants have concentration-dependent neuroprotective and proapoptotic activities in models of Parkinson's disease. The aim of our study was to determine gene-protein pathways of the antioxidants, dopamine (DA), R-apomorphine (R-APO), melatonin, and green tea polyphenol (-)-epigallocatechin-3-gallate (EGCG), in neuroblastoma cells, using a customized cDNA microarray and quantitative reverse transcriptase-polymerase chain reaction gene expression techniques. We demonstrate a concentration-dependent correlation between these compounds and modulation of cell survival/cell death-related gene pathways. High toxic concentration of DA (500 microM), R-APO (50 microM), melatonin (50 microM), and EGCG (50 microM) exhibited a similar profile of proapoptotic gene expression, increasing the level of bax, caspase-6, fas ligand, and the cell-cycle inhibitor gadd45 genes, while decreasing antiapoptotic bcl-2 and bcl-xL. Conversely, the low neuroprotective concentrations (1-10 microM) of these compounds induced an antiapoptotic response. Melatonin displayed an extremely low index of mortality, which may be partially explained by the observation that a high concentration did not significantly affect the expression of mitochondrial Bcl-2 family members, bcl-2 and bax. Protein analysis of Bcl-2, Bax, and activated caspase-3 correlated with the gene expression pattern. Our results provide for the first time new insights into the molecular events involved in the dose-dependent neuroprotective and neurotoxic activities of catechols and indole amine compounds. PMID- 12626435 TI - Microarray analysis of nonhuman primates: validation of experimental models in neurological disorders. AB - Nonhuman primates (NHPs) have provided robust experimental animal models for many human-related diseases due to their similar physiologies. Nonetheless, profound differences remain in the acquisition, progression, and outcome of important diseases such as AIDS and Alzheimer's, for which the underlying basis remains obscure. We explored the utility of human high-density oligonucleotide arrays to survey the transcription profile of NHP genomes. Total RNA from prefrontal cortices of human (Homo sapiens), common chimpanzee (Pan troglodytes), cynomolgous macaque (Macaca fascicularis), and common marmoset (Callithrix jacchus) was labeled and hybridized to Affymetrix U95A GeneChip probe arrays. Corresponding data obtained previously from common chimpanzee and orangutan (Pongo pygmaeus) were added for comparison. Qualitative (present or not detected) and quantitative (expression level) analysis indicated that many genes known to be involved in human neurological disorders were present and regulated in NHPs. A gene involved in dopamine metabolism (catechol-O-methyltransferase) was absent in macaque and marmoset. Glutamate receptor 2 was up-regulated, and transcription associated genes were down-regulated in NHPs compared with humans. We demonstrate that transcript profiling of NHPs could provide comparative genomic data to validate and better focus experimental animal models of human neurological disorders. PMID- 12626436 TI - Exercise and IL-6 infusion inhibit endotoxin-induced TNF-alpha production in humans. AB - During "nondamaging" exercise, skeletal muscle markedly releases interleukin (IL) 6, and it has been suggested that one biological role of this phenomenon is to inhibit the production of tumor necrosis factor (TNF)- alpha, which is known to cause pathogenesis such as insulin resistance and atherosclerosis. To test this hypothesis, we performed three experiments in which eight healthy males either rested (CON), rode a bicycle for 3 h (EX), or were infused with recombinant human IL-6 (rhIL-6) for 3 h while they rested. After 2.5 h, the volunteers received a bolus of Escherichia coli lipopolysaccharide endotoxin (0.06 ng/kg) i.v. to induce low-grade inflammation. In CON, plasma TNF-alpha increased significantly in response to endotoxin. In contrast, during EX, which resulted in elevated IL 6, and rhIL-6, the endotoxin-induced increase in TNF-alpha was totally attenuated. In conclusion, physical exercise and rhIL-6 infusion at physiological concentrations inhibit endotoxin-induced TNF-alpha production in humans. Hence, these data provide the first experimental evidence that physical activity mediates antiinflammatory activity and suggest that the mechanism include IL-6, which is produced by and released from exercising muscles. PMID- 12626437 TI - Regulation of the intestinal epithelial response to cyclic strain by extracellular matrix proteins. AB - Repetitive mechanical deformation may stimulate intestinal epithelial proliferation. Because the extracellular matrix modulates static intestinal epithelial biology, we examined whether matrix proteins influence intestinal epithelial responses to deformation. Human Caco-2BBE cells and nontransformed human enterocytes (HIPEC) were subjected to 10% average cyclic strain at 10 cycles/min on flexible membranes precoated with matrix proteins without or with plasma fibronectin or functional anti-integrin antibodies in the medium. Strain stimulated proliferation, focal adhesion kinase, extracellular signal-regulated protein kinase (ERK), p38, and Jun N-terminal kinase similarly on collagen I or IV, and more weakly on laminin, but had no effect on fibronectin. MEK blockade (PD98059) prevented strain-stimulated proliferation on collagen but did not affect proliferation on fibronectin. Adding tissue fibronectin to a collagen substrate or plasma fibronectin to the media suppressed strain s mitogenic and signal effects, but not those of epidermal growth factor. Functional antibodies to the alpha5 or alpha(v) integrin subunit blocked strain's effects on Caco-2 proliferation and ERK activation, although ligation of the alpha2 or alpha6 subunit did not. Repetitive strain also stimulated, and fibronectin inhibited, human intestinal primary epithelial cell proliferation. Repetitive deformation stimulates transformed and nontransformed human intestinal epithelial proliferation in a matrix-dependent manner. Tissue or plasma fibronectin may regulate the intestinal epithelial response to strain via integrins containing alpha5 or alpha(v). PMID- 12626438 TI - Hypoxia causes down-regulation of 11 beta-hydroxysteroid dehydrogenase type 2 by induction of Egr-1. AB - Hypoxia causes several renal tubular dysfunctions, including abnormal handling of potassium and sodium and increased blood pressure. Therefore, we investigated the impact of hypoxia on 11beta-hydroxysteroid dehydrogenase (11beta-HSD2) enzyme, a crucial prereceptor gatekeeper for renal glucocorticosteroid-mediated mineralocorticoid action. The effect of hypoxia was assessed in vitro by incubating LLC-PK1 cells with antimycin A, an inhibitor of mitochondrial oxidative phosphorylation. Antimycin A induced a dose- and time-dependent reduction of 11beta-HSD2 activity. The early growth response gene, Egr-1, a gene known to be stimulated by hypoxia was investigated because of a potential Egr-1 binding site in the promoter region of 11beta-HSD2. Antimycin A induced Egr-1 protein and Egr-1-regulated luciferase gene expression. This induction was prevented with the MAPKK inhibitor PD 98059. Overexpression of Egr-1 reduced endogenous 11beta-HSD2 activity in LLC-PK1 cells, indicating that MAPK ERK is involved in the regulation of 11beta-HSD2 in vitro. In vivo experiments in rats revealed that Egr-1 protein increases, whereas 11beta-HSD2 mRNA decreases, in kidney tissue after unilateral renal ischemia and in humans the renal activity of 11beta-HSD2 as assessed by the urinary ratio of (tetrahydrocortisol+5alpha tetrahydrocortisol)/tetrahydrocortisone declined when volunteers were exposed to hypoxemia at high altitude up to 7000 m. Thus, hypoxia decreases 11beta-HSD2 transcription and activity by inducing Egr-1 in vivo and in vitro. This mechanism might account for enhanced renal sodium retention and hypertension associated with hypoxic conditions. PMID- 12626439 TI - Phenotypic knock out of heparan sulfates in myotubes impairs excitation-induced calcium spiking. AB - Little is known about the physiological functions of heparan sulfates (HSs), which are present in the basal lamina surrounding skeletal muscle fibers. Here, we present a new system in which HS is phenotypically knocked out by endogenous expression of epitope-specific anti-HS antibodies. Single-chain antibodies, containing an immunoglobulin leader peptide, were produced by using various expression systems. Antibodies were detected in the Golgi apparatus, the site of HS biosynthesis. Likewise, the HS-degrading enzyme heparanase was expressed. Endogenous expression of antibodies or heparanase in myoblasts resulted in HS defective myotubes. Excitability and calcium kinetics of HS-defective myotubes were severely compromised, as determined by analysis of electrically induced calcium spikes via video-speed UV confocal laser scanning microscopy. Phenotypically knocking out of individual HS epitopes resulted in specific effects on excitability and calcium kinetics. These data indicate important roles for HSs in skeletal muscle calcium kinetics. PMID- 12626440 TI - Laminin-1 peptide-conjugated chitosan membranes as a novel approach for cell engineering. AB - Laminin, a major component of the basement membrane, has diverse biological activities. Recently, we identified various biologically active sequences on laminin-1 by using a large set of synthetic peptides. Chitosan, a polysaccharide, is biodegradable and has been used as a biomaterial. Here, we conjugated several biologically active laminin peptides onto chitosan membranes and measured the cell attachment activity of peptide-conjugated chitosan membranes with various cell types. The active laminin peptide-conjugated chitosan membranes promoted cell attachment with cell type specificity. A99 (AGTFALRGDNPQG)-chitosan membrane promoted cell attachment with well-organized actin stress fibers. This adhesion was inhibited by EDTA but not by heparin. AG73 (RKRLQVQLSIRT)-chitosan membrane promoted cell attachment with filopodia formation, and this adhesion was inhibited by heparin but not by EDTA. These data suggest that the A99-chitosan membrane interacted with an integrin cellular receptor and that the AG73-chitosan membrane promoted proteoglycan-mediated cell attachment, as previously reported. Furthermore, both AG73-chitosan and A99-chitosan membranes effectively promoted neurite outgrowth with PC12 rat pheochromocytoma cells. We conclude that conjugation on a chitosan membrane is applicable for testing quantitatively the biological activity of synthetic peptides and that these constructs have a potential ability to serve as bioadhesive materials for tissue regeneration and engineering. PMID- 12626441 TI - Long-lasting effects of neonatal dexamethasone treatment on spatial learning and hippocampal synaptic plasticity: involvement of the NMDA receptor complex. AB - The effects of neonatal dexamethasone (DEX) treatment on spatial learning and hippocampal synaptic plasticity were investigated in adult rats. Spatial learning in reference and working memory versions of the Morris maze was impaired in DEX treated rats. In hippocampal slices of DEX rats, long-term depression was facilitated and potentiation was impaired. Paired-pulse facilitation was normal, suggesting a postsynaptic defect as cause of the learning and plasticity deficits. Western blot analysis of hippocampal postsynaptic densities (PSD) revealed a reduction in NR2B subunit protein, whereas the abundance of the other major N-methyl-D-aspartate (NMDA) receptor subunits (NR1, NR2A), AMPA receptor subunits (GluR2/3), scaffolding proteins, and Ca2+/calmodulin-dependent protein kinase II (alphaCaMKII) were unaltered. This selective reduction in NR2B likely resulted from altered receptor assembly rather than subunit expression, because the abundance of NR2B in the homogenate and crude synaptosomal fractions was unaltered. In addition, the activity of alphaCaMKII, an NMDA receptor complex associated protein kinase, was increased in PSD of DEX rats. The results indicate that neonatal treatment with DEX causes alterations in composition and function of the hippocampal NMDA receptor complex that persist into adulthood. These alterations likely explain the deficits in hippocampal synaptic plasticity and spatial learning induced by neonatal DEX treatment. PMID- 12626442 TI - Total C4B deficiency due to gene deletion and gene conversion in a patient with severe infections. AB - Deficiencies of the early components of the classical complement pathway impair the actions of innate and humoral immunity and may lead to increased susceptibility to infections. We have studied the genetic basis of total C4B deficiency in a Finnish patient with recurrent meningitis, chronic fistulas and abscesses. The maternal chromosome carried a four-gene deletion including the C4B gene, and a conversion from C4B to C4A gene was found on the paternal chromosome resulting in complete deficiency of C4B. In the converted C4A gene, mutation screening did not reveal any amino acid changes or prominent mutations, yet a large number of nucleotide variations were found. Further, the patient was heterozygous for structural deficiency of mannan binding lectin (MBL) associating with medium levels of serum MBL. Our data provides new information on the genetic instability of the C4 gene region, and on the association of homozygous C4B deficiency and variant MBL genotype with increased susceptibility to recurrent and chronic infections. Importantly, plasma therapy induced a prompt clinical cure with long-term effects. PMID- 12626443 TI - Levels of antibodies specific to tetanus toxoid, Haemophilus influenzae type b, and pneumococcal capsular polysaccharide in healthy children and adults. AB - Antibody levels specific for capsular polysaccharides of Streptococcus pneumoniae and Haemophilus influenzae type b (Hib) and for tetanus toxoid were measured in serum samples of 386 age-stratified subjects. The study group consists of healthy adult blood donors and hospitalized children undergoing elective surgery, excluding individuals with a history of infection. In children, anti-tetanus toxoid antibody levels displayed two peaks of 1.20 IU/ml (20.4 mg/liter) and 1.65 IU/ml (28.1 mg/liter) related to the schedule of routine childhood immunization in the first year and at 8 years of age. Eighty percent of the antibodies are of the immunoglobulin G1 (IgG1) isotype. For pneumococcal capsular polysaccharide (PCP), the specific antibody levels represent the acquisition of natural immunity. The initial concentration of 9.2 mg/liter was low in infancy (0.5 to 1 years of age) and remained low until 3 to 4 years of age (14.6 mg/liter). During this period PCP antibodies were almost 100% of the IgG2 subclass. Thereafter, IgG anti-PCP antibody titers increased steadily to adult levels (59.5 mg/liter). The data are intended to provide reference ranges to aid in the interpretation of specific antibody determinations in the clinical setting. PMID- 12626444 TI - Clearance of virulent but not avirulent Rhodococcus equi from the lungs of adult horses is associated with intracytoplasmic gamma interferon production by CD4+ and CD8+ T lymphocytes. AB - Rhodococcus equi is a gram-positive bacterium that infects alveolar macrophages and causes rhodococcal pneumonia in horses and humans. The virulence plasmid of R. equi appears to be required for both pathogenicity in the horse and the induction of protective immunity. An understanding of the mechanisms by which virulent R. equi circumvents protective host responses and by which bacteria are ultimately cleared is important for development of an effective vaccine. Six adult horses were challenged with either virulent R. equi or an avirulent, plasmid-cured derivative. By using a flow cytometric method for intracytoplasmic detection of gamma interferon (IFN-gamma) in equine bronchoalveolar lavage fluid (BALF) cells, clearance of the virulent strain was shown to be associated with increased numbers of pulmonary CD4(+) and CD8(+) T lymphocytes producing IFN gamma. There was no change in IFN-gamma-positive cells in peripheral blood, suggesting that a type 1 recall response at the site of challenge was protective. The plasmid-cured strain of R. equi was cleared in horses without a significant increase in IFN-gamma-producing T lymphocytes in BALF. In contrast to these data, a previous report in foals suggested an immunomodulating role for R. equi virulence plasmid-encoded products in downregulating IFN-gamma expression by equine CD4(+) T lymphocytes. Intracytoplasmic detection of IFN-gamma provides a method to better determine whether modulation of macrophage-activating cytokines by virulent strains occurs uniquely in neonates and contributes to their susceptibility to rhodococcal pneumonia. PMID- 12626445 TI - Simple method to distinguish between primary and secondary C3 deficiencies. AB - Due to the increasing numbers of reported clinical cases of complement deficiency in medical centers, clinicians are now more aware of the role of the complement system in the protection against infections caused by microorganisms. Therefore, clinical laboratories are now prepared to perform a number of diagnostic tests of the complement system other than the standard 50% hemolytic component assay. Deficiencies of alternative complement pathway proteins are related to severe and recurrent infections; and the application of easy, reliable, and low-cost methods for their detection and distinction are always welcome, notably in developing countries. When activation of the alternative complement pathway is evaluated in hemolytic agarose plates, some but not all human sera cross-react to form a late linear lysis. Since the formation of this linear lysis is dependent on C3 and factor B, it is possible to use late linear lysis to routinely screen for the presence of deficiencies of alternative human complement pathway proteins such as factor B. Furthermore, since linear lysis is observed between normal human serum and primary C3-deficient serum but not between normal human serum and secondary C3-deficient serum caused by the lack of factor H or factor I, this assay may also be used to discriminate between primary and secondary C3 deficiencies. PMID- 12626446 TI - Antigenic classification of Rickettsia felis by using monoclonal and polyclonal antibodies. AB - Rickettsia felis is a flea-transmitted rickettsia. There is a discrepancy between its reported phylogenic and phenotypic identifications. Following the first report of R. felis, it was considered by tests with serologic reagents to be closely related to another recognized flea-transmitted rickettia, R. typhi. Subsequently, it appeared to be more closely related to spotted fever group (SFG) rickettsiae by genetic analysis. In the present work, R. felis was studied by microimmunofluorescence (MIF) serologic typing and with monoclonal antibodies (MAbs). Mouse polyclonal antisera to R. felis cross-reacted only with SFG rickettsiae. A neighbor-joining analysis based on MIF indicated that R. felis is actually related to SFG rickettsiae antigenically, clustering with R. australis, R. akari, and R. montanensis. A panel of 21 MAbs was raised against a 120-kDa protein antigen or a 17-kDa polypeptide of R. felis. They cross-reacted with most members of the SFG rickettsiae but not with R. prowazekii, R. typhi, or R. canadensis of the typhus group (TG) rickettsiae. Sixty-four MAbs previously generated to seven other ricketttsial species were tested with R. felis. Three MAbs reacted with the 120-kDa antigen and were generated by R. africae, R. conorii, and R. akari, respectively. They exhibited cross-reactivities with R. felis. All our data show that R. felis harbors the antigenic profile of an SFG rickettsia. PMID- 12626447 TI - Detection of cross-reactivity for atopic immunoglobulin E against multiple allergens. AB - The existence of specific immunoglobulin E (IgE) allows us to determine the allergens that cause the allergic disease. For the purposes of allergen avoidance and immunotherapy, the measurement of specific IgE is widely applied in clinical laboratories. However, if IgE from the serum of an allergic patient exhibits reactivity to multiple allergens, it would cause a problem. The present study analyzes whether the serum IgE with multiple reactivity is due to the presence of unique IgE against the common epitope shared by different allergens or the presence of multiple IgEs against different epitopes on different allergens. The quantitative-competitive inhibition tests and the immunoblotting were applied to analyze the immunosimilarity among examined allergens. The result shows that the competitive inhibition of IgE binding between shrimp and crab allergens is higher than those between either shrimp and cockroach or between crab and cockroach. Furthermore, the results of immunoblotting are consistent with those of quantitative-competitive inhibition tests. These results allow us to detect the cross-reactivity for atopic IgE against multiple allergens. PMID- 12626449 TI - Performance of five serological assays for diagnosis of Rhodococcus equi pneumonia in foals. AB - The performance of four enzyme-linked immunosorbent assays (ELISAs) (ELISA-6939, ELISA-33701, ELISA-VapA, and ELISA-California) and an agar gel immunodiffusion test for diagnosis of Rhodococcus equi pneumonia in foals was evaluated. Antibody concentrations of foals with culture-confirmed R. equi pneumonia (n = 41) were compared to those of age-matched pasturemates that remained clinically healthy during the entire breeding season (n = 24). For each serological assay evaluated, selection of a low cutoff resulted in high sensitivity but low specificity. Increasing the cutoff value resulted in better specificity but to the detriment of sensitivity. The best diagnostic performance was achieved with ELISA California at a cutoff of 40%, resulting in a sensitivity of 59% and a specificity of 88%. We conclude that current serological assays do not differentiate between diseased and clinically healthy foals. PMID- 12626448 TI - Rapid detection of contagious bovine pleuropneumonia by a Mycoplasma mycoides subsp. mycoides SC capsular polysaccharide-specific antigen detection latex agglutination test. AB - A latex agglutination test (LAT) has been developed for the diagnosis of contagious bovine pleuropneumonia (CBPP). The latex microspheres were coated with MmmSC polyclonal immunoglobulin G antiserum and detected MmmSC antigen in the serum of cattle infected with CBPP and in growth medium containing MmmSC. The specific antigen recognized by this test appeared to be the capsular polysaccharide (CPS). The LAT recognized all 23 strains of MmmSC examined in this study, with a sensitivity level of 2 ng of CPS, or the equivalent of 5 x 10(3) CFU, in a reaction volume of 0.03 ml. Therefore, rapid identification of MmmSC cultures should be possible. Agglutination was also observed with the related goat pathogens and "Mycoplasma mycoides" cluster members Mycoplasma mycoides subsp. mycoides large colony biotype (four of six strains positive) and Mycoplasma mycoides subsp. capri (three of six strains positive), in agreement with the suggestion that these latter two mycoplasmas may in fact represent a single species (although collectively exhibiting two capsular serotypes). Comparisons in diagnosis with the complement fixation test (CFT) were made by using African field sera from CBPP-infected cattle. After 2 (or 3) min of incubation, the test detected 55% (or 61%) of CFT-positive sera and 29% (or 40%) of CFT-negative sera, with an overall correlation in diagnosis of 62% (or 61%). The rates for false-positive diagnoses made by using "known" CBPP-negative sera from the United Kingdom were 3 or 13% after 2 or 3 min of incubation, respectively. The data agree with previous findings that some CBPP CFT-negative misdiagnoses may occur due to "antibody eclipsing" by excess circulating antigen. The LAT combines low cost and high specificity with ease of application in the field, without the need for any specialist training or equipment. PMID- 12626450 TI - Inhibition of pneumococcal adherence to human nasopharyngeal epithelial cells by anti-PsaA antibodies. AB - The role of pneumococcal (Pnc) surface adhesin A (PsaA) in the adherence of Streptococcus pneumoniae (pneumococcus) to host cells is not well defined. We examined the effect of anti-PsaA antibodies in an inhibition of adherence assay using Detroit 562 nasopharyngeal human epithelial cells. Rabbit polyclonal (Pab) anti-recombinant PsaA (rPsaA) sera, a purified mouse monoclonal antibody (MAb) (MAb 6F62G8E12), and 22 healthy adult sera with known anti-PsaA IgG levels (obtained by enzyme-linked immunosorbent assay) were evaluated for their abilities to inhibit Pnc adherence to confluent monolayers (measured as percent reduction in CFU counts compared to those of uninhibited controls). Pnc adherence was dependent on capsular phenotype (no or low adherence for opaque strains). With an inoculum of 10(4) to 10(5) bacteria/well, the mean +/- standard deviation count in controls was 163 +/- 32 CFU/well for transparent strains. Low adherence was observed for a PsaA-minus mutant even at higher inoculum doses. Mean percent inhibitions of adherence with Pab and MAb were 54 and 50%, respectively. Adult sera showed inhibition in a dose-response fashion with a range of 98 to 8%, depending on the serum anti-PsaA antibody concentration. Absorption of Pab with rPsaA restored Pnc adherence to control levels. Absorption of sera with a PsaA minus mutant did not result in a significant decrease (P >0.05) of inhibition of adherence activity. Additionally, nearly 100% of Pnc adherence was inhibited by lipidated rPsaA at 2.5 micro g/ml. Our data support the argument that PsaA is an adhesin that mediates Pnc adherence to human nasopharyngeal cells. This functional assay may be useful in evaluating antibodies elicited in response to PsaA vaccination. PMID- 12626451 TI - Contribution of epitope specificity to the binding of monoclonal antibodies to the capsule of Cryptococcus neoformans and the soluble form of its major polysaccharide, glucuronoxylomannan. AB - Incubation of encapsulated cryptococci with monoclonal antibodies (MAbs) specific for glucuronoxylomannan (GXM), the major capsular polysaccharide of Cryptococcus neoformans, produces two distinct capsular quellung-type reactions termed rim and puffy. The type of capsular reaction that occurs is determined by the epitope specificity of the MAb and the serotype of the yeast cell. Several biological activities, including opsonic activity, complement activation, and protective efficacy, are associated with the type of capsular reaction produced by a MAb. The goal of this study was to examine the reactivities of two families of anti GXM MAbs with serotype A and D capsular polysaccharides in several immunochemical assays, including agglutination, immunofluorescence, quantitative precipitation, and enzyme-linked immunosorbent assay, in an effort to identify serological assays that are predictive of the capsular quellung reaction. The results showed that the type of capsular reaction (rim versus puffy) is a qualitative assessment of antibody-capsule interaction that cannot be predicted on the basis of a serological assay. The results further showed that antibody reactivity demonstrated in one serological assay is not necessarily predictive of results in another assay, particularly in cases where one assay examines antibody-capsule interactions, e.g., agglutination, and another assay examines interaction of antibody with soluble GXM. Taken together, the results suggest caution in interpretation of immunochemical assays for anti-GXM antibodies and recommend the use of multiple assays formats when studying anticryptococcal antibodies. PMID- 12626452 TI - Lipoteichoic acids from Lactobacillus strains elicit strong tumor necrosis factor alpha-inducing activities in macrophages through Toll-like receptor 2. AB - Lactobacilli are nonpathogenic gram-positive inhabitants of microflora. At least some Lactobacillus strains have been postulated to have health beneficial effects, such as the stimulation of the immune system. Here we examined the stimulatory effects of lactobacilli on mouse immune cells. All six heat-killed Lactobacillus strains examined induced the secretion of tumor necrosis factor alpha (TNF-alpha) from mouse splenic mononuclear cells, albeit to various degrees. When fractionated subcellular fractions of Lactobacillus casei were tested for NF-kappaB activation and TNF-alpha production in RAW264.7, a mouse macrophage cell line, the activity was found to be as follows: protoplast > cell wall >> polysaccharide-peptidoglycan complex. Both crude extracts and purified lipoteichoic acids (LTAs) from two Lactobacillus strains, L. casei and L. fermentum, significantly induced TNF-alpha secretion from RAW264.7 cells and splenocytes of C57BL/6, C3H/HeN, and C3H/HeJ mice but not from splenocytes of C57BL/6 TLR2(-/-) mice. Lactobacillus LTA induced activation of c-Jun N-terminal kinase activation in RAW264.7 cells. Furthermore, in HEK293T cells transected with a combination of CD14 and Toll-like receptor 2 (TLR2), NF-kappaB was activated in response to Lactobacillus LTA. Taken together, these data suggest that LTAs from lactobacilli elicit proinflammatory activities through TLR2. PMID- 12626453 TI - Competitive-inhibition enzyme-linked immunosorbent assay for detection of serum antibodies to caprine arthritis-encephalitis virus: diagnostic tool for successful eradication. AB - A competitive-inhibition enzyme-linked immunosorbent assay (cELISA) was evaluated for the detection of serum antibodies to the surface envelope (SU) of caprine arthritis-encephalitis virus (CAEV) in goats. This assay utilized 96-well microtiter plates containing CAEV-63 SU captured by monoclonal antibody (MAb) F7 299 and measured the competitive displacement of horseradish peroxidase conjugated MAb GPB 74A binding by undiluted goat sera (F. Ozyoruk, W. P. Cheevers, G. A. Hullinger, T. C. McGuire, M. Hutton, and D. P. Knowles, Clin. Diagn. Lab. Immunol. 8:44-51, 2001). Two hundred serum samples from goats in the United States were used to determine the sensitivity and specificity of cELISA based on the immunoprecipitation (IP) of [(35)S]methionine-labeled viral antigens as a standard of comparison. A positive cELISA was defined as >33.2% inhibition of MAb 74A binding based on 2 standard deviations above the mean percent inhibition of 140 IP-negative serum samples. At this cutoff value, there were 0 of 60 false-negative sera (100% sensitivity) and 5 of 140 false-positive sera (96.4% specificity). Additional studies utilized IP-monitored cELISA to establish a CAEV-free herd of 1,640 dairy goats. PMID- 12626454 TI - Leucotoxic activities of Staphylococcus aureus strains isolated from cows, ewes, and goats with mastitis: importance of LukM/LukF'-PV leukotoxin. AB - Among the toxins that Staphylococcus aureus is able to secrete, bi-component toxins named leukotoxins target specifically leukocytes, mainly phagocytic cells. Isolates from cows, goats and ewes with mastitis were selected on the basis of the presence or not of the genes encoding the recently described LukM/LukF'-PV leukotoxin. Of the 128 isolates tested, 126 had moderate to high leukotoxic activity to bovine polymorphonuclear cells (PMN). The supernatants of lukM positive isolates were much more leukotoxic than the supernatants of lukM negative isolates: mean leukotoxic titers were 122 versus 20 and 581 versus 26 for isolates of bovine and caprine origin, respectively. Among lukM/lukF'-PV positive isolates, those of caprine and ovine origins were more leukotoxic than were isolates of bovine origin (P < 0.01). The two most abundant proteins in the culture supernatant of a highly toxic isolate were purified and identified as the two components of LukM (LukM and LukF'-PV) on the basis of their molecular mass, N-terminal amino acid sequence, and high synergistic activity. LukM/LukF'-PV induced the flattening of bovine PMN at a concentration as low as 3.6 ng/ml (0.1 nM). A higher concentration (18 ng/ml) was necessary to produce the same effect on caprine or ovine PMN. Affinity-purified antibodies to LukM or to LukF'-PV neutralized the leukotoxic effect of all the culture supernatants. They neutralized with the same efficiency the toxic activity of supernatants from lukM/lukF'-PV positive or negative isolates. These results establish that LukM/LukF'-PV is very active on PMN of ruminants and suggest that this leukotoxin could be the most active leukotoxin produced by mastitis isolates. They prompt further studies to delineate the contribution of LukM/LukF'-PV to the pathogenesis of mastitis in ruminants and the protective effect of antibodies to this leukotoxin. PMID- 12626455 TI - Serological cross-reactivities between antibodies to simian virus 40, BK virus, and JC virus assessed by virus-like-particle-based enzyme immunoassays. AB - Enzyme immunoassays (EIAs) for detection of serum antibodies to simian virus 40 (SV40), BK virus (BKV), and JC virus (JCV) were developed by using virus-like particles (VLPs) produced in insect cells from recombinant baculoviruses expressing the VP1 protein of the respective virus. Rhesus macaque sera with neutralizing antibodies to SV40 showed a high level of reactivity in the SV40 VLP based EIA, and these sera also showed lower levels of reactivity in the BKV and JCV VLP-based EIAs. Rhesus macaque sera negative for neutralizing antibodies to SV40 were negative in all three EIAs. Competitive binding assays showed that SV40 VLPs inhibited BKV reactivity. In rhesus macaque sera, high optical density (OD) values for antibodies to SV40 VLPs were correlated with high OD values for antibodies to BKV but not with high OD values for antibodies to JCV VLPs. Human sera with neutralizing antibodies to SV40 were more reactive to SV40 VLPs than human sera without neutralizing antibodies to SV40. The greater SV40 reactivities of human sera were correlated with greater reactivities to BKV VLPs but not JCV VLPs. These data suggest that cross-reactivity with BKV antibodies may account for part of the low-level SV40 reactivity seen in human sera. With their greater versatility and their suitability for large-scale testing, the VLP-based EIAs for SV40, BKV, and JCV are likely to contribute to a better understanding of the biology of these viruses. PMID- 12626456 TI - Constitutive expression of a bacterial pattern recognition receptor, CD14, in human salivary glands and secretion as a soluble form in saliva. AB - Saliva contains a number of proteins and glycoproteins that protect oral tissues, but little is known about the role of human saliva in innate immunity. Here we showed that human major salivary gland cells constitutively expressed a bacterial pattern recognition receptor, CD14, by immunohistochemistry. Human salivary gland cells in culture express CD14 mRNA and a 55-kDa CD14 protein in, but not on the cells, and secrete a soluble form with the same molecular mass. Human whole saliva contains a 55-kDa CD14, and the concentration of parotid saliva was 10 fold higher than whole saliva, which is comparable to that of serum CD14. Levels of CD14 in unstimulated whole and parotid saliva were unchanged before and after a meal and between unstimulated and stimulated saliva, indicating that saliva CD14 is constitutively secreted into the oral cavity. In contrast, lipopolysaccharide (LPS)-binding protein was below the detectable level. The saliva CD14 is functionally active in that it mediated the activation of CD14 lacking intestinal epithelial cells by LPS in a Toll-like receptor 4-dependent manner. These results suggested that saliva CD14 is important for the maintenance of oral health and possibly intestinal homeostasis. PMID- 12626457 TI - Correlation of organism burden and alveolar macrophage counts during infection with Pneumocystis carinii and recovery. AB - Changes in the number of alveolar macrophages were correlated with organism burden during Pneumocystis carinii infection. The lungs of healthy, dexamethasone treated, and dexamethasone-treated and P. carinii-infected rats were lavaged with phosphate-buffered saline. Counting of alveolar macrophages in the lavage fluids revealed that P. carinii infection caused a 58% decrease in the number of alveolar macrophages and that higher P. carinii organism burdens caused a more rapid decrease in alveolar macrophage number. As a control, healthy rats were challenged with the same number of organisms as that normally used to generate P. carinii infections in dexamethasone-treated rats. Thirteen days after challenge, these rats had a profound (54%) increase in alveolar macrophage number in response to the challenge, while the number of alveolar macrophages in immunosuppressed and P. carinii-infected rats had decreased significantly by this time point. These experiments created the first animal model to mimic human pneumocystis pneumonia in alveolar macrophage number alterations. Reduction of P. carinii organism numbers by treatment of rats with trimethoprim and sulfamethoxazole brought a slow rebound in alveolar macrophage number, while recovery from P. carinii infection by cessation of immunosuppression brought a rapid rebound in alveolar macrophage number. These results suggest that both the immune state of the host and P. carinii burden affect alveolar macrophage number. PMID- 12626458 TI - Performance characteristics of a rapid new immunochromatographic test for detection of antibodies to human immunodeficiency virus. AB - A new immunochromatographic rapid test (Rapid Check HIV 1 and 2; Nucleo de Doencas Infecciosas) for the detection of antibodies to human immunodeficiency virus type 1 and type 2 in human samples (whole blood, serum, and plasma) was evaluated and compared to the commercially available Determine (Abbott Laboratories). When whole-blood samples were evaluated, the specificity and sensitivity of both tests were 100%. However, when plasma samples were used, sensitivity for the Rapid Check HIV 1&2 and the Determine tests were 100 and 98.58%, respectively. The observed specificity for plasma samples was 98.94% for the Rapid Check HIV 1&2 and 96.97% for the Determine test. The results presented here are encouraging and support the adoption of both tests as an alternative to enzyme-lined immunosorbent assay and/or Western blots in regions where laboratorial infrastructure is not available or for use in the management of occupational accidents for healthcare workers. PMID- 12626460 TI - RNase L levels in peripheral blood mononuclear cells: 37-kilodalton/83-kilodalton isoform ratio is a potential test for chronic fatigue syndrome. AB - Chronic fatigue syndrome (CFS) is a disorder characterized by debilitating fatigue associated with immunological abnormalities. The etiology remains unclear. A low-molecular-mass (37 kDa) isoform of RNase L has been described in peripheral blood mononuclear cell (PBMC) extracts, and the ratio of two isoforms of RNase L (37 kDa/83 kDa) has been proposed as a potential biochemical marker of CFS. In a prospective case-control study, we tested whether the RNase L 37-kDa/83 kDa ratio could discriminate a SFC population. We compared the ratio of RNase L isoforms in PBMCs from 11 patients with CFS (6 women and 5 men; mean age +/- standard deviation, 43.2 +/- 13.8 years) and PBMCs from 14 healthy well-matched volunteers (10 women and 4 men; age, 39.1 +/- 11.6 years). A ratio of RNase L of 0.4 used as a threshold allowed diagnosis of CFS with high sensitivity (91%; 95% confidence interval [CI], 57 to 99%) and specificity (71%; 95% CI, 41 to 90%). The positive and negative prognostic values were 71% (95% CI, 41 to 90%) and 91% (95% CI, 57 to 99%), respectively. In the absence of acute infection or chronic inflammation, a high RNase L ratio could distinguish CFS patients from healthy volunteers. Additional large studies and follow-up studies are required to confirm the stability of this high ratio of RNase L isoforms in a CFS group. PMID- 12626459 TI - Elevated levels of matrix metalloproteinase 9 and tissue inhibitor of metalloproteinase 1 during the acute phase of Kawasaki disease. AB - Kawasaki disease (KD) is an acute, self-limiting, multisystem vasculitis of unknown etiology affecting infants and young children. Unless treated promptly with high-dose intravenous gamma globulin and aspirin, patients frequently develop coronary aneurysms. Previously, matrix metalloproteinase 9 (MMP-9), which is secreted complexed to tissue inhibitor of metalloproteinase 1 (TIMP-1), has been implicated in abdominal aortic aneurysm formation. Since the clinical and pathological features of KD include inflammation and weakening of blood vessels, we analyzed acute- and convalescent-phase paired plasma or serum samples from 31 KD patients, 7 patients who did not completely meet the criteria for KD, and 26 non-KD controls (9 febrile and 17 afebrile patients) for pro-MMP-9 (92 kDa) enzyme activity by gelatin zymography and for active MMP-9 (83 kDa), pro-MMP-9, and TIMP-1 protein levels by enzyme-linked immunosorbent assay. Statistical analysis was performed by using Student t tests, linear regression, and the Wilcoxon rank-sum test. Markedly elevated pro-MMP-9 enzymatic activity, pro-MMP-9 protein levels, and TIMP-1 protein levels were found during the acute phase of illness in patients with clinically established KD and in patients who were suspected of having KD but did not meet all of the criteria. There was no significant difference in active MMP-9 levels. Furthermore, pro-MMP-9 and TIMP-1 protein levels were significantly elevated among KD patients, compared to those of febrile and afebrile non-KD controls. The significantly elevated pro-MMP-9 enzyme and protein levels during the acute phase of KD may reflect vascular remodeling or an inflammatory response to a microbial agent, suggesting a pathophysiological role for MMP-9 in coronary aneurysm formation. PMID- 12626463 TI - Nucleoporin p62 antibodies in a case of mixed connective tissue disease. AB - Mixed connective tissue disease is an overlap syndrome characterized by features of different systemic autoimmune diseases and a high titer of U1-snRNP antibodies. We examine here the autoantibodies to nucleoporin p62 in a severe case of mixed connective tissue disease in a young male patient. Thus far, p62 antibodies have mainly been described in cases of primary biliary cirrhosis. We speculate that the presence of p62 antibodies is an indication of a poor prognosis in connective tissue disorders. PMID- 12626462 TI - Assessment of thymic activity in human immunodeficiency virus-negative and positive adolescents by real-time PCR quantitation of T-cell receptor rearrangement excision circles. AB - Circular DNA molecules known as T-cell receptor rearrangement excision circles (TREC) arise during T-cell development and are present in cells that have recently emigrated from the thymus. In cross-sectional studies, the number of peripheral blood lymphocytes bearing TREC decreases with age, consistent with an anatomically demonstrated loss of thymic epithelial tissue. TREC numbers increase following hematopoietic stem cell transplantation and during therapy for human immunodeficiency virus (HIV) infection. Quantitation of TREC has therefore been proposed as a parameter of thymic activity. In this study, we used real-time PCR to quantify TREC in peripheral blood samples obtained longitudinally from HIV seronegative adolescents. TREC values in peripheral blood T cells were very stable throughout adolescence, once thought to be a time of rapid involution of the thymus. In addition, in a cross-sectional analysis, we examined TREC values in a cohort of HIV-positive adolescents and found evidence of ongoing thymopoiesis in perinatally infected individuals, despite lifelong infection. These data demonstrate the utility of TREC assessment in adolescents and that HIV infection does not uniformly result in accelerated thymic involution in childhood. PMID- 12626461 TI - Diagnosis of dengue virus infection by detection of specific immunoglobulin M (IgM) and IgA antibodies in serum and saliva. AB - To evaluate alternative approaches to the serological diagnosis of dengue virus (DEN) infection, the detection of DEN-specific immunoglobulin M (IgM) and IgA antibodies in serum and saliva specimens was assessed in 147 patients with symptoms of DEN infection seen at the Ministry of Health in Nicaragua. Seventy two serum samples were determined to be positive for anti-DEN antibodies by IgM capture enzyme-linked immunosorbent assay, the routine diagnostic procedure. Serum and saliva specimens were obtained from 50 healthy adults as additional controls. IgM was detected in the saliva of 65 of the 72 serum IgM-positive cases, 6 of the 75 serum IgM-negative cases, and none of the control group, resulting in a sensitivity of 90.3% and a specificity of 92.0% and demonstrating that salivary IgM is a useful diagnostic marker for DEN infection. Detection of IgA in serum may be another feasible alternative for the diagnosis of DEN infection, with serum IgA found in 68 (94.4%) of the IgM-positive cases. In contrast, detection of IgA in saliva was not found to be a useful tool for DEN diagnosis in the present study. Further studies of the kinetics of antibody detection in another set of 151 paired acute- and convalescent-phase serum samples showed that DEN-specific IgA antibodies were detected in more acute-phase samples than were IgM antibodies. Thus, we conclude that DEN-specific IgA in serum is a potential diagnostic target. Furthermore, given that saliva is a readily obtainable, noninvasive specimen, detection of DEN-specific salivary IgM should be considered a useful, cheaper diagnostic modality with similar sensitivity and specificity to IgM detection in serum. PMID- 12626464 TI - Interleukin 12 in part regulates gamma interferon release in human whole blood stimulated with Leptospira interrogans. AB - Heat-killed pathogenic Leptospira interrogans serovar rachmati induced the production of gamma interferon (IFN-gamma) and the IFN-gamma-inducing cytokines interleukin-12p40 (IL-12p40) and tumor necrosis factor alpha in human whole blood in vitro. The production of IFN-gamma was largely dependent on IL-12. These data establish that pathogenic leptospires can stimulate the production of type I cytokines involved in cellular immunity. PMID- 12626465 TI - Genetic models in applied physiology. PMID- 12626466 TI - Measuring lung function in mice: the phenotyping uncertainty principle. AB - Measuring lung function in mice is essential for establishing the relevance of murine models to human lung disease. However, making such measurements presents particular technical challenges due to the small size of the animal, particularly with regard to the measurement of respiratory flows. In this review, we examine the various methods currently available for assessment of lung function in mice and contrast them in terms of a concept we call the phenotyping uncertainty principle; each method can be considered to lie somewhere along a continuum on which noninvasiveness must be traded off against experimental control and measurement precision. Unrestrained plethysmography in conscious mice represents the extreme of noninvasiveness and is highly convenient but provides respiratory measures that are so tenuously linked to respiratory mechanics that they cannot be considered as meaningful indicators of lung function. At the other extreme, the measurement of input impedance in anesthetized, paralyzed, tracheostomized mice is precise and specific but requires that an animal be studied under conditions far from natural. In between these two extremes lie methods that sacrifice some precision for a reduction in the level of invasiveness, a promising example being the measurement of transfer impedance in conscious, restrained mice. No method is optimal in all regards; therefore, the appropriate technique to use depends on the application. PMID- 12626467 TI - Effect of water temperature on cooling efficiency during hyperthermia in humans. AB - We evaluated the cooling rate of hyperthermic subjects, as measured by rectal temperature (T(re)), during immersion in a range of water temperatures. On 4 separate days, seven subjects (4 men, 3 women) exercised at 65% maximal oxygen consumption at an ambient temperature of 39 degrees C until T(re) increased to 40 degrees C (45.4 +/- 4.1 min). After exercise, the subjects were immersed in a circulated water bath controlled at 2, 8, 14, or 20 degrees C until T(re) returned to 37.5 degrees C. No difference in cooling rate was observed between the immersions at 8, 14, and 20 degrees C despite the differences in the skin surface-to-water temperature gradient, possibly because of the presence of shivering at 8 and 14 degrees C. Compared with the other conditions, however, the rate of cooling (0.35 +/- 0.14 degrees C/min) was significantly greater during the 2 degrees C water immersion, in which shivering was seldom observed. This rate was almost twice as much as the other conditions (P < 0.05). Our results suggest that 2 degrees C water is the most effective immersion treatment for exercise-induced hyperthermia. PMID- 12626468 TI - Middle cerebral artery blood velocity during intense static exercise is dominated by a Valsalva maneuver. AB - Lifting of a heavy weight may lead to "blackout" and occasionally also to cerebral hemorrhage, indicating pronounced consequences for the blood flow through the brain. We hypothesized that especially strenuous respiratory straining (a Valsalva-like maneuver) associated with intense static exercise would lead to a precipitous rise in mean arterial and central venous pressures and, in turn, influence the middle cerebral artery blood velocity (MCA V(mean)) as a noninvasive indicator of changes in cerebral blood flow. In 10 healthy subjects, MCA V(mean) was evaluated in response to maximal static two-legged exercise performed either with a concomitantly performed Valsalva maneuver or with continued ventilation and also during a Valsalva maneuver without associated exercise (n = 6). During static two-legged exercise, the largest rise for mean arterial pressure and MCA V(mean) was established at the onset of exercise performed with a Valsalva-like maneuver (by 42 +/- 5 mmHg and 31 +/- 3% vs. 22 +/ 6 mmHg and 25 +/- 6% with continued ventilation; P < 0.05). Profound reductions in MCA V(mean) were observed both after exercise with continued ventilation (-29 +/- 4% together with a reduction in the arterial CO(2) tension by -5 +/- 1 Torr) and during the maintained Valsalva maneuver (-21 +/- 3% together with an elevation in central venous pressure to 40 +/- 7 mmHg). Responses to performance of the Valsalva maneuver with and without exercise were similar, reflecting the deterministic importance of the Valsalva maneuver for the central and cerebral hemodynamic response to intense static exercise. Continued ventilation during intense static exercise may limit the initial rise in arterial pressure and may in turn reduce the risk of hemorrhage. On the other hand, blackout during and after intense static exercise may reflect a reduction in cerebral blood flow due to expiratory straining and/or hyperventilation. PMID- 12626469 TI - Intramyocellular triacylglycerol in prolonged cycling with high- and low carbohydrate availability. AB - Vastus lateralis intramyocellular lipid (IMCL) content was assessed by (1)H magnetic resonance spectroscopy before and after prolonged time trial cycling bouts of approximately 3-h duration. Six highly trained male cyclists completed a double-blind, randomized, crossover design of two experimental trials after a strenuous exercise bout and 48 h of high (HC) (9.32 +/- 0.08 g. kg(-1). day(-1)) and low (LC) (0.59 +/- 0.21 g. kg(-1). day(-1)) dietary carbohydrate. Resting IMCL content was significantly higher after LC vs. HC (P < 0.01) and was reduced during exercise by 64 and 57%, respectively. IMCL was not different between conditions after exercise (P > 0.05). The approximately twofold increase in IMCL degradation in LC compared with HC suggests that higher rates of whole body lipid metabolism in LC were in part attributable to a greater IMCL utilization. Four subjects experienced reductions of IMCL in excess of 70% during exercise. To our knowledge, this is the first study to report near depletion of IMCL during prolonged cycling, indicating that IMCL, presumably the triacylglycerol component, may be exhausted by prolonged strenuous exercise. PMID- 12626470 TI - Hypochlorous acid modifies calcium release channel function from skeletal muscle sarcoplasmic reticulum. AB - We have previously demonstrated that H2O2 at millimolar concentrations induces Ca(2+) release from actively loaded sarcoplasmic reticulum (SR) vesicles and induces biphasic [(3)H]ryanodine binding behavior. Considering that hypochlorous acid (HOCl) is a related free radical and has been demonstrated to be a more effective oxidant of proteins, we evaluated the effects of HOCl on sarcoplasmic reticulum Ca(2+)-channel release mechanism. In a concentration-dependent manner, HOCl activates the SR Ca(2+) release channel and induces rapid release of Ca from actively loaded vesicles. HOCl-induced Ca(2+) release is inhibited in the presence of millimolar concentrations of DMSO. High-affinity [(3)H]ryanodine binding is also enhanced at concentrations from 10 to 100 microM. At HOCl concentrations of >100 microM, equilibrium binding is inhibited. HOCl stimulation of binding is inhibited by the addition of dithiothreitol. The direct interaction between HOCl and the Ca(2+) release mechanism was further demonstrated in single channel reconstitution experiments. HOCl, at 20 microM, activated the Ca(2+) release channel after fusion of a SR vesicle to a bilayer lipid membrane. At 40 microM, Ca(2+)-channel activity was inhibited. Pretreatment of SR vesicles with HOCl inhibited the fluorescence development of a fluorogenic probe specific to thiol groups critical to channel function. These results suggest that HOCl at micromolar concentrations can modify SR Ca(2+) handling. PMID- 12626471 TI - Effects of hypoxia on rat airway smooth muscle cell proliferation. AB - Although it is well known that hypoxemia induces pulmonary vasoconstriction and vascular remodeling, due to the proliferation of both vascular smooth muscle cells and fibroblasts, the effects of hypoxemia on airway smooth muscle cells are not well characterized. The present study was designed to assess the in vitro effects of hypoxia (1 or 3% O(2)) on rat airway smooth muscle cell growth and response to mitogens (PDGF and 5-HT). Cell growth was assessed by cell counting and cell cycle analysis. Compared with normoxia (21% O(2)), there was a 42.2% increase in the rate of proliferation of cells exposed to 3% O(2) (72 h, P = 0.006), as well as an enhanced response to PDGF (13.9% increase; P = 0.023) and to 5-HT (17.2% increase; P = 0.039). Exposure to 1% O(2) (72 h) decreased cell proliferation by 21.0% (P = 0.017) and reduced the increase in cell proliferation induced by PGDF and 5-HT by 16.2 and 15.7%, respectively (P = 0.019 and P = 0.011). A significant inhibition in hypoxia-induced cell proliferation was observed after the administration of bisindolylmaleimide GF-109203X (a specific PKC inhibitor) or downregulation of PKC with PMA. Pretreatment with GF-109203X decreased proliferation by 21.5% (P = 0.004) and PMA by 31.5% (P = 0.005). These results show that hypoxia induces airway smooth muscle cell proliferation, which is at least partially dependent on PKC activation. They suggest that hypoxia could contribute to airway remodeling in patients suffering from chronic, severe respiratory diseases. PMID- 12626472 TI - Respiratory effects of the scalene and sternomastoid muscles in humans. AB - Previous studies have shown that in normal humans the change in airway opening pressure (DeltaPao) produced by all the parasternal and external intercostal muscles during a maximal contraction is approximately -18 cmH(2)O. This value is substantially less negative than DeltaPao values recorded during maximal static inspiratory efforts in subjects with complete diaphragmatic paralysis. In the present study, therefore, the respiratory effects of the two prominent inspiratory muscles of the neck, the sternomastoids and the scalenes, were evaluated by application of the Maxwell reciprocity theorem. Seven healthy subjects were placed in a computed tomographic scanner to determine the fractional changes in muscle length during inflation from functional residual capacity to total lung capacity and the masses of the muscles. Inflation induced greater shortening of the scalenes than the sternomastoids in every subject. The inspiratory mechanical advantage of the scalenes thus averaged (mean +/- SE) 3.4 +/- 0.4%/l, whereas that of the sternomastoids was 2.0 +/- 0.3%/l (P < 0.001). However, sternomastoid muscle mass was much larger than scalene muscle mass. As a result, DeltaPao generated by a maximal contraction of either muscle would be 3-4 cmH(2)O, which is about the same as DeltaPao generated by the parasternal intercostals in all interspaces. PMID- 12626473 TI - Perturbations in three medullary nuclei enhance fractionated breathing in awake goats. AB - Our aim was to determine the frequency and characteristics of a fractionated pattern of diaphragm and upper airway muscle activity and airflow during wakefulness and sleep in adult goats. A fractionated breath (FBr) was defined as three or more brief (40-150 ms) interruptions in the diaphragm activity not associated with multiple swallows, eructation, mastication, or movement. During a FBr, the discharge pattern in the diaphragm and upper airway muscles showed complete cycles of inspiration and expiration. Whereas the interval between peak diaphragm activity of the breath preceding the FBr to the first diaphragm peak of the FBr was 15-20% less than the average interval of the preceding five control breaths, the breath-to-breath interval of the five breaths after a FBr did not differ from the control breaths before the FBr event. In normal goats, FBr was evident in only 4 of 18 (22%) awake goats and in only one of these goats during non-rapid eye movement sleep. In 35 goats with implanted microtubules in the medulla, FBr were present in 14 (40%) goats. In these goats with FBr, 78% (11 of 14) had one or more implantations into or near the facial, vestibular, or raphe nuclei. The effect of perturbations in these nuclei is probably nonspecific, because injections into these nuclei with mock cerebrospinal fluid or excitatory amino acid-receptor agonist or antagonist produced both increases and decreases in the frequency of the FBr while not altering their characteristics. Finally, a swallow occurred at the termination or during the first breath after 60% of the FBr. We speculate that the FBr manifest 1) the disruption of a neuronal network, which coordinates breathing and other functions (such as swallowing), utilizing the same anatomic structures, and/or 2) transient changes in synaptic inputs that increase the rate of the normal respiratory rhythm generator or allow an ectopic, anomalous generator to become dominant. PMID- 12626474 TI - Structural and functional differences of the carotid body between DBA/2J and A/J strains of mice. AB - In a previous study, DBA/2J and A/J inbred mice showed extremely different hypoxic ventilatory responses, suggesting variations in their carotid bodies. We have assessed the morphological and functional differences of the carotid bodies in these mice. Histological examination revealed a clearly delineated carotid body only in the DBA/2J mice. Many typical glomus cells and glomeruli appeared in the DBA/2J but not in the A/J mice. The size of the carotid body in the DBA/2J and A/J mice was 6.3 +/- 0.5 x 10(6) and 1.5 +/- 0.3 x 10(6) micro m(3), respectively. The area immunostained for tyrosine hydroxylase, an estimation of the glomus cell quantity, was four times larger in the DBA/2J mice than in the A/J mice. The individual data points in the DBA/2J mice segregated from those in the A/J mice. ACh increased intracellular Ca(2+) in most clusters (81%) of cultured carotid body cells from the DBA/2J mice, but only in 18% of clusters in the A/J mice. These data suggest that genetic determinants account for the strain differences in the structure and function of the carotid body. PMID- 12626475 TI - Bioimpedance analysis: a useful technique for assessing appendicular lean soft tissue mass and distribution. AB - The present study was aimed at evaluating the feasibility and reliability of lower limb skeletal muscle (SM) mass estimates obtained by bioimpedance analysis (BIA). BIA estimates were compared with the estimates obtained by dual-energy X ray absorptiometry (DXA). Ten normal weight and 10 obese women had BIA and DXA evaluations. Lower limb SM mass was then derived from DXA appendicular lean soft tissue estimates. Lower limb SM mass and SM distribution were also estimated from BIA modeling that fits measured resistance values along the leg. SM mass (mean +/ SD) was 5.8 +/- 1.0 kg by BIA vs. 5.8 +/- 1.1 kg by DXA in normal weight subjects and 7.2 +/- 1.4 kg by BIA vs. 7.2 +/- 1.2 kg by DXA in obese subjects. Mean +/- SD of the absolute value of the relative error was 7.0 +/- 3.4 and 5.9 +/- 3.4% in the two groups, respectively. Similar results were obtained by using five resistance values for the analysis. In conclusion, the proposed BIA model provides an adequate means of evaluating appendicular SM mass. PMID- 12626476 TI - Immediate sensory nerve-mediated respiratory responses to irritants in healthy and allergic airway-diseased mice. AB - The immediate responses of the upper respiratory tract (URT) to the irritants acrolein and acetic acid were examined in healthy and allergic airway-diseased C57Bl/6J mice. Acrolein (1.1 ppm) and acetic acid (330 ppm) vapors induced an immediate increase in flow resistance, as measured in the surgically isolated URT of urethane-anesthetized healthy animals. Acrolein, but not acetic acid, induced a small URT vasodilatory response. In awake spontaneously breathing mice, both vapors induced a prolonged pause at the start of expiration (a response mediated via stimulation of nasal trigeminal nerves) and an increase in total respiratory specific airway flow resistance, the magnitude of which was similar to that observed in the isolated URT. Both responses were significantly reduced in animals pretreated with large doses of capsaicin to defunctionalize sensory nerves, strongly suggesting a role for sensory nerves in development of these responses. The breathing pattern and/or obstructive responses were enhanced in mice with ovalbumin-induced allergic airway disease. These results suggest that the primary responses to acrolein and acetic acid vapors are altered breathing patterns and airway obstruction, that sensory nerves play an important role in these responses, and that these responses are enhanced in animals with allergic airway disease. PMID- 12626477 TI - Red blood cell orientation in pulmonary capillaries and its effect on gas diffusion. AB - When alveoli are inflated, the stretched alveolar walls draw their capillaries into oval cross sections. This causes the disk-shaped red blood cells to be oriented near alveolar gas, thereby minimizing diffusion distance. We tested these ideas by measuring red blood cell orientation in histological slides from rapidly frozen rat lungs. High lung inflation did cause the capillaries to have oval cross sections, which constrained the red blood cells within them to flow with their broad sides facing alveolar gas. Low lung inflation stretched alveolar walls less and allowed the capillaries to assume a circular cross section. The circular luminal profile permitted the red blood cells to have their edges facing alveolar gas, which increased the diffusion distance. Using a finite-element method to calculate the diffusing capacity of red blood cells in the broad-side and edge-on orientations, we found that edge-on red blood cells had a 40% lower diffusing capacity. This suggests that, when capillary cross sections become circular, whether through low-alveolar volume or through increased microvascular pressure, the red blood cells are likely to be less favorably oriented for gas exchange. PMID- 12626478 TI - Brain temperature measured by 1H-NMR in conjunction with a lanthanide complex. AB - In vivo data on temperature distributions in the intact brain are scarce, partly due to lack of noninvasive methods for the region of interest. NMR has been exploited for probing a variety of brain activities in vivo noninvasively within the region of interest. Here we report the use of a thulium-based thermometric sensor, infused through the blood, for monitoring absolute temperature in rat brain in vivo by (1)H-NMR and validated by direct temperature measurements with thermocouple wires. Because the (1)H chemical shifts also demonstrate pH sensitivity, detection of multiple resonances was used to measure both temperature and pH simultaneously with high sensitivity. Examination of blood plasma and cerebral spinal fluid samples removed from rats infused with the thermometric sensor suggests that the complex, despite its negative charge, crosses the blood-brain barrier to enter the extracellular milieu. In the future, the thulium-based thermometric sensor may be used for monitoring temperature (and pH) distributions throughout the entire brain, examining response to therapy and evaluating changes induced by alterations in neuronal activity. PMID- 12626480 TI - Invited review: Sleeping flies don't lie: the use of Drosophila melanogaster to study sleep and circadian rhythms. AB - During the past century, flies thoroughly proved their value as an animal model for the study of the genetics of development and basic cell processes. During the past three decades, they have also been extensively used to study the genetics of behavior. For both circadian rhythms and for sleep, flies are helping us to understand the genetic mechanisms that underlie these complex behaviors. Since 1971, discoveries in the fly have led the way to a number of significant discoveries, establishing a mechanistic framework that is now known to be conserved in the mammalian clock. The highlights of this history are described. For sleep, the use of the fly as a model is relatively new, that is, only within the past 2 yr. Nonetheless, studies have already established that two transcription factors alter rest and rest homeostasis. The implications of these advances for the future of sleep research are summarized. PMID- 12626479 TI - Invited review: Identifying new mouse models of cardiovascular disease: a review of high-throughput screens of mutagenized and inbred strains. AB - The mouse is a proven model for studying human disease. Many strains exist that exhibit either natural or engineered genetic variation and thereby enable the elucidation of pathways involved in the development of cardiovascular disease. Although those mouse models have been fundamental to advancing our knowledge base, we are still at an early stage in understanding how genes contribute to complex disorders. There remains a need for new animal models that closely represent human disease. To expedite their development, we have established the Center for New Mouse Models of Heart, Lung, Blood, and Sleep Disorders at The Jackson Laboratory. We are using a phenotype-driven approach to identify mutations leading to atherosclerosis, hypertension, obesity, blood disorders, lung dysfunction, thrombosis, and disordered sleep. Our high-throughput, comprehensive phenotyping draws from two sources for new models: 1) the natural variation among over 40 inbred mouse strains and 2) chemically induced, whole genome mutagenized mice. Here, we review our cardiovascular screens and present some hypertensive, obese, and cardiovascular models identified with this approach. PMID- 12626482 TI - Exercise delays the hypoxic thermal response in rats. AB - Exercise exacerbates acute mountain sickness. In infants and small mammals, hypoxia elicits a decrease in body temperature (Tb) [hypoxic thermal response (HTR)], which may protect against hypoxic tissue damage. We postulated that exercise would counteract the HTR and promote hypoxic tissue damage. Tb was measured by telemetry in rats (n = 28) exercising or sedentary in either normoxia or hypoxia (10% O2, 24 h) at 25 degrees C ambient temperature (Ta). After 24 h of normoxia, rats walked at 10 m/min on a treadmill (30 min exercise, 30 min rest) for 6 h followed by 18 h of rest in either hypoxia or normoxia. Exercising normoxic rats increased Tb ( degrees C) vs. baseline (39.68 +/- 0.99 vs. 38.90 +/ 0.95, mean +/- SD, P < 0.05) and vs. sedentary normoxic rats (38.0 +/- 0.09, P < 0.05). Sedentary hypoxic rats decreased Tb (36.15 +/- 0.97 vs. 38.0 +/- 0.36, P < 0.05) whereas Tb was maintained in the exercising hypoxic rats during the initial 6 h of exercise (37.61 +/- 0.55 vs. 37.72 +/- 1.25, not significant). After exercise, Tb in hypoxic rats reached a nadir similar to that in sedentary hypoxic rats (35.05 +/- 1.69 vs. 35.03 +/- 1.32, respectively). Tb reached its nadir significantly later in exercising hypoxic vs. sedentary hypoxic rats (10.51 +/- 1.61 vs. 5.36 +/- 1.83 h, respectively; P = 0.002). Significantly greater histopathological damage and water contents were observed in brain and lungs in the exercising hypoxic vs. sedentary hypoxic and normoxic rats. Thus exercise early in hypoxia delays but does not prevent the HTR. Counteracting the HTR early in hypoxia by exercise exacerbates brain and lung damage and edema in the absence of ischemia. PMID- 12626484 TI - Age effects on force produced by intrinsic and extrinsic hand muscles and finger interaction during MVC tasks. AB - Finger-pressing forces are produced by activation of the intrinsic hand muscles, which are finger specific, and the extrinsic muscles that connect to multiple fingers. We tested a hypothesis of greater weakening of intrinsic hand muscles with age and quantified associated indexes of finger interaction such as enslaving (force production by unintended fingers) and force deficit (loss of finger force in multifinger tasks compared with single-finger tasks). Twelve young (23-35 yr old) and 12 elderly (70-95 yr old) men and women performed single finger and four-finger maximal pressing tasks, in which force was applied at the proximal phalanges (PP, the intrinsic muscles are major focal force generators) and at the distal phalanges (DP, the extrinsic muscles are focal force generators). The decline in the peak force with age was greater at PP (30%) than at DP (19%). Larger indexes of finger interaction were observed at PP (enslaving = 17.2 +/- 9.4%, force deficit = 36.1 +/- 11.1%) than at DP (enslaving = 14.9 +/- 8.8%, force deficit = 27.7 +/- 10.8%) across ages and genders. We conclude that intrinsic hand muscles show disproportionate weakening with age. The greater indexes of finger interaction in PP tests with greater involvement of intrinsic hand muscles suggest that the finger interactions are predominantly of a central origin across ages and genders. PMID- 12626483 TI - Gender, exercise training, and eNOS expression in porcine skeletal muscle arteries. AB - Our purpose was to determine the effects of gender and exercise training on endothelial nitric oxide synthase (eNOS) and superoxide dismutase (SOD) protein content of porcine skeletal muscle arteries and to evaluate the role of 17beta estradiol (E2) in these effects. We measured eNOS and SOD content with immunoblots and immunohistochemistry in femoral and brachial arteries of trained and sedentary male and female pigs and measured estrogen receptor (ER) mRNA and alpha-ER and beta-ER protein in aortas of male and female pigs. Results indicate that female arteries contain more eNOS than male arteries and that exercise training increases eNOS content independent of gender. Male and female pigs expressed similar levels of alpha-ER mRNA and protein and similar amounts beta-ER protein in their arteries. E2 concentrations as measured by RIA were 180 +/- 34 pg/ml in male sera and approximately 5 pg/ml in female sera, and neither was changed by training. However, bioassay indicated that biologically active estrogen equivalent to only 35 +/- 5 pg/ml was present in male sera. E2 in female pigs, whether measured by RIA or bioassay, was approximately 24 pg/ml at peak estrous and 2 pg/ml on day 5 diestrus. The free fraction of E2 in sera did not explain the low measurements, relative to RIA, of E2. We conclude that 1). gender has significant influence on eNOS and SOD content of porcine skeletal muscle arteries; 2). the effects of gender and exercise training vary among arteries of different anatomic origin; 3). male sera contains compounds that cause RIA to overestimate circulating estrogenic activity; and 4). relative to human men, the male pig is not biologically estrogenized by high levels of E2 reported by RIA, whereas in female pigs E2 levels are lower than in the blood of human women. PMID- 12626485 TI - Coronary smooth muscle reactivity to muscarinic stimulation after ischemia reperfusion in porcine myocardial infarction. AB - This study tested whether ischemia-reperfusion alters coronary smooth muscle reactivity to vasoconstrictor stimuli such as those elicited by an adventitial stimulation with methacholine. In vitro studies were performed to assess the reactivity of endothelium-denuded infarct-related coronary arteries to methacholine (n = 18). In addition, the vasoconstrictor effects of adventitial application of methacholine to left anterior descending (LAD) coronary artery was assessed in vivo in pigs submitted to 2 h of LAD occlusion followed by reperfusion (n = 12), LAD deendothelization (n = 11), or a sham operation (n = 6). Endothelial-dependent vasodilator capacity of infarct-related LAD was assessed by intracoronary injection of bradykinin (n = 13). In vitro, smooth muscle reactivity to methacholine was unaffected by ischemia-reperfusion. In vivo, baseline methacholine administration induced a transient and reversible drop in coronary blood flow (9.6 +/- 4.6 to 1.9 +/- 2.6 ml/min, P < 0.01), accompanied by severe left ventricular dysfunction. After ischemia-reperfusion, methacholine induced a prolonged and severe coronary blood flow drop (9.7 +/- 7.0 to 3.4 +/- 3.9 ml/min), with a significant delay in recovery (P < 0.001). Endothelial denudation mimics in part the effects of methacholine after ischemia reperfusion, and intracoronary bradykinin confirmed the existence of endothelial dysfunction. Infarct-related epicardial coronary artery shows a delayed recovery after vasoconstrictor stimuli, because of appropriate smooth muscle reactivity and impairment of endothelial-dependent vasodilator capacity. PMID- 12626486 TI - Active upper airway closure during induced central apneas in lambs is complete at the laryngeal level only. AB - We tested the hypotheses that active upper airway closure during induced central apneas in nonsedated lambs 1). is complete and occurs at the laryngeal level and 2). is not due to stimulation of the superior laryngeal nerves (SLN). Five newborn lambs were surgically instrumented to record thyroarytenoid (TA) muscle (glottal constrictor) electromyographic (EMG) activity with supra- and subglottal pressures. Hypocapnic and nonhypocapnic central apneas were induced before and after SLN sectioning in the five lambs. A total of 174 apneas were induced, 116 before and 58 after sectioning of the internal branch of the SLN (iSLN). Continuous TA EMG activity was observed in 88% of apneas before iSLN section and in 87% of apneas after iSLN section. A transglottal pressure different from zero was observed in all apneas with TA EMG activity, with a mean subglottal pressure of 4.3 +/- 0.8 cmH2O before and 4.7 +/- 0.7 cmH2O after iSLN section. Supraglottal pressure was consistently atmospheric. Sectioning of both iSLNs had no effects on the results. We conclude that upper airway closure during induced central apneas in lambs is active, complete, and occurs at the glottal level only. Consequently, a positive subglottal pressure is maintained throughout the apnea. Finally, this complete active glottal closure is independent from laryngeal afferent innervation. PMID- 12626487 TI - Evaluation of musculotendinous stiffness in prepubertal children and adults, taking into account muscle activity. AB - Musculotendinous (MT) stiffness of the triceps surae (TS) muscle group was quantified in 28 prepubertal children (7-10 yr) by using quick-release movements at different levels of submaximal contractions. Surface electromyograms (EMG) of each part of the TS and of the tibialis anterior were also recorded. A stiffness index, defined as the slope of the angular stiffness-torque relationship (SIMT Torque), was used to quantify changes in MT stiffness with age. Results showed a significant decrease in SIMT-Torque with age, ranging from 4.02 +/- 0.29 to 2.88 +/- 0.31 rad-1 for the youngest to the oldest children. Because an increase in stiffness with age was expected due to the maturation of elastic tissues, overactivation of the TS was suspected to contribute to the higher SIMT-Torque values found in the youngest children. TS EMG-torque analyses confirmed that neuromuscular efficiency was significantly lower for the 7- or 8-yr-old children compared with 10-yr-old children, notably due to a higher degree of tibialis anterior coactivation found in the youngest children. Thus the stiffness index originally defined as the slope of the angular stiffness-EMG relationship increased significantly with age toward adult values. The results underlined the necessity to take into account the capacities of muscle activation to quantify changes in elastic properties of muscles, when those capacities are suspected to be altered. PMID- 12626488 TI - Increased susceptibility to Pseudomonas aeruginosa infection under hindlimb unloading conditions. AB - It has been reported that spaceflight conditions alter the immune system and resistance to infection [Belay T, Aviles H, Vance M, Fountain K, and Sonnenfeld G. J Allergy Clin Immunol 170: 262-268, 2002; Hankins WR and Ziegelschmid JF. In: Biomedical Results of Apollo. Washington, DC: NASA, 1975, p. 43-81. (NASA Spec. Rep. SP-368)]. Ground-based models, including the hindlimb-unloading model, have become important tools for increasing understanding of how spaceflight conditions can influence physiology. The objective of the present study was to determine the effect of hindlimb unloading on the susceptibility of mice to Pseudomonas aeruginosa infection. Hindlimb-unloaded and control mice were subcutaneously infected with 1 LD50 of P. aeruginosa. Survival, bacterial organ load, and antibody and corticosterone levels were compared among the groups. Hindlimb unloading had detrimental effects for infected mice. Animals in the hindlimb unloaded group, compared with controls, 1). showed significantly increased mortality and reduced time to death, 2). had increased levels of corticosterone, and 3). were much less able to clear bacteria from the organs. These results suggest that hindlimb unloading may induce the production of corticosterone, which may play a critical role in the modulation of the immune system leading to increased susceptibility to P. aeruginosa infection. PMID- 12626490 TI - CO2 does not affect passive exercise ventilatory decline. AB - Breathing increases abruptly at the start of passive exercise, stimulated by afferent feedback from the moving limbs, and declines toward a steady-state hyperpnea as exercise continues. This decline has been attributed to decreased arterial CO2 levels and adaptation in afferent feedback; however, the relative importance of these two mechanisms is unknown. To address this issue, we compared ventilatory responses to 5 min of passive leg extension exercise performed on 10 awake human subjects (6 men and 4 women) in isocapnic and poikilocapnic conditions. End-tidal Pco2 decreased significantly during poikilocapnic (Delta = 1.5 +/- 0.5 Torr, P < 0.001), but not isocapnic, passive exercise. Despite this difference, the ventilatory responses to passive exercise were not different between the two conditions. Using the fast changes in ventilation at the start (5.46 +/- 0.40 l/min, P < 0.001) and end (3.72 +/- 0.33 l/min, P < 0.001) of passive exercise as measures of the drive to breathe from afferent feedback, we found a decline of 68%. We conclude that the decline in ventilation during passive exercise is due to an adaptation in the afferent feedback from the moving limbs, not a decline in CO2 levels. PMID- 12626489 TI - EPR spectroscopic detection of free radical outflow from an isolated muscle bed in exercising humans. AB - There is no direct evidence to support the contention that contracting skeletal muscle and/or associated vasculature generates free radicals in exercising humans. The unique combination of isolated quadriceps exercise and the measurement of femoral arterial and venous free radical concentrations with the use of electron paramagnetic resonance (EPR) spectroscopy enabled this assumption to be tested in seven healthy men. Application of ex vivo spin trapping using alpha-phenyl-tert-butylnitrone (PBN) resulted in the detection of oxygen- or carbon-centered free radicals (a(N) = 1.38 +/- 0.01 mT and a(beta)(H) = 0.17 +/ 0.01 mT, where a(N) and a(beta)(H) are the nitrogen and beta-hydrogen coupling constants, respectively) with consistently higher EPR signal intensities of the PBN spin adduct observed in the venous compared with the arterial circulation (P < 0.05). Incremental exercise further increased the venoarterial intensity difference [85 +/- 58 arbitrary units (AU) at 24 +/- 6% maximal work rate (WR(max)) vs. 387 +/- 214 AU at 69 +/- 7% WR(max); P < 0.05]. When combined with measured changes in femoral venous blood flow (Q), this resulted in a net adduct outflow of 130 +/- 118 and 1,146 +/- 582 AU/min (P < 0.05), which was positively associated with leg oxygen uptake (r(2) = 0.47, P < 0.05) and Q (r(2) = 0.47, P < 0.05). These results provide the first evidence for oxygen- or carbon-centered free radical outflow from an active muscle bed in humans. PMID- 12626491 TI - Adaptation to chronic length change in explanted airway smooth muscle. AB - It has been shown that airway smooth muscle in vitro is able to maintain active force over a large length range by adaptation in the absence of periodic stimulations at 4 degrees C (Wang L, Pare PD, and Seow CY. J Appl Physiol 90: 734 740, 2001). In this study, we show that such adaptation also takes place at body temperature and that long-term adaptation results in irreversible functional change in the muscle that could lead to airway hyperresponsiveness. Rabbit tracheal muscle explants were passively maintained at shortened and in situ length for 3 and 7-8 days in culture media; the length-tension relationship was then examined. The length associated with maximal force generation decreased by 10.5 +/- 3.8% (SE) after 3 days and 37.7 +/- 8.5% after 7 or 8 days of passive shortening. At day 3, the left shift in the length-tension curve due to adaptation at short lengths was reversible by readapting the muscle at a longer length. The shift was, however, not completely reversible after 7 days. The results suggest that long-term adaptation of airway smooth muscle could lead to increased muscle stiffness and force-generating ability at short lengths. Under in vivo condition, this could translate into resistance to stretch-induced relaxation and excessive airway narrowing. PMID- 12626492 TI - Effect of surface tension of mucosal lining liquid on upper airway mechanics in anesthetized humans. AB - Upper airway (UA) patency may be influenced by surface tension (gamma) operating within the (UAL). We examined the role of gamma of UAL in the maintenance of UA patency in eight isoflurane-anesthetized supine human subjects breathing via a nasal mask connected to a pneumotachograph attached to a pressure delivery system. We evaluated 1). mask pressure at which the UA closed (Pcrit), 2). UA resistance upstream from the site of UA collapse (RUS), and 3). mask pressure at which the UA reopened (Po). A multiple pressure-transducer catheter was used to identify the site of airway closure (velopharyngeal in all subjects). UAL samples (0.2 microl) were collected, and the gamma of UAL was determined by using the "pull-off force" technique. Studies were performed before and after the intrapharyngeal instillation of 5 ml of exogenous surfactant (Exosurf, Glaxo Smith Kline). The gamma of UAL decreased from 61.9 +/- 4.1 (control) to 50.3 +/- 5.0 mN/m (surfactant; P < 0.02). Changes in Po, RUS, and Po - Pcrit (change = control - surfactant) were positively correlated with changes in gamma (r2 > 0.6; P < 0.02) but not with changes in Pcrit (r2 = 0.4; P > 0.9). In addition, mean peak inspiratory airflow (no flow limitation) significantly increased (P < 0.04) from 0.31 +/- 0.06 (control) to 0.36 +/- 0.06 l/s (surfactant). These findings suggest that gamma of UAL exerts a force on the UA wall that hinders airway opening. Instillation of exogenous surfactant into the UA lowers the gamma of UAL, thus increasing UA patency and augmenting reopening of the collapsed airway. PMID- 12626493 TI - Galphaq-coupled receptor internalization specifically induced by Galphaq signaling. Regulation by EBP50. AB - In the present report, we investigated the effect of ezrin-radixin-moesin-binding phosphoprotein 50 (EBP50) expression on the agonist-induced internalization of the thromboxane A(2) beta receptor (TPbeta receptor). Interestingly, we found that EBP50 almost completely blocked TPbeta receptor internalization, which could not be reversed by overexpression of G protein-coupled receptor (GPCR) kinases and arrestins. Because we recently demonstrated that EBP50 can bind to and inhibit Galpha(q), we next studied whether Galpha(q) signaling could induce TPbeta receptor internalization, addressing the long standing question about the relationship between GPCR signaling and their internalization. Expression of a constitutively active Galpha(q) mutant (Galpha(q)-R183C) resulted in a robust internalization of the TPbeta receptor, which was unaffected by expression of dominant negative mutants of arrestin-2 and -3, but inhibited by expression of EBP50 or dynamin-K44A, a dominant negative mutant of dynamin. Phospholipase Cbeta and protein kinase C did not appear to significantly contribute to internalization of the TPbeta receptor, suggesting that Galpha(q) induces receptor internalization through a phospholipase Cbeta- and protein kinase C independent pathway. Surprisingly, there appears to be specificity in Galpha protein-mediated GPCR internalization. Galpha(q)-R183C also induced the internalization of CXCR4 (Galpha(q)-coupled), whereas it failed to do so for the beta(2)-adrenergic receptor (Galpha(s)-coupled). Moreover, Galpha(s)-R201C, a constitutively active form of Galpha(s), had no effect on internalization of the TPbeta, CXCR4, and beta(2)-adrenergic receptors. Thus, we showed that Galpha protein signaling can lead to internalization of GPCRs, with specificity in both the Galpha proteins and GPCRs that are involved. Furthermore, a new function has been described for EBP50 in its capacity to inhibit receptor endocytosis. PMID- 12626494 TI - Strategy to discriminate between high and low affinity bindings of human immunodeficiency virus, type 1 integrase to viral DNA. AB - The last decade has contributed to our understanding of the three-dimensional structure of the human immunodeficiency virus, type 1 (HIV-1) integrase (IN) and to the description of how the enzyme catalyzes the viral DNA integration into the host DNA. Recognition of the viral DNA termini by IN is sequence-specific, and that of the host DNA does not require particular sequence, although in physicochemical studies IN fails to discriminate between the two interactions. Here, such discrimination was allowed thanks to a model system using designed oligonucleotides and peptides as binding structures. Spectroscopic (circular dichroism, NMR, and fluorescence anisotropy) techniques and biochemical (enzymatic and filter binding) assays clearly indicated that the amphipathic helix alpha4, located at the catalytic domain surface, is responsible for the specific high affinity binding of the enzyme to viral DNA. Analogues of the alpha4 peptide having increased helicity and still bearing the biologically relevant lysines 156 and 159 on the DNA binding face, and oligonucleotides conserving an intact attachment site, are required to achieve high affinity complexes (Kd of 1.5 nm). Data corroborate previous in vivo results obtained with mutated viruses. PMID- 12626495 TI - Macroautophagy is required for multicellular development of the social amoeba Dictyostelium discoideum. AB - Macroautophagy is a mechanism employed by eukaryotic cells to recycle non essential cellular components during starvation, differentiation, and development. Two conjugation reactions related to ubiquitination are essential for autophagy: Apg12p conjugation to Apg5p, and Apg8p conjugation to the lipid phosphatidylethanolamine. These reactions require the action of the E1-like enzyme, Apg7p, and the E2-like enzymes, Apg3p and Apg10p. In Dictyostelium, development is induced by starvation, conditions under which autophagy is required for survival in yeast and plants. We have identified Dictyostelium homologues of 10 budding yeast autophagy genes. We have generated mutations in apg5 and apg7 that produce defects typically associated with an abrogation of autophagy. Mutants are not grossly affected in growth, but survival during nitrogen starvation is severely reduced. Starved mutant cells show little turnover of cellular constituents by electron microscopy, whereas wild-type cells show significant cytoplasmic degradation and reduced organelle number. Bulk protein degradation during starvation-induced development is reduced in the autophagy mutants. Development is aberrant; the autophagy mutants do not aggregate in plaques on bacterial lawns, but they do proceed further in development on nitrocellulose filters, forming defective fruiting bodies. The autophagy mutations are cell autonomous, because wild-type cells in a chimaera do not rescue development of the autophagy mutants. We have complemented the mutant phenotypes by expression of the cognate gene fused to green fluorescent protein. A green fluorescent protein fusion of the autophagosome marker Apg8 mislocalizes in the two autophagy mutants. We show that the Apg5-Apg12 conjugation system is conserved in Dictyostelium. PMID- 12626496 TI - Conserved Asp684 in transmembrane segment M6 of the plant plasma membrane P-type proton pump AHA2 is a molecular determinant of proton translocation. AB - The mechanism of proton pumping by P-type H(+)-ATPases is still unclear. In the plant P-type plasma membrane H(+)-ATPase AHA2, two charged residues, Arg(655) and Asp(684), are conserved in transmembrane segments M5 and M6, respectively, a region that has been shown be contribute to ion coordination in related P-type ATPases. Substitution of Arg(655) with either alanine or aspartate resulted in mutant enzymes exhibiting a significant shift in the P-type ATPase E(1)P-E(2)P conformational equilibrium. The mutant proteins accumulated in the E(1)P conformation, but were capable of conducting proton transport. This points to an important role of Arg(655) in the E(1)P-E(2)P conformational transition. The presence of a carboxylate moiety at position Asp(684) proved essential for coupling between initial proton binding and proton pumping. The finding that the carboxylate side chain of Asp(684) contributes to the proton-binding site and appears to function as an absolutely essential proton acceptor along the proton transport pathway is discussed in the context of a possible proton pumping mechanism of P-type H(+)-ATPases. PMID- 12626497 TI - Structure-function relations of interactions between Na,K-ATPase, the gamma subunit, and corticosteroid hormone-induced factor. AB - Corticosteroid hormone-induced factor (CHIF) and the gamma subunit of the Na,K ATPase (gamma) are two members of the FXYD family whose function has been elucidated recently. CHIF and gamma interact with the Na+ pump and alter its kinetic properties, in different ways, which appear to serve their specific physiological roles. Although functional interactions with the Na,K-ATPase have been clearly demonstrated, it is not known which domains and which residues interact with the alpha and/or beta subunits and affect the pump kinetics. The current study provides the first systematic analysis of structure-function relations of CHIF and gamma. It is demonstrated that the stability of detergent solubilized complexes of CHIF and gamma with alpha and/or beta subunits is determined by the trans-membrane segments, especially three residues that may be involved in hydrophobic interactions. The transmembrane segments also determine the opposite effects of CHIF and gamma on the Na+ affinity of the pump, but the amino acids involved in this functional effect are different from those responsible for stable interactions with alpha. PMID- 12626498 TI - Biochemical properties of CikA, an unusual phytochrome-like histidine protein kinase that resets the circadian clock in Synechococcus elongatus PCC 7942. AB - We recently described the cikA (circadian input kinase A) gene, whose product supplies environmental information to the circadian oscillator in the cyanobacterium Synechococcus elongatus PCC 7942. CikA possesses three distinct domains: a GAF, a histidine protein kinase (HPK), and a receiver domain similar to those of the response regulator family. To determine how CikA functions in providing circadian input, we constructed modified alleles to tag and truncate the protein, allowing analysis of each domain individually. CikA covalently bound bilin chromophores in vitro, even though it lacks the expected ligand residues, and the GAF domain influenced but did not entirely account for this function. Full-length CikA and truncated variants that carry the HPK domain showed autophosphorylation activity. Deletion of the GAF domain or the N-terminal region adjacent to GAF dramatically reduced autophosphorylation, whereas elimination of the receiver domain increased activity 10-fold. Assays to test phosphorelay from the HPK to the cryptic receiver domain, which lacks the conserved aspartyl residue that serves as a phosphoryl acceptor in response regulators, were negative. We propose that the cryptic receiver is a regulatory domain that interacts with an unknown protein partner to modulate the autokinase activity of CikA but does not work as bona fide receiver domain in a phosphorelay. PMID- 12626499 TI - Specific amino acid substitutions determine the differential contribution of the N- and C-terminal domains of insulin-like growth factor (IGF)-binding protein-5 in binding IGF-I. AB - We have previously reported that two highly conserved amino acids in the C terminal domain of rat insulin-like growth factor-binding protein (IGFBP)-5, Gly(203) and Gln(209), are involved in binding to insulin-like growth factor (IGF)-1. Here we report that mutagenesis of both amino acids simultaneously (C Term mutant) results in a cumulative effect and an even greater reduction in IGF I binding: 30-fold measured by solution phase IGF binding assay and 10-fold by biosensor analysis. We compared these reductions in ligand binding to the effects of specific mutations of five amino acids in the N-terminal domain (N-Term mutant), which had previously been shown by others to cause a very large reduction in IGF-I binding (). Our results confirm this as the major IGF-binding site. To prove that the mutations in either N- or C-Term were specific for IGF-I binding, we carried out CD spectroscopy and showed that these alterations did not lead to gross conformational changes in protein structure for either mutant. Combining these mutations in both domains (N+C-Term mutant) has a cumulative effect and leads to a 126-fold reduction in IGF-I binding as measured by biosensor. Furthermore, the equivalent mutations in the C terminus of rat IGFBP-2 (C-Term 2) also results in a significant reduction in IGF-I binding, suggesting that the highly conserved Gly and Gln residues have a conserved IGF-I binding function in all six IGFBPs. Finally, although these residues lie within a major heparin-binding site in IGFBP-5 and -3, we also show that the mutations in C-Term have no effect on heparin binding. PMID- 12626500 TI - Transforming growth factor-beta 1 potentiates amyloid-beta generation in astrocytes and in transgenic mice. AB - Accumulation of the amyloid-beta peptide (Abeta) in the brain is crucial for development of Alzheimer's disease. Expression of transforming growth factor beta1 (TGF-beta1), an immunosuppressive cytokine, has been correlated in vivo with Abeta accumulation in transgenic mice and recently with Abeta clearance by activated microglia. Here, we demonstrate that TGF-beta1 drives the production of Abeta40/42 by astrocytes leading to Abeta production in TGF-beta1 transgenic mice. First, TGF-beta1 induces the overexpression of the amyloid precursor protein (APP) in astrocytes but not in neurons, involving a highly conserved TGF beta1-responsive element in the 5'-untranslated region (+54/+74) of the APP promoter. Second, we demonstrated an increased release of soluble APP-beta which led to TGF-beta1-induced Abeta generation in both murine and human astrocytes. These results demonstrate that TGF-beta1 potentiates Abeta production in human astrocytes and may enhance the formation of plaques burden in the brain of Alzheimer's disease patients. PMID- 12626502 TI - Liver cell polyploidization: a pivotal role for binuclear hepatocytes. AB - Polyploidy is a general physiological process indicative of terminal differentiation. During liver growth, this process generates the appearance of tetraploid (4n) and octoploid (8n) hepatocytes with one or two nuclei. The onset of polyploidy in the liver has been recognized for quite some time; however, the cellular mechanisms that govern it remain unknown. In this report, we observed the sequential appearance during liver growth of binuclear diploid (2 x 2n) and mononuclear 4n hepatocytes from a diploid hepatocyte population. To identify the cell cycle modifications involved in hepatocyte polyploidization, mitosis was then monitored in primary cultures of rat hepatocytes. Twenty percent of mononuclear 2n hepatocytes failed to undergo cytokinesis with no observable contractile movement of the ring. This process led to the formation of binuclear 2 x 2n hepatocytes. This tetraploid condition following cleavage failure did not activate the p53-dependent checkpoint in G1. In fact, binuclear hepatocytes were able to proceed through S phase, and the formation of a bipolar spindle during mitosis constituted the key step leading to the genesis of two mononuclear 4n hepatocytes. Finally, we studied the duplication and clustering of centrosomes in the binuclear hepatocyte. These cells exhibited two centrosomes in G1 that were duplicated during S phase and then clustered by pairs at opposite poles of the cell during metaphase. This event led only to mononuclear 4n progeny and maintained the tetraploidy status of hepatocytes. PMID- 12626501 TI - The molecular neighborhood of subunit 8 of yeast mitochondrial F1F0-ATP synthase probed by cysteine scanning mutagenesis and chemical modification. AB - The detailed membrane topography and neighboring polypeptides of subunit 8 in yeast mitochondrial ATP synthase have been determined using a combination of cysteine scanning mutagenesis and chemical modification. 46 single cysteine substitution mutants encompassing the length of the subunit 8 protein were constructed by site-directed mutagenesis. Expression of each cysteine variant in yeast lacking endogenous subunit 8 restored respiratory phenotype to cells and had little measurable effect on ATP hydrolase function. The exposure of each introduced cysteine residue to the aqueous environment was assessed in isolated mitochondria using the fluorescent thiol-modifying probe fluorescein 5-maleimide. The first 14 and last 13 amino acids of subunit 8 were accessible to fluorescein 5-maleimide in osmotically lysed mitochondria and are thus extrinsic to the lipid bilayer, indicating a 21-amino acid transmembrane span. The C-terminal region of subunit 8 was partially occluded by other ATP synthase subunits, especially in a small region surrounding Val-40 that was demonstrated to play an important role in maintaining the stability of the F(1)-F(0) interaction. Cross-linking using heterobifunctional reagents revealed the proximity of subunit 8 to subunits b, d, and f in the matrix and to subunits b, f, and 6 in the intermembrane space. A disulfide bridge was also formed between subunit 8(F7C) or (M10C) and residue Cys 23 of subunit 6, demonstrating a close interaction between these two hydrophobic membrane subunits and confirming the location of the N termini of each in the intermembrane space. We conclude that subunit 8 is an integral component of the stator stalk of yeast mitochondrial F(1)F(0)-ATP synthase. PMID- 12626504 TI - Intra-mitochondrial poly(ADP-ribosylation) contributes to NAD+ depletion and cell death induced by oxidative stress. AB - Poly(ADP-ribosylation), primarily via poly(ADP-ribose) polymerase-1 (PARP-1), is a pluripotent cellular process important for maintenance of genomic integrity and RNA transcription in cells. However, during conditions of oxidative stress and energy depletion, poly(ADP-ribosylation) paradoxically contributes to mitochondrial failure and cell death. Although it has been presumed that poly(ADP ribosylation) within the nucleus mediates this pathologic process, PARP-1 and other poly(ADP-ribosyltransferases) are also localized within mitochondria. To this end, the presence of PARP-1 and poly(ADP-ribosylation) were verified within mitochondrial fractions from primary cortical neurons and fibroblasts. Inhibition of poly(ADP-ribosylation) within the mitochondrial compartment preserved transmembrane potential (DeltaPsi(m)), NAD(+) content, and cellular respiration, prevented release of apoptosis-inducing factor, and reduced neuronal cell death triggered by oxidative stress. Treatment with liposomal NAD(+) also preserved DeltaPsi(m) and cellular respiration during oxidative stress. Furthermore, inhibition of poly(ADP-ribosylation) prevented intranuclear localization of apoptosis-inducing factor and protected neurons from excitotoxic injury; and PARP 1 null fibroblasts were protected from oxidative stress-induced cell death. Collectively these data suggest that poly(ADP-ribosylation) compartmentalized to the mitochondria can be converted from a homeostatic process to a mechanism of cell death when oxidative stress is accompanied by energy depletion. These data implicate intra-mitochondrial poly(ADP-ribosylation) as an important therapeutic target for central nervous system and other diseases associated with oxidative stress and energy failure. PMID- 12626505 TI - Control of electron transport in photosystem I by the iron-sulfur cluster FX in response to intra- and intersubunit interactions. AB - Photosystem I (PS I) is a transmembranal multisubunit complex that mediates light induced electron transfer from plactocyanine to ferredoxin. The electron transfer proceeds from an excited chlorophyll a dimer (P700) through a chlorophyll a (A0), a phylloquinone (A1), and a [4Fe-4S] iron-sulfur cluster FX, all located on the core subunits PsaA and PsaB, to iron-sulfur clusters FA and FB, located on subunit PsaC. Earlier, it was attempted to determine the function of FX in the absence of FA/B mainly by chemical dissociation of subunit PsaC. However, not all PsaC subunits could be removed from the PS I preparations by this procedure without partially damaging FX. We therefore removed subunit PsaC by interruption of the psaC2 gene of PS I in the cyanobacterium Synechocystis sp. PCC 6803. Cells could not grow under photosynthetic conditions when subunit PsaC was deleted, yet the PsaC-deficient mutant cells grew under heterotrophic conditions and assembled the core subunits of PS I in which light-induced electron transfer from P700 to A1 occurred. The photoreduction of FX was largely inhibited, as seen from direct measurement of the extent of electron transfer from A1 to FX. From the crystal structure it can be seen that the removal of subunits PsaC, PsaD, and PsaE in the PsaC-deficient mutant resulted in the braking of salt bridges between these subunits and PsaB and PsaA and the formation of a net of two negative surface charges on PsaA/B. The potential induced on FX by these surface charges is proposed to inhibit electron transport from the quinone. In the complete PS I complex, replacement of a cysteine ligand of FX by serine in site-directed mutation C565S/D566E in subunit PsaB caused an approximately 10-fold slow down of electron transfer from the quinone to FX without much affecting the extent of this electron transfer compared with wild type. Based on these and other results, we propose that FX might have a major role in controlling electron transfer through PS I. PMID- 12626503 TI - The Shank family of postsynaptic density proteins interacts with and promotes synaptic accumulation of the beta PIX guanine nucleotide exchange factor for Rac1 and Cdc42. AB - The Shank/ProSAP family of multidomain proteins is known to play an important role in organizing synaptic multiprotein complexes. Here we report a novel interaction between Shank and beta PIX, a guanine nucleotide exchange factor for the Rac1 and Cdc42 small GTPases. This interaction is mediated by the PDZ domain of Shank and the C-terminal leucine zipper domain and the PDZ domain-binding motif at the extreme C terminus of beta PIX. Shank colocalizes with beta PIX at excitatory synaptic sites in cultured neurons. In brain, Shank forms a complex with beta PIX and beta PIX-associated signaling molecules including p21 associated kinase (PAK), an effector kinase of Rac1/Cdc42. Importantly, overexpression of Shank in cultured neurons promotes synaptic accumulation of beta PIX and PAK. Considering the involvement of Rac1 and PAK in spine dynamics, these results suggest that Shank recruits beta PIX and PAK to spines for the regulation of postsynaptic structure. PMID- 12626506 TI - The hexokinase of the hyperthermophile Thermoproteus tenax. ATP-dependent hexokinases and ADP-dependent glucokinases, teo alternatives for glucose phosphorylation in Archaea. AB - The phosphorylation of glucose by different sugar kinases plays an essential role in Archaea because of the absence of a phosphoenolpyruvate-dependent transferase system characteristic for Bacteria. In the genome of the hyperthermophilic Archaeon Thermoproteus tenax a gene was identified with sequence similarity to glucokinases of the so-called ROK family (repressor protein, open reading frame, sugar kinase). The T. tenax enzyme, like the recently described ATP-dependent "glucokinase" from Aeropyrum pernix, shows the typical broad substrate specificity of hexokinases catalyzing not only phosphorylation of glucose but also of other hexoses such as fructose, mannose, or 2-deoxyglucose, and thus both enzymes represent true hexokinases. The T. tenax hexokinase shows strikingly low if at all any regulatory properties and thus fulfills no important control function at the beginning of the variant of the Embden-Meyerhof-Parnas pathway in T. tenax. Transcript analyses reveal that the hxk gene of T. tenax is cotranscribed with an upstream located orfX, which codes for an 11-kDa protein of unknown function. Growth-dependent studies and promoter analyses suggest that post-transcriptional RNA processing might be involved in the generation of the monocistronic hxk message, which is observed only under heterotrophic growth conditions. Data base searches revealed T. tenax hexokinase homologs in some archaeal, few eukaryal, and many bacterial genomes. Phylogenetic analyses confirm that the archaeal hexokinase is a member of the so-called ROK family, which, however, should be referred to as ROK group because it represents a group within the bacterial glucokinase fructokinase subfamily II of the hexokinase family. Thus, archaeal hexokinases represent a second major group of glucose phosphorylating enzymes in Archaea beside the recently described archaeal ADP dependent glucokinases, which were recognized as members of the ribokinase family. The distribution of the two types of sugar kinases, differing in their cosubstrate as well as substrate specificity, within Archaea is discussed on the basis of physiological constraints of the respective organisms. PMID- 12626507 TI - Calcium/calmodulin modulation of olfactory and rod cyclic nucleotide-gated ion channels. AB - Cyclic nucleotide-gated (CNG) ion channels mediate sensory transduction in olfactory sensory neurons and retinal photoreceptor cells. In these systems, internal calcium/calmodulin (Ca2+/CaM) inhibits CNG channels, thereby having a putative role in sensory adaptation. Functional differences in Ca2+/CaM-dependent inhibition depend on the different subunit composition of olfactory and rod CNG channels. Recent evidence shows that three subunit types (CNGA2, CNGA4, and CNGB1b) make up native olfactory CNG channels and account for the fast inhibition of native channels by Ca2+/CaM. In contrast, two subunit types (CNGA1 and CNGB1) appear sufficient to mirror the native properties of rod CNG channels, including the inhibition by Ca2+/CaM. Within CNG channel tetramers, specific subunit interactions also mediate Ca2+/CaM-dependent inhibition. In olfactory CNGA2 channels, Ca2+/CaM binds to an N-terminal region and disrupts an interaction between the N- and C-terminal regions, causing inhibition. Ca2+/CaM also binds the N-terminal region of CNGB1 subunits and disrupts an intersubunit, N- and C terminal interaction between CNGB1 and CNGA1 subunits in rod channels. However, the precise N- and C-terminal regions that form these interactions in olfactory channels are different from those in rod channels. Here, we will review recent advances in understanding the subunit composition and the mechanisms and roles for Ca2+/CaM-dependent inhibition in olfactory and rod CNG channels. PMID- 12626508 TI - Hypertonicity activates Na+/H+ exchange through Janus kinase 2 and calmodulin. AB - The type 1 sodium-hydrogen exchanger (NHE-1) is a ubiquitous electroneutral membrane transporter that is activated by hypertonicity in many cells. NHE-1 may be an important pathway for Na(+) entry during volume restoration, yet the molecular mechanisms underlying the osmotic regulation of NHE-1 are poorly understood. In the present study we conducted a screen for important signaling molecules that could be involved in hypertonicity-induced activation of NHE-1 in CHO-K1 cells. Hypertonicity rapidly activated NHE-1 in a concentration-dependent manner as assessed by proton microphysiometry and by measurements of intracellular pH on a FLIPR (fluorometric imaging plate reader). Inhibitors of Ca(2+)/calmodulin (CaM) and Janus kinase 2 (Jak2) attenuated this activation, whereas neither calcium chelation nor inhibitors of protein kinase C, the Ras ERK1/2 pathway, Src kinase, and Ca(2+)/calmodulin-dependent enzymes had significant effects. Hypertonicity also resulted in the rapid tyrosine phosphorylation of Jak2 and STAT3 (the major substrate of Jak2) and CaM. Phosphorylation of Jak2 and CaM were blocked by AG490, an inhibitor of Jak2. Immunoprecipitation studies showed that hypertonicity stimulates the assembly of a signaling complex that includes CaM, Jak2, and NHE-1. Formation of the complex could be blocked by AG490. Thus, we propose that hypertonicity induces activation of NHE-1 in CHO-K1 cells in large part through the following pathway: hypertonicity --> Jak2 phosphorylation and activation --> tyrosine phosphorylation of CaM --> association of CaM with NHE-1 --> NHE-1 activation. PMID- 12626509 TI - Filament formation of MSF-A, a mammalian septin, in human mammary epithelial cells depends on interactions with microtubules. AB - Septins are a family of conserved proteins implicated in a variety of cellular functions such as cytokinesis and vesicle trafficking, but their properties and modes of action are largely unknown. Here we now report findings of immunocytochemical and biochemical characterization of a mammalian septin, MSF-A. Using an antibody specific for MSF subfamily proteins, MSF-A was found to be expressed predominantly in mammary human mammary epithelial cells (HMEC). MSF-A was associated with microtubules in interphase HMEC cells as it localized with the mitotic spindle and the bundle of microtubule at midzone during mitosis. Biochemical analysis revealed direct binding of MSF-A with polymerized tubulin through its central region containing guanine nucleotide-interactive motifs. GTPase activity, however, was not required for the association. Conditions that disrupt the microtubule network also disrupted the MSF-A-containing filament structure, resulting in a punctate cytoplasmic pattern. Depletion of MSF-A using small interfering RNAs caused incomplete cell division and resulted in the accumulation of binucleated cells. Unlike Nedd5, an MSF mutant deficient in GTPase activity forms filament indistinguishable from that of the wild type in COS cells. These results strongly suggest that septin filaments may interact not only with actin filaments but also with microtubule networks and that GTPase activity of MSF-A is not indispensable to incorporation of MSF-A into septin filaments. PMID- 12626510 TI - Sas4 and Sas5 are required for the histone acetyltransferase activity of Sas2 in the SAS complex. AB - The SAS2 gene is involved in transcriptional silencing in Saccharomyces cerevisiae. Based on its primary sequence, the Sas2 protein is predicted to be a member of the MYST family of histone acetyltransferases (HATs). Sas2 forms a complex with Sas4 and Sas5, which are required for its silencing function. Here we show that recombinant Sas2 has HAT activity that absolutely requires Sas4 and is stimulated by Sas5. The recombinant SAS complex acetylates H4 lysine 16 and H3 lysine 14. Furthermore, a purified SAS complex from yeast shows similar activity and specificity. In contrast to other MYST HATs, neither the recombinant nor the native SAS complex acetylated nucleosomal histones under conditions that were optimum for acetylating free histones. Finally, although the SAS subunits interact genetically and physically with Asf1, a histone deposition factor, association of H3 and H4 with Asf1 blocks their acetylation by the SAS complex, raising the possibility that the SAS HAT complex may acetylate free histones prior to their deposition onto DNA by Asf1 or CAF-I. PMID- 12626512 TI - Mechanisms of VE-cadherin processing and degradation in microvascular endothelial cells. AB - VE-cadherin is an endothelial-specific cadherin that plays important roles in vascular morphogenesis and growth control. To investigate the mechanisms by which endothelial cells regulate cadherin cell surface levels, a VE-cadherin mutant containing the non-adhesive interleukin-2 (IL-2) receptor extracellular domain and the VE-cadherin cytoplasmic tail (IL-2R-VE-cadcyto) was expressed in microvascular endothelial cells. Expression of the IL-2R-VE-cadcyto mutant resulted in the internalization of endogenous VE-cadherin and in a dramatic decrease in endogenous VE-cadherin levels. The internalized VE-cadherin co localized with early endosomes, and the lysosomal inhibitor chloroquine dramatically inhibited the down-regulation of VE-cadherin in cells expressing the IL-2R-VE-cadcyto mutant. Chloroquine treatment also resulted in the accumulation of a VE-cadherin fragment lacking the beta-catenin binding domain of the VE cadherin cytoplasmic tail. The formation of the VE-cadherin fragment could be prevented by treating endothelial cells with proteasome inhibitors. Furthermore, inhibition of the proteasome prevented VE-cadherin internalization and inhibited the disruption of endothelial intercellular junctions by the IL-2RVE-cadcyto mutant. These results provide new insights into the mechanisms of VE-cadherin processing and degradation in microvascular endothelial cells. PMID- 12626511 TI - Analysis of myosin heavy chain functionality in the heart. AB - Comparison of mammalian cardiac alpha- and beta-myosin heavy chain isoforms reveals 93% identity. To date, genetic methodologies have effected only minor switches in the mammalian cardiac myosin isoforms. Using cardiac-specific transgenesis, we have now obtained major myosin isoform shifts and/or replacements. Clusters of non-identical amino acids are found in functionally important regions, i.e. the surface loops 1 and 2, suggesting that these structures may regulate isoform-specific characteristics. Loop 1 alters filament sliding velocity, whereas Loop 2 modulates actin-activated ATPase rate in Dictyostelium myosin, but this remains untested in mammalian cardiac myosins. Alpha --> beta isoform switches were engineered into mouse hearts via transgenesis. To assess the structural basis of isoform diversity, chimeric myosins in which the sequences of either Loop 1+Loop 2 or Loop 2 of alpha-myosin were exchanged for those of beta-myosin were expressed in vivo. 2-fold differences in filament sliding velocity and ATPase activity were found between the two isoforms. Filament sliding velocity of the Loop 1+Loop 2 chimera and the ATPase activities of both loop chimeras were not significantly different compared with alpha-myosin. In mouse cardiac isoforms, myosin functionality does not depend on Loop 1 or Loop 2 sequences and must lie partially in other non homologous residues. PMID- 12626514 TI - Domains in tropoelastin that mediate elastin deposition in vitro and in vivo. AB - Elastic fiber assembly is a complicated process involving multiple different proteins and enzyme activities. However, the specific protein-protein interactions that facilitate elastin polymerization have not been defined. To identify domains in the tropoelastin molecule important for the assembly process, we utilized an in vitro assembly model to map sequences within tropoelastin that facilitate its association with fibrillin-containing microfibrils in the extracellular matrix. Our results show that an essential assembly domain is located in the C-terminal region of the molecule, encoded by exons 29-36. Fine mapping studies using an exon deletion strategy and synthetic peptides identified the hydrophobic sequence in exon 30 as a major functional element in this region and suggested that the assembly process is driven by the propensity of this sequence to form beta-sheet structure. Tropoelastin molecules lacking the C terminal assembly domain expressed as transgenes in mice did not assemble nor did they interfere with assembly of full-length normal mouse elastin. In addition to providing important information about elastin assembly in general, the results of this study suggest how removal or alteration of the C terminus through stop or frameshift mutations might contribute to the elastin-related diseases supravalvular aortic stenosis and cutis laxa. PMID- 12626515 TI - Protein-tyrosine phosphatase alpha, RPTP alpha, is a Helicobacter pylori VacA receptor. AB - Helicobacter pylori vacuolating cytotoxin, VacA, induces vacuolation, mitochondrial damage, cytochrome c release, and apoptosis of gastric epithelial cells. To detect gastric proteins that serve as VacA receptors, we used VacA co immunoprecipitation techniques following biotinylation of the cell surface and identified p250, a receptor-like protein-tyrosine phosphatase beta (RPTP beta) as a VacA-binding protein (Yahiro, K., Niidome, T., Kimura, M., Hatakeyama, T., Aoyagi, H., Kurazono, H., Imagawa, K., Wada, A., Moss, J., and Hirayama, T. (1999) J. Biol. Chem. 274, 36693-36699). VacA causes vacuolation of G401 cells, a human kidney tumor cell line, although they do not express RPTP beta. By co immunoprecipitation with VacA, we identified p140 as a potential receptor in those cells. p140 purified by chromatography on a peanut agglutinin affinity matrix contained internal amino acid sequences of RGEENTDYVNASFIDGYRQK and AEGILDVFQTVK, which are identical to those in RPTP alpha. The peptide mass fingerprinting of p140 by time of flight-MS analysis also supported this identification. Treatment of G401 cells with RPTP alpha-morpholino antisense oligonucleotide before exposure to toxin inhibited vacuolation. These data suggest that RPTP alpha acts as a receptor for VacA in G401 cells. Thus, two receptor tyrosine phosphatases, RPTP alpha and RPTP beta, serve as VacA receptors. PMID- 12626516 TI - Stepping and stretching. How kinesin uses internal strain to walk processively. AB - The ability of kinesin to travel long distances on its microtubule track without dissociating has led to a variety of models to explain how this remarkable degree of processivity is maintained. All of these require that the two motor domains remain enzymatically "out of phase," a behavior that would ensure that, at any given time, one motor is strongly attached to the microtubule. The maintenance of this coordination over many mechanochemical cycles has never been explained, because key steps in the cycle could not be directly observed. We have addressed this issue by applying several novel spectroscopic approaches to monitor motor dissociation, phosphate release, and nucleotide binding during processive movement by a dimeric kinesin construct. Our data argue that the major effect of the internal strain generated when both motor domains of kinesin bind the microtubule is to block ATP from binding to the leading motor. This effect guarantees the two motor domains remain out of phase for many mechanochemical cycles and provides an efficient and adaptable mechanism for the maintenance of processive movement. PMID- 12626517 TI - The binding sites on human heme oxygenase-1 for cytochrome p450 reductase and biliverdin reductase. AB - Human heme oxygenase-1 (hHO-1) catalyzes the NADPH-cytochrome P450 reductase dependent oxidation of heme to biliverdin, CO, and free iron. The biliverdin is subsequently reduced to bilirubin by biliverdin reductase. Earlier kinetic studies suggested that biliverdin reductase facilitates the release of biliverdin from hHO-1 (Liu, Y., and Ortiz de Montellano, P. R. (2000) J. Biol. Chem. 275, 5297-5307). We have investigated the binding of P450 reductase and biliverdin reductase to truncated, soluble hHO-1 by fluorescence resonance energy transfer and site-specific mutagenesis. P450 reductase and biliverdin reductase bind to truncated hHO-1 with Kd = 0.4 +/- 0.1 and 0.2 +/- 0.1 microm, respectively. FRET experiments indicate that biliverdin reductase and P450 reductase compete for binding to truncated hHO-1. Mutation of surface ionic residues shows that hHO-1 residues Lys18, Lys22, Lys179, Arg183, Arg198, Glu19, Glu127, and Glu190 contribute to the binding of cytochrome P450 reductase. The mutagenesis results and a computational analysis of the protein surfaces partially define the binding site for P450 reductase. An overlapping binding site including Lys18, Lys22, Lys179, Arg183, and Arg185 is similarly defined for biliverdin reductase. These results confirm the binding of biliverdin reductase to hHO-1 and define binding sites of the two reductases. PMID- 12626518 TI - The yersinia virulence factor YopM forms a novel protein complex with two cellular kinases. AB - Pathogenic Yersinia contain a virulence plasmid that encodes genes for intracellular effectors, which neutralize the host immune response. One effector, YopM, is necessary for Yersinia virulence, but its function in host cells is unknown. To identify potential cellular pathways affected by YopM, proteins that co-immunoprecipitate with YopM in mammalian cells were isolated and identified by mass spectrometry. Results demonstrate that two kinases, protein kinase C-like 2 (PRK2) and ribosomal S6 protein kinase 1 (RSK1), interact directly with YopM. These two kinases associate only when YopM is present, and expression of YopM in cells stimulates the activity of both kinases. RSK1 is activated directly by interaction with YopM, and RSK1 kinase activity is required for YopM-stimulated PRK2 activity. YopM activation of RSK1 occurs independently of the actions of YopJ on the MAPK pathway. YopM is also required for Yersinia-induced changes in RSK1 mobility in infected macrophage cells. These results identify the first intracellular targets of YopM and suggest YopM acts to stimulate the activity of PRK2 and RSK1. PMID- 12626519 TI - Direct interaction of Ca2+/calmodulin inhibits histone deacetylase 5 repressor core binding to myocyte enhancer factor 2. AB - Myocyte enhancer factor 2 (MEF2) proteins play a pivotal role in the differentiation of cardiac and skeletal muscle cells. MEF2 factors are regulated by histone deacetylase enzymes such as histone deacetylase 5 (HDAC5). HDAC5 in turn is responsive to Ca(2+) signaling mediated by the intracellular calcium sensor calmodulin. Here a combination of proteolytic fragmentation, matrix assisted laser desorption ionization mass spectrometry, Edman degradation, circular dichroism, gel filtration, and surface plasmon resonance studies is utilized to define and characterize a stable core domain of HDAC5 and to examine its interactions with MEF2a and calmodulin. Results from real time binding experiments provide evidence for direct interaction of Ca(2+)/calmodulin with HDAC5 inhibiting MEF2a association with this enzyme. PMID- 12626520 TI - Taurolithocholic acid exerts cholestatic effects via phosphatidylinositol 3 kinase-dependent mechanisms in perfused rat livers and rat hepatocyte couplets. AB - Taurolithocholic acid (TLCA) is a potent cholestatic agent. Our recent work suggested that TLCA impairs hepatobiliary exocytosis, insertion of transport proteins into apical hepatocyte membranes, and bile flow by protein kinase Cepsilon (PKCepsilon)-dependent mechanisms. Products of phosphatidylinositol 3 kinases (PI3K) stimulate PKCepsilon. We studied the role of PI3K for TLCA-induced cholestasis in isolated perfused rat liver (IPRL) and isolated rat hepatocyte couplets (IRHC). In IPRL, TLCA (10 micromol/liter) impaired bile flow by 51%, biliary secretion of horseradish peroxidase, a marker of vesicular exocytosis, by 46%, and the Mrp2 substrate, 2,4-dinitrophenyl-S-glutathione, by 95% and stimulated PI3K-dependent protein kinase B, a marker of PI3K activity, by 154% and PKCepsilon membrane binding by 23%. In IRHC, TLCA (2.5 micromol/liter) impaired canalicular secretion of the fluorescent bile acid, cholylglycylamido fluorescein, by 50%. The selective PI3K inhibitor, wortmannin (100 nmol/liter), and the anticholestatic bile acid tauroursodeoxycholic acid (TUDCA, 25 micromol/liter) independently and additively reversed the effects of TLCA on bile flow, exocytosis, organic anion secretion, PI3K-dependent protein kinase B activity, and PKCepsilon membrane binding in IPRL. Wortmannin also reversed impaired bile acid secretion in IRHC. These data strongly suggest that TLCA exerts cholestatic effects by PI3K- and PKCepsilon-dependent mechanisms that are reversed by tauroursodeoxycholic acid in a PI3K-independent way. PMID- 12626521 TI - Serine 338 phosphorylation is dispensable for activation of c-Raf1. AB - Numerous extracellular agonists induce consecutive stimulation of Ras guanine nucleotide exchange factors, Ras and c-Raf1, as the starting point of the intracellular mitogen-activated protein kinase cascade. Recent data point to a more complex reaction pattern of this simple sequence. This study was aimed at elucidating the activation process of endogenous c-Raf1 in U937 cells. Treatment of permeabilized U937 cells with the nonhydrolyzable nucleotide guanosine 5'-3-O (thio)triphosphate (GTPgammaS) induced prolonged stimulation of Ras and c-Raf1 activity. Intriguingly, both signaling proteins expressed differential responses toward specific inhibitors of phosphoinositide 3-kinases and tyrosine kinases, which indicates diverse signaling reactions feeding into Ras and cRaf-1. Phosphorylation of c-Raf1 serine 338 by p21-activated kinase has been recently reported to contribute to phosphoinositide 3-kinase-dependent activation of c Raf1. However, in U937 cells stimulation of c-Raf1 activity by GTPgammaS did not correlate with p21-activated kinase activity and Ser-338 phosphorylation. Thus Ser-338 phosphorylation appears dispensable for c-Raf1 activation under the conditions used. Together these data deny an essential role for serine 338 phosphorylation in c-Raf1 activation and disclose divergent signaling connections of Ras and c-Raf1 in U937 cells. PMID- 12626522 TI - Structure-to-function relationship of mini-lipoxygenase, a 60-kDa fragment of soybean lipoxygenase-1 with lower stability but higher enzymatic activity. AB - Lipoxygenase-1 (Lox-1) is a member of the lipoxygenase family, a class of dioxygenases that take part in the metabolism of polyunsatured fatty acids in eukaryotes. Tryptic digestion of soybean Lox-1 is known to produce a 60 kDa fragment, termed "mini-Lox," which shows enhanced catalytic efficiency and higher membrane-binding ability than the native enzyme (Maccarrone, M., Salucci, M. L., van Zadelhoff, G., Malatesta, F., Veldink, G. Vliegenthart, J. F. G., and Finazzi Agro, A. (2001) Biochemistry 40, 6819-6827). In this study, we have investigated the stability of mini-Lox in guanidinium hydrochloride and under high pressure by fluorescence and circular dichroism spectroscopy. Only a partial unfolding could be obtained at high pressure in the range 1-3000 bar at variance with guanidinium hydrochloride. However, in both cases a reversible denaturation was observed. The denaturation experiments demonstrate that mini-Lox is a rather unstable molecule, which undergoes a two-step unfolding transition at moderately low guanidinium hydrochloride concentration (0-4.5 m). Both chemical- and physical-induced denaturation suggest that mini-Lox is more hydrated than Lox-1, an observation also confirmed by 1-anilino-8-naphthalenesulfonate (ANS) binding studies. We have also investigated the occurrence of substrate-induced changes in the protein tertiary structure by dynamic fluorescence techniques. In particular, eicosatetraynoic acid, an irreversible inhibitor of lipoxygenase, has been used to mimic the effect of substrate binding. We demonstrated that mini-Lox is indeed characterized by much larger conformational changes than those occurring in the native Lox-1 upon binding of eicosatetraynoic acid. Finally, by both activity and fluorescence measurements we have found that 1-anilino-8-naphthalenesulfonate has access to the active site of mini-Lox but not to that of intact Lox-1. These findings strongly support the hypothesis that the larger hydration of mini-Lox renders this molecule more flexible, and therefore less stable. PMID- 12626523 TI - Potential role of microsomal prostaglandin E synthase-1 in tumorigenesis. AB - Microsomal prostaglandin E2 synthase-1 (mPGES-1) is a stimulus-inducible enzyme that functions downstream of cyclooxygenase (COX)-2 in the PGE2-biosynthetic pathway. Given the accumulating evidence that COX-2-derived PGE2 participates in the development of various tumors, including colorectal cancer, we herein examined the potential involvement of mPGES-1 in tumorigenesis. Immunohistochemical analyses demonstrated the expression of both COX-2 and mPGES 1 in human colon cancer tissues. HCA-7, a human colorectal adenocarcinoma cell line that displays COX-2- and PGE2-dependent proliferation, expressed both COX-2 and mPGES-1 constitutively. Treatment of HCA-7 cells with an mPGES-1 inhibitor or antisense oligonucleotide attenuated, whereas overexpression of mPGES-1 accelerated, PGE2 production and cell proliferation. Moreover, cotransfection of COX-2 and mPGES-1 into HEK293 cells resulted in cellular transformation manifested by colony formation in soft agar culture and tumor formation when implanted subcutaneously into nude mice. cDNA array analyses revealed that this mPGES-1-directed cellular transformation was accompanied by changes in the expression of a variety of genes related to proliferation, morphology, adhesion, and the cell cycle. These results collectively suggest that aberrant expression of mPGES-1 in combination with COX-2 can contribute to tumorigenesis. PMID- 12626525 TI - Benefit versus risk. PMID- 12626524 TI - Inhibition of epithelial ductal branching in the prostate by sonic hedgehog is indirectly mediated by stromal cells. AB - Sonic hedgehog (Shh), a vertebrate homologue of the Drosophila segment-polarity gene hedgehog, has been reported to play an important role during normal development of various tissues. Abnormal activities of Shh signaling pathway have been implicated in tumorigenesis such as basal cell carcinomas and medulloblastomas. Here we show that Shh signaling negatively regulates prostatic epithelial ductal morphogenesis. In organotypic cultures of developing rat prostates, Shh inhibited cell proliferation and promoted differentiation of luminal epithelial cells. The expression pattern of Shh and its receptors suggests a paracrine mechanism of action. The Shh receptors Ptc1 (Patched1) and Ptc2 were found to be expressed in prostatic stromal cells adjacent to the epithelium, where Shh itself was produced. This paracrine model was confirmed by co-culturing the developing prostate in the presence of stromal cells transfected with a vector expressing a constitutively active form of Smoothened, the real effector of the Shh signaling pathway. Furthermore, expression of activin A and TGF-beta1 that were shown previously to inhibit prostatic epithelial branching was up-regulated following Shh treatment in the organotypic cultures. Taken together, these results suggest that Shh negatively regulates prostatic ductal branching indirectly by acting on the surrounding stromal cells, at least partly via up-regulating expression of activin A and TGF-beta1. PMID- 12626527 TI - Cutting edge: impairment of dendritic cells and adaptive immunity by Ebola and Lassa viruses. AB - Acute infection of humans with Ebola and Lassa viruses, two principal etiologic agents of hemorrhagic fevers, often results in a paradoxical pattern of immune responses: early infection, characterized by an outpouring of inflammatory mediators such as TNF-alpha, IL-1 beta, and IL-6, vs late stage infections, which are associated with poor immune responses. The mechanisms underlying these diverse outcomes are poorly understood. In particular, the role played by cells of the innate immune system, such as dendritic cells (DC), is not known. In this study, we show that Ebola and Lassa viruses infect human monocyte-derived DC and impair their function. Monocyte-derived DC exposed to either virus fail to secrete proinflammatory cytokines, do not up-regulate costimulatory molecules, and are poor stimulators of T cells. These data represent the first evidence for a mechanism by which Ebola and Lassa viruses target DC to impair adaptive immunity. PMID- 12626528 TI - Cutting edge: Toll-like receptor 9 expression is not required for CpG DNA-aided cross-presentation of DNA-conjugated antigens but essential for cross-priming of CD8 T cells. AB - Covalent linkage of immunostimulatory CpG DNA to OVA results in CpG DNA-aided cross-presentation of OVA by dendritic cells (DCs). In vivo, cross-presentation is conditional for cross-priming of OVA-specific CD8 T cells. In this study, we investigated the involvement of the CpG DNA receptor Toll-like receptor (TLR)9 in CpG DNA-aided cross-presentation and cross-priming. Although CpG DNA-aided cross presentation is not altered in TLR9-deficient cells, TLR9 is required for maturation of APC allowing cross-priming, as resulting in CTL function. These findings imply that TLR9 does not trigger endocytosis of CpG-OVA conjugates, but activates DCs downstream of endocytosis. PMID- 12626529 TI - Cutting edge: CD40 engagement eliminates the need for Bruton's tyrosine kinase in B cell receptor signaling for NF-kappa B. AB - The Tec kinase Bruton's tyrosine kinase (Btk) represents a key intermediary for B cell receptor (BCR) signaling. Btk mutation produces B cell deficiency in mice with X-linked immunodeficiency (xid), and surface Ig-mediated responses of mature B cells are seriously deranged. The central role that Btk plays in directing downstream events produced by BCR engagement is demonstrated by the complete failure of NF-kappa B induction and cellular proliferation following anti-Ig treatment of B cells obtained from xid mice. In this study, we report that the block in BCR signaling produced by Btk mutation is reversed by CD40 engagement. Prior treatment with CD40 ligand normalized subsequent responses of xid B cells to BCR cross-linking, so that typical outcomes of BCR signaling such as NF-kappa B activation and cell cycle progression occurred in a Btk-independent fashion. These results demonstrate that a specific genetic lesion interrupting BCR mediated intracellular signaling is circumvented through stimulation of CD40. PMID- 12626530 TI - Cutting edge: Mycobacterium tuberculosis blocks Ca2+ signaling and phagosome maturation in human macrophages via specific inhibition of sphingosine kinase. AB - One-third of the world's population is infected with Mycobacterium tuberculosis (Mtb), and three million people die of tuberculosis each year. Following its ingestion by macrophages (MPs), Mtb inhibits the maturation of its phagosome, preventing progression to a bactericidal phagolysosome. Phagocytosis of Mtb is uncoupled from the elevation in MP cytosolic Ca(2+) that normally accompanies microbial ingestion, resulting in inhibition of phagosome-lysosome fusion and increased intracellular viability. This study demonstrates that the mechanism responsible for this failure of Ca(2+)-dependent phagosome maturation involves mycobacterial inhibition of MP sphingosine kinase. Thus, inhibition of sphingosine kinase directly contributes to survival of Mtb within human MPs and represents a novel molecular mechanism of pathogenesis. PMID- 12626531 TI - Endogenous dendritic cells are required for amplification of T cell responses induced by dendritic cell vaccines in vivo. AB - Dendritic cells (DCs) loaded in vitro with Ag are used as cellular vaccines to induce Ag-specific immunity. These cells are thought to be responsible for direct stimulation of Ag-specific T cells, which may subsequently mediate immunity. In this study, in transgenic mouse models with targeted MHC class II expression specifically on DCs, we show that the DC vaccine is responsible only for partial CD4(+) T cell activation, but to obtain optimal expansion of T cells in vivo, participation of endogenous (resident) DCs, but not endogenous B cells, is crucial. Transfer of Ag to endogenous DCs seems not to be mediated by simple peptide diffusion, but rather by DC-DC interaction in lymph nodes as demonstrated by histological analysis. In contrast, injection of apoptotic or necrotic DC vaccines does not induce T cell responses, but rather represents an immunological null event, which argues that viability of DC vaccines can be crucial for initial triggering of T cells. We propose that viable DCs from the DC vaccine must migrate to the draining lymph nodes and initiate a T cell response, which thereafter requires endogenous DCs that present transferred Ag in order induce optimal T cell expansion. These results are of specific importance with regard to the applicability of DC vaccinations in tumor patients, where the function of endogenous DCs is suppressed by either tumors or chemotherapy. PMID- 12626532 TI - Unresponsiveness to lymphoid-mediated signals at the neonatal follicular dendritic cell precursor level contributes to delayed germinal center induction and limitations of neonatal antibody responses to T-dependent antigens. AB - The factors limiting neonatal and infant IgG Ab responses to T-dependent Ags are only partly known. In this study, we assess how these B cell responses are influenced by the postnatal development of the spleen and lymph node microarchitecture. When BALB/c mice were immunized with alum-adsorbed tetanus toxoid at various stages of their immune development, a major functional maturation step for induction of serum IgG, Ab-secreting cells, and germinal center (GC) responses was identified between the second and the third week of life. This correlated with the development of the follicular dendritic cell (FDC) network, as mature FDC clusters only appeared at 2 wk of age. Adoptive transfer of neonatal splenocytes into adult SCID mice rapidly induced B cell follicles and FDC precursor differentiation into mature FDC, indicating effective recruitment and signaling capacity of neonatal B cells. In contrast, adoptive transfer of adult splenocytes into neonatal SCID mice induced primary B cell follicles without any differentiation of mature FDC and failed to correct limitations of tetanus toxoid-induced GC. Thus, unresponsiveness to lymphoid-mediated signals at the level of neonatal FDC precursors delays FDC maturation and GC induction, thus limiting primary Ab-secreting cell responses to T-dependent Ags in early postnatal life. PMID- 12626533 TI - IL-1 receptor-associated kinase 1 regulates susceptibility to organ-specific autoimmunity. AB - Infections often precede the development of autoimmunity. Correlation between infection with a specific pathogen and a particular autoimmune disease ranges from moderately strong to quite weak. This lack of correspondence suggests that autoimmunity may result from microbial activation of a generic, as opposed to pathogen-specific host-defense response. The Toll-like receptors, essential to host recognition of microbial invasion, signal through a common, highly conserved pathway, activate innate immunity, and control adaptive immune responses. To determine the influence of Toll/IL-1 signaling on the development of autoimmunity, the responses of wild-type (WT) mice and IL-1R-associated kinase 1 (IRAK1)-deficient mice to induction of experimental autoimmune encephalomyelitis were compared. C57BL/6 and B6.IRAK1-deficient mice were immunized with MOG 35 55/CFA or MOG 35-55/CpG DNA/IFA. WT animals developed severe disease, whereas IRAK1-deficient mice were resistant to experimental autoimmune encephalomyelitis, exhibiting little or no CNS inflammation. IRAK1-deficient T cells also displayed impaired Th1 development, particularly during disease induction, despite normal TCR signaling. These results suggest that IRAK1 and the Toll/IL-1 pathway play an essential role in T cell priming, and demonstrate one means through which innate immunity can control subsequent development of autoimmunity. These findings may also help explain the association between antecedent infection and the development or exacerbations of some autoimmune diseases. PMID- 12626534 TI - Conditional expression of murine Flt3 ligand leads to expansion of multiple dendritic cell subsets in peripheral blood and tissues of transgenic mice. AB - The analysis of the development and function of distinct subsets of murine dendritic cells (DC) has been hampered by the limited number of these cells in vivo. To circumvent this limitation we have developed a conditional transgenic mouse model for producing large numbers of DC. We used the tetracycline-inducible system to conditionally express murine Flt3 ligand (FL), a potent hemopoietic growth factor that promotes the differentiation and mobilization of DC. Acute treatment (96 h) of the transgenic animals with the tetracycline analog doxycycline (DOX) promoted an approximately 200-fold increase in serum levels of FL without affecting the number of circulating DC. However, within 1 wk of DOX treatment, the relative number of DC in peripheral blood increased from approximately 8 to approximately 40%. Interestingly, both the levels of FL and the number of DC remained elevated for at least 9 mo with continual DOX treatment. Chronic treatment of the mice with DOX led to dramatic increases in the number of DC in multiple tissues without any apparent pathological consequences. Most DC populations were expanded, including immature and mature DC, myeloid (CD11c(+)CD11b(+)CD8a(-)), lymphoid (CD11c(+)CD11b(-)CD8a(+)), and the recently defined plasmacytoid (pDC) subsets. Finally, transplantation of BM from green fluorescent protein-expressing mice into lethally irradiated transgenic mice followed by subsequent DOX treatment led to expansion of green fluorescent protein-labeled DC. The transgenic mice described here should thus provide a readily available source of multiple DC subsets and should facilitate the analysis of their role in homeostasis and disease. PMID- 12626535 TI - Low activation threshold as a mechanism for ligand-independent signaling in pre-T cells. AB - Pre-TCR complexes are thought to signal in a ligand-independent manner because they are constitutively targeted to lipid rafts. We report that ligand independent signaling is not a unique capability of the pre-TCR complex. Indeed, the TCR alpha subunit restores development of pT alpha-deficient thymocytes to the CD4(+)CD8(+) stage even in the absence of conventional MHC class I and class II ligands. Moreover, we found that pre-TCR and alpha beta TCR complexes exhibit no appreciable difference in their association with lipid rafts, suggesting that ligand-independence is a function of the CD4(-)CD8(-) (DN) thymocytes in which pre-TCR signaling occurs. In agreement, we found that only CD44(-)CD25(+) DN thymocytes (DN3) enabled activation of extracellular signal-regulated kinases by the pre-TCR complex. DN thymocytes also exhibited a lower signaling threshold relative to CD4(+)CD8(+) thymocytes, which was associated with both the markedly elevated lipid raft content of their plasma membranes and more robust capacitative Ca(2+) entry. Taken together these data suggest that cell autonomous, ligand-independent signaling is primarily a property of the thymocytes in which pre-TCR signaling occurs. PMID- 12626536 TI - The size of the synaptic cleft and distinct distributions of filamentous actin, ezrin, CD43, and CD45 at activating and inhibitory human NK cell immune synapses. AB - In this study, we report the organization of cytoskeletal and large transmembrane proteins at the inhibitory and activating NK cell immunological or immune synapse (IS). Filamentous actin accumulates at the activating, but not the inhibitory, NK cell IS. However, surprisingly, ezrin and the associated protein CD43 are excluded from the inhibitory, but not the activating, NK cell IS. This distribution of ezrin and CD43 at the inhibitory NK cell IS is similar to that previously seen at the activating T cell IS. CD45 is also excluded from the inhibitory, but not activating, NK cell IS. In addition, electron microscopy reveals wide and narrow domains across the synaptic cleft. Target cell HLA-C, located by immunogold labeling, clusters where the synaptic cleft spans the size of HLA-C bound to the inhibitory killer Ig-like receptor. These data are consistent with assembly of the NK cell IS involving a combination of cytoskeletal-driven mechanisms and thermodynamics favoring the organization of receptor/ligand pairs according to the size of their extracellular domains. PMID- 12626537 TI - Forced expression of the Fc receptor gamma-chain renders human T cells hyperresponsive to TCR/CD3 stimulation. AB - High level expression of Fc epsilon RI gamma chain replaces the deficient TCR zeta-chain and contributes to altered TCR/CD3-mediated signaling abnormalities in T cells of patients with systemic lupus erythematosus. Increased responsiveness to Ag has been considered to lead to autoimmunity. To test this concept, we studied early signaling events and IL-2 production in fresh cells transfected with a eukaryotic expression vector encoding the Fc epsilon RI gamma gene. We found that the overexpressed Fc epsilon RI gamma chain colocalizes with the CD3 epsilon chain on the surface membrane of T cells and that cross-linking of the new TCR/CD3 complex leads to a dramatic increase of intracytoplasmic calcium concentration, protein tyrosine phosphorylation, and IL-2 production. We observed that overexpression of Fc epsilon RI gamma is associated with increased phosphorylation of Syk kinase, while the endogenous TCR zeta-chain is down regulated. We propose that altered composition of the CD3 complex leads to increased T cell responsiveness to TCR/CD3 stimulation and sets the biochemical grounds for the development of autoimmunity. PMID- 12626538 TI - T cell activation in vivo targets diacylglycerol kinase alpha to the membrane: a novel mechanism for Ras attenuation. AB - Diacylglycerol kinase (DGK) phosphorylates diacylglycerol to produce phosphatidic acid, leading to decreased and increased levels, respectively, of these two lipid messengers that play a central role in T cell activation. Nine DGK isoforms, grouped into five subtypes, are found in higher organisms; all contain a conserved C-terminal domain and at least two cysteine-rich motifs of unknown function. In this study, we have researched in vivo the regulation of DGK alpha, using a transgenic mouse model in which injection of an antigenic peptide activates the majority of peripheral T cells. We demonstrate that DGK alpha, highly expressed in resting T lymphocytes, is subject to complex control at the mRNA and protein levels during in vivo T cell activation. Subcellular fractionation of T lymphocytes shortly after in vivo engagement of the TCR shows rapid translocation of cytosolic DGK alpha to the membrane fraction. At early time points, DGK alpha translocation to the membrane correlates with rapid translocation of Ras guanyl nucleotide-releasing protein (RasGRP), a nucleotide exchange activator for Ras that associates to the membrane through a diacylglycerol-binding domain. To demonstrate a causal relationship between DGK alpha activity and RasGRP relocation to the membrane, we determined RasGRP translocation kinetics in a T cell line transiently transfected with constitutive active and dominant-negative DGK alpha mutants. We show that membrane localization of DGK alpha is associated with a negative regulatory signal for Ras activation by reversing RasGRP translocation. This study is the first demonstration of in vivo regulation of DGK alpha, and provides new insight into the functional role of a member of this family of lipid kinases in the regulation of the immune response. PMID- 12626539 TI - Detection of suppressor T lymphocytes and estimation of their frequency in limiting dilution assays by generalized linear regression modeling. AB - The estimate of the frequency of suppressor T lymphocytes in unfractionated cell populations remains challenging, mainly because these regulatory cells do not display specific immunophenotypic markers. In this paper, we describe a novel theoretical approach for quantifying the frequency of suppressor cells. This method is based on limiting dilution data modeling, and allows the simultaneous estimation of the frequencies of both proliferating and suppressor cells. We used previously published biological data, characterizing the inhibiting activity of suppressor T cell clones. Starting from these data, we propose a mathematical model describing the interaction between suppressor and proliferating T cells, and applied to a Poisson process. Limiting dilution data corresponding to this non-single-hit, suppressor two-target Poisson model were artificially generated, then modeled according to a generalized linear regression procedure. Deviation from the single-hit Poisson model was revealed by a statistical slope test, and a stepwise analysis of the regression appeared to be an efficient method that strongly argued in favor of the presence of suppressor cells. By using the frequency of proliferating T cells calculated in the first step of the regression, we demonstrated the possibility to provide a reasonable estimate of the frequency of suppressor T cells. Based on these findings, a practical decision-making procedure is given to perform standard analyses of limiting dilution data. PMID- 12626541 TI - Role of the cytoplasmic tails of CXCR1 and CXCR2 in mediating leukocyte migration, activation, and regulation. AB - IL-8 (or CXCL8) activates the receptors CXCR1 (IL-8RA) and CXCR2 (IL-8RB) to induce chemotaxis in leukocytes, but only CXCR1 mediates cytotoxic and cross regulatory signals. This may be due to the rapid internalization of CXCR2. To investigate the roles of the intracellular domains in receptor regulation, wild type, chimeric, phosphorylation-deficient, and cytoplasmic tail (C-tail) deletion mutants of both receptors were expressed in RBL-2H3 cells and studied for cellular activation, receptor phosphorylation, desensitization, and internalization. All but one chimeric receptor bound IL-8 and mediated signal transduction, chemotaxis, and exocytosis. Upon IL-8 activation, the chimeric receptors underwent receptor phosphorylation and desensitization. One was resistant to internalization, yet it mediated normal levels of beta-arrestin 2 (beta arr-2) translocation. The lack of internalization by this receptor may be due to its reduced association with beta arr-2 and the adaptor protein-2 beta. The C-tail-deleted and phosphorylation-deficient receptors were resistant to receptor phosphorylation, desensitization, arrestin translocation, and internalization. They also mediated greater phosphoinositide hydrolysis and exocytosis and sustained Ca(2+) mobilization, but diminished chemotaxis. These data indicate that phosphorylation of the C-tails of CXCR1 and CXCR2 are required for arrestin translocation and internalization, but are not sufficient to explain the rapid internalization of CXCR2 relative to CXCR1. The data also show that receptor internalization is not required for chemotaxis. The lack of receptor phosphorylation was correlated with greater signal transduction but diminished chemotaxis, indicating that second messenger production, not receptor internalization, negatively regulates chemotaxis. PMID- 12626540 TI - Vav-1 and the IKK alpha subunit of I kappa B kinase functionally associate to induce NF-kappa B activation in response to CD28 engagement. AB - We have recently observed that CD28 engagement initiates a signaling pathway leading to the activation of I kappa B kinase (IKK) complex and, consequently, to NF-kappa B activation, and we identified Vav-1 as an important mediator of this function. Here we report for the first time that Vav-1 constitutively associates with IKK alpha in both Jurkat and primary CD4(+) T cells. Vav-1/IKK alpha association is mediated by their helix-loop-helix domains, does not involve IKK beta, and is functionally relevant in that Vav-1-associated IKK alpha kinase activity is increased following CD28 engagement by B7. Moreover, we demonstrate that CD28-induced NF-kappa B activation is augmented by both IKK alpha and Vav-1, but not IKK beta. Confocal microscopy showed that endogenous Vav-1 and IKK alpha, but not IKK beta, were recruited to the membrane and colocalized in response to CD28 stimulation. Taken together, these data evidence that Vav-1 plays a key role in the control of NF-kappa B pathway by targeting IKK alpha in the T cell membrane and favoring its activation in response to CD28 stimulation. PMID- 12626542 TI - Provision of 4-1BB ligand enhances effector and memory CTL responses generated by immunization with dendritic cells expressing a human tumor-associated antigen. AB - Up-regulation of receptor-ligand pairs during interaction of an MHC-presented epitope on dendritic cells (DCs) with cognate TCR may amplify, sustain, and drive diversity in the ensuing T cell immune response. Members of the TNF ligand superfamily and the TNFR superfamily contribute to this costimulatory molecule signaling. In this study, we used replication deficient adenoviruses to introduce a model tumor-associated Ag (the E7 oncoprotein of human papillomavirus 16) and the T cell costimulatory molecule 4-1BBL into murine DCs, and monitored the ability of these recombinant DCs to elicit E7-directed T cell responses following immunization. Splenocytes from mice immunized with DCs expressing E7 alone elicited E7-directed effector and memory CTL responses. Coexpression of 4-1BBL in these E7-expressing DCs increased effector and memory CTL responses when they were used for immunization. 4-1BBL expression up-regulated CD80 and CD86 second signaling molecules in DCs. We also report an additive effect of 4-1BBL and receptor activator of NF-kappa B/receptor activator of NF-kappa B ligand coexpression in E7-transduced DC immunogens on E7-directed effector and memory CTL responses and on MHC class II and CD80/86 expression in DCs. Additionally, expression of 4-1BBL in E7-transduced DCs reduced nonspecific T cell activation characteristic of adenovirus vector-associated immunization. The results have generic implications for improved or tumor Ag-expressing DC vaccines by incorporation of exogenous 4-1BBL. There are also specific implications for an improved DC-based vaccine for human papillomavirus 16-associated cervical carcinoma. PMID- 12626543 TI - Acute alcohol inhibits TNF-alpha processing in human monocytes by inhibiting TNF/TNF-alpha-converting enzyme interactions in the cell membrane. AB - Alcohol abuse has long been known to adversely affect innate immune responses and predispose to infections. One cellular mechanism responsible for this effect is alcohol-induced suppression of TNF-alpha by mononuclear phagocytes. We undertook experiments to better understand the cellular mechanisms by which alcohol dose dependently suppresses TNF elaboration by human monocytes. Here we show in human primary monocytes and cell lines that alcohol suppresses LPS-induced TNF secretion post-transcriptionally by inhibiting cellular processing by TNF-alpha converting enzyme (TACE). Using fluorescent resonance energy transfer microscopy, physiological relevant levels of alcohol resulted in a reversible dose-dependent decrease in fluorescent resonance energy transfer efficiency between TNF and TACE. These data demonstrate that alcohol inhibits interactions between TNF and its converting enzyme, TACE, possibly by affecting membrane fluidity. These data in part explain the cellular mechanisms by which alcohol impairs monocyte function and may identify immunotherapeutic targets aimed at restoring immune function in this at-risk patient population. PMID- 12626544 TI - Glucocorticoids alter the lipid and protein composition of membrane rafts of a murine T cell hybridoma. AB - Glucocorticoids (GC) are widely used anti-inflammatory agents known to suppress T cell activation by interfering with the TCR activation cascade. The attenuation of early TCR signaling events by these compounds has been recently attributed to a selective displacement of key signaling proteins from membrane lipid rafts. In this study, we demonstrate that GC displace the acyl-bound adaptor proteins linker for activation of T cells and phosphoprotein associated with glycosphingolipid-enriched microdomains from lipid rafts of murine T cell hybridomas, possibly by inhibiting their palmitoylation status. Analysis of the lipid content of the membrane rafts revealed that GC treatment led to a significant decrease in palmitic acid content. Moreover, we found an overall decrease in the proportion of raft-associated saturated fatty acids. These changes were consistent with a decrease in fluorescence anisotropy of isolated lipid rafts, indicating an increase in their fluidity. These findings identify the mechanisms underlying the complex inhibitory effects of glucocorticoids on early TCR signaling and suggest that some of the inhibitory properties of GC on T cell responses may be related to their ability to affect the membrane lipid composition and the palmitoylation status of important signaling molecules. PMID- 12626545 TI - A pool of central memory-like CD4 T cells contains effector memory precursors. AB - The L51S mutation in the D10.G4.1 TCR alpha-chain reduces the affinity of the TCR to its ligand by affecting the interactions among the TCR, the beta-chain of I A(k), and the bound peptide. We show that this mutation drives the generation of a pool of memory CD44(high)CD62L(neg)CD45RB(neg) CD4 TCR transgenic T cells. Their activation threshold is low, such that they proliferate in response to lower concentrations of agonist peptides than naive L51S CD4 T cells. Unlike effector memory CD4 T cells, however, they lack immediate effector function in response to TCR stimulation. These cells express IL-2R alpha only after culture with specific peptide. Although they can be recovered from lymph nodes, the majority lack the expression of the lymph node homing receptor CCR7. When these cells receive a second TCR stimulation in vitro, they differentiate into potent Th2-like effector cells, producing high levels of IL-4 at doses of agonist peptide too low to stimulate cytokine release from similarly differentiated naive L51S CD4 T cells. Having these properties, the L51S TCR transgenic memory CD4 T cells cannot be classified as either strict central memory or effector memory, but, rather, as a pool of memory T cells containing effector memory precursors. PMID- 12626547 TI - Protein kinase A regulates GATA-3-dependent activation of IL-5 gene expression in Th2 cells. AB - Treatment of Th cells with compounds that elevate cAMP levels augments Th2-type lymphokine expression, in particular the synthesis of IL-5. Using primary murine CD4(+) T lymphocytes, we show in this study that inhibition of protein kinase A (PKA) activity in Th2 effector cells impairs IL-5 synthesis, whereas the expression of PKA catalytic subunit alpha enhances IL-5 synthesis in Th0 cells. In addition, we observed by coexpression of PKA catalytic subunit and GATA-3 in Th1 cells that the stimulatory effect of PKA is dependent on GATA-3 activity. These data demonstrate that activation of PKA in Th effector cells induces the IL 5 gene expression in a GATA-3-dependent manner. PMID- 12626546 TI - The role of the CD134-CD134 ligand costimulatory pathway in alloimmune responses in vivo. AB - The CD134-CD134 ligand (CD134L) costimulatory pathway has been shown to be critical for both T and B cell activation; however, its role in regulating the alloimmune response remains unexplored. Furthermore, its interactions with other costimulatory pathways and immunosuppressive agents are unclear. We investigated the effect of CD134-CD134L pathway blockade on allograft rejection in fully MHC mismatched rat cardiac and skin transplantation models. CD134L blockade alone did not prolong graft survival compared with that of untreated recipients, and in combination with donor-specific transfusion, cyclosporine, or rapamycin, was less effective than B7 blockade in prolonging allograft survival. However, in combination with B7 blockade, long-term allograft survival was achieved in all recipients (>200 days). Moreover, this was synergistic in reducing the frequency of IFN-gamma-producing alloreactive lymphocytes and inhibiting the generation of activated/effector lymphocytes. Most impressively, this combination prevented rejection in a presensitized model using adoptive transfer of primed lymphocytes into athymic heart transplant recipients. In comparison to untreated recipients (mean survival time (MST): 5.3 +/- 0.5 days), anti-CD134L mAb alone modestly prolonged allograft survival (MST: 14 +/- 2.8 days) as did CTLA4Ig (MST: 21.5 +/- 1.7 days), but all grafts were rejected within 24 days. Importantly, combined blockade further and significantly prolonged allograft survival (MST: 75.3 +/- 12.7 days) and prevented the expansion and/or persistence of primed/effector alloreactive T cells. Our data suggest that CD134-CD134L is a critical pathway in alloimmune responses, especially recall/primed responses, and is synergistic with CD28-B7 in mediating T cell effector responses during allograft rejection. Understanding the mechanisms of collaboration between these different pathways is important for the development of novel strategies to promote long-term allograft survival. PMID- 12626548 TI - Secretion of IL-2 and IFN-gamma, but not IL-4, by antigen-specific T cells requires extracellular ATP. AB - Extracellular ATP and other nucleotides transmit signals to cells via surface associated molecules whose binding sites face the extracellular milieu. Ecto nucleoside triphosphate diphosphohydrolase is such an ATP-binding enzyme that is expressed by activated lymphocytes. We have previously shown that nonhydrolyzable ATP analogs block the lytic activity of NK cells and CD8(+) T cells as well as their E-NTPDase activity. These results suggest that the hydrolysis of ATP may play a role in lymphocyte function. Here we report that E-NTPDase activity is up regulated within 15 min of T cell stimulation and that reversible and irreversible enzyme inhibitors profoundly reduce secretion of IL-2 and IFN-gamma, but not IL-4. TNF-alpha, IL-10, and IL-5 production showed intermediate sensitivity to these ATP analogs. Depletion of extracellular ATP also inhibited secretion of IFN-gamma, but not IL-4, supporting the interpretation that extracellular ATP is required for secretion of some, but not all, cytokines. E NTPDase antagonists reduced transcription of IL-2 mRNA and inhibited TCR-mediated intracellular calcium flux. These results suggest that extracellular ATP plays an essential role in the TCR-mediated signal transduction cascade for expression of certain cytokine genes. PMID- 12626549 TI - The cyclic adenosine 5'-monophosphate response element modulator suppresses IL-2 production in stimulated T cells by a chromatin-dependent mechanism. AB - The production of IL-2 is tightly controlled by several transcription factors that bind to the IL-2 promoter. The cAMP response element modulator (CREM) is known to form complexes with CREB and bind to the -180 site of the IL-2 promoter in anergic and in systemic lupus erythematosus T cells. In this study we show that CREM is transcriptionally induced in T cells following stimulation through CD3 and CD28, binds to the IL-2 promoter in vivo, and suppresses IL-2 production. Transfection of an antisense CREM plasmid into T cells blocked the expression and binding of CREM to the IL-2 promoter and the decrease of IL-2 production, which follows the early increase after T cell stimulation with CD3 and CD28. In addition, as assessed by chromatin immunoprecipitation experiments, antisense CREM prevented the binding of protein 300 and cAMP response element binding protein and promoted the acetylation of histones. Antisense CREM also enhanced the accessibility of the IL-2 promoter to endonucleases and prevented the condensation of chromatin in vivo. Our data suggest that upon T cell activation, CREM gradually replaces phosphorylated CREB at the -180 site of the IL-2 promoter. CREM, in turn, binds protein 300 and cAMP response element binding protein, but CREM is unable to activate its histone acetyltransferase activity, which results in condensation of chromatin and down-regulation of IL-2 production. PMID- 12626551 TI - Regulatory T cells control autoimmunity in vivo by inducing apoptotic depletion of activated pathogenic lymphocytes. AB - Clinical autoimmunity requires both activation of self-reactive T cells as well as a failure of peripheral tolerance mechanisms. We previously identified one such mechanism that involves regulatory T cells recognizing TCR V beta 8.2 chain derived peptides in the context of MHC. How this regulation affects the fate of target V beta 8.2(+) T lymphocytes in vivo that mediate experimental autoimmune encephalomyelitis has remained unknown. The present study using immunoscope and CFSE-labeling analysis demonstrates that the expansion of regulatory CD4 and CD8 T cells in vivo results in apoptotic depletion of the dominant, myelin basic protein-reactive V beta 8.2(+) T cells, but not subdominant V beta 13(+) T cells. The elimination of only activated T cells by this negative feedback mechanism preserves the remainder of the naive V beta 8.2(+) T cell repertoire and at the same time results in protection from disease. These studies are the first in clearly elucidating the fate of myelin basic protein-specific encephalitogenic T cells in vivo following regulation. PMID- 12626550 TI - Quantifying recruitment of cytosolic peptides for HLA class I presentation: impact of TAP transport. AB - MHC class I ligands are recruited from the cytosolic peptide pool, whose size is likely to depend on the balance between peptide generation by the proteasome and peptide degradation by downstream peptidases. We asked what fraction of this pool is available for presentation, and how the size of this fraction is modulated by peptide affinity for the TAP transporters. A model epitope restricted by HLA-A2 and a series of epitope precursors with N-terminal extensions by single residues modifying TAP affinity were expressed in a system that allowed us to monitor and modulate cytosolic peptide copy numbers. We show that presentation varies strongly according to TAP affinities of the epitope precursors. The fraction of cytosolic peptides recruited for MHC presentation does not exceed 1% and is more than two logs lower for peptides with very low TAP affinities. Therefore, TAP affinity has a substantial impact on MHC class I Ag presentation. PMID- 12626552 TI - Cu/Zn superoxide dismutase plays important role in immune response. AB - Activation of macrophages leads to the secretion of cytokines and enzymes that shape the inflammatory response and increase metabolic processes. This, in turn, results in increased production of reactive oxygen species. The role of Cu/Zn superoxide dismutase (SOD-1), an important enzyme in cellular oxygen metabolism, was examined in activated peritoneal elicited macrophages (PEM) and in several inflammatory processes in vivo. LPS and TNF-alpha induced SOD-1 in PEM. SOD-1 induction by LPS was mainly via extracellular signal-regulated kinase-1 activation. Transgenic mice overexpressing SOD-1 demonstrated a significant increase in the release of TNF-alpha and of the metalloproteinases MMP-2 and MMP 9 from PEM. Disulfiram (DSF), an inhibitor of SOD-1, strongly inhibited the release of TNF-alpha, vascular endothelial growth factor, and MMP-2 and MMP-9 from cultured activated PEM. These effects were prevented by addition of antioxidants, further indicating involvement of reactive oxygen species. In vivo, transgenic mice overexpressing SOD-1 demonstrated a 4-fold increase in serum TNF alpha levels and 2-fold stronger delayed-type hypersensitivity reaction as compared with control nontransgenic mice. Conversely, oral administration of DSF lowered TNF-alpha serum level by 4-fold, lowered the delayed-type hypersensitivity response in a dose-dependent manner, and significantly inhibited adjuvant arthritis in Lewis rats. The data suggest an important role for SOD-1 in inflammation, establish DSF as a potential inhibitor of inflammation, and raise the possibility that regulation of SOD-1 activity may be important in the treatment of immune-dependent pathologies. PMID- 12626553 TI - CD28 signal enhances apoptosis of CD8 T cells after strong TCR ligation. AB - High avidity ligation of the TCR induces negative selection in the thymus and can also induce apoptosis of peripheral T cells. Costimulation through CD28 enhances T cell activation and facilitates negative selection in the thymus, but the role of CD28 in peripheral T cell deletional tolerance has not been investigated. We used 2C CD28 wild-type and 2C CD28-deficient strains to assess the effects of CD28 and TCR avidity on peripheral T cell expansion and apoptosis. We compared the activation, division, expansion, and apoptosis of CD28(+/+) and CD28(-/-) 2C cells in response to self-Ag (K(b)), alloantigens with intermediate (K(bm3)), high (L(d)), or very high (L(d) + QL9 peptide) avidity. With intermediate avidity alloantigen, the CD28 signal enhanced T cell activation and expansion. However, when T cells encountered high avidity alloantigen, the CD28 signal reduced T cell expansion and increased apoptosis. These results indicate that the CD28 signal can down-regulate peripheral T cell responses by increasing apoptosis when TCR ligation exceeds a critical threshold. PMID- 12626554 TI - A threshold for central T cell tolerance to an inducible serum protein. AB - We report an inducible system of self Ag expression that examines the relationship between serum protein levels and central T cell tolerance. This transgenic approach is based on tetracycline-regulated expression of a secreted form of hen egg lysozyme, tagged with a murine hemoglobin (Hb) epitope. In the absence of the tetracycline-regulated transactivator, serum levels of the chimeric protein are extremely low (< or = 0.1 ng/ml) and the mice show partial tolerance to both Hb(64-76) and lysozyme epitopes. In the presence of the transactivator, expression increases to 1.5 ng/ml and the mice are completely tolerant. Partial tolerance was further investigated by crossing these mice to strains expressing transgenic TCRs. At the lowest Ag levels, 3.L2tg T cells (specific for Hb(64-76)/I-E(k)) escape the thymus and approximately 10% of CD4(+) splenocytes express the 3.L2 TCR. In contrast, 3A9 T cells (specific for hen egg lysozyme(46-61)/I-A(k)) are completely eliminated by negative selection. These data define a tolerogenic threshold for negative selection of Ag-specific T cells by circulating self proteins that are 100-fold more sensitive than previously demonstrated. They suggest that partial tolerance at extremely low levels of self Ag exposure is the result of a restricted repertoire of responding T cells, rather than a simple reduction in precursor frequency; tolerogenic thresholds are T cell specific. PMID- 12626555 TI - Blockade of CD40-mediated signaling is sufficient for inducing islet but not skin transplantation tolerance. AB - Treatment of mice with a single donor-specific transfusion (DST) plus a brief course of anti-CD154 mAb to block CD40-mediated signaling uniformly induces donor specific transplantation tolerance. Survival of islet allografts in treated mice is permanent, but skin grafts eventually fail unless recipients are thymectomized. The nature of the cellular mechanisms involved and the basis for the difference in survival of islet vs skin allografts are not known. In this study, we used CD40 knockout mice to investigate the role of CD40-mediated signaling in each component of the tolerance induction protocol: the DST, the graft, and the host. When CD40-mediated signaling was eliminated in only the DST or the graft, islet allografts were rapidly rejected. However, when CD40 signaling was eliminated in the host, approximately 40% of the islet allografts survived. When CD40 signaling was eliminated in the DST, the graft, and the host, islet grafts survived long term (>84 days), whereas skin allografts were rapidly rejected ( approximately 13 days). We conclude that transplantation tolerance induction in mice treated with DST and anti-CD154 mAb requires blockade of CD40 mediated signaling in the DST, the graft, and the host. Blockade of CD40-mediated signaling is necessary and sufficient for inducing islet allograft tolerance and is necessary but not sufficient for long-term skin allograft survival. We speculate that a requirement for regulatory CD4(+) T cells in skin allograft recipients could account for this differential response to tolerance induction. PMID- 12626556 TI - Activation of alloreactive CD8+ T cells operates via CD4-dependent and CD4 independent mechanisms and is CD154 blockade sensitive. AB - CD154, one of the most extensively studied T cell costimulation molecules, represents a promising therapeutic target in organ transplantation. However, the immunological mechanisms of CD154 blockade that result in allograft protection, particularly in the context of alloreactive CD4/CD8 T cell activation, remain to be elucidated. We now report on the profound inhibition of alloreactive CD8(+) T cells by CD154 blockade via both CD4-dependent and CD4-independent activation pathways. Using CD154 KO recipients that are defective in alloreactive CD8(+) T cell activation and unable to reject cardiac allografts, we were able to restore CD8 activation and graft rejection by adoptively transferring CD4(+) or CD8(+) T cells from wild-type syngeneic donor mice. CD4-independent activation of alloreactive CD8(+) T cells was confirmed following treatment of wild-type recipients with CD4-depleting mAb, and by using CD4 KO mice. Comparable levels of alloreactive CD8(+) T cell activation was induced by allogenic skin engraftment in both animal groups. CD154 blockade inhibited CD4-independent alloreactive CD8(+) T cell activation. Furthermore, we analyzed whether disruption of CD154 signaling affects cardiac allograft survival in skin-sensitized CD4 KO and CD8 KO recipients. A better survival rate was observed consistently in CD4 KO, as compared with CD8 KO recipients. Our results document CD4-dependent and CD4 independent activation pathways for alloreactive CD8(+) T cells that are both sensitive to CD154 blockade. Indeed, CD154 blockade was effective in preventing CD8(+) T cell-mediated cardiac allograft rejection. PMID- 12626557 TI - Ca2+ stores and Ca2+ entry differentially contribute to the release of IL-1 beta and IL-1 alpha from murine macrophages. AB - Interleukin-1 is a primary mediator of immune responses to injury and infection, but the mechanism of its cellular release is unknown. IL-1 exists as two agonist forms (IL-1 alpha and IL-1 beta) present in the cytosol of activated monocytes/macrophages. IL-1 beta is synthesized as an inactive precursor that lacks a signal sequence, and its trafficking does not use the classical endoplasmic reticulum-Golgi route of secretion. Using primary cultured murine peritoneal macrophages, we demonstrate that P2X7 receptor activation causes release of IL-1 beta and IL-1 alpha via a common pathway, dependent upon the release of Ca(2+) from endoplasmic reticulum stores and caspase-1 activity. Increases in intracellular Ca(2+) alone do not promote IL-1 secretion because a concomitant efflux of K(+) through the plasmalemma is required. In addition, we demonstrate the existence of an alternative pathway for the secretion of IL-1 alpha, independent of P2X7 receptor activation, but dependent upon Ca(2+) influx. The identification of these mechanisms provides insight into the mechanism of IL 1 secretion, and may lead to the identification of targets for the therapeutic modulation of IL-1 action in inflammation. PMID- 12626558 TI - Mast cell-derived exosomes induce phenotypic and functional maturation of dendritic cells and elicit specific immune responses in vivo. AB - Mast cells (MCs) are considered major players in IgE-mediated allergic responses, but have also recently been recognized as active participants in innate as well as specific immune responses. Recent work provided evidence that MCs are able to activate B and T lymphocytes through the release of vesicles called exosomes. Here we demonstrate that exosomes, which are located in the endocytic pathway, harbor exogenous Ags that associate with other molecules endowed with immunomodulatory functions, including 60- and 70-kDa heat shock proteins. Administration to naive mice of Ag-containing exosomes in the absence of conventional adjuvants elicits specific Ab responses across the MHC II haplotype barrier. We demonstrate that MC-exosomes induce immature dendritic cells (DCs) to up-regulate MHC class II, CD80, CD86, and CD40 molecules and to acquire potent Ag presenting capacity to T cells. Uptake and processing of Ag-associated exosomes by endogenous DCs were also demonstrated. Finally, exosome-associated heat shock proteins are critical for the acquisition by DCs of the Ag-presenting function. This work demonstrates a heretofore unrecognized collaborative interaction between MCs and DCs leading to the elicitation of specific immune responses. PMID- 12626559 TI - Elimination of leukemia in the absence of lethal graft-versus-host disease after allogenic bone marrow transplantation. AB - Donor T cells are able to effect a graft-vs-leukemia (GVL) response but also induce graft-vs-host disease (GVHD) after allogeneic bone marrow transplantation. We used an AKR leukemia murine transplant model, analogous to human acute lymphoblastic leukemia, in which donor T cells expressed a thymidine kinase suicide gene, to test whether separation of GVL and graft-vs-host (GVH) responses was feasible by selectively eliminating alloactivated donor T cells at defined time points posttransplant. Under experimental conditions where untreated mice could not be cured of disease without dying from GVHD, mice transplanted with thymidine kinase-positive T cells and subsequently administered ganciclovir (GCV) could eliminate leukemia without lethal GVHD. Timing of GCV administration, donor T cell dose, and preexisting leukemia burden were observed to be critical variables. Eradication of leukemia without lethal GVHD in GCV-treated mice implied that the kinetics of GVL and GVH responses were asynchronous and could therefore be temporally dissociated by timely GCV administration. That this strategy was feasible in a murine leukemia model in which GVHD and GVL reactivity are tightly linked suggests that this approach may be relevant to the treatment of selected human leukemias where similar constraints exist. This strategy represents an alternative approach to separating GVL and GVH reactivity and challenges the current paradigm that separation of these responses is dependent upon the administration of donor T cells with restricted specificity for leukemia as opposed to host Ags. PMID- 12626560 TI - IgE-mediated activation of NK cells through Fc gamma RIII. AB - NK cells express Fc gamma RIII (CD16), which is responsible for IgG-dependent cell cytotoxicity and for production of several cytokines and chemokines. Whereas Fc gamma RIII on NK cells is composed of both Fc gamma RIII alpha and FcR gamma chains, that on mast cells is distinct from NK cells and made of Fc gamma RIII alpha, FcR beta, and FcR gamma. Mast cells show degranulation and release several mediators, which cause anaphylactic responses upon cross-linking of Fc gamma RIII as well as Fc epsilon RI with aggregated IgE. In this paper, we examined whether IgE activates NK cells through Fc gamma RIII on their cell surface. We found that NK cells produce several cytokines and chemokines related to an allergic reaction upon IgE stimulation. Furthermore, NK cells exhibited cytotoxicity against IgE coated target cells in an Fc gamma RIII-dependent manner. These effects of IgE through Fc gamma RIII were not observed in NK cells from FcR gamma-deficient mice lacking Fc gamma RIII expression. Collectively, these results demonstrate that NK cells can be activated with IgE through Fc gamma RIII and exhibit both cytokine/chemokine production and Ab-dependent cell cytotoxicity. These data imply that not only mast cells but also NK cells may contribute to IgE-mediated allergic responses. PMID- 12626561 TI - The roles of Toll-like receptor 9, MyD88, and DNA-dependent protein kinase catalytic subunit in the effects of two distinct CpG DNAs on dendritic cell subsets. AB - Oligodeoxynucleotides containing unmethylated CpG motifs (CpG DNAs) can function as powerful immune adjuvants by activating APC. Compared with conventional phosphorothioate-backbone CpG DNAs, another type of CpG DNAs, called an A or D type (A/D-type), possesses higher ability to induce IFN-alpha production. Conventional CpG DNAs can exert their activity through Toll-like receptor 9 (TLR9) signaling, which depends on a cytoplasmic adapter, MyD88. However, it remains unknown how A/D-type CpG DNAs exhibit their immunostimulatory function. In this study we have investigated murine dendritic cell (DC) responses to these two distinct CpG DNAs. Not only splenic, but also in vitro bone marrow-derived, DCs could produce larger amounts of IFN-alpha in response to A/D-type CpG DNAs compared with conventional CpG DNAs. This IFN-alpha production was mainly due to the B220(+) DC subset. On the other hand, the B220(-) DC subset responded similarly to both CpG DNAs in terms of costimulatory molecule up-regulation and IL-12 induction. IFN-alpha, but not IL-12, induction was dependent on type I IFN. However, all activities of both CpG DNAs were abolished in TLR9- and MyD88-, but were retained in DNA-PKcs-deficient DCs. This study demonstrates that the TLR9 MyD88 signaling pathway is essential for all DC responses to both types of CpG DNAs. PMID- 12626562 TI - Proline-rich tyrosine kinase 2 and Rac activation by chemokine and integrin receptors controls NK cell transendothelial migration. AB - Protein tyrosine kinase activation is an important requisite for leukocyte migration. Herein we demonstrate that NK cell binding to endothelium activates proline-rich tyrosine kinase 2 (Pyk-2) and the small GTP binding protein Rac that are coupled to integrin and chemokine receptors. Chemokine-mediated, but not integrin-mediated, Pyk-2 and Rac activation was sensitive to pretreatment of NK cells with pertussis toxin, a pharmacological inhibitor of G(i) protein-coupled receptors. Both Pyk-2 and Rac are functionally involved in chemokine-induced NK cell migration through endothelium or ICAM-1 or VCAM-1 adhesive proteins, as shown by the use of recombinant vaccinia viruses encoding dominant negative mutants of Pyk-2 and Rac. Moreover, we found that Pyk-2 is associated with the Rac guanine nucleotide exchange factor Vav, which undergoes tyrosine phosphorylation upon integrin triggering. Finally, we provide direct evidence for the involvement of Pyk-2 in the control of both chemokine- and integrin-mediated Rac activation. Collectively, our results indicate that Pyk-2 acts as a receptor proximal link between integrin and chemokine receptor signaling, and the Pyk 2/Rac pathway plays a pivotal role in the control of NK cell transendothelial migration. PMID- 12626564 TI - The four TCR genes of teleost fish: the cDNA and genomic DNA analysis of Japanese flounder (Paralichthys olivaceus) TCR alpha-, beta-, gamma-, and delta-chains. AB - We have isolated and identified all four TCR alpha, beta, gamma, and delta cDNAs and genomic clones from a Japanese flounder leukocyte cDNA library and bacterial artificial chromosomal genomic library. Numerous TCR transcripts were sequenced to examine the variability against antigenic peptide, and were shown hypervariability on their complementarity-determining region 3 (CDR3) loops. Among CDR3s, CDR3 delta showed a long and broad length distribution, indicating greater similarity to that of Ig. From cDNA sequences and genomic gene analysis of each chain, we found that flounder TCR beta, gamma, and delta have two different C gene segments, while the TCR alpha C region exists as a single segment. The flounder C gammas and C deltas showed different lengths in the connecting peptide (CP) region between the different types of polypeptides. The C delta 1 gene consists of two exons, one that encodes an extracellular Ig-like domain (exon 1) and the other that encodes either a very short or possibly a lacking CP region, a transmembrane region, and a cytoplasmic tail (exon 2); these are located within TCR alpha gene locus. Southern blot analysis, using the bacterial artificial chromosomal genomic DNA clones, revealed that the C delta 2 gene segment, which has a long CP region and different genomic organization to the C delta 1 gene, exists on same gene locus as the TCR gamma-chain. This suggests that the flounder possesses very unique genomic DNA organization and gene loci for TCR, C alpha/C delta 1, and C gamma/C delta 2. PMID- 12626563 TI - Differential distribution of HLA-DQ beta/DR beta epitopes in the two forms of Guillain-Barre syndrome, acute motor axonal neuropathy and acute inflammatory demyelinating polyneuropathy (AIDP): identification of DQ beta epitopes associated with susceptibility to and protection from AIDP. AB - Guillain-Barre syndrome (GBS), an acute, immune-mediated paralytic disorder affecting the peripheral nervous system, is the most common cause of acute flaccid paralysis in the post-polio era. GBS is classified into several subtypes based on clinical and pathologic criteria, with acute inflammatory demyelinating polyneuropathy (AIDP) and acute motor axonal neuropathy (AMAN) being the most common forms observed. To better understand the pathogenesis of GBS and host susceptibility to developing the disease, the distribution of HLA class II Ags along with the seroreactivity to Campylobacter jejuni were investigated in a population of GBS patients from northern China. Using DNA-based typing methods, 47 patients with AMAN, 25 patients with AIDP, and 97 healthy controls were studied for the distribution of class II alleles. We found that the DQ beta RLD(55-57)/ED(70-71) and DR beta E(9)V(11)H(13) epitopes were associated with susceptibility to AIDP (p = 0.009 and p = 0.004, respectively), and the DQ beta RPD(55-57) epitope was associated with protection (p = 0.05) from AIDP. These DQ beta/DR beta positional residues are a part of pockets 4 (DQ beta 70, 71, DR beta 13), 6 (DR beta 11), and 9 (DQ beta 56, 57, DR beta 9); have been demonstrated to be important in peptide binding and T cell recognition; and are associated with other diseases that have a pathoimmunological basis. Class II HLA associations were not identified with AMAN, suggesting a different immunological mechanism of disease induction in the two forms of GBS. These findings provide immunogenetic evidence for differentiating the two disease entities (AMAN and AIDP) and focuses our attention on particular DR beta/DQ beta residues that may be instrumental in understanding the pathophysiology of AIDP. PMID- 12626565 TI - Ikaros isoform x is selectively expressed in myeloid differentiation. AB - The Ikaros gene is alternately spliced to generate multiple DNA-binding and nonbinding isoforms that have been implicated as regulators of hematopoiesis, particularly in the lymphoid lineages. Although early reports of Ikaros mutant mice focused on lymphoid defects, these mice also show significant myeloid, erythroid, and stem cell defects. However, the specific Ikaros proteins expressed in these cells have not been determined. We recently described Ikaros-x (Ikx), a new Ikaros isoform that is the predominant Ikaros protein in normal human hematopoietic cells. In this study, we report that the Ikx protein is selectively expressed in human myeloid lineage cells, while Ik1 predominates in the lymphoid and erythroid lineages. Both Ik1 and Ikx proteins are expressed in early human hematopoietic cells (Lin(-)CD34(+)). Under culture conditions that promote specific lineage differentiation, Ikx is up-regulated during myeloid differentiation but down-regulated during lymphoid differentiation from human Lin(-)CD34(+) cells. We show that Ikx and other novel Ikaros splice variants identified in human studies are also expressed in murine bone marrow. In mice, as in humans, the Ikx protein is selectively expressed in the myeloid lineage. Our studies suggest that Ikaros proteins function in myeloid, as well as lymphoid, differentiation and that specific Ikaros isoforms may play a role in regulating lineage commitment decisions in mice and humans. PMID- 12626566 TI - CD40-mediated transcriptional regulation of the IL-6 gene in B lymphocytes: involvement of NF-kappa B, AP-1, and C/EBP. AB - Engagement of CD40 by its ligand CD154 induces IL-6 production by B lymphocytes. We previously reported that this IL-6 production is dependent upon binding of the adapter protein TNF receptor-associated factor 6 to the cytoplasmic domain of CD40, while binding of TNF receptor-associated factors 2 and 3 is dispensable, as is the activation-induced nuclear translocation of NF-kappa B. The present study was designed to characterize CD40-mediated transcriptional control of the IL-6 gene in B cells. CD40 engagement on B lymphocytes activated the IL-6 promoter, and mutations in the putative binding sites for AP-1 and C/EBP transcription factors reduced this activation. Interestingly, a mutation in the putative NF kappa B binding site completely abrogated the basal promoter activity, thus also rendering the promoter unresponsive to CD40 stimulation, suggesting that this site is required for binding of NF-kappa B constitutively present in the nucleus of mature B cells. The expression of dominant negative Fos or C/EBP alpha proteins, which prevent binding of AP-1 or C/EBP complexes to DNA, also reduced CD40-mediated IL-6 gene expression. Furthermore, CD40 stimulation led to phosphorylation of c-Jun on its activation domain, implicating CD40-mediated Jun kinase activation in the transcriptional regulation of IL-6 production. PMID- 12626567 TI - Role of NFATx (NFAT4/NFATc3) in expression of immunoregulatory genes in murine peripheral CD4+ T cells. AB - Ca(2+)-regulated NFAT family members are transcription factors crucial for the expression of various cytokine genes and other immunoregulatory genes. Analyses of mice defective in one or two NFAT family members have revealed functions specific to each NFAT gene. However, the redundant functions of several family members limit the usefulness of gene disruption analysis. For example, CD4(+) T cells isolated from NFATx-disrupted mice do not show any modulation in cytokine gene expression, perhaps because other family members compensate for its absence. To analyze the role of NFATx in the regulation of immunoregulatory genes in T cells, we made a gain-of-function mutant by creating transgenic mice expressing a constitutively nuclear form of NFATx in T cell lineages. In naive CD4(+) T cells, NFATx up-regulated the expression of several cytokine genes and activation markers and suppressed the expression of CD154. In Th1 cells, NFATx enhanced the expression of the Th1 cytokine genes, IFN-gamma and TNF-alpha. In contrast, NFATx suppressed Th2 cytokine genes such as IL-4 and IL-5 in Th2 cells. It has been reported that both NFAT1 and NFATx are required to maintain the homeostasis of the immune system. Our results suggest that NFATx exerts this function by inhibiting the expression of some critical immunoregulatory genes. PMID- 12626568 TI - NF-kappa B is required for surface Ig-induced Fas resistance in B cells. AB - The susceptibility of primary murine B cells to Fas-mediated apoptosis is regulated in a receptor-specific fashion. Whereas CD40 engagement produces marked sensitivity to Fas killing, engagement of the B cell Ag receptor blocks Fas signaling for cell death in otherwise Fas-sensitive, CD40-stimulated targets and thus induces Fas resistance. The signaling pathway that leads from B cell Ag receptor to Fas resistance has not been fully characterized, but has been shown to depend on new gene expression. NF-kappa B is activated following B cell Ag receptor engagement and is associated with antiapoptosis; thus, it would seem a likely candidate to mediate transcriptional activation for inducible Fas resistance. Inhibition of B cell Ag receptor signaling for NF-kappa B activation completely blocked induction of Fas resistance by anti-Ig, and this same phenotype was observed both with chemical inhibitors such as lactacystin and pyrrolidinedithiocarbamate as well as with an I kappa B alpha dominant negative TAT fusion protein. Antiapoptotic, NF-kappa B-responsive transcripts include two gene products previously implicated in mediating anti-Ig-induced Fas resistance, Bcl-x(L) and FLIP. B cell Ag receptor-induced up-regulation of both these gene products was blocked by NF-kappa B inhibition, suggesting a mechanism by which the loss of nuclear NF-kappa B alters the sensitivity of B cell Ag receptor stimulated B cells to Fas-mediated apoptosis. These results indicate that activation of NF-kappa B plays a key role in mediating Fas resistance produced by B cell Ag receptor engagement. PMID- 12626569 TI - Identification of a functionally impaired positive regulatory domain I binding factor 1 transcription repressor in myeloma cell lines. AB - B cell differentiation into a plasma cell requires expression of the positive regulatory domain zinc finger protein 1 gene (PRDM1) that encodes the positive regulatory domain I binding factor 1 (PRDI-BF1 or Blimp-1) protein. It represses the transcription of specific target genes, including c-myc, the MHC class II trans-activator, Pax-5, and CD23b. In this study we demonstrate the presence of an alternative protein product of the PRDM1 gene. The new protein, PRDI-BF1 beta, has a disrupted PR domain and lacks the amino-terminal 101 aa of the originally described protein. PRDI-BF1 beta has a dramatic loss of repressive function on multiple target genes, but maintains normal DNA-binding activity, nuclear localization, and association with histone deacetylases and deacetylase activity. Myeloma cell lines express the highest levels of PRDM1 beta mRNA relative to the full-length form, while primary cells and several other cell lines have very low, but detectable, levels of PRDM1 beta. RNA analysis and analysis of the PRDM1 promoters demonstrate that PRDI-BF1 beta is generated from the same gene by alternative transcription initiation using an internal promoter. These newly described features of the PRDM1 gene are highly analogous to the PRDM2 (RIZ) and PRDM3 (MDS1-EVI1) genes, in which each express a truncated protein missing the PR domain. The expression of each of the truncated proteins is elevated in cancerous cells and may play an important role in the disease. PMID- 12626570 TI - Human IgG2 can form covalent dimers. AB - Unlike IgA and IgM, IgG has not yet been shown to form covalent polymers. However in the presence of specific Ag, murine IgG3 has been shown to polymerize through noncovalent interactions. In contrast to the noncovalent oligomers found with murine IgG3, we have detected covalent dimers in three different recombinant human IgG2 Abs produced in myeloma cells. Both IgG2,kappa and IgG2,lambda can form dimers. In addition, analysis of pooled human gamma globulin and several normal sera revealed the presence of IgG2 dimers. The IgG2 dimers are in contrast to the noncovalent IgG dimers found in pooled sera of multiple donors resulting from idiotype/anti-idiotype (Id/anti-Id) interactions. Cyanogen bromide cleavage analysis suggests that one or more Cys residues in the gamma 2 hinge are involved in dimer assembly. The potential role of IgG2 dimers in immunity against carbohydrate Ags is discussed. PMID- 12626572 TI - Role of CD1d in coxsackievirus B3-induced myocarditis. AB - The myocarditic (H3) variant of Coxsackievirus B3 (CVB3) causes severe myocarditis in BALB/c mice and BALB/c mice lacking the invariant J alpha 281 gene, but minimal disease in BALB/c CD1d(-/-) animals. This indicates that CD1d expression is important in this disease but does not involve the invariant NKT cell often associated with CD1d-restricted immunity. The H3 variant of the virus increases CD1d expression in vitro in neonatal cardiac myocytes whereas a nonmyocarditic (H310A1) variant does not. V gamma 4(+) T cells show increased activation in both H3-infected BALB/c and J alpha 281(-/-) mice compared with CD1d(-/-) animals. The activated BALB/c V gamma 4(+) T cells from H3-infected mice kill H3-infected BALB/c myocytes and cytotoxicity is blocked with anti-CD1d but not with anti-MHC class I (K(d)/D(d)) or class II (IA/IE) mAbs. In contrast, H3 virus-infected CD1d(-/-) myocytes are not killed. These studies demonstrate that CD1d expression is essential for pathogenicity of CVB3-induced myocarditis, that CD1d expression is increased early after infection in vivo in CD1d(+) mice infected with the myocarditic but not with the nonmyocarditic CVB3 variant, and that V gamma 4(+) T cells, which are known to promote myocarditis susceptibility, appear to recognize CD1d expressed by CVB3-infected myocytes. PMID- 12626571 TI - Inhibition of NF-kappa B activity in T and NK cells results in defective effector cell expansion and production of IFN-gamma required for resistance to Toxoplasma gondii. AB - To define the role of NF-kappa B in the development of T cell responses required for resistance to Toxoplasma gondii, mice in which T cells are transgenic for a degradation-resistant (Delta N) form of I kappa B alpha, an inhibitor of NF-kappa B, were challenged with T. gondii and their response to infection compared with control mice. I kappa B alpha(Delta N)-transgenic (Tg) mice succumbed to T. gondii infection between days 12 and 35, and death was associated with an increased parasite burden compared with wild-type (Wt) controls. Analysis of the responses of infected mice revealed that IL-12 responses were comparable between strains, but Tg mice had a marked reduction in systemic levels of IFN-gamma, the major mediator of resistance to T. gondii. In addition, the infection-induced increase in NK cell activity observed in Wt mice was absent from Tg mice and this correlated with NK cell expression of the transgene. Infection-induced activation of CD4(+) T cells was similar in Wt and Tg mice, but expansion of activated CD4(+)T cells was markedly reduced in the Tg mice. This difference in T cell numbers correlated with a reduced capacity of these cells to proliferate after stimulation and was associated with a major defect in the ability of CD4(+) T cells from infected mice to produce IFN-gamma. Together, these studies reveal that inhibition of NF-kappa B activity in T and NK cells results in defective effector cell expansion and production of IFN-gamma required for resistance to T. gondii. PMID- 12626573 TI - Human NKT cells express granulysin and exhibit antimycobacterial activity. AB - Human NKT cells are a unique subset of T cells that express an invariant V alpha 24 TCR that recognizes the nonclassical Ag-presenting molecule CD1d. Activation of NKT cells is greatly augmented by the marine sponge-derived glycolipid alpha galactosylceramide (alpha GalCer). Because human monocyte-derived cells express CD1d and can harbor the intracellular pathogen Mycobacterium tuberculosis, we asked whether the addition of alpha GalCer could be used to induce effector functions of NKT cells against infected monocytes, macrophages, and monocyte derived dendritic cells. NKT cells secreted IFN-gamma, proliferated, and exerted lytic activity in response to alpha GalCer-pulsed monocyte-derived cells. Importantly, alpha GalCer-activated NKT cells restricted the growth of intracellular M. tuberculosis in a CD1d-dependent manner. NKT cells that exhibited antimycobacterial activity also expressed granulysin, an antimicrobial peptide shown to mediate an antimycobacterial activity through perturbation of the mycobacterial surface. Degranulation of NKT cells resulted in depletion of granulysin and abrogation of antimycobacterial activity. The detection of CD1d in granulomas of tuberculosis patients supports the potential interaction of NKT cells with CD1d-expressing cells at the site of disease activity. These studies provide evidence that alpha Gal Cer-activated CD1d-restricted T cells can participate in human host defense against M. tuberculosis infection. PMID- 12626574 TI - Immunogene therapy of tumors with vaccine based on xenogeneic epidermal growth factor receptor. AB - The breaking of immune tolerance against self epidermal growth factor receptor (EGFr) should be a useful approach for the treatment of receptor-positive tumors with active immunization. To test this concept, we constructed a plasmid DNA encoding extracellular domain of xenogeneic (human) EGFr (hEe-p) or corresponding control mouse EGFr (mEe-p) and empty vector (c-p). Mice immunized with hEe-p showed both protective and therapeutic antitumor activity against EGFr-positive tumor. Sera isolated from the hEe-p-immunized mice exhibited positive staining for EGFr-positive tumor cells in flow cytometric analysis and recognized a single 170-kDa band in Western blot analysis. Ig subclasses responded to rEGFr proteins were elevated in IgG1, Ig2a, and Ig2b. There was the deposition of IgG on the tumor cells. Adoptive transfer of the purified Igs showed the antitumor activity. The increased killing activity of CTL against EGFr-positive tumor cells could be blocked by anti-CD8 or anti-MHC class I mAb. In vivo depletion of CD4(+) T lymphocytes could completely abrogate the antitumor activity, whereas the depletion of CD8(+) cells showed partial abrogation. The adoptive transfer of CD4 depleted (CD8(+)) or CD8-depleted (CD4(+)) T lymphocytes isolated from mice immunized with hEe-p vaccine showed the antitumor activity. In addition, the increase in level of both IFN-gamma and IL-4 was found. Taken together, these findings may provide a new vaccine strategy for the treatment of EGFr-positive tumors through the induction of the autoimmune response against EGFr in a cross reaction between the xenogeneic homologous and self EGFr. PMID- 12626575 TI - Dendritic cell (DC)-based protection against an intracellular pathogen is dependent upon DC-derived IL-12 and can be induced by molecularly defined antigens. AB - Upon loading with microbial Ag and adoptive transfer, dendritic cells (DC) are able to induce immunity to infections. This offers encouragement for the development of DC-based vaccination strategies. However, the mechanisms underlying the adjuvant effect of DC are not fully understood, and there is a need to identify Ag with which to arm DC. In the present study, we analyzed the role of DC-derived IL-12 in the induction of resistance to Leishmania major, and we evaluated the protective efficacy of DC loaded with individual Leishmania Ag. Using Ag-pulsed Langerhans cells (LC) from IL-12-deficient or wild-type mice for immunization of susceptible animals, we showed that the inability to release IL 12 completely abrogated the capacity of LC to mediate protection against leishmaniasis. This suggests that the availability of donor LC-derived IL-12 is a requirement for the development of protective immunity. In addition, we tested the protective effect of LC loaded with Leishmania homolog of receptor for activated C kinase, gp63, promastigote surface Ag, kinetoplastid membrane protein 11, or Leishmania homolog of eukaryotic ribosomal elongation and initiation factor 4a. The results show that mice vaccinated with LC that had been pulsed with selected molecularly defined parasite proteins are capable of controlling infection with L. major. Moreover, the protective potential of DC pulsed with a given Leishmania Ag correlated with the level of their IL-12 expression. Analysis of the cytokine profile of mice after DC-based vaccination revealed that protection was associated with a shift toward a Th1-type response. Together, these findings emphasize the critical role of IL-12 produced by the sensitizing DC and suggest that the development of a DC-based subunit vaccine is feasible. PMID- 12626576 TI - CD40 ligand trimer enhances the response of CD8+ T cells to Mycobacterium tuberculosis. AB - We investigated the effect of recombinant CD40 ligand trimer (CD40LT) on the functional capacity of peripheral blood CD8(+) T cells from healthy tuberculin reactors that were cultured with Mycobacterium tuberculosis-infected autologous monocytes. CD40LT enhanced the capacity of M. tuberculosis-responsive CD8(+) T cells to produce IFN-gamma by increasing the number of IFN-gamma-producing CD8(+) T cells and the amount of IFN-gamma produced per cell. CD40LT-induced IFN-gamma production was dependent on production of IL-12 and IL-18, but did not require IL 15. CD40LT up-regulated expression of the transcription factors phosphorylated CREB and c-Jun, both of which have been previously shown to stimulate IFN-gamma mRNA transcription by binding to the IFN-gamma promoter. CD40LT also enhanced the capacity of CD8(+) T cells to lyse M. tuberculosis-infected monocytes, and increased CTL activity was associated with higher expression of perforin and granulysin, but not of Fas ligand. We conclude that CD40LT can enhance CD8(+) T cell effector function in response to M. tuberculosis. PMID- 12626577 TI - CD4+ T cell-induced differentiation of EBV-transformed lymphoblastoid cells is associated with diminished recognition by EBV-specific CD8+ cytotoxic T cells. AB - EBV transformation of human B cells in vitro results in establishment of immortalized cell lines (lymphoblastoid cell lines (LCL)) that express viral transformation-associated latent genes and exhibit a fixed, lymphoblastoid phenotype. In this report, we show that CD4(+) T cells can modify the differentiation state of EBV-transformed LCL. Coculture of LCL with EBV-specific CD4(+) T cells resulted in an altered phenotype, characterized by elevated CD38 expression and decreased proliferation rate. Relative to control LCL, the cocultured LCL were markedly less susceptible to lysis by EBV-specific CD8(+) CTL. In contrast, CD4(+) T cell-induced differentiation of LCL did not diminish sensitivity of LCL to lysis by CD8(+) CTL specific for an exogenously loaded peptide Ag or lysis by alloreactive CD8(+) CTL, suggesting that differentiation is not associated with intrinsic resistance to CD8(+) T cell cytotoxicity and that evasion of lysis is confined to EBV-specific CTL responses. CD4(+) T cell induced differentiation of LCL and concomitant resistance of LCL to lysis by EBV specific CD8(+) CTL were associated with reduced expression of viral latent genes. Finally, transwell cocultures, in which direct LCL-CD4(+) T cell contact was prevented, indicated a major role for CD4(+) T cell cytokines in the differentiation of LCL. PMID- 12626578 TI - The dendritic cell-specific chemokine, dendritic cell-derived CC chemokine 1, enhances protective cell-mediated immunity to murine malaria. AB - Cell-mediated immunity plays a crucial role in the control of many infectious diseases, necessitating the need for adjuvants that can augment cellular immune responses elicited by vaccines. It is well established that protection against one such disease, malaria, requires strong CD8(+) T cell responses targeted against the liver stages of the causative agent, Plasmodium spp. In this report we show that the dendritic cell-specific chemokine, dendritic cell-derived CC chemokine 1 (DC-CK1), which is produced in humans and acts on naive lymphocytes, can enhance Ag-specific CD8(+) T cell responses when coadministered with either irradiated Plasmodium yoelii sporozoites or a recombinant adenovirus expressing the P. yoelii circumsporozoite protein in mice. We further show that these enhanced T cell responses result in increased protection to malaria in immunized mice challenged with live P. yoelii sporozoites, revealing an adjuvant activity for DC-CK1. DC-CK1 appears to act preferentially on naive mouse lymphocytes, and its adjuvant effect requires IL-12, but not IFN-gamma or CD40. Overall, our results show for the first time an in vivo role for DC-CK1 in the establishment of primary T cell responses and indicate the potential of this chemokine as an adjuvant for vaccines against malaria as well as other diseases in which cellular immune responses are important. PMID- 12626580 TI - Serum-resistant strains of Borrelia burgdorferi evade complement-mediated killing by expressing a CD59-like complement inhibitory molecule. AB - Borrelia burgdorferi, the etiological agent of Lyme disease, comprises three genospecies, Borrelia garinii, afzelii, and burgdorferi sensu strictu, that exhibit different pathogenicity and differ in the susceptibility to C-mediated killing. We examined C-sensitive and C-resistant strains of B. burgdorferi for deposition of C3 and late C components by fluorescence microscope and flow cytometry. Despite comparable deposition of C3 on the two strains, the resistant strain exhibited reduced staining for C6 and C7, barely detectable C9, and undetectable poly C9. Based on these findings, we searched for a protein that inhibits assembly of C membrane attack complex and documented an anti-human CD59 reactive molecule on the surface of C-resistant spirochetes by flow cytometry and electron microscopy. A molecule of 80 kDa recognized by polyclonal and monoclonal anti-CD59 Abs was identified in the membrane extract of C-resistant strains by SDS-PAGE and Western blot analysis. The molecule was released from the bacterial wall using deoxycholate and trypsin, suggesting its insertion into the bacterial membrane. The CD59-like molecule acts as C inhibitor on Borrelia because incubation with F(ab')(2) anti-CD59 renders the serum-resistant strain exquisitely susceptible to C-mediated killing and guinea pig erythrocytes bearing C5b-8, unlike the RBC coated with C5b-7, are protected from reactive lysis by the bacterial extract. Western blot analysis revealed preferential binding of the C inhibitory molecule to C9 and weak interaction with C8 beta. PMID- 12626579 TI - Perforin-mediated effector function within the central nervous system requires IFN-gamma-mediated MHC up-regulation. AB - CD8(+) T cells infiltrating the CNS control infection by the neurotropic JHM strain of mouse hepatitis virus. Differential susceptibility of infected cell types to clearance by perforin or IFN-gamma uncovered distinct, nonredundant roles for these antiviral mechanisms. To separately evaluate each effector function specifically in the context of CD8(+) T cells, pathogenesis was analyzed in mice deficient in both perforin and IFN-gamma (PKO/GKO) or selectively reconstituted for each function by transfer of CD8(+) T cells. Untreated PKO/GKO mice were unable to control the infection and died of lethal encephalomyelitis within 16 days, despite substantially higher CD8(+) T cell accumulation in the CNS compared with controls. Uncontrolled infection was associated with limited MHC class I up-regulation and an absence of class II expression on microglia, coinciding with decreased CD4(+) T cells in CNS infiltrates. CD8(+) T cells from perforin-deficient and wild-type donors reduced virus replication in PKO/GKO recipients. By contrast, IFN-gamma-deficient donor CD8(+) T cells did not affect virus replication. The inability of perforin-mediated mechanisms to control virus in the absence of IFN-gamma coincided with reduced class I expression. These data not only confirm direct antiviral activity of IFN-gamma within the CNS but also demonstrate IFN-gamma-dependent MHC surface expression to guarantee local T cell effector function in tissues inherently low in MHC expression. The data further imply that IFN-gamma plays a crucial role in pathogenesis by regulating the balance between virus replication in oligodendrocytes, CD8(+) T cell effector function, and demyelination. PMID- 12626581 TI - Lupus-prone mice have an abnormal response to thioglycolate and an impaired clearance of apoptotic cells. AB - Deficiency of complement in humans and mice is associated with the development of lupus and with abnormal repair of inflammatory and immune complex-mediated tissue injury. Here we ask whether similar defects in the resolution of inflammation are found in mice prone to spontaneous lupus. We compared the response to an i.p. injection of thioglycolate between two lupus-prone strains (MRL/Mp and NZB/W) and two non lupus-prone strains of mice (C57BL/6 and BALB/c). In all four strains the influx of polymorphonuclear neutrophils (PMN) was similar. However, by 96 h clearance of PMN in the control strains was complete, whereas in the autoimmune prone strains PMN were still detectable. The number of mononuclear cells recruited was markedly reduced in the lupus-prone strains compared with the controls, and their phenotype was different. The lupus-prone strains had significantly fewer elicited macrophages that were CD11b-high and Ly6C-negative. In lupus-prone mice at 24 h there was a significantly increased number of apoptotic PMN free in the peritoneum, accompanied by a reduced percentage of macrophages containing apoptotic bodies, suggesting a defect in their uptake. An impaired ability of resident peritoneal macrophages from lupus-prone mice to engulf apoptotic cells was demonstrated by in vivo and in vitro cell clearance assays. These observations indicate that lupus-prone strains have an abnormal inflammatory response to thioglycolate and an intrinsic impairment in apoptotic cell uptake. These findings have implications for the initiation of autoimmunity, as lupus autoantigens are expressed on dying cells, and impaired disposal of these could enhance the development of autoimmunity. PMID- 12626582 TI - Proinflammatory activities of S100: proteins S100A8, S100A9, and S100A8/A9 induce neutrophil chemotaxis and adhesion. AB - S100A8 and S100A9 are small calcium-binding proteins that are highly expressed in neutrophil and monocyte cytosol and are found at high levels in the extracellular milieu during inflammatory conditions. Although reports have proposed a proinflammatory role for these proteins, their extracellular activity remains controversial. In this study, we report that S100A8, S100A9, and S100A8/A9 caused neutrophil chemotaxis at concentrations of 10(-12)-10(-9) M. S100A8, S100A9, and S100A8/A9 stimulated shedding of L-selectin, up-regulated and activated Mac-1, and induced neutrophil adhesion to fibrinogen in vitro. Neutralization with Ab showed that this adhesion was mediated by Mac-1. Neutrophil adhesion was also associated with an increase in intracellular calcium levels. However, neutrophil activation by S100A8, S100A9, and S100A8/A9 did not induce actin polymerization. Finally, injection of S100A8, S100A9, or S100A8/A9 into a murine air pouch model led to rapid, transient accumulation of neutrophils confirming their activities in vivo. These studies 1) show that S100A8, S100A9, and S100A8/A9 are potent stimulators of neutrophils and 2) strongly suggest that these proteins are involved in neutrophil migration to inflammatory sites. PMID- 12626583 TI - Pre-existing glomerular immune complexes induce polymorphonuclear cell recruitment through an Fc receptor-dependent respiratory burst: potential role in the perpetuation of immune nephritis. AB - In immune complex (IC) diseases, FcR are essential molecules facilitating polymorphonuclear cell (PMN) recruitment and effector functions at the IC site. Although FcR-dependent initial tethering and FcR/integrin-dependent PMN accumulation were postulated, their underlying mechanisms remain unclear. We here addressed potential mechanisms involved in PMN recruitment in acute IC glomerulonephritis (nephrotoxic nephritis). Since some renal cells may be recruited from bone marrow (BM) lineages, reconstitution studies with BM chimeras and PMN transfer between wild-type (WT) and FcR-deficient mice (gamma(-/-)) were performed. Severe glomerular damage was induced in WT and W gamma chimeras (BM from WT to irradiated gamma(-/-)), while it was absent in gamma(-/-) and gamma W chimeras (gamma(-/-) BM to WT). Moreover, WT PMN transfer, but not gamma(-/-) PMN, reconstituted the disease in gamma(-/-), indicating that FcR on resident cells is not a prerequisite for PMN recruitment in this disease. Surprisingly, transferred WT PMN were recruited coincidentally with NF-kappa B activation and TNF-alpha overexpression even in glomeruli with preformed IC (nephrotoxic Ab administered 3 days previously), suggesting that PMN can initially be recruited via its own FcR without previous chemoattractant release. Furthermore, H(2)O(2) inhibition by catalase attenuated the acute WT PMN recruitment and the induction of NF-kappa B and TNF-alpha much more than integrin (CD18) blockade, indicating a role for the respiratory burst before integrin-dependent accumulation. In coculture experiments with IC-stimulated PMN and glomeruli, PMN caused acute glomerular TNF-alpha expression predominantly via FcR-mediated H(2)O(2) production. In conclusion, glomerular IC, even preformed, can cause PMN recruitment and injury through PMN FcR-mediated respiratory burst during initial PMN tethering to IC. PMID- 12626584 TI - Reduced macrophage recruitment, proliferation, and activation in colony stimulating factor-1-deficient mice results in decreased tubular apoptosis during renal inflammation. AB - Kidney tubular epithelial cell (TEC) death may be dependent on the number and activation state of macrophages (M phi) during inflammation. Our prior studies indicate that activated M phi release soluble mediators that incite TEC death, and reducing intrarenal M phi during kidney disease diminishes TEC apoptosis. CSF 1 is required for M phi proliferation and survival. We hypothesized that in the absence of CSF-1, M phi-mediated TEC apoptosis would be prevented during renal inflammation. To test this hypothesis, we evaluated renal inflammation during unilateral ureter obstruction in CSF-1-deficient (Csf1(op)/Csf1(op)) mice. We detected fewer M phi and T cells and less apoptotic TEC in the obstructed kidneys of Csf1(op)/Csf1(op) mice compared with wild-type (WT) mice. The decrease in intrarenal M phi resulted from diminished recruitment and proliferation, not enhanced apoptosis. CSF-1 enhanced M phi activation. There were far fewer activated (CD69, CD23, Ia, surface expression) M phi in obstructed CSF-1 deficient compared with WT obstructed kidneys. Similarly, bone marrow M phi preincubated with anti-CSF-1 receptor Ab or anti-CSF-1 neutralizing Ab were resistant to LPS- and IFN-gamma-induced activation. We detected fewer apoptotic inducing molecules (reactive oxygen species, TNF-alpha, inducible NO synthase) in 1) M phi propagated from obstructed Csf1(op)/Csf1(op) compared with WT kidneys, and 2) WT bone marrow M phi blocked with anti-CSF-1 receptor or anti-CSF-1 Ab compared with the isotype control. Furthermore, blocking CSF-1 or the CSF-1 receptor induced less TEC apoptosis than the isotype control. We suggest that during renal inflammation, CSF-1 mediates M phi recruitment, proliferation, activation, and, in turn, TEC apoptosis. PMID- 12626585 TI - Activation of STAT3 by IL-6 and IL-10 in primary human macrophages is differentially modulated by suppressor of cytokine signaling 3. AB - On human macrophages IL-10 acts as a more potent anti-inflammatory cytokine than IL-6, although both cytokines signal mainly via activation of the transcription factor STAT3. In this study we compare IL-10 and IL-6 signaling in primary human macrophages derived from blood monocytes. Pretreatment of macrophages with PMA or the proinflammatory mediators LPS and TNF-alpha blocks IL-6-induced STAT3 activation, whereas IL-10-induced activation of STAT3 remains largely unaffected. Although LPS induces the feedback inhibitor suppressor of cytokine signaling 3 (SOCS3) in macrophages, inhibition of IL-6 signal transduction by LPS occurs rapidly and does not depend on gene transcription. We also found that pretreatment of macrophages with IL-10 inhibits subsequent STAT3 activation by IL 6, whereas IL-10-induced STAT3 activation is not affected by preincubation with IL-6. This cross-inhibition is dependent on active transcription and might therefore be explained by different sensitivities of IL-10 and IL-6 signaling toward the feedback inhibitor SOCS3, which is induced by both cytokines. In contrast to the IL-6 signal transducer gp130, which has been previously shown to recruit SOCS3 to one of its phosphotyrosine residues (Y759), peptide precipitation experiments suggest that SOCS3 does not interact with phosphorylated tyrosine motifs of the IL-10R. Taken together, different sensitivities of IL-10 and IL-6 signaling toward mechanisms that inhibit the Janus kinase/STAT pathway define an important mechanism that contributes to the different anti-inflammatory potencies of these two cytokines. PMID- 12626587 TI - Secretory phospholipases A2 activate selective functions in human eosinophils. AB - Secretory phospholipases A(2) (sPLA(2)s) are released in large amounts in the blood of patients with systemic inflammatory diseases and accumulate at sites of chronic inflammation, such as the airways of patients with bronchial asthma. Blood eosinophils or eosinophils recruited in inflammatory areas therefore can be exposed in vivo to high concentrations of sPLA(2). We have examined the effects of two structurally different sPLA(2)s (group IA and group IIA) on several functions of eosinophils isolated from normal donors and patients with hypereosinophilia. Both group IA and IIA sPLA(2) induced a concentration dependent release of beta-glucuronidase, IL-6, and IL-8. Release of the two cytokines was associated with the accumulation of their specific mRNA. In addition, sPLA(2)s induced the surface expression of CD44 and CD69, two major activation markers of eosinophils. In contrast, none of the sPLA(2)s examined induced the production of IL-5, the de novo synthesis of leukotriene C(4) and platelet-activating factor, or the generation of superoxide anion from human eosinophils. Incubation of eosinophils with the major enzymatic products of the sPLA(2)s (arachidonic acid, lysophosphatidylcholine, or lysophosphatidic acid) did not reproduce any of the enzymes' effects. In addition, inactivation of sPLA(2) enzymatic activity by bromophenacyl bromide did not influence the release of beta-glucuronidase or of cytokines. Stimulation of eosinophils by sPLA(2)s was associated with activation of extracellular signal-regulated kinases 1/2. These results indicate that sPLA(2)s selectively activate certain proinflammatory and immunoregulatory functions of human eosinophils through mechanism(s) independent from enzymatic activity and from the generation of arachidonic acid. PMID- 12626586 TI - Monocytes are potent facilitators of alveolar neutrophil emigration during lung inflammation: role of the CCL2-CCR2 axis. AB - Coordinated neutrophil and monocyte recruitment is a characteristic feature of acute lung inflammatory responses. We investigated the role of monocyte chemotactic protein-1 (CCL2, JE) and the chemokine receptor CCR2 in regulating alveolar leukocyte traffic. Groups of wild-type (WT) mice, CCR2-deficient mice, lethally irradiated CCR2-deficient and WT mice that were reciprocally bone marrow transplanted (chimeric CCR2 deficient and WT, respectively), chimeric CCR2 deficient mice with an enriched CCR2(+) alveolar macrophage population, and CCR2 deficient mice transfused with CCR2(+) mononuclear cells were treated with intratracheal CCL2 and/or Escherichia coli endotoxin. Our data show that alveolar monocyte recruitment is strictly dependent on CCR2. LPS-induced neutrophil migration to the lungs is CCR2 independent. However, when CCR2-bearing blood monocytes are present, alveolar neutrophil accumulation is accelerated and drastically amplified. We suggest that this hitherto unrecognized cooperativity between monocytes and neutrophils contributes to the strong, coordinated leukocyte efflux in lung inflammation. PMID- 12626588 TI - Engineered recombinant peanut protein and heat-killed Listeria monocytogenes coadministration protects against peanut-induced anaphylaxis in a murine model. AB - Peanut allergy (PNA) is the major cause of fatal and near-fatal anaphylactic reactions to foods. Traditional immunotherapy using peanut (PN) protein is not an option for PNA therapy because of the high incidence of adverse reactions. We investigated the effects of s.c. injections of engineered (modified) recombinant PN proteins and heat-killed Listeria monocytogenes (HKLM) as an adjuvant on anaphylactic reactions in a mouse model of PN allergy. PN-allergic C3H/HeJ mice were treated s.c. with a mixture of the three major PN allergens and HKLM (modified (m)Ara h 1-3 plus HKLM). The effects on anaphylactic reactions following PN challenge and the association with Ab levels and cytokine profiles were determined. Although all mice in the sham-treated groups exhibited anaphylactic symptoms with a median symptom score of 3, only 31% of mice in the mAra h 1-3 plus HKLM group developed mild anaphylaxis, with a low median symptom score of 0.5. Alterations in core body temperature, bronchial constriction, plasma histamine, and PN-specific IgE levels were all significantly reduced. This protective effect was markedly more potent than in the mAra h 1-3 protein alone treated group. HKLM alone did not have any protective effect. Reduced IL-5 and IL 13, and increased IFN-gamma levels were observed only in splenocytes cultures from mAra h 1-3 plus HKLM-treated mice. These results show that immunotherapy with modified PN proteins and HKLM is effective for treating PN allergy in this model, and may be a potential approach for treating PNA. PMID- 12626589 TI - A causative relationship exists between eosinophils and the development of allergic pulmonary pathologies in the mouse. AB - Asthma and mouse models of allergic respiratory inflammation are invariably associated with a pulmonary eosinophilia; however, this association has remained correlative. In this report, a causative relationship between eosinophils and allergen-provoked pathologies was established using eosinophil adoptive transfer. Eosinophils were transferred directly into the lungs of either naive or OVA treated IL-5(-/-) mice. This strategy resulted in a pulmonary eosinophilia equivalent to that observed in OVA-treated wild-type animals. A concomitant consequence of this eosinophil transfer was an increase in Th2 bronchoalveolar lavage cytokine levels and the restoration of intracellular epithelial mucus in OVA-treated IL-5(-/-) mice equivalent to OVA-treated wild-type levels. Moreover, the transfer also resulted in the development of airway hyperresponsiveness. These pulmonary changes did not occur when eosinophils were transferred into naive IL-5(-/-) mice, eliminating nonspecific consequences of the eosinophil transfer as a possible explanation. Significantly, administration of OVA-treated IL-5(-/-) mice with GK1.5 (anti-CD4) Abs abolished the increases in mucus accumulation and airway hyperresponsiveness following adoptive transfer of eosinophils. Thus, CD4(+) T cell-mediated inflammatory signals as well as signals derived from eosinophils are each necessary, yet alone insufficient, for the development of allergic pulmonary pathology. These data support an expanded view of T cell and eosinophil activities and suggest that eosinophil effector functions impinge directly on lung function. PMID- 12626590 TI - IL-4 down-regulates anaphylatoxin receptors in monocytes and dendritic cells and impairs anaphylatoxin-induced migration in vivo. AB - Anaphylatoxins mobilize leukocytes to the sites of inflammation. In the present study we investigated the impact of GM-CSF, IL-4, and IFN-gamma on anaphylatoxin receptor expression in monocytes and dendritic cells (DC). IL-4 was identified as the strongest down-regulator of the receptors for C5a and C3a in monocytes and monocyte-derived DC (MoDC). To study the impact of IL-4 on anaphylatoxin-induced chemotaxis, an in vivo migration model was established. For this purpose, human monocytes and MoDC were injected i.v. into SCID mice that at the same time received anaphylatoxins into the peritoneal cavity. A peritoneal influx of human monocytes could be demonstrated by 4 h after injections of C5a and C3a. In line with receptor down-regulation, IL-4 treatment inhibited in vivo mobilization of human monocytes and MoDC in response to C5a and C3a. In addition to its effects on human cells, IL-4 reduced C5a receptors in murine bone marrow-derived DC and impaired recruitment of labeled bone marrow-derived DC in syngeneic BALB/c mice to i.p. injected C5a. Overall, these data suggest that inhibition of a rapid anaphylatoxin-induced mobilization of monocytes and DC to inflamed tissues represents an important anti-inflammatory activity of the Th2 cytokine IL-4. PMID- 12626591 TI - IFN-gamma reverses the stop signal allowing migration of antigen-specific T cells into inflammatory sites. AB - In humans the majority of endothelial cells (EC) constitutively express MHC class II Ags. We know that in vitro ECs can activate CD45RO(+) B7-independent CD4(+) T cells to proliferate and produce IL-2. The in vivo correlate of this T cell response is not known, and here we have explored whether endothelial expression of MHC class II Ags affects the transendothelial migration of alloreactive CD4(+) CD45RO(+) B7-independent T cells. Alloreactive CD4(+) T cell clones and lines were generated against HLA-DR11, DR13, DR4, and DR1 MHC Ags, and their rates of migration across untreated EC line Eahy.926 (MHC class II negative) or Eahy.926 transfected with CIITA (EahyCIITA) to express DR11 and DR13 were investigated. The migrations of EahyCIITA-specific T cell clones and lines were retarded in a DR-specific manner, and retardation was reversed in the presence of mAb to DR Ag. When investigating the ability of T cells to proliferate in response to EahyCIITA before and after transmigration, migrated cells were still able to proliferate, but the frequency of EahyCIITA-specific cells was much reduced compared with that of nonmigrated cells. The use of fluorescently labeled T cells revealed that specific cells become trapped within the endothelial monolayer. Pretreatment of EahyCIITA with IFN-gamma restored the ability of DR11- or DR13-specific T cells to transmigrate and proliferate, thus abrogating DR-specific retardation. We conclude that cognate interaction between T cells and endothelial MHC class II initiates a stop signal possibly similar to an immunological synapse, but this is overcome in an inflammatory milieu. PMID- 12626593 TI - Neutrophils promote mononuclear cell infiltration during viral-induced encephalitis. AB - Neutrophils are the first infiltrating cell population to appear within the CNS during infection with the neurotropic JHM strain of mouse hepatitis virus (JHMV). To determine whether neutrophils play a role in limiting acute JHMV infection, mice were depleted of neutrophils. Infection of neutropenic animals resulted in increased levels of virus replication and mortality compared with control mice. Furthermore, neutropenia resulted in significantly reduced mononuclear leukocyte infiltration possibly due to reduced loss of blood brain barrier integrity during acute JHMV infection. These data suggest that infiltrating neutrophils are crucial for limiting virus replication during acute JHMV infection, contribute to the loss of blood brain barrier integrity and play a role in shaping adaptive immunity within the CNS. PMID- 12626592 TI - Redundancy of a functional melanocortin 1 receptor in the anti-inflammatory actions of melanocortin peptides: studies in the recessive yellow (e/e) mouse suggest an important role for melanocortin 3 receptor. AB - The issue of which melanocortin receptor (MC-R) is responsible for the anti inflammatory effects of melanocortin peptides is still a matter of debate. Here we have addressed this aspect using a dual pharmacological and genetic approach, taking advantage of the recent characterization of more selective agonists/antagonists at MC1 and MC3-R as well as of the existence of a naturally defective MC1-R mouse strain, the recessive yellow (e/e) mouse. RT-PCR and ultrastructural analyses showed the presence of MC3-R mRNA and protein in peritoneal macrophages (M phi) collected from recessive yellow (e/e) mice and wild-type mice. This receptor was functional as Mphi incubation (30 min) with melanocortin peptides led to accumulation of cAMP, an effect abrogated by the MC3/4-R antagonist SHU9119, but not by the selective MC4-R antagonist HS024. In vitro M phi activation, determined as release of the CXC chemokine KC and IL-1 beta, was inhibited by the more selective MC3-R agonist gamma(2)-melanocyte stimulating hormone but not by the selective MC1-R agonist MS05. Systemic treatment of mice with a panel of melanocortin peptides inhibited IL-1 beta release and PMN accumulation elicited by urate crystals in the murine peritoneal cavity. MS05 failed to inhibit any of the inflammatory parameters either in wild type or recessive yellow (e/e) mice. SHU9119 prevented the inhibitory actions of gamma(2)-melanocyte stimulating hormone both in vitro and in vivo while HS024 was inactive in vivo. In conclusion, agonism at MC3-R expressed on peritoneal M phi leads to inhibition of experimental nonimmune peritonitis in both wild-type and recessive yellow (e/e) mice. PMID- 12626594 TI - The cytokine macrophage migration inhibitory factor reduces pro-oxidative stress induced apoptosis. AB - The cytokine macrophage migration inhibitory factor (MIF) exhibits pro- and anti inflammatory activities and regulates cell proliferation and survival. We investigated the effects of MIF on apoptosis. As MIF exhibits oxidoreductase activity and participates in regulating oxidative cell stress, we studied whether MIF could affect oxidative stress-induced apoptosis. We demonstrated that MIF exhibits antiapoptotic activity in various settings. MIF suppressed camptothecin induced apoptosis in HeLa and Kym cells and HL-60 promyeloblasts. Both exogenous MIF and endogenous MIF, induced following overexpression through tetracycline (tet) gene induction, led to significant suppression of apoptosis. Apoptosis reduction by MIF was also observed in T cells. A role for MIF in redox stress induced apoptosis was addressed by comparing the effects of rMIF with those of the oxidoreductase mutant C60SMIF. Endogenous overexpression of C60SMIF was similar to that of MIF, but C60SMIF did not suppress apoptosis. Exogenous rC60SMIF inhibited apoptosis. A role for MIF in oxidative stress-induced apoptosis was directly studied in HL-60 leukocytes and tet-regulated HeLa cells following thiol starvation or diamide treatment. MIF protected these cells from redox stress-induced apoptosis and enhanced cellular glutathione levels. As overexpressed C60SMIF did not protect tet-regulated HeLa cells from thiol starvation-induced apoptosis, it seems that the redox motif of MIF is important for this function. Finally, overexpression of MIF inhibited phosphorylation of endogenous c-Jun induced by thiol starvation, indicating that MIF-based suppression of apoptosis is mediated through modulation of c-Jun N-terminal kinase activity. Our findings show that MIF has potent antiapoptotic activities and suggest that MIF is a modulator of pro-oxidative stress-induced apoptosis. PMID- 12626595 TI - CXCR2 regulates respiratory syncytial virus-induced airway hyperreactivity and mucus overproduction. AB - Severe inflammation and mucus overproduction are partially responsible for respiratory syncytial virus (RSV)-induced disease in infants. Using a murine model, we characterized the virally induced chemokine receptors responsible for mediating the pathophysiological response to RSV infection, we found that CXCR2 mRNA was induced at 4 days after RSV infection. Immunohistochemical staining demonstrated that CXCR2 protein was expressed on alveolar macrophages. Immunoneutralization of CXCR2 resulted in decreased airway hyperreactivity relative to the RSV-infected controls. In addition, there was decreased mucus in the bronchoalveolar lavage fluid, decreased periodic-acid Schiff staining, and significantly less mucus-associated gob-5 mRNA and protein in anti-CXCR2-treated mice. The effects of anti-CXCR2 treatment were not a result of differences in viral clearance or neutrophil influx, as these parameters were comparable in both groups of animals. To confirm our immunoneutralization studies, we performed experiments in CXCR2(-/-) mice. Results in CXCR2(-/-) mice recapitulated results from our immunoneutralization studies. However, CXCR2(-/-) mice also showed a statistically significant decrease in muc5ac, relative to RSV-infected wild-type animals. Thus, CXCR2 may be a relevant target in the pathogenesis of RSV bronchiolitis, since it contributes to mucus production and airway hyperreactivity in our model of RSV infection. PMID- 12626597 TI - IL-8 directly enhanced endothelial cell survival, proliferation, and matrix metalloproteinases production and regulated angiogenesis. AB - IL-8, a member of the chemokine family, has been shown to play an important role in tumor growth, angiogenesis, and metastasis. The objective of this study was to determine the mechanism of IL-8-mediated angiogenesis. We examined the direct role of IL-8 in angiogenesis by examining IL-8 receptor expression on endothelial cells and their proliferation, survival, and matrix metalloproteinases (MMPs) production. We demonstrate that HUVEC and human dermal microvascular endothelial cells constitutively express CXCR1 and CXCR2 mRNA and protein. Recombinant human IL-8 induced endothelial cell proliferation and capillary tube organization while neutralization of IL-8 by anti-IL-8 Ab blocks IL-8-mediated capillary tube organization. Incubation of endothelial cells with IL-8 inhibited endothelial cell apoptosis and enhanced antiapoptotic gene expression. Endothelial cells incubated with IL-8 had higher levels of Bcl-x(L):Bcl-x(S) and Bcl-2:Bax ratios. Furthermore, incubation of endothelial cells with IL-8 up-regulated MMP-2 and MMP 9 production and mRNA expression. Our data suggest that IL-8 directly enhanced endothelial cell proliferation, survival, and MMP expression in CXCR1- and CXCR2 expressing endothelial cells and regulated angiogenesis. PMID- 12626596 TI - Down-regulation of proinflammatory capacity during apoptosis in human polymorphonuclear leukocytes. AB - Polymorphonuclear leukocytes (PMNs) are essential to innate immunity in humans and contribute significantly to inflammation. Although progress has been made, the molecular basis for termination of inflammation in humans is incompletely characterized. We used human oligonucleotide microarrays to identify genes encoding inflammatory mediators that were differentially regulated during the induction of apoptosis. One hundred thirty-three of 212 differentially expressed genes encoding proinflammatory factors, signal transduction mediators, adhesion molecules, and other proteins that facilitate the inflammatory response were down regulated during the induction of apoptosis following PMN phagocytosis. Among these, 42 genes encoded proteins critical to the inflammatory response, including receptors for IL-8 beta, IL-10 alpha, IL-13 alpha 1, IL-15 alpha, IL-17, IL-18, C1q, low-density lipoprotein, IgG Fc (CD32), and formyl peptide, Toll-like receptor 6, platelet/endothelial cell adhesion molecule-1 (CD31), P-selectin (CD62), IL-1 alpha, IL-16, and granulocyte chemoattractant protein-2 were down regulated. Many of these genes were similarly down-regulated during Fas-mediated or camptothecin-induced apoptosis. We used flow cytometry to confirm that IL-8R beta (CXCR2) and IL-1 alpha were significantly down-regulated during PMN apoptosis. We also discovered that 23 genes encoding phosphoinositide and calcium mediated signal transduction components, which comprise complex pathways essential to the inflammatory response of host cells, were differentially regulated during PMN apoptosis. Importantly, our data demonstrate that PMNs down regulate proinflammatory capacity at the level of gene expression during induction of apoptosis. These findings provide new insight into the molecular events that resolve inflammation following PMN activation in humans. PMID- 12626598 TI - Macrophage metalloelastase as a major factor for glomerular injury in anti glomerular basement membrane nephritis. AB - Rat anti-glomerular basement membrane (GBM) nephritis is a model of crescentic glomerulonephritis induced by injection of anti-GBM antiserum. To elucidate the mechanism of glomerular injury, we analyzed the gene expression patterns in the kidneys of anti-GBM nephritis rats using DNA arrays, and found that macrophage metalloelastase/matrix metalloproteinase (MMP)-12 was one of the highly expressed genes in the kidneys on days 3 and 7 after the injection of anti-GBM antiserum. Enhancement of MMP-12 mRNA expression was confirmed by Northern blot analysis, and in situ hybridization revealed that MMP-12 mRNA was expressed in ED-1 positive macrophages and multinuclear giant cells in the glomeruli with crescent. Moreover, these cells were positive with anti-rat rMMP-12 Ab on the section of the kidneys of anti-GBM nephritis rats on day 7. To clarify the role of MMP-12, we conducted a neutralization experiment using anti-rat rMMP-12 Ab, which had an ability to inhibit rMMP-12 activity of degrading natural substrate such as bovine elastin or human fibronectin in vitro. Anti-rat rMMP-12 Ab or control Ig was injected in each of six rats on days 0, 2, 4, and 6 after the injection of anti GBM antiserum. Consequently, crescent formation and macrophage infiltration in the glomeruli were significantly reduced in the rats treated with anti-rat rMMP 12 Ab, and the amount of urine protein was also decreased. These results disclosed that MMP-12 played an important role in glomerular injury in a crescentic glomerulonephritis model, and inhibition of MMP-12 may lead to a new therapeutic strategy for this disease. PMID- 12626599 TI - A broad-spectrum caspase inhibitor attenuates allergic airway inflammation in murine asthma model. AB - Asthma is characterized by acute and chronic airway inflammation, and the severity of the airway hyperreactivity correlates with the degree of inflammation. Many of the features of lung inflammation observed in human asthma are reproduced in OVA-sensitized/challenged mice. T lymphocytes, particularly Th2 cells, are critically involved in the genesis of the allergic response to inhaled Ag. In addition to antiapoptotic effects, broad-spectrum caspase inhibitors inhibit T cell activation in vitro. We investigated the effect of the broad spectrum caspase inhibitor, N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (z VAD-fmk), on airway inflammation in OVA-sensitized/challenged mice. OVA sensitized mice treated with z-VAD-fmk immediately before allergen challenge showed marked reduction in inflammatory cell infiltration in the airways and pulmonary blood vessels, mucus production, and Th2 cytokine production. We hypothesized that the caspase inhibitor prevented T cell activation, resulting in the reduction of cytokine production and eosinophil infiltration. Treatment with z-VAD-fmk in vivo prevented subsequent T cell activation ex vivo. We propose that caspase inhibitors may offer a novel therapeutic approach to T cell-dependent inflammatory airway diseases. PMID- 12626600 TI - Role of the chemokine stromal cell-derived factor 1 in autoantibody production and nephritis in murine lupus. AB - In normal mice, stromal cell-derived factor 1 (SDF-1/CXCL12) promotes the migration, proliferation, and survival of peritoneal B1a (PerB1a) lymphocytes. Because these cells express a self-reactive repertoire and are expanded in New Zealand Black/New Zealand White (NZB/W) mice, we tested their response to SDF-1 in such mice. PerB1a lymphocytes from NZB/W mice were exceedingly sensitive to SDF-1. This greater sensitivity was due to the NZB genetic background, it was not observed for other B lymphocyte subpopulations, and it was modulated by IL-10. SDF-1 was produced constitutively in the peritoneal cavity and in the spleen. It was also produced by podocytes in the glomeruli of NZB/W mice with nephritis. The administration of antagonists of either SDF-1 or IL-10 early in life prevented the development of autoantibodies, nephritis, and death in NZB/W mice. Initiation of anti-SDF-1 mAb treatment later in life, in mice with established nephritis, inhibited autoantibody production, abolished proteinuria and Ig deposition, and reversed morphological changes in the kidneys. This treatment also counteracted B1a lymphocyte expansion and T lymphocyte activation. Therefore, PerB1a lymphocytes are abnormally sensitive to the combined action of SDF-1 and IL-10 in NZB/W mice, and SDF-1 is key in the development of autoimmunity in this murine model of lupus. PMID- 12626601 TI - CTLA-4 blockade enhances the therapeutic effect of an attenuated poxvirus vaccine targeting p53 in an established murine tumor model. AB - p53 is overexpressed by half of all cancers, and is an attractive target for a vaccine approach to immunotherapy. p53 overexpression is frequently the result of point mutations, which leaves the majority of the protein in its wild-type form. Therefore, the majority of p53 sequence is wild type, making it a self-protein for which tolerance plays a role in limiting immune responses. To overcome tolerance to p53, we have expressed wild-type murine p53 in the nonpathogenic attenuated poxvirus, modified vaccinia virus Ankara (recombinant modified vaccinia virus Ankara expressing wild-type murine p53 (rMVAp53)). Mice immunized with rMVAp53 vaccine developed vigorous p53-specific CTL responses. rMVAp53 vaccine was evaluated for its ability to inhibit the outgrowth of the syngeneic murine sarcoma Meth A, which overexpresses mutant p53. Mice were inoculated with a lethal dose (5 x 10(5) cells injected s.c.) of Meth A tumor cells and vaccinated by i.p. injection 3 days later with 5 x 10(7) PFU of rMVAp53. The majority of mice remained tumor free and resistant to rechallenge with Meth A tumor cells. We wished to determine whether rMVAp53 immunization could effect the rejection of an established, palpable Meth A tumor. In subsequent experiments, mice were injected with 10(6) Meth A tumor cells, and treated 6 days later with anti-CTLA-4 Ab (9H10) and rMVAp53. The majority of treated mice had complete tumor regression along with lasting tumor immunity. In vivo Ab depletion confirmed that the antitumor effect was primarily CD8 and to a lesser extent CD4 dependent. These experiments demonstrate the potential of a novel cell-free vaccine targeting p53 in malignancy. PMID- 12626602 TI - Prothrombin binds to the surface of apoptotic, but not viable, cells and serves as a target of lupus anticoagulant autoantibodies. AB - Anti-phospholipid Ab (aPL) are a heterogeneous group of autoantibodies directed against various combinations of phospholipids (PL) and PL-binding proteins. Lupus anticoagulant (LA) Ab, a subset of aPL, exhibit anticoagulant properties in vitro, but are procoagulant in vivo. Most LA Ab are specific for either beta(2) glycoprotein I (beta(2)GPI) or prothrombin (PT), two PL-binding proteins. We have previously shown that beta(2)GPI and beta(2)GPI-dependent aPL bind specifically to apoptotic, but not viable, thymocytes. In this study, we demonstrate that PT, like beta(2)GPI, binds selectively to the surface of apoptotic, but not viable, Jurkat cells. Furthermore, PT supports the binding of systemic lupus erythematosus-derived polyclonal and murine monoclonal LA Ab to apoptotic cells. Two LA mAb, which differed dramatically in their relative affinities for PT, were studied. Although one mAb (29J3-62) had a high affinity for PT alone, the other (29I4-24) showed minimal reactivity with PT alone and required PL for elevated binding. Monovalent fragments of 29I4-24 reacted with PL-bound PT with high affinity, suggesting that this mAb recognizes a PL-dependent epitope. Despite these differences, PT-dependent binding of both mAb to apoptotic cells was 30 fold greater than that to viable cells. Moreover, binding of PT to apoptotic cells was, itself, increased in the presence of bivalent, but not monovalent, forms of either mAb. In summary, our data demonstrate the following: 1) specific binding of PT to apoptotic cells, an effect enhanced by PT-dependent LA Ab; 2) heterogeneity of PT-dependent LA Ab; and 3) potential pathogenicity of Ab of either low or high affinity for PT. PMID- 12626603 TI - Cytokine polymorphisms play a role in susceptibility to ultraviolet B-induced modulation of immune responses after hepatitis B vaccination. AB - UVB exposure can alter immune responses in experimental animals and humans. In an earlier human volunteer study, we demonstrated that hepatitis B-specific humoral and cellular immunity after vaccination on average were not significantly affected by UVB exposure. However, it is known that individuals differ in their susceptibility to UVB-induced immunomodulation, and it was hypothesized that polymorphisms in specific cytokines may play a role in this susceptibility. In this respect, we previously demonstrated that immune responses after hepatitis B vaccination are influenced by the minor allelic variant of IL-1 beta in the general population. For all volunteers, single nucleotide polymorphisms were determined for the following UV response-related cytokines: IL-1 receptor antagonist (+2018), IL-1 alpha (+4845), IL-1 beta (+3953), TNF-alpha (-308), and TNF-alpha (-238). Exposure to UVB significantly suppressed Ab responses to hepatitis B in individuals with the minor variant for the IL-1 beta polymorphism. Increased minimal erythema dose values (just perceptible), which resulted in higher absolute UVB exposures, were observed in the same individuals. There were no associations observed between UVB-induced immunomodulation and the other cytokine polymorphisms examined. This study indicates that individual susceptibility to UVB radiation needs to be considered when studying the effects of UVB in humans. PMID- 12626604 TI - Accumulation of B lymphocytes with a naive, resting phenotype in a subset of hepatitis C patients. AB - Chronic infection with hepatitis C virus (HCV) is associated with disturbances of B lymphocyte activation and function: autoantibody production, mixed cryoglobulinemia, and B cell lymphomas. It has been proposed that these abnormalities reflect chronic antigenic stimulation or aberrant signaling through the B cell coreceptor, the latter mediated by binding of the HCV E2 glycoprotein to CD81. To test this hypothesis, we measured expression of activation and differentiation markers on peripheral blood B cells from patients with chronic HCV infection. Thirty-six HCV patients with and without mixed cryoglobulinemia were compared with 18 healthy control volunteers and 17 sustained virologic responders who had cleared HCV infection. Ten of the 36 HCV patient samples showed increased B cell frequencies; B cell frequency was higher in patients with more severe hepatic fibrosis. However, these samples lacked evidence of Ag-driven activation or proliferation. The expanded cells were low in the activation markers CD25, CD69, CD71, CD80, and CD86. Proliferation of circulating B cells was unchanged in HCV patients. These cells did not express the differentiation marker CD27, suggesting that they were not enriched in memory B cells. Furthermore, the expanded B cells expressed both IgD and IgM, suggesting that they were antigenically naive. Together, these results indicate that B cell expansion in the peripheral blood of HCV patients is not associated with Ag mediated activation and differentiation. Instead, factors other than antigenic stimulation may promote the accumulation of peripheral blood B cells with a naive phenotype in a subset of HCV patients. PMID- 12626606 TI - Ontogeny of rhythmic motor patterns generated in the embryonic rat spinal cord. AB - Patterned spontaneous activity is generated in developing neuronal circuits throughout the CNS including the spinal cord. This activity is thought to be important for activity-dependent neuronal growth, synapse formation, and the establishment of neuronal networks. In this study, we examine the spatiotemporal distribution of motor patterns generated by rat spinal cord and medullary circuits from the time of initial axon outgrowth through to the inception of organized respiratory and locomotor rhythmogenesis during late gestation. This includes an analysis of the neuropharmacological control of spontaneous rhythms generated within the spinal cord at different developmental stages. In vitro spinal cord and medullary-spinal cord preparations isolated from rats at embryonic ages (E)13.5-E21.5 were studied. We found age-dependent changes in the spatiotemporal pattern, neurotransmitter control, and propensity for the generation of spontaneous rhythmic motor discharge during the prenatal period. The developmental profile of the neuropharmacological control of rhythmic bursting can be divided into three periods. At E13.5-E15.5, the spinal networks comprising cholinergic and glycinergic synaptic interconnections are capable of generating rhythmic activity, while GABAergic synapses play a role in supporting the spontaneous activity. At late stages (E18.5-E21.5), glutamate drive acting via non- N-methyl-d-aspartate (non-NMDA) receptors is primarily responsible for the rhythmic activity. During the middle stage (E16.5-E17.5), the spontaneous activity results from the combination of synaptic drive acting via non-NMDA glutamatergic, nicotinic acetylcholine, glycine, and GABA(A) receptors. The modulatory actions of chloride-mediated conductances shifts from predominantly excitatory to inhibitory late in gestation. PMID- 12626608 TI - EEG slow (approximately 1 Hz) waves are associated with nonstationarity of thalamo-cortical sensory processing in the sleeping human. AB - Intracellular studies reveal that, during slow wave sleep (SWS), the entire cortical network can swing rhythmically between extremely different microstates, ranging from wakefulness-like network activation to functional disconnection in the space of a few hundred milliseconds. This alternation of states also involves the thalamic neurons and is reflected in the EEG by a slow (<1 Hz) oscillation. These rhythmic changes, occurring in the thalamo-cortical circuits during SWS, may have relevant, phasic effects on the transmission and processing of sensory information. However, brain reactivity to sensory stimuli, during SWS, has traditionally been studied by means of sequential averaging, a procedure that necessarily masks any short-term fluctuation of responsiveness. The aim of this study was to provide a dynamic evaluation of brain reactivity to sensory stimuli in naturally sleeping humans. To this aim, single-trial somatosensory evoked potentials (SEPs) were grouped and averaged as a function of the phase of the ongoing sleep slow (<1 Hz) oscillation. This procedure revealed a dynamic profile of responsiveness, which was conditioned by the phase of the spontaneous sleep EEG. Overall, the amplitude of the evoked potential changed sistematically, increasing and approaching wakefulness levels along the negative slope of the EEG oscillation and decaying below SWS average levels along the positive drift. These marked and fast changes of stimulus-correlated electrical activity involved both short (N20) and long latency (P60 and P100) components of SEPs. In addition, the observed short-term response variability appeared to be centrally generated and specifically related to the evolution of the spontaneous oscillatory pattern. The present findings demonstrate that thalamo-cortical processing of sensory information is not stationary in the very short period (approximately 500 ms) during natural SWS. PMID- 12626607 TI - Effect of extracellular calcium on excitability of guinea pig airway vagal afferent nerves. AB - The effect of reducing extracellular calcium concentration ([Ca(2+)](o)) on vagal afferent excitability was analyzed in a guinea pig isolated vagally innervated trachea-bronchus preparation. Afferent fibers were characterized as either having low-threshold, rapidly adapting mechanosensors (Adelta fibers) or nociceptive like phenotypes (Adelta and C fibers). The nociceptors were derived from neurons within the jugular ganglia, whereas the low-threshold mechanosensors were derived from neurons within the nodose ganglia. Reducing [Ca(2+)](o) did not affect the excitability of the low-threshold mechanosensors in the airway. By contrast, reducing [Ca(2+)](o) selectively increased the excitability of airway nociceptors as manifested by a substantive increase in action potential discharge in response to mechanical stimulation, and in a subset of fibers, by overtly evoking action potential discharge. This increase in the excitability of nociceptors was not mimicked by a combination of omega-conotoxin and nifedipine or tetraethylammonium. Whole cell patch recordings from airway-labeled and unlabeled neurons in the vagal jugular ganglia support the hypothesis that [Ca(2+)](o) inhibits a nonselective cation conductance in vagal nociceptors that may serve to regulate excitability of the nerve terminals within the airways. PMID- 12626609 TI - Adenosine A1 receptor-mediated presynaptic inhibition of GABAergic transmission in immature rat hippocampal CA1 neurons. AB - In the mechanically dissociated rat hippocampal CA1 neurons with native presynaptic nerve endings, namely "synaptic bouton" preparation, the purinergic modulation of spontaneous GABAergic miniature inhibitory postsynaptic currents (mIPSCs) was investigated using whole-cell recording mode under the voltage-clamp conditions. In immature neurons, adenosine (10 microM) reversibly decreased GABAergic mIPSC frequency without affecting the mean current amplitude. The inhibitory effect of adenosine transmission was completely blocked by 8 cyclopentyl-1,3-dipropylxanthine (DPCPX, 100 nM), a selective Alpha(1) receptor antagonist, and was mimicked by N(6)-cyclopentyladenosine (CPA, 1 microM), a selective Alpha(1) receptor agonist. However, CPA had no effect on GABAergic mIPSC frequency in postnatal 30 day neurons. N-ethylmaleimide (10 microM), a guanosine 5'-triphosphate binding protein uncoupler, and Ca(2+)-free external solution removed the CPA-induced inhibition of mIPSC frequency. K(+) channel blockers, 4-aminopyridine (100 microM) and Ba(2+) (1 mM), had no effect on the inhibitory effect of CPA on GABAergic mIPSC frequency. Stimulation of adenylyl cyclase with forskolin (10 microM) prevented the CPA action on GABAergic mIPSC frequency. Rp-cAMPS (100 microM), a selective PKA inhibitor, also blocked the CPA action. It was concluded that the activation of presynaptic Alpha(1) receptors modulates the probability of spontaneous GABA release via cAMP- and protein kinase A dependent pathway. This Alpha(1) receptor-mediated modulation of GABAergic transmission may play an important role in the regulation of excitability of immature hippocampal CA1 neurons. PMID- 12626610 TI - Interjoint coordination in the stick insect leg-control system: the role of positional signaling. AB - Interjoint coordination is essential for proper walking behavior in multi-jointed insect legs. We have shown previously that movement signals from the femur-tibia (FT) joint can shape motor activity of the adjacent coxa-trochanter (CT) joint in the stick insect, Carausius morosus. Here, we present data on the role of position signals from the FT-joint on activity generated in motoneurons (MNs) of the CT-joint. We show that the probability of occurrence of stance (with depression in the CT-joint) or swing movements (with levation in the CT-joint) at the start of walking sequences is influenced by the angle of the FT-joint in the resting animal. We tested the influence of FT-joint angle on pharmacologically induced rhythmic activity of CT-joint depressor (DprTr) and levator (LevTr) MNs. The burst duration, mean spike rate within bursts, and duty cycle for each MN pool were found to depend on FT position. For LevTr MNs, these parameters progressively increased as the FT-joint was moved from extension to flexion, and the opposite was true for DprTr MNs. The cycle period of CT-MN rhythmicity also depended on FT position. In addition, we sometimes observed that the motor output shifted completely to one MN pool at extreme positions, suggesting that the central rhythm-generating network for the CT-joint became locked in one phase. These results indicate that position signals from the FT-joint modulate rhythmic activity in CT-joint MNs partly by having access to central rhythm generating networks of the CT-joint. PMID- 12626611 TI - Heterogeneous actions of serotonin on interneurons in rat visual cortex. AB - The effects of serotonin (5-HT) on excitability of two cortical interneuronal subtypes, fast-spiking (FS) and low threshold spike (LTS) cells, and on spontaneous inhibitory postsynaptic currents (sIPSCs) in layer V pyramidal cells were studied in rat visual cortical slices using whole-cell recording techniques. Twenty-two of 28 FS and 26 of 35 LTS interneurons responded to local application of 5-HT. In the group of responsive neurons, 5-HT elicited an inward current in 50% of FS cells and 15% of LTS cells, an outward current was evoked in 41% of FS cells and 81% of LTS cells, and an inward current followed by an outward current in 9% of FS cells and 4% LTS cells. The inward and outward currents were blocked by a 5-HT(3) receptor antagonist, tropisetron, and a 5-HT(1A) receptor antagonist, NAN-190, respectively. The 5-HT-induced inward and outward currents were both associated with an increase in membrane conductance. The estimated reversal potential was more positive than -40 mV for the inward current and close to the calculated K(+) equilibrium potential for the outward current. The 5-HT application caused an increase, a decrease, or an increase followed by a decrease in the frequency of sIPSCs in pyramidal cells. The 5-HT(3) receptor agonist 1-(m chlorophenyl) biguanide increased the frequency of larger and fast-rising sIPSCs, whereas the 5-HT(1A) receptor agonist (+/-)8-hydroxydipropylaminotetralin hydrobromide elicited opposite effects and decreased the frequency of large events. These data indicate that serotonergic activation imposes complex actions on cortical inhibitory networks, which may lead to changes in cortical information processing. PMID- 12626612 TI - Vibration-induced changes in EMG during human locomotion. AB - The present study was set up to examine the contribution of Ia afferent input in the generation of electromyographic (EMG) activity. Subjects walked blindfolded along a walkway while tendon vibration was applied continuously to a leg muscle. The effects of vibration were measured on mean EMG activity in stance and swing phase. The results show that vibration of the quadriceps femoris (Q) at the knee and of biceps femoris (BF) at the knee enhanced the EMG activity of these muscles and this occurred mainly in the stance phase of walking. These results suggest involvement of Ia afferent input of Q and BF in EMG activation during stance. In contrast, vibration of muscles at the ankle and hip had no significant effect on burst amplitude. Additionally, the onset time of tibialis anterior was measured to look at timing of phase transitions. Only vibration of quadriceps femoris resulted in an earlier onset of tibialis anterior within the gait cycle, suggesting involvement of these Ia afferents in the triggering of phase transitions. In conclusion, the results of the present study suggest involvement of Ia afferent input in the control of muscle activity during locomotion in humans. A limited role in timing of phase transitions is proposed as well. PMID- 12626614 TI - Task-dependent modulation of multi-digit force coordination patterns. AB - When grasping and holding an object with five digits under a variety of task constraints, subjects use well-defined force coordination patterns, i.e., consistent force covariations and in-phase synchronization among all digit pairs. The question arises as to whether these force coordination patterns are default mechanisms for controlling multi-digit force production or whether they are specific to lifting and holding an object. To address this question, we asked subjects to grasp a manipulandum and exert forces with five digits simultaneously so as to match a force template measured from an actual object grasp, lift, and hold task (GLH). Unlike GLH, the force production task (FP) lacked the constraint of having to maintain object stability against gravity. The amplitude of individual finger forces and force covariations were similar for both tasks (with the exception of the little finger, which tended to produce less force in FP). Nonetheless, when multiple grip forces were not required to hold the manipulandum against gravity (FP), there was a significantly lower tendency for forces to be synchronized with higher intertrial variability of phase differences between forces exerted by all digit-pairs. Furthermore, the tendency for force phase differences to cluster at 0 degrees was lower for FP than GLH. These results suggest that some aspects of the control of multi-digit grasping, i.e., force synchronization, are specific to object lift and hold rather than to the production of multi-digit forces. Modeling work suggests that motor unit synchronization might play an important role in the modulation of force synchronization patterns. PMID- 12626615 TI - Corticosterone acts directly at the amygdala to alter spinal neuronal activity in response to colorectal distension. AB - Administration of glucocorticoids to the amygdaloid nucleus facilitates visceromotor responses to colorectal distension in rats. The aim of this study was to determine if colorectal hypersensitivity develops through central modulation of spinal neuronal activity. Stereotaxic delivery of corticosterone (n = 10) or cholesterol (control, n = 10) onto the dorsal margin of the amygdala was performed on male Fischer-344 rats. Seven days later, extracellular potentials of single L(6)-S(1) spinal neurons were examined for responses to colorectal distension (CRD, 20-80 mmHg, 20 s) in sodium pentobarbital anesthetized and paralyzed animals. The proportions of neurons that responded to noxious CRD in corticosterone-implanted (62/186, 33%) and cholesterol-implanted (55/163, 34%) animals were virtually identical. However, the mean excitatory response of spinal neurons to CRD in corticosterone-treated rats was significantly greater (26.7 +/- 2.2 vs. 16.4 +/- 1.8 imp/s, P < 0.01) and the duration was longer (37.0 +/- 3.9 vs. 25.8 +/- 1.5 s, P < 0.05) than in the control group. No significant differences were found in neural responses to nonnoxious and noxious mechanical stimulation of somatic fields between corticosterone-implanted and control groups. In conclusion, our data support the hypothesis that central stimulation of the amygdala by corticosterone sensitizes the lumbosacral spinal neurons that mediate visceromotor reflexes to CRD. PMID- 12626613 TI - Baseline glutamate levels affect group I and II mGluRs in layer V pyramidal neurons of rat sensorimotor cortex. AB - Possible functional roles for glutamate that is detectable at low concentrations in the extracellular space of intact brain and brain slices have not been explored. To determine whether this endogenous glutamate acts on metabotropic glutamate receptors (mGluRs), we obtained whole cell recordings from layer V pyramidal neurons of rat sensorimotor cortical slices. Blockade of mGluRs with (+)-alpha-amino-4-carboxy-alpha-methyl-benzeacetic acid (MCPG, a general mGluR antagonist) increased the mean amplitude of spontaneous excitatory postsynaptic currents (sEPSCs), an effect attributable to a selective increase in the occurrence of large amplitude sEPSCs. 2S-2-amino-2-(1S,2S-2-carboxycyclopropyl-1 yl)-3-(xanth-9-yl)propanoic acid (LY341495, a group II antagonist) increased, but R(-)-1-amino-2,3-dihydro-1H-indene-1,5-dicarboxylic acid (AIDA) and (RS)-hexyl HIBO (group I antagonists) decreased sEPSC amplitude, and (R,S)-alpha-cyclopropyl 4-phosphonophenylglycine (CPPG, a group III antagonist) did not change it. The change in sEPSCs elicited by MCPG, AIDA, and LY341495 was absent in tetrodotoxin, suggesting that it was action potential-dependent. The increase in sEPSCs persisted in GABA receptor antagonists, indicating that it was not due to effects on inhibitory interneurons. AIDA and (S)-3,5-dihydroxyphenylglycine (DHPG, a group I agonist) elicited positive and negative shifts in holding current, respectively. LY341495 and (2S,2'R,3'R)-2-(2',3'-dicarboxycyclopropyl)glycine (DCG-IV, a group II agonist) elicited negative and positive shifts in holding current, respectively. The AIDA and LY341495 elicited currents persisted in TTX. Finally, in current clamp, LY341495 depolarized cells by approximately 2 mV and increased the number of action potentials to a given depolarizing current pulse. Thus ambient levels of glutamate tonically activate mGluRs and regulate cortical excitability. PMID- 12626616 TI - Synaptic modulation of the interspike interval signatures of bursting pyloric neurons. AB - The pyloric network of the lobster stomatogastric nervous system is one of the best described assemblies of oscillatory neurons producing bursts of action potentials. While the temporal patterns of bursts have been investigated in detail, those of spikes have received less attention. Here we analyze the intraburst firing patterns of pyloric neurons and the synaptic interactions shaping their dynamics in millisecond time scales not performed before. We find that different pyloric neurons express characteristic, cell-specific firing patterns in their bursts. Nonlinear analysis of the interspike intervals (ISIs) reveals distinctive temporal structures ('interspike interval signatures'), which are found to depend on the synaptic connectivity of the network. We compare ISI patterns of the pyloric dilator (PD), lateral pyloric (LP), and ventricular dilator (VD) neurons in 1) normal conditions, 2) after blocking glutamatergic synaptic connections, and 3) in various functional configurations of the three neurons. Manipulation of the synaptic connectivity results in characteristic changes in the ISI signatures of the postsynaptic neurons. The intraburst firing pattern of the PD neuron is regularized by the inhibitory synaptic connection from the LP neuron as revealed in current-clamp experiments and also as reconstructed with a dynamic clamp. On the other hand, mutual inhibition between the LP and VD neurons tend to produce more irregular bursts with increased spike jitter. The results show that synaptic interactions fine-tune the output of pyloric neurons. The present data also suggest a way of processing of synaptic information: bursting neurons are capable of encoding incoming signals by altering the fine structure of their intraburst spike patterns. PMID- 12626617 TI - Intact synaptic GABAergic inhibition and altered neurosteroid modulation of thalamic relay neurons in mice lacking delta subunit. AB - Robust GABA-mediated inhibitory postsynaptic currents (IPSCs) in neurons of the thalamic relay (TC) nuclei are important in sustaining oscillatory activity within thalamic and thalamocortical circuits. The biophysical properties and pharmacological sensitivities of these IPSCs both depend on the subunit combination of postsynaptic gamma-aminobutyric acid-A (GABA(A)) receptors. Recombinant GABA(A) receptors containing the delta subunit (heavily expressed in TC nuclei) have been shown to exhibit slowed desensitization rates and high affinity for GABA in heterologous expression systems. We tested whether the GABA(A)-mediated synaptic inhibition in TC neurons would be affected by loss of the delta subunit. Spontaneous and evoked IPSCs were recorded from neurons in the ventral basal complex (VB) of the thalamus from brain slices of wild-type (delta(+/+)) and homozygous delta subunit deficient mice (delta(-/-)). Spontaneous IPSCs (sIPSCs) from delta(-/-) mice had no significant differences in amplitude, duration, or frequency compared with their delta(+/+) counterparts. However, baseline noise (63% of control) and the relative contribution of the slow component to overall decay (79% of control) were significantly lower in delta(-/-) VB recordings. Evoked IPSCs (eIPSCs) in delta(-/-) neurons showed no difference in peak amplitude, but had an accelerated slow decay component (40- vs. 55-ms time constant). We further tested whether neurosteroid modulation of GABA(A) receptors was dependent on the presence of the delta subunit, as previously reported in recombinant systems. Pregnenolone sulfate (PS) significantly reduced eIPSC peak amplitude (-30%) and increased duration in delta(-/-), but not in delta(+/+) mice. sIPSCs were not affected in any neurons, delta(-/-) or delta(+/+). In contrast, 3-alpha,5-alpha tetrahydrodeoxycorticosterone (THDOC) increased the durations of eIPSCs and sIPSCs in both delta(-/-) and delta(+/+) VB neurons. Our findings show that although the delta subunit confers a striking PS insensitivity to eIPSCs in VB neurons, it plays only a minor role in the synaptic inhibition of VB neurons. This suggests delta subunit containing GABA(A) receptors may be functionally limited to an extrasynaptic locus in VB neurons. PMID- 12626619 TI - Electrophysiological properties and input-output organization of callosal neurons in cat association cortex. AB - Intracellular recordings from association cortical areas 5 and 7 were performed in cats under barbiturate or ketamine-xylazine anesthesia to investigate the activities of different classes of neurons involved in callosal pathways, which were electrophysiologically characterized by depolarizing current steps. Excitatory postsynaptic potentials (EPSPs), inhibitory postsynaptic potentials (IPSPs), and/or antidromic responses were elicited by stimulating homotopic sites in the contralateral cortical areas. Differential features of EPSPs related to latencies, amplitudes, and slopes were detected in closely located (50 microm or less) neurons recorded in succession along the same electrode track. In contrast to synchronous thalamocortical volleys that excited most neurons within a cortical column, stimuli applied to homotopic sites in the contralateral cortex activated neurons at restricted cortical depths. Median latencies of callosally evoked EPSPs were 1.5 to 4 ms in various cortical cell-classes. Fast-rhythmic bursting neurons displayed EPSPs whose amplitudes were threefold larger, and latencies two- or threefold shorter, than those found in the three other cellular classes. Converging callosal and thalamic inputs were recorded in the same cortical neuron. EPSPs or IPSPs were elicited by stimulating foci spaced by <1 mm in the contralateral cortex. In the overwhelming majority of neurons, latencies of antidromic responses were between 1.2 and 3.1 ms; however, some callosal neurons had much longer latencies, 20 Hz was enhanced in cells from PV-/- mice, the largest difference between PV-/- and PV+/+ animals (220%) being observed at 33 Hz. The effect of IPSC facilitation at sustained gamma frequencies was assessed on kainate-induced rhythmic IPSC-paced neuronal oscillations at gamma frequencies, recorded with dual field potential recordings in area CA3. The maximum power of the oscillation was 138 microV(2) at 36 Hz in slices from PV+/+ mice and was trebled in slices from PV-/- mice. PV deficiency caused a similar increase in gamma power under conditions used to study IPSC facilitation and can be explained by an increased facilitation of GABA release at sustained high frequencies. The dominant frequency and coherence were not affected by PV deficiency. These observations suggest that PV deficiency, due to an increased short-term facilitation of GABA release, enhances inhibition by high-frequency burst-firing PV-expressing interneurons and may affect the higher cognitive functions associated with gamma oscillations. PMID- 12626621 TI - Responses to di-sodium guanosine 5'-monophosphate and monosodium L-glutamate in taste receptor cells of rat fungiform papillae. AB - The 5'-ribonucleotide guanosine 5'-monophosphate (GMP) is used widely as an umami taste stimulus and a potent flavor enhancer as it synergistically increases the umami taste elicited by monosodium glutamate. Transduction mechanisms for GMP and its synergy with glutamate are largely unknown. Using whole-cell patch-clamp and Ca(2+) imaging, we examined responses to GMP, glutamate, and a mixture of GMP and glutamate in taste-receptor cells of rat fungiform papillae. Our electrophysiological results showed that GMP induces responses that are similar to those of glutamate, e.g., an outward current, an inward current, or a biphasic response. Our Ca(2+) imaging results showed that applications of GMP, glutamate, and the mixture increased intracellular Ca(2+) levels. Interestingly, both patch clamp and Ca(2+) imaging showed that some taste cells can respond to GMP and glutamate independently, indicating that glutamate and GMP likely activate different receptors. Simultaneous application of GMP and glutamate resulted in synergistic responses in a subset of cells; both response intensity and number of responding cells were increased. Most responses to GMP, as well as the synergy between GMP and glutamate, were suppressed by 8-bromo-adenosine 3',5'-cyclic monophosphate (8-bromo-cAMP) in patch-clamp recordings. Together, our results suggest that intracellular cAMP- and Ca(2+)-mediated pathways are involved in umami taste transduction for GMP and its synergistic responses with glutamate. PMID- 12626622 TI - Response properties of TMJ units in superficial laminae at the spinomedullary junction of female rats vary over the estrous cycle. AB - Neurons responsive to stimulation of the temporomandibular joint (TMJ) region were recorded from superficial laminae at the trigeminal subnucleus caudalis/upper cervical cord (Vc/C(2)) junction region of cycling female rats under barbiturate anesthesia. To determine if receptive field (RF) properties or sensitivity to algesic chemicals of TMJ units vary over the estrous cycle, animals were selected from proestrous (high estrogen) or early diestrous (low estrogen) stages. More than 90% of TMJ units from each group received convergent nociceptive input [wide dynamic range (WDR) or nociceptive specific (NS)-like] from facial skin. The cutaneous high-threshold RF areas of WDR units from proestrous rats were 30% larger than diestrous units, while RF areas of NS units were similar. Bradykinin (BK, 0.1-10 microM) injection into the TMJ region excited a high percentage of units (>80% of total) from both groups in a dose related manner. However, BK-evoked response magnitude (R(mag), +140%) and duration (+64%) were greater for proestrous than diestrous units. Both WDR and NS like TMJ units of proestrous females displayed enhanced BK-evoked R(mag) values and response duration. Glutamate or mustard oil excitation of TMJ units was not affected by stage of the estrous cycle. Several TMJ units from proestrous and diestrous females were activated antidromically from the contralateral posterior thalamus, indicating that projection and nonprojection units were included in the sample population. These results were consistent with the hypothesis that factors related to stage of the estrous cycle modify the processing of deep craniofacial inputs by superficial dorsal horn neurons at the spinomedullary junction, a key region for the initial integration of sensory signals from the TMJ. PMID- 12626623 TI - Receptive field properties of human periodontal afferents responding to loading of premolar and molar teeth. AB - Impulses in 45 single mechanoreceptive afferents were recorded from the human inferior alveolar nerve with permucosally inserted tungsten microelectrodes. All afferents responded to mechanical stimulation of one or more premolar or molar teeth and most likely innervated their periodontal ligaments. For each afferent, isolated "ramp-and-hold" shaped force profiles of similar magnitudes (252 +/- 24 mN; mean +/- SD) were applied to the lower first premolar, the second premolar, and the first molar on the recording side. The tooth loads were applied in six directions: lingual, facial, mesial, and distal in the horizontal plane and up and down in the vertical direction of the tooth. The afferents response during the static phase of the stimulus was analyzed. All afferents were slowly adapting, discharging continuously in response to static forces in at least one stimulation direction. Twenty-nine afferents (64%) were spontaneously active, exhibiting an ongoing discharge in the absence of external stimulation. Stimulation of a single tooth was found to excite each afferent most strongly. The most sensitive tooth (MST) was the first premolar for 23, the second premolar for 13, and the first molar for 9 afferents. About half of the afferent population also responded to loading of one or two more teeth. The response profiles of these afferents indicated that the multiple-teeth receptive fields were due to mechanical coupling between the teeth rather than branching of single afferents to innervate several teeth. The afferent responses to loading the mesial and distal halves of the first molars were very similar. Thus both intensive and directional aspects of the afferent response when loading one side of the tooth was preserved to a great extent when loading the other side. When loading the MST, the afferents typically showed excitatory responses in two to four of the six stimulation directions, i.e., the afferents were broadly tuned to direction of tooth loading. In the horizontal plane, the afferent populations at the premolar teeth expressed no clear directional preferences. The afferents at the molar, however, showed a strong directional bias in the distal-lingual direction. In the vertical plane, there was a preference for downward-directed forces with a gradually decreasing sensitivity distally along the dental arch. The present results demonstrate that human periodontal afferents supplying anterior and posterior teeth differ in their capacity to signal horizontal and vertical forces, respectively. PMID- 12626624 TI - Multiple presynaptic and postsynaptic sites of inhibitory modulation by myomodulin at ARC neuromuscular junctions of Aplysia. AB - The functional activity of even simple cellular ensembles is often controlled by surprisingly complex networks of neuromodulators. One such network has been extensively studied in the accessory radula closer (ARC) neuromuscular system of Aplysia. The ARC muscle is innervated by two motor neurons, B15 and B16, which release modulatory peptide cotransmitters to shape ACh-mediated contractions of the muscle. Previous analysis has shown that key to the combinatorial ability of B15 and B16 to control multiple parameters of the contraction is an asymmetry in their peptide modulatory actions. B16, but not B15, releases myomodulin, which, among other actions, inhibits the contraction. Work in single ARC muscle fibers has identified a distinctive myomodulin-activated K current as a candidate postsynaptic mechanism of the inhibition. However, definitive evidence for this mechanism has been lacking. Here, working with the single fibers and then motor neuron-elicited excitatory junction potentials (EJPs) and contractions of the intact ARC muscle, we have confirmed two central predictions of the K-current hypothesis: the myomodulin inhibition of contraction is associated with a correspondingly large inhibition of the underlying depolarization, and the inhibition of both contraction and depolarization is blocked by 4-aminopyridine (4-AP), a potent and selective blocker of the myomodulin-activated K current. However, in the intact muscle, the experiments revealed a second, 4-AP-resistant component of myomodulin inhibition of both B15- and B16-elicited EJPs. This component resembles, and mutually occludes with, inhibition of the EJPs by another peptide modulator released from both B15 and B16, buccalin, which acts by a presynaptic mechanism, inhibition of ACh release from the motor neuron terminals. Direct measurements of peptide release showed that myomodulin also inhibits buccalin release from B15 terminals. At the level of contractions, nevertheless, the postsynaptic K-current mechanism is responsible for much of the myomodulin inhibition of peak contraction amplitude. The presynaptic mechanism, which is most evident during the initial build-up of the EJP waveform, underlies instead an increase of contraction latency. PMID- 12626625 TI - Somatotopic organization of the lateral part of area F2 (dorsal premotor cortex) of the macaque monkey. AB - The somatotopy of the lateral part of dorsal premotor area F2 has been studied by means of intracortical microstimulation and single neuron recording. The results show that most of this sector of F2 is excitable with low-intensity currents (3 40 microA) and that intracortical microstimulation evokes forelimb and trunk movements. Both proximal and distal forelimb movements are evoked in similar percentages. The proximal and distal forelimb representations partially overlap. However, proximal movements tend to be located more medially (laterally to the superior precentral dimple), whereas distal movements tend to be located more laterally (medially to the spur of the arcuate sulcus). The somatotopic organization demonstrated with microstimulation is confirmed by the similar somatotopic organization of active movements and of somatosensory properties revealed by single-neuron recording. The excitability and somatotopic organization of the lateral part of area F2 are discussed in relation to previous electrophysiological and anatomical findings. The involvement of the distal forelimb representation of area F2 in programming and controlling reaching to grasp movements is suggested. PMID- 12626626 TI - A corollary discharge mechanism modulates central auditory processing in singing crickets. AB - Crickets communicate using loud (100 dB SPL) sound signals that could adversely affect their own auditory system. To examine how they cope with this self generated acoustic stimulation, intracellular recordings were made from auditory afferent neurons and an identified auditory interneuron-the Omega 1 neuron (ON1) during pharmacologically elicited singing (stridulation). During sonorous stridulation, the auditory afferents and ON1 responded with bursts of spikes to the crickets' own song. When the crickets were stridulating silently, after one wing had been removed, only a few spikes were recorded in the afferents and ON1. Primary afferent depolarizations (PADs) occurred in the terminals of the auditory afferents, and inhibitory postsynaptic potentials (IPSPs) were apparent in ON1. The PADs and IPSPs were composed of many summed, small-amplitude potentials that occurred at a rate of about 230 Hz. The PADs and the IPSPs started during the closing wing movement and peaked in amplitude during the subsequent opening wing movement. As a consequence, during silent stridulation, ON1's response to acoustic stimuli was maximally inhibited during wing opening. Inhibition coincides with the time when ON1 would otherwise be most strongly excited by self generated sounds in a sonorously stridulating cricket. The PADs and the IPSPs persisted in fictively stridulating crickets whose ventral nerve cord had been isolated from muscles and sense organs. This strongly suggests that the inhibition of the auditory pathway is the result of a corollary discharge from the stridulation motor network. The central inhibition was mimicked by hyperpolarizing current injection into ON1 while it was responding to a 100 dB SPL sound pulse. This suppressed its spiking response to the acoustic stimulus and maintained its response to subsequent, quieter stimuli. The corollary discharge therefore prevents auditory desensitization in stridulating crickets and allows the animals to respond to external acoustic signals during the production of calling song. PMID- 12626627 TI - Electrophysiological classes of cat primary visual cortical neurons in vivo as revealed by quantitative analyses. AB - To facilitate the characterization of cortical neuronal function, the responses of cells in cat area 17 to intracellular injection of current pulses were quantitatively analyzed. A variety of response variables were used to separate the cells into subtypes using cluster analysis. Four main classes of neurons could be clearly distinguished: regular spiking (RS), fast spiking (FS), intrinsic bursting (IB), and chattering (CH). Each of these contained significant subclasses. RS neurons were characterized by trains of action potentials that exhibited spike frequency adaptation. Morphologically, these cells were spiny stellate cells in layer 4 and pyramidal cells in layers 2, 3, 5, and 6. FS neurons had short-duration action potentials (<0.5 ms at half height), little or no spike frequency adaptation, and a steep relationship between injected current intensity and spike discharge frequency. Morphologically, these cells were sparsely spiny or aspiny nonpyramidal cells. IB neurons typically generated a low frequency (<425 Hz) burst of spikes at the beginning of a depolarizing current pulse followed by a tonic train of action potentials for the remainder of the pulse. These cells were observed in all cortical layers, but were most abundant in layer 5. Finally, CH neurons generated repetitive, high-frequency (350-700 Hz) bursts of short-duration (<0.55 ms) action potentials. Morphologically, these cells were layer 2-4 (mainly layer 3) pyramidal or spiny stellate neurons. These results indicate that firing properties do not form a continuum and that cortical neurons are members of distinct electrophysiological classes and subclasses. PMID- 12626628 TI - Receptive field properties of unmyelinated tactile afferents in the human skin. AB - We recorded, with the microneurography technique, single-unit impulses from nine cutaneous mechanoreceptive afferents with conduction velocities in the C range and receptive fields in the hairy skin of the forearm. The units responded with high impulse rates to light touch and had low monofilament thresholds. The geography of receptive fields was explored with a scanning method: a lightweight probe with a small and rounded tip was made to scan the field area in a series of closely adjacent tracks while single-unit activity was recorded. The fields of the nine units varied considerably in size as well as complexity. The individual field consisted of one to nine small responsive spots distributed over an area of 1-35 mm(2) when explored with a moving indentation of 5 mN. The fields were roughly round or oval in shape with no preferred orientation. The size of the response differed between individual sensitive spots in a field, suggesting a highly nonuniform terminal organization. The properties of the fields seem consistent with a role of tactile C afferents to provide information about pleasant touch and skin-to-skin contacts to central structures controlling emotions and affiliative behavior. PMID- 12626629 TI - Spectral integration in A1 of awake primates: neurons with single- and multipeaked tuning characteristics. AB - We investigated modulations by stimulus components placed outside of the classical receptive field in the primary auditory cortex (A1) of awake marmosets. Two classes of neurons were identified using single tone stimuli: neurons with single-peaked frequency tuning characteristics (147/185, 80%) and neurons with multipeaked frequency tuning characteristics (38/185, 20%), referred to as single and multipeaked units, respectively. Each class of neurons was further studied using two-tone paradigms in which the frequency, intensity, and timing of the second tone were systematically varied while a unit was driven by the first tone placed at a unit's characteristic frequency (CF) if it was single-peaked or at one of multiple spectral peaks if it was multipeaked. The main findings were: 1) excitatory spectral peaks in the frequency tuning of the multipeaked units were often harmonically related. 2) Multipeaked units showed facilitation in their responses to combinations of two harmonically related tones placed at the spectral peaks of their frequency tuning. The two-tone facilitation was strongest for the simultaneously presented tones. 3) In 76 of 113 single-peaked units studied using the two-tone paradigm, facilitatory and/or inhibitory modulations by distant off-CF tones were observed. This distant inhibition differed from flanking (or side-band) inhibitions near CF. 4) In single-peaked units, the distant off-CF inhibitions were dominated by tones at frequencies that were harmonically related to the CF of a unit, whereas the facilitation by off-CF tones was observed for a wide range of frequencies. And 5) in both single- and multipeaked units, sound levels of two interacting tones determined whether the two tones produced excitation or inhibition. The largest facilitation was achieved by using two tones at their corresponding preferred sound levels. Together, these findings suggest that extracting or rejecting harmonically related components embedded in complex sounds may represent fundamental signal processing properties in different classes of A1 neurons. PMID- 12626630 TI - alpha 2-Adrenoceptor-mediated presynaptic modulation of GABAergic transmission in mechanically dissociated rat ventrolateral preoptic neurons. AB - The ventrolateral preoptic nucleus (VLPO) is a key nucleus involved in the homeostatic regulation of sleep-wakefulness. Little is known, however, about the cellular mechanisms underlying its role in sleep regulation and how the neurotransmitters, such as GABA and noradrenaline (NA), are involved. In the present study we investigated GABAergic transmission to acutely dissociated VLPO neurons using an enzyme-free, mechanical dissociation procedure in which functional terminals remained adherent and we investigated how this GABAergic transmission was modulated by NA. As previously reported in slices, NA hyperpolarized multipolar VLPO neurons and depolarized bipolar VLPO neurons. NA also inhibited the release of GABA onto multipolar VLPO neurons but had no effect on GABAergic transmission to bipolar neurons. The inhibition of release was mediated by presynaptic alpha(2) adrenoceptors coupled to N-ethylmaleimide (NEM) sensitive G-proteins which appeared to act via inhibition of adenylate cyclase and subsequent decreases in protein kinase A activity. The inhibition of GABA release did not, however, involve an inhibition of external Ca(2+) influx. The results indicate that all VLPO neurons contain GABAergic inputs and that the different morphological subgroups of VLPO neurons are correlated not only to different postsynaptic responses to NA but also to different presynaptic NA responses. Furthermore our results demonstrate an additional mechanism by which NA can modulate the excitability of multipolar VLPO neurons which may have important implications for its role in regulating sleep/wakefulness. PMID- 12626631 TI - Afferent innervation of the utricular macula in pigeons. AB - Biotinylated dextran amine (BDA) was used to retrogradely label afferents innervating the utricular macula in adult pigeons. The pigeon utriclar macula consists of a large rectangular-shaped neuroepithelium with a dorsally curved anterior edge and an extended medioposterior tail. The macula could be demarcated into several regions based on cytoarchitectural differences. The striola occupied 30% of the macula and contained a large density of type I hair cells with fewer type II hair cells. Medial and lateral extrastriola zones were located outside the striola and contained only type II hair cells. A six- to eight-cell-wide band of type II hair cells existed near the center of the striola. The reversal line marked by the morphological polarization of hair cells coursed throughout the epithelium, near the peripheral margin, and through the center of the type II band. Calyx afferents innervated type I hair cells with calyceal terminals that contained between 2 and 15 receptor cells. Calyx afferents were located only in the striola region, exclusive of the type II band, had small total fiber innervation areas and low innervation densities. Dimorph afferents innervated both type I and type II hair cells with calyceal and bouton terminals and were primarily located in the striola region. Dimorph afferents had smaller calyceal terminals with few type I hair cells, extended fiber branches with bouton terminals and larger innervation areas. Bouton afferents innervated only type II hair cells in the extrastriola and type II band regions. Bouton afferents innervating the type II band had smaller terminal fields with fewer bouton terminals and smaller innervation areas than fibers located in the extrastriolar zones. Bouton afferents had the most bouton terminals on the longest fibers, the largest innervation areas with the highest innervation densities of all afferents. Among all afferents, smaller terminal innervation fields were observed in the striola and large fields were located in the extrastriola. The cellular organization and innervation patterns of the utricular maculae in birds appear to represent an organ in adaptive evolution, different from that observed for amphibians or mammals. PMID- 12626632 TI - Abl family nonreceptor tyrosine kinases modulate short-term synaptic plasticity. AB - Abl family nonreceptor tyrosine kinases regulate cell morphogenesis through functional interactions with the actin cytoskeleton. The vertebrate Abl family kinases, Abl and Arg, are expressed in the adult mouse brain, where they may regulate actin cytoskeletal dynamics in mature neurons. Using immunoelectron microscopy, we have localized Abl and Arg to the pre- and postsynaptic compartments of synapses in the mouse hippocampal area CA1. Paired-pulse facilitation (PPF) was significantly reduced at the Schaffer collateral-CA1 (SC CA1) excitatory synapses in hippocampal slices from abl-/- and arg-/- mice as compared with wild-type mice. Furthermore, treatment of wild-type slices with the specific Abl family kinase inhibitor STI571 also reduced PPF. Basal synaptic transmission, posttetanic potentiation (PTP), long-term potentiation (LTP), and long-term depression (LTD) were similar to wild-type controls in abl-/- and arg-/ slices and in STI571-treated wild-type slices. These results indicate that an important function of Abl and Arg is to modulate synaptic efficacy via a presynaptic mechanism during repetitive activation. PMID- 12626634 TI - Independent neuronal oscillators of the rat globus pallidus. AB - In vivo, neurons of the globus pallidus (GP) and subthalamic nucleus (STN) resonate independently around 70 Hz. However, on the loss of dopamine as in Parkinson's disease, there is a switch to a lower frequency of firing with increased bursting and synchronization of activity. In vitro, type A neurons of the GP, identified by the presence of I(h) and rebound depolarizations, fire at frequencies ( (+)-(1R,2R)-U50,488 > lidocaine. The antinociception produced by lower doses of (-)-(1S,2S)-U50,488 was blocked by intrapaw nor-binaltorphimine as well as by antisense knockdown of the KOR. Such pretreatments, however, did not block the antinociception produced by (+)-(1R,2R) U50,488, lidocaine, or higher doses of (-)-(1S,2S)-U50,488. These data suggest that the sodium channel blocking effects of U50,488 and similar kappa-ORAs can contribute to their peripheral antinociceptive actions. PMID- 12626644 TI - Chronopharmacology of analgesic effect and its tolerance induced by morphine in mice. AB - The influence of morphine dosing time on analgesic effect after acute or chronic treatment, recovery of analgesic effect after once developed tolerance, and their pharmacological mechanisms were investigated in ICR male mice under a 12-h light/dark cycle (light on from 7:00 AM to 7:00 PM). There was a significant 24-h rhythm in the latency of thermal response at 30 min after morphine injection. The analgesic effect was significantly greater at the dark phase than at the light phase. The rhythmic pattern resembled overall the rhythm occurring in the latency of thermal response under non-drugged state. The absolute value of morphine analgesic effect (the real time spent on the hot-plate) on days 1 and 2 after morphine daily injection was significantly larger after morphine injection at 9:00 PM than after saline injection at 9:00 PM or after morphine injection at 9:00 AM. The recovery from tolerance of analgesic effect was significantly faster at the dark phase than at the light phase. The time-dependent difference in the analgesic effect after chronic treatment or recovery from tolerance is closely related to that in the expression of mu-opioid receptor. The present study suggests that 24-h rhythm of morphine analgesic effect is consistent with 24-h rhythm of mu-opioid receptor expression. PMID- 12626646 TI - Vasodilatory and electrophysiological actions of 8-iso-prostaglandin E2 in porcine coronary artery. AB - We examined the effects of several E-ring and F-ring isoprostanes on mechanical and electrophysiological activity in porcine coronary artery. Several isoprostanes evoked concentration-dependent contractions, with 8-iso-PGE2 being the most potent (-log EC50 of 6.9 +/- 0.1); this excitatory effect has been described in detail elsewhere and was not examined further here. 8-iso-PGE2 evoked dose-dependent relaxations in tissues preconstricted with the thromboxane A2-agonist U46619 (10(-6) M), with a negative log EC50 of 6.0 +/- 0.1 (n = 5). 8 iso-PGE1 and 8-iso-PGF2 beta also evoked relaxations (albeit with lower potency), whereas the other F-ring isoprostanes (8-iso-PGF1 alpha, 8-iso-PGF1 beta, and 8 iso-PGF2 alpha) were largely ineffective in this respect. The potency and efficacy of 8-iso-PGE2 in reversing tone were not dependent upon the concentration of U46619 used to preconstrict the tissues (10(-8) to 10(-6) M), indicating a lack of U46619-induced functional antagonism of these responses. 8 iso-PGE2 was able to completely relax tissues that had been denuded of endothelium (as indicated by loss of responsiveness to bradykinin). 8-iso-PGE2 evoked relaxations were markedly reduced by elevating the K+ equilibrium potential using 30 mM KCl and abolished by 60 mM KCl; they were also sensitive to charybdotoxin (10(-7) M) but not to 4-aminopyridine (1 mM). 8-iso-PGE2 also caused membrane hyperpolarization and augmentation of outward K+ current. We conclude that 8-iso-prostaglandin E2 acts directly on the smooth muscle to increase K+ conductance, leading to membrane hyperpolarization and vasodilation. PMID- 12626645 TI - A diarylheptanoid from lesser galangal (Alpinia officinarum) inhibits proinflammatory mediators via inhibition of mitogen-activated protein kinase, p44/42, and transcription factor nuclear factor-kappa B. AB - The diarylheptanoid 7-(4'-hydroxy-3'-methoxyphenyl)-1-phenylhept-4-en-3-one (HMP) is a naturally occurring phytochemical found in lesser galangal (Alpinia officinarum). In the present study, we have demonstrated the anti-inflammatory properties of this compound on mouse macrophage cell line (RAW 264.7) and human peripheral blood mononuclear cells (PBMCs) in vitro. Treatment of RAW 264.7 cells with HMP (6.25-25 microM) significantly inhibited lipopolysaccharide (LPS) stimulated nitric oxide (NO) production. This compound also inhibited the release of LPS-induced proinflammatory cytokines interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) from human PB-MCs in vitro. In addition, Western blotting and reverse transcription-polymerase chain reaction analysis demonstrated that HMP decreased LPS-induced inducible nitric-oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein and mRNA expression in RAW 264.7 cells. Furthermore, HMP treatment also reduced nuclear factor-kappa B (NF-kappa B) DNA binding induced by LPS in RAW 264.7 cells. To elucidate the molecular mechanism for inhibition of proinflammatory mediators by HMP (25 microM), we have studied the effect of HMP on LPS-induced p38 and p44/42 mitogen-activated protein kinase (MAPK). We observed that the phosphorylation of p44/42 MAPK in LPS stimulated RAW 264.7 cells was markedly inhibited by HMP, whereas activation of p38 MAPK was not affected. These results suggested that HMP from lesser galangal suppressed the LPS-induced production of NO, IL-1 beta, and TNF-alpha and expression of iNOS and COX-2 gene expression by inhibiting NF-kappa B activation and phosphorylation of p44/42 MAPK. PMID- 12626647 TI - GABAA receptor alpha 1 and beta 2 subunit null mutant mice: behavioral responses to ethanol. AB - Mice lacking either the alpha1 or beta 2 subunit of the GABAA receptor were tested for ethanol, saccharin, or quinine consumption, ethanol-conditioned place preference, ethanol-conditioned taste aversion, ethanol-simulated motor activity, and handling-induced seizures following chronic consumption of an ethanol liquid diet. The alpha1 null mutants showed decreased ethanol and saccharin consumption, increased aversion to ethanol, and a marked stimulation of motor activity after injection of ethanol. The beta 2 null mutants showed decreased consumption of saccharin and quinine, but not ethanol. Surprisingly, neither mutant showed marked changes in handling induced seizures before or after withdrawal of ethanol. The unique effects of deletion of these two GABAA receptor subunits on ethanol responses are discussed in terms of the distinct changes in different populations of GABAA receptors. PMID- 12626648 TI - Multiple differences in agonist and antagonist pharmacology between human and guinea pig histamine H1-receptor. AB - Species isoforms of histamine H2-, H3-, and H4-receptors differ in their pharmacological properties. The study aim was to dissect differences between the human H1R (hH1R) and guinea pig H1R (ghH1R). We coexpressed hH1R and gpH1R with regulators of G-protein signaling in Sf9 insect cells and analyzed the GTPase activity of Gq-proteins. Small H1R agonists showed similar effects at hH1R and gpH1R, whereas bulkier 2-phenylhistamines and histaprodifens were up to approximately 10-fold more potent at gpH1R than at hH1R. Most 2-phenylhistamines and histaprodifens were more efficacious at gpH1R than at hH1R. Several first generation H1R antagonists were approximately 2-fold, and arpromidine-type H1R antagonists up to approximately 10-fold more potent at gpH1R than at hH1R. [3H]Mepyramine competition binding studies confirmed the potency differences of the GTPase studies. Phe-153-->Leu-153 or Ile-433-->Val-433 exchange in hH1R (hH1R ->gpH1R) resulted in poor receptor expression, low [3H]mepyramine affinity, and functional inactivity. The Phe-153-->Leu-153/Ile-433-->Val-433 double mutant expressed excellently but only partially changed the pharmacological properties of hH1R. Small H1R agonists and 2-phenylhistamines interacted differentially with human and guinea pig H2R in terms of potency and efficacy, respectively. Our data show the following: 1) there are differences in agonist- and antagonist pharmacology of hH1R and gpH1R encompassing diverse classes of bulky ligands. These differences may be explained by higher conformational flexibility of gpH1R relative to hH1R; 2) Phe-153 and Ile-433 are critical for proper folding and expression of hH1R; and 3) H2R species isoforms distinguish between H1R agonists. PMID- 12626649 TI - Role of cyclooxygenase-1 and -2, phospholipase C, and protein kinase C in prostaglandin-mediated gastroprotection. AB - Oral administration of the nonselective cyclooxygenase (COX) inhibitor indomethacin (20 mg/kg), the COX-1 inhibitor 5-(4-chlorophenyl)-1-(4 methoxyphenyl)-3-trifluoromethylpyrazole (SC-560) (20 mg/kg), or the COX-2 inhibitor rofecoxib (1-20 mg/kg) antagonized the gastroprotective effects of 16,16-dimethyl-prostaglandin (PG) E2 (75 ng/kg p.o.) and 20% ethanol in rats. The effects of the COX inhibitors were reversed by the activator of ATP-sensitive potassium (KATP) channels cromakalim (0.3-0.5 mg/kg p.o.). The protective effects of 16,16-dimethyl-PGE2 and 20% ethanol were counteracted by the phospholipase C inhibitor 1-(6-((17 beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H pyrrole-2,5-dione (U-73122), but not its inactive analog 1-(6-((17 beta-3 methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-2,5-pyrrolidine-dione (U-73343) (1 mg/kg each i.v.). Likewise, the protein kinase C inhibitors chelerythrine (0.7 mg/kg i.v.) and staurosporine (3 microg/kg i.v.) inhibited gastroprotection. Effects of these enzyme inhibitors were not reversed by cromakalim. Submaximally effective doses of SC-560 (0.2 mg/kg p.o.) and rofecoxib (0.02 mg/kg p.o.) were additive and abolished the protection induced by 20% ethanol. The findings show that inhibition of COX-1 or COX-2 antagonizes not only adaptive gastroprotection by 20% ethanol but also the protective effect of exogenous PG in a cromakalimsensitive manner. Endogenous PG obviously add to the protective activity of exogenous PG. Gastroprotection by PG involves phospholipase C, protein kinase C, and KATP channels. Activation of KATP channels does not exert protection when the activity of phospholipase C or protein kinase C is suppressed. PMID- 12626650 TI - Nitric oxide production modulates cyclosporin A-induced distal renal tubular acidosis in the rat. AB - Cyclosporine A (CsA) causes distal renal tubular acidosis (dRTA) in humans and rodents. Because mice deficient in nitric-oxide (NO) synthase develop acidosis, we examined how NO production modulated H+ excretion during acid loading and CsA treatment in a rat model. Rats received CsA, L-arginine (L-Arg), or N omega-nitro L-arginine methyl ester (L-NAME), or combinations of CsA and L-NAME or L-Arg, followed by NH4Cl (acute acid load). In vehicle-treated rats, NH4Cl loading reduced serum and urine (HCO3-) and urine pH, which was associated with increases in serum [K+] and [Cl-] and urine NH3 excretion. Similar to CsA (7.5 mg/kg), L NAME impaired H+ excretion of NH4Cl-loaded animals. The combination CsA and L NAME reduced H+ excretion to a larger extent than either drug alone. In contrast, administration of L-Arg ameliorated the effect of CsA on H+ excretion. Urine pH after NH4Cl was 5.80 +/- 0.09, 6.11 +/- 0.13*, 6.37 +/- 0.16*, and 5.77 +/- 0.09 in the vehicle, CsA, CsA + L-NAME and CsA + L-Arg groups, respectively (*P < 0.05). The effect of CsA and alteration of NO synthesis were mediated at least in part by changes in bicarbonate absorption in perfused cortical collecting ducts. CsA or L-NAME reduced net HCO3- absorption, and, when combined, completely inhibited it. CsA + L-Arg restored HCO3- absorption to near control levels. Administration of CsA along with L-NAME reduced NO production to below levels observed with either drug alone. These results suggest that CsA causes dRTA by inhibiting H+ pumps in the distal nephron. Inhibition of NO synthesis may be one of the mechanisms underlying the CsA effect. PMID- 12626651 TI - Mechanism and implications of brown adipose tissue proliferation in rats and monkeys treated with the thiazolidinedione darglitazone, a potent peroxisome proliferator-activated receptor-gamma agonist. AB - Thiazolidinediones represent an established class of insulin sensitizing agents for treating noninsulin-dependent diabetes mellitus. Darglitazone, a thiazolidinedione approximately 200x more potent than ciglitazone, was evaluated in preclinical safety assessment studies using rats (1, 5, and 50 mg/kg/day) and cynomolgus monkeys (50, 75, and 100 mg/kg/day). Darglitazone was a potent adipogenic agent in rats, causing hyperplastic/hypertrophic changes and firmness of white and perirenal, dorsal thoracic (TBAT), and interscapular brown adipose tissue (BAT). Progressive changes in BAT size, morphology, firmness, and fatty acid composition preceded clinical signs of impaired respiration and the subsequent development of a dose-dependent, life-threatening hydrothorax. The characteristics of the pleural effusate were consistent with lymphatic fluid. These adverse effects were ameliorated/reversed upon drug withdrawal and were insulin-dependent since rats rendered totally insulinopenic by streptozotocin pretreatment did not develop TBAT changes or hydrothorax. Although the effects of darglitazone on BAT changes were consistent with enhanced sensitivity to endogenous glucocorticoids, adrenalectomy, and dietary dehydroepiandrosterone administration were without a protective effect. Treated monkeys also developed white and BAT hyperplasia/hypertrophy, peripheral edema, and hydrothorax-related morbidity/mortality. Both species developed reversible, dose-related reductions in red blood cell parameters and follicular atresia. Peripheral and pulmonary edema are purportedly a multifactorial process involving vasodilatation, increased endothelial permeability, and/or plasma volume expansion due to reduced renal sodium excretion. Moreover, profound alterations in TBAT hypertrophy/hyperplasia/firmness may lead to discrete hydrothorax by restricting normal thoracic lymphatic drainage. Similar effects on adipose tissue, hemodilution, and edema (peripheral and pulmonary) were observed clinically with darglitazone and/or several other structurally similar/dissimilar PPAR-gamma agonists. PMID- 12626652 TI - Suppression of cytochrome P450 3A protein levels by proteasome inhibitors. AB - We have previously reported that CYP3A cross-links with polyubiquitinated proteins in microsomes from nicardipine-treated rats in a process that is distinct from classical polyubiquitination. To further examine the role of the proteasome in CYP3A degradation, we investigated the effects of proteasome inhibitors lactacystin, MG132, proteasome inhibitor 1, and hemin in primary cultures of rat and human hepatocytes. With the exception of hemin, these agents increased the total pool of ubiquitinated proteins in microsomes isolated from rat hepatocytes, indicating that lactacystin, MG132, and proteasome inhibitor 1 effectively inhibited the proteasome in these cells. All four agents caused a reduction in the amount of the major approximately 55-kDa CYP3A band, opposite to what would be expected if the ubiquitin-proteasome pathway degraded CYP3A. Only hemin treatment caused an increase in high molecular mass (HMM) CYP3A bands. Because hemin treatment did not alter levels of ubiquitin in CYP3A immunoprecipitates, the HMM CYP3A bands formed in response to hemin treatment clearly were not due to proteasome inhibition. Rather, because hemin treatment also caused an increase in HMM CYP3A in the detergent-insoluble fraction of the 10,000g pellet, the HMM CYP3A seems to represent a large protein complex that is unlikely to primarily represent ubiquitination. PMID- 12626653 TI - Cytosolic Ca2+ and phosphoinositide hydrolysis linked to constitutively active alpha 1D-adrenoceptors in vascular smooth muscle. AB - In the present study, we analyzed changes in intracellular Ca2+ levels and inositol phosphate accumulation related to a population of alpha 1d-adrenoceptors in rat aorta resembling constitutively active receptors. Following intracellular Ca2+ store depletion by noradrenaline in Ca2+-free medium and removal of the agonist, restoration of extracellular Ca2+ induced four signals: a biphasic (transient and sustained) increase in [Ca2+]i, inositol phosphate accumulation, and a contractile response in the aorta. The transient increase in Ca2+, the inositol phosphate accumulation, and the contractile response were not observed in aortae incubated with prazosin or BMY 7378 [8-[2-[4-(2-methoxyphenyl)-1 piperazinyl]ethyl]-8-azaspiro[4.5]decane-7,9-dione] (a selective alpha 1d adrenoceptor ligand), relating the three signals to alpha 1d-adrenoceptor activity. In the presence of nimodipine, only the sustained increase in Ca2+ and the inositol phosphate accumulation were observed, relating both signals to calcium entry through L-channels. The four signals were abolished by Ni2+. In the rat tail artery, where alpha 1d-adrenoceptors are not functionally active, restoration of extracellular Ca2+ after store depletion induced only a sustained increase in [Ca2+]i without inositol phosphate accumulation nor contractile response. Taken together these results suggest that in the aorta, Ca2+ entry is required for the recovery of cytosolic calcium levels and the display of the membrane signals related to the constitutive activity of alpha 1d-adrenoceptors, i.e., inositol phosphate formation and Ca2+ entry through L-type channels, which maintains a contractile response once the agonist has been removed. PMID- 12626654 TI - Roles of necrosis, Apoptosis, and mitochondrial dysfunction in S-(1,2 dichlorovinyl)-L-cysteine sulfoxide-induced cytotoxicity in primary cultures of human renal proximal tubular cells. AB - S-(1,2-Dichlorovinyl)-L-cysteine (DCVC) is the penultimate nephrotoxic metabolite of the environmental contaminant trichloroethylene. Although metabolism of DCVC by the cysteine conjugate beta-lyase is the most studied bioactivation pathway, DCVC may also be metabolized by the flavin-containing monooxygenase (FMO) to yield DCVC sulfoxide (DCVCS). Renal cellular injury induced by DCVCS was investigated in primary cultures of human proximal tubular (hPT) cells by assessment of time- and concentration-dependent effects on cellular morphology, acute cellular necrosis, apoptosis, mitochondrial function, and cellular glutathione (GSH) status. Confluent hPT cells incubated with as little as 10 microM DCVCS for 24 h exhibited morphological changes, although at least 100 microM DCVCS was required to produce marked changes. Acute cellular necrosis did not occur until 48 h with at least 200 microM DCVCS, indicating that this is a high-dose, late response. The extent of necrosis was similar to that with DCVC. In contrast, apoptosis occurred as early as 1 h with as little as 10 microM DCVCS and the extent of apoptosis was much less than that with DCVC. Mitochondrial function was maintained with DCVCS concentrations up to 100 microM, consistent with hPT cells only being competent to undergo apoptosis at early time points and relatively low concentrations. Marked depletion (>50%) of cellular GSH content was only observed with 500 microM DCVCS. These results, combined with previous studies showing protection from DCVC-induced necrosis and apoptosis by the FMO inhibitor methimazole, suggest that formation of DCVCS plays a significant role in trichloroethylene-induced renal cellular injury in hPT cells. PMID- 12626655 TI - Ligand-selective activation of mu-oid receptor: demonstrated with deletion and single amino acid mutations of third intracellular loop domain. AB - The mechanism for the differential regulation of the mu-opioid receptor by agonists is investigated by identifying the receptor domains used to define the relative efficacies of three mu-opioid receptor-selective agonists: [D-Ala2,N-Me Phe4,Gly5-ol]-enkephalin (DAMGO), morphine, and [N-MePhe3,D-Pro4]-morphiceptin (PL017) to inhibit forskolin-stimulated intracellular cAMP production in human embryonic kidney 293 cells. This was accomplished by systematically deleting four to five amino acids clusters within the third intracellular loop of rat mu-opioid receptor, Arg258 to Arg280, followed by Ala substitution and scanning studies of the 276RRITR280 sequence, the putative G protein-coupling motif. Deletion of the four to five amino acid clusters resulted in differential effects on the affinities of the agonists and antagonists, and also on the potencies and coupling efficiencies of the three opioid agonists. Ala scanning studies of the 276RRITR280 sequence revealed also the differences between [D-Ala2,N-Me-Phe4,Gly5 ol]-enkephalin (DAMGO), morphine, and PL017. Substitution of Arg276 or Ile278 with Ala reduced the potency of DAMGO but not that of morphine PL017. Meanwhile, mutation of Thr279 to Ala increased the potencies of morphine and PL017 but not that of DAMGO. The I278A mutation decreased the DAMGO coupling efficiency but increased the PL017 coupling efficiency. The R280A mutation resulted in the increase in PL017 potency and coupling efficiency without altering those of DAMGO and morphine. Thus, these mutation studies suggested that the activation of mu opioid receptor and interaction between the critical domains such as RRITR within third intracellular loop and the G proteins are agonist-selective. PMID- 12626656 TI - Histamine H4 receptor mediates chemotaxis and calcium mobilization of mast cells. AB - The diverse physiological functions of histamine are mediated through distinct histamine receptors. Mast cells are major producers of histamine, yet effects of histamine on mast cells are currently unclear. The present study shows that histamine induces chemotaxis of mouse mast cells, without affecting mast cell degranulation. Mast cell chemotaxis toward histamine could be blocked by the dual H3/H4 receptor antagonist thioperamide, but not by H1 or H2 receptor antagonists. This chemotactic response is mediated by the H4 receptor, because chemotaxis toward histamine was absent in mast cells derived from H4 receptor-deficient mice but was detected in H3 receptor-deficient mast cells. In addition, Northern blot analysis showed the expression of H4 but not H3 receptors on mast cells. Activation of H4 receptors by histamine resulted in calcium mobilization from intracellular calcium stores. Both G alpha i/o proteins and phospholipase C (PLC) are involved in histamine-induced calcium mobilization and chemotaxis in mast cells, because these responses were completely inhibited by pertussis toxin and PLC inhibitor 1-[6-[[17 beta-3-methoxyestra-1,3,5 (10)-trien-17-yl]amino]hexyl] 1H-pyrrole-2,5-dione (U73122). In summary, histamine was shown to mediate signaling and chemotaxis of mast cells via the H4 receptor. This mechanism might be responsible for mast cell accumulation in allergic tissues. PMID- 12626657 TI - Pb2+ inhibition of sympathetic alpha 7-nicotinic acetylcholine receptor-mediated nitrergic neurogenic dilation in porcine basilar arteries. AB - Chronic exposure to inorganic lead (Pb2+) has been shown to facilitate peripheral vasoconstriction causing hypertension. Effect of lead on cerebral vascular function has not been reported. We have suggested in isolated porcine cerebral arteries that alpha 7-nicotinic acetylcholine receptors (alpha 7-nAChRs) on perivascular sympathetic nerves mediate calcium influx in these neurons, resulting in release of norepinephrine. The released norepinephrine then acts on presynaptic beta2-adrenoceptors located on the neighboring nitrergic nerve terminals, causing nitric oxide (NO) release and vasodilation. Because Pb2+ has been shown to inhibit alpha 7-nAChR-mediated responses in the central nervous system, effects of Pb2+ on alpha 7-nAChR-mediated nitrergic neurogenic dilation in isolated porcine basilar arteries and calcium influx in cultured superior cervical ganglion (SCG) cells of the pig were examined using in vitro tissue bath and confocal microscopic techniques. The results indicated that Pb2+ (but not Cd2+, Zn2+, or Al3+) in a concentration-dependent manner blocked relaxation of endothelium-denuded basilar arterial rings induced by nicotine (100 microM) and choline (1 mM) without affecting relaxation induced by sodium nitroprusside or isoproterenol. Furthermore, significant calcium influx in cultured SCG cells induced by choline and nicotine was attenuated specifically by Pb2+ with IC50 values comparable with those from tissue bath study. These results provide evidence supporting that lead is a likely antagonist for alpha 7-nAChRs that are found on postganglionic sympathetic adrenergic nerve terminals of SCG origin. Furthermore, these results indicate that lead can attenuate dilation of cerebral arteries by blocking sympathetic nerve-mediated release of NO from the perivascular nitrergic nerves. PMID- 12626658 TI - Calcium-dependent inhibition of synaptosomal serotonin transport by the alpha 2 adrenoceptor agonist 5-bromo-N-[4,5-dihydro-1H-imidazol-2-yl]-6-quinoxalinamine (UK14304). AB - Termination of serotonergic transmission is the function of the plasma membrane 5 hydroxytryptamine (serotonin, 5-HT) transporter (SERT), which is also a high affinity target in vivo for antidepressants, amphetamines, and cocaine. Studies show that SERT is regulated by protein kinase- and phosphataselinked pathways. In contrast, receptor-linked modulation of SERT is only minimally defined. Because noradrenergic stimulation is reported to influence 5-HT release, we explored possible presynaptic adrenoceptor-mediated regulation of SERT. In mouse forebrain synaptosomes, alpha2-adrenoceptor agonists, particularly 5-bromo-N-[4,5-dihydro 1H-imidazol-2-yl]-6-quinoxalinamine (UK14304), triggered a concentration- and time-dependent decrease in 5-HT transport. In contrast, 5-HT uptake was unaffected by pharmacological alpha1-adrenoceptor activation. Kinetically, UK14304 significantly decreased the apparent substrate affinity, Km without altering transport capacity, Vmax. At concentrations of UK14304 supporting maximal inhibition of SERT in synaptosomes, no effect on SERT in transfected cells was observed, suggesting that UK14304 acts indirectly to reduce SERT activity. The effect of UK14304 on 5-HT uptake was not shared by other Na+ and Cl -dependent transporters. UK14304-mediated inhibition of SERT function was yohimbine-sensitive, as was inhibition triggered by norepinephrine, and was abolished in the absence of added Ca2+. Moreover, UK14304 effects were attenuated by voltage-sensitive Ca2+ channel antagonists, consistent with a role for Ca2+ in UK14304 effects. In agreement with altered 5-HT transport activity in vitro, in vivo chronoamperometry studies revealed that UK14304 significantly prolonged 5-HT clearance. Our findings suggest that UK14304 modulates SERT function in vitro and in vivo via signaling pathways, possibly supported by an influx of Ca2+ through voltage-sensitive Ca2+ channels. PMID- 12626660 TI - Insulin mimetic action of synthetic phosphorylated peptide inhibitors of glycogen synthase kinase-3. AB - Glycogen synthase kinase-3 (GSK-3) was shown to be a key factor in attenuation of the cellular action of insulin. We speculated that inhibition of GSK-3 might have a potential therapeutic value in treatment of insulin resistance and type 2 diabetes. Here, we present a novel class of specific phosphorylated peptides inhibitors of GSK-3, which in sharp contrast to other protein kinase inhibitors that are ATP analogs, are substrate-competitive. We show that the GSK-3 peptide inhibitor activated glycogen synthase activity 2.5-fold in human embryonic kidney 293 cells, and increased glucose uptake in primary mouse adipocytes in the absence or presence of insulin compared with cells treated with two respective peptide controls. In addition, an i.p. administration of GSK-3 peptide inhibitor to normal or insulin-resistant obese C57BL/6J mice, improved their performance on glucose tolerance tests compared with control-treated animals. We present here a novel rational strategy for developing specific GSK-3 inhibitors and point toward GSK-3 as a promising therapeutic target in insulin resistance and type-2 diabetes. PMID- 12626659 TI - An assessment of the mechanistic differences between two integrin alpha 4 beta 1 inhibitors, the monoclonal antibody TA-2 and the small molecule BIO5192, in rat experimental autoimmune encephalomyelitis. AB - Integrin alpha 4 beta 1 plays an important role in inflammatory processes by regulating the migration of lymphocytes into inflamed tissues. Here we evaluated the biochemical, pharmacological, and pharmacodynamic properties and efficacy in experimental autoimmune encephalomyelitis (EAE), a model of multiple sclerosis, of two types of alpha 4 beta 1 inhibitors, the anti-rat alpha 4 monoclonal antibody TA-2 and the small molecule inhibitor BIO5192 [2(S)-[[1-(3,5-dichloro benzenesulfonyl)-pyrrolidine-2(S)-carbonyl]-amino]-4-[4-methyl-2(S)-(methyl-[2-[4 (3-o-tolyl-ureido)-phenyl]-acetyl]-amino)-pentanoylamino]-butyric acid]. TA-2 has been extensively studied in rats and provides a benchmark for assessing function. BIO5192 is a highly selective and potent (KD of <10 pM) inhibitor of alpha 4 beta 1. Dosing regimens were identified for both inhibitors, which provided full receptor occupancy during the duration of the study. Both inhibitors induced leukocytosis, an effect that was used as a pharmacodynamic marker of activity, and both were efficacious in the EAE model. Treatment with TA-2 caused a decrease in alpha 4 integrin expression on the cell surface, which resulted from internalization of alpha 4 integrin/TA-2 complexes. In contrast, BIO5192 did not modulate cell surface alpha 4 beta 1. Our results with BIO5192 indicate that alpha 4 beta 7 does not play a role in this model and that blockade of alpha 4 beta 1/ligand interactions without down-modulation is sufficient for efficacy in rat EAE. BIO5192 is highly selective and binds with high affinity to alpha 4 beta 1 from four of four species tested. These studies demonstrate that BIO5192, a novel, potent, and selective inhibitor of alpha 4 beta 1 integrin, will be a valuable reagent for assessing alpha 4 beta 1 biology and may provide a new therapeutic for treatment of human inflammatory diseases. PMID- 12626661 TI - Angiotensin II type 1 receptor antagonists inhibit basal as well as low-density lipoprotein and platelet-activating factor-stimulated human monocyte chemoattractant protein-1. AB - Monocyte chemoattractant protein-1 (MCP-1) is a potent chemotactic agent for monocytes and other cells and is thought to be involved in atherosclerosis, recruiting monocytes to the subendothelial space or to the site of inflammation. Angiotensin II has been demonstrated, at least in animal models, to stimulate MCP 1 expression. We investigated the effect of the angiotensin II type 1 (AT1) receptor antagonists irbesartan and losartan on MCP-1 production by freshly isolated human monocytes. Irbesartan and losartan inhibited basal MCP-1 production in a dose-dependent manner. Low-density lipoprotein (LDL) stimulated MCP-1 in a concentration-dependent manner, with 200 microg/ml LDL protein giving a 2-fold increase in MCP-1. Irbesartan and losartan dose dependently blocked LDL stimulated MCP-1. An angiotensin II type 2 receptor antagonist, S-(+)-1-([4 (dimethylamino)-3-methylphenyl]methyl)-5-(diphenylacetyl)-4,5,6,7-tetrahydro-1H imidazo(4,5-c)pyridine-6-carboxylic acid (PD123319), had no significant effect on basal MCP-1 levels or LDL-stimulated MCP-1. After noting homology between the AT1 receptor and the platelet-activating factor (PAF) receptor, we showed that irbesartan inhibited both [3H]PAF binding to human monocytes and carbamyl-PAF stimulation of MCP-1. However, irbesartan affinity for the PAF receptor was 700 times less than PAF, suggesting that there may be another mechanism for irbesartan inhibition of PAF-stimulated MCP-1. This is the first report showing that AT1 receptor antagonists inhibit basal as well as LDL- and PAF-stimulated MCP-1 production in freshly isolated human monocytes. PMID- 12626662 TI - Prevention of alterations in intestinal permeability is involved in zinc inhibition of acute ethanol-induced liver damage in mice. AB - Acute ethanol exposure causes liver injury in experimental animals, and accumulating evidence suggests that a major responsible factor for the pathogenesis is endotoxemia, which results from bacterial endotoxin leakage from the small intestine due to increased intestinal permeability under alcohol challenge. The purpose of this study was to examine whether zinc pretreatment would inhibit acute ethanol-induced liver injury through prevention of intestinal permeability changes. Male 129 SvPCJ mice were treated with three intragastric doses of ZnSO4 at 5 mg of zinc ion per kg each dosing prior to acute ethanol challenge with a single oral dose of 6 g/kg ethanol. The zinc treatment did not alter the elevation of serum concentrations of alcohol. The acute ethanol exposure caused an elevation in serum alanine aminotransferase levels as well as fatty liver and hepatic degenerative necrotic foci as determined by biochemical assay and histochemical analysis, respectively. A significant increase in liver tumor necrosis factor-alpha (TNF-alpha) levels was detected by enzyme-linked immunosorbent assay. These pathological effects correlated well with increases in serum endotoxin levels. Importantly, acute ethanol treatment caused significant damage to the small intestine as determined by morphological analysis of intestinal sections and permeability assay. These alcohol-induced hepatic pathological changes and TNF-alpha elevation were significantly inhibited in the zinc-pretreated animals. The inhibitory action of zinc on alcohol-induced liver damage and activation of inflammation was associated with zinc suppression of alcohol-induced intestinal permeability changes. These results thus demonstrate that zinc prevention of increased intestinal permeability is importantly involved in the inhibition of acute ethanol-induced liver damage in mice. PMID- 12626663 TI - AGI-1067: a multifunctional phenolic antioxidant, lipid modulator, anti inflammatory and antiatherosclerotic agent. AB - To explore the therapeutic efficacy and potential mechanisms of action of a new class of antiatherosclerotic drugs, AGI-1067 [mono[4-[[1-[[3,5-bis(1,1 dimethylethyl)-4-hydroxyphenyl]thio]-1-methylethyl]thio]-2,6-bis(1,1 dimethylethyl)phenyl] ester] (butanedioc acid) was tested in several animal models of atherosclerosis. AGI-1067, a novel phenolic antioxidant, was well tolerated in a 1-year study in hypercholesterolemic cynomolgus monkeys. It lowered low-density lipoprotein cholesterol (LDLc) by 41 and 90% at oral doses of 50 and 150 mg/kg, respectively and increased high-density lipoprotein cholesterol (HDLc) by 107% at the higher dose. In contrast, another phenolic antioxidant, probucol, had a modest LDLc-lowering effect (15% at 250 mg/kg) while decreasing HDLc (37% at 150 mg/kg). Histopathology of the aortas and coronary arteries revealed no atherosclerosis in the AGI-1067 (150 mg/kg) group and minimal-to moderate atherosclerosis in the vehicle and probucol (150 mg/kg) groups. AGI-1067 also inhibited atherosclerosis in LDL receptor-deficient (LDLr -/-) mice and apolipoprotein E-deficient (ApoE -/-) mice even in the absence of a lipid lowering effect. In LDLr -/- mice, AGI-1067 reduced aortic atherosclerosis by 49%. In ApoE -/- mice, AGI-1067 reduced atherosclerosis by 25, 41, and 49% in the arch, thoracic, and abdominal regions of the aorta. AGI-1067 also reduced vascular cell adhesion molecule-1 (VCAM-1) and monocyte chemoattractant protein-1 (MCP-1) mRNA levels in lungs of lipopolysaccharide-stimulated mice. At the cellular level, AGI-1067 inhibited tumor necrosis factor-alpha-inducible expression of VCAM-1, MCP-1, and E-selectin in human aortic endothelial cells (IC50 values = 6, 10, and 25 microM, respectively). These data show that AGI-1067 can inhibit atherosclerosis not only via its lipid-lowering effects but also by having direct anti-inflammatory effects on the vessel wall and suggest that it may be a novel therapeutic agent for coronary artery disease. PMID- 12626664 TI - Chronic administration of haloperidol and olanzapine attenuates ketamine-induced brain metabolic activation. AB - The fact that chronic administration of typical and atypical antipsychotic drugs is required for optimal therapeutic response suggests that drug-induced adaptive neurochemical changes contribute to their mechanism of action. In the present study, the effects of chronic and acute haloperidol and olanzapine were compared on ketamine-induced activation of select brain regions, as reflected by altered regional 14C-2-deoxyglucose (2-DG) uptake. Rats were injected once daily with haloperidol (1 mg/kg) or olanzapine (10 mg/kg) for 21 days, and 20 to 24 h after the final injection was challenged with saline or ketamine (25 mg/kg). The washout period was used to test the effects of chronic drug treatment without the influence of acute drug administration. In vehicle-treated rats, ketamine increased 2-DG uptake in select brain regions, including medial prefrontal cortex, nucleus accumbens, caudate putamen, stratum lacunosum-moleculare of hippocampus, and basolateral nucleus of the amygdala. This selective activation was attenuated by prior chronic treatment with both haloperidol and olanzapine. After acute treatment, olanzapine, but not haloperidol, blocked the ketamine induced activation of 2-DG uptake. These data suggest that both haloperidol and olanzapine can induce adaptive responses that counteract effects of ketamine. However, the differences observed in the acute effects of the two drugs in the ketamine challenge model suggest that different mechanisms could be responsible for their common chronic action of attenuating ketamine-induced brain metabolic activation. PMID- 12626665 TI - Mitochondria and oxygen sensing: fueling the controversy. PMID- 12626666 TI - Rotenone selectively occludes sensitivity to hypoxia in rat carotid body glomus cells. AB - Carotid body glomus cells release transmitters in response to hypoxia due to the increase of excitability resulting from inhibition of O2 -regulated K+ channels. However, the mechanisms involved in the detection of changes of O2 tension are unknown. We have studied the interaction between glomus cell O2 sensitivity and inhibition of the mitochondrial electron transport chain (ETC) in a carotid body thin slice preparation in which catecholamine release from intact single glomus cells can be monitored by amperometry. Inhibition of the mitochondrial ETC at proximal and distal complexes induces external Ca2+-dependent catecholamine secretion. At saturating concentration of the ETC inhibitors, the cellular response to hypoxia is maintained. However, rotenone, a complex I blocker, selectively occludes the responsiveness to hypoxia of glomus cells in a dose dependent manner. The effect of rotenone is mimicked by 1-methyl-4 phenylpyridinium ion (MPP+), an agent that binds to the same site as rotenone, but not by complex I inhibitors acting on different sites. In addition, the effect of rotenone is not prevented by incubation of the cells with succinate, a substrate of complex II. These data strongly suggest that sensitivity to hypoxia of carotid body glomus cells is not linked in a simple way to mitochondrial electron flow and that a rotenone (and MPP+)-sensitive molecule critically participates in acute oxygen sensing in the carotid body. PMID- 12626667 TI - Modulation of human ether-a-go-go-related K+ (HERG) channel inactivation by Cs+ and K+. AB - Unlike many other native and cloned K+ channels, human ether-a-go-go-related K+ (HERG) channels show significant Cs+ permeability with a PCs/PK (the permeability of Cs+ relative to that of K+) of 0.36 +/- 0.03 (n = 10). Here, we find that raising the concentration of external Cs+ (Cs+o) dramatically slows HERG channel inactivation without affecting activation. Replacement of 5 mM K+o by 135 mM Cs+o increased both inactivation and recovery time constants and shifted the mid-point of the steady-state inactivation curve by 25 mV in the depolarized direction (n = 6, P < 0.01). Raising [Cs+]o also modulated the voltage sensitivity of inactivation gating. With 130 mM Cs+i and 135 mM NMDG+o, the inactivation time constant decreased e-fold per 47.5 +/- 1.1 mV (n = 5), and when 20 mM Cs+ was added to the bath solution, the inactivation time constant decreased e-fold per 20.6 +/- 1.3 mV (n = 5, P < 0.01). A quantitative analysis suggests that Cs+o binds to a site in the pore that is influenced by the transmembrane electrical field, so that Cs+o-induced slowing of HERG inactivation is less prominent at strong depolarizations. K+o has effects that are similar to Cs+o and their effects were additive, suggesting Cs+o and K+o may share a common mechanism of action. The strong effects of Cs+ on inactivation but not on activation highlight the importance of ion and channel interactions during the onset of inactivation in the HERG channel. PMID- 12626668 TI - Angiotensin II stimulates hyperplasia but not hypertrophy in immature ovine cardiomyocytes. AB - Rat and sheep cardiac myocytes become binucleate as they complete the 'terminal differentiation' process soon after birth and are not able to divide thereafter. Angiotensin II (Ang II) is known to stimulate hypertrophic changes in rodent cardiomyocytes under both in vivo and in vitro conditions via the AT1 receptor and intracellular extracellular regulated kinase (ERK) signalling cascade. We sought to develop culture methods for immature sheep cardiomyocytes in order to test the hypothesis that Ang II is a hypertrophic agent in the immature myocardium of the sheep. We isolated fetal sheep cardiomyocytes and cultured them for 96 h, added Ang II and phenylephrine (PE) for 48 h, and measured footprint area and proliferation (5-bromo-2'-deoxyuridine (BrdU) uptake) separately in mono vs. binucleate myocytes. We found that neither Ang II nor PE changed the footprint area of mononucleated cells. PE stimulated an increase in footprint area of binucleate cells but Ang II did not. Ang II increased myocyte BrdU uptake compared to serum free conditions, but PE did not affect BrdU uptake. The MAP kinase kinase (MEK) inhibitor UO126 prevented BrdU uptake in Ang II-stimulated cells and prevented cell hypertrophy in PE-stimulated cells. This paper establishes culture methods for immature sheep cardiomyocytes and reports that: (1) Ang II is not a hypertrophic agent; (2) Ang II stimulates hyperplastic growth among mononucleate myocytes; (3) PE is a hypertrophic agent in binucleate myocytes; and (4) the ERK cascade is required for the proliferation effect of Ang II and the hypertrophic effect of PE. PMID- 12626669 TI - Intravenously administered hypocretin-1 alters brain amino acid release: an in vivo microdialysis study in rats. AB - We have reported that intravenous administration of hypocretin (Hcrt or orexin) reverses the symptoms of narcolepsy in genetically narcoleptic dogs. We have also reported that the onset of symptoms in canine genetic narcolepsy is accompanied by degenerative changes in forebrain regions, particularly the septal nucleus and amygdala. In the present in vivo microdialysis study we have investigated the effect of intravenous administration of Hcrt-1 (orexin-A) to anaesthetized rats on glutamate and GABA release in the amygdala, a region with moderate Hcrt innervation, and in the cerebellar cortex, a region with sparse or no Hcrt innervation. We found that intravenous Hcrt administration caused a marked (> 60 %) and sustained (> 50 min) increase in glutamate release within the amygdala, but no change in release in the cerebellar cortex. We did not detect a significant change in GABA release. When calcium-free artificial cerebrospinal fluid was used as the microdialysis perfusate, Hcrt-1 no longer produced an increase in glutamate release. Hcrt may act via the calcium-dependent regulation of glutamate release in certain nuclei of the central nervous system. PMID- 12626670 TI - 5-HT1A receptors increase excitability of spinal motoneurons by inhibiting a TASK 1-like K+ current in the adult turtle. AB - The modulatory effects of serotonin mediated by 5-HT1A receptors in adult spinal motoneurons were investigated by intracellular recordings in a slice preparation from the turtle. In current-clamp mode, activation of 5-HT1A receptors by 8-OH DPAT led to depolarization and an increase in input resistance in most motoneurons but caused hyperpolarization and a decrease in input resistance in the remaining smaller fraction of cells. When slices were preincubated in medium containing the 5-HT1A receptor antagonist WAY-100635, 8-OH-DPAT had no effect. In voltage-clamp mode, with 1 mM CsCl in the bathing medium, 8-OH-DPAT consistently inhibited a leak current that was sensitive to extracellular acidification and anandamide, a TASK-1 channel blocker. In medium with a low pH, as in the presence of anandamide, 8-OH-DPAT had no effect. Our results show that activation of 5 HT1A receptors contributes to the excitatory effect of serotonin on spinal motoneurons by inhibition of a TASK-1 potassium channel leading to depolarization and increased input resistance. PMID- 12626671 TI - The human respiratory gate. AB - Respiratory activity phasically alters membrane potentials of preganglionic vagal and sympathetic motoneurones and continuously modulates their responsiveness to stimulatory inputs. The most obvious manifestation of this 'respiratory gating' is respiratory sinus arrhythmia, the rhythmic fluctuations of electrocardiographic R-R intervals observed in healthy resting humans. Phasic autonomic motoneurone firing, reflecting the throughput of the system, depends importantly on the intensity of stimulatory inputs, such that when levels of stimulation are low (as with high arterial pressure and sympathetic activity, or low arterial pressure and vagal activity), respiratory fluctuations of sympathetic or vagal firing are also low. The respiratory gate has a finite capacity, and high levels of stimulation override the ability of respiration to gate autonomic responsiveness. Autonomic throughput also depends importantly on other factors, including especially, the frequency of breathing, the rate at which the gate opens and closes. Respiratory sinus arrhythmia is small at rapid, and large at slow breathing rates. The strong correlation between systolic pressure and R-R intervals at respiratory frequencies reflects the influence of respiration on these two measures, rather than arterial baroreflex physiology. A wide range of evidence suggests that respiratory activity gates the timing of autonomic motoneurone firing, but does not influence its tonic level. I propose that the most enduring significance of respiratory gating is its use as a precisely controlled experimental tool to tease out and better understand otherwise inaccessible human autonomic neurophysiological mechanisms. PMID- 12626672 TI - Molecular determinants of cAMP-mediated regulation of the Na+-Ca2+ exchanger expressed in human cell lines. AB - The cardiac Na+-Ca2+ exchanger (NCX1) is one of the major sarcolemmal Ca2+ transporters of cardiomyocytes. Structure-function studies suggest that beta adrenergic inhibition of NCX1, as reported for frog, but not mammalian hearts, may be associated with a unique splice variant of frog cardiac NCX1 where insertion of an extra exon completes the coding of a nucleotide binding P-loop. To test the involvement of the P-loop in cAMP-mediated regulation of NCX1 we used four stably transfected human cell lines (a previously established line of baby hamster kidney (BHK) cells and three new lines of human embryonic kidney (HEK) cells) expressing: (1) wild-type dog NCX1 (dog NCX1); (2) wild-type frog NCX1 (frog NCX1); (3) chimeric frog-dog NCX1 incorporating the completed P-loop from the frog NCX1 into the dog NCX1 sequence (frog/dog NCX1); and (4) a mutated frog NCX1 where a putative protein kinase A (PKA) site was disrupted by substitution of a single serine residue with glycine (S374G frog NCX1). Structural expression of these NCX1 constructs was confirmed using Western blot analysis of extracted proteins and immunofluorescence imaging. The NCX1-generated current (INa-Ca) was reliably measured in cells expressing dog (2.0 +/- 0.15 pA pF-1), frog (0.6 +/- 0.1 pA pF-1) and frog/dog (0.6 +/- 0.1 pA pF-1) NCX1, but less so in those expressing S374G frog NCX1 (0.3 +/- 0.1 pA pF-1). Addition of 100 microM 8 bromoadenosine 3',5' cyclic monophosphate (8-Br-cAMP) suppressed INa-Ca of frog and frog/dog NCX1 by 60-80 %. The suppression of INa-Ca was smaller and transient in cells expressing S374G frog NCX1, and absent in cells expressing dog NCX1. Intracellular Ca2+ (Ca2+i)-transients, activated by rapid withdrawal of Na+, were also downregulated in the frog and frog/dog NCX1 and to a smaller and transient extent in S374G frog NCX1. Our findings suggest that the suppressive effect of beta-adrenergic agonists requires the presence of the P-loop domain of the frog NCX1, and provide evidence that the putative PKA site, present in both dog and frog NCX1, might also be critical in the cAMP-mediated regulation of the exchanger. PMID- 12626673 TI - Effect of strength training on human patella tendon mechanical properties of older individuals. AB - This study investigated the effect of strength training on the mechanical properties of the human patella tendon of older individuals. Subjects were assigned to training (n = 9; age 74.3 +/- 3.5 years, body mass 69.7 +/- 14.8 kg and height 163.4 +/- 9.1 cm, mean +/- S.D.) and control (n = 9; age 67.1 +/- 2 years, body mass 73.5 +/- 14.9 kg and height 168.3 +/- 11.5 cm) groups. Strength training (two series of 10 repetitions at 80 % of five-repetition maximum) was performed three times per week for 14 weeks using leg extension and leg press exercises. Measurements of tendon elongation during a ramp isometric knee extension were performed before and after training and control periods in vivo using ultrasonography. Training caused a decreased tendon elongation and strain at all levels of force and stress (P < 0.01). Baseline tendon elongation and strain at maximal tendon load were 4.7 +/- 1.1 mm and 9.9 +/- 2.2 %, respectively (maximum force: 3346 +/- 1168 N; maximum stress: 40 +/- 11 MPa). After training, these values decreased to 2.9 +/- 1.2 mm and 5.9 +/- 2.4 % (P < 0.01), respectively (maximum force: 3555 +/- 1257 N; maximum stress: 42 +/- 11 MPa). Tendon stiffness increased by 65 % (2187 +/- 713 to 3609 +/- 1220 N mm-1; P < 0.05) and Young's modulus increased by 69 % (1.3 +/- 0.3 to 2.2 +/- 0.8 GPa; P < 0.01). As a result of these changes, the rate of torque development increased by 27 % (482.8 +/- 302.5 to 612.6 +/- 401 N m s-1; P < 0.01) following training. No significant changes occurred in any measured variables in the control group (P > 0.05). This study shows for the first time that strength training in old age increases the stiffness and Young's modulus of human tendons. This may reduce the risk of tendon injury in old age and has implications for contractile force production and the rapid execution of motor tasks. PMID- 12626674 TI - Veratridine block of rat skeletal muscle Nav1.4 sodium channels in the inner vestibule. AB - Veratridine (VTD) is an alkaloid toxin found in Liliaceae plants. VTD causes persistent opening of the voltage-gated Na+ channel and reduces its single channel conductance by 75 %. The mechanisms for these different VTD actions are unknown. Recent reports indicate that the VTD receptor aligns closely with the local anaesthetic (LA) receptor, which resides at D1S6, D3S6 and D4S6 of the Na+ channel alpha-subunit. To study this alignment, we created a mutant with cysteine substitutions at three S6 residues (rNav1.4-N434C/L1280C/F1579C). Under voltage clamp conditions, amitriptyline and bupivacaine remained as potent blockers of this mutant channel when expressed in human embryonic kidney cells, whereas VTD completely failed to cause persistent opening. Unexpectedly, VTD at 100 microM progressively blocked mutant currents by 90.4 +/- 1.6 % (n = 5), as assayed at 0.1 Hz for 15 min. This VTD block was reversed little during wash-off: approximately 70 % of mutant currents did not return in 30 min. An increase in channel opening either by repetitive pulses at 1 Hz or by the inhibition of the fast inactivation hastened the VTD block. Co-application of amitriptyline or bupivacaine, which targeted the LA receptor, prevented this VTD block. Our data suggest that (a) the VTD receptor and the LA receptor overlap extensively, (b) receptor-bound VTD lies in the inner vestibule, and (c) VTD blocks this mutant channel as a bona fide Na+ channel blocker. We propose that VTD likewise blocks the wild-type open Na+ channel, albeit partially, to decrease the unitary conductance and to stabilize the open conformation for persistent opening. PMID- 12626675 TI - Differential effects of catecholamines on vascular rings from ductus venosus and intrahepatic veins of fetal sheep. AB - Ductus venosus (DV) sparing means the maintenance of blood flow through the DV following reduction of liver venous blood supply during fetal hypoxia. The present study compared the reactions of the isthmic portion of the DV and intrahepatic veins (IHVs) to catecholamines in vitro. Vessel rings of 1 mm width and 3 mm diameter were obtained from 17 fetal sheep (88-136 days gestational age, median 120 days). The immunohistochemical examination of the DV and IHV was performed in eight cases using an antibody against alpha-smooth muscle actin and an antibody against alpha-adrenergic receptors. Five vessel rings of the DV in early gestation (median 95 days) did not respond to KCl-induced depolarisation. Force development in response to KCl of both vessel types increased with gestational age (P < 0.05). The IHV required 4.1 +/- 0.8 min (mean +/- S.E.M.) and the DV 14.5 +/- 4.0 min to reach the maximum tension in response to KCl, which was 5.0 +/- 4.0 mN in the IHV and 2.2 +/- 1.9 mN in the DV (n = 12, P < 0.05). The maximum forces developed in response to noradrenaline (norepinephrine; 42 microM, n = 9) and adrenaline (epinephrine; 100 microM, n = 12) were about sixfold higher in the IHV rings than in the DV rings (P < 0.05). The EC50 values of the DV and the IHV rings to noradrenaline were 5.9 +/- 1.3 microM and 5.0 +/- 1.3 microM, respectively (P = 0.03). The EC50 values of the adrenaline responses were 2.5 +/- 0.5 microM for the DV and 2.2 +/- 0.7 microM for the IHV (not significant). The alpha-adrenergic receptors were present in the well-structured media of IHVs, but were less distinctive in the wall of the DV. DV sparing can be attributed to an increased resistance of IHVs to catecholamines compared with the DV. The different responses can be explained by different anatomical and functional properties of the two vessel types. PMID- 12626676 TI - Aging attenuates the vestibulorespiratory reflex in humans. AB - Activation of the vestibular system changes ventilation in humans. The purpose of the present study was to investigate whether aging alters the vestibulorespiratory reflex in humans. Because aging attenuates the vestibulosympathetic reflex, it was hypothesized that aging would attenuate the vestibulorespiratory reflex. Changes in ventilation during engagement of the semicircular canals and/or the otolith organs were measured in fourteen young (26 +/- 1 years) and twelve older subjects (66 +/- 1 years). In young subjects, natural engagement of the semicircular canals and the otolith organs by head rotation increased breathing frequency during dynamic upright pitch at 0.25 Hz (15 cycles min-1) and 0.5 Hz (30 cycles min-1) (delta2 +/- 1 and delta4 +/- 1 breaths min-1, respectively; P < 0.05) and during dynamic upright roll (delta2 +/ 1 and delta4 +/- 1, respectively; P < 0.05). In older subjects, the only significant changes in breathing frequency occurred during dynamic pitch and roll at 0.5 Hz (delta2 +/- 1 and delta2 +/- 1 for pitch and roll, respectively). Stimulation of the horizontal semicircular canals by yaw rotation increased minute ventilation in young but not older subjects. Selective engagement of the otolith organs during static head-down rotation did not alter breathing frequency in either the young or older subjects. The results of this study indicate that the vestibulorespiratory reflex is attenuated in older humans, with greater vestibular stimulation needed to activate the reflex. PMID- 12626677 TI - Mechanical effect of muscle spindles in the canine external intercostal muscles. AB - High-frequency mechanical vibration of the ribcage increases afferent activity from external intercostal muscle spindles, but the effect of this procedure on the mechanical behaviour of the respiratory system is unknown. In the present study, we have measured the changes in external intercostal muscle length and the craniocaudal displacement of the ribs during ribcage vibration (40 Hz) in anaesthetized dogs. With vibration, external intercostal inspiratory activity increased by approximately 50 %, but the respiratory changes in muscle length and rib displacement were unaltered. A similar response was obtained after the muscles in the caudal segments of the ribcage were sectioned and the caudally oriented force exerted by these muscles on the rib was removed, thus suggesting that activation of external intercostal muscle spindles by vibration generates little tension. Prompted by this observation, we also examined the role played by the external intercostal muscle spindles in determining the respiratory displacement of the ribs during breathing against high inspiratory airflow resistances. Although resistances consistently elicited prominent reflex increases in external intercostal inspiratory activity, the normal inspiratory cranial displacement of the ribs was reversed into an inspiratory caudal displacement. Also, this caudal rib displacement was essentially unchanged after section of the external intercostal muscles, whereas it was clearly enhanced after denervation of the parasternal intercostals. These findings indicate that stretch reflexes in external intercostal muscles confer insufficient tension on the muscles to significantly modify the mechanical behaviour of the respiratory system. PMID- 12626678 TI - Mutations at the GABA receptor selectivity filter: a possible role for effective charges. AB - An important feature of ligand-gated ion channels is their exquisite ability to discriminate between ions. Still, little is known about the mechanisms underlying, or structural determinates of, this ability. We examined the structural elements underlying the ionic selectivity of rho1 GABA receptors expressed in Xenopus oocytes and human embryonic kidney cells using site-directed mutagenesis and two-electrode voltage-clamp or patch-clamp techniques. The wild type GABA receptor was chloride selective, with a small but significant permeability to potassium (PNa+ : PK+ : PCl- = 0 : 0.03 :1). Mutation of an alanine to glutamate at position 291 (thought to be located at the intracellular end of the second transmembrane domain), formed a channel that exhibited little discrimination among ions (0.70:0.87:1), while deletion of a neighbouring proline (290) was chloride selective, but had elevated cation permeabilities compared to the wild-type channel (0.12 : 0.14 : 1). Together, the two mutations (DeltaP290/A291E) caused a reversal of selectivity (2.72 : 3.59 : 1). We also examined the effects of neutralizing and reversing the charge of the adjacent, and highly conserved, arginine. Mutation of the neighbouring arginine to glutamate (R292E) increased the cation permeability similar to the DeltaP290/A291E double mutant (2.4 : 3.0 : 1), whereas neutral mutations at this position (R292M or R292C) retained chloride selectivity (0 : 0.11 : 1.0 and 0 : 0.14 : 1.0, respectively). Our experiments suggest that the effective charge near the presumed intracellular mouth of the pore is critical for ionic selectivity. PMID- 12626679 TI - Modulation of human vestibular-evoked postural responses by alterations in load. AB - The effects of body loading and unloading on human postural responses elicited by 1 mA bilateral, bipolar galvanic vestibular stimulation (GVS) were investigated. Subjects stood symmetrically, and in separate experiments were either loaded by 16, 33 and 50 % of their body weight with weights attached to the trunk, or unloaded by 10, 20 and 30 % using a whole-body harness that partially lifted the body but was free to translate horizontally. Randomised blocks of stimuli for each loading/unloading condition were compared to a non-loaded control condition. The rate of lateral reaction force development over the period 200-350 ms poststimulus increased in both legs with loading and decreased with unloading. The rate of force development was always larger from the leg on the side of cathodal stimulation. Vertical force responses were equal and opposite in the two legs, increasing on the side of the cathode and decreasing on the side of the anode. The rate of vertical force development over the period 200-350 ms after stimulus onset was increased with loading and decreased with unloading. In the frontal plane, the rate of head and trunk tilt in space was increased and decreased with loading and unloading, respectively. However, the relative rate of head tilt with respect to the trunk was not affected by loading conditions. These experiments provide further evidence that load-related afferent feedback influences the processing of vestibular information for the control of balance. PMID- 12626680 TI - Leucine biosynthesis in fungi: entering metabolism through the back door. AB - After exploring evolutionary aspects of branched-chain amino acid biosynthesis, the review focuses on the extended leucine biosynthetic pathway as it operates in Saccharomyces cerevisiae. First, the genes and enzymes specific for the leucine pathway are considered: LEU4 and LEU9 (encoding the alpha-isopropylmalate synthase isoenzymes), LEU1 (isopropylmalate isomerase), and LEU2 (beta isopropylmalate dehydrogenase). Emphasis is given to the unusual distribution of the branched-chain amino acid pathway enzymes between mitochondrial matrix and cytosol, on the newly defined role of Leu5p, and on regulatory mechanisms governing gene expression and enzyme activity, including new evidence for the metabolic importance of the regulation of alpha-isopropylmalate synthase by coenzyme A. Next, structure-function relationships of the transcriptional regulator Leu3p are addressed, defining its dual role as activator and repressor and discussing evidence in support of the self-masking model. Recent data pointing at a more extended Leu3p regulon are discussed. An overview of the layered controls of the extended leucine pathway is provided that includes a description of the newly recognized roles of Ilv5p and Bat1p in maintaining mitochondrial integrity. Finally, branched-chain amino acid biosynthesis and its regulation in other fungi are summarized, the question of leucine as metabolic signal is addressed, and possible directions of future research in this area are outlined. PMID- 12626682 TI - Separation, identification, and characterization of microorganisms by capillary electrophoresis. AB - The use of capillary electrophoresis (CE) for the analysis, identification, and characterization of microorganisms has been gaining in popularity. The advantages of CE, such as small sample requirements, minimal sample preparation, rapid and simultaneous analysis, ease of quantitation and identification, and viability assessment, make it an attractive technique for the analysis of microbial analytes. As this instrumental method has evolved, higher peak efficiencies have been achieved by optimizing CE conditions, such as pH, ionic strength, and polymer additive concentration. Experimental improvements have allowed better quantitation and more accurate results. Many practical applications of this technique have been investigated. Viability and identification of microbes can be accomplished in a single analysis. This is useful for evaluation of microbial analytes in consumer products. Diagnosis of microbe-based diseases is now possible, in some cases, without the need for culture methods. Microbe-molecule, virus-antibody, or bacteria-antibiotic interactions can be monitored using CE, allowing for the screening of possible drug candidates. Fermentation can be monitored using this system. This instrumental approach can be adapted to many different applications, including assessing the viability of sperm cells. Progress has been made in the development of microelectrophoresis instrumentation. These advances will eventually allow the development of small, dedicated devices for the rapid, repetitive analyses of specific microbial samples. Although these methods may never fully replace traditional approaches, they are proving to be a valuable addition to the collection of techniques used to analyze, quantitate, and characterize microbes. This review outlines the recent developments in this rapidly growing field. PMID- 12626684 TI - Two families of mechanosensitive channel proteins. AB - Mechanosensitive (MS) channels that provide protection against hypoosmotic shock are found in the membranes of organisms from the three domains of life: bacteria, archaea, and eucarya. Two families of ubiquitous MS channels are recognized, and these have been designated the MscL and MscS families. A high-resolution X-ray crystallographic structure is available for a member of the MscL family, and extensive molecular genetic, biophysical, and biochemical studies conducted in many laboratories have allowed postulation of a gating mechanism allowing the interconversion of a tightly closed state and an open state that controls transmembrane ion and metabolite fluxes. In contrast to the MscL channel proteins, which are of uniform topology, the much larger MscS family includes protein members with topologies that are predicted to vary from 3 to 11 alpha helical transmembrane segments (TMSs) per polypeptide chain. Sequence analyses reveal that the three C-terminal TMSs of MscS channel proteins are conserved among family members and that the third of these three TMSs exhibits a 20-residue motif that is shared by the channel-forming TMS (TMS 1) of the MscL proteins. We propose that this C-terminal TMS in MscS family homologues serves as the channel forming helix in a homooligomeric structure. The presence of a conserved residue pattern for the putative channel-forming TMSs in the MscL and MscS family proteins suggests a common structural organization, gating mechanism, and evolutionary origin. PMID- 12626683 TI - Cytokinesis in bacteria. AB - Work on two diverse rod-shaped bacteria, Escherichia coli and Bacillus subtilis, has defined a set of about 10 conserved proteins that are important for cell division in a wide range of eubacteria. These proteins are directed to the division site by the combination of two negative regulatory systems. Nucleoid occlusion is a poorly understood mechanism whereby the nucleoid prevents division in the cylindrical part of the cell, until chromosome segregation has occurred near midcell. The Min proteins prevent division in the nucleoid-free spaces near the cell poles in a manner that is beginning to be understood in cytological and biochemical terms. The hierarchy whereby the essential division proteins assemble at the midcell division site has been worked out for both E. coli and B. subtilis. They can be divided into essentially three classes depending on their position in the hierarchy and, to a certain extent, their subcellular localization. FtsZ is a cytosolic tubulin-like protein that polymerizes into an oligomeric structure that forms the initial ring at midcell. FtsA is another cytosolic protein that is related to actin, but its precise function is unclear. The cytoplasmic proteins are linked to the membrane by putative membrane anchor proteins, such as ZipA of E. coli and possibly EzrA of B. subtilis, which have a single membrane span but a cytoplasmic C-terminal domain. The remaining proteins are either integral membrane proteins or transmembrane proteins with their major domains outside the cell. The functions of most of these proteins are unclear with the exception of at least one penicillin-binding protein, which catalyzes a key step in cell wall synthesis in the division septum. PMID- 12626681 TI - Agrobacterium-mediated plant transformation: the biology behind the "gene jockeying" tool. AB - Agrobacterium tumefaciens and related Agrobacterium species have been known as plant pathogens since the beginning of the 20th century. However, only in the past two decades has the ability of Agrobacterium to transfer DNA to plant cells been harnessed for the purposes of plant genetic engineering. Since the initial reports in the early 1980s using Agrobacterium to generate transgenic plants, scientists have attempted to improve this "natural genetic engineer" for biotechnology purposes. Some of these modifications have resulted in extending the host range of the bacterium to economically important crop species. However, in most instances, major improvements involved alterations in plant tissue culture transformation and regeneration conditions rather than manipulation of bacterial or host genes. Agrobacterium-mediated plant transformation is a highly complex and evolved process involving genetic determinants of both the bacterium and the host plant cell. In this article, I review some of the basic biology concerned with Agrobacterium-mediated genetic transformation. Knowledge of fundamental biological principles embracing both the host and the pathogen have been and will continue to be key to extending the utility of Agrobacterium for genetic engineering purposes. PMID- 12626686 TI - Nutrition and prevention of type 2 diabetes. AB - In recent years, the prevalence of type 2 diabetes has increased alarmingly worldwide, giving diabetes the dimension of an epidemic. Striking parallel increases in the prevalence of obesity reflect the importance of body fatness as a contributing factor to diabetes incidence. Moreover, it has been estimated that up to 75% of the risk of type 2 diabetes is attributable to obesity. Recent clinical trials and observational epidemiologic studies demonstrate the efficacy of lifestyle changes, including decreased energy intake, decreased fat intake, and weight loss, as well as regular participation in physical activity, in improving insulin sensitivity (SI) and reducing the risk of diabetes. This review evaluates evidence of the effect of diet on insulin resistance, insulin secretion, and glucose tolerance, and reflects on directions for future work toward primary prevention of type 2 diabetes. PMID- 12626687 TI - Trophic and cytoprotective nutrition for intestinal adaptation, mucosal repair, and barrier function. AB - Intestinal epithelial cell turnover (proliferation, migration, differentiation, and apoptosis) and gut barrier functions are dynamic processes that are markedly affected by nutritional status, the route of feeding, and the adequacy of specific nutrients in the diet. Emerging studies are defining potential therapeutic roles for specific nutrients and diet-derived compounds (including arginine, glutamate, glutamine, glutathione, glycine, vitamin A, zinc, and specific lipids) in gut mucosal turnover, repair, adaptation after massive bowel resection, and barrier function. The role and regulation of endogenous bowel flora in generating short-chain fatty acids from diet-derived fiber and other diet-derived compounds and the effects of these agents on gut function are increasingly being elucidated. Results of these investigations should define new nutritional methods for trophic and cytoprotective effects on the intestine in conditions such as inflammatory bowel disease, malnutrition, and short bowel syndrome. PMID- 12626688 TI - GUGULIPID: a natural cholesterol-lowering agent. AB - The resin of the Commiphora mukul tree has been used in Ayurvedic medicine for more than 2000 years to treat a variety of ailments. Studies in both animal models and humans have shown that this resin, termed gum guggul, can decrease elevated lipid levels. The stereoisomers E- and Z-guggulsterone have been identified as the active agents in this resin. Recent studies have shown that these compounds are antagonist ligands for the bile acid receptor farnesoid X receptor (FXR), which is an important regulator of cholesterol homeostasis. It is likely that this effect accounts for the hypolipidemic activity of these phytosteroids. PMID- 12626689 TI - Physiology and molecular biology of dietary iron absorption. PMID- 12626690 TI - Different approaches to define individual amino acid requirements. AB - A full review of the strengths and limitations of the various methods used to define amino acid requirements is provided. The focus is on the recent development of carbon oxidation techniques such as indicator amino acid oxidation and 24-h amino acid balance to determine dietary indispensable (essential) amino acid needs in adults. All approaches depend on the change in a metabolic parameter in response to graded intake of the test amino acid. In humans, the within-subject variance is less than the between-subject variance, which has led to an appreciation of the need to study each subject across a range of intakes, above and below the mean requirement level. The data can then be analyzed using two-phase linear regression crossover and a precise population mean requirement can be determined. Several approaches have been used to define the variance of the mean requirement. Finally, a minimally invasive indicator amino acid oxidation model has been developed which allows the determination of dietary essential amino acid requirements in children and other vulnerable populations. PMID- 12626691 TI - Biologic mechanisms of the protective role of lutein and zeaxanthin in the eye. AB - The macular region of the primate retina is yellow in color due to the presence of the macular pigment, composed of two dietary xanthophylls, lutein and zeaxanthin, and another xanthophyll, meso-zeaxanthin. The latter is presumably formed from either lutein or zeaxanthin in the retina. By absorbing blue-light, the macular pigment protects the underlying photoreceptor cell layer from light damage, possibly initiated by the formation of reactive oxygen species during a photosensitized reaction. There is ample epidemiological evidence that the amount of macular pigment is inversely associated with the incidence of age-related macular degeneration, an irreversible process that is the major cause of blindness in the elderly. The macular pigment can be increased in primates by either increasing the intake of foods that are rich in lutein and zeaxanthin, such as dark-green leafy vegetables, or by supplementation with lutein or zeaxanthin. Although increasing the intake of lutein or zeaxanthin might prove to be protective against the development of age-related macular degeneration, a causative relationship has yet to be experimentally demonstrated. PMID- 12626692 TI - Challenges and approaches to reducing foodborne illness. AB - Microorganisms have successfully adapted to changes in food production, processing, and preservation techniques, resulting in a number of new and emerging foodborne pathogens and the re-emergence of organisms that have been problematic in the past. To protect public health, science must meet the challenges that result from the remarkable adaptability of foodborne pathogens. However, not all of the challenges of preventing foodborne illness reside in the realm of science. Food safety policy must evolve in response to new scientific understanding of hazards in the food supply and an ever-changing food processing industry. The laws, regulations, and organizations comprising the food safety system frequently lag behind current scientific knowledge of the risks posed by foodborne pathogens. Future systemic changes to enhance food safety will require better understanding of risks associated with specific pathogens occurring in the food supply and the costs and benefits of implementing mitigation strategies. PMID- 12626693 TI - Nutritional regulation of milk fat synthesis. AB - Certain diets cause a marked reduction in milk fat production in ruminants. Commonly referred to as milk fat depression (MFD), the mechanism involves an interrelationship between rumen microbial processes and tissue metabolism. Numerous theories to explain this interrelationship have been proposed and investigations offer little support for theories that are based on a limitation in the supply of lipogenic precursors. Rather, the basis involves alterations in rumen biohydrogenation of dietary polyunsaturated fatty acids and a specific inhibition of mammary synthesis of milk fat. The biohydrogenation theory proposes that under certain dietary conditions, typical pathways of rumen biohydrogenation are altered to produce unique fatty acid intermediates that inhibit milk fat synthesis. Trans-10, cis-12 conjugated linoleic acid (CLA) has been identified as one example that is correlated with the reduction in milk fat. Investigations with pure isomers have shown that trans-10, cis-12 CLA is a potent inhibitor of milk fat synthesis, and similar to diet-induced MFD, the mechanism involves a coordinated reduction in mRNA abundance for key enzymes involved in the biochemical pathways of fat synthesis. A more complete identification of these naturally produced inhibitors of fat synthesis and delineation of cellular mechanisms may offer broader opportunities for application and understanding of the regulation of lipid metabolism. PMID- 12626694 TI - Diet and nutrition in poor and minority communities in the United States 100 years ago. AB - Atwater and his colleagues began studying food consumption in the closing years of the nineteenth century and from the very start devoted much effort to collecting data from poor and minority households. This paper reviews some of the fruits of these labors, particularly from the standpoint of what they contribute toward a better historical understanding of American food habits and nutrition. It surveys dietary data from African American, Appalachian white, Mexican American, native-born poor, and immigrant households. These data shed light on several areas of historical concern, including rural versus urban nutrition, seasonal hunger, class disparities, and food-habit change. I suggest the economically and culturally diverse sample of dietary patterns that comes to us as a legacy of the Atwater era sets the stage for a history of American food habits considerably more sophisticated than has been realized to date. PMID- 12626695 TI - Cell migration in the forebrain. AB - The forebrain comprises an intricate set of structures that are required for some of the most complex and evolved functions of the mammalian brain. As a reflection of its complexity, cell migration in the forebrain is extremely elaborated, with widespread dispersion of cells across multiple functionally distinct areas. Two general modes of migration are distinguished in the forebrain: radial migration, which establishes the general cytoarchitectonical framework of the different forebrain subdivisions; and tangential migration, which increases the cellular complexity of forebrain circuits by allowing the dispersion of multiple neuronal types. Here, we review the cellular and molecular mechanisms underlying each of these types of migrations and discuss how emerging concepts in neuronal migration are reshaping our understanding of forebrain development in normal and pathological situations. PMID- 12626697 TI - Cell-cell signaling during synapse formation in the CNS. AB - Synapses join individual nerve cells into a functional network. Specific cell cell signaling events regulate synapse formation during development and thereby generate a highly reproducible connectivity pattern. The accuracy of this process is fundamental for normal brain function, and aberrant connectivity leads to nervous system disorders. However, despite the overall precision with which neuronal circuits are formed, individual synapses and synaptic networks are also plastic and can readily adapt to external stimuli or perturbations. In recent studies, several trans-synaptic signaling systems have been identified that can mediate various aspects of synaptic differentiation in the central nervous system. It appears that these individual pathways functionally cooperate, thereby generating robustness and flexibility, which ensure normal nervous system function. PMID- 12626696 TI - Brain representation of object-centered space in monkeys and humans. AB - Visuospatial cognition requires taking into account where things are relative to each other and not just relative to the viewer. Consequently it would make sense for the brain to form an explicit representation of object-centered and not just of ego-centered space. Evidence bearing on the presence and nature of neural maps of object-centered space has come from two sources: single-neuron recording in behaving monkeys and assessment of the visual abilities of human patients with hemispatial neglect. Studies of the supplementary eye field of the monkey have revealed that it contains neurons with object-centered spatial selectivity. These neurons fire when the monkey has selected, as target for an eye movement or attention, a particular location defined relative to a reference object. Studies of neglect have revealed that in some patients the condition is expressed with respect to an object-centered and object-aligned reference frame. These patients neglect one side of an object, as defined relative to its intrinsic midline, regardless of its location and orientation relative to the viewer. The two sets of observations are complementary in the sense that the loss of neurons, such as observed in the monkey, could explain the spatial distribution of neglect in these patients. PMID- 12626698 TI - Molecular approaches to spinal cord repair. AB - Axon growth inhibitors associated with myelin and the glial scar contribute to the failure of axon regeneration in the injured adult mammalian central nervous system (CNS). A number of these inhibitors, their receptors, and signaling pathways have been identified. These inhibitors can now be neutralized by a variety of approaches that point to the possibility of developing new therapeutic strategies to stimulate regeneration after spinal cord injury. PMID- 12626699 TI - The enzymatic basis of processivity in lambda exonuclease. AB - Lambda exonuclease is a highly processive 5'-->3' exonuclease that degrades double-stranded (ds)DNA. The single-stranded DNA produced by lambda exonuclease is utilized by homologous pairing proteins to carry out homologous recombination. The extensive studies of lambda biology, lambda exonuclease enzymology and the availability of the X-ray crystallographic structure of lambda exonuclease make it a suitable model to dissect the mechanisms of processivity. lambda Exonuclease is a toroidal homotrimeric molecule and this quaternary structure is a recurring theme in proteins engaged in processive reactions in nucleic acid metabolism. We have identified residues in lambda exonuclease involved in recognizing the 5' phosphate at the ends of broken dsDNA. The preference of lambda exonuclease for a phosphate moiety at 5' dsDNA ends has been established in previous studies; our results indicate that the low activity in the absence of the 5'-phosphate is due to the formation of inert enzyme-substrate complexes. By examining a lambda exonuclease mutant impaired in 5'-phosphate recognition, the significance of catalytic efficiency in modulating the processivity of lambda exonuclease has been elucidated. We propose a model in which processivity of lambda exonuclease is expressed as the net result of competition between pathways that either induce forward translocation or promote reverse translocation and dissociation. PMID- 12626700 TI - DmTTF, a novel mitochondrial transcription termination factor that recognises two sequences of Drosophila melanogaster mitochondrial DNA. AB - Using a combination of bioinformatic and molecular biology approaches a Drosophila melanogaster protein, DmTTF, has been identified, which exhibits sequence and structural similarity with two mitochondrial transcription termination factors, mTERF (human) and mtDBP (sea urchin). Import/processing assays indicate that DmTTF is synthesised as a precursor of 410 amino acids and is imported into mitochondria, giving rise to a mature product of 366 residues. Band-shift and DNase I protection experiments show that DmTTF binds two homologous, short, non-coding sequences of Drosophila mitochondrial DNA, located at the 3' end of blocks of genes transcribed on opposite strands. The location of the target sequences coincides with that of two of the putative transcription termination sites previously hypothesised. These results indicate that DmTTF is the termination factor of mitochondrial transcription in Drosophila. The existence of two DmTTF binding sites might serve not only to stop transcription but also to control the overlapping of a large number of transcripts generated by the peculiar transcription mechanism operating in this organism. PMID- 12626701 TI - Platinum cross-linking of adenines and guanines on the quadruplex structures of the AG3(T2AG3)3 and (T2AG3)4 human telomere sequences in Na+ and K+ solutions. AB - The quadruplex structures of the human telomere sequences AG3(T2AG3)3 I and (T2AG3)4 II were investigated in the presence of Na+ and K+ ions, through the cross-linking of adenines and guanines by the cis- and trans [Pt(NH3)2(H2O)2](NO3)2 complexes 1 and 2. The bases involved in chelation of the cis- and trans-Pt(NH3)2 moieties were identified by chemical and 3'-exonuclease digestions of the products isolated after denaturing gel electrophoresis. These are the four adenines of each sequence and four out of the 12 guanines. Two largely different structures have been reported for I: A from NMR data in Na+ solution and B from X-ray data of a K+-containing crystal. Structure A alone agrees with our conclusions about the formation of the A1-G10, A13-G22, A1-A13 platinum chelates at the top of the quadruplex and A7-A19, G4-A19 and A7-G20 at the bottom, whether the Na+ or K+ ion is present. At variance with a recent proposal that structures A and B could be the major species in Na+ and K+ solutions, respectively, our results suggest that structure A exists predominantly in the presence of both ions. They also suggest that covalent platinum cross-linking of a human telomere sequence could be used to inhibit telomerase. PMID- 12626702 TI - Evolution of monoblepharidalean fungi based on complete mitochondrial genome sequences. AB - We have determined the complete mitochondrial DNA (mtDNA) sequences of three chytridiomycete fungi, Monoblepharella15, Harpochytrium94 and Harpochytrium105. Our phylogenetic analysis based on concatenated mitochondrial protein sequences confirms the placement of Mono blepharella15 together with Harpochytrium spp. and Hyaloraphidium curvatum within the taxonomic order Monoblepharidales, with overwhelming support. These four mtDNA sequences encode the standard fungal mitochondrial gene complement and, like certain other chytridiomycete fungi, encode a reduced complement of 7-9 tRNAs, some of which require 5'-tRNA editing to be functional. Highly conserved sequence elements were identified upstream of almost all protein-coding genes in the mtDNAs of Monoblepharella15 and both Harpochytrium species. Finally, a guanosine residue is conserved upstream of the predicted ATG or GTG start codons of almost every protein-coding gene in these genomes. The appearance of this G residue correlates with the presence of a non canonical cytosine residue at position 37 in the anticodon loop of the mitochondrial initiator tRNAs. Based on the unorthodox features in these four genomes, we propose that a 4 bp interaction between the CAUC anticodon of these tRNAs and GAUG/GGUG codons is involved in translation initiation in monoblepharidalean mitochondria. Intriguingly, a similar interaction may also be involved in mitochondrial translation initiation in the sea anemone Metridium senile. PMID- 12626703 TI - Activation of influenza virus RNA polymerase by the 5' and 3' terminal duplex of genomic RNA. AB - The current model for influenza virus mRNA transcription involves the sequential interaction of the viral polymerase with the 5'- and 3'-ends of vRNA, with each RNA-protein interaction triggering a polymerase function necessary for cap-primed transcription. Here we show that the order in which this ternary complex is assembled is in fact important. Polymerase bound simultaneously to a pre-annealed duplex of the 5'- and 3'-ends of vRNA had greatly increased levels of primer binding and endonuclease activities compared to a sequentially assembled complex. Increased primer binding was due to the activation of a high affinity binding site with a preference for primer length RNAs. This correlated with enhanced levels of cap-primed transcription. Polymerase that was bound initially to just 5' vRNA had low primer binding activity, but was endonucleolytically active. Neither activity was significantly increased by the subsequent addition of 3' vRNA, and this sequentially assembled complex had correspondingly low mRNA transcription activity. Nevertheless, both routes of assembly led to complexes that were highly competent for dinucleotide ApG-primed transcription. Therefore, polymerase complexes assembled on pre-annealed 5' and 3' terminal viral RNA sequences have distinct properties from those assembled by sequential loading of polymerase onto the 5'-end followed by the 3'-end. This suggests a mechanism by which the virus couples transcription initiation and termination during mRNA transcription. PMID- 12626704 TI - Crystal structure of the Escherichia coli dcm very-short-patch DNA repair endonuclease bound to its reaction product-site in a DNA superhelix. AB - Very-short-patch repair (Vsr) enzymes occur in a variety of bacteria, where they initiate nucleotide excision repair of G:T mismatches arising by deamination of 5 methyl-cytosines in specific regulatory sequences. We have now determined the structure of the archetypal dcm-Vsr endonuclease from Escherichia coli bound to the cleaved authentic hemi-deaminated/hemi-methylated dcm sequence 5'-C-OH-3' 5' p-T-p-A-p-G-p-G-3'/3'-G-p-G-p-T-p(Me5)C-p-C formed by self-assembly of a 12mer oligonucleotide into a continuous nicked DNA superhelix. The structure reveals the presence of a Hoogsteen base pair within the deaminated recognition sequence and the substantial distortions of the DNA that accompany Vsr binding to product sites. PMID- 12626705 TI - Human mitochondrial DNA is packaged with TFAM. AB - Mitochondrial transcription factor A (TFAM), a member of the high mobility group proteins, is essential for maintenance of mitochondrial DNA (mtDNA). Most TFAM and mtDNA (both of which are normally soluble) was recovered from the particulate fraction of human placental mitochondria when extracted with the non-ionic detergent Nonidet P-40. mtDNA and TFAM were co-immunoprecipitated by anti-TFAM antibodies. TFAM was released into the supernatant by DNase I digestion of mtDNA in the particulate fraction. Thus, TFAM and mtDNA are tightly associated with each other, and it is likely that few TFAM or mtDNA molecules exist in an unbound form in mitochondria. Based on the fact that TFAM is abundant enough to wrap mtDNA entirely, these results suggest that human mtDNA is packaged with TFAM. PMID- 12626706 TI - Yeast telomerase is specialized for C/A-rich RNA templates. AB - Telomeres, the protective caps of eukaryotic chromosomes, are maintained by the enzyme telomerase. This telomere-specific reverse transcriptase (RT) uses a small region of its RNA subunit as template to synthesize telomeric DNA, which is generally G/T rich in the strand that contains the 3' end. To further our understanding of why telomeres are usually G/T rich, we screened Saccharomyces cerevisiae telomerase RNA (TLC1) libraries with randomized template sequences for complementation of a tlc1 deletion and decapping of existing telomeres. Surprisingly, the vast majority of the 60 000 different mutant telomerase templates tested showed no activity in vivo. This deficiency was not due to impaired assembly with the catalytic subunit (Est2p) nor could it be alleviated by enforced telomerase recruitment to the telomeres. Rather, the mutant templates reduced the nucleotide addition processivity of telomerase. The functional RNA template sequences recovered in our screens preferentially contained two or more consecutive rC nucleotides, reminiscent of the wild-type template. Thus, in contrast to retroviral RTs that can reverse transcribe any RNA sequence into DNA, the budding yeast telomerase RT is specialized for its C-rich RNA template. PMID- 12626707 TI - Targeted and random bacterial gene disruption using a group II intron (targetron) vector containing a retrotransposition-activated selectable marker. AB - Mobile group II introns have been used to develop a novel class of gene targeting vectors, targetrons, which employ base pairing for DNA target recognition and can thus be programmed to insert into any desired target DNA. Here, we have developed a targetron containing a retrotransposition-activated selectable marker (RAM), which enables one-step bacterial gene disruption at near 100% efficiency after selection. The targetron can be generated via PCR without cloning, and after intron integration, the marker gene can be excised by recombination between flanking Flp recombinase sites, enabling multiple sequential disruptions. We also show that a RAM-targetron with randomized target site recognition sequences yields single insertions throughout the Escherichia coli genome, creating a gene knockout library. Analysis of the randomly selected insertion sites provides further insight into group II intron target site recognition rules. It also suggests that a subset of retrohoming events may occur by using a primer generated during DNA replication, and reveals a previously unsuspected bias for group II intron insertion near the chromosome replication origin. This insertional bias likely reflects at least in part the higher copy number of origin proximal genes, but interaction with the replication machinery or other features of DNA structure or packaging may also contribute. PMID- 12626708 TI - Mosquito has a single multisubstrate deoxyribonucleoside kinase characterized by unique substrate specificity. AB - In mammals four deoxyribonucleoside kinases, with a relatively restricted specificity, catalyze the phosphorylation of the four natural deoxyribonucleosides. When cultured mosquito cells, originating from the malaria vector Anopheles gambiae, were examined for deoxyribonucleoside kinase activities, only a single enzyme was isolated. Subsequently, the corresponding gene was cloned and over-expressed. While the mosquito kinase (Ag-dNK) phosphorylated all four natural deoxyribonucleosides, it displayed an unexpectedly higher relative efficiency for the phosphorylation of purine versus pyrimidine deoxyribonucleosides than the fruit fly multisubstrate deoxyribonucleoside kinase (EC 2.7.1.145). In addition, Ag-dNK could also phosphorylate some medically interesting nucleoside analogs, like stavudine (D4T), 2-chloro-deoxyadenosine (CdA) and 5-bromo-vinyl-deoxyuridine (BVDU). Although the biological significance of multisubstrate deoxyribonucleoside kinases and their diversity among insects remains unclear, the observed variation provides a whole range of applications, as species specific and highly selective targets for insecticides, they have a potential to be used in the enzymatic production of various (di-)(deoxy-)ribonucleoside monophosphates, and as suicide genes in gene therapy. PMID- 12626709 TI - Complete sequence of the mitochondrial genome of Tetrahymena thermophila and comparative methods for identifying highly divergent genes. AB - The complete sequence of the mitochondrial genome of Tetrahymena thermophila has been determined and compared with the mitochondrial genome of Tetrahymena pyriformis. The sequence similarity clearly indicates homology of the entire T.thermophila and T.pyriformis mitochondrial genomes. The T.thermophila genome is very compact, most of the intergenic regions are short (only three are longer than 63 bp) and comprise only 3.8% of the genome. The nad9 gene is tandemly duplicated in T.thermophila. Long terminal inverted repeats and the nad9 genes are undergoing concerted evolution. There are 55 putative genes: three ribosomal RNA genes, eight transfer RNA genes, 22 proteins with putatively assigned functions and 22 additional open reading frames of unknown function. In order to extend indications of homology beyond amino acid sequence similarity we have examined a number of physico-chemical properties of the mitochondrial proteins, including theoretical pI, molecular weight and particularly the predicted transmembrane spanning regions. This approach has allowed us to identify homologs to ymf58 (nad4L), ymf62 (nad6) and ymf60 (rpl6). PMID- 12626710 TI - Dihydropyrimidine amidohydrolases and dihydroorotases share the same origin and several enzymatic properties. AB - Slime mold, plant and insect dihydropyrimidine amidohydrolases (DHPases, EC 3.5.2.2), which catalyze the second step of pyrimidine and several anti-cancer drug degradations, were cloned and shown to functionally replace a defective DHPase enzyme in the yeast Saccharomyces kluyveri. The yeast and slime mold DHPases were over-expressed, shown to contain two zinc ions, characterized for their properties and compared to those of the calf liver enzyme. In general, the kinetic parameters varied widely among the enzymes, the mammalian DHPase having the highest catalytic efficiency. The ring opening was catalyzed most efficiently at pH 8.0 and competitively inhibited by the reaction product, N-carbamyl-beta alanine. At lower pH values DHPases catalyzed the reverse reaction, the closing of the ring. Apparently, eukaryote DHPases are enzymatically as well as phylogenetically related to the de novo biosynthetic dihydroorotase (DHOase) enzymes. Modeling studies showed that the position of the catalytically critical amino acid residues of bacterial DHOases and eukaryote DHPases overlap. Therefore, only a few modifications might have been necessary during evolution to convert the unspecialized enzyme into anabolic and catabolic ones. PMID- 12626712 TI - A revised annotation and comparative analysis of Helicobacter pylori genomes. AB - Huge amounts of genomic information are currently being generated. Therefore, biologists require structured, exhaustive and comparative databases. The PyloriGene database (http://genolist.pasteur.fr/PyloriGene) was developed to respond to these needs, by integrating and connecting the information generated during the sequencing of two distinct strains of Helicobacter pylori. This led to the need for a general annotation consensus, as the physical and functional annotations of the two strains differed significantly in some cases. A revised functional classification system was created to accommodate the existing data and to make it possible to classify coding sequences (CDS) into several functional categories to harmonize CDS classification. The annotation of the two complete genomes was revised in the light of new data, allowing us to reduce the percentage of hypothetical proteins from approximately 40 to 33%. This resulted in the reassignment of functions for 108 CDS (approximately 7% of all CDS). Interestingly, the functions of only approximately 13% of CDS (222 out of 1658 CDS) were annotated as a result of work done directly on H.pylori genes. Finally, comparison of the two published genomes revealed a significant amount of size variation between corresponding (orthologous) CDS. Most of these size variations were due to natural polymorphisms, although other sources of variation were identified, such as pseudogenes, new genes potentially regulated by slipped strand mispairing mechanism, or frame-shifts. 113 of these differences were due to different start codon assignments, a common problem when constructing physical annotations. PMID- 12626711 TI - Histone deacetylase inhibitors stimulate mitochondrial HMG-CoA synthase gene expression via a promoter proximal Sp1 site. AB - The expression of mitochondrial HMG-CoA synthase in the colon has been correlated with the levels of butyrate present in this tissue. We report here that the effect of butyrate on mitochondrial HMG-CoA synthase gene expression is exerted in vivo at the transcriptional level, and that trichostatin A (TSA), a specific histone deacetylase inhibitor, also induces transcriptional activity and mRNA expression of the gene in human cell lines derived from colon carcinoma. Using chromatin immunoprecipitation assays, we show that histone deacetylase 1 (HDAC1) is associated with the endogenous mitochondrial HMG-CoA synthase promoter and that TSA induction correlates with hyperacetylation of H4 histone associated with the 5' flanking region of the gene. Overexpression of HDAC1 activity leads consistently to mitochondrial HMG-CoA synthase promoter hypoacetylation and reduces its transcriptional activity. The effect of butyrate and TSA maps to a single Sp1 site present in the proximal promoter of the gene, which is able to bind Sp1 and Sp3 proteins. Interestingly, the binding affinity of Sp1 and Sp3 proteins to the Sp1 site correlates with the TSA responsiveness of the promoter. Using a one-hybrid system (GAL4-Sp1 and GAL4-Sp3), we show that both proteins can mediate responsiveness to TSA in CaCo-2 cells employing distinct mechanisms. PMID- 12626713 TI - The ATM-related Tel1 protein of Saccharomyces cerevisiae controls a checkpoint response following phleomycin treatment. AB - MEC1 and TEL1 encode ATR- and ATM-related proteins in the budding yeast Saccharomyces cerevisiae, respectively. Phleomycin is an agent that catalyzes double-strand breaks in DNA. We show here that both Mec1 and Tel1 regulate the checkpoint response following phleomycin treatment. MEC1 is required for Rad53 phosphorylation and cell-cycle progression delay following phleomycin treatment in G1, S or G2/M phases. The tel1Delta mutation confers a defect in the checkpoint responses to phleomycin treatment in S phase. In addition, the tel1Delta mutation enhances the mec1 defect in activation of the phleomycin induced checkpoint pathway in S phase. In contrast, the tel1Delta mutation confers only a minor defect in the checkpoint responses in G1 phase and no apparent defect in G2/M phase. Methyl methanesulfonate (MMS) treatment also activates checkpoints, inducing Rad53 phosphorylation in S phase. MMS-induced Rad53 phosphorylation is not detected in mec1Delta mutants during S phase, but occurs in tel1Delta mutants similar to wild-type cells. Finally, Xrs2 is phosphorylated after phleomycin treatment in a TEL1-dependent manner during S phase, whereas no significant Xrs2 phosphorylation is detected after MMS treatment. Together, our results support a model in which Tel1 contributes to checkpoint control in response to phleomycin-induced DNA damage in S phase. PMID- 12626714 TI - RPA is an initiation factor for human chromosomal DNA replication. AB - The initiation of chromosomal DNA replication in human cell nuclei is not well understood because of its complexity. To allow investigation of this process on a molecular level, we have recently established a cell-free system that initiates chromosomal DNA replication in an origin-specific manner under cell cycle control in isolated human cell nuclei. We have now used fractionation and reconstitution experiments to functionally identify cellular factors present in a human cell extract that trigger initiation of chromosomal DNA replication in this system. Initial fractionation of a cytosolic extract indicates the presence of at least two independent and non-redundant initiation factors. We have purified one of these factors to homogeneity and identified it as the single-stranded DNA binding protein RPA. The prokaryotic single-stranded DNA binding protein SSB cannot substitute for RPA in the initiation of human chromosomal DNA replication. Antibodies specific for human RPA inhibit the initiation step of human chromosomal DNA replication in vitro. RPA is recruited to DNA replication foci and becomes phosphorylated concomitant with the initiation step in vitro. These data establish a direct functional role for RPA as an essential factor for the initiation of human chromosomal DNA replication. PMID- 12626715 TI - An NMR study on the interaction of Escherichia coli DinI with RecA-ssDNA complexes. AB - The SOS response, a set of cellular phenomena exhibited by eubacteria, is initiated by various causes that include DNA damage-induced replication arrest, and is positively regulated by the co- protease activity of RecA. Escherichia coli DinI, a LexA-regulated SOS gene product, shuts off the initiation of the SOS response when overexpressed in vivo. Biochemical and genetic studies indicated that DinI physically interacts with RecA to inhibit its co-protease activity. Using nuclear magnetic resonance (NMR) spectroscopy, we show that DinI tightly binds to the central region of RecA (between the N- and C-terminal domains) and that this interaction is enhanced upon the oligomerisation of RecA. On the other hand, DinI did not inhibit the interaction between 4mer single-stranded (ss)DNA and RecA- ATPgammaS, but had a slight effect on the structure of ssDNA-RecA ATPgammaS complexes involving 8mer and 12mer ssDNA. We hypothesise that prevention of repressor binding to the intermolecular cleft region of RecA protomers by DinI, with the possibility of a slight conformational change induced in the DinI-bound ssDNA-RecA-ATPgammaS complex, together function to inhibit the co-protease activity of RecA. PMID- 12626716 TI - Functional dissection of the zinc finger and flanking domains of the Yth1 cleavage/polyadenylation factor. AB - Yth1, a subunit of yeast Cleavage Polyadenylation Factor (CPF), contains five CCCH zinc fingers. Yth1 was previously shown to interact with pre-mRNA and with two CPF subunits, Brr5/Ysh1 and the polyadenylation-specific Fip1, and to act in both steps of mRNA 3' end processing. In the present study, we have identified new domains involved in each interaction and have analyzed the consequences of mutating these regions on Yth1 function in vivo and in vitro. We have found that the essential fourth zinc finger (ZF4) of Yth1 is critical for interaction with Fip1 and RNA, but not for cleavage, and a single point mutation in ZF4 impairs only polyadenylation. Deletion of the essential N-terminal region that includes the ZF1 or deletion of ZF4 weakened the interaction with Brr5 in vitro. In vitro assays showed that the N-terminus is necessary for both processing steps. Of particular importance, we find that the binding of Fip1 to Yth1 blocks the RNA Yth1 interaction, and that this inhibition requires the Yth1-interacting domain on Fip1. Our results suggest a role for Yth1 not only in the execution of cleavage and poly(A) addition, but also in the transition from one step to the other. PMID- 12626717 TI - Toucan: deciphering the cis-regulatory logic of coregulated genes. AB - TOUCAN is a Java application for the rapid discovery of significant cis regulatory elements from sets of coexpressed or coregulated genes. Biologists can automatically (i) retrieve genes and intergenic regions, (ii) identify putative regulatory regions, (iii) score sequences for known transcription factor binding sites, (iv) identify candidate motifs for unknown binding sites, and (v) detect those statistically over-represented sites that are characteristic for a gene set. Genes or intergenic regions are retrieved from Ensembl or EMBL, together with orthologs and supporting information. Orthologs are aligned and syntenic regions are selected as candidate regulatory regions. Putative sites for known transcription factors are detected using our MotifScanner, which scores position weight matrices using a probabilistic model. New motifs are detected using our MotifSampler based on Gibbs sampling. Binding sites characteristic for a gene set -and thus statistically over-represented with respect to a reference sequence set -are found using a binomial test. We have validated Toucan by analyzing muscle specific genes, liver-specific genes and E2F target genes; we have easily detected many known binding sites within intergenic DNA and identified new biologically plausible sites for known and unknown transcription factors. Software available at http://www.esat.kuleuven.ac. be/ approximately dna/BioI/Software.html. PMID- 12626718 TI - The affinity of different MBD proteins for a specific methylated locus depends on their intrinsic binding properties. AB - The methyl-CpG binding domain (MBD) family of proteins was defined based on sequence similarity in their DNA binding domains. In light of their high degree of conservation, it is of inherent interest to determine the genomic distribution of these proteins, and their associated co-repressor complexes. One potential determinant of specificity resides in differences in the intrinsic DNA binding properties of the various MBD proteins. In this report, we use a capillary electrophoretic mobility shift assay (CEMSA) with laser-induced fluorescence (LIF) and neutral capillaries to calculate MBD-DNA binding affinities. MBD proteins were assayed on pairs of methylated and unmethylated duplex oligos corresponding to the promoter regions of the BRCA1, MLH1, GSTP1 and p16(INK4a) genes, and binding affinities for each case were calculated by Scatchard analyses. With the exception of mammalian MBD3 and Xenopus MBD3 LF, all the MBD proteins showed higher affinity for methylated DNA (in the nanomolar range) than for unmethylated DNA (in the micromolar range). Significant differences between MBD proteins in the affinity for methylated DNA were observed, ranging within two orders of magnitude. By mutational analysis of MBD3 and using CEMSA, we demonstrate the critical role of specific residues within the MBD in conferring selectivity for methylated DNA. Interestingly, the binding affinity of specific MBD proteins for methylated DNA fragments from naturally occurring sequences are affected by local methyl-CpG spacing. PMID- 12626720 TI - ZCURVE: a new system for recognizing protein-coding genes in bacterial and archaeal genomes. AB - A new system, ZCURVE 1.0, for finding protein- coding genes in bacterial and archaeal genomes has been proposed. The current algorithm, which is based on the Z curve representation of the DNA sequences, lays stress on the global statistical features of protein-coding genes by taking the frequencies of bases at three codon positions into account. In ZCURVE 1.0, since only 33 parameters are used to characterize the coding sequences, it gives better consideration to both typical and atypical cases, whereas in Markov-model-based methods, e.g. Glimmer 2.02, thousands of parameters are trained, which may result in less adaptability. To compare the performance of the new system with that of Glimmer 2.02, both systems were run, respectively, for 18 genomes not annotated by the Glimmer system. Comparisons were also performed for predicting some function known genes by both systems. Consequently, the average accuracy of both systems is well matched; however, ZCURVE 1.0 has more accurate gene start prediction, lower additional prediction rate and higher accuracy for the prediction of horizontally transferred genes. It is shown that the joint applications of both systems greatly improve gene-finding results. For a typical genome, e.g. Escherichia coli, the system ZCURVE 1.0 takes approximately 2 min on a Pentium III 866 PC without any human intervention. The system ZCURVE 1.0 is freely available at: http://tubic. tju.edu.cn/Zcurve_B/. PMID- 12626719 TI - Glutamate at the phosphorylation site of response regulator CtrA provides essential activities without increasing DNA binding. AB - The essential response regulator CtrA controls the Caulobacter crescentus cell cycle and phosphorylated CtrA approximately P preferentially binds target DNA in vitro. The CtrA aspartate to glutamate (D51E) mutation mimics phosphorylated CtrA approximately P in vivo and rescues non-viable C.crescentus cells. However, we observe that the CtrA D51E and the unphosphorylated CtrA wild-type proteins have identical DNA affinities and produce identical DNase I protection footprints inside the C.crescentus replication origin. There fore, D51E promotes essential CtrA activities separate from increased DNA binding. Accordingly, we argue that CtrA protein recruitment to target DNA is not sufficient to regulate cell cycle progression. PMID- 12626722 TI - Primer-design for multiplexed genotyping. AB - Single-nucleotide polymorphism (SNP) analysis is a powerful tool for mapping and diagnosing disease-related alleles. Mutation analysis by polymerase-mediated single-base primer extension (minisequencing) can be massively parallelized using DNA microchips or flow cytometry with microspheres as solid support. By adding a unique oligonucleotide tag to the 5' end of the minisequencing primer and attaching the complementary antitag to the array or bead surface, the assay can be 'demultiplexed'. Such high-throughput scoring of SNPs requires a high level of primer multiplexing in order to analyze multiple loci in one assay, thus enabling inexpensive and fast polymorphism scoring. We present a computer program to automate the design process for the assay. Oligonucleotide primers for the reaction are automatically selected by the software, a unique DNA tag/antitag system is generated, and the pairing of primers and DNA tags is automatically done in a way to avoid any crossreactivity. We report results on a 45-plex genotyping assay, indicating that minisequencing can be adapted to be a powerful tool for high-throughput, massively parallel genotyping. The software is available to academic users on request. PMID- 12626721 TI - Micromolar concentrations of hydrogen peroxide induce oxidative DNA lesions more efficiently than millimolar concentrations in mammalian cells. AB - Reactive oxygen species produce oxidized bases, deoxyribose lesions and DNA strand breaks in mammalian cells. Previously, we demonstrated that aldehydic DNA lesions (ADLs) were induced in mammalian cells by 10 mM hydrogen peroxide (H2O2). Interestingly, a bimodal H2O2 dose-response relationship in cell toxicity has been reported for Escherichia coli deficient in DNA repair as well as Chinese hamster ovary (CHO) cells. Furthermore, it has been demonstrated that H2O2 causes single-strand breaks in purified DNA in the presence of iron and induces mitochondrial DNA damage in CHO cells with a biphasic dose-response curve. Here we show that H2O2 produces ADLs at concentrations as low as 0.06 mM in HeLa cells and that lower concentrations of H2O2 were much more efficient at inducing ADLs than higher concentrations. This dose-response curve is strikingly similar to that for cell killing effects in E.coli deficient in DNA repair exposed to H2O2. Interestingly, serial treatment of submillimolar levels of H2O2 induced a massive accumulation of ADLs. The toxicity arising from H2O2 determined by intracellular NAD(P)H in cells correlated well with the formation of ADLs. The addition of dipyridyl, an iron (II)-specific chelator, significantly protected against DNA damage and cell toxicity from submillimolar, but not millimolar, amounts of H2O2. These results suggest that ADLs induced by submillimolar levels of H2O2 may be due to a Fenton-type reaction between H2O2 and intracellular iron ions in mammalian cells. PMID- 12626724 TI - Nucleic acid capture assay, a new method for direct quantitation of nucleic acids. AB - Technologies allowing direct detection of specific RNA/DNA sequences occasionally serve as an alternative to amplification methods for gene expression studies. In these direct methods the hybridization of probes takes place in complex mixtures, thus specificity and sensitivity still limit the use of current technologies. To address these challenges, we developed a new technique called the nucleic acid capture assay, involving a direct multi-capture system. This approach combines a 3'-ethylene glycol scaffolding with the incorporation of 2'-methoxy deoxyribonucleotides in the capture sequences. In our design, all nucleotides other than those complementary to the target mRNA have been replaced by an inert linker, resulting in significant reductions in non-specific binding. We also provide a versatile method to detect the presence of captured targets by using specific labeled probes with alkaline phosphatase-conjugated anti-label antibodies. This direct, flexible and reliable technique for gene expression analysis is well suited for high-throughput screening and has potential for DNA microarray applications. PMID- 12626723 TI - Small amplified RNA-SAGE: an alternative approach to study transcriptome from limiting amount of mRNA. AB - Serial analysis of gene expression (SAGE) is a widely used and powerful technique to characterize and compare transcriptomes. Although several modifications have been proposed to the initial protocol with the aim of reducing the amount of starting material, unless additional PCR steps are added, the technique is still limited by the need for at least 1 micro g of total RNA. As extra PCR amplification might introduce representation biases, current SAGE protocols are not fully suitable for the study of small, microdissected tissue samples. We propose here an alternative method involving the linear amplification of small mRNA fragments containing the SAGE tags. The procedure allows preparation of libraries of over 100 000 tags from as few as 2500 cells. A satisfactory correlation was observed between a microSAGE library made from 5 microg of total thyroid RNA, and a library prepared from 50 ng of the same RNA preparation according to the present protocol. PMID- 12626685 TI - Bacteriophage T4 genome. AB - Phage T4 has provided countless contributions to the paradigms of genetics and biochemistry. Its complete genome sequence of 168,903 bp encodes about 300 gene products. T4 biology and its genomic sequence provide the best-understood model for modern functional genomics and proteomics. Variations on gene expression, including overlapping genes, internal translation initiation, spliced genes, translational bypassing, and RNA processing, alert us to the caveats of purely computational methods. The T4 transcriptional pattern reflects its dependence on the host RNA polymerase and the use of phage-encoded proteins that sequentially modify RNA polymerase; transcriptional activator proteins, a phage sigma factor, anti-sigma, and sigma decoy proteins also act to specify early, middle, and late promoter recognition. Posttranscriptional controls by T4 provide excellent systems for the study of RNA-dependent processes, particularly at the structural level. The redundancy of DNA replication and recombination systems of T4 reveals how phage and other genomes are stably replicated and repaired in different environments, providing insight into genome evolution and adaptations to new hosts and growth environments. Moreover, genomic sequence analysis has provided new insights into tail fiber variation, lysis, gene duplications, and membrane localization of proteins, while high-resolution structural determination of the "cell-puncturing device," combined with the three-dimensional image reconstruction of the baseplate, has revealed the mechanism of penetration during infection. Despite these advances, nearly 130 potential T4 genes remain uncharacterized. Current phage-sequencing initiatives are now revealing the similarities and differences among members of the T4 family, including those that infect bacteria other than Escherichia coli. T4 functional genomics will aid in the interpretation of these newly sequenced T4-related genomes and in broadening our understanding of the complex evolution and ecology of phages-the most abundant and among the most ancient biological entities on Earth. PMID- 12626725 TI - Novel amplification of DNA in a hairpin structure: towards a radical elimination of PCR errors from amplified DNA. AB - Errors introduced during PCR amplification set a selectivity limit for microsatellite analysis and molecular mutation detection methods since polymerase misincorporations invariably get confused with genuine mutations. Here we present hairpin-PCR, a new form of PCR that completely separates genuine mutations from polymerase misincorporations. Hairpin-PCR operates by converting a DNA sequence to a hairpin following ligation of oligonucleotide caps to DNA ends. We developed conditions that allow a DNA hairpin to be efficiently PCR-amplified so that, during DNA synthesis, the polymerase copies both DNA strands in a single pass. Consequently, when a misincorporation occurs it forms a mismatch following DNA amplification, and is distinguished from genuine mutations that remain fully matched. Error-free DNA can subsequently be isolated using one of many approaches, such as dHPLC or enzymatic depletion. We present feasibility for the main technical steps involved in this new strategy, conversion of a sequence to a hairpin that can be PCR-amplified from human genomic DNA, exponential amplification from picogram amounts, conversion of misincorporations to mismatches and separation of homoduplex from heteroduplex hairpins using dHPLC. The present hairpin-PCR opens up the possibility for a radical elimination of PCR errors from amplified DNA and a major improvement in mutation detection. PMID- 12626726 TI - Targeted gene modification in mismatch-repair-deficient embryonic stem cells by single-stranded DNA oligonucleotides. AB - Gene targeting through homologous recombination in murine embryonic stem (ES) cells is already strongly suppressed by DNA mismatch-repair (MMR)-dependent anti recombination when targeting construct and target locus differ at <1% of the nucleotide positions. We demonstrate that MMR activity also raises a strong impediment to gene modification mediated by small synthetic DNA oligonucleotide sequences. In the absence of the DNA MMR gene MSH2, synthetic single-stranded deoxyribo-oligonucleotides can be used to site-specifically modify the ES cell genome. We show that PCR-based procedures can be used to identify and clone modified cells. By this method we have substituted a single codon in the retinoblastoma gene. PMID- 12626727 TI - Adaptation of the Ras-recruitment system to the analysis of interactions between membrane-associated proteins. AB - Interactions of membrane-associated proteins play important roles in many cellular processes. The yeast two-hybrid assay is of limited utility for the analysis of such interactions, due to the need for soluble protein partners, whose interaction is assessed in the nucleus. The advent of the Ras-recruitment system (RRS) has enabled the study of membrane-associated proteins interacting with cytoplasmic proteins fused to Ras. Constitutive membrane association of the Ras fusion protein is expected to complement the growth defect of the yeast strain CDC25-2, assayed in the RRS, independent from the interaction with a membrane-bound partner. We describe the adaptation of the RRS to the analysis of interactions between two membrane-associated proteins using a model system. These results may facilitate the study of protein-protein interactions between membrane bound proteins and further increase the utility of the RRS. PMID- 12626729 TI - Analyzing partially randomized nucleic acid pools: straight dope on doping. AB - Partially randomized (doped) pools are important for optimizing activities initially isolated by selection-amplification or SELEX, and for locating nucleotides critical for function. Here we present a method for calculating the number of unique sequences in a pool, and the expected copy number of each unique sequence with a specified number of changes from the original sequence. Surprisingly, small differences in doping can have large consequences for the number of copies of sequences with certain numbers of changes from the original sequence. We demonstrate the effects of pool size, percentage doping, length of the random region and taking aliquots from the original pool on the exploration of sequence space in a doped reselection experiment. A web form is provided for customized calculations. PMID- 12626728 TI - Optimum conditions for selective isolation of genes from complex genomes by transformation-associated recombination cloning. AB - Transformation-associated recombination (TAR) cloning in yeast is used to isolate a desired chromosomal region or gene from a complex genome without construction of a genomic library. The technique involves homologous recombination during yeast spheroplast transformation between genomic DNA and a TAR vector containing short 5' and 3' gene-specific targeting hooks. Efficient gene capture requires a high yield of transformants, and we demonstrate here that the transformant yield increases approximately 10-fold when the genomic DNA is sheared to 100-200 kb before being presented to the spheroplasts. Here we determine the most effective concentration of genomic DNA, and also show that the targeted sequences recombine much more efficiently with the vector's targeting hooks when they are located at the ends of the genomic DNA fragment. We demonstrate that the yield of gene positive clones increases approximately 20-fold after endonuclease digestion of genomic DNA, which caused double strand breaks near the targeted sequences. These findings have led to a greatly improved protocol. PMID- 12626731 TI - Optical properties and ultrafast dynamics of metallic nanocrystals. AB - Noble metal particles have long fascinated scientists because of their intense color, which led to their application in stained glass windows as early as the Middle Ages. The recent resurrection of colloidal and cluster chemistry has brought about the strive for new materials that allow a bottoms-up approach of building improved and new devices with nanoparticles or artificial atoms. In this review, we discuss some of the properties of individual and some assembled metallic nanoparticles with a focus on their interaction with cw and pulsed laser light of different energies. The potential application of the plasmon resonance as sensors is discussed. PMID- 12626730 TI - Site-specific mutagenesis by triple helix-forming oligonucleotides containing a reactive nucleoside analog. AB - The specific recognition of homopurine-homo pyrimidine regions in duplex DNA by triplex-forming oligonucleotides (TFOs) provides an attractive strategy for genetic manipulation. Alkylation of nucleobases with functionalized TFOs would have the potential for site-directed mutagenesis. Recently, we demonstrated that a TFO bearing 2-amino-6-vinylpurine derivative, 1, achieves triplex-mediated reaction with high selectivity toward the cytosine of the G-C target site. In this report, we have investigated the use of this reagent to target mutations to a specific site in a shuttle vector plasmid, which replicates in mammalian cells. TFOs bearing 1 produced adducts at the complementary position of 1 and thereby introduced mutations at that site during replication/repair of the plasmid in mammalian cells. Reagents that produce covalent cytosine modifications are relatively rare. These TFOs enable the preparation of templates carrying targeted cytosine adducts for in vitro and in vivo studies. The ability to target mutations may prove useful as a tool for studying DNA repair, and as a technique for gene therapy and genetic engineering. PMID- 12626732 TI - Molecular aspects of halide ion hydration: the cluster approach. AB - This review provides a historical context for our understanding of the hydration shell surrounding halide ions and illustrates how the cluster systems can be used, in combination with theory, to elucidate the behavior of water molecules in direct contact with the anion. We discuss how vibrational predissociation spectroscopy, carried out with weakly bound argon atoms, has been employed to deduce the morphology of the small water networks attached to anions in the primary steps of hydration. We emphasize the importance of charge-transfer in the binary interaction, and discuss how this process affects the structures of the larger networks. Finally, we survey how the negatively charged water clusters (H2O)n(-) are providing a molecular-level perspective on how diffuse excess electrons interact with the water networks. PMID- 12626733 TI - Tunneling and optical spectroscopy of semiconductor nanocrystals. AB - The use of a combination of tunneling and optical spectroscopy to investigate the size and shape-dependent level structure and single-electron charging phenomena in semiconductor nanocrystals is reviewed. The artificial atom character of semiconductor nanocrystal quantum dots is manifested in both the discrete level structure and in the charging multiplicity of the single-electron tunneling data, revealing s and p atomic-like states. Such states can be directly imaged using scanning tunneling microscopy, providing the extent and symmetry of the envelope wavefunctions. A detailed description of the effect of the tunneling geometry on the single-electron tunneling spectra is presented. Correlation of the optical and tunneling data allows for the assignment of the level spectrum. The generality of this powerful combination is further demonstrated in the study of quantum rods that manifest the transition from zero-dimensional quantum dots to one-dimensional quantum wires. PMID- 12626734 TI - Theory of dipole-bound anions. AB - The problem of the binding of an excess electron to polar molecules and their clusters has long fascinated researchers. Although excess electrons bound to such species tend to be very extended spatially and to have little spatial overlap with the valence electrons of the neutral molecules, inclusion of electron correlation effects is essential for quantitatively describing the electron binding. The major electron correlation contribution may be viewed as a dispersion interaction between the excess electron and the electrons of the molecule or cluster. Recent work using a one-electron Drude model to describe excess electrons interacting with polar molecules is reviewed. PMID- 12626735 TI - STM control of chemical reaction: single-molecule synthesis. AB - The fascinating advances in single atom/molecule manipulation with a scanning tunneling microscope (STM) tip allow scientists to fabricate atomic-scale structures or to probe chemical and physical properties of matters at an atomic level. Owing to these advances, it has become possible for the basic chemical reaction steps, such as dissociation, diffusion, adsorption, readsorption, and bond-formation processes, to be performed by using the STM tip. Complete sequences of chemical reactions are able to induce at a single-molecule level. New molecules can be constructed from the basic molecular building blocks on a one-molecule-at-a-time basis by using a variety of STM manipulation schemes in a systematic step-by-step manner. These achievements open up entirely new opportunities in nanochemistry and nanochemical technology. In this review, various STM manipulation techniques useful in the single-molecule reaction process are reviewed, and their impact on the future of nanoscience and technology are discussed. PMID- 12626736 TI - Two-dimensional femtosecond spectroscopy. AB - The simplest two-dimensional (2D) spectra show how excitation with one (variable) frequency affects the spectrum at all other frequencies, thus revealing the molecular connections between transitions. Femtosecond 2D Fourier transform (2D FT) spectra are more flexible and share some of the remarkable properties of their conceptual parent, 2D FT nuclear magnetic resonance. When 2D FT spectra are experimentally separated into real absorptive and imaginary refractive parts, the time resolution and frequency resolution can both reach the uncertainty limit set for each resonance by the sample itself. Coherent four-level contributions to the signal provide new molecular phase information, such as relative signs of transition dipoles. The nonlinear response can be picked apart by selecting a single coherence pathway (e.g., specifying the relative signs of energy level difference frequencies during different time intervals as in the photon echo). Because molecules are frozen on the femtosecond timescale, femtosecond 2D FT experiments can separate a distribution of instantaneous molecular environments and intramolecular geometries as inhomogeneous broadening. This review provides an introduction to two-dimensional Fourier transform experiments exploiting second- and third-order vibrational and electronic nonlinearities. PMID- 12626737 TI - Transcriptional network controlled by the trithorax-group gene ash2 in Drosophila melanogaster. AB - The transcription factor absent, small, or homeotic discs 2 (ash2) gene is a member of the trithorax group of positive regulators of homeotic genes. Mutant alleles for ash2 are larvalpupal lethals and display imaginal disc and brain abnormalities. The allele used in this study is a true mutant for the trithorax function and lacks the longest transcript present in wild-type flies. In an attempt to identify gene targets of ash2, we have performed an expression analysis by using cDNA microarrays. Genes involved in cell cycle, cell proliferation, and cell adhesion are among these targets, and some of them are validated by functional and expression studies. Even though trithorax proteins act by modulating chromatin structure at particular chromosomal locations, evidence of physical aggregation of ash2-regulated genes has not been found. This work represents the first microarray analysis, to our knowledge, of a trithorax group gene. PMID- 12626738 TI - Transcription termination control of the S box system: direct measurement of S adenosylmethionine by the leader RNA. AB - Modulation of the structure of a leader RNA to control formation of an intrinsic termination signal is a common mechanism for regulation of gene expression in bacteria. Expression of the S box genes in Gram-positive organisms is induced in response to limitation for methionine. We previously postulated that methionine availability is monitored by binding of a regulatory factor to the leader RNA and suggested that methionine or S-adenosylmethionine (SAM) could serve as the metabolic signal. In this study, we show that efficient termination of the S box leader region by bacterial RNA polymerase depends on SAM but not on methionine or other related compounds. We also show that SAM directly binds to and induces a conformational change in the leader RNA. Both binding of SAM and SAM-directed transcription termination were blocked by leader mutations that cause constitutive expression in vivo. Overproduction of SAM synthetase in Bacillus subtilis resulted in delay in induction of S box gene expression in response to methionine starvation, consistent with the hypothesis that SAM is the molecular effector in vivo. These results indicate that SAM concentration is sensed directly by the nascent transcript in the absence of a trans-acting factor. PMID- 12626739 TI - Integrating regulatory motif discovery and genome-wide expression analysis. AB - We propose motif regressor for discovering sequence motifs upstream of genes that undergo expression changes in a given condition. The method combines the advantages of matrix-based motif finding and oligomer motif-expression regression analysis, resulting in high sensitivity and specificity. motif regressor is particularly effective in discovering expression-mediating motifs of medium to long width with multiple degenerate positions. When applied to Saccharomyces cerevisiae, motif regressor identified the ROX1 and YAP1 motifs from Rox1p and Yap1p overexpression experiments, respectively; predicted that Gcn4p may have increased activity in YAP1 deletion mutants; reported a group of motifs (including GCN4, PHO4, MET4, STRE, USR1, RAP1, M3A, and M3B) that may mediate the transcriptional response to amino acid starvation; and found all of the known cell-cycle regulation motifs from 18 expression microarrays over two cell cycles. PMID- 12626740 TI - Coadministration of HIV vaccine vectors with vaccinia viruses expressing IL-15 but not IL-2 induces long-lasting cellular immunity. AB - Vaccine efficacy is determined largely by cellular and humoral immunity as well as long-lasting immunological memory. IL-2 and IL-15 were evaluated in vaccinia vectors expressing HIV gp160 for the establishment of an effective vaccine strategy. Both IL-2 and IL-15 in the vaccinia vector induced strong and long lasting antibody-mediated immunity as well as a short-term cytotoxic T cell response against HIV gp120. In addition, IL-15 also supported robust CD8+ T cell mediated long-term immunity, whereas the CD8+ T cell-mediated immunity induced by IL-2 was short-lived. Moreover, we found that the cytokine milieu at the time of priming had surprisingly persistent effects on the character of the memory CD8 T cells long afterward with respect to their fate, functional activities, cytokine receptor expression, and antigen-independent proliferation. PMID- 12626741 TI - Protein pathway and complex clustering of correlated mRNA and protein expression analyses in Saccharomyces cerevisiae. AB - The mRNA and protein expression in Saccharomyces cerevisiae cultured in rich or minimal media was analyzed by oligonucleotide arrays and quantitative multidimensional protein identification technology. The overall correlation between mRNA and protein expression was weakly positive with a Spearman rank correlation coefficient of 0.45 for 678 loci. To place the data sets in a proper biological context, a clustering approach based on protein pathways and protein complexes was implemented. Protein expression levels were transcriptionally controlled for not only single loci but for entire protein pathways (e.g., Met, Arg, and Leu biosynthetic pathways). In contrast, the protein expression of loci in several protein complexes (e.g., SPT, COPI, and ribosome) was posttranscriptionally controlled. The coupling of the methods described provided insight into the biology of S. cerevisiae and a clustering strategy by which future studies should be based. PMID- 12626742 TI - Conditional tradeoffs between aging and organismal performance of Indy long-lived mutant flies. AB - Alterations that extend the life span of animals and yeast typically involve decreases in metabolic rate, growth, physical activity, and/or early-life fecundity. This negative correlation between life span and the ability to assimilate and process energy, to move, grow, and reproduce, raises questions about the potential utility of life span extension. Tradeoffs between early-life fitness and longevity are central to theories of the evolution of aging, which suggests there is necessarily a price to be paid for reducing the rate of aging. It is not yet clear whether life span can be extended without undesirable effects on metabolism and fecundity. Here, we report that the long-lived Indy mutation in Drosophila causes a decrease in the slope of the mortality curve consistent with a slowing in the rate of aging without a concomitant reduction in resting metabolic rate, flight velocity, or age-specific fecundity under normal rearing conditions. However, Indy mutants on a decreased-calorie diet have reduced fecundity, suggesting that a tradeoff between longevity and this aspect of performance is conditional, i.e., the tradeoff can occur in a stressful environment while being absent in a more favorable environment. These results provide evidence that there do exist mechanisms, albeit conditional, that can extend life span without significant reduction in fecundity, metabolic rate, or locomotion. PMID- 12626743 TI - Modifying the stereochemistry of an enzyme-catalyzed reaction by directed evolution. AB - Aldolases have potential as tools for the synthesis of stereochemically complex carbohydrates. Here, we show that directed evolution can be used to alter the stereochemical course of the reaction catalyzed by tagatose-1,6-bisphosphate aldolase. After three rounds of DNA shuffling and screening, the evolved aldolase showed an 80-fold improvement in k(cat)/K(m) toward the non-natural substrate fructose 1,6-bisphosphate, resulting in a 100-fold change in stereospecificity. (31)P NMR spectroscopy was used to show that, in the synthetic direction, the evolved aldolase catalyzes the formation of carbon-carbon bonds with unnatural diastereoselectivity, where the >99:<1 preference for the formation of tagatose 1,6-bisphosphate was switched to a 4:1 preference for the diastereoisomer, fructose 1,6-bisphosphate. This demonstration is of considerable significance to synthetic chemists requiring efficient syntheses of complex stereoisomeric products, such as carbohydrate mimetics. PMID- 12626744 TI - An approach to electrical modeling of single and multiple cells. AB - Previous theoretical approaches to understanding effects of electric fields on cells have used partial differential equations such as Laplace's equation and cell models with simple shapes. Here we describe a transport lattice method illustrated by a didactic multicellular system model with irregular shapes. Each elementary membrane region includes local models for passive membrane resistance and capacitance, nonlinear active sources of the resting potential, and a hysteretic model of electroporation. Field amplification through current or voltage concentration changes with frequency, exhibiting significant spatial heterogeneity until the microwave range is reached, where cellular structure becomes almost "electrically invisible." In the time domain, membrane electroporation exhibits significant heterogeneity but occurs mostly at invaginations and cell layers with tight junctions. Such results involve emergent behavior and emphasize the importance of using multicellular models for understanding tissue-level electric field effects in higher organisms. PMID- 12626746 TI - Interplay of the Arabidopsis nonhost resistance gene NHO1 with bacterial virulence. AB - It is poorly understood why a particular plant species is resistant to the vast majority of potential pathogens that infect other plant species, a phenomenon referred to as "nonhost" resistance. Here, we show that Arabidopsis NHO1, encoding a glycerol kinase, is required for resistance to and induced by Pseudomonas syringae isolates from bean and tobacco. NHO1 is also required for resistance to the fungal pathogen Botrytis cinerea, indicating that NHO1 is not limited to bacterial resistance. Strikingly, P. s. pv. tomato DC3000, an isolate fully virulent on Arabidopsis, actively suppressed the NHO1 expression. This suppression is abolished in coi1 plants, indicating that DC3000 required an intact jasmonic acid signaling pathway in the plant to suppress NHO1 expression. Constitutive overexpression of NHO1 led to enhanced resistance to this otherwise virulent bacterium. The presence of avrB in DC3000, which activates a cultivar specific "gene-for-gene" resistance in Arabidopsis, restored the induction of NHO1 expression. Thus, NHO1 is deployed for both general and specific resistance in Arabidopsis and targeted by the bacterium for parasitism. PMID- 12626745 TI - Small ubiquitin-like modifier conjugation regulates nuclear export of TEL, a putative tumor suppressor. AB - Posttranslational modification by small ubiquitin-like modifier (SUMO) conjugation regulates the subnuclear localization of several proteins; however, SUMO modification has not been directly linked to nuclear export. The ETS (E Twenty-Six) family member TEL (ETV6) is a transcriptional repressor that can inhibit Ras-dependent colony growth in soft agar and induce cellular aggregation of Ras-transformed cells. TEL is frequently disrupted by chromosomal translocations such as the t(12;21), which is associated with nearly one-fourth of pediatric B cell acute lymphoblastic leukemia. In the vast majority of t(12;21)-containing cases, the second allele of TEL is deleted, suggesting that inactivation of TEL contributes to the disease. Although TEL functions in the nucleus as a DNA-binding transcriptional repressor, it has also been detected in the cytoplasm. Here we demonstrate that TEL is actively exported from the nucleus in a leptomycin B-sensitive manner. TEL is posttranslationally modified by sumoylation at lysine 99 within a highly conserved domain (the "pointed" domain). Mutation of the sumo-acceptor lysine or mutations within the pointed domain that affect sumoylation impair nuclear export of TEL. Mutation of lysine 99 also results in an increase in TEL transcriptional repression, presumably because of decreased nuclear export. We propose that the ability of TEL to repress transcription and suppress growth is regulated by sumoylation and nuclear export. PMID- 12626748 TI - Pituitary tumor transforming gene-null male mice exhibit impaired pancreatic beta cell proliferation and diabetes. AB - The mammalian securin, pituitary tumor transforming gene (PTTG), regulates sister chromatid separation during mitosis. Mice or cell lines deficient in PTTG expression, however, are surprisingly viable. Here we show that PTTG disruption in mice (PTTG-/-) severely impairs glucose homeostasis leading to diabetes during late adulthood, especially in males associated with nonautoimmune insulinopenia and reversed alphabeta cell ratio. Islet beta cell mass in PTTG-/- mice was already diminished before development of frank diabetes and only increased minimally during growth. BrdUrd incorporation of islet cells in PTTG-null mice was approximately 65% lower (P < 0.005) than in the WT pancreas, whereas apoptosis rates were similar. PTTG-/- beta cells had pleiotropic nuclei, suggesting defects in cell division. The results indicated that securin is indispensable for normal pancreatic beta cell proliferation. PMID- 12626747 TI - An A-71C substitution in a green gene at the second position in the red/green visual-pigment gene array is associated with deutan color-vision deficiency. AB - We studied 247 Japanese males with congenital deutan color-vision deficiency and found that 37 subjects (15.0%) had a normal genotype of a single red gene followed by a green gene(s). Two of them had missense mutations in the green gene(s), but the other 35 subjects had no mutations in either the exons or their flanking introns. However, 32 of the 35 subjects, including all 8 subjects with pigment-color defect, a special category of deuteranomaly, had a nucleotide substitution, A-71C, in the promoter of a green gene at the second position in the red/green visual-pigment gene array. Although the -71C substitution was also present in color-normal Japanese males at a frequency of 24.3%, it was never at the second position but always found further downstream. The substitution was found in 19.4% of Chinese males and 7.7% of Thai males but rarely in Caucasians or African Americans. These results suggest that the A-71C substitution in the green gene at the second position is closely associated with deutan color-vision deficiency. In Japanese and presumably other Asian populations further downstream genes with -71C comprise a reservoir of the visual-pigment genes that cause deutan color-vision deficiency by unequal crossing over between the intergenic regions. PMID- 12626749 TI - Senescence-specific gene expression fingerprints reveal cell-type-dependent physical clustering of up-regulated chromosomal loci. AB - Replicative senescence is the state of irreversible proliferative arrest that occurs as a concomitant of progressive telomere shortening. By using cDNA microarrays and the gabriel system of computer programs to apply domain-specific and procedural knowledge for data analysis, we investigated global changes in gene transcription occurring during replicative senescence in human fibroblasts and mammary epithelial cells (HMECs). Here we report the identification of transcriptional "fingerprints" unique to senescence, the finding that gene expression perturbations during senescence differ greatly in fibroblasts and HMECs, and the discovery that despite the disparate nature of the chromosomal loci affected by senescence in fibroblasts and HMECs, the up-regulated loci in both types of cells show physical clustering. This clustering, which contrasts with the random distribution of genes down-regulated during senescence or up regulated during reversible proliferative arrest (i.e., quiescence), supports the view that replicative senescence is associated with alteration of chromatin structure. PMID- 12626751 TI - A Saccharomyces cerevisiae mutant strain defective in acetyl-CoA carboxylase arrests at the G2/M phase of the cell cycle. AB - To elucidate the essential functions of acetyl-CoA carboxylase (ACC1FAS3) in Saccharomyces cerevisiae, a temperature-sensitive mutant (acc1(ts)) was constructed. When the acc1(ts) cells were synchronized in G(1) phase with alpha factor at the permissive temperature of 24 degrees C and then released from the blockade and incubated at the restrictive temperature of 37 degrees C, 95% of the cell population became arrested at the G(2)M phase of the cell cycle despite the presence of fatty acids (C(14)-C(26)) in the medium. These cells developed large undivided nuclei, and the spindles of the arrested mutant cells were short. Shifting the G(2) arrested cells back to the permissive temperature resulted in a reversal of the cell-cycle arrest, with cells initiating mitosis. However, after 3 h of incubation at 37 degrees C, G(2) arrested mutant cells lost viability and displayed a uniquely altered nuclear envelope. Using [1-(14)C]acetate as a precursor for fatty acids synthesis, we identified the phospholipids and sphingolipids derived from acc1(ts) cells and wild-type cells at 24 degrees C and 37 degrees C, respectively. The levels of inositol-ceramides [IPC, MIPC, and M(IP)(2)C] and very long-chain fatty acids C(24) and C(26) declined sharply in the G(2)M arrested cells because of ACC inactivation. Shifting the acc1(ts) cells to 24 degrees C after 2 h of incubation at 37 degrees C resulted in reactivation of the ACC and elevation of the ceramides and very long-chain fatty acid syntheses with normal cell-cycle progression. In contrast, synthesis of wild-type inositol-ceramides, C(24) and C(26), fatty acids were elevated on incubation at 37 degrees C and declined when the cells shifted to the permissive temperature of 24 degrees C. PMID- 12626750 TI - Potent inhibition of scrapie prion replication in cultured cells by bis acridines. AB - Prion diseases are characterized by an accumulation of PrP(Sc), a misfolded isoform of the normal cellular prion protein, PrP(C). We previously reported the bioactivity of acridine-based compounds against PrP(Sc) replication in scrapie infected neuroblastoma cells and now report the improved potency of bis-acridine compounds. Bis-acridines are characterized by a dimeric motif, comprising two acridine heterocycles tethered by a linker. A library of bis-(6-chloro-2-methoxy acridin-9-yl) and bis-(7-chloro-2-methoxy-benzo[b][1,5]naphthyridin-10-yl) analogs was synthesized to explore the effect of structurally diverse linkers on PrP(Sc) replication in scrapie-infected neuroblastoma cells. Structure-activity analysis revealed that linker length and structure are important determinants for inhibition of prion replication in cultured scrapied cells. Three bis-acridine analogs, (6-chloro-2-methoxy-acridin-9-yl)-(3-[4-[3-(6-chloro-2-methoxy-acridin-9 ylamino)-propyl]-piperazin-1-yl]-propyl)-amine, N,N'-bis-(6-chloro-2-methoxy acridin-9-yl)-1,8-diamino-3,6-dioxaoctane, and (1-[[4-(6-chloro-2-methoxy-acridin 9-ylamino)-butyl]-[3-(6-chloro-2-methoxy-acridin-9-ylamino)-propyl]-carbamoyl] ethyl)-carbamic acid tert-butyl ester, showed half-maximal inhibition of PrP(Sc) formation at 40, 25, and 30 nM, respectively, and were not cytotoxic to uninfected neuroblastoma cells at concentrations of 500 nM. Our data suggest that bis-acridine analogs may provide a potent alternative to the acridine-based compound quinacrine, which is currently under clinical evaluation for the treatment of prion disease. PMID- 12626752 TI - Chiral tubule self-assembly from an achiral diynoic lipid. AB - Tubules possessing microm-scale chiral substructure self-assemble from an achiral isomer of the tubule-forming diynoic phosphatidylcholine, 1,2-bis(10,12 tricosadiynoyl)sn-glycero-3-phosphocholine [DC(8,9)PC], showing that molecular chirality is not essential for tubule formation. CD spectroscopy shows that these structures' helical sense of handedness instead originates in a spontaneous cooperative chiral symmetry-breaking process. We conclude that the chiral symmetry-breaking must originate in the unusual feature common to the chiral and achiral tubule-forming molecules, the diynes centered in their hydrocarbon tails. PMID- 12626753 TI - Liquid-liquid phase transition in compressed hydrogen from first-principles simulations. AB - The properties of compressed liquid hydrogen, the most abundant fluid in the universe, have been investigated by means of first-principles molecular dynamics at pressures between 75 and 175 GPa and temperatures closer to the freezing line than so far reported in shock-wave experiments. Evidence for a liquid-liquid transition between a molecular and a dissociated phase is provided. The transition is accompanied by a 6% increase in density and by metallization. This finding has important implications for our understanding of the interiors of giant planets and supports predictions of a quantum fluid state at low temperatures. PMID- 12626754 TI - Wnt-5A augments repopulating capacity and primitive hematopoietic development of human blood stem cells in vivo. AB - Human hematopoietic stem cells are defined by their ability to repopulate multiple hematopoietic lineages in the bone marrow of transplanted recipients and therefore are functionally distinct from hematopoietic progenitors detected in vitro. Although factors capable of regulating progenitors are well established, in vivo regulators of hematopoietic repopulating function are unknown. By using a member of the vertebrate Wnt family, Wnt-5A, the proliferation and differentiation of progenitors cocultured on stromal cells transduced with Wnt-5A or treated with Wnt-5A conditioned medium (CM) was unaffected. However, i.p. injection of Wnt-5A CM into mice engrafted with human repopulating cells increased multilineage reconstitution by >3-fold compared with controls. Furthermore, in vivo treatment of human repopulating cells with Wnt-5A CM produced a greater proportion of phenotypically primitive hematopoietic progeny that could be isolated and shown to possess enhanced progenitor function independent of continued Wnt-5A treatment. Our study demonstrates that Wnt-5A augments primitive hematopoietic development in vivo and represents an in vivo regulator of hematopoietic stem cell function in the human. Based on these findings, we suggest a potential role for activation of Wnt signaling in managing patients exhibiting poor hematopoietic recovery shortly after stem cell transplantation. PMID- 12626755 TI - Transposon telomeres are widely distributed in the Drosophila genus: TART elements in the virilis group. AB - Telomeres of most animals, plants, and unicellular eukaryotes are made up of tandem arrays of repeated DNA sequences produced by the enzyme telomerase. Drosophila melanogaster has an unusual variation on this theme; telomeres consist of tandem arrays of sequences produced by successive transpositions of two non LTR retrotransposons, HeT-A and TART. To explore the phylogenetic distribution of these variant telomeres, we have looked for TART homologues in a distantly related Drosophila species, virilis. We have found elements that, despite many differences in nucleotide sequence, retain significant amino acid similarity to TART from D. melanogaster. These D. virilis TART elements have features that characterize TART elements in D. melanogaster: (i) they are found in tandem arrays on chromosome ends, (ii) they are not found in euchromatin, and (iii) they produce both sense and antisense transcripts, with the antisense RNA being in excess. The D. virilis TART elements have one surprising feature: both of the ORFs contain long stretches of the trinucleotide repeat CAX, encoding polyglutamine (with a few interspersed histidines). These long polyglutamine stretches are conserved in the three D. virilis elements sequenced. They do not interrupt any domains of known function in the TART proteins and are not seen in TART proteins from other species. Comparison of the D. virilis and D. melanogaster telomeres suggests that the retrotransposon mechanism of telomere maintenance may have arisen before the separation of the genus Drosophila. PMID- 12626756 TI - The power of human brain magnetoencephalographic signals can be modulated up or down by changes in an attentive visual task. AB - This paper presents evidence indicating that the signals generated by neural responses to visual input can be either enhanced by increasing or suppressed by decreasing the area of the stimuli to which attention is directed. We used magnetoencephalography (MEG) to measure the frequency-tagged steady-state visual evoked responses of 11 subjects presented with two superimposed images flickering at different frequencies. Each image consisted of seven parallel bars of equal length; in any image, all bars were either red or green and either horizontal or vertical. At randomly chosen times during the experiments, any one of the three middle bars in either image transiently increased or decreased in width. Subjects were asked to attend to one image and ignore the other and to respond to changes in bar width in the attended image with a key press. In one condition, subject responses were required for changes in any of the three central bars of the attended image. We found that visual steady-state evoked responses to the attended image were enhanced relative to those evoked by the unattended image in this condition. In a second condition, subject responses were required for changes only in the middle bar. In this condition, the responses to the attended image were suppressed relative to those of the unattended image. These results may reflect relative differences in the synchronization and desynchronization of responding neuronal populations. PMID- 12626758 TI - Mutations that affect the ability of the vnd/NK-2 homeoprotein to regulate gene expression: transgenic alterations and tertiary structure. AB - The importance in downstream target regulation of tertiary structure and DNA binding specificity of the protein encoded by the vndNK-2 homeobox gene is analyzed. The ectopic expression patterns of WT and four mutant vndNK-2 genes are analyzed together with expression of two downstream target genes, ind and msh, which are down-regulated by vndNK-2. Three mutants are deletions of conserved regions (i.e., tinman motif, acidic motif, and NK-2 box), and the fourth, Y54M vndNK-2, corresponds to a single amino acid residue replacement in the homeodomain. Of the four ectopically expressed mutant genes examined, only the Y54M mutation inactivates the ability of the vndNK-2 homeodomain protein to repress ind and msh. The acidic motif deletion mutant slightly reduced the ability of the protein to repress ind and msh. By contrast, both tinman and NK-2 box deletion mutants behaved as functional vndNK-2 genes in their ability to repress ind and msh. The NMR-determined tertiary structures of the Y54M vndNK-2 homeodomain, both free and bound to DNA, are compared with the WT analog. The only structural difference observed for the mutant homeodomain is in the complex with DNA and involved closer interaction of the methionine-54 with A2, rather than with C3 of the (-) strand of the DNA. This subtle change in the homeodomain DNA complex resulted in modifications of binding affinities to DNA. These changes resulting from a single amino acid residue replacement constitute the molecular basis for the phenotypic alterations observed on ectopic expression of the Y54M vndNK-2 gene during embryogenesis. PMID- 12626757 TI - Mapping Wnt/beta-catenin signaling during mouse development and in colorectal tumors. AB - Wntbeta-catenin signaling plays key roles in several developmental and pathological processes. Domains of Wnt expression have been extensively investigated in the mouse, but the tissues receiving the signal remain largely unidentified. To define which cells respond to activated beta-catenin during mammalian development, we generated the beta-catenin-activated transgene driving expression of nuclear beta-galactosidase reporter (BAT-gal) transgenic mice, expressing the lacZ gene under the control of beta-cateninT cell factor responsive elements. Reporter gene activity is found in known organizing centers, such as the midhindbrain border and the limb apical ectodermal ridge. Moreover, BAT-gal expression identifies novel sites of Wnt signaling, like notochord, endothelia, and areas of the adult brain, revealing an unsuspected dynamic pattern of beta-catenin transcriptional activity. Expression of the transgene was analyzed in mutant backgrounds. In lipoprotein receptor-related protein 6-null homozygous mice, which lack a Wnt coreceptor, BAT-gal staining is absent in mutant tissues, indicating that BAT-gal mice are bona fide in vivo indicators of Wntbeta-catenin signaling. Analyses of BAT-gal expression in the adenomatous polyposis coli (multiple intestinal neoplasia+) background revealed betacatenin transcriptional activity in intestinal adenomas but surprisingly not in normal crypt cells. In summary, BAT-gal mice unveil the entire complexity of Wntbeta catenin signaling in mammals and have broad application potentials for the identification of Wnt-responsive cell populations in development and disease. PMID- 12626760 TI - The impact of the certainty context on the process of choice. AB - In this study we examine how the introduction of a reference lottery with nonrandom outcomes alters the way in which choices among pairs of lotteries are made, even if it does not alter the choices. We use different domains (some of the lotteries produce gains, other losses) and different contexts (one member of the pair, the reference lottery, may be either risky or certain). In our experiment, the change from gain to loss domain affects choices: subjects are risk averse in the gain domain, but not in the loss domain. On the contrary, the context effect of the certain lottery does not affect choices. However, the introduction of the certainty reference lottery affects two behavioral variables, response time and brain activation, in a dramatic way. This result suggests that the certainty lottery promotes a different process through which preferences are revealed, even if the differences among lotteries may not be large enough to induce different choices. PMID- 12626759 TI - Gene-environment interaction modulated by allelic heterogeneity in inflammatory diseases. AB - CARD15 is a major susceptibility gene for a frequent multifactorial chronic inflammatory bowel disorder, Crohn disease (CD). By using NF-kappaB activation assays, the cytosolic CARD15 was shown to efficiently detect bacterial peptidoglycan (PGN), reminiscent of the PGN recognition protein surveillance mechanism in Drosophila. The 3 CD-associated variants and 13 additional variants carried by CD patients demonstrated impaired PGN-dependent response revealing null, hypomorphic, or dominant-negative properties. Quantitative parametrization of this response, computed from the patients' CARD15 genotypes, was predictive of several variable CD manifestations. In contrast, CARD15 alleles associated with Blau's syndrome promoted PGN-independent NF-kappaB activation, an observation that accounts for the minimal microbial input in the etiology of this dominant, monogenic inflammatory disorder affecting solely aseptic sites. PMID- 12626761 TI - Melanoma inhibitor of apoptosis protein (ML-IAP) is a target for immune-mediated tumor destruction. AB - The identification of antigens associated with tumor destruction is a major goal of cancer immunology. Vaccination with irradiated tumor cells engineered to secrete granulocyte-macrophage colony stimulating factor generates potent, specific, and long-lasting antitumor immunity through improved tumor antigen presentation by dendritic cells and macrophages. A phase I clinical trial of this immunization strategy in patients with disseminated melanoma revealed the consistent induction in distant metastases of dense T and B cell infiltrates that effectuated substantial tumor necrosis and fibrosis. To delineate the target antigens of this vaccine-stimulated tumor destruction, we screened a melanoma cDNA expression library with postimmunization sera from a long-term responding patient (K030). High-titer IgG antibodies recognized melanoma inhibitor of apoptosis protein (ML-IAP), a caspase antagonist containing a single baculoviral IAP repeat and a COOH-terminal RING domain. Although K030 harbored antibodies to ML-IAP at the time of study entry, multiple courses of vaccination over 4 years increased antibody titers and elicited isotype switching. Moreover, lymphocyte infiltrates in necrotic metastases included CD4+ and CD8+ T cells specific for ML IAP, as revealed by proliferation, tetramer, enzyme-linked immunospot, and cytotoxicity analysis. Whereas melanoma cells in densely infiltrated lesions showed strong ML-IAP expression by immunohistochemistry, lethal disease progression was associated with the loss of ML-IAP staining and the absence of lymphocyte infiltrates. These findings demonstrate that ML-IAP can serve as a target for immune-mediated tumor destruction, but that antigen-loss variants can accomplish immune escape. PMID- 12626762 TI - Therapeutic photobiomodulation for methanol-induced retinal toxicity. AB - Methanol intoxication produces toxic injury to the retina and optic nerve, resulting in blindness. The toxic metabolite in methanol intoxication is formic acid, a mitochondrial toxin known to inhibit the essential mitochondrial enzyme, cytochrome oxidase. Photobiomodulation by red to near-IR radiation has been demonstrated to enhance mitochondrial activity and promote cell survival in vitro by stimulation of cytochrome oxidase activity. The present studies were undertaken to test the hypothesis that exposure to monochromatic red radiation from light-emitting diode (LED) arrays would protect the retina against the toxic actions of methanol-derived formic acid in a rodent model of methanol toxicity. Using the electroretinogram as a sensitive indicator of retinal function, we demonstrated that three brief (2 min, 24 s) 670-nm LED treatments (4 J/cm(2)), delivered at 5, 25, and 50 h of methanol intoxication, attenuated the retinotoxic effects of methanol-derived formate. Our studies document a significant recovery of rod- and cone-mediated function in LED-treated, methanol-intoxicated rats. We further show that LED treatment protected the retina from the histopathologic changes induced by methanol-derived formate. These findings provide a link between the actions of monochromatic red to near-IR light on mitochondrial oxidative metabolism in vitro and retinoprotection in vivo. They also suggest that photobiomodulation may enhance recovery from retinal injury and other ocular diseases in which mitochondrial dysfunction is postulated to play a role. PMID- 12626763 TI - A binding motif for Siah ubiquitin ligase. AB - The Drosophila SINA (seven in absentia) protein and its mammalian orthologs (Siah, seven in absentia homolog) are RING domain proteins that function in E3 ubiquitin ligase complexes and facilitate ubiquitination and degradation of a wide range of cellular proteins, including beta-catenin. Despite these diverse targets, the means by which SINASiah recognize substrates or binding proteins has remained unknown. Here we identify a peptide motif (RPVAxVxPxxR) that mediates the interaction of Siah protein with a range of protein partners. Sequence alignment and mutagenesis scanning revealed residues that are important to this interaction. This consensus sequence correctly predicted a high-affinity interaction with a peptide from the cytoskeletal protein plectin-1 (residues 95 117). The unusually high-affinity binding obtained with a 23-residue peptide (K(Dapp) = 29 nM with SINA) suggests that it may serve as a useful dominant negative reagent for SINASiah proteins. PMID- 12626764 TI - Computational and experimental analysis of DNA shuffling. AB - We describe a computational model of DNA shuffling based on the thermodynamics and kinetics of this process. The model independently tracks a representative ensemble of DNA molecules and records their states at every stage of a shuffling reaction. These data can subsequently be analyzed to yield information on any relevant metric, including reassembly efficiency, crossover number, type and distribution, and DNA sequence length distributions. The predictive ability of the model was validated by comparison to three independent sets of experimental data, and analysis of the simulation results led to several unique insights into the DNA shuffling process. We examine a tradeoff between crossover frequency and reassembly efficiency and illustrate the effects of experimental parameters on this relationship. Furthermore, we discuss conditions that promote the formation of useless "junk" DNA sequences or multimeric sequences containing multiple copies of the reassembled product. This model will therefore aid in the design of optimal shuffling reaction conditions. PMID- 12626765 TI - Persistence and brain circuitry. AB - The orbitofrontal and adjacent medial prefrontal cortex may play an important role in normal social functioning and affect modulation. Recent anatomical studies of this area of the prefrontal cortex have demonstrated a striking correspondence of fine-grained architectonic partitioning schemes in humans and nonhuman primates. This finding allows neurophysiological recording and anatomical connectivity data in animals to be considered together with functional imaging data and lesion studies in humans. In a functional MRI study, we show that individual differences in Persistence, a dimensional trait assessed with a seven-factor personality model, may be linked to specific areas in the lateral orbital and medial prefrontal cortex and the ventral striatum. These areas are part of an anatomical circuit that has been defined in nonhuman primates and has been implicated in functions related to behavioral persistence. These findings represent a fresh approach to linking normal individual differences in personality and behavior to specific neuronal structures and subsystems. PMID- 12626767 TI - Recent developments in our understanding of the physiological role of PP-fold peptide receptor subtypes. AB - The three peptides pancreatic polypeptide (PP), peptide YY (PYY), and neuropeptide Y (NPY) share a similar structure known as the PP-fold. There are four known human G-protein coupled receptors for the PP-fold peptides, namely Y1, Y2, Y4, and Y5, each of them being able to bind at least two of the three endogenous ligands. All three peptides are found in the circulation acting as hormones. Although NPY is only released from neurons, PYY and PP are primarily found in endocrine cells in the gut, where they exert such effects as inhibition of gall bladder secretion, gut motility, and pancreatic secretion. However, when PYY is administered in an experimental setting to animals, cloned receptors, or tissue preparations, it can mimic the effects of NPY in essentially all studies, making it difficult to study the effects of PP-fold peptides and to delineate what receptor and peptide accounts for a particular effect. Initial studies with transgenic animals confirmed the well-established action of NPY on metabolism, food-intake, vascular systems, memory, mood, neuronal excitability, and reproduction. More recently, using transgenic techniques and novel antagonists for the Y1, Y2, and Y5 receptors, NPY has been found to be a key player in the regulation of ethanol consumption and neuronal development. PMID- 12626768 TI - From mouse to man: redefining the role of insulin-like growth factor-I in the acquisition of bone mass. AB - The insulin-like growth factor system (IGF) has been linked to the process of bone acquisition through epidemiologic analyses of large cohorts and in vitro studies of bone cells. But the exact relationship between expression of IGF-I in bone and skeletal homeostasis or pathologic conditions, such as osteoporosis, remains poorly defined. Recent advances in genomic engineering have resulted in the development of better in vivo models to test the role of IGF-I during development and maintenance of the adult skeleton. It is now established that skeletal expression of IGF-I is critical for differentiative bone cell function. It may also be essential for the full anabolic effects of parathyroid hormone on trabecular bone and for some component of biomineralization. Evidence from conditional mutagenesis studies suggests that serum IGF-I may represent more than a storage depot or permissive factor during the final phase of skeletal acquisition. This work re-examines the original tenets of the "somatomedin hypothesis" in light of these newer mouse models and their remarkable skeletal phenotypes. The implications are far reaching and suggest that newer approaches for manipulating the IGF regulatory system may one day be useful as therapeutic adjuncts for the treatment of osteoporosis. PMID- 12626769 TI - Astrocytes and brain function: implications for reproduction. AB - Recent evidence suggests that astrocytes have important neuroregulatory functions in addition to their classic functions of support and segregation of neurons. These newly revealed functions include regulation of neuron communication, neurosecretion, and synaptic plasticity. Although these actions occur throughout the brain, this review will focus on astrocyte-neuron interactions in the hypothalamus, particularly with respect to their potential contribution to the regulation of gonadotropin-releasing hormone (GnRH) secretion and reproduction. Hypothalamic astrocytes have been documented to release a variety of neuroactive factors, including transforming growth factors-alpha and -beta, insulin-like growth factor-1, prostaglandin E2, and the neurosteroid, 3 alpha-hydroxy-5 alpha pregnane-20-one. Each of these factors has been shown to stimulate GnRH release, and receptors for each factor have been documented on GnRH neurons. Astrocytes have also been implicated in the regulation of synaptic plasticity in key areas of the hypothalamus that control GnRH release, an effect achieved by extension and retraction of glial processes (i.e., glial ensheathment). Through this mechanism, the number of synapses on GnRH neurons and GnRH regulatory neurons can potentially be modulated, thereby influencing the activation state of GnRH neurons. The steroid hormone 17beta-estradiol, which triggers the GnRH and luteinizing hormone surge, has been shown to induce the astrocyte-regulated changes in hypothalamic synaptic plasticity, as well as enhance formation and release of the astrocyte neuroactive factors, thereby providing another potential mechanistic layer for astrocyte regulation of GnRH release. As a whole, these studies provide new insights into the diversity of astrocytes and their potential role in reproductive neuroendocrine function. PMID- 12626766 TI - Proteasome-dependent, ubiquitin-independent degradation of the Rb family of tumor suppressors by the human cytomegalovirus pp71 protein. AB - Most of the substrates degraded by the proteasome are marked with polyubiquitin chains. However, there are a limited number of examples of nonubiquitinated proteins that are degraded by the proteasome. Here, we describe the degradation of the retinoblastoma family of tumor suppressor proteins by the proteasome in the absence of polyubiquitination. The retinoblastoma protein (p105), p107, and p130 are each targeted for degradation by the pp71 protein, which is encoded by the UL82 gene of human cytomegalovirus. It functions to direct their degradation in the absence of other viral proteins. While the pp71-mediated degradation of the retinoblastoma family of proteins requires proteasome function, it occurs without the attachment of ubiquitin to the substrates and in the absence of a functioning ubiquitin-conjugation system. PMID- 12626770 TI - Expression of the prodynorphin gene after experimental brain injury and its role in behavioral dysfunction. AB - Traumatic brain injury (TBI) causes excess release of neurotransmitters, such as glutamate, and increases intracellular calcium levels. Elevated levels of calcium, and perhaps other intracellular second messengers, as a result of TBI can alter the expression of many genes. The protein products of some of these genes may be signals for TBI-associated memory dysfunction. Therefore, identification of genes whose expression is altered after TBI in the hippocampus, a structure in the medial temporal lobe that plays a critical role in memory formation and storage, and elucidation of the role(s) of their protein products may shed light on the molecular mechanisms underlying TBI-elicited memory dysfunction. The prodynorphin gene is expressed in hippocampal granule cells, and its expression has been reported to be enhanced as a result of elevated intracellular calcium. The prodynorphin protein is proteolytically cleaved to generate multiple dynorphin peptides, which can modulate neurotransmitter release through the activation of presynaptic kappa opioid receptors. In this study, we report that 1) TBI transiently increases prodynorphin mRNA in the hippocampus, 2) dynorphin peptide immunoreactivity is enhanced for up to 24 hr after TBI and 3) intracerebroventricular infusion of the kappa receptor antagonist nor binaltorphimine (nor-BNI) impairs subsequent performance in a spatial memory task. These results suggest that dynorphin action may serve a beneficial role after TBI. PMID- 12626771 TI - Tumor-derived cytokines dysregulate macrophage interferon-gamma responsiveness and interferon regulatory factor-8 expression. AB - Tumors can evade immune responses through suppressor signals that dysregulate host effector cell function. In this study we demonstrate that tumor-derived suppressor molecules impede host antitumor immune activity through dysregulation of multiple macrophage (Mphi) pathways, including suppressed production of cytotoxic and immunostimulatory agents and impaired expression of the interferon regulatory factor-8 (IRF-8) protein, a critical transducer of interferon-gamma mediated activation pathways. The tumor-derived immunosuppressive cytokines interleukin-10 and transforming growth factor-beta(1) constrain IRF-8 production by normal Mphis, regardless of priming, and IRF-8 is also dysregulated in primary Mphis from tumor-burdened hosts. Collectively, these data describe a new mechanism by which tumors disrupt immune function and suggest that abrogation of tumor-derived immunoregulatory factors in situ can restore immune function and enhance antitumor efficacy. PMID- 12626772 TI - Nulliparous CCAAT/enhancer binding proteindelta (C/EBPdelta) knockout mice exhibit mammary gland ductal hyperlasia. AB - CCAAT/Enhancer binding proteins (C/EBPs) are a family of nuclear proteins that function in the control of cell growth, death, and differentiation. We previously reported that C/EBPdelta plays a key role in mammary epithelial cell G(0) growth arrest. In this report, we investigated the role of C/EBPdelta in mammary gland development and function using female mice homozygous for a targeted deletion of C/EBPdelta (C/EBPdelta -/-). C/EBPdelta -/- females develop normally and exhibit normal reproductive and lactational performance. Adult nulliparous C/EBPdelta -/- females, however, exhibit mammary epithelial cell growth control defects. The mean number of mammary ductal branches is significantly higher in adult nulliparous C/EBPdelta -/- females compared with C/EBPdelta +/+ (wild-type control) females (66.8 +/- 5.2 vs 42.9 +/- 6.3 branch points/field, P < 0.01). In addition, the mean total mammary gland cellular volume occupied by epithelium is significantly higher in adult nulliparous C/EBPdelta -/- females compared with C/EBPdelta +/+ controls (29.0 +/- 1.4 vs 20.4 +/- 1.3, P < 0.001). Our results showed that the BrdU labeling index was significantly higher in mammary epithelial cells from nulliparous C/EBPdelta -/- females compared with C/EBPdelta +/+ controls during the proestrus/estrus (4.55 +/- 0.70 vs 2.14 +/- 0.43, P < 0.01) and metestrus/diestrus (6.92 +/- 0.75 vs 3.98 +/- 0.43 P < 0.01) phases of the estrus cycle. In contrast, the percentage of mammary epithelial cells undergoing apoptosis during both phases of the estrus cycle did not differ between C/EBPdelta -/- and C/EBPdelta +/+ females. The increased epithelial cell content and proliferative capacity was restricted to the nulliparous C/EBPdelta /- females as no differences in mammary gland morphology, ductal branching or total epithelial content were observed between multiparous C/EBPdelta -/- and C/EBPdelta +/+ females. These results demonstrate that C/EBPdelta plays a novel role in mammary epithelial cell growth control that appears to be restricted to the nulliparous mammary gland. PMID- 12626773 TI - Association of metallothionein expression and lack of apoptosis with progression of carcinogenesis in Barrett's esophagus. AB - Barrett's esophagus is the transformation of normal esophageal squamous epithelium to specialized intestinal metaplasia (SIM). Among the Barrett's specialized cells, those that can develop protective mechanisms against apoptosis may have potential to become malignant. Studies have shown that overexpression of metallothionein (MT), low molecular protein that protects cells from apoptotic stimuli, appears to be associated with more advanced, highly malignant tumors. We thus investigated the relationship between MT expression and apoptosis in different stages of Barrett's carcinogenesis. Terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling and immunohistochemical dual-staining assay were performed in human biopsy samples of normal, SIM, dysplasia, and adenocarcinoma. Apoptotic index and MT expression were quantified by using an image system to analyze the converted digital data. A negative correlation between MT expression and apoptotic index was found. MT expression was significantly increased along with the histologic progression towards adenocarcinoma. This study thus suggests that MT may contribute to cytoprotection, thereby inhibiting apoptosis and leading to carcinogenesis of Barrett's esophageal cells. PMID- 12626774 TI - Discovery of water-soluble anticancer agents (edotides) from a vegetable found in Benin City, Nigeria. AB - Cancer claims the lives of more than six million people each year in the world. About 1,268,000 new cancer cases, and 553,400 deaths were reported in the United States in 2001. Current treatment approaches have yielded significant progress in the fight against cancer, but the incidence of developing certain types of cancer continues to rise. This is especially true in the African-American communities. African Americans are about 33% more likely to die of cancer than are whites and more than twice likely to die of cancer as are Asian-Islander, American-Indians, and Hispanics. This increase coupled with the harsh side effects of some of the cancer chemotherapies have led to the search for more natural biological products, especially those derived from plant products, currently known as herbal medicine. There is a need for a continued search for novel natural products that may be used as cancer chemopreventive and/or chemotherapeutic agents. The objective of this study was to evaluate the effect(s) of a novel water-soluble leaf extract of Vernonia amygdalina (VA) on human breast cancer cell DNA synthesis. MCF-7 cell line, considered a suitable model, was used in this study. Treatment of cells with physiologically relevant concentrations of water-soluble VA extract potently inhibited DNA synthesis in a concentration-dependent fashion both in the absence and presence of serum. Fractions of VA extract separated using preparative reverse-phase chromatography also inhibited DNA synthesis (P < 0.005). These results suggest that VA vegetable, if incorporated in the diet, may prevent or delay the on-set of breast cancer. PMID- 12626775 TI - Effects of docosahexaenoic acid on vascular pathology and reactivity in hypertension. AB - Previous studies have shown that docosahexaenoic acid (DHA) has an antihypertensive effect in spontaneously hypertensive rats (SHR). To investigate possible mechanisms for this effect, vascular pathology and reactivity were determined in SHR treated with dietary DHA. SHR (7 weeks) were fed a purified diet with either a combination of corn/soybean oils or a DHA-enriched oil for 6 weeks. Histological evaluation of heart tissue, aorta, coronary, and renal arteries was performed. Vascular responses were determined in isolated aortic rings. Contractile responses to agonists, including norepinephrine (10(-9) to 10( 4) M), potassium chloride (5-55 mM), and angiotensin II (5 x 10(-7) M) were assessed. Vasorelaxant responses to acetylcholine (10(-9) to 10 (-4) M), sodium nitroprusside (10(-9) to 10(-6) M), papaverine (10(-5) to 10(-4) M), and methoxyverapamil (D600, 1-100 microM) were determined. DHA-fed SHR had significantly reduced blood pressure (P < 0.001) and vascular wall thicknesses in the coronary, thoracic, and abdominal aorta compared with controls (P < 0.05) Contractile responses to agonists mediated by receptor stimulation and potassium depolarization were not altered in DHA-fed SHR. Endothelial-dependent relaxations to acetylcholine were not altered which suggests endothelial-derived nitric oxide production/release is not affected by dietary DHA. Other mechanisms of vascular relaxation, including intracellular cyclic nucleotides, cGMP, and cAMP were not altered by dietary DHA because aortic relaxant responses to sodium nitroprusside and papaverine were similar in control and DHA-fed SHR. No significant differences were seen in relaxant responses to the calcium channel blocker, D600, or contractile responses to norepinephrine in the absence of extracellular calcium. These results suggest that dietary DHA does not affect mechanisms related to extracellular calcium channels or intracellular calcium mobilization. Moreover, the contractile and vasorelaxant responses are not differentially altered with dietary DHA in this in vivo SHR model. The findings demonstrate that dietary DHA reduces systolic blood pressure and vascular wall thickness in SHR. This may contribute to decrease arterial stiffness and pulse pressure, in addition to the antihypertensive properties of DHA. The antihypertensive properties of DHA are not related to alterations in vascular responses. PMID- 12626776 TI - Inhibitory effects of progesterone on plasma membrane fluidity and tumorigenic potential of ovarian epithelial cancer cells. AB - The lethality of common (surface) epithelial ovarian cancer is contingent on its metastatic capacity. Dissemination of the neoplasia throughout the abdominal cavity has been associated with secretion of proteolytic enzymes from vesicles shed by ovarian cancer cells. We report that the lipophilic steroid hormone progesterone decreases the fluid dynamics of plasma membranes of human SKOV-3 adenocarcinoma cells. The decrease in membrane fluidity was related to an inhibition in vitro of exocytotic vesicle release, cellular invasiveness into Matrigel, and colony formation in three-dimensional collagen matrix. Tumorigenesis was suppressed by progesterone in immunocompromised nude mice inoculated intraperitoneally with SKOV-3 cells. Progestins could therefore be of benefit in the prevention and(or) treatment of early-stage ovarian carcinomatosis. PMID- 12626777 TI - Enhanced intracellular calcium promotes metabolic and secretory disturbances in rat gastric mucosa during ethanol-induced gastritis. AB - Changes in the Ca(2+) homeostasis have been implicated in cell injury and death. However, Ca(2+) participation in ethanol-induced chronic gastric mucosal injury has not been elucidated. We have developed a model of ethanol-induced chronic gastric injury in rats, characterized by marked alterations in plasma membranes from gastric mucosa and a compensatory cell proliferation, which follows ethanol withdrawal. Therefore, the present study explored the possible role of intracellular Ca(2+) in the oxidative metabolism and in acid secretion in this experimental model. Glucose oxidation was greatly enhanced in the injured mucosa, as evaluated by CO(2) production by isolated mucosal preparations incubated with (14)C-radiolabeled glucose in different carbons. Oxygen consumption and acid secretion (aminopyrine accumulation) were also stimulated. A predominating secretory status was morphologically identified by electron microscopy in oxyntic cells of gastric mucosa from ethanol-treated rats. A coupling between secretory and metabolic effects induced by ethanol (demonstrated by an inhibitory effect of omeprazole in both parameters) was found. These ethanol-induced effects were also inhibited by addition of Ca(2+) chelators to isolated gastric mucosa samples. Lanthanum, a Ca(2+) channel blocker, inhibited ethanol-promoted increase of oxidative metabolism. In addition, a stimulated Ca(2+) uptake by mucosal minces and increased in vivo Ca(2+) levels in cytosolic and mitochondrial fractions, were also noticed. Enhanced glucose and oxygen consumptions were associated with higher ATP and NADP+ availability, whereas cytosolic NAD/NADH ratio (assessed by mucosal levels of lactate and pyruvate) was not significantly modified by the chronic ethanol administration. In conclusion, changes in Ca(2+) homeostasis, probably mainly due to increased extracellular Ca(2+) uptake, could mediate secretory and metabolic alterations found in the gastric mucosa from rats chronically treated with ethanol. PMID- 12626779 TI - Introduction: All Ireland Fatigue Coalition. AB - The first professional meeting and educational symposium of the All Ireland Fatigue Coalition (AIFC) convened in Dublin in September 2002, with an attendance of 350 health professionals who work in cancer research and cancer patient care. The AIFC is a multidisciplinary team of doctors, nurses, and other health care professionals from Ireland, Northern Ireland, and the U.S. whose mission is to champion the proactive management of cancer-related fatigue. The major goals of the symposium were to better understand the status and prevalence of cancer fatigue on the island of Ireland and to learn how to better assess and manage fatigue in cancer patients. An international faculty presented on topics ranging from defining the condition to developing an international protocol implementing a fatigue algorithm. PMID- 12626778 TI - Divergent effects of leptin on luteinizing hormone and insulin secretion are dose dependent. AB - We have shown recently that fasting permits leptin to modulate both luteinizing hormone (LH) and insulin secretion in cows. In rodents, leptin causes divergent effects on LH and insulin release that are dose dependent. To test the hypothesis that leptin effects on LH and insulin secretion in fasted cows are dose related, we examined the effects of various doses of recombinant ovine leptin (oleptin) in mature cows. Twenty ovariectomized beef cows, each bearing an estradiol implant to maintain basal estradiol concentrations, were used. All cows were fasted for 60 hr with free access to water and were assigned randomly to one of four groups (n = 5/group): 1) saline control; 2) leptin, 0.2 microg/kg; 3) leptin, 2.0 microg/kg; and 4) leptin, 20 microg/kg body wt. Blood samples were collected at 10-min intervals for 6 hr on Days 0 and 2, with saline or oleptin injected intravenously immediately after the first intensive sample on Day 2 (54 hr). Leptin caused a dose-related increase (P < 0.001) in mean concentrations of circulating LH. Stimulation of LH release by leptin was significant at the lowest (141% of control) and middle (122% of control) doses used, but no increase was observed for the highest dose. Increased mean concentrations of LH appeared to result from an augmentation of basal secretion, as pulse characteristics were not affected. After 54 hr of fasting, plasma insulin concentrations were lowered (P < 0.01) in all treatment groups compared to Day 0. After leptin injections, plasma insulin concentrations increased (P < 0.01) and reached highest concentrations during the first hour of sampling. However, this increase was sustained for several hours only in the intermediate (2.0 microg/kg) dose group. Collectively, our results show that leptin has potent positive effects on both LH and insulin secretion in fasted cows, but the anterior pituitary and endocrine pancreas appear to become downregulated in the presence of excess ligand. PMID- 12626780 TI - A survivor's journey: one woman's experience with cancer-related fatigue. AB - Maureen Gilbert was diagnosed at age 45 with breast cancer in Dublin, Ireland in 2000. She had a mastectomy with immediate breast reconstruction, chemotherapy, and a second prophylactic mastectomy with reconstruction. Soon after diagnosis, she began to experience fatigue that intensified to incapacitating levels with chemotherapy and stayed with her for 18 months. This experience is described in compelling terms. Fatigue was not mentioned by any of her caregivers when they spoke of symptoms and side effects. She is convinced that if the condition of fatigue had been described and validated as a symptom from the beginning, it would have helped her considerably to manage and live through it. There were some very positive outcomes from the whole experience of having cancer, but the journey would have been easier, had someone told her about fatigue. PMID- 12626782 TI - Treating fatigue in cancer patients. AB - Possible causes of cancer-related fatigue include depression, pain, sleep problems, anemia, deconditioning, metabolic abnormalities, infection, dietary problems, hypoxia, and side effects of medication. Although treatments are available for each of these conditions, there are no generally accepted treatments available for the whole fatigue syndrome. There are also very few studies on the treatment of cancer-related fatigue-only 10 randomized controlled trials. Health care providers have begun to understand that, just as the treatment of pain requires attention to imbalances in mind, body, and spirit, the treatment of fatigue will require such an approach. PMID- 12626781 TI - Fatigue and quality of life: lessons from the real world. AB - The impact of fatigue on the quality of life of oncology patients is substantial and under-recognized. Fatigue in these patients may begin with a simple decrease in physical activity, but can progress to include a wide range of negative effects that often culminate in patients feeling out of control, lonely, and isolated. In general, surviving cancer patients experience some limitations after the end of treatment but ultimately attain a reasonably good level of functioning. An examination of subpopulations and further analyses of data suggest, however, four different recovery patterns. Patients may: A) improve in their functioning, reach a plateau at approximately year 2 or 3, and then remain at relatively high levels of functioning; B) improve initially, but deteriorate again after year 2 or 3, never reaching the normal stage; C) improve, returning to normal; or D) have a very mixed pattern of high levels of fatigue that is, to date, very difficult to interpret. Disturbingly, 60% of the survivors in our population of patients with Hodgkin's disease, who were treated in recent trials of the German Hodgkin Study Group and the European Organization for Research and Treatment of Cancer Lymphoma Group, had medium to high levels of fatigue after 5 cancer-free years. Investigations are essential to determine the current status of long-term survivors in more detail and to link that status to conditions observed during the treatment of acutely ill patients. PMID- 12626783 TI - The cellular biology of erythropoietin receptors. AB - Long thought to act only as a hormone that was the primary regulator of red blood cell production, erythropoietin is now known to have a whole spectrum of activity. Organs such as the brain, ovary, oviduct, uterus, and testis have erythropoietin receptors. Because erythropoietin receptors exist in the paracrine and autocrine systems, as well as the hormonal system, the beneficial effects of administering human erythropoietin are likely to extend beyond its effect in raising hematocrit. It may even be possible in the future to use the receptors to target a drug to a tumor without damaging the surrounding tissue. PMID- 12626784 TI - Anemia in cancer: some pathophysiological aspects. AB - More than 30% of cancer patients experience anemia and its side effect, fatigue. Its causes can be numerous, but anemia is usually secondary to an imbalance of cytokines. Among these, tumor necrosis factor-alpha seems to be the major culprit, creating anemia by blunting the physiological effect of erythropoietin. Pharmacologically increasing the erythropoietin level corrects the anemia in about half the treated patients. Several studies have shown that quality of life is substantially improved through such therapy. PMID- 12626785 TI - The status of cancer fatigue on the Island of Ireland: AIFC professional and interim patient surveys. AB - The objectives of the All Ireland Fatigue Surveys are: A) to learn more about the nature of cancer patients' fatigue; B) to explore the impact that fatigue has on the lives of these patients; C) to determine physicians' and nurses' perceptions of fatigue in patients; and D) to find the current level of management of cancer fatigue in Ireland. In two surveys, one involving 109 physicians and another involving 160 nurses, the vast majority of respondents reported that they believe nausea to be the side effect of most concern to their cancer patients. In contrast, 41% of 143 patient respondents to a third survey reported that fatigue was the side effect that had the greatest impact on them. Patients indicated that fatigue affected them physically, mentally, emotionally, and economically. Yet, approximately half the patients who reported their fatigue to their physician or nurse received no treatment to relieve their fatigue. The number one recommendation made to patients who did receive treatment to help reduce their fatigue was rest/relaxation, even though research suggests that exercise can be of greater benefit. Thus, it appears that cancer patients' fatigue has been largely under-recognized and poorly managed or ignored. With the formation of interdisciplinary groups of health care professionals focused solely on cancer fatigue, efforts are under way on the island of Ireland to address this too long misunderstood and neglected aspect of cancer patients' lives. PMID- 12626786 TI - Cancer fatigue: the way forward. AB - Research in cancer-related fatigue lags far behind research in cancer-related pain and is astonishingly underdeveloped given the magnitude of the problem among cancer patients. This was recently recognized at the State-of-the-Science Conference on Symptom Management in Cancer: Pain, Depression, and Fatigue, held at the U.S. National Institutes of Health in July 2002. The results of patient surveys on fatigue being conducted in Ireland by the All Ireland Fatigue Coalition (AIFC) closely mirror results from similar surveys conducted in the U.S., in some regards, with 53% of patients experiencing significant fatigue at least daily, and 80% at least monthly on both sides of the Atlantic. In other respects, such as duration of bouts of fatigue, how much and with whom patients discuss their fatigue symptoms, and economic impacts of fatigue, there are significant differences between survey findings in the U.S. and in Ireland. But in both places, the condition is underreported and undertreated and takes an enormous toll. Work by the AIFC to document the problem in Ireland, expand public awareness, and conduct research on which to base guidelines for diagnosis and treatment is an excellent beginning to tackling this long understudied condition that afflicts the overwhelming majority of cancer patients. PMID- 12626788 TI - Understanding the process of care for musculoskeletal conditions--why a biomedical approach is inadequate. PMID- 12626789 TI - Breaking the rules: the unconventional recognition of HLA-B27 by CD4+ T lymphocytes as an insight into the pathogenesis of the spondyloarthropathies. AB - Despite extensive research, it remains unclear why a small proportion of HLA- B27(+) individuals develop spondyloarthropathies (SpA). Because the function of HLA-B27, as a major histocompatibility complex (MHC) class I molecule, is peptide presentation to CD8(+) T cells, research has concentrated on the role of HLA-B27 as a restriction element for CD8(+) cytotoxic T lymphocytes in pathogenesis. However, findings in the B27-transgenic animal models, together with the identification of unusual processing and presentation features of HLA-B27, have raised alternative hypotheses for the pathogenic role of HLA-B27. One such hypothesis is that HLA-B27 can be recognized by CD4(+) T lymphocytes. Here we report the identification of such unusual cells, which break the conventional rules of MHC restriction, and propose a model for the role of such CD4(+) T cells in SpA. PMID- 12626790 TI - Giant cell arteritis: epidemiological clues to its pathogenesis and an update on its treatment. AB - Giant cell arteritis (GCA) is a chronic systemic vasculitis with a marked female predominance and restriction to old age. The disease process distinctly targets large and medium sized arteries, preferentially the aorta and its extracranial branches. Morphological observations indicate that the age and sex distribution of GCA is related to the occurrence of degenerative changes in the arterial wall. GCA is not a truly infectious vasculitis. However, an infection might be a triggering factor. Different centres report an increase in GCA incidence, but annual fluctuations have not been shown to be statistically significant. However, significant seasonal variations have been observed by several groups. The mortality is not increased in adequately treated patients. Although, alternative steroid-sparing agents have been proposed, corticosteroids are still the first treatment choice. PMID- 12626791 TI - Clinical and psychological outcome from a randomized controlled trial of patient initiated direct-access hospital follow-up for rheumatoid arthritis extended to 4 years. AB - BACKGROUND: Patients with rheumatoid arthritis (RA) are traditionally seen regularly as out-patients, irrespective of whether it is appropriate or timely to see them. A randomized controlled trial has shown that over 2 yr, seeing patients only when they or their general practitioner (GP) request a review saves time and resources and is more convenient. This study aimed to assess clinical and psychological outcomes when the trial was extended to 4 yr. METHOD: A total of 209 patients were randomized into either 'routine review' (control) or 'no routine follow-up' but access to rapid review on request (direct access). Clinical and psychological status and patient satisfaction and confidence were reviewed after 24 and 48 months. RESULTS: Mean age at entry was 56 yr and mean disease duration 11 yr, and 134 patients remained in the study after 48 months. There were no differences between the groups, nor between those who completed the study and those who did not. There were no major differences in clinical or psychological status between the groups at 24 or 48 months. However, self efficacy for function was stronger at 48 months for direct access patients (mean 64.0 vs 52.0, P=0.005), as was self-efficacy for other symptoms (mean 67.8 vs 59.3, P=0.009). Satisfaction at 48 months was increased in direct access compared with control (mean 8.7 vs 7.6, P=0.01) as was confidence in the system (8.9 vs 7.6, P<0.01). CONCLUSION: It is effective for patients with rheumatoid arthritis to have no regular follow-up, provided they have access to rapid review when they or their GP request it. Patients using a self-referral system of care had higher self-efficacy and greater satisfaction and confidence than those using the traditional system. PMID- 12626792 TI - Psychometric properties of a Dutch short form of the Arthritis Impact Measurement Scales 2 (Dutch-AIMS2-SF). AB - OBJECTIVE: To evaluate the reliability and validity of a Dutch version of the Arthritis Impact Measurement Scales 2 short form (AIMS2-SF) and examine the agreement between the AIMS2 and AIMS2-SF in rheumatoid arthritis (RA) patients. METHODS: Data were collected from 587 RA patients from three studies. Patients completed the Dutch-AIMS2, Modified Health Assessment Questionnaire (M-HAQ), and Visual Analogue Scale for pain (VAS-pain), and clinical data were collected to calculate the Disease Activity Score 28 (DAS28). Short-form component scores were calculated from the AIMS2 long-form data. In addition, a Modified Symptom component score was calculated by replacing item 42 with item 38 as was suggested by Haavardsholm et al. [7] for the Norwegian version. RESULTS: The internal consistency of the Physical, Symptom and Affect components was good (Cronbach's alpha= 0.75-0.87), moderate for the Role component (alpha=0.62) but rather low for the Social Interaction (0.51) component. Replacing item 33 with item 31 of the long-form AIMS2 increased internal consistency for the Social Interaction component to 0.63. Test-retest reliability of the AIMS2-SF components was high (intraclass correlation coefficients >0.70). Mean scores of the AIMS2-SF were generally close to those from the AIMS2, but the limits of agreement were rather wide. Both the Modified Symptom and Modified Social Interaction components showed better agreement than the original short-form components. Plots of differences between AIMS2 and AIMS2-SF against the mean of the two scores for the five components showed that the differences varied over the range of the measurements. Factor analysis confirmed the three-factor structure, with a physical, psychological and social dimension that has been found for the Dutch-AIMS2 long form. Correlations of the AIMS2-SF components with M-HAQ total score, functional class, VAS-pain and DAS28 were very similar to the correlations for the original AIMS2. CONCLUSION: The Dutch-AIMS2-SF, with Modified Symptom and Social Interaction components has good psychometric properties, similar to those of the Dutch-AIMS2 long form. PMID- 12626793 TI - Long-term efficacy of a cognitive behavioural treatment from a randomized controlled trial for patients recently diagnosed with rheumatoid arthritis. AB - OBJECTIVES: This study examined the long-term efficacy of a cognitive behavioural intervention for patients with recent-onset, seropositive rheumatoid arthritis (RA). METHODS: Fifty-three consecutive patients with less than a 2-yr history of classic or definite RA were recruited into the trial. All participants received routine medical management during the study, and half were randomly allocated to receive an 8-week adjunctive psychological intervention. All assessments were conducted blind to the allocation. This paper reports intention-to-treat analyses of the 18-month follow-up. RESULTS: Consistent with short-term results, significant differences were found between the groups in depressive symptoms. The intervention group maintained improvements in joint function, although those in routine care made similar improvements over the ensuing 18 months. At follow-up, group differences emerged for disability and anxiety. CONCLUSIONS: These results indicate that cognitive behavioural intervention offered as an adjunct to standard clinical management early in the course of RA is efficacious in producing improvements in both psychological and physical indices. Furthermore, improvements appear to increase 18 months after a brief, time-limited psychological treatment. PMID- 12626794 TI - Interferon-gamma induces expression of interleukin-18 binding protein in fibroblast-like synoviocytes. AB - OBJECTIVE: To investigate expression of the endogenous antagonist of interleukin 18 (IL-18) bioactivity, IL-18 binding protein isoform a (IL-18BPa), in fibroblast like synoviocytes (FLS). METHODS: Long-term cultured FLS from rheumatoid arthritis (RA), osteoarthritis (OA) and spondylarthropathy patients were analysed for spontaneous and cytokine-induced IL-18BPa expression. Messenger RNA and release of IL-18BPa were assessed by semi-quantitative and quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR) as well as immunoblot analysis, respectively. RESULTS: All investigated FLS cultures expressed low amounts of IL-18BPa transcripts. However, there was no detectable release of IL 18BPa from unstimulated synoviocytes. Of the investigated cytokines, only interferon (IFN)-gamma markedly up-regulated IL-18BPa mRNA levels. Induction was accompanied by release of IL-18BPa immunoreactivity from FLS. Conditioned media from IFN-gamma-stimulated FLS cultures reduced IL-12/IL-18-dependent IFN- production by peripheral blood mononuclear cells. CONCLUSION: The present data imply that IFN--activated synoviocytes mediate a negative feedback loop via IL 18BPa, which may limit IL-18 biological activity in arthritis. PMID- 12626795 TI - Analysis of polymorphisms affecting immune complex handling in systemic lupus erythematosus. AB - OBJECTIVES: Systemic lupus erythematosus (SLE) is a polygenic disorder of dysregulated inflammation. Numerous specific candidate genes have been identified and most relate to the handling of immune complexes or antigen presentation. This is consistent with the classic finding of immune complex deposition in affected end organs. We wished to examine combinatorial effects of polymorphic variants of genes involved in immune complex clearance in susceptibility to lupus. METHODS: This study examined the occurrence of polymorphisms in genes which encode proteins known to be involved in immune complex handling and clearance. Each polymorphic variant of a complement protein (C2, mannose binding protein and C4), complement receptor (CR1) or Fc receptor (FcgammaRIIA and FcgammaRIIIA) gene is known to affect function adversely. One hundred and sixty SLE patients and 212 control subjects were genotyped using polymerase chain reaction methods. RESULTS: We found an increasing association of SLE with increasing numbers of gene defects. Combinations of severe defects in FcgammaRIIA and FcgammaRIIIA were particularly deleterious for both African American and Caucasian patients, even though only one defective variant was individually statistically significantly associated with SLE. CONCLUSIONS: The results of the study suggest that genes may interact in ways that either synergize or modify the effect of a single genetic effect and imply that association studies must be interpreted within the genetic background of the populations. PMID- 12626796 TI - Studies of human polyclonal and monoclonal antibodies binding to lupus autoantigens and cross-reactive antigens. AB - BACKGROUND: Systemic lupus erythematosus (SLE) is an autoimmune rheumatic disease serologically characterized by production of a variety of autoantibodies. Antibodies to double-stranded (ds) DNA are considered to be a diagnostic marker in SLE and their presence often correlates with active disease. The murine R4A anti-dsDNA antibody was found to cross-react with a peptide, D/EWD/EYS/G (R4A peptide), identified by analysing decapeptides selected from a peptide library. The R4A peptide inhibited binding of antibody to dsDNA and antibody deposition in kidneys in vivo. In other previous work, mice immunized with the peptide in a decapeptide form bound to a polylysine backbone, multiple antigenic peptide, were found to develop both anti-DNA and anticardiolipin antibodies. METHODS: To determine if human anti-DNA antibodies bind R4A peptide, we investigated the binding of monoclonal and polyclonal anti-dsDNA and anticardiolipin antibodies to the R4A peptide from patients with SLE. RESULTS: DNA binding by four immunoglobulin (Ig) G and two IgM human monoclonal anti-DNA antibodies was inhibited by the R4A peptide. While monomeric peptide was unable to inhibit affinity-purified polyclonal anti-DNA antibodies, serum anti-DNA reactivity was inhibited by an octameric form of the peptide in 10 SLE patients. CONCLUSIONS: Human anti-DNA reactivity includes the same fine specificity as that present in murine anti-DNA reactivity. Peptide binding might be a useful surrogate marker for SLE. PMID- 12626797 TI - Lower level of synovial fluid interferon-gamma in HLA-B27-positive than in HLA B27-negative patients with Chlamydia trachomatis reactive arthritis. AB - OBJECTIVES: To compare the synovial fluid (SF) concentrations of various cytokines in rheumatoid arthritis (RA) and in reactive arthritis, and to look for a correlation between cytokine levels and the presence of HLA-B27 antigen in reactive arthritis patients. METHODS: Concentrations of interleukin (IL) 10, IL 12, IL-18, interferon gamma (IFN-gamma) and tumour necrosis factor alpha (TNF alpha) were determined by commercially available enzyme-linked immunosorbent assays (ELISA) in the SF from 48 patients with reactive arthritis, 33 with RA and 13 with osteoarthritis (non-inflammatory controls). RESULTS: The SF concentrations of IL-10 were significantly lower in patients with reactive arthritis (median 2.3 pg/ml) than in RA patients (median 14.6 pg/ml). The SF levels of IFN-gamma were not significantly different but the ratios of IFN-gamma to IL-10 were significantly higher in patients with reactive arthritis (median 9.2) than in RA patients (median 0.83). When the subset of patients with Chlamydia trachomatis reactive arthritis was considered, the SF concentration of IFN-gamma was significantly lower in HLA-B27-positive (median 2.9 pg/ml) than in HLA-B27-negative patients (median 42.4 pg/ml). After 2 yr of follow-up, two HLA B27-positive patients, who had low SF levels of IFN-gamma, had a chronic course of arthritis, whereas after 1 yr all HLA-B27-negative patients had complete resolution of arthritis. CONCLUSIONS: The lower IFN-gamma concentrations in HLA B27-positive patients with C. trachomatis reactive arthritis could be related to the tendency of these patients to have more severe or chronic arthritis. PMID- 12626798 TI - The elevated prevalence of apolipoprotein E2 in patients with gout is associated with reduced renal excretion of urates. AB - OBJECTIVE: Previous studies have demonstrated that the lower renal excretion of urates in patients with hyperuricaemia is inversely related to plasma very low density lipoprotein (VLDL) levels, and the different genotypes of the apolipoprotein E gene are related to the plasma levels of lipids. The aim of this study was to determine the prevalence of apolipoprotein E in hyperuricaemic patients and to investigate whether the renal excretion of urates is conditioned by the apoliprotein E genotype. METHOD: The plasma levels of lipoproteins, cholesterol, triglycerides and uric acid, and the renal excretion of uric acid were studied in 68 patients with gout and in another control group of 50 healthy subjects. Both groups were genotyped for apolipoprotein E by means of an amplification technique and inverse hybridization. RESULTS: The prevalence of the E2 allele was greater in the patients than in the control group. The levels of cholesterol, triglycerides and uric acid were greater in the patients, whereas the levels of high-density lipoprotein were lower. The patients with the E2 allele had higher levels of triglycerides in VLDL and intermediate-density lipoproteins and a lower renal excretion of urates. CONCLUSIONS: These results show that the reduced renal excretion of uric acid in patients with gout is mediated by high levels of VLDL and by the high prevalence of the E2 allele of apolipoprotein E. PMID- 12626799 TI - Whipple arthritis: diagnosis by molecular analysis of synovial fluid--current status of diagnosis and therapy. AB - Whipple's disease (WD) is an uncommon polysystem infectious disease. In the present report, we describe a patient who presented with a chronic illness consistent with WD and an avascular necrosis of the right hip joint. WD and its proposed causative bacillus, Tropheryma whippelii, was identified by molecular analysis (polymerase chain reaction) in bacterial DNA extracted from the synovial fluid. The diagnosis was additionally confirmed by upper gastrointestinal endoscopy and a small bowel biopsy with macrophages positive for periodic acid Schiff reagent demonstrated by light and electron microscopy. This demonstrates that WD can be diagnosed without tissue biopsy. False diagnosis of the polymorphous signs and symptoms of WD can lead to invalidism and even death, whereas correct therapy leads to a cure in most cases. Thus, the current status of diagnosis and therapy is of key importance in the treatment of WD. PMID- 12626800 TI - The value of masters educational programmes for specialist registrars in rheumatology. AB - The training of junior doctors has undergone major changes in recent years. There is now more structure, with defined assessment time points leading to a Certificate of Specialist Training. This certificate provides documentation indicating that the trainee has undergone a satisfactory period of training and that they are sufficiently competent to practise as a specialist, unsupervised. The changes have led to re-examination of the role of, and educational provision for, research training as well as clinical training. In this article we review these issues and argue that the development of masters educational programmes may help to address several concerns. PMID- 12626801 TI - Mary, Queen of Scots--a Buxton patient. PMID- 12626802 TI - Microchimerism in Sjogren's syndrome. PMID- 12626804 TI - Complement receptor expression of relevance to apoptotic cell clearance in SLE. PMID- 12626803 TI - Re: The role of ultrasonography in the diagnosis and management of idiopathic plantar fasciitis. PMID- 12626805 TI - A case of acquired partial lipodystrophy associated with POEMS syndrome. PMID- 12626806 TI - Genetic studies in rheumatoid arthritis. PMID- 12626807 TI - Anti-tumour necrosis factor therapy in ankylosing spondylitis. A need for guidelines. PMID- 12626808 TI - Hypermobility in New Zealand. PMID- 12626810 TI - Pamidronate for SAPHO syndrome. PMID- 12626811 TI - Leflunomide induced vasculitis--a dose-response relationship. PMID- 12626812 TI - Increased plasma endothelin-1 levels in fibromyalgia syndrome. PMID- 12626813 TI - The dilemma of pseudovasculitis and aortic mural thrombi: can trans-oesophageal echocardiogram solve the problem early? PMID- 12626822 TI - Food hypersensitivity and allergic disease: a new threat in India. PMID- 12626823 TI - XXXX National Conference of Indian Academy of Pediatrics, January 2, 2003, Mumbai. PMID- 12626825 TI - Gene polymorphism and folate metabolism: a maternal risk factor for Down syndrome. AB - The high birth frequency of Down syndrome (DS), trisomy 21 (T21), has been a subject of interest to the clinicians and researchers due to its complexity in phenotypic expression. In addition to the maternal age, identification of the mechanistic basis for T21 requires an understanding of the cellular-molecular events and other biochemical pathways that could promote maternal meiotic nondisjunction. Recent studies have linked the increased frequency of polymorphism of methylenetetrahydrofolate reductase (MTHFR, C677T) and methionine synthase gene (MTRR, A66G) in mothers with DS child. Based on evidence that abnormal folate and methyl metabolism can lead to DNA hypomethylation and abnormal chromosomal segregation, researchers have observed that mothers with mutation in MTHFR (C677T) and MTRR (A66G) gene have elevated levels of plasma homocysteine. This was found to be associated with a 2.6 to 2.9 fold increased risk of having child with DS compared to mothers without the mutation. Subsequent studies evaluating Italian, Irish, French, and Indian-Gujarati women could not demonstrate an association of MTHFR gene polymorphism in mothers with DS child. However, the Irish study did find an increased risk of DS associated with the MTRR polymorphism and an interactive effect of MTRR and MTHFR polymorphisms with increased risk. Interestingly, an increase in plasma homocysteine was found to be a risk factor for DS in several of the studies. Despite the differences, the published studies suggest a common theme of abnormal folate metabolism associated with increased risk of having a child with DS. These observations suggest that there seems to be a geographic variation in gene polymorphism and it could not be attributable to meiotic nondysjunction in all mothers with DS child but increased homocysteine in all different study group does suggest that there may be a gene nutritional or gene-gene or gene-nutritional-environmental factors involved in increased frequency of meiotic nondisjunction which needs transnational and multinational study design. PMID- 12626824 TI - A comparative study of maximal aerobic power of school boys of east and north east regions of India. AB - OBJECTIVE: The evaluation of physiological responses during graded ergometry in children has been proven to be useful to determine the growth and development of cardio-respiratory and musculo-energetic systems and is also essential in sports and games. In India few attempts have been made to determine the peak oxygen consumption (VO2) of children from different regions. This study aims to investigate peak VO2 and its relation to other anthropometric parameters of school boys (8 to 14 years age) from east region (ER) and north east region (NER) of India. Also, this study attempts to find out the effect of regional variation, including their growth and development in comparison with the boys of other countries. DESIGN: Boys were selected from east and north-east states of India and then subdivided according to their age. SUBJECTS: The present study was carried out on 394 boys of 8 to 14 years of age, from three different states of ER and five states of the NER on the Indian subcontinent. METHODS: The habitual physical activity, socio-cultural characteristics were assessed by standard questionnaires. The sexual maturity status (age at puberty stage 2) was calculated by standard indices. Height, body weight and skinfold thickness were assessed by standard procedures. Peak oxygen uptake (peak VO2) of boys was measured by computerized motor driven treadmill by standard procedure. RESULTS: The results showed that the body size and peak VO2 of the boys from both regions increased significantly from 8 to 14 years of age. Peak VO2 of the subjects was less than untrained Japanese, European and American boys. The weight related and lean body weight (LBW) related peak VO2 was also changed from 8 years to 14 years boys in both the regions. It was also observed that peak VO2 was significantly and negatively correlated with the sum of skin fold thickness. Body size and peak VO2 were found to be significantly higher in the ER boys than their NER counterparts at 13 and 14 years of age only. Peak VO2 remained the same up to 12 years of age and then became significantly higher in the ER boys at 13 and 14 years of age as compared with their NER counterparts. CONCLUSIONS: There was a significant difference in peak VO2 of children from ER and NER. The high VO2 may be due to late sexual maturation and higher body size of the ER boys at that age. The difference in body size and sexual maturation may be due to genetic, racial, geographical, climatic and nutritional diversity in the east )ER) and north-east (NER) regions of India. PMID- 12626826 TI - Deciduous dentition and enamel defects. AB - Two hundred eighty children including wellnourished, malnourished and infants with intrauterine growth retardation (IUGR) were examined for dental eruption and enamel hypoplasia. In malnourished and IUGR children eruption of teeth was delayed. The prevalence of enamel hypoplsia in wellnourished children was 20% being significantly higher in females as compared to males in age group 1-2 years. Enamel hypoplasia was seen in 36.6% malnourished subjects. Breast-feeding was protective against enamel hypoplasia. PMID- 12626827 TI - Survival and morbidity in extremely low birth weight (ELBW) infants. AB - We report the morbidity and mortality in extremely low birth weight neonates (ELBW) from a tertiary care hospital over seven years (1994-2000). Data regarding maternal and neonatal details was obtained from old records, computer database and medical files. Of the 12,807 live births during this period, 137 (1.07%) were ELBW infants. All of them were managed without surfactant. Overall, 67 infants (48.7%) survived to discharge. The most commonly encountered morbidities were hyperbilirubinemia(65%), respiratory distress(65%), sepsis(52%), intraventricular hemorrhage(29%), pneumonia (25%) and retinopathy of prematurity(24%). Need for resuscitation, pulmonary hemorrhage, seizures, acute renal failure, sclerema and air leak syndromes were significantly associated with mortality. Sepsis accounted for 41% of all deaths while immaturity was the second most important cause, accounting for 24% deaths. The average length of stay for survivors was 49 days (SD +/- 15.9 days) PMID- 12626828 TI - Comparison of tympanic and rectal temperatures in febrile children. AB - The present study was designed to assess the accuracy of tympanic membrane temperature (TMT) in predicting "core" body temperature and to compare rectal temperature (RT) and TMT in febrile pediatric patients with and without meningitis. Sixty children diagnosed as having meningitis by cerebro-spinal fluid (CDF) analysis formed the cases and 60 non-meningitic febrile patients, chosen as continuous enrollment, formed the controls. Rectal and ear temperatures were assessed in both groups. Ear temperature was significantly higher in cases as compared to controls. The difference between reading of ear temperature and rectal temperature was also significantly higher in cases as compared to controls. Significant correlations were seen between ear temperature and various parameters of CSF profile. PMID- 12626829 TI - Evaluation of invasive and non-invasive diagnostic modalities for Helicobacter pylori infection in children. AB - Invasive and non invasive tests for Helicobacter pylori performed on 31 children were evaluated as diagnostic modalities. Investigations included upper gastrointestinal endoscopy and endoscopic grasp biopsy (EGB) from antrum and corpus (for rapid urease test, impression smear, histology and culture), antral brushings, serum ELISA for IgG antibodies, rapid blood test, and IgG antibodies in unstimulated saliva. Our results suggested that amongst the invasive methods brush cytology was more sensitive than histology and impression smear. Best interpretation of urease test was possible at 4 hours incubation. Culture of EGB sample constitutes the most specific way to establish the diagnosis of infection but is not easy. Hence, non-invasive modalities like serum ELISA, rapid blood test and salivary ELISA can be used in children for the detection of H pylori infection. PMID- 12626830 TI - Assessment of iodine deficiency disorders in district Bharatpur, Rajasthan. AB - Iodine deficiency disorders (IDD) is a public health problem in India. A ban on the sale of uniodised salt for household consumption has been introduced in Rajasthan State since 1992. The present study was conducted in the district of Bharatpur, Rajasthan with the objective to assess the prevalence of iodine disorders in school children as no data is available on this aspect. A total of 3072 children in the age group of 6-12 years were included in the study and were clinically examined. On the spot urine samples were collected randomly from 450 children. A total of 1064 salt samples were collected randomly from the families of the children. The total goiter prevalence was found to be 7.2% in the subjects studied. It was found that the percentage of children with urinary iodine excretion <20.0; 20.0-49.9, 50.0-99.9 and 100 mcg/L and above was 1.1, 1.1, 7.8 and 90.0% respectively. The assessment of iodine content of salt revealed that 56% of the families were consuming iodised salt. The findings of the present study indicated that the population is in a transition phase from iodine deficient (as revealed by the TGR) to iodine sufficient (as revealed by the medium UIE of 200.0 mcg/L) nutriture. PMID- 12626831 TI - Neurological complications of rabies vaccines. AB - The rabies vaccines containing neural elements are used in some countries including India. We report three cases that presented with various neurological complications following the use of these vaccines. The presenting manifestations included those of encephalitis, radiculitis and acute inflammatory demyelinating polyradiculoneuropathy. These neurological complications are highlighted so that scientific evidence compels the community to discontinue the use of the neural tissue rabies vaccines. Newer generation cell culture rabies vaccines should be preferred over the neural tissue rabies vaccines for post-exposure prophylaxis. PMID- 12626832 TI - Down syndrome with transient myeloid leukemia and urological abnormality. AB - Among the various anomalies associated with Down syndrome, leukemia is quite common. The variant transient myeloid leukemia is seen almost exclusively in the Down syndrome patients. On the other hand, urological anomalies are infrequently found both in the Down syndrome and leukemia patients. We report a case who had the rare combination of a urological anomaly along with Down syndrome and transient myeloid leukemia. PMID- 12626833 TI - Radiofrequency ablation of incessant atrial tachycardia in an infant. AB - Ectopic automatic atrial tachycardia, an uncommon type of supraventricular tachycardia in children and adults, has been reported to be resistant to medical therapy unlike reentrant supraventricular tachycardia. We report a case of incessant atrial tachycardia in an infant, which was successfully treated with radiofrequency catheter ablation. PMID- 12626834 TI - Unusual manifestations of VACTERL association. AB - We report a case of VACTERL association along with unusual manifestations of pseudo-exostrophy of bladder, hemifacial microsomia and an urachal cyst communicating with the bladder. PMID- 12626835 TI - Low level of mosaicism in atypical Prader Willi syndrome: detection using fluorescent in situ hybridization. AB - Prader Willi syndrome (PWS) most commonly is due to paternal micro-deletion of 15q11-q13. Although PWS is not a rare condition, mosaic micro-deletion cases are reported rarely. FISH using PWS micro-deletion probe is the most useful method to detect deletion including mosaicism. In this report we describe a female child with clinical features of atypical PWS and FISH analysis showing mosaicism for deletion in the PWS critical region. This is first mosaic deletion case of PWS from Indian subcontinent. PMID- 12626836 TI - Ichthyosis hystrix. PMID- 12626837 TI - Reversible cutaneous hyperpigmentation in vitamin B12 deficiency. PMID- 12626838 TI - Hypocalcemia--follow up in a tertiary hospital. PMID- 12626840 TI - Intrathecal immunoglobulin in tetanus. PMID- 12626842 TI - Nimesulide vs. paracetamol: this trial needs to stand trial. PMID- 12626845 TI - Towards the genetic analysis of mulfactorial diseases: the estimation of allele frequency and epistasis. AB - OBJECTIVES: The general aim of this paper is to reactivate the original intention behind the Elston-Stewart algorithm: i.e. physiological characterisation of the effects of individual loci underlying quantitative variation. The specific aim is the estimation of allele frequency and epistasis in multifactorial genetic diseases. METHODS: In a general genetic model, the probability of disease is a sigmoid function of the number of disease alleles summed over all loci. This model has just 4 parameters: the number of loci; the population frequency of disease alleles; a threshold expressed as a proportion of disease alleles; and the slope of the sigmoid curve. Assuming 10 loci, the remaining parameters can be estimated from empirical data: population frequency of the disease, monozygotic twin concordance rates, and disease frequency in sibs of affected probands. RESULTS: For 10 typical multifactorial diseases, the estimates of allele frequency are generally high, of the order of 20%, with strong epistatic interactions between loci. It follows that the frequencies of subphenotypes specific for a single disease locus will also be high, and only about two-fold greater in affected individuals than in normal controls. CONCLUSIONS: Because of allelic heterogeneity, purely genomic approaches are unlikely to succeed in unravelling the genetics of multifactorial diseases; this will rather require articulation with physiology and the identification of biologically meaningful subphenotypes. PMID- 12626844 TI - Detection rates for genotyping errors in SNPs using the trio design. AB - One well-known approach for the analysis of transmission-disequilibrium is the investigation of single nucleotide polymorphisms (SNPs) in trios consisting of an affected child and its parents. Results may be biased by erroneously given genotypes. Various reasons, among them sample swap or wrong pedigree structure, represent a possible source for biased results. As these can be partly ruled out by good study conditions together with checks for correct pedigree structure by a series of independent markers, the remaining main cause for errors is genotyping errors. Some of the errors can be detected by Mendelian checks whilst others are compatible with the pedigree structure. The extent of genotyping errors can be estimated by investigating the rate of detected genotyping errors by Mendelian checks. In many studies only one SNP of a specific genomic region is investigated by TDT which leaves Mendelian checks as the only tool to control genotyping errors. From the rate of detected errors the true error rate can be estimated. Gordon et al. [Hum Hered 1999;49:65-70] considered the case of genotyping errors that occur randomly and independently with some fixed probability for the wrong ascertainment of an allele. In practice, instead of single alleles, SNP genotypes are determined. Therefore, we study the proportion of detected errors (detection rate) based on genotypes. In contrast to Gordon et al., who reported detection rates between 25 and 30%, we obtain higher detection rates ranging from 39 up to 61% considering likely error structures in the data. We conclude that detection rates are probably substantially higher than those reported by Gordon et al. PMID- 12626846 TI - Association studies of QTL for multi-allele markers by mixed models. AB - In this paper, we extend association study methods of both Fan et al. [Hum Hered 2002;53:130-145], in which a quantitative trait locus (QTL) and a multi-allele marker are considered for trio families, and Fan and Xiong [Biostatistics 2003, in press], in which a QTL and a bi-allelic marker are considered for nuclear families. The objective is to build mixed models for association study between a QTL and a multi-allelic marker for nuclear families with any number of offspring. Two types of nuclear family data are considered: the first is genetic data of offspring from at least one heterozygous parents, and the second is genetic data of offspring of nuclear family. (1) For the data of offspring from at least one heterozygous parents, we assume that at least one parent is heterozygous at the marker locus, and we may infer clearly the transmission of parental marker alleles to the offspring. We show that it can be used in association study in the presence of linkage. The theoretical basis is the difference between the conditional mean of trait value given an allele is transmitted and the conditional mean of trait value given the allele is not transmitted from a heterozygous parent. To build valid models, we calculate the variance covariance structure of trait values of offspring. Besides, the reduction of the number of parameters is discussed under an assumption of tight linkage between the trait locus and the marker. (2) For the data of offspring of nuclear family, we show that it can be used in general association study. In this case, the theoretical basis is the difference between the conditional mean of trait values given an allele is transmitted from a parent and the population mean. Then, we calculate variance-covariance structure of trait values of offspring. (3) Based on the theoretical analysis, mixed models are built for each type of the data, and related test statistics are proposed for association study. By power calculation and comparison, we show that, in some instances, the proposed test statistics have higher power than that by collapsing alleles to be new ones. The proposed models are used to analyze chromosomes 4 and chromosome 16 data of the Oxford asthma data, Genetic Analysis Workshop 12. PMID- 12626847 TI - Exact tests for the analysis of case-control studies of genetic markers. AB - OBJECTIVES: The association of a candidate gene with disease can be evaluated by a case-control study in which the genotype distribution is compared for diseased cases and unaffected controls. Usually, the data are analyzed with Armitage's test using the asymptotic null distribution of the test statistic. Since this test does not generally guarantee a type I error rate less than or equal to the significance level alpha, tests based on exact null distributions have been investigated. METHODS: An algorithm to generate the exact null distribution for both Armitage's test statistic and a recently proposed modification of the Baumgartner-Weiss-Schindler statistic is presented. I have compared the tests in a simulation study. RESULTS: The asymptotic Armitage test is slightly anticonservative whereas the exact tests control the type I error rate. The exact Armitage test is very conservative, but the exact test based on the modification of the Baumgartner-Weiss-Schindler statistic has a type I error rate close to alpha. The exact Armitage test is the least powerful test; the difference in power between the other two tests is often small and the comparison does not show a clear winner. CONCLUSION: Simulation results indicate that an exact test based on the modification of the Baumgartner-Weiss-Schindler statistic is preferable for the analysis of case-control studies of genetic markers. PMID- 12626848 TI - A statistically valid alternative to the TDT. AB - OBJECTIVE: To present an alternative linkage test to the transmission/disequilibrium test (TDT) which is conservative under the null hypothesis and generally more powerful under alternatives. METHODS: The exact distribution of the TDT is examined under both the null hypothesis and relevant alternatives. The TDT is rewritten in an alternate form based on the contributions from each of the three relevant parental mating types. This makes it possible to show that a particular term in the estimate is an exact tie and thus to rewrite the estimate without this term and to replace the multinomial 'variance estimate' of Spielman et al. [Am J Hum Genet 1993;52:506-516] by the binomial variance. RESULTS: The resulting test is shown to be a stratified McNemar test (SMN). The significance level attained by the SMN is shown to be conservative when compared to the asymptotic chi(2) distribution, while the TDT often exceeds the nominal level alpha. Under alternatives, the proposed test is shown to be typically more powerful than the TDT. CONCLUSION: The properties of the TDT as a statistical test have never been fully investigated. The proposed test replaces the heuristically motivated TDT by a formally derived test, which is also computationally simple. PMID- 12626849 TI - Regional cerebral blood flow patterns and response to donepezil treatment in patients with Alzheimer's disease. AB - We attempted to identify the characteristic patterns in regional cerebral blood flow (rCBF) of patients with Alzheimer's disease who responded to donepezil therapy. Sixty-one patients treated with donepezil were divided into two groups (responders and nonresponders) on the basis of changes in their Mini Mental State Examination (MMSE) scores from baseline to study endpoint. We analyzed single photon emission computed tomography data (SPECT) at baseline using three dimensional stereotactic surface projections and compared differences in rCBF between the two groups. Statistical maps showed a significantly lower rCBF of the lateral and medial frontal lobes in the nonresponders than in the responders. There was a significant inverse correlation between the relative rCBF reduction in the frontal lobe and the MMSE change. These results suggest that frontal function, as assessed by SPECT, affects the patient's response to treatment with donepezil. Measuring rCBF may aid in the selection of possible treatment responders. PMID- 12626851 TI - An observational clinical study of the efficacy and tolerability of donepezil in the treatment of Alzheimer's disease. AB - An open-label, observational Post-Marketing Surveillance (PMS) study was undertaken in Germany to examine the efficacy and tolerability of donepezil in routine clinical practice. Alzheimer's disease (AD) patients were treated with donepezil (5 or 10 mg once daily) and observed for a period of approximately 3 months. Study assessments included the Mini-Mental State Examination (MMSE), the Nurses' Observation Scale for Geriatric Patients (NOSGER), and adverse events (AEs). A total of 2,092 patients (mean age 73.0 years; mean +/- SD MMSE score 17.8 +/- 5.8) were included in the efficacy assessments. MMSE and NOSGER scores showed statistically significant improvements in the total patient population and in the subpopulations with severe AD or AD with concomitant Parkinsonian symptoms (ADPS cohort). AEs were reported in a total of 12% of patients and were mostly due to peripheral cholinergic effects. In this observational PMS study, donepezil was shown to be an effective and well-tolerated therapy in the overall patient population, in patients with severe AD, and in the ADPS cohort. PMID- 12626850 TI - Cardiovascular effects of donepezil in patients with dementia. AB - Heart rate variability is used to assess cardiovascular autonomic function. The cholinesterase inhibitor donepezil potentially affects parasympathetic activity. Twenty participants with Alzheimer's disease or dementia with Lewy bodies were treated with donepezil in a pilot study. Power spectral analysis was used to analyse 5 min of beat-to-beat RR interval data in 15 cases. Heart rate variability was significantly reduced following treatment with donepezil; mainly for high frequency (median changed from 581 to 78 ms2; p = 0.001) but also for total power (median changed from 1,563 to 844 ms2; p = 0.047). Donepezil may adversely influence cardiovascular autonomic control. These results indicate the need for larger controlled trials to further investigate the cardiovascular effects of donepezil. PMID- 12626852 TI - Construct validity of the abbreviated mental test in older medical inpatients. AB - OBJECTIVES: To evaluate validity and internal structure of the Abbreviated Mental Test (AMT), and to assess the dependence of the internal structure upon the characteristics of the patients examined. DESIGN: Cross-sectional examination using data from the Italian Group of Pharmacoepidemiology in the Elderly (GIFA) database. SETTING: Twenty-four acute care wards of Geriatrics or General Medicine. PARTICIPANTS: Two thousand eight hundred and eight patients consecutively admitted over a 4-month period. MEASUREMENTS: Demographic characteristics, functional status, medical conditions and performance on AMT were collected at discharge. Sensitivity, specificity and predictive values of the AMT <7 versus a diagnosis of dementia made according to DSM-III-R criteria were computed. The internal structure of AMT was assessed by principal component analysis. The analysis was performed on the whole population and stratified for age (<65, 65-80 and >80 years), gender, education (<6 or >5 years) and presence of congestive heart failure (CHF). RESULTS: AMT achieved high sensitivity (81%), specificity (84%) and negative predictive value (99%), but a low positive predictive value of 25%. The principal component analysis isolated two components: the former component represents the orientation to time and space and explains 45% of AMT variance; the latter is linked to memory and attention and explains 13% of variance. Comparable results were obtained after stratification by age, gender or education. In patients with CHF, only 48.3% of the cumulative variance was explained; the factor accounting for most (34.6%) of the variance explained was mainly related to the three items assessing memory. CONCLUSION: AMT >6 rules out dementia very reliably, whereas AMT <7 requires a second level cognitive assessment to confirm dementia. AMT is bidimensional and maintains the same internal structure across classes defined by selected social and demographic characteristics, but not in CHF patients. It is likely that its internal structure depends on the type of patients. The use of a sum-score could conceal some part of the information provided by the AMT. PMID- 12626853 TI - Correlation between intrathecal sulfatide and TNF-alpha levels in patients with vascular dementia. AB - OBJECTIVES: Subcortical vascular dementia (SVD) is associated with white matter lesions and demyelination. The aim of the present study was to examine the cerebrospinal fluid (CSF) levels of TNF-alpha, a proinflammatory cytokine mediating myelin damage, in SVD patients. The intrathecal TNF-alpha levels were related to the clinical symptoms of dementia, as well as to intrathecal levels of sulfatide, a marker of white matter degradation, and of neurofilament, a marker of neuronal degeneration. METHODS: CSF levels of TNF-a, sulfatide and neurofilament were all analyzed by immunoenzymatic procedures in 17 patients with SVD and in 26 healthy controls. RESULTS: The intrathecal concentration of TNF alpha was significantly increased in SVD patients compared to healthy controls (p = 0.0001). The intrathecal levels of TNF-alpha were significantly correlated (r = 0.6, p = 0.02) to the levels of sulfatide, but not to the levels of neurofilament, (r = 0.08, NS). CONCLUSIONS: We have demonstrated intrathecal production of TNF-alpha in SVD patients. The correlation between TNF-a and sulfatide levels in the CSF suggests that this apoptosis-inducing cytokine leads to the death of oligodendrocytes, thereby contributing to white matter degeneration, a hallmark of SVD. PMID- 12626854 TI - Objective measurement of the alertness level in dementia. AB - The alertness level of 143 patients with dementia was investigated. The patients were examined clinically and electroencephalographically, and the results were compared with 21 healthy volunteers. Regional brain syndromes were determined clinically. For the assessment of the EEG, a method was employed that distinguishes alertness-related slow wave activity from abnormal slow activity. The shifts of the EEG power were monitored during the 5-min examination. The short-term variations were considered to be caused by changes in alertness, whereas the basal level was supposed to reflect the EEG abnormality. It was found that a decrease in alertness occurred more often in the demented patients than in the controls. The intensity of this decrease was proportional to the degree of the dementia. The alertness disturbance was found to be particularly associated with the parietal brain syndrome. PMID- 12626855 TI - Subjective memory impairment, cognitive function and depression--a community study in older Koreans. AB - Correlates of subjective memory impairment (SMI) were investigated using data from a community study of 1,204 individuals aged 65 or over in an urban and rural area of South Korea. SMI and depression were ascertained from the Geriatric Mental State Schedule and cognitive function from the Korean version of the Mini Mental State Examination (MMSE-K). 686 participants had also completed the MMSE-K two years earlier. SMI was present in 22% of the sample and was associated with depression and lower MMSE-K scores. Depression and SMI were most strongly associated in the presence of cognitive impairment. SMI was weakly associated with previous cognitive decline and was not associated with APOE e4. SMI and cognitive impairment were most strongly associated in urban residents, particularly rural-to-urban migrants. PMID- 12626856 TI - Prevalence of dementia in China. AB - To obtain prevalence estimates of dementia in China, an analysis of 17 studies published in Chinese from 1990-1999 was carried out. The prevalence rates for the population aged 60 years and older were 1.26% for Alzheimer's disease (AD) and 0.74% for vascular dementia (VD). The prevalence of AD was 2.10% in women and 0.76% in men, while the prevalence of VD was 0.71 and 0.69%, respectively. The prevalence of AD among the three educational levels in our study (illiterate, primary school and high school) were 1.79, 0.45 and 0.15%, respectively, and those of VD were 0.26, 0.58 and 0.26%, respectively. Although the prevalence of AD (2.29%) was higher in urban than in rural areas (1.67%), the difference was not statistically significant. The difference between the prevalence of VD in urban (0.67%) and in rural areas (1.13%) was not significant either. The prevalence of AD increased with age, and gender was found to be associated with Alzheimer's disease. The prevalence of VD also increased with age, but there was no association between VD and gender. PMID- 12626857 TI - Mortality in Alzheimer's disease. AB - OBJECTIVE: This study examines the relevance of variables suspected to influence mortality in Alzheimer's disease (AD). METHODS: 172 subjects with AD recruited through a family study were followed up for a mean of 4.7 +/- 2.6 years. Their survival was compared with that of matched control subjects using Kaplan-Meier and log-rank statistics. Variables determining mortality were examined with the Cox proportional hazards model. RESULTS: Survival in subjects with AD depended on age, the severity of cognitive decline and on the incidence of hospitalisation. Gender, the duration of the illness, the velocity of cognitive decline, the allele E4 of the apolipoprotein E genotype and the presence of other lifetime psychiatric diagnoses did not influence mortality. CONCLUSION: Mortality in AD depended mainly on the subjects' age and the severity of the disorder. PMID- 12626859 TI - Neuronal classes in the isocortex of a monotreme, the Australian echidna (Tachyglossus aculeatus). AB - We have used Valverde-Golgi and Golgi-Colonnier techniques to analyze cortical neuronal morphology in four regions (frontal cortex, primary motor cortex, primary somatosensory cortex, primary visual cortex) of the isocortex of the echidna (Tachyglossus aculeatus). Eight classes of neurons could be identified- pyramidal, spinous bipolar, aspinous bipolar, spinous bitufted, aspinous bitufted, spinous multipolar, aspinous multipolar and neurogliaform. All except the pyramidal neurons were morphologically similar to neuronal classes seen in eutherian and metatherian isocortex. Pyramidal neurons made up a small proportion of all cortical neurons encountered in our preparations of echidna cortex (34% in visual cortex, 35% in somatosensory cortex, 41% in frontal cortex and 49% in motor cortex) compared to both reported values in eutherian cortex and values we found in rat cortex impregnations prepared in an identical fashion to the echidna material (75% in rat motor and 78% in rat somatosensory cortex). Many pyramidal neurons in the echidna isocortex were atypical (30-42% depending on region) with inverted somata, short or branching apical dendrites and/or few basal dendrites, very different from the usual pyramidal neuron morphology in eutherian cortex. Dendritic spine density on apical and basal dendrites of echidna pyramidal neurons in somatosensory cortex and apical dendrites of motor cortex pyramidal neurons was also lower than that found in the rat. The present findings are consistent with both pyramidal neurons and the many diverse types of non pyramidal neurons having already emerged as discrete morphological entities very early in mammalian cortical evolution, at the time of divergence of the therian and prototherian lineage. PMID- 12626858 TI - The scaling of white matter to gray matter in cerebellum and neocortex. AB - It is known that the white matter of neocortex increases disproportionately with brain size. However, relatively few measurements have been made of white matter/gray matter scaling in the cerebellum. We present data on the volumes of white and gray matter in both structures, taken from 45 species of mammals. We find a scaling exponent of 1.13 for cerebellum and 1.28 for neocortex. The 95% confidence intervals for our estimates of these two exponents do not overlap. This difference likely reflects differences in the connectivity and/or micro structure of white matter in the two regions. PMID- 12626861 TI - The capsular organ of Chelyosoma productum (Ascidiacea: Corellidae): a new tunicate hydrodynamic sense organ. AB - Chelyosoma has about 200 sense organs in the atrial wall of the branchial sac that show structural features suggestive of a role in hydrodynamic sensing. They consist of a fluid-filled capsule with an acellular diaphragm spanning an opening in the top. The floor of the capsule is a sensory macula, with 5-6 primary sensory neurons whose cilia project into the capsular cavity and whose axons go to the brain. Animals were tested for vibrational sensitivity using a loudspeaker probe. They responded by muscular contractions of the siphons and/or arrests of the branchial cilia, both of which caused changes in water flow velocity through the siphons, which were monitored non-invasively using a thermistor flow meter. The animals showed peak sensitivity at 240-260 Hz. Subsequent calibration of the loudspeaker probe indicated that the animals could detect 104 decibels re 1 microPa from a source 80 cm away. The majority of axons leaving the capsular organs go to the brain via the visceral nerve. Cutting this nerve abolished or greatly reduced responses. Electrophysiological recordings from the distal nerve stump showed bursts of electrical events following vibrational stimulation. No such records could be obtained from other major nerves and cutting them did not affect responsivity. Corella inflata, a close relative of Chelyosoma, lacks capsular organs and failed to show any responses when the source was more than 3.5 cm away. We conclude that Chelyosoma's vibration-sensing ability is due to its capsular organs and is adaptive in terms of detecting the movements of objects in the vicinity. The findings are discussed in relation to the evolution of hydrodynamic mechanoreceptors in tunicates, amphioxus and craniates. PMID- 12626860 TI - Evolution of specialized pyramidal neurons in primate visual and motor cortex. AB - The neocortex of primates contains several distinct neuron subtypes. Among these, Betz cells of primary motor cortex and Meynert cells of primary visual cortex are of particular interest for their potential role in specialized sensorimotor adaptations of primates. Betz cells are involved in setting muscle tone prior to fine motor output and Meynert cells participate in the processing of visual motion. We measured the soma volumes of Betz cells, Meynert cells, and adjacent infragranular pyramidal neurons in 23 species of primate and two species of non primate mammal (Tupaia glis and Pteropus poliocephalus) using unbiased stereological techniques to examine their allometric scaling relationships and socioecological correlations. Results show that Betz somata become proportionally larger with increases in body weight, brain weight, and encephalization whereas Meynert somata remain a constant proportion larger than other visual pyramidal cells. Phylogenetic variance in the volumetric scaling of these neuronal subtypes might be related to species-specific adaptations. Enlargement of Meynert cells in terrestrial anthropoids living in open habitats, for example, might serve as an anatomical substrate for predator detection. Modification of the connectional and physiological properties of these neurons could constitute an important evolutionary mode for species-specific adaptation. PMID- 12626862 TI - [Congenital genital anomalies. Aspects of diagnostics and treatment]. AB - Congenital genital anomalies are a very complex pathology. In order to clarify its causes it is important to revert to the genetic conditions and regularities of embriological development. The genital disturbances are mostly determined by chromosomal or endocrinic disorders or by impaired biochemical processes. Clinical problems arise when the genetical sex is in discrepancy with ambiguous genitalia. True hermaphroditism, congenital adrenal hyperplasia, testicular feminization and gonadal dysgenesis are the most common syndromes. Diagnostic criteria applied are similar for all (establishment of karyotype, investigation of hormones and their derivates, genital ultrasound and endoscopy, if needed - radiological examination), but medical and surgical treatment is applied to each patient individually. PMID- 12626863 TI - [Mental disorders and their relation to brain lesion location: diagnostic problems]. AB - Knowledge of symptoms of appropriate brain areas lesion helps to differ psychiatric and neurological disorders. The objective of our work was to find out the situation in scientific research about mental disorder relation to brain lesion location and to except the location of lesions, which are most complicated in differential diagnosis. We discussed the relation of most important mental disorders to brain lesion location. The study of discrete organic cerebral lesions resulting in clearly definable psychiatric disorders may provide an understanding of the underlying pathophysiological basis of these disorders. Different nervous functions need the integrational work of various brain areas. The regions differ from each other by the importance of playing part in corresponding functions. The differential problems appear because various structural brain lesions provide symptoms, similar to mental disorder symptoms. The development of mental disorders and lesion location questions are very urgent. While analyzing the location of lesion, it is important to motivate the theories of development of schizophrenia, organic depression, emotional lability and other disorders. PMID- 12626864 TI - [Relationship between lower extremity muscle strength and dynamic balance in people post-stroke]. AB - The purpose of this study was to determine how lower extremity muscle weakness is related to post-stroke difficulties in balancing. METHODS: Dynamic balance of 30 people post-stroke and 30 neurologically sound people was assessed by the Functional Reach Test and the Timed Up and Go Test. Bilateral lower extremity muscle strength was measured in classical manual muscle testing positions using a Lafayette instrument. RESULTS: There was a weak correlation between lower extremity muscle strength and the Functional Reach Test: from r=0.05 to r=0.53 for the impaired extremity and from r=0.23 to r=0.53 for the sound extremity. Control group results were from r=0.51 to r=0.86. The correlation between lower extremity muscle strength and the Timed Up and Go Test was from r=-0.33 to r= 0.64 for the impaired extremity and from r=-0.35 to r=-0.58 for the sound extremity. Control group results in this testing situation were from r=-0.63 to r=-0.90. CONCLUSION: The results of the study indicate that the loss of lower extremity muscle strength as a result of cerebrovascular accident has a poor influence on dynamic balance problems in people post-stroke. PMID- 12626865 TI - [Comparison of two different methods of analgesia. Postoperative course after colorectal cancer surgery]. AB - BACKGROUND: The purpose of our study is to compare two methods of postoperative analgesia in colorectal cancer patients after resectional operations, and to evaluate advantages and limitations of each method on the postoperative course of these patients. METHODS: One hundred patients scheduled to undergo elective colorectal cancer surgery were randomized into two groups; after general anesthesia, one group received epidural analgesia (n=50) and the second one - intramuscular pethidine analgesia (n=50). Visual analogue scale at rest and on coughing was used to compare intensiveness of pain between the two groups during the day of surgery and first three postoperative days. Patients' mood and self satisfaction were evaluated using self-assessment manikin scale. Side effects of both analgesia techniques were registered. All complications and postoperative hospital stay were also evaluated. RESULTS: Visual analogue scale pain scores at rest and on coughing were significantly better in epidural analgesia group as compared to systemic intramuscular pethidine analgesia group (p<0.05). Additional analgesics were needed for 10 (20%) and 28 (56%) patients respectively to keep visual analogue scale pain scores below 5. Adverse effects such as profound sedation, nausea and vomiting were more frequent in systemic intramuscular pethidine group, but pruritus - very uncommon to compare with epidural analgesia group (p<0.05). There were no significant differences between the two groups in respect to complications and postoperative hospital stay. CONCLUSIONS: Epidural analgesia has demonstrated significantly better effectiveness than intramuscular pethidine analgesia after colorectal cancer surgery with fewer adverse events. Self-assessment manikin scores showed better self-satisfaction in patients of epidural analgesia group as compared to patients in systemic pethidine group. PMID- 12626866 TI - [Early diagnosis of genital Chlamydia trachomatis infection among adolescent girls]. AB - Genital Chlamydia trachomatis infection is the most frequently found sexually transmitted disease among sexually active adolescent 15-19-year-old girls. In general, infection caused by C. trachomatis tends to be less abrupt in onset, clinical course is usually characterized as asymptomatic or with mild symptoms. If is although the cause of moderate to severe complications (infertility, pelvic inflammatory diseases) at a later age. OBJECTIVE: To study the prevalence of chlamydia infection and the possibilities and of early detection among adolescent girls. METHODS: Four hundred eighty seven 15-19-year-old girls were questioned by anonymous questionnaire. All 240 sexually active girls were advised to come for the investigation to the Clinic of Pediatrics of Kaunas University of Medicine Hospital. Fifty nine girls underwent inspection of the genitalia, had a speculum and bimanual pelvic examination, test of CT/NG DNA hybridisation for C. trachomatis from vagina. Smears from urethra vagina and cervix were taken. RESULTS: Test for C. trachomatis infection was positive in 11 (18%) by CT/NG DNA hibridisation from vagina. Detection of bad smelling discharge from vagina together with nonspecific changes in vaginal and cervical mucosa, mount and methylene blue prepared smears could reliably predict the diagnosis of C. trachomatis infection. CONCLUSION: We found high prevalence of C. trachomatis infection among adolescent girls. Only clinical symptoms together with nospecifical test were reliable predictors of the final diagnosis, enabling to start the early treatment. PMID- 12626867 TI - Results of treatment of extrahepatic cholangiocarcinoma at Kaunas University of Medicine Hospital. AB - Adenocarcinoma of the extrahepatic bile ducts is considered to be a rare cause of obstructive jaundice. Prognosis is poor and only radical surgery can prolong the life of such patients. The aim of study was to evaluate the outcome of treatment of patients having extrahepatic carcinoma of the bile ducts. Patients underwent treatment at Kaunas University of Medicine Hospital. A retrospective study was performed of 57 patients suffering from cancer of extrahepatic bile ducts in 1996 2001. Diagnosis was established by ultrasonography, endoscopic retrograde cholangiopancreatography, percutaneous transhepatic cholangiography and computed tomography. Sixteen patients (25%) received symptomatic treatment. Thirty-three patients (61%), receiving palliative therapy, underwent a biliodigestive bypass (n=14), an intraoperative biliary stenting (n=10), an endoscopic biliary drainage (n=6), or percutaneous transhepatic biliary drainage (n=3). Eight patients (14%) underwent radical resections of bile ducts: five patients had resections of bile ducts with D2 lymphonodectomy, two patients had bile duct resections in combination with pancreatoduodenal resection and one patient had resection of the bile ducts in combination with right hepatectomy. Histological examinations of the resected specimens revealed a curative effect in seven out of eight cases. There were no deaths in the group of radically treated patients. In the group of patients receiving palliative therapy 8 patients died: four patients died because of cardiovascular complications, two patients died of hepatorenal syndromes, one patient died because of multiorgan failure and one died of peritonitis. Five patients died in the group receiving symptomatic treatment. Assessing remote results of treatment, median survival time of patients who underwent resection was 89 days. Survival results of patients who had curative resections were much better (ranging from 277 to 1084 days). Median survival time of patients, receiving palliative treatment, was 65 days and 29 days of group, receiving symptomatic treatment. Recent progress made in surgical techniques and in perioperative management of extrahepatic cholangiocarcinomas as well as modern diagnostic achievements has greatly contributed to the outcomes of radical surgery. The main aim of the treatment is to perform resection of the bile ducts cancer. PMID- 12626868 TI - [The impact of morphologic and physiologic peculiarities of the pancreas on pancreas-related complications following pancreatoduodenectomy]. AB - BACKGROUND: The factors influencing failure of pancreaticojejunostomy following pancreatoduodenectomy are still ill-defined. Our previous study showed that age of patient, bilirubinemia or malignant nature of peripancreatic tumor had no impact on pancreas-related morbidity following pancreatoduodenectomy. The hypothesis is that it could be influenced by the level of pancreatic fibrosis, diameter of the main pancreatic duct, and exocrine pancreatic function. Aim of the study was to analyze the impact of morphologic and physiologic peculiarities of pancreatic remnant on development of pancreas-related morbidity after Whipple procedure. MATERIAL AND METHODS: We have analyzed retrospectively clinical data of 122 patients who have undergone pancreatoduodenectomy in the Department of Surgery of Kaunas University of Medicine Hospital during 1995-2001. Fibrosis of pancreatic parenchyma, diameter of main pancreatic duct, and preoperative exocrine function were evaluated. Pancreas-related morbidity was determined as either peripancreatic sepsis or pancreatic fistula. Fibrosis of pancreatic remnant was determined by computer-aided morphometric analysis. The exocrine pancreatic function was tested the day before surgery by Pancreatic Elastase-1 Stool test. RESULTS: One hundred twenty two patients have undergone pancreatoduodenectomy during 1995-2001. Pancreas-related morbidity was encountered in 27 (22.13%) cases, pancreatic fistula in 13 (10.65%) and peripancreatic sepsis in 14 (11.47%). Univariate analysis shows that diameter of main pancreatic duct and level of postoperative amylasemia were significantly different between the groups with and without pancreatic complications (p=0.001 and p=0.002, respectively) as well as there was significant difference of pancreatic exocrine function and fibrosis between the groups of patients who developed pancreas-related complications and who did not (p=0.003 and p=0.026, respectively). When logistic regression analysis was applied on those 4 variables, only one independent risk factor - exocrine pancreatic function at the cut-off of stool Elastase 100 micro g/g was revealed (odds ratio 21.6). The sensitivity of the Stool Elastase-1 test was 0.86, specificity 0.78, positive predictive value - 0.55 and negative predictive value - 0.95. CONCLUSIONS: The level of pancreatic fibrosis, diameter of the main pancreatic duct, and exocrine pancreatic function mainly influence pancreas-related morbidity following pancreatoduodenectomy. Exocrine pancreatic function measured by Stool Elastase-1 test is helpful for the detection of the group of patients with minimal risk for pancreas-related morbidity after pancreatoduodenectomy. PMID- 12626869 TI - [Emergency thoracotomy]. AB - OBJECTIVE: To determine indications for emergency (immediate and urgent) thoracotomies in cases of penetrating and blunt chest traumas. METHODS: We performed retrospective analysis of treatment methods and results, achieved in 2927 patients treated for chest traumas in 1987-2000. RESULTS: Algorithms for decision making in surgical management of chest traumas are drawn. Indications for emergency (immediate and urgent) thoracotomies are determined. Immediate thoracotomy was performed in 17.2% and 0.2% of patients, suffering from penetrating and blunt chest trauma respectively. Urgent thoracotomy underwent 7.6% due to penetrating and 2.7% due to blunt chest trauma. Postoperative mortality rate was much higher after immediate (20.0%) and urgent (10.9%) thoracotomy performed due to blunt chest traumas. In cases of penetrating chest traumas postoperative mortality rate was 3.3% and 3.0% after immediate and urgent thoracotomies respectively. CONCLUSIONS: The effectiveness of surgical treatment of chest traumas depends on logical determination of indications for immediate or urgent thoracotomies. They should be undertaken in cases of severe damage of chest wall and/or internal organs and dangerous pathological syndromes. PMID- 12626870 TI - [The diagnostic value of myocardial perfusion in diagnosis of ischemic cardiomyopathy and hypertensive heart disease]. AB - Objective of this study was to detect regional myocardial perfusion defects performing (99m)Tc-MIBI myocardial perfusion imaging and to compare the results with echocardiography for differential diagnostics of the ischemic and hypertensive cardiomyopathy until coronary angiography will be performed. MATERIAL AND METHODS: In total 50 patients with cardiomegaly have been evaluated: 18 patients with hypertensive cardiomyopathy (I(st) group), 15 - with ischemic and hypertensive cardiomyopathy (II(nd) group) and 17 patients with ischemic cardiomyopathy (III(rd) group). All patients underwent 2D echocardiography examination and (99m)Tc-MIBI myocardial perfusion imaging before coronary angiography was done. RESULTS: Thickness of interventricular septum, myocardial mass and relative wall thickness were statistically significantly smaller in the III(rd) group of patients in comparison with the I(st) and the II(nd) group of patients. CONCLUSIONS: Logistic regression model including selected data from myocardial perfusion imaging with (99m)Tc-MIBI in combination with selected echocardiography data enables prognosis of coronary arteries stenosis with 91.2% sensitivity and 93.8% specificity. PMID- 12626871 TI - [Streptococcus bovis endocarditis - predictor of colonic carcinoma? ]. AB - The association of Streptococcus bovis endocarditis with colonic neoplasia is classically described and exceeds 50 percent. The prevalence of Streptococcus bovis in fecal cultures from patients with carcinoma of the colon is significantly increased (56%) as compared to that in controls (10%) and also in patients with inflammatory bowel disease (28%). Streptococcus bovis endocarditis is relatively benign, but it stresses the frequency of the associated colonic carcinoma, requiring colonoscopy and making the treatment of the high-risk lesion mandatory. The article covers a literature review and a case report. PMID- 12626872 TI - [Development of prodioxin ointment]. AB - There was a purpose to create a composite ointment of proteolytic and antimicrobial activity, the formulation of which would contain the proteolytic enzyme procelan and the antimicrobial preparation with wide spectrum of action dioxidin. During the development of prodioxin ointment by means of biological experiments the optimal concentration of dioxidin was evaluated; it was 1 percent. The optimal dispersity value of dioxidin particles was estimated by dialysis through a semipermeable membrane. The best release of dioxidine from prodioxin ointment occurred, when it was pulverized to particles with diameter 5 10 mm. The method of assay of dioxidin was modified for the ointment. The physicomechanical properties of the prodioxin ointment: flowability, colloidal stability, thermostability and rheological properties were examined. The investigated ointment is non-newtonian in character, e.g. an elastic and plastic system and may be intended for use in dermatological practice. Medicinal enzymic and antimicrobial effects of the prodioxin ointment were compared with the healing effect of Iruksol ointment. The medicinal effect of the prodioxin ointment exceeded that of Iruksol ointment by one day. Judging by the results of the physicochemical analysis a conclusion can be made that the prodioxin ointment is stable for two years at room temperature. PMID- 12626873 TI - [Personality correlations with depressiveness among adolescents]. AB - OBJECTIVE: The aim of the study was to assess personality correlates of depressiveness as well as their relationships with the level of depressiveness. MATERIAL AND METHODS: The data on the depressiveness of 954 (452 boys and 502 girls) of ninth grade schoolchildren from 26 secondary schools of one administrative region of Lithuania were analyzed. The methods used in the study were Minnesota Multiphasic Personality Inventory for Adolescents, and Composite International Diagnostic Interview. RESULTS: Twenty two schoolchildren (5 boys and 17 girls) with extremely high scores on Minnesota Multiphasic Personality Inventory Depression scale were found to have depression or dysthymia. It was found that depressive boys and girls (high scores on scores Minnesota Multiphasic Personality Inventory Depression scale) had higher scores on Minnesota Multiphasic Personality Inventory clinical scales (except Mania scale, which was lower) as compared to the non-depressive ones. Girls with depressiveness, which corresponded to the level of depression or clinical dysthymia by Composite International Diagnostic Interview had higher scores on Minnesota Multiphasic Personality Inventory clinical scales (except Mania scale, which was lower), as compared to the other girls (both depressive and non-depressive, as measured by Minnesota Multiphasic Personality Inventory Depression scale). Girls with depressiveness, which corresponded to the level of clinical depression or clinical dysthymia, also had worse psychological adjustment. CONCLUSION: Depressive adolescents, boys and girls, differ from non-depressive ones by worse psychological adjustment. The level of depression did not differ from depressive girls (having high scores on Minnesota Multiphasic Personality Inventory Depression scale) in their non-adaptive personality characteristics, but markedly differed from that of other girls irrespective of theirs Minnesota Multiphasic Personality Inventory Depression scale scores. PMID- 12626874 TI - [The investigation of the condition of oral cavity among the schoolchildren of 5th-6th grade]. AB - The data of dental caries prevalence and intensity among schoolchildren of the fifth-sixth grade of six schools in Vilnius were analyzed. It was found that 83% of 908 investigated schoolchildren had carious teeth, 75.9% of them had dental caries in the permanent teeth and 7.1% in the milkteeth (permanent teeth were healthy). The dental caries of permanent teeth was 6.2% more frequent among the girls in comparison with the boys and was 5.8% more frequent among the Lithuanian pupils in comparison with investigated pupils of other nationalities. The prevalence of caries increased with the schoolchildren age and was 20.6% more frequent among 13-year old pupils in comparison with the 11-year old. The intensity of caries increased with the age and was higher among the pupils with bad oral hygiene, which was found in 19.5% of investigated schoolchildren. PMID- 12626876 TI - Chronological changes of N-methyl-D-aspartate receptors and excitatory amino acid carrier 1 immunoreactivities in CA1 area and subiculum after transient forebrain ischemia. AB - We investigated changes of immunoreactivities of N-methyl-D-aspartate receptor (NR) and of excitatory amino acid carrier 1 (EAAC-1), the neuronal glutamate transporter, in the vulnerable CA1 area and the less vulnerable subiculum of the gerbil hippocampus at various times following transient forebrain ischemia. At 30 min after ischemia-reperfusion, the intensity of NR immunoreactivity increased markedly in neurons of CA1 and subiculum, particularly NR2A/B, while EAAC-1 immunoreactivity was reduced in CA1. At 3 hr after reperfusion, the density of NR1 immunoreactivity markedly decreased in CA1. In contrast EAAC-1 immunoreactivity increased in CA1 and in the subiculum. At 12 hr after reperfusion, the decrease of NR1 immunoreactivity was not detected whereas EAAC-1 immunoreactivities in the CA1 area were intensified. In the subiculum, both NR subunits immunoreactivities decreased significantly, in contrast to the maintenance of EAAC-1 immunoreactivity. At 24 hr after reperfusion, both NR2A/B and EAAC-1 immunoreactivities decreased markedly in CA1 and subiculum. We tentatively suggest that the increase of NR immunoreactivity in CA1 at early times after ischemia-reperfusion may increase the delayed neuronal death, and that the increase or maintenance of EAAC-1 immunoreactivity at early times after ischemia-reperfusion may be an important factor in survival of neurons. PMID- 12626875 TI - [Preclinical studies of concomitant chemoradiation]. AB - Animal models are widely used in preclinical studies in order to explain the mechanisms of action of chemotherapy, radiotherapy and concomitant chemoradiation, to analyze pathophysiology of tumors and to evaluate the treatment of choice for malignant tumors. The choice of murine tumor or human tumor xenograft system is still debated. Xenografted human tumors have two main advantages: their human origin and wanted pathological type, which is necessary for future clinical studies. There are a lot of disadvantages of xenografted tumors: the stroma and vascular network of transplanted tumors have murine origin, the graft is mostly ectopic, the volume of transplanted tumors at the time of chemoradiotherapy is much smaller than that of the tumor in man; due to residual immunity it is difficult to determine response to cytotoxic treatment. It is still impossible to extrapolate the results obtained in a tumor model in animal to man. These investigations are usefull for interpretation of clinical results and for proposing the less empirical method of chemoradiation for phase II clinical trials. PMID- 12626877 TI - Differential distribution of aldolase A and C in the human central nervous system. AB - We have analyzed the distribution of aldolase A and C mRNAs and proteins in various areas of the human brain using Northern blot analyses and immunohistochemistry. Aldolase A mRNA expression was higher than aldolase C mRNA expression in all areas of the brain examined. Aldolase C mRNA expression was highest in the cerebellum. Aldolase C protein was present in well-delimited regions of the CNS, and was distributed in stripes in the Purkinje cell layer of the cerebellum, in the inferior olives and in the sensory neurons of the posterior horn of the spinal cord. The novel finding of aldolase C in well delimited cell compartments of the human cerebellum and in several other areas of the CNS lends weight to the hypothesis that this protein exerts other functions (e.g. sensory transmission) besides those characteristic of a glycolytic enzyme. PMID- 12626878 TI - Age-related change of neuropeptide Y-immunoreactive neurons in the rat anterior olfactory nucleus. AB - Neuropeptide Y (NPY) is located in the olfactory system, including the olfactory bulb, and is thought to be one of the main neurotransmitters for olfaction. Thus, we examined age-related changes of NPY-immunoreactive (IR) neurons in the rat anterior olfactory nucleus (AON) at various aging stages over a period of 2 years; postnatal months 1 (PM 1), PM 6, PM 12 and PM 24. NPY-IR neurons in the AON were present in the lateral and medial subdivisions at PM 1 and at PM 6 were distributed in all subdivisions of the AON. Prior to PM 12, the NPY-IR neurons showed a tendency to change from bipolar cells with short processes into multipolar cells with long processes. Moreover, the population of NPY-IR neurons and nerve fibers in the AON increased in proportion to age. In particular, the number of NPY-IR neurons increased about 6-fold between PM 1 and PM 3. At PM 24, the number of NPY-IR neurons was much smaller than that at PM 12 and somal size had decreased. It is therefore suggested that the dramatic increase in the number and size of the NPY-IR neurons between PM 1 and PM 3 may be associated with sexual maturation and that the decrease in the number and cell size of the NPY-IR neurons at PM 24 may underlie age-related changes in the olfactory process. PMID- 12626879 TI - Episodic exposure to ethanol during development differentially affects brainstem nuclei in the macaque. AB - Neuronal vulnerability to ethanol may be non-specific, i.e., vulnerability may be conferred by the developmental state of the population or by the site of derivation. To address these issues, the effect of developmental exposure to ethanol on three brainstem nuclei; the trigeminal motor (MoV), facial motor (MoVII) and medial superior olivary (MSO) nuclei was determined. MoVII and MSO are generated at the same time and from the same rhombomere, r4. MoV is generated earlier from r2. Macaca nemestrina were exposed to ethanol or a control solution one day per week for six or 24 weeks of gestation. Brainstems of the mature offspring were sectioned and stained. The number of neurons and volume of each nucleus were determined stereologically. Neuron number was lower in MoV and MSO following exposure to ethanol whereas MoVII appeared unaffected. No significant effects of ethanol exposure were seen on the volume and weight of the brainstem, or the volume of the individual nuclei. These findings show that ethanol differentially affects brainstem nuclei in a targeted, rather than non-specific, manner. Furthermore, they show that serious ethanol-induced neurological deficits can be present without gross morphological changes. PMID- 12626880 TI - Differential effects of intravitreal optic nerve and sciatic nerve grafts on the survival of retinal ganglion cells and the regeneration of their axons. AB - We have investigated the effects of intravitreal sciatic nerve (SN) and/or optic nerve (ON) grafts on the survival and the axonal regeneration of retinal ganglion cells (RGCs). Following transection of the ON, approximately 40% RGCs survived at 7 days post-axotomy (dpa). Results showed that the intravitreal ON graft significantly promoted the survival of RGCs at 7 dpa (39,063 vs 28,246). Intravitreal SN graft, however, did not rescue axotomized RGCs at 5, 7 or 14 dpa. Axotomized RGCs could be induced to regenerate axons along a segment of SN graft attached to the proximal stump of ON. On average, 608 axotomized RGCs were induced to regenerate axons along the attached SN graft. The presence of intravitreal SN graft promoted about 100% increase in the number of regenerating RGCs (1,227) relative to the control groups. The intravitreal ON graft, surprisingly, also induced about 100% more regenerating RGCs (1220) than in the control group. When SN and ON grafts were co-transplanted into the vitreous, about 200% more regenerating RGCs (1916) were observed than in the control group. These findings illustrated that the intravitreal ON graft rescued axotomized RGCs and enhanced the regeneration of retinal axons. This is the first report to show that ON promotes RGC axonal regeneration. The intravitreal SN graft did not rescue RGCs but promoted axonal regeneration. The differential effects of intravitreal ON and SN grafts on the survival and the RGC regeneration suggest that these might be two independently operating events. PMID- 12626881 TI - Pavlovian conditioning-specific increases of the Ca2+- and GTP-binding protein, calexcitin in identified Hermissenda visual cells. AB - Hermissenda CNS, immunolabeled for the memory protein calexcitin showed significant immunostaining over background in the B-photoreceptor cells of the eye. The degree of staining correlated positively with the number of Pavlovian training events experienced by the animals and the degree of Pavlovian conditioning induced. The training regime consisted of exposing animals to light (conditioned stimulus, CS) paired with orbital rotation (unconditioned stimulus, US). In animals that exhibited the conditioned response, calexcitin immunolabeling was more intense than was found for naive (unconditioned) animals or animals given the CS and US in random sequence. Animals exposed to lead (maintained in 1.2 ppm lead acetate) at a dosage known to impair learning in children, showed reduced learning and less intense calexcitin staining whether the CS and US were paired or given randomly. However, the levels were still higher than that of naive animals. Immuno-electron microscopy indicated that the labeling was predominantly within calcium sequestering organelles such as the endoplasmic reticulum, and to lesser extent within mitochondria, and photopigments. The calexcitin density after a short-term memory (STM) regime was the same whether measured 5 minutes after conditioning (when STM was evidenced by foot contraction) or 90 minutes later when no recall was detected. The staining density was also similar to the levels found 5 minutes after long-term memory (LTM) conditioning. However, the LTM regime produced a greater calexcitin intensity at 90 minutes when the memory had been consolidated. This learning specific increase in calexcitin is consistent with the previously implicated sequence of molecular events that are associated with progressively longer time domains of memory storage. PMID- 12626882 TI - Protease inhibitor-associated diabetes mellitus: a potential cause of morbidity and mortality. PMID- 12626883 TI - Optimization of ex vivo activation and expansion of macaque primary CD4-enriched peripheral blood mononuclear cells for use in anti-HIV immunotherapy and gene therapy strategies. AB - The rhesus macaque model is a useful experimental system to evaluate effects of T cell autotransfusion and gene therapies for HIV-1 infection and AIDS prior to a clinical trial. To obtain sufficient numbers of primary macaque CD4 T lymphocytes for this purpose, we examined the culture conditions that were needed to optimize ex vivo activation and expansion of macaque primary CD4-enriched peripheral blood mononuclear cells (PBMCs). In this report, we compared the effects of various stimulants on cell expansion, surface expression of CCR5 and CXCR4, and levels of transduction with a Moloney leukemia virus (MoLV) vector encoding the phenotypic selection marker truncated human nerve growth factor receptor (deltaNGFR) alone or with the human anti-HIV-1 tat intrabody sFvhutat2. The use of feeder cells strikingly increased the proliferation rate of macaque CD4-enriched PBMCs in vitro. In the presence of an irradiated rhesus macaque B-lymphoblastoid cell line (BLCL), the highest cell expansion over 21 days was achieved with cells activated by Con A (9648-fold), in turn, from high to low, phytohemagglutinin (PHA) (4855 fold), and anti-CD3/CD28-coated beads (2367-fold). Further studies showed that BLCL feeder cells were more effective than human PBMCs (hPBMCs) in promoting proliferation of macaque CD4-enriched PBMCs activated with Con A and anti CD3/CD28, respectively. The combined use of both BLCL and hPBMC feeder cells did not further increase cell expansion when compared with the use of BLCL cells alone. In addition, the addition of BLCL-conditioned medium (CM) and hPBMC-CM induced cell growth at a rate higher than did the culture medium alone but not as high as with feeder cells. Con A-activated macaque CD4-enriched PBMCs retained 88% of CXCR4 and 39% of CCR5 expression over 17 days compared with PHA-activated cells (50% for CXCR4, 16% for CCR5) and anti-CD3/CD28-activated cells (34% for CXCR4, 37% for CCR5). Finally, PHA, Con A, and CD3/CD28-coated beads supported comparable levels of MoLV transduction. The results should improve the utility of the rhesus macaque model for the testing of T-cell autotransfusion and gene therapies for HIV-1 infection/AIDS. PMID- 12626884 TI - Relative anti-HIV-1 efficacy of lamivudine and emtricitabine in vitro is dependent on cell type. AB - OBJECTIVE: To assess the relative in vitro potency of the antiviral agents emtricitabine (FTC), lamivudine (3TC), and zidovudine (ZDV) in peripheral blood mononuclear cells (PBMCs), monocyte-derived macrophages, and MT-4 cells infected with HIV-1. DESIGN In vitro evaluation of the test compounds against M-tropic or T-tropic laboratory strains of HIV-1 and against clinical HIV-1 isolates from antiretroviral therapy-naive subjects using PBMCs, monocyte-derived macrophages, and MT-4 cells. METHODS: Standard methods for assessing antiviral potency based on 50% inhibitory concentrations using propidium iodide staining of host cell DNA to assess cytopathic effects or measurement of HIV-1 reverse transcriptase activity to assess inhibition of viral replication. RESULTS: There were no significant differences in potency between 3TC and FTC in assays with HIV-1(IIIB) infected PBMCs or HIV-1(Ba-L) -infected monocyte-derived macrophages, which are primary cell types for HIV-1 infection in vivo. In agreement with earlier reports, FTC was approximately fourfold more active than 3TC in assays in the transformed T-cell line MT-4 infected with HIV-(1IIIB), whereas ZDV was more active than FTC. 3TC, FTC, and ZDV were equally active against a panel of eight primary HIV-1 isolates from antiretroviral-naive subjects in PBMCs. These results demonstrate the in vitro similarity of 3TC and FTC activity in primary cells. The variability in potency depending on cell types and viral strains underscores our observation that antiviral effects in vitro are not reliable predictors of in vivo clinical activity. PMID- 12626885 TI - Severe hepatotoxicity during combination antiretroviral treatment: incidence, liver histology, and outcome. AB - OBJECTIVES: To assess incidence, risk factors, histology, and outcome of severe hepatotoxicity (SH) during antiretroviral treatment (ART). METHODS: Seven hundred fifty-five HIV-seropositive patients consecutively prescribed new ART were selected. Liver function tests were assessed at baseline, after 1 month, and every 4 months thereafter. Liver biopsy was recommended in case of SH (i.e., increase in liver enzymes >/=10 times the upper limit of normal or 5 times baseline if markedly abnormal). RESULTS: Twenty-six cases of SH were observed with an incidence of 4.2% person-years. Liver failure (LF) was rarely seen (1.1 per 100 person-years). Liver damage was invariably observed in patients with chronic viral hepatitis. Liver histology showed exacerbation of viral hepatitis in all 16 patients for whom a liver biopsy was available at the time of SH. A direct correlation was found between alanine aminotransferase increase and increase in CD4 T-cell count in patients with SH (r = 0.53, p <.001). Death occurred during follow-up in 7 of 26 (27%) patients, all of whom showed LF and baseline CD4+ count less than 200 cells/mm(3) (7/7 patients = 100% vs. 8/19 patients without LF; p <.01). Relapse of SH was observed after ART was recommenced in 7 of 17 (41%) patients. Five of these 7 patients did not show further SH relapse after treatment with interferon. CONCLUSIONS: This study provides estimates of SH and LF in a large population-based setting where hepatitis C virus coinfection is highly prevalent and provides indications that liver damage may be caused by immune reconstitution and related exacerbation of viral hepatitis. A strict follow-up for hepatotoxicity is mandatory when ART is initiated in patients with <200 CD4+ T cells/mm(3). Antihepatitis pre- or comedication could be an effective preventive or curative measure. PMID- 12626886 TI - Real versus virtual phenotype to guide treatment in heavily pretreated patients: 48-week follow-up of the Genotipo-Fenotipo di Resistenza (GenPheRex) trial. AB - We compared viroimmunologic response after real phenotype (r-PHT) versus virtual phenotype (v-PHT) in patients failing highly active antiretroviral therapy (HAART). A total of 201 patients with >2 years of exposure, more than six experienced drugs, >1000 HIV RNA copies/mL, and on stable HAART for >6 months were randomized to the r-PHT or v-PHT arm. The primary end point was the proportion of HIV plasma viral load (pVL) <400 copies/mL. Secondary end points were absolute pVL change, proportion of pVL reduction >0.5 log(10) copies/mL, and absolute CD4 cell change. In the intention-to-treat-last observation carried forward analysis, study outcomes were not significantly different between arms over 48 weeks of follow-up: 20% and 24% pVL <400 copies/mL; 58% and 61% pVL reduction >0.5 log(10) copies/mL; -0.92 and -0.94(10) log copies/mL mean pVL decrease; and +41.6 and +94.4 cells/mm(3) mean absolute CD4 increase in the r-PHT and v-PHT arms, respectively. On-treatment analyses gave similar results. In the multivariate analysis of pVL <400 copies/mL, the following covariates were independent predictors at week 48: adherence (OR p= 0.25; p=.002), baseline CD4 (OR = 4.39; p=.007), intravenous drug use as risk factor for HIV acquisition (OR = 0.33; p=.024), and sensitivity score of the new regimens by biologic cut-offs (OR = 1.84; p=.029). Prescribed drugs for which patients were naive resulted in marginal prediction (OR = 1.93; p=.054). In conclusion, virologic and immunologic outcomes did not differ when r-PHT or v-PHT was used in this cohort of heavily pretreated patients. Several factors should be considered to take better advantage of resistance testing, including treatment history, clinical status, and patients' ability to adhere to treatment. PMID- 12626887 TI - A study of the immunology, virology, and safety of prednisone in HIV-1-infected subjects with CD4 cell counts of 200 to 700 mm(-3). AB - Adult Clinical Trials Group Study 349 examined the immunology, virology, and safety of 40 mg/d prednisone as an adjunct to antiretroviral therapy in 24 HIV infected subjects with >200 CD4+ T cells/mm in a randomized placebo-controlled trial. After 8 weeks, median lymphocyte and CD4+ cell numbers increased >40% above baseline values (p =.08). No effect was observed on markers of cell activation or apoptosis, although the proportion of CD28+ CD8+ T cells increased (p =.006). Prednisone inhibited monocyte TNFalpha production without affecting T cell responses to antigens or mitogens. Two subjects assigned to prednisone were subsequently found to have asymptomatic osteonecrosis of the hip. Many questions remain regarding the role of activation-induced sequestration and apoptosis as causes of progressive CD4+ T-cell loss in AIDS. The potential role of corticosteroids as tools to examine this question will be limited by concerns regarding their toxicity; however, further studies of other agents to limit cellular activation in AIDS are warranted. PMID- 12626888 TI - Evaluation of antiretroviral drug measurements by an interlaboratory quality control program. AB - Since 1999 an ongoing international interlaboratory quality control program has analyzed antiretroviral drugs in plasma. Results of the third round of this program are presented. Quality control samples were prepared by spiking drug-free plasma with varying concentrations of the currently available protease inhibitors and the nonnucleoside reverse transcriptase inhibitors efavirenz and nevirapine. Thirty-three laboratories participated in the program and were requested to analyze the quality control samples. Results were from 30 laboratories. Of all measurements, 82% were performed within 80%-120% accuracy limits. Only 3 laboratories performed all their measurements within these limits, and 12 participants reported at least 90% of their analyses within the acceptance range. Mean accuracy for low drug concentrations was worse than for medium and high concentrations. The percentage of satisfactory measurements for the 6 laboratories that participated for the third time in the program increased from 54% in the first round to 85% in the third round. The program revealed a large variability in the laboratories' ability to measure antiretroviral drugs accurately. This variability may have important implications for therapeutic drug monitoring of these drugs and for pharmacokinetic studies. Interlaboratory testing is useful to alert laboratories to previously undetected analytical problems. PMID- 12626889 TI - Prevalence of genotypic drug resistance among a cohort of HIV-infected newborns. AB - A retrospective, blinded study was conducted to examine the prevalence of antiretroviral drug resistance among a cohort of HIV-infected infants born in 1998 and 1999 in New York State. The earliest available HIV-positive specimen was tested. Most samples were from infants younger than 60 days of age. Genotype data were generated for the protease and reverse transcriptase genes of HIV-1 proviral DNA from 91 infected infants. Eleven infants (12.1%) had provirus with mutations associated with drug resistance, with all three classes of antiretroviral drugs represented. Two infants (2.2%) had mutations associated with resistance to two classes of antiretrovirals. Perinatal antiretroviral drug exposure was examined; it was not found to be significantly associated with the presence of resistance mutations. However, for those infants who had perinatal antiretroviral exposure and genotypic evidence of drug resistance to HIV, the mutations that were detected correlated with at least one antiretroviral from the perinatal period. The prevalence of genotypic drug resistance among this infant cohort is comparable with that found among recently infected adults. These results suggest that resistance testing should be strongly considered for perinatally infected infants, at the earliest possible time point, to avoid use of antiretroviral drugs to which the infant has preexisting resistance. PMID- 12626890 TI - Protease inhibitor use and the incidence of diabetes mellitus in a large cohort of HIV-infected women. AB - OBJECTIVE: To assess the association between protease inhibitor (PI) use and the incidence of diabetes mellitus (DM) among participants in the Women's Interagency HIV Study. DESIGN: Prospective multicenter cohort study. The diagnosis of DM was based on self-report at semiannual interviews conducted from 1994 to 1998. SETTING: Six inner-city clinical sites in the United States (Brooklyn, NY; Bronx, NY; Washington, DC; Chicago, IL; San Francisco, CA; and Los Angeles, CA). PARTICIPANTS: A total of 1785 nonpregnant women who had no history of prior DM. The women made up four groups: 1) PI users (n = 609, person-years [PY] at risk = 707); 2) reverse transcriptase inhibitor (RTI)-only users (n = 932, PY = 1486); 3) HIV-infected women reporting no antiretroviral therapy (ART) ever (n = 816, PY = 1480); and 4) HIV-uninfected women (n = 350, PY = 905). MAIN OUTCOMES: Incidence of DM and median body mass index (BMI) from 1995 to 1998 were compared among the four groups. RESULTS: Sixty-nine incident cases of DM occurred among 1785 women (1.5 cases per 100 PY; 95% CI: 1.2-1.9). The incidence of DM among PI users was 2.8 cases per 100 PY (2.8%) versus 1.2% among both RTI users and women on no ART (95% CI: 1.6-4.1 [PI]; 0.7-1.8 [RTI and no ART]; P = 0.01 for comparison of the PI group with the RTI group) and 1.4% among HIV-uninfected women (95% CI: 0.7-2.2, P = 0.06 for comparison with PI group). Weight gain was not associated with either PI or RTI use. Multivariate models identified PI use (hazard ratio [HR] = 2.90 [95% CI: 1.50-5.60]; P = 0.002), age (HR = 1.75 per 10 years [95% CI: 1.31-2.34]; P = 0.0002) and BMI as independent risk factors for DM. CONCLUSIONS: PI use was associated with a threefold increase in the risk of reporting incident DM. Routine screening for diabetes, particularly among older and heavier patients using PI therapy, is advisable. PMID- 12626891 TI - Short-term CD4 cell response after highly active antiretroviral therapy initiated at different times from seroconversion in 1,500 seroconverters. AB - The effect of HIV infection duration and CD4 cell count on short-term CD4 response was evaluated in treatment-naive seroconverters using logistic regression adjusted for CD4 count before highly active antiretroviral therapy (HAART) as well as for exposure category, age, sex, acute infection, and cohort. This association was also investigated in pretreated seroconverters, further adjusting for prior therapy. CD4 response (increase of >100 cells/microL at 6 months) was more likely if HAART was initiated in the first year following seroconversion (OR = 1.50 [95% CI: 1.07-2.10] compared with 2-5 years). There was no improvement in response from initiating HAART with CD4 count >350 cells/microL compared with 201 to 350 cells/microL. Below 200 cells/microL, however, the chance of a CD4 response appeared to be reduced (OR = 0.72 [95% CI: 0.40-1.28] for 0-200 cells/microL compared with 201-350 cells/microL, P = 0.26). Results were similar for pretreated individuals. Further, in pretreated individuals, a CD4 response was less likely if the CD4 nadir was lower than the pre-HAART CD4 count (OR = 0.18 [95% CI: 0.10-0.36] for >150 cells/microL difference between nadir and pre-HAART CD4 count vs. no difference, P < 0.001). Given the limitations of observational studies, particularly the inability to control for unmeasured confounders, these findings suggest that the initiation of HAART within the first year following seroconversion appears to improve short-term immunologic response. After that time, there is little to be gained in terms of short-term response from initiating HAART before reaching a CD4 count of 200 cells/microL. PMID- 12626892 TI - Assessing the magnitude of the HIV/AIDS epidemic in Burma. AB - OBJECTIVES: Estimates of the HIV/AIDS burden in Burma (Myanmar) are uncertain. Using data from the 1999 national HIV sentinel surveillance and available population data, we generated estimates of Burma's HIV burden in 1999. METHODOLOGY: The 1999 sentinel surveillance included women attending antenatal clinics, male military recruits, blood donors, injecting drug users, patients of sexually transmitted disease clinics, and sex workers. We used data for women attending antenatal clinics and male recruits aged 20-29 years to estimate HIV prevalence among women and men, respectively. Data points were merged to give five regional estimates of prevalence for men and women. Census figures were used to obtain national population estimates of the numbers of Burmese living with HIV infection, along with confidence intervals (CIs). RESULTS: HIV prevalence varied by region, with the lowest rates in the West, intermediate rates in the central region, and highest rates in the North, East, and South. The highest rates were in the East (Shan State), with female prevalence of 3.0% (95% CI, 1.9-4.5). The total number of infected women nationwide was 218,300 (95% CI, 159,400-277,100), and that of men was 468,700 (95% CI, 343,300-594,200). We estimated HIV prevalence of at least 3.46% (95% CI, 2.72-4.19) among adults aged 15-44 years; 5700 infants were born with HIV infection in 1999. DISCUSSION: Burma has a generalized epidemic of HIV-1 in reproductive age adults. We estimated that there were 687,000 (95% CI, 541,100-832,900) Burmese adults living with HIV infection in 1999, or about one of every 29 adult citizens. This estimate is higher than the UNAIDS estimate for the same year of 530,000 adults and children living with AIDS, or a population prevalence of about one in 50 adults. HIV prevention and care programs are urgently needed in Burma. PMID- 12626893 TI - Overcoming barriers to HIV testing: preferences for new strategies among clients of a needle exchange, a sexually transmitted disease clinic, and sex venues for men who have sex with men. AB - OBJECTIVE: To determine strategies to overcome barriers to HIV testing among persons at risk. METHODS: We developed a survey that elicited testing motivators, barriers, and preferences for new strategies among 460 participants at a needle exchange, three sex venues for men who have sex with men, and a sexually transmitted disease clinic. RESULTS: Barriers to testing included factors influenced by individual concern (fear and discrimination); by programs, policies, and laws (named reporting and inability to afford treatment); and by counseling and testing strategies (dislike of counseling, anxiety waiting for results, and venipuncture). The largest proportions of participants preferred rapid testing strategies, including clinic-based testing (27%) and home self testing (20%); roughly equal proportions preferred oral fluid testing (18%), urine testing (17%), and standard blood testing (17%). One percent preferred home specimen collection. Participants who had never tested before were significantly more likely to prefer home self-testing compared with other strategies. Blacks were significantly more likely to prefer urine testing. CONCLUSIONS: Strategies for improving acceptance of HIV counseling and testing include information about access to anonymous testing and early treatment. Expanding options for rapid testing, urine testing, and home self-testing; providing alternatives to venipuncture; making pretest counseling optional; and allowing telephone results disclosure may encourage more persons to learn their HIV status. PMID- 12626894 TI - Human T-cell lymphotropic virus type I (HTLV-I)-related clinical and laboratory findings for HTLV-I-infected blood donors. AB - Clinical and laboratory findings were examined for 111 human T-cell lymphotropic virus type I (HTLV-I)-infected blood donors. HTLV-I provirus loads in subjects with a family history of adult T-cell leukemia (ATL) or HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) tended to be higher than those in subjects without a family history of these conditions. There were 3 asymptomatic patients with ATL, 4 with a history of uveitis, 7 with hyperreflexia in the lower limbs, and 3 with urinary frequency in the night. The mean CD4 cell/CD8 cell ratio +/- SD was significantly lower (p<.0001) in subjects with hyperreflexia in the lower limbs (1.3 +/- 0.2) than in subjects without any clinical abnormalities (1.7 +/- 0.6), suggesting that subjects with hyperreflexia in the lower limbs already have some immunologic abnormalities. The concordance of HTLV-I infection between husband and wife was lower in this study than in a previous study. HTLV-I-related inflammatory symptoms were more frequent (p =.021, Fisher exact test; OR = 9.5; 95% CI, 1.7-53.5) in HTLV-I tax A-infected donors (3 [50%] of 6 donors) than in HTLV-I tax B-infected donors (10 [9.5%] of 105 donors), suggesting different risks of HTLV-I-related symptoms according to the virus genotype. PMID- 12626895 TI - Deleterious genetic influence of CX3CR1 genotypes on HIV-1 disease progression. AB - We previously reported that patients homozygous for a specific mutation (M280) in the chemokine receptor CX3CR1 progressed to AIDS more rapidly than those with other genotypes. This deleterious effect would predict that a cohort of prevalent patients would be depleted in M280 carriers, because these patients would have disappeared before recruitment. This hypothesis is confirmed in this new study based on the French SEROCO cohort showing that patients homozygous for the M280 allele were rare among the seroprevalent group. These results may explain the conflicting results published on the impact of CX3CR1 polymorphism in seroconverters. PMID- 12626896 TI - Molecular epidemiology of HIV-1 in Venezuela: high prevalence of HIV-1 subtype B and identification of a B/F recombinant infection. AB - The authors assessed HIV-1 variability in two distinct areas of Venezuela (the capital Caracas and Margarita Island) through the analysis of blood specimens and clinical and epidemiologic data of 72 persons. Proviral DNA was evaluated through heteroduplex mobility assay (HMA) based on the envelope region. Additionally, FOK I restriction enzyme digestion assay was performed in all subtype B ED31/33 amplified products to check the presence of the typical Brazilian subtype B GWGR variant. Sequencing and phylogenetic analysis for C2-V3 region of gp120 was performed in selected cases. The vast majority of samples were found to belong to subtype B, with a North American/European RFLP profile. An F subtype HIV-1 based on the region was identified for the first time in Venezuela. Genetic analyses of the protease and reverse transcriptase fragments of this sample depicted a recombinant B/F genetic profile. The discrimination capacity of HIV-1 subtypes using the primer set ED3/14-ED31/33 among the Caracas samples was found to be higher than for those from Margarita Island. The authors' results point to relevant differences between the samples of continental and Caribbean regions of Venezuela, requiring further evaluation of larger samples. PMID- 12626897 TI - Bacterial and mycobacterial meningitis in HIV-positive compared with HIV-negative patients in an internal medicine ward in Libreville, Gabon. PMID- 12626898 TI - Improved outcome of AIDS-related lymphoma in patients with virologic response to highly active antiretroviral therapy. PMID- 12626899 TI - Hepatitis C virus in patients with HIV infection and lipodystrophy. PMID- 12626900 TI - Unanswered questions about sexual transmission of HIV in Mwanza, Tanzania. PMID- 12626901 TI - Randomized trial of rubber band ligation vs. stapled hemorrhoidectomy for prolapsed piles. AB - PURPOSE: The introduction of stapled hemorrhoidectomy may replace local techniques such as rubber band ligation as a first-line treatment for Grade III and small Grade IV piles. We conducted a randomized trial to determine the role of rubber band ligation in the era of stapled hemorrhoidectomy. METHODS: Fifty five patients with Grade III or small Grade IV hemorrhoids were randomly allocated to either rubber band ligation or stapled hemorrhoidectomy. Patient demographics and procedure-related details were recorded. Follow-up was at two weeks and two and six months to assess complications, symptom relief, incontinence scores, quality of life, and patient satisfaction. RESULTS: Twenty five patients were randomly assigned to rubber band ligation and 30 to stapled hemorrhoidectomy. The groups were equally matched for age, gender, grade of piles, continence scores, and quality of life. Stapled hemorrhoidectomy was associated with increased pain and analgesia usage at both 2-week and 2-month follow-up (P < 0.001). Rubber band ligation and stapled hemorrhoidectomy were equally effective in controlling symptomatic prolapse, but rubber band ligation was associated with an increased incidence of recurrent bleeding (P = 0.002). There were 6 procedure-related complications in the stapled hemorrhoidectomy group compared with none in the rubber band ligation group (P = 0.027). There was no difference between the two groups in terms of continence scores, patient satisfaction, or quality of life. CONCLUSION: Stapled hemorrhoidectomy is associated with more pain and minor morbidity than rubber band ligation in the treatment of Grade III and small Grade IV piles. However, for those patients who do not want the risk of further intervention procedures, stapled hemorrhoidectomy offers the better chance of a symptomatic cure. PMID- 12626903 TI - A pathologic complete response to preoperative chemoradiation is associated with lower local recurrence and improved survival in rectal cancer patients treated by mesorectal excision. AB - PURPOSE: Preoperative chemoradiation reduces tumor size and nodal metastasis in patients with rectal cancer. Tumor downstaging has been associated with an increased probability of a sphincter-saving procedure and with improved local control. However, pathologic complete response to chemoradiation has not been correlated with local control and patient survival. We studied the prognostic value of pathologic complete response to preoperative chemoradiation in rectal cancer patients. METHODS: We have prospectively followed up 168 consecutive patients with ultrasound Stages II (46) and III (122) rectal cancer treated by preoperative chemoradiation followed by radical resection with mesorectal excision; 161 had a curative resection. Recurrence and survival were compared with tumor characteristics and pathologic complete response. Average follow-up was 37 months. RESULTS: Tumor downstaging occurred in 97 (58 percent) patients, including 21 (13 percent) patients who had a pathologic complete response. None of the clinical or pathologic variables was associated with pathologic complete response. The estimated 5-year rate of local recurrence was 5 percent; of distant metastasis, 14 percent. None of the patients with pathologic complete response has developed disease recurrence. We found no difference in survival among patients with pathologic Stages I, II, or III tumors. CONCLUSIONS: A pathologic complete response to preoperative chemoradiation is associated with improved local control and patient survival. For patients without pathologic complete response, the pathology stage does not have prognostic significance. PMID- 12626905 TI - Individual cutoff levels of carcinoembryonic antigen and CA 242 indicate recurrence of colorectal cancer with high sensitivity. AB - PURPOSE: This study was designed to identify the sensitivity and specificity associated with recurrent colorectal cancer, principally hepatic metastases, with individual cutoff levels of carcinoembryonic antigen; identify the corresponding data for CA 50 and CA 242; compare these findings with standard cutoff levels, clinical examinations, and patients' personal health assessments; and identify the time between increasing serum levels and detection of actual relapses, particularly hepatic metastases. METHODS: A prospective study was performed that included 132 patients with colorectal cancer who underwent surgery with curative intent and who were followed up for a minimum of 5 years. Serum samples were obtained the first two years after surgery. Marker serum levels were analyzed with the commercial Delfia test kit. RESULTS: During the sampling period, carcinoembryonic antigen levels were very high or above the individual cutoff level in 21 of 24 cases with recurrent disease. All eight hepatic metastases detected during the sampling period were indicated by carcinoembryonic antigen 0 to 19 months earlier. In the 3rd postoperative year, nine patients with recurrence were identified, and in eight of them, carcinoembryonic antigen rose above the individual cutoff level 23 months before diagnosis. The overall sensitivity and specificity associated with recurrent disease with individual cutoff levels were 79 and 64 percent for carcinoembryonic antigen, 46 and 77 percent for CA 50, and 63 and 76 percent for CA 242, respectively, which was higher than for any other method compared. CONCLUSIONS: With individual cutoff levels, both carcinoembryonic antigen and CA 242 had high sensitivity and acceptable specificity for indicating recurrent colorectal cancer. Because most (29/33) patients with recurrent disease identified within 3 years after surgery and all 8 with hepatic metastases identified during the sampling period were indicated by carcinoembryonic antigen with the individual cutoff, that method was established as the best for postoperative surveillance. PMID- 12626904 TI - Early-age-at-onset colorectal cancer and microsatellite instability as markers of hereditary nonpolyposis colorectal cancer. AB - PURPOSE: Early-age-at-onset colorectal cancer and microsatellite instability are characteristic features of hereditary nonpolyposis colorectal cancer. Our aim was therefore to investigate whether these features might be useful markers in screening for hereditary nonpolyposis colorectal cancer and mismatch repair gene mutations. METHODS: From 1,132 consecutive patients who underwent surgery for colorectal cancer at our department between 1980 and 1999, we selected all patients 40 years of age or younger (study group, n = 59) and a subset of patients 40 years of age or older (control group, n = 60) who were matched for gender and pathologic TNM stage. Patients for whom a complete family cancer history or microsatellite status was unavailable were excluded from the study. Family cancer histories, retrieved from archival charts, were reassessed. Microsatellite status was investigated with the five microsatellites from the Bethesda recommended panel (BAT-26, BAT-25, D2S123, D5S346, and D17S250). On the basis of the number of altered microsatellites (> or = 2, 1, or 0), tumors were considered as having high or low instability or microsatellite stability, respectively. Mutation analysis for MLH1 and MSH2 genes was performed only in cases of high instability. DNA was investigated for mutations by single-strand conformational polymorphism and sequencing analysis. RESULTS: Data from 95 patients (study group: n = 37, 18 males, mean age 35 years; control group: n = 58, 29 males, mean age 62 years) were available for analysis. Four patients (study group, n = 3; control group, n = 1) fulfilled the Amsterdam II criteria for hereditary nonpolyposis colorectal cancer. Of the 37 study group tumors, 12 (32.4 percent) showed high-frequency microsatellite instability, and 25 had microsatellite stability, whereas among the 58 control group tumors, 4 (7 percent) showed high-frequency microsatellite instability, and 54 had microsatellite stability (P < 0.002). Mismatch repair gene mutation analysis was performed in 12 cases (study group, n = 7; control group, n = 5). We found four mutations (MSH2 119delG, MLH1 ex9 684insT, MSH2 Gln239Stop, and MLH1 del0.8 Kb) in the study group patients and none in the control group. Of four hereditary nonpolyposis colorectal cancer patients who underwent mismatch repair gene mutation analysis, one had a mutation. CONCLUSIONS: Early-age-at-onset colorectal cancer is significantly correlated with high-frequency microsatellite instability tumor status and is a useful criterion to identify hereditary nonpolyposis colorectal cancer patients. Moreover, when used in association with high frequency microsatellite instability status, it is effective in selecting patients for mismatch repair gene mutation analysis. PMID- 12626906 TI - Randomized, controlled trial of carbon dioxide insufflation during colonoscopy. AB - INTRODUCTION: Insufflation of air is a cause of discomfort during and after colonoscopy. Although this can be minimized by good technique, the use of carbon dioxide insufflation may provide further benefits. Carbon dioxide is rapidly absorbed and excreted through the lungs. We hypothesized that carbon dioxide would alleviate post-colonoscopy discomfort. METHODS: After they had provided informed consent, patients presenting for colonoscopy were randomized into two groups: those in whom air was used for colonoscopy and those in whom carbon dioxide was used. Pain during and ten minutes after colonoscopy was measured on a ten-point analog scale. Data are mean and 95 percent confidence limits. RESULTS: There were 124 patients in the air group and 123 in the carbon dioxide group. Age, body mass index, indication, diagnosis, and number of procedures were similar for the two groups. There were no differences between the groups in the amounts of sedation or analgesia used, the percentage of examinations that were complete (air, 98.4 percent; carbon dioxide, 95.2 percent), or patient satisfaction (on a scale of 1 to 10: air, 9.4; carbon dioxide, 9.5). Although there were more females in the carbon dioxide group (69 vs. 51), hysterectomy rates were the same. Pain scores (mean +/- 95 percent confidence interval; scale of 1 to 10) immediately after the examination were 4.3 +/- 0.3 for air and 3.6 +/ 0.3 for carbon dioxide (no significant difference). Pain scores 10 minutes later were 2.1 +/- 0.2 for air and 0.9 +/- 0.2 for carbon dioxide (P < 0.05, Student's t-test). CONCLUSION: Because there was significantly less abdominal pain ten minutes after colonoscopy in the group in whom carbon dioxide was used, carbon dioxide should be considered as an insufflating gas for colonoscopy. PMID- 12626907 TI - Analysis of K-ras codon 12 mutation in flat and nodular variants of serrated adenoma in the colon. AB - PURPOSE: The developmental process of serrated adenomas is obscure, and the importance of genetic alterations has not been elucidated clearly. The possibility that the developmental process and genetic alterations of serrated adenomas could differ from those of ordinary tubular adenomas was explored in this work. METHODS: Serrated adenomas were obtained by endoscopic resection (n = 57) and divided into two groups: flat (n = 10) and nodular (n = 47). Mutation of the K-ras gene was analyzed by enriched polymerase chain reaction-enzyme-linked mini-sequence assay, which can detect not only the presence of a mutation but also the mutation type of K-ras codon 12 with high sensitivity. Methylation specific polymerase chain reaction was performed with specific primers for the DNA repair gene O6-methylguanine-DNA methyltransferase. RESULTS: Serrated adenomas located in the rectum were more likely to have a K-ras mutation (9/12, 75 percent), whereas serrated adenomas of the flat type were less likely to have one (1/10, 10 percent). Furthermore, nodular serrated adenomas that occurred in the rectum possessed a high frequency of K-ras gene codon 12 point mutation (8/10, 80 percent) despite an overall frequency of 46.8 percent (22/47). A mutation of the K-ras codon 12 gene was detected in 23 (40.4 percent) of 57 serrated adenomas. Three types of point mutations of codon 12 were detected, with the mutation of GAT being observed most frequently. CONCLUSIONS: This study shows that development of nodular serrated adenomas may depend on the mutation of the K ras codon 12 gene, whereas development of flat serrated adenomas may not. Additionally, serrated adenomas that occur in the rectum are closely related to the mutation of the K-ras codon 12 gene. K-ras mutations in serrated adenomas may be unaffected by the epigenetic silencing of O6-methylguanine-DNA methyltransferase by promoter hypermethylation. PMID- 12626909 TI - Endoscopic removal of large colorectal polyps: prevention of unnecessary surgery? AB - PURPOSE: Because of the potential risk of malignancy and technical difficulties in achieving complete removal, large colorectal polyps represent a special problem for the endoscopist. The aim of this study was to evaluate the capabilities and risks of endoscopy in complete removal of large colorectal polyps. METHODS: Endoscopic polypectomy of 186 colorectal polyps larger than 3 cm in diameter (range, 3-13 cm) was performed; 141 were sessile and 45 pedunculated. Most of the polyps were located in the rectum (n = 88), sigmoid (n = 63), and cecum (n = 9). The remaining adenomas were situated in other parts of the colon. Sessile polyps were removed using the piecemeal technique. RESULTS: Histology results showed an adenoma in 167 cases, and invasive carcinoma was present in the adenoma in 19 patients. Of the adenomas, 29 were tubulous, 118 tubulovillous, and 20 villous; adenoma with severe dysplasia was found in 49 cases. Complete endoscopic removal was achieved in all sessile and pedunculated polyps. None of the patients with invasive carcinoma who underwent surgical resection (n = 10) had any evidence of tumor in the resected specimen. Bleeding occurred in 4 patients after polypectomy (2 percent). Perforation occurred in 1 patient (0.5 percent), who had an invasive carcinoma of the cecum. There was no procedure related mortality. During a mean follow-up period of 40 (range, 3-87) months, 6 patients presented with recurrence of a benign adenoma (3 percent), which was treated endoscopically, and 1 patient presented with a recurrent invasive carcinoma, which was treated surgically. CONCLUSIONS: Endoscopic polypectomy is a safe and effective method of treating large colorectal polyps. PMID- 12626908 TI - Prognostic significance of lateral lymph node micrometastases in lower rectal cancer: an immunohistochemical study with CAM5.2. AB - PURPOSE: The aims of this study were to determine the rate of lymph node micrometastases and to evaluate their prognostic value in lateral lymph nodes in lower rectal cancer at or below the peritoneal reflection. METHODS: A retrospective analysis was made of 892 lymph nodes from 66 consecutive patients who had undergone radical resection with lateral lymph node dissection. These lymph nodes were examined immunohistochemically with an antibody against cytokeratins 7 and 8, CAM5.2. RESULTS: Routine hematoxylin-eosin staining revealed 9 patients with positive lateral lymph nodes that were stained consistently with CAM5.2. Among 57 patients in whom lateral lymph node metastases were not detected by hematoxylin-eosin staining, cytokeratin staining was positive in 19 nodes (2.7 percent) from 11 patients (19.3 percent). These 11 patients with micrometastases in lateral nodes showed a significantly high recurrence rate (P = 0.048) and worse overall survival (P = 0.01) than the 46 patients without lateral node metastases. The recurrence rate and overall survival of patients with micrometastases did not differ significantly from those of patients with positive lateral nodes with hematoxylin-eosin staining. Local recurrence developed in 6 of 66 patients, but neither the presence nor the absence of micrometastases in lateral nodes influenced the local recurrence rate. CONCLUSION: The presence of nodal micrometastases leads to a poor prognosis. The survival of patients with micrometastases was not different from that of patients with overt metastases. Therefore, patients with cytokeratin-positive cells in lateral lymph nodes should be regarded as having overt metastases. PMID- 12626910 TI - Virtual reality: new method of teaching anorectal and pelvic floor anatomy. AB - PURPOSE: A clear understanding of the intricate spatial relationships among the structures of the pelvic floor, rectum, and anal canal is essential for the treatment of numerous pathologic conditions. Virtual-reality technology allows improved visualization of three-dimensional structures over conventional media because it supports stereoscopic-vision, viewer-centered perspective, large angles of view, and interactivity. We describe a novel virtual reality-based model designed to teach anorectal and pelvic floor anatomy, pathology, and surgery. METHODS: A static physical model depicting the pelvic floor and anorectum was created and digitized at 1-mm intervals in a CT scanner. Multiple software programs were used along with endoscopic images to generate a realistic interactive computer model, which was designed to be viewed on a networked, interactive, virtual-reality display (CAVE or ImmersaDesk). A standard examination of ten basic anorectal and pelvic floor anatomy questions was administered to third-year (n = 6) and fourth-year (n = 7) surgical residents. A workshop using the Virtual Pelvic Floor Model was then given, and the standard examination was readministered so that it was possible to evaluate the effectiveness of the Digital Pelvic Floor Model as an educational instrument. RESULTS: Training on the Virtual Pelvic Floor Model produced substantial improvements in the overall average test scores for the two groups, with an overall increase of 41 percent (P = 0.001) and 21 percent (P = 0.0007) for third year and fourth-year residents, respectively. Resident evaluations after the workshop also confirmed the effectiveness of understanding pelvic anatomy using the Virtual Pelvic Floor Model. CONCLUSION: This model provides an innovative interactive educational framework that allows educators to overcome some of the barriers to teaching surgical and endoscopic principles based on understanding highly complex three-dimensional anatomy. Using this collaborative, shared virtual-reality environment, teachers and students can interact from locations world-wide to manipulate the components of this model to achieve the educational goals of this project along with the potential for virtual surgery. PMID- 12626911 TI - Open vs. laparoscopic surgery for rectal prolapse: a case-controlled study assessing short-term outcome. AB - PURPOSE: This study was undertaken to evaluate the efficacy and safety of laparoscopic repair for rectal prolapse. METHODS: A case-control study was undertaken. The case group consisted of a consecutive series of patients who underwent laparoscopic repair for rectal prolapse between February 1993 and June 2000. The control group underwent open prolapse repair between October 1987 and January 2000. RESULTS: There were 53 patients in each group. The groups were matched according to operation type, gender, and age. Median operative time was longer in the case group than in the control group (resection rectopexy 210 vs. 117 minutes, rectopexy 127.5 vs. 72 minutes, respectively). Median postoperative hospital stay was shorter in the case group than in the control group (resection rectopexy 5 vs. 7 days, rectopexy 4.5 vs. 7 days, respectively). Median intraoperative bleeding was minor in the case group (resection rectopexy 35 vs. 300 ml, rectopexy 15 vs. 100 ml, respectively). Mortality (0 vs. 4 percent), complications (23 vs. 30 percent), late complications (4 vs. 13 percent), and the rate of recurrent prolapse (6 vs. 13 percent) did not differ significantly between the groups. CONCLUSIONS: Laparoscopic repair for rectal prolapse is technically feasible and can be performed with mortality and morbidity rates comparable to those of the conventional technique. The main advantages of the laparoscopic approach appear to be a shorter hospital stay and lessened intraoperative blood loss. Recurrence rate is not increased in the short term. PMID- 12626912 TI - Botulinum toxin as second-line therapy for chronic anal fissure failing 0.2 percent glyceryl trinitrate. AB - PURPOSE: Glyceryl trinitrate paste is used by many as first-line therapy for chronic anal fissure but heals only approximately 50 to 60 percent of fissures. We use botulinum toxin as second-line therapy after failed glyceryl trinitrate and aimed to evaluate efficacy, side effects, and patient preference. METHODS: A prospective, nonrandomized, open-label study of patients with chronic anal fissure failing a course of glyceryl trinitrate treated with 20 units of botulinum toxin A injected into the internal sphincter was conducted. Symptomatic relief, visual healing of fissures, side effects, and patient preference were assessed at 8-week follow-up. RESULTS: Forty patients underwent botulinum toxin treatment. Twenty-nine patients (73 percent) overall were improved symptomatically and avoided surgery. Seventeen fissures (43 percent) were healed, whereas 23 fissures (57 percent) remained unhealed. Of the unhealed fissures, 5 (12 percent) were asymptomatic, 7 (18 percent) were symptomatically much improved, and 11 (27 percent) were no better symptomatically and came to surgery. Discomfort associated with injection was minimal. Of 34 patients undergoing botulinum toxin injection in the clinic, 24 (71 percent) preferred botulinum toxin, 7 glyceryl trinitrate (20 percent; difference = 51 percent; 95 percent confidence interval = 31-71 percent), and 9 percent were undecided. Transient minor incontinence symptoms were noted in 7 patients (18 percent). CONCLUSIONS: Second-line botulinum toxin injection improves symptoms in approximately three quarters of patients after failed primary glyceryl trinitrate therapy and at least in the short term avoids surgical sphincterotomy. Botulinum toxin heals approximately one-half of these fissures. Discomfort and side effects were minimal. A policy of first-line glyceryl trinitrate/second-line botulinum toxin will avoid sphincterotomy in 85 to 90 percent. Higher rates of healing may be achieved by giving botulinum toxin as first-line therapy, or addressing the chronic fibrotic nature of the fissure. PMID- 12626913 TI - Preoperative immune status determines anal condyloma recurrence after surgical excision. AB - PURPOSE: Immunosuppression is an important risk factor for anal condyloma progression; however, its effects on postoperative outcomes remain unclear. In the present study, we investigated risk factors and recurrence rates in immunocompromised patients requiring surgery for medically intractable anal condyloma. METHODS: A retrospective review was performed on 63 consecutive patients who underwent surgical intervention for medically intractable anal condyloma at a single institution. Patient cohorts included immunosuppressed patients (e.g, HIV-seropositive, leukemia, idiopathic lymphopenic syndrome, or transplant patients; n = 45) and immunocompetent patients (n = 18). Recurrence rates and time to recurrence after surgery were determined for both groups. Preoperative CD4 count and viral load within the HIV subpopulation were also determined. The majority of the patients were male (90 percent), with a median age of 36 years. RESULTS: Anal condyloma recurred in 66 percent of the immunosuppressed patients compared with 27 percent of the immunocompetent group. Recurrence time expressed in months was shorter in immunosuppressed patients than in immunocompetent patients (6.8 +/- 1.5 vs. 15 +/- 5.7 months; P< 0.05). In the subpopulation of HIV-seropositive patients, no association was found between recurrence rates and viral loads; however, CD4 counts were significantly lower in those who had recurrence than in those who did not (226 +/- 31.7 vs. 401 +/- 51.2 cells/microl; P< 0.05). CONCLUSIONS: After surgery, anal condyloma in immunosuppressed patients recurs significantly more often and within a shorter period of time than in patients with a competent immune system. Additionally, in HIV-seropositive patients, CD4 counts should be maximized to prevent early recurrence of anal condyloma. PMID- 12626914 TI - Morphology of ileal and jejunal pouches used as rectal substitutes. AB - PURPOSE: Our aim was to determine whether the mucosa of a canine jejunal pouch used as a rectal substitute after proctocolectomy retains its morphologic features better than the mucosa of a canine ileal pouch so used. METHODS: Among ten dogs that underwent proctocolectomy, five had a jejunal pouch-distal rectal anastomosis and five an ileal pouch-distal rectal anastomosis. After six months, the animals were killed, the intestinal pouches and portions of unaltered distal ileum were removed, and a blinded, mucosal morphometric analysis was performed. RESULTS: The mucosa of the jejunal pouches had an overall thickness (mean +/- standard deviation, anterior and posterior walls, 1,300 +/- 140 microm), villous height (286 +/- 46 microm), and crypt depth (790 +/- 77 microm) greater than that of the ileal pouches (920 +/- 170, 208 +/- 47, and 530 +/- 130 microm, respectively; P < 0.05). Moreover, the mucosal thickness of the jejunal pouches was similar to that of the distal ileum proximal to the pouch (1,200 +/- 200 microm; P > 0.05), whereas the mucosal thickness of the ileal pouch was thinner (P < 0.05). CONCLUSIONS: The jejunal mucosa retains its major morphometric features when the jejunum is used as a rectal substitute after proctocolectomy. In contrast, the ileal mucosa atrophies when the ileum is so used. PMID- 12626915 TI - Anal fistula: Levovist-enhanced endoanal ultrasound: a pilot study. AB - PURPOSE: The aim of the study was to investigate the usefulness of the contrast agent Levovist in ultrasound assessment of anal fistula. METHODS: Fifteen patients (11 females, mean age 46) with a diagnosis of anal fistula were assessed by physical examination, conventional ultrasound, Levovist-enhanced ultrasound, and surgery. Levovist was injected a cannula into the fistula. The results of physical examination, conventional ultrasound and Levovist-enhanced ultrasound were compared with surgical findings as criterion standard. RESULTS: At physical examination, three intersphincteric fistulas and two sinuses were diagnosed. Using conventional ultrasound, five intersphincteric and five transsphincteric fistulas were found; four fistulas and one sinus were not detected. Levovist enhanced ultrasound revealed one sinus, five intersphincteric, seven transsphincteric, and one extrasphincteric fistulas; only one fistula was not detected. At surgery, three intersphincteric, seven transsphincteric, and two sinuses were found; however, the extrasphincteric fistula detected by Levovist was missed. Compared with physical examination, Levovist-enhanced ultrasound and surgery were significantly favorable in the diagnosis of anal fistula (P < 0.05 in chi-squared test and Fisher's exact probability test). The concordance rate of surgery with conventional ultrasound was 69 percent (9/13) and with Levovist enhanced ultrasound was 77 percent (10/13). However, because the extrasphincteric fistula was missed at surgery, the accuracy of Levovist-enhanced ultrasound was in fact 85 percent (11/13) if surgical finding was not used as the standard. The internal opening was detected at physical examination in 2 patients (13 percent), with conventional ultrasound in 4 patients (27 percent), with Levovist-enhanced ultrasound in 9 patients (60 percent) and during surgery in 11 patients (85 percent). Consistently, Levovist-enhanced ultrasound and surgery were significantly better than physical examination in the diagnosis of internal opening (P < 0.05). One secondary extension and two sphincter defects were detected by both types of ultrasound. The extension was not confirmed during surgery. No patients developed recurrence or nonhealing of wound. One patient developed incontinence to flatus and one developed a perianal hematoma. CONCLUSION: Levovist-enhanced ultrasound is better at assessing anal fistula than physical examination and conventional ultrasound. However, a future trial comparing Levovist, hydrogen peroxide, and magnetic resonance imaging is needed to establish which is the most cost-effective preoperative imaging technique to use. PMID- 12626916 TI - Determinants of recurrence after sigmoid resection for uncomplicated diverticulitis. AB - PURPOSE: This study aimed to evaluate the impact of surgery-associated variables on recurrence rates after sigmoid resection for diverticulitis. METHODS: Patients who underwent elective sigmoid resection for uncomplicated diverticulitis between 1992 and 2000 at two tertiary referral centers were followed up for recurrent disease as the primary end point. Recurrence after surgery was defined as left lower quadrant pain, fever, and leukocytosis, with consistent CT and/or contrast enema findings on admission and after six weeks. A logistic regression of the following variables was undertaken: patient demographics, duration of preoperative symptoms, previous admissions and abdominal surgery, surgical access (laparoscopic or open), postoperative complications, splenic flexure mobilization, anastomotic technique (handsewn or stapled), specimen length, inflammation at proximal resection margin, and anastomotic level (colosigmoid or colorectal). The last three variables were defined by the pathologist. Anastomosis level was based on muscle layer configuration (taeniae coli) at the distal resection margin. RESULTS: Two hundred thirty-six patients (105 females) with a mean age of 60.4 (standard deviation, +/- 10) years were available for follow-up at 67 +/- 30 (range, 11-130) months. The median duration of preoperative symptoms was 18 (range, 12-120) months. All but one (99 percent) patient had at least one admission before surgery. One hundred forty (59 percent) and 96 (41 percent) patients underwent laparoscopic or open resection, respectively. The conversion rate was 13 percent (18 patients) in the former group and the 30-day complication rate was 23 percent, with 0.4 percent 30-day mortality and a 2.1 percent reoperation rate. The splenic flexure was mobilized in 109 patients (47 percent). Anastomoses were fashioned by stapler in 171 patients (73 percent) and were to the rectum in 143 patients (72 percent). Specimen length was 17.9 +/- 5.9 (range, 9-47) cm with inflammation at the proximal margin in 30 patients (14 percent). Twelve (5 percent) patients developed a recurrence at a mean of 78 +/- 25 (range, 34-109) months with reoperation in one (0.4 percent). The level of anastomosis was the only predictor of recurrence in regression analysis (P = 0.033). Patients with colosigmoid anastomosis had a four times higher risk of having a recurrence compared with patients with colorectal anastomosis (odds ratio, 95 percent confidence interval = 1.12, 14.96). CONCLUSION: Colorectal (rather than colosigmoid) anastomosis was the single predictor of lower recurrence rates after elective sigmoid resection for uncomplicated diverticulitis. PMID- 12626917 TI - Optimizing the outcome for patients with rectal cancer. AB - INTRODUCTION: Historically, rectal cancer with transmural spread and/or lymph node involvement has presented a major challenge to surgeons, with a variable and often high risk of local recurrence and poor survival outcomes. In recent years a large amount of literature has focused attention on the importance of surgical technique, tumor staging, and the optimal integration of CT and radiation therapy. METHODS: This article reviews the clinical trials that have defined the current approach to rectal cancer, the controversies regarding what should be considered the standard of care, and the ongoing clinical studies that will resolve some of these issues. RESULTS: The preoperative staging of rectal cancer can be improved with the use of endorectal ultrasound and (where available) magnetic resonance imaging. Careful pathologic analysis, particularly of the radial margin, provides important prognostic information that enables better allocation of postoperative care. Although both radiation therapy and CT have a proven role in adjuvant therapy, the interpretation of many studies is confounded by unacceptably poor outcomes in the control arm, and in older studies the use of inferior chemotherapy and radiation therapy techniques. Ongoing studies will better define the optimal combination and timing of chemotherapy and radiation therapy, with respect to both toxicity and survival endpoints. CONCLUSIONS: A combined modality approach to rectal cancer, integrating the colon and rectal surgeon, pathologist, medical oncologist, and radiation oncologist, is necessary to achieve optimal outcomes. The achievements to date and the ongoing vigorous debates regarding standard care continue to highlight the importance of quality ongoing research in a rapidly changing clinical environment. PMID- 12626918 TI - Nonsteroidal anti-inflammatory drug-induced colonic stenosis: an unusual cause of a right-sided colonic mass: report of a case. PMID- 12626919 TI - Initial presentation of Crohn's disease in pregnancy: report of a case. AB - PURPOSE: This study evaluates the current morbidity and mortality of Crohn's disease presenting for the first time in pregnancy. METHODS: A review of the English-language literature was performed to collect all reported cases of Crohn's disease presenting in pregnancy. RESULTS: This review demonstrates a maternal mortality of 4 percent and morbidity of 40 percent and a fetal mortality of 38 percent, with 24 percent normal outcome of pregnancy. CONCLUSIONS: This study shows improved maternal and fetal outcome compared with earlier data. PMID- 12626920 TI - Presacral neuroendocrine carcinoma developed in a tailgut cyst: report of a case. AB - We report a case of retrorectal neuroendocrine carcinoma in a 68-year-old male. This malignant tumor was associated with cystic structures with a variety of epithelial linings, including squamous, ciliated, columnar, and transitional, consistent with a tailgut cyst. To the best of our knowledge this is only the second such tumor reported in the literature. PMID- 12626921 TI - Minimally invasive sacral neuromodulation implant technique: modifications to the conventional procedure. AB - PURPOSE: Sacral neuromodulation is a novel treatment for selected urinary and bowel dysfunctions. A new method is described for electrode implantation, the "minimally invasive sacral neuromodulation implant technique." METHODS: After the percutaneous nerve evaluation test, a small longitudinal incision (3 cm) is made, and a catheter cannula segment is inserted through the sacral foramen beside an insulated needle. The electrode is introduced into the catheter cannula, which is then removed and fixed to the sacrum by means of small anchors. The proximal part of the lead is tunneled into the subcutaneous tissue, reaching the pocket made to accommodate the neurostimulator. RESULTS: This procedure was performed in ten patients (5 male; mean age, 50.4 years). In four patients a single electrode was implanted, and in six patients two electrodes were implanted. The minimally invasive technique was significantly faster, saving a mean time of 20 minutes for each electrode. The incision made directly on the sacral foramen was significantly reduced (3 vs. 12 cm), avoiding the wide, blunt dissection of subcutaneous fat tissue. Application of the catheter cannula allowed the electrode to be introduced easily and correctly. The electrode anchors never failed: no cases of lead displacement or suboptimal position of the electrode occurred. A unilateral, sterile subcutaneous seroma occurred in one of the ten patients. CONCLUSIONS: The minimally invasive sacral neuromodulation implant technique seems to be a safe procedure-making sacral neuromodulation implant easier, faster, and safer, in as much as complications could be potentially reduced. PMID- 12626922 TI - Cecoscopy can be dangerous in patients with carcinoma of the cecum. PMID- 12626924 TI - Safety of botulinum neurotoxin treatment in patients with chronic anal fissure. PMID- 12626925 TI - A prospective study on the implications of a base deficit during fluid resuscitation. AB - An excessive base deficit (BD) and elevated serum lactate are increasingly recognized as important markers of a malperfusion state during the resuscitation of thermally injured patients. In a previous retrospective study, we found that patients with a BD less than -6 mmol/l during fluid resuscitation developed more severe systemic inflammatory response syndrome (SIRS), more frequent acute respiratory distress syndrome (ARDS), and more severe multiple organ dysfunction syndrome (MODS). The object of this study was to reexamine prospectively the relationship between the BD during fluid resuscitation and the subsequent development of SIRS, ARDS, and MODS by undertaking a prospective observational study of a cohort of consecutive burn patients. Analysis was completed on 38 patients with a mean age of 39 +/- 17 years and a mean %TBSA burn of 36 +/- 15%. The mean BD in the first 24 hours was less than -6 mmol/l in five patients (BD24 < -6 group), and was greater than -6 mmol/L in 33 patients (BD24 > -6 group). Patients in both groups were resuscitated to nearly identical endpoints of urinary output (1.2 ml/kg/hr in the BD24 < -6 group vs 1.3 ml/kg/hr in the BD24 > -6 group). Patients in the BD24 < -6 group had a trend toward a greater number of SIRS signs on the first postburn day, had a significantly higher incidence of ARDS (P =.02), and had significantly more severe MODS (P <.001) than patients in the BD24 > -6 group. The results concur with those of our previous retrospective study. Despite resuscitation to an acceptable urinary output, some burn patients develop a more extreme BD and go on to experience more severe organ dysfunction than do patients who do not generate a BD. The effect of specific correction of the BD during fluid resuscitation is not known at this time. PMID- 12626926 TI - Trash and brush burning: an underappreciated mechanism of thermal injury in a rural community. AB - Open-fire injuries associated with burning trash or brush result in a significant number of burn admissions annually. Trash- and brush-burning injuries are more prevalent in rural environments where open burning is an acceptable practice of waste disposal. These injuries are not "unavoidable accidents" but follow a predictable injury pattern. Understanding this injury pattern is paramount to effective prevention. The purpose of this work is to describe the epidemiology of burn injuries associated with burning trash or brush. Charts were retrospectively reviewed for all trash and brush burning injuries from July 1989 to December 2000. One hundred ninety-four injuries were identified, accounting for one fifth of all flame admissions for the study period. Persons most at risk for these burn injuries were males between the ages of 25 to 45, followed by males ages 16 to 24. Accelerant use was identified in over 80% of these injuries. The median cost for admission was $9,363.00 US dollars. Cost was significantly related to age, percent body surface area burned, and operative intervention. Brush and trash burning can lead to costly injuries especially when accelerants are used. Additional research is needed to explore the cultural and social influences associated with these injuries to target effective prevention. PMID- 12626927 TI - OT/PT forum. PMID- 12626928 TI - Computerized manufacturing of transparent face masks for the treatment of facial scarring. AB - In this work an integrated system of computer-aided design and computer-aided manufacturing specifically designed to fabricate affordable transparent face masks is discussed. FaceScan, a custom-designed software system, integrates shape capture, mask design, and pattern fabrication. The software controls a linear scan noncontact laser imager for facial topography acquisition and a milling machine for pattern fabrication. Compared with conventional methods of mask fabrication, this system is faster, more accurate, and less stressful for the patient and allows for greater control of the finished product. Masks for two subjects have been successfully fabricated using this system. PMID- 12626929 TI - Use of a modified tilt table for preambulation strength training as an adjunct to burn rehabilitation: a case series. AB - Because of the system-wide complications that arise with prolonged bed rest, early mobilization plays a critical role in the recovery process, especially in the patient with significant burn injury. Unfortunately, early mobilization of patients with severe burns can be a difficult and uncontrolled task and often requires several people to lift a patient to a standing position. This article describes the use of a modified tilt table that allows patients to perform a weight-bearing exercise, such as an inclined squat, in a gravity-reduced environment. Use of the modified tilt table may offer a more suitable therapeutic option when treating critically ill patients by providing a safe and controlled transition from bed rest to ambulation. Perhaps most importantly, the table appears to provide psychological benefits by empowering the patient to take more of an active role during the early stage of recovery. PMID- 12626930 TI - Alterations in functional movement after axillary burn scar contracture: a motion analysis study. AB - Children with axillary burns often develop scar contractures that restrict shoulder movement. Objective data on functional movement patterns after contracture formation is sparse. The purpose of this study was to determine how axillary contractures affect shoulder movement during activities of daily living (ADLs). This was a prospective study of children with axillary contractures scheduled for surgical release. Three-dimensional upper extremity kinematic analysis was used to assess shoulder, elbow, and trunk motion during two ADLs: high reach and hand to back pocket. Results were compared with a pool of 49 normal age-matched controls. Eleven children with axillary contractures were compared with controls. During high reach, significant decreases in shoulder flexion, shoulder internal rotation, arm pronation, and trunk extension occurred. Elbow flexion increased significantly. In the hand to back pocket task, shoulder extension and elbow flexion decreased and shoulder abduction increased. Axillary contractures result in quantifiable movement changes during ADLs. Aggressive rehabilitation is required to prevent contracture formation. Three-dimensional motion analysis is a unique tool for the quantification of functional limitations and provides an objective method to evaluate treatment efficacy in patients with axillary contractures. PMID- 12626931 TI - Psychosocial forum. PMID- 12626932 TI - Impact on the family: psychosocial adjustment of siblings of children who survive serious burns. AB - This study examined the psychosocial adjustment of 79 siblings of children suffering from burn injuries. Nonparametric statistics were used to compare psychosocial adjustment of the study group, as measured by the Child Behavior Checklist with an age-matched and gender-matched reference groups. Analyses found that the study group was better adjusted than the normative group on psychological dimensions; however, the study group fared worse than the normative group on overall competence, particularly social competence. Further analyses found significant differences in sibling adjustment as a function of the severity of the burn injury. The siblings of children with moderate burn injuries did significantly better on psychological adjustment than the normative group, and siblings of children with moderate and severe burn injuries did significantly poorer on social competence. Descriptive analysis of measures developed for the study for parent and sibling reports supported findings of the Child Behavior Checklist quantitative analysis and offered insight into reasons for findings. Results indicate that the burn injury to one child in a family significantly impacts the siblings of that child. The noninjured child may be strengthened in the process of adapting to the changes imposed on the family, but it is also possible that the sibling's growth in one dimension is at the cost of success in another dimension. PMID- 12626933 TI - Covering the uninsured. PMID- 12626934 TI - Smallpox vaccine: don't do it. PMID- 12626936 TI - House nursing caucus: the ANA applauds formation of a bipartisan congressional group. PMID- 12626935 TI - Breast cancer rates in black women. PMID- 12626937 TI - Reflections: Florida vacation. PMID- 12626938 TI - Safe patient handling and movement. PMID- 12626939 TI - The state of the science on urinary incontinence. PMID- 12626941 TI - Emergency: ED triage. PMID- 12626940 TI - Cultural considerations in end-of-life care: how ethnicity, age, and spirituality affect decisions when death is imminent. PMID- 12626944 TI - Professional respect: the CWPA and Magnet facilities work to improve nurse physician relationships. PMID- 12626946 TI - Giving the fingers a rest: alternative site testing eases blood glucose monitoring. PMID- 12626947 TI - Pollution: health care's unintended legacy. PMID- 12626948 TI - Pain management in patients with addictive disease. PMID- 12626949 TI - Screening for prostate cancer: recommendations and rationale. PMID- 12626951 TI - Negative behaviors in nursing. PMID- 12626952 TI - When the AIDS quilt came to Kirksville. PMID- 12626953 TI - Cadet Corps seeks Congressional recognition. PMID- 12626954 TI - Verbal abuse in the workplace. PMID- 12626955 TI - Research ethics and the intensive care unit: getting behind the wheel. PMID- 12626956 TI - Enrolling the uninsured in clinical trials: an ethical perspective. AB - Research participation by individuals who lack health insurance raises significant but seldom-discussed ethical issues for clinical investigators. Of particular concern are the possibility that their lack of options for getting care at the time of enrollment makes the uninsured susceptible to undue inducement to join trials and the possibility that their inability to access research products after trials leads to their exploitation. In tension with these concerns, however, is the right of the uninsured to fair consideration for research participation. We discuss these competing issues and make recommendations for how investigators can both protect uninsured research participants and provide the uninsured with fair access to research in their recruitment and enrollment strategies. PMID- 12626957 TI - Ethical framework for the use of sham procedures in clinical trials. AB - Ethical, professional, and financial considerations support the highest standards in demonstrating the safety and efficacy of new clinical interventions. Professional and regulatory mechanisms implemented to ensure that medical agents undergo a thorough process of systematic testing are lacking in the case of medical procedures. As a result, many procedures, including surgery, are rapidly introduced into the clinical setting without reliable evaluation of their efficacy. In many cases, a randomized, placebo-controlled procedure trial may be necessary to provide this data. We examine the major ethical objections to placebo-controlled procedure trials, including those involving sham surgery, and provide an ethical framework for assessing whether a placebo procedure is ethical to administer in the context of a clinical trial. A placebo-controlled trial of an invasive procedure can be ethically justified if: 1) there is a valuable, clinically relevant question to be answered by the research, 2) the placebo control is methodologically necessary to test the study hypothesis, 3) the risk of the placebo control itself has been minimized, 4) the risk of a placebo control does not exceed a threshold of acceptable research risk, 5) the risk of the placebo control is justified by valuable knowledge to be gained, and 6) the misleading involved in the administration of a placebo control is adequately disclosed and authorized during the informed consent process. PMID- 12626958 TI - Research in children. AB - The importance of medical research to the diagnosis and treatment of human diseases is well recognized. The use of human subjects, however, presents complex legal and ethical challenges for the scientific community and for society. The history of research performed on children reveals an especially vulnerable population needing special protection against violation of individual rights and exposure to undue risk. The development of guidelines and policies to protect children as research subjects is reviewed. Special focus is given to the present federal regulations that are intended to provide an ethical context for the performance of pediatric research, including the distinction between therapeutic and nontherapeutic studies. In part, these guidelines represent a return of the pendulum to a more moderate position, after an era of restrictive regulations in reaction to past abuses of children as research subjects. As a result, federal and professional initiatives are bringing renewed focus on the need for rigorous study of childhood development and disease within an appropriate ethical framework. PMID- 12626959 TI - Conflict of interest in human subjects research. PMID- 12626960 TI - Research ethics and law of healthcare system quality improvement: the conflict of cost containment and quality. AB - Quality improvement (QI) activities have been done as long as medicine has been practiced and are integral to safety and efficacy of patient care. The 20th century witnessed sophisticated advances of QI methods, with concurrent advances in research ethics. The suggestion that some interventional QI activities resemble research has led to a complex conundrum, with disparate forces driving for improvement and maintenance of patient care quality vs. the societal mandate to control healthcare costs. In the middle of these conflicting priorities are QI systems charged with effecting cost-effective and efficacious healthcare processes. Given the advances in research ethics, substantially grounded in the Nuremberg Code, and the increasing resemblance of interventional QI activities to research, the concern is raised of widespread and systematic Nuremberg Code violations occurring under the guise of QI in health care. Moreover, evidence is cited to suggest that if this is the case, then such activities may be subject to litigation or prosecutorial action. The ethical and legal foundations exist to support the systematic practice of informing patients of their participation in some types of QI procedures. PMID- 12626961 TI - Is the concept of informed consent applicable to clinical research involving critically ill patients? AB - Informed consent is a process through which patients or their surrogates authorize medical interventions or involvement in research. The concept of informed consent is supported by ethical principles and legal tradition in the United States. It grew in importance during the 20th century because of documented abuse of research subjects, which led to its codification in federal consent requirements for clinical research. For the most part, the concept and the codification are based on a model of competent patients who are capable of informed decision-making and can communicate their wishes. However, most critically ill patients are incompetent and cannot communicate easily. As a result, family members usually must give consent for them. Yet family members frequently are unavailable, and when they are available, they may not know the patient's wishes. Furthermore, family members may not be legally authorized to give consent for research involvement under state law. For these and other reasons, it has been argued that the concept of informed consent is not applicable to clinical research involving the critically ill and that such consent is not necessary in certain circumstances. Yet, for all its inadequacies, the concept of informed consent and the federal consent requirements should be retained because they promote respect for patients and their right of self determination and because investigator discretion is not adequate. Stronger research oversight may be as important as informed consent in protecting patient welfare. PMID- 12626962 TI - Educating researchers: ethics and the protection of human research participants. AB - To improve the protection of human participants in research, the federal government has mandated education and training in bioethics and issues relevant to human research. Despite the large time and financial commitments involved with such education, little is known about whether these efforts will actually improve the protection of human subjects. In this article, I review the history of ethics education in research leading up to the 2000 mandate. I then explore ethics education and its evaluation in the biological sciences and medicine and describe the previous successes and failures of these efforts. Many objectives can be the focus of educational interventions and evaluation. Some interventions in these fields had small, though statistically significant, effects on moral reasoning skills, knowledge, and confidence. Interventions are more likely to have lasting impact on moral reasoning if they were of moderate duration and involved small group discussion of dilemmas. Whether these measurable differences lead to changes in behavior or real-time application of moral reasoning skills remains to be determined. By having a clear understanding of the specific objectives, strengths, and limitations of an educational intervention, educators can design programs that may have an increased likelihood of improving protection of human research participants. PMID- 12626964 TI - Innovation to research: some transitional obstacles in critical care units. PMID- 12626963 TI - Critical care research on patients with advance directives or do-not-resuscitate status: ethical challenges for clinician-investigators. AB - Clinician-investigators face challenges in conducting research on critically ill patients when they have do-not-resuscitate orders, advance directives, or are in need of end-of-life care. Potential conflicts of interest for clinician investigators include either financial stakes or academic and reputational stakes. The dual roles for intensive care unit physician or nurse clinician investigators as healthcare professionals and scientists also present conflicts of interest, as does the dual purpose for the physical plant of the intensive care unit, which simultaneously serves as a site for patient care and a site for clinical research. Intensive care unit patients who become human research subjects also have dual roles that present conflict or confusion that can result in the therapeutic misconception. There are no scientifically or ethically sound reasons to exclude patients from participation in critical care research based on the presence of an advance directive or do-not-resuscitate order, as it would create a biased study sample that does not reflect the critically ill patient population, and it would treat a large group of potential research subjects differently from others without justification. There are four values in tension for critical care clinician-investigators in relation to patients/human research subjects: curative intent, palliative intent, research, and fiduciary obligations. A patient's decision to participate in research does not relieve clinician-investigators of their obligation to serve patient/human research subject's interests, even when doing so involves decisions to limit or withdraw life-sustaining interventions or withdraw the patient/human research subject from research. Critical care research involving patients with advance directives or do not-resuscitate status is both possible and desirable because it is just, respects patient autonomy, and results in study populations that better reflect the clinical population in all respects. PMID- 12626965 TI - Ethical considerations for research in critically ill patients. PMID- 12626967 TI - Mortality in Emergency Department Sepsis (MEDS) score: a prospectively derived and validated clinical prediction rule. AB - OBJECTIVES: Our objectives were a) to identify univariate correlates of death in emergency department patients at risk for infection; b) to perform multivariate analyses and identify independent predictors of death; and c) to develop and internally validate a prediction rule that may be used in the emergency department to risk stratify patients into different risk groups to predict their mortality rate. DESIGN: Prospective cohort study. SETTING: Emergency department of an urban university referral center. PATIENTS: Consecutive emergency department patients, aged 18 or older, who were at risk for infection, as indicated by the emergency department physician ordering a blood culture between February 1, 2000, and February 1, 2001. Of 3,301 eligible patient visits, 3,179 (96%) were enrolled. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The primary outcome was 28-day in-hospital mortality rate. There were 2,070 visits in the derivation set, with 110 deaths (5.3%), and 1,109 visits in the validation set, with 63 deaths (5.7%). Independent multivariate predictors of death were terminal illness (odds ratio, 6.1; 95% confidence interval, 3.6-10.2), tachypnea or hypoxia (2.7, 1.6-4.3), septic shock (2.7, 1.2-5.7), platelet count <150,000 (2.5, 1.5-4.3), band proportion >5% (2.3, 1.5-3.5), age >65 (2.2, 1.3-3.6), lower respiratory infection (1.9, 1.2-3.0), nursing home residence (1.9, 1.2-3.0), and altered mental status (1.6, 1.0-2.6). The clinical prediction rule stratified patients into mortality risk groups of very low, 0.9% (95% confidence interval, 0.2-1.5%); low, 2.0% (0.8-3.2%); moderate, 7.8% (5.6-10%); high, 20% (13-27%); and very high, 50% (36.1-64%) in the derivation set. Mortality rates for the corresponding risk groups in the validation set were 1.1%, 4.4%, 9.3%, 16%, and 39%, respectively. The receiver operating characteristic area for the rule was 0.82 in the derivation set and 0.78 in the validation set. CONCLUSIONS: In patients with suspected infection, this model identifies significant correlates of death and allows stratification of patients according to mortality risk. As new therapies become available for patients with sepsis syndromes, the ability to predict mortality risk may be helpful in triage and treatment decisions. PMID- 12626968 TI - Resolution of ventilator-associated pneumonia: prospective evaluation of the clinical pulmonary infection score as an early clinical predictor of outcome. AB - OBJECTIVES: To prospectively evaluate the performance of the Clinical Pulmonary Infection Score (CPIS) and its components to identify early in the hospital course of ventilator-associated pneumonia (VAP) which patients are responding to therapy. DESIGN: Prospective, multicenter, in a cohort of mechanically ventilated patients. SETTING: The intensive care unit of six hospitals located in the metropolitan area of Buenos Aires, Argentina. PATIENTS: Sixty-three patients, from a cohort of 472 mechanically ventilated patients hospitalized for >72 hrs, had clinical evidence of VAP and bacteriologic confirmation by bronchoalveolar lavage (BAL) or blood cultures. INTERVENTIONS: Bronchoscopy with BAL fluid culture and blood cultures after establishing a clinical diagnosis of VAP. All patients received antibiotics, 46 before bronchoscopy and 17 immediately after bronchoscopy. MEASUREMENTS AND RESULTS: CPIS was measured at 3 days before VAP (VAP-3); at the onset of VAP (VAP); and at 3 (VAP+3), 5 (VAP+5), and 7 (VAP+7) days after onset. CPIS rose from VAP-3 to VAP and then fell progressively in the population as a whole (p <.001), and the fall in CPIS was significant in 31 survivors, but not in 32 nonsurvivors. From the individual components of the CPIS, only the Pao /Fio ratio distinguished survivors from nonsurvivors, beginning at VAP+3. When CPIS was <6 at 3 or 5 days after VAP onset, mortality was lower than in the remaining patients (p =.018). These differences also related to the finding that those receiving adequate therapy had a slight fall in CPIS and a significant increase of Pao /Fio at VAP+3, whereas those getting inadequate therapy did not. CONCLUSIONS: Serial measurements of CPIS can define the clinical course of VAP resolution, identifying those with good outcome as early as day 3, and could possibly be of help to define strategies to shorten the duration of therapy. PMID- 12626969 TI - Leukotriene-mediated coronary vasoconstriction and loss of myocardial contractility evoked by low doses of Escherichia coli hemolysin in perfused rat hearts. AB - OBJECTIVE: hemolysin has been implicated as an important pathogenic factor in extraintestinal infections including sepsis. We investigated the effects of coronary administration of hemolysin on cardiac function in isolated rat hearts perfused at constant flow. DESIGN: Prospective, experimental study. SETTING: Research laboratory at a university hospital. SUBJECTS: Isolated hearts from male Wistar rats. INTERVENTIONS: Isolated hearts were perfused with purified hemolysin for 60 min. MEASUREMENTS AND MAIN RESULTS: Low concentrations of the toxin in the perfusate (0.1-0.2 hemolytic units/mL) caused a dose-dependent coronary vasoconstriction with a marked increase in coronary perfusion pressure, which was paralleled by a decrease in left ventricular developed pressure (and the maximum rate of left ventricular pressure increase). Moreover, 0.2 hemolytic units/mL hemolysin evoked ventricular fibrillation within 10 mins of toxin application. These events were accompanied by the liberation of leukotrienes (LTC4, LTD4, LTE4, and LTB4), thromboxane A2, prostaglandin I2, and the cell necrosis markers lactate dehydrogenase and creatine kinase into the recirculating perfusate. The lipoxygenase inhibitor MK-886 fully blocked the toxin-induced coronary vasoconstrictor response and the loss of myocardial contractility and reduced the release of lactate dehydrogenase and creatine kinase. In contrast to this, the cyclooxygenase inhibitor indomethacin was entirely ineffective. In addition, hemolysin elicited an increase in heart weight and left ventricular end-diastolic pressure, the latter again being suppressed by MK-886. CONCLUSIONS: Low doses of hemolysin cause strong coronary vasoconstriction, linked with loss of myocardial performance, release of cell injury enzymes, and electrical instability, with all events being largely attributable to toxin-elicited leukotriene generation in the coronary vasculature. Bacterial exotoxins such as hemolysin thus may be implicated in the cardiac abnormalities encountered in septic shock. PMID- 12626970 TI - Sex-based differences in serum cardiac troponin I, a specific marker for myocardial injury, after cardiac surgery. AB - BACKGROUND: Prevalence and causes of sex-based differences in morbidity and mortality secondary to cardiovascular disease remain controversial. Cardiac troponin I (cTnI) is a sensitive and specific marker for myocardial injury. Serial cTnI measurements have been used to identify perioperative myocardial cell injury. OBJECTIVE: To determine whether sex influences the extent of myocardial injury during cardiac surgery, we measured perioperative cTnI in male and female patients. DESIGN: A total of 17 male and 17 female patients were prospectively studied in an age- and case-matched manner. Arterial cTnI were obtained preinduction, 30 mins after the application of the aortic cross-clamp, at arrival to the intensive care unit, and on postoperative day 1. SETTING: Tertiary cardiac surgery center at a major teaching hospital. RESULTS: There was no difference between men and women in body mass index (kg/m2), duration of cardiopulmonary bypass, and aortic cross-clamp times. Preoperative cTnI measurements were similar in men (0.24 +/- 0.15 ng/mL) and women (0.25 +/- 0.13 ng/mL, mean +/- sem). The maximum serum cTnI occurred on postoperative day 1 in all patients, and it was 3 fold higher in men (18.5 +/- 5.7 ng/mL) compared with women (6.4 +/- 1.0 ng/mL). CONCLUSIONS: Men had markedly higher serum cTnI compared with women, although they were case matched with respect to age and cardiac risk factors. Our results may suggest there may be sex-related differences in the myocardial response to ischemia and reperfusion injury or intrinsic differences between the male and female myocardium. PMID- 12626971 TI - Effect of adverse drug reactions on length of stay in surgical intensive care units. AB - OBJECTIVE: To determine the frequency of adverse drug reactions in surgical intensive care units and evaluate their effect on the length of stay. DESIGN: Prospective cohort study. Between May 1997 and December 1999, while the patients were staying in the surgical intensive care unit, data were gathered regarding suspected adverse drug reactions and on different variables related to the length of stay. SETTING: Surgical intensive care units of our hospital. PATIENTS: A total of 401 patients hospitalized in the surgical intensive care unit. MAIN RESULTS: In 37 of the 401 patients seen (9.2%; 95% confidence interval, 6.6 12.5), 39 different adverse drug reactions were detected. The adverse drug reactions were most frequently caused by the following drugs: morphine hydrochloride (n = 13), meperidine hydrochloride (n = 9), and metamizole (n = 7). Five adverse drug reactions were severe, the suspected medication had to be discontinued in 14 cases, and new drugs were necessary to manage the adverse drug reaction in 28 cases. The crude estimation of the effect of adverse drug reactions performed on the length of stay with a bivariant regression model indicated that each adverse drug reaction was related to an increase of 3.39 days (95% confidence interval, 1.47-5.31) in the length of stay. This estimation was reduced to 2.31 days (95% confidence interval, 0.64-3.99) when considering other variables that might cause confusion for analysis, although it is still important. CONCLUSIONS: Adverse drug reactions are a significant clinical and economic problem in surgical intensive care units. PMID- 12626972 TI - Heat and moisture exchangers in mechanically ventilated intensive care unit patients: a plea for an independent assessment of their performance. AB - OBJECTIVE: To determine whether use of a hygroscopic and hydrophobic heat and moisture exchanger (HME) for 7 days without change affects its efficiency in long term, mechanically ventilated, chronic obstructive pulmonary disease (COPD) patients. DESIGN: Prospective, randomized, controlled clinical study comparing two combined HMEs. SETTING: Medical intensive care unit at a university teaching hospital. PATIENTS: Long-term, mechanically ventilated, COPD patients compared with non-COPD patients. INTERVENTIONS: In the first part of the study, COPD patients were studied with the Hygroster HME changed once a week. For the second part, the Hygroster was assessed in non-COPD patients and compared with the Hygrobac HME used in COPD and non-COPD patients for 1 wk without change. Devices could be changed if hygrometric measurements indicated insufficient humidity delivery. MEASUREMENTS AND MAIN RESULTS: Daily measurements were recorded for inspired gas temperature and relative and absolute humidity. Ventilatory variables, clinical indicators of efficient humidification, were also recorded. No tracheal tube occlusion occurred. However, contrary to the manufacturer advertisement, the Hygroster experienced surprisingly low values for absolute humidity in both COPD and non-COPD patients. Such events did not occur with the Hygrobac. Absolute humidity with the Hygroster was constantly and significantly lower during the 7-day study period than with the Hygrobac. Absolute humidity measured in COPD patients was identical to that measured in the rest of the study population with both HMEs. CONCLUSIONS: Manufacturer specifications and bedside measurements of absolute humidity differed considerably for the Hygroster, which in certain instances did not achieve efficient humidification in both COPD and non-COPD patients. This did not occur with the Hygrobac, which performed well throughout the 7-day period in both COPD and non-COPD patients. Our results speak for independent and evaluation of HMEs. PMID- 12626973 TI - Low exogenous lactate clearance as an early predictor of mortality in normolactatemic critically ill septic patients. AB - OBJECTIVE: To evaluate the prognostic value of lactate clearance and lactate production in severely ill septic patients with normal or mildly elevated blood lactate concentration. DESIGN Prospective, observational study. SETTING: Nineteen bed mixed medicosurgical intensive care unit. PATIENTS: Fifty-six patients with severe sepsis and blood lactate concentration <3 mmol/L. MEASUREMENTS AND MAIN RESULTS: Lactate metabolism was evaluated in all patients. Lactate clearance was measured by modeling the change in arterial blood lactate over time induced by an infusion of 1 mmol/kg sodium lactate for 15 mins. Lactate production was calculated as the product of lactate clearance times the blood lactate concentration before the infusion. Outcome was taken to be mortality at 28 days after the beginning of the septic episode. A logistic regression model taking into account different risk factors was constructed. Among the 56 patients, 17 (30.3%) died before the 28th day. Basal blood lactate concentration was not different between survivors and nonsurvivors, whereas lactate clearance and production were higher in survivors (0.86 +/- 0.32 vs. 0.58 +/- 0.18 L/hr/kg, p < .005, and 1.19 +/- 0.63 vs. 0.89 +/- 0.24 mmol/hr/kg, p = .055, respectively). An increase in blood lactate 45 mins after the end of the lactate infusion (Deltalact-T60) > or = 0.6 mmol/L was predictive of 28-day mortality with 53% sensitivity and 90% specificity. Multivariate analysis showed that only three factors were independently and significantly correlated with 28-day mortality: presence of more than two organ failures (odds ratio, 27; p = .04), age >70 yrs (odds ratio, 5.7; p = .032), and Deltalact-T60 > or =0.6 mmol/L (odds ratio, 14.2; p = .042). CONCLUSION: Low lactate clearance in severely ill septic patients with normal or mildly elevated blood lactate is predictive of poor outcome independently of other known risk factors such as age and number of organ failures. PMID- 12626974 TI - Safety of sedation with ketamine in severe head injury patients: comparison with sufentanil. AB - OBJECTIVE: The aim of the study was to compare the safety concerning cerebral hemodynamics of ketamine and sufentanil used for sedation of severe head injury patients, both drugs being used in combination with midazolam. DESIGN: Prospective, randomized, double-blind study. SETTING: Intensive care unit in a trauma center. PATIENTS: Twenty-five patients with severe head injury. INTERVENTIONS: Twelve patients received sedation with a continuous infusion of ketamine-midazolam and 13 with a continuous infusion of sufentanil-midazolam. All patients were mechanically ventilated with moderate hyperventilation. MEASUREMENTS AND MAIN RESULTS: Prognostic indicators (age, Glasgow Coma Scale scores, computed tomography diagnosis, and Injury Severity Scale score) were similar in the two groups at study entry. Measurements were carried out during the first 4 days of sedation. The average infusion rates during this time were 82 +/- 25 micro x kg x min ketamine and 1.64 +/- 0.5 microg x kg x min midazolam in the ketamine group and 0.008 +/- 0.002 microg x kg x min sufentanil and 1.63 +/- 0.37 microg x kg x min midazolam in the sufentanil group. No significant differences were observed between the two groups in the mean daily values of intracranial pressure and cerebral perfusion pressure. The numbers of intracranial pressure elevations were similar in both groups. The requirements of neuromuscular blocking agents, propofol, and thiopental were similar. Heart rate values were significantly higher in the ketamine group on therapy days 3 and 4 ( <.05). With regard to arterial pressure control, more fluids were given on the first therapy day and there was a trend toward greater use of vasopressors in the sufentanil group. Sedative costs were similar in the two groups. CONCLUSION: The results of this study suggest that ketamine in combination with midazolam is comparable with a combination of midazolam-sufentanil in maintaining intracranial pressure and cerebral perfusion pressure of severe head injury patients placed under controlled mechanical ventilation. PMID- 12626975 TI - Efficacy and safety of LY315920Na/S-5920, a selective inhibitor of 14-kDa group IIA secretory phospholipase A2, in patients with suspected sepsis and organ failure. AB - OBJECTIVE: Concentrations of group IIA secretory phospholipase A, an inflammatory response mediator, are increased in the plasma of patients with sepsis and septic shock, and the extent of elevation is correlated with mortality. LY315920Na/S 5920 is a selective inhibitor of group IIA secretory phospholipase A that has been shown to inhibit serum group IIA secretory phospholipase A enzyme activity in patients with severe sepsis. The primary objectives of this study were to determine whether there was a dose-response relationship between two doses of LY315920Na/S-5920 compared with placebo in the reduction of 28-day all-cause mortality in patients with severe sepsis and to determine whether LY315920Na/S 5920 had an acceptable safety profile.(2) (2) (2) DESIGN: Multicenter, double blind, placebo-controlled trial of two doses of LY315920Na/S-5920 in a parallel design. PATIENTS: A total of 586 patients with severe sepsis at 72 institutions in the United States. INTERVENTIONS Patients enrolled within 72 hrs from onset of first sepsis-induced organ failure were randomized (1:1:1) to low-dose LY315920Na/S-5920 (target plasma concentration of 200 ng/mL, n = 196), high-dose LY315920Na/S-5920 (800 ng/mL, n = 194), or placebo (n = 196). Study medication was administered as a constant-rate intravenous infusion for 168 hrs. MEASUREMENTS AND MAIN RESULTS: The study was stopped prematurely because it was unlikely that a statistically significant difference in mortality between LY315920Na/S-5920 and placebo would be found. There was no effect of LY315920Na/S 5920 on the primary end point of 28-day all-cause mortality across the entire study population. The 28-day all-cause mortality was distributed as follows: placebo group, 33.2% (65/196 patients); low-dose LY315920Na/S-5920, 37.2% (73/196); and high-dose LY315920Na/S-5920, 36.1% (70/194); p = .525. However, in a prospectively planned analysis, there was a favorable overall dose-response effect on 28-day all-cause mortality in patients administered LY315920Na/S-5920 within 18 hrs of onset of the first sepsis-induced organ failure. Among these patients, 28-day all-cause mortality was distributed as follows: placebo group, 43.5% (20/46 patients); low-dose LY315920Na/S-5920, 31.4% (16/51); and high-dose LY315920Na/S-5920, 20.8% (10/48); p = .018. CONCLUSIONS: Administration of LY315920Na/S-5920 had an acceptable safety profile in patients with severe sepsis. There was no overall survival benefit associated with the use of LY315920Na/S-5920 in this study. However, prospectively planned secondary analyses suggested that treatment with LY315920Na/S-5920 was associated with an improvement in survival in patients treated within 18 hrs of the first sepsis induced organ failure. PMID- 12626976 TI - Bacterial colonization of the respiratory tract following tracheal intubation effect of gravity: an experimental study. AB - OBJECTIVE: To explore the role of the horizontal orientation of endotracheal tube and neck on bacterial colonization of the respiratory tract in anesthetized sheep on mechanical ventilation, without use of antibiotics. DESIGN: Prospective animal study. SETTING: National Institutes of Health research laboratory. SUBJECTS: Anesthetized, paralyzed, and ventilated sheep. INTERVENTIONS: Sheep were randomized into five groups and managed as follows: Group IS contained sheep that were not intubated and were immediately killed. Group HU4 contained six sheep that were mechanically ventilated for 4 hrs, with head and endotracheal tube elevated 30 degrees from horizontal. Group HU72 contained seven sheep that were prone, mechanically ventilated for 72 hrs, and managed the same as group HU4. Groups G and Gf each contained seven sheep that were prone on a lateral body rotation device, mechanically ventilated for 72 hrs, with neck and endotracheal tube horizontal. Group Gf received nasogastric enteral feeding. MEASUREMENTS AND MAIN RESULTS: At the end of the study, sheep were examined postmortem, and a total of 11 tissue samples were taken from the trachea, the five lobar bronchi, and the five lobar parenchyma, for qualitative and quantitative culture. Group HU72 had significant decrease in Pao2/Fio2 and heavy bacterial colonization in all sheep. Groups G and Gf retained excellent lung function; lung bacterial colonization was no different from the IS group. CONCLUSIONS: The horizontal orientation of the endotracheal tube and neck, through lateral body rotation, showed no altered airway colonization and maintained excellent gas exchange and lung function in our animal model. PMID- 12626977 TI - Effects of sustained inflation and postinflation positive end-expiratory pressure in acute respiratory distress syndrome: focusing on pulmonary and extrapulmonary forms. AB - OBJECTIVE: To investigate whether the response to sustained inflation and postinflation positive end-expiratory pressure varies between acute respiratory distress syndrome with pulmonary (ARDS(exp)) and extrapulmonary origin (ARDS(exp)). DESIGN: Prospective clinical study. SETTING: Multidisciplinary intensive care unit in a university hospital. PATIENTS: A total of 11 patients with ARDS and 13 patients with ARDS. INTERVENTIONS: A 7 ml/kg tidal volume, 12-15 breaths/min respiratory rate, and an inspiratory/expiratory ratio of 1:2 was used during baseline ventilation. Positive end-expiratory pressure levels were set according to the decision of the primary physician. Sustained inflation was performed by 45 cm H2O continuous positive airway pressure for 30 secs. Postinflation positive end-expiratory pressure was titrated decrementally, starting from a level of 20 cm H2O to keep the peripheral oxygen saturation between 92% and 95%. Fio2 was decreased, and baseline tidal volume, respiratory rate, inspiratory/expiratory ratio were maintained unchanged throughout the study period. MEASUREMENTS AND MAIN RESULTS: Blood gas, airway pressure, and hemodynamic measurements were performed at the following time points: at baseline and at 15 mins, 1 hr, 4 hrs, and 6 hrs after sustained inflation. After sustained inflation, the Pao2/Fio2 ratio improved in all of the patients both in ARDS(p) and ARDS(exp). However, the Pao2/Fio2 ratio increased to >200 in four ARDS(p) patients (36%) and in seven ARDS(p) patients (54%). In two of those ARDS patients, the Pao2/Fio2 ratio was found to be <200, whereas none of the ARDS(p) patients revealed Pao2/Fio2 ratios of <200 at the 6-hr measurement. Postinflation positive end-expiratory pressure levels were set at 16.7 +/- 2.3 cm H O in ARDS(p) and 15.6 +/- 2.5 cm H2O in ARDS. The change in Pao /Fio ratios was found statistically significant in patients with ARDS(p) (p =.0001) and with ARDS(p) (p =.008). Respiratory system compliance increased in ARDS patients (p =.02), whereas the change in ARDS was not statistically significant. CONCLUSIONS: Sustained inflation followed by high levels of postinflation positive end expiratory pressure provided an increase in respiratory system compliance in ARDS; however, arterial oxygenation improved in both ARDS forms. PMID- 12626978 TI - Right atrial pressure as a measure of ventricular constraint arising from positive end-expiratory pressure during mechanical ventilation of the neonatal lamb. AB - OBJECTIVE: By constraining the heart, the chest wall, lungs, and pericardium limit diastolic filling and thus have a major role in determining cardiac output. Although intermittent positive pressure ventilation and the application of positive end-expiratory pressure amplifies this constraint, no clinical method exists to assess the impact that positive end-expiratory pressure has on ventricular constraint in the newborn. In this study, we tested the hypothesis that a change in right atrial pressure (Pra) reflects the change in ventricular constraint associated with a change in airway pressure. DESIGN: Experimental, comparative animal study. SETTING: Ritchie Centre for Baby Health Research cardiovascular laboratory. SUBJECTS Neonatal (4-wk-old, n = 6) and newborn (3-day old, n = 6) Merino/Border-Leicester cross lambs. INTERVENTIONS: Lambs were anesthetized (alpha-chloralose and ketamine), ventilated, and instrumented to record Pra, thoracic inferior vena caval pressure (Pivc, saline-filled catheters), and pericardial pressure (Pper, liquid-containing balloon). MEASUREMENTS AND MAIN RESULTS: Changes (Delta) in Pra, Pivc, and Pper were assessed while airway pressure was rapidly reduced from four set levels of continuous positive airway pressure (2.5, 5, 7.5, and 15 cm H2O) to atmospheric pressure. A strong linear relationship was observed between DeltaPra and DeltaPper (DeltaPra = 0.90 DeltaPper - 0.02, r =.98), and between DeltaPivc and DeltaPper (DeltaPivc = 0.86 DeltaPper - 0.02, r =.98) in both the 4-wk-old lambs and the 3-day-old lambs. CONCLUSIONS: Our experiments demonstrate that, in the newborn and neonatal lamb, DeltaPra provides an accurate measure of the change in ventricular constraint that accompanies a change in airway pressure, and thus may provide a means of quantifying the magnitude of ventricular constraint imposed by positive end-expiratory pressure and mechanical ventilation during neonatal intensive care. PMID- 12626979 TI - Can yeast isolation in peritoneal fluid be predicted in intensive care unit patients with peritonitis? AB - OBJECTIVE: To generate and validate a predictive score of yeast isolation based on independent risk factors of yeast isolation in intensive care unit patients with peritonitis. DESIGN: Retrospective cohort study to determine independent risk factors of yeast isolation, generation of the score, and validation in a prospective cohort of patients with peritonitis. SETTING: Tertiary-care, university-affiliated hospital. PATIENTS: Two hundred twenty-one patients with peritonitis hospitalized in a surgical intensive care unit between 1994 and 1999 for the retrospective cohort and 57 patients in the prospective cohort (2000). MEASUREMENTS AND MAIN RESULTS: Four independent risk factors of yeast isolation in peritoneal fluid (similar odds ratio) were found in the retrospective cohort: female gender, upper gastrointestinal tract origin of peritonitis, intraoperative cardiovascular failure, and previous antimicrobial therapy at least 48 hrs before the onset of peritonitis. A score based on the number of risk factors was constructed (grade A = zero or one risk factor, grade B = at least two risk factors, grade C = at least three risk factors, and grade D = four risk factors), and validated in the prospective cohort. For a grade C score, sensitivity was 84%, specificity was 50%, positive and negative predictive values were 67% and 72%, respectively, and overall accuracy was 71%. CONCLUSIONS: Four independent risk factors of yeast isolation in the peritoneal fluid were identified in critically ill surgical patients with peritonitis. The presence of at least three of these factors (grade C score) was associated with a high rate of yeast detection. This approach could be helpful to initiate early antifungal therapy in this patient population. PMID- 12626980 TI - Experience with percutaneous venoarterial cardiopulmonary bypass for emergency circulatory support. AB - OBJECTIVE: Mechanical circulatory support can maintain vital organ perfusion in patients with cardiac failure unresponsive to standard pharmacologic treatment. The purpose of the current study was to report complication and survival rates in patients supported with emergency percutaneous venoarterial cardiopulmonary bypass because of prolonged cardiogenic shock or cardiopulmonary arrest. DESIGN: Retrospective clinical study. SUBJECTS: A total of 46 patients supported with venoarterial cardiopulmonary bypass, 25 because of cardiogenic shock unresponsive to pharmacologic therapy and 21 because of cardiopulmonary arrest unresponsive to standard advanced cardiac life support. RESULTS: In 41 of the 46 patients (89%), stable extracorporeal circulation was established; in five patients (11%), femoral cannulation was accomplished only after a surgical cutdown. A total of 28 patients were weaned from cardiopulmonary bypass (19 of 25 patients with cardiogenic shock vs. 9 of 21 patients with cardiopulmonary arrest, p =.03), and 13 patients had long-term survival (10 of 25 patients with cardiogenic shock vs. 3 of 21 patients with cardiopulmonary arrest, p =.1). Complications directly related to the use of cardiopulmonary bypass were found in 18 patients (39%), major complications related to femoral cannulation being the most common single cause for bypass-associated morbidity (eight patients, 17%). CONCLUSIONS: Long term survival rates after emergency percutaneous cardiopulmonary bypass are encouraging in patients with an underlying cardiocirculatory disease amenable to immediate corrective intervention (angioplasty, surgery, transplantation). PMID- 12626981 TI - Low stretch ventilation strategy in acute respiratory distress syndrome: eight years of clinical experience in a single center. AB - CONTEXT: In recent years, protective ventilation with airway pressure limitation has constituted a major advance in acute respiratory distress syndrome treatment and has led to a substantial improvement in prognosis. With this therapeutic rationale, one may even question nowadays whether the severity of respiratory failure still influences mortality. OBJECTIVE: To determine whether the severity of respiratory failure, scored according to the usual criteria, still influences mortality in acute respiratory distress syndrome patients when a low stretch ventilation was used and to assess the impact on mortality of other nonpulmonary organ dysfunction, particularly circulatory failure. DESIGN AND SETTING: A retrospective study conducted in the medical intensive care unit of a French university hospital from October 1993 to December 2001. PATIENTS A total of 150 acute respiratory distress syndrome patients who were administered uniform protective ventilation with a limited plateau pressure (<30 cm H2O), a low positive end-expiratory pressure (<10 cm H2O), and the same strategy concerning hemodynamic support and dialysis when required. MAIN OUTCOME AND MEASURES: Mean age, general severity index (Simplified Acute Physiologic Score II), number of associated organ failures (Logistic Organ Dysfunction Score), respiratory severity indices (Pao2/Fio2, Lung Injury Severity Score), and severity of initial circulatory failure (circulatory failure present at admission or that developed during the first 48 hrs) were compared, according to recovery or death, and evaluated by a logistic regression model, which allows simultaneous control of multiple factors. RESULTS: The average mortality rate for the whole group was 38%, with 93 patients recovering after an average duration of mechanical ventilation of 18 +/- 13 days. The major factor significantly and independently associated with probability of dying was the severity of circulatory failure (p =.0001, odds ratio = 10.17). Patients free from initial circulatory failure (39 patients) had a 95% recovery rate. CONCLUSION: With our low stretch strategy, the severity of circulatory failure was the main determinant of acute respiratory distress syndrome prognosis. Patients with isolated respiratory failure during the first 48 hrs of respiratory support have an excellent chance of recovery when treated with protective ventilation associated with a low positive end-expiratory pressure. PMID- 12626982 TI - Platelet-activating factor acetylhydrolase is increased in lung lavage fluid from patients with acute respiratory distress syndrome. AB - OBJECTIVE: Platelet-activating factor (PAF) is a proinflammatory phospholipid that may contribute to inflammation in the acute respiratory distress syndrome (ARDS). PAF acetylhydrolase (PAF-AH) degrades PAF and regulates its biological activity. We characterized PAF-AH in bronchoalveolar lavage fluid from ARDS patients (n = 33, 22 survivors), patients at risk for ARDS (n = 6), and healthy controls (n = 6). DESIGN: Bronchoalveolar lavage was performed during acute (<96 hrs from onset), plateau (6 to 12 days), and late (> or = 14 days) phases of ARDS. PATIENTS: Intubated patients with ARDS or a risk factor for ARDS. MEASUREMENTS AND MAIN RESULTS: In ARDS, total bronchoalveolar lavage PAF-AH activity was markedly increased in the acute phase (87 +/- 89 mU/mL, n = 33) and then decreased in the plateau (23 +/- 14 mU/mL, n = 10) and late phases (19 +/- 14 mU/mL, n = 7) (p = .003). Total bronchoalveolar lavage PAF-AH activity during the acute phase of ARDS was also increased as compared with patients at risk for ARDS (16 +/- 13 mU/mL, n = 6) and healthy controls (3 +/- 3 mU/mL, n = 6) (p < .001). In contrast, plasma PAF-AH activities were the same in controls (3215 +/- 858 mU/mL, n = 6), in patients at risk for ARDS (3606 +/- 1607 mU/mL, n = 6), and during the acute phase of ARDS (3098 +/- 2395 mU/mL, n = 33) (p = .18). PAF-AH mRNA was present in alveolar macrophages in the acute phase of ARDS (five of six) and in at-risk patients (two of three) but not in healthy controls. CONCLUSIONS: PAF-AH activity is increased in bronchoalveolar lavage fluid from patients with ARDS. Likely sources include leakage of plasma PAF-AH into alveoli or release of PAF-AH from injured cells; however, the presence of PAF-AH mRNA in alveolar macrophages suggests that PAF-AH may be actively synthesized in the lungs of patients with ARDS. PAF-AH activity in the lungs of ARDS patients may regulate inflammation caused by PAF and related oxidized phospholipids generated in the inflammatory response. PMID- 12626984 TI - Inadequate salivary flow and poor oral mucosal status in intubated intensive care unit patients. AB - OBJECTIVE: To investigate salivary flow and frequency of oral mucositis in intensive care unit patients compared with patients admitted because of elective coronary artery bypass graft (CABG) surgery. In addition, the pattern of oropharyngeal colonization was investigated in these patients. DESIGN: Prospective study. SETTING: Mixed intensive care unit and cardiosurgical ward. PATIENTS: In this study, 24 ventilated intensive care unit patients and 20 CABG patients were included. MEASUREMENTS AND MAIN RESULTS: Two dental hygienists examined intensive care unit patients for the presence of periodontal disease and mucositis at admission and subsequently every week during their stay in the intensive care unit. At the same time, stimulated salivary flow and salivary total immunoglobulin A output were measured. Oropharyngeal cultures were obtained as well. CABG patients were examined the day before the operation, 1 day, 1 wk, and 2 wks after the operation. The following results were obtained: a) temporarily reduced postoperative stimulated salivary flow and total salivary immunoglobulin A output in CABG patients and nearly absent stimulated salivary flow in intensive care unit patients; b) oropharyngeal colonization with potentially pathogenic microorganisms in intensive care unit and not in CABG patients; and c) the increase in mucositis index in intensive care unit patients paralleled the increase in potentially pathogenic microorganism oropharyngeal colonization, especially and. CONCLUSIONS: Absence of adequate salivary flow in intubated intensive care unit patients causes severe xerostomia, which may contribute to the development of mucositis and oropharyngeal colonization with Gram-negative bacteria. PMID- 12626983 TI - Enteral naloxone reduces gastric tube reflux and frequency of pneumonia in critical care patients during opioid analgesia. AB - OBJECTIVE: Opioid analgesia impairs gastrointestinal motility. Enteral administration of naloxone theoretically allows selective blocking of intestinal opioid receptors caused by extensive presystemic metabolism. Therefore, we studied the effect of enteral naloxone on the amount of gastric tube reflux, the frequency of pneumonia, and the time until first defecation in mechanically ventilated patients with fentanyl analgesia. DESIGN: Prospective, randomized, double-blinded study. SETTING: University hospital intensive care unit. PATIENTS: Eighty-four mechanically ventilated, fentanyl-treated patients without gastrointestinal surgery or diseases. INTERVENTIONS: Patients were assigned to receive 8 mg naloxone or placebo four times daily via a gastric tube during fentanyl administration. MEASUREMENTS AND MAIN RESULTS: Thirty-eight patients received naloxone and 43 placebo; three patients were excluded because of protocol violation. Median gastric tube reflux volume (54 vs. 129 mL, p =.03) and frequency of pneumonia (34% vs. 56%, p =.04) were significantly lower in the naloxone group. In both groups, time until first defecation, ventilation time, and length of intensive care unit stay did not differ. There was no difference in fentanyl requirements between the naloxone and the placebo group (7 vs. 6.5 microg/kg/hr, p =.15). CONCLUSIONS: Our results provide evidence that the administration of enteral opioid antagonists in ventilated patients with opioid analgesia might be a simple-and possibly preventive-treatment of increased gastric tube reflux and reduces frequency of pneumonia. PMID- 12626985 TI - Free radical and drug oxidation products in an intensive care unit sedative: propofol with sulfite. AB - OBJECTIVES: Some propofol emulsion formulations contain EDTA or sodium metabisulfite to inhibit microbe growth on extrinsic contamination. EDTA is not known to react with propofol formulation components; however, sulfite has been shown to support some oxidation processes and may react with propofol. This study compared the oxidation of propofol and the formation of free radicals by electron paramagnetic resonance analysis in EDTA and sulfite propofol emulsions during a simulated intensive care unit 12-hr intravenous infusion. DESIGN: Controlled laboratory study. SETTING: University laboratory. MEASUREMENTS AND MAIN RESULTS: Propofol emulsions (3.5 mL) were dripped from spiked 50-mL vials at each hour for 12 hrs. Two propofol oxidation products, identified as propofol dimer and propofol dimer quinone, were detected in sulfite and EDTA propofol emulsions; however, sulfite propofol emulsion contained higher quantities of both compounds. After initiation of the simulated infusion, the quantities of propofol dimer and propofol dimer quinone increased in the sulfite propofol emulsion, but the lower levels in the EDTA propofol emulsion remained constant. Sulfite propofol emulsion began to visibly yellow at about 6-7 hrs. The EDTA propofol emulsion remained white at all times. The absorbance spectra of the propofol dimer and propofol dimer quinone extracted from sulfite propofol emulsion showed that propofol dimer did not absorb in the visible spectrum, but the propofol dimer quinone had an absorbance peak at 421 nm, causing it to appear yellow. Electron paramagnetic resonance analysis of the propofol emulsion containing metabisulfite revealed that the sulfite propofol emulsion yielded a strong free radical signal consistent with the formation of the sulfite anion radical (SO3*-). The EDTA propofol emulsion yielded no free radical signal above background. CONCLUSION: Sulfite from the metabisulfite additive in propofol emulsion creates an oxidative environment when these emulsions are exposed to air during a simulated intravenous infusion. This oxidation results in propofol dimerization and emulsion yellowing, the latter of which is caused by the formation of propofol dimer quinone. These processes can be attributed to the rapid formation of the reactive sulfite free radical. PMID- 12626986 TI - Factors influencing the accuracy of oscillometric blood pressure measurement in critically ill patients. AB - OBJECTIVE: Comparison of oscillometric blood pressure measurement with two different devices (M3000A using a new algorithm and M1008A using an established algorithm, both Hewlett Packard) and evaluation of current recommendations concerning the relation between cuff size and upper arm circumference in critically ill patients. DESIGN: Prospective data collection. SETTING: Emergency department in a 2000-bed inner-city hospital. PATIENTS: A total of 30 patients categorized into three groups according to their upper arm circumference (I, 18 25 cm; II, 25.1-33 cm; III, 33.1-47.5 cm) were enrolled in the study protocol. INTERVENTIONS In each patient, two noninvasive blood pressure devices with three different cuff sizes were used to perform oscillometric blood pressure measurement. Invasive mean arterial blood pressure measurement was done by cannulation of the radial artery. MEASUREMENT AND MAIN RESULTS: Overall, 1,011 pairs of simultaneous oscillometric and invasive blood pressure measurements were collected in 30 patients (group I, n = 10; group II, n = 10; group III, n = 10). The overall discrepancy between both methods with the M3000A was -2.4 +/- 11.8 mm Hg (p <.0001) and, with the M1008A, -5.3 +/- 11.6 mm Hg (p <.0001) if the recommended cuff size according to the upper arm circumference was used (352 measurements). If smaller cuff sizes than recommended were used (308 measurements performed in group II and III), the overall discrepancy between both methods with the M3000A was 1.3 +/- 13.4 mm Hg (p <.024) and, with the M1008A, -2.3 +/- 11.5 mm Hg (p <.0001). CONCLUSION: The new algorithm reduced the overall bias of the oscillometric method but still showed a significant discrepancy between both methods of blood pressure measurement, primarily due to the mismatch between upper arm circumference and cuff size. The improvement of the algorithm alone could not result in a sufficient improvement of oscillometric blood pressure measurement. A reevaluation of the recommendations concerning the relation between upper arm circumference and cuff size are urgently required if oscillometric blood pressure measurement should become a reasonable alternative to intra-arterial blood pressure measurement in critically ill patients. PMID- 12626987 TI - Comparison of different airway management strategies to ventilate apneic, nonpreoxygenated patients. AB - OBJECTIVE: Endotracheal intubation is the gold standard for providing emergency ventilation, but acquiring and maintaining intubation skills may be difficult. Recent reports indicate that even in urban emergency medical services with a high call volume, esophageal intubations were observed, requiring either perfect intubation skills or development of alternatives for emergency ventilation. DESIGN: Simulated emergency ventilation in apneic patients employing four different airway devices that used small tidal volumes. SETTING: University hospital operating room. SUBJECTS: Forty-eight ASA I/II patients who signed written informed consent before being enrolled into the study. INTERVENTIONS: In healthy adult patients without underlying respiratory or cardiac disease who were breathing room air before undergoing routine induction of surgery, 12 experienced professional paramedics inserted either a laryngeal mask airway (n = 12), Combitube (n = 12), or cuffed oropharyngeal airway (n = 12) or placed a face mask (n = 12) before providing ventilation with a pediatric (maximum volume, 700 mL) self-inflating bag with 100% oxygen for 3 mins. MEASUREMENTS AND MAIN RESULTS: In three of 12 cuffed oropharyngeal airway patients, two of 12 laryngeal mask airway patients, and one of 12 Combitube patients, oxygen saturation fell below 90% during airway device insertion, and the experiment was terminated; no oxygenation failures occurred with the bag-valve-mask. Oxygen saturation decreased significantly (p <.05) during insertion of the Combitube and laryngeal mask but not with the bag-valve-mask and cuffed oropharyngeal airway; however, oxygen saturation increased after 1 min of ventilation with 100% oxygen. No differences in tidal lung volumes were observed between airway devices. CONCLUSIONS: Paramedics were able to employ the laryngeal mask airway, Combitube, and cuffed oropharyngeal airway in apneic patients with normal lung compliance and airways. In this population, bag-valve-mask ventilation was the most simple and successful strategy. Small tidal volumes applied with a pediatric self-inflating bag and 100% oxygen resulted in adequate oxygenation and ventilation. PMID- 12626988 TI - Plasma concentrations of von Willebrand factor and intracellular adhesion molecule-1 for prediction of outcome after successful cardiopulmonary resuscitation. AB - OBJECTIVE: Ischemia/reoxygenation following cardiopulmonary resuscitation might cause endothelial injury/activation that could contribute to an adverse outcome after cardiopulmonary resuscitation. We studied plasma concentrations of von Willebrand factor (vWF) antigen and soluble intracellular adhesion molecule (sICAM)-1 as markers of a generalized endothelial injury/activation in relation to outcome after cardiopulmonary resuscitation. DESIGN: Retrospective study on stored plasma samples. SETTING: Intensive care unit at a university hospital. PATIENTS: Thirty-five patients who survived >24 hrs after in- or out-of-hospital cardiopulmonary resuscitation and 15 noncritically ill control patients.I NTERVENTIONS: Blood sampling. MEASUREMENTS AND MAIN RESULTS: Plasma concentrations of vWF antigen and sICAM-1 on day 2 after cardiopulmonary resuscitation were higher in patients than in controls (p < .001 and p = .001, respectively). In-hospital cardiopulmonary resuscitation, cardiopulmonary resuscitation duration > or = 15 mins, severe cardiovascular failure, and renal dysfunction/failure at the time of blood sampling were associated with significant elevations in vWF antigen and sICAM-1 concentrations. Patients with an unfavorable outcome after cardiopulmonary resuscitation (cerebral performance category > or = 3) exhibited higher vWF antigen and sICAM-1 concentrations than patients with good outcome (cerebral performance category 1-2; p < .001 and p = .097, respectively). Renal dysfunction/failure, severe cardiovascular failure, systemic inflammatory response syndrome, and cardiopulmonary resuscitation duration > or = 15 mins were also associated with higher adverse outcome rates. Combination of these four variables into a cardiac arrest risk score (levels 0-4) showed adverse outcome rates of 100, 56, and 0% in patients with arrest scores of 4, 2-3, and 0-1, respectively. A vWF antigen concentration >166% was an independent predictor of outcome after cardiopulmonary resuscitation (p = .002) and was associated with increased adverse outcome rates in patients with cardiac arrest risk scores of 2-3. Furthermore, both vWF antigen concentrations >166% and sICAM-1 concentrations >500 ng/mL had 100% specificity for an adverse outcome in patients after out-of-hospital cardiopulmonary resuscitation but were less predictive in patients after in-hospital cardiopulmonary resuscitation. CONCLUSIONS: vWF antigen and sICAM-1 might be useful adjunctive variables for early determination of outcome in patients after successful out-of-hospital cardiopulmonary resuscitation. PMID- 12626989 TI - Bronchoalveolar interleukin-1 beta: a marker of bacterial burden in mechanically ventilated patients with community-acquired pneumonia. AB - OBJECTIVE: To assess the relationship between concentrations of bronchoalveolar cytokines and bacterial burden (quantitative bacterial count) in intubated patients with a presumptive diagnosis of community-acquired pneumonia. DESIGN: A cross-sectional and clinical investigation. SETTING Medical/surgical and respiratory intensive care unit of a tertiary 1,200-bed medical center. PATIENTS: According to the time course of community-acquired pneumonia at the time of study with bronchoalveolar lavage, 69 mechanically ventilated patients were divided into three subgroups: primary (n = 11), referral (n = 23), and treated (n = 35) community-acquired pneumonia. INTERVENTIONS: Bronchoalveolar lavage was performed in the most abnormal area on chest radiograph by fiberoptic bronchoscope. Bronchoalveolar lavage fluid was processed for quantitative bacterial culture. The concentrations of bronchoalveolar lavage cytokines (tumor necrosis factor alpha, interleukin-1 beta, interleukin-6, interleukin-8, and interleukin-10) also were measured. MEASUREMENTS AND MAIN RESULTS: Thirty-two patients had a positive bacterial culture (bronchoalveolar lavage > or = 10 colony-forming units/mL)., and made up 76% of pathogens recovered at high concentrations. The concentrations of bronchoalveolar lavage interleukin-1 beta were 199.1 +/- 32.1 and 54.9 +/- 13.0 pg/mL (mean +/- se) in the patients with positive and negative bacterial culture, respectively (p < .001). Bronchoalveolar lavage interleukin- 1 beta was significantly higher in the patients with a high bacterial burden (p < .001), with mixed bacterial infection (p < .001), and with pneumonia (p < .001), compared with values in patients without these features. The relationship between bacterial load and concentrations of bronchoalveolar lavage interleukin-1 beta was very strong in the patients with primary and referral community-acquired pneumonia but was borderline in treated community-acquired pneumonia. CONCLUSIONS: The common pathogens were similar to the core pathogens of hospital acquired pneumonia, probably due to antibiotic effects, delayed sampling, and superimposed nosocomial infection. Since the concentration of bronchoalveolar lavage interleukin-1 beta was correlated with bacterial burden in the alveoli, it may be a marker for progressive and ongoing inflammation in patients who have not responded to pneumonia therapy and who have persistence of bacteria in the lung. PMID- 12626990 TI - Sublingual capnometry versus traditional markers of tissue oxygenation in critically ill patients. AB - OBJECTIVE: The purpose of this study was to determine the prognostic value of sublingual PCO2 (P(SL)CO2), lactate concentration, and mixed venous oxygen saturation (S(MV)O2) in hemodynamically unstable intensive care patients and, additionally, to compare the temporal changes of these variables in response to treatment. SETTING: Medical/surgical intensive care unit. SUBJECTS: Fifty-four patients, mean age 58 +/- 8 yrs. INTERVENTIONS: Oxyhemodynamic variables, arterial lactate concentration, and P(SL)CO2 were recorded in unselected sequential intensive care patients undergoing pulmonary artery catheterization. A data set was obtained immediately after insertion of the pulmonary artery catheter and repeated 4 and 8 hrs later. MEASUREMENTS AND MAIN RESULTS: Twenty one patients had severe sepsis or septic shock. Twenty-seven (50%) patients died. The initial P(SL)CO2_PaCO2 gradient (P(SL)CO2-diff) and the initial P(SL)CO2 were highly predictive of outcome (p =.0004 and p =.004, respectively); however, there was no difference in the arterial lactate concentration and S(MV)O2 between the survivors and nonsurvivors. The P(SL)CO2-diff had the best receiver operator characteristic characteristics (area under the curve, 0.75), with a P(SL)CO2-diff >25 mm Hg being the best discriminator of outcome. With treatment, the P(SL)CO2 diff decreased in both survivors and nonsurvivors; however, the lactate and S(MV)O2 remained relatively unchanged during the study period. CONCLUSIONS: The baseline P(SL)CO2-diff and P(SL)CO2 were better predictors of outcome than traditional markers of tissue hypoxia and were more responsive to therapeutic interventions. The P(SL)CO2-diff and/or P(SL)CO2 may prove to be a useful marker for goal-directed therapy and for assessing the response to clinical interventions aimed at improving tissue oxygenation. PMID- 12626991 TI - Long-term mechanical ventilation with hygroscopic heat and moisture exchangers used for 48 hours: a prospective clinical, hygrometric, and bacteriologic study. AB - OBJECTIVE: To determine whether use of a hygroscopic heat and moisture exchanger (HME) for 48 hrs without change affects its efficiency and the level of bacterial colonization in long-term mechanically ventilated medical intensive care unit patients. DESIGN: Prospective, randomized clinical study evaluating two hygroscopic HMEs. SETTING: Medical intensive care unit at a university teaching hospital. PATIENTS: Long-term mechanically ventilated medical intensive care unit patients, including chronic obstructive pulmonary disease patients. INTERVENTIONS: Patients were randomly allocated to one of the two HMEs studied (Hygrolife and EdithFlex) and changed every 48 hrs. Devices in both groups could be changed if hygrometric measurements indicated insufficient humidity delivery. MEASUREMENTS AND MAIN RESULTS: Daily measurements of inspired gas temperature and relative and absolute humidity. In addition, cultures of tracheal aspirations and both patient and ventilator sides of the device were performed after 48 hrs of use. Ventilatory variables and clinical indicators of efficient humidification were also recorded. Prolonged use of both HMEs was safe and efficient (no tracheal tube occlusion occurred). Mean duration of mechanical ventilation was 20 days. Both clinical indicators and hygrometric measurements showed that both devices performed well during 48 hrs. Absolute humidity with EdithFlex was significantly higher on day 0 and day 1 than with Hygrolife. Absolute humidity measured in chronic obstructive pulmonary disease patients was identical to that measured in the rest of the study population. Tracheal colonization and HME colonization were similar with both HMEs. Bacterial contamination of the ventilator side of both devices was markedly low. CONCLUSIONS: These two purely hygroscopic HMEs provided safe and efficient humidification during a 48-hr period of use in long-term mechanically ventilated medical intensive care unit patients, including chronic obstructive pulmonary disease patients. In addition, they maintained ventilatory circuits clean, despite the absence of filtering media. The cost of mechanical ventilation is consequently reduced. PMID- 12626992 TI - Quantitative analysis of the relationship between sedation and resting energy expenditure in postoperative patients. AB - OBJECTIVE: To analyze quantitatively the relationship between sedation and resting energy expenditure or oxygen consumption in postoperative patients. DESIGN: A prospective, clinical study. SETTING: An eight-bed intensive care unit at a university hospital. PATIENTS: Thirty-two postoperative patients undergoing either esophagectomy or surgery of malignant tumors of the head and neck who required mechanical ventilation and sedation for > or = 2 days postoperatively. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A total of 133 metabolic measurements were performed. Ramsay sedation scale (RSS), body temperature, and the dose of midazolam were evaluated at the time of the metabolic cart study. All patients received analgesia with buprenorphine at a fixed dose of 0.625 microg x kg(-1) x hr(-1) continuously. Midazolam was used for induction and maintenance of intravenous sedation after admission to the intensive care unit. The initial dose was 0.04 mg x kg(-1) x hr(-1) and was adjusted to achieve a desired depth of sedation at 3, 4, or 5 on the RSS every 4 hrs. The degree of sedation was classified into three states: light sedation (RSS 2-3; n = 49), moderate sedation (RSS 4; n = 39), and heavy sedation (RSS 5-6; n = 45). RESULTS: With increasing the depth of sedation, oxygen consumption index (mL x min(-1) x m(-2)), resting energy expenditure index (REEI; kcal x day(-1) x m(-2)), and REE/basal energy expenditure (BEE) decreased significantly. Oxygen consumption index (mean +/- SD), REEI, and REE/BEE were 151 +/- 18, 1032 +/- 120, and 1.29 +/- 0.17 in the light sedation, 139 +/- 22, 947 +/- 143, and 1.20 +/- 0.16 in the moderate sedation, and 125 +/- 16, 865 +/- 105, and 1.13 +/- 0.12 in the heavy sedation, respectively. CONCLUSION: An increase in the depth of sedation progressively decreases in oxygen consumption index and REEI in postoperative patients. PMID- 12626993 TI - Effects of drotrecogin alfa (activated) on organ dysfunction in the PROWESS trial. AB - OBJECTIVE: To assess morbidity in patients with severe sepsis managed with and without drotrecogin alfa (activated). DESIGN: Analysis of secondary end points in a prospective, randomized, double-blind, placebo-controlled, multicenter, phase 3 trial (PROWESS). SETTING: A total of 164 medical institutions in 11 countries. PATIENTS: A total of 1,690 consecutive adult patients with severe sepsis. INTERVENTIONS: A 96-hr infusion of drotrecogin alfa (activated) (human recombinant activated protein C) or placebo. MEASUREMENTS AND MAIN RESULTS: Sequential Organ Failure Assessment (SOFA) scores for cardiovascular, respiratory, renal, hematologic, and hepatic organ systems were measured for 28 days. Mean cardiovascular SOFA scores were significantly lower for patients treated with drotrecogin alfa (activated) compared with placebo patients over this time period (p = .022). Drotrecogin alfa (activated)-treated patients also showed significantly faster resolution of cardiovascular (p = .009) and respiratory (p = .009) dysfunction and significantly slower onset of hematologic organ dysfunction (p = .041) compared with placebo patients for days 1 to 7. No significant differences in morbidity were observed between treatment groups among 28-day survivors. CONCLUSION: Drotrecogin alfa (activated) demonstrated significant improvements in organ function compared with placebo in a large phase 3 clinical trial that has shown a mortality benefit in patients with severe sepsis. PMID- 12626994 TI - Effect of filtration volume of continuous venovenous hemofiltration in the treatment of patients with acute renal failure in intensive care units. AB - OBJECTIVE: We evaluated the variable Kt/V, which has become established in the therapy of end-stage renal disease in acute renal failure, to assess the influence of the filtration volume of continuous venovenous hemofiltration on Kt/V. We measured the variables of acid-base balance and uremia control. DESIGN: Prospective interventional pilot study. SETTING: Medical intensive care unit of a university hospital. PATIENTS: Fifty-six patients with acute renal failure and continuous venovenous hemofiltration treatment. INTERVENTIONS: The patients were consecutively treated with a filtration volume of either 1 L/hr (group 1) or 1.5 L/hr (group 2). MEASUREMENTS AND MAIN RESULTS: Patients with a filtration volume of 1.5 L/hr achieved a Kt/V of 0.8 per day, which was significantly higher than in the patient group treated with 1 L/hr (0.53, p <.05). The filtration volume of 1.5 L/hr led to a markedly better control of blood urea nitrogen concentrations, 69.3 +/- 6.6 mg/dL vs. 52.1 +/- 5.2 (p <.05), and to a much quicker and longer lasting compensation of acidosis. Both groups had acidotic pH at the beginning of therapy (group 1, 7.29 +/- 0.02; group 2, 7.29 +/- 0.02, nonsignificant). In group 2, a significantly higher pH value than in group 1 was measured after 24 hrs of continuous venovenous hemofiltration (p < .001; 7.39 +/- 0.02 vs. 7.31 +/- 0.02). The pH values in group 1 did not normalize until after 4 days. The filtration volume of 1.5 L/hr led to a quicker increase in bicarbonate concentrations after 24 hrs of therapy (group 1, 2.8 +/- 3.2 mmol/L; group 2, 6.5 +/- 3.1 mmol/L, p <.001). CONCLUSIONS: The standardized urea clearance Kt/V is a valuable tool in the treatment of acute renal failure. Higher Kt/V levels were associated with a better control of uremia and acid-base balance. However, there were no differences in the clinical course, patient survival, percentage of patients with or without renal failure who were transferred from the intensive care unit, or Acute Physiology and Chronic Health Evaluation III scores. PMID- 12626995 TI - Improved survival with hospitalists in a pediatric intensive care unit. AB - OBJECTIVE: To compare the survival of patients in a teaching hospital pediatric intensive care unit in which residents provided after-hours in-house coverage with survival in the same unit with hospitalists providing this coverage. DESIGN: Retrospective cohort study. SETTING: Pediatric intensive care units in two teaching hospitals that are managed by the same group of academic pediatric intensivists, one of which transitioned from the traditional resident-staffed model to a hospitalist-staffed model for after-hours in-house coverage. PATIENTS: All pediatric patients admitted to the study pediatric intensive care unit and to the control pediatric intensive care unit from April 1997 through March 1998, the resident era, and from October 1998 through September 1999, the hospitalist era. INTERVENTIONS: Multivariate analysis, with survival as the dependent variable and era (hospitalist vs. resident) as an independent variable, was used to compare odds of survival during the hospitalist era with that of the resident era, adjusted for severity of illness. Multivariate linear regression was used to compare length of stay during the hospitalist era with that of the resident era, adjusted for severity of illness. Pediatric Risk of Mortality scores and those diagnostic categories typically associated with higher mortality rates also were included as independent variables in both analyses to adjust for severity of illness. MEASUREMENTS AND MAIN RESULTS: Multivariate analysis yielded an estimated odds ratio of survival of 2.8 for the hospitalist era compared with the resident era (p = .013), and our analysis supported an independent association between survival and hospitalist era. Multivariate linear regression showed that length of stay, also adjusted for severity of illness, during the hospitalist era was 21.1 hrs shorter than during the resident era (p = .013). Neither survival nor length of stay was significantly associated with era at the control hospital. CONCLUSION: Improved survival with hospitalists, rather than residents, providing after-hours care when an intensivist is not in house suggests that the quality of care of critically ill patients is improved when more experienced physicians are providing bedside care. Shorter length of stay with the hospitalist model also may reflect improved quality of care. PMID- 12626996 TI - Labeled acetate to assess intestinal absorption in critically ill patients. AB - OBJECTIVE: To compare the absorption of carbon-13(13C) acetate-enriched nutrients with D-xylose absorption. DESIGN: Prospective cohort observational study. SETTING: Surgical intensive care unit of a university hospital. PATIENTS: A total of 24 critically ill patients requiring enteral nutritional support. INTERVENTION: The patients were divided into three groups according to the route of 13C acetate administration: 1) gastric, 2) jejunal, and 3) intravenous. D xylose was administered via the same route as enteral nutrition. MEASUREMENTS AND MAIN RESULTS: 13C acetate absorption and oxidation were reflected by pulmonary 13CO2 excretion. Breath 13CO2 isotopic enrichment was measured by mass spectrometry. 13C acetate absorption was rapid, and D-xylose absorption was depressed in all three groups, compared with the normal values (p <.0001). Breath CO isotopic enrichment was similar after intravenous and jejunal administration but slightly delayed during the first 240 mins after gastric administration (p <.01). Enteral feeding was well tolerated: mean energy delivery amounted to 77%, 88%, and 86% of measured resting energy expenditure on days 1-3. CONCLUSIONS: Gastric and jejunal 13C acetate are rapidly absorbed in critically ill surgical patients requiring enteral nutrition, contrasting with a depressed or delayed D xylose absorption. 13CO2 recovery kinetics was similar after jejunal or intravenous 13C acetate and slightly depressed after gastric administration. Further studies are required to determine the value of labeled nutrients to assess gastric emptying and intestinal absorption. PMID- 12626997 TI - Hospital mortality rate and length of stay in patients admitted at night to the intensive care unit. AB - OBJECTIVE: Although admission of patients to a medical ward after 5:00 pm has been associated with increased mortality rate and possibly shorter hospital stay, the association between timing of admission to the intensive care unit and outcome has not been studied. The objective of this study was to determine whether there are any associations between the timing of patient admission to a medical intensive care unit and hospital outcome. DESIGN: A retrospective cohort study that used an Acute Physiology and Chronic Health Evaluation III database containing prospectively collected demographic, clinical, and outcome information for patients. Patients were divided according to the time of admission into daytime (from 7:00 am to 5:00 pm) and nighttime admissions. We further subdivided nighttime admissions into two groups (regular and heavy workload) according to the number of patients who were admitted during the same shift. SETTING: Medical intensive care unit (a 15-bed unit in an academic referral hospital). PATIENTS: 6,034 patients consecutively admitted to our medical intensive care unit over a 5 yr period starting April 10, 1995. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The patients admitted at night had a lower mortality rate (13.9 vs. 17.2%, p < .0001), adjusted for admission source and severity of illness. Their hospital stay was shorter, 11.0 days +/- 13.5 (median 7) vs. 12.7 +/- 14.8 (median 8; p < .0001), as was their intensive care unit stay, 3.5 +/- 4.4 days (median 2) vs. 3.9 +/- 4.7 (median 2; p < .0001), compared with the daytime admission group. The nighttime shifts that admitted three or more patients (heavy workload) had the same mortality rate (13.2%) as those with fewer admissions (14.5%; p = .5961). Hospital and intensive care unit stays were also similar in both workload groups. CONCLUSIONS: Nighttime admission to our intensive care unit is not associated with a higher mortality rate or a longer hospital or intensive care unit stay compared with daytime admission. PMID- 12626998 TI - Effect of prostacyclin on platelets, polymorphonuclear cells, and heterotypic cell aggregation during hemofiltration. AB - OBJECTIVES: Hemodialysis activates both platelets and leukocytes, which play a role in the development of multiple organ dysfunctions in critically ill patients. Prostacyclin inhibits both cell types. To examine the hypothesis that prostacyclin prevents cellular activation during clinical hemofiltration, we investigated the expression of activation markers on platelets and leukocytes using whole blood flow cytometry. DESIGN: Prospective, randomized, double-blind, controlled trial. SETTING: Intensive care unit. PATIENTS: A total of 24 consecutive, critically ill, mechanically ventilated patients with acute renal failure secondary to sepsis or major surgery. INTERVENTIONS: For anticoagulation during hemofiltration, patients received either unfractionated heparin or unfractionated heparin and prostacyclin (5 ng x kg(-1) x min(-1)). Anticoagulants were administered into the extracorporeal circuit before the hemofilter. Blood samples were obtained from an arterial catheter before hemofiltration and from the inlet and outlet lines of the extracorporeal circuit at 1 and 24 hrs during hemofiltration. MEASUREMENTS AND MAIN RESULTS: Expression of GP IIb-IIIa and P selectin on adenosine diphosphate-activated platelets and platelet-leukocyte aggregation were significantly lower after the passage of blood through the hemofilter in patients receiving an extracorporeal infusion of prostacyclin plus heparin when compared with control patients receiving heparin only. There were no statistically significant differences in the expression of CD11b on leukocytes between the two groups. CONCLUSIONS: These findings suggest that prostacyclin reversibly inhibits platelet function by diminishing the expression of platelet fibrinogen receptors and P-selectin and reduces heterotypic platelet-leukocyte aggregation during clinical hemofiltration. However, prostacyclin fails to inhibit leukocyte activation at clinically relevant doses. PMID- 12626999 TI - Chronic alcohol abuse is associated with an increased incidence of acute respiratory distress syndrome and severity of multiple organ dysfunction in patients with septic shock. AB - OBJECTIVE: Alcohol is one of the most commonly used drugs in the world and causes dysfunction in many vital organs. However, the effects of chronic alcohol abuse on acute lung injury and nonpulmonary organ dysfunction are relatively unexplored. The goal of this study was to determine the effects of chronic alcohol abuse on the incidence and severity of the acute respiratory distress syndrome and multiple organ dysfunction syndrome in patients with septic shock. DESIGN: Multicenter prospective epidemiologic study. SETTING: Intensive care units of four university urban hospitals. PATIENTS A total of 220 critically ill patients with septic shock. MEASUREMENTS AND MAIN FINDINGS: Thirty percent of the patients (66 of 220) were identified as having a history of chronic alcohol abuse based on a positive response to an alcohol screening questionnaire. The incidence of acute respiratory distress syndrome in patients with a positive history of chronic alcohol abuse was 70% (46 of 66), compared with 31% (47 of 154) in individuals without a history of chronic alcohol abuse (p < .001). After adjusting for differences in the source of infection, sex, age, chronic hepatic dysfunction, diabetes, severity of illness, nutritional status, and smoking status, the effects of chronic alcohol abuse on the incidence of acute respiratory distress syndrome remained significant (p < .001; odds ratio, 3.70; 95% confidence interval, 1.83-7.71). The effect of the source of infection (pulmonary vs. nonpulmonary) on the development of acute respiratory distress syndrome also remained significant in this multivariable analysis (p < .001; odds ratio, 3.68; 95% confidence interval, 1.95-7.18). Based on the highest daily Sequential Organ Failure Assessment score, patients with a history of chronic alcohol abuse had more severe nonpulmonary organ dysfunction when compared with nonalcoholics (9.42 +/- 3.89 vs. 8.05 +/- 4.10, p = .01). After adjusting for source of infection, sex, age, nutritional status, history of diabetes, and smoking status, the effects of chronic alcohol abuse on the incidence of nonpulmonary organ dysfunction also remained significant (p = .03; odds ratio, 2.07; 95% confidence interval, 1.09-3.97). CONCLUSIONS: We conclude that chronic alcohol abuse is an independent risk factor for acute respiratory distress syndrome and increases the severity of nonpulmonary organ dysfunction in patients with septic shock. PMID- 12627001 TI - Lethal systemic capillary leak syndrome associated with severe ventilator-induced lung injury: an experimental study. AB - OBJECTIVE: We report the evolution of severe ventilator-induced lung injury associated with lethal systemic capillary leak syndrome, when sheep were ventilated at a peak inspiratory pressure of 50 cm H2O, at a respiratory rate of 8 breaths.min, with an inspiratory time of 2.5 secs. DESIGN: A prospective laboratory animal study. SETTING: Experimental animal research laboratory. SUBJECTS: Mixed breed sheep. INTERVENTIONS: Sheep were anesthetized, paralyzed, and mechanically ventilated. MEASUREMENTS AND MAIN RESULTS: This sheep model was characterized by a rapidly evolving massive anasarca, hemoconcentration, cardiac dysfunction, multiple system organ failure, and severe ventilator-induced lung injury. Cardiovascular changes and profound hemoconcentration developed within 6 hrs from the start of mechanical ventilation, along with a major decline in pulmonary compliance and deterioration in arterial blood gases. When total static lung compliance decreased to 0.15 mL (cm H2O)(-1) x kg(-1) (7-30 hrs), the sheep were randomized to two groups. Group I received high (recruitive) positive end expiratory pressure (9-20 cm H2O), adjusted as needed; group II received low (supportive) positive end-expiratory pressure (2-6 cm H2O). Sheep in both groups progressively deteriorated and died with cardiocirculatory failure and multiple system organ failure within 12-24 hrs from start of treatment. CONCLUSIONS: This model of lethal systemic capillary leak syndrome with multiple system organ failure differs greatly from our previous sheep model of acute ventilator-induced lung injury in which sheep were ventilated with a peak inspiratory pressure of 50 cm H2O, a respiratory rate of 4 breaths x min(-1), and an inspiratory time of 1.35 secs, without inducing capillary leak syndrome. The mere change of respiratory rate from 4 to 8 breaths x min(-1), with a near doubling of the inspiratory time to 2.5 secs, although maintaining eucapnia, resulted in lethal systemic capillary leak syndrome and multiple system organ failure with both gross and microscopic pathology of lungs greatly different from our previous model of mechanical ventilation-induced acute respiratory distress syndrome. PMID- 12627000 TI - Helium-oxygen versus air-oxygen noninvasive pressure support in decompensated chronic obstructive disease: A prospective, multicenter study. AB - OBJECTIVE: To study whether noninvasive pressure support ventilation (NIPSV) with helium/oxygen (He/oxygen), which can reduce dyspnea, PaCO2, and work of breathing more than NIPSV with air/oxygen in decompensated chronic obstructive pulmonary disease, could have beneficial consequences on outcome and hospitalization costs. DESIGN: Prospective, randomized, multicenter study. SETTING: Intensive care units of three tertiary care university hospitals. PATIENTS: All patients with chronic obstructive pulmonary disease admitted to the intensive care units for NIPSV during a 24-month period. INTERVENTIONS: Patients were randomized to NIPSV with air/oxygen or He/oxygen. NIPSV settings, number of daily trials, decision to intubate, and intensive care unit and hospital discharge criteria followed standard practice guidelines. RESULTS: A total of 123 patients (male/female ratio, 71:52; age, 71 +/- 10 yrs, Acute Physiology and Chronic Health Evaluation II, 17 +/- 4) were included. Intubation rate (air/oxygen 20% vs. He/oxygen 13%) and length of stay in the intensive care unit (air/oxygen 6.2 +/- 5.6 vs. He/oxygen 5.1 +/- 4 days) were comparable. The post-intensive care unit hospital stay was lower with He/oxygen (air/oxygen 19 +/- 12 vs. He/oxygen 13 +/- 6 days, p < .002). Cost of NIPSV gases was higher with He/oxygen, but total hospitalization costs were lower by $3,348 per patient with He/oxygen. No complications were associated with the use of He/oxygen. CONCLUSION: He/oxygen did not significantly reduce intubation rate or intensive care unit stay, but hospital stay was shorter and total costs were lower. He/oxygen NIPSV can be safely administered and could prove to be a cost-effective strategy. PMID- 12627002 TI - Protective role of heme oxygenase-1 in the intestinal tissue injury in an experimental model of sepsis. AB - OBJECTIVE: The aim of this study was to examine the role of heme oxygenase-1 induction in the intestinal tissue injury in a rat model of sepsis. DESIGN: Randomized, masked, controlled animal study. SETTING: University-based animal research facility. SUBJECTS: Sprague-Dawley male rats, weighing 220-250 g (n = 126). INTERVENTIONS: Rats were injected with lipopolysaccharide (10 mg/kg) intraperitoneally. Another group of rats was injected with interleukin-6 (10 microg/kg) intravenously. In some rats, tin mesoporphyrin (1 micromol/kg) was administered intravenously 1 hr before lipopolysaccharide treatment. MEASUREMENTS AND MAIN RESULTS: Following lipopolysaccharide treatment, expression of heme oxygenase-1 and nonspecific delta-aminolevulinate synthase (ALAS-N), the rate limiting enzymes of heme catabolism and biosynthesis, respectively, was examined in various regions of the intestine. Lipopolysaccharide treatment markedly increased heme oxygenase-1 messenger RNA and protein concentrations in the mucosal epithelial cells in the duodenum and the jejunum, whereas its expression in the ileum and the colon was hardly detectable and was not influenced by the treatment. ALAS-N messenger RNA was also more markedly increased in the duodenum, the jejunum, and the ileum than in the colon following lipopolysaccharide treatment. Interleukin-6 administration also induced heme oxygenase-1 and ALAS-N gene expression in a pattern similar to that following lipopolysaccharide treatment. In contrast to the marked heme oxygenase-1 expression in the upper intestine, lipopolysaccharide-induced mucosal injury and inflammation in the upper intestine were far less than observed in the lower intestine as judged both by tumor necrosis factor-alpha gene expression and by histologic analysis. Of note, inhibition of heme oxygenase activity by tin mesoporphyrin produced a significant tissue injury in the upper intestine of the lipopolysaccharide treated animals. CONCLUSIONS: Intestinal heme oxygenase-1 and ALAS-N gene expression was regulated in a site-specific manner in a rat model of sepsis. Our findings also suggest that heme oxygenase-1 induction may play a fundamental role in protecting mucosal epithelial cells of the intestine from oxidative damages that occur in sepsis. PMID- 12627003 TI - Early multiple organ failure after recurrent endotoxemia in the presence of vasoconstrictor-masked hypovolemia. AB - OBJECTIVE: Critically ill patients who develop multiple organ failure during systemic inflammatory states are often predisposed to hypovolemia and vasoconstrictor therapy. Although numerous investigations have evaluated the sequelae of systemic inflammation, no data are available on the contribution of chronic vasoconstrictor-masked hypovolemia to organ dysfunction and morphology. DESIGN: Prospective, randomized laboratory investigation. SETTING: University research laboratory. SUBJECTS: Eighteen adult chronically instrumented sheep. INTERVENTIONS: The animals were randomly assigned to one of three groups. In the norfenefrine-masked hypovolemia plus endotoxemia (NMH+ENDO) group, mean arterial pressures of 80 mm Hg were maintained by using the alpha1-adrenergic catecholamine norfenefrine for 52 hrs during hypovolemia. Hypovolemia was induced by hemorrhage (about 23 mL x kg(-1)) until mean arterial pressures reached 40 mm Hg. Endotoxin (0.5 microg x k(-1)) was then injected after 4, 16, 28, and 40 hrs. The NMH group received norfenefrine-masked hypovolemia but no endotoxin. In the ENDO group, recurrent endotoxemia was induced during normovolemia. MEASUREMENTS AND MAIN RESULTS: Despite profound differences in fluid management, cardiovascular filling pressures were not statistically different between groups. Endotoxemia induced norfenefrine-refractory shock (p < .05 vs. the other groups) and contributed to renal dysfunction only during vasoconstrictor-masked hypovolemia. Norfenefrine-masked hypovolemia caused disseminated cardiac cell necrosis independent of endotoxemia (p < .05 vs. ENDO). CONCLUSIONS: Hypovolemia can be masked when volume status is monitored by filling pressures. In this new model of endotoxemia-associated multiple organ failure, chronic vasoconstrictor masked hypovolemia turned systemic inflammation into a life-threatening condition with renal and cardiovascular failure. Cardiomyocyte necroses were caused by vasoconstrictor-masked hypovolemia but were unrelated to cardiovascular failure. PMID- 12627004 TI - Interleukin-1 beta rapidly inhibits aortic endothelium-dependent relaxation by a DNA transcription-dependent mechanism. AB - OBJECTIVES: This study examined the effects of interleukin-1 beta on isometric tension development and relaxation in isolated rat aortic rings in response to the alpha-1 adrenergic agonist phenylephrine, the endothelium-dependent vasodilator acetylcholine, and the endothelium-independent vasodilator sodium nitroprusside. DESIGN: Randomized, controlled, paired design. SETTING: Animal laboratory within a university department of physiology. SUBJECTS Paired aortic thoracic aortic rings from specific pathogen-free Sprague-Dawley rats. INTERVENTIONS: Series I examined the potential for interleukin-1 beta to cause early arterial endothelial dysfunction. Paired aortic rings were incubated for 2 hrs with interleukin-1 beta or vehicle. Series II examined the potential for inhibition of DNA transcription to attenuate interleukin-1 beta-mediated endothelial dysfunction. Paired rings received either dactinomycin or vehicle before interleukin-1 beta incubation. Series III quantified the degree to which inhibition of DNA transcription inhibited early interleukin-1 beta-mediated endothelial dysfunction. Paired rings received either dactinomycin pretreatment followed by interleukin-1 beta incubation, or pretreatment and incubation with inert vehicles. Series IV assessed the effects of interleukin-1 beta on responsiveness to an exogenous nitric oxide donor, sodium nitroprusside, in the presence of the nitric oxide synthesis inhibitor N omega-nitro-L-arginine methyl ester. MEASUREMENTS AND MAIN RESULTS: Incubation with interleukin-1 beta for 2 hrs had no effect on contractile response but attenuated endothelium-dependent relaxation significantly relative to control. Dactinomycin pretreatment inhibited early interleukin-1 beta-mediated endothelial dysfunction. The combination of interleukin-1 beta and dactinomycin produced effects on endothelium-dependent relaxation that were not different from that seen in rings not exposed to interleukin-1 beta. Interleukin-1 beta attenuated responsiveness to sodium nitroprusside relative to control. CONCLUSIONS: Interleukin-1 beta causes an early impairment of endothelium-dependent vasorelaxation with an onset that precedes its effects on systemic contractility. This impairment occurs via a mechanism that is wholly or predominantly dependent on DNA transcription. The altered vasorelaxation induced by interleukin-1 beta is at least partly mediated by a reduction in nitric oxide responsiveness. PMID- 12627005 TI - Tracheobronchial injury during intratracheal pulmonary ventilation in rabbits. AB - OBJECTIVE: We compared tracheobronchial injury following short-term intratracheal pulmonary ventilation (ITPV) and conventional mechanical ventilation (CMV) in a healthy rabbit model. ITPV, a form of tracheal gas insufflation, has been shown to decrease deadspace ventilation and increase CO2 removal and therefore may reduce ventilator-induced lung injury. SETTING: Medical center laboratory. SUBJECTS: Twenty-five rabbits. INTERVENTIONS: Rabbits were randomly assigned to either ITPV or CMV (n = 15 and 10, respectively). Both groups were mechanically ventilated for 8 hrs at the same ventilator settings (FIO2, 0.4; rate, 30 breaths/min; flow, 4 L x min(-1); positive end-expiratory pressure, 4 cm H2O; tidal volume, 40 mL). Peak, mean, and end-expiratory carinal pressures, ITPV flow rate, and hemodynamic variables were continuously monitored. Tissue samples for histologic analysis were obtained postmortem from the trachea contiguous to the tip of the endotracheal tube, the distal trachea, the carina, and the main bronchus. The histologic sections were scored, in a single-blind fashion, for ciliary damage, ulceration, hemorrhage, overall inflammation, intraepithelial inflammatory infiltrate, and edema. MEASUREMENTS AND MAIN RESULTS: ITPV was associated with significantly lower Paco and deadspace ventilation ratio than CMV. The combined tracheobronchial injury scores for all samples were significantly higher in the ITPV group compared with the CMV group (p <.005; Mann Whitney U test). The ITPV injury scores, compared with CMV injury scores, were significantly higher at the carina and main bronchus (p <.01; Kruskal-Wallis test followed by Dunn's multiple comparison test). The area adjacent to the endotracheal tube showed the same degree of damage in both groups. Analysis of the injury scores in individual damage categories demonstrated the greatest difference in the ulceration category (p <.001). CONCLUSIONS: In our study, ITPV, compared with CMV at the same minute ventilation, was associated with a significantly greater difference in tracheobronchial damage at the carina and main bronchus. We postulate that this difference may have been caused by the turbulence of the gas flow generated by the small-caliber ITPV catheter used in our neonatal-size animal model. PMID- 12627006 TI - Endotoxin binding to erythrocyte membrane and erythrocyte deformability in human sepsis and in vitro. AB - OBJECTIVE: Several studies have shown that lipopolysaccharide and lipid A impair red blood cell deformability and. However, it is unclear whether impaired red blood cell deformability is associated with binding of lipopolysaccharide to the red blood cell membrane. DESIGN: Analysis of hydroxymyristic acid content in red blood cell membranes and red blood cell deformation in patients with Gram negative septicemia and after incubation of red blood cells from healthy adults with 100 microg of lipid A or 1 mg of lipopolysaccharide per milliliter of red blood cell in buffer solution and in whole blood. Hydroxymyristic acid is a fatty acid of the lipid A part of lipopolysaccharide in most Gram-negative bacteria. SETTING: University research laboratories. SUBJECTS: Ten healthy adults and four patients with clinical and laboratory signs of septicemia. INTERVENTIONS: Blood sampling. MEASUREMENTS AND MAIN RESULTS: Red blood cell deformation was measured with a laser-diffraction shearing device (Rheodyn) and a computerized micropore filtration system (CTA). Lipopolysaccharide and lipid A binding to red blood cell membranes was studied by measuring the amide-linked hydroxymyristic acid by gas chromatography. The detection rates of hydroxymyristic acid were 82% for lipopolysaccharide and 79% for lipid A in buffer solution. In membranes of washed red blood cell, the detection rates of lipopolysaccharide and lipid A were 0.26 +/- 0.03% (2.6 +/- 0.3 microg/mL) and 1.3 +/- 0.5% (1.3 +/- 0.5 microg/mL), and in red blood cell membranes of whole blood the detection rates were 2.6% (25.5 microg/mL) and 4.1% (4.1 microg/mL), respectively. The lipopolysaccharide content in red blood cell membranes of septic patients ranged from 47 to 103 microg/mL of red blood cell. Red blood cell deformation in the Rheodyn and in the CTA were not influenced by lipopolysaccharide incubated with washed red blood cells. In the Rheodyn, red blood cell deformation was significantly decreased by 18% after lipid A incubation in washed red blood cells, by 26% after lipopolysaccharide incubation in whole blood, and by 31% in septic patients. Similar effects were observed when we used the CTA. CONCLUSIONS: Red blood cell deformation is decreased in septic patients, after incubation of washed red blood cells with lipid A and of whole blood with lipopolysaccharide. Lipopolysaccharide did not influence red blood cell deformation after incubation with washed red blood cells. The decrease of red blood cell deformation was related to the amount of hydroxymyristic acid measured in red blood cell membranes, suggesting that endotoxin binding directly affects mechanical properties of red blood cells. PMID- 12627007 TI - Effect of anti-CD14 monoclonal antibody on clearance of Escherichia coli bacteremia and endotoxemia. AB - OBJECTIVE: To determine the effects of an anti-CD14 monoclonal antibody on the clearance of a bacteremic Escherichia coli challenge in the presence or absence of antimicrobial agents. DESIGN: Prospective randomized animal study. SETTING: University-affiliated research laboratory. SUBJECTS: New Zealand White rabbits weighing 1.5-2.5 kg. INTERVENTIONS: Animals were pretreated with either an anti lapine CD14 monoclonal antibody (immunoglobulin G2a, 5 mg/kg intravenously) or an isotype control monoclonal antibody. The animals then were challenged with 1 x 10(6) E. coli 018:K1 in the presence or absence of ceftazidime (50 mg/kg intravenously). There were four groups of six animals randomized to receive either anti-CD14 monoclonal antibody without ceftazidime, isotype control monoclonal antibody without ceftazidime, anti-CD14 monoclonal antibody with ceftazidime, or isotype control antibody with ceftazidime. MEASUREMENTS AND MAIN RESULTS: Serial measurement of quantitative bacteremia and endotoxemia was performed over 24 hrs after the administration of the bacterial challenge. Animals also underwent necropsy with quantitative bacterial cultures from multiple organ tissue samples. The anti-lapine CD14 monoclonal antibody significantly impaired the bloodstream clearance of E. coli (p <.01) and increased quantitative counts of E. coli in tissue culture samples when compared with isotype control antibody in the absence of simultaneous administration of ceftazidime. No differences in quantitative bacteremia, endotoxemia, or organ tissue counts were found after anti-CD14 antibody and control antibody-treated animals in the presence of ceftazidime treatment. CONCLUSIONS: Anti-CD14 monoclonal antibody has the capacity to interfere with the innate immune response and systemic microbial clearance in experimental animals with E. coli bacteremia. The concomitant administration of effective antimicrobial therapy eliminated differences in the rate of microbial clearance between the control antibody and the CD14 monoclonal antibody. These results indicate that care should be taken in clinical trials with anti-CD14 monoclonal antibodies to ensure that adequate antimicrobial therapy is administered in the presence of systemic bacterial infection. PMID- 12627008 TI - Correlation between transcranial interleukin-6 gradient and outcome in patients with acute brain injury. AB - OBJECTIVE: This study was performed to examine both brain and systemic interleukin-6 (IL-6) release in patients with an acute brain injury (ABI), to study whether a correlation exists between the transcranial IL-6 gradient during the first days after injury and prognosis, and finally, to investigate the relationship between a nucleotide polymorphism at position -174 in the promoter of the gene encoding IL-6, IL-6 responsiveness, and clinical evolution. DESIGN: Prospective clinical investigation. SETTING: A 19-bed intensive care unit in a university hospital. PATIENTS AND METHODS: A total of 62 patients were followed up for 3 days after acute brain injury, and both their arterial and jugular IL-6 levels were measured serially and at the moment of brain death diagnosis. Genetic polymorphism of IL-6 was also determined in all patients. Data were correlated with those from score procedures for clinical severity. Neurologic outcome was graded according to the Glasgow Outcome Scale 6 months after injury. IL-6 levels and IL-6 genotyping was performed in control healthy individuals. MAIN RESULTS: There is a significant transcranial IL-6 gradient at admission and at the moment of brain death. The gradient is higher in those patients who evolved toward a fatal outcome during the first 6 months after injury (p <.001). There is significant correlation between the transcranial IL-6 gradient and the acute brain injury severity. CONCLUSIONS: IL-6 is elevated in patients with acute brain injury, and a significant relationship exits between the severity of acute brain injury and the transcranial IL-6 gradient at admission. It can be considered to be a prognosis marker at admission. When data at the moment of brain death are considered, venous IL-6 (p <.01) and the transcranial IL-6 gradient (p <.005) are significantly higher than at the time of admission. Although the IL-6 C allele is associated with significantly lower concentrations of IL-6, there was no correlation between low or high IL-6 responders and patient outcome. PMID- 12627009 TI - S-100beta protein-serum levels in healthy children and its association with outcome in pediatric traumatic brain injury. AB - OBJECTIVE: To describe normal serum levels of S-100beta in healthy children and determine whether serum S-100beta levels after traumatic brain injury are associated with outcome. DESIGN: Prospective cohort study. SETTING: Urban, tertiary care, children's teaching hospital. PATIENTS: A total of 136 healthy children and 27 children with traumatic brain injury. METHODS: Serum S-100beta levels were measured in 136 healthy children. A total of 27 children with traumatic brain injury had S-100beta levels collected within 12 hrs of injury. Other indices of severity of injury measured were admission Glasgow Coma Scale score, and Pediatric Risk of Mortality score at 24 hrs (PRISM 24). Outcome was measured by the Pediatric Cerebral Performance Category (PCPC) score at hospital discharge and 6 months postinjury or at death. MEASUREMENTS AND MAIN RESULTS: S 100beta levels in healthy children had a mean of 0.3 microg/L (90% confidence interval, 0.03-1.47) and inversely correlated with age, (r = -.32, p <.001). In children with traumatic brain injury, 6-month postinjury outcome inversely correlated with Glasgow Coma Scale score (r = -.47, p =.01) and correlated with PRISM 24 score (r =.83, p <.001) and S-100beta levels (r =.75, p <.001). Six months postinjury, comparing good outcome (PCPC < or = 3, n = 20) vs. poor outcome (PCPC > or = 4, n = 7), median admission Glasgow Coma Scale scores were 8 (range, 3-15) and 3 (range, 3-7, p =.01), median PRISM 24 scores were 7 (range, 0 19) and 30 (range, 18-35, p <.001), and median S-100beta levels were 0.85 microg/L (range, 0.08-4.8 microg/L) and 3.6 microg/L (range, 1.4-20 microg/L, p <.001), respectively. A serum S-100beta level of > or =2.0 microg/L is associated with poor outcome, with a sensitivity of 86% and a specificity of 95%. The area under the receiver operating curve for S-100beta was 0.94 (+/-0.05). CONCLUSIONS: Serum S-100beta levels in healthy children have a moderate inverse correlation with age. After traumatic brain injury in children, the acute assessment of serum S-100beta levels seems to be associated with outcome. PMID- 12627010 TI - Cardiovascular management of septic shock. PMID- 12627011 TI - Discrepant attitudes about teamwork among critical care nurses and physicians. AB - OBJECTIVE: To measure and compare critical care physicians' and nurses' attitudes about teamwork. DESIGN: Cross-sectional surveys. SETTING: Eight nonsurgical intensive care units in two teaching and four nonteaching hospitals in the Houston, TX, metropolitan area. SUBJECTS: Physicians and nurses who worked in the intensive care units. MEASUREMENTS AND MAIN RESULTS: Three hundred twenty subjects (90 physicians and 230 nurses) responded to the survey. The response rate was 58% (40% for physicians and 71% for nurses). Only 33% of nurses rated the quality of collaboration and communication with the physicians as high or very high. In contrast, 73% of physicians rated collaboration and communication with nurses as high or very high. By using factor analysis, we developed a seven item teamwork scale. Multivariate analysis of variance of the items yielded an omnibus ( [7, 163] = 8.37; p <.001), indicating that physicians and nurses perceive their teamwork climate differently. Analysis of individual items revealed that relative to physicians, nurses reported that it is difficult to speak up, disagreements are not appropriately resolved, more input into decision making is needed, and nurse input is not well received. CONCLUSIONS: Critical care physicians and nurses have discrepant attitudes about the teamwork they experience with each other. As evidenced by individual item content, this discrepancy includes suboptimal conflict resolution and interpersonal communication skills. These findings may be the result of the differences in status/authority, responsibilities, gender, training, and nursing and physician cultures. PMID- 12627012 TI - Predictive value of somatosensory evoked potentials for awakening from coma. AB - OBJECTIVES: A systematic review of somatosensory evoked potentials performed early after onset of coma, to predict the likelihood of nonawakening. The pooled results were evaluated for rates of awakening, confidence intervals, and the possibility of rare exceptions. DATA SOURCES: Forty-one articles reporting somatosensory evoked potentials in comatose patients and subsequent outcomes, from 1983 to 2000. STUDY SELECTION: Studies were included if they reported coma etiology, age group, presence or absence of somatosensory evoked potentials, and coma outcomes. DATA EXTRACTION: We separated patients into four groups: adults with hypoxic-ischemic encephalopathy, adults with intracranial hemorrhage, adults and adolescents with traumatic brain injury, and children and adolescents with any etiologies. Somatosensory evoked potentials were categorized as normal, abnormal, or bilaterally absent. Outcomes were categorized as persistent vegetative state or death vs. awakening. DATA SYNTHESIS: For each somatosensory evoked potential result, rates of awakening (95% confidence interval) were calculated: adult hypoxic-ischemic encephalopathy: absent 0% (0%-1%), abnormal 22% (17%-26%), normal 52% (48%-56%); adult intracranial hemorrhage: absent 1% (0% 4%), present 38% (27%-48%); adult-teen traumatic brain injury: absent 5% (2%-7%), abnormal 70% (64%-75%), normal 89% (85%-92%); child-teen: absent 7% (4%-10%), abnormal 69% (61%-77%), normal 86% (80%-92%). CONCLUSIONS: Somatosensory evoked potential results predict the likelihood of nonawakening from coma with a high level of certainty. Adults in coma from hypoxic-ischemic encephalopathy with absent somatosensory evoked potential responses have <1% chance of awakening. PMID- 12627013 TI - Sepsis: a landscape from the emergency department to the intensive care unit. PMID- 12627014 TI - Treatment of ventilator-associated pneumonia: get it right from the start. PMID- 12627015 TI - Cardiovascular dysfunction of sepsis: a nitric oxide- and l-arginine-deficient state? PMID- 12627016 TI - Bacterial colonization of the respiratory tract under artificial ventilation: is there proof of clinically relevant effects of the endotracheal tube orientation under positioning therapy? PMID- 12627017 TI - Mechanical ventilation: weapon of mass destruction or tool for liberation? PMID- 12627018 TI - Is prediction imperative before prevention? PMID- 12627019 TI - Percutaneous extracorporeal life support: reserve for patients with reversible causes of shock and cardiac arrest. PMID- 12627020 TI - Predicting mortality in acute respiratory distress syndrome: circulatory system knows best. PMID- 12627021 TI - Sulfite-induced propofol oxidation: a cause for radical concern. PMID- 12627022 TI - Sublingual capnometry: useful gadget or just another toy? PMID- 12627023 TI - Benefit of recombinant human activated protein C beyond 28-day mortality: there is more to life than death. PMID- 12627024 TI - Pediatric hospitalists improve critical care outcomes. PMID- 12627025 TI - A time for work and a time for rest. PMID- 12627026 TI - Continuous dialysis as systemic therapy in the critically ill patient? PMID- 12627027 TI - Noninvasive ventilation with helium-oxygen rather than air-oxygen in acute exacerbations of chronic obstructive disease? PMID- 12627028 TI - Sepsis and hypovolemia: two bad. PMID- 12627029 TI - Teamwork and collaboration in critical care: lessons from the cockpit. PMID- 12627030 TI - Clinical neurophysiologic assessment of comatose patients. PMID- 12627031 TI - Abnormal response to adrenocorticotropic hormone tests in critically ill human immunodeficiency virus-infected patients: are we really dealing with adrenal insufficiency? PMID- 12627032 TI - Children should be allowed to dictate the pace of their recovery. PMID- 12627033 TI - Bispectral Index in the intensive care setting. PMID- 12627034 TI - The hepatosplanchnic area is not a common source of lactate production. PMID- 12627035 TI - Does oral contraceptive use lead to hormone therapy use in women doctors? Our bodies, our choices, our practice. PMID- 12627036 TI - Raloxifene to prevent cardiovascular disease? Don't use it yet! PMID- 12627037 TI - Role of progestogen in hormone therapy for postmenopausal women: position statement of The North American Menopause Society. AB - OBJECTIVE: To create an evidence-based position statement regarding the role of progestogen in postmenopausal hormone therapy (estrogen plus a progestogen, or EPT) for the management of menopause-related symptoms. DESIGN: NAMS followed the general principles established for evidence-based guidelines to create this document. Clinicians and researchers acknowledged to be experts in the field of postmenopausal hormone therapy were enlisted to review the evidence obtained from the medical literature and develop a position statement for approval by the NAMS Board of Trustees. RESULTS: The primary role of progestogen in postmenopausal hormone therapy is endometrial protection. Unopposed estrogen therapy (ET) is associated with a significantly increased risk of endometrial hyperplasia and adenocarcinoma. Adding the appropriate dose and duration of progestogen to ET has been shown to lower that risk to the level found in never-users of ET. The clinical goal of progestogen in EPT is to provide endometrial protection while maintaining estrogen benefits and minimizing progestogen-induced side effects, particularly uterine bleeding. EPT discontinuance correlates with uterine bleeding-women with more days of amenorrhea have higher rates of continuance. All US Food and Drug Administration-approved progestogen formulations will provide endometrial protection if the dose and duration are adequate. Progestogens may diminish the beneficial effects of ET on cardiovascular risk factors. However, no EPT (or ET) regimen should be initiated for the primary or secondary prevention of cardiovascular heart disease. Some progestogens may negatively affect mood. Adding progestogen to ET does not decrease the breast cancer risk, although it does not seem to increase mortality. Progestogen increases mammographic density, which is reversed after discontinuation of use. Progestogen has limited effect on the bone-enhancing action of ET. In general, the side effects of added progestogen are mild, although they may be severe in a small percentage of women. CONCLUSIONS: Progestogen should be added to ET for all postmenopausal women with an intact uterus to prevent the elevated risk of estrogen-induced endometrial hyperplasia and adenocarcinoma. There is no consensus on a preferred regimen for all women. By changing the progestogen type, route, or regimen, clinicians can individualize therapy to minimize side effects, especially uterine bleeding, and limit any effects on ET benefits while providing adequate endometrial protection. PMID- 12627038 TI - Clinical and personal relationships between oral contraceptive and hormone replacement therapy use among US women physicians. AB - OBJECTIVE: To examine relationships between a history of oral contraceptive (OC) use and current use of or intention to use hormone replacement therapy (HRT). DESIGN: The Women Physicians' Health Study examined a stratified random sample of US women MDs, aged 30 to 70 years (4,501 respondents; a 59% response rate). RESULTS: Among postmenopausal physicians who previously used OCs, current HRT use was significantly associated ( < 0.05) with being younger, living somewhere besides the East Coast, being sexually active, being an obstetrician/gynecologist, having no history of breast cancer, having a longer use of OCs, and being posthysterectomy. Among premenopausal physicians, intended future HRT use was significantly associated with being white, being an obstetrician/gynecologist, being in good health, living somewhere besides the East Coast, being a longer user of OCs, and having more extensive, recent continuing medical education. Among women who had taken both OCs and HRT, there were no significantly elevated rates in any of the 15 health conditions we examined (after controlling for family history). Postmenopausal physicians who took HRT (and premenopausal OC-using physicians intending to take HRT) were significantly more likely to counsel their patients on HRT use. Among post-OC HRT users, 44% counseled their postmenopausal female patients on HRT at least yearly versus 22% of post-OC HRT-nonusers (74% v 45% among such primary care physicians). CONCLUSIONS: Physicians' personal OC/HRT use may strongly affect their patient counseling practices. HRT use after OC use was not associated with any obvious increases in examined diseases in this population. PMID- 12627039 TI - Raloxifene is a better antioxidant of low-density lipoprotein than estradiol or tamoxifen in postmenopausal women in vitro. AB - OBJECTIVE: The oxidation of low-density lipoprotein (LDL) is an important factor in the development of atherosclerosis. The antioxidant activity of some compounds buffers the free radicals generated either endogenously or exogenously, thus decreasing the potential damage mediated by oxidation. Estrogens are potent antioxidants of LDL, in vitro and in vivo, a mechanism that could probably influence the cardioprotection associated with hormone replacement therapy in postmenopause. We conducted an in vitro study of the antioxidant effect on LDL of two selective estrogen receptor modulators, raloxifene (RLX) and tamoxifen (TMX), comparing them with the known antioxidant effect of estradiol (E2 ). DESIGN: LDL was isolated by ultracentrifugation from plasma obtained from 12 healthy, untreated, postmenopausal women. Aliquots containing 0.5 mg of LDL protein were incubated for 4 h with CuSO4 (15 micro M) to induce oxidative stress and with one of the three compounds studied: RLX, TMX, or E2 at doses of 0, 1, 2, 3, 5, 15, 50, and 500 micro M, and 1 and 2 mM. Malonaldehyde (MDA, nmol/mg protein) was measured as a marker of LDL oxidation. RESULTS: E2 induced a dose-dependent decrease in MDA concentration. MDA values decreased significantly, as compared with baseline, starting at a concentration of 2 micro M for RLX and 3 micro M for both, TMX, or E2. The dose necessary to reduce the generation of MDA by 50% was significantly lower for RLX (3.3 micro M, < 0.001) than for E2 (24.6 micro M ) or TMX (35.3 micro M). The area under the curve also showed a higher antioxidant activity for RLX compared with TMX or E2 ( < 0.001). CONCLUSIONS: The in vitro antioxidant activity of RLX is substantially more potent than TMX or E2. This finding, added to the other beneficial effects of the drug in the cardiovascular system, could imply some cardioprotector effect. PMID- 12627040 TI - Soy protein and isoflavone effects on vasomotor symptoms in peri- and postmenopausal women: the Soy Estrogen Alternative Study. AB - OBJECTIVE: To investigate the efficacy of dietary soy proteins containing differing amounts of isoflavones on the number and severity of vasomotor symptoms (hot flashes and night sweats) in peri- and postmenopausal women. DESIGN: A double-masked, randomized, controlled, clinical trial was conducted. A total of 241 community-dwelling women reporting vasomotor symptoms at baseline were randomized into one of three groups. In all groups, participants consumed a daily supplement containing 25 g of soy protein and were randomly assigned to one of three groups: (a) isoflavone extracted soy protein (control), (b) soy protein with a medium dose of isoflavones (42 mg/day), or (c) soy protein with a higher dose of isoflavones (58 mg/day). The primary outcome measure in this trial was change in reported vasomotor symptoms. RESULTS: A reduction in the number and severity of vasomotor symptoms was observed in all three treatment groups. No significant differences in the number and severity of vasomotor symptoms were observed among the high isoflavone, middle isoflavone, or control groups. The lack of a between-treatment group effect was observed even after stratified by number of baseline symptoms and use of traditional hormone replacement therapy. CONCLUSIONS: These data suggest that soy protein containing 42 or 58 mg of isoflavones is no more effective than isoflavone-extracted soy protein for improving the number and severity of vasomotor symptoms in peri- and postmenopausal women. PMID- 12627041 TI - The early response of the postmenopausal endometrium to tamoxifen: expression of estrogen receptors, progesterone receptors, and Ki-67 antigen. AB - OBJECTIVE: To enlighten the early response of endometrium to tamoxifen by assessing the expression of estrogen receptors, progesterone receptors, Ki-67, and the histological response in endometria from normal postmenopausal women treated for 21 days with tamoxifen. DESIGN: A total of 40 women, scheduled to undergo vaginal hysterectomy because of uterine prolapse, were randomly assigned to the tamoxifen group (20 mg/day; 20 women) or the control group (20 women). Samples were obtained from the upper and the lower thirds of the uterine cavity. Standard immunohistochemical staining of estrogen and progesterone receptors and of Ki-67 was performed on frozen sections. Staining was assessed using semiquantitative immunoreactivity scores. RESULTS: Simple endometrial hyperplasia was diagnosed in 18 of the 20 samples exposed to tamoxifen compared with only 2 of the 20 controls ( P< 0.0005). Staining was increased in both the epithelium and stroma in the tamoxifen samples, a difference that was significant for estrogen receptors in glandular epithelium (180 +/- 80 v 110 +/- 110; P< 0.05). Also, Ki-67 antigen was expressed more frequently in both glandular epithelium ( P< 0.05) and stroma ( P< 0.05) in the tamoxifen samples. CONCLUSIONS: Tamoxifen rapidly up-regulated the cell proliferation markers, an effect that was associated with enhanced growth as confirmed by increased expression of estrogen receptors and Ki-67, in addition to a high incidence of glandular hyperplasia. PMID- 12627042 TI - Effects of bilateral ovariectomy and postoperative hormonal replacement therapy with 17beta-estradiol or raloxifene on serum leptin levels. AB - OBJECTIVE: To verify the effects of hypoestrogenism and replacement therapy on body mass index (BMI) and leptin in ovariectomized women. DESIGN: We conducted a longitudinal study on 56 women undergoing abdominal bilateral ovariectomy divided into three groups: 19 untreated controls, 18 scheduled to receive 17beta estradiol, and 19 on raloxifene starting 15 days after surgery. Height, weight, and BMI were recorded on the day of surgery, 5 and 15 days later, and then after 6 months. Leptin and estradiol concentrations were determined by radioimmunoassay on the day of surgery, days 1, 5, and 15, and 6 months after. RESULTS: Leptin levels rose significantly on the day after surgery [median (range): 18.2 (9.8 25.0), 12.5 (9.1-20.9), and 20.5 (12.9-24.5); P< 0.01 v basal] and returned to values similar to baseline on day 5 in all groups. Six months later, controls showed significantly higher leptin levels in comparison with both treated women and basal values [median (range): 19.7 (10.4-22.8), 11.0 (7.6-16.9), and 13.5 (9.1-14.8) ng/ml; P< 0.01). Estradiol levels decreased in all groups, reaching statistical significance 5 days after surgery ( P< 0.01 v basal). A significant rise was observed 6 months after surgery in women treated with estrogens ( P< 0.01). Six months after surgery, BMI increased in untreated controls in comparison with treated women and baseline, although not significantly. CONCLUSIONS: The absence of modifications in leptin on days 5 and 15 after ovariectomy suggests that, in humans, estrogens may not exert an important effect on leptin secretion. After 6 months, replacement therapy maintained leptin levels and BMI was unmodified, whereas untreated controls showed a significant increase in leptin and a trend toward higher BMI, suggesting that replacement therapy may prevent changes in fat distribution and in leptin levels. PMID- 12627043 TI - Effect of estrogen replacement therapy on parathyroid hormone secretion in elderly postmenopausal women. AB - OBJECTIVE: Women undergo two phases of involutional bone loss that have opposing effects on parathyroid hormone (PTH) secretion. During the early phase, the loss of the direct restraining effect of estrogen on bone resorption causes an outflow of skeletal calcium into the extracellular fluid. This causes a compensatory decrease in PTH secretion. In the late phase, loss of extraskeletal effects of estrogen (on intestinal and renal calcium handling) leads to increases in whole body losses of calcium and a compensatory increase in PTH secretion. Moreover, long-term estrogen replacement therapy (ERT) suppresses both basal and stimulated PTH secretion in these women. Whereas the effects of estrogen on PTH secretion have been assumed to be due to its extraskeletal actions, estrogen may also have direct effects on the parathyroid glands. The goal of the present study was to test for these possible direct effects of estrogen on PTH secretion. DESIGN: Basal and ethylenediaminetetraacetic acid (EDTA)-stimulated PTH secretion was assessed in 10 elderly postmenopausal women (mean age, 76.4 years) before and after acute (3 days) estrogen replacement with transdermal estradiol, 0.1 mg/day. In addition, similar studies were performed in 10 age-matched women (mean age, 74.5 years) who had been on long-term ERT. These women were studied before and after 3 days of estrogen withdrawal. RESULTS: Estrogen treatment or withdrawal had no significant effect on either basal or stimulated PTH secretion. CONCLUSIONS: These data provide evidence that, in elderly postmenopausal women, estrogen does not have significant direct effects on PTH secretion and point to the importance of the actions of estrogen on intestinal and renal calcium handling as the major mechanisms for its effects on modulating calcium homeostasis and, indirectly, PTH secretion. PMID- 12627044 TI - Comparison of the difference in histopathology and cell cycle kinetics among the postmenopausal endometrium treated with different progestins in sequential combined hormone replacement therapy. AB - OBJECTIVE: To investigate the difference in histopathology and cell cycle kinetics in the menopausal endometrium treated with sequential-combined hormone replacement therapy (HRT) using different types and doses of progestins. DESIGN: A randomized, double-blind, 1-year study was conducted. In a menopause clinic of a university hospital, 241 postmenopausal women using HRT were included for the study of histopathology and cell cycle analysis. Conjugated equine estrogens, 0.625mg/day, were administered for 25 days (days 1-25) of each month, and the following were also administered for 14 days (days 12-25): in group A ( n= 102), medroxyprogesterone acetate (MPA), 5 mg/day; in group B ( n= 66), MPA, 10mg/day; and in group C ( n= 73), dydrogesterone, 20mg/day. Endometrial sampling was performed after at least 10 months of treatment. Fifty-two premenopausal women were also enrolled for the comparative studies (group Y). The S-G2-M fractions in the cell cycle were used as the marker of proliferation. RESULTS: Most menopausal endometria were normal regardless of the regimens of HRT. Endometrial hyperplasia was only found in two cases (both in group A). The S-G2-M fractions of the endometrial cells in all three menopausal groups showed no statistically significant difference. It appeared that S-G2-M fractions increased from normal postmenopausal to normal premenopausal endometria to postmenopausal hyperplasia to premenopausal hyperplasia. The S-G2-M fractions of the normal menopausal endometrial cells were lower than those of the premenopausal controls either in normal or in hyperplastic categories. CONCLUSIONS: Our study showed that there is no difference between the effect of MPA and dydrogesterone used in sequential combined HRT based on the cycle kinetics of the menopausal endometrium. PMID- 12627045 TI - Menopausal transition and psychological development. AB - OBJECTIVE: To study women's psychological development during menopause and to examine the relationship between women's appraisal of menopause and symptom reporting. DESIGN: A population-based sample of 130 healthy women was assessed annually for 5 consecutive years using semistructured interviews, a menopausal symptom rating scale, and the Symptom Checklist-90 rating scale. RESULTS: Qualitative analyses of the interviews showed that the majority of the women (57%) had neutral beliefs about menopause, whereas 31% were pessimistic and 12% were optimistic. Optimistic and neutral expectations were associated with low levels of symptom reporting, whereas the pessimistic appraisal was significantly related to elevated symptom scores. The majority of the neutral and pessimistic women reappraised menopause during the study period, and at the last follow-up, 67% appraised menopause positively. A positive reappraisal among the initially pessimistic women was associated with more frequent statements about personal growth compared with the other subgroups. CONCLUSIONS: Menopause has a developmental potential and is a positive period for most women. PMID- 12627047 TI - The effects of stressful life events, coping, and cortisol on HIV infection. AB - What is the role of stress and coping in changes in immunologic and clinical indicators of human immunodeficiency virus disease progression? There is substantial evidence that stressful life events and passive coping strategies, such as denial, may have a detrimental effect on HIV disease progression. Given the harmful effects of stress and passive coping, the author reviews the limited research testing the efficacy of interventions, such as cognitive-behavioral therapies for HIV-infected persons. Finally, in trying to understand psychoimmune relationships in HIV, the evidence is examined for the mediating and direct effects of cortisol, a hormone associated with stress, on HIV disease progression. Delineating the role of psychosocial factors and cortisol on HIV disease progression may aid in the development of new interventions for this devastating disease. PMID- 12627048 TI - Stress management and psychoneuroimmunology in HIV infection. AB - Does stress management affect psychological and immune functioning in persons with human immunodeficiency virus infections? Stress-management techniques, such as relaxation training and imagery, cognitive restructuring, coping-skills training, and interpersonal-skills training, may reduce anxiety, depression, and social isolation in HIV-infected persons by lowering physical tension and increasing a sense of control and self-efficacy. A psychoneuroimmunologic model is proposed wherein these psychological changes are hypothesized to be accompanied by an improved ability to regulate neuroendocrine functioning, which in turn may be associated with a partial normalization of immune system functions such as lymphocyte proliferation and cytotoxicity, providing more efficient surveillance of latent viruses that may contribute directly to increased HIV replication and generate opportunistic infections or cancer if left unchecked. Such a normalization of stress-associated immune system decrements are hypothesized to forestall or minimize increases in viral load and expression of clinical symptoms. This model is useful for testing the factors contributing to the health effects of stress-management interventions in HIV-infected persons. In this context, one general research strategy for testing the effects of stress management interventions is to target them toward the more prevalent psychosocial challenges that HIV-infected people face at various points in the disease process; enroll an HIV-infected population (eg, HIV-positive homosexual and bisexual men) into a randomized trial; and monitor changes in cognitive, affective, behavioral, and social factors in parallel with hormonal, immunologic, viral, and clinical changes over the course of time. This article will review the major psychoneuroimmunologic findings that have emerged using this paradigm and suggest future research directions and clinical applications. PMID- 12627049 TI - Depression and HIV infection: impact on immune function and disease progression. AB - Can psychological factors, such as depression, affect human immunodeficiency virus progression? HIV infection is viewed as a chronic illness in which those infected often confront a number of emotional challenges and physical health and disease-related issues. Over the past 20 years, there has been increasing evidence that depression and other mood-related disturbances are commonly observed among HIV-positive individuals. There is also mounting data showing that depressive symptoms might further impact upon specific elements of immune system functioning and influence quality of life and health status. This paper will highlight studies examining the prevalence of depression during HIV infection and review some of the evidence examining the impact of depressive symptoms on immune function and HIV disease progression. PMID- 12627050 TI - Neuropsychopharmacologic treatment of depression and other neuropsychiatric disorders in HIV-infected individuals. AB - How can neuropsychiatric disorders and syndromes be underdiagnosed and inadequately treated in individuals infected with human immunodeficiency virus? Depression in particular is among the most prevalent diagnoses and there is a solid foundation of data from controlled clinical studies that has begun to examine the efficacy of various antidepressants in HIV-infected persons. This article summarizes essential findings pertaining to the use of psychotropic medications to treat depression and other neuropsychiatric disorders in the context of immunodeficiency. This includes discussion of clinically significant treatment considerations (eg, efficacy, side effects, drug-drug interactions) derived from the existing literature. Taken together, there is compelling evidence that psychopharmacologic intervention can improve the quality of life of mentally ill HIV-infected individuals. PMID- 12627065 TI - Myocardial extracellular signal regulatory kinases are activated by laser treatment. AB - AIM: Transmyocardial laser revascularization (TMR) and percutaneous transluminal myocardial laser revascularization (PTMR) have anti-anginal effects on certain groups of patients with ischemic heart disease, possibly by inducing myocardial neoangiogenesis through the mechanical injury. Here we examine the effects of TMR and PTMR on extracellular signal regulatory kinases (ERKs), which have been implicated in the control of neoangiogenesis in vitro. METHODS: Eight pigs were anesthetized with ketamine and fentanyl. In five pigs TMR was performed in the left ventricular anterior wall and PTMR in the posterior wall. Biopsies were taken either after 2 hours, 3, 7, 14, and 30 days. Three pigs served as controls and provided samples for baseline values and time-matched controls at 2 hours and 3 days. ERKs activity was determined by increased phosphorylation of myelin basic protein. Total ERKs protein abundance was determined by Western blot with an antibody against ERK1 and ERK2. RESULTS: It was found that ERKs activity was higher in all samples from the laser treated myocardium than in the control sample at baseline (TMR: >or=1878 pmol Pi x min(-1) mg pr(-1) vs 104 pmol Pi x min(-1) mg pr(-1), respectively, p<0.05. PTMR: >or=346 pmol Pi x min(-1) mg pr( 1) vs 136 pmol Pi min(-1) mg pr(-1), respectively, p<0.05). The time-matched samples showed increased activities of ERKs after laser treatment. The protein level of ERKs in myocardium treated with TMR and PTMR parallelled the data on ERKs activity. CONCLUSIONS: Our data suggest that the ERKs system is activated in the porcine heart by the mechanical injury of TMR and PTMR. PMID- 12627066 TI - A randomized study of combining maze surgery for atrial fibrillation with mitral valve surgery. AB - AIM: Mitral valve surgery seldom suppresses atrial fibrillation (AF), present prior to surgery. Maze III surgery eliminates AF in >80% of cases, the reason why combining this procedure with mitral valve surgery in patients with AF seems worthwhile. We prospectively studied the outcome of combining the Maze III procedure with mitral valve surgery. METHODS: Thirty-five patients with AF and a mean age of 64 years undergoing mitral valve surgery were prospectively randomized according to a 2.5:1 ratio to surgery with (n=25), or without (n=10) maze III and followed for at least 1 year. RESULTS: At discharge and after 12 months freedom from AF was 56% and 92%, respectively, in the maze group, and 0% and 20%, respectively, in patients without maze (group differences at discharge p=0.002, after 12 months p=0.0007). Sinus node incompetence was seen in 1 of 25 maze patients requiring pacing. No in-hospital or late death occurred; stroke was observed in 1 patient (without maze). Quality of life markedly improved after surgery, but did not differ between patients with or without maze surgery. CONCLUSIONS: This first prospective randomized study shows that combining maze III with mitral valve surgery resulted in a significantly better elimination of preoperative AF than mitral valve surgery alone. As the quality of life did not differ between patients with, or without maze surgery, additional maze surgery is primarily recommended in patients in whom anticoagulation therapy can be avoided after surgery, specifically in patients with scheduled mitral valve plasty. PMID- 12627067 TI - Midterm results of conservative repair of the incompetent bicuspid aortic valve. AB - AIM: Repair of the bicuspid aortic valve is not generally considered the treatment of choice. Our success with this procedure leads us to report our immediate and mid-term RESULTS: METHODS: From August 1993 to December 2000, 19 patients with aortic regurgitation due to congenital aortic valve underwent aortic valve repair (17 men and 2 women with a mean age of 42+/-17 years; range, 16 to 70 years). The mean preoperative aortic regurgitation grade was 3.1+/-0.8 on a scale of 1 to 4. Mean preoperative New York Heart Association functional class was 1.9+/-0.8. Fourteen patients had pure aortic regurgitation, 2 also had infectious endocarditis, 1 had angina pectoris, and 2 had an ascending aortic aneurysm. RESULTS: There was 1 hospital death (5.2%), and 1 patient required re operation due to recurrent infectious endocarditis. Mean aortic regurgitation grade at discharge was 1.1+/-0.9, and functional class was 1.1+/-0.2. All patients were followed for a mean duration of 40+/-23 months (range, 0.5 to 84 months). There was 1 late death, and two patients required aortic valve replacement. The 5-year survival rate was 90+/-7%. The 1- and 5-year re-operation free rates were 87+/-12% and 76+/-23%. CONCLUSIONS: Bicuspid aortic valve repair is a safe procedure with good early postoperative RESULTS: However midterm results are not satisfactory. Re-operation is a promising alternative and progress aortic regurgitation were complications. Bicuspid aortic valve repair to valve replacement that requires additional study to individualize treatment. PMID- 12627068 TI - Initial clinical and hemodynamic experience with Edwards MIRA mechanical bileaflet valve. AB - AIM: An enhanced bileaflet valve, the Edwards MIRA feminine Mechanical Valve became available in 1998. Favorable hydrodynamic features and a redesigned sewing ring encouraged us to implant this device in indicated patients. Hemodynamics and clinical performance parameters were evaluated. METHODS: Between February 1998 and October 1999, 338 patients (171 males, 167 females) underwent native valve replacement with a MIRA prosthesis. Mean age 56.6+/-13.6 years, 320 patients were in NYHA class III/IV. Sixty-seven patients had echocardiographic examinations. Standard cardiopulmonary bypass was employed utilizing institutionally accepted implantation techniques. Aortic valve replacement was performed in 163 patients, mitral valve replacement in 134 patients, 35* double valve replacements and 1 triple valve replacement. RESULTS: Follow-up is 98% complete. Mean follow-up is 6.9+/-3.3 months (178.2 patient years). There were no operative deaths. Four early deaths were seen (1.18%). Late deaths reported in 12 patients. Linearized rate of late mortality was 6.7% per patient year (ppy). Overall actuarial survival at 13 months is 92.2%. Mean gradients and Effective Orifice Areas (EOA's) are comparable to other bileaflet valves. Linearized rates for valve related complications was 4.49% ppy. Only 5 transient thromboembolic events (TE = 2.81% ppy) and 3 non-structural valve dysfunction events (NSVD = 1.68%) were seen. No reports of bleeding events, prosthetic endocarditis, valve thrombosis or structural valve deterioration. One patient required mitral valve reoperation for perivalvular leak. CONCLUSIONS: Short-term hemodynamic and clinical results are comparable to other bileaflet valves. The sewing ring is non-obstructive, compliant with smoother needle penetration. Early clinical results are encouraging, follow-up should be continued. PMID- 12627069 TI - Elevated levels of s-100beta correlate with neurocognitive outcome after cardiac surgery. AB - AIM: S-100beta is a specific astroglial protein whose serum level increases after cerebral injury. The purpose of this study was to investigate the correlation between elevated levels of S-100beta and the neurocognitive outcome after cardiac surgery. METHODS: Fifty consecutive patients undergoing elective coronary artery bypass grafting were studied. Serum S-100beta levels were measured on induction of anaesthesia, at the 15(th) minute, at skin closure and on the 1(st) postoperative day. Neurocognitive outcome was evaluated by STAI-T and Zung tests preoperatively and by Mini-mental state examination every postoperative day until discharge. Neurocognitive tests and S-100beta levels were correlated within the scope of risk factors by Pearson correlation. RESULTS: Serum S-100beta was not detected preoperatively. Peak serum S-100beta levels were reached at skin closure in 36 of 50 patients (72%). In 24 hours, serum S-100beta disappeared in 25 patients but was still elevated in 11 (22%). A highly significant correlation was demonstrated between the duration of CPB and peak serum S-100beta levels (r=0.91). There was a weak correlation between age and peak S-100beta levels (r=0.62). Nine patients (18%) had a positive MMSE test which correlated well with persistent high serum S-100 levels (r=0.98). CONCLUSIONS: Serum S-100beta is a promising early biochemical marker for cerebral injury following cardiac surgery within a good correlation with the CPB time, age and especially with neurocognitive tests. PMID- 12627070 TI - Role of tetrahydrobiopterin on ischemia-reperfusion injury in isolated perfused rat hearts. AB - AIM: It has recently been shown that nitric oxide synthase in the presence of suboptimal levels of tetrahydrobiopterin (BH(4)), an essential cofactor of nitric oxide synthase, may favor increased production of oxygen free radicals. This study was designed to define the role of BH(4) in myocardial ischemia-reperfusion injury. METHODS: Isolated perfused rat hearts were subjected to 37 degrees C ischemia and reperfusion. Hearts were received with BH(4) or vehicle for 5 min just before ischemia and during the first 5 min of the reperfusion period. The effects of BH(4) on left ventricular function, myocardial contents of lipid peroxidation and high energy phosphates, and levels of lactate dehydrogenase and nitrite plus nitrate in perfusate before ischemia and after reperfusion were estimated. Moreover, the effect of BH(4) given with 2,4-diamino-6 hydroxypyrimidine (DAHP), a selective inhibitor of BH(4) production, intraperitoneally 24 h before the experiments were estimated. RESULTS: BH(4) improved contractile and metabolic abnormalities in reperfused hearts. Furthermore, BH(4) significantly alleviated ischemic contracture during ischemia, and restored diminished perfusate levels of nitrite plus nitrate after reperfusion. On the other hand, DAHP-treatment aggravated ischemia-reperfusion induced functional and metabolic abnormalities. Administration of BH(4) improved DAHP-induced functional and metabolic abnormalities. CONCLUSION: Results demonstrated that BH(4) lessens ischemia-reperfusion injury in isolated perfused rat hearts. Conversely, deficiency of BH(4) seems to accelerate endothelial dysfunction and myocardial ischemia-reperfusion injury. Present data may be compatible with the hypothesis that nitric oxide synthase in the presence of insufficiency of BH(4) serve as the cause of oxidative injury. PMID- 12627071 TI - Does direct visualization of peripatch areas in beating heart eliminate the risk of residual ventricular septal defect in adult patients? AB - AIM: Although the incidence of residual ventricular septal defect (VSD) following surgical therapy in the modern series is very low, especially the risk of hemodynamically insignificant shunt still exists. Intraoperative assessment of residual shunt is useful in identifying patients at risk of having subsequently required reoperation and reintervention for residual VSD before chest closure. METHODS: In 87 patients who were operated because of isolated VSD (Group I), VSD was closed under cardioplegic arrest and right atriotomy or right ventriculotomy were closed in the beating heart after aortic cross-clamp removal. The VSD patch was watched out for residual shunt and additional sutures were placed if it existed. Results of this technique have been compared with the other 216 (Group II) in which all procedures of the VSD closure were performed under cardioplegic arrest. Transosephageal echocardiography (TEE) was performed for evidence of residual shunting intraoperatively and postoperatively in all patients. RESULTS: In group I, additional sutures were placed for residual shunt in 14 patients (16.1%), and insignificant residual shunt was detected in only one (1.1%) patient at early postoperative period (p<0.05, according to group II). In group II, there was hemodynamically insignificant residual shunt in 31 patients (14.5%), and 9 patients (4.2%) were reoperated for significant shunt (p<0.05). CONCLUSION: Transatrial or transventricular inspection to peripatch areas in the beating heart is a safe technique to detect a residual shunt, an observation that may eliminate reoperation. PMID- 12627072 TI - Off-pump coronary artery bypass grafting in a patient with AIDS, acute myocardial infarction, and severe left main coronary artery disease. AB - A 48-year-old male patient with AIDS presented with postinfarct unstable angina, decreased left ventricular function (EF 35%), significant left main coronary artery disease, and total occlusion of the proximal left anterior descending and right coronary arteries. In order to avoid the potential immunosuppressive effect of cardiopulmonary bypass (CPB) in an already compromised host with an already low CD4+ helper/inducer T cell count (180/microL) and high retroviral load (165,000 copies/mL), the application of beating-heart technology and off-pump coronary bypass grafting was an ideal indication. The patient underwent successfully off-pump/CPB coronary revascularization. The triple drug combination of highly active antiretroviral therapy (HAART) was resumed postoperatively. The patient was discharged from the hospital on the 7(th) postoperative day. The CD4+ count was 142/microL and the viral load decreased to 450 copies/mL. Seven months post-operatively the patient was free of angina and without shortness of breath. The CD4+ count was 160/(m)L and the viral load undetectable. Improved survival of HIV positive patients has resulted in a shift from caring for terminally ill patients to caring for patients with chronic illness. While protease inhibitors have positively affected survival, they may also cause plasma lipid abnormalities, which can lead to severe premature coronary artery disease. Therefore, an increasing population of AIDS and HIV positive patients with coronary artery disease may require cardiac interventions in the near future. Coronary revascularization without CPB and its potential immunocompromising effect may play an important role in patients with severe coronary artery disease and AIDS. PMID- 12627074 TI - Partial anomalous left pulmonary venous return. AB - We report the case of a 52-year-old woman with partial left anomalous pulmonary venous return who was successfully treated surgically without the use of extracorporeal circulation by diverting the anomalous left vertical vein into the left atrium through a postero-lateral thoracotomy. PMID- 12627073 TI - Mitral stenosis due to fibrous tissue overgrowth after mitral valve repair. AB - We report an extremely rare case of fibrous tissue overgrowth 3 years after mitral valve repair using a mitral annuloplasty ring in a 53-year-old woman who underwent mitral valve replacement for mitral stenosis. Whitish fibrous tissue had overgrown from the ring on the atrial side of the annulus, and had severely reduced the valvular area. However, the motion of the mitral leaflets was not restricted. Considering the presence of concomitant aortitis syndrome, it is strongly suggested that the overgrowth of fibrous tissue was promoted as a reaction to chronic inflammation. PMID- 12627075 TI - Right atrial rupture following a hoof kick to the chest wall. AB - We report a case of right atrial rupture due to blunt trauma by a hoof kick to the anterior chest wall. We describe our successful management of this case. We suggest that a diagnosis of cardiac chamber rupture should be considered in all cases of blunt thoracic trauma. PMID- 12627076 TI - Effect of thrombus on abdominal aortic aneurysm wall dilation and stress. AB - AIM: Our goal is to understand how a mural thrombus may influence the pressure transmitted to and the dilation experienced by the abdominal aortic aneurysm (AAA) wall. METHODS: Two intact AAAs with mural thrombus were removed from patients and pressurized to 100 mmHg. The pressure was measured using a micro-tip needle transducer inserted in the aneurysm wall and advanced through the thrombus. In 1 patient with AAA, similar measurements were made in vivo. Also, in vitro, in the two aneurysms the dilation as a function of pressure was measured using the markers on the surface before and after the thrombus was removed. RESULTS: Both, in vitro and in vivo, in the presence of the thrombus the pressure transmitted to the aneurysm wall was 91+/-10% of luminal pressure and at 6 mm from the wall it was 96+/-5%. The aneurysm dilated more in the pressure range of 0-40 mmHg (2-8%) than in the range of 40-100 mmHg (0.4-1.8%). Upon removal of the thrombus these dilations increased significantly to 4-15% and 0.9-3.3%, respectively. Overall, the strains (dilation) in the circumferential and longitudinal directions were similar before the thrombus was removed. CONCLUSIONS: Even though the thrombus allows the transmission of luminal pressure to the aneurysm wall, it may prevent the aneurysm from rupture by diminishing the strain on the wall. Consistent with this, a mechanical model of the thrombus proposed is "a thrombus as a fibrous network adherent to the aneurysm wall". PMID- 12627078 TI - Surgical management of chronic total occlusion of abdominal aorta. AB - AIM: The surgical management of the totally occluded abdominal aorta is highly complex and possible complications are more likely to be seen. We reviewed our experience to make a base for future endoluminal procedures. METHODS: Seventy-one patients with totally occluded abdominal aorta were operated between 1985 and 1998 in a main referral hospital of the social security organization and the results of vascular interventions were evaluated retrospectively. Juxtarenal aortic occlusion was seen in 52% of these patients. All the patients were operated using the transperitoneal approach and adequate dissection to control renal arteries. Limited thrombectomy through infrarenal aortotomy without transecting the aorta was done and continued with standard aortic graft insertion except for 1 patient with porcelain aorta. RESULTS: Follow-up was 76.9+/-41.9 (SD) months. Fourteen patients had concurrent femoro-distal bypass and 2 patients had concurrent renal bypass. Perioperative mortality was 5.6% and 26.5% of patients needed a second intervention during follow-up. Five-, 10- and 13-year survival and freedom from secondary operation was as follows: 84%, 56% and 44%, 81%, 54% and 42%. Cumulative primary and secondary graft patencies at 5- and 10 years were 68%, 63% and 92%, 92%, respectively. CONCLUSIONS: Surgical intervention is beneficial for patients with totally occluded aorta even if ischemic complaints are relatively mild and stable. Acceptable mortality rates and long-term results form a basis for future endovascular interventions. PMID- 12627077 TI - Rationale of the surgical treatment of carotid kinking. AB - AIM: Elongation and tortuosity of the internal carotid artery (ICAET) is a common angiographic, angioMR or Duplex scanning finding: it can be "pure" and, in a great majority of cases, it is not correlated to neurological symptoms. It can be associated with atherosclerotic bifurcation plaque, therefore in this case, indications to surgery follow that of carotid stenosis. On the other hand in some patients ICAET seems potentially correlated to hemispheric or non hemispheric symptoms: ICAET may show as kinking with a wide or narrow acute angle, single (< shaped) or double (Z shaped), or less frequently as a coiling (S,U, or C shaped). Surgical indications are controversial. In the author's opinion, surgery may represent the safest tool in the prevention of a stroke due to carotid occlusion, in selected patients. The aim of this study is to describe the author's experience in the surgical treatment of carotid kinking not associated with significant atherosclerotic lesions. METHODS: From March 1994 to March 2001, 29 patients (11 male, 18 female) with a pure ICAET underwent surgery. Patients presented hemispheric symptoms (24.13%), non hemispheric symptoms (41.3%) or both (27.5 %). Two asymptomatic patients (6.9%) underwent surgery because of contralateral carotid occlusion. RESULTS: The postoperative (within 30 days from operation) results, no mortality was observed, 1 patient presented a stroke (3.4%), and 1 patient had a TIA at awakening (negative cerebral CT scan). All patients with hemispheric symptoms (15 patients) had complete remission, whereas only 6 out of 12 patients (50%) presenting non-hemispheric symptoms had remission (1 patient underwent a controlateral ICAET correction). CONCLUSIONS: The natural history of symptomatic and asymptomatic ICAET is practically unknown, but in some cases selected indication to surgery is justified. Surgery was indicated for patients with transient ischaemic attacks ( hemispheric symptoms); in asymptomatic patients presenting a kinking with an angle inferior to 30 degrees, and a contralateral carotid artery occlusion; in patients with non hemispheric symptoms, after a screening to exclude all other possible neurological or non neurological causes with duplex scan positive for significant increase of flow velocity in ICA and positive cerebral CT scan or MR scan for ischaemic lesions in the homolateral hemisphere, and/or a flow inversion in anterior cerebral artery or flow reduction in the middle cerebral artery, according to different head positions (rotation and flex-extension). PMID- 12627079 TI - A rare vascular malformation, Klippel-Trenaunay syndrome. Report of a case with deep vein agenesis and review of the literature. AB - The Klippel-Trenaunay syndrome (KTS) is a rare anomaly characterized by naevus, soft tissue hypertrophy and varicosities which is mainly unknown etiology. Associated deep venous system abnormalities have been reported with KTS. Here, a 4-year-old-boy with KTS was reported, because of the associated popliteal vein, common femoral vein with its deep and superficial branches, and external iliac vein agenesis. The originality of the presented case is that we have never observed such a case before. PMID- 12627080 TI - Bronchoscopic lung volume reduction. AB - AIM: Because traditional lung volume reduction involves major surgery in unfit patients, resource implications are formidable. Many patients are too ill for consideration and overall survival rates have lead to questions about its continuation. We report on a new method of lung volume reduction, through a bronchoscopic approach. Patients with severe dyspnoea with end stage emphysema were recruited. Target areas for collapse therapy were upper lobes or segments with severe destruction, occupying a large volume, with surrounding lung compression and little perfusion. These were delineated radiologically and with ventilation-perfusion scanning. Endobronchial bloc-kade was initially with detachable, silicone balloons and later with especially designed and hospital manufactured stainless steel wire stents containing bio-compatible sponge. METHODS: Five males and 3 females with preoperative breathlessness at rest and muscle wasting were treated. Operative time was a mean of 67.5 min. Rehabilitation began immediately and 7 patients left hospital the following day. RESULTS: Five patients had improvements in well-being, dyspnoea, exercise tolerance, lifestyles and medication requirements. One patient survived at least 2 years and 2 are still alive 4(1/2) years later. Endoscopic balloon replacement with wire and sponge devices was required in 5 patients. Complications were related to intraoperative oxygenation problems in 2 patients and late infections in 4. CONCLUSIONS: Bronchoscopic lung volume reduction for the treatment of end stage emphysema subjectively improved the majority of patients. The use of stents was promising: balloons were disappointing. PMID- 12627081 TI - Experimental use of an albumin-glutaraldehyde tissue adhesive for sealing tracheal anastomoses. AB - AIM: The purpose of this study was to test the performance of an albumin glutaraldehyde tissue adhesive (BioGlue, manufactured by CryoLife Inc., Kennesaw, GA, USA) when used on tracheal resections in rabbits, which is a sensitive model to investigate the biocompatibility of the glue. METHODS: The 24 animals were anesthetized and underwent cervicotomy with resection of a 10 mm long tracheal segment. The experimental group (18 animals with 2, 4 and 12 week endpoints) had a tracheal anastomosis performed with a maximum of 4 sutures for the approximation of the tissue margins. The anastomotic line was then circumferentially covered with the adhesive. Control animals (6 animals, 4 week endpoint) had a tracheal anastomosis performed with the use of twice interrupted, airtight running suture. The experiments were conducted after approval by the Institutional Ethics Committee and in accordance with the European Convention on Animal Care. RESULTS: Macroscopic evaluation revealed a tight closure of the anastomosis in 23 animals. One rabbit developed tracheo-cutaneous fistula, 2 rabbits experienced intraluminal granulations due to infection, and 1 rabbit developed tracheal stenosis due to insufficient sutures with axis-displacement of the anastomosed tracheal lumina. On microscopic examination, after 2 weeks an inflammatory tissue response consisting of neutrophils, macrophages and foreign body giant cells was found surrounding the glued area. After 4 weeks the tissue was granulomatous in character with an increasing number of multinucleated giant cells. In general, persistent granulomatous inflammation and fibrous scar tissue was seen after 12 weeks. Both, macroscopically and microscopically, fibroangioblastic tissue responses were found in the control group after 4 weeks. CONCLUSIONS: Despite secondary healing disruptions such as granuloma formation, our investigations suggest that the results of albumin-glutaraldehyde tissue adhesive sealed tracheal anastomoses with a few approximating sutures are comparable with those using suture technique. Short term results demonstrated good biocompatibility of the glue. PMID- 12627082 TI - Video-assisted thoracoscopic lung biopsy in the diagnosis of diffuse interstitial lung disease. A prospective study. AB - AIM: Video-assisted thoracoscopic lung biopsy is commonly performed for diagnosis of diffuse interstitial lung disease. This study reports our experience with this technique in terms of diagnostic accuracy, complications and appropriate site for biopsy. METHODS: A prospective non-randomized study. SETTING: Thoracic Surgery Department, Chest Diseases Hospital, Kuwait. PATIENTS AND INTERVENTION: 79 consecutive patients undergoing thoracoscopic lung biopsy for diffuse interstitial lung disease from January 1995 to December 2001. Patients were divided into 2 groups; 34 patients with single biopsy from lingula or right middle lobe (RML) and 45 with 2 biopsies from other sites. MEASURES: comparison of operative and postoperative data, diagnostic accuracy, and complications between the two groups. RESULTS: The mean age of the patients was 38.9 years (range 15-75 years). There was no difference in the groups in operative time, 24 hours postoperative pleural drainage, chest tube drainage days, and hospital stay. Histologic diagnosis was achieved in 76/79 patients (96%). The diagnostic yield of each group was comparable (32/34 from lingula/RML; 44/45 other sites), (p=0.3). Postoperative complications occurred in 4/34 of patients with single biopsy from lingula or RML and 4/45 patients with 2 biopsies from other sites (p=0.6). There was 1 death in this series. CONCLUSIONS: Video-assisted thoracoscopic lung biopsy is an effective procedure. Single lung biopsy from lingula or RML compared to other anatomic sites has an equivalent diagnostic yield. PMID- 12627083 TI - Pneumonectomy vs lobectomy in the treatment of pathologic N1 NSCLC: could the type of surgical resection dictate survival? AB - AIM: To define the prognostic significance of specific types of N1 lymph node involvement in patients operated on for stage II (N1) NSCLC and to evaluate if the extent of resection affects survival. METHODS: Of 1117 patients operated on from 1985 to 1998, an homogeneous group of 124 consecutive patients with pathologic T1-T2 N1 disease who had undergone a complete resection with systematic nodal dissection were analysed. No patients received adjuvant radio- or chemotherapy. RESULTS: The overall 5-year survival rate was 48.8%. Survival was not related to pathologic T factor, histology, number, percentage or level of N1 involved, visceral pleura involvement, number of lymph nodes dissected. Patients were then divided into 3 groups depending on the level of lymph node involvement (stations 10, 11 and 12-13) and survival analysed according to the extent of resection (pneumonectomy vs lobectomy). No significant difference was found, however, in the group of level 10, patients treated by pneumonectomy showed a better 5-year survival (58%) compared to patients treated by lobectomy (33%) with a median survival of 110 against 58 months. This data was confirmed by a lower incidence of local recurrence in the pneumonectomy group than lobectomy group (0% vs 24%), whereas the same incidence of distant metastases was observed in the two groups (29% vs 23%). CONCLUSIONS: In patients with stage II (N1) NSCLC, only in case of station 10 involved, pneumonectomy could allow a better survival lowering the incidence of local recurrence. However the major part of patients with stage II (N1) NSCLC die for distant metastasis. This supports the necessity to develop a specific systemic treatment. PMID- 12627084 TI - The importance of the time interval between diagnosis and operation in myasthenia gravis patients. AB - AIM: Complete remission rates in patients with myasthenia gravis (MG) using anticholine esterases, immunosuppressive drugs, and medical therapy methods like plasmapheresis are low. Because high levels of complete remission and advantages are reported after thymectomy this surgical method is widely used in patients with MG. METHODS: The preoperative status and the responses to thymectomy have been studied in 15 patients who had been diagnosed as MG and underwent thymectomy in Ankara Numune Education and Research Hospital. RESULTS: Patients who underwent thymectomy have shown a high proportion of clinical improvement (73%). We could not establish a direct relationship between the results and the patients' age, sex features and the pathological characteristics of the excisional biopsy specimens. The benefit rates for patients who had a short period of time between diagnosis and operation (87.5%) were higher (91%). CONCLUSIONS: Thymectomy because of its high benefit rate is an advantageous therapeutic tool. The chance of benefiting from thymectomy increases when the history of MG is short and the stage of the disease is early. PMID- 12627085 TI - Lung transplantation in a Jehovah's witness. Case report in a twinning procedure. AB - Jehovah's Witnesses, patients who refuse blood transfusions, are generally not considered as candidates for lung transplantation owing to the frequent requirement for transfusions. A successful procedure in a Jehovah's Witness is presented and to our knowledge this is the 2(nd) reported case. The patient, a 38 year-old female, type I diabetes, affected by idiopatic pulmonary fibrosis underwent left lung transplantation. From the same pulmonary bloc a twinning procedure was obtained by means of right lung transplantation in a 58-year-old man affected by the same pathology. Surgical strategies employed in achieving a successful outcome, ethical and moral aspects are discussed. PMID- 12627086 TI - Life threatening complications caused by bronchogenic and oesophageal duplication cysts in a child. AB - An 11-year-old girl presented with severe respiratory distress, fever and septic manifestations. Computed tomography scan (CT) of the chest showed 2 separate superior and posterior mediastinal cysts, the upper one causing severe extrinsic compression of the trachea, and the oesophagus, while the lower cyst was at the subcarinal region compressing the 2 major bronchi. Emergency thoracotomy was performed permitting complete resection of intra-mural oesophageal enteric cyst, and a subcarinal bronchogenic cyst. The postoperative course was uneventful. PMID- 12627087 TI - Minimal invasive surgery for resection of parathyroid tumor in the aortopulmonary window. AB - Video-assisted thoracoscopic surgery for parathyroid glands deep in the mediastinum is a safe and successful procedure, but its success depends on accurate localization before operation. Recently, a variety of minimally invasive techniques to approach cardiovascular disease have been proposed to eliminate the morbidity of standard sternotomy. We describe a case involving a 71-year-old woman with persistent hyperparathyroidism resulting from an ectopic mediastinal parathyroid gland, which was removed through an upper sternotomy. The technique has proved safe, effective, and aesthetically acceptable to the patient. PMID- 12627088 TI - Pericardial retraction sutures facilitate intrapericardial pneumonectomy. PMID- 12627089 TI - Reducing the postoperative cost of carotid endarterectomy. Results of a randomized study. PMID- 12627090 TI - Malposition of a catheter in the persistent left superior vena cava. A rare complication of totally implantable venous devices. PMID- 12627091 TI - Treatment of femoral artery thrombosis following intra-aortic balloon insertion leaving the catheter in place. PMID- 12627092 TI - Esophago-broncho-cutaneous fistulae and tuberculous mediastinal lymphadenitis. PMID- 12627093 TI - Form expert panels to develop diagnosis and management protocols. PMID- 12627094 TI - Immunohistochemical detection of epidermal growth factor and its receptor in salivary gland carcinomas. PMID- 12627095 TI - The role of tongue reduction. AB - Macroglossia may occur as a congenital or acquired condition. The enlarged tongue has both functional and cosmetic deformity, which may affect the oral airway, speech, and the development of the jaws. We discuss the various tongue-reduction procedures and present cases to illustrate the uses of this procedure, its potential complications, and the results. PMID- 12627096 TI - Tongue piercing and its adverse effects. AB - Piercing has become so popular during the last 20 to 30 years that many physicians are now treating patients with piercings and dealing with its side effects. We present 3 cases that illustrate the complications of tongue piercing (ie, infection, bleeding, and embedded ornaments). We describe the methods for inserting the ornaments to illustrate the possible adverse effects. Treatment recommendations and their application to those 3 patients are described. PMID- 12627097 TI - Chronic inflammation around painless partially erupted third molars. AB - OBJECTIVES: We sought to assess the histologic host response in chronic, symptomless pericoronitis. STUDY DESIGN: Gingival mucosal (n = 20) and dental follicle (n = 20) samples were collected during extraction from patients with pericoronitis and clinically healthy control subjects. Antibodies-recognizing macrophages (CD68), natural killer cells (CD56), T cells (CD2), helper T cells (CD4), suppressor/cytotoxic T cells (CD8), and neutrophils (lactoferrin) were applied in a labelled streptavidin-biotin method by using a DAKO TechMate staining robot. RESULTS: Macrophage was the most numerous kind of cell in pericoronitis, but CD2+ T lymphocytes, with a normal CD4/CD8 ratio, were also increased (P < .01). Neutrophils were not increased and did not show signs of activation. Dental follicles did not contain increased numbers of inflammatory cells. CONCLUSION: This type of pericoronitis is a chronic/smoldering, rather than an acute/purulent, infection. Because of the chronic and often symptomless nature of pericoronitis, various long-term sequelae may result, which may lead to the need for extraction. PMID- 12627098 TI - Necrotizing fasciitis caused by dental infection: a retrospective analysis of 9 cases and a review of the literature. AB - OBJECTIVES: Necrotizing fasciitis of the head and neck is an uncommon, potentially fatal soft tissue infection characterized by extensive necrosis and gas formation in the subcutaneous tissue and fascia. The aims of this study were to describe the condition of this rare disease and to find factors affecting the mortality. STUDY DESIGN: Nine of our new cases and 125 reported cases in the English-language literature with necrotizing fasciitis of dental origin were reviewed. RESULTS: Two of our 9 patients had some form of systemic disease such as diabetes, cardiac insufficiency, renal failure, or cerebral infarction, whereas the other 7 had no particular general complications. A computed tomography examination was useful for detecting gas formation in the deep neck. All 9 patients underwent extensive debridement within 24 hours, and good results were obtained. In contrast, 24 of the 125 reviewed patients died despite therapy. Factors affecting the mortality were associated diseases such as diabetes or alcohol abuse, delay of surgery, and the complication mediastinitis. CONCLUSION: Necrotizing fasciitis is still a potentially fatal disease. Early and aggressive debridement may reduce mortality. PMID- 12627099 TI - Amalgam-contact hypersensitivity lesions and oral lichen planus. AB - OBJECTIVE: The purpose of this study was to investigate the relationship between amalgam restorations and oral lichen planus. STUDY DESIGN: Eighty-one patients with oral lichenoid lesions were characterized clinically and skin patch tested for amalgam or mercury hypersensitivity. Thirty-three of these patients had amalgam fillings in contact with oral lesions replaced and were followed to determine the outcome. RESULTS: Clinically, 2 patient groups were identified: (1) 30 patients with probable amalgam-contact hypersensitivity lesions (ACHLs) and (2) 51 patients with oral lichen planus (OLP) but no clear relationship with amalgam. Seventy percent of ACHL cases were patch test positive for amalgam or mercury compared with only 3.9% of OLP cases (P < .0001). Amalgam replacement resulted in lesion improvement in 93% of ACHL cases. No such improvement was observed in the OLP cases treated (P < .001). CONCLUSION: OLP is a heterogeneous condition within which an ACHL subgroup can be identified. ACHLs, but not other OLP lesions, respond favorably to amalgam replacement. A strong clinical association between lesions and amalgam restorations plus a positive patch test result was a good predictor of lesion improvement on amalgam replacement. PMID- 12627100 TI - Oral health and health behavior in patients referred for open-heart surgery. AB - OBJECTIVE: Little is known about the dental health and health behavior of patients referred for open-heart surgery because of severe heart disease. Yet, coronary atherosclerosis has been suggested to be an inflammatory disease in which chronic dental infections may trigger pathogenic mechanisms in the walls of arteries. In epidemiological studies periodontal disease in particular has been linked with coronary heart disease (CHD). STUDY DESIGN: We set out to investigate by means of clinical examination, x-ray, and questionnaire the oral health status, health-related behavior, and conceptions of 256 CHD patients referred for open-heart surgery in the Kuopio University Hospital. The patients' heart disease was New York Heart Association grades II-IV. For comparison, 250 age- and sex matched non-CHD patients were also studied. Differences between groups were analyzed conventionally and with the Total Dental Index (TDI), which uses linear modeling and logistic regression analyses to summarize periodontal factors and other signs of oral infections. We expected to find neglected dental care and worse oral health situation among the CHD patients. RESULTS: Edentulousness was seen in 35% of the CHD group and in 15% of the non-CHD group (P < .001). The CHD group had significantly lower number of teeth (8.8 +/- 9.1) than the non-CHD group (17 +/- 10.5; P < .001). No dental appointment in the past 10 years was reported in 66% of the CHD patients and 35% of the non-CHD patients. Forty-nine percent of the CHD group and 69% of the non-CHD group had had a dental visit within the past 2 years (P < .001). Daily tooth brushing was reported by 46% of the CHD group and 56% of the non-CHD group (P < .05). Only 2% of the CHD group and 8% of the non-CHD group flossed daily (P < .001). TDI scores were significantly higher in the edentulous than in the dentate study groups (P < .001), but no difference was observed between CHD and non-CHD patients of the same dentate or edentulous status. Mean TDI score was 3.2 +/- 0.8 in edentulous CHD patients and 3.1 +/- 0.4 in edentulous non-CHD patients; the scores in the dentate patients were 2.5 +/- 0.9 in the CHD group and 2.4 +/- 2.0 in the non-CHD group. The TDI score appeared almost significant a predictor of CHD when used as a dependent variable in logistic regression analysis (P = .08). CONCLUSION: Worse oral health status and less satisfactory oral health behavior were observed in the CHD patients than among the cardiologically healthy. PMID- 12627101 TI - Budesonide: a novel treatment for oral chronic graft versus host disease. AB - OBJECTIVE: This clinical trial aims to evaluate the efficacy of budesonide, a newly registered steroid with high potency and low bioavailability, for the treatment of chronic oral graft versus host disease (GVHD). STUDY DESIGN: Twelve patients with chronic resistant oral GVHD were treated with 3 mg budesonide/5 ml saline 2 to 3 times a day for up to 3 months. Oral manifestations were monitored, and mucosal response scored. RESULTS: All patients responded positively to the mouthwash, and 7 of the 12 patients were scored as having "good" or "complete" recovery by both examiner and subject. An early response noted within the first 2 to 3 weeks of treatment was complemented by a probable cumulative effect seen during the first months of treatment. CONCLUSION: Budesonide is suggested as an alternative treatment for chronic oral GVHD. PMID- 12627102 TI - Cheilitis glandularis: an unusual histopathologic presentation. AB - Cheilitis glandularis (CG) is an uncommon disease that usually affects the lower lip of adults. It is characterized by enlargement and eversion of the lip in association with excretory duct dilatation. The presence of minor salivary gland hyperplasia is controversial. Three types of CG have been described in the literature; the classification is based on the common clinical and histopathologic findings (ie, simple, superficial, and deep). This report is of an unusual case of CG simplex for which the initial histopathologic diagnosis was papillary cystadenoma, a neoplastic process. The lesion was completely excised, and final microscopic review of a larger specimen revealed chronic sclerosing sialadenitis, dilated salivary secretory ducts with oncocytic change and periductal inflammation, and foci of adenomatous hyperplasia consisting of enlarged ducts exhibiting squamous epithelial metaplasia and hyperplasia with papillary architecture. The latter proliferative pattern is an unusual finding in what is otherwise clinically diagnosed as CG. PMID- 12627103 TI - Pagetoid reticulosis: a case report and review of the literature. AB - The histologic finding of a pagetoid epidermotropic growth pattern is associated with a variety of conditions that range from reactive to neoplastic in nature. Included among these conditions is the uncommon skin disorder pagetoid reticulosis. Pagetoid reticulosis may present clinically as a solitary, indolent plaque-like lesion. Conversely, it may manifest as a more generalized clinically aggressive disease. The generalized form may develop into systemic lymphoma, leading to severe morbidity and death. We present a case of pagetoid reticulosis of the perioral region, a rare anatomic site for this condition. Histopathologic and immunohistochemical findings were consistent with a diagnosis of isolated pagetoid reticulosis. This review discusses the nature of pagetoid reticulosis and addresses past and current concepts regarding the condition. PMID- 12627104 TI - Sweet's syndrome: recurrent oral ulceration, pyrexia, thrombophlebitis, and cutaneous lesions. AB - We report a case of Sweet's syndrome with recurrent oral ulceration, pyrexia, skin lesions, and migratory thrombophlebitis, with no detectable systemic cause, during a 2-year follow-up. Biopsy examination both of oral lesions and the skin eruption showed a characteristic dense, perivascular, neutrophilic infiltrate in the lamina propria. Laboratory investigations confirmed an inflammatory syndrome with an increased erythrocyte sedimentation rate, but no underlying cause was found. Sweet's syndrome is a rare immunologically mediated condition that belongs to the group of neutrophilic dermatoses that must be differentiated particularly from Behcet's disease. It is characterized by red-brown plaques and nodules that are frequently painful and occur primarily on the head, neck, and upper extremities. Often the patients also have neutrophilia and fever and may have oral ulceration. In approximately 10% of patients with Sweet's syndrome, there is an associated malignancy--most commonly acute myelogenous leukemia--but some cases, as here, are unassociated with detectable malignant or other disease, although the syndrome may precede the onset of definable systemic disease. PMID- 12627105 TI - Synchronous mucoepidermoid carcinoma of tongue and pleomorphic adenoma of submandibular gland. AB - Presentation with synchronous salivary gland tumors is rare, with the most typical combination being Warthin's tumor and pleomorphic adenoma of the parotid gland. Involvement of minor salivary glands in such occurrences is extremely uncommon. We report a case of simultaneous mucoepidermoid carcinoma of the tongue and pleomorphic adenoma of the submandibular gland in a 40-year-old woman. The submandibular mass was initially considered to represent regional lymph node tumor metastasis but later was shown to be an intraglandular primary neoplasm. PMID- 12627106 TI - Clear-cell variant of calcifying epithelial odontogenic tumor: clinical and radiographic characteristics. AB - OBJECTIVES: Clear cells have been reported in approximately 8% of cases of calcifying epithelial odontogenic tumor (CEOT). The purpose of this study was to describe the clinical and radiographic features of clear-cell CEOT (CCEOT). STUDY DESIGN: Eighteen cases of CCEOT were identified by review of the literature from 1958 to 2001, and a new one was added (total 19; 12 central, 7 peripheral). RESULTS: Fourteen tumors (74%) were located in the mandible, with the central lesions favoring the posterior area and the peripheral lesions the anterior bicuspid area. Radiographic features were as follows: 50% radiolucent, 50% mixed radiolucent-radiopaque; 92% unilocular; 64% well-defined noncorticated borders, 27% well-defined corticated borders, 9% irregular borders. Cortical perforation was common (67%) compared with CEOT without a clear-cell component (6.7%). Recurrence was reported in 17% of the central lesions and none of the peripheral ones. CONCLUSIONS: CCEOT is a distinct variant of CEOT; its high tendency for cortical perforation may indicate a more aggressive behavior. PMID- 12627107 TI - Estrogen receptors in human pulp tissue. AB - OBJECTIVE: The aim of this study was to evaluate the expression of estrogen receptor (ER)-alpha in human pulp tissue. STUDY DESIGN: Tissue samples were collected from 51 teeth extracted for orthodontic reasons or from endodontically treated teeth. Immunohistochemical staining was performed with mouse antihuman ER alpha 1D5 by using the alkaline phosphatase antialkaline phosphatase technique. Immunohistochemical reactivity was semiquantitatively assessed and graded on an intensity scale of 0 to 3. RESULTS: ERs were found in odontoblasts, endothelial cells, and Schwann cells. By means of statistical analysis, ERs were found to be common in odontoblasts (1.3-1.6) and in endothelial cells (1.0-1.4), whereas in Schwann cells they were somewhat rare (0.7-1.1). Difference in expression of ERs was not found among patients in different age groups. With respect to sex, a difference in reactivity was observed in Schwann cells, which showed higher reactivity in female pulps. CONCLUSION: The results of this study confirm the expression of ER-alpha in the human dental pulp. PMID- 12627108 TI - pH and calcium ion release of 2 root-end filling materials. AB - OBJECTIVE: The purpose of this study was to evaluate the pH and calcium ion release of 2 materials used for root-end fillings and perforation repair. STUDY DESIGN: ProRoot and MTA-Angelus were placed in plastic tubes and immersed in glass flasks containing deionized water. After 3, 24, 72, and 168 hours, the water in which each had been immersed was tested to determine the pH changes and released calcium. RESULTS: The values for pH and calcium ion release were slightly higher for MTA-Angelus than ProRoot. The calcium release and pH of both materials were initially higher. CONCLUSION: The results suggest that both materials release calcium and promote alkaline pH. PMID- 12627109 TI - Calcium hydroxide reduces lipopolysaccharide-stimulated osteoclast formation. AB - OBJECTIVE: The purpose of this study was to determine the direct effects of lipopolysaccharide (LPS) on osteoclastogenesis and to assess the ability of calcium hydroxide-Ca(OH)(2)-to inhibit the osteoclast formation stimulated by LPS. STUDY DESIGN: RAW 264.7 cells were cultured with 50 ng/mL recombinant receptor activator of NF-kappaB ligand (RANKL) for 72 hours. RANKL was then removed, and the cells were treated with 0, 1, 10, or 100 ng/mL of LPS, Ca(OH)(2) treated LPS, or 50 ng/mL of RANKL as a positive control for an additional 48 hours. Cells were fixed and stained with fluorescein isothiocyanate-conjugated phalloidin to detect actin ring formation, and histochemistry was performed to detect multinucleated cells expressing tartrate-resistant acid phosphatase activity. RESULTS: LPS induced osteoclast-like cell (OCL) formation in a dose dependent manner when osteoclast precursor RAW 264.7 cells were pretreated for 72 hours with RANKL. Ca(OH)(2) significantly inhibited the ability of LPS to stimulate OCL formation. CONCLUSION: This study shows that LPS directly stimulates OCL formation. The detoxification of LPS by treatment with Ca(OH)(2) significantly reduced its ability to trigger the differentiation of OCLs. PMID- 12627110 TI - Extraoral endodontic treatment by retrograde insertion of posts: a long-term study on replanted and transplanted teeth. AB - OBJECTIVE: The aim of the study was to investigate the healing results of teeth replanted or transplanted in different indications and treated by extraoral root canal therapy. STUDY DESIGN: Extraoral root canal treatment was performed from a retrograde direction with posts made of ceramics or titanium. Preoperatively, the pulp status was classified as definitely infected (n = 47) or not infected/symptomless (n = 78), and the condition of the periodontal ligament (PDL) was classified as damaged (n = 50) or not damaged (n = 75) according to trauma type and extraoral storage. The healing after replantation or transplantation was diagnosed as functional, ankylosis, or infection depending on clinical and radiographic findings. RESULTS: In total, 125 teeth in 99 patients were replanted or transplanted. Mean observation period was 44.4 months. Teeth with a less damaged PDL in no case exhibited ankylosis/replacement resorption after a mean observation period of 53 months. Teeth without preoperative infection of the pulp in no case showed infection-related complications after a mean observation period of 30 months. Teeth classified as definitely infected before treatment had a high frequency of continuous periradicular bone and root resorption. CONCLUSIONS: The extraoral insertion of posts appeared to inflict no additional damage to the PDL that was clinically relevant. Infection-related complications were prevented. Teeth that were classified as definitely infected at the time of treatment had a low rate of healing and should undergo conventional root canal disinfection before this surgical procedure is applied. PMID- 12627111 TI - Trabecular alveolar bone in the human mandible: a dual-energy x-ray absorptiometry study. AB - OBJECTIVE: The purpose of this study was to evaluate the potential use of dual energy x-ray absorptiometry for the assessment of bone mineral content and bone mineral density before implant placement. MATERIALS AND METHODS: The material examined consisted of 63 mandibular bone specimens cut from 21 fresh cadavers (11 men; 10 women). Three specimens were cut per cadaver in the incisal, premolar, and molar regions. Three regions of interest (G, R1, and R2) were delineated. The global bone specimen (G) consisted of the whole specimen (ie, both cortical and trabecular bone). R1 and R2 were delineated in the trabecular bone of the alveolar process. RESULTS: In all subjects, the combined bone mineral content of the whole mandible specimens (global bone mineral content) was significantly correlated with age. The difference between the mean bone mineral densities of the male and female mandibles was found to be significant for G (P = .009). The mean bone mineral densities of dentate and edentulous specimens were also found to be significantly different for G and R1, respectively (P = .001 and P = .02), but not for R2. A positive correlation could be detected among the mean bone mineral density of G and R1, G and R2, and R1 and R2 of (1) male and female specimens, (2) dentate and edentulous specimens, and (3) incisal, premolar, and molar specimens. CONCLUSIONS: The intra-alveolar trabecular bone of these 21 mandibles is affected by the same local and systemic influences as cortical bone, whereas the infra-alveolar trabecular bone is mostly sensitive to dental status. The cortical and trabecular bone of the 10 mandibles from women is more sensitive to systemic influences, whereas that from men is more sensitive to local influences. This is somewhat in agreement with some studies that found an association between osteoporosis and oral bone loss, which is a metric measure. PMID- 12627132 TI - Late bullet migration into the knee joint. AB - A 25-year-old active-duty police officer was found to have an intra-articular foreign body on radiographic study of his left knee joint. He had a gunshot wound to the midthigh 54 months prior to the presentation of symptoms. The bullet was lodged in the soft tissue without involving neurovascular structures. The patient complained of limited range of motion of the joint and a "rattle" sensation of the knee. Arthroscopically, a deformed metallic foreign body was found and retrieved. There was no injury inside the joint related to the loose body. These findings were consistent with a migrating bullet from the midthigh to the knee joint. The patient recovered uneventfully and returned to work. PMID- 12627112 TI - A comparison of a new limited cone beam computed tomography machine for dental use with a multidetector row helical CT machine. AB - OBJECTIVE: We sought to compare a new limited cone beam computed tomography (CT) machine for dental use (3DX) with the multidetector CT machine for image quality and skin doses. STUDY DESIGN: Images of the right maxillary central incisor and the left mandibular first molar of an anthropomorphic phantom were taken by both the 3DX and the multidetector CT. A 5-point method was used to evaluate the depiction of cortical and cancellous bone, enamel, dentin, pulp cavity, periodontal ligament space, lamina dura, and overall impressions. Furthermore, the skin doses for both modalities were compared. RESULTS: The image quality of the 3DX was better than the multidetector CT for all items (P < .01). Moreover, the mean skin doses with the multidetector CT were 458 mSv per examination, whereas the doses with the 3DX were 1.19 mSv per examination. CONCLUSIONS: These results clearly indicate the superiority of the 3DX in the display of hard tissues in the dental area while substantially decreasing the dose to the patient. PMID- 12627133 TI - Unusual solitary ganglion cysts of the anterior segment of the lateral meniscus. AB - Generally, ganglion or synovial cysts have been reported to originate from numerous structures in and around the knee joint, including the anterior cruciate ligament, posterior cruciate ligament, popliteal tendon, and menisci. Nevertheless, a ganglion is not a synovial cyst and there is no synovial lining. Meniscal cysts present a subgroup of ganglion cysts that occur in association with meniscal tears. We report 2 cases of unusual meniscal cyst from the anterior segment of the lateral meniscus without any meniscal tear and discuss the possibility of their pathogenesis and treatment. PMID- 12627134 TI - Intra-articular hemangioma of the knee. AB - Intra-articular hemangioma of the knee is a rare cause of pain and spontaneous hemarthrosis, often seen as an internal derangement of the joint in children and young adults. The tumor is observed in two different forms: the synovial hemangioma or the arteriovenous malformation named also as hemangiohamartomas. They may cause hemorrhagic synovitis and arthropathy, probably as a result of recurrent episodes of intra-articular bleeding and mechanical irritation. Four cases of intra-articular hemangioma of the knee are presented in this study. Two of our cases were diagnosed as synovial hemangioma with the lesions localized inside the knee and the other 2 were intermediate type with the tumor extending to the muscle group and the skin. After magnetic resonance imaging and angiographic assessment, arthroscopic excision was performed in the localized type of the tumor. The other 2 cases received only diagnostic arthroscopy and biopsy followed by conservative treatment due to the extensive localization of the tumors and the serious chondral lesions. The decision of the treatment modality is very difficult in intra-articular hemangioma of the knee because, when combined with chondral degeneration, the incidence of local recurrence is high. PMID- 12627135 TI - Avulsion fracture of the posterior oblique ligament associated with acute tear of the medial collateral ligament. AB - The term posterior oblique ligament was proposed in 1973 but the specific anatomy of this region has been controversial. It has been concluded that the posterior oblique ligament is separated from the medial collateral ligament, each with distinctly different bony attachment points. In this article, we report a posterior oblique ligament avulsion fracture associated with a medial collateral ligament rupture. PMID- 12627136 TI - Intra-articular migration of femoral interference screw: Open or arthroscopic removal. AB - Migration of the femoral interference screw in to the knee joint following anterior cruciate reconstruction is a rare complication. A migrated interference screw usually requires removal as it often results in mechanical symptoms. Formal arthrotomy may be required to remove a dislodged screw lying in the femoral notch because arthroscopic removal can be difficult or not possible in the presence of an intact integrated anterior cruciate ligament graft or scarring in the notch. When attempting arthroscopic removal in a similar situation, one should foresee the possibility of a formal arthrotomy and this should be discussed with the patient and consent obtained preoperatively. PMID- 12627137 TI - A reusable suture anchor for arthroscopy psychomotor skills training. AB - For residents to adequately develop the early arthroscopy psychomotor skills required to better learn how to manage the improvisational situations they will encounter during actual patient cases, they need to experience sufficient practice repetitions within a contextually relevant environment. Unfortunately, the cost of suture anchors can be a practice repetition-limiting factor in learning arthroscopic knot-tying techniques. We describe a technique for creating inexpensive reusable suture anchors and provide an example of their application to repair the anterior glenoid labrum during an arthroscopy psychomotor skills laboratory training session. PMID- 12627139 TI - Arthroscopic anterior cruciate ligament reconstruction: An air-fluid medium to enhance visual clarity. AB - We describe a surgical technique that has been used successfully during arthroscopic anterior cruciate ligament reconstruction to enhance the visual clarity of the operating field specifically during tunnel placement. The precise siting of both tibial and femoral tunnels is critical to both the short- and long term success of this procedure. Gentle insufflation of the knee joint with air prior to tunnel siting allows for an excellent view of the intercondylar notch. This visual clarity helps in the precise placement and measurement of the femoral tunnel. PMID- 12627138 TI - Snapping knee caused by intra-articular tumors. AB - Locking of the knee can present with no history of antecedent injury. We identified impingement of intra-articular tumors in 2 cases. Intra-articular tumors are relatively rare. Mechanical symptoms were present in both cases. On physical examination, there was a palpable mass at the medial region of the patellofemoral joint. The interior had been replaced mainly by amorphous necrotic tissue. The definite diagnosis of a soft-tissue mass of the knee could not be made on histologic examination. PMID- 12627140 TI - Arthroscopic surgery in the posterior compartment of the knee: Suture fixation of anterior and posterior cruciate ligament avulsions. AB - Anterior and posterior cruciate ligament avulsion injuries in the posterior compartment of the knee are most commonly dealt with using open approaches. We report an arthroscopic method of treatment using sutures, without the need of any metal implants, that can be safely used in patients with open physes and without the need of a 70 degrees arthroscope. PMID- 12627141 TI - Vascular leiomyoma presenting as medial joint line pain of the knee. AB - We present the case of a 27-year-old woman with a subcutaneous vascular leiomyoma occurring around the knee. The patient presented with a several-year history of medial-sided knee pain with an unknown etiology. The clinical presentation, magnetic resonance imaging, and histopathologic features are reported, and the literature pertaining to the clinicopathologic characteristics is reviewed. By including vascular leiomyomas in the differential of painful lower extremity subcutaneous masses, an early accurate diagnosis is possible. PMID- 12627142 TI - Arthroscopic removal of a loose body osteophyte fragment after superior patellar dislocation with locked osteophytes. AB - The authors report the case of a loose body from a fractured osteophyte after a superiorly dislocated patella with locked osteophytes. Few cases of superiorly dislocated patellae have been reported in the literature and no cases of osteophyte fracture fragments after locked osteophytes with subsequent arthroscopic loose body removal have been reported. The loose body was removed and the distal pole of the patella was debrided arthroscopically. This patient and the majority of previously reported cases, herein reviewed, had patella alta with pre-existing patellofemoral arthrosis. Patella alta in the face of patellofemoral arthrosis should be considered a risk factor for loose body formation. Therefore, recurrent superior patellar dislocation and locking osteophytes may be a relative indication for pre-emptive arthroscopic debridement of locked osteophytes. PMID- 12627143 TI - Septic arthritis caused by Erysipelothrix rhusiopathiae infection after arthroscopically assisted anterior cruciate ligament reconstruction. AB - A case of septic arthritis caused by Erysipelothrix rhusiopathiae, after an arthroscopically assisted anterior cruciate ligament (ACL) substitution in a non immunosuppressed patient is described. An 18-year-old man underwent an ACL reconstruction with a quadruple hamstring graft. Eight days postoperatively, the patient developed fever, knee pain, and effusion without erythema or suppuration. He was readmitted to the hospital with the diagnosis of septic arthritis. The patient's erythrocyte sedimentation rate, C-reactive protein level, and white blood cell count were high. The joint was aspirated and the fluid was sent for cultures that revealed the presence of E rhusiopathiae. E rhusiopathiae is widespread in nature, it is transmitted by direct cutaneous laceration, and it causes septic arthritis, meningitis, endocarditis, and renal failure in immunosuppressed people with poor prognosis. In our case, the infection was treated with arthroscopic lavage and debridement, retention of the graft and hardware, and intravenous antibiotic administration for 6 weeks, followed by oral administration for 16 weeks. PMID- 12627144 TI - Torsion of an intra-articular lipoma as a cause of pseudo locking of the knee. AB - We describe a case of a painful locked knee caused by the torsion of an intra articular lipoma and review the literature on this unusual condition. There are many cases of locked knee and of intra-articular lipoma but to our knowledge there are no previous documented cases of a torted lipoma being identified as a cause of locking. PMID- 12627145 TI - Arthroscopic capsular shrinkage of the shoulder for the treatment of patients with multidirectional instability: Minimum 2-year follow-up. AB - PURPOSE: The study goal was to evaluate the arthroscopic treatment of patients with multidirectional instability of the shoulder (MDI). MDI is a disabling condition as a result of pain and restriction of activity. The treatment of MDI is primarily nonsurgical (supervised physiotherapy). TYPE OF STUDY: Prospective cohort study. METHODS: Arthroscopic capsular shrinkage, using a bipolar radiofrequency system, was performed in 32 patients (33 shoulders). The mean age was 27 years (range, 15 to 49). The Constant score was used for assessment preoperatively and at 6-month intervals. In 8 patients, labral lesions were revealed at arthroscopy and were addressed using arthroscopic methods. RESULTS: The follow-up was from 24 to 33 months (26 mean). Patients without labral pathology (group A) and patients with labral lesions (group B) are discussed separately. Group A showed an increase in the mean Constant score at 6 months, but the scores reached a plateau after 6 months. Statistical analysis using 1-way analysis of variance (ANOVA) showed that the mean difference between the preoperative Constant score and the score at latest follow-up was statistically significant (P <.0001; 95% confidence interval, 13.4 to 31.6; standard error, 2.2). The rate of failure or complication was 16%. Three patients experienced recurrence of instability and another developed adhesive capsulitis. Overall, patient satisfaction was 83%. Group B showed an improvement in the Constant scores, and all 8 patients were satisfied. No clinical recurrence of the instability was seen in this group. However, 1 patient developed adhesive capsulitis. This group is too small to draw any statistically valid conclusions. CONCLUSIONS: The early results are encouraging. Postoperative physiotherapy is of paramount importance. Continued long-term follow-up of this relatively new technique is required to determine the failure rate increase that is likely to occur at longer time intervals. PMID- 12627146 TI - All-arthroscopic versus mini-open rotator cuff repair: A long-term retrospective outcome comparison. AB - PURPOSE: The goal of the study was to compare the results of all-arthroscopic rotator cuff repair with those of our mini-open rotator cuff repair. TYPE OF STUDY: Retrospective outcome study. METHODS: Sixty-four shoulders (58 patients) were identified; 35 in the all-arthroscopic group and 29 in the mini-open group. Average follow-up for all patients was 44.6 months, with a minimum of 24 months. The all-arthroscopic group included 3 small tears (< 1 cm), 24 medium-size tears (1 to 3 cm), and 9 large tears (3 to 5 cm). The mini-open group included 2 small tears, 9 medium tears, and 18 large tears. All patients in both groups underwent arthroscopic assessment with arthroscopic subacromial decompression. None underwent formal acromioclavicular joint resection. Four of the all-arthroscopic and 11 of the mini-open patients underwent coplaning of the acromioclavicular joint. Anchors were the primary method of arthroscopic fixation, with an average of 1.5 anchors per case. RESULTS: The average final follow-up UCLA score for the arthroscopic group was 32.6 and for the mini-open group was 31.4, and the average final follow-up ASES score for the arthroscopic group was 91.7 and for the mini open group was 90.0. No patients in the arthroscopic group developed fibrous ankylosis, whereas 4 patients in the mini-open group developed the condition (14%). No anchor-related complications were noted. Shoulders in the all arthroscopic group showed greater motion at 6 and 12 weeks postoperatively and slightly better motion at final review. However, final motion difference was not statistically significant. Overall, 4 patients (6.3%) demonstrated some tenderness in the acromioclavicular joint on palpation, but none had clinical symptoms requiring treatment. CONCLUSIONS: All-arthroscopic cuff repair provides comparable outcomes and complication rates to arthroscopic decompression with mini-open repair. The lower incidence of fibrous ankylosis favors the all arthroscopic technique. A trend for better early motion was also noted in the all arthroscopic group. Analysis of variance shows no measurable outcome difference between tear sizes. PMID- 12627147 TI - Rotator cuff repair with bioabsorbable screws: An in vivo and ex vivo investigation. AB - PURPOSE: The purpose of this study was to evaluate in vivo the clinical outcomes of rotator cuff repairs with bioabsorbable screws compared with metal suture anchors, and to compare the ex vivo initial load to failure of rotator cuff repairs using 3 different bioabsorbable screws, suture anchors, and transosseous sutures. TYPE OF STUDY: In vivo clinical outcomes investigation, and ex vivo biomechanical study. METHODS: Three cohorts of patients with rotator cuff tears that measured less than 4 cm(2), were sequentially repaired with Mitek Rotator Cuff QuickAnchors (Mitek Surgical Products, Norwood, MA) (n = 9), Arthrex Headed Bio-Corkscrews (n = 9) (Arthrex, Naples, FL), and Mitek Rotator Cuff QuickAnchors (n = 9). Patients were systematically assessed with a specific shoulder questionnaire and 23 shoulder tests performed preoperatively and at 1 and 6 weeks, 3 and 6 months, and 1 year postoperatively. A correlative ex vivo biomechanical study was performed on 53 ovine shoulders to evaluate the initial failure load properties of bioabsorbable screws compared with fixation with suture anchors and transosseous sutures. RESULTS: In the in vivo portion of the study, the cohort treated with the Headed Bio-Corkscrew demonstrated no improvement on any measured parameter until 1-year after rotator cuff repair. In contrast, shoulders repaired with Mitek Rotator Cuff QuickAnchors demonstrated improved overall shoulder function as early as 6 weeks postoperatively (P =.002), had a better constant score at 1-year after repair (88 +/- 9 v 73 +/- 17; P =.016), and a lower rate of revision rotator cuff repair (P =.029). In the ex vivo portion of the study, the bioabsorbable headed screws, Headed Bio-Corkscrew (100 +/- 30 N) and BioTwist (76 +/- 35 N), had inferior initial failure load properties compared with suture anchors (140 +/- 36 N) and transosseous sutures (147 +/- 68 N). In contrast, the BioCuff (190 +/- 56 N), a bioabsorbable implant that used a screw and serrated washer design, had equivalent initial failure load properties as the suture repairs. CONCLUSIONS: This investigation had poorer early outcomes, a lower shoulder functional score 1-year after repair, and a higher rate of repeat surgery in patients who had their rotator cuff repaired with a bioabsorbable screw than in patients who had their shoulders repaired with a standard metal suture anchor. Furthermore, the biomechanical testing demonstrated a lower tensile load to failure in the tendons repaired with a simple screw design compared to suture anchors with a mattress stitch. Of note, the implant that used a screw and washer design demonstrated a greater ability to resist initial tensile load. PMID- 12627148 TI - Arthroscopic repair of full-thickness supraspinatus tears (small-to-medium): A prospective study with 2- to 4-year follow-up. AB - PURPOSE: The objective of this study is to report on the complete arthroscopic repair of full-thickness tears of the supraspinatus. TYPE OF STUDY: Prospective cohort study. METHODS: Between 1995 and 1999, 139 full arthroscopic rotator cuff repairs were performed; 37 were repairs of full-thickness supraspinatus tears. Between 1997 and 1999, there were 24 patients who had a complete arthroscopic repair of supraspinatus tears. The preoperative and postoperative status of these patients was analyzed using the Constant score, American Shoulder and Elbow Society Index (ASES Index), a Visual Analog Pain Scale (VAS), a single question of percent function compared with the opposite unaffected extremity, and a single question reflecting satisfaction, "Would you undergo the surgery and the postoperative rehabilitation to achieve the result you have today?" There were 37 patients who had preoperative to postoperative VAS, percent function, postoperative acromiohumeral interval, and clinical rerupture incidences evaluated. The null hypothesis was tested and there was no difference between the preoperative to postoperative outcomes. Two groups were evaluated, those who had concomitant decompression and those who did not. RESULTS: The null hypothesis was not supported. The 4 scoring systems used for evaluation showed statistically significant improvement from preoperative to postoperative. There were no differences in outcome based upon sex or age as a variable. All patients would have surgery again to achieve their postoperative state. CONCLUSIONS: The arthroscopic repair of supraspinatus tears is effective for improving the functional status of the shoulder. PMID- 12627149 TI - Comparison of sagittal obliquity of the reconstructed anterior cruciate ligament with native anterior cruciate ligament using magnetic resonance imaging. AB - PURPOSE: This study was conducted to compare the obliquity of asymptomatic anterior cruciate ligament (ACL) grafts with normal controls using sagittal magnetic resonance imaging (MRI). TYPE OF STUDY: Case control study. METHODS: Sagittal MRIs from 30 patients with a reconstructed ACL graft and from 30 individuals with an intact ACL were reviewed. Reconstructed patients were operated on with a 2-incision technique using a patellar tendon autograft. These selected patients had a normal or nearly normal IKDC score with a 3 mm or less anterior posterior translation on KT-1000 arthrometer testing compared with the intact knee. MRI showed a continuous and homogeneous graft without evidence of roof impingement. Obliquity of the grafted ACL was determined on each lateral MRI by measuring the intersection of the graft line with the tibial plateau plane. These figures were compared with data similarly obtained from 30 individuals with a stable knee and an intact ACL determined by history and physical examination. RESULTS: Graft obliquity in reconstructed patients averaged 67 degrees with a range between 55 degrees and 81 degrees. In normal controls, intact ACL obliquity averaged 51 degrees with a range between 45 degrees and 55 degrees. The difference between the two groups was statistically significant (P <.0001). CONCLUSIONS: MRIs of patients with an appropriate tibial tunnel placement in order to avoid notch impingement showed a continuous and homogeneous graft similar to the native ACL, but with a more vertical graft that does not recreate the normal sagittal obliquity. However, according to arthrometer testing, these more vertical grafts can control anterior posterior knee displacement. PMID- 12627150 TI - Arthroscopic evaluation of articular cartilage lesions in posterior-cruciate ligament-deficient knees. AB - PURPOSE: The goal of this study was to gain more information on the likelihood of developing cartilage lesions in posterior cruciate ligament (PCL)-deficient knees. TYPE OF STUDY: Retrospective clinical study. METHODS: Standardized arthroscopy records of 181 patients with a nonsurgically treated acute or chronic PCL injury were analyzed with respect to cartilage degeneration. Subgroups with different duration of PCL insufficiency, the influence of isolated PCL or combined PCL/posterolateral instability, and the grade of posterior laxity was analyzed. RESULTS: PCL insufficiency significantly increased the risk of developing medial femoral condyle and patellar cartilage degeneration over time. Of patients whose PCL deficiency was present for more than 5 years, 77.8% showed degenerative cartilage lesions of the medial femoral condyle and 46.7% showed cartilage degeneration of the patella. After 1 year of PCL insufficiency, the number of medial femoral cartilage lesions increased threefold (13.6% v 39.1%). With the presence of combined PCL/posterolateral insufficiency the amount of medial femoral degeneration was significantly increased (36.6% v 60.6%). CONCLUSIONS: We found that PCL insufficiency is not a benign injury with respect to the development of degenerative articular cartilage lesions. The early and continuous increase in cartilage degeneration at the medial femoral condyle and the patella should be considered when discussing operative versus conservative treatment for a PCL-deficient knee. The rapid development of medial arthritis should also be considered during decision making, particularly in patients with combined PCL/posterolateral instability or those who underwent previous partial medial menisectomy. PMID- 12627151 TI - Usefulness of arthroscopic surgery in hip trauma cases. AB - PURPOSE: The goal of this study is to clarify the usefulness of arthroscopic surgery as an adjunct in the management of hip trauma. TYPE OF STUDY: Clinical case series. METHODS: Eleven joints from 10 hip trauma cases were studied. All joints had been injured in traffic accidents. Seven joints were classified as Thompson and Epstein (T-E) type I, 2 joints as type II, and 1 joint as type IV. The remaining case was a fracture of the acetabular weight-bearing region that was not included within these classifications. The 5 femoral head fractures associated with hip dislocation were further classified according to Pipkin: 3 joints were type 1, 1 joint was type 2, and 1 joint was type 3. The interval from injury to arthroscopy ranged from 1 to 7 days, with a mean of 3.2 days. RESULTS: In 7 cases, small free osteochondral or chondral fragments that were not detectable on either plain radiographs or computed tomography scans were seen arthroscopically. These fragments were debrided. Arthroscopic osteosynthesis using absorbable pins and fragment extraction were performed in 1 case each of the Pipkin's type 1 femoral head fractures. In the case of the acetabular weight bearing region fracture, reduction and percutaneous pinning were performed under arthroscopic observation. After a mean postoperative follow-up period of 9 years and 6 months, no abnormalities were observed in 9 of the joints. However, the T-E type IV joint developed osteoarthritis and the Pipkin type 3 joint developed aseptic osteonecrosis of the femoral head. CONCLUSIONS: Arthroscopic surgery allows procedures such as washout and debridement of small free osteochondral and chondral fragments, which were previously neglected in T-E type I and II dislocations despite the fact that they may cause osteoarthritis. In addition, arthroscopic surgery allows procedures such as the extraction or osteosynthesis of fracture fragments in cases associated with femoral head fractures. PMID- 12627152 TI - The effect of suture anchor design and orientation on suture abrasion: An in vitro study. AB - PURPOSE: To evaluate the effects of suture anchor design and orientation on suture abrasion in a cyclic model. TYPE OF STUDY: In vitro. METHODS: Biomechanical studies have shown suture breakage to be a predominant mode of failure in a suture anchor repair construct. It is possible that suture abrasion during knot tying or in vivo cyclic loading may contribute to early failure. This study specifically investigates suture abrasion caused by 17 commonly used suture anchors and demonstrates the effects of suture anchor angulation and rotation on suture abrasion. To eliminate target tissue as a source of failure, all anchors were implanted into a solid block of sawbones material and tested with No. 2 Ethibond Excel sutures (Ethicon, Somerville, NJ). The testing model focused on 3 variables: suture anchor type, suture pull angle (SA) and angle of anchor rotation (RA). Abrasion testing was then performed on a servohydraulic materials testing system by continually cycling the suture back and forth through each anchor with an excursion of 4 cm at a rate of 0.5 Hz under a load of 10 N until suture failure occurred. RESULTS: Sutures performed significantly better when cycled in line with the anchor at 0 degrees SA with 0 degrees RA than they did at 45 degrees SA with 0 degrees RA or 45 degrees SA with 90 degrees RA. We found no significant difference between anchors tested at 45 degrees SA with 0 degrees RA and 45 degrees SA with 90 degrees RA. For tests performed using metallic suture anchors, all constructs failed by fraying of the suture. Constructs using biopolymer anchors and nonabsorbable polymeric anchors experienced a mixture of suture and anchor eyelet failures. CONCLUSIONS: In addition to the statistically significant detrimental effects of suture anchor angulation and rotation on suture abrasion, suture anchor eyelet design may also influence suture abrasion. Surgeons should be aware of the effects of anchor angulation, suture position in the eyelet, and design and composition of the eyelet to maximize the durability of the construct. PMID- 12627153 TI - Accuracy and reliability of arthroscopic estimates of cartilage lesion size in a plastic knee simulation model. AB - PURPOSE: The goal of the study was to determine the accuracy and reliability of arthroscopic percent area estimates in a plastic knee simulation model. A second goal was to determine the effect of lesion location within the knee and lesion size on accuracy and reliability. TYPE OF STUDY: Cross-sectional study of arthroscopic estimates of cartilage lesion size. METHODS: Three experienced arthroscopists performed 3 sets arthroscopic percent area estimates in 5 different plastic knees. Each knee had lesions drawn on 5 surfaces (patellar, medial and lateral femoral condyle, medial and lateral tibial plateaus). Accuracy and reliability were studied using Bland and Altman limits of agreement (LOA) and intraclass correlation coefficients. RESULTS: There was a strong tendency to overestimate lesion size by over 100% on the femoral and patellar surfaces. Intraobserver and interobserver reliabilities were generally poor. The range for the 95% LOA (+/- 1.96 standard deviation [SD] of the difference scores) between repeated measurements was almost 6 times the size of the lesion itself. Reliability of estimates was poorest for the largest lesions and worse at femoral, lateral tibial, and patellar sites. CONCLUSIONS: Assessments of arthroscopic measurements using LOA found that accuracy and reliability were generally poor, although results were better at the medial tibial plateau and for smaller lesions. In spite of these problems, arthroscopy remains a promising measurement tool because it permits physical assessment of cartilage integrity. This study sets the foundations for improvement in techniques of arthroscopic measurement of cartilage lesion size. PMID- 12627154 TI - Enveloping the tendon graft with periosteum to enhance tendon-bone healing in a bone tunnel: A biomechanical and histologic study in rabbits. AB - PURPOSE: Fixing and incorporating the tendon graft within the bone tunnel is a major concern when using grafts for ligament reconstruction. The periosteum contains multipotent stem cells and has the potential to form osteogenic and chondrogenic tissues. This study uses histologic and biomechanical analyses to examine the effect of periosteum on tendon-bone healing within a bone tunnel. TYPE OF STUDY: Experimental study in an animal model. METHODS: In this study, 36 adult New Zealand White rabbits were used. The long digitorum extensor tendon was transplanted into a bone tunnel of the proximal tibia. The periosteum from the proximal tibia was sutured on the surface of the tendon portion. The tendon was pulled through a drill-hole in the proximal tibia and attached to the medial aspect of the tibia. Histologic examination of the tendon-bone interface and biomechanical test for maximal pullout load were evaluated at 4, 8, and 12 weeks after operation. RESULTS: Histologic analysis of the tendon-bone interface showed a fibrous layer formed between the tendon and the bone by the periosteum. This layer became progressively integrated with the tendon and bone surface during the healing process. At 4 weeks, the cancellous bone lining in the bone tunnel was interdigitated with the fibrous interface tissue. At 8 weeks, progressive new bone grew into the interface fibrous layer. At 12 weeks, collagen fibers anchored to the bone and organization with fibrocartilage formation developed between the tendon and bone. Biomechanical testing revealed higher maximal pullout strength in the periosteum-enveloped group at all time points, with a statistically significant difference at 8 and 12 weeks. The periosteum-treated group had a higher interface strength-to-length ratio and significant increase at 8 weeks and 12 weeks. CONCLUSIONS: The histologic and biomechanical studies demonstrated that, if periosteum was sutured on the tendon that was transplanted within a bone tunnel, it resulted in a superior healing process and better healed strength. When doing ligament reconstruction with a tendon graft, the periosteum can be sutured to the graft to enhance tendon-bone healing. PMID- 12627155 TI - Knee stability and graft function following anterior cruciate ligament reconstruction: Comparison between 11 o'clock and 10 o'clock femoral tunnel placement. 2002 Richard O'Connor Award paper. AB - PURPOSE: To study how well an anterior cruciate ligament (ACL) graft fixed at the 10 and 11 o'clock positions can restore knee function in response to both externally applied anterior tibial and combined rotatory loads by comparing the biomechanical results with each other and with the intact knee. TYPE OF STUDY: Biomechanical experiment using human cadaveric specimens. METHODS: Ten human cadaveric knees (age, 41+/-13 years) were reconstructed by placing a bone patellar tendon-bone graft at the 10 and 11 o'clock positions, in a randomized order, and then tested using a robotic/universal force-moment sensor testing system. Two external loading conditions were applied: (1) 134 N anterior tibial load with the knee at full extension, 15 degrees, 30 degrees, 60 degrees, and 90 degrees of flexion, and (2) a combined rotatory load of 10 N-m valgus and 5 N-m internal tibial torque with the knee at 15 degrees and 30 degrees of flexion. The resulting kinematics of the reconstructed knee and in situ forces in the ACL graft were determined for each femoral tunnel position. RESULTS: In response to a 134-N anterior tibial load, anterior tibial translation (ATT) for both femoral tunnel positions was not significantly different from the intact knee except at 90 degrees of knee flexion as well as at 60 degrees of knee flexion for the 10 o'clock position. There was no significant difference in the ATT between the 10 and 11 o'clock positions, except at 90 degrees of knee flexion. Under a combined rotatory load, however, the coupled ATT for the 11 o'clock position was approximately 130% of that for the intact knee at 15 degrees and 30 degrees of flexion. For the 10 o'clock position, the coupled ATT was not significantly different from the intact knee at 15 degrees of flexion and approximately 120% of that for the intact knee at 30 degrees of flexion. Coupled ATT for the 10 o'clock position was significantly smaller than for the 11 o'clock position at 15 degrees and 30 degrees of flexion. The in situ force in the ACL graft was also significantly higher for the 10 o'clock position than the 11 o'clock position at 30 degrees of flexion in response to the same loading condition (70 +/- 18 N v 60 +/- 15 N, respectively). CONCLUSIONS: The 10 o'clock position more effectively resists rotatory loads when compared with the 11 o'clock position as evidenced by smaller ATT and higher in situ force in the graft. Despite the fact that ACL grafts placed at the 10 or 11 o'clock positions are equally effective under an anterior tibial load, neither femoral tunnel position was able to fully restore knee stability to the level of the intact knee. PMID- 12627156 TI - Should first time anterior shoulder dislocations be surgically stabilized? PMID- 12627157 TI - The analysis of articular cartilage after thermal exposure: "Is red really dead?". AB - Partial-thickness articular cartilage (PARC) lesions have no propensity to heal. Surgical techniques have focused on halting their progression. Thermal chondroplasty procedures have become controversial with regard to the viability of the remaining cartilage. This controversy has expanded to the way that articular cartilage is evaluated after treatment. In this article, evaluation of articular cartilage viability is reviewed. PMID- 12627158 TI - The use of intraoperative fluoroscopy to reduce femoral interference screw divergence during endoscopic anterior cruciate ligament reconstruction. AB - Femoral interference screw divergence can potentially have detrimental consequences in the rehabilitation of anterior cruciate ligament (ACL) reconstruction. Several biomechanical studies suggest that divergence angles greater than 15 degrees significantly decrease the pullout strength of the graft. Numerous techniques have been described in the literature that address this issue; however, the use of an image intensifier intraoperatively is not frequently discussed. We describe a technique in which fluoroscopy is used to confirm the proper position of the femoral interference screw at the time of the procedure, and therefore minimizes the incidence of significant screw divergence. Radiographic analysis of 62 patients who underwent endoscopic ACL reconstruction using bone-patellar tendon-bone autograft using this technique revealed significant divergence in only 3% of patients. PMID- 12627159 TI - Modified osteochondral autograft implantation for full- thickness articular cartilage lesions. AB - Full-thickness articular cartilage defects have been difficult to treat in patients with nonarthritic knees. A procedure is described to treat articular cartilage full-thickness lesions. Graft sites are chosen after appropriate treatment of the base of a grade IV lesion. Articular cartilage and bone are replaced into the graft site, promoting mesenchymal stem cell growth and cartilaginous coverage of the defect. Pathology, postoperative protocol, and some postoperative arthroscopic illustrations are included. This technique is simple and is associated with minimal donor-site morbidity. PMID- 12627160 TI - Posterior "back and forth" approach in arthroscopic surgery on the posterior knee compartments. AB - We report the feasibility, indications, and usefulness of an arthroscopic approach to the posterior knee compartments. We developed an arthroscopic approach to the posterior knee compartment involving use of a posteromedial portal and a posterolateral portal opposite each other. Each posterior portal is used in alternation for the arthroscope and instruments. The posterior portals are established using an original "back and forth" technique. The feasibility of the technique was evaluated on a cadaver. It allowed us to define safety rules to protect the vessels and nerves that course through the popliteal fossa. The approach provided a broader field of view compared with classical techniques. After removal of the septum dividing the posterior compartment, the synovial fold enclosing the posterior cruciate ligament and lining the upper and posterior parts of the posterior capsule was readily accessed, suggesting that this approach may be particularly valuable for total synovectomy. This was confirmed in 6 patients with villonodular synovitis, in whom the new approach was used in combination with arthroscopic anterior synovectomy. This technique allows removal of parts of the synovium that are difficult to access through conventional arthroscopic approaches. For total synovectomy, it can be used as an alternative to open posterior synovectomy, in combination with arthroscopic anterior synovectomy. PMID- 12627161 TI - Iliotibial band Z-lengthening. AB - Iliotibial band friction syndrome (ITBFS) is a common overuse injury reported to afflict 1.6% to 12% of runners. It results from an inflammatory response secondary to excessive friction that occurs between the lateral femoral epicondyle and the iliotibial band. Initial treatments include rest, anti inflammatory medication, modalities (ice or heat), stretching, physical therapy, and possibly a cortisone injection. In recalcitrant cases of ITBFS, surgery has been advocated. This report describes a surgical technique of Z-lengthening of the iliotibial band in patients presenting with lateral knee pain localized to the iliotibial band at the lateral femoral epicondyle and Gerdy's tubercle who failed all nonoperative efforts. PMID- 12627162 TI - A modified Mason-Allen technique for rotator cuff repair using suture anchors. AB - An adequate restoration of the muscle-tendon-bone unit is essential for a successful outcome after rotator cuff reconstruction. We describe a suture grasping technique for rotator cuff repair using suture anchors that can be performed either arthroscopically or during open rotator cuff refixation. The technique we use is a combination of a mattress and a single suture, thus representing the principles of a Mason-Allen suture technique. The modified Mason Allen technique for suture anchor repair is easy to perform and provides excellent initial fixation strength allowing durable osteofibroblastic integration of the reinserted cuff. PMID- 12627163 TI - The comma sign: An arthroscopic guide to the torn subscapularis tendon. AB - Subscapularis tears are becoming increasingly recognized as a cause of shoulder pain and disability. However, identifying the subscapularis tendon stump is often difficult during repair of chronic, retracted subscapularis tears that are scarred to the deltoid fascia. The authors have found the "comma sign," an arc formed by a portion of the superior glenohumeral ligament/coracohumeral ligament complex, to be a useful marker of the superolateral corner of the torn subscapularis tendon. PMID- 12627165 TI - A motor learning strategy reflects neural circuitry for limb control. AB - During motor skill acquisition, the brain learns a mapping between intended limb motion and requisite muscular forces. We propose that regions where sensory and motor representations overlap are crucial for motor learning. In primary motor cortex, for example, cells that modulate their activity for motor actions at a joint tend to receive input from that same portion of the periphery. We predict that this correspondence reflects a default strategy--a Bayesian prior--in which subjects tend to associate loads at a joint with motion at that joint (local sensorimotor association) when there is ambiguity regarding the nature of the load. As predicted, we found that in the presence of uncertainty, humans inappropriately generalized elbow loads as though they were based on elbow velocity. Generalization improved when we reduced uncertainty by decreasing coupling between elbow velocity and load during training. These results illustrate a key link between motor learning and the underlying neural circuitry. PMID- 12627164 TI - Neuronal correlates of perception in early visual cortex. AB - We used functional magnetic resonance imaging (fMRI) to measure activity in human early visual cortex (areas V1, V2 and V3) during a challenging contrast-detection task. Subjects attempted to detect the presence of slight contrast increments added to two kinds of background patterns. Behavioral responses were recorded so that the corresponding cortical activity could be grouped into the usual signal detection categories: hits, false alarms, misses and correct rejects. For both kinds of background patterns, the measured cortical activity was retinotopically specific. Hits and false alarms were associated with significantly more cortical activity than were correct rejects and misses. That false alarms evoked more activity than misses indicates that activity in early visual cortex corresponded to the subjects' percepts, rather than to the physically presented stimulus. PMID- 12627166 TI - Superior colliculus encodes distance to target, not saccade amplitude, in multi step gaze shifts. AB - The superior colliculus (SC) is important for generating coordinated eye-head gaze saccades. Its deeper layers contain a retinotopically organized motor map in which each site is thought to encode a specific gaze saccade vector. Here we show that this fundamental assumption in current models of collicular function does not hold true during horizontal multi-step gaze shifts in darkness that are directed to a goal and composed of a sequence of gaze saccades separated by periods of steady fixation. At the start of a multi-step gaze shift in cats, neural activity on the SC's map was located caudally to encode the overall amplitude of the gaze displacement, not the first saccade in the sequence. As the gaze shift progressed, the locus of activity moved to encode the error between the goal and the current gaze position. Contrary to common belief, the locus of activity never encoded gaze saccade amplitude, except for the last saccade in the sequence. PMID- 12627167 TI - Perceptual interference supports a non-modular account of face processing. AB - The perception of faces and of nonface objects share common early visual processing stages. Some argue, however, that the brain eventually processes faces separately from other objects, within a domain-specific module dedicated to face perception. This apparent specialization for faces could, alternatively, result from people's expertise with this category of stimuli. Here we used behavioral and electrophysiological measures of interference to address the functional independence of face and object processing. If the expert processing of faces and cars depend on common mechanisms related to holistic perception (obligatory processing of all parts), then for human subjects who are presumed to be face experts, car perception should interfere with concurrent face perception. Furthermore, such interference should increase with greater expertise in car identification, and indeed this is what we found. Event-related potentials (ERPs) suggest that this interference arose from perceptual processes contributing to the holistic processing of both objects of expertise and faces. PMID- 12627168 TI - Genetic engineering of an immunotoxin to eliminate pulmonary vascular leak in mice. AB - Vascular leak syndrome is a major and often dose-limiting side effect of immunotoxins and cytokines. We postulated that this syndrome is initiated by damage to vascular endothelial cells. Our earlier studies identified a three amino acid motif that is shared by toxins, ribosome-inactivating proteins, and interleukin-2, all of which cause this problem. We have now generated a panel of recombinant ricin A chains with mutations in this sequence or in amino acids flanking it in the three-dimensional structure. These have been evaluated alone and as immunotoxins for activity, ability to induce pulmonary vascular leak in mice, pharmacokinetics, and activity in tumor-xenografted mice. One mutant was comparable to the ricin A chain used before in all respects except that it did not cause vascular leak at the same dose and, when used as an immunotoxin, was more effective in xenografted SCID mice. PMID- 12627169 TI - A transformation method for obtaining marker-free plants of a cross-pollinating and vegetatively propagated crop. AB - It is generally thought that transformation of plant cells using Agrobacterium tumefaciens occurs at a very low frequency. Therefore, selection marker genes are used to identify the rare plants that have taken up foreign DNA. Genes encoding antibiotic and herbicide resistance are widely used for this purpose in plant transformation. Over the past several years, consumer and environmental groups have expressed concern about the use of antibiotic- and herbicide-resistance genes from an ecological and food safety perspective. Although no scientific basis has been determined for these concerns, generating marker-free plants would certainly contribute to the public acceptance of transgenic crops. Several methods have been reported to create marker gene-free transformed plants, for example co-transformation, transposable elements, site-specific recombination, or intrachromosomal recombination. Not only are most of these systems time-consuming and inefficient, but they are also employed on the assumption that isolation of transformants without a selective marker gene is not feasible. Here we present a method that permits the identification of transgenic plants without the use of selectable markers. This strategy relies on the transformation of tissue explants or cells with a virulent A. tumefaciens strain and selection of transformed cells or shoots after PCR analysis. Incubation of potato explants with A. tumefaciens strain AGL0 resulted in transformed shoots at an efficiency of 1-5% of the harvested shoots, depending on the potato genotype used. Because this system does not require genetic segregation or site-specific DNA-deletion systems to remove marker genes, it may provide a reliable and efficient tool for generating transgenic plants for commercial use, especially in vegetatively propagated species like potato and cassava. PMID- 12627170 TI - Identification of co-regulated genes through Bayesian clustering of predicted regulatory binding sites. AB - The identification of co-regulated genes and their transcription-factor binding sites (TFBS) are key steps toward understanding transcription regulation. In addition to effective laboratory assays, various computational approaches for the detection of TFBS in promoter regions of coexpressed genes have been developed. The availability of complete genome sequences combined with the likelihood that transcription factors and their cognate sites are often conserved during evolution has led to the development of phylogenetic footprinting. The modus operandi of this technique is to search for conserved motifs upstream of orthologous genes from closely related species. The method can identify hundreds of TFBS without prior knowledge of co-regulation or coexpression. Because many of these predicted sites are likely to be bound by the same transcription factor, motifs with similar patterns can be put into clusters so as to infer the sets of co-regulated genes, that is, the regulons. This strategy utilizes only genome sequence information and is complementary to and confirmative of gene expression data generated by microarray experiments. However, the limited data available to characterize individual binding patterns, the variation in motif alignment, motif width, and base conservation, and the lack of knowledge of the number and sizes of regulons make this inference problem difficult. We have developed a Gibbs sampling-based Bayesian motif clustering (BMC) algorithm to address these challenges. Tests on simulated data sets show that BMC produces many fewer errors than hierarchical and K-means clustering methods. The application of BMC to hundreds of predicted gamma-proteobacterial motifs correctly identified many experimentally reported regulons, inferred the existence of previously unreported members of these regulons, and suggested novel regulons. PMID- 12627171 TI - Site-specific gene targeting in mouse embryonic stem cells with intact bacterial artificial chromosomes. AB - Homologous recombination in Escherichia coli simplifies the generation of gene targeting constructs for transduction into mouse embryonic stem (ES) cells. Taking advantage of the extensive homology provided by intact bacterial artificial chromosomes (BACs), we have developed an efficient method for preparing targeted gene disruptions in ES cells. Correctly integrated clones were identified by a simple screening procedure based on chromosomal fluorescence in situ hybridization (FISH). To date, five mutant lines have been generated and bred to homozygosity by this approach. PMID- 12627172 TI - Engineering the mouse genome with bacterial artificial chromosomes to create multipurpose alleles. AB - The mouse is the leading vertebrate model because its genome can be altered by both random transgenesis and homologous recombination with targeting constructs. Both approaches have been hindered by the size and site limitations implicit in conventional Escherichia coli DNA-engineering methods. Homologous recombination in E. coli, or 'recombineering', has overcome these limitations for bacterial artificial chromosome (BAC) transgenesis. Here we applied Red/ET recombineering (using the lambda Redalpha/Redbeta recombinase pair) to generate a 64 kilobase targeting construct that carried two selectable cassettes permitting the simultaneous mutation of the target gene, Mll, at sites 43 kb apart in one round of mouse embryonic stem (ES) cell targeting. The targeting frequency after dual selection was 6%. Because the two selectable cassettes were flanked by FRT or loxP sites, three more alleles can be generated by site-specific recombination. Our approach represents a simple way to introduce changes at two or more sites in a genetic locus, and thereafter generate allele combinations. The size of BAC templates offers new freedom for the design of targeting constructs. Combined with the use of two selectable cassettes placed far apart, BAC-based targeting constructs may be applicable to tasks such as regional exchanges, deletions, and insertions. PMID- 12627173 TI - Lean body mass in preschool aged urban children in India: gender difference. AB - OBJECTIVE: To estimate lean body mass (LBM) in preschool aged boys and girls in India and explore gender difference. DESIGN: Crossectional. SETTING: Immunization clinic of a charitable Government General Hospital in Kolkata, India. SUBJECTS: Two-hundred and forty-five children (147 boys and 98 girls) aged 1-5 y from among the urban poor were admitted in the study between July 1999 and December 2000. Children with acute or chronic illness or congenital malformation were excluded. METHOD: Length/height to the nearest 0.1 cm, weight to the nearest 10 g and total body resistance using multifrequency bioelectrical (Xitron 4000B) impedance analyzer (BIA) at 50 kHz were measured. Their nutritional status was compared with National Center for Health Statistics (NCHS) median data and lean body mass (LBM) was calculated using anthropometry and BIA equations. The groups were compared using analysis of variance and multiple linear regression. RESULTS: Girls were more stunted (P<0.001) and underweight (P<0.047), while the degree of wasting was similar. Mean LBM percentage was higher in boys compared with girls by anthropometry (P<0.001) and BIA (P<0.005), which persisted after adjusting for age. With increasing age, LBM percentage declined in girls (P<0.02) in contrast to reference girls, in whom it increased. In boys LBM percentage increased with age as is in reference boys. CONCLUSIONS: In addition to the girls being more stunted and underweight, LBM% decreased in girls with increasing age but steadily increased in boys, suggesting hidden deprivation of female children. PMID- 12627174 TI - Computerization of a dietary history interview in a running cohort; evaluation within the Amsterdam Growth and Health Longitudinal Study. AB - OBJECTIVE: In nutritional research, a growing interest in the use of computer assisted cross-check dietary history interview methods exists in order to improve cost-effectiveness. The introduction of such a method in an ongoing longitudinal study was evaluated with special emphasis on the effect on interviewer bias. DESIGN: A study for the interviewer bias within and the agreement between a previously used paper-based face-to-face cross-check dietary history interview method and a newly developed interviewer-administered computer-assisted version of this interview method. SUBJECTS: The interviewer bias of 436 face-to-face interviews is compared with that of 352 computer-assisted interviews. A subset of 82 subjects underwent a face-to-face interview at the mean age of 27 and 32 y and a computer-assisted interview at their mean age of 36 y. Energy, three macronutrients (protein, fat and carbohydrate), two micronutrients (calcium and iron) and alcohol intakes obtained by these three measurements are compared to analyse the agreement between the two interview methods. RESULTS: ANOVA showed no interviewer bias for all seven analysed nutrients within the data from the computer-assisted interview, while for the face-to-face interview method, several nutrients varied significantly among the interviewers. Five different measures, used to analyse the agreement (differences, Pearson's correlation, ICC, square weighted kappa and Bland-Altman plots), showed no relevant differences between the two cross-check dietary history interview methods. CONCLUSIONS: It is concluded that the computer-assisted interview caused a reduction of interviewer bias and is of similar quality to the face-to-face interview method. Computerization of a paper-based interview can be implemented in a running cohort if a change in method is unavoidable. PMID- 12627175 TI - Differences in body composition between Singapore Chinese, Beijing Chinese and Dutch children. AB - OBJECTIVES: To compare the relationship between body mass index (BMI) and body fat percentage (BF%) in children of different ethnic background. DESIGN: Cross sectional observational study. SETTINGS: The study was performed in three different locations, Singapore, Beijing and Wageningen (The Netherlands). SUBJECTS: In each centre 25 boys and 25 girls, aged 7-12 y, were selected. They were matched on age, sex and body height. METHODS: Body weight and body height was measured following standardized procedures. The body mass index (BMI) was calculated as weight/height squared (kg/m(2)). Body fat was measured by densitometry in Beijing and Wageningen and by dual energy X-ray absorptiometry (DXA) in Singapore. The DXA measurements in Singapore were validated against densitometry. RESULTS: There were no significant differences in BF% or BMI within each gender group across the three study sites. However, after controlling for (non-significant) differences in age and BF%, the Singapore children had a lower (mean+/-s.e.) BMI (15.6+/-0.3) than the Beijing 17.6+/-0.3) and Wageningen (16.9+/-0.3) children. For the same BMI, age and sex the Singapore children had a significant higher BF% (24.6+/-0.7) than the Beijing (19.2+/-0.8) and Wageningen (20.3+/-0.7) children. CONCLUSIONS: The study strongly suggests that the relationship between BF% and BMI (or weight and height) is different among children of different ethnic background. Consequently growth charts and BMI cut off points for underweight, overweight and obesity in children may have to be ethnic-specific. PMID- 12627176 TI - Measurement of vitamin E metabolites by high-performance liquid chromatography during high-dose administration of alpha-tocopherol. AB - METHOD: alpha-Tocopherol and gamma-tocopherol are metabolized into 2,5,7,8 tetramethyl-2-(2'-carboxyethyl)-6-hydroxychroman (alpha-CEHC) and 2,7,8-trimethyl 2-(2'-carboxyethyl)-6-hydroxychroman (gamma-CEHC), respectively. We analyzed alpha- and gamma-CEHC concentrations in human serum and urine by high-performance liquid chromatography during administration of alpha-tocopherol. Fourteen healthy adult male volunteers received 1,200 IU per day of RRR-alpha-tocopherol for 28 days. Blood and urine samples were obtained on days 0, 14, 28, and 56. RESULTS: During alpha-tocopherol administration, the plasma gamma-tocopherol concentration decreased significantly, but there was marked elevation of the alpha-tocopherol concentration. Increased concentration of alpha-CEHC and gamma-CEHC in both serum and urine indicated the acceleration of vitamin E metabolism. CONCLUSION: High dose administration of alpha-tocopherol caused an increase of gamma-tocopherol metabolism, which might have caused a decrease of the plasma gamma-tocopherol concentration. PMID- 12627177 TI - Bioavailability of soluble oxalate from tea and the effect of consuming milk with the tea. AB - OBJECTIVES: To measure the availability of oxalate normally extracted when making tea from two commercially available black teas bought from a supermarket in Christchurch, New Zealand in July 2001. DESIGN, SUBJECTS AND INTERVENTION: A randomized double crossover study. Six students and four staff consumed six cups of each brand of tea both with and without added milk over a 24 h period. A total urine collection was taken for the initial 6 h followed by a further 18 h. The oxalate content of the urine voided was measured using an enzyme kit method and the availability of the soluble oxalate consumed was measured for the 6 h and the total 24 h sample. SETTING: University campus. RESULTS: The mean soluble oxalate content of black tea in the two different commercial tea bags was respectively 6.1 and 6.3 mg soluble oxalate/g tea. The mean availability of the oxalate extracted from tea measured over a 6 h period ranged from 1.9 to 4.7% when tea was consumed without milk. The availability of the soluble oxalate from tea ranged from -3.0 to 2.3% for each of the two brands of tea investigated over a 24 h period. CONCLUSION: These studies show that consuming black tea on a daily basis will lead to a moderate intake of soluble oxalate each day, however the consumption of tea with milk on a regular basis will result in the absorption of very little oxalate from tea. PMID- 12627178 TI - Obesity in a cohort of black Jamaican children as estimated by BMI and other indices of adiposity. AB - OBJECTIVE: To examine the relationships of body mass index (BMI) to obesity indices derived from anthropometry and to determine tracking of overweight between late childhood and early adolescence, in a cohort of children with mixed nutritional history. We also compared identification of overweight children using The International Obesity Task Force (IOTF) BMI cut-off points with skinfolds. DESIGN: Prospective study. SETTING: Kingston, Jamaica. SUBJECTS: A total of 306 children examined at 7-8 y and at 11-12 y. MEASUREMENTS: Triceps (TSF) and subscapular skinfolds (SSF), height and weight were measured. The sum of the skinfolds (sum SF), BMI, percentage body fat (%fat) and fat mass (FM) were calculated. Pubertal stage was assessed at 11-12 y. RESULTS: Overweight increased from 3.5 to 9.5% over the follow-up period. BMI was better correlated with the other indices of adiposity in girls and in the older age group. BMI tracking over follow up was high. In regression analysis BMI explained 52 and 61% of the variance in FM in boys and girls at 7-8 y. This increased to 69% in both sexes at 11-12 y. Using the IOTF cut-off points BMI had low sensitivity to identify children >85th percentile of the NHANES references for SSF. The sensitivity for those assessed by TSF and sum SF was higher, but between 14 and 30% of the children were misclassified. The specificity of BMI was high. CONCLUSIONS: Adiposity increased over follow-up. Although the cohort remained relatively lean BMI rank among the fattest children was maintained. Girls were fatter than boys, reflecting adult obesity patterns. Children identified as overweight by the IOTF BMI cut-off points are likely to have high body fatness. However the BMI cut-off points may not identify many children with high body fatness. PMID- 12627179 TI - The glycaemic index of dates and date/yoghurt mixed meals. Are dates 'the candy that grows on trees'? AB - OBJECTIVE: To determine the glycaemic index of one variety of date, alone and in mixed meals with plain full-milk yoghurt. DESIGN: An experimental study involving the measurement of the glycaemic responses of participants to the ingestion of dates, alone and with yoghurt. SETTING: Within the Department of Family Medicine, Faculty of Medicine and Health Sciences, UAE University. SUBJECTS: Healthy male and female volunteers aged 32-67 y. INTERVENTION: Glycaemic responses to date and date/yoghurt meal ingestion were plotted for each subject. Glycaemic indexes were calculated as the ratios of the incremental areas under these response curves to those for glucose ingestion. Results are presented using means and standard deviations. Glycaemic indexes were compared using paired t-tests. RESULTS: Mean glycaemic indexes of the dates were 47.2, 45.3, 35.5, 37.3, 28.9 for rutab, traditionally stored, commercial, rutab/yoghurt and commercial tamer/yoghurt preparations, respectively. There was a significant difference between the results for rutab vs commercial tamer dates (P<0.05), but other comparisons failed to reach statistical significance. CONCLUSIONS: Khalas dates, when eaten alone or in mixed meals with plain yoghurt have low glycaemic indexes. The consumption of dates may be of benefit in glycaemic and lipid control of diabetic patients. The consumption of dates in mixed meals with yoghurt appears to have, at most, a minimal effect on the glycaemic index. PMID- 12627180 TI - Nutritional deficiencies in German middle-class male alcohol consumers: relation to dietary intake and severity of liver disease. AB - OBJECTIVE: The purpose of the present study was to compare the nutrient intake and the nutritional status between German middle-class alcohol consumers and non drinkers. DESIGN: Cross-sectional study using patients with different stages of alcoholic liver disease (ALD) and healthy volunteers. SETTING: Southern Germany. SUBJECTS: Seventy-six hospitalized German middle-class alcohol consumers with different stages of alcoholic liver disease (ALD) and 22 healthy control subjects. METHODS: Subjects and controls were nutritionally assessed and mineral and vitamin content was measured in blood and urine. RESULTS: When compared with controls, alcohol consumers had significantly higher intakes of total calories, but intake of non-alcoholic calories did not differ between groups (P<0.05). Among drinkers, there was a decrease in percentage of energy derived from protein and fat and a significant increase in carbohydrates (P<0.05). With the exception of vitamin E, micronutrient intake of alcoholics was equal to that of controls; however, blood vitamin (vitamin C, retinol, lycopene, alpha- and gamma-carotene) and trace element (selenium, zinc) concentrations of alcohol-drinking patients were lower than those of non-drinkers. CONCLUSION: From the results of this study it is concluded that in German middle-class male alcohol consumers the status of several micronutrients is disturbed, although dietary intake hardly differs from that in non-alcoholic controls. PMID- 12627181 TI - Mediterranean diet, but not red wine, is associated with beneficial changes in primary haemostasis. AB - OBJECTIVE: (1) To compare the effect of an alcohol-free Mediterranean-type diet (MD) and a high-fat diet (HFD) on variables of primary haemostasis (bleeding time, plasma von Willebrand factor and platelet aggregation/secretion). (2) To test whether red wine supplementation modified these variables, independently of the diet. DESIGN, SUBJECTS AND INTERVENTION: Controlled prospective intervention study. Two groups, each consisting of 21 healthy male university students (22+/ 3.4 y), received either MD or HFD during 90 days. Between days 30 and 60, both diets were supplemented with 240 ml/day of red wine. Baseline (T0) and T30, T60 and T90-day samples were drawn. Bleeding time was measured before (day 30) and after (day 60) wine supplementation. No drop out from the study was experienced. SETTING: University campus and outpatient nutrition clinic. RESULTS: All baseline (day 0) variables did not differ significantly between study groups. On day 30, individuals on MD had significantly higher levels of plasma beta-carotene, folate, ascorbate, and eicosapentaenoic acid in plasma lipid fractions, than those on HFD. Total plasma cholesterol, HDL and LDL did not change significantly in either study group at any time point. After 30 days on each diet, individuals on MD had longer bleeding time (BT) than those on HFD (7.6+/-2.8 vs 5.8+/-1.7 min; P=0.017). BT did not change significantly after I month of wine supplementation (7.1+/-2.0 vs 5.5+/-2.0 min, respectively). Plasma von Willebrand factor (vWF : Ag) on day 0 was 89+/-40 and 111+/-70% in MD and HFD groups, respectively (P=0.21). These values did not change significantly at 30, 60 or 90 days. MD intake was associated with an increase in platelet serotonin secretion (P=0.02) and a marginal increase in platelet aggregation after stimulation with epinephrine (P=0.07). Wine intake resulted in a marginal decrease in platelet (14)C-5-HT secretion with 4 micro M ADP (P=0.07). However, both platelet aggregation and secretion were consistently increased when using collagen as agonist (1 and 2 micro g/ml, P=0.01). CONCLUSION: The longer BT in individuals on MD, obtained independently of red wine, denotes less interaction of platelets with the vascular wall, which could be beneficial from the point of view of cardiovascular (CV) risk. This effect is not explained by changes in the measured haemostatic determinants of BT (plasma vWF, ex vivo platelet function), and might be attributed to other as yet unknown vascular factors. Moderate consumption of red wine results in a significant increase in ex vivo platelet aggregation and secretion after stimulation with collagen. This observation contradicts previous reports, although further studies are required to elucidate the influence of this finding on CV risk. PMID- 12627182 TI - Factors contributing to variation in lipoprotein (a) in Melbourne Anglo-Celtic population. AB - AIM: The purpose of this report is to survey the factors contributing to variation in lipoprotein(a) (Lp(a)) in a population-based sample of Anglo-Celtic Melburnians. RESULTS: The plasma Lp(a) levels were highly skewed towards low levels in this population, with a median of 156 mg/l and a mean of 262 mg/l. Approximately 33% had plasma Lp(a) above the threshold value of 300 mg/l, while 35% had Lp(a) levels below 100 mg/l. The most commonly occurring phenotype was apo(a) S3. In this phenotype, Lp(a) concentrations ranged from 10 to 596 mg/l. Lp(a) was consistently associated with diastolic blood pressure, systolic blood pressure, total protein, albumin and nitrogen excretion in the 40-60 y age group. Multiple stepwise regression analyses, in non-dietary factors, were used to explain about 13% of the variance in Lp(a) (19% in men and 23% in women). Remarkably, in the <40 y age group, non-dietary factors may account for 86% of the variance in Lp(a) and dietary factors, analysed separately, 46%. Thus, although Lp(a) is mainly genetically determined, there are clearly other factors which contribute to variations in Lp(a) concentrations. PMID- 12627183 TI - An evaluation of the sensitivity and specificity of energy expenditure measured by heart rate and the Goldberg cut-off for energy intake: basal metabolic rate for identifying mis-reporting of energy intake by adults and children: a retrospective analysis. AB - OBJECTIVE: To identify adults and children as under- (UR), acceptable (AR), or over-reporters (OR) of energy intake (EI) using energy expenditure measured by doubly labelled water (DLW) (EE(DLW)), and to use this as a reference to determine the sensitivity and specificity of (i) EE measured by heart rate (EE(HR)), and (ii) the Goldberg cut-off technique for classifying subjects into the same categories. DESIGN: Retrospective analysis of a dataset comprising concurrent measurements of EE(DLW), EE(HR), basal metabolic rate (BMR), and EI by weighed record (EI(WR)) on 14 adults and 36 children. EI by diet history (EI(DH)) was also measured in the children only. EI(WR):EE(DLW) provided the reference definition of subjects as UR, AR or OR. Three strategies for classifying mis reporters based on EE(HR) and Goldberg cut-offs were then explored. Sensitivity and specificity were calculated respectively as the proportion of UR and non-UR correctly identified. RESULTS: Approximately 80% of all subjects were AR. For EI(WR) and EI(DH) respectively, the sensitivity of EE(HR) was 0.50 and 1.00, and specificity was 0.98 and 1.00. Although designating subjects as having low, medium or high activity levels (EE(HR):BMR(meas)) and calculating cut-offs based on appropriate WHO physical activity level PALs did not change sensitivity, specificity dropped to 0.98 (EI(WR)) and 0.97 (EI(DH)). Cut-offs based on a PAL of 1.55 reduced sensitivity to 0.33 (EI(WR)) and 0.00 (EI(DH)), but specificity remained unchanged. The sensitivity of all cut-offs based on physical activity level (PALs) for EI(WR) was 0.50 (adults) and 0.25 (children). CONCLUSIONS: If the precision of EE(HR) was improved, it may be useful for identifying mis reporters of EI. PMID- 12627184 TI - Fiber intake and risk of nonfatal acute myocardial infarction. AB - OBJECTIVE: To investigate the association between fiber intake and risk of acute myocardial infarction (AMI), also according to type and source of fiber, in a Mediterranean country. DESIGN: Hospital-based case-control study. SETTING: Northern Italy. SUBJECTS: A total of 507 cases of first nonfatal AMI and 478 controls in hospital for acute conditions. INTERVENTIONS: Subjects were interviewed with a questionnaire that included a validated food frequency section. Odds ratios (OR) were obtained using multiple logistic regression, and adjusted for several recognized risk factors for AMI. Fiber was measured as non starch polysaccharides. RESULTS: Compared with the lowest one, the OR in the highest tertile was 0.72 for total fiber, 0.64 for soluble fiber, 0.77 for total insoluble fiber, 0.71 for cellulose, 0.81 for insoluble non-cellulosic polysaccharides, 0.82 for vegetable fiber, 0.64 for fruit fiber and 1.11 for cereal fiber, and the estimates were statistically significant for soluble and fruit fiber. When further adjusted for beta-carotene, vitamin C and vitamin E intake, the fruit fiber still showed the strongest inverse relation, although the association was no longer significant. The protective effect of fiber was more marked in, or restricted to, subjects with other AMI risk factors, such as smokers, diabetics and hypertensives. CONCLUSIONS: Though an inverse association between fiber intake and AMI risk appears established, the causality of this association is still open to debate. In this population, cereal fiber derives chiefly from refined grains, and this may explain the lack of protection by this type of fiber. PMID- 12627185 TI - Prevalence of overweight and obesity among Bahraini adolescents: a comparison between three different sets of criteria. AB - OBJECTIVE: The aim of this study is to determine the prevalence of overweight and obesity among Bahraini adolescents using three different sets of criteria/standards. DESIGN: Cross-sectional prevalence study. SETTING: Intermediate and secondary schools in Bahrain. SUBJECTS: The study included a population-representative sample of 506 Bahraini students (249 males and 257 females) between 12 and 17 y of age. The sample was selected using multistage stratified random sampling technique. MEASUREMENT: Anthropometric measurements including weight, height and triceps and subscapular skinfolds were taken on the adolescents. Age was verified against school records. To minimize inter-observer error, weight and height were taken by one person while skinfold was taken by two trained persons (one for each sex). RESULTS: The overall prevalence of obesity among Bahraini boys and girls was high, especially in girls. Obesity was highest (21% in males and 35% in females) when the WHO recommended criteria of BMI for age and skinfolds for age percentiles were applied and lowest (15% in boys and 18% in girls) when the age and sex specific BMI cut-off values of Cole et al were used. Compared with those of WHO criteria, estimates of overweight and obesity prevalence obtained with Must et al and Cole et al were generally close. CONCLUSIONS: Our data revealed a much higher prevalence rate of obesity in the Bahraini adolescent population than was previously reported, especially among girls. The BMI reference values of Must et al and that of Cole et al gave relatively similar estimates and appear to be more practical for use in surveys aimed at estimating the prevalence of overweight and obesity among adolescents than the WHO recommended composite criteria. PMID- 12627186 TI - Determination of the glycaemic index of foods: interlaboratory study. AB - OBJECTIVE: Practical use of the glycaemic index (GI), as recommended by the FAO/WHO, requires an evaluation of the recommended method. Our purpose was to determine the magnitude and sources of variation of the GI values obtained by experienced investigators in different international centres. DESIGN: GI values of four centrally provided foods (instant potato, rice, spaghetti and barley) and locally obtained white bread were determined in 8-12 subjects in each of seven centres using the method recommended by FAO/WHO. Data analysis was performed centrally. SETTING: University departments of nutrition. SUBJECTS: Healthy subjects (28 male, 40 female) were studied. RESULTS: The GI values of the five foods did not vary significantly in different centres nor was there a significant centrexfood interaction. Within-subject variation from two centres using venous blood was twice that from five centres using capillary blood. The s.d. of centre mean GI values was reduced from 10.6 (range 6.8-12.8) to 9.0 (range 4.8-12.6) by excluding venous blood data. GI values were not significantly related to differences in method of glucose measurement or subject characteristics (age, sex, BMI, ethnicity or absolute glycaemic response). GI values for locally obtained bread were no more variable than those for centrally provided foods. CONCLUSIONS: The GI values of foods are more precisely determined using capillary than venous blood sampling, with mean between-laboratory s.d. of approximately 9.0. Finding ways to reduce within-subject variation of glycaemic responses may be the most effective strategy to improve the precision of measurement of GI values. PMID- 12627187 TI - Reduction in plasma total homocysteine through increasing folate intake in healthy individuals is not associated with changes in measures of antioxidant activity or oxidant damage. AB - BACKGROUND: Various mechanisms have been proposed to explain the association between plasma total homocysteine (tHcy) and risk of cardiovascular disease, including oxidative activity of homocysteine. OBJECTIVE: To explore the putative role of reactive oxygen species in the association between plasma tHcy and risk of cardiovascular disease in healthy individuals. DESIGN: A double-blind, placebo controlled crossover intervention to increase folate intake through diet (increased consumption of folate-rich foods) and supplement (400 micro g folic acid) was carried out in 126 healthy men and women. Measurements were made of antioxidant activity in red blood cells and plasma, and products of oxidant damage in plasma. RESULTS: Diet and supplement-based interventions led to an increase in measures of folate status and a reduction in plasma tHcy. This was not associated with any significant change in measures of antioxidant activity (plasma and red blood cell glutathione peroxidase activity and red blood cell superoxide dismutase activity) or oxidant damage (plasma malondialdehyde), although an improvement in plasma total antioxidant capacity just failed to reach significance. CONCLUSIONS: In healthy individuals lowering plasma tHcy does not have any functional implications regarding oxidative damage. PMID- 12627188 TI - Blood-pressure-lowering effect of a novel fermented milk containing gamma aminobutyric acid (GABA) in mild hypertensives. AB - OBJECTIVE: To study the effect of a new fermented milk product containing GABA (FMG) on the blood pressure (BP) of patients with mild hypertension. DESIGN: A randomized, placebo-controlled, single-blind trial. SETTING: The study was carried out at the outpatient clinic of the Cardiovascular Disease Center, Tokyo Metropolitan Police Hospital, Japan. SUBJECTS: The study population comprised 39 mildly hypertensive patients (16 women and 23 men) aged 28-81 y (mean, 54.2 y). INTERVENTIONS: The study consisted of a 12-week period of daily intake of FMG or placebo (weeks 1-12) followed by 2 weeks of no intake (weeks 13 and 14). We measured the peripheral BP and heart rate of seated patients at weeks 0, 2, 4, 8, 12 and 14. Routine blood study and urinalysis were performed before and after the intake. RESULTS: There was a significant decrease of BP within 2 or 4 weeks, and it remained decreased throughout the 12-week intake period. For the FMG recipients, the mean decrease after 12 weeks was 17.4+/-4.3 mmHg in the systolic BP (SBP) and 7.2+/-5.7 mmHg in the diastolic BP (DBP). Both of these values differed statistically from baseline levels (P<0.01), and the SBP of the FMG group differed from the placebo group (P<0.05). Heart rate, body weight, hematological and blood chemistry variables, and urinalysis results (glucosuria and proteinuria) did not vary both groups throughout the study. CONCLUSION: FMG may contribute to lowering BP in mildly hypertensive people. PMID- 12627189 TI - Fatty acid abnormalities in chronic pancreatitis: effect of concomitant diabetes mellitus. AB - OBJECTIVE: Patients with chronic pancreatitis suffer from malabsorption and nutritional deficiencies. However there is little data available concerning the fatty acid profile in chronic pancreatitis. Diabetes mellitus, a common complication of this disease, could interfere with the metabolism of fatty acids. SUBJECTS: We therefore compared the fatty acid composition of LDL from four groups of male patients with (a) chronic pancreatitis without diabetes (ND-CP; n=12), (b) diabetes secondary to chronic pancreatitis and insulin-treated (CP-D; n=35); (c) type 1 diabetes (n=25); and (d) controls (n=20). RESULTS: The patients in both groups of chronic pancreatitis (ND-CP and CP-D) had lower mean values for linoleic acid than that seen in the type 1 DM and control groups, whereas monounsaturated fatty acids (MUFA; 18 : 1(n-9) and (16 : 1(n-7)) were significantly increased in these two groups (ND-CP and CP-D). Docosa-hexaenoic acid (22 : 6(n-3)) was significantly decreased in the CP-D group (P>0.05), a response that could be explained by the effects of diabetes mellitus and by selenium deficiency. In this way, diabetes was associated with a decrease in the docosa-hexaenoic-acid (22 : 6(n-3); r=0.30, P=0.005), and selenium was correlated with DHA (r=0.28, P=0.029) and with the 22 : 6(n-3)/20 : 5(n-3) ratio (evaluating the delta 4 desaturation); r=0.31, P=0.022), independently of the diabetes effect. Selenium was negatively correlated with 20 : 4(n-6)/20 : 3(n-6) ratio (evaluating the delta 5 desaturase; r=-0.30; P=0.025). These results suggest that these two factors may have a role in the regulation of the desaturation process. If we consider that a ratio of 16 : 1(n-7)/18 : 2(n-6) greater than 0.086 in plasma indicates an EFAn-6 deficiency, 40% of our CP patients, 57.6% of CP-D patients and 13.6% of type 1 DM patients were involved. CONCLUSIONS: The consequences of these deficiencies are not evaluated in this disease. However, correction of the fundamental deficiencies in essential fatty acids and in selenium seems desirable in chronic pancreatitis. PMID- 12627190 TI - Challenges for the 'Rep' body. PMID- 12627191 TI - Prolonged paraesthesia. PMID- 12627193 TI - Further statistics in dentistry. Part 10: Sherlock Holmes, evidence and evidence based dentistry. PMID- 12627194 TI - Ecstasy related periodontitis and mucosal ulceration -- a case report. AB - Methylenedioxymethamphetamine (MDMA) more commonly known as 'Ecstasy' is a widely used recreational drug. The oral and systemic effects associated with its use have been well documented. This paper highlights a previously unreported complication of MDMA use on the oral mucosa. MDMA periodontitis is illustrated with a case report and the local oral and systemic effects of MDMA use outlined. PMID- 12627199 TI - An in vitro evaluation of the availability of cetylpyridinium chloride and chlorhexidine in some commercially available mouthrinse products. AB - OBJECTIVES: To assess the comparative activity of mouthrinses containing cetylpyridinium chloride (CPC) and chlorhexidine using the propensity to cause extrinsic staining in vitro as the outcome variable. METHODS: Saliva-coated clear acrylic specimens were exposed to cyclical staining regimens of either CPC or chlorhexidine followed by tea. Water and 0.2% chlorhexidine were used as negative and positive controls respectively. Staining cycles were repeated until an optical density of > 2 was reached by one of the products. RESULTS: For CPC there was a highly significant difference in staining between the products. Two CPC products performed numerically little better than water. For the chlorhexidine products the 0.2% formulation produced the most staining although little more than the UK version of the 0.1% rinse. The French 0.1% rinse produced by the same manufacturer as the UK formulation showed markedly reduced staining potential although significantly greater than water. CONCLUSION: This study, supported by previous in vitro and in vivo studies, indicates discrepancies in the availability of CPC and chlorhexidine in some mouthrinse products. Importantly, this may have an effect on the potential of some rinses to provide the expected plaque inhibitory activity. PMID- 12627200 TI - Retrospective long term monitoring of tooth wear using study models. AB - OBJECTIVE: Tooth wear is recognised as a common feature of European dentitions. However, little is known about its progression in susceptible patients. The aim of this study was to assess the degree and progression of tooth wear in patients by examining study casts taken of their teeth on two separate occasions. DESIGN: Over 500 sets of study casts taken during an 18-year period from patients referred for a variety of restorative procedures, were examined at Guy's Dental Hospital. Of these, 34 cases were found to have consecutive models taken at two time intervals and these were used to assess the progression of tooth wear. Study models from 19 females and 16 males, with an average age of 26 years (range 18 60) at the time of their first presentation and were all examined by a single operator. The Smith and Knight tooth wear index was used to assess the degree of tooth wear at presentation and then at another time which was a median of 26 months (interquartile range 14 - 50 months) later. RESULTS: The most common initial TWI score per surface was 1, with 54% of surfaces affected at the first assessment and 57% at the second. Score 2 was less common (14% at both assessments) and the scores for 3 and 4 combined were relatively uncommon with 5% of surfaces effected. Minimal progression of tooth wear was observed on study casts with only 7.3% of surfaces involved. CONCLUSION: In this sample, tooth wear was a slow, minimally progressive process. PMID- 12627204 TI - Focus awards 2002. Strelley Dental Clinic -- finalists for two years in a row. PMID- 12627201 TI - Evaluation of an oral health scoring system by dentists in general dental practice. AB - An Oral Health Index (OHX) has been designed to provide a numerical measure of the overall state of a patient's oral health by means of a series of simple clinical examinations. This has been amended to produce the Oral Health Score (OHS). OBJECTIVE: To assess, by means of a questionnaire, the ease of use and understanding of the OHS by general dental practitioners. METHODS: 350 GDPs were asked to participate in the project, of whom 329 agreed. These dentists were given a lecture of 2 hours' duration on the OHS. The participating dentists were requested to use the OHS for a period of 1 year. At the end of this period, a questionnaire was delivered to them by post, with an explanatory letter and reply paid envelope. RESULTS: Completed, usable questionnaires were received from 239 GDPs, a 77% response rate. Ninety three percent of respondents considered the OHS instructions to be satisfactory. The respondents' views on the criteria on which the OHS component assessments are based indicated that over 90% of respondents agreed with the criteria for caries, adequacy of restorations, periodontal assessment, mucosal assessment and assessment of dentures. A majority of respondents agreed that the OHS provided a valid representation of oral health. CONCLUSION: The results of the present study indicate that the OHS is considered to be an easy-to-use measure of a patient's oral health and that it provides a valid representation of a patient's oral health. PMID- 12627205 TI - Transfer technology in dentistry. PMID- 12627209 TI - Avoidance and management of positive surgical margins before, during and after radical prostatectomy. AB - Positive surgical margins after radical prostatectomy lead to an increased risk of progression and reduced disease free survival. Earlier detection of prostate cancer, appropriate patient selection and improved operative techniques can reduce the incidence of positive margins, though the risk can not be eliminated as pre-operative staging techniques are not sufficiently sensitive. Nerve sparing and bladder neck sparing do not adversely affect margin status in appropriately selected men. Once positive margins have been diagnosed the optimal management and the timing of treatment remains controversial. Adjuvant radiotherapy or salvage radiotherapy in men with a low PSA may improve local control and PSA free survival in some individuals, a survival benefit has not yet been established. PMID- 12627211 TI - Accuracy and repeatability of prostate volume measurements by transrectal ultrasound. AB - We evaluated six alternative methods of prostate volume determination by transrectal ultrasound, three based on planimetry and three based on measurement of prostate diameters. Prostate volume measurements were made on an average of 6.5 occasions over a 3 y period on 41 patients with benign prostatic hyperplasia, using standard techniques. We defined the average of multiple planimetries as the prostate reference volume. Agreement with the reference volume and reproducibility at repeat testing was in the same range for single planimetry and volume determinations based on the formulas height (H) x width (W) x length (L) x pi/6 and W x W x H x pi/6, but was poorer using the formula W x W x W x pi/6. Using the average result of two successive planimetry measurements increased the reproducibility of planimetry, being statistically significantly better than for one single planimetry (P=0.024) or for the formula W x W x H x pi/6 (P=0.048). Our study suggests that the simple formula based methods of prostate volume determination provide results that are only marginally inferior to one single planimetry, but results are improved by performing two planimetry measurements. PMID- 12627210 TI - The potential role of bisphosphonates in prostate cancer. AB - Skeletal morbidity secondary to metastases and osteoporosis is common in patients with advanced prostate cancer. Despite the typically sclerotic nature of prostate cancer metastases, osteoclast mediated osteolysis may play a significant role. This review addresses the newly recognised antitumour effects of bisphosphonates in addition to their role in inhibiting osteoclast mediated bone resorption. Both preclinical and clinical evidence of a role for bisphosphonates in the treatment and prevention of bone metastases secondary to prostate cancer is assessed. PMID- 12627212 TI - Pathological and clinical characteristics of large prostate cancers predominantly located in the transition zone. AB - Prostate carcinomas located in the transition zone are suspected to behave differently from the more frequent peripheral zone cancers. In this study, large transition zone prostate cancers were investigated for pathological and clinical features. From 365 consecutive radical prostatectomy specimens, 73 cases were disclosed with tumours larger than 10 cm(3). Of these, 14 were predominantly (>70% tumour area) located in the transition zone. Pathological investigations included a complete histological work-up, immunohistochemistry for p53 and bcl-2, and interphase cytogenetics for chromosomes 7, 8, 17, and X. Despite large tumour volumes and high preoperative prostate specific antigen (PSA)-values, most tumours showed quite favourable pathological features. Only two of these patients suffered from a postoperative PSA-recurrence during a median follow-up of 50 months. For comparison, 36 cases that contained tumours predominantly located in the peripheral zone mostly displayed adverse prognostic signs and 68.8% of these patients suffered from postoperative PSA-recurrence. We conclude that the peculiar pathological and clinical characteristics of large prostate cancers in the transition zone might be important for prognostic considerations. PMID- 12627214 TI - Urethral catheter removal 3 days after radical retropubic prostatectomy is feasible and desirable. AB - The purpose of this work was to assess the feasibility of urethral catheter removal 3 days after radical retropubic prostatectomy (RRP). Twenty-two patients who underwent RRP with a watertight eight-suture vesicourethral anastomosis had their urethral catheter removed usually on postoperative day (POD) 3. The average day of urethral catheter removal was POD 3.2. At 3 months, 56% of patients required no or one protective pad to stay dry and 68.4% of patients 'never leaked' or 'leaked occasionally'. Following RRP, the urethral catheter can be removed as early as POD 3 if the intraoperative anastomosis is watertight without compromising urinary continence. PMID- 12627213 TI - Outcome and complications of radical prostatectomy in patients with PSA <10 ng/ml: comparison between the retropubic, perineal and laparoscopic approach. AB - The objective of this work was to evaluate the oncological outcomes and complications of prostate cancer patients with prostate specific antigen (PSA)<10 ng/ml after radical prostatectomy by retropubic, perineal and laparoscopic approach. From 1988 to 2001, 306 patients with PSA<10 ng/ml underwent radical prostatectomy by the retropubic, perineal or laparoscopic approach. Mean operative time, complication rates, length of hospital stay, catheterization time and pathological results were reviewed. Kaplan-Meier analysis was used to evaluate the likelihood of biochemical PSA recurrence. There were no statistical differences between the three groups in terms of preoperative characteristics except for PSA levels (5.5, 6.5 and 6.6 ng/ml for the retropubic, perineal, and laparoscopic approach, respectively, P<0.05) and for the T1c stage prevalence (50%, 43.1% and 68.4%, P<0.05). Operating time was significatively longer in the laparoscopic approach (266 min), whereas transfusion rate (22.1%), bladder catetherization (12.1 days), and length of hospital stay (12.1 days) were higher in the retropubic group (P<0.05). The percentage of medical and surgical complications were 6.9%, 3.1% and 3.4% (P<0.05) and 18.6%, 16.9% and 11.6% (P<0.05) for the retropubic, perineal, and laparoscopic approach, respectively. Pathological staging revealed pT2 in 76.7%, 78.4% and 81.3% for retropubic, perineal and laparoscopic approach, respectively (P<0.05). Positive surgical positive margins were noted in 20.9%, 18.4% and 20.6% (P>0.05). The actuarial 3 year recurrence-free survival rate was 89.3%, 89.2% and 86.2% (P>0.05) for retropubic, perineal and laparoscopic approach, respectively. It can be concluded that in patients with preoperative PSA<10 ng/ml, clinical outcome and complication rates were similar, regardless of the choice of surgical approach. PMID- 12627215 TI - Evolution of toxicity after conformal radiotherapy for prostate cancer. AB - The limiting factor for radiation (RT) dose-escalation is normal tissue toxicity. In dose-escalation studies, it is important to determine the factors associated with toxicity and the length of follow-up period after which a particular RT dose is considered safe. We analyzed 449 prostate cancer patients treated with RT at our institution and followed for a median of 27 months. Genitourinary (GU) and gastrointerological (GI) complications were graded and analyzed using three different statistical models. Univariate and multivariate analyses were conducted for factors associated with toxicity. There was no RTOG grade 4 or 5 toxicity. Only 23 patients (5%) experienced grade 3 toxicity. After treatment, there was an initial rapid decline in the risk of toxicity following treatment, followed by an increase or stabilization of the toxicity with time of follow-up. The breakpoints between the two periods were 2 y (any toxicity) and 1 y (high toxicity) for GU and 9 months (any toxicity, high toxicity) for GI. Age, dose, fraction size, duration of treatment and hospital of treatment emerge as important factors in the probability of developing toxicity. Our study shows that delivering conventional doses using conformal techniques is associated with minimal high grade toxicity. However, even within a narrow dose range and fraction size used, differences do emerge which should lead one to be cautious in extending the results of dose escalation study to the community practice without a sufficient follow-up. PMID- 12627216 TI - Androgen deprivation in men with prostate cancer is associated with an increased rate of bone loss. AB - The objective of this work was to determine the effect of androgen deprivation therapy (ADT) on rates of bone mineral density (BMD) loss in men with prostate cancer. It was a prospective study comparing men receiving ADT to age matched controls for 2 y. Subjects received a history, physical exam, bone mineral density measurement, and laboratory evaluation every 6 months. Thirty-nine subjects receiving continuous ADT for prostate cancer (subjects) were compared to 39 age-matched controls not receiving ADT (controls). Twenty-three subjects and 30 controls completed the study through 24 months. Men in the ADT group demonstrated greater rates of bone mineral density loss than men in the control group at every site except the lumbar spine. Twenty-four month per cent of bone mineral density loss is presented as mean+/-standard error (s.e.). At the distal forearm, the ADT group value was -9.4%+/-1.0% and -4.4%+/-0.3% for controls (P<0.0005). The ADT group femoral neck values were -1.9%+/-0.7% and 0.6%+/-0.5% in the control group (P=0.0016). The ADT group total hip value was -1.5%+/-1.0% and 0.8%+/-0.5% in the control group (P=0.0018). The ADT group trochanter value was -2.0%+/-1.3% and -0.1%+/-0.5% in the control group (P=0.0019). The ADT group lumbar spine value was -0.2%+/-0.8 % and 1.1%+/-0.6% in the control group (P=0.079). Our data demonstrate greater rates of bone mineral density loss in men receiving androgen deprivation therapy for prostate cancer. PMID- 12627217 TI - A Nod Scid mouse model to study human prostate cancer. AB - Prostate cancer is the second cause of cancer mortality in men in Western countries. To study new therapeutic approaches such as gene therapy, animal models of human prostate cancer with metastatic behavior are mandatory. We used the Nod Scid mouse strain to develop an orthotopic animal model. Two androgen independent cell lines (PC-3 and DU 145) were used. Local tumor growth and metastases were analyzed. The tumor take rates were close to those reported in the literature. However, a high frequency of various metastatic sites has been observed (liver, lung, spleen, adrenal, kidney, lymph node, and diaphragm). It can be concluded that the Nod Scid mouse is a relevant preclinical animal model to study human prostate cancer. Metastatic sites seem more numerous in comparison to other orthotopic mice models described. PMID- 12627218 TI - Gene therapy for prostate cancer: toxicological profile of four HSV-tk transducing adenoviral vectors regulated by different promoters. AB - Adenoviral vector delivery of the Herpes simplex virus thymidine kinase (HSV-tk) gene in combination with the prodrug ganciclovir (GCV) has been tested in phase I clinical trials for prostate cancer and found to exhibit a satisfactory toxicity profile. We have developed additional adenoviral vectors with differing promoters to optimize the expression profile and in the present study evaluate the potential systemic toxicity of these vectors. Four recombinant adenoviral vectors that express the HSV-tk gene were generated using three different promoters: CMV (leftward orientation); RSV (both rightward and leftward orientation); and the mouse caveolin-1 (cav-1) promoter (leftward orientation). Efficacy was determined in vitro by cytotoxicity assays in a mouse prostate cancer cell line, RM-9, and in vivo by treating orthotopic tumors. Potential toxicity was evaluated from liver histology and apoptotic cell counts and enzyme levels in the serum following intravenous adenoviral vector injection. Although there were differences in HSV-tk expression at the protein level among the four vectors there were no significant differences in in-vitro cytotoxicity studies with GCV or in vivo in tumor growth suppression of an orthotopic mouse prostate cancer model in GCV treated mice. Intravenous delivery of high doses of all adenoviral vectors lead to abnormalities in liver function as measured by specific serum markers and histological evaluation of liver tissue and increased levels of apoptosis in the liver. These abnormalities were most prevalent with the vector containing the CMV promoter and the rightward oriented RSV promoter. They were least prevalent in the vector regulated by the cav-1 promoter. Upregulation of specific chemokines, MIP-2 and MIP-1beta was correlated with apoptotic counts. Our results demonstrate that comprehensive toxicological analysis of adenoviral vectors provides internally consistent information that can differentiate vectors with comparable efficacy based on toxicity. In these studies vectors with the cav 1 promoter-driven and leftward RSV-driven HSV-tk gene demonstrated minimal toxicities with cytotoxic effectiveness comparable to more toxic vectors. Our studies further suggest that promoter selection can influence the toxic effects of an adenoviral gene therapy vector. PMID- 12627223 TI - Mechanism of metal activation of human hematopoietic prostaglandin D synthase. AB - Here we report the crystal structures of human hematopoietic prostaglandin (PG) D synthase bound to glutathione (GSH) and Ca2+ or Mg2+. Using GSH as a cofactor, prostaglandin D synthase catalyzes the isomerization of PGH2 to PGD2, a mediator for allergy response. The enzyme is a homodimer, and Ca2+ or Mg2+ increases its activity to approximately 150% of the basal level, with half maximum effective concentrations of 400 microM for Ca2+ and 50 microM for Mg2+. In the Mg2+-bound form, the ion is octahedrally coordinated by six water molecules at the dimer interface. The water molecules are surrounded by pairs of Asp93, Asp96 and Asp97 from each subunit. Ca(2+) is coordinated by five water molecules and an Asp96 from one subunit. The Asp96 residue in the Ca2+-bound form makes hydrogen bonds with two guanidium nitrogen atoms of Arg14 in the GSH-binding pocket. Mg2+ alters the coordinating water structure and reduces one hydrogen bond between Asp96 and Arg14, thereby changing the interaction between Arg14 and GSH. This effect explains a four-fold reduction in the K(m) of the enzyme for GSH. The structure provides insights into how Ca2+ or Mg2+ binding activates human hematopoietic PGD synthase. PMID- 12627222 TI - Structural insights into the U-box, a domain associated with multi ubiquitination. AB - The structure of the U-box in the essential Saccharomyces cerevisiae pre-mRNA splicing factor Prp19p has been determined by NMR. The conserved zinc-binding sites supporting the cross-brace arrangement in RING-finger domains are replaced by hydrogen-bonding networks in the U-box. These hydrogen-bonding networks are necessary for the structural stabilization and activity of the U-box. A conservative Val-->Ile point mutation in the Prp19p U-box domain leads to pre mRNA splicing defects in vivo. NMR analysis of this mutant shows that the substitution disrupts structural integrity of the U-box domain. Furthermore, comparison of the Prp19p U-box domain with known RING-E2 complex structures demonstrates that both U-box and RING-fingers contain a conserved interaction surface. Mutagenesis of residues at this interface, while not perturbing the structure of the U-box, abrogates Prp19p function in vivo. These comparative structural and functional analyses imply that the U-box and its associated ubiquitin ligase activity are critical for Prp19p function in vivo. PMID- 12627224 TI - The nature of the di-iron site in the bacterioferritin from Desulfovibrio desulfuricans. AB - The first crystal structure of a native di-iron center in an iron-storage protein (bacterio)ferritin is reported. The protein, isolated from the anaerobic bacterium Desulfovibrio desulfuricans, has the unique property of having Fe coproporphyrin III as its heme cofactor. The three-dimensional structure of this bacterioferritin was determined in three distinct catalytic/redox states by X-ray crystallography (at 1.95, 2.05 and 2.35 A resolution), corresponding to different intermediates of the di-iron ferroxidase site. Conformational changes associated with these intermediates support the idea of a route for iron entry into the protein shell through a pore that passes through the di-iron center. Molecular surface and electrostatic potential calculations also suggest the presence of another ion channel, distant from the channels at the three- and four-fold axes proposed as points of entry for the iron atoms. PMID- 12627225 TI - Ni-Zn-[Fe4-S4] and Ni-Ni-[Fe4-S4] clusters in closed and open subunits of acetyl CoA synthase/carbon monoxide dehydrogenase. AB - The crystal structure of the tetrameric alpha2beta2 acetyl-coenzyme A synthase/carbon monoxide dehydrogenase from Moorella thermoacetica has been solved at 1.9 A resolution. Surprisingly, the two alpha subunits display different (open and closed) conformations. Furthermore, X-ray data collected from crystals near the absorption edges of several metal ions indicate that the closed form contains one Zn and one Ni at its active site metal cluster (A-cluster) in the alpha subunit, whereas the open form has two Ni ions at the corresponding positions. Alternative metal contents at the active site have been observed in a recent structure of the same protein in which A-clusters contained one Cu and one Ni, and in reconstitution studies of a recombinant apo form of a related acetyl CoA synthase. On the basis of our observations along with previously reported data, we postulate that only the A-clusters containing two Ni ions are catalytically active. PMID- 12627226 TI - Identification and isolation of multipotential neural progenitor cells from the subcortical white matter of the adult human brain. AB - The subcortical white matter of the adult human brain harbors a pool of glial progenitor cells. These cells can be isolated by fluorescence-activated cell sorting (FACS) after either transfection with green fluorescent protein (GFP) under the control of the CNP2 promoter, or A2B5-targeted immunotagging. Although these cells give rise largely to oligodendrocytes, in low-density culture we observed that some also generated neurons. We thus asked whether these nominally glial progenitors might include multipotential progenitor cells capable of neurogenesis. We found that adult human white-matter progenitor cells (WMPCs) could be passaged as neurospheres in vitro and that these cells generated functionally competent neurons and glia both in vitro and after xenograft to the fetal rat brain. WMPCs were able to produce neurons after their initial isolation and did not require in vitro expansion or reprogramming to do so. These experiments indicate that an abundant pool of mitotically competent neurogenic progenitor cells resides in the adult human white matter. PMID- 12627227 TI - Geographic diversity and temporal trends of antimicrobial resistance in Streptococcus pneumoniae in the United States. AB - Resistance of Streptococcus pneumoniae to antibiotics is increasing throughout the United States, with substantial variation among geographic regions. We show that patterns of geographic variation are best explained by the intensity of selection for resistance, which is reflected by differences between the proportions of resistance within individual serotypes, rather than by differences between the frequencies of particular serotypes. Using a mathematical transmission model, we analyzed temporal trends in the proportions of singly and dually resistant organisms and found that pneumococcal strains resistant to both penicillin and erythromycin are increasing faster than strains singly resistant to either. Using the model, we predict that by 1 July 2004, in the absence of a vaccine, 41% of pneumococci at the Centers for Disease Control and Prevention (CDC)'s Active Bacterial Core surveillance (ABCs) sites, taken together, will be dually resistant, with 5% resistant to penicillin only and 5% to erythromycin only. PMID- 12627228 TI - Grey-lethal mutation induces severe malignant autosomal recessive osteopetrosis in mouse and human. AB - The spontaneous mouse grey-lethal (gl) mutation is responsible for a coat color defect and for the development of the most severe autosomal recessive form of osteopetrosis. Using a positional cloning approach, we have mapped and isolated the gl locus from a approximately 1.5 cM genetic interval. The gl locus was identified in a bacterial artificial chromosome (BAC) contig by functional genetic complementation in transgenic mice. Genomic sequence analysis revealed that the gl mutation is a deletion resulting in complete loss of function. The unique approximately 3 kb wild-type transcript is expressed primarily in osteoclasts and melanocytes as well as in brain, kidney, thymus and spleen. The gl gene is predicted to encode a 338-amino acid type I transmembrane protein that localizes to the intracellular compartment. Mutation in the human GL gene leads to severe recessive osteopetrosis. Our studies show that mouse Gl protein function is absolutely required for osteoclast and melanocyte maturation and function. PMID- 12627229 TI - Hypergammaglobulinemia and autoantibody induction mechanisms in viral infections. AB - Polyclonal hypergammaglobulinemia is a characteristic of chronic inflammatory conditions, including persisting viral infections and autoimmune diseases. Here we have studied hypergammaglobulinemia in mice infected with lymphocytic choriomeningitis virus (LCMV), which induces nonspecific immunoglobulins as a result of switching natural IgM specificities to IgG. The process is dependent on help from CD4+ T cells that specifically recognize LCMV peptides presented by B cells on major histocompatibility complex class II molecules. Thus, hypergammaglobulinemia may arise when specific helper T cells recognize B cells that have processed viral antigens irrespective of the B cell receptor specificity. This nonspecific B cell activation may contribute to antibody mediated autoimmunity. PMID- 12627230 TI - Loss-of-function mutations in FGFR1 cause autosomal dominant Kallmann syndrome. AB - We took advantage of overlapping interstitial deletions at chromosome 8p11-p12 in two individuals with contiguous gene syndromes and defined an interval of roughly 540 kb associated with a dominant form of Kallmann syndrome, KAL2. We establish here that loss-of-function mutations in FGFR1 underlie KAL2 whereas a gain-of function mutation in FGFR1 has been shown to cause a form of craniosynostosis. Moreover, we suggest that the KAL1 gene product, the extracellular matrix protein anosmin-1, is involved in FGF signaling and propose that the gender difference in anosmin-1 dosage (because KAL1 partially escapes X inactivation) explains the higher prevalence of the disease in males. PMID- 12627231 TI - Mutant dynactin in motor neuron disease. AB - Impaired axonal transport in motor neurons has been proposed as a mechanism for neuronal degeneration in motor neuron disease. Here we show linkage of a lower motor neuron disease to a region of 4 Mb at chromosome 2p13. Mutation analysis of a gene in this interval that encodes the largest subunit of the axonal transport protein dynactin showed a single base-pair change resulting in an amino-acid substitution that is predicted to distort the folding of dynactin's microtubule binding domain. Binding assays show decreased binding of the mutant protein to microtubules. Our results show that dysfunction of dynactin-mediated transport can lead to human motor neuron disease. PMID- 12627232 TI - In vivo characterization of regulatory polymorphisms by allele-specific quantification of RNA polymerase loading. AB - In vivo characterization of regulatory polymorphisms is a key requirement for next-generation human genetic analysis. Here we describe haploChIP, a method that uses chromatin immunoprecipitation (ChIP) and mass spectrometry to identify differential protein-DNA binding in vivo associated with allelic variants of a gene. We demonstrate this approach with the imprinted gene SNRPN. HaploChIP showed close correlation between the level of bound phosphorylated RNA polymerase II at the SNRPN locus and allele-specific expression. Application of the approach to the TNF/LTA locus identified functionally important haplotypes that correlate with allele-specific transcription of LTA. The haploChIP method may be useful in high-throughput screening for common DNA polymorphisms that affect gene regulation in vivo. PMID- 12627233 TI - Genome imprinting regulated by the mouse Polycomb group protein Eed. AB - Epigenetic regulation is essential for temporal, tissue-specific and parent-of origin-dependent gene expression. It has recently been found that the mouse Polycomb group (PcG) gene Eed (embryonic ectoderm development) acts to maintain repression of the imprinted X chromosome. Here, we investigated whether Eed is also required for regulation of autosomal imprinted loci. Expression analyses showed that transcripts from the silent alleles of a subset of paternally repressed genes were present in Eed(-/-) embryos. Parent-of-origin methylation was preserved in these embryos, but we observed changes in the methylation status of specific CpGs in differentially methylated regions (DMRs) at affected but not at unaffected loci. These data identify Eed as a member of a new class of trans acting factors that regulate parent-of-origin expression at imprinted loci. PMID- 12627234 TI - [Diagnosis related groups in psychiatry and psychotherapy--alternatives to DRGs]. AB - Diagnosis related groups (DRGs) will be introduced in Germany in 2003 as a basis for hospital funding in all areas except for psychiatry and psychotherapeutic medicine. The following article provides an overview of the use of alternative casemix-systems in the treatment of patients with mental disorders. Overall, DRGs and alternative systems have proven to be poor predictors of resource consumption due to the high variation in the length of stay and the variable treatment opportunities. Therefore, a DRG-system must include the possibility of coded therapeutic treatments, analogous to the procedures used in surgical DRGs, in order to integrate psychiatry and psychotherapeutic medicine into a DRG-based payment system. PMID- 12627235 TI - [Allocation of cognitive resources during the simultaneous performance of cognitive and sensorimotor tasks]. AB - We review research on the allocation of cognitive resources during the simultaneous performance of cognitive and sensorimotor tasks. From the developmental and clinical perspectives,we emphasize: (1) the distinction between the availability and the allocation of resources, (2) lifespan changes in relation to the environmental validity of sensorimotor functions, and (3) the potentials and limitations for an individuals' adaptations to multi-task constraints. These aspects can be operationalized within the framework of selection, optimization, and compensation (SOC). Related studies focus on older individuals' selective resource allocation and compensatory processes as adaptive means in the context of reduced resources and decreased sensorimotor functioning. Results show that older adults must invest increasing amounts of their cognitive resources into the coordination of bodily functions such as balance and gait. SOC research on Alzheimer's patients provides new insights into the increased risks of falling. We argue that adaptive resource allocation in everyday sensorimotor performance is an instance of intelligent behavior that is insufficiently represented in extant psychometric tests. PMID- 12627236 TI - [Outpatient alcohol detoxification]. AB - Increasing significance is being attributed to outpatient detoxification treatment of alcohol-dependent patients. Many patients want to undergo outpatient detoxification or even carry it out without professional supervision. In some Scandinavian countries and in the USA, outpatient detoxification under medical supervision has increased substantially. In these countries,numerous studies describe outpatient alcohol detoxification as a safe and cost-reducing method. These studies vary in many aspects, especially regarding the nature and dose of the withdrawal medication, but also concerning the inclusion criteria of patients. Medications to treat or prevent alcohol withdrawal symptoms are rather heterogeneous. Besides well-known standard medication established in inpatient treatment (e.g.,benzodiazepines and carbamazepine), interesting studies report the application of rather uncommon substances such as physostigmine and psychotropic analgesic nitrous oxide (PAN) for alcohol detoxification. This article provides an overview on outpatient alcohol detoxification and discusses its transfer to practical application. PMID- 12627237 TI - [New possibilities in treatment and rehabilitation of alcohol-dependent patients- a catamnestic study on the efficiency of outpatient treatment programmes demonstrated by a model procedure]. AB - First results of a clinical and catamnestic investigation are reported for the efficiency of a highly structured outpatient therapy with alcohol-dependents. One hundred and two patients were included in the study. Of the patients,60% were male and 40% female. The average age was 45 years (+/-8). The average duration of alcohol dependence amounted to 15 years (+/-9), and the last average quantity of pure alcohol drunk was 193 g. Twenty-seven per cent of the patients had completed inpatient therapies in the past. Treatment retention amounted to n=74 (72.5%), and 18 of the 25 dropped out because of alcohol relapse. On average, relapsed dropouts indicated a longer abuse of alcohol and significantly more pretreatments than completers, and they also reported significantly stronger craving for alcohol (measured with the OCDS). Furthermore, they also achieved significantly higher total scores in the BDI (depression) and STAI (anxiety) scales at the beginning of therapy. At 6/12-month follow-ups, 90%-95% of the patients were successfully located and interviewed. Analyses revealed that 64% of the patients were still abstinent at 6-month follow-up evaluation, and 56% had remained abstinent until 12-month follow-up. Therapeutic implications of these satisfying therapy results are discussed, and the current knowledge on the efficiency of outpatient therapies is presented. PMID- 12627238 TI - [Standardized documentation procedure as a basis for improvement of process quality of treatment in psychiatric hospitals]. AB - A standardized documentation system is presented for detailed description and analysis of treatment procedure for psychiatric inpatients. In the first step of a pilot study limited to patients with depressive syndromes at the Department of Psychiatry, University of Mainz, for each subject the clinical status was assessed weekly using psychopathological ratings and depicted in its temporal course along with therapeutic activities. Visualization of the treatment process turned out to be useful for everyday work by supporting the application of practice guidelines for clinical decisions. Beyond optimization of the individual treatment process, aggregation of the data also allows global assessment of treatment processes in the clinic. Thus, in addition to established basic documentation, the presented documentation procedure allows the development of a powerful quality management system in psychiatric clinics, due to its process orientation. Moreover, in psychiatric care research, it is an adequate basis for evaluating the effectiveness and efficiency of treatment procedures. PMID- 12627239 TI - [The 4-week prevalence of somatoform disorders and associated psychosocial impairment]. AB - In the course of a WHO study,we report on the prevalence of somatoform disorders (SFD) and the associated psychosocial impairment in five western German primary care settings. In accordance with ICD-10 classification, a 4-week prevalence of 28.5% was found for SFD (number of patients in the age between 18 and 60 with an SFD in the last 28 days). The accumulation of SFD was higher in female patients than in males (RR 1.7), in particular when the number of children was >1 (RR 1.8). The female-male difference was more marked in persistent somatoform pain disorder (RR 2.1) and unspecific somatization disorder (RR 5.0). Concerning other psychiatric disorders, neurasthenia occurred most frequently,with a 4-week prevalence of 8.2%. The 4-week prevalence of concomitant occurrence of SFD and other psychiatric disorders was 7.7%. Working capability was most severely impaired, with 22.5 days of absence from work during the last month, in male patients with hypochondriacal disorder. In comparison, somatization disorder resulted in a severe level of psychosocial impairment, with 10.3 days of absence in work during the last month in female patients. The coexistence of SFS with other psychiatric disorders resulted in a greater extent of psychosocial impairment. PMID- 12627240 TI - [On the admittance of forensic patients to general psychiatric wards]. AB - In Germany, due to an increasing number of "hospital order sentences," the capacities of forensic hospitals are exhausted. In the late 1990s,general psychiatric hospitals admitted a remarkable number of mentally disturbed offenders. In this study,data of 140 patients treated in general psychiatric hospitals in the German lower Rhine region are presented. These patients clearly differ from those treated in forensic hospitals. Approximately 60% of them suffer from schizophrenic psychosis. Only a few reveal a long antisocial background, a severe personality disorder, or a sexual offence as an index crime. Though the staff in the general psychiatric units often complain about the circumstances of patients' admittance to the hospitals, in about half of the cases they do not agree with a transferring of their patients to a forensic hospital. The results of this study are discussed in regard to general questions of organizing forensic psychiatric treatment. PMID- 12627241 TI - [A risk profile of the aggressive psychiatric inpatient: can it be identified?]. AB - Violent behavior of psychiatric inpatients has grown in interest. Within the scope of a prospective study,patient assaults in six psychiatric hospitals of the Swiss Canton of Zurich were assessed during a period of 6 months, from November 1999 until May 2000. Three hundred eighty-eight of 5,251 inpatients (7.4%) were reported to act aggressively. The following risk factors for aggressive behavior were identified: male gender, younger age, unemployment, severity of mental illness, and rehospitalization. Psychiatric diagnosis was not related to aggressiveness. In spite of the detection of risk factors, a characteristic profile of aggressive psychiatric inpatients cannot be stated. Research should pay more attention to situation-specific cues for the aggressive behavior of such individuals. PMID- 12627242 TI - [First onset of panic disorder and agoraphobia induced by a series of inappropriate shocks of an implanted cardioverter/defibrillator]. AB - We report on a 61-year-old patient with an implanted defibrillator due to malignant arrhythmia after myocardial infarction. After several years of appropriate function, the patient suffered a cluster of 16 shocks due to a defect in the defibrillator. Soon the patient developed a panic disorder with agoraphobia which was successfully treated with cognitive-behavioral therapy. Possible cognitive-behavioral mechanisms of panic disorder and special issues of behavioral therapy of patients with serious organic diseases are discussed. PMID- 12627243 TI - [Self-assessment of post-traumatic stress reactions in the wake of the September 11, 2001, terrorist attacks in New York City. A survey among medical students]. AB - The present paper describes psychological sequelae of the September 11 terrorist attacks in New York City in a cohort of 174 medical students (104 females, 70 men, age 18 to 37 years) in their first academic year at the University of Kiel,Germany. For self-report of traumatic stress reactions,the Revised Impact of Event Scale (IES-R) was administered 6 and 9 weeks after the disaster. The students reported weak to moderate levels of distress, the average IES-R scores on the intrusion and avoidance subscales were 11.1 (SD 6.2) and 10.6 (SD 6.4), respectively, and 5.2 (SD 4.1) for the hyperarousal scale. All symptoms had clearly faded at 9 weeks. Students with traumatic life events indicated significantly higher scores in some avoidance items. The current data suggest that the IES-R can be used as a screening measure in future research of trauma related stress reactions also in people exposed to catastrophes by media coverage. PMID- 12627245 TI - [The neuroendocrinology of stress and the pathophysiology and therapy of depression and anxiety]. AB - Clinical and preclinical studies have gathered substantial evidence that stress response alterations play a major role in the development of major depression, panic disorder, and post-traumatic stress disorder. The stress response, the hypothalamic pituitary adrenocortical (HPA) system and its modulation by corticotropin-releasing hormones (CRH),corticosteroids,and their receptors, and the roles of natriuretic peptides and neuroactive steroids are described. We review the role of the HPA system in major depression, panic disorder, and post traumatic stress disorder and its possible relevance for treatment. Impaired glucocorticoid receptor function in major depression is associated with an excessive release of neurohormones such as CRH, to which a number of signs and symptoms characteristic of depression can be ascribed. In panic disorder, a role of central CRH in panic attacks has been suggested. Atrial natriuretic peptide (ANP) is causally involved in sodium lactate-induced panic attacks. Furthermore, preclinical and clinical data on its anxiolytic activity suggest that nonpeptidergic ANP receptor ligands may be potentially useful in the treatment of anxiety disorders. Post-traumatic stress disorder is characterized by a peripheral hyporesponsive HPA system and elevated CRH concentrations in the CSF. This dissociation is probably related to an increased risk of this disorder. We further review recent data that describe an important role of GABA(A)-receptor modulatory,3 alpha-reduced neuroactive steroids in major depression, anxiety, and its treatment. Antidepressants are effective in both depression and anxiety disorders and have major effects on the HPA system,especially on glucocorticoid and mineralocorticoid receptors. Normalization of HPA system abnormalities is a strong predictor of the clinical course, at least in major depression and panic disorder. Currently,CRH-R1 or glucocorticoid receptor antagonists and ANP receptor agonists are being studied and may provide future treatment options more closely related to the pathophysiology of these disorders. PMID- 12627244 TI - [Neurobiology and genetics of anxiety in an animal model]. AB - This paper presents a valid animal model of innate anxiety/depression: anxious (HAB) or non-anxious (LAB) rats, which show stable and robust responses in a variety of ethological tests. In addition to their extreme anxiety-related behavior, HAB animals are characterized by passive stress coping, an activated stress (HPA) axis, and increased stress vulnerability. The enhanced expression and release of arginine vasopressin (AVP) in the hypothalamus of HAB rats seem to underlie these phenomena. Accordingly, an AVP receptor antagonist attenuates anxiety-related behavior and normalizes the HPA axis and the dexamethasone/CRH test. Treatment with the antidepressive drug paroxetine reduces the overexpression of AVP and normalizes both the depression-like behavior and neuroendocrine correlates of anxiety/depression. The complex phenotyping led us to the conclusion that the AVP gene is likely to be a candidate gene of inborn anxiety. Partial genotyping of HAB animals results in the identification of polymorphisms (SNPs) in the promoter domain of the AVP gene, thus potentially leading to novel strategies of diagnosis and therapy of anxiety disorders and depression. PMID- 12627246 TI - [Cancer from snuff?]. PMID- 12627247 TI - [Diagnosis and therapy of syndromic and non-syndromic craniosynostosis]. AB - Premature craniosynostosis is caused by a premature closure of single or multiple sutures of the cranial vault. It may result not only in aesthetic but also in functional disorders with an occasional ICP increase leading to a dramatic loss of vision. In the 1970s,Paul Tessier published his craniofacial principles for the primary treatment of craniosynostosis. His approach has gained wide acceptance over the previously described craniectomies and bone flaps, especially because of the significant benefits with an acceptably low level of morbidity. The following paper reviews the different kinds of craniosynostoses, their specific clinical manifestations, the necessary diagnostic examinations and the specific surgical procedures and operative results. PMID- 12627248 TI - [The application of an optical navigation system in endonasal sinus surgery]. AB - BACKGROUND: There are several CAS-systems in use in modern endonasal sinus surgery. The VectorVision Compact is the first optoelectrical system especially designed for such surgery. This paper investigates its reliability and practicability. PATIENTS AND METHODS: The system was used for intraoperative navigation in 60 patients suffering from chronic rhinosinusitis. The accuracy was measured by visual estimation and compared with other navigated systems. RESULTS: The accuracy was 1.9 mm which is comparable to other systems on the market. Navigation was successful and reliable in 83.3% of the patients. Problems occurred mainly due to loss of sight of the referencing balls and when using other surgical instruments. CONCLUSION: The VectorVision Compact was adequate for endonasal sinus surgery. Further improvements incorporating various surgical instruments should broaden its clinical usefulness. PMID- 12627249 TI - [Surgical therapy of Madelung's disease in the head and neck area]. AB - PATIENTS AND METHODS: Between 1995 and 2001 we treated 12 patients with benign symmetric lipomatosis in the head and neck. The group comprised 11 men and 1 woman aged 34-62 (M=47). The most frequent complaints were diminished range of motion of the head and obstruction when eating or speaking. Combined lipectomy and liposuction were performed in all patients, with liposuction being carried out in a second session. RESULTS: The mean follow-up period was 3 years. The functional results were very satisfactory in all patients. Of the 12 patients, 9 were also satisfied with the aesthetic outcome.A recurrence occurred in three patients 1.5, 2, or 3 years after the operation. Severe complications were not observed. CONCLUSIONS: According to our experience, combined lipectomy and liposuction represents a successful procedure in treating benign symmetric lipomatosis in the head and neck region.Nevertheless, advantages and drawbacks of the two techniques should be equally considered prior to surgery. PMID- 12627250 TI - [Bilateral vestibular loss. Diagnosis and follow-up]. AB - BACKGROUND: Atypical symptom in patients with bilateral vestibular loss is head movement-induced oscillopsia. The paucity of precise complaints in many patients is surprising. Therefore, bilateral loss of vestibular function is often undiagnosed. PATIENTS: We report on the long-term follow-up in 29 patients. They were monitored for 2-7 years (mean: 4.5 years). RESULTS: Of the 29 patients 16 described oscillopsia. All symptomatic patients had acute bilateral vestibular loss.Patients described that their symptoms improved over a period of 1-2 years. Improvement was not age dependent. CONCLUSIONS: Otoneurologists should be aware of the particular clinical symptoms in bilateral vestibular loss. Regarding clinical features, compensation of bilateral vestibular loss seems to be unlikely only based on central compensatory eye movement reflexes. More likely perceptual adaptations and restriction of head movement are responsible for subjective improvement. PMID- 12627251 TI - [How is tinnitus influenced by cochlear implantation?]. AB - INTRODUCTION: Tinnitus is one of the most common symptoms in patients with profound sensorineural hearing loss (SNHL). Cochlear implantation (CI), which has become a standard method to rehabilitate profound SNHL, has been also observed to effect tinnitus in many cases. In the Salzburg Cochlear Implant Center,more than 230 profoundly deaf patients, mainly children,have been provided with CI between 1992 and 2000. PATIENTS AND METHOD: In a retrospective study, 39 adult patients were assessed by questionnaire for the frequency of tinnitus, subjective loudness sensitivity, and tinnitus-specific impairment before and after the CI. RESULTS: Before implantation 26 patients had tinnitus, while 24 patients had the same symptoms after the CI. In 13 patients (50%) tinnitus remained unaffected by the operation. Intensity of tinnitus decreased in seven patients (26.7%), and in four patients (15.4%) it completely disappeared after the implantation.However, intensity increased in two patients (7.7%), and in another two patients, who had not experienced tinnitus before, it began first after the implantation. No correlation has been found between the daily using time of CI and the tinnitus impairment.Also no effect of tinnitus was seen on postoperative hearing assessed by Freiburg numbers, monosyllables, and Innsbruck sentence test scores. CONCLUSION: CI significantly reduces the subjectively perceived impairment, although the reduction in the subjectively perceived loudness of the tinnitus is not significant. PMID- 12627252 TI - [Advanced transfacial approach to extensive sellar adenoma of the pituitary gland]. AB - The excision of tumors of the pituitary gland is usually achieved via a transsphenoidal transnasal approach. In the past, alternative approaches were described for macroadenomas. Especially in tumors surrounding the internal carotid artery (ICA) and the optic nerve, the selection of the appropriate approach is of importance. We demonstrate the case of a second extensive relapse of a macroadenoma after primary transsphenoidal resection and radiation therapy. As the tumor enclosed both the ICA and left optic nerve, a median external approach via the upper midface was chosen. After the median skin incision and translocation of the upper parts of the nasal structure, the papyraceous lamina was removed bilaterally. The orbital tissue was laterally exposed with retractors, achieving a wide access to both carotids and optic nerves. The functional and aesthetic results were satisfactory. Compared with other approaches, this one provides a good exposure of both the ICA and optic nerve. PMID- 12627253 TI - [A bone-destroying tumor of the maxilla. Reparative giant cell granuloma or brown tumor?]. AB - Brown tumors are focal bone lesions caused by an increased osteoclastic activity and fibroblastic proliferation within a primary or more rarely secondary hyperparathyroidism. They are named after their typical brown hemorrhagic stroma with its also typical giant cell formations. We report the case of a 31-year-old pregnant patient with a rapidly growing tumor of her left maxilla whose first symptoms during pregnancy mimicked chronic sinusitis. After swelling of the cheek, diplopia, and recurrent epistaxis appeared, she was referred to our Department for further diagnostics. After CT scan, biopsy was performed under the presumption of a malignant process with the surprising histological result of a reparative giant cell granuloma. At the same time, hyperthyroidism and nodular goiter were diagnosed and further endocrinological examinations were planned. Not until a parathyroid adenoma was diagnosed after urgent operation of the maxillary process (loose molar teeth and displacement of the left bulbus) could the tumor be interpreted and detected within this context of primary hyperparathyroidism as a brown tumor. The brown tumor should be taken into consideration as a rare differential diagnosis of a bone-destroying process of the facial bones. We discuss the clinical signs, diagnostics, and therapy for this case as well as the relevant literature. The reparative giant cell granuloma represents an important differential diagnosis and cannot be distinguished from a brown tumor by histological examination or radiological findings without complete information about the clinical signs and the endocrinological status of the patient. PMID- 12627254 TI - [Recovery of normal auditory threshold after hearing damage from fireworks and signalling pistols]. PMID- 12627255 TI - [Haemostatic disorders in ENT patients. Part 2: Pathophysiology, diagnostics, clinical feature and therapy]. AB - In the first part of the medical education article on haemostatic disorders in ENT patients the basic physiology of haemostasis and main diagnostic tools were presented and discussed. The second part presents disorders of the coagulation system,thrombocyte function and blood vessels with special emphasis on clinical practice in ENT surgery. In this context, haemophilia A and B, von Willebrand disease and different forms of thrombocytopenia are of main clinical importance. Some underlying diseases such as malignomas, renal and hepatic insufficiency in combination with drug therapy (e.g.anticoagulants and thrombocyte function inhibitors) play an important role in clinical practice as well. Sepsis and haemorrhage may lead to disseminated intravascular coagulation (DIC). Beside a systematic review, important haemostatic disorders are illustrated with case reports. PMID- 12627256 TI - [Death inducing ligands in combination with ionizing radiation: objective and current knowledge]. AB - BACKGROUND: Apart from optimization of the radiation technology, future new strategies in radiation oncology will focus on the biological optimisation leading to improved adaptation of the tumor therapy on each tumor entity. In this regard, different areas of biological research may be distinguished: prediction, development of new cytotoxic agents, improvement of the tolerance of normal tissue and the optimisation of radiochemotherapy. MATERIAL AND METHOD: For the development of new strategies the knowledge of molecular mechanisms of radiation induced cell death is essential. In the present article a novel biological strategy for optimisation of radiotherapy is introduced. RESULTS: Distinct pathways mediate apoptosis in response to death receptor stimulation and ionizing radiation although an overlapping set of molecules is involved. TRAIL (TNF alpha related apoptosis inducing ligand) is a type-II membrane protein belonging to the TNF family, which preferentially induces apoptotic cell death in a wide variety of tumor cells but not in normal cells. Based on the assumption of distinct signalling pathways, combination of TRAIL and ionizing radiation seems ideal to increase the therapeutic efficacy. Our study presents the rationale, own data, data of other groups as well as the current status of the latest findings regarding the function and physiological role of the TRAIL death ligand. CONCLUSIONS: Apoptosis resistance towards ionising radiation is not associated with a general apoptosis resistance. Death ligands, for example TRAIL trigger apoptosis even in cells not undergoing apoptosis in response to radiation. A combination of both modalities induces additive or synergistic effects in regard to apoptosis induction and eradication of clonogenic tumor cells. Thus, the combination of TRAIL with radiation proofs that novel treatment strategies may be developed on the basis of molecular or cell biological research. PMID- 12627257 TI - Physical and technical quality assurance in the CHARTWEL-Bronchus trial. AB - BACKGROUND: On-site physical quality assurance (QA) was performed in the participating centers of the CHARTWEL-Bronchus trial to ensure that physical and technical treatment parameters correspond with the requirements of this trial. MATERIAL AND METHODS: Questionnaires were sent to the clinics to obtain information on the equipment and in-house QA policies. In addition, two phantoms with drillings for an ionization chamber were shipped with detailed instructions for CT-based treatment planning of a fixed field (RW3 phantom) and a standardized isocentric 3-field technique (Rando humanoid phantom). Using their routine treatment planning system, the participating centers performed point dose calculations for the isocenters in both phantoms and for defined points in the lungs and the spinal cord of the Rando phantom. During the on-site visit, the doses in these points and the deviation of the actual monitor calibration from the internal reference value of the department were determined. In addition, relevant geometric parameters of the accelerator were checked. RESULTS: In the RW3 phantom, the maximum dose deviations from the prescribed value were 3.5% without correction for the actual monitor calibration and 2.1% after correction. The maximum dose deviation in the isocenter of the Rando phantom was 4.0%. To separate the influence of the treatment planning system on this deviation from other sources, all measurements in the Rando phantom were corrected for the deviations determined in the RW3 phantom. After this correction, the maximum deviation was 3.0% in the isocenter. For the other measurement points, the largest dose deviation of 7% was found in the left lung. Deviations of geometric parameters were negligible in all audited departments. CONCLUSION: The CHARTWEL Bronchus physical QA program revealed a high conformity of geometric and dosimetric parameters and valid dose calculations by the CT-based treatment planning systems in all audited departments. PMID- 12627258 TI - Inhibition of the iNOS pathway in inflammatory macrophages by low-dose X irradiation in vitro. Is there a time dependence? AB - BACKGROUND: Low radiation doses (< or = 1.25 Gy), if applied 6 h before or after stimulation, are known to inhibit the inducible nitric oxide synthase (iNOS) pathway in inflammatory macrophages in vitro. We therefore investigated the time dependence and the underlying molecular mechanism of this effect, since it may be involved in the clinically observed anti-inflammatory and analgesic efficacy of low-dose radiotherapy. MATERIAL AND METHODS: Metabolic activity, nitric oxide (NO) production, iNOS- and hemoxygenase 1-(HO-1-)protein and -mRNA expression by macrophages in vitro after stimulation with LPS/IFN-gamma (0.1 microg ml(-1)/100 U ml(-1)) were investigated. Irradiation was performed at 6, 4, 2 h before and 0, 2, 4, 6 h after stimulation with doses ranging from 0.3 to 10 Gy. For each group, three independent experiments were performed over a period of 30 h with sampling intervals of 3 h. RESULTS: In stimulated macrophages, metabolic activity was not affected by radiation doses up to 10 Gy. A dose-dependent modulation of the cumulative NO production was observed with significant inhibition by low radiation doses < or = 1.25 Gy) and return to control level and even higher concentrations by higher doses (< or = 5 Gy). The degree of inhibition did not show any significant time dependence within the experimental time window used. The iNOS-mRNA expression 3-18 h following stimulation and subsequent irradiation was not affected by doses < or = 1.25 Gy. The iNOS-protein expression 6-24 h following stimulation and subsequent irradiation was reduced by doses < or = 1.25 Gy. By contrast, neither HO-1-protein nor HO-1-mRNA expression at the same time points was influenced by these low doses. CONCLUSION: The inhibitory interference of low radiation doses with the iNOS pathway in inflammatory macrophages appears to be based on radiation effects on the translational and posttranslational control mechanisms of iNOS activity. However, contrary to our working hypothesis this is not related to radiation-induced induction of HO-1 expression and thereby increased degradation of heme which is essential for iNOS activity. Thus, other posttranslational modifications such as the proteasome degradation pathway might be involved. PMID- 12627260 TI - Psychosocial stress in cancer patients during and after radiotherapy. AB - BACKGROUND AND PURPOSE: The aim of this study was to investigate stress in tumor patients by means of a cancer-specific questionnaire in the course of radiotherapy. MATERIAL AND METHODS: Disease-specific aspects of psychosocial stress (Herschbach's Questionnaire on Stress in Cancer Patients, QSC) were self assessed by patients with different tumor types before radiotherapy (ti1), after radiotherapy (ti2), and 6 weeks after the end of radiotherapy (ti3). We investigated 265 of 446 patients (157 male, 108 female; median age 58.6 years) with complete data of ti1-ti3. RESULTS: In the course of investigation, the most prominent stress scale of the patients proved to be physical efficiency, without significant changes during treatment and after therapy. Significant increases in stress were observed for anxiety, pain, and information at ti3 (p < 0.001, p = 0.001, p = 0.035). Women showed significantly higher stress from ti1 to ti3, younger patients displayed a decrease in anxiety, whereas elderly patients demonstrated an increase (p = 0.016). Breast cancer patients had the highest stress levels. The probability of correctly predicting increase in stress (sensitivity) was 78% and the specificity 67%. The relevant predictor variables were tumor stage, addiction to alcohol or nicotine, metabolic disorder, marital status, and age. CONCLUSION: Patients who experienced stress at the beginning of radiotherapy also had the same or increased levels of stress during and shortly after treatment and needed permanent psychosocial support to improve quality of life. The identification of patients with high stress levels at the beginning of therapy could be helpful. PMID- 12627261 TI - [Radiotherapy in benign diseases: Morbus Peyronie]. AB - PURPOSE: The induratio penis plastica (IPP) or the so-called Morbus Peyronie is a burdening disease for patients with three main symptoms, induration, deviation and pain. The etiology of this fibrosing and plaque forming disease is largely unknown up to now. PATIENTS AND METHODS: The presented data refer to a retrospective univariant examination of 67 patients, which have been treated for IPP with soft X-rays in the Dermatological Clinic of the Technical University of Munich between 1990 and 1995. The aim of the study was to examine, how far a progression of the disease can be stopped with soft X-rays and how the pain symptomatic is reduced. RESULTS: In 58 of 67 examined patients (86.6%) a progression of the disease could be stopped. 25 out of 67 patients (37.3%) complained of pain before therapy. This symptomatic mostly improved totally in 21 patients (84% of the patients with pain). A complete or partial dissolution of the indurations was to be noticed in 41 of 70 indurations (58.6%). In 23 of 60 patients (38.3%) an improvement or totally regression of the deviation was observed. It could be shown that therapeutic outcome significantly correlates to a shorter duration of anamnesis (p < 0.05), smaller plaque size (p < 0.025) and a tendency to lower age of the patients. CONCLUSION: Radiotherapy of Morbus Peyronie is extremely effective and has a low rate of side effects. PMID- 12627259 TI - Amifostine as radioprotective agent for the rectal mucosa during irradiation of pelvic tumors. A phase II randomized study using various toxicity scales and rectosigmoidoscopy. AB - AIM: To evaluate the cytoprotective effect of amifostine against radiation induced acute toxicity to the rectal mucosa. PATIENTS AND METHODS: 36 patients irradiated for prostate or gynecologic cancer were randomized to receive amifostine (n = 18, group A) or not (n = 18, group B). The radiation-induced acute rectal toxicity was evaluated by using three different toxicity scales: WHO scale, EORTC/RTOG toxicity criteria, and a modified toxicity scale based on the LENT-SOMA grading scale and the endoscopic terminology of the World Organization for Digestive Endoscopy. The objective measurements were coming from flexible rectosigmoidoscopy performed at baseline and 1-2 days after completion of the radiotherapy schedule. Anterior-posterior fields were used in the gynecologic patients while 3-D conformal 4-field technique was applied in the prostate cancer patients. The area under the curve (AUC) for dose-volume histograms (DVHs) of the rectum was also assessed during a 3-D treatment planning schedule, and no significant differences were assessed between the two groups, indicating a homogeneous dose-volume effect. RESULTS: Amifostine was well tolerated. No grade 2 or higher WHO and EORTC/RTOG acute toxicity was noted in group A, while acute rectal toxicity (> or = grade 1) was observed in 16/18 patients of group B versus 2/18 of group A (p < 0.001). The onset as well as the duration of acute rectal toxicity were significantly improved in group A (p = 0.002). Rectosigmoidoscopy revealed more severe rectal mucositis in noncytoprotected patients (group B), and modified LENT-SOMA overall mucositis grading score was significantly lower in group A (p = 0.003). CONCLUSION: Amifostine seems to have a significant cytoprotective efficacy in acute radiation-induced rectal mucositis in terms of symptomatic and objective endpoints. PMID- 12627262 TI - Irradiation for conjunctival granulocytic sarcoma. AB - CASE HISTORY AND FINDINGS: A 73-year-old woman with a history of myeloproliferative syndrome (MPS) presented with bilateral chemosis, redness and burning of the eyes. The ocular motility was severely impaired. Ophthalmological examination revealed markedly distended conjunctivas on both sides. Biopsy disclosed conjunctival granulocytic sarcoma as an initial symptom of acute myelogenous leukemia (AML). Diagnosis was confirmed by peripheral blood smear and bone marrow aspiration. TREATMENT AND OUTCOME: The orbital tumor disappeared completely after local external beam irradiation with a total dose of 30 Gy and no further orbital recurrence occurred. With chemotherapy following irradiation transient hematological remission was achieved. 5 months after diagnosis the patient died of respiratory failure following atypical pneumonia as a consequence of her underlying disorder. CONCLUSION: Detection of orbital granulocytic sarcoma, even in the absence of typical leukemic symptoms is of practical importance, because treatment with irradiation can lead to stabilization or improvement in the patient's vision. PMID- 12627263 TI - Comparison between in vivo dosimetry and barium contrast technique for prediction of rectal complications in high-dose-rate intracavitary radiotherapy in cervix cancer patients. AB - PURPOSE: To investigate the correlation between late rectal complications and rectal dose in cervix cancer patients treated with high-dose-rate intracavitary radiotherapy (HDR ICR) and to analyze factors reducing rectal complications. PATIENTS AND METHODS: A total of 136 patients with cervix cancer who were treated with external beam radiotherapy (EBRT) and HDR ICR from 1995 to 1999 were retrospectively analyzed. Radiotherapy (RT) consisted of EBRT plus HDR ICR. The median EBRT dose was 50.4 Gy, and midline block was done after 30-50 Gy of EBRT. A total of six fractions of HDR ICR with 4 Gy fraction size each were applied twice per week to the A point. The rectal dose was calculated at the rectal reference point using the barium contrast criteria. In vivo measurement of the rectal dose was performed with thermoluminescent dosimeter (TLD) during HDR ICR. The median follow-up period was 26 months (range 6-60 months). RESULTS: A total of 16 patients (12%) experienced rectal bleeding, which occurred 4-33 months (median 11 months) after the completion of RT. The calculated rectal doses did not differ in patients with rectal bleeding and those without, but the measured rectal doses were higher in affected patients. The differences of the measured ICR fractional rectal dose, ICR total rectal dose, and total rectal biologically equivalent dose (BED) were statistically significant. When the measured ICR total rectal dose exceeded 16 Gy, the ratio of the measured rectal dose to A point dose was > 70%; when the measured rectal BED exceeded 110 Gy(3), a high possibility of late rectal complications could be found. CONCLUSION: In vivo dosimetry using TLD during HDR ICR was a good predictor of late rectal complications. Hence, if data from in vivo dosimetry shows any possibility of rectal bleeding, efforts should be made to reduce the rectal dose. PMID- 12627264 TI - Locoregional failure 15 years after mastectomy in women with one to three positive axillary nodes with or without irradiation the significance of tumor size. AB - BACKGROUND: There is insufficient evidence to suggest the routine use of postmastectomy radiotherapy (PMRT) in patients with one to three positive axillary nodes and T1/2 tumors. We have assessed the risk of locoregional recurrence (LRR) with or without RT in this group of patients, and focused on the results in subgroups defined by tumor size. PATIENTS AND METHODS: 249 women with T1/2 tumors and one to three positive nodes underwent mastectomy and axillary dissection between 1983 and 1987. Locoregional RT of 50 Gy was given to 175 patients. Chemotherapy or hormonal therapy was administered to 41 and 71 women, respectively. The median follow-up time of survivors was 189 months (range, 167 227 months). RESULTS: The rates of isolated LRR without or with RT were 16% (12/74) and 8% (14/175), respectively (p = 0.05), and the total (with or without distant relapse) LRR rates 23% and 12%, respectively (p = 0.03). 15-year overall survival amounted to 41% without RT and to 52% with RT (p = 0.2). The rates of isolated LRR for patients treated with chemotherapy or hormonal therapy only were 25% and 12%, respectively. In the absence of RT, age (> 45 vs = 45 years; p = 0.06), tumor size (T1 vs T2; p = 0.07), and extranodal invasion (ENI; absent vs present; p = 0.09) were related to the risk of developing an isolated LRR. On multivariate analysis, only tumor size (relative risk [RR], 3.92; 95% confidence interval [CI], 1.11-15.14) and age (RR, 3.37; 95% CI, 1.03-11.09) emerged as independent significant predictors, whereas ENI (RR, 1.50; 95% CI, 0.81-2.77) did not. In the T1 subgroup, the estimated 15-year isolated LRR rate was 9% (3/36) without and 9% (8/99) with RT (p = 0.9775). 15-year disease-free survival amounted to 62% and 57%, respectively (p = 0.5153). For patients without RT, according to the age groups (= 45 vs > 45 years), the 15-year rates of isolated LRR were 9% and 9%, respectively (p = 0.9910). In the T2 subgroup, the estimated 15-year isolated LRR rate was 30% (9/38) without and 10% (6/76) with RT (RR, 0.33; 95% CI, 0.12-0.92; p = 0.0244). 15-year disease-free survival amounted to 32% and 50%, respectively (p = 0.1213). For patients without RT, according to the age groups (< or = 45 vs > 45 years), the 15-year rates of isolated LRR were 57% and 16%, respectively (p = 0.0049). CONCLUSION: Patients with T1 tumor and one to three positive nodes are at low risk of isolated LRR either with or without RT. Patients with T2 tumor and one to three positive nodes are at high risk of isolated LRR without RT. Our findings support the routine use of PMRT in patients with T2 tumor, especially those aged < or = 45 years. PMID- 12627266 TI - Transmyocardial laser revascularisation and other treatment modalities for angina pectoris. AB - Ischaemic heart disease is one of the leading causes of morbidity and mortality in the western world. This paper provides an overview of the different treatments for one of the most common manifestations of ischaemic heart disease: angina pectoris. Besides the currently available conventional methods, several alternative treatments are described, with a special focus on transmyocardial laser revascularisation. PMID- 12627267 TI - The old spontaneously hypertensive rat as a model for transmyocardial laser revascularisation research. AB - Animal models used in transmyocardial laser revascularisation (TMLR) research lack the increased fibrosis observed in patients with chronic myocardial ischaemia. This pathology has also been described in patients with chronic elevated afterload, and therefore we evaluated the spontaneously hypertensive rat (SHR) as a model for TMLR research. We compared (1) the myocardial pathology of SHR with the pathology of three TMLR patients, (2) the reaction to TMLR in SHR and human myocardium using three different lasers, and (3) myocardial hypoxia in SHR myocardium and (healthy) Wistar rat myocardium. SHR and human myocardium both showed increased fibrosis and a similar myocardial reaction to TMLR (comparable morphology of fibrotic TMLR channel scars). More hypoxic cells were observed in SHR than in Wistar control rat myocardium. The similarities between SHR and human chronic ischaemic myocardium make the SHR a suitable model for TMLR research. PMID- 12627268 TI - Coagulation and temperature distribution in Nd: YAG interstitial laser thermotherapy: an in vitro animal study. AB - Interstitial laser thermotherapy (ILTT) was performed increasingly for local destruction of different tumours. The proposal of the present study was the optimisation of the therapy-relevant ILTT parameters and laser application forms which are a prerequisite for the development of an optimal tumour treatment strategy. Laser-induced temperature changes and coagulation patterns with different laser light applicators (bare fibre, ring mode fibre, side fibre, diffuser fibre) were investigated on liver, spleen and tongue tissues of 26 adult pigs after interstitial thermotherapy with Nd: YAG laser. Analysis of experimental results on ILTT and tissue necrosis showed a dependence on laser exposure time, type of laser fibre and applied laser energy. The most homogeneous and extensive coagulation zone was seen after laser treatment with rather low energy over a longer time period. PMID- 12627269 TI - A study on surface roughness and microleakage test in cavities prepared by Er:YAG laser irradiation and etched bur cavities. AB - The purposes of this study were to investigate the surface morphology, suface roughness of cavities prepared by Er:YAG laser irradiation, and compared the microleakage degree after composite resin restoration with etched bur cavities, in vitro. In each of the 30 human extracted teeth, two shallow cavities were prepared; one prepared with the Er:YAG laser system on the buccal surface, and one produced on the lingual (palatal) surface with a high-speed turbine. Five cavities from each group were investigated by scanning electron microscopy (SEM), and five were subjected to surface roughness analysis by a colour laser three dimensional (3D) microscope. The remaining cavities were filled with a composite resin and subjected to a microleakage test under thermocycling. Only bur cavities were acid-etched before filling. Statistical analysis was performed using the Mann-Whitney U test; a value of p <0.01 was considered significant. Morphologically, the prepared cavities showed an irregular surface with the absence of a debris-like smear layer; enamel prisms and opening of dentinal tubules were recognised. Surface roughness was significantly increased with the laser system. Microleakage test revealed no significant differences between the laser and bur cavities. Crosscut sections of the cavities with no microleakage showed no gap at the interface. Laser cavity may facilitate good adaptation of composite resin with enamel and dentine, because an increase of surface roughness and the openings of dentinal tubules may facilitate the formation of a hybrid zone, since a primer and an adhesive can penetrate the surface better when the smear layer is removed. It can be concluded that shallow cavity prepared by Er:YAG laser is capable of decreasing microleakage of composite resin restorations, and its efficiency is similar to etched bur cavities. PMID- 12627270 TI - The possibility of detecting subgingival calculus by laser-fluorescence in vitro. AB - The aim of the present study was to investigate laser-fluorescence effects on the root surface. The possibility of detecting subgingival calculus should be evaluated to control complete calculus removal after scaling and root planing. Twenty freshly extracted human teeth, partially covered with calculus on the root surface, were fixed on a translation stage. The specimens were irradiated by light from a laser source (diode laser, <1 mW) with a wavelength of 655 nm using the Diagnodent trade mark -system. To simulate the conditions in a periodontal pocket during the measurement procedure, the teeth were covered by a layer of physiological saline solution or blood in order to determine the influence of different inflammatory fluids. The root surface was scanned on a line of about 1 cm in steps of 0.5 mm. The start and end was marked by a notch. The results of the laser-fluorescence detection were compared to clinical and histological findings. Clinically apparent calculus on the root surface was always accompanied with a statistically significant increase of laser-fluorescence values ( p < 0.05). These values were not influenced by the different fluids ( p < 0.05). The laser-fluorescence values were reproducible ( p > 0.05). After scaling, when no debris remained at the root surface, laser-fluorescence values decreased significantly ( p < 0.05). The histological findings supported the clinical results. In conclusion, the present study indicates that laser-fluorescence values on the root surface are strongly correlated to the presence of calculus. This fluorescence effect can basically be used to develop a new detecting system for subgingival calculus in periodontal pockets. PMID- 12627271 TI - Wound healing after laser treatment of oral and oropharyngeal cancer. AB - The wound healing after laser surgery is delayed in contrast to conventional scalpel surgery. There are some animal studies, which investigated the effects of laser surgery upon wound healing, whereas clinical investigations on wound healing of human mucous membrane after laser surgery is missing. A histological study on the laryngeal mucosa of 11 patients after laser incisions who underwent emergent laser debulking surgery for acute airway obstruction and a clinical study of 24 patients with oral or oropharynx cancer who were treated by laser surgery were performed to investigate the course of wound healing. The histological findings showed that the beginning of wound healing was delayed after laser surgery. The duration of wound healing after laser surgical tumour resection revealed a clear dependence on the size of the initial defect. The average duration of wound healing after CO(2) laser surgery (32.8 +/- 9.2 days) was significantly shorter than after Nd:YAG laser surgery (40.4 +/- 9.2). Due to the more pronounced zone of necrosis at the base of the wound ground this effect is more evident using the Nd:YAG laser. However, the different course of wound healing with both laser systems does not seem to have a negative influence on functional results. PMID- 12627272 TI - Diode laser modifications for treatment of choroidal neovascularisation. AB - The aim of this paper is to describe various diode laser modifications and their use in treating choroidal neovascularisation in age-related macular degeneration. Diode lasers are used to treat selected choroidal neovascular membranes. Alterations in microprocessor connectivity, and parameters such as maximum spot size, light delivery time and coupled Joule meter, were made so that ophthalmic surgeons could specify treatment possibilities. A trimodal (photocoagulation, transpupillary thermotherapy and photodynamic therapy) application laser device coupled to a single light source has been developed. The new diode laser modifications were technically successful. Microprocessor connectivity was obtained, larger spot sizes were achieved, light delivery time could be extended and energy parameters were available at the display. PMID- 12627273 TI - The laser diffractoscope - a new and fast system to analyse red blood cell flexibility with high accuracy. AB - Red blood cell (RBC) flexibility is one of the determining parameters of the microcirculation. As capillaries are smaller in diameter than RBC, RBC can only pass through by deformation. Reduced flexibility is associated with a reduced microcirculation. In obstetrics, pre-eclampsia and intrauterine growth retardation are regarded as diseases exhibiting a reduced microcirculation. With this new system it might be possible to achieve new data concerning the microcirculation in these diseases. The laser diffractoscope is based on laser light diffraction and computer-assisted image analysis. It offers the ability to determine the flexibility of RBC in whole blood with high accuracy between 0 and 30 Pa. Two hundred microlitres of whole blood are required, and the analysis is carried out in less than 5 min, with a standard deviation < or =1%. The laser diffractoscope is a new system which offers the ability to measure RBC deformability with high accuracy in a short time. Evaluation of reproducibility and validity, as well as comparison with data from other measuring systems, proves the excellence of the laser diffractoscope. An example of its application is shown by analysing the deformability of RBC in 10 uncomplicated pregnancies at term. PMID- 12627274 TI - Bactericidal effects of different laser wavelengths on periodontopathic germs in photodynamic therapy. AB - This study was an attempt to clarify whether the bactericidal effects of photodynamic therapy (PDT) are wavelength or dose-dependent. We also attempted to create an optimised protocol for a light-based bactericidal modality to eliminate periodontal pathogens. Cultures of Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Porphromonas gingivalis, Prevotella intermedia, and Streptococcus sanguis, were exposed to a He-Ne laser (632.8 nm) with a 30 mW power output, a 100 mW diode laser at 665 nm, or a 100 mW diode laser at 830 nm, in the presence or absence of methylene blue (MB) as a photosensitiser. A control group was also used with exposure to MB alone without laser exposure. The cultures were analysed by viable counts. The results indicated that exposure to the 100 mW laser light could eliminate up to 40% of bacteria on average. In particular, the most effective killing occurred with exposure to laser light in combination with the MB photosensitiser. The results of kinetic studies indicated that the best PDT response rate was achieved with a 60 s (energy density 21.2 J/cm(2)) exposure to the 665 nm wavelength diode laser in the presence photosensitiser. In this condition, approximately 95% of A. actinomycetemcomitans and F. nucleatum, and 99-100% of the black-pigmented bacteria ( P. gingivalis and P. intermedia) and S. sanguis were eliminated. These results showed that both wavelength and energy density are important factors, and that a low power laser of optimal wavelength and dosage, in combination with an appropriate photosensitiser, is a practical bactericidal modality. We concluded that using a diode laser of proper power and wavelength to deliver 60 s of irradiation could be a useful adjunct with mechanical debridement in the prevention of the re colonisation of subgingival lesions by pathogenic microorganisms. PMID- 12627275 TI - A spectroscopic study of the photobleaching of protoporphyrin IX in solution. AB - Photodynamic therapy (PDT) has developed into an important new clinical treatment for cancer during the past 30 years. The method is non-invasive and based on the photochemical activity of a photosensitising agent present in cells and tissues. In so-called ALA-PDT, protoporphyrin IX (Pp IX) is induced from aminolaevulinic acid (ALA) applied topically or systemically. It has been shown that Pp IX is photodegraded by a photo-oxidation process and that its photoproducts have a characteristic absorption band around 670 nm, as observed both in solution and in cells incubated with ALA. In this study, the involvement of oxygen in the photobleaching process was verified by studying the effect of oxygen depletion using the freeze-pump-thaw (FPT) method. A solution of Pp IX in dimethylformamide (DMF) was exposed to light in the wavelength region 600-700 nm (peak centred at 620 (+/-25) nm) both in the presence and in the absence of oxygen. The bleaching process was observed by absorbance and fluorescence measurements. Photobleaching was observed in the presence of oxygen, as verified by the build-up of a photoproduct absorbing at 670 nm. When the sample was deoxygenated with the FPT method, the photoproduct absorption peak at 670 nm was missing. These results confirm that the formation of photoprotopor-phyrin is a photo-oxidation process and that no photobleaching takes place in the absence of oxygen. When comparing our results to the studies carried out by N(2) bubbling, the N(2) bubbling seems to be insufficient to remove the oxygen completely from the solution. PMID- 12627276 TI - The potential hazard of laser pointers. AB - We have examined 30 laser pointers that have been removed from the possession of accused persons by police in Scotland. An analysis has shown that 28 were Class 3B (European), with radiated power up to 9.3 mW. We have reviewed the relevance of these findings and this shows that they are potentially hazardous if viewed directly. Transient exposure is unlikely to cause long-term damage but prolonged exposure may cause permanent retinal injury. They may also cause dazzle, the consequences of which could be serious. PMID- 12627279 TI - Prevalence and clinical significance of fever in acute exacerbations of chronic obstructive pulmonary disease. AB - Fever is an important, although not always present, sign in acute exacerbation of chronic obstructive pulmonary disease (AECOPD). The objective of this study was to determine the prevalence of fever and its clinical significance in these episodes. Two hundred seventeen AECOPD hospitalizations were included in a prospective study and were categorized as febrile (FH) or afebrile (AFH), using as a cutoff a rectal temperature measurement of 37.8 degrees C during hospitalization. Eighty-six hospitalizations (39.6%) were identified as AFH. The patients in this group had significantly more severe airway obstruction and hypoxemia, a higher rate of chronic obstructive pulmonary disease (COPD) complications, a higher rate of type 3 exacerbation, a shorter length of hospitalization and a higher rate of readmissions than patients in the FH group. The results of this study show that more than one-third of AECOPD hospitalizations are attributable to afebrile episodes. These episodes are characterized by lower rates of bronchitis manifestations in patients with more severe disease. Although the length of hospitalization for these episodes is shorter, the readmission rate is higher. PMID- 12627280 TI - Respiratory disease due to parainfluenza virus in adult leukemia patients. AB - Reports of human parainfluenza viruses (HPIV) in patients with leukemia have been limited to a few cases or as a portion of general surveys. In order to expand the knowledge of these infections in this patient group, the frequency and clinical course of HPIV infections was determined among 1,787 patients with leukemia treated at The University of Texas M.D. Anderson Cancer Center between July 1994 and December 1997. HPIV was isolated from 47 (6.2%) of the 770 patients who were cultured for respiratory viruses. HPIV type 3 accounted for 39 of the 47 HPIV infections. Twenty-six patients developed pneumonia, and the associated mortality was 27%. Multivariate analysis revealed that a low absolute lymphocyte count and pneumonia were associated with increased mortality. Concurrent respiratory and other infections were associated with an increased frequency of pneumonia. Only five patients with pneumonia received antiviral therapy and four of them survived the infection. HPIV infection in leukemic patients is frequently associated with pneumonia and the mortality rate from pneumonia is substantial among lymphopenic patients. PMID- 12627281 TI - Helicobacter pylori virulence genotypes in Portuguese children and adults with gastroduodenal pathology. AB - The aim of this study was to evaluate the prevalence of virulence genotypes, namely cagA, vacA and babA2, of Helicobacter pylori strains isolated from Portuguese adults and children presenting gastroduodenal pathology. One hundred thirty-six strains were studied, 82 isolated from adult patients (50 with nonulcerative gastritis and 32 with active peptic ulcer) and 58 isolated from children (54 with nonulcerative gastritis and 4 with duodenal ulcer). Genotyping of cagA, vacA and babA2 was assessed by polymerase chain reaction. Overall, Helicobacter pylori strains carrying more virulent genotypes were much more prevalent in adults than in children, particularly the type I ( vacAs1- and cagA positive) and the triple-positive ( vacAs1-, cagA- and babA2-positive) strains ( P<0.001). A subpopulation of adults and children with nonulcerative gastritis was also studied, and differences in the prevalence of virulent genotypes were observed, either for individual genotypes ( P=0.017 for cagA, P=0.010 for vacAs1) or in combinations, i.e. the type I genotype ( P=0.005) and the triple-positive strains ( P=0.031). There was no difference between the two populations in the distribution of babA2 and m1/m2 genotypes. Considering the cohort effect in the epidemiology of Helicobacter pylori infection, these results suggest that different strains might circulate during different periods of time, or that, after infection in childhood, individual strains will undergo changes during the course of infection. PMID- 12627282 TI - Frequent occult hepatitis B virus infection in patients infected with human immunodeficiency virus type 1. AB - The presence of hepatitis B virus (HBV) serological markers was investigated in 170 patients (137 male, 33 female) infected with human immunodeficiency virus (HIV) type 1. Antibodies to the hepatitis B core antigen (anti-HBc antibodies) were detected in 115 (68%) patients. Of these 115, 14 (12%) were hepatitis B surface antigen (HBsAg) positive, 60 (52%) presented anti-HBs antibodies, and 41 (35%) were anti-HBc positive only. All 115 of the anti-HBc positive samples were tested for HBV DNA by using two polymerase chain reaction (PCR) assays that amplify the core and pre-S regions of the HBV genome, respectively. HBV DNA was detected in 23 samples: 7 of 14 (50%) HBsAg-positive samples, 12 of 60 (20%) anti HBs-positive samples, and 4 of 41 (10%) samples positive for anti-HBc only. Six samples (all HBsAg positive) were positive in both PCR assays and 17 samples were HBV DNA positive in only one assay. The mean viral load in HBsAg-positive patients was higher than that observed in HBsAg-negative patients. A number of patients were receiving treatment with lamivudine, a drug that interferes with both HBV and HIV replication. However, neither the rate of HBV DNA positivity nor HBV load was significantly different between patients treated with lamivudine and those not treated with this drug. PMID- 12627283 TI - Viral and bacterial DNA in carotid atherosclerotic lesions. AB - Atherosclerosis is a major health problem in industrialised countries. Several studies have suggested an association exists between certain microorganisms and the development of atherosclerosis. The aim of the study presented here was to assess the presence of viral or bacterial DNA in carotid atherosclerotic lesions. Nucleic acids were extracted from 18 carotid atherosclerotic lesions that had been collected surgically. Polymerase chain reaction was used to screen for specific genomic DNA from Chlamydia pneumoniae, cytomegalovirus and herpes simplex virus types 1 and 2. An original approach, based on the amplification by PCR of conserved bacterial 16S rDNA nucleotide sequences was also used to detect any bacterial species. The amplification product was identified by sequencing. Chlamydia pneumoniae, cytomegalovirus and herpes simplex 2 DNA were not detected in any of the samples. Herpes simplex 1 DNA was detected in 3 of the 18 samples. Genes encoding bacterial 16S rRNA were amplified and sequenced in eight atherosclerotic lesions. DNA sequences were identified by comparison with sequences registered in the GenBank database. These eight carotid atherosclerotic lesions were shown to contain several bacterial species belonging to human flora or the environment. The exact role of these microorganisms in the genesis or development of the atherosclerotic lesions remains unclear, but they may increase the inflammatory process or be an epiphenomenon. PMID- 12627285 TI - Q fever in adults: review of 66 clinical cases. AB - Sixty-six cases of Q fever in adults, serologically confirmed by indirect immunofluorescence, were studied to analyze the epidemiological, clinical and therapeutic aspects of the disease. Eighty-three percent of the patients were male, and the mean age was 44.7 years. Contact with animals was recorded in 24 patients. The main clinical form of presentation was pneumonia (37 cases); eight patients had hypoxia, and five had respiratory failure. The empirical treatment consisted of macrolides in 36% of cases. Evolution was favorable in all cases. PMID- 12627286 TI - Successful control of disseminated Scedosporium prolificans infection with a combination of voriconazole and terbinafine. AB - Disseminated Scedosporium prolificans infections are almost uniformly fatal because of their resistance to antifungal agents. Recently, synergy between triazoles and terbinafine has been demonstrated against Scedosporium prolificans in vitro. Reported here is a patient who developed disseminated Scedosporium prolificans infection following bone marrow transplantation and who was successfully treated with a combination of voriconazole and terbinafine in addition to aggressive surgical debridement. Antifungal synergy testing and combination therapy should be considered in cases of disseminated infection with Scedosporium prolificans. PMID- 12627284 TI - Tricuspid valve endocarditis due to Neisseria cinerea. AB - Reported here is a case of infective endocarditis caused by the saprophytic species Neisseria cinerea. To the best of our knowledge, this etiology has not been documented in the medical literature previously. The patient was an intravenous drug addict who developed tricuspid endocarditis with lung embolism. The disease was cured after treatment with ampicillin/clavulanate that was changed to ceftriaxone after an embolic event. PMID- 12627287 TI - Correlation between cidofovir-associated uveitis and failing immunorestoration during HAART. AB - In order to elucidate the factors related to the development of cidofovir associated uveitis in AIDS patients receiving highly active antiretroviral therapy, the characteristics of 17 AIDS patients with Cytomegalovirus retinitis ( n=10) or progressive multifocal leukoencephalopathy ( n=7) treated with intravenous cidofovir were evaluated. Six cases of cidofovir-associated uveitis occurred among the patients with retinitis. No cases were detected among the patients with progressive multifocal leukoencephalopathy. Stepwise linear regression analysis revealed a significantly higher HIV-viremia level ( P=0.01) and a significantly lower CD4+ cell count ( P=0.009) among cases at the time of uveitis onset in comparison with patients who did not develop uveitis. Thus, cidofovir-associated uveitis seems to occur more frequently in AIDS patients with retinitis in whom highly active antiretroviral therapy fails to restore immunity. PMID- 12627288 TI - Improved sensitivity of the Chlamydia trachomatis Cobas Amplicor assay using an optimized procedure for preparation of specimens. AB - The aim of this study was to evaluate the performance of an improved sample preparation procedure that enhances the sensitivity of the Chlamydia trachomatis Cobas Amplicor assay (Roche, Switzerland). This procedure was developed after it was observed that, in some cases, endocervical swabs positive for Chlamydia trachomatis in a direct immunofluorescence assay were negative in the Chlamydia trachomatis Cobas Amplicor. For this procedure, the initial sample volume was increased over that recommended by the manufacturer, the sample was concentrated by centrifugation, and the yield of the extraction was increased by an additional enzymatic lysis. Five hundred sixteen endocervical swabs from women visiting a family planning clinic were tested in parallel by the Chlamydia trachomatis Cobas Amplicor using the extraction procedure recommended by the manufacturer and the modified in-house sample preparation procedure. Eight samples were positive with both procedures. Seven additional samples were positive with the in-house method only. The results show that the in-house sample preparation procedure markedly improves the test sensitivity of the Chlamydia trachomatis Cobas Amplicor assay. Consequently, the use of this improved screening protocol will facilitate the detection of even low-level infections with Chlamydia trachomatis, which in turn will lead to the introduction of earlier therapeutic interventions. PMID- 12627289 TI - Usefulness of Western blot in serological follow-up of newborns suspected of congenital toxoplasmosis. AB - The goal of the study reported here was to compare the results of Western blot with other serological methods for testing newborns suspected of having congenital toxoplasmosis. Western blot, enzyme-linked immunosorbent assay, immunoglobulin (Ig)M immunosorbent agglutination assay, and indirect immunofluorescence assay were performed on the sera of 126 neonates collected at birth and at 1 and 3 months of life. Western blot was more sensitive than IgM detection with the immunosorbent agglutination assay (82.6% vs. 69.6%), and the specificity of the two methods was 96.1% and 92.2%, respectively. Among the serological techniques tested, the combination of Western blot (IgG and IgM) with IgM immunosorbent agglutination assay achieved the greatest improvement in the sensitivity of early (postpartum) diagnosis of congenital toxoplasmosis. PMID- 12627290 TI - Clinical and laboratory characteristics of 144 patients with mediterranean spotted fever. PMID- 12627291 TI - Evaluation of three commercial assays for the detection of hepatitis a virus. PMID- 12627292 TI - Therapeutic efficacy of linezolid in extended clinical practice. PMID- 12627293 TI - Estimating attributable mortality of candidemia: clinical judgement vs matched cohort studies. PMID- 12627295 TI - Finnish Disease Heritage I: characteristics, causes, background. AB - This review of the Finnish Disease Heritage (FDH), a group of rare hereditary diseases that are overrepresented in Finland, includes the following topics: FDH characteristics, causes and background, primary theory, revis(it)ed theory, consanguineous marriages in Finland, internal migration of the 1500s, family series for further FDH studies, geography and population structure as a basis for FDH, geography of individual diseases, the structure of FDH families, family structure in individual diseases, Finnish gene mutations, linkage disequilibrium and haplotypes, age of gene mutations, frequencies of disease genes and carriers, and a short description of the possible future of FDH. PMID- 12627296 TI - Finnish Disease Heritage II: population prehistory and genetic roots of Finns. AB - In the second part of my review of the Finnish Disease Heritage (FDH), I discuss the settling of Finland; factors influencing the genes of a population, such as agriculture versus hunting/fishing/gathering, trading and cultural relations, wars and other kinds of violence, and bottlenecks; relatives of the Finns in the light of classical European studies, classical Finnish studies, mtDNA and Y chromosomal studies; the genes of the Finns today, characterizing FDH, the east west difference among Finns, and minorities in Finland, viz. the Lapps or Saami and Swedish-speaking Finns. PMID- 12627298 TI - The effect of plyometric training on distance running performance. AB - Previous research has reported that plyometric training improves running economy (RE) and ultimately distance-running performance, although the exact mechanism by which this occurs remains unclear. This study examined whether changes in running performance resulting from plyometric training were related to alterations in lower leg musculotendinous stiffness (MTS). Seventeen male runners were pre- and post-tested for lower leg MTS, maximum isometric force, rate of force development, 5-bound distance test (5BT), counter movement jump (CMJ) height, RE, VO(2max), lactate threshold (Th(la)), and 3-km time. Subjects were randomly split into an experimental (E) group which completed 6 weeks of plyometric training in conjunction with their normal running training, and a control (C) group which trained as normal. Following the training period, the E group significantly improved 3-km performance (2.7%) and RE at each of the tested velocities, while no changes in VO(2max) or Th(la) were recorded. CMJ height, 5BT, and MTS also increased significantly. No significant changes were observed in any measures for the C group. The results clearly demonstrated that a 6-week plyometric programme led to improvements in 3-km running performance. It is postulated that the increase in MTS resulted in improved RE. We speculate that the improved RE led to changes in 3-km running performance, as there were no corresponding alterations in VO(2max) or Th(la). PMID- 12627299 TI - Effects of inducing physiological hyperglucagonemia on metabolic responses to exercise. AB - The purpose of the present study was to assess the effects of exogenously increasing the circulating levels of glucagon on the metabolic responses to exercise in rats. A total of six groups of rats were infused (iv) either with glucagon (20 or 50 ng x kg(-1) x min(-1)) or saline (0.9% NaCl), either in the resting state or during a bout of running exercise (45 min, 26 m x min(-1), 0% grade). Blood samples were taken at the end of the 45-min experiment. Animals infused with glucagon at 50 ng x kg(-1) x min(-1) showed significantly (P<0.01) higher mean plasma glucagon concentrations than animals infused with saline or glucagon at 20 ng x kg(-1) x min(-1). In addition, exercise resulted in significantly (P<0.05) higher mean plasma glucagon concentrations, compared to rest, in all groups. In spite of these differences in glucagon concentrations, there were no significant (P>0.05) effects of exercise and glucagon infusion on mean hepatic glycogen, plasma glucose, insulin, C-peptide, beta-hydroxybutyrate, or catecholamine concentrations. Although exercise resulted in a significant (P<0.01) increase in plasma glycerol and free fatty acid concentrations and a significant (P<0.05) decrease in glycogen in the soleus muscle, these responses were not affected by the glucagon infusion. These results suggest that the liver is non-responsive to physiological hyperglucagonemia in a short-term (45 min) exercise situation. PMID- 12627300 TI - Changes in blood lipid peroxidation markers and antioxidants after a single sprint anaerobic exercise. AB - It has been well demonstrated that the principal factor responsible for oxidative damage during exercise is the increase in oxygen consumption. However, other theoretical factors (acidosis, catecholamine autoxidation, ischemia-reperfusion syndrome, etc.) that are known to induce, in vitro, oxidative damage may also be operative during short-term supramaximal anaerobic exercise. Therefore, we hypothesized that short-term supramaximal anaerobic exercise (30-s Wingate test) could induce an oxidative stress. Lipid peroxidation markers [serum lipid radical production detected by electron spin resonance (ESR) spectroscopy and plasma malondialdehyde (MDA) levels detected by the thiobarbituric acid reactive substances (TBARS) method], as well as erythrocyte antioxidant enzyme activities [glutathione peroxidase (GPx), superoxide dismutase (SOD)] and erythrocyte glutathione (GSH) levels, were measured at rest, after the Wingate test and during the 40 min of recovery. The recovery of exercise was associated with a significant increase (x2.7) in lipid radical production detected by ESR spectroscopy, as well as with changes in the erythrocyte GSH level (-13.6%) and SOD activity (-11.7%). The paradoxical decrease in plasma TBARS (-23.7%) which was correlated with the peak power developed during the Wingate test ( r=-0.7), strongly suggests that such exercise stimulates the elimination of MDA. In conclusion, this study demonstrates that short-term supramaximal anaerobic exercise induces an oxidative stress and that the plasma TBARS level is not a suitable marker during this type of exercise. PMID- 12627301 TI - Cortical bone resorption during exercise is interleukin-6 genotype-dependent. AB - The objective of this study was to examine the relationship between the interleukin-6 (IL-6) -174 G>C promoter polymorphism and exercise-induced femoral cortical bone resorption. Skeletal response to exercise was assessed in 130 male Caucasian army recruits. Five cross-sectional magnetic resonance images of the right femur were obtained before and after a 10-week period of basic physical training, and changes in cross-sectional cortical area were calculated. Recruits were genotyped for the -174 G>C IL-6 promoter polymorphism. Genotype frequencies (GG 36%, GC 47%, CC 22.17%) were in Hardy-Weinberg equilibrium. The mean percentage change in proximal femoral cross-sectional cortical area was strongly IL-6 genotype-dependent, with GG homozygotes losing 6.8 (3.82)% in cortical area, GC gaining+5.5 (4.88)% and CC gaining+17.3 (9.46)% (P=0.007 for linear trend). These changes persisted throughout the right femur and were significant in the femur as a whole (P=0.03). This study demonstrates an association between a functional polymorphism in the IL-6 gene and femoral cortical remodelling during strenuous physical exercise. Previous studies have suggested an important role for IL-6 in the regulation of bone mass in postmenopausal women, and in the invasion of bone by metastatic tumour deposits. These data extend these observations to the regulation of bone mass in healthy males, supporting a fundamental role for IL-6 in the regulation of bone mass and bone remodelling in humans. PMID- 12627302 TI - Effect of creatine supplementation on cardiac muscle of exercise-stressed rats. AB - The role of creatine supplementation in altering the physiological parameters regulating cardiac muscle's functional capacity through the initiation of cardiac hypertrophy and altered contractile protein expression has not been determined. The purpose of this study was to determine the effect of creatine supplementation, with and without exercise stress, on physiological parameters regulating functional capacity through alterations in rat cardiac mass and contractile-protein expression. Thirty male Sprague-Dawley rats were subjected to 30 min of exercise stress 5 days/week for 3 weeks with 2% of total body mass attached to the tail. Animals were randomly assigned to one of four treatment groups: group 1 (Con) received (1 ml/day) sucrose water by intubation tube (n=8); group 2 (Cr) received (1 ml/day) sucrose/creatine solution (n=6); group 3 (EX) received 1 ml/day sucrose water and the exercise stimulus (n=8), and group 4 (Cr/EX) received (1 ml/day) sucrose/creatine solution and the exercise stimulus (n=8). At the conclusion of the 21-day exercise-training period, the heart was collected and weighed for determination of wet weight, total protein, total RNA, and myosin heavy chain protein expression. RNA concentration decreased significantly (13%) in the EX group, but not in the CR/EX group, indicating an interactive effect of creatine and exercise. Total RNA significantly decreased (15%) in the EX group. Protein concentration significantly increased (9%) in the exercising treatments, while total protein did not change. Cardiac myosin heavy chain expression significantly shifted towards a predominant expression of the beta-isoform in the Cr/EX group [54.53% (3.42) beta]. These results indicate an interaction of creatine supplementation and swimming exercise stress that potentially alters cardiac protein synthesis and demonstrates a possible mechanism through which the combination of creatine supplementation and swimming stress stimuli act to alter the physiological parameters regulating cardiac functional capacity. PMID- 12627303 TI - Delayed reoxygenation after maximal isometric handgrip exercise in high oxidative capacity muscle. AB - We hypothesized that after maximal short-term isometric exercise, when O(2) demand is still high and O(2) supply is not fully activated, higher oxidative capacity muscle may exhibit slower muscle reoxygenation after the exercise than low oxidative capacity muscle. Seven healthy male subjects performed a maximal voluntary isometric handgrip exercise for 10 s. The reoxygenation rate after the exercise (Reoxy-rate) in the finger flexor muscle was determined by near infrared continuous wave spectroscopy (NIRcws) while phosphocreatine (PCr) was measured simultaneously by (31)P magnetic resonance spectroscopy. Muscle oxygen consumption (muscle VO(2)) and muscle oxidative capacity were evaluated using the rate of PCr resynthesis post-exercise. The forearm blood flow (FBF) index at the end of exercise was measured using NIRcws. There was a significant positive correlation between the Reoxy-rate, which ranged between 0.53% s(-1) and 12.47% s(-1), and the time constant for PCr resynthesis, which ranged between 17.8 s and 38.3 s (r(2)=0.939, P<0.001). At the end of the exercise, muscle VO(2) exceeded the resting level by approximately 25-fold, while the FBF index exceeded the resting level by only 3-fold on average. The Reoxy-rate closely correlated with muscle VO(2) (r(2)=0.727, P<0.05), but not with the FBF index. Also, the estimated O(2) balance (muscle VO(2) index/FBF index) was negatively correlated with the Reoxy-rate (r(2)=0.820, P<0.001). These results support our hypothesis that higher oxidative capacity muscle shows slower muscle reoxygenation after maximal short-term isometric exercise because the Reoxy-rate after this type of exercise may be influenced more by muscle VO(2) than by O(2) supply. PMID- 12627304 TI - Neuromuscular adaptations during concurrent strength and endurance training versus strength training. AB - The purpose of this study was to investigate effects of concurrent strength and endurance training (SE) (2 plus 2 days a week) versus strength training only (S) (2 days a week) in men [SE: n=11; 38 (5) years, S: n=16; 37 (5) years] over a training period of 21 weeks. The resistance training program addressed both maximal and explosive strength components. EMG, maximal isometric force, 1 RM strength, and rate of force development (RFD) of the leg extensors, muscle cross sectional area (CSA) of the quadriceps femoris (QF) throughout the lengths of 4/15-12/15 (L(f)) of the femur, muscle fibre proportion and areas of types I, IIa, and IIb of the vastus lateralis (VL), and maximal oxygen uptake (VO(2max)) were evaluated. No changes occurred in strength during the 1-week control period, while after the 21-week training period increases of 21% (p<0.001) and 22% (p<0.001), and of 22% (p<0.001) and 21% (p<0.001) took place in the 1RM load and maximal isometric force in S and SE, respectively. Increases of 26% (p<0.05) and 29% (p<0.001) occurred in the maximum iEMG of the VL in S and SE, respectively. The CSA of the QF increased throughout the length of the QF (from 4/15 to 12/15 L(f)) both in S (p<0.05-0.001) and SE (p<0.01-0.001). The mean fibre areas of types I, IIa and IIb increased after the training both in S (p<0.05 and 0.01) and SE (p<0.05 and p<0.01). S showed an increase in RFD (p<0.01), while no change occurred in SE. The average iEMG of the VL during the first 500 ms of the rapid isometric action increased (p<0.05-0.001) only in S. VO(2max) increased by 18.5% (p<0.001) in SE. The present data do not support the concept of the universal nature of the interference effect in strength development and muscle hypertrophy when strength training is performed concurrently with endurance training, and the training volume is diluted by a longer period of time with a low frequency of training. However, the present results suggest that even the low-frequency concurrent strength and endurance training leads to interference in explosive strength development mediated in part by the limitations of rapid voluntary neural activation of the trained muscles. PMID- 12627305 TI - The use of magnetic resonance imaging to evaluate the effects of cooling on skeletal muscle after strenuous exercise. AB - The purpose of this study was to investigate the separate effects of cooling during the acute (within 60 min post-exercise) or subacute (24-168 h post exercise) phase on skeletal muscle after exercise. Twenty-eight male subjects [mean (SD) 23.8 (1.8) years] were randomly assigned to the control (COTG, n=10), cold-water immersion (CWIG, n=9), and double-cold-water immersion groups (DCWIG, n=9). The cold-water immersion (15 min) was administered to the subjects' legs after calf-raise exercise (CWIG: after recording initial post-exercise measures, DCWIG: after recording initial and 24 h post-exercise measures). Magnetic resonance T2-weighted images were obtained to calculate the T2 relaxation time (T2) of the triceps surae muscle before, immediately after, and at the following times post-exercise: 20, 40, and 60 min, and 24, 48, 96 and 168 h. In addition, the ankle joint range of motion, serum creatine kinase and lactate dehydrogenase, and muscle soreness level were investigated before and after exercise. In all groups, significant T2 elevations in the gastrocnemius muscle appeared from immediately after to 60 min after exercise (P<0.05). Thereafter, COTG showed significantly re-elevated T2 levels in the gastrocnemius at 96-168 h post exercise (P<0.05), while CWIG and DCWIG showed significantly smaller T2 values than the COTG at 96 h post-exercise (P<0.05). In addition, COTG showed larger increases in serum enzymes at 96 h post-exercise (not significant) and significantly greater muscle soreness levels at 48 h post-exercise (P<0.05) than the cooling groups. The results of this study may suggest that cooling has no dramatic effect, but some minor effects on reducing exercise-induced muscle edema in the subacute phase and relieving the extent of the damaged muscle cells. PMID- 12627306 TI - Prolonged exercise potentiates sarcoplasmic reticulum Ca2+ uptake in rat diaphragm. AB - The effects of a single bout of prolonged treadmill exercise [mean=81 (13) min] on sarcoplasmic reticulum (SR) Ca(2+) release, uptake and ATPase activity were determined in the costal region of rat diaphragm (D) and red gastrocnemius (RG). Glycogen depletion measurements made immediately following exercise suggested that treadmill running substantially recruited the fibers throughout both muscles. SR Ca(2+) ATPase activity, measured in isolated SR vesicles, decreased in the RG by 33% but remained unchanged in D in response to the exercise bout. This effect in RG was matched by a 37% decline in Ca(2+) uptake and a 28% depression in Ca(2+) release when measured in muscle homogenates. Conversely, Ca(2+) uptake increased between 157% and 263% in the D in the absence of any change in Ca(2+) release. These data show that the attenuation of SR function that has been consistently observed in limb muscle over the last several decades is absent in diaphragm despite the fact that its fibers appear to experience sufficient activity to deplete their glycogen. In fact, the large increase in Ca(2+) uptake in D shows that prolonged activity actually potentiates the ability of SR vesicles to sequester Ca(2+) in the absence of any increase in energy cost. Thus, it appears necessary to re-evaluate the role of exercise in regulating Ca(2+) sequestration by the SR as different muscles may respond in ways that are dictated by their function. PMID- 12627307 TI - Effects of exercise on vasodilatory capacity in endurance- and resistance-trained men. AB - To determine vasodilatory responsiveness we measured forearm blood flow (FBF) following reactive hyperemia (RH), prior to and following a bout of maximal aerobic exercise in endurance- (n=14) and resistance-trained men (n=10). Both groups were similar in height, body mass, and percentage body fat. Using strain gauge plethysmography, resting FBF was higher in the resistance-trained group [4.82 (0.84) vs 3.33 (1.17) ml min(-1) 100 ml(-1) of tissue; P<0.05]. However, the resistance-trained group had a 17%-29% lower pre-exercise FBF response to RH for the first 45 s (P<0.05). Following the maximal exercise bout there were no group differences in FBF. Post-exercise FBF was higher compared to pre-exercise values in both the endurance- (P<0.001) and resistance- (P<0.01) trained groups. Endurance-trained men appear to have a greater peak vasodilatory capacity compared to resistance-trained men, and acute maximal exercise increased the vasodilatory capacity in both groups. Acute exercise also equalized the peak vasodilatory response between the endurance- and resistance-trained groups, suggesting the potential for flow-mediated vasodilatation was similar for both groups. PMID- 12627308 TI - Maximal dynamic expiratory pressures with fast and slow inspirations. AB - Maximal dynamic expiratory pressures are higher when forced expiration is preceded by a fast inspiration to total lung capacity (TLC) than when preceded by a slow inspiration and a few seconds pause at TLC. We hypothesized that these pressure differences are due to the stretch-shorten cycle (SSC), which refers to enhancement of muscle force when a concentric muscle contraction is immediately preceded by an eccentric contraction. Seven volunteers [36 (2) years; mean (SEM)] performed maximal forced expirations against minimal resistance with fast (F) or slow (S) maneuvers. F maneuvers consisted of a fast inspiration to TLC followed immediately by a fast expiration, whereas S consisted of a slow inspiration to TLC and a 4- to 5-s pause at TLC prior to forced expiration. We measured esophageal pressure ( P(es)), peak expiratory flow rate (PEFR), and the EMG activity of the transversus abdominis (Tr) by means of intramuscular fine-wire electrodes. The subjects performed several runs of each maneuver in a random order, and runs with the greatest expiratory P(es) were analyzed. In comparison with S, F yielded greater P(es) [182 (15) versus 167 (15) cm H(2)O; P=0.003)] but similar PEFR [9.8 (0.7) versus 9.6 (0.7) l/s, P>0.05] and EMG activity of the Tr during forced expiration [221 (31) versus 208 (34) a.u., P>0.05]. Further analysis revealed significant EMG activity of Tr during end-inspiration (eccentric contraction) with F maneuvers only [73 (22) versus 32 (17) a.u., P<0.05]. We conclude that the ability of expiratory muscles to generate greater P(es) with F maneuvers is related to the sequence of an eccentric contraction, which is followed immediately by concentric contraction in a manner analogous to SSC described in skeletal muscles. PMID- 12627309 TI - Surface electromyograms of agonist and antagonist muscles during force development of maximal isometric exercises--effects of instruction. AB - Surface integrated electromyograms (iEMG) of agonist and antagonist muscles were studied during the rising phase of maximal isometric efforts (elbow flexion, unilateral and bilateral leg extension) to explain the difference in maximal rate of force development (MRFD) with a hard-and-fast instruction (instruction I) and a fast instruction (instruction II ). Force and EMG were simultaneously recorded in 24 athletes and iEMG were computed at MRFD and during different phases of force development (P(0-25), P(25-50), P(50-75), P(75-90) and P(90-100)). A two way ANOVA for repeated measures (muscle x instruction) showed that the value of iEMG at MRFD was significantly higher with instruction II for elbow flexion, unilateral and bilateral leg press exercises (F>4.9; P<0.04). The effect of instruction upon iEMG of the agonist muscles corresponding to the different phases of the force development was significant for elbow flexion (F=4.2; P<0.05 ) unilateral (F>6.4; P<0.02) and bilateral leg extension (F>9 and P<0.006 for soleus and vastus lateralis; but F=3.2 and P=0.08 for vastus medialis). There was a significant interaction between instruction and phase of force development (F>2.6; P<0.05 ): iEMG was significantly higher with instruction II at the beginning of force development (P(0-25)) for all the muscles (except the soleus muscle during the bilateral leg exercise) but not at higher force (P(75-90) and P(90-100)). The steeper force development with instruction II can be explained by the better activation of the agonist muscles at the beginning of force development. A lower co-activation of the antagonist muscles does not explain the improvement in MRFD as the iEMG of the antagonist muscles was not lower with instruction II but was proportional to the activation level of the agonist muscle. PMID- 12627310 TI - Effects of dynamic resistance training on heart rate variability in healthy older women. AB - Twenty healthy women aged between 65 and 74 years, trained three times a week, for 16 weeks, on a cycle ergometer, to determine the effects of dynamic resistance training on heart rate variability (HRV). Subjects were allocated to two training groups, high (HI, n=10) and low (LO, n=10) intensity. The HI group performed eight sets of 8 revolutions at 80% of the maximum resistance to complete 2 pedal revolutions (2RM); the LO group performed eight sets of 16 pedal revolutions at 40% of 2RM. Subjects were tested twice before, as control period ( 4 weeks and 0 weeks) and once after training (16 weeks) for HRV, maximum voluntary contraction (MVC) of knee extensors and peak power (P(p)) of lower limbs by jumping on a force platform. HRV was measured using time and frequency domain parameters. Two-way ANOVA for repeated measures was performed on all variables (P<0.05). Results showed no differences between training groups. Following training HRV was not modified, while MVC and P(p) significantly increased. The two proposed forms of dynamic resistance training were appropriate to improve muscle power and strength in elderly females without affecting HRV. More research should verify the effects of an isometric and more prolonged training stimulus on HRV in older subjects. PMID- 12627311 TI - Influence of hypoxia on coordination between breathing and cycling rhythms in women. AB - To investigate interactions between neural (movement) and chemical (hypoxia) respiratory drives during exercise, we analyzed coordination between breathing and cycling rhythms in normoxia (N) and hypoxia (H, 14.5% O(2)). Twenty women [28 (1) years old] cycled for 6 min at three workloads (55%, 75%, and 95% peak oxygen consumption, VO(2peak); WL1, WL2, and WL3) in N and H. Leg movements, respiratory parameters, peripheral oxygen saturation, and heart rate were continuously recorded. The degree of coordination (%COORD) was quantified as the percentage of breaths starting during the same phase of leg movement. Ventilatory response to isocapnic hypoxia (VRH) was assessed at rest during an exposure to an end-tidal PO(2) of 50 mmHg. There were no differences in %COORD between N and H at any of the three workloads, but %COORD increased significantly from WL1 to WL3 in H. There was no correlation between VRH and %COORD. In conclusion, chemical and neural respiratory drives did not competitively interact: coordination between breathing and cycling rhythms was not reduced during H and did not depend on individual VRH. PMID- 12627312 TI - Methodological aspects of maximal lactate steady state-implications for performance testing. AB - The maximal lactate steady state (MLSS) is the highest blood lactate concentration (BLC) that can be identified as maintaining a steady-state during a prolonged submaximal constant workload. Comparative interpretation of published data about MLSS is complicated by the fact that different methods of testing have been utilized. Thus, three methods, corresponding to the time course of changes in BLC incurred during either 30 min (MLSS I) or 20 min (MLSS II and III) of constant submaximal workload exercise, were compared in 26 male subjects [mean (SD) age 24.6 (5.6) years, height 181.6 (4.9) cm, body mass 74.4 (5.2) kg]. MLSS I [5.1 (1.3) mmol x l(-1)], II [4.9 (1.3) mmol x l(-1)], and III [4.3 (1.3) mmol x l(-1)] were different (P<0.01). The workload corresponding to MLSS III [244.8 (44.0) W] was lower (P<0.01) than that at MLSS I [254.0 (40.8) W] and II [251.9 (40.4) W]. No difference could be confirmed between the workloads established for MLSS I and MLSS II. The differences between MLSS I, MLSS II, and MLSS III and corresponding workloads reflect insufficient contribution to lactate kinetics by testing procedures that depend strongly upon the time course of changes in BLC during the initial 20-25 min of constant-workload exercise. Based on the present findings, constant-load tests lasting at least 30 min and a BLC increase of no more than 1.0 mmol x l(-1) after the 10th testing minute appear to be the most reasonable with respect to valid testing results. PMID- 12627313 TI - Influence of vitamin C supplementation on oxidative and salivary IgA changes following an ultramarathon. AB - This randomized study measured the influence of vitamin C ( N=15) compared to placebo ( N=13) supplementation on oxidative and salivary immunoglobulin A (sIgA) changes in runners competing in an ultramarathon race. Seven days prior to the race, subjects ingested in randomized, double-blind fashion three 500-mg tablets of vitamin C or placebo each day. On race day, blood and saliva samples were collected 1 h pre-race, after 32 km of running, and then again immediately post race. During the race, runners received 1 l/h carbohydrate beverages (60 g/l) with vitamin C (150 mg/l) or without in a double-blinded fashion. The runners also ingested two to three carbohydrate gel packs per hour (25 g each). Subjects in both groups ran a mean of 69 km (range 48-80 km) in 9.8 h (range 5-12 h) and maintained an intensity of approximately 75% maximal heart rate (HR(max)) throughout the ultramarathon race. Plasma ascorbic acid was higher in the vitamin C compared to placebo group pre-race, and increased significantly in the vitamin C group during the race [post-race, 3.21 (0.29) and 1.28 (0.12) microg/100 microl, respectively, P<0.001]. No significant group or interaction effects were measured for lipid hydroperoxide and F(2)-isoprostane, but both oxidative measures rose significantly during the ultramarathon race. Saliva volume, sIgA concentration, sIgA secretion and sIgA:saliva protein ratio all decreased significantly (P<0.001) during the race, but the pattern of change in all saliva measures did not differ significantly between groups. No significant correlations were found between post-race plasma vitamin C, oxidative, and saliva measures, except for a positive correlation between post-race serum cortisol and serum vitamin C (r=0.50, P=0.006). These data indicate that vitamin C supplementation in carbohydrate-fed runners does not serve as a countermeasure to oxidative and sIgA changes during or following a competitive ultramarathon race. PMID- 12627297 TI - The Finnish Disease Heritage III: the individual diseases. AB - This article is the third and last in a series entitled The Finnish Disease Heritage I-III. All the 36 rare hereditary diseases belonging to this entity are described for clinical and molecular genetic purposes, based on the Finnish experience gathered over a period of half a century. In addition, five other diseases are mentioned. They may be included in the list of the "Finnish diseases" after adequate complementary studies. PMID- 12627315 TI - Evaluation of the new ADA and WHO criteria for classification of diabetes mellitus in young adult people (15-34 yrs) in the Diabetes Incidence Study in Sweden (DISS). AB - AIMS/HYPOTHESIS: We aimed to evaluate how an aetiology-based classification, as recommended in the ADA and WHO guidelines for classification of diabetes mellitus, matches clinical judgement in the Diabetes Incidence Study in Sweden (DISS), a study covering incident cases of diabetic patients aged 15 to 34 years. METHODS: During a 1-year period (1998), blood samples were taken at diagnosis and 4 months (median) thereafter. Patients were classified according to clinical judgement by the reporting physicians and assessments of islet antibodies (ICA, GADA, and IA-2A) and plasma C-peptide. RESULTS: In 1998, 422 patients were registered in DISS. Among the 313 patients participating in the follow-up, most with clinical Type 1 diabetes (185/218, 85%, 95% CI 79-89%) were islet antibody positive (ab+) at diagnosis. In addition, 14 out of 58 (24%, 14-37%) with clinical Type 2 diabetes and 21 out of 37 (57%, 40-73%) with unclassifiable diabetes were antibody positive at diagnosis. Further to this, 4 out of 33 (12%, 3-28%) were antibody negative with clinical Type 1 diabetes and 4 out of 44 (9%, 3-22%) with Type 2 had converted to antibody positive at follow-up. Among those who were constantly antibody negative, 10 out of 29 (34%, 18-54%) with clinical Type 1 and 1 out of 16 (6%, 0-30%) with unclassifiable diabetes had fasting plasma C-peptide concentrations below the normal range (<0.25 nmol/l) at follow up. CONCLUSION/INTERPRETATION: Most young adults with clinical Type 1 diabetes (199/218, 91%) had objective Type 1 (ab+ at diagnosis/follow-up and/or low fasting plasma C-peptide concentrations at follow-up), as did one third (18/58, 31%) with clinical Type 2 diabetes and more than half (22/37, 59%) with unclassifiable diabetes. About 10% of those who were antibody negative converted to antibody positive. Our study underlines that a classification considering aetiology is superior to clinical judgement. PMID- 12627316 TI - High prevalence of undiagnosed diabetes mellitus in Southern Germany: target populations for efficient screening. The KORA survey 2000. AB - AIMS/HYPOTHESIS: To estimate the prevalence of undiagnosed diabetes mellitus, impaired glucose tolerance (IGT) and impaired fasting glucose (IFG), and their relations with cardiovascular risk factors in the general population aged 55 to 74 years in Southern Germany. METHODS: Oral glucose tolerance tests were carried out in a random sample of 1353 subjects aged 55 to 74 years participating in the KORA (Cooperative Health Research in the Region of Augsburg) Survey 2000. Prevalences of glucose tolerance categories (1999 WHO criteria) were adjusted for sample probabilities. The numbers needed to screen (NNTS) to identify one person with undiagnosed diabetes were estimated from age-adjusted logistic regression models. RESULTS: Sample design-based prevalences of known and unknown diabetes, IGT, and IFG were 9.0%, 9.7%, 16.8%, 9.8% in men, and 7.9%, 6.9%, 16.0%, 4.5% in women, respectively. In both sexes, participants with undiagnosed diabetes had higher BMI, waist circumference, systolic blood pressure, triglycerides, uric acid, and lower HDL-cholesterol than normoglycaemic subjects. A combination of abdominal adiposity, hypertension, and parental diabetes in men resulted in a NNTS of 2.9 (95%CI: 2.0-4.6). In women, the combination of increased triglycerides, hypertension and parental diabetes history yielded a NNTS of 3.2 (95%CI: 2.2-5.1). CONCLUSION/INTERPRETATION: About 40% of the population aged 55 to 74 years in the Augsburg region have disturbed glucose tolerance or diabetes. Half of the total cases with diabetes are undiagnosed. Cardiovascular risk factors worsen among glucose tolerance categories, indicating the need for screening and prevention. Screening for undiagnosed diabetes could be most efficient in individuals with abdominal adiposity (men), hypertriglyceridaemia (women), hypertension, and parental diabetes history. PMID- 12627314 TI - Control of body weight: a physiologic and transgenic perspective. PMID- 12627317 TI - Early adiposity rebound in childhood and risk of Type 2 diabetes in adult life. AB - AIMS/HYPOTHESIS: Type 2 diabetes is associated with a small body size at birth and a high BMI in adult life. The aim of our study was to assess the associations between Type 2 diabetes and birth size, infant growth and age at adiposity rebound. METHODS: We carried out a longitudinal study of 8760 subjects born in Helsinki during 1934 to 1944. On average, they had 18 measurements of height and weight between birth and 12 years of age. In western countries BMI usually decreases after the age of 2 years and rises again at around 6 years--the so called adiposity rebound. We defined age at adiposity rebound by the age of lowest BMI between one and 12 years. We identified people with Type 2 diabetes using a national register. RESULTS: A total of 290 individuals developed Type 2 diabetes in adult life. The cumulative incidence of Type 2 diabetes decreased progressively from 8.6% in persons whose adiposity rebound occurred before the age of 5 years to 1.8% in those in whom it occurred after 7 years ( p<0.001). Early adiposity rebound was preceded by low weight gain between birth and 1 year ( p<0.001). CONCLUSION/INTERPRETATION: Large differences in the incidence of Type 2 diabetes are associated with growth rates in utero, weight gain in infancy and age at adiposity rebound. PMID- 12627318 TI - Loss of beta-cell mass leads to a reduction of pulse mass with normal periodicity, regularity and entrainment of pulsatile insulin secretion in Gottingen minipigs. AB - AIMS/HYPOTHESIS: Type 2 diabetes is associated with impaired insulin action and secretion, including disturbed pulsatile release. Impaired pulsatility has been related to impaired insulin action, thus providing a possible link between release and action of insulin. Furthermore, progressive loss of beta-cell mass has been implicated in the pathogenesis of Type 2 diabetes. The aim of this study was to evaluate a possible link between loss of beta-cell mass and impaired pulsatile insulin secretion with special focus on glucose responsiveness of insulin secretion. METHODS: The kinetic and dynamic profiles of insulin in Gottingen minipigs are favourable for studies on pulsatility and a model of diabetes with reduced beta-cell mass has recently been established. Pigs were studied before (n=14) and after (n=10) reduction of beta-cell mass by nicotinamide (67 mg/kg) and streptozotocin (125 mg/kg) from 17.7+/-4.7 (normal animals, n=5) to 6.1+/-2.0 mg/kg. Pulsatile insulin secretion was examined during basal (n=8 normal, n=6 beta-cell reduced) and glucose entrained (n=6 normal, n=4 beta-cell reduced) conditions. Insulin concentration time series were analysed by autocorrelation and spectral analyses for periodicities and regularity, and by deconvolution for pulse frequency, mass and amplitude. RESULTS: Reduction of beta cell mass and secondary hyperglycaemia resulted in correspondingly (r=0.7421, p=0.0275) reduced pulse mass (42% of normal during basal and 31% during entrained conditions) with normal periodicity (6.6+/-2.2 vs 5.8+/-2.4 min, p=0.50), regularity and entrainability of insulin secretion. CONCLUSION/INTERPRETATION: Neither beta-cell loss, nor 2 weeks of slight hyperglycaemia, as seen in the beta cell-reduced minipig, probably accounts for the disturbed insulin pulsatility observed in human Type 2 diabetes. PMID- 12627320 TI - Intra-axonal recording from large sensory myelinated axons: demonstration of impaired membrane conductances in early experimental diabetes. AB - AIM/HYPOTHESIS: Diabetic neuropathy is accompanied by a range of positive (paresthaesia, dysesthaesia, pain) and negative (hypesthaesia, anesthaesia) neurological symptoms suggesting widespread alterations in axonal excitability. The nature and the mechanisms underlying these alterations in axonal excitability are not well understood. The aim of this study was to examine the extent of changes in membrane properties of an identified neuronal structure-the large myelinated sensory axons in early experimental diabetes in rats. METHODS: Intra axonal microelectrode recordings from large sensory myelinated axons from the isolated sural nerve in short-term streptozotocin-induced diabetic rats were used to study membrane properties using standard current-clamp technique. RESULTS: In addition to decreased conduction velocity we found several differences in physiological properties of sensory axons from diabetic rats: decreased resting membrane potential, decreased single action potential amplitude associated with slower rate of rise and decrease in inward rectification associated with slight alteration in outwardly rectifying conductances indicating impaired potassium conductances. CONCLUSION/INTERPRETATION: These results extend previous indirect evidence that potassium and sodium ionic conductances, most notably the inward rectifier (IR, I(h)), are altered in large sensory axons of diabetic rats. The depression of IR could underly clinical neurological findings in diabetic patients. PMID- 12627319 TI - Glucose oversupply increases Delta9-desaturase expression and its metabolites in rat skeletal muscle. AB - AIM/HYPOTHESIS: Previous studies have shown that prolonged glucose infusion causes insulin resistance and triglyceride accumulation in rat skeletal muscle. In this study, we investigated a possible relationship between insulin resistance and the composition of different accumulated lipid fractions in rat skeletal muscle. METHODS: Continuous glucose infusion was carried out in rats for 7 days. Lipids were extracted from skeletal muscle, separated by thin layer chromatography and fatty acid composition of phospholipids, triglycerides, diglycerides, free fatty acids and cholesterol esters fractions was analysed by gas chromatography. Delta9-Desaturase mRNA was measured by real time polymerase chain reaction. The enzyme activity was measured in the microsomal fractions. RESULTS: Prolonged glucose infusion (5 days) increased the relative content of palmitoleic acid (16:1 N7) several-fold (2.3- to 5.8-fold) in four out of five lipid fractions and enhanced oleic acid (18:1 N9) two-fold in three lipid fractions suggesting increased Delta9-desaturase activity while the content of several polyunsaturated fatty acids was reduced. In parallel, Delta9-Desaturase mRNA contents and enzyme activities in skeletal muscle were increased 10-fold, 75 fold, 2.6-fold and 7.7-fold after 2 and 5 days of glucose infusion, respectively. CONCLUSION/INTERPRETATION: Our results suggest that long-term glucose oversupply induces a rapid increase in Delta9-desaturase expression and enzyme activity in skeletal muscle which leads to fast and specific changes in fatty acid metabolism possibly contributing to the insulin resistance in this animal model. PMID- 12627321 TI - Effects of the novel (Pro3)GIP antagonist and exendin(9-39)amide on GIP- and GLP 1-induced cyclic AMP generation, insulin secretion and postprandial insulin release in obese diabetic (ob/ob) mice: evidence that GIP is the major physiological incretin. AB - AIMS/HYPOTHESIS: This study examined the biological effects of the GIP receptor antagonist, (Pro3)GIP and the GLP-1 receptor antagonist, exendin(9-39)amide. METHODS: Cyclic AMP production was assessed in Chinese hamster lung fibroblasts transfected with human GIP or GLP-1 receptors, respectively. In vitro insulin release studies were assessed in BRIN-BD11 cells while in vivo insulinotropic and glycaemic responses were measured in obese diabetic ( ob/ ob) mice. RESULTS: In GIP receptor-transfected fibroblasts, (Pro(3))GIP or exendin(9-39)amide inhibited GIP-stimulated cyclic AMP production with maximal inhibition of 70.0+/-3.5% and 73.5+/-3.2% at 10(-6) mol/l, respectively. In GLP-1 receptor-transfected fibroblasts, exendin(9-39)amide inhibited GLP-1-stimulated cyclic AMP production with maximal inhibition of 60+/-0.7% at 10(-6) mol/l, whereas (Pro(3))GIP had no effect. (Pro(3))GIP specifically inhibited GIP-stimulated insulin release (86%; p<0.001) from clonal BRIN-BD11 cells, but had no effect on GLP-1-stimulated insulin release. In contrast, exendin(9-39)amide inhibited both GIP and GLP-1 stimulated insulin release (57% and 44%, respectively; p<0.001). Administration of (Pro(3))GIP, exendin(9-39)amide or a combination of both peptides (25 nmol/kg body weight, i.p.) to fasted (ob/ob) mice decreased the plasma insulin responses by 42%, 54% and 49%, respectively (p<0.01 to p<0.001). The hyperinsulinaemia of non-fasted (ob/ob) mice was decreased by 19%, 27% and 18% (p<0.05 to p<0.01) by injection of (Pro3)GIP, exendin(9-39)amide or combined peptides but accompanying changes of plasma glucose were small. CONCLUSIONS/INTERPRETATION: These data show that (Pro(3))GIP is a specific GIP receptor antagonist. Furthermore, feeding studies in one commonly used animal model of obesity and diabetes, (ob/ob) mice, suggest that GIP is the major physiological component of the enteroinsular axis, contributing approximately 80% to incretin-induced insulin release. PMID- 12627322 TI - Insulin down-regulates resistin mRNA through the synthesis of protein(s) that could accelerate the degradation of resistin mRNA in 3T3-L1 adipocytes. AB - AIMS/HYPOTHESIS: Resistin is a peptide secreted by adipocytes and recognized as a hormone that could link obesity to insulin resistance. This study was designed to examine the effect and mechanism(s) of insulin on resistin expression in 3T3-L1 adipocytes. METHODS: Differentiated 3T3-L1 adipocytes were stimulated with insulin and resistin mRNA expression was examined by Northern blot analysis. In some experiments, the insulin signal was blocked by several chemical inhibitors or overexpression of a dominant negative form (Deltap85) of the p85 subunit of phosphatidylinositol 3-kinase (PI 3-kinase). RESULTS: Insulin treatment caused a reduction of resistin mRNA in time-dependent and dose-dependent manners in 3T3-L1 adipocytes. Pre-treatment with PD98059, an inhibitor of extracellular signal regulated kinase 1/2 (ERK1/2) pathway, or SB203580, an inhibitor of p38 mitogen activated protein-kinase (p38 MAP-kinase) pathway, did not influence insulin induced reduction of resistin mRNA. Inhibition of PI 3-kinase by LY294002 or Deltap85 also failed to block insulin-induced reduction of resistin mRNA. Cycloheximide, a protein synthesis inhibitor, completely blocked insulin-induced reduction of resistin mRNA. Actinomycin D, a RNA synthesis inhibitor, also blocked insulin-induced reduction of resistin mRNA, and the decreasing rate of resistin mRNA in cells treated with insulin alone was faster than that with actinomycin D. CONCLUSION/INTERPRETATION: Insulin downregulates resistin mRNA via PI 3-kinase, ERK or p38 MAP-kinase independent pathways in 3T3-L1 adipocytes. The downregulation mechanism of resistin mRNA by insulin would be an indirect event through the synthesis of novel protein(s) that could accelerate the degradation of resistin mRNA. PMID- 12627323 TI - Characterisation of new KATP-channel mutations associated with congenital hyperinsulinism in the Finnish population. AB - AIMS/HYPOTHESIS: ATP-sensitive potassium (K(ATP)) channels are crucial for the regulation of insulin secretion from pancreatic beta cells and mutations in either the Kir6.2 or SUR1 subunit of this channel can cause congenital hyperinsulinism (CHI). The aim of this study was to analyse the functional consequences of four CHI mutations (A1457T, V1550D and L1551V in SUR1, and K67N in Kir6.2) recently identified in the Finnish population. METHODS: Wild type or mutant Kir6.2 and SUR1 subunits were coexpressed in Xenopus oocytes. The functional properties of the channels were examined by measuring currents in intact oocytes or giant inside-out membrane patches. Surface expression was measured by enzyme-linked immunosorbance assay, using HA-epitope-tagged subunits. RESULTS: Two mutations (A1457T and V1550D) prevented trafficking of the channel to the plasma membrane. The L1551V mutation reduced surface expression 40-fold, and caused loss of MgADP and diazoxide activation. Both these factors will contribute to the lack of K(ATP) current activation observed in response to metabolic inhibition in intact oocytes. The L1551V mutation also increased the channel open probability, thereby producing a reduction in ATP-sensitivity (from 10 micro mol/l to 120 micro mol/l). The fourth mutation (K67N mutation in Kir6.2) did not affect surface expression nor alter the properties of K(ATP) channels in excised patches, but resulted in a reduced K(ATP) current amplitude in intact cells on metabolic inhibition, through an unidentified mechanism. CONCLUSION/INTERPRETATION: The four CHI mutations disrupted K(ATP) channel activity by different mechanisms. Our results are discussed in relation to the CHI phenotype observed in patients with these mutations. PMID- 12627324 TI - AICA-riboside induces apoptosis of pancreatic beta cells through stimulation of AMP-activated protein kinase. AB - AIMS/HYPOTHESIS: Prolonged exposure of beta cells to low glucose concentrations triggers their apoptosis and is known to activate AMP-activated protein kinase (AMPK) in beta cell lines. We examined whether prolonged activation of AMPK can trigger apoptosis in rodent beta cells. METHODS: Primary beta cells were FACS purified from rats, and from wild-type and AMPK(alpha2)-deficient mice. AMPK activation in beta cells was induced by the adenosine analog AICA-riboside and detected by immunoblotting using a phosphospecific antibody. Apoptosis of rodent beta cells was monitored by FACS analysis of beta cell DNA content, by direct counting of apoptotic cells using fluorescence microscopy, or by measurement of their caspase-3 activity. RESULTS: Dose-dependent and time-dependent apoptosis of the cells, concommittant with an activation of caspase-3, were suppressed by the caspase inhibitors zVAD-fmk and zDEVD-fmk. Apoptosis induction by AICA-riboside was also prevented by adding the MAPK-inhibitor SB203580 which blocked the AICA riboside-induced phosphorylation of AMPK. Beta cells isolated from AMPK-(alpha2) deficient mice were resistant against AICA-riboside induced apoptosis. CONCLUSION/INTERPRETATION: Sustained activation of AMPK by AICA-riboside can trigger a caspase-dependent apoptosis of pancreatic beta cells. PMID- 12627325 TI - IL-1beta and IFN-gamma induce the expression of diverse chemokines and IL-15 in human and rat pancreatic islet cells, and in islets from pre-diabetic NOD mice. AB - AIMS/HYPOTHESIS: Cytokines and chemokines are important mediators of immune responses due to their ability to recruit and activate leukocytes. Using microarray analysis we observed that rat beta cells exposed to IL-1beta and IFN gamma have increased mRNA levels of chemokines and IL-15. The aim of this study was to characterize the expression of IP-10, MIP-3alpha, fractalkine and IL-15 in rat beta cells, human pancreatic islets, and in islets isolated from NOD mice, both during the pre-diabetic period and following islet transplantation. METHODS: FACS-purified rat beta cells and human islets were cultured with IL-1beta, IFN gamma and/or TNF-alpha. Islets were isolated from NOD or BALB/c mice at different ages. For syngeneic islet transplantation, 2- or 3-week-old NOD islets were grafted under the kidney capsule of spontaneously diabetic NOD recipients. Chemokine and IL-15 mRNA expression and protein release were evaluated, respectively, by RT-PCR and ELISA. RESULTS: Human islets and rat beta cells express IP-10, MIP-3alpha, fractalkine and IL-15 mRNAs upon exposure to cytokines. The expression of IL-15, IP-10 and fractalkine is regulated by IFN gamma, while the expression of MIP-3alpha is IL-1beta-dependent. Moreover, cytokines induced IL-15, IP-10, Mig, I-TAC and MIP-3alpha protein accumulation in culture medium from human islets. In vivo, there was an age-related increase in IL-15, IP-10 and MIP-3alpha expression in islets isolated from NOD mice. Following syngeneic islet transplantation, increased expression of IL-1beta, IFN gamma, fractalkine, IP-10, MCP-1 and MIP-3alpha mRNAs were observed in the grafts. CONCLUSION/INTERPRETATION: Cytokine-exposed islets or beta cells express chemokines and IL-15. This could contribute to the recruitment and activation of mononuclear cells and development of insulitis in early Type 1 diabetes and during graft destruction. PMID- 12627326 TI - Alterations of left ventricular function in women with insulin-dependent diabetes mellitus during pregnancy. AB - AIMS/HYPOTHESIS: During pregnancy, eminent cardiovascular changes occur. The aim of the following study was to investigate the course of haemodynamic parameters under the increased volume load during pregnancy and delivery in women with insulin-dependent diabetes mellitus. METHODS: We examined 51 pregnant diabetic women and 51 healthy pregnant women. The control group consisted of 51 healthy non-pregnant women. In all women, left ventricular mass and fractional shortening were calculated. To evaluate left ventricular diastolic function, mitral inflow and pulmonary venous flow profiles were analysed. RESULTS: During pregnancy left ventricular mass increased, fractional shortening decreased and diastolic dysfunction was found. While the healthy pregnant women developed signs of disturbed relaxation during pregnancy, pregnant diabetic women showed signs of a disturbed relaxation at the beginning of gestation. Of the pregnant diabetic women, 29 developed a restrictive filling pattern at the 24th week of gestation. The remaining 22 diabetic women had a comparable restrictive filling pattern only during vaginal delivery. In 10 of the 29 pregnant diabetic women dangerous complications were documented, while there were no complications in the healthy pregnant women and the other 22 diabetic pregnant women. CONCLUSION/INTERPRETATION: In healthy women pregnancy results in a reversible physiologic left ventricular hypertrophy, a disturbed relaxation pattern and a temporary decrease of left ventricular systolic function. In contrast, pregnant diabetic women showed a delayed relaxation at the beginning of pregnancy and developed a restrictive filling pattern. The early development of a restrictive filling pattern could indicate complications during delivery in pregnant diabetic women. PMID- 12627328 TI - Pigment epithelium-derived factor prevents advanced glycation end products induced monocyte chemoattractant protein-1 production in microvascular endothelial cells by suppressing intracellular reactive oxygen species generation. AB - AIMS/HYPOTHESIS: Monocytes and macrophages accumulate in the lesion of the diabetic retina, which are most likely involved in the progression of diabetic retinopathy. The levels of monocyte chemoattractant protein-1 (MCP-1) in vitreous fluids were associated with the severity of proliferative diabetic retinopathy. Recently, pigment epithelium-derived factor has been shown to be involved in the pathogenesis of proliferative diabetic retinopathy. However, a role of pigment epithelium-derived factor in monocyte recruitments in diabetic retinopathy remains to be elucidated. In this study, we investigated effects of purified pigment epithelium-derived factor on AGE-induced reactive oxygen species generation, MCP-1 mRNA up-regulation and protein production in human cultured microvascular endothelial cells. METHODS: The intracellular formation of reactive oxygen species was measured using the fluorescent probe CM-H(2)DCFDA. MCP-1 gene expression was analysed in quantitative reverse transcription-polymerase chain reaction. Monocyte chemoattractant protein-1 production by microvascular endothelial cells was measured with an ELISA system. RESULTS: AGE increased intracellular reactive oxygen species generation in microvascular endothelial cells. Pigment epithelium-derived factor inhibited the AGE-induced reactive oxygen species generation in a dose-dependent manner. An anti-oxidant, N acetylcysteine, or pigment epithelium-derived factor completely prevented the AGE induced up-regulation of MCP-1 mRNA contents as well as protein production in microvascular endothelial cells. CONCLUSIONS/INTERPRETATIONS: Pigment epithelium derived factor inhibits the AGE-induced reactive oxygen species generation and the subsequent increase in MCP-1 production in microvascular endothelial cells. Our study suggests that substitution of pigment epithelium-derived factor could prevent the progression of diabetic retinopathy by attenuating the deleterious effects of AGE. PMID- 12627327 TI - Effect of high concentrations of glucose on differentiation of rat trophoblast cells in vitro. AB - AIMS/HYPOTHESIS: Previous studies have shown that diabetic placentas are characterized by structural and biochemical anomalies, including defects in the differentiation of trophoblasts. In this study, the Rcho-1 cell line was used to investigate the impact of high glucose concentrations on different markers of differentiation of rat trophoblast cells in giant cells (endoreduplication, invasive phenotype and endocrine phenotype). MATERIALS: Rcho-1 cells were incubated for 12 days in medium supplemented with different concentrations of glucose and 10% horse serum to optimize differentiation. The cells were examined for the proportion of nuclei showing signs of apoptosis. The effect of high glucose was investigated on the endoreduplication process, on invasive phenotype (secretion of gelatinase B) and on endocrine phenotype (expression of placental lactogen I (PL-I) and II (PL-II) and progesterone secretion). RESULTS: Apoptosis was not induced by high glucose in Rcho-1. The number of cells was higher in the cultures exposed to high glucose (p<0.05) and their nuclei contained more DNA compared with control cells (p<0.001), while their nuclear size was smaller (p<0.001). Gelatinase B secretion increased during differentiation but no difference was found when gelatinase B secretion from trophoblasts exposed to high glucose was compared with the control cells. Rcho-1 cell cultures showed an increase in PL-I and PL-II mRNA expressions during differentiation and which was not affected by high glucose. Progesterone secretion increased during differentiation in control cultures. However, this increase was abolished when trophoblasts were cultured in high glucose. CONCLUSIONS/INTERPRETATION: Our data suggest that high glucose influences the endoreduplication process and the steroidogenesis during differentiation of rattrophoblasts. PMID- 12627329 TI - Rosiglitazone and pulmonary oedema: an acute dose-dependent effect on human endothelial cell permeability. AB - AIMS/HYPOTHESIS: Peripheral and pulmonary oedema has emerged as the most common drug-related side effect of rosiglitazone in clinical practice, but the underlying mechanisms are not clear. Fluid retention and changes in vascular tone could contribute to oedema formation, but the interpretation of clinical and in vivo studies is particularly difficult and the direct effects of thiazolidinediones on endothelial barrier function have not been previously reported. METHODS: Human pulmonary artery endothelial cells were seeded and grown on 0.4 microm collagen-coated filters to form a tight monolayer (transendothelial electrical resistance 9-11 ohms.cm(-2) after 2-3 days). Transendothelial albumin flux (expressed as the percentage clearance of albumin relative to control) was measured using Evans blue-labelled albumin after exposure to rosiglitazone 1-100 micromol/l for 1 h to 48 h, and after removal of drug from the monolayer. RESULTS: Incubation of pulmonary artery endothelial cells with rosiglitazone for 4 h produced immediate concentration-dependent increases in transendothelial albumin flux: e.g., relative to control (100%), 113%+/-13% (1 micromol/l), 215%+/ 37% (10 micromol/l, p=0.01) and 461%+/-96% (100 micromol/l, p=0.002) (n=12). There was no effect after 1 h. The acute hyperpermeability response to rosiglitazone, maximal after 4 h, was fully reversible after washing the monolayer. After incubation for 24 to 48 h the effect of rosiglitazone on pulmonary endothelial permeability tended to subside: e.g., 210%+/-59% (24 h) for rosiglitazone 100 micromol/l (p=0.06). CONCLUSION/INTERPRETATION: Exposure to high-therapeutic concentrations of rosiglitazone causes a reversible fourfold increase in pulmonary endothelial permeability which could be clinically relevant especially at higher doses and at times of peak plasma drug concentration. PMID- 12627330 TI - Genetic epidemiology of MODY in the Czech republic: new mutations in the MODY genes HNF-4alpha, GCK and HNF-1alpha. AB - AIMS/HYPOTHESIS: The aim of this study was to examine the prevalence and nature of mutations in HNF4alpha/MODY1, GCK/MODY2 and HNF-1alpha/MODY3 genes in Czech subjects with clinical diagnosis of MODY. METHODS: We studied 61 unrelated index probands of Czech origin (28 males, 33 females) with a clinical diagnosis of MODY and 202 family members. The mean age of probands was 22.7+/-12.0 years (range, 6 62) and the mean age at the first recognition of hyperglycaemia was 14.7+/-6.0 years (range, 1-25). The promotor and coding regions inclusive intron exon boundaries of the HNF-4alpha, GCK and HNF-1alpha genes were examined by PCR-dHPLC (HNF-1alpha and GCK) and direct sequencing. RESULTS: We identified 20 different mutations in the HNF-4alpha, GCK and HNF-1alpha in 29 families (48% of all families studied), giving a relative prevalence of 5% of MODY1, 31% of MODY2 and 11.5% of MODY3 among the Czech kindred with MODY. Three of 3, 10 of 11 and 1 of 6 of the mutations identified in HNF-4alpha, GCK and HNF-1alpha respectively, were new. CONCLUSION/INTERPRETATION: Of the families 48% carried mutations in the MODY1-3 genes and of the identified mutations 70% were new. In 52% of Czech families with clinical characteristics of MODY, no mutations were found in the analysed genes. This finding shows that the majority of MODY mutations in a central European population are local and that other MODY genes could be responsible for autosomal dominant transmission of diabetes mellitus. PMID- 12627331 TI - Heteroplasmic ratio of the A3243G mitochondrial DNA mutation in single pancreatic beta cells. AB - AIMS/HYPOTHESIS: To examine whether there is a high content of mutated mitochondrial DNA in individual pancreatic beta cells from a patient with the A3243G mitochondrial DNA mutation. METHODS: Tissues were available from a patient with diabetes and the A3243G mutation including pancreatic tissue. We quantified the amount of mutated mitochondrial DNA in tissue homogenates and single pancreatic beta cells using hot last cycle PCR. RESULTS: The percentage ratio of mutated to wild-type mtDNA was high in tissues such as muscle and brain (>60%), but surprisingly low in both pancreatic islets and in individual beta cells from these islets. The islets were smaller in the patient than in control subjects in keeping with a decreased beta-cell mass. CONCLUSIONS/INTERPRETATION: These observations suggest that either the beta cells show increased sensitivity to the effects mtDNA mutations on respiratory chain function, and/or cells with a high mutant load are preferentially removed leading to a progressive decrease in the islet beta-cell mass. PMID- 12627332 TI - Free fatty acids (FFA) and endothelial dysfunction; role of increased oxidative stress and inflammation. --to: Steinberg et al. (2002) Vascular function, insulin resistance and fatty acids. PMID- 12627333 TI - --to: Gale EAM (2002) A missing link in the hygiene hypothesis? Diabetologia 45:588-592. PMID- 12627336 TI - --to: Hales CN, Barker DJP (1992) Type 2 (non-insulin-dependent) diabetes mellitus: the thrifty phenotype hypothesis. Diabetologia 35:595-601. PMID- 12627334 TI - --to: M. Koopmanschap: Coping with Type 2 diabetes: the patient's perspective. Diabetologia 45:S18-S22. PMID- 12627337 TI - [Cholangioscopy after successful treatment of complicated choledocholithiasis. Is stone free really stone free?]. AB - BACKGROUND AND OBJECTIVE: After successful percutaneous or endoscopic therapy of complicated choledocholithiasis (requiring more than one therapeutic intervention or lithotripsy), radiological visualization of the bile duct is the standard to determine if any stone fragments are left. It is unknown how often stone fragments, which might be the cause for another period of symptomatic choledocholithiasis, are missed. PATIENTS AND METHODS: We performed cholangioscopy in 31 consecutive patients (age 42 - 85; 14 male, 17 female) with complicated choledocholithiasis after successful therapy when there were no stone fragments left radiographically. RESULTS: Cholangioscopy revealed retained stone fragments in four female patients. Two of these had a benign stricture of the common bile duct. CONCLUSIONS: Cholangioscopy after successful endoscopic or percutaneous therapy of complicated choledocholithiasis seems to be useful, because in bile ducts radiographically free of stones, cholangioscopy detects stone fragments in some patients. Further studies are needed to demonstrate if cholangioscopy can reduce the rate of symptomatic relapse after treatment of complicated choledocholithiasis. PMID- 12627338 TI - [Control of systolic blood pressure in the VALUE trial after 12 months]. PMID- 12627339 TI - [Exudative erythema multiforme with transition to a toxic epidermal necrolysis after taking aceclofenac (Beofenac)]. AB - HISTORY AND CLINICAL FINDINGS: A 75-year-old woman took aceclofenac (100 mg/day) for 15 days for long-standing arthritis. Starting with erythema of the face she developed multiple target lesions with central bullae on her neck, chest, back, including the palmoplantar region. In several areas the lesions became confluent and showed wide detachment. The mucous membranes of the eyes, mouth and genitalia also showed marked inflammation, most prominent were large erosions in the mouth. The clinical course culminated in an overall skin loss of about 15 % of the total body surface area. DIAGNOSIS, TREATMENT AND COURSE: Treatment with topical and systemic corticosteroids as well as antiseptics resulted in complete clearing of the cutaneous lesions within 4 weeks. Various analgetics, particularly the closely related diclofenac, were tolerated without any side-effects after oral intake. Re-exposition with aceclofenac was rejected due to safety reasons. CONCLUSION: This is the first report of a potentially life-threatening adverse cutaneous drug reaction after taking with aceclofenac. PMID- 12627340 TI - [Hyperthyroidism--case report]. PMID- 12627341 TI - [Diagnosis of hyperthyroidism]. PMID- 12627342 TI - [Therapy of hyperthyroidism]. PMID- 12627345 TI - [The glutathione system in the human gastrointestinal tract. Significance and influence of clinical factors]. PMID- 12627346 TI - [Claims for compensation by physicians using alternative treatment. Verdict from the Stuttgart Provincial High Court and Court of Appeals from 4-16-2002 -1(14) U 71/2001]. PMID- 12627347 TI - [What is to be made of the therapy of borreliosis with cholestyramine and pioglitazone?]. PMID- 12627348 TI - [Primary prevention of sudden death--afraid of the truth]. PMID- 12627349 TI - [Clinical ethics consultation--a new field for medical ethics?]. PMID- 12627350 TI - Hajj-associated outbreak strain of Neisseria meningitidis serogroup W135: estimates of the attack rate in a defined population and the risk of invasive disease developing in carriers. AB - An outbreak of disease due to Neisseria meningitidis serogroup W135 (W135) occurred in 2000 and 2001 among pilgrims returning from the annual Islamic pilgrimage to Saudi Arabia (the Hajj) and in their contacts. For the Hajj in 2000, the attack rate of W135 disease was 25 cases per 100,000 pilgrims. After the introduction of quadrivalent meningococcal vaccine for the Hajj in 2001, no pilgrim developed W135 disease. The estimated attack rates for household contacts of returning pilgrims were 18 cases and 28 cases per 100,000 contacts for the years 2000 and 2001, respectively. On the basis of rates of transmission of W135 carriage and national epidemiological data, the risk that an unvaccinated household contact who had acquired W135 carriage would develop invasive meningococcal disease was estimated to be 1 case per 70 acquisitions. Public health policies to protect household contacts of Hajj pilgrims need to be implemented. PMID- 12627351 TI - Epidemiology of meningococcal disease in light of recent Hajj-associated outbreaks. PMID- 12627352 TI - Prevalence and clinical correlates of YMDD variants during lamivudine therapy for patients with chronic hepatitis B. AB - YMDD variants of hepatitis B virus (HBV) emerge in some patients with chronic hepatitis B who receive lamivudine. YMDD variants were examined in 794 patients in 4 controlled studies of 1 year's duration. The long-term effects of YMDD variants were examined in a subset of patients treated up to 4 years. YMDD variants were detected by polymerase chain reaction (PCR) and restriction fragment-length polymorphism assays. After 1 year, YMDD variants were detected in 81 (24%) of 335 patients. In these patients, the median serum HBV DNA concentration at 1 year was <20% of the baseline level, and serum alanine transaminase (ALT) levels and liver histologic findings had significantly improved. In patients with YMDD variants who were treated for up to 4 years, median HBV DNA and ALT levels showed improvements. Sex, baseline body mass index, and HBV DNA level were associated with emergence of YMDD variants. Patients with YMDD variants losing clinical response with a significant increase in the HBV DNA and ALT levels may require additional therapy. PMID- 12627353 TI - Gram stain, culture, and histopathological examination findings for heart valves removed because of infective endocarditis. AB - Retrospective chart review was undertaken for 480 patients who underwent a total of 506 valve replacements or repair procedures for infective endocarditis. The influence of preoperative antimicrobial treatment on culture, Gram stain, and histopathological examination findings for resected valve specimens was examined. When valves were removed before the end of treatment, organisms were seen on the Gram stain of ground valve material performed in the microbiology laboratory and on Gram-stained histopathological sections in 231 (81%) of 285 and 140 (67%) of 208 specimens, respectively (P=.0007). Gram-positive cocci were either cultured from or observed in excised valve tissue in 42 (67%) of 63 episodes involving negative preoperative blood cultures. Positive Gram stain results for microbiological specimens should be reintroduced into the definite pathological criteria for infective endocarditis. When deciding on how long to continue antimicrobial therapy after valve replacement for endocarditis, valve culture results should be the only laboratory finding taken into account, because it takes months for dead bacteria to be removed from sterile vegetations. PMID- 12627354 TI - Oculo-respiratory syndrome: a new influenza vaccine-associated adverse event? AB - During the 2000-2001 influenza immunization campaign in Canada, a new adverse event, oculo-respiratory syndrome (ORS), was noted in association with administration of vaccine supplied by one manufacturer. The original case definition for ORS specified bilateral conjunctivitis, facial edema, or respiratory symptoms beginning 2-24 h after influenza vaccination and resolving within 48 h after onset. To characterize the spectrum, severity, and impact of ORS, we contacted persons who had reported any influenza vaccine-associated adverse event in British Columbia, Canada, during the 2000-2001 vaccination campaign. With use of a standardized telephone interview, we collected information from 609 (79%) of 769 eligible persons. Thirteen percent of ORS affected persons reported onset 48 h, and 42% considered the symptoms to be severe. The surveillance case definition for ORS for 2001-2002 was revised to include onset .1). AC-related infection occurred in 3 patients in the SBP group and in 7 patients in the control group (P>.1). Five episodes of AC-related bloodstream infection were diagnosed (1.5 cases per 1000 catheter-days). Use of SBPs did not prevent AC colonization or infection. The incidence of AC-related infectious complications was comparable to the incidence of CVC-related infection reported in the literature. PMID- 12627357 TI - In search of encephalitis etiologies: diagnostic challenges in the California Encephalitis Project, 1998-2000. AB - The California Encephalitis Project was initiated in June 1998 to identify the causes and characterize the clinical and epidemiologic features of encephalitis in California. Testing for >or=13 agents, including herpesviruses, enteroviruses, arboviruses, Bartonella species, Chlamydia species, and Mycoplasma pneumoniae, was performed at the Viral and Rickettsial Disease Laboratory (Richmond, California). Epidemiologic and clinical information collected for each case guided further testing. From June 1998 through December 2000, 334 patients who met our case definition of encephalitis were enrolled. A confirmed or probable viral agent of encephalitis was found in 31 cases (9%), a bacterial agent was found in 9 cases (3%), and a parasitic agent was found in 2 cases (1%). A possible etiology was identified in 41 cases (12%). A noninfectious etiology was identified in 32 cases (10%), and a nonencephalitis infection was identified in 11 (3%). Despite extensive testing and evaluation, the etiology of 208 cases (62%) remained unexplained. PMID- 12627359 TI - Randomized comparison of oral valacyclovir and intravenous ganciclovir for prevention of cytomegalovirus disease after allogeneic bone marrow transplantation. AB - In this multicenter, randomized study, cytomegalovirus (CMV)-seropositive patients who received an allogeneic bone marrow transplant were provided high dose intravenous acyclovir (500 mg/m(2) q8h) from the day of transplantation until engraftment. The patients were then randomly assigned to receive either oral valacyclovir, 2 g q.i.d. (n=83), or intravenous ganciclovir, 5 mg/kg q12h for 1 week, then 6 mg/kg once daily for 5 days per week (n=85), until day 100 after transplantation. CMV infection occurred in 12% of the patients who received valacyclovir and in 19% of the patients who received ganciclovir (hazard ratio [HR], 1.042; 95% confidence interval [CI], 0.391-2.778; P=.934). CMV disease developed in only 2 patients who received valacyclovir and in 1 patient who received ganciclovir (HR, 1.943; 95% CI, 0.176-21.44; P=.588). Oral valacyclovir can be an effective alternative to intravenous ganciclovir for prophylaxis of CMV disease after bone marrow transplantation. PMID- 12627360 TI - Risk factors for postcraniotomy surgical site infection after 1,3-bis (2 chloroethyl)-1-nitrosourea (Gliadel) wafer placement. AB - Gliadel wafers (1,3-bis [2-chloroethyl]-1-nitrosourea; Guilford Pharmaceuticals) are approved for the treatment of malignant gliomas; however, the incidence of and risk factors associated with infection with respect to this new technology are unknown. We identified 32 patients who received Gliadel wafers from December 1996 through October 1999. Nine patients (28%) developed >or=1 surgical site infection (SSI), which included 4 cases of brain abscess. All 3 patients who received vancomycin for surgical prophylaxis developed an SSI. In addition, multivariable analysis revealed an association between infection and a clinical diagnosis of depression. The National Nosocomial Infection Surveillance Surgical Site Index did not predict the onset of SSI after Gliadel wafer implantation. Patients who received a Gliadel wafer had a higher incidence of infection than previously has been reported, and additional studies are required to better quantify this risk and describe the epidemiology of such infections. PMID- 12627361 TI - Progressive vaccinia. AB - The resumption of smallpox vaccination for health care workers and other first responders has raised concern about the occurrence of complications in people with immunodeficiency disorders, including those infected with human immunodeficiency virus. During the era of universal vaccination, roughly 1 person per million vaccinees in the general population developed progressive vaccinia, which is characterized by the relentless outward spread of infection from the vaccination site and eventual dissemination to other areas on the body. Review of 56 cases reported in the English-language medical literature from 1893 through 1997 indicates that the condition occurred only in persons with severe cell mediated immunodeficiency. Progressive vaccinia was found to be lethal in infants who completely lacked cellular immune function, but infection resolved in many adults with acquired immunodeficiency. Almost all cases were treated with vaccinia immune globulin, but its efficacy has never been tested in a placebo controlled trial. Further research is needed to develop effective forms of therapy. PMID- 12627362 TI - Safety of probiotics that contain lactobacilli or bifidobacteria. AB - Lactobacilli and bifidobacteria are extremely rare causes of infection in humans, as are probiotics based on these organisms. This lack of pathogenicity extends across all age groups and to immunocompromised individuals. Strains used for new probiotics should be chosen from the commensal flora of humans and should not carry intrinsic resistance to antibiotics that would prevent treatment of a rare probiotic infection. Vigilance regarding the detection of possible rare cases of infection due to probiotics should be maintained, and isolates should be sent to reference centers for molecular characterization and confirmation. PMID- 12627363 TI - The role of clonality in the global spread of fluoroquinolone-resistant bacteria. AB - The molecular epidemiology of fluoroquinolone resistance in a number of bacterial pathogens suggests that the persistence and spread of resistance is associated with a small number of highly successful bacterial clones. The recent description of fluoroquinolone-resistant strains among global clones of pneumococci raises the likelihood that the percentage of pneumococci resistant to fluoroquinolones will increase as this class of agent is increasingly used for the management of respiratory tract infection in adults. Although these clones remain fluoroquinolone susceptible in children, their widespread distribution argues against the extension of fluoroquinolone use to the management of respiratory tract infection in children. PMID- 12627364 TI - Smallpox vaccination and the patient with an organ transplant. AB - Organ transplant recipients are vulnerable to a variety of infections because of the immunosuppressed state required to prevent organ rejection. We review the recommendations of the Centers for Disease Control and Prevention (Atlanta, Georgia) for smallpox vaccination and the possible complications of vaccination in the population with organ transplants. The risk of these complications is presumably dependent on the extent of deficient cell-mediated immunity. PMID- 12627365 TI - The changing epidemiology of cryptococcosis: an update from population-based active surveillance in 2 large metropolitan areas, 1992-2000. AB - To examine trends in the incidence and epidemiology of cryptococcosis, active, population-based surveillance was conducted during 1992-2000 in 2 areas of the United States (the Atlanta, Georgia, and Houston, Texas, metropolitan areas; combined population, 7.4 million). A total of 1491 incident cases were detected, of which 1322 (89%) occurred in HIV-infected persons. The annual incidence of cryptococcosis per 1000 persons with AIDS decreased significantly during the study period, from 66 in 1992 to 7 in 2000 in the Atlanta area, and from 24 in 1993 to 2 in 1994 in the Houston area. Poisson regression analysis revealed that African American persons with AIDS were more likely than white persons with AIDS to develop disease. Less than one-third of all HIV-infected persons with cryptococcosis were receiving antiretroviral therapy before diagnosis. Our findings suggest that HIV-infected persons who continue to develop cryptococcosis in the era of highly active antiretroviral therapy (HAART) in the United States are those with limited access to health care. More efforts are needed to expand the availability of HAART and routine HIV care services to these persons. PMID- 12627366 TI - Human immunodeficiency virus type 1-related lipoatrophy and lipohypertrophy are associated with serum concentrations of leptin. AB - The relationship between the adipocyte-derived hormone leptin, insulin resistance, and fat redistribution in patients with human immunodeficiency virus (HIV) infection has not been established. We classified a cohort of HIV type 1 (HIV-1)-infected patients with >or=6 months of antiretroviral exposure as having no lipodystrophy (51 patients [43% of the cohort]), lipoatrophy (23 patients [19% of the cohort]), mixed lipodystrophy (29 patients [24% of the cohort]), or lipohypertrophy (17 patients [14% of the cohort]), on the basis of physical examination, anthropometric measurements, and the findings of dual-emission x-ray absorptiometry and computed tomography. Measurements of insulin resistance were higher for patients with each category of lipodystrophy, compared with those observed for patients with no lipodystrophy (P<.001). Mean leptin levels (+/- standard deviation) were lowest in patients with lipoatrophy (1.76+/-1.20 ng/mL), highest in patients with lipohypertrophy (9.10+/-6.86 ng/mL), and significantly different from those in patients without lipodystrophy (3.14+/-2.30 ng/mL; both P<.01). In this cohort of antiretroviral-experienced HIV-infected patients, a low serum level of leptin was independently associated with insulin resistance in patients with lipoatrophy, after controlling for total and regional body fat. PMID- 12627367 TI - Electronic human immunodeficiency virus (HIV) clinical reminder system improves adherence to practice guidelines among the University of Washington HIV Study Cohort. AB - We conducted a prospective study of an electronic clinical reminder system in an academic medical center-based human immunodeficiency virus (HIV) specialty clinic. Published performance indicators were used to examine adherence to HIV practice guidelines before and after its implementation for 1204 patients. More than 90% of patients received CD4 cell count and HIV type 1 (HIV-1) RNA level monitoring every 3-6 months during both time periods, and approximately 80% of patients with a CD4 cell count nadir of <350 cells/mm(3) received highly active antiretroviral therapy. Patients were significantly more likely to receive prophylaxis against Mycobacterium avium complex (hazard ratio, 3.84; 95% confidence interval [CI], 1.58-9.31; P=.003), to undergo annual cervical carcinoma screening (OR, 2.09; 95% CI, 1.04-4.16; P=.04), and to undergo serological screening for Toxoplasma gondii (odds ratio [OR], 1.86; 95% CI, 1.05 3.27; P=.03) and syphilis infection (OR, 3.71; 95% CI, 2.37-5.81; P<.0001). HIV clinical reminders delivered at the time that HIV care is provided were associated with more timely initiation of recommended practices. PMID- 12627368 TI - Initiation of highly active antiretroviral therapy at CD4+ T lymphocyte counts of >350 cells/mm3: disease progression, treatment durability, and drug toxicity. AB - We compared clinical disease progression in 159 human immunodeficiency virus (HIV)-infected persons for whom highly active antiretroviral therapy (HAART) was initiated when they had CD4(+) T lymphocyte counts of 350-499 cells/mm(3) with progression in 174 HIV-infected patients for whom it was not. Disease progression did not differ between the 2 groups (P=.21, log-rank test). Fifty-three percent of the 159 treated patients had HIV type 1 RNA levels of >400 copies/mL at the most recent evaluation, and 41% had experienced adverse drug reactions necessitating a change in regimen. These findings support the recommendation that HAART not be initiated for patients with CD4(+) cell counts of >350 cells/mm(3). PMID- 12627369 TI - Longitudinal study of herpesviruses in kidney transplant recipients in Cuba. AB - We compared a multiplex polymerase chain reaction assay and a shell vial assay for the detection of herpesviruses infection in 13 Cuban patients who had received kidney transplants. Cytomegalovirus and human herpesvirus 6 were detected in these patients. PMID- 12627370 TI - Serum procalcitonin levels and empirical antibiotic treatment of patients with community-acquired febrile syndromes. PMID- 12627371 TI - The future diagnostic role of procalcitonin levels: the need for improved sensitivity. PMID- 12627372 TI - Procalcitonin: what should be its role in the clinical management of febrile patients admitted to the hospital? PMID- 12627373 TI - High serum procalcitonin levels do not predict bacteremia in adult patients with acute fever. PMID- 12627377 TI - Evaluation of nonionic and zwitterionic detergents as membrane protein solubilizers in two-dimensional electrophoresis. AB - The solubilizing power of various nonionic and zwitterionic detergents as membrane protein solubilizers for two-dimensional electrophoresis was investigated. Human red blood cell ghosts and Arabidopsis thaliana leaf membrane proteins were used as model systems. Efficient detergents could be found in each class, i.e. with oligooxyethylene, sugar or sulfobetaine polar heads. Among the commercially available nonionic detergents, dodecyl maltoside and decaethylene glycol mono hexadecyl ether proved most efficient. They complement the more classical sulfobetaine detergents to widen the scope of useful detergents for the solubilization of membrane proteins in proteomics. PMID- 12627376 TI - A simple affinity spin tube filter method for removing high-abundant common proteins or enriching low-abundant biomarkers for serum proteomic analysis. AB - Although it is possible to identify new proteins from crude cell extracts using proteomics technology, it is often difficult to elucidate low-abundant biomarkers in the presence of a large amount of high-abundant proteins in serum. We have developed a simple and rapid method using an affinity spin tube filter to remove high-abundant common proteins and enrich the low-abundant biomarkers. The affinity spin tube filter contains protein G, coupled with antibodies against either high-abundant proteins or specific proteins of interest. After incubating with serum, the flow-through or the elute was collected and analyzed by two dimensional gel electrophoresis. By using this affinity spin tube filter, the possibilities of identifying new biomarkers are shown. This technique could be used for large-scale sample preparation for high-throughput proteomic analysis. PMID- 12627378 TI - Antibody microarrays: an evaluation of production parameters. AB - Antibody microarrays could have an enormous impact on the functional analysis of cellular activity and regulation, especially at the level of protein expression and protein-protein interaction, and might become an invaluable tool in disease diagnostics. The array surface is bound to have a tremendous influence on the findings from such studies. Apart from the basic issue of how to attach antibodies optimally without affecting their function, it is also important that the cognate antigens, applied within a complex protein mixture, all bind to the arrayed antibodies irrespective of their enormous variety in structure. In this study, various factors in the production of antibody microarrays on glass support were analysed: the modification of the glass surface; kind and length of cross linkers; composition and pH of the spotting buffer; blocking reagents; antibody concentration and storage procedures, in order to evaluate their effect on array performance. Altogether, data from more than 700 individual array experiments were taken into account. In addition to home-made slides, commercially available systems were also included in the analysis. PMID- 12627379 TI - A new method to improve sensitivity and resolution in matrix-assisted laser desorption/ionization time of flight mass spectrometry. AB - High detection sensitivity and resolution are two critical parameters for recording good peptide mass fingerprints (PMF) of low abundance proteins. This paper reports a mass spectrometry (MS) sample preparation technique that could improve sensitivity and resolution. By coating the MS steel target with a thin layer of pentadecafluorooctamido propyltrimethoxysilane, which was both polar and nonpolar solvent repellent, the transferred sample droplets on its surface were significantly smaller. As a result, the analyte of the peptide mixture became more concentrated and homogeneous, which helped to improve the sensitivity. The advantages of a modified MS target were documented by mass spectra improvement of attomole level standard peptides and silver-stained proteins from polyacrylamide gels. The mass signal of angiotensin II at 100 attomole was difficult to record on the conventional support, whereas it was easily detected on the modified one. The PMF of cytochrome C was also better recorded on the modified support, in terms of both signal-to-noise ratio and the number of detected peptides. When silver-stained proteins from two-dimensional electrophoresis gels were analyzed, in most cases more satisfactory peptide mass spectra were obtained from the modified support. Searching protein databases with more mass data from the improved PMFs, several unknown proteins were successfully identified. PMID- 12627380 TI - The rapid detection of low molecular mass proteins differentially expressed under biological stress for four Helicobacter spp. using ProteinChip technology. AB - Helicobacter pylori is one of the most prevalent human pathogens in the world and is the aetiological agent of gastritis, peptic ulcer disease and gastric malignancies. In addition H. pylori and other novel members of the genus are capable of successfully colonising the bile-rich niche of the upper intestine and are associated with a diverse range of intestinal pathologies. Surface-enhanced laser desorption/ionisation-time of flight mass spectrometry was used to analyse surface extracts from H. pylori, Helicobacter bilis, Helicobacter pullorum and "Helicobacter sp. flexispira" to characterise cell surface changes following bile stress. The system detected two distinct response patterns to bile stress on the cell surface of Helicobacter spp. in vitro. The first involved the increase under bile stress of peaks at 7.6 and 7.9 kDa for H. billis and H. pullorum, respectively. In contrast both "Helicobacter sp. flexispira" and a clinical isolate of H. pylori had similar response profiles to bile stress. Both strains had at least three low mass peaks decreased under bile stress and a single peak induced by bile stress. The present study has established the use of ProteinChip(R) technology to analyse helicobacter-related proteomics. Specifically this study has established that different patterns are generated in response to bile stress among various pathogenic Helicobacter spp. which may give insights into the ability of these strains to colonise different niches. PMID- 12627381 TI - Depletion of the highly abundant protein albumin from human plasma using the Gradiflow. AB - Analysis of complex protein samples by two-dimensional electrophoresis (2-DE) is often more difficult in the presence of a few predominant proteins. In plasma, proteins such as albumin mask proteins of lower abundance, as well as significantly limiting the amount of protein that can be loaded onto the immobilized pH gradient strip. In this paper the Gradiflow, a preparative electrophoresis system, has been used to deplete human plasma of the highly abundant protein albumin under native and denatured conditions. A three step protocol incorporating a charge separation to collect proteins with an isoelectric point greater than albumin and two size separations to isolate proteins larger and smaller than albumin, was used. When the albumin depleted fractions were analysed on pH 3-10 2-DE gels, proteins that were masked by albumin were revealed and proteins not seen in the unfractionated plasma sample were visualised. Matrix-assisted laser desorption/ionisation-time of flight mass spectrometry analysis confirmed the identification of the protein that lies beneath albumin to be C4B-binding protein alpha chain. The liquid fractions from the Gradiflow separations were also analysed by liquid chromatography-tandem mass spectrometry to confirm the proteins were separated according to their size and charge mobility in an electric field. PMID- 12627383 TI - The role of peptide deformylase in protein biosynthesis: a proteomic study. AB - Recently we investigated the influence of classical and emerging antibiotics on the proteome of Bacillus subtilis including in our studies actinonin, a potent novel inhibitor of peptide deformylase. The protein synthesis pattern under actinonin treatment changed so dramatically that a direct comparison to the control pattern was impossible. Dual channel imaging revealed that actinonin treatment caused the majority of newly synthesised proteins to accumulate in spots different from the ones usually observed, indicating a more acidic isoelectric point. Two strategies were used to investigate the nature of the charge shift. In the first place, protein patterns of a conditional peptide deformylase mutant under nonrepressing and repressing conditions were compared. Secondly, several protein pairs excised from two-dimensional (2-D) gels of the peptide deformylase mutant, exponentially growing untreated wild-type and the actinonin treated wild-type were investigated with matrix-assisted laser desorption/ionization and electrospray ionization (ESI) time of flight mass spectrometry (TOF MS) for the existence of N-terminal formylation. Under nonrepressing conditions the mutant protein pattern resembled that of the wild type. The loss of peptide deformylase activity under repressing conditions led to the same pI shift observed for actinonin treatment in the wild-type. Quadrupole TOF-MS on 11 protein pairs proved that the remaining N-terminal formyl residue was indeed responsible for the charge shift. Eight of these protein pairs were also present on 2-D gels of exponentially growing B. subtilis, where the more acidic, still formylated protein species represented the smaller parts. PMID- 12627384 TI - Wheat proteomics: relationship between fine chromosome deletion and protein expression. AB - To explore the relationship between fine chromosome deletion and protein expression in common wheat, the changes in protein composition of wheat seed proteome were investigated by using chromosome 1B. A momosomic alien chromosome addition line of common wheat was used to produce the fine deletion lines. Endosperm and embryo proteins were separated by two-dimensional gel electrophoresis (2-DE) and visualized by staining with Commassie Brilliant Blue, and gel images were analyzed with a computer assisted image analyzer. For the first time, fine gene locations of a few endosperm and embryo proteins were identified on the chromosome 1B. These proteins with their specific gene location on the chromosome can be used as protein markers in breeding programs for quality of wheat proteins. To identify wheat seed proteins and to understand their expression in relation to chromosome deletion, the feasibility of a new analytical approach based on isotope coded affinity tag labeling (ICAT) of peptides in tryptic digests followed by electrospray ionization mass spectrometry has been described. Simplification of the complex tryptic digest prior to mass spectral analysis was performed by treating the samples with light and heavy ICAT labeling reagents. A clear separation of peptide fragment containing the light and heavy reagents was achieved in mass spectral analysis. Out of the 14 peptides detected by mass fragment analysis of the euploid, four were down-regulated, nine up-regulated and one did not show any change due to the terminal deletion of chromosome 1B. Selected peptide fragments were subjected to tandem mass spectrometry analysis for sequence information and the resulting sequence information was submitted to databases for protein identification. Of the five proteins submitted, four were identified as alpha-amylase inhibitor, alpha amylase/subtilisin inhibitor precursor, proteasome subunit alpha-type 7 and 1,4 alpha-glucan-D-maltohydrolase. With this approach it is possible to identify wheat seed proteins and to understand their expression, which have been reported to be difficult by 2-DE due to cosynthesis of proteins by genes from three genomes, A, B and D. PMID- 12627382 TI - Proteomic profiling of proteins associated with urokinase plasminogen activator receptor in a colon cancer cell line using an antisense approach. AB - Expression of urokinase plasminogen activator (uPA) and its receptor (uPAR) strongly correlates with a malignant tumour cell phenotype. In the multistep process of metastasis, uPA binding to uPAR influences different cellular functions. In the present study, a highly metastatic colon cancer cell line, HCT116 was transfected with an expression vector containing a 5' uPAR cDNA fragment in an antisense orientation. This construct was most effective in reducing uPAR cell surface expression as confirmed by flow cytometry analysis. Antisense transfection of HCT116 cells had no effect on proliferation but the following effects were observed: (1) a 1.3-fold decreased adhesion; (2) a two fold decreased Erk MAP kinase activity; (3) a 2.7-fold decrease in Src kinase activity; (4) a 1.5- and two-fold decrease in uPA cell surface expression and secretion; (5) abrogation of promatrix metalloproteinase-9 secretion; and (6) a complete suppression of plasminogen-dependent matrix degradation. Using proteomic analysis, we demonstrate loss of approximately 200 proteins and quantitative differences in the expression of 141 other proteins in an antisense-clone compared to wild-type and mock-transfected control. Such changes in protein expression with the down-regulation of uPAR may be an important contributor in colon cancer progression and metastasis and may not only provide a basis to develop a proteomic data bank of uPAR-mediated signaling molecules but may also lead to the development of therapeutic approaches for the cure and better management of colon cancer. PMID- 12627385 TI - Mimicry of dimerization by synthetic peptides designed to target homologous regions of proteins. AB - Rapid progress in sequencing various genomes has highlighted the need for the development of biochemical reagents for the detection of thousands of expressed gene products. The magnitude of this detection problem exceeds current technical capabilities. In an attempt to address this shortcoming, a novel approach has been developed called mimicry of dimerization. Peptide tags have been designed to bind to a specific region of parvalbumin on the basis of amino acid sequence homology with this segment. Multivalent ligands were produced by coupling the synthetic peptides to activated dextran polymers and binding was assessed by chemiluminescence of enhanced avidity reactions using a high density of target protein at the binding surface. Binding of the peptide ligands to parvalbumin was strongest under assay conditions that enriched for native monomeric protein and was affected by pH, temperature and solvent conditions. The results suggest that it should be possible to develop specific reagents for tagging proteins on the basis of sequence and secondary structure information. PMID- 12627386 TI - Drug induced proteome changes in Candida albicans: comparison of the effect of beta(1,3) glucan synthase inhibitors and two triazoles, fluconazole and itraconazole. AB - The dimorphic fungus Candida albicans is an opportunistic human pathogen. Candidiasis is usually treated with azole antifungal agents. However clinical treatments may fail due to the appearance of resistance to this class of antifungal agents in Candida. Echinocandin derivatives are an alternative for the treatment of these fungal infections and are active against azole resistant isolates of C. albicans. Azoles inhibit the lanosterol 14 alpha demethylase which is a key enzyme in the synthesis of ergosterol. In contrast, the echinocandin class of antibiotics inhibit noncompetitively beta-(1,3)-D-glucan synthesis in vitro. We have investigated the impact of mulundocandin on the proteome of C. albicans and compared it to those of a mulundocandin derivative, as well as to two azoles of different structure, fluconazole and itraconazole. The changes in gene expression underlying the antifungal responses were analyzed by comparative 2-D PAGE. Dose dependant responses were kinetically studied on C. albicans grown at 25 degrees C (yeast form) in synthetic dextrose medium. This study shows that antifungals with a common mechanism of action lead to comparable effects at the proteome level and that a proteomics approach can be used to distinguish different antifungals, with the promise to become a useful tool to study drugs of unknown mechanism of action. PMID- 12627390 TI - Vaccines: back to the future. PMID- 12627387 TI - Proteomic analysis of Lactococcus lactis, a lactic acid bacterium. AB - Lactococcus lactis is a Gram-positive bacteria, which belongs to the group of lactic acid bacteria among which several genera play an essential role in the manufacture of food products. Cytosolic proteins of L. lactis IL1403 cultivated in M17 broth have been resolved by two-dimensional gel electrophoresis using two pH gradients (pH 4-7, 4.5-5.5). More than 230 spots were identified by peptide mass fingerprints, corresponding to 25% of the predicted acid proteome. The present study made it possible to describe at the proteome level a significant number of cellular pathways (glycolysis, fermentation, nucleotide metabolism, proteolysis, fatty acid and peptidoglycan synthesis) related to important physiological processes and technological properties. It also indicated that the fermentative metabolism, which characterizes L. lactis is associated with a high expression of glycolytic enzymes. Thirty-four proteins were matched to open reading frames for which there is no assigned function. The comparison at the proteome level of two strains of L. lactis showed an important protein polymorphism. The comparison of the proteomes of glucose- and lactose-grown cells revealed an unexpected link between the nature of the carbon source and the metabolism of pyrimidine nucleotides. PMID- 12627391 TI - Antigens and structure of the adenovirus. Reprinted from J. Mol. Biol. 1965; 13: 13-20. PMID- 12627392 TI - Natural and synthetic sialic acid-containing inhibitors of influenza virus receptor binding. AB - Influenza viruses attach to susceptible cells via multivalent interactions of their haemagglutinins with sialyloligosaccharide moieties of cellular glycoconjugates. Soluble macromolecules containing sialic acid from animal sera and mucosal fluids can act as decoy receptors and competitively inhibit virus mediated haemagglutination and infection. Although a role for these natural inhibitors in the innate anti-influenza immunity is still not clear, studies are in progress on the design of synthetic sialic acid-containing inhibitors of receptor binding which could be used as anti-influenza drugs. PMID- 12627393 TI - SWI/SNF chromatin remodelling complex and retroviral gene silencing. AB - Because of the unique infectious cycle of retroviruses which involves the integration of the retroviral genome into the host chromosome, many cellular chromosomal proteins are used by the virus to maintain its gene expression. At the same time, cellular mechanisms for the surveillance and exclusion of non-self expression by such intragenomic parasites operate as an important host defence system in the cellular nuclei. Retroviruses have strategies for escaping from host defence systems, such as by maintaining or reactivating viral expression in specific host cell types. Understanding such epigenetical regulation would be essential for progress in retroviral virology. In this review, we emphasise the importance of the chromatin remodelling factor SWI/SNF complex as one of the key players in epigenetic regulation of host and viral gene expression. An understanding of these mechanisms will surely lead to new ideas on the pathogenicity of this virus, on the latent infection observed in many other viruses, and further forward the design of unique retroviral vectors for long term transgene expression, providing strong tools for human gene therapy and regenerative medicine. PMID- 12627394 TI - Cloning of herpesviral genomes as bacterial artificial chromosomes. AB - Herpesviruses, which are important pathogens for both animals and humans, have large and complex genomes with a coding capacity for up to 225 open reading frames (ORFs). Due to the large genome size and the slow replication kinetics in vitro of some herpesviruses, mutagenesis of viral genes in the context of the viral genome by conventional recombination methods in cell culture has been difficult. Given that mutagenesis of viral genes is the basic strategy to investigate function, many of the herpesvirus ORFs could not be defined functionally. Recently, a completely new approach for the construction of herpesvirus mutants has been developed, based on cloning of the virus genome as a bacterial artificial chromosome (BAC) in E. coli. This technique allows the maintenance of viral genomes as a plasmid in E. coli and the reconstitution of viral progeny by transfection of the BAC plasmid into eukaryotic cells. Any genetic modification of the viral genome in E. coli using prokaryotic recombination proteins is possible, thereby allowing the generation of mutant viruses and facilitating the analysis of herpesvirus genomes cloned as infectious BACs. In this review, we describe the principle of cloning a viral genome as a BAC using murine gammaherpesvirus 68 (MHV-68), a mouse model for gammaherpesvirus infections, as an example. PMID- 12627395 TI - Structural evidence for common functions and ancestry of the reovirus and adenovirus attachment proteins. AB - The crystal structure of the reovirus attachment protein, sigma1, reveals a fibre like structure that is remarkably similar to that of the adenovirus attachment protein, fibre. Both proteins are trimers with head-and-tail morphology. They share unique domain structures and functional properties including defined regions of flexibility within the tail and an unusual symmetry mismatch with the pentameric viral capsid protein into which they are inserted. Moreover, the receptors for reoviruses and adenoviruses, junctional adhesion molecule 1 and coxsackievirus and adenovirus receptor, respectively, also share key structural and functional properties. Although reoviruses and adenoviruses belong to different virus families and have few properties in common, the observed similarities between sigma1 and fibre point to a conserved mechanism of attachment and an ancient evolutionary relationship. PMID- 12627396 TI - Translational regulator RpL10p/Grc5p interacts physically and functionally with Sed1p, a dynamic component of the yeast cell surface. AB - Biogenesis of an active ribosome complement and a dynamic cell surface complement are two major determinants of cellular growth. In yeast, the 60S ribosomal subunit protein RpL10p/Grc5p functions during successive stages in ribosome biogenesis, specifically rRNA processing, nucle(ol)ar preribosomal subunit assembly, nucleo-cytoplasmic transport and cytoplasmic maturation of ribosomes. Here, we report that a two-hybrid screen identified yeast genes SED1, ACS2 and PLB3 as encoding proteins physically interacting with both ribosomal RpL10p/Grc5p and its human homologue hRpL10p/QMp. SED1 encodes a differentially expressed cell wall protein which is proposed to be first transiently secreted to the plasma membrane as a GPI (glycosylated derivative of phosphoinositol)-anchored form and to be then transferred to the glucan layer of the cell wall. Ectopic expression of SED1 rescues both the aberrant growth phenotype and the translation defect of grc5-1(ts) temperature-sensitive cells. Furthermore, we report that Sed1p associates with translating ribosomes suggesting a novel, cytoplasmic role for Sed1p. ACS2 encodes one of the two yeast acetyl-CoA synthases and represents a key enzyme in one of several metabolic routes to produce acetyl-CoA, which in turn is indispensable for lipid biosynthesis. PLB3 encodes a phospholipase, which is active in the breakdown of membrane lipids. Our results support the view that Grc5p/RpL10p links ribosome function to membrane turnover and cell surface biogenesis. PMID- 12627397 TI - Proteome analysis of recombinant xylose-fermenting Saccharomyces cerevisiae. AB - Introduction of an active xylose utilization pathway into Saccharomyces cerevisiae, which does not naturally ferment pentose sugars, is likely to have a major impact on the overall cellular metabolism as the carbon introduced to the cells will now flow through the pentose phosphate pathway. The metabolic responses in the recombinant xylose-fermenting S. cerevisiae were studied at the proteome level by comparative two-dimensional gel electrophoresis of cellular proteins within a pH range of 3-10. Glucose-limited chemostat cultivations and corresponding chemostat cultivations performed in media containing xylose as the major carbon source were compared. The cultivations were studied in aerobic and anaerobic metabolic steady states and in addition at time points 5, 30 and 60 min after the switch-off of oxygen supply. We identified 22 proteins having a significant abundance difference on xylose compared to glucose, and 12 proteins that responded to change from aerobic to anaerobic conditions on both carbon sources. On xylose in all conditions studied, major changes were seen in the abundance of alcohol dehydrogenase 2 (Adh2p), acetaldehyde dehydrogenases 4 and 6 (Ald4p and Ald6p), and DL-glycerol 3-phosphatase (Gpp1p). Our results give indications of altered metabolic fluxes especially in the acetate and glycerol pathways in cells growing on xylose compared to glucose. PMID- 12627398 TI - Four ARF GAPs in Saccharomyces cerevisiae have both overlapping and distinct functions. AB - Previous studies in yeast have revealed the presence of four proteins with a conserved, cysteine-rich, ARF GAP domain that share the ability to suppress the conditional growth defect of the arf1-3 mutant. Three of these proteins have been shown previously to be ADP-ribosylation factor (ARF) GTPase-activating proteins (GAPs). We now demonstrate that the fourth also exhibits in vitro ARF GAP activity and correlates the suppressor and ARF GAP activities for all four. Because the four ARF GAP proteins are quite diverse outside the ARF GAP domain, a genetic analysis was undertaken to define the level of functional cross-talk between them. A large number of synthetic defects were observed that point to a high degree of functional overlap among the four ARF GAPs. However, several differences were also noted in the ability of each gene to suppress the synthetic defects of others and in the impact of single or combined deletions on assays of membrane traffic. We interpret these results as supportive evidence for roles of ARF GAPs in a number of distinct, essential cellular processes that include cell growth, protein secretion, endocytosis and cell cycling. The description of the specificities of the ARF GAPs for the different responses is viewed as a necessary first step in dissecting biologically relevant pathways through a functionally overlapping family of signalling proteins. PMID- 12627399 TI - Viable Saccharomyces cerevisiae cells at high concentrations cause early growth arrest of non-Saccharomyces yeasts in mixed cultures by a cell-cell contact mediated mechanism. AB - The growth of Kluyveromyces thermotolerans and Torulaspora delbrueckii was examined in mixed cultures with Saccharomyces cerevisiae in YPD modified for wine fermentations. Although the three yeasts had similar maximum specific growth rates in these fermentations, K. thermotolerans and T. delbrueckii arrested growth earlier than S. cerevisiae, thereby obtaining lower stationary phase cell concentrations than S. cerevisiae. Various single and mixed culture fermentations with the three yeasts were carried out in order to find an explanation for this phenomenon. The early growth arrests of K. thermotolerans and T. delbrueckii were absent in single cultures of the two yeasts, and they seemed to be due neither to nutrient limitations nor to the presence of growth-inhibitory compounds. Rather, they seemed to be due to a cell-cell contact mechanism dependent on the presence of viable S. cerevisiae cells at high concentrations. These results contribute to an increased understanding of why K. thermotolerans and T. delbrueckii arrest growth before S. cerevisiae during wine fermentations. PMID- 12627400 TI - RHO1 (YlRHO1) is a non-essential gene in Yarrowia lipolytica and complements rho1Delta lethality in Saccharomyces cerevisiae. AB - The synthesis of beta-1,3-glucan, the structural component of the yeast cell wall that gives shape to the cell, occurs at the plasma membrane and is the result of the activity of at least a two-component complex. Fks1p is the catalytic subunit directly responsible for the synthesis of beta-1,3-glucan, whilst the second subunit, Rho1p, has a GTP-dependent regulatory role (Yamochi et al., 1994). RHO1 has been characterized in Saccharomyces cerevisiae (Yamochi et al., 1994), and in several other fungal species. In this work, we have used degenerate oligonucleotides derived from the conserved regions of Rho1ps to isolate the RHO1 gene of Yarrowia lipolytica. The gene isolated in this way, which we have named YlRHO1, encodes a 204 amino acid protein that shows a high degree of homology with other Rho1ps. However, unlike S. cerevisiae, the ylrho1Delta disruptant strain in Y. lipolytica is viable, although it exhibits an increased sensitivity to Calcofluor white and Congo red. Also, YlRHO1 complements rho1 lethality in S. cerevisiae at both 28 degrees C and 37 degrees C. The complete sequence of YlRHO1 can be obtained from GenBank under Accession No. AF279915. PMID- 12627401 TI - Cyclic AMP mediates the cell cycle dynamics of energy metabolism in Saccharomyces cerevisiae. AB - We have investigated the role of 3',5'-cyclic-adenosine-monophosphate (cAMP) in mediating the coupling between energy metabolism and cell cycle progression in both synchronous cultures and oscillating continuous cultures of Saccharomyces cerevisiae. For the first time, a peak in intracellular cAMP was shown to precede the observed breakdown of trehalose and glycogen during cell cycle-related oscillations. Measurements in synchronous cultures demonstrated that this peak can be associated with the cell cycle dynamics of cAMP under conditions of glucose-limited growth, which was found to differ significantly from that observed in synchronous glucose-repressed cultures. Our results support the notion that cAMP plays a major role in mediating the integration of energy metabolism and cell cycle progression, both in the single cell and during cell cycle-related oscillations in continuous culture, respectively. Evidence is presented that the dynamic behaviour of intracellular cAMP during the cell cycle is modulated depending on nutrient supply. The implications of these findings regarding the role of cAMP in regulating cell cycle progression and energy metabolism are discussed. PMID- 12627402 TI - Integrating issues of efficacy, safety, and cost-effectiveness. Proceedings of the Fourth International Meeting on statistics Methods in Biopharmacy. Paris, France. September 24-25, 2001. PMID- 12627403 TI - Population pharmacokinetics/pharmacodynamics relationships of an anticancer drug. AB - This paper proposes a method for studying the toxicity of an anticancer drug with a delayed effect. The goal is to predict a dosage regimen with controlled toxicity. To this end, a semi-physiological model is used. A limit of toxicity is demonstrated, which is intrinsic to the model. It reduces the effect of high drug concentrations. This limit explains the mixed behaviour of the drug: time dependence and concentration-dependence, according to the dose actually administered. A population analysis is performed to estimate the parameters of the model, and to predict a safe dosage regimen. PMID- 12627404 TI - Identifying effective and/or safe doses by stepwise confidence intervals for ratios. AB - Typical randomized clinical dose-finding studies consist of the comparison of several doses of a drug versus a placebo. Interest lies in estimating relevant doses among those under investigation for efficacy and safety variables, such as the minimum effective dose or the maximum safe dose (or estimating both doses simultaneously). Step-down procedures have been proposed for comparing the standardized differences of the dose groups against placebo. In this paper we consider the ratio of population means and propose stepwise confidence intervals for these ratios. These confidence intervals do not require multiplicity adjustments and yield the same decisions as the associated test procedures. In addition, several power concepts are investigated within the present framework. The results allow sample size determination in the design phase of a study for the probability of estimating correctly the dose of interest. Auxiliary results of a numerical study show the range of application of these methods. PMID- 12627405 TI - Two-treatment comparison based on joint toxicity and efficacy ordered alternatives in cancer trials. AB - The primary goal of anticancer treatments is to attain efficacy, however toxicity could affect the course of the therapy. Methods have been proposed for comparing two treatments on the basis of the joint distribution for safety and efficacy outcomes, but they do not take into account the cumulative doses of drugs (chemotherapy) or radiation (radiotherapy) received by each patient. Moreover, these methods assume a parametric form for the joint distribution. In this paper we define a multi-dimensional parameter including toxicity and efficacy outcomes and the dose at which one, none or both occur. Each patient is classified into an ordered category depending on the order of occurrence of these two criteria: the sooner the patient benefits from efficacy and/or the later he/she experiences toxicity, the better is the treatment. We then apply likelihood ratio tests with ordered alternatives. This procedure requires constrained maximum likelihood estimation via isotonic regression. A large set of simulations compares the proposed tests to other more usual tests and the results show a good power and a satisfactory type I error control. Our approach is illustrated with a multi centre randomized clinical trial involving patients with metastatic non seminomatous germ cell tumours. PMID- 12627406 TI - Non-parametric estimators of a monotonic dose-response curve and bootstrap confidence intervals. AB - In this paper we consider study designs which include a placebo and an active control group as well as several dose groups of a new drug. A monotonically increasing dose-response function is assumed, and the objective is to estimate a dose with equivalent response to the active control group, including a confidence interval for this dose. We present different non-parametric methods to estimate the monotonic dose-response curve. These are derived from the isotonic regression estimator, a non-negative least squares estimator, and a bias adjusted non negative least squares estimator using linear interpolation. The different confidence intervals are based upon an approach described by Korn, and upon two different bootstrap approaches. One of these bootstrap approaches is standard, and the second ensures that resampling is done from empiric distributions which comply with the order restrictions imposed. In our simulations we did not find any differences between the two bootstrap methods, and both clearly outperform Korn's confidence intervals. The non-negative least squares estimator yields biased results for moderate sample sizes. The bias adjustment for this estimator works well, even for small and moderate sample sizes, and surprisingly outperforms the isotonic regression method in certain situations. PMID- 12627407 TI - Assessing non-inferiority of a new treatment in a three-arm clinical trial including a placebo. AB - In non-inferiority trials, where non-inferiority of a new experimental drug compared to an active control has to be shown, it may be advisable to use an additional placebo group for internal validation if ethically justifiable. The focus of this paper is on such designs. Assuming normality and homogeneity of variances we will derive a statistical test procedure which turns out to be equivalent to the assessment based on Fieller's confidence interval. Based on the power function of this test, sample size calculations are carried out to achieve a given power. Additionally, the optimal allocation of the total sample size is derived. As an alternative to this parametric procedure, the bootstrap percentile interval is discussed and finally compared with Fieller's confidence interval in a study on mildly asthmatic patients. PMID- 12627408 TI - Assessing individual bioequivalence using the structural equation model. AB - The structural equation model (SEM) is introduced as a useful approach for assessing individual bio-equivalence. SEM parameters are estimated using a partial likelihood analysis and the hypotheses of individual bioequivalence is evaluated in a disaggregate way, testing separately the hypothesis concerning SEM parameters, and assessing the overall hypothesis of individual bioequivalence using the intersection-union principle. Limits of bioequivalence for SEM parameters are proposed and a power analysis is carried out. PMID- 12627409 TI - Kullback-Leibler divergence for evaluating bioequivalence. AB - In this paper we propose a methodology for evaluating the bioequivalence of two formulations of a drug that encompasses not only average bioequivalence (ABE), but also the more recently introduced measures of population bioequivalence (PBE) and individual bioequivalence (IBE). The latter two measures are concerned with prescribability (PBE) and switchability (IBE). The main idea is to use the Kullback-Leibler divergence (KLD) as a measure of discrepancy between the distributions of the two formulations. Two formulations are declared bioequivalent if the upper bound of a level-alpha confidence interval for the KLD is less than a given goalpost to be set by a regulator. This new methodology overcomes many of the disadvantages of the corresponding measures recommended by the FDA. In particular the KLD: (i) possesses the natural hierarchical property that IBE => PBE => ABE; (ii) satisfies the properties of a true distance metric; (iii) is invariant to monotonic transformations of the data; (iv) generalizes easily to the multivariate case where equivalence on more than one parameter (for example, AUC, C(max) and T(max)) is required; and (v) is applicable over a wide range of distributions of the response variable (for example, those in the exponential family). The performance of the KLD relative to the metric proposed in guidance by the FDA for the evaluation of individual bioequivalence is evaluated using a simulation study. Previously published retrospective analyses using the FDA-proposed metric are contrasted with those based on the KLD. It is concluded that the KLD is a viable alternative to the FDA-proposed metric and that its mathematical and statistical properties make it a readily interpretable measure of the differences between formulations. PMID- 12627410 TI - A score test for establishing non-inferiority with respect to short-term survival in two-sample comparisons with identical proportions of long-term survivors. AB - In recent years randomized trials designed to establish non-inferiority of a new treatment as compared to a standard one have been more widely used. Two-sample statistics have been proposed for this equivalence testing problem. However, they are not suited to situations where a long-term survivor fraction is expected. In this paper we propose a score test designed for establishing non-inferiority for the new treatment as compared to the standard one while assuming identical long term survivor rates. Simulations results show that the proposed statistic has satisfactory size and power as long as certain restricting conditions are verified. A breast cancer trial is analysed as an example. PMID- 12627412 TI - Issues in designing flexible trials. AB - We outline the general framework of adaptive combination tests and discuss their relationship to flexible group sequential designs. An important field of applications is sample size reassessment. We discuss reassessment rules based on conditional power arguments using either the observed or the prefixed effect size. These rules tend to lead to large expected sample sizes for small actual effects. However, the application of a maximal bound for the second stage sample size leads to more favourable properties. Additionally, we consider an optimized reassessment rule in terms of expected sample sizes. Since the adaptive design does not use the classical test statistics for some types of sample size reassessments, the adaptive test may reject the null hypothesis while the classical one-sample test does not. We characterize sample size reassessment rules, where such inconsistencies are avoided. Finally, the extension of flexibility to the number of stages is explored. In the first interim analysis a second interim analysis is only planned if the chance to achieve a decision there is high. This leads to savings in the average number of interim analysis performed, without paying a noticeable price in terms of expected sample size. PMID- 12627411 TI - The use of putative placebo in active control trials: two applications in a regulatory setting. AB - For life-threatening diseases, ethical considerations preclude the inclusion of an untreated control group in the investigation of a new therapeutic agent when a standard therapy exists. In these cases, active controlled studies are conducted, and may be planned to demonstrate either superiority or equivalence/non inferiority of the new drug over the standard therapy (active control). In the non-inferiority study, an important aspect is the ability to detect an inferior drug (assay sensitivity). It has been suggested that assay sensitivity for a non inferiority study should be deduced from historical data, specifically placebo controlled studies with the standard therapy. The assessment of assay sensitivity may also be important in a superiority trial that fails to demonstrate a statistically significant difference between treatments, and the sponsor attempts to determine whether there is lack of inferiority as an alternative hypothesis for regulatory approval. This paper describes two methods of putative placebo analysis for assessing assay sensitivity in active controlled trials. One approach imputes a point estimate for the odds ratio (95 per cent confidence interval) for a new drug (T) compared to a placebo control (P). A Bayesian approach calculates the posterior probability that T is superior to P, or, that T is at least k per cent as good as the active control (A) and A is more effective than P. These methods are applied in two clinical/regulatory settings: a phase III trial comparing docetaxel (Taxotere) to doxorubicin in metastatic breast cancer patients, and a phase III programme with two trials comparing enoxaparin (Lovenox) plus aspirin to unfractionated heparin plus aspirin in patients with unstable angina or non-Q-wave myocardial infarction. The methodologies presented in this paper were used in securing regulatory approval for docetaxel in the treatment of locally advanced or metastatic breast cancer after failure of prior chemotherapy, and for enoxaparin in the treatment of acute coronary syndrome. PMID- 12627413 TI - Mid-course sample size modification in clinical trials based on the observed treatment effect. AB - It is not uncommon to set the sample size in a clinical trial to attain specified power at a value for the treatment effect deemed likely by the experimenters, even though a smaller treatment effect would still be clinically important. Recent papers have addressed the situation where such a study produces only weak evidence of a positive treatment effect at an interim stage and the organizers wish to modify the design in order to increase the power to detect a smaller treatment effect than originally expected. Raising the power at a small treatment effect usually leads to considerably higher power than was first specified at the original alternative. Several authors have proposed methods which are not based on sufficient statistics of the data after the adaptive redesign of the trial. We discuss these proposals and show in an example how the same objectives can be met while maintaining the sufficiency principle, as long as the eventuality that the treatment effect may be small is considered at the design stage. The group sequential designs we suggest are quite standard in many ways but unusual in that they place emphasis on reducing the expected sample size at a parameter value under which extremely high power is to be achieved. Comparisons of power and expected sample size show that our proposed methods can out-perform L. Fisher's 'variance spending' procedure. Although the flexibility to redesign an experiment in mid-course may be appealing, the cost in terms of the number of observations needed to correct an initial design may be substantial. PMID- 12627415 TI - An illustration of the modelling of cost and efficacy data from a clinical trial. AB - Health care providers, purchasers and policy makers need to make informed decisions regarding the provision of cost-effective care. When a new health care intervention is to be compared with the current standard, an economic evaluation alongside an evaluation of health benefits provides useful information for the decision making process. We consider the information on cost-effectiveness which arises from an individual clinical trial comparing the two interventions. Recent methods for conducting a cost-effectiveness analysis for a clinical trial have focused on the net benefit parameter. The net benefit parameter, a function of costs and health benefits, is positive if the new intervention is cost-effective compared with the standard. In this paper we describe frequentist and Bayesian approaches to cost-effectiveness analysis which have been suggested in the literature and apply them to data from a clinical trial comparing laparoscopic surgery with open mesh surgery for the repair of inguinal hernias. We extend the Bayesian model to allow the total cost to be divided into a number of different components. The advantages and disadvantages of the different approaches are discussed. In January 2001, NICE issued guidance on the type of surgery to be used for inguinal hernia repair. We discuss our example in the light of this information. PMID- 12627414 TI - Blinded sample size reassessment in non-inferiority and equivalence trials. AB - Even in situations where the design and conduct of clinical trials is highly standardized, there may be a considerable between-study variation in the observed variability of the primary outcome variable. As a consequence, performing a study in a fixed sample size design implies a considerable risk of resulting in a too high or too low sample size. This difficulty can be alleviated by applying a design with internal pilot study. After a provisional sample size calculation in the planning stage, a portion of the planned sample is recruited and the sample size is recalculated on the basis of the observed variability. To comply with the requirement of some regulatory guidelines only blinded data should be used for the reassessment procedure. Furthermore, the effect on the type I error rate should be quantified. The current literature presents analytical results on the actual level in the t-test situation only for superiority trials. In these situations, blinded sample size recalculation does not lead to an inflation of the type I error rate. We extended the methodology to non-inferiority and equivalence trials with normally distributed outcome variable and hypotheses formulated in terms of the ratio and difference of means. Surprisingly, in contrast to the case of testing superiority, we observed actual type I error rates above the nominal level. The extent of inflation depends on the required sample size, the sample size of the internal pilot study, and the standardized equivalence or non-inferiority margin. It turned out that the elevation of the significance level is negligible for most practical situations. Nevertheless, the consequences of sample size reassessment have to be discussed case by case and regulatory concerns with respect to the actual size of the procedure cannot generally be refuted by referring to the fact that only blinded data were used. PMID- 12627416 TI - A Bayesian analysis of pharmacoeconomic data from a clinical trial on schizophrenia. AB - Pharmacoeconomic studies are performed in a higher frequency to assess the economic interest of new drugs. However, a standard methodology does not still completely exist. We present here the principles and results of a cost effectiveness Bayesian analysis on data from 146 patients (interim analysis) collected during a clinical trial. This trial was originally planned to enrol 245 patients with predominantly negative schizophrenia symptoms and involved four treatment groups (a new treatment given at low dose and high dose, a comparator and a placebo). First, some prior distributions of the cost-effectiveness ratio were numerically deduced from the effectiveness parameter clinical priors (based on investigators' opinions and questionnaires before going to blind breaking) and from cost function priors. The costs taken into account were hospitalizations, sick leave days, treatments, visits to the doctor, laboratory exams and suicide attempts. The effectiveness parameter was the change from baseline on SANS (scale for the assessment of negative symptoms). Posterior distributions were elaborated for the cost-effectiveness ratio by combining the cost-effectiveness ratio priors and likelihood together using the Bayes theorem. Results lead to a conclusion in favour of the new treatment given at high dose. PMID- 12627417 TI - Antenatal urodynamic studies in the fetal lamb: experimental protocol and preliminary results. AB - OBJECTIVES: To set up a fetal lamb model for intrauterine fetal urodynamic studies. METHODS: Fourteen fetal lambs underwent placement of a bladder catheter at a mean gestational age of 87 days. Three fetuses also had a partial urethral obstruction by the simultaneous placement of a peri-urethral constricting ring. Urodynamic and ultrasound studies were performed weekly by the filling cystometry method. RESULTS: Hundred and six voiding cycles were recorded during 25 urodynamic studies between 84- and 133-days gestation. All voiding profiles were biphasic with a mean duration of 4.2 min (range 1-10), a mean voiding pressure of 23 cm of water (range 7-33) and a mean periodicity of 19.2 min (range 11-50). The obstructed animals had bladder overactivity. This correlated with ultrasound and post-mortem findings of megacystis and bilateral hydroureteronephrosis. The fetal mortality rate was 85.7% and the mean duration of survival post surgery was 45 +/ 5.7 days. CONCLUSION: Serial urodynamic studies could be performed in a fetal lamb model. Following partial urethral obstruction, bladder overactivity was observed. PMID- 12627418 TI - Isolated pericardial effusion in the human fetus: a report of three cases. AB - OBJECTIVE: Our objective was to determine the possible underlying etiologies and outcome in isolated fetal pericardial effusion. METHODS: Doppler fetal echocardiography allowed the diagnosis of pericardial effusion in three patients and revealed the etiology in two. RESULTS: We present the findings in three cases of isolated pericardial effusion. In the first, the pericardial effusion was a manifestation of trisomy 21 associated with a myeloproliferative disorder. In the second, the pericardial fluid collection was the first sign of an autosomal recessive disease, idiopathic infantile arterial calcification. The third case was remarkable because of the spontaneous resolution of a large pericardial fluid collection. CONCLUSION: Isolated fetal pericardial effusion covers a wide spectrum of etiologies from severe genetic and chromosomal diseases to transient forms. PMID- 12627419 TI - Congenital limb deficiencies in newborn infants: prevalence, characteristics and prenatal diagnosis. AB - Congenital limb deficiency (CLD) occurs in 0.54 to 0.59/1000 live-born infants and varies according to its anatomic location, type and cause. Our aim was to present a fetus with a prenatal ultrasonographic diagnosis of CLD (transverse reduction deficiency of the left upper limb), and to determine the prevalence and clinical characteristics of CLD at a tertiary medical centre in Israel. Among 78 500 live-born infants, there were 24 cases of CLD (0.31/1000): 45.8% of the CLD cases with affected upper limbs, 45.8% with affected lower limbs, and 8.4% with both limbs affected; 88.4% of the limb deficiencies were longitudinal and 11.6% were transverse; and, 33.3% (8/24) of the affected newborn infants had additional congenital anomalies. We conclude that CLD is not an infrequent finding in live born infants. Comprehensive ultrasonography of the fetus allows early prenatal diagnosis of CLD and provides the parents with important information and helps them in their decision regarding the fate of the pregnancy. PMID- 12627420 TI - Development and targeted application of a rapid QF-PCR test for sex chromosome imbalance. AB - OBJECTIVES: A QF-PCR test has been developed to diagnose sex chromosome imbalances in prenatal samples and has been applied to a diagnostic service. METHODS: The test uses a PCR multiplex with eight primer pairs: six X-chromosome polymorphic markers, including two markers from Xp (a region not included in previously published sex chromosome aneuploidy tests), one polymorphic marker for a locus common to the long arms of the X and Y chromosomes, and the non polymorphic amelogenin marker. Homozygosity for all X-chromosome markers and the absence of the Y-chromosome amelogenin marker is highly likely (907 : 1) to represent monosomy X (Turner syndrome), but interphase FISH is always used to confirm such a result. RESULTS: Blind studies were carried out to validate the test and the first year of clinical use has been reported. Results are usually issued within one working day, and the test is more efficient than interphase FISH. CONCLUSIONS: The sex chromosome imbalance test has been targeted to prenatal samples displaying a clear ultrasound indication consistent with Turner syndrome, and has also been used to identify fetal sex in pregnancies at risk of inheriting a sex-linked molecular disorder. No misdiagnoses were made. It is concluded that QF-PCR can rapidly and accurately diagnose sex chromosome status and imbalances, reducing maternal anxiety and aiding in efficient pregnancy management. PMID- 12627421 TI - Mucolipidosis III type C: first-trimester biochemical and molecular prenatal diagnosis. AB - OBJECTIVES: Mucolipidosis IIIC (MLIIIC) is a rare autosomal recessive lysosomal storage disease resulting from defective mannose 6-phosphate-dependent lysosomal enzyme trafficking; mutations of the gamma subunit of N-acetylglucosamine-1 phosphotransferase (GINAcPT) were recently found to cause its pathogenesis. We report here for the first time prenatal diagnosis (PND) for MLIIIC by means of chorionic villous sampling (CVS). METHODS AND RESULTS: A fetus in a large Bedouin Moslem family was found to be homozygous for the founder haplotype and the mutational SSCP pattern of MLIIIC. The diagnosis was confirmed by markedly reduced lysosomal enzyme activities in cultured chorionic villi. The molecular identification of the disease-causing mutation in this large Bedouin-Moslem kindred permitted, for the first time, identification of carriers and couples at risk. CONCLUSIONS: The feasibility of early PND for a progressive disabling disease is important for its prevention. Nevertheless, the feasibility of PND raises a serious dilemma since affected individuals might have a variable phenotype and the disease is progressive and non-lethal. In addition, religious and social constraints are important factors to be taken into consideration in the genetic counseling of couples at risk. PMID- 12627422 TI - Prenatal diagnosis of a karyotypically normal pregnancy in a mother with a supernumerary neocentric 13q21 -->13q22 chromosome and balancing reciprocal deletion. AB - An adult female patient with a history of miscarriages was found to be carrying a stable supernumerary chromosome. The patient also carried a reciprocal paracentric deletion in chromosome 13q21/22. Microdissection and reverse fluorescence in situ hybridization FISH revealed that this supernumerary chromosome was derived from region 13q21 --> 13q22. The presence of a neocentromere on this supernumerary chromosome was confirmed by the absence of detectable alpha satellite DNA using FISH and the presence of centromere proteins CENP-C and CENP-A using immunofluorescence. The absence of telomere sequences suggests that the marker is a ring chromosome (r(13)). FISH using ordered BACs from the chromosome region 13q21 --> 13q31 permitted the precise positioning of the r(13) chromosome and the corresponding deletion to chromosome bands 13q21.32 -> 13q22.2. BAC 280J7 from within the r(13) was used as a FISH probe for the prenatal analysis of amniocytes at 16 weeks of gestation, which revealed a normal karyotype for the fetus. This r(13) chromosome represents the first description of chromosome 13 of the rarer class of neocentric chromosomes that are derived from interstitial deletions. It represents the first example of prenatal diagnosis in a phenotypically normal female that was ascertained to carry a neocentric marker. The presence of such a neocentric marker/deletion karyotype in a parent presents unique possible karyotypic outcomes for conceptions and unusual challenges for genetic counseling. PMID- 12627423 TI - Successful early in utero management of fetal hydrothorax in a twin pregnancy. AB - We present a case of dichorionic diamniotic twin pregnancy in which one of the fetuses was found to have a major pleural effusion at 15 weeks of gestation. A single-needle pleural fluid aspiration was performed at 15 and 16 weeks, but the fluid reaccumulated quickly after each procedure and at 16 weeks, the fetus was found to become progressively hydropic. A shunt was then successfully inserted at 17 weeks, which is the earliest gestation reported so far in the literature for such a procedure to treat isolated hydrothorax. Because we felt that the fetus would be too small for a classical double-pigtail pleuroamniotic shunt, we used a multilength double-pigtail bladder stent (Harrison drain; Cook; Spencer; Indiana; USA) via a 13-gauge echo tip trocar. This shunt could be used for both singleton and twin pregnancies presenting with fetal pleural effusion from as early as 16 to 17 weeks to prevent the development of fetal hydrops and polyhydramnios and subsequent premature delivery. Treatment at this stage of gestation would also minimize the risk of lung hypoplasia, which is the main clinical issue when shunts are inserted after 24 weeks. PMID- 12627424 TI - Assessment of cortical gyrus and sulcus formation using MR images in normal fetuses. AB - OBJECTIVES: The purpose of this study was to estimate the development of the gyrus and sulcus formation in normal fetuses on the basis of the neuroanatomical findings using MR images in relation to gestational age. METHODS: The MR images were obtained from 109 normal fetuses from 18 to 39 weeks of gestation with no neurological problems. The MR images were classified into 8 stages of development for the gyrus and sulcus formation in the frontal and temporal lobes on the basis of the neuroanatomical findings reported by Chi et al. (1977) and Dorovini-Zis and Dolman (1977). We examined retrospectively the relationship between our classification and gestational age in comparison with the five-stage classification proposed by McArdle et al. (1987). RESULTS: There were significant differences in the gestational age among the 8 groups (P < 0.001). Multiple comparison of individual groups revealed significant differences in the gestational age among the groups (P < 0.05). Images from 28 to 34 weeks of gestation were classified into 4 stages in our classification, while being covered by one stage in McArdle's classification. CONCLUSION: Our classification is useful for the assessment of fetal cerebral maturation during the third trimester of pregnancy and may contribute to the prenatal diagnosis of developmental delay of the gyrus and sulcus formation. PMID- 12627425 TI - Huntington disease-unaffected fetus diagnosed from maternal plasma using QF-PCR. AB - The discovery of fetal DNA in maternal plasma from early pregnancies has led to new opportunities for clinical application. In the last few years there have been numerous reported applications, mainly fetal gender and RhD genotyping. The prenatal diagnosis of some inherited genetic diseases such as Huntington disease is also very frequently required in the prenatal diagnosis routine. We have successfully diagnosed, with a non-invasive procedure, an unaffected HD fetus at the 13th week of gestation using fetal DNA from maternal plasma and the quantitative fluorescent PCR method, which is one of the most sensitive ways to detect fetal DNA in maternal plasma at such an early time of gestation. PMID- 12627426 TI - Replicate real-time PCR testing of DNA in maternal plasma increases the sensitivity of non-invasive fetal sex determination. AB - BACKGROUND: We determined fetal sex in pregnancies referred for invasive prenatal diagnosis procedures by analysis of DNA in maternal plasma. METHODS: Twelve pregnancies at risk of X-linked haemophilia and 32 pregnancies at risk of chromosomal aneuploidies at a gestational age ranging from 10 to 18 weeks recruited before chorionic villus sampling or amniocentesis were involved in the study. Male fetal DNA in maternal plasma was detected by using real-time polymerase chain reaction with the SRY gene as a marker. RESULTS: The specificity of the system reached 100% (no Y signal was detected in 17 women pregnant with a female fetus) and the sensitivity reached 100% (SRY amplification in 27 examined samples). CONCLUSIONS: Amplification of free fetal DNA in maternal plasma is a valid and rapid technique for predicting fetal sex in first- and second-trimester pregnancies and could allow the restriction of invasive sampling procedures to male fetuses at risk of X-linked disorders. PMID- 12627427 TI - Mild fetal cerebral ventriculomegaly as a prenatal sonographic marker for Kartagener syndrome. AB - Primary ciliary dyskinesia (PCD), also referred to as immotile-cilia syndrome or Kartagener syndrome, is a group of genetic disorders caused by defective cilia leading to chronic sinupulmonary infection, situs inversus and reduced fertility. Some PCD patients also have cerebral ventriculomegaly or hydrocephalus. We report here two fetuses and one newborn with mild cerebral ventriculomegaly and a suspected and/or confirmed diagnosis of PCD. These cases demonstrate that mild fetal cerebral ventriculomegaly can be a prenatal sonographic marker of PCD, certainly in fetuses with situs inversus or a history of a previous sib with PCD. PMID- 12627428 TI - Maternal serum-integrated screening for trisomy 18 using both first- and second trimester markers. AB - OBJECTIVES: To estimate the prenatal screening performance of an integrated serum test for detecting trisomy 18, which combines measurements of first- and second trimester markers with maternal age to assign patient-specific risks. METHODS: Published and new observations of maternal serum marker levels in trisomy 18 and unaffected pregnancies are used to derive population parameters. These parameters are then combined in a multivariate Gaussian model to assign patient-specific risks for trisomy 18. RESULTS: The best combination of serum markers includes pregnancy-associated plasma protein-A in the first trimester and alpha fetoprotein, unconjugated estriol and human chorionic gonadotropin in the second trimester. At a second-trimester risk cutoff of 1 : 100, these 4 markers, in combination with maternal age, detect 90% of trisomy 18 pregnancies at a false positive rate of 0.1%. The odds of a trisomy 18 pregnancy among screen-positive women are 1 : 4. Without the first-trimester marker, detection is reduced to 67% at about the same false-positive rate. CONCLUSION: The algorithm described here is highly efficient for detecting trisomy 18 and should be considered by programs that offer serum-integrated screening for Down syndrome. PMID- 12627429 TI - First-trimester nuchal translucency measurement and echocardiography at 16 to 18 weeks of gestation in prenatal detection for trisomy 18. AB - BACKGROUND: Trisomy 18, the second most common autosomal trisomy, has the highest incidence of congenital heart disease of all chromosomal abnormalities. This study assessed the use of nuchal translucency (NT) measurement and fetal echocardiography at 16 to 18 weeks of gestation in prenatal detection for trisomy 18. METHODS: Screening for chromosomal aneuploidy using fetal NT measurement was performed at 10 to 14 weeks of gestation. Detailed fetal echocardiography was performed at 16 to 18 weeks of gestation immediately before genetic amniocentesis for fetal karyotyping in singleton pregnancies with increased fetal NT thickness. RESULTS: Of the 3151 singleton pregnancies included in our study, 171 cases (5.4%) of increased (> or =3.0 mm) NT were noted. Fetal chromosomal abnormalities were identified in 22 (12.9%) of these pregnancies, including 9 with trisomy 21, 5 with trisomy 18, 4 with 45,X and 4 with unbalanced structural abnormalities. Major defects of the heart and the great arteries were identified in 13 (7.6%) of these pregnancies with increased NT. These included eight pregnancies that also had the diagnosis of chromosomal aneuploidy. Among the 22 fetuses with confirmed aneuploidy, all 5 fetuses with trisomy 18, 1 of the 4 fetuses with 45,X and 2 of the 9 fetuses with trisomy 21 had increased fetal NT thickness associated with abnormal fetal echocardiography findings. CONCLUSIONS: Screening for Down syndrome and cardiac defects using first-trimester fetal NT measurement in combination with fetal echocardiography at 16 to 18 weeks of gestation is a feasible and sensitive procedure for the prenatal detection of trisomy 18. PMID- 12627430 TI - Comparison of models of maternal age-specific risk for Down syndrome live births. AB - OBJECTIVES: To display and compare the different published formulae that specify the association between maternal age and the risk of a Down syndrome live birth. METHODS: Papers published since 1987 on the prevalence of Down syndrome live births in relation to maternal age were located using MEDLINE and the references given in other papers. The data series and the models fitted to them were plotted to obtain a visual idea of their similarities and differences. RESULTS: The observed and modelled age-specific rates for Down syndrome births were remarkably similar in all published series of data for women up to the age of 35, were reasonably similar for women aged 35 to 45, but differed for women older than 45. CONCLUSION: In practice, the overall small differences in age-related risk between the different studies did not materially affect the performance of antenatal screening for Down syndrome. If a choice is to be made, the analysis based on the National Down Syndrome Cytogenetic Register (NDSCR) has marginal advantages since it is based on the largest data set and the corresponding model fits the data well. More data is needed to clarify the pattern of risk with maternal age among women over 45 years of age. PMID- 12627431 TI - Idiopathic fetal intrauterine growth restriction: a possible inheritance pattern. AB - OBJECTIVE: This study was conducted to assess if the delivery of a previous growth-retarded (IUGR) fetus increases the risk of having an IUGR fetus in subsequent pregnancies and to explore if a familial pattern of transmission is involved. METHODS: Seventy consecutive multiparous women whose fetus was IUGR (group 1) and 70 controls (group 2) were enrolled in this study. RESULTS: The proportion of women who developed preeclampsia (9 versus 2, p = 0.05) and who had delivered an IUGR fetus in a previous pregnancy (20 versus 4, p < 0.05) were higher in group 1 than in group 2. There was no difference in the incidence of chronic hypertension, diabetes, smoking, substance or alcohol abuse, and HIV infection between the groups. After adjustment for preeclampsia, the delivery of a previous IUGR fetus remained a risk factor for having a subsequent IUGR fetus [Odds ratio = 6.7 (CI 2.15-21.22), p < 0.01]. Pedigree analysis conducted in 15 families revealed a familial cluster of IUGR infants in all families that were investigated. In 9 out of 15 families, a dominant pattern of inheritance of IUGR was observed while the remaining families were more heterogeneous. In one family, a balanced carrier of chromosome 7 inversion generated a malformed fetus and two IUGR infants. CONCLUSIONS: This study clarifies that IUGR may be an inherited genetic condition and emphasizes that a knowledge of the family history and of the parental karyotype may be helpful in the prevention of both fetal malformations and adverse neonatal morbidity in subsequent low birth weight infants. PMID- 12627432 TI - Reversed diastolic flow in the middle cerebral artery: is it a terminal sign in a growth-retarded fetus? PMID- 12627436 TI - Peptide separations and dissociation constants in nonaqueous capillary electrophoresis: comparison of methanol and aqueous buffers. AB - Nonaqueous capillary electrophoresis was evaluated for its potential to separate peptides in methanolic background electrolytes in comparison to aqueous-methanol (50% v/v) and water. Isomeric aspartyl dipeptides and Leu- and Met-enkephalin served as model compounds. pK(a) values were determined in the three solvent systems based on the apparent pH scale and in the case of methanol additionally based on the conventional pH scale. Changing from water to methanol led to an increase of the ionization constants describing the dissociation equilibria of the carboxyl group and the amino group, respectively. The pK(a) shift was more pronounced for the carboxylic acid function leading to a compression of the mobility-pH curve. As reported for aqueous buffers, efficient separations of the peptides were achieved in methanolic background electrolytes including the resolution of the diastereomers of the isomeric alpha- and beta-aspartyl dipeptides. In contrast to aqueous buffers, the separation of Leu- and Met enkephalin could also be obtained in buffers in methanol at high pH. PMID- 12627437 TI - Physicochemical characterization of phosphinic pseudopeptides by capillary zone electrophoresis in highly acidic background electrolytes. AB - Phosphinic pseudopeptides (i.e., peptide isosteres with one peptide bond replaced by a phosphinic acid moiety) were analyzed and physicochemically characterized by capillary zone electrophoresis in the pH range of 1.1-3.2, employing phosphoric, phosphinic, oxalic and dichloroacetic acids as background electrolyte (BGE) constituents. The acid dissociation constant (pK(a)) of phosphinate group in phosphinic pseudopeptides and ionic mobilities of these analytes were determined from the pH dependence of their effective electrophoretic mobilities corrected to standard temperature and constant ionic strength of the BGEs. It was shown that these corrections are necessary whenever precise mobility data at very low pH are to be determined. Additionally, it was found that the ionic mobilities of the phosphinic pseudopeptides and pK(a) of their phosphinate group are affected by the BGE constituent used. The variability of migration behavior of the pseudopeptides can be attributed to their ion-pairing formation with the BGE components. PMID- 12627438 TI - Electrophoretic mobility for peptides with post-translational modifications in capillary electrophoresis. AB - Several semiempirical models for peptide electrophoretic mobility have been tested for capillary electrophoresis (CE) separation with a positively charged capillary using on-line CE combined with electrospray ionization-mass spectrometry (ESI-MS). In the current system with pH 2.7, the expression q/M(0.56) provided the best correlation with the electrophoretic mobility in the analysis of a set of 18 standard samples, where q is the calculated net charge and M is the molecular weight. The peptides resulting from various digests of horse heart myoglobin or bovine hemoglobin were used to demonstrate the validity of this correlation. Post-translationally modified peptides from tryptic digest of human myelin basic protein were also investigated and were found to provide excellent correlation with the linear plot when the total charge of the peptide was correctly calculated. If the total charge was not properly calculated then the post-translationally modified peptides fell off the linear plot. Using this method five arginine residues (residues 5, 49, 54, 97 and 130) were found to be partially citrullinated, four glutamine residues (residues 8, 103, 121 and 147) were found to be partially deamidated and both methionines at residues 21 and 167 were found to be partially oxidized. Three peptides were found with phosphorylation; TPPPSQGK (residue 98 to 105), FSWGAEGQR (residue 114 to 122), and SGSPMAR (residue 163 to 169) and Arginine at 107 was found to be partially monomethylated or dimethylated. The method may provide an excellent means of identifying the presence of peptides with post-translational modifications in conjunction with MS. PMID- 12627439 TI - Monitoring folding transitions of synthetic, branched-chain polypeptides by capillary zone electrophoresis. AB - The coil/helix transition of a synthetic, branched-chain polymeric polypeptide (poly (Lys(Glu(1)-DL-Ala(3))EAK), 50-Lys residues long in the backbone, as a function of increasing molarities of methanol in solution, is here studied by both, circular dichroism (CD) and capillary zone electrophoresis. CD spectra showed that, at 75% v/v methanol, the transition from random coil to fully helical structure was obtained, in a pH 1.1 HCI solution in the presence of 20 mM NaCI. CZE studies, run in parallel, exhibited the classical unfolding to folding sigmoidal transition, with mid-point at 60% v/v methanol concentration, plateauing at ca. 80% v/v organic solvent. Surprisingly, though, such unfolding to folding transition was accompanied by an expansion, rather than a contraction, of the resulting ordered polypeptide. As the charge of the polypeptide (a pure polycation at a pH of 2.1 in CZE) was kept rigorously constant, a plot of the radius of the polymer along the sigmoidal transition clearly showed that the radius of gyration of the helical, structured polypeptide was in fact larger than that of the random coil. Such results were confirmed by molecular dynamics simulations, which indicated that the dimensions of such polypeptide, in alpha helix configuration, were 8.5 nm (in length) and 3.2 nm (in diameter), whereas those of the corresponding random coil were 7.2 nm (in length) and 5.1 nm (length of shorter axis). It would thus appear that the randomized structure assumes the shape of a more compact object, roughly resembling a "rugby ball". PMID- 12627440 TI - Affinity capillary electrophoresis study of the linkage existing between proton and zinc ion binding to bacitracin A1. AB - Measurements by capillary electrophoresis (CE) of bacitracin A(1) effective mobility at different pH values permitted to estimate the five acidic dissociation constants and the Stokes radii at different protonation stages of the macrocyclic dodecapeptide. The pK(a) values were 3.6 and 4.4 for the two carboxylic groups of the lateral chains of D-Asp-11 and D-Glu-4, respectively, 6.4 for the aza-atom of the imidazole ring of His-10, 7.6 for the amino group of N-terminal Ile-1 and 9.7 for the delta-amino group of D-Orn-7, very close to the values obtained by other researchers by titration experiments. In agreement with a rigid macrocyclic structure the Stokes radii of different protonated forms ranged only between 14.3 and 14.8 A. Best fitting procedures performed on experimental mobility measured at two different pH values (5.50 and 6.72) in the presence of increasing Zn(+2) concentration allowed confirming the model that assumes the binding of Zn(+2) to P(0) peptide form with a 1.5 x 10(3) M(-1) intrinsic association constant. Following to Zn(+2) binding, the pK(a) of the amino group of N-terminal Ile-1 is shifted from 7.6 to 5.9 and the Stokes radius is reduced of about 3 A. The mean charge of the bacitracin A(1)-Zn(+2) complex resulted +1.67 and +1.12 at pH 5.50 and 6.72, respectively. These results suggest that the amino group of N-terminal Ile-1 is not essential for Zn(+2) binding. PMID- 12627441 TI - On-column derivatization of the antibiotics teicoplanin and ristocetin coupled to affinity capillary electrophoresis. AB - Binding constants between the glycopeptides teicoplanin (Teic) and ristocetin (Rist) and their derivatives to D-Ala-D-Ala terminus peptides were determined by on-column receptor synthesis coupled to partial-filling affinity capillary electrophoresis (PFACE) or affinity capillary electrophoresis (ACE). In these techniques, the column is first partially filled with increasing concentrations of D-Ala-D-Ala terminus peptides. This is followed by plugs of buffer, antibiotic and two noninteracting standards, and acetic and/or succinic anhydride (and buffer in the case of ACE). The order of the reagent plugs containing the antibiotic and anhydride varies with the charge of the glycopeptide. Upon electrophoresis, the antibiotic reacts with the anhydride yielding a derivative of Teic or Rist. Continued electrophoresis results in the overlap of the derivatized antibiotic and the plug of D-Ala-D-Ala peptide. Analysis of the change in the relative migration time ratio (RMTR) of the new glycopeptide relative to the standards, as a function of the concentration of the D-Ala-D-Ala ligand yields a value for the binding constant K(b). The techniques described here can be used to assess how the derivatization of drugs alters their affinities for target molecules. PMID- 12627442 TI - A comparison between electrokinetic capillary chromatography and absorption spectroscopy for the analysis of peptide-micelle association by weak hydrophobic interactions. AB - Micellar electrokinetic capillary chromatography (MEKC) was compared to absorption spectroscopy to estimate equilibrium association constans (K(as)) for peptide-micelle systems involving three peptides (leucine-enkephalin, methionine enkephalin and leucine-phenylalanine (LF)) and two surfactant micelles (sodium dodecyl sulfate (SDS) and cetyltrimethylammonium bromide (CTAB)). Buffer pH was chosen to minimize purely electrostatic interactions between peptides and micelles that could not be interrogated by absorption spectroscopy. Viscosity corrected MEKC mobilities gave reasonably similar estimates of K(as) between the two methods for all three peptide-SDS micelle systems, with K(as) values ranging from 13.7 +/- 0.3 to 49 +/- 1 M(-1). For CTAB, estimates of K(as) for LF-CTAB micelle association were of the same order of magnitude as the SDS micelle by the two methods of estimation. On the other hand, enkephalin-CTAB micelle binding was about 10 times stronger (K(as) = 122 +/- 3 M(-1) to 311 +/- 9 M(-1)) than the enkephalin-SDS micelle binding. In addition, MEKC underestimated the K(as) values relative to spectroscopy by a factor of 2-3 for the enkephalin-CTAB system. PMID- 12627443 TI - Separation of peptides and proteins by capillary electrophoresis using acidic buffers containing tetraalkylammonium cations and cyclodextrins. AB - A method for improving separations of peptides and other positively charged species in capillary zone electrophoresis with untreated capillaries using acidic buffers containing tetraalkylammonium cations is described. Tetramethylammonium and tetrabutylammonium cations dynamically modify the capillary surface, leading to a reversal in the direction of the electroosmotic flow. As a result, the adsorption of positively charged peptides and proteins is minimized, and resolution and peak capacity are improved as the migration of cationic analytes is counterbalanced by the electroosmotic flow. The combining effect of reversing electroosmotic flow and cyclodextrin inclusion complexation on separations of closely related peptides and a protein mixture, as well as tryptic digest of hemoglobin is demonstrated. PMID- 12627445 TI - Capillary electrophoresis-mass spectrometry of peptides from enzymatic protein hydrolysis: simulation and optimization. AB - Two important limitations still exist for the characterization of protein digests by capillary electrophoresis-mass spectrometry (CE-MS): (i) the buffer choice (i.e., the buffer must provide an adequate CE separation without ruining the MS signal), and (ii) the frequent generation of "unexpected" peptidic fragments during the enzymatic protein hydrolysis. In this work, a new approach is used to solve these difficulties, namely a theoretical model that relates the electrophoretic behavior of peptides to their sequence. The effectiveness of this procedure is demonstrated by the fast attainment of good CE-MS conditions for analyzing the peptides obtained from an enzymatic protein hydrolysate in a single run. This strategy can provide useful information for helping to characterize "unexpected" fragments from protein digests. PMID- 12627444 TI - Separation of anti-tumor peptides by capillary electrophoresis in organic solvent containing background electrolytes. AB - Connections between the calculated and measured electrophoretic mobilities (nu(ep)) determined by capillary electrophoresis as well as connections between the measured and calculated diffusion coefficients of anti-tumor peptides have been investigated in background electrolytes (BGEs) containing different organic solvents (acetonitrile, methanol, ethanol and isopropanol). Comparison of the electrophoretic mobility (nu(ep)) values revealed discrepancies between the measured and calculated values. However, no change in the migration order or selectivity could be expected from the calculated nu(ep) values, variation of both properties was observed applying organic solvents as BGE modifiers. Experimental determination of the diffusion coefficient suggested that the effect of the organic solvents is not restricted to the change of the BGE viscosity. The reason for the discrepancy between the measured and calculated mobility values might be the possible conformation and/or solvation changes of the peptide caused by the different organic solvents. PMID- 12627446 TI - Insoluble eggshell matrix proteins--their peptide mapping and partial characterization by capillary electrophoresis and high-performance liquid chromatography. AB - Avian eggshell matrix proteins were studied by two analytical approaches. Peptide mapping was done by trypsin and pepsin followed by collagenase cleavage; analyses were carried out by capillary electrophoresis and reversed-phase high-performance liquid chromatography (HPLC). Comparison of peptide maps obtained by both methods revealed a complex mixture of peptides in the insoluble layers of the eggshell; it was concluded that there are at least three different insoluble protein/peptide layers in the avian eggshell (cuticle, palisade, and mammillary layer). Partial characterization of peptides in each layer was made by HPLC-mass spectrometry analysis. There is an evidence that the eggshell insoluble proteins contain species susceptible to collagenase cleavage, however, the sequences split by this enzyme probably are not those typical for the main triple-helical core of collagenous proteins. It is proposed that the action of collagenase upon eggshell proteins is caused by the side effect of collagenase described previously with synthetic peptides. Some of the proteins present are probably glycosylated. Fatty acid content in the insoluble eggshell layers (after decalcification) was in the range of 2-4% (which reflected both lipid and lipoproteins bound fatty acids). Porphyrin pigments are dominant in the cuticle layer. PMID- 12627447 TI - Failure of immunocompetitive capillary electrophoresis assay to detect disease specific prion protein in buffy coat from humans and chimpanzees with Creutzfeldt Jakob disease. AB - The emergence of a new environmentally caused variant of Creutzfeldt-Jakob disease (vCJD), the result of food-born infection by the causative agent of bovine spongiform encephalopathy (BSE), has stimulated research on a practical diagnostic screening test. The immunocompetitive capillary electrophoresis (ICCE) assay has been reported to detect disease-specific, proteinase-resistant prion protein (PrPres) in the blood of scrapie-infected sheep. We have applied this method to blood from CJD-infected chimpanzees and humans. The threshold of detection achieved with our ICCE was 0.6 nM of synthetic peptide corresponding to the prion protein (PrP) C-terminus, and 2 nM of recombinant human PrP at the optimized conditions. However, the test was unable to distinguish between extracts of leucocytes from healthy and CJD-infected chimpanzees, and from healthy human donors and patients affected with various forms of CJD. Thus, the ICCE assay as presently performed is not suitable for use as a screening test in human transmissible spongiform encephalopathies (TSEs). PMID- 12627448 TI - Determination of egg white lysozyme by on-line coupled capillary isotachophoresis with capillary zone electrophoresis. AB - An on-line coupled capillary isotachophoresis - capillary zone electrophoresis method for the determination of lysozyme in selected food products is described. The optimized electrolyte system consisted of 10 mM NH(4)OH + 20 mM acetic acid (leading electrolyte), 5 mM epsilon -aminocaproic acid +5 mM acetic acid (terminating electrolyte), and 20 mM epsilon -aminocaproic acid +5 mM acetic acid +0.1% m/v hydroxypropylmethylcellulose (background electrolyte). A clear separation of lysozyme from other components of acidic sample extract was achieved within 15 min. Method characteristics, i.e., linearity (0-50 micrograms/mL), accuracy (recovery 96+/-5%), intra-assay (3.8%), quantification limit (1 microgram/ml), and detection limit (0.25 microgram/mL) were determined. Low laboriousness, sufficient sensitivity and low running costs are important attributes of this method. The developed method is suitable for the quantification of the egg content in egg pasta. PMID- 12627450 TI - Identification of degradation products of aspartyl tripeptides by capillary electrophoresis-tandem mass spectrometry. AB - Capillary electrophoresis-electrospray tandem mass spectrometry (CE-MS/MS) has been used to identify degradation products of the aspartyl tripeptides Phe-Asp GlyNH(2) and Gly-Asp-PheNH(2) following incubation of the peptides in acidic and alkaline solution. At pH 2, the dominant decomposition products resulted from cleavage of the peptide backbone amide bonds to yield the respective dipeptides and amino acids. In addition, the cyclic aspartyl succinimide intermediate was identified by its [M+H](+) at m/z = 319 and the MS/MS spectrum exhibiting a simple fragmentation pattern with the [C(8)H(10)N](+)-ion as the principal daughter ion (a(1) of Phe-Asp-GlyNH(2)). Deamidation of the C-terminal amide as well as isomerization and enantiomerization of the Asp residue occurred upon incubation at pH 10. alpha-Asp and the isomeric beta-Asp and most of the diastereomeric forms (corresponding to D/L-Asp) could be separated by CE. All isomers could be identified based on their MS/MS spectra. Peptides with the amino acid sequence Phe-Asp-Gly containing the regular alpha-Asp bond displayed a highly intense b(2) fragment ion and a low abundant y(2) ion. In contrast, the y(2) and a(1) fragment were high abundant daughter ions in the mass spectra of beta-Asp peptides while the b(2) ion exhibited a lower abundance. Differences in the MS/MS spectra of the isomers of the peptides with the sequence Gly-Asp-Phe were obvious but less pronounced. In conclusion, CE-MS/MS proved to be a useful tool to study the decomposition and enantiomerization of peptides including the isomerization of Asp residues, due to the combination of efficient separation of isomers by CE and their identification by MS/MS. PMID- 12627449 TI - Attomole sensitivity for unlabeled proteins and polypeptides with on-chip capillary electrophoresis and universal detection by interferometric backscatter. AB - A universal detector based on backscatter interferometry has been developed to perform nanoliter volume refractive index measurements for on-chip sodium dodecyl sulfate (SDS) gel based (polyethylene oxide gel) separations and quantification label-free proteins. The on-chip interferometric backscatter detector (OCIBD) system consists of a simple, folded optical train based on the interaction of a laser beam with an etched channel in the shape of half cylinder in a fused-silica plate. The backscattered light from the channel takes on the form of a high contrast interference pattern that contains information related to the bulk properties of the fluid located within the probe or detection volume of 2.32 x 10(-9) L. Depending on capillary electrophoresis (CE) injection method, the positional changes of the interference pattern extrema (fringes) allow for the quantification of unlabeled proteins at levels ranging from 11 to 310 amol (2.7 x 10(-8)mol/L) with a linear dynamic range of 2.5 decades (egg albumin). Using OCIBD microchannel-based SDS capillary gel electrophoresis (SDS/CGE), separation and detection of five label-free proteins was achieved in less than 100 seconds with detection limits ranging from 0.95 pg (1.1 x 10(-16)mol or 2.5 x 10( 7)mol/L) of calmodulin to 7.0 pg (1.0 x 10(-16)mol or 2.4 x 10(-7)mol/L) for bovine serum albumin (BSA) without signal filtering or active thermal control. This development shows that a universal detector based on backscatter interferometry can be used effectively for on-chip label-free solute analysis. PMID- 12627451 TI - Capillary electrophoresis coupled to time of flight-mass spectrometry of therapeutic peptide hormones. AB - We have established a method for separation and characterization of a series of peptide hormones of pharmaceutical interest and wide therapeutical use by capillary electrophoresis-electrospray-mass spectrometry (CE-ES-MS) using a sheath flow interface. Several parameters were systematically investigated, such as concentration of the electrolyte, organic solvent and sheath liquid composition, gas flow rates and capillary position. Moreover, limits of detection, linearity, repeatability and day-to-day reproducibility of the proposed method were studied in order to obtain the main quality parameters. PMID- 12627452 TI - Analysis of the performance of antibody capture methods using fluorescent peptides with capillary zone electrophoresis with laser-induced fluorescence. AB - A method to analyze the performance of an antibody capture method using fluorescent peptides by capillary zone electrophoresis using laser-induced fluorescence (CZE-LIF) for detection has been developed. Fluorescent peptides from the prion protein were synthesized and the corresponding antibodies were produced in rabbits against these peptides. The antibodies were used to capture the fluorescent peptides. The antibodies were then bound to protein A Sepharose. After elution, the amount of fluorescent peptide that was captured vs. the total amount placed in the assay was evaluated by CZE-LIF. Of the three peptides used in this evaluation, it was found that the recovery was approximately 25-35%. When the abnormal prion protein was prepared from scrapie-infected brain samples from hamsters and a sheep using the previously described extraction method and this method, the amount of abnormal prion protein that was measured in the fluorescence immunoassay correlated with amounts estimated from Western blot. We conclude that this method can be used to detect abnormal prion protein in a tissue sample. PMID- 12627453 TI - On-line coupling of size exclusion and capillary zone electrophoresis via a reversed-phase C18 trapping column for the analysis of structurally related enkephalins in cerebrospinal fluid. AB - On-line coupled analytical techniques can be advantageous in the assay of smaller peptides in complex biological matrices such as plasma, cerebrospinal fluid (CSF) and tissues. The present study shows the feasibility of a recently developed system, consisting of a size-exclusion chromatographic (SEC) separation followed by a trapping procedure on an RP18 microcolumn with subsequent elution of the trapped fraction and separation by capillary zone electrophoresis (CZE) for the quantification of structural-related peptides in biological matrices, as demonstrated for a number of enkephalins in CSF. After SEC separation of the enkephalins from large proteins present in CSF a heart-cut of 200 nuL, containing the enkephalin peak, is taken, concentrated on the RP18 microcolumn and, after elution of the enkephalins with 80% acetonitrile, a fraction of the eluate is electrokinetically injected into the CZE system, where stacking and separation is achieved. The degradation of the peptides, caused by endogenous peptidases in the matrix, is sufficiently inhibited with imipramine HCl. The assay has a satisfactory linearity and intraday (9.70-16.3%) precision considering the complexity of this multidimensional separation system. The sensitivity of the method, with a concentration limit of quantification of 2.5 nug/mL, is comparable with other CZE assays for peptides and sufficient for the quantification of peptide drugs in biological matrices. PMID- 12627454 TI - A glycopeptide antibiotic chiral stationary phase for the enantiomer resolution of hydroxy acid derivatives by capillary electrochromatography. AB - Separation of hydroxy acid enantiomers was achieved by using capillary electrochromatography (CEC) employing a chiral stationary phase (CSP) based on MDL 63,246 (Hepta-Tyr), a macrocyclic antibiotic of the teicoplanin family. The chiral selector was chemically bonded to 5 num diol-modified silica particles and the CSP mixed with amino silica (3:1 w/w) was packed into a 75 num ID fused silica capillary. The CEC experiments were carried out by using an aqueous reversed-phase mode for the enantiomeric resolution of hydroxy acid compounds. Good enantioresolution was achieved for mandelic acid (MA), m-hydroxymandelic acid (m-OH-MA), p-OH-MA, and 3-hydroxy-4-methoxymandelic acid (3-OH-4-MeO-MA). The CEC system was less enantioselective towards 2-phenyllactic acid (2-PhL) and 3-PhL while mandelic acid methyl ester (MA-Et-Est) enantiomers were not resolved. Several experimental parameters, such as organic solvent type and concentration, buffer pH, capillary temperature, on enantioresolution factor, retention time, and retention factor were studied. PMID- 12627456 TI - NOTCH4 and the frontal lobe in schizophrenia. AB - NOTCH4 is a developmentally expressed gene recently reported to be in linkage disequilibrium (LD) with schizophrenia. We investigated this finding in our sample of subjects, focusing on an exonic (CTG)(n) polymorphism, examining not only the association of this polymorphism with the disease phenotype, but also its effect on frontal lobe brain morphology and cognitive function in both affected individuals and a psychiatrically normal comparison group. While we did not find any association or LD with schizophrenia, we identified striking effects of NOTCH4 variability on the trait measures. Within the respective schizophrenia and comparison groups, NOTCH4 allelic variability was correlated with differences in measures of frontal lobe cognitive performance and frontal lobe brain tissue volumes that were intuitively congruent. These within-group effects, however, were in opposite directions across groups. These findings may reflect the interaction of NOTCH4 with the underlying genetic and phenotypic complexity that characterizes both schizophrenia and normal cognition and brain development. PMID- 12627457 TI - Modest evidence for linkage and possible confirmation of association between NOTCH4 and schizophrenia in a large Veterans Affairs Cooperative Study sample. AB - Wei and Hemmings [2000: Nat Genet 25:376-377], using 80 British parent-offspring trios, identified a number of NOTCH4 variants and haplotypes that showed statistically significant evidence of association to schizophrenia. Specifically, the 10 repeat allele of a (CTG)(n) marker and the 8 repeat allele of a (TAA)(n) marker demonstrated excess transmission to affected individuals; SNP21 and haplotypes SNP2-(CTG)(n) and SNP12-SNP2-(CTG)(n) also showed significant associations. In an attempt to replicate these findings, we tested for linkage and association between the same five markers used by Wei and Hemmings in 166 families collected from a multi-center study conducted by the Department of Veterans Affairs (DVA) Cooperative Study Program (CSP). The families include 392 affected subjects (schizophrenia or schizoaffective disorder, depressed) and 216 affected sibling pairs. The families represent a mix of European Americans (n = 62, 37%), African Americans (n = 60, 36%), and racially mixed or other races (n = 44, 27%). We identified moderate evidence for linkage in the pooled race sample (LOD = 1.25) and found excess transmission of the 8 (P = 0.06) and 13 (P = 0.04) repeat alleles of the (TAA)(n) marker to African American schizophrenic subjects. The 8 and 13 repeat alleles were previously identified to be positively associated with schizophrenia by Wei and Hemmings [2000: Nat Genet 25:376-377] and Sklar et al. [2001: Nat Genet 28:126-128], respectively. PMID- 12627458 TI - Association analysis of the HOPA12bp polymorphism in schizophrenia and manic depressive illness. AB - Variations in exon 42 of the HOPA (human opposite paired) gene have been associated with mental retardation, hypothyroidism and psychiatric disorders. We attempted to replicate the association with schizophrenia using 309 parent offspring trios from Bulgaria and 367 unrelated cases and 368 blood donors from the UK. We also tested 125 bipolar trios from Bulgaria, 112 bipolar trios from the UK and a sample of 178 unrelated bipolar cases and 188 blood donors from the UK. The frequency of HOPA(12bp) in the 556 UK blood donors was 2.6% and it was not significantly different in the UK patients groups, where it ranged from 1.2 to 3.8%. Sixteen mothers transmitted the HOPA(12bp) allele to schizophrenic offspring, while 12 did not transmit, a non-significant difference. There was a trend for under-transmission of the rare allele to bipolar patients (T/NT = 4/10) and they had a lower rate of that allele than schizophrenic patients in the Bulgarian population (1% vs. 4.2%, P = 0.043). However the two diagnostic groups had similar allele frequencies in the UK populations: 2% versus 2.6%, P = 0.6. We conclude that the HOPA polymorphism is unlikely to be a major risk factor in the pathogenesis of these major psychiatric disorders although there could be a small effect in schizophrenia. PMID- 12627459 TI - Distribution of haplotypes derived from three common variants of the NR4A2 gene in Japanese patients with schizophrenia. AB - Dysregulation in dopaminergic neurotransmission might play a role in the pathogenesis of schizophrenia, and therefore genetic components of the dopamine (DA) pathway may confer risk. The NR4A2 (Nurr1) gene is essential for the development and maintenance of mesencephalic DA-synthesizing neurons. Moreover, Nurr1 forms a heterodimer with the retinoid X receptor and disturbances in the retinoid-signaling cascade may be involved in susceptibility to schizophrenia. To investigate the potential genetic contribution of NR4A2, we performed a case control association study using three common variants in the gene [-2922(C)2-3, IVS6 + 17 approximately +18insG, EX8 + 657(CA)9-10] that were in strong linkage disequilibrium with each other. We did not detect a significant allelic or genotypic association. Haplotypes derived from all three polymorphisms generated similar results. These data do not support the notion that the NR4A2 gene plays a major role in risk for schizophrenia among Japanese individuals. PMID- 12627460 TI - Is there a phenotypic difference between probands in case-control versus family based association studies? PMID- 12627462 TI - Association between the CCK-A receptor gene and panic disorder. AB - Cholecystokinin (CCK) is one of the most abundant neurotransmitter peptides in the brain. CCK appears to play an important role in the neurobiology of anxiety and panic disorders (PD) in both humans and animals. Recently, we reported that lack of CCKAR had a significant anxiogenic-like effect in rats. In this study, to investigate the role of CCKAR in PD, we compared the CCKAR gene in PD patients and normal controls. Subjects who fulfilled the DSM-IV criteria for PD were 17 males and 26 females. The sequence containing the Pst I polymorphic site in the boundary between intron 1 and exon 2 of the CCKAR gene was studied. Pst I digestion of the PCR products gave two individual alleles: A1 and A2. The A1 allele was the undigested fragment and the A2 allele was the digested one with two variant bands at 264 and 180 bp. Genotypic frequencies were 20.9% (A1-A1), 55.8% (A1-A2), and 41.7% (A2-A2) in patients, and 20.5% (A1-A1), 46.2% (A1-A2), and 33.3% (A2-A2) in controls. Allelic frequencies were 48.8% (A1) and 51.2% (A2) in patients, and 43.6% (A1) and 56.4% (A2) in controls. The chi-square test did not show a significant difference in either genotypic or allelic frequencies between patients and control subjects. The Pst polymorphism of CCKAR may not be associated with PD. PMID- 12627463 TI - Transcriptional activities of cholecystokinin promoter haplotypes and their relevance to panic disorder susceptibility. AB - We previously identified a polymorphic compound short tandem repeat (STR) in the 5'-regulatory region of the cholecystokinin (CCK) gene, and showed that when the STRs were classified into three groups based on length and linkage disequilibrium behavior with neighboring variants, the medium class allele was significantly associated with panic disorder. The present study examined the transcriptional activity of the CCK promoter construct containing the STR and downstream -188A/G variation. The STRs acted as transcriptional repressors with a similar potency among the three classes, but the long (L) class STR exhibited a synergistic effect on decreasing promoter activity when combined with -188G. The haplotype composed of the L class of STR and -188G was significantly less frequent in panic disorder (P = 0.0032; odds ratio, 95% confidence interval = 0.06, 0.01-0.69). These results suggest that the L-(-188G) haplotype may act as a protective factor against panic by reducing the expression of anxiogenic CCK. PMID- 12627464 TI - Suicide and serotonin: study of variation at seven serotonin receptor genes in suicide completers. AB - Suicide is an important public health problem, accounting for a significant proportion of total mortality among young people, particularly males. There is growing and consistent evidence suggesting that genetic factors play an important role in the predisposition to suicide. Based on several lines of evidence supporting a reduced serotonergic neurotransmission in subjects who committed suicide, we investigated variation at genes that code for serotonin receptor 1B (5-HTR1B), 1Dalpha (5-HTR1Dalpha), 1E (5-HTR1E), 1F (5-HTR1F), 2C (5-HTR2C), 5A (5-HTR5A), and 6 (5-HTR6) in a total sample of 106 suicide completers and 120 normal controls. No differences were observed in allelic or genotypic distributions between groups for any of the loci investigated. Moreover, further analysis according to suicide method or psychopathology also failed to reveal differences between groups. Our results do not support a substantial role of these serotonergic receptors in suicide completion. PMID- 12627465 TI - Allelic variation in serotonin transporter function associated with the intensity dependence of the auditory evoked potential. AB - The intensity dependence of the auditory evoked potential (AEP) has been suggested as an indicator of central serotonergic function, a strong intensity dependence presumably reflecting low serotonergic activity. As individual differences in serotonergic neurotransmission can be accounted for in part by genetic variation in genes of the serotonergic pathway, we investigated whether a functional polymorphism in the promoter region of the serotonin transporter gene (5-HTTLPR) is associated with the AEP intensity dependence. Because dopaminergic influences on the intensity dependence have also been reported, we furthermore explored the role of a functional polymorphism in the dopamine D4 receptor gene (DRD4 exon III) in the modulation of the AEP intensity dependence. AEPs to tones of six intensity levels were recorded from 60 healthy young individuals, and N1/P2 linear as well as median slopes at central electrode sites were computed as measures of the AEP intensity dependence. Analyses of variance showed that there was a significant effect of the 5-HTTLPR on the AEP intensity dependence. Individuals with the ll genotype exhibited a stronger intensity dependence compared to individuals with the ls genotype. This effect was even more pronounced when DRD4 exon III was considered in the analyses. In conclusion, these findings provide further evidence for a role of serotonergic neurotransmission in the modulation of the AEP intensity dependence. The results also point to possible dopaminergic influences on the AEP intensity dependence. PMID- 12627466 TI - Elucidating the role of genetic factors in smoking behavior and nicotine dependence. AB - Despite over a decade of intensive tobacco control efforts, a substantial proportion of the world's population continues to smoke. With advances in molecular biology and genomics technology, attention is focusing on the genetic basis of addiction to nicotine. The hope is that by better understanding the biology of nicotine addiction, novel prevention and cessation strategies can be developed and targeted to individuals most likely to benefit. This article provides an overview of currently available data from genetics research on smoking behavior and discusses study design issues to be addressed to make progress in this line of research. PMID- 12627467 TI - Gender-specific molecular heterosis and association studies: dopamine D2 receptor gene and smoking. AB - If the concept of the gender-specific molecular heterosis is not considered and tested, incorrect conclusions would easily be drawn in association studies. Therein, heterosis and its gender effect in the genetic effect of DRD2 gene for smoking were examined with 187 healthy Korean individuals. The male smokers showed a higher A1 allele frequency (P = 0.016) and prevalence (P = 0.049) than those of the male non-smokers, and the female smokers showed a lower frequency of heterozygotes (P = 0.018) than the female non-smokers. However, the association of DRD2 gene with smoking found in each gender disappeared when both males and females were considered as one group because of the opposite genetic effect of DRD2 gene for smoking: (1) while 75% of heterozygotes males were smokers, only 22% of female heterozygotes were; (2) males showed an excess of heterozygotes and the deviation from Hardy-Weinberg expectations in smokers, while these were true in the female non-smokers; and (3) in non-smokers, females were different from the males exhibiting a significantly higher prevalence (P = 0.005) and frequency (P = 0.015) of A1 allele, and significantly different genotype (P = 0.017) distribution, and higher frequency of heterozygotes (P = 0.055). Meanwhile, in smokers, males showed higher frequency of heterozygotes (P = 0.019) compared to females. The results indicate that gender-specific molecular heterosis at DRD2 gene for smoking is also applicable in healthy individuals as well as schizophrenics. Moreover, this concept has general applicability to other candidate genes and biological phenotypes. PMID- 12627468 TI - Association between the cortisol response to opioid blockade and the Asn40Asp polymorphism at the mu-opioid receptor locus (OPRM1). AB - This study examined whether a reportedly functional polymorphism in the gene encoding the mu-opioid receptor protein (A118G, which causes an Asn40Asp substitution in the receptor's extracellular domain), modifies the cortisol response to the opioid antagonist naloxone. The polymorphism occurs commonly in European Americans and some other population groups, underscoring its potential phenotypic significance. METHODS: Using a balanced, within-subject design involving two test sessions over a period of 3-7 days, we examined ACTH and cortisol responses to intravenous naloxone (125 microg/kg) or placebo in 30 healthy subjects (21 males, mean age = 24.4 years). Plasma ACTH and cortisol concentrations were measured over 120 min post infusion. DNA isolated from whole blood was PCR amplified and genotyped via restriction enzyme digestion, with genotypes assigned based on agarose gel size fractionation. RESULTS: Subjects with one or more Asp40 alleles (n = 6; 5 heterozygotes and 1 homozygote) had significantly higher cortisol concentrations at baseline and at 15, 60, and 90 min after naloxone infusion than subjects homozygous for the Asn40 allele (n = 24). Subjects with the Asp40 allele also had a greater peak cortisol response and a greater area under the cortisol time curve than those homozygous for the Asn40 allele. There were no effects of the Asn40Asp polymorphism on plasma ACTH concentration or on self-reported anxiety or distress. CONCLUSIONS: These findings are consistent with recent reports showing an enhanced cortisol response to naloxone and a reduced agonist effect of morphine-6-glucuronide among subjects with the Asp40 variant. Given evidence of its pharmacological significance, the clinical relevance of this polymorphism warrants further investigation. PMID- 12627469 TI - Linkage mapping of beta 2 EEG waves via non-parametric regression. AB - Parametric linkage methods for analyzing quantitative trait loci are sensitive to violations in trait distributional assumptions. Non-parametric methods are relatively more robust. In this article, we modify the non-parametric regression procedure proposed by Ghosh and Majumder [2000: Am J Hum Genet 66:1046-1061] to map Beta 2 EEG waves using genome-wide data generated in the COGA project. Significant linkage findings are obtained on chromosomes 1, 4, 5, and 15 with findings at multiple regions on chromosomes 4 and 15. We analyze the data both with and without incorporating alcoholism as a covariate. We also test for epistatic interactions between regions of the genome exhibiting significant linkage with the EEG phenotypes and find evidence of epistatic interactions between a region each on chromosome 1 and chromosome 4 with one region on chromosome 15. While regressing out the effect of alcoholism does not affect the linkage findings, the epistatic interactions become statistically insignificant. PMID- 12627470 TI - Variation in the coding sequence and flanking splice junctions of the estrogen receptor alpha (ERalpha) gene does not play an important role in genetic susceptibility to bipolar disorder or bipolar affective puerperal psychosis. AB - Genes involved in estrogen pathways have been proposed as possible candidates influencing susceptibility to bipolar disorder and the affective symptoms suffered by many women during the puerperal period. The estrogen receptor alpha (ERalpha) gene in particular has been a subject of interest and has recently been intensively screened for variations of potential relevance to psychiatric disorders, resulting in the identification of four mutations in individuals with bipolar disorder or puerperal psychosis. We have examined the frequency of these four ERalpha variations in a case control study using a group of mixed gender bipolar individuals (N = 231), further classified into subsets of parous bipolar females with (N = 112) and without (N = 50) puerperal psychosis, and a non psychiatric comparison group (N = 110). We have also investigated the families in which the variations were initially detected, for evidence of co-segregation of the variants with mood disorder. We found no evidence in our case control sample to support the involvement of any of the ERalpha variations in either the aetiology of bipolar disorder or puerperal triggering of bipolar episodes. Nor did we find co-segregation of ERalpha variants and disease in any of the four families examined. We conclude that variation in the coding sequence and flanking splice junctions of the ERalpha gene does not play an important pathogenic role in the majority of cases of Bipolar Disorder or Bipolar Affective Puerperal Psychosis. PMID- 12627471 TI - Allelic variants interaction of dopamine receptor D4 polymorphism correlate with personality traits in young Korean female population. AB - Polymorphism in exon III of the dopamine D4 receptor (DRD4) gene has been implicated to be associated with the human personality trait of novelty seeking (NS). For this study, we have investigated the possible association between 48-bp VNTR in exon III and -521 C/T SNP of the DRD4 and personality traits among young ( approximately 14 years of age) Korean female population. We found that the interaction between the two alleles of DRD4 polymorphism, 48-bp VNTR and -521 C/T, were significantly high on NS (F = 4.88, P = 0.029) and persistence (P) (F = 5.05, P = 0.027) personality scores, suggesting that the variants of DRD4 gene influence the NS and P (persistent) personality traits. When analyzed independently, however, the two different alleles of DRD4 polymorphisms, 48-bp VNTR and -521 C/T, there was no direct correlation with the personality traits. PMID- 12627472 TI - White matter tract alterations in fragile X syndrome: preliminary evidence from diffusion tensor imaging. AB - Fragile X syndrome, the most common form of hereditary mental retardation, causes disruption in the development of dendrites and synapses, the targets for axonal growth in the central nervous system. This disruption could potentially affect the development, wiring, and targeting of axons. The current study utilized diffusion tensor imaging (DTI) to investigate whether white matter tract integrity and connectivity are altered in fragile X syndrome. Ten females with a diagnosis of fragile X syndrome and ten, age matched, female control subjects underwent diffusion weighted MRI scans. A whole brain analysis of fractional anisotropy (FA) values was performed using statistical parametric mapping (SPM). A follow-up, regions-of-interest analysis also was conducted. Relative to controls, females with fragile X exhibited lower FA values in white matter in fronto-striatal pathways, as well as in parietal sensory-motor tracts. This preliminary study suggests that regionally specific alterations of white matter integrity occur in females with fragile X. Aberrant white matter connectivity in these regions is consistent with the profile of cognitive and behavioral features of fragile X syndrome, and potentially provide additional insight into the detrimental effects of suboptimal levels of FMRP in the developing brain. PMID- 12627473 TI - Continuing the search for dyslexia genes on 6p. AB - This study is a continuation and extension of the work with Orton Developmental Dyslexia (DD) pedigrees [Grigorenko et al., 1997; 2000, Grigorenko et al., 2001]. This study utilized an extended sample (N = 176) and a well-saturated map of chromosome 6p (30 markers). Six phenotypes were constructed to span a range of dyslexia-related cognitive processes. These phenotypes were: (1) Phonemic Awareness (of spoken words); (2) Phonological Decoding (of printed nonwords); (3) Rapid Automatized Naming (of colored squares or object drawings); (4) Single Word Reading (orally, of printed real words); (5) Phonemic Awareness/Decoding/Single Word Reading pathway; and (6) Phonemic Awareness/Rapid Naming/Single-Word Reading pathway. The study resulted in two major findings. First, considering the distributions of the genetic linkage indicators across all phenotypes examined, there appear to be three regions of interest (around markers D6S109, D6S1261, and in the D6S105-D6S265 region). Any of these regions could serve as a starting point in the search for specific gene candidates contributing to the manifestation of DD, yet they all might be echo peaks of a single peak, the boundary of which is difficult to establish due to the limited power of this sample. Second, the DD-related linkage in 6p21.3 appears to be most closely related to the manifestations of DD through phonemic awareness and single-word reading deficits. PMID- 12627474 TI - Enhanced production of amyloid precursor protein mRNA by peripheral mononuclear blood cell in Alzheimer's disease. AB - Previous studies have suggested the involvement of amyloid precursor protein (APP) in Alzheimer's disease (AD), as exons 16 and 17 of the APP gene mutations have been found in some familial AD patients. Furthermore, overexpression and deposition of the beta amyloid peptide, a proteolytic product of APP, have been considered as a pathological hallmark of Alzheimer's disease. Therefore, it is of particular interest to determine the expression of APP gene at the transcription level for better understanding of the roles of APP gene in AD pathogenesis. In this work, we employed the quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) to quantify APP mRNA transcripts in the peripheral mononuclear blood cells (PMBC) of 52 Alzheimer's patients, 28 vascular dementia (VD) patients, and 60 healthy elderly controls. The results showed that the amount (mean +/- SEM) of APP transcripts per microgram of total cDNA was 4.05 +/- 0.27, 2.73 +/- 0.33, and 2.59 +/- 0.27 amole in AD, VD, and healthy controls, respectively. There was a significant increase (P < 0.05) in the expression of APP mRNA transcripts in AD compared with that in VD or in healthy controls. Thus, our data indicated that variation of APP gene expression in PMBC might be a pathogenic source of Alzheimer's disease. PMID- 12627476 TI - A "line item" approach to the identification of genes involved in polygenic behavioral disorders: the adrenergic alpha2A (ADRA2A) gene. AB - The usual approach for using single base pair polymorphisms (SNPs) for the investigation of the genetics of behavioral disorders is to examine a single diagnostic syndrome or personality trait based on variables relating to a cluster of behavioral symptoms. However, since some of these variables may address behaviors that are associated with one allele while others are associated with the other allele, the overall association may be non-significant and significant sub-syndromal associations may be missed. Thus, we have reversed the process in a technique we term a "line item" approach. As a test of the technique we have examined the association between genotypes of a C- > G-1291 Msp I promotor SNP of the ADRA2A gene and 390 individual symptoms from a structured review of DSM-IV criteria for twelve different groups of symptoms. We examined 334 individuals consisting of controls and subjects with Tourette syndrome (TS). Based on the mean scores for each genotype, those symptoms that were individually significant at alpha < or = 0.05 fell into three major groups by mode of inheritance: allele 1 codominant (11 > 12 > 22), allele 2 codominant (22 > 12 > 11), and negative heterosis (12 < 11, 22). Within each mode of inheritance group, the number of symptoms that were significant for the twelve symptom clusters was compared by chi-square analysis. This showed that symptoms were drawn from the diagnostic groups in a significantly non-random fashion. Thus, the allele 1 codominant symptoms came from the anxiety, affective, schizoid, and somatization diagnostic groups (internalizing symptoms) (chi(2) = 80.0, d.f. = 11, P < or = 0.0000001), while the allele 2 codominant symptoms came from the ADHD and oppositional defiant/conduct disorder diagnostic groups (externalizing symptoms) (chi(2) = 81.0, d.f. = 11, P < or = 0.0000001). The questions that fell in the negative heterosis type of inheritance were not significantly associated with specific diagnostic groups (P = 0.87). These results showed that the ADRA2A gene was associated with symptoms of autonomic, sympathetic dysfunction from different diagnostic groups. The advantages of the "line item" approach include (a) the identification of the symptoms associated with each allele, (b) the identification of symptom clusters independent of DSM diagnoses, (c) the elucidation of heterosis and other mode of inheritance effects, (d) the distinction between an association with a primary disorder versus a comorbid disorder, (e) the identification of associations with sub-syndromal symptom clusters that do meet full DSM-IV criteria, and (f) the identification of symptom clusters across databases. PMID- 12627477 TI - Foreword: The Fourth International Conference on Varicella, Herpes Zoster and Post-Herpetic Neuralgia (PHN). PMID- 12627475 TI - Family-based and case-control association studies of catechol-O-methyltransferase in attention deficit hyperactivity disorder suggest genetic sexual dimorphism. AB - Attention deficit hyperactivity disorder (ADHD) is the most common childhood onset behavioral disorder. Boys are more often affected than girls. Family, twin, and adoption studies have supported a strong genetic basis. Some studies show that a catechol-O-methyltransferase (COMT) polymorphism affecting enzyme activity was associated with personality characteristics and diseases, such as novelty seeking personality, substance abuse, and heroin addiction, whose features are similar to ADHD or are associated with ADHD. These findings suggest that the COMT gene may be a candidate gene for ADHD. TDT, HHRR, and case-control association studies were conducted within a sample of 202 nuclear ADHD families, 340 ADHD cases, and 226 controls in the Han Chinese population. Diagnoses and ADHD subtypes were ascertained according to DSM-IV criteria using American Clinical Diagnostic Interviewing Scales. The HHRR analysis suggested that the low enzyme activity COMT Met allele was preferentially transmitted to ADHD boys (160 trios, chi(2) = 3.858, P = 0.05, df = 1) but not girls. This association is particularly pronounced among male ADHD probands without any comorbidity (50 trios, HHRR: chi(2) = 5.128, P = 0.024, df = 1; TDT: chi(2) = 4.558, P = 0.033, df = 1), especially the ADHD-I subtype (32 trios, HHRR: chi(2) = 5.792, P = 0.016, df = 1; TDT: chi(2) = 5.333, P = 0.021, df = 1). The case-control study revealed that the Val allele was more frequent in females meeting ICD-10 or DSM-IV criteria for ADHD than in female controls (86 and 79.5%, respectively, chi(2) = 4.059, P = 0.044, df = 1). Although these results suggest the COMT gene exerts some influence on the risk for ADHD in the Han Chinese population, given the potential for Type I error, these findings require replication before drawing definitive conclusions. PMID- 12627479 TI - Pathogenesis of simian varicella virus. AB - Simian varicella virus (SVV) is closely related to varicella-zoster virus (VZV) and induces a natural varicella-like disease in nonhuman primates. Therefore, simian varicella is a useful model to investigate varicella pathogenesis and to evaluate antiviral therapies. In this report, we review recent studies on SVV pathogenesis and latency. Experimental infection of African green monkeys is followed by a 7-10 day incubation period during which a viremia disseminates the virus throughout the body. Clinical disease is characterized by fever and vesicular skin rash. Pneumonia and hepatitis may occur during more severe infections. Examination of acutely infected tissues reveals histopathology including necrosis and hemorrhage in the skin, lung, liver, and spleen. In contrast, the neural ganglia exhibit minimal histopathology. SVV DNA, immediate early, early, and late gene transcripts, and viral antigens are detected in the tissues of acutely infected monkeys. Host immune responses are induced which resolve the acute infection within 21 days. During or after acute infection, SVV establishes latent infection in the ganglia of surviving monkeys. The virus may reactivate later in life to cause secondary disease and viral transmission to susceptible monkeys. PMID- 12627480 TI - Epidemiology of Varicella-Zoster Virus in England and Wales. AB - Many countries are studying currently the possibility of mass vaccination against varicella. The objective of this study was to provide a complete picture of the pre-vaccine epidemiology of the Varicella-Zoster Virus in England and Wales to aid in the design of immunisation programs. Population-based data including general practitioner sentinel surveillance, hospitalisation data, and death certificates from England and Wales were analysed. The average incidence rates for varicella and zoster between 1991 and 2000 were 1,291 and 373 per 100,000 years, respectively. Overall hospitalisation rates were equal for varicella and zoster (4.5 vs. 4.4 hospitalisation per 100,000 population) with 5 and 8%, respectively, having underlying immunosuppressive conditions. The age-specific proportion of cases hospitalised and length of stay were similar between the two diseases. However, the overall burden of disease is considerably higher for zoster. The number of inpatient days and case-fatality due to zoster are roughly 4 to 6 times greater than for varicella (11 vs. 3 days and 25 vs. 4 deaths per 100,000 case). These results provide base-line estimates should mass varicella vaccination be introduced in England and Wales. PMID- 12627481 TI - Varicella susceptibility and vaccine use among young adults enlisting in the United States Navy. AB - Primary varicella infection, or chicken pox, is a threat to all young adults who join the United States (U.S.) military if they fail to develop immunity prior to enlistment. Historically, outbreaks of chicken pox have caused marked morbidity and impaired military readiness. In December 1996, the U.S. Navy began performing serologic testing for varicella among all new recruits, and vaccinating those found to be sero-negative. We evaluated results of the screening program in its first 4 years, and used multivariable logistic regression modeling to describe factors associated with varicella susceptibility. Cases of chicken pox were tracked among all military services before and after program implementation. More than 190,000 young adults enlisted in the U.S. Navy between 1997 and 2000. Recruits originated from all 50 states and several foreign countries; 84% were male, and their average age was 19 years. Seven percent were found to be susceptible (sero-negative) to varicella. In multivariable modeling, race/ethnicity was associated with susceptibility, but age, gender, and home state were not. The overall incidence of chicken pox in the Navy was reduced by more than 80% after initiation of the screening-vaccination program. A successful varicella screening-vaccination program has been implemented in the U.S. Navy. Results of serologic screening undertaken on this large number of young adults may be useful in tracking the changing epidemiology of varicella in the general population in the post-vaccine era. PMID- 12627482 TI - Geographic and racial aspects of herpes zoster. AB - Geographic and racial factors have been reported in studies of the epidemiology of varicella and herpes zoster. To clarify further these relationships, data from five multicenter clinical trials of the antiviral agent famciclovir were examined (total N = 2074). Non-Caucasian racial group and tropical region were each significantly associated with younger age at zoster onset. In analyses of the non Caucasian subgroups, Black and Asian patients did not significantly differ in age or sex; however, Black and Asian patients from tropical regions had significantly younger mean ages at onset and greater rash duration at enrollment than those from temperate regions. Controlling for sex and rash duration at enrollment, both tropical region and non-Caucasian racial group were found to be independently associated with a younger age at zoster onset. These results suggest that racial group and geographic region may be independent factors associated with age at onset in patients with herpes zoster. PMID- 12627483 TI - A study of shingles and the development of postherpetic neuralgia in East London. AB - The incidence of post-herpetic neuralgia following shingles and the factors that are known to predict it were examined in a prospective observational community study of patients with acute shingles presenting to their family doctors. The detection of viral DNA in the blood at presentation as a prognostic indicator for pain was also evaluated. Patients were followed for one year and the persistence of pain following rash assessed. Among 165 patients who had completed 6 months, and 139 one-year follow-up, the prevalence of post herpetic neuralgia was 30% at 6 weeks 27% at 12 weeks, 15.9% at 6 months, and 9% at one year. Age and severity of pain were significantly associated with the persistence of pain beyond 3 months. Viremia at presentation was detected in 66% of patients and was significantly associated with the presence of pain at six months or beyond. Antiviral agents were administered to only 50% of those at highest risk of post herpetic neuralgia (PHN) mainly because of presentation longer than 72 hours after the onset of rash. Few patients were prescribed the more potent prodrugs, Valaciclovir and Famciclovir. In conclusion, treatment of acute shingles in this observational community-based study was suboptimal in 50% of cases. More accurate prediction of which subset of elderly patients are most at risk of PHN may enable targeted prescribing of the most potent drugs to those most likely to benefit. PMID- 12627484 TI - Varicella vaccination: impact of vaccine efficacy on the epidemiology of VZV. AB - In 1995, varicella vaccination was introduced into the infant immunization schedule of the United States. Currently, many other countries are considering mass varicella vaccination. Mass vaccination has two dangers: it could increase the number of varicella cases in adults, where severity is greater, and increase cases of zoster. A deterministic, realistic, age-structured model (RAS) was built to study these concerns. Model parameter estimates were derived from a review of the literature and surveillance data from England and Wales. Different vaccine efficacy scenarios, vaccine coverages, and vaccination strategies were investigated. The model predicts that, although an upward shift in the age at infection occurs, the overall morbidity due to varicella is likely to decrease following mass infant vaccination. On the other hand, cases of zoster may significantly increase in the first 50 years following vaccination. The model predicts that, in a population similar to England and Wales (50 m people), varicella vaccination with 90% coverage would prevent 0.6 m inpatient days due to varicella but would generate an extra 1.1 m inpatient days due to zoster over the first 65 years. Thus, under base-case model assumptions, the gain in reduction of varicella morbidity from infant vaccination is offset in the short-term by the increases in zoster morbidity (using inpatient days as a proxy). Paradoxically, less effective vaccines or vaccine programmes can be more effective in reducing overall morbidity (varicella + zoster) by allowing the virus to circulate more, which produces a smaller shift in the age at infection and a smaller increase in zoster cases. PMID- 12627485 TI - Measurement of cell-mediated immunity with a Varicella-Zoster Virus-specific interferon-gamma ELISPOT assay: responses in an elderly population receiving a booster immunization. AB - An interferon-gamma ELISPOT assay has been developed for assessment of cellular immune responses to Varicella-Zoster Virus (VZV) in large, multi-center clinical vaccine trials. We show that the assay performed best when testing peripheral blood mononuclear cells (PBMCs) that had been isolated and then frozen on the same day as blood was drawn, and that freezing PBMCs from blood that was stored overnight before processing resulted in dramatically reduced responses. This assay was used to monitor cell-mediated immunity (CMI) in response to a booster immunization with an investigational live, attenuated VZV vaccine in an elderly population that had been vaccinated 8-10 years previously. The booster vaccine elicited a 1.6- to 1.7-fold rise in the VZV-specific cellular immune response as measured by the ELISPOT assay. The increase from pre to post booster vaccination response was more pronounced (approximately 2.2-fold rise) in a subset of subjects who had received two prior immunizations with a live, attenuated vaccine. PMID- 12627486 TI - Genetic variation of varicella-zoster virus: evidence for geographical separation of strains. AB - Heteroduplex mobility assay was used to identify variants of varicella-zoster virus (VZV) circulating in the United Kingdom and elsewhere. Forty variable positions were identified. Sixteen substitutions were non-synonymous, resulting in an amino acid change, the majority of which were clustered within surface expressed proteins. Phylogenetic analysis distinguished at least three major clades (strains A, B, C) supported by significant bootstrap values. Apart from the United Kingdom and Brazil where all three strains were found, genotypes appeared to be closely associated with the geographical region in which they were sampled. Allelic co-segregation of widely spaced single nucleotide polymorphisms (SNPs) confirmed the genetic stability of the VZV. Recombination rates were difficult to calculate because of the low intra genotypic variation. However, one haplotype originating from Brazil is most parsimoniously explained as a recombinant between A and C strains, which co-occur in the region. Two further UK strains appeared to be recombinants between groups B and C. PMID- 12627487 TI - IL-10 gene polymorphism and herpesvirus infections. AB - Genetics has an important role in resistance to various infections and it also may modify the clinical picture of an infectious disease. Here, we briefly review our recent data demonstrating that the polymorphism of the IL-10 gene is associated with resistance to some common herpesviruses and, additionally, that this same gene is involved in the regulation of the severity of the infection and in the reactivation process. PMID- 12627489 TI - Varicella-zoster virus infection facilitates VZV glycoprotein E trafficking to the membrane surface of melanoma cells. AB - Varicella-zoster virus glycoprotein E (gE) is the most abundant VZV glycoprotein on the surface of virus-infected cells. VZV gE has targeting sequences for the trans-Golgi network (TGN) and is transported from the ER to the TGN in infected and gE-transfected cells. In this study, VZV gE expressing melanoma cell lines were generated. gE is expressed under the control of the reverse Tet repressor (Tet-On). gE induced by Tet-On is retained at the ER as well as in the cis Golgi by immunofluorescence confocal microscopy. To test whether other viral protein(s) may facilitate gE trafficking and surface localization, MSPgE-vOka virus that contains MSPgE in place of wt gE was made. MAb 3B3 anti-gE does not bind to MSPgE. This MAb was used to track the localization of gE in Met-gE cells post MSPgE-vOka infection. gE became detectable mostly at the TGN and on the cell surface after viral infection. These data indicate that viral proteins facilitate the trafficking and cell surface expression of gE. PMID- 12627488 TI - Analyses of the transcriptional pattern of glycoproteins E and I of Varicella zoster virus and evidence for a monocistronic transcription. AB - Glycoproteins I and E of the Varicella-zoster virus, encoded by the neighbouring open reading frames 67 and 68, are transcribed into several transcript species that differ in size. From gI, three transcripts of 1.65, 2.7, and 3.6 kb are known, and from gE, two transcripts of 2.15 and 3.6 kb in size are known. Here, we demonstrate that these various transcript species appear in different amounts at different times post infection. At 12 hr post infection, the transcripts of 1.65 (gI) and 2.15 (gE) were clearly detectable, whereas the other transcripts appeared later on. RT-PCR studies using a set of different primers provided clear evidence that gI and gE are transcribed both, mono- and bicistronically, with predominance on the respective monocistronic transcript. Additional evidence for monocistronic transcription was found in the fact that both glycoproteins contain their own transcriptional start sites. Both promoter regions have their own basal transcription activity and include active TATA-boxes that were recognized by the TATA-box binding protein. PMID- 12627490 TI - Infectibility of separated peripheral blood mononuclear cell subpopulations by varicella-zoster virus (VZV). AB - Varicella zoster-virus (VZV) is a humanpathogenic alpha-Herpesvirus that causes chickenpox after primary infection. The virus spreads by aerosol or direct contact with infectious vesical fluids, it enters the body via the respiratory tract. In a first viremic stage it replicates in local lymph nodes, followed by a secondary viremic stage. In the course it spreads through the body to endothelial cells in the periphery. During acute viremia of chickenpox viral DNA can be detected in peripheral blood mononuclear cells (PBMC) by PCR and in situ hybridization. Recently published results quantified the viral DNA load in PBMC and subpopulations by real-time PCR. In the animal SCID-hu mouse model system VZV showed a tropism for T-lymphocytes. The aim of this work was the investigation of viral ability to infect and to replicate in purified primary subtypes of PBMC, i.e., T-lymphocytes, B-lymphocytes, and monocytes. These cells were isolated from whole peripheral blood or tonsils and infected with cell-free VZV for different time periods. In all cell types, transcriptional activity was shown by amplification and detection of immediate early (IE) and late (L) viral mRNA by NASBA or RT-PCR. Expression of viral glycoproteins was analyzed and proved in lymphocytes by immunofluorescence microscopy. PMID- 12627491 TI - Identification of the authentic varicella-zoster virus gB (gene 31) initiating methionine overlapping the 3' end of gene 30. AB - The varicella-zoster virus (VZV) gB sequence was re-examined in light of recent knowledge about unusually long gB signal peptides in other herpesviral gB homologs. Through mutational analysis, the discovery was made that the authentic initiating methionine for VZV gB is a codon beginning at genome nucleotide 56,819. The total length for the VZV gB primary translation product was 931 amino acids (aa) with a 71-aa signal sequence. Considering the likely signal sequence cleavage site to be located between Ser 71 and Val 72, the length of the mature VZV gB polypeptide would then be 860 amino acids prior to further internal endoproteolytic cleavage between amino acids Arg 494 and Ser 495. In this report, we also produced a full-length gB and demonstrated its association with VZV gE, suggesting a possible gE-gB interaction during gB trafficking before its cleavage in the Golgi. PMID- 12627493 TI - Use of a rodent model to show that varicella-zoster virus ORF61 is dispensable for establishment of latency. AB - Varicella-zoster virus (VZV) results in a latent infection in humans after primary infection. Latency has also been established in guinea pigs and rats after inoculation with the virus. It was found that infection of cotton rats with the Oka vaccine strain of VZV results in a latent infection. To begin to identify which genes are required for latency, we infected cotton rats with VZV strain Oka that is deleted for ORF61. ORF61 protein transactivates certain VZV promoters and enhances the infectivity of viral DNA in transient transfections. Deletion of ORF61 results in abnormal syncytia and impairs the growth of VZV in vitro. Inoculation of cotton rats with ORF61-deleted Oka virus resulted in latent VZV infection in the nervous system similar to that seen for animals infected with parental virus. Thus, the cotton rat can be used to study the ability of mutants in the Oka vaccine strain of VZV to establish latent infection. PMID- 12627492 TI - Latent and lytic infection of isolated guinea pig enteric ganglia by varicella zoster virus. AB - Varicella zoster virus (VZV) has been demonstrated to infect guinea pig enteric neurons in vitro. Latent infection of isolated enteric neurons is established when the cultures predominantly consist of neurons and they are exposed to cell free VZV. Neurons harboring latent infection survive for weeks in vitro and express mRNA encoding ORFs 4, 21, 29, 40, 62, and 63, but not 14(gC) or 68 (gE) (although DNA encoding the glycoproteins is present). The expressed proteins are the same as those that are also expressed in human sensory neurons harboring latent VZV. In addition to mRNA, the immunoreactivities of ORFs 4, 21, 29, 62, and 63 can be detected. ORF 62 and 29 proteins are cytoplasmic and not intranuclear. VZV does not preferentially infect and/or become latent in intrinsic enteric primary afferent neurons indicating that the virus is latent in these neurons. Lytic infection occurs when mixed cultures of neurons and non neuronal cells of the bowel wall are exposed to cell-free VZV or when isolated enteric neurons are exposed to cell-associated VZV. When lytic infection occurs, enteric neurons die within 48 hr. Prior to their death, neurons express VZV glycoproteins, including gE and gB, and ORF 62 and 29 proteins are intranuclear. This new animal model should facilitate studies of VZV latency and the efficacy of therapies designed to prevent VZV infection, latency, and reactivation. PMID- 12627494 TI - Varicella-zoster virus isolates, but not the vaccine strain OKA, induce sensitivity to alpha-1 and beta-1 adrenergic stimulation of sensory neurones in culture. AB - The reactivation of varicella-zoster virus (VZV) from its persistent state in sensory neurones causes shingles and induces severe, long-lasting pain and hyperalgesia that often lead to postherpetic neuralgia. To investigate the VZV induced neuropathic changes, we established conditions for the active infection of sensory neurones from rat dorsal root ganglia in vitro. After 2 days of culture, up to 50% of the cells expressed viral antigens of the immediate-early and late replication phase. The intracellular calcium ion concentration was monitored in individual cells by microfluorimetry. Whereas the calcium response to capsaicin was preserved, the VZV-infected neurones gained an unusual sensitivity to noradrenaline stimulation in contrast to non-infected cells. The adrenergic agonists phenylephrine and isoproterenol had a similar efficacy demonstrating that both alpha(1)- and beta(1)-adrenoreceptors were involved. The sensitivity to adrenergic stimulation was observed after infection with different wildtype isolates, but not with the attenuated vaccine strain OKA. The lack of noradrenaline sensitivity of vaccine-infected neurones demands a structural comparison of wildtype and vaccine viruses with and without phenotype. A partial sequence evaluation (26 kb) of the European OKA vaccine strain surprisingly revealed a series of nucleotide exchanges in comparison to presumably identical OKA strains from other sources, although VZV is generally considered genetically stable. In summary, we report that the infection with wildtype VZV isolates, but not with the vaccine strain, induces noradrenaline sensitivity in sensory neurones, which correlates with clinical and experimental observations of adrenergic effects involved in VZV-induced neuralgia. PMID- 12627495 TI - Cellular factors and IE62 activation of VZV promoters. AB - The varicella zoster virus IE62 protein transactivates promoters representing all three kinetic classes of viral genes. This is accomplished in conjunction with the general transcription machinery of the cell and specific cellular transcription factors that act at sites located within viral promoters. The incidence of a subset of specific recognition sites was examined for the ubiquitous cellular factors Sp1 and USF within the VZV genome. Evidence is also presented for a direct physical interaction between Sp1 and IE62. PMID- 12627496 TI - Comparison of varicella-zoster virus ORF47 protein kinase and casein kinase II and their substrates. AB - ORF47, a serine/threonine protein kinase encoded by varicella-zoster virus (VZV), has often been compared to the ubiquitous cellular kinase, casein kinase II (CKII). However, no direct comparison of the two protein kinases has been carried out. Herein, we show that the ORF47 kinase was resistant to heparin, while CKII activity is profoundly inhibited by the acidic molecule in vitro. ORF47 required the presence of polyamines (aliphatic, positively-charged molecules) for in vitro activity. When polyamines were depleted from MeWo cells prior to VZV infection by pretreatment with D,L-alpha-difluoromethylornithine, VZV replication was reduced by 80%. Finally, the substrate specificity of the ORF47 kinase was defined using an in vitro assay. The ORF47 kinase phosphorylated maltose-binding protein, the mouse IgG2A heavy chain, the rabbit IgG heavy chain, casein, VZV ORF62, and VZV ORF63. The ORF47 kinase failed to phosphorylate an ORF62 truncation mutant, glutathione-S-transferase, or VZV gB. In contrast, CKII weakly phosphorylated VZV gB in vitro. By analyzing the sequences of these substrates, the minimal ORF47 consensus sequence was deduced to be the following motif: S/T-X-D/E-D/E, with a marked preference for additional acidic amino acids in the -1 and +1 position. PMID- 12627497 TI - Imaging of the varicella zoster virion in the viral highways: comparison with herpes simplex viruses 1 and 2, cytomegalovirus, pseudorabies virus, and human herpes viruses 6 and 7. AB - Imaging by scanning electron microscopy (SEM) can provide insight into viral egress. At a low magnification level, varicella zoster virions (VZV) emerge from an infected cell surface in a distinctive pattern previously described as "viral highways." Viral highways consist of thousands of particles arranged in linear pathways across the syncytial surface. This egress pattern has not been described with other herpesviruses, but a systematic analysis has not been performed. Therefore, the characteristic arrangement of VZV egress was compared with that of six other members of the herpes virus family, including herpes simplex virus (HSV) types 1 and 2, human cytomegalovirus (CMV), pseudorabies virus (PRV), and human herpesvirus types 6 and 7 (HHV-6 and HHV-7). Only VZV-infected cells exhibited viral highways. Subsequent SEM examination of VZ virions at an ultra high-resolution revealed that more than 70% were aberrant. Further imaging of the other herpesviruses demonstrated that VZV structure was more closely related to PRV than HSV-1 or HSV-2. Finally, it is noted that the individual members of the herpesvirus family have distinguishable SEM profiles. PMID- 12627498 TI - Varicella in Americans from NHANES III: implications for control through routine immunization. AB - At the time of varicella vaccine introduction in the United States, an estimated 4 million episodes of varicella occurred annually. This survey of varicella seroprevalence is the first to describe immunity to a vaccine-preventable disease prior to vaccine introduction in the United States population. The objective of this analysis is to describe patterns of naturally-acquired varicella and understand characteristics associated with infection in the varicella vaccine naive United States population. A nationally representative cross-sectional health examination survey that included venipuncture was conducted among 21,288 U.S. participants aged 6 years and older from 1988 through 1994. Serologic evidence of varicella-zoster virus infection was measured by enzyme immunoassay of varicella-zoster virus-specific IgG antibody. The seroprevalence of IgG antibody to varicella-zoster virus increased from 86.0% in children aged 6 through 11 years to 99.6% in adults aged 40 through 49 years. Immunity to varicella remained at 99% or higher in Americans aged 50 years and older. Among persons aged 6 through 19 years, non-Hispanic black children were 40% less likely to be seropositive compared with white children (odds ratio [OR], 0.6; 95% confidence interval [CI], 0.4-0.8). Among young adults aged 20 through 39 years, women with a history of live birth (OR, 4.3; 95% CI, 2.1-8.7) and married men (OR, 2.7; 95% CI, 1.2-5.7) were more likely to have naturally-acquired immunity to varicella. This study found that, prior to use of varicella vaccine in the United States, age, race, and marital characteristics were independently associated with naturally acquired varicella. Future varicella serosurveys in Americans will provide essential information to interpret the population impact of varicella vaccine. PMID- 12627499 TI - Factors influencing arm and axillary symptoms after treatment for node negative breast carcinoma. AB - BACKGROUND: The purpose of the current study was to identify the factors that contribute to postoperative arm symptoms following breast conserving surgery in a well-defined cohort of node negative breast carcinoma patients. METHODS: A convenience sample of 370 women >/= 50 years of age with node negative breast carcinoma who were participants in a randomized controlled trial designed to assess the need for breast radiation in addition to tamoxifen were surveyed. Axillary dissection was optional for patients 65 years or older and who were clinically node negative. RESULTS: A total of 65.1% (241/370) of women had ipsilateral shoulder or arm symptoms. Multivariate analysis revealed that axillary dissection, breast radiation, and younger age (odds ratio = 11.2, 1.65, and 3.8 respectively) were significantly (P < 0.05) associated with increased ipsilateral shoulder or arm symptoms. Treatment with axillary dissection and breast radiation were significant factors (P < 0.02) associated with the self reporting of arm swelling (odds ratio = 4.4 and 2.0, respectively). Patients 70 years old or greater reported significantly fewer arm symptoms (odds ratio = 0.26, P < 0.05) after axillary dissection. CONCLUSIONS: Arm symptoms were present in about 80% of patients who underwent breast conserving surgery, axillary dissection, and breast radiation in the current study. These symptoms were significantly associated with the use of axillary dissection, breast radiation, and younger age. Older patients experienced fewer arm symptoms after standard treatment for node negative breast carcinoma, and thus older age should not be a contraindication to axillary dissection. PMID- 12627500 TI - Flow cytometry and quantitative immunohistochemical study of cell cycle regulation proteins in invasive breast carcinoma: prognostic significance. AB - BACKGROUND: Between January 11, 1991 and January 8, 1992, 104 patients with previously untreated, invasive, primitive breast carcinoma were admitted to the authors' hospital. METHODS: For each patient, flow cytometry DNA analyses on frozen samples and on immunohistochemical staining were performed, including Ki 67, cyclin A, p53, and p21(waf1) (p21), with assessment of the percentages of positive nuclei were assessed. Correlations with classic clinicopathologic data and survival (overall, metastasis free, or recurrence free) and a multivariate analysis were performed. RESULTS: After a multivariate analysis according to a Cox model that was stratified by age, tumor size, tumor grade, lymph node status, and receptor status, among the factors studied, the presence of p21 was the unique remaining prognostic factor for patients with invasive breast carcinoma. Because of the lack of a correlation between p21 and proliferative factors (Ki 67, S-phase, and cyclin A), the authors combined p21 with those markers and found that, for the different combinations, after statistical analysis, only p21 combined with S-phase or with cyclin A and lymph node status were salient survival prognostic factors. CONCLUSIONS: Immunohistochemical study of proteins involved in the cell cycle and assessment of proliferative activity using flow cytometric DNA analysis aided the authors in singling out correlations of cyclin A and S-phase, along with p21, with metastasis free survival and overall survival in patients with invasive breast carcinoma. These promising results will require confirmation in a larger series of patients. PMID- 12627501 TI - Hormone replacement therapy containing progestins and given continuously increases breast carcinoma risk in Sweden. AB - BACKGROUND: The authors previously reported an increased risk of breast carcinoma with longer duration of hormone replacement therapy (HRT) use. It is unclear if different types of HRT confer different risks. METHODS: In this study, a population-based cohort of 29,508 women were interviewed during 1990-1992 to determine whether there are any differences in breast carcinoma risk according to different types and duration of HRT use. RESULTS: At the end of the follow-up period in December 2001, the cohort constituted 298,649 person-years. Slightly more breast carcinoma cases were seen (n = 556) than expected (n = 508.37; standardized morbidity ratio =1.09, 95% confidence interval [CI] = 1.00-1.19). Approximately 3663 women had ever used HRT. In Cox regression models, time to breast carcinoma in relation to duration and type of HRT use was analyzed, adjusting for age at menarche, age at first full-term pregnancy, parity, age at menopause, family history of breast carcinoma, and age at interview. In women with a natural menopause, a significantly higher risk was observed for longer duration of combined continuous HRT use compared with never users (hazard ratio [HR] = 4.60, 95% CI = 2.39-8.84). Nonsignificant elevated risks also were observed for longer combined sequential (HR = 2.23, 95% CI = 0.90-5.56), gestagen only (HR = 3.74,9 5% CI = 0.94-14.97), and estriol use (HR = 1.89, 95% CI = 0.81 4.39). No increased risk was seen in women after 5 years of nonuse. When studying women who ever used only one type of HRT, even more elevated HRs for gestagen containing preparations were seen. The highest risks were associated with the combined continuous and gestagen-only therapy in women with >/= 48 months of use. Use of estradiol without progestins did not increase breast carcinoma risk significantly. The authors estimated the cumulative risk of breast carcinoma in a 50-year-old woman with gestagen-containing therapies for >/= 48 months, with a follow-up of 10 years, to be 7% (95% CI = 5.4-11.4%) compared with 2% (95% CI = 1.6%-2.9%) for never-users of HRT. CONCLUSIONS: Longer use of HRT containing progestins significantly elevates breast carcinoma risk whereas estradiol use does not. Continued use of progestins rendered the highest risks. The yearly risk of breast carcinoma for long-term users of progestins is of the magnitude of 50% the risk of a BRCA1 mutation carrier. PMID- 12627503 TI - Lung carcinoma after radiation therapy in women treated with lumpectomy or mastectomy for primary breast carcinoma. AB - BACKGROUND: Prior studies have demonstrated that women who receive adjuvant radiation therapy (RT) after mastectomy for breast carcinoma have an increased risk of a second primary lung carcinoma after 10 years, but, to the authors' knowledge, the risk associated with adjuvant RT after breast-conserving surgery (lumpectomy) has yet to be determined. The purpose of the current study was to confirm and extend earlier findings of the effects of postmastectomy RT on second primary lung carcinoma and to investigate the impact of postlumpectomy RT on second primary lung carcinoma in the same population and to compare the results. METHODS: The authors used data from the population-based Surveillance, Epidemiology, and End Results (SEER) program of the U.S. National Cancer Institute, encompassing approximately 10% of the U.S. population, from 1973 to 1998. Of the women with nonmetastatic invasive breast carcinoma, 194,981 had been treated with mastectomy and 65,560 were treated with lumpectomy. Cox regression analysis was used to estimate the relative risk (RR) of a second primary lung carcinoma among women treated with RT compared with those who received surgery alone as a function of time interval since treatment and laterality for both types of surgery. RESULTS: Although no statistically significant elevation in risk for second primary lung carcinoma prior to 10 years was observed, the authors estimated a RR of 2.06 (95% confidence interval [95% CI], 1.53-2.78) and 2.09 (95% CI, 1.50-2.90) for ipsilateral lung carcinoma at 10-14 years and 15+ years after postmastectomy RT, respectively, whereas no increased risk was observed for the contralateral lung. The excess risk of ipsilateral lung carcinoma after postmastectomy RT was found for all three major histologic subtypes of lung carcinoma (adenocarcinoma, squamous cell carcinoma, and small cell carcinoma). No increased risk of lung carcinoma was observed at 10-14 years after postlumpectomy RT for either lung. CONCLUSIONS: Postmastectomy RT was found to provide a moderate increase in risk for ipsilateral lung carcinoma starting 10 years after exposure; this increased risk is reported to persist to at least 20 years. Postlumpectomy RT does not appear to incur an increased risk. These findings should be reassuring to women treated with either type of RT, but the excess risk in the postmastectomy group should be considered in the choice between treatment options. PMID- 12627502 TI - Differences in the pathologic and molecular features of intraductal breast carcinoma between younger and older women. AB - BACKGROUND: Patients diagnosed with ductal carcinoma in situ (DCIS) at a young age appear to have a different natural history and biology, including a higher local relapse rate, than patients diagnosed later in life. The current study compared various pathologic and molecular features of DCIS arising in a cohort of young women with those of DCIS arising in a cohort of older women to identify potential biologic differences between these two populations of patients. METHODS: The study population consisted of 20 patients age < 42 years and 34 patients age > 60 years who were treated at Yale University School of Medicine with breast-conserving therapy (BCT) and whose archival paraffin blocks were available and had sufficient tumor for staining. The original slides from each case were reviewed and the most representative specimen block from each case was processed for immunohistochemical staining. Pathologic characteristics evaluated for each case included histology, grade, and presence of necrosis. Paraffin embedded sections were immunohistochemically evaluated for expression of HER 2/neu, estrogen receptor (ER), progesterone receptor (PR), bcl-2, cyclin D1, Ki 67, and p53. RESULTS: Although there was no difference in pathologic features of the tumors between the two groups, HER-2/neu was found to be overexpressed in a greater percentage of the younger population (P = 0.06). There was no apparent difference in expression of the other markers. Of note, HER-2/neu expression was correlated with high nuclear grade (P = 0.004), necrosis (P = 0.06), and ER and PR negativity (P = 0.01 and P = 0.03, respectively) in the combined population. CONCLUSIONS: The current study data suggested that HER-2/neu overexpression in younger patients may characterize a biologic difference in their tumor and may partially contribute to their higher risk of recurrence. Further studies are needed to assess whether this difference holds independent of grade and to evaluate the prognostic significance of HER-2/neu overexpression in DCIS. PMID- 12627504 TI - Combination of tumor necrosis factor-alpha with sulindac augments its apoptotic potential and suppresses tumor growth of human carcinoma cells in nude mice. AB - BACKGROUND: Resistance to apoptosis may be responsible for a principal mechanism by which cancer cells overcome anticancer therapies. Among antiapoptotic signals, the transcription factor, NF-kappaB, plays a pivotal role in the resistance because it is frequently activated in many primary carcinoma cells. However, NF kappaB is also activated by several anticancer therapies, including tumor necrosis factor-alpha (TNF-alpha). The NF-kappaB-mediated survival signals are supposed to evade these therapies. Recently, a nonsteroidal antiinflammatory drug, sulindac, and its metabolites have been shown to inhibit the NF-kappaB pathway and to enhance TNF-alpha-mediated apoptosis in lung carcinoma cell lines. In the current study, the authors investigated whether sulindac can augment TNF alpha-mediated apoptosis in other human carcinoma cells and whether it can be applied for in vivo clinical usage. METHODS: Human gastric MKN45 and cervical HeLa carcinoma cells were treated with sulindac and/or TNF-alpha. Proapoptotic effects of these agents were evaluated by trypan blue exclusion assay, DNA fragmentation, and caspase-3 activity. The effect of sulindac on NF-kappaB activation was evaluated by luciferase reporter and gel-shift assays. The suppressive effects of these reagents on the subcutaneous tumor growth of MKN45 cells were evaluated by measuring tumor size in nude mice. RESULTS: Sulindac inhibited TNF-alpha-mediated NF-kappaB activation and greatly sensitized MKN45 and HeLa cell lines to TNF-alpha. Moreover, in vivo tumor growth of MKN45 cells was inhibited most strongly by a combination of TNF-alpha with sulindac compared with TNF-alpha or sulindac alone. CONCLUSIONS: The current study data strongly suggest that combination therapy of TNF-alpha with sulindac may sensitize tumor cells to TNF-alpha and augment its proapoptotic potential. Therefore, in combination with sulindac, TNF-alpha may become a potentially useful anticancer agent to suppress tumor growth in a wide range of carcinomas. PMID- 12627505 TI - Frequency of loss of hMLH1 expression in colorectal carcinoma increases with advancing age. AB - BACKGROUND: The correlation between age at diagnosis and loss of expression of hMLH1 protein in patients with colorectal carcinoma (CRC) has not been evaluated systematically. METHODS: Immunohistochemistry was performed for hMLH1 protein in tumor samples from 867 patients with CRC. The authors defined tumors arising in the cecum, ascending colon, and transverse colon as right-sided and tumors arising in the descending colon, sigmoid colon, and rectum as left-sided. Patients' gender, tumor location (side), and hMLH1 expression were analyzed by age groups. RESULTS: The percentage of tumors with hMLH1 expression loss increased significantly with advancing age (P < 0.0001): There were no tumors in patients age < 40 years that manifested loss of hMLH1 expression, compared with 29% of tumors that manifested loss of hMLH1 expression in patients age > 90 years. Loss of hMLH1 expression occurred more often in patients with right-sided tumors (32.7% vs. 5.2% of patients with left-sided tumors; P < 0.0001) and in tumors from female patients (24.3% vs. 11.5% of tumors from male patients; P < 0.0001). There was no evidence of interaction between gender and tumor location. CONCLUSIONS: Loss of hMLH1 expression in patients with CRC was associated strongly with increasing age. hMLH1 expression loss was more pronounced in tumors from female patients and in tumors that originated on the right side of the colon. Loss of hMLH1 expression in right-sided tumors occurred in nearly 50% of patients age > 90 years. This age-related trend also was observed for males and in tumors that originated in the left colon. PMID- 12627507 TI - Therapy and outcome of small cell carcinoma of the kidney: report of two cases and a systematic review of the literature. AB - BACKGROUND: Primary small cell carcinoma originating from the kidneys is an extremely rare neoplasm. The authors described two patients with small cell carcinoma of the kidney and provided a systematic review of the literature to detail the clinical characteristics and therapy of this rare tumor. METHODS: MEDLINE and CANCERLIT literature search was performed from 1966 to 2002 for articles on small cell carcinoma of the kidney. Twenty-two patients with small cell carcinoma of the kidney and renal pelvis were reviewed. RESULTS: The median age at diagnosis was 62 years and there was a female preponderance (male:female ratio, 1:3.4). Abdominal pain (70%) was the most commonly reported symptom. Distant metastases were present in 32% of patients at the time of diagnosis. Surgery and systemic chemotherapy were the primary therapeutic modalities utilized (nephrectomy alone: 9 patients; nephrectomy and chemotherapy: 10 patients; chemotherapy alone: 3 patients). The median survival was 8 months (range, < 1-101 months). The use of platinum-based chemotherapy was predictive of an improved overall survival (median survival was 20 months in patients receiving a platinum-containing regimen compared with 8 months in those who did not receive platinum; P = 0.02). CONCLUSIONS: Small cell carcinoma of the kidney is an extremely rare neoplasm that resembles its counterparts arising from the tracheo bronchial and other extrapulmonary sites in its aggressive behavior and high propensity for locoregional and distant dissemination. Clinical presentation is usually late in the course of the disease. The use of platinum-based chemotherapy has been associated with tumor regression and prolonged survival. PMID- 12627506 TI - Use of quality indicators to evaluate the care of patients with localized prostate carcinoma. AB - BACKGROUND: The goal of quality assurance in health care is to preserve and improve patient care. Recently, RAND developed a set of evidence-based candidate indicators for evaluating the quality of care for patients with localized prostate carcinoma; however, the feasibility and sensitivity of these indicators have not been tested in a clinical setting. The objectives of this study were to evaluate the feasibility of measuring these quality indicators and to determine their sensitivity to change in practice patterns over time. METHODS: One hundred sixty-eight men who presented in either 1995 or in 2000 and were treated for localized prostate carcinoma were selected randomly from the University of Michigan tumor registry. A combination of electronic data base review and explicit chart review was used to assess the feasibility of measuring compliance for each indicator. For each indicator in which assessment was feasible, compliance with the RAND indicators was determined for patients in both years. Multivariate regression analysis was used to adjust for potential confounding effects of disease stage, tumor grade, prostate specific antigen (PSA) level, patient age, and therapy. RESULTS: Based on review of available clinical data, measurement of compliance was feasible for 19 of 22 RAND candidate quality indicators (86%). For five indicators, significant differences in documentation (compliance) were detected between 1995 and 2000 (P < 0.05). Treatment received and higher PSA levels were associated independently with documentation of compliance for several indicators (P < 0.05). CONCLUSIONS: Measurement of the majority of the RAND quality indicators for the treatment of patients with localized prostate carcinoma was feasible, and improvements in several indicators were observed between 1995 and 2000. Demonstration of such variation, even within a single institution, suggests that the indicators are sufficiently sensitive to detect differences in practice patterns. PMID- 12627509 TI - Down-regulation of fragile histidine triad expression in prostate carcinoma. AB - BACKGROUND: The fragile histidine triad (FHIT) gene is a tumor suppressor gene that belongs to the histidine triad family of nucleoside binding proteins. The gene encompasses the common human chromosomal fragile site, the FRA3B locus at chromosome 3p14.2, and is expressed in most normal adult tissues and tumor cell lines. Numerous studies have indicated that the FHIT gene on chromosome 3p may play an important role in human neoplasia, although very few studies have investigated the FHIT gene in prostate carcinoma. METHODS: Using immunohistochemical analyses, the authors studied the expression of FHIT in prostate tumors from 84 radical prostatectomy specimens to determine whether there were any correlations between FHIT expression and various clinicopathologic characteristics. RESULTS: The percentages of cells stained with antibody to FHIT were significantly lower overall for tumor cells compared with normal cells (P = 0.0001). FHIT immunostaining intensity also was significantly lower for tumor cells compared with normal cells (P = 0.0001). A weak but statistically significant correlation (P = 0.045) was demonstrated with the presence of extraprostatic extension in the patient samples. No other significant correlation was seen between the percentage of cells stained for FHIT or FHIT immunostaining intensity and Gleason grade, tumor stage, tumor size, lymph node metastasis, surgical margins, vascular invasion, perineural invasion, or the presence of high grade prostatic intraepithelial neoplasia. CONCLUSIONS: The data presented indicate a down-regulation of the FHIT tumor suppressor gene in prostate carcinoma and, thus, propose a potential target for therapeutic intervention. PMID- 12627510 TI - Prognostic index for patients with parotid carcinoma: external validation using the nationwide 1985-1994 Dutch Head and Neck Oncology Cooperative Group database. AB - BACKGROUND: Validation of the prognostic indices for the recurrence-free interval of patients with parotid carcinoma, the development of which was described in a previous report, is needed to be confident of their generalizability and justified prospective use. METHODS: The Dutch Cooperative Group on Head and Neck Cancer database contains 231 patients with parotid carcinoma from six tertiary referral centers who were seen between 1985-1994. This database was used to validate the predictive value of the prognostic indices, PS1 (pretreatment index predicting recurrence) and PS2 (posttreatment index predicting recurrence), in patients with parotid carcinoma. Validation methods included calculation of both indices for each patient, comparison of coefficients, construction of survival curves using the published cutoff points, and calculation of concordance measure C. Wald tests for optimization of scale and weights of the contributing variables, and for possible score improvement by including other variables, were also performed. RESULTS: The 5-year disease-free rate was 62% (standard error = 4%). PS1 was calculated for 183 patients. The previously set cutoff points resulted in 5-year disease-free rates that ranged from 92% (PS1 = 1) to 42% for the least favorable group (PS1 = 4). Concordance measure C was 0.74. The postoperative score, PS2, was calculated for 171 patients. Previous cutoff points resulted in 5-year disease-free rates that ranged from 90% (PS2 = 1) to 40% (PS2 = 4). Concordance measure C was 0.71. Both PS1 and PS2 did not improve using the findings in this independent material. CONCLUSIONS: The prognostic indices performed adequately in this validation sample. This finding demonstrates that the prognosis of a parotid carcinoma patient can be quantified by using a weighted combination of the parameters of age, pain, clinical T (cT) classification, clinical N (cN) classification, skin invasion, facial nerve dysfunction, perineural growth, and involved surgical margins. A quantified prognosis, in terms of the recurrence-free interval, can be used to provide information to the patient, to perform clinical trials, and eventually to make clinical decisions. A user-friendly translation and computerized calculation of the indices should be the next step toward generalized prospective use and repetitive evaluation of the indices. PMID- 12627508 TI - A phase II trial of green tea in the treatment of patients with androgen independent metastatic prostate carcinoma. AB - BACKGROUND: Recent laboratory and epidemiologic studies have suggested that green tea has antitumor effects in patients with prostate carcinoma. This Phase II trial explored green tea's antineoplastic effects in patients with androgen independent prostate carcinoma. METHODS: This study, which was conducted by the North Central Cancer Treatment Group, evaluated 42 patients who were asymptomatic and had manifested, progressive prostate specific antigen (PSA) elevation with hormone therapy. Continued use of luteinizing hormone-releasing hormone agonist was permitted; however, patients were ineligible if they had received other treatments for their disease in the preceding 4 weeks or if they had received a long-acting antiandrogen therapy in the preceding 6 weeks. Patients were instructed to take 6 grams of green tea per day orally in 6 divided doses. Each dose contained 100 calories and 46 mg of caffeine. Patients were monitored monthly for response and toxicity. RESULTS: Tumor response, defined as a decline >/= 50% in the baseline PSA value, occurred in a single patient, or 2% of the cohort (95% confidence interval, 1-14%). This one response was not sustained beyond 2 months. At the end of the first month, the median change in the PSA value from baseline for the cohort increased by 43%. Green tea toxicity, usually Grade 1 or 2, occurred in 69% of patients and included nausea, emesis, insomnia, fatigue, diarrhea, abdominal pain, and confusion. However, six episodes of Grade 3 toxicity and one episode of Grade 4 toxicity also occurred, with the latter manifesting as severe confusion. CONCLUSIONS: Green tea carries limited antineoplastic activity, as defined by a decline in PSA levels, among patients with androgen independent prostate carcinoma. PMID- 12627511 TI - Extent of extracapsular spread: a critical prognosticator in oral tongue cancer. AB - BACKGROUND: Extracapsular spread (ECS) of metastatic squamous cell carcinoma of the head and neck to regional lymph nodes is the most reliable predictor of poor treatment outcomes. Recently, the authors have shown that ECS is significantly associated with higher rates of locoregional recurrence, distant metastasis, and decreased survival in patients with squamous cell carcinoma of the oral tongue (SCCOT). The purpose of this review was to determine if the degree of ECS impacts distant metastasis rates and survival. METHODS: Two hundred sixty-six patients treated for SCCOT with surgery +/- adjuvant radiotherapy from 1980-1995 were reviewed. The setting was a tertiary referral center. The extent of ECS on histopathologic review of involved lymph nodes was measured from the capsular margin to the farthest perinodal extension in mm. Extent of ECS and the number of pathologic lymph nodes with or without ECS were analyzed for disease-free interval, survival rates, and distant metastases. RESULTS: No differences in the survival of patients with ECS of 2 mm was found (P = 0.92). Patients with both ECS and multiple positive lymph nodes had decreased overall survival (P = 0.0003), disease-specific survival (P = 0.0005), and a shorter disease-free interval (P = 0.019) when compared with those with a single positive lymph node with ECS. Those with multiple ECS+ lymph nodes had the worst prognosis (P = 0.001). CONCLUSIONS: Based on these findings, the authors recommended that all patients with SCCOT with ECS or multiple positive lymph nodes with or without ECS on pathologic review be considered for clinical trials that intensify regional and systemic adjuvant therapy. PMID- 12627512 TI - Lineage specific treatment of adult patients with acute lymphoblastic leukemia in first remission with anti-B4-blocked ricin or high-dose cytarabine: Cancer and Leukemia Group B Study 9311. AB - BACKGROUND: Anti-B4-blocked ricin is an immunotoxin comprised of an anti-CD19 murine monoclonal antibody (B4) conjugated to blocked ricin, which has cytotoxic activity in patients with lymphoid malignancies. METHODS: Adults with untreated acute lymphoblastic leukemia (ALL) were treated with a previously developed and tested chemotherapeutic regimen. Patients with CD19 positive ALL were given anti B4-blocked ricin as 2 7-day continuous infusions 1 week apart. Patients with CD19 negative ALL received high-dose cytarabine. Serial polymerase chain reaction (PCR) assays of BCR-ABL, immunoglobulin heavy chain (IGH), and T-cell receptor (TCR) genes were used to measure the impact of lineage specific intensification treatment on minimal residual disease. RESULTS: Eighty-two adults were enrolled, and 78 were eligible. The median age was 34 years (range, 17-81 years). Sixty-six patients (85%) achieved complete remission. Forty-six patients received the anti B4-blocked ricin, which generally was well tolerated; 80% were able to receive both courses. The most common toxicity was asymptomatic transient elevation of liver function tests in 72% of patients. Lymphopenia occurred in 46% of patients. Two patients developed antibodies to the anti-B4-blocked ricin. Molecular monitoring before and after the experimental course of intensification did not show a consistent change in the number of leukemia cells remaining, and the immediate posttreatment PCR studies did not correlate with remission duration. CONCLUSIONS: Intensification therapy with anti-B4-blocked ricin is feasible for patients with CD19 positive ALL, although there is little evidence of an additional clinical benefit from the anti-B4-blocked ricin. Cancer 2003;97:1471 80. PMID- 12627514 TI - Tumor mitotic rate is a more powerful prognostic indicator than ulceration in patients with primary cutaneous melanoma: an analysis of 3661 patients from a single center. AB - BACKGROUND: The current study was performed to determine whether tumor mitotic rate (TMR) is a useful, independent prognostic factor in patients with localized cutaneous melanoma. METHODS: From the Sydney Melanoma Unit database, 3661 patients with complete clinical information and details of primary tumor thickness, ulcerative state, and TMR were studied. TMR was expressed as mitoses per mm(2) in the dermal part of the tumor in which most mitoses were seen, as recommended in the 1982 revision of the 1972 Sydney classification of malignant melanoma. To determine which was the more prognostically useful method of grouping TMR, two separate methods (A and B) were used. Factors predicting melanoma-specific survival were analyzed using the Cox proportional hazards regression model. RESULTS: Patients with a TMR of 0 mitoses/mm(2) had a significantly better survival than those with 1 mitosis/mm(2) (P < 0.0001) but no significant survival differences were recorded for the stepwise increases from 1 2, 2-3, 3-4, and 4-5/mm(2). Tumor thickness, ulceration, and TMR were closely correlated, whether TMR was grouped using Method A (0, 1-4, 5-10, and >/= 11 mitoses/mm(2)) or Method B (0-1, 2-4, and >/= 5 mitoses/mm(2)). However, Cox regression analysis indicated that the TMR was a highly significant independent prognostic factor, particularly when grouped according to Method A, in which it was second only to tumor thickness as the most powerful predictor of survival (P < 0.0001). CONCLUSIONS: TMR is an important independent predictor of survival for melanoma patients. If confirmed by studies from other centers, it has the potential to further improve the accuracy of melanoma staging, as well as to define more rigidly the risk categories for patients entering clinical trials. PMID- 12627515 TI - Barriers to the participation of African-American patients with cancer in clinical trials: a pilot study. AB - BACKGROUND: African-American patients have been under-represented in oncology clinical trials. Better understanding barriers to African-American participation may help increase the accrual of African-American patients onto clinical trials. METHODS: Two hundred eighteen patients with malignant disease (72 African American patients and 146 white patients) were recruited from the Duke Cancer Clinic and from Duke Oncology Outreach Clinics (DOORS). Patients were interviewed using a standardized survey. Questions included patients' knowledge of cancer, religious/spiritual beliefs, satisfaction with medical care, knowledge of clinical trials, reasons for participating or refusing to participate in a clinical trial, financial/transportation issues, and demographic factors, such as age and education. Data on attitudes and belief were analyzed for group differences between African-American patients and white patients as well as between patients who were treated at the Duke Cancer Clinic and patients who were treated at DOORS clinics. RESULTS: Willingness to participate in a clinical trial depended on both race and clinic site. Forty-five percent of white patients, compared with 31% of African-American patients, were willing to participate in a clinical trial (P = 0.05). white and African-American patients who were treated at the Duke Cancer Clinic were more willing to participate in a trial compared with their counterparts who were treated at DOORS clinics (47% vs. 37%, respectively; P = 0.09). The greatest differences between groups (African American patients vs. white patients and Duke Cancer Clinic patients vs. DOORS patients) were education and income: Much greater percentages of African-American patients and DOORS patients did not complete high school and had annual incomes < $15,000. In addition, more African-American patients than white patients believed that God would determine whether they would be cured or would die from their disease. In a multivariate analysis, education, income, and belief that God would determine the patient's outcome also were correlated with a decreased willingness to participate in clinical trials. CONCLUSIONS: Factors associated with religion, education, and income, rather than race, may be major barriers to clinical trial participation. Interventions that target education and income may increase the recruitment of African-American oncology patients onto clinical trials. PMID- 12627513 TI - Gemtuzumab, fludarabine, cytarabine, and cyclosporine in patients with newly diagnosed acute myelogenous leukemia or high-risk myelodysplastic syndromes. AB - BACKGROUND: Gemtuzumab is used to treat patients with previously untreated or recurrent acute myelogenous leukemia (AML). The fludarabine and cytarabine (ara C) regimen is active in these patients. Resistance to gemtuzumab is associated with blast multidrug resistance (MDR). The objectives of this study were to evaluate the efficacy and toxicity of a combination regimen of gemtuzumab, fludarabine, ara-C, and the MDR modifier (cyclosporine [CyA]) in patients with previously untreated AML, refractory anemia with excess blasts (RAEB), or RAEB in transformation (RAEBT). METHODS: The MFAC regimen was comprised of gemtuzumab (Mylotarg trade mark ) (6 mg/m(2) intravenously [i.v.] on Day 1); fludarabine and ara-C (15 mg/m(2) and 0.5 g/m(2), respectively, twice daily on Days 2-6); and CSA (6 mg/kg loading dose before gemtuzumab, followed by 16 mg/kg continuous i.v. infusion on Days 1 and 2). RESULTS: Fifty-nine evaluable patients were treated: 39 patients (66%) had AML and 20 patients (34%) had RAEB/RAEBT. Their median age was 57 years (range, 27-76 years). The MFAC regimen induced complete remission (CR) in 27 patients (46%) and CR with incomplete platelet recovery (CRp) in 1 patient (2%). The median survival period is 8 months. At 12 months, the survival rate is 38% and the event-free survival rate in patients with CR/CRp is 27%. Infections complicated 38% of the courses of chemotherapy. Grade 3/4 toxicity included hyperbilirubinemia in 31% and transaminitis in 7% of the patients. Four patients (7%) developed hepatic venoocclusive disease (VOD). CONCLUSIONS: The MFAC regimen may merit further study in patients with AML if measures to avoid and/or treat VOD can be incorporated into the regimen. PMID- 12627516 TI - Trends in prostate cancer mortality among black men and white men in the United States. AB - BACKGROUND: Prostate cancer mortality rates in the United States declined sharply after 1991 in white men and declined after 1992 in black men. The current study was conducted to investigate possible mechanisms for the declining prostate cancer mortality rates in the United States. METHODS: The authors examined and compared patterns of prostate cancer incidence, survival rates, and mortality rates among black men and white men in the United States using the 1969-1999 U.S. prostate cancer mortality rates and the 1975-1999 prostate cancer incidence, survival, and incidence-based mortality rates from the Surveillance, Epidemiology, and End Results (SEER) Program for the U.S. population. The SEER data represent approximately 10% of the U.S. population. RESULTS: Prostate cancer incidence and mortality rates showed transient increases after 1986, when the U.S. Food and Drug Administration approved the use of prostate specific antigen (PSA) testing. The age-adjusted prostate cancer mortality rates for men age 50-84 years, however, have dropped below the rate in 1986 since 1995 for white men and since 1997 for black men. In fact, for white men ages 50-79 years, the 1998 and 1999 rates were the lowest observed since 1950. Incidence-based mortality rates by disease stage revealed that the recent declines were due to declines in distant disease mortality. Moreover, the decrease in distant disease mortality was due to a decline in distant disease incidence, and not to improved survival of patients with distant disease. CONCLUSIONS: Similar incidence, survival, and mortality rate patterns are seen in black men and white men in the United States, although with differences in the timing and magnitude of recent rate decreases. Increased detection of prostate cancer before it becomes metastatic, possibly reflecting increased use of PSA testing after 1986, may explain much of the recent mortality decrease in both white men and black men. PMID- 12627517 TI - Health care disparities in older patients with breast carcinoma: informational support from physicians. AB - BACKGROUND: Little is known about how disparities in the treatment of patients with breast carcinoma based on patient age and ethnicity are effected or mitigated at the patient-physician interaction level. The objectives of this study were to document physician provision of informational support to patients at the time of a new diagnosis of breast carcinoma and to assess differences according to patient age and ethnic group in terms of the information received and desired. METHODS: Participants were 222 patients with breast carcinoma in Los Angeles County, California, age >/= 55 years who were interviewed within 6 months of their diagnosis of breast carcinoma and/or within 1 month posttreatment. Respondents were asked about receipt and helpfulness of 10 tangible informational support items (e.g., whether booklets, videotapes, medical records, etc. were provided by physicians) and 15 interactive informational support items (e.g., whether physicians discussed breast cancer topics, such as risk of recurrence or treatment options). An index of the tangible informational support items and a scale of the interactive informational support items received were created for summary analyses. Patients' medical records were abstracted for breast carcinoma stage and treatment type; surgeons also were surveyed about sociodemographic and practice characteristics. RESULTS: In multiple linear regression analyses, older age (beta coefficient [beta] +/- standard error [SE], - 0.08 +/- 0.02; P = 0.001) and Latina ethnicity (beta +/- SE, - 1.21 +/- 0.40; P = 0.003) had a negative association with physician provision of interactive informational support, controlling for patient and physician sociodemographic characteristics, practice characteristics, breast carcinoma stage, comorbidity, number of physicians seen, visit length, social support, and patient self-efficacy in interacting with physicians (adjusted correlation coefficient [R(2)] for the model, 0.33; P < 0.00001). Both older patients and ethnic minority patients, as well as their respective comparison groups, rated most breast cancer information as at least as helpful. Both groups preferred interpersonal sources of information to written sources, although they received interpersonal sources less frequently. CONCLUSIONS: Older patients and Latina patients with breast carcinoma received less interactive informational support from their physicians compared with younger patients, differences that persisted after controlling for a wide range of sociodemographic, psychosocial, and physician factors. Improving the quality of communication at the patient-physician interaction level may be an important avenue to reducing age and ethnic group treatment disparities among patients with breast carcinoma. PMID- 12627518 TI - Progress in cancer screening practices in the United States: results from the 2000 National Health Interview Survey. AB - BACKGROUND: Understanding differences in cancer screening among population groups in 2000 and successes or failures in reducing disparities over time among groups is important for planning a public health strategy to reduce or eliminate health disparities, a major goal of Healthy People 2010 national cancer screening objectives. In 2000, the new cancer control module added to the National Health Interview Survey (NHIS) collected more detailed information on cancer screening compared with previous surveys. METHODS: Data from the 2000 NHIS and earlier surveys were analyzed to discern patterns and trends in cancer screening practices, including Pap tests, mammography, prostate specific antigen (PSA) screening, and colorectal screening. The data are reported for population subgroups that were defined by a number of demographic and socioeconomic characteristics. RESULTS: Women who were least likely to have had a mammogram within the last 2 years were those with no usual source of health care (61%), women with no health insurance (67%), and women who immigrated to the United States within the last 10 years (61%). Results for Pap tests within the last 3 years were similar. Among both men and women, those least likely to have had a fecal occult blood test or endoscopy within the recommended screening interval had no usual source of care (14% for men and 18% for women), no health insurance (20% for men and 18% for women), or were recent immigrants (20% for men and 18% for women). An analysis of changes in test use since the 1987 survey indicates that the disparities are widening among groups with no usual source of care. CONCLUSIONS: No striking improvements have been observed for the groups with greatest need. Although screening use for most groups has increased since 1987, major disparities remain. Some groups, notably individuals with no usual source of care and the uninsured are falling further behind; and, according to the 2000 data, recent immigrants also experience a significant gap in screening utilization. More attention is needed to overcome screening barriers for these groups if the population benefits of cancer screening are to be achieved. PMID- 12627519 TI - Cost of chemotherapy-induced thrombocytopenia among patients with lymphoma or solid tumors. AB - BACKGROUND: The purpose of this study was to estimate the mean incremental cost of chemotherapy-induced thrombocytopenia and the drivers of cost. Another goal was to estimate the impact of depth and duration of thrombocytopenia on the cost of thrombocytopenia. METHODS: A retrospective cohort, consisting of a random sample of 75 solid tumor or lymphoma patients who developed chemotherapy-induced thrombocytopenia (/= 12.0 and therefore could be characterized as suffering from fatigue. This group of patients had a significantly lower QOL, had higher scores of anxiety and depression, and perceived that they had less social support. In a multiple regression model, mental adjustment, social support, anxiety, and depression as well as fatigue were significant predictors of QOL (FACT-generic part total score) whereas clinical and sociodemographic variables were not. CONCLUSION: A remarkably high proportion of ovarian carcinoma survivors suffered from fatigue. Because this symptom is a key predictor of QOL, it should be given more attention in aftercare programs. PMID- 12627523 TI - Levels of hypoxia-inducible factor-1alpha independently predict prognosis in patients with lymph node negative breast carcinoma. AB - BACKGROUND: Hypoxia-inducible factor-1 (HIF-1) is a transcription factor that plays an important role in tumor growth and metastasis by regulating energy metabolism and inducing angiogenesis to survive cellular hypoxia. Increased levels of HIF-1alpha, the O(2)-regulated subunit of HIF-1, were noted during breast carcinogenesis. In this study, the prognostic value of HIF-1alpha expression and its correlation with various clinicopathologic variables in patients with invasive breast carcinoma were investigated. METHODS: Expression levels of HIF-1alpha, HER-2/neu, estrogen receptor, and progesterone receptor were analyzed in 150 patients with early-stage breast carcinoma by immunohistochemistry. HER-2/neu gene amplification was investigated with automated fluorescent in situ hybridization. The mitotic activity index, histologic grade, and tumor type were assessed in hematoxylin and eosinstained specimens. Clinical data included disease-free survival, overall survival, lymph node status, and tumor size. The data were analyzed with two-sided univariate and multivariate tests, with P values < 0.05 considered significant. RESULTS: High levels of HIF-1alpha had an association of borderline significance with decreased overall survival (P = 0.059) and disease-free survival (P = 0.110) that was ascribed completely to the subgroup of women with lymph node negative tumors (n = 81 patients; P = 0.008 and P = 0.004, respectively). HER-2/neu immunoreactivity (P < 0.001) and gene amplification (P < 0.001), vascular endothelial growth factor expression (P = 0.016), and Ki-67 expression (P < 0.001) were correlated strongly with HIF-1alpha positivity, although none of those factors had an independent effect on survival. CONCLUSIONS: Increased levels of HIF-1alpha were associated independently with shortened survival in patients with lymph node negative breast carcinoma. Therefore, the use of immunohistochemical assessment of HIF-1alpha as a new predictor of poor outcome may improve clinical decision making regarding adjuvant treatment of patients with lymph node negative breast carcinoma. PMID- 12627524 TI - Low molecular weight inhibitors of matrix metalloproteinases can enhance the expression of matrix metalloproteinase-2 (gelatinase A) without inhibiting its activation. AB - BACKGROUND: In the current study, the authors investigated the effects of synthetic low molecular weight inhibitors of matrix metalloproteinases (MMPs) on the expression and activation of MMP-2 in a three-dimensional tissue system. METHODS: Rabbit periosteal explants were cultured with or without various concentrations of the MMP inhibitors CT1166, CT1399, or CT1746, and conditioned media and tissue extracts were analyzed for the expression and activity of MMP-2. RESULTS: The data showed that blocking the activity of all MMPs with relatively high inhibitor concentrations completely prevented the conversion of pro-MMP-2 into its active form and that the level of protein was decreased. Selective inhibition of the activity of gelatinases (MMP-2 and MMP-9) by using low inhibitor concentrations, however, induced a higher level of active MMP-2 and increased its expression significantly. CONCLUSIONS: The current observations indicate that selective inhibitors of MMPs affect the expression and activity of MMP-2, thus providing clues regarding the differing effects such inhibitors appear to have when applied in vivo. PMID- 12627525 TI - Phase II study of induction chemotherapy with paclitaxel, ifosfamide, and carboplatin (TIC) for patients with locally advanced squamous cell carcinoma of the head and neck. PMID- 12627527 TI - Macroscopic and microscopic analyses of linear enamel hypoplasia in Plio Pleistocene South African hominins with respect to aspects of enamel development and morphology. AB - This study uses macroscopic and microscopic methods to analyze the expression of linear enamel hypoplasia (LEH) in Plio-Pleistocene South African hominins. LEH is a developmental defect of enamel that is used in many anthropological contexts as a physiological stress indicator. Previous research has not settled the question as to whether differences in LEH expression exist between Paranthropus and Australopithecus and if they exist, to what extent these differences might be explained simply by taxonomic differences in enamel development and morphology rather than by differential stress experience. In this study, the analysis of LEH is conducted with respect to differences between Paranthropus and Australopithecus in aspects of enamel development and morphology that are thought to influence LEH expression. Two factors impacting LEH expression are considered: the duration of enamel formation, and the spacing of perikymata. It is predicted that if the first factor strongly influences the expression of LEH, then there should be fewer defects per tooth in Paranthropus because of its abbreviated crown formation spans (and fast extension rates) relative to Australopithecus. It is also predicted that because Australopithecus has more densely packed perikymata in comparable regions of the crown than Paranthropus, this taxon should, on average, have narrower defects than Paranthropus. To address these questions, 200 Australopithecus and 137 Paranthropus teeth were examined for LEH, and the analysis of defect width with respect to perikymata spacing was conducted on tooth impressions examined under a scanning electron microscope using INCA (Oxford Instruments) measurement software. Data support the first prediction: Australopithecus does have significantly more defects per canine tooth than Paranthropus. Data do not support the second prediction in large part because several Australopithecus specimens have wide groove defects in which perikymata are not visible and enamel is irregular. Such wide grooves are not predicted by perikymata spacing such that alternative explanations, including taxonomic differences in ameloblast sensitivity and the duration/severity of disruptions to enamel growth, must be considered. PMID- 12627528 TI - Quantitative analysis of Neanderthal temporal bone morphology using three dimensional geometric morphometrics. AB - The temporal bone is the location of several traits thought to differentiate Neanderthals from modern humans, including some proposed Neanderthal-derived traits. Most of these, however, are difficult to measure and are usually described qualitatively. This study applied the techniques of geometric morphometrics to the complex morphology of the temporal bone, in order to quantify the differences observed between Neanderthal and modern human anatomy. Two hundred and seventy modern human crania were measured, representing 9 populations of 30 individuals each, and spanning the extremes of the modern human geographical range. Twelve Neanderthal specimens, as well as Reilingen, Kabwe, Skhul 5, Qafzeh 9, and 4 Late Paleolithic European specimens, were included in the fossil sample. The data were collected in the form of three-dimensional (3-D) landmark coordinates, and specimen configurations were superimposed using generalized Procrustes analysis. The fitted coordinates were then analyzed by an array of multivariate statistical methods, including principal components analysis, canonical variates analysis, and Mahalanobis D(2). The temporal bone landmark analysis was very successful in separating Neanderthals from modern humans. Neanderthals were separated from modern humans in both the principal components and canonical variates analyses. They were much further in Mahalanobis distances from all modern human populations than any two modern human groups were from each other. Most of the previously described temporal bone traits contributed to this separation. PMID- 12627529 TI - Growth-related shape changes in the fetal craniofacial complex of humans (Homo sapiens) and pigtailed macaques (Macaca nemestrina): a 3D-CT comparative analysis. AB - This study investigates whether macaques and humans possess a common pattern of relative growth during the fetal period. The fetal samples consist of 16 male pigtailed macaques (mean age, 20.5 gestational weeks) and 17 humans (9 males and 8 females; mean age, 29.5 gestational weeks). For each individual, three dimensional coordinates of 18 landmarks on the skull were collected from three dimensional computed tomographic (CT) reconstructed images and two-dimensional CT axial slices. Early and late groups were created from the human (early mean age, 24 weeks, N = 8; late mean age, 34 weeks, N = 9) and macaque samples (early mean age, 17.7 weeks, N = 7; late mean age, 23 weeks, N = 9). Inter- and intraspecific comparisons were made between the early and late groups. To determine if macaques and humans share a common fetal pattern of relative growth, human change in shape estimated from a comparison of early and late groups was compared to the pattern estimated between early and late macaque groups. Euclidean distance matrix analysis was used in all comparisons. Intraspecific comparisons indicate that the growing fetal skull displays the greatest amount of change along mediolateral dimensions. Changes during human growth are primarily localized to the basicranium and palate, while macaques experience localized change in the midface. Interspecific comparisons indicate that the two primate species do not share a common pattern of relative growth, and the macaque pattern is characterized by increased midfacial growth relative to humans. Our results suggest that morphological differences in the craniofacial skeleton of these species are in part established by differences in fetal growth patterns. PMID- 12627530 TI - Differential phenotypic variability among the Apalachee mission populations of La Florida: a diachronic perspective. AB - Phenotypic variability is evaluated in a series of skeletal samples from the Apalachee region of Florida. Based on ethnohistoric evidence, several predictive models for changes in variability are generated. If variability decreases through time, this likely represents the effect of genetic drift in populations experiencing epidemic disease and population loss. If variability increases through time, this suggests that population aggregation or genetic admixture were primary factors shaping the Apalachee population during the mission period. Dental dimensions were collected from a series of precontact (pre-1500), early mission (AD 1633-1650) (San Pedro y San Pablo de Patale), and late mission (post 1657) (San Luis) samples from the Apalachee region and were subjected to univariate and multivariate variability analyses. The results indicate that the late mission San Luis sample was significantly more variable than the Patale or precontact samples; however, the Patale sample exhibited no significant variability change in comparison to the precontact population. This suggests that the missions initially effected limited change in genetic variability in the mission populations. However, San Luis was affected by either admixture or population aggregation to such a degree that the observed variation had increased beyond earlier levels. Given the limited historic evidence for population aggregation at this mission, and the comparatively large resident Spanish population, the increased variability may be indicative of admixture at this mission, and potentially at this mission only. Based on a limited data set, however, it appears that the mission period cannot be typified by a single evolutionary or historic process. PMID- 12627531 TI - Mechanisms of force and power production in unsteady ricochetal brachiation. AB - Brachiators travel by swinging beneath handholds, and it is not obvious how these animals manage to accelerate and decelerate in a horizontal direction, especially when moving rapidly. Most previous analyses focused on brachiation in highly constrained laboratory conditions that induced steady-state locomotion. Emerging understanding of brachiation suggests that much of gibbon locomotory behavior and morphology must be considered within the context of the complexities of the natural environment: the forest canopy is three-dimensional, with high variation in handhold availability and properties. The goal of this paper is to quantify the active mechanisms by which gibbons can dynamically control their velocity. Force production and kinematics were analyzed from a white-handed gibbon Hylabates lar during ricochetal brachiation. Both the mechanisms of force production and power input may be inferred for accelerating and decelerating brachiation by combining force data with kinematics. Examples of steady-state, accelerating, and decelerating ricochetal brachiation are highlighted. Gibbons are able to produce net horizontal impulses by releasing early (resulting in a loss of potential energy, but an accelerating horizontal impulse) or delaying release (associated with an increase in potential energy, and a decelerating horizontal impulse). Torque about the shoulder, leg-lifting (or dropping), and elbow flexing (or straightening) are discussed as potential mechanisms for controlling energy within the brachiating system. Of these possibilities, leg lifting and arm-flexing were observed as mechanisms of adding mechanical energy. Net energy loss, and substantial torques about the shoulder, were not observed. PMID- 12627532 TI - Dynamic plantar pressure distribution during terrestrial locomotion of bonobos (Pan paniscus). AB - We collected high-resolution plantar pressure distributions of seven bonobos during terrestrial bipedal and quadrupedal locomotion (N = 146). Functional foot length, degree of hallux abduction, and total contact time were determined, and plots, showing pressure as a function of time for six different foot regions, were generated. We also studied five adult humans for comparison (N = 13). Both locomotion types of the bonobo show a large variation in plantar pressure distributions, which could be due to the interference of instantaneous behavior with locomotion and differences in walking speed and body dimensions. The heel and the lateral midfoot typically touch down simultaneously at initial ground contact in bipedal and quadrupedal walking of bonobos, in contrast with the typical heel-strike of human bipedalism. The center of pressure follows a curved course during quadrupedalism, as a consequence of the medial weight transfer during mid-stance. Bipedal locomotion of bonobos is characterized by a more plantar positioning of the feet and by a shorter contact time than during quadrupedal walking, according to a smaller stride and step length at a higher frequency. We observed a varus position of the foot with an abducted hallux, which likely possesses an important sustaining and stabilizing function during terrestrial locomotion. PMID- 12627533 TI - Human life histories and the demographic transition: a case study from Finland, 1870-1949. AB - The life histories of humans who were engaged in reproduction during the demographic transition were investigated. It was discovered that these life histories were subject to great changes during the period involving the birth cohorts from the years 1870-1949. Although the number of all and surviving children decreased during this period, the individual fitness values (lambda) of females remained quite even. The lambda values are sensitive not only to reproductive quantity but also to the timing of reproduction. Therefore, the effective change in female fitness during the demographic transition may not be as dramatic as previously thought. When studying the level of selection (or rather the opportunity for selection), it was found that mortality selection steadily decreased to a very low level. However, fertility selection and total selection, which were relatively low for the cohorts 1870-1889, increased before the steep decrease that was detected for the cohorts 1930-1949. The situation reflects the presence of considerable variance in fertility for the cohorts 1890 1929, when the mean fertility was decreasing. A previously found trade-off between female longevity and reproductive success appeared less significant, apparently due to the presence of more plentiful resources and voluntary limitation of reproduction. The deviation from the potential fitness maximization and the presence of subfertility have become prevalent in human populations. PMID- 12627534 TI - Mitochondrial DNA affinities at the Atlantic fringe of Europe. AB - Mitochondrial DNA analysis of Atlantic European samples has detected significant latitudinal clines for several clusters with Paleolithic (H) and Neolithic (J, U4, U5a1, and U5a1a) coalescence ages in Europe. These gradients may be explained as the result of Neolithic influence on a rather homogeneous Paleolithic background. There is also evidence that some Neolithic clusters reached this border by a continental route (J, J1, J1a, U5a1, and U5a1a), whereas others (J2) did so through the Mediterranean coast. An important gene flow from Africa was detected in the Atlantic Iberia. Specific sub-Saharan lineages appeared mainly restricted to southern Portugal, and could be attributed to historic Black slave trade in the area and to a probable Saharan Neolithic influence. In fact, U6 haplotypes of specific North African origin have only been detected in the Iberian peninsula northwards from central Portugal. Based on this peculiar distribution and the high diversity pi value (0.014 +/- 0.001) in this area compared to North Africa (0.006 +/- 0.001), we reject the proposal that only historic events such as the Moslem occupation are the main cause of this gene flow, and instead propose a pre-Neolithic origin for it. PMID- 12627535 TI - Work of nurses caring for HIV-exposed children is largely invisible. PMID- 12627536 TI - Conference speaker will address nursing shortage. PMID- 12627537 TI - ICN and Procter & Gamble will promote clean water. PMID- 12627539 TI - ICN-Merck programme sends key book to African nurses. PMID- 12627538 TI - ICN campaign will help nurses inform patients. PMID- 12627540 TI - [Physician in the fitness studio: possibilities for multiphasic cooperation]. PMID- 12627541 TI - [Nutrition: evaluation of sports beverages and ergogenic substances]. PMID- 12627542 TI - [Shoulder and elbow injuries in ball sports: etiology is often repetitive strain or erroneous load bearing]. PMID- 12627543 TI - [Bicycling: design of the saddle can lead to erection problems]. PMID- 12627544 TI - [Do volleyball players perceive the ball faster than non-athletes?]. PMID- 12627545 TI - [Physical fitness in space: measures to prevent muscular atrophy]. PMID- 12627546 TI - [Medical travel problems with a sports team. What is the role of the physical therapist?]. PMID- 12627547 TI - Does chronic brain edema explain the consequences of cerebral small-vessel disease? PMID- 12627548 TI - Benchmarking the moral decision-making strength of European biotech companies: a European research project. PMID- 12627550 TI - Suspicious minds. PMID- 12627549 TI - Change 'puts patients at risk'. PMID- 12627551 TI - Inquiring mind. Interview by Laura Donnelly. PMID- 12627552 TI - Nutritional genomics. Interview by Deborah Shattuck. PMID- 12627553 TI - Bioethics policy: looking beyond the power of sovereign governments. PMID- 12627554 TI - International bioethics, human genetics, and normativity. PMID- 12627555 TI - The control of genetic research: involving the "groups between". PMID- 12627556 TI - Proposed model ethical protocol for collecting DNA samples. PMID- 12627557 TI - Physician-assisted suicide: should Texas be different? PMID- 12627558 TI - Circumcision and the American Academy of Pediatrics: should scientific misconduct result in trade association liability? PMID- 12627559 TI - Against the golden rule argument against abortion. AB - R.M. Hare and Harry J. Gensler have each argued that abortion can be shown to be immoral by appealing to a version of the golden rule. I argue that both versions of the golden rule argument against abortion should be rejected: each rests on a version of the golden rule which is objectionable on independent grounds, each is unable to support its conclusion when the rule is satisfactorily modified, and each is unable to avoid the implication that contraception is as wrong as abortion and for the same reason. In addition, some further problems particular to each position are identified. PMID- 12627560 TI - Prevention of foodbourne illness and the role of the dietitian. PMID- 12627561 TI - Self-regulation and paragons of virtue: the case of 'fitness to practise'. PMID- 12627562 TI - Skin and bones: post-mortem markets in human tissue. PMID- 12627563 TI - Cloning and the Constitution, cloning and the Constitution, cloning and the Constitution, cloning and .... PMID- 12627564 TI - Endocrine related resources from the National Institutes of Health. PMID- 12627565 TI - Physician assisted death: after the U.S. Supreme Court ruling. PMID- 12627567 TI - Physician assisted suicide--sympathy and skepticism. PMID- 12627568 TI - American Medical Association perspective on physician assisted suicide. PMID- 12627569 TI - Futile care treatment: perspectives from the United States and United Kingdom. PMID- 12627570 TI - What kind of public policy do we need? PMID- 12627571 TI - Engaging patients with psychosis in consultations. To listen or not to listen. PMID- 12627572 TI - Responsible public policy at the end of life. PMID- 12627573 TI - Engaging patients with psychosis in consultations. Cognitive behaviour therapy can help end alienation of psychosis. PMID- 12627574 TI - "She don't want no life support." A summary of Osgood and other developments in Michigan since Martin. PMID- 12627575 TI - Where law and bioethics meet...and where they don't!! PMID- 12627576 TI - Cloning: can it be good for us? An overview of cloning technology and its moral implications. AB - Adequate answers to moral questions about cloning require a working knowledge of the science and technology involved, both present and anticipated. This essay presents an overview of the current state of somatic cell nuclear transfer technology (SCNT), the type of cloning that now permits whole organism reproduction from adult DNA. This essay explains the basic science and technology of SCNT and explores its potential uses. Next, this essay notes remaining scientific obstacles and unanswered moral questions that must be resolved before SCNT can be used for human reproduction. Attention is given to aspects related to cloning for therapeutic and research purposes. PMID- 12627577 TI - Method development and analysis of retail foods and beverages for carotenoid food colouring materials E160a(ii) and E160e. AB - An analytical method using high-performance liquid chromatography with photodiode array detection was developed and applied to the determination of the permitted food colour additives beta-carotene(E160a(ii)) and beta-apo-8'-carotenal (E160e) in foods and beverages. The scope of previously reported methods has been broadened to cover a wide range of retail foods and enzymatic hydrolysis has been used in place of saponification for high-fat samples. Quantitative results (greater then 0.1 mg kg(-1)) are given for the major colour principals trans-beta apo-8'-carotenal and trans-beta-carotene. Semiquantitative results are given for the various cis-isomers of each colorant for which authentic reference standards were not available. The method has been used successfully for the analysis of a wide range of foodstuffs with differing fat content without the need for saponification, except for moderate- to high-fat foodstuffs containing significant levels of emulsifiers, for which it was limited. The results suggest that beta-apo-8'-carotenal (E160e) does not have widespread use in the UK. None of the samples exhibited a total beta-carotene content greater than 20 mg kg(-1) and none of the high-fat samples and only one of the 17 low-fat/beverage samples contained total beta-carotene at levels less than 0.1 mg kg(-1). The total beta carotene contents of the low-fat/beverage samples ranged from 0.4 +/- 0.03 to 8.4 i 0.71 mg kg(-1), and the total beta-carotene contents of the high-fat samples ranged from 0.1 +/- 0.01 (jelly confectionery) to 18.5 +/- 0.98 mg kg(-1) (processed cheese). PMID- 12627578 TI - Much ado about cloning in the public square. AB - The dawnings of the age of human cloning and genetics is shaping lives, ideologies, and social structures. How will we--as individuals and as communities -respond to the possibilities, challenges, and changes of the clone age? This essay invites engagement in communal moral deliberation through broadening conversations about serious matters, including human cloning. A framework that includes important moral markers for significant "kitchen table talk" is offered. Clone age justice is also discussed. Attention is paid to the renewed role of religious voices in the public square. PMID- 12627579 TI - Current medical treatment for tuberculosis. More weight was given to observational studies than randomised controlled trials. PMID- 12627580 TI - Current medical treatment for tuberculosis. Resources used for screening could be spent more usefully elsewhere. PMID- 12627581 TI - Broadening the moral conversation (a response to Margaret McLean). PMID- 12627582 TI - Cloning for therapeutic purposes: ethical and policy considerations. AB - This essay reviews how cloning techniques may be used for therapeutic purposes, analyzes ethical implications, and makes recommendations for public policy discourse. Although cloning may bring many potential benefits, they remain uncertain. Furthermore, human embryo research is morally problematic. Therefore, alternatives to human cloning for therapeutic aims should be sought at present. In addition to central ethical issues, public discourse should maintain an emphasis on the value of the human embryo over scientific expediency, the relativity of health, and the principle of justice. Society should support the laudable mission of medical research, while also attending to the moral concerns often threatened by the promises of scientific progress. PMID- 12627583 TI - Meningoencephalitis due to primary HIV infection. Exceptionalism extends to all "serious infectious diseases". PMID- 12627584 TI - The image of God, the eggs of women, and therapeutic cloning. PMID- 12627585 TI - [Wounds, wound healing, chronic wounds (part 1)]. PMID- 12627586 TI - Human cloning: the role of law. PMID- 12627588 TI - Physical activity: how much is enough? PMID- 12627587 TI - Care of dying patients. Death is not failure but reality. PMID- 12627589 TI - Inversion of emergency pyramid. Inverting pyramids takes time. PMID- 12627590 TI - Inversion of emergency pyramid. "I wanted a thorough checkup". PMID- 12627595 TI - Another putative receptor for hepatitis C virus. PMID- 12627596 TI - Chronic hepatitis B and hepatocarcinogenesis: does prevention of "collateral damage" bring the cure? PMID- 12627597 TI - [Activity of social insurance committee on the standard of surgical facilities]. PMID- 12627598 TI - [Several problems in medical treatment of tumors]. PMID- 12627599 TI - [Applicability of long-term low-dosage macrolide therapy for children with serous otitis media]. PMID- 12627600 TI - [Concerning the etiology of iodine deficiency disorders]. PMID- 12627602 TI - Endocrine-related resources from the National Institutes of Health. PMID- 12627601 TI - [Saliva tests for evaluation of caries risk among school children in Yverdon-les Bains, Vaud]. PMID- 12627603 TI - [Action mechanism of macrolide antibiotics in experimental otitis media]. PMID- 12627605 TI - The role of the generalist obstetrician-gynecologist in the early detection of ovarian cancer. AB - The purpose of this Committee Opinion is to define the role of the generalist obstetrician-gynecologist in the early detection of ovarian cancer. Currently, it appears that the best way to detect early ovarian cancer is for both the patient and her clinician to have a high index of suspicion of the diagnosis in the symptomatic woman. In evaluating symptoms, physicians should perform a physical examination, including a pelvic examination. In pre-menopausal women with symptoms, a CA 125 measurement has not been shown to be useful in most circumstances. In postmenopausal women with a pelvic mass, a CA 125 measurement may be helpful in predicting a higher likelihood of a malignant tumor than a benign tumor, which may be useful in making consultation or referral decisions or both. A woman with a suspicious or persistent complex adnexal mass requires surgical evaluation by a physician trained to appropriately stage and debulk ovarian cancer. Data suggest that currently available screening tests do not appear to be beneficial for screening low-risk, asymptomatic women. An annual gynecologic examination with an annual pelvic examination is recommended for preventive health care. PMID- 12627604 TI - Avoiding inappropriate clinical decisions based on false-positive human chorionic gonadotropin test results. AB - Clinically significant false-positive human chorionic gonadotropin (hCG) test results are rare. However, some individuals have circulating factors in their serum (eg, heterophilic antibodies or nonactive forms of hCG) that interact with the hCG antibody and cause unusual or unexpected test results. False-positive and false-negative test results can occur with any specimen, and caution should be exercised when clinical findings and laboratory results are discordant. Methods to rule out the presence of interfering substances include using a urine test, rerunning the assay with serial dilutions of serum, preabsorbing serum, and using another assay. Physicians must decide whether the risks of waiting for confirmation of results outweigh the risks of failing to take immediate medical action. Patients should be notified if they are at risk for recurrent false positive hCG test results, and this information should be included in the patient's medical record. PMID- 12627606 TI - [Effects of 14-, 15-, and 16-membered ring macrolides on IL-8 gene expression in BET-1A cells]. PMID- 12627607 TI - Chronic fatigue syndrome: symptom subtypes in a community based sample. AB - Most studies of Chronic Fatigue Syndrome (CFS) have been based on patients recruited from primary or tertiary care settings. Patients from such settings might not be typical of patients in the general population. The present investigation involved examining individuals with CFS from a community-based study. A random sample of 18,675 respondents in Chicago were first interviewed by telephone. A group of individuals with chronic fatigue accompanied by at least four Fukuda et al. (1994) symptoms associated with CFS were given medical and psychiatric examinations. From this sample, a physician review group diagnosed individuals with CFS. Those diagnosed with CFS were subclassified based on frequency of symptoms. Important differences emerged on measures of sociodemographics and disability. The implications of these findings and others are discussed. PMID- 12627608 TI - Assessing primary care performance in an obstetrics/gynecology clinic. AB - OBJECTIVES: Young, low-income women often use ob/gyns for routine care. Yet, little is known about the extent to which these settings achieve primary care goals. This study compared primary care performance among young low-income women using ob/gyns versus other settings as their primary care providers (PCP). METHODS: Anonymous interviews with 158 women on Medical Assistance age 18-44 seen at a university ob/gyn clinic. RESULTS: Sixty women (38.0%) used the ob/gyn setting for primary care, 83 (52.5%) use other places and 15 (9.5%) identified no primary care provider. Compared to women using other places for primary care, women who used the ob/gyn setting as their PCP were younger (OR 0.93, p = .04), had less education (OR 3.9, p = .02), and more frequently had small children (OR 2.8, p = .05) and reported illicit drug use (OR 3.1, p = .05). Performance on first contact care, ongoing care, coordination, and family and community issues did not vary by whether the PCP was an ob/gyn versus another provider, though women with an ob/gyn PCP rated their provider significantly higher on cultural competence. Access to advice on general medical prevention topics differed little by type of PCP; however, women who used the ob/gyn clinic as PCP received advice on more reproductive health, behavioral health, and injury prevention topics, with a mean number of preventive topics of 13.0 (SD 3.6) versus 6.3 (SD 4.1) (p < .01). DISCUSSION: Many young women who rely on hospital-based ob/gyn clinics for primary care may benefit from expanded health promotion efforts. The large portion of young, low-income women using ob/gyn settings for primary care may receive primary care of equal or better quality compared to women using other settings. More research is needed to see if these findings are generalizable. PMID- 12627609 TI - A comparison of physical activity in men and women with cardiac disease: do gender roles complicate recovery? AB - Fifty-four male and twenty-six female married cardiac patients were studied for five months post-hospital discharge. Participants completed activity diaries for three days at 5, 10, 15, and 20 weeks. Reported activities were converted to METs. Patients increased activity across the five-month study period. Patients also tended to engage in higher levels of activity in the morning hours. Men and women did not differ in average total METs expended. Comparisons for specific activities indicated that women consistently expended more energy on domestic responsibilities, such as cleaning and laundry. Men engaged more in repairs, yardwork and carrying, but these activities were performed sporadically and involved low METs requirements. The concentrated domestic activity assumed by women early in recovery may pose a risk of complications. PMID- 12627610 TI - Disparities in women's health and health care experiences in the United States and Israel: findings from 1998 National Women's Health Surveys. AB - OBJECTIVE AND METHODS: Using data from bi-national 1998 surveys of adult women in the U.S. and in Israel, this article examines health, access, and care experiences among women in two countries with very different health care systems. We examine how well each country's system serves those vulnerable due to lower socio-economic status. The Israeli health care system-characterized by universal coverage for all its residents-relies on a system of competing health funds that employ many features typical of U.S. managed care plans. The analysis explores the extent to which such a system helps to equalize access experiences with contrasts to the experiences of U.S. women. FINDINGS: We find that U.S. and Israeli women report similar rates of disability and chronic conditions with prevalence of health problems sharply higher for low income and less educated women. We also find disparities in access: women in both countries reported unequal access experiences by education and income. In Israel, these experiences appear to be linked to health plan structural features rather than cost barriers. CONCLUSION: The findings indicate that achieving more equitable access to health care requires attention to non-financial as well as financial barriers to care. Despite the lack of financial barriers to care in Israel, administrative controls typical of managed care organizations appear to make health care systems difficult to navigate for low income and less educated women. The finding that disparities in health persist in a country with universal coverage indicates that improving women's health will require attention to broader social influences on health as well as improving access to health care. PMID- 12627612 TI - Perceptions of power equality among drug-using women. AB - This study examined the associations between perceptions of the power equality in a current sexual relationship and each of several alternative indicators of power in a sample of drug-using women. Using targeted sampling, 262 women were recruited from the streets of Anchorage, Alaska. A battery of face-to-face questionnaires was administered to adult women who gave evidence of recent drug use (track marks or positive urinalysis) and had been recently sexually active. Perceived power equality was associated with physical, verbal, and sexual abuse, self-efficacy, sexual communication, and income for main partners, and the trade of sex and traditionalism of her views on home and family for casual partners. These findings suggest that power is a complex, multifaceted construct, and that type of relationship must be considered when trying to understand the meaning of power within drug-using women's lives. Implications for risk behavior interventions are discussed. PMID- 12627611 TI - The female condom: attitudes and experiences among HIV-positive heterosexual women and men. AB - The female condom is a potentially effective method for the prevention of HIV, other sexually transmitted disease, and pregnancy. This study describes attitudes toward and experiences with the female condom of 89 HIV-positive individuals (n = 56 women; n = 33 men) reporting heterosexual behavior. Qualitative interviews were conducted to inform the design and implementation of a cognitive-behavioral risk-reduction and health-promotion intervention. Most respondents (n = 78) had seen or heard of the female condom. However, relatively few (n = 14 women; n = 5 men) had used it at least once. Reactions from both women and men across user groups, regardless of favorable or unfavorable attitude or experience with the female condom, centered around a similar set of factors: aesthetics, difficulties with the male condom, male partner reaction, beliefs about efficacy, and lack of training. These findings underscore the need for additional research and comprehensive education efforts aimed at both technical use and communication skills-building in order to realize the potential of the female condom as an alternative barrier method. PMID- 12627613 TI - Factors affecting drug and gene delivery: effects of interaction with blood components. AB - Targeted drug delivery systems have been used extensively to improve the pharmacological and therapeutic activities of a wide variety of drugs and genes. In this article, we summarize the factors determining the tissue disposition of delivery systems: the physicochemical and biological characteristics of the delivery system and the anatomic and physiological characteristics of the tissues. There are several modes of drug and gene targeting, ranging from passive to active targeting, and each of these can be achieved by optimizing the design of the delivery system to suit a specific aim. After entering the systemic circulation, either by an intravascular injection or through absorption from an administration site, however, a delivery system encounters a variety of blood components, including blood cells and a range of serum proteins. These components are by no means inert as far as interaction with the delivery system is concerned, and they can sometimes markedly effect its tissue disposition. The interaction with blood components is known to occur with particulate delivery systems, such as liposomes, or with cationic charge-mediated delivery systems for genes. In addition to these rather nonspecific ones, interactions via the targeting ligand of the delivery system can occur. We recently found that mannosylated carriers interact with serum mannan binding protein, greatly altering their tissue disposition in a number of ways that depend on the properties of the carriers involved. PMID- 12627614 TI - Pharmacokinetics of targeting with liposomes. AB - The optimization of drug disposition in the body leads to an increase in its therapeutic effect and to a decrease in adverse effects. Liposomes can serve as a potential drug carrier for achieving this. However, the behavior of a drug carrier system under in vivo conditions is complex. Therefore, a more complete understanding of the pharmacokinetics of liposomes themselves, as well as that of the encapsulated drug, is required. The optimization of the pharmacokinetics of liposomes can be performed by linking a pharmacodynamic model of the free drugs that are encapsulated into liposomes. Sensitivity analysis was applied to optimize the delivery system to maximize the antitumor effect of liposomal doxorubicin (DOX). Advanced technology for ligand-mediated selective targeting and intracellular targeting is also introduced for antitumor agents and for gene delivery systems. PMID- 12627615 TI - US infusion pump market. PMID- 12627617 TI - Surgical treatment of insertional Achilles tendinosis. AB - Most patients with insertional Achilles tendinosis can be managed nonoperatively but those who do not respond may require excision of the diseased tendon. Currently, there are no clinical studies indicating how much of the tendon may be excised without predisposing the patient to Achilles tendon rupture. This chart review reports on 52 heels treated surgically for this condition and followed for a minimum of six months postoperatively. When less than 50% of the tendon was excised (49 heels) patients were immediately mobilized free of a cast. There were two failures using this regimen--one patient with psoriatic arthropathy and another who underwent bilateral simultaneous procedures. We suggest that in selected patients it is safe to proceed with early active mobilization immediately postoperatively when less than 50% of the tendon is resected. PMID- 12627618 TI - Ultrasonography of non-traumatic rupture of the Achilles tendon secondary to levofloxacin. AB - Rupture of Achilles tendon (AT) is an uncommon complication of treatment with fluoroquinolones. We describe a case of bilateral tendinosis and rupture of the right AT in a patient who began levofloxacin treatment for community acquired pneumonia. Sonography showed thickening and hypoecogenicity of both AT and complete rupture and separation of the right Achilles tendon. PMID- 12627619 TI - Outcome of surgical treatment of tarsal tunnel syndrome. AB - One hundred and eight ankles in 72 patients were evaluated from July 1986 to July 1997 with clinical findings and positive electrodiagnostic studies of tarsal tunnel syndrome. Clinical data included physical findings, subjective complaints, duration of symptoms, trauma history, steroid injections, nonsteroidal use and workman's compensation involvement. Associated medical conditions included diabetes, back pain and arthritis. Sixty-two patients underwent tarsal tunnel release, with 13 of them bilateral. There were 44 females and 18 males, 35 right feet and 40 left feet. The average age was 49 years. Preoperative symptom duration was 31 months. Average length of follow-up was 58 months. Average time for return to usual activity was nine months. All patients had at least a 12 month follow-up, and compared with both (Maryland Foot Score) MFS and AOFAS postoperative scores. Preoperative MFS scores obtained prior to 1994, were 61/100 (average), and postoperative MFS scores were 80/100 (average). Postoperative AOFAS scores were 80/100 (average). Patients with symptoms less than one year had postoperative MFS/AOFAS scores significantly higher than those with symptoms greater than one year. The most common surgical findings included arterial vascular leashes indenting the nerve and scarring about the nerve. Varicosities and space occupying lesions were present also. The outcome of surgery was not affected by the presence or absence of trauma. Patients with tarsal tunnel syndrome warrant surgery when significant symptoms do not respond to conservative management. Meticulous surgical technique must be followed. Improvement in foot scores is predictable even when a discrete space-occupying lesion is not present and when symptoms have been present for periods of greater than one year. PMID- 12627620 TI - Tarsal tunnel syndrome associated with an accessory muscle. AB - Between 1986 and 1999, we surgically treated 41 patients (49 feet) with Tarsal Tunnel Syndrome (TTS) in whom seven (eight feet) were associated with an accessory muscle. An accessory flexor digitorum longus muscle was present in six patients, and an accessory soleus muscle was in one patient (both feet). Three of them were males and four females, with the mean age of 33.1 years (12 to 59 years). The mean interval from the onset of symptoms to operation was 7.5 months (range, six to nine months). All patients with an accessory muscle had a history of trauma or strenuous sporting activity. The diagnosis of TTS was made based on physical findings in all the patients (eight feet) and confirmed in five patients (six feet) by electrophysiological examination. Imaging examinations (radiography, ultrasonography, MRI) revealed abnormal bone and soft tissue lesions in and around the tarsal tunnel. Preoperative signs and symptoms disappeared average 4.1 months after decompression of the tibial nerve in addition to excision of the muscle. No functional deficit was observed at final follow-up (24 to 88 months). PMID- 12627621 TI - Stress fractures of the medial great toe sesamoids in athletes. AB - The purpose of this study was to determine whether specific symptoms and findings are present in patients with symptomatic stress fractures of the sesamoids of the great toe and, if so, whether partial sesamoidectomy is sufficient for successful treatment. Five consecutive athletes (five females; mean age 16.8 years [range, 13 to 22 years]) with six feet that were treated for symptomatic stress fractures of the sesamoids of the great toe were included in this study. Four athletes (five feet) performed rhythmic sports gymnastics; the fifth athlete was a long jumper. Some swelling to the forefoot and activity-related pain that increased in forced dorsiflexion, but disappeared at rest was found in all patients. While plain X-rays evidenced fragmentation of the medial sesamoid, MRI (n=2) and frontal plane CT scan (n=3) did not always confirm the diagnosis, but bone scan (n=3) and axial as well as sagittal CT scan were useful to detect the pathology. After failure of conservative treatment measures, surgical excision of the proximal fragment was successful in all patients, and there were no complications. All patients were pain free and regained full sports activity within six months (range, 2.5 to six months). At final follow-up which averaged 50.6 months (range, 20 to 110 months), the overall clinical results were graded as good/excellent in all patients, and there was only one patient with of restriction sports activities. The obtained AOFAS-Hallux-Score was 95.3 (75 to 100) points. Apparently, stress fractures occur more often at the medial sesamoid, and females are mainly involved. When a stress fracture is suspected, bone scan and CT scan are suggested as more reliable in confirming the diagnosis than other imaging methods. When conservative treatment has failed, surgical excision of the proximal fragment is recommended. PMID- 12627622 TI - Suture-endobutton fixation of ankle tibio-fibular diastasis: a cadaver study. AB - Suture-Endobutton fixation is proposed as a minimally invasive, flexible fixation of ankle tibio-fibular diastasis, which would not require routine removal. This study tested the Suture-Endobutton construct in a cadaver syndesmosis injury model and compared this against A.O. syndesmosis screw fixation. Sixteen embalmed cadaver legs were used. Phase one consisted of placing the leg in a jig, generating an external rotation torque and measuring diastasis with increasing intraosseous membrane division. Phase two then compared the Suture-Endobutton construct vs. single four-cortex 4.5 mm A.O. screw fixation. Diastasis increased significantly with increasing intraosseous membrane division (p<0.001). No significant differences were seen in the mean rate of failure between the Suture Endobutton and A.O. screw fixation. However, the Suture-Endobutton did give a significantly more consistent performance; the distribution of standard deviations for A.O. screw fixation was 0.64 mm higher than that for the Endobutton (95% C.I. 0.46 to 0.84). These results show that Suture-Endobutton fixation at least equals the performance of screw fixation and encourages clinical trials in ankle injuries with a syndesmosis diastasis. PMID- 12627623 TI - Operative repair of fourth and fifth toe corns. AB - METHODS: We report a retrospective review of 57 consecutive patients (72 feet) over a period of 20 years who had been treated operatively for either a lateral fifth toe corn or an interdigital corn of the fourth interdigital space more than two years previously. Of these, 51 patients (62 feet) returned for a follow-up evaluation at a minimum of two years (average of over seven years) which included a review of the interval history since the surgery, a physical examination, a radiographic evaluation, and assessment of the patient's satisfaction with the alignment and results of surgery. Treatment of 31 lateral fifth toe corns involved either a lateral condylectomy and flexor tenotomy or a complete condylectomy. Treatment of 31 interdigital corns comprised either a single condylectomy, double condylectomy of adjacent corns, or a complete condylectomy (hammertoe repair) of a symptomatic corn. Treatment in each case was dependent upon the severity of the deformity. RESULTS: There was found to be no significant difference in comparison of the two major groups (interdigital corns and lateral fifth toe corns) with the measurement of the relative length of the fourth and fifth metatarsals, toe malalignment, angulation of the fourth and fifth toes (MTP 4, MTP-5 angles), and the phalangeal-5 angle. Pain was relieved in 58 of 62 feet (93%) and subjective acceptable alignment was achieved in 54 of 62 feet (87%). At final follow-up 53 feet were rated by patients as excellent, seven as good, one as fair, and one as poor. Complications included numbness of the involved digit (six feet). There were two superficial infections. There were two cases of joint instability due to excessive bone resection. Joint stiffness was commonly observed (34/62 feet, 55%), but was not associated with diminished satisfaction at final follow-up. Mild asymptomatic recurrence of a callosity was noted in 10 feet and moderate or severe recurrence was noted in two feet. Dissatisfaction was associated with moderate or severe recurrence. CONCLUSION: In this retrospective study at an average of more than seven years, we achieved a high level of patient satisfaction treating both lateral fifth toe corns and interdigital corns with a partial and/or complete condylectomy, the choice depending upon the magnitude of the deformity and the callus, and the fixed nature of the lesser toe deformity. PMID- 12627624 TI - Injury characteristics and the clinical outcome of subtalar dislocations: a clinical and radiographic analysis of 25 cases. AB - The objective of this study was to determine the mechanisms of injury and pattern of associated foot and ankle injuries and systemic injuries associated with subtalar dislocations, and, correlate these data with the radiographic and clinical/functional outcome of patients after subtalar dislocation. RESULTS: Twenty-five patients with a subtalar dislocation were identified over a seven year period. The mean patient age was 38 years. Males (n=19) comprised 76% of patients, with a mean age of 36 years. High energy mechanisms (motor vehicle accidents, falls) accounted for 68% of subtalar dislocations. Although high energy mechanisms showed a strong trend toward open subtalar dislocations, the association was not statistically significant (p=0.0573, Fisher's exact test). Closed dislocations predominated (75%). Left and right-sided dislocations were nearly equally distributed, even among motor vehicle accidents. Medial dislocations predominated (65%): these were not influenced by mechanism of injury and did not result in statistically lower AOFAS ankle/hindfoot scores. Subtalar dislocation was irreducible (requiring open reduction) in 32%, with higher energy mechanisms of injury being statistically associated with an irreducible subtalar dislocation (p=0.0261, Fisher's exact test). Block to reduction was evenly distributed among soft tissue elements (posterior tibial tendon, flexor hallucis longus tendon, capsule, extensor retinaculum) and osseous elements. Eighty-eight percent of patients incurred concomitant injuries to the foot and ankle (95% of which were closed injuries), namely, the ankle and talus. Systemic injuries occurred in 88% of patients. At a mean follow-up of five years, the mean AOFAS score of the subtalar dislocation side was significantly lower (mean=71 vs. 93, p=0.0007, unpaired Student's t-test). No statistical relation was found between the number of associated extremity injuries and AOFAS score (Spearman correlation coefficient, r=(-)0.236, p=0.331). Radiographic follow-up demonstrated 89% of ankles with radiographic changes (31% symptomatic); however, the majority of these patients (61%) had an associated ankle injury. The subtalar joint demonstrated radiographic changes in 89% of patients, with 63% being symptomatic; 75% of patients with subtalar joint changes incurred a fracture about the subtalar joint at the time of dislocation. Four patients went on to subtalar fusion at an average of 8.8 months post-dislocation. The midfoot showed radiographic changes in 72% of patients, with only 15% of these patients being symptomatic. All patients with midfoot symptoms were well controlled by nonsurgical measures. PMID- 12627625 TI - Transverse plane motion at the ankle joint. AB - The ankle is often considered to have little or no capacity to move in the transverse plane. This is clear in the persistent concept that it is the role of the subtalar joint to accommodate the transverse plane motion of the leg while the foot remains in a fixed transverse plane position on the floor. We present data from noninvasive in vivo study of the ankle subtalar complex during standing internal and external rotation of the leg and study of the ankle subtalar complex during walking. These data reinforce the results of cadaver study and invasive in vivo study of the ankle/subtalar complex. We suggest that the ankle is capable of considerable movement in the transverse plane (generally greater than 15 degrees) and that its role in the mechanism that allows the foot to remain in a fixed transverse plane position on the floor while the leg rotates in the transverse plane, is not simply the transfer of the transverse plane moment to the subtalar joint, but is accommodation of some of the necessary movement. PMID- 12627626 TI - Biomechanical comparison of two fixation methods for first metatarsophalangeal joint arthrodesis. AB - PURPOSE: The aim of this study was to compare the mechanical stability of two methods of fixation for arthrodesis of the hallux metatarsophalangeal joint: 1. a technique using an intramedullary screw and 2. a standard technique using crossed interfragmentary compression screws. METHODOLOGY: The metatarsophalangeal joint was mechanically evaluated in cantilever bending using a servohydraulic testing machine. Differences in stiffness and strength parameters between the two techniques were checked for significance (P<0.05) using a paired t-test. RESULTS: Fixation provided by the intramedullary screw was stiffer and stronger than that from crossed compression screws. CONCLUSIONS: The stronger and stiffer intramedullary screw technique offers mechanical advantages over the crossed interfragmentary screw technique. PMID- 12627627 TI - Avulsion fractures of the medial tubercle of the posterior process of the talus. AB - Avulsion fracture of the medial tubercle of the posterior process of the talus occurs after forceful dorsiflexion-pronation of the ankle. We evaluated five patients who had sustained this fracture while participating in sporting activities. Two patients were correctly diagnosed acutely and treated with immobilization and limited weightbearing. Avulsion fractures in the remaining three patients went undiagnosed acutely. This group was treated with delayed operative excision for persistent posteromedial ankle pain. The patients were evaluated at a mean follow-up of 35 months using the AOFAS Ankle-Hindfoot Scale. The two patients diagnosed and treated acutely achieved excellent results. The three patients with missed fractures did poorly, yet achieved comparable results after late excision. Our results suggest that prompt diagnosis and appropriate management yields reliably good outcomes. Untreated avulsion fractures predictably do poorly. For these patients, late excision can provide significant functional and symptomatic improvement. PMID- 12627628 TI - Endoscopic plantar fascia release: a case series. AB - Plantar fasciitis is a common disabling condition that can be recalcitrant to treatment. Endoscopic Plantar Fascia Release (EPFR) has received greater attention in recent years as a viable, and possibly superior, alternative to established open procedures for the treatment of plantar fasciitis. In a series of 17 patients (17 feet) with follow-up over an average of 16 months, we report a two portal endoscopic technique of partial release of the plantar fascia, as a successful, safe and reliable procedure for the treatment of plantar fasciitis in the patient group selected. The surgical results of 17 EPFR's performed by the same surgeon were reviewed. All patients had preoperative symptoms of subcalcaneal heel pain for greater than 12 months, all having undergone nonsurgical measures for at least 12 months. Of 17 feet, all reported marked improvement of preoperative symptoms within two months. Patients' subjective assessment of the procedure was strongly supportive, 100% totally satisfied or satisfied with only minor restrictions. Postoperatively, 82.4% reported mild or no pain, and 100% had improvement in walking distance. Complications all resolved within two to six months; there were no re-operations and no infections. PMID- 12627629 TI - Review: acute compartment syndrome of the foot. AB - Foot compartment syndrome is a serious potential complication of foot crush injury, fractures, surgery, and vascular injury. The purpose of this article is to summarize and review the existing literature on this entity. Long-term sequelae of foot compartment syndrome (FCS) include contractures, deformity, weakness, paralysis, and sensory neuropathy. These complications are poorly tolerated, and often necessitate multiple procedures for rehabilitation. Therefore, the threshold for considering compartment syndrome and performing fasciotomy must be low to minimize such outcomes. The existence of nine foot compartments and frequent presence of complicating injuries necessitate multi stick needle catheterization for direct measurement of compartment pressures. Fasciotomy is indicated when compartment pressure exceeds 30 mmHg, or if compartment pressure is greater than 10-30 mmHg below diastolic pressure. The approaches for compartment decompression generally include two dorsal incisions for access to forefoot compartments, and one medial incision for decompression of the calcaneal, medial, superficial, and lateral compartments. PMID- 12627631 TI - Multiple computer-automated structure evaluation study of aquatic toxicity. III. Vibrio fischeri. AB - An acute toxicity model was constructed on the basis of 901 chemicals tested for toxicity against the luminescent bacteria Vibrio fischeri (formerly Photobacterium phosphoreum, the Microtox test). The model was created using the Multiple Computer-Automated Structure Evaluation (M-CASE) program. The model can correctly predict acute toxicity for 92% of the compounds with an error averaging 0.55 log units per median effect concentration (EC50). The main toxicophores, corresponding to polar and nonpolar narcosis, and other types of reactive chemicals were identified. PMID- 12627630 TI - Spatial variation of metals and acid volatile sulfide in floodplain lake sediment. AB - In risk assessment of aquatic sediments, much attention is paid to the immobilizing effect of acid volatile sulfide (AVS) on trace metals. The difference of AVS and simultaneously extracted metals (SEM) gives an indication of metal availability. In floodplain sediments, where changing redox conditions occur, AVS may play a major role in determining variation in metal availability. The importance of spatial heterogeneity has been recognized in risk assessment of trace-metal-polluted sediments. However, little is known about spatial variation of available metal fractions. We studied spatial variability of sediment, environmental conditions, total contaminant concentrations, and available metals (as SEM-AVS or SEM-AVS/f(oc)) in a floodplain lake. The top 5 cm of sediment was sampled at 43 locations. Data were analyzed with correlation and principal component analysis as well as with geostatistical methods. Trace metal and SEM concentrations and most sediment characteristics were more or less constant within 10%. In contrast, AVS concentrations were much more variable and showed a strong spatial dependence due to differences in lake depth, total sulfur pools, and redox potential (E(h)), which resulted in crucial differences in trace-metal availability within the lake. The spatial pattern of SEM-AVS deviates from total or normalized trace-metal patterns. This particularly has implications for risk assessment of sediments prone to dynamic hydrological conditions, where AVS concentrations are also variable in time. PMID- 12627632 TI - Studies on bioremediation of polycyclic aromatic hydrocarbon-contaminated sediments: bioavailability, biodegradability, and toxicity issues. AB - The widespread contamination by polycyclic aromatic hydrocarbons (PAHs) has created a need for cost-effective bioremediation processes. This research studied a chronically PAH-contaminated estuarine sediment from the East River (ER; NY, USA) characterized by high concentrations of PAHs (approximately 4-190 ppm), sulfide, and metals and a marine sediment from New York/ New Jersey Harbor (NY/NJH; USA) with only trace quantities of PAHs (0.1-0.6 ppm). The focus was to examine the relationship between bioavailability of PAHs and their biological removal in a slurry system. Freshwater and marine sediment toxicity tests were conducted to measure baseline toxicity of both sediments to amphipods, aquatic worms, fathead and sheepshead minnow larvae, and a vascular plant; to determine the cause of toxicity; and to evaluate the effectiveness of the biotreatment strategies in reducing toxicity. Results showed the ER sediment was acutely toxic to all freshwater and marine organisms tested and that the toxicity was mainly caused by sulfide, PAHs, and metals present in the sediment. In spite of the high toxicity, most of the PAH compounds showed significant degradation in the aerobic sediment/water slurry system if the initial high oxygen demand due to the high sulfide content of the sediment was overcome. The removal of PAHs by biodegradation was closely related to their desorbed amount in 90% isopropanol solution during 24 h of contact, while the desorption of model PAH compounds from freshly spiked NY/NJH sediment did not describe the bioavailability of PAHs in the East River sediment well. The research improves our understanding of bioavailability as a controlling factor in bioremediation of PAHs and the potential of aerobic biodegradation for PAH removal and ecotoxicity reduction. PMID- 12627633 TI - General fugacity-based model to predict the environmental fate of multiple chemical species. AB - A general multimedia environmental fate model is presented that is capable of simulating the fate of up to four interconverting chemical species. It is an extension of the existing equilibrium criterion (EQC) fugacity model, which is limited to single-species assessments. It is suggested that multispecies chemical assessments are warranted when a degradation product of a released chemical is either more toxic or more persistent than the parent chemical or where there is cycling between species, as occurs with association, disassociation, or ionization. The model is illustratively applied to three chemicals, namely chlorpyrifos, pentachlorophenol, and perfluorooctane sulfonate, for which multispecies assessments are advisable. The model results compare favorably with field data for chlorpyrifos and pentachlorophenol, while the perfluorooctane sulfonate simulation is more speculative due to uncertainty in input parameters and the paucity of field data to validate the predictions. The model thus provides a tool for assessing the environmental fate and behavior of a group of chemicals that hitherto have not been addressed by evaluative models such as EQC. PMID- 12627634 TI - Predicted and measured levels of azinphosmethyl in the Lourens River, South Africa: comparison of runoff and spray drift. AB - Runoff and spray drift are important sources of nonpoint pesticide pollution in surface waters, but few studies have directly compared these routes of input in an exposure assessment scenario. To this end, a runoff formula suggested by the Organization for Economic and Cooperative Development (Paris, France) and basic drift values (95th percentiles) were integrated into a geographical information system (GIS) to predict runoff and spray drift-related loading of azinphosmethyl (AZP) in the Lourens River (LR), South Africa. The GIS-integrated calculations were first validated in the tributaries of the river, where measured loads were well predicted for both runoff (r(20 = 0.95; p < 0.0001; n = 9) and spray drift (r(2) = 0.96; p = 0.0006; n = 8). Through extrapolation to the catchment scale containing 400 ha of orchards, the GIS-integrated calculations predicted similar loads of AZP as measured in the Lourens River mainstream for six runoff (between a factor of 1.03 and 1.86 lower) and six spray drift (between a factor of 1.1 and 2.4 higher) events. Mean measured loads per event were significantly (p = 0.004) higher for runoff (27.8 +/- 19.1 g) than for spray drift (0.69 +/- 0.32 g). Based on long-term meteorological data and average application regimes, runoff leads to a higher annual load (47.6 g) than spray drift (5.5 g) in the Lourens River. Runoff is clearly a more important source of nonpoint pollution in the studied catchment, and mitigation strategies should focus first on addressing this aspect on a catchment scale and second on addressing problem areas on a subcatchment scale. PMID- 12627635 TI - The use of carbon thirteen nuclear magnetic resonance spectra to predict dioxin and furan binding affinities to the aryl hydrocarbon receptor. AB - Four spectroscopic data-activity relationship (SDAR) models for polychlorinated dibenzofurans (PCDFs) and dibenzodioxins (PCDDs) binding to the aryl hydrocarbon receptor (AhR) have been developed based on simulated 13C nuclear magnetic resonance (NMR) data. Models were developed using discriminant function analysis of the compounds' spectral data. An SDAR model with two classifications for 26 PCDF compounds had a leave-one-out (LOO) cross-validation accuracy of 89%. A two classification SDAR model for 14 PCDD compounds had LOO cross-validation accuracy of 95%. A two-classification SDAR model combining 14 PCDD and 26 PCDF compounds had LOO cross-validation accuracy of 88%, while a four-classification SDAR model based on the same 14 PCDD and 26 PCDF compounds had LOO cross-validation accuracy of 92%. We used each appropriate SDAR model to classify 41 PCDD and/or 121 PCDF compounds with unknown binding affinities to the AhR. The SDAR models provide a rapid, simple, and valid way to model the PCDF and PCDD binding activity in relation to the AhR. PMID- 12627636 TI - Biodegradation during contaminant transport in porous media: 6. Impact of sorption on coupled degradation-transport behavior. AB - Bioavailability is one of the critical factors influencing the biodegradation and bioremediation of organic compounds. The bioavailability of many organic contaminants is controlled in part by the nature, magnitude, and rate of sorption/desorption processes. This study investigates the impact of sorption and associated retardation on the bioavailability and biodegradation of aromatic hydrocarbons during transport in porous media. Miscible-displacement experiments were conducted using naphthalene and 2-naphthol as the model sorbing compounds and salicylate, a degradation product of naphthalene, as a nonsorbing reference compound. Two porous media were used, one (Eustis soil, FL, USA) with moderate sorption capacity and one (quartz sand) with no measurable sorption of the compounds. The porous media were sterilized and inoculated with Pseudomonas putida RB1353, an organism that degrades naphthalene and its derivatives. The biodegradation and transport of all three substrates in quartz sand were significantly influenced by microbial lag, the effects of which were observed within two to three pore volumes (3-4.5 h). This was also true for salicylate transport in the Eustis soil system. Conversely, biodegradation lag effects were not observed for naphthalene or 2-naphthol in the Eustis soil system. In addition, the masses of naphthalene and 2-naphthol degraded were significantly greater for the Eustis soil system compared to the quartz sand system. As noted previously, naphthalene and 2-naphthol were sorbed by the Eustis soil but not the quartz sand, while salicylate was not sorbed by either media. These results indicate that the increased residence time associated with sorption of naphthalene and 2-naphthol by Eustis soil enhanced overall biodegradation and obviated the impact of lag on observed transport behavior. PMID- 12627637 TI - Measuring the bioavailability of two hydrophobic organic compounds in the presence of dissolved organic matter. AB - Bioavailability of benzo[a]pyrene (BaP) and 3,3',4,4'-tetrachlorobiphenyl (TCB) was studied in natural lake water containing dissolved organic matter (DOM). Lake water was diluted to give a dissolved organic carbon (DOC) range of 1 to 20 mg/L. Partition coefficients for the model compounds were assessed at different DOM concentrations and over time with three different methods, namely equilibrium dialysis and reverse-phase and liquid-liquid extraction. In addition, biological partition coefficients were estimated from the difference in the bioconcentration of the model compounds in Daphnia magna in the presence and absence of DOM. Results showed that bioavailability of the model compounds was reduced by the presence of DOM. The equilibrium dialysis method gave the best estimates for bioavailability of the model compounds when compared with biologically determined values. Both the reverse-phase and the liquid-liquid extraction overestimated the bioavailable fraction. The more pronounced overestimation of bioavailable fraction of TCB suggested that the sorption of TCB was not only lower but the interaction was also weaker than that of BaP. Increasing DOM concentration produced lower partition coefficients and the effect seemed to be more pronounced when measured by the reverse-phase and the extraction methods. PMID- 12627639 TI - Identification of phototransformation products of prednisone by sunlight: toxicity of the drug and its derivatives on aquatic organisms. AB - Solar simulator irradiation of an aqueous suspension of prednisone, a widely prescribed drug, produces seven photochemical derivatives. The compounds have been identified on the basis of their physical features. All the chemicals have been tested to evaluate their toxic effects on freshwater organisms from different trophic levels. The rotifer Brachionus calyciflorus and two crustaceans, the cladoceran Daphnia magna and the anostracan Thamnocephalus platyurus, were used to perform acute toxicity tests. Chronic toxicity tests have been performed on the alga Pseudokirchneriella subcapitata (formerly known as Selenastrum capricornutum) and the crustacean Ceriodaphnia dubia. The results showed low acute and chronic toxicity of prednisone. Some of the photoproducts had high toxic effects on C. dubia. PMID- 12627638 TI - Role of metalloporphyrin core metals in the mediated reductive dechlorination of tetrachloroethylene. AB - A promising approach to abiotically dechlorinate a variety of chlorinated organic contaminants under reducing conditions is to utilize porphyrins or other tetrapyrrole macrocycles as electron transfer mediators/shuttles for catalyzing their reduction. In this study, various experimental approaches were used to elucidate the role of porphyrin core metals in the reductive dechlorination of tetrachloroethylene (PCE). The importance of specific core metals for the reactivity of a porphyrin and its mediated reaction mechanisms was demonstrated by inserting different metals into metallo tetrakis (N-methyl-4-4 pyridiniumyl) porphyrin (TMPyP). No PCE dechlorination was observed when the free-base (i.e., no core metal) and iron core metal forms of TMPyP were utilized. When using nickel or cobalt TMPyP, reductive dechlorination of PCE occurred but appeared to follow different pathways for the two metals based on product analyses. Physical (e.g., steric) considerations suggest that direct contact between a porphyrin core metal and PCE may be limited and therefore that the entire metalloporphyrin molecule should be viewed as a functional system in which the organic macrocycle has an active part in reductive dechlorination reactions. This view is supported by the fact that slight changes in the functional groups on a porphyrin macrocycle, particularly those far removed from the core metal itself, greatly affected the reactivity and mechanism of the porphyrin. Solution conditions also had a major effect on porphyrin reactivities, to the extent that a nonreactive metalloporphyrin could be activated merely by adjusting the pH of the solution or by adding a small amount of cosolvent. The collective results of this study suggest that fine tuning of naturally occurring metalloporphyrin complexes and/or their environments can enhance the catalyzed detoxification of chlorinated contaminants in many natural and engineered environmental systems. PMID- 12627640 TI - Biotransformation of 3,5-dibromo-4-hydroxybenzonitrile under denitrifying, Fe(III)-reducing, sulfidogenic, and methanogenic conditions. AB - Bromoxynil (3,5-dibromo-4-hydroxybenzonitrile) is a halogenated aromatic nitrile herbicide used on a variety of crops for the postemergence control of annual broad-leaved weeds. The anaerobic biodegradability of bromoxynil and its aerobic transformation product, 3,5-dibromo-4-hydroxybenzoate, were examined in enrichment cultures established with anaerobic sediment under denitrifying, Fe(III)-reducing, sulfidogenic, and methanogenic conditions. Bromoxynil (100 microM) was depleted in 20 to 30 d in the methanogenic, sulfidogenic, and Fe(IIi) reducing enrichments but was stable under denitrifying conditions. The 3,5 dibromo-4-hydroxybenzoate (100 microM) was depleted within 20 to 35 d under all four anaerobic conditions. Both compounds were stable in sterile controls. Bromoxynil and 3,5-dibromo-4-hydroxybenzoate were readily utilized upon respiking of the cultures. During utilization of bromoxynil, stoichiometric release of bromide was observed with transient accumulation of metabolites identified as bromocyanophenol, cyanophenol, and phenol. Bromoxynil heptanoate and octanoate were rapidly hydrolyzed to bromoxynil, which was further degraded. These results indicate that bromoxynil and 3,5-dibromo-4-hydroxybenzoate are degraded under different anaerobic conditions. Anaerobic degradation of bromoxynil proceeds via reductive debromination to 4-cyanophenol, which is further transformed to phenol and can ultimately be degraded to carbon dioxide. PMID- 12627641 TI - Contamination of fish in streams of the Mid-Atlantic Region: an approach to regional indicator selection and wildlife assessment. AB - The extent of contamination of fish in the Mid-Atlantic Region was evaluated as part of the U.S. Environmental Protection Agency's Monitoring and Assessment Program's regional assessment in 1993 through 1994. Fish assemblages from wadeable streams were dominated by small, short-lived fishes (e.g., minnows, darters, and sculpins) that were more widely distributed and abundant than large fishes typically chosen for tissue contaminant studies (e.g., trout, black bass, sunfish, common carp). Chemical concentrations in whole-fish homogenates exceeded detection limits for mercury, DDT, and polychlorinated biphenyls (PCBs) in 75 to 100% of the stream length assessed using small fishes and 84 to 100% of the stream length assessed using large fishes. Wildlife values (WVs) representing a threshold for toxic effect were developed to allow examination of the spatial extent of potential risk to piscivorous wildlife. For mercury, DDT, dieldrin, and chlordane, estimates of the regional extent of streams where fish contaminant concentrations exceeded the WVs were greater when based on small fishes than on large fishes. However, within the distribution of stream lengths assessed using small and large fishes, the percentage of stream kilometers exceeding the WVs were quite similar. Our data demonstrate that the greater abundance and distribution of small, short-lived fishes provide greater estimates of regional extent of contamination for first- through third-order streams and can be used for regional assessments of potential exposure and effects in wildlife. PMID- 12627642 TI - Developmental toxicity of in ovo exposure to polychlorinated biphenyls: I. Immediate and subsequent effects on first-generation nestling American kestrels (Falco sparverius). AB - We determined that in ovo exposure to polychlorinated biphenyls (PCBs) alters growth of first-generation nestlings during and one year after parental exposure. Captive American kestrels (Falco sparverius) laid eggs with environmentally relevant total PCB levels (34.1 microg/g whole-egg wet wt) when fed PCB-spiked (Aroclor 1248, 1254, and 1260) food (7 mg/kg body wt/ d) for 100 d in 1998. In 1999, the same adults laid eggs with estimated total PCBs of 29.0 microg/g. Nonsurviving PCB-exposed chicks were small (mass, bones) in 1998. Survivors showed a strong sex-specific growth response (mass, bones) compared to respective sex controls: Only female hatchlings were larger, and only male nestlings had longer feathers (1998); maximal growth and bone growth rates also differed (males were advanced, faster; females delayed, slower) (1999); and male nestlings fledged earlier and were smaller, while females were larger (1998, 1999). However, regardless of sex, PCB-exposed nestlings generally grew at faster rates in both years. In 1998, greater contaminant burdens and toxic equivalent concentrations in sibling eggs were associated with nestlings being lighter, having longer bones and feathers, and growing at faster rates (mass, bone) for females but slower rates (mass) for males. Both physiological-biochemical and behavioral changes are likely mechanisms. This study supports and expands on the Great Lakes embryo mortality, edema, and deformities syndrome: While PCB exposure alters nestling size, maximal growth and growth rates also change immediately, are sustained, and are sex specific. PMID- 12627643 TI - Copper bioaccumulation by the freshwater snail Lymnaea peregra: a toxicological marker of environmental and human health? AB - Snails (Lymnaea peregra) were exposed to both low and high concentrations of copper(as copper nitrate) undercontrolled conditions and then were sacrificed and dissected; various tissues/organs were removed and subsequently analyzed by atomic absorption spectrophotometry to determine both the copper concentration and the nature of localization. At low concentrations, bioaccumulation was evident in various tissues/organs; a dose response occurred in tissues derived from both the head and foot. At high concentrations, the copper concentrations of the liver-like body and kidney became massively enhanced. There was some variability in the ranking of tissues/organs with regard to copper bioaccumulation--this was predominantly linked to the copper concentration to which the snails were exposed. Mechanisms of copper bioaccumulation are reviewed and comparisons with other organisms made. PMID- 12627644 TI - Predicting sediment toxicity using logistic regression: a concentration-addition approach. AB - The question posed in this article is how useful the chemical concentration measurements for predicting the outcome of sediment toxicity tests are. Using matched data on sediment toxicity and sediment chemical concentrations from a number of studies, we investigated several approaches for predicting toxicity based on multiple logistic regression with concentration-addition models. Three models were found to meet criteria for acceptability. The first model uses individual chemicals selected using stepwise selection. The second uses derived variables to reflect combined metal contamination, polycyclic aromatic hydrocarbon (PAH) contamination, and the interaction between metals and PAHs. The third and final model is a separate species model with derived variables. Overall, these models suggest that toxicity may be correctly predicted approximately 77% of the time, although prediction is better for samples identified as nontoxic than for those known to be toxic. PMID- 12627645 TI - Comparison of the response of three microalgae species exposed to elutriates of estuarine sediments based on growth and chemical speciation. AB - The elutriate sediment toxicity test (ESTT) provides a measure of the amount of a substance that is exchanged between the sediment and the aqueous phase during resuspension processes such as floods or dredging operations. This study used ESTT with two complementary aims: a comparison of the elutriates of two estuarine sediments (anaerobic muddy [A] and aerobic sandy [B]) in terms of toxicity and a comparison of the response of three different microalgae (Emiliania huxleyi (coccolithophore), Dunaliella minuta (green alga), and Phaeodactylum tricornutum (diatom)) to each elutriate in terms of growth, heavy metals uptake, and organic ligands release or uptake in order to find eventual differences of sensitivity. The interpretation of the results was based on chemical speciation in the culture media. Both elutriates, particularly A, were much richer than seawater (control medium) in some heavy metals and organic ligands able to bind strongly heavy metals. Elutriate A slightly inhibited P. tricornutum growth but stimulated growth of E. huxleyi and D. minuta. Elutriate B stimulated the growth of the three algal species. Therefore, the diatom behaved differently from both the coccolithophore and the green alga. Strong complexation of trace metals by organic ligands could be the cause of absence of the metallic toxicity of the elutriates. Growth inhibition of P. tricornutum in elutriate A could be caused by ammonia-N and/or organic compounds. The concentration of the organic ligands decreased markedly (about 75%) in both elutriates after 10 d of incubation in contrast to the control culture, where their concentration increased about 50% because of exudation. This phenomenon was interpreted to result from ligand uptake by the algae, free or as metal complexes. This work demonstrated that beside the evaluation of toxicity of free heavy metals to alga species, the organic ligands must not be ignored. Depending on the amount of ligand present, the toxicity can be reduced (sequestration) or enhanced (better availability through uptake of metal-ligand complexes). Since the applied ESTT is a standard procedure (U.S. Environmental Protection Agency) for the evaluation of dredged material proposed for ocean disposal, it is necessary to discuss results obtained during toxicity tests with such elutriates in detail. PMID- 12627646 TI - Relationships between mercury body concentrations, standard metabolic rate, and body mass in eastern mosquitofish (Gambusia holbrooki) from three experimental populations. AB - Eastern mosquitofish (Gambusia holbrooki) were sampled from three experimental populations (two Hg-exposed populations and one reference population) to determine whether transgenerational exposure (lifelong exposure of multiple generations) to Hg adversely affects standard metabolic rate (SMR). Mosquitofish subjected to lifelong Hg exposure accumulated significant concentrations of Hg in their tissues compared to fish from the reference population (mean: 3.89-4.13 vs 0.08 microHg/g wet mass, respectively). Less than 10% of the variability in Hg tissue concentrations could be explained by fish body mass, likely because of the short life span and/or dietary habits of this species. Despite the high body burdens of Hg in exposed fish, we found no significant difference in SMR among individuals from Hg-exposed or reference populations. Our findings contrast recent laboratory work describing elevated SMR in mosquitofish exposed to 100 microg/L dissolved inorganic Hg for 48 h. To account for contrasting results between studies, we hypothesize that acute exposure to dissolved inorganic Hg damages gill epithelium, resulting in increased metabolic rate, but that lifelong Hg exposure via trophic uptake of methyl mercury does not affect fish respiratory structures. Alternative hypotheses include the possibility that G. holbrooki is a species that can tolerate high body burdens of Hg or that more than four years of genetic isolation during Hg exposure (8-12 generations) resulted in selection for Hg-tolerant or -resistant individuals. PMID- 12627647 TI - The role of ultraviolet-adaptation of a marine diatom in photoenhanced toxicity of acridine. AB - Cultures of the marine diatom Phaeodactylum tricornutum were grown under laboratory light with a different fraction of ultraviolet radiation (UV) to study the potential role of photoadaptation in determining the sensitivity to photoenhanced toxicity of acridine. In short-term experiments, a higher acridine concentration was needed to inhibit the photosynthetic electron flux, monitored with chlorophyll a fluorescence, in algae exposed to fluorescent light (low UV) than to mercury light (high UV), consistent with the expected role of UV. The two types of light in long-term exposures led to changes in the pigment composition and photosystem I (PS I) to photosystem II (PS II) stoichiometry to optimize the utilization of fluorescent and mercury light. Despite the adaptation of the photosynthetic apparatus to a small fraction of UV, long-term exposure to mercury light did show a constant sensitivity of the photosynthetic efficiency of P. tricornutum to the phototoxic acridine. It is concluded that the prime receptor of photoenhanced toxicity may be unrelated to the photosynthetic machinery. PMID- 12627648 TI - Essential and desirable characteristics of ecotoxicity quantitative structure activity relationships. AB - Quantitative structure-activity relationships (QSAR) developed and applied in the prediction of ecotoxic potencies far out number those in other areas, such as health effects. There are yet to be any formal guidelines for the development of ecotoxicological QSARs. Despite this, the depth and breadth of our knowledge of QSARs as they apply to ecotoxicology, especially short-term aquatic toxicity, allow for the formulation of characteristics that appear to be essential and/or desirable for high-quality QSARs. The three components of a QSAR are the biological activity, the property/structural descriptors, and the statistical methodology. Problems may arise from all three components and may be compounded by interactions between them. In an effort to minimize any tribulations associated with development and application of ecotoxic QSARs, a number of essential or desirable characteristics have been identified. Ecotoxicological data used in formulating the QSAR must be reliable, of high quality, and reflect a well-defined and continuous endpoint; this dataset should be diverse both in terms of potency and chemical structure (i.e., property). Descriptors used in formulating the QSAR should be of high quality, reproducible, of a number and type consistent with the endpoint being modeled, and when possible allow for a mechanistic interpretation of the QSAR. The statistical process used in formulating a QSAR should be as rigorous as possible, appropriate for the endpoint being modeled, and allow for the development of as easily interpretable (i.e., transparent) QSARs as possible. The resultant QSAR should be validated, only used within the descriptor space and chemical domain of the model, and relied on in relation to the total weight of evidence; precision of the QSAR and expectations from its application need to be related to the error in the original ecotoxicological and descriptor measurements. Finally, development of QSARs should be through the interaction of a group of multidisciplinary experts. PMID- 12627650 TI - Toxicity of 19 adjuvants to juvenile Lepomis macrochirus (bluegill sunfish). AB - Nineteen adjuvants, many used as surfactants for aquatic herbicide applications, were applied in static bioassay to bluegill sunfish (Lepomis macrochirus) for 96 h to determine median lethal concentrations (LC50). Surfactants are added to the tank mix as a percentage (v/v) of the total volume, in contrast to herbicide application rates, which are usually expressed in kilograms per hectare. Two ethoxylated tallow amine products were the most toxic, having LC50 values of 1.6 and 2.9 ppm (all values v/v). Seven alcohol/glycol-based surfactants had 96-h LC50 values of 4.0 to 11.6 ppm (mean = 7.9 ppm). The polysiloxane- or silicone based surfactants had toxicities of 18.1 to 29.7 ppm (mean = 24.7). Two limonene based products had LC50 values of 10.2 and 30.2 ppm. A methylated seed oil with emulsifier had a LC50 of 53.1 ppm. Two acid/buffer utility adjuvants had LC50 values of 60.8 and 221 ppm. To compare the relative safety of the tested surfactants, we assumed maximum label rate applications to 1 m deep water with uniform mixing. This comparison of relative safety is based on mortality to 50% of the test organisms and does not imply application rates that would not result in any mortality. The two ethoxylated tallow amines, neither used or recommended for aquatic applications, had a relative safety factor of 12.6 or less. Relative safety factor varied from 6.2 to 20.4 for the seven alcohol/glycol surfactants, 38.4 to 63.2 for silicone-based products, 5.5 to 16.1 for limonene products, 113 for methylated seed oil, and 132.2 to 315.7 for acid/buffer utility adjuvants. When used according to label recommendations under normal use conditions, these adjuvants should not be present in acutely toxic concentrations; however, the most toxic adjuvants in very shallow water (< 10 cm) would be toxic to bluegill sunfish that did not move to deeper water to avoid lethal concentrations. PMID- 12627649 TI - Perfluorooctane sulfonic acid in bib (Trisopterus luscus) and plaice (Pleuronectes platessa) from the Western Scheldt and the Belgian North Sea: distribution and biochemical effects. AB - A biomonitoring campaign was conducted in the Belgian North Sea and in the Western Scheldt (The Netherlands) with the primary goal to assess perfluorooctane sulfonic acid (PFOS) contamination and distribution in different biota. This study covers the results obtained for bib (Trisopterus luscus) and plaice (Pleuronectes platessa) and includes the assessment of some stress-related biochemical endpoints. Analysis of liver and muscle PFOS concentrations of both species provided evidence for the existence of a PFOS pollution gradient along the Western Scheldt with higher levels at the upstream locations and a lower degree of PFOS pollution at the marine locations. Cellular necrosis was studied by measuring aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels in the serum. Serum ALT but not serum AST was shown to correlate positively with the PFOS liver concentration in bib (r = 0.44, p < 0.05), indicating that PFOS might contribute to the induction of hepatic damage in bib in the area of study. Analysis of total carbohydrate, lipid, and protein content of bib liver tissue revealed a positive correlation between the protein content and the PFOS liver concentration (r = 0.55, p < 0.01). Whether this is due to induction of compensatory mechanisms, detoxification, or repair processes remains unclear. PMID- 12627651 TI - Failure of the biotic ligand and free-ion activity models to explain zinc bioaccumulation by Chlorella kesslerii. AB - Zinc accumulation by Chlorella kesslerii (Chlorophycee) was studied for [Zn2+] ranging from 4 pM to 1 mM. A first-order uptake flux as predicted by the free ion activity model (FIAM) and the biotic ligand model (BLM) was not observed. Furthermore, when algae were preconditioned in slightly limiting (10 pM) versus optimal (1 nM) free zinc concentrations, the internalization flux increased and was nearly constant over the range of [Zn2+] examined. This observation was attributed to the synthesis of membrane-bound zinc transporters. Neither solution Zn chemistry nor surface-bound Zn was a good predictor of Zn uptake fluxes. Several hypotheses were examined to explain the failure of the steady-state uptake models. Although zinc excretion and a Zn diffusion limitation were observed, neither result could explain the majority of observed deviation from the models. Bioaccumulation experiments performed with inhibitors (vanadate and carbonyl cyanide m-chlorophenylhydrazone [CCCP]) demonstrated clearly that zinc transport is an energy-dependent process in Chlorella kesslerii. The presence of an active transport signifies that Zn uptake may function independently of the electrochemical Zn gradient and that, in some cases, both uptake fluxes and receptor-bound Zn may be independent of solution chemistry. PMID- 12627652 TI - Effects of sublethal concentrations of atrazine and nitrate on metamorphosis of the African clawed frog. AB - Tadpoles of the African clawed frog (Xenopus laevis) were exposed to sublethal concentrations of atrazine (0, 40, and 320 microg/L) and nitrate (0, 37, and 292 mg/L) from feeding stage to metamorphosis. A 3 x 3 factorial design was used to identify both single and interactive effects. At metamorphosis, tadpole weight, snout-vent length (SVL), and hematocrit were determined. Mean mortality was greater in tanks receiving 320 microg/L atrazine; nitrate had no effect on mortality. Significant differences for all mean traits at metamorphosis occurred among atrazine treatments; higher atrazine exposure increased time to metamorphosis and decreased weight, SVL, and hematocrit. Nitrate treatments were not significantly different. Significant interaction tests between atrazine and nitrate occurred for weight and SVL at metamorphosis; the specific type of interaction varied among treatments. Assuming an additive mixture model, at low atrazine (40 microg/L), the addition of 37 mg/L nitrate produced SVL values less than expected (a synergistic effect) while the addition of 292 mg/L nitrate yielded SVL values greater than expected (an antagonistic effect). A similar response was noted for tadpoles in the 320-microg/L atrazine treatments. These results indicate that environmentally realistic concentrations of atrazine exert a negative impact on amphibian metamorphosis. Also, this study suggests that mixtures of agricultural chemicals, even if sublethal, may exert negative and not necessarily consistent mixture effects. PMID- 12627653 TI - Hepatic microsomal cytochrome P450 enzyme activity in relation to in vitro metabolism/inhibition of polychlorinated biphenyls and testosterone in Baltic grey seal (Halichoerus grypus). AB - Among other factors, cytochrome P450 (CYP) enzyme activity determines polychlorinated biphenyl (PCB) bioaccumulation, biotransformation, and toxicity in exposed species. We measured the oxidative metabolism in vitro of 12 PCB congeners, representing structural groups based on the number and position of the chlorine atoms, by the hepatic microsomes of one Baltic grey seal (Halichoerus grypus). Microsomal metabolism was observed for several PCBs with vicinal H atoms exclusively in the ortho and meta positions and without any ortho-Cl substituents (CB-15 [4,4'-Cl2] and CB-77 [3,3',4,4'-Cl4]), vicinal meta and para-H atoms (CB 52 [2,2',5,5'-Cl4], and -101 [2,2',4,5,5'-Cl5]) or with both characteristics in combination with either only one ortho-Cl (CB-26 [2,3',5-Cl3], CB-31 [2,4',5 Cl3]) or two ortho-Cl substituents (CB-44 [2,2',3,5'-Cl4]). To allocate PCB biotransformation to specific CYPs, the inhibitive effect of compounds with known CYP-specific inhibition properties was assessed on in vitro PCB metabolism and on regio- and stereospecific testosterone hydroxylase activities. Metabolic inhibition was considered relevant at concentrations < or = 1.0 microM because these inhibitors became decreasingly selective at higher concentrations. At < 1.0 microM, ellipticine (CYPIAI/2 inhibitor) selectively inhibited CB-15, -26, -31, and -77 metabolism, with no significant inhibition of CB-44, -52, and -101 metabolism. Inhibition of CB-52 and -101 metabolism by chloramphenicol (CYP2B inhibitor) started at 1.0 microM and maximized at about 100% at 10 microM. Ketoconazole (CYP3A inhibitor) appeared to selectively inhibit CB-26, -31, and 44 metabolism relative to CB-15, -77, and -52 at concentrations < or = 1.0 microM. Major testosterone metabolites formed in vitro were 2beta-(CYP3A), 6beta- (CYP3A, CYPIA), and 16beta- (CYP2B) hydroxytestosterone and androstenedione (CYP2B, CYP2C11). The CYP forms indicated are associated with the specific metabolism of testosterone in laboratory animals. Inhibition of 2beta- and 6beta hydroxytestosterone formation at ellipticine and ketoconazole concentrations < or = 1.0 microM suggested that both inhibitors were good substrates of CYP3A-like enzymes in grey seal. Chloramphenicol (model for CYP2B) is apparently not a good inhibitor of CYPI A and CYP3A activities in grey seal because the chemical did not inhibit any metabolic route of testosterone at concentrations from 0.1 to 10 microM. Our findings demonstrated that at least CYP1A- and CYP3A-like enzymes in the liver of grey seals are capable of metabolizing PCBs with ortho-meta and/or meta-para vicinal hydrogens. A CYP2B form might also be involved, but this could not be proven by the results of our experiments. Defining the profiles of CYP enzymes that are responsible for PCB biotransformation is necessary to fully understand the bioaccumulation, toxicokinetics, and risk of PCB exposure in seals and other free-ranging marine mammals. PMID- 12627654 TI - Biomass growth, micronucleus induction, and antioxidant stress enzyme responses in Vicia faba exposed to cadmium in solution. AB - Biomass growth, micronucleus induction, and antioxidative stress enzymes (superoxide dismutase, peroxidase, glutathione reductase, and catalase) were investigated simultaneously in the Vicia faba plant exposed to cadmium in solution. The biomass lowest-observed-effect concentration (LOEC) value was 2,000 microM Cd2+. In the shoots, enzymic activities increased without concentration response relationships. In the roots, after an initial increase, activities of all enzymes showed negative concentration-response relationships. A significant increase in micronucleus induction was observed at 20 microM Cd2+. Regarding sensitivity, our results showed that biomass endpoint was less sensitive than micronucleus induction, which was less sensitive than antioxidative enzyme activities. The increase of antioxidant stress enzyme activities in response to cadmium exposure may be taken as evidence for an enhanced detoxification capacity of V. faba plants toward reactive oxygen species (and derivatives) that might be generated in the stressed plants. Concomitant micronucleus induction may be also interpreted as a consequence of oxidative stress, upholding the view that cadmium induced DNA damage is, to some extent, via generation of reactive (intermediate) oxygen species. PMID- 12627655 TI - Photoenhanced toxicity of aqueous phase and chemically dispersed weathered Alaska North Slope crude oil to Pacific herring eggs and larvae. AB - The photoenhanced toxicity of weathered Alaska North Slope crude oil (ANS) was investigated in the eggs and larvae of Pacific herring (Clupea pallasi) with and without the chemical dispersant Corexit 9527. Oil alone was acutely toxic to larvae at aqueous concentrations below 50 microg/L total polycyclic aromatic hydrocarbons (tPAH), and median lethal (LC50s) and effective concentrations (EC50s) decreased with time after initial oil exposure. Brief exposure to sunlight (approximately 2.5 h/d for 2 d) significantly increased toxicity 1.5- to 48-fold over control lighting. Photoenhanced toxicity only occurred when oil was present in larval tissue and increased with increasing tPAH concentration in tissue. Ultraviolet radiation A (UVA) treatments were less potent than natural sunlight, and UVA + sunlight caused greater toxicity than sunlight alone. The toxicity of chemically dispersed oil was similar to oil alone in control and UVA treatments, but oil + dispersant was significantly more toxic in the sunlight treatments. The chemical dispersant appeared to accelerate PAH dissolution into the aqueous phase, resulting in more rapid toxicity. In oil + dispersant exposures, the 96-h no-observed-effect concentrations in the UVA + sunlight treatment were 0.2 microg/L tPAH and 0.01 microg/g tPAH. Exposure of herring eggs to oil caused yolk sac edema, but eggs were not exposed to sun and UVA treatment did not cause phototoxicity. These results are consistent with the hypothesis that weathered ANS is phototoxic and that UV can be a significant and causative factor in the mortality of early life stages of herring exposed to oil and chemically dispersed oil. PMID- 12627656 TI - Analysis of monitoring data of pesticide residues in surface waters using partial order ranking theory. AB - In this investigation, a new and simple way to analyze, interpret, and generalize monitoring data of occurrence of pesticide active ingredients in surface waters was developed. The occurrence is quantified using the variables frequency of detection and the concentration level. These two parameters are associated with basically different ecotoxicological effects; for example, a high frequency of detection may be related to bioaccumulation problems, while the level of concentration also controls the acute toxicological effects. The active ingredients were ranked on the basis of the monitoring data in relation to both the frequency of finding and concentration level using the concept of partial ordered sets. The resulting rankings was correlated with other rankings based on descriptors such as sprayed area, applied dose, adsorption to soil organic carbon, vapor pressure, and soil dissipation half-life. A similarity index was applied in order to compare the ranking of the monitoring data with the ranking of the descriptors. It is shown how partial order theory can be used to evaluate the relevance of every single descriptor. The dosage is found to be the most important descriptor, followed by the sprayed area and the adsorption to organic carbon ending up a very close similarity between, respectively, the rankings using monitoring data and rankings using these three descriptors. PMID- 12627657 TI - Genetic structure and mtDNA diversity of Fundulus heteroclitus populations from polycyclic aromatic hydrocarbon-contaminated sites. AB - Genetic structure and diversity of mummichog (Fundulus heteroclitus) populations were investigated using mitochondrial DNA (mtDNA) sequences. Forty-six haplotypes were identified among 208 mummichog from the Elizabeth and York Rivers in Virginia, USA. No evidence of decreased gene or nucleotide diversity for mummichog from polycyclic aromatic hydrocarbon (PAH)-contaminated sites was observed. However, based on mtDNA data from 17 sites, a significant correlation (Mantel analysis, p = 0.035) was noted between genetic distance (F(ST)) and PAH concentration but not between genetic distance and geographic distance. Mummichog from the most heavily PAH-contaminated site, Atlantic Wood (AW), were genetically distinct from those of other Elizabeth River sites. At AW, high frequencies of several divergent haplotypes were observed that were more closely allied to the northern mummichog than to the more abundant southern form in the Chesapeake Bay. These data suggested that a locally stable population existed at the AW site. This conclusion is consistent with the observation that mummichog from the AW site display enhanced tolerance to PAH contamination relative to mummichog from noncontaminated sites. Conclusions about gene diversity and the correlation between genetic distance with site differences in PAH concentrations were also consistent with those from tandem genetic analyses based on allozymes. PMID- 12627658 TI - Population-level responses to long-term cadmium exposure in two strains of the freshwater gastropod Biomphalaria glabrata: results from a life-table response experiment. AB - A life-table response experiment was conducted to ascertain the demographic effects of low-level cadmium exposure on two strains, BS90 and NMRI, of the freshwater gastropod Biomphalaria glabrata. Snails were exposed to cadmium continuously from the embryonic stage through adulthood. Results indicated that cadmium significantly affected a number of individual-based parameters, including %hatch, juvenile survival, and adult survival in both strains. Also, fecundity and time to maturity were significantly affected in the NMRI strain. A stage based, deterministic, population model indicated that population growth rate (lambda) was significantly affected by cadmium. Elasticity analysis indicated that juvenile survival, in general, had the greatest contribution to lambda. Decomposition analysis indicated that the effects of cadmium on the juvenile stage in BS90 and the embryonic stage in NMRI contributed most to cadmium-induced changes in lambda. The BS90 strain was more sensitive to cadmium toxicity than NMRI. Moreover, the two strains differed in pattern of response with different aspects of their respective life histories contributing to cadmium-induced changes in lambda. Comparisons were also made between the main model, based on a Z transformation of the life-cycle graph, and more commonly used matrix models. PMID- 12627659 TI - Thrombosis and thrombophilia: diagnosis and management. PMID- 12627660 TI - Introduction to thrombosis proficient and cost-effective approaches to thrombosis. AB - Thrombosis is the most common single cause of death in the United States. More than 2 million people die each year of arterial or venous thrombosis or its consequences, and a similar number experience nonfatal thrombosis-deep vein thrombosis, nonfatal pulmonary embolus, nonfatal cerebrovascular thrombosis, transient cerebral ischemic attacks (40% of patients will have a fatal or nonfatal cerebrovascular thrombosis within 1 year), nonfatal coronary artery thrombosis, retinal vascular thrombosis, and other nonfatal thrombotic episodes. Yet many, if not most, episodes of thrombosis can be prevented by appropriate primary antithrombotic therapy, and most recurrences can be prevented by the appropriate choice of secondary therapy. PMID- 12627661 TI - Prothrombin G20210A mutation, antithrombin, heparin cofactor II, protein C, and protein S defects. AB - These defects are not as common as factor V Leiden, but they are more common than many other hereditary procoagulant defects. The incidence of the prothrombin gene (G20210A) mutation is not yet known with certainty, but it may approach or even exceed that of factor V Leiden. These defects also seem less common than hereditary sticky platelet syndrome; however, they are all common enough that they always should be considered in any individual with unexplained thrombosis and should be part of the work-up for patients with thrombotic disorders. Of the defects discussed herein, prothrombin G20210A mutation seems, thus far, to be more common than AT, protein C, protein S, or HC-II defects. Assessment of prothrombin gene mutation should be part of the primary evaluation of patients with unexplained thrombosis. PMID- 12627663 TI - Sticky platelet syndrome and thrombocythemia. AB - Platelets are intimately involved in the pathogenesis of thromboembolic disorders, especially arterial forms of thrombosis. Although most arterial thromboses develop on the basis of endothelial injuries, some do not. In these instances "hyperactive" platelets could be the cause. Hyperaggregable platelets have been described in association with a number of acquired disease entities whereby the cause-and-effect relationship is unclear. In contrast, the sticky platelet syndrome is a congenital, autosomal dominant disorder, characterized by hyperaggregable platelets in response to ADP, epinephrine, or both. Patients usually seek treatment for transient or permanent arterial vascular occlusions. These are often precipitated by stressful events. Treatment with low-dose aspirin (81 mg/day) reverses clinical symptoms and hyperaggregability in the laboratory. PMID- 12627662 TI - Activated protein C resistance (FV(Leiden)) and thrombosis: factor V mutations causing hypercoagulable states. AB - The integrity of the vascular system is of prime importance for survival. Therefore, several emergency and repair systems safeguard the circulatory system. Multiple processes jointly limit vascular damage and blood loss. In this article, the authors focus on the protein C anticoagulant pathway and the role of activated protein C resistance in thrombotic disease, and they discuss the involvement in thrombosis of mutations other than the Arg506Gln mutation in the gene encoding for factor V. PMID- 12627664 TI - Hyperhomocysteinemia and thrombosis. AB - Homocysteine remains an enigmatic marker for vascular disease. Studies have shown hyperhomocysteinemia is a risk factor for VTE, cerebrovascular disease, and coronary artery disease. This relationship, however, has not been consistently corroborated by studies of patients with genetic polymorphisms that alter homocysteine metabolism. Studies at the molecular level reveal interactions between homocysteine, the endothelium, and the clotting system. Further investigation at the basic science level is needed to determine whether homocysteine is a marker of vascular injury and thrombotic potential or whether it plays a pathogenic role. Preliminary trials with vitamins clearly show that safe, inexpensive treatment can lower homocysteine levels. The clinical impact on decreasing vascular disease, however, has yet to be shown. Until there is evidence that treatment improves outcomes, testing for homocysteine and treating hyperhomocysteinemia will be a debatable issue. A series of vitamin intervention trials begun in 1997 will enroll tens of thousands of patients (Table 1) and will, it is hoped, provide the necessary information for developing evidence based guidelines. PMID- 12627665 TI - Hereditary and acquired defects in the fibrinolytic system associated with thrombosis. AB - The fibrinolytic system plays a pivotal role in the regulation of hemostasis and the prevention of thrombosis. There are no drugs that will increase the plasma fibrinolytic activity for a lasting duration to prevent thrombotic events effectively. Despite the ability of vasoactive agents such as nicotinic acid and metformin to release PA from the vessel wall, this therapeutic effect has not been evaluated adequately. The PAs are short-acting and indicated only for thrombolysis and not for prophylaxis. Future directions are directed at finding, agents that can enhance plasminogen activator release or inhibit PAW-1 activity. As there are multiple factors involved in the pathogenesis of thrombosis, there are a number of conditions in which abnormal fibrinolysis is only a contributory factor. Examples are seen in pregnancy, especially during puerperium, when the thromboembolic risk is at its highest. The levels of inhibitors of fibrinolysis. both PAI-1 and PAI-2, are also at their highest. Another example was seen recently in the antiphospholipid syndrome, where antibodies against Annexin II, a receptor for tPA, were found to be higher than in healthy controls. Thus, a thorough investigation into other hereditary and acquired risk factors for thrombosis is recommended. PMID- 12627666 TI - Antiphospholipid thrombosis syndromes. AB - Antiphospholipid antibodies are associated strongly with thrombosis and are the most common of the acquired blood protein defects causing thrombosis. Although the precise mechanisms whereby antiphospholipid antibodies alter hemostasis to induce a hypercoagulable state remain unclear, numerous theories, as previously discussed, have been advanced. The most common thrombotic events associated with ACLAs are deep vein thrombosis and pulmonary embolus (type I syndrome), coronary or peripheral artery thrombosis (type II syndrome), or cerebrovascular/retinal vessel thrombosis (type III syndrome), and occasionally patients present with mixtures (type IV syndrome). Patients with type V disease are those with antiphospholipid antibodies and RMS. It is as yet unclear how many seemingly normal individuals who may never develop manifestations of antiphospholipid syndrome (type VI) harbor asymptomatic antiphospholipid antibodies. The relative frequency of ACLAs in association with arterial and venous thrombosis strongly suggests that they should be looked for in any individual with unexplained thrombosis; all three idiotypes (IgG, IgA, and IgM) should be assessed. Also, the type of syndrome (I-VI) should be defined, if possible, because this identification may dictate both the type and the duration of immediate and long term anticoagulant therapy. Unlike those patients with ACLAs, patients with primary LA-thrombosis syndrome usually have venous thrombosis. Because the aPTT is unreliable in patients with LA (prolonged in only approximately 40%-50% of patients) and usually is not prolonged in patients with ACLAs, definitive tests, including ELISA for ACLA, the dilute Russell's viper venom time for LA, hexagonal phospholipid-neutralization procedure, and B-2-GP-I (IgG, IgA, and IgM) should be ordered immediately when suspecting antiphospholipid syndrome or in individuals with otherwise unexplained thrombotic or thromboembolic events. If these test results are negative, subgroups also should be assessedin the appropriate clinical setting. Most patients with antiphospholipid thrombosis syndrome will fail to respond to warfarin therapy, and except for retinal vascular thrombosis, may fail some types of antiplatelet therapy, so it is of major importance to make this diagnosis so patients can be treated with the most effective therapy for secondary prevention-LMWH or unfractionated heparin in most instances and clopidogrel in some instances. PMID- 12627667 TI - Disseminated intravascular coagulation current concepts of etiology, pathophysiology, diagnosis, and treatment. AB - The pathophysiologic mechanisms and clinical and laboratory manifestations of DIC are complex, partly because of inter-relationships within the hemostasis system. Only by clearly understanding these extraordinarily complex pathophysiologic inter-relationships can the clinician and laboratory scientist appreciate the divergent and wide spectrum of often-confusing clinical and laboratory findings in patients with DIC. Many therapeutic decisions to be made are controversial and lack validation. Nevertheless, newer antithrombotic agents and agents that can block, blunt, or modify cytokine activity and the activity of vasoactive substances seem to be of value. The complexity and variable degree of clinical expression suggest that therapy should be individualized depending on the nature of DIC, the patient's age, etiology of DIC, site and severity of hemorrhage or thrombosis, and hemodynamics and other appropriate clinical parameters. PMID- 12627668 TI - Current concepts in the diagnosis and management of thrombotic thrombocytopenic purpura. AB - Thrombotic thrombocytopenic purpura is a multisystem disease characterized by thrombocytopenia, hemolytic anemia, renal failure, fever, and neurologic abnormalities. Plasma exchange has revolutionized the outcome of this entity from a once fatal disease to a disease that potentially is cured or has prolonged remission. The understanding of the pathophysiology of TTP continues to evolve. Recently, investigators showed that a deficiency in a specific plasma protease responsible for cleaving vWf plays a crucial role in the familial form of TTP. This explains in part why patients usually respond to plasma exchange therapy. The identification of a mutation in a specific gene that belongs to the metalloproteinase family located at chromosome 9q34 could have important therapeutic implications. TTP can be induced by certain drugs, especially immunosuppressants, in the setting of bone marrow and solid organ transplantation. This disease also has been described in association with HIV, pregnancy, cancer, and chemotherapy. TTP remains an ideal example of how knowledge about the etiology of a disease can improve therapeutic interventions. PMID- 12627669 TI - Coumarin therapy in thrombosis. AB - Oral anticoagulants are the most commonly used agents in the long-term prophylaxis and treatment of arterial and venous thrombotic disorders. As new and expanded indications for their use, such as the prevention of recurrent myocardial infarction or the treatment of systemic embolism in atrial fibrillation, are developed, the use of oral anticoagulants is rising. Also, in North America, oral anticoagulants are used commonly for preventing venous thromboembolism following orthopedic surgery. This article reviews the pharmacology of warfarin sodium, the most commonly used oral anticoagulant in North America, and discusses practical aspects of the use of this agent in thrombotic disorder management. PMID- 12627670 TI - Thromboprophylaxis and thrombosis in medical, surgical, trauma, and obstetric/gynecologic patients. AB - The International Consensus and the ACCP Sixth Consensus had a great impact on the clinical acceptance of LMWHs. These recommendations have been instrumental in initiating further clinical trial to answer key questions regarding thromboprophylaxis and in setting a new standard for patient care. Also, the key to cost containment in management of DVT/PE is to (1) define the etiology (blood coagulation protein or platelet defect), institute appropriate long-term therapy as indicated, and assess appropriate family members as indicated if a hereditary defect is found and (2) use LMWH as inpatient management. saving a minimum of 210,000.00 dollars per 1000 patients simply from cost savings of recurrence, saving 17 lives per 1000 patients, and saving exorbitant costs of care for patients with recurrence and development of chronic venous insufficiency. The use of outpatient LMWH will save 4,900,000.00 dollars per 1000 patients if applied to the 70% of patients with DVT who fit the criteria of no comorbid condition requiring hospitalization and who arrive early enough to allow a diagnosis to be sent home or hospitalized for 24 hours or less. The simple defining of defects leading to unexplained thrombosis will add another 3,000,000.00 dollars in savings per 1000 patients with DVT and approximately 350,000.00 dollars per 100 patients with thrombotic stroke. In those with transient ischemic attacks, defining the defect and instituting appropriate antithrombotic therapy, thereby potentially saving approximately 30% from developing a thrombotic stroke, amounts to approximately 350,500.00 dollars (= 30% of 1,168,500.00 dollars) in savings per 100 patients. PMID- 12627672 TI - Thrombolytic therapy: clinical applications. AB - The therapeutic use of thrombolytic agents is the result of the increasing understanding of the pathophysiologic mechanisms underlying normal and deranged thrombosis and fibrinolysis. Plasminogen activators capable of increasing the production of plasmin exhibit considerable efficacy in the treatment of a variety of arterial and venous thrombotic disorders. The ideal thrombolytic agent has not been developed, but the desired clinical result of rapid opening of the thrombosed vessel without reocclusion, without activation of systemic fibrinogenolysis, and without a risk of hemorrhage are defined. Clinical studies clearly demonstrate that the addition of a variety of adjunctive agents to available thrombolytics enhances benefit without inordinate risk. The addition of intravascular angioplasty and stenting to thrombolysis increases the potential long-term benefit. Newer thrombolytic agents and new protocols for the use of existing therapies offer the promise of saving many who would otherwise succumb to coronary or cerebral arterial thrombosis or to venous thromboembolism. PMID- 12627671 TI - Heparin-induced thrombocytopenia, paradoxical thromboembolism, and other adverse effects of heparin-type therapy. AB - This article discusses the common and uncommon side effects of heparin and heparin-like therapy. Most attention is devoted to heparin-induced thrombocytopenia, as it is the most clinically aggressive adverse reaction. Additionally, less common and several newly reported side effects are discussed. PMID- 12627673 TI - Heparin, low-molecular-weight heparins, and heparin pentasaccharide: basic and clinical differentiation. AB - As a result of advanced technology, dramatic developments in the area of new anticoagulant and antithrombotic drugs appear to have made a profound impact on the use of LMWHs. Furthermore, because porcine mucosal heparin is used for the preparation of these agents, it is likely that alternative drugs with comparable pharmacologic and clinical efficacy are sought. Antithrombin drugs such as argatroban and hirudin are already approved for alternative management of heparin compromised patients. Their efficacy in other indications is less superior. The development of specific anti-Xa drugs is slow. Although these agents may inhibit factor Xa and thrombin generation, none of them are capable of mimicking the polytherapeutic effects of LMWHs and thus can only be given in drug combinations. Synthetic and recombinant protein-derived anti-tissue factor agents have also been developed. These drugs only inhibit the tissue factor-mediated process and are limited in their therapeutic spectrum. Plasma-derived and recombinant serine protease inhibitors (serpins) are also available for the management of thrombotic and inflammatory disorders, but these agents cannot be given subcutaneously. Furthermore, because they are proteins, antibodies to these agents are generated. Nucleic acid derivatives (natural and synthetic aptomers) are developed for intravenous administration, but they are relatively weak antithrombotic agents. Dermatans, heparans, and chondroitin sulfates represent nonheparin GAGs, and, in mono-compositional and polycompositional form, these drugs are mainly used for the intravenous management of DVT prophylaxis. They can be given to patients who are heparin compromised. Synthetic heparinomimetics include heparin consensus binding oligosaccharides and synthetic oligosaccharides with non-serpin affinity. In addition, binding oligosaccharides are conjugated with antithrombin agents to mimic the anti-Xa/anti-IIa activities of heparin. Biotechnology using bacterial and yeast cultures, aqua cultures for marine products, and plant carbohydrates have been the focus of developing heparin analogues. Development of these agents is in the early phase; however, it is likely that this approach may provide a reasonable alternative to LMWHs. Despite these developments, it is unlikely that any of these drugs will have a profound impact on the use of LMWHs in the near future. Unfractionated heparin and LMWHs collectively represent an important group of polypharmacologic drugs without which the management of thrombosis and vascular disorders would not be possible. The continual development of LMWHs in expanded indications did not comprise the use of unfractionated heparin in surgical and interventional cardiovascular indications. Ever since their introduction in the 1980s, the use of LMWHs has continually increased. This is primarily because of expanded indications and growing awareness among the clinicians. It is likely that once an antidote is developed and additional information is available on the mechanism of action of LMWHs, these drugs may gradualty be used for surgery patients. Despite these developments, it is likely that unfractionated heparin will continue to be used for specific indications. Drug combinations with heparins may necessitate dose adjustments, but it is unclear whether unilateral reduction of heparins will be optimal. The coming years will provide useful clinical and applied data on the improved use of unfractionated heparin. LMWHs, and pentasaccharide in the management of thrombotic and cardiovascular disorders. In addition, use of these drugs will be extended to many conditions, including cancer, inflammation, sepsis, and autoimmune diseases. Polytherapeutic approaches emphasizing LMWHs as primary and secondary drugs will also have an impact on the management of thrombotic and nonthrombotic disorders. Ultra-LMWHs and synthetic heparinomimetics, such as fondaparinux, that exhibit a narrow pharmacologic spectrum will only be useful in specific indications and in combination with other drugs. PMID- 12627674 TI - The Lund lecture--The future of the criminal justice system. PMID- 12627675 TI - The single joint expert. AB - The new concept in civil litigation is that of the single joint expert (SJE). Is it a good concept or a bad concept? What are the merits and demerits? What changes in practice are emerging? PMID- 12627676 TI - Tightropes and tragedies: 25 years of Tarasoff. AB - In 1976 the California Supreme Court decided the case of Thrasoff v Regents of the University of California. That case has had very significant effects on psychiatric practice on both sides of the Atlantic, and its very name is instantly recognizable to most psychiatrists. Yet the details of the case are often misunderstood or misquoted; as a legal precedent it was rapidly superceded, and is now largely redundant. Twenty-five years on it seems appropriate to revisit the case itself, and its impact on contemporary psychiatric practice. English law has yet to impose a similar duty, but two leading cases raising similar issues are also described. PMID- 12627677 TI - A novel PCR method for identifying plankton in cases of death by drowning. AB - We present a new PCR method for identifying plankton in cases of death by drowning. We designed four primer pairs for chlorophyll-related genes of Euglena gracilis (EG) and Skeletonema costatum (SK), which are commonly distributed in water. The primers were selected from sequences coding chloroplast/chlorophyll apoprotein of EG (EG1 and EG2) and fucoxanthin-chlorophyll a/c harvesting protein of SK (SK1 and SK2). With EG1 or EG2, up to 2 fg of EG-DNA was identified, and 0.2 pg of SK-DNA was detectable with SK1 or SK2. No PCR products were amplified from green vegetables (komatsuna, spinach, parsley) or human DNA with the four primer pairs. Regardless of the origin, seawater or fresh water, most diatoms were detectable with primer pairs of EG1 and EG2. With SK1, only Centrales diatoms were identified, and five diatom strains originating from seawater were detectable with SK2. EG1 and EG2 gave rise to PCR products from most water samples. By using Percoll, plankton was easily isolated from human tissue or blood samples and good results of PCR analysis were obtained in cases of death by drowning. PMID- 12627678 TI - Control and restraint techniques in the management of violence in inpatient psychiatry: a critical review. AB - Violence has long been a matter of concern in inpatient psychiatry. While research suggests that training in physical restraint techniques can reduce the number and severity of violent incidents and assault-related injuries, the recent Cochrane Review is critical of the methodological inadequacies which characterise these studies. This paper considers issues pertinent to understanding research in this controversial area, critically evaluates research on the effectiveness of the predominant approach to training in physical restraint in the UK, and examines some of the methodological problems inherent in this research. PMID- 12627679 TI - Evaluating alleged drinking after driving--the hip-flask defence. Part 2. Congener analysis. AB - The second part of this review describes the principles and practice of forensic congener analysis as an alternative way to evaluate claims of drinking alcohol after driving. Congener analysis was developed, perfected and practised in Germany as a way to evaluate hip-flask defences. This kind of defence challenge arises frequently when the drunk driving suspect is not apprehended at the wheel and especially after hit-and-run incidents. Besides ethanol and water, alcoholic beverages contain trace amounts of many other low-molecular substances, known collectively as the congeners, which impart the characteristic smell and taste to the drink. Importantly, the congener profile can be used to identify a particular kind of alcoholic beverage. Forensic congener analysis entails making a qualitative and quantitative analysis of ethanol, methanol, n-propanol and the isomers of butanol in blood and urine from the apprehended driver and comparing the results with the known congener profile of the alcoholic beverage allegedly consumed after driving. Interpreting the results of congener analysis requires knowledge about the absorption, distribution and elimination pattern of the congener alcohols, including their oxidation and conjugation reactions, and any metabolic interactions with ethanol. Complications arise if drinks with widely different congener profiles are consumed or if the same beverage was ingested both before and after driving. Despite these limitations, congener analysis can furnish compelling evidence to challenge or support claims of drinking alcohol after driving. PMID- 12627680 TI - The nature and management of aggressive incidents in a medium secure unit. AB - The study describes and analyses incident reports over a three-year period in a medium secure unit. A low threshold of reporting was encouraged and substantial numbers of 'minor' incidents were reported. The frequency of incidents peaked in the evenings. The use of seclusion and manual restraint is reported and these were used more frequently where staff were the target of aggression. PMID- 12627681 TI - The use of Section 136 of the Mental Health Act 1983 in a family of rural English police forces. AB - Section 136 of the Mental Health Act 1983 is used by police officers to detain persons who they feel might be suffering with mental disorder until a formal Mental Health Act assessment can be undertaken. Previous studies have shown that the outcomes of these assessments result in remarkably different rates of subsequent hospital admissions. Within a rural setting it has also been shown that the rate of use of Section 136 varies considerably. This study examines the use of Section 136 within a family of eight police forces that have been matched to ensure that they cover similar populations. The results show that there are considerable variations in the use of Section 136, with the Devon and Cornwall region using the section over two and a half times the mean for the group. Possible reasons for this discrepancy are discussed. PMID- 12627682 TI - Probation orders with conditions of psychiatric treatment: a descriptive study. AB - Although probation orders with conditions for psychiatric treatment (POPTs) have been in existence for many years there is very little literature relating to their use and almost none from a psychiatric perspective. All patients who were subject to, or had recently completed, a POPT in Leicestershire were identified and information on them collected from psychiatric and probation records. Of 33 POPTs identified, psychiatric information was found for 31 and probation for 23. The most striking feature of the sample was its great heterogeneity in terms of psychiatric diagnoses and index offences. Worryingly, a number of serious offences did not lead to charges or convictions and there was evidence of poor communication between and within agencies, particularly psychiatric services. Our recommendations for improvement mirror those of other studies in the area, namely, better training of the staff from the multiple agencies involved, improved communication, better documentation and ongoing audit to prevent the situation continuing. PMID- 12627683 TI - 1. The myth of child rape as a cure for HIV/AIDS in Transkei: a case report. AB - South Africa has one of the highest cases of HIV/AIDS infection in Africa, and Transkei, a former black homeland, now a part of the Eastern Cape Province, is one locality with a large number of HIV/AIDS sufferers. The unemployment level is very high and crime, including child rape, is very common. This report presents the case of a victim of rape, a nine-year old female child who was brought to the Umtata General Hospital, a victim of the mistaken belief that sex with a virgin will cure an HIV-infected person or AIDS sufferer of his illness. The alleged rapist was an HIV-positive uncle of the child. The myth of the 'HIV/AIDS virgin cure' is prevalent in the community. The history, physical examination and laboratory investigations of this case are given. A conclusion is drawn and preventive methods are suggested. PMID- 12627684 TI - Sudden death and heart weight. PMID- 12627686 TI - Extending the hand of knowledge: promoting mobile technologies. AB - The widespread use of handheld computers and other mobile devices in the healthcare environment and their potential for providing access to information has prompted health sciences librarians everywhere to learn more about this technology. Early in 2001, the Health Sciences Library (HSL) at the University of North Carolina at Chapel Hill began exploring ways to support mobile computing. This paper describes the four basic approaches taken by the librarians that helped establish the HSL as a leader in the area of mobile technologies. PMID- 12627688 TI - The benefits of library liaison programs for small libraries: an overview. AB - Library liaison programs are commonly used and provide a successful framework for communication in academic libraries. Liaison programs, whereby librarians are formally designated as the primary contact between the library and one or more departmental or administrative units, are proven to improve the transfer of information between the library and users, to improve the quality of collections and services, and to enhance the library's image. Previously published literature on liaison programs is primarily devoted to large-scale liaison programs in academic settings, the market where this model is commonly employed. Small hospital and other smaller libraries are nearly absent in the literature, reflecting the low level of liaison use in the smaller library setting. This article invites hospital and other smaller libraries to explore the liaison model by presenting common liaison goals and activities that are not only pertinent to, but also scalable, adaptable, and adoptable by smaller and larger libraries alike. PMID- 12627689 TI - ACTIS and ATIS: new resources for AIDS information, part two. AB - The HIV/AIDS Treatment Information Service (ATIS) is a Web-based resource for federally approved treatment guidelines for HIV and AIDS. ATIS and ACTIS (AIDS Clinical Trials Information Service) are managed by the National Library of Medicine. Background information, search hints, and a content description of ATIS are provided. PMID- 12627687 TI - Implementing PDA technology in a medical library: experiences in a hospital library and an academic medical center library. AB - Personal digital assistants (PDAs) have grown from being a novelty in the late 1990s to an essential tool for healthcare professionals in the 2000s. This paper describes the experiences of a librarian who implemented PDA technology first in a hospital library, and then at an academic medical center library. It focuses on the role of the library in supporting PDA technology and resources. Included are programmatic issues such as training for library staff and clinicians, and technical issues such as Palm and Windows operating systems. This model could be used in either a hospital or academic health sciences library. PMID- 12627690 TI - U.S. Government statistics on the World Wide Web. AB - The U.S. Government produces and provides massive amounts of statistics. Government initiatives have made much of the statistical information emanating from its various federal agencies, departments, and offices digitally accessible on the Internet. Assisting users in locating specific statistics can be one of the most challenging duties librarians have. This article provides a guide to government sources that will help librarians locate health statistics for researchers, health professionals, health consumers, and others. Some selected sites may be an endpoint of the search, and others may offer a solid beginning to a complex answer. Links are also provided to gateway Web sites, where a search strategy utilizing some of the tips discovered while compiling this list can be used to help promote successful outcomes. As always, it is recommended to use evaluative criteria to assess information quality as well as integrity of the data. PMID- 12627691 TI - Health smart cards: merging technology and medical information. AB - Smart cards are credit card-sized plastic cards, with an embedded dime-sized Integrated Circuit microprocessor chip. Smart cards can be used for keyless entry, electronic medical records, etc. Health smart cards have been in limited use since 1982 in Europe and the United States, and several barriers including lack of infrastructure, low consumer confidence, competing standards, and cost continue to be addressed. PMID- 12627692 TI - Beyond the literature: bioinformatics training for medical librarians. AB - As genetics and molecular biology progressively impact on society and the practice of medicine, medical librarians must prepare themselves to deal with this new arena of information. The Eskind Biomedical Library at Vanderbilt University Medical Center has developed a bioinformatics training program for its librarians involving subject knowledge, literature evaluation, and database searching techniques. This program can serve as a model for other libraries. PMID- 12627693 TI - Our multicultural society: implications for pediatric dental practice. PMID- 12627694 TI - Our multicultural society: implications for pediatric dental practice. Keynote speaker, 17th annual symposium, Denver, Colorado, Saturday, May 25, 2002. PMID- 12627695 TI - Dental disease prevalence, preventon, and health promotion: the implications on pediatric oral health of a more diverse population. PMID- 12627696 TI - Multicultural influences on child-rearing practices: implications for today's pediatric dentist. PMID- 12627697 TI - Gender and racial issues that affect delivery of care are we prepared for the future changes? PMID- 12627698 TI - Assess ABCD to ascertain the level of cultural influence in pediatric dental families. PMID- 12627699 TI - Indirect pulp treatment of primary posterior teeth: a retrospective study. AB - PURPOSE: This study was performed to assess retrospectively the clinical and radiographic success of indirect pulp treatment (IPT) on primary posterior teeth, and to compare the influence of caries risk, skills of the operator, and restorative material on the success of IPT. METHODS: A retrospective review of records of patients treated with IPT in the pediatric dental clinic at The University of Michigan, School of Dentistry from July 1993 through July 1999 was completed in January 2000. Two hundred fifty-five records with IPT were reviewed, from which 132 patients met the inclusion criteria, with 187 primary posterior teeth treated with an IPT. The patients were followed clinically and radiographically for a time ranging between 2 weeks to 73 months. Data were analyzed using survival analysis methods. RESULTS: The success of IPT was 95% (178/187 teeth), with only 9 failures. The 1-year probability of survival of each tooth was estimated to be 96% using an exponential survival model. The use of a base over a calcium hydroxide liner significantly increased the success rate of IPT (P = .0095). The use of a stainless steel crown (SSC) after an IPT was significantly more successful than the use of an amalgam (P=.026). IPT performed on primary first molars failed more frequently than on second primary molars (P = .045). There was no significant difference between maxillary and mandibular primary molars. CONCLUSIONS: Indirect pulp treatment is a successful technique and should be considered as an alternative pulp therapy procedure in deeply carious primary posterior teeth. The use of a base over the liner in addition to a SSC dramatically increases the success of an IPT. PMID- 12627700 TI - Caries experience in children with various genetic sensitivity levels to the bitter taste of 6-n-propylthiouracil (PROP): a pilot study. AB - PURPOSE: The objective of this pilot study was to determine the prevalence of coronal dental caries among children with different genetic sensitivity levels of taste, as determined by 6-n-propylthiouracil (PROP). METHODS: Coronal caries and restorations in permanent and primary dentition were evaluated in 150 healthy school-aged children aged 6 to 12 years. A filter paper containing 6-n propylthiouracil was used to determine each subject's genetic ability to taste bitter and sweet substances. Supertasters perceived stronger tastes from a variety of bitter and sweet substances than both medium tasters and nontasters. The data were analyzed by ANOVA with Duncan's multiple range test, Mantel Haenszel chi-square, multiple linear regression analyses, and Pearson's coefficient of correlation. RESULTS: The nontasters had more mean decayed, missing, and filled surfaces (dfs/DMFS) than tasters. The values of mean decayed and filled surfaces of primary dentition and mean decayed, missing, and filled surfaces of permanent dentition (dfs/DMFS) and mean decayed surfaces of primary dentition and permanent dentition (ds/DS) were significantly higher in nontasters than in medium tasters, and in medium tasters compared with supertasters. After adjusting for missing teeth, the data were expressed as a percentage of the available surfaces, and the significant differences in dfs/DMFS and ds/DS persisted (r = -0.49, P < .001 and r = -0.51, P < .0001 respectively). CONCLUSIONS: After all associated factors were controlled, taste was the only independent variable significantly related to overall caries experience. The results of this study suggested an increased prevalence of overall caries experience in nontaster children. PMID- 12627701 TI - Surface antibacterial properties of fissure sealants. AB - PURPOSE: Sealants form a physical barrier between the oral environment and deep fissures that contribute to caries prevention. It is postulated that sealants possessing antibacterial properties are advantageous. The purpose of this study was to evaluate the antibacterial properties of four pit and fissure sealants using direct contact test (DCT) and agar diffusion test (ADT). METHODS: For the DCT, 8 samples of Helioseal F (Vivadent), Ultraseal XT (Ultadent-Weldent), Conseal F (SDI), and Dyract Seal (Dentsply) were placed on the sidewalls of wells of a 96-microtiter plate. After polymerization, freshly grown Streptococcus mutans cells (circa 1 x 10(6)) were placed on the surface of each sample for 1 hour at 37 degrees C. Fresh media was then added to each well and bacterial growth was followed for 16 hours by temperature-controlled spectrophotometer. Similarly prepared samples were aged in phosphate buffered saline for 14 and 30 days and the DCT was repeated. The ADT was performed by placing samples in uniform wells punched in agar plates. RESULTS: Freshly polymerized samples in the DCT, Dyract Seal and Ultraseal XT possessed prominent antibacterial properties. Dyract Seal also demonstrated the most potent antibacterial properties, which lasted 14 days but faded within 30 days. In ADT, the halo in the bacterial lawn was measured after 48 hours, and only Dyract Seal demonstrated an inhibition zone. CONCLUSIONS: The compomer-based sealant Dyract Seal possessed the most potent and longest lasting antibacterial activity. PMID- 12627702 TI - Bond strength to primary tooth dentin following disinfection with a chlorhexidine solution: an in vitro study. AB - PURPOSE: The aim of this study was to determine the effect on shear bond strength of chlorhexidine used as a cavity disinfectant on primary tooth dentin. METHODS: Thirty specimens were randomly divided into 3 groups (N = 10) and treated as follows: (1) in Group I, the dentin was acid etched with a 37% phosphoric acid for 15 seconds, washed and dried; (2) in Group II, a 2% commercial chlorhexidine solution (Cav Clean) was applied for 40 seconds, washed and dried following acid etching for 15 seconds; and (3) in Group III, dentin was treated with a 37% phosphoric acid gel containing 2% digluconate of chlorhexidine (Cond AC) for 15 seconds. In all specimens, the adhesive Single Bond (3M) was applied, and composite cylinders (Filtek Z 250) were built. The specimens were sheared with a universal testing machine (Instron) running at a crosshead speed of 0.5 mm/min, and the results calculated in MPa. The specimens were also observed to record the failure mode. RESULTS: ANOVA analysis revealed that the shear bond strength of Group II (17.99+1.15 MPa) was significantly lower than Group I (19.88+1.02 MPa) and Group III (19.57+1.02 MPa). After debonding, 63% of the specimens presented cohesive failure of the material, 24% adhesive failure and 10% cohesive failure of dentin. CONCLUSIONS: The commercial cavity disinfectant containing 2% chlorhexidine had an adverse effect on Single Bond and produced significantly lower shear bond strength than the etch gel with chlorhexidine and the acid etch alone. PMID- 12627704 TI - Self-injurious behavior as a challenge for the dental practice: a case report. AB - Patients who present self-injurious behavior (SIB) can be considered a treatment challenge in the dental office or hospital setting. Oral structures can play various roles in the process of self injury. In this paper, the authors report a case of SIB in the form of cheek-biting on an adolescent with a history of hydrocephalus and developmental delay, which was successfully treated with a modified standard maxillary orthodontic retainer. Early detection and intervention in SIB cases in patients with developmental disabilities influence the successful outcome of the therapeutic interventions, enhancing the patient's quality of life. PMID- 12627703 TI - Attitudes of pediatric dentists towards tobacco intervention for children and adolescents: a pilot survey. AB - PURPOSE: The purpose of this study was to obtain pilot information concerning the attitudes of pediatric dentists regarding their role in tobacco intervention for children and adolescent patients. METHODS: A 1-page survey, which was adapted and pretested from a previous, comprehensive survey, was distributed to all registrants at the 2001 American Academy of Pediatric Dentistry Annual Meeting in Atlanta, Ga. Descriptive and exploratory multivariable techniques were used to analyze the data. RESULTS: Of the 173 respondents, only 18% had ever received training in tobacco cessation counseling. Although most dentists agreed that they should play a role in tobacco intervention with their young patients, the majority was not comfortable with this task. Pediatric dentists with tobacco counseling training were more likely to accept their role in tobacco prevention and cessation efforts and were more confident in their ability to do so than were their counterparts. Pediatric dentists consider tobacco cessation an important role in their practices, but do not consider themselves well prepared to act on this responsibility. CONCLUSIONS: The results of this pilot survey strongly suggest that a similar, national, comprehensive study should be conducted among pediatric dentists. PMID- 12627706 TI - Restoring coronal contours of retained infraoccluded primary second molars using bonded resin-based composite. AB - The purpose of this case report is to describe a step-by-step method of restoring function (and esthetics) to infraoccluded second primary molars using a crown form crown, in a case of bilateral congenital missing premolars. This technique restores form and function, with a minimal reduction to the tooth. It also conserves tooth material and permits good bonding of the resin-based composite to the enamel. The crown-form resin-based composite restoration is an easy and inexpensive solution in cases of infraoccluded teeth. Yet, patients must be warned about the possible failure of the crown and the need of periodic follow-up appointments. PMID- 12627705 TI - Simplified primary incisor proximal restoration. AB - Primary anterior teeth with proximal caries lesions can be restored with resin based composite using a simplified bonding technique. This report describes Class III restoration of primary incisors in a preschooler, using a self-etching adhesive system and a modified-bonding procedure. PMID- 12627707 TI - Telomerase: a potential diagnostic and therapeutic tool in canine oncology. AB - In recent years there has been considerable interest in telomerase as a target for therapeutic intervention in oncology. This largely stems from the vast number of studies that have demonstrated expression and activity of the enzyme telomerase in the majority of human cancer tissues with little or no activity detectable in normal somatic tissues. These studies have led to an interest in the role of telomerase in cancers associated with domesticated species, in particular tumors that affect dogs. This article reviews the biology of telomerase and the biological significance of telomerase activity in canine tumors and discusses the clinical implications of telomerase expression in canine cancers with regard to therapeutics and diagnostics. PMID- 12627708 TI - Pancreatic injury in equine acute abdomen evaluated by plasma trypsin activity and histopathology of pancreatic tissue. AB - In cases of equine acute abdominal disease, where pancreatic damage is suspected, pancreatic damage can be assessed by measuring increased trypsin activity in the plasma of horses suffering intestinal obstruction and severe shock. The pancreas is particularly vulnerable to splanchnic hypoperfusion because it is a highly active tissue. In this study, 10 horses undergoing abdominal surgery for intestinal obstruction were assayed for trypsin activity on admission and, because of extensive intestinal lesions that were not amenable to surgery, euthanasia was selected; the pancreas was removed before euthanasia. Trypsin activity in the plasma of these horses was significantly higher than in healthy horses (196 ng/ml +/- 128.2 versus 28.5 ng/ml +/- 19.2; P = 0.0026). Light and transmission electron microscopy revealed slight to severe lesions of vacuolar degeneration, a few zymogen granules, dilation of the endoplasmic reticulum, and swelling of mitochondria in the exocrine pancreas. The activation of an inflammatory cascade occurring during strangulating intestinal obstruction could increase pancreatic anoxic lesions caused by severe shock and hypoperfusion in the horse. Further studies will show the significance of pancreatic lesions and the ensuing damage in equine acute intestinal obstruction and shock. PMID- 12627709 TI - Varied pathogenicity of a Hong Kong-origin H5N1 avian influenza virus in four passerine species and budgerigars. AB - This investigation assessed the ability of the zoonotic A/chicken/Hong Kong/220/97 (chicken/Hong Kong) (H5N1) highly pathogenic avian influenza virus to infect and cause disease in zebra finches (Taeniopygia guttata), house finches (Carpodacus mexicanus), house sparrows (Passer domesticus), European starlings (Sternus vulgaris), and budgerigars (Melopsittacus undulatus) after intranasal administration. Zebra finches were the most severely affected of the five species, demonstrating anorexia, depression, and 100% mortality within 5 days of inoculation. Gross lesions in this species were absent or only mild. But histologic lesions and the corresponding viral antigen were observed in multiple organs, especially in the nasal cavity, brain, pancreas, spleen, adrenal glands, and ovary. Significant morbidity and mortality also were observed in both house finches and budgerigars. Affected birds of these two species demonstrated anorexia, depression, and neurologic signs and typically were moribund or dead within 2 days of the onset of clinical signs. Gross lesions were mild or absent in house finches and budgerigars. Histologically, the brain and pancreas were the most consistently and severely affected organs in house finches. The brain was the most affected organ in budgerigars. Unlike these three species, house sparrows suffered only mild transient depression, had no mortality, and lacked gross lesions. Viral antigen and microscopic lesions were observed only in the heart and testicle of a minority of birds of this species. Starlings demonstrated neither clinical disease nor mortality and lacked gross and histologic lesions. Viral antigen was not observed in any of the collected tissues from starlings. These results indicate that there is significant variation in the pathogenicity of the chicken/Hong Kong virus for different species of birds, including species within the same order. In addition, neurotropism is a recurrent feature among birds that eventually succumb to infection. PMID- 12627710 TI - Expression of cyclooxygenase-2 in swine naturally infected with Actinobacillus pleuropneumoniae. AB - Cyclooxygenase-2 (COX-2) was detected and localized in 15 pigs with naturally occurring pleuropneumonia using a 437-base pair digoxigenin-labeled cDNA probe in an in situ hybridization protocol. Histopathologic changes in the acute stage were characterized by coagulative necrosis of lung parenchyma, hemorrhage, vascular thrombosis, edema, fibrin deposition, and infiltration of lung parenchyma by neutrophils and alveolar macrophages in nine pigs. In chronic lesions, a thick layer of granulation tissue surrounded foci of pulmonary necrosis in six pigs. All 15 pigs infected with Actinobacillus pleuropneumoniae, confirmed by bacterial isolation, had distinct positive hybridization signals for COX-2 in bronchial, bronchiolar epithelial cells, alveolar macrophages, neutrophils, and type I pneumocytes. COX-2 expression was detected primarily in neutrophils from pigs with acute lesions and primarily in alveolar macrophages from pigs with chronic lesions. The results suggest that a prostanoid product of COX-2 is an important component of the inflammatory response to acute and chronic A. pleuropneumoniae infection. PMID- 12627711 TI - Diagnosis of canine lymphoid neoplasia using clonal rearrangements of antigen receptor genes. AB - Although the diagnosis of canine leukemia and lymphoma in advanced stages is usually uncomplicated, some presentations of the disease can be a diagnostic challenge. In certain situations, lymphoma and leukemia can be difficult to distinguish from a benign reactive proliferation of lymphocytes. Because clonality is the hallmark of malignancy, we have developed an assay that uses the polymerase chain reaction to amplify the variable regions of immunoglobulin genes and T-cell receptor genes to detect the presence of a clonal lymphocyte population. The assay detected clonally rearranged antigen receptor genes in 91% of the 77 dogs with lymphoid malignancy. Of the 24 dogs tested, that were either healthy or had clearly defined conditions not related to lymphoid malignancy, a clonally rearranged antigen receptor gene was found in one (a dog with Ehrlichia canis infection). Gene rearrangement was appropriate for the immunophenotype (immunoglobulin gene rearrangement in B-cell leukemias and T-cell receptor gene rearrangement in T-cell leukemias). Dilution analysis showed that the clonal rearrangement could be detected when 0.1-10% of the DNA was derived from neoplastic cells, depending on the source tissue. Potential applications of this assay include the diagnosis of lymphoma or leukemia in biopsy samples, cavity fluids, fine needle aspirates, bone marrow and peripheral blood; the determination of lineage (B or T cell); staging of lymphoma; and detection of residual disease after chemotherapy. PMID- 12627712 TI - Gastrointestinal stromal tumors and leiomyomas in the dog: a histopathologic, immunohistochemical, and molecular genetic study of 50 cases. AB - Fifty canine gastrointestinal (GI) mesenchymal tumors were examined to determine the occurrence of leiomyomas (LM) and GI stromal tumors and to compare their clinicopathologic features. Twenty-one tumors (42%) were histologically reclassified as gastrointestinal stromal tumors (GISTs) and 29 tumors (58%) as LMs on the basis of their histologic similarity with homologous human tumors. The GISTs occurred equally in males and females, with a mean age of 11 years (range 5 14 years). Five GISTs (24%) were associated with clinical signs and six (29%) had metastasis in liver or abdominal cavity. The GISTs occurred in large intestine (10, 48%), small bowel (six, 29%), stomach (four, 19%), and mesentery of small intestine (one, 5%). Histologically, they were highly cellular spindle, or less commonly epithelioid tumors with mitotic rates ranging from 0 to 19 per 10 HPF. Eleven tumors (52%) were positive for CD117 (KIT); seven (33%) were positive for smooth muscle actin but none for desmin and S-100 protein. Sequences of KIT exon 11, often mutated in human GISTs, were evaluated from four GISTs. Deletion of Try556-Lys557 coexisting with duplication of Gln555 in one case of GIST and T to C transition resulting in substitution of Pro for Leu575 in another were identified. The LMs occurred predominantly in males (82%) with a mean age of 11 years (range 8-17 years). Nine tumors (31%) had associated clinical signs. They occurred in the stomach (22, 76%), esophagus (four, 14%), and intestines (three, 10%); all were paucicellular, had no mitoses, and were composed of mature smooth muscle cells. Twenty-eight (97%) were positive for smooth muscle actin and 18 (62%) for desmin but none for CD117 and S-100. Both GISTs and true LMs occur in the GI tract of dogs. Both tumors have distinctive pathologic features. PMID- 12627713 TI - Immunohistochemical detection of uroplakin III, cytokeratin 7, and cytokeratin 20 in canine urothelial tumors. AB - Immunohistochemistry for uroplakin III (UP III), cytokeratin 7 (CK 7), and cytokeratin 20 (CK 20) using commercially available antibodies was done in normal canine urinary bladder and 72 canine urinary bladder tumors that had been fixed in formalin and embedded in paraffin. Prolonged fixation (3-28 days) did not significantly alter the immunostaining for UP III. There was moderate reduction in the intensity for CK 7 and CK 20 after 1 week of fixation. UP III was detected in superficial (umbrella) cells and some intermediate cells of the normal urinary bladder, 7 of 7 transitional cell papillomas (TCPs), 50 of 55 transitional cell carcinomas (TCCs), and 4 of 5 metastatic TCCs. Staining was typically outlined in the plasma membrane, but diffuse or focal cytoplasmic staining was also observed. Intracytoplasmic lumina were usually positive for UP III. One squamous cell carcinoma of the bladder, 4 nonepithelial bladder tumors, and 285 nonurothelial tumors from different nonurinary locations were negative for UP III. CK 7 was detected in 7 of 7 TCPs, 53 of 54 TCCs, and 5 of 5 metastatic TCCs. The staining for CK 7 was diffuse cytoplasmic. CK 20 was detected in 1 of 7 TCPs, 37 of 54 TCCs, and 1 of 5 metastatic TCCs. The staining with CK 20 was cytoplasmic and weaker than with antibodies to UP III or CK 7. There was concurrent expression of UP III, CK 7, and CK 20 in 36 of 54 TCCs. UP III is a specific and sensitive marker for canine transitional epithelial (urothelial) neoplasms, detecting 91% of TCCs. Negative results may be observed with anaplastic tumors. PMID- 12627714 TI - A familial peripheral neuropathy and glomerulopathy in Gelbvieh calves. AB - Nine Gelbvieh calves originating in four herds and clinically presenting with rear limb ataxia/paresis had histopathologically confirmed peripheral neuropathy and a proliferative glomerulopathy. Degenerative lesions were severe in peripheral nerves, dorsal and ventral spinal nerve roots, and less marked in dorsal fasciculi of the spinal cord. Cell bodies of spinal ganglia were minimally diseased; ventral horn neurons occasionally had central chromatolysis and nuclear displacement. Glomerular lesions ranged from mild mesangial hypercellularity to glomerulosclerosis. Pedigree analysis of affected animals from one herd indicated a strong familial relationship and probable hereditary basis for the syndrome. PMID- 12627715 TI - Pathology of AA amyloidosis in domestic sheep and goats. AB - We describe the main pathologic changes in small ruminants affected by AA amyloidosis, together with the partial sequence of the protein involved. Twenty one sheep and one goat were selected for presenting macroscopic kidney lesions compatible with systemic amyloidosis. Available tissue samples were studied by histologic, immunopathologic, and ultrastructural means. Renal lesions were characterized grossly by pale cortical surfaces with scattered, miliary, whitish yellow foci and on cut cortical surfaces by straight, whitish-yellow striations. Gangrenous pneumonia was observed in 16 out of 21 affected sheep (76.2%), although other chronic inflammations were also observed. Amyloid was detected in all grossly affected kidneys using Congo red staining, lesions being most remarkable in glomeruli, affecting 95.5% of animals studied. Congophilic deposits were also observed in intertubular interstitium (68.2%) and medulla (57.1%). All amyloid-affected animals presented proximal convoluted tubule lesions, mostly characterized by an increase in diameter and by hyaline granular degeneration that were responsible for the macroscopic appearance of the kidney. Histologically, amyloid was also seen in blood vessels, spleen, liver, lymph nodes, gastrointestinal tract, and adrenal glands. All amyloid deposits demonstrated greenish-yellow birefringence with polarized light, and the antisera prepared against goat amyloid extracts specifically reacted with birefringent congophilic deposits of both sheep and goats. Ultrastructurally, these deposits were formed by masses of straight, nonbranching fibrils located predominantly in the basement membranes of glomerular capillaries and in the mesangium. Partial sequence of the protein in sheep and goats indicated a high degree of homology with the previously reported sequence of sheep Serum Amyloid A. PMID- 12627716 TI - Preliminary observations on the experimental transmission of scrapie to elk (Cervus elaphus nelsoni) by intracerebral inoculation. AB - To determine the transmissibility of scrapie to Rocky Mountain elk (Cervus elaphus nelsoni), six elk calves were inoculated intracerebrally with brain suspension from sheep naturally affected with scrapie. One elk developed a brain abscess and was euthanatized at 7 weeks postinoculation (PI), and two others died at 6 and 15 months PI because of physical injuries. At 25 and 35 months PI, two other elk died after brief terminal neurologic episodes. Necropsy of these revealed moderate weight loss but no other gross lesions. Microscopically, characteristic lesions of spongiform encephalopathy were seen throughout the brains and the spinal cords, and in both cases these tissues were positive for PrP(res) by immunohistochemistry. Brains of both animals were positive for PrP(res) by western blot and for scrapie-associated fibrils (SAFs) by negative stain electron microscopy. PrP(res) and SAFs were not detected in the three elk that died or were euthanatized because of coincidental causes. Over 3.5 years after initiation of this experiment, the one remaining inoculated elk and two uninoculated (control) elk are alive and apparently healthy. These preliminary findings demonstrate that 1) sheep scrapie agent can be transmitted to elk by intracerebral inoculation; 2) the infection can result in severe, widely distributed spongiform change and accumulations of PrP(res) in the central nervous system (CNS); and 3) based on the examination of a limited number of CNS sections from two cases, this condition cannot be distinguished from chronic wasting disease with currently available diagnostic techniques. PMID- 12627717 TI - Invasive ductal carcinoma of the mammary gland in a mare. AB - A 21-year-old thoroughbred mare had a 35 x 14 x 10 cm mass involving the mammary gland. Metastases were found in the kidneys, lungs, skeletal muscles, and regional lymph nodes. Histopathologic examination of the tumor revealed a ductal solid carcinoma with extensive intraductal and intralobular involvement and focal infiltration of the adjacent stroma. The intralobular neoplasms were divided into irregularly shaped islands and sheets of polygonal and spindle-shaped epithelial cells by thick or thin fibrous connective tissue bundles. The neoplastic cells had a small or moderate amount of cytoplasm that stained faintly with eosin and round or oval hyperchromatic nuclei. Immunohistochemically, the neoplastic cells were strongly positive for Lu-5, weakly positive for AE1/AE3, vimentin, and glial fibrillary acidic protein, and negative for cytokeratin 8, cytokeratin 14, alpha smooth muscle actin, calponin, and S100. The neoplasm was diagnosed as an invasive ductal carcinoma of the mammary gland with multiple metastases. PMID- 12627718 TI - Intramedullary hemangioblastoma in a dog. AB - A 6-year-old male Pointer dog was presented with a 4-week history of progressive hind-limb stiffness. Magnetic resonance imaging demonstrated a focal intramedullary lesion at T1 level with a pattern of ring contrast enhancement. At necropsy, a circumscribed intramedullary reddish-gray tumor was observed. Microscopically, the tumor was composed of thin-walled capillaries lined by endothelial cells and separated by pleomorphic cells (stromal cells) with a moderate degree of anisokaryosis. Immunohistochemically, the endothelial cells were positive for factor VIII-related antigen and the stromal cells were positive for neuron-specific enolase and vimentin. GFAP-positive astrocytes were occasionally observed within the tumor. Both endothelial and stromal cells were negative for synaptophysin, S-100 protein, pankeratin, smooth muscle actin, CD34, CD68, alpha1-antichymotrypsin, and lysozyme. The tumor showed considerable morphologic and immunohistochemical similarities with human hemangioblastoma, and hence the inclusion of this tumor type within the primary neoplasms of the canine central nervous system is suggested. PMID- 12627719 TI - Vasculitis in a dog resembling isolated angiitis of the central nervous system in humans. AB - An 1-year-old dog succumbed to a seizure disorder of 18 days duration. At necropsy, an area of hemorrhage and discolored parenchyma was identified in the left pyriform lobe of the brain. Microscopic examination revealed a localized, necrotizing vasculitis with associated cerebral necrosis. Vasculitis was not present in other organs. This presentation is consistent with isolated central nervous system (CNS) angiitis, a rare form of vasculitis in humans. PMID- 12627720 TI - Gliomatosis cerebri in six dogs. AB - Gliomatosis cerebri is a well-recognized entity in human medicine characterized by unusually widespread infiltration of the neuraxis by neoplastic glial cells with relative preservation of brain architecture. This report describes the pathologic features of the disease in six dogs. The dogs ranged from 3 to 9 years of age (mean 6.1 years) without evidence of breed predilection; five of the six dogs were neutered or intact males. The clinical findings were mixed (including depression, circling, cranial nerve deficits), reflecting the diffuse nature of the disease. Histologically, there was remarkably diffuse infiltration of the white and gray matter of the brain by small numbers of elongated neoplastic cells. Areas of greater cellularity formed grossly visible lesions in four cases. Anisocytosis and pleomorphism were greater in areas of higher cellularity. Other features of tumor growth included subpial accumulation, neuronal satellitosis, perivascular cuffing, and tropism for cranial nerve and brain stem nuclei. Neoplastic cells were negative on immunohistochemical stains for glial fibrillary acid protein (GFAP) and leukocyte markers, reflecting the uncertain histogenesis of these unusual neoplasms. PMID- 12627721 TI - Camelid mucoutaneous fibropapillomas: clinicopathologic findings and association with papillomavirus. AB - Five camelid mucocutaneous fibropapillomas with histologic features similar to equine sarcoids were diagnosed. They were characterized by a dermal fibroblastic proliferation and overlying, often ulcerated hyperplastic epidermis with thin rete pegs extending down into the dermis. Two of the tumors came from llamas and three from alpacas. Four of the animals were 6-year-old females. The fifth was a 6-year-old castrated male. The fibropapillomas were located on the nose, lip, and cheeks. One of the llama tumors waxed and waned before surgery and recurred and spread after surgery. None of the other tumors recurred. All five tumors were positive for papillomavirus (PV) DNA by polymerase chain reaction testing. Nucleotide sequence analysis of the PCR product from one of the llama fibropapillomas confirmed a unique PV. This report provides the microscopic and clinical features of fibropapillomas in camelids as well as evidence for a PV etiology. PMID- 12627722 TI - Localization of classical swine fever virus from chronically infected pigs by in situ hybridization and immunohistochemistry. AB - Classical swine fever (CSF) virus (CSFV) nucleic acid and antigen were detected in 15 pigs with naturally occurring chronic CSF by in situ hybridization and immunohistochemistry. The most consistent and prominent microscopic lesions were perivascular mononuclear cell infiltration and gliosis in the central nervous system of pigs with chronic CSF. Positive cells typically exhibited a dark brown (in situ hybridization) or red (immunohistochemistry) reaction product in the cytoplasm without background staining. A positive signal for both in situ hybridization and immunohistochemistry was detected in mononuclear cells and lymphocytes of lymphoid tissues. Viral nucleic acid was detected in some tissue sections in the absence of viral antigen. The in situ hybridization technique developed in this study was useful for the detection of CSFV RNA in tissues taken from chronically infected pigs and may be a valuable technique for studying the pathogenesis of chronic CSFV infection. PMID- 12627723 TI - Hepatic biliary adenofibroma: a hitherto unrecognized tumor in equines. Report of a case. AB - Hepatic biliary cystadenoma is a well-delineated neoplasm in some domestic animals, especially in cats, but it has not been reported in equines. We report on a case of hepatic biliary tumor, incidentally found in a 10-year-old horse, with gross and microscopic features similar to those observed in biliary adenofibroma of humans. The tumor presented as a solid mass measuring 16 cm in diameter and histologically was composed of complex tubulocystic biliary components embedded in an abundant fibrotic stroma. We regarded this tumor as a morphological variant of biliary cystadenoma of domestic animals. Differential diagnoses from other hepatic biliary tumor-like and tumor lesions are provided. PMID- 12627724 TI - B-cell leukemia in a sheep. AB - Necropsy examination was performed on an 8.5-year-old Finnish ewe euthanatized because of progressive respiratory distress, cachexia, and weakness. Significant postmortem findings included a diffusely enlarged, dark-red friable liver, mild splenomegaly, and mild mesenteric lymphadenopathy. Examination of multiple tissue sections revealed intravascular atypical mononuclear cells in all tissues examined, with a leukemic pattern of infiltration of mesenteric lymph nodes, liver, lung, and spleen. Neoplastic cells were positive for CD79a and negative for CD-3, BLA.36, and CD68 leukocytic markers. The final diagnosis was B-cell leukemia. PMID- 12627725 TI - Possible contributing factors of sleep apnea syndrome to resistant hypertension. PMID- 12627726 TI - Polysomnographic sleep state from wrist activity. PMID- 12627727 TI - Sympathetic over activity in the etiology of hypertension of obstructive sleep apnea. AB - Beginning with modest clinical observations in 1984, a picture has evolved suggesting that sympathetic nervous system over activity may be responsible in part for the elevated blood pressure seen in obstructive sleep apnea patients. Early studies of urinary and plasma catecholamines indirectly suggested sympathetic over activity carried to daytime, non-apneic conditions. Later intra neuronal recordings of muscle sympathetic nerve activity directly demonstrated both acute and diumal (non-apneic) sympathetic over activity. Most importantly, diurnal sympathetic over activity has been shown to diminish with adequate treatment of apnea using nasal CPAP. Norepinephrine and angiotensin II are both released with increased peripheral sympathetic activity and are parallel vascular growth-promoting factors. Thus, one would expect alterations in vascular structure and function in a state of chronic sympathetic over activity. While changes in peripheral vascular structure have not been demonstrated in hypertension of sleep apnea, changes in peripheral vascular responsiveness have. There is reduced response to acetylcholine and isoproterenol vasodilation, and to norepinephrine and angiotensin vasoconstriction in humans with sleep apnea. Some of these vascular reactivity changes are shown to reversed with chronic nasal CPAP treatment. Finally, complimentary to the above evidence in humans, there is indirect evidence of sympathetic over activity as well as differences in vascular reactivity in intermittent hypoxia challenged rats. We have made significant strides in the past 15-20 years towards understanding systemic hypertension related to sleep apnea, especially the role of the sympathetic nervous system. Future research will need to look at exact mechanism of sympathetic nervous system over activity, particularly how central nervous system pathways may undergo facilitation, leading to daytime over activity. Furthermore, the mechanisms of sustained hypertension in sleep apnea patients is almost certainly of multiple etiologies. There is no marker for separating sleep apnea patients with hypertension derived solely from intermittent hypoxia from other secondary causes. Perhaps endothelial cell molecular markers could help to identify patients at risk for cardiovascular change associated with snoring and apnea, as well to guide treatment. Finally, studies demonstrating microvascular changes in blood vessels are extremely difficult to do, but promise to yield important knowledge about cellular mechanisms and results of long-term treatment of sleep apnea on cardiovascular disease. PMID- 12627729 TI - A 12-month, open-label, multicenter extension trial of orally administered sodium oxybate for the treatment of narcolepsy. AB - OBJECTIVES: To evaluate the long-term safety and efficacy of nightly sodium oxybate for the treatment of narcolepsy. DESIGN: A multicenter, 12-month, open label trial. PARTICIPANTS: 118 narcolepsy patients previously enrolled in a 4 week double-blind sodium oxybate trial. INTERVENTIONS: Patients were administered 6 g sodium oxybate nightly, taken in equally divided doses at bedtime and 2.5 to 4 hours later. The study protocol permitted the dose to be increased or decreased in 1.5-g increments at 2-week intervals based on efficacy response or adverse experiences but staying within the range of 3 to 9 g nightly. MEASURES: Narcolepsy symptoms and adverse events were recorded in daily diaries. Safety measures included physical and laboratory examinations repeated at 6 and 12 months. The primary efficacy measure was the change in weekly cataplexy attacks from baseline. Secondary measures included daytime sleepiness using the Epworth Sleepiness Scale (ESS), inadvertent naps/sleep attacks, nighttime awakenings, and the overall change in disease severity as rated by the investigators (Clinical Global Impression of Change; CGI-c). RESULTS: Sodium oxybate, in doses of 3 to 9 g nightly, produced overall improvements in narcolepsy symptoms, which were significant at 4 weeks and maximal after 8 weeks. Reported improvements included a significant decrease in frequency of cataplexy attacks (p < 0.001); diminished daytime sleepiness (p < 0.001); and patient descriptions of nocturnal sleep quality, level of alertness, and ability to concentrate (for each p < 0.001). Adverse events were generally mild and patients showed no evidence of tolerance. CONCLUSION: Sodium oxybate is an effective and well-tolerated treatment for narcolepsy. PMID- 12627728 TI - Hypocretinergic neurons are primarily involved in activation of the somatomotor system. AB - The hypocretinergic system has been implicated in the generation and/or maintenance of wakefulness. Our results challenge this hypothesis. Utilizing cats as an animal model and immunocytochemical procedures for the simultaneous detection of hypocretin and Fos, we determined that hypocretinergic neurons are activated during wakefulness but only when somatomotor activity is present. These neurons are not activated during alert or quiet wakefulness in the absence of motor activity or during quiet sleep. We conclude that the hypocretinergic system is not responsible for the generation and/or maintenance of wakefulness, per se; on the contrary, we suggest that hypocretinergic neurons are primarily involved in motor functions irrespective of the animal's behavioral state. PMID- 12627730 TI - Attention deficits in patients with narcolepsy. AB - STUDY OBJECTIVES: Although attention problems are presumably responsible for a wide variety of difficulties patients with narcolepsy experience in everyday life, empirical investigation of this issue is scarce. Therefore, we conducted a systematic investigation of different aspects of attention and verbal memory in patients with narcolepsy. DESIGN: Control-group design with comparison of performance in four attention tests--measuring phasic alertness, focused attention, divided attention, and flexible attention--and one verbal memory test. PARTICIPANTS: 19 patients with narcolepsy (NG) and 20 healthy controls (CG) MEASUREMENTS AND RESULTS: The NG showed no deficits in phasic alertness, focused attention, and verbal memory. However, specific deficits occurred in divided and flexible attention. Furthermore, the NG had generally slower and more variable reaction times in all attention tasks. CONCLUSIONS: Our results contradict the hypothesis that attentional impairments in narcolepsy are merely a result of a temporal disturbance of information processing, i.e., deficits can be explained by slowness and variability of performance alone. Rather, deficits in attentional capacity and attentional control also seem to play an important role. Thus, in addition to impairment in the vigilance attention network, results indicate impairment in the executive attention network in patients with narcolepsy. PMID- 12627731 TI - Voxel-based morphometry in hypocretin-deficient narcolepsy. AB - STUDY OBJECTIVES: Recent studies suggest that narcolepsy is caused by degeneration of hypocretin (orexin) producing neurons. To find evidence for this hypothesis, we aimed to detect structural changes in the hypothalamus and/or hypocretin projection areas of patients with narcolepsy. DESIGN: We used voxel based morphometry (VBM), an unbiased MRI morphometric method with a high sensitivity for subtle changes in gray and white matter volumes. SETTING: Image acquisition was carried out in the department of Radiology at Leiden University Medical Center; image post-processing was performed in the Wellcome Department of Cognitive Neurology, London. PARTICIPANTS: Fifteen narcoleptic patients were studied, all having cataplexy and typical findings on Multiple Sleep Latency Testing. All patients were HLA-DQB1*0602 positive and hypocretin-1 deficient. The control group consisted of 15 age and sex matched healthy subjects. MEASUREMENTS AND RESULTS: We found no differences in global gray or white matter volumes between patients and controls. Furthermore, regional gray or white matter volumes in the hypothalamus and hypocretin projection areas did not differ between patients and controls. CONCLUSIONS: VBM failed to show structural changes in the brains of patients with narcolepsy. This suggests that narcolepsy either is associated with microscopic changes undetectable by VBM or that functional abnormalities of hypocretin neurons are not associated with structural correlates. PMID- 12627732 TI - Clinicians' use of the International Classification of Sleep Disorders: results of a national survey. AB - OBJECTIVE: To determine clinicians' patterns of use of the International Classification of Sleep Disorders (ICSD) and their ratings of its utility and organization. METHODS: A telephone survey was conducted in one clinician practicing at each of 206 sleep centers accredited by the American Academy of Sleep Medicine (AASM). Descriptive analyses were performed, and ratings for various sleep classifications were compared with ANOVA and post- hoc tests. RESULTS: Clinicians at 230/251 (92%) of contacted centers agreed to participate, and data from 206/251 (82%) were actually collected. The ICSD was used by 91.7% of clinicians for clinical decision making but by lower percentages for billing or medical record keeping. Features of the ICSD pertaining to clinical descriptions and differential diagnosis were rated as most useful. The Association of Sleep Disorders Centers (ASDC) classification was rated more highly than ICSD in organization and ease of use. The ICSD and ASDC systems were rated more highly than other sleep disorders classifications in terms of organization, "fit" to patients, and ease of use. CONCLUSIONS: The ICSD is used widely by clinicians, primarily for diagnostic purposes, and is preferred over more general sleep disorders classification systems. However, more favorable ratings for the ASDC compared to ICSD indicate the value clinicians place on easy to-use, symptom-oriented classifications. PMID- 12627733 TI - Hypoglycemia counterregulation during sleep. AB - STUDY OBJECTIVES: In insulin-treated patients with diabetes, episodes of severe hypoglycemia often occur during sleep, which might reflect an altered counterregulation and reduced awareness. This study examined the influence of sleep on the counterregulatory response to hypoglycemia in healthy subjects. DESIGN: Subjects participated in two experimental conditions; statistical tests relied on within subject comparisons. SETTING: University hospital sleep laboratory. PARTICIPANTS: 15 healthy young men. INTERVENTIONS: Hypoglycemia (2.8 mmol/l) was induced for 45 min by insulin infusion once during sleep and once at the same time of night while being awake. MEASUREMENTS AND RESULTS: Counterregulatory hormone concentrations (epinephrine, norepinephrine, ACTH and cortisol) and sleep recordings were obtained. Differences in the hormonal responses to hypoglycemia between sleep and wake conditions remained non significant, indicating that sleep does not exert a primary influence on the strength of counterregulation. However, the glycemic threshold for the onset of counterregulation was significantly changed during sleep: The average onset threshold for epinephrine and norepinephrine counterregulation was 3.3 +/- 0.1 mmol/l for the wake condition and 2.7 +/- 0.1 mmol/l for the sleep condition (P < 0.001). A decrease in sleep depth coincided with the onset of the counterregulatory response, with most subjects showing signs of awakening. CONCLUSIONS: During sleep, the organism is less sensitive to hypoglycemia. Hypoglycemia per se has an awakening effect. PMID- 12627734 TI - Prevalence and predictors of nocturia in obstructive sleep apnea-hypopnea syndrome--a retrospective study. AB - OBJECTIVE: To determine the prevalence and predictors of pathological nocturia (PN) in patients with OSAHS. METHODS: Retrospective review of clinical and polysomnographic data obtained from patients evaluated at the Pulmonary Sleep Evaluation Center of the University of Pittsburgh Medical Center between November 1998 and September 1999. PN was defined as two or more urination events per night. OSAHS was defined as Apnea-Hypopnea Index (AHI) > or = 5. Group t-tests and Chi-square test were used to examine differences in subjects with and without PN. A multivariate approach utilizing logistic regression was performed to examine the relationships between polysomnographic variables [Arousal Index (ARI), AHI, Apnea-Hypopnea Time/Total Sleep Time x 100(%) (AHT/TST), % Total Sleep Time (TST)<90% Oxyhemoglobin Saturation (SPO2), nadir SPO2, Desaturation Event Frequency (average number of desaturations > 4% per hour sleep) (DEF)] and the presence of PN, while controlling for medical comorbidities. Data are expressed as mean + SD unless otherwise specified. Statistical significance was assessed at p < 0.05. RESULTS: STUDY POPULATION: n = 138 (50 females, 88 males), age: 49.7 +/- 12.3 years, BMI [Body Mass Index (kg/m2)]: 37.7 +/- 11.3. The overall prevalence of PN was 47.8%. The prevalence of PN was greater in females (60% vs. 40.9% in males). OSAHS patients with PN had a higher prevalence of peripheral edema, higher BMI, a greater %TST < 90% SPO2, lower nadir SPO2 and greater DEF. The logistic regression model indicated that age, ARI, AHI, AHT/TST, %TST < 90%, DEF were significant predictors of PN independent of BMI, neck circumference and medical comorbidities. CONCLUSIONS: Our data confirm that PN is common in OSAHS patients. The strongest predictors are age and selected polysomnographic variables reflecting OSAHS severity. PMID- 12627735 TI - Model-based assessment of autonomic control in obstructive sleep apnea syndrome. AB - STUDY OBJECTIVES: To quantitatively assess autonomic cardiovascular control in patients with obstructive sleep apnea syndrome (OSAS) using a mathematical model that relates changes in R-R interval (RRI) to respiration and changes in systolic blood pressure (SBP), and to compare the results obtained with conventional techniques. DESIGN: Respiration, RRI, and arterial blood pressure were monitored noninvasively in awake subjects in the supine and standing postures. A mathematical model was used to partition the fluctuations in RRI into a component ("RSA") correlated with respiration and a component ("baroreflex") correlated with fluctuations in SBP. SETTING: Sleep disorders laboratory in a hospital setting. PATIENTS OR PARTICIPANTS: 11 middle-aged male patients with untreated OSAS (apnea-hypopnea index = 75.9 +/- 11.1 (SE) events h-1) and 11 age-matched normal controls (10 males + 1 female). INTERVENTIONS: The subjects were monitored while breathing spontaneously in both supine and standing postures. Each subject also had to perform a battery of 5 standard autonomic stress tests (AST). MEASUREMENTS AND RESULTS: Four of the 5 ASTs did not indicate any difference between controls and OSAS subjects. There were also no differences in the indices derived from power spectral analysis of RRI and blood pressure variability; however, these spectral indices were sensitive to postural changes (orthostatic stress). Both RSA and baroreflex gains estimated from the model were substantially depressed in OSAS (P < 0.02) Changes in posture affected RSA gain but not baroreflex gain. The time-courses of the dynamics of these model components were not significantly different between subject groups. CONCLUSIONS: OSAS leads to abnormal parasympathetic and sympathetic control of heart rate, as reflected in depressed RSA and baroreflex gains. Model-based assessment was more sensitive in detecting abnormal autonomic function, compared to standard autonomic testing and power spectral analysis. The model-based approach represents a relatively simple and nonintrusive means of quantifying the key aspects of autonomic control in spontaneously breathing OSAS patients during wakefulness. PMID- 12627736 TI - Variation in symptoms of sleep-disordered breathing with race and ethnicity: the Sleep Heart Health Study. AB - STUDY OBJECTIVES: To examine the relation of sleep-related symptoms to race and ethnicity in a diverse sample of middle-aged and older men and women. DESIGN: Cross-sectional questionnaire survey. SETTING: In the initial phase of the Sleep Heart Health Study, men and women enrolled in participating epidemiologic cohort studies were surveyed. PARTICIPANTS: 13,194 men and women 40 years of age and older, including 11,517 non-Hispanic white, 648 black, 643 American Indian, 296 Hispanic, and 90 Asian-Pacific Islander. INTERVENTIONS: Not applicable. MEASUREMENTS AND RESULTS: After adjustment for BMI and other factors, frequent snoring was more common among Hispanic women (odds ratio (OR) = 2.25, 95% confidence interval (CI) = 1.48, 3.42) and black women (OR = 1.55, 95% Ci = 1.13, 2.13) than among non-Hispanic white women. Hispanic men were significantly more likely to report frequent snoring than non-Hispanic white men (OR = 2.30, 95% CI = 1.43, 3.69). Black, American Indian, and Asian men did not differ significantly from white men in snoring prevalence. American Indian women were significantly more likely to report breathing pauses during sleep than their white, non Hispanic counterparts (OR = 1.52, 95% CI 1.03, 2.24), although polysomnography data on a subset of the sample suggested that the association between this symptom reported on questionnaire and objective evidence of sleep-disordered breathing may be weaker among American Indians than among other groups. Mean Epworth Sleepiness Scale scores were slightly higher in black men and women than in their white, non-hispanic counterparts. CONCLUSIONS: Frequent snoring was more common among black and Hispanic women and Hispanic men than among their white non Hispanic counterparts, even after adjusting for BMI and other factors. Further research including polysomnography and objective measurements of sleepiness is needed to assess the physiologic and clinical significance of these findings. PMID- 12627737 TI - Further validation of actigraphy for sleep studies. AB - STUDY OBJECTIVES: Actigraphy is generally compared to polysomnography (PSG), which has been considered the gold standard for sleep studies. The objective of the present study was to evaluate the concordance between PSG and two previously proposed algorithms (Cole et al, 1992; Sadeh et al, 1994) to analyze actigraphic recordings. The minute-by-minute agreement rate was evaluated through calculation of sensitivity, specificity, and accuracy. Regarding the sleep parameters, the concordance was performed through the Bland and Altman technique. DESIGN: A night of adaptation to the sleep laboratory followed by simultaneous polysomnographic and actigraphic recordings throughout the night. PARTICIPANTS: 21 healthy volunteers. SETTING: A sleep laboratory INTERVENTIONS: None. RESULTS: Ninety-one percent of all PSG epochs were correctly identified by both algorithms, and this accuracy is reasonably satisfactory. The actigraphy was a sensitive method, with values of 99% and 97% for Cole's and Sadeh's algorithms, respectively. However, actigraphy had a low specificity: 34% and 44% for Cole's and Sadeh's algorithms, respectively. The Bland and Altman technique showed that actigraphy systematically overestimated Sleep Latency, Total Sleep Time and Sleep Efficiency while it underestimated Intermittent Awakenings. CONCLUSIONS: The results of this study show the utility of actigraphy as a useful method for assessment of sleep, despite its limitations regarding identification of waking epochs during sleep. The Bland and Altman concordance technique was revealed to be a powerful tool to evaluate how well actigraphy agreed with polysomnography. This technique, combined with calculations of sensitivity and specificity, appears to be the most adequate procedure for the assessment of concordance. PMID- 12627738 TI - Automatic sleep-wake and nap analysis with a new wrist worn online activity monitoring device vivago WristCare. AB - STUDY OBJECTIVE: Vivago WristCare is a new activity monitoring device, which allows long-term online monitoring of the activity of the user. This study evaluates the ability of the device to discriminate sleep/wake patterns during nighttime and during napping. DESIGN: Participants spent one night in the sleep laboratory where signals from polysomnography, actigraphy and WristCare were acquired. In addition, actigraphy and WristCare were used for 3-6 days for nap analysis. SETTING: NA. PATIENTS OR PARTICIPANTS: Participants were 32 adults aged 26-89 years. The participants were studied in three study groups: all subjects, senior subjects (age > 65 years) and middle-aged subjects (age < 65 years). INTERVENTIONS: NA. RESULTS: Sleep/wake patterns were extracted from polysomnography, actigraphy and WristCare for the night slept in sleep laboratory. The agreement percents between the scorings of polysomnography and actigraphy, and between polysomnography and WristCare were about 80 % for all study groups. As total sleep time was estimated and the algorithm was optimized for this measure, the performance of the WristCare and actigraphy were similar. Both actigraphy and WristCare overestimated appreciably total sleep time (TST). Also in nap-analysis, actigraphy and WristCare performed similarly as the number of naps and the length of the naps were compared. CONCLUSIONS: The performance of the WristCare can be assumed to be well comparable to actigraphy in sleep/wake studies. The study suggests that the device may be used in long-term monitoring of sleep/wake patterns with similar performance to actigraphy. PMID- 12627739 TI - Oximeter's acquisition parameter influences the profile of respiratory disturbances. AB - STUDY OBJECTIVES: Pulse oximetry (Sp02) is a key parameter monitored during polysomnographic studies, and different acquisition settings can be employed to obtain this data. The purpose of this study was to determine if the use of different settings would significantly influence scoring of respiratory disturbance events (RDE). DESIGN: Prospective study SETTING: Sleep Disorders Center - community PATIENTS: 30 patients had three identical oximeters simultaneously attached to the digits during polysomnography, each placed in a different recording setting: 3, 6 and 12 seconds. INTERVENTIONS: None. MEASUREMENTS: RDEs were identified by changes in snoring and flow then sub categorized as RDE0, RDE1-2 and/or RDE3 if less than 1%, greater than 1 but less than 3%, and 3% or greater oxyhemoglobin desaturation occurred. Each event was given three labels according to the level of desaturation seen on each oximetry tracing. RESULTS: Significant differences in the mean frequency of RDE types at each recording setting were noted (p < .001). A survey of sleep practitioners revealed changes in clinical behavior when presented examples of such differences. CONCLUSION: These data confirm the impact of different oximetric recording settings on the profile of RDEs and the importance of reporting such acquisition settings in studies of sleep disordered breathing. PMID- 12627740 TI - Skills training versus health education to prevent STDs/HIV in heterosexual women: a randomized controlled trial utilizing biological outcomes. AB - We compared the effectiveness of two different 16-session group interventions for reducing new STD infection among heterosexual women. Two hundred twenty-nine at risk heterosexual women were randomly assigned to skills training (ST) based on the relapse prevention model or health education (HE). Participants were monitored during the year following intervention for STD acquisition, self reports of sexual behavior, and risk reduction skills. Participants in the ST intervention were significantly less likely to be diagnosed with a STD in the year following intervention and demonstrated superior risk reduction skills at 12 month follow-up. Both conditions showed statistically significant reductions in self reports of risky sexual behavior following intervention and at 12-month follow-up. In this sample, the ST intervention was superior to HE for reducing STD acquisition. PMID- 12627741 TI - Effectiveness of a risk reduction intervention among African American women who use crack cocaine. AB - The primary objective of this study was to evaluate the effectiveness of an HIV intervention for African American women who use crack cocaine. Two hundred sixty five women (aged 18-59 years) were randomly assigned to one of two enhanced intervention conditions or to the National Institute on Drug Abuse standard condition. A substantial proportion of women reported no past 30-day crack use at 6-month follow-up (100%-61%, p < .001). Significant (P < .05) decreases in the frequency of crack use; the number of paying partners; the number of times vaginal, oral, or anal sex was had with a paying partner; and sexual risks, such as trading sex for drugs, were reported over time. Significant (p < .05) increases in male condom use with sex partners were observed, as well as decreases in casual partners' refusal of condoms. Findings suggest that combined components of our culturally appropriate, gender-tailored intervention may be most effective at enhancing preventive behavior among similar populations. PMID- 12627742 TI - Evaluation of the PSAMAO "Roulez Protege" mass media campaign in Burkina Faso. Prevention du SIDA sur les Axes Migratoires de l'Afrique de l'Ouest. AB - This study assesses the impact of the Prevention du SIDA sur les Axes Migratoires de l'Afrique de l'Ouest Roulez Protege mass media campaign in Burkina Faso on truckers' likelihood of talking with colleagues about HIV/AIDS, using condoms with an occasional sexual partner, and intending to use a condom in the future. A total of 764 and 1,032 truckers were interviewed in 1997 and 2000 along the Ouagadougou-Niangoloko trucking route. Truckers' exposure to the program has been high and the main message promoted has been greatly understood among those exposed. Logistic regression analyses suggest that the campaign did not have direct behavioral impacts. Exposure to radio spots and participation in group discussions were found to be significantly associated with truckers discussing AIDS with peers or reporting intentions to use a condom in the future. Exposure to Roulez Protege messages through television or billboards was not found to be significantly associated with interpersonal discussion or condom use intentions. Programs should seek to sustain and expand the indirect effects of these types of programs in West Africa. Reinforcing interpersonal counseling interventions should, down the road, lead to overall knowledge and behavior changes among the high-risk trucker population. PMID- 12627743 TI - Interventions to reduce HIV/AIDS stigma: what have we learned? AB - This article reviews 22 studies that test a variety of interventions to decrease AIDS stigma in developed and developing countries. This article assesses published studies that met stringent evaluation criteria in order to draw lessons for future development of interventions to combat stigma. The target group, setting, type of intervention, measures, and scale of these studies varied tremendously. The majority (14) of the studies aimed to increase tolerance of persons living with HIV/AIDS (PLHA) among the general population. The remaining studies tested interventions to increase willingness to treat PLHA among health care providers or improve coping strategies for dealing with AIDS stigma among PLHA or at-risk groups. Results suggest some stigma reduction interventions appear to work, at least on a small scale and in the short term, but many gaps remain especially in relation to scale and duration of impact and in terms of gendered impact of stigma reduction interventions. PMID- 12627744 TI - Reasons for HIV disclosure or nondisclosure to casual sexual partners. AB - The purpose of this study was to investigate reasons HIV-positive gay men give for disclosing or not disclosing their serostatus to their casual sexual partners. Participants were 78 HIV-positive gay men who were part of a larger HIV and disclosure project. A clear factor structure for disclosure emerged which suggests that issues of responsibility dominated men's decisions to disclose. No clear factor structure for nondisclosure emerged. Reasons for disclosure or nondisclosure to casual sexual partners were varied and this data could provide new insights for secondary prevention efforts. More research needs to be conducted to better understand salient issues in considering whether to disclose. PMID- 12627745 TI - Depression and HIV risk behavior among Seattle-area injection drug users and young men who have sex with men. AB - Psychological depression has been identified as a condition that may influence HIV risk behavior among injection drug users (IDUs) and men who have sex with men (MSM). In two Seattle studies, 1,228 IDUs and 429 young MSM completed the Center for Epidemiologic Studies Depression Scale (CES-D); the relationship between depression and injection and sexual risk behavior was assessed using logistic regression analysis. Forty-seven percent of IDUs had CES-D scores > or = 23; a high score was significantly related to injection with a syringe used by another IDU (adjusted odds ratio 1.4) but not other injection risk behavior. Among MSM, CES-D scores > or = 16 were related to reporting 3 or more sex partners in the last 6 months but not to other sexual risk behavior. This analysis suggests that psychological depression may influence certain HIV risk behavior in young MSM and IDUs, and that interventions addressing depression may be indicated. PMID- 12627746 TI - Responses of male inmates to primary partner requests for condom use: effects of message content and domestic violence history. AB - Many women at high risk for HIV infection face resistance and, in some cases, violence as a response to their requests for condom use. The current study investigated how domestically violent and nonviolent men reacted to various condom negotiation approaches. Ten different scenarios, in which the partner provides a justification for a condom request or the context suggests one, were presented to 84 male inmates selected at random from a county jail. As predicted, condom scenarios factored into groupings with content suggestive of high and low relationship threat. Of the justifications presented, yeast infections generated more favorable responses than standard HIV prevention messages. The riskiest condom negotiation scenario was one that suggested infidelity on the part of the woman. Level of male violence severity in the relationship predicted more coercive responses to suggestions of a woman's infidelity. The results suggest that creative strategies that do not call into question the fidelity or commitment of either partner may be more effective in getting men to use condoms and/or to not react violently. PMID- 12627747 TI - Lipoprotein(a): an emerging cardiovascular risk factor. AB - Lipoprotein(a) is a cholesterol-enriched lipoprotein, consisting of a covalent linkage joining the unique and highly polymorphic apolipoprotein(a) to apolipoprotein B100, the main protein moiety of low-density lipoproteins. Although the concentration of lipoprotein(a) in humans is mostly genetically determined, acquired disorders might influence synthesis and catabolism of the particle. Raised concentration of lipoprotein(a) has been acknowledged as a leading inherited risk factor for both premature and advanced atherosclerosis at different vascular sites. The strong structural homologies with plasminogen and low-density lipoproteins suggest that lipoprotein(a) might represent the ideal bridge between the fields of atherosclerosis and thrombosis in the pathogenesis of vascular occlusive disorders. Unfortunately, the exact mechanisms by which lipoprotein(a) promotes, accelerates, and complicates atherosclerosis are only partially understood. In some clinical settings, such as in patients at exceptionally low risk for cardiovascular disease, the potential regenerative and antineoplastic properties of lipoprotein(a) might paradoxically counterbalance its athero-thrombogenicity, as attested by the compatibility between raised plasma lipoprotein(a) levels and longevity. PMID- 12627748 TI - The renal kallikrein-kinin system: its role as a safety valve for excess sodium intake, and its attenuation as a possible etiologic factor in salt-sensitive hypertension. AB - The distal tubules of the kidney express the full set of the components of the kallikrein-kinin system, which works independently from the plasma kallikrein kinin system. Studies on the role of the renal kallikrein-kinin system, using congenitally kininogen-deficient Brown-Norway Katholiek rats and also bradykinin B2 receptor knockout mice, revealed that this system starts to function and to induce natriuresis and diuresis when sodium accumulates in the body as a result of excess sodium intake or aldosterone release, for example, by angiotensin II. Thus, it can be hypothesized that the system works as a safety valve for sodium accumulation. The large numbers of studies on hypertensive animal models and on essential hypertensive patients, particularly those with salt sensitivity, indicate a tendency toward the reduced excretion of urinary kallikrein, although this reduction is modified by potassium intake and impaired renal function. We hypothesize that the reduced excretion of the renal kallikrein may be attributable to a genetic defect of factor(s) in renal kallikrein secretion process and may cause salt-sensitive hypertension after salt intake. PMID- 12627749 TI - What can we infer from double dissociations? PMID- 12627750 TI - Covert recognition and the neural system for face processing. AB - In this viewpoint, we discuss the new evidence on covert face recognition in prosopagnosia presented by Bobes et al. (2003, this issue) and by Sperber and Spinnler (2003, this issue). Contrary to earlier hypotheses, both papers agree that covert and overt face recognition are based on the same mechanism. In line with this suggestion, an analysis of reported cases with prosopagnosia indicates that a degree of successful encoding of facial representations is a prerequisite for covert recognition to occur. While we agree with this general conclusion as far as Bobes et al.'s and Sperber and Spinnler's data are concerned, we also discuss evidence for a dissociation between different measures of covert recognition. Specifically, studies in patients with Capgras delusion and patients with prosopagnosia suggest that skin conductance and behavioural indexes of covert face recognition are mediated by partially different mechanisms. We also discuss implications of the new data for models of normal face recognition that have been successful in simulating covert recognition phenomena (e.g., Young and Burton, 1999, and O'Reilly et al., 1999). Finally, in reviewing recent neurophysiological and brain imaging evidence concerning the neural system for face processing, we argue that the relationship between ERP components (specifically, N170, N250r, and N400) and different cognitive processes in face recognition is beginning to emerge. PMID- 12627751 TI - Perspectives on prosopagnosia and models of face recognition. AB - The two papers by Bobes et al. (2003, this issue) and by Sperber and Spinnler (2003, this issue) add to the large body of literature demonstrating covert face recognition in prosopagnosia. This viewpoint will offer some perspectives on this interesting phenomenon. First, a re-analysis of the empirical literature will indicate an important misconception concerning the preserved abilities of prosopagnosics. The second section will briefly assess the contribution of Bobes et al. (2003, this issue) and Sperber and Spinnler (2003, this issue) to the debate about the locus, in cognitive terms, of the underlying causal deficit in prosopagnosia with covert face recognition. Both papers make reference to the two main models seeking to explain this phenomenon: the model proposed by Burton and colleagues (Burton et al., 1991; Burton and Young, 1999; Young and Burton, 1999) and that proposed by Farah and colleagues (Farah et al., 1993; O'Reilly and Farah, 1999). Finally, an observation will be offered concerning representations of faces in the Burton et al. (1991) model. PMID- 12627752 TI - Covert matching of unfamiliar faces in a case of prosopagnosia: an ERP study. AB - In addition to their deficit in overt face recognition, patients with prosopagnosia also have difficulties in matching sequentially presented unfamiliar faces. Here we assessed the possibility that covert matching of faces was present in a case with prosopagnosia using event-related potentials (ERPs). The participants (patient FE and normal controls) were challenged with a face identity matching task, in which they decided whether two sequentially presented photographs of unfamiliar faces represented the same person. Only internal face features were used and the two faces in a pair differed in emotional expression. FE failed to overtly match these stimuli. In contrast, the ERPs revealed evidence of covert matching. If the two faces within a pair of stimuli depicted different posers, then the response to the second face contained an enhanced N300 compared to the situation where the identity of the faces was the same. The latency of the N300 was the same as a similar component found in controls. These results suggest that some cases with prosopagnosia have a covert ability to match unfamiliar faces, with similar temporal dynamics as controls, which in contrast with the idea that a generalized slowing of face processing occurs in all cases of prosopagnosia. PMID- 12627754 TI - Interhemispheric integration of letter stimuli presented foveally or extra foveally. AB - Two experiments are reported in which participants decided whether a single target letter presented below and to the left or right of fixation matched either of two probe letters presented above the fixation cross to the left and right. The level of matching required was either physical (A-A) or abstract (A-a). All three letters were presented either extra-foveally (Experiment 1) or within the fovea (Experiment 2). In both experiments, physical matching was faster than abstract matching. Physical matching was faster within than across visual fields while abstract matching showed the opposite pattern. Matching was faster when the matching probe letter was in the LVF than when it was in the RVF. Importantly, the pattern of results was the same for extra-foveal and foveal presentations, supporting the theory that the representation of the fovea is split down the middle into two visual fields rather then being bilateral. The practical implication following this study is that lateralization studies can be performed with closer to fixation stimuli presentation. PMID- 12627753 TI - Covert person recognition: its fadeout in a case of temporal lobe degeneration. AB - Covert person recognition was investigated longitudinally over a three-year period in a patient suffering from "Crossmodal Familiar Person Agnosia", possibly due to a fronto-temporal dementia in its right temporal variant (Gentileschi et al., 2001). The progressive neuronal degeneration in the cortical regions critical for face recognition (viz., right infero-temporal areas) presented us with the opportunity to check Burton et al.'s (1991) and Farah et al.'s (1993) hypothesis on the dissociation between overt and covert face recognition in a neuropsychological condition which, however, is neurologically and cognitively different from that of focal "associative prosopagnosia". Covert person recognition starting from overtly unrecognised faces was assessed by means of learning tasks of face/name association involving celebrities. It was assumed that some unconsciously spared information would selectively enhance the relearning rates when famous faces were paired with their true names. In fact, the true-name advantage (i.e., selective saving for experimental relearning of true name pairings) reached significance at first assessment, carried out five years from clinical onset. Effect faded away two and three years later on, thus abolishing the overt/covert dissociation in face recognition. These findings support Burton et al.'s (1991) and Farah et al.'s (1993) hypothesis of covert face recognition as the consequence of partial and incomplete activation of person semantics, due, in the present case, to the impoverishment of Gentileschi et al.'s (2001) "exemplar semantics" storehouse. Moreover, it turned out that covert recognition does not imply a different learning slope, but an overall different level of the learning profile. PMID- 12627755 TI - The residual calculation abilities of a patient with severe aphasia: evidence for a selective deficit of subtraction procedures. AB - We report the case study of a severe fluent aphasic patient, who showed relatively preserved numerical abilities. A detailed investigation of number processing indicated good numerical comprehension and a relative sparing of addition and subtraction abilities; on the other hand, multiplication and division were severely impaired. A further study of multi-digit operations showed that the patient's performance was characterized by a selective impairment of the borrowing procedure, in which she applied the so-called Smaller-from-Larger bug, typically observed in children learning to calculate. The present case provides further evidence for the dissociation between operations based on verbal sequences and on quantity manipulation, respectively impaired and preserved in patients with severe aphasia. Moreover, it provides evidence indicating that procedures may be dissociated from conceptual knowledge within a single arithmetical operation. PMID- 12627756 TI - Verbal free recall in high altitude: proper names vs common names. AB - Exposure to extreme altitude is known to cause a general impairment of cognitive functions. In this study we investigated the effect of high altitude on the recall of supraspan lists of proper and common names. High altitude seems to have a dramatic effect on the recall of proper names, while common names are more resistant to hypoxia. This effect, however, seems to selectively concern the early items of the lists. This damage to the primacy effect in proper name recall seems permanent over time. The absence of damage in recalling names from the last serial position clearly shows how STM functions are relatively more resistant to the lack of cognitive sources caused by hypoxia. PMID- 12627757 TI - A ticklish question: does magnetic stimulation of the primary motor cortex give rise to an 'efference copy'? AB - In order to make accurate predictions about the somatosensory consequences of our own movements, an 'efference copy' of motor output appears to be used (probably in the cerebellum) as a basis for anticipating and canceling incoming information about self-produced movement. One everyday phenomenon that results from these predictive processes is that of being unable to tickle oneself. We examined whether involuntary movements induced by magnetic stimulation of (a) the motor cortex and (b) the bicep muscle, both resulting in standard stimulation of the participant's bare foot, would be perceived as ticklish. In both cases, the stimulus to the foot was rated as ticklish as when the experimenter tickled the participant's foot (using an identical stimulus). We conclude that magnetic stimulation of the motor cortex is unlikely to give rise to an efference copy of motor output. PMID- 12627758 TI - Cortical representation of the fovea: implications for visual half-field research. PMID- 12627759 TI - Should the temporal cortex be chopped in two? PMID- 12627760 TI - Double dissociations: not magic, but still useful. PMID- 12627761 TI - The emergence of a double dissociation in the modulation of a single control parameter in a nonlinear dynamical system. PMID- 12627762 TI - One word, one module? PMID- 12627763 TI - Interpreting double dissociations in connectionist networks. PMID- 12627764 TI - Dissociation in connectionist systems. PMID- 12627765 TI - Lesion studies, spared performance, and cognitive systems. PMID- 12627766 TI - Double dissociations, models and serial position curves'. PMID- 12627767 TI - Pitfalls in the method of double dissociation: delineating the cognitive functions of the hippocampus. PMID- 12627768 TI - Double dissociation and the benefit of experience. PMID- 12627770 TI - The module mistake. PMID- 12627769 TI - Double dissociations in developmental disorders? Theoretically misconceived, empirically dubious. PMID- 12627771 TI - How much can we learn from double dissociations? PMID- 12627772 TI - More on the not-the-liver fallacy: medical, neuropsychological, and perceptual dissociations. PMID- 12627773 TI - Interpreting dissociations: the issue of task comparability. PMID- 12627774 TI - The elusive dissociation. PMID- 12627775 TI - Process decomposition from double dissociation of subprocesses. PMID- 12627776 TI - Beyond existence: inferences about mental processes from reversed associations. PMID- 12627777 TI - Inference and explanation in cognitive neuropsychology. PMID- 12627778 TI - Dissociations: double or quits? PMID- 12627779 TI - Beyond task dissociation logic: a richer conception of cognitive neuropsychology. PMID- 12627780 TI - Acute appendicitis. PMID- 12627781 TI - Case of the month. Islet cell tumor of the pancreas. PMID- 12627782 TI - Cardiac multidetector-row computed tomography: initial experience using 16 detector-row systems. PMID- 12627783 TI - Imaging neuroblastoma in children. AB - Neuroblastoma is a common solid tumor of childhood that can involve the abdomen, thorax, pelvis, or the head and neck. The clinical manifestations are dependent on the widespread distribution of neural crest tissue and the length of the sympathetic chain involvement. Abdominal pain and hypertension may occur as a result of renal vasculature compression; respiratory distress may be evident in thoracic tumors; and Homer's syndrome or heterochromia of the iris may manifest from neuroblastoma of the head and neck. In addition, symptoms of cord compression and back pain may result from spinal cord compromise due to epidural invasion. Metastatic involvement of the liver, skin, periorbital regions, or bone may cause hepatomegaly, skin nodules, proptosis, or bone marrow failure, respectively. Clinical findings along with tumor metastasis may be studied by various imaging modalities to assess the nature and extent of the tumor. Diagnostic tests include plain radiography, ultrasonography, CT scanning, and MR imaging. Bone marrow studies, bone scans, and scintigraphy with 131I metaiodobenzylmandelic may be utilized for metastatic evaluation. By using these imaging studies to detect the nature and behavior of neuroblastoma, early intervention may indeed improve patient survival. PMID- 12627784 TI - An innovative surgical suture and needle evaluation and selection program. AB - This report describes an innovative suture and needle clinical evaluation program jointly designed by hospital representatives of Consorta, Inc., a healthcare resource management and group purchasing organization, and United States Surgical/Davis & Geck Sutures (USS/D&G), manufacturer of surgical biomaterials. Nineteen Consorta shareholder hospitals enrolled 699 surgeons to participate in Phase I of this nonexperimental observational study of the clinical performance of surgical needles and sutures. Performance characteristics of the sutures and needles produced by USS/D&G, which were evaluated in 3407 surgical procedures, included packaging and ease of opening, needle strength and sharpness, tissue drag, knot security, tensile strength, and clinically acceptable and unacceptable determinations. In these 30-day studies, the surgeons concluded that the needles and sutures were clinically acceptable in 98.1% of the evaluations. The general, cardiothoracic, and orthopedic surgeons, who performed 73.8% of the product evaluations, reported that the suture and needle products were clinically acceptable in 97.2% of the evaluations. More than half (50.1%) of the evaluations involved the POLYSORB* braided synthetic sutures,which received a clinically acceptable rating in 98.4% of the evaluation. The next most frequently used sutures were the SOFSILK*, followed by the monofilament nylon suture. SOFSILK* was found to be clinically acceptable in 98.7% of the evaluations, whereas the monofilament nylon was noted to be clinically acceptable in 96.3% of the evaluations. Surgical needles made by USS/D&G had a 97.9% clinical acceptability rating. PMID- 12627785 TI - Phosphorylcholine-coated stents. AB - Stainless steel stents have been developed to perform their primary purpose of providing sufficient physical support to an artery following percutaneous transluminal coronary angioplasty (PTCA) in order to prevent vessel recoil. Early clinical problems, largely caused by subacute thrombosis (SAT), have been significantly reduced with a new generation of antiplatelet agents. Despite improved pharmacological control of SAT in many patients, the potential for stent occlusion remains a life-threatening problem for those with more complex lesions. One strategy for dealing with this is to improve the hemocompatibility of the stent surface with coatings. A phosphorylcholine (PC)-based stent coating has been designed for this purpose; we describe in detail the evaluation of the device, in vitro, through preclinical and early clinical data. We discuss studies that demonstrate that the PC-coated stent may offer some clinical advantage over uncoated counterparts. The extensive experience with this PC-coated device supports its nonthrombogenic nature, excellent acceptance by arterial tissue, and long-term stability in vivo. These characteristics, together with its ability to load and release a variety of therapeutic agents, also make the device a natural choice of platform for future stent-mediated drug delivery for treating restenosis. PMID- 12627786 TI - The need for population-based epidemiology studies in the United States. PMID- 12627787 TI - A nationwide followup study of autoimmune and connective tissue disease among hip and knee implant patients. AB - Because implants can provoke varied immune system responses, we assessed whether hip and knee implant recipients had an increased risk of autoimmune/connective tissue diseases (AI/CTDs). Using national registry data from Sweden, we compared hospitalization rates for AI/CTD in 101,771 hip and 23,891 knee implant recipients to rates in the general population. Hip patients were followed up to 22 years and knee patients up to 14 years postimplantation. Our findings indicate that it is unlikely that hip or knee implantation results in any increased risk for most AI/CTDs. After long-term followup, the associations we observed with polyarteritis nodosa and fibrositis could be the basis for future investigations. PMID- 12627788 TI - Cancer risk among pacemaker recipients in Denmark, 1982-1996. AB - To address concerns about the potential carcinogenicity of pacemakers, we launched the first epidemiologic study of cancer incidence among pacemaker recipients. A nationwide cohort of 16,357 pacemaker recipients in Denmark from 1982 through 1996 was identified. The Danish Cancer Registry was used to identify all incident cancers within the cohort, with almost 75,000 person-years of observation. The cohort had a slight excess of cancer overall (SIR = 1.19, 95% confidence interval [95% CI, 1.1-1.2]). This was largely caused by an elevated SIR for multiple myeloma among men (SIR = 1.78,95% CI, 1.1-2.8), which increased to 2.60 (95% CI, 0.9-5.7) 5-9 years after implantation, and to a similarly increased SIR for kidney cancer among women (SIR = 2.05, 95% CI, 1.3-3.0), which increased to 3.39 (95% CI, 1.6-6.2) after a latency period of 5-9 years. An excess of urinary bladder cancer was also seen after 10 years. No excess risk was observed for breast cancer or sarcomas, although the SIRs for sarcomas tended to increase over time, based on small numbers. Our results are largely reassuring but, as pacemakers become more common and are implanted at earlier ages and as survival following implantation improves, the excesses of bladder cancer, multiple myeloma among men, and kidney cancer among women with long-term followup warrant further investigation. PMID- 12627789 TI - Incidence of breast and other cancers among Finnish women with cosmetic breast implants, 1970-1999. AB - Epidemiologic evidence does not point to a carcinogenic effect of silicone implants on the breast, and evidence for or against a carcinogenic effect at sites other than the breast is limited. To examine subsequent cancer risk among women with cosmetic breast implants, we conducted a cohort study of 2171 women in Finland identified from operation diaries of major hospitals and private clinics, 1970-1999. The nationwide population and health outcome registries in Finland were used to trace these women for cancer incidence through 1999. Standard statistical techniques were used to compute expected values based on general population rates. The measure of risk was taken as the ratio of observed to expected cancers, that is, the standardized incidence ratio (SIR) and its 95% confidence interval (CI). Among the 2171 women with cosmetic breast implants, 30 developed cancer against 33.7 expected (SIR = 0.9,95% CI = 0.6-1.3). There was no evidence for an increase in breast cancer risk (SIR 0.5, 95% CI, 0.2-1.0), even among those followed for more than 10 years (2 observed, 4.6 expected). Stage at breast cancer diagnosis did not differ from that expected nor did incidence of any other cancer. Although hindered by small numbers, the consistency of our results with those of other Nordic studies leads us to conclude that cosmetic breast implants are not a cause of cancer and that they do not appear to delay the detection of breast cancers. PMID- 12627790 TI - An in vitro study of the effect of in-folds on the durability of mammary implants. AB - We hypothesize that the presence of in-folding in a filled mammary shell leads to shortening of the time to failure under tensile fatigue. Specimens, cut from pristine shells, were mounted in S-folded, creased, and unfolded configurations. A characteristic change, occurring in a transmembrane capacitance-proportional alternating current (AC) signal, which was used to monitor working of each specimen, was chosen as a robust marker that preceded frank shell perforation. Subjecting all specimens to stringent, controlled conditions led to time-to fatigue failure estimates that demonstrated an order-of-magnitude reduction in lifetime for the folded specimens when compared with that for pair-matched, unfolded specimens. Creases also reduced the fatigue lifetime, but not to the degree that folds did. Observations based on the experimental behavior of folded shell material provide possible mechanisms for the potential lifetime clinical reduction of in-folded implants as well as for the development of silicone-on silicone abrasive wear and generation of debris. PMID- 12627791 TI - Allergic reaction to platinum in silicone breast implants. AB - Platinum is used as a catalyst in the manufacture of silicone breast implants. Because platinum is recognized as a potent sensitizer in certain circumstances, some have expressed concern that women with silicone breast implants are exposed to platinum, which is causing allergic reactions. We searched the literature for information on the level of platinum in breast implants and reports of sensitization that clearly related to platinum in women with breast implants. We found no published report with convincing evidence that platinum causes allergic reactions in women with breast implants or that women with breast implants are any more likely to have allergic reactions than women without breast implants. PMID- 12627792 TI - Craniofacial skeletal deviations following in utero exposure to the anticonvulsant phenytoin: monotherapy and polytherapy. AB - OBJECTIVE: To identify and quantify the craniofacial effects from prenatal exposure to phenytoin monotherapy and polytherapy using cephalometric, hand wrist, and panoramic radiographs and to determine if such deviations persist with age. DESIGN: Craniofacial structures of 28 anticonvulsant-exposed individuals were evaluated using 20 landmarks in lateral cephalometric radiographs and 19 landmarks in frontal cephalometric radiographs. Skeletal maturity was assessed using hand-wrist radiographs. Dental maturity and the presence of dental anomalies were evaluated using panoramic radiographs. Eleven individuals were re evaluated 7 years later, on average, to determine the persistence of any measured deviations. SETTING AND SAMPLE POPULATION: Department of Growth and Development, Harvard School of Dental Medicine and Massachusetts General Hospital. Patients were recruited from several sources. OUTCOME MEASURE: The evaluated dimensions included linear, angular, and proportional measures. RESULTS: The most common deviations were decreased height and length of the maxilla, decreased length of the posterior cranial base, length of the mandible, cranial width and level of the cribriform plate, and a decrease in the Wits Appraisal assessment. The deviations were more significant in the polytherapy-exposed individuals than in the monotherapyexposed individuals. These deviations, especially in the maxilla, persisted with age as revealed in a re-evaluation of 11 individuals. CONCLUSION: The craniofacial skeletal findings among individuals exposed in utero to phenytoin monotherapy or phenytoin polytherapy, when considered in aggregate, suggest a mild pattern of maxillary hypoplasia that becomes more pronounced with age. PMID- 12627793 TI - A stochastic evaluation of the decision to specialize in orthodontics. AB - OBJECTIVE: The objective of this paper is to identify under different scenarios, and from a financial point of view, the conditions required to successfully switch from the general dentistry practice to orthodontics. STUDY DESIGN: A mail survey was used to collect the data from the practicing orthodontists. They estimated the income, at certain points, in the working life of an orthodontist. The general practitioner data were taken from the American Dental Association figures. Subsequently, a stochastic model was constructed. RESULTS AND CONCLUSION: Those who decide to buy an existing practice expect higher profits in the near future, and therefore the required minimum number of remaining years of practice is lower than for those deciding to start a new practice. For both scenarios, the 3-year residency will delay the profits compared with a 2-year residency. Thus, an increased number of remaining years of practice is required. There must be more than 10 working years left in the practitioner's life to make the switch profitable. PMID- 12627795 TI - Craniofacial anthropometry of Hong Kong Chinese babies: the eye. AB - OBJECTIVES: To provide a database of the canthal measurements of Chinese infants born in Hong Kong. DESIGN: Prospective cross-sectional study. SETTING AND SAMPLE POPULATION: A total of 2,371 healthy singleton, born consecutively at the Prince of Wales Hospital and the Union Hospital from June 1998 to June 2000, were included in the study. The range of gestation was 33-42 weeks. MEASUREMENTS AND RESULTS: Canthal parameters were measured and included the outer canthal distance (OCD), intercanthal distance (ICD) and palpebral fissure length (PFL). The data generally show higher values for males in the parameters measured. The measurements were also compared with data published overseas. PFL and OCD were significantly larger in Hong Kong Chinese newborn babies, whereas ICD was smaller. CONCLUSION: This study establishes the first set of gestational age specific standard of such craniofacial parameters for Chinese newborn babies, potentially enabling early syndromal diagnosis. There are significant inter racial differences in these canthal parameters. PMID- 12627794 TI - Expression of the retinoblastoma family of tumor suppressors during murine embryonic orofacial development. AB - OBJECTIVE: To elucidate the role of the retinoblastoma (Rb) family of tumor suppressors and growth regulators in transforming growth factor beta (TGFbeta) mediated embryonic palatal growth and morphogenesis. DESIGN: The spatio-temporal expression patterns of the RB1, RB2/p130, and p107 tumor suppressor genes, their gene products (pRb, p130 and p107) and phosphoforms were examined in the developing murine secondary palate utilizing reverse transcriptase polymerase chain reaction (RT-PCR) and immunoblot/immunolocalization analyses with phospho specific antibodies. RESULTS: The RB1, RB2/p130, and p107 tumor suppressor genes and their gene products (pRb, p130, and p107) were differentially expressed in embryonic palatal tissue during the critical period of secondary palate development [gestational days (GD) 12-14]. Both hyper- (115 kDa) and hypo phosphorylated (110 kDa) forms of pRb were expressed, with a notable transient decrease in expression on GD 13. Functional (hypo-phosphorylated) forms of pRB predominated during the critical period of palatogenesis. As opposed to pRb expression, p130 expression was transiently elevated on GD 13 in the embryonic palate, and functional (hypo-phosphorylated) forms were expressed at exceedingly low levels. p107 levels gradually declined over the course of palatogenesis. This diminution in expression, however, was accompanied by a transition to more functional (hypo-phosphorylated) forms of the p107 protein. When compared with Rb expression patterns in the whole embryo/fetus, each of the three Rb proteins exhibited unique and specific temporal patterns of expression in the developing palate. RT-PCR analyses of RB1, RB2/p130, and p107 mRNA expression in embryonic palatal tissue revealed patterns of expression which paralleled steady-state protein levels of pRb, p130, and p107 in the developing tissue. Immunolocalization of the Rb proteins demonstrated ubiquitous expression of pRb, p130, and p107 in embryonic palate mesenchyme and epithelium during GD 12-14 of development with intense nuclear staining of the Rbs in palate epithelial cells on days 12 and 13 of gestation. CONCLUSION: Changing patterns of pRb, p130, and p107 protein expression and phosphorylation were evident in the embryonic secondary palate during the course of palatal ontogenesis. As the function of all three Rb proteins is regulated primarily through their phosphorylation, this suggests significant variation in Rb protein functionality during the course of palate development. Additional evidence from our laboratory (manuscript in preparation) indicates that the TGFbetas, key regulators of palatal growth and morphogenesis, are able to differentially modulate phosphorylation of the Rb proteins in this developing tissue, and hence the TGFbetas may regulate the function(s) of the Rb proteins during palatal ontogenesis. Although functionality of the Rb proteins is known to be regulated primarily through post-translational modification (i.e. phopsphorylation), comparison of RB1, RB2/p130, and p107 steady-state mRNA levels with pRb, p130, and p107 steady-state protein levels in the developing palate levels suggests a significant degree of regulation at the transcriptional level. Differential patterns of Rb expression and phosphorylation in the developing palate suggest that pRb, p107, and p130 may each play unique roles in various aspects of growth, morphogenesis and cellular differentiation during palatal ontogenesis. Studies directed at elucidating the precise cellular role(s) of pRb, p107, and p130 during palate development, and at TGFbeta regulation of Rb expression/phosphorylation are ongoing in our laboratory. PMID- 12627796 TI - Craniofacial morphology and dental age in children with Silver-Russell syndrome. AB - OBJECTIVES: This investigation is a part of a multidisciplinary descriptive evaluation of the Silver-Russell syndrome (SRS). The aim of this study was to describe the craniofacial morphology, occlusion and dental age in children with the SRS. DESIGN: A descriptive literature-controlled study. SETTING AND SAMPLE POPULATION: Sixteen children diagnosed as having SRS, 10 boys and six girls, aged 4.4-14.5 years, were referred from different parts of Sweden to the Queen Silvia Children's Hospital, Goteborg University. EXPERIMENTAL VARIABLE: Facial morphology was measured on lateral and postero-anterior radiographs. Occlusion, tooth eruption and palatal height were measured on casts, and dental maturity was evaluated on orthopantomograms. OUTCOME MEASURE: Linear and angular measurements were obtained from lateral radiographs and the ratios of the linear measurements from the postero-anterior radiographs. The degree of tooth calcification shown on orthopantomograms was taken as a measure of dental maturity. Biometric measurements were taken and the degree of tooth eruption was recorded from the dental casts. The SRS children were compared with reference groups with t-test and z-scores. RESULTS: Overall, SRS children were found to have smaller linear facial dimensions and deviations in the facial proportions, such as a small retropositioned, and steeply inclined maxilla and mandible, and a proportionally larger anterior facial height in relation to the posterior facial height. In 40% of them a smaller facial height or length on one side (facial asymmetry) was correlated to the smaller side of the body. The frequency of malocclusions was higher, and the palatal height showed a tendency towards an increase. Dental maturity was within normal limits, while the time of tooth eruption was slightly delayed. CONCLUSIONS: The deviating facial morphology described above is a part of the syndrome, which is characterized by short stature, growth hormone deficiency and asymmetries of the body. The higher percentage of malocclusions in the SRS children might lead to a greater need of orthodontic treatment. PMID- 12627797 TI - Association between ratings of facial attractivess and patients' motivation for orthognathic surgery. AB - OBJECTIVES: To compare the judgments of facial esthetics, defects and treatment needs between laypersons and professionals (orthodontists and oral surgeons) as predictors of patient's motivation for orthognathic surgery. DESIGN: Two panels of expert and naive raters were asked to evaluate photographs of orthognathic surgery patients for facial esthetics, defects and treatment needs. Results were correlated with patients' motivation for surgery. SETTING AND SAMPLE POPULATION: Fifty-seven patients (37 females and 20 males) with a mean age of 26.0 +/- 6.7 years were interviewed prior to orthognathic surgery treatment. Three color photographs of each patient were evaluated by a panel of 14 experts and panel of 18 laypersons. EXPERIMENTAL VARIABLES: Each panel of raters were asked to evaluate the facial morphology, facial attractiveness and recommend surgical treatment (independent variables). The dependent variable was the patient's motivation for orthognathic surgery. Outcome measure--Reliability of raters were analyzed using an unweighted Kappa coefficient and a Cronbach alpha coefficient. Correlations and regression analyses were used to quantify the relationship between variables. RESULTS: Expert raters provided reliable ratings of certain morphological features such as excessive gingival display and classification of mandibular facial form and position. Based on the facial photographs both expert and naive raters agreed on facial attractiveness of patients. The best predictors of patients' motivation for surgery were the naive profile attractiveness rating and the patients' expected change in self-consciousness. CONCLUSIONS: Expert raters provide more reliable ratings on certain morphologic features. However, the layperson's profile attractiveness rating and the patients' expected change in self-consciousness were the best predictors for patients' motivation for surgery. These data suggest that patients' motives for treatment are not necessarily related to objectively determined need. Patients' decision to seek treatment was more correlated to laypersons' rating of attractiveness because they see what other laypersons see, and are directly or indirectly affected by others reactions to their appearance. These findings may provide useful information for clinicians in counseling patients who seek orthognathic surgery. PMID- 12627798 TI - Reflections on the question: what does molecular mycology have to do with the clinician treating the patient? AB - After a talk on regulation of gene transcription in Candida albicans, a clinical mycologist was heard to ask: "What difference does all that make to the lady dying of disseminated Candida infection in her hospital bed?" The rapid expansion of research in fungal diseases is widening the communication gap between individuals with responsibility for patient care and those who study pathogenic fungi at the level of molecular biology. DNA-based technologies have produced real advances for patient care by delivering superior methods for fungus identification and strain typing that will soon find a routine place in patient management. Molecular research into the fine detail of the host-pathogen interplay in fungal disease has also made great advances, though the spin-offs to benefit the clinician are not yet obvious. Detection of fungal DNA as a non culture diagnostic method still requires considerable refinement before it can be regarded as a routinely useful approach, and genomic-based strategies for discovery of novel antifungal drugs from molecular targets have so far produced no agents that have entered development. It is inevitable that, in time, several aspects of molecular mycology research will become important basic knowledge for the clinician who is treating the patient. Therefore, clinicians and bench scientists with specialist interest in mycoses need to retain a reasonable level of mutual comprehension and respect of each other's work rather than assuming their professional paths are divergent. PMID- 12627799 TI - Analyses of phagocytosis, evoked oxidative burst, and killing of black yeasts by human neutrophils: a tool for estimating their pathogenicity? AB - The pathogenicity of several dematiaceous yeasts that have, to date, rarely been isolated in humans remains unclear. Because professional phagocytes are prominent in lesions caused by dematiaceous fungi, we address this issue by comparing phagocytosis, evoked oxidative burst and killing by human neutrophils of different black yeasts in vitro. Whereas phagocytosis of all black yeasts tested and evoked oxidative burst yielded comparable results, in contrast, the degree of killing differed significantly after 5 h. Thereby, two groups could be identified; one in which strains are killed at high rates, for example, Hortaea werneckii (81 +/- 11.6%), Exophiala castellanii (96 +/- 8.6%), Phaeoannellomyces elegans (93 +/- 9.7%), Phaeococcomyces exophialae (87 +/- 8.7%), and the other in which strains are killed to a lesser degree, for example, Exophiala dermatitidis (ATCC 34100) (61 +/- 9.5%), E. dermatitidis (CBS 207.35) (66 +/- 7.5%), E. jeanselmei (50 +/- 10.5%), E. mesophila (63 +/- 11.6%), E. bergeri (63 +/- 9.1%), and E. spinifera (57 +/- 9.6%). Non-pigmented yeasts were killed at levels comparable with those at which the white mutant strain of E. dermatitidis (ATCC 44504) was killed (95 +/- 7.5%); the yeast strains tested were Candida albicans (DSM 11943) (95 +/- 4.0% killing) and Saccharomyces cerevisiae (DSM 1333) (95 +/- 10.3%). Comparison of killing rates with the observed pathogenicity of the melanized species suggests that low killing rates might indicate or even predict a high degree of invasiveness. Although previous experiments revealed that melanization conferred killing resistance on E. dermatitidis, the differences in killing rates of other dematious fungi suggest that melanization of the cell wall is in itself insufficient to confer virulence. PMID- 12627800 TI - Pharmacokinetics of amphotericin B in serum and tissues in mice treated with amphotericin B-Intralipid. AB - In our previous research we studied the efficacy of amphotericin B (AMB) Intralipid (AMB-IL) admixtures in the treatment of experimental systemic candidosis in naive and compromised mice. In this study we evaluated the levels of AMB in blood and several organs (kidneys, liver, spleen, lungs and heart) at different times after injection (from 5 min to 14 days). Comparisons were made with levels occurring in animals treated with Fungizone or AmBisome. A sensitive high-pressure liquid chromatography (HPLC) method was adapted for assaying AMB in blood and tissues. We used reversed-phased column and a simple mobile phase consisting of acetonitrile and EDTA. We found that blood levels of AMB in mice injected with AMB-IL were higher than those obtained in animals treated with Fungizone, but similar to those obtained in mice administered AmBisome. Compared with conventional Fungizone therapy, administration of lipid formulations of AMB (both AMB-IL and AmBisome) resulted in higher concentrations of AMB in the liver and spleen, but lower concentrations in the kidneys and lungs. PMID- 12627802 TI - Relationships of Scopulariopsis based on LSU rDNA sequences. AB - Relationships between Scopulariopsis species and allied fungi were studied by sequencing a 350 bp gene region of the large subunit ribosomal RNA gene (LSU rDNA). In addition, a limited morphological dataset of nine characters was included in the cladistic analysis. Nineteen mitosporic strains (nine Scopulariopsis, five Wardomyces, three Doratomyces, one Trichurus and one Scedosporium species) and 21 meiosporic strains (14 Microascus, 4 Kernia and 3 Pithoascus species) were studied. The data were analysed using parsimony methods. Based on the analyses, the studied microascaceous fungi are divided to 12 molecular lineages. Most of the opportunistic human pathogenic Scopulariopsis species are placed in one clade ('Microascus manginii Clade'). Most synnematous anamorphs with Scopulariopsis-like conidia (Doratomyces and Trichurus) are placed in another clade ('Microascus albonigrescens Clade'), together with Wardomyces. Microascus sensu lato can be divided into seven clades which also incorporate all studied Pithoascus, Scopulariopsis, Wardomyces and Trichurus species and most of the Doratomyces species. Most of the Kernia teleomorphs and one Doratomyces species are placed in a different main clade, together with Pseudallescheria and Petriella. Future alternatives in the taxonomy of Microascus include splitting the genus or redefining it to include deviating taxa. More molecular data need to be obtained and considered in either case. PMID- 12627803 TI - Isolation and characterization of an avirulent Candida albicans yeast monomorphic mutant. AB - Mutagenesis of Candida albicans strain ATCC 26555 with N-methyl-nitro-N nitrosoguanidine followed by plating on solid yeast nitrogen base-N acetylglucosamine medium at 37 degrees C yielded colony morphology variants that were characterized as forming smooth colonies, in contrast to the rough colonies formed by the parental strain. One yeast monomorphic mutant, CAL4, was studied in detail. Strain CAL4 is defective in filamentous growth, unable to form hyphae or pseudohyphae in vivo and in vitro. These filamentous structures are not elicited by commonly used external stimuli such as serum. The mutant had no obvious alterations in its mannan, glucan or chitin content. The total quantity of non covalently linked wall proteins was reduced in the mutant strain, but the electrophoretic pattern shown by these proteins was identical to that of proteins from the parental strain. CAL4 showed major differences from the parental strain in its formation of covalently linked wall proteins. An important aspect of these differences lay in the practical absence of proteins recognized by two monoclonal antibodies, 1B12 and 3H8, which are considered valuable tools in the diagnosis of candidiasis in part because they normally react strongly with all strains. The C. albicans mutant, blocked in yeast-mycelium transition, was avirulent in a mouse model, although it was able to grow in animal tissues. PMID- 12627804 TI - Isolation and purification of chiamydospores of Candida albicans. AB - Two methods were investigated for their efficiency in isolating and purifying chlamydospores of Candida albicans. Chlamydospores were disconnected from pseudomycelial cells either enzymatically using beta-glucuronidase or mechanically by ultrasonic treatment. Free chlamydospores were separated from other cell material by sucrose gradient centrifugation. The resulting preparations were inspected by light-microscopy and electron-microscopy. Both methods yielded preparations with a level of over 90% chlamydospore cells. Ultrasonic treatment caused little change to the ultrastructure of the chlamydospores, whereas the enzyme treatment profoundly affected the cell wall. It is concluded that ultrasonic treatment is an efficient method for obtaining pure preparations of chlamydospores. PMID- 12627801 TI - Candida albicans cell wall antigens for serological diagnosis of candidemia. AB - Serological tests for diagnosis of disseminated fungal infections in the immunocompromised host are used with varying results. In the present study, the relative ability of antibodies to specifically recognize Candida albicans cell wall components was evaluated in order to find antigenic markers for serological diagnosis of candidemia. Native C. albicans cell wall fragments (CW), periodate- (CWIO4) and proteinase-K- (CWP) treated CW, a mildly extracted phosphopeptidomannan (PPM), and beta(1-3)(1-6)-glucan were used as antigens in ELISA with sera from rabbits immunized with C. albicans (n = 10), patients with culture proven candidemia (n = 8) and healthy individuals (n = 8). The antibody response in rabbits consisted predominantly of anti-PPM antibodies, a finding that was substantiated by inhibition-ELISA. Consistently, periodate treatment (CW104) destroyed a major proportion of the antigenic epitopes. Low rabbit antibody levels were found against glucan, the major Candida cell wall component. These results supported the conclusion that glucan is localized mainly in the inner part of the C. albicans cell wall. In contrast to rabbits' serum IgG antibody response against PPM, which was at least tenfold higher than that raised against CW, patients with candidemia had similar IgG antibody levels against both antigens. These levels were significantly higher than those seen in healthy controls (CW, P = 0.0005 and PPM, P < 0.0001). Although the human anti-glucan and anti-CWIO4 IgG antibody levels were low overall, they were nonetheless significantly increased in the patient group (P = 0.0159 for antiglucan and P = 0.0491 for anti-CWIO4). In addition, a correlation was noticed between levels of these antibodies. No significant differences were found between patients and controls for IgM antibodies when CW, CWIO4, PPM and Glu were used as antigens. In conclusion, IgG antibodies to PPM and native cell wall fragments (CW) were highly discriminatory for recognition of candidemia and these antigens are thus promising candidates for use in serodiagnosis. PMID- 12627805 TI - Pathogenicity of Cryptococcus neoformans var. gattii in an immunocompetent mouse model. AB - The pathogenicity of two different genomic profiles of Cryptococcus neoformans var. gattii serotype B isolated from goats that died from cryptococcal pneumonia was assessed in an experimental model of immunocompetent mice. One strain of each randomly amplified polymorphic DNA (RAPD) profile (GR52 and GR56) and three reference C. neoformans isolates representing serotypes B, D and C were used. BALB/c male mice were inoculated by the intraperitoneal route with each strain. After 4 weeks of follow-up, the animals were sacrificed and autopsy specimens of testes, liver, spleen, kidney, lungs and brain were cultured and stained for histopathology. Although spontaneous mortality was only 2% (one animal), all mice except for those inoculated with serotype C showed positive cultures in almost one organ. The strain GR52 isolated from goat showed the highest rate of positive cultures (80%) followed by serotype D (77%). Serotype B reference strain and second goat strain GR56 were both isolated from 70% of samples. Serotype C was recovered in only 33% of organs, and never from brain or lung specimens. GR52 grew abundantly from all lung cultures, and yeast cells with large capsules were seen in histopathology inside the alveoli, peribronchial vessels and interalveolar spaces. They appeared to elicit no inflammatory response. We conclude that intraperitoneally inoculated C. neoformans var. gattii shows high virulence in this immunocompetent mouse model. Strain GR52 was highest in pathogenicity and had marked lung tropism. In contrast, the serotype C reference strain showed the lowest pathogenicity and seemed not to spread outside the abdominal viscera. PMID- 12627806 TI - Evaluation of a polymerase chain reaction reverse hybridization line probe assay for the detection and identification of medically important fungi in bronchoalveolar lavage fluids. AB - An assay system in which polymerase chain reaction (PCR) amplification of the ITS 1 region of ribosomal DNA (rDNA) is combined with a reverse-hybridization line probe assay (LiPA) was used for the identification of six Candida species and four Aspergillus species in pure cultures of clinical isolates, as well as in bronchoalveolar lavage (BAL) fluid samples from 42 patients with various underlying diseases. The results were compared with the results obtained with conventional routine identification methods as well as with a commercial enzyme linked immunosorbent assay (ELISA) galactomannan detection assay and an Aspergillus-specific PCR. No discrepancies between the PCR-LiPA system and routine methods were found for pure cultures of Candida and Aspergillus species except in the case of Aspergillus versicolor. In BAL fluid samples in which Candida species were cultured, the PCR-LiPA system identified more species than did the routine methods. When routine analyses of patient samples were supplemented by adding data obtained by repurifying and re-identifying cultures and by taking isolates obtained from other body sites into account, the results agreed with PCR-LiPA system results in 81% of the cases (34/42). Most of the remaining discrepancies (6/8) involved cases in which such supplementary data were not available. In BAL fluid samples from which A. fumigatus was cultured, the agreement between the PCR-LiPA system and the routine methods was low. Only 2 of 11 BAL samples shown to contain A. fumigatus in ELISA and genus-specific PCR assays were positive in PCR-LiPA system. The PCR-LiPA system enables the simultaneous detection and identification of different fungal species present in pure or mixed populations within 6 h in a single assay. Optimization is required, however, before it is useful as a diagnostic tool in the clinical microbiology laboratory. PMID- 12627808 TI - Recent advances in stem cell neurobiology. AB - 1. Neural stem cells can be cultured from the CNS of different mammalian species at many stages of development. They have an extensive capacity for self-renewal and will proliferate ex vivo in response to mitogenic growth factors or following genetic modification with immortalising oncogenes. Neural stem cells are multipotent since their differentiating progeny will give rise to the principal cellular phenotypes comprising the mature CNS: neurons, astrocytes and oligodendrocytes. 2. Neural stem cells can also be derived from more primitive embryonic stem (ES) cells cultured from the blastocyst. ES cells are considered to be pluripotent since they can give rise to the full cellular spectrum and will, therefore, contribute to all three of the embryonic germ layers: endoderm, mesoderm and ectoderm. However, pluripotent cells have also been derived from germ cells and teratocarcinomas (embryonal carcinomas) and their progeny may also give rise to the multiple cellular phenotypes contributing to the CNS. In a recent development, ES cells have also been isolated and grown from human blastocysts, thus raising the possibility of growing autologous stem cells when combined with nuclear transfer technology. 3. There is now an emerging recognition that the adult mammalian brain, including that of primates and humans, harbours stem cell populations suggesting the existence of a previously unrecognised neural plasticity to the mature CNS, and thereby raising the possibility of promoting endogenous neural reconstruction. 4. Such reports have fuelled expectations for the clinical exploitation of neural stem cells in cell replacement or recruitment strategies for the treatment of a variety of human neurological conditions including Parkinson's disease (PD), Huntington's disease, multiple sclerosis and ischaemic brain injury. Owing to their migratory capacity within the CNS, neural stem cells may also find potential clinical application as cellular vectors for widespread gene delivery and the expression of therapeutic proteins. In this regard, they may be eminently suitable for the correction of genetically-determined CNS disorders and in the management of certain tumors responsive to cytokines. Since large numbers of stem cells can be generated efficiently in culture, they may obviate some of the technical and ethical limitations associated with the use of fresh (primary) embryonic neural tissue in current transplantation strategies. 5. While considerable recent progress has been made in terms of developing new techniques allowing for the long-term culture of human stem cells, the successful clinical application of these cells is presently limited by our understanding of both (i) the intrinsic and extrinsic regulators of stem cell proliferation and (ii) those factors controlling cell lineage determination and differentiation. Although such cells may also provide accessible model systems for studying neural development, progress in the field has been further limited by the lack of suitable markers needed for the identification and selection of cells within proliferating heterogeneous populations of precursor cells. There is a further need to distinguish between the committed fate (defined during normal development) and the potential specification (implying flexibility of fate through manipulation of its environment) of stem cells undergoing differentiation. 6. With these challenges lying ahead, it is the opinion of the authors that stem-cell therapy is likely to remain within the experimental arena for the foreseeable future. In this regard, few (if any) of the in vivo studies employing neural stem cell grafts have shown convincingly that behavioural recovery can be achieved in the various model paradigms. Moreover, issues relating to the quality control of cultured cells and their safety following transplantation have only begun to be addressed. 7. While on the one hand cell biotechnologists have been quick to realise the potential commercial value, human stem cell research and its clinical applications has been the subject of intense ethical and legislative considerations. The present chapter aims to review some recent aspects of stem cell research applicable to developmental neurobiology and the potential applications in clinical neuroscience. PMID- 12627807 TI - CAY-1, a novel antifungal compound from cayenne pepper. AB - CAY-1, a novel saponin from Capsicum frutescens (commercially known as cayenne pepper) was investigated to determine its in vitro antifungal activity, mechanism of action and mammalian cell cytotoxicity. CAY-1 was active against 16 different fungal strains, including Candida spp. and Aspergillus fumigatus [minimum inhibitory concentrations (MIC) ranging from 4 to 16 microg ml(-1)], and was especially active against Cryptococcus neoformans (90% inhibition at 1 microg ml( 1)). Synergistic activity was also observed between CAY-1 and amphotericin B against Candida albicans and A. fumigatus. No significant cytotoxicity was demonstrated when CAY-1 was tested against 55 mammalian cell lines at up to 100 microg ml(-1). Importantly, CAY-1 appears to act by disrupting the membrane integrity of fungal cells. PMID- 12627809 TI - Mapping of the neuronal networks of human cortical brain functions. AB - OBJECTIVE: The principles and methodology of event-related fMRI, electromagnetic source imaging and intracranial evoked potentials will be described along with some examples of the mapping of the neuronal networks of human cortical brain functions with the use of these techniques. INTRODUCTION: Functional brain mapping using PET or fMRI has provided clues on the functioning brain and notably on the functional neuroanatomy of cognitive functions. These mapping possibilities can be used to delineate in an individual patient the brain areas subserving a cerebral function that might be compromised by a surgery in a nearby location, or to target a functional neurosurgical procedure. BACKGROUND: Brain functions and notably "higher brain functions" are served by a complex network of interrelating brain regions. Deeper insights into the functioning of a neuronal network can be gained by adding dynamic, i.e. temporal, information to the functional maps. This will demonstrate the orchestration of the activation of the different brain areas constituting the network, which gives clues to the information processing and therefore to the functioning of the different modules of the network. In order to track the flow of information and the sequential activation of the different brain regions constituting the network, brain activity has to be recorded at the speed of transfer of activation from one neuronal population to the other. The temporal resolution needed to achieve this is not in the range of traditional subtractive or comparative PET or fMRI techniques. NEW DEVELOPMENTS: Novel fMRI methods that record haemodynamic signal changes after single events (event-related fMRI) are now able to determine sequential neural processing by distinguishing the relative onset-time of activity between different areas. The temporal resolution of event-related (ER) fMRI is sufficient to detect changes of mental activity within the order of several hundreds of milliseconds. This allows the exploration of a broad range of cognitive functions. Nevertheless, this technique is currently not rapid enough to observe the transient coordinations and oscillations of neuronal activities occurring across certain cortical areas during the performance of cognitive tasks. The temporal resolution needed for that is within the order of tens or a few milliseconds and is only accessible by EEG or MEG that allow true real-time measurements of the neuronal activity elicited by a stimulus. Surface recordings of multichannel EEG or MEG combined with novel electromagnetic source localisation algorithms allow a relatively precise estimation of the activated areas. A more direct localisation of electric activity is achieved by intracranial recordings in patients having implanted electrodes for diagnostic reasons. In these cases, a high temporal and spatial resolution is achieved but with a limited sampling of brain regions. CONCLUSION: Although the temporal resolution of ER fMRI is due to improve, the temporal measures provided by EEG, MEG or intracranial event-related potentials (ERPs) are absolute, which remains a unique feature of these techniques. Therefore, ER fMRI and electromagnetic source imaging are complementary. The maps obtained with ER fMRI may be refined by electromagnetic ERPs that provide further insights into the temporal coordination or orchestration between the cortical areas already detected by ER fMRI and constituting a neuronal network, and ER fMRI can be used to precisely locate the areas coarsely situated and delineated by electromagnetic source imaging. Thus, the combination of ER fMRI and electromagnetic ERPs is essential in order to produce a mapping method with a millimetre spatial resolution and a millisecond temporal resolution. Future applications should combine these techniques to localise precisely and non-invasively relevant sensory, motor and cognitive processes in order to adequately tailor any brain surgery. PMID- 12627810 TI - Cerebral revascularization. AB - During the last 10 years, there has been a revival of interest in cerebral revascularization procedures. Not only have significant progressions in surgical techniques been published, the use of more advanced diagnostic methods has led to a widening of the indications for cerebral bypass surgery. The purpose of this review is to outline the current techniques for extracranial-to-intracranial (EC/IC) and intracranial-to-intracranial (IC/IC) bypass surgery, as well as to identify the current indications for revascularization procedures based on the available literature. The excimer laser-assisted non-occlusive anastomosis (ELANA) technique is described in more detail because we think that this technique almost completely eliminates the risk of cerebral ischemia due to the temporary vessel occlusion which is currently used in conventional anastomosis techniques. PMID- 12627811 TI - Relative indications for radiosurgery and microsurgery for acoustic schwannoma. AB - The physical and biological principles underlying the use of radiosurgery for the treatment of vestibular schwannomas of up to 2.5 cm in diameter are reviewed together with the historical controversies that have surrounded its introduction. The results in terms of mortality, quality of life, preservation of facial movement and hearing, incidence of shunt-dependent hydrocephalus, cancer neogenesis and brain stem damage are compared in the Marseilles series of 600 microsurgical procedures and 830 Gamma knife procedures and with the peer reviewed literature. The key principles of a steep profile to radiation exposure at the tumour margin, careful topographical planning of the radiation against the tumour shape to minimise the radiation dose to the cranial nerves and brain stem, early tumour swelling, tumour texture and national history of the tumour are analysed. Protocols for the management of unilateral schwannoma, Type II neurofibromatosis (both the Wishart and the Gardner types) and residual/recurrent tumours are presented. In summary, the growth of nearly 97% of vestibular schwannomas (up to 2.5 cm) is arrested by the Gamma knife, the facial nerve is preserved in almost all cases and hearing may be preserved at its pre-operative level in nearly 70% of cases without the complications of microsurgery. PMID- 12627812 TI - The management of brain abscesses. PMID- 12627813 TI - Surgical anatomy of the temporal lobe for epilepsy surgery. AB - Performing temporal lobe epilepsy (TLE) surgery needs secure knowledge of the surgical anatomy. As regards morphological anatomy, the required knowledge includes ability to identify the temporal sulci and gyri with their posterior landmarks, the opercula and borders of the insula on the convexity surfaces, as well as the mesial structures. The anatomical structures delineating the temporal horn have also to be well-recognized by the surgeon from inside the ventricle, namely: the hippocampus with its tiny fimbria bundle, the choroidal fissure and its velum with the attached choroid plexus, fed by the anterior and postero lateral choroidal arteries. As TLE surgery also consists of disconnections, knowledge has to include 1) the (fronto-temporal) uncinate fascicle which is divided by doing limen insulae incision, 2) the (intertemporal) anterior commissure which is laterally interrupted when doing total removal of amygdala and entorhinal cortex, 3) the (fornical) bihippocampal commissure, and more generally the ipsilateral limbic system, which are disconnected when sectionning posteriorly the hippocampal tail and the parahippocampal gyrus, 4) the interhemispheric fibers passing through the corpus callosum via the tapetum when dividing the temporal stem, and 5) many other associative fibers... Functional anatomy has to be perfectly known because the temporal lobe plays a major role, especially in language and memory. Also of paramount importance are the visual and auditory pathways; they are in close relationships with the temporal horn; then they project to the occipital calcarine banks and the temporal operculum, respectively. Surgery in the temporal lobe entails risks of vascular complications; almost all the targets have "dangerous" vascular relationships. Therefore good knowledge in vascular anatomy and regular and intensive training in microsurgery are important prerequisitives for being allowed to perform epilepsy surgery. PMID- 12627814 TI - Clinical and magnetic resonance imaging study of unilateral sideways disc displacements of the temporomandibular joint. AB - The aim of our study was to find symptomatic diagnostic factors for sideways displacement of the temporomandibular joint disc, compared with anterior or rotational disc displacement. A consecutive series of 2310 temporomandibular joints were examined with MRI to define the clinical signs and symptoms particularly related to sideways disc displacement compared to anterior disc displacement with logistic regression model. Bone change of the mandibular condyle and age were negatively related factors in differentiating cases with lateral disc displacement from anterior disc displacement without reduction. Range of mouth opening (over 40 mm) was a significant predictor (odds ratio 4.5865 for lateral disc displacement). This study suggested the wide opening of the mouth to become the predictor for the lateral disc displacement among the cases suspected to have disc displacement without reduction. PMID- 12627815 TI - Changes in response properties of periodontal mechanoreceptors during tooth movement in rats. AB - Using an in vitro model, we investigated the chronological effects of orthodontic force on the response properties of periodontal mechanoreceptors (PMRs) in the rat mandibular first molar (M1). Experimental tooth movement was obtained by attaching a super-elastic titanium-nickel (Ti-Ni) alloy closed coil spring from the mandibular incisors to the right M1. On 1, 2, 3, 4, 7 and 14 days after the appliances were set, three right mandibular molars were extracted and direct stimulation with von Frey hairs was applied to the PMRs remaining in the tooth sockets of right M1. Single unit discharges were recorded from the inferior alveolar nerve. Following results were obtained; (1) in the 1-, 2- and 3-day groups, the mechanical thresholds were significantly lower than those in the control group. In the 4-day group, the mechanical threshold was significantly higher than that of the 3-day group. (2) In the 3-, 4-, 7- and 14-day groups, the conduction velocities of A(beta) units were lower than those in the control group. These results imply that orthodontic force applied to M1 induced functional changes in the PMRs within a few days, suggesting that the PMR seems to respond to orthodontic force at early stage of tooth movement. PMID- 12627816 TI - Sequence of ultrastructural changes of enamel crystals and Streptococcus mutans biofilm in early enamel caries in vitro. AB - Undecalcified mature enamel sections were used to observe the sequence of ultrastructural changes of enamel crystals and Streptococcus mutans biofilm in the early stages of caries. Human enamel blocks were incubated from 1 to 7 days with S. mutans suspension, and the pH of biofilm was measured. They were processed for light microscopic and transmission electron microscopy observations, and the number of bacteria located in the area adjacent to enamel surface counted. It was observed that the pH of the biofilm dropped to 4, after 1 day of incubation and the S. mutans number increased until 4-day. Round shaped enamel crystals were observed in the 2-day specimens and from the 4-day, images of crystals showing defects and perforations were visualized, becoming more defective along the incubation days. The length of time that the enamel was exposed to biofilm was the main factor for enamel crystals demineralization. Current in vitro caries induction system could standardize time-related changes of the property of Streptococcus mutans biofilm and its relation to enamel crystals demineralization at the ultrastructural level, and thus provide a useful model for the evaluation of the effects of various anti-cariogenic agents. PMID- 12627817 TI - Investigation into the usefulness and adverse events of CDDP, 5-fU and dl leucovorin (PFL-therapy) for advanced colorectal cancer. AB - Biochemical modulation of 5-fluorouracil (5-FU) has been verified the evidence of significant improvement of survival and quality of life in patients with advanced colorectal carcinoma. We investigated the therapeutic and adverse drug reaction of intensive chemotherapy using cisplatin (CDDP), 5-FU and dl-leucovorin (LV) (PFL-therapy), which may be producing dual biochemical modulation effect of 5-FU for advanced colorectal carcinoma. Administration schedule was 13 mg/m2 of CDDP, 300 mg/m2 of 5-FU, and 30 mg/body of dl-LV for 5 consecutive days. This regimen was repeated at 3-week intervals in hospital. Sixteen patients were enrolled in this study, most of whom had a history of previous chemotherapy as adjuvant treatment, and the response rate was 25%, with four patients having "partial response" and eight "no change". In respect to performance status, 46% of patients who completed the protocol were markedly improved in spite of their poor performance status before treatment. Moreover, when patients were classified into two groups based on changes of the serum level of CEA, "responder in CEA level" showed better prognosis than "non-responder in CEA level". Major toxicities were nausea, hyperglycemia and neutropenia. Three patients experienced Grade 4 hematological side effect, but these complications resolved quickly in all patients except for one patient. PFL-therapy is effective for advanced colorectal cancer with large tumor burden and showed the same prognostic result as the American and European trials in spite of smaller number of treatment cycles and a history of previous chemotherapy. We will be able to demonstrate the usefulness of this regimen for Japanese patients with advanced colorectal cancers after adding new cases to the present report. PMID- 12627819 TI - Spatial distribution of human respiratory system transfer impedance. AB - Transfer impedance (Ztr) of the respiratory system provides specific information on airways and tissues, but little is known about its spatial distribution in the different thoracoabdominal regions. To study Ztr distribution on the chest wall surface we analyzed five healthy subjects in the supine position by applying a sinusoidal forcing pressure (4, 8, and 12 Hz) at the mouth and measuring airway opening pressure and flow. Three-dimensional positions of 68 reflective markers placed on the chest wall over selected reference points were simultaneously measured by an optoelectronic motion analyzer. A subset of ten points placed on the midline were used to measure chest wall movements in the craniocaudal direction. While the motion of rib cage markers was synchronous, the abdominal markers demonstrated surface waves propagating caudally. The amplitude and phase of these waves were strongly dependent on position and frequency. We used a new method to measure total and local chest wall volume variations to compute the distribution of Ztr over the chest wall. Above 4 Hz we found that Ztr was inhomogeneously distributed and strongly dependent on position and frequency, mainly in the abdomen where the phase was often more more than 180 degrees with high values of modulus. For this reason, we conclude that above 8 Hz Ztr represents rib cage mechanics almost exclusively. PMID- 12627818 TI - Expression of proteoglycan mRNA in human temporomandibular joint disks in temporomandibualr disorders. AB - The purpose of this study was to investigate the expression of PG mRNA in human temporomandibular joint (TMJ ) disks from patients with temporomandibular disorders (TMD). Total RNA was isolated from the TMJ disks. All patients had undergone diskectomy under general anethesia. RT-PCR was performed using primers for the proteoglycans, aggrecan and decorin. On electrophoresis of a lysate of human TMJ disks, decorin was recognized in each sample. In the calcified part of the disk which we recognized visually, aggrecan was also recognized. These results suggest that TMJ disks contain decorin, and are fibrous tissue. If the disk was subjected to excessive compression in addition to tension, cells similar to chondrocytes and expression of aggrecan was induced. Calcification would be initiated in the part of the disk experiencing the increased compression. PMID- 12627820 TI - Reproducibility of image-based computational fluid dynamics models of the human carotid bifurcation. AB - Recent studies have demonstrated the ability of magnetic resonance imaging (MRI) to provide anatomically realistic boundary conditions for computational fluid dynamics (CFD) simulations of arterial hemodynamics. To date, however, little is known about the overall reproducibility of such image-based CFD techniques. Towards this end we used serial black blood and cine phase contrast MRI to reconstruct CFD models of the carotid bifurcations of three subjects with early atherosclerosis, each imaged three times at weekly intervals. The lumen geometry was found to be precise on average to within 0.15 mm or 5%, while measured flow and heart rates varied by less than 10%. Spatial patterns of a variety of wall shear stress (WSS) indices were largely preserved among the three repeat models. Time-averaged WSS was reproduced best, on average to within 5 dyn/cm2 or 37%, followed by WSS spatial gradients, angle gradients, and oscillatory shear index. The intrasubject flow rate variations were found to contribute little to the overall WSS variability. Instead, reproducibility was determined largely by the precision of the lumen boundary extraction from the individual MR images, itself shown to be a function of the image quality and proximity to the geometrically complex bifurcation region. PMID- 12627822 TI - Novel concept for a noninvasive cardiopulmonary monitor for infants: a pair of pajamas with an integrated sensor module. AB - We present a noninvasive cardiopulmonary monitor for use in infants in which sensors are incorporated into conventional pajamas. It consists of dry electrodes for picking up the electrocardiogram and of two capacitive based elastic strain gauges for the measurement of thoracic and abdominal respiratory movements. The quality of the signals was assessed by performing at home seven sleep recordings in parallel with a conventional system and 16 overnight sleep recordings. In the former tests, the acquisition of all mean R-R intervals agreed in both systems with an accuracy of 15.6 ms, determined by the sampling frequency of the commercial system. For the overnight recordings, more than 99% of the R peaks were correctly detected. In 1.5%, both respiratory traces were simultaneously out of range. Finally, it was observed that more saturated episodes and less errors in R-peak detection appeared in prone than in supine position. In summary, these results demonstrated that the dry electrodes can be a good alternative to the sticking electrodes, and that this simple system is reliable. In contrast with the existing monitors, skin irritations are avoided, redundancy of respiratory signals is provided and user-friendliness of the system is reached. PMID- 12627821 TI - Reproducibility study of magnetic resonance image-based computational fluid dynamics prediction of carotid bifurcation flow. AB - The importance of shear stress in the initiation and progression of atherosclerosis has been recognized for some time. A novel way to quantify wall shear stress under physiologically realistic conditions is to combine magnetic resonance imaging (MRI) and computational fluid dynamics. The present study aims to investigate the reproducibility of the simulated flow by using this combined approach. The right carotid bifurcations of eight healthy subjects were scanned twice with MRI within a few weeks. Three-dimensional geometries of the vessels were reconstructed for each scan and each subject. Pulsatile flows through these models were calculated to assess errors associated with the predicted flow parameters. This was done by comparing various wall shear stress indices, including the time-averaged wall shear stress (WSS), oscillating shear index (OSI), WSS Gradients (WSSG) and WSS Angle Deviation (WSSAD). Qualitatively, all the wall shear parameters proved to be highly reproducible. Quantitatively, the reproducibility was over 90% for OSI and WSSAD, but less impressive (60%) for other parameters. Our results indicated that WSS and WSSG values were extremely sensitive to subtle variations in local geometry and mesh design, particularly in regions around the bifurcation apex where WSS values were high and least reproducible. PMID- 12627824 TI - Chronic pulsatile shear stress alters insulin-like growth factor-I (IGF-I) binding protein release in vitro. AB - Insulin-like growth factor-I (IGF-I) is a potent smooth muscle cell mitogen indicated to have a role in vascular disease. IGF-I stimulates proliferation via receptor activation but its activity is mediated by IGF binding proteins (IGFBPs). Since hemodynamics have been linked to vascular proliferative disorders, we studied how pulsatile low (5 +/- 2 dynes/cm2) and high (23 +/- 8 dynes/cm2) shear stresses impacted IGFBP metabolism in bovine aortic endothelial cells using the Cellmax capillary system. We modeled the pulsatile flow in our system using the Womersley model for flow inside a rigid tube and harmonic analysis revealed that the flow was sinusoidal with a frequency of approximately 0.3 Hz for both shear stress treatments. Laminar flow was confirmed and the phase lag between the pressure and the flow found to be insignificant. Thus, our study provides a necessary characterization of this in vitro system as well as an investigation into how shear impacts the IGF axis. We found a significant difference in IGFBP distribution between treatments and, given that IGFBPs regulate IGF-I activity and that IGF-I-independent activities have been suggested for IGFBP-3, suggest that shear stress may indirectly regulate IGF-I activity, and, by extension, the effect of IGF-I on vascular pathologies. PMID- 12627823 TI - Genetically engineered biologically based hemostatic bioassay. AB - Real-time direct measures of hemostatic parameters in vivo are required for optimizing the dynamic delivery of coagulation modifying pharmacotherapies. Typical sensors of physiologic functions in vivo, however, have only a restricted array of sensory inputs, and thus limited capacity to monitor thrombotic and hemostatic activity. To overcome this limitation we have developed a genetically engineered excitable cell line that can be potentially used for an implantable thrombin biosensor. Specifically, we have generated stem cell-derived cardiac myocyte aggregates overexpressing the human thrombin receptor, protease activated receptor-1 (PAR-1), which exploit the inherent electropotential input-output relationship of the cells to detect local changes in thrombin activity. In vitro, the signaling activity of PAR-1 cardiac myocytes was highly responsive to thrombin, inducing a sixfold increase in intracellular cAMP as compared with a twofold increase in control cells. In vivo, the engineered myocytes also detected alterations in local coagulation potential. Specifically, PAR-1 engineered cells implanted in vivo detected local increases in thrombin with a doubling in chronotropic activity compared with a 50% increase in control aggregates. Overall these studies demonstrate the potential of genetic engineering to expand the physiologic signals recognized by excitable cells, and may facilitate the translation of this approach for the real-time monitoring of hemostatic function in vivo. PMID- 12627826 TI - A model of fluid flow in solid tumors. AB - Solid tumors consist of a porous interstitium and a neoplastic vasculature composed of a network of capillaries with highly permeable walls. Blood flows across the vasculature from the arterial entrance point to the venous exit point, and enters the tumor by convective and diffusive extravasation through the permeable capillary walls. In this paper, an integrated theoretical model of the flow through the tumor is developed. The flow through the interstitium is described by Darcy's law for an isotropic porous medium, the flow along the capillaries is described by Poiseuille's law, and the extravasation flux is described by Starling's law involving the pressure on either side of the capillaries. Given the arterial, the venous, and the ambient pressure, the problem is formulated in terms of a coupled system of integral and differential equations for the vascular and interstitial pressures. The overall hydrodynamics is described in terms of hydraulic conductivity coefficients for the arterial and venous flow rates whose functional form provides an explanation for the singular behavior of the vascular resistance observed in experiments. Numerical solutions are computed for an idealized case where the vasculature is modeled as a single tube, and charts of the hydraulic conductivities are presented for a broad range of tissue and capillary wall conductivities. The results in the physiological range of conditions are found to be in good agreement with laboratory observations. It is shown that the assumption of uniform interstitial pressure is not generally appropriate, and predictions of the extravasation rate based on it may carry a significant amount of error. PMID- 12627825 TI - Model-based analysis of matrix metalloproteinase expression under mechanical shear. AB - We report the development of a model-based analysis for identification of the role of transcription factor (TF) binding motifs. A nonlinear mathematical model was formulated to establish the quantitative relationship between the temporal expression profiles and the distribution of known TF binding motifs on regulatory DNA regions. In order to evaluate biological meaning of the nonlinear model, the role of TF binding motifs predicted by the model was examined by a promoter competition assay where specific TF binding motifs were inactivated by a transient transfer of the DNA fragments consisting of the TF binding motifs. Using the shear stress responses of a family of matrix metalloproteinases in human synovial cells as a model system, we showed that the nonlinear formulation was able to approximate the experimentally observed expression profile better than the linear formulation, and the stimulatory and inhibitory roles of TF binding motifs extracted from the model were validated by the competition assay. The results support that an integrated usage of the nonlinear model and the biochemical evaluation assay would contribute to identifying critical regulatory DNA elements in mechanical responses in connective tissue. PMID- 12627827 TI - Magnetic field visualization in applications to pulsed electromagnetic field stimulation of tissues. AB - Electromagnetic field visualization is important in multidisciplinary research on the molecular basis of therapeutic effects of pulsed electromagnetic fields (PEMF). We have compared classic PEMF representations by two-dimensional field lines and field magnitude contour plots with a field representation using three dimensional field isosurfaces. Field simulations were performed for a clinically approved Spinal-Stim Lite system (Orthofix Inc., McKinney, TX). The relatively simple coil system geometry and the predominantly dielectric properties of the surrounding medium (air and human connective tissue) allowed us to develop analytical expressions for the field. The field model was validated by comparison with experimentally measured field values, and with values calculated using a commercial finite-element analysis software package. Two-dimensional field representations by field lines and field contour plots were less intuitive than three-dimensional field isosurface representations to members of the group without an engineering background. Field isosurfaces, represented as three dimensional solids, allowed for direct visualization of PEMF targeting of individual organs (lumbar spine), the extent of the therapeutic field value, and the directional field characteristics. The dynamic characteristic of the field was well illustrated by a sequence of field isosurfaces corresponding to the evolution with time of the electric current waveform (sawtooth) powering the coils. The isosurface representation of the field can be extended to any three dimensional coil system geometry using plotting capabilities of current computer algebra software packages. PMID- 12627830 TI - Self-contained in-the-ear device to deliver altered auditory feedback: applications for stuttering. AB - The design and operating characteristics of the first self-contained in-the-ear device to deliver altered auditory feedback is described for applications with those who stutter. The device incorporates a microdigital signal processor core that reproduces the high fidelity of unaided listening and auditory self monitoring while at the same time delivering altered auditory feedback. Delayed auditory feedback and frequency-altered feedback signals in combination or isolation can be generated to the user in a cosmetically appealing custom in-the canal and completely-in-the-canal design. Programming of the device is achieved through a personal computer, interface, and fitting software. Researchers and clinicians interested in evaluating persons who stutter outside laboratory settings in a natural environment and persons who stutter looking for an alternative or adjunct to traditional therapy options are ideal candidates for this technology. In both instances an inconspicuous ear level alternative to traditional body worn devices with external microphones and earphones is offered. PMID- 12627828 TI - Estimation of musculotendon properties in the human upper limb. AB - The purpose of this study was to develop and apply a general method for estimating the architectural properties of human muscles in vivo. The method consists of a two-phase, nested optimization procedure in which the values of peak isometric force, optimal muscle-fiber length, and tendon slack length are calculated for each musculotendon actuator, knowing muscle volume and the minimum and maximum physiological lengths of the actuator. In phase I, the positions of the bones and the activation levels of the muscles are found by maximizing the isometric torque developed for each degree of freedom at each joint. In phase II, the architectural properties of each musculotendon actuator are found by matching the strength profile of the model to that measured for subjects. The method is used to estimate the architectural properties of 26 major muscle groups crossing the shoulder, elbow, and wrist. Wherever possible, the model calculations are compared against measurements obtained from anatomical studies reported in the literature. Architectural data obtained from our work should be useful to researchers interested in developing musculoskeletal models of the upper limb. PMID- 12627829 TI - Time-dependent entropy estimation of EEG rhythm changes following brain ischemia. AB - Our approach is motivated by the need to generate a rigorous measure of the degree of disorder (or complexity) of the EEG signal in brain injury. Entropy is a method to quantify the order/disorder of a time series. It is the first time that a time-dependent entropy (TDE) is used in the quantification of brain injury level. The TDE was sensitive enough to monitor the significant changes in the subject's brain rhythms during recovery from global ischemic brain injury. Among the different entropy measures, we used Tsallis entropy. This entropy is parametrized and is able to match with the particular properties of EEG, like long-range rhythms, spikes, and bursts. The method was tested in a signal composed of segments of synthetic signals (Gaussian and uniform distributions) and segments of real signals. The real signal segments were extracted from normal EEG, EEG recordings from early recovery, and normal EEG corrupted by simulated spikes and bursts. Adult Wistar rats were subjected to asphyxia-cardiac arrest for 3 and 5 min. The TDE detected the pattern of ischemia-induced EEG alterations and was able to discriminate the different injury levels. Two parameters seem to be good descriptors of the recovery process; the mean entropy and the variance of the estimate followed opposite trends, with the mean entropy decreasing and its variance increasing with injury. PMID- 12627831 TI - Effect of individual Sumai 3 chromosomes on resistance to scab spread within spikes and deoxynivalenol accumulation within kernels in wheat. AB - Two sets of substitution lines were developed by crossing individual monosomic lines of Chinese Spring (recipient) with scab (Fusarium graminearum) resistant cultivar Sumai 3 (donor) and then using the monosomics as the recurrent male parent for four backcrosses (without selfing after each backcross). The disomic substitution lines were separated from selfed BC4F2 plants. Chromosome specific SSR markers were analyzed for polymorphism between Sumai 3 and Chinese Spring. Polymorphic markers were used to identify substitution lines for specific chromosomes. Based on the specific SSR markers, chromosome substitutions occurred in thirty-six lines, and six lines segregated alleles from the two parents or were homozygous for the allele from Chinese Spring. These substitution lines were used to evaluate Type II (spread within the head) and Type V (deoxynivalenol accumulation within kernels) scab resistance. The objective was to use the substitution lines to evaluate the effect of individual chromosomes of Sumai 3 on Type 11 and Type V scab resistance in the greenhouse. Significant differences in Type II scab resistance and deoxynivalenol (DON) levels among different Chinese Spring (Sumai 3) substitution lines were detected. Positive chromosome substitution effects on Type II scab resistance were found on chromosomes 2B, 3B. 6B, and 7A from Sumai 3. Chromosomes 3B and 7A also reduced DON accumulation within the kernels, while chromosomes IB, 2D, and 4D from Sumai 3 increased DON concentration. Chromosome 7A from Sumai 3 had the largest effect on resistance to scab spread and DON accumulation. Additional research is in progress on the scab resistance conferred by chromosome 7A. PMID- 12627832 TI - Variable patterns of chromosome synapsis at pachytene in Hordeum vulgare x H. bulbosum hybrids and their parents. AB - Synaptonemal complexes (SC) have been analysed in barley (Hordeum vulgare), H. bulbosum and two F, hybrids between them. These hybrids show different recombination frequencies and at pachytene show significant differences in the total length of SC formed and in the extent of synapsis. Higher recombination frequency in the hybrids was correlated with a greater total SC length. Differences in SC length were also observed between the parental species with H. bulbosum having a greater SC length than H. vulgare. However, species and hybrid can have similar SC lengths but clearly different recombination frequencies and, therefore, the relationship between SC length and recombination is not clear-cut. PMID- 12627833 TI - Inheritance of dense spike in diploid wheat and Aegilops squarrosa. AB - The individuals of diploid wheat Triticum boeoticum, T. monococcum and T. sinskajae and goatgrass Aegilops squarrosa were picked out with screening the dense spike characteristics. The dense-spike accessions were discovered in diploid wheat (T. sinskajae) and Ae. squarrosa. Inheritance of the dense spike was studied. The trait was found to be controlled by a recessive gene in T. sinskajae and by an incomplete dominant gene in Ae. squarrosa. The dosage effect of dominant gene C was detected in interspecific pentaploid F1 hybrid plants T. compactum x T. palmovae (2n =35, A(u)A(b)BDD genome). The spike of pentaploid hybrid was not so dense as compared to hexaploid wheat T. compactum. This is the first report showing similarity of the expression of dominant gene C on D genome of the hexaploid wheat to that of dense spike gene in Ae. squarrosa. The existence of dense-spike accessions of Ae. squarrosa allows us to hypothesize that the origin of T. compactum is independent from that of common wheat. PMID- 12627834 TI - Chromosome analysis using different staining techniques and fluorescent in situ hybridization in Cerithium vulgatum (Gastropoda: Cerithiidae). AB - In the present paper one population of the large" subtidal mollusc Cerithium vulgatum Bruguiere, 1792 (Gastropoda: Cerithiidae) from the Northwestern coast of Sicily was investigated from a karyological point of view. The chromosome complement was Giemsa stained, conventionally karyotyped in 18 homomorphic chromosome pairs (10 bi-armed and 8 mono-armed), and subsequently analysed using silver, CMA3 and DAPI staining, and fluorescent in situ hybridization (FISH) with three repetitive DNA probes [ribosomal DNA (rDNA), (TTAGGG)n and (GATA)n]. FISH with the rDNA probe consistently mapped major ribosomal sites (18S-28S rDNA) in the terminal region of the short arms of one small sized mono-armed chromosome pair. Ribosomal DNA was transciptionally active as indicated by its preferential impregnation with silver nitrate (Ag-NOR) and did not contain a high amount of GC base pairs as suggested by the lack of a bright CMA3 fluorescence. The (TTAGGG)n telomeric probe was hybridized to the termini of nearly all chromosomes, thus demonstrating that, in C. tulgatum, this sequence has been conserved during the genomic evolution. The finding of the telomeric hexanucleotide in six species belonging to the three high taxa of Gastropoda supports the notion that this sequence is widespread within this class. The (GATA)n probe did not label any chromosome regions except for a minute terminal area of a single bivalent at pachytene stage. PMID- 12627835 TI - Multiple Nors in Bryconamericus aff. exodon (Osteichthyes, Characidae, Tetragonopterinae). AB - Fifteen examples of Bryconamericus aff. exodon from the Tres Bocas stream in the basin of the Tibagi river (Parana, Brazil) were analyzed. They presented a diploid number of 52 chromosomes. Multiple NORS were detected by silver nitrate impregnation in the telomeric region on the short or long arm of different chromosomes, showing a variation from 2 to 5 NOR-bearing chromosomes. Treatment with chromomycin A3 (CMA3) showed regions rich in GC in different chromosomes, probably coincident with the AgNORS. In situ hybridization by fluorescence (FISH) with 18S rDNA probe showed 8 hybridization signs in the telomeric regions of the chromosomes, proving that this B. aff. exodon population has a greater number of ribosomal cistrons than were detected by Ag-banding. PMID- 12627836 TI - Bromus fasciculatus presl--a third diploid progenitor of Bromus section Genea allopolyploids (Poaceae). AB - Allozyme variation of ten heterozymes of seven enzymes among five accessions of a rare diploid Bromus fasciculatus was analysed with the use of PAGE and compared with that for six other species of section Genea of the genus Bromus. Allozymes charasteristic for diploids B. tectorum and B. fasciculatus are combined in fixed heterozygous phenotypes of tetraploid B. rubens. Fixed heterozygous phenotypes of tetraploid B. madritensis combine one allozyme of B. fasciculatus with another of diploid B. sterilis at each of the loci studied. Of the three diploids studied, only B. sterilis fits well for a role of a genome donor for the polyploid B. diandrus-rigidus complex. Bromus fasciculatus thus appears to be a diploid ancestor for the two tetraploids of section Genea, B. rubens and B. madritensis. PMID- 12627837 TI - Interspecific polymorphism at non-coding regions of chloroplast, mitochondrial DNA and rRNA IGS region in Elymus species. AB - Several published universal primers for amplification of non-coding regions of chloroplast, mitochondrial and ribosomal (rRNA) IGS region were tested whether they can amplify respective regions in Elymus species. PCR-RFLP analysis of the chloroplast, mitochondral DNA, and rRNA IGS region of the genus Elymus was used to determine if the method could be employed to detect inter-specific variation in this genus. Published universal primers for amplification of trnK [tRNA-Lys (UUU) exon 1]-trnK [tRNA-Lys (UUU) exon2], and mitochondrial nad1 exon B-nadl exon C intron successfully amplified the respective regions in Elymus species. However, the primers for amplification of chloroplast trnD-trnT intron and rRNA IGS failed to amplify the respective region in Elymus species. New primer pairs were designed and successfully amplified the cpDNA trnD-trnT intron and rRNA IGS region in Elymus species. The amplification products were digested with seven restriction enzymes. The results showed that the investigated regions of chloroplast and mitochondrial genomes are variable in most of the tested taxa and contain multiple variable regions. These regions should serve as useful molecular markers in phylogenetic studies of closely related species, at least at the interspecific level in Elymus. It is likely that further studies, including larger sample sizes, more regions of these genomes and/or more powerful methods for the detection of cpDNA and mt DNA variation will reveal additional variation for this genus. Highly inter- and intra-specific polymorphisms for rRNA IGS region were detected, suggesting the IGS will be a useful molecular marker for population studies of Elymus species. PMID- 12627838 TI - Meiotic studies in Dysdercus Guerin Meneville, 1831 (Heteroptera: Pyrrhocoridae). II. Evidence on variations of the diffuse stage between wild and laboratory inbred populations of Dysdercus chaquensis Freiberg, 1948. AB - Dysdercus Guerin Meneville, 1831 comprises insect species that are often serious pests of cotton both in the Old and New World, representing the only taxon from Pyrrhocoridae in the Neotropical Region. The genus is cytologically characterized by possession of holokinetic chromosomes and a prereductional type of meiosis. So far, only seven species from the Old World and five species from the Neotropical Region have been cytogenetically described. In the present report we compare the male meiosis from both natural and inbred populations of Dysdercus chaquensis Freiberg, 1948. Our results demonstrated that even though both populations share the same diploid chromosome number, the presence of a diffuse stage was found to be committed to the wild population of the species. Furthermore, the diffuse stage was found in a high frequency in all analysed wild specimens. indicating the long duration of this period among the regular meiosis of D. chaquensis. Taking into account that the diffuse stage is connected with an intense and long period of cellular growth, and with an important transcriptional activity, the absence of this stage in all the inbred specimens of D. chaquensis could be related with the lack of unfavourable physiological conditions due to the environmental uniformity along seven years of inbreeding. PMID- 12627839 TI - Permanent translocation heterozygosity in dioecious Baccharis coridifolia DC. (Asteraceae). AB - The karyotype of Baccharis coridifolia DC., a dioecious diploid species consists of 16 chromosomes with median (m) and 2 with submedian centromeric positions (sm) (2n = 18). Meiotic analyses of 219 male plants belonging to 12 populations showed across a wide geographical range one quadrivalent and seven bivalents in all of them. The quadrivalent involves the four biggest m chromosomes and probably constitutes a sex multivalent carrying in tight linkage the sex genes and provides a mechanism whereby permanent translocation heterozygosity is maintained. In metaphase I alternate orientation of the quadrivalent was predominant (85.71%). Fourteen megaspore mother cells were analyzed and did not show the quadrivalent. A model for the origin and maintenance of the sexual translocation system is presented. It is assumed that there are two non-allelic factors M and n; M would be a dominant female suppresor and n a recessive male suppresor. The excess of male plants observed in several populations at different localities might result from heterotic effects of some gene complexes carried by the translocation. This excess probably ensures a good supply of pollen for the vagaries of insect pollination at flowering time. PMID- 12627840 TI - Reproduction mode of an artificial allotetraploid carp (Pisces; Cyprinidae). AB - Mature eggs of allotetraploid carp were activated by inactive sperm or crossed with normal sperms of common carp (Cyprinus carpio), crucian carp (Carassius auratus), Chinese blunt snout bream (Megalobrama amblycephala), Hemiculter leucisculus and Pseudorasbora parva. Chromosome counts showed that all offspring of these crosses presented a mode number of 200 chromosomes (4n = 200), and their morphological traits are much like maternal. Microsatelite marker and RAPD patterns between allotetraploid maternal and its offspring, reproduced from different paternal species, were identical. Cytological, morphological and molecular evidences suggested that allotetraploid carp female nucleus would not fuse with any male nucleus and its reproduction mode might be gynogenesis and therefore their offspring are retaining their tetraploidy and give origin to clonal individuals. PMID- 12627841 TI - New C-band protocol by heat denaturation in the presence of formamide. AB - C-banding techniques detect the presence of constitutive heterochromatin, which is usually located in centromeric regions of chromosomes in the majority of analysed species. The common method for C-banding used over the last 30 years involves treatment with a mild alkali barium hydroxide 5% Ba(OH)2 at 50 degrees C for 5-15 min and subsequent incubation in salt solution (2 x SSC at 60 degrees C for 1 h). We here present a new, easy and reliable technique for C-banding, which basically involves heat denaturation of chromosomal DNA in the presence of formamide and incubation in 2 x SSC at room temperature. PMID- 12627842 TI - Genetic relationships among ten endod types as revealed by a combination of morphological, RAPD and AFLP markers. AB - The genetic relationships among ten types of endod (Phytolacca dodecandra) cultivated by the Institute of Pathobiology of the Addis Ababa University to combat the disease bilharzia in Ethiopia were studied using morphology and molecular markers. A total of 18 morphological characters, 194 amplified fragment length polymorphism (AFLP) and 42 random amplified polymorphic DNA (RAPD) markers were used to determine genetic proximity between types. Genetic distance and cluster analysis of the AFLP data revealed the lack of genetic difference between E47 and E48 but relatively wider genetic difference among the other endod types. Cluster and principal component analyses performed on the AFLP and RAPD markers demonstrated the presence of distinct separation of E56 but not that of E44 from the others. The AFLP and RAPD data, thcrefore, did not support the hypothesis that the superiority of E44 in agronomic traits and molluscicidal potency is linked to its distinct genetic difference from the other endod types. Matrices correspondence tests demonstrated the presence of greater correspondence between AFLP and RAPD data (r = 0.842) but not between the morphology and that of AFLP and RAPD. This indicates the correspondence more between the two DNA markers systems than either of them with morphological traits. The cophenetic correlation coefficients also revealed poor fit for morphology (r = 0.716), good fit for RAPD (r = 0.872) and very good fit for AFLP (r = 0.975), reflecting the hyper variability and higher resolving power of AFLP. PMID- 12627843 TI - Isolation and cross-species amplification of microsatellite loci in the Siberian jay (Perisoreus infaustus). PMID- 12627844 TI - Molecular genetics of paroxysmal nocturnal hemoglobinuria. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hematopoietic stem cell disorder characterized by the clonal expansion of glycosylphosphatidylinositol (GPI)-deficient cells that leads to complement-mediated hemolysis. A somatic mutation in the PIG-A gene involved in GPI biosynthesis causes a deficiency of GPI-anchored proteins. However, it is evident that the clonal expansion of GPI deficient cells is not caused by only the PIG-A mutation and that other changes should be involved in the development of PNH. Some patients with aplastic anemia (AA) develop PNH. Furthermore, it has been reported that most patients with AA and refractory anemia (RA) who carry HLA-DRB1*15 and show a good response to immunosuppressive therapies have an expanded population of GPI-deficient clones. This finding, together with recent data showing resistance of GPI-deficient cells to cytotoxic cells, suggests that GPI-deficient cells escape immunologic attack and are positively selected in the autoimmune environment. However, GPI-deficient clones found in AA and RA are generally small and do not increase to near complete dominance. Therefore, 1 or more additional genetic abnormalities that confer the growth phenotype on GPI-deficient cells are probably required for fully developed PNH or so-called florid PNH. The next 10 years should witness the discovery of the molecular mechanisms of immunologic selection and the identification of abnormalities involved in the further clonal expansion of PNH cells. PMID- 12627845 TI - Clinical manifestations of paroxysmal nocturnal hemoglobinuria: present state and future problems. AB - The clinical pathology of paroxysmal nocturnal hemoglobinuria (PNH) involves 3 complications: hemolytic anemia, thrombosis, and hematopoietic deficiency. The first 2 are clearly the result of the cellular defect in PNH, the lack of proteins anchored to the membrane by the glycosylphosphatidylinositol anchor. The hemolytic anemia results in syndromes primarily related to the fact that the hemolysis is extracellular. Thrombosis is most significant in veins within the abdomen, although a number of other thrombotic syndromes have been described. The hematopoietic deficiency may be the same as that in aplastic anemia, a closely related disorder, and may not be due to the primary biochemical defect. The relationship to aplastic anemia suggests a nomenclature that emphasizes the predominant clinical manifestations in a patient. This relationship does not explain cases that appear to be related to myelodysplastic syndromes or the transition of some cases of PNH to leukemia. Treatment, except for bone marrow transplantation, remains noncurative and in need of improvement. PMID- 12627846 TI - Pathogenesis of selective expansion of PNH clones. AB - Hemolysis, a characteristic of paroxysmal nocturnal hemoglobinuria (PNH), is caused by the expansion of an affected stem cell with a mutation of the PIG-A gene. Increasing evidence has shown that the presence of the PIG-A mutation alone does not induce the expansion. Two theories have been proposed. One, the growth advantage hypothesis, is supported by current data indicating the presence of several intrinsic alterations that might confer a proliferative advantage to PNH clones over normal cells. Alternatively, the PIG-A mutation might confer a relative survival advantage to PNH clones. This theory is supported by clinical observation indicating that PIG-A mutant cells survive immune-mediated bone marrow injury in patients with aplastic anemia, PNH, and myelodysplastic syndromes. The latter theory is also supported by current experimental data indicating that PIG-A mutant cells are relatively resistant to cytotoxic attack by natural killer cells and cytotoxic T-lymphocytes. The 2 theories appear complementary rather than mutually exclusive. Rapid progress in this field can be expected in the near future. PMID- 12627847 TI - Management issues in paroxysmal nocturnal hemoglobinuria. AB - Paroxysmal nocturnal hemoglobinuria (PNH) arises in the setting of bone marrow injury. Thus, management decisions must take into account whether symptoms are a consequence of the underlying marrow failure or of the expansion of the clone of the PIG-A mutant hematopoietic cells. The primary clinical manifestations of PNH are intravascular hemolysis and thrombophilia. Currently available options for treatment of the hemolysis of PNH are unsatisfactory, but the recent development of specific inhibitors of complement for use in treating human disease should make possible effective management of this pathology. The fundamental basis of the thrombophilia of PNH has not been elucidated. Currently, empiric anticoagulant therapy is the foundation for treating the thromboembolic complications of PNH. The role of warfarin prophylaxis, however, remains an area of active debate. Pregnancy in a patient with PNH presents special concerns about fetal/maternal well-being because of the high potential for thromboembolic complications. Bone marrow transplantation can be considered curative, but the decision to recommend this treatment must take into account factors related both to PNH and to comorbid conditions. Refining the technology for both gene therapy (by transducing stem cells with a functional PIG-A gene) and autotransplantation (by using stem cells selected for the expression of glycosyl phosphatidylinositol anchored proteins) remain challenges for the future. PMID- 12627848 TI - Proliferation of human progenitor cells in a long-term culture system is more efficiently sustained by the addition of Flt-3 ligand or megakaryocyte growth and development factor than by Kit ligand. AB - INTRODUCTION: We compared the effects of the early-acting growth factors (GF), Flt-3 ligand (FL), c-Kit ligand (KL), and leukemia inhibitory factor (LIF), and the late-acting GF, granulocyte-colony stimulating factor (G-CSF) and megakaryocyte growth and development factor (MGDF), added alone in human long term marrow culture (LTMC). MATERIALS AND METHODS: The GF were used in primary cultures of mononuclear cells (MNC) and in cocultures of CD34+ cells on murine preestablished MS-5 stromal layers. GF activity was assessed as nonadherent and adherent progenitor cell production and cobblestone area formation at week 5. RESULTS: In this system, only FL, KL, and MGDF significantly stimulated early stages of hematopoiesis, whereas only G-CSF stimulated the proliferation of mature progenitor cells within the granulo-monocyte lineage and no effect was observed with LIF. FL displayed the strongest activity, and MGDF was more efficient than KL, both in primary cultures of MNC and in cocultures of CD34+ cells. However, the stimulatory effects of these GF used alone were dependent on the presence of a stromal layer. CONCLUSION: These LTMC data emphasize the particular roles for FL and MGDF in the stimulation of primitive hematopoiesis. PMID- 12627849 TI - Granulocyte colony-stimulating factor-induced terminal maturation of human myeloid cells is specifically associated with up-regulation of receptor-mediated function and CD10 expression. AB - The acute promyelocytic leukemia cell line NB4 was differentiated by all-trans retinoic acid (ATRA), which enhanced the superoxide-producing capacity stimulated by the chemotactic peptide and phorbol ester in this cell line. Granulocyte colony-stimulating factor (G-CSF) by itself had no effect on NB4 cells but exerted additional enhancing effects on the respiratory burst activity in the presence of ATRA. This finding was not due to the induction of G-CSF receptor by ATRA, because NB4 cells expressed abundant G-CSF receptor with or without ATRA. Unlike ATRA, G-CSF enhanced superoxide release stimulated by the chemotactic peptide but not by phorbol ester. In addition, G-CSF but not ATRA attenuated cell death and enhanced survival during differentiation. Cell surface expression of the chemotactic peptide receptors CD33 and CD10 but not of CD11b and CD11c was up regulated by ATRA plus G-CSF far more profoundly than by ATRA alone. Fundamentally identical but slightly different phenomena for the cell surface expression of CD33 and CD10 were observed in the normal human bone marrow mononuclear cells; G-CSF induced CD10 even in the absence of ATRA and down regulated CD33 in normal cells. The present results indicate that G-CSF-induced terminal maturation of human myeloid cells is associated with up-regulation of receptor-mediated function and CD10 expression. PMID- 12627850 TI - Multicenter prospective study of clonal complications in adult aplastic anemia patients following recombinant human granulocyte colony-stimulating factor (lenograstim) administration. AB - To elucidate the relationship between treatment with granulocyte colony stimulating factor (G-CSF) and the development of chromosomal abnormalities and clonal evolution in adult aplastic anemia (AA) patients, we performed a prospective multicenter study. Of the 104 registered patients, 91 were found by the central review committee to have the diagnosis of AA. Of the 91 patients, 84 were determined to be evaluable in this study and were treated with regimens including G-CSF (lenograstim). A time-course study (at registration and 6 and 12 months after registration) of G-banded chromosomes, fluorescence in situ hybridization (FISH) analysis for monosomy 7, and morphological assessment of bone marrow was performed with these patients during the 1-year follow-up period. G-banding analysis demonstrated the development of cytogenetic abnormalities in 10 (16.1%) of the 62 evaluable patients. The most common aberration was monosomy 7. FISH analysis demonstrated monosomy 7 in 7 (10.4%) of the 67 evaluable patients. Evolution into myelodysplastic syndrome (MDS) was observed in 5 (7.7%) of the 65 patients who underwent morphological assessment of bone marrow. All patients who developed cytogenetic abnormalities or MDS received immunosuppressive agents and/or steroids with G-CSF. This study demonstrated that some adult AA patients exhibit evolution into MDS and development of chromosomal abnormalities such as monosomy 7. Although the incidence seems to be comparable with that found in previous studies, further long-term follow-up will be necessary to confirm the relationship between G-CSF and the development of chromosomal abnormalities and MDS. PMID- 12627851 TI - Prevalence of intravascular large B-cell lymphoma with bone marrow involvement at initial presentation. AB - We used immunohistochemical analysis to investigate 146 bone marrow (BM) clot specimens from patients with fever of unknown origin (FUO) (n = 124) or hemophagocytic syndrome (HPS) (n = 22). Intravascular lymphoma (IVL) primarily involving BM was detected in 12 (8.2%) of the 146 patients. Diagnosis was based on the presence of CD20+ and CD79a+ tumor cells confined within the lumina of sinuses and surrounded by CD34+ endothelial cells. Of the 12 IVL cases, 6 were CD5+; of these 6 CD5+ cases, 5 were positive for vimentin. The finding of a considerably high prevalence of IVL tumor cells in BM from patients with FUO or HPS suggests that immunohistochemical examination of BM may be helpful in the diagnosis of IVL in these patients. Vimentin coexpression in CD5+ IVL might be evidence of origin from a subset of prefollicular B-cells. PMID- 12627852 TI - Deoxycoformycin-containing combination chemotherapy for adult T-cell leukemia lymphoma: Japan Clinical Oncology Group Study (JCOG9109). AB - Aggressive adult T-cell leukemia-lymphoma (ATL) generally has a very poor prognosis. Deoxycoformycin (DCF, pentostatin), an inhibitor of adenosine deaminase, has shown promising therapeutic efficacy for ATL. To develop a new effective therapy against aggressive ATL, we carried out a multicenter phase II study of DCF-containing combination chemotherapy. Sixty-two previously untreated patients with ATL (34, 21, and 7 patients with diseases of the acute, lymphoma, and unfavorable chronic types, respectively) were enrolled, but 2 were ineligible because they were judged to be favorable chronic types. A regimen of 1 mg/m2 vincristine intravenously on days 1 and 8, 40 mg/m2 doxorubicin intravenously on day 1, 100 mg/m2 etoposide intravenously on days 1 through 3, 40 mg/m2 prednisolone orally on days 1 and 2, and 5 mg/m2 DCF intravenously on days 8, 15, and 22 was administered every 28 days for 10 cycles unless disease progression or toxic complications occurred. Fifty-two percent of 60 eligible patients responded (95% confidence interval [CI], 38%-65%), with 17 patients (28%) achieving a complete response (CR) (95% CI, 17%-41%) and 14 achieving a partial response. The CR rate was inferior to those of both the previous Japan Clinical Oncology Group (JCOG) study (JCOG8701, 43%), a 9-drug combination chemotherapy of the second generation, and the subsequent JCOG9303 study (35%), a granulocyte colony stimulating factor-supported, dose-intensified, 9-drug regimen. The median survival time of the 60 eligible patients in JCOG9109 was 7.4 months, and the estimated 2-year survival rate was 15.5%; these results were identical with those of JCOG8701 but inferior to those of JCOG9303. Grade 4 neutropenia and infection of grade 3 or greater were frequent (67% and 22%, respectively), and treatment related death was observed in 4 patients (7%), septicemia in 2, and cytomegalovirus pneumonia in 2. We conclude that DCF-containing combination chemotherapy is not a promising regimen against aggressive ATL. PMID- 12627853 TI - Pyomyositis as a focus of infection in hematological disorders: a report of 3 cases. AB - The cases of 3 patients with pyomyositis associated with hematological disorders are reported. A 40-year-old man in the blastic phase of chronic myelogenous leukemia and 2 men aged 46 and 71 years with neutropenia due to myelodysplastic syndromes all reported high fever and severe local myalgia and had marked elevation of C-reactive protein. Magnetic resonance imaging revealed muscle abscesses or fasciitis, and the findings led to the diagnosis of pyomyositis. Methicillin-resistant Staphylococcus aureus was isolated from the abscesses of 2 patients, and surgical drainage proved more effective than did antimicrobial agents. It should be recognized that pyomyositis is a possible source of infection in patients with hematological disorders. PMID- 12627854 TI - Generation of CD3+ CD56+ cytokine-induced killer cells and their in vitro cytotoxicity against pediatric cancer cells. AB - A certain number of pediatric cancer patients still succumb to relapse following conventional treatment of their malignancies. One of the mechanisms of relapse is escape from immunity. Adoptive cellular immunotherapy with effector cells has the potential to overcome this escape. In adults, the CD3+ CD56+ cell, a cytokine induced killer (CIK) cell, appears to be a promising effector cell type with the greatest cytotoxicity. This effector cell type may work in children as well. No similar studies with children have been published. We speculated that expanded CD3+ CD56+ cells obtained from pediatric cancer patients during remission would act similarly against various pediatric tumor cell lines; therefore, we undertook the present study to find support for our speculation. This study was undertaken to generate and expand CD3+ CD56+ CIK cells from normal peripheral blood mononuclear cells (PBL) obtained from 6 children with cancer (2 with acute lymphoblastic leukemia, 2 with large cell lymphoma, and 2 with osteosarcoma) in remission after intensive chemotherapy and to study the cytotoxic activities of these cells against chronic myeloid leukemia cell line K562 t(9;22), 4 pediatric tumor cell lines [infant acute lymphoblastic leukemia RS4 t(4;11), TEL/AML acute lymphoblastic leukemia REH t(12;21), alveolar rhabdomyosarcoma Rh-Cr t(2;13), and Ewing sarcoma EW-Le t(11;22)], and 2 pediatric glioblastoma multiforme cultured cell lines (G74 and G77). CIK cells were generated and expanded in culture medium to which interferon gamma, monoclonal antibody against CD3, and interleukin 2 were added at appropriate times. Cells were counted by flow cytometry. Net lactate dehydrogenase release from target cells incubated with CIK cells was used as an index of CIK cell cytotoxicity against various pediatric tumor cell lines. The results show that after 21 days in culture CD3+ CD56+ CIK cells derived from the 6 pediatric patients accounted for a median of 28.3% of the entire culture (range, 10.7%-36.4%). Before expansion no such cells were found in any of the 6 children. Median lytic activity rates of CIK cells were 45.5% to 64.5%, rates that contrasted drastically to the lytic activity rates of PBL, which were only 8% to 12%. The findings of the present study are encouraging. They provide information for developing adoptive immunotherapy for future clinical trials with pediatric cancer patients, particularly those patients with minimal residual disease after intensive chemotherapy or stem cell transplantation (especially nonmyeloablative transplantation procedures). PMID- 12627855 TI - Clinical significance of fragmented red cells after allogeneic bone marrow transplantation. AB - To clarify the clinical significance of the presence of fragmented red cells (FRC) after allogeneic bone marrow transplantation (BMT), we measured the incidence and degree of FRC and their relationships to clinical features. The percentages of FRC (%FRC) were measured in 50 patients on weeks -2, 0, 2, 4, 6, 8, 10, and 12. The %FRC in pre-BMT patients (mean, 0.52%; range, 0.04%-1.56%) was higher than in healthy control subjects (mean, 0.08%; range, 0.02%-0.27%). The highest %FRC (> or = 1.3%) were seen in 2 pre-BMT and 17 post-BMT patients. Eight patients who developed thrombotic microangiopathy (TMA) showed %FRC values that were significantly higher than those in patients without TMA. However, the timing of elevated %FRC was delayed until several days after the onset of intravascular hemolysis and/or a drop in platelet count. Of the patients who did not experience TMA, 5 patients with infection and 4 patients with acute graft-versus-host disease (GVHD) also showed significant elevation of %FRC during the clinical course. Furthermore, multivariate analysis results demonstrated that TMA and infection had a statistically significant effect on the high value of %FRC. These findings indicate that the appearance of FRC is a common phenomenon in patients undergoing BMT and is not a predictive factor for the early diagnosis of TMA, although FRC is one of the main laboratory findings in TMA. Furthermore, an increased %FRC is seen in other post-BMT clinical settings, such as infection and acute GVHD. PMID- 12627856 TI - Heparin-induced thrombocytopenia: acknowledging its presence in low-molecular weight heparin therapy. AB - Low-molecular weight heparin (LMWH) is associated with a lower incidence of heparin-induced thrombocytopenia (HIT) than is unfractionated heparin. We describe a 75-year-old woman who developed HIT with thrombotic manifestations following the use of nadroparin calcium. Subsequent anticoagulation was achieved with warfarin. This case serves to highlight an important complication that cannot be ignored despite its low incidence. The majority of HIT cases are likely to occur in LMWH-treated patients because LMWH replaces unfractionated heparin for most indications. The lack of suitable alternative anticoagulant treatments for patients with HIT in Singapore is also emphasized. PMID- 12627857 TI - Platelet recovery in patients with idiopathic thrombocytopenic purpura after eradication of Helicobacter pylori. AB - The relationship between Helicobacter pylori infection and idiopathic thrombocytopenic purpura (ITP) has been investigated in several studies. We investigated the prevalence of H. pylori infection and the clinical effects of eradication in 22 Japanese patients with chronic ITP. H. pylori infection was found in 14 (63.6%) of the patients by histologic and culture examinations of biopsy samples obtained by gastrointestinal endoscopy. H. pylori was eradicated by proton pump inhibitors and 2 kinds of antibiotics in 13 (92.9%) of the 14 patients in whom the results of treatment could be evaluated. Five (38.4%) of those 13 patients had platelet recovery (platelet count of more than 100 x 10(9)/L and an increase of more than 30 x 10(9)/L with respect to the baseline value) after eradication. The median follow-up period was 15 months. One patient who had a complete response had a partial relapse after cessation of prednisolone treatment without any evidence of H. pylori reinfection. Another patient, in whom H. pylori was not eradicated even after 2 treatment sessions, had a partial response after treatment. A screening examination for H. pylori infection may be necessary for Japanese patients with newly diagnosed ITP. Although the exact mechanism underlying platelet recovery after H. pylori eradication is not clear, the results of this study indicated that H. pylori eradication treatment is a good option for some patients with chronic ITP. PMID- 12627858 TI - International comparison of research productivity in hematology. PMID- 12627859 TI - Pharmacokinetics of a standard dose of cytarabine in a patient with acute promyelocytic leukemia undergoing continuous ambulatory peritoneal dialysis. PMID- 12627861 TI - Gray platelet syndrome presenting as menorrhagia. PMID- 12627860 TI - Kawasaki disease in a patient with Wiskott-Aldrich syndrome: an increase in the platelet count. PMID- 12627862 TI - Meeting report: Tenth International Conference on the Cell and Molecular Biology of Chlamydomonas. PMID- 12627863 TI - Meeting report: XIVth meeting of the International Society for Evolutionary Protistology, Vancouver, Canada, June 19-24, 2002. PMID- 12627864 TI - Meeting report: Culture Collections of Algae: Increasing Accessibility and Exploring Algal Biodiversity. An international meeting at the Sammlung von Algenkulturen at Gottingen University (SAG), Germany; September 2-6, 2002. PMID- 12627865 TI - Expanding the molecular toolkit for Chlamydomonas reinhardtii--from history to new frontiers. PMID- 12627866 TI - Morphological, ecological and molecular studies of Vannella simplex Wohlfarth Bottermann 1960 (Lobosea, Gymnamoebia), with a new diagnosis of this species. AB - Vannella simplex (Gymnamoebia, Vannellidae) is one of the most common amoebae species, recorded from a variety of regions. It was originally described as a freshwater species, but has also been reported from shallow-water regions of the Baltic Sea. In the present work, we investigated the morphology and biology of three V. simplex isolates, originating from geographically distant regions. Among them is one brackish water strain, isolated from artificial cyanobacterial mats, which were originally sampled in Niva Bay (Baltic Sea, The Sound). The strain is cyst-forming and can thrive at salinity ranges from 0-50 ppt. Phylogenetic relationships were investigated by sequencing partial SSU rDNA of the cultured V. simplex isolates. Additional sequences were obtained from four environmental DNA extractions of sediment samples collected from different localities in Switzerland. Analysis of all obtained sequences revealed a monophyletic group. Based on the analysis and comparison of morphological, ecological and molecular data sets we compiled a distribution map of V. simplex and propose an emendation of this species. PMID- 12627867 TI - The phylogeny of Goussia and Choleoeimeria (Apicomplexa; Eimeriorina) and the evolution of excystation structures in coccidia. AB - The phylogenetic relationships of Goussia janae and Choleoeimeria sp. were analyzed using the small subunit ribosomal RNA gene (SSU rDNA). This is a first attempt to study the molecular phylogeny of coccidian genera parasitizing strictly poikilotherm hosts. The biliary Eimeria-like coccidia of reptiles classified into the genus Choleoeimeria form a sister clade to the family Eimeriidae, which confirms the separate generic status of the genus Choleoeimeria. The position of Goussia is less robustly resolved, since it forms a trichotomy with the Eimeriidae and Sarcocystidae, or alternatively constitutes the earliest branch of the coccidian lineage. Morphological similarities, namely the extracytoplasmic location of the endogenous stages, and the presence of sutures in the sporocyst wall are discussed in the context of the traditional classification of eimeriids. In contrast to the morphology-based systematics, the monophyly of Goussia and Choleoeimeria is not supported by the SSU rDNA data. PMID- 12627868 TI - The sequence of regulatory events in the sporulation control network of Physarum polycephalum analysed by time-resolved somatic complementation of mutants. AB - The developmental decision for sporulation of Physarum polycephalum plasmodia is under sensory control by environmental factors like visible light or heat shock and endogenous signals like glucose starvation. Several hours after perceiving an inductive stimulus, plasmodia become committed to sporulation; thereby, they lose their unlimited replicative potential and execute a developmental program that involves differentiation into various cell types required to form a mature fruiting body. Plasmodia are multinuclear single cells which spontaneously fuse upon physical contact. Fusion of mutant plasmodia and cytoplasmic mixing allows complementation studies to be performed at the functional level. Mutant cells altered in their ability to sporulate in response to phytochrome activation by far-red light were cured by fusion with wild-type or other mutant plasmodia. Phytochrome activation in one plasmodium and subsequent fusion with a non-induced plasmodium revealed that complementation of the two mutations depended on (i) which of two genetically distinct plasmodial cells was stimulated; and (ii) on the delay time elapsed between stimulation and cytoplasmic mixing. Such experiments allow us to determine the kinetics and the causal sequence of the regulatory events tagged by mutation. PMID- 12627869 TI - An engineered Streptomyces hygroscopicus aph 7" gene mediates dominant resistance against hygromycin B in Chlamydomonas reinhardtii. AB - We have developed a positively selectable marker for the green alga Chlamydomonas reinhardtii using the Streptomyces hygroscopicus aminoglycoside phosphotransferase gene (aph7"). Its expression is controlled by C. reinhardtii regulatory elements, namely, the beta2-tubulin gene promoter in combination with the first intron and the 3' untranslated region of the small subunit of ribulose bisphosphate carboxylase, rbcS2. C. reinhardtii cell-wall deficient and wild-type strains were transformed at rates up to 5 x 10(-5) with two constructs, pHyg3 and pHyg4 (intron-less). Transformants selected on plates with 10 microg/ml hygromycin B exhibited diverse levels of resistance of up to 200 microg/ml that were stably maintained for at least seven months; they contained two to five copies of the construct integrated in their genomes. Transcription of the chimeric aph7" gene, correct splicing of the rbcS2 intron, and polyadenylation of the transcripts have been verified by sequencing of RT-PCR products. Average co transformation rates using pHyg3 and a second selectable plasmid were about 11%. This advocates the hygromycin-resistance plasmid, pHyg3, as a new versatile tool for the transformation of a broad range of C. reinhardtii strains without the sustained need for using auxotrophic mutants as recipients. PMID- 12627870 TI - Saville-Kent's string of pearls. PMID- 12627871 TI - Lack of association between angiotensin II type 1 receptor gene polymorphism and hypertension in Japanese. AB - Angiotensin II type 1 (AT1) receptor mediates the vasoconstriction and growth promoting effect of angiotensin II in humans. It has been reported that a polymorphism of the AT1 receptor gene (an A/C transversion at position 1166; A1166C) may be associated with essential hypertension (HT). However, several conflicting results have also been reported. Therefore, we conducted an association study between A1166C variants of the AT1 receptor gene and hypertension in the Japanese population. We genotyped this variant in 3,918 subjects (1,492 hypertensive subjects and 2,426 normotensive subjects) recruited from the Suita study. In subjects not receiving antihypertensive medication, the influence of the genotype on blood pressure values adjusted for clinical covariates was analyzed. The genotype distribution did not differ between hypertensive and normotensive subjects in either men (frequency of the C allele: 8.1% vs. 7.8%, p=0.74) or women (8.1% vs. 7.7%, p=0.60). There were no significant differences in systolic blood pressure, diastolic blood pressure, or pulse pressure among the three genotypes in either men or women who had not received hypertensive medication. Our data suggest that the A1166C polymorphism of AT1 receptor is unlikely to influence blood pressure status in the Japanese population. PMID- 12627872 TI - Relation between sodium intake and blood pressure during sleep in young men. AB - In the past, cross-sectional studies of small groups have not always shown a significant relation between habitual sodium intake and blood pressure (BP). This study examined the relation between 24-h urinary sodium excretion (Salt24, g/day) and BP during sleep in young subjects. The subjects were 132 medical students (93 males, 39 females) aged 21 to 26 years, with no renal disorder. Urine was collected for 24 h, and sodium (Na), chloride (Cl), and creatinine (Cr) excretion were measured. The 24-h urinary Cr excretion (Cr24) was compared with the Cr24 estimated from lean body mass as a guide to ensure accurate collection of 24-h urine. The natural logarithm [ln (Salt24/Cr24)] was taken as an index of sodium intake. On the same day, a portable multi-biomedical recorder (TM2425) was used to measure 24-h systolic BP (SBP)/diastolic BP (DBP). Daytime BP was calculated as the average of waking-time BP, and sleep BP was taken as statistical base BP (minimum BP) during nighttime sleep. No correlation was observed between ln (Salt24/Cr24) and daytime BP. The correlation coefficient with sleep SBP, on the other hand, was 0.34 (p<0.05) in the male group and 0.54 (p<0.01) in the male high-BP group (24-h BP>127/75 mmHg, n=28). No relation of this kind was observed in the female group. In conclusion, despite the small number of subjects, the present cross-sectional study revealed a significant positive correlation between sodium intake and sleep SBP in young men with high BP. PMID- 12627873 TI - A/C1166 gene polymorphism of the angiotensin II type 1 receptor (AT1) and ambulatory blood pressure: the Ohasama Study. AB - We previously investigated the relation between hypertension and each of three major genetic polymorphisms in the renin-angiotensin (AGT)-aldosterone system (R A-A), AGT M235T, angiotensin convert enzyme (ACE) I/D, and CYP11B2 -344C/T, by means of ambulatory blood pressure (ABP) monitoring in a general Japanese population (the Ohasama Study). A/C1166 gene polymorphism in the 3' untranslated region of the angiotensin II type 1 receptor (AT1) gene is the final remaining major target in R-A-A to be examined in the Ohasama Study population. In the present study, the AT1 A/C1166 polymorphism was genotyped by the TaqMan polymerase chain reaction (PCR) method or restriction fragment length polymorphism (RFLP) in 802 Japanese subjects aged 40 and over, who were previously genotyped for the AGT M235T, ACE D/I, CYP11B2 -344C/T polymorphisms. The AA genotype, AC genotype, and CC genotype were present in 678 (84.5%), 121 (15.1%), and 3 (0.4%) of subjects, respectively. Since the frequency of the C allele was quite low (0.079), the genotypes were classified according to the presence or absence of the C allele. Although daytime blood pressure (BP) was higher in subjects with the C allele, the difference was not statistically significant after adjusting for age, gender, body mass index, and smoking status. No significant difference was noted in the prevalence of cardiovascular diseases or nocturnal BP decline between the two groups. These results indicated that AT1 A/C1166 polymorphism was not associated with any clinical parameters associated with hypertension or atherosclerosis in the Japanese population. PMID- 12627875 TI - Cardiovascular risk factors associated with pulse pressure in a screened cohort in Okinawa, Japan. AB - We evaluated the association between pulse pressure (PP) and cardiovascular risk factors in a screened cohort. Individuals who were receiving medications for hypertension or heart disease, who had no ECG record, or who had a record of arrhythmia were excluded. In total, 8,508 subjects (5,299 men and 3,209 women; age range, 18 to 89 years) were studied. Subjects were divided into four PP classes: PP.1 (PP < or = 40 mmHg, n=2,127), PP.2 (40 < or = PP < or = 44 mmHg, n=2,127), PP.3 (44 < or = PP < or = 50 mmHg, n=2,127) and PP.4 (50 mmHg < or = PP, n=2,127). Multiple regression analysis was used for evaluating the association between PP and cardiovascular risk factor or lifestyle. In men, the regression coefficient was 0.27 for age, 2.50 for diabetes mellitus, 0.33 for uric acid, 0.20 for body mass index, 0.07 for heart rate, -0.83 for current smoking habit and 1.23 for habitual drinking. In women, the regression coefficient was 0.37 for age, 4.09 for diabetes mellitus, 0.42 for body mass index, 0.14 for heart rate, and 0.84 for habitual exercise. In both men and women, PP was significantly increased in association with an increase in the number of risk factors (diabetes mellitus, obesity, current drinking status, heart rate, and hyperuricemia). In conclusion, higher PP was associated with cardiovascular risk factors. These associations were similar in both men and women. PMID- 12627874 TI - Leptin causes nitric-oxide independent coronary artery vasodilation in humans. AB - Recent studies have shown that leptin causes vasodilation. However, it is unclear whether leptin causes coronary vasodilation in humans. To determine how leptin affects human coronary arteries and whether endothelium-derived nitric oxide (EDNO) is involved in the coronary arterial response to leptin, we infused leptin (0.3, 3 and 30 ng/kg/min) for 2 min into the left coronary ostium before and after an infusion of nitric oxide synthase inhibitor, N(G)-monomethyl-L-arginine (L-NMMA), in 11 men with angiographically normal coronary arteries. The diameter of the epicardial coronary arteries was quantitatively measured, and coronary blood flow (CBF) was calculated by quantitative angiography and Doppler flow velocity measurements. The changes in these parameters in response to leptin are expressed as the % change from the baseline values. Leptin caused coronary dilation (0.3 ng/kg/min: 2.0+/-0.5%; 3 ng/kg/min: 4.9+/-0.7%; 30 ng/kg/min: 3.8+/ 0.9%) and increased CBF (13.6+/-3.3%, 36.8+/-5.6%, and 39.2+/-7.4%, respectively). After the infusion of L-NMMA, leptin also caused coronary dilation (2.0+/-0.4%, 4.5+/-0.7%, and 4.6+/-0.8%, respectively) and increased CBF (14.6+/ 2.8%, 39.2+/-5.7%, 40.3+/-6.2%, respectively). Leptin-induced coronary vasodilation was not affected by the infusion of L-NMMA. These results suggest that leptin dilates coronary arteries in humans. Furthermore, EDNO may not contribute to leptin-induced coronary dilation. PMID- 12627876 TI - Effect of education through a periodic newsletter on persistence with antihypertensive therapy. AB - Sufficient persistence with therapy is important to achieve the desired benefits of antihypertensive therapy. This study was designed to describe the rates of persistence with antihypertensive therapy for 1 year and to assess the effect of patient education by a periodic newsletter on persistence rates in general practice in Japan. Information on 5,324 patients who received the newsletter once a month for 1 year (intervention group; 53.9% of those originally registered) and 666 patients who did not receive the newsletter (control group; 94.3% of those originally registered) was obtained from a physician questionnaire (response rate: 54.3%). The rate of persistence with antihypertensive therapy in the intervention group was similar to that in the control group (91.7% vs. 90.7%, respectively). The patient questionnaire (response rate: 28.2%) indicated that most patients consistently read the newsletter and found it useful in understanding the management of hypertension. The results of this 1-year study showed that about 90% of patients persisted with therapy for 1 year. Although most of them evaluated the newsletter favorably, no clear effect of the newsletter on their persistence with therapy was revealed. However, these results do not rule out a possible effect of education on persistence with therapy over the long-term, and thus there is need of a longer-term study employing follow-up questionnaires. PMID- 12627877 TI - Usefulness of brachial-ankle pulse wave velocity measurement: correlation with abdominal aortic calcification. AB - At present, brachial-ankle pulse wave velocity (baPWV) can be measured easily and noninvasively. We studied the correlation between aortic damage estimated by baPWV and that determined by measuring the length of abdominal aortic calcification (AAC) on X-ray films, which parameter has been significantly associated with cardiovascular morbidity and mortality. baPWV was measured using the form PWV/ankle brachial index (ABI) device in 97 patients free of end-stage renal failure or peripheral arterial disease. baPWV correlated significantly with age (r2=0.625, p<0.0001), was significantly higher in hypertensives than in normotensives (2,109+/-67 vs. 1,623+/-93 cm/s, p<0.0001), and correlated significantly with systolic blood pressure (r2=0.64, p<0.0001) and diastolic blood pressure (r2=0.397, p<0.0001). baPWV was significantly higher in diabetic patients than in nondiabetics (2,068+/-73 vs. 1,813+/-97 cm/s, p<0.05), but was similar in normolipidemic and hyperlipidemic patients. baPWV did not correlate with body mass index, fasting plasma glucose, total cholesterol, high density lipoprotein (HDL)-cholesterol, low density lipoprotein (LDL)-cholesterol or triglyceride levels, but correlated significantly with AAC length (r2=0.599, p<0.0001). Multiple regression analysis indicated that age, systolic blood pressure and AAC length were independent determinants of baPWV. Our results indicate that baPWV is useful for estimating aortic damage and could be a potentially useful predictor of vascular morbidity and mortality. PMID- 12627878 TI - Tissue-specific impairment of insulin signaling in vasculature and skeletal muscle of fructose-fed rats. AB - The relation between insulin resistance/hyperinsulinemia and cardiovascular diseases has attracted much attention. Insulin affects not only glucose metabolism, but also protein synthesis and cell growth. Insulin stimulates both the phosphatidylinositol 3-kinase (PI3-K) and mitogen-activated protein kinase (MAPK) pathways, but the relationship between cardiovascular disease and selective insulin signal pathways is unclear. We investigated the tissue specificity and intracellular signal transduction selectivity of insulin resistance in the vasculature and skeletal muscle of fructose-fed rats (FFR). Sprague-Dawley rats were fed either normal rat chow (control rats) or fructose rich chow. Normal saline with or without 1,000 (microg/kg) insulin was injected, and then the thoracic aorta or soleus muscle was removed under anesthetization. Insulin-induced tyrosine phosphorylation of insulin receptor beta subunit (IRbeta) and insulin receptor substrate-1 (IRS-1) and tyrosine/threonine phosphorylation of p44/42 MAPK (ERK-1/2) were evaluated. There were no significant differences in the degree of phosphorylation of IRbeta or ERK-1/2 in the thoracic aorta or in the soleus muscle between FFR and controls. However, tyrosine phosphorylation of IRS-1 in the soleus muscle of FFR was significantly reduced to 80% (p<0.001) of that in controls. The results suggest that PI3-K pathway in skeletal muscle is selectively impaired in FFR, and this impairment may induce hyperinsulinemia, which in turn may stimulate the MAPK pathway and lead to atherosclerosis. Thus PI3-K pathway may be one of the factors underlying the onset of cardiovascular disease in patients with insulin resistance. PMID- 12627879 TI - Lipoxygenase products regulate nitric oxide and inducible nitric oxide synthase production in interleukin-1beta stimulated vascular smooth muscle cells. AB - In cultured vascular smooth muscle cells (VSMCs), interleukin-1beta (IL-1beta) stimulates inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) production. IL-1beta also activates phospholipase A2 (PLA2), and induces lipoxygenase (LOX) and cyclooxygenase-2 (COX-2). The present study investigated whether these metabolites are involved in the regulation of IL-1beta-induced NO production and iNOS expression. Pretreatment with ONO-RS-082, the secretory PLA2 (sPLA2) inhibitor, at 1 to 10 micromol/l reduced IL-1beta-stimulated nitrite production and iNOS expression. Nordihydroguaiaretic acid (NDGA, 1 to 10 micromol/l), the LOX inhibitor, also reduced IL-1beta (10 ng/ml)-stimulated nitrite production and iNOS expression in a dose-dependent manner. Exogenous 12(S)-hydroxyeicosatetraenoic acids (HETE) enhanced the IL-1beta-stimulated nitrite production and iNOS expression. On the other hand, the COX inhibitors, indomethacin and NS-398, had little effect on nitrite production or iNOS expression. These results suggest that LOX products play important roles in the regulation of stimulus-induced NO production in VSMCs. PMID- 12627881 TI - Disconnecting surgical treatment of hypothalamic hamartoma in children and adults with refractory epilepsy and proposal of a new classification. AB - A series of 17 patients aged from 9 months to 32 years with refractory epilepsy due to hypothalamic hamartoma were treated by total removal (one case) and disconnection (16 cases) between 1997 and 2002. The mean age at seizure onset was 16 months. Sixteen patients had gelastic seizures, 14 had partial seizures and three had generalized tonic-clonic seizures. The mean seizure frequency was 21 per day. Four patients had borderline intelligence quotient and the others were mentally retarded. Five patients presented with precocious puberty, one with acromegaly, and four suffered from obesity. Brain magnetic resonance imaging, performed at least twice in each patient, showed the hamartoma as a stable homogeneous interpeduncular mass implanted either on the mammilary tubercle or on the wall of the third ventricle with variable extension to the bottom. Ictal single photon emission computed tomography, performed in four patients, showed hyperperfusion within the hamartoma in two patients. Twenty-five operations were performed in the 17 patients. The first patient underwent total removal of the hamartoma, whereas the following 16 patients underwent disconnection through open surgery (14 procedures) and/or endoscopy (9 procedures). Eight patients became seizure-free, one patient had only brief gelastic seizures, and eight patients were dramatically improved with a mean follow up of 18.6 months (8 days to 43 months). Surgery was safe in all but two patients: the first patient had transient hemiplegia and the third cranial nerve paresis, and the other developed hemiplegia due to ischemia of the middle cerebral artery territory. The quality of life, and behavior and school performance were greatly improved in most patients. Our series illustrates the feasibility and relative safety of disconnection surgery for hypothalamic hamartomas with seizure relief in 53% of patients and dramatic improvement in the others. Surgical observations led us to propose a new anatomical classification according to the anatomical relationship between the hamartoma and the adjacent hypothalamus and third ventricle. Endoscopic disconnection seems to be a very safe way to treat hamartomas in intraventricular locations. PMID- 12627880 TI - Effects of the anti-platelet aggregation drug dilazep on cognitive function in Dahl salt-sensitive rats. AB - Among the consequences of the increasing prolongation of lifespan is a worldwide increase in the number of cases of dementia or impaired cognition. In the present study, to test the hypothesis that mechanisms independent of high blood pressure are involved in maintaining cognitive function, we assessed the effects of long term dilazep treatment on cognitive dysfunction in normotensive Dahl salt sensitive (Dahl S) rats fed a low-salt diet, using the standard passive avoidance test. Normotensive Dahl S rats fed a 0.3% NaCl diet were treated for 6 months with low-dose dilazep (2.5 microg/ml in drinking water) or high-dose dilazep (12.5 microg/ml). Systolic blood pressure was within normotensive range throughout the study and did not differ among the experimental groups. The results of the passive avoidance test revealed that dilazep treatment attenuated the decline of latency time relative to that in the untreated control rats (control latency time, 235 s; low-dilazep group, 389 s; high-dilazep group, 397 s), suggesting that the cognitive function of normotensive Dahl S rats was improved by dilazep treatment. This improvement of cognition was associated with significant increases in the number of neuronal cells in the hippocampal region and with an increase in capillary length in dilazep-treated Dahl rats. In addition, the dilazep treatments significantly attenuated arteriolar injury of glomeruli in the kidney. These data suggest that dilazep treatment, through vascular and non-vascular effects, maintains the brain function in Dahl S rats susceptible to vascular injury and organ dysfunction. PMID- 12627882 TI - Artificial cerebral aneurysm model for medical testing, training, and research. AB - Artificial models of cerebral aneurysms for medical training and testing of medical devices were constructed from corrosion casts of the main cerebral arteries of a human specimen. Three aneurysms with a variety of shapes were simulated at typical locations. Rigid and soft models were made of silicone using the "lost wax" technique. The transparent silicone models were anatomically accurate and reproducible copies of human vascular casts. These models could be connected in a closed circuit that used an electric pump to simulate pulsatile flow. Endovascular procedures and surgical clip application were performed under fluoroscopic or direct visual control. Surgical clipping, endoluminal coil manipulation, and consecutive hemodynamic changes were visualized by digital subtraction angiography and direct observation. The model provides trainee surgeons with an understanding of clinical conditions. New medical devices, such as platinum coils, would be experimentally implanted in the model under stable conditions. These anatomically accurate and reproducible models of cerebral vasculature and aneurysms are valuable for medical testing, training, and research. PMID- 12627883 TI - Subarachnoid hemorrhage induces Na+/myo-inositol cotransporter in the rat brain. AB - Neurons and glial cells respond to extracellular hyperosmolarity by accumulating small organic solutes, called "osmolytes." Na+/myo-inositol is one of the major organic osmolytes in the brain and Na+/myo-inositol cotransporter (SMIT) regulates extracellular Na+/myo-inositol content. Subarachnoid hemorrhage (SAH) is an osmotic stress-inducing event of the brain. The expression of SMIT messenger ribonucleic acid (mRNA) and protein was investigated with in situ hybridization and immunohistochemistry in rat brains with SAH induced by endovascular perforation. SMIT riboprobe was raised from a 490-bp rat SMIT complementary deoxyribonucleic acid. Anti-SMIT antibody was raised in rabbits. SMIT mRNA was expressed strongly in the cortex, hippocampus, and hypothalamus of the perforated side at 6 to 24 hours after SAH. Mild upregulation was noted in the contralateral cortex, hippocampus, and hypothalamus. The ventral aspect of the pons showed mild upregulation. Microautoradiography and immunostaining showed SMIT expression mainly in the neurons, but also in some non-neural cells in the hippocampus. The present results indicate that diffuse osmotic stress occurs in the host brain after SAH. PMID- 12627884 TI - Unusual accessory cranial sutures in pediatric head trauma--case report. AB - A one-year, 9-month-old boy presented with pediatric head trauma associated with unusual accessory cranial sutures. Radiography demonstrated unusual bilateral longitudinal linear bone defects extending from the foramen magnum to the mendosal sutures, and bilateral transverse linear bone defects around the foramen magnum. No swelling or soreness were found in the occipital area, and there was no past history of head trauma. Therefore, the bone defects were considered to be accessory cranial sutures. Complex developmental patterns of the occipital bone and the considerable normal variation of sutures may simulate fractures around the foramen magnum. The present case of accessory cranial sutures is another example. PMID- 12627886 TI - Surgical treatment of spontaneous intracerebral hemorrhage in a full-term infant with coagulopathy--case report. AB - An 11-week-old male infant presented with intracerebral hemorrhage associated with coagulopathy manifesting as left hemiparesis, lethargy, and vomiting. Computed tomography demonstrated extensive right frontoparietal intracerebral hemorrhage extending into the ventricular system. Liver function tests revealed abnormal values of transaminases and bilirubin. Blood coagulation studies showed prolonged prothrombin time (PT) and activated partial thromboplastin time (APPT). PT and APTT immediately normalized after the administration of vitamin K and fresh frozen plasma. Right parietal craniotomy and evacuation of the hematoma were performed because of the deterioration in consciousness and left hemiparesis. No vascular abnormality was observed in the hematoma cavity. After surgery, he became alert and the left hemiparesis improved. There is a risk of intracerebral hemorrhage due to vitamin K deficiency even if prophylactic administration of vitamin K was given. Surgical treatment should be considered for the treatment of infantile spontaneous intracerebral hemorrhage, especially if neurological deterioration is present. PMID- 12627885 TI - Subcortical hemorrhage caused by methamphetamine abuse: efficacy of the triage system in the differential diagnosis--case report. AB - A 32-year-old woman was brought to the emergency room with hemiplegia on the left and consciousness disturbance. Her prior medical history and the circumstances of the onset were unknown. Brain computed tomography showed intracerebral hemorrhage (ICH) with a midline shift of more than 10 mm in the right parietal lobe. Cerebral angiography failed to show any vascular anomalies. Urine analysis with the triage system, a qualitative screening test for psychotropic drug abuse, showed positive reaction for amphetamines. Subsequent laboratory examination confirmed a highly elevated serum concentration of methamphetamine. The patient underwent evacuation of the hemorrhage via a craniotomy, and was discharged 40 days after admission. Abuse of illegal drugs including amphetamines among young adults is increasing in many developed countries, and the suspicion of possible drug abuse should always be raised in young patients with angiographically negative ICH. A urinalysis screening test for psychotropic agents should be a part of routine emergency room diagnostic procedures for such patients. PMID- 12627887 TI - Osteosarcoma arising from the skull--case report. AB - A 20-year-old male presented with an osteosarcoma in the right parieto-occipital bone occurring as a painless occipital lump which had rapidly enlarged in the 6 months prior to admission. The neuroimaging appearance resembled intraosseous meningioma. Gross total resection of the tumor was achieved. The final histological diagnosis was osteosarcoma. Osteosarcomas of craniofacial region have a better prognosis than those of the skeletal bones, and distant metastasis is rare. Local recurrence is the most significant factor contributing to poor outcome. Complete excision with negative margins is the key to a better outcome. Adjuvant therapy may be an option in cases of incomplete excision. Advances in target chemotherapy may diminish the significant morbidity associated with these lesions. PMID- 12627888 TI - Cerebellar hemangioblastoma associated with primary hyperparathyroidism--case report. AB - A 57-year-old man presented with a posterior fossa hemangioblastoma associated with primary hyperparathyroidism. The hemangioblastoma was completely removed. Further imaging found parathyroid and thyroid tumors which were resected. Hyperparathyroidism might be a manifestation of von Hippel-Lindau (VHL) disease, but sequencing of the VHL gene of the constitutional deoxyribonucleic acid of the hemangioblastoma was negative. The present association is rare, but the hemangioblastoma and the parathyroid adenoma may have some germ-line mutation in common. PMID- 12627889 TI - Schwannoma in the crural cistern removed without permanent functional deficits- case report. AB - A 33-year-old male presented with a rare intracranial oculomotor schwannoma not associated with neurofibromatosis manifesting as an 8-month history of right oculomotor nerve paresis. Neuroimaging examinations revealed a 4-cm mass in the right crural cistern. The tumor was totally removed. The right oculomotor nerve paresis resolved 7 months after surgery. Removal of the oculomotor or trochlear schwannoma usually results in parent nerve paresis, but subcapsular tumor resection in our case probably allowed complete recovery of the nerve function. PMID- 12627890 TI - Pituitary abscess showing high uptake of thallium-201 on single photon emission computed tomography--case report. AB - A 32-year-old female presented with a rare case of pituitary abscess manifesting as homonymous hemianopsia. Serum prolactin level was slightly high (40.8 ng/ml). Magnetic resonance (MR) imaging showed the content of the lesion as homogeneously isointense on the T1-weighted images and hyperintense on the T2-weighted images. The capsule of the lesion, which appeared thin and smooth, was enhanced by gadolinium. Dural enhancement around the sella turcica was also recognized. Thallium-201 single photon emission computed tomography (201Tl SPECT) showed homogeneous high accumulation in the pituitary region on both the early and delayed images. The lesion was treated via a transnasaltranssphenoidal approach. The cystic lesion contained pus and the capsule consisted of normal pituitary gland with inflammatory changes. The patient was treated with antibiotics for 3 weeks and the pituitary abscess was cured completely. Pituitary abscess can be differentiated from pituitary adenoma as lesion with a homogeneous high uptake on 201Tl SPECT on both the early and delayed images, and no enhancement of the central portion on MR images. PMID- 12627891 TI - The biomass dynamics of cyanobacteria in an annual cycle determined by confocal laser scanning microscopy. AB - We recently published a method for estimating cyanobacteria biomass in delta microbial mats from the Ebro river using confocal laser scanning microscopy (CLSM). The present paper uses this method for identifying different groups of cyanobacteria and for determining their biomass dynamics. Microcoleus chthonoplastes and the Lyngbya-Oscillatoria were the most important contributors to the cyanobacterial biomass throughout the study. Biomass values ranged from 1.29 to 6.55 mgC/cm2 sediment for Microcoleus chthonoplastes, and from 128 microgC/cm2 to 3.16 mgC/cm2 sediment for Lyngbya-Oscillatoria. This technique is useful for determining biomass and for studying filamentous cyanobacteria as well as ramified eukaryotic cells. Confocal serial sections through the samples can be obtained. Two-dimensional images from the samples can be used to calculate the biomass of individual cells. PMID- 12627892 TI - Evaluation of fan-pattern spray nozzle wear using scanning electron microscopy. AB - Worn nozzles on spray equipment severely affect the efficiency of crop management systems while causing unnecessary pesticide contamination of non target areas. Scanning electron microscopy (SEM) that has been applied for direct measurement of pesticide deposition, was used to observe worn and unused brass and stainless steel fan-pattern spray nozzles. Wear and other changes were observed in both nozzle materials. Scanning electron microscopy can provide nozzle manufacturers with greater insight and needed information on nozzle mechanics to improve performance. More reliable delivery of pesticide spray should enhance integrated pest and disease management and crop protection for growers. PMID- 12627893 TI - Microstructure of Monoplacophora (Mollusca) shell examined by low-voltage field emission scanning electron and atomic force microscopy. AB - The shell of Micropilina arntzi (Mollusca: Monoplacophora), a primitive molluscan class, was examined by using field emission scanning electron microscopy (FESEM) at low voltage and atomic force microscopy (AFM). The use of these two techniques allowed the observation of fine details of Micropilina arntzi shell and contributed to bring new features concerning the study of molluscan shell microtexture. Imaging with low-voltage FESEM provided well-defined edge contours of shell structures, while analyzing the sample with AFM gave information about the step height of stacked internal structures as well as the dimension of the particles present in their surface at a nanometric level. The shell microstructure of Monoplacophora species presents different patterns and may be a taxonomic implication in the systematic studies of the group. PMID- 12627894 TI - Exploring the structure of a hydrogen cyanide polymer by electron spin resonance and scanning force microscopy. AB - Aqueous solutions of potassium cyanide and ammonium hydroxide are known to yield a heterogeneous cyanide polymer, containing paramagnetic sites and biologically significant substructures including polypeptides. Here, such solutions were used to prepare various samples of polymer for study by X-band and W-band electron spin resonance (ESR), scanning electron microscopy (SEM), and scanning force microscopy (SFM). Elemental composition of a typical sample of the polymer was C 35.2%, N-38.47%, 0-14.51%, and H-4.13%, exposing the polymer to 6M HCl hydrolyzed portions of the polymer and released glycine and traces of other amino acids. The X-band ESR spectra consist of a single slightly asymmetric line centered at g = 2.003; spin concentration measurements made at X-band using a nitroxide radical standard yield approximate radical concentrations of 10(18) spins/gm. W-band ESR indicates the presence of a single rhombic paramagnetic site with g(x) = 2.0025, g(y) = 2.0030, and g(z) = 2.0048 and the possibility of small 14N hyperfine splittings. The ESR spin echo studies yield a longitudinal relaxation time, Tl of 75 microS and a short-phase memory relaxation time, Tm, of about 300 nS. Scanning electron microscopy studies of the polymer show that it is made of ellipsoidal particles about one micron in size. The particles tend to clump together when suspended in aqueous solution. The particles disperse and dissolve in dimethyl sulfoxide (DMSO); when these solutions dry on microscope slides, optical microscopy shows a branched island morphology for the polymer. This morphology is reminiscent of snowflakes and is identified as dendritic. Phase contrast SFM of the dendritic arms show a striking segregation and ordering of various components of the polymer. Paramagnetic sites are conserved in the series of steps leading to dendritic structures. PMID- 12627895 TI - Correction for piezoelectric creep in scanning probe microscopy images using polynomial mapping. AB - We describe a method for using polynomial mapping to correct scanning probe microscope images for distortion due to piezoelectric creep. Because such distortion varies from image to image, this method can be used when the actual locations of some features within an image are known absolutely, or in a series of images in which the actual locations of some features are known not to vary. While the general case of polynomial mapping of degree N requires the determination of 2(N+ 1)2 matrix elements by regression, we find that by understanding the mechanism by which piezoelectric creep distorts scanning probe microscope images, we can fix most of these coefficients at 0 or 1 a priori, leaving only 2(N+ 1) coefficients to be determined by regression. We describe our implementation of this strategy using the Interactive Data Language (IDL) programming language, and demonstrate our technique on a series of atomic force microscopy (AFM) images of diblock copolymer microdomains. Using our simplified scheme, we are able to reduce the effects of distortion in an AFM image from 5% of the scan width to a single pixel, using only five reference points. PMID- 12627896 TI - Scanning electron microscopy analysis of corrosion degradation on tinplate substrates. AB - The degradation of electrolytic tinplate used in food containers was analysed and evaluated, using scanning electron microscopy and electrochemical measurements of microcorrosion and ion dissolution by atomic absorption to prevent food contamination caused by metal traces and to increase the durability of such tinplates. PMID- 12627897 TI - Computer-assisted stereology: point fraction of lung parenchyma and alveolar surface density in fetal and newborn sheep. AB - The aim of this study was to determine the accuracy of a new computer-assisted stereological technique in obtaining structural information of the lung. We compared the point fraction of lung parenchyma (Pp) and alveolar surface density (Sv) obtained by established manual point/intercept counting methods and compared them with those obtained using a computer-assisted method. Lung tissues obtained from normally grown fetal sheep (n = 6) and from newborn lambs with severe lung hypoplasia (n = 5) were inflation fixed via the trachea and processed for light microscopy. In verification-of-technique experiments, Pp and Sv correlated well with known values. There was a significant linear correlation between manual and computer-assisted stereological measurements for values of Pp (r2 = 0.92) and Sv (r2 = 0.98). Our data lead us to believe that the computer-assisted stereological technique described in this study provides accurate estimates of Pp and Sv and hence may be a valuable tool for evaluating the effects of factors upon structural development of the lung. PMID- 12627898 TI - Combined atomic force microscopy and scanning tunneling microscopy imaging of cross-sectioned GaN light-emitting diodes. AB - Cross-sectional scanning tunneling microscopy (STM) was combined with atomic force microscopy (AFM) over the same area to characterize a cross-sectioned GaN light emitting diode. Because GaN is typically grown on a non-native substrate and also forms a wurtzite crystal structure, a cryogenic cleaving technique was developed to generate smooth surfaces. The depletion region surrounding the p-n junction was clearly identified using STM. Furthermore, by imaging under multiple sample biases, distinctions between the n-doped and p-doped GaN could be made. PMID- 12627899 TI - Safe motherhood in Pakistan: past failures, future challenges. PMID- 12627900 TI - Maternal mortality in Pakistan--compilation of available data. AB - OBJECTIVE: To compile available information - both hospital and community based- on the maternal mortality ratios (MMRs) and the causes of maternal deaths in Pakistan METHODS: Data was collected from articles published in Medical Journals, in Proceedings of Conferences and of Workshops as well as from Newsletters of the National Committee for Maternal Health (NCMH). The data was then tabulated in order of the year of publication. RESULTS: The MMRs reported from hospitals vary between 17 in a private tertiary hospital to 2,736 in a government tertiary hospital. In the community the range is from 160 in Sindh to 673 in Khuzdar. Data for different periods of time from three tertiary public hospitals, two in the south and one in the north of Pakistan, show no decrease in MMRs. The trend remains the same. In the hospitals over 80% of the deaths are due to direct causes. The most common cause of death is hemorrhage, followed by eclampsia and sepsis. The causes of death in the community are also the same though the proportions vary. Direct causes account for 78.1% of deaths, hemorrhage being the most common followed by sepsis, eclampsia, rupture of the uterus, and abortions. CONCLUSIONS: Available data shows that the Maternal Mortality Ratios in Pakistan continue to remain high despite attention paid to maternal health. To address the unacceptable state of maternal health, a policy change with a more comprehensive and integrated approach to maternal health services is needed. Strategies need to be developed which are short term, medium term and long term. This includes competency based training, deployment and supervision of a cadre of skilled attendants for delivery at the community level. In addition upgrading of health care systems especially availability of emergency obstetric care is urgently required. PMID- 12627901 TI - Determination of alpha-1 antitrypsin genetic deficiency in duodenal ulcer by polymerase chain reaction. AB - OBJECTIVE: To confirm alpha-1-AT deficiency status in duodenal ulcer using a combination of PCR and restricted enzyme digestion. METHODS: Fifty patients with endoscopically proven duodenal ulcer and hundred controls with no signs of the disease were included. Alpha-1-AT phenotypes were confirmed by polymerase chain reaction followed by restriction enzyme digestion. RESULTS: Alpha-1-AT concentration in duodenal ulcer patients showed a mean value of 2.12 +/- 0.11 g/l (range: 0.52-3.95 g/l, p < 0.05). In controls this was 2.47 +/- 0.08 g/l (range: 0.52-5.0 g/l). Among the controls 70% had the MM phenotype, 28% M1 M2 and 2% FM. In duodenal ulcer, MM predominates (70%), followed by M1 M2 (18%), SS (4%), SZ (4%), ZZ (2%) and MZ (2%). CONCLUSION: Alpha-1 AT deficiency was found in 10% of duodenal ulcer patients. DNA analysis more accurately resolved the phenotypes as S and Z mutations. PMID- 12627902 TI - Serum and CSF immunoglobulins G, A and M in 37 intracranial tumors. AB - OBJECTIVE: To estimate different proteins and immunoglobulins as humoral immune response in patients with intracranial tumours. METHODS: Cellulose acetate membrane electrophoresis for estimation of different proteins and radialimmunodiffusion (RID) for measurement of serum and CSF immunoglobulins were used. RESULTS: Thirty seven patients with different types of tumours showed increase in serum alpha II globulin, significant decrease of IgG, in patients when compared with controls, significant decrease of IgG in malignant tumours compared with benign tumours. CSF albumin, gamma globulin, immunoglobulins G,A,and M were increased. CONCLUSION: Alpha II globulin, steroid therapy and active neoplasia may be responsible for decreased serum immunoglobulins, while raised gamma globulin and immunoglobulins with intact BBB indicates capability of CNS to produce immunoglobulins. PMID- 12627903 TI - Birth asphyxia--analysis of 235 cases. AB - OBJECTIVE: To look for risk factors leading to birth asphyxia in new borns admitted in a tertiary care unit STUDY DESIGN: Retrospective analysis. SETTING: Neonatal Unit of National Institute of Child Health, Karachi from 1st January, 2001 to 31st August, 2001. PATIENTS AND METHODS: Records of 235 new borns admitted with birth asphyxia during this period were analyzed. Variables studied included antenatal care, period of gestation, place of delivery, mode of delivery, birth weight and age at arrival in the hospital. RESULTS: Majority (71%) of mothers were booked and had antenatal care, similarly most (88%) of the babies were born at term and 75.3% were delivered in maternity homes or hospitals. Caesarian sections were performed in 14% cases and rest were all vaginal deliveries. Fifty seven percent newborns weighed more than 2.5 kg and 22.5% presented to the hospital within 6 hours of birth. CONCLUSION: Birth asphyxia occurring in such a high number of booked cases delivered at term with good weight, reflects the poor perinatal services offered in those maternity homes or hospitals. It is recommended that trained personnel and neonatal resuscitation equipment should be made mandatory in all maternity homes/hospitals. PMID- 12627904 TI - Suprapubic cystostomy: urinary tract infection and other short term complications. AB - AIMS: To evaluate the frequency of urinary tract infection in patients with suprapubic cystostomy and other complications of the procedure within 30 days of placement. METHODS: Patients characteristics, indication and types of cystostomy and short term (within 30 days); complications were analyzed in 91 patients. Urine analysis and culture was done in all patients to exclude those with urinary tract infection. After 15 and 30 days of the procedure, urine analysis and culture was repeated to evaluate the frequency of urinary tract infection. The prevalence of symptomatic bacteriuria with its organisms was assessed. Antibiotics were given to the postoperative and symptomatic patients and the relationship of antibiotics on the prevention of urinary tract infection was determined. RESULTS: Of the 91 cases 88 were males and 3 females. The mean age was 40.52 +/- 18.95 with a range of 15 to 82 years. Obstructive uropathy of lower urinary tract.was present in 81% cases and 17 (18.6%) had history of trauma to urethra. All these cases had per-urethral bleeding on examination while x-ray urethrogram showed grade II or grade III injury of urethra. Eighty two of the procedures were performed per-cutaneously and 7 were converted to open cystostomies due to failure of per-cutaneous approach. Nine patients had exploratory laparotomy. Duration of catheterization was the leading risk factor for urinary tract infection found in 24.1% at 15 days and 97.8% at 30 days. There was low prevalence of symptomatic urinary tract infection. E.Coli was the prevalent organism. Antibiotics did not play a significant role in controlling catheter associated urinary tract infection. Blockage and macroscopic hematuria (self limiting) were the predominant complications encountered. CONCLUSION: Duration of catheterization is the leading risk factor for the development of urinary tract infections. Catheter-associated bacteriuria are usually asymptomatic. Macroscopic hematuria and blockage of catheter are frequent which can be managed promptly without surgical intervention. PMID- 12627905 TI - Cancer patterns in Quetta (1998-1999). AB - INTRODUCTION: Quetta, the capital of Baluchistan, is located at latitude 30.25; longitude 67.00. It has a population of 759,245; 425,474 males (56%) and 333,771 females (44%); Census 1998. The majority of residents are Persian or Baluchi speaking Baluchs. METHODOLOGY: The cancer cases from Quetta reported to the Karachi Cancer Registry were reviewed. The data included cases from the Aga Khan University Hospital (AKUH), Pathology Department (Quetta collection point) and health care facilities in Karachi. The residents of Baluchistan were ascertained and cancer cases residents of Quetta identified with the help of recorded addresses and retraceable telephone numbers. RESULTS: During a 2-year period, 1st January 1998 to 31st December 1999 the Karachi Cancer Registry received 1077 cancer notifications from Quetta. Approximately half the cases were registered from the AKUH Quetta collection point. Others were the referral patterns in Karachi. The age-standardized incidence rate (ASR) of cancer, all sites (1998-99) was 137.0 for males and 92.8/100,000 for females. The commonest cancer in the males and females was cancer of the esophagus (ICD-10 categories C15; males- ASR 25.5, 17.2%; females-ASR 23.4,23.1%). One ofthe highest in the world, for both genders. Incidence of cancer breast in the females (ICD-10 categories C50; ASR 11.8, 13.3%) was low. CONCLUSION: The cancer data from Quetta is quite distinct from the cancer pattern of Karachi. It represents only a part of the cancer burden of Quetta; nonetheless it gives a glimpse into the cancer profile of Baluchistan. The high incidence of esophageal cancer indicates an extension of the geographical limits of the esophageal cancer belt. PMID- 12627907 TI - Diabetes mellitus: the role of the laboratory--an update. PMID- 12627906 TI - Management of atrial fibrillation in older patients. AB - INTRODUCTION: There has been a rapid growth in the elderly population and atrial fibrillation (AF) is the most common sustained cardiac arrhythmia among older patients. There is considerable diversity in the management of AF and different treatment regimens have been proposed. These treatments are influenced by multiple factors. However there is no information available on the management of AF in very elderly patients. Most evidence is based upon research in younger people which cannot be extrapolated to older patients. METHOD: The study was a prospective assessment of patients over 77 years admitted to a district general hospital. New onset AF was defined as AF of less than 3 months duration and chronic AF was defined as AF for more than 3 months. The electrocardiograms of all patients were reviewed following admission and patients were included provided they had AF on admission or during their hospital stay. RESULTS: Over the six months 783 patients were admitted and of those 153 patients had AF (20%). The mean age was 85.2 years and in 33% of patients AF was the primary cause for admission. New onset AF was noted in 35% of patients and 65% had chronic AF. Intermittent AF was noted in 10% and no patient met the criteria for lone AF. Most patients presented with multiple medical problems averaging nearly five pathologies per patient. Though 41% of patient received thromboprophylaxis, most had inappropriate dosage thus highlighting the problems surrounding less rigid patient selection or inadequate monitoring. Twentysix percent of patients died during their hospital stay but no deaths were recorded in patients who reverted to sinus rhythm. CONCLUSION: Elderly patients with AF are a diverse group with limited treatment options due to delayed hospital admission and multiple pathology. Chemical cardioversion is safe in the short term and those who cardiovert have better prognosis. Awareness is required for adequate dosage and monitoring of antithrombotic medications. PMID- 12627908 TI - Post-chemotherapy residual mass in stage IIC seminomatous testicular tumor. PMID- 12627909 TI - Symptomatic granulomatous prostatitis following bacillus Calmette-Guerin immunotherapy for bladder cancer. PMID- 12627910 TI - Mature cystic teratoma presenting as the largest mediastinal tumour ever recorded in the history of medicine. PMID- 12627912 TI - Chondroid choristoma of tongue--a rare entity. PMID- 12627911 TI - View box case--12. Osteomyelitis. PMID- 12627913 TI - Disaster medicine: status, roles, responsibilities, and needs. PMID- 12627914 TI - Instrument development and evaluation of domestic preparedness training for first responders. AB - INTRODUCTION: In the wake of domestic terrorists attacks on 11 September 2001 and subsequent bioterrorist events employing anthrax, there no longer can be any debate about the potential for attacks employing Nuclear, Biological, or Chemical (NBC)/Weapons of Mass Destruction (WMD). As one way of acknowledging this long standing threat and, in a concerted effort to mitigate the effects of possible future domestic NBC/WMD terrorist attacks, the US Department of Defense (DOD) and other US governmental agencies already had mounted an effort to provide Domestic Preparedness Training for First Responders in urban centers throughout the USA. METHODS: A paper and pencil questionnaire specifically designed to evaluate the effectiveness of Domestic Preparedness Training for Emergency First Responders has been developed. An earlier version of this instrument was piloted with a convenience sample of firefighters and paramedics (n = 78) in a northwest state. Based on replies to the pilot questionnaire, a pool of 27 items based on the objectives and content of the NBC/WMD Domestic Preparedness Awareness and Operations courses (plus additional background and appraised competency items) were selected for inclusion in a Domestic Preparedness Questionnaire (DPQ). RESULTS: This paper first describes the essential psychometric properties of the DPQ based on replies from baseline and follow-up samples (n = 206 and n = 246 respectively) of urban firefighters and paramedics employed by a metropolitan city in a northwest state. The DPQ was employed to evaluate the outcomes of Domestic Preparedness training provided to a sample of urban fire-service personnel. The DPQ documented significant improvements in a group of "DP trained" urban firefighters (n = 80) both in their awareness and operations content knowledge as well as in their perceived competencies to respond to acts of biological, chemical, or nuclear terrorism "in their own community" at four months post-training. A comparison group of "Not DP-trained" firefighters (n = 78) showed no statistically significant changes on these DPQ indices, suggesting that the documented improvements in the "DP-trained" firefighters on the DPQ were not due to "test reactivity" or to "historical" factors. CONCLUSION: The findings suggest that the DPQ has adequate inter-item and test-retest reliability, possesses concurrent validity, and appears to be a sensitive measure of the Domestic Preparedness Training provided for urban firefighter and paramedic First Responders. PMID- 12627915 TI - Post-flood--infectious diseases in Mozambique. AB - INTRODUCTION: The types of medical care required during a disaster are determined by variables such as the cycle and nature of the disaster. Following a flood, there exists the potential for transmission of water-borne diseases and for increased levels of endemic illnesses such as vector-borne diseases. Therefore, consideration of the situation of infectious diseases must be addressed when providing relief. The Japan Disaster Relief (JDR) Medical Team was sent to Mozambique where a flood disaster occurred during January to March 2000. The team operated in the Hokwe area of the State of Gaza, in the mid-south of Mozambique where damage was the greatest. METHODS: An epidemiological study was conducted. Information was collected from medical records by abstracting data at local medical facilities, interviewing in habitants and evacuees, and conducting analyses of water. RESULTS: A total of 2,611 patients received medical care during the nine days. Infectious diseases were detected in 85% of all of patients, predominantly malaria, respiratory infectious diseases, and diarrhea. There was no outbreak of cholera or dysentery. Self-reports of the level of health decreased among the flood victims after the event. The incidence of malaria increased by four to five times over non-disaster periods, and the quality of drinking water deteriorated after the event. CONCLUSIONS: Both the number of patients and the incidence of endemic infectious diseases, such as malaria and diarrhea, increased following the flood. Also, there was a heightening of risk factors for infectious diseases such as an increase in population, deterioration of physical strength due to the shortage of food and the temporary living conditions for safety purposes, and turbid degeneration of drinking water. These findings support the hypotheses that there exists the potential for the increased transmission of water borne diseases and that there occurs increased levels of endemic illnesses during the post-flood period. PMID- 12627916 TI - First-aid training and bystander actions at traffic crashes--a population study. AB - INTRODUCTION: Traffic crashes constitute a major, worldwide public-health problem that cause disabilities, life-long suffering, and huge economic losses. When a person is injured in a traffic crash, actions taken by bystanders often are of crucial importance. To perform first-aid actions in a correct manner, bystanders, often laypersons, need both the courage and the knowledge to do so. For preventive purposes, society spends large resources to inform and educate the public in order to enhance people's ability to take correct actions. However, there only is little information on the rate in a population of persons who have had first-aid training, have been bystanders at a traffic crash, on the actions taken by such persons, and on effects of first-aid training on patient care. OBJECTIVE: The aim of this study was to acquire knowledge about: (1) the prevalence of first-aid training; (2) the incidence of being a bystander and of the first aid provided at traffic crashes and other emergencies; and (3) the impact of first-aid training on the risks people take in road traffic. METHODS: A questionnaire was administered to 2,800 randomly selected persons aged 18-74 years. RESULTS: The response rate was 67.5%. During the previous five years, 39% of the population had received first-aid training, with a higher rate among younger individuals and those with a higher education. After training, 30% of the respondents had used their skills, and 41% took fewer risks in traffic, particularly those who were older or had a lower level of education. Fourteen percent of those with training (significantly more men) had been bystanders at a traffic crash. At 20% of the crashes, a bystander had administered first aid, and one-third of those who provided such assistance had had use of their training. CONCLUSION: Intensified first-aid training of the general public could lead to citizens who are more cautious in traffic and to bystanders who provide more immediate and adequate first aid at traffic crashes and other emergencies. PMID- 12627917 TI - Effectiveness of a Glasgow Coma Scale instructional video for EMS providers. AB - INTRODUCTION: The Glasgow Coma Scale (GCS) is the standard measure used to quantify the level of consciousness of patients who have sustained head injuries. Rapid and accurate GCS scoring is essential. OBJECTIVE: To evaluate the effectiveness of a GCS teaching video shown to prehospital emergency medical services (EMS) providers. METHODS: Participants and setting--United States, Mid Atlantic region EMS providers. Intervention--Each participant scored all of the three components of the GCS for each of four scenarios provided before and after viewing a video-tape recording containing four scenarios. Design--Before-and after single (Phase I) and parallel Cohort (Phase II). Analysis--Proportions of correct scores were compared using chi-square, and relative risk was calculated to measure the strength of the association. RESULTS: 75 participants were included in Phase I. In Phase II, 46 participants participated in a parallel cohort design: 20 used GCS reference cards and 26 did not use the cards. Before observing the instructional video, only 14.7% score all of the scenarios correctly, where as after viewing the video, 64.0% scored the scenarios results were observed after viewing the video for those who used the GCS cards (p = 0.001; RR = 2.0; 95% CI = 1.29 to 3.10) than for those not using the cards (p < 0.0001; RR = 10.0; 95% CI = 2.60 to 38.50). CONCLUSIONS: Post-video viewing scores were better than those observed before the video presentation. Ongoing evaluations include analysis of long-term skill retention and scoring accuracy in the clinical environment. PMID- 12627918 TI - Mass gathering events: retrospective analysis of patient presentations over seven years. AB - INTRODUCTION: St John Ambulance Operations Branch Volunteers have been providing first-aid services at the Royal Adelaide Show for 90 years. The project arose from a need to more accurately predict the workload for first-aid providers at mass gathering events. A formal analysis of workload patterns and the determinants of workload had not been performed. HYPOTHESIS: Casualty presentation workload would be predicted by factors including day of the week, weather, and crowd size. METHOD: Collated and analyzed casualty reports over a seven-year period representing >7,000 patients who presented for first-aid assistance for that period (63 show days) were reviewed retrospectively. RESULTS: Casualty presentations correlated significantly with crowd size, maximum daily temperature, humidity, and day of the week. Patient presentation rate had heterogeneous determinants. The most frequent presentation was minor medical problems with Wednesdays attracting higher casualty presentations and more major medical categories. CONCLUSION: Individual event analysis is a useful mechanism to assist in determining resource allocation at mass gathering events providing an evidence base upon which to make decisions about future needs. Subsequent analysis of other events will assist in supporting accurate predictor models. PMID- 12627920 TI - Ukranian's Disaster Medicine Team Mission to India following the earthquake of 2001. AB - This article describes the basic principles around establishing a Disaster Medicine Camp and the organization of the Ukrainian Disaster Medicine Mobile Hospital, which provided medical aid to victims of the 2001 earthquake in India. All of the information was obtained through direct observation and estimates based on empirical data gathered in the field. PMID- 12627919 TI - Mass-gathering medical care: a review of the literature. AB - Mass-gatherings events provide a difficult setting for which to plan an appropriate emergency medical response. Many of the variables that affect the level and types of medical needs, have not been fully researched. This review examines these variables. METHODS: An extensive review was conducted using the computerized databases Medline and Healthstar from 1977 through May 2002. Articles selected contained information pertaining to mass-gathering variables. These articles were read, abstracted, analyzed, and compiled. RESULTS: Multiple variables are present during a mass gathering, and they interact in complex and dynamic ways. The interaction of these variables contributes to the number of patients treated at an event (medical usage rate) as well as the observed injury patterns. Important variables include weather, event type, event duration, age, crowd mood and density, attendance, and alcohol and drug use. CONCLUSIONS: Developing an understanding of the variables associated with mass gatherings should be the first step for event planners. After these variables are considered, a thorough needs analysis can be performed and resource allocation can be based on objective data. PMID- 12627921 TI - Prehospital interventions: on-scene-time and ambulance-technicians' experience. AB - INTRODUCTION: Very little evidence is available on the experience of ambulance personnels or on the impact of prehospital interventions on total prehospital time. HYPOTHESIS: On-scene-time increases with the number of prehospital techniques used, and ambulance-technicians achieve only limited clinical experience in prehospital techniques. METHODS: Prospective, observational registry study including 56 ambulance technicians from two ambulance stations in the mixed urban/rural county and 5,557 patients who were brought to a hospital in 1998. The number of cases in which each ambulance-technician performed various kinds of prehospital techniques, and the average on-scene time for each prehospital technique and several combinations of prehospital techniques were calculated. RESULTS: There were large differences between the number of times each technique was used. On-scene time was smallest when no techniques were used and tended to increase with the number of interventions used. On-scene-time was relatively low for patients with cardiac arrest. CONCLUSION: The Danish ambulance technicians' curriculum includes interventions for which the technicians only achieve limited practical experience. Prehospital interventions are associated with an increase of on-scene time. PMID- 12627922 TI - Disasters and development: Part 2: understanding and exploiting disaster development linkages. AB - Disasters can impede the effectiveness of development resource allocation. The damage sustained from an event can be classified into four categories: (1) Loss of resources; (2) Interruption of programs and switching of crucial resources to other, shorter-term needs; (3) Negative impacts upon investment climates; and/or (4) Disruption of the non-formal sector (local businesses). Disasters have a particularly destructive economic impact in areas in which there are few alternatives for assets that are destroyed or in areas in which the resources already are at critical levels. Development processes can both increase and/or decrease the vulnerability of a society to hazards. There are dearly established linkages between poverty, marginalization, over-population, and vulnerability. To a large extent, vulnerability derives from poverty. The poor are more likely to live in vulnerable areas (slopes prone to landslides, flood plains, marginal agricultural land), have difficulty accessing education and information, have fewer assets to invest in resources to reduce vulnerability, and are more prone to become malnourished and have chronic illnesses that predispose them to injury and death. Development may be associated with the production of new hazards accepted by a society because the perceived benefits of the development project far exceed the relative risk associated with the project. Therefore, risk assessments must be part of any program planning and evaluation. Training and education are of critical importance in preventing increased vulnerability as a result of development strategies. Development also can progress in a manner that will result in mitigation of the impacts of an event on a given society (increase absorbing capacity and/or buffering capacity, elimination of hazards or the risk of them producing a disaster). Such mitigation measures can be either structural or nonstructural. There exists a wide range of options for incorporating mitigation measures in development projects. Two case studies provide exercises that incorporate the concepts provided in this discussion: (1) The 1985 earthquake in Mexico City; and (2) Agricultural development in northern Sudan. PMID- 12627923 TI - The United States civilian smallpox vaccination program: have we thought through the whole issue? PMID- 12627924 TI - Bioterrorism: pivotal clinical issues. Consensus review of the Society of Infectious Diseases Pharmacists. AB - OBJECTIVES: To discuss specific facts regarding use as a bioweapon, epidemiology, microbiology, clinical manifestations, diagnosis, antimicrobial therapy, immunization, and isolation precautions for five most likely agents of bioterrorism; to review and provide recommendations for health care clinicians on the management of these bioterrorism agents; and to share information on the pharmacist's role in preparedness and response. PARTICIPANTS: The manuscript was drafted by the three authors, reviewed by a group of selected members of the Society of Infectious Diseases Pharmacists, and approved by its Board of Directors. EVIDENCE: The primary focus was to review and summarize recent and key articles on bioterrorism. Preference was given to peer-reviewed journal information and government-sponsored journals, such as the MMWR, Morbidity and Mortality Weekly Report. CONSENSUS PROCESS: Written comments were requested from each reviewer. Comments were incorporated into the final draft. CONCLUSION: Pharmacists play an integral role in disaster preparedness and response and should be involved in planning committees. As drug information specialists, pharmacists can assist other health care providers and emergency personnel, as well as provide counseling to calm, comfort, and empower the public. PMID- 12627925 TI - Integration of population pharmacokinetics, a pharmacodynamic target, and microbiologic surveillance data to generate a rational empiric dosing strategy for cefepime against Pseudomonas aeruginosa. AB - STUDY OBJECTIVE: To derive steady-state pharmacokinetic profiles of cefepime against Pseudomonas aeruginosa. DESIGN: Retrospective evaluation using a weighted approach based on a minimum inhibitory concentration distribution of cefepime in the United States. SETTING: Medical and surgical intensive care units. PATIENTS: One thousand patients with creatinine clearances of 120, 90, or 60 ml/minute. INTERVENTION: Administration of a standard dosage of cefepime 2 g every 12 hours, each dose infused over 0.5 hour, and maximum dosage of 2 g every 8 hours, each dose infused over 0.5 hour; and a nonstandard dosage of 2 g every 12 hours, each dose infused over 6 hours, and continuous infusion of 4 g infused over 24 hours. MEASUREMENTS AND MAIN RESULTS: The standard and maximum dosages achieved pharmacodynamic targets from 4-38% and 21-68%, respectively, for the three groups. With extended infusion of the standard dosage, the probability of achieving the pharmacodynamic target increased to 18-63%. Continuous infusion over 24 hours offered the most promising pharmacodynamic target, attaining 65-81% (p<0.001). CONCLUSION: The recommended dosage of cefepime has a low probability of achieving a pharmacodynamic target predicting a favorable outcome for infections due to P. aeruginosa. The probability of attaining the target could be improved with higher dosages or extended infusion time. PMID- 12627926 TI - Effects of intravenous magnesium sulfate on the QT interval in patients receiving ibutilide. AB - STUDY OBJECTIVE: To determine the effect of intravenous magnesium sulfate on the QT and QTc intervals in patients receiving ibutilide for immediate chemical cardioversion of atrial flutter or fibrillation. DESIGN: Prospective, randomized, double-blind, placebo-controlled trial. SETTING: Hospital cardiology unit. PATIENTS: Twenty patients (mean age 72 +/- 14 yrs) with atrial fibrillation (12 patients) or atrial flutter (8 patients) who were scheduled to receive ibutilide. INTERVENTION: After determining that the patients' baseline QTc intervals were less than 440 msec and baseline serum magnesium levels were within normal limits (mean 2.1 +/- 0.29 mg/dl), the patients were randomly assigned to receive either a 10-minute infusion of magnesium sulfate 2 g in 50 ml of 0.9% sodium chloride or placebo immediately before ibutilide therapy. An additional 2 g of intravenous magnesium sulfate or placebo was given over 1 hour, 10 minutes after the first dose of ibutilide. MEASUREMENTS AND MAIN RESULTS: QT interval duration was measured manually in all 12 leads by using a 0.5-mm-scale precision ruler and magnifying lens. The QT interval increased 29% from baseline at 30 minutes after ibutilide therapy in the placebo group (p=0.007), but no significant change from baseline occurred in the magnesium sulfate group. The 30-minute QTc interval in the placebo group was 18% higher than the baseline value (p=0.01) but did not change significantly in the magnesium sulfate group. QTc changes from baseline were greater in the placebo group than in the magnesium sulfate group at 30 minutes (75 vs 19 msec, respectively, p=0.04). Magnesium sulfate reduced the risk of an ibutilide-induced QTc interval increase of greater than 30 msec or greater than 60 msec and reduced the risk of a QTc interval value of more than 500 msec by 65%, 60%, and 68%, respectively (p=0.07, p=0.175, and p=0.160). CONCLUSIONS: Prophylactic administration of intravenous magnesium sulfate prevents increases in the QT and QTc interval 30 minutes after the last infusion of ibutilide. PMID- 12627927 TI - Shifting from inpatient to outpatient treatment of deep vein thrombosis in a tertiary care center: a cost-minimization analysis. AB - STUDY OBJECTIVE: To compare the cost of contemporary outpatient and historical inpatient management of proximal lower limb deep vein thrombosis (DVT) in adults. DESIGN: Prospective, observational study with historical inpatient cases as controls. SETTING: Ambulatory thrombosis clinic of a tertiary care teaching center in Canada. PATIENTS: Forty-nine inpatients with DVT from a previous study in 1996 at the same institution who would have been eligible for outpatient therapy if this option had been available, and 51 consecutive patients referred to the ambulatory thrombosis clinic for treatment of DVT between March 2000 and January 2001. INTERVENTION: The 49 inpatients received unfractionated heparin, and the 51 outpatients received low-molecular-weight heparin (LMWH). MEASUREMENTS AND MAIN RESULTS: A cost-minimization analysis restricted to the hospital perspective was conducted. This design was justified based on the clinical equivalence of the two treatment strategies. All direct hospital costs for treating the 51 consecutive outpatients with LMWH were measured. These data were compared with the cost of treating the inpatients with unfractionated heparin. The analysis horizon was limited to 7 days, based on the duration of hospitalization and length of heparin therapy for DVT before conversion to oral warfarin. The mean cost (in Canadian dollars) per outpatient case was 248 Canadian dollars (95% confidence interval 216-280 Canadian dollars) and was significantly different from the mean cost/inpatient case of 2826 Canadian dollars (adjusted for the difference in fiscal years) (p<0.0005). A breakdown of the outpatient cost showed that nursing time contributed to 51% of the cost, monitoring laboratory tests 5%, drugs 2%, and other costs (diagnostic laboratory tests and medical imaging) 42%. CONCLUSION: Converting from inpatient to outpatient treatment of proximal DVT was associated with a significant cost savings for our institution. Accordingly, it is financially advantageous for hospitals to offer this service as it reduces direct costs and does not appear to compromise patient care. PMID- 12627929 TI - Increased frequency of venous thromboembolism with the combination of docetaxel and thalidomide in patients with metastatic androgen-independent prostate cancer. AB - STUDY OBJECTIVE: To evaluate the frequency of venous thromboembolism (VTE) in patients with advanced androgen-independent prostate cancer who were treated with docetaxel alone or in combination with thalidomide. DESIGN: Retrospective analysis of a randomized phase II trial. SETTING: National Institutes of Health clinical research center. PATIENTS: Seventy men, aged 50-80 years, with advanced androgen-independent prostate cancer. INTERVENTION: Each patient received either intravenous docetaxel 30 mg/m2/week for 3 consecutive weeks, followed by 1 week off, or the combination of continuous oral thalidomide 200 mg every evening plus the same docetaxel regimen. This 4-week cycle was repeated until there was evidence of excessive toxicity or disease progression. MEASUREMENTS AND MAIN RESULTS: None of 23 patients who received docetaxel alone developed VTE, whereas 9 of 47 patients (19%) who received docetaxel plus thalidomide developed VTE (p=0.025). CONCLUSION: The addition of thalidomide to docetaxel in the treatment of prostate cancer significantly increases the frequency of VTE. Clinicians should be aware of this potential complication when adding thalidomide to chemotherapeutic regimens. PMID- 12627928 TI - Disposition of cefepime in the central nervous system of patients with external ventricular drains. AB - STUDY OBJECTIVE: To assess central nervous system (CNS) penetration of cefepime in adults with external ventricular drains and to compare the achieved cerebrospinal fluid (CSF) concentrations with the usual minimum inhibitory concentrations (MICs) of common pathogens. DESIGN: Open-label, prospective study. SETTING: University-affiliated medical center. PATIENTS: Seven patients with external ventricular drains and normal renal function (documented creatinine clearance > 60 ml/min) who received cefepime 2 g intravenously every 12 hours for treatment of nosocomial pneumonia. INTERVENTION: Serial serum and CSF samples were obtained concurrently after the fourth dose during one dosing interval. MEASUREMENTS AND MAIN RESULTS: The concentration-time profiles in serum and CSF were comodeled by using a two-compartment model with zero-order infusion to the central compartment. The CSF concentration-time profiles of the individual patients were compared with published MIC90 of common pathogens isolated in nosocomial meningitis. Our model reasonably characterized the disposition of cefepime in serum and CSF. Penetration into the CNS was 4-34% based on area under the curve and was 5-58% based on minimum concentration. CONCLUSION: Penetration of cefepime into the CNS was variable among the patients (4-34%) but appeared similar to that reported for other cephalosporins given to treat meningitis. The concentrations attained by most patients in this study were above the MIC90 of many common nosocomial organisms. PMID- 12627930 TI - Important effect of food on the bioavailability of oral testosterone undecanoate. AB - STUDY OBJECTIVE: To assess the effects of food on the bioavailability of testosterone undecanoate, testosterone, and 5alpha-dihydrotestosterone (DHT) after administration of a new oral testosterone undecanoate formulation, Andriol Testocaps. DESIGN: Randomized, open-label, crossover study with a 1-week washout period. SETTING: Clinical pharmacology unit. SUBJECTS: Sixteen healthy postmenopausal women. INTERVENTION: Single oral doses of testosterone undecanoate 80 mg were administered either during a fasting period or after consumption of a standardized continental breakfast. MEASUREMENTS AND MAIN RESULTS: Serum concentrations of testosterone undecanoate were assayed by liquid chromatography with mass spectrometry detection; testosterone and DHT were assayed by gas chromatography with mass spectrometry detection. Serum concentrations of testosterone, testosterone undecanoate, and DHT were low to negligible when testosterone undecanoate was administered to subjects in a fasting state; these values were significantly higher when the test drug was coadministered with food. For testosterone, the maximum serum concentration and area under the plasma concentration-time curve were 0.67 ng/ml and 5.37 ng x hr/ml, respectively, in the fasting state, versus 10.7 ng/ml and 56.4 ng x hr/ml, respectively, in the fed state. The same parameters were also significantly higher for testosterone undecanoate and DHT in the fed versus fasting subjects. CONCLUSION: Food increases the bioavailability of testosterone undecanoate, testosterone, and DHT. For proper absorption, Andriol Testocaps must be taken with meals. PMID- 12627931 TI - Association between adherence to diuretic therapy and health care utilization in patients with heart failure. AB - STUDY OBJECTIVE: To determine the relationship between adherence to diuretic therapy and health care utilization. DESIGN: Prospective, observational study. SETTING: University-affiliated medical center. PATIENTS: Forty-two patients with heart failure. INTERVENTION: Electronic monitoring of adherence to diuretic therapy (percentage of diuretic prescription container openings) and to scheduling (percentage of container openings within a specific time). MEASUREMENTS AND MAIN RESULTS: All patients were prescribed a diuretic, most commonly furosemide (88%). Patients varied widely in adherence to therapy (mu = 72% +/- 30%) and to scheduling (mu = 43% +/- 30%). Education was a predictor of drug-taking adherence (p=0.0062) but not of scheduling adherence. Log-linear models revealed that poor scheduling adherence was associated with increased cardiovascular-related hospitalizations (chi2 11.63, p=0.0006) and predicted more heart failure-related hospitalizations (chi2 4.04, p=0.0444). In contrast, neither measure was significantly associated with cardiovascular- or heart failure-related emergency department visits. We found a moderate correlation between scheduling adherence and taking adherence (r = 0.6513). CONCLUSION: Patients taking a greater proportion of diuretic agents on schedule may decrease the risk of cardiovascular- and heart failure-related hospitalizations. If these findings are confirmed by a larger study, interventions to improve adherence and patient health outcomes should consider the timing of doses as well as the number of daily doses of a diuretic. PMID- 12627932 TI - Effect of levofloxacin coadministration on the international normalized ratios during warfarin therapy. AB - STUDY OBJECTIVE: To evaluate the effect of levofloxacin coadministration on the international normalized ratio (INR) in patients receiving warfarin therapy. DESIGN: Prospective analysis. SETTING: Outpatient clinic at a Veterans Affairs medical center. PATIENTS: Eighteen adult patients receiving warfarin. INTERVENTION: On the basis of clinical diagnosis and judgment, levofloxacin was prescribed to the 18 patients for treatment of various types of infection. The INR was measured before and at 2-8-day intervals after the coadministration of levofloxacin therapy, and once after completing therapy. Warfarin dosages were adjusted when necessary. MEASUREMENTS AND MAIN RESULTS: Warfarin dosages were changed in seven patients as a result of the first nontherapeutic INR values obtained after start of levofloxacin therapy. Owing to a concern regarding noncompliance and the adverse effect of bleeding, warfarin dosage was adjusted in one patient even though his first INR value was in the high end of the therapeutic range (2.98, therapeutic range 2-3). One patient withdrew from the study after the first INR measurement after levofloxacin coadministration. Because of a concern about the possible bleeding complication, warfarin dosage was also adjusted in this patient after obtaining his first INR value. Therefore, only the INR values obtained before and the first INR values obtained after levofloxacin administration were compared to evaluate the effect of levofloxacin on INR determination of warfarin therapy. The INR values obtained before levofloxacin administration did not differ significantly from the first INR values obtained after levofloxacin coadministration (mean +/- SD 2.61 +/- 0.44 vs 2.74 +/- 0.83, 95% confidence interval -0.449-0.196, p=0.419). CONCLUSION: The INR values measured before and after concomitant levofloxacin therapy were not significantly different. However, the ability to detect a significant difference may be affected by the small number of patients studied. Further studies with a larger sample are required to better determine the effect of levofloxacin coadministration on INR monitoring during warfarin therapy PMID- 12627933 TI - Pleconaril, a novel antipicornaviral agent. AB - Despite the availability of therapy for selected symptoms, no specific antiviral agents are available to treat or prevent infections due to the viruses of the Picornaviridae family--rhinoviruses and enteroviruses. Characterization of the three-dimensional structure of picornaviruses in the 1980s allowed development of compounds targeted at the virus itself. Pleconaril is a novel, orally available, systemically acting molecule whose pharmacokinetics are characterized by a two compartment open model with first-order absorption and with a safety profile similar to that of placebo. It shows promising results in treatment of picornaviral respiratory tract infections, meningitis, and other life-threatening infections. PMID- 12627934 TI - Health care resource utilization associated with treatment of penicillin susceptible and -nonsusceptible isolates of Streptococcus pneumoniae. AB - Despite failure to correlate in vitro susceptibility with clinical outcomes for respiratory tract infections and bacteremia, resistance affects management of patients with pneumococcal infections. The economic impact of resistance among pneumococci has not been evaluated. We conducted a single-center, retrospective, observational, cohort study of hospitalized patients infected with Streptococcus pneumoniae isolated from blood or a respiratory source between January 1, 1995, and December 31, 1998. Data were collected for 36 days surrounding the day that the first positive culture was collected. Patients were grouped according to isolate penicillin-susceptibility profile [susceptible minimum inhibitory concentration (MIC) < or = 0.06 microg/ml, nonsusceptible MIC > or = 0.125 microg/ml), and data were analyzed with respect to health care resource utilization patterns. Of 231 patients identified, 142 and 89 had susceptible and nonsusceptible isolates, respectively. Groups were similar with respect to demographics and comorbidities, except that patients infected with a nonsusceptible isolate were more likely to have the isolate obtained from a respiratory source and to have a history of recent antibiotic therapy. No difference was noted with respect to clinical outcome; however, patients infected with a nonsusceptible isolate had a longer median stay (14 vs 10 days, p<0.05). They also had significantly higher total median costs (1600 dollars, 95% confidence interval 257-2943 dollars) due to room and nursing services. Infections caused by penicillin-nonsusceptible pneumococci were not associated with a worse outcome in hospitalized patients but were associated with increased cost of care. PMID- 12627936 TI - Retrospective review of neurotoxicity induced by cefepime and ceftazidime. AB - We reviewed 42 cases of cefepime-induced neurotoxicity and 12 cases of ceftazidime-induced neurotoxicity from the literature and our institution. Clinical characteristics and timing of diagnosis were examined. Common findings were confusion with temporospatial disorientation (96% of patients), myoclonus (33%), and seizures (13%). These neurologic disorders frequently are encountered in uremic and elderly patients, who often are in a confused state when they visit their physician. The risk of delayed diagnosis was greater with cefepime than ceftazidime neurotoxicity. The median interval between symptom onset and diagnosis of cefepime versus ceftazidime neurotoxicity was 5 and 3 days, respectively (p=0.005). Delayed diagnosis of cefepime neurotoxicity may be due to lack of awareness of the adverse effect. Data gathered since these two broad spectrum antibiotics were first marketed underscore the potential for neurologic adverse events secondary to their administration. Thus, clinicians' awareness must be increased so that the time between symptom onset and diagnosis can be reduced. PMID- 12627935 TI - Evaluation of the clinical and economic impact of a brand name-to-generic warfarin sodium conversion program. AB - BACKGROUND: Substitution of generic warfarin initially was discouraged because of concerns regarding therapeutic failure or toxicity. Although subsequent research with AB-rated (i.e., bioequivalent) warfarin did not confirm initial concerns, the issue is not settled for all clinicians. OBJECTIVES: We sought to provide additional information regarding the clinical and economic impact of warfarin conversion by analyzing a real-life sample of patients receiving long-term anticoagulation therapy who were switched from brand name to generic warfarin. METHODS: Patients who had been taking warfarin for at least 180 days and had received uninterrupted oral anticoagulation 90 days before and 90 days after switching to generic warfarin were included. The switch date was based on the first time generic warfarin was dispensed from our pharmacies. The primary end point was the calculated amount of time each patient's international normalized ratio (INR) values were within the patient-specific target INR range in the 90 days before and after the switch. Data regarding adverse events and medical resource utilization were also collected. Pharmacoeconomic analyses were performed. RESULTS: The analysis included 2299 patients. The overall difference in calculated time INR values were below (22.6% before vs 26.1% after switch, p<0.0001) and within (65.9% before vs 63.3% after switch, p=0.0002) the therapeutic INR range was statistically but not clinically significant. Only 28.0% of patients experienced a change in therapeutic INR control of 10% or less, 33.1% experienced INR control that improved by greater than 10%, and 38.9% experienced INR control that worsened by more than 10%. The difference in total treatment costs associated with brand name and generic warfarin was 3128 dollars/100 patient-years in favor of the generic product. Sensitivity analyses revealed that cost savings associated with warfarin conversion in this health care system were highly dependent on the difference between warfarin costs and cost of treating anticoagulation-related adverse events. CONCLUSIONS: Most of these patients were successfully switched from brand name to generic warfarin. However, supplemental INR monitoring is warranted when one warfarin product is substituted for another to allow timely detection of those patients who experience significant changes in anticoagulation response. PMID- 12627937 TI - Acute profound thrombocytopenia associated with eptifibatide therapy. AB - An 80-year-old woman and a 79-year-old man underwent urgent percutaneous coronary intervention and received adjunctive eptifibatide. Platelet counts in both patients fell to below 20 x 10(3)/mm3 within 4 hours of eptifibatide administration. Reports in the medical literature reinforce the importance of recognizing that eptifibatide can cause acute profound thrombocytopenia. All three available glycoprotein IIb-IIIa inhibitors--abciximab, eptifibatide, and tirofiban--have been associated with the development of this disorder. Thus, clinicians should routinely monitor platelet counts in patients receiving glycoprotein IIb-IIIa inhibitors within 2-4 hours of the start of the infusion. PMID- 12627938 TI - Possible case of mania associated with ma-huang. AB - Several commercially available weight-loss supplements contain ma-huang, an herb derived from Ephedra sinica. Previous reports have raised awareness that ma-huang supplements may precipitate symptoms consistent with mania in susceptible individuals. A 21-year-old woman required psychiatric hospitalization as a result of acute manic symptoms with psychosis. The emergence of her symptoms coincided with her use of two supplements containing ma-huang. Consumers need to be educated about the potential adverse psychiatric effects of ma-huang in order to make well-informed decisions before using such supplements. A preexisting psychiatric disorder may also increase susceptibility to these adverse effects. PMID- 12627939 TI - 20 years since human immunodeficiency virus discovery: considerations for the next decade. AB - The year 2003 marks the 20th anniversary of the discovery of the human immunodeficiency virus type 1 (HIV-1). Among infectious disease-causing agents, HIV-1 is now the number one killer worldwide. Approximately 70% of the cases in the world are in sub-Saharan Africa, where in some regions, the seroprevalence of HIV-1 among adults exceeds 25%. Its high seroprevalence in some countries has raised concern that acquired immunodeficiency syndrome may create a threat to world peace. Despite developments in molecular biology, virology, immunology, and pharmacology, control of HIV-1 still awaits effective vaccines and microbicides. Because significant technologic advances still are needed to overcome the obstacles posed by HIV-1, we must find ways to expand and expedite proven prevention strategies and provide access to HIV-1 treatment for infected individuals throughout the world. Without doing so, the worst of this global pandemic will occur in the next decade before effective vaccines and microbicides are available. PMID- 12627941 TI - Metal binding and oxidation of amyloid-beta within isolated senile plaque cores: Raman microscopic evidence. AB - Alzheimer's disease (AD) is characterized by the deposition of amyloid plaques in the parenchyma and vasculature of the brain. Although previous analytical studies have provided much information about the composition and structure of synthetic amyloid-beta fibrils, there is, surprisingly, a dearth of data on intact amyloid plaques from AD brain. Therefore, to elucidate the structure and detailed composition of isolated amyloid plaque cores, we utilized a high-resolution, nondestructive technique, Raman microscopy. The data are of very high quality and contain detailed information about protein composition and conformation, about post-translational modification, and about the chemistry of metal binding sites. Remarkably, spectra obtained for senile plaque (SP) cores isolated from AD brain are essentially identical both within and among brains. The Raman data show for the first time that the SP cores are composed largely of amyloid-beta and confirm inferences from X-ray studies that the structure is beta-sheet with the additional possibility that this may be present as a parallel beta-helix. Raman bands characteristic of methionine sulfoxide show that extensive methionine oxidation has occurred in the intact plaques. The Raman spectra also demonstrate that Zn(II) and Cu(II) are coordinated to histidine residues in the SP cores, at the side chains' N(tau) and N(pi) atoms, respectively. Treatment of the senile plaques with the chelator ethylenediaminetetraacetate reverses Cu binding to SP histidines and leads to a broadening of amide features, indicating a "loosening" of the beta-structure. Our results indicate that Abeta in vivo is a metalloprotein, and the loosening of the structure following chelation treatment suggests a possible means for the solubilization of amyloid deposits. The results also reveal a direct chemical basis for oxidative damage caused by amyloid-beta protein in AD. PMID- 12627940 TI - Sequence analyses of G-protein-coupled receptors: similarities to rhodopsin. PMID- 12627942 TI - Facile detection of protein-protein interactions by one-dimensional NMR spectroscopy. AB - Two methods for detecting protein-protein interactions in solution using one dimensional (1D) NMR spectroscopy are described. Both methods rely on measurement of the intensity of the strongest methyl resonance (SMR), which for most proteins is observed at 0.8-0.9 ppm. The severe resonance overlap in this region facilitates detection of the SMR at low micromolar and even sub-micromolar protein concentrations. A decreased SMR intensity in the 1H NMR spectrum of a protein mixture compared to the added SMR intensities of the isolated proteins reports that the proteins interact (SMR method). Decreased SMR intensities in 1D 13C-edited 1H NMR spectra of 13C-labeled proteins upon addition of unlabeled proteins or macromolecules also demonstrate binding (SMRC method). Analysis of the interaction between XIAP and Smac, two proteins involved in apoptosis, illustrates both methods. A study showing that phospholipids compete with the neuronal core complex for Ca2+-dependent binding to the presynaptic Ca2+-sensor synaptotagmin 1 illustrates the usefulness of the SMRC method in studying multicomponent systems. PMID- 12627943 TI - Catalytic and structural effects of amino acid substitution at histidine 30 in human manganese superoxide dismutase: insertion of valine C gamma into the substrate access channel. AB - Catalysis of the disproportionation of superoxide by human manganese superoxide dismutase (MnSOD) is characterized by an initial burst of catalysis followed by a much slower region that is zero order in superoxide and due to a product inhibition by peroxide anion. We have prepared site-specific mutants with replacements at His30, the side chain of which lies along the substrate access channel and is about 5.8 A from the metal. Using pulse radiolysis to generate superoxide, we have determined that kcat/K(m) was decreased and product inhibition increased for H30V MnSOD, both by 1-2 orders of magnitude, compared with wild type, H30N, and H30Q MnSOD. These effects are not attributed to the redox potentials, which are similar for all of these variants. An investigation of the crystal structure of H30V Mn(III)SOD compared with wild type, H30Q, and H30N Mn(III)SOD showed the positions of two gamma carbons of Val30 in the active site; Cgamma1 overlaps Cgamma of His30 in wild type, and Cgamma2 extends into the substrate access channel and occupies the approximate position of a water molecule in the wild type. The data suggest that Cgamma2 of the Val side chain has significantly interrupted catalysis by this overlap into the access channel with possible overlap with the substrate-product binding site. This is supported by comparison of the crystal structure of H30V MnSOD with that of azide bound to Mn(III)SOD from Thermus thermophilus and by visible absorption spectra showing that azide binding to the metal in H30V Mn(III)SOD is abolished. Moreover, the presence of Val30 caused a 100-fold decrease in the rate constant for dissociation of the product-inhibited complex compared with wild type. PMID- 12627944 TI - Proton transfer reactions in the F86D and F86E mutants of pharaonis phoborhodopsin (sensory rhodopsin II). AB - pharaonis phoborhodopsin (ppR, also called pharaonis sensory rhodopsin II, psRII), a negative phototaxis receptor of Natronobacterium pharaonis, can use light to pump a proton in the absence of its transducer protein. However, the pump activity is much lower than that of the light-driven proton-pump bacteriorhodopsin (BR). ppR's pump activity is known to be increased in a mutant protein, in which Phe86 is replaced with Asp (F86D). Phe86 is the amino acid residue corresponding to Asp96 in BR, and we expect that Asp86 plays an important role in the proton transfer at the highly hydrophobic cytoplasmic domain of the F86D mutant ppR. In this article, we studied protein structural changes and proton transfer reactions during the photocycles of the F86D and F86E mutants in ppR by means of Fourier transform infrared (FTIR) spectroscopy and photoelectrochemical measurements using a tin oxide (SnO2) electrode. FTIR spectra of the unphotolyzed state and the K and M intermediates are very similar among F86D, F86E, and the wild type. Asp86 or Glu86 is protonated in F86D or F86E, respectively, and the pK(a) > 9. During the photocycle, the pK(a) is lowered and deprotonation of Asp86 or Glu86 is observed. Detection of both deprotonation of Asp86 or Glu86 and concomitant reprotonation of the 13-cis chromophore implies the presence of a proton channel between position 86 and the Schiff base. However, the photoelectrochemical measurements revealed proton release presumably from Asp86 or Glu86 to the cytoplasmic aqueous phase in the M state. This indicates that the ppR mutants do not have the BR-like mechanism that conducts a proton uniquely from Asp86 or Glu86 (Asp96 in BR) to the Schiff base, which is possible in BR by stepwise protein structural changes at the cytoplasmic side. In ppR, there is a single open structure at the cytoplasmic side (the M like structure), which is shown by the lack of the N-like protein structure even in F86D and F86E at alkaline pH. Therefore, it is likely that a proton can be conducted in either direction, the Schiff base or the bulk, in the open M-like structure of F86D and F86E. PMID- 12627945 TI - Role of electrostatic interactions in PDZ domain ligand recognition. AB - PDZ domains are protein-protein interaction modules that normally recognize short C-terminal peptides. The apparent requirement for a ligand with a free terminal carboxylate group has led to the proposal that electrostatic interactions with the terminus play a significant role in recognition. However, this model has been called into question by the more recent finding that PDZ domains can recognize some internal peptide motifs that occur within a specific secondary structure context. Although these motifs bind at the same interface, they lack a terminal charge. Here we have investigated the role of electrostatics in PDZ-mediated recognition in the mouse alpha1-syntrophin PDZ domain by examining the salt dependence of binding to both terminal and internal ligands and the effects of mutating a conserved basic residue previously proposed to play a role in electrostatic recognition. These studies indicate that direct electrostatic interactions with the peptide terminus do not play a significant energetic role in binding. Additional chemical modification studies of the peptide terminus support a model in which steric and hydrogen bonding complementarity play a primary role in recognition specificity. Peptides with a free carboxy terminus, or presented within a specific structural context, can satisfy these requirements. PMID- 12627946 TI - A flavodiiron protein and high molecular weight rubredoxin from Moorella thermoacetica with nitric oxide reductase activity. AB - A five-gene "oxidative stress protection" cluster has recently been described from the strictly anaerobic, acetogenic bacterium, Moorella thermoacetica [Das, A., et al. (2001) J. Bacteriol. 183, 1560-1567]. Within this cluster are two cotranscribed genes, fprA (for A-type flavoprotein) and hrb (for high molecular weight rubredoxin) whose encoded proteins have no known functions. Here we show that FprA and Hrb are expressed in M. thermoacetica under normal anaerobic growth conditions and report characterizations of the recombinant FprA and Hrb. FprA contains flavin mononucleotide (FMN) and a non-heme diiron site. Mossbauer spectroscopy shows that the irons of the diferric site are antiferromagnetically coupled, implying a single-atom, presumably solvent, bridge between the irons. Hrb contains FMN and a rubredoxin-like [Fe(SCys)4] site. NADH does not directly reduce either the FMN or the diiron site in FprA, whereas Hrb functions as an efficient NADH:FprA oxidoreductase. Substitution of zinc for iron in Hrb completely abolished this activity. The observation that homologues of FprA from other organisms show O2 and/or anaerobic NO consumption activity prompted an examination of these activities for M. thermoacetica FprA. The Hrb/FprA combination does indeed have both NADH:O2 and NADH:NO oxidoreductase activities. The NO reductase activity, however, was significantly more efficient due to a lower Km for NO (4 M) and to progressive and irreversible inactivation of FprA during O2 reductase turnover but retention of activity during NO reductase turnover. Substitution of zinc for iron in FprA completely abolished these reductase activities. The stoichiometry of 1 mol of NADH oxidized:2 mol of NO consumed implies reduction to N2O. Fits of an appropriate rate law to the kinetics data are consistent with a mechanism in which 2NO's react at each FprA active site in the committed step. Expression of FprA in an Escherichia coli strain deficient in NO reductase restored the anaerobic growth phenotype of cultures exposed to otherwise toxic levels of exogenous NO. The accumulated results indicate that Hrb/FprA is fully capable of functioning in nitrosative stress protection in M. thermoacetica. PMID- 12627947 TI - Design of a ruthenium-labeled cytochrome c derivative to study electron transfer with the cytochrome bc1 complex. AB - A new ruthenium-cytochrome c derivative was designed to study electron transfer from cytochrome bc1 to cytochrome c (Cc). The single sulfhydryl on yeast H39C;C102T iso-1-Cc was labeled with Ru(2,2'-bipyrazine)2(4-bromomethyl-4'-methyl 2,2'-bipyridine) to form Ru(z)-39-Cc. The Ru(z)-39-Cc derivative has the same steady-state activity with yeast cytochrome bc1 as wild-type yeast iso-1-Cc, indicating that the ruthenium complex does not interfere in the binding interaction. Laser excitation of reduced Ru(z)-39-Cc results in electron transfer from heme c to the excited state of ruthenium with a rate constant of 1.5 x 10(6) x s(-1). The resulting Ru(I) is rapidly oxidized by atmospheric oxygen in the buffer. The yield of photooxidized heme c is 20% in a single flash. Flash photolysis of a 1:1 complex between reduced yeast cytochrome bc1 and Ru(z)-39-Cc at low ionic strength leads to rapid photooxidation of heme c, followed by intracomplex electron transfer from cytochrome c1 to heme c with a rate constant of 1.4 x 10(4) x s(-1). As the ionic strength is raised above 100 mM, the intracomplex phase disappears, and a new phase appears due to the bimolecular reaction between solution Ru-39-Cc and cytochrome bc1. The interaction of yeast Ru-39-Cc with yeast cytochrome bc1 is stronger than that of horse Ru-39-Cc with bovine cytochrome bc1, suggesting that nonpolar interactions are stronger in the yeast system. PMID- 12627948 TI - NMR solution structure of the glucagon antagonist [desHis1, desPhe6, Glu9]glucagon amide in the presence of perdeuterated dodecylphosphocholine micelles. AB - Glucagon, a 29-residue peptide hormone, plays an important role in glucose homeostasis and in diabetes mellitus. Several glucagon antagonists and agonists have been developed, but limited structural information is available to clarify the basis of their biological activity. The solution structure of the potent glucagon antagonist, [desHis1, desPhe6, Glu9]glucagon amide, was determined by homonuclear 2D NMR spectroscopy at pH 6.0 and 37 degrees C in perdeuterated dodecylphosphocholine micelles. The overall backbone root-mean-square deviation (rmsd) for the structured portion (residues 7-29, glucagon numbering) of the micelle-bound 27-residue peptide is 1.36 A for the 15 lowest-energy structures, after restrained molecular dynamics simulation. The structure consists of four regions (segment backbone rmsd in A): an unstructured N-terminal segment between residues 2 and 5 (1.68), an irregular helix between residues 7 and 14 (0.79), a hinge region between residues 15 and 18 (0.54), and a well-defined alpha-helix between residues 19 and 29 (0.33). The two helices form an L-shaped structure with an angle of about 90 degrees between the helix axes. There is an extended hydrophobic cluster, which runs along the inner surface of the L-structure and incorporates the side chains of the hydrophobic residues of each of the amphipathic helices. The outer surface contains the hydrophilic side chains, with two salt bridges (D15-R18 and R17-D21) implied from close approach of the charged groups. This result is the first clear indication of an overall tertiary fold for a glucagon analogue in the micelle-bound state. The relationship of the two helical structural elements may have important implications for the biological activity of the glucagon antagonist. PMID- 12627949 TI - Anticarcinogenic Bowman Birk inhibitor isolated from snail medic seeds (Medicago scutellata): solution structure and analysis of self-association behavior. AB - The high-resolution three-dimensional structure of a Bowman Birk inhibitor, purified from snail medic seeds (Medicago scutellata) (MSTI), has been determined in solution by 1H NMR spectroscopy at pH 5.6 and 27 degrees C. The structure of MSTI comprises two distinct symmetric domains each composed of a three-stranded beta-sheet containing a VIb type loop, where the active sites are located. A characteristic geometry of three aromatic residues confers stability to this protein, and we observe that this feature is conserved in all the Bowman Birk inhibitors of known structure. The two active domains exhibit different conformational features: the second domain displays higher flexibility and hydrophobicity with respect to the first one, and these properties have been correlated to a lower trypsin inhibitory specificity, in agreement with titration studies that have shown a stoichiometric ratio MSTI:trypsin of 1:1.5. NMR analysis indicated that MSTI undergoes self-association at concentrations higher than 2 mM, and the residues involved in this mechanism are localized at opposite faces of the molecule, having the highest positive and negative potential, respectively, thus indicating that electrostatic intermolecular interactions are the driving forces for MSTI association. Most of the residues affected by self association are highly conserved in BBIs from different seeds, suggesting a functional relevance for these charged superficial patches, possibly involved in the interaction with other enzymes or macromolecules, thus triggering anti carcinogenic activity. PMID- 12627950 TI - NMR determination of pKa values for Asp, Glu, His, and Lys mutants at each variable contiguous enzyme-inhibitor contact position of the turkey ovomucoid third domain. AB - From the larger set of 191 variants at all the variable contact positions in the turkey ovomucoid third domain, we selected a subset that consists of Asp, Glu, His, and Lys residues at eight of the nine contiguous P6-P3' positions (residues 13-21), the exception being P3-Cys16 which is involved in a conserved disulfide bridge. Two-dimensional [1H,1H]-TOCSY data were collected for each variant as a function of sample pH. This allowed for the evaluation of 31 of the 32 pK(a) values for these residues, the exception being that of P5-Lys14, whose signals at high pH could not be resolved from those of other Lys residues in the molecule. Only two of the titrating residues are present in the wild-type protein (P6-Lys13 and P1'-Glu19); hence, these measurements complement earlier measurements by A. D. Robertson and co-workers. This data set was supplemented with results from the pH dependence of NMR spectra of four additional single mutants, P1-Leu18Gly, P1 Leu18Ala, P2-Thr17Val, and P3'-Arg21Ala, and two double mutants, P2-Thr17Val/P3' Arg21Ala and P8-Tyr11Phe/P6-Lys13Asp. Probably the most striking result was observation of a P2-Thr17...P1'-Glu19 hydrogen bond and a P1'-Glu19-P3'-Arg21 electrostatic interaction within the triad of P2, P1', and P3' (residues 17, 19, and 21, respectively). In several cases, the pK(a) of a particular residue was sensed by resonances not only in that residue but also in residue(s) with which it interacts. Remarkably, in several interacting systems, resonances from different protons within the same residue yielded different pHmid values. PMID- 12627951 TI - Crystal structure of ribosomal protein L30e from the extreme thermophile Thermococcus celer: thermal stability and RNA binding. AB - We report here the high-resolution crystal structure of the ribosomal protein L30e from the hyperthermophilic archaeon Thermococcus celer determined at cryo temperature. When it is compared with its mesophilic homologue, L30e from yeast, a number of structural features that can enhance thermostability are revealed. Disordered residues corresponding to a large RNA-binding loop in yeast L30e are well structured in the T. celer protein. The overall charge of T. celer L30e is near neutral, whereas that of the yeast homologue is highly positive. This is the result of an increase in the number of acidic residues at the expense of polar residues, Asn, Ser, and Thr. Extensive ion pair networks are found on the molecular surface. Exposed nonpolar surface areas are reduced in the T. celer protein. Its side chain atoms preferably form hydrogen bonds with main chain atoms. Taken together, these factors contribute to high protein stability. The roles of well-conserved L30e residues are studied and found to be important in defining a very compact overall structure and in maintaining the structure of the RNA binding site. By comparing it with the yeast homologue, we also identified the residues that are responsible for RNA binding and built a model to illustrate how L30e binds to an RNA kink turn motif. PMID- 12627952 TI - The 1.75 A crystal structure of acetyl-CoA synthetase bound to adenosine-5' propylphosphate and coenzyme A. AB - Acetyl-coenzyme A synthetase catalyzes the two-step synthesis of acetyl-CoA from acetate, ATP, and CoA and belongs to a family of adenylate-forming enzymes that generate an acyl-AMP intermediate. This family includes other acyl- and aryl-CoA synthetases, firefly luciferase, and the adenylation domains of the modular nonribosomal peptide synthetases. We have determined the X-ray crystal structure of acetyl-CoA synthetase complexed with adenosine-5'-propylphosphate and CoA. The structure identifies the CoA binding pocket as well as a new conformation for members of this enzyme family in which the approximately 110-residue C-terminal domain exhibits a large rotation compared to structures of peptide synthetase adenylation domains. This domain movement presents a new set of residues to the active site and removes a conserved lysine residue that was previously shown to be important for catalysis of the adenylation half-reaction. Comparison of our structure with kinetic and structural data of closely related enzymes suggests that the members of the adenylate-forming family of enzymes may adopt two different orientations to catalyze the two half-reactions. Additionally, we provide a structural explanation for the recently shown control of enzyme activity by acetylation of an active site lysine. PMID- 12627953 TI - IL-8-induced migratory responses through CXCR1 and CXCR2: association with phosphorylation and cellular redistribution of focal adhesion kinase. AB - CXCR1 and CXCR2 mediate migratory activities in response to IL-8 and other ELR+ CXC chemokines (e.g., GCP-2 and NAP-2). In vitro, activation of migration is induced by low IL-8 concentrations (10-50 ng/mL), whereas migratory shut-off is induced by high IL-8 concentrations (1000 ng/mL). The stimulation of CXCR1 and CXCR2 by IL-8 concentrations that result in migratory activation induced focal adhesion kinase (FAK) phosphorylation in a G(alpha)i-dependent manner. The expression of FRNK, a dominant negative mutant of FAK, perturbed migratory responses to the activating dose of 50 ng/mL IL-8. The migration-activating concentrations of 50 ng/mL GCP-2 and NAP-2 induced less potent migratory responses and FAK phosphorylation in CXCR2-expressing cells as compared with IL 8. These results indicate that FAK is phosphorylated, and required, for the chemotactic response under conditions of migratory activation by ELR+-CXC chemokines. In addition, FAK phosphorylation was determined following exposure to migration-attenuating concentrations of IL-8. In CXCR1-RBL cells this treatment resulted in FAK phosphorylation, in similar levels to those induced by activating concentrations of IL-8. In contrast, in CXCR2-RBL cells the migration-attenuating concentrations of IL-8 induced promoted levels of FAK phosphorylation and different patterns of FAK phosphorylation on its six potential tyrosine phosphorylation sites, as compared to activating concentrations of the chemokine. Exposure to IL-8 resulted not only in FAK phosphorylation but also in its cellular redistribution, indicated by the formation of defined contact regions with the substratum, enriched in phosphorylated FAK and vinculin. Overall, FAK phosphorylation was associated with, and found to be differently regulated upon, ELR+-CXC chemokine-induced migration. PMID- 12627954 TI - Regulation of epidermal growth factor receptor internalization by G protein coupled receptors. AB - The epidermal growth factor (EGF) receptor (EGFR) plays a central role in regulating cell proliferation, differentiation, and migration. Cellular responses to EGF are dependent upon the amount of EGFR present on the cell surface. Stimulation with EGF induces sequestration of the receptor from the plasma membrane and its subsequent downregulation. Recently, internalization of the EGFR was also shown to be required for mitogenic signaling via the activation of MAP kinases. Therefore, mechanisms regulating internalization of the EGFR represent an important facet for the control of cellular response. Here, we demonstrate that EGFR is removed from the cell surface not only following stimulation with EGF, but also in response to stimulation of G protein-coupled lysophosphatidic acid (LPA) and beta2 adrenergic (beta2AR) receptors. Using a FLAG epitope-tagged EGFR to quantitate receptor internalization, we show that incubation with EGF, LPA, or isoproterenol (ISO) causes the time-dependent loss of cell surface EGFR. Internalization of EGFR by these ligands involves the tyrosine kinase activity of the receptor itself and c-Src, as well as the GTPase activity of dynamin. Unexpectedly, we find that internalization of the EGFR by EGF is dependent upon Gbetagamma and beta-arrestin proteins; expression of minigenes encoding the carboxyl terminii of the G protein-coupled receptor kinase 2, or beta-arrestin1, attenuates LPA-, ISO-, and EGF-mediated internalization of EGFR. Thus, G protein coupled receptors can control the function of the EGFR by regulating its endocytosis. PMID- 12627955 TI - Catalytic mechanism of the Streptomyces K15 DD-transpeptidase/penicillin-binding protein probed by site-directed mutagenesis and structural analysis. AB - The Streptomyces K15 penicillin-binding DD-transpeptidase is presumed to be involved in peptide cross-linking during bacterial cell wall peptidoglycan assembly. To gain insight into the catalytic mechanism, the roles of residues Lys38, Ser96, and Cys98, belonging to the structural elements defining the active site cleft, have been investigated by site-directed mutagenesis, biochemical studies, and X-ray diffraction analysis. The Lys38His and Ser96Ala mutations almost completely abolished the penicillin binding and severely impaired the transpeptidase activities while the geometry of the active site was essentially the same as in the wild-type enzyme. It is proposed that Lys38 acts as the catalytic base that abstracts a proton from the active serine Ser35 during nucleophilic attack and that Ser96 is a key intermediate in the proton transfer from the Ogamma of Ser35 to the substrate leaving group nitrogen. The role of these two residues should be conserved among penicillin-binding proteins containing the Ser-Xaa-Asn/Cys sequence in motif 2. Conversion of Cys98 into Asn decreased the transpeptidase activity and increased hydrolysis of the thiolester substrate and the acylation rate with most beta-lactam antibiotics. Cys98 is proposed to play the same role as Asn in motif 2 of other penicilloyl serine transferases in properly positioning the substrate for the catalytic process. PMID- 12627956 TI - Amino acid residues in the nicotinamide binding site contribute to catalysis by horse liver alcohol dehydrogenase. AB - Amino acid residues Thr-178, Val-203, and Val-292, which interact with the nicotinamide ring of the coenzyme bound to alcohol dehydrogenase (ADH), may facilitate hydride transfer and hydrogen tunneling by orientation and dynamic effects. The T178S, T178V, V203A, V292A, V292S, and V292T substitutions significantly alter the steady state and transient kinetics of the enzyme. The V292A, V292S, and V292T enzymes have decreased affinity for coenzyme (NAD+ by 30 50-fold and NADH by 35-75-fold) as compared to the wild-type enzyme. The substitutions in the nicotinamide binding site decrease the rate constant of hydride transfer for benzyl alcohol oxidation by 3-fold (for V292T ADH) to 16 fold (for V203A ADH). The modest effects suggest that catalysis does not depend critically on individual residues and that several residues in the nicotinamide binding site contribute to catalysis. The structures of the V292T ADH-NAD+ pyrazole and wild-type ADH-NAD+-4-iodopyrazole ternary complexes are very similar. Only subtle changes in the V292T enzyme cause the large changes in coenzyme binding and the small change in hydride transfer. In these complexes, one pyrazole nitrogen binds to the catalytic zinc, and the other nitrogen forms a partial covalent bond with C4 of the nicotinamide ring, which adopts a boat conformation that is postulated to be relevant for hydride transfer. The results provide an experimental basis for evaluating the contributions of dynamics to hydride transfer. PMID- 12627957 TI - A rugged energy landscape mechanism for trapping of transmembrane receptors during endocytosis. AB - Efficient clathrin-mediated endocytosis of transmembrane receptors requires that clathrin-coated pits retain the receptors long enough to allow vesicle formation and internalization. In many cases, however, the receptors can exhibit mean lifetimes in coated pits much shorter than the lifetime of the pit at the plasma membrane. A rugged energy landscape for binding, which produces a broad distribution of residence times, ensures a significant probability of times much greater than the mean and would allow efficient trapping of proteins in these cases. We used fluorescence correlation spectroscopy and total internal reflection microscopy to measure the kinetics of movement of a C5a receptor yellow fluorescent protein fusion in living cells. These experiments demonstrate that clusters of trapped receptors exhibit fluctuations in fluorescence intensity that vary in time scale over 2 orders of magnitude. Most of the variation in intensity is likely due to the motion of the receptors in the plane of the plasma membrane, although it is not possible to rule out a small contribution from motion orthogonal to the plane of the membrane. The broad time scale distribution of the intensity fluctuations is consistent with a rugged energy landscape mechanism for trapping of the receptors. This mechanism, which allows efficient trapping to coexist with rapid exchange, may also be relevant to other biological processes involving binding in heterogeneous chemical environments. PMID- 12627958 TI - Purification and properties of human blue-light photoreceptor cryptochrome 2. AB - Cryptochromes are blue-light photoreceptors that regulate the circadian clock in animals and growth and development in plants. Cryptochromes have high sequence homology to DNA photolyase but appear to lack photorepair activity. All previous work on cryptochromes was performed with protein expressed in heterologous systems; hence, biochemical and photochemical studies performed with these proteins were subject to certain limitations. In this study, we purified cryptochrome 2 (hCRY2) from human cells and characterized it. We find that hCRY2 exhibits fluorescence properties consistent with the presence of folate and flavin cofactors. Cryptochrome 2 binds to double-stranded DNA weakly and to single-stranded DNA with higher affinity, and this binding is further stimulated by the presence of a (6-4) photoproduct. However, light has no effect on the cryptochrome 2-(6-4) photoproduct complex. These findings reveal new properties of this protein already known to function as a circadian photoreceptor and a light-independent negative transcriptional regulator of the clock genes. PMID- 12627959 TI - Ornithine decarboxylase promotes catalysis by binding the carboxylate in a buried pocket containing phenylalanine 397. AB - Ornithine decarboxylase (ODC) is a pyridoxal 5'-phosphate (PLP) dependent enzyme that catalyzes the decarboxylation of l-Orn to putrescine, a rate-limiting step in the formation of polyamines. The X-ray crystal structures of ODC, complexed to several ligands, support a model where the substrate is oriented with the carboxyl-leaving group buried on the re face of the PLP cofactor. This binding site is composed of hydrophobic and electron-rich residues, in which Phe-397 is predicted to form a close contact. Mutation of Phe-397 to Ala reduces the steady state rate of product formation by 150-fold. Moreover, single turnover analysis demonstrates that the rate of the decarboxylation step is decreased by 2100-fold, causing this step to replace product release as the rate-limiting step in the mutant enzyme. These data support the structural prediction that the carboxyl leaving group is positioned to interact with Phe-397. Multiwavelength stopped flow analysis of reaction intermediates suggests that a major product of the reaction with the mutant enzyme is pyridoximine 5'-phosphate (PMP), resulting from incorrect protonation of the decarboxylated intermediate at the C4' position. This finding was confirmed by HPLC analysis of the reaction products, demonstrating that Phe-397 also plays a role in maintaining the integrity of the reaction chemistry. The finding that the carboxylate-leaving group is oriented on the buried side of the PLP cofactor suggests that ODC facilitates decarboxylation by destabilizing the charged substrate carboxyl group in favor of an electrostatically more neutral transition state. PMID- 12627960 TI - Active site loop motion in triosephosphate isomerase: T-jump relaxation spectroscopy of thermal activation. AB - As for many enzymes, the enzymatic pathway of triosephosphate isomerase (TIM) includes the partially rate determining motion of an active site loop (loop 6, residues 166-176), which must remain closed during chemistry but must open just before product release. The motion of this loop was monitored using laser induced temperature-jump relaxation spectroscopy at nanosecond to millisecond time resolution. Trp168 in the hinge of the mobile loop served as a fluorophore reporter in a mutant of the yeast enzyme. The opening rate was studied as a function of the concentration of glycerol 3-phosphate, a substrate surrogate. Monoexponential kinetics were observed; assuming a simple two-step ligand release mechanism involving an encounter complex intermediate, the time scales of loop opening and closing were derived. The opening rate of the loop at 25 degrees C was determined to be 2500 +/- 1000 s(-1), in remarkable agreement with solution and solid state NMR measurements. The closing rate at the same temperature was 46,700 +/- 1800 s(-1). The rates were also studied as a function of the sample temperature following the jump. Enthalpies of activation of the loop motion, DeltaH(close) and DeltaH(open), were estimated to be 13.8 and 14.1 kcal/mol, respectively. The enthalpy of dissociation estimated from the kinetic studies is in reasonable agreement with steady-state values. Moreover, the enthalpy was dissected, for the first time, into components associated with ion binding and with protein conformational change. The enthalpy of the release reaction appeared to have a substantial contribution from the dissociation of the ligand from the encounter complex, found to be endothermic at 6 kcal/mol. In contrast, the population ratio of the open to closed loop conformations is found to favor the closed conformation but to be substantially less temperature dependent than the release step. Preliminary data of other ligands show that G3P behavior resembles that of the substrate but differs from 2-phosphoglycolate, a tight binding inhibitor, and phosphate. This study represents one of the first detailed comparisons between NMR and fluorescence based probes of protein motion and results in good agreement between the methods. The data in aggregate support a model in which the rate of the loop opening for TIM is dependent on the ligand and results in opening rates in the presence of the product that are comparable to enzymatic throughput, kcat. PMID- 12627962 TI - Physiological concentrations of divalent magnesium ion activate the serine/threonine specific protein kinase ERK2. AB - Extracellular regulated protein kinase 2 (ERK2) is a eukaryotic protein kinase whose activity is regulated by phorbol esters, serum, and growth factors, and displays enhanced activity in several human tumors. Despite its important biological function, its mechanism of catalysis and mode of regulation are poorly understood. Recently, we showed that in the presence of 10 mM magnesium chloride, ERK2 phosphorylates the transcription factor Ets-1 through a random-ordered ternary-complex mechanism [Waas, W. F., and Dalby, K. N. (2002) J. Biol. Chem. 277, 12532]. Now we provide kinetic evidence that ERK2 must bind two divalent magnesium ions to facilitate catalysis at a physiologically relevant rate, because a second magnesium ion promotes both MgATP2- binding and phosphoryl transfer. The velocity dependence on magnesium at saturating concentrations of the protein substrate, Ets138, over a range of ATP4- and Mg2+ ion concentrations, supports the notion that magnesium is an essential activator of ERK2. At high (> or = 1 mM) concentrations of ATP4-, the velocity dependence on total Mg2+ is sigmoidal, but plateaus at high concentrations of free Mg2+, where the enzyme is fully activated. At concentrations of Mg2+ of < or = 4 mM, the velocity dependence on ATP4- displays a peak when the concentration of ATP4- approaches that of total Mg2+ and tends to zero at high concentrations of ATP4-, where the enzyme is predominantly unactivated. The observed velocity dependencies are consistent with the notion that ERK2*Etsdelta138 complexes and ATP4- compete for the same pool of Mg2+ ions in solution. No binding of ATP4- (0-2.5 mM) by ERK2 (65 microM) can be detected using isothermal titration calorimetry at 27 degrees C, pH 8.0, and an ionic strength of 0.15 M (KCl), suggesting that the complex, MgATP2-, is the true substrate for ERK2. In contrast, 5-iodotubericidin binds ERK2 tightly (K(d) = 1 microM) and displays a competitive inhibition pattern toward MgATP2- and a mixed pattern toward free Mg2+, suggesting that the binding of Mg2+ before MgATP2- is not compulsory. PMID- 12627961 TI - Mapping out regions on the surface of the aspartate receptor that are essential for kinase activation. AB - The aspartate receptor of bacterial chemotaxis is representative of a large family of taxis receptors widespread in prokaryotes. The homodimeric receptor associates with cytoplasmic components to form a receptor-kinase signaling complex. Within this complex the receptor is known to directly contact the histidine kinase CheA, the coupling protein CheW, and other receptor dimers. However, the locations and extents of the contact regions on the receptor surface remain ambiguous. The present study applies the protein-interactions-by-cysteine modification (PICM) method to map out surfaces on the aspartate receptor that are essential for kinase stimulation in the assembled receptor-kinase complex. The approach utilizes 52 engineered cysteine positions scattered over the surface of the receptor periplasmic and cytoplasmic domains. When the bulky, anionic probe 5 fluorescein-maleimide is coupled to these positions, large effects on receptor mediated kinase stimulation are observed at eight cytoplasmic locations. By contrast, no large effects are observed for probe attachment at exposed positions in the periplasmic domain. The results indicate that essential receptor surface regions are located near the hairpin turn at the distal end of the cytoplasmic domain and in the cytoplasmic adaptation site region. These surface regions include the docking sites for CheA, CheW, and other receptor dimers, as well as surfaces that transmit information from the receptor adaptation sites to the kinase. Smaller effects observed in the cytoplasmic linker or HAMP region suggest this region may also play a role in kinase regulation. A comparison of the activity perturbations caused by a dianionic, bulky probe (5-fluorescein maleimide), a zwitterionic, bulky probe (5-tetramethyl-rhodamine-maleimide), and a nonionic, smaller probe (N-ethyl-maleimide) reveals the roles of probe size and charge in generating the observed effects on kinase activity. Overall, the results indicate that interactions between the periplasmic domains of different receptor dimers are not required for kinase activation in the signaling complex. By contrast, the observed spatial distribution of protein contact surfaces on the cytoplasmic domain is consistent with both (i) distinct docking sites for cytoplasmic proteins and (ii) interactions between the cytoplasmic domains of different dimers to form a trimer-of-dimers. PMID- 12627963 TI - Structural and mechanistic studies on ThiO, a glycine oxidase essential for thiamin biosynthesis in Bacillus subtilis. AB - The thiO gene of Bacillus subtilis encodes an FAD-dependent glycine oxidase. This enzyme is a homotetramer with a monomer molecular mass of 42 kDa. In this paper, we demonstrate that ThiO is required for the biosynthesis of the thiazole moiety of thiamin pyrophosphate and describe the structure of the enzyme with N acetylglycine bound at the active site. The closest structural relatives of ThiO are sarcosine oxidase and d-amino acid oxidase. The ThiO structure, as well as the observation that N-cyclopropylglycine is a good substrate, supports a hydride transfer mechanism for the enzyme. A mechanistic proposal for the role of ThiO in thiazole biosynthesis is also described. PMID- 12627964 TI - Mechanistic characterization of the HDV genomic ribozyme: classifying the catalytic and structural metal ion sites within a multichannel reaction mechanism. AB - Prior studies of the metal ion dependence of the self-cleavage reaction of the HDV genomic ribozyme led to a mechanistic framework in which the ribozyme can self-cleave by multiple Mg2+ ion-independent and -dependent channels [Nakano et al. (2001) Biochemistry 40, 12022]. In particular, channel 2 involves cleavage in the presence of a structural Mg2+ ion without participation of a catalytic divalent metal ion, while channel 3 involves both structural and catalytic Mg2+ ions. In the present study, experiments were performed to probe the nature of the various divalent ion sites and any specificity for Mg2+. A series of alkaline earth metal ions was tested for the ability to catalyze self-cleavage of the ribozyme under conditions that favor either channel 2 or channel 3. Under conditions that populate primarily channel 3, nearly identical K(d)s were obtained for Mg2+, Ca2+, Ba2+, and Sr2+, with a slight discrimination against Ca2+. In contrast, under conditions that populate primarily channel 2, tighter binding was observed as ion size decreases. Moreover, [Co(NH3)6]3+ was found to be a strong competitive inhibitor of Mg2+ for channel 3 but not for channel 2. The thermal unfolding of the cleaved ribozyme was also examined, and two transitions were found. Urea-dependent studies gave m-values that allowed the lower temperature transition to be assigned to tertiary structure unfolding. The effects of high concentrations of Na+ on the melting temperature for RNA unfolding and the reaction rate revealed ion binding to the folded RNA, with significant competition of Na+ (Hill coefficient of 1.5-1.7) for a structural Mg2+ ion and an unusually high intrinsic affinity of the structural ion for the RNA. Taken together, these data support the existence of two different classes of metal ion sites on the ribozyme: a structural site that is inner sphere with a major electrostatic component and a preference for Mg2+, and a weak catalytic site that is outer sphere with little preference for a particular divalent ion. PMID- 12627965 TI - Implication of phosphatidylinositol 3-kinase membrane recruitment in hydrogen peroxide-induced activation of PI3K and Akt. AB - The effect of tyrosine phosphorylation of PI3K on its enzymatic activity is quite controversial, and the molecular mechanism by which ROS trigger PI3K membrane relocation is unclear. Therefore, we investigated the regulatory mechanism of hydrogen peroxide-induced PI3K activation in DT40 cells, utilizing genetic and pharmacological approaches. Our results revealed that hydrogen peroxide induced tyrosine phosphorylation of the p110 but not the p85 subunit of PI3K in DT40 cells. This phosphorylation was intact in Btk- and Cbl-deficient DT40 cells, but was drastically suppressed in Lyn, Syk, or BCAP-deficient DT40 cells. Tyrosine phosphorylation of p110 did not alter its catalytic activity, and hydrogen peroxide stimulation did not cause an increase in the intrinsic PI3K activity; however, hydrogen peroxide stimulation did induce PI(3,4,5)P3 accumulation and activate Akt. The activation of Akt, as monitored by its ability to phosphorylate GSK-3alpha/beta and by its S473 phosphorylation, was strictly dependent on PI3K activity. Under our conditions, hydrogen peroxide-induced PI3K and Akt activation was independent of Lyn, Syk, Cbl, BCAP, or Ras when each was eliminated individually either by mutation or by a specific inhibitor. In comparison, Akt activation by B cell receptor cross-linking was dependent on BCAP. In addition, hydrogen peroxide treatment caused an increase in the amount of p85 PI3K associated with the particulate fraction. Together, these results indicate that the hydrogen peroxide-induced PI3K and Akt activation in DT40 cells was achieved through PI3K membrane recruitment to its substrate site, thereby enabling PI3K to maximize its catalytic efficiency. PMID- 12627966 TI - Sequences in the intracellular loops of the yeast pheromone receptor Ste2p required for G protein activation. AB - The alpha-factor receptor of the yeast Saccharomyces cerevisiae encoded by the STE2 gene is a member of the large family of G protein-coupled receptors (GPCRs) that mediate multiple signal transduction pathways. The third intracellular loop of GPCRs has been identified as a likely site of interaction with G proteins. To determine the extent of allowed substitutions within this loop, we subjected a stretch of 21 amino acids (Leu228-Leu248) to intensive random mutagenesis and screened multiply substituted alleles for receptor function. The 91 partially functional mutant alleles that were recovered contained 96 unique amino acid substitutions. Every position in this region can be replaced with at least two other types of amino acids without a significant effect on function. The tolerance for nonconservative substitutions indicates that activation of the G protein by ligand-bound receptors involves multiple intramolecular interactions that do not strongly depend on particular sequence elements. Many of the functional mutant alleles exhibit greater than normal levels of signaling, consistent with an inhibitory role for the third intracellular loop. Removal of increasing numbers of positively charged residues from the loop by site-directed mutagenesis causes a progressive loss of signaling function, indicating that the overall net charge of the loop is important for receptor function. Introduction of negatively charged residues also leads to a reduced level of signaling. The defects in signaling caused by substitution of charged amino acids are not caused by changes in the abundance of receptors at the cell surface. PMID- 12627967 TI - Mutants of phosphorylase a altered in recognition by protein phosphatase-1. AB - To develop our knowledge of specificity determinants for protein phosphatase-1, mutants of phosphorylase b have been converted to phosphorylase a and examined for their efficacy as substrates for protein phosphatase-1. Mutants focused on the N-terminal primary sequence surrounding the phosphoserine (R16A, R16E, and I13G) and at a site that interacts with the phosphoserine in phosphorylase a, (R69K and R69E). The success achieved studying protein kinase substrate specificity with peptide substrates has not extended to protein phosphatases. Protein phosphatases are believed to recognize higher order structure in substrates in addition to the primary sequence surrounding the phosphoserine or threonine. Peptide studies with protein phosphatase-1 have revealed a preference for basic residues N-terminal to the phosphoserine. Arginine 16 in phosphorylase a may be a positive determinant. In this work, protein phosphatase-1 preferred the positive charge on arginine 16. R16A exhibited a similar K(m) but reduced V(max), and R16E had an increased K(m) and a decreased V(max) when compared to phosphorylase. I13G had a similar K(m) but an increased V(max). The R69 mutants were also dephosphorylated preferentially over phosphorylase a. The K(m) for R69K was unchanged but had a higher V(max). R69E exhibited the most changes, with a 4 fold increase in K(m) and a 10-fold increase in V(max). These results suggest that proper presentation of the phosphoserine can greatly affect the rate of dephosphorylation. PMID- 12627968 TI - A mutation in the lactose permease of Escherichia coli that decreases conformational flexibility and increases protein stability. AB - Lactose permease with Cys154 --> Gly (helix V) binds substrate with high affinity but catalyzes little or no transport. The purified, detergent-solubilized mutant protein exhibits much greater thermal stability than the wild type and little tendency to aggregate. Stabilization is also observed in vivo with an unstable mutant that is expressed at significantly higher levels when the Cys154 --> Gly mutation is introduced. In addition, ligand-induced conformational changes are markedly reduced or abolished by the Cys154 --> Gly mutation: (i) Although the fluorescence of purified single Trp33 (helix I) permease is enhanced by ligand binding, introduction of the Cys154 --> Gly mutation abolishes the effect. (ii) The rate of 2-(4'-maleimidylanilino)naphthalene-6-sulfonic acid (MIANS) labeling of permease with a single Cys residue in place of Val331 (helix X) is increased in the presence of ligand but reduced when the Cys154 --> Gly mutation is present. (iii) Fluorescence emission intensity of MIANS-labeled single Cys331 permease is enhanced and blue shifted in the Cys154 --> Gly mutant background, indicating that the latter mutation causes position 331 to become exposed to a less polar environment. The results indicate that the Cys154 --> Gly mutation causes a more compact structure and decreased conformational flexibility, an alteration that specifically blocks the structural changes necessary for substrate translocation with little or no effect on ligand binding. PMID- 12627969 TI - Isolation, characterization and electron microscopic single particle analysis of the NADH:ubiquinone oxidoreductase (complex I) from the hyperthermophilic eubacterium Aquifex aeolicus. AB - The proton-translocating NADH:ubiquinone oxidoreductase (complex I) has been purified from Aquifex aeolicus, a hyperthermophilic eubacterium of known genome sequence. The purified detergent solubilized enzyme is highly active above 50 degrees C. The specific activity for electron transfer from NADH to decylubiquinone is 29 U/mg at 80 degrees C. The A. aeolicus complex I is completely sensitive to rotenone and 2-n-decyl-quinazoline-4-yl-amine. SDS polyacrylamide gel electrophoresis shows that it may contain up to 14 subunits. N terminal amino acid sequencing of the bands indicates the presence of a stable subcomplex, which is composed of subunits E, F, and G. The isolated complex is highly stable and active in a temperature range from 50 to 90 degrees C, with a half-life of about 10 h at 80 degrees C. The activity shows a linear Arrhenius plot at 50-85 degrees C with an activation energy at 31.92 J/mol K. Single particle electron microscopy shows that the A. aeolicus complex I has the typical L-shape. However, visual inspection of averaged images reveals many more details in the external arm of the complex than has been observed for complex I from other sources. In addition, the angle (90 degrees ) between the cytoplasmic peripheral arm and the membrane intrinsic arm of the complex appears to be invariant. PMID- 12627970 TI - Thiol regulation of the thylakoid electron transport chain--a missing link in the regulation of photosynthesis? AB - Avoidance of over-reduction of the chloroplast ferredoxin pool is of paramount importance for plants in avoiding oxidative stress. The redox state of this pool can be controlled through regulation of the thylakoid electron transport chain. A model is presented for regulation of this chain via a thiol reduction mechanism, possibly involving a thioredoxin. It is shown in isolated thylakoids that electron transport is inhibited by the thiol reducing agent dithiothreitol. The kinetics of this reduction are rapid and readily reversible. The midpoint redox potential is -365 mV at pH 7.7, with a pH dependency of about -90 mV/pH. At physiological pH values, this places the potential of the species titrated between that of ferredoxin and NADPH and thus in the right potential range to be regulating the redox poise of the ferredoxin pool. This is also close to the potential of NADPH-malate dehydrogenase, an enzyme known to be regulated by thioredoxin. Regulation of electron transport by thioredoxin provides a mechanistic link between the regulation of photosynthesis and gene expression by sugars and the redox regulation of gene expression mediated through the plastoquinone pool. PMID- 12627971 TI - Near-IR irradiation of the S2 state of the water oxidizing complex of photosystem II at liquid helium temperatures produces the metalloradical intermediate attributed to S1Y(Z*). AB - Near-IR (NIR) excitation at liquid He temperatures of photosystem II (PSII) membranes from the cyanobacterium Synechococcus vulcanus or from spinach poised in the S2 state results in the production of a g = 2.035 EPR resonance, reminiscent of metalloradical signals. The signal is smaller in the spinach preparations, but it is significantly enhanced by the addition of exogenous quinones. Ethanol (2-3%, v/v) eliminates the ability to trap the signal. The g = 2.035 signal is identical to the one recently obtained by Nugent et al. by visible-light illumination of the S1 state, and preferably assigned to S1Y(Z*) [Nugent, J. H. A., Muhiuddin, I. P., and Evans, M. C. W. (2002) Biochemistry 41, 4117-4126]. The production of the g = 2.035 signal by liquid He temperature NIR excitation of the S2 state is paralleled by a significant reduction (typically 40 45% in S. vulcanus) of the S2 state multiline signal. This is in part due to the conversion of the Mn cluster to higher spin states, an effect documented by Boussac et al. [Boussac, A., Un, S., Horner, O., and Rutherford, A. W. (1998) Biochemistry 37, 4001-4007], and in part due to the conversion to the g = 2.035 configuration. Following the decay of the g = 2.035 signal at liquid helium temperatures (decay halftimes in the time range of a few to tens of minutes depending on the preparation), annealing at elevated temperatures (-80 degrees C) results in only partial restoration of the S2 state multiline signal. The full size of the signal can be restored by visible-light illumination at -80 degrees C, implying that during the near-IR excitation and subsequent storage at liquid helium temperatures recombination with Q(A-) (and therefore decay of the S2 state to the S1 state) occurred in a fraction of centers. In support of this conclusion, the g = 2.035 signal remains stable for several hours (at 11 K) in centers poised in the S2...Q(A) configuration before the NIR excitation. The extended stability of the signal under these conditions has allowed the measurement of the microwave power saturation and the temperature dependence in the temperature range of 3.8-11 K. The signal intensity follows Curie law temperature dependence, which suggests that it arises from a ground spin state, or a very low-lying excited spin state. The P1/2 (microwave power at half saturation) value is 1.7 mW at 3.8 K and increases to 96 mW at 11 K. The large width of the g = 2.035 signal and its relatively fast relaxation support the assignment to a radical species in the proximity of the Mn cluster. The whole phenomenology of the g = 2.035 signal production is analogous to the effects of NIR excitation on the S3 state [Ioannidis, N., Nugent, J. H. A., and Petrouleas, V. (2002) Biochemistry 41, 9589-9600] producing an S2'Y(Z*) intermediate. In the present case, the intermediate is assigned to S1Y(Z*). The NIR-induced increase in the oxidative capability of the Mn cluster is discussed in relation to the photochemical properties of a Mn(III) ion that exists in both S2 and S3 states. The EPR properties of the S1Y(Z*) intermediate cannot be reconciled easily with our current understanding of the magnetic properties of the S1 state. It is suggested that oxidation of tyr Z alters the magnetic properties of the Mn cluster via exchange of a proton. PMID- 12627972 TI - The two biosynthetic routes leading to phosphatidylcholine in yeast produce different sets of molecular species. Evidence for lipid remodeling. AB - Phosphatidylcholine (PC), a major lipid class in the membranes of eukaryotes, is synthesized either via the triple methylation of phosphatidylethanolamine (PE) or via the CDP-choline route. To investigate whether the two biosynthetic routes contribute differently to the steady-state profile of PC species, i.e., PC molecules with specific acyl chain compositions, the pools of newly synthesized PC species were monitored by labeling Saccharomyces cerevisiae with deuterated precursors of the two routes, (methyl-D3)-methionine and (D13)-choline, respectively. Electrospray ionization tandem mass spectrometry (ESI-MS/MS) revealed that the two PC biosynthetic pathways yield different sets of PC species, with the CDP-choline route contributing most to the molecular diversity. Moreover, yeast was shown to be capable of remodeling PC by acyl chain exchange at the sn-1 position of the glycerol backbone. Remodeling was found to be required to generate the steady-state species distribution of PC. This is the first study demonstrating a functional difference between the two biosynthetic routes in yeast. PMID- 12627973 TI - Interaction of myosin subfragment 1 with forms of monomeric actin. AB - The ability of myosin subfragment 1 to interact with monomeric actin complexed to sequestering proteins was tested by a number of different techniques such as affinity absorption, chemical cross-linking, fluorescence titration, and competition procedures. For affinity absorption, actin was attached to agarose immobilized DNase I. Both chymotryptic subfragment 1 isoforms (S1A1 and S1A2) were retained by this affinity matrix. Fluorescence titration employing pyrenyl actin in complex with deoxyribonuclease I (DNase I) or thymosin beta4 demonstrated S1 binding to these actin complexes. A K(D) of 5 x 10(-8) M for S1A1 binding to the actin-DNase I complex was determined. Fluorescence titration did not indicate binding of S1 to actin in complex with gelsolin segment 1 (G1) or vitamin D-binding protein (DBP). However, fluorescence competition experiments and analysis of tryptic cleavage patterns of S1 indicated its interaction with actin in complex with DBP or G1. Formation of the ternary DNase I-acto-S1 complex was directly demonstrated by sucrose density sedimentation. S1 binding to G-actin was found to be sensitive to ATP and an increase in ionic strength. Actin fixed in its monomeric state by DNase I was unable to significantly stimulate the Mg2+ dependent S1-ATPase activity. Both wild-type and a mutant of Dictyostelium discoideum myosin II subfragment 1 containing 12 additional lysine residues within an insertion of 20 residues into loop 2 (K12/20-Q532E) were found to also interact with actin-DNase I complex. Binding of the K12/20-Q532E mutant to the actin-DNase I complex occurred with higher affinity than wild-type S1 and was less sensitive to mono- and divalent cations. PMID- 12627974 TI - Tauroursodeoxycholic acid prevents Bax-induced membrane perturbation and cytochrome C release in isolated mitochondria. AB - Bax is a potent pro-apoptotic member of the Bcl-2 protein family that localizes to the mitochondrial membrane during apoptosis. Tauroursodeoxycholic acid (TUDCA) modulates the apoptotic threshold, in part, by preventing Bax translocation both in vitro and in vivo. The mechanisms by which Bax induces and TUDCA inhibits release of cytochrome c are unclear. We show here that recombinant Bax protein induced cytochrome c release in isolated mitochondria without detectable swelling. Co-incubation with TUDCA prevented efflux of mitochondrial factors and proteolytic processing of caspases in cytosolic extracts. Spectroscopic analyses of mitochondria exposed to Bax revealed increased polarity and fluidity of the membrane lipid core as well as altered protein order, indicative of Bax binding, together with loss of spin-label paramagnetism, characteristic of oxidative damage. TUDCA markedly abrogated the Bax-induced membrane perturbation. In conclusion, our results indicate that Bax protein directly induces cytochrome c release from mitochondria through a mechanism that does not require the permeability transition. Rather, it is accompanied by changes in the organization of membrane lipids and proteins. TUDCA is a potent inhibitor of Bax association with mitochondria. Thus, TUDCA modulates apoptosis by suppressing mitochondrial membrane perturbation through pathways that are also independent of the mitochondrial permeability transition. PMID- 12627975 TI - Structure-function analysis of a series of novel GIP analogues containing different helical length linkers. AB - Glucose-dependent insulinotropic polypeptide (GIP1-42) is a potent glucose lowering intestinal peptide hormone. The equipotent GIP1-30NH2 was structurally modified by linking N- and C-terminal fragments with several different linkers. Substitution of the middle region of GIP by a flexible aminohexanoic linker resulted in greatly reduced binding affinity and reduction or complete loss of bioactivity. Connection of the bioactive domains GIP1-14 and GIP19-30NH2 by EKEK or AAAA linkers resulted in peptide agonists with approximately 3-4-fold increased bioactivity as compared to GIP1-30NH2. Conformational analysis by CD spectroscopy of GIP fragments and analogues suggests a helical region in the C terminal (19-30) portion of GIP. It was demonstrated that stabilization of this C terminal helical region by the introduction of helical linkers favored binding and activation of the GIP receptor. Our results suggest an important contribution of a direct interaction of the first 14 amino acids with the GIP receptor, an appropriate relative orientation of N- and C-terminal parts of GIP, and the presence of helical linkers to be essential for bioactivity. PMID- 12627976 TI - Arabidopsis thaliana Ogg1 protein excises 8-hydroxyguanine and 2,6-diamino-4 hydroxy-5-formamidopyrimidine from oxidatively damaged DNA containing multiple lesions. AB - A functional homologue of eukaryotic Ogg1 proteins in the model plant Arabidopsis thalianahas recently been cloned, isolated, and characterized [Garcia-Ortiz, M. V., Ariza, R. R., and Roldan-Arjona, T. (2001) Plant Mol. Biol. 47, 795-804]. This enzyme (AtOgg1) exhibits a high degree of sequence similarity in several highly conserved regions with Saccharomyces cerevisiae, Drosophila melanogaster, and human Ogg1 proteins. We investigated the substrate specificity and kinetics of AtOgg1 for excision of modified bases from oxidatively damaged DNA that contained multiple pyrimidine- and purine-derived lesions. Two different DNA substrates prepared by exposure to ionizing radiation in aqueous solution under N2O or air were used for this purpose. Gas chromatography/isotope-dilution mass spectrometry was applied to identify and quantify modified bases in DNA samples. Of the 17 modified bases identified in DNA samples, only 8-hydroxyguanine and 2,6 diamino-4-hydroxy-5-formamidopyrimidine were significantly excised from both DNA substrates. This is in agreement with the substrate specificities of other eukaryotic Ogg1 proteins that had previously been studied under identical conditions. Excision depended on incubation time, enzyme concentration, and substrate concentration and followed Michaelis-Menten kinetics. A significant dependence of excision on the nature of DNA substrate was observed in accord with previous studies on other DNA glycosylases. A comparison of excision kinetics pointed to significant differences between AtOgg1 and other Ogg1 proteins. We also investigated the effect of base-pairing on the excision using double stranded oligodeoxynucleotides that contained 8-OH-Gua paired with each of the four DNA bases. The activity of AtOgg1 was most effective on the 8-OH-Gua:C pair with some or very low activity on other pairs in agreement with the activity of other Ogg1 proteins. The results unequivocally show that AtOgg1 possesses common substrates with other eukaryotic Ogg1 proteins albeit significant differences between their excision kinetics. PMID- 12627977 TI - HU binding to bent DNA: a fluorescence resonance energy transfer and anisotropy study. AB - HU, an architectural DNA-binding protein, either stabilizes DNA in a bent conformation or induces a bend upon binding to give other proteins access to the DNA. In this study, HU binding affinity for a bent DNA sequence relative to a linear sequence was investigated using fluorescence anisotropy measurements. A static bend was achieved by the introduction of two phased A4T4 tracts in a 20 bp duplex. Binding affinity for 20 bp duplexes containing two phased A-tracts in either a 5'-3' or 3'-5' orientation was found to be almost 10-fold higher than HU binding to a random sequence 20 bp duplex (6.1 vs 0.68 microM(-1)). The fluorescence technique of resonance energy transfer was used to quantitatively determine the static bend of the DNA duplexes and the HU-induced bend. DNA molecules were 5'-end labeled with fluorescein as the donor or rhodamine as the acceptor. From the efficiency of energy transfer, the end-to-end distance of the DNA duplexes was calculated. The end-to-end distance relative to DNA contour length (R/R(C)) yields a bend angle for the A-tract duplex of 45 +/- 7 degrees in the absence of HU and 70 +/- 3 degrees in the presence of HU. The bend angle calculated for the T4A4 tract duplex was 62 +/- 4 degrees after binding two HU dimers. Fluorescence anisotropy measurements reveal that HU binds in a 1:1 stoichiometry to the A4T4 tract duplex but a 2:1 stoichiometry to the T4A4 tract and random sequence duplex. These findings suggest that HU binding and recognition of DNA may be governed by a structural mechanism. PMID- 12627978 TI - Demonstration of two independently folding domains in the alpha subunit of bacterial luciferase by preferential ligand binding-induced stabilization. AB - The alpha subunit of bacterial luciferase unfolds and refolds reversibly by a three-state mechanism in urea-containing buffer. It has been proposed that the three-state unfolding of the alpha subunit arises from a stepwise unfolding of a C-terminal folding domain at lower concentrations of urea, followed by unfolding of the N-terminal domain at higher concentrations of urea (Noland, B. W., Dangott, L. J., and Baldwin, T. O. (1999) Biochemistry 38, 16136-16145). The location of an anion binding site in the proposed N-terminal folding domain allowed the folding mechanism to be probed in the context of the intact polypeptide. Anions preferentially stabilized the N-terminal domain in a concentration-dependent manner. The polyvalent anions sulfate and phosphate were found to be more stabilizing than monovalent chloride ion. Cations did not show a similar stabilizing effect, demonstrating that the stabilization was due to the anions alone. The purified N-terminal domain prepared by limited proteolysis and anion exchange chromatography was found to refold cooperatively with a midpoint approximately that of the second unfolding transition of the alpha subunit. Phosphate ion stabilized this fragment to roughly the same extent as it did the alpha subunit. The results presented are consistent with the proposed two-domain folding model and demonstrate that anion binding to the N-terminal folding domain stabilizes the alpha subunit of bacterial luciferase. PMID- 12627979 TI - Random-order ternary complex reaction mechanism of serine acetyltransferase from Escherichia coli. AB - Although serine acetyltransferase (SAT) from Escherichia coli is homologous with a number of bacterial enzymes that catalyze O-acetyl transfer by a sequential (ternary complex) mechanism, it has been suggested, from experiments with the nearly identical enzyme from Salmonella typhimurium, that the reaction could proceed via an acetyl-enzyme intermediate. To resolve the matter, the E. coli gene for SAT was overexpressed and the enzyme purified 13-fold to homogeneity. The results of a steady-state kinetic analysis of the forward reaction are diagnostic for a ternary complex mechanism, and the response of SAT to dead-end inhibitors indicates a random order for the addition of substrates. The linearity of primary double-reciprocal plots, in the presence and absence of dead-end inhibitors, argues that interconversion of ternary complexes is not significantly faster than kcat, whereas substrate inhibition by serine suggests that breakdown of the SAT.CoA binary complex is rate-determining. The results of equilibrium isotope exchange experiments, for both half-reactions, rule out a "ping-pong" mechanism involving an acetyl-enzyme intermediate, and a pre-steady-state kinetic analysis of the turnover of AcCoA supports such a conclusion. Kinetic data for the reverse reaction (acetylation of CoA by O-acetylserine) are also consistent with a steady-state random-order mechanism, wherein both the breakdown of the SAT*serine complex and the interconversion of ternary complexes are partially rate-determining. PMID- 12627980 TI - Aluminum exchange between citrate and human serum transferrin and interaction with transferrin receptor 1. AB - The kinetics and thermodynamics of Al(III) exchange between aluminum citrate (AlL) and human serum transferrin were investigated in the 7.2-8.9 pH range. The C-site of human serum apotransferrin in interaction with bicarbonate removes Al(III) from Al citrate with an exchange equilibrium constant K1 = (2.0 +/- 0.6) x 10(-2); a direct second-order rate constant k1 = 45 +/- 3 M(-1) x s(-1); and a reverse second-order rate constant k(-1) = (2.3 +/- 0.5) x 10(3) M(-1) x s(-1). The newly formed aluminum-protein complex loses a single proton with proton dissociation constant K1a = (15 +/- 3) nM to yield a first kinetic intermediate. This intermediate then undergoes a modification in its conformation followed by two proton losses; first-order rate constant k2 = (4.20 +/- 0.02) x 10(-2) s(-1) to produce a second kinetic intermediate, which in turn undergoes a last slow modification in the conformation to yield the aluminum-loaded transferrin in its final state. This last process rate-controls Al(III) uptake by the N-site of the protein and is independent of the experimental parameters with a constant reciprocal relaxation time tau3(-1) = (6 +/- 1) x 10(-5) x s(-1). The affinities involved in aluminum uptake by serum transferrins are about 10 orders of magnitude lower than those involved in the uptake of iron. The interactions of iron-loaded transferrins with transferrin receptor 1 occur with average dissociation constants of 3 +/- 1 and 5 +/- 1 nM for the only C-site iron-loaded and of 6.0 +/- 0.6 and 7 +/- 0.5 nM for the iron-saturated ST in the absence or presence of CHAPS, respectively. No interaction is detected between receptor 1 and aluminum-saturated or mixed C-site iron-loaded/N-site aluminum-loaded transferrin under the same conditions. The fact that aluminum can be solubilized by serum transferrin in biological fluids does not necessarily imply that its transfer from the blood stream to cytoplasm follows the receptor-mediated pathway of iron transport by transferrins. PMID- 12627981 TI - Membrane composition affects the reversibility of annexin A2t binding to solid supported membranes: a QCM study. AB - By means of the quartz crystal microbalance (QCM) technique, we investigated the interaction of porcine heterotetrametric annexin A2t with solid supported lipid membranes. Dissociation and rate constants of annexin A2t binding to various lipid mixtures were determined as a function of Ca2+ concentrations in solution. In contrast to what has been observed for annexin A1, the binding affinity and kinetics of annexin A2t binding are not influenced by cholesterol. In the experimental setup chosen, the annexin A2t binding is strictly Ca2+-dependent and only affected by the amount of phosphatidylserine (PS) in the membrane and the Ca2+ concentration in solution. By Ca2+-titration experiments at constant annexin A2t concentration, we investigated the reversibility of annexin A2t adsorption and desorption. Surprisingly, Ca2+-titration curves display a significant hysteresis. Protein desorption curves starting from annexin A2t bound to the membrane at 1 mM CaCl2 exhibit high cooperativity with half-maximum Ca2+ concentrations in the submicromolar range. However, protein adsorption curves starting from an EGTA-containing solution with soluble annexin A2t always show two inflection points upon addition of Ca2+ ions. These two inflection points may be indicative of two protein populations differently bound to the solid-supported membrane. The ratio of these two annexin A2t populations depends on the amount of PS molecules and cholesterol in the membrane as well as on the Ca2+ concentration. We propose a model discussing the results obtained in terms of two binding sites differing in their affinity due to lipid rearrangement. PMID- 12627982 TI - Multiple pathways for mineral core formation in mammalian apoferritin. The role of hydrogen peroxide. AB - Human ferritins sequester and store iron as a stable FeOOH((s)) mineral core within a protein shell assembled from 24 subunits of two types, H and L. Core mineralization in recombinant H- and L-subunit homopolymer and heteropolymer ferritins and several site-directed H-subunit variants was investigated to determine the iron oxidation/hydrolysis chemistry as a function of iron flux into the protein. Stopped-flow absorption spectrometry, UV spectrometry, and electrode oximetry revealed that the mineral core forms by at least three pathways, not two as previously thought. They correspond to the ferroxidase, mineral surface, and the Fe(II) + H2O2 detoxification reactions, respectively: [see reactions]. The H subunit catalyzed ferroxidase reaction 1 occurs at all levels of iron loading of the protein but decreases with increasing iron added (48-800 Fe(II)/protein). Reaction 2 is the dominant reaction at 800 Fe(II)/protein, whereas reaction 3 occurs largely at intermediate iron loadings of 100-500 Fe(II)/protein. Some of the H2O2 produced in reaction 1 is consumed in the detoxification reaction 3; the 2/1 Fe(II)/H2O2 stoichiometry of reaction 3 minimizes hydroxyl radical production during mineralization. Human L-chain ferritin and H-chain variants lacking functional nucleation and/or ferroxidase sites deposit their iron largely through the mineral surface reaction 2. H2O2 is shown to be an intermediate product of dioxygen reduction in L-chain as well as in H-chain and H-chain variant ferritins. PMID- 12627983 TI - A simulation model for estimating direct costs of type 1 diabetes prevention. AB - BACKGROUND: The ongoing Type 1 Diabetes Prediction and Prevention Project in Finland (DIPP) is based on screening of genetic type 1 diabetes mellitus susceptibility, subsequent autoantibody follow-up and experimental preventive treatment with nasal insulin. OBJECTIVE: To analyse direct costs of type 1 diabetes prevention therapy with nasal insulin as it is now being studied in the DIPP project, and as it might be used as a part of routine healthcare in Finland. DATA AND METHODS: For the purposes of cost analysis, two different diabetes prevention models were constructed. The research-oriented model followed accurately the DIPP protocol and the practice-oriented model was based on the estimates of a panel of experts on how the prevention would be conducted as a part of the routine healthcare in Finland. To take into account the uncertainty and variability attached to the use of resources, a Monte Carlo simulation model was utilised. The costs of the two models comprising 500 iterations each were simulated using the Monte Carlo model. STUDY PERSPECTIVE: This study was performed from the healthcare provider's viewpoint. RESULTS: The total direct costs per person of the research-oriented model were 2102 and 1676 euros (EUR) in the first and second year and those of the practice-oriented model EUR827 and EUR675, respectively (EUR1 approximately dollars US1.1; 2002 values). Subsequently, the costs rose only as a result of the increased use of insulin as the children grew older. After the 15th year, when the age structure of the population in the study had stabilised, the annual direct costs per person were EUR1798 (research-oriented model) and EUR797 (practice-oriented model). CONCLUSIONS: The costs of prevention with nasal insulin are low when compared with estimates of the annual healthcare costs of type 1 diabetes. This study suggests, with some critical assumptions (in particular, that nasal insulin is effective in the prevention of type I diabetes), that a 2 to 3-year delay in the disease onset may make prevention according to the practice-oriented model cost saving. PMID- 12627984 TI - Cost effectiveness of alendronate (fosamax) for the treatment of osteoporosis and prevention of fractures. AB - BACKGROUND: The Fracture Intervention Trial (FIT) demonstrated that the bisphosphonate alendronate reduces the risk of hip, spine and wrist fracture in osteoporotic women by approximately one half. OBJECTIVE: To use data from FIT to conduct a cost-effectiveness analysis of alendronate. DESIGN: A Markov model was developed for a cohort of Swedish women, comparable in relative fracture risk to the women enrolled in the FIT vertebral fracture arm (i.e. age 71 years with low bone mass plus at least one prior spine fracture). The women in the model (with low bone mass and a previous spine fracture) were exposed to alendronate therapy and transitioned over time from a 'well' health state to health states of 'hip fracture', 'spine fracture', 'wrist fracture' or 'death'. All costs were calculated in 2000 Swedish kronors (SEK). TIME HORIZON: In the Markov model our base-case treatment duration was 5 years followed by a 5-year period where the benefit declined linearly to 0. RESULTS: We found that treating 71-year-old osteoporotic women with a prior spine fracture with alendronate resulted in a cost per quality-adjusted life-year (QALY) gained of SEK76000, which is well below the threshold for cost effectiveness of SEK300000. For women aged 65 years, the cost-effectiveness ratio increased to SEK173000 and for women aged 77 years, the cost-effectiveness ratio decreased to SEK52000. CONCLUSIONS: Treating older osteoporotic women with alendronate was more cost effective than treating younger women with osteoporosis, and treating osteoporotic women with prior spine fracture was more cost effective than treating osteoporotic women without prior spine fracture. However, the costs per QALY gained for all populations studied were below generally accepted thresholds for cost effectiveness. PMID- 12627985 TI - The cost effectiveness of two new antiepileptic therapies in the absence of direct comparative data: a first approximation. AB - BACKGROUND: A number of new antiepileptic agents have been introduced within a short period of time. Direct comparisons are not available, and information about the balance between costs and effects for these new therapies is lacking. OBJECTIVE: To introduce a first approximation of the cost effectiveness of the new therapeutic agents (topiramate and lamotrigine) for epilepsy that have been assessed in clinical trials against placebo. METHODS: Without head to head comparative data no formal methods are available to assess the relative cost effectiveness of two products; therefore, a Bayesian approach was developed. The approach starts with the 'proportionality assumption' saying that the differences in healthcare expenditure (less the direct cost of therapy) are directly proportional to the differences in effectiveness. Given this assumption, a therapy that is x times as expensive as an alternative therapy has an equivalent cost-effectiveness profile if the acquisition cost is x times as high. Moreover, simple formulas can be derived to calculate the probabilities that a therapy is dominant (more effective and less expensive) and that it is weakly dominant (more effective and a better cost-effectiveness profile). The approach is applied to data from published fixed dosage, parallel-design studies comparing both topiramate and lamotrigine with placebo. RESULTS: Assuming that the 'proportionality assumption' holds for the medical treatment of epilepsy, and disregarding uncertainties, it is estimated that topiramate may be priced more than 2.2 times its current acquisition cost and still be more cost effective than lamotrigine. Taking uncertainties into account, it is estimated that lamotrigine 500 mg/day is dominated by topiramate 200 mg/day with a probability of 0.875 and by topiramate 400 mg/day with a probability of 0.986. CONCLUSIONS: A simple method can be applied to assess the relative cost effectiveness of two therapies in the absence of direct comparative data. Applying this method to compare topiramate and lamotrigine leads to a strong preference for topiramate. However, to be able to draw this conclusion, some heroic assumptions need to be made. As such the method as developed here only reflects a first approximation. It needs to be used with care and is not intended to replace good comparative research. PMID- 12627986 TI - Resource utilisation and cost analysis of memantine in patients with moderate to severe Alzheimer's disease. AB - BACKGROUND: Alzheimer's disease (AD) is a devastating illness that causes enormous emotional stress to affected families and is associated with substantial medical and nonmedical costs. OBJECTIVE: To determine the effects of 28 weeks of memantine treatment for patients with AD on resource utilisation and costs. STUDY DESIGN AND METHODS: Multicentre, prospective, double-blind, randomised, placebo controlled clinical trial performed in the US. The Wilcoxon-Mann-Whitney test was used to examine the resource utilisation variables and logistic regression models were used for multivariate resource utilisation analyses. Analysis of covariance (ANCOVA) models (log and non-log) were computed to examine costs from a societal perspective. All costs were calculated in 1999 US dollars. STUDY POPULATION: Outpatients with moderate to severe AD. Overall, 252 patients received randomised treatment, and 166 patients (placebo n = 76, memantine n = 90) formed the treated per-protocol (TPP) subset for the health economic analyses, on which the main cost analysis was based. MAIN OUTCOME MEASURE: Resource Utilisation in Dementia (RUD) scale, measuring patient and caregiver resource utilisation, and various sources for cost calculations. RESULTS: Controlling for baseline differences between the groups, significantly less caregiver time was needed for patients receiving memantine than for those receiving placebo (difference 51.5 hours per month; 95% CI -95.27, -7.17; p = 0.02). Analysis of residential status also favoured memantine: time to institutionalisation (p = 0.052) and institutionalisation at week 28 (p = 0.04 with the chi-square test). Total costs from a societal perspective were lower in the memantine group (difference dollars US 1089.74/month [non-overlapping 95% CI for treatment difference -1954.90, 224.58]; p = 0.01). The main differences between the groups were total caregiver costs (dollars US-823.77/month; p = 0.03) and direct nonmedical costs (dollars US 430.84/month; p = 0.07) favouring memantine treatment. Patient direct medical costs were higher in the memantine group (p < 0.01), mainly due to the cost of memantine. CONCLUSION: Resource utilisation and total health costs were lower in the memantine group than the placebo group. The results suggest that memantine treatment of patients with moderate to severe AD is cost saving from a societal perspective. PMID- 12627988 TI - Cost effectiveness of nasal budesonide versus surgical treatment for nasal polyps. AB - OBJECTIVE: To conduct a cost-effectiveness study of nasal budesonide versus surgical treatment in the management of nasal polyps. DESIGN AND METHODS: A decision-tree model reflecting two different treatment strategies for nasal polyps in Sweden was developed. The first strategy was initial polypectomy, performed under three different sets of circumstances: inpatient functional endoscopic surgery, outpatient evulsion with sedation, or outpatient evulsion with local anaesthesia; all treatments were followed by intranasal treatment with budesonide (Rhinocort) 128 microg twice daily. The second strategy was initial intranasal treatment with budesonide 128 microg twice daily. PERSPECTIVE: Healthcare provider perspective. OUTCOME MEASURES AND RESULTS: After 1 month, treatment with nasal budesonide was classified as a success (82.5%) or a failure (17.5%) based on clinical study data. In cases of success, the treatment was continued, and in cases of failure, polypectomy was undertaken, followed by budesonide 128 microg twice daily. Treatments were evaluated after 4 months using prices from the Central Hospital in Skovde, Sweden. The expected reduction in cost from using initial nasal budesonide treatment compared with the different alternatives of polypectomy were 9760 Swedish kronors (SEK) for inpatient functional endoscopic surgery, SEK2747 for outpatient evulsion with sedation, and SEK672 for outpatient evulsion with anaesthesia (1998 values). Nasal budesonide 128 microg twice daily treatment for nasal polyps revealed a potential reduction in costs of 53% compared with the primary surgery approach. CONCLUSION: Initial treatment of nasal polyps with nasal budesonide provides lower costs than treatment with initial polypectomy with maintained effectiveness. PMID- 12627987 TI - Peginterferon alpha-2a (40kD) [Pegasys] improves HR-QOL outcomes compared with unmodified interferon alpha-2a [Roferon-A]: in patients with chronic hepatitis C. AB - BACKGROUND: Use of unmodified interferon alpha-2a in chronic hepatitis C is associated with impaired health-related quality of life during therapy. Treatment with peginterferon alpha-2a (40kD) provides an improved sustained response over unmodified interferon alpha-2a. OBJECTIVES: To compare health-related quality of life during treatment for patients receiving peginterferon alpha-2a (40kD) [Pegasys] versus unmodified interferon alpha-2a [Roferon]. DESIGN: A randomised, international, multicentre, open-label, parallel group study. SETTING: 36 centres worldwide. PATIENTS: Interferon-naive patients (n = 531) with chronic hepatitis C. INTERVENTIONS: Peginterferon alpha-2a (40kD) 180 mirog once a week (n = 267) for 48 weeks or unmodified interferon alpha-2a 6 million IU three times a week for 12 weeks followed by 36 weeks of 3 million IU three times a week (n = 264). MEASUREMENTS: Fatigue Severity Scale and 36-item Short-Form Health Survey (SF 36). RESULTS: At weeks 2 and 12, differences favouring peginterferon alpha-2a (40kD) were seen on seven of eight domains and both summary scores of the SF-36 (p < 0.05 to p < 0.01). At weeks 2, 12 and 24, patients receiving peginterferon alpha-2a (40kD) had less disabling fatigue (p < 0.01) than those receiving unmodified interferon alpha-2a. CONCLUSION: Treatment with peginterferon alpha-2a (40kD) is associated with less disabling fatigue and less impairment in patient functioning and well-being during treatment than unmodified interferon alpha-2a. In addition to safety and efficacy, the impact on health-related quality of life may be an important consideration for physicians when selecting an optimal treatment regimen. PMID- 12627990 TI - Photodynamic therapy: is it a valuable treatment option for actinic keratoses? AB - Photodynamic therapy (PDT) is proposed as an effective therapy with a good safety profile for patients with actinic keratoses. PDT involves the application of a photosensitizer to dysplastic or neoplastic tissue such that when exposed to light of an appropriate wavelength, the target tissue undergoes a cytotoxic reaction. Studies to date have demonstrated that PDT for the treatment of patients with actinic keratoses achieves clearance of lesions with minimal morbidity, maintenance of functional integrity of underlying tissues, and excellent cosmetic results. We present a review of the treatment of patients with actinic keratoses and the role of PDT in this context. PMID- 12627989 TI - Factors affecting UK primary-care costs of managing patients with asthma over 5 years. AB - OBJECTIVE: To determine the effect of age, disease severity and compliance on the annual primary-care cost of managing patients with asthma initially on British Thoracic Society British Guidelines on Asthma Management (BGAM) treatment steps 2/3 over 5 years. DESIGN AND SETTING: A modelling study performed from the perspective of the UK's National Health Service (NHS). STUDY PARTICIPANTS AND INTERVENTIONS: A data set was created comprising 4519 patients with asthma in the DIN-link database who were prescribed twice-daily inhaled corticosteroids and who were on steps 2/3 between 1 January and 31 December 1993. These patients were followed over 5 years. METHODS: Asthma-related primary-care resource utilisation data obtained from the DIN-link database were stratified by patients' age, compliance and BGAM treatment step. Unit costs at 1999-2000 prices were applied to the resource use estimates to determine the mean annual cost per patient. MAIN OUTCOME MEASURES AND RESULTS: High compliance with inhaled corticosteroids was not associated with a reduction in use of other primary-care resources, although the ratio of the number of prescriptions for inhaled corticosteroids to that for short-acting beta(2)-agonists increased, suggesting that patients' asthma was better controlled. Overall, the primary-care cost of managing a patient starting on steps 2/3 was found to be most strongly influenced by whether that patient moved onto steps 4/5 or continued to be managed on the same treatment step. If a patient continued to be managed at steps 2/3, costs were influenced in descending order of impact by compliance, previous BGAM step and the patient's age. CONCLUSIONS: Better compliance with inhaled corticosteroids is likely to lead to better asthma control and fewer asthma attacks. Notwithstanding this, increasing compliance is likely to increase primary-care costs. Consequently increasing healthcare expenditure may be the inevitable consequence of improving asthma control. PMID- 12627991 TI - Erysipelas: recognition and management. AB - Erysipelas is an acute bacterial infection of the dermis and hypodermis that is associated with clinical inflammation. It is a specific clinical type of cellulitis and, as such, it should be studied as a specific entity. Erysipelas is generally caused by group A streptococci; it is highly probable that streptococcal toxins also play a role, which could, in part, help explain the clinical inflammation. Erysipelas of the leg is the main clinical type encountered. The face, arm, and upper thigh are the other most common sites for the occurrence of erysipelas. After a sudden onset, areas of erythema and edema characteristically enlarge with well-defined margins. Athlete's foot is the most common portal of entry for the disease. Erysipelas is generally associated with high fever, and adenopathy and lymphangitis are sometimes present. At the time of diagnosis, it is important to look for clinical markers of severity (local signs and symptoms, general signs and symptoms, co-morbidity, social context) which would necessitate hospitalization. There are many differential diagnoses, particularly in the case of atypical dermo-hypodermitis. Some bacterial infections may have specific clinical aspects or may lead to a diagnosis of cellulitis. Necrotizing cellulitis or fasciitis are life-threatening diseases and a rapid diagnosis is important. Other noninfectious types of cellulitis have been reported in many diseases, both localized or generalized. The biology of typical erysipelas is of little value in diagnosis and a laboratory workup is usually not required. There are few local complications associated with erysipelas; abscess can occur in some patients and septicemia is rare. Recurrence is the more distressing complication. Treatment of patients with erysipelas has been evaluated in a small number of studies. In most of them, erysipelas has been included in therapeutic studies of 'severe cutaneous infections'. This is not justified as in fact erysipelas is usually sensitive to penicillin G. Amoxicillin and macrolides are also effective. However, comparative, cost-analysis studies need to be performed to determine the best therapeutic option. Bed rest with the leg elevated is also important. Anticoagulants are indicated in patients at risk of venous thromboembolism. The portal of entry will also require treatment. Long term antibacterial therapy is required for patients with recurrence. PMID- 12627993 TI - HIV-associated pruritus: etiology and management. AB - With the advent of highly active antiretroviral therapy (HAART), life-threatening opportunistic infection has become less common in patients with HIV infection and longevity has increased dramatically. With increased longevity, the problems of living with a chronic disease have become more prominent in this patient population. Disorders such as fat redistribution and metabolic abnormalities can result from antiviral medications and from HIV disease itself. Pruritus is one of the most common symptoms encountered in patients with HIV. The spectrum of skin diseases in such patients encompasses dermatoses of diverse etiologies; a few are peculiar to patients with HIV while others are not. Some of these conditions may cause severe and sometimes intractable pruritus that provokes scratching, picking, disfigurement, sleep loss, and significant psychological stress. Moreover, the expense of ongoing medical treatments can be daunting. Skin rash can sometimes be the initial presentation of HIV infection or serve as a harbinger of disease progression. Causes of pruritus include skin infections, infestations, papulosquamous disorders, photodermatitis, xerosis, drug reactions, and occasionally lymphoproliferative disorders. Drug eruptions are particularly common in patients who are HIV positive, presumably as a result of immune dysregulation, altered drug metabolism, and polypharmacy. Itching can also result from systemic diseases such as chronic renal failure, liver disease, or systemic lymphoma. Workup of pruritus should include a careful examination of the skin, hair, nails, and mucous membranes to establish a primary dermatologic diagnosis. If no dermatologic cause is found, a systemic cause or medication-related etiology should be sought. Idiopathic HIV pruritus is a diagnosis of exclusion and should only be considered when a specific diagnosis cannot be established. The management of HIV-associated pruritus should be directed at the underlying condition. Phototherapy has been found to be useful in the treatment of several HIV-associated dermatoses and idiopathic pruritus as well. Unfortunately, some of the treatments that have been suggested for patients with HIV are anecdotal or based on small uncontrolled studies. The last decade has seen a surge in the utilization of HAART which, to some degree, reconstitutes the immune system and ameliorates some dermatologic diseases. On the other hand, some skin diseases flare temporarily when HAART is started. Unless frank drug allergy is suspected, HAART does not need to be stopped. PMID- 12627994 TI - Fabry disease: recognition and management of cutaneous manifestations. AB - Fabry disease (angiokeratoma corporis diffusum universale) is a rare, X chromosome-linked lysosomal storage disease. The deficient enzyme, alpha galactosidase A (alpha-gal A), is responsible for the accumulation of neutral glycosphingolipids within vascular endothelial lysosomes of various organs, including skin, kidneys, heart, and brain. The disease manifests primarily in affected hemizygous men and to some extent in heterozygous women ('carriers'). The diagnosis of Fabry disease is made in hemizygous males after the detection of the presence of angiokeratomas, irregularities in sweating, edema, scant body hair, painful sensations, and of cardiovascular, gastrointestinal, renal, ophthalmologic, phlebologic, and respiratory involvement. A deficiency of alpha gal A in serum, leukocytes, tears, tissue specimens, or cultured skin fibroblasts further supports the diagnosis in male patients. Since heterozygous women show angiokeratomas in only about 30% of cases and may have alpha-gal A levels within normal range, genetic analysis is recommended. Current treatment of angiokeratomas of Fabry disease is based mainly on the use of laser systems, including variable pulse width 532nm Neodymium:Yttrium-Aluminum-Garnet (Nd:YAG) laser, 578nm copper vapor laser, and flashlamp-pumped dye laser. When cutaneous and mucous glands are affected, restrictions may be required with regard to the time spent in a warm climate and the amount time spent working or on sporting activities, and may necessitate the use of topical and systemic antiperspirant agents, and topical application of artificial lacrimal fluid and saliva, respectively. For the future, new treatment modalities, including enzyme replacement therapy, substrate deprivation strategies, and gene therapy offer extraordinary options for the cutaneous and visceral lesions in patients with Fabry disease. PMID- 12627992 TI - Staphylococcal scalded skin syndrome: diagnosis and management. AB - Staphylococcal scalded skin syndrome (SSSS) is a common disorder that is usually seen in infants and children and rarely seen in adults. SSSS usually presents with a prodrome of sore throat or conjunctivitis. Extremely tender flaccid bullae, which are Nikolsky sign-positive, develop within 48 hours and commonly affect the flexures; occasionally, large areas of the skin may be involved. The bullae enlarge and rupture easily to reveal a moist erythematous base, which gives rise to the scalded appearance. SSSS in adults is a rare disorder, though there are now over 50 documented cases. Usually SSSS occurs in predisposed individuals, but not all adults have an underlying illness. Whereas mortality in childhood SSSS is approximately 4%, the mortality rate in adults is reported to be greater than 60%. SSSS is caused by an infection with a particular strain of Staphylococcus aureus, which leads to blistering of the upper layer of the skin, by the release of a circulating exotoxin. It has recently been demonstrated that the exfoliative exotoxin responsible for SSSS leads to the cleavage of desmoglein 1 complex, an important desmosomal protein. The same toxins that are responsible for causing SSSS also cause bullous impetigo. There appears to be a relationship between the disease extent, the amount of toxin produced and whether the toxin is released locally or systemically. As a result there is likely to be a spectrum of disease and there are likely to be a number of milder cases of adult SSSS that go undiagnosed. Social improvements and hygiene have led to a dramatic fall in the number of cases of SSSS. Treatment is usually straightforward, when there is no coexistent morbidity and the presentation is mild, but can be demanding if the patient is particularly ill. SSSS is still associated with mortality, particularly when it occurs in adults. PMID- 12627995 TI - Optimizing treatment with topical tazarotene. AB - Tazarotene is a receptor-selective retinoid, which is efficacious in the treatment of patients with psoriasis, acne vulgaris, and photoaging. It normalizes keratinocyte differentiation, reverses keratinocyte hyperproliferation, and has anti-inflammatory effects. Clinical studies have shown that tazarotene 0.1% gel has greater comedolytic activity than tretinoin (Retin-A 0.025% gel, Retin-A Micro 0.1%) and adapalene (Differin) 0.1% gel. Although it is efficacious as monotherapy, tazarotene is more commonly used as part of combination therapy with a topical antibacterial in patients with acne vulgaris, and with a mid- or high-potency topical corticosteroid or with phototherapy in patients with psoriasis. Combination therapy enhances efficacy and tolerability. Tazarotene 0.1% gel, used in combination with mometasone furoate 0.1% cream, was shown in psoriasis clinical trials to be more efficacious than calcipotriene (calcipotriol) ointment used twice daily, or mometasone furoate 0.1% cream used twice daily. Use of tazarotene in conjunction with broad band UVB, narrow band UVB or bath psoralens + UVA (PUVA) results in greater efficacy than with phototherapy alone. Tazarotene should not be administered during pregnancy or in women who are not practicing adequate contraception. Adverse events consist primarily of irritation, peeling, erythema, dryness, burning, and itching. They are most common during the first 1-2 weeks of therapy and can be minimized with use of the cream formulation, alternate day application, short contact therapy, mild cleansers, and combination therapy. PMID- 12627998 TI - Cardiac function during mental stress: cholinergic modulation with pyridostigmine in healthy subjects. AB - Mentally or emotionally stressful situations occur throughout our lives and cause physiological haemodynamic responses. In patients with coronary artery disease, such events can also induce myocardial ischaemia and ventricular arrhythmias, increasing mortality rates. The purpose of the present study was to determine the acute effects of the oral administration of pyridostigmine, a reversible cholinesterase inhibitor and thus an indirect cholinomimetic drug, on echocardiographic variables during mental stress in healthy subjects. A total of 18 healthy young volunteers were subjected to mental stress tests (mental arithmetic and the Stroop colour-word test) 2 h after the oral administration of either placebo or pyridostigmine bromide (45 mg), using a balanced-randomized, double-blind, crossover protocol. During mental stress, heart rate (pyridostigmine, 64+/-1 beats/min; placebo, 70+/-1 beats/min; P =0.0003) and diastolic blood pressure (pyridostigmine, 66+/-2 mmHg; placebo, 79+/-3 mmHg; P =0.01) were lower in the pyridostigmine group, but systolic pressure was not (pyridostigmine, 124+/-3 mmHg; placebo, 123+/-3 mmHg; P =0.40). There were no detectable abnormalities in the left ventricular wall motion score during mental stress, but left ventricular outflow tract mean velocity (pyridostigmine, 0.68+/ 0.02 m/s; placebo, 0.64+/-0.02 m/s; P <0.05) and mitral inflow velocity deceleration (placebo, 4.05+/-0.18 m/s(2); pyridostigmine, 4.41+/-0.16 m/s(2); P <0.05) were higher in the pyridostigmine group. In conclusion, cholinergic stimulation with pyridostigmine seems to increase left ventricular diastolic function during mental stress in healthy subjects. PMID- 12627996 TI - Topical 3% diclofenac in 2.5% hyaluronic acid gel: a review of its use in patients with actinic keratoses. AB - Topical 3% diclofenac in 2.5% hyaluronic acid (HA) gel (diclofenac HA gel; Solaraze) is an NSAID approved for the treatment of actinic keratoses (AK). The efficacy of diclofenac HA gel (0.5g applied twice daily to each 5cm x 5cm treatment area) in patients with AK has been evaluated in three randomized, double-blind, HA gel vehicle-controlled trials. In each trial, efficacy was assessed 30 days after the end of treatment because an earlier study revealed that resolution of lesions was greater when measured after a 4 week interval, rather than at the end of treatment. In two fully published multicenter trials, there was no difference in baseline characteristics of the study groups. In a further single center study (not yet published), patients randomized to diclofenac HA gel had a significantly higher mean number of target lesions at baseline compared with HA gel vehicle. In the two published studies, the efficacy of diclofenac HA gel increased with increased treatment duration. When compared with HA gel vehicle recipients, significant improvements in total lesion number scores (TLNS), cumulative lesion number scores (CLNS), patient global improvement indices (PGII) and investigator global improvement indices (IGII) were obtained in patients treated for 60 and 90 but not 30 days with diclofenac HA gel. Fifty percent of patients treated for 90 days with diclofenac HA gel (vs 20% in HA gel vehicle recipients) and 33% of those treated for 60 days (vs 10%) had TLNS and CLNS of zero at the end of follow-up. In the third trial, in which treatment was applied for 90 days, there was no statistically significant difference in the proportion of patients with TLNS or CLNS of zero at the end of follow-up. However, when controlling for the significant difference in mean baseline target lesion scores by calculating the mean change from baseline in lesion counts, TLNS and CLNS were significantly lower in recipients of diclofenac HA gel than HA gel vehicle at the end of follow-up. Pruritus was the most frequently reported adverse event in all trials and the incidence was generally similar or lower in patients treated with diclofenac HA gel than HA gel vehicle. In conclusion, diclofenac HA gel produces significant reductions in the number of AK lesions, and can produce complete clearance of lesions when applied twice daily for 60 or 90 days. The product is well tolerated and did not produce serious adverse cosmetic effects in clinical trials. Thus, diclofenac HA gel represents a useful addition to the array of pharmacologic treatments available for AK. PMID- 12627999 TI - Energetics of walking in patients with peripheral arterial disease: a proposed functional evaluation protocol. AB - The energy cost of walking (at 3.2 km x h(-1)) per unit distance (J x kg(-1) x m( 1)) at gradients of 0%, +7%, and +12% and during a progressive test (2% increase in gradient every 2 min), as well as the overall (aerobic plus anaerobic) net cumulative energy consumption and the corresponding maximal exercise duration were assessed in 19 patients with peripheral arterial disease (PAD) and in 13 moderately active control subjects. With a 0% gradient, the energy cost of walking was approximately 40% greater in patients with PAD than in controls (2.93+/-0.52 and 2.13+/-0.33 J x kg(-1) x m(-1) respectively; P <0.01). In contrast, at gradients of +7% and +12%, the energy cost of walking was similar in the two groups (+7%: PAD, 4.15+/-0.74 J x kg(-1) x m(-1); controls, 4.18+/-0.54 J x kg(-1) x m(-1); +12%: PAD, 5.59+/-1.03 J x kg(-1) x m(-1); controls, 5.64+/ 0.75 J x kg(-1) x m(-1)). In patients with PAD, maximal exercise duration with gradients of 0%, +7% and +12% was 449+/-254, 322+/-200 and 229+/-150 s respectively, whereas the net cumulative energy consumption at fatigue was almost constant at approximately 1100 J x kg(-1) for all gradients. The greater energy cost of walking in PAD patients compared with controls in level, but not uphill, walking is interpreted as being mainly the consequence of an altered mechanical locomotory pattern, and not of lower metabolic efficiency. For a wide range of loads, net cumulative energy consumption appears to be independent of maximal exercise duration, a finding that provides a practical criterion for assessing the degree of functional impairment of patients with PAD on metabolic grounds. PMID- 12628000 TI - Nucleotide-dependent protein folding in the type II chaperonin from the mesophilic archaeon Methanococcus maripaludis. AB - We report the characterization of the first chaperonin (Mm-cpn) from a mesophilic archaeon, Methanococcus maripaludis. The single gene was cloned from genomic DNA and expressed in Escherichia coli to produce a recombinant protein of 543 amino acids. In contrast with other known archaeal chaperonins, Mm-cpn is fully functional in all respects under physiological conditions of 37 degrees C. The complex has Mg(2+)-dependent ATPase activity and can prevent the aggregation of citrate synthase. It promotes a high-yield refolding of guanidinium-chloride denatured rhodanese in a nucleotide-dependent manner. ATP binding is sufficient to effect folding, but ATP hydrolysis is not essential. PMID- 12628002 TI - An analysis of the phosphorylation and activation of extracellular-signal regulated protein kinase 5 (ERK5) by mitogen-activated protein kinase kinase 5 (MKK5) in vitro. AB - MKK5 expressed as a glutathione S-transferase fusion protein in human embryonic kidney 293 cells activated full-length extracellular-signal-regulated protein kinase (ERK)5 (ERK5wt) as well as the isolated catalytic domain (ERK5cat) in vitro. Activation was accompanied by the phosphorylation of Thr(219) and Tyr(221), the former residue being phosphorylated preferentially. ERK5cat phosphorylated at Thr(219), but not Tyr(221), possessed 10% of the activity of the doubly phosphorylated protein towards myelin basic protein, whereas ERK5cat phosphorylated at Tyr(221) alone was much less active. Activated ERK5 phosphorylated itself at a number of residues, including Thr(28), Ser(421), Ser(433), Ser(496), Ser(731) and Thr(733). ERK5 phosphorylated at Thr(219), but not Tyr(221), phosphorylated itself at a similar rate to ERK5 phosphorylated at both Thr(219) and Tyr(221). Activated ERK5 also phosphorylated mitogen-activated protein kinase kinase 5 (MKK5) extensively at Ser(129), Ser(137), Ser(142) and Ser(149), which are located within the region in MKK5 that is thought to interact with ERK5. PMID- 12628001 TI - Antibacterial properties of the sperm-binding proteins and peptides of human epididymis 2 (HE2) family; salt sensitivity, structural dependence and their interaction with outer and cytoplasmic membranes of Escherichia coli. AB - During passage through the epididymis, sperm interact with secreted epididymal proteins that promote maturation, including the acquisition of motility and fertilization competence. Viewed previously as distinct from sperm maturation, host defence capabilities are now recognized functions of the human epididymis 2 (HE2) family of sperm-binding proteins. We analysed the potent dose and time dependent bactericidal activity of recombinant HE2alpha, HE2beta1 and HE2beta2 and found that the full-length proteins (10 microg/ml or approximately 1 microM) caused more than a 50% decrease in Escherichia coli colony forming units within 15 min. By contrast, human beta-defensin-1, at a similar concentration, required more than 90 min to exhibit similar antibacterial activity. The epididymis specific lipocalin, LCN6, failed to kill bacteria. Higher concentrations (25-100 microg/ml) of HE2 proteins and a longer duration of treatment resulted in near total inhibition of bacterial growth. The C-terminal peptides of HE2alpha, HEbeta1 and HEbeta2 proteins exhibited antibacterial activity similar to their full-length counterparts, indicating that the antibacterial activity of HE2 proteins resides in these C-terminal regions. Antibacterial activities of HE2 proteins and peptides were slightly inhibited by NaCl concentrations of up to 150 mM, while human beta-defensin-1 activity was nearly eliminated. Reduction and alkylation of disulphide bonds in HE2 proteins and their C-terminal peptides abolished their antibacterial activity. Consistent with the ability to kill bacteria, full-length HE2 proteins and C-terminal peptides caused rapid dose dependent permeabilization of outer and cytoplasmic E. coli membranes. A much longer exposure time was required for human beta-defensin-1-mediated permeabilization of membranes, suggesting a possible difference in mode of action compared with the HE2 antibacterial peptides. PMID- 12628003 TI - Characterization of the nutrient-sensing response unit in the human asparagine synthetase promoter. AB - Transcription from the human asparagine synthetase (A.S.) gene is increased in response to either amino acid (amino acid response) or glucose (endoplasmic reticulum stress response) deprivation. These two independent nutrient-sensing pathways converge on the same set of genomic cis -elements, referred to as nutrient sensing-response elements (NSREs) 1 and 2, within the A.S. promoter. The present report uses single-nucleotide mutagenesis to confirm that both NSRE-1 and NSRE-2 are absolutely required for gene activation and to identify the boundaries of each binding site. The core sequence of the NSRE-1 site is contained within nucleotides -68 to -60 and the NSRE-2 core sequence is within nucleotides -48 to 43. Through insertion or deletion of 5-10 nucleotides in the intervening sequence between NSRE-1 and NSRE-2, transient transfection studies with an A.S. promoter/reporter gene construct showed that the 11 bp distance between these two elements is critical. These results document that the optimal configuration is with both binding sites on the same side of the DNA helix, only one helical turn away from each other and the data provide support for the hypothesis that a larger multi-protein complex exists between the binding proteins for NSRE-1 and NSRE-2. The data also illustrate that the combination of NSRE-1 and NSRE-2, referred to as the nutrient-sensing response unit (NSRU), has enhancer activity in that it functions in an orientation- and position-independent manner, and conveys nutrient-dependent transcriptional control to a heterologous promoter. PMID- 12628004 TI - The human neuroendocrine thyrotropin-releasing hormone receptor promoter is activated by the haematopoietic transcription factor c-Myb. AB - Thyrotropin-releasing hormone (TRH) receptor (TRHR) is a G-protein-coupled receptor playing a crucial role in the anterior pituitary where it controls the synthesis and secretion of thyroid-stimulating hormone and prolactin. Its widespread presence not only in the central nervous system, but also in peripheral tissues, including thymus, indicates other important, but unknown, functions. One hypothesis is that the neuropeptide TRH could play a role in the immune system. We report here that the human TRHR promoter contains 11 putative response elements for the haematopoietic transcription factor c-Myb and is highly Myb-responsive in transfection assays. Analysis of Myb binding to putative response elements revealed one preferred binding site in intron 1 of the receptor gene. Transfection studies of promoter deletions confirmed that this high affinity element is necessary for efficient Myb-dependent transactivation of reporter plasmids in CV-1 cells. The Myb-dependent activation of the TRHR promoter was strongly suppressed by expression of a dominant negative Myb Engrailed fusion. In line with these observations, reverse transcriptase PCR analysis of rat tissues showed that the TRHR gene is expressed both in thymocytes and bone marrow. Furthermore, specific, high-affinity TRH agonist binding to cell surface receptors was demonstrated in thymocytes and a haematopoietic cell line. Our findings imply a novel functional link between the neuroendocrine and the immune systems at the level of promoter regulation. PMID- 12628007 TI - Expression of the Bacillus thuringiensis Cyt2Aa1 toxin in Pichia pastoris using a synthetic gene construct. AB - The nucleotide sequence data corresponding to the syncyt2Aa1 open reading frame was deposited with the EMBL/GenBank Nucleotide Sequence Databases under the accession number AF398463. Bacillus thuringiensis delta-endotoxins are membrane active, pore-forming proteins with highly specific insecticidal activities. In addition to a well-established role in the biological control of a wide variety of crop pests and disease vectors, these toxins also have great potential for the development of anti-tumour agents called immunotoxins (ITs), chimaeric molecules consisting of a cell-binding ligand coupled to a toxin or its subunits. The ultimate goal of our study was the recombinant production of such ITs based on the Cyt2Aa1 toxin from B. thuringiensis subspecies kyushuensis. We explored the use of Pichia pastoris for recombinant IT production because earlier attempts in our laboratory using the Escherichia coli expression system or various chemical conjugation strategies yielded only low levels of functional product. However, our initial attempts were not successful because the A+T-rich bacterial cyt2Aa1 gene contained fortuitous polyadenylation sites, causing premature transcription termination in this yeast. Accordingly, we designed and constructed a synthetic cyt2Aa1 gene (syncyt2Aa1) optimized for heterologous expression in P. pastoris. This was achieved by increasing the overall G+C content of the bacterial cyt2Aa1 while changing its codon usage to that preferred by the methylotrophic yeast. Here we describe in detail the design, synthesis and requisite PCR repair of syncyt2Aa1, then present analyses of recombinant Cyt2Aa1 expression in P. pastoris using this synthetic gene. Following the results presented in this paper, the syncyt2Aa1 gene was also successfully used for the recombinant production of a Cyt2Aa1-based IT in the same expression host. PMID- 12628019 TI - Glucocorticoid-like effects of antihepatocarcinogen Rotenone are mediated via enhanced serum corticosterone levels: Molecular Fitting and Receptor Activation Studies. AB - BACKGROUND: Recent studies suggest that rotenone alters cell signal transduction pathways in a manner similar to glucocorticoids. Histological and biochemical markers of glucocorticoid effects in vivo, evaluated in our laboratories, provide further evidence for similarities in the activity of glucocorticoids and rotenone. The purpose of this study was to investigate the mechanism by which rotenone produces glucocorticoid-like effects. METHODS: Male B6C3F1 mice were treated for 7 days with rotenone (600 ppm in diet), the glucocorticoid antagonist RU486 (2 mg/kg/day, ip), corticosterone (2 mg/kg/day, ip), or both rotenone and RU 486. Control mice received drug-free diet and the vehicle (corn oil, ip). Following preservation in 10% neutral buffered formalin, tissues were embedded in paraffin. Sections were stained with hematoxylin, eosin, and were examined by light microscopy. Tissue sections were processed for in situ enzymatic end labeling of 3'-hydroxy-DNA strand breaks, a measure of apoptosis. Corticosterone was quantified in sera, using a solid phase radioimmunoassay kit. Cells (cell line 1470.2 derived from C127 mouse mammary adenocarcinoma cells) were transiently transfected with 5 &mgr;g of pLTRLuc and 1 &mgr;g of beta Galactosidase expression vectors using a BTX square-wave pulser at 155 V, 4 pulses (40 ms each). Cells were then treated with dexamethasone, rotenone, or a mixture of both for 6 hr, harvested and assayed for luciferase and beta Galactosidase activity. Using Root Mean Square (RMS) fit analysis (Alchemy trade mark, Tripose, Inc., St Louis, MO), we assessed possible structural similarities between rotenone and corticosterone, dehydrocorticosterone, glucocorticoid antagonists ZK 98.299, and RU 486. RMS fit was calculated by selecting three atoms in each of the molecules, followed by calculating the distance between these atoms. An RMS value of zero between two molecules indicates identical molecular characteristics. A positive value suggests diminished similarity with a value of 1 or higher excluding any such similarities. RESULTS: Although the stimulatory effect exerted by rotenone on hepatocellular apoptosis was in the opposite direction of that produced by the glucocorticoid antagonist RU 486, data suggested that rotenone does not directly activate the glucocorticoid receptor. Molecular fitting of rotenone to glucocorticoid receptor agonists and antagonists as well as examination of the transcriptional activation of a glucocorticoid responsive reporter gene (Mouse MammaryTumorVirus) in response to rotenone indicated that it is highly unlikely that rotenone interacts directly with the glucocorticoid receptor. However, feeding male B6C3F1 mice a diet containing rotenone (600 ppm for 7 days) resulted in a 3-fold increase in serum levels of corticosterone relative to control animals. Corticosterone is the major glucocorticoid in rodents. CONCLUSION: Rotenone does not interact directly with the glucocorticoid receptor. Elevation of serum corticosterone levels in response to rotenone may explain the glucocorticoid-like effects of this compound, and may play a role in its anti-hepatocarcinogenic effect. PMID- 12628005 TI - Ets2 and protein kinase C epsilon are important regulators of parathyroid hormone related protein expression in MCF-7 breast cancer cells. AB - Parathyroid hormone-related protein (PTHrP) promotes the metastatic potential and proliferation of breast cancer cells, and acts anti-apoptotically. In invasive MDA-MB-231 breast cancer cells, transforming growth factor beta-regulated PTHrP synthesis is mediated by an Ets1/Smad3-dependent activation of the PTHrP P3 promoter. In the present study, we studied the regulation of PTHrP expression in non-invasive, Ets1-deficient and transforming growth factor beta-resistant MCF-7 cells. We found PMA to be a strong stimulator of P3-dependent PTHrP expression in MCF-7 cells. Mitogen-activated protein kinase (MAPK)/extracellular-signal regulated kinase (ERK) kinase 1 (MEK-1)/ERK1/2 inhibitor PD98059 interfered with this activity. Promoter studies revealed that the PMA effect depended on the Ets and stimulating protein-1 (Sp1)-binding sites. Of several Ets factors tested, Ets2, but not Ese-1, Elf-1 or Ets1, supported the PMA-dependent increase in promoter activity. PD98059 and a threonine to alanine mutation of the ERK1/2 responsive Ets2 phosphorylation site at position 72 inhibited the Ets2/PMA effect. Activated protein kinase C (PKC) epsilon could mimic PMA by stimulating the P3 promoter alone or in co-operation with Ets2 in an MEK-1/ERK1/2-dependent manner. Activated PKC alpha, although capable of co-operating with Ets2, failed to induce transcription from the P3 promoter on its own. The Ets2/PKalpha synergistic effect was neither sensitive to PD98059 nor to Thr(72)/Ala(72) mutation. PMA neither increased the expression of Sp1 nor modulated the transcriptional activity of Sp1. However, it induced the displacement of a yet unknown factor from the Sp1-binding site, which may result in Sp1 recruitment to the promoter. Our results suggest an ERK1/2-dependent Ets2/PKC epsilon synergism to be involved in PTHrP expression in MCF-7 breast cancer cells. PMID- 12628006 TI - Tyrosine phosphorylation of protein kinase CK2 by Src-related tyrosine kinases correlates with increased catalytic activity. AB - Casein kinase-2 (CK2) is a pleiotropic and constitutively active serine/threonine protein kinase composed of two catalytic (alpha and/or alpha') and two regulatory beta-subunits, whose regulation is still not well understood. In the present study, we show that the catalytic subunits of human CK2, but not the regulatory beta-subunits, are readily phosphorylated by the Src family protein tyrosine kinases Lyn and c-Fgr to a stoichiometry approaching 2 mol phosphotyrosine/mol CK2alpha with a concomitant 3-fold increase in catalytic activity. We also show that endogenous CK2alpha becomes tyrosine-phosphorylated in pervanadate-treated Jurkat cells. Both tyrosine phosphorylation and stimulation of activity are suppressed by the specific Src inhibitor 4-amino-5-(4-chlorophenyl)-7-(t butyl)pyrazolo[3,4- d ]pyrimidine. By comparison, mutations giving rise to inactive forms of CK2alpha do not abrogate and, in some cases, stimulate Lyn and c-Fgr-dependent tyrosine phosphorylation of CK2. Several radiolabelled phosphopeptides could be resolved by HPLC, following tryptic digestion of CK2alpha that had been phosphoradiolabelled by incubation with [(32)P]ATP and c Fgr. The most prominent phosphopeptide co-migrates with a synthetic peptide encompassing the 248-268 sequence, phosphorylated previously by c-Fgr at Tyr(255) in vitro. The identification of Tyr(255) as a phosphorylated residue was also supported by MS sequencing of both the phosphorylated and non-phosphorylated 248 268 tryptic fragments from CK2alpha and by on-target phosphatase treatment. A CK2alpha mutant in which Tyr(255) was replaced by phenylalanine proved less susceptible to phosphorylation and refractory to stimulation by c-Fgr. PMID- 12628021 TI - Noninvasive assessment of vascular function in the posterior tibial artery of healthy humans. AB - Flow-mediated dilation (FMD) measures the ability of an artery to relax in response to increases in blood velocity. FMD, primarily of the brachial artery, has been used as a noninvasive method of assessing vascular health. The purpose of this study was to assess FMD in the lower legs of humans. Six healthy subjects (27 PlusMinus; 6 yrs) were tested. Doppler ultrasound images of the posterior tibial artery were taken before, during, and after 5 minutes of proximal cuff occlusion. FMD was measured as the percent increase in diameter after cuff release. Vascular tone was calculated using the resting diameter as a percentage of the vessel's vasoactive range. Minimum diameter occurred during ischemia and maximal diameter occurred following reactive hyperemia with local heating. The lower leg was heated with 10 minutes of immersion in 44 degrees C water. Mean diameters at rest, cuff, and during release were 0.267 PlusMinus; 0.062, 0.162 PlusMinus; 0.036, 0.302 PlusMinus; 0.058 cm, respectively. FMD was 13.5 PlusMinus; 6.6 % and vascular tone was 29 PlusMinus; 16.3%. We also found that retesting on a second day produced mean diameter values within 8% of the first day. Larger resting diameter (decreased tone) correlated with decreased FMD (r2 = 0.73). These results suggest that FMD and vascular tone can be measured in the posterior tibial artery. This is a potentially powerful tool to non-invasively measure vascular health in the lower legs of people at risk for vascular disease. PMID- 12628020 TI - The HIV vaccine saga. AB - The development of a vaccine that can prevent infection by the Human immunodeficiency virus or prevent the Acquired Immunodeficiency Syndrome has remained elusive despite 20 years of scientific effort. This "Commentary" analyzes the reasons that the development of a vaccine has been so difficult, and proposes a plan to work towards an immunological approach to investigate the best vaccine candidates in the first world in individuals who are already infected, before taking the most promising vaccines to the developing world to attempt to prevent infection and disease. SAGA: (Old Norse) "a long, continued heroic story that is action-packed, but not especially romantic, and that is historical or legendary or both". PMID- 12628022 TI - Is type 2 diabetes mellitus a vascular disease (atheroscleropathy) with hyperglycemia a late manifestation? The role of NOS, NO, and redox stress. AB - BACKGROUND: Cardiovascular disease accounts for at least 85 percent of deaths for those patients with type 2 diabetes mellitus (T2DM). Additionally, 75 percent of these deaths are due to ischemic heart disease. HYPOTHESIS: Is type 2 diabetes mellitus a vascular disease (atheroscleropathy) with hyperglycemia a late manifestation? The role of NOS, NO, and redox stress. TESTING OF THE HYPOTHESIS: The vulnerable three arms of the eNOS reaction responsible for the generation of eNO is discussed in relation to the hypothesis: (1) The L-arginine substrate. (2) The eNOS enzyme. (3) The BH4 cofactor. IMPLICATIONS OF THE HYPOTHESIS: If we view T2DM as a vascular disease initially with a later manifestation of hyperglycemia, we may be able to better understand and modify the multiple toxicities associated with insulin resistance, metabolic syndrome, prediabetes, overt T2DM, and accelerated atherosclerosis (atheroscleropathy). The importance of endothelial nitric oxide synthase, endothelial nitric oxide, tetrahydrobiopterin (BH4), L arginine, and redox stress are discussed in relation to endothelial cell dysfunction and the development and progression of atheroscleropathy and T2DM. In addition to the standard therapies to restore endothelial cell dysfunction and stabilization of vulnerable atherosclerotic plaques, this article will discuss the importance of folic acid (5MTHF) supplementation in this complex devastating disease process. Atheroscleropathy and hyperglycemia could be early and late manifestations, respectively, in the natural progressive history of T2DM. PMID- 12628023 TI - The problem of predicting food intake during the period of adaptation to a new food: a model. AB - A model is described which aims to predict intake immediately following a change from one food to another that is higher in bulk content; it deals with the transition from one 'equilibrium' intake to another. The system considered is an immature pig fed ad libitum on a single homogeneous food, which is balanced for nutrients and contains no toxins so that the first limiting resource is always energy. It is assumed that an animal has a desired rate of food intake (DFI) which is that needed to meet the energy requirements for protein and lipid deposition and for maintenance. DFI may not be achieved if a bulk constraint to intake exists. Where a bulk constraint operates intake is calculated as constrained food intake (CFI) where (where WHC is the water-holding capacity of the food (kg water/kg dry food) and Cwhc is the animal's capacity for WHC (units/kg live weight per d)). Where intake is not constrained it is assumed that genetic potential will be achieved. Potential growth rate is described by the Gompertz growth function. Where intake is constrained, growth will be less than the potential. Constrained growth rate is predicted as where W is pig weight (kg), EI is energy intake (MJ/d), Em is the energy required for maintenance (MJ/d) and eg is the energy required for unit gain (MJ/kg). The value of eg depends on weight and the fattening characteristics of the pig. Actual growth is predicted to be the lesser of potential and constrained growth. To deal with adaptation it is assumed that the time taken to reach equilibrium depends on the difference in WHC values between the previous and current food and that the capacity to consume food bulk is related to the WHC of the current food. It is proposed that the capacity for WHC on the first day on a new food will be equal to the current capacity for WHC on the last day of the previous food. Thus where FI is food intake (kg/d). Thereafter Cwhc will gradually increase over time to a maximum of 0.27 g/kg. The rate of change in Cwhc is made to be the same for all pigs and all foods. The increase in capacity over time is assumed to be linear at the rate of 0.01 units/d. The model was tested using published data. Qualitatively the predictions of the model were in close agreement with the relevant observed data in at least some cases. It is concluded that the underlying theoretical assumptions of the model are reasonable. However, the model fails to predict initial intake when changed to foods high in wheat-bran content and fails to predict the intake of a non-limiting food where compensatory increases in intake and gain occur. The model could be adapted to overcome the first failure by taking into account the time course of digestive efficiency following a change in food. To deal with the second would require a sufficient understanding of the time course of compensatory growth. PMID- 12628024 TI - Hydrolysis of phytic acid and its availability in rabbits. AB - Twenty weaned rabbits were fed ad libitum two granulated feeds containing lucerne meal, barley, oats, wheat bran, oilseed meals and sugarbeet pulp in different proportions. Phytate P in these feeds represented 28.6 and 29.3 % of the total P. Digestibility trials were carried out in rabbits 7 and 10 weeks old. Digestibility of phytate P was 82.1 %, on average. Apparent digestibility of total P was 48.1 and 35.5 % in rabbits aged 7 and 10 weeks, respectively. Concentration of P in the faecal DM of these rabbits averaged 11.9 and 14.7 mg/g. Most of the faecal P was phosphates P (68.1 %). Proportion of phytate P in total faecal P was 9.0 %. Age effect on total P digestibility and faecal P concentration was significant (P<0.05). In five in vitro experiments twenty-four rabbits were killed at the age of 11 weeks, digesta samples diluted with physiological saline containing phytic acid and incubated anaerobically. Calculations of phytase activity in segments of the digestive tract were based on the estimation of phytic acid hydrolysed during the first 2 h of incubation. The caecum contained 58.6 % of the phytase activity of the digestive tract. Corresponding relative values for the phytase activity in the stomach, small intestine and colon were 22.3, 7.7 and 11.4 %, respectively. In incubations of the caecal contents, phytic acid was hydrolysed more rapidly at pH 5-6 than in the neutral pH region. The hydrolysis was inhibited by Ca cations, and to a small extent also by phosphate anions. Commercial fungal phytase (Natupho) was highly active in incubations of the contents of the stomach at pH 1.9. It can be concluded that phytic acid is hydrolysed quite efficiently in the digestive tract of rabbits. This hydrolysis occurred mainly in the caecum. Absorption of soluble inorganic phosphates in the gut is incomplete. PMID- 12628025 TI - Comparison of vitamin C deficiency with food restriction on collagen cross-link ratios in bone, urine and skin of weanling guinea-pigs. AB - Mild-to-moderate vitamin C depletion in weanling guinea-pigs affects pyridinoline:deoxypyridinoline (collagen cross-link) ratios in femur shaft and urine, attributed to impairment of hydroxylation of collagen lysine. We investigated: (1). whether the picture at two time points is compatible with progressive accumulation of abnormal collagen; (2). whether any changes are seen in skin, where little deoxypyridinoline occurs; (3). whether total food restriction has similar effects. Male weanling Dunkin-Hartley guinea-pigs were fed diets containing either 0.5 (vitamin C-restricted) or 160.0-320.0 (vitamin C adequate) mg vitamin C/d. Two groups receiving the vitamin C-adequate diet received it ad libitum. Two other groups received the vitamin C-adequate diet in a restricted amount, limited to that which permitted nearly the same growth rate as in the vitamin C-restricted groups. Animals were fed for 4 or 8 weeks; urine was collected, and vitamin C and collagen indices were measured. In the femur shaft, the hydroxyproline content per unit weight was unaffected by vitamin C restriction or by total food restriction. Deoxypyridinoline was increased and the pyridinoline:deoxypyridinoline ratio was decreased in vitamin C-restricted groups, but not in food-restricted groups. Changes in the value of the ratio were greater after 8 than after 4 weeks. Urine indices mirrored bone indices. In skin, the main effect of vitamin C restriction was to reduce hydroxyproline. Here, the cross-link ratios changed less markedly than in bone, and there was less deoxypyridinoline. We conclude that the picture at two time points is compatible with a progressive accumulation of pyridinoline-enriched collagen in vitamin C deprived animals, that the picture in skin differs from that of bone and urine, and that cross-link changes are not produced by total food restriction. PMID- 12628026 TI - Local (gut) and systemic metabolism of rats is altered by consumption of raw bean (Phaseolus vulgaris L var athropurpurea). AB - The composition of the raw legume Phaseolus vulgaris L. var. athropurpurea (PhVa) and its effects on the metabolism of young growing rats have been evaluated. The levels of protein, unsaturated fatty acids, carbohydrate, fibre and bioactive factors present in PhVa were comparable with those in other Phaseolus vulgaris varieties. However, the lectins of PhVa were predominantly of the leucoagglutinating type, and concentrated in the albumin protein fraction. Rats fed a diet (110 g total protein, 16.0 MJ/kg) in which PhVa meal provided about half of the protein excreted high levels of N in faeces and urine, and grew more slowly, than rats fed a high-quality control diet (ad libitum or pair-fed). Small intestine, large intestine and pancreas weights were increased (by almost 100 %, P<0.05), whilst skeletal muscle, thymus and spleen weights were reduced. Blood insulin (16.20 v. 0.50 mU/l, P<0.05, thyroxine, glucose, protein (60.5 v. 48.3 g/l, P<0.05) and LDL-cholesterol were lowered, whilst glucagon (155.3 v. 185.4 ng/l, P<0.05), triiodothyronine and urea were elevated, as were urinary urea, creatinine and glucose. These changes in the local (gut) and systemic metabolism of rats were probably mediated primarily by lectins in PhVa, which were concentrated in the albumin protein fraction, whereas in many other Phaseolus vulgaris lines they are distributed across the globulin and albumin fractions. PMID- 12628027 TI - Influences of dietary adaptation and source of resistant starch on short-chain fatty acids in the hindgut of rats. AB - The effect of adaptation time on the concentration and pattern of short-chain fatty acids (SCFA) formed in the hindgut of rats given resistant starch (RS) in the form of raw potato starch (RPS) or high-amylose maize starch (HAS) was evaluated. Each starchy material was tested in diets containing 100 g indigestible carbohydrates/kg DM, and fed for 13, 28 and 42 d. At the end of each period, the content of SCFA was determined in caecum, distal colon and faeces. The caecal concentration of total and individual SCFA increased for both diets with increasing adaptation time. The concentration of butyric acid was higher in the group fed RPS than in that fed HAS at all adaptation times. The caecal proportion of butyric acid was low both in rats fed RPS and HAS (6 and 4 %, respectively) following 13 d of adaptation. However, after 28 d of adaptation, the proportion of butyric acid had increased to 19 % in rats given RPS. A longer adaptation period (42 d) did not increase the proportion of butyric acid further. With HAS, there was also a significant (P<0.01) increase in the proportion of butyric acid with longer adaptation time. However, the increase was much slower and the proportion of butyric acid reached 6 and 8 % after 28 and 42 d respectively. It is concluded that the pattern of SCFA formed from RS in rats is dependent on adaptation time. It cannot be excluded that the different patterns of SCFA reported in the literature for RS may be due to the time of adaptation. PMID- 12628028 TI - Glycaemic index of cereal products explained by their content of rapidly and slowly available glucose. AB - Elucidating the role of carbohydrate quality in human nutrition requires a greater understanding of how the physico-chemical characteristics of foods relate to their physiological properties. It was hypothesised that rapidly available glucose (RAG) and slowly available glucose (SAG), in vitro measures describing the rate of glucose release from foods, are the main determinants of glycaemic index (GI) and insulinaemic index (II) for cereal products. Twenty-three products (five breakfast cereals, six bakery products and crackers, and twelve biscuits) had their GI and II values determined, and were characterised by their fat, protein, starch and sugar contents, with the carbohydrate fraction further divided into total fructose, RAG, SAG and resistant starch. Relationships between these characteristics and GI and II values were investigated by regression analysis. The cereal products had a range of GI (28-93) and II (61-115) values, which were positively correlated (r(2)) 0.22, P<0.001). The biscuit group, which had the highest SAG content (8.6 (SD 3.7) g per portion) due to the presence of ungelatinised starch, was found to have the lowest GI value (51 (SD 14)). There was no significant association between GI and either starch or sugar, while RAG was positively (r(2)) 0.54, P<0.001) and SAG was negatively (r(2)) 0.63, P<0.001) correlated with GI. Fat was correlated with GI (r(2)) 0.52, P<0.001), and combined SAG and fat accounted for 73.1% of the variance in GI, with SAG as the dominant variable. RAG and protein together contributed equally in accounting for 45.0 % of the variance in II. In conclusion, the GI and II values of the cereal products investigated can be explained by the RAG and SAG contents. A high SAG content identifies low-GI foods that are rich in slowly released carbohydrates for which health benefits have been proposed. PMID- 12628030 TI - Detection of caseinophosphopeptides in the distal ileostomy fluid of human subjects. AB - Caseinophosphopeptides (CPP) were detected for the first time in ileostomy fluid, collected at 2 h intervals for 10 h post milk and CPP ingestion, from human volunteers with an ileostomy. The level of CPP present in ileostomy fluid obtained from milk-fed volunteers was markedly higher than that from volunteers fed with selected CPP preparations. The findings are based on HPLC analysis in combination with peptide-bound P determination, thin-layer electrophoresis and amino acid analysis, together with ELISA studies using polyclonal antibodies raised against a set of CPP to detect immunoreactive CPP in ileostomy fluid. These procedures allowed the detection of nm concentrations of CPP. CPP, which can be released during intestinal digestion, may function as bioactive constituents and carriers for different minerals, especially Ca, and may be used as ingredients in functional foods or pharmaceutical preparations. PMID- 12628029 TI - Dietary cholesterol reduces lipoprotein lipase activity in the atherosclerosis susceptible Bio F(1)B hamster. AB - We have compared lipoprotein metabolism in, and susceptibility to atherosclerosis of, two strains of male Golden Syrian hamster, the Bio F(1)B hybrid and the dominant spot normal inbred (DSNI) strain. When fed a normal low-fat diet containing approximately 40 g fat and 0.3 g cholesterol/kg, triacylglycerol-rich lipoprotein (chylomicron+VLDL) and HDL-cholesterol were significantly higher (P<0.001) in Bio F(1)B hamsters than DSNI hamsters. When this diet was supplemented with 150 g coconut oil and either 0.5 or 5.0 g cholesterol/kg, significant differences were seen in response. In particular, the high cholesterol diet produced significantly greater increases in plasma cholesterol and triacylglycerol in the Bio F(1)B compared with the DSNI animals (P=0.002 and P<0.001 for cholesterol and triacylglycerol, respectively). This was particularly dramatic in non-fasting animals, suggesting an accumulation of chylomicrons. In a second experiment, animals were fed 150 g coconut oil/kg and 5.0 g cholesterol/kg for 6 and 12 months. Again, the Bio F(1)B animals showed dramatic increases in plasma cholesterol and triacylglycerol, and this was confirmed as primarily due to a rise in chylomicron concentration. Post-heparin lipoprotein lipase activity was significantly reduced (P<0.001) in the Bio F(1)B compared with the DSNI animals at 6 months, and virtually absent at 12 months. Bio F(1)B animals were also shown to develop significantly more (P<0.001) atherosclerosis. These results indicate that, in the Bio F(1)B hybrid hamster, cholesterol feeding reduces lipoprotein lipase activity, thereby causing the accumulation of chylomicrons that may be associated with their increased susceptibility to atherosclerosis. PMID- 12628031 TI - Folic acid treatment reduces elevated plasma levels of asymmetric dimethylarginine in hyperhomocysteinaemic subjects. AB - Asymmetric dimethylarginine (ADMA), an endogenous inhibitor of NO synthase, has been suggested to be a novel risk factor for endothelial dysfunction. It has previously been reported that hyperhomocysteinaemia may be associated with impaired endothelium-dependent vasodilation and reduced plasma level of NO derived endproducts (NOx). In the present study, plasma levels of arginine and ADMA were measured in twenty-one healthy control subjects, and in twenty-one hyperhomocysteinaemic subjects before and after 6 weeks and 12 months of folic acid supplementation, and compared with previously measured plasma NOx values in the hyperhomocysteinaemic subjects. Compared with control subjects, hyperhomocysteinaemic subjects had higher plasma levels of arginine and ADMA. More importantly, folic acid therapy significantly reduced plasma levels of arginine and ADMA. Furthermore, plasma levels of arginine and ADMA were positively correlated with plasma homocysteine levels and negatively correlated with plasma folate, as well as negatively correlated with plasma NOx. Our results suggest that ADMA may be a mediator of the atherogenic effects of homocysteine. PMID- 12628032 TI - Carbohydrate-induced manipulation of insulin sensitivity independently of intramyocellular lipids. AB - Subjects with insulin resistance have been shown to have higher storage levels of intramyocellular lipid (IMCL) than their insulin-sensitive counterparts. It has been proposed that elevated IMCL stores may be the main cause of insulin resistance. The aim of the present study was to ascertain whether there is a causal relationship between IMCL storage and insulin resistance. IMCL storage was assessed using magnetic resonance spectroscopy and insulin sensitivity was assessed by performing an oral glucose tolerance test. A 4-week intervention of reduction of dietary glycaemic index was used to manipulate insulin sensitivity in a cohort of healthy volunteers; the effects of this intervention on IMCL were measured after 4 weeks of intervention. Significant improvements in the insulin sensitivity index occurred following the dietary intervention (baseline 7.8 (sem 1.11) v. post-intervention 9.7 (sem 1.11), P=0.02). However, there were no changes in IMCL storage levels, suggesting that insulin sensitivity can be manipulated independently of IMCL. This suggests that in healthy volunteers, insulin sensitivity is independent of IMCL storage and the high storage levels that have been found in insulin-resistant subjects may occur as a consequence rather than a cause of insulin resistance. PMID- 12628033 TI - The source of fatty acids incorporated into proliferating lymphoid cells in immune-stimulated lymph nodes. AB - To explore the hypothesis that proliferating lymphoid cells in immune-stimulated lymph nodes obtain nutrients locally from adjacent adipose tissue, adult guinea pigs were fed for 6 weeks on standard chow or on chow supplemented with 100 g suet, sunflower oil or fish oil/kg. All the guinea pigs ate standard chow for the last 5 d, during which swelling of one popliteal lymph node was stimulated by repeated local injection of lipopolysaccharide. The fatty acid compositions of phospholipids in both popliteal and in several mesenteric lymph nodes, and of triacylglycerols in eleven samples of adipose tissue defined by their anatomical relations to lymph nodes, were determined by GC. The proportions of fatty acids in the phospholipids extracted from the stimulated popliteal node correlated best with those of triacylglycerols in the surrounding adipocytes, less strongly with those of adipocytes elsewhere in depots associated with lymphoid tissue, but not with those of nodeless depots. The composition of triacylglycerols in the perinodal adipose tissue changed under local immune stimulation. We conclude that proliferating lymphoid cells in activated lymph nodes obtain fatty acids mainly from the triacylglycerols in adjacent perinodal adipose tissue. Immune stimulation prompts changes in the fatty acid composition of the triacylglycerols of adipocytes in node-containing depots that equip the adipose tissue for provisioning immune responses. Such local interactions show that specialised adipocytes can act as an interface between whole-body and cellular nutrition, and may explain why mammalian adipose tissue is partitioned into a few large and many small depots. PMID- 12628034 TI - Effects of altitude, ambient temperature and solar radiation on fasting heat production in yellow cattle (Bos taurus). AB - Growing yellow cattle (Bos taurus, n 30, 1.0-3.5 years old and 75-240 kg) from their native altitude (2000-2800 m) were used to evaluate the effects of altitude, ambient temperature (Ta) and solar radiation on the basal energy metabolism in this large mammal. Fasting heat production (FHP) was measured at altitudes of 2260, 3250 and 4270 m on the Tibetan plateau both in the summer and winter respectively, after a 90 d adaptation period at each experimental site. The gas exchanges of the whole animal were determined continuously for 3 (2260 and 3250 m) or 2 (4270 m) d after a 96 (2260 and 3250 m) or 48 (4270 m) h starvation period, using closed-circuit respiratory masks. Increasing altitude from 2260 to 3250 m at similar Ta in the summer significantly elevated FHP for all animals (P<0.01), and from 3250 to 4270 m for young cattle (P<0.05); increasing altitude from 2260 to 3250 m in the winter also significantly elevated FHP (P<0.05), but the increase was mainly due to the decrease of Ta and the increase in wind speed. No results were obtained at 4270 m in the winter, due to the problems of the animals, adapting to the altitude. The magnitude of FHP elevation caused by increasing altitude was greater with summer sunshine or winter wind than without them. Increase of Ta from 10.0 to 22.0 degrees C, in the presence of solar radiation, slightly (2260 m) or significantly (3250 and 4270 m, P<0.01) elevated FHP, but slightly reduced it in the absence of solar radiation; decrease of Ta from 0.0 to -30.0 degrees C linearly increased FHP. At 3250 and 4270 m, FHP at the same Ta was higher with summer sunshine or winter wind (3250 m) than without them, but this did not occur at 2260 m. In conclusion, high altitude elevates FHP in yellow cattle in the warm season, and the summer solar radiation and winter wind at high altitude significantly increase metabolic rate. It may be also concluded that the effects of solar radiation on metabolic rate depend on the altitude and the environmental temperature. PMID- 12628035 TI - An approach to construct simplified measures of dietary patterns from exploratory factor analysis. AB - Exploratory factor analysis might work well in elucidating the major dietary patterns prevailing in specific study populations. However, patterns extracted in one study population and their associations with disease risk cannot be reproduced with this data-specific method in other study populations. To construct less population-dependent pattern variables of similar content as original exploratory patterns, we proposed to derive so-called simplified pattern variables. They represent the sum of the unweighted standardised food variables which loaded high at the pattern of interest. Data from the European Prospective Investigation into Cancer and Nutrition (EPIC)-Potsdam study suggest that these simplified pattern variables might adequately approximate factor analysis-based dietary patterns. A simplified pattern variable based on the six highest loading food variables showed a correlation >0.95 with the originally derived factor score, which consisted of forty-seven food variables. Moreover, simplified pattern variables might adequately approximate patterns across different study populations. A simplified pattern variable showed similar factor loadings, ranging from 0.34 to 0.52, as well as similar associations with nutrient intake as a 'western' pattern originally reported from an US study population. These simplified pattern variables can subsequently be used to study pattern associations with disease risk, especially in multi-centre studies. It is therefore an approach that might overcome one of the most frequently claimed limitations of factor analyses applied in epidemiology: their non-comparable risk estimates. PMID- 12628036 TI - Consumption of bakery products, sweetened soft drinks and yogurt among children aged 6-7 years: association with nutrient intake and overall diet quality. AB - The present study tests the hypothesis that higher consumption of bakery products, sweetened soft drinks and yogurt is associated with higher intake of energy, saturated fats, sugars and worse overall diet quality among Spanish children. This is a cross-sectional study covering 1112 children aged 6.0-7.0 years in four Spanish cities. Nutrient and food intake were obtained through a food-frequency questionnaire, and overall diet quality calculated using the healthy-eating index (HEI) developed by Kennedy et al. (1995). Standardized methods were used to measure anthropometric variables. Associations of interest were summarized as the difference in nutrient and food consumption between the value of the fifth and the first quintile of consumption (dq) of bakery products, sweetened soft drinks or yogurt, adjusted for energy intake and BMI. Bakery products, sweetened soft drinks and yogurt supplied 15.5, 1.0 and 5.6 % energy intake respectively. Higher consumption of these three foods was associated with greater energy intake (P<0.001), but not with higher BMI. Consumption of bakery products was associated with the proportion of energy derived from intake of total carbohydrates (dq 4.5 %, P<0.001) and sugars (dq 2 %, P<0.001), but did not show association with the HEI. Consumption of sweetened soft drinks was associated with a lower consumption of milk (dq -88 ml, P<0.001) and Ca (dq -175 mg/d, P<0.001), and worse HEI (dq -2, P<0.01). Consumption of yogurt, while associated with higher energy intake from saturated fats (dq 1.77 %, P<0.001) and sugars (dq 2.02 %, P<0.001), showed no association with the HEI. Differences in the intake of nutrients and foods across quintiles of consumption of bakery products, sweetened soft drinks and yogurt were usually very small. We conclude that the impact of the consumption of bakery products, sweetened soft drinks and yogurt on the quality of the diet of Spanish children is only modest, although it may contribute to aggravating certain unhealthy characteristics of their diet, particularly excess energy, saturated fats and sugars. Therefore, consumption of bakery products and sweetened soft drinks should be moderated, and priority given to consumption of low-fat, low-sugar yogurt. PMID- 12628037 TI - Role of dietary fat type in the development of adiposity from dietary obesity susceptible Sprague-Dawley rats. AB - The present study was designed to define how dietary fat type regulates body adiposity in dietary obesity-susceptible (DOS) Sprague-Dawley (SD) rats. Eighty three SD rats received a purified diet containing 50 g maize oil (MO)/kg for 3 weeks and then thirty-nine of the rats, designated as the DOS rats, were allotted to diets containing 160 g MO (DOS-MO), beef tallow (DOS-BT) or fish oil (DOS FO)/kg for 9 weeks. As a result of the experiment, the DOS-FO rats had significantly (P<0.05) reduced weight gain and abdominal and epididymal fat-pad mass than the DOS-MO and DOS-BT rats. Serum leptin level was also significantly (P<0.05) lower in the DOS-FO rats; however, hypothalamic leptin receptor (a and b) mRNA and neuropeptide Y expressions were not altered by dietary fat sources. A lower acetyl-CoA carboxylase mRNA expression in the liver was observed in the DOS FO group, whereas hepatic peroxisome proliferator-activated receptor-gamma mRNA and protein expressions were markedly elevated in the DOS-FO group compared with those in the other groups. We did not observe differences in acetyl-CoA carboxylase and peroxisome proliferator-activated receptor-gamma expressions in epididymal fat of the DOS rats consuming MO, BT or FO. It is concluded from our present observations that dietary fat type, especially that rich in FO, plays a potential role in down-regulation of adiposity by altering hepatic lipogenic genes, rather than feeding behaviour, in the DOS-SD rats. PMID- 12628038 TI - Renal relapse in Wegener's granulomatosis. PMID- 12628039 TI - Localized versus systemic vasculitis: diagnosis and management. AB - Localized vasculitis restricted to a specific anatomic site or organ is often histologically indistinguishable from more severe, systemic forms of vasculitis. By definition, localized vasculitis involves blood vessels within a confined vascular distribution or single organ without clinical evidence of generalized inflammation. Important factors that determine treatment and prognosis, in what appears initially to be a localized process, include histopathologic type, organ site, and the presence of systemic inflammatory markers and symptoms. The major issue is whether single organ vasculitis is actually an isolated form of the disease in which case surgical excision is curative, or whether the single organ involvement is simply a precursor of more threatening systemic vasculitis. The Birmingham Vasculitis Activity Score is a valuable tool to identify those patients with concurrent systemic involvement. The physician's recognition of a localized versus systemic vasculitic process is important in terms of making the correct diagnosis, prescribing treatment, and arranging appropriate clinical follow-up. PMID- 12628040 TI - Ocular vasculitis. AB - Systemic vasculitis can affect practically any structure in or around the eye. Although the ocular presentations of vasculitic disease are highly variable and may be quite subtle, it is important for physicians to understand some of the basic concepts, critical findings, and potential significance of ocular signs and symptoms. In addition to preserving sight, awareness of specific ocular syndromes may afford a critical clue to undiagnosed or newly active systemic vasculitis. A brief discussion of the most common ocular syndromes seen in systemic vascular disease is presented, with special attention given to the signs that may be appreciated by non-ophthalmologists. Recent concepts of ocular immunologic therapy are also reviewed, especially as they relate to the use of systemic immunomodulating drugs, because care of these patients often requires close cooperation between ophthalmologists, primary care physicians, and rheumatologists. PMID- 12628041 TI - Pulmonary-renal vasculitic disorders: differential diagnosis and management. AB - Pulmonary-renal syndrome (PRS) is a combination of diffuse pulmonary hemorrhage and glomerulonephritis. Pulmonary-renal syndrome is not a single entity and is caused by a variety of conditions, including Goodpastures syndrome associated with autoantibodies to the glomerular and alveolar basement membranes, various forms of primary systemic vasculitis associated with serum positivity for antineutrophil cytoplasmic antibodies (ANCA), cryoglobulinemia, systemic lupus erythematosus, systemic sclerosis, antiphospholipid syndrome, environmental factors, and drugs. The majority of cases of PRS are associated with ANCAs. The antigen target in Goodpastures syndrome is the alpha-3 chain of type IV collagen. The antigen target in PRS associated with systemic vasculitis is proteinase-3 and myeloperoxidase. Pulmonary-renal syndrome has been observed from the first to the ninth decade of life. The widespread adoption of serologic testing performed in an appropriate clinical context hopefully will limit diagnostic delay. The goals of treatment in PRS are to remove the circulating antibodies, to stop further production of autoantibodies, and to remove any antigen that stimulates antibody production. Treatment is based on plasmapheresis, steroids, and cyclophosphamide; however, infections are frequent contributors to death, and less toxic alternatives may improve outcome and prognosis resulting in a long-term survival. The degree of renal function and the percent of crescents on renal biopsy are better predictors of outcome. Renal transplantation can be safely carried out in PRS. PMID- 12628042 TI - Central nervous system vasculitis. AB - Vasculitis of the central nervous system (CNS) is classified as primary angiitis or as vasculitis secondary to a variety of diseases. A wide spectrum of clinical features may occur. A definite diagnosis is hampered by the difficulty in obtaining tissue for histology. Consequently, a diagnosis is frequently made on the basis of clinical presentation, brain magnetic resonance imaging, and cerebral angiography without pathologic confirmation. Recent experience shows that there are multiple other conditions that can mimic CNS vasculitis, many of which have different therapies. Most patients with CNS vasculitis should be treated aggressively with a combination of immunosuppressive medications. The prognosis is greatly improved with early recognition and therapy. PMID- 12628043 TI - Granulomatous vasculitis. AB - Granulomatous vasculitis is a subset of systemic necrotizing vasculitis and has granulomatous inflammation as the main histopathologic feature. Etiopathogenesis remains poorly understood, although recent advances suggest an important role for certain pro-inflammatory cytokines, such as tumor necrosis factor-alpha. They are a heterogeneous group of clinical disorders with protean manifestations. Serologic abnormalities are present, and the presence of granular cytoplasmic staining-antineutrophil cytoplasmic antibodies is most important and is particularly useful for the diagnosis of active Wegener's granulomatosis. Corticosteroids and cyclophosphamide remain very useful in the treatment of most of these disorders. PMID- 12628044 TI - Infectious etiology of vasculitis: diagnosis and management. AB - Vasculitides are a heterogeneous group of syndromes characterized by inflammation of the vessel wall. Several microbial pathogens have been known or suspected to cause vasculitis, and the development of molecular biology has promoted the search and confirmation of infectious agents in idiopathic vasculitis. Though several agents present primarily as an infectious process with vasculitis as an occasional manifestation, vasculitis may be the major manifestation of disease. Less definitive, and more controversial, is the role of infection and inflammation of the vessel wall in the pathogenesis of atherosclerotic disease. Clinical features can be nonspecific, and a high index of suspicion is required in order to make a diagnosis of vasculitis. Infection should always be excluded based on appropriate cultures and serologic assays, and, if confirmed, early and aggressive treatment should be instituted. However, in many instances, especially if there is a delay in the diagnosis, surgical intervention becomes necessary to treat the associated anatomic and physiologic sequelae. PMID- 12628045 TI - Endothelin-I receptor antagonist for the treatment of pulmonary arterial hypertension in systemic sclerosis. PMID- 12628046 TI - Mast cells in the pathogenesis of fibrosis. AB - It has been appreciated for quite some time that a "round cell" inflammatory infiltrate is present in the dermis during the early stages of scleroderma. Considerable research has emanated from this basic observation, with the assumption that interactions between immune cells and fibroblasts are paramount in the genesis of fibrosis. The large number of mast cells that accumulate early in involved tissues has been less appreciated in recent studies. The purpose of this update is to demonstrate how mast cells interact with fibroblasts in a manner that leads to fibroblast activation and subsequent extracellular fibrosis. Furthermore, the notion that myofibroblasts represent a critical fibroblast phenotype in sclerosing disorders, such as scleroderma, has also gained considerable support. The relationship of tissue mast cells to the generation of a myofibroblast phenotype has been the focus of several recent reports and will also be discussed. PMID- 12628047 TI - Microchimerism and scleroderma: an update. AB - Microchimerism has been defined by the presence of a low number of circulating cells transferred from one individual to another. This transfer takes place naturally during pregnancy, between mother and fetus, or between fetuses in multigestational pregnancies. Furthermore, the establishment of microchimerism can also occur during blood transfusion, as well as during bone marrow and solid organ transplants. Recently, microchimeric cells have been implicated in the pathogenesis of autoimmune diseases, particularly systemic sclerosis. Studies have demonstrated an increased presence of microchimeric cells in peripheral blood and tissues from patients with systemic sclerosis, and, more recently, microchimeric cells were demonstrated to be specifically activated and capable of recognizing patient human leukocyte antigens. PMID- 12628048 TI - Ethnicity and race and systemic sclerosis: how it affects susceptibility, severity, antibody genetics, and clinical manifestations. AB - Most studies have suggested that ethnic factors impact significantly on systemic sclerosis. Extensive epidemiologic studies have been carried out in white individuals, and limited data suggest that blacks are affected twice as frequently; Japanese patients have a lower prevalence than whites. This highest rate that has been described has been in Choctaw Native Americans. Blacks have a lower age at onset, as well as a higher frequency of diffuse skin involvement, pulmonary disease, and an overall worse prognosis than whites. Limited data in Hispanics and Native Americans suggest that they have more severe disease than whites. Whites have the highest frequency of anti-centromere antibodies (associated with limited skin involvement and less pulmonary fibrosis), whereas blacks have a higher frequency of anti-ribonucleoprotein and fibrillarin autoantibodies; the latter is a nucleolar antibody associated with a poorer prognosis. Ethnic differences are also seen for associations with non-major histocompatibility complex genes, such as FBN1 (fibrillin) genes, in Choctaws and Japanese and SPARC (osteonectin) in whites, Hispanics, and Choctaws. Although these facts do not entirely rule out socioeconomic factors associated with ethnicity, nevertheless ethnicity has an important impact on the pathogenesis of systemic sclerosis, perhaps because of genetic factors. PMID- 12628049 TI - Treatment of Raynaud's phenomenon: new insights and developments. AB - Although certain broad principles of treatment apply to all patients with Raynaud's phenomenon, the exact approach will vary depending on the nature and severity of the underlying digital vascular problem. Advances in the understanding of the pathophysiology of Raynaud's phenomenon are directing new lines of therapy. This review considers general (nonpharmacologic) measures, the different types of drug therapy, and the role of surgery. Recent advances in the use of more conventional treatments, such as calcium channel blockers, are discussed and also newer agents that are currently being researched, for example, endothelin-1 receptor antagonists and nitric oxide donors. The necessity of prompt assessment and treatment of acute digital ischemia, which is a medical emergency, is highlighted. PMID- 12628050 TI - The psychiatric management of end-of-life pain and associated psychiatric comorbidity. AB - Involvement by psychiatrists in the care of patients who are terminally ill has been limited historically; however, psychiatrists increasingly are participating in the care of these people who are facing the most challenging times of their lives. Pain management is considered to be an area of subspecialization for psychiatrists beyond their traditional role of being psychopharmacologists and psychotherapists. Biologically focused psychiatrists are able to address neuropsychiatric disorders, including pain and depression in the medically ill, and actively improve the quality of life for dying patients and their family members. This article provides a review of the recent literature that has addressed the involvement of psychiatrists in end-of-life care. Beginning with an example of the scientific aspects of cancer-related treatment from a patient's perspective, and into the major treatment considerations, this article addresses pain, its recognition, and management challenges when the end of life approaches. The prompt recognition of frequently overlooked and underestimated concomitant depressions, delirium, dementia, and other mental disorders is important for mental health specialists. The importance of psychiatric care for patients who are terminally ill and the role of psychiatrists in the phase of care also are discussed. PMID- 12628051 TI - The relevance of health anxiety to chronic pain: research findings and recommendations for assessment and treatment. AB - Health anxiety is a dimensional construct that is characterized by extreme concern about one's health at one end of the continuum and lack of concern about health on the other. Research evidence is consistent with the notion that high levels of health anxiety among patients with pain represent a poor prognostic indicator. Moreover, the experience of chronic pain can serve to increase health anxiety levels. This article concludes with a series of practical recommendations to assist practitioners in the assessment and management of patients with health anxiety. PMID- 12628052 TI - Pain: psychiatric aspects of impairment and disability. AB - Patients with chronic pain frequently experience a complex and convoluted journey through the health care system that often is unrewarding for all involved. As job satisfaction and financial security diminish during our economic recession, the impact of the disability epidemic becomes more profound. There often is no direct correlation between objective impairment and a patient's request for disability status. Injured workers maintained on workers' compensation may have an increased risk for developing chronic pain syndromes unresponsive to conventional treatments. These patients may have significant financial, psychosocial, and environmental reinforcement for maintenance of their disability and little incentive to return to work. Excessive pain behavior may lead to unnecessary diagnostic testing or invasive procedures and result in iatrogenic complications and prolonged disability. Patients with chronic pain syndrome who have not had psychosocial treatment may not be at maximum medical improvement. The purpose of this paper is to help the readership identify types of psychiatric, psychologic, and psychosocial issues that can coexist in patients with chronic pain who are applying for disability, and to help treating physicians avoid contributing to iatrogenic pain and disability by performing needless and potentially harmful procedures on patients who may be better served with an emphasis on psychiatric or psychologic care. PMID- 12628053 TI - Cognitive impairment in patients with chronic pain: the significance of stress. AB - This review article examines the role of emotional distress and other aspects of suffering in the cognitive impairment that often is apparent in patients with chronic pain. Research suggests that pain-related negative emotions and stress potentially impact cognitive functioning independent of the effects of pain intensity. The anterior cingulate cortex is likely an integral component of the neural system that mediates the impact of pain-related distress on cognitive functions, such as the allocation of attentional resources. A maladaptive physiologic stress response is another plausible cause of cognitive impairment in patients with chronic pain, but a direct role for dysregulation of the hypothalamic-pituitary-adrenocortical axis has not been systematically investigated. PMID- 12628054 TI - Chronic nonmalignant pain and violent behavior. AB - Research suggests that violence has entered the medical setting to a remarkable degree, causing medical professionals to be at the highest risk for becoming the victims of assaults and violent acts. This article reviews general theories of aggression and research on these theories, and uses them to assess risk factors in patients with chronic pain. There are data to suggest that pain may increase the risk of aggressiveness in some patients. However, it may decrease the risk in others paradoxically. The research available underscores the need for evaluating patients with pain for the risk of violent or aggressive behavior; specific recommendations are made in this regard. PMID- 12628055 TI - Topiramate in the treatment of cluster headache. AB - Cluster headache is a well-characterized, strictly unilateral headache with cranial autonomic features and can be classified as episodic or chronic. Cluster attacks reliably are short-lived, often have a clockwise regularity, and can occur daily for weeks or months during an active cluster period. Pharmacologic treatment for this disorder can be divided into abortive and prophylactic agents. Prophylactic agents aim to quickly induce and maintain a remission. Short-term prophylaxis may be attained with the use of steroids, ergotamine, or methysergide, but these agents are not as suitable for continuous use. Verapamil and lithium commonly are used for longer periods and other agents, such as melatonin and baclofen, also are considered useful. There has been increased interest in the use of anticonvulsants for pain syndromes such as primary headache disorders. This includes topiramate use for cluster prophylaxis; a number of open-label studies have had encouraging results. This article provides an overview of topiramate and the open-label studies of this agent in the prevention of cluster headache. PMID- 12628056 TI - Topical agents in the treatment of cluster headache. AB - This article discusses topical intranasal medications in the treatment of cluster headache. PMID- 12628057 TI - Epidemiology of cluster headache. AB - Cluster headache is rare, occurring in less than 1% of the population. Studies suggest that, in addition to the pain and associated autonomic disturbances recognized to be characteristic of the syndrome, patients also may experience nausea, photophobia, behavioral agitation, or restlessness. A decreasing male:female ratio also has been noted, perhaps attributable to lifestyle trends adopted by more women that were previously associated with men, such as tobacco use, alcohol consumption, and working outside of the home. The relationship between cluster headache and hormonal events does not appear to be strong. Hormonal influences on the chronic form of cluster headache in women are a subject of investigation. The emerging understanding of the genetics of cluster headache increasingly suggests a genetic component, with familial transmission now recognized to be more common than previously appreciated. Head trauma, coronary artery disease, and migraine appear to be present in more patients with cluster headache than can be explained by chance alone. Ethnic and racial differences in prevalence are less well understood. PMID- 12628058 TI - Cluster headaches and sleep disorders. AB - Cluster headaches are characterized by unilateral paroxysmal attacks of severe pain with associated symptoms. The headaches occur during particular sleep stages and are associated with other chronobiologic factors. Several sleep disorders have been associated with the occurrence of cluster headache; multiple hormonal influences affect the relationship between sleep and headache. Melatonin and other treatments that affect circadian rhythm have been suggested for the treatment of cluster headache. Obstructive sleep apnea can occur in patients with cluster headache; attempts to treat one disorder may influence the other. Sleep disorders such as insomnia and narcolepsy also may be associated with and influence cluster headaches. This article examines the relationship between the various sleep disorders and cluster headache, and reviews current research. Normal and abnormal sleep and details of treatments for specific sleep disorders that may decrease the frequency and severity of cluster headaches also are discussed. The relationship between obstructive sleep apnea, which is the most common sleep disorder, and cluster headache is discussed in detail. PMID- 12628061 TI - Antiepileptic Drugs in Children with Developmental Delays and Behavioral Problems. AB - Children with multiple handicaps, including developmental delays and behavioral problems, frequently have epilepsy that requires treatment. Different strategies should be used for acute treatment versus chronic treatment. Experience suggests that there is no true drug of choice that can be used for all children with epilepsy; however, seizure type, epilepsy syndrome, and likely efficacy and side effect spectrum of the antiepileptic drug (AED) will determine the best treatment for each child. In general, phenobarbital has posed multiple behavioral problems, and, thus, is less commonly used. Carbamazepine is the most frequently used drug for the treatment of partial seizures with or without secondary generalization, and valproate is the long established, broad spectrum AED for children with generalized convulsive and nonconvulsive seizures. Newer AEDs allow alternative treatments to those with adverse effects to the more frequently used medications and may have cognitive or behavioral advantages. PMID- 12628059 TI - Diagnosis and Treatment of Nonconvulsive Status Epilepticus in an Intensive Care Unit Setting. AB - Nonconvulsive status epilepticus (NCSE) in adults is a heterogeneous epileptic emergency and includes absence status (AS), complex-partial status epilepticus (CPSE), and the status epilepticus of epileptic encephalopathy (SEEE). The latter seems to be strikingly frequent among patients in intensive care units (ICU). Diagnosis of NCSE is difficult, but has to be made quickly. It relies on clinical signs and a confirmation electroencephalography (EEG). According to the different etiologies and outcomes of AS, CPSE, and SEEE, treatment has to be individually adapted, but needs to follow some basic principles--treatment should take place in the ICU and be monitored by continuous EEG. With a few exceptions, the first drug is an intravenous benzodiazepine, mainly lorazepam. Intravenous fosphenytoin or phenytoin or valproate may follow next. If some forms of NCSE are resistant to first- and second-line treatments, single or combinations of anesthetics and enteral antiepileptic drugs (AEDs) may be added. This opinion is not evidence based, and randomized controlled prospective trials to evaluate optimal treatment of NCSE are of first priority. PMID- 12628060 TI - Vagus Nerve Stimulation As Treatment for Epileptic Seizures. AB - Vagus nerve stimulation is a unique therapy for epileptic seizures. Two randomized controlled trials in patients with medically refractory partial seizures have demonstrated efficacy, leading to US Food and Drug Administration approval of vagus nerve stimulation therapy in 1997. Extensive safety testing has not revealed significant effects on cardiac, respiratory, or gastrointestinal function, though recent reports of intraoperative asystole and sleep-related airway obstruction have raised concerns. Vagus nerve stimulation is indicated for adjunctive therapy of partial-onset seizures in children and individuals older than 12 years (Class I evidence). Based on controlled, randomized trials, approximately 30% of these patients can be expected to have at least a 50% decrease in overall seizure frequency. Vagus nerve stimulation efficacy is similar to that of several newer antiepileptic drugs when used in similar populations in controlled, randomized trials. Long-term follow-up studies suggest continued efficacy over more than 1 year (Class III evidence). Case series suggest similar or greater efficacy in younger children, and in patients with refractory generalized seizures, including those associated with the Lennox Gastaut syndrome (Class III evidence). Vagus nerve stimulation is appropriate therapy for patients with medically refractory epileptic seizures who are not optimal candidates for resective epilepsy surgery. PMID- 12628064 TI - Corticobasal Degeneration. AB - Corticobasal degeneration is one of the neurodegenerative tauopathies, which are defined as a group of heterogeneous dementias and movement disorders that are characterized neuropathologically by prominent intracellular accumulations of abnormal filaments formed by the microtubule-associated protein tau. Although there are no curative treatments, symptomatic and supportive management can be helpful. Many new therapies are still under development. However, more needs to be learned about the pathogenesis and molecular biology of this disease before an effective therapy can be developed. PMID- 12628065 TI - Psychogenic Movement Disorders. AB - Psychogenic movement disorders are one of the most challenging conditions encountered by clinicians. Patients with these disorders represent the overlap of psychiatry and neurology and need care from these specialties. Issues within the patient's family, questions about secondary gain, and prejudices among other clinicians often complicate evaluation and treatment. Diagnosis is further complicated by lack of laboratory and other tests to reliably diagnose movement disorders; most movement disorders are diagnosed based on history and clinical phenomena. Treatment is best conducted within the context of a multidisciplinary team experienced in working with these patients and with one another. Identification and treatment of underlying psychopathology, such as major depression or anxiety disorders, is also of great importance. PMID- 12628062 TI - Parkinson's Disease: Surgical Options. AB - Surgical therapy for Parkinson's disease (PD) has been a treatment option for over 100 years. Advances in the knowledge of basal ganglia physiology and in techniques of stereotactic neurosurgery and neuroimaging have allowed more accurate placement of lesions or "brain pacemakers" in the sensorimotor regions of target nuclei. This, in turn, has led to improved efficacy with fewer complications than in the past. Currently, bilateral deep brain stimulation (DBS) of the subthalamic nucleus (STN) or the internal segment of the globus pallidus (GPi) is the preferred option (and is approved by the US Food and Drug Administration) for the surgical treatment of PD. The most important predictors for outcome for DBS for PD are patient selection and electrode location. Patients should have a documented preoperative improvement from dopaminergic medication of at least 30% in the patient's Unified Parkinson's Disease Rating Scale motor disability scores. A levodopa challenge may be needed to document the best "on" state. Dementia or active cognitive decline must be excluded. Active psychiatric disease should be treated preoperatively. Patients should be motivated, with good support systems, and committed to the postoperative management of DBS therapy. Deep brain stimulation should be considered when the patient begins to experience dyskinesia and on-off fluctuations despite optimal medical therapy. Deep brain stimulation is not a good option at the final stages of the disease because of the increased incidence of dementia and severe comorbidity. The DBS electrode should be placed in the sensorimotor region of the GPi or STN. Subthalamic nucleus and GPi DBS can improve all motor aspects of PD, as well as predictable "on" time, without dyskinesia or fluctuations. On average, STN DBS results in a greater reduction of dopaminergic medication compared with GPi DBS. Because of the smaller size of the target region, the pulse generator battery life is longer with STN then with GPi DBS. Deep brain stimulation programming is a skill that is readily learned and may be required of all neurologists in the future. Emerging surgical therapies are restorative, and they aim to replace or regenerate degenerating dopaminergic neurons. These include embryonic mesencephalic tissue transplantation, human embryonic stem cell transplantation, and gene-derived methods of intracerebral implantation of growth factors and dopamine- producing cell lines. It will be important to determine whether DBS, if performed before the onset of motor response complications to medical therapy, may prevent this stage of disease altogether or delay it for a significant period of time. The same question applies to the future with restorative therapy. PMID- 12628063 TI - Autonomic Nervous System Dysfunction in Parkinson's Disease. AB - Autonomic nervous system (ANS) dysfunction is common in Parkinson's disease (PD), affects 70% to 80% of patients, and causes significant morbidity and discomfort. Autonomic nervous system dysfunction symptoms in PD include sexual dysfunction, swallowing and gastrointestinal disorders, bowel and bladder abnormalities, sleep disturbances, and derangements of cardiovascular regulation, particularly, orthostatic hypotension. Autonomic nervous system dysfunction in PD may be caused by an underlying degenerative process that affects the autonomic ganglia, brainstem nuclei, and hypothalamic nuclei. Anti-parkinsonian medications can cause or worsen symptoms of ANS dysfunction. The care of a PD patient with ANS dysfunction relies on its recognition and directed treatment, including coordinated care between the neurologist and appropriate subspecialist. Pharmacotherapy may be useful to treat orthostasis, gastrointestinal, urinary, and sexual dysfunction. PMID- 12628066 TI - Gait and Balance Dysfunction in Adults. AB - Determining the precise cause of gait dysfunction in adults is often difficult because of the multifactorial nature of the disorder. Additionally, elderly patients have other comorbidities that further complicate their diagnosis. A proper history and physical examination, however, often allow the clinician to arrive at the correct diagnosis. Once a diagnosis is reached, appropriate therapeutic decisions can be made. Patients presenting with Parkinsonism need a thorough evaluation to rule out potentially reversible conditions, such as normal pressure hydrocephalus. Patients with idiopathic Parkinson's disease usually develop gait difficulty and freezing episodes late in the course of the illness. Another important cause of gait disturbance in adults is the cerebellar ataxias. Among the sporadic forms, gluten sensitivity is an important consideration. Identification of this entity is important, because the disease process can be halted with a gluten-free diet. Another group is the paraneoplastic ataxias, which can often be diagnosed in the proper clinical setting. Most of the adult onset hereditary ataxias are autosomal dominant conditions. Except for the episodic ataxias, treatment of these conditions has been disappointing. Mixed results have been obtained with the use of amantadine, buspirone, and 5 hydroxytryptophan. Physical therapy plays an important role in the gait rehabilitation of these patients. Over the past several years, researchers have developed a greater understanding of motor control and how it relates to freezing. Clinicians can now train patients to use external cues to overcome their motor blocks. Another important advance has been the development of subthalamic nucleus deep brain stimulation in the treatment of patients with troublesome peak dose dyskinesia and other motor fluctuations. Subthalamic nucleus deep brain stimulation should be considered when best medical treatment fails. Cortical myoclonus can be treated with levetiracetam, which has US Food and Drug Administration approval as an antiepileptic agent. It has been quite effective in the treatment of myoclonus and should be considered when other medications fail. PMID- 12628067 TI - Endoscopic Interventions in the Biliary Tract. AB - In patients with common bile duct stones, the endoscopic removal of such stones has become the standard therapy. The surgical approach is indicated only for stones that cannot be extracted endoscopically. In biliary obstruction due to pancreaticobiliary malignancy, endoscopic interventions represent the first-line therapy in the palliative situation. If endoscopic access is not possible, the percutanous approach often represents an alternative, followed by palliative surgical options. Biliary strictures or bile leaks after liver transplantation very often may be treated effectively by endoscopic dilatation or temporary stenting. In most cases, endoscopic therapy of such problems represents the first option before the percutaneous or surgical approach is indicated. In primary sclerosing cholangitis, dominant biliary strictures develop frequently and endoscopic treatment allows their opening in most cases. There is no real alternative to endoscopy in this situation. The situation is more complex in postoperative biliary strictures and chronic pancreatitis. In these conditions, advantages and disadvantages of the endoscopic versus the surgical approach have to be evaluated to find the most effective form of treatment in the individual situation. The surgical intervention often represents the better alternative. PMID- 12628069 TI - Diagnosis, Staging, and Treatment of Cholangiocarcinoma. AB - Attention has to be focused on earlier detection of cholangiocarcinoma, especially in high-risk patients such as those with primary sclerosing cholangitis (PSC). Enhanced techniques for diagnosing this disease include imaging studies such as positron emission tomography (PET) scanning, and cytologic analysis including digital image analysis (DIA) and fluorescent in situ hybridization (FISH). Magnetic resonance imaging and endoscopic ultrasonography have also improved and simplified preoperative staging. Despite recent advances in the endoscopic management of cholangiocarcinoma, this neoplastic disease is still fatal in the majority of patients. Surgical resection or liver transplantation offers the only possibility for curing this disease in its early stages. However, most patients present with advanced disease that is not amenable to such therapy. For those patients presenting with unresectable carcinoma above the cystic duct without intrahepatic or extrahepatic metastases, orthotopic liver transplantation combined with preoperative irradiation and chemotherapy is available and demonstrates improved survival on the basis of a recent study conducted at the Mayo Clinic. In the future, chemopreventive strategies aimed at blocking the links between inflammation (eg, nitric oxide synthase and cylcooxygenase 2 inhibitors) and carcinogenesis may help prevent this often fatal disease in high-risk patients (eg, patients with PSC). PMID- 12628070 TI - Liver Transplantation for Cholestatic Liver Disease. AB - Liver transplantation is an effective form of therapy for patients with end-stage cholestatic disease that improves both survival and quality of life. Liver transplantation is very effective for the treatment of intractable pruritus but less effective for the treatment of lethargy. Survival rates are good (more than 70% at 5 years); these patients are at greater risk of developing acute and chronic rejection and are more likely to require long-term immunosuppression. Primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC) recur in the graft. Recurrent PSC may be difficult to differentiate from secondary sclerosing cholangitis, but it recurs in up to 60% of patients at 5 years and may reduce graft survival. PBC recurrence, noted in up to 40% of patients at 10 years, has little effect on graft survival with respect to cancers. Patients with PSC are at greater risk of both colonic cancer (which may be reduced by ursodeoxycholic acid) and cholangiocarcinoma. Diagnosis of cholangiocarcinoma before transplantation usually contraindicates transplantation. The main challenges facing liver transplantation are the need to expand the donor pool and the need to find immunosuppressive regimens with fewer long-term toxicities. PMID- 12628068 TI - Treatment Options for Primary Biliary Cirrhosis and Primary Sclerosing Cholangitis. AB - Primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC) are chronic cholestatic liver diseases that affect 0.5 to 40 per 100,000 and 1 to 6 per 100,000 Americans, respectively. Prompt recognition and management of the clinical manifestations of these diseases is essential for the patients' well being and ultimate outcome. Ursodeoxycholic acid (UDCA), 13 to 15 mg/kg per day, is the standard therapy for PBC and should be offered to every patient. It has been shown to slow progression of the disease and prevent the need for liver transplantation, which is the last recourse for patients with end-stage disease. However, there is no effective therapy for PSC yet. Patients are managed symptomatically, with surgical or endoscopic interventions as needed in cases of significant biliary obstruction. Complications of chronic cholestasis are seen in both PBC and PSC, with pruritus and fatigue being the most common complaints. The first choice for the treatment of pruritus is still cholestyramine, starting at 4 g/d. The pathogenesis of fatigue is poorly understood in this population; unrecognized hypothyroidism should be excluded. The use of antidepressants is currently under evaluation, but there is no specific therapy for fatigue as of yet. For prevention of severe osteoporosis, we recommend supplementation with 800 IU vitamin D and 1500 mg calcium/d. In patients with PBC and established osteoporosis, the use of alendronate and vitamin K appears to cause an increase in bone mineral density. Further studies are necessary before either of these drugs is routinely recommended. Finally, fat-soluble vitamin deficiencies are noted with more advanced disease. We recommend that serum levels be checked in high-risk patients, and that vitamins are replaced as appropriate with water soluble supplements. However, other causes of malabsorption must be ruled out, including pancreatic insufficiency and celiac sprue. PMID- 12628071 TI - Intrahepatic Cholestasis of Pregnancy. AB - Intrahepatic cholestasis of pregnancy (or obstetric cholestasis) is a liver disorder that occurs in late pregnancy. Despite the potential adverse maternal and fetal/neonatal outcomes, cholestasis of pregnancy is often neglected and treated expectantly. More research is needed to improve the molecular and genetic understanding of the disease and to define a safe and effective medical treatment that improves clinical outcome. Ursodeoxycholic acid is considered to be a safe treatment option in the third trimester, but further randomized controlled trials are needed before ursodeoxycholic acid treatment can be generally recommended. Ursodeoxycholic acid is preferentially administered to patients with severe cholestasis (onset before week 33 or serum bile acid levels > 70 mmol/L) or to patients with a history of sudden fetal death, while maintaining close obstetric and regular biochemical surveillance (transaminases, bilirubin, and bile acid levels). Ursodeoxycholic acid can decrease pruritus and ameliorate liver tests, but effects on obstetric complications are ambiguous. S-Adenosylmethionine, dexamethasone, and cholestyramine can provide some relief of itching. Because none of these drugs have been shown to be harmful to mother or fetus, the individual woman and her clinician may decide whether to try one of the treatments described. PMID- 12628072 TI - Stress-related Mucosal Disease. AB - Stress-related mucosal disease (SRMD) includes stress-related injury (superficial mucosal damage) and stress ulcers (focal deep mucosal damage). Both types are caused by mucosal ischemia, and both show a propensity for the acid-producing corpus and fundus. Prophylaxis of stress ulcers may reduce major bleeding but, so far, has not been shown to improve survival. The most widely used drugs for stress-related injury are the intravenous histamine H(2)-receptor antagonists. Proton pump inhibitors (PPIs) are the most potent acid-suppressive pharmacologic agents. The available PPIs significantly increase gastric pH for up to 24 hours after one dose. Tolerance does not develop, and adverse effects are few. Preliminary studies have demonstrated a significant reduction in SRMD bleeding for patients receiving PPI prophylaxis. PPIs may become an effective tool for reducing the incidence of SRMD in critically ill patients. PMID- 12628073 TI - Treatment of Helicobacter pylori Eradication Failures. AB - Even with the current most effective treatment regimens, about 10% to 20% of patients will fail to eradicate Helicobacter pylori infection. Therefore, in designing a treatment strategy, we should not focus on the results of primary therapy alone but also on the final (overall) eradication rate. The choice of a second-line treatment depends on which treatment was used initially, because retreatment with the same regimen is not recommended. Therefore, it seems that performing culture after a first eradication failure is not necessary and assessing H. pylori sensitivity to antibiotics only after failure of the second treatment is suggested in clinical practice. Different possibilities of empirical treatment are suggested. After failure of proton pump inhibitor (PPI)-amoxicillin clarithromycin, quadruple therapy has been generally used. More recently, replacing the PPI and the bismuth compound by ranitidine bismuth citrate has also achieved good results. After PPI-amoxicillin-nitroimidazole failure, retreatment with PPI-amoxicillin-clarithromycin has proved to be effective. Finally, the first therapy should not combine clarithromycin and metronidazole in the same regimen because of the problem of resistance against both antibiotics. Recently, rifabutin-based rescue therapies have been shown to constitute an encouraging strategy for eradication failures because they are effective for H. pylori strains resistant to antibiotics. PMID- 12628074 TI - Pharmacologic Treatment of Upper Gastrointestinal Bleeding. AB - The first goal of therapy is to assess hemodynamic stability in a patient presenting with evidence of upper gastrointestinal bleeding and, second, to maintain hemodynamic stability using crystalloids or packed red blood cells. The diagnosis of the cause of upper gastrointestinal bleeding should be performed using endoscopic techniques, which should be performed early. Therapy directed at treating the cause of upper gastrointestinal bleeding should be initiated at the time of endoscopy. Pharmacologic management should be based on the prevention of ulcer rebleeding and should be initiated at the time of endoscopic diagnosis. Surgery should be considered only in cases when endoscopic and pharmacologic treatments are deemed a failure. PMID- 12628075 TI - Zollinger-Ellison Syndrome. AB - Zollinger-Ellison syndrome (ZES) is caused by a gastrin-producing tumor called a gastrinoma, which results in gastric acid hypersecretion. Gastrin stimulates the parietal cell to secrete acid directly and indirectly by releasing histamine from enterochromaffin-like (ECL) cells, and induces hyperplasia of parietal and ECL cells. ZES should be suspected in patients with severe erosive or ulcerative esophagitis, multiple peptic ulcers, peptic ulcers in unusual locations, refractory peptic ulcers, complicated peptic ulcers, peptic ulcers associated with diarrhea, and a family history of multiple endocrine neoplasia type 1 (MEN 1) or any of the endocrinopathies associated with MEN-1. The initial diagnostic test for ZES should be a fasting serum gastrin level when antisecretory medications are discontinued. If the gastrin level is elevated, gastric acidity should be assessed through pH or gastric analysis. It should be noted that hypochlorhydria causes feedback stimulation of antral gastrin secretion. In suspected cases of ZES with mild hypergastrinemia, the secretin stimulation test may be useful. Initial treatment for ZES should be oral high-dose proton pump inhibitors. If parenteral therapy is needed, intermittent bolus injection of pantoprazole is recommended. Total gastrectomy and antisecretory surgery is rarely required. Somatostatin receptor scintigraphy (SRS) is the initial localization study of choice. Endoscopic ultrasound (EUS) may have a similar sensitivity for identifying primary tumors. A combination of SRS and EUS detects greater than 90% of gastrinomas. In patients without metastasis and without MEN 1, surgical cure is possible in 30%. It has been suggested that patients with gastrinomas larger than 2.5 cm, irrespective of whether they have MEN-1, should undergo surgical resection in an effort to decrease the risk for metastasis. PMID- 12628077 TI - Conflict of interest in health care: a subtle but significant issue in women's health. PMID- 12628078 TI - Evaluation and treatment of lipid disorders in women. AB - The latest report from the National Cholesterol Education Program has reaffirmed that the primary lipid goal for the prevention of atherosclerotic vascular disease (AVD) is to achieve a normal low-density lipoprotein (LDL) cholesterol (<130 mg/dL) by diet in normal individuals, and by diet and statin therapy in patients with multiple risk factors. Patients with any clinical AVD (which includes diabetes) will need a statin to achieve an optimal LDL cholesterol (<100 mg/dL). The recent Heart Protection Study might revise our thinking further. Patients at high risk achieved a reduction in mortality and vascular events taking simvastatin 40 mg, even if they had a low baseline LDL value. Individuals with the metabolic syndrome and insulin resistance do not typically have a very high LDL, but rather have elevated triglycerides and a low high-density lipoprotein (HDL) cholesterol. They, too, need to be treated with a statin, first to achieve an appropriate LDL goal. This is insufficient if the triglyceride value exceeds 200 mg/dL. They should be treated to achieve a non-HDL cholesterol goal (equal to total cholesterol minus HDL cholesterol) that is 30 mg/dL higher than the LDL goal. PMID- 12628076 TI - Treatment of Variceal Bleeding. AB - Mortality due to variceal bleeding secondary to portal hypertension has decreased significantly in the past 2 decades. Endoscopic therapy has been the mainstay of treatment for acute variceal bleeding. Variceal banding ligation has superceded injection sclerotherapy as the most popular treatment modality for acute bleeding. Multiple banding ligators are widely used with high success in restoring hemostasis. The combination of banding and sclerotherapy may be useful in preventing the early recurrence of varices and rebleeding after initial obliteration of varices. Selective vasoactive agents such as somatostatin analogs also improve the outcome of patients. Radiologic shunting has proven to be an effective salvage procedure when endoscopic treatments fail and may be a good intermediate-stage therapy while the patient is waiting for liver transplantation. PMID- 12628079 TI - Heart failure in women. AB - Congestive heart failure represents a growing health issue with significant morbidity, expense, and mortality. Unfortunately, despite heart failure affecting men and women equally, women historically have represented a minority in heart failure trials. Despite this disparity, treatment decisions rely heavily on these trials. Women with heart failure often have different clinical features than men, such as age of onset and comorbidities. Compared with men, women also demonstrate differences in remodeling and the response to injury, such as volume or pressure overload and myocardial infarction. We are only beginning to understand the clinical implications of these gender differences and their impact on pharmacologic treatments. After discussing these differences, a review of the agents useful in systolic failure is made, including angiotensin-converting enzyme inhibitors, b-blockers, digoxin, and aldosterone inhibition. Treatment of diastolic heart failure with empiric guidelines follows. PMID- 12628080 TI - Cardiovascular consequences of obesity. AB - Obesity is a problem of epidemic proportions that is associated with adverse cardiovascular outcomes. The long-term efficacy of currently available treatments is limited, and therapeutic benefits might paradoxically be achieved only at the expense of other serious adverse cardiovascular consequences. Owing to these limitations, new paradigms in the management of obesity are being explored, including expanded surgical and pharmacologic therapies. Recent discoveries in the pathogenesis of obesity, including the discovery of leptin, might lead to new treatment options. Leptin dysregulation within the central nervous system or in peripheral adipose tissue has a putative role in the pathogenesis of obesity. Furthermore, important differences exist between men and women in regard to leptin levels; these discrepancies might ultimately impact the therapeutic approach in a gender-specific manner. The goals of obesity therapy and the appropriate therapeutic endpoints are being evaluated to maximize the efficacy of therapy and improve cardiovascular outcomes, while diminishing the risk of adverse cardiovascular complications. PMID- 12628081 TI - Anorexigen-induced cardiac valvulopathy and female gender. AB - Obesity is a chronic disorder that is associated with significant co-morbidity and early mortality. Since the 1970s, amphetamines and amphetamine analogs have been trialed for weight loss. In 1997, the anorexigen agents, fenfluramine (Pondimin; AH Robins, Richmond, VA) and dexfenfluramine (Redux; Wyeth-Ayerst, Philadelphia, PA) gained worldwide attention when reports of associated cardiac valvulopathy and pulmonary hypertension emerged. A landmark report from the Mayo Clinic describing valvular heart disease (VHD) among women exposed to the anorexigen agent combination phentermine-fenfluramine ignited widespread concern, and the products were voluntarily withdrawn from the market by the manufacturers. Currently, the causal relationship between anorexigen agent use and valvulopathy has been validated, yet the extent and complexity of this important clinical issue remains ill-defined. PMID- 12628082 TI - Long QT syndrome: a preventable cause of sudden death in women. AB - Long QT syndrome is a hereditary disorder of cardiac ion channels causing abnormal electrical activation of the heart and leading to life-threatening ventricular tachycardia. Syncope, ventricular tachycardia, or sudden cardiac death in the absence of structural heart disease is the typical presentation, often starting in childhood. Women are more commonly affected by both the congenital form and the drug-induced acquired form of this disease and are particularly at risk in the postpartum period. The long QT syndrome can be recognized by a prolonged QT interval on the electrocardiogram. Several mutations in six genes encoding subunits of cardiac potassium and sodium channels have so far been identified, and gene-specific epidemiology, risk stratification, and management are emerging. beta-blockers are the mainstay of therapy for primary prevention of cardiac events and implantable defibrillators for secondary prevention. Patient education, screening of family members, and increasing awareness of this disorder among physicians are important steps toward prevention of sudden death in these otherwise healthy young individuals. PMID- 12628083 TI - Treatment of arrhythmias during pregnancy. AB - During pregnancy, significant changes occur in the hormonal and hemodynamic state of women that make arrhythmias more likely to occur. Palpitations are frequently reported, but are usually found to be associated with sinus tachycardia. The incidence of paroxysmal supraventricular tachycardia is increased during pregnancy, whereas atrial fibrillation and ventricular tachycardia are rarely seen. Women with long QT syndrome experience significantly more cardiac events in the postpartum period, making beta-blocker therapy most important during this time. Acute treatment of arrhythmias for pregnant women is much the same as that for nonpregnant patients. However, chronic drug therapy during pregnancy should be reserved for only the frequent, hemodynamically significant arrhythmia episodes. PMID- 12628084 TI - Myocardial infarction and unstable angina: gender differences in therapy and outcomes. AB - In the United States, coronary heart disease (CHD) is the leading cause of death for women. Combined with hypertension, stroke, and other vascular conditions, one in every two women dies annually owing to a cardiovascular condition. During the past 20 years, overall death rate from CHD has declined, but the rate of decline has been lower for women compared with men. Trials and studies have demonstrated that CHD might be preventable; however, optimal treatment strategies for women are still in question. It is, therefore, essential that the medical profession continues its current efforts to focus on the development of clinical practice guidelines that critically evaluate both diagnostic and therapeutic treatment options for women with CHD. PMID- 12628085 TI - Ductal lavage in the screening of high-risk women. AB - Due to the high incidence of breast cancer and the excessively high false positive rate of current screening technologies, adjunctive risk assessment measures are needed. Nipple aspiration cytologic analysis and random fine-needle aspirations have been proposed; however, ductal lavage and cytologic analysis of the effluent is a more sensitive and specific test. The dilemma lies in what should be done subsequent to a ductal lavage yielding malignant or atypical cells. Discussion of the proceeding procedures and potential management strategies are discussed. PMID- 12628086 TI - Ultrasound in the management of breast disease. AB - Since its introduction in the 1960s, ultrasound (US) has never been more important in breast disease management as it is today. Its historical role as an adjunct modality to mammography in differentiating cystic from solid lesions has been widely expanded. US-guided sampling procedures represent widely accepted modalities in lesion evaluation. Preoperative localization under US guidance is one of the methods of choice for nonpalpable solid lesions. Screening breast US is currently being investigated and might prove useful for high-risk women and women with dense breasts. Continuous technologic advances, such as high-frequency transducers, new Doppler-based techniques, microbubble contrast agents, harmonic and compound imaging, three-dimensional US, elasticity imaging, US detection of microcalcifications, perfusion US imaging, and computer-aided diagnosis all carry promise of further increases in US utility in the diagnosis and detection of breast disease. PMID- 12628088 TI - [Vaccination schedule 2003]. PMID- 12628087 TI - Hormone replacement therapy in breast cancer patients and survivors. AB - Menopause is arguably the most important phase of a woman's social, physiologic, and personal life. Approximately 1.3 million women reach this age in the United States annually. In the past decade, numerous studies have correlated breast cancer and the use of ERT (estrogen replacement therapy) or HRT (hormone replacement therapy) in menopausal women. Whether this is an actual increase in the creation of new cancers or a result of a diagnostic or other bias has yet to be determined. Even more uncertainty surrounds the use of hormones once breast cancer is diagnosed. Previously, once a woman was diagnosed with an estrogen dependent tumor, ERT and HRT were simply forbidden. As discussed herein, that is no longer the case. PMID- 12628089 TI - [Breastfeeding in the "global strategy for infant and young child feeding"]. PMID- 12628090 TI - [Takayasu's arteritis]. AB - BACKGROUND: Takayasu's arteritis is a rare vasculitis in the pediatric population that affects the aorta and its branches. There are few studies with an appropriate number of patients and follow-up. OBJECTIVE: To describe the clinical manifestations, laboratory alterations, radiological findings, and treatment in eight children and adolescents with Takayasu's arteritis. METHODS: A retrospective analysis of patients' records from 1990 to 2001 was performed. RESULTS: There were six girls and two boys. The mean age at disease onset was five years and five months. The most common clinical manifestations were systemic findings and cardiovascular, dermatological and neurological abnormalities. In all patients inflammatory activity was high and in three patients the Mantoux test was strongly positive. The most common radiological findings were type IV and V. Treatment included steroids, methotrexate, cyclophosphamide, intravenous gamma globulin, and vascular surgery. Three patients presented sequelae. CONCLUSIONS: Takayasu's arteritis produces considerable morbidity and mortality. To make an early diagnosis, pediatricians should be aware of inflammatory systemic manifestations and cardiovascular abnormalities. To gain further knowledge of this entity prospective and ideally multicenter studies are required. PMID- 12628091 TI - [Continuous glucose monitoring with type 1 diabetes mellitus]. AB - BACKGROUND: Appropriate metabolic control of children with type 1 diabetes mellitus (DM) is based on frequent measurements of capillary glycemia. However, this method offers only partial information on fluctuations in glycemia during the day, while episodes of postprandial hyperglycemia and hypoglycemia, mainly nocturnal, go unnoticed. OBJECTIVES: To analyze pre- and postprandial blood glucose levels, as well as the presence and duration of hypoglycemic episodes in diabetic children aged more than 8 years old with more than one year of disease duration. METHODS: Seventeen patients of both sexes (mean age: 12 years old) with type 1 DM were monitored with the continuous glucose monitoring system (CGMS) during working days. Maximum values of pre- and postprandial glucose (1-3 hours after breakfast, lunch and dinner) were registered. Data were downloaded with a Com-station. RESULTS: The mean duration of sensor-wearing was 2.97 days. Pre- and postprandial values were high: mean preprandial values were between 144.9 and 160.5 mg % and mean postprandial values were between 230.4 and 248.8 mg %. The mean number of hypoglycemic episodes detected with the sensor was 4.9 compared with 1.8 detected with the glucometer (p < 0.05). Episodes of mainly nocturnal asymptomatic hypoglycemia were detected with a mean duration of 145 minutes during the night and 75 minutes during the day. CONCLUSIONS: The use of continuous subcutaneous glucose monitoring demonstrates that glycemic objectives are not achieved by conventional insulin therapy. It also shows that there are a high number of hypoglycemic episodes, most of which are asymptomatic. PMID- 12628092 TI - [Values of thyrotropin, free triiodothyronine and free thyroxine using chemiluminescence in children and adolescents in the Autonomous Community of Madrid (Spain)]. AB - OBJECTIVE: To establish the reference ranges of thyrotropin (TSH), free triiodothyronine (FT3) and free thyroxine (FT4) with the chemiluminescence method in healthy children and adolescents in Madrid, Spain. METHOD: A total of 371 children and adolescents of both sexes, with different ages and at different pubertal stage participated in the study. Serum values of TSH, FT3, and FT4 were determined by chemiluminescence. RESULTS: TSH, FT4 and FT3 reference ranges decreased inversely with chronological age. Significant differences were found between prepubertal and pubertal groups. In some groups, there were also occasional differences between sexes. CONCLUSIONS: Thyroid hormone levels decrease with chronological age. Because of the differences in hormone levels between prepubertal and pubertal subjects, we believe that different pubertal stages should be included in the standard reference values of these hormones. PMID- 12628094 TI - [Presence of genu valgum in obese children: cause or effect?]. AB - BACKGROUND: Changes in body configuration that may affect the physical activity may play a role in the caloric consumption and led to the development of obesity. OBJECTIVES: To determine the presence of genu valgum, an alteration that may decrease physical activity and caloric expenditure, in overweight children. METHODS: Thirty-five overweight children without any endocrinological alterations that could lead to obesity were studied. Twenty-nine non-overweight children of a similar age were studied as a control group. In all children weight, height, and body mass index (BMI) were studied, and intermalleolar distance was used to measure the degree of genu valgum. The differences between groups were studied using ANOVA and the correlation between variables was determined using Pearson's correlation. RESULTS: BMI was higher in overweight children than in the control group. Intermalleolar distance was greater in overweight children than in the non overweight group (11.0 0.6 vs 2.90 0.43; p < 0.001). A positive correlation between the intermalleolar distance and the BMI was observed in the overweight group (p < 0.009). Fifty percent of the overweight children showed an intermalleolar distance of more than 10 cm, a value considered abnormal. CONCLUSIONS: The incidence of genu valgum is much higher in overweight children than in non-overweight children of the same age. This alteration may lead to decreased physical activity and lead to obesity. PMID- 12628095 TI - [Immigration: indicators or growth and nutrition]. AB - In the last few years, many immigrant and adopted children from poor countries have come to live in Spain and the number is increasing. In this article we indicate which standard references can help pediatricians to determine the nutritional status and growth of these children whose social, personal and familial histories are different from our own and sometimes unknown. In these cases application of the standard national growth charts of the receiving country is recommended. PMID- 12628093 TI - [Once-daily gentamicin dosing versus thrice-daily dosing in infants with acute pyelonephritis]. AB - BACKGROUND: Once-daily dosing (ODD) of gentamicin is advocated as an effective and safe treatment of Gram-negative bacterial infections in adults. There are insufficient data in the literature to justify its use in infants. OBJECTIVES: To compare the efficacy of ODD of gentamicin with that of classical thrice-daily (t.i.d.) administration in infants with acute pyelonephritis. METHODS: We performed a quasi-experimental study comparing 33 infants who received ODD of gentamicin with a historical control group of 25 infants treated with gentamicin t.i.d. Leukocytosis, C-reactive protein, creatinine, gentamicin dose, peak and trough values, time required for disappearance of fever, and outcome were analyzed. RESULTS: The mean doses of gentamicin (mg/kg/day) were higher in the t.i.d. group (6.4 1.14) than in the ODD group (5.06 0.22; p < 0.001). Peak serum gentamicin concentrations (micro g/ml) were significantly higher in the ODD group (9.32 1.4) than in the t.i.d. group (5.09 1.15; p < 0.001). Mean trough gentamicin concentrations (micro g/ml) were lower in the ODD group than in the t.i.d. group (0.23 0.26 vs 0.78 0.45; p 0.001). There were no significant differences in the duration of fever between the groups (30.64 32 hours in the t.i.d. group vs. 28.57 32 hours in the ODD group). Serum creatinine levels were normal during treatment in both groups. In all patients outcome was good and no adverse effects were noted. CONCLUSIONS: Treatment with ODD of gentamicin in our population of infants with acute pyelonephritis was as effective as traditional administration t.i.d. and possibly was equally safe or safer. PMID- 12628096 TI - [Longitudinal study of the growth of preterm newborn infants]. AB - BACKGROUND: The growth and nutrition of premature infants should be accurately assessed. To do this, reference values of normality, obtained from population to be studied, are required. OBJECTIVES: To study the postnatal growth of premature infants in our environment and to compare their growth with intrauterine growth (Lubchenco et al). PATIENTS: A total of 103 premature infants born at 28-36 weeks of gestation in the Virgen de la Arrixaca University Children's Hospital in Murcia (Spain) from April 1994 to June 1995 were studied. Children who received medical care during the first 24 hours of life and who had no prenatal condition that could affect their growth were selected. Those who suffered from serious disease were excluded. The study was carried out from birth to a postconceptional age of 48-52 weeks. The 2-step regression model was used to calculate the percentile graphs. RESULTS: Percentile growth graphs were drawn up for the following variables: weight, length, arm circumference, head circumference, subscapular fold, and tricipital fold. In premature infants, postnatal growth in weight and length was uniform, progressive, and practically linear and did not show the flattening that is characteristic of intrauterine growth. Other corporal segments such as the head and arm circumference showed a certain slowing down at a postconceptional age of 42 or 43 weeks, and the folds showed a moderate dispersion of values as the study progressed. CONCLUSIONS: Intrauterine growth charts do not accurately assess postnatal growth in premature infants. PMID- 12628097 TI - [Courses in neonatal cardiopulmonary resuscitation]. AB - The optimal management of newborns with asphyxia is closely associated with improved survival and a better quality of life without neuromotor handicaps. Therefore, the training of health professionals who are present at the time of birth in neonatal resuscitation should be a priority. In the present article, we present a program of training courses in neonatal resuscitation. This program has been designed for the training of health care providers and instructors in technical aspects of neonatal resuscitation. The type of courses, their contents and methodology are described. PMID- 12628098 TI - [Childhood immunization schedule of the Spanish Association of Pediatrics 2003]. AB - In this document the Advisory Committee on Vaccines of the Spanish Association of Pediatrics provides recommendations for the immunization schedule for the 2003 2004 season. The use of inactivated poliovirus vaccine is again stressed for children of any age. Moreover, the introduction of the varicella vaccine and the pneumococcal conjugated vaccine, as well as the option of dTpa in adolescents, is highly recommended due to the availability of safe and effective products. Because of the increasing number of new vaccines in the immunization schedule, strategies with combined vaccines must be used. PMID- 12628099 TI - [Knowledge of breastfeeding management among residents in pediatrics]. AB - The staff of maternity wards and clinics for maternal and child health should receive appropriate basic and in-service training on the health benefits of breastfeeding and on lactation management. Pediatricians should not only be knowledgeable about the health, nutritional and physiological aspects of appropriate feeding, they should also be familiar with the mechanics of breastfeeding, its various psychosocial influences, possible difficulties and how to overcome them. To evaluate knowledge of breastfeeding among pediatrics residents throughout Spain, a survey was conducted. A total of 250 questionnaires were collected. Significant differences were observed among provinces. In most areas, residents' training was insufficient. To achieve an appropriate level of knowledge among pediatrics residents in a subject as important to mother and child health as breastfeeding, courses should be given and repeated at regular intervals. Professional associations should be actively involved in promoting appropriate training for health professionals. PMID- 12628100 TI - [Body image and restrictive eating behavior in adolescents]. AB - BACKGROUND: The progressive increase in eating disorders suggests the need to begin health education in early childhood. However, the prevention of eating disorders has been a complicated and, perhaps for this reason, elusive goal for researchers. Restrictive behavior and body image distortion are two basic criteria in the diagnosis of eating disorders. OBJECTIVE: The aim of this study was to explore the relationship between appearance schema and restrictive behavior such as preoccupation with weight and dieting in a random sample of 373 adolescents. METHODS: The instruments used were the Appearance Schemas Inventory (ASI) and the Restraint Scale (RS). Statistical analysis included Pearson's correlation coefficient, Student's t-test, and analysis of variance with the post hoc Tukey contrast test. RESULTS: Body image and restrictive behavior scores were positively correlated (p < 0.0001). Body image distortions were significantly greater in younger adolescents and in those with a higher RS score (i.e. those involved in following restrictive diets and who experienced frequent weight fluctuations) than in older adolescents and in those with a lower RS score. CONCLUSIONS: The results of this study suggest the importance of working in primary prevention and of concentrating efforts on the identification and subsequent modification of body image distortions in children and adolescents. PMID- 12628101 TI - [Cerebrovascular stroke as a sign of atrial myxoma in childhood]. AB - Primary cardiac tumors are uncommon with an estimated incidence of between 0.0017 % and 0.19 %. Most are benign. Whereas myxomas are the most common primary tumor in adults, in children they are exceptionally rare. Cardiac myxomas usually develop in the left atrium, 20 % occur in the right atrium and the remainder develops in the ventricles and rarely in the heart valves. Cerebrovascular strokes secondary to myxoma are rare in childhood. The diagnostic test of choice is transesophageal echocardiogram and early excision is the most effective treatment in preventing serious complications. We report a case of cerebral stroke as the only manifestation of an atrial myxoma in an 11-year-old-girl. The patient presented vertigo, right hemiparesis of the body and dysarthria without loss of consciousness. After diagnostic tests (computerized tomography, magnetic resonance imaging and cerebral angioresonance) she was diagnosed with an ischemic lesion in the left middle cerebral artery. Various investigations were performed to find the cause of the stroke, among them cardiologic study, and a mass in the left atrium suggestive of myxoma was found. The tumor was removed and the diagnosis of myxoma was confirmed by histopathological examination. Outcome was satisfactory. The presence of a cerebral ischemic episode, with or without concomitant heart disease, suggests the need to look for cardiac etiology. PMID- 12628102 TI - [Acromelanosis]. AB - Acromelanosis is an independent disease entity, characterized by increased skin pigmentation, usually located on the acral areas of the fingers and toes. It is mostly seen in newborns or during the first years of life. Only a few cases of this entity have previously been described in the medical literature. In some of these cases, the cutaneous lesions spread to affect large parts of the skin surface. A possible association with other benign and malignant diseases has been proposed. Differential diagnosis must be made with a wide variety of systemic and dermatologic conditions, especially dermatoses with acral distribution of macular hyperpigmentation, including acropigmentation. In this article, we report a new case of acromelanosis in a 5-week-old girl showing two peculiar clinical features: associated melanosis of the genital mucosa and stabilization of the lesions after an initial phase of progression and proximal spread. In addition, the most important features of this rare cutaneous disease are discussed. PMID- 12628103 TI - [Recent progress in donor human milk utilization]. PMID- 12628104 TI - [Acute disseminated encephalomyelitis secondary to Epstein-Barr virus]. PMID- 12628105 TI - [Botulinum toxin in the treatment of psoas muscle contracture in mucopolysaccharidosis]. PMID- 12628106 TI - [Annular alopecia]. PMID- 12628107 TI - [Varicella and stroke]. PMID- 12628109 TI - [Tibial malformation in an infant]. PMID- 12628110 TI - [Impact factor, a strategic goal]. PMID- 12628111 TI - [Survival and clinical features of retinoblastoma]. AB - INTRODUCTION: Retinoblastoma (RB) is the most frequent intraocular tumor in the pediatric age group, representing 3 % of all childhood neoplasms. In Mexico, the annual incidence varies according to the geographical area studied, ranging from 4-24 cases/million in patients younger than 15 years. Ninety-five percent of cases occur in infants and pre-school children. PATIENTS AND METHOD: An observational, retrospective, descriptive, cross-sectional study of all children with RB admitted to the Oncology Department of the Pediatric Hospital of the National Medical Center in a 10-year period was performed. RESULTS: Fifty-six patients were included. The median age was 24 months. Forty-two children presented unilateral RB. Symptoms at diagnosis were leukokoria, strabismus and reduced visual sharpness. Distribution according to Prats' classification was the following: stage I, 5 patients; stage II, 32; stage III, 14; stage IV, 5. Histological analysis revealed 24 patients with poorly differentiated RB, 16 with well-differentiated RB, 13 with moderately differentiated RB and 7 with undifferentiated RB. In eight patients histological type was not identified. In 12 of the 56 patients cytogenetic and molecular studies were performed. All patients underwent surgery. Patients in the early stages were treated with chemotherapy. A total of 26.7 % of the group presented relapse. The disease-free survival at 10 years was 87.5 %. CONCLUSIONS: As with other childhood neoplasms, early diagnosis and multidisciplinary management of RB has improved prognosis and survival. The symptoms, treatment and survival reported in this pediatric cohort are similar to those of other published series. PMID- 12628112 TI - [Survey of the judicious use of antibiotics in primary care]. AB - OBJECTIVE: To identify pediatricians' antibiotic prescribing habits in acute otitis media and tonsillopharyngitis and to determine the interaction between parents and pediatricians concerning antibiotic use in the Autonomous Community of Valencia (Spain). METHOD: Four hundred members of the Valencian Society of Pediatrics were randomly selected. A semi-structured questionnaire with non excluding answers was sent by mail and, when responses were not obtained, a second one was sent. The confidentiality of the information was guaranteed. RESULTS: Of 400 questionnaires sent, 143 (35.8 %) were completed; 88.1 % were completed by pediatricians and 51.1 % by primary care workers. A total of 48.3 % of pediatricians used antibiotics in all cases of acute otitis media and 94.5 % prescribed them when fever and otalgia persisted for more than 48 hours. Amoxicillin-clavulanate was the most frequently prescribed antibiotic (63.6 %). Less than 10 % of pediatricians prescribed antibiotics as empirical treatment in tonsillopharyngitis; amoxicillin was the most frequently prescribed antibiotic (54.6 %). Indications for antibiotic treatment were fever, odynophagia and adenomegaly (69.5 %) and tonsillar exudate (62.5 %). Inappropriate antibiotic use was mainly due to excess workload. Providing health education to parents could be the best way of reducing inappropriate use. CONCLUSIONS: Antibiotic use is frequent in the treatment of acute otitis media. Amoxicillin-clavulanate and amoxicillin were the most frequently prescribed antibiotics in tonsillopharyngitis. Providing health education to parents and reducing pediatricians' workload would decrease inappropriate antibiotic use. PMID- 12628113 TI - [Diagnostic test in emergency departments for bacterial infections in infants younger than 12 months]. AB - OBJECTIVE: To evaluate clinical and analytic numeric data that may help the emergency departments to identify bacterial infections in infants. PATIENTS AND METHODS: A retrospective study of 430 infants with bacterial growth in cultures (culture from blood, 30; urine, 207; stools, 193, and/or cerebrospinal fluid, n 25) was performed. These patients were compared with a control group (n 430), randomly selected from patients aged less than 12 months with negative cultures who were hospitalized with suspected infection. Neonates and surgical patients were excluded from both groups. Statistical analysis was performed using Student's t-test for independent samples, Levene's test for the study of equality of variances, bivariate correlation and one-factor ANOVA, and receiver-operating characteristic (ROC) curves and odds ratios were calculated when statistically significant (p < 0.05) results were obtained. These analyses were performed using the SPSS 10.0 statistical software package. RESULTS: Of the infants admitted to the pediatric unit, 11.7 % had at least one positive bacterial culture. Temperature (p 0.005), leucocyte count (p 0.003), percentage of neutrophils (p < 0.0001) and C-reactive protein (p < 0.0001) were significantly higher in infants with positive cultures. In invasive infections significant differences were found in sex (more frequent in males) (p 0.03), heart rate (p < 0.0001) and respiratory rate (p 0.003). In the ROC curves, the best diagnostic yield was obtained for C reactive protein (0.93 for a cutoff value of 29 mg/l, 86 % specificity and 91 % sensitivity). CONCLUSION: C-reactive protein is essential for diagnosis of bacterial infection in infants in the emergency department. PMID- 12628114 TI - [Emergency department utilization by children aged less than two years old]. AB - BACKGROUND: Emergency department visits are rising although birth rate is decreasing. OBJECTIVE: To compare emergency department.(ED) utilization in the cohorts of children born in 1991 and 1996 in a health district. METHOD: We performed a two-year retrospective cohort study in the health district of Mieres (Spain). All children born in the health district in 1991 (n 600) and 1996 (n 423) were included. The number of visits to the ED was obtained and the frequency and mean number of visits in the first 2 years of life were calculated according to age, sex, and area of residence. RESULTS: The gross number of visits did not vary (1991: 852; 1996: 853), despite a decrease in birth rate. The number of children who attended the ED at least once increased by 34.8 % (1991, 60.6 %; 1996, 82 %) and the frequency of visits increased by 41.7 % (1991, 142.2 %; 1996, 201.4 %). More than 60 % of visits were made by 20 % (1991) of the children and by 29 % (1996). The percentage of children who attended the ED, the frequency rate and the mean number of visits were significantly higher in the municipality nearest the hospital than in the remaining municipalities in the health district. CONCLUSIONS: Although the gross number of visits to the ED remained unchanged, ED utilization increased substantially in only 5 years. PMID- 12628115 TI - [Hyperfrequent consultations: is there a relationship with the mother's personality?]. AB - OBJECTIVES: To study the psychological profile of mothers who overuse pediatric services and associated social and demographic characteristics, as well as to evaluate the association between maternal personality profile and greater use of pediatric services. MATERIAL AND METHOD: Cross-sectional descriptive study, at the Basic Health Areas of Granollers Sud and Granollers Oest-Canovelles in Barcelona (Spain). SUBJECTS: Forty-four mothers, representing 21.5 % of the population meeting the inclusion criteria were studied. The population consisted of mothers of children who visited the center on an equal or higher number of occasions as two standard deviations. PROCEDURE: One-hour interviews were conducted. A data sheet to assess social and demographic variables, Milton's Multiaxial Inventory II, and Goldberg's Anxiety and Depression Scale to detect symptoms of depression and anxiety were administered. RESULTS: Forty-four mothers (21.5 %) attended the interviews. The mean age was 36.6 years. Most of the mothers (90.9 %) were married, 73 % had completed primary studies, 5 % were housewives, and 75 % had no family support. Fifty percent of the children were the youngest in their family, and 30 % were only children. Concerning personality disorders, 55.8 % of overusing mothers showed compulsive personality, 30.23 % schizoid personality, and 14 % dependent personality while 47.7 % of mothers presented significant symptoms of anxiety. CONCLUSIONS: Sociodemographic profile, anxiety, and maternal personality profile are related to overuse of pediatric services. Compulsive, schizoid and dependent personality disorders seem to be associated with overuse. PMID- 12628116 TI - [Somatization symptoms. An emerging pediatric entity]. AB - BACKGROUND: Somatization symptoms are a clinical reality in our environment. However, many pediatricians have little information about this condition or experience of its management. OBJECTIVE: To determine the clinical and differential characteristics of these patients. The early identification of these patients and initiation of therapy in the initial stages of the process would improve prognosis. MATERIAL AND METHOD: A retrospective review was performed of the children admitted to the short-stay unit of a tertiary hospital because of somatic complaints and whose final diagnosis was that of a somatization disorder. RESULTS: Sixty medical records were analyzed, of which 38 (63 %) corresponded to girls, with a mean age of 11 years at presentation. The most frequent reasons for consultation were related to the digestive and neurological systems. Thirty-four patients (57 %) had previously consulted for the same reason. In the sample analyzed, the most frequent personality trait was anxiety. The main triggers were familial and school factors. The most frequent diagnosis was pain disorder in 42 children (70 %). All patients received psychotherapy and 39 received complementary pharmacological treatment. CONCLUSIONS: The data analyzed in this study indicate that somatization symptoms most frequently occur in anxious, prepubescent girls, with migraine or non-specific abdominal pain of approximately one month's duration. Patients have usually made several previous visits and no organic causes are discovered on physical examination. PMID- 12628117 TI - [Complementary therapies in cystic fibrosis: evidence of therapeutic benefits and treatment recommendations]. AB - Cystic fibrosis (CF) is an autosomal-recessive disorder that predominantly affects the respiratory system. When this disease was described in 1938 the mortality rate was approximately 70 % in the first year of life. Survival has dramatically increased from a median of approximately 4 years in the 1960s to 19 years in the 1970s and 33 years in 2001 according to figures from the American Cystic Fibrosis Foundation. This impressive increase in the life expectancy of individuals with CF is undoubtedly related to recent advances in the organization of specialized CF units and to the use of new therapies against respiratory involvement.The traditional basis of treatment for CF lung disease includes nutritional support, antibiotic therapy, chest physical therapy and aerobic exercise. Preventive measures such as influenza vaccination and avoidance of tobacco smoke are also useful. Several new approaches such as ion transport therapy, protein therapy and gene therapy are currently being developed. Many studies have provided clear evidence of the therapeutic benefits of antibiotics, respiratory physiotherapy, exercise, and nutrition. In this article we review the scientific evidence on the advantages of the use of several therapeutic interventions against inflammation, increased sputum viscoelasticity and adhesiveness, and bronchial obstruction in CF patients. PMID- 12628118 TI - [The Europe against immature lung (EURAIL) project. Strategies for the prevention and treatment of lung immaturity in neonates]. AB - OBJECTIVE: To determine clinical practice variability in the prevention and postnatal treatment of lung immaturity in Spain. PATIENTS AND METHODS: Cross sectional study within a larger study in 13 European countries. Data were obtained from the medical records of all very low birth weight (VLBW) infants born in participating centers, without other instrumentation. RESULTS: A questionnaire was sent to 213 centers. Seventy-two (34 %) responded, with 162,157 births (40 % of total births in 1999). Eight percent of infants were of low birth weight, 1.2 % (2,015) of very low birth weight and 0.45 % were of extremely low birth weight. A total of 52.2 % of VLBW infants received at least one dose of prenatal steroids, 28.8 % received a full course and 9.3 % received more than one cycle. All centres used surfactant and 76.8 % had a written protocol. Forty-one percent of VLBW infants were intubated, 47.6 % required mechanical ventilation for more than 6 hours, and 5 % underwent continuous positive airway pressure. A total of 80.4 % used postnatal steroids, half of them for chronic lung disease prevention, and 83.4 % used steroids to treat this disease. Steroids were most frequently indicated at 7-14 days of life for 3-9 days. The most important causes of neonatal morbidity were chronic lung disease in 14 %, ductus arteriosus in 16.7 %, intraventricular hemorrhage in 8.5 %, and necrotizing enterocolitis in 7.3 %. CONCLUSIONS: Prenatal exposure to steroids was low. Repeat cycles and postnatal steroid use to prevent chronic lung disease was high. Recent scientific evidence on the use of pre- and postnatal steroids should be more widely disseminated. PMID- 12628120 TI - [Pneumonectomy in cystic fibrosis]. AB - We present the case of a 13-year-old boy with cystic fibrosis (CF) who developed severe right-sided lung infection with formation of abscess and localized bronchiectasis. The boy's lung disease was complicated by nephrotic syndrome and secondary amyloidosis. Unilateral pneumonectomy was performed, producing significant clinical improvement with a remarkable increase in quality of life which has lasted to the present date, 15 years later. Most patients with CF develop lung disease, which is the main cause of adult mortality in this population. Lung transplantation is currently considered the treatment of choice in severe bilateral lung disease in CF. However, in severe unilateral lung disease such as localized bronchiectasis, surgical resection of the affected lobe or lung is still a worthwhile option as a rescue therapy for patients who are at high risk of dying while waiting for lung transplantation. PMID- 12628119 TI - [Congenital adrenal hyperplasia: an association with increased fetal nuchal translucency]. AB - A neonate with increased nuchal translucency and congenital adrenal hyperplasia is described. The possible interferences in hormone assays when values are much higher than the average assay range are also discussed. PMID- 12628121 TI - [Acute liver failure due to T cell lymphoma without hepatic infiltration]. AB - Hepatomegaly and alterations in hepatic function are common to all patients with sickle-cell disease. In these patients, hepatic sickling is a manifestation of severe intrahepatic vaso-oclusive crises, even at levels of 25 % HbS and hematocrits of more than 45-50 %, which in 10 % of cases can lead to acute hepatic failure (AHF). AHF can be due to a variety of causes, including hematologic malignancies, but T cell lymphoma, which is usually secondary to diffuse hepatic infiltration and ischemia, is an exceptional cause, although other mechanisms can be involved. Cytokines released by lymphomas have recently been implicated as a cause of AHF.We describe a black girl with sickle cell disease, who developed AHF due to T cell lymphoma without lymphomatous infiltration of the liver. The only mechanism found to explain the clinical findings was release of cytokines by lymphoma. In patients with AHF of unknown etiology we propose early liver biopsy, because prognosis depends on the presence or absence of hepatic tumour infiltration. If AHF develops in a patient with diagnosed malignant disease, cytokine release may be the cause of AHF. Consequently, early diagnosis of the underlying disease and provision of liver support, as well as direct removal of inflammatory mediators from the circulation by exchange transfusion or other methods, should be the main priorities. PMID- 12628122 TI - [Graves' disease in patients with Down syndrome]. AB - Three patients showing the rare association of Down syndrome and Graves' disease are reported. While two of the patients were asymptomatic, the third showed goiter, nervousness, weight loss, and tachycardia. In addition to the typical features of hyperthyroidism, this patient showed right heart failure and hypertransaminasemia, which disappeared with antithyroid treatment. Because Graves' disease is rare in children, and the clinical presentation was unusual in one of our patients, we report three patients with Graves' disease and Down syndrome, and emphasize the importance of periodic evaluation of thyroid function in children with Down syndrome not only to detect hypothyroidism. PMID- 12628123 TI - [Gynecomastia secondary to Leydig cell tumor]. AB - We describe a 16-year-old boy with Leydig cell tumor who initially presented bilateral gynecomastia with increased estradiol concentrations, decreased testosterone concentrations and normal gonadotropin levels. Testicular ultrasonography showed a tumor in the left testicle, and orchidectomy was performed. Histopathological analysis revealed a Leydig cell tumor. Two months after surgery, the gynecomastia diminished gradually and estrogen levels returned to normal. No recurrences have occurred during a 2-year follow-up. PMID- 12628124 TI - [Coronary ischemia secondary to congenital anomaly of the left coronary artery]. AB - Coronary artery anomalies constitute 2.2 % of congenital malformations of the heart. The most common abnormality is anomalous origin of the left coronary artery from the pulmonary trunk, also known as Bland-White-Garland syndrome. Clinical manifestations are due to myocardial ischemia caused by the creation of an arteriovenous shunt. The childhood type of this anomaly presents high mortality from heart failure. The adult type develops myocardial infarction, arrhythmias, sudden cardiac death or signs of congestive heart failure. Surgical repair is essential. Various surgical approaches are available and the treatment of choice is direct left coronary artery reimplantation in the aorta. An alternative technique is to create an aortopulmonary tunnel (Takeuchi technique). Marked improvement is usually observed after surgical repair. We report two cases of myocardial ischemia due to coronary anomalies. PMID- 12628125 TI - [Prolonged Guillain-Barre syndrome]. AB - Outcome in Guillain-Barre syndrome (GBS) in childhood is usually favorable although prolonged and severe forms may develop. We report the case of a 12-year old boy with rapidly ascending, slowly-progressive GBS with severe cranial nerve and autonomic dysfunction. Diagnosis was based on clinical presentation, electromyography, cerebrospinal fluid analysis and plasmatic antiganglioside GM1 antibodies. Early treatment with an initial loading dose of 2 g/kg of intravenous gammaglobulins over 5 days was given with a significant but slow clinical improvement. A repeat infusion of gammaglobulins was given two months later. Response was satisfactory with motor, sensory and autonomic dysfunction recovery. A third course of immunoglobulins 28 days later produced little benefit. At the present time, 5 months after the onset of the disease, flaccid paralysis of the lower limbs and the distal portion of the upper limbs persists but autonomic instability has disappeared. The patient has been decannulated and is following an ambulatory rehabilitation program. Although repeated intravenous immunoglobulins may be useful in the treatment of prolonged GBS, the key issue is excellent intensive care unit management. Further research examining potentially more effective treatment such as beta-interferon and immunosuppressive agents is required. PMID- 12628126 TI - [Bilateral facial paralysis as the first manifestation of Guillain-Barre syndrome]. PMID- 12628128 TI - [Variceal ligation in pediatric patients]. PMID- 12628127 TI - [Congenital pseudoarthrosis of the clavicle in twins]. PMID- 12628130 TI - [Sudden loss of consciousness and comitial crisis after head injury in a teenager]. PMID- 12628129 TI - [Nitroprusside poisoning]. PMID- 12628131 TI - [Hypertrophic and redundant Eustachian valve]. PMID- 12628132 TI - [Is cardiological evaluation enough in Fallot's tetralogy?]. PMID- 12628134 TI - [Asthma and respiratory syncytial virus]. PMID- 12628137 TI - [Sickle-cell anemia: an emerging disease in Spain]. PMID- 12628136 TI - [Bone age delay and hypoplastic thumbs in a five year-old boy]. PMID- 12628138 TI - [Sickle cell anemia: experience in a center]. AB - BACKGROUND: Sickle cell anemia is a structural hemoglobinopathy in which morphological and physical changes in erythrocytes cause vaso-occlusive episodes in various organs and tissues. The disease is common among blacks and the African population. As a result of the growing migratory flow, this is an emerging disease in Spain. OBJECTIVE: To present the casuistics of a pediatric hospital: clinical onset, the most frequent features and complications, and treatment. MATERIAL AND METHODS: We performed a retrospective study of 22 patients aged less than 18 years old diagnosed with sickle cell anemia between January 1985 and December 2001. Epidemiologic data, symptoms, complications, blood test results, treatment, and response were recorded. RESULTS: The mean age of the patients was 39 months. In 54 %, diagnosis was established before the age of 2 years. No differences were found in sex. The countries of origin were Gambia in 32 %, Morocco in 23 %, and Senegal in 18 % as well as other African and Central America countries; 53 % of the children were born in Spain. The most common complaint was vaso-occlusive pain localized in the abdomen (45 %). The most frequent complications were infections and 13.7 % suffered stroke. Twenty-eight percent of the patients diagnosed before the age of 2 years presented complications. Eleven patients received hydroxyurea for recurrent vaso-occlusive crises with favorable results; one patient underwent splenectomy and another received an allogenic bone marrow transplant from an HLA-identical brother with excellent results. CONCLUSIONS: This study reproduces the data described in the literature from countries with a high prevalence of the disease. Morbidity could be minimized by early diagnosis and preventive treatment and good healthcare. Given the increasing incidence of the disease, screening of black and African neonates and genetic counseling are recommended together with guidelines for prompt and appropriate treatment in primary health centers and emergency departments. PMID- 12628139 TI - [Incidence of allergy to cow's milk protein in the first year of life and its effect on consumption of hydrolyzed formulae]. AB - OBJECTIVES: The aims of this study were to determine the incidence of allergy to cow's milk protein in infants and the cost of treatment with hydrolyzed formulae in the Autonomous Community of Madrid. PATIENTS AND METHODS: Infants with suspected adverse reaction to cow's milk protein born between March 1, 2000 and February 28, 2001 within the catchment area covered by the pediatric services of Hospital La Paz were studied. The diagnostic algorithm of allergy to cow's milk protein of our department was used. The theoretical consumption of hydrolyzed formulae in the treatment of allergic infants was calculated and extrapolated to the Community of Madrid. RESULTS: There were 5367 live births in the above mentioned catchment area. Adverse reaction to cow's milk protein was suspected in 185 infants. Allergy to cow's milk protein was confirmed in 101 (54 %). The incidence of allergy to cow's milk protein was 101/5356 (1.9 %). In the same period there were 57 078 live births in the Autonomous Community of Madrid. The number of infants that might be allergic in one year would be 1084. The cost of hydrolyzed formulae per infant and year would be 1,585.72 Euros. Thus, the cost of hydrolyzed formulae in the 1084 allergic infants would be 1,718,922.9 Euros; in the 923 in whom allergy was ruled out, the cost would be 1,463,621.8 Euros. CONCLUSIONS: The incidence of allergy to cow's milk protein in the first year of life is at least 1.9 %. The correct diagnosis allows us to rule out allergy to cow's milk protein in almost half of the cases, thus avoiding the use of unnecessary substitutive diets involving a high cost. PMID- 12628140 TI - [Effects of prone position, inhaled nitric oxide and surfactant in children with hypoxemic pulmonary disease]. AB - OBJECTIVE: To analyze the therapeutic response to prone position, inhaled nitric oxide (NO) and surfactant in children with hypoxemic pulmonary disease. PATIENTS AND METHODS: We studied the effect of prone position, NO, and surfactant in critically ill children with acute hypoxemic pulmonary disease unresponsive to conventional therapy. We analyzed PaO2, SatO2, the PaO2/FiO2 ratio, oxygenation index and PaCO2 before and after each treatment, as well as the subsequent clinical course. An increase of more than 20 % in the PaO2/FiO2 ratio was considered a positive response. RESULTS: Ninety treatments were administered in 56 patients: 55 patients were treated with NO, 18 with prone position and 17 with surfactant. All three treatments substantially improved oxygenation. The mean increase in the PaO2/FiO2 ratio was 35 % with nitric oxide, 33 % with prone position and 50 % with surfactant. The mean decrease in oxygenation index was 22 % with nitric oxide, 24 % with prone position and 17 % with surfactant. Seventy one percent of patients treated with NO, 61 % of patients treated with prone position, and 64 % of patients who received surfactant were responders. The three treatments produced a slight decrease in PaCO2 (2.5 mmHg with nitric oxide, 4.7 mmHg with prone position and 5.1 mmHg with surfactant). CONCLUSIONS: Inhaled NO, prone position and surfactant improve oxygenation in some children with hypoxic pulmonary disease. PMID- 12628141 TI - [Pre and postnatal tobacco exposure and bronchiolitis]. AB - OBJECTIVES: To determine the influence of pre- and postnatal tobacco exposure in the development of bronchiolitis. PATIENTS AND METHODS: A questionnaire was given to the parents of children hospitalized between August 2001 and August 2002. It included items on parental smoking habits and maternal smoking during pregnancy. Clinical, analytical and radiological criteria were used to diagnose bronchiolitis. RESULTS: Of 450 children, 123 (27.3 %) were diagnosed with bronchiolitis. The control group was composed of 327 children. A total of 61.6 % of the children had at least one parent who smoked and 32.9 % of these children developed bronchiolitis; 39.1 % had non-smoking parents and 18.2 % were hospitalized with bronchiolitis (OR 2.20 [1.39-3.47]). Of the entire group of children studied, 35.3 % had mothers who smoked and 37.7 % of these children had bronchiolitis compared with 21.4 % of children whose mothers were non-smokers (OR 2.22 [1.45-3.39]). A total of 49.6 % had fathers who smoked, and 32.3 % of these children were diagnosed with bronchiolitis compared with 22 % of children whose fathers were non-smokers (OR 1.65 [1.10-2.57]). Forty-four percent of children whose mothers smoked during pregnancy were hospitalized with bronchiolitis compared with only 20.9 % of non-exposed children (OR 2.96 [1.90-4.62]). The large number of mothers who smoked during pregnancy (27.8 %) was notable. Multivariant analysis with logistic regression was performed and the only variable that remained statistically significant was smoking during pregnancy (p < 0.00001; OR 3.27 [1.39-7.71]). CONCLUSIONS: Maternal smoking during pregnancy seems to be the main risk factor for the subsequent development of bronchiolitis PMID- 12628142 TI - [Childhood cancer in the Autonomous Community of Cantabria in Spain (1995-2000)]. AB - BACKGROUND: Since 1980, the epidemiology of childhood cancer in Spain has been registered through the National Registry of Childhood Cancer. However, this registry does not include patients from Autonomous Community of Cantabria because there is no reporting clinical center. The absence of data on childhood cancer in this region justifies this study. OBJECTIVES: To analyze the clinical presentation, diagnostic delay and incidence of childhood cancer in Cantabria. METHODS: We performed a retrospective analysis of 89 children (aged 0-15 years) diagnosed with cancer in Cantabria from 1995-2000. RESULTS: The annual incidence of childhood cancer in the region was 198.1 cases per million inhabitants with a predominance of males (53.9 %) and children aged less than 5 years (46.1 %). The most frequent cancers were leukemia (32.6 %) and brain tumors (23.6 %). The most frequent signs and symptoms were fever (29.2 %) and decreased appetite or fatigue (19.1 %). The mean delay in diagnosis for all tumors was 5.99 weeks and delays were longest for brain tumors. There was a direct statistical relationship between non-specific clinical presentation and diagnostic delay. A total of 20.2 % of all patients received treatment outside Cantabria. CONCLUSIONS: The incidence of all childhood cancers and especially that of neuroblastoma was higher in Cantabria than in other areas of Spain. In general, the major difficulties in the diagnosis of childhood cancer are its low incidence and non specific presenting symptoms. Specialized pediatric oncology units near to patients' homes should be created to avoid problems due to treatment outside the area of residence. PMID- 12628143 TI - [Children and adolescents in foster care: health problems and guidelines for their health care]. AB - In Spain, between 8,000 and 10,000 children and adolescents enter foster or residential care every year. This article aims to provide a review to increase knowledge of the health problems of minors in foster care.Sixty-five percent of the studies consulted were performed in the USA, 25 % in Spain and 10 % in other developed and industrialized countries. These studies report that a high percentage of these minors present complicated and serious physical, mental, and/or developmental problems. However, no appreciable qualitative differences in the most frequent health problems presented by these minors have been observed and there is general consensus that the high-priority health needs are the provision of preventive and/or therapeutic psychopedagogic, psychiatric, dermatologic, dental, nutritional, ophthalmologic, respiratory and immunization services. Failure to identify and provide early treatment of the health needs of these minors not only adversely affects their quality of life and future physical, emotional and intellectual development, but can also increase their difficulties in adaptation while in foster care and their future social adaptation. Their permanent relationship with the biological or adoptive family when foster care stops could also be jeopardized. Therefore, all children and adolescents in foster care should receive initial health screenings, comprehensive assessments and monitoring of their physical, mental health, and developmental status. We provide guidelines for the healthcare of these minors, which should be of use to healthcare professionals taking care of these children and adolescents while they remain in foster care. PMID- 12628144 TI - [Antiepileptic drugs used in childhood. New products and new concepts]. AB - INTRODUCTION: Eight new antiepileptic drugs (AED) have been marketed in Spain since 1990 and others will soon follow. OBJECTIVE: To review the concepts underlying the development of the new drugs, as well as their indications, efficacy and safety. DEVELOPMENT: Pharmacologic antiepileptic intervention is no longer solely directed towards an anticonvulsant effect, but also to epileptogenic prevention, disease modification and reversal of pharmacoresistance. The development of new AED, initially based on the screening of putative products in animal models, changed during the last half of the century to a rational design based on known facts about excitatory /inhibitory neuronal mechanisms. More recently, attention has focussed on pharmacogenetics. The new AED were initially indicated for partial epilepsies, but some have been shown to have a broader clinical spectrum. Some show the ideal pharmacokinetic mechanisms, avoiding hepatic metabolism and protein binding. Drug interactions and adverse effects, especially severe idiosyncratic adverse effects, are rare, although there are some exceptions. In most cases, however, seizure control does not seem to be better than with the classic AED. Because of the specific characteristics of childhood epilepsy and pharmacokinetics, as well as the regulations governing the development of clinical trials, the use of new products in children is circumspect, which in turn delays the access of such patients to a possible therapeutic benefit. PMID- 12628145 TI - [Early detection of sickle cell anemia and other hemoglobinopathies in neonates in the Autonomous Community of Madrid. A pilot study]. AB - OBJECTIVE: To determine the incidence of sickle cell anemia and other hemoglobinopathies in the neonatal population of the Autonomous Community of Madrid and to determine the need for a screening program. METHODS: The study was performed with the same blood spot specimen dried on filter paper used for congenital hypothyroidism and congenital adrenal hyperplasia screening. All neonates born in the public and private hospitals of the Autonomous Community of Madrid were included and universal-type screening was performed. High-performance liquid chromatography (HPLC) was used to detect variant hemoglobins. The variant automated system was used to separate and identify hemoglobin F, A1c, A, S, C, A2/E and D. To confirm variant hemoglobins, specific HPLC for -thalassemia (ion exchange) and globin chains (reversed phase) with a more expanded gradient were used. RESULTS: A total of 29 253 specimens were screened and 98 cases of variant hemoglobins were detected. The overall incidence was 1/299. There were five cases of sickle cell disease (HbFS and HbFS(tal), with an incidence of 1/5.851, and 71 cases of sickle cell traits (1/412). CONCLUSIONS: These results confirm the need to include screening for sickle cell disease and other hemoglobinopathies in our neonatal program. PMID- 12628146 TI - [Retinopathy of prematurity: incidence, severity and outcome]. AB - OBJECTIVE: The aim of this study was to evaluate the incidence, severity and outcome of retinopathy of prematurity (ROP) in neonates with birth weights of less than 1,501 g in our environment. METHODS: A prospective study of infants with birth weights of less than 1,501 g was performed from January 1999 to June 2000. One hundred seventy-eight infants were enrolled in the study. Mean gestational age was 29.8 2.9 weeks and mean birth weight was 1,095 261.5 g. RESULTS: Overall, 52 infants (29.2 %) developed ROP, of which 10 (5.6 %) presented threshold ROP requiring treatment. None of the infants with a gestational age of more than 30 weeks and a birth weight of more than 1,250 g presented severe retinopathy. CONCLUSIONS: The incidence found in the present study is similar to that of other studies. Immaturity is the most significant risk factors for the development of ROP. The results of our study led us to change our screening program to detect all threshold ROP. PMID- 12628147 TI - [Cholestasis in neonates and infants. A diagnostic guide]. PMID- 12628148 TI - [Biliary atresia]. PMID- 12628149 TI - [Medical treatment and nutritional support in patients with chronic cholestatic disease]. PMID- 12628150 TI - [Pericardial defect with atrial herniation]. AB - Pericardial defects are rare in childhood and outcome is usually benign. Patients can be asymptomatic, but they may experience vague chest pain, angina, myocardial ischemia, emboli, and arrhythmia and some cases of sudden death have even been described in the literature. We report the case of a 3-month-old girl who, in the context of an episode of fever, underwent chest X-ray examination, which showed a left heart border anomaly. This finding strongly suggested a congenital partial pericardial defect, which was subsequently confirmed by other techniques. The interest of this case lies in the ability of the pediatrician to establish a presumptive diagnosis by examining the chest X-ray. We conclude by stressing the need for a high degree suspicion for congenital heart anomalies because of the life-threatening nature of some partial pericardial defects, which can lead to left-ventricular herniation. PMID- 12628152 TI - [Parathyroid cyst in the differential diagnosis of neck masses. A case report]. AB - Parathyroid cysts are rarely seen in clinical practice. They should always be included in the differential diagnosis of neck masses. Conclusive diagnosis should be established by fine-needle aspiration (FNA), a procedure that is useful for both diagnostic and therapeutic purposes.We report the case of girl who aroused clinical suspicion of thyroid node. However, the diagnosis of parathyroid cyst was subsequently confirmed by FNA. PMID- 12628151 TI - [Persistence of antibodies three years after vaccination with the 23-valent pneumococcal vaccine]. AB - OBJECTIVE: To study the persistence of antibodies after primary immunization with the 23-valent pneumococcal vaccine in children with nephrotic syndrome. METHODS: Sera from 14 of 26 children with nephrotic syndrome were obtained 3 years after vaccination. We used an ELISA method to measure IgG antibody levels to pneumococcal serotypes 14, 23F, 3 and 6B. Antibody levels before vaccination, 1 month and 3 years after vaccination were compared. RESULTS: Significant increases in specific antibody concentrations were observed 30 days after vaccination for all serotypes except serotype 3. Differences in response according to serotype were found. The highest serological response was observed for serotype 14 (78.5 % of the patients showed a good-to-moderate response). Three years after vaccination antibody levels were significantly decreased (27.3 % of children for serotype 14, 46.2 % for serotype 23F and 50 % for serotype 6B). CONCLUSIONS: The 23-valent pneumococcal vaccine is immunogenic in children aged more than 2 years with nephrotic syndrome. Response varied according to serotype. Persistence of vaccine-induced antibodies is short-term, with low antibody levels 3 years after immunization. The results of this study suggest that these patients require revaccination 3 years after the first dose. PMID- 12628153 TI - [Chronic relapsing pancreatitis]. PMID- 12628154 TI - [Intestinal subocclusion due to congenital trichobezoar]. PMID- 12628155 TI - [Cerebral candidiasis in a neonate]. PMID- 12628156 TI - [Global transitory amnesia: an adult disorder also present in childhood]. PMID- 12628157 TI - [Astrovirus in the etiology of pediatric gastroenteritis]. PMID- 12628158 TI - [Conservative treatment of phimosis, balanitis and perianal streptococcal disease with local steroid therapy]. PMID- 12628159 TI - [Spontaneous splenic rupture secondary to infectious mononucleosis]. PMID- 12628160 TI - [New indication for pneumococcal vaccines]. PMID- 12628161 TI - [Episode of hypotonia and weakness of the lower extremities in two siblings]. PMID- 12628162 TI - Molecular identification of the Na+-activated K+ channel. AB - Progress in understanding sodium-activated potassium channels (K(Na)), suggested to function in excitable cells both during physiological conditions and protectively during hypoxia, has been limited by their unknown molecular identity. In this issue of Neuron, Salkoff and coworkers now show that members of the Slo gene family, Slo2.1 and Slo2.2, encode functional K(Na) channels. PMID- 12628163 TI - Bassoon's part in two presynaptic orchestras. AB - The protein Bassoon is found in the cytoskeletal matrix at the active zone of conventional synapses and in presynaptic ribbons of photoreceptor synapses. Two new studies in Neuron show that Bassoon's most prominent role at conventional synapses is to enable vesicle cycling, whereas, in photoreceptors it attaches the ribbon to the presynaptic membrane. PMID- 12628164 TI - Molecular mechanisms of mechanosensation: big lessons from small cells. AB - Little is known of molecular mechanisms of human mechanosensation. Only now are candidate eukaryotic sensors being identified. In contrast, bacterial sensors, including mechanosensitive channels, have been cloned, sequenced, reconstituted, and functional mutants characterized. Moreover, crystal structures for bacterial mechanosensitive channels have been resolved and structural gating transitions predicted. These studies give clues to general principles underlying the ability of a membrane protein to sense and respond to perturbations of its lipid environment that may be conserved between bacteria and humans. PMID- 12628165 TI - Parkin is a component of an SCF-like ubiquitin ligase complex and protects postmitotic neurons from kainate excitotoxicity. AB - Mutations in parkin, which encodes a RING domain protein associated with ubiquitin ligase activity, lead to autosomal recessive Parkinson's disease characterized by midbrain dopamine neuron loss. Here we show that parkin functions in a multiprotein ubiquitin ligase complex that includes the F-box/WD repeat protein hSel-10 and Cullin-1. HSel-10 serves to target the parkin ubiquitin ligase activity to cyclin E, an hSel-10-interacting protein previously implicated in the regulation of neuronal apoptosis. Consistent with the notion that cyclin E is a substrate of the parkin ubiquitin ligase complex, parkin deficiency potentiates the accumulation of cyclin E in cultured postmitotic neurons exposed to the glutamatergic excitotoxin kainate and promotes their apoptosis. Furthermore, parkin overexpression attenuates the accumulation of cyclin E in toxin-treated primary neurons, including midbrain dopamine neurons, and protects them from apoptosis. PMID- 12628166 TI - Neuronal or glial progeny: regional differences in radial glia fate. AB - The precursor function of the ubiquitous glial cell type in the developing central nervous system (CNS), the radial glia, is largely unknown. Using Cre/loxP in vivo fate mapping studies, we found that radial glia generate virtually all cortical projection neurons but not the interneurons originating in the ventral telencephalon. In contrast to the cerebral cortex, few neurons in the basal ganglia originate from radial glia, and in vitro lineage analysis revealed intrinsic differences in the potential of radial glia from the dorsal and ventral telencephalon. This shows that the progeny of radial glia not only differs profoundly between brain regions but also includes the majority of neurons in some parts of the CNS. PMID- 12628167 TI - The sodium-activated potassium channel is encoded by a member of the Slo gene family. AB - Na(+)-activated potassium channels (K(Na)) have been identified in cardiomyocytes and neurons where they may provide protection against ischemia. We now report that K(Na) is encoded by the rSlo2 gene (also called Slack), the mammalian ortholog of slo-2 in C. elegans. rSlo2, heterologously expressed, shares many properties of native K(Na) including activation by intracellular Na(+), high conductance, and prominent subconductance states. In addition to activation by Na(+), we report that rSLO-2 channels are cooperatively activated by intracellular Cl(-), similar to C. elegans SLO-2 channels. Since intracellular Na(+) and Cl(-) both rise in oxygen-deprived cells, coactivation may more effectively trigger the activity of rSLO-2 channels in ischemia. In C. elegans, mutational and physiological analysis revealed that the SLO-2 current is a major component of the delayed rectifier. We demonstrate in C. elegans that slo-2 mutants are hypersensitive to hypoxia, suggesting a conserved role for the slo-2 gene subfamily. PMID- 12628168 TI - The presynaptic active zone protein bassoon is essential for photoreceptor ribbon synapse formation in the retina. AB - The photoreceptor ribbon synapse is a highly specialized glutamatergic synapse designed for the continuous flow of synaptic vesicles to the neurotransmitter release site. The molecular mechanisms underlying ribbon synapse formation are poorly understood. We have investigated the role of the presynaptic cytomatrix protein Bassoon, a major component of the photoreceptor ribbon, in a mouse retina deficient of functional Bassoon protein. Photoreceptor ribbons lacking Bassoon are not anchored to the presynaptic active zones. This results in an impaired photoreceptor synaptic transmission, an abnormal dendritic branching of neurons postsynaptic to photoreceptors, and the formation of ectopic synapses. These findings suggest a critical role of Bassoon in the formation and the function of photoreceptor ribbon synapses of the mammalian retina. PMID- 12628170 TI - Probing the intracellular calcium sensitivity of transmitter release during synaptic facilitation. AB - In nerve terminals, residual Ca(2+) remaining from previous activity can cause facilitation of transmitter release by a mechanism that is still under debate. Here we show that the intracellular Ca(2+) sensitivity of transmitter release at the calyx of Held is largely unchanged during facilitation, which leaves an increased microdomain Ca(2+) signal as a possible mechanism for facilitation. We measured the Ca(2+) dependencies of facilitation, as well as of transmitter release, to estimate the required increment in microdomain Ca(2+). These measurements show that linear summation of residual and microdomain Ca(2+) accounts for only 30% of the observed facilitation. However, a small degree of supralinearity in the summation of intracellular Ca(2+) signals, which might be caused by saturation of cytosolic Ca(2+) buffer(s), is sufficient to explain facilitation at this CNS synapse. PMID- 12628169 TI - Functional inactivation of a fraction of excitatory synapses in mice deficient for the active zone protein bassoon. AB - Mutant mice lacking the central region of the presynaptic active zone protein Bassoon were generated to establish the role of this protein in the assembly and function of active zones as sites of synaptic vesicle docking and fusion. Our data show that the loss of Bassoon causes a reduction in normal synaptic transmission, which can be attributed to the inactivation of a significant fraction of glutamatergic synapses. At these synapses, vesicles are clustered and docked in normal numbers but are unable to fuse. Phenotypically, the loss of Bassoon causes spontaneous epileptic seizures. These data show that Bassoon is not essential for synapse formation but plays an essential role in the regulated neurotransmitter release from a subset of glutamatergic synapses. PMID- 12628171 TI - Lurcher GRID2-induced death and depolarization can be dissociated in cerebellar Purkinje cells. AB - The Lurcher mutation transforms the GRID2 receptor into a constitutively opened channel. In Lurcher heterozygous mice, cerebellar Purkinje cells are permanently depolarized, a characteristic that has been thought to be the primary cause of their death, which occurs from the second postnatal week onward. The more dramatic phenotype of Lurcher homozygotes is thought to be due to a simple gene dosage effect of the mutant allele. We have analyzed the phenotype of Lurcher/hotfoot heteroallelic mutants bearing only one copy of the Lurcher allele and no wild-type Grid2. Our results show that the absence of wild-type GRID2 receptors in these heteroallelic mutants induces an early and massive Purkinje cell death that is correlated with early signs of autophagy. This neuronal death is independent of depolarization and can be explained by the direct activation of autophagy by Lurcher GRID2 receptors through the recently discovered signaling pathway formed by GRID2, n-PIST, and Beclin1. PMID- 12628173 TI - Integrating the molecular and cellular basis of odor coding in the Drosophila antenna. AB - We investigate how the molecular and cellular maps of the Drosophila olfactory system are integrated. A correspondence is established between individual odor receptors, neurons, and odors. We describe the expression of the Or22a and Or22b receptor genes, show localization to dendritic membranes, and find sexual dimorphism. Or22a maps to the ab3A neuron, which responds to ethyl butyrate. Analysis of a deletion mutant lacking Or22a, along with transgenic rescue experiments, confirms the mapping and demonstrates that an Or gene is required for olfactory function in vivo. Ectopic expression of Or47a in a mutant cell identifies the neuron from which it derives and its odor ligands. Ectopic expression in a wild-type cell shows that two receptors can function in a single cell. The ab3A neuron does not depend on normal odor receptor gene expression to navigate to its target in the CNS. PMID- 12628174 TI - Simple networks for spike-timing-based computation, with application to olfactory processing. AB - Spike synchronization across neurons can be selective for the situation where neurons are driven at similar firing rates, a "many are equal" computation. This can be achieved in the absence of synaptic interactions between neurons, through phase locking to a common underlying oscillatory potential. Based on this principle, we instantiate an algorithm for robust odor recognition into a model network of spiking neurons whose main features are taken from known properties of biological olfactory systems. Here, recognition of odors is signaled by spike synchronization of specific subsets of "mitral cells." This synchronization is highly odor selective and invariant to a wide range of odor concentrations. It is also robust to the presence of strong distractor odors, thus allowing odor segmentation within complex olfactory scenes. Information about odors is encoded in both the identity of glomeruli activated above threshold (1 bit of information per glomerulus) and in the analog degree of activation of the glomeruli (approximately 3 bits per glomerulus). PMID- 12628172 TI - Opioid-induced quantal slowing reveals dual networks for respiratory rhythm generation. AB - Current consensus holds that a single medullary network generates respiratory rhythm in mammals. Pre-Botzinger Complex inspiratory (I) neurons, isolated in transverse slices, and preinspiratory (pre-I) neurons, found only in more intact en bloc preparations and in vivo, are each proposed as necessary for rhythm generation. Opioids slow I, but not pre-I, neuronal burst periods. In slices, opioids gradually lengthened respiratory periods, whereas in more intact preparations, periods jumped nondeterministically to integer multiples of the control period (quantal slowing). These findings suggest that opioid-induced quantal slowing results from transmission failure of rhythmic drive from pre-I neurons to preBotC I networks, depressed below threshold for spontaneous rhythmic activity. Thus, both I (in the slice), and pre-I neurons are sufficient for respiratory rhythmogenesis. PMID- 12628175 TI - Interacting roles of attention and visual salience in V4. AB - Attention increases the contrast gain of V4 neurons, causing them to respond to an attended stimulus as though its contrast had increased. When multiple stimuli appear within a neuron's receptive field (RF), the neuron responds primarily to the attended stimulus. This suggests that cortical cells may be "hard wired" to respond preferentially to the highest-contrast stimulus in their RF, and neural systems for attention capitalize on this mechanism by dynamically increasing the effective contrast of the stimulus that is task relevant. To test this, we varied the relative contrast of two stimuli within the recorded neurons' RFs, while the monkeys attended away to another location. Increasing the physical contrast of one stimulus caused V4 neurons to respond preferentially to that stimulus and reduced their responses to competing stimuli. When attention was directed to the lower-contrast stimulus, it partially overcame the influence of a competing, higher-contrast stimulus. PMID- 12628176 TI - Viewpoint-specific scene representations in human parahippocampal cortex. AB - The "parahippocampal place area" (PPA) responds more strongly in functional magnetic resonance imaging (fMRI) to scenes than to faces, objects, or other visual stimuli. We used an event-related fMRI adaptation paradigm to test whether the PPA represents scenes in a viewpoint-specific or viewpoint-invariant manner. The PPA responded just as strongly to viewpoint changes that preserved intrinsic scene geometry as it did to complete scene changes, but less strongly to object changes within the scene. In contrast, lateral occipital cortex responded more strongly to object changes than to spatial changes. These results demonstrate that scene processing in the PPA is viewpoint specific and suggest that the PPA represents the relationship between the observer and the surfaces that define local space. PMID- 12628177 TI - The well-worn route and the path less traveled: distinct neural bases of route following and wayfinding in humans. AB - Finding one's way in a large-scale environment may engage different cognitive processes than following a familiar route. The neural bases of these processes were investigated using functional MRI (fMRI). Subjects found their way in one virtual-reality town and followed a well-learned route in another. In a control condition, subjects followed a visible trail. Within subjects, accurate wayfinding activated the right posterior hippocampus. Between-subjects correlations with performance showed that good navigators (i.e., accurate wayfinders) activated the anterior hippocampus during wayfinding and head of caudate during route following. These results coincide with neurophysiological evidence for distinct response (caudate) and place (hippocampal) representations supporting navigation. We argue that the type of representation used influences both performance and concomitant fMRI activation patterns. PMID- 12628178 TI - SET-ting the stage for life and death. AB - Cytotoxic T lymphocytes release granzymes (Gzm) A and B to induce apoptosis or programmed cell death of virally infected or tumor cells. In this issue of Cell, Fan et al. identify the tumor metastasis suppressor NM23-H1 as a GzmA-activated, apoptosis-inducing DNase and the oncoprotein SET as its inhibitor. Work from the Lieberman and Wang groups indicates a surprising role for a group of acidic nucleo-cytoplasmic proteins in regulating apoptosis. PMID- 12628179 TI - The histone minority report: the variant shall not be silenced. AB - In this issue of Cell, Meneghini et al. demonstrate that replacement of canonical histone H2A with the histone variant H2A.Z within euchromatin prevents silent chromatin proteins from migrating into regions of the chromosome that normally are transcriptionally active. Thus, this highly conserved histone variant is critical for defining chromatin domains. PMID- 12628180 TI - Communication by touch: role of cellular extensions in complex animals. AB - Neurons make networks of intercellular connections. Many other cells also send out long cellular extensions and "touch" other cells far away. The extensions are intriguing, but what do they do? Possible rationales for this cell behavior include: (1) forming precise binary connections with the possibility of forming more complex networks, and (2) coupling of signaling with physical force. PMID- 12628181 TI - Polycomb, epigenomes, and control of cell identity. AB - In development, cell identity is maintained by epigenetic functions that prevent changes in cell type-specific transcription programs. Recent insights into gene silencing mechanisms by Polycomb group (PcG) and trithorax group (trxG) proteins reveal that the memory system involves a concerted process of chromatin modification, blocking of RNA polymerase II, and synthesis of noncoding RNA. Remarkably, cell memory is regulated by a balance between repressors and activators that maintains both transcription status and at the same time the possibility of switching to a different state. PMID- 12628182 TI - Functional expression of murine V2R pheromone receptors involves selective association with the M10 and M1 families of MHC class Ib molecules. AB - The vomeronasal organ (VNO) of the mouse has two neuronal compartments expressing distinct families of pheromone receptors, the V1Rs and the V2Rs. We report here that two families of major histocompatibility complex (MHC) class Ib molecules, the M10 and the M1 families, show restricted expression in V2R-expressing neurons. Our data suggest that neurons expressing a given V2R specifically co express one or a few members of the M10 family. Biochemical and immunocytochemical analysis demonstrates that in VNO sensory dendrites M10s belong to large multi-molecular complexes that include pheromone receptors and beta2-microglobulin (beta2m). In cultured cells, M10s appear to function as escort molecules in transport of V2Rs to the cell surface. Accordingly, beta2m deficient mice exhibit mislocalization of V2Rs in the VNO and a specific defect in male-male aggressive behavior. The functional characterization of M10 highlights an unexpected role for MHC molecules in pheromone detection by mammalian VNO neurons. PMID- 12628183 TI - The immunoglobulin superfamily protein SYG-1 determines the location of specific synapses in C. elegans. AB - During nervous system development, neurons form reproducible synapses onto specific targets. Here, we analyze the development of stereotyped synapses of the C. elegans HSNL neuron in vivo. Postsynaptic neurons and muscles were not required for accurate synaptic vesicle clustering in HSNL. Instead, vulval epithelial cells that contact HSNL act as synaptic guidepost cells that direct HSNL presynaptic vesicles to adjacent regions. The mutant syg-1(ky652) has defects in synapse formation that resemble those in animals that lack vulval epithelial cells: HSNL synaptic vesicles fail to accumulate at normal synaptic locations and form ectopic anterior clusters. syg-1 encodes an immunoglobulin superfamily protein that acts in the presynaptic HSNL axon. SYG-1 protein is localized to the site of future synapses, where it initiates synapse formation and localizes synaptic connections in response to the epithelial signal. SYG-1 is related to Drosophila IrreC and vertebrate NEPH1 proteins, which mediate cell cell recognition in diverse developmental contexts. PMID- 12628184 TI - Phosphorylation of the AMPA receptor GluR1 subunit is required for synaptic plasticity and retention of spatial memory. AB - Plasticity of the nervous system is dependent on mechanisms that regulate the strength of synaptic transmission. Excitatory synapses in the brain undergo long term potentiation (LTP) and long-term depression (LTD), cellular models of learning and memory. Protein phosphorylation is required for the induction of many forms of synaptic plasticity, including LTP and LTD. However, the critical kinase substrates that mediate plasticity have not been identified. We previously reported that phosphorylation of the GluR1 subunit of AMPA receptors, which mediate rapid excitatory transmission in the brain, is modulated during LTP and LTD. To test if GluR1 phosphorylation is necessary for plasticity and learning and memory, we generated mice with knockin mutations in the GluR1 phosphorylation sites. The phosphomutant mice show deficits in LTD and LTP and have memory defects in spatial learning tasks. These results demonstrate that phosphorylation of GluR1 is critical for LTD and LTP expression and the retention of memories. PMID- 12628185 TI - HIF-1alpha is essential for myeloid cell-mediated inflammation. AB - Granulocytes and monocytes/macrophages of the myeloid lineage are the chief cellular agents of innate immunity. Here, we have examined the inflammatory response in mice with conditional knockouts of the hypoxia responsive transcription factor HIF-1alpha, its negative regulator VHL, and a known downstream target, VEGF. We find that activation of HIF-1alpha is essential for myeloid cell infiltration and activation in vivo through a mechanism independent of VEGF. Loss of VHL leads to a large increase in acute inflammatory responses. Our results show that HIF-1alpha is essential for the regulation of glycolytic capacity in myeloid cells: when HIF-1alpha is absent, the cellular ATP pool is drastically reduced. The metabolic defect results in profound impairment of myeloid cell aggregation, motility, invasiveness, and bacterial killing. This role for HIF-1alpha demonstrates its direct regulation of survival and function in the inflammatory microenvironment. PMID- 12628186 TI - Tumor suppressor NM23-H1 is a granzyme A-activated DNase during CTL-mediated apoptosis, and the nucleosome assembly protein SET is its inhibitor. AB - Granzyme A (GzmA) induces a caspase-independent cell death pathway characterized by single-stranded DNA nicks and other features of apoptosis. A GzmA-activated DNase (GAAD) is in an ER associated complex containing pp32 and the GzmA substrates SET, HMG-2, and Ape1. We show that GAAD is NM23-H1, a nucleoside diphosphate kinase implicated in suppression of tumor metastasis, and its specific inhibitor (IGAAD) is SET. NM23-H1 binds to SET and is released from inhibition by GzmA cleavage of SET. After GzmA loading or CTL attack, SET and NM23-H1 translocate to the nucleus and SET is degraded, allowing NM23-H1 to nick chromosomal DNA. GzmA-treated cells with silenced NM23-H1 expression are resistant to GzmA-mediated DNA damage and cytolysis, while cells overexpressing NM23-H1 are more sensitive. PMID- 12628187 TI - DOCK4, a GTPase activator, is disrupted during tumorigenesis. AB - We used representational difference analysis to identify homozygous genomic deletions selected during tumor progression in the mouse NF2 and TP53 tumor model. We describe a deletion targeting DOCK4, a member of the CDM gene family encoding regulators of small GTPases. DOCK4 specifically activates Rap GTPase, enhancing the formation of adherens junctions. DOCK4 mutations are present in a subset of human cancer cell lines; a recurrent missense mutant identified in human prostate and ovarian cancers encodes a protein that is defective in Rap1 activation. The engulfment defect of C. elegans mutants lacking the CDM gene ced 5 is rescued by wild-type DOCK4, but not by the mutant allele. Expression of wild type, but not mutant, DOCK4 in mouse osteosarcoma cells with a deletion of the endogenous gene suppresses growth in soft agar and tumor invasion in vivo. DOCK4 therefore encodes a CDM family member that regulates intercellular junctions and is disrupted during tumorigenesis. PMID- 12628188 TI - Structural evidence for feedback activation by Ras.GTP of the Ras-specific nucleotide exchange factor SOS. AB - Growth factor receptors activate Ras by recruiting the nucleotide exchange factor son of sevenless (SOS) to the cell membrane, thereby triggering the production of GTP-loaded Ras. Crystallographic analyses of Ras bound to the catalytic module of SOS have led to the unexpected discovery of a highly conserved Ras binding site on SOS that is located distal to the active site and is specific for Ras.GTP. The crystal structures suggest that Ras.GTP stabilizes the active site of SOS allosterically, and we show that Ras.GTP forms ternary complexes with SOS(cat) in solution and increases significantly the rate of SOS(cat)-stimulated nucleotide release from Ras. These results demonstrate the existence of a positive feedback mechanism for the spatial and temporal regulation of Ras. PMID- 12628189 TI - Exit from exit: resetting the cell cycle through Amn1 inhibition of G protein signaling. AB - In S. cerevisiae cells undergoing anaphase, a ras-related GTPase, Tem1, is located on the spindle pole body that enters the daughter cell and activates a signal transduction pathway, MEN, to allow mitotic exit. MEN activation must be reversed after mitotic exit to reset the cell cycle in G1. We find that daughter cells activate an Antagonist of MEN pathway (AMEN) in part through induction of the Amn1 protein that binds directly to Tem1 and prevents its association with its target kinase Cdc15. Failure of Amn1 function results in defects of both the spindle assembly and nuclear orientation checkpoints and delays turning off Cdc14 in G1. Thus, Amn1 is part of a daughter-specific switch that helps cells exit from mitotic exit and reset the cell cycle. PMID- 12628190 TI - Structure of the catalytic domain of human DOT1L, a non-SET domain nucleosomal histone methyltransferase. AB - Dot1 is an evolutionarily conserved histone methyltransferase that methylates lysine-79 of histone H3 in the core domain. Unlike other histone methyltransferases, Dot1 does not contain a SET domain, and it specifically methylates nucleosomal histone H3. We have solved a 2.5 A resolution structure of the catalytic domain of human Dot1, hDOT1L, in complex with S-adenosyl-L methionine (SAM). The structure reveals a unique organization of a mainly alpha helical N-terminal domain and a central open alpha/beta structure, an active site consisting of a SAM binding pocket, and a potential lysine binding channel. We also show that a flexible, positively charged region at the C terminus of the catalytic domain is critical for nucleosome binding and enzymatic activity. These structural and biochemical analyses, combined with molecular modeling, provide mechanistic insights into the catalytic mechanism and nucleosomal specificity of Dot1 proteins. PMID- 12628191 TI - Conserved histone variant H2A.Z protects euchromatin from the ectopic spread of silent heterochromatin. AB - Boundary elements hinder the spread of heterochromatin, yet these sites do not fully account for the preservation of adjacent euchromatin. Histone variant H2A.Z (Htz1 in yeast) replaces conventional H2A in many nucleosomes. Microarray analysis revealed that HTZ1-activated genes cluster near telomeres. The reduced expression of most of these genes in htz1Delta cells was reversed by the deletion of SIR2 (sir2Delta) suggesting that H2A.Z antagonizes telomeric silencing. Other Htz1-activated genes flank the silent HMR mating-type locus. Their requirement for Htz1 can be bypassed by sir2Delta or by a deletion encompassing the silencing nucleation sites in HMR. In htz1Delta cells, Sir2 and Sir3 spread into flanking euchromatic regions, producing changes in histone H4 acetylation and H3 4 methylation indicative of ectopic heterochromatin formation. Htz1 is enriched in these euchromatic regions and acts synergistically with a boundary element to prevent the spread of heterochromatin. Thus, euchromatin and heterochromatin each contains components that antagonize switching to the opposite chromatin state. PMID- 12628192 TI - Bioaccumulation of trace metals in the Antarctic amphipod Orchomene plebs: evaluation of toxicokinetic models. AB - Bioaccumulation of Cd, Co, Cu, Ni, Pb and Zn in Antarctic gammaridean amphipod collectives, Orchomene plebs (Hurley, 1965), was investigated during a cruise of RV "Polarstern" to the Wedell Sea. With the sole exception of Cd the organisms accumulated metals during exposure and depurated them in uncontaminated seawater. Four independent toxicokinetic experiments and one field study were modelled simultaneously to calculate the following size-dependent bioconcentration factors for organisms with body length 10 mm (BCF(10 mm)): 130 (Co), 4030 (Cu), 190 (Ni), 2900 (Pb), and 5210 (Zn). On the time scale of our experiments the data suggest an increased metal uptake by previously exposed test organisms. The collectives investigated may be regarded as potentially suitable biomonitors for Co, Cu, Ni, Pb and Zn but not for Cd. An approach to evaluate the sensitivity of Orchomene plebs as a biomonitor of waterborne metals in the field indicates minimal increments of the ambient exposure concentrations of 0.01 microg Co l(-1), 0.2 microg Cu l(-1), 0.4 microg Ni l(-1), 0.6 microg Pb l(-1) and 0.3 microg Zn l( 1). PMID- 12628193 TI - Water quality in Sepetiba Bay, Brazil. AB - Sepetiba Bay is located at 23 degrees S, 44 degrees W in Rio de Janeiro State, Brazil. Water samples were taken at eight locations adjacent to the north shore of the Bay, near to villages and towns without sewage treatment provision. The samples were analysed and total and faecal coliform concentrations determined. A hydrodynamic model of the Bay was used together with a species dispersion model based on an adaptive quadtree mesh to predict faecal concentrations in the Bay. Effluent sources used in the model were defined using population data from census returns with flow and concentration values estimated using standard values recommended by the World Bank (WB) and the World Health Organisation (WHO). Sufficient agreement was obtained between the measured and predicted concentrations to support the use of WB and WHO summary statistics to estimate sources of sewage. PMID- 12628194 TI - Responses of a Mediterranean soft bottom community to short-term (1993-1996) hydrological changes in the Rhone river. AB - The polychaete fauna of muddy bottoms off the Rhone delta (NW Mediterranean) was seasonally sampled at two sites at 70 m depth, from 1993 to 1996. During this period, five severe flood events occurred. A clustering analysis (distance coefficient of Whittaker) and three way fixed factor ANOVAs (site x season x year) showed strong year-to-year changes in species density and community structure, changes that masked seasonal variations. Total density increased by a factor of 3 and density of most species significantly increased following the first flooding event. Changes in the community structure were due to the successive dominance of a few species. The opportunistic species, such as Cossura sp., Mediomastus sp. and Polycirrus sp., exhibited peaks in density 1-3 months after the flood. These peaks were followed by a drastic decline. For species with a long life span, such as Sternaspis scutata, a slower but continuous increase in density was observed which was maintained for several months. The density fluctuations of these species exhibited a good correlation with river flow with time lags of 1-2 years. The successional dynamics observed are explained according to the geographical origin of the floods and the biology and feeding ecology of species. PMID- 12628196 TI - Continuing effects of the Exxon Valdez oil spill. PMID- 12628195 TI - Implications of spatial heterogeneity for management of marine protected areas (MPAs): examples from assemblages of rocky coasts in the northwest Mediterranean. AB - Marine protected areas (MPAs) are increasingly used as a management tool to preserve species and habitats. Testing hypotheses about the effectiveness of MPAs is important for their implementation and to identify informative criteria to support management decisions. This study tested the general proposition that MPAs affected assemblages of algae and invertebrates between 0.0 and 0.5 m above the mean low water level of rocky coasts on two islands in the Tuscan Archipelago (northwest Mediterranean). Protection was concentrated mainly on the west coasts of the islands, raising the possibility that neither the full range of assemblages nor the relevant scales of variation were properly represented within MPAs. This motivated the comparison of assemblages on opposite sides of islands (habitats). The effects of MPAs and habitat were assessed with a multifactorial sampling design; hypotheses were tested about differences in structure of assemblages, in mean abundance of common taxa and in univariate and multivariate measures of spatial variation. The design consisted of three replicate shores for each condition of protected and reference areas on the west side of each island and three unprotected shores on the eastern side. Assemblages were sampled independently four times on each island between June 1999 and January 2001. At each time of sampling two sites were selected randomly at each of two tidal heights to represent midshore and lowshore assemblages on each shore. Estimates of abundance were obtained using non-destructive sampling methods from five replicate 20x20 cm quadrats at each site. Results indicated differences among habitats in structure of assemblages, in mean abundance of common taxa and in univariate and multivariate measures of spatial variation at the scale of shores. Most of these patterns were inconsistent with the predicted effect of management through MPAs. The data suggest that designation of MPAs in the Tuscan Archipelago should proceed through management of multiple shores and types of habitat selected to guarantee protection to a representative sample of assemblages and to the processes responsible for maintenance of spatial patchiness at different scales. This study also shows that considerations of spatial heterogeneity are important to underpin management decisions about the number, size and location of MPAs. PMID- 12628197 TI - The behaviour of residual contaminants at a former station site, Antarctica. AB - In 1994, New Zealand's only mainland Antarctic base, Vanda Station, was removed from the shores of Lake Vanda, in the McMurdo Dry Valleys region of southern Victoria Land, Antarctica. Residual chemical contamination of the station site has been identified, in the form of discrete fuel spills, locally elevated Pb, Zn, Ag and Cd concentrations in soil and elevated Cu, Ni, Co and phosphate concentrations in suprapermafrost fluids in a gully formerly used for domestic washing water disposal. Pathways for contaminant transfer to Lake Vanda, potential environmental impacts and specific remediation/monitoring options are considered. While some contaminants (particularly Zn) could be selectively leached from flooded soil, during a period of rising lake level, the small area of contaminated soils exposed and low level of contamination suggests that this would not adversely affect either shallow lake water quality or the growth of cyanobacteria. Phosphate-enhanced growth of the latte may, however, be a visible consequence of the minor contamination occurring at this site. PMID- 12628198 TI - Total and methyl mercury concentrations and fluxes from small boreal forest catchments in Finland. AB - Total mercury (TotHg) and methyl mercury (MeHg) concentrations were studied in runoff from eight small (0.02-1.3 km2) boreal forest catchments (mineral soil and peatland) during 1990-1995. Runoff waters were extremely humic (TOC 7-70 mg l-1). TotHg concentrations varied between 0.84 and 24 ng l-1 and MeHg between 0.03 and 3.8 ng l-1. TotHg fluxes from catchments ranged from 0.92 to 1.8 g km-2 a-1, and MeHg fluxes from 0.03 to 0.33 g km-2 a-1. TotHg concentrations and output fluxes measured in runoff water from small forest catchments in Finland were comparable with those measured in other boreal regions. By contrast, MeHg concentrations were generally higher. Estimates for MeHg output fluxes in this study were comparable at sites with forests and wetlands in Sweden and North America, but clearly higher than those measured at upland or agricultural sites in other studies. Peatland catchments released more MeHg than pure mineral soil or mineral soil catchments with minor area of peatland. PMID- 12628199 TI - Functional and structural rearrangements of salivary gland polytene chromosomes of Chironomus riparius Mg. (Diptera,Chironomidae) in response to freshly neutralized aluminium. AB - Although recent work has shown that environmentally relevant concentrations of freshly neutralized aluminium (AI) are bioavailable and toxic to freshwater invertebrates, the genotoxicity of Al has not been examined. Here we show that freshly neutralized Al affects structure and function of the salivary gland polytene chromosomes of the ubiquitous chironomid larva Chironomus riparius over three generations. Exposure to 500 microg l-1 added Al for 24-25 days resulted in a significantly higher frequency of numerous somatic aberrations, while no structural aberrations were found in F1 controls and few in the second and third generation. Aberrations also included deletions of sections of chromosome G of C. riparius larvae as well as deletions of one or more Balbiani rings. Changes in functional activity included decreased activity of the Balbiani rings (BR), and an increase in the number of decondensed centromeres. The activity of the nucleolar organizer (NOR) significantly decreased in F1 chironomids exposed to Al, while in the F2 and F3 generations the NOR showed normal (high) activity. First generation chironomids were generally more susceptible to Al although no clear evidence of tolerance was apparent over three generations. The possible use of alterations in chironomid polytene chromosomes as biomarkers of trace metal pollution is discussed. PMID- 12628200 TI - Ozone differentially affects physiological and biochemical responses of two clover species; Trifolium repens and Trifolium pratense. AB - The effect of acute ozone exposure (150 ppb for 3 h) on two clover species, white clover (Trifolium repens L.) and red clover (Trifolium pratense L.) was investigated through the analysis of 10 different physiological and biochemical parameters. Twenty-four hours after fumigation, visible symptoms of injury on leaves were observed only in red clover, but from the biochemical point of view, both species revealed significant ozone-induced modifications. A decrease in the photosynthetic efficiency as well as an increase in the de-epoxidation index and a decrease in the redox state of ascorbate were detected only in T. pratense leaves; no significant change in pigment content was found in either of the two species. On the other hand, both white and red clover showed, although to different extents, significant decreases in the activities of the antioxidant enzymes peroxidase and ascorbate peroxidase. Multivariate statistical analysis revealed not only that ozone affects both species, but also that they differentially respond to the pollutant, confirming the higher sensitivity of Trifolium pratense to ozone exposure. PMID- 12628201 TI - Assessment of adsorption behavior of dibutyltin (DBT) to clay-rich sediments in comparison to the highly toxic tributyltin (TBT). AB - The sorption behavior of dibutyltin (DBT) to four types of natural clay-rich sediments as a function of pH and salinity was studied. The strongest affinity of DBT was found to the montmorillonite-rich sediment, which is characterized by the highest specific surface area and cation exchange capacity of the four used sediments. Kd values range between 12 and 40 (l/kg) on simulated marine conditions (pH 8, salinity 32%). A maximum of DBT adsorption was found at a salinity of 0% and pH 6. Desorption occurred over the entire studied pH range (4 8) when contaminated sediments interact with butyltin-free water. The maximum of desorption coincided with the minimum of adsorption, and vice versa. The results of DBT adsorption are compared with tributyltin (TBT), and the mechanism of the adsorption process is discussed. PMID- 12628202 TI - Copper bioavailability and extractability as related to chemical properties of contaminated soils from a vine-growing area. AB - Vineyard soils have been contaminated by Cu as a consequence of the long-term use of Cu salts as fungicides against mildew. This work aimed at identifying which soil parameters were the best related to Cu bioavailability, as assessed by measuring the concentrations of Cu in shoots and roots of tomato cropped (in lab conditions) over a range of 29 (24 calcareous and five acidic) Cu-contaminated topsoils from a vine-growing area (22-398 mg Cu kg(-1)). Copper concentrations in tomato shoots remained in the adequate range and were independent of soil properties and soil Cu content. Conversely, strong, positive correlations were found between root Cu concentration, total soil Cu, EDTA- or K-pyrophosphate extractable Cu and organic C contents in the 24 calcareous soils, suggesting a prominent role of organic matter in the retention and bioavailability of Cu. Such relations were not observed when including the five acidic soils in the investigated population, suggesting a major pH effect. Root Cu concentration appeared as a much more sensitive indicator of soil Cu bioavailability than shoot Cu concentration. Simple extractions routinely used in soil testing procedures (total and EDTA-extractable Cu) were adequate indicators of Cu bioavailability for the investigated calcareous soils, but not when different soil types were considered (e.g. acidic versus calcareous soils). PMID- 12628204 TI - Composition and element solubility of magnetic and non-magnetic fly ash fractions. AB - Magnetic and non-magnetic fractions of coal fly ashes from SE US electric power plants were characterized with special emphasis on the potential environmental consequences of their terrestrial disposal. Quartz and mullite were the crystalline minerals dominating the non-magnetic fractions. Magnetic fractions contained magnetite, hematite, and, to a lesser extent, quartz and mullite. Chemical analyses revealed that magnetic fractions had about 10 times higher concentrations of Fe, and 2-4 times higher concentrations of Co, Ni, and Mn. Non magnetic fractions were enriched in K, Al and Ca. Iron content within fly ash particles was negatively correlated with elements associated with aluminosilicate matrix (Si, Al, K, Na). Solubility of most elements was higher in the non magnetic than in the magnetic fractions of alkaline fly ashes at comparable pH. Calcium was associated with the non-magnetic fraction of the alkaline fly ashes which resulted in a higher pH buffering capacity of this fraction. PMID- 12628203 TI - Predicting the release of metals from ombrotrophic peat due to drought-induced acidification. AB - Ombrotrophic peats in northern England and Scotland, close to industrial areas, have substantial contents of potentially toxic metals (Al, Ni, Cu, Zn, Cd and Pb) and of pollutant sulphur, all derived from atmospheric deposition. The peat sulphur, ordinarily in reduced form, may be converted to sulphuric acid under drought conditions, due to the entry of oxygen into the peats. The consequent lowering of soil solution pH is predicted to cause the release of metals held on ligand sites of the peat organic matter. The purpose of the present study was to explore, by simulation modelling, the extent of the metal response. Chemical variables (elemental composition, pH, metal contents) were measured for samples of ombrotrophic peats from three locations. Water extracts of the peats, and samples of local surface water, were also analysed, for pH, dissolved organic carbon (DOC) and metals. Metal release from peats due to acidification was demonstrated experimentally, and could be accounted for reasonably well using a speciation code (WHAM/Model VI). These data, together with information on metal and S deposition, and meteorology, were used to construct a simple description of peat hydrochemistry, based on WHAM/Model VI, that takes into account ion-binding by humic substances (assumed to be the "active" constituents of the peat with respect to ion-binding). The model was used to simulate steady state situations that approximated the observed soil pH, metal pools and dissolved metal concentrations. Then, drought conditions were imposed, to generate increased concentrations of H2SO4, in line with those observed during the drought of 1995. The model calculations suggest that the pH will decrease from the initial steady state value of 4.3 to 3.3-3.6 during rewetting periods following droughts, depending upon assumptions about the amount of potentially mobile soil S. The pH decreases will be accompanied by increases in concentrations of dissolved metals (Mg, Al, Ca, Ni, Cu, Zn, Cd, Pb) of an order of magnitude or more, depending upon assumptions about the replenishment of soil metal pools by deposition. In the most realistic scenario for present conditions, the severity of pH depressions will gradually decline due to the relatively slow depletion of the soil S pool by droughts. However, the magnitudes of heavy metal pulses will decline quite rapidly (over two or three droughts) because current and future metal deposition is unable to compensate for leaching losses from the soil pools. PMID- 12628205 TI - Effects of phytosterols on zebrafish reproduction in multigeneration test. AB - Zebrafish from mixed sex populations were exposed continuously across three generations to two phytosterol preparations both containing -sitosterol. The phytosterols were isolated from wood and soy beans. Blood vitellogenin levels and sex ratio changes were used as intermediate indicators of the reproduction failures. Both sterol preparations caused vitellogenin induction in the exposed fish. The wood sterol changed the sex ratio of the exposed fish. In generation F1, the predominant sex was male, and in generation F2 it was female. The soy sterol in the used test concentration was lethal to the exposed fish in generation F1. This multigeneration test evidenced that phytosterols containing sitosterol disrupt the reproduction system of zebrafish by changing the sex ratios and by inducing the vitellogenin production in the exposed fish. PMID- 12628206 TI - Monitoring the effects of atmospheric ethylene near polyethylene manufacturing plants with two sensitive plant species. AB - Data of a multi-year (1977-1983) biomonitoring programme with marigold and petunia around polyethylene manufacturing plants was analysed to assess plant responses to atmospheric ethylene and to determine the area at risk for the phytotoxic effects of this pollutant. In both species, flower formation and growth were severely reduced close to the emission sources and plant performance improved with increasing distance. Plants exposed near the border of the research area had more flowers than the unexposed control while their growth was normal. Measurements of ethylene concentrations at a border site revealed that the growing season mean was 61.5 g m(-3) in 1982 and 15.6 g m(-3) in 1983. In terms of number of flowers, petunia was more sensitive than marigold and adverse effects were observed within ca. 400 m distance from the sources for marigold and within ca. 460 m for petunia. The area at risk (ca. 870 m) for ethylene-induced growth reduction was also limited to the industrial zone. Plants were more sensitive to ethylene in terms of growth reduction than in terms of inhibition of flowering. In the Netherlands, maximum permissible levels of ethylene are currently based on information from laboratory and greenhouse studies. Our results indicate that these levels are rather conservative in protecting field grown plants against ethylene-induced injury near polyethylene manufacturing plants. PMID- 12628207 TI - Correlation between the nitrogen concentration of two epiphytic lichens and the traffic density in an urban area. AB - A field experiment was carried out in the urban environment of the Grenoble area using two epiphytic lichens: the nitrophytic Physcia adscendens and the acidiphytic Hypogymnia physodes. Two complementary studies characterized this experiment. Firstly, a sampling of the two lichens in 48 sites randomly located throughout the Grenoble area indicated that roads (size and proximity to sampling sites) influenced the nitrogen concentrations of P. adscendens, but not those of H. physodes. Secondly, to study more accurately the influence of roads, a traffic index was calculated and applied along two transects located perpendicularly to urban motorways. Significant positive correlations were found between this traffic index and the total nitrogen concentration of P. adscendens. PMID- 12628208 TI - Organic compounds and trace metals of anthropogenic origin in sediments from Montego Bay, Jamaica: assessment of sources and distribution pathways. AB - Surface sediments throughout Montego Bay, Jamaica were collected in 1995 and analyzed for their trace metal and trace organic contaminant content. A variety of trace metals, petroleum hydrocarbons, polycyclic aromatic hydrocarbons, coprostanol as well as chlorinated hydrocarbons such as pesticides and polychlorinated biphenyls were detected and provide evidence for several anthropogenic inputs to the bay. Two main sources of these chemicals are the Montego River and the North Gully, the latter being more significant. Particle associated pollutants were found to be distributed along the Montego River plume, as well as being transported by the prevailing water currents to the South Western sections of the bay, probably through re-suspension of enriched fine sediments from the North Gully outfall area. PMID- 12628210 TI - On the local and regional influence on ground-level ozone concentrations in Hong Kong. AB - Hong Kong is a densely populated city situated in the fast developing Pearl River Delta of southern China. In this study, the recent data on ozone (O3) and related air pollutants obtained at three sites in Hong Kong are analyzed to show the variations of O3 in urban, sub-urban and rural areas and the possible regional influences. Highest monthly averaged O3 was found at a northeastern rural site and lowest O3 level was observed at an urban site. The levels of NOx, CO, SO2 and PM10 showed a different spatial pattern with the highest level in the urban site and lowest at the rural site. Analysis of chemical species ratios such as SO2/NOx and CO/NOx indicated that the sites were under the influences of local and regional emissions to varying extents reflecting the characteristics of emission sources surround the respective sites. Seasonal pattern of O3 is examined. Low O3 level was found in summer and elevated levels occurred in autumn and spring. The latter appears different from the previous result obtained in 1996 indicating a single maximum occurring in autumn. Principal component analysis was used to further elucidate the relationships of air pollutants at each site. As expected, the O3 variation in the northeastern rural area was largely determined by regional chemical and transport processes, while the O3 variability at the southwestern suburban and urban sites were more influenced by local emissions. Despite the large difference in O3 levels across the sites, total potential ozone (O3+NO2) showed little variability. Cases of high O3 episodes were presented and elevated O3 levels were formed under the influence of tropical cyclone bringing in conditions of intense sunlight, high temperature and light winds. Elevated O3 levels were also found to correlate with enhanced ratio of SO2 to NOx, suggesting influence of regional emissions from the adjacent Pearl River Delta region. PMID- 12628209 TI - The effect of iron-ore particles on the metal content of the brown alga Padina gymnospora (Espirito Santo Bay, Brazil). AB - The iron-ore particles discharged by a pellet processing plant (Espirito Santo Bay, Brazil) cover the seabed of Camburi Beach and consequently, the epibenthic community. In order to determine the importance of the contribution of the iron ore deposits to the metal concentration in macroalgae of Espirito Santo Bay, four methods of cleaning particulate material adhered to the surface of thalli were tested prior to metal tissue analysis (Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) of Padina gymnospora. In addition, heavy metal concentrations were determined in individuals of P. gymnospora from a site (Frade Island) not affected by the iron ore particles. The most efficient cleaning treatment, a combination of scraping and washing with an ethanol-seawater solution (NA+SC+ET) removed a number of particles on the surface of thalli 10 times higher than that observed in the control (C). Using this treatment, the total-metal concentrations were reduced by 78% for Fe and 50% for Al respect to the control. However, Fe, Al and Cu concentrations after treatment NA+SC+ET were significantly higher than those found at Frade Island. It is suggested that the iron-ore deposit might be a source for metal availability to macroalgae exposed to the dumped material at Espirito Santo Bay. PMID- 12628212 TI - Sleep in medical disorders. PMID- 12628211 TI - Concentrations of heavy metals in Sotalia fluviatilis (Cetacea: Delphinidae) off the coast of Ceara, northeast Brazil. AB - Lead (Pb), cadmium (Cd) and mercury (Hg) concentrations on liver and kidney of Sotalia fluviatilis (Cetacea: Delphinidae) stranded in the coast of Ceara, Brazil, were studied from 1996 to 1999. Pb levels were usually lower than the detection limit (0.1 microg/g). Concentrations of Cd were significantly higher in kidney than liver, averaging 0.8 microg/g. Mercury accumulation took place mainly in liver with an average concentration of 4.6 microg/g. Both metals were significantly higher in larger mature individuals, but differences between sexes were not significant. The detection of Cd, Hg and Pb in tissue samples of S. fluviatilis off the coast of Ceara indicated that heavy metals are locally available in the water, and bioaccumulation may be occurring through the food web. Contamination levels were not considered critical, but could be related to Ceara's growing industrial development. The associated risks of pollution outfalls may pose a threat to marine organisms in a near future, especially for top predators such as S. fluviatilis. PMID- 12628213 TI - Parkinson's disease and sleep. AB - Sleep disorders are common in Parkinson's disease (PD), as almost two thirds of PD patients report them. From a clinical point of view, they can be classified into disorders of initiation and maintenance of sleep (DIMS), parasomnias, and excessive daytime sleepiness (EDS). Among the causes of DIMS are degenerative changes in the CNS affecting centers for sleep regulation, persistence into the night of daytime PD-related symptoms, concomitant medical or psychiatric disease, disruption of circadian rhythms, and effects of dopaminergic (and other) medication on sleep regulation. Parasomnias might further contribute to sleep disturbance, as they can be accompanied by motor desinhibition during REM sleep. Parasomnias can precede by several years the presence of daytime PD symptoms. EDS has been over the last years the focus of attention for both sleep and movement disorders specialists, due to the fact that it might predispose to traffic accidents. However, the so-called "sleep attacks" never occur without preexisting somnolence. Thus, a careful sleep history can be helpful to determine which patients are exposed to suffer them. Although EDS was initially attributed to the effects of dopaminergic medication, it seems likely that several disease-related factors might also play an important role. An adequate education of the PD patients in sleep hygiene measures and a skilled use of the medication seem necessary to prevent sleep disturbance. PMID- 12628214 TI - Sleep disturbances in dialysis patients. AB - Sleep disturbances are extremely common in dialysis patients. Subjective sleep complaints are reported in up to 80% of those surveyed and sleep apnoea syndrome, restless legs syndrome, and periodic limb movement disorder are much more prevalent than in the general population. Excessive daytime sleepiness is also an important problem. These sleep abnormalities appear to have significant negative effects on quality of life and functional health status. Although long-term studies regarding other effects on health outcomes remain to be conducted, available data also suggest that sleep disturbances may have an important impact on morbidity and mortality. Achieving a more complete understanding of the sleep problems experienced by this group is absolutely imperative if improving health outcomes is the goal. Clinicians and researchers alike face numerous challenges in this regard, especially when considering the complex clinical presentation and treatment needs typical of these patients. Therefore, the purpose of this article is to present an up-to-date review of the literature regarding sleep disturbances in dialysis patients with special emphasis on the numerous factors potentially contributing to these problems and associated clinical and research implications. PMID- 12628215 TI - Brainstem modulation of pain during sleep and waking. AB - Moderately painful stimuli applied during sleep evoke motor and neural responses indicative of arousal, but seldom cause awakening. Different reactions occur in response to acute pain stimulation across behavioral states; pain reactions are modulated by the activity of serotonergic and non-serotonergic cells in the raphe magnus (RM). Serotonergic RM cells have state-dependent discharge and may inhibit simple motor withdrawal responses during waking. ON and OFF cells are non serotonergic RM neurons thought to facilitate and inhibit pain, respectively. These cells display reciprocal spontaneous discharge patterns across the sleep wake cycle, with ON cells most active during waking and OFF cells most active during sleep. We propose that they also play an important role in modulating the alertness evoked by any brief external stimulus, either noxious or innocuous. ON cells may facilitate alertness during waking and OFF cells suppress arousals during sleep. In the presence of chronic pain, both ON and OFF cell discharge appear to increase. The increase in ON cell discharge may contribute to enhancing pain sensitivity and alertness. Future research is needed to understand why sleep is so adversely affected in chronic pain patients, whereas sleep is minimally disrupted, even by acutely painful stimuli, in humans and animals without chronic pain. PMID- 12628216 TI - Sleep in women across the life cycle from adulthood through menopause. AB - Studies of sleep across the life cycle in women have utilized both survey and polysomnographic techniques, but have tended to be of small sample size with diverse methodology. As a result, definitive conclusions about the impact of the menstrual cycle and use of oral contraceptives on sleep parameters cannot yet be made. Sleep disruption during pregnancy and postpartum is nearly universal, but effective and practical countermeasures are still needed. Longitudinal studies of sleep in the postpartum period are also lacking. Menopause is associated with insomnia due to several factors including hot flashes, mood disorders and increased sleep-disordered breathing. The use of hormone replacement therapy to treat sleep and other variables is an active area of investigation. In summary, much research is required to fully elucidate the impact of the life cycle on sleep parameters in women. PMID- 12628217 TI - The development of sleep during the first months of life. AB - The development of sleep during the first months of life is intriguing from a scientific point of view but is interesting to the clinician as well. This review aims at providing data on sleep in "low-risk" premature neonates, neonates born at term, and infants up to the age of 6 months. The various milestones in clinical and polygraphic features of sleep allow for week-to-week assessment of brain function in normal and sick children. PMID- 12628218 TI - Ineffectiveness of intermittent zolpidem. PMID- 12628220 TI - Lipid peroxidation in chronic gastritis; any influence of Helicobacter pylori? AB - In an attempt to investigate the significance of lipid peroxidation in the pathogenesis of gastritis associated with or without Helicobacter pylori infection, malonodialdehyde (MDA) levels were measured by the thiobarbiturate assay in the gastric juice of 101 patients undergoing upper GI endoscopy and correlated with histopathological findings. Elevated MDA levels were found in all patients with gastritis compared with controls. MDA levels were significantly correlated with the extent of the mucosal inflammation and with disease activity in patients with reactive gastritis. In patients with H. pylori associated gastritis MDA levels were not correlated with disease activity but rather with the degree of atrophy. In this case, MDA levels were equal or even less than in patients with reactive gastritis. MDA levels were not affected by the history of consumption of PPIs, of H(2)-blockers or of NSAIDs over the last month before the endoscopy. It is concluded that lipid peroxidation is a mechanism involved in the pathogenesis of gastritis associated or not to H. pylori infection. PMID- 12628221 TI - Anti-leukotriene drugs in the prevention and treatment of hepatorenal syndrome. AB - Hepatorenal syndrome (HRS) is a peculiar form of progressive renal failure complicating the course of cirrhosis and ascites. The renal impairment of HRS is merely functional and potentially reversible. Notwithstanding, in spite of several encouraging attempts, a satisfactory medical treatment for HRS is still expected. Several pathophysiological mechanisms are active in HRS. Arachidonate metabolism derangements are among these, and prostaglandins and thromboxane antagonists have been tried with variable outcomes. Also leukotrienes (LT) appear to be involved in HRS. Three drugs (zileuton, montelukast and zafirlukast) interfering with LT synthesis and receptor binding are currently available, but they have not yet been tried in HRS. Accordingly, the author would like to suggest physicians engaged in care of these critical patients to consider a trial with these drugs-as well as with any future innovative agent active on the arachidonate-derived metabolic pathways. PMID- 12628223 TI - Characterization of 11-dehydro-thromboxane B2 recombinant antibody obtained by phage display technology. AB - Recombinant monoclonal antibodies specific for 11-dehydro-thromboxane B(2) (11D TX) were isolated from the combinatorial libraries on a pComb3 phage-display vector using a magnetic cell sorting (MACS) system. The libraries were constructed from repertories of light and heavy-chains derived from the total RNA of 11D-TX conjugated keyhole limpet haemocyanin-immunized mice. Biotinylation of 11D-TX conjugated bovine serum albumin (BSA) was performed through free thiol groups on BSA using 1-biotinamido-4-[4'-(maleimidomethyl) cyclohexanecarboxamido] butane (Biotin-BMCC). Affinity bio-panning was performed to enrich the phage display libraries against biotinylated 11D-TX conjugated BSA with the MACS system. Results indicated that the selected anti-11D-TX Fab fragments expressed by E. coli exhibited a five-fold higher affinity for BSA conjugated 11D-TX compared to BSA alone and little specificity to other related compounds as determined by the binding assay and inhibition enzyme-linked immunosorbent assay (ELISA). This is the first report of an antibody against prostaglandin produced by phage display technology and also determination of the DNA sequence of this antibody. The MACS system was shown to be a simpler and more efficient method of panning than the conventional ELISA procedure. According to our results, we concluded that the phage display technique combined with the MACS system allowed the selection of the antibody with high affinity and some specificity. PMID- 12628222 TI - The stable analog carbocyclic TXA2 but not platelet-released TXA2 induces osteoclast-like cell formation. AB - Thromboxan A(2) (TXA(2)) is the main product of arachidonic acid metabolism in activated platelets. Platelet-released supernatants (PRS) can induce osteoclast like cell formation in murine bone marrow cultures via a cyclooxygenase (COX)/receptor activator of NF-kB-ligand (RANKL)-dependent pathway. Here we investigated a possible linkage between platelet-released TXA(2) and osteoclastogenesis. The stable analog of TXA(2), carbocyclic TXA(2) (CTXA(2)) can induce the formation of tartrate-resistant acid phosphatase positive multinucleated cells in murine bone marrow cultures via a RANKL-dependent pathway and requires the presence of stromal cells. Interestingly, the platelet-released instable TXA(2) does not account for osteoclastogenic effects as: (a) PRS-induced osteoclastogenesis in the presence of the TXA(2) receptor antagonist SQ29548; (b) inhibition of platelet TXA(2) synthesis by indomethacin and acetylsalicylic acid failed to decrease the osteoclastogenic potential of the corresponding supernatants; and (c) CTXA(2)-induced osteoclast-like cell formation independent of indomethacin and the selective COX-2 inhibitor NS398. PMID- 12628224 TI - Crosstalk between nitric oxide synthase and cyclooxygenase metabolites in the estrogenized rat uterus. AB - In the present study, we investigated the effect of nitric oxide (NO) and prostaglandins (PGs) on the production of arachidonate and L-arginine metabolites. We found that in the estrogenized rat uterus lipopolysaccharide (LPS) 5mg/kg induced NO and PGs synthesis simultaneously. The uteri were incubated with different doses of an NO donor: NP 300 and 600 microM. The results indicate that both doses of NP produce a significant increase (P<0.01) in all prostanoids evaluated. The stimulatory effect was completely reversed by the addition of 2 microg/ml of hemoglobin (Hb), an NO scavenger. However, NOS inhibitor, N(G)-L-monomethyl arginine had no effect on basal prostanoid production. We also studied NO synthesis in the presence of different PGs concentration. We found that PGF(2alpha) and PGD(2) were capable of reversing LPS stimulation on NO synthesis (P<0.05), in all the doses evaluated. On the other hand, PGE(2) 10(-10) and 10(-9)M potentated LPS effect (P<0.001). These results suggest that in the estrogenized rat uterus, the synthesis of cyclooxygenase metabolites is positively regulated by NO, while NO synthesis regulation depends on the PGs evaluated. PMID- 12628225 TI - Activity, synthesis, storage, and messenger RNA of cyclooxygenase in intrauterine tissues of guinea pigs near term and during labor. AB - Whether the reported gestation-dependent increase in cyclooxygenase activity in gestational tissues is due to an accumulation of cyclooxygenase in vivo or an increasing capacity to synthesize cyclooxygenase in vitro is unknown. In this study in guinea pigs, COX activity was estimated from the net production rates of prostaglandins E(2) and F(2alpha) in the presence of optimal substrate concentrations. Cyclooxygenase activity in amnion increased between 45 days of gestation and labor in microsomes (150-fold in relation to PGF(2alpha) production and 116-fold in relation to PGE(2) production) and in tissue explants (42-fold in relation to PGF(2alpha) production). The capacity for de novo synthesis of cyclooxygenase after aspirin treatment increased nine-fold between 45 days of gestation and labor in amnion explants. Comparison of COX activity in amnion explants with or without prior aspirin treatment showed that COX activity is at least three-fold higher in controls than would be expected if the activity was due to de novo synthesis alone. Cyclooxygenase-2 mRNA predominated in amnion but neither cyclooxygenase-2 nor cyclooxygenase-1 mRNA levels (semi-quantitative RT PCR) changed significantly. This suggests that the gestation-dependent increase in cyclooxygenase activity in guinea pig amnion is due in part to accumulation of cyclooxygenase in vivo, that COX-2 predominates, and that COX activity is not correlated with levels of COX mRNA. PMID- 12628227 TI - A fully coupled binary biochemical reactive-diffusion model with analytic solution. AB - Coupled multicomponent biochemical reactive diffusion underlies a variety of biological signalling processes and pharmacokinetic applications, such as paracrine signalling involving "cocktails" comprised of growth promoter/inhibitor factors and proteases associated with tumor angiogenesis, invasion and metastasis, extravascular drug delivery, and polymeric controlled-release drug codelivery design. Here, we present a model and develop a new analytic solution to illustrate the spatiotemporal behavior associated with fully coupled binary biochemical reactive diffusion. The complete coupling renders the solution appreciably more complex in structure and behavior than solutions for unicomponent or partially coupled models. Concentration behavior is illustrated by the computational simulation of binary-species tumor angiogenesis factor reactive-diffusion in the extravascular tissue matrix. The computational results indicate that (a) steady-state concentration profiles are achieved within 1 h of a change in factor production; (b) in the steady state, the spatial profiles of the two components tend to be similar; (c) exceedingly steep concentration gradients, involving several orders-of-magnitude differences in concentration over a few tenths of a millimeter, can occur in the vicinity of boundary sources due to inter-species reaction; (d) the concentration profiles of the two species differ from unicomponent predictions due to the simultaneous mass interchange between the two species. The analytic solution predictions are also used to provide a first-ever validation of a time-dependent, binary-component Crank Nicholson numerical solution. The ability to quantitatively model interacting and often strongly varying concentration levels as a function of time and position can serve as a powerful complementary tool to experimental analyses for assessing disease state and interventional pharmacological efficacy, especially when the spatial scales on which in vivo behavior occurs taxes the limits of imaging capabilities. PMID- 12628228 TI - The role of variability and risk on the persistence of shared-enemy, predator prey assemblages. AB - The role of indirect effects such as apparent competition in structuring predator prey assemblages has recently received empirical attention. That one prey species can be excluded by the impact of a shared-enemy contrasts with the known diversity of multispecies predator-prey interactions. Here, the role of predator foraging among patches of two different prey species is examined as a mechanism that can mediate coexistence in multispecies prey-predator assemblages. Specifically, models of host-parasitoid interactions are constructed to analyse how different types of aggregative behaviour (generated by host-dependent and host-independent responses) affect persistence of the assemblage. How the distribution of hosts and the response of the parasitoid to these distributions can influence coexistence is shown. A generic explanation for coexistence suggests that it is the variability rather than the precise functional relationship that is critical for coexistence under shared-enemy interactions. PMID- 12628229 TI - Modeling growth of a heterogeneous tumor. AB - It has long been recognized that the growth of tumor population depends on the initial age distribution of the cells in the tumor and the age-dependent cellular birth rate. Deterministic dual-cell models have been available for sometime; these models take into account the effects of the resultant cell heterogeneity. Nevertheless, these models ignore various variables significantly affecting the growth, such as those characterizing the cells' inherent properties and environmental factors. Uncertainties, or fluctuations, arise when the growth is simulated with the models. Stochastic analysis of these fluctuations is the focus of the current work. Two types of cells are visualized to proliferate separately and to transform mutually during the process. The master equations of the system have been formulated through probabilistic population balance around a particular state by considering all mutually exclusive events. The governing equations for the means, variances, and covariance of the random variables have been derived through the system-size expansion of these nonlinear master equations. The stochastic pathways of the two different types of cells have been numerically simulated by the algorithm derived from the master equation for two different physical situations, one without and, the other, with the chemotherapeutic treatment. The results of the current study illuminate the significance of stochastically modeling the responses of the tumor to a variety of medicinal treatments: The coefficient of variation of the malignant cells' population magnifies with time under chemotherapeutic regimens. Consequently, the impact of the uncertainties in the exact number of malignant cells as expressed by this coefficient of variation is highly unpredictable. For example, it becomes increasingly uncertain if or how fast these cells will reactivate to become a full-blown carcinogenic tumor after treatment. PMID- 12628230 TI - Why does a cleavage plane develop parallel to the spindle axis in conical sand dollar eggs? A key question for clarifying the mechanism of contractile ring positioning. AB - Three types of models have been proposed about how the mitotic apparatus determines the position of the cleavage furrow in animal cells. In the first and second types, the contractile ring appears in a cortical region that least and most astral microtubules reach, respectively. The third type is that the spindle midzone positions the contractile ring. In the previous study, a new model was proposed through analyses of cytokinesis in sand dollar and sea urchin eggs. Gradients of the surface density of microtubule plus ends are assumed to drive membrane proteins whose accumulation causes the formation of contractile-ring microfilaments. In the present study, the validity of each model is examined by simulating the furrow formation in conical sand dollar eggs with the mitotic apparatus oriented perpendicular to the cone axis. The new model predicts that unilateral furrows with cleavage planes roughly parallel to the spindle axis appear between the mitotic apparatus and the vertex besides the normally positioned furrow. The predictions are consistent with the observations by Rappaport & Rappaport (1994, Dev. Biol.164, 258-266). The other three types of models do not predict the formation of the ectopic furrows. Furthermore, it is pointed out that only the new model has the ability to explain the geometrical relationship between the mitotic apparatus and the contractile ring under various experimental conditions. These results strongly suggest the real existence of the membrane proteins postulated in the model. PMID- 12628231 TI - Responses of Ca2+-binding proteins to localized, transient changes in intracellular [Ca2+]. AB - In smooth muscle cells, various transient, localized [Ca(2+)] changes have been observed that are thought to regulate cell function without necessarily inducing contraction. Although a great deal of effort has been put into detecting these transients and elucidating the mechanisms involved in their generation, the extent to which these transient Ca(2+) signals interact with intracellular Ca(2+) binding molecules remains relatively unknown. To understand how the spatial and temporal characteristics of an intracellular Ca(2+) signal influence its interaction with Ca(2+)-binding proteins, mathematical models of Ca(2+) diffusion and regulation in smooth muscle cells were used to study Ca(2+) binding to prototypical proteins with one or two Ca(2+)-binding sites. Simulations with the models: (1) demonstrate the extent to which the rate constants for Ca(2+)-binding to proteins and the spatial and temporal characteristics of different Ca(2+) transients influence the magnitude and time course of the responses of these proteins to the transients; (2) predict significant differences in the responses of proteins with one or two Ca(2+)-binding sites to individual Ca(2+) transients and to trains of transients; (3) demonstrate how the kinetic characteristics determine the fidelity with which the responses of Ca(2+)-sensitive molecules reflect the magnitude and time course of transient Ca(2+) signals. Overall, this work demonstrates the clear need for complete information about the kinetics of Ca(2+) binding for determining how well Ca(2+)-binding molecules respond to different types of Ca(2+) signals. These results have important implications when considering the possible modulation of Ca(2+)- and Ca(2+)/calmodulin-dependent proteins by localized intracellular Ca(2+) transients in smooth muscle cells and, more generally, in other cell types. PMID- 12628232 TI - Size scaling of mutation avalanches in a model for protein evolution. AB - The tunably rugged NK-model is used to study avalanche-like events that occur when environmental change causes fitness optima to disappear. The probability of an event with Delta substitutions scales as exp(-c Delta) for smooth landscapes, and as exp(-c Delta(2)) for rugged landscapes. Increasing the ruggedness leads to two competing effects: (1) more possible routes by which single mutations can increase the fitness, which dominates at low ruggedness and acts to increase Delta; and (2) a higher density of fitness optima, which dominates at high ruggedness and acts to decrease Delta. Due to these competing effects, the largest average values of Delta occur at intermediate ruggedness. The effects of system size on the avalanche events are examined, and average values of Delta increase logarithmically with system size. The variance to mean ratios for the number of substitutions per unit time are consistent with experimental results for protein evolution. PMID- 12628233 TI - A Kinetic Analysis of the Mechanism of beta-Amyloid Induced G Protein Activation. AB - beta-Amyloid (A beta) is the primary protein component of senile plaques found in Alzheimer's disease. In an aggregated (amyloid fibril, protofibril, or low molecular weight oligomer) state, A beta has been consistently shown to be toxic to neurons, but the molecular mechanism of this toxicity is poorly understood. We have previously shown that A beta activates a G(i/o) protein, and that inhibition of this specific G protein activation attenuated A beta-induced cell toxicity. In the present study, we use a kinetic analysis to examine the mechanism of A beta induced G protein activation. Using synthetic A beta(1-40) and phospholipid vesicles containing purified G(0)alpha subunits, we examined the relationship between A beta concentration, G(0)alpha subunit concentration, GTP concentration and rate of GTP hydrolysis experimentally. We found that at low concentrations of A beta (less than 10 microM), A beta increased the rate of GTP hydrolysis over the rate of hydrolysis in the absence of peptide, however, at high concentrations of A beta, significantly decreased rates of GTP hydrolysis were observed. We postulated several molecular level mechanisms for the observed rate behavior, from those mechanisms derived rate equations, and then tested the mechanisms against our experimental rate data. Based on our results, we identified a plausible mechanism for A beta-induced G protein activation which is consistent with available experimental data. This work demonstrates the utility of an engineering approach to examining steps in the mechanism of A beta-induced cell toxicity and could provide insight into our understanding of the mechanism of Alzheimer's disease. PMID- 12628234 TI - A systems interpretation for observations of bird V-formations. AB - Birds in V formations are frequently observed and two main hypotheses have emerged to explain this particular geometry: (i) it offers aerodynamic advantages and (ii) it is used to improve visual communication. Both explanations require a bird to track its predecessor. However, most V-formations observed in nature are small and the distribution of wing-tip spacings has a large variation. This suggests that tracking the lateral position of the preceding bird is a difficult task. Control theorists, when trying to control platoons of vehicles, also noted that predecessor following is difficult. In this paper, we apply a result from systems theory to explain the observations of bird V-formations. The strength of this result is that it does not rely on the details of the bird flight model. Thus we claim that formation flight is inherently difficult for birds. PMID- 12628235 TI - Difference in the retinal cone mosaic pattern between zebrafish and medaka: cell rearrangement model. AB - In fish retina, four kinds of photoreceptor cells (or cones) are two dimensionally arranged in a very regular manner, forming cone mosaics. Mosaic pattern differs between species--two typical patterns are "row mosaic" and "square mosaic", exemplified by the cone mosaics in zebrafish and in medaka, respectively. In this paper, we study a cell-rearrangement model. Cells with pre fixed fate exchange their locations between nearest neighbors and form regular mosaic patterns spontaneously, if the adhesive force between nearest neighbors and between next-nearest neighbors depend on their cell types in an appropriate manner. The same model can produce both row and square mosaic patterns. However, if the cell-cell interaction is restricted to nearest neighbors only, the square mosaic (medaka pattern) cannot be generated, showing the importance of interaction between next-nearest neighbors. In determining whether row mosaic (zebrafish pattern) or square mosaic (medaka pattern) is to be formed, two shape factors are very important, which control the way adhesions in different geometric relations are combined. We also developed theoretical analysis of the parameter ranges for the row mosaic and the square mosaic to have higher total adhesion than alternative spatial patterns. PMID- 12628236 TI - A note on metabolic rate dependence on body size in plants and animals. PMID- 12628237 TI - Sequence context at human single nucleotide polymorphisms: overrepresentation of CpG dinucleotide at polymorphic sites and suppression of variation in CpG islands. AB - Human polymorphisms originate as mutations, and the influence of context on mutagenesis should be reflected in the distribution of sequences surrounding single nucleotide polymorphisms (SNPs). We have performed a computational survey of nearly two million human SNPs to determine if sequence-dependent hotspots for polymorphism exist in the human genome. Here we show that sequences containing CpG dinucleotides, which occur at low frequencies in the human genome, are 6.7 fold more abundant at polymorphic sites than expected. In contrast, polymorphisms in CpG sequences located within CpG islands, important regulatory regions that modulate gene expression, are 6.8-fold less prevalent than expected. The distribution of polymorphic alleles at CpGs in CpG islands is also significantly different from that in non-island regions. These data strongly support a role for 5-methylcytosine deamination in the generation of human variation, and suggest that variation at CpGs in islands is suppressed. PMID- 12628238 TI - Locking TolC entrance helices to prevent protein translocation by the bacterial type I export apparatus. AB - The periplasmic entrance of the TolC channel tunnel is sealed by close-packing of inner and outer coiled-coils, and it has been proposed that opening of the entrance is achieved by an iris-like realignment of the inner coiled-coils. This is supported by experimental disruption of the key links connecting them, which effects transition to the open state in TolC inserted into planar lipid bilayers. Here we provide in vivo evidence for this "twist to open" mechanism by constraining the coiled coils with disulphide bonds, either self-locking or bridged by a chemical cross-linker, and reconstituting the resulting TolC variants into the type I protein export system in Escherichia coli. Introducing an intermonomer disulphide bridge between Ala159 and Ser350 caused a fivefold reduction in export, and when the coiled coils were cross-linked at the entrance constriction, between Asp374 of adjacent monomers or between Asn156 and Ala375, TolC-dependent export was abolished. In vivo cross-linking showed that the locked non-exporting TolC variants were still recruited to assemble the type I export apparatus. The data show that untwisting the entrance helices is essential for the export function of TolC in E.coli, specifically to allow access and passage of substrates engaged at the inner membrane translocase. PMID- 12628239 TI - The crystal structure of R-specific alcohol dehydrogenase from Lactobacillus brevis suggests the structural basis of its metal dependency. AB - The crystal structure of the apo-form of an R-specific alcohol dehydrogenase from Lactobacillus brevis (LB-RADH) was solved and refined to 1.8A resolution. LB-RADH is a member of the short-chain dehydrogenase/reductase (SDR) enyzme superfamily. It is a homotetramer with 251 amino acid residues per subunit and uses NADP(H) as co-enzyme. NADPH and the substrate acetophenone were modelled into the active site. The enantiospecificity of the enzyme can be explained on the basis of the resulting hypothetical ternary complex. In contrast to most other SDR enzymes, the catalytic activity of LB-RADH depends strongly on the binding of Mg(2+). Mg(2+) removal by EDTA inactivates the enzyme completely. In the crystal structure, the Mg(2+)-binding site is well defined. The ion has a perfect octahedral coordination sphere and occupies a special position concerning crystallographic and molecular point symmetry, meaning that each RADH tetramer contains two magnesium ions. The magnesium ion is no direct catalytic cofactor. However, it is structurally coupled to the putative C-terminal hinge of the substrate-binding loop and, via an extended hydrogen bonding network, to some side-chains forming the substrate binding region. Therefore, the presented structure of apo-RADH provides plausible explanations for the metal dependence of the enzyme. PMID- 12628240 TI - Unmasking ligand binding motifs: identification of a chemokine receptor motif by NMR studies of antagonist peptides. AB - Determining the critical structural features a ligand must possess in order to bind to its receptor is of key importance to the understanding of vital biological processes and to the rational design of small molecule therapeutics to modulate receptor function. We have developed a general strategy for determining such ligand binding motifs using low temperature NMR structures of peptides with the desired receptor binding properties. This approach has been successfully applied to determine a binding motif for the chemokine receptor CXCR4. The motif identified provides a detailed guide for the design of small molecule antagonists against CXCR4, which are much sought after to aid in the treatment of a number of conditions including human immunodeficiency virus type 1 infection and a variety of cancers. PMID- 12628241 TI - Twelve new MotA-dependent middle promoters of bacteriophage T4: consensus sequence revised. AB - Bacteriophage T4 middle-mode transcription requires Escherichia coli RNA polymerase, phage-encoded transcriptional activator MotA and co-activator AsiA that form a complex at a middle promoter DNA. T4 middle promoters have been defined by a consensus sequence deduced from the list of 14 middle promoters identified in earlier studies. To date, 33 middle promoters have been mapped on the T4 genome. Of these, 12 contain differences even at the highly conserved positions of the consensus sequence. In the T4 prereplicative gene cluster between genes e and rpbA, we have identified 12 new middle promoters, most of which contain differences from the consensus sequence deduced previously. Analysis of base conservation in the different sequence positions of new middle promoters, as well as those identified previously, revealed some new features of middle T4 promoters. We propose to define these promoters by a MotA box (a/t)(a/t)(a/t)TGCTTtA centred at the position -30, the sequence TAtaAT centred at -10 relative to the transcriptional start site, and the spacer region of 12(+/ 1) base-pairs between them. PMID- 12628242 TI - Unique amino acid composition of proteins in halophilic bacteria. AB - The amino acid compositions of proteins from halophilic archaea were compared with those from non-halophilic mesophiles and thermophiles, in terms of the protein surface and interior, on a genome-wide scale. As we previously reported for proteins from thermophiles, a biased amino acid composition also exists in halophiles, in which an abundance of acidic residues was found on the protein surface as compared to the interior. This general feature did not seem to depend on the individual protein structures, but was applicable to all proteins encoded within the entire genome. Unique protein surface compositions are common in both halophiles and thermophiles. Statistical tests have shown that significant surface compositional differences exist among halophiles, non-halophiles, and thermophiles, while the interior composition within each of the three types of organisms does not significantly differ. Although thermophilic proteins have an almost equal abundance of both acidic and basic residues, a large excess of acidic residues in halophilic proteins seems to be compensated by fewer basic residues. Aspartic acid, lysine, asparagine, alanine, and threonine significantly contributed to the compositional differences of halophiles from meso- and thermophiles. Among them, however, only aspartic acid deviated largely from the expected amount estimated from the dinucleotide composition of the genomic DNA sequence of the halophile, which has an extremely high G+C content (68%). Thus, the other residues with large deviations (Lys, Ala, etc.) from their non halophilic frequencies could have arisen merely as "dragging effects" caused by the compositional shift of the DNA, which would have changed to increase principally the fraction of aspartic acid alone. PMID- 12628243 TI - Differences between the interaction of beta-catenin with non-phosphorylated and single-mimicked phosphorylated 20-amino acid residue repeats of the APC protein. AB - The tumour suppressor protein adenomatous polyposis coli (APC) regulates the level and the intracellular localisation of the proto-oncoprotein beta-catenin. There are indications that a region comprising seven homologous 20-amino acid residue repeats within the APC protein is responsible for the interaction with beta-catenin and that the phosphorylation of conserved serine residues within these repeats increases the affinity for beta-catenin. We used biophysical methods to analyse the beta-catenin binding of single repeats or repeat combinations as non-phosphorylated or phosphorylated recombinant proteins. The non-phosphorylated repeats showed similar affinities, no matter whether they were tested as single recombinant repeats or in combination with neighbouring repeats. This result makes a cooperative influence between the repetitive motifs unlikely. The phosphorylation of the APC protein was mimicked by specific serine/aspartate mutations, which align to serine residues in the cytoplasmic beta-catenin binding domain of E-cadherin. Remarkably, the mimicked phosphorylation of a serine, which is not involved in beta-catenin interaction in the E-cadherin/beta-catenin complex, led to a significant increase in the APC affinity for beta-catenin. These results indicate structural differences between the E-cadherin/beta-catenin and the APC/beta-catenin complexes and provide quantitative evidence for the importance of the APC phosphorylation for its interaction with beta-catenin. PMID- 12628244 TI - Rapid kinetic analysis of EF-G-dependent mRNA translocation in the ribosome. AB - Precise and coordinated movement of the tRNA-mRNA complex within the ribosome is a fundamental step during protein biosynthesis. The molecular mechanism for this process is still poorly understood. Here we describe a new sensitive method for monitoring elongation factor G-dependent translocation of the mRNA in the ribosome. In this method, the fluorescent probe pyrene is covalently attached to the 3' end of a short mRNA sequence at position +9. Translocation of the mRNA by one codon results in a significant decrease in the fluorescence emission of pyrene and can be used to directly monitor mRNA movement using rapid kinetic methods. Importantly, this method offers the flexibility of using any tRNA or tRNA analog in order to elucidate the molecular mechanism of translocation. Our results show that the mRNA is translocated at the same rate as the tRNAs, which is consistent with the view that the movement of the tRNAs and the mRNA are coupled in the ribosome. Furthermore, an anticodon stem-loop analog of tRNA is translocated from the ribosomal A site at a rate constant that is 350-fold lower than peptidyl tRNA, indicating that the D stem, T stem and acceptor stem of A site tRNA contribute significantly to the rate of translocation. PMID- 12628245 TI - Engineering of restriction endonucleases: using methylation activity of the bifunctional endonuclease Eco57I to select the mutant with a novel sequence specificity. AB - Type II restriction endonucleases (REs) are widely used tools in molecular biology, biotechnology and diagnostics. Efforts to generate new specificities by structure-guided design and random mutagenesis have been unsuccessful so far. We have developed a new procedure called the methylation activity-based selection (MABS) for generating REs with a new specificity. MABS uses a unique property of bifunctional type II REs to methylate DNA targets they recognize. The procedure includes three steps: (1) conversion of a bifunctional RE into a monofunctional DNA-modifying enzyme by cleavage center disruption; (2) mutagenesis and selection of mutants with altered DNA modification specificity based on their ability to protect predetermined DNA targets; (3) reconstitution of the cleavage center's wild-type structure. The efficiency of the MABS technique was demonstrated by altering the sequence specificity of the bifunctional RE Eco57I from 5'-CTGAAG to 5'-CTGRAG, and thus generating the mutant restriction endonuclease (and DNA methyltransferase) of a specificity not known before. This study provides evidence that MABS is a promising technique for generation of REs with new specificities. PMID- 12628246 TI - The energetics of specific binding of AT-hooks from HMGA1 to target DNA. AB - The interaction of the second and third AT-hooks of HMGA1 (formerly HMGI/Y), which bind selectively in the minor groove of an AT-rich DNA sequence, was studied at different temperatures and ionic strengths by spectropolarimetry, spectrofluorimetry, isothermal titration calorimetry and differential scanning calorimetry. The data show that binding of the ten amino acid core element of the two AT-hooks, which penetrates deep into the minor groove, is entropically driven: both the entropy and enthalpy of association of the peptides to the target DNA are positive up to 50 degrees C. The seven amino acid extension of the core in the second AT-hook, which extends out from the minor groove and loops over the phosphodiester backbone, adds a substantial negative enthalpic component into the binding of the 17 residue DBD2 peptide to DNA that corresponds in magnitude to the enthalpy of formation of two hydrogen bonds. The ionic strength dependence of the association constant allowed an estimation of the electrostatic component of binding and, by subtraction, the contribution of the non electrostatic component, which results from dehydration of the contacting surfaces and makes up almost 70% of the total energy of complex formation. The exceptionally large positive entropy and enthalpy of association of the core AT hook peptides with target DNA suggest that the water, which is removed from the minor groove of DNA upon binding, is in a highly ordered state. Acetylation of the lysine residue in the second AT-hook, which corresponds to Lys65 of HMGA1, has little effect on the DNA binding; so it appears that repression of the hIFNbeta gene, which follows this modification, is not a direct result of the abrogation of DNA binding. PMID- 12628247 TI - Mechanism of inhibition of site-specific recombination by the Holliday junction trapping peptide WKHYNY: insights into phage lambda integrase-mediated strand exchange. AB - Holliday junctions are central intermediates in site-specific recombination reactions mediated by tyrosine recombinases. Because these intermediates are extremely transient, only artificially assembled Holliday junctions have been available for study. We have recently identified hexapeptides that cause the accumulation of natural Holliday junctions of bacteriophage lambda Integrase (Int)-mediated reactions. We now show that one of these peptides acts after the first DNA cleavage event to stabilize protein-bound junctions and to prevent their resolution. The peptide acts before the step affected by site affinity (saf) mutations in the core region, in agreement with a model that the peptide stabilizes the products of strand exchange (i.e. Holliday junctions) while saf mutations reduce ligation of exchanged strands.Strand exchange events leading to Holliday junctions in phage lambda integration and excision are asymmetric, presumably because interactions between Int and some of its core-binding sites determine the order of strand cleavage. We have compared the structure of Holliday junctions in one unidirectional and in two bidirectional Int-mediated pathways and show that the strand cleavage steps are much more symmetric in the bidirectional pathways. Thus Int-DNA interactions which determine the order of top and bottom strand cleavage and exchange are unique in each recombination pathway. PMID- 12628248 TI - Insight into the functional consequences of inherited variants of the hMYH adenine glycosylase associated with colorectal cancer: complementation assays with hMYH variants and pre-steady-state kinetics of the corresponding mutated E.coli enzymes. AB - The oxidized guanine lesion 7,8-dihydro-8-oxo-2'-deoxyguanosine (OG) is highly mutagenic, resulting in G:C to T:A transversion mutations in the absence of repair. The Escherichia coli adenine glycosylase MutY and its human homolog (hMYH) play an important role in the prevention of mutations associated with OG by removing misincorporated adenine residues from OG:A mismatches. Previously, biallelic mutations of hMYH have been identified in a British family (Family N) with symptoms characteristic of familial adenomatous polyposis (FAP), which is typically associated with mutations in the adenomatous polyposis coli (APC) gene. Afflicted members of this family were compound heterozygotes for two mutations in hMYH, Y165C and G382D. These positions are highly conserved in MutY across phylogeny. The current work reveals a reduced ability of the hMYH variants compared to wild-type (WT) hMYH to complement the activity of E.coli MutY in mutY((-)) E.coli. In vitro analysis of the corresponding mutations in E.coli MutY revealed a reduction in the adenine glycosylase activity of the enzymes. In addition, evaluation of substrate affinity using a substrate analog, 2'-deoxy-2' fluoroadenosine (FA) revealed that both mutations severely diminish the ability to recognize FA, and discriminate between OG and G. Importantly, adenine removal with both the mutant and WT E.coli enzymes was observed to be less efficient from a mismatch in the sequence context observed to be predominantly mutated in tumors of Family N. Interestingly, the magnitude of the reduced activity of the E.coli mutant enzymes relative to the WT enzyme was magnified in the "hotspot" sequence context. If the corresponding mutations in hMYH cause similar sensitivity to sequence context, this effect may contribute to the specific targeting of the APC gene. The lack of complementation of the hMYH variants for MutY, and the reduced activity of the Y82C and G253D E.coli enzymes, provide additional circumstantial evidence that the somatic mutations in APC, and the occurrence of FAP in Family N, are due to a reduced ability of the Y165C and G382D hMYH enzymes to recognize and repair OG:A mismatches. PMID- 12628249 TI - Design of a modular immunotoxin connected by polyionic adapter peptides. AB - Immunotoxins are genetically engineered fusion proteins of an antibody Fv fragment and a toxin from bacteria or plants, which function as anti-cancer therapeutics. Here, we describe a new generation of immunotoxins in which both proteins do not form a single fusion protein but are coupled specifically via cysteine-containing polyionic fusion peptides. The engineered Pseudomonas exotoxin PE38 was N-terminally fused to the peptide E(8)C. In combination with the disulfide-stabilized Fv fragment of the tumor-specific antibody B3, which was extended by the peptide R(8)CP, the fusion peptides ensured a specific and covalent coupling of the Fv fragment and the toxin. The resulting immunotoxin was as active and as specific as an immunotoxin consisting of a fusion protein of the same antibody fragment connected to the toxin. PMID- 12628251 TI - Biogenesis of yeast mitochondrial cytochrome c: a unique relationship to the TOM machinery. AB - The import of cytochrome c into the mitochondrial intermembrane space is not understood at a mechanistic level. While the precursor apocytochrome c can insert into protein-free lipid bilayers, the purified translocase of the outer membrane (TOM) complex supports the translocation of apocytochrome c into proteoliposomes. We report an in organello analysis of cytochrome c import into yeast mitochondria from wild-type cells and different mutants cells, each defective in one of the seven Tom proteins. The import of cytochrome c is not affected by removal of the receptor Tom20 or Tom70. Moreover, neither the transfer protein Tom5 nor the assembly factors Tom6 and Tom7 are needed for import of cytochrome c. When the general import pore (GIP)-protein Tom40 is blocked, the import of cytochrome c is moderately affected. Mitochondria lacking the central receptor and organizing protein Tom22 contain greatly reduced levels of cytochrome c. We conclude that up to two components of the TOM complex, Tom22 and possibly the GIP, are involved in the biogenesis of cytochrome c. PMID- 12628250 TI - Ion-coupling determinants of Na+-driven and H+-driven flagellar motors. AB - The bacterial flagellar motor is a tiny molecular machine that uses a transmembrane flux of H(+) or Na(+) ions to drive flagellar rotation. In proton driven motors, the membrane proteins MotA and MotB interact via their transmembrane regions to form a proton channel. The sodium-driven motors that power the polar flagellum of Vibrio species contain homologs of MotA and MotB, called PomA and PomB. They require the unique proteins MotX and MotY. In this study, we investigated how ion selectivity is determined in proton and sodium motors. We found that Escherichia coli MotA/B restore motility in DeltapomAB Vibrio alginolyticus. Most hypermotile segregants isolated from this weakly motile strain contain mutations in motB. We constructed proteins in which segments of MotB were fused to complementary portions of PomB. A chimera joining the N terminus of PomB to the periplasmic C terminus of MotB (PotB7(E)) functioned with PomA as the stator of a sodium motor, with or without MotX/Y. This stator (PomA/PotB7(E)) supported sodium-driven motility in motA or motB E.coli cells, and the swimming speed was even higher than with the original stator of E.coli MotA/B. We conclude that the cytoplasmic and transmembrane domains of PomA/B are sufficient for sodium-driven motility. However, MotA expressed with a B subunit containing the N terminus of MotB fused to the periplasmic domain of PomB (MomB7(E)) supported sodium-driven motility in a MotX/Y-dependent fashion. Thus, although the periplasmic domain of PomB is not necessary for sodium-driven motility in a PomA/B motor, it can convert a MotA/B proton motor into a sodium motor. PMID- 12628252 TI - Catalase-peroxidase KatG of Burkholderia pseudomallei at 1.7A resolution. AB - The catalase-peroxidase encoded by katG of Burkholderia pseudomallei (BpKatG) is 65% identical with KatG of Mycobacterium tuberculosis, the enzyme responsible for the activation of isoniazid as an antibiotic. The structure of a complex of BpKatG with an unidentified ligand, has been solved and refined at 1.7A resolution using X-ray synchrotron data collected from crystals flash-cooled with liquid nitrogen. The crystallographic agreement factors R and R(free) are 15.3% and 18.6%, respectively. The crystallized enzyme is a dimer with one modified heme group and one metal ion, likely sodium, per subunit. The modification on the heme group involves the covalent addition of two or three atoms, likely a perhydroxy group, to the secondary carbon atom of the vinyl group on ring I. The added group can form hydrogen bonds with two water molecules that are also in contact with the active-site residues Trp111 and His112, suggesting that the modification may have a catalytic role. The heme modification is in close proximity to an unusual covalent adduct among the side-chains of Trp111, Tyr238 and Met264. In addition, Trp111 appears to be oxidized on C(delta1) of the indole ring. The main channel, providing access of substrate hydrogen peroxide to the heme, contains a region of unassigned electron density consistent with the binding of a pyridine nucleotide-like molecule. An interior cavity, containing the sodium ion and an additional region of unassigned density, is evident adjacent to the adduct and is accessible to the outside through a second funnel shaped channel. A large cleft in the side of the subunit is evident and may be a potential substrate-binding site with a clear pathway for electron transfer to the active-site heme group through the adduct. PMID- 12628253 TI - The crystal structure of aminoglycoside-3'-phosphotransferase-IIa, an enzyme responsible for antibiotic resistance. AB - A major factor in the emergence of antibiotic resistance is the existence of enzymes that chemically modify common antibiotics. The genes for these enzymes are commonly carried on mobile genetic elements, facilitating their spread. One such class of enzymes is the aminoglycoside phosphotransferase (APH) family, which uses ATP-mediated phosphate transfer to chemically modify and inactivate aminoglycoside antibiotics such as streptomycin and kanamycin. As part of a program to define the molecular basis for aminoglycoside recognition and inactivation by such enzymes, we have determined the high resolution (2.1A) crystal structure of aminoglycoside-3'-phosphotransferase-IIa (APH(3')-IIa) in complex with kanamycin. The structure was solved by molecular replacement using multiple models derived from the related aminoglycoside-3'-phosphotransferase-III enzyme (APH(3')-III), and refined to an R factor of 0.206 (R(free) 0.238). The bound kanamycin molecule is very well defined and occupies a highly negatively charged cleft formed by the C-terminal domain of the enzyme. Adjacent to this is the binding site for ATP, which can be modeled on the basis of nucleotide complexes of APH(3')-III; only one change is apparent with a loop, residues 28 34, in a position where it could fold over an incoming nucleotide. The three rings of the kanamycin occupy distinct sub-pockets in which a highly acidic loop, residues 151-166, and the C-terminal residues 260-264 play important parts in recognition. The A ring, the site of phosphoryl transfer, is adjacent to the catalytic base Asp190. These results give new information on the basis of aminoglycoside recognition, and on the relationship between this phosphotransferase family and the protein kinases. PMID- 12628254 TI - High-resolution X-ray and NMR structures of the SMN Tudor domain: conformational variation in the binding site for symmetrically dimethylated arginine residues. AB - The SMN protein, which is linked to spinal muscular atrophy (SMA), plays an important role in the assembly of the spliceosomal small nuclear ribonucleoprotein complexes. This function requires binding of SMN to the arginine-glycine (RG) rich C-terminal tails of the Sm proteins, which contain symmetrically dimethylated arginine residues (sDMA) in vivo. Using NMR titrations, we show that the SMN Tudor domain recognizes these sDMAs in the methylated RG repeats. Upon complex formation a cluster of conserved aromatic residues in the SMN Tudor domain interacts with the sDMA methyl groups. We present two high resolution structures of the uncomplexed SMN Tudor domain, a 1.8A crystal structure and an NMR structure that has been refined against a large number of backbone and side-chain residual dipolar couplings. The backbone conformation of both structures is very similar, however, differences are observed for the cluster of conserved aromatic side-chains in the sDMA binding pocket. In order to validate these variations we introduce a novel application of residual dipolar couplings for aromatic rings. We show that structural information can be derived from aromatic ring residual dipolar couplings, even in the presence of internal motions such as ring flipping. These residual dipolar couplings and ring current shifts independently confirm that the SMN Tudor domain adopts two different conformations in the sDMA binding pocket. The observed structural variations may play a role for the recognition of sDMAs. PMID- 12628255 TI - Solution NMR structure of ribosome-binding factor A (RbfA), a cold-shock adaptation protein from Escherichia coli. AB - Ribosome-binding factor A (RbfA) from Escherichia coli is a cold-shock adaptation protein. It is essential for efficient processing of 16S rRNA and is suspected to interact with the 5'-terminal helix (helix I) of 16S rRNA. RbfA is a member of a large family of small proteins found in most bacterial organisms, making it an important target for structural proteomics. Here, we describe the three dimensional structure of RbfADelta25, a 108 residue construct with 25 residues removed from the carboxyl terminus of full-length RbfA, determined in solution at pH 5.0 by heteronuclear NMR methods. The structure determination was carried out using largely automated methods for determining resonance assignments, interpreting nuclear Overhauser effect (NOE) spectroscopy (NOESY) spectra, and structure generation. RbfADelta25 has an alpha+beta fold containing three helices and three beta-strands, alpha1-beta1-beta2-alpha2-alpha3-beta3. The structure has type-II KH-domain fold topology, related to conserved KH sequence family proteins whose betaalphaalphabeta subunits are characterized by a helix-turn-helix motif with sequence signature GxxG at the turn. In RbfA, this betaalphaalphabeta subunit is characterized by a helix-kink-helix motif in which the GxxG sequence is replaced by a conserved AxG sequence, including a strongly conserved Ala residue at position 75 forming an interhelical kink. The electrostatic field distribution about RbfADelta25 is bipolar; one side of the molecule is strongly negative and the opposite face has a strong positive electrostatic field. A "dynamic hot spot" of RbfADelta25 has been identified in the vicinity of a beta bulge at strongly conserved residue Ser39 by 15N R(1), R(2) relaxation rate and heteronuclear 15N-1H NOE measurements. Analyses of these distributions of electrostatic field and internal dynamics, together with evolutionary implications of fold and sequence conservation, suggest that RbfA is indeed a nucleic acid-binding protein, and identify a potential RNA-binding site in or around the conserved polypeptide segment Ser76-Asp100 corresponding to the alpha3 loop-beta3 helix-loop-strand structure. While the structure of RbfADelta25 is most similar to that of the KH domain of the E.coli Era GTPase, its electrostatic field distribution is most similar to the KH1 domain of the NusA protein from Thermotoga maritima, another cold-shock associated RNA-binding protein. Both RbfA and NusA are regulated in the same E.coli operon. Structural and functional similarities between RbfA, NusA, and other bacterial type II KH domains suggest previously unsuspected evolutionary relationships between these cold-shock associated proteins. PMID- 12628257 TI - Importance of substrate and cofactor polarization in the active site of dihydrofolate reductase. AB - By using a combined quantum-mechanical and molecular-mechanical potential in molecular dynamics simulations, we have investigated the effects of the enzyme electric field of dihydrofolate reductase on the electronic polarization of its 5 protonated dihydrofolate substrate at various stages of the catalyzed hydride transfer reaction. Energy decomposition of the total electrostatic interaction energy between the ligands and the enzyme shows that the polarization effect is 4% of the total electrostatic interaction energy, and, significantly, it accounts for 9kcal/mol of transition state stabilization relative to the reactant state. Therefore it is essential to take account of substrate polarization for quantitative interpretation of enzymatic function and for calculation of binding free energies of inhibitors to a protein. Atomic polarizations are calculated as the differences in the average atomic charges on the atoms in gas phase and in molecular simulations of the enzyme; this analysis shows that the glutamate tail and the pterin ring are the highly polarized regions of the substrate. Electron density difference plots of the reactant and product complexes at instantaneous configurations in the enzyme active center confirm the inferences made on the basis of partial atomic charges. PMID- 12628256 TI - Thermodynamics of a designed protein catenane. AB - Topological linking of proteins is a new approach for stabilizing and controlling the oligomerization state of proteins that fold in an interwined manner. The recent design of a backbone cyclized protein catenane based on the p53tet domain suggested that topological cross-linking provided increased stability against thermal and chemical denaturation. However, the tetrameric structure complicated detailed biophysical analysis of this protein. Here, we describe the design, synthesis and thermodynamic characterization of a protein catenane based on a dimeric mutant of the p53tet domain (M340E/L344K). The formation of the catenane proceeded efficiently, and the overall structure and oligomerization of the domain was not affected by the formation of the topological link. Unfolding and refolding of the catenane was consistent with a two-state process. The topological link stabilized the dimer against thermal and chemical denaturation considerably, raising the apparent melting temperature by 59 degrees C and the midpoint of denaturation by 4.5M GuHCl at a concentration of 50 microM. The formation of the topological link increased the resistance of the dimer to proteolysis. However, the m value decreased by 1.7kcalmol(-1)M(-1), suggesting a decrease in accessible surface area in the unfolded state. This implies that the stabilization from the topological link is largely due to a destabilization of the unfolded state, similar to other cross-links in proteins. Topological linking therefore provides a powerful and orthogonal tool for the stabilization of peptide and protein oligomers. PMID- 12628259 TI - Laparoscopic colposuspension: a systematic review. AB - OBJECTIVE: To determine the effectiveness of laparoscopic colposuspension for the treatment of stress urinary incontinence. DESIGN: Systematic review. SETTING: Teaching hospitals. POPULATION: Women with symptoms or urodynamic diagnosis of stress, urge or mixed incontinence. METHODS: Randomised and 'quasi-randomised trials' in women with symptoms or urodynamic diagnoses of stress, urge or mixed incontinence including laparoscopic surgery in at least one arm of the trial were identified from the Cochrane Incontinence Review Group's Specialised Register of Controlled Trials. MAIN OUTCOME MEASURES: Subjective cure, objective cure, quality of life measurements and surgical outcome measures. RESULTS: Eight trials met the inclusion criteria. Five trials compared laparoscopic (n = 233) with open (n = 254) colposuspension. Risk of a positive stress test at follow up was significantly less in the open group (relative risk [RR] 0.89, 95% confidence interval [CI] 0.82 to 0.98). When one poor quality trial was excluded from the analysis, the relative risk was still less in the open group, but was no longer significant (RR 0.91, 95% CI 0.82 to 1.01). If considered as risk difference there is a 9% significantly higher risk of failure for laparoscopic colposuspension, however, excluding one poor quality trial, this risk of failure is non-significant at 8%. Analysis of subjective cure showed no difference between the groups. A trend was shown towards a higher complication rate, less post-operative pain, shorter hospital stay and time to return to normal function for laparoscopic compared with open colposuspension. Three trials compared different methods of laparoscopic colposuspension. Data from these trials could not be combined in view of the differences in the trials. Based on a single trial, two sutures were better than one. Sutures were as effective as mesh/staples. No conclusion could be drawn about the benefits of transperitoneal or extraperitoneal access. CONCLUSIONS: It is difficult to make valid conclusions as the evidence is limited by short term follow up and small numbers of trials and participants. However, it is possible that no difference exists between laparoscopic and open colposuspension for either objective or subjective cure. Further, well designed and adequately powered randomised controlled trials are required. PMID- 12628260 TI - Laparoscopic myomectomy for fibroids penetrating the uterine cavity: is it a safe procedure? AB - OBJECTIVE: The purpose of the study was to evaluate the post-operative course and follow up of women who had undergone laparoscopic removal of intramural fibroids penetrating the uterine cavity. DESIGN: Retrospective study. SETTING: Center for Reconstructive Pelvic Endosurgery, Italy. POPULATION: Thirty-four women with fibroids penetrating the uterine cavity. METHODS: Laparoscopic myomectomy. MAIN OUTCOME MEASURES: Feasibility and safety of surgical technique, length of operation, blood loss, intra- or post-operative complications, length of hospital stay, resolution of symptoms and future obstetric outcome. RESULTS: The mean operative time was 79 (SD 30) minutes; the mean reduction in haemoglobin was 1.1 +/- 0.9 g/dL. No intra- or post-operative complications were observed. The average post-operative stay in hospital was 54 (SD 22) hours. Nineteen (73%) out of 26 patients who had experienced symptoms prior to surgery reported resolution of these symptoms post-operatively. All patients resumed work within a mean time of 20 (SD 8) days. Among 23 of the 32 patients attempting pregnancy during the follow up period, nine (39%) conceived within one year. Seven pregnancies went to term without complications. CONCLUSIONS: The clinical results of this study suggest that laparoscopic myomectomy for intramural fibroids penetrating the uterine cavity is a safe procedure, providing well known advantages of minimal access surgery. PMID- 12628261 TI - Early invasive cervical adenocarcinoma: its potential for nodal metastasis or recurrence. AB - OBJECTIVE: To investigate the potential for nodal spread or recurrence in patients with early invasive cervical adenocarcinoma. The possible application of the International Federation of Gynecology and Obstetrics (FIGO) classification (1994) to this variant was also examined. DESIGN: Retrospective observational study. SETTING: Gynaecological oncology division of Cancer Institute Hospital, Japan. POPULATION: 302 patients with FIGO Stage 0-IIB cervical adenocarcinoma treated surgically at the Cancer Institute Hospital. METHODS: Clinicopathological analysis was performed on 47 patients with early invasive cervical adenocarcinoma in whom the depth of stromal invasion was 5 mm or less. All patients underwent radical hysterectomy and pelvic lymphadenectomy. RESULTS: In 30 patients with a depth of tumour invasion of 3 mm or less, no lymph node metastasis was found, while two patients developed recurrence; one had a depth of invasion of 3 mm and a horizontal tumour spread of 3 mm, and the other had horizontal spread of more than 7 mm. In 17 patients with a depth of invasion from 3 to 5 mm, there was also no lymph node metastasis, but two patients developed recurrence; one had horizontal tumour spread of 7 mm or less, and the other had horizontal spread of more than 7 mm. CONCLUSIONS: Early invasive cervical adenocarcinoma with a depth of invasion of 3 mm or less and a horizontal spread of 7 mm or less has little potential for nodal metastasis or recurrence. It seems possible that the FIGO definition (1994) of early cervical cancer may be applicable in its present form to early cervical adenocarcinoma. PMID- 12628262 TI - Are smoking and other lifestyle factors associated with female urinary incontinence? The Norwegian EPINCONT Study. AB - OBJECTIVE: To examine whether modifiable lifestyle factors such as smoking, obesity, physical activity and intake of alcohol or caffeinated drinks were associated with urinary incontinence in women. DESIGN: Cross sectional population based study. SETTING: The Norwegian Epidemiology of Incontinence in the County of Nord-Trondelag (EPINCONT) Study is part of a large survey performed in a county in Norway during 1995-1997. POPULATION: Women >/=20 years (n = 34,755, 75% of the invited) attended the first part of the survey and received the questionnaire. There were 27,936 (80% of source population) women who completed the incontinence part of the questionnaire. METHODS: Questionnaire covering several health topics including urinary incontinence was received at a screening station. Logistic regression analysis was used to adjust for confounding and to establish associations with the different outcomes under investigation: any incontinence, severe incontinence and stress, urge and mixed subtypes. MAIN OUTCOME MEASURES: Effect measure were odds ratios with corresponding 95% confidence intervals. RESULTS: Former and current smoking was associated with incontinence, but only for those who smoked more than 20 cigarettes per day. Severe incontinence was weakly associated with smoking regardless of number of cigarettes. The association between increasing body mass index and incontinence was strong and present for all subtypes. Increasing levels of low intensity physical activity had a weak and negative association with incontinence. Tea drinkers were at slightly higher risk for all types of incontinence. We found no important effects of high intensity physical activity, intake of alcohol or coffee. CONCLUSIONS: Several potentially modifiable lifestyle factors are associated with urinary incontinence. Highest odds ratios were found for body mass index, heavy smoking and tea drinking. PMID- 12628263 TI - A cost-utility analysis of tension-free vaginal tape versus colposuspension for primary urodynamic stress incontinence. AB - OBJECTIVE: To assess the cost effectiveness of tension-free vaginal tape compared with open Burch colposuspension as a primary treatment for urodynamic stress incontinence. DESIGN: Cost-utility analysis alongside a multicentre randomised comparative trial. SETTING: Gynaecology or Urology departments in 14 centres in the UK and Ireland, including University-associated teaching hospitals and district general hospitals. POPULATION: Women with urodynamic stress incontinence. Exclusion criteria were: (1) detrusor overactivity; (2) major voiding problems; (3) prolapse; (4) previous surgery for incontinence or prolapse. METHODS: Resource use data were collected on all 344 patients in the trial, including length of hospital stay, time in theatre and management of complications; resource use was costed using UK unit costs at 1999-2000 prices. MAIN OUTCOME MEASURES: Health outcomes were expressed in terms of quality adjusted life years (QALYs) between baseline and six months follow up, based on women's responses to the EQ-5D health questionnaire. RESULTS: Tension-free vaginal tape resulted in a mean cost saving of pound 243 (95% CI pound 341 to pound 201) compared with colposuspension. Differential mean QALYs per patient (tension-free vaginal tape - colposuspension) was 0.01 (95% CI -0.01 to 0.03). The probability of tension-free vaginal tape being, on average, less costly than colposuspension, was 100%, and the probability of tension-free vaginal tape being more cost effective than colposuspension was 94.6% if the decision-maker was willing to pay pound 30,000 per additional QALY. CONCLUSIONS: The results from this trial suggest that, over a post-operative period of six months, tension-free vaginal tape is a cost effective alternative to colposuspension. The results will need to be reassessed on the basis of longer follow up. PMID- 12628264 TI - Choices about abortion method: assessing the quality of patient information leaflets in England and Wales. AB - OBJECTIVE: To assess the quality of patient leaflets regarding the choice between medical and surgical abortions. DESIGN: Cross sectional survey. SETTING: NHS and private providers of abortion services in England and Wales. SAMPLE: Leaflets from three private providers, all teaching hospitals (n = 14) and a representative sample of NHS district hospitals (n = 27), accessed by approximately 100,000 women per year. METHODS: On telephoning the above abortion services, the leaflets provided to women choosing to have medical or surgical abortions were requested. Content analysis of leaflets included: application of a coding frame assessing adequacy of information about options, risks and procedures; the Flesch Readability Ease scale. MAIN OUTCOME MEASURES: Provision of leaflet, adequacy of information and ease of readability. RESULTS: Seventy three percent (44/60) of contacted providers offered women a leaflet. Of these 44 leaflets, one mentioned medical and 15 surgical abortion methods only. Adequacy of information provision was low, with on average 10/23 points covered about the surgical method, 12/21 points about the medical method and 7/11 points about aftercare. Readability scores were poor with 24/44 (55%) rated as 'difficult' or 'fairly difficult'. Among NHS providers, scores were slightly higher in the teaching than in the district hospitals. CONCLUSIONS: On average, leaflets provided half the information possible about the risks and procedures of medical and surgical methods. In addition, readability was rated as difficult in over half the leaflets. It seems unlikely that these leaflets enable women to make informed choices between abortion methods and, therefore, do not meet the requirements of current guidelines. PMID- 12628265 TI - The Haven: a pilot referral centre in London for cases of serious sexual assault. AB - OBJECTIVE: Several schemes have been reported to improve treatment of rape and to encourage reporting. The development of a comprehensive forensic and follow up service for complainants of sexual assault is described, and activities of the first year are reviewed. DESIGN: Retrospective review of case records of complainants examined in The Haven. SETTING: Department of Sexual Health in a London teaching hospital. SAMPLE: All case records, 676 complainants, from the first year of cases seen in The Haven. METHODS: Description of setting up a service in partnership between the National Health Service and the Metropolitan Police, called The Haven. Analysis of a standardised proforma used for case records. RESULTS: Mean age of complainants is 26 years (range 11-66); 6% were male. Assailant was categorised as a stranger in 52% of cases; attack involved physical violence in 50% of cases; 24% of victims had genital injuries; 39% had other physical injuries. Immediate care given at time of forensic examination included 30% of women receiving emergency contraception and 5% of clients receiving post-exposure prophylaxis against HIV. Fifty-five percent of clients returned for a sexual health screen and/or counselling. Thirty-one percent received screening for sexually transmitted infections and 12% were diagnosed with one or more infections. CONCLUSIONS: Requirements following sexual assault include forensic examination, first aid, postcoital contraception, prevention and management of sexually transmitted infections and psychosocial support. Provision of these services within a sexual health setting is feasible. PMID- 12628266 TI - The prevalence of domestic violence in pregnant women. AB - OBJECTIVE: To determine the prevalence of domestic violence in a population of pregnant women. DESIGN: Questionnaire survey. SETTING: Antenatal booking clinic in a north of England hospital. POPULATION: Five hundred consecutive women were included. METHODS: Anonymous confidential questionnaire to women who were not accompanied by their partners. MAIN OUTCOME MEASURES: Disclosure of a past history of physical, emotional or sexual abuse. RESULTS: Four hundred and seventy five questionnaires were returned (95% response rate). The prevalence of domestic violence was 17%. Domestic violence was highest in the age group 26-30 years and boyfriends were the main perpetrators. Punching and slapping were the most common pattern of violence, and 10% of women experiencing domestic violence had had forced sexual activity. CONCLUSIONS: The prevalence of domestic violence in a cohort of pregnant women in the north of England was 17%. Consideration should be given for routine screening for domestic violence in pregnancy to institute effective intervention strategies. PMID- 12628267 TI - Screening for trisomy 21 in twins using first trimester ultrasound and maternal serum biochemistry in a one-stop clinic: a review of three years experience. AB - OBJECTIVE: To evaluate the performance of screening for fetal trisomy 21 in the first trimester of twin pregnancies by a combination of maternal serum biochemistry and ultrasonography. DESIGN: Three year retrospective review of screening performance. SETTING: District General Hospital maternity unit. POPULATION: All women booked to receive routine antenatal care at Harold Wood Hospital between 1 June 1998 and 30th September 2001. The population included 13,940 women of all ages presenting with pregnancies between 10 weeks 3 days and 13 weeks 6 days gestation. Of these, 230 had a twin pregnancy. METHODS: Women booked into the clinic were offered screening using a combination of maternal serum free beta-hCG and pregnancy-associated plasma protein-A (PAPP-A) and fetal nuchal translucency thickness. Women at increased risk of carrying a fetus with trisomy 21 or trisomy 13/18 (>/=1 in 300 at sampling) were offered counselling and an invasive diagnostic procedure. Follow up of the outcome of all pregnancies was carried out. For women who on examination were at 14 weeks of gestation or greater, or for women presenting as late bookers beyond 14 weeks, screening was performed in the same time frame using only maternal serum free beta-hCG and alpha-fetoprotein. MAIN OUTCOME MEASURES: The first trimester detection rate for trisomy 21 and all aneuploides, false positive rate, uptake of screening, uptake of invasive testing in women identified at increased risk and fetal loss rates after invasive testing. RESULTS: Overall, 97.4% of the women with twins (224/230) accepted first trimester screening. The rate of detection of trisomy 21 was 75% (3/4). Fetal death at presentation was found in 3.4% of fetuses (16/460). Of women who accepted screening, 4.3% (10/230) presented too late for fetal nuchal translucency measurement and 10.0% of women (23/230) presented too early. A risk for trisomy 21 was calculated for each fetus based on the individual fetal nuchal translucency thickness and the maternal biochemistry. The false positive rate among those eligible for first trimester screening was 9.0% (19/206) of pregnancies and 6.9% of fetuses (28/412). Uptake of invasive testing was 59% (10/17) with chorionic villus sampling in eight cases and amniocentesis in two. No fetal loss occurred within 28 days of chorionic villus sampling and no loss occurred after amniocentesis. One case of trisomy 21 was identified for every three invasive procedures. CONCLUSIONS: First trimester screening for trisomy 21 in twin pregnancies is both theoretically possible and practically achievable using a combination of nuchal translucency thickness and maternal serum biochemistry. However, dilemmas for the mother and health professionals when both nuchal translucency thickness measurements are normal might suggest that greater reliance be placed on the nuchal translucency thickness risk alone when counselling women about invasive testing. PMID- 12628268 TI - Screening for chromosomal abnormalities in the first trimester using ultrasound and maternal serum biochemistry in a one-stop clinic: a review of three years prospective experience. AB - OBJECTIVE: To evaluate the performance of a one-stop multidisciplinary clinic of screening for fetal chromosomal anomalies in the first trimester of pregnancy by a combination of maternal serum biochemistry and ultrasonography. DESIGN: Retrospective review of screening performance. SETTING: District General Hospital maternity unit. POPULATION: All women booked for routine antenatal care at Harold Wood Hospital between 1 June 1998 and 31 May 2001. The population included 12,339 women with singleton pregnancies presenting at 10-14 weeks of gestation. METHODS: Women were offered screening using a combination of maternal serum free beta-hCG and pregnancy associated plasma protein-A (PAPP-A) and fetal nuchal translucency thickness. Those with an estimated risk of >/=1 in 300 of carrying a fetus with trisomy 21 or trisomy 18 or trisomy 13 were offered the option of an invasive diagnostic test. Follow up of the outcome of all pregnancies was carried out. MAIN OUTCOME MEASURES: Uptake of screening and invasive testing, detection rate for fetal chromosomal abnormalities and false positive rate. RESULTS: The uptake of first trimester screening was 97.5% and the uptake of invasive testing in the increased risk group was 77%. The rate of detection of trisomy 21 was 92% (23 of 25), of trisomy 13 or 18 was 100% (all 15) and of all aneuploidies was 96% (49 of 51). The false positive rate was 5.2%. CONCLUSIONS: First trimester screening for trisomy 21 and other aneuploidies can be delivered in an efficient manner in a one-stop multidisciplinary clinic. The detection rates are far better than can be achieved by second trimester serum screening. PMID- 12628270 TI - Massive perivillous fibrinoid causing recurrent placental failure. AB - OBJECTIVE: To establish the incidence, recurrence rate and consequences of massive perivillous fibrinoid. DESIGN: Retrospective analysis of the histology of all placentas with a diagnosis of massive perivillous fibrinoid between 1991 and 1998, together with the maternal case records. SETTING: The histopathology department of the Rotunda Hospital, Dublin, Ireland. POPULATION: A relatively homogeneous group of pregnant women in the northern part of Dublin City, which is the catchment area for the Rotunda Hospital, delivered between 1991 and 1998. METHODS: Retrospective review of archival placental pathology and maternal charts. MAIN OUTCOME MEASURES: The incidence of massive perivillous fibrinoid, perinatal outcome and recurrence rate. RESULTS: The incidence of massive perivillous fibrinoid was 0.028%, with a recurrence rate of approximately 18%. All the infants suffered intrauterine growth restriction; there was a 31% fetal loss rate and a 33% preterm delivery rate. CONCLUSIONS: Massive perivillous fibrinoid is associated with intrauterine death, intrauterine growth restriction and preterm delivery. It has a significant recurrence rate and both the clinical findings of intrauterine growth restriction and the postmortem findings imply a syndrome of chronic placental insufficiency. PMID- 12628269 TI - Maternal and neonatal hair mercury concentrations: the effect of dental amalgam. AB - OBJECTIVE: To evaluate maternal and fetal hair mercury levels in relation to the placement of dental amalgam tooth restorations. DESIGN: Cross sectional study involving women who never had dental amalgam restorations placed, women who had amalgam restorations placed before pregnancy and women who had restorations placed during the index pregnancy. SETTING: North of England Maternity Hospital. SAMPLE: Fifty-three healthy women who delivered healthy babies at term. METHODS: Maternal and fetal hair was collected in a standardised manner in the first few days following delivery. MAIN OUTCOME MEASURES: Maternal and neonatal hair mercury concentrations. RESULTS: When compared with women without restorations, there was a significant increase in the maternal hair mercury concentration in women who had dental amalgam placed outside of the index pregnancy and also in women who had dental amalgam placed during the index pregnancy. The fetal hair mercury concentration was significantly higher in babies when mothers had been exposed to dental amalgam either before pregnancy or during pregnancy compared with unexposed babies. There was no difference in the maternal or fetal hair mercury levels in the groups of patients who had dental amalgam placed before or during pregnancy. CONCLUSIONS: Maternal and fetal hair mercury levels were significantly higher in women who previously had dental amalgam restorations placed. There was no evidence that placement of dental amalgam restorations in pregnant women who had already similar restorations increased the maternal or fetal hair mercury level. PMID- 12628271 TI - The use of lidocaine for fetocide in late termination of pregnancy. AB - OBJECTIVE: To assess the use of lidocaine (1%) to induce permanent fetal cardiac asystole for fetocide in late termination of pregnancy. DESIGN: Prospective observational study. SETTING: One tertiary referral fetal medicine unit in France. SAMPLE: Fifty patients undergoing termination of pregnancy between 20 and 36 weeks of gestation for severe abnormalities or severe maternal conditions. METHODS: Fetocide was performed by umbilical vein puncture under ultrasound guidance with injection of sufentanil (5 microg) followed by 7 to 30 mL of lidocaine (1%). MAIN OUTCOME MEASURES: Percentage of successful procedures to obtain permanent fetal cardiac asystole and maternal side effects. RESULTS: The procedure was successful in 92% of cases (46/50) with complete cessation of heart activity. The mean amount of lidocaine was 15.3 (6.5) mL. In three cases, fetocide was performed by cardiocentesis and in one case lidocaine was unsuccessful and fetocide was performed with KCl. There were no maternal side effects. CONCLUSIONS: Lidocaine is an effective drug to perform fetocide with doses below the toxic dose for the mother. PMID- 12628272 TI - Longitudinal quantification of uterine artery blood volume flow changes during gestation in pregnancies complicated by intrauterine growth restriction. AB - OBJECTIVE: To quantify and compare longitudinal uterine artery volume flow changes in appropriate for gestational age (AGA) pregnancies and those complicated by intrauterine growth restriction (IUGR). DESIGN: Serial longitudinal study. SETTING: Large UK Teaching Hospital Obstetrics and Gynaecology Department and Institute of Anatomy, RWTH Aachen, Germany. POPULATION: Pregnant women with accurately dated singleton pregnancies. METHODS: Quantified uterine volume flow was prospectively measured by colour power angiography in (a) 32 women with abnormal uterine artery Doppler velocimetry at 20 and 24 weeks of gestation and with risk factors for IUGR (IUGR group) and (b) 25 women with normal uterine artery Doppler velocimetry and no risk factors for IUGR (AGA group) between 20 and 38 weeks of gestation. Values obtained from each gestation were compared using unpaired t test. MAIN OUTCOME MEASURES: Gestational age at delivery, birthweight and total quantified volume flow (mL/min) per gestation in IUGR and AGA pregnancies. RESULTS: Twenty of the 32 women recruited into the IUGR group and 18 of the 25 controls fulfilled the criteria for inclusion in the analyses. The mean birthweight [SD] and gestational age [SD] at delivery in the IUGR and AGA groups were 2634 [277] versus 3429 [349] g and 39.5 [1.2] and 41.1 [2.3] weeks, respectively. The diameter of the proximal uterine artery just after it crosses the external iliac artery was smaller in the IUGR group from as early as 20 weeks of gestation but this difference only became statistically significant from 24 weeks of gestation and widened as pregnancy advanced. The quantified volume flow in the IUGR group was significantly less than that in the AGA throughout the study period (287 [117] versus 328 [159] mL/min at 20 weeks (P < 0.05), 334 [169] versus 538 [181] mL/min at 24 weeks [P < 0.004] and 534 [332] versus 830 [284] mL/min at 38 weeks of gestation [P < 0.002]). Volume flow in the IUGR group was 12.5% and 36.7% less than that in the AGA group at 20 and 28 weeks of gestation, respectively. CONCLUSIONS: Proximal uterine artery diameter and quantified volume flow change with gestation and show significant differences between AGA pregnancies and those complicated by IUGR. These changes occur early and become more marked as pregnancy advances. Early use of these measurements may identify pregnancies at risk of complications. PMID- 12628273 TI - Maternal serum activin A levels in association with intrauterine fetal growth restriction. AB - OBJECTIVE: To assess maternal serum activin A as a potential marker of fetal growth restriction. DESIGN: A cohort study. SETTING: A maternal-fetal medicine unit, university teaching hospital. POPULATION: Fifty-seven women with a small fetus (less than 10th centile for gestation) referred for assessment of fetal size by ultrasound biometry. METHODS: At the time of presentation for fetal biometry, maternal blood was collected for activin A measurement. The case records of each woman were independently reviewed after delivery and the pregnancy grouped into one of three groups: constitutionally small fetus, intrauterine growth restricted (IUGR) fetus or IUGR fetus and maternal pre eclampsia (IUGR-pre-eclampsia). Activin A levels in the three groups were compared. MAIN OUTCOME MEASURES: Maternal serum activin A levels. RESULTS: Sixteen of the 57 pregnancies were classified as constitutionally small, 17 as IUGR and 24 as IUGR-pre-eclampsia. Expressed as multiples of a normal median (MoMs), the median (95% CI) activin A level in the constitutionally small pregnancies was 1.12 (0.72-1.39) MoMs significantly lower than the level in both the IUGR pregnancies, 3.00 (1.84-4.11) MoMs, and the IUGR-pre-eclampsia pregnancies, 7.96 (5.73-10.62) MoMs (P = 0.002 and 0.0001 for IUGR vs constitutionally small and IUGR-pre-eclampsia vs constitutionally small, respectively). CONCLUSIONS: Maternal serum activin A may be useful in the assessment of the small for gestational age fetus. PMID- 12628274 TI - Acquired factor VIII inhibitors in pregnancy: data from the Italian Haemophilia Register relevant to clinical practice. AB - OBJECTIVE: To review the clinical problems related to the inhibitor of factor VIII (FVIII) in pregnancy. DESIGN: Retrospective analysis. SETTING: Haemophilia and haemotology centres. POPULATION: Patients registered and followed up at the centres. METHODS: Data were collected from the Italian Haemophilia Register of acquired FVIII inhibitor. RESULTS: Twenty of 96 cases with FVIII inhibitor were identified postpartum. The time of appearance was 3-150 days postpartum. All but one of the cases were idiopathic; 11/20 patients required blood transfusions. In six patients, the inhibitor was identified because of surgical bleeding, four after hysterectomies carried out because of postpartum haemorrhage. A prolonged activated partial thromboplastin time was present in all women in whom the test was carried out. Nine women did not require treatment because the bleeding was mild; in 11 patients bleeding was promptly controlled by different therapeutic modalities. Immunosuppressive therapy was used to suppress the inhibitor. The majority of the patients who achieved complete remission received steroids; in 6/6 patients who relapsed, a second remission was obtained with combined therapy. CONCLUSIONS: In a review of 20 pregnancies with FVIII inhibitor, over a 15 year period, bleeding was controlled in all cases with no fatalities. Correct evaluation of coagulation screening tests, in particular, activated partial thromboplastin time, is essential. PMID- 12628275 TI - Outcomes of pregnancies in women with type 1 diabetes in Scotland: a national population-based study. AB - OBJECTIVE: To determine the outcomes of pregnancies in women with pre-existing, type 1 diabetes. DESIGN: Prospective, population-based cohort. SETTING: Scotland. POPULATION: All 273 women with type 1 diabetes with a pregnancy ending (in miscarriage, abortion or delivery) during the 12 months (from April 1, 1998 to March 31, 1999). METHODS: Pregnancies identified prospectively by clinicians in each hospital; outcome data collected from case records and from Scottish national data sets. MAIN OUTCOME MEASURES: Perinatal and infant mortality, congenital anomaly and birthweight. RESULTS: Of the 273 pregnancies, 40 (14.7%) ended in miscarriage, 20 (7.3%) in abortion and 213 (78%) in delivery. Three deliveries were twin births, thus 216 babies were born. Stillbirth rate (4/216): 18.5 (95% CI 5.1-46.8) per 1000 total births; perinatal mortality rate (6/216): 27.8 (95% CI 10.2-59.4) per 1000 births. There were 13 verified congenital anomalies (in six abortions and seven live births), anomaly rate: 60 (95% CI 32 101) per 1000 total births. Among 208 singleton, live born infants, the mean birthweight was 3427 g. Standardised birthweight scores, relative to a reference population, showed a unimodal distribution, shifted to the right (mean, 1.57 SD). CONCLUSIONS: In an unselected population, adverse outcomes remain more common among the infants of mothers with type 1 diabetes than in the general population. The targets of the St Vincent Declaration of 1989 have not been met. Improvements may be gained by increases in provision of prepregnancy care and in the proportion of pregnancies that are planned. However, further research is needed to clarify the root causes of adverse outcomes in the pregnancies of women with diabetes. PMID- 12628276 TI - Unexpected reduction in the incidence of birth trauma and birth asphyxia related to instrumental deliveries during the study period: was this the Hawthorne effect? AB - OBJECTIVE: The study was originally designed to identify the risk factors that could predict those difficult instrumental deliveries resulting in birth trauma and birth asphyxia. DESIGN: A prospective study on all singleton deliveries in cephalic presentation with an attempt of instrumental delivery over a 12-month period (13 March 2000 to 12 March 2001). SETTING: A local teaching hospital. SAMPLE: Six hundred and seventy deliveries. METHODS: A codesheet was designed to record the demographic data, characteristics of first and second stages of labour and neonatal outcome. In particular, the doctor had to enter the pelvic examination findings before the attempt of instrumental delivery. MAIN OUTCOME MEASURES: Birth trauma and birth asphyxia. RESULTS: There was a significant reduction in the incidence of birth trauma and birth asphyxia related to instrumental deliveries during the study period (0.6%) when compared with that (2.8%) in the pre-study period (1998 and 1999) (RR 0.27, 95% CI 0.11-0.70). There was more trial of instrumental deliveries in the operating theatre although this was not statistically significant (RR 1.19, 95% CI 0.88-1.60). The instrumental delivery rate decreased during the study period (RR 0.88, 95% CI 0.82-0.94). The caesarean section rate for no progress of labour, the incidence of direct second stage caesarean section and the incidence of failed instrumental delivery did not increase during the study period. CONCLUSIONS: Apart from the merits of regular audit exercise and increasing experience of the staff, the Hawthorne effect might be the major contributing factor in the reduction of birth trauma and birth asphyxia related to instrumental deliveries during the study period. PMID- 12628277 TI - Triple compartment prolapse: sacrocolpopexy with anterior and posterior mesh extensions. AB - Post-hysterectomy vault prolapse may be accompanied by anterior (cystocele) and posterior (rectocele) pelvic compartment prolapse. We describe our results with sacrocolpopexy with anterior and posterior polytetrafluoroethylene mesh (SCAPM) extensions. A prospective on-going study is presented of 12 consecutive, complicated patients referred to our tertiary referral unit with a median age of 60 years (range 39-69) who underwent SCAPM between April 1997 and June 1999. All patients had a history and physical examination, International Continence Society Prolapse Staging (ICS) and pre-operative multichannel urodynamics testing with their prolapse reduced. All patients had an ICS grade 2, triple compartment prolapse or worse for inclusion into this study. The operative results were assessed with a 10-point visual analogue scale (VAS) for personal satisfaction and the (non-validated) St George's Hospital symptom questionnaire assessing urgency, urge incontinence, stress incontinence, splinting, digitation, sexual activity and function. All patients were reviewed during February 2002 by an independent observer (B.G. gynaecologist) no longer associated with the Department of Urogynaecology. All patients were followed post-operatively for a median of 39 months (range 32-58). One patient had a recurrent grade 1 cystocele and another a grade 1 rectocele. The median VAS score for personal satisfaction with the operative results was 8 (range 6-10). There was one patient with de novo urgency and one with de novo stress incontinence. Four of the five patients who needed to splint or digitate to empty the rectum no longer required these measures post-operatively. There were no reports of de novo dyspareunia, and two patients had improved vaginal lubrication. Two patients, each with more than five prior vaginal procedures, had a total of three episodes of mesh erosion through the posterior vaginal wall. The SCAPM is an effective treatment for triple compartment prolapse and incomplete rectal emptying. Patient satisfaction is good. PMID- 12628278 TI - Absence of Factor V Leiden, thrombomodulin and prothrombin gene variants in Black South African women with pre-eclampsia and eclampsia. AB - It has been suggested that gene aberrations may contribute to vascular endothelial dysfunction of pre-eclampsia in Caucasian and Japanese women. This study was undertaken to examine the association between pre-eclampsia in Black Zulu speaking South African women and the Factor 5 Leiden mutation. 100 patients with pre-eclampsia comprised the study group. The control group comprised 110 normotensive pregnant women of the same population group. Genotyping was performed to detect the G or A allele at residue 506 of the Factor V gene, and the C or T allele at residue 455 of the thrombomodulin gene. Our findings demonstrate that these particularly genetic loci are of little use in disease association studies for pre-eclampsia in homogenous Zulu speaking Africans. PMID- 12628279 TI - Fracture of the neck of the femur and osteomalacia in pregnancy. PMID- 12628281 TI - Prolapse of the uterine tube after subtotal hysterectomy. PMID- 12628280 TI - Twin pregnancy in each half of a uterus didelphys with a delivery interval of 66 days. PMID- 12628283 TI - Primary malignant follicular lymphoma of the cervix: a rare cause of postmenopausal bleeding. PMID- 12628282 TI - Prenatal diagnosis of fetal bowel obstruction complicated by in utero acute intestinal perforation. PMID- 12628286 TI - [Species identification and molecular epidemiology of bacteria belonging to Ochrobactrum genus]. AB - Two species of medical interest belong to the genus Ochrobactrum, Ochrobactrum anthropi and Ochrobactrum intermedium. They are members of the microbiota of soil and an increasing number of works report the isolation of O. anthropi from clinical specimen, especially from immunocompromised patients and nosocomial infection. Involving of each species in human infection is poorly estimated due to unclear differential phenotypic characters. We performed 16S rDNA sequencing for identification of 20 clinical isolates of Ochrobactrum sp. to the species level. Then, we studied the phenotype of each isolate especially, morphology, culture onto different media and at different temperatures, biochemical characters and antibiotics resistance pattern. Colony morphology after growth onto Trypticase-Soy and McConkey agar, culture at 45 degrees C onto Trypticase Soja agar, presence of urease, and netilmycin, tobramycin and colistin resistance allowed identification of species. Ribotyping using HindIII and EcoRI gave a supplementary criterion for species determination but did not allow typing at the infra-species level. In contrast, Pulsed-Field Gel Electrophoresis showed high degree of polymorphism between strains and proved the clonality of certain isolates. Thus, this method could be a useful tool for molecular epidemiology of Ochrobactrum infections. PMID- 12628285 TI - [A new model of human prostate cancer, the PAC120 xenograft]. AB - Prostate cancer is the second cause of cancer death in men. Often, initialy hormono-independent, escape from anti-androgen therapy is a key event of tumoral progression showing an hormone-independent phenotype. To study morphological, genetic and molecular bases associated with the hormono-dependence escape, a new model of human adenocarcinoma prostate xenograft, PAC120, was established with its hormono-dependent and independent variants. Its growth was strongly inhibited by surgical castration or by administration of the new gonadotrophin-releasing hormone antagonist, FE 200486 (Ferring, San Diego, CA). Evolution to hormono independence was frequently associated with a mucoid differentiation or a neuroendocrine-like pattern, with the apparition of new chromosomic alterations and variations of human gene expressions. PAC120 xenograft is a new model of hormone-dependent prostate cancer, opening the opportunity to study the hormone dependence escape mechanism and to evaluate the efficacity of new therapeutics. PMID- 12628287 TI - [Epidemiological study of tuberculosis in the area of Angers, France, as studied by 3 PCR-based fingerprinting methods]. AB - The new genotyping methods efficiently complement classical epidemiological investigation in order to attempt a global approach to TB control. In the present work, we have studied the genomic diversity of Mycobacterium tuberculosis isolated during the year 1998 within the district of Angers, France (260,000 inhabitants distributed in 29 districts), in order to identify recent transmission events and any related risk factors. The methods used included "spacer oligonucleotide typing" or spoligotyping, "variable number of DNA tandem repeats" or VNTR, and "double repetitive element PCR" or DRE-PCR. The resulting spoligotyping and VNTR results were also feeded to international databases and compared with >10,000 isolates for spoligotyping and 500 isolates for VNTR, representative of about 60 countries. The results obtained underlined that most of the TB cases in our setting probably reflected reactivation cases, as clustered cases indicative of potential events of recent transmission were rare. Furthermore, interrogation of international databases showed that most of the isolates from the Angers region belonged to major conserved families of TB isolates representative of Europe, with only rare cases of Asian origin, or those previously reported in specific epidemies reported from elsewhere. PMID- 12628288 TI - [Antibiotic resistance of Staphylococcus aureus at north Lebanon: place of the methicillin resistance and comparison of detection methods]. AB - The aim of this study was to evaluate the susceptibility of 100 Staphylococcus aureus strains isolated from the laboratory of Microbiology of the Islami Hospital of Tripoli (Lebanon) to 19 antibiotics, and to determine the prevalence of methicillin resistant strains. 30% of strains studied were methicillin resistant, 96% were resistant to the penicillin G. Clavulanic acid restaurated the amoxicillin activity to 29%. The resistance level was 34% for amikacin, 3% for gentamycin and tobramycin, 10% for chloramphenicol, 44.33% for tetracyclin, 7% for erythromycin, 4.04% for clindamycin, 20% for trimethoprim-sulfametoxasol and 0% for vancomycin and teicoplanin. The methicillin-resistant Staphylococcus aureus possess more important resistant level in comparison with the methicillin sensitive strains. We compared the ability of latex agglutination test (Slidex(R) SARM, bioMerieux, France) to detect the production of penicillin-binding protein 2' (PBP 2') in 100 clinical isolates of S. aureus with two reference methods: the oxacillin disk diffusion test and the MIC determination by the E-test (AB BIODISK, Sweden). The two reference methods give the same results for the detection of methicillin resistant S. aureus. The Slidex test was positive for all 30 isolates determined to be methicillin resistant by the reference methods (sensitivity 100%). The latex test was negative for 42 of 70 isolates determined to be methicillin susceptible by the reference methods, and the latex test was positive for 28 isolates determined to be susceptible (specificity 60%). PMID- 12628289 TI - [Molecular basis of the t(1;22)(p13;q13) specific for human acute megakaryoblastic leukemia]. AB - The t(1;22)(p13;q13) translocation is specifically associated with infant acute megakaryoblastic leukemia (M7). We have recently characterized the two genes involved in this translocation: OTT (One Two Two) and MAL (Megakaryoblastic Acute Leukemia) respectively located on chromosome 1 and 22. The t(1;22) translocation results in the fusion of these genes in all the cases studied to date. We summarize here present knowledge regarding this translocation. PMID- 12628290 TI - [The stathmin-tubulin interaction and the regulation of the microtubule assembly]. AB - Stathmin family proteins interact with tubulin and negatively regulate its assembly in microtubules. One stathmin molecule forms a complex with two alphabeta tubulin heterodimers in an interaction that is weakened upon stathmin phosphorylation. The X-ray structure of crystals of the complex reveals a head-to tail arrangement of the two tubulins which are connected by a long stathmin alpha helix. By holding tubulins in a curved complex that is not incorporated in microtubules, stathmin lowers the pool of "assembly competent" tubulin. An alternate mechanism has been also proposed to account for the stathmin action in vivo; it involves a direct interaction of stathmin with microtubule (+) ends. More experiments are needed to evaluate the relative contribution of this alternative mechanism to the regulation of tubulin assembly by stathmin. PMID- 12628291 TI - [Myocardial microdialysis. Importance and potential in cardiovascular research]. AB - The microdialysis expanded mainly in the field of the neuro- and the dermopharmacology with the study of the transmitters released in the central nervous system and derm. Since ten years, this tool gained other disciplines such as cardiology and cardiovascular surgery. Indeed, the collection and the study of the molecules released in the myocardic interstitial fluid without deteriorating it functioning made microdialysis a powerful tool in the study of the extracellular environment of the cardiomyocyte. The purpose of this study is to point out the principle of the microdialysis and to show its various uses in the field of cardiovascular pharmacology. PMID- 12628292 TI - [Diagnosis of imported malaria with multiplex PCR on LighCycler apparatus]. AB - For the diagnosis of imported malaria, a competitive multiplex PCR using LightCycler was developed and compared for 3 months with a traditional PCR method. Hundred eighty three patients were tested by these 2 techniques of molecular diagnosis: 60 were positive for P. falciparum, 9 were positive for other Plasmodium species and 114 were negative. The LightCycler method was found in total agreement with the traditional PCR. PMID- 12628293 TI - [Premature ovarian failure in galactosaemia: pathophysiology and clinical management]. AB - Classic galactosaemia is a rare aetiology of premature ovarian failure. It is caused by galactose-1-phosphate uridyltransferase deficiency and leads to a severe disease in the newborn. This acute toxic syndrome will completely regress under a galactose-free diet, but some long-term complications, particularly hypergonadotropic hypogonadism in female patients, are frequently observed. Ovarian toxicity could be due to intracellular accumulation of galactose metabolites or to deficient glycosylation reactions. Moreover, the tremendous follicular decrease in the galactosaemic ovary could also involve programmed cell death (apoptosis). As the exact mechanisms of this ovarian injury are still unknown, there is no prevention of follicular loss, thus clinical management especially includes hormonal replacement therapy in order to prevent bone loss and cardiovascular risks and sometimes to allow patients to become pregnant. PMID- 12628294 TI - Protection against experimental P falciparum malaria is associated with short AMA 1 peptide analogue alpha-helical structures. AB - Apical membrane antigen-1 (AMA-1) is an integral Plasmodium falciparum malaria parasite membrane protein. Peptides having high activity binding to human red blood cells have been identified in this protein. One of them, peptide 4325, with the amino acid sequence MIKSAFLPTGAFKADRYKSH, for which critical binding residues have already been defined (underlined), is conserved and non-immunogenic. Its critical binding residues were changed for amino acids having similar mass but different charge to change such immunological properties. These changes rendered some peptides immunogenic and protective against experimental challenge in Aotus monkeys. Three-dimensional models of peptide 4325 and its analogues, 20032 and 20034, were calculated from NMR experiments with distance geometry and restrained molecular dynamic methods. Non-immunogenic, non-protective peptide 4325 showed differences in its secondary structure with respect to protective, immunogenic peptides 20032 and 20034. Such data suggest that these modifications could have converted non-immunogenic peptides into immunogenic, protective ones, making them excellent candidates for a multi-component subunit synthetic malaria vaccine. PMID- 12628295 TI - The respiratory chain complex thresholds in mitochondria of a Drosophila subobscura mutant strain. AB - Analysis of a mutant strain of Drosophila subobscura revealed that most (80%) mitochondrial genomes have undergone a large scale deletion (5 kb) in the coding region. Compared with the wild-type strain, complex I and III activities are, respectively, reduced by 50% and 30% in the mutant. However, the ATP synthesis capacities remain unchanged. In order to elucidate how the ATP synthesis is maintained at a normal level, despite a significant decrease in complex I and III activities, we progressively inhibited respiratory chain complex activities, respiration rate and ATP synthesis. Complex I, III and IV activities were inhibited by rotenone, antimycin and KCN, respectively. Threshold curves were thus determined for each complex. Our results demonstrated that in the mutant strain, both mitochondrial respiration and ATP synthesis had decreased when complex I activity was inhibited by more than 20%, whereas 70% inhibition is required to induce similar changes in the wild-type. The complex I inhibition pattern of the wild-type was restored by a backcross (mutant female/wild-type male). The complex III activity threshold is below 20% in both strains, and we observed some difference in antimycin sensitivity, suggesting a modification of the complex enzymatic properties in the mutant. In contrast, threshold values of 70% were measured for complex IV inhibition. Our data suggest that the difference in the complex I threshold curves between the wild-type and mutant strains could partially account for the absence of pathological phenotype in the mutant. PMID- 12628296 TI - Lipid peroxidation-mediated cytotoxicity and DNA damage in U937 cells. AB - Membrane lipid peroxidation processes yield products that may react with DNA and proteins to cause oxidative modifications. In the present study, we evaluated lipid peroxidation-mediated cytotoxicity and oxidative DNA damage in U937 cells. Upon exposure of U937 cells to tert-butylhydroperoxide (t-BOOH) and 2,2'-azobis (2-amidinopropane) hydrochloride (AAPH), which induce lipid peroxidation in membranes, the cells exhibited a reduction in viability and an increase in the endogenous production of reactive oxygen species (ROS), as measured by the oxidation of 2',7'-dichlorodihydrofluorescein. In addition, a significant decrease in the intracellular GSH level and the activities of major antioxidant enzymes were observed. We also observed lipid peroxidation-mediated oxidative DNA damage, reflected by an increase in 8-OH-dG level and loss of the ability of DNA to renature. When the cells were pretreated with the antioxidant N-acetylcysteine (NAC) or the spin trap alpha-phenyl-N-t-butylnitrone (PBN), lipid peroxidation mediated cytotoxicity in U937 cells was protected. This effect seems to be due to the ability of NAC and PBN to reduce ROS generation induced by lipid peroxidation. These results suggest that lipid peroxidation resulted in a pro oxidant condition of U937 cells by the depletion of GSH and inactivation of antioxidant enzymes, which consequently leads to a decrease in survival and oxidative damage to DNA. The results indicate that the peroxidation of lipid is probably one of the important intermediary events in oxidative stress-induced cellular damage. PMID- 12628297 TI - Prohibitin and prohibitone are contained in high-molecular weight complexes and interact with alpha-actinin and annexin A2. AB - The closely related proteins prohibitin (p32) and prohibitone (p37) are evolutionarily conserved with homologues found from cyanobacteria to man. They are thought to be exclusively mitochondrial and have been assigned many-rather different-functions, ranging from a role in lifespan, in mitochondrial inheritance and as chaperones of mitochondrial proteases in yeast. Evidence for a localisation outside of mitochondria has been brought forward in mammalian cells, where they influence cell-cycle progression and are found in association with cell surface receptors. We have employed a yeast two-hybrid screen to identify other interacting proteins and have identified alpha-actinin and annexin A2 as binding partners for prohibitin and prohibitone. Coprecipitation experiments supported the putative binding between prohibitin and prohibitone on the one hand and annexin A2 or alpha-actinin on the other hand in intact cells. Surface plasmon resonance analysis was used to determine relative affinities between prohibitin and alpha-actinin and between prohibitone and annexin A2 and alpha actinin, respectively. We further show that prohibitin and prohibitone can also form homomeric (preferentially tetrameric) and heteromultimeric complexes, with significant affinities. PMID- 12628298 TI - Cloning of the pig renal organic anion transporter 1 (pOAT1). AB - A pig kidney cDNA library was screened for the porcine ortholog of the multispecific organic anion transporter 1 (pOAT1). Several positive clones were isolated resulting in two alternatively spliced cDNA clones of pOAT1 (pOAT1 and pOAT1A). pOAT1-cDNAs consist of 2126 or 1895 base pairs (EMBL Acc. No. AJ308234 and AJ308235) encoding 547 or 533 amino acid residue proteins with 89, 87, 83 and 81% homology to the human, rabbit, rat, and mouse OAT1, respectively. Heterologous expression of pOAT1 in Xenopus laevis oocytes revealed an apparent K(m) for [3H]PAH of 3.75 +/- 1.6 microM. [3H]PAH uptake mediated by pOAT1 was abolished by 0.5 mM glutarate or 1 mM probenecid. Functional characterization of pOAT1A did not show any affinity for [3H]PAH. In summary, we cloned two alternative splice variants of the pig ortholog of organic anion transporter 1. One splice form (pOAT1) showed typical functional characteristics of organic anion transporter 1, whereas the second form appears not to transport PAH. PMID- 12628299 TI - Immunological studies of sheep brain keratan sulphate proteoglycans. AB - Recently, we reported the isolation and partial characterization of keratan sulphate (KS) from sheep brain. In this study, a panel of monoclonal antibodies (Mab) recognizing epitopes within KS chains and core proteins of KS-containing proteoglycans were used to detect, by immunoblotting, antigenically related molecules extracted from cerebrum, cerebellum and brainstem, respectively. Although the intensity of labelling varied with each of the antibodies, the brain KSPGs were recognized by all the monoclonals used, confirming the presence of KS side chains, which react with the Mabs: 5-D-4, EFG-11, EFG-4, I22, as also the presence of KSPGs related to phosphacan-KS (3H1 proteoglycan). Extracts of all the three brain areas could bind both anti-KS and anti-core protein Mabs, as also anti-HNK-1 monoclonal antibody. Binding was sensitive to keratanases degradation in the cerebrum and brainstem except cerebellum where the presence of a large molecular size hybrid CS/KSPG bearing KS chains partially resistant to keratanases was identified. This population reacts only with 5-D-4, EFG-11 and EFG-4 antibodies. Furthermore, the presence of HNK-1 epitope in CSPGs was detected in the cerebellum and brainstem. In contrast, in the cerebrum the coexistence of HNK-1 epitope and KS in KSPGs was identified. These data suggest that the KSs of sheep brain are part of proteoglycans containing protein and KS antigenic sites related to those of corneal and cartilage KSPG, as also of the brain proteoglycan phosphacan-KS. PMID- 12628300 TI - Poly(ADP-ribose) degradation by post-nuclear extracts from human cells. AB - The nuclear metabolism of poly(ADP-ribose) is mainly regulated by poly(ADP ribose) polymerase-1 (PARP-1) and by poly(ADP-ribose) glycohydrolase (PARG). A PARP-like enzyme, V-PARP, and a PARG isoform are present in the extra-nuclear compartment of mammalian cells, even if poly(ADP-ribose) has never been detected therein. In this work, we demonstrate the ability of post-nuclear extracts from HeLa and HL60 cells to degrade synthetic 32P-polymers of ADP-ribose to ADP-ribose and, further, to AMP. This reaction implies the combined action of PARG and of an ADP-ribose-degrading activity, possibly corresponding to a phosphodiesterase and/or to an ADP-ribose pyrophosphatase. The inhibition of PARG or ADP-ribose degrading enzymes allowed the demonstration that in vitro synthesized 32P poly(ADP-ribose) is first digested to ADP-ribose monomers by a typical PARG reaction, and that ADP-ribose is further rapidly converted into AMP by an Mg(2+) dependent activity. Collectively, our results demonstrate the ability of the human cell post-nuclear fraction to convert synthetic poly(ADP-ribose) into utilizable AMP units by the concerted action of PARG and ADP-ribose-degrading activities. PMID- 12628301 TI - Occurrence and structural characterization of versican-like proteoglycan in human vitreous. AB - Human vitreous gel is a special type of extracellular matrix, in which interpenetrating networks of collagen fibrils and hyaluronan are found. In this study, we report that apart from significant amounts of collagen, hyaluronan and sialylated glycoproteins, it was found that the human vitreous gel also contained low amounts of versican-like proteoglycan. The concentration of versican-like proteoglycan in the whole vitreous is 0.06 mg protein/ml of vitreous gel and represents a small percentage (about 5%) of the total protein content. The versican-like proteoglycan has a molecular mass of 380 kDa, as estimated by gel chromatography. Its core protein is substituted by chondroitin sulphate side chains (average molecular weight 37 kDa), in which 6-sulphated disaccharides predominated. According to the physicochemical data, the number of chondroitin sulphate chains is likely to be 5-7 per molecule. These proteoglycan monomers form large aggregates with endogenous hyaluronan. Versican, which is able to bind lectins via its C-terminal region, may bridge or interconnect various constituents of the extracellular matrix via its terminal domains in order to stabilize large supramolecular complexes at the vitreous, contributing towards the integrity and specific properties of the tissue. PMID- 12628302 TI - Purification of classical pancreatic lipase from dog pancreas. AB - The purification of canine classical pancreatic lipase from canine pancreatic juice, but not from pancreatic tissue, has been reported previously. Given the logistic difficulties associated with collection of pancreatic juice in dogs and efforts to minimize experiments in live animals the objective of this project was to purify canine classical pancreatic lipase from dog pancreas. Dog pancreata were collected from research dogs that had been sacrificed for unrelated research projects. Pancreatic tissue was delipidated using organic solvents. The delipidated pancreatic extract was further purified by extracting the enzymes in a Tris-buffer containing two different protease inhibitors, benzamindine and phenylmethylsulfonyl fluoride (PMSF), followed by anion exchange chromatography, gel-filtration, and cation exchange chromatography. The purified protein showed a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis with a molecular weight of approximately 50.7. Isoelectric focusing showed isoelectric points ranging from 6.0 to 6.2. N-terminal amino acid sequencing of the first 25 amino acid residues showed the sequence Lys-Glu-Val-X-Phe-Pro-Arg-Leu-Gly-X-Phe Ser-Asp-Asp-Ser-Pro-Trp-Ala-Gly-Ile-Val-Glu-Arg-Pro-Leu. This sequence showed close homology with classical pancreatic lipase in pigs, horses, and human beings. We conclude that canine classical pancreatic lipase can be successfully purified from canine pancreatic tissue. PMID- 12628303 TI - A reporter gene assay for evaluation of tissue-specific responses to estrogens based on the differential use of promoters A to F of the human estrogen receptor alpha gene. AB - INTRODUCTION: Reporter gene assays are useful means for monitoring cellular responses. We report here a reporter gene assay for evaluating and monitoring estrogen activities by estrogen-like compounds and xenoestrogens, which is based on the promoters from the human estrogen receptor alpha (ERalpha) gene. METHODS: The reporter gene constructs contained a proximal promoter region (containing promoters A and B: ProAB) or either of promoters C to F (ProC, ProD, ProE, and ProF) or fused minor promoters (ProCDEF). These constructs were first used to evaluate promoter activity in cell lines derived from breast (MCF-7 and T-47D cells), ovary (SK-OV-3 cells), endometrium (Ishikawa cells), and stomach (MKN-28 cells). RESULTS: Besides very high levels of activity by ProAB in all of the cell lines tested, moderate levels were detected for ProD in the breast and endometrium cell lines and for ProF in the ovary and endometrium cell lines. A moderate level of activity by ProE was detected only in the stomach cells. Differences in estrogen-like activity between ProAB and ProD were observed for tamoxifen and bisphenol A (BPA) in MCF-7 cells. DISCUSSION: The assay proposed here might provide expression profiles of cancer cells of various origins for evaluating the estrogen responsiveness and for identifying tissue- or cancer cell specific transcription factors. PMID- 12628304 TI - Development of a high-throughput screening-amenable assay for human poly(ADP ribose) polymerase inhibitors. AB - INTRODUCTION: Poly(ADP-ribose) polymerase (PARP) plays a pivotal role in the repair of DNA strand breaks. However, excessive activation of PARP causes a rapid depletion of intracellular energy, leading to cell death. Inhibitors of PARP have been shown to reduce infarct size in animal models of myocardial ischemia. PARP inhibitors may have potential therapeutic benefit in the treatment of myocardial ischemia, stroke, head trauma, and neurodegenerative disease, and as an adjunct therapy with chemotherapeutic agents/radiation in cancer therapy. METHODS: Assays reported in the literature and commercially available PARP assay kits are labor intensive, use radioactive reagents, use antibodies, and are not readily amenable to high throughput screening (HTS) [corrected]. Here we report the development and the validation of a nonradioactive PARP assay suitable for HTS. This is a biotinylated NAD-based colorimetric assay in a 96-well plate format. RESULTS: The assay is sensitive, reproducible, and easy to use. The IC(50) values generated for the known PARP inhibitors are in agreement with those generated using the commercial radioactive kit and those reported in the literature. DISCUSSION: The present study demonstrates a sensitive and reproducible methodology capable of screening human PARP inhibitors in high-throughput format. PMID- 12628305 TI - A high-throughput cell-based reporter gene system for measurement of CYP1A1 induction. AB - INTRODUCTION: Enzyme induction is undesirable in new drug discovery process, with consequences spanning from auto-induction to toxicity. Cytochrome P450 (CYP) 1A1 has long been known to be one of the metabolic enzymes involved in activating many procarcinogens, the first step toward tumor formation during chemical carcinogenesis. Induction of CYP1A1 during drug treatment may predispose the patients to some risk of chemical carcinogenesis. METHODS: Based on the signal transduction mechanism of CYP1A1 induction, a high-throughput reporter-gene system was established by stable transformation of H4IIE cells to incorporate the luciferase gene under control of CYP1A1 promoter. This stable cell line was validated with known CYP1A1 inducers, such as 3-methylcholanthrene (3-MC), beta naphthoflavone (beta-NF), alpha-naphthoflavone (alpha-NF) and 3-indocarbinol. Thirty in-house new chemical entities (NCEs) were then screened with this reporter-gene system, and also administered to rats to evaluate in vivo CYP1A1 induction. RESULTS: CYP1A1 reporter gene system can be used to identify strong inducers, such as 3-MC, beta-NF and alpha-NF, and weak inducers, such as 3 indocarbinol. In vitro induction of 30 in-house compounds in reporter gene system did not correlate with in vivo induction in rat liver microsome measured by ethoxyresorufin-O-dealkylation (EROD) activity, but had a reasonable correlation with Western blot signals. DISCUSSION: This reporter-gene system may be useful in eliminating compounds that can cause CYP1A1 induction at an early stage of drug discovery. PMID- 12628307 TI - Biosynthesis of drug glucuronides for use as authentic standards. AB - INTRODUCTION: Glucuronidation by the uridine diphosphate glucuronosyltransferases (UGTs) plays a pivotal role in the clearance mechanism of both xenobiotics and endobiotics. The detection of glucuronides at low micromolar concentrations is required to accurately model in vitro enzyme kinetics and in vivo pharmacokinetics. However, relatively few glucuronides are currently available as standards for developing liquid chromatography and mass spectroscopy (LC/MS) bioanalytical methods. METHODS: The glucuronidation capacity of hepatic microsomes prepared from rat (RLM), dog (DLM), monkey (MLM), and human (HLM) was examined for five xenobiotic substrates. In each case, glucuronide standards were produced using the enzyme source most efficient for the production of that specific glucuronide. RESULTS: Dog hepatic microsomes were used to produce glucuronides for anthraflavic acid (yield: 14 mg), buprenorphine (yield: 14 mg), and octylgallate (total yield: 13 mg), whereas propofol glucuronide (yield: 20 mg), and ethinylestradiol glucuronide (yield: 8 mg) were prepared using HLM. All glucuronides were characterized by LC/MS/MS and nuclear magnetic resonance (NMR) spectroscopy. DISCUSSION: The multimilligram quantities of glucuronide standards produced by this method have many applications throughout drug discovery and toxicology. In addition to allowing the quantification of glucuronide formation from in vitro and in vivo studies, the authentic standards produced could also be used to assess potential pharmacological or toxicological effects of metabolites. PMID- 12628306 TI - Renal medullary interstitial infusion is a flawed technique for examining vasodilator mechanisms in anesthetized rabbits. AB - INTRODUCTION: In rats, medullary interstitial (IMI) infusion is a useful technique for selective delivery of pharmacological agents to the renal medulla, in both acute and chronic experimental settings. We examined the feasibility of using this technique for delivery of vasodilators in rabbits, since this larger species would provide a number of advantages, particularly in long-term studies of circulatory control. METHODS: Rabbits were anesthetized with pentobarbitone and artificially ventilated. Catheters were placed in a side branch of the renal artery and/or the renal medullary interstitium. Renal blood flow (RBF) was determined by transit-time ultrasound flowmetry, and blood flow in the cortex and medulla was estimated by laser Doppler flowmetry. RESULTS: Pilot studies showed that renal arterial (IRA) infusions of bradykinin (10-300 ng/kg/min) and adenosine (1-10 ng/kg/min) produced only transient renal vasodilatation. IRA infusions of methylamine hexamethylene methylamine (MAHMA) NONOate (100-1000 ng/kg/min) and acetylcholine (10-250 ng/kg/min) produced dose-dependent and sustained increases in RBF and reductions in arterial pressure at the highest doses. However, IMI infusion of the same doses did not consistently increase medullary laser Doppler flux (MLDF). After IRA MAHMA NONOate and IMI acetylcholine, RBF fell to below its resting level. IRA boluses of acetylcholine (10-1250 ng/kg), bradykinin (2-250 ng/kg), and MAHMA NONOate (100-3000 ng/kg) dose-dependently increased RBF and CLDF and MLDF. DISCUSSION: We had previously validated the IMI infusion technique for intramedullary delivery of vasoconstrictors in rabbits. Our present results indicate that this technique has limited application for delivery of vasodilator agents, in part because counterregulatory vasoconstrictor mechanisms are activated. PMID- 12628308 TI - An ELISA method for detection of human antibodies to an immunotoxin. AB - INTRODUCTION: The use of biological molecules, such as immunotoxins, as pharmaceuticals is limited by the presence and development of human antibodies to these agents. This immune response can cause significant inflammatory-related toxicities and can interfere with the efficacy of the biological agent. Therefore, a clinically applicable method to detect these human antibodies is needed for screening patients prior to enrollment and for monitoring patients during treatment. The SS1(dsFv)-PE38 immunotoxin currently in clinical trials is a hybrid molecule targeted against mesothelin-expressing cancer cells via the Fv portion of a murine antibody linked to the Pseudomonas exotoxin (PE), which can inhibit protein synthesis leading to cell death. The objective of this study was to determine if an enzyme-linked immunosorbent assay (ELISA)-based method could be used to detect human anti-SS1(dsFv)-PE38 antibodies in patient serum. METHODS: Human antibodies to the immunotoxin in serially diluted serum specimens were captured on immunotoxin-coated ELISA plates, and detected using a secondary goat antihuman antibody linked to biotin in combination with horseradish peroxidase linked to avidin D (HRP-Avidin). The color was developed with tetramethyl benzidine (TMB). Curves of optical density (OD(630)) versus dilution for 44 serum specimens were compared with positive and negative control serum specimens to classify the serum as positive or negative for anti-immunotoxin antibodies. RESULTS: Ten out of the 40 patients screened were positive for anti-immunotoxin antibodies. Repeated testing of seven samples produced the same results in two independent experiments. The first two patients treated with the immunotoxin developed anti-immunotoxin antibodies during treatment. The results were in perfect concordance with a tissue culture-based neutralization assay performed by an independent laboratory. DISCUSSION: An ELISA-based strategy using an immunotoxin to capture human anti-immunotoxin antibodies provides a consistently accurate technology for screening and monitoring patient serum specimens in clinical trials. PMID- 12628310 TI - The development and testing of a dermal exposure system for pharmacokinetic studies of administered and ambient water contaminants. AB - INTRODUCTION: In order to investigate the pharmacokinetics of water-borne chemicals while eliminating exposures by other routes, a dermal exposure system was developed to expose the hand and forearm of human subjects. METHODS: The goal was, primarily, to study the dermal pharmacokinetics of methyl tertiary butyl ether (MTBE), a water contaminant, and, secondarily, the ambient disinfection byproducts (DBPs). MTBE is used as a fuel oxygenate and DBPs result from chlorination of drinking water. The DBPs measured in the water and blood of the subjects were chloroform, bromodichloromethane, and dibromochloromethane. The dermal exposure system was constructed of inert and impervious materials. The interface between the glass and Teflon exposure tank and the subject was custom made of clear Tedlar (polyvinylfluoride) so that the depth of the arm in the media could be monitored. RESULTS: Sampling of the water concentration of the test chemical, MTBE, demonstrated stability over the duration of the exposure. A temperature loss of about 1.5 degrees C occurred over the course of the 1-h exposure. Blood concentrations taken from 14 human subjects before, during, and after the 1-h exposure demonstrated that measurable MTBE and DBPs were absorbed. DISCUSSION: This system has the advantages of maintaining contaminant concentration and exposing an anatomically distinct body region, and the convenience of blood sampling. PMID- 12628309 TI - Use of dominant negative nuclear receptors to study xenobiotic-inducible gene expression in primary cultured hepatocytes. AB - INTRODUCTION: To determine the feasibility of using dominant negative nuclear receptors to dissect the regulation of inducible gene expression in primary cultured hepatocytes, a series of dominant negative nuclear receptor expression plasmids were designed with truncated AF-2 subdomains. METHODS: Plasmids expressing dominant negative or wild-type constitutive androstane receptor (CAR), pregnane X receptor (PXR), farnesoid X receptor (FXR), liver X receptor (LXR), or peroxisome proliferator-activated receptor alpha (PPARalpha) were transiently cotransfected into primary cultured rat hepatocytes, together with an appropriate reporter plasmid. RESULTS: Treatment with prototypic inducers, 10(-4) M phenobarbital (CAR activator), 10(-5) M pregnenolone 16alpha-carbonitrile (PXR activator), 3x10(-5) M chenodeoxycholate (FXR activator), or 10(-4) M ciprofibrate (PPARalpha activator), significantly activated expression from the corresponding reporter plasmid. Treatment with 22(R)-hydroxycholesterol (LXR activator) only weakly activated the LXR-responsive reporter, while pregnenolone 16alpha-carbonitrile treatment significantly activated this reporter. Cotransfection with wild-type LXRalpha strongly enhanced 22(R)-hydroxycholesterol inducible expression from the LXR-responsive reporter. Cotransfection of hepatocyte cultures with each of the dominant negative nuclear receptor plasmids significantly inhibited inducible expression of the corresponding reporter while, with one exception (LXRalpha), cotransfection with the wild-type receptor moderately enhanced or had little effect on reporter expression. When each dominant negative nuclear receptor was cross-examined against all inducer reporter pairs, effects on multiple inducer-reporter pairs were frequently observed. However, in general, only cotransfection with the appropriate dominant negative inhibited inducible reporter expression to a greater extent than did cotransfection with the corresponding wild-type receptor. DISCUSSION: We suggest that the application of dominant negative nuclear receptors has utility in transient transfection studies aimed at discerning the regulatory role of individual nuclear receptor transcription factors in inducible hepatic gene expression, provided that appropriate controls are employed. PMID- 12628311 TI - Activation of alpha(1)-adrenergic receptors potentiates the nephrotoxicity of ethylene dibromide. AB - Ethylene dibromide (EDB) has been used as a model compound for eliciting hepato- and nephrotoxicity. Conjugation with glutathione (GSH) has been shown to play a role in the bioactivation of EDB. The aim of this study was to determine whether activation of alpha(1)-adrenergic receptors, which causes a decrease in cellular GSH levels, could modulate the nephrotoxicity of EDB. For this purpose, male ICR mice were treated with EDB and/or the alpha-adrenergic agonist, phenylephrine (Pe), or the alpha-adrenergic antagonist, phentolamine (Phe). Animals treated with EDB (40 mg/kg, i.p.) had a 9.3-fold increase in urinary gamma glutamyltranspeptidase (GGTP: EC 2.3.2.2) activity and a 38% decrease in renal non-protein bound sulfhydryl (NPSH) levels; however, animals co-treated with EDB and Pe (50 mg/kg, i.p.) exhibited a 27.8-fold increase in urinary GGTP activity and a 60% decrease in NPSH levels. The enhanced presence of urinary GGTP and decrease in cellular levels of NPSH was nearly blocked by treating animals concomitantly with EDB and Phe (10 mg/kg, i.p.) or EDB, Pe, and Phe. Histopathological examination revealed the enhanced degree of tissue damage and necrosis following treatment with EDB and Pe, and the protective effect of Phe at ameliorating EDB toxicity. These results indicate that factors that can influence alpha-adrenergic receptors may be critical in assessing dose-response data used in the risk assessment process. PMID- 12628312 TI - Receptor-mediated endocytosis of trichosanthin in choriocarcinoma cells. AB - Trichosanthin (TCS) is a ribosome inactivating protein (RIP). It is generally believed that its many biological activities act through inhibition of ribosomes resulting in a decrease in protein synthesis. It has been hypothesized that the rate of entry of TCS into cells to reach ribosomes is an important factor in determining its biological activity. To prove this hypothesis, we have mapped out and compared the intracellular routing of TCS in two cell lines, namely the choriocarcinoma JAR cell line, which is known to be highly sensitive to the toxic effects of TCS, and the hepatoma H35 cell line, to which TCS shows minimal toxicity. Results from laser scanning confocal microscopy indicated that fluorescein isothiocyanate labeled TCS quickly accumulated inside JAR cells within 4 h of incubation while only a low level of fluorescent signals was detected in H35 cells during the same period of time. When TCS was conjugated with gold particles (Au) and its intracellular locations were traced with a transmission electron microscope, it was found that most of TCS were bound to coated pits on the JAR cell surface and were rapidly internalized within an hour. By 4 h, TCS reached almost every cytoplasmic region including ribosomes, and the JAR cell began to degenerate. In H35 cells, however, the binding of TCS to coated pits was not observed, but instead, a small amount of TCS was found to penetrate the cell non-specifically by direct diffusion across the cell membrane. Our observations suggest that most of TCS enter JAR cells via a specific receptor mediated pathway, which allows a swift transport of TCS across the membrane and a rapid accumulation of intracellular TCS, while in H35 cells, TCS takes a slow and non-specific route. The receptor-mediated uptake together with the specific intracellular routing of TCS may partly account for the differential vulnerability of the choriocarcinoma cell line towards the toxicity of TCS. PMID- 12628313 TI - Pulmonary toxicity of thuringiensin administered intratracheally in Sprague Dawley rats. AB - The purpose of this work is to evaluate the pulmonary toxicity of purified thuringiensin in Sprague-Dawley rats. Rats were intratracheally instillated with 0, 0.4, 0.8, 1.6, 3.2, 6.4 and 9.6 mg/kg body weight of thuringiensin. The results indicated that the acute pulmonary LD(50) of thuringiensin for rats was 4.4 mg/kg. The total number of inflammatory cells and lactate dehydrogenase (LDH) activity in the bronchoalveolar lavage (BAL) fluid increased in a dose-dependent manner after thuringiensin instillation. Furthermore, an effective dose of 1.6 mg/kg was selected for the time course study of pulmonary toxicity. The treated animals showed a significant increase in the weights of the lungs, hydroxyproline levels in the lungs and total number of cells in BAL fluid 2, 4, 7, 14, 28 and 56 days after treatment. In comparison with the control, the total protein concentrations in BAL fluid were increased by 361, 615, 116, 41, 34 and 41%, after 2, 4, 7, 14, 28 and 56 days, respectively. The LDH activity in BAL fluid showed a significant increase after 1, 2, 4, 7, 14, 28 and 56 days. The increases in fibronectin levels were 164, 552, 490, 769, 335, 257 and 61% at the corresponding times, but neither tumor necrosis factor nor interleukin-1 increased. The treated rats presented abnormal histology including distributed inflammation in the bronchioles and alveoli, bronchial cellular necrosis on days 1 and 2, and areas of septal thickening with cellular infiltration and collagen deposit in the intestinal and alveolar spaces on days 4-56. Based on these biochemical and pathological parameters, intratracheal instillation of purified thuringiensin might cause significant pulmonary toxicity in rats. PMID- 12628314 TI - Violacein and its beta-cyclodextrin complexes induce apoptosis and differentiation in HL60 cells. AB - Violacein, a pigment isolated from Chromobacterium violaceum, has been reported to have multiple biological activities including in vitro antitumor effects. Certain anticancer agents are known to induce apoptosis in human tumor cell lines. In this work, our aim was to investigate the effectiveness of violacein/beta-cyclodextrin (beta-CD)-containing systems to produce lethal effects in the human promyelocytic leukemia cell line HL60. Using the MTT tetrazolium reduction test, IC(50) for the inclusion complexes (1:1 and 1:2 violacein:beta-CD molar ratios) and violacein alone were less than 1 microM. Violacein and violacein/beta-CD complexes were able to induce NBT reduction. Moreover, by using the Feulgen reaction, all the compounds were found to trigger apoptosis in HL60 cells, inducing around 35% of DNA fragmentation, as analyzed through the diphenylamine assay. In addition, caspases seem to play an important role in the activation of the executioner phase of apoptosis induced by violacein and its derivatives. PMID- 12628315 TI - Lack of estrogenic or (anti-)androgenic effects of d-phenothrin in the uterotrophic and Hershberger assays. AB - Synthetic pyrethroids are among the most common insecticides and pesticides currently in use worldwide. Recently, d-phenothrin, a synthetic pyrethroid, is suspected to have endocrine activities through the estrogen and androgen receptors. However, no study has been conducted to evaluate its potential for hormonal activity using an in vivo test specifically focused on estrogenic and androgenic activities. In this study, we evaluated the interaction of d phenothrin (0, 100, 300 or 1000 mg/kg per day, p.o.) with estrogen- or androgen mediated mechanisms using in vivo short-term assays. While internationally standardized protocols for the uterotrophic and Hershberger assays have not yet been fully developed, both are widely used and are being considered by the OECD as short-term screening assays for hormonal activity. The highest dose level tested for d-phenothrin was a limit dose (1000 mg/kg per day) designated in the current draft protocol by the OECD, and in fact there was no excessive systemic toxicity in both assays; slightly increased liver weight but no change of serum androgen levels in accessing anti-androgenicity. Potential estrogenic effect of d phenothrin was evaluated by means of 3-day uterotrophic assay using immature Crj:CD(SD)IGS rats (20 days of age). No increase in uterine weight (wet or blotted) was observed following oral exposure to d-phenothrin. Reference control ethynyl estradiol (0.001 mg/kg per day) showed a significant effect in this assay protocol. A 10-day Hershberger assay using castrated peripubertal male rats measures the androgenic or anti-androgenic effects of the test chemicals on several accessory glands/tissues (the ventral prostate, dorso-lateral prostate, seminal vesicles with coagulating glands, levator ani plus bulbocavernosus muscles, glans penis and Cowper's glands). d-Phenothrin was administered by oral gavage for 10 days to castrated male Crj:CD(SD)IGS rats (7 weeks of age, rats were castrated at 6 weeks of age) with or without co-administration of 0.2 mg/kg per day testosterone propionate (subcutaneous injection on the dorsal surface). Reference controls of methyltestosterone and p,p'-DDE (100 mg/kg per day) provided significant effects in this assay protocol, whereas d-phenothrin did not show any androgenic or anti-androgenic effects. It is concluded that, based on the results of these two reliable in vivo assays, d-phenothrin exhibits no potential to cause adverse estrogenic or (anti-)androgenic effects even at dose of 1000 mg/kg per day, the limit dose designated in the current draft protocol by the OECD. PMID- 12628316 TI - Automated measurement of acetylcholinesterase activity in rat peripheral tissues. AB - The accepted mechanism of toxicity of many organophosphorous and carbamate insecticides is inhibition of acetylcholinesterase activity. In mammals, part of the toxicity assessment usually includes monitoring blood and/or brain acetylcholinesterase inhibition. Other tissues, however, contain cholinesterase activity (i.e. acetyl- and butyryl-cholinesterase), and the inhibition of that activity may be informative for a full appraisal of the toxicity profile. The present group of studies first optimized the variables for extraction and solubilization of cholinesterase activity from various rat tissues and then refined an existing automated method, in order to differentially assess acetyl and butyrylcholinesterase activity in those tissues. All these studies were conducted using tissues from untreated, Long-Evans, adult rats. The first studies determined the effect of Triton X-100 or salt (NaCl) on the extraction and solubilization of cholinesterase activity from retina, brain, striated muscle, diaphragm, and heart: phosphate buffer plus detergent (1% Triton X-100) yielded the highest activity for most tissues. For striated muscle, however, slightly more activity was extracted if the phosphate buffer contained both 1% Triton X 100 and 0.5 M NaCl. It was also noted that the degree of homogenization of some tissues (e.g. striated muscle) must be increased for maximal solubilization of all cholinesterase activity. Subsequent studies developed a method for assessing the level of acetylcholinesterase, butyrylcholinesterase and total cholinesterase activity in these tissues using an automated analyzer. In conclusion, automated assay of acetylcholinesterase activity in cholinergically innervated tissues in the rat (other than brain) is achievable and relatively convenient. PMID- 12628317 TI - Cadmium exposure and trace elements in human breast milk. AB - The interrelations of the seven elements, calcium (Ca), magnesium (Mg), sodium (Na), potassium (K), phosphorus (P), copper (Cu), zinc (Zn), and cadmium (Cd) in human breast milk were examined in Japanese mothers to clarify the effects of Cd exposure on these important elements for infant growth. Breast milk and urine samples were obtained from 68 mothers, aged 19-38 years, at 5-8 days postpartum. The concentrations were determined by inductively-coupled plasma atomic emission spectrometry for Ca, Mg, Na, K, P, by flame atomic absorption spectrophotometry for Cu and Zn, and by flameless atomic absorption spectrophotometry for Cd. Geometrical mean Cd concentrations were 0.28 (geometrical standard deviation=1.82) microg/l in breast milk and 1.00 (1.93) microg/g creatinine in urine. Among the above elements only Cd concentration in breast milk was significantly correlated with urinary Cd concentration (r=0.451, P<0.001). Significant positive correlations were found between Cu and Ca (r=0.500, P<0.001), Cu and Mg (r=0.378, P<0.01), and Zn and Mg (r=0.355, P<0.01) in breast milk. Cd concentration in breast milk showed an inverse relationship with Ca concentration in breast milk (r=-0.248, P<0.05). These results indicate that the Cd concentration in breast milk closely reflects Cd body burden, with increased Cd in breast milk possibly affecting Ca secretion in breast milk. PMID- 12628319 TI - Microfabricated drug delivery systems: from particles to pores. AB - Microfabrication techniques which permit the creation of therapeutic delivery systems that possess a combination of structural, mechanical, and perhaps electronic features may surmount challenges associated with conventional delivery of therapy. In this review, delivery concepts are presented which capitalize on the strengths of microfabrication. Possible applications include micromachined silicon membranes to create implantable biocapsules for the immunoisolation of pancreatic islet cells-as a possible treatment for diabetes-and sustained release of injectable drugs needed over long time periods. Asymmetrical, drug-loaded microfabricated particles with specific ligands linked to the surface are proposed for improving oral bioavailability of peptide (and perhaps protein) drugs. In addition, microfabricated drug delivery systems ranging from transdermal microneedles to implantable microchips will be discussed. PMID- 12628321 TI - Lab-on-a-chip for drug development. AB - Significant advances have been made in the development of micro-scale technologies for biomedical and drug discovery applications. The first generation of microfluidics-based analytical devices have been designed and are already functional. Microfluidic devices offer unique advantages in sample handling, reagent mixing, separation, and detection. We introduce and review microfluidic concepts, microconstruction techniques, and methods such as flow-injection analysis, electrokinesis, and cell manipulation. Advances in micro-device technology for proteomics, sample preconditioning, immunoassays, electrospray ionization mass spectrometry, and polymerase chain reaction are also reviewed. PMID- 12628320 TI - Biodegradable nanoparticles for drug and gene delivery to cells and tissue. AB - Biodegradable nanoparticles formulated from poly (D,L-lactide-co-glycolide) (PLGA) have been extensively investigated for sustained and targeted/localized delivery of different agents including plasmid DNA, proteins and peptides and low molecular weight compounds. Research about the mechanism of intracellular uptake of nanoparticles, their trafficking and sorting into different intracellular compartments, and the mechanism of enhanced therapeutic efficacy of nanoparticle encapsulated agent at cellular level is more recent and is the primary focus of the review. Recent studies in our laboratory demonstrated rapid escape of PLGA nanoparticles from the endo-lysosomal compartment into cytosol following their uptake. Based on the above mechanism, various potential applications of nanoparticles for delivery of therapeutic agents to the cells and tissue are discussed. PMID- 12628322 TI - Microchip-based chemical and biochemical analysis systems. AB - This review focuses on chemical and biochemical analysis systems using pressure driven microfluidic devices or microchips. Liquid microspace in a microchip has several characteristic features, for example, short diffusion distances, high specific interfacial area and small heat capacity. These characteristics are the key to controlling micro unit operations and constructing new integrated chemical systems. By combining multiphase laminar flow and the micro unit operations, such as mixing, reaction, extraction and separation, continuous flow chemical processing systems are realized in the microchip format. By applying these concepts, several different analysis systems were successfully integrated on a microchip. In this paper, we introduce the microchip-based chemical systems for wet analysis of cobalt ion, multi-ion sensors, immunoassay, and cellular analysis. PMID- 12628323 TI - Micro- and nanobiotechnology for biosensing cellular responses. AB - Cells represent the minimum functional and integrating communicable unit of living systems. Cultured cells both transduce and transmit a variety of chemical and physical signals, i.e., production of specific substances and proteins, throughout their life cycle within specific tissues and organs. Such cellular responses might be usefully employed as parameters to obtain chemical information for both pharmaceutical and chemical safety, and drug efficacy profiles in vitro as a screening tool. However, such cellular signals are very weak and not easily detected with conventional analytical methods. By using micro- and nanobiotechnology methods integrated on-chip, a higher sensitivity and signal amplification has been developed for cellular biosensing. Micro- and nanotechnology is rapidly evolving to open new combinations of methods with improved technical performance, helping to resolve challenging bioanalytical problems including sensitivity, signal resolution and specificity by interfacing these technologies in small volumes in order to confirm specific cellular signals. Integration of cell signals in both rapid time and small space, and importantly, between different cell populations (communication and systems modeling) will permit many more valuable measurements of the dynamic aspects of cell responses to various chosen stimuli and their feedback. This represents the future for cell-based biosensing. PMID- 12628324 TI - PEGylated nanoparticles for biological and pharmaceutical applications. AB - The utility of polymeric micelles formed through the multimolecular assembly of block copolymer was comprehensively described as novel core-shell typed colloidal carriers for drug and gene targeting. Particularly, novel approaches for the formation of functionalized poly(ethylene glycol) (PEG) layers as hydrophilic outer shell were focused to attain receptor-mediated drug and gene delivery through PEG-conjugated ligands with a minimal non-specific interaction with other proteins. Surface organization of block copolymer micelles with cross-linking core was also described from a standpoint of the preparation of a new functional surface-coating with a unique macromolecular architecture. The micelle-attached surface and the thin hydrogel layer made by layered micelles exhibited nonfouling properties and worked as the reservoir for hydrophobic reagents. Furthermore, the potential utility of multimolecular assembly derived from heterobifunctional PEGs and block copolymers were explored to systematically modify the properties of metal and semiconductor nanostructures by controlling their structure and their surface properties, making them extremely attractive for use in biological and biomedical applications. PMID- 12628325 TI - Design strategies to improve soluble macromolecular delivery constructs. AB - Macromolecular therapeutics provide numerous benefits for the delivery of cytotoxic or poorly soluble drugs in vivo. However, these constructs often encounter barriers for drug delivery on both the systemic and subcellular level. Many soluble polymer carriers have been designed to surmount specific physiological barriers individually, but less work has been dedicated to designing an all-encompassing construct that addresses multiple therapeutic barriers at once. Incorporation of multiple agents already individually known to increase effectiveness into one carrier could further improve current drug delivery technology. Recent developments in subcellular delivery of therapeutic agents in soluble macromolecular carriers are discussed in the context of the future possibility for the design of an all-encompassing soluble multi-functional drug delivery vehicle. PMID- 12628327 TI - A trial on regeneration therapy of rat liver cirrhosis by controlled release of hepatocyte growth factor. AB - This paper is an investigation of therapeutic trial on the liver cirrhosis by the controlled release of hepatocyte growth factor (HGF). Biodegradable microspheres were prepared from gelatin for the controlled release of HGF. Rats with liver cirrhosis were prepared by the intraperitoneal injection of thioacetamide at a dose of 0.2 g/kg every other day for 10 weeks. The rats received single intraperitoneal injection of gelatin microspheres incorporating 2 or 0.4 mg of HGF, 2 or 0.4 mg of free HGF, and HGF-free, empty gelatin microspheres 3 weeks after the last thioacetamide injection. Histological observation of the rat liver revealed that injection of gelatin microspheres incorporating HGF effectively allowed to recovery from the liver fibrosis, inducing liver regeneration. When the histological score, the area of fibrous linkage, and the hydroxyproline content in the liver were evaluated, all values were significantly smaller than those of the free HGF and empty gelatin microspheres injection, irrespective of the HGF dose. It is concluded that the gelatin microspheres incorporating HGF are a promising therapeutic method of inducing successful liver regeneration by fibrosis digestion in rats with cirrhosis. PMID- 12628328 TI - POE/PLGA composite microspheres: formation and in vitro behavior of double walled microspheres. AB - The poly(ortho ester) (POE) and poly(D,L-lactide-co-glycolide) 50:50 (PLGA) composite microspheres were fabricated by a water-in-oil-in-water (w/o/w) double emulsion process. The morphology of the composite microspheres varied depending on POE content. When the POE content was 50, 60 or 70% in weight, the double walled microspheres with a dense core of POE and a porous shell of PLGA were formed. The formation of the double walled POE/PLGA microspheres was analysed. Their in vitro degradation behavior was characterized by scanning electron microscopy, gel permeation chromatography, Fourier-transform infrared microscopy and nuclear magnetic resonance spectroscopy (NMR). It was found that compared to the neat POE or PLGA microspheres, distinct degradation mechanism was achieved in the double walled POE/PLGA microspheres system. The degradation of the POE core was accelerated due to the acidic microenvironment produced by the hydrolysis of the outer PLGA layer. The formation of hollow microspheres became pronounced after the first week in vitro. 1H NMR spectra showed that the POE core was completely degraded after 4 weeks. On the other hand, the outer PLGA layer experienced slightly retarded degradation after the POE core disappeared. PLGA in the double walled microspheres kept more than 32% of its initial molecular weight over a period of 7 weeks. PMID- 12628329 TI - Bioadhesive poly(methyl methacrylate) microdevices for controlled drug delivery. AB - Oral delivery is the preferred route of drug administration. However, the breakdown of molecules and low levels of absorption in the gastrointestinal system render the oral delivery of proteins and peptides ineffective. Bioadhesive delivery devices can be used to circumvent these problems by protecting the drug from gastrointestinal denaturation, localizing and prolonging a drug at a specific target site, and maintaining direct contact with the intestinal cells, thereby increasing the drug concentration gradient. Microfabrication technology may offer some potential advantages over conventional delivery technologies. The benefits of microfabrication include the ability to tailor the size, shape, reservoir volume, and surface characteristics of the drug delivery vehicle. In this study, bioadhesive properties were introduced to microfabricated poly(methyl methacrylate) (PMMA) microdevices by attachment of lectins, a group of proteins capable of specifically targeting cells in the gastrointestinal tract. In this process, the PMMA microdevices were chemically modified by aminolysis to yield amine-terminated surfaces. Avidin molecules were covalently bound to the surface of the particles using a hydroxysuccinimide catalyzed carbodiimide reagent and then incubated in an aqueous solution of biotinylated lectin. The lectin-modified microdevices were examined in vitro in terms of their bioadhesive characteristics. PMID- 12628330 TI - Sustained release of human growth hormone from microcapsules prepared by a solvent evaporation technique. AB - Biodegradable microcapsules for sustained release of recombinant human growth hormone (rhGH) were prepared by a solid-in-oil-in-water (S/O/W) emulsion solvent evaporation technique using lyophilized protein microparticles. The minimum mean particle size of rhGH in S/O dispersions was 2.8-3.0 microm when ammonium acetate was added at molar ratios of 10-20 times against rhGH. High entrapment of rhGH in microcapsules was achieved by incorporating rhGH powder with a smaller particle size obtained by lyophilizing with ammonium acetate. As the particle size of rhGH decreased, the in vivo initial release decreased, while subsequent serum levels of rhGH in sustained release phase were higher. Addition of zinc oxide to microcapsules resulted in higher serum levels than those prepared without zinc oxide, suggesting a stabilizing effect of zinc oxide after subcutaneous injection into rats. The release profile of rhGH from microcapsules was controllable by selecting the proper copoly(DL-lactic/glycolic)acid (PLGA) with L/G ratio and molecular weight. Utilization of rhGH powder with a smaller particle size obtained by lyophilizing with ammonium acetate is essential for preparation of microcapsules with high entrapment and well-controlled sustained release profile with small initial release. PMID- 12628331 TI - Novel transdermal drug penetration enhancer: synthesis and enhancing effect of alkyldisiloxane compounds containing glucopyranosyl group. AB - The syntheses of alkyldisiloxanes containing sugar moiety with various alkyl chain length were investigated, in order to develop a silicone-based transdermal penetration enhancer which was expected to show a low irritation to the skin. 1 Alkyl-3-beta-D-glucopyranosyl-1,1,3,3-tetramethyldisiloxanes (Glc-SiCs) were prepared by two-step hydrosilylations of 1-alkene and 1-allyl-beta-D-glucose tetraacetate with 1,1,3,3-tetramethyldisiloxane in the presence of bis(benzonitrile)platinum dichloride as the catalyst, followed by hydrolysis of the acetyl groups with sodium methoxide. The enhancing effect of Glc-SiCs on the percutaneous drug penetration was evaluated by in vitro experiments using a two chamber diffusion cell. Antipyrine (ANP) and indomethacin (IND) were used as hydrophilic and hydrophobic model drugs, respectively, and the amount of drug permeating through the rat abdominal skin with or without Glc-SiCs was estimated by HPLC. As a result, Glc-SiCs exhibited a enhancing effect on the permeation of both drugs through the skin, which was influenced by the alkyl chain length of Glc-SiCs. In addition, it was suggested that a suitable balance of polarity would be necessary to appear the high enhancing effect, where Glc-SiCs with octyl and decyl groups exhibited the highest enhancing effect. From the determination of kinetic parameters in the drug permeation, it was also found that this enhancing effect was due to the increase of both partition and diffusion coefficients of drug permeation through the skin. By experiments to determine the amount of cholesterol extracted from the skin, the defatting effect would be one of the functions of Glc-SiCs which resulted in the high enhancing activity. Furthermore, according to the Draize test, it was confirmed that Glc-SiCs showed a low irritation to the skin. PMID- 12628332 TI - Transdermal delivery of timolol by electroporation through human skin. AB - The purpose was to achieve therapeutic fluxes of timolol by transdermal delivery using skin electroporation. The transdermal transport of timolol through human stratum corneum was studied in three compartment diffusion cells. The electrodes, buffer composition and pulse conditions were optimized. Timolol maleate concentration in the donor compartment was 40 mg/ml. Square wave pulses were applied. Electroporation enhanced the transdermal transport of timolol by 1-2 orders of magnitude as compared to passive diffusion. Even though the current application lasted for only 10 s, the transdermal transport remained high after pulsing for at least 6 h. Higher fluxes were obtained with Pt electrodes close to the skin and a phosphate buffer. 10 pulses of 400 V-10 ms were more efficient than 10 low voltage-long duration pulses. Therapeutic fluxes of timolol (>50 microg/cm(2) per h) through human stratum corneum were achieved by electroporation. PMID- 12628333 TI - Controlled release of verapamil hydrochloride from waxy microparticles prepared by spray congealing. AB - In this work, the potential of waxes for preparing with the ultrasonic spray congealing technique microparticles for controlling the in vitro release of verapamil HCl was investigated. The first part of the study encompassed the optimisation of the formulation to achieve an efficient drug incorporation together with a satisfactory in vitro drug release rate. In particular, microcrystalline wax, stearyl alcohol and mixtures of the two were used. Also a surfactant (soya lecithin) was added to the formulations. After the particle size analysis, the characterisation of the microparticles involved the study of the solid state of drug and carriers in the systems (DSC, HSM and XRD) and the morphological and chemical analyses of the microparticle surface (SEM and XPS). Finally, the drug release mechanism from these devices was evaluated using the statistical moment analysis. The results of this study show that by selecting the type and the amount of the carriers, microparticles with a spherical shape and a good encapsulation efficiency were observed. These particles showed a zero-order release for 8 h, without modifying the solid state properties of the drug. Therefore, waxy microparticles prepared by the ultrasonic spray congealing technique are promising solvent-free devices for controlling the release of verapamil HCl. PMID- 12628334 TI - Bioadhesive film for the transdermal delivery of lidocaine: in vitro and in vivo behavior. AB - The aim of this work was to study, in vitro and in vivo, the behavior of a skin bioadhesive film containing lidocaine. The film characterization included drug transport studies through skin in vitro and in vivo tape stripping with and without iontophoresis. We studied the effect of drug loading in order to identify the release mechanism. Finally, the release rate was compared with a lidocaine commercial gel, to assess the therapeutic value. From the data obtained it can be concluded that the monolayer film acts as a water-permeable transdermal/dermal patch on application to the skin. The permeation kinetics across the skin was not linear, but the patch acted as a matrix controlling drug delivery. Additionally, the permeation rate increased with drug loading. The in vivo experiments with tape stripping indicated that the presence of water during film application is essential to achieve not only the proper adhesion but also an effective accumulation. The application of electric current to the patch can further increase the amount of drug accumulated in the stratum corneum. PMID- 12628335 TI - Transdermal iontophoresis of insulin. V. Effect of terpenes. AB - To increase the skin permeation of large peptides like insulin, it is necessary to utilize a combination of enhancement strategies. In this regard, this study investigated the effect of terpenes/EtOH combination in comparison to EtOH and neat terpene on transdermal iontophoretic permeation of insulin. Ex-vivo experiments were conducted using full thickness rat skin after pre-treatment for 2 h with 5% of menthol, menthone, cineole and pulegone in EtOH; EtOH alone; neat menthone with and without iontophoresis (0.5 mA/cm(2); 6 h). FT-IR studies were carried out using rat epidermal sheets after pre-treatment with enhancer solution for 2 h and tritiated water permeation studies was used to investigate the alteration in skin barrier property after enhancer or current treatment. The lag time was significantly reduced (P<0.05) with terpene/EtOH pre-treatment in comparison to passive control and EtOH pre-treatment, although there was no significant difference (P>0.05) among the terpenes. Synergistic enhancement in flux was observed with terpene/EtOH, and menthone/EtOH showed highest enhancement among the terpene/EtOH combinations. On the other hand, enhancement with neat menthone was higher than with menthone/EtOH. FT-IR studies showed that terpene/EtOH, EtOH and neat terpene act at the intercellular lipids. The skin barrier property was significantly (P<0.05) compromised with neat menthone treatment. Iontophoresis had a lesser effect on skin barrier property compared to chemical enhancer pre-treatment. Terpene/EtOH caused synergistic enhancement of insulin permeation when combined with iontophoresis and was influenced by the type and concentration of terpene. PMID- 12628336 TI - Tumor targeting of gene expression through metal-coordinated conjugation with dextran. AB - Tumor targeting of plasmid DNA was achieved through the conjugation of dextran derivatives with chelate residues based on metal coordination. Diethylenetriamine pentaacetic acid (DTPA), spermidine (Sd), and spermine (Sm) were chemically introduced to the hydroxyl groups of dextran to obtain dextran-DTPA, dextran-Sd and dextran-Sm derivatives. Conjugation of the dextran derivative by Zn(2+) coordination decreased the apparent size of the plasmid DNA, depending on the derivative type. The negative zeta potential of plasmid DNA became almost 0 mV after Zn(2+)-coordinated conjugation with dextran-Sm. When the dextran derivative plasmid DNA conjugates with Zn(2+) coordination were intravenously injected subcutaneously into mice bearing Meth-AR-1 fibrosarcoma, the dextran-Sm-plasmid DNA conjugate significantly enhanced the level of gene expression in the tumor, in contrast to the conjugate of other dextran derivatives and free plasmid DNA. The enhanced gene expression produced by the Zn(2+)-coordinated dextran-Sm plasmid DNA conjugate was specific to the tumor, whereas a simple mixture of dextran-Sm and plasmid DNA was not effective. The level of gene expression depended on the percentage of chelate residues introduced, the mixing weight ratio of the plasmid DNA/Sm residue used for conjugate preparation, and the plasmid DNA dose. A fluorescent microscopic study revealed that localization of plasmid DNA in the tumor tissue was observed only after injection of the dextran Sm-plasmid DNA conjugate with Zn(2+) coordination. In addition, the gene expression induced by the conjugate lasted for more than 10 days after the injection. We conclude that Zn(2+)-coordinated dextran-Sm conjugation is a promising way to enable plasmid DNA to target the tumor in gene expression as well as to prolong the duration of gene expression. PMID- 12628337 TI - Remarkable induction of apoptosis in cancer cells by a novel cationic liposome complexed with a bcl-2 antisense oligonucleotide. AB - We reported recently a novel cationic cholesterol derivative with a hydroxyethylamino head group, cholesteryl-3beta-carboxyamidoethylene-N hydroxyethylamine (I) for liposome-mediated gene transfection [FEBS Lett., 408 (1997) 232]. In the present paper we have studied whether this novel cationic liposome is prominent in nature to suppress cell growth of human cancer cells. Bcl-2 antisense phosphorothioate oligonucleotides (AS-ODNs) were complexed with the cationic liposomes with the derivative (I) and they were introduced into human cervix epithelial carcinoma cell lines HeLa, and mouse fibroblast NIH3T3 cells. An AS-ODNs targeting/bcl-2 gene induced probably apoptosis (including necrosis in some cases) in HeLa and NIH3T3 cells, however, nonsense oligonucleotides (NS-ODNs) corresponding to a scrambled-sequence control hardly induced apoptosis. Induction of apoptosis was much greater than that by commercially available DC-Chol liposomes. Fluorescence intensities of FITC conjugated bcl-2 AS-ODNs were specifically found in the nucleus. The intensity of the AS-ODNs was mostly consistent with the amounts of Bcl-2 proteins observed by Western blot analysis in the target cells. The results showed the possibility that this new cationic cholesterol derivative might be very promising to be used for liposome-mediated gene targeting in vitro and in vivo. PMID- 12628338 TI - Sustained release of plasmid DNA using lipid microtubules and agarose hydrogel. AB - Non-viral gene therapy typically results in low transfection efficiencies and transient gene expression. To address these limitations, two sustained delivery systems capable of releasing functional, compacted DNA for over 50 days were designed. A luciferase plasmid was compacted with a polylysine-polyethylene glycol conjugate and released from agarose hydrogel and lipid microtubule hydrogel delivery systems for over 50 days. The released DNA was characterized structurally using sedimentation, electron microscopy, and serum stability, and functionally using in vitro transfections. The released DNA retained its physical compaction and nuclease resistance and was converted from supercoiled to nicked and linear forms. Released compacted DNA produced significant gene expression in vitro, although at lower levels than freshly compacted DNA. Thus, hydrogels and lipid microtubules successfully provided the slow release of bioactive, compacted DNA. PMID- 12628339 TI - Prevention of autoimmune insulitis by delivery of a chimeric plasmid encoding interleukin-4 and interleukin-10. AB - The combined administration of interleukin-4 (IL-4) and interleukin-10 (IL-10) expression plasmids has demonstrated synergistic effects on the prevention of autoimmune diabetes. To this end, we constructed a co-expression 'chimeric' plasmid, pCMV-IL4-IL10, in which the expression of IL-4 and IL-10 was driven by two separate CMV immediate early promoters by using the biodegradable polymer, poly[alpha-(4-aminobutyl)-L-glycolic acid] (PAGA) as a gene carrier to optimize gene delivery. In vitro transfection assays of the chimeric plasmid in 293T cells showed higher expression levels as well as dose dependence than the single gene expression plasmids. To evaluate the in vivo efficacy of the chimeric plasmid, the pCMV-IL4-IL10/PAGA complex was intravenously injected into 4-week-old non obese diabetic (NOD) mice and compared to the co-administration group. While both groups had persistent gene expression longer than 5 weeks, the IL-4 and IL-10 serum levels of the chimeric group were higher than those in the co administration group. Furthermore, the degree of insulitis in the chimeric group was improved over both the co-administration and non-injected control groups. These results suggest that the chimeric IL-4 and IL-10 expression plasmid can effectively reduce the incidence of autoimmune insulitis. PMID- 12628341 TI - The next ice age: cryo-electron tomography of intact cells. AB - Recent advances in electron tomography are beginning to reveal the internal structure of eukaryotic cells in their native states in three dimensions at molecular resolution. These observations represent the culmination of years of effort to develop protocols for automated data collection, image reconstruction and cryogenic preservation. Cryo-tomograms of Dictyostelium cells depict distinct populations of ribosomes, proteasomes and networks of actin filaments interconnected by branching or bundling, apparently controlled by strategically placed actin-associated proteins. PMID- 12628342 TI - Down to the origin: Cdc6 protein and the competence to replicate. AB - In eukaryotes, DNA replication requires the regulated assembly of pre-replicative complexes (pre-RCs) onto DNA during G1 phase. Pre-RCs render the chromatin competent to replicate, yet it is only at the G1-S phase transition that protein kinase complexes trigger the transition to DNA replication. Central to the formation of pre-RCs and regulation of DNA replication is the Cdc6 protein. Two recent studies have shown that Cdc6 is the long-sought factor that confers the competence to replicate in unfertilized Xenopus eggs. PMID- 12628343 TI - Rotary protein motors. AB - Three protein motors have been unambiguously identified as rotary engines: the bacterial flagellar motor and the two motors that constitute ATP synthase (F(0)F(1) ATPase). Of these, the bacterial flagellar motor and F(0) motors derive their energy from a transmembrane ion-motive force, whereas the F(1) motor is driven by ATP hydrolysis. Here, we review the current understanding of how these protein motors convert their energy supply into a rotary torque. PMID- 12628344 TI - The "Gab" in signal transduction. AB - Tyrosine phosphorylation plays an important role in controlling cellular growth, differentiation and function. Abnormal regulation of tyrosine phosphorylation can result in human diseases such as cancer. A major challenge of signal transduction research is to determine how the initial activation of protein-tyrosine kinases (PTKs) by extracellular stimuli triggers multiple downstream signaling cascades, which ultimately elicit diverse cellular responses. Recent studies reveal that members of the Gab/Dos subfamily of scaffolding adaptor proteins (hereafter, "Gab proteins") play a crucial role in transmitting key signals that control cell growth, differentiation and function from multiple receptors. Here, we review the structure, mechanism of action and function of these interesting molecules in normal biology and disease. PMID- 12628345 TI - Making vascular networks in the adult: branching morphogenesis without a roadmap. AB - The vascular system is unique in that extensive branching morphogenesis may take place in the adult. Developmental neovascularization is guided by precise spatial cues but vessel formation in the adult is not genetically programmed. Here, we review different adult modes for branch patterning, acquiring artery or vein identity and allocating vascular progenitor cells. The endothelium shows a remarkable degree of self-organization into a treelike network and hemodynamic forces are important in rectifying abnormal branching. This discussion is in the context of a contemplated therapy for improving organ perfusion by creating new vascular loops properly integrated within the existing network. PMID- 12628346 TI - At the acidic edge: emerging functions for lysosomal membrane proteins. AB - It has recently become clear that lysosomes have more complex functions than simply being the end-point on a degradative pathway. Similarly, it is now emerging that there are interesting functions for the limiting membranes around these organelles and their associated proteins. Although it has been known for several decades that the lysosomal membrane contains several highly N glycosylated proteins, including the lysosome-associated membrane proteins LAMP-1 and LAMP-2 and lysosomal integral membrane protein-2/lysosomal membrane glycoprotein-85 (LIMP-2/LGP85), specific functions of these proteins have only recently begun to be recognized. Although the normal functions of LAMP-1 can be substituted by the structurally related LAMP-2, LAMP-2 itself has more specific tasks. Knockout of LAMP-2 in mice has revealed roles for LAMP-2 in lysosomal enzyme targeting, autophagy and lysosomal biogenesis. LAMP-2 deficiency in humans leads to Danon disease, a fatal cardiomyopathy and myopathy. Furthermore, there is evidence that LAMP-2 functions in chaperone-mediated autophagy. LIMP-2/LGP85 also seems to have specific functions in maintaining endosomal transport and lysosomal biogenesis. The pivotal function of lysosomal membrane proteins is also highlighted by the recent identification of disease-causing mutations in cystine and sialic acid transporter proteins, leading to nephropathic cystinosis and Salla disease. PMID- 12628347 TI - Transcriptional repression by Myc. AB - The Myc oncoprotein is a transcription factor that can both activate and repress genes. Transcriptional activation by Myc is well understood, but, by contrast, the mechanisms through which Myc represses transcription have remained elusive. Recent evidence suggests that complex formation by Myc with a zinc-finger transcription factor, Miz-1, plays an important role in mediating repression by Myc. The findings might explain how Myc interferes with cell-cycle arrest in response to TGF-beta, APC and DNA damage. PMID- 12628348 TI - An array of insights: application of DNA chip technology in the study of cell biology. AB - The advent of DNA microarray technology has ushered in an era of systems biology whereby researchers can study the transcriptional behavior of thousands of genes in parallel. Advances in manufacturing techniques and informatics, and the availability of several genome sequences have furthered these capabilities to the point where whole-transcriptome studies can be accomplished in yeast, flies and plants, and soon will be possible in mammals. Concomitant with the expanding ability of the technology has been the development of novel techniques and their application towards the study of cellular biology. PMID- 12628349 TI - Is there a physiological role for the neurosteroid THDOC in stress-sensitive conditions? AB - Endogenous neurosteroids affect brain excitability during physiological states such as pregnancy and the menstrual cycle, and during conditions of acute and chronic stress. The neurosteroid allotetrahydrodeoxycorticosterone (THDOC) is an allosteric modulator of the GABA(A) receptor. Although the role of THDOC within the brain is undefined, recent studies indicate that stress induces THDOC to levels that can activate GABA(A) receptors. These results might have significant implications for human stress-sensitive conditions such as epilepsy, post traumatic stress disorder and depression. PMID- 12628350 TI - Salvinorin A: the "magic mint" hallucinogen finds a molecular target in the kappa opioid receptor. AB - Salvinorin A, a neoclerodane diterpene, is the most potent naturally occurring hallucinogen known and rivals the synthetic hallucinogen lysergic acid diethylamide in potency. Recently, the molecular target of salvinorin A was identified as the kappa opioid receptor (KOR). Salvinorin A represents the only known non-nitrogenous KOR selective agonist. Based on the selectivity of salvinorin A for the KOR, this receptor represents a potential molecular target for the development of drugs to treat disorders characterized by alterations in perception, including schizophrenia, Alzheimer's disease and bipolar disorder. PMID- 12628351 TI - GIP or not GIP? That is the question. AB - Glucose-dependent insulinotropic polypeptide (GIP) is a gastrointestinal hormone that is released during a meal and facilitates the disposal of both glucose and fat. Because of the insulinotropic action of GIP, this hormone has been considered as a potential therapy of type 2 diabetes, where insulin secretion is inadequate. However, a recent study using GIP receptor knockout mice suggests that inhibition of GIP signalling might be a new target for anti-obesity drugs. PMID- 12628352 TI - Statins and myotoxicity. PMID- 12628353 TI - Pitfalls in estimating drug-related crash risk. PMID- 12628355 TI - A common thread for pain and memory synapses? Brain-derived neurotrophic factor and trkB receptors. AB - Recent evidence indicates that trophic factors can exert fast effects on neurones and so alter synaptic plasticity. Here, we focus on brain-derived neurotrophic factor (BDNF), which exerts a modulatory action at hippocampal synapses that are involved in learning and memory, and at the first pain synapse between primary sensory neurones and dorsal horn neurones. Hippocampal and sensory neurones share some properties for the release of endogenous BDNF. In the Schaffer collateral pathway of the hippocampus, binding of BDNF to high-affinity trkB receptors is essential for the induction of long-term potentiation, a specific type of synaptic plasticity. However, the consequences of BDNF binding to trkB receptors in the dorsal horn in relation to pain mechanisms are less well established and are considered in detail. PMID- 12628356 TI - Education: Teaching pharmacogenomics to prepare future physicians and researchers for personalized medicine. AB - The vision of personalized medicine, the practice of medicine where each patient receives the most appropriate medical treatments and the most fitting dosage and combination of drugs based on his or her genetic make-up, seems to become more realistic as our knowledge about the human genome rapidly expands. We already know the reason for many types of adverse drug reactions, which are often related to polymorphic gene alleles of drug metabolizing enzymes. Moreover, insight into reasons for poor drug efficacy, often related to single nucleotide polymorphisms or larger polymorphisms in genes encoding drug target proteins, has been gained. There is a growing need to incorporate this increasingly complex body of knowledge to the standard curriculum of medical schools, so that the forthcoming generation of clinicians and researchers will be familiar with the latest developments in pharmacogenomics and medical bioinformatics, and will be capable of providing patients with the expected benefits of personalized medicine. PMID- 12628357 TI - Viral leads for chemokine-modulatory drugs. AB - The chemokine system, which controls leukocyte trafficking, provides several potentially very attractive anti-inflammatory drug targets. However, the complexity and redundancy of this system makes it very difficult to exploit through classical drug discovery. Despite this, viruses have millions of years of experience in manipulating this system. For example, virally encoded "biopharmaceuticals"--chemokines and chemokine binding proteins--demonstrate the effectiveness of blocking a carefully selected group of chemokine receptors and how the local immune response can be changed from one dominated by Th1 cells to one dominated by Th2 cells by targeting specific chemokine receptors. The crucial importance of the binding of chemokines to glycosaminoglycans to produce their effects is also highlighted by viruses that produce binding proteins to disrupt the gradient of chemokines, which guides the direction leukocyte migration. PMID- 12628358 TI - Immunomodulation in multiple sclerosis: from immunosuppression to neuroprotection. AB - Multiple sclerosis (MS) is the most common disabling neurological disease of young adulthood. Following advances in the understanding of the immunological mechanisms that underlie the pathogenesis of MS, a growing arsenal of immunomodulatory agents is available. Two classes of immunomodulators are approved for long-term treatment of MS, the efficacy of several promising new concepts is being tested in clinical trials and classical immunosuppressive agents used in MS treatment have been shown to exert specific, immunomodulatory effects. Furthermore, two recent observations have changed our basic understanding of the pathogenesis of MS. First, immune cells in MS lesions have neuroprotective activity, which indicates a beneficial role of neuroinflammation. Second, there is evidence that axonal loss, rather than demyelination, underlies the progression of MS and, hence, constitutes a therapeutic target. PMID- 12628360 TI - Techniques: Visualizing apoptosis using nuclear magnetic resonance. AB - Apoptosis plays a key role in tumour biology, and the induction of apoptosis forms a cornerstone of most anticancer therapies. New developments in nuclear magnetic resonance spectroscopy (MRS) and magnetic resonance imaging (MRI) have taken these techniques far beyond their original roles as the workhorses of structural and pharmaceutical chemistry and clinical imaging to the detection of previously inaccessible and unrecognized biological phenomena in living cells and tissues undergoing apoptosis. These new MR techniques can be used in the development of new drugs and in the improved detection of treatment responses in the clinic. PMID- 12628359 TI - Cell-cycle dysregulation and anticancer therapy. AB - Cell-cycle dysregulation is a hallmark of tumor cells. The ability of normal cells to undergo cell-cycle arrest after damage to DNA is crucial for the maintenance of genomic integrity. The biochemical pathways that stop the cell cycle in response to cellular stressors are called checkpoints. Defective checkpoint function results in genetic modifications that contribute to tumorigenesis. The regulation of checkpoint signaling also has important clinical implications because the abrogation of checkpoint function can alter the sensitivity of tumor cells to chemotherapeutics. Here, we provide an overview of the mechanisms that regulate the cell cycle, current anticancer therapies that target checkpoint signaling pathways, and strategies for the development of novel chemotherapeutic agents. PMID- 12628362 TI - Molecular computing revisited: a Moore's Law? AB - Moore's Law states that the processing power of microchips doubles every one to two years. This observation might apply to the nascent field of molecular computing, in which biomolecules carry out logical operations. Incorporation of new technologies that improve sensitivity and throughput has increased the complexity of problems that can be addressed. It is an ultimate goal for molecular computers to use the full potential of massive parallelism. PMID- 12628361 TI - Improving plant genetic engineering by manipulating the host. AB - Agrobacterium-mediated transformation is a major technique for the genetic engineering of plants. However, there are many economically important crop and tree species that remain highly recalcitrant to Agrobacterium infection. Although attempts have been made to "improve" transformation by altering the bacterium, future successes might come from manipulation of the plant. Recent studies that identified several plant genes involved in Agrobacterium-mediated transformation, and their over-expression in currently transformable species, suggest that this approach holds great promise for improving the transformation of recalcitrant, but agronomically important, crops. PMID- 12628363 TI - From human genes to stem cells: new challenges for patent law? AB - The social controversies that have surrounded human cloning and the use of embryos for research purposes might create unique patent issues for stem cell researchers. Policy makers should learn from the legal and ethical concerns associated with human gene patents and develop coherent patent policies that recognize and clearly address emerging social controversies. PMID- 12628364 TI - A new science-business paradigm in anticancer drug development. AB - Studies into the molecular mechanisms of cancer have revealed that, with a few exceptions, the disease lacks a specific drug target. Therefore, new anticancer drugs not only take many years and much money to develop but also might not outperform existing drugs. However, what are assumed to be unfavorable hallmarks of cancer, for example drug resistance, can be exploited for selective anticancer therapy and for protection of normal cells. Based on this paradigm, the drug discovery can be complemented by novel use of existing agents and even "failed" drugs. The pharmacological industry could develop low cost, effective therapeutic modalities, by "re-using" already marketed and late-stage products in cancer selective therapeutic kits. PMID- 12628365 TI - Escherichia coli gets a new virus but it's nothing to sneeze at. AB - For the past two decades virologists have strived to make full-length clones of viral genomes that, on transfection into permissive eukaryotic cells, initiate a productive infection. The large variety of viral RNA and DNA genome structures, as well as different replication strategies, has required investigators to develop new approaches to produce infectious DNA in Escherichia coli. A member of the poxviridae, one of the most complex virus families, has now been made into an infectious clone in E. coli for the first time. Although the isolation was complicated, the infectious clone will greatly simplify future genetic studies of the virus. PMID- 12628366 TI - Controversies over stem cell research. AB - Much interest and effort has focused on the therapeutic potential of stem cell technology to treat presently intractable diseases. However, this scientific promise has been accompanied by important issues, including ethical hurdles, political policies and dilemmas concerning cell-source selection (embryonic versus adult stem cells). Although the contribution of stem cells to medical research seems enormous, many countries now face complex ethical and regulatory questions, which could represent a significant limitation to medical progress and could take years to overcome. PMID- 12628367 TI - Biomedical databases: protecting privacy and promoting research. AB - When combined with medical information, large electronic databases of information that identify individuals provide superlative resources for genetic, epidemiology and other biomedical research. Such research resources increasingly need to balance the protection of privacy and confidentiality with the promotion of research. Models that do not allow the use of such individual-identifying information constrain research; models that involve commercial interests raise concerns about what type of access is acceptable. Researchers, individuals representing the public interest and those developing regulatory guidelines must be involved in an ongoing dialogue to identify practical models. PMID- 12628368 TI - Latest development in viral vectors for gene therapy. AB - Gene therapy includes the application of various viral vectors, which represent most types and families of viruses, suitable for infection of mammalian host cells. Both hereditary diseases and acquired illnesses, such as cancer, can be targeted. Because of the various properties of each viral vector, the definition of their application range depends on factors such as packaging capacity, host range, cell- or tissue-specific targeting, replication competency, genome integration and duration of transgene expression. Recent engineering of modified viral vectors has contributed to improved gene delivery efficacy. PMID- 12628369 TI - Secondary plant metabolites in phytoremediation and biotransformation. AB - For millennia, secondary plant metabolites have antagonized microorganisms, insects and humans alike, ultimately generating a complex and dynamic mixture of facultative and obligate interactions from symbioses to pathogenicity. Secondary plant metabolites have an important role in developing the myriad of organic pollutant-degrading enzymes found in nature. The link between secondary plant metabolites and enzymatic diversity has yet to be exploited, with potential applications in fields as varied as pest management, bioremediation and fine chemical production. PMID- 12628370 TI - Biocatalytic transformations in ionic liquids. AB - Room temperature ionic liquids are non-volatile, thermally stable and highly polar; they are also moderately hydrophilic solvents. Here, we discuss their use as reaction media for biocatalysis. Enzymes of widely diverging types are catalytically active in ionic liquids or aqueous biphasic ionic liquid systems. Lipases, in particular, maintain their activity in anhydrous ionic liquid media; the (enantio)selectivity and operational stability are often better than in traditional media. The unconventional solvent properties of ionic liquids have been exploited in biocatalyst recycling and product recovery schemes that are not feasible with traditional solvent systems. PMID- 12628371 TI - Enzyme activities in fish spermatozoa with focus on lactate dehydrogenase isoenzymes from herring Clupea harengus. AB - The activities of NAD- and NADP-dependent dehydrogenases and creatine kinase were compared in extracts of spermatozoa from herring (Clupea harengus), carp (Cyprinus carpio) and catfish (Clarias gariepinus). The activity of malic enzyme in herring spermatozoa was approximately 5 and 36 times higher than in carp and catfish spermatozoa. In contrast, lactate dehydrogenase activity in herring spermatozoa was very low. Herring spermatozoa possess two isoenzymes of lactate dehydrogenase: LDH-A(2)B(2) and LDH-B(4). Both herring spermatozoa isozymes were separated, partly purified and characterized by kinetic and physico-chemical properties. The pH optima and K(m) values for pyruvate reduction were 7.1, 7.25, 7.6 and 0.22, 0.07, 0.09 mM for LDH-A(4), LDH-A(2)B(2) and LDH-B(4), respectively. The isoenzymes also have different thermostabilities. High activity of malic enzyme in herring spermatozoa suggests adaptation to metabolism at high oxygen tension. PMID- 12628372 TI - Histone-like proteins from Atlantic cod milt: stimulatory effect on Atlantic salmon leucocytes in vivo and in vitro. AB - This study was carried out to reveal some characteristics of cationic proteins from Atlantic cod (Gadus morhua) milt chromatin and to investigate their ability to activate Atlantic salmon (Salmo salar) macrophages. Cationic proteins extracted from cod milt chromatin were fractionated on a cation exchange chromatography column. SDS-PAGE and amino acid analyses of the resulting fractions indicated that these proteins are similar to calf thymus histones. Two cationic protein fractions were used to stimulate leucocytes from Atlantic salmon in vitro and in vivo. Increased production of superoxide, measured as reduction of nitroblue tetrazolium (NBT), was used as indication of macrophage activation. Both fractions induced elevated superoxide anion production in the macrophages after 3 and 6 days of in vitro stimulation. Intraperitoneal injection of the cationic protein fractions in Atlantic salmon (100 mg kg(-1)) four days prior to slaughtering stimulated superoxide production when assayed after one and two days of cell cultivation. In macrophages from fish slaughtered two days after injection, activation could first be seen after two days of cell cultivation. PMID- 12628373 TI - cDNA cloning and characterization of an osmotically sensitive TRP channel from ascidian eggs. AB - The transient receptor potential (TRP) ion channels are thought to be involved in the entry of calcium ion into cells. In this study, we isolated a cDNA clone, HrTRPV, that shows high homology to Caenorhabditis elegans OSM-9, a TRPV subfamily member of the TRP family, from a Halocynthia roretzi fertilized egg cDNA library. We analyzed its properties using HrTRPV-transfected cells. Upon reduction of extracellular osmolarity, the intracellular calcium concentration was found to increase in HrTRPV-transfected cells. This increase in intracellular calcium concentration was dependent on the presence of extracellular calcium ion and was inhibited by treatment with gadolinium ion, a stretch-activated calcium channel blocker. Thus, these results indicate that ascidian egg HrTRPV is an osmotically sensitive TRP channel. PMID- 12628374 TI - The dual activity of pyruvate kinase type M2 from chromatin extracts of neoplastic cells. AB - Pyruvate kinase type M(2) from Morris hepatoma 7777 tumour cell nuclei and cytosol, in contrast to types L and M(2) from nuclei and cytosol of normal rat liver, shows the histone H(1) kinase activity. Moreover, in the presence of L cysteine and without ADP it converts 2-phosphoenolpyruvate (PEP) to pyruvate while in the presence of L-arginine or L-histidine does not. L-Cysteine markedly stimulates the activity of histone H(1) kinase transferring a phosphate group from PEP to, as results suggested, the epsilon -amino group of L-lysine of histone H(1). This, L-cysteine which is known to inhibit the activity of pyruvate kinase type M(2) from neoplastic cells transfering a phosphate from PEP to ADP, can act as a control factor champing the direction of enzymatic reaction in cancer cells. PMID- 12628375 TI - Sterol composition of gonad, muscle and digestive gland of Pecten maximus from Malaga (South Spain). AB - Sterol composition and content and their seasonal variations over 18 months were investigated in adductor muscle, digestive gland and gonads of Pecten maximus. Sterols were isolated by Silicagel 60 thin layer chromatography and identified by gas chromatography/mass spectrometry. Eleven sterols were identified, with cholesterol, brassicasterol, 24-methylenecholesterol and 22-trans dehydrocholesterol being the principal components. The same sterols were found in all three tissues independent of season. The relative amounts of each sterol present in each tissue differed. Total sterol levels in gonad and muscle were higher than in digestive gland. Statistically significant differences (P<0.05) were found between the concentrations of each of the sterols isolated from the gonad or muscle and digestive gland. The seasonal variations in the sterol content of the gonad seem be related to the reproductive cycle, while the sterol content of the digestive gland appears to be linked to diet, mainly diatoms or dinoflagellates. The muscle sterol content showed minor changes throughout the year. PMID- 12628376 TI - Hydrocarbons in the surface wax of eggs and adults of the Colorado potato beetle, Leptinotarsa decemlineata. AB - The major components of the egg hydrocarbons were dimethylalkanes (40%) and trimethylalkanes (24%) in which the first methyl branch was on carbon 2. The major dimethylalkanes were an approximately 2:1 mixture of 2,10- and 2,6 dimethyloctacosanes in females and eggs. The major trimethylalkanes were a mixture of 2,10,16- and 2,10,18-trimethyloctacosanes. 2,x- and 2,x,y-methyl branched alkanes with an odd-numbered carbon backbone were proposed to have an even number of carbon atoms between the first and second methyl branch points indicating that their biosynthesis started with a primer derived from leucine. 13,17,21,25-Tetramethylheptatriacontane was the only tetramethylalkane identified. Females and eggs had more hydrocarbons with a 2-methyl branch point than did the males. The eggs had the lowest amount of internally-branched dimethylalkanes but the largest amount of 2,x-dimethylalkanes in their surface hydrocarbons. Only trace amounts of n-alkanes and alkenes were detected in the surface hydrocarbons of adult males and females, larvae and eggs, of the Colorado potato beetle. PMID- 12628377 TI - Vitellogenin isolation, purification and antigenic cross-reactivity in three teleost species. AB - Vitellogenin (Vtg) was isolated from male greenback flounder (Rhombosolea tapirina), rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) plasma, following induction by estradiol (E(2)) inoculation. The molecular weight of each native molecule, as determined by gel filtration, was 540, 383 and 557 kDa, respectively. With sodium dodecyl sulphate polyacrylamide gel electrophoresis under reducing conditions, Atlantic salmon and greenback flounder Vtg appeared as three major bands (approximately 159, 117, 86 kDa and 155, 104, 79 kDa, respectively), whereas rainbow trout Vtg appeared as one major band (approximately 154 kDa). Several minor bands were also present in each Vtg isolate. Polyclonal antisera, produced against only the highest molecular weight band from each species following excision from reducing gels, were reactive with all major bands in Western blots. In competition ELISA, parallel binding slopes were demonstrated between purified Vtg and plasma from vitellogenic females of the same species, but there was no reaction with plasma from untreated males. These antisera were highly species-specific and little cross-reactivity was noted, even between the two salmonid species. These data suggest that excision of bands from gels is a simple procedure for the preparation of species-specific antisera, and confirm that cross-species assays give highly variable results. PMID- 12628378 TI - Purification and characterization of cathepsin L in arrowtooth flounder (Atheresthes stomias) muscle. AB - A predominant, heat-activated proteinase in muscle extract of arrowtooth flounder (Atheresthes stomias) was purified to 55-fold by heat treatment, followed by a series of chromatographic separations. The apparent molecular mass of the purified enzyme was 27 kDa by size exclusion chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The proteinase had high affinity and activity toward Z-Phe-Arg-NMec with K(m) and k(cat) values of 8.2 microM and 12.2/s, respectively. Activity was inhibited by sulfhydryl reagents and activated by reducing agents. The purified proteinase displayed optimal activity at pH 5.0 5.5 and 60 degrees C, respectively. Consistent with the properties of proteases from other species, the heat-activated proteinase in arrowtooth flounder can be identified as cathepsin L. PMID- 12628379 TI - Cuticular proteins from the horseshoe crab, Limulus polyphemus. AB - Proteins were purified from the carapace cuticle of a juvenile horseshoe crab, Limulus polyphemus, and several of them were characterized by amino acid sequence determination. The proteins are small (7-16 kDa) and their isoelectric points range from 6.5 to 9.2. They have high contents of tyrosine, ranging from 13.5 to 35.4%. Some of the proteins show sequence similarity to cuticular proteins from other arthropod groups, with the most pronounced similarity to proteins from the cuticle of the spider Araneus diadematus. Two proteins show sequence similarity to a hexamerin storage protein from Blaberus discoidalis. PMID- 12628380 TI - Calcium/calmodulin-dependent protein kinase I in Xenopus laevis. AB - Calcium/calmodulin (CaM) dependent protein kinase I (CaM-KI) is a member of a well-defined multi-functional CaM-K family, but its physiological and developmental functions have yet to be determined. Here, we have cloned two cDNAs encoding CaM-KI from a Xenopus laevis (X. laevis) oocyte cDNA library. One is a novel isoform of CaM-KI, named CaM-KI LiKbeta (XCaM-KI LiKbeta). The other is an alpha isoform of CaM-KI (XCaM-KIalpha), which is a highly related to previously cloned mammalian isoform. XCaM-KIalpha was constantly expressed through embryogenesis, whereas XCaM-KI LiKbeta expression dramatically increased in the neurula stage. Both XCaM-KI isoforms exhibited kinase activity in a Ca(2+)/CaM dependent manner. Overexpression of a constitutively active mutant of CaM-KI isoforms inhibited cell cleavage in X. laevis embryos and caused a marked change of cell morphology in Hela cells. Taken together, these results suggest that CaM KI plays a role in cell-structure regulation during early embryonic development. PMID- 12628381 TI - Two novel Bv8-like peptides from skin secretions of the toad Bombina maxima. AB - Two novel bioactive peptides were purified from skin secretions of the toad Bombina maxima. The partial N-terminal sequences of these two peptides were determined by automated Edman degradation. This allowed the cloning of full length cDNAs encoding these two peptides from a cDNA library prepared from the toad skin. The deduced complete amino acid sequences indicate that both peptides are composed of 77 amino acids. A FASTA search in the databanks revealed that they exhibit 86-91% sequence identity with Bv8, a peptide originally isolated from skin secretions of Bombina variegata. They were thus named as Bv8-like peptide 1 (Bv8-LP1) and Bv8-like peptide 2 (Bv8-LP2), respectively. Sequence differences between Bv8-LP1 and 2 were due to six amino acid substitutions at positions 6, 11, 23, 24, 62 and 63. Bv8-LP1 and 2 differed from Bv8 with eleven and seven amino acid substitutions, respectively. Like Bv8, Bv8-LP1 and 2 possessed contractile activity on isolated guinea pig ileum. Additionally, they stimulated contraction of rabbit aortic rings in a dose-dependent manner at nanomolar concentrations. PMID- 12628382 TI - Distribution of rhodanese in tissues of goat (Capra hircus). AB - Rhodanese (thiosulfate: cyanide sulfurtransferase, EC. 2.8.1.1) is a ubiquitous enzyme present in all living organisms, from bacteria to humans and plays a central role in cyanide detoxification. The purpose of this investigation is to determine and compare rhodanese activity in different tissues of adult male and female goats (Capra hircus). The results showed that the specific activity of rhodanese in different tissues was significantly different (P<0.05). The highest activity of rhodanese was in epithelium of rumen, followed by epithelia of reticulum and omasum and liver. No significant difference was observed when tissues of male and female goats were compared. The lowest specific activity of rhodanese was observed in spleen, urinary bladder, lymph node, ovary, skeletal muscle and pyloric muscle of abomasum. The results of this study may indicate the involvement of rhodanese in cyanide detoxification in goat tissues that have greater potential to be exposed to higher levels of cyanide. PMID- 12628383 TI - Molecular cloning and characterization of a cDNA encoding a novel cuticle protein in the silkworm, Bombyx mori. AB - We have cloned the full length of a novel cDNA named Bombyx mori cuticle protein that contains an AlaAlaProAla/Val-repeat (BMCPA) from a cDNA library of integument in the larval silkworm. Both a typical tandem repeat (A-A-P-A/V) for cuticle protein and a unique tandem repeat with Ser, Ala, Gly, Pro, Val, Tyr and Thr were observed in the predicted amino acid sequence of the cDNA encoding BMCPA. Approximately 80% of the amino acids in BMCPA were composed of Ser, Ala, Gly, Pro, Val and Tyr. Northern-hybridization analysis indicated that BMCPA mRNA is expressed only in the larval epidermis and that the expression pattern of the BMCPA gene in the developmental stage was observed mainly at the larval stage. We propose BMCPA may be a novel component of cuticle, and may play an important role in the integument of the larval silkworm. PMID- 12628384 TI - Amylolytic activity in fish intestinal mucosa: temperature effects. AB - The activity, temperature characteristics and energy of activation of amylolytic enzymes in the intestinal mucosa were studied in six species of fish living in a boreal zone [burbot (Lota lota L.), northern pike (Exos lucius L.), perch (Perca fluviatilis L.), bream (Abramis brama L.), roach (Rutilis rutilis L.), and carp (Cyprinus carpio L.)] and in three species from tropical and subtropical areas [pilchard (Sardina pilchardus W.), jack mackerel (Trachurus trecae C.) and round sardinella (Sardinella aurita V.)]. The amylolytic activity correlated with the feeding habits: it was essentially lower in predators. The enzyme activity at low temperature, relative to the maximal activity, was correlated with the natural environmental temperature where the species lived. At low temperature the relative activity was higher in boreal fish than in tropical and subtropical fish. We found a breakpoint in the Arrhenius plots in all fish species, except for jack mackerel. The energy of activation in predators decreased below the breakpoint in the low-temperature region. The energy of activation in benthophages of the Aral-Ponto-Caspian area was lower at higher temperatures above the breakpoint. A reduction in activation energy in the range of physiological temperatures might indicate adaptation to the environmental temperature. PMID- 12628385 TI - Cloning of Cyc (Bmal1) homolog in Bombyx mori: structural analysis and tissue specific distributions. AB - Cycle (Bmal1) is one of the circadian clock genes and the key regulator of the circadian system in many organisms, encoding a bHLH-PAS transcription factor. In the present study, we cloned cycle homolog (BmCyc) in Bombyx mori. We performed polymerase chain reaction with degenerated primers deduced from the conserved amino acid sequences of mammalian BMAL1 and Drosophila CYCLE. Then the partial clone obtained was used as a probe for screening a cDNA library constructed from pupal brains of B. mori. BmCyc is 5703 nucleotides long and encodes 700 amino acid residues. The BmCYC has bHLH, PAS A and PAS B domains, and the sequence identities for these domains are 85, 60 and 50%, to Drosophila CYCLE (dCYC) and 69, 58 and 50%, to human BMAL1 (hBMAL1), respectively. The deduced amino acid sequence of BmCYC is 37% identical to that of dCYC and 28% to hBMAL1. Northern blot analysis demonstrates that BmCyc gene was expressed in all the tissues tested, which were head, fat body, silk glands, and midgut. Also no significant day, night time-specific difference in expression of BmCyc gene in the head was detected. PMID- 12628386 TI - Investigation of the free radical scavenging activity of Ginkgo biloba L. leaves. AB - The free radical scavenging activity of methanolic, ethanolic and aqueous extracts from Ginkgo biloba leaves, has been determined by EPR (electron paramagnetic resonance) using the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging method. The investigation has also included selected constituents of G. biloba leaves, protocatechuic and p-coumaric acids, quercetin, rutin, isoginkgetin and (+)-catechin. PMID- 12628388 TI - Steroidal compounds from in vitro regenerated shoots of Datura metel. AB - Regeneration of adventitious shoots from young leaves of Datura metel is described. Shoot buds developed on MS medium with 2 mg/l and elongated on hormone free solid basal medium. The microshoots failed to produce alkaloids, but a number of steroidal compounds were detected. The C(28) sterol 3beta,24xi dihydroxy-ergosta-5,25-dienolide, regarded as the precursor of withanolides and related steroidal lactones, was identified. The withanolide 12 deoxywithastramonolide was also detected. PMID- 12628387 TI - Evaluation of antidiabetic effect of Momordica cymbalaria fruit in alloxan diabetic rats. AB - The oral treatment with the aqueous extract of Momordica cymbalaria fruit (MC) (0.5 g/kg) for 6 weeks showed a significant antihyperglycemic as well as antihyperlipidemic effects in the alloxan-induced diabetic rats. PMID- 12628389 TI - A new dilignan from Cynara cardunculus. AB - A new dilignan, 7,8-trans-7',8'-trans-7",8"-cis-7"',8"'-cis-8,8'-trans 4",4"',7',9,9",9"'-hexahydroxy-3,3',3",3"'-tetramethoxy-4,8":4',8"':5,7":5',7"' tetraoxy-7,9'-epoxy-8,8'-dilignan-7'-ol, named cynarinine (2), was isolated together with the known tracheloside (1) from the seeds of Cynara cardunculus. PMID- 12628390 TI - Central inhibitory activity of the aqueous extract of Crinum giganteum. AB - The aqueous extract of Crinum giganteum given intraperitoneally at doses of 6.25, 12.5 and 25 mg/kg in rats prolonged the duration of pentobarbital sleeping time and in mice reduced spontaneous motor activity, decreased the exploratory activity and attenuated amphetamine-induced stereotype behavior. The intraperitoneal and oral preliminary LD(50) in mice were found to be 627 and 1468 mg/kg, respectively. The data obtained suggest that the aqueous extract of C. giganteum contains some biologically active principles with sedative activity. PMID- 12628391 TI - Two new alkaloids from Hornschuchia obliqua. AB - The stem and roots of Hornschuchia obliqua provided two new alkaloids, 4,5 didehydroguadiscine and demethoxyguadiscine. Four known alkaloids, roemerine, guadiscine, liriodenine and cleistopholine were also isolated. The structures of the novel compounds were elucidated by spectroscopic analysis, including 2D NMR and MS. PMID- 12628393 TI - A non-ionic water-soluble seed gum from Ipomoea campanulata. AB - A non-ionic water-soluble galactomannan, having galactose and mannose in 2:3 molar ratio was isolated from endosperm of the seeds of Ipomoea campanulata. The seed gum was found to have linear chain of beta (1-->4) linked mannopyranosyl units with D-galactose side chains attached through alpha (1-->6) linkage to the main chain. This structure is similar to many commercial gums like Guar, Carob and Locust bean gum. Various physical properties of the gum were studied in order to explore the possibility of commercial exploitation of the seed gum. PMID- 12628392 TI - Analysis of pharmaceutically relevant taxoids in wild yew trees from Sardinia. AB - Needles from a series of wild yews (Taxus baccata L.) from Sardinia were investigated for their contents of 10-deacetyl baccatin III (DAB-III, 1), paclitaxel (Taxol) (2) and taxine (3). Despite a common geographical origin, ample variation of the taxoid profile was discovered, and several samples were surprisingly devoid of all terpenoid markers above. This finding is unprecedented within the European yew, while the general lack of taxine might rationalize the observation that most plants investigated are actively and impunently browsed by goats. PMID- 12628395 TI - Mitogenic and comitogenic activities of polysaccharides from some European herbaceous plants. AB - From the leaves of Plantago lanceolata, the green parts of Rudbeckia fulgida, the aerial parts of Salvia officinalis and the roots of Valeriana officinalis, crude polysaccharides have been isolated by extraction with water and further purified and fractionated by various techniques. The water-soluble polysaccharides obtained were examined for their immunomodulatory activities using the in vitro mitogenic and comitogenic rat thymocyte tests. The results indicate that in spite of the considerable differences in chemical composition and structural properties, the tested polysaccharides exhibited similar significant immunomodulatory properties with a particularly high adjuvans activity in the case of the Rudbeckia and Salvia polysaccharides. The pectic polysaccharide-rich complex from Valeriana was shown to also stimulate the immune function of bone marrow cells. PMID- 12628394 TI - Anti-inflammatory and antinociceptive activities of Loasa speciosa in rats and mice. AB - In the range of doses of 250-500 mg/kg (given i.p.) the aqueous extract of Loasa speciosa leaves showed an inhibitory effect on leukocyte migration, and a reduction on the pleural exudate, as well as dose-dependent anti-inflammatory and peripheral antinociceptive activities. PMID- 12628396 TI - Production of anthocyanins by Catharanthus roseus. AB - A stable cell suspension line of Catharanthus roseus producing anthocyanin was obtained. In this strain it was found that approximately 30% of cells regularly accumulated these metabolites and that anthocyanin accumulation occurred between the second half of log phase and the stationary phase of the culture growth cycle. The anthocyanins in the suspension cultures were compared with those biosynthetized in the flowers both of regenerated by somatic embryogenesis and field-grown plants. Six anthocyanins were identified in all the examined samples, three 3-O-glucosides and three 3-O-(6-O-p-coumaroyl) glucosides of petunidin, malvidin and hirsutidin. The hirsutidin coumaroyl glucoside has not been reported previously, and was predominat in all samples. The anthocyanin relative content was similar for cell suspensions and flowers from regenerated plants but different from field-grown plant flowers; instead, the total content was almost the same for the two flower types and higher compared to suspension culture content. PMID- 12628397 TI - Quantitative HPLC analysis of withanolides in Withania somnifera. AB - One of the most widely used herbs in Ayurvedic medicine is Ashwaghanda, Withania somnifera, a shrub commonly found on the Indian subcontinent. As this plant is increasingly becoming a popular adaptogenic in the western world, analytical methods for its identification and quality control are in demand. Thus, a HPLC method for the determination of withaferin A and withanolide D was developed. The system was successfully used to investigate the presence of the markers in different W. somnifera plant parts as well as to analyze their content in market products. PMID- 12628398 TI - Effect of meliacine, a plant derived antiviral, on tumor necrosis factor alpha. AB - The effect of meliacine (MAS) and two fractions MAB 1 and MAB 2 obtained from it on the in vitro production of TNF-alpha of murine macrophages induced by bacterial lipopolysaccharide (LPS) (from Escherichia coli) was tested. Simultaneous administration of the above fractions (ranging from 14 to 56 microg/ml) and LPS (10 microg/ml) to a macrophage culture significantly increased the amount of TNF-alpha released at 24 h of induction in a dose-dependent manner. Meliacine alone, at a concentration of 56 microg/ml, is a weak inducer of TNF alpha production. PMID- 12628399 TI - Effects of Bauhinia megalandra aqueous leaf extract on intestinal glucose absorption and uptake by enterocyte brush border membrane vesicles. AB - Aqueous extract of Bauhinia megalandra leaves was able to inhibit the intestinal glucose absorption in a concentration-dependent way and additive to phlorizine. Moreover, B. megalandra leaf extract drastically reduced the 14C-glucose uptake by enterocyte brush border membrane vesicles. The B. megalandra leaf extract administrated orally, simultaneously with glucose improved the glucose tolerance with a significant reduction of the 30-min peak. The extract did not have an effect on the glucose tolerance when glucose was administrated subcutaneously. PMID- 12628401 TI - Ethnobotanical knowledge of local Arab practitioners in the Middle Eastern region. AB - An ethnobotanical survey was conducted among 31 local Arab practitioners living in Galilee, Negev, Golan Heights and West Bank. Seeing that the Arabic traditional medicine is in danger of disappearing, a multilevel program is required, involving the training of local practitioners, an establishment of a regional medicinal plant botanical garden and a field gene bank for plant preservation. PMID- 12628400 TI - Antioxidant activity of Paraguayan plant extracts. AB - The antioxidant properties of six medical herbs used in the traditional Paraguayan medicine were studied using free radical-generating systems. The methanol extracts from Aristolochia giberti, Cecropia pachystachya, Eugenia uniflora, Piper fulvescens, Schinus weinmannifolia and Schinus terebinthifolia protected against enzymatic and non-enzymatic lipid peroxidation in microsomal membranes of rat. C. pachystachya, E. uniflora, S. weinmannifolia and S. terebinthifolia showed the highest scavenging activity on the superoxide and DPPH radicals. PMID- 12628402 TI - Cytoprotective effect of piperine against benzo[a]pyrene induced lung cancer with reference to lipid peroxidation and antioxidant system in Swiss albino mice. AB - The cytoprotective effect of piperine on benzo[a]pyrene (B[a]P) induced experimental lung cancer was investigated in male Swiss albino mice. Oral administration of piperine (100 mg/kg body wt.) effectively suppressed lung cancer initiated with B[a]P as revealed by the decrease in the extent of lipid peroxidation with concomitant increase in the activities of enzymatic antioxidants (superoxide dismutase, catalase and glutathione peroxidase) and non enzymatic antioxidant (reduced glutathione, vitamin E and vitamin C) levels when compared to lung cancer bearing animals. Our data suggest that piperine may extend its chemopreventive effect by modulating lipid peroxidation and augmenting antioxidant defense system. PMID- 12628403 TI - Antidiarrhoeal activity of the bark extract of Careya arborea Roxb. AB - The methanol extract of the Careya arborea Roxb. bark significantly reduced castor oil-induced diarrhoea in mice. This effect supports the local traditional use of the plant against diarrhoea. PMID- 12628404 TI - Antinociceptive activity of the aerial parts of Solanum xanthocarpum. AB - The methanolic extract of Solanum xanthocarpum aerial parts, given orally at 125, 250 and 500 mg/kg, showed significant antinociceptive activity in mice. PMID- 12628406 TI - Cytotoxic activity and quality control determinations on Chelidonium majus. AB - The quality control determinations on herba and radix Chelidonii prepared from Chelidonium majus L., growing wildly in Bursa, Uludag surroundings, have been conducted according to DAB 9. Ash, humidity and total alkaloidal content have been carried out. LC(50) value of chelidonine and protopine, the most important alkaloids, and the alcoholic and water extracts of herba Chelidonii were determined using brine shrimp (Artemia salina) lethality bioassay. PMID- 12628405 TI - Antimicrobial activity of Ximenia americana. AB - Extracts of the bark, leaves, root and stem of Ximenia americana were tested for their antimicrobial and antifungal activity. PMID- 12628407 TI - Antimicrobial activity of the leaves and flowering tops of Acanthospermum hispidum. AB - The ethanolic extracts of the leaves and flowering tops of Acanthospermum hispidum showed varying degrees of activity against a wide range of pathogenic bacteria. The activity resided mostly in the polar fractions of the alcoholic extract, being only slight in the non-polar fractions. No activity was observed for the aqueous extract of the fresh plant material. PMID- 12628408 TI - Antibacterial and free radical scavenging activity of the seeds of Agrimonia eupatoria. AB - n-Hexane, dichloromethane and methanol extracts of the seeds of Agrimonia eupatoria have been assessed for antibacterial and free radical scavenging activity. PMID- 12628409 TI - Antimicrobial activity of the leaves and seeds of Bixa orellana. AB - The ethanolic extracts of the leaves and seeds of Bixa orellana showed a broad spectrum of antimicrobial activity. The activity of the leaf extract was more pronounced. PMID- 12628410 TI - Evaluation of the anti-microbial and anti-inflammatory activities of the medicinal plants Dodonaea viscosa, Rumex nervosus and Rumex abyssinicus. AB - The crude extracts of the leaves of Dodonaea viscosa and Rumex nervosus as well as of the root of Rumex abyssinicus were tested for anti-microbial and anti inflammatory activities. It was observed that the three plants possess antibacterial activity against Streptococcus pyogenes and Staphylococcus aureus and strong activity against Coxsackie virus B3 and influenza A virus. In contrast, none of them exhibited anti-fungal activity. The anti-inflammatory activity test results verified that only R. abyssinicus inhibited the synthesis of prostaglandin (PG) E(2). PMID- 12628411 TI - Preliminary assay on the antioxidative activity of Maydis stigma extracts. AB - Antioxidative effects of Maydis stigma methanolic extracts were studied on the level of lipid peroxidation in liposomes, induced by Fe(2+)/ascorbate system and measured spectrophotometrically by the TBA-test. The most significant inhibition of lipid peroxidation was observed when methanolic extracts of fully developed, mature corn silk were applied. The same test, performed after fractionation of the most active extract, showed that most of the activity was concentrated in fractions with moderate lipophilicity, containing phenolic acids, flavonoid aglyca and resembling monosides. PMID- 12628412 TI - Antibacterial activity of Vernonia cinerea. AB - The benzene extract of Vernonia cinerea showed a broad spectrum of antibacterial activity. PMID- 12628413 TI - Bioactivity of gentiopicroside from the aerial parts of Centaurium erythraea. AB - Gentiopicroside (1), a secoiridoid glycoside isolated from the methanol extract of the aerial parts of Centaurium erythraea, has been assessed for antibacterial and free radical scavenging activities. General toxicity of 1 has also been determined by brine shrimp lethality bioassay. PMID- 12628414 TI - Antimicrobial activity of Toona ciliata and Amoora rohituka. AB - The extracts of two Bangladeshi medicinal plants, Toona ciliata (stem bark) and Amoora rohituka (stem bark), along with siderin, a major coumarin from T. ciliata, exhibited significant in vitro antibacterial activity. The extracts also demonstrated mild antifungal effect. PMID- 12628415 TI - Antibacterial activity and cytotoxicity of extractives from Uvaria hamiltonii stem bark. AB - The extracts as well as piperolactum C, goniopedaline, 6beta-hydroxystigmasta 4,22-dien-3-one and a mixture of cis- and trans-4-hydroxymelleins obtained from Uvaria hamiltonii stem bark exhibited mild to moderate in vitro antibacterial activity. The results obtained were compared with a standard antibiotic, kanamycin. In a brine shrimp lethality bioassay the pet-ether, dichloromethane, methanol extracts and a major alkaloid, piperolactum C were found to exhibit mild to moderate cytotoxicity. PMID- 12628416 TI - Antifungal activities of the leaves of three Pistacia species grown in Turkey. AB - The crude extracts obtained from the leaves of Pistacia vera, Pistacia terebinthus and Pistacia lentiscus were tested for antifungal activities against three pathogenic agricultural fungi, Phythium ultimum, Rhizoctania solani and Fusarium sambucinum. The extracts significantly inhibited the growth of P. ultimum and R. solani. However, the antifungal activity was not observed against F. sambucinum. PMID- 12628417 TI - Content of metals in the seeds of Xanthium spinosum and Xanthium italicum. AB - The quantitative analysis of metallic cations in the seeds of Xanthium spinosum L. and Xanthium italicum Moretti was performed. The differences for the two species reflect the different presence of some organic compounds having a ligand character. PMID- 12628418 TI - Chemical composition and antifungal properties of essential oils of three Pistacia species. AB - The chemical composition of essential oils obtained from the leaves of Pistacia vera, Pistacia terebinthus, Pistacia lentiscus and the resin of Pistacia lentiscus were analyzed by GC and GC-MS. alpha-Pinene, beta-pinene, limonene, terpinen-4-ol and alpha-terpineol were found to be the major components. The antifungal activities of the above oils and P. lentiscus resin (total, acidic and neutral fractions) against the growth of three agricultural pathogens, Pythium ultimum, Rhizoctonia solani and Fusarium sambucinum were evaluated. Some doses of P. terebinthus, P. vera and P. lentiscus leaf oils and total and neutral fraction of P. lentiscus resin significantly inhibited the growth of R. solani. However, all samples did not show antifungal activity against P. ultimum and F. sambucinum, but increased the growth of F. sambucinum. PMID- 12628419 TI - Antiviral lanostanoid triterpenes from the fungus Ganoderma pfeifferi. AB - Ganodermadiol, lucidadiol and applanoxidic acid G were isolated as first triterpenes from the European Basidiomycete Ganoderma pfeifferi. The compounds show antiviral activity against influenza virus type A and HSV type 1. PMID- 12628420 TI - Amides from Piper tuberculatum fruits. AB - The fruits of Piper tuberculatum Jacq yielded piperine S (1) and N-(12,13,14 trimethoxydihydrocinnamoyl)-delta(3)-piperidin-2-one (2). Their complete NMR analysis based on one- and two-dimensional experiments is reported. PMID- 12628421 TI - Bioactive constituents of Leptadenia arborea. AB - The aerial part of Leptadenia arborea has been shown to contain pinoresinol (1), syringaresinol (2), leucanthemitol (3) and E-ferulaldehyde (4). These known compounds are being reported for the first time from this plant. Among them, syringaresinol has shown an inhibitory effect against acetylcholinesterase. The IC(50) (the concentration of 50% enzyme inhibition) value of this compound was 200 microg/ml. PMID- 12628422 TI - A new flavonoid from Limnophila rugosa. AB - A minor flavonoid constituent of Limnophila rugosa has been isolated from its petrol extract and identified as 5,7-dihydroxy-8,3',5'-trimethoxyflavone (1). PMID- 12628423 TI - Alkaloids of Zanthoxylum budrunga. AB - Pseudophrynamine, lunacridine and the new compound 2-(2',4',6'-trimethyl heptenyl)-4-quinozolone were isolated from the leaves of Zanthoxylum budrunga. PMID- 12628424 TI - A new dicoumarinyl ether and two rare furocoumarins from Ruta montana. AB - A new dicoumarinyl ether, rutamontine and two rare furocoumarins, heraclenol and isopimpinellin, were isolated from Ruta montana. The structure of rutamontine as 6-hydroxy-7-methoxy-3,7'-dicoumarinylether was determined by spectroscopic methods. PMID- 12628427 TI - Micromechanics/structure relationships in the human mandible. AB - OBJECTIVES: A clear understanding of the relationship between the micromechanical properties and orientation of the osteons within the mandible is important to understand mandibular function, fracture repair, treatment of temporo-mandibular joint disorders, the materials and organization of dental implants. The objective of this research was to obtain the micromechanical properties of human mandibular cortical bone as a function of orientation from TMJ to TMJ. METHODS: A mandible obtained from a deceased 66 year-old female free of bone disease was used. The mandible was embedded in polymethylmethacrylate. The micromechanical properties analysis was obtained using the UH3 scanning acoustic microscope (SAM; Olympus Co., Tokyo, Japan). The coordinates system is defined such that the inferior border of mandibular is positioned on the x-y plane. x is along the anterior posterior direction, y is in the horizontal direction and z is in superior inferior direction. RESULTS: The osteonal orientations were almost parallel to the x axis and eventually branched into two directions towards the coronoid process and condylar head. The SAM revealed that almost the whole area of the mandible body was found to be transversely isotropic in the plane perpendicular to the x axis. In the parallel and oblique directions, all data were transversely isotropic with respect to the x axis. Data of the perpendicular osteons were transversely isotropic with respect to the z axis. SIGNIFICANCE: Having actual micromechanical properties as a function of orientation in the mandible could provide base line data for: fracture repair; choice of bone replacement materials. PMID- 12628426 TI - Mapping of tooth deformation caused by moisture change using moire interferometry. AB - Bonding of composite restorative materials to teeth usually involves surface etching and drying of both dentin and enamel. These two hard tissues have very different water contents. OBJECTIVES: The purpose of this study was to evaluate the effects of changes in humidity on the dimensional changes in dentin disks constrained by enamel and in unconstrained dentin. METHODS: Changes in humidity were used to induce changes in the moisture level of the hard tissues, which created a mechanical load across the dentin-enamel junction (DEJ). High sensitivity moire interferometry was used to measure the magnitudes of the strains. Diffraction gratings (with a sensitivity of 2400 lines/mm) were transferred to moist polished sections of human teeth, which were subsequently dehydrated and then hydrated again. The fringe patterns from moire interferometry recorded the change in deformation between the moisture states. RESULTS: The results indicated that there were wide variations in strain between the two specimen geometries. The strains were fairly constant through the bulk of the dentin in both specimen types. However, the strain gradients were very high across the dentin-enamel interface into the dentin in the unconstrained geometry, and nearly zero in the constrained geometry. SIGNIFICANCE: Moire interferometry is a powerful tool to study the deformation of materials that are not isotropic and are not linearly elastic. It provides full-field, high-resolution deformation fields in tooth specimens. Deformations observed in this study indicate that the DEJ zone is a unique material interface that needs to be better understood in terms of normal tooth function, and that its material properties be taken into account when natural hard tissues are restored. PMID- 12628428 TI - Mechanical properties and microstructures of cast Ti-Cu alloys. AB - OBJECTIVES: This study evaluated the mechanical properties of cast Ti-Cu alloys with the hope of developing an alloy for dental casting with better mechanical properties than unalloyed titanium. METHODS: Ti-Cu alloys with five concentrations of copper (0.5, 1.0, 2.0, 5.0 and 10.0 mass%) were made in an argon-arc melting furnace. The alloys were cast into magnesia-based molds using a centrifugal casting machine. The microstructure, microhardness profile of the specimen cross section, tensile strength, yield strength, and elongation were determined for the castings. Scanning electron microscope fractography was undertaken for the fractured surfaces after tensile testing. XRD was performed on the polished specimens. Results were analyzed using one-way ANOVA and the Student Newman-Keuls tests. RESULTS: The mean tensile strengths of all the cast Ti-Cu alloys were significantly (p<0.05) higher than for cast commercially pure titanium (CP Ti). Of the Ti-Cu alloys tested, the 5 and 10% Cu alloys had significantly higher strength than the rest. The 10% Cu alloy exhibited the lowest mean elongation. CP Ti and the 0.5 and 1% Cu alloys showed higher ductility. The bulk hardness of all the cast Ti-Cu alloys, except for the 10% Cu alloy, and CP Ti was approximately the same. SIGNIFICANCE: By alloying with copper, the cast titanium became stronger. Increases in the tensile strength (30%) and yield strength (40%) over CP Ti were obtained for the 5% Cu alloy. Elongation was approximately 3%, which was similar to cast Ti-6Al-4V. Ti-Cu alloys, such as the 5% Cu alloy, could be used for prosthetic dental applications if other properties necessary for dental castings are obtained. PMID- 12628429 TI - Stickiness prior to setting of some light cured resin-composites. AB - OBJECTIVES: To describe and evaluate a method for deriving a laboratory measure of stickiness, and to compare a set of dental resin-composites by this procedure. MATERIALS AND METHODS: Twelve commercial resin composites were selected. Each material was placed in a cylindrical mold (phi=6.1 x 2.2 mm(2) depth) held either at 23 or at 37 degrees C. A flat-ended stainless-steel instrument was placed onto the surface of the unset sample with a force 350 g. After 2s it was moved vertically at 2 cm/s. This caused a sticky composite to be elevated to a maximum height between 0.2 and 1.8 mm until detachment from the instrument occurred. The elevated material was immediately solidified by light curing at 600 mW/cm(2) for 40s. These elevated profiles were mapped for both height and projected area of elevation. An instrument was constructed for this purpose in our laboratories. Both the projected heights and areas were evaluated as potential measures of stickiness. One-way ANOVA and Duncan's multiple range test were used for statistical analysis. RESULTS: Projected heights ranged from 0.2 to 1.8 mm and projected areas of elevation ranged from 4.0 to 23.9 mm(2). The greatest values were found for microfine anterior composites (Silux plus, Filtek A110). There was a significant difference (p<0.05) between the material (SX) with the highest stickiness and all of the others. The set of profile height and area values were found to be strongly correlated (r(2)=0.9600). SIGNIFICANCE: Once inserted and the load removed, the ideal material stays in place. This is generally assisted by an increase in viscosity and a decrease in stickiness. The stickiness test described is original and may prove to be useful as a standard test method for characterizing the handling stickiness of resin composite materials. PMID- 12628430 TI - In vitro corrosion behavior of bioceramic, metallic, and bioceramic-metallic coated stainless steel dental implants. AB - OBJECTIVES: The most common metals and alloys used in dentistry may be exposed to a process of corrosion in vivo that make them cytotoxic. The biocompatibility of dental alloys is primarily related to their corrosion behavior. The aim of this work was to evaluate the corrosion behavior and thus the biocompatibility of the uncoated and coated stainless steels and compare the effect of type of coatings on corrosion behavior. METHODS: Three types of coatings, hydroxyapatite (HA), titanium (Ti), and a double-layer HA/Ti on AISI 316L stainless steel were made. HA coating was produced using plasma-spraying technique and Ti coating was made using physical vapor deposition process. In order to perform a novel double-layer composite coating, a top layer of HA was plasma-sprayed over a physical vapor deposited Ti layer on AISI 316L stainless steel. Structural characterization techniques including XRD, SEM and EDX were used to investigate the microstructure, morphology and crystallinity of the coatings. Electrochemical potentiodynamic tests were performed in physiological solutions in order to determine and compare the corrosion behavior of the coated and uncoated specimens as an indication of biocompatibility. RESULTS: Double-layer HA/Ti coating on AISI 316L SS had a positive effect on improvement of corrosion behavior. The decrease in corrosion current densities was significant for these coated specimens and was much lower than the values obtained for uncoated and single HA coated specimens. Ti coating on AISI 316L SS also has a beneficial effect on corrosion behavior. The results were compared with the results of corrosion behavior of HA coated commercially pure titanium (cpTi) and uncoated cpTi. SIGNIFICANCE: These results demonstrated that the double-layer HA/Ti coated 316L SS can be used as an endodontic implant and two goals including improvement of corrosion resistance and bone osteointegration can be obtained simultaneously. PMID- 12628431 TI - Microtensile bond strength between adhesive cements and root canal dentin. AB - OBJECTIVES: The hypotheses tested were that the bond strength of adhesive cements to root canal dentin (1) would be reduced as a function of configuration factor, polymerization process and type of luting material and (2) would be lowered near the apex of the tooth. METHODS: Human canines and premolars were prepared for post cementation using Single Bond/Rely X ARC, ED Primer/Panavia F, C and B Metabond, and Fuji Plus. The specimens were divided into two groups. For intact roots, the posts were luted using standard clinical procedures. For flat roots, the posts were applied directly into flat ground canals. All roots were sectioned into 0.6 mm thick slices, trimmed mesio-distally and stressed to failure at 1 mm/min. The muTBS of each slab was calculated as the force at failure divided by the bonded cross-sectional surface area. The results were compared using a one way ANOVA and Tukey multiple comparison intervals (alpha=0.05). Least squares linear regression analysis was used to assess the effect of dentin location on bond strength. RESULTS: All cements showed significantly (p0.05). For the SG3 material no significant differences were seen between the as-cast glass and cerammed discs (p>0.05), with the WMC investment producing the smoothest discs (p<0.05), and the TG investment the roughest (p<0.05). The LG112 material showed a significant difference between glass and cerammed discs for the gypsum-bonded investments (p<0.05) but not when using phosphate-bonded investments (p>0.05). With LG112 the WMC investment produced smoother discs than the DC and TG investments (p<0.05). The DC investment produced rougher discs than the two phosphate-bonded investments (p<0.05). SIGNIFICANCE: The selection of investment material can have a significant effect on the as-cast surface finish of castable glass-ceramic materials. A gypsum-bonded investment material gave smoother surface finishes compared to phosphate-bonded investments. The ceramming process significantly increases surface roughness. PMID- 12628435 TI - Static and cyclic loading of fiber-reinforced dental resin. AB - OBJECTIVE: The aim of this study was to evaluate the flexure strength of unidirectional fiber-reinforced resins under static and cyclic loading with and without thermal cycling. METHODS: The fiber-reinforced resin materials chosen for this project were commercially available endodontic posts and commercially procured bar samples. For all materials, controls for flexure strength were tested in air and in water using three-point loading. Specimens were thermal cycled between 7 and 63 degrees C for 6000 cycles. A staircase approach was used to determine the flexure fatigue limit and scanning microscopy was used to examine the microstructure. RESULTS: The carbon/graphite fiber-reinforced resin posts and the glass FiberKor posts were significantly stronger than the ceramic (zirconia) and the other glass-reinforced resin materials. Thermal cycling caused a significant lowering (11-24%) of the flexure strength for each resin based post system. The ceramic post system decreased only by 2%. Further, for standard size glass fiber-reinforced resin bars, no significant differences between testing in air and water was observed, but a significant difference between static and cyclic loading was noted. SIGNIFICANCE: The decreases in the strength property due to thermal cycling and the cyclic loading of these materials indicates that their utilization in the oral environment enhances their degradation, and potentially shortens their clinical life. PMID- 12628436 TI - Evaluation of the long-term corrosion behavior of dental amalgams: influence of palladium addition and particle morphology. AB - OBJECTIVES: The purpose of this investigation was to evaluate the long-term corrosion behavior of experimental amalgams as a function of particle morphology and palladium content. METHODS: Samples of four experimental high copper amalgams were prepared according to ADA specifications. Two of them had the same chemical composition but one had lathe cut particles (LCP) and the other had spherical particles (SP). The two others had spherical powders with an addition of 0.5 wt% of palladium (SP 0.5) and 1 wt% of palladium(SP 1) for the other. Corrosion resistance was evaluated by electrochemical techniques in Ringer's solution in a thermostated cell at 37 degrees C for samples aged 5, 8, 12, 16 months and 10 years. Potentiokinetic curves were drawn and the potential and the current density corresponding to the first anodic peak were registered. RESULTS: For all the amalgam samples the corrosion behavior improves over the 10-year period. SP samples exhibit a better behavior than LCP. Palladium addition improves corrosion behavior as compared to samples without palladium. No real difference is found regarding the amount of palladium between 0.5 and 1%. The potentials progress from a range between 0 and 20 mV/SCE to a range of 60-80 after 10 years. The stabilization of the potential begins after only 16 months. Except for the LCP, all the values converge to the same level of 80 mV/SCE. SIGNIFICANCE: The addition of no more than 0.5 wt% Palladium in a high copper amalgam powder improves the corrosion behavior of the amalgam up to a period of 10 years. The potential of the first anodic peak increases for each amalgam, probably in relation to the evolution of the structure of the material. Clinically, it is of interest to consider the good electrochemical behavior of older restorations when contemplating the repair or replacement of such fillings. At the same time, galvanic current can occur when a new amalgam restoration is placed in contact with an old one even if the same amalgam is used. In this situation, the new filling will be anodic and its degradation will be accelerated. The evaluation of the corrosion behavior of dental amalgams has to take into account the age of the samples. PMID- 12628437 TI - Erosion of water-based cements evaluated by volumetric and gravimetric methods. AB - OBJECTIVES: To compare the erosion of glass ionomer, zinc phosphate and polycarboxylate cements using volumetric and gravimetric methods. METHODS: For the volumetric method, the eroded depth of cement placed in a cylindrical cavity in PMMA was measured using a dial gauge after immersion in an eroding solution. For the gravimetric method, the weight of the residue of a solution in which a cylindrical specimen had been immersed was measured. 0.02 M lactic acid solution (0.02 M acid) and 0.1 M lactic acid/sodium lactate buffer solution (0.1 M buffer) were used as eroding solutions. The pH of both solutions was 2.74 and the test period was 24 h. RESULTS: Ranking of eroded depth and weight of residue was polycarboxylate>zinc phosphate>glass ionomers. Differences in erosion were more clearly defined by differences in eroded depth than differences in weight of residue. In 0.02 M acid, the erosion of glass ionomer using the volumetric method was effected by the hygroscopic expansion. In 0.1 M buffer, the erosion for polycarboxylate and zinc phosphate using the volumetric method was much greater than that using the gravimetric method. This is explained by cryo-SEM images which show many holes in the surface of specimens after erosion. It appears that zinc oxide is dissolved leaving a spongy matrix which easily collapses under the force applied to the dial gauge during measurement. SIGNIFICANCE: The volumetric method that employs eroded depth of cement using a 0.1 M buffer solution is able to quantify erosion and to make material comparisons. PMID- 12628438 TI - Determination of residual double bonds in resin-dentin interface by Raman spectroscopy. AB - OBJECTIVES: The quality of the hybrid layer is believed to be more important than the thickness of this layer. The purpose of this study was to investigate a method to analyze the percentage of adhesive resin residual double bonds in the dentin-resin interface using laser Raman spectroscopy. METHODS: Bovine dentin was treated with dentin adhesives and resin composite was bonded according to the manufacturers' instructions. The specimens were sectioned parallel to dentinal tubules and the surfaces were then polished to 1 microm diamond pastes. Raman spectra were recorded along a line perpendicular to the dentin-resin interface in steps of 0.2 microm. The measurement of residual C=C bond was made on a relative basis by comparing the C=C unpolymerized methacrylate stretching vibration (1638 cm(-1)) against the C=O stretching mode of the ester group (1719 cm(-1)). The percentage of residual double bonds including pendant and monomeric double bonds was calculated by comparing the obtained ratio with that of uncured adhesive resin. RESULTS: The amount of residual double bonds in the hybrid layer varied from 10 to 25% compared to the uncured adhesives, a relatively higher percentage was detected for Fluoro Bond (12.3-23.6%) and Single Bond (9.5-21.8%), and lower for Mac Bond II (10.6-18.0%) and Mega Bond (10.7-16.3%). No relationship was seen between the percentage of remaining double bonds and the location within the resin-dentin interface. SIGNIFICANCE: Laser Raman microscopy used was a useful tool for measuring the residual double bonds in the dentin-resin interface. PMID- 12628439 TI - Melatonin receptors and their regulation: biochemical and structural mechanisms. AB - There is growing evidence demonstrating the complexity of melatonin's role in modulating a diverse number of physiological processes. This complexity could be attributed to the fact that melatonin receptors belong to two distinct classes of proteins, that is, the G-protein coupled receptor superfamily (MT(1), MT(2)) and the quinone reductase enzyme family (MT(3)) which makes them unique at the molecular level. Also, within the G-protein coupled receptor family of proteins, the MT(1) and MT(2) receptors can couple to multiple and distinct signal transduction cascades whose activation can lead to unique cellular responses. Also, throughout the 24-hour cycle, the receptors' sensitivity to specific cues fluctuates and this sensitivity can be modulated in a homologous fashion, that is, by melatonin itself, and in a heterologous manner, that is, by other cues including the photoperiod or estrogen. This sensitivity of response may reflect changes in melatonin receptor density that also occurs throughout the 24-hour light/dark cycle but out of phase with circulating melatonin levels. The mechanisms that underlie the changes in melatonin receptor density and function are still not well-understood, but data is beginning to show that transcriptional events and G-protein uncoupling may be involved. Even though this area of research is still in its infancy, great strides are being made everyday in elucidating the mechanisms that underlie melatonin receptor function and regulation. The focus of this review is to highlight some of these discoveries in an attempt to reveal the uniqueness of the melatonin receptor family while at the same time provide thought-provoking ideas to further advance this area of research. Thus, a brief overview of each of the mammalian melatonin receptor subtypes and the signal transduction cascades to which they couple will be discussed with a greater emphasis placed on the mechanisms underlying their regulation and the domains within the receptors essential for proper signaling. PMID- 12628440 TI - Increased intracellular calcium activates serum and glucocorticoid-inducible kinase 1 (SGK1) through a calmodulin-calcium calmodulin dependent kinase kinase pathway in Chinese hamster ovary cells. AB - SGK1 is one of the protein-serine/threonine kinases that is activated by insulin in a PI3K-dependent manner. Although SGK1 mediates a variety of biological activities, the mechanisms regulating its activity remain unclear. In this study, we examined the potential roles of calcium signaling in the activation of SGK1. Treatment of CHO-IR cells with a cell-permeable calcium chelator, BAPTA-AM, abolished the insulin-induced activation of SGK1. Increasing intracellular calcium concentration by treating cells with thapsigargin or ionomycin induced a 6-8 fold increase in SGK1 activation. This was not affected by a PI3K inhibitor, wortmannin, but was completely inhibited by the calmodulin inhibitors, W 7 and W 5. Co-transfection of CHO cells with FLAG-SGK1 and CaMKK revealed the direct association of CaMKK with SGK1. These results suggest a calcium-triggered signaling cascade in which an increase in intracellular calcium concentration directly stimulates SGK1 through CaMKK. PMID- 12628441 TI - ETA endothelin receptors are involved in the ouabain-induced haemodynamic effects in the periaqueductal gray area of rats. AB - The aim of the present study was to asses the effects on blood pressure and vascular resistances elicited by microinjections of ouabain (OUA) within the periaqueductal gray area (PAG). We also tested whether peripheral vascular responses caused by exogenous intra-PAG ouabain involve activation of the PAG endothelin system. In normotensive Sprague-Dawley rats the basal mean arterial blood pressure (MABP) was 114 +/- 3 mmHg. This was significantly increased by OUA (3 micro g, 122 +/- 2 mmHg, p < 0.05; and 6 micro g, 139 +/- 3 mmHg, p < 0.01) microinjected into the PAG area. Increases in MABP were associated with increases in total peripheral resistances (TPR), organ vascular resistances, and with reduced blood flow of almost all the organs tested: kidneys, skeletal muscle, skin, stomach, spleen, testes and intestine. Cardiac output did not change. Changes in the above vascular parameters induced by OUA were significantly (p < 0.01) reduced by intra-PAG microinjections of FR139317 (a selective ETA receptor antagonist, 5 nmol), SB209670 (a non-selective ETA/ETB receptor antagonist, 3 nmol), but not by BQ 788 (a selective ETB receptor antagonist, 5 nmol). In conclusion, OUA into the PAG area of normotensive rats caused significant changes in peripheral vascular parameters that are reduced by ETA receptor antagonists. These results indicate that PAG-ET-1 system via an action on ETA receptors is involved in the OUA effects. PMID- 12628442 TI - Distribution of [14C]-trans-resveratrol, a cancer chemopreventive polyphenol, in mouse tissues after oral administration. AB - Trans-resveratrol, a phenolic compound present in wine, has been reported to be a potential cancer chemopreventive agent. However, although it has numerous biological activities in vitro, there are few data about its bioavailability and tissue distribution in vivo. The objectives of this study were to investigate the absorption and tissue distribution of 14C-trans-resveratrol following oral administration to mice. Male Balb/c mice were given a single oral dose of 14C trans-resveratrol and were sacrificed at 1.5, 3 or 6 h postdose. The distribution of radioactivity in tissues was evaluated using whole-body autoradiography, quantitative organ-level determination and microautoradiography. In addition, identification of radioactive compounds in kidney and liver was done with high performance liquid chromatography. Autoradiographic survey of mice sections as well as radioactivity quantification in various organs revealed a preferential fixation of 14C-trans-resveratrol in the organs and biological liquids of absorption and elimination (stomach, liver, kidney, intestine, bile, urine). Moreover, we show that 14C-trans-resveratrol derived radioactivity is able to penetrate the tissues of liver and kidney, a finding supported by microautoradiography. The presence of intact 14C-trans-resveratrol together with glucurono- and/or sulfoconjugates in these tissues was also shown. This study demonstrates that trans-resveratrol is bioavailable following oral administration and remains mostly in intact form. The results also suggest a wide range of target organs for cancer chemoprevention by wine polyphenols in humans. PMID- 12628443 TI - Histamine-releasing reaction induced by the N-terminal domain of Vibrio vulnificus metalloprotease. AB - A zinc metalloprotease secreted by Vibrio vulnificus, an opportunistic human pathogen causing septicemia and wound infection, stimulates exocytotic histamine release from rat mast cells. This protease consists of two functional domains: the N-terminal domain that catalyzes proteolytic reaction and the C-terminal domain that promotes the association with a protein substrate or cell membrane. Like the intact protease, the N-terminal domain alone also induced histamine release from rat peritoneal mast cells in a dose- and time-dependent manner. However, the reaction induced was apparently weak and went on more slowly. The nickel-substituted protease or its N-terminal domain, each of which has the reduced proteolytic activity due to decreased affinity to a substrate, showed much less histamine-releasing activity. When injected into the rat dorsal skin, the N-terminal domain also evoked enhancement of the hypodermic vascular permeability, while the activity was comparable to that of the protease. Taken together, the protease may stimulate histamine release through the action of the catalytic center of the N-terminal domain on the target substance(s) on the mast cell membrane. The C-terminal domain may support the in vitro action of the N terminal domain by coordination of the association of the protease with the membrane, but it may not modulate the in vivo action. PMID- 12628444 TI - Differential responses from seven mammalian cell lines to the treatments of detoxifying enzyme inducers. AB - Cell-based models have been used extensively in screening novel bioactive chemical entities. In this study, seven well-established mammalian cell lines, which have different origins, were utilized to compare their responses to the treatments of three detoxifying enzyme inducers, tert-butylhydroquinone (tBHQ), beta-naphthoflavone (beta-NF), and sulforaphane (SUL), which are potential chemopreventive compounds. The enzymatic activities of glutathione s-transferase (GST), NAD(P)H:quinone oxidoreductase (QR), aldehyde reductase (AR), and glutathione reductase (GR) were measured by kinetics methods using UV-Vis spectroscopy, and analyzed statistically by Student's t-test. Among these mammalian cell lines, the mouse hepatoma Hepa1c1c7 cells were the most robust and sensitive cells, which had higher basal as well as upregulated enzymatic activities. In human cell lines, the prostate LNCaP and hepatic HepG2 cells were also very responsive to the inducers. The results suggested that different cell lines responded differently to individual detoxifying gene inducer, and the selection of appropriate cell line is important for screening potential chemopreventive agents. PMID- 12628445 TI - Heavy load exercise induced dysfunction of immunity and neuroendocrine responses in rats. AB - To determine whether immunity and neuroendocrine system is altered by different loads of exercise training in rats, eight-week-old male Sprague-Dawley rats were randomly assigned to one of the three groups: 1) cage control group (CCG); 2) moderate load training (MLT) (swimming at the intensity of 1.4 m/sec water flowing for 60 min per day); 3) heavy load training (HLT) (swimming at the intensity of 1.8 m/sec water flowing for 120 min per day). MLT and HLT rats were assigned to swim for 6 days per week for total of 6 weeks. All rats were sacrificed 36 h after their last training session. Splenocytes were pooled for assay of cell proliferation and neuropeptide contents in the hypothalamus, hypophysis and plasma were determined by radioimmunoassay while glucocorticoid specific binding in intact thymus was measured by radioligand binding assay. All rats were weighed weekly. The results showed that after 6-week training, rat splenocyte proliferation in response to Con A and LPS decreased in HLT rats compared with MLT and CCG rats. In addition, the contents of beta-endorphin, dynorphin A, arginine vasopressin and oxytocin in the hypothalamus, hypophysis and plasma were altered by HLT, as shown by increased plasma concentration of glucocorticoids and decreased glucocorticoids specific binding in intact thymus compared with MLT and CCG. Furthermore, a decreased body mass in HLT rats has been observed. The body mass of HLT rats was significantly lower than that in CCG and MLT rats at the end of the swimming training period. These data suggest that 6-week heavy load training induces the dysfunction of immunity and neuroendocrine responses, which might be one of the underlying mechanisms of immune dysfunction in overtraining. PMID- 12628446 TI - Inhibitory effects of total flavones of Hippophae Rhamnoides L on thrombosis in mouse femoral artery and in vitro platelet aggregation. AB - Total flavones of Hippophae Rhamnoides L (TFH) are extracted from Sea buckthorn, a Chinese herbal medicine. Sea buckthorn has antioxidant, anti-ulcerogenic and hepato-protective actions, and its berry oil is reported to suppress platelet aggregation. Though it is frequently used for patients with thrombosis, the likely mechanism(s) and effects of TFH on thrombogenesis remain unclear. Thus, we have investigated the effect in-vivo of TFH on thrombogenesis and in vitro on platelet aggregation, comparing them to those of aspirin. We measured thrombotic occlusion time in a mouse femoral artery thrombosis model by the photochemical reaction between intravenously injected rose bengal and green light irradiation. In vitro platelet aggregation in whole blood was measured by single platelet counting. Thrombotic occlusion time was 8.5 +/- 0.6 min in the control group. TFH at a dose of 300 micro g/kg, intravenously administered 15 min before the rose bengal injection, significantly prolonged it to 11.6 +/- 1.0 min (P < 0.05), a similar effect on in-vivo thrombogenesis to that of aspirin. TFH at a concentration of 3.0 micro g/ml significantly (P < 0.01) inhibited in vitro platelet aggregation induced by collagen (2 micro g/ml) in a concentration dependent manner, in contrast TFH did not affect aggregation induced by arachidonic acid (80 micro M) and ADP (0.3 micro M). The results of the present study, in which TFH prevented in-vivo thrombogenesis, probably due to inhibition of platelet aggregation, suggest a possible clinical approach for the prevention of thrombosis. PMID- 12628447 TI - Evaluation of hepatoprotective effect of Gentiana olivieri herbs on subacute administration and isolation of active principle. AB - Hepatoprotective effect of Gentiana olivieri Griseb. (Gentianaceae) flowering herbs on subacute administration were studied using in vivo models in rats. For the activity assessment on carbon tetrachloride-induced hepatic damage following biochemical parameters were evaluated; plasma and hepatic tissue malondialdehyde formation, and liver tissue glutathione level, as well as plasma transaminase enzyme levels (aspartate transferase and alanine transferase). Results of biochemical tests were also confirmed by histopathological examination. Through bioassay-guided fractionation procedures isoorientin, a known C-glycosylflavone, was isolated from the ethyl acetate fraction as the active antihepatotoxic constituent by silica gel column chromatography. Isoorientin exhibited significant hepatoprotective effect at 15 mg/kg b.w. dose. PMID- 12628448 TI - Binding profile of Artocarpus integrifolia agglutinin (Jacalin). AB - Artocarpus integrifolia agglutinin (Jacalin) from the seeds of jack fruits has attracted considerable attention for its diverse biological activities and has been recognized as a Galbeta1-->3GalNAc (T) specific lectin. In previous studies, the information of its binding was limited to the inhibition results of monosaccharides and several T related disaccharides, but its interaction with other carbohydrate structural units occurring in natural glycans has not been characterized. For this reason, the binding profile of this lectin was studied by enzyme linked lectinosorbent assay (ELLSA) with our glycan/ligand collection. Among glycoproteins (gps) tested for binding, high density of multi-Galbeta1- >3GalNAcalpha1--> (mT(alpha)) and GalNAcalpha1-->Ser/Thr (mTn) containing gps reacted most avidly with Jacalin. As inhibitors expressed as nanograms yielding 50% inhibition, these mT(alpha) and mTn containing glycans were about 7.1 x 10(3), 4.0 x 10(5), and 7.8 x 10(5) times more potent than monomeric T(alpha), GalNAc, and Gal. Of the sugars tested and expressed as nanomoles for 50% inhibition, Tn containing peptides, T(alpha), and the human P blood group active disaccharide (P(alpha), GalNAcbeta1-->3Galalpha1-->) were the best and about 283 times more active than Gal. We conclude that the most potent ligands for this lectin are mTn, mT, and possibly P(alpha) glycotopes, while GalNAcbeta1- >4Galbeta1-->, GalNAcalpha1-->3Gal, GalNAcalpha1-->3GalNAc, and Galalpha1-->3Gal determinants were poor inhibitors. Thus, the overall binding profile of Jacalin can be defined in decreasing order as high density of mTn, and mT(alpha) >>> simple Tn cluster > monomeric T(alpha) > monomeric P(alpha) > monomeric Tn > monomeric T > GalNAc > Gal > Methylalpha1-->Man z.Gt; Man and Glc (inactive). Our finding should aid in the selection of this lectin for biological applications. PMID- 12628450 TI - Introduction: the non-neuronal cholinergic system in humans. PMID- 12628451 TI - Role of the non-neuronal cholinergic system in the eye: a review. AB - Pharmacologically active preparations directed towards modulating muscarinic receptor activity in the eye have been used for over 2000 years when extracts from Atropa belladonna were first applied to enhance eye appearance through pupillary dilation. The first clinically active drugs targeting a specific eye disease were anticholinesterases (e.g. ecothiophate) applied as eye drops to treat glaucoma in the 1960's. However, cataract was soon detected as a relatively frequent side effect and such drugs are now only used to treat glaucoma as a last resort. As muscarinic agonists have been found to reduce intraocular pressure both by decreasing aqueous humour production (through Na,K-ATPase pump inhibition) and increasing outflow (by muscle contraction), it is likely that treatments will be developed that target specific muscarinic subtypes. Recently, it has been shown that the M1 receptor subtype predominates in the lens. It is therefore important that this subtype is not targeted in future ocular therapies so that the side-effect of cataract is avoided. Form-deprived myopia resulting from an increased axial length in the affected eye can be reduced by the application of atropine. This effect has been achieved both in a chick model system and in human clinical trials, and in the former system atropine has been shown to reduce the production of scleral extracellular proteins. Carbachol stimulates tear fluid production through the activation of muscarinic receptors. Interestingly, at least part of the stimulation occurs via epidermal growth factor (EGF) receptors and although the precise signalling mechanisms are not completely understood, it has been shown that calcium mobilisation plays a critical role in both muscarinic and EGF receptor activity. It should be noted that in the four examples described above, the cell types responsible for producing the physiological output are non-neuronal in origin. Therefore cholinergic receptor activation plays diverse roles in the eye and pharmacological intervention based on specific receptor sub-types has potential benefit in a number of ocular problems. However, potential side effects have also recently been identified. PMID- 12628452 TI - Regulation of cholinergic gene expression by the neuron restrictive silencer factor/repressor element-1 silencing transcription factor. AB - The role of protein kinase A in regulating transcription of the cholinergic gene locus, which contains both the vesicular acetylcholine transporter gene and the choline acetyltransferase gene, was investigated in PC12 cells and a protein kinase A deficient PC12 mutant, A126.1B2 in which transcription of the locus is reduced. The site of action of protein kinase A was localized to a neuron restrictive silencer element/repressor element-1 (NRSE/RE-1) within the upstream region of the cholinergic gene locus. The neuron restrictive silencer factor/repressor element-1 silencing transcription factor (NRSF/REST), the transcription factor which binds to NRSE/RE-1, was expressed at similar levels in both PC12 and A126.1B2. Although nuclear extracts containing NRSF/REST from A126.1B2 exhibited binding to NRSE/RE-1, nuclear extracts from PC12 cells did not. The NRSF/REST isoform repressor element-1 silencing transcription factor-4 (REST4) was found to be expressed in PC12 cells, but not in the protein kinase A deficient PC12 cell line. REST4 inhibited the binding of NRSF/REST to NRSE/RE-1 as determined by gel mobility shift assays. Co-immunoprecipitation was used to demonstrate interaction between NRSF/REST and REST4. Expression of recombinant REST4 in the protein kinase A deficient PC12 cell line was sufficient to transcriptionally activate the cholinergic gene locus. Thus in PC12 cells protein kinase A promotes the production of REST4, which in turn de-represses of the cholinergic gene locus by inactivating the transcription repressor NRSF/REST. PMID- 12628454 TI - The expression and biological role of the non-neuronal cholinergic system in the ovary. AB - Functioning of the ovary depends on an interplay between hormones, locally produced growth factors and neurotransmitters. Neurotransmitters are delivered to the ovary via its sympathetic innervation and originate from intrinsic nerve cells expressing catecholaminergic and peptidergic traits. We found that the nerve fibers and nerve cells of the ovary were however not immunoreactive for the ACh-synthesizing enzyme, choline-acetyl transferase (ChAT). Immunoreactivity was instead detected in ovarian endocrine cells, namely granulosa cells (GCs), of rodents and primates. Importantly, isolated GCs produce ACh. Thus, endocrine cells are an unexpected non-neuronal source of ACh in the ovary. GCs in vivo and in vitro also contain ACh-receptors of the muscarinic subtype (MR), namely M1 and M5. MR of human GCs are functional and linked to rapid increases in intracellular calcium levels. A role of ovarian ACh/MR in the crucial process of cell proliferation is suggested by the observation that in growing follicles, ChAT immunoreactive GCs co-express "proliferating cell nuclear antigen" (PCNA) and that cholinergic agents stimulate cell proliferation of human GCs in vitro. This proliferative effect is associated with rapid disruption of gap junction communication and phosphorylation of connexin 43. In addition, calcium-dependent channels are activated. Ongoing studies have begun to identify down-stream effects of M1/5 activation in GCs, which include, for example, expression of a transcription factor (egr-1). In summary, ovarian endocrine cells are sources and targets of ACh. We propose that an as yet unexplored intraovarian cholinergic system exists, which contributes to physiological ovarian tissue remodeling by stimulation of cell proliferation. PMID- 12628453 TI - Expression of the acetylcholine release mechanism in various cells and reconstruction of the release mechanism in non-releasing cells. AB - After loading cells in culture with acetylcholine (ACh), it was possible to identify cells that express a calcium-dependent release mechanism and cells that do not release. Mediatophore transfection restored the release capability of non releasing cells. The transfection of choline acetyltransferase and the vesicular ACh transporter (VAChT) in cells that have already mediatophore in their membrane enables to study the effect of VAChT on the release kinetics. We also studied the properties of the mediatophore "pore" as a function of the concentration of ACh and also its temporal properties. A reconstruction of the release mechanism in cells particularly graftable cells, appears now possibly for ACh and probably for other transmitters. PMID- 12628455 TI - Muscarinic receptor subtypes mediating central and peripheral antinociception studied with muscarinic receptor knockout mice: a review. AB - To gain new insight into the physiological and pathophysiological roles of the muscarinic cholinergic system, we generated mutant mouse strains deficient in each of the five muscarinic acetylcholine receptor subtypes (M(1)-M(5)). In this chapter, we review a set of recent studies dealing with the identification of the muscarinic receptor subtypes mediating muscarinic agonist-dependent analgesic effects by central and peripheral mechanisms. Most of these studies were carried out with mutant mouse strains lacking M(2) or/and M(4) muscarinic receptors. It is well known that administration of centrally active muscarinic agonists induces pronounced analgesic effects. To identify the muscarinic receptors mediating this activity, wild-type and muscarinic receptor mutant mice were injected with the non-subtype-selective muscarinic agonist, oxotremorine (s.c., i.t., and i.c.v.), and analgesic effects were assessed in the tail-flick and hot-plate tests. These studies showed that M(2) receptors play a key role in mediating the analgesic effects of oxotremorine, both at the spinal and supraspinal level. However, studies with M(2)/M(4) receptor double KO mice indicated that M(4) receptors also contribute to this activity. Recent evidence suggests that activation of muscarinic receptors located in the skin can reduce the sensitivity of peripheral nociceptors. Electrophysiological and neurochemical studies with skin preparations from muscarinic receptor mutant mice indicated that muscarine induced peripheral antinociception is mediated by M(2) receptors. Since acetylcholine is synthesized and released by different cell types of the skin, it is possible that non-neuronally released acetylcholine plays a role in modulating peripheral nociception. Our results highlight the usefulness of muscarinic receptor mutant mice to shed light on the functional roles of acetylcholine released from both neuronal and non-neuronal cells. PMID- 12628456 TI - The non-neuronal cholinergic system in humans: expression, function and pathophysiology. AB - Acetylcholine, a prime example of a neurotransmitter, has been detected in bacteria, algae, protozoa, and primitive plants, indicating an extremely early appearance in the evolutionary process (about 3 billion years). In humans, acetylcholine and/or the synthesizing enzyme, choline acetyltransferase (ChAT), have been found in epithelial cells (airways, alimentary tract, urogenital tract, epidermis), mesothelial (pleura, pericardium), endothelial, muscle and immune cells (mononuclear cells, granulocytes, alveolar macrophages, mast cells). The widespread expression of non-neuronal acetylcholine is accompanied by the ubiquitous presence of cholinesterase and receptors (nicotinic, muscarinic). Thus, the non-neuronal cholinergic system and non-neuronal acetylcholine, acting as a local cellular signaling molecule, has to be discriminated from the neuronal cholinergic system and neuronal acetylcholine, acting as neurotransmitter. In the human placenta anti-ChAT immunoreactivity is found in multiple subcellular compartments like the cell membrane (microvilli, coated pits), endosomes, cytoskeleton, mitochondria and in the cell nucleus. These locations correspond with the results of experiments where possible functions of non-neuronal acetylcholine have been identified (proliferation, differentiation, organization of the cytoskeleton and the cell-cell contact, locomotion, migration, ciliary activity, immune functions). In the human placenta acetylcholine release is mediated by organic cation transporters. Thus, structural and functional differences are evident between the non-neuronal and neuronal cholinergic system. Enhanced levels of acetylcholine are detected in inflammatory diseases. In conclusion, it is time to revise the role of acetylcholine in humans. Its biological and pathobiological roles have to be elucidated in more detail and possibly, new therapeutical targets may become available. PMID- 12628457 TI - Functional role of alpha7 nicotinic receptor in physiological control of cutaneous homeostasis. AB - Non-neuronal nicotinic acetylcholine receptors (nAChRs) are abundantly expressed in skin and their function remains to be elucidated. Herein, we report that cutaneous alpha7 nAChR plays a role in the physiological control of cutaneous homeostasis. We studied in vitro effects of functional inactivation of alpha7 receptor on the expression of apoptosis regulators in keratinocytes (KC) lacking alpha7 nAChR, and extracellular matrix regulators in the skin of alpha7 knockout (KO) mice. Elimination of the alpha7 component of nicotinergic signaling in KC decreased relative amounts of the pro-apoptotic Bad and Bax at both the mRNA and the protein levels, suggesting that alpha7 nAChR is coupled to stimulation of keratinocyte apoptosis. The skin of alpha7 KO mice featured decreased amounts of the extracellular matrix proteins collagen 1alpha1 and elastin as well as the metalloproteinase-1. Taken together, these results suggest an important role for alpha7 nAChR in mediating plethoric effects of non-neuronal acetylcholine on cutaneous homeostasis. PMID- 12628458 TI - The M4 muscarinic receptor-selective effects on keratinocyte crawling locomotion. AB - We have investigated how the cholinergic system of epidermal keratinocytes (KC) controls migratory function of these cells. Several molecular subtypes of muscarinic acetylcholine receptors (mAChRs) have been detected in KC. Early results suggested that M(4) is the predominant mAChR regulating cell motility. To determine muscarinic effects on lateral migration of KC, we used an agarose gel keratinocyte outgrowth system (AGKOS) which provides for measurements of the response of large cell populations (> 10(4) cells). Muscarine produced a dose dependent stimulatory effect on cell migration (p < 0.05). This activity was abolished by atropine, which decreased migration distance when given alone. To identify the mAChR subtype(s) mediating these muscarinic effects, we substituted atropine with subtype-selective antagonists. Tropicamide (M(4)-selective) was more effective at decreasing the migration distance than pirenzepine and 4-DAMP at nanomolar concentrations. We then compared lateral migration of KC obtained from M(4) mAChR knockout mice with that of wild-type murine KC, using AGKOS. In the absence of M(4) mAChR, the migration distance of KC was significantly (p < 0.05) decreased. These results indicate that the M(4) mAChR plays a central role in mediating cholinergic control of keratinocyte migration by endogenous acetylcholine produced by these cells. PMID- 12628459 TI - Proliferative effect of acetylcholine on rat trachea epithelial cells is mediated by nicotinic receptors and muscarinic receptors of the M1-subtype. AB - Acetylcholine (ACh), synthesized in mammalian non-neuronal cells such as epithelial cells of the airways, digestive tract and skin, is involved in the regulation of basic cell functions (so-called non-neuronal cholinergic system). In the present experiments rat trachea epithelial cells have been cultured to study the proliferative effect of applied ACh by [3H]thymidine incorporation. ACh (exposure time 24 h) caused a concentration-dependent increase in cell proliferation with a doubling of the [3H]thymidine incorporation at a concentration of 0.1 microM. This effect was partly reduced by 30 microM tubocurarine and completely abolished by the additional application of 1 microM atropine. The stimulatory effect of acetylcholine, remaining in the presence of tubocurarine, was prevented by 1 microM pirenzepine (preferentially acting at M1 receptors), but neither by 1 microM AFDX 116 (preferentially acting at M2 receptors) nor by 1 microM hexahydrosiladifenidol (preferentially acting at M3 receptors). The combination of tubocurarine and pirenzepine halved the basal [3H]thymidine incorporation. In conclusion, ACh produces a proliferative effect in rat trachea epithelial cells, the effect being mediated by both nicotinic receptors and muscarinic receptors of the M1-subtype. PMID- 12628461 TI - Expression of the high-affinity choline transporter CHT1 in epithelia. AB - Uptake of choline by the high-affinity choline transporter CHT1 is the rate limiting step in neuronal acetylcholine (ACh) synthesis. Here, we investigated by RT-PCR, in-situ hybridisation, immunohistochemistry, and Western blotting whether CHT1 is also expressed in cholinergic epithelia. CHT1-mRNA and -protein were detected in keratinocytes of human skin, rat skin and tongue, the human keratinocyte cell line HaCaT, and the ciliated cells of the rat tracheal epithelium. Immunohistochemically, CHT1 was predominantly localized to the epithelial cell membranes, in case of ciliated tracheal cells it was restricted to the apical membrane. This is the first study to demonstrate the expression of CHT1 in non-neuronal cells. The close apposition of CHT1 to reported sites of localization of choline acetyltransferase in these cells is strongly in favour of ACh synthesis being fuelled by choline uptake via CHT1 in these epithelia. PMID- 12628460 TI - Keratinocyte acetylcholine receptors regulate cell adhesion. AB - We investigated the mechanism mediating cholinergic control of cell-to-cell adhesion of human epidermal keratinocytes (KC) by non-neuronal acetylcholine produced by KC themselves. We first measured cholinergic effects on the expression of desmoglein (Dsg) 1 and 3 in KC using the semi-quantitative immunofluorescence and Western blot assays. Monolayers of KC were treated overnight with 0.25 mM of the cholinergic agonist carbachol (CCh) or the acetylcholinesterase inhibitor pyridostigmine bromide (PBr). Both CCh and PBr increased the relative amounts of Dsg 1 and Dsg 3. To determine the role for cholinergic receptor-mediated phosphorylation of Dsg molecules in assembly/disassembly of keratinocyte desmosomes, we tested the effects of a cholinergic antagonist on keratinocyte adhesion and Dsg phosphorylation status in DJM-1 cell line. Atropine (Atr), 0.02 mM, induced rapid detachment of cells from each other (acantholysis), and also increased phosphorylation of Dsg 3 by 33%. The Atr-dependent phosphorylation of Dsg 3 was inhibited in the presence of 0.5 mM CCh. Thus, keratinocyte cholinergic receptors regulate desmosomal adhesion of KC by altering the level of expression of both Dsg 1 and Dsg 3 and the phosphorylation status of Dsg 3. PMID- 12628462 TI - Nicotinic acetylcholine receptor subunits and receptor activity in the epithelial cell line HT29. AB - In the present study we have used RT-PCR to investigate nicotinic acetylcholine receptor (nAChR) subunit expression, and studied the effect of nicotine on TNFalpha-induced cytokine (IL-8) release in the epithelial cell line HT29. RNA was extracted using a commercial kit and amplified by RT-PCR. RT-PCR products were separated by electrophoresis and visualised using ethidium bromide. IL-8 release was measured by ELISA from cells activated for 6 h with TNFalpha (50 ng ml(-1)) in the absence and presence of nicotine (10(-11)-10(-6) M). HT29 cells contained mRNA for beta1, alpha4, alpha5, and alpha7 nAChR subunits. Activation of HT29 cells increased IL-8 release from undetectable amounts to 3.92 +/- 0.51 ng ml(-1) (n = 5). Nicotine significantly inhibited TNFalpha-induced IL-8 release in a concentration related manner with peak inhibition occurring at 10(-7) M (2.39 +/- 0.78 ng ml(-1), n = 5). Our data suggests that, while HT29 cells express mRNA for nAChR subunits, the only nAChR subunits that could form functional receptors and inhibit IL-8 release are alpha7. PMID- 12628463 TI - Rat arteries contain multiple nicotinic acetylcholine receptor alpha-subunits. AB - We investigated the occurrence and distribution of the ligand-binding alpha subunits of nicotinic acetylcholine receptors in the rat arterial system in situ by means of RT-PCR and immunohistochemistry. Except the alpha9-subunit, all other mammalian non-muscular alpha-subunits were expressed in the arterial wall--either in endothelial or in smooth muscle cells--suggesting it as a direct target of nicotine and endogenous acetylcholine. The distribution pattern of alpha-subunits found in smooth muscle cells varied considerably among the individual elastic, muscular and intraparenchymal arteries investigated, suggesting that non-neuronal cholinergic signalling via nicotinic receptors in the vascular wall includes components that are highly specific for individual arteries. PMID- 12628464 TI - The lymphocytic cholinergic system and its biological function. AB - Lymphocytes are now known to possess the essential components for a non-neuronal cholinergic system. These include acetylcholine (ACh); choline acetyltransferase (ChAT), its synthesizing enzyme; and both muscarinic and nicotinic ACh receptors (mAChRs and nAChRs, respectively). Stimulating lymphocytes with phytohemagglutinin, a T-cell activator; Staphylococcus aureus Cowan I, a B-cell activator; or cell surface molecules enhances the synthesis and release of ACh and up-regulates expression of ChAT and M(5) mAChR mRNAs. Activation of mAChRs and nAChRs on lymphocytes elicits increases in the intracellular Ca(2+) concentration and stimulates c-fos gene expression and nitric oxide synthesis. On the other hand, long-term exposure to nicotine down-regulates expression of nAChR mRNA. Abnormalities in the lymphocytic cholinergic system have been detected in spontaneously hypertensive rats and MRL-lpr mice, two animal models of immune disorders. Taken together, these data present a compelling picture in which immune function is, at least in part, under the control of an independent non neuronal lymphocytic cholinergic system. PMID- 12628465 TI - Expression and function of the non-neuronal cholinergic system in endothelial cells. AB - Increasing evidence has shown the expression of the non-neuronal cholinergic system in endothelial cells. In the present experiments the expression of choline acetyltransferase (ChAT) was investigated in human endothelial cells by anti-ChAT immunohistochemistry and anti-ChAT immunofluorescence. Positive ChAT immunoreactivity was found in cultures of human umbilical endothelial cells (HUVEC) and a human angiosarcoma cell line (HAEND). In HUVEC and HAEND choline acetyltransferase activity and small amounts of acetylcholine were also detected. Positive ChAT-immunoreactivity was demonstrated in situ in endothelial cells of the human umbilical cord. In addition, in experiments with confocal laser scanning microscopy positive anti-ChAT immunoreactivity was found in situ in endothelial cells of human skin blood vessels. In the first functional experiments with HUVEC acetylcholine appeared to mediate a small facilitatory effect on the expression of intracellular adhesion molecule-1. The present experiments demonstrate the wide existence of ChAT in human endothelial cells. Further experiments are addressed to elucidate the biological role of acetylcholine in the endothelium and possible differences between the different subtypes of endothelial cells. PMID- 12628466 TI - A beta-induced TNF-alpha expression and acetylcholine action in mouse glial cells. AB - The brains in patients with Alzheimer's disease show chronic inflammatory responses characterized by activated glial cells and increased expression of cytokines. It is of interest to determine whether acetylcholine (ACh) affects Abeta-induced cytokine expression in the glial cells. Since it has been shown that alpha7 subunits of nicotinic ACh receptors are expressed in glial cells and that Abeta(1-42) binds to alpha7, we examined the effects of cholinergic agonists, carbachol, nicotine and oxotremorine-M, on Abeta-induced TNF-alpha expression in mouse glial cells. We did not observe any regulatory effects of ACh on Abeta-induced TNF-alpha transcription in the glial cells. We discuss the pathophysiological roles of ACh in glial cells in the brains of patients with Alzheimer's disease. PMID- 12628467 TI - The presence and functions of muscarinic receptors in human T cells: the involvement in IL-2 and IL-2 receptor system. AB - The existence and functions of muscarinic acetylcholine (mACh) receptors in human T lymphocytes were investigated. RT-PCR analysis demonstrated the presence of M(1) and M(2) subtypes of mACh receptors in human T lymphocytes. Pretreatment with oxotremorine-M (Oxo-M) caused the increase in phytohemagglutinin-induced IL 2 production. Since 4-DAMP suppressed Oxo-M-caused enhancement in IL-2 production, M(1) receptors seem to be involved in the enhancement of the production. Oxo-M stimulated IL-2 receptor mRNA expression and DNA synthesis. Our results suggest that muscarinic receptors, perhaps M(1) receptors are involved in the enhancement of TCR-induced IL-2 production and IL-2 receptor expression in human T lymphocytes. Thus muscarinic receptors positively modulate cell growth in human T lymphocytes by the autocrine mechanism through enhancing expression of both IL-2 and the receptors. PMID- 12628468 TI - Expression of multiple mRNA species for choline acetyltransferase in human T lymphocytes. AB - Acetylcholine (ACh) is synthesized by choline acetyltransferase (ChAT) in cholinergic neurons. However, both ACh and mRNA for ChAT are expressed in mononuclear leukocytes and various human leukemic T-cell lines. Multiple ChAT mRNA species (R-, N0-, N1-, N2-, and M-types) having an identical coding region and different 5'-noncoding regions have been discovered in human brain and spinal cord. These mRNAs are transcribed by a combination of use of different promoter regions and alternative splicing. However, which types of ChAT mRNA species are expressed in T-lymphocytes remains to be elucidated. In the present study, we used two human leukemic T-cell lines, CCRF-CEM (CEM) and MOLT-3, which express the same ChAT mRNA as that in the nervous system. Major mRNA species in CEM were N2- and M-type, and to a lesser extent N1-type, while MOLT-3 expressed only N2 type. Neither CEM nor MOLT-3 expressed R-type mRNA. We previously found a lack of mRNA expression encoding vesicular acetylcholine transporter (VAChT) in CEM and MOLT-3, which mediates ACh transport to synaptic vesicles in cholinergic neurons. These findings suggest that the mechanisms regulating ChAT mRNA expression in T lymphocytes differ from those in cholinergic neurons. PMID- 12628469 TI - Detection of the high-affinity choline transporter in the MOLT-3 human leukemic T cell line. AB - We previously showed that lymphocytes possess the necessary components to constitute an independent, non-neuronal cholinergic system; these include acetylcholine (ACh) itself, choline acetyltransferase (the ACh-synthesizing enzyme), and both muscarinic and nicotinic ACh receptors (AChRs). In addition, we showed that stimulation of AChRs with their respective agonists elicits a variety of biochemical and functional effects, suggesting that lymphocytic cholinergic system is involved in the regulation of immune function. In nerve terminals, choline taken up via the high-affinity choline transporter (CHT1) is exclusively utilized for ACh synthesis. In the present study, therefore, we investigated the expression of CHT1 in T-lymphocytes. Reverse transcription-polymerase chain reaction analysis revealed that MOLT-3 cells, a human leukemic T-cell line used as a T-lymphocyte model, expressed CHT1 mRNA, but that the CEM and Jurkat T-cell lines did not. Consistent with that finding, specific binding of [3H]hemicholinium-3 (HC-3), an inhibitor of CHT1, and HC-3-sensitive [3H]choline uptake were also detected in MOLT-3 cells. These results suggest that CHT1 plays a role in mediating choline uptake in T-lymphocytes and provides further evidence for the presence of an independent lymphocytic cholinergic system. PMID- 12628470 TI - Mucocutaneous cholinergic system is targeted in mustard-induced vesication. AB - The purpose of this research is to elucidate the pharmacological mechanism mediating vesicating effects of sulfur mustard (HD) and identify an antidote to its action. HD causes blisters because epithelial cells lose their attachments. Epithelial cell adhesion is under control of the local cytotransmitter acetylcholine (ACh) working through the muscarinic and nicotinic receptor, mAChR and nAChR, classes expressed by epithelial cells. In this study, nitrogen mustard (NM)-a structural analog of HD-was used to elucidate the mechanism of vesicating effects of mustards in mucocutaneous tissues. NM caused cell detachment and cholinergic agents antagonized its effect. Radioligand binding inhibition experiments showed that NM binds to the ligand-binding site of ACh receptors (AChRs) of both classes. Ligation of AChRs on the cell membrane of keratinocytes (KC) and bronchial epithelial cells (BEC) with NM increased total esterolytic activity of serine proteinases (TEASP). Antagonists of both classes of AChRs, atropine and mecamylamine, diminished NM-induced changes, suggesting that the pathobiological effects of NM on KC and BEC result from an agonist-like degradation of ligated AChRs, predominantly of the muscarinic class. Thus, biological effects of NM on cell adhesion were antagonist-like, whereas its pharmacological effect on TEASP was agonist-like. These findings support a hypothesis that pharmacologic protection from the vesicating action of HD can be achieved by using cholinergic drugs. PMID- 12628471 TI - Acetylcholine increase in amniotic fluid of experimental rats for intrauterine growth retardation. AB - Previous reports from this laboratory have demonstrated evidence for synthesis and release of acetylcholine (ACh) and catecholamines (CAs) by human amniotic epithelial cells (HAEC) and the presence of ACh and CAs in amniotic fluid. To study the physiological role of amniotic ACh, we used an experimental pregnant rat model for intrauterine growth retardation. Prior to this experiment, we confirmed the presence of choline acetyltransferase in the HAEC by immunocytochemical staining. Amniotic fluid was collected at 48 and 72 h after a transient ligation of the uterine vessels near the lower and upper ends of the right horn of the pregnant rat. The ACh concentration in the amniotic fluid from rats received intrauterine ischemia increased with time to a greater degree compared with the control rat, although the increase was not statistically significant. These results suggest that intrauterine hypoxic conditions cause a tendency to increase ACh concentrations in the amniotic fluid. PMID- 12628472 TI - Nitric oxide (NO) synthase mRNA expression and NO production via muscarinic acetylcholine receptor-mediated pathways in the CEM, human leukemic T-cell line. AB - Nitric oxide (NO) is synthesized from L-arginine by neuronal, endothelial and inducible isoforms of NO synthase (nNOS, eNOS and iNOS, respectively) and is involved in the regulation of a variety of physiological functions, including immune activity. In vascular endothelial cells, stimulation of M(3) subtype of muscarinic acetylcholine receptors (mAChRs) triggers NO synthesis by eNOS. Human lymphocytes express several mAChR subtypes and their stimulation increases the intracellular free Ca(2+) concentration and up-regulates c-fos gene expression. While the above findings suggest involvement of the lymphocytic cholinergic system in the regulation of immune function, little is known on NOS expression and NO synthesis in T-lymphocytes. In the present study, using reverse transcription-polymerase chain reaction, we found that CEM cells express mRNAs encoding iNOS and nNOS, but not for eNOS. In addition, using quantitative fluorescence microscopy and a novel NO-sensitive fluorescent indicator, DAF-2, we found that oxotremorine-M (Oxo-M) (100 microM), a non-selective mAChR agonist, enhances NO production in the cells. This effect of Oxo-M was antagonized by pirenzepine (10 microM), an antagonist acting preferentially at M(1) mAChR and by atropine (10 microM). Also 4-DAMP (10 microM), an antagonist acting preferentially at M(3) mAChR, reduced significantly the effect of Oxo-M, while AFDX-116 (10 microM), an antagonist acting preferentially at M(2) mAChR, was ineffective. These findings suggest that T-lymphocytes express functional mAChRs linked to NO synthesis by nNOS and/or iNOS. PMID- 12628473 TI - Nicotine-induced Ca2+ signaling and down-regulation of nicotinic acetylcholine receptor subunit expression in the CEM human leukemic T-cell line. AB - We previously showed that T- and B-lymphocytes express both muscarinic and nicotinic acetylcholine (ACh) receptors (mAChR and nAChR, respectively), and that stimulation of M(3) mAChRs on lymphocytes increases the intracellular free Ca(2+) concentration ([Ca(2+)](i)) and up-regulates c-fos gene expression. Little is known about the effects of nicotinic stimulation on lymphocyte function, however. We therefore investigated the acute effect of nicotine on [Ca(2+)](i) in CEM cells, a model of T-lymphocytes, using confocal laser scanning microscopy with fluo-3, a Ca(2+)-sensitive fluorescent indicator. In addition, we examined the long-term effect of nicotine on the expression of selected nAChR subunits using semiquantitative reverse transcription-polymerase chain reaction analysis. In the presence of extracellular Ca(2+), nicotine (30 microM) evoked rapid, transient increases in [Ca(2+)](i). This effect was concentration-dependently inhibited by the alpha7 nAChR subunit antagonists, alpha-bungarotoxin (0.01-10 microM) and methyllycaconitine (0.01-10 mM), suggesting that the alpha7 nAChR subunit mediates Ca(2+) signaling in T-lymphocytes. Nicotine (0.01-10 microM) also concentration-dependently down-regulated expression of mRNAs for all the nAChR subunits tested: expression of the alpha6 and alpha7 subunits was down-regulated within 1 week, while expression of the alpha3 and alpha5 subunits declined gradually throughout the 8-week experimental period. These findings indicate that nicotine--and therefore likely smoking--affects immune function by suppressing expression of the neuronal nAChR subtype involved in Ca(2+) signaling in lymphocytes. PMID- 12628474 TI - Synthesis of acetylcholine by lung cancer. AB - The role of autocrine growth factors in the stimulation of lung cancer growth is well established. Nicotine is an agonist for acetylcholine receptors and stimulates lung cancer growth. This suggests that if lung cancers synthesize acetylcholine (ACh), then ACh may be an autocrine growth factor for lung cancer. Analysis of normal lung demonstrated that the cells of origin of lung cancers express the proteins necessary for non-neuronal ACh storage and synthesis. Analysis of mRNA from squamous cell lung carcinoma, small cell lung carcinoma (SCLC) and adenocarcinoma showed synthesis of choline acetyltransferase (ChAT) and nicotinic receptors. Immunohistochemical analysis of a retrospective series of SCLC and adenocarcinomas showed that more than 50% of the lung cancers screened expressed ChAT and nicotinic receptors. To study the effect of endogenous ACh synthesis on growth, SCLC cell lines were studied. SCLC cell lines were found to express ChAT mRNA and to secrete ACh into the medium as measured by HPLC separation and enzymatically-coupled electrochemical detection. The SCLC cell line NCI-H82 synthesized highest levels of ACh. Showing that the endogenously synthesized ACh interacted with its receptors to stimulate cell growth, addition of muscarinic and nicotinic antagonists slowed H82 cell proliferation. These findings demonstrate that lung cancer cell lines synthesize and secrete ACh to act as an autocrine growth factor. The existence of a cholinergic autocrine loop in lung cancer provides a basis for understanding the effects of nicotine in cigarette smoke on lung cancer growth and provides a new pathway to investigate for potential therapeutic approaches to lung cancer. PMID- 12628475 TI - Increased acetylcholine levels in skin biopsies of patients with atopic dermatitis. AB - Recent experimental evidence indicates that non-neuronal acetylcholine is involved in the regulation of basic cell functions. Here we investigated the cholinergic system in the skin of healthy volunteers and patients with atopic dermatitis (AD). The synthesizing enzyme, choline-acetyltransferase (ChAT), was studied by anti-ChAT immunohistochemistry and enzyme assay. Skin biopsies taken from healthy volunteers and from AD patients were separated into the 2 mm superfical (epidermis and upper dermis) and 3 mm underlying portion (deeper dermis and subcutis). ChAT enzyme activity was detected in homogenized skin and subcutaneous fat (about 13 nmol/mg protein/h). ChAT immunoreactivity was expressed in keratinocytes, hair papilla, sebaceous and eccrine sweat glands, endothelial cells and mast cells. In healthy volunteers the superficial and underlying portion of skin biopsies contained 130 +/- 30 and 550 +/- 170 pmol/g acetylcholine (n = 12), respectively. In AD patients (n = 7) acetylcholine was increased 14-fold in the superficial and 3-fold in the underlying biopsy portion. The present study demonstrates the widespread expression of ChAT protein in the vast majority of human skin cells. Tissue levels of acetylcholine are greatly (14 fold) enhanced in the superficial 2 mm skin of AD patients. PMID- 12628476 TI - Muscarinic signaling in carcinoma cells. AB - We previously reported that activation of M(3) muscarinic acetylcholine receptors (mAChR) generates anti-proliferative signals and stimulates cadherin-mediated adhesion in the SCC-9 small cell lung carcinoma (SCLC) cell line. The current study was undertaken to determine the frequency of functional mAChR expression among different SCLC cell lines, and to test the ability of mAChR to generate anti-proliferative signals in different SCLC cell lines. The potential role of Rac1 in SCLC cell-cell adhesion was also investigated. Exposure to the mAChR agonist carbachol induces robust Ca(2+) mobilization (indicated by intracellular fluorescence of the Ca(2+)-binding dye Indo-1) in three SCLC cell lines (SCC-9, SCC-15, and NCI-H146), modest Ca(2+) mobilization in one SCLC cell line (NCI H209), and no detectable Ca(2+) mobilization in two SCLC cell lines (SCC-18 and NCI-H82). The M(3) mAChR-selective antagonist 4-diphenylacetoxy-N methylpiperidine methiodide inhibits Ca(2+) mobilization in all SCLC cell lines responding to carbachol. Incubation with carbachol for four hours significantly inhibits [3H]thymidine uptake in three of the four SCLC cell lines expressing functional mAChR (SCC-9, SCC-15, and NCI-H146 cells), but does not significantly alter [3H]thymidine uptake in the other SCLC cell lines examined. These results indicate that SCLC cell lines often express functional mAChR which elicit anti proliferative signals when activated. To investigate the role of Rac1 in SCLC adhesion, SCC-9 cells were transiently transfected with cDNA constructs coding for Rac1, constitutively active Rac1(Val-12), or dominant negative Rac1(Asn-17) tagged to green fluorescent protein (GFP). SCC-9 cells expressing GFP-tagged constitutively active Rac1(Val-12) exhibit increased cell-cell adhesion in comparison to cells expressing GFP-Rac1 or GFP-Rac1(Asn-17). Constitutively active GFP-Rac1(Val-12), but not GFP-Rac1 or GFP-Rac1(Asn-17), accumulates at cell-cell junctions in SCC-9 cells. These results indicate that activated Rac1 increases SCLC cell-cell adhesion, consistent with the possibility that Rac1 activation contributes to increased SCLC cell-cell adhesion induced by mAChR stimulation. These findings indicate that activation of mAChR may play a significant role in regulating the proliferation and adhesion of SCLC cells. The demonstration by other investigators that acetylcholine is expressed by a variety of cells in the airways supports the possibility that acetylcholine may activate mAChR expressed by SCLC cells in primary tumors. PMID- 12628477 TI - Differential effects of YM440 a hypoglycemic agent on binding to a peroxisome proliferator-activated receptor gamma and its transactivation. AB - Peroxisome proliferator-activated receptor (PPAR) gamma is a ligand-inducible transcription factor mediating glucose and lipid metabolism. Prior studies showed that YM440 ameliorated hyperglycemia in diabetic mice without affecting body fat weight or PPARgamma transactivation. In this study we have examined further the effects of YM440 on PPARgamma binding, transactivation and conformational change. YM440, pioglitazone and rosiglitazone displaced [3H]rosiglitazone from PPARgamma with K(i) values of 4.0, 3.1, and 0.20 microM, indicating that YM440 was comparable to pioglitazone and 20-fold less potent than rosiglitazone. Although pioglitazone and rosiglitazone increased both PPARgamma transactivation in cells expressing human full-length PPARgamma2 or GAL4-PPARgamma and mRNA expression of PPARgamma responsive genes in 3T3-L1 cells, YM440 had weak effects on PPARgamma transactivation and mRNA expression being 550- to 790-fold and 36- to 110-fold less active than rosiglitazone, respectively. YM440 and rosiglitazone induced interaction between PPARgamma and the transcriptional cofactor, p300 or SRC-1, but YM440 was 151- and 1091-fold less potent than rosiglitazone, respectively. The weak transcriptional activity of YM440 was not due to poor cell permeability. Limited trypsin digestion of the full-length human PPARgamma2 with YM440 or rosiglitazone showed distinct patterns of digestion, suggesting a difference in the conformational change of PPARgamma. When db/db mice were treated with YM440 (100mg/kg) for 28 days, YM440 increased hepatic glucokinase expression but not adipose tissue FABP and UCP1 expression, indicating a tissue selective expression of PPARgamma-related genes. Unique properties regarding the binding transactivation of PPARgamma by YM440 may lead to the hypoglycemic activity without affecting body fat weight in diabetic mice. PMID- 12628478 TI - Cellular transformation of the investigational new anticancer drug NB1011, a phosphoramidate of 5-(2-bromovinyl)-2'-deoxyuridine, results in modification of cellular proteins not DNA. AB - NB1011 [E-5-(2-bromovinyl)-2'-deoxyuridine-5'-(L-methylalaninyl) phenylphosphoramidate], a phosphoramidate prodrug of E-5-(2-bromovinyl)-2' deoxyuridine-5'-monophosphate (BVdUMP), is an investigational new anticancer drug. NB1011 targets thymidylate synthase (TS), which catalyzes the transformation of BVdUMP into cytotoxic reaction products. Due to the elevated levels of TS expression in tumor cells compared to normal cells, these cytotoxic products are preferentially generated inside tumor cells, and, as expected, NB1011 is more toxic to cells with higher levels of TS expression. Therefore, NB1011 therapy should kill tumor cells without severely damaging normal cells. Radiolabeled NB1011 was used to determine the intracellular fate of NB1011 reaction products and, possibly, the mechanism of action of this investigational new drug. We found significant incorporation of the radiolabel into cellular macromolecules. In contrast to our expectations that NB1011 product(s) would be incorporated into DNA, we discovered that cellular proteins were the labeled macromolecular fraction. Herein, we report that the intracellular transformation of NB1011 involves formation of the corresponding monophosphate, TS-dependent transformation into highly reactive intermediates, and subsequent incorporation into cellular proteins. TS itself appears to escape irreversible inactivation. Our data suggest that protein modification not DNA incorporation accounts for the therapeutic effect of NB1011. The proposed mechanism is rather unexpected for a nucleotide analogue and could lead to the discovery of new cellular protein targets for future drug design. PMID- 12628479 TI - Statin effects on cholesterol micro-domains in brain plasma membranes. AB - Recent epidemiological studies revealed inhibitors of the hydroxymethylglutaryl coenzyme A reductase, so-called statins, to be effective in lowering the prevalence of Alzheimer's disease (AD). In vitro, statins strongly reduced the cellular amyloid beta-protein load by modulating the processing of the amyloid beta precursor protein. Both observations are probably linked to cellular cholesterol homeostasis in brain. So far, little is known about brain effects of statins. Recently, we could demonstrate that treatment of mice with the lipophilic compound lovastatin resulted in a discrete reduction of brain membrane cholesterol levels. To follow up these findings, we subsequently carried out a further in vivo study including lovastatin and simvastatin as lipophilic agents, as well as pravastatin as a hydrophilic compound, focussing on their efficiency to affect subcellular membrane cholesterol pools in synaptosomal plasma membranes of mice. In contrast to the hydrophilic pravastatin, the lipophilic lovastatin and simvastatin strongly reduced the levels of free cholesterol in SPM. Interestingly, lovastatin and pravastatin but not simvastatin significantly reduced cholesterol levels in the exofacial membrane leaflet. These changes were accompanied by modified membrane bulk fluidity. All three statins reduced the expression of the raft marker protein flotillin. Alterations in transbilayer cholesterol distribution have been suggested as the underlying mechanism that forces amyloidogenic processing of APP in AD. Thus, our data give some first insight in the mode of action of statins to reduce the prevalence of AD in clinical trials. PMID- 12628481 TI - The effect of sesquiterpene lactones on the release of human neutrophil elastase. AB - Sesquiterpene lactones (SLs) are natural products responsible for the anti inflammatory activity of a variety of medicinal plants, mainly from the Asteraceae family. Here, we investigated whether they also influence the process of exocytosis of pro-inflammatory enzymes, such as the human neutrophil elastase (HNE). Altogether, eight structurally different SLs from the eudesmanolide, guaianolide, pseudoguaianolide, and germacranolide type were studied. Neutrophils were isolated from fresh human blood. After pre-incubation with different concentrations of the respective SL and cytochalasin B, the exocytosis of elastase was initiated either by platelet activating factor or N-formyl-methionyl leucyl-phenylalanine. Inhibition of HNE release was measured by p-nitroaniline formation. The SLs exhibited an inhibitory effect on elastase release from neutrophils challenged either by platelet activating factor or N-formyl-methionyl leucyl-phenylalanine. Concentration-response curves were recorded and the IC(50) values ranged from 2 to 30 microM. Studies on isolated HNE showed that a selective direct inhibition on HNE can be excluded. Interestingly, the inhibitory activity did not correlate with the number of alpha,beta-unsaturated carbonyl functions. The structure-activity relationship and the molecular mechanism are discussed. PMID- 12628480 TI - Inhibition of the NF-kappaB signaling pathway mediates the anti-inflammatory effects of petrosaspongiolide M. AB - Petrosaspongiolide M (PT) is a potent secretory phospholipase A(2) inhibitor and anti-inflammatory agent. This marine metabolite reduced the production of nitrite, prostaglandin E(2), and tumor necrosis factor-alpha in the mouse air pouch injected with zymosan. These effects were also observed in mouse peritoneal macrophages stimulated with zymosan. Inhibition of these inflammatory mediators was related to reductions in inducible nitric oxide synthase, cyclo-oxygenase-2, and tumor necrosis factor-alpha expression. Since nuclear factor-kappaB (NF kappaB) appears to play a central role in the transcriptional regulation of these proteins by macrophages, we investigated the effects of PT on this transcription factor. We found that PT was a potent inhibitor of the NF-kappaB pathway since at 1 microM it strongly decreased NF-kappaB-DNA binding in response to zymosan, in mouse peritoneal macrophages. Our study also indicated that PT could interfere with a key step in NF-kappaB activation, the phosphorylation of IkappaBalpha, resulting in inhibition of IkappaBalpha degradation. The control of a wide range of mediators by PT suggests a potentially wide therapeutic spectrum for this marine metabolite in inflammatory conditions. PMID- 12628482 TI - Heme oxygenase-1 induction and regulation in unstimulated mouse peritoneal macrophages. AB - Heme oxygenase-1 (HO-1) is a stress protein induced by a variety of stimuli in inflammatory cells. This study was set up to investigate the induction of this protein in unstimulated macrophages. Resident mouse peritoneal macrophages purified by adhesion and cultured in basal conditions strongly induced HO-1 in a time-dependent manner, with a peak at 20 hr. At the same time, low levels of nitrite accumulated in the culture medium and expression of nitric oxide synthase 2 (NOS-2) and NOS-3 protein was detected. Inhibition of NO production and/or NOS expression by incubating macrophages with different drugs inhibiting NOS activity or modulating the redox state of the cell, such as N-acetylcysteine (NAC) resulted in inhibition of HO-1 expression, suggesting that NO is an endogenous mediator of this stress response. In conclusion, mouse peritoneal macrophages cultured in basal conditions develop an adaptive response with up-regulation of HO-1 as a very sensitive marker of oxidative stress. PMID- 12628483 TI - "Multivalent" saccharides: development of new approaches for inhibiting the effects of glycosphingolipid-binding pathogens. AB - A number of diseases are initiated by the adherence of viruses, bacteria, or bacterial toxins to cell surface carbohydrates, a number of which are components of glycosphingolipids (GSLs). Studies of the binding of lectins indicated that many adhered weakly to monomeric carbohydrate ligands. The seminal observation that lectins adhered more strongly to a ligand with multiple carbohydrate binding sites initiated a plethora of studies designed to identify effective "multivalent" carbohydrate ligands for pathogens expressing multiple carbohydrate binding sites. In addition to more completely defining ligand specificity of the carbohydrate-binding pathogen, identification of "multivalent" carbohydrate ligands has led to studies of their efficacy as pathogen inhibitors. This commentary focuses on pathogens that recognize the carbohydrate portion of GSLs. Because many GSL-binding pathogens have been shown to bind "multivalent" saccharides, approaches for identifying and preparing them as well as methods for characterizing their effectiveness as ligands are reviewed. Also discussed are areas of promise that should be investigated and pitfalls that might be encountered in the development of "multivalent" saccharides as pharmacologic agents. PMID- 12628484 TI - Daidzein enhances osteoblast growth that may be mediated by increased bone morphogenetic protein (BMP) production. AB - Daidzein, a natural isoflavonoid found in Leguminosae, has received increasing attention because of its possible role in the prevention of osteoporosis. In the present investigation, primary osteoblastic cells isolated from newborn Wistar rats were used to investigate the effect of this isoflavonoid on osteoblasts. Daidzein (2-50 microM) increased the viability (P<0.05) of osteoblasts by about 1.4-fold. In addition, daidzein (2-100 microM) increased the alkaline phosphatase activity and osteocalcin synthesis (P<0.05) of osteoblasts by about 1.4- and 2.0 fold, respectively. Alkaline phosphatase and osteocalcin are phenotypic markers for early-stage differentiated osteoblasts and terminally differentiated osteoblasts, respectively. Our results indicated that daidzein stimulated osteoblast differentiation at various stages (from osteoprogenitors to terminally differentiated osteoblasts). We also investigated the effect of daidzein on bone morphogenetic protein (BMP) production in osteoblasts that display the mature osteoblast phenotype. The results indicated that BMP2 synthesis was elevated significantly in response to daidzein (the mRNA increased 5.0-fold, and the protein increased 7.0-fold), suggesting that some of the effects of daidzein on the cell may be mediated by the increased production of BMPs by the osteoblasts. In conclusion, daidzein has a direct stimulatory effect on bone formation in cultured osteoblastic cells in vitro, which may be mediated by increased production of BMPs in osteoblasts. PMID- 12628485 TI - Transcriptional suppression of the adrenal cortical peripheral-type benzodiazepine receptor gene and inhibition of steroid synthesis by ginkgolide B. AB - Treatment of rats and adrenocortical cells with ginkgolide B (GKB), a purified component of Ginkgo biloba leaf extracts, reduces the mRNA, protein, and ligand binding levels of the adrenal peripheral-type benzodiazepine receptor (PBR), a mitochondrial cholesterol-binding protein, leading to decreased corticosteroid synthesis. In the Y1 adrenocortical cell line, GKB reduced both PBR levels and cyclic AMP-induced steroid formation. In these cells, GKB, but not various steroids and vitamins, reduced the expression of a reporter gene driven by the DNA sequence -624/-513 relative to the transcription start site of the PBR encoding gene. GKB treatment did not affect the SV40 promoter and increased the cytochrome P450 17alpha-hydroxylase gene promoter driven expression of the reporter gene. Electrophoretic mobility shift assays (EMSAs) indicated the presence of a functional transcriptional element bound to the -624/-513 DNA fragment. This GKB-induced inhibition of PBR was mediated by an interaction with a transcription factor that binds to the -636/-616 PBR-promoter region. Deletion or mutation of this sequence eliminated the DNA-protein interaction and the inhibitory effect of GKB on PBR gene transcription. This DNA-binding protein could be detected in nuclear extracts of rat brain, liver, and testis, but not kidney. It is also present in the human adrenal glands. However, the inhibitory effect following GKB treatment could be seen only in the adrenal glands. These results demonstrate that the GKB-activated inhibition of glucocorticoid production is due to a specific transcriptional suppression of the adrenal PBR gene and suggest that GKB might serve as a pharmacological tool to control excess glucocorticoid formation. PMID- 12628486 TI - Melatonin differentially modulates the expression and function of the hMT1 and hMT2 melatonin receptors upon prolonged withdrawal. AB - Melatonin is synthesized and released following a circadian rhythm and reaches its highest blood levels during the night. It relays signals of darkness to target tissues involved in regulating circadian and seasonal rhythms. Here, we report the expression of human melatonin receptors type 1 and 2 (hMT(1) and hMT(2), respectively) in Chinese hamster ovary (CHO) cells following exposure to melatonin treatments mimicking the amplitude (400 pM) and duration (8 hr) of the nightly melatonin peak and upon withdrawal. Exposure of CHO-MT(1) cells to melatonin (400 pM) for 0.5, 1, 2, 4, and 8 hr significantly increased specific 2 [125I]iodomelatonin (500 pM) binding to hMT(1) melatonin receptors upon 16-hr withdrawal. However, the same treatment did not affect the expression of hMT(2) melatonin receptors. The increase in specific 2-[125I]iodomelatonin (500 pM) binding (162+/-29%, N=3, P<0.05) 16 hr after melatonin withdrawal was parallel to increases in hMT(1) melatonin receptor mRNA (231+/-33%, N=4, P<0.05). This effect was due to an increase in the total number of hMT(1) receptors [B(max) 833+/-97 fmol/mg protein (N=3), control; 1449+/-41 fmol/mg protein (N=3), treated], with no change in binding affinity. The melatonin-mediated increase in MT(1) melatonin receptor expression upon withdrawal was not mediated through either a direct effect of the hormone in the promoter's vector or in the rate of mRNA degradation. In conclusion, melatonin differentially regulates the expression of its own receptors, which may have important implications in the transduction of dark signals in vivo. PMID- 12628487 TI - Protein glutathiolation in human blood. AB - Glutathione (GSH) exists in both free and protein-bound (glutathiolated) forms (GSSP). Protein glutathiolation may represent an important post-translational regulatory mechanism for proteins. However, there are little data regarding the regulation of glutathiolation in blood. Our objectives were to examine GSSP levels of human blood by determining the distribution and variability of blood GSSP, as well as its relationship to free GSH and hemoglobin in healthy adults. To this end, we used a newly modified method allowing for rapid analysis of both GSH and GSSP in blood. GSSP was found in red cells with levels ranging from 4 to 27% of total (free+bound) GSH (mean+/-SD: 12.1+/-4.5%) with a concentration of 0.13+/-0.05 microEq GSH/mL (mean+/-SD). No correlations were observed between GSSP and either GSH (r=-0.085) or hemoglobin (r=0.086). Together these results suggest that the extent of protein glutathiolation in blood is substantial ( approximately 0.1 mmol/L). While the interindividual variation in GSSP is large (34%), its levels are apparently not regulated by GSH content. PMID- 12628488 TI - Minimally modified phosphodiester antisense oligodeoxyribonucleotide directed against the multidrug resistance gene mdr1. AB - In the perspective of reversing multidrug resistance through antisense strategy while avoiding non-antisense effects of all-phosphorothioate oligonucleotides which non-specifically bind to proteins, a minimally modified antisense phosphodiester oligodeoxyribonucleotide has been designed against mdr1, one of the multidrug resistance genes. Its stability in lysates prepared from NIH/3T3 cells transfected with the human mdr1 gene has already been demonstrated. Confocal microspectrofluorometry using a fluorescence resonance energy transfer technique allowed its stability inside living cells to be proven. Its internalization into the cells was achieved with different delivery agents (addition of a cholesteryl group, Superfect or an amphotericin B cationic derivative) and has been followed by fluorescence imaging. For each of the delivery systems, Western blotting allowed its antisense efficiency to be compared to that of an all-phosphorothioate antisense oligonucleotide. No antisense efficiency was demonstrated for the minimally modified ODN when internalized with Superfect. In both other cases, the best extinction of the P glycoprotein expression has always been achieved with the all-phosphorothioate antisense. While the difference was significant in the case the amphotericin B derivative was used as delivery agent (20% remaining protein expression with the all-phosphorothioate vs. 40% with the minimally modified antisense), it was negligible for the cholesterol conjugates (2% vs. 6%). It is of great interest to prove that an almost all-phosphodiester oligonucleotide can be an efficient antisense against an overexpressed gene. The reduction of non-antisense effects as non-specific binding to proteins are of importance in the case relatively high ODN concentrations are used, which can prove to be necessary in the case of overexpressed genes. PMID- 12628489 TI - Apoptotic cell death induction by F 11782 a novel dual catalytic inhibitor of topoisomerases I and II. AB - F 11782 (2",3"-bis-pentafluorophenoxyacetyl-4",6"ethylidene-beta-D-glucoside of 4'-phosphate-4'-dimethylepipodophyllotoxin-2N-methyl glucamine salt), is a novel dual catalytic inhibitor of topoisomerases I and II characterised by marked in vivo antitumour activity, which also proved cytotoxic and exhibited DNA damaging properties in vitro. Mechanisms associated with this cell killing by F 11782 have been examined in P388 leukaemia cells. Treatment with F 11782 resulted in a dose dependent DNA fragmentation coupled with the characteristic morphological features of apoptosis. Apoptosis-inducing concentrations of F 11782 induced caspases-3/7 activation accompanied by proteolytic cleavage of poly(ADP-ribose) polymerase, which could be inhibited by the caspase inhibitor acetyl-Asp-Glu-Val Asp-aldehyde. In addition, F 11782-induced apoptosis in P388 cells was associated with an increased expression of the pro-apototic Bax protein, without significant changes in the level of the anti-apoptotic Bcl-2 protein, and with modification at the mitochondrial membrane function. These results indicate that F 11782 leads to apoptosis through a caspase-3/7 dependent mechanism and suggest that the so called "mitochondrial pathway" is implicated in F 11782-induced apoptosis in P388 cells. PMID- 12628490 TI - The role of multidrug resistance proteins MRP1, MRP2 and MRP3 in cellular folate homeostasis. AB - Previously, we reported that the multidrug resistance proteins MRP1, MRP2 and MRP3 confer resistance to therapeutic antifolates by mediating their cellular extrusion. We now determined whether MRPs also play a role in controlling cellular homeostasis of natural folates. In MRP1, MRP2 and MRP3-transfected 2008 human ovarian carcinoma cells total cellular folate content was 32-38% lower than in 2008 cells (105+/-14pmolfolate/mgprotein) when grown in medium containing 2.3 microM folic acid (FA). Under these conditions cellular growth rates were not compromised. However, when cells were challenged under folate-depleted conditions with a short exposure (4 hr) to FA or leucovorin, MRP1 and MRP3 overexpressing cells were impaired in their growth. In contrast to wild-type cells, MRP1 transfected cells retained only 60% of the maximum growth when exposed to 500 nM leucovorin or 500 microM FA. For 2008/MRP1 and 2008/MRP3 cells FA growth stimulation capacity was dramatically decreased when, during a 4 hr exposure, metabolism into rapidly polyglutamatable and retainable dihydrofolate was blocked by the dihydrofolate reductase inhibitor trimetrexate. To retain growth under such conditions MRP1 overexpressing cells required much higher concentrations of FA (EC(50) > 500 microM) compared to 2008 cells (EC(50): 12 microM). These results suggest that down- and up-regulation of MRP1 (and MRP3) expression can influence cellular folate homeostasis, in particular when cellular retention by polyglutamylation of folates is attenuated. PMID- 12628491 TI - Inhibition of 15-lipoxygenases by flavonoids: structure-activity relations and mode of action. AB - We have recently reported that flavonoids of cocoa inhibit the mammalian 15 lipoxygenase-1-a catalyst of enzymatic lipid peroxidation. To elucidate the structure-activity relationship of the inhibitory effect, we investigated the effects of 18 selected flavonoids of variable structure on pure rabbit reticulocyte and soybean 15-lipoxygenases using linoleic acid as substrate. Moreover, the inhibition by quercetin was studied in detail to gain insight into the mode of action. Quercetin was found to modulate the time-course of the reaction of both lipoxygenases by three distinct effects: (i) prolongation of the lag period, (ii) rapid decrease in the initial rate after the lag phase was overcome, (iii) time-dependent inactivation of the enzyme during reaction but not in the absence of substrate. A comparison of the IC(50) for the rapid inhibition of rabbit reticulocyte 15-lipoxygenase-1 revealed that (i) the presence of a hydroxyl group in the flavonoid molecule is not essential, (ii) a catechol arrangement reinforces the inhibitory effect, (iii) in the presence of a catechol arrangement the inhibitory potency inversely correlates with the number of hydroxyl groups, (iv) a 2,3-double bond in the C ring strengthens the inhibitory effect. The flavone luteolin turned out to be the most potent inhibitor of the mammalian enzyme with an IC(50) of 0.6 microM followed by baicalein (1 microM) and fisetin (1.5 microM). PMID- 12628492 TI - ETA receptor-mediated Ca2+ mobilisation in H9c2 cardiac cells. AB - Expression and pharmacological properties of endothelin receptors (ETRs) were investigated in H9c2 cardiomyoblasts. The mechanism of receptor-mediated modulation of intracellular Ca(2+) concentration ([Ca(2+)](i)) was examined by measuring fluorescence increase of Fluo-3-loaded cells with flow cytometry. Binding assays showed that [125I]endothelin-1 (ET-1) bound to a single class of high affinity binding sites in cardiomyoblast membranes. Endothelin-3 (ET-3) displaced bound [125I]ET-1 in a biphasic manner, in contrast to an ET(B) selective agonist, IRL-1620, that was ineffective. The ET(B)-selective antagonist, BQ-788, inhibited [125I]ET-1 binding in a monophasic manner and with low potency. An ET(A)-selective antagonist, BQ-123, competed [125I]ET-1 binding in a monophasic manner. This antagonist was found to be 13-fold more potent than BQ-788. Immunoblotting analysis using anti-ET(A) and -ET(B) antibodies confirmed a predominant expression of the ET(A) receptor. ET-1 induced a concentration dependent increase of Fluo-3 fluorescence in cardiomyoblasts resuspended in buffer containing 1mM CaCl(2). Treatment of cells with antagonists, PD-145065 and BQ-123, or a phospholipase C-beta inhibitor, U-73122, abolished ET-1-mediated increases in fluorescence. The close structural analogue of U-73122, U-73343, caused a minimal effect on the concentration-response curve of ET-1. ET-3 produced no major increase of Fluo-3 fluorescence. Removal of extracellular Ca(2+) resulted in a shift to the right of the ET-1 concentration-response curve. Both the L-type voltage-operated Ca(2+) channel blocker, nifedipine, and the ryanodine receptor inhibitor, dantrolene, reduced the efficacy of ET-1. Two protein kinase C inhibitors reduced both potency and efficacy of ET-1. Our results demonstrate that ET(A) receptors are expressed and functionally coupled to rise of [Ca(2+)](i) in H9c2 cardiomyoblasts. ET-1-induced [Ca(2+)](i) increase is triggered by Ca(2+) release from intracellular inositol 1,4,5-trisphosphate gated stores; plasma membrane Ca(2+) channels and ryanodine receptors participate in sustaining the Ca(2+) response. Regulation of channel opening by protein kinase C is also involved in the process of [Ca(2+)](i) increase. PMID- 12628493 TI - Characterization of the molecular interactions of interleukin-8 (CXCL8), growth related oncogen alpha (CXCL1) and a non-peptide antagonist (SB 225002) with the human CXCR2. AB - Neutrophil recruitment to inflammatory sites is mediated by two related receptors: CXC chemokine receptors 1 (CXCR1) and 2 (CXCR2). Both receptors share two ligands, interleukin-8 (CXCL8) and GCP-2 (CXCL6), whereas several chemokines, including growth related oncogen alpha (CXCL1) and a non-peptide antagonist (SB 225002) are specific for CXCR2. The objective of this study was to map the different amino acids involved in the binding and activation/inhibition of human CXCR2. This was performed by exchanging non-conserved amino acids of CXCR2 with their counterparts in CXCR1. The mutants generated showed that: (a) for CXCL8 binding, the N-terminus of CXCR1 and the second extra-cellular loop of CXCR2 are determinant, the N-terminus of CXCR2 is not sufficient and the transmembrane domain seven is probably involved; (b) for CXCL1, the N-terminus of CXCR2 is necessary but not sufficient for binding. The activation study indicated that amino acids critical for activation are not necessarily involved in binding process. Finally, the mechanism of binding of a non-peptide antagonist on CXCR2 was investigated: it occurred through epitopes (a) which were disseminated within the receptor, (b) which differed according to the use of CXCL8 or CXCL1 as a competitor and (c) which did not necessarily overlap with agonist binding sites. We also showed that inhibition of binding and inhibition of activation involved different amino acids. PMID- 12628494 TI - An oxidative mechanism of interferon induced priming of the Fas pathway in Fanconi anemia cells. AB - Hematopoietic progenitor cells from children with Fanconi anemia of the C complementation group (FA-C) are excessively apoptotic and hypersensitive to various extracellular cues including Fas-ligand, tumor necrosis factor-alpha and double-stranded RNA. Interferon (IFN)-gamma is known to augment apoptotic responses of these factors. The "priming" effect of IFN-gamma is not fully explained. In view of the strong evidence that FA cells are intolerant of oxidative stress, we tested the notion that IFN-priming involves the induction of reactive oxygen species (ROS) in two FA-C B-lymphocyte cell lines and in peripheral blood neutrophils and mononuclear cells of FA patients. We also investigated whether the combination of IFN-gamma and Fas created an intracellular environment that promoted apoptosis. Significantly lower doses of IFN-gamma induced ROS accumulation in neutrophils and mononuclear cell of FA patients compared to cells of normal individuals. Enhanced ROS accumulation and decreased intracellular glutathione levels were observed in FA-C B-cell lines primed with IFN-gamma and treated with agonistic anti-Fas antibody than in isogenic control cells corrected with FANCC. The above treatment also induced caspase-3 and -8 activation as well as apoptosis. That antioxidants reduced the priming effect of IFN-gamma in Fas and IFN-gamma-treated FA lymphoblast cells, demonstrates that ROS represent a critical effector mechanism for the exaggerated responses to IFN-gamma characteristic of FA-C cells. PMID- 12628495 TI - Toxicogenomics of bromobenzene hepatotoxicity: a combined transcriptomics and proteomics approach. AB - Toxicogenomics is a novel approach integrating the expression analysis of thousands of genes (transcriptomics) or proteins (proteomics) with classical methods in toxicology. Effects at the molecular level are related to pathophysiological changes of the organisms, enabling detailed comparison of mechanisms and early detection and prediction of toxicity. This report addresses the value of the combined use of transcriptomics and proteomics technologies in toxicology. Acute hepatotoxicity was induced in rats by bromobenzene administration resulting in depleted glutathione levels and reduced average body weights, 24hr after dosage. These physiological symptoms coincided with many changes of hepatic mRNA and protein content. Gene induction confirmed involvement of glutathione-S-transferase isozymes and epoxide hydrolase in bromobenzene metabolism and identified many genes possibly relevant in bromobenzene toxicity. Observed glutathione depletion coincided with induction of the key enzyme in glutathione biosynthesis, gamma-glutamylcysteine synthetase. Oxidative stress was apparent from strong upregulation of heme oxygenase, peroxiredoxin 1 and other genes. Bromobenzene-induced protein degradation was suggested from two dimensional gel electrophoresis, upregulated mRNA levels for proteasome subunits and lysosomal cathepsin L, whereas also genes were upregulated with a role in protein synthesis. Both protein and gene expression profiles from treated rats were clearly distinct from controls as shown by principal component analysis, and several proteins found to significantly change upon bromobenzene treatment were identified by mass spectrometry. A modest overlap in results from proteomics and transcriptomics was found. This work indicates that transcriptomics and proteomics technologies are complementary to each other and provide new possibilities in molecular toxicology. PMID- 12628496 TI - Flavonoids inhibit myelin phagocytosis by macrophages; a structure-activity relationship study. AB - Demyelination is a characteristic hallmark of the neuro-inflammatory disease multiple sclerosis. During demyelination, macrophages phagocytose myelin and secrete inflammatory mediators that worsen the disease. Here, we investigated whether flavonoids, naturally occurring immunomodulating compounds, are able to influence myelin phagocytosis by macrophages in vitro. The flavonoids luteolin, quercetin and fisetin most significantly decreased the amount of myelin phagocytosed by a macrophage cell line without affecting its viability. IC(50) values for these compounds ranged from 20 to 80 microM. The flavonoid structure appeared to be essential for observed effects as flavonoids containing hydroxyl groups at the B-3 and B-4 positions in combination with a C-2,3 double bond were most effective. The capacity of the various flavonoids to inhibit phagocytosis correlated well with their potency as antioxidant, which is in line with the requirement of reactive oxygen species for the phagocytosis of myelin by macrophages. Our results implicate that flavonoids may be able to limit the demyelination process during multiple sclerosis. PMID- 12628497 TI - Calpain inhibitor (BSF 409425) diminishes ischemia/reperfusion-induced damage of rabbit heart mitochondria. AB - Calpains are involved in ischemia/reperfusion-induced changes of myocard. To obtain information on the action of calpain on mitochondria, the effect of a new developed calpain inhibitor (CI) BSF 409425 on the ischemia/reperfusion-induced damage of rabbit heart mitochondria was investigated. Rabbit hearts were subjected to 45 min of global ischemia followed by 60 min of reperfusion in the presence or absence of 10nM CI. Mitochondrial properties were characterized by skinned fiber technique with pyruvate+malate as substrates. In the presence of CI, the decrease of state 3 respiration and the increase of state 4 respiration after ischemia and reperfusion were clearly smaller than without CI resulting in significantly smaller changes of respiratory control index, too. Ischemia/reperfusion-caused leaks in mitochondrial inner and outer membranes were diminished by CI. It is concluded that mitochondria are a target of calpain which reinforces the damage of oxidative phosphorylation and mitochondrial membranes during ischemia/reperfusion. PMID- 12628498 TI - Dietary and medicinal antimutagens and anticarcinogens: molecular mechanisms and chemopreventive potential--highlights of a symposium. AB - Antimutagens and anticarcinogens are common amongst many traditional herbal remedies and dietary therapies. With increased understanding of the mechanistic basis of cancer development and cancer prevention, we are now better aware of ways in which some of these traditional remedies may act at the cellular or subcellular levels. This special issue features some of the highlights of the conference on this topic that was held in Seoul on October 17-19, 2001. PMID- 12628499 TI - Methodological considerations for characterizing potential antioxidant actions of bioactive components in plant foods. AB - The study of free radicals and antioxidants in biology is producing medical revolution that promises a new age of health and disease management. From prevention of the oxidative reactions in foods, pharmaceuticals and cosmetics to the role of reactive oxygen species (ROS) in chronic degenerative diseases including cancer, autoimmune, inflammatory, cardiovascular and neurodegenerative (e.g. Alzheimer's disease, Parkinson's disease, multiple sclerosis, Downs syndrome) and aging challenges continue to emerge from difficulties associated with methods used in evaluating antioxidant actions in vivo. Our interest presently is focused on development of neurodegeneration models based on the integrity of neuronal cells in the central nervous system and how they are protected by antioxidants when challenged by neurotoxins as well as Fenton chemistry models based on the profile of polyunsaturated fatty acids (PUFAs) for the assessment of antioxidant actions in vivo. Use continues to be made of several in vitro analytical tools to characterise the antioxidant propensity of bioactive compounds in plant foods and supplements. For example, the oxygen radical absorbance capacity (ORAC), ferric reducing antioxidant power (FRAP), total oxidant scavenging capacity (TOSC), the deoxyribose assay, assays involving oxidative DNA damage, assays involving reactive nitrogen intermediates (e.g. ONOO(-)), Trolox equivalent antioxidant capacity (TEAC) and the 2,2-diphenyl-1 picrylhydrazyl (DPPH) assay. There is need to agree governance on in vitro antioxidant methods based on an understanding of the mechanisms involved. Because some of the assays are done in non-physiological pH values, it is impossible to extrapolate the results to physiological environment. The consensus of opinion is that a mix of these tools should be used in assessing the antioxidant activities in vitro. The proof of bio-efficacy must emanate from application of reliable in vivo models where markers of baseline oxidative damage are examined from the standpoint of how they are affected by changes in diet or by antioxidant supplements. PMID- 12628500 TI - Antimutagenicity of sodium selenite in Chinese hamster V79 cells exposed to azoxymethane, methylmethansulphonate and hydrogen peroxide. AB - In the last 10 years, there is an increasing interest in selenium (Se) because of its environmental, biological, and toxicological importance, and in particular, because of its antioxidant properties. However, inspite of extensive studies, the optimal concentration of Se to be used for its beneficial effects in not yet known. In addition, the mechanisms of Se antioxidant property require further study. We report on the effects of various mutagens/carcinogens such as azoxymethane, methylmethanesulphonate, and hydrogen peroxide on Chinese V79 hamster cells, in presence of sodium selenite in the concentration of 0.5 microM. We found that Se reduced the genotoxic effect of these mutagens/carcinogens. We also investigated enzymatic activities of glutathione peroxidase, catalase, superoxide dismutase, and glutathione S-transferase, in order to understand the Se involvement in the detoxification of free radicals. Sodium selenite increased the activities of glutathione peroxidase and catalase. PMID- 12628502 TI - Antimutagenic effects of doenjang (Korean fermented soypaste) and its active compounds. AB - Doenjang (Korean fermented soypaste) is one of the important fermented foods of Korea. Doenjang has been traditionally manufactured from meju, which is fermented rectangular shape molded from crushed cooked soybeans. The main microorganisms involved for meju fermentation are Bacillus subtilis and molds such as Rizopus sp., Mucor sp., and Aspergillus sp. We have already reported that doenjang is safe from mycotoxin, especially, aflatoxin contamination due to the manufacturing process of the doenjang. We have demonstrated that the doenjang extracts showed strong antimutagenic activities against various carcinogens/mutagens including aflatoxin B(1). The traditionally fermented soypaste, doenjang showed higher antimutagenic activity than the raw soybeans, cooked soybeans, meju and other fermented soybeans in the Ames test. The active compounds that were identified are genistein, linoleic acid, beta-sitosterol glucoside, soyasaponin, etc. The active compounds exhibited strong antimutagenic activities in the Ames test, SOS chromotest and Drosophila wing spot test. More genistein was formed during the doenjang fermentation from genistin in the soybeans. Genistein and linoleic acid were the most effective active compounds found in doenjang. PMID- 12628501 TI - Anticlastogenic, antigenotoxic and apoptotic activity of epigallocatechin gallate: a green tea polyphenol. AB - Modulation of events characteristic of carcinogenesis or of cancer cells is being emphasized as a rational strategy to control cancer. Green tea polyphenol epigallocatechin gallate (EGCG) has been shown to be highly active as a cancer chemopreventive agent. Certain cellular and molecular events relevant to carcinogenesis are also modified by EGCG. The present investigation was carried out to examine the effects of EGCG on the cytogenetic change and DNA damage induced by toxicant H(2)O(2) and carcinogen N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in Chinese hamster V-79 cells in culture. Cytogenetic change as evident by the formation of micronuclei and DNA damage in the form of comet tail length during single cell gel electrophoresis was found to be significantly suppressed by EGCG in a dose dependent manner. Cells preincubated with EGCG were protected from subsequent damage by the genotoxic agents. Apoptosis, a highly organized physiological mechanism to eliminate injured or abnormal cells, is also implicated in multistage carcinogenesis. Initiated cells, cells at promotional stage or fully transformed cells can be eliminated through apoptosis. It was observed that EGCG suppressed growth and proliferation of K-562 cells derived from human chronic myelogenic leukemia. Morphological features of treated cells and characteristic DNA fragmentation revealed that the cytotoxicity was due to induction of apoptosis. This was mediated by activation of caspase 3 and caspase 8. Results show that EGCG not only protects normal cells against genotoxic hazard but also eliminate cancer cells through induction of apoptosis. PMID- 12628503 TI - Inhibition of human breast cancer growth by GCP (genistein combined polysaccharide) in xenogeneic athymic mice: involvement of genistein biotransformation by beta-glucuronidase from tumor tissues. AB - The role of beta-glucuronidase in genistein biotransformation was investigated in a human breast cancer MDA-MB-231 xenogeneic athymic mouse model. Genistein combined polysaccharide (GCP), a genistein aglycone rich functional food supplement was used in these experiments. Tumor-bearing mice were subjected to oral administration of GCP for 28 days. GCP treatment significantly inhibited tumor growth. Induction of apoptosis by GCP treatment was related to activation of cleavage of poly(ADP-ribose)polymerase, induction of the p21 protein expression and reduction of cyclin B1 expression in the tumor tissues. Genistein exists as a glucuronide conjugate in normal organ tissues, and the conjugated genistein lacks the physiological activity of the aglycone. Tumor tissues contain large amounts of beta-glucuronidase, the enzyme that converts the genistein beta glucuronide conjugate into genistein aglycone. The resulting genistein aglycone exerts its chemopreventive activities, including the induction of apoptosis in tumor tissues, and, finally, leads to tumor growth inhibition. PMID- 12628504 TI - Experimental and epidemiological evidence on non-organ specific cancer preventive effect of Korean ginseng and identification of active compounds. AB - Panax ginseng C.A. Meyer has been the most highly recognized medicinal herb in the Orient. The prolonged administration of red ginseng extract significantly inhibits the incidence of hepatoma and also proliferation of pulmonary tumors induced by aflatoxin B(1) and urethane. Statistically significant anticarcinogenic effects were in aged or heat treated extracts of ginseng and red ginseng made by steaming in a 9 weeks medium-term anticarcinogenicity test using benzo[a]pyrene. In case-control studies, odds ratios (OR) of the cancer of lip, oral cavity and pharynx, larynx, lung, esophagus, stomach, liver, pancreas, ovary, and colorectum were significantly reduced. As to the type of ginseng, the ORs for cancer were reduced in user of fresh ginseng extract intakers, white ginseng extract, white ginseng powder, and red ginseng. In a cohort study with 5 years follow-up conducted in a ginseng cultivation area, ginseng users had a decreased relative risk (RR) compared with non-users. The relative risks (RRs) of ginseng users were decreased in gastric cancer and lung cancer. These findings strongly suggest that Panax ginseng C.A. Meyer cultivated in Korea has non-organ specific cancer preventive effects against various cancers. To investigate the active components for cancer prevention, several fractions of fresh and red ginseng and four semi-synthetic ginsenoside Rh(1), Rh(2), Rg(3) and Rg(5), the major saponin components in red ginseng, were prepared among the ginsenosides. By using Yun's model, Rg(3) and Rg(5) showed statistically significant reduction of lung tumor incidence and Rh(2) had a tendency to decrease the incidence. In conclusion, these results strongly suggested that Panax ginseng C.A. Meyer cultivated in Korea is a non-organ specific cancer preventive against human cancers and also indicated that the anticarcinogenicity or human cancer preventive effect of Panax ginseng is due to ginsenoside Rg(3), Rg(5) and Rh(2). PMID- 12628506 TI - Molecular mechanisms of cardioprotection by a novel grape seed proanthocyanidin extract. AB - Free radicals and oxidative stress play a crucial role in the pathophysiology of a broad spectrum of cardiovascular diseases including congestive heart failure, valvular heart disease, cardiomyopathy, hypertrophy, atherosclerosis and ischemic heart disease. We have demonstrated that IH636 grape seed proanthocyanidin extract (GSPE) provides superior antioxidant efficacy as compared to Vitamins C, E and beta-carotene. A series of studies were conducted using GSPE to demonstrate its cardioprotective ability in animals and humans. GSPE supplementation improved cardiac functional assessment including post-ischemic left ventricular function, reduced myocardial infarct size, reduced ventricular fibrillation (VF) and tachycardia, decreased the amount of reactive oxygen species (ROS) as detected by ESR spectroscopy and reduced malondialdehyde (MDA) formation in the heart perfusate. Cardiomyocyte apoptosis detected by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) staining. In concert, the proapoptotic signals mediated by JNK-l and c-fos proteins were also reduced suggesting that the novel cardioprotective properties of GSPE may be at least partially attributed to its ability to block anti-death signaling mediated through the proapoptotic transcription factors and genes such as JNK-1 and c-JUN. In a separate study, GSPE pretreatment significantly inhibited doxorubicin induced cardiotoxicity as demonstrated by reduced serum creatine kinase (CK) activity, DNA damage and histopathological changes in the cardiac tissue of mice. Concentration-dependent efficacy of GSPE was also assessed in a hamster atherosclerosis model. Approximately 49 and 63% reduction in foam cells, a biomarker of early stage atherosclerosis, were observed following supplementation of 50 and 100 mg GSPE/kg body weight, respectively. A human clinical trial was conducted on hypercholesterolemic subjects. GSPE supplementation significantly reduced oxidized LDL, a biomarker of cardiovascular diseases. Finally, a cDNA microarray study demonstrated significant inhibition of inducible endothelial CD36 expression, a novel cardioregulatory gene, by GSPE. These results demonstrate that GSPE may serve as a potential therapeutic tool in promoting cardiovascular health via a number of novel mechanisms. PMID- 12628505 TI - Inhibitory effects of the ginsenoside Rg3 on phorbol ester-induced cyclooxygenase 2 expression, NF-kappaB activation and tumor promotion. AB - Our previous studies demonstrated the anti-oxidant and anti-tumor promotional properties of the methanol extract of heat-processed Panax ginseng C.A. Meyer [Cancer Lett. 150 (2000) 41]. In the present work, we have evaluated anti inflammatory as well as anti-tumor promoting effects of Rg(3), a major ginsenoside derived from heat-processed ginseng. Pretreatment of dorsal skins of female ICR mice with Rg(3) significantly inhibited 12-O-tetradecanoylphorbol-13 acetate (TPA)-induced ornithine decarboxylase activity and 7,12 dimethylbenz[a]anthracene-initiated papilloma formation. In another experiment, Rg(3) pretreatment abrogated the expression of cyclooxygenase-2 in TPA-stimulated mouse skin. Rg(3) also inhibited the TPA-induced activation of the eukaryotic transcription factor, NF-kappaB in both mouse skin and cultured human pro myelocytic leukemia (HL-60) cells. Moreover, Rg(3) exerted potent inhibitory effects on the activation of another transcription factor, activator protein-1 (AP-1) that is responsible for c-jun and c-fos oncogenic transactivation. Based on these findings, it is likely that the anti-tumor promoting activity of Rg(3) is mediated possibly through down-regulation of NF-kappaB and AP-1 transcription factors. PMID- 12628507 TI - Chemoprevention of colon cancer by Korean food plant components. AB - Inducible cyclooxygenase (COX-2) and inducible nitric oxide synthase (iNOS/NOS-2) play pivotal roles as mediators of inflammation involved in early steps of carcinogenesis in certain organs. Therefore, chemoprevention is theoretically possible through inhibition of COX-2 and/or iNOS. In the present study, we examined the chemopreventive effects of indole-3-carbinol (I3C), a constituent of cruciferous vegetables (the family of Cruciferae) such as cabbages, cauliflowers and broccoli on the multiple intestinal neoplasia (Min) genetic mouse model, and on mouse colon carcinogenesis induced by azoxymethane (AOM). The consumption of cruciferous vegetables such as cabbage, broccoli, and Brussels sprouts has been shown to have cancer chemopreventive effects in humans and experimental animals. I3C has been shown to exert a cancer chemopreventive influence in liver, colon, and mammary tissue when given before or concurrent with exposure to a carcinogen. Powdered AIN-76A diets (Harlan Teklad Research Diet, Madison, USA) containing 100 or 300 ppm I3C (group 1 or 2) or the same pellet diets without supplement (group 3) were fed to 6-week-old male C57BL/6J-Apc(Min)(/+) (Min/+) mice (The Jackson Laboratory, Bar Harbor, ME, USA) for 10 weeks. In addition the same diets were given to wild-type normal C57BL/6J-Apc(Min)(/+) littermates after AOM initiation (groups 4-7: 10 mice in each group) for 32 weeks from week 4. At 16 weeks of age, all Min/+ mice (groups 1-3) were sacrificed for assessment of intestinal polyp development. The incidences of the colonic adenomatous polyps in the groups 1-3 were 60% (12/20), 60% (15/25) and 84% (21/25), respectively. A decreasing tendency in multiplicities of the colonic adenomatous polyps in group 1 (I3C 100 ppm; 0.85 +/- 0.22; 61%) and group 2 (I3C 300 ppm; 1.32 +/- 0.28; 94%) was observed when compared with group 3 (control; 1.40 +/- 0.21; 100%). Total number of aberrant crypt foci (ACF)/colon or aberrant crypts (AC)/colon in wild-type mice of group 4 or 5 were decreased significantly compared with those of the AOM alone group (group 6) (P < 0.01). These results suggest that I3C may be a potential chemopreventive agent for colon cancer. PMID- 12628508 TI - Potential functional foods in the traditional Maori diet. AB - The Maori people were early New Zealand settlers of Polynesian descent. The incidence of non-infectious diseases appears to have been low in these people, perhaps in part due to the presence of protective chemical constituents within their food plant supply. Three of the tropical crops they introduced are still eaten here today: the sweet potato or kumara (Ipomoea batatas), the taro (Colocasia esculenta) and the cabbage tree or ti (Cordyline terminalis). Sporamins A and B, the major storage proteins of kumara tubers, act as proteinase inhibitors, and may have other anti-cancer properties. The tubers also contain the anti-coagulant coumarins, scopoletin, aesculetin, and umbelliferone. The corms of taro contain the anthocyanins, cyanidin 3-glucoside, pelargonidin 3 glucoside and cyanidin 3-rhamnoside, reported to have antioxidant and anti inflammatory properties. Anthocyanins are also major components of a so-called "Maori potato", a variety officially known as Ureniki, which has a purple skin and flesh and was widely eaten in the early 1900s. Anthocyanins are also present in ripe berries of the ramarama (Lophomyrtus bullata) and rohutu (Neomyrtus pedunculata). Both the leaves and seeds of the introduced cabbage tree (Cordyline terminalis) and the native Cordyline spp., C. australis, C. indivisa, and C. pumilo, were eaten. The seeds of C. australis, of some Astelia spp., and of hinau (Elaeocarpus dentatus) are good sources of various essential fatty acids, generally regarded as protective against cardiovascular disease. Shoots and leaves from a wide range of native species were traditionally eaten as greens, especially "sow thistle" or puha (Sonchus spp.), reportedly high in Vitamin C and various phenolics. "New Zealand spinach" (Tetragonia tetragonioides or T. expansa) has anti-ulcerogenic activity that has been traced to two cerebrosides and anti-inflammatory activity that has been traced to novel water-soluble polysaccharides, as well as antioxidant phenylpropanoids including caffeic acid. Leaves of the "hen and chickens" fern (Asplenium bulbiferum) contain antioxidant flavonoids such as kaempferol glucosides. Native seaweeds also have useful nutritive properties. PMID- 12628509 TI - New TNF-alpha releasing inhibitors as cancer preventive agents from traditional herbal medicine and combination cancer prevention study with EGCG and sulindac or tamoxifen. AB - Herbal medicines are now attracting attention as potential sources of cancer preventive agents. Using inhibition of tumor necrosis factor-alpha (TNF-alpha) release assay, we studied Acer nikoense, Megusurino-ki in Japanese. Inhibitory potential was found in the leaf extract, and the main active principles were identified as geraniin and corilagin. The IC(50) values for TNF-alpha release inhibition were 43 microM for geraniin and 76 microM for corilagin, whereas that for (-)-epigallocatechin gallate (EGCG), the green tea polyphenol, as control was 26 microM. Furthermore, treatment with geraniin inhibited okadaic acid tumor promotion in a two-stage carcinogenesis experiment on mouse skin. Geraniin and corilagin are present in another well-known Japanese traditional herb, Geranium thunbergii, Genno-shoko in Japanese. Considering seasonal variations of the agents and sites of cultivation of herbs, this paper reviews the significance of geraniin as a new cancer preventive agent. In addition, based on accumulated results of green tea as a cancer preventive, we review two important results with EGCG: the synergistic effects of EGCG with sulindac or tamoxifen on cancer preventive activity in PC-9 cells, and cancer prevention of intestinal tumor development in multiple intestinal neoplasia (Min) mice by cotreatment using EGCG with sulindac. We report here new findings on additional gene expression resulting from cotreatment with EGCG and sulindac in PC-9 cells compared with gene expression by EGCG alone or sulindac alone. Overall, our results indicate that, with the continuing spread of cancer chemoprevention as a fundamental medical strategy, both clinicians and researchers should take a closer look at herbal medicine. PMID- 12628510 TI - No prevention of liver and kidney tumors in Long-Evans Cinnamon rats by dietary curcumin, but inhibition at other sites and of metastases. AB - Long-Evans Cinnamon (LEC) rats, an inbred mutant strain which accumulates copper due to an aberrant copper-transporting ATPase gene, develop acute hepatitis, chronic liver injury and liver tumors as a result of copper-induced oxidative stress, lipid peroxidation and DNA damage. Curcumin, an antioxidant and anti inflammatory agent, has shown anticancer properties in many rodent models. We investigated the modulating role of curcumin in liver and kidney carcinogenesis in LEC rats. Two groups of 4-week-old LEC rats (n = 60 each) were fed either a standard diet (control) or received 0.5% curcumin in the diet for life. In untreated LEC rats, the rate of acute liver failure, the incidence of liver tumors and of kidney tumors were 32, 100 and 10% respectively, which was not altered by curcumin treatment. However, curcumin reduced tumor incidence at other organ sites (15% versus 0%; P = 0.025) and suppressed formation of metastases (18% versus 0%; P = 0.01). Median survival time was decreased from 88.7 to 78.1 weeks in curcumin-treated rats (P = 0.002). The lack of chemoprevention of liver and kidney tumors in LEC rats by curcumin may be caused by enhanced toxicity and oxidative stress due to excess copper. We conclude that curcumin should be contra indicated for patients suffering from inherited and acquired metal storage diseases that include patients with hepatitis C virus infection. PMID- 12628511 TI - Mechanisms and applications of non-steroidal anti-inflammatory drugs in the chemoprevention of cancer. AB - Biological and chemical irritants can be the cause of irritation in a variety of organ sites. It is becoming well understood that chronic irritation in any form can initiate and accelerate the cancer process in these same organs. This understanding comes in part from the many epidemiologic studies which point out that chronic inflammation correlates with increased risk of developing cancer in that organ which is affected. One of the hallmarks of chronic irritation is the increased activity in the arachidonic acid pathway which provides many of the necessary inflammatory biochemical mediators to this process. Arachidonic acid metabolism diverges down two main pathways, the cyclooxygenase (COX) and the lipoxygenase (LOX) pathways. The COX pathway leads to prostaglandin and thromboxane production and the LOX pathway leads to the leukotrienes (LTs) and hydroxyeicosatetraenoic acids (HETEs). These classes of inflammatory molecules exert profound biological effects which enhance the development and progression of human cancers. A large number of synthetic drugs and natural products have been discovered that block many of these key pathways. Much experimental evidence in animals has shown that inhibition of the key enzymes which drive these pathways can, in fact, prevent, slow or reverse the cancer process. The data are convincing in a number of organ sites including colon, breast, lung, bladder and skin. More recently, double-blinded randomize clinical trials in humans have shown the prevention of colonic polyps by anti-inflammatory agents. These studies have primarily used non-steroidal anti-inflammatory drugs (NSAIDS) which block the COX pathways. Recent preclinical studies indicate that the LOX pathway also may be an important target for cancer prevention strategy. The expression of high levels of these enzymes in cancerous tissues make them an obvious first target for cancer prevention strategies. As newer more specific drugs are developed with few adverse effects this important prevention strategy may become a reality. PMID- 12628512 TI - Molecular mechanism of the chemopreventive effect of resveratrol. AB - Chemoprevention is a promising approach to control human cancer. Resveratrol has been shown to have a potent chemopreventive effect in multiple carcinogenesis models. However, the precise mechanism explaining its anti-carcinogenic effect is not clear. This review summarizes recent studies from our laboratory on the mechanisms of resveratrol's effects. In JB6 cells, resveratrol was found to induce apoptosis and inhibit tumor promoter-induced cell transformation. We also found that resveratrol-induced activation of p53 and resveratrol-induced apoptosis occurred through a p53-dependent pathway. The MAP kinases, ERKs, JNKs, or p38 kinases, are involved in resveratrol-induced activation of p53 and apoptosis. PMID- 12628513 TI - Synergistic suppression of superoxide and nitric oxide generation from inflammatory cells by combined food factors. AB - In contrast to chemopreventive strategies using individual agents, a combination of specified compounds may be effectual to achieve desirable results with higher efficacy and lower toxicity. In the present in vitro study, we examined combinations of agents and assessed which concentrations were appropriate to yield notable synergism. L-N(G)-Monomethyl-L-arginine (L-NMMA), a synthetic inducible nitric oxide synthase (iNOS) inhibitor, and zerumbone, a natural sesquiterpene that suppresses iNOS de novo synthesis, were combined at various concentrations, with the aim to diminish combined lipopolysaccharide- and interferon-gamma-induced nitric oxide generation in a murine macrophage line, RAW264.7. Although the combinatorial effects (CEs) were antagonistic or additive at higher concentrations, significant synergism was obtained at lower concentrations where each agent alone did not cause significant inhibition. Similarly, the CEs were synergistic when (-)-epigallocatechin gallate (EGCG) and genistein were combined at lower concentrations, whereas those of two iNOS inhibitors, L-NMMA and L-N(G)-aminoethyl-L-ornithine, were either additive or antagonistic at all concentrations tested, suggesting that a combination of given agents with different action mechanisms is a prerequisite for synergistic effects. For suppression of phorbol ester-induced superoxide anion radical (O(2)*(-)) generation in differentiated HL-60 cells, the CEs of 1' acetoxycahvicol acetate (ACA), a phenyl propanoid that suppresses O(2)*(-) generation, and O(2)*(-) dismutase were also synergistic, though only at lower concentrations. The CEs of ACA/EGCG were antagonistic or additive, even at low concentrations, suggesting that the signal transduction pathways triggered by these agents are antagonistic. The present findings suggest that individual food phytochemicals have complex interactions that can be antagonistic, additive, and/or synergistic in biological systems, depending upon certain environmental factors including concentrations. Further, these results support and emphasize the concept that combinations of different types of chemicals at low concentrations are one of the essential areas of study for chemopreventive strategies. PMID- 12628514 TI - Mechanism-based in vitro screening of potential cancer chemopreventive agents. AB - Identification and use of effective cancer chemopreventive agents have become an important issue in public health-related research. For identification of potential cancer chemopreventive constituents we have set up a battery of cell- and enzyme-based in vitro marker systems relevant for prevention of carcinogenesis in vivo. These systems include modulation of drug metabolism (inhibition of Cyp1A activity, induction of NAD(P)H:quinone reductase (QR) activity in Hepa1c1c7 murine hepatoma cell culture), determination of radical scavenging (DPPH scavenging) and antioxidant effects (scavenging of superoxide anion-, hydroxyl- and peroxyl-radicals), anti-inflammatory mechanisms (inhibition of lipopolysaccharide (LPS)-mediated nitric oxide (NO) generation by inducible NO synthase (iNOS) in Raw 264.7 murine macrophages, cyclooxygenase-1 (Cox-1) inhibition), and anti-tumor promoting activities (inhibition of phorbol ester induced ornithine decarboxylase (ODC) activity in 308 murine keratinocytes). We have tested a series of known chemopreventive substances belonging to several structural classes as reference compounds for the identification of novel chemopreventive agents or mechanisms. These include organosulfur compounds (phenethylisothiocyanate (PEITC), diallylsulfide, diallyldisulfide), terpenes (limonene, perillyl alcohol, oleanolic acid, 18-beta-glycyrrhetinic acid), short chain fatty acids (sodium butyrate), indoles (indole-3-carbinol), isoflavonoids (quercetin, silymarin, genistein), catechins ((-)-epigallocatechin gallate (EGCG)), simple phenols (ellagic acid, resveratrol, piceatannol, curcumin), pharmaceutical agents (piroxicam, acetylsalicylic acid, tamoxifen), and vitamins/derivatives (ascorbic acid, Trolox). We confirmed known chemopreventive mechanisms of these compounds. Additionally, we could demonstrate the usefulness of our approach by identification of hitherto unknown mechanisms of selected agents. As an example, we detected anti-inflammatory properties of PEITC, based on NF-kappaB-mediated inhibition of NO production. Further, PEITC inhibited phorbol ester-induced superoxide anion radical production in granulocytes, and ODC induction in the 308 cell line. These mechanisms might contribute to the chemopreventive potential of PEITC. PMID- 12628515 TI - Cytochrome P450 1B1: a target for inhibition in anticarcinogenesis strategies. AB - Cytochrome P450 (P450) 1B1 is expressed in a number of human tissues in which cancers occur (e.g. prostate, ovary, uterus, mammary gland). P450 1B1 activates many environmental mutagens and also catalyzes the 4-hydroxylation of estrogens, considered to be an important step in hormonal carcinogenesis. We have examined the activities of several of the major allelic variants of human P450 1B1 in these reactions. Another interest has been the development of chemical inhibitors of P450 1B1. 2-Ethynylpyrene and alpha-naphthoflavone preferentially inhibit human P450 1B1 compared to P450 1A1, which may be present in the same tissue sites. The natural product resveratrol is also an inhibitor of P450 1B1. Further studies with rhapontigenin and synthetic stilbenes led to the discovery of 2,4,3',5'-tetramethoxystilbene, a selective inhibitor of P450 1B1 relative to other P450s. Inhibition is competitive, with a K(i) value of 3nM, and the inhibitor is resistant to metabolism. In addition to blocking 17beta-estradiol 4 hydroxylation, this stilbene also inhibited the activation of heterocyclic amines to mutagens. 2,4,3',5'-Tetramethoxystilbene also suppressed expression of P450 1B1 and growth of human mammary tumor cells. 3,3',4',5,5'-Pentamethoxystilbene was a selective inhibitor of P450 1A1, showing mixed inhibition, and also suppressed P450 1A1 expression in HepG2 cells. Substituted stilbenes may be useful in preventing cancer caused by estrogens and xenobiotics. PMID- 12628516 TI - Development and application of test methods for the detection of dietary constituents which protect against heterocyclic aromatic amines. AB - This article describes the development and use of assay models in vitro (genotoxicity assay with genetically engineered cells and human hepatoma (HepG2) cells) and in vivo (genotoxicity and short-term carcinogenicity assays with rodents) for the identification of dietary constituents which protect against the genotoxic and carcinogenic effects of heterocyclic aromatic amines (HAs). The use of genetically engineered cells expressing enzymes responsible for the bioactivation of HAs enables the detection of dietary factors that inhibit the metabolic activation of HAs. Human derived hepatoma (HepG2) cells are sensitive towards HAs and express several enzymes [glutathione S-transferase (GST), N acetyltransferase (NAT), sulfotransferase (SULT), UDP-glucuronosyltransferase (UDPGT), and cytochrome P450 isozymes] involved in the biotransformation of HAs. Hence these cells may reflect protective effects, which are due to inhibition of activating enzymes and/or induction of detoxifying enzymes. The SCGE assay with rodent cells has the advantage that HA-induced DNA damage can be monitored in a variety of organs which are targets for tumor induction by HAs. ACF and GST-P(+) foci constitute preneoplastic lesions that may develop into tumors. Therefore, agents that prevent the formation of these lesions may be anticarcinogens. The foci yield and the sensitivity of the system could be substantially increased by using a modified diet. The predictive value of the different in vitro and in vivo assays described here for the identification of HA-protective dietary substances relevant for humans is probably better than that of conventional in vitro test methods with enzyme homogenates. Nevertheless, the new test methods are not without shortcomings and these issues are critically discussed in the present article. PMID- 12628517 TI - Direct reduction of N-acetoxy-PhIP by tea polyphenols: a possible mechanism for chemoprevention against PhIP-DNA adduct formation. AB - The chemopreventive effect of tea against 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine (PhIP)-DNA adduct formation and its mechanism were studied. Rats were exposed to freshly prepared aqueous extracts of green tea (3% (w/v)) as the sole source of drinking water for 10 days prior to administration with a single dose of PhIP (10 mg/kg body weight) by oral gavage. PhIP-DNA adducts in the liver, colon, heart, and lung were measured using the 32P-postlabelling technique. Rats pre-treated with tea and given PhIP 20 h before sacrifice had significantly reduced levels of PhIP-DNA adducts as compared with controls given PhIP alone. The possible mechanism of protective effect of tea on PhIP-DNA adduct formation was then examined in vitro. It was found that an aqueous extract of green and black tea, mixtures of green and black tea polyphenols, as well as purified polyphenols could strongly inhibit the DNA binding of N-acetoxy-PhIP, a putative ultimate carcinogen of PhIP formed in vivo via metabolic activation. Among these, epigallocatechin gallate was exceptionally potent. HPLC analyses of these incubation mixtures containing N-acetoxy-PhIP and the tea polyphenols each revealed the production of the parent amine, PhIP, indicating the involvement of a redox mechanism. In view of the presence of relatively high levels of tea polyphenols in rat and human plasma after ingestion of tea, this study suggests that direct reduction of the ultimate carcinogen N-acetoxy-PhIP by tea polyphenols is likely to be involved in the mechanism of chemoprotection of tea against this carcinogen. PMID- 12628518 TI - Cancer chemopreventive activity and bioavailability of tea and tea polyphenols. AB - Consumption of tea (Camellia sinensis) has been associated with many health benefits including the prevention of cancer. Based on in vitro experiments, many mechanisms have been proposed to account for the cancer chemopreventive activity. The importance of some of these mechanisms in vivo remains in question due to an incomplete understanding of the bioavailability of the polyphenolic compounds in tea. In this article, the literature on the cancer chemopreventive activity of tea and the tea polyphenols is discussed as well as some of the possible mechanisms for this activity. Whereas studies in animal models and with cell lines have demonstrated cancer preventive activity, the epidemiological data remain mixed. This discrepancy may arise from several factors including lifestyle, correlation between animal models and humans, and differences in metabolism among individuals. Results on the bioavailability and biotransformation of the tea polyphenols help explain some of the differences. We hope this article will spark research efforts on some of the important questions regarding tea polyphenol bioavailability and cancer chemoprevention. PMID- 12628519 TI - Chemoprevention with chlorophyllin in individuals exposed to dietary aflatoxin. AB - Because of the multiplicative interaction between dietary aflatoxins and hepatitis B virus infection in the etiology of liver cancer, efforts to reduce the consequences of either chemical or viral component are likely to have substantial public health benefit. Chlorophyllin (CHL), a water-soluble form of chlorophyll, was recently evaluated as a chemopreventive agent in a population at high risk for exposure to aflatoxin and subsequent development of hepatocellular carcinoma. CHL, which is used extensively as a food colorant and has numerous medicinal applications, is an effective anticarcinogen in experimental models including aflatoxin-induced hepatocarcinogenesis. CHL is thought to form molecular complexes with carcinogens, thereby blocking their bioavailability. In the clinical trial, administration of CHL three times a day led to a 50% reduction in the median level of urinary excretion of aflatoxin-N(7)-guanine compared to placebo. This excreted DNA adduct biomarker is derived from the ultimate carcinogenic metabolite of aflatoxin B(1), aflatoxin-8,9-epoxide, and is associated with increased risk of developing liver cancer in prospective epidemiologic studies. Compliance in the intervention was outstanding and no toxicities were observed. Thus, CHL has been found to be a safe and effective agent suitable for use in individuals unavoidably exposed to aflatoxins. PMID- 12628521 TI - Measurement of oxidative damage at individual gene levels by quantitative PCR using 8-hydroxyguanine glycosylase (OGG1). AB - In this study, an attempt was made to develop a method to estimate oxidative damage of individual genes for assessing chemopreventive potential of dietary or medicinal plants. Oxidative damage was investigated on the two genes in gastric mucosal tissue infected with Helicobacter pylori, which were genes of glyceraldehydes-3-phosphate dehydrogenase (GAPDH), a house-keeping gene, and gene of insulin-like growth factor II receptor (IGFIIR), a gene known to be mutated frequently in gastric carcinoma. The oxidative damage in genomic DNA in the above tissue was confirmed by immunohistochemical study using monoclonal antibody to 8 hydroxyguanine (oh(8)G), which showed much higher degree of staining in their nuclei. Using the method we developed, it was demonstrated that the number of oh(8)G (indicated by 8-hydroxyguanine glycosylase (OGG1) sensitive sites) in GAPDH was almost not changed in H. pylori-infected tissue but in IGFIIR, it increased significantly. These results indicate that this method is valid for the estimate of oxidative damage of individual genes and also showed that the susceptibility of genomic DNA to attack of reactive oxygen species is not homogeneous but different depending upon the region of DNA. We expect to use this method in studies of carcinogenic mechanism and chemoprevention since it can provide more specific information pertaining to individual genes we are interested in. PMID- 12628520 TI - Promotion versus suppression of rat colon carcinogenesis by chlorophyllin and chlorophyll: modulation of apoptosis, cell proliferation, and beta-catenin/Tcf signaling. AB - The carcinogens 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and 1,2 dimethylhydrazine (DMH) induce colon tumors in the rat that contain mutations in beta-catenin, but the mutation pattern can be influenced by exposure to dietary phytochemicals, such as the water-soluble derivative of chlorophyll called chlorophyllin. Whereas chlorophyllin is an effective blocking agent during the initiation phase, post-initiation responses depend upon the exposure protocol, and can be influenced by the initiating agent and the concentration of chlorophyllin. Post-initiation treatment with 0.001% chlorophyllin (w/v) in the drinking water promoted colon carcinogenesis in the rat, but much higher concentrations (1.0% chlorophyllin) led to suppression. Bromodeoxyuridine and terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) indices revealed that the promotional concentration of 0.001% chlorophyllin increased the ratio of cell proliferation to apoptosis in the colonic crypts, whereas concentrations in the range 0.0l-1.0% chlorophyllin modestly reduced this ratio. Molecular studies showed that the spectrum of beta-catenin mutations was markedly different in chlorophyllin-promoted colon tumors--many of the mutations led to direct substitutions of critical Ser/Thr residues within the glycogen synthase kinase-3beta (GSK-3beta) region, whereas in all other groups, including DMH and IQ controls, the mutations typically affected amino acids adjacent to Ser(33). Substitution of critical Ser/Thr residues caused beta-catenin and c-Jun proteins to be markedly over-expressed compared with tumors in which the mutations substituted amino acid residues flanking these critical Ser/Thr sites. In a separate study, rats were exposed to IQ or azoxymethane (AOM), a metabolite of DMH, and they were treated post-initiation with chlorophyllin, chlorophyll, copper, or phytol in the diet. Natural chlorophyll (0.08%) suppressed AOM- and IQ induced aberrant crypt foci (ACF), whereas chlorophyllin had no effect and copper promoted the number of small ACF induced by IQ. The results suggest that further investigation of the dose-response for suppression versus promotion by chlorophyll and chlorophyllin is warranted, including studies of the beta catenin/Tcf signaling pathway and its influence on cell proliferation and apoptosis in the colonic crypt. PMID- 12628522 TI - Modulation of cigarette smoke-related end-points in mutagenesis and carcinogenesis. AB - The epidemic of lung cancer and the increase of other tumours and chronic degenerative diseases associated with tobacco smoking have represented one of the most dramatic catastrophes of the 20th century. The control of this plague is one of the major challenges of preventive medicine for the next decades. The imperative goal is to refrain from smoking. However, chemoprevention by dietary and/or pharmacological agents provides a complementary strategy, which can be targeted not only to current smokers but also to former smokers and passive smokers. This article summarises the results of studies performed in our laboratories during the last 10 years, and provides new data generated in vitro, in experimental animals and in humans. We compared the ability of 63 putative chemopreventive agents to inhibit the bacterial mutagenicity of mainstream cigarette smoke. Modulation by ethanol and the mechanisms involved were also investigated both in vitro and in vivo. Several studies evaluated the effects of dietary chemopreventive agents towards smoke-related intermediate biomarkers in various cells, tissues and organs of rodents. The investigated end-points included metabolic parameters, adducts to haemoglobin, bulky adducts to nuclear DNA, oxidative DNA damage, adducts to mitochondrial DNA, apoptosis, cytogenetic damage in alveolar macrophages, bone marrow and peripheral blood erytrocytes, proliferation markers, and histopathological alterations. The agents tested in vivo included N-acetyl-L-cysteine, 1,2-dithiole-3-thione, oltipraz, phenethyl isothiocyanate, 5,6-benzoflavone, and sulindac. We started applying multigene expression analysis to chemoprevention research, and postulated that an optimal agent should not excessively alter per se the physiological background of gene expression but should be able to attenuate the alterations produced by cigarette smoke or other carcinogens. We are working to develop an animal model for the induction of lung tumours following exposure to cigarette smoke. The most encouraging results were so far obtained in models using A/J mice and Swiss albino mice. The same smoke-related biomarkers used in animal studies can conveniently be applied to human chemoprevention studies. We participated in trials evaluating the effects of N-acetyl-L-cysteine and oltipraz in smokers from Italy, The Netherlands, and the People's Republic of China. We are trying to develop a pharmacogenomic approach, e.g. based on genetic metabolic polymorphisms, aimed at predicting not only the risk of developing cancer but also the individual responsiveness to chemopreventive agents. PMID- 12628523 TI - Chemoprevention of mammary carcinogenesis by 1alpha-hydroxyvitamin D5, a synthetic analog of Vitamin D. AB - Numerous analogs of Vitamin D have been synthesized in recent years with the hope of generating a compound that retains the anticarcinogenic activity of Vitamin D without causing any toxicity. We synthesized such an analog, 1alpha-hydroxy-24 ethylcholecalciferol [1alpha-hydroxyvitamin D(5) or 1alpha(OH)D(5)], and showed that it was tolerated by rats and mice at a much higher dose than 1alpha,25 dihydroxy cholecalciferol [1alpha,25(OH)(2)D(3)]. This property makes it a prime candidate for chemoprevention studies. In the mouse mammary gland organ culture (MMOC), 1alpha(OH)D(5) inhibited carcinogen-induced development of both mammary alveolar and ductal lesions. In vivo carcinogenesis study showed statistically significant reduction of tumor incidence and multiplicity in N-methyl-N nitrosourea (MNU)-treated rats that were fed 25-50 microg 1alpha(OH)D(5)/kg diet. There were no adverse effects on plasma calcium concentrations. In order to determine if the effect of 1alpha(OH)D(5) would be selective in suppressing proliferation of transformed cells, its effects on cell growth and proliferation were compared between BT474 (cancer) and MCF12F (non-tumorigenic) human breast epithelial cells. Results showed that 1alpha(OH)D(5) induced apoptosis and cell cycle G1 phase arrest in BT474 breast cancer cells without having any effects on proliferation of the MCF12F cells. In addition, in MMOC it had no growth inhibitory effects on normal epithelial cell proliferation in the absence of carcinogen. Similarly, non-tumorigenic human breast epithelial cells in explant culture did not respond to 1alpha(OH)D(5), whereas treatment with 1alpha(OH)D(5) induced cell death in the explants of cancer tissue. These results collectively indicate that 1alpha(OH)D(5) selectively induced apoptosis only in transformed cells but not in normal breast epithelial cells. Interestingly, the growth inhibitory effects of 1alpha(OH)D(5) were observed in Vitamin D receptor positive (VDR(+)) breast cancer cells, but not in highly metastatic VDR(-) breast cancer cells, such as MDA-MB-435 and MDA-MB-231, suggesting that 1alpha(OH)D(5) action may be mediated, in part, by VDR. PMID- 12628524 TI - Beneficial and adverse effects of chemopreventive agents. AB - The beneficial and adverse effects of some chemopreventive agents, such as Vitamins A, C, E, beta-carotene, indole-3-carbinol, capsaicin, garlic, and aloe are reviewed. Two large randomized trials with a lung cancer endpoint, the Alpha Tocopherol, Beta-Carotene (ATBC) Prevention Study and the Beta-Carotene and Retinol Efficacy Trial (CARET), suggested that antioxidants might be harmful in smokers. However, the results of the Linxian study and of the ATBC or the CARET studies were significantly different in this respect, and therefore, the relationship between antioxidant and carcinogenesis remains open to debate. Indole-3-carbinol has cancer promoting activities in the colon, thyroid, pancreas, and liver, whereas capsaicin alters the metabolism of chemical carcinogens and may promote carcinogenesis at high doses. Organosulfur compounds and selenium from garlic have no or a little enhancing effect on cancer promotion stage. Information upon chemopreventive mechanisms that inhibit carcinogenesis is imperfect, although the causes and natures of certain human cancers are known. Therefore, definitive preventive guidelines should be carefully offered for various types of tumors, which properly consider ethnic variations, and the efficacies and the safety of chemopreventive agents. PMID- 12628525 TI - Tuberculosis and pregnancy. AB - OBJECTIVES: There are many myths surrounding pregnancy and tuberculosis (TB), and outcome of treatment. This prospective study was conducted at the Department of Obstetrics and Gynecology, SCB Medical College, Cuttack, India, and at the authors' private clinics from 1986 to 2001 to determine the outcome of pregnancy if TB is treated properly. METHODS: A total of 111 pregnant women diagnosed as having pulmonary and glandular TB were included in the study. They were matched for age, parity, and socioeconomic status with 51 pregnant women without TB (first control group), and 51 women with pulmonary TB but without pregnancy (second control group). The usual pregnancy management was given to the women in the study group, along with a short course of chemotherapy: either ethambutol, INH, or rifampicin and pyrazinamide for 2 months followed by INH and rifampicin for 4 months; or ethambutol, INH, and rifampicin for 2 months followed by INH and rifampicin for 7 months. Statistical analysis was done using a chi(2)-test. RESULTS: There were no statistical differences in duration of gestation, preterm labor, and other complications of pregnancy, labor, and puerperium between the pregnancy groups. There were no congenital anomalies in the babies born to the groups. Pregnancy had no effect on the course of TB as regards sputum conversion, stabilization of the disease, and non-relapse even after 2-5 years of follow-up and a further delivery in a few cases. CONCLUSIONS: If proper and adequate chemotherapy is given to pregnant women with TB, they are not a higher risk than non-pregnant women with TB. Neither the disease nor chemotherapy is threatening to mother or newborn. However, today the ominous combination of human immunodeficiency virus, TB, and pregnancy poses a new challenge. PMID- 12628526 TI - Effect of prolongation of pregnancy on perinatal mortality. AB - OBJECTIVES: (1) To determine the distribution of singleton and twin births according to gestational age in a Nigerian obstetric population; and (2) to compare their perinatal outcomes according to gestational age. METHODS: A 10-year retrospective comparative study of twin and singleton births at a tertiary care center in Enugu, Nigeria. The variables analyzed were: the proportion of deliveries occurring at each gestational age, the gestational age-specific rates for stillbirths, cesarean section, babies with 1-min Apgar scores less than 4 and those whose birthweights were below the 10th percentile for gestational age. The trends in these rates were determined by finding the best fitting polynomial regression curve for each variable. Tests of statistical significance for trends in proportions were carried out by means of the chi(2)-test at the 95% confidence level. RESULTS: Of the 496 twin births, 3.6% compared with 17.3% of the 496 singleton births went beyond 40 weeks' gestation while 1.2% of the twin and 4.4% of the singleton deliveries occurred at 42 weeks' gestation or beyond. For twins as well as singletons, there was a consistent and significant decline in the stillbirth rate and the proportion of babies with 1-min Apgar scores less than 4 up to 42 weeks (P=0.0000). Among the twins, the proportion of babies with birthweights below the 10th percentile (i.e. those with impaired growth) significantly rose from 28 weeks and above (P=0.0000) while among the singletons, a declining trend with gestational age was observed (P=0.0003). However, among the twins with impaired growth, the stillbirth rate neither differed between the first and second twins at each gestational age nor did it increase with gestational age in both the first and second twins. While the cesarean section rate for singletons remained almost stable at approximately 13%, there was a significant rise in the cesarean section rate with gestational age among the twin births. CONCLUSIONS: There were 1.2% of twin deliveries compared with 4.4% of singleton deliveries which occurred at 42 weeks' gestation or beyond. In the Nigerian population studied, the perinatal outcomes in twins did not differ from those of singletons up to 42 weeks' gestation suggesting that the 42-week cut-off for prolonged pregnancy applies equally well to twins as to singletons. PMID- 12628527 TI - Recrudescent herpes labialis during and prior to early pregnancy. AB - OBJECTIVES: To assess the experience of recrudescent herpes labialis (RHL) before and during early pregnancy. METHODS: History of RHL prior to and during the first trimester of pregnancy was obtained from 3738 women attending at 10-15 weeks' gestation. The influence of age, ethnicity, socioeconomic group, smoking behavior, and alcohol intake on RHL was assessed. RESULTS: 1066 women (28.5%) reported a history of RHL lesions, with reduced incidence of RHL during pregnancy (0.111 lesions/subject per month) compared with outside pregnancy (0.19 lesions/subject per month) (P<0.0001). Those who did report lesions during pregnancy (n=296) experienced them at a higher monthly rate (0.41 lesions/subject per month) than before pregnancy (0.25 lesions/subject per month) (P<0.0001). RHL rate in early pregnancy was related solely to the previous rate of lesion recrudescence (P<0.001). CONCLUSION: Pregnant women with a history of RHL report reduced incidence of RHL during pregnancy. PMID- 12628528 TI - A comparison of two methods of labor induction with vaginal misoprostol. AB - OBJECTIVES: To evaluate if labor induction with 50 microg of vaginal misoprostol twice per day is as effective as and safer than 100 microg used once per day. METHODS: Misoprostol was used to induce labor in 204 consecutive pregnant women assessed as needing labor induction, 104 at the University Hospital of the West Indies in Kingston, Jamaica, and 100 at the Victoria Jubilee Hospital, also in Kingston. At the former institution the women were administered 100 microg of misoprostol once per 24 h and at the latter 50 microg every 12 h. The doses were repeated if there was no cervical change or if the woman was not in labor. Bishop scores were determined before initiation of induction and again 12 h later. Women's records were reviewed after 24 h to determine delivery outcome. RESULTS: The indications for labor induction were similar in each group. There was no significant difference in the group demographics. In the group given 50 microg of misoprostol twice per day the mean time+/-S.D. from insertion of misoprostol to delivery was significantly shorter than in the other group (560.14+/-269.20 min vs. 729.90+/-471.65 min; P<0.01), and the percentage of women who were delivered within 12 h was higher (75% vs. 56.8%; P<0.002). There was no significant difference between the two groups in the rate of cesarean births, the need for oxytocin, or blood loss. Only two patients experienced uterine hyperstimulation, both in the group that had received 100 microg of misoprostol per day. There was no significant difference between the two groups in the birthweight of the neonates and in the number of neonates with Apgar scores less than 7 at 1 min and 5 min. There were, however, significantly fewer neonates who needed resuscitation (6% vs. 14.7%; P=0.04) and admission to the special care nursery (7% vs. 17%; P=0.03) in the group that had received 50 microg of misoprostol twice per day. CONCLUSIONS: A dose of 50 microg of misoprostol twice per day appears to be more efficient and safer than a dose of 100 microg once per day, but this may partially be due to weaknesses in the study design. PMID- 12628529 TI - Calcium to creatinine ratio in a spot sample of urine for early prediction of pre eclampsia. AB - OBJECTIVES: Based on the fact that urinary calcium excretion decreases in pre eclampsia, this study was designed to determine the predictive value of calcium to creatinine ratio in a spot urine sample. METHODS: The calcium to creatinine ratio was measured in a spot urine sample of 102 normotensive women at 20-24 weeks' gestation who attended the prenatal care clinic of the Shiraz University of Medical Sciences. The women were followed-up until delivery and grouped according to pre-eclampsia occurrence. The prevalence of pre-eclampsia was measured and compared with the calcium to creatinine ratio. RESULTS: Ninety-four women remained normotensive during pregnancy and eight developed pre-eclampsia. Mean age, gestational age at the beginning of the study, and gestational age at delivery did not differ significantly between the two groups. Mean urinary calcium concentration (15.9+/-8.5 mg/dl in normotensive vs. 10.2+/-7.5 mg/dl in pre-eclamptic women), and mean birth weight (3192+/-336.3 g vs. 2712+/-468.9 g) were significantly lower in pre-eclamptic patients (P=0.03 and 0.005, respectively). Mean calcium to creatinine ratio was also significantly lower in the pre-eclamptic group (P<0.03). CONCLUSIONS: Single urine calcium to creatinine ratio may be an effective method for screening women at greatest risk for pre eclampsia. PMID- 12628530 TI - Ultrasonographic patterns of fetal breathing movements in normal pregnancy. AB - OBJECTIVE: To assess through pregnancy fetal breathing movements (FBMs) patterns detected by M-mode and Doppler velocimetry technology. METHODS: In this cross sectional study FBMs were investigated in 1882 uncomplicated pregnancies over a 4 year period. Abdominal and thoracic wall movements of fetuses between 14 and 40 weeks of gestation were studied by M-Mode scan, and color Doppler velocimetry with spectral imaging analysis was used to investigate the presence of FBMs associated with nasal fluid flow velocity waveforms (NFFVWs). RESULTS: Abdominal movements were observed in 19% of cases when gestation was less than 20 weeks and in 61% of cases when it was between 21 and 25 weeks; chest movements were significant after 21 weeks; and NFFVWs were detected at 22 weeks and increased progressively to 93% of cases at term. CONCLUSIONS: Fetal breathing movements are a complex phenomenon with a composite, progressive pattern of development during gestation. PMID- 12628531 TI - Dexamethasone compared with betamethasone for glucocorticoid treatment of postpartum HELLP syndrome. AB - OBJECTIVES: To compare the efficacy of dexamethasone and betamethasone to ameliorate the course of postpartum hemolysis, elevated liver enzymes, low platelets (HELLP) syndrome. METHODS: A prospective, mixed randomized/non randomized clinical investigation of patients with postpartum HELLP syndrome. Treatment with either dexamethasone or betamethasone was continued until there was evidence of disease recovery. RESULTS: Baseline characteristics of both the dexamethasone (n=18) and betamethasone (n=18) groups were similar. Although the time to discharge from the obstetrical recovery room was not statistically significant between groups, reduction in mean arterial blood pressure was more pronounced in the dexamethasone group as compared with the betamethasone group ( 15.3+/-1.4 mmHg vs. -7.5+/-1.4 mmHg, respectively, P<0.01). Patients in the dexamethasone group required less antihypertensive treatment than the betamethasone group (6% vs. 50%, P=0.01) and also had a decreased need for readmission to the obstetrical recovery room (0% vs. 22%, P=0.03). CONCLUSION: This investigation supports the use of dexamethasone as the superior glucocorticoid to use for patients with postpartum HELLP syndrome. PMID- 12628532 TI - Transvaginal color Doppler ultrasonography for prediction of pre-cancerous endometrial lesions. AB - OBJECTIVE: To determine whether measurements of blood flow in endometrial and uterine vessels by transvaginal color Doppler ultrasonography was valuable in the diagnosis of a neoplastic endometrial pathology (hyperplasia and carcinoma) in women with abnormal bleeding. METHODS: This is a prospective study and included 105 post-menopausal women and 33 pre-menopausal women with abnormal uterine bleeding. All subjects underwent transvaginal color Doppler ultrasonography. We investigated whether obtained results were correlated with histopathological findings. RESULTS: There was no significant difference in the mean+/-S.D. RI of the left and the right uterine arteries, intramyometrial arteries and endometrial arteries between patients with neoplastic and non-neoplastic endometrium on histopathological examination. Doppler's velocity waveforms of small endometrial blood vessels could be detected in 9% of the women with non-neoplastic endometrium and in 42% of the women with neoplastic endometrium (P<0.05). The mean+/-S.D. of the endometrial thickness was significantly higher in the women with neoplastic endometrium than that of the women with non-neoplastic endometrium (16.6+/-6.1 mm vs. 9.5+/-4.7 mm, P<0.05). CONCLUSION: Doppler's velocity waveforms of uterine vessels coupled with transvaginal ultrasonography are not valuable enough to replace histopathological examination in the diagnosis of a neoplastic endometrial pathology. However, it may be helpful in cases in which invasive techniques are difficult to perform and in the differentiation of a certain group of patients at little risk of endometrial carcinoma. PMID- 12628533 TI - Endometrial thickness and histopathologic findings in women with postmenopausal bleeding. PMID- 12628534 TI - Successful medical management of ovarian pregnancy. PMID- 12628535 TI - Menstrual-reproductive factors and age at natural menopause in Iran. PMID- 12628536 TI - Histologic examination of products of conception at the time of pregnancy termination. PMID- 12628537 TI - Second-trimester pregnancy termination with misoprostol in women with previous cesarean sections. PMID- 12628538 TI - Gestational age-specific neonatal mortality among preterm singleton births in a Nigerian tertiary institution. PMID- 12628539 TI - Acoustic neuroma in pregnancy: emergency cesarean section and definitive neurosurgery. PMID- 12628540 TI - Primary omental pregnancy treated by laparoscopic surgery. PMID- 12628541 TI - Classification of perinatal death in a developing country. AB - OBJECTIVES: To evaluate and compare the three most commonly used perinatal death classification systems: (1) the Nordic-Baltic; (2) the Aberdeen; and (3) the Wigglesworth, and assess their applicability in a developing country (Sudan) with a high perinatal mortality rate, and their justification for practical use in quality assurance and audit activities. METHODS: At Omdurman Maternity Hospital (OMH), Khartoum, Sudan, 166 perinatal deaths were prospectively assessed during a 3-month period (May-August 2000) with a total of 2260 births. Narratives of 166 perinatal deaths were prepared for the purpose of audit. A panel of two Danish and one Sudanese obstetrician categorized the cases according to: (1) the Nordic Baltic; (2) the Aberdeen; and (3) the Wigglesworth classification. RESULTS: By all three classifications a similar fraction of cases (approx. 85%) were allocated to one category only, and in 15% of cases the assessors were in doubt into which of two categories the cases should be allocated. The necessary information is often not available, giving at least 40% classified as 'unknown' in the Aberdeen classification, whereas the Wigglesworth classification results in an even larger group of unspecified asphyxia. CONCLUSION: Classification of perinatal deaths in developing countries is associated with problems regarding application, validity and usefulness. The Nordic-Baltic classification seems to be most suitable for appropriate stratification using routinely recorded variables and providing categories associated with specific levels of care. PMID- 12628542 TI - Lack of correlation between O-serotype, bacteriophage susceptibility and genomovar status in the Burkholderia cepacia complex. AB - The Burkholderia cepacia complex comprises at least nine phylogenetically related genomic species (genomovars) which cause life-threatening infection in immunocompromised humans, particularly individuals with cystic fibrosis or chronic granulomatous disease. Prior to recognition that 'B. cepacia' comprise multiple species, in vitro studies revealed that the lipopolysaccharide (LPS) of these Gram-negative bacteria is strongly endotoxic. In this study, we used 117 B. cepacia complex isolates to determine if there is a correlation between O-antigen serotype and genomovar status. Isolates were also tested for their ability to act as bacterial hosts for the LPS-binding bacteriophages NS1 and NS2. The absence of genomovar II (Burkholderia multivorans) in 'historical B. cepacia' isolates was notable. Neither O-serotype nor phage susceptibility correlated with genomovar status. We conclude that variability in LPS may contribute to the success of these highly adaptable bacteria as human pathogens. PMID- 12628543 TI - K-antigen-specific, but not O-antigen-specific natural human serum antibodies promote phagocytosis of Klebsiella pneumoniae. AB - Infections due to Klebsiella pneumoniae and other Klebsiella spp. are a leading cause of hospital-associated morbidity, especially in the intensive care setting. In this study, the hypothesis that normal human sera contain sufficient concentrations of K-antigen-specific antibodies to promote phagocytic killing of encapsulated, highly virulent Klebsiella organisms was tested. K2-antigen specific IgG and IgM antibodies were detected in each of 10 normal sera, and such antibodies were functionally active in a phagocytic killing assay. Phagocytosis depended critically on sufficient numbers of neutrophils and was impaired by the presence of soluble Klebsiella capsular polysaccharide (CPS). Thus, insufficient numbers of neutrophils and circulation of soluble CPS but not lack of K-specific antibodies may be detrimental in Klebsiella sepsis. The efficacy of hyperimmune sera might be based not on enhancement of phagocytosis but on the neutralization of these detrimental effects of circulating CPS and LPS. PMID- 12628544 TI - Staphylococcus aureus strains isolated from bovine mastitis: virulence, antibody production and protection from challenge in a mouse model. AB - Septic arthritis in mice was used as a model to evaluate the virulence of Staphylococcus aureus and coagulase-negative staphylococci (CNS) isolated from cases of bovine mastitis. In addition, the model was used to evaluate the cross protection elicited by heterologous antibodies. Mice were intramuscularly inoculated with serial bacterial doses of different strains of S. aureus or CNS, for virulence determination; they were monitored for arthritis, gangrene or death up to 20 days. Antibody response, cross reactivity and resistance to challenge were tested by subcutaneous inoculation with a low dose of one of the S. aureus or CNS strains followed by challenge with two S. aureus strains. S. aureus alpha hemolysin isolate was the most virulent, followed by alpha+beta-hemolysin and beta-hemolysin isolates. The least virulent isolates were the non-hemolytic S. aureus strains but even they were more virulent than the CNS strains tested. Antibodies against three different S. aureus antigens were detected by the ELISA in all mice that were inoculated with the S. aureus strains but not in any of those with the CNS strains. Immunoblot test against various S. aureus strains as antigens showed high cross-reactivity among the S. aureus strains but only a slight similarity, restricted to the bands above 36 kDa, with the CNS sera. Low dose inoculation of alpha or alpha+beta strains before challenge with homologous and heterologous strains protected the mice, whereas the two beta strains provided only partial protection. The inoculations of non-hemolytic S. aureus or the CNS strains did not elicit any protection. Our findings demonstrate that pre exposure of mice to a low dose of certain S. aureus strains could provide protection and that the antibodies produced could have an important protective role. PMID- 12628545 TI - Anti-Chlamydia pneumoniae heat shock protein 10 antibodies in asthmatic adults. AB - A multicenter prospective study was performed on 160 asthmatic adults suffering from acute episodes of bronchitis and 88 non-asthmatic controls, to investigate potential associations among Chlamydia pneumoniae infection and/or anti-C. pneumoniae heat shock protein 10 antibodies, and asthma. We used micro immunofluorescence to detect serum anti-C. pneumoniae IgG, IgA and IgM antibodies and enzyme-linked immunosorbent assay to detect serum anti-Chsp10 peptide IgG antibodies. The serological prevalence of C. pneumoniae was 73.1%. An association was observed between the presence of anti-Chsp10 antibodies and adult onset asthma. The humoral immune responses were not confined to any particular region of the Chsp10 protein. PMID- 12628547 TI - Correlation between CMV genotypes, multiple infections with herpesviruses (HHV-6, 7) and development of CMV disease in kidney recipients in Kuwait. AB - The possible correlation between cytomegalovirus, human herpesvirus types 6, 7 and cytomegalovirus-related clinical symptoms was studied in kidney transplant patients in Kuwait. Cytomegalovirus infection was diagnosed using the pp65 antigenemia assay. DNA of cytomegalovirus was detected by nested polymerase chain reaction (nested-PCR). PCR was also used to amplify the genes coding for structural proteins of human herpesvirus-6 (240 bp) and human herpesvirus-7 (186 bp). Glycoprotein B genotypes of cytomegalovirus were determined by restriction fragment length polymorphism. The average number of cells positive for cytomegalovirus pp65 antigen showed a steady increase with the severity of the cytomegalovirus-related symptoms. Furthermore, cytomegalovirus pp65 antigen positivity was significantly more frequent among recipients of cadaver kidney (45.5%) than among those who received live related kidneys (22.6%). Cytomegalovirus gB genotype 1 was detected more frequently (P<0.036) in recipients with live related donor kidney (38%) than in patients of cadaver kidney (13%). The genome of human herpesvirus-6 was detected at the same rate in patients with or without cytomegalovirus-related symptoms. However, the genome of human herpesvirus-7 was detected significantly more frequently (P<0.0001) in asymptomatic patients (41.7%) than in recipients with symptomatic cytomegalovirus infection (17%). We conclude that cytomegalovirus gB genotypes are not associated with the outcome of a cytomegalovirus infection in kidney transplant patients, that human herpesvirus-6 does not play a role in cytomegalovirus pathogenesis and that the role of human herpesvirus-7 in cytomegalovirus-related morbidity in kidney recipients remains unclear. PMID- 12628546 TI - The role of surface carbohydrates on the interaction of microconidia of Trichophyton mentagrophytes with epithelial cells. AB - The presence of carbohydrate-binding adhesins on the microconidia of Trichophyton mentagrophytes surface and their role on cellular interactions were investigated. Flow cytometry showed that this fungus recognizes the sugars mannose and galactose. The binding was inhibited by the addition of methyl alpha-D mannopyranoside and methyl alpha-D-galactopyranoside, and showed higher fluorescence intensity at 37 degrees C than 28 degrees C. Trypsin treatment and heating of the cells reduced the binding, suggesting a (glyco) protein nature of the microconidia adhesin. The interaction of the fungus to Chinese hamster ovary epithelial cells and its glycosylation-deficient mutants demonstrated a higher adhesion index in Lec1 and Lec2 mutants, which express mannose and galactose, respectively, as the terminal carbohydrate on the cell surface. Endocytosed fungi were shown preferentially in Lec2 cells. Addition of the carbohydrates methyl alpha-D-mannopyranoside and methyl alpha-D-galactopyranoside to the interaction medium, pretreatment of Lec1 and Lec2 cells with lectins Concanavalina A and Arachis hypogaea and pretreatment with sodium periodate decreased the adhesion and the endocytic index. Examination of thin section by transmission electron microscopy showed that after fungal ingestion by Lec2 cells the fungi are enclosed in a 'loose'-type vacuole while the other cells are found within a 'tight'-type membrane-bound cytoplasmic vacuole. Our results suggest the occurrence of carbohydrate-specific adhesins on microconidia surface that recognize mannose and galactose. This may have a role in the adhesion process during the infectious process of dermatophytosis. PMID- 12628549 TI - Lack of biofilm contribution to bacterial colonisation in an experimental model of foreign body infection by Staphylococcus aureus and Staphylococcus epidermidis. AB - The contribution of in vivo biofilm-forming potential of Staphylococcus aureus and Staphylococcus epidermidis was studied in an experimental model of foreign body infections. Increasing inocula (from 10(2) to 10(7) organisms) of ica positive strains of S. aureus and S. epidermidis and their ica-negative isogenic mutants (the ica locus codes for a major polysaccharide component of biofilm) were injected into subcutaneously implanted tissue cages in guinea pigs. Surprisingly, bacterial counts and time-course of tissue cage infection by ica positive strains of S. aureus or S. epidermidis were equivalent to those of their respective ica-negative mutants, in the locally infected fluids and on tissue cage-inserted plastic coverslips. PMID- 12628548 TI - Oral use of Lactobacillus rhamnosus GR-1 and L. fermentum RC-14 significantly alters vaginal flora: randomized, placebo-controlled trial in 64 healthy women. AB - Urogenital infections afflict an estimated one billion people each year. The size of this problem and the increased prevalence of multi-drug resistant pathogens make it imperative that alternative remedies be found. A randomized, placebo controlled trial of 64 healthy women given daily oral capsules of Lactobacillus rhamnosus GR-1 and Lactobacillus fermentum RC-14 for 60 days showed no adverse effects. Microscopy analysis showed restoration from asymptomatic bacterial vaginosis microflora to a normal lactobacilli colonized microflora in 37% women during lactobacilli treatment compared to 13% on placebo (P=0.02). Lactobacilli were detected in more women in the lactobacilli-treated group than in the placebo group at 28 day (P=0.08) and 60 day (P=0.05) test points. Culture findings confirmed a significant increase in vaginal lactobacilli at day 28 and 60, a significant depletion in yeast at day 28 and a significant reduction in coliforms at day 28, 60 and 90 for lactobacilli-treated subjects versus controls. The combination of probiotic L. rhamnosus GR-1 and L. fermentum RC-14 is not only safe for daily use in healthy women, but it can reduce colonization of the vagina by potential pathogenic bacteria and yeast. PMID- 12628550 TI - N-terminus of M2 protein could induce antibodies with inhibitory activity against influenza virus replication. AB - New influenza vaccines have been designed based on the fact that the extracellular domain of M2 protein (M2e) is nearly invariant in all influenza A strains. To clarify which exact region of M2e could induce antibodies with inhibitory activities against influenza virus replication, four overlapping peptides covering M2e were synthesized and then coupled to the carrier protein bovine serum albumin through the cysteine of the peptides. After a vaccination course, all these four peptide vaccines could induce high levels of rabbit antibodies with predefined peptide specificity (antibody dilution: 1:6400 1:25600). Besides, the anti-N-terminal antibodies (AS2) reacted strongly with M2e, and reacted weakly with the middle part and C-terminus of M2e. The MDCK assay for cytopathic effect proved that antibodies recognizing the N-terminus of M2e could obviously inhibit replication of influenza A virus (A/wuhan/359/95) and influenza B virus (B/wuhan/321/99) in vitro in a dose-dependent manner, while antibodies recognizing the middle part and the C-terminus of M2e did not show such significant inhibitory activities. Sequence analysis indicates that the first nine N-terminal amino acid residues of M2e are extremely conservative. Just this region containing the first nine amino acid residues could induce antibodies with inhibitory activity against influenza A and influenza B virus replication, suggesting that the N-terminus of M2e may contain an epitope that could induce inhibitory antibodies against influenza virus replication in vitro. PMID- 12628551 TI - Flow cytometric measurements of neutrophil functions: the dependence on the stimulus to cell ratio. AB - Phagocytosis and antimicrobial killing of neutrophils has been quantitatively determined as a function of the stimulus (Candida albicans) to cell ratio R using two donor collectives containing a total of 115 blood samples. Analysis of the collectives in two different laboratories according to the same flow cytometric protocol for simultaneous measurement of neutrophil functions did not produce statistically significant differences. The number of phagocytosing leukocytes as well as that of killed fungi per leukocyte depends strongly on R. While each phagocytosing neutrophil kills one fungus at low values of R, each neutrophil kills on average 2.5 fungi for large R. PMID- 12628552 TI - TT virus in Hungary: sequence heterogeneity and mixed infections. AB - The majority of the viral hepatitis cases is caused by five hepatitis viruses (A,B,C,D,E). In 1997, TT virus was discovered. It was supposed that a number of the unknown hepatitis cases was caused by the TT virus. The aim of this study was to characterize TT viruses carried by healthy individuals and patients suffering from hepatitis of unknown origin in Hungary. TTV DNA was detected by seminested PCR with the commonly used N22 primers. Twenty of the 108 sera (18.5%) taken from healthy persons and 115 of the 228 sera (50.4%) of patients with hepatitis of unknown origin were found to be positive. The nucleotide sequences of 26 clones derived from 17 hepatitis patients and 15 clones from nine healthy persons were determined and a phylogenetic tree was constructed. Genotype 2 (group 1) was found to be the most frequent, but other group 1 genotypes (1, 6) and genotypes 8 and 17 of group 2 were also detected. Mixed TTV infections were found in eight cases (two healthy persons and six hepatitis patients). Variants belonging to the same group were carried in seven cases, and the presence of group 1 (genotype 2) and group 2 (genotype 8) TTV sequences were found in one single hepatitis patient. PMID- 12628553 TI - Impaired neutrophil function in patients with pulmonary tuberculosis and its normalization in those undergoing specific treatment, except the HIV-coinfected cases. AB - Our study investigated whether the respiratory burst (RB) of polymorphonuclear neutrophils from tuberculosis (TB) patients was related with the disease severity or treatment, as well as the circulating levels of TNF-alpha. The sample comprised 57 patients with moderate (n=21) or advanced disease (n=36, 13 of them with HIV coinfection, TB-HIV) and 12 controls. Patients were newly diagnosed (n=27) or under treatment (moderate=14, advanced=10, TB-HIV=6). Cytometric analysis showed that untreated patients had a depressed RB in response to Candida albicans, being more pronounced in the advanced group and nearly absent in TB-HIV cases. A recovered RB was observed in treated patients, except for the TB-HIV cases that continued to show a poor response. TNF-alpha serum levels were increased in untreated patients, mostly in the advanced and TB-HIV groups, and showed an inverse and significant correlation with the RB. Disease severity and anti-TB therapy exerted negative and positive influences on the reactive oxygen intermediates production, respectively. PMID- 12628554 TI - Serologic evidence of human infection by Francisella tularensis in the population of Castilla y Leon (Spain) prior to 1997. AB - Prior to an outbreak in Castilla y Leon in December 1997, tularaemia was practically non-existent in Spain. In this paper we studied the prevalence of antibodies against Francisella tularensis in a representative sample of the population (4825 people) from Castilla y Leon (Spain) in samples collected before this outbreak. Antibodies against F. tularensis were detected in nine (0.19%) of the 4825 sera, with antibody titres ranging from 1/20 to 1/160. Of these nine sera, one was positive in seroagglutination against Brucella. Seroagglutination against other bacteria (Yersinia enterocolitica O:9 and O:3 and Proteus OX19) was negative in all sera. Seroprevalence of antibodies in females was 0.20% and 0.17% in males; no statistically significant differences were found in prevalence in terms of sex, age or province. PMID- 12628555 TI - The resuscitation greats: Eugene Nagel and the Miami paramedic program. PMID- 12628556 TI - In-hospital factors associated with improved outcome after out-of-hospital cardiac arrest. A comparison between four regions in Norway. AB - INTRODUCTION: While pre-hospital factors related to outcome after out-of-hospital cardiac arrest (OHCA) are well known, little is known about possible in-hospitals factors related to outcome. HYPOTHESIS: Some in-hospital factors are associated with outcome in terms of survival. MATERIAL AND METHODS: An historical cohort observational study of all patients admitted to hospital with a spontaneous circulation after OHCA due to a cardiac cause in four different regions in Norway 1995-1999: Oslo, Akershus, Ostfold and Stavanger. RESULTS: In Oslo, Akershus, Ostfold and Stavanger 98, 84, 91 and 186 patients were included, respectively. Hospital mortality was higher in Oslo (66%) and Akershus (64%) than in Ostfold (56%) and Stavanger (44%), P=0.002. By multivariate analysis the following pre arrest and pre-hospital factors were associated with in-hospital survival: age -3.5 mmol l(-1), body temperature T transitions resulting in silent mutations were detected in three individual breast cancer samples. One breast cancer sample also contained a G --> T transversion (Q253H). The SUM-52PE cell line contained an A --> T transversion (AAG --> TAG) resulting in a nonsense mutation (K180Stop). All mutations identified in genomic DNA isolates were in highly conserved regions of the C/EBPD gene. This study indicates that mutational alterations in the coding region of the C/EBPD gene are relatively uncommon in human breast cancer. PMID- 12628589 TI - A rapid method for detecting mutations of the human LDL receptor gene by complete cDNA sequencing. AB - We have developed and clinically tested a rapid and largely automated procedure to detect mutations in the coding region of a gene of interest. Our method relies on the automated sequencing of the complete cDNA, followed by an advanced mutation search-and-verification routine using an integrated set of computer analysis tools. We have applied our automated procedure to the diagnosis of familial hypercholesterolemia (FH) in 52 unrelated FH families, by sequencing the whole cDNA coding region of the LDLR gene. Here we report the procedures and performance of our method in the identification of the most common types of LDLR mutations: short deletions or insertions and point mutations. Our method can provide a standard procedure for the 'overnight' unequivocal identification of mutations in those genetic diseases where several different mutations, none clearly prominent, may affect a given gene. PMID- 12628590 TI - Two novel single nucleotide polymorphisms in the promoter of the cellular retinoic acid binding protein II gene (CRABP-II). AB - The cellular retinoic acid binding protein-II (CRABP-II) is an intracellular protein involved in the transmission of the vitamin A-derived signal which regulates genes responsible for lipid metabolism and adipocyte differentiation. Cellular Retinoic Acid Binding Protein-II gene (CRABP-II) (GDB 134819) is located on chromosome 1q21-23 and this region has been linked with related disorders such as Familial Combined Hyperlipidemia (FCHL), type 2 Diabetes Mellitus, and Lipodystrophy. In this context we hypothesized that CRABP-II is an interesting protein and aimed to provide genetic markers for future studies. In order to do that, we screened the promoter and the entire coding regions for mutations in 53 patients diagnosed with FCHL and 89 normolipidemic controls. Two new single nucleotide polymorphisms (SNPs) were identified in the promoter region a C to A change at position -515 and a T to C substitution at position -394, the latter creating a binding site for SP1. The change -515C > A was identified in a FCHL patient whereas the -394T > C was found in 3 FCHL patients and 4 normolipidemic subjects. This report provides two new polymorphisms in CRABP-II, which can be used as genetic markers for future studies of association or linkage with diseases, particularly those associated with the metabolic syndrome. PMID- 12628591 TI - Detection of type III secretion genes as a general indicator of bacterial virulence. AB - Type III secretion systems of Gram-negative bacteria are specific export machineries for virulence factors which allow their translocation to eukaryotic cells. Since they correlate with bacterial pathogenicity, their presence is used as a general indicator of bacterial virulence. By comparing the genetic relationship of the major type III secretion systems we found the family of genes encoding the inner-membrane channel proteins represented by the Yersinia enterocolitica lcrD (synonym yscV) and its homologous genes from other species an ideal component for establishing a general detection approach for type III secretion systems. Based on the genes of the lcrD family we developed gene probes for Gram-negative human, animal and plant pathogens. The probes comprise lcrD from Y. enterocolitica, sepA from enteropathogenic Escherichia coli, invA from Salmonella typhimurium, mxiA from Shigella sonnei, as well as hrcV from Erwinia amylovora. In addition we included as a control probe the flhA gene from E. coli K-12 to validate our approach. FlhA is part of the flagellar export apparatus which shows a high degree of similarity with type III secretions systems, but is not involved in pathogenicity. The probes were evaluated by screening a series of pathogenic as well as non-pathogenic bacteria. The probes detected type III secretion in pathogens where such systems were either known or were expected to be present, whereas no positive hybridization signals could be found in non pathogenic Gram-negative bacteria. Gram-positive bacteria were devoid of known type III secretion systems. No interference due to the genetic similarity between the type III secretion system and the flagellar export apparatus was observed. However, potential type III secretion systems could be detected in bacteria where no such systems have been described yet. The presented approach provides therefore a useful tool for the assessment of the virulence potential of bacterial isolates of human, animal and plant origin. Moreover, it is a powerful means for a first safety assessment of poorly characterized strains intended to be used in biotechnological applications. PMID- 12628592 TI - First complete large subunit ribosomal RNA sequence and secondary structure for a parasitic nematode: phylogenetic and diagnostic implications. AB - The complete sequence and secondary structure of the large subunit of nuclear ribosomal RNA(LSUrRNA) were determined for the parasitic nematode Labiostrongylus bipapillosus (order Strongylida). Its LSU rRNA sequence was shorter (by 18 bp) than that of the free-living nematode, Caenorhabditis elegans (order Rhabditida), the only other species within the Nematoda for which a complete LSU rRNA sequence has been determined. Interspecific differences in sequence were greater in the 12 D domains compared with the core segments, with the secondary structure being maintained by partial or complete compensatory base pair changes. The magnitude of interspecific sequence difference in each D domain (except for D6 and D12) was similar, suggesting that several domains contain informative genetic markers for phylogenetic studies of the phylum Nematoda at different taxonomic levels. The LSU rRNA may also provide species-specific markers for the identification of some bursate nematodes of veterinary and medical importance. PMID- 12628594 TI - Significance of novel endothelin-B receptor gene polymorphisms in Hirschsprung's disease: predominance of a novel variant (561C/T) in patients with co-existing Down's syndrome. AB - Several genes have been implicated in the pathogenesis of Hirschsprung's disease (HSCR). In a previous study performed, five novel (V202M, E480K, IVS10-2A/G, D771N, IVS19-9C/T) mutations and one previously described mutation (P937L) have been identified in the RET proto-oncogene in 20% of the study population. To further investigate the involvement of other genes, mutation analysis of the endothelin-B receptor (EDNRB) gene was performed in 52 unrelated sporadic HSCR patients, including 38 non-syndromic and 14 patients with HSCR and Down's syndrome. Six novel (178G/A, 552C/T, 561C/T, 702C/T, IVS3-6C/T and IVS4 + 3A/G) sequence variants and one previously described (831G/A) polymorphism were identified. Statistically significant differences were achieved for six (178G/A, 552C/T, 561C/T, 702C/T, IVS3-6C/T and 831G/A) of these variants. The T-allele of the 561C/T polymorphism was over represented in the HSCR/Down's syndrome patient group (36% representing 5 of 14) compared to normal controls (6% representing 5 of 84) (p < 0.002, chi(2) with Yates correction = 12.14), suggesting that the 561C/T variant is associated with a low penetrance effect in patients with this complex phenotype. Detection of the 178G/A polymorphism in only non-syndromic HSCR patients, provide further support for an important role of specific sequence variants in the EDNRB gene in the HSCR/Down's syndrome phenotype. PMID- 12628593 TI - Development of a 5' nuclease assay to detect ciprofloxacin resistant isolates of the biowarfare agent Yersinia pestis. AB - The detection of antibiotic resistance in agents of biowarfare is extremely critical in order to start appropriate therapy in a timely manner. We have developed a 5' nuclease assay to detect ciprofloxacin resistance (Cip(r)) in Yersinia pestis. Two groups of fluorogenic probes were developed. The first group included a probe homologous to the wild type Y. pestis gyrA sequence with two corresponding probes that were homologous with two different mis-sense mutations in codon 81 of GyrA. The second group of probes included a wild type probe and two corresponding probes that recognized mis-sense mutations in codon 83 or gyrA. These probes specifically reacted only with the homologous DNA sequences. The 5' nuclease assay was sensitive to 1 pg (approximately 1 colony forming unit) of starting template and could be used on semi-purified DNA. We tested our optimized assay against a group of 38 Cip(r) Y. pestis isolates and demonstrated that it was accurate at determining the gyrA allele encoded by these strains. The 5' nuclease assay performed on the LightCycler proved to be sensitive, rapid and accurate at identification of Cip(r) Y. pestis and thus should be useful in characterization of this organism in the event of a future act of bioterrorism. PMID- 12628595 TI - The Ljubljana classification - its application to grading oral epithelial hyperplasia. AB - INTRODUCTION: The diagnosis, prognosis and treatment of particular pathologic entities of the oral mucosa depend almost exclusively on the histologic changes in the epithelium. The basis for the classification of oral epithelial hyperplastic lesions should be their propensity to progress to invasive cancer. MATERIAL AND METHODS: In a retrospective study, 135 biopsies of oral epithelial hyperplastic lesions from 115 patients were classified according to the Ljubljana classification, which has recently been introduced for the grading of laryngeal hyperplastic epithelial lesions. RESULTS: Seventy nine cases (59%) showed simple, 42 (31%) abnormal and 11 (8%) atypical hyperplasia; carcinoma in situ was found in three cases (2%). During the follow-up, ranging from 3 months to 6 years (median 1 year), two cases of atypical hyperplasia progressed to invasive cancer, whereas none of the cases classified as simple or abnormal hyperplasia showed progression. CONCLUSION: The present study suggests that the Ljubljana classification can be reliably applied for classifying oral epithelial hyperplastic lesions into different risk groups, which is essential for prognosis in order to plan therapy. PMID- 12628596 TI - Cephalometric assessment of cranial abnormalities in patients with acromegaly. AB - OBJECTIVE AND PATIENTS: Patients with acromegaly (12 women, 26 men) and a control group (36 women, 50 men) were chosen for cephalometry to assess the size, shape and positional characteristics of the craniofacial bones and the upper airways. RESULTS: When compared with the controls, patients of both sexes with acromegaly were found to have significant anomalies in the orofacial skeleton: increased facial height, elongated ascending ramus mandibulae and greater basion supramentale distance, a negative difference between maxillary and mandibular protrusions, enlarged lower part of the gonion angle and of the angle of inclination of the maxilla, as well as alterations in the neurocranium: enlargement of sella turcica and of sinus frontalis and protrusion of the supraorbital ridges. As for the soft tissues, patients with acromegaly exhibited an elongated soft palate and a diminished angle between the uvular axis and the palatal plane. A comparison between the cephalometric parameters of patients with active acromegaly and those without active disease revealed no significant differences in either sex. CONCLUSION: Patients with acromegaly exhibited an enlargement of all parts of the neurocranium and orofacial bones except the maxilla. The greatest anomaly was seen in the mandible, with greater enlargement of the ascending ramus than of the body of the mandible. The shape of this bone was also altered. PMID- 12628597 TI - The reliability of "Analyze" software in measuring orbital volume utilizing CT derived data. AB - INTRODUCTION: Craniomaxillofacial surgeons require to estimate the orbital volume in a variety of clinical situations. This paper evaluates a new method based on software analysis of computerized tomography (CT) scan data. MATERIAL AND METHODS: Five dried skulls with prosthetic globes and periorbita were non helically scanned in an Elscint 2400 CT scanner. Images obtained were processed using the "Analyze" software package and results compared to the volume of the intraorbital prosthesis as determined by a volume displacement gravimetric method. RESULTS: Estimates of volume produced by the software varied from the gold standard by 0.06-50.44%, with a mean error of 8.8%. CONCLUSION: Despite the use of a variety of scan protocols it was not possible to obtain results with "Analyze" software which were sufficiently accurate for clinical use. PMID- 12628598 TI - Le Fort I sliding osteotomy--a procedure for stable inferior repositioning of the maxilla. AB - INTRODUCTION: Inferior repositioning of the maxilla for correction of vertical maxillary deficiency has been found to be associated with instability and a tendency to relapse. This can not be completely controlled by using bone grafts as relapse commences before complete integration of the graft. METHODS: Improved stability was sought by using rules of geometry when displacing the maxilla down a sloping plane. Anterior and inferior sliding of the maxilla after a double 'M' shaped Le Fort I osteotomy was performed to preserve bony contact at the medial and lateral maxillary buttresses. The plan of this technique was derived from model surgery when true changes at the Le Fort I level were visualized. PATIENTS: 15 patients with class III-malocclusion and anterior vertical deficiency of the maxilla were surgically treated using this technique. RESULTS: Bony contact at the maxillary buttresses after anterior and inferior displacement was possible in patients with less than 6mm movement in a sagittal direction. Only minor skeletal relapse (mean: 0.4mm) was observed in the follow-up period (mean 14.2 months). CONCLUSION: The procedure should be considered whenever vertical maxilla relapse is of concern after anterocaudal displacement. PMID- 12628599 TI - Pulpal blood flow: effects of corticotomy and midline osteotomy in surgically assisted rapid palatal expansion. AB - INTRODUCTION/PURPOSE: Surgically assisted rapid palatal expansion is used for the management of transverse maxillary deficiency in the early permanent dentition. The success depends on the maintenance of an adequate blood supply to the mobilized segments. The aim of this study was to assess the effects of corticotomy and midline osteotomy on the tooth pulpal blood flow. MATERIAL: Laser Doppler flowmetry was applied as a non-invasive and reliable technique for the assessment of pulpal blood flow in the maxillary centrals, canines and first molars. The blood flow was investigated pre- and postoperatively, on the first, third, and seventh postoperative days bilaterally in 13 cases. RESULTS: The results of this study indicate that ischaemia of the pulp can occur following osteotomy at the Le Fort I level. CONCLUSION: Corticotomy 5mm above the dental apices and separating the midpalatal suture did not have any serious effect on pulpal blood flow in this study. PMID- 12628600 TI - Correlation between the arthroscopic diagnosis of synovitis and microvessel density in synovial tissues in patients with internal derangement of the temporomandibular joint. AB - OBJECTIVE: To elucidate the correlation between the arthroscopic diagnosis of synovitis and microvessel density in synovial tissues in patients with internal derangement of the temporomandibular joint (TMJ). STUDY DESIGN: Forty-three joints in 41 patients with internal derangement were examined and biopsies taken. Microvessel density was evaluated using the immunohistochemical method for CD 34 antibody. Arthroscopically diagnosed synovitis was evaluated according to Murakami's criteria. RESULTS: In patients with internal derangement, arthroscopically diagnosed synovitis scores averaged 5.2+/-2.0, according to Murakami et al. (1991). Small to large blood vessels were observed clearly with CD 34 stain. The mean microvessel density was 22.7+/-15.6 per two high power fields (magnification x200). Synovitis scores correlated significantly with microvessel density (p=0.002, r=0.43). CONCLUSION: Synovitis evaluated using Murakami's scores correlated well with the number of blood vessels in synovial tissues in patients with internal derangement of the TMJ. This demonstrates that synovitis is linked to inflammation-related blood vessel density of the synovial tissues. PMID- 12628601 TI - The use of polylactic acid/polyglycolic acid copolymer and gelatin sponge complex containing human recombinant bone morphogenetic protein-2 following condylectomy in rabbits. AB - PURPOSE: To examine the results of a polylactic acid/polyglycolic acid copolymer and gelatin sponge complex (PGS) with or without recombinant human bone morphogenetic protein-2 (rhBMP-2) used to treat condylar defects in rabbits. MATERIAL AND METHODS: Adult male Japanese white rabbits (n=60; 3kg; 12-16 weeks old) were divided into three groups of 20 each. All rabbits underwent condylectomy. In the two implanted groups, PGS with or without 5 microg of rhBMP 2 was implanted to the condylar defect without fixation. No material was implanted into the control group. Animals were sacrificed at 2, 4, 8, 12 and 24 weeks postoperatively, and the temporomandibular joints (TMJs) were examined histologically. RESULTS: Four weeks after implantation, growth of bone and cartilage-like tissue was observed in all rabbits that received PGS implants (with and without rhBMP-2). A cartilage-like layer was derived from the bone marrow at the operated surface. There was no growth of bone tissue in the control rabbits, but they also had a cartilage-like layer directly derived from the operated surface. CONCLUSION: This study demonstrated that PGS with or without rhBMP-2 could induce regeneration of new bone and cartilage-like tissue in the TMJ. PMID- 12628602 TI - Replantation of an avulsed zygomatic bone as a freeze-preserved autologous graft: a case report. AB - We describe a very rare case of midface fracture where the zygomatic bone had been completely avulsed. The fragment was preserved in a frozen state for 40 days, then replanted and the graft was taken. The conditions favourable for the taking of the freeze-preserved zygomatic graft were: (1) an appropriate preservation method, (2) conservation of the periosteum on the zygomatic bone, (3) diversity of the haemodynamics of the zygomatic membranous bone, (4) immobilization of the zygoma by rigid fixation, and (5) a rich blood supply at the recipient site. PMID- 12628603 TI - Evaluation of stress patterns generated by reduction forceps within a photoelastic mandibular model. AB - INTRODUCTION: Little attention has been paid to the mechanical effects of fracture reduction forceps. AIM: This study aims to evaluate the stress patterns within the fractured mandible generated by reduction forceps. MATERIAL AND METHODS: Thirty-six mandibular models were fabricated using a photoelastic resin. Each of the three sets of mandibular models was osteotomized according to one of three different fracture types. After reducing the cut segments, reduction forceps were placed into different engagement holes to compress the segments. Photoelastic stress analysis was used to visualize the stress patterns within the fractured mandibular models as generated by the reduction forceps. RESULTS: In the case of symphyseal or parasymphyseal fractures, an optimum distribution of stresses over the fracture site was achieved when placing the reduction forceps more than 12mm away from either side of the fracture line, between the midway level of the mandibular height (bisecting the mandible) and 5mm below this level. In the case of body fractures, optimum stress distribution was achieved when the reduction forceps were placed more than 16mm from the fracture line at the midway level. CONCLUSION: Correct use of the reduction forceps helps to provide a precise three-dimensional reduction for mandibular fractures. PMID- 12628604 TI - A complex-valued nonlinear neural adaptive filter with a gradient adaptive amplitude of the activation function. AB - A complex-valued nonlinear gradient descent (CNGD) learning algorithm for a simple finite impulse response (FIR) nonlinear neural adaptive filter with an adaptive amplitude of the complex activation function is proposed. This way the amplitude of the complex-valued analytic nonlinear activation function of a neuron in the learning algorithm is made gradient adaptive to give the complex valued adaptive amplitude nonlinear gradient descent (CAANGD). Such an algorithm is beneficial when dealing with signals that have rich dynamical behavior. Simulations on the prediction of complex-valued coloured and nonlinear input signals show the gradient adaptive amplitude, CAANGD, outperforming the standard CNGD algorithm. PMID- 12628605 TI - Self-organizing continuous attractor networks and motor function. AB - Motor skill learning may involve training a neural system to automatically perform sequences of movements, with the training signals provided by a different system, used mainly during training to perform the movements, that operates under visual sensory guidance. We use a dynamical systems perspective to show how complex motor sequences could be learned by the automatic system. The network uses a continuous attractor network architecture to perform path integration on an efference copy of the motor signal to keep track of the current state, and selection of which motor cells to activate by a movement selector input where the selection depends on the current state being represented in the continuous attractor network. After training, the correct motor sequence may be selected automatically by a single movement selection signal. A feature of the model presented is the use of 'trace' learning rules which incorporate a form of temporal average of recent cell activity. This form of temporal learning underlies the ability of the networks to learn temporal sequences of behaviour. We show that the continuous attractor network models developed here are able to demonstrate the key features of motor function. That is, (i) the movement can occur at arbitrary speeds; (ii) the movement can occur with arbitrary force; (iii) the agent spends the same relative proportions of its time in each part of the motor sequence; (iv) the agent applies the same relative force in each part of the motor sequence; and (v) the actions always occur in the same sequence. PMID- 12628606 TI - Zonal function network frames on the sphere. AB - We introduce a class of zonal function network frames suitable for analyzing data collected at scattered sites on the surface of the unit sphere of a Euclidean space. Our frames consist of zonal function networks and are well localized. The frames belonging to higher and higher scale wavelet spaces have more and more vanishing polynomial moments. The main technique is applicable in the general setting of separable Hilbert spaces, in which context, we study the construction of new frames by perturbing an orthonormal basis. PMID- 12628607 TI - On motion detection through a multi-layer neural network architecture. AB - A neural network model called lateral interaction in accumulative computation for detection of non-rigid objects from motion of any of their parts in indefinite sequences of images is presented. Some biological evidences inspire the model. After introducing the model, the complete multi-layer neural architecture is offered in this paper. The architecture consists of four layers that perform segmentation by gray level bands, accumulative charge computation, charge redistribution by gray level bands and moving object fusion. The lateral interaction in accumulative computation associated learning algorithm is also introduced. Some examples that explain the usefulness of the system we propose are shown at the end of this article. PMID- 12628608 TI - Dual extended Kalman filtering in recurrent neural networks(1). AB - In the classical deterministic Elman model, the estimation of parameters must be very accurate. Otherwise, the system performance is very poor. To improve the system performance, we can use a Kalman filtering algorithm to guide the operation of a trained recurrent neural network (RNN). In this case, during training, we need to estimate the state of hidden layer, as well as the weights of the RNN. This paper discusses how to use the dual extended Kalman filtering (DEKF) for this dual estimation and how to use our proposing DEKF for removing some unimportant weights from a trained RNN. In our approach, one Kalman algorithm is used for estimating the state of the hidden layer, and one recursive least square (RLS) algorithm is used for estimating the weights. After training, we use the error covariance matrix of the RLS algorithm to remove unimportant weights. Simulation showed that our approach is an effective joint-learning pruning method for RNNs under the online operation. PMID- 12628609 TI - Kalman filters improve LSTM network performance in problems unsolvable by traditional recurrent nets. AB - The long short-term memory (LSTM) network trained by gradient descent solves difficult problems which traditional recurrent neural networks in general cannot. We have recently observed that the decoupled extended Kalman filter training algorithm allows for even better performance, reducing significantly the number of training steps when compared to the original gradient descent training algorithm. In this paper we present a set of experiments which are unsolvable by classical recurrent networks but which are solved elegantly and robustly and quickly by LSTM combined with Kalman filters. PMID- 12628610 TI - Intelligent optimal control with dynamic neural networks. AB - The application of neural networks technology to dynamic system control has been constrained by the non-dynamic nature of popular network architectures. Many of difficulties are-large network sizes (i.e. curse of dimensionality), long training times, etc. These problems can be overcome with dynamic neural networks (DNN). In this study, intelligent optimal control problem is considered as a nonlinear optimization with dynamic equality constraints, and DNN as a control trajectory priming system. The resulting algorithm operates as an auto-trainer for DNN (a self-learning structure) and generates optimal feed-forward control trajectories in a significantly smaller number of iterations. In this way, optimal control trajectories are encapsulated and generalized by DNN. The time varying optimal feedback gains are also generated along the trajectory as byproducts. Speeding up trajectory calculations opens up avenues for real-time intelligent optimal control with virtual global feedback. We used direct-descent curvature algorithm with some modifications (we called modified-descend controller-MDC algorithm) for the optimal control computations. The algorithm has generated numerically very robust solutions with respect to conjugate points. The adjoint theory has been used in the training of DNN which is considered as a quasi-linear dynamic system. The updating of weights (identification of parameters) are based on Broyden-Fletcher-Goldfarb-Shanno BFGS method. Simulation results are given for an intelligent optimal control system controlling a difficult nonlinear second-order system using fully connected three-neuron DNN. PMID- 12628612 TI - Neural network based tracking control of a flexible macro-micro manipulator system. AB - In this paper, we address the problem of stable tracking control of a flexible macro-micro manipulator (M(3)) system. A two-layer neural network is utilized to approximate the nonlinear robot dynamic behavior of the M(3) system, and the controllers for the macro and micro arms are developed without any need for prior knowledge of the dynamic model of the controlled M(3) system. A learning algorithm for the neural network using Lyapunov stability theory is derived. It is shown that both the tracking error and the weight-tuning error are uniformly ultimately bounded under this new control scheme. Simulation results are presented and compared to those obtained using a PD controller. PMID- 12628611 TI - Clustering ensembles of neural network models. AB - We show that large ensembles of (neural network) models, obtained e.g. in bootstrapping or sampling from (Bayesian) probability distributions, can be effectively summarized by a relatively small number of representative models. In some cases this summary may even yield better function estimates. We present a method to find representative models through clustering based on the models' outputs on a data set. We apply the method on an ensemble of neural network models obtained from bootstrapping on the Boston housing data, and use the results to discuss bootstrapping in terms of bias and variance. A parallel application is the prediction of newspaper sales, where we learn a series of parallel tasks. The results indicate that it is not necessary to store all samples in the ensembles: a small number of representative models generally matches, or even surpasses, the performance of the full ensemble. The clustered representation of the ensemble obtained thus is much better suitable for qualitative analysis, and will be shown to yield new insights into the data. PMID- 12628616 TI - Alcoholism treatment outcome studies, 1970-1998. An expanded look at the nature of the research. AB - We reviewed 701 alcohol treatment outcome studies reported between 1970 and 1998. Examining the nature of the research allows us to characterize this field of study, analyze trends over time, and improve upon this work in the future. For multiple-group studies, the majority of both investigators and participants were male, and participant characteristics were generally inconsistently surveyed and reported. Between the 1970s and the 1990s, research on pharmacological treatments and research conducted in outpatient settings became more common. Additionally, the types of outcome measures assessed varied more widely and were more likely to be continuous and quantifiable. PMID- 12628615 TI - Role of self-help processes in achieving abstinence among dually diagnosed persons. AB - The effectiveness of participation in dual-focus groups (i.e., focusing on both mental health and substance use) has not been studied empirically. The study examined whether three hypothesized active ingredients of self-help (helper therapy, reciprocal-learning, and emotional-support processes) are associated with drug/alcohol abstinence outcomes for members of a 12-step dual-focus fellowship, Double Trouble in Recovery (DTR). The study was able to control for member attitudes and behaviors at baseline, which might be related to both self help processes and outcomes, i.e., extent of participation in DTR and traditional 12-step groups, prior drug/alcohol use, severity of psychiatric symptoms, motivation for change, stressful life events, perceived coping, self-efficacy for recovery, and social support. Members of 24 DTR groups in New York City were recruited, interviewed, and reinterviewed after 1 year. Drug/alcohol abstinence in the past year increased from 54% at baseline to 72% at follow-up. Helper therapy and reciprocal-learning activities were associated with better abstinence outcomes, independent of other attitudes and behaviors of the members. However, emotional support was not related to outcome. We conclude that specific elements of self-help participation contribute substantially to progress in recovery for members of dual-focus groups; facilitating such self-help processes should be encouraged by clinicians and senior fellowship members. PMID- 12628617 TI - A pilot trial of piracetam and ginkgo biloba for the treatment of cocaine dependence. AB - BACKGROUND: Chronic cocaine use is associated with cognitive deficits that may reduce the effectiveness of psychosocial treatment and promote relapse in newly abstinent cocaine-dependent patients. Nootropic agents, such as piracetam and ginkgo biloba, may improve cognitive function and reduce the incidence of relapse in these patients. METHODS: This was a 10-week, double-blind, placebo-controlled pilot trial involving 44 cocaine-dependent subjects. Subjects received either piracetam (4.8 g/day), ginkgo biloba (120 mg/day), or placebo. Subjects were required to attain abstinence from cocaine during a 2-week baseline phase demonstrated by providing at least one benzoylecgonine (BE)-negative urine toxicology screen. Outcome measures included treatment retention, urine toxicology screens, Clinical Global Impression (CGI) scores, and results from the Addiction Severity Index (ASI). RESULTS: Ginkgo biloba was not superior to placebo in any outcome measure. Piracetam was associated with more cocaine use and lower CGI scores compared to placebo. CONCLUSIONS: Neither piracetam nor ginkgo biloba appears to be a promising medication for the treatment of cocaine dependence. PMID- 12628618 TI - Alcohol relapse as a function of relapse definition in a clinical sample of adolescents. AB - Relapse is an important clinical and research problem in the addictions, but one that has no consensus on an operational definition. How relapse is measured could make substantial differences in clinical practice and in conclusions drawn from empirical studies. The aim of this study was to compare and contrast four different operational definitions of alcohol relapse in a clinical sample of adolescents. The participants were 75 males and females ages 14-18 who presented for outpatient treatment of alcohol use disorder (AUD). They completed an extensive in-person initial assessment, as well as 12 subsequent monthly telephone interviews that concerned their daily alcohol and other drug use and related events. The four alcohol relapse definitions evaluated included: at least 1 day of drinking any amount after at least 4 consecutive days of abstinence; at least 1 heavy (five standard drinks for boys, four for girls) drinking day after 4 abstinent days; at least 1 day of drinking any amount with associated problems after 4 abstinent days; and at least 1 heavy drinking day with associated problems following 4 abstinent days. Relapse events were identified during the first 6 months following the initial interview. The results showed that the time period until relapse varied across the four definitions of relapse ranging from 26 to 90 days. Additional analyses investigated the utility of each of the definitions in predicting functioning during months 7-12 using multiple regression. The results of these analyses showed that the two "heavy drinking" definitions contributed significantly independent variance to predicting average number of drinking days/month and drinks/drinking day and that the two "any drinking" definitions significantly predicted the presence of a current AUD diagnosis. The results suggest that relapse definitions make a difference in estimates of rates and times to first relapse. Furthermore, the occurrence of at least one relapse is predictive of later functioning in a clinical sample of adolescents, which is of practical and theoretical importance. Suggestions for the extensions of this investigation in future research are provided. PMID- 12628619 TI - Onset of major depression during treatment for nicotine dependence. AB - We monitored the emergence of major depression (MDD) during treatment for nicotine dependence among 224 smokers. MDD was assessed on three occasions during the course of treatment with the mood disorders portion of the Structured Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorders (SCID), fourth edition (DSM-IV). Out of 224 participants, 20% had suffered a past episode of MDD, 18% of males and 22% of females. Four percent (n=10) experienced onset of MDD during the course of the study, four males and six females. Only 2 of the 10 cases managed to achieve abstinence at end of treatment. Those who reported large increases in depression symptoms between baseline and end of treatment (Week 10) were less likely to be abstinent at 26-week follow-up. The evidence indicates that those who treat nicotine dependence must be prepared to monitor and respond to the emergence of depression associated with treatment. PMID- 12628620 TI - Differences in evaluations of a tobacco awareness and cessation program by adolescents in four stages of change. AB - This study determined whether adolescents in four stages of change provided significantly different ratings on evaluations of a tobacco awareness and cessation program and the program facilitators. A systematic sample of 639 adolescents in four stages of change (precontemplation, contemplation, preparation, and action) was used for analysis. Analysis of variance revealed significant differences in ratings on 'Cessation Skills,' 'Situation Management,' 'Overall Facilitator,' and 'Facilitator Likeability' evaluations between the groups. Follow-up analysis revealed that adolescent precontemplators provided significantly lower ratings than those in preparation and action, and adolescents in action provided significantly higher ratings than precontemplators and contemplators. This study suggests a need to assess the stage of the adolescent at the beginning of the program in order to provide the facilitator the opportunity to engage and motivate adolescents who are in the precontemplation and contemplation stage of change. This study also provides important information for facilitator training, program planning, and program delivery. PMID- 12628621 TI - Substance use in the general hospital. AB - INTRODUCTION: People misusing substances are overrepresented in health settings. Substance misuse can also be an underlying factor complicating medical diagnosis and management. AIMS: (i) To establish the prevalence of substance misuse in the general hospital inpatient population; and (ii) to examine the relationship between medical diagnosis and substance misuse problem as identified by screening tools. METHOD: This study adopted a three-step screening approach including a prospective questionnaire survey, interview, and case note review. Subjects included all adult patients admitted to a London teaching hospital over a 1-week period. RESULTS: Seventy percent completed the questionnaire. Twenty-three percent was currently smoking, 14% was rated positive for alcohol misuse, and 12% positive for drug misuse. Only 65% of patients was screened for smoking, 54% for alcohol, and 9% for other substances. CONCLUSIONS: People misusing one substance were more likely to be misusing others, hence the importance of screening all patients for all substances. Doctors were screening a small percentage of patients and they identified and intervened with only the severe ones. PMID- 12628622 TI - A prospective evaluation of the relationships between smoking dosage and body mass index in an adolescent, biracial cohort. AB - Although there is clearly an inverse relationship between smoking and body weight, recent studies suggest that weight attenuation via smoking is slow and may take decades to accrue. This investigation prospectively evaluated the relationships between smoking dosage (or lack thereof) and relative weight change in 1697 adolescents followed over 4 years. A 4 (smoking groups: 0, 1, 2, or 3 or more years of smoking exposure)x2 (ethnicity: Caucasian or African American)x2 (gender: male or female) analysis of variance (ANOVA) was performed to assess weight gain attenuation associated with increasing exposure to smoking. The overall results revealed a significant three-way interaction between smoking dosage, gender, and ethnicity. Specifically, smoking initiation was associated with an increase in body mass index (BMI) for 2 years after initiation. For those youth smoking 3 or more years, body weights were almost identical compared to never-smokers. No significant reductions in body weight were observed in any gender or ethnic group for up to 3 years after smoking initiation. It is concluded that smoking initiation is not associated with adolescent body weight change for at least a 3-year period. PMID- 12628623 TI - Smoking differences among African American, Hispanic, and White middle school students in an urban setting. AB - INTRODUCTION: Cross-sectional studies have repeatedly observed that African American adolescents are less likely to smoke when compared to White and Hispanic adolescents. Although much is known among high school samples, few scientifically based studies have reported these race and ethnic differences in cigarette smoking among younger samples. METHODS: This study employed a secondary analysis of data from a 3-year middle school violence prevention project. The study design was a 3-year serial cross-sectional survey, out of which a cohort of students from sixth to eighth grade was formed. Smoking measurers were taken yearly. RESULTS: 8865 students responded in 1994, 9115 in 1995, and 9364 in 1996; 1589 students are in the 3-year cohort. Smoking prevalence rates from both measurement periods confirm the disparity between African Americans, Whites, and Hispanic youth. Although weak in sixth grade, by eighth grade, White and Hispanic students are smoking at two to four times the rate of their African American classmates. CONCLUSIONS: Clearly, more etiological research needs to be conducted to understand the social, cultural, and intrapersonal forces that operate to inhibit the onset of smoking in African American youth and promote the onset of smoking in White and Hispanic youth. PMID- 12628624 TI - Contextual factors and other correlates of sexual risk of HIV among African American crack-abusing women. AB - This study examined differences in contextual factors, substance use, sexual risk behaviors, and comorbid histories between African-American, out-of-treatment, crack-abusing women who had either a single sexual partner or multiple partners. Bivariate analysis indicated that women with multiple partners were more likely than women with a single partner to be homeless, financially dependent, and to have histories of sexual, physical, and emotional abuse. Women with multiple partners reported higher levels of depression, anxiety, and more symptoms of posttraumatic stress disorder (PTSD). In multiple logistic regression analysis, being unemployed, difficult childhood, and number of days of crack use in the previous 30 days, longer crack runs, and more frequent unprotected fellatio were associated with increased odds of having multiple sexual partners. Being married or living as married was associated with decreased odds of having multiple sexual partners. The importance of assessing contextual and historical factors and implications for future research is discussed. PMID- 12628625 TI - A model of the effects of node-link mapping on drug abuse counseling. AB - This paper examines the effectiveness of a promising cognitive enhancement technique called node-link mapping. Maps provide a concrete, visual, and computationally efficient means for exploring personal issues, and have received substantial empirical support. We introduce two models to provide a framework for understanding how node-link mapping improves substance abuse counseling. The TCU Process Model reviews the general process of substance abuse treatment. The second model reflects an effective treatment session. Implications of the models and node-link mapping findings are discussed, as well as future research directions. PMID- 12628626 TI - Reinforcement expectancies for studying predict drinking problems among college students: approaching problem drinking from an expectancies choice perspective. AB - Molar behavioral choice theories point to the important role reinforcement for alternative behaviors may play in reducing problem drinking [Psychol. Addict. Behav. 9 (1995) 223.]. A similar molar choice perspective may apply to a cognitive mediational level of analysis: the molecular link between alcohol expectancies and drinking behavior depends on expectancies for alternative behaviors. In this study, 82 college students completed the Alcohol Effects Questionnaire (AEFQ), the Rutgers Alcohol Problem Index (RAPI), a self-report measure of drinking quantity and frequency, and a five-item studying expectancies questionnaire devised for the current study. Analyses indicated that, among students who hold high alcohol expectancies, those who reported high studying expectancies drink less and develop significantly fewer drinking problems than those who hold low studying expectancies. Future research may address the possibility of reducing drinking problems among college students by enhancing studying expectancies. PMID- 12628627 TI - The influence of cognitive coping and mood on smokers' self-efficacy and temptation. AB - Previous descriptive and treatment studies imply that the association between depressed affect and cigarette smoking may be strongest among those with limited cognitive coping skills. This study, therefore, experimentally examined whether the combination of poor mood management skills and negative affect results in reduced self-efficacy and increased temptation to smoke. Current smokers were randomly assigned to an elated-mood or a depressed-mood induction condition. State mood, temptation, and self-efficacy were measured before and after the induction. Contrary to prediction, mood induction condition did not interact with cognitive coping skill to predict change in self-efficacy or temptation. However, there was a significant interaction of (measured) state happiness and "positive" (functional according to expert cognitive therapists) responses on the Ways of Responding (WOR) test of coping skills in predicting temptation: those with low levels of positive coping responses and low positive affect after the induction were especially tempted to smoke. This latter finding suggests that smokers with a history of depression may respond well to interventions aimed at increasing positive affect and augmenting positive cognitive coping skills. PMID- 12628629 TI - A mediated moderation model of cigarette use among Mexican American youth. AB - The current study tested a model examining both the direct and mediated effects of Ethnic Cultural Norms (ECN) on cigarette use in a sample of Mexican American youth (ages 11-14; N=921). Contextual risk factors (peer smoking and family smoking) were included as potential moderators of this mediational relationship. A product of coefficient (alpha beta) method to test the significance of the mediated effect [Evaluation Review 17 (1993) 144.] was adapted to assess the mediation of interaction effects. Results suggested that Tobacco Avoidance Self Efficacy mediated the protective influence of ECN on cigarette use. However, as peer smoking increased, the influence of ECN on cigarette use diminished, though it remained a significant, protective influence on cigarette use. Results are discussed in terms of the potential synergy between ECN and social influence training in tobacco preventive intervention development among youth of Latin descent. PMID- 12628628 TI - Patterns of depressive symptomatology in women smokers, ex-smokers, and never smokers. AB - To investigate the possibility that specific symptom patterns contribute differentially to excess depression among smokers, we analyzed Center for Epidemiological Studies-Depression (CES-D) and subscale scores in 931 women current smokers (CS), ex-smokers (ES), and never-smokers (NS). After adjusting for differences in age, education, and marital status, significant group differences were found for the CES-D overall and for Depressed Affect, Anhedonia, and Somatic Features, but not Interpersonal Distress. Both CS and ES scored significantly higher than NS on most measures, whereas differences between CS and ES failed to reach significance. CS were significantly more likely than NS to be taking antidepressants. Alcohol intake also showed significant group differences, with CS>ES>NS, suggesting that the link between current smoking and alcohol intake is mediated by factors other than depression, since CS and ES were comparably depressed. We conclude that studying specific symptom patterns may elucidate the smoking-depression link. Our observation that ES and CS exhibit similar levels of depressive symptomatology further suggests that interventions for depressed smokers need to take into account the likely persistence of depression following cessation. PMID- 12628631 TI - Getting by with a little help from your friends: the impact of peer networks on criminality in a cohort of treatment-seeking drug users. AB - This study investigates the links between social networks of drug users and criminal activity. Opiate misusers (n=128) receiving in-patient treatment were interviewed about substance use, social networks, and crime in the month before treatment. Almost 60% of participants reported an average of more than 70 crimes each. Less than one-fifth of the subjects spent no time with other users, while just over half spent either "quite a lot" or "a lot" of time with drug users. Time with users increased the risk of crime. Spending no time with users provided a protective effect. Time with criminally involved drug users was associated with greater levels of crime. Social networks represent an important marker for integration in criminal networks in treatment-seeking drug users and a long-term barrier to rehabilitation. PMID- 12628630 TI - Prevalence of substance use among offspring of opioid addicts. AB - AIMS: This study assessed the prevalence of substance use in sons and daughters of opioids addicts in Iran. MEASUREMENTS: A confidential semistructured interview and DSM-IV criteria were used. Data were gathered from 500 subjects (225 men and 275 women). These subjects were selected randomly from sons and daughters of 500 opioids addicts (out of 1500 opioids addicts) who sought treatment in 2000. RESULTS: Of these subjects, 33.8% were in the age range of 20-29, 33.1% in 30-39, 28% were >40, and 7.1% were <20 years; of them, 81.6% were married and 18.4% were single. The majority (36.2%) used cigarettes, 19% used opium, 18.2% used alcohol, 2.6% used hashish, and 1.6% used marijuana once or more at some time in their lives. Nobody used cocaine or psychedelics. Of the current substance dependents, using DMS-IV criteria, 46.7% of men and 5.5% of women were nicotine dependent, 12.9% of men and 1.1% of women were opium dependent, and 0.9% of men and 0.7% of women were heroin dependent. (Some used or were using more than one substance.) Enjoyment (56.8%) was the first and modeling (49.6%) was the second most common reason reported for substance use. CONCLUSIONS: Substance use was significantly higher in men, in single, and also in illiterate subjects. There was a nonsignificant relation between age group and substance use. Cultural attitudes toward drug use quite likely affect the type and amounts of use. These findings can be considered when planning preventive programs. PMID- 12628632 TI - Neuropsychological correlates of opioid dependence and withdrawal. AB - Severity of opioid dependence, and performance on two successive runs of the Wisconsin Card Sorting Test (WCST), were assessed in 39 right-handed male and female methadone patients who had been randomly assigned to either a recently dosed (n=21) or 24 hr abstinent (n=18) condition. Results indicated that severity of opioid dependence was positively correlated with perseverative responses and errors on the second run of the WCST, p<.05. Further, controlling for the effect of dependence severity, patients in early methadone withdrawal made selectively more perseverative responses and errors than did recently dosed patients, p<.05, with no difference on nonperseverative errors. Findings were consistent with the hypothesis that opioid dependence, like alcoholism and cocaine addiction, is associated with disruption of executive cognitive functions mediated by the prefrontal cortex. PMID- 12628634 TI - Cross-cultural validity of the Yale-Brown Obsessive Compulsive Scale. AB - Some psychometric properties of an adaptation of the Yale-Brown Obsessive Compulsive Scale for use in the Netherlands (Y-BOCS-NL) were examined in 65 psychiatric inpatients. The factorial invariance of two-dimensional systems were determined, namely Severity and Disturbance versus Obsessions and Compulsions, with the latter performing substantially better than the former in different respects. All further analyses were therefore focused on the Obsessions and Compulsions prototype. Inter-rater agreement was established in four inpatients by 39 raters at different institutions and found to be excellent. Internal consistency was also excellent. Concurrent and divergent validity were examined and established by correlating the Y-BOCS-NL with the Padua Inventory, the Burns Perfectionism Scale and state and/or trait measures of, among others, hostility, anxiety, depression, irrational fears and assertiveness. The Obsessions and Compulsions dimensions were lowly intercorrelated and showed evidence of clearly different patterns of correlations with external measures (divergent validity). PMID- 12628633 TI - Treatment of childhood anxiety disorders: a preliminary comparison between cognitive-behavioral group therapy and a psychological placebo intervention. AB - The present study examined the efficacy of group cognitive-behavioral therapy (CBT) in the treatment of childhood anxiety disorders. Thirty high-anxious children (aged 9-12 years) were assigned to either (a) group CBT (n = 10), (b) a psychological placebo intervention (i.e., emotional disclosure [ED]; n = 10), or (c) a no-treatment control condition (n = 10). Therapy outcome measures (i.e., children's self-report of anxiety disorders symptoms, depression, and trait anxiety) were obtained three months before treatment, at pretreatment, and at posttreatment. Results showed that levels of psychopathological symptoms remained relatively stable during the three months preceding treatment. Most importantly, pretreatment-posttreatment comparisons indicated that CBT was superior to ED and the no-treatment control condition. That is, only in the CBT condition significant reductions of anxiety disorders symptoms, trait anxiety, and depression were observed. These findings can be taken as further evidence for the efficacy of CBT in the treatment of childhood anxiety disorders. PMID- 12628635 TI - Instructed neutralization, spontaneous neutralization and prevented neutralization after an obsession-like thought. AB - Building on two earlier experiments (Behav. Res. Ther. 34 (1996) 889; 39 (2001) 1439) the present study investigated the effects of neutralizing the consequences of an obsession-like thought in healthy participants. Just like in the earlier studies, writing out and thinking over such a thought generated anxiety. After this provocation, 40 of the 120 participants were instructed to neutralize the effects of the thought for 2 min, 40 participants did not receive a particular instruction, and the remaining 40 participants were instructed to do mental arithmetic aloud so as to prevent "spontaneous" attempts at neutralizing the thought. The no instruction group reported that they neutralized (spontaneously) to the same degree as the group that was instructed to neutralize. Within 2 min, anxiety decreased to near base line levels and there were no differences between the three conditions. When the groups were asked to bring the obsession-like thought back to consciousness again, anxiety increased slightly. Yet, contrary to expectation, this increase in anxiety did not discriminate the "neutralization prevention" group from the other two groups. Limitations of the paradigm as a model for clinical obsessions are discussed. PMID- 12628636 TI - Covariation bias in the affect-modulated startle paradigm. AB - A covariation bias (CB) reflects the overestimation of an objectively random covariation between fear-relevant picture stimuli (e.g., spiders) and aversive consequences (e.g., electric shocks). The affect-modulated startle (AMS) paradigm is similar to it because aversive reflex probes are delivered in the context of unpleasant, neutral or pleasant picture stimuli. The present study explored if a covariation bias is present in the affect-modulated startle paradigm. There was a covariation bias for unpleasant and pleasant compared to neutral pictures probably due to differences in arousal. Startle reflexes were modulated by picture valence. No correlation between a covariation bias index and the startle response modulation was found. It is concluded that both phenomena are mediated by different underlying mechanisms. PMID- 12628637 TI - Does graded activity increase activity? A case study of chronic fatigue syndrome. AB - The reliance on self-report outcome measures in clinical trials of graded activity-oriented cognitive-behavior therapy in chronic fatigue syndrome (CFS) makes it difficult to draw definitive conclusions about actual behavioral change. The participant in this case study was a 52-year-old married male with CFS who was working full-time. Outcome measures included a step counter to objectively measure physical activity as well as a daily diary measure of exercise activity and in vivo ratings of perceived energy, fatigue, and affect. The following psychometric instruments were also used: the CFS Symptom Inventory, the SF-36, the Beck Depression Inventory, and the Beck Anxiety Inventory. The 26-session graded activity intervention involved gradual increases in physical activity. From baseline to treatment termination, the patient's self-reported increase in walk time from 0 to 155 min a week contrasted with a surprising 10.6% decrease in mean weekly step counts. The final follow-up assessment revealed a "much improved" global rating, substantial increases in patient-recorded walk time and weight lifting intensity, yet a relatively modest increment in weekly step counts. It appeared that improvement was associated with mood-enhancing, stress reducing activities that were substituted for stress-exacerbating activities. PMID- 12628638 TI - A multiple-baseline evaluation of the treatment of food phobia in a young boy. AB - A multiple-baseline design was used to evaluate the effectiveness of a behavioral treatment program using modeling, graduated exposure, and contingency management to treat food phobia in a 4-year-old boy. In addition, a treatment component involving time-out and re-introduction of the initial request to consume the target food was added to reduce vomiting behavior that developed during the course of treatment. The volume and range of foods consumed by the participant increased, and observer-rated anxiety and vomiting decreased over the course of treatment. The results of this controlled evaluation suggest that this treatment program was responsible for the observed changes, which were maintained at 6 month follow-up. PMID- 12628642 TI - Synthesis and biological evaluation of substituted 2-sulfonyl-phenyl-3-phenyl indoles: a new series of selective COX-2 inhibitors. AB - A new series of substituted 2-sulfonyphenyl-3-phenyl-indole derivatives were synthesized and evaluated for their ability to inhibit COX-2 and COX-1enzymes. Most of the compounds synthesized were found to be highly potent and selective inhibitors of COX-2. This work led to the discovery of 2-aminosulfonylphenyl-3 phenyl-indole 5a which possesses higher activity and selectivity for COX-2 than Celecoxib both in vitro and in vivo. PMID- 12628643 TI - Synthesis and biological evaluation of five-membered heterocycles fused to cyclopenta[c]thiophene as new antitumor agents. AB - A series of 10 derivatives 2-6 issued from the fusion of various five-membered heterocycles to cyclopenta[c]thiophene were evaluated for potential anticancer activity in the NCI's in vitro human disease-oriented tumor cell line screening panel that consisted of 60 human tumor cell lines arranged in nine subpanels, representing diverse histologies. Among these tested compounds, four were found to be cytotoxic allowing us to point out some structure-activity relationships. The oxazolidinone derivatives 2a-c displayed further in vivo antitumor activity in the hollow fiber assay and standard xenograft testing developed at the NCI. PMID- 12628644 TI - Synthetic studies on himbacine, a potent antagonist of the muscarinic M2 subtype receptor. Part 2: synthesis and muscarinic M2 subtype antagonistic activity of the novel himbacine congeners modified at the C-3 position of lactone moiety. AB - With an aim to disclose the convergency and flexibility of our previously explored synthetic route to natural himbacine 1, and moreover, to clarify some novel aspects of the structure-activity relationships of 1, we prepared various structural types of novel himbacine congeners, 3-demethylhimbacine (3 norhimbacine) 2 and 4-epi-3-demethylhimbacine (4-epi-3-norhimbacine) 4-epi-2 and their enantiomers (ent-2 and ent-4-epi-2), 11-methylhimbacine 3, and 3 epihimbacine 4 in optically pure forms by employing our methodology. All of the synthesized congeners correspond to the compounds modified at the C-3 position of gamma-lactone moiety involved in 1. Among these congeners, 3-demethylhimbacine (3 norhimbacine) 2 was found to exhibit more potent muscarinic M(2) receptor binding affinity than natural 1. PMID- 12628645 TI - Pharmaco-toxicological study of diterpenoids. AB - Azorella compacta, Azorella yareta and Laretia acaulis (Apiaceae) are native species from the high Andes Mountains, northeastern Chile, and they have being traditionally used to treat asthma, colds and bronchitis, illnesses with inflammation and pain as the main symptoms. Interestingly, there are no scientific reports available on their benefits or toxicity. This study was carried out with the purpose of validating the medicinal use of these species and to discover anti-inflammatory and analgesic new molecules. As a working hypothesis, we have proposed that these medicinal species contain bioactive compounds with anti-inflammatory and analgesic effects. In this context, azorellanol, 13-hydroxy-7-oxoazorellane and 7-deacetylazorellanol, three diterpenoids previously isolated only from these plants, were subjected to farmaco-toxicological evaluation. Their topical anti-inflammatory and analgesic activities along with acute toxicities or innocuosness were also investigated. Our results indicate the absence of toxic and side effects in mice. All compounds presented dose-related inhibition of pain. 13-hydroxy-7-oxoazorellane was the most potent analgesic but it was less effective than sodium naproxen, the reference drug. Azorellanol exhibited the highest topical anti-inflammatory potency on AA (arachidonic acid) and TPA (12-deoxyphorbol 13-tetradecanoate) induced oedema, and it effect was similar to the reference drugs (nimesulide and indomethacin). Probably, its mechanism of action could be explained through the inhibition to cyclo-oxygenase activity. Our results corroborate the anti inflammatory and analgesic effects of these species, and it justifies their use in folk medicine. PMID- 12628646 TI - Protection against Leishmania major infection by oligomannose-coated liposomes. AB - Liposomes coated with neoglycolipids constructed with mannopentaose and dipalmitoylphosphatidylethanolamine (Man5-DPPE) have been shown to induce cellular immunity against antigens encapsulated in the liposomes. To assess whether these neoglycolipid-coated liposomes can elicit protective immune response against challenge infection, effects of immunization with soluble leishmanial antigens encapsulated in the liposomes were evaluated using Leishmania major infection in susceptible BALB/c mice. Intraperitoneal immunization of mice with leishmanial antigens in the Man5-DPPE-coated liposomes significantly suppressed footpad swelling in comparison to the control, non immunized mice, while progression of the disease was observed in mice administered antigens in uncoated liposomes and those administered soluble antigens alone, as seen with control mice. Similarly, the number of parasites decreased substantially in local lymph nodes of mice immunized with the antigen in the Man5-DPPE-coated liposomes. Protection against L. major infection in the immunized mice also coincided with an elevated ratio of antigen-specific IgG2a/IgG1 antibodies, which is a profile of T helper-type 1-like immune response. Taken together, these results indicate the possibility that Man5-DPPE coated liposome-encapsulated antigens could serve as a vaccine that triggers protection against infectious disease. PMID- 12628647 TI - [3-(1H-imidazol-4-yl)propyl]guanidines containing furoxan moieties: a new class of H3-antagonists endowed with NO-donor properties. AB - Synthesis and pharmacological characterisation of a series of products obtained by coupling the H(3)-antagonist SKF 91486 through appropriate spacers with the NO donor 3-phenylfuroxan-4-yloxy and 3-benzenesulfonylfuroxan-4-yloxy moieties, as well as with the corresponding furazan substructures, devoid of NO-donating properties, are reported. All the products were tested for their H(3) antagonistic and H(2)-agonistic properties on electrically-stimulated guinea-pig ileum segments and guinea-pig papillary muscle, respectively. The whole series of compounds displayed good H(3)-antagonist behaviour and feeble partial H(2) agonist activity. Among furoxan derivatives, the benzenesulfonyl hybrid 28, a good NO-donor, triggered a dual NO-dependent muscle relaxation and H(3) antagonistic effect on guinea-pig intestine. PMID- 12628648 TI - Sugar derivatives as new 6-phosphogluconate dehydrogenase inhibitors selective for the parasite Trypanosoma brucei. AB - Sugar derivatives mimicking compounds which take part in the catalysed reaction have been assayed as alternative substrates and/or competitive inhibitors of 6 phosphogluconate dehydrogenase from Trypanosoma brucei and sheep liver. Phosphonate analogues have been synthesised and the new compound 5-deoxy-5 phosphono-D-arabinonate shows good selectivity towards the parasite enzyme. A number of 4-carbon and 5-carbon aldonates are strong inhibitors of the parasite enzyme with K(i) values below the substrate K(m) and some acyl derivatives are also potent inhibitors. At least five of the compounds showing a significant selectivity for the parasite enzyme represent leads for trypanocidal drugs against this recently validated target. PMID- 12628649 TI - Inhibition of protein kinase C by synthetic xanthone derivatives. AB - The modulatory activity of two xanthones (3,4-dihydroxyxanthone and 1-formyl-4 hydroxy-3-methoxyxanthone) on isoforms alpha, betaI, delta, eta and zeta of protein kinase C (PKC) was evaluated using an in vivo yeast phenotypic assay. Both xanthones caused an effect compatible with PKC inhibition, similar to that elicited by known PKC inhibitors (chelerythrine and NPC 15437). PKC inhibition caused by xanthones was confirmed using an in vitro kinase assay. The yeast phenotypic assay revealed that xanthones present differences on their potency towards the distinct PKC isoforms tested. It is concluded that 3,4 dihydroxyxanthone and 1-formyl-4-hydroxy-3-methoxyxanthone may become useful PKC inhibitors and xanthone derivatives can be explored to develop new isoform selective PKC inhibitors. PMID- 12628650 TI - An approach to identifying novel substrates of bacterial arylamine N acetyltransferases. AB - Arylamine N-acetyltransferases (NATs) catalyse the acetylation of arylamine, arylhydrazine and arylhydroxylamine substrates by acetyl Coenzyme A. NAT has been discovered in a wide range of eukaryotic and prokaryotic species. Although prokaryotic NATs have been implicated in xenobiotic metabolism, to date no endogenous role has been identified for the arylamine N-acetyl transfer reaction in prokaryotes. Investigating the substrate specificity of these enzymes is one approach to determining a possible endogenous role for prokaryotic NATs. We describe an accurate and efficient assay for NAT activity that is suitable for high-throughput screening of potential NAT ligands. This assay has been utilised to identify novel substrates for pure NAT from Salmonella typhimurium and Mycobacterium smegmatis which show a relationship between the lipophilicity of the arylamine and its activity as a substrate. The lipophilic structure/activity relationship observed is proposed to depend on the topology of the active site using docking studies of the crystal structures of these NAT isoenzymes. The evidence suggests an endogenous role of NAT in the protection of bacteria from aromatic and lipophilic toxins. PMID- 12628651 TI - New potent C2-symmetric malaria plasmepsin I and II inhibitors. AB - A series of malaria plasmepsin (Plm) I and II inhibitors containing a C(2) symmetric core structure have been synthesised and tested for protease inhibition activity. These compounds can be prepared using a straightforward synthesis involving a phenol nucleophilic ring opening of a diepoxide. Exemplar compounds synthesised exhibited remarkable inhibitory activity against both Plm I and II, notably 15c with K(i) values of 2.7nM and 0.25nM respectively, as well as showing >100-fold selectivity against Cathepsin D. PMID- 12628652 TI - Synthesis, binding affinity, and transcriptional activity of hydroxy- and methoxy substituted 3,4-diarylsalicylaldoximes on estrogen receptors alpha and beta. AB - An effective, unprecedented replacement of the prototypical phenolic 'A-ring' of estrogens with an oxime and a hydroxy-moiety of the salicylaldoxime derivative 3,4-diphenyl-substituted (1a) opened the way to study structure-activity relationships of a new class of estrogen receptor (ER)-ligands. Herein, we present a study of the ER binding properties and transcriptional activities of analogues of 3,4-diphenylsalicylaldoxime (1a). The introduction of p-OH and p-OMe groups on the phenyl substituents of 1a, as in compounds 1b-g, results in unique structure-activity profiles. The preparation of the hetero-disubstituted compounds (1b-e) was accomplished by a sequential introduction of different 3- and 4-aryl groups, obtained by exploiting the different reactivity of the bromine versus chlorine substituents on the precursor, 2-bromo-3-chloronitrobenzene (5), in the palladium-catalyzed cross-coupling reactions. The results of the biological tests show that the introduction of one hydroxy-group on the 3-phenyl substituent of the lead compound 1a improved the binding affinity on ERbeta (1c), whereas the introduction of the same group on the 4-phenyl substituent of 1a gave a compound (1e) with better affinity properties on ERalpha. The introduction of two hydroxyl groups in the para-position of both phenyl substituents of 1a, as in 1g, lowered the binding on both receptor subtypes. In transcription assays, the ERalpha agonist character of this class of ligands is enhanced by the presence of a p-hydroxy or p-methoxy in the 'distal' phenyl ring, whereas substitution on the other phenyl ring does not substantially modify the partial agonist character of 1a. Thus, results from the binding and transcription assays illustrate that this class of ER ligands has a distinct structure-activity profile on the two ER subtypes, being potent nearly full agonists on ERalpha and weak, partial antagonists on ERbeta. PMID- 12628653 TI - Synthesis and anticancer activity of 2-amino-8-chloro-5,5 dioxo[1,2,4]triazolo[2,3-b][1,4,2]benzodithiazine derivatives. AB - A new series of 1-(6-chloro-1,1-dioxo-1,4,2-benzodithiazin-3-yl)-4 arylsemicarbazides (4-16) were obtained. Intramolecular ring closure in semicarbazides 4-16 upon treatment with phosphorus oxychloride resulted in the formation of 2-amino-8-chloro-5,5-dioxo[1,2,4]triazolo[2,3 b][1,4,2]benzodithiazines 17-29 with potential antitumor activity. The structures of these compounds were confirmed on the basis of elemental analysis, spectral data and X-ray analysis. Compounds 17-29 were screened at the US National Cancer Institute (NCI) for their activities against a panel of 59 tumor cell lines, and relationships between structure and antitumor activity in vitro are discussed. The benzodithiazines 18, 19, 23, 28 and 29 were inactive, whereas the other compounds exhibited reasonable activity against numerous human tumor cell lines. The prominent compound 17 showed significant activity against the leukemia SR cell line (log GI(50)=-7.67, log TGI=-6.90 and log LC(50)=-4.77). PMID- 12628654 TI - Novel B-ring modified allocolchicinoids of the NCME series: design, synthesis, antimicrotubule activity and cytotoxicity. AB - Two new series of allocolchicinoids mimicking the structure of (-)-N acetylcolchinol O-methyl ether (2, NCME) were synthesized and evaluated for their abilities to inhibit tubulin assembly. Possible antitumor properties resulting thereof were evaluated in vitro on the human MCF-7 breast cancer cell line. The first series of NCME-derivatives was brought about by extending the seven membered B-ring to novel semisynthetic variations with a nitrogen containing eight-membered B-ring similar, for example, to the artificial, potent steganacin aza-analogue 3. In the second series the seven-membered B-ring of NCME (2) was modified by annulation with a heterocyclic ring system. The racemic ketone 7a serving as key precursor involved in the syntheses of all the target NCME variants 9-13 and 15, 16 was easily transformed into the eight-membered B-ring lactams 9 and 10 via a Beckmann rearrangement of the corresponding E-oxime 8. The tetrazole annulated congener 11 was prepared via azidotrimethylsilane-mediated Schmidt rearrangement. Treatment of educt 7a with Bredereck's reagent led to the enamino ketone 14, which was easily converted into the pyrazole- or pyrimidine annulated allocolchicinoids 15 and 16. Remarkably, all the allocolchicinoids 9-13 with an azocin-B-ring affected the tubulin/microtubule equilibrium only moderately. In contrast, the novel heterocycle annulated seven membered B-ring variants 15 and 16 proved to be highly potent tubulin-inhibitory, antimitotic agents. Interaction with tubulin occured at concentrations similar to those observed for colchicine (1) or the lead NCME (2). In all cases the antiproliferative effects correlated roughly with the inhibition of tubulin assembly. PMID- 12628656 TI - QSAR studies on piperazinylalkylisoxazole analogues selectively acting on dopamine D3 receptor by HQSAR and CoMFA. AB - QSAR studies for piperazinylalkylisoxazole analogues were conducted by hologram QSAR (HQSAR) and comparative molecular field analysis (CoMFA) to explain the binding affinities of 264 ligands acting on dopamine D(3) receptor. The HQSAR was assessed by r(2) value of 0.917 and cross validated q(2) value of 0.841. In the CoMFA, r(2) is 0.919 and cross validated q(2) is 0.727. The results provide the tools for predicting the affinity of related compounds and guiding the design of new ligands. PMID- 12628655 TI - Synthesis and analgesic activity of a series of new azaalkane bis-guanidinium and bis(2-aminoimidazolinium) compounds. AB - In the present paper, we wish to report the synthesis and antinociceptive activity of a series of new azaalkane bis(2-aminoimidazolinium) compounds from which, N,N'-di(4,5-dihydro-1H-imidazol-2-yl)-3-aza-1,6-hexanediamine 2a has shown the best analgesic properties in vivo in two different assays (i.e., acetic acid induced writhing test and hot-plate test in mice), as well as oral bioavailability. PMID- 12628657 TI - Inhibition of monoamine oxidase B by selective adenosine A2A receptor antagonists. AB - Adenosine receptor antagonists that are selective for the A(2A) receptor subtype (A(2A) antagonists) are under investigation as possible therapeutic agents for the symptomatic treatment of the motor deficits associated with Parkinson's disease (PD). Results of recent studies in the MPTP mouse model of PD suggest that A(2A) antagonists may possess neuroprotective properties. Since monoamine oxidase B (MAO-B) inhibitors also enhance motor function and reduce MPTP neurotoxicity, we have examined the MAO-B inhibiting properties of several A(2A) antagonists and structurally related compounds in an effort to determine if inhibition of MAO-B may contribute to the observed neuroprotection. The results of these studies have established that all of the (E)-8-styrylxanthinyl derived A(2A) antagonists examined display significant MAO-B inhibitory properties in vitro with K(i) values in the low micro M to nM range. Included in this series is (E)-1,3-diethyl-8-(3,4-dimethoxystyryl)-7-methylxanthine (KW-6002), a potent A(2A) antagonist and neuroprotective agent that is in clinical trials. The results of these studies suggest that MAO-B inhibition may contribute to the neuroprotective potential of A(2A) receptor antagonists such as KW-6002 and open the possibility of designing dual targeting drugs that may have enhanced therapeutic potential in the treatment of PD. PMID- 12628658 TI - 4-Hydroxymethyl- and 4-methoxymethylfuro[2,3-h]quinolin-2(1H)-ones: synthesis and biological properties. AB - 4-Hydroxymethyl-1,6,8-trimethylfuro[2,3-h]quinolin-2(1H)-one (HOFQ) was prepared by a new profitable way, which allowed to synthesize also 4-methoxymethyl-1,6,8 trimethylfuro[2,3-h]quinolin-2(1H)-one (MOFQ), and 4-hydroxymethyl-6,8 dimethylfuro[2,3-h]quinolin-2(1H)-one (HOHFQ). Some biological activities of the three compounds were studied in comparison with 8-MOP. In the dark, they inhibited topoisomerase II, leading to a moderate antiproliferative activity in mammalian cells. The antiproliferative activity was also tested upon UVA irradiation in mammalian cells: all compounds showed higher activity than 8-MOP, without mutagenicity and skin phototoxicity, with the best results for HOFQ. Photobinding to DNA was investigated, demonstrating a different sequence specificity for these furoquinolinones in comparison with furocoumarins. For all these features, HOFQ and the other analogues appeared very promising photochemotherapeutic agents, whose mechanism of action will be further investigated. PMID- 12628659 TI - Design and synthesis of 1,5- and 2,5-substituted tetrahydrobenzazepinones as novel potent and selective integrin alphaVbeta3 antagonists. AB - The design and synthesis of novel integrin alpha(V)beta(3) antagonists based on a 1,5- or 2,5-substituted tetrahydrobenzaezpinone core is described. In vitro activity of respective compounds was determined via alpha(V)beta(3) binding assay, and selected derivatives were submitted to further characterization in functional cellular assays. SAR was obtained by modification of the benzazepinone core, variation of the spacer linking guanidine moiety and core, and modification of the guanidine mimetic. These efforts led to the identification of novel alpha(V)beta(3) inhibitors displaying potency in the subnanomolar range, selectivity versus alpha(IIb)beta(3) and functional efficacy in relevant cellular assays. A method for the preparation of enantiomerically pure derivatives was developed, and respective enantiomers evaluated in vitro. Compounds 31 and 37 were assessed for metabolic stability, resorption in the Caco-2 assay and pharmacokinetics. PMID- 12628660 TI - Evaluation of dipeptide-derivatives of 5-aminolevulinic acid as precursors for photosensitizers in photodynamic therapy. AB - N-terminal-blocked and N-terminal-free pseudotripeptide Gly-Gly and Gly-Pro derivatives of 5-aminolevulinic acid (ALA) esters were synthesized as potential specific substrates for cellular peptidases and precursors for the production of the photosensitizer protoporphyrin IX (PpIX). These precursors were evaluated using human cell lines of either carcinoma or endothelial origin. N-blocked or N free dipeptides-ALA-ethyl esters, but not tripeptides-ALA-ethyl esters (or dipeptides-ALA-ethyleneglycols,) were substrates for cellular peptidases and were metabolized to ALA. The precursors were hydrolyzed intracellularly involving serine-proteases and metalloproteases. Cell selectivity for human endothelial or carcinoma cells was observed for some of these dipeptides-ALA. Thus drugs coupled to Gly-Gly-/Gly-Pro-derivatives may selectively target defined cells in human cancer, depending on specific cellular activating pathways expressed by the cells. PMID- 12628661 TI - Cardiovascular hybrid drugs: new benzazepinone derivatives as bradycardic agents endowed with selective beta1-non-competitive antagonism. AB - The synthesis and pharmacological profile of some hybrid compounds bearing both the benzazepinone moiety present in Zatebradine and typical beta-blocker aryloxypropanolamine groups are described. The new compounds proved to be endowed with negative chronotropic and inotropic activity and are weak vasorelaxant agents. The cardiodepressant action is probably due to selective beta(1) noncompetitive reversible antagonism. Both enantiomers of the most active compound 5c were synthesized and they showed a different cardiovascular profile, that is (+)-(R)-enantiomer displays affinity for cardiac beta(1)-adrenoceptors, while (-)-(S)-enantiomer shows specificity for vessel smooth muscle. PMID- 12628662 TI - Complementary characteristics of homologous p-octiphenyl beta-barrels with ion channel and esterase activity. AB - We report that decreasing beta-sheet length in homologous multifunctional rigid rod beta-barrels with internal histidines increases ion channel stability by three orders of magnitude, reduces binding activity by four orders of magnitude, and reduces esterase activity up to 22-times. These results are further used to evaluate methods employed to characterize suprastructure and activity of synthetic multifunctional pores formed by p-octiphenyl beta-barrels with emphasis on applicability of the Hille model to determine internal diameters and the Woodhull equation to locate internal active sites. PMID- 12628663 TI - Exploration of cornea permeable calpain inhibitors as anticataract agents. AB - To explore cornea permeable calpain inhibitors, four compounds displaying different characteristics were designed and synthesized based on the known potent calpain inhibitor, peptidyl aldehyde SJA6017. Two approaches were adopted; an improvement in the physicochemical properties, and conversion of the active aldehyde. The water-soluble peptidyl aldehyde 1 containing a pyridine ring at the P3 site showed a modest inhibition against calpains and an improvement of corneal permeability in comparison with SJA6017. Replacement of the aldehyde of SJA6017 by an alpha-ketoamide provided compound 2 that was approximately equipotent with SJA6017, but it was extremely water-insoluble. However, compound 3, in which the aldehyde was converted into a cyclic hemiacetal, proved to be a less potent calpain inhibitor than SJA6017, but demonstrated excellent transcorneal permeability. Further modification generating the cyclic hemiacetal 4 containing a thiourea linker between the P3 and P2 sites exhibited potent inhibitory activities, high cornea permeability and excellent efficacy in the rat lens culture cataract model. PMID- 12628664 TI - Synthesis and in vitro leishmanicidal activity of some hydrazides and their analogues. AB - Twenty-one hydrazides were synthesized by treating different esters with hydrazine hydrate. Substituted hydrazides were obtained by treating hydrazides with alkyl/aryl/acyl halides. Some of these compounds exhibit potential in vitro leishmanicidal activity. The structures of all the synthesized compounds were confirmed by spectroscopic analysis. PMID- 12628665 TI - De novo design, synthesis and evaluation of a non-steroidal diphenylnaphthyl propylene ligand for the estrogen receptor. AB - There is still a strong need for additional diversity and new chemical scaffolds to allow for the exploration of improved tissue selectivity and finding better selective estrogen receptor modulators (SERMs). Using a de novo design technology a diphenylnaphthyl propylene scaffold, exemplified by (E)-9b, with ER antagonist activity has been generated. It was prepared by alkylating 1-[4-methoxyphenyl)-2 (4-(2-chloroethoxy)phenyl]-1-propanone under metal halogen exchange conditions with 1-iodo-6-methoxy-naphthalene. Following dehydration and cleavage of the methoxy groups, (E)-9b was formed by displacement of the chloro group with pyrrolidine. (E)-9b binding to ER generated calculated K(i) values of 3.7 nM for hER(alpha) and 72 nM for hER(beta). The antagonism of (E)-9b was demonstrated in cell transfection assays using the ERE from the vitA2 promotor and the natural ER responsive pS2 promotor. With increasing concentrations of (E)-9b, the E(2) dependent response was efficiently inhibited demonstrating that (E)-9b could function as an anti-estrogen in these assays. Interestingly, ER(alpha) activity was inhibited even below basal levels suggesting that ligand-independent activity of ER(alpha) was also inhibited. Computational docking studies suggest that the placement of the hydroxyl group on the naphthalene group may not be optimal and we are currently exploring additional analogues. PMID- 12628667 TI - Oxidative deamination of benzylamine by glycoxidation. AB - In the present study, model reactions for the oxidative deamination by glycoxidation using benzylamine were undertaken to elucidate the detail of the reaction. Glucose, 3-deoxyglucosone (3-DG), and methylglyoxal (MG) oxidatively deaminated benzylamine to benzaldehyde in the presence of Cu(2+) at a physiological pH and temperature but not glyoxal. 3-DG and MG were more effective oxidants than glucose. We have determined the effects of metal ions, pH, oxygen, and radical scavengers on the oxidative deamination. The formation of benzaldehyde was greatest with Cu(2+), and was accelerated at a higher pH and in the presence of oxygen. EDTA, catalase, and dimethyl sulfoxide significantly inhibited the oxidation, suggesting the participation of reactive oxygen species. From these results, we propose a mechanism for the oxidative deamination by the Strecker-type reaction and the reactive oxygen species-mediated oxidation during glycoxidation. PMID- 12628666 TI - Structure-activity relationships for 1',1'-dimethylalkyl-Delta8 tetrahydrocannabinols. AB - A series of 1',1'-dimethylalkyl-Delta(8)-tetrahydrocannabinol analogues with C-3 side chains of 2-12 carbon atoms has been synthesized and their in vitro and in vivo pharmacology has been evaluated. The lowest member of the series, 1',1' dimethylethyl-Delta(8)-THC (8, n=0) has good affinity for the CB(1) receptor, but is inactive in vivo. The dimethylpropyl (8, n=1) through dimethyldecyl (8, n=8) all have high affinity for the CB(1) receptor and are full agonists in vivo. 1',1'-Dimethylundecyl-Delta(8)-THC (8, n=9) has significant affinity for the receptor (K(i)=25.8+/-5.8 nM), but has reduced potency in vivo. The dodecyl analogue (8, n=10) has little affinity for the CB(1) receptor and is inactive in vivo. A quantitative structure-activity relationship study of the side chain region of these compounds is consistent with the concept that for optimum affinity and potency the side chain must be of a length which will permit its terminus to loop back in proximity to the phenolic ring of the cannabinoid. PMID- 12628668 TI - An efficient oxidizing reagent for the synthesis of mixed backbone oligonucleotides via the H-phosphonate approach. AB - The mixture of carbon tetrachloride, N-methyl morpholine (NMM), pyridine and water in acetonitrile has been exploited for the oxidation of dinucleoside H phosphonate diesters to the corresponding phosphates. The system is found to be inert to the phosphoramidate (P-N) and the phosphorothioate (P-S) linkages and has successfully been applied to the solid phase synthesis of mixed-backbone oligonucleotides (MBOs). PMID- 12628669 TI - Respiratory chain inhibition by fullerene derivatives: hydrogen peroxide production caused by fullerene derivatives and a respiratory chain system. AB - Fullerene is a new type of carbon allotrope. We have shown that the fullerene derivative C(60)-bis(N,N-dimethylpyrrolidinium iodide), a regio isomer mixture, inhibited Escherichia coli growth and dioxygen uptake caused by E. coli and glucose. This result indicates that the mechanism of the bacteriostatic effect is the inhibition of energy metabolism. In this study, we isolated two regio isomers of C(60)-bis(N,N-dimethylpyrrolidinium iodide) and studied their effect on E. coli growth and on respiratory chain activity. In dioxygen uptake caused by the inner-membrane and NADH, the effect of fullerene derivatives was biphasic. At low concentrations of both fullerene derivatives, dioxygen uptake was inhibited, whereas at high concentrations, it was increased. At high concentrations, consumed dioxygen was converted to H(2)O(2). An electrochemical study revealed that reduced fullerene derivatives react with dioxygen. This activity was closely related to a redox property of the isomers. PMID- 12628670 TI - Investigation of platelet aggregation inhibitory activity by phenyl amides and esters of piperidinecarboxylic acids. AB - A series of anilides and phenyl esters of piperidine-3-carboxylic acid (nipecotic acid) were synthesized and tested for the ability to inhibit aggregation of human platelet rich-plasma triggered by adenosine 5'-diphosphate (ADP) and adrenaline. As a rule, amides were about two times more active than the corresponding esters, and derivatives bearing substituents at the para position of the phenyl ring were significantly more active than the meta-substituted ones. Among the tested compounds, 4-hexyloxyanilide of nipecotic acid (18a) was found to be the most active one, its IC(50) value being close to that of the most active bis-3 carbamoylpiperidines reported in literature (ca. 40 micro M) and aspirin (ca. 60 microM) in ADP- and adrenaline-induced aggregation, respectively. Compared with the isomeric 4-hexyloxyanilides of piperidine-2-carboxylic (pipecolinic) and piperidine-4-carboxylic (isonipecotic) acids, compound 18a showed higher activity, and a Hansch-type quantitative structure-activity relationship (QSAR) study highlighted lipophilicity and increase in electron density of the phenyl ring as the properties which mainly increase the antiplatelet activity (r(2)=0.74, q(2)=0.64). The interaction of nipecotoyl anilides with phosphatidylinositol, a major component of the inner layer of the platelet membranes, was investigated by means of flexible docking calculation methods to give an account of a key event underlying their biological action. PMID- 12628672 TI - Study on molecular mechanism and 3D-QSAR of influenza neuraminidase inhibitors. AB - Neuraminidase (NA) is a critical enzyme of the influenza virus and many inhibitors targeting to this enzyme are quite efficient and encouraging as anti influenza agents. In this paper the binding model of five series of inhibitors to NA was examined using molecular simulation method. The resulted conformation and orientation of the compounds were directly put into CoMSIA study. The most significant amino acid residues at binding sites and the requirement for features of substituents were applied to direct design of new inhibitors. The robust QSAR model and its three-dimensional contour map provided guidelines to building novel compounds with new scaffold and for structural optimization of current molecules. PMID- 12628671 TI - Synthesis and pharmacological evaluation of substituted naphth[1,2,3 de]isoquinolines (dinapsoline analogues) as D1 and D2 dopamine receptor ligands. AB - Dinapsoline ((2); (+/-)-dihydroxy-2,3,7,11b-tetrahydro-1H-naphth[1,2,3 de]isoquinoline) is a full D(1) dopamine agonist that also has significant D(2) receptor affinity. Based on a similar pharmacophore, dinapsoline has pharmacological similarities to dihydrexidine ((1); (+/-)-trans-10,11-dihydroxy 5,6,6a,7,8,12b-hexahydrobenzo[a]phenanthridine), the first high affinity full D(1) agonist. Small alkyl substitutions on the dihydrexidine backbone are known to alter markedly the D(1):D(2) selectivity of dihydrexidine, and it was of interest to determine whether similar SAR exists within the dinapsoline series. This report describes the synthesis and pharmacological evaluation of six analogues of dinapsoline: N-allyl-(3);N-n-propyl- (4); 6-methyl- (5); 4-methyl- (6); 4-methyl-N-allyl- (7); and 4-methyl-N-n-propyl-dinapsoline (8). As expected from earlier studies with the dihydrexidine backbone, N-allyl (3) or N-n-propyl (4) analogues had markedly decreased D(1) affinity. Unexpectedly, and unlike the dihydrexidine series, these same substituents did not markedly increase D(2) affinity. The addition of a methyl group to position 6 (5) increased D(1):D(2) selectivity, but less markedly than did the analogous 2-methyl substituent added to 1. Unlike the analogous 4-methyl substituent of 1, the addition of a 4-methyl group (6) actually decreased D(1) affinity without affecting D(2) affinity. These data demonstrate that the dinapsoline (2) backbone can be modified to produce dopamine agonists with novel properties. Moreover, as rigid ligands in which small substituents can cause significant changes in selectivity, they are important tools for deriving 'differential' SARs of the dopamine receptor isoforms. PMID- 12628673 TI - Substituted dibenzo[c,h]cinnolines: topoisomerase I-targeting anticancer agents. AB - Several substituted dibenzo[c,h]cinnolines were synthesized and evaluated for their potential to target topoisomerase I and for their relative cytotoxic activity. Select benzo[i]phenanthridines are capable of stabilizing the cleavable complex formed with topoisomerase I and DNA. This study was initiated to examine whether dibenzo[c,h]cinnolines, which are in essence aza analogues of benzo[i]phenanthridines, possess similar pharmacological properties. 2,3 Dimethoxy-8,9-methylenedioxybenzo[i]phenanthridine is one of the more potent benzo[i]phenanthridine derivatives in regard to topoisomerase I-targeting activity and cytotoxicity. The structure-activity relationship observed with these substituted dibenzo[c,h]cinnolines parallels that observed for benzo[i]phenanthridine derivatives. Compared to similarly substituted benzo[i]phenanthridines, the dibenzo[c,h]cinnoline analogues exhibit more potent topoisomerase I-targeting activity and cytotoxicity. The relative IC(50) values obtained in assessing the cytotoxicity of 2,3-dimethoxy-8,9 methylenedioxydibenzo[c,h]cinnoline and 2,3-dimethoxy-8,9 methylenedioxybenzo[i]phenanthridine in the human lymphoblastma cell line, RPMI8402, are 70 and 400 nM, respectively. In tumor cell lines selected for resistance to camptothecin and known to express mutant topoisomerase I, benzo[i]phenanthridine derivatives were not cross-resistant. In contrast, similarly substituted dibenzo[c,h]cinnolines with significant topoisomerase I targeting activity did exhibit cross-resistance in these camptothecin-resistant cell lines. The cytotoxicity of these dibenzo[c,h]cinnolines was not diminished in cells overexpressing the efflux transporter, MDR1. These data indicate that substituted dibenzo[c,h]cinnolines can exhibit potent topoisomerase I-targeting activity and are capable of overcoming the multi-drug resistance associated with this efflux transporter. PMID- 12628674 TI - New thiazole derivatives as potent and selective 5-hydroxytriptamine 3 (5-HT3) receptor agonists for the treatment of constipation. AB - The syntheses and biological evaluation of a series of novel indeno[1,2 d]thiazole derivatives are described. Several groups reported 5-HT(3) receptor agonists which were mainly evaluated for their activities on the von Bezold Jarisch reflex (B-J reflex). We discovered that tetrahydrothiazolopyridine derivative 1b had a contractile effect on the isolated guinea pig colon with weak B-J reflex. Our efforts to find a new type of 5-HT(3) receptor agonists on the isolated guinea pig colon focused on the synthesis of a fused thiazole derivative 1d modified from 1b and reverse-fused thiazole derivatives (7-10). In this series, 10f (YM-31636) showed high affinity and selectivity for the cloned human 5-HT(3) receptor; furthermore, it showed potent and selective 5-HT(3) receptor agonistic activity. YM-31636 was examined for its effects on defecation in animals, thus evaluating the compound as an agent against constipation. PMID- 12628675 TI - Parthenolide and its photochemically synthesized 1(10)Z isomer: chemical reactivity and structure-activity relationship studies in human leucocyte chemotaxis. AB - The present study has achieved the photochemical conversion of a germacrolide into a melampolide. The investigation on their chemical properties allowed us to evaluate the minimum interatomic distance needed for transannular bridging of C(10) ring in germacrolides and to explain the regiochemical selectivity of electrophilic cyclizations. The antiinflammatory activity of parthenolide and its semisynthetic derivatives was evaluated by in vitro chemotaxis assay with human neutrophiles. These structure-activity relationship studies have led to hypothesize a new pharmacophore and have provided useful information for computationally designed drugs. PMID- 12628676 TI - Synthesis and cytotoxicity evaluation of 2-amino- and 2-hidroxy-3-ethoxycarbonyl N-substituted-benzo[f]indole-4,9-dione derivatives. AB - Reaction between 2,3-dichloronaphthoquinone (I) and ethyl cyanoacetate or diethyl malonate under different conditions gave the starting materials, 2-chloro-3 (alpha-cyano-alpha-ethoxycarbonyl-methyl)-1,4-naphthoquinone (A) or 2-chloro-3 (diethoxycarbonyl-methyl)-1,4-naphthoquinone (B). The 2-amino-3-ethoxycarbonyl-N substituted-benzo[f]indole-4,9-dione derivatives [A-(1-10)] and 2-hydroxy-3 ethoxycarbonyl-N-substituted-benzo[f]indole-4,9-dione derivatives [B-(1-12)] were prepared from compounds A and B, respectively, by using various alkyl-, and arylamines. The cytotoxic activities of the prepared compounds were evaluated by SRB (Sulforhodamine B) assay against the following tumor cell lines: A459 (human lung), SK-OV-3 (human ovarian), SK-MEL-2 (human melanoma), XF498 (human CNS), and HCT 15 (human colon). Many of the derivatives mentioned exhibited more potent cytotoxic effects against SK-OV-3 and XF498 than etoposide. Significantly, 2 amino-3-ethoxycarbonyl-N-(3-methyl-phenyl)-benzo[f]indole-4,9-dione (A-8) showed potent activity against all tumor cell lines, and in particular, its cytotoxic effect against SK-OV-3 was much higher than doxorubicin. PMID- 12628677 TI - In vitro antifungal activity of new series of homoallylamines and related compounds with inhibitory properties of the synthesis of fungal cell wall polymers. AB - The synthesis, in vitro antifungal evaluation and SAR studies of 101 compounds of the 4-aryl-, 4-alkyl-, 4-pyridyl or -quinolinyl-4-N-arylamino-1-butenes series and related compounds, are reported here. Active structures showed to inhibit (1,3)-beta-D-glucan and mainly chitin synthases, enzymes that catalyze the synthesis of the major fungal cell wall polymers. PMID- 12628678 TI - Semi-synthesis of an O-glycosylated docetaxel analogue. AB - A 7beta-O-glycosylated docetaxel analogue was semi-synthesized from 9-dihydro-13 acetylbaccatin III, the most abundant taxane isolated from the needles of Taxus canadensis. It was shown to be more bioactive than paclitaxel according to the tubulin assay. It had a reduced potency in the MCF7 cell line cytotoxicity assay compared to paclitaxel, but it demonstrated better activity against the drug resistant cell line MCF7-ADR. In addition, the presence of one sugar moiety on C 7 doubled the water solubility versus that of paclitaxel. PMID- 12628679 TI - Synthesis and biological evaluation of C-3'NH/C-10 and C-2/C-10 modified paclitaxel analogues. AB - Concurrent modifications on the C-3'NH/C-10, and C-2/C-10 positions on paclitaxel were carried out as a way of investigating possible synergistic effects. The biological activities of these analogues were evaluated in both a microtubule assembly assay and human ovarian cancer (A2780) and prostate cancer (PC3) cytotoxicity assay. In some cases the doubly modified analogues were more active than would have been predicted based on the activity of the singly modified analogues, indicating probable synergistic effects. PMID- 12628680 TI - Synthesis and preliminary cytotoxicity study of glucuronide derivatives of CC 1065 analogues. AB - Glucuronide derivatives of CBI-bearing CC-1065 analogues have been synthesized, and their cytotoxicities tested against U937 leukemia cells. The new compounds show potent antitumor activity in vitro. Compounds 1 and 2, and their corresponding glucuronides 3 and 4 have IC(50) values of 0.6, 0.1, 1.4 and 0.6 nM, respectively. Glucuronide 3 is approximately 2-fold less toxic than its hydroxyl counterpart 1, and glucuronide 4 is approximately 6-fold less toxic than its hydroxyl counterpart 2. Glucuronides 3 and 4 may have limited use in the ADEPT approach. However, they may be used as antitumor agents in a conventional way. PMID- 12628681 TI - Integracides: tetracyclic triterpenoid inhibitors of HIV-1 integrase produced by Fusarium sp. AB - HIV-1 integrase is a critical enzyme in the replication of HIV-1. It is absent in the host cells and therefore is a good target for treatment of HIV-1 infections. Integracides are members of the tetracyclic triterpenoids family that were isolated from the fermentation broth of a Fusarium sp. Integracide A, a sulfated ester, exhibited significant inhibitory activity against strand transfer reaction of HIV-1 integrase. The discovery, structure elucidation including single crystal X-ray structure and HIV-1 inhibitory activity of these compounds are described. PMID- 12628682 TI - Combinatorial enzymatic assay for the screening of a new class of bacterial cell wall inhibitors. AB - We have developed a screening assay by thin-layer chromatography (TLC) to identify inhibitors for the bacterial essential enzymes MurA, -B, and -C. Libraries of compounds were synthesized using the mix-and-split combinatorial chemistry approach. Screening of the pooled compounds using the developed assay revealed the presence of many pools active in vitro. Pools of interest were tested for antibacterial activity. Individual molecules in the active pools were synthesized and retested with the TLC assay and with bacteria. We focused on the best five compounds for further analysis. They were tested for inhibition on each of the three enzymes separately, and showed no inhibition of MurA or MurB activity but were all inhibitors of MurC enzyme. This approach yielded interesting lead compounds for the development of novel antibacterial agents. PMID- 12628683 TI - DNA damaging activity of ellagic acid derivatives. AB - A strain of yeast rendered repair deficient by the conditional expression of the RAD52 locus was used to search for natural products capable of damaging DNA. Four ellagic acid derivatives, namely 3,3'-dimethyl-4'-O-beta-D-glucopyranosyl ellagic acid (1), 3,3',4-trimethyl-4'-O-beta-D-glucopyranosyl ellagic acid (2), 3'-methyl 3,4-O,O-methylidene ellagic acid (3) and 3'-methyl-3,4-O,O-methylidene-4'-O-beta D-glucopyranosyl ellagic acid (4), were identified by this assay as DNA damaging natural principles from several plants, including Alangium javanicum, Anisophyllea apetala, Crypteronia paniculata, Mouririi sp. and Scholtzia parviflora. Although none of the isolated principles mediated frank strand scission of DNA in vitro, all of them potently inhibited the growth of yeast in the absence of expression of RAD52. PMID- 12628684 TI - Structure-activity relationship study of human liver microsomes-catalyzed hydrolysis rate of ester prodrugs of MENT by comparative molecular field analysis (CoMFA). AB - A series of MENT esters (3-71) was designed, prepared and tested to study the structure-activity relationship (SAR) of the hydrolysis rate with human liver microsomes of these prodrugs. Compounds were obtained covering a wide range of metabolic stability. The results are useful for the proper selection of prodrugs for different pharmaceutical formulations to deliver the potent and prostate sparing androgen MENT. The MENT esters can especially be administered for male hormone replacement therapy and male contraception. Comparative molecular field analysis (CoMFA) was applied to a dataset of 28 esters, for which ED50 values could be obtained. The CoMFA model where the electrostatic and H-bond molecular fields were combined turned out to be most predictive. Despite the limited size of the dataset, CoMFA can help to rationalize the SAR of the ester hydrolysis rate of ester prodrugs of MENT. PMID- 12628685 TI - Chromatographic behavior of oxygenated derivatives of cholesterol. AB - Oxygenated derivatives of cholesterol have important functions in many biochemical processes. These oxysterols are difficult to study because of their low physiological concentrations, the facile formation of cholesterol autoxidation artifacts, and lack of information on their chromatographic behavior. Focusing on metabolites and autoxidation products of cholesterol, we have documented the chromatographic mobilities of 35 oxysterols under a variety of conditions: eight solvent systems for thin-layer chromatography on silica gel, several mobile phases for reversed-phase high-performance liquid chromatography (HPLC), and two types of stationary phase for capillary gas chromatography (GC) using trimethylsilyl derivatives. Notable differences in selectivity could be obtained by modifying the stationary or mobile phases. Separations of oxysterol pairs isomeric at side-chain carbons or C-7 were achieved on normal-phase, reversed-phase, chiral, or silver-ion HPLC columns. Chromatographic behavior is also described for side-chain hexadeuterated and heptafluorinated oxysterols, which are useful as standards in isotope dilution analyses and autoxidation studies, respectively. The overall results are relevant to many problems of oxysterol analysis, including the initial separation of oxysterols from cholesterol, determination of highly polar and nonpolar oxysterols, separation of isomeric pairs, selection of derivatization conditions for GC analysis, and quantitation of the extent of cholesterol autoxidation. PMID- 12628686 TI - Tibolone is not converted by human aromatase to 7alpha-methyl-17alpha ethynylestradiol (7alpha-MEE): analyses with sensitive bioassays for estrogens and androgens and with LC-MSMS. AB - To exclude that aromatization plays a role in the estrogenic activity of tibolone, we studied the effect tibolone and metabolites on the aromatization of androstenedione and the aromatization of tibolone and its metabolites to 7alpha methyl-17alpha-ethynylestradiol (7alpha-MEE) by human recombinant aromatase. Testosterone (T), 17alpha-methyltestosterone (MT), 19-nortestosterone (Nan), 7alpha-methyl-19-nortestosterone (MENT) and norethisterone (NET) were used as reference compounds. Sensitive in vitro bioassays with steroid receptors were used to monitor the generation of product and the reduction of substrate. LC-MSMS without derivatization was used for structural confirmation. A 10 times excess of tibolone and its metabolites did not inhibit the conversion of androstenedione to estrone by human recombinant aromatase as determined by estradiol receptor assay whereas T, MT, Nan, and MENT inhibited the conversion for 75, 53, 85 and 67%, respectively. Tibolone, 3alpha- and 3beta-hydroxytibolone were not converted by human aromatase whereas the estrogenic activity formed with the Delta4-isomer suggests a conversion rate of 0.2% after 120 min incubation. In contrast T, MT, Nan, and MENT were completely converted to their A-ring aromates within 15 min while NET could not be aromatized. Aromatization of T, MT, Nan and MENT was confirmed with LC-MSMS. Structure/function analysis indicated that the 17alpha ethynyl-group prevents aromatization of (19-nor)steroids while 7alpha-methyl substitution had no effect. Our results with the sensitive estradiol receptor assays show that in contrast to reference compounds tibolone and its metabolites are not aromatized. PMID- 12628688 TI - Conversion of cholic acid to methyl 3alpha-carbethoxy-12alpha-acetoxy-6-oxo-5beta chol-7-en-24-oate. AB - A new and efficient method for preparation of a 7-en-6-one derivative of cholic acid is described. Acetylation of the known methyl 3alpha-carbethoxy-12alpha hydroxy-7-oxo-5beta-cholan-24-oate (3) at 12 position and reduction of its 7-oxo group yield the 12alpha-acetoxy-7alpha-hydroxy derivative 5. Dehydration of the 7alpha-hydroxy group in 5 followed by allylic oxidation provide methyl 3alpha carbethoxy-12alpha-acetoxy-6-oxo-5beta-chol-7-en-24-oate (7) in good yield. PMID- 12628687 TI - Mouse Leydig tumor cells produce C-19 steroids, including testosterone. AB - The mouse Leydig tumor cells (MLTC-1) were derived from a transplantable Leydig cell tumor carried in C57BL/6 mice. The original cell line (M5480) produced testosterone and little progesterone. However, it was later shown that there were two subtypes of the cell line, one producing mainly progesterone and termed M5480P and the other which produced androgens and termed M5480A. MLTC-1 cells are reportedly derived from the former. We studied the production of testosterone by MLTC-1 cells using a specific and sensitive testosterone RIA, tandem mass spectrometry (TMS) and examined the expression of mRNA of some key enzymes involved in steroidogenesis. Although the molar yields were 1:20:60 for testosterone, androstenedione and progesterone, respectively, in response to human chorionic gonadotropin (hCG), testosterone measured by our RIA accounted for 94% of the testosterone immunoreactivity. Both MLTC-1 and Balb/c Leydig cells expressed Steroidogenic Acute Response (StAR) protein mRNA in response to hCG. Cytochrome P450 17alpha-hydroxylase/17,20-lyase mRNA was expressed constitutively in MLTC-1 and Balb/c Leydig cells. Whereas the latter expressed 17beta hydroxydehydrogenase/17-ketoreductase isoform Type 3mRNA in response to hCG, MLTC 1 cells expressed isoform Type 7 constitutively. The absence of isoform Type 3 in MLTC-1 cells thus may account for the low conversion of androstenedione to testosterone in this cell line. However, with a very specific and sensitive RIA even the low production of testosterone becomes meaningful. In conclusion MLTC-1 cells produce testosterone. PMID- 12628690 TI - Effect of a novel steroid (PM-9) on the inhibition of 5alpha-reductase present in Penicillium crustosum broths. AB - The conversion of testosterone (T) to 5alpha-dihydrotestosterone (DHT) has been demonstrated in Penicillium crustosum broth obtained from fermented pistachios, lemons and corn tortillas. Furthermore, the presence of 5alpha-reductase enzyme, which is responsible for this conversion, has been established by electrophoretical techniques in these cultures.5alpha-Reductase enzyme is also present in animal and human androgen-dependent tissues as well as in prostate and seminal vesicles. The increase of the conversion of T to DHT in prostate gland, has been related to some illnesses such as benign prostate hyperplasia and prostate cancer. Furthermore, treatment with 5alpha-reductase inhibitors such as finasteride reduces the prostate growth. These data have stimulated research for the synthesis of new molecules with antiandrogenic activity, whose biological effect needs to be demonstrated. The purpose of this study is to determine the inhibition pattern of 5alpha-reductase in P. crustosum by finasteride and the new steroidal compound PM-9. K(m) and V(max) values for T, were determined in the broths by Lineweaver-Burk plots using different testosterone concentrations. The inhibition pattern of finasteride and PM-9 was also determined by Lineweaver-Burk using different concentrations of T and inhibitors. Results show that finasteride and PM-9 inhibit 5alpha-reductase present in the broth in a competitive manner. PMID- 12628689 TI - Decrease of estrogen receptor expression and associated ERE-dependent transcription in MCF-7 breast cancer cells after oligomycin treatment. AB - Oligomycin at 0.01 microM produces very rapid decrease of [3H]estradiol (E2) binding capacity in MCF-7 cells maintained in culture in glucose- and serum-free medium. Loss of binding capacity was associated with elimination of the estrogen receptor (ER) as well as a decrease of basal expression of ERE-luciferase reporter gene. These effects were not due to major cell death as shown by MTT assay. Hence, the inhibition of ATP synthesis produced by oligomycin seems to influence ER turnover, resulting in very rapid loss of receptor. Withdrawal of oligomycin and maintenance of glucose in the medium led to only a partial reappearance of ER and failed to restore optimal ERE-dependent transcription. Oligomycin significantly down-regulated progesterone receptor (PR) level and partially abrogated E2-induced PR up-regulation, indicating that this drug also affects other nuclear receptors. Treatment of cytosol from MCF-7 cells with acid and alkaline phosphatases decreased [3H]E2-binding capacity, indicating the requirement of ER phosphorylation for optimal hormone binding. On the other hand, oligomycin-induced ER loss was partly compensated by E2 and partial anti estrogens (AEs; 4-OH-TAM or RU 39 411); i.e. oligomycin failed to improve the E2 induced ER down-regulation and very weakly suppressed partial AE-induced receptor up-regulation. The known ability of these ligands to stabilize ER in the cell nucleus before regulating ER level may explain this phenomenon since such antagonism was not recorded with pure AE RU 58 668, which is known to impede nuclear translocation of the receptor. Interestingly, ligands able to down regulate ER (i.e. E2 or RU 58 668) increased ER phosphorylation while 4-OH-TAM which up-regulate the receptor had little effect in this regard. Oligomycin failed to strongly affect such phosphorylation enhancements while it produced a weak decrease of basal phosphorylation level. Hence, phosphorylations/dephosphorylations of specific sites on ER and/or co-regulators seem to govern its turnover. PMID- 12628691 TI - Synthesis and receptor-binding examinations of the normal and 13-epi-D homoestrones and their 3-methyl ethers. AB - An effective epimerization of the normal estrone 3-methyl and 3-benzyl ethers by using o-phenylenediamine and AcOH made the possibility for facile entry into the 13alpha-estrone series. Combination of this synthetic methodology with an isolation step carried out by means of the Girard-P reagent, the corresponding ethers of 13-epi-estrone were obtained in excellent yields. The 3-hydroxy and 3 methoxy D-homoestrone derivatives in both the normal and the 13alpha-estrone series were then synthesized and tested in vitro in a radioligand-binding assay. The estrogen receptor recognizes these compounds, but their relative binding affinities (RBAs) are lower than that of the reference compound 3,17beta estradiol. The progesterone receptor-binding affinities of the four D-homo derivatives were also tested showing low values for 13alpha-D-homoestrone and its 3-methyl ether. Pharmacologically, these 13alpha-D-homoestrone derivatives are estrogen receptor-selective molecules. PMID- 12628692 TI - Addition reactions at the 16(17) double bond of 3-methoxy-13alpha-estra 1,3,5(10),16-tetraene. AB - The epoxidation, the addition of hypobromous acid, and the hydroboration of 3 methoxy-13alpha-estra-1,3,5(10),16-tetraene 1 with diborane, catecholborane, and 9-BBN were investigated in order to determine the stereochemical outcome and to synthesize new 13alpha-estra-1,3,5(10)-trienes for biological and conformational investigations. It was shown that the sterically demanding reagent 9-BBN participated in a preferred beta attack (53% 16betaOH 10, 34% 17betaOH 8, 13% 16alphaOH 11). This stereochemical result is in agreement with that from another cis addition reaction, the recently described OsO4 dihydroxylation of 1 [Steroids 68 (2003) 113]. With smaller reagents such as B2H6, catecholborane, or magnesium monoperoxyphthalate, a diminished stereoselectivity was observed with only a slight excess of beta attack. The ionic trans addition of hypobromous acid gave two 17-bromo-16-alcohols with 16beta,17alpha (4, 76%) and 16alpha,17beta configuration (5, 24%) formed by trans cleavage of the 16,17alpha- and beta bromonium ion at position 16. The same regioselective and stereoselective course was found for the cleavage of the 16alpha,17alpha- and 16beta,17beta-epoxides (3 and 2) with hydrazoic acid (3-->16betaN3,17alphaOH 7, 2-->16alphaN3,17betaOH 6). The stereochemistry of the addition reactions to 1 can be explained in terms of a twist-boat conformation involving the C ring of compound 1. From a synthetic viewpoint the synthesis of the beta-epoxide 2 from the bromohydrin 4, the cleavage of this epoxide to 16alpha-substituted-17beta-hydroxy compounds, such as 6, and hydroboration/oxidation with 9-BBN to the hitherto unknown 16beta-hydroxy compound 10 are useful procedures. The bromohydrin 5 is the first 13alpha-steroid with a 17beta-bromo substituent. X-ray analysis revealed twist-boat and 16beta envelope conformations for rings C and D, respectively. PMID- 12628693 TI - Pheromones of the male sea lamprey, Petromyzon marinus L.: structural studies on a new compound, 3-keto allocholic acid, and 3-keto petromyzonol sulfate. AB - This study reports the results of chemical and chromatographic studies which establish the presence of 3-keto allocholic acid (3kACA) in water extracts from spermiating male sea lamprey, Petromyzon marinus. This is the second compound to be isolated and identified from these extracts. The first was 3-keto petromyzonol sulfate (3kPZS), which was shown to act as strong pheromonal attractant for ovulated females. Some new characterization data on 3kPZS (utilizing an only recently available synthetic preparation of the compound) is also included. The possibility that a mixture of 3kACA and 3kPZS might be a more potent pheromonal attractant than either compound alone is discussed. PMID- 12628695 TI - Mannose receptor determination by an ELISA-like method. AB - Mannose receptor determination may be a useful tool in research, because endocytosis via this animal lectin is involved in many functions of macrophage cells, in particular, the scavenger activity, the specific and unspecific defence against infective diseases, the recognition of neoplastic cells and the activation/differentiation process of the monocyte/macrophage and microglial population. To date, available tests required expensive equipment, the use of radioactive material or the availability of a specific antiserum. We describe an ELISA-like assay, based on biotinylated mannose-conjugated bovine serum albumin (BSA), which specifically binds to the cell mannose receptor. Biotin-labelled receptors can be quantified colorimetrically, utilising an avidin-alkaline phosphatase conjugate as the indicator enzyme. This new method is sensitive and reproducible, as well as simple and rapid, and can be performed with standard laboratory equipment. PMID- 12628694 TI - Diagnostic monitoring of urine by means of synchronous fluorescence spectrum. AB - A novel approach to clinical-biochemical analysis of urine is presented in this work. Urine composition is defined graphically as a record of synchronous fluorescence spectra (SFS). The graphical standard has been made from SFS of urine samples from healthy children. Simple comparison of a standard record with that of an analyzed urine sample will immediately reveal changes in its composition. Reproducibility of the graphical definition is very high and it maintains its characteristic shape during repeated measurements over a span of 2 years. It is possible to elaborate patients' own standard for those with chronic illness. It differs from a normal course but it is characteristic for a given patient and it enables the clinician to monitor changes or the outcome of therapy at regular medical examinations. Application of this method for monitoring of urine composition for selected cases is a new alternative with several advantages. Analysis without any added reagents very quickly detects some illnesses near onset when they may be clinically asymptomatic and classical screening methods show negative results. Computerization of spectral measuring and filing the results enables to give a likely diagnosis or a deviation from standard. This method can also serve a doctor-clinician either to confirm or to exclude a concrete diagnosis. PMID- 12628696 TI - Centrifugal and chromatographic analyses of tryptophan and tyrosine uptake by red blood cells and GLUT1 proteoliposomes with permeability estimates and observations on dihydrocytochalasin B. AB - We analyzed transport into liposomes and proteoliposomes, separated the free and internalized radioactively labeled substrates by size-exclusion chromatography (SEC) and observed a net influx owing to nonfacilitated diffusion across the lipid bilayers during the separation. The permeabilities (10(-9) cm/s) of glucose transporter (GLUT1) proteoliposomes were estimated to be 4.6, 1.0, 1.4 and 2.1 for D-glucose, L-glucose, L-Tyr and L-Trp, respectively; 15, 3.3, 5.1 and 2.1 times higher than the corresponding permeabilities of liposomes. These values indicated that GLUT1 did not transport Tyr or Trp, or transported Tyr, and only Tyr, slowly. This interpretation was supported by further analyses. Dihydrocytochalasin B inhibited the transport of Tyr and, partially, Trp into human red blood cells (centrifugal analyses). It did not inhibit Tyr and Trp influx into GLUT1 proteoliposomes, but partitioned strongly into the bilayers and seemed to make them fragile. The GLUT1 inhibitor cytochalasin B and the GLUT1 substrate 2-deoxy-D-glucose did not inhibit Tyr transport into the cells. Upon immobilized biomembrane affinity chromatography, Trp decreased the cytochalasin B retardation by GLUT1 only at levels far above the physiological Trp concentration. Ethanol (commonly added to aqueous solutions for enhancing a compound's solubility) halved the retardation at 4% (v/v) concentration. Drastic modification of the SEC method is required to allow permeability measurements with nonlabeled and highly permeable substrates. PMID- 12628698 TI - Efficient and accurate experimental design for enzyme kinetics: Bayesian studies reveal a systematic approach. AB - In areas such as drug development, clinical diagnosis and biotechnology research, acquiring details about the kinetic parameters of enzymes is crucial. The correct design of an experiment is critical to collecting data suitable for analysis, modelling and deriving the correct information. As classical design methods are not targeted to the more complex kinetics being frequently studied, attention is needed to estimate parameters of such models with low variance. We demonstrate that a Bayesian approach (the use of prior knowledge) can produce major gains quantifiable in terms of information, productivity and accuracy of each experiment. Developing the use of Bayesian Utility functions, we have used a systematic method to identify the optimum experimental designs for a number of kinetic model data sets. This has enabled the identification of trends between kinetic model types, sets of design rules and the key conclusion that such designs should be based on some prior knowledge of K(M) and/or the kinetic model. We suggest an optimal and iterative method for selecting features of the design such as the substrate range, number of measurements and choice of intermediate points. The final design collects data suitable for accurate modelling and analysis and minimises the error in the parameters estimated. PMID- 12628697 TI - FTIR microspectroscopy of malignant fibroblasts transformed by mouse sarcoma virus. AB - Fourier transform infrared microspectroscopy (FTIR-MSP), which is based on the characteristic molecular vibrational spectra of cells, was used to investigate spectral differences between normal primary rabbit bone marrow (BM) cells and bone marrow cells transformed (BMT) by murine sarcoma virus (MuSV). Primary cells, rather than cell lines, were used for this research because primary cells are similar to normal tissue cells in most of their characteristics. Our results showed dramatic changes in absorbance between the control cells and MuSV124 transformed cells. Various biological markers, such as the phosphate level and the RNA/DNA obtained, based on the analysis of the FTIR-MSP spectra, also displayed significant differences between the control and transformed cells. Preliminary results suggested that the cluster analysis performed on the FTIR-MSP spectra yielded 100% accuracy in classifying both types of cells. PMID- 12628699 TI - Patch-clamp recordings in isolated sponge cells (Axinella polypoides). AB - Sponges are the most ancient known metazoans. Their cells are specialised but not organised into tissues or organs. Recordings of action potential-like propagating electrical impulses suggested that electrical signalling may occur between sponge cells, but the characterization of ionic channels in these cells is still at the beginning. Actually, sponge cell surfaces are covered by a complex glycocalyx and long-chain fatty acids are present in the lipid core of their membranes. In these experimental conditions, a low percentage of tight seals (3%) was obtained applying the patch-clamp technique to cells isolated from the Mediterranean Demospongia Axinella polypoides. This paper shows in detail how difficulties can be overcome making use of trivalent cations in the extracellular solution and how electrophysiological measurements can be performed on sponge cell membranes. A potassium selective conductance is shown as an example. We suggest that the presented methodology could also be applied to other cell types. PMID- 12628700 TI - Interaction of Al(III) with the peptides AspAsp and AspAspAsp. AB - The interactions of Al(III) with the dipeptide AspAsp and the tripeptide AspAspAsp in aqueous solutions were studied by pH-potentiometry and multinuclear 1H- and 13C- nuclear magnetic resonance (NMR) spectroscopy. Their numerous negatively charged COO(-) functions allow these ligands to bind Al(III) even in weakly acidic solutions. Various mononuclear 1:1 complexes are formed in different protonation states. 13C-NMR spectroscopy unambiguously proved participation of the COO(-) functions in a monodentate or chelating mode in Al(III) binding, however, the terminal-NH(2) group seems to be excluded from the coordination. Depending on the metal ion to ligand ratio precipitation occurs at pH approximately 5 to 6. This indicates that the COO(-) groups at the low level of preorganization in such small peptides are not sufficient to keep the Al(III) ion in solution and to prevent the precipitation of Al(OH)(3) at physiological pH. To achieve this, a more specific arrangement of the side-chain donors seems necessary. PMID- 12628701 TI - Borate in mummification salts and bones from Pharaonic Egypt. AB - Mummification processes in Pharaonic Egypt were successful using sodium salts. Quite frequently sodium concentrations in mummified bones ranged from 300 to 4000 micromol/g. In the search for an effective inorganic conservation compound our choice fell on boric acid. The possible presence of borate in mummification salts used in Pharaonic Egypt was of special interest both historically and biochemically. In two salt samples, one from the embalming material of Tutankhamen (18th dynasty, 1336-1327 BC) and the second from Deir el-Bahari (25th dynasty, 700-600 BC) borate was found, amounting to 2.1+/-0.2 and 3.9+/-0.1 micromol/g, respectively. In five of the examined bone fragments from the Junker excavation at Giza (Old Kingdom) similar borate concentrations i.e., 1.2 micromol borate/g bone were seen. It must be emphasized that the usual borate content of contemporary autopsy is far below the detection limit. The elevated borate content in both mummification salt and ancient bone samples support the suggestion that borate-containing salt had been used. There is a striking correlation of both borate concentration and alkaline phosphatase activity. When both sodium salts and borate were essentially absent no activity at all was detectable. With increasing borate concentrations the enzyme activity rises significantly. Attributable to the distinct biochemistry of the tetrahydroxyborate anion it was of interest whether or not borate may stabilize alkaline phosphatase, an important and richly abundant bone enzyme. This enzyme was chosen, as it is known to survive more than 4000 years of mummification. In the presence of borate oligomeric species of this zinc-magnesium-glycoprotein at 400,000 Da became detectable. Attributable to this borate-dependent stabilization of the enzyme molecule a significant temperature resistant increase of the enzymic activity was measured in the presence of up to 2.5 mM borate. PMID- 12628702 TI - Effects of analogues of inorganic phosphate and sodium ion on mineralization of matrix vesicles isolated from growth plate cartilage of normal rapidly growing chickens. AB - The mechanism of matrix vesicle (MV) mineralization was studied using MVs isolated from normal growth plate tissue, as well as several putative intermediates in the MV mineralization pathway--amorphous calcium phosphate (ACP), calcium phosphate phosphatidylserine complex (CPLX) and hydroxyapatite (HAP). Radionuclide uptake and increase in turbidity were used to monitor mineral formation during incubation in synthetic cartilage lymph (SCL). Inhibitors of phosphate (Pi) metabolism, as well as replacing Na(+) with various cations, were used to study MV Pi transport, which had been thought to be Na(+)-dependent. MVs induced rapid mineralization approximately 3 h after addition to SCL; CPLX and HAP caused almost immediate induction; ACP required approximately 1 h. Phosphonoformate (PFA), a Pi analog, potently delayed the onset and reduced the rate of mineral formation of MV and the intermediates with IC(50)'s of 3-6 microM and approximately 10 microM, respectively. PFA:Pi molar ratios required to reduce the rate of rapid mineralization by 50% were approximately 1:30 for ACP, approximately 1:20 for HAP, approximately 1:3.3 for CPLX, and approximately 1:2.0 for MVs. MV mineralization was not found to be strictly Na(+)-dependent: substitution of Li(+) or K(+) for Na(+) had minimal effect; while N-methyl D glucamine (NMG(+)) was totally inhibitory, choline(+) was clearly stimulatory. Na(+) substitutions had minimal effect on HAP- and CPLX-seeded mineral formation. However with ACP, NMG(+) totally blocked and choline(+) stimulated, just as they did MV mineralization. Thus, kinetic analyses indicate that ACP is a key intermediate, nevertheless, formation of CPLX appears to be the rate-limiting factor in MV mineralization. PMID- 12628703 TI - Activity, stability and conformational flexibility of seed coat soybean peroxidase. AB - Seed coat soybean peroxidase (SBP) belongs to class III of the plant peroxidase superfamily that includes the classical peroxidase, namely horseradish peroxidase (HRP). We have measured the catalytic activity (k(cat)) and catalytic efficiency (k(cat)/K(M)) of SBP and that of HRP-C for the oxidation of ABTS [2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonate)] by hydrogen peroxide at 25 degrees C. We observed that the k(cat) and k(cat)/K(M) values for SBP are much higher than those for HRP-C at all pH values, rendering SBP a more potent peroxidase. This is attributed to the relatively more solvent exposed delta-meso heme edge in SBP. We observed that the maximum catalytic activity and conformational stability of SBP is at pH approximately 5.5. A pH maximum of 5.0 for the catalytic activity of SBP has recently been reported. Estimation of secondary structural elements at various pH values indicated that there is a maximal reduction of beta-strands and beta-turns at pH 5.5 causing the heme to be further exposed to the solvent and increasing the overall conformational flexibility of the protein. PMID- 12628704 TI - Purification and characterization of a new cationic peroxidase from fresh flowers of Cynara scolymus L. AB - A basic heme peroxidase isoenzyme (AKPC) has been purified to homogeneity from artichoke flowers (Cynara scolymus L.). The enzyme was shown to be a monomeric glycoprotein, M(r)=42300+/-1000, (mean+/-S.D.) with an isoelectric point >9. The native enzyme exhibits a typical peroxidase ultraviolet-visible spectrum with a Soret peak at 404 nm (epsilon=137,000+/-3000 M(-1) cm(-1)) and a Reinheitzahl (Rz) value (A(404nm)/A(280nm)) of 3.8+/-0.2. The ultraviolet-visible absorption spectra of compounds I, II and III were typical of class III plant peroxidases but unlike horseradish peroxidase isoenzyme C, compound I was unstable. Resonance Raman and UV-Vis spectra of the ferric form show that between pH 5.0 and 7.0 the protein is mainly 6 coordinate high spin with a water molecule as the sixth ligand. The substrate-specificity of AKPC is characteristic of class III (guaiacol-type) peroxidases with chlorogenic and caffeic acids, that are abundant in artichoke flowers, as particularly good substrates at pH 4.5. Ferric AKPC reacts with hydrogen peroxide to yield compound I with a second-order rate constant (k(+1)) of 7.4 x 10(5) M(-1) s(-1) which is significantly slower than that reported for most other class III peroxidases. The reaction of ferric and ferrous AKPC with nitric oxide showed a potential use of this enzyme for quantitative spectrophotometric determination of NO and as a component of novel NO sensitive electrodes. PMID- 12628705 TI - Synthesis, characterization and in vitro cytotoxic, mutagenic and antimicrobial activity of platinum(II) complexes with substituted benzimidazole ligands. AB - In this study, six Pt(II) complexes bearing 5(6)-H or -CH(3)-2-phenyl or -(2' pyridyl) or -mercaptomethylbenzimidazole ligands as 'carrier groups' were synthesized and characterized by elemental analysis, IR and (1)H-NMR spectra and evaluated for their preliminary in vitro cytotoxic activity to the human RD Rhabdomyosarcoma cell line and mutagenic properties in Salmonella typhimurium strains TA 98 and TA 100 in the absence of the S9 rat liver fraction. The preliminary test results showed that the complexes had slightly greater cytotoxic activity on the RD cell line at 1 microM concentration than cisplatin. Among the compounds tested for their mutagenicity, Pt(II) complexes of 2-(2'-pyridyl)- and 5(6)-methyl-2-(2'-pyridyl)benzimidazoles were found to be mutagenic. A comparative study of the MIC (minimum inhibitory concentration) values indicated that, in general, there were no differences between the poor antimicrobial activity values of the ligands and their Pt(II) complexes with respect to the tested microorganisms. These results suggest that the synthesized Pt(II) complexes should be considered for further antitumor activity studies. PMID- 12628706 TI - DNA binding of iron(II) complexes with 1,10-phenanthroline and 4,7-diphenyl-1,10 phenanthroline: salt effect, ligand substituent effect, base pair specificity and binding strength. AB - The DNA binding of iron(II) mixed-ligand complexes containing 1,10 phenanthroline(phen) and 4,7-diphenyl-1,10-phenanthroline(dip), [Fe(phen)(3)](2+), [Fe(phen)(2)(dip)](2+) and [Fe(phen)(dip)(2)](2+) has been characterized by spectrophotometric titration and melting temperature measurements. The salt concentration dependence of the binding constant has allowed us to dissect the DNA-binding constant and free energy change of each iron(II) complex into the nonelectrostatic and polyelectrolyte contributions. A comparison of the nonelectrostatic components in the binding free energy changes among iron(II) complexes has made it possible to rigorously evaluate the contribution of the ligand substituents to the DNA-binding event. The peripheral substitution of phen by two phenyl groups increases the nonelectrostatic binding constant of the iron(II) complex more than 20 times, which is equivalent to approximately 7.5 kJ mol(-1) of more favorable contribution to the DNA binding. In general, the iron(II) complexes studied have higher affinity towards the more facile A-T sequence than the G-C sequence. This preferential binding may be attributed to the steric effect induced by the ancillary part of the ligands in the course of DNA binding. The binding of disubstituted iron(II) complex to DNA is quite strong as reflected in the modest increase in the denaturation temperature (T(m)) of double helical DNA upon the interaction with the iron(II) complex. PMID- 12628707 TI - Synthesis and DNA binding properties of a cationic 2,2':6',2"-terpyridine cobalt(II) complex containing an oligopeptide. AB - The first example of DNA metallointercalator containing an oligopeptide moiety is presented. The ternary cobalt(II) complex [(DOTA)Co(II)(TPY)](ClO(4))(2) (DOTA=1,4,7,10-tetraaza-cyclododecane-2,9-dione, TPY=2, 2':6',2"-terpyridine) was found to be efficient in binding ct-DNA. The binding constant was determined by spectrophotometric titration. In vitro antitumor studies shows that this complex has significant antitumor activity. The single-crystal of a six coordinated oligopeptide cobalt(II) complex [Co(II)(DOTA)(2)](ClO(4))(2) is also reported. The stabilities and species distributions of Co(II)-DOTA (1:1) and DOTA-Co(II) TPY (1:1:1) systems were investigated by potentiometry titration. PMID- 12628708 TI - Synthesis and antitumor activity of cyclopentadienyltitanium substituted polyoxotungstate [CoW11O39(CpTi)]7- (Cp=eta5-C5H5). AB - A novel polyoxotungstate [CoW(11)O(39)(CpTi)](7-) (Cp=eta(5)-C(5)H(5)) has been prepared. This complex exhibits the highest antitumor activity in vitro among the cyclopentadienyltitanium substituted polyoxometalates investigated and has a remarkable inhibitory action on three types of human cancer cells, SSMC-7721, HL 60 and HLC, in vivo. PMID- 12628709 TI - Cadmium and vanadate oligomers effects on methaemoglobin reductase activity from Lusitanian toadfish: in vivo and in vitro studies. AB - Cadmium and two vanadate solutions as 'metavanadate' (containing ortho and metavanadate species) and 'decavanadate' (containing decameric species) (5 mM) were injected intraperitoneously in Halobatrachus didactylus (Lusitanian toadfish), in order to evaluate the effects of cadmium and oligomeric vanadate species on methaemoglobin reductase activity from fish red blood cells. Following short-term exposure (1 and 7 days), different changes were observed on enzyme activity. After 7 days of exposure, 'metavanadate' increased methaemoglobin reductase activity by 67% (P < 0.05), whereas, minor effects were observed on enzymatic activity upon cadmium and 'decavanadate' administration. However, in vitro studies indicate that decameric vanadate, in concentrations as low as 50 microM, besides strongly inhibiting methaemoglobin reductase activity, promotes haemoglobin oxidation to methaemoglobin. Although decameric vanadate species showed to be unstable in the different media used in this work, the rate of decameric vanadate deoligomerization is in general slow enough, making it possible to study its effects. It is concluded that the increase in H. didactylus methaemoglobin reductase activity is more pronounced upon exposition to 'metavanadate' than to cadmium and decameric species. Moreover, only decameric vanadate species promoted haemoglobin oxidation, suggesting that vanadate speciation is important to evaluate in vivo and in vitro effects on methaemoglobin reductase activity. PMID- 12628710 TI - Thermodynamic, kinetic and structural studies on the mixed ligand complexes of palladium(II) with tridentate and monodentate ligands. AB - Stability constants of the complexes formed in the reaction of [Pd(bpma)](2+) [bpma=bis(pyridin-2-ylmethyl)amine] with monodentate nitrogen and thioether ligands including uridine, MeUH, cytidine, MeC, EtGH, AcHis, AcHm, AcLys and AcMet were determined by potentiometric method. The coordination chemistry of [Pd(bpma)](2+) shows a significant similarity to that of [Pd(terpy)](2+), but it is different from [Pd(dien)](2+). The formation of hydroxo and dinuclear complexes is especially enhanced in the case of [Pd(bpma)](2+) and [Pd(terpy)](2+), but the affinity of palladium(II) ions for the coordination of thioether residues is reduced in the presence of pyridine nitrogen atoms. Stopped flow kinetic measurements reveal that the substitution reactions of the thioether ligand AcMet are much faster than those of the N-donor cytidine. The presence of the two pyridyl residues significantly enhances the kinetic reactivity of [Pd(bpma)](2+) as compared to that of [Pd(dien)](2+). The Pd-S(thioether) bonded species can be important intermediates in multicomponent systems, but the equilibrium state is characterised by the formation of Pd-N bonded species. The complex [Pd(bpma)NO(3)]NO(3) has been prepared in solid state and its structure was elucidated by single crystal X-ray diffraction method. PMID- 12628711 TI - Transplantation of cells for cardiac repair. AB - The inability of adult cardiomyocytes to divide to a significant extent and regenerate the myocardium after injury leads to permanent deficits in the number of functional cells, which can contribute to the development and progression of heart failure. The transplantation of skeletal myoblasts or stem cells or cardiomyocytes derived from them into the injured myocardium is a novel and promising approach in the treatment of cardiac disease and the restoration of myocardial function. In this article, skeletal myoblasts and embryonic and bone marrow stem cells are discussed in the context of their potential therapeutic use in cardiac failure. The state of the art in both laboratory and clinic is presented. We discuss current and intrinsic limitations of cardiac cellular transplantation and suggest directions for future research. PMID- 12628712 TI - Influence of concurrent renal dysfunction on outcomes of patients with acute coronary syndromes and implications of the use of glycoprotein IIb/IIIa inhibitors. AB - OBJECTIVES: The purpose of this study was to examine the in-hospital outcome and influence of glycoprotein (GP) IIb/IIIa antagonists on patients with acute coronary syndromes (ACS) across a range of renal function. BACKGROUND: Recent studies demonstrate increasing cardiovascular risk with progressive renal dysfunction. Previous studies investigating GP IIb/IIIa antagonist use have excluded patients with renal dysfunction. METHODS: Patients presenting with ACS between January 1999 and May 2000 were identified, and data on demographics, in hospital management, and clinical events were collected using standardized definitions. Patients were stratified according to renal function assessed by calculated creatinine clearance (CrCl) at presentation. Primary outcome measures included in-hospital mortality and major bleeding events. RESULTS: Renal insufficiency was present in 312 of 889 patients. There were 40 in-hospital deaths. In non-dialysis-dependent patients, as CrCl worsened, there was a decline in utilization of routine diagnostics and therapeutics, an increase in in hospital mortality (p = 0.002), and an increase in major bleeding (p = 0.03). Although the use of GP IIb/IIIa antagonists was associated with an increase in major bleeding (p < 0.001), there was a protective effect on in-hospital mortality (p = 0.04) after controlling for CrCl. CONCLUSIONS: Renal dysfunction is present in a substantial proportion of patients with ACS and is associated with increased in-hospital death. Although GP IIb/IIIa antagonist use in patients with ACS and renal insufficiency resulted in increased bleeding events, its administration was associated with a decreased risk of in-hospital mortality. These preliminary findings need to be confirmed in future randomized clinical trials. PMID- 12628713 TI - Why is chronic kidney disease the "spoiler" for cardiovascular outcomes? PMID- 12628714 TI - Angiotensin-converting enzyme gene polymorphism interacts with left ventricular ejection fraction and brain natriuretic peptide levels to predict mortality after myocardial infarction. AB - OBJECTIVES: The goal of this study was the exploration of the associations between the angiotensin-converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism and post-myocardial infarction (MI) outcomes, especially any interaction with the accepted clinical prognostic markers brain natriuretic peptide (BNP) and left ventricular ejection fraction (LVEF). BACKGROUND: The ACE gene I/D polymorphism has been implicated in the development of MI, hypertension, and left ventricular hypertrophy. We examined the association of ACE I/D and prognosis after acute MI. METHODS: Patients incurring acute MI were genotyped for the ACE I/D polymorphism. Clinical data included assays of neurohormones, radionuclide ventriculography, and mortality over a mean 2.6 years of follow-up. RESULTS: Patients (n = 978) had a mean age of 62.1 years, and 78% were male. Overall genotype frequencies were II 23.2%, ID 49.5%, and DD 27.3%. Chi-square analysis revealed an association between the ACE D allele and death after MI (88 of 103 who died were DD or ID; p < 0.05), with an odds ratio for mortality of 8.03 (95% confidence interval, 2.16 to 29.88). Patients with the DD genotype had higher (p < 0.05) plasma BNP, N-terminal BNP (N-BNP), and endothelin-1 levels within 96 h after MI than grouped ID/II patients. Multivariate analysis indicated ACE genotype, age, and previous MI were independent predictors of death (p < 0.05). Patients with an ACE D allele in combination with either a lower than median LVEF or greater than median BNP had a higher mortality (p < 0.001 and p < 0.025, respectively) than the risk associated with the D allele itself. CONCLUSIONS: Angiotensin-converting enzyme genotyping may provide additional prognostic information in patients after MI in combination with the proven utility of LVEF, plasma BNP, and N-BNP measurements. PMID- 12628715 TI - Assessment of clinical features in transient left ventricular apical ballooning. AB - OBJECTIVES: We sought to assess the clinical features of transient left ventricular (LV) apical ballooning. BACKGROUND: Although several cases regarding transient LV apical ballooning have been reported, the etiology remains unknown. METHODS: We investigated 17 patients (14 women, median age 74 years old with a range of 54 to 91 years old) who fulfilled the following criteria: 1) transient LV apical ballooning; 2) ST-T segment change in several leads in electrocardiogram; and 3) no history of old myocardial infarction, valvular heart disease, subarachnoid hemorrhage, or pheochromocytoma. RESULTS: Emotional and physical stress were observed in 16 patients (94%). Technetium-99m tetrofosmin tomographic imaging revealed decreased uptake at the apex of the left ventricle in 11 patients (85%) that later returned to uniform. No significant stenosis or angiographical slow flow in epicardial coronary arteries was observed (n = 9). Provocative focal vasospasm was induced in only one patient (14%) (n = 7). Moreover, no significant abnormality in the coronary microcirculation was detected by Doppler guidewire (n = 3) or contrast echocardiography (n = 1). No patients showed a rise in viral antibody titers. Biopsy specimens revealed interstitial fibrosis in six patients (100%) and slight cell infiltration in three others (50%) (n = 6). CONCLUSIONS: These findings suggested that neither abnormalities in the coronary circulation nor acute myocarditis was related to the etiology. Although neurogenic stunned myocardium induced by emotional or physical stress was suggested as the etiology, further investigations are necessary. PMID- 12628716 TI - Myocardial perfusion and fatty acid metabolism in patients with tako-tsubo-like left ventricular dysfunction. AB - OBJECTIVES: We sought to assess myocardial perfusion and metabolism in patients with peculiar transient asynergy, which consisted of basal normokinesia and apical akinesia of the left ventricle (LV) at the same time. BACKGROUND: This asynergy has been widely called "tako-tsubo-like LV dysfunction" in Japan, but little is known about its pathophysiology. METHODS: We performed rest tallium-201 ((201)Tl) and iodine-123-beta-methyl-p-iodophenyl penta-decanoic acid ((123)I BMIPP) dual-isotope myocardial single-photon emission computed tomography (SPECT) in 14 patients with tako-tsubo-like LV dysfunction. The LV was divided into 17 segments, and each segment was graded with scores between normal uptake (0) and defect (4). We also measured the Thrombolysis in Myocardial Infarction trial (TIMI) frame count in 28 patients and 20 control subjects. RESULTS: Early SPECT (5 +/- 3 days) revealed that the total defect score value with BMIPP was significantly higher than reduced uptake with (201)Tl (p < 0.01). Reduced uptake of BMIPP was observed in parallel with an apical akinetic region and usually involved uptake of (201)Tl. This discrepancy improved gradually during the follow up period (29 +/- 6 days) (p = 0.36). Compared with control subjects, patients had a TIMI frame count that was significantly higher in all coronary arteries immediately after onset. This higher TIMI frame count decreased but was sustained even after resolution of tako-tsubo-like LV dysfunction. CONCLUSIONS: Our data suggest that myocardial fatty acid metabolism is more severely impaired than myocardial perfusion, in parallel with an apical akinetic region during the early phase, and that impaired multivessel coronary microcirculation is involved, at least in part, in tako-tsubo-like LV dysfunction. PMID- 12628717 TI - Effect of percutaneous coronary interventions for in-stent restenosis in degenerated saphenous vein grafts without distal embolic protection. AB - OBJECTIVES: This study was designed to investigate the impact of percutaneous coronary interventions (PCIs) in degenerated saphenous vein grafts (SVGs) without distal embolic protection. BACKGROUND: Distal embolic protection devices have been shown to reduce the incidence of no reflow/slow flow during PCI of de novo lesions in degenerated SVGs. It is unclear whether PCI of in-stent restenosis (ISR) lesions in degenerated SVGs is associated with no reflow/slow flow and whether distal embolic protection is beneficial in these cases as well. METHODS: We studied 54 consecutive patients with treated ISR lesions in degenerated SVGs who underwent PCI without distal embolic protection in a single center. Procedural and in-hospital outcomes were examined. The average age was 71 +/- 8 years; 32% of the patients had diabetes. The mean lesion length was 13 +/- 6 mm and the procedural success rate was 98% (53/54). Cutting balloon angioplasty was used in 46% (25/54) of cases, and a new stent was inserted in 46% (25/54) of patients. Gamma brachytherapy was performed in 19% (10/54) of patients. During the procedure there were no episodes of no reflow/slow flow, and there were no patients with in-hospital Q-wave or non-Q-wave myocardial infarction. There was one in-hospital noncardiac death. CONCLUSIONS: In this consecutive series of patients with ISR of degenerated SVGs undergoing PCI without distal protection, there were no episodes of slow flow/no reflow and no procedure-related myocardial infarctions. It appears that distal embolic protection may not be necessary during PCI of ISR lesions in degenerated SVGs. PMID- 12628718 TI - Increased myocardial apoptosis in patients with unfavorable left ventricular remodeling and early symptomatic post-infarction heart failure. AB - OBJECTIVES: The purpose of this study was to evaluate a potential correlation between apoptotic rate (AR), post-infarction left ventricular (LV) remodeling, and clinical characteristics in subjects who died late (>or=10 days) after an acute myocardial infarction (AMI) with evidence of persistent occlusion of the infarct-related artery at autopsy. BACKGROUND: Apoptosis contributes to myocardiocyte loss in cardiac disease and may have a pathophysiologic role in post-infarction LV remodeling. METHODS: The AR was calculated at the site of infarction and in remote unaffected LV regions, using co-localization of in situ end labeling for deoxyribonucleic acid fragmentation and immunohistochemistry for caspase-3, in 14 subjects who died within two months after AMI. Correlation between AR and clinical characteristics such as age, site of AMI, transmural extension, multivessel coronary disease, and signs and/or symptoms of heart failure (HF), at the time of initial hospitalization for AMI or subsequently before death, was assessed using non-parametric statistical tests. Parameters of LV remodeling including diameters, free wall thickness, diameter-to-wall thickness ratio, and mass were measured at gross examination at autopsy. Values are expressed as median (interquartile range). RESULTS: Among clinical variables, early symptomatic post-infarction HF (9 cases, 64%) was associated with nearly fourfold increased AR at the site of infarction (26.2% [24.5% to 28.8%] vs. 6.4% [1.9% to 13.3%], p = 0.001). Moreover, AR both at the site of infarction and in unaffected regions was significantly correlated with parameters of progressive LV remodeling (p < 0.05). CONCLUSIONS: Our data show that in patients dying >or=10 days after AMI, myocardial apoptosis is strongly associated with and may be a major determinant of unfavorable LV remodeling and early symptomatic post infarction HF. PMID- 12628719 TI - Myocyte apoptosis: programming ventricular remodeling. PMID- 12628720 TI - Acute effects of cardiac resynchronization therapy on functional mitral regurgitation in advanced systolic heart failure. AB - OBJECTIVES: We studied the acute effects of cardiac resynchronization therapy (CRT) on functional mitral regurgitation in heart failure (HF) patients with left bundle branch block (LBBB). BACKGROUND: Both an decrease [corrected] in left ventricular (LV) closing force and mitral valve tethering have been implicated as mechanisms for functional mitral regurgitation (FMR) in dilated hearts. We hypothesized that an increase in LV closing force achieved by CRT could act to reduce FMR. METHODS: Twenty-four HF patients with LBBB and FMR were studied after implantation of a biventricular CRT system. Acute changes in FMR severity between intrinsic conduction (OFF) and CRT were quantified according to the proximal isovelocity surface area method by measuring the effective regurgitant orifice area (EROA). Results were compared with the changes in estimated maximal rate of left ventricular systolic pressure rise (LV+dP/dt(max)) and transmitral pressure gradients (TMP), both measured by Doppler echocardiography. RESULTS: Cardiac resynchronization therapy was associated with a significant reduction in FMR severity. Effective regurgitant orifice area decreased from 25 +/- 19 mm(2) (OFF) to 13 +/- 8 mm(2) (CRT). The change in EROA was directly related to the increase in LV+dP/dt(max) (r = -0.83, p < 0.0001). Compared with OFF, TMP increased more rapidly during CRT, and a higher maximal TMP was observed (OFF 73 +/- 24 mm Hg vs. CRT 85 +/- 26 mm Hg, p < 0.01). CONCLUSIONS: Functional mitral regurgitation is reduced by CRT in patients with HF and LBBB. This effect is directly related to the increased closing force (LV+dP/dt(max)). The results support the hypothesis that an increase in TMP, mediated by a rise in LV+dP/dt(max) due to more coordinated LV contraction, may facilitate effective mitral valve closure. PMID- 12628721 TI - Natural history of dilated cardiomyopathy due to lamin A/C gene mutations. AB - OBJECTIVES: We examined the prevalence, genotype-phenotype correlation, and natural history of lamin A/C gene (LMNA) mutations in subjects with dilated cardiomyopathy (DCM). BACKGROUND: Mutations in LMNA have been found in patients with DCM with familial conduction defects and muscular dystrophy, but the clinical spectrum, prognosis, and clinical relevance of laminopathies in DCM are unknown. BACKGROUND: A cohort of 49 nuclear families, 40 with familial DCM and 9 with sporadic DCM (269 subjects, 105 affected), was screened for mutations in LMNA using denaturing high-performance liquid chromatography and sequence analysis. Bivariate analysis of clinical predictors of LMNA mutation carrier status and Kaplan-Meier survival analysis were performed. RESULTS: Mutations in LMNA were detected in four families (8%), three with familial (R89L, 959delT, R377H) and one with sporadic DCM (S573L). There was significant phenotypic variability, but the presence of skeletal muscle involvement (p < 0.001), supraventricular arrhythmia (p = 0.003), conduction defects (p = 0.01), and "mildly" DCM (p = 0.006) were predictors of LMNA mutations. The LMNA mutation carriers had a significantly poorer cumulative survival compared with non-carrier DCM patients: event-free survival at the age of 45 years was 31% versus 75% in non-carriers. CONCLUSIONS: Mutations in LMNA cause a severe and progressive DCM in a relevant proportion of patients. Mutation screening should be considered in patients with DCM, in particular when clinical predictors of LMNA mutation are present, regardless of family history. PMID- 12628722 TI - A novel missense mutation in the myosin binding protein-C gene is responsible for hypertrophic cardiomyopathy with left ventricular dysfunction and dilation in elderly patients. AB - OBJECTIVES: We studied the clinical features of hypertrophic cardiomyopathy (HCM) caused by a novel mutation in the myosin binding protein-C (MyBP-C) gene in patients and family members of Japanese descent. BACKGROUND: Previous reports have demonstrated that the clinical features of HCM associated with mutations in the MyBP-C gene include late onset and a favorable clinical course. Recently, some mutations in genes encoding sarcomeric proteins have been reported to be a cause of dilated cardiomyopathy (DCM), as well as HCM. However, mutations of the MyBP-C gene have not been reported as a cause of DCM up to now. METHODS: We analyzed MyBP-C gene mutations in 250 unrelated probands with HCM and in 90 with DCM. We used electrocardiography (ECG) and echocardiography to determine clinical phenotypes. RESULTS: We identified 17 individuals in 8 families (7 HCM, 1 DCM) with an Arg820Gln mutation in the MyBP-C gene. Overall, 2 (40%) of 5 carriers age >70 years displayed "burnt-out" phase HCM, and one of them had been diagnosed as having DCM before genetic identification. The disease penetrance in subjects age >50 years was 70% by echocardiography and 100% by ECG, and that in those age <50 years was 40% and 50%, respectively. CONCLUSIONS: Elderly patients with Arg820Gln mutation may show "burnt-out" phase HCM, and patients with this mutation may be included among those diagnosed as having DCM. Screening of patients with DCM, as well as HCM, for this mutation is of significant importance because patients with this mutation may be diagnosed clinically as having DCM. PMID- 12628724 TI - Syncope: is a diagnosis a diagnosis? PMID- 12628723 TI - Selective use of diagnostic tests inpatients with syncope of unknown cause. AB - OBJECTIVE: We sought to prospectively assess the diagnostic yielding of a protocol in which electrophysiologic studies (EPS), tilt-table tests (TTTs), and loop recorder implantation are selectively used. BACKGROUND: The optimal strategy in the diagnosis of patients with syncope of unknown cause has not been defined. METHODS: A total of 184 consecutive patients with syncope of unknown cause were classified into two groups. Group A consisted of 72 patients fulfilling any of the following criteria: 1) presence of structural heart disease or family history of sudden death; 2) abnormal electrocardiogram; 3) significant non-symptomatic arrhythmia on Holter monitoring; and 4) paroxysmal palpitations immediately before or after syncope. These patients initially underwent an EPS and, if this study was negative, TTT. In the remaining 112 patients (group B), TTT was performed. RESULTS: The EPS was positive in 32 patients (44%) in group A. The TTT was positive in 80 patients (71%) in group B. An additional patient had carotid sinus hypersensitivity. In patients of group A with a negative EPS, the TTT was positive in 23 (57%). A loop recorder was implanted in 15 patients from group A with negative conventional testing, and diagnostic activation was obtained in seven patients. Overall, a positive diagnosis was achieved in 143 patients (78%). CONCLUSIONS: In patients with syncope of unknown cause, selective use of EPS or TTT leads to a positive diagnosis in >70% of the cases. An implantable loop recorder can be useful in non-diagnosed cases. PMID- 12628725 TI - Development and validation of a simple risk score to predict the need for permanent pacing after cardiac valve surgery. AB - OBJECTIVES: The study objective was to develop and validate a simple risk score to predict postoperative permanent pacing (PPM) after valve surgery. BACKGROUND: Our ability to identify patients preoperatively that will require PPM is poor. A simple preoperative risk score to predict PPM after valve surgery could assist both clinical practice and research. METHODS: All valve surgery patients at our institution from 1992 to 2002 were included (n = 4,694). Two-thirds of the patients were randomly selected to form a risk score prediction group (PG), and the score was then applied to the remaining patients (validation group [VG]). RESULTS: Preoperative right bundle branch block (odds ratio [OR], 3.6; 95% confidence interval [CI], 2.3 to 5.7) and multivalve surgery that included the tricuspid valve (OR, 3.7; 95% CI, 2.3 to 6.1) were the strongest independent predictors of PPM, while multivalve surgery that did not include the tricuspid valve (OR, 2.1; 95% CI, 1.3 to 3.3), preoperative left bundle branch block (OR, 2.0; 95% CI, 1.3 to 2.9), preoperative PR interval >200 ms (OR, 1.9; 95% CI, 1.3 to 3.0), prior valve surgery (OR, 1.8, 95% CI, 1.2 to 2.7), and age >70 years (OR, 1.4; 95% CI, 1.04 to 2.0) also predicted PPM. A risk score from 0 to 6 identified patients in the VG with incidences of PPM of 1.9%, 5.2%, 8.7%, 11.5%, 21%, 36%, and 50%, respectively. CONCLUSIONS: A simple risk score incorporating preoperative conduction, age, prior valve surgery, and surgery type predicts PPM after valve surgery. This score may be useful in the perioperative management of valve surgery patients. PMID- 12628726 TI - Relationship of slow conduction detected by pace-mapping to ventricular tachycardia re-entry circuit sites after infarction. AB - OBJECTIVES: This study sought to characterize the relationship of conduction delays detected by pace-mapping, evident as a stimulus to QRS interval (S-QRS) delay >or=40 ms, to ventricular tachycardia (VT) re-entry circuit isthmuses defined by entrainment and ablation. BACKGROUND: Areas of slow conduction and block in old infarcts cause re-entrant VT. METHODS: In 12 patients with VT after infarction, pace-mapping was performed at 890 sites. Stimulus to QRS intervals were measured and plotted in three-dimensional reconstructions of the left ventricle. Conduction delay was defined as >or=40 ms and marked delay as >80 ms. The locations of conduction delays were compared to the locations of 14 target areas, defined as the region within a radius of 2 cm of a re-entry circuit isthmus. RESULTS: Pacing captured at 829 sites; 465 (56%) had no S-QRS delay, 364 (44%) had a delay >or=40 ms, and 127 (15%) had a delay >80 ms. Sites with delays were clustered in 14 discrete regions, 13 of which overlapped target regions. Only 1 of the 14 target regions was not related to an area of S-QRS delay. Sites with marked delays >80 ms were more often in the target (52%) than sites with delays 40 to 80 ms (29%) (p < 0.0001). CONCLUSIONS: Identification of abnormal conduction during pace-mapping can be used to focus mapping during induced VT to a discrete region of the infarct. Further study is warranted to determine if targeting regions of conduction delay may allow ablation of VT during stable sinus rhythm without mapping during VT. PMID- 12628727 TI - Identification of acutely ischemic myocardium using ultrasonic strain measurements. A clinical study in patients undergoing coronary angioplasty. AB - OBJECTIVES: The goal of this study was to investigate whether the changes in myocardial deformation measured with ultrasonic strain could accurately identify acutely ischemic myocardium during coronary angioplasty. BACKGROUND: Early identification of acute myocardial ischemia has important clinical implications. The accuracy of ultrasonic strain for the detection of acute myocardial ischemia has been validated in animal experiments but has not been investigated in the clinical setting. METHODS: In 73 patients (64 +/- 12 years), either radial or longitudinal strain values were monitored in the "at-risk" segments before, during, and early after right, circumflex, and left anterior descending coronary angioplasty. Based on the visual wall motion assessed before the angioplasty, segments were divided into normokinetic (group I) and hypo/akinetic (group II). Strain data in the "at-risk " segments were compared with values derived from the adjacent nonischemic segments and normal values in 20 controls. RESULTS: Coronary occlusion induced a marked reduction in the systolic strain both in the radial (from 49 +/- 6.9% to 23 +/- 4.6% in group I and from 21.9 +/- 11% to 11.3 +/- 8.4% in group II, p < 0.001) and longitudinal directions. Concomitantly, postsystolic strain increased (from 3.8 +/- 3.1% to 14.6 +/- 9.5% in group I, and from 4.4 +/- 3.7% to 11.3 +/- 7.8% in group II in radial direction, p < 0.001). Upon reperfusion, all deformation parameters returned to near preocclusion values. In comparison with control, baseline, and reperfusion data, the systolic and postsystolic strain parameters measured during total coronary occlusion identified acutely ischemic myocardium with a sensitivity of 86% to 95% and a specificity of 83% to 89%. CONCLUSIONS: In this model of acute ischemia, ultrasonic strain indexes differentiate acutely ischemic segments from both normal and dysfunctional myocardium. This should be a promising new approach to the bedside monitoring of acute ischemic changes in regional myocardial function. PMID- 12628728 TI - Peak early diastolic mitral annulus velocity by tissue Doppler imaging adds independent and incremental prognostic value. AB - OBJECTIVES: The aim of this study was to ascertain if left ventricular mitral annulus velocities measured by tissue Doppler imaging (TDI) are more powerful predictors of outcome compared with clinical data and standard Doppler echocardiographic parameters. BACKGROUND: Tissue Doppler imaging of basal or mitral annulus velocities provides rapid assessment of ventricular long axis function. But it is not known if TDI-derived velocities in systole and diastole add incremental value and are superior to the standard Doppler-echocardiographic measurements as a predictor of outcome. METHODS: The study population consisted of 518 subjects, 353 with cardiac disease and 165 normal subjects who had full Doppler two-dimensional-echocardiographic studies with measurement of mitral inflow velocities in early and late diastole, E-wave deceleration time (DT), peak systolic mitral annular velocity (Sm) early and late diastolic mitral annular velocity (Em and Am) by TDI, early diastolic flow propagation velocity, and standard chamber dimensions. All subjects were followed up for two years. The end point was cardiac death. RESULTS: Tissue Doppler imaging mitral annulus systolic and diastolic velocities were all significantly lower in the non-survivors (all p < 0.05) as was DT (p = 0.024). In the Cox model the best predictors of mortality were Em, Sm, Am, left ventricular ejection fraction, left ventricular mass, and left atrial diameter in systole (LADs). By backward stepwise analysis Em and LADs were the strongest predictors. After forcing the TDI measurements into the covariate model with clinical and mitral DT <0.16 s, Em provided significant incremental value for predicting cardiac mortality (p = 0.004). CONCLUSIONS: Mitral annulus velocity measured by TDI in early diastole gives incremental predictive power for cardiac mortality compared to clinical data and standard echocardiographic measurements. This easily available measurement adds significant value in the clinical management of cardiac patients. PMID- 12628729 TI - Detection of myocardial viability by contrast echocardiography in acute infarction predicts recovery of resting function and contractile reserve. AB - OBJECTIVES: We sought to determine whether myocardial contrast echocardiography (MCE) performed before and early after primary coronary stenting (PCS) in patients with acute myocardial infarction (AMI) could predict recovery of resting left ventricular systolic function and contractile reserve. BACKGROUND: Myocardial contrast echocardiography can be used to assess perfusion within the risk area before PCS and the extent of necrosis soon after PCS. METHODS: In 30 patients with AMI, MCE and two-dimensional echocardiography were performed before PCS and 3 to 5 days and 4 weeks after PCS. Contractile reserve was assessed by dobutamine echocardiography at four weeks in patients with persistent severe wall motion abnormalities. RESULTS: Of segments without perfusion at 3 to 5 days, 95% had severe hypokinesis to akinesis at 4 weeks. Of segments with normal perfusion at 3 to 5 days, 90% had normal wall motion or mild hypokinesis at 4 weeks, whereas those with partial perfusion at 3 to 5 days were evenly divided between normal wall motion, hypokinesis, and akinesis. In segments with persistent severe wall-motion abnormalities at four weeks, contractile reserve was found in >80% of segments with perfusion, compared with only 10% of segments without detectable perfusion (p < 0.01). The presence of myocardial perfusion by MCE before PCS was associated with maintained or improved perfusion at 3 to 5 days and eventual recovery of resting wall motion. CONCLUSIONS: Myocardial contrast echocardiography performed early after PCS provides information on the extent of infarction, and hence the likelihood for recovery of resting systolic function or contractile reserve. The presence of perfusion before PCS, from either collateral or antegrade flow, predicts the maintenance of perfusion and recovery of systolic function. PMID- 12628730 TI - Susceptibility-sensitive magnetic resonance imaging detects human myocardium supplied by a stenotic coronary artery without a contrast agent. AB - OBJECTIVES: Evaluation of the severity of a coronary artery stenosis is of paramount importance for therapy. A relevant stenosis provokes post-stenotic microvascular dilation with capillary recruitment. This autoregulatory response was investigated in the present study by use of susceptibility-sensitive magnetic resonance imaging (MRI) without contrast agents. BACKGROUND: Functional alterations of the microvascular system may be studied noninvasively and without a contrast agent by susceptibility-sensitive MRI, which is based on the paramagnetic property of deoxyhemoglobin. This effect, also referred to as the "blood oxygenation level-dependent (BOLD) effect," is investigated by phase relaxation (T(2)*) measurements. METHODS: In patients (n = 16) with single-vessel coronary artery disease, no history of myocardial infarction, normal left ventricular function at rest, and a positive stress echocardiogram, the susceptibility-sensitive parameter T(2)* was assessed in the myocardium. RESULTS: In regions associated with the stenotic artery, T(2)* was significantly lower than in residual myocardium (p < 0.01). This difference in T(2)* increased after application of the vasodilator dipyridamole (p < 0.001). In patients being re investigated after therapeutic interventions, the microvascular dilation was partly removed. CONCLUSIONS: For the first time, we could show that myocardial BOLD MRI detects post-stenotic capillary recruitment dependent on coronary artery stenosis. PMID- 12628731 TI - Testing coronary flow reserve without a provocative stress. A "BOLD" approach. PMID- 12628733 TI - Hypothermic circulatory arrest for the surgical treatment of complicated adult coarctation of the aorta. AB - OBJECTIVES: This study was designed to evaluate the surgical treatment of recurrent coarctation by a new technique. BACKGROUND: Recurrent coarctation either from aneurysm or recurrent constriction is a difficult problem in the adult because of the possible interruption of important collateral circulation. METHODS: We reviewed four patients who underwent recurrent coarctation surgery with the use of deep hypothermic circulatory arrest (HCA). RESULTS: All four patients survived. Deep HCA facilitated precise surgical resection and there was no postoperative paraplegia, stroke, or myocardial infarction. CONCLUSIONS: Deep HCA and resection and grafting of the coarctation is indicated for complicated adult coarctations, particularly when the collateral circulation is in doubt. PMID- 12628732 TI - Electromechanical mapping versus positron emission tomography and single photon emission computed tomography for the detection of myocardial viability in patients with ischemic cardiomyopathy. AB - OBJECTIVES: We compared catheter-based electromechanical mapping (NOGA system, Biosense-Webster, Haifa, Israel) with positron emission tomography (PET) and single photon emission computed tomography (SPECT) for prediction of reversibly dysfunctional myocardium (RDM) and irreversibly dysfunctional myocardium (IDM) in patients with severe left ventricular dysfunction. Furthermore, we established the optimal discriminatory value of NOGA measurements for distinction between RDM and IDM. BACKGROUND: The NOGA system can detect viable myocardium but has not been used for prediction of post-revascularization contractile function in patients with ischemic cardiomyopathy. METHODS: Twenty patients (19 males, age [mean +/- SD] 60 +/- 16 years, ejection fraction [EF] 29 +/- 6%) underwent viability testing with NOGA and PET or SPECT before revascularization. Left ventricular function was studied at baseline and six months after revascularization. RESULTS: The EF increased to 34 +/- 13% at six months (p < 0.05 vs. baseline). The 58 RDM and 57 IDM regions differed with regard to unipolar voltage amplitude (UVA) (9.2 +/- 3.9 mV vs. 7.6 +/- 4.0 mV, p < 0.05), normalized UVA (106 +/- 54% vs. 75 +/- 39%, p < 0.05), and tracer uptake (76 +/- 17% vs. 60 +/- 20%, p < 0.05). The NOGA local shortening did not distinguish between RDM and IDM (6.4 +/- 5.8% vs. 5.4 +/- 6.6%). By receiver operating characteristic curve analysis, myocardial tracer uptake had better diagnostic performance than UVA (area under curve [AUC] +/- SE: 0.82 +/- 0.04 vs. 0.63 +/- 0.05, p < 0.05) and normalized UVA (AUC +/- SE: 0.70 +/- 0.05, p < 0.05). Optimal threshold was defined as the value yielding sensitivity = specificity for prediction of RDM. Sensitivity and specificity were 59% at a UVA of 8.4 mV, 65% at a normalized UVA of 83%, and 78% at a tracer uptake of 69%. CONCLUSIONS: The NOGA system may discriminate RDM from IDM with optimal discriminatory values for UVA and normalized UVA of 8.4 mV and 83%, respectively. However, the diagnostic performance does not reach the level obtained by PET and SPECT in patients with severe heart failure. PMID- 12628734 TI - Echocardiographic predictors of aortopulmonary collaterals in infants with tetralogy of fallot and pulmonary atresia. AB - OBJECTIVES: This study was designed to identify echocardiographic predictors of aortopulmonary collaterals (APCs) in infants with tetralogy of Fallot and pulmonary atresia (TOF/PA) and to prospectively validate these predictors. BACKGROUND: In patients with TOF/PA, routine preoperative diagnostic cardiac catheterization is usually performed to determine the presence and distribution of APCs and the anatomy of the branch pulmonary arteries. METHODS: The clinical and diagnostic imaging data of infants with TOF/PA treated between 1990 and 2000 were reviewed. Patients were classified into two groups based on cineangiographic findings: 1) no APCs (n = 34), and 2) >or=1 APCs (n = 59). Echocardiographic variables were examined for their ability to identify patients having >or=1 APCs. RESULTS: Median branch pulmonary artery diameter Z scores were significantly larger in patients without APCs compared with those having >or=1 APCs: -0.56 versus -3.24 for the left pulmonary artery and -0.76 versus -3.46 for the right pulmonary artery (p < 0.001). The presence of a branch pulmonary artery diameter Z score or=1 APCs. Detection of APCs by color Doppler was 93% sensitive and 91% specific. A combination of branch pulmonary artery diameter Z score or=1 APCs. The diagnostic accuracy of these echocardiographic variables was subsequently validated in a prospective study of 11 infants. CONCLUSIONS: Echocardiography is a sensitive and specific test for the detection of >or=1 APCs in infants with TOF/PA. These data can be used to select patients who can undergo complete repair of TOF/PA without further preoperative diagnostic imaging. PMID- 12628735 TI - Molecular mechanisms of early electrical remodeling: transcriptional downregulation of ion channel subunits reduces I(Ca,L) and I(to) in rapid atrial pacing in rabbits. AB - OBJECTIVES: The purpose of the study was to characterize the ionic and molecular mechanisms in the very early phases of electrical remodeling in a rabbit model of rapid atrial pacing (RAP). BACKGROUND: Long-term atrial fibrillation reduces L type Ca(2+) (I(Ca,L)) and transient outward K(+) (I(to)) currents by transcriptional downregulation of the underlying ionic channels. However, electrical remodeling starts early after the onset of rapid atrial rates. The time course of ion current and channel modulation in these early phases of remodeling is currently unknown. METHODS: Rapid (600 beats/min) right atrial pacing was performed in rabbits. Animals were divided into five groups with pacing durations between 0 and 96 h. Ionic currents were measured by patch clamp techniques; messenger ribonucleic acid (mRNA) and protein expression were measured by reverse transcription-polymerase chain reaction and Western blot, respectively. RESULTS: L-type calcium current started to be reduced (by 47%) after 12 h of RAP and continued to decline as pacing continued. Current changes were preceded or paralleled by decreased mRNA expression of the Ca(2+) channel beta subunits CaB2a, CaB2b, and CaB3, whereas significant reductions in the alpha(1) subunit mRNA and protein expression began 24 h after pacing onset. Transient outward potassium current densities were not altered within the first 12 h, but after 24 h, currents were reduced by 48%. Longer pacing periods did not further decrease I(to). Current changes were paralleled by reduced Kv4.3 mRNA expression. Kv4.2, Kv1.4, and the auxiliary subunit KChIP2 were not affected. CONCLUSIONS: L-type calcium current and I(to) are reduced in early phases of electrical remodeling. A major mechanism appears to be transcriptional downregulation of underlying ion channels, which partially preceded ion current changes. PMID- 12628736 TI - Amlodipine inhibits doxorubicin-induced apoptosis in neonatal rat cardiac myocytes. AB - OBJECTIVES: We examined whether amlodipine, a calcium channel antagonist with potent antioxidant activity, inhibits doxorubicin-induced apoptosis in cultured neonatal rat cardiac myocytes. BACKGROUND: Recent studies have shown that doxorubicin induces apoptosis as well as necrosis in myocytes through generation of reactive oxygen species. METHODS: The effects of amlodipine and several other antioxidants on doxorubicin-induced oxidative stress and mitochondria-mediated apoptosis were examined. RESULTS: Treatment of myocytes with doxorubicin (10(-6) mol/l) for 14 h increased the number of cells with elevated peroxides, as histochemically estimated by 2',7'-dichlorofluorescin (DCF) diacetate, and the percentage of apoptotic myocytes, as estimated by Hoechst 33258 nuclear staining, compared with control myocytes (25.0 +/- 1.6% vs. 5.2 +/- 1.2%). Moreover, doxorubicin-induced myocyte apoptosis was also confirmed by annexin V-fluorescein isothiocyanate binding assay. Doxorubicin induced a reduction in myocyte adenosine 5'-triphosphate content, a loss of mitochondrial membrane potential, cytochrome c release from the mitochondria into the cytosol, and caspase-3 activation to 1.9-fold of control. Amlodipine significantly attenuated increased DCF fluorescence, inhibited the mitochondria-mediated apoptotic responses described earlier, and decreased apoptosis in the doxorubicin-treated myocytes in a dose-dependent fashion. Amlodipine at 10(-6) mol/l significantly decreased apoptosis to 15.4 +/- 0.7%, and this antiapoptotic action was more effective than that seen with other antioxidants, including probucol, ascorbic acid, and alpha tocopherol. In contrast, the calcium channel antagonist nifedipine (10(-6) mol/l) did not inhibit apoptosis. Catalase, glutathione, and N-acetylcysteine, but not mannitol or superoxide dismutase, significantly decreased DCF fluorescence and attenuated myocyte apoptosis induced by doxorubicin to 18.7 +/- 1.2%, 19.1 +/- 1.7%, and 18.7 +/- 0.6%, respectively. CONCLUSIONS: Amlodipine significantly inhibits doxorubicin-induced myocyte apoptosis by suppressing the mitochondrial apoptotic pathway. This effect is attributed to the antioxidant properties of amlodipine, affecting mainly hydrogen peroxide. PMID- 12628738 TI - The physician and war. PMID- 12628737 TI - Autologous skeletal myoblasts transplanted to ischemia-damaged myocardium in humans. Histological analysis of cell survival and differentiation. AB - OBJECTIVES: We report histological analysis of hearts from patients with end stage heart disease who were transplanted with autologous skeletal myoblasts concurrent with left ventricular assist device (LVAD) implantation. BACKGROUND: Autologous skeletal myoblast transplantation is under investigation as a means to repair infarcted myocardium. To date, there is only indirect evidence to suggest survival of skeletal muscle in humans. METHODS: Five patients (all male; median age 60 years) with ischemic cardiomyopathy, refractory heart failure, and listed for heart transplantation underwent muscle biopsy from the quadriceps muscle. The muscle specimen was shipped to a cell isolation facility where myoblasts were isolated and grown. Patients received a transplant of 300 million cells concomitant with LVAD implantation. Four patients underwent LVAD explant after 68, 91, 141, and 191 days of LVAD support (three transplant, one LVAD death), respectively. One patient remains alive on LVAD support awaiting heart transplantation. RESULTS: Skeletal muscle cell survival and differentiation into mature myofibers were directly demonstrated in scarred myocardium from three of the four explanted hearts using an antibody against skeletal muscle-specific myosin heavy chain. An increase in small vessel formation was observed in one of three patients at the site of surviving myotubes, but not in adjacent tissue devoid of engrafted cells. CONCLUSIONS: These findings represent demonstration of autologous myoblast cell survival in human heart. The implanted skeletal myoblasts formed viable grafts in heavily scarred human myocardial tissue. These results establish the feasibility of myoblast transplants for myocardial repair in humans. PMID- 12628739 TI - Prehospital thrombolytic therapy for ST-segment elevation myocardial infarction. PMID- 12628741 TI - Proceedings of the VI Workshop on Apoptosis in Biology and Medicine. 25-29 May 2002, Calabria, Italy. PMID- 12628742 TI - The ASPP family: deciding between life and death after DNA damage. AB - It is well established that p53 is a primary target for mutation in human cancer. p53 carries out the important task of ensuring that damaged DNA is not passed on during cell division, a duty that it performs by either inhibiting the cell cycle or inducing apoptosis. However, it is unclear how this decision is made. The recent identification of the ASPP family of proteins, which act to direct the cell away from cell cycle arrest and towards death following p53 upregulation, may explain how this dilemma is resolved. Furthermore, the observation that ASPP2 is in fact the full length form of the previously identified 53BP2/Bbp protein has clarified the ambiguous data that has been generated in relation to this molecule. The further characterisation of these proteins will enable us to gain further insights into the response of the cell to DNA damage and the progression of the cell towards malignancy. PMID- 12628743 TI - TRAIL-induced signalling and apoptosis. AB - Prominent among cell surface molecules capable of initiating apoptosis are members of the tumour necrosis factor (TNF) family of ligands and receptors. Death-inducing members of the TNF receptor family each contain a cytoplasmic 'death domain' (DD): a protein-protein interaction motif critical for engaging downstream components of the signal transduction machinery. Following activation of these receptors by their cognate ligands, a death-inducing signalling complex (DISC) is formed by recruitment of cytoplasmic DD-containing proteins to the receptor DD. The best-studied death-inducing ligand-receptor pairs are TNF/TNF receptor-1 (TNFR1) and CD95L/CD95 (Fas, Apo-1). A more recently identified member of the TNF ligand family is TNF-related apoptosis-inducing ligand (TRAIL), Apo 2L, which induces apoptosis in a number of tumour cell lines, yet is relatively non-toxic to normal cells. Consistent with its lack of toxicity, TRAIL is constitutively expressed in many human tissues. This raises the conundrum of why tumour cells are sensitive to TRAIL-mediated apoptosis and normal tissues resistant. One possibility lies in the existence of a family of four membrane bound TRAIL receptors (TRAIL-R1-R4), which although able to bind TRAIL, differ in their ability to transduce the death signal. Recently, this hypothesis has been challenged based on poor correlations between TRAIL receptor expression and TRAIL sensitivity. This suggests that other factors, such as TRAIL-induced NF-kappaB activation or death inhibitors including c-FLIP, are involved in determining differential sensitivity to TRAIL. We have investigated TRAIL sensitivity in primary tumour cells together with TRAIL-induced signalling pathways in both sensitive and resistant cell lines. Our results and their implications for the potential use of TRAIL in cancer therapy are discussed. PMID- 12628745 TI - Superoxide as inhibitor of calcineurin and mediator of redox regulation. AB - The concept of NO as a redoxactive messenger has to be broadened by including superoxide as an antagonistic messenger. Superoxide alone was found to inhibit calcineurin by interacting with the FeII-ZnII binuclear site. This links oxidative stress conditions with a Ca-dependent phosphorylation/dephosphorylation cascade. When NO and superoxide are generated at equal fluxes the resulting peroxynitrite can cause tyrosine nitrations (e.g. prostacyclin synthase inhibition) or oxidations of zinc-fingers in proteins, indicating a new messenger function. Finally, if generated in excess, NO can convert peroxynitrite to N(2)O(3) as a nitrosating agent. Thus, the NO/superoxide system provides four different messengers affecting important regulatory pathways. PMID- 12628744 TI - Involvement of DNA damage and repair systems in neurodegenerative process. AB - The study summarizes some recent data from our and other groups underlining the contribution to neurodegeneration of two transcription factors known to be involved in DNA damage sensing and repairing: the tumour suppressor gene p53 and the component of the DNA repair system MSH2. Both proteins participate in the cancer prevention machinery for the body as well as in the neurodegenerative process, suggesting that cancer and neurodegenerative disease may share common genetic risk factors for the development and progression of the disease. Here we show that, in neuronal cells, divergent cellular insults, i.e. the exposure to glutamate, beta-amyloid (Abeta) or H(2)O(2), may converge to a common pathway that initiate with elevation of p53 protein levels. We also found that in SH-SY5Y neuronal cells H(2)O(2) induced the activation of DNA repair system with the nuclear translocation of MSH2, and PCNA. Differently no changes in MSH2 and PCNA cellular distribution were found in undifferentiating SH-SY5Y cells exposed to H(2)O(2). This argues that defects in the repair of, or response to, DNA damage impact significantly on brain function. PMID- 12628746 TI - Redox control of hepatic cell death. AB - In necrotic liver failure like upon acetaminophen overdose, loss of the major intracellular thiol antioxidant glutathione was shown to be causal for hepatic dysfunction. In sharp contrast, fulminant apoptotic liver destruction upon overstimulation of the death receptors TNFR1 and CD95 was not associated with reduced hepatic glutathione levels. In view of the importance of the role of reactive oxygen intermediates versus antioxidants for apoptosis, we investigated the effect of phorone-induced enzymatic GSH depletion on the sensitivity of the liver towards CD95- or TNFR1-mediated hepatotoxicity. Our findings demonstrate in vivo that receptor-mediated hepatic apoptosis is disabled when glutathione is depleted, i.e. that an intact glutathione status is a critical determinant for the execution of apoptosis. In vitro, we did mechanistic studies in lymphoid cell lines and found that pro-caspase-8 at the CD95 death receptor and the mitochondrial activation of pro-caspase-9 are the enzyme targets that require sufficient intracellular reduced glutathione for their activation. PMID- 12628747 TI - Nitric oxide evoked p53-accumulation and apoptosis. AB - The tumor suppressor p53 accumulates under conditions of cellular stress and affects cell cycle progression and/or apoptosis. This has been exemplified for endogenously produced or exogenously supplied nitric oxide (NO) and thus accounts at least in part for cell destructive signaling qualities of this bioactive molecule and/or derived reactive nitrogen species. However, detailed mechanisms of toxicity and pathways of cell demise remain to be elucidated. Establishing that NO-treatment left the ubiquitination and the p53-Mdm2 interaction intact may point to an impaired nuclear-cytoplasmic shuttling to account for p53 stabilization. This was verified by heterokaryon analysis. We conclude that attenuated nuclear export contributes to stabilization and activation of p53 under the influence of NO. PMID- 12628748 TI - Apoptosis induced in neuronal cells by oxidative stress: role played by caspases and intracellular calcium ions. AB - Reactive oxygen species (ROS) have been implicated in the pathophysiology of many neurologic disorders and brain dysfunction. In the same pathological settings evidence has been provided in favour of a participation of intracellular Ca(2+) concentration altered homeostasis in the chain of events leading to neuronal apoptosis. In the present review literature reports and experimental data on the relationship between caspase activation and alteration of intracellular calcium concentrations in the mechanisms triggering neuronal apoptosis are discussed. The data gathered support the conclusion that during oxidative stress in neuronal cells the production of ROS triggers a mechanism that, through the release of cytochrome c from mitochondria and caspase-3 activation, leads to apoptosis; the concomitant ROS-mediated elevation of intracellular Ca(2+) concentration triggers caspase-2 activation but both events do not seem to be involved in cell death. PMID- 12628749 TI - Apoptotic mechanisms and antiapoptotic therapy in the MPTP model of Parkinson's disease. AB - The 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) model constitutes the best-characterized toxin paradigm for Parkinson's disease, faithfully replicating most of its clinical and pathological hallmarks. Many lines of evidence point to a significant contribution of apoptosis to cell death after application of 1 methyl-4-phenylpyridinium (MPP(+)) in cell culture or MPTP in vivo. This holds true for apoptotic DNA strand breaks, activation of the JNK pathway and caspases, induction of Par-4 protein and the protection conferred by interference with p53, Apaf-1 or Bax signalling. In MPTP models, intervention in upstream events of apoptosis, e.g. by inhibition of the JNK pathway, provides morphological and functional rescue. In contrast, inhibition of the propagation and execution phase of apoptosis, e.g. by inhibition of caspases, blocks or delays cell death but may not recover neuronal function. At this stage, the combination of an anti apoptotic together with a neurorestorative therapy may be promising. PMID- 12628750 TI - Poly(ADP-ribose) polymerase as a key player in excitotoxicity and post-ischemic brain damage. AB - Poly(ADP-ribose) polymerases (PARPs) are a group of protein-modifying and nucleotide-polymerizing enzymes able to catalyze the transfer of multiple ADP ribose units from NAD to substrate proteins. In the human genome, 16 different genes encoding for members of this emerging family of enzymes have been identified. Known family members are PARP-1, PARP-2, PARP-3, vPARP, tankyrase 1 and tankyrase 2, each of them with a possible specific role in cell biology. The most studied member of the family is PARP-1, which is abundantly present in the nucleus and is involved in the maintenance of genomic stability. In pathological conditions, highly reactive radical species may cause DNA damage and PARP-1 hyperactivation. This may lead to necrotic cell death through massive NAD consumption. We show that following middle cerebral artery occlusion, rats treated with PARP inhibitors displayed reduced brain infarct volumes. Similarly, PARP inhibitors reduced neuronal death induced by oxygen-glucose deprivation (OGD) or excitotoxins in primary cultures of murine cortical cells. On the contrary, PARP inhibitors did not attenuate the OGD-induced selective loss of CA1 pyramidal cells in rat organotypic hippocampal slices. In addition, they were not neuroprotective against transient bilateral carotid occlusion in gerbils. We observed that post-ischemic brain damage was predominally necrotic in cultured cortical cells, whereas a caspase-dependent apoptotic process was responsible for the CA1 pyramidal cell loss in hippocampal slices. Hence, it appears reasonable to propose PARP inhibitors as useful therapeutic agents in pathological brain conditions were necrosis predominates. PMID- 12628751 TI - Multifaceted roles of nitric oxide in the lateral geniculate nucleus: from visual signal transduction to neuronal apoptosis. AB - The lateral geniculate nucleus (LGN) is the thalamic relay of retinal inputs to the visual cortex. It contains a rich array of brain terminals, which modulate the visual signals to the cortex. Several data have documented that beside cholinergic, GABA-nergic, istaminergic, serotoninergic, and glutamatergic signals, the LGN contains also fibers and interneurons expressing the enzyme that produces nitric oxide (NO). Here, we review the documented physiological roles of NO in the transmission of visual inputs to the cortex and in the processes of activity-dependent refinement of LGN connections. Moreover we focus on the recently suggested role of NO in processes of neurotoxicity in the LGN. Particular relevance is given to studies documenting that, through an excitotoxic cascade, NO triggers apoptosis in the LGN of new-born rats deprived of vision in one eye. Data are also discussed on a possible role of NO in the mechanisms of LGN neuronal loss induced by glaucoma. We believe that a better understanding of the role of NO in the LGN may contribute to discover new experimental strategies for the treatment of degenerative ophthalmic diseases. PMID- 12628752 TI - Dexamethasone-induced thymocytes apoptosis requires glucocorticoid receptor nuclear translocation but not mitochondrial membrane potential transition. AB - Dexamethasone (DEX), a glucocorticoid hormone (GCH) with specificity for the glucocorticoid receptor (GR) induces T lymphocyte and thymocyte apoptosis. DEX activated thymocyte apoptosis requires a sequence of biochemical events including mRNA and protein synthesis. In particular, GCH treatment induces non-genomic mechanisms, such as for example Ca(2+) mobilization and PI-PLC activation, and genomic mechanisms. Most of these events, including protein synthesis, are required and precede caspase activation. As protein synthesis is required for caspases and apoptosis activation, DEX-induced GR nuclear translocation is necessary for apoptosis. Cell treatment with geldanamycin (GA) inhibits the GR nuclear translocation and consequently, caspases activation and apoptosis. Although DEX treatment induces loss of mitochondrial membrane potential (deltapsim) and cytochrome c release, deltapsim induction does not correlate with thymocyte apoptosis. In fact, while Cyclosporin-A and the caspase-9 inhibitor, Z LEHD-FMK, inhibit DEX-induced deltapsim, do not influence apoptosis. These data indicate many biochemical events and are activated by DEX treatment of thymocytes and some, but not all, are required for apoptosis. PMID- 12628753 TI - Chemokines and their receptors in the CNS: expression of CXCL12/SDF-1 and CXCR4 and their role in astrocyte proliferation. AB - The study of chemokine role in the CNS indubitably represents an important step to understanding many aspects of brain pathology, physiology and development. Here we discuss our recent research on the expression of chemokines and chemokine receptors in brain tissues and in cultured CNS cells, with particular regard to the CXCL12/SDF-1-CXCR4 system. We showed their expression in both glial and neuronal cells in basal conditions and their modulation upon stimulation. We demonstrated that CXCL12/SDF-1 in vitro act as a growth factor for astrocytes by stimulating their proliferation, a phenomenon that could represent the basis of pathological conditions such as gliosis and malignant transformation. We investigated the signal transduction pathways, identifying in the sequential activation of G-protein-PI-3Kinase-ERK1/2 the main signaling cascade linked to the CXCL12/SDF-1-induced proliferation in astrocytes. PMID- 12628754 TI - Regulation of NO-dependent cyclic GMP formation by inflammatory agents in neural cells. AB - In the CNS, NO is an important physiological messenger involved in the modulation of brain development, synaptic plasticity, neuroendocrine secretion, sensory processing, and cerebral blood flow [Annu. Rev. Physiol. 57 (1995) 683]. These NO actions are largely mediated by cyclic GMP (cGMP) formed by stimulation of soluble guanylyl cyclase (sGC). NO has also been recognized as a neuropathological agent in conditions such as epilepsy, stroke and neurodegenerative disorders. In these conditions, NO may contribute to excitotoxic cell death and neuroinflammatory cell damage [Brain Res. Bull. 41 (1996) 131; Glia 29 (2000) 1]. NO can be formed in every type of CNS parenchymal cell, however, cGMP appears to be formed mainly in neurons and astroglia [Annu. Rev. Physiol. 57 (1995) 683]. There is a large body of information about the regulation of NO formation in brain cells under both normal and pathological conditions but much less is known about the control of cGMP generation, in particular during neuroinflammation when there is a high NO output. Here we briefly review our present knowledge on the regulation of NO-dependent cGMP formation in brain cells under inflammatory conditions. PMID- 12628755 TI - Neurobiological mediators of neuronal apoptosis in experimental neuroAIDS. AB - Neuronal loss has often been described at post-mortem in the brain neocortex of patients suffering from AIDS. Neuroinvasive strains of HIV infect macrophages, microglial cells and multinucleated giant cells but not neurones. Processing of the virus by cells of the myelomonocytic lineage yields viral products that, in conjunction with potentially neurotoxic molecules generated by the host, might initiate a complex network of events which leads neurones to death. In particular, the HIV-1 coat glycoprotein gp120 has been proposed as a likely aetiologic agent of the described neuronal loss because it causes death of neurones in culture. More recently, it has been shown that brain neocortical cell death is caused in rat by intracerebroventricular injection of a recombinant gp120 coat protein and this occurs via apoptosis. The latter observation broadens our knowledge in the pathophysiology of the reported neuronal cell loss and opens a new lane of experimental research for the development of novel therapeutic strategies to limit damage to the brain of patients suffering from HIV associated dementia. PMID- 12628756 TI - Evidence for a role of protein tyrosine kinases in cell death induced by gp120 in CHP100 neuroblastoma cells. AB - HIV-1 coat protein gp120 is able to kill neuronal cells in culture. Here we address the possible role of protein tyrosine kinases (PTKs) in gp120-induced neurotoxicity using the CHP100 human neuroblastoma cell line as experimental model. For this purpose, the effect of specific PTK inhibitors like genistein, herbimycin A and lavendustin A was evaluated on CHP100 cell death elicited by the viral protein. Here we report that genistein (1-10 microM) significantly reduced the cytotoxic effects induced by gp120 (10 pM). The same protective action was offered by a pre-treatment with herbimycin A (0.1-1 microM) or lavendustin A (1 10 microM). Conversely, daidzein (1-100 microM), a genistein analogue devoid of PTK inhibitory properties, failed to reduce CHP100 cell death caused by gp120. These findings suggest that PTKs can be involved in the signal transduction cascade by which the glycoprotein induces neurotoxicity. PMID- 12628758 TI - To die or to sleep, perhaps to dream. AB - Establishing social contacts is the raison d'etre of neurons throughout their entire life span. To form and retain functional connections, neuronal differentiation and death are ruthlessly regulated in development and kept strictly under control in post-mitotic systems. Derangements in neural networks affect neuronal populations at large. Therefore, failure to retain synaptic connectivity is linked to dysfunction and often followed by neuronal death. Loss of neurons is a predominant feature of neurodegenerative disease. Nevertheless, neuronal cell death is not an obligate requirement for neural dysfunction at the level of distributed circuits or local circuits. Although more or less wide spread neuronal loss can occur after acute insults such as brain ischemia or invasion of the brain by pathogens, neuronal death is a hallmark of end-stage neurodegenerative and psychiatric disease. The relative contributions made by loss of synaptic connectivity versus cell death for these diseases are still debated. Here these processes are discussed in relation to acute and chronic CNS disorders. PMID- 12628757 TI - Caspase-1 inhibitors abolish deleterious enhancement of COX-2 expression induced by HIV-1 gp120 in human neuroblastoma cells. AB - The human CHP100 neuroblastoma cell line has been shown to provide an useful in vitro model to elucidate the mechanisms underlying HIV-1 gp120 neurotoxicity. Here we report western blotting evidence demonstrating that exposure to a cytotoxic concentration of the viral coat protein up-regulates expression of the inducible isoform of cyclooxygenase (COX-2) in neuroblastoma cells and this seems to be due to the previously observed increase in secreted IL-1beta. In fact, here we show that acetyl-Tyr-Val-Ala-Asp-chloromethylketone (Ac-YVAD-CMK) and t butoxycarbonyl-L-aspartic acid benzyl ester-chloromethylketone (Boc-Asp-(OBzl) CMK), two inhibitors of Interleukin-1 Converting Enzyme (ICE; also referred to as caspase-1), abolish COX-2 expression enhanced by gp120 and consequent cell death. In addition, NS-398, a selective inhibitor of COX-2 activity, affords neuroprotection strengthening the role of COX-2 in the mechanisms of death. In conclusion, the present data support the notion that IL-1beta is the signal through which gp120 elevates COX-2 expression and the latter is strongly implicated in the mechanisms underlying cytotoxicity. PMID- 12628759 TI - A Leishmania infantum multi-component antigenic protein mixed with live BCG confers protection to dogs experimentally infected with L. infantum. AB - The capacity of a quimeric protein, formed by the genetic fusion of five antigenic determinants from four Leishmania proteins, formulated with BCG, to protect dogs against Leishmania infantum infection is described. The data showed that after i.v. administration of 500,000 parasites of the L. infantum M/CAN/ES/96/BCN150 strain, zymodeme MON-1, the animals became infected as suggested by the humoral response against the parasite antigens. All control unvaccinated dogs had parasites in the lymph nodes at day 150 post-infection. One of these unvaccinated infected dog was parasite negative at day 634 behaving, thus, as resistant. In contrast, only 50% of the immunized dogs had parasites in the lymph nodes at day 150 post-infection. Four of these dogs became parasite negative by day 634 post-infection. The control animals developed at various times during the follow-up period clinical symptoms associated with Leishmaniasis. The control diseased dogs developed also in the liver and spleen some of the abnormal histological features associated with natural visceral Leishmaniasis. The immunized dogs, however, were not only normal at the clinical but also at the anatomo-pathological level. A positive delayed type hypersensitivity (DTH) response was observed in nine of the immunized protected dogs. The data indicated that Q+BCG confers 90% protection against infection and at least 90% protection at the clinical level. PMID- 12628761 TI - Effect of maternally derived antibodies on the clinical signs and immune response in pigs after primary and secondary infection with an influenza H1N1 virus. AB - The aim of this study was to determine the role of maternally derived antibodies (MDA) against an influenza H1N1 virus in the clinical protection of piglets and especially their effect on the development of the active immunity after an infection with a homologous influenza H1N1 virus. Twenty piglets with MDA and 10 piglets without MDA were housed together and inoculated twice with influenza H1N1 virus, at 7 and 15 weeks of age. Nine piglets without MDA were added to these groups at 12 weeks of age to be inoculated at 15 weeks of age only. Clinical signs, body temperature, growth performance, virus excretion, antibody responses, and influenza-specific T-cell response were monitored. It was shown that MDA protect piglets against the clinical consequences of a primary influenza infection, but that this protection is not complete. A short but significant rise in body temperature was observed and growth seemed to be inhibited due to the infection. Piglets with MDA shed virus for a longer period after an infection than piglets without MDA. Piglets with and without MDA were protected against the clinical consequences of a secondary infection. However, both after primary and secondary infection significant differences in immune responses were observed that indicated that pigs with MDA developed a weaker immunity than pigs without MDA. Furthermore, overall growth performances from weaning to slaughter show a trend in favour of pigs without maternal antibodies, compared to pigs with maternal antibodies, mainly caused by a significant better performance in the second half of the finishing period. The results of this study provide us insight in the role of MDA in clinical protection and their influence on active immunity after an influenza virus infection of pigs. Furthermore, it leads us to the discussion about the profitability of massive sow herd vaccinations in an attempt to increase MDA levels in piglets, taking into account the overall performance of these piglets and the possible effects on antigenic drift. PMID- 12628760 TI - Lack of TNF alpha supports persistence of a plasmid encoding the bovine leukaemia virus in TNF(-/-) mice. AB - Tumour necrosis factor (TNF) is well recognised for its role in mediating innate immune responses. However, the mechanisms of TNF that influence the adaptive immune response to viral infections are poorly understood. Over recent years, there has been evidence to suggest a role for TNF in the early phase of infection of ruminants with bovine leukaemia virus (BLV). In this study, we infected TNF(-/ ) mice with a plasmid encoding infectious BLV to further elucidate the role of TNF in BLV infection. TaqMan quantitative PCR showed that proviral DNA was present in genomic DNA isolated from spleen cells of TNF(-/-) mice 4 weeks post infection, whereas it was not detected in wild-type mice. We were not able to detect differences in serum IgM or IgG levels between the TNF(-/-) and wild-type mice, or antibodies to BLV after this short period. In showing that the lack of TNF enables the plasmid encoded BLV to persist longer, and therefore rendering the mice more susceptible to an infection with BLV, the data suggest an important defence function of TNF in the early phase of BLV infection. PMID- 12628762 TI - Sequencing and analysis of genomic DNA and cDNA encoding TNF-alpha in the squirrel monkey (Saimiri sciureus). AB - If a number of cytokines and growth factors that have been characterized from human cells were investigated in non-human primates, results from such approaches would allow the development of assays to detect and quantitate cytokines in experimental models. Tumor necrosis factor-alpha (TNF-alpha) is an important pluripotent cytokine which plays a crucial role in host defense. As yet, no complete molecular data have been reported for the squirrel monkey TNF-alpha. Polymerase chain reaction (PCR) primers were used to trace introns, by comparing product sizes obtained using cDNA and genomic DNA as templates. The genomic DNA is composed of four exons and three introns with 1793 nucleotides. The corresponding cDNA is 702 nucleotides and phylogenetic analysis showed that the Saimiri sciureus was most closely related to that of the genus Aotus, a new-world primate, compared to old-world primates (genus Macaca and Papio). The deduced TNF alpha protein consists of 233 amino acids with 82% identity to human, 95% to new world monkeys and 79% to old-world monkeys. The cloned TNF-alpha cDNA will be useful to quantitate TNF-alpha at the mRNA level. PMID- 12628763 TI - Monoclonal anti-equine IgE antibodies with specificity for different epitopes on the immunoglobulin heavy chain of native IgE. AB - In this study we describe the generation of monoclonal antibodies (mAbs), which recognize different epitopes of the equine IgE constant heavy chain. Equi-murine recombinant IgE (rIgE), composed of the murine V(H)186.2 heavy chain variable region, linked to the equine IgE constant heavy chain and expressed together with the murine lambda(1) chain in J558L cells was used to immunize BALB/C mice. A total of 17 different mAbs were obtained, which recognized the rIgE heavy chain constant region. None of the mAbs reacted with monoclonal equine isotypes IgM, IgG1 (IgGa), IgG3 (IgG(T)), IgG4 (IgGb) or isolated equine light chains, IgGc and IgA from horse serum, or the native mAb B1-8delta, expressing the same heavy chain variable regions and light chains. One of the mAbs (alphaIgE-132) recognized the recombinant equine IgE, but did not recognize any protein in equine serum, i.e. native IgE. A total of 16 mAbs detected a serum protein of approximately 210,000Da on Western blots, corresponding to the expected MW of native IgE. In addition, one of the mAbs (alphaIgE-176) detected a protein of 76,000Da under reducing conditions, most likely the equine IgE heavy chain. According to binding inhibition studies, the equine IgE specific mAbs recognize at least two different epitopes of the equine IgE. In an ELISA using two anti-IgE mAbs which recognized different epitopes, no significant differences in the concentration of total serum IgE could be detected between adult Icelandic horses with IgE-mediated type I allergy (summer eczema) and healthy control animals. In Icelandic horse foals, no serum IgE could be measured 6 months post partum. All anti-IgE mAbs recognized a small population (1.3+/-0.5%) of leukocytes from adult Icelandic horses by surface immunofluorescence, but no cells could be detected in foal blood. The stained leukocytes from adult horses could be enriched by magnetic cell sorting and contained 32% basophils, 53% monocytes and/or large lymphocytes, 13% small lymphocytes and 2% eosinophils. PMID- 12628764 TI - Induction of lymphopenia and inhibition of T cell function during acute infection of swine with foot and mouth disease virus (FMDV). AB - Foot and mouth disease virus (FMDV) is a picornavirus that causes an acute vesicular disease of cloven-hoofed animals. This virus continues to be a threat to livestock worldwide with outbreaks causing severe economic losses. The present study shows an analysis of immune system phenotype and function during the acute phase of FMDV infection in swine. In the first days of infection, a significant lymphopenia is observed that involves all T cell subsets, CD4(+), CD8(+), and CD4(+)/CD8(+). This marked lymphopenia is not a result of active infection of PBMC with the virus. Further, the response of residual peripheral blood T cells to the mitogen, Concanavalin A (ConA) is significantly reduced and occasionally eliminated. Animals usually resolve clinical signs of disease and develop antigen specific T cell responses to the virus and recover ConA reactivity. These characteristics of acute phase infection likely play an important role in viral pathogenesis, propagation and shedding of viral particles and may be targeted as a way of improving vaccine formulations. PMID- 12628766 TI - Characterisation of humoral immune responses in dogs vaccinated with irradiated Ancylostoma caninum. AB - Infection with Ancylostoma caninum, an intestinal hookworm of dogs, can cause debilitating and potentially life-threatening disease. In the current study, protective immunity to hookworm infection was induced in dogs following vaccination with irradiation-attenuated third-stage larvae (L3) with significant reductions in both worm (P<0.03) and faecal egg counts (P<0.0004) following a challenge infection. Vaccination with irradiated L3 and challenge with infective L3 stimulated a dominant antibody response to antigens of less than 20 kDa in an excretory/secretory extract from adult parasites. Immunoscreening of an adult A. caninum cDNA library with antisera from the vaccine trial identified a number of clones. The three clones with the strongest immunoreactivity proved to be identical and encoded a peptide with similarity to the N-terminal domain of the tissue matrix metalloproteinase inhibitor (TIMP)-2 mammalian tissue metalloproteinase inhibitor family. PMID- 12628767 TI - A review of M. bovis BCG protection against TB in cattle and other animals species. AB - Bovine tuberculosis (TB) causes severe economic losses in livestock due to low production, animal deaths and condemnation of carcasses. It is also an important constraint in international trade of animals and animal products. A scientific committee in Great Britain in 1997 concluded that the development of a cattle vaccine would be the best option for long-term control of TB. However, vaccination of cattle currently is not accepted because the vaccine interferes with the skin reaction to the tuberculin test in the field. Efficacy of M. bovis BCG in protecting bovine and other animal species against tuberculous infection has received much study. Vaccination of cattle prevents the spread of the disease in populations by reducing the number and size of the lesions, and the load of bacteria (rather than by preventing infection). We review the literature about the efficacy of BCG in protecting cattle and other animal species against infection with field strains of M. bovis and discusses its potential use in programs of TB control in high-prevalence populations. PMID- 12628765 TI - Temporal changes in cytokine expression of foals during the first month of life. AB - Foals are uniquely susceptible to a wide variety of opportunistic infections normally associated with immunodeficiencies. Little is understood about the immune system of foals during the neonatal period. An apparent age-related susceptibility predisposes neonatal foals to infectious diseases and hinders therapeutic and preventative interventions for these diseases. Cytokine expression is correlated with the type of immune response as well as the severity of a disease. In this study, we measured foal peripheral blood mononuclear cell (PBMC)-specific mRNA cytokine expression from 72 foals from three different farms during the first 4 weeks of life. Interleukin-1alpha (IL-1alpha), IL-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p35, IL-12p40, interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), and transforming growth factor-beta1 (TGF beta1) were cloned and transcribed in vitro to generate antisense probes for ribonuclease protection assays. Using linear mixed-effect models, we determined that IFN-gamma, TGF-beta1, and IL-1alpha increased significantly (P<0.05) with age. PMID- 12628768 TI - Risk factors associated with BMSCC greater than 200 000 cells/ml in dairy herds in southern Chile. AB - A cross-sectional survey of 523 dairy farms in the south of Chile was carried out to quantify risk factors associated with bulk-milk somatic-cell count (BMSCC) >200 x 10(3)cells/ml. Questionnaires followed by one reminder were sent to 3710 dairy farms via the 11 milk-processing plants that they supplied in October 1998. The response proportion was 14.1%. The median BMSCC was 289 x 10(3) cells/ml (range: 74 x 10(3) to 1800 x 10(3)cells/ml). The median herd size was 70 cows (range: 7-616); herd size was not associated with BMSCC. The annual milk yield of 33.2% of the herds was <4000 l and 53.4% had an annual milk yield of 4 x 10(3) to 6 x 10(3) l. Clinical-mastitis records were kept by 55.3% of the farmers. Seventy six percent of the farmers (377/499) reported <10 clinical cases of mastitis in the year prior to the questionnaire. Logistic multiple regression indicated that BMSCC >200 x 10(3)cells/ml was more likely when foremilking was practised, and when cows were collected in a yard before milking. BMSCC was less likely to be >200 x 10(3)cells/ml when teats were washed with water containing disinfectant compared with plain water; when the udder and teats were always checked before milking compared with, sometimes or never; when cows with mastitis were milked first compared with any other ordering, and when farmers recorded individual-cow somatic-cell count (ICSCC) compared with when ICSCC was not recorded. PMID- 12628769 TI - Modeling alternative mitigation strategies for a hypothetical outbreak of foot and-mouth disease in the United States. AB - Alternative mitigation strategies were compared during hypothetical outbreaks of foot-and-mouth disease (FMD) in the USA using a computer-simulation model. The epidemiologic and economic consequences were compared during these simulated outbreaks. Three vaccination and four slaughter strategies were studied along with two speeds of FMD virus spread among three susceptible populations of animals. The populations represented typical animal demographics in the United States. The best strategy depended on the speed of spread of FMD virus and the demographics of the susceptible population. Slaughter of herds in contact with known contagious herds was less costly than slaughtering only contagious herds. Slaughtering in 3 km rings around contagious herds was consistently more costly than other slaughter strategies. Ring vaccination in 10 km rings was judged more costly than slaughter alone in most situations. Although early ring vaccination resulted in lower government costs and duration in fast-spread scenarios, it was more costly when vaccinated animals were slaughtered with indemnity and other related slaughter costs. PMID- 12628770 TI - Morbidity of purebred dogs in Denmark. AB - Prevalences of diseases in purebred dogs are reported from a questionnaire study among members of the Danish Kennel Club (DKC) in 1997. The 4295 dogs in the study represented 24 breeds and 16 breed groups. Sixty percent of the dogs had at least one disease-event during their lifetime with a median of 2 and a maximum of 11. Diseases of the skin were the most-frequent (13.6%) followed by eye diseases (13.2%), accidents (12.6%) and diseases of the ear (12.6%). The prevalences of five diseases differed significantly between the breeds (diseases of the skin, eyes, ears, respiratory and skeletal systems). Males had significantly higher prevalences of respiratory diseases, skeletal diseases and diseases of the mouth and teeth. Tumours and diseases of the urinary system were most prevalent in females. Gender- and breed-specific prevalences are reported for the most frequent diseases. PMID- 12628771 TI - Mortality of purebred and mixed-breed dogs in Denmark. AB - Causes of death and age at death of 2928 dogs are reported from a questionnaire study among members of the Danish Kennel Club (DKC) in 1997. The dogs represented 20 breeds, 15 breed-groups and a group of mixed-breed dogs. The median age at death for all dogs in the study was 10.0 years. Mixed-breed dogs had a higher median age at death (11.0 years) than the entire population, but breeds like Shetland Sheepdog, Poodle and Dachshund exceeded this age (12 years). The Bernese Mountaindog, the group of Molossian types and the Sighthounds had the shortest life span with a median age at death of 7.0 years. Old age was the most frequent reported cause of death (20.8%) followed by cancer (14.5%), behavioural problems (6.4%) accidents (6.1%), hip dysplasia (4.6%), heart diseases (4.6%) and spinal diseases (3.9%). Breed-specific proportional mortalities with 95% confidence limits are given for the six most prevalent specific causes of death. PMID- 12628772 TI - The association of exercise, diet and other factors with owner-perceived obesity in privately owned dogs from metropolitan Perth, WA. AB - Randomly selected dog-owning households (n=657) were interviewed to identify exercise, dietary and other factors associated with obesity in dogs. Dogs (n=860) were categorised by their owners as underweight, correct-weight or overweight and the weight of 386 dogs was also recorded. Twenty five percent of dogs were reported as overweight. In random-effects logistic regression, overweight dogs had higher odds of being neutered (OR=2.8), fed snacks (OR=1.5), fed once a day (OR=1.4) or living in single-dog households (OR=1.6). For each year of age, the odds of obesity increased (OR=1.1) and for each hour of weekly exercise the odds of obesity decreased (OR=0.9). PMID- 12628773 TI - Risk factors associated with interdog aggression and shooting phobias among purebred dogs in Denmark. AB - The prevalence of behaviour problems is reported from a questionnaire study among members of the Danish Kennel Club (DKC). In total, 4359 dog owners were included in the analyses. With logistic regression, we analysed four behaviour problems: dominance towards the owner, interdog dominance aggression, separation anxiety and shooting phobia. Compared to Labrador Retrievers, the following breeds and breed groups had higher odds of being reported to have interdog dominance aggression: Belgian Sheepdogs, Dachshunds, Dalmatians, German Shepherds, Hovawarts, Pinschers, Rottweilers, Scent dogs and Spitz dogs. Poodles, retrieving/flushing dogs, Sheepdogs, Spitz dogs and terriers had higher odds of shooting phobia. The odds of interdog dominance aggression were higher among dogs owned by younger dog owners compared to dogs owned by older dog owners. Dogs living in the capital area of Copenhagen had increased odds of interdog dominance aggression as compared to dogs living in other parts of Denmark. Dogs belonging to owners with limited knowledge of the breed before acquiring the dog had higher odds of interdog dominance aggression. Dogs attending obedience training classes had reduced odds of shooting phobia. Dogs belonging to dog breeders had reduced odds of being reported to have the investigated behaviour problems. PMID- 12628774 TI - Evaluation of the mass spectrometric patterns of volatile liquid anaesthetics to predict the temperature--metal caused decomposition pathway. AB - In this study, prediction of the thermal decomposition pathway of the volatile liquid anaesthetics such as halothane, enflurane and isoflurane in contact with various metal/metal oxides at elevated temperatures has been deduced by evaluating the mass spectrometric fragmentation pattern of each anaesthetic observed in the ionisation process. In the light of the molecular structures and fragmentation components, it was believed that the thermal decomposition pathway of each anaesthetic on metal/metal oxide surface, particularly at higher temperatures, is similar to the ionisation mechanism occurring in the mass spectrometer ionisation process with minor differences for each molecule. The ionisation clusters for each anaesthetic molecule show the most likely fragment and radicals formed in the mass spectrometric ionisation process by considering the isotopic effect. From these clusters, thermal decomposition pathway of the liquid anaesthetics and formation mechanisms of the major halogenated thermal decomposition products have been predicted. It was concluded that the ionisation and thermal decomposition pathway resembles each other, but are not completely similar. PMID- 12628775 TI - Prediction of reactive hazards based on molecular structure. AB - There is considerable interest in prediction of reactive hazards based on chemical structure. Calorimetric measurements to determine reactivity can be resource consuming, so computational methods to predict reactivity hazards present an attractive option. This paper reviews some of the commonly employed theoretical hazard evaluation techniques, including the oxygen-balance method, ASTM CHETAH, and calculated adiabatic reaction temperature (CART). It also discusses the development of a study table to correlate and predict calorimetric properties of pure compounds. Quantitative structure-property relationships (QSPR) based on quantum mechanical calculations can be employed to correlate calorimetrically measured onset temperatures, T(o), and energies of reaction, deltaH, with molecular properties. To test the feasibility of this approach, the QSPR technique is used to correlate differential scanning calorimeter (DSC) data, T(o) and -deltaH, with molecular properties for 19 nitro compounds. PMID- 12628776 TI - A review of classic Fenton's peroxidation as an advanced oxidation technique. AB - Hydrogen peroxide (H(2)O(2)) is a strong oxidant and its application in the treatment of various inorganic and organic pollutants is well established. Still H(2)O(2) alone is not effective for high concentrations of certain refractory contaminants because of low rates of reaction at reasonable H(2)O(2) concentrations. Improvements can be achieved by using transition metal salts (e.g. iron salts) or ozone and UV-light can activate H(2)O(2) to form hydroxyl radicals, which are strong oxidants. Oxidation processes utilising activation of H(2)O(2) by iron salts, classically referred to as Fenton's reagent is known to be very effective in the destruction of many hazardous organic pollutants in water. The first part of our paper presents a literature review of the various Fenton reagent reactions which constitute the overall kinetic scheme with all possible side reactions. It also summarises previous publications on the relationships between the dominant parameters (e.g. [H(2)O(2)], [Fe(2+)], . . .). The second part of our review discusses the possibility of improving sludge dewaterability using Fenton's reagent. PMID- 12628777 TI - A review of thermal sludge pre-treatment processes to improve dewaterability. AB - As a result of the wide application and utilization of the waste activated sludge process, excess sludge presents a serious disposal problem. Many efforts have been devoted to reduce the excess sludge by treatments such as digestion and dewatering. It has been known for many years that a thermal pre-treatment gives an improvement in the dewaterability of sludges. This paper provides a literature review concerning the optimum treatment conditions to obtain enhanced dewaterability and digestibility of sludge. The main commercial hydrolysis processes (Cambi, Porteous and Zimpro) are discussed. The literature findings concerning the optimum treatment conditions of thermal or thermochemical pre treatments are reviewed. The second part of this paper deals with the fundamentals of improving sludge dewatering. The influence of extracellular polymer (ECP) on settling and dewatering characteristics is discussed, together with the importance of cations and ECP-hydrophobicity in the flocculation and dewatering process. Finally, the effect on exocellular polymer, dewaterability, settleability and colloidal stability of activated sludge by treatment with sulfuric acid was studied. PMID- 12628778 TI - Recent advances in VOCs removal from water by pervaporation. AB - Pervaporation (PV) is a separation process in which minor components of a liquid mixture are preferentially transported by partial vaporization through a non porous permselective (selectively permeable) membrane. PV is an emerging technology in environment cleanup operations, especially in the removal of volatile organic compounds (VOCs) from industrial wastewaters or contaminated groundwaters. Current state of PV membrane development in VOC removal and improvement in process engineering, and better understanding of the interactions between VOCs and membrane materials are reviewed. Among PV process parameters documented here are process temperature, permeate pressure, feed concentration, and feed flow rate. The effects of these parameters on PV selectivity and permeation flux have been studied extensively and these studies have borne fruit in a better understanding of many aspects of PV processes. The challenge in implementing PV in practical operations lies in the further enhancement of membrane quality for specific VOCs as well as improved management and control of possible adverse hurdles coming from real systems. PMID- 12628779 TI - Pilot-scale peroxidation (H2O2) of sewage sludge. AB - Municipal and industrial wastewater treatment plants produce large amounts of sludge. This excess sludge is an inevitable drawback inherent to the waste activated sludge process. Both the reduction of the amount of sludge produced and improving its dewaterability are of paramount importance. Novel pre-treatment processes have been developed in order to improve sludge dewatering, handling and disposal. This paper discusses the oxidation process utilising the catalytic activation of H(2)O(2) by iron salts, referred to as Fenton's reagent. In previous work, the authors described the experimental laboratory results of H(2)O(2)-oxidation of thickened sludge. Based upon the optimum conditions obtained in these laboratory tests, pilot-scale experiments are conducted. Peroxidation under its optimum conditions, i.e. (i) through addition of 25 g H(2)O(2) kg(-1) DS (dry solids content), (ii) in the presence of 1.67 g Fe(2+) ions kg(-1) DS, (iii) at pH 3, and (iv) at ambient temperature and pressure, significantly reduces the amounts of sludge and improves the product quality: the amount DS per equivalent inhabitant per day (DS/IE.d) was reduced from 60 to 33.1 g DS/IE.d and the percentage DS of the sludge cake was 47%, which is high compared with the 20-25% achieved in a traditional sludge dewatering facility. An economic assessment for a wastewater treatment plant of 300,000 IE confirms the benefits. Considering the fixed and variable costs and the savings obtained when the sludge is incinerated after dewatering, a net saving of approx. 950,000 Euro per year or 140 Euro per ton DS can be expected. PMID- 12628780 TI - Breakthrough of methyethylketone and benzene vapors in activated carbon fiber beds. AB - The breakthrough of low concentration methyethylketone (MEK) and benzene vapors in beds packed with rayon-based activated carbon fiber (ACF) with different surface areas was investigated. The breakthrough characteristics depend on the properties of the ACF and the vapors, as well as on the adsorption conditions. The results of dynamic adsorption in an ACF bed were consistent with those of equilibrium adsorption by gravimetric methods. The breakthrough adsorption indicates that ACF, with an appropriate surface area, could be utilized in controlling volatile organic compounds (VOCs) in indoor air. PMID- 12628781 TI - Adsorption of ammonium dinitramide (ADN) from aqueous solutions. 1. Adsorption on powdered activated charcoal. AB - Investigations on the adsorption of ammonium dinitramide (NH(4)N(NO(2))(2)) (ADN) from aqueous solutions on powdered activated charcoal (PAC) were carried out in order to find out an effective and easier method of separating ADN from aqueous solutions. The effectiveness of PAC in the selective adsorption of ADN from aqueous solutions of ADN (ADN-F) and ADN in presence of sulfate (SO(4)(2-)) and nitrate (NO(3)(-)) ions (ADN-PS) was examined and compared using batch and column methods. The adsorption process follows both Langmuir and Freundlich adsorption isotherms and the isotherm parameters for the models were determined. The observed data favor the formation of monolayer adsorption. The adsorption capacities were found to be 63.3, 119, 105.3 and 82 mg of ADN per g of PAC for ADN-F (batch), ADN-PS (batch), ADN-F (column) and ADN-PS (column), respectively. Break-through curves for ADN-F and ADN-PS were obtained for the optimization of separation of ADN from aqueous solutions. Elution curves were generated for the desorption of ADN from PAC using hot water as eluent. PMID- 12628782 TI - Tracer studies for evaluation of in situ air sparging and in-well aeration system performance at a gasoline-contaminated site. AB - Field-scale tracer studies were conducted at a gasoline-contaminated site in order to evaluate the effectiveness of in situ air sparging (IAS) and in-well aeration (IWA) in controlling the movement of soil gas and groundwater in the subsurface. The field site was comprised of silty sand (SM) and silty clay (CL), underlain by a clay layer at approximately 7.6 m. Depth to groundwater ranged from 2.4 to 3 m. Soil permeability and the natural hydraulic gradient were both low. Helium was used to trace the movement of soil gas in the unsaturated zone during the IAS field study, and successfully confirmed short-circuit pathways for injected air and demonstrated the limited distribution of injected gases at this site. Fluorescein, bromide, and rhodamine were used to trace the movement of groundwater during the IWA system field study, and successfully documented the inability of the IWA system to recirculate enough groundwater to enhance subsurface dissolved oxygen levels or to remediate groundwater by air stripping at this site. The inability of the systems to remediate the site was likely due to site conditions which consist of low-permeability soils and decreasing permeability with depth. As a result, relatively impermeable layers exist at the depth of the IAS screen and the lower IWA screen. These site conditions are not conducive to successful performance of either remediation system. PMID- 12628783 TI - Effect of amphiphilic polyurethane nanoparticles on sorption-desorption of phenanthrene in aquifer material. AB - Micelle-like amphiphilic nano-sized polyurethane (APU) nanoparticles were synthesized via chemical cross-linking reaction of nano-aggregates of urethane acrylate nonionomer (UAN) chain and were tested for extraction efficiency of sorbed phenanthrene from aquifer material. Even though the solubilizing performance and interfacial activity of APU nanoparticles were inferior to that of Triton X-100, in the low concentration region, APU nanoparticles could effectively reduce phenanthrene sorption on the aquifer material and extracted sorbed phenanthrene from the aquifer material, whereas Triton X-100 could not extract sorbed phenanthrene and rather increased phenanthrene sorption onto the aquifer materials. At higher concentrations, APU nanoparticles and Triton X-100 had almost the same soil washing effectiveness. This interesting result is mainly due to a lower degree of sorption of APU nanoparticles onto the aquifer material. The sorption of APU nanoparticles onto aquifer sand is largely hindered by their chemically cross-linked nature, resulting in better soil-washing performance of APU nanoparticles than Triton X-100. PMID- 12628784 TI - Acute public health consequences associated with hazardous substances released during transit, 1993-2000. AB - Massive quantities of hazardous substances are transported each day throughout the United States. While most arrive safely at their destination, uncontrolled releases of substances in transit do occur and have the potential of causing acute public health consequences for those individuals at or near the release. Data from 16 state health departments participating in the Agency for Toxic Substances and Disease Registry's (ATSDR) Hazardous Substances Emergency Events Surveillance (HSEES) system were analyzed to determine the public health consequences that occurred from actual releases in transit. Of the 9392 transportation events analyzed, 9.1% resulted in 2008 victims, including 115 deaths. The population groups injured most often were employees and the general public. The most common injury sustained was respiratory irritation. Evacuations were ordered in 5.5% of events effecting at least 63,686 people. Human error and equipment failure were the most common factors leading to events. These findings underscore the importance of job safety training, community planning, and effective emergency response to prevent adverse public health consequences from occurring or lessen their effect on the public. PMID- 12628785 TI - Role of humic acid entrapped calcium alginate beads in removal of heavy metals. AB - Most of the available techniques for removal of heavy metals from solution such as leachates from waste sites are very expensive. The technique described in this paper is designed to study the removal of humic acid by alginate beads and the subsequent binding of the metals within the beads thus decreasing the leachability of heavy metals from a hazardous waste site. The entry of 500 mg/l humic acid into 3% calcium alginate beads in 24 h showed a distinct fluorescence as compared to the fluorescence shown by the entry of 100 and 250 mg/l of humic acid into alginate beads. Further, on treatment of calcium alginate beads containing 500 mg/l humic acid with metals such as Cu, Mn, Zn, Cr and Fe, quenching of fluorescence was noticed, which was maximum with Cu and minimum with Fe. PMID- 12628786 TI - Effectiveness of cement-based systems for stabilization and solidification of spent pot liner inorganic fraction. AB - Approximately 7000 t of spent pot liner (SPL) wastes are generated annually from activities associated with Alumi;nio Brasileiro S.A. (ALBRAS) plant located at Barcarena, Para state, Brazil. The inorganic fraction of SPL contains high level of toxic compounds like cyanide and fluoride; its safe disposal has been the subject of serious discussions in Brazil. This study evaluated the option of a cement-based stabilization/solidification system as an effective means for safe disposal of SPL inorganic fraction in the field. The studies were carried out with concrete hexagonal blocks manufactured with a constant mass of 10% (w/w) of waste, 20% (w/w) of cement, and varied percentages of water, coarse aggregate, sand, and additives. The concrete matrices porosity and compressive strength were controlled by using microsilica (MS) and superplaticizer (SP). The results showed an average pH values for the SPL inorganic fraction and fragmented blocks of 10.2 and 11.1, respectively. Mixing the waste with concrete ingredients the solidification/stabilization effectiveness for the leachable cyanides and fluorides were of 59.33 and 57.95%, respectively. The results showed that the water/cement (W/C) ratio reduction through superplasticizer addition improved the compressive strength and the required value of 35 MPa was reached with blocks manufactured with 10 and 15% (weight of cement) of microsilica, after 28 days of curing time. PMID- 12628787 TI - Detoxification of methyl-parathion pesticide in aqueous solutions by electrochemical oxidation. AB - Commercial methyl-parathion (MeP) was detoxified using an electrochemical method that employed a Ti/Pt anode and stainless steel 304 as cathode. Sodium chloride was added as electrolyte and the mixture was passed through an electrolytic cell for 2 h. Due to the strong oxidizing potential of the produced chemicals, the organic pollutants were wet oxidized to carbon dioxide and water. A number of experiments were run at laboratory scale. Reductions of COD and BOD(5) were both over 80% and the mean energy consumption was 18-8 kWh per kg(-1) COD reduced (COD(r)). The degradation of MeP was more effective when the pH of the brine solution was in the acid range than when it was in the alkaline range. From the results it can be concluded that electrolysis could be used as an oxidation pre treatment stage for detoxification of toxic wastes with MeP. PMID- 12628788 TI - Photo-assisted oxidation of an oily wastewater using hydrogen peroxide. AB - The primary objective was to study the purification of an oily wastewater from a lubricant production unit using ultraviolet irradiation and hydrogen peroxide. The influence of hydrogen peroxide concentration, initial pH of the solution and of the addition of ferric ions on the chemical oxygen demand (COD) was examined. In each case, the concentration of the compounds contained in the oily wastewater was determined. It was shown that a 20-45% COD removal was achieved with 830-1660 mg l(-1) H(2)O(2). Gas chromatography-mass spectrometry analysis showed that the organic compounds of the wastewater decomposed to organic acids that were very resistant to photo-oxidation. Among these compounds, ethylene glycol remained almost unchanged by the attack from hydroxyl radicals. Acidic pH and Fe(III) addition enhanced significantly the photo-oxidation of the wastewater. PMID- 12628789 TI - Adsorption of reactive dyes on calcined alunite from aqueous solutions. AB - An attempt to alleviate the problem caused by the presence of reactive dyes in textile effluents was undertaken. Since alunite is a very abundant and inexpensive, we decided to experiment with it as a potential adsorbent for a certain type of the supracited pollutants used in cellulose fibers dyeing. The adsorption of Reactive Blue 114 (RB114), Reactive Yellow 64 (RY64) and Reactive Red 124 (RR124) by calcined alunite was studied by varying parameters such as the calcination temperature and time, particle size, pH, agitation time and dye concentration. Acidic pH was favorable for the adsorption of RB114 and alkaline pH was favorable to both RY64 and RR124. The equilibrium data fit the Langmuir isotherm. The adsorption capacities were found to be 170.7, 236 and 153 mg dye per gram of calcined alunite for RB114, RY64 and RR124, respectively. The pseudo first- and second-order kinetic models were used to describe the kinetic data, and the rate constants were evaluated. The experimental data were fitted by the second-order kinetic model, which indicates that chemicalsorption is the rate limiting step, inside of mass transfer. PMID- 12628790 TI - Disposal of MSWI fly ash through a combined washing-immobilisation process. AB - The objective of this work was to investigate the feasibility of a combined washing-immobilisation process as a means of optimising the disposal of fly ash resulting from municipal solid waste incineration (MSWI) in cementitious matrices. Two different types of Italian MSWI fly ash and an ordinary Portland cement (ASTM Type I) were used. Washing pre-treatment of fly ash with water always produces a wastewater that can be successfully treated by reducing the pH to values of 6.5-7.5. This treatment is capable of removing the detected contaminants (Al, Cd, Pb, Zn) through two different mechanisms: precipitation of aluminium hydroxide and adsorption of cadmium, lead and zinc ions onto floc particles of Al(OH)(3). Setting and leaching tests on cementitious mixes prove that the hazardous sludge produced from wastewater treatment can be completely mixed with washed fly ash and this mixture can be incorporated into cementitious matrices to a great extent (75 wt.% of total solid) without the risks of an unacceptable delay of cement setting and an excessive heavy metals leachability from solidified products. The better performance of the combined washing immobilisation process as compared to the immobilisation process of unwashed fly ash may be ascribed primarily to the ability of the washing step in promoting the formation of hydrate phases that incorporate and/or convert heavy metal compounds into less reactive forms and, secondarily, to its ability of removing significant amounts of alkali chlorides and sulphates from fly ash. As a result, MSWI fly ash is transformed into a material that adversely affects cement hydration to a much lower extent than unwashed fly ash. PMID- 12628791 TI - Oxidative degradation and detoxification of aqueous carbofuran by membrane anodic Fenton treatment. AB - Anodic Fenton treatment (AFT), a new Fenton technology for the treatment of pesticide wastewater, has been reported previously. The substitution of an ion exchange membrane for the salt-bridge, an improvement to the practicality of the AFT without sacrificing treatment efficiency, has also been reported. The oxidative degradation by membrane AFT of carbofuran, a heavily used and toxic carbamate insecticide, was investigated in this study. The results show that the degradation kinetics of carbofuran with different initial concentrations obeys the AFT model, and the treatment efficiency increases with increasing initial concentration. Raising the treatment temperature can result in enhanced degradation of carbofuran in solution. The pseudo-activation energy of carbofuran by membrane AFT was estimated to be 7.66 kJ mol(-1). The results also show that AFT treatment can effectively remove COD and dramatically improve the biodegradability of carbofuran in solution. GC/MS analysis found four degradation products, revealing that the carbamate branch and 3-C in the furan ring are the first and second attack targets of hydroxyl radicals. As shown by the toxicity assay, the fatal toxicity of carbofuran to earthworms can be totally removed. The degradation of carbofuran by AFT is also a detoxification process. PMID- 12628792 TI - Adsorptive removal of 2-chlorophenol by low-cost coir pith carbon. AB - Adsorption of 2-chlorophenol (2-CP) by coir pith carbon was carried out by varying the parameters such as agitation time, 2-CP concentration, adsorbent dose, pH and temperature. Adsorption equilibrium reached at 40, 60, 80 and 100 min for 2-CP concentration of 10, 20, 30 and 40 mg/l, respectively. Adsorption followed second-order kinetics. The adsorption equilibrium data obeyed Freundlich isotherm. Acidic pH was favorable for the adsorption of 2-CP. Desorption studies showed that chemisorption plays a major role in the adsorption process. PMID- 12628793 TI - Hot acid hydrolysis as a potential treatment of thickened sewage sludge. AB - Municipal and industrial wastewater treatment plants produce large amounts of sludge, containing organic and mineral components and being mechanically dewatered to e.g. 20-25% DS in centrifuges. Both the reduction of the amount of sludge produced and improving its dewaterability are hence of paramount importance. Hot acid hydrolysis can meet these objectives. The current paper describes the results of detailed investigations with respect to acid hydrolysis of thickened sludge (5-6% DS content). A comparison with traditional thermal hydrolysis is also included. As a result of the experimental investigations, it can be concluded that hot acid hydrolysis is efficient in both reducing the residual sludge amounts and improving the dewaterability. Under the proposed optimum conditions it is found that (i) the amount of hydrolysed DS is approximately 70% lower than the initial untreated amount, (ii) the DS-solid content of the dewatered cake is increased from 22.5% (initial untreated) to at least twice this value, (iii) the rate of mechanical dewatering is not significantly affected. The preferential release of ODS into the water phase, and the increased BOD/COD-ratio through hydrolysis, turn this recycle water phase into a possible carbon-source for nitrification/denitrification. Heavy metals and phosphates are also released in the water phase, and can be subsequently precipitated. PMID- 12628794 TI - Industrial wastewater treatment in a new gas-induced ozone reactor. AB - The present work was to investigate industrial wastewater treatment by ozonation in a new gas-induced reactor in conjunction with chemical coagulation pretreatment. The reactor was specifically designed in a fashion that gas induction was created on the liquid surface by the high-speed action of an impeller turbine inside a draft tube to maximize the ozone gas utilization. A new design feature of the present reactor system was a fixed granular activated carbon (GAC) bed packed in a circular compartment between the reactor wall and the shaft tube. The fixed GAC bed provided additional adsorption and catalytic degradation of organic pollutants. Combination of the fixed GAC bed and ozonation results in enhanced oxidation of organic pollutants. In addition to enhanced pollutant oxidation, ozonation was found to provide in situ GAC regeneration that was considered crucial in the present reaction system. Kinetic investigations were also made using a proposed complex kinetic model to elucidate the possible oxidation reaction mechanisms of the present gas-induced ozonation system. As a complementary measure, chemical coagulation pretreatment was found able to achieve up to 50% COD and 85% ADMI removal. Experimental tests were conducted to identify its optimum operating conditions. PMID- 12628795 TI - UV-light induced photodegradation of 17alpha-ethynylestradiol in aqueous solutions. AB - The photodegradation of 17alpha-ethynylestradiol (EE2) in aqueous solutions induced by UV-light was preliminarily studied in this paper by means of fluorescence, UV and infrared spectra. The result suggested that EE2 in aqueous solutions underwent photodegradation under irradiation with UV disinfection lamp (lambda = 254 nm, 30 W), but the photodegradation was not observed under high pressure mercury lamp (lambda > or = 365 nm, 250 W). The photodegradation of 1.6 20.0 mg/l EE2 in aqueous solutions at a given initial pH value of 6.8 was pseudo first order reaction. Increasing the initial concentration of EE2 lowered the photodegradation rate. The photodegradation rate of EE2 reached the lowest value at pH about 5.0, higher pH values of 6.0-8.0 benefited the photodegradation. Ferric ions can promote the photodegradation of EE2 in aqueous solutions at pH value of 2.0-5.0. PMID- 12628808 TI - Tick-borne encephalitis. PMID- 12628809 TI - Molecular aspects of TBE virus research. PMID- 12628810 TI - Tick-borne encephalitis--pathogenesis, clinical course and long-term follow-up. AB - The prospective studies available today confirm the experience gained from several retrospective studies that TBE is a disease with a severe acute clinical course and considerable long-term morbidity. A defined post-encephalitic TBE syndrome exists, causing long-lasting morbidity that often affects the quality of life and sometimes also forces the individual to a change in life-style. The sequelae render high costs for individual patients and the society. Three clinical courses may be identified: one with complete recovery within 2 months, occurring in approximately one fourth of patients, one with protracted, mainly cognitive dysfunction, and one with persisting spinal nerve paralysis with or without other post-encephalitic symptoms. Up to 46% of patients are left with permanent sequelae at long-time follow-up, the most commonly reported residuals being various cognitive or neuropsychiatric complaints, balance disorders, headache, dysphasia, hearing defects, and spinal paralysis. This knowledge enhances the need for continued local epidemiological surveillance of TBE to form a basis for vaccination policies. Even though knowledge of the clinical course of TBE has improved in recent years, there are still several aspects of this disease that warrant further studies. These comprise the clinical picture and prognosis in children, an evaluation of different rehabilitation strategies, and an improved understanding of pathogenic mechanisms to permit the development of antiviral or, maybe more probable, immune modulatory treatment strategies. PMID- 12628811 TI - Epidemiology and ecology of TBE relevant to the production of effective vaccines. AB - This review focuses on the epidemiology and ecology of the tick-borne encephalitis (TBE) virus including all the factors influencing the ecology of the TBE virus, environmental and climatic conditions, the vectors and reservoir hosts and their interactions. Consequently, the structure and the conditions of natural foci of TBE are described, as well. Special emphasis is given on data regarding the virus prevalence in ticks in the field. In the second part of this review all the epidemiological and surveillance data of TBE in the 27 European countries and in the Far East with risk areas of TBE and in China and Japan are summarised. Therefore the review is a basis for travellers and inhabitants to get background information for their personal risk assessment. PMID- 12628812 TI - Diagnosis of tick-borne encephalitis. AB - The actual diagnosis of a tick-borne encephalitis (TBE) must be established in the laboratory because of the non-specific clinical features it presents. The method of choice is the demonstration of specific IgM- and IgG-serum antibodies by enzyme-linked immuno-sorbent assay (ELISA), since these antibodies are detectable in practically every case at the time of hospitalization. Early after onset of disease in the cerebrospinal fluid specific antibodies can only be found in 50% of the patients, but by the 10th day of illness they almost invariably become detectable. If an infection occurs after and despite the post-exposure administration of a specific immunoglobulin the seroconversion can be delayed and may cause diagnostic problems. Virusisolation from the blood, or the detection of specific nucleic acid in the blood or the cerebrospinal fluid by reverse transcriptase polymerase chain reaction (RT-PCR) usually is only successful during the first viremic phase of the disease before seroconversion. In fatal cases, the virus can be isolated or detected by RT-PCR from the brain and other organs. For testing immunity after a TBE virus infection or after vaccination, most often the IgG ELISA is used. However, in cases of other flavivirus contacts (e.g. vaccinations against yellow fever or Japanese encephalitis; dengue virus infections), the performance of a neutralization assay is necessary for assessing immunity due to the interference of flavivirus cross-reactive antibodies in ELISA and hemagglutination inhibition (HI) test. PMID- 12628814 TI - TBE vaccination and the Austrian experience. AB - In the pre-vaccination era, Austria had the highest recorded morbidity of tick borne encephalitis (TBE) in Europe. The disease accounted for more than 50% of all viral meningoencephalitides in the eastern and southern parts of the country. This prompted us to start a cooperative vaccine development project in 1971 with J. Keppie from the Microbiological Research Establishment, Porton Down, England. After very satisfactory results of field studies, conducted in individuals for whom TBE was classified as occupational disease (forest workers, farmers, etc.), the killed virus vaccine (see chapter by N. Barrett in this issue), was made commercially available by Immuno AG Vienna (now Baxter Health-Care). The vaccine proved to be highly immunogenic and very well tolerated in both adults and children. After completing the series of three vaccinations seroconversion rates of >99% were recorded. In Austria, as in other European countries, TBE is now acquired for the most part during leisure activities. In view of this fact a mass vaccination campaign was initiated in 1981. Subsequently, the vaccination coverage of the Austrian population increased from 6% in 1980 to 86% in 2001, exceeding 90% in some of the high-risk areas. Data annually collected by our surveillance system show that the clinical effectiveness of the vaccine is excellent. Based on the assumption that the whole Austrian population is at risk of infection the calculated rate of protection after three doses of the vaccine is 96-98.7%. Breakthrough disease is rare and affects mainly higher age groups. Since the advent of TBE vaccination only one single case has been observed in the age group up to 20. The increasing vaccination coverage led to a more or less steady decline of TBE, drastically reducing the public health problem that the disease poses in Austria, especially in the provinces, where formerly the highest morbidity rates were observed. For example, in Carinthia, in the years 1973-1982 an average annual incidence of 155 was recorded, compared with only four annual cases in the last 4 years. The Austrian experience shows that containment of a tick-borne viral disease is feasible, provided a well-tolerated and effective vaccine is available that is widely accepted by the general population. PMID- 12628813 TI - History of TBE vaccines. AB - Tick borne encephalitis was a frequent viral CNS disease in Europe in the last century, with several hundred hospitalisation cases per year. The causative agent is tick-borne encephalitis virus (TBEV), a flavivirus that is transmitted by ticks. The first tissue culture derived vaccine was developed in the early 1970s and this was commercialised as a collaboration between the Institute of Virology, Vienna, Austria and Immuno AG Vienna (now Baxter Healthcare). This vaccine was highly successful during the last thirty years and its use resulted in a significant reduction in the incidence of disease in central Europe, particularly Austria. A number of modifications have been made to the manufacturing process and formulation of this vaccine over this period and a second manufacturer has also entered the market. This article describes the development and modification of these vaccines over this time frame, and illustrates the efficacy of the vaccine in preventing this severe CNS disease. PMID- 12628816 TI - Social marketing and the establishment of the ISW-TBE. AB - Vaccination can only be offered effectively to the general public by the means of social marketing. Experience gained with tick-borne encephalitis (TBE) in Austria will be used to demonstrate the need and importance of this public health tool. After the realisation that only mass vaccination would effect a decrease in the number of TBE cases, a nation-wide TBE vaccination programme was initiated in 1981. Since then there has been a dramatic decrease in the incidence of TBE. The annual TBE-vaccination campaign is by far the most visible of all public health programmes in Austria and has been instrumental in reaching the present vaccination rate of 86% in this country. Outside of Austria TBE is rapidly becoming a growing public health problem in Europe and other parts of the world. In order to effectively contribute toward public health in general and, in specific, to encourage the control of TBE, an international effort was launched with the aim of alleviating this situation. As a first step towards this objective, renowned international experts on TBE created a new body: The International Scientific Working Group on Tick-Borne Encephalitis (ISW-TBE). This Working Group is comprised of internationally recognised scientific experts from endemic and non-endemic regions with extensive personal expertise in the field of TBE and a high level of commitment to improving the knowledge of and response to TBE. PMID- 12628815 TI - A controlled trial of tick-borne encephalitis vaccination in patients with multiple sclerosis. AB - The aim of this study was to investigate a possible link between a single vaccination against tick-borne encephalitis (TBE) and the appearance of one or more new cerebral lesions in magnetic resonance imaging (MRI) and/or a clinical relapse of MS. Fifteen MS patients with documented history of MS relapses living in a TBE endemic area were matched with 15 patients selected from a patient database containing 500 cases of MS. Three patients in each group were unvaccinated while all others had basic immunisation and regular booster vaccinations. Patients of the vaccination group received a single dose (3.3 microg) of a TBE vaccine. TBE antibodies were detected by ELISA and confirmed by neutralisation test. MRI was used as marker for disease activity and progression in addition to the clinical neurological examination. No association was seen between TBE vaccination and MRI detected disease activity, clinical relapse or disease progression of MS. PMID- 12628817 TI - Report of the Meningitis Program of the International Scientific Working Group on TBE. Serological screening of patients with viral CNS-infection of unknown etiology in search of undiagnosed TBE cases. AB - The endemicity of tick-borne encephalitis (TBE) in Europe is changing. Potential undetected or emerging TBEV foci and the risk of underdiagnosis due to a low awareness among the medical community form the background of this retrospective multicenter follow-up study. We investigated the possibility of undiagnosed TBE cases among patients with presumed viral central nervous system (CNS)-infection of unknown etiology. Eight centers in four European countries provided sera and/or cerebrospinal fluid (CSF) samples from 233 individuals. The samples were screened with a commercial TBEV ELISA test system (IgM and IgG). Positive or borderline samples were re-evaluated at the Institute of Virology in Vienna by an in-house ELISA test and a neutralization test (NT). Two previously undiagnosed Swedish TBE patients were verified. Three additional individuals from Swedish centers were IgG ELISA and NT positive. No NT positive individuals were found from France, Belgium or The Netherlands. Nineteen individuals were found IgG TBE ELISA positive, but negative in NT, indicating unspecific reactivity. At least four of those patients were vaccinated against yellow fever. The probable reason for the reactivity seen in these individuals is the well-known cross-reactivity existing among flaviviruses. PMID- 12628818 TI - The Austrian TBE Patient Support Group. PMID- 12628819 TI - Enzyme induced biodegradation of polycarbonate-polyurethanes: dose dependence effect of cholesterol esterase. AB - The current study has investigated the influence of esterase activity (80 400units/ml) on the biodegradation of polycarbonate-urethanes (PCNUs) by cholesterol esterase (CE), with a particular interest in studying the influence of different hard segment structures and their contribution to sensitizing the polymer towards enzyme catalyzed hydrolysis. Polycarbonate based polyurethanes were synthesized with varying hard segment content as well as hard segment chemistry based on three different diisocyanates, 1,6-hexane diisocyanate (HDI), 4,4'-methylene bisphenyl diisocyanate (MDI) and 4,4-methylene biscyclohexyl diisocyanate (HMDI). The effect of different chemistry on surface contact angle was measured in order to define the relative chemical nature of the surfaces. The enzyme dose response was found to be lower when hard segment content in the polymer was high. There was a very strong dependence on enzyme concentration for polyurethanes with different hard segment chemistry, despite the fact that the nature of the hydrolysable polycarbonate segment remained the same. The PCNU which showed the most dramatic dependence on enzyme concentration was synthesized with HMDI. At low enzyme concentration (80units/ml) this material was the most stable of the polymers while at elevated CE concentration (400units/ml) the polymer underwent a catastrophic breakdown. The findings suggested that protein binding on the surfaces was saturated even though enzyme degradation did not achieve saturation on any of the surfaces. The role of protein binding in modulating the hydrolytic action of the enzymes at different activity levels highlights a need for further study in this area. PMID- 12628820 TI - Effects of transforming growth factor-beta 1 (TGF-beta1) on in vitro mineralization of human osteoblasts on implant materials. AB - An in vitro cell-implant mineralization model system was used to study the effect of transforming growth factor-beta 1(TGF-beta1) on mineralization in human osteoblast cultures. SaOs-2 and primary human osteoblast (HOB) cells were cultured on Tivanium (Ti-6Al-4V) disks. Administration of different concentrations of TGF-beta1 (0.02, 0.01, 0.2, 1.0, 2.0ng/ml) to these cultures demonstrated a biphasic dose response with 0.2ng/ml TGF-beta1 maximally increasing the calcium content compared to control culture. Results with SaOs-2 and HOB cultures were similar. An optimal dose of TGF-beta1 (0.2ng/ml) was provided to the cultures either in one single dose or multiple doses. Continuous administration of 0.2ng/ml TGF-beta1 caused 77% (SaOs-2) and 60% (HOB) increases in calcification compared to the control and 0.2ng/ml single dose groups. Single administration of the accumulative dose at 1.6ng/ml had no significant effect on the calcium content in either cell culture compared to control. Two weeks continuous administration of 0.2ng/ml TGF-beta1 in both cell cultures resulted in significant increases in the expression of bone specific extracellular matrix proteins which included alkaline phosphatase, Type I collagen, and osteocalcin as measured by Northern blot analysis and RT-PCR. At 4 weeks, the mRNA level of Type I collagen was still significantly higher in the TGF-beta1 treatment group compared to control. In conclusion, TGF-beta1 enhances mineralization of HOB on implant materials. PMID- 12628821 TI - The use of trans-vinylene formation in quantifying the spatial distribution of electron beam penetration in polyethylene. Single-sided, double-sided and shielded irradiation. AB - The use of the quantification of trans-vinylene unsaturations in irradiated ultrahigh molecular weight polyethylene (UHMWPE) in determining the spatial distribution of electron beam penetration was investigated. UHMWPE was irradiated with a 10MeV linear electron beam accelerator to 100kGy at both room temperature and 125 degrees C in air. The irradiation was carried out in shielded and unshielded single-sided and unshielded double-sided irradiation modes. Dose-depth profiles were measured by quantifying the yields of trans-vinylene unsaturations as a function of depth away from the e-beam incidence surfaces using infrared spectroscopy. The extent of beam penetration increased with increasing irradiation temperature as was manifested by the increase in the iso-dose penetration with the unshielded single-sided irradiation and increased dose overlap with the unshielded double-sided irradiation. The optimum thickness with maximum uniformity in dose-depth distribution for double-sided irradiation was 85 and 90mm for 25 degrees C and 125 degrees C irradiation temperatures, respectively. PMID- 12628822 TI - The chitosan prepared from crab tendon I: the characterization and the mechanical properties. AB - Crystalline chitosan was prepared from crab tendon consisting mainly of chitin, including various proteins and calcium phosphates. The crab tendon has high mechanical properties due to its aligned molecular structure. Crab tendon components, i.e. proteins and calcium phosphates, were removed by deacetyl treatment using 50wt% NaOH aqueous solution at 100 degrees C, and a subsequent ethanol treatment. As judged from microscopic observations using an optical polarizer, the treated chitosan remained intact regarding its aligned molecular structure, and had a high tensile strength of 67.9+/-11.4MPa. The tensile strength was further enhanced to 235+/-30MPa by a thermal treatment at 120 degrees C, corresponding to the formation of the intermolecular hydrogen bonds. PMID- 12628823 TI - Carbonated calcium phosphates are suitable pH-stabilising fillers for biodegradable polyesters. AB - Carbonated amorphous calcium phosphates were prepared with different carbonate content. Their ability to neutralise acidity was probed by time-resolved titration experiments with lactic acid, the monomer that results from degradation of polylactide. The results show that although calcium phosphate as such can reduce acidity, their buffering range lies at a pH of about 4, i.e. outside the physiological range. This is not related to the rate of dissolution. Carbonated calcium phosphates as well as calcium carbonate (calcite) alone are able to keep the pH around 7.4. Consequently, carbonated calcium phosphates are suitable basic filler materials as they are able to compensate acidity, and to buffer within the physiological pH-range. PMID- 12628824 TI - Fabrication of a protein film by electrospray deposition method and investigation of photochemical properties by persistent spectral hole burning. AB - Transparent protein film of iron-free cytochrome c (Cyt. c) was successfully manufactured by electrospray deposition (ES) method and the properties of the protein film were investigated. Excursion temperature dependence of spectral hole profiles in the photochemical hole burning for the iron-free Cyt. c in protein film and in glassy solution was investigated and compared with that of iron-free porphyrin embedded in a synthetic polymer film to clarify photochemical properties of electrospray deposited protein film. The spectral holes of iron free porphyrin were thermally more stable in Cyt. c than in polymer, indicating compact packing of the chromophore in the protein. The ES-deposited iron-free Cyt. c in protein film showed less stable spectral hole than that in glassy solution. This difference is attributable to the electrospray deposition process of the protein involving ionization and to subsequent cross-linking of protein molecules. PMID- 12628825 TI - New micelle-like polymer aggregates made from PEI-PLGA diblock copolymers: micellar characteristics and cellular uptake. AB - New amphiphilic block copolymers based on oligomeric polyethylenimine and poly(D,L-lactide-co-glycolide) (PEI-PLGA) were synthesized by directly coupling PLGA with a carboxyl terminal group to PEI. The block copolymers were prepared by varying the length of the hydrophobic PLGA block (M(n)=6, 10, and 21K), while that of the hydrophilic PEI block (M(n)=423) was fixed. PEI-PLGA block copolymers were found to be self-assembled in water by using a PLGA segment as a hydrophobic aggregate block and a PEI segment as a hydrophilic corona-forming block. The block copolymers formed micelle-like aggregates with critical association concentration (cac) in the range of 1.54-2.57x10(-3)g/l in water. It was found that the size and cac of the aggregates depended on the hydrophobic block length and the ionic state of the PEI block. The aggregate size decreased and the cac increased, when the PLGA block length decreased and the PEI block was protonated. As a general program aimed at the development of a new nanoscopic drug carrier, the cellular uptake behavior of PEI-PLGA aggregates was compared with that of plain PLGA nanoparticles by using confocal microscopy. The results showed that PEI-PLGA aggregates was readily adsorbed onto the cell surfaces and translocated into the cytoplasm, implying their versatile applicability as a drug carrier. PMID- 12628826 TI - High resolution transmission electron microscopy of age-hardenable Au-Cu-Zn alloys for dental applications. AB - Microstructures of age-hardenable AuCu-Zn pseudobinary alloys for dental applications were studied by means of high resolution transmission electron microscopic (HRTEM) observation and X-ray diffraction study. HRTEM study revealed that the appearance frequency of antiphase boundaries (APBs) per unit volume of the AuCu II superstructure effectively increased by Zn addition to AuCu, which may be the reason for that high hardness was maintained for a long time in AuCu Zn alloys. The disordered APBs zone in the AuCu II superstructure had wavy characteristics and fluctuated within regular range. With increasing Zn content in AuCu-Zn alloys, the fluctuation range of APBs' width became narrower, thus random APBs' spacing and irregular APBs' shape of AuCu II superstructure changed to comparatively regular APBs' spacing and shape. Due to the APBs' wavy characteristics, spacing between successive APBs, M, was not constant but scattered, and the magnitude of the scattering of M value decreased with increasing Zn content. By Zn addition to AuCu, phase transformation from a disordered alpha phase to AuCu II phase was greatly accelerated, which made it possible for the AuCu-Zn alloy to have excellent age-hardenability at relatively low temperature like intraoral temperature. PMID- 12628827 TI - Albumin and fibrinogen adsorption on PU-PHEMA surfaces. AB - Materials that adsorb specific proteins may find a variety of applications in the biomedical field. The aim of this study was the preparation of a hydrophilic surface, with low protein adsorption, to be used in the future as a support for the immobilisation of several species, e.g. Cibacron Blue F3G-A, which has been described to induce specific albumin adsorption. Poly(hydroxyethylmethacrylate) (PHEMA) and poly(hydroxyethylacrylate) (PHEA) were chosen as the hydrophilic surface because they can be easily polymerised and possess hydroxyl groups that can be used for the immobilisation of different compounds. Thin films of PHEMA and PHEA were successfully graft polymerised onto the surface of a commercial poly(etherurethane) (PU) using ceric ion as initiator. Grafting polymerisations were followed by mass gain and attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR). Since stability tests demonstrated that only PU PHEMA was stable in alkaline solutions, a necessary condition to future immobilisations, the investigation was focused on the coating of PU with PHEMA. PU-PHEMA films were characterised in detail using several techniques as mass gain, ATR-FTIR, contact angle measurements, X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM). Protein adsorption was evaluated using radiolabelled albumin and fibrinogen from pure solutions and from mixtures of both proteins. PU surfaces modified with PHEMA have demonstrated low protein adsorption, showing their potential use as substrates. This opens the possibly of exploring the advantages of selective adsorption by appropriate immobilisation of specific molecules. PMID- 12628828 TI - A biodegradable nanofiber scaffold by electrospinning and its potential for bone tissue engineering. AB - Microporous, non-woven poly( epsilon -caprolactone) (PCL) scaffolds were made by electrostatic fiber spinning. In this process, polymer fibers with diameters down to the nanometer range, or nanofibers, are formed by subjecting a fluid jet to a high electric field. Mesenchymal stem cells (MSCs) derived from the bone marrow of neonatal rats were cultured, expanded and seeded on electrospun PCL scaffolds. The cell-polymer constructs were cultured with osteogenic supplements under dynamic culture conditions for up to 4 weeks. The cell-polymer constructs maintained the size and shape of the original scaffolds. Scanning electron microscopy (SEM), histological and immunohistochemical examinations were performed. Penetration of cells and abundant extracellular matrix were observed in the cell-polymer constructs after 1 week. SEM showed that the surfaces of the cell-polymer constructs were covered with cell multilayers at 4 weeks. In addition, mineralization and type I collagen were observed at 4 weeks. This suggests that electrospun PCL is a potential candidate scaffold for bone tissue engineering. PMID- 12628829 TI - Fabrication of homogeneously cross-linked, functional alginate microcapsules validated by NMR-, CLSM- and AFM-imaging. AB - Cross-linked alginate microcapsules of sufficient mechanical strength can immunoisolate cells for the long-term treatment of hormone and other deficiency diseases in human beings. However, gelation of alginate by external Ba(2+) (or other divalent cations) produces non-homogeneous cross-linking of the polymeric mannuronic (M) and guluronic (G) acid chains. The stability of such microcapsules is rather limited. Here, we show that homogeneous cross-linking can be achieved by injecting BaCl(2) crystals into alginate droplets before they come into contact with external BaCl(2). The high effectiveness of this crystal gun method is demonstrated by confocal laser scanning microscopy and by advanced nuclear magnetic resonance imaging. Both techniques gave clear-cut evidence that homogeneous cross-linkage throughout the microcapsule is only obtained with simultaneous internal and external gelation. Atomic force microscopy showed a very smooth surface topography for microcapsules made by the crystal gun method, provided that excess Ba(2+) ions were removed immediately after gelation. In vitro experiments showed greatly suppressed swelling for crystal gun microcapsules. Even alginate extracted from Lessonia nigrescens (highly biocompatible) yielded microcapsules with long-term mechanical stability not hitherto possible. Encapsulation of rat islets, human monoclonal antibodies secreting hybridoma cells and murine mesenchymal stem cells transfected with cDNA encoding for bone morphogenetic protein (BMP-4) revealed that injection of BaCl(2) crystals has no adverse side effects on cell viability and function. However, the release of low-molecular weight factors (such as insulin) may be delayed when using alginate concentrations in the usual range. PMID- 12628830 TI - ESR study of camphorquinone/amine photoinitiator systems using blue light emitting diodes. AB - New light-activation units equipped with high-illuminant blue light-emitting diodes (LEDs) have recently been proposed as a replacement for the halogen units that are widely used in dentistry to polymerize light-cured resins. The photoinitiators in light-cured dental resins, typified by the camphorquinone (CQ)/amine photoinitiator system, generate primary radicals with light irradiation that attack the double bonds of resin monomers. The physical properties of the cured resins are affected by the generation of primary radicals during the initial stage of polymerization. This study examined two types of photoinitiator systems, CQ/DMPT and CQ/DMAEMA, and three types of curing units, a new LED unit and two conventional halogen units. The primary radicals generated by irradiation were quantified using electron spin resonance (ESR) spectroscopy with a trapping method, using phenyl-tert-butyl nitrone as the trapping agent. The energy efficiencies of the LED and halogen units were compared by quantifying the generated radicals and emitted light energy (J/cm(2)). The energy required to generate a given amount of radicals using the LED unit was smaller than that using the halogen units (p<0.05). These results suggest that the new LED unit performs better than conventional halogen units with respect to light energy. PMID- 12628831 TI - Collapse pressures of biodegradable stents. AB - Biodegradable stent prototypes were produced from poly L-lactic acid polymers with different molecular weights. The effects of molecular weight, drug incorporation and stent design on the collapse pressure of the stents were evaluated. While molecular weights did not show a significant effect on the collapse pressure of the stents, drug incorporation at high percentage decreased the collapse pressure of the stents substantially. Cryogenic fracture surfaces showed significant drug agglomeration as the concentration increased. The design of the stent was also found to a have significant effect on the collapse pressure. The stent produced from the same material has a higher collapse pressure when the load bearing surface area is increased. PMID- 12628832 TI - Delayed fracture of beta titanium orthodontic wire in fluoride aqueous solutions. AB - Hydrogen embrittlement of a beta titanium orthodontic wire has been examined by means of a delayed-fracture test in acid and neutral fluoride aqueous solutions and hydrogen thermal desorption analysis. The time to fracture increased with decreasing applied stress in 2.0% and 0.2% acidulated phosphate fluoride (APF) solutions. The fracture mode changed from ductile to brittle when the applied stress was lower than 500MPa in 2.0% APF solution. On the other hand, the delayed fracture did not occur within 1000h in neutral NaF solutions, although general corrosion was also observed similar to that in APF solutions. Hydrogen desorption of the delayed-fracture-tested specimens was observed with a peak at approximately 500 degrees C. The amount of absorbed hydrogen was 5000-6500 mass ppm under an applied stress in 2.0% APF solution for 24h. It is concluded that the immersion in fluoride solutions leads to the degradation of the mechanical properties and fracture of beta titanium alloy associated with hydrogen absorption. PMID- 12628833 TI - Dose-adjusting epirubicin in patients with altered liver function: when classical pharmacology makes good practical sense. PMID- 12628834 TI - Altered fractionation and combined radio-chemotherapy approaches: pioneering new opportunities in head and neck oncology. AB - Squamous cell carcinoma of the head and neck (HNSCC) are increasingly treated by multimodality approaches combining surgery, radiotherapy and chemotherapy. Randomised controlled trials have demonstrated major improvements in loco regional tumour control from altered fractionation radiotherapy, accelerated fractionation and hyperfractionation, as compared with conventional fractionation. This experience is summarised, and the limit as to how far these modifications can be taken is discussed. It is emphasised that radiation fractionation will need to be optimised separately in multimodality strategies. Combined chemotherapy and radiotherapy has also been shown in phase III trials to produce an improved survival and an improved disease control. Chemotherapy may be given as neoadjuvant, concurrent or adjuvant treatment and the biological rationales for each of these, and the data supporting them, are reviewed. Although, large meta-analyses have shown concurrent chemoradiation to be the most effective, there is still a strong rationale for trying to develop neoadjuvant and adjuvant schedules. New, more active drugs may be important in this context. As therapy is becoming more intense, a careful recording and reporting of treatment-related morbidity is a crucial element in estimating the therapeutic gain from competing therapeutic management strategies. Development of non cytostatic drugs and individualization of therapy using molecular prognostic markers are exciting areas of research with a great potential for improving therapy in the next decade and these are briefly discussed. Finally, a number of avenues for further research are identified. PMID- 12628835 TI - Anti-oestrogenic chemoprevention of breast cancer-the need to progress. AB - Clinical trials of the selective oestrogen receptor modulator (SERM), tamoxifen, have shown an early reduction in risk of breast cancer in healthy women of approximately 40%, but with associated risks and benefits to normal tissues. An overall clinical benefit and the identification of the women at risk of breast cancer who may gain benefit from tamoxifen has not been clearly established. The identification of those women at risk who are most likely to gain benefit, and the development of other SERMs and aromatase inhibitors which might be more active and have a more beneficial spectrum of activity on normal tissues in healthy women is essential, if the aim of preventing breast cancer in healthy women is to be achieved. PMID- 12628836 TI - Epirubicin in patients with liver dysfunction: development and evaluation of a novel dose modification scheme. AB - This study aimed to develop an epirubicin dose modification scheme in women with breast cancer and liver dysfunction. We first identified target areas under the concentration-time curve (AUCs) of 2400 and 1600 ng/ml.h from pharmacokinetic studies in 15 women with normal liver tests. In a second group of 16 women with abnormal liver biochemistry, the relationship between raised asparate aminotransferase (AST) and epirubicin clearance was: dose=AUC (97.5-34.2xlog AST). Adaptive dosing was evaluated prospectively in a third group of 41 women with serum AST > or =2xnormal+/-raised bilirubin. The median AUCs were 2444 and 1608 ng/ml.h, close to the high and low target AUCs, respectively. Variability in AUC was lower with adaptive dosing than in a fourth group given an unadjusted dose of epirubicin (coefficient of variation=25.8, 30.0 and 46.5%, respectively; P=0.06). Epirubicin dosing based on AST is safe and may reduce pharmacokinetic variability. PMID- 12628837 TI - Quality of life during radiotherapy for rectal cancer. AB - The aim of this study was to assess symptoms and health-related quality of life (HRQL) during (neo)adjuvant radiotherapy for rectal cancer. Patients receiving pelvic radiotherapy 50 Gy for rectal cancer, were studied prospectively (n=42). The European Organization for Research and Treatment of Cancer (EORTC) questionnaires quality of life-core 30 QLQ-C30 and QLQ-CR38 and a 5-day symptom diary were completed at the start and end of radiotherapy and 4-6 weeks later. At the end of radiotherapy, mean scores of diarrhoea, fatigue and appetite loss had significantly increased (P<0.01) compared with pretreatment scores, but this was not observed for scores for nausea or pain. At the end of radiotherapy, diarrhoea, fatigue, appetite loss, physical function, social function and global quality of life (QL) were significantly worse than the population-based norms. 64% of the patients reported an increase in fatigue and 52% an increase in diarrhoea during radiotherapy. HRQL scores had returned to pre-treatment levels 4 6 weeks after radiotherapy. Thus, diarrhoea, fatigue and appetite loss increased transiently during pelvic radiotherapy. PMID- 12628838 TI - Identification and characterisation of a group of cervical carcinoma patients with profound downregulation of intratumoral Type 1 (IFNgamma) and Type 2 (IL-4) cytokine mRNA expression. AB - Type 1 cytokines, such as interferon gamma (IFNgamma) and interleukin-2 (IL-2), increase T cell-mediated immune responses and are considered to be beneficial for antitumour immunity. Type 2 cytokines, such as IL-4, IL-5, and IL-10, inhibit Type 1 responses and promote humoral responses. We have previously reported an association between low intratumoral IFNgamma mRNA levels and poor clinical outcome in patients with invasive cervical carcinoma. In this study, by using quantitative polymerase chain reaction (PCR), we identified a group of cervical carcinoma patients with undetectable intratumoral T cell-derived cytokine mRNAs, as IFNgamma, IL-4 and IL-17 expression could not be detected in 5, 25 and 8 of the 52 biopsies analysed, respectively. Global downregulation of Type 1 and Type 2 cytokines was observed in a subgroup of patients who more frequently presented advanced stage tumours. Biopsies of patients with no IFNgamma gene expression did not appear to be less infiltrated by T cells than control biopsies with measurable IFNgamma gene expression. These results clearly demonstrate that, in some clinical situations, the decrease in intratumoral Type 1 cytokines is not associated with a Type 2 polarisation, but rather reflects global deactivation of T cells at the tumour site. These data provide support for immunotherapy protocols designed to reverse the anergic state of T cells in cancer. PMID- 12628839 TI - The role of pelvic lymph node dissection as a predictive and prognostic factor in bladder cancer. AB - The aim of this study was to evaluate the value of pelvic lymph node dissection (PLND) performed as a separate procedure in a consecutive Danish bladder cancer cohort and also to analyse if the number of lymph nodes excised had an impact on outcome. From 1992 to 1998, 339 cystectomy candidates were retrospectively reviewed. Based on a preoperative PLND, 248 patients (10% N+) underwent radical cystectomy and 91 (87% N+) underwent radio- or chemotherapy. The median follow-up was 6.3 years. PLND was able to separate N+ from N0 patients with a false negative rate of 3% compared with the following cystectomy. Lymph node-positive patients treated with cystectomy (n=24) all died from their bladder cancer. Therefore, accurate pathological N classification before the treatment decision seems worthwhile. The median number of lymph nodes excised was six and the number of lymph nodes had an independent prognostic impact on survival. This underlines the need for guidelines for surgical lymphadenectomy and the pathological assessment of lymph nodes in bladder cancer. PMID- 12628840 TI - Continuing chemotherapy or not after the induction treatment in advanced breast cancer patients. clinical outcomes and oncologists' preferences. AB - The optimal duration of cytostatic treatment for metastatic breast cancer is still a matter of debate. Possible gain in the duration of remission has to be weighed against the side-effects of treatment. Our aim was to define the optimal duration of cyclophosphamide, methotrexate, 5-fluorouracil (CMF) treatment by studying the time to treatment failure, overall survival and using a Q-TWiST analysis. The treating physician's opinion was asked. The European Organization for Research and Treatment of Cancer (EORTC) Breast Cancer Group conducted a randomised trial in 204 non-progressing metastatic breast cancer patients after induction chemotherapy (CMF) to stop or continue treatment. Progression-free (PFS) and overall survival (OS) were studied. To gain more insight into the burden of treatment-related side-effects, Q-TWiST was analysed. In addition, we asked for oncologists' preferences as patients are likely to be influenced by their physicians' opinion. Continuation of CMF had a significantly longer time to treatment failure (TTF) 5.2 versus 3.5 months (P=0.011). There was no overall survival (OS) difference 14.0 versus 14.4 months (P=0.77). Mean quality-adjusted survival time was equal to 8.4 months for no further treatment and decreased to 7.9 months for continuation of CMF (95% Confidence Interval (CI) of difference equals 0.5+/-2.5 months). Almost half of the oncologists said they would favour continuous treatment for a 3-month gain in time to progression-a difference which was not found in this study. Based on these data, an interruption of chemotherapy (CMF), if this is the wish of the patient, is justified. PMID- 12628841 TI - Biological markers that predict clinical recurrence in ductal carcinoma in situ of the breast. AB - The optimal management of ductal carcinoma in situ (DCIS) is controversial, due in part to our poor understanding of its natural history. We undertook to identify subgroups of DCIS based on the expression of biomarkers, which were related to the likelihood of clinical recurrence. Biomarker expression of a total of 95 DCIS lesions in a nested case-control study within a population-based cohort with up to 135 months follow-up data (median 101 months) was analysed using immunohistochemistry. ERBB2-positivity and bcl-2-, oestrogen receptor (ER)- and progesterone receptor (PR)-negativity were individually associated with the risk of clinical recurrence. The predictive value of these biomarkers was independent of cytonuclear grade. ERBB2, bcl-2, ER and PR expression were conserved in the recurrent lesions, including subsequent invasive cancers. p21 positive DCIS was also associated with clinical recurrence, independently of the associations with ERBB2/bcl-2/ER/PR expression. These data identify clinically and biologically relevant subcategories of DCIS lesions, an essential basis for improving management. PMID- 12628842 TI - Topoisomerase II-alpha (topoII) and HER2 amplification in breast cancers and response to preoperative doxorubicin chemotherapy. AB - A significant proportion of breast cancers with HER2 amplification have simultaneous amplification of topoisomerase II-alpha (topoII). Amplification of HER2 and topoII was assayed using a novel chromogenic in situ hybridisation (CISH) method. HER2 and topoII amplification status and the response to preoperative doxorubicin chemotherapy were analysed in 67 locally advanced breast cancer patients. Response to chemotherapy was increased in the cases with coamplification of HER2 and topoII (18/19), whereas the response rate was significantly decreased in the cases without HER2 and topoII amplification (17/36). The 12 cases with HER2 amplification alone showed an intermediate response rate (9/12). The findings of the current study indicate that topoII amplification may play a role in determining chemosensitivity of breast cancers to doxorubicin chemotherapy. PMID- 12628843 TI - Mannose-6-phosphate/insulin-like growth factor-II receptor expression levels during the progression from normal human mammary tissue to invasive breast carcinomas. AB - The putative role of mannose-6-phosphate/insulin-like growth factor-II receptor (M6P/IGFII-R) as a tumour suppressor and its value as a prognostic marker of breast cancer was studied in 42 benign breast diseases (BBD), 61 in situ carcinomas (CIS) and 133 invasive carcinomas. The receptor was quantified by immunohistochemistry with a computerised image analyser, using specific polyclonal IGY antibodies. The M6P/IGFII-R level varied markedly according to the different patient samples, but median values and distributions were similar in lesions and normal adjacent glands. However, the receptor level was significantly increased in high-grade ductal carcinomas in situ (DCIS) and decreased in invasive carcinomas relative to adjacent normal tissue. The M6P/IGFII-R protein concentration in invasive breast carcinomas was mostly independent of prognostic parameters: tumour size, histological grade, lymph node (N) invasiveness and oestrogen receptor alpha (ERalpha) status. The only positive correlation was with cathepsin D, the progesterone receptor (PgR) and with patients aged >60 years. These results do not support the hypothesis of a frequent and early inactivation of the M6P/IGFII-R gene in breast cancer. Clinical follow-up of patients might reveal a prognostic value for one of the cathepsin receptors. PMID- 12628844 TI - Management of infection in children with malignancy. PMID- 12628845 TI - Infections in children with cancer-old approaches and new. PMID- 12628846 TI - Differences in the epidemic rise and decrease of prostate cancer among geographical areas in Southern Europe. an analysis of differential trends in incidence and mortality in France, Italy and Spain. AB - This is a population-based study aimed at evaluating incidence and mortality trends for prostate cancer in France, Italy and Spain, during the prostate specific antigen (PSA) era, considering elderly people aged 70 years and over and younger adults aged between 40 and 69 years. Trends were estimated by a log linear Poisson regression model and expressed as an Estimated Annual Percent Change (EAPC). Incidence increased sharply in almost all areas. Spain showed the lowest increases. Incidence started to rise around 1985 in France and after 1990 in Italy and Spain. Mortality increased until the late 1980s in all countries, then declined in France and Italy (-2.5% in 40-69 year age group), but not in Spain. Younger people showed a much higher rise in incidence than the elderly, while mortality decreased mainly in the younger adults. The decrease in mortality was more marked in those areas and the younger age group where the rise in incidence was higher and started earlier, i.e. in France and in younger people, suggesting that the PSA test may have had a positive effect on mortality, although other clinical advances also have to be taken into account. PMID- 12628847 TI - Neovascularisation, expression of fibroblast growth factor-2, and mast cells with tryptase activity increase simultaneously with pathological progression in human malignant melanoma. AB - Tissues from 92 proliferative lesions of the melanocytic lineage defining distinct steps in tumour progression were investigated immunohistochemically for changes in angiogenesis, expression of fibroblast growth factor-2 (FGF-2) and density of total mast cells (MCs) and MCs expressing tryptase, an angiogenic inducing molecule. Although the microvessel number was low in common nevi, it increased significantly in nevi with architectural disorder with varying degrees of melanocytic atypia (termed 'nevi with ADMA'), and these changes persisted during tumour development. Progression of primary melanomas was accompanied by a high microvessel number, and the progression to metastases by another significant increase in the microvessel counts. Expression of FGF-2, evaluated as percentages of positive lesions and positive cells per lesion was upregulated in the course of progression. Changes in expression were associated with nevi with ADMA, tumour changeover, penetration of the tumour into the dermis and metastases. A high correlation was demonstrated in all groups of tissues between the microvessel counts, percentages of FGF-2-positive tumour cells, and both total metachromatic and tryptase-reactive MCs. These results suggest that angiogenesis in human melanoma increases with tumour progression and that FGF-2 secreted by tumour cells and tryptase secreted by host MCs cooperate in its induction. PMID- 12628848 TI - Predicting the maximum-tolerated dose of PNU-159548 (4-demethoxy-3'-deamino-3' aziridinyl-4'-methylsulphonyl-daunorubicin) in humans using CFU-GM clonogenic assays and prospective validation. AB - A haematotoxicity model was proposed by Parchment in 1998 to predict the maximum tolerated dose (MTD) in humans of myelosuppressive antitumour agents by combining data from in vitro clonogenic assays on haematopoietic progenitors and in vivo systemic exposure data in animals. A prospective validation of this model in humans was performed with PNU-159548, a novel agent showing selective dose limiting myelosuppression in animals. PNU-159548 and its main metabolite, PNU 169884, were tested in vitro on murine, canine and human colony forming units granulocyte macrophages (CFU-GM) and in vivo on mice and dogs. The IC(90x) ratios (IC(x)=concentration inhibiting x% of colony growth) for CFU-GM and drug plasma protein binding were used to adjust the target plasma concentrations versus time curve (AUC) and predict the human MTD. The predicted MTD was compared with values achieved in phase I studies. Canine CFU-GM were 6-fold more sensitive (P<0.01) and murine CFU-GM 1.7-fold less sensitive (P<0.05) to PNU-159548 treatment than the human progenitors. PNU-169884 behaved similarly to PNU-159548. The predicted MTDs in humans calculated from data in mice and dogs were 15 and 38 mg/m(2), respectively. Overall, 61 patients were treated in two phase I studies, at doses ranging from 1.0 to 16 mg/m(2). Thrombocytopenia was dose-limiting with a MTD of 14 and 16 mg/m(2) in heavily and minimally pretreated/non-pretreated patients, respectively. Adjusting animal MTD data by means of the CFU-GM ratio between species can predict the human MTD with a good quantitative accuracy. Inhibition of common haemopoietic progenitors by PNU-159548 induced neutropenia/thrombocytopenia in animals and thrombocytopenia in patients, probably due to the higher sensitivity to the compound observed in human colony forming units-megakaryocyte (CFU-MK). PMID- 12628849 TI - Detection of the human papillomavirus and analysis of the TP53 polymorphism of exon 4 at codon 72 in penile squamous cell carcinomas. AB - Human papillomaviruses (HPV) are thought to be involved in penile squamous cell carcinomas (SCC). A common polymorphism at codon 72 of exon 4 encoding either arginine (Arg) or proline (Pro) has been shown to affect HPV-mediated degradation of p53 in vitro, and may represent a risk factor for HPV-induced carcinogenesis. The presence of HPV DNA as well as the TP53 polymorphism at codon 72 of exon 4 were investigated in a series of 45 penile SCC. HPV detection and typing were carried out by polymerase chain reaction (PCR) with generic primers (MY09-MY11 and FAP59-FAP64), and type-specific DNA probes. TP53 polymorphism was further investigated using Denaturing Gradient Gel Electrophoresis (DGGE). HPV DNA was detected in 67% of penile SCC and 32% of benign lesions (BL) (P<0.05). Among the TP53 amplified samples, the rate of Arg homozygosity in penile SCC was 61% compared with 68% in BL (non-significant (NS)). Our results demonstrate a strong association between penile SCC and the presence of HPV DNA. The TP53 Arg/Arg genotype does not appear to represent a risk factor for the development of genital SCC in men, and no correlation was found between the TP53 polymorphism at codon 72 and the presence of HPV DNA. PMID- 12628850 TI - Expression of deoxycytidine kinase in leukaemic cells compared with solid tumour cell lines, liver metastases and normal liver. AB - Deoxycytidine kinase (dCK) is required for the phosphorylation of several deoxyribonucleoside analogues that are widely employed as chemotherapeutic agents. Examples include cytosine arabinoside (Ara-C) and 2-chlorodeoxyadenosine (CdA) in the treatment of acute myeloid leukaemia (AML) and gemcitabine to treat solid tumours. In this study, expression of dCK mRNA was measured by a competitive template reverse transcriptase polymerase chain reaction (CT RT-PCR) in seven cell lines of different histological origin, 16 childhood and adult AML samples, 10 human liver samples and 11 human liver metastases of colorectal cancer origin. The enzyme activity and protein expression levels of dCK in the cell lines were closely related to the mRNA expression levels (r=0.75, P=0.026 and r=0.86, P=0.007). In AML samples, dCK mRNA expression ranged from 1.16 to 35.25 (x10(-3)xdCK/beta-actin). In the cell line panel, the range was 2.97-56.9 (x10(-3)xdCK/beta-actin) of dCK mRNA expression. The enzyme activity in liver metastases was correlated to dCK mRNA expression (r=0.497, P=0.05). In the liver samples, these were not correlated. dCK mRNA expression showed only a 36-fold range in liver while a 150-fold range was observed in the liver metastases. In addition, dCK activity and mean mRNA levels were 2.5-fold higher in the metastases than in the liver samples. Since dCK is associated with the sensitivity to deoxynucleoside analogues and because of the good correlation between the different dCK measurements in malignant cells and tumours, the CT-RT PCR assay will be useful in the selection of patients that can be treated with deoxycytidine analogues. PMID- 12628852 TI - Chronic cough: practical considerations. PMID- 12628853 TI - The cough reflex and the "healthy smoker". PMID- 12628851 TI - Establishment and characterisation of human papillomavirus type 16 DNA immortalised human tonsillar epithelial cell lines. AB - Oncogenic human papillomavirus (HPV) plays a possible aetiological role in a subset of head and neck cancers, particularly in tonsillar carcinomas. For establishing a model to study mechanisms involved in HPV-associated tonsillar carcinogenesis, normal human tonsillar epithelial (HTE) cells were transfected with full-length HPV-16 DNA. The transfections produced four immortalised cell lines, designated HTE-114/K1, HTE-114/K2, HTE-114/K3 and HTE-114/B. All transfected HTE cell lines were cytogenetically abnormal. They exhibited altered morphology and impaired expression of cytokeratins in organotypic cultures. They failed to form colonies in soft agarose and formed no tumours in nude mice within 6 months. Each of them contained integrated viral DNA in a distinctive pattern as shown by Southern blot hybridisation. Early viral transcripts containing the E7 gene were detected by northern blot hybridisation. In conclusion, primary HTE cells can be immortalised following transfection with full-length HPV-16 DNA; the immortalised cell lines had partially retained epithelial characteristics in their morphology and function. They seem to represent early stages of premalignant epithelial cells and thus provide a useful model for studying further the multistep molecular events of HPV-16-associated tonsillar carcinogenesis. PMID- 12628854 TI - Continuous positive airway pressure: by nose or mouth? PMID- 12628855 TI - What is "heart rate variability" and is it blunted by tumor necrosis factor? PMID- 12628856 TI - What is the best way to measure lung function? PMID- 12628857 TI - Urine color test to monitor isoniazid compliance: "pissin' in the wind"? PMID- 12628858 TI - Antibiotic therapy of ventilator-associated pneumonia: in search of the magic bullet. PMID- 12628859 TI - Heliox redux. PMID- 12628860 TI - Heliox for status asthmaticus? PMID- 12628861 TI - Clinician or statistician? PMID- 12628862 TI - Chronic cough and gastroesophageal reflux disease: experience with specific therapy for diagnosis and treatment. AB - STUDY OBJECTIVE: To evaluate experience using a therapeutic trial of proton-pump inhibitor therapy with or without a prokinetic agent in diagnosis and treatment of gastroesophageal reflux disease (GERD)-related cough. DESIGN: A review of experience with 214 patients with cough of > or = 3 weeks referred over 3.5 years. An anatomic diagnostic protocol was used to identify and treat those with GERD-related cough. SETTING: A pulmonary specialty practice affiliated with the University of Rochester School of Medicine and Dentistry. PATIENTS: One hundred eighty-three patients were identified with chronic cough and were included in the study. Thirty-one patients were disqualified because of abnormal chest radiographic findings, inadequate follow-up, or cough being not the primary complaint. Fifty-six patients were identified as having GERD-related cough. INTERVENTIONS: A once-daily dose of a proton-pump inhibitor was prescribed. A prokinetic agent was added if esophageal dysfunction was suspected or response was inadequate. Those who did not respond underwent 24-h esophageal pH monitoring. RESULTS: GERD was the single cause of cough in 24 patients (43%). Twenty-nine patients (52%) had GERD plus another cause, and 3 patients (5%) had GERD with more than two causes. Twenty-four patients (43%) had cough only, while 32 patients (57%) had other symptoms of GERD. Proton-pump therapy was successful in 42 patients (79%). Twenty-four patients responded to proton-pump inhibitor therapy, and 18 patients responded when metoclopramide or cisapride was added. The remaining two patients responded to a histamine type-2 blocker or cisapride alone. The cough was eliminated or markedly improved in 38 patients (86%) after 4 weeks and by 8 weeks in the remaining 6 patients. Six of the nonresponders had aspiration diagnosed by bronchoscopy. Four patients had fundoplication recommended, and two patients responded to alternative interventions. CONCLUSIONS: Four to 6 weeks of a proton-pump inhibitor alone or in combination with a prokinetic agent successfully diagnoses and treats four of five patients with GERD-related cough. Twenty-four-hour esophageal pH monitoring will confirm the diagnosis in the others. These patients may be candidates for fundoplication. Nonresponders often aspirate as an additional aggravating factor. PMID- 12628863 TI - Cough reflex sensitivity in cigarette smokers. AB - STUDY OBJECTIVES: To evaluate cough reflex sensitivity in a population of young, healthy, male cigarette smokers. DESIGN: Cross-sectional comparison. SETTING: Academic medical center. PARTICIPANTS: Twenty healthy, male current-smokers (mean [+/- SEM] age, 32.2 +/- 1.2 years). MEASUREMENTS: Subjects underwent baseline spirometry followed by capsaicin cough challenge testing, which involved the inhalation of capsaicin in ascending, doubling concentrations until the concentrations inducing two or more coughs (C(2)) and those inducing five or more coughs (C(5)) were reached. The data were compared to those from a group of 50 healthy, male nonsmokers who had undergone identical cough challenge testing. RESULTS: The two groups did not differ in terms of age or baseline pulmonary function. Cough sensitivity was significantly diminished in the current-smokers compared to control subjects. The mean (+/- SEM) log C(2) values in smokers and nonsmokers were 1.26 +/- 0.13 and 0.81 +/- 0.08, respectively (p = 0.004). The mean log C(5) values in smokers and nonsmokers were 2.03 +/- 0.10 and 1.20 +/- 0.08, respectively (p < 0.000001). CONCLUSIONS: Cough reflex sensitivity is significantly diminished in young, healthy, male current-smokers compared to a similar population of nonsmokers. The mechanism of cough suppression in smokers remains speculative but may involve long-term tobacco smoke-induced desensitization of the cough receptors within the airway epithelium. PMID- 12628864 TI - A physiologic comparison of nasal and oral positive airway pressure. AB - STUDY OBJECTIVES: The effectiveness of nasal continuous positive airway pressure (CPAP) in treating obstructive sleep apnea (OSA) is based on raising the intramural pressure above a critical collapsing pressure of the oropharyngeal airway. It is currently unclear whether CPAP delivered orally is also capable of raising pressure in the oropharynx above the critical collapse pressure. DESIGN: We tested a novel oral CPAP device to determine whether the pressure-flow relationships are similar to nasal CPAP and whether the device alters these relationships. Patients were selected based on having moderately severe apnea and were randomized to nasal CPAP, nasal CPAP with oral device, or oral CPAP. SETTING: Johns Hopkins University, The Johns Hopkins Asthma and Allergy Center, Baltimore, MD. PATIENTS: Five men and two women with OSA were studied. INTERVENTIONS: Individual pressure-flow curves were constructed during the application of nasal or oral CPAP. RESULTS: We found the following: (1) a similar effective pressure eliminated inspiratory flow limitation for the nasal or oral CPAP; (2) as pressure in the nose or mouth was lowered below the effective pressure, a linear pressure-flow curve was obtained and a critical closing pressure was described; (3) similar mean (+/- SD) critical pressures of -0.3 +/- 5.3, 1.7 +/- 4.0, and 0.5 +/- 2.8 cm H(2)O, respectively, occurred for nasal CPAP, nasal CPAP with the oral device in place, and oral CPAP conditions (p > 0.1); and (4) the comparable mean values for upstream resistance were 27.8 +/- 19, 19.1 +/- 8.3, and 26.5 +/- 26.7 cm H(2)O/L/s, respectively, for the above three conditions (p > 0.1). CONCLUSIONS: We concluded that comparable upper airway pressure-flow relationships were obtained during oral and nasal breathing. Moreover, effective treatment pressure is obtained when constant pressure is applied through either the nasal or oral route. PMID- 12628865 TI - Evaluation of a portable device based on peripheral arterial tone for unattended home sleep studies. AB - BACKGROUND: Diagnosis of obstructive sleep apnea syndrome (OSAS) by ambulatory systems is a growing practice in view of the large number of patients awaiting correct diagnosis. The Watch PAT100 (WP100) [Itamar Medical; Caesarea, Israel] is a portable device based on the peripheral arterial tone (PAT) signal, and is designed for unattended home sleep studies. OBJECTIVES: To evaluate the efficacy, reliability, and reproducibility of the WP100 device for the diagnosis of OSAS as compared to in-laboratory, standard polysomnographic-based manual scoring. DESIGN AND METHODS: One hundred two subjects (78 men; 69 patients with OSAS and 33 normal volunteers; mean +/- SD age, 41.4 +/- 15.2 years; body mass index, 26.8 +/ 5.5) underwent in-laboratory full polysomnography simultaneously with WP100 recording. Fourteen subjects also underwent two additional unattended home sleep studies with the WP100 alone. The polysomnography recordings were blindly scored for apnea/hypopnea according to the American Academy of Sleep Medicine criteria (1999), and the polysomnography respiratory disturbance index (RDI) [PSG-RDI] was calculated. The WP100 data were analyzed automatically for the PAT RDI (PRDI) by a proprietary algorithm that was previously developed on an independent group of subjects. RESULTS: Across a wide range of RDI levels, the PRDI was highly correlated with the PSG-RDI (r = 0.88, p < 0.0001), with an area under the receiver operating characteristic curve of 0.82 and 0.87 for thresholds of 10 events per hour and 20 events per hour, respectively. The PRDI scores were also highly reproducible, showing high correlation between home and in-laboratory sleep studies (r = 0.89, p < 0.001). CONCLUSION: The WP100 may offer an accurate, robust, and reliable ambulatory method for the detection of OSAS, with minimal patient discomfort. PMID- 12628866 TI - Evaluation of unattended automated titration to determine therapeutic continuous positive airway pressure in patients with obstructive sleep apnea. AB - BACKGROUND: Determination of the therapeutic pressure during continuous positive airway pressure (CPAP) therapy is usually performed by a technician during polysomnography. In recent years, several devices for automated adjustment of the therapeutic pressure by the means of computerized algorithms were developed. The aims of the present study were to compare two different devices for automated titration and to verify if unattended automated titration is a feasible strategy to determine the therapeutic CPAP. METHODS: We enrolled 16 consecutive patients with obstructive sleep apnea syndrome (OSAS) defined by an apnea-hypopnea index > 20/h. Automated titration was performed in the hospital using two CPAP devices (Autoset; Resmed; North Ryde, Australia; and Somnosmart; Weinmann; Hamburg, Germany) in random order for 2 consecutive nights, based on different signals for the detection of respiratory events. During titration, there was no direct supervision by a technician, and polysomnography was not recorded. We defined the therapeutic pressure as the 95th percentile of the airway pressure over time (P95). RESULTS: We observed significant differences of the P95 between the two devices, with an average of 7.0 +/- 2.5 cm H(2)O for the Somnosmart and 9.9 +/- 2.6 cm H(2)O for the Autoset (p = 0.005) [mean +/- SD]. There was a considerable lack of agreement between the two devices, with a bias of 3.0 cm H(2)O and limits of agreement ranging from + 9.3 to - 3.2 cm H(2)O. We found no significant correlation between the paired differences of P95 and either indexes of severity of OSAS or lung function variables. CONCLUSION: Automated titration based on the analyses of flow (Autoset) or forced oscillations (Somnosmart) predicted significant different therapeutic pressures for fixed CPAP therapy. Thus, unattended automated titration performed during 1 night of hospital stay with commercially available devices cannot be used to determine accurately the therapeutic CPAP in patients with OSAS. PMID- 12628867 TI - Unexplained pulmonary hypertension is associated with systolic arterial hypertension in patients undergoing routine Doppler echocardiography. AB - STUDY OBJECTIVE: To determine the validity of the association between systemic hypertension (HTN) and unexplained pulmonary hypertension (PHTN) as identified with Doppler echocardiography. METHODS: All patients with a reported systolic pulmonary artery pressure (SPAP) on routine Doppler echocardiography from our 1997 echocardiographic database were identified. Exclusions included all diseases known to be associated with PHTN. Of 1,174 patients, 503 had PHTN (defined as a SPAP of >/= 40 mm Hg), of whom 42 (8.4%) had unexplained PHTN. These PHTN patients were matched for age (mean [+/- SD] age, 70 +/- 11 years) with 84 randomly selected patients from the same database who had normal SPAP values and no diseases associated with PHTN. RESULTS: The mean SPAP of those patients with unexplained PHTN was 48 +/- 9 mm Hg vs 31 +/- 5 mm Hg for those without unexplained PHTN. HTN was more prevalent in those with PHTN (98% vs 72%, respectively; p = 0.0008). Patients with unexplained PHTN had significantly higher mean systolic BP, as routinely measured at the end of the echo (154 +/- 26 vs 138 +/- 21 mm Hg, respectively; p = 0.0006), but they did not differ in diastolic BP (80 +/- 14 vs 78 +/- 11 mm Hg, respectively; p = 0.39). PHTN patients and control subjects did not differ with respect to gender (women, 74% vs 70%, respectively), race (white, 64% vs 65%, respectively), body mass index (30 +/- 8 vs 28 +/- 8 kg/m(2), respectively), or left ventricular ejection fraction (64 +/- 6% vs 63 +/- 7%, respectively). When only those with known HTN were considered, PHTN patients still had higher systolic arterial BP (155 +/- 25 vs 143 +/- 21 mm Hg, respectively; p = 0.013) and tended to be on more BP medications (1.6 +/- 1.1 vs 1.2 +/- 0.9, respectively; p = 0.09). CONCLUSIONS: Unexplained PHTN occurs mostly in the elderly, is associated with systolic HTN, and those hypertensive patients with concomitant PHTN have higher systolic arterial pressures. PMID- 12628868 TI - Circulating levels of tumor necrosis factor correlate with indexes of depressed heart rate variability: a study in patients with mild-to-moderate heart failure. AB - OBJECTIVES: Patients with heart failure have increased circulating levels of tumor necrosis factor (TNF) and TNF receptors. It is not known whether TNF, which is known to blunt beta-adrenergic responsiveness in experimental models, contributes to the loss of heart rate variability in patients with heart failure. Therefore, we examined heart rate variability in relation to circulating levels of TNF, TNF receptors, and norepinephrine in patients with heart failure and in control subjects. METHODS: Heart rate variability was obtained from 24-h ambulatory ECG recordings in age-matched control subjects (n = 10) and patients with mild (n = 15) to moderate (n = 14) heart failure. Plasma levels of TNF and soluble type 1 and 2 TNF receptors were measured by enzyme-linked immunoassay; plasma norepinephrine levels were measured by high-performance liquid chromatography. RESULTS: There was a significant inverse linear correlation between increased circulating levels of TNF, TNF receptors, and norepinephrine for time-domain and frequency-domain indexes of heart rate variability among patients with heart failure and control subjects. Multiple stepwise linear regression analysis showed that TNF was a stronger independent predictor of frequency-domain indexes of heart rate variability than norepinephrine. CONCLUSIONS: TNF is an independent predictor of depressed heart rate variability in patients with heart failure. Insofar as TNF blunts beta-adrenergic signaling, this study suggests the possibility that overexpression of TNF and subsequent loss of beta-adrenergic responsiveness contributes to the decrease in heart rate variability observed in heart failure. PMID- 12628869 TI - Spirometry testing standards in spinal cord injury. AB - STUDY OBJECTIVES: Because muscle paralysis makes it uncertain whether subjects with spinal cord injury (SCI) can perform spirometry in accordance with American Thoracic Society (ATS) standards, determinants of test failure were examined. DESIGN: Cross-sectional study. SETTING: Veterans Affairs (VA) medical center. PARTICIPANTS: Veterans with SCI at VA Boston Healthcare System and nonveterans recruited by mail and advertisement. MEASUREMENTS AND RESULTS: Two hundred thirty of 278 subjects (83%) were able to produce three expiratory efforts lasting > or = 6 s and without excessive back-extrapolated volume (EBEV). In 217 of 230 subjects (94%), FVC and FEV(1) were each reproducible in accordance with 1994 ATS standards. In the remaining 48 subjects, efforts with smooth and continuous volume-time tracings and acceptable flow-volume loops were identified. These subjects had a lower percentage of predicted FVC, FEV(1), and maximum expiratory and inspiratory pressures compared to the others, and a greater proportion had neurologically complete cervical injury (42% compared to 16%). In 19 subjects (40%), some expiratory efforts were not sustained maximally for > or = 6 s but had at least a 0.5-s plateau at residual volume (short efforts). In eight subjects (17%), some efforts were not short but had EBEV. In the remaining 21 subjects (44%), some efforts were short, some had EBEV, and some had both. If these efforts were not rejected, 262 of 278 subjects (94%) would have produced three acceptable efforts, and in 257 subjects (92%), the efforts were reproducible. CONCLUSIONS: Subjects with SCI with the most impaired respiratory muscles and abnormal pulmonary function are able to perform spirometry reproducibly despite not meeting usual ATS acceptability standards. Exclusion of these subjects would lead to bias in studies of respiratory function in SCI. The modification of spirometry testing standards to include efforts with EBEV and with a 0.5-s plateau if < 6 s would reduce the potential for bias. PMID- 12628870 TI - Comparison of maximal midexpiratory flow rate and forced expiratory flow at 50% of vital capacity in children. AB - BACKGROUND: The mid-portion of the maximal expiratory flow-volume (MEFV) curve is often described by values of the mean forced expired flow as lung volume decreases from 75% to 25% of vital capacity (ie, forced expiratory flow, midexpiratory phase [FEF(25-75)]). It is common practice to report also forced expired flow at 50% of vital capacity (FEF(50)). STUDY OBJECTIVE: To investigate whether FEF(50) and FEF(25-75) are highly correlated or whether the difference between them reflects a degree of airways obstruction. Also, we wanted to investigate the correlation between the two in cases of irregularly shaped MEFV curves (ie, "saw-toothing"). DESIGN: Analysis of the correlation between FEF(50) and FEF(25-75) in a single determination. We assessed the relationship between the FEF(50)/FEF(25-75) ratio and the degree of airways obstruction, as reflected by other traditional parameters such as FEV(1), FEV(1)/FVC ratio, and specific airway conductance (SGaw). PATIENTS: There were 1,350 forced expiratory maneuvers performed by children with a broad range of pulmonary abnormalities. RESULTS: FEF(50) correlated with FEF(25-75) as follows: FEF(50) (L/s) = 0.041 + 1.136*FEF(25-75)(L/s); r(2) = 0.956; standard error of the estimate = 0.013; p < 0.0001. The FEF(50)/FEF(25-75) ratio remained stable and did not correlate with FEV(1) (r = 0.12), FEV(1)/FVC ratio (r = 0.11), or SGaw (r = 0.02; difference not significant). The correlation between FEF(25-75) and FEF(50) was similar for both the smooth curve (r = 0.97) and the irregular curve (r = 0.96). CONCLUSIONS: Although not identical, FEF(25-75) and FEF(50) are highly correlated, and the ratio of the two is fairly constant. Therefore, the practice of reporting both of them is unnecessary. We suggest that it is reasonable to prefer FEF(50). PMID- 12628871 TI - Use of the urine color test to monitor compliance with isoniazid treatment of latent tuberculosis infection. AB - STUDY OBJECTIVE: To apply the Arkansas color method in order to evaluate drug compliance and factors that can predict treatment adherence in patients being treated for latent tuberculosis infection (LTBI) with a single daily dose of isoniazid (INH). DESIGN: Prospective study of adherence of 105 patients aged 1 to 75 years who were treated with a single daily dose of INH for LTBI. INTERVENTIONS: Patients or their parents were interviewed regarding parameters that may affect compliance. Urine samples were collected and tested for INH metabolites with the Arkansas color method. RESULTS: Nonadherence to treatment was found in 28.5% of patients. There was no statistically significant correlation among the following parameters: gender; age; diagnosis; mode of administration (self or parents); duration of treatment; dose of INH per weight; or interval since last intake of dose. Twenty-six patients were randomly checked for treatment adherence on two separate visits, and nonadherent patients were informed immediately and their condition was fully explained to them. Five of six patients who were nonadherent in the first visit and were examined twice became adherent in the second visit. Three of 20 patients who were adherent in the first visit became nonadherent. CONCLUSION: Almost one third of the patients who received LTBI treatment with INH were nonadherent to treatment. No factor was found to predict adherence. The Arkansas method can be used by the family physician and is a simple, immediate method to follow-up patients with LTBI who are treated with INH. PMID- 12628872 TI - Diagnostic significance of interferon-gamma in tuberculous pleural effusions. AB - STUDY OBJECTIVES: Tuberculosis (TB), the single most frequent infectious cause of death worldwide, also is a major cause of pleural effusion, which in TB usually has lymphocytic and exudative characteristics. Differential diagnosis between TB and nontuberculous pleural effusion can be sometimes difficult, representing a critically important clinical problem. METHODS: We studied 46 patients presenting with pleural effusion to the National Sanyo Hospital between April 2000 and January 2001 (34 men and 12 women; mean age, 64 years). Ten patients (22%) had tuberculous pleurisy, 19 patients (41%) had malignant pleuritis, and 17 patients (37%) had pleural effusion due to an etiology other than tuberculosis or cancer. Pleural fluid concentrations of four suggested markers were measured using commercially available kits. RESULTS: The pleural fluid levels (mean +/- SE) of adenosine deaminase (83.3 +/- 18.2 U/L vs 25.8 +/- 20.4 U/L, p < 0.0001), interferon-gamma (137 +/- 230 IU/mL vs 0.41 +/- 0.05 IU/mL, p < 0.0001), immunosuppressive acidic protein (741 +/- 213 micro g/mL vs 445 +/- 180 micro g/mL, p < 0.001) and soluble interleukin 2 receptor (7,618 +/- 3,662 U/mL vs 2,222 +/- 1,027 U/mL, p < 0.0001) were significantly higher for tuberculous pleuritis than for other causes of effusion. Receiver operating characteristic analysis demonstrated that pleural fluid content INF-gamma was the best indicator of tuberculous pleurisy among four relevant biological markers. CONCLUSIONS: INF gamma in pleural fluid is the most sensitive and specific among four biological markers for tuberculous pleuritis. Thus, our results suggest that determination of INF-gamma at the onset of pleural effusion is informative for the diagnosis of tuberculous pleuritis. Further studies including larger numbers of patients are needed to verify this result. PMID- 12628873 TI - The role of fungal spores in thunderstorm asthma. AB - STUDY OBJECTIVES: To document the existence and investigate the etiology of "thunderstorm asthma," which has been reported sporadically over the past 20 years. DESIGN: We assessed the relationship between thunderstorms, air pollutants, aeroallergens, and asthma admissions to a children's hospital emergency department over a 6-year period. RESULTS: During thunderstorm days (n = 151 days) compared to days without thunderstorms (n = 919 days), daily asthma visits increased from 8.6 to 10 (p < 0.05), and air concentrations of fungal spores doubled (from 1,512 to 2,749/m(3)), with relatively smaller changes in pollens and air pollutants. Daily time-series analyses across the 6 years of observation, irrespective of the presence or absence of thunderstorms, demonstrated that an increase in total spores, equivalent to its seasonal mean, was associated with a 2.2% (0.9% SE) increase in asthma visits. CONCLUSIONS: Our results support a relationship between thunderstorms and asthma, and suggest that the mechanism may be through increases in spores that exacerbate asthma. Replication in other climates is suggested to determine whether these findings can be generalized to other aeroallergen mixes. PMID- 12628874 TI - Prospective evaluation of the validity of exhaled nitric oxide for the diagnosis of asthma. AB - STUDY OBJECTIVE: Exhaled nitric oxide (NO) levels are significantly elevated in patients with inflammatory airways disorders such as asthma, and the measurement of exhaled NO has been proposed as a noninvasive marker of airways inflammation. The aim of this study was to assess the accuracy of exhaled NO levels for the diagnosis of asthma. METHODS: Two hundred forty consecutive, nonsmoking, steroid naive patients, who were referred to our outpatient clinic with symptoms suggestive of obstructive airways disease, were investigated. Asthma was diagnosed in 160 patients on the basis of the presence of significant airways reversibility (DeltaFEV(1) > 12% predicted) and/or airways hyperresponsiveness (provocative concentration of histamine causing a 20% fall in FEV(1) < or = 8 mg/mL). Prior to lung function measurements, exhaled NO was measured during a single-breath exhalation, according to European Respiratory Society and American Thoracic Society guidelines. RESULTS: The measurement of exhaled NO in our study population showed, at a cutoff level of 16 parts per billion, a specificity for the diagnosis of asthma of 90% and a positive predictive value of > 90%. CONCLUSIONS: These findings suggest that the simple and absolutely noninvasive measurement of exhaled NO can be used as an additional diagnostic tool for the screening of patients with a suspected diagnosis of asthma. PMID- 12628875 TI - Concomitant chronic sinusitis treatment in children with mild asthma: the effect on bronchial hyperresponsiveness. AB - STUDY OBJECTIVE: Previous studies have suggested that aggressive treatment of sinusitis can decrease bronchial hyperresponsiveness (BHR). However, there is still too little evidence to draw this conclusion, and the concept remains controversial. DESIGN: A prospective, open-label study. SETTING: University children's hospital allergy and immunology center and radiologic department. PATIENTS: Sixty-one children with mild asthma and allergic rhinitis participated in the study. Forty-one of these 61 children had sinusitis, and the remainder had no sinusitis. Ten matched, nonatopic, healthy children were used as a control group. INTERVENTION: Children with chronic sinusitis were placed into two groups. One group was treated with amoxicillin-clavulanate for 6 weeks and then with nasal saline solution irrigation for 6 weeks. For the other group, the treatment order was reversed. Children without chronic sinusitis received nasal saline solution irrigation for 12 weeks. MEASUREMENTS: Clinical symptoms and signs of sinusitis, FEV(1), and BHR were analyzed in the patients before and after treatment. RESULTS: The clinical symptoms and signs of sinusitis, but not FEV(1), showed a significant improvement after antibiotic treatment. After aggressive treatment for sinusitis, it was found that the provocative concentration of methacholine causing a 20% fall in FEV(1) of children with mild asthma and sinusitis was significantly higher after treatment. CONCLUSION: The results suggest that every asthmatic patient needs to carefully evaluate to determine whether the patient has concomitant sinusitis. Respiratory infections that meet criteria for sinusitis, even if they do not exacerbate asthma, should be treated. It is suggested that sinusitis should always be kept in mind as a possible inducible factor for BHR, and that aggressive treatment of chronic sinusitis is indicated when dealing with an asthmatic patient who shows an unpredictable response to appropriate treatment. Moreover, the findings of this study provide more evidence for an association between sinusitis and asthma with respect to BHR. PMID- 12628877 TI - Efficacy and safety of azithromycin vs levofloxacin in the outpatient treatment of acute bacterial exacerbations of chronic bronchitis. AB - STUDY OBJECTIVES: To compare the safety and efficacy of oral azithromycin and levofloxacin in the treatment of outpatients with acute bacterial exacerbations of chronic bronchitis (ABECB). DESIGN: Randomized, double-blinded, double-dummy, multicenter trial with 1:1 treatment allocation. SETTING: Outpatient treatment setting. PATIENTS: Two hundred thirty-five male or female outpatients between the ages of 35 and 75 years who had received a clinical diagnosis of ABECB. INTERVENTIONS: Blinded treatment with either oral azithromycin, 500 mg on day 1 and 250 mg per day for days 2 to 5, or, oral levofloxacin, 500 mg q24h for 7 days. RESULTS: Both treatments were well-tolerated, with the majority of adverse events being GI in nature. Favorable clinical outcomes in clinically evaluable patients were demonstrated in 89% of patients receiving azithromycin and in 92% of patients receiving levofloxacin by day 4 of therapy. At day 24, the posttherapy visit, favorable responses were approximately 82% and 86%, respectively, for patients in the two treatment groups. The bacterial eradication rates of respiratory pathogens were 96% for azithromycin and 85% for levofloxacin. CONCLUSIONS: Despite increasing concerns over macrolide resistance and a higher incidence of Gram-negative pathogens, a standard 5-day course of oral azithromycin was clinically and bacteriologically equivalent to a 7-day course of oral levofloxacin in the treatment of patients with ABECB. PMID- 12628876 TI - Asthma features in severe alpha1-antitrypsin deficiency: experience of the National Heart, Lung, and Blood Institute Registry. AB - STUDY OBJECTIVES: To describe asthma features in a cohort with alpha(1) antitrypsin (AAT) deficiency, and determine the impact of asthma on FEV(1) decline. BACKGROUND: Asthma may be common in those with AAT deficiency, and may lead to accelerated airflow obstruction. DESIGN: Analysis of data obtained from a 5-year, prospective National Heart, Lung, and Blood Institute registry. SETTING: A multicenter registry consisting of 37 clinical centers, a central phenotyping laboratory, and a data analysis center. PARTICIPANTS: A cohort of 1,052 subjects with AAT deficiency. MEASUREMENTS AND RESULTS: Asthma was defined as reversible airflow obstruction, recurrent attacks of wheezing, and a reported diagnosis of asthma or allergy with or without an elevated serum IgE level. FEV(1) decline was calculated by least-square means with adjustments for covariables. Asthma was present in 21% of the cohort and in 12.5% of those with a normal FEV(1). Attacks of wheezing were reported in 66%, the first attack occurring at a mean +/- SD age of 31 +/- 16 years. Allergy and asthma was reported in 29% and 38%, respectively. An elevated IgE level occurred in 17% and was significantly associated with signs and symptoms of asthma and an allergy history. Unadjusted FEV(1) decline was less in the group without asthma and a normal IgE level (- 48.5 mL/yr) vs the groups with asthma features (> or = 64 mL/yr) [p = 0.002]. Multivariable analysis showed that bronchodilator response, age, and smoking were significant predictors for FEV(1) decline but not asthma. CONCLUSIONS: Symptoms and signs of asthma are common in AAT deficiency and may start at the age of most rapid FEV(1) loss. Adjusting for other risk factors such as bronchodilator response, asthma as defined does not lead to an accelerated FEV(1) decline. In AAT deficiency, augmentation therapy is not more effective in preventing the loss of lung function in those with asthma compared to those without. PMID- 12628878 TI - Long-term repeatability of induced sputum cells and inflammatory markers in stable, moderately severe COPD. AB - STUDY OBJECTIVES: Neutrophilic inflammation is a major feature of COPD. Induced sputum is increasingly used to monitor inflammatory airway diseases. Although short-term repeatability of selected sputum markers has been extensively studied in several populations, data on the long-term repeatability of induced sputum markers in stable COPD are scant. DESIGN: Sputum supernatant of 12 patients with stable COPD was analyzed on three separate occasions with 4-weekly intervals. Sputum cells and inflammatory markers interleukin (IL)-8 and soluble intercellular adhesion molecule (sICAM)-1 were measured in supernatant using enzyme-linked immunosorbent assay. Repeatability of sputum markers was expressed by intraclass correlation coefficients (Ri). MEASUREMENTS AND RESULTS: Sputum induction was safe in all patients. None of the sputum parameters analyzed changed significantly throughout the study. The repeatability for cell differential counts in stable COPD was as follows: total cells, Ri = 0.07; neutrophils, Ri = 0.66; macrophages, Ri = 0.47; eosinophils, Ri = 0.49; and lymphocytes, Ri = 0.58. The repeatability of soluble markers was as follows: IL 8, Ri = 0.50; and sICAM, Ri = 0.58. Sputum neutrophils were negatively correlated with lung function on each separate occasion, whereas soluble markers were not correlated with sputum cells (p > 0.16, all correlations) or lung function (p > 0.24, all correlations). CONCLUSIONS: Clinically stable, moderate COPD is associated with equally stable sputum inflammatory markers. Repeatability of induced-sputum markers of neutrophilic inflammation in stable COPD is satisfactory, even over extended periods of time. These data support the usefulness of serial monitoring of induced-sputum inflammatory markers in COPD. PMID- 12628879 TI - Costs of chronic bronchitis and COPD: a 1-year follow-up study. AB - OBJECTIVE: This study attempted to determine the total direct costs derived from the management of chronic bronchitis and COPD in an ambulatory setting through a prospective, 1-year, follow-up study. METHOD: A total of 1,510 patients with chronic bronchitis and COPD were recruited from 268 general practices located throughout Spain. Patients were followed up for 1 year. All direct medical costs incurred by the cohort and related to their respiratory disease were quantified. Costs were calculated for patients with confirmed COPD according to the degree of severity of airflow obstruction. RESULTS: The global mean direct yearly cost of chronic bronchitis and COPD was $1,876. The cost generated by patients with COPD was $1,760, but the cost of severe COPD ($2,911) was almost double that of mild COPD ($1,484). Hospitalization costs represented 43.8% of costs, drug acquisition costs were 40.8%, and clinic visits and diagnostic tests represented only 15.4% of costs. CONCLUSION: This is the first prospective follow-up study on a large cohort of patients with chronic bronchitis and COPD aimed at quantifying direct medical costs under usual clinical practice in the community. Costs of chronic bronchitis and COPD were almost twofold those reported for asthma. Patterns of COPD management in the community differ from those recommended in guidelines. COPD represents a great health-care burden in developed countries, and aging of the population and continuing smoking habits predict that it will continue to do so in the future. PMID- 12628880 TI - Analysis of sequential aliquots of hypertonic saline solution-induced sputum from clinically stable patients with cystic fibrosis. AB - STUDY OBJECTIVES: Sputum induction (SI) is a noninvasive tool for sampling inflamed airways. The purpose of this study was to determine the optimal duration of collection in patients with cystic fibrosis (CF). The hypothesis was that the duration of SI collection would quantitatively and qualitatively alter the content of the induced sputum. METHODS: In 10 clinically stable patients with CF (mean +/- SD age, 28 +/- 7 years; mean FEV(1), 2.6 +/- 0.7 L), SI was performed with 3% hypertonic saline solution at five time points over 20 min. RESULTS: SI was well tolerated, with an average maximum fall in FEV(1) of 7 +/- 7%. The sample volumes, urea concentrations, interleukin-8 concentrations, total cell counts, and nonsquamous cell counts remained constant (p > 0.05). The percentage of neutrophils decreased from 89 +/- 5% to 86 +/- 4% (p = 0.03), and the percentage of alveolar macrophages increased 5 +/- 2% to 8 +/- 4% (p < 0.01). The mean quantitative microbiological counts of nonmucoid Pseudomonas aeruginosa and Staphylococcus aureus decreased over the 20-min time period each by half a log (p = 0.05 and p < 0.01, respectively). Surfactant protein-A concentration increased from 1.6 +/- 0.3 to 2.4 +/- 0.4 ng/mL (log(10); p < 0.001). CONCLUSIONS: We conclude that aliquots of induced sputum are similar in clinically stable patients with CF during 4-min intervals, although there is more alveolar sampling after 20 min. When induced-sputum samples are fractionated for research monitoring of inflammatory or microbiologic indexes, power calculations accounting for these variations over time are required. PMID- 12628881 TI - Aspergillus infection in lung transplant recipients with cystic fibrosis: risk factors and outcomes comparison to other types of transplant recipients. AB - STUDY OBJECTIVES: To characterize Aspergillus infections in lung transplant recipients with cystic fibrosis (CF). DESIGN: A retrospective analysis of 32 consecutive lung transplant recipients with CF who underwent bilateral lung transplant at the University of Wisconsin from 1994 to 2000 to determine the incidence, risk factors, and consequences of Aspergillus infection. The findings were compared to 101 non-CF recipients of lung transplants (93) and heart-lung transplants (8) for other transplant indications. SETTING: A university hospital. PATIENTS OR PARTICIPANTS: Lung transplant recipients with CF or other indications for transplantation. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Seventeen of 32 CF recipients (53%) had Aspergillus fumigatus isolated from their respiratory secretions prior to undergoing transplantation. Ten of these 17 (59%) recipients had A fumigatus persistently found in their respiratory secretions posttransplant vs 6 of 15 CF patients (40%) who had not been colonized pretransplant and 28 of 101 of the non-CF recipients (28%). Four of the preoperatively colonized CF recipients developed tracheobronchial aspergillosis (TBA) just distal to the bronchial anastomoses, and one recipient had dehiscence of the involved anastomosis. None of the CF recipients developed disseminated aspergillosis or pneumonia. Prophylactic antifungal therapy did not prevent TBA, and IV amphotericin B therapy was required to clear the infection in all four patients, with endobronchial debridement of necrotic tissue required in two of them. In contrast, 10 of the non-CF (10%) recipients developed Aspergillus infections posttransplant (TBA, 4 recipients; pneumonitis, 6 recipients), and only 3 patients had successful treatment and long-term survival (TBA, 2 patients; pneumonia, 1 patient). Donor lung ischemia time, cytomegalovirus infection or pneumonia, or pretransplant mechanical ventilation did not increase the risk of developing TBA in CF recipients. CONCLUSIONS: The risk of TBA for patients receiving lung transplants for CF warrants early surveillance bronchoscopy to detect TBA, particularly in recipients with pretransplant colonization. PMID- 12628882 TI - Thromboembolic disease involving the superior vena cava and brachiocephalic veins. AB - PURPOSE: To evaluate the frequency of diagnosis and the characteristics of brachiocephalic vein and superior vena cava (SVC) thromboembolic disease. BACKGROUND: Thromboembolic disease of the brachiocephalic veins or SVC rarely has been reported. In view of the frequent use of central venous access lines, it would seem that the percentage of hospitalized patients with thromboembolic disease of the brachiocephalic veins or SVC should be higher than is generally recognized. METHODS: A retrospective search for thromboembolic disease involving the brachiocephalic veins and SVC was made of patients who were hospitalized over a 2-year period. RESULTS: Thromboembolic disease of the brachiocephalic veins or SVC was diagnosed in 23 of 34,567 hospitalized adults (0.06%) who were > or = 20 years old. Two of 23 patients (8.7%) had pulmonary embolism. Cancer was present in 17 of 23 patients (74%), and 15 of 23 patients (65%) had central venous access lines. Edema of the arm, face, or neck was present in 21 of 23 patients (91%). Pain or discomfort was present 15 of 23 patients (65%). CONCLUSION: Isolated brachiocephalic vein and SVC thrombosis occur in a sufficient number of hospitalized patients to merit consideration of the diagnosis in patients who have cancer, central venous access lines, or both. The signs and symptoms of brachiocephalic vein thrombosis have features in common with SVC syndrome as well as with upper extremity deep venous thrombosis. In a patient with appropriate clinical findings, venography or other imaging may be indicated. PMID- 12628883 TI - Logistic regression analysis of potential prognostic factors for pulmonary thromboembolism. AB - OBJECTIVE: To identify potential prognostic factors for pulmonary thromboembolism (PTE), establishing a mathematical model to predict the risk for fatal PTE and nonfatal PTE. METHOD: The reports on 4,813 consecutive autopsies performed from 1979 to 1998 in a Brazilian tertiary referral medical school were reviewed for a retrospective study. From the medical records and autopsy reports of the 512 patients found with macroscopically and/or microscopically documented PTE, data on demographics, underlying diseases, and probable PTE site of origin were gathered and studied by multiple logistic regression. Thereafter, the "jackknife" method, a statistical cross-validation technique that uses the original study patients to validate a clinical prediction rule, was performed. RESULTS: The autopsy rate was 50.2%, and PTE prevalence was 10.6%. In 212 cases, PTE was the main cause of death (fatal PTE). The independent variables selected by the regression significance criteria that were more likely to be associated with fatal PTE were age (odds ratio [OR], 1.02; 95% confidence interval [CI], 1.00 to 1.03), trauma (OR, 8.5; 95% CI, 2.20 to 32.81), right-sided cardiac thrombi (OR, 1.96; 95% CI, 1.02 to 3.77), pelvic vein thrombi (OR, 3.46; 95% CI, 1.19 to 10.05); those most likely to be associated with nonfatal PTE were systemic arterial hypertension (OR, 0.51; 95% CI, 0.33 to 0.80), pneumonia (OR, 0.46; 95% CI, 0.30 to 0.71), and sepsis (OR, 0.16; 95% CI, 0.06 to 0.40). The results obtained from the application of the equation in the 512 cases studied using logistic regression analysis suggest the range in which logit p > 0.336 favors the occurrence of fatal PTE, logit p < - 1.142 favors nonfatal PTE, and logit P with intermediate values is not conclusive. The cross-validation prediction misclassification rate was 25.6%, meaning that the prediction equation correctly classified the majority of the cases (74.4%). CONCLUSIONS: Although the usefulness of this method in everyday medical practice needs to be confirmed by a prospective study, for the time being our results suggest that concerning prevention, diagnosis, and treatment of PTE, strict attention should be given to those patients presenting the variables that are significant in the logistic regression model. PMID- 12628884 TI - A randomized, phase III, double-blind, placebo-controlled trial of intrapleural instillation of methylprednisolone acetate in the management of malignant pleural effusion. AB - STUDY OBJECTIVES: To determine if intrapleural administration of methylprednisolone acetate (MA) after therapeutic thoracentesis for symptomatic malignant pleural effusion improved time to repeat thoracentesis for symptom control, quality of life (QOL), and dyspnea. DESIGN: Double-blind, randomized, placebo-controlled trial. SETTING: A tertiary care cancer treatment center in Edmonton, AB, Canada. PATIENT SELECTION: Patients with symptomatic pleural effusions secondary to disseminated malignancy requiring therapeutic thoracentesis for symptom control. INTERVENTIONS: Sixty-seven patients underwent ultrasound-guided therapeutic thoracentesis for management of symptomatic malignant pleural effusion. Patients were randomly and blindly assigned to either 160 mg (8 mL) of MA or 8 mL of saline solution instilled into the pleural space. Patients were followed up for 6 weeks to determine the time to repeat therapeutic thoracentesis. All patients completed the Functional Assessment of Cancer Therapy General (FACT-G) QOL questionnaire and a dyspnea visual analog scale (VAS) at baseline and again 2 weeks later. MEASUREMENTS AND RESULTS: Thirty-three patients received MA, and 34 patients received placebo; baseline characteristics for the two groups were similar, apart from a slightly higher use of concurrent systemic therapy in the placebo group. At 6 weeks follow-up, 50% of MA-treated patients required repeat thoracentesis compared to 56% of placebo-treated patients (not significant [NS]). The mean of the individual FACT-G change scores (2 weeks - baseline) was similar in the two groups (NS). VAS scores improved for both groups over the 2-week period, but the mean change scores (2 weeks - baseline) were not statistically different. CONCLUSION: Despite previous case series describing benefit from intrapleural MA in malignant pleural effusion, this controlled study of intrapleural MA instillation did not delay reaccumulation of symptomatic pleural effusion compared to placebo, nor were differences in QOL or dyspnea observed. PMID- 12628885 TI - The use of selective bronchography in predicting reversal of neoplastic obstructive atelectasis. AB - OBJECTIVES: To assess the ability of selective bronchography to predict which patients with neoplastic postobstructive atelectasis will respond to interventional therapies directed at the reexpansion of the affected lung. Furthermore, to compare the utility of selective bronchography with the current predictive standard that reversal of postobstructive atelectasis is unlikely when it is > or = 4 weeks in duration (ie, the 4-week rule). DESIGN: A prospective observational study. SETTING: A tertiary care referral center/medical school. PATIENTS: Twenty-seven consecutive patients with advanced lung cancer or other malignancy, with documented neoplastic postobstructive atelectasis involving a total of 44 lobes. INTERVENTIONS: Lobar collapse was documented radiographically. The duration of atelectasis was investigated and quantified as accurately as possible. Prior to the use of interventional therapies, selective bronchography was performed on each collapsed lobe, and the results were documented. Bronchography results did not influence the decision to proceed with interventional therapies. Patients had each of their collapsed lobes manipulated by interventional techniques that were directed at reexpansion of the lung. One week after the patient underwent the intervention, the degree of reexpansion was assessed radiographically. RESULTS: Interventional therapies leading to significant reversal of airway narrowing were completed in all 44 lobes. These were successful in reexpanding 28 of 44 collapsed lobes (64%). Selective bronchography demonstrated the following two distinct patterns: an intact bronchial tree (ie, tree pattern); or the absence of a distinguishable, distal bronchial tree (ie, blush pattern). The sensitivity of selective bronchography to predict reexpansion is 1.00 (95% confidence interval [CI], 0.90 to 1.00), and its specificity is 0.56 (95% CI, 0.30 to 0.80). There were no complications attributable to selective bronchography. The sensitivity of the 4-week rule to predict reexpansion is 0.61 (95% CI, 0.41 to 0.78), and its specificity is 0.75 (95% CI, 0.48 to 0.93). The results of selective bronchography and use of the 4 week rule were significantly different in predicting which lobes would reexpand and which would not (p = 0.0026). Using selective bronchography to predict the reversal of lobar atelectasis, the positive predictive value of the tree pattern was 0.80 and the negative predictive value of the blush pattern was 1.00. The values for the 4-week rule are 0.81 and 0.52, respectively. CONCLUSIONS: Selective bronchography is a useful tool for predicting whether patients with neoplastic postobstructive atelectasis would benefit from interventional techniques that are directed at lobar reexpansion. Selective bronchography appears to be superior to the 4-week rule in this regard. PMID- 12628887 TI - Long-term assessment of lung function in survivors of severe ARDS. AB - STUDY OBJECTIVES: To investigate the long-term outcome of lung function in survivors of severe ARDS after modern treatment strategies including lung protective mechanical ventilation and prone positioning maneuvers. DESIGN: Follow up cohort study. SETTING: University hospital pulmonary division and level 1 trauma center. PATIENTS: Sixteen survivors of severe ARDS (from 1992 to 1994) with a lung injury score > or = 2.5. MEASUREMENTS: The follow-up study (from 1995 to 1996) included interview, physical examination, chest radiographs, static and dynamic lung volumes, diffusion capacity of the lung for carbon monoxide (DLCO), blood gas analysis, and cardiopulmonary exercise testing (CPET). RESULTS: The mean +/- SD interval between hospital discharge and functional assessment was 29.5 +/- 8.7 months (range, 15.0 to 40.7 months). In approximately one half of the patients, mild abnormalities in static and dynamic lung volumes were found. In 25% (4 of 16 patients), lung function was obstructive; in 25% (4 of 16 patients), lung function was restrictive; and in 6.3% (1 of 16 patients), a combined obstructive-restrictive pattern was revealed. DLCO was impaired in 12.5% (2 of 16 patients); gas exchange during exercise was impaired in 45.5% (5 of 11 patients). CONCLUSIONS: Residual obstructive and restrictive defects as well as impaired pulmonary gas exchange remain common after severe ARDS. CPET is a very sensitive measure to evaluate residual impairment of lung function after ARDS. Using CPET, reduced pulmonary gas exchange can be detected in many patients with normal DLCO. PMID- 12628886 TI - Variability in antibiotic prescribing patterns and outcomes in patients with clinically suspected ventilator-associated pneumonia. AB - STUDY OBJECTIVES: To describe the variation in clinical practice strategies for the treatment of suspected ventilator-associated pneumonia (VAP) in a population of critically ill patients, and to determine whether initial empiric treatment with certain antibiotics, monotherapy vs combination antibiotic therapy, or appropriate vs inappropriate antibiotic therapy is associated with survival, length of hospital stay, or days free of antibiotics. DESIGN: Prospective, observational cohort study. SETTING: Medical-surgical ICUs of two university affiliated tertiary medical centers. PATIENTS: Between May 1, 1998, and August 1, 2000, we screened 7,030 ICU patients and identified 156 patients with clinically suspected VAP. Patients were followed up until death or discharge from the hospital. RESULTS: The mean age was 62 years, mean APACHE (acute physiology and chronic health evaluation) II score was 14, and mortality was 34%. Combination antibiotic therapy was used in 53% of patients. Piperacillin-tazobactam, fluoroquinolones, vancomycin, cephalosporins, and aminoglycosides were the most commonly employed antibiotics. Initial empiric antibiotics were deemed appropriate in 92% of patients. The predominant organisms isolated from respiratory secretions included Pseudomonas aeruginosa and Staphylococcus aureus. Patients had lower in-hospital mortality rates if their initial treatment regimen included an antipseudomonal penicillin plus beta-lactamase inhibitor (hazard ratio [HR], 0.41; 95% confidence interval [CI], 0.21 to 0.80; p = 0.009). There was also a strong trend toward reduced mortality rates in patients treated with aminoglycosides (HR, 0.43; 95% CI, 0.16 to 1.11; p = 0.08). Specific antibiotic therapy was not associated with length of hospital stay or days free of antibiotics. Outcomes were similar for patients treated with monotherapy vs combination therapy, and for patients who received initial appropriate vs inappropriate therapy. CONCLUSIONS: Patients with clinically suspected VAP who receive initial empiric therapy with antipseudomonal penicillins plus beta lactamase inhibitors, and possibly aminoglycosides, have lower in-hospital mortality rates when compared with those who are not treated with these antibiotics. These agents should be considered for the initial empiric therapy of VAP. PMID- 12628888 TI - Clinical utility of blood cultures drawn from central vein catheters and peripheral venipuncture in critically ill medical patients. AB - STUDY OBJECTIVE: To determine the sensitivity, specificity, and positive and negative predictive values of blood cultures obtained through a central vein catheter compared with peripheral venipuncture. DESIGN: Prospective cohort study. SETTING: A medical ICU (19 beds) from a university-affiliated urban teaching hospital. PATIENTS: Between February 2001 and October 2001, 300 paired blood culture specimens were obtained from 119 patients (2.52 paired cultures per patient). INTERVENTION: Prospective patient surveillance and data collection. MEASUREMENTS AND MAIN RESULTS: Thirty-four paired culture results (11.3%; 95% confidence interval, 7.8 to 14.8%) were accepted as true-positives representing a true bacteremia. The sensitivity of catheter-drawn and peripheral venipuncture samples was 82.4% and 64.7%, respectively, and specificity was 92.5% and 95.9%. The positive predictive value was 58.3% for catheter-drawn samples and 66.7% for peripheral venipuncture samples, and the respective negative predictive values were 97.6% and 95.5%. CONCLUSIONS: In critically ill medical patients, the negative predictive value of blood samples obtained by catheter draw or peripheral venipuncture for suspected bloodstream infection is good. However, the sensitivity of blood samples obtained by either catheter draw or peripheral venipuncture alone is not adequate to recommend the elimination of blood samples obtained from the other site. Clinicians should also be aware that additional blood samples may be necessary when interpreting positive blood culture results for common skin or central vein catheter contaminants. PMID- 12628889 TI - Pharmacologic effects of tobacco dust extract on isolated Guinea pig trachea. AB - STUDY OBJECTIVE: To determine the effects of tobacco dust extract (TDE) on isolated guinea pig tracheal smooth muscle. DESIGN: A controlled, in vitro smooth muscle study of the effect of pharmacologic agents on TDE. METHODS: The effect of TDE on isolated guinea pig tracheal smooth muscle was tested using water-soluble extracts of dust obtained from machines in a cigarette manufacturing plant. Dose related contractions of nonsensitized guinea pig trachea were demonstrated using these extracts. The dust extracts contained significant quantities of bacterial components (eg, endotoxin). Pharmacologic studies were performed by pretreating guinea pig tracheal tissue with drugs known to modulate smooth-muscle contraction: atropine, indomethacin, pyrilamine, nordihydroguaretic acid, acivicin, bromophenacyl bromide, 3,4,5-trimethoxybenzoic acid-8 (diethylamino)octyl ester, captopril, and capsaicin. RESULTS: Atropine strikingly reduced the contractile effects of these extracts. Inhibition of contraction by blocking other mediators was less complete. There was no inhibition of contraction by hexamethonium (10(-4) mol/L, 10(-5) mol/L, 10(-6) mol/L), suggesting that nicotine was not the major contractile mediator of TDE. A separate analysis using different molecular weight fractions of TDE indicated that the constrictor activity appears to be primarily in the fraction with a molecular weight < 10 kd. Additionally, the constrictor effect resided entirely in the nonlipid fraction of the extract. We suggest that TDE causes dose-related airway smooth-muscle constriction by nonimmunologic mechanisms involving a variety of airway mediators and possibly cholinergic receptors. CONCLUSIONS: The bronchoconstrictor activity of TDE resides primarily in its low molecular weight, nonlipid fraction, and hexamethonium studies suggest that this agent is not nicotine. PMID- 12628890 TI - Two aerosolized nitric oxide adducts as selective pulmonary vasodilators for acute pulmonary hypertension. AB - STUDY OBJECTIVES: To determine the selective vasodilatory effects of two inhaled "NONOate" aerosols in a closed chest pig model of acute pulmonary hypertension (APH). METHODS: APH was induced by IV infusion of the prostaglandin H(2)/thromboxane A(2) receptor agonist (U46619). Aerosolized diethylenetriamine nitric oxide (NO) adduct (DETA/NO, n = 4), dipropylenetriamine NO adduct (DPTA/NO, n = 4) [60 micro mol each], or placebo (n = 4) was delivered via the trachea. Hemodynamic parameters and blood samples were measured before and after inhalation therapy. RESULTS: Compared to control animals, pulmonary vascular resistance and pulmonary arterial pressure were significantly reduced from 10 to 105 min after DETA/NO administration and from 10 to 45 min after DPTA/NO aerosol administration (p < 0.05). Both aerosols had no significant effect on systemic vascular resistance or systemic BP. Serum nitrite significantly increased after the inhalation of both NONOates (p < 0.01). There was a tendency for reduced intrapulmonary shunting, particularly after treatment with DETA/NO. CONCLUSION: Both DETA/NO and DPTA/NO administered as aerosols selectively reduced pulmonary hypertension induced by U46619. PMID- 12628891 TI - Effects of chronic intermittent asphyxia on rat diaphragm and limb muscle contractility. AB - OBJECTIVE: In obstructive sleep apnea (OSA), there is intermittent upper airway (UA) collapse due to an imbalance between the collapsing force generated by the diaphragm and the stabilizing force of the UA muscles. This results in chronic intermittent asphyxia (CIA). We have previously shown that CIA affects UA muscle fatigue, but little is known about the effects of chronic hypoxia on diaphragm or on limb muscle contractile properties and structure. DESIGN: Rats were exposed to asphyxia and normoxia twice per minute for 8 h/d for 5 weeks to simulate the intermittent asphyxia of OSA in humans. Isometric contractile properties were determined from strips of isolated diaphragm, extensor digitorum longus (EDL), and soleus muscles in Krebs solution at 30 degrees C. EDL and soleus type 1 (slow, fatigue resistant), type 2A (fast, fatigue resistant), and type 2B (fast, fatigable) fiber distribution was determined using adenosine triphosphatase staining. RESULTS: CIA caused a significant increase in diaphragm, EDL, and soleus fatigue, and reduced recovery from fatigue. Most of the other contractile properties were unaffected aside from a small reduction in diaphragm half relaxation time and EDL twitch tension and a small shift to the left in the EDL force-frequency curve. There was no change in soleus fiber-type distribution and a small increase in EDL type 2A fibers (46.1 +/- 1.2% vs 49.9 +/- 1.4%, control vs CIA [mean +/- SD]). CONCLUSIONS: CIA increases diaphragm, EDL, and soleus muscle fatigue. We speculate that if this also occurs in OSA, it would contribute to the pathophysiology of the condition. PMID- 12628892 TI - Heliox vs air-oxygen mixtures for the treatment of patients with acute asthma: a systematic overview. AB - OBJECTIVE: To evaluate, by systematic review, the efficacy of heliox on respiratory mechanics and outcomes in patients with acute asthma. METHODS: The search strategy included searching electronic databases (MEDLINE, EMBASE, and The Cochrane Library) and the references of relevant articles. Study quality was assessed based on allocation concealment. Randomized controlled trials (RCTs) comparing heliox to an air-oxygen mixture (airO(2)) as an adjunct treatment in patients with acute asthmatic attacks were analyzed. For the qualitative portion of the analysis, all reports of the use of heliox in patients with acute asthma were included. RESULTS: Four RCTs (n = 278) were found to have a common respiratory parameter (peak expiratory flow rate as a percentage of predicted) suitable for meta-analysis. Within the 92% confidence interval (CI), there was a small benefit with the use of heliox compared to airO(2) (weighted mean difference, + 3%; 95% CI, - 2 to + 8%). There was also a slight improvement in the dyspnea index (weighted mean difference, 0.60; 95% CI, 0.04 to 1.16) with the use of heliox over airO(2). Overall, five RCTs, one nonrandomized unblinded parallel trial, one retrospective case-matched control trial, three case series, and one case report had results in favor of heliox; one RCT and one case series showed no improvement with heliox; one RCT showed a possible detrimental effect with heliox; and 1 small RCT was inconclusive. Most investigators did not prevent entrainment of room air during heliox use or compensate for the lower nebulizing efficiency of heliox. CONCLUSION: Based on surrogate markers, heliox may offer mild-to-moderate benefits in patients with acute asthma within the first hour of use, but its advantages become less apparent beyond 1 h, as most conventionally treated patients improve to similar levels, with or without it. The effect of heliox may be more pronounced in more severe cases. There are insufficient data on whether heliox can avert tracheal intubation, or change intensive care and hospital admission rates and duration, or mortality. PMID- 12628893 TI - Use of helium-oxygen mixtures in the treatment of acute asthma: a systematic review. AB - STUDY OBJECTIVE: To determine the effect of the addition of heliox to standard medical care on the course of acute asthma. DESIGN: Systematic review of randomized and nonrandomized prospective, controlled trials of children and adults that compared heliox to placebo when used in conjunction with other standard acute treatments. MAIN OUTCOME MEASURES: Pulmonary function tests, hospital admissions, physiologic measures, side effects, and clinical outcomes. RESULTS: Seven trials were selected for inclusion, with a total of 392 patients with acute asthma. Six studies involved adults, and one study dealt solely with children. The main outcome variable was spirometric measurements (peak expiratory flow or FEV(1)) in six trials. Two studies evaluated the effect of heliox on airways resistance. No significant differences were demonstrated between heliox or oxygen/air groups (standardized mean difference [SMD], - 0.20; 95% confidence interval [CI], - 0.91 to 0.51; p = 0.6). However, the four studies that used heliox to deliver nebulized therapy showed a nonsignificant increase in pulmonary function (SMD, - 0.21; 95% CI, - 0.43 to 0.01; p = 0.06). In two studies of the same subgroup, heliox mixtures produced a significantly greater increase of heart rate than oxygen/air (weighted mean difference, 9.0; 95% CI, 1.27 to 16.8; p = 0.02). However, the four studies that used heliox to deliver nebulized therapy reported a nonsignificant difference in hospital admissions (odds ratio, 1.07; 95% CI, 0.46 to 2.48; p = 0.9). Overall, heliox appears to be safe and well tolerated. CONCLUSIONS: The existing evidence does not provide support for the administration of helium-oxygen mixtures to emergency department patients with moderate-to-severe acute asthma. However, these conclusions are based on between group comparisons and small studies, and these results should be interpreted with caution. PMID- 12628894 TI - Adult toxicology in critical care: Part II: specific poisonings. PMID- 12628895 TI - An appraisal of multivariable logistic models in the pulmonary and critical care literature. AB - OBJECTIVE: Multivariable modeling techniques are appearing in today's medical literature with increasing frequency. Improper reporting of these statistical models can potentially make the results of a study inaccurate, misleading, or difficult to interpret. We performed a manual literature search of five international pulmonary and critical care journals to determine the accuracy in the reporting of logistic regression modeling strategies. DESIGN: We examined all of the published manuscripts for 12 potential limitations in the reporting of important statistical methodologies over a 6-month period from July 1, 2000, until December 31, 2000. RESULTS: Of the 81 articles that included multivariable logistic regression analyses, only 65% (53 analyses) properly reported the coding classification of pertinent independent variables that were included in the final model. An odds ratio and confidence interval were reported for the independent variables included in the final model for 79% (64 analyses) and 74% (60 analyses), respectively. Only 12% (10 articles) referenced whether interaction terms or effect modifications were examined, 1% (1 article) reported testing for collinearity, and only 16% (13 articles) included a goodness-of-fit analysis of the logistic model. The type of statistical package was reported in 69% (56 articles). Finally, approximately 39% of the articles (22 of 57) may have overfit the logistic regression model, leading to potentially unreliable regression coefficients and odds ratios. CONCLUSIONS: Our results indicate that the reporting of multivariable logistic regression analyses in the pulmonary and critical care literature is often incomplete, therefore making it difficult for the reader to accurately interpret the manuscript. We recommend the implementation of adequate guidelines that will lead to overall improvements in the reporting and possibly to the conducting of multivariable analyses in the pulmonary medicine and critical care medicine literature. PMID- 12628896 TI - Bilateral cavitary pulmonary consolidations in a patient undergoing allogeneic bone marrow transplantation for acute leukemia. PMID- 12628897 TI - Woman with dry cough and dyspnea on exertion has clubbing, conjunctival injection, and diffuse crackles. PMID- 12628898 TI - A cavitary lung lesion. PMID- 12628899 TI - Distraction therapy with nature sights and sounds reduces pain during flexible bronchoscopy: a complementary approach to routine analgesia. AB - STUDY OBJECTIVES: To determine whether distraction therapy with nature sights and sounds during flexible bronchoscopy (FB) reduces pain and anxiety. DESIGN: Randomized controlled trial. SETTING: Teaching hospital in Baltimore, MD. PATIENTS: Consecutive adult patients (n = 80) undergoing FB with conscious sedation. INTERVENTION: Nature scene murals were placed at the bedside, and patients were provided a tape of nature sounds to listen to before, during, and after the procedure. Patients assigned to the control group were not offered either the nature scene or the sounds. MEASUREMENTS AND RESULTS: The primary outcomes were patient ratings of pain control (a 5-point scale ranging from poor to excellent) and anxiety. In a multivariate ordinal logistic regression model, the odds of better pain control were greater in the intervention patients than in the control patients (odds ratio [OR], 4.76; 95% confidence interval [CI], 1.35 to 16.7), after adjustment for age, gender, race, education, health status, and dose of narcotic medication. Older patients and patients with better health status reported significantly less pain. There was no difference in patient reported anxiety between the two groups (OR, 0.87; 95% CI, 0.39 to 1.96). CONCLUSIONS: Distraction therapy with nature sights and sounds significantly reduces pain in patients undergoing FB. Although the precise mechanism of this beneficial effect requires further investigation, clinicians should consider this nonintrusive strategy in addition to standard analgesic medications in patients undergoing painful, invasive procedures. PMID- 12628900 TI - A novel form of manually assisted ventilation. AB - We report an individual with limb-girdle muscular dystrophy who has devised a way to assist her respiration by using her hands braced against the tray of her wheelchair. Utilizing this method, she was able to increase her tidal volume (VT) and lower her respiratory rate compared to unassisted spontaneous breathing, thereby maintaining a stable minute volume. The manually assisted VT measurements were comparable to those achieved using an intermittent abdominal pressure respirator (pneumatic belt). We believe that others with neuromuscular syndromes could use this technique, possibly decreasing their dependence on mechanical ventilatory assist devices. PMID- 12628901 TI - Pleural effusion caused by prostaglandin E1 preparation. AB - We encountered a case of bilateral pleural effusion associated with prostaglandin E(1) (PGE(1)) preparation. A 75-year-old man underwent replacement surgery for an amputated hand. PGE(1) was administered at 120 micro g/d to maintain circulation after vascularization. From day 7 of administration, respiratory distress developed. On day 12, pleural effusion was observed bilaterally. By discontinuing PGE(1) and improving hypoalbuminemia, pleural effusion resolved rapidly. This is the first case report of PGE(1)-induced pleural effusion; like other drug-induced pleural effusions, discontinuing the drug resulted in rapid improvement. Although a rare complication, pleural effusion has to be suspected when a patient receiving PGE(1) experiences difficulty with breathing. PMID- 12628902 TI - Multidrug-resistant tuberculosis in pregnancy: case report and review of the literature. AB - A woman at 23 weeks' gestation was treated with rifampin, isoniazid, and ethambutol for cavitary tuberculosis (TB). She did not respond within 3 weeks, and multidrug-resistant (MDR) TB was suspected. Direct plating on susceptibility media was performed immediately. Treatment was initiated with IV capreomycin, levofloxacin, para-aminosalicylic acid, pyrazinamide, cycloserine, and high-dose vitamin B(6) at 26 weeks' gestation. The patient delivered vaginally at week 35. The newborn was not infected. Following delivery, ethionamide was added as a sixth drug, and levofloxacin was replaced with moxifloxacin. The patient's sputum became smear-negative and culture-negative for TB. All reported cases of MDR-TB during pregnancy are reviewed. PMID- 12628903 TI - Pericardiocentesis and pancreatic aspiration needle biopsy in coagulopathic and thrombocytopenic cirrhotic patient. AB - We report on the case of a 40-year-old patient with coagulopathic alcoholic cirrhosis who underwent ultrasound-directed pericardiocentesis and fine-needle aspiration biopsy of the pancreas after receiving recombinant human factor VIIa (rhFVIIa). The infusion of rhFVIIa rapidly corrected her coagulopathy and made it possible to perform both procedures. The marked changes produced in the prothrombin time and international normalized ratio as a result of the infusion of rhFVIIa are presented. As a result of these changes in coagulation status, both procedures were performed safely, and the patient's clinical management and subsequent care plan were defined. PMID- 12628904 TI - COPD screening in high-risk groups. PMID- 12628905 TI - A difficult step in meta-analysis: refining the search. PMID- 12628906 TI - Sleep apnea devices and sleep apnea surgery should be compared on effectiveness, not efficacy. PMID- 12628907 TI - Orthotopic lung transplant for sarcoidosis. PMID- 12628908 TI - Lung cancer screening: contumacy vs mendacity. PMID- 12628909 TI - The best shot. PMID- 12628910 TI - Clarification of salmeterol studies. PMID- 12628911 TI - New diagnostic tool for tumor angiogenesis. PMID- 12628912 TI - Agenesis of pulmonary artery. PMID- 12628913 TI - Fatal liver injury associated with rifampin-pyrazinamide treatment of latent tuberculosis infection. PMID- 12628916 TI - Mrs2p is an essential component of the major electrophoretic Mg2+ influx system in mitochondria. AB - Steady-state concentrations of mitochondrial Mg(2+) previously have been shown to vary with the expression of Mrs2p, a component of the inner mitochondrial membrane with two transmembrane domains. While its structural and functional similarity to the bacterial Mg(2+) transport protein CorA suggested a role for Mrs2p in Mg(2+) influx into the organelle, other functions in cation homeostasis could not be excluded. Making use of the fluorescent dye mag-fura 2 to measure free Mg(2+) concentrations continuously, we describe here a high capacity, rapid Mg(2+) influx system in isolated yeast mitochondria, driven by the mitochondrial membrane potential Deltapsi and inhibited by cobalt(III)hexaammine. Overexpression of Mrs2p increases influx rates 5-fold, while the deletion of the MRS2 gene abolishes this high capacity Mg(2+) influx. Mg(2+) efflux from isolated mitochondria, observed with low Deltapsi only, also requires the presence of Mrs2p. Cross-linking experiments revealed the presence of Mrs2p-containing complexes in the mitochondrial membrane, probably constituting Mrs2p homo- oligomers. Taken together, these findings characterize Mrs2p as the first molecularly identified metal ion channel protein in the inner mitochondrial membrane. PMID- 12628917 TI - Plant mitochondria actively import DNA via the permeability transition pore complex. AB - Plant mitochondria are remarkable with respect to their content in foreign, alien and plasmid-like DNA, raising the question of the transfer of this information into the organelles. We demonstrate the existence of an active, transmembrane potential-dependent mechanism of DNA uptake into plant mitochondria. The process is restricted to double-strand DNA, but has no obvious sequence specificity. It is most efficient with linear fragments up to a few kilobase pairs. When containing appropriate information, imported sequences are transcribed within the organelles. The uptake likely involves the voltage-dependent anion channel and the adenine nucleotide translocator, i.e. the core components of the mitochondrial permeability transition pore complex in animal cells, but it does not rely on known mitochondrial membrane permeabilization processes. We conclude that DNA import into plant mitochondria might represent a physiological phenomenon with some functional relevance. PMID- 12628918 TI - Structure of a viral DNA gatekeeper at 10 A resolution by cryo-electron microscopy. AB - In tailed bacteriophages and herpes viruses, the viral DNA is packaged through the portal protein channel. Channel closure is essential to prevent DNA release after packaging. Here we present the connector structure from bacteriophage SPP1 using cryo-electron microscopy and single particle analysis. The multiprotein complex comprises the portal protein gp6 and the head completion proteins gp15 and gp16. Although we show that gp6 in the connector has a fold similar to that of the isolated portal protein, we observe conformational changes in the region of gp6 exposed to the DNA-packaging ATPase and to gp15. This reorganization does not cause closure of the channel. The connector channel traverses the full height of gp6 and gp15, but it is closed by gp16 at the bottom of the complex. Gp16 acts as a valve whose closure prevents DNA leakage, while its opening is required for DNA release upon interaction of the virus with its host. PMID- 12628920 TI - A ubiquitin-binding motif required for intramolecular monoubiquitylation, the CUE domain. AB - Monoubiquitylation is a regulatory signal, like phosphorylation, that can alter the activity, location or structure of a protein. Monoubiquitin signals are likely to be recognized by ubiquitin-binding proteins that transmit the regulatory information conferred by monoubiquitylation. To identify monoubiquitin binding proteins, we used a mutant ubiquitin that lacks the primary site of polyubiquitin chain formation as bait in a two-hybrid screen. The C-terminus of Vps9, a protein required in the yeast endocytic pathway, interacted specifically with monoubiquitin. The region required for monoubiquitin binding mapped to the Vps9 CUE domain, a sequence previously identified by database searches as similar to parts of the yeast Cue1 and mammalian Tollip proteins. We demonstrate that CUE domains bind directly to monoubiquitin and we have defined crucial interaction surfaces on both binding partners. The Vps9 CUE domain is required to promote monoubiquitylation of Vps9 by the Rsp5 hect domain ubiquitin ligase. Thus, we conclude that the CUE motif is an evolutionarily conserved monoubiquitin-binding domain that mediates intramolecular monoubiquitylation. PMID- 12628919 TI - Higher plant myosin XI moves processively on actin with 35 nm steps at high velocity. AB - High velocity cytoplasmic streaming is found in various plant cells from algae to angiosperms. We characterized mechanical and enzymatic properties of a higher plant myosin purified from tobacco bright yellow-2 cells, responsible for cytoplasmic streaming, having a 175 kDa heavy chain and calmodulin light chains. Sequence analysis shows it to be a class XI myosin and a dimer with six IQ motifs in the light chain-binding domains of each heavy chain. Electron microscopy confirmed these predictions. We measured its ATPase characteristics, in vitro motility and, using optical trap nanometry, forces and movement developed by individual myosin XI molecules. Single myosin XI molecules move processively along actin with 35 nm steps at 7 micro m/s, the fastest known processive motion. Processivity was confirmed by actin landing rate assays. Mean maximal force was approximately 0.5 pN, smaller than for myosin IIs. Dwell time analysis of beads carrying single myosin XI molecules fitted the ATPase kinetics, with ADP release being rate limiting. These results indicate that myosin XI is highly specialized for generation of fast processive movement with concomitantly low forces. PMID- 12628921 TI - A MAPK pathway mediates ethylene signaling in plants. AB - Ethylene signal transduction involves ETR1, a two-component histidine protein kinase receptor. ETR1 functions upstream of the negative regulator CTR1. The similarity of CTR1 to members of the Raf family of mitogen-activated protein kinase kinase kinases (MAPKKKs) suggested that ethylene signaling in plants involves a MAPK pathway, but no direct evidence for this has been provided. Here we show that distinct MAPKs are activated by the ethylene precursor aminocyclopropane-1-carboxylic acid (ACC) in Medicago and Arabidopsis: In Medicago, the ACC-activated MAPKs were SIMK and MMK3, while in Arabidopsis MPK6 and another MAPK were identified. Medicago SIMKK specifically mediated ACC induced activation of SIMK and MMK3. Transgenic Arabidopsis plants overexpressing SIMKK have constitutive MPK6 activation and ethylene-induced target gene expression. SIMKK overexpressor lines resemble ctr1 mutants in showing a triple response phenotype in the absence of ACC. Whereas MPK6 was not activated by ACC in etr1 mutants, ein2 and ein3 mutants showed normal activation profiles. In contrast, ctr1 mutants showed constitutive activation of MPK6. These data indicate that a MAPK cascade is part of the ethylene signal transduction pathway in plants. PMID- 12628922 TI - p53 induction and activation of DDR1 kinase counteract p53-mediated apoptosis and influence p53 regulation through a positive feedback loop. AB - DDR1, discoidin domain receptor 1, belongs to a subfamily of tyrosine kinase receptors with an extracellular domain homologous to Dictyostellium discoideum protein discoidin 1. We showed that DDR1 is a direct p53 transcriptional target, and that DNA damage induced a p53-dependent DDR1 response associated with activation of its tyrosine kinase. We further demonstrated that DDR1 activated the MAPK cascade in a Ras-dependent manner. Whereas levels of p53, phosphoserine 15 p53, p21, ARF and Bcl-X(L) were increased in response to exogenous overexpression of activated DDR1, dominant-negative DDR1 inhibited irradiation induced MAPK activation and p53, phosphoserine-15 p53, as well as induced p21 and DDR1 levels, suggesting that DDR1 functions in a feedforward loop to increase p53 levels and at least some of its effectors. Nonetheless, inhibition of DDR1 function resulted in strikingly increased apoptosis of wild-type p53-containing cells in response to genotoxic stress through a caspase-dependent pathway. These results strongly imply that this p53 response gene must predominately act to alleviate the adverse effects of stress induced by p53 on its target cell. PMID- 12628923 TI - Protein kinase CK2 and protein kinase D are associated with the COP9 signalosome. AB - The COP9 signalosome (CSN) purified from human erythrocytes possesses kinase activity that phosphoryl ates proteins such as c-Jun and p53 with consequence for their ubiquitin (Ub)-dependent degradation. Here we show that protein kinase CK2 (CK2) and protein kinase D (PKD) co-purify with CSN. Immunoprecipitation and far western blots reveal that CK2 and PKD are in fact associated with CSN. As indicated by electron microscopy with gold-labeled ATP, at least 10% of CSN particles are associated with kinases. Kinase activity, most likely due to CK2 and PKD, co-immuno precipitates with CSN from HeLa cells. CK2 binds to DeltaCSN3(111-403) and CSN7, whereas PKD interacts with full-length CSN3. CK2 phosphorylates CSN2 and CSN7, and PKD modifies CSN7. Both CK2 and PKD phosphorylate c-Jun as well as p53. CK2 phosphoryl ates Thr155, which targets p53 to degradation by the Ub system. Curcumin, emodin, DRB and resveratrol block CSN associated kinases and induce degradation of c-Jun in HeLa cells. Curcumin treatment results in elevated amounts of c-Jun-Ub conjugates. We conclude that CK2 and PKD are recruited by CSN in order to regulate Ub conjugate formation. PMID- 12628924 TI - Transcriptional activation of the NF-kappaB p65 subunit by mitogen- and stress activated protein kinase-1 (MSK1). AB - Nuclear factor kappaB (NF-kappaB) is one of the key regulators of transcription of a variety of genes involved in immune and inflammatory responses. NF-kappaB activity has long been thought to be regulated mainly by IkappaB family members, which keep the transcription factor complex in an inactive form in the cytoplasm by masking the nuclear localization signal. Nowadays, the importance of additional mechanisms controlling the nuclear transcription potential of NF kappaB is generally accepted. We show that the mitogen-activated protein kinase inhibitors SB203580 and PD98059 or U0126, as well as a potent mitogen- and stress activated protein kinase-1 (MSK1) inhibitor H89, counteract tumor necrosis factor (TNF)-mediated stimulation of p65 transactivation capacity. Mutational analysis of p65 revealed Ser276 as a target for phosphorylation and transactivation in response to TNF. Moreover, we identified MSK1 as a nuclear kinase for p65, since MSK1 associates with p65 in a stimulus-dependent way and phosphorylates p65 at Ser276. This effect represents, together with phosphorylation of nucleosome components such as histone H3, an essential step leading to selective transcriptional activation of NF-kappaB-dependent gene expression. PMID- 12628925 TI - GRIM-19, a death-regulatory gene product, suppresses Stat3 activity via functional interaction. AB - Signal transducer and activator of transcription 3 (Stat3) is a latent cytoplasmic transcription factor that can be activated by cytokines and growth factors. Stat3 plays important roles in cell growth, anti-apoptosis and cell transformation, and is constitutively active in various cancers. We examined its potential regulators by yeast two-hybrid screening. GRIM-19, a gene product related to interferon-beta- and retinoic acid-induced cancer cell death, was identified and demonstrated to interact with Stat3 in various cell types. The interaction is specific for Stat3, but not for Stat1 and Stat5a. The interaction regions in both proteins were mapped, and the cellular localization of the interaction was examined. GRIM-19 itself co-localizes with mitochondrial markers, and forms aggregates at the perinulear region with co-expressed Stat3, which inhibits Stat3 nuclear translocation stimulated by epidermal growth factor (EGF). GRIM-19 represses Stat3 transcriptional activity and its target gene expression, and also suppresses cell growth in Src-transformed cells and a Stat3-expressing cell line. Our data suggest that GRIM-19 is a novel negative regulator of Stat3. PMID- 12628926 TI - Purification and functional characterization of the human N-CoR complex: the roles of HDAC3, TBL1 and TBLR1. AB - Corepressors N-CoR and SMRT participate in diverse repression pathways and exist in large protein complexes including HDAC3, TBL1 and TBLR1. However, the roles of these proteins in SMRT-N-CoR complex function are largely unknown. Here we report the purification and functional characterization of the human N-CoR complex. The purified N-CoR complex contains 10-12 associated proteins, including previously identified components and a novel actin-binding protein IR10. We show that TBL1/TBLR1 associates with N-CoR through two independent interactions: the N terminal region and the C-terminal WD-40 repeats interact with the N-CoR RD1 and RD4 region, respectively. In vitro, TBL1/TBLR1 bind histones H2B and H4, and, importantly, repression by TBL1/TBLR1 correlates with their interaction with histones. By using specific small interference RNAs (siRNAs), we demonstrate that HDAC3 is essential, whereas TBL1 and TBLR1 are functionally redundant but essential for repression by unliganded thyroid hormone receptor. Together, our data reveal the roles of HDAC3 and TBL/TBLR1 and provide evidence for the functional importance of histone interaction in repression mediated by SMRT-N-CoR complexes. PMID- 12628927 TI - Transcription factor YY1 functions as a PcG protein in vivo. AB - Polycomb group (PcG) proteins function as high molecular weight complexes that maintain transcriptional repression patterns during embryogenesis. The vertebrate DNA binding protein and transcriptional repressor, YY1, shows sequence homology with the Drosophila PcG protein, pleiohomeotic (PHO). YY1 might therefore be a vertebrate PcG protein. We used Drosophila embryo and larval/imaginal disc transcriptional repression systems to determine whether YY1 repressed transcription in a manner consistent with PcG function in vivo. YY1 repressed transcription in Drosophila, and this repression was stable on a PcG-responsive promoter, but not on a PcG-non-responsive promoter. PcG mutants ablated YY1 repression, and YY1 could substitute for PHO in repressing transcription in wing imaginal discs. YY1 functionally compensated for loss of PHO in pho mutant flies and partially corrected mutant phenotypes. Taken together, these results indicate that YY1 functions as a PcG protein. Finally, we found that YY1, as well as Polycomb, required the co-repressor protein CtBP for repression in vivo. These results provide a mechanism for recruitment of vertebrate PcG complexes to DNA and demonstrate new functions for YY1. PMID- 12628928 TI - A novel splicing regulator shares a nuclear import pathway with SR proteins. AB - Alternative splicing of precursor mRNA is often regulated by serine/arginine-rich proteins (SR proteins) and hnRNPs, and varying their concentration in the nucleus can be a mechanism for controlling splice site selection. To understand the nucleocytoplasmic transport mechanism of splicing regulators is of key importance. SR proteins are delivered to the nucleus by transportin-SRs (TRN SRs), importin beta-like nuclear transporters. Here we identify and characterize a non-SR protein, RNA-binding motif protein 4 (RBM4), as a novel substrate of TRN SR2. TRN-SR2 interacts specifically with RBM4 in a Ran-sensitive manner. TRN-SR2 indeed mediates the nuclear import of a recombinant protein containing the RBM4 C terminal domain. This domain serves as a signal for both nuclear import and export, and for nuclear speckle targeting. Finally, both in vivo and in vitro splicing analyses demonstrate that RBM4 not only modulates alternative pre-mRNA splicing but also acts antagonistically to authentic SR proteins in splice site and exon selection. Thus, a novel splicing regulator with opposite activities to SR proteins shares an identical import pathway with SR proteins to the nucleus. PMID- 12628930 TI - In vivo transposition mediated by V(D)J recombinase in human T lymphocytes. AB - The rearrangement of immunoglobulin (Ig) and T-cell receptor (TCR) genes in lymphocytes by V(D)J recombinase is essential for immunological diversity in humans. These DNA rearrangements involve cleavage by the RAG1 and RAG2 (RAG1/2) recombinase enzymes at recombination signal sequences (RSS). This reaction generates two products, cleaved signal ends and coding ends. Coding ends are ligated by non-homologous end-joining proteins to form a functional Ig or TCR gene product, while the signal ends form a signal joint. In vitro studies have demonstrated that RAG1/2 are capable of mediating the transposition of cleaved signal ends into non-specific sites of a target DNA molecule. However, to date, in vivo transposition of signal ends has not been demonstrated. We present evidence of in vivo inter-chromosomal transposition in humans mediated by V(D)J recombinase. T-cell isolates were shown to contain TCRalpha signal ends from chromosome 14 inserted into the X-linked hypo xanthine-guanine phosphoribosyl transferase locus, resulting in gene inactivation. These findings implicate V(D)J recombinase-mediated transposition as a mutagenic mechanism capable of deleterious genetic rearrangements in humans. PMID- 12628929 TI - The path from nucleolar 90S to cytoplasmic 40S pre-ribosomes. AB - Recent reports have increased our knowledge of the consecutive steps during 60S ribosome biogenesis substantially, but 40S subunit formation is less well understood. Here, we investigate the maturation of nucleolar 90S pre-ribosomes into cytoplasmic 40S pre-ribosomes. During the transition from 90S to 40S particles, the majority of non-ribosomal proteins (approximately 30 species) dissociate, and significantly fewer factors associate with 40S pre-ribosomes. Notably, some of these components are part of both early 90S and intermediate 40S pre-particles in the nucleolus (e.g. Enp1p, Dim1p and Rrp12p), whereas others (e.g. Rio2p and Nob1p) are found mainly on late cytoplasmic pre-40S subunits. Finally, temperature-sensitive mutants mapping either in earlier (enp1-1) or later (rio2-1) components exhibit defects in the formation and nuclear export of pre-40S subunits. Our data provide an initial biochemical map of the pre-40S ribosomal subunit on its path from the nucleolus to the cytoplasm. This pathway involves fewer changes in composition than seen during 60S biogenesis. PMID- 12628931 TI - High-resolution sperm typing of meiotic recombination in the mouse MHC Ebeta gene. AB - Meiotic crossovers detected by pedigree analysis in the mouse MHC cluster into hotspots. To explore the properties of hotspots, we subjected the class II E(beta) gene to high-resolution sperm crossover analysis. We confirm the presence of a highly localized hotspot 1.0-1.6 kb wide in the second intron of E(beta) and show that it is flanked by DNA which is almost completely recombinationally inert. Mice heterozygous for haplotype s and another MHC haplotype show major haplotype-dependant variation in crossover rate but always the same hotspot, even in crosses including the highly diverged p haplotype. Crossovers in reciprocal orientations occur at similar rates but show different distributions across the hotspot, with the position of centre points in the two orientations shifted on average by 400 bp. This asymmetry results in crossover products showing biased gene conversion in favour of hotspot markers from the non-initiating haplotype, and supports the double-strand break repair model of recombination, with haplotype s as the most efficient crossover initiator. The detailed behaviour of the E(beta) hotspot, including evidence for highly localized recombination initiation, is strikingly similar to human hotspots. PMID- 12628932 TI - The complex between a four-way DNA junction and T7 endonuclease I. AB - The junction-resolving enzyme endonuclease I is selective for the structure of the DNA four-way (Holliday) junction. The enzyme binds to a four-way junction in two possible orientations, with a 4:1 ratio, opening the DNA structure at the centre and changing the global structure into a 90 degrees cross of approximately coaxial helices. The nuclease cleaves the continuous strands of the junction in each orientation. Binding leads to pronounced regions of protection of the DNA against hydroxyl radical attack. Using all this information together with the known structure of the enzyme and the structure of the BglI-DNA complex, we have constructed a model of the complex of endonuclease I and a DNA junction. This shows how the enzyme is selective for the structure of a four-way junction, such that both continuous strands can be accommodated into the two active sites so that a productive resolution event is possible. PMID- 12628933 TI - Protein motion from non-specific to specific DNA by three-dimensional routes aided by supercoiling. AB - DNA-binding proteins are generally thought to locate their target sites by first associating with the DNA at random and then translocating to the specific site by one-dimensional (1D) diffusion along the DNA. We report here that non-specific DNA conveys proteins to their target sites just as well when held near the target by catenation as when co-linear with the target. Hence, contrary to the prevalent view, proteins move from random to specific sites primarily by three-dimensional (3D) rather than 1D pathways, by multiple dissociation/re-association events within a single DNA molecule. We also uncover a role for DNA supercoiling in target-site location. Proteins find their sites more readily in supercoiled than in relaxed DNA, again indicating 3D rather than 1D routes. PMID- 12628934 TI - Pathway utilization in response to a site-specific DNA double-strand break in fission yeast. AB - We have examined the genetic requirements for efficient repair of a site-specific DNA double-strand break (DSB) in Schizosaccharomyces pombe. Tech nology was developed in which a unique DSB could be generated in a non-essential minichromosome, Ch(16), using the Saccharomyces cerevisiae HO-endonuclease and its target site, MATa. DSB repair in this context was predominantly through interchromosomal gene conversion. We found that the homologous recombination (HR) genes rhp51(+), rad22A(+), rad32(+) and the nucleotide excision repair gene rad16(+) were required for efficient interchromosomal gene conversion. Further, DSB-induced cell cycle delay and efficient HR required the DNA integrity checkpoint gene rad3(+). Rhp55 was required for interchromosomal gene conversion; however, an alternative DSB repair mechanism was used in an rhp55Delta background involving ku70(+) and rhp51(+). Surprisingly, DSB-induced minichromosome loss was significantly reduced in ku70Delta and lig4Delta non-homologous end joining (NHEJ) mutant backgrounds compared with wild type. Furthermore, roles for Ku70 and Lig4 were identified in suppressing DSB-induced chromosomal rearrangements associated with gene conversion. These findings are consistent with both competitive and cooperative interactions between components of the HR and NHEJ pathways. PMID- 12628937 TI - Antibiotic treatment of atherosclerotic cardiovascular disease. PMID- 12628936 TI - Mdm2 haplo-insufficiency profoundly inhibits Myc-induced lymphomagenesis. AB - Mdm2 harnesses the p53 tumor suppressor, yet loss of one Mdm2 allele in Mdm2(+/-) mice has heretofore not been shown to impair tumor development. Here we report that Mdm2 haplo-insufficiency profoundly suppresses lymphomagenesis in E micro myc transgenic mice. Mdm2(+/-)E micro -myc transgenics had greatly protracted rates of B cell lymphoma development with life spans twice that of wild-type transgenic littermates. Im paired lymphoma development was associated with drastic reductions in peripheral B cell numbers in Mdm2(+/-)E micro -myc transgenics, and primary pre-B cells from Mdm2(+/-)E micro -myc transgenics and Mdm2(+/-) littermates were extremely susceptible to spontaneous apoptosis. Loss of p53 rescued all of the effects of Mdm2 haplo-insufficiency, indicating they were p53 dependent. Furthermore, half of the lymphomas that ultimately emerged in Mdm2(+/-)E micro -myc transgenics harbored inactivating mutations in p53, and the majority overcame haplo-insufficiency by overexpressing Mdm2. These results support the concept that Mdm2 functions are rate limiting in lymphomagenesis and that targeting Mdm2 will enhance p53-mediated apoptosis, compromising tumor development and/or maintenance. PMID- 12628935 TI - Protein kinase Cdelta is responsible for constitutive and DNA damage-induced phosphorylation of Rad9. AB - The mammalian homolog of the Schizosaccharomyces pombe Rad9 is involved in checkpoint signaling and the induction of apoptosis. While the mechanisms responsible for the regulation of human Rad9 (hRad9) are not known, hRad9 is subject to hyperphosphorylation in the response of cells to DNA damage. The present results demonstrate that protein kinase Cdelta (PKCdelta) associates with Rad9 and that DNA damage induces this interaction. PKCdelta phosphorylates hRad9 in vitro and in cells exposed to genotoxic agents. The functional significance of the interaction between hRad9 and PKCdelta is supported by the finding that activation of PKCdelta is necessary for formation of the Rad9-Hus1-Rad1 complex. We also show that PKCdelta is required for binding of hRad9 to Bcl-2. In concert with these results, inhibition of PKCdelta attenuates Rad9-mediated apoptosis. These findings demonstrate that PKCdelta is responsible for the regulation of Rad9 in the Hus1-Rad1 complex and in the apoptotic response to DNA damage. PMID- 12628938 TI - Use of biomarkers in the management of heart failure: are we there yet? PMID- 12628939 TI - Drug therapy and heart failure prevention. PMID- 12628940 TI - Allograft arteriosclerosis and immune-driven angiogenesis. PMID- 12628941 TI - Prosthetic heart valves and pregnancy. PMID- 12628942 TI - Bone marrow-derived cardiomyocytes are present in adult human heart: A study of gender-mismatched bone marrow transplantation patients. AB - BACKGROUND: Recent studies have identified cardiomyocytes of extracardiac origin in transplanted human hearts, but the exact origin of these myocyte progenitors is currently unknown. METHODS AND RESULTS: Hearts of female subjects (n=4) who had undergone sex-mismatched bone marrow transplantation (BMT) were recovered at autopsy and analyzed for the presence of Y chromosome-positive cardiomyocytes. Four female gender-matched BMT subjects served as controls. Fluorescence in situ hybridization (FISH) for the Y chromosome was performed on paraffin-embedded sections to identify cells of bone marrow origin with concomitant immunofluorescent labeling for alpha-sarcomeric actin to identify cardiomyocytes. A total of 160 000 cardiomyocyte nuclei were analyzed approximating 20 000 nuclei per patient. The mean percentage of Y chromosome-positive cardiomyocytes in patients with sex-mismatched BMT was 0.23+/-0.06%. Not a single Y chromosome positive cardiomyocyte was identified in any of the control patients. Immunofluorescent costaining for laminin and chromosomal ploidy analysis with FISH showed no evidence of either pseudonuclei or cell fusion in any of the chimeric cardiac myocytes identified. CONCLUSIONS: These data establish for the first time human bone marrow as a source of extracardiac progenitor cells capable of de novo cardiomyocyte formation. PMID- 12628943 TI - Randomized study comparing radiofrequency ablation with cryoablation for the treatment of atrial flutter with emphasis on pain perception. AB - BACKGROUND: Radiofrequency ablation (RF) of atrial flutter (AFL) has a high procedural efficacy, a low recurrence rate, and reports of procedure-related pain. The aim of the present study was to compare RF with cryoablation (cryo) for the treatment of AFL, with emphasis on pain perception during application of energy. METHODS AND RESULTS: Fourteen patients (55+/-11 years, 11 males) with AFL were randomized to receive ablation of the cavotricuspid isthmus (CTI) by either RF or cryo. Cryothermia was delivered with the CryoCor Cryoablation System (10F, 6-mm tip), and radiofrequency energy was delivered with the use of an 8-mm-tip catheter. Pain was evaluated according to a visual analogue scale (VAS; 0 to 100). All patients in the cryo group were successfully ablated with a mean of 18 applications (9 sites), and RF was successful in 6 of 7 patients (not significant) with 13 applications (not significant). The mean temperature was -82 degrees C and 55 degrees C for cryo and RF, respectively. One patient in the cryo group perceived pain, versus all 7 patients in the RF group (P<0.05). The proportion of painful applications averaged 75.3% in the RF group and 2.0% in the cryo group (P<0.05), whereas the corresponding VAS for pain was 38.3+/-25.3 and 0.32+/-0.86, respectively (P<0.05). At 6-month follow-up, there were no recurrences of atrial flutter. CONCLUSION: Cryo, as compared with RF, produces significantly less pain during application. Although in the present study there was no significant difference in efficacy, larger studies will be needed to definitively compare efficacy. PMID- 12628945 TI - Is the association between parity and coronary heart disease due to biological effects of pregnancy or adverse lifestyle risk factors associated with child rearing? Findings from the British Women's Heart and Health Study and the British Regional Heart Study. AB - BACKGROUND: Parity is associated with coronary heart disease (CHD) risk. In the present study, we assessed the associations between number of children and CHD in both women and men. METHODS AND RESULTS: A total of 4286 women and 4252 men aged 60 to 79 years from 24 British towns were studied. Number of children was positively associated with body mass index and waist-hip ratio in both sexes. In women but not in men, number of children was inversely associated with high density lipoprotein cholesterol and was positively associated with triglycerides and diabetes. For both sexes, similar "J" shaped associations between number of children and CHD were observed, with the prevalence lowest among those with 2 children and increasing linearly with each additional child beyond 2. For those with at least 2 children, each additional child increased the age-adjusted odds of CHD by 30% (odds ratio, 1.30; 95% confidence interval, 1.17 to 1.44) for women and by 12% for men (odds ratio, 1.12; 95% confidence interval, 1.02 to 1.22). Adjustment for obesity and metabolic risk factors attenuated the associations between greater number of children and CHD in both sexes, although in women some association remained. CONCLUSIONS: Lifestyle risk factors associated with child rearing lead to obesity and result in increased CHD in both sexes; biological responses of pregnancy may have additional adverse effects in women. PMID- 12628944 TI - Antibiotic therapy after acute myocardial infarction: a prospective randomized study. AB - BACKGROUND: Infection with Chlamydia pneumoniae is suspected to contribute to the pathogenesis of human atherosclerosis. We investigated whether treatment with the macrolide antibiotic roxithromycin would reduce mortality or morbidity in patients with an acute myocardial infarction. METHODS AND RESULTS: Eight hundred seventy-two patients with an acute myocardial infarction (AMI) were randomly assigned to receive double-blind treatment with either 300 mg roxithromycin or placebo daily for 6 weeks. Primary end point was total mortality during 12-month follow-up. Four hundred thirty-three patients were treated with roxithromycin and 439 with placebo. With the exception of a higher proportion of patients suffering an anterior wall AMI (48.1% in the roxithromycin group versus 40.2% in the placebo group; P=0.027) and a lower prevalence of chronic obstructive pulmonary disease in the roxithromycin group (3.5% versus 6.9%, P=0.028), baseline characteristics, reperfusion therapy, and medical treatment were well balanced between the two groups. More patients in the roxithromycin group interrupted their study medication before completion of at least 4 weeks of treatment (78 of 433 [18%] versus 48 of 439 [11%]; P=0.003; odds ratio, 1.8; 95% CI, 1.2 to 2.6). Follow-up at 12 months was achieved in 868 of 872 (99.5%) patients. Total mortality at 12 months was 6.5% (28 of 431) in the roxithromycin group compared with 6.0% (26 of 437) in the placebo group (odds ratio, 1.1; 95% CI, 0.6 to 1.9; P=0.739). There were also no differences in the secondary combined end points at 12 months. CONCLUSIONS: Treatment of AMI patients with roxithromycin did not reduce event rates during 12 months of follow-up. Therefore, our findings do not support the routine use of antibiotic treatment with a macrolide in patients with AMI. PMID- 12628946 TI - Heparin-coated stent placement for the treatment of stenoses in small coronary arteries of symptomatic patients. AB - BACKGROUND: The role of stents, especially of heparin-coated stents for the treatment of stenoses in small coronary arteries, is still unclear. Therefore, we performed this prospective, randomized trial to evaluate the angiographic and clinical outcome after treatment of stenoses in small coronary arteries (2.0 to 2.6 mm) of symptomatic patients. METHODS AND RESULTS: We randomly assigned 588 patients to angioplasty (n=195), bare stenting (n=196), or heparin-coated stenting (n=197). The primary end point was minimal lumen diameter (MLD) at 6 months. With comparable baseline parameters, the two stent arms showed a larger postinterventional MLD, larger acute gain, and smaller residual percent diameter stenosis, although a residual stenosis of 12+/-16% was achieved in the angioplasty arm, including a 27% crossover rate to stenting. Eighty percent of patients had follow-up angiography, which documented a borderline significantly larger MLD and smaller percent diameter stenosis for the two stent groups (1.34+/ 0.48 mm and 42+/-20% after angioplasty, 1.47+/-0.48 mm and 36+/-20% after bare stenting, and 1.45+/-0.54 mm and 38+/-23% after heparin-coated stenting; P=0.049 and P=0.038, respectively), but restenosis rates were not different (32%, 25%, and 30%). Thrombotic events occurred in 1.0% after angioplasty and 0.5% after bare or heparin-coated stenting. Survival without myocardial infarction or target vessel revascularization at 250 days was 84.6% (angioplasty), 88.3% (bare stenting), and 88.3% (heparin-coated stenting; log-rank P=0.39). CONCLUSION: Compared with angioplasty with provisional stenting, bare and heparin-coated stenting confer superior angiographic results and a nonsignificant 24% reduction in clinical events, with no difference between bare and heparin-coated stenting in the treatment of stenoses in small coronary arteries. PMID- 12628947 TI - Twenty-year survival after coronary artery surgery: an institutional perspective from Emory University. AB - BACKGROUND: Coronary artery bypass graft (CABG) surgery has been performed frequently for symptomatic coronary atherosclerotic heart disease for more than 30 years. However, uncertainty exists regarding the relationship between long term survival after CABG and readily available clinical correlates of mortality. METHODS AND RESULTS: We studied outcome at 20 years by age, sex, and other variables in 3939 patients who had CABG surgery from 1973 to 1979 in the Emory University System of Healthcare. Twenty-year survival, freedom from myocardial infarction, and freedom from repeat CABG were 35.6% (95% confidence interval [CI], 33.9% to 37.3%), 66.6% (95% CI, 64.6% to 68.6%), and 59.1% (95% CI, 56.9% to 61.5%). Multivariate correlates of late mortality were age (hazard ratio [HR], 1.46 per 10 years), female sex (HR, 1.21), hypertension (HR, 1.44), angina class (HR, 1.07 per class increase of 1), prior CABG (HR, 1.72), ejection fraction (HR, 1.07 per 10-point decrease), number of vessels diseased (HR, 1.11 per 1-vessel increase), and weight (HR, 1.04 per 10 kg). Twenty-year survival by age was 55%, 38%, 22%, and 11% for age <50, 50 to 59, 60 to 69, and >70 years at the time of initial surgery. Survival at 20 years after surgery with and without hypertension was 27% and 41%, respectively. Similarly, 20-year survival was 37% and 29% for men and women. CONCLUSIONS: Symptomatic coronary atherosclerotic heart disease requiring surgical revascularization is progressive with continuing events and mortality. Clinical correlates of mortality significantly impact survival over time and may help identify long-term benefits after CABG. PMID- 12628948 TI - Changes in brain natriuretic peptide and norepinephrine over time and mortality and morbidity in the Valsartan Heart Failure Trial (Val-HeFT). AB - BACKGROUND: Neurohormones are considered markers of heart failure progression. We examined whether changes in brain natriuretic peptide (BNP) and norepinephrine (NE) over time are associated with corresponding changes in mortality and morbidity in the Valsartan Heart Failure Trial. METHODS AND RESULTS: Plasma BNP and NE were measured before randomization and during follow-up in approximately 4300 patients in the Valsartan Heart Failure Trial. The relation between baseline BNP and NE and all-cause mortality and first morbid event (M&M) was analyzed in subgroups, with values above and below the median, and by quartiles. The change and percent change from baseline to 4 and 12 months in BNP and NE were also analyzed by quartiles for subsequent M&M. Risk ratios for M&M were calculated using a Cox proportional hazard model. Risk ratio of M&M for patients with baseline BNP or NE above the median was significantly higher than that for patients with values below the median. Baseline BNP and NE in quartiles also showed a quartile-dependent increase in M&M. BNP had a stronger association with M&M than NE. Patients with the greatest percent decrease in BNP and NE from baseline to 4 and 12 months had the lowest whereas patients with greatest percent increase in BNP and NE had the highest M&M. CONCLUSIONS: Not only are plasma BNP and NE important predictors of heart failure M&M, but changes in these neurohormones over time are associated with corresponding changes in M&M. These data further reinforce their role as significant surrogate markers in HF and underscore the importance of including their measurement in HF trials. PMID- 12628949 TI - Prevention of Heart Failure in Patients in the Heart Outcomes Prevention Evaluation (HOPE) Study. AB - BACKGROUND: Previous trials in the prevention of heart failure have been restricted to patients with low ejection fraction or hypertension. We assessed an angiotensin-converting enzyme (ACE) inhibitor, ramipril, to prevent the development of heart failure in high-risk patients without known low ejection fraction or heart failure. METHODS AND RESULTS: We randomly assigned 9297 patients to receive double-blind ramipril (10 mg daily) or matching placebo for 4.5 years. Death attributable to heart failure, hospitalization for heart failure, initiation of open-label ACE inhibitor for heart failure, or development of typical signs or symptoms of heart failure developed in 951 patients and was associated with a 4.01-fold increase in the risk of death (P<0.0001). The rate of developing heart failure was significantly increased with coronary disease (risk ratio, 2.17), microalbuminuria (1.82), left ventricular hypertrophy (1.47), increasing age (by decade, 1.37), and diabetes (1.36). Ramipril reduced new-onset heart failure rate from 11.5% to 9.0% (relative risk, 0.77; 95% CI, 0.68 to 0.87; P<0.0001). Ramipril consistently reduced heart failure rate both in those with (relative risk, 0.87) and those without an interim myocardial infarction (relative risk, 0.78). Ramipril also reduced the heart failure rate more in patients with baseline systolic pressure above the median (139 mm Hg) (relative risk, 0.67) compared with those below the median (relative risk, 0.91; P=0.024 for interaction of group by treatment). CONCLUSION: Ramipril significantly reduces the rate of development of heart failure in patients at high risk of cardiovascular events. PMID- 12628950 TI - Enalapril reduces the incidence of diabetes in patients with chronic heart failure: insight from the Studies Of Left Ventricular Dysfunction (SOLVD). AB - BACKGROUND: Diabetes mellitus is a predictor of morbidity and mortality in patients with heart failure. The effect of angiotensin-converting enzyme (ACE) inhibitors on the prevention of diabetes in patients with left ventricular dysfunction is unknown. The aim of this retrospective study was to assess the effect of the ACE inhibitor enalapril on the incidence of diabetes in the group of patients from the Montreal Heart Institute enrolled in the Studies of Left Ventricular Dysfunction (SOLVD). METHODS AND RESULTS: Clinical charts were evaluated for fasting plasma glucose (FPG) levels by blinded reviewers. A diagnosis of diabetes was made when a FPG > or =126 mg/dL (7 mmol/L) was found at 2 visits (follow-up, 2.9+/-1.0 years). Of the 391 patients enrolled at the Montreal Heart Institute, 291 were not diabetic (FPG <126 mg/dL without a history of diabetes), 153 of these were on enalapril and 138 were on placebo. Baseline characteristics were similar in the 2 groups. Forty patients developed diabetes during follow-up, 9 (5.9%) in the enalapril group and 31 (22.4%) in the placebo group (P<0.0001). By multivariate analysis, enalapril remained the most powerful predictor for risk reduction of developing diabetes (hazard ratio, 0.22; 95% confidence intervals, 0.10 to 0.46; P<0.0001). The effect of enalapril was striking in the subgroup of patients with impaired FPG (110 mg/dL [6.1 mmol/L] < or =FPG <126 mg/dL) at baseline: 1 patient (3.3%) in the enalapril group versus 12 (48.0%) in the placebo group developed diabetes (P<0.0001). CONCLUSIONS: Enalapril significantly reduces the incidence of diabetes in patients with left ventricular dysfunction, especially those with impaired FPG. PMID- 12628951 TI - Isolated ambulatory hypertension predicts cardiovascular morbidity in elderly men. AB - BACKGROUND: Little is known about isolated ambulatory hypertension, a state with elevated ambulatory but normal office blood pressure (BP). This study aimed to investigate the prognostic significance of isolated ambulatory hypertension for cardiovascular morbidity in a population of elderly men. METHODS AND RESULTS: At baseline, 24-hour ambulatory BP and metabolic and cardiac risk profiles were evaluated in 578 untreated 70-year-old men, participants of a population-based cohort. Subjects with isolated ambulatory hypertension (office BP <140/90 and daytime BP > or =135/85) and sustained hypertension (office BP > or =140/90 and daytime BP > or =135/85) had increased plasma glucose, body mass index, and echocardiographically determined left ventricular relative wall thickness compared with normotensive subjects (office BP <140/90 and daytime BP <135/85). Seventy-two cardiovascular morbid events (2.37 per 100 person-years at risk) occurred over 8.4 years of follow-up. The prognostic value of isolated ambulatory and sustained hypertension was assessed with Cox proportional hazard regression. Multivariate models adjusting for serum cholesterol, smoking, and diabetes demonstrated that both isolated ambulatory hypertension (hazard ratio [HR], 2.77; 95% CI, 1.15 to 6.68) and sustained hypertension (HR, 2.94; 95% CI, 1.49 to 5.82) were independent predictors of cardiovascular morbidity. In a multivariate model with continuous BP variables, ambulatory daytime systolic BP (HR for 1 SD increase, 1.47; 95% CI, 1.09 to 1.97) was associated with an adverse outcome independently of office systolic BP. CONCLUSIONS: In the present study, isolated ambulatory hypertension as well as sustained hypertension predicted cardiovascular morbidity. The findings suggest that 24-hour ambulatory BP monitoring may disclose important prognostic information also in subjects characterized as normotensive according to office BP. PMID- 12628952 TI - Cardiovascular morbidity and mortality in women diagnosed with rheumatoid arthritis. AB - BACKGROUND: Rheumatoid arthritis may be associated with an increased risk of cardiovascular disease. We compared the incidence rates of myocardial infarction and stroke in subjects with and without rheumatoid arthritis. METHODS AND RESULTS: A prospective cohort study was conducted among the 114 342 women participating in the Nurses' Health Study who were free of cardiovascular disease and rheumatoid arthritis at baseline in 1976. All self-reported cases of rheumatoid arthritis were confirmed by medical record review. Fatal and nonfatal myocardial infarctions and strokes were similarly confirmed. Multivariate pooled logistic regression was used to adjust for potential cardiovascular risk factors. Five hundred twenty-seven incident cases of rheumatoid arthritis and 3622 myocardial infarctions and strokes were confirmed during 2.4 million person-years of follow-up. The adjusted relative risk of myocardial infarction in women with rheumatoid arthritis compared with those without was 2.0 (95% confidence interval [CI], 1.23 to 3.29). For stroke, the adjusted relative risk was 1.48 (95% CI, 0.70 to 3.12). Women who had rheumatoid arthritis for at least 10 years had a risk for myocardial infarction of 3.10 (95% CI, 1.64 to 5.87). CONCLUSION: In this large prospective cohort of women, participants with rheumatoid arthritis had a significantly increased risk of myocardial infarction but not stroke compared with those without rheumatoid arthritis. If these data are confirmed, aggressive coronary heart disease prevention strategies should be tested for persons with rheumatoid arthritis. PMID- 12628953 TI - Angiopoietin-1 protects against the development of cardiac allograft arteriosclerosis. AB - BACKGROUND: Angiopoietin (Ang)-1 is an angiogenic growth factor that counteracts the permeability and proinflammatory effects of vascular endothelial growth factor and other proinflammatory cytokines. Recently, we demonstrated that vascular endothelial growth factor enhances cardiac allograft arteriosclerosis. Here, we studied the roles of Ang1, its natural antagonist Ang2, and their receptor Tie2 in rat cardiac allograft arteriosclerosis. METHODS AND RESULTS: Heterotopic cardiac allografts and syngrafts were transplanted from Dark Agouti (DA) to Wistar-Furth rats and from DA to DA rats, respectively. Immunohistochemistry disclosed that only a few mesenchymal cells expressed Ang1 in normal hearts and syngrafts, whereas no immunoreactivity was found in cardiac allografts undergoing chronic rejection. Ang2 and Tie2 immunoreactivity was induced mainly in capillaries and postcapillary venules in chronic allografts when compared with syngeneic controls, but no immunoreactivity was found in arterial endothelium. Intracoronary perfusion of cardiac allografts with a clinical-grade adenoviral vector encoding human Ang1 (Ad.Ang1) protected against the development of allograft arteriosclerosis. Ad.Ang1 perfusion reduced Ang2 expression in microcirculation, the numbers of graft-infiltrating leukocytes, and the level of immunoactivation and interstitial fibrosis, as well as both the incidence and intensity of intimal lesions. Ad.Ang1 perfusion also increased CD34+ stem cell counts in peripheral blood. CONCLUSIONS: Our findings suggest that the antiinflammatory properties of Ang1 may offer an entirely new therapeutic approach to prevent cardiac allograft arteriosclerosis. PMID- 12628954 TI - Eliminating atherogenesis in mice by switching off hepatic lipoprotein secretion. AB - BACKGROUND: LDL receptor-deficient "apolipoprotein (apo)-B100-only" mice (Ldlr-/ Apob100/100 have elevated LDL cholesterol levels on a chow diet and develop severe aortic atherosclerosis. We hypothesized that both the hypercholesterolemia and the susceptibility to atherosclerosis could be eliminated by switching off hepatic lipoprotein production. METHODS AND RESULTS: We bred Ldlr-/-Apob100/100 mice that were homozygous for a conditional allele for Mttp (the gene for microsomal triglyceride transfer protein) and the inducible Mx1-Cre transgene. In these animals, which we called "Reversa mice," the hypercholesterolemia could be reversed, without modifying the diet or initiating a hypolipidemic drug, by the transient induction of Cre expression in the liver. After Cre induction, hepatic Mttp expression was virtually eliminated (as judged by quantitative real-time PCR), hepatic lipoprotein secretion was abolished (as judged by electron microscopy), and LDLs were virtually eliminated from the plasma. Intestinal lipoprotein production was unaffected. In mice fed a chow diet, Cre induction reduced plasma cholesterol levels from 233.9+/-46.0 to 37.2+/-6.5 mg/dL. In mice fed a high-fat diet, cholesterol levels fell from 525.7+/-32.2 to 100.6+/-14.3 mg/dL. The elimination of hepatic lipoprotein production completely prevented both the development of atherosclerosis and the changes in gene expression that accompany atherogenesis. CONCLUSIONS: We developed mice in which hypercholesterolemia can be reversed with a genetic switch. These mice will be useful for understanding gene-expression changes that accompany the reversal of hypercholesterolemia and atherosclerosis. PMID- 12628955 TI - Stromal cell-derived factor-1 effects on ex vivo expanded endothelial progenitor cell recruitment for ischemic neovascularization. AB - BACKGROUND: Stromal cell-derived factor-1 (SDF-1) is a chemokine considered to play an important role in the trafficking of hematopoietic stem cells. Given the close relationship between hematopoietic stem cells and endothelial progenitor cells (EPCs), we investigated the effect of SDF-1 on EPC-mediated vasculogenesis. METHODS AND RESULTS: Flow cytometric analysis demonstrated expression of CXCR4, the receptor of SDF-1, by 66+/-3% of EPCs after 7 days in culture. In vitro modified Boyden chamber assay showed a dose-dependent EPC migration toward SDF-1 (control versus 10 ng/mL SDF-1 versus 100 ng/mL SDF-1, 24+/-2 versus 71+/-3 versus 140+/-6 cells/mm2; P<0.0001). SDF-1 attenuated EPC apoptosis (control versus SDF-1, 27+/-1 versus 7+/-1%; P<0.0001). To investigate the effect of SDF-1 in vivo, we locally injected SDF-1 into athymic ischemic hindlimb muscle of nude mice combined with human EPC transplantation to determine whether SDF-1 augmented EPC-induced vasculogenesis. Fluorescence microscopic examination disclosed increased local accumulation of fluorescence-labeled EPCs in ischemic muscle in the SDF-1 treatment group (control versus SDF-1=241+/-25 versus 445+/-24 cells/mm2, P<0.0001). At day 28 after treatment, ischemic tissue perfusion was improved in the SDF-1 group and capillary density was also increased. (control versus SDF-1, 355+/-26 versus 551+/-30 cells/mm2; P<0.0001). CONCLUSION: These findings indicate that locally delivered SDF-1 augments vasculogenesis and subsequently contributes to ischemic neovascularization in vivo by augmenting EPC recruitment in ischemic tissues. PMID- 12628956 TI - Bosentan for the prevention of overcirculation-induced experimental pulmonary arterial hypertension. AB - BACKGROUND: The dual endothelin-receptor antagonist bosentan has been reported to improve pulmonary arterial hypertension, but the role of endothelins in the pathogenesis of the condition remains uncertain. We investigated the roles of endothelin-1 (ET-1), nitric oxide (NO), vascular endothelial growth factor (VEGF), and tenascin in overcirculation-induced pulmonary hypertension in piglets, as a model of early pulmonary arterial hypertension, with or without bosentan therapy. METHODS AND RESULTS: Thirty 3-week-old piglets were randomized to placebo or to bosentan 15 mg/kg BID after the anastomosis of the left subclavian artery to the pulmonary arterial trunk or after a sham operation. Three months later, the animals underwent a hemodynamic evaluation followed by cardiac and pulmonary tissue sampling for morphometry, immunohistochemistry, and real-time quantitative PCR. Chronic systemic-to-pulmonary shunting increased circulating plasma ET-1, pulmonary mRNA for ET-1, ET(B) receptor, inducible NO synthase, VEGF, and pulmonary ET-1 and VEGF proteins. There were increases in myocardial mRNA for ET(A) receptor and VEGF and in myocardial VEGF protein. Pulmonary and myocardial tissue mRNA for tenascin did not change. Normalized-flow pulmonary artery pressure increased from 20 (2) to 33 (1) mm Hg [mean (SEM)], arteriolar medial thickness increased on average by 83%, and these changes were completely prevented by bosentan therapy. Right ventricular end-systolic elastance increased in proportion to pulmonary arterial elastance with or without bosentan. CONCLUSIONS: Experimental overcirculation-induced pulmonary arterial hypertension appears to be causally related to an activation of the pulmonary ET 1 system and as such is completely prevented by the dual endothelin receptor antagonist bosentan. PMID- 12628957 TI - Review of the International Position Paper on Women's Health and Menopause: a comprehensive approach. PMID- 12628958 TI - Images in cardiovascular medicine. Sirolimus-eluting stent implanted in human coronary artery for 16 months: pathological findings. PMID- 12628959 TI - Images in cardiovascular medicine. Spontaneous onset of atrioventricular nodal reentrant tachycardia with high grade intranodal block. PMID- 12628960 TI - Prognosis of aortic intramural hematoma with and without penetrating atherosclerotic ulcer: a clinical and radiological analysis. PMID- 12628961 TI - Multiple plaque rupture in acute coronary syndrome. PMID- 12628962 TI - Multiple atherosclerotic plaque rupture in acute coronary syndrome. PMID- 12628964 TI - Chimerism as a mechanism of self-repair. PMID- 12628963 TI - Long-term use of contraceptive depot medroxyprogesterone acetate in young women impairs arterial endothelial function assessed by cardiovascular magnetic resonance. PMID- 12628965 TI - Combining composite endpoints: counterintuitive or a mathematical impossibility? PMID- 12628967 TI - Transgenic modeling of interleukin-13 in the lung. AB - Interleukin (IL)-13 is a key cytokine in asthma pathogenesis. We used constitutive and inducible overexpression transgenic mice to characterize the mechanisms by which IL-13 causes phenotypic alterations in the lung. These studies demonstrated that chemokine receptor-2, transforming growth factor beta(1), and IL-11 play an important role in the regulation of inflammation and remodeling in the IL-13-treated lung. The study results also demonstrated that IL 13 induces vascular endothelial growth factor, which causes bronchial circulation neovascularization in the murine airway. Last, it was demonstrated that IL-13 induces adenosine accumulation and that adenosine in turn stimulates IL-13 elaboration. These approaches validated in vivo genetic targets against which therapies can be directed to selectively regulate aspects of the IL-13 phenotype. PMID- 12628968 TI - Introduction: asthma in the new millennium. PMID- 12628969 TI - Isolation and characterization of hemopoietic cells from lungs of allergic mice. AB - We developed a procedure for the isolation of hemopoietic cells from murine lung. Ovalbumin sensitization and challenge increased the numbers of functionally intact hemopoietic progenitors recovered from digested lung fragments by 80-fold to 120-fold, relative to naive controls. Eosinophil precursors, which are absent in the naive mouse lung, accumulated in the lungs of sensitized/challenged mice. Progenitors in allergic BALB/c mice were recoverable from lung parenchyma, not blood or airways, and were exclusively CD34+. Precursors isolated from allergic lung, unlike those from bone marrow, were inhibited by dexamethasone and were stimulated by prostaglandin D(2). This directly demonstrates that sensitized/challenged lungs accumulate hemopoietic progenitors and precursors, distinct from those in bone marrow. PMID- 12628970 TI - Overlap between death receptor and non-receptor-mediated mechanisms during apoptosis in human eosinophils. PMID- 12628971 TI - Contribution of the distal lung to the pathologic and physiologic changes in asthma: potential therapeutic target Roger S. Mitchell lecture. AB - Pathologic and physiologic evidence has emerged in the last few years suggesting that the airway inflammation and remodeling that characterize asthma occur not only in the central airways but extend to the distal lung and the lung parenchyma. The distal airways are capable of producing T helper (Th) type 2 cytokines and chemokines, and, more recently, they have been recognized as a predominant site of airflow obstruction in asthmatic patients. In the lung parenchyma, a similar Th2 cytokine profile and infiltration of inflammatory cells also has been reported. The inflammation at this distal site has been described as being more severe when compared to the large amount of airway inflammation, and evidence of remodeling in the lung periphery is emerging. The recognition of asthma as a disease of the entire respiratory tract has an important clinical significance highlighting the need to also consider the distal lung as a target in any therapeutic strategy for effective treatment of this disease. PMID- 12628972 TI - Hydrolysis of surfactant phospholipids catalyzed by phospholipase A2 and eosinophil lysophospholipases causes surfactant dysfunction: a mechanism for small airway closure in asthma. PMID- 12628973 TI - The functional consequences of structural changes in the airways: implications for airway hyperresponsiveness in asthma. PMID- 12628974 TI - Interleukin-4 and interleukin-13 augment the proliferative effect of dexamethasone on distal lung fibroblasts. PMID- 12628975 TI - Role of fibrin in determining airway closure. PMID- 12628976 TI - Mapping susceptibility genes for allergic diseases. AB - Allergic diseases are most likely due to interactions between genetic and environmental factors. While many of the environmental components have been studied for years, only recently has significant progress been made in identifying the genes responsible for susceptibility or expression of these diseases. Genome-wide screens in various populations have identified the locations of susceptibility genes for asthma and atopy, as well as associated phenotypes such as bronchial hyperresponsiveness and increased total serum IgE levels. In addition, this positional cloning approach has led to the discovery of several genes for asthma or related phenotypes, which is extending our understanding of the pathophysiology of allergic diseases. As these genes are identified and characterized, the relationships of these genes to each other and the environment will become important areas of research. Understanding the basic interactions that lead to the development of allergy and asthma will lead to new therapeutic approaches that will be used to modify the development and clinical progression of these common disorders. PMID- 12628977 TI - Peripheral lung responsiveness assessed by forced oscillations through the wedged bronchoscope. PMID- 12628978 TI - Effect of dexamethasone on beta2-adrenergic desensitization in airway smooth muscle: role of the ARG19 polymorphism. PMID- 12628979 TI - Macrophage inflammatory protein-2 levels are associated with changes in serum leptin concentrations following ozone-induced airway inflammation. PMID- 12628980 TI - The genetics of innate immunity in the lung. PMID- 12628981 TI - Increased messenger RNA expression of c-maf and GATA-3 after segmental allergen challenge in allergic asthmatics. PMID- 12628982 TI - Increased expression of interleukin-9 messenger RNA after segmental allergen challenge in allergic asthmatics. PMID- 12628983 TI - UCB 62045: pharmacology of a novel, dual-function anti-inflammatory agent with histamine type 1 receptor antagonist and 5-lipoxygenase inhibitor activity. PMID- 12628984 TI - The relationship of induced-sputum inflammatory cells to BAL and biopsy. PMID- 12628985 TI - Interferon-gamma reduces interleukin-4- and interleukin-13-augmented transforming growth factor-beta2 production in human bronchial epithelial cells by targeting Smads. PMID- 12628986 TI - Eosinophil cationic protein concentration in saliva does not correlate with eosinophil cationic protein concentration in sputum. PMID- 12628987 TI - Interleukin-13 alters mucociliary differentiation of human nasal epithelial cells. PMID- 12628988 TI - Lack of association between the tumor necrosis factor-alpha regulatory region genetic polymorphisms associated with elevated tumor necrosis factor-alpha levels and children with asthma. PMID- 12628989 TI - Identification of genes and proteins regulated by interleukin-5 in human eosinophils using microarrays and two-dimensional electrophoresis/mass spectrometry. PMID- 12628990 TI - Interleukin-13 induces surfactant function abnormality in the murine lung. PMID- 12628991 TI - Beta2-adrenergic receptor polymorphisms in children with status asthmaticus. PMID- 12628993 TI - Developing the epithelial, viral, and allergic paradigm for asthma: Giles F. Filley lecture. PMID- 12628992 TI - Human eosinophil group IID secretory phospholipase A2 causes surfactant dysfunction. PMID- 12628994 TI - Modulation of vasodilator-stimulated phosphoprotein in vivo in human epithelial cells by segmental allergen challenge and beta2-agonist therapy. PMID- 12628995 TI - Proliferation of the airway epithelium in asthma: are inflammatory cells required? PMID- 12628997 TI - Histamine decreases E-cadherin-based adhesion to increase permeability of human airway epithelium. PMID- 12628996 TI - Is asthma an infectious disease?: Thomas A. Neff lecture. AB - Respiratory tract infections caused by viruses, Chlamydia, and Mycoplasma have been implicated in the pathogenesis of asthma. Of these respiratory pathogens, viruses have been demonstrated to be associated with asthma epidemiologically in at least two ways. First, during infancy, certain viruses have been implicated as potentially being responsible for the inception of the asthmatic phenotype. Second, in patients with established asthma, particularly children, viral upper respiratory tract infections play a significant role in producing acute exacerbations of airway obstruction that may result in frequent outpatient visits or in hospitalizations. For infections with other microbial agents, recent attention has focused on Chlamydia and Mycoplasma as potential contributors to both exacerbations and the severity of chronic asthma in terms of loss of lung function or medication requirements. In an attempt to address the question posed in the title, this article will briefly review these various associations as they pertain to the pathogenesis of asthma in both children and adults. PMID- 12628998 TI - Hygiene hypothesis of asthma: a murine asthma model with Mycoplasma pneumoniae infection. PMID- 12628999 TI - Mechanisms that potentially underlie virus-induced exaggerated inflammatory responses by airway epithelial cells. PMID- 12629000 TI - Actin dynamics: a potential integrator of smooth muscle (Dys-)function and contractile apparatus gene expression in asthma. Parker B. Francis lecture. PMID- 12629001 TI - Concentration of cytokines and growth factors in BAL fluid after allergen challenge in asthmatics and their effect on alpha-smooth muscle actin and collagen III synthesis by human lung fibroblasts. PMID- 12629003 TI - Severe/fatal asthma. AB - Severe asthma is poorly understood clinically, physiologically, and pathologically. While milder forms of asthma are generally easily treated, more severe forms often remain refractory to the best current medical care. Although some patients with severe asthma have had severe disease for most of their lives, there appears to be a second group that develops severe disease in adulthood. Additionally, it is not clear which genetic and environmental elements may be the most important in the development of severe disease. Physiologically, these patients often have airtrapping and may have loss of elastic recoil, as well. The pathology demonstrates a heterogeneity of findings, including continued eosinophilic inflammation, structural changes, distal disease, and, in at least one third of patients, a different pathology. Treatment remains problematic and likely will remain so until a better understanding of this disease develops. PMID- 12629002 TI - Nocturnal asthma. AB - Lung function in a healthy individual varies in a circadian rhythm, with peak lung function occurring near 4:00 PM (1600 hours) and minimal lung function occurring near 4:00 AM (0400 hours). An episode of nocturnal asthma is characterized by an exaggeration in this normal variation in lung function from daytime to nighttime, with diurnal changes in pulmonary function generally of > 15%. The occurrence of nocturnal asthma is associated with increased morbidity and inadequate asthma control, and has an important negative impact on quality of life (QOL). Newer data have shed light on physiologic and immunologic mechanisms that underlie the nocturnal development of airway obstruction. It remains controversial whether nocturnal asthma is a distinct entity or is a manifestation of more severe asthma. The current data do not resolve these two alternatives, as well-controlled studies have reached opposite conclusions. However, the clinical associations of gastroesophageal reflux disease and obesity appear to be strong. The treatment of asthma with effective controller agents can reduce nighttime symptoms, improve psychometric outcomes, and improve QOL. PMID- 12629004 TI - Hypothalamic-pituitary-adrenal axis dysfunction during sleep in nocturnal asthma. PMID- 12629005 TI - Allergen-specific early neutrophil infiltration after allergen challenge in a murine model. PMID- 12629006 TI - Airway hyperresponsiveness. AB - Airway hyperresponsiveness is a characteristic feature of asthma and consists of an increased sensitivity of the airways to an inhaled constrictor agonist, a steeper slope of the dose-response curve, and a greater maximal response to the agonist. Measurements of airway responsiveness are useful in making a diagnosis of asthma, particularly in patients who have symptoms that are consistent with asthma and who have no evidence of airflow obstruction. These tests can be performed quickly, safely, and reproducibly. Certain inhaled stimuli, such as environmental allergens, increase airway inflammation and enhance airway hyperresponsiveness. These changes in airway hyperresponsiveness are of much smaller magnitude than those seen when asthmatic patients with persistent airway hyperresponsiveness are compared to healthy subjects. They are, however, similar to changes occurring in asthmatic patients that are associated with worsening asthma control. The mechanisms of the transient allergen-induced airway hyperresponsiveness are not likely to fully explain the underlying mechanisms of the persistent airway hyperresponsiveness in asthmatic patients. PMID- 12629007 TI - Intercellular adhesion molecule-1 plays a pivotal role in endotoxin-induced airway disease. PMID- 12629009 TI - Airway remodeling in asthma. AB - Chronic inflammation and remodeling may follow acute inflammation or may begin insidiously as a low-grade smoldering response, especially in the case of immune reactions. The histologic hallmarks of chronic inflammation and remodeling are as follows: (1) infiltration by macrophages and lymphocytes; (2) proliferation of fibroblasts that may take the form of myofibroblasts; (3) angiogenesis; (4) increased connective tissue (fibrosis); and (5) tissue destruction. It is clear that changes in the extracellular matrix, smooth muscle, and mucous glands have the capacity to influence airway function and reactivity in asthma patients. However, it is not known how each of the many structural changes that occur in the airway wall contributes to altered airway function in asthma. In asthma, remodeling is almost always present in biopsy specimens (eg, collagen deposition on basement membrane) but is not always clinically demonstrated. Destruction and subsequent remodeling of the normal bronchial architecture are manifested by an accelerated decline in FEV(1) and bronchial hyperresponsiveness. This irreversible component of airway obstruction is more prominent in patients with severe disease and even persists after aggressive anti-inflammatory treatment. Airway remodeling appears to be of great importance for understanding the long term follow-up of asthmatic patients, but there are major gaps in our knowledge. Physiologic correlations with pathology represent a major missing link that should be filled. More long-term studies are needed to appreciate the prevention and treatment of remodeling. Future research therefore should provide better methods for limiting airway remodeling in asthma patients. PMID- 12629008 TI - Provocation with adenosine 5'-monophosphate increases sputum eosinophils. PMID- 12629010 TI - Interleukin-13 stimulates the proliferation of lung myofibroblasts via a signal transducer and activator of transcription-6-dependent mechanism: a possible mechanism for the development of airway fibrosis in asthma. PMID- 12629011 TI - Interleukin-4 promotes airway remodeling in asthma: regulation of procollagen I (alpha1) gene by interleukin-4. PMID- 12629013 TI - Pulmonary surfactant proteins A and D recognize lipid ligands on Mycoplasma pneumoniae and markedly augment the innate immune response to the organism. PMID- 12629012 TI - Mycoplasma pneumoniae antigens stimulate interleukin-8. PMID- 12629014 TI - Lipopolysaccharide down-regulates leukotriene C4 synthase gene expression in a cell-specific manner in the monocyte-like cell line, THP-1. PMID- 12629015 TI - Synergetic effect of interleukin-4 and transforming growth factor-beta1 on type I collagen gel contraction and degradation by HFL-1 cells: implication in tissue remodeling. PMID- 12629016 TI - Effects of inhaled fluticasone on bronchial hyperresponsiveness and airway inflammation in Mycoplasma pneumoniae-infected mice. PMID- 12629018 TI - Increased expression of platelet-derived growth factor receptor-beta in airway fibroblasts of severe asthmatics. PMID- 12629017 TI - Quantitative assessment of subepithelial collagen deposition in the airways of mice following ovalbumin sensitization and intratracheal challenge. PMID- 12629019 TI - Modulation of fibroblast growth factor expression and signal transduction in type II cells. PMID- 12629020 TI - Risk-benefit value of inhaled corticosteroids: a pharmacokinetic/pharmacodynamic perspective. PMID- 12629021 TI - Airway response to low-dose allergen exposure in allergic nonasthmatic and asthmatic subjects: eosinophils, fibronectin, and vascular endothelial growth factor. PMID- 12629022 TI - Immunomodulatory effect of interleukin-16 on allergic airway inflammation. PMID- 12629023 TI - Learning occurs with repetitions of inspiratory loading. PMID- 12629024 TI - Regulation of phospholipase A2 by interleulin-1 in human airway smooth muscle. PMID- 12629025 TI - Identification of birch pollen respirable particles. PMID- 12629026 TI - The effect of house dust mite aeroallergen and air pollutant exposures during infancy. PMID- 12629027 TI - Pediatric asthma: an approach to pharmacogenetics analysis. PMID- 12629028 TI - Polymorphisms of the GM-CSF genes and the development of atopic diseases in at risk children. PMID- 12629029 TI - New and exploratory therapies for asthma. PMID- 12629030 TI - Interleukin-13 and interferon modulators of dendritic cell migration in the lungs. PMID- 12629031 TI - Inhibition of airway inflammation and hyperreactivity by a catalytic antioxidant. PMID- 12629032 TI - Zinc status modulates bronchopulmonary eosinophil infiltration in a murine model of allergic inflammation. PMID- 12629033 TI - Asthma and the human genome project: summary of the 45th Annual Thomas L. Petty Aspen Lung Conference. PMID- 12629034 TI - History of hypertension and enhanced thrombogenic activity in postinfarction patients. AB - Hypertension is a risk factor for coronary thrombosis and death in cardiac patients mediated in part by endothelial damage or dysfunction and increased thrombogenicity. However, there are no data regarding the association between hypertension and thrombogenic activity in stable patients after myocardial infarction and limited data about the prognostic significance of thrombogenic factors in hypertensive patients after infarction. Therefore, levels of thrombogenic, lipid, and inflammatory factors were measured 2 months after an acute myocardial infarction in 461 hypertensive and 582 nonhypertensive patients. Thrombogenic factors included d-dimer, fibrinogen, plasminogen activator inhibitor-1, von Willebrand factor, factor VII, and factor VIIa. Lipid variables included cholesterol (total, HDL, LDL), triglyceride, lipoprotein (a), apolipoprotein-A1, and apolipoprotein-B. The prognostic significance of these factors for predicting cardiac events during a 2-year follow-up was evaluated in hypertensive and nonhypertensive patients. In comparison with nonhypertensive patients, those with hypertension had higher levels of d-dimer (607 versus 453 mg/L, P<0.001), fibrinogen (3.64 versus 3.43 g/L, P<0.001), plasminogen activator inhibitor-1 (29.7 versus 27.3 ng/mL, P=0.01), von Willebrand factor (159 versus 141 IU/dL; P<0.001), and higher levels of inflammatory markers (hsCRP and SAA). In multivariate analysis after adjustment for clinical covariates, elevated d dimer was the only factor independently associated with a history of hypertension (OR, 1.38, P=0.05). d-Dimer was associated with an increased risk of recurrent cardiac events in both hypertensive (hazard ratio=3.02, P=0.005) and nonhypertensive (hazard ratio=2.42, P=0.02) patients. Thus, patients after infarction with a history of hypertension have enhanced thrombogenic activity, which predisposes them to recurrent cardiac events. PMID- 12629035 TI - PI3-kinase-induced hyperreactivity in DOCA-salt hypertension is independent of GSK-3 activity. AB - Phosphatidylinositol 3-kinase (PI3K) activity is increased in aortae from deoxycorticosterone (DOCA)-salt rats and enhanced PI3K activity contributes to the arterial hyperreactivity in these animals. Because PI3K activity is increased in DOCA-salt hypertension, we postulated that phosphorylation of Akt and glycogen synthase kinase 3 (GSK-3), serine threonine kinases that are downstream of PI3K, would be increased in DOCA-salt hypertension. In this study, we focused on GSK-3. Because GSK-3 activity is reduced by phosphorylation, we expected that its activity would be reduced in DOCA-salt hypertensive arteries and that reduced GSK 3 activity could contribute to enhanced adrenergic signaling and vascular smooth muscle hypertrophy that augment the heightened contractile response in DOCA-salt hypertension. Surprisingly, we observed a decrease in phosphorylation of GSK-3, indicating an increase in GSK-3 activity. To determine whether increased GSK-3 activity contributes to altered arterial reactivity in DOCA-salt animals, we measured isometric contraction to norepinephrine (NE) in the presence and absence of PI3K or GSK-3 inhibition. Addition of LY294002 (20 micromol/L), a PI3K inhibitor, resulted in a rightward shift in response to NE and normalized the NE induced contractions in the DOCA hypertensive vessels. SB415286, a GSK-3 inhibitor, resulted in a slight rightward shift in response to NE in the DOCA salt vessels. Thus, enhanced GSK-3 activity modestly augments the effects of PI3K but does not appear to contribute greatly to the altered arterial reactivity in DOCA-salt hypertension. PMID- 12629036 TI - NADPH oxidase activity in preeclampsia with immortalized lymphoblasts used as models. AB - Upon activation, neutrophils release reactive oxygen species that are believed to contribute to the widespread manifestation of preeclampsia. Neutrophils have an NADPH oxidase enzyme that catalyzes the production of reactive oxygen species. Little is known about the manifestations of the activated response and the upstream signaling pathways that regulate this process in preeclampsia. It is hypothesized that genetic factors may contribute to the release of reactive oxygen species and consequently the pathophysiology of the disease. We used Epstein-Barr virus-immortalized lymphoblasts from third-trimester, preeclamptic, postpartum preeclamptic women and their respective control subjects to assess NADPH oxidase-mediated reactive oxygen species production by using luminol derived chemiluminescence and dihydrorhodamine-123 fluorescence. There was no effect of pregnancy status on the lymphoblast phorbol ester-stimulated luminol chemiluminescence area under the curve. However, lymphoblasts from preeclamptic patients had significant elevation of the lymphoblast phorbol ester-stimulated luminol area under the curve (F statistic 10.922, P<0.002). Similar findings were evident with dihydrorhodamine-123. No differences were revealed between preeclamptic and control cells when measuring the abundance of the phox proteins using Western blotting. Studies with genistein and tyrphostin implicated tyrosine kinase-dependent mechanisms in the control of NADPH oxidase-associated increased reactive oxygen species production in preeclampsia. These data show that preeclampsia is associated with a predisposition to increased agonist-stimulated NADPH oxidase-mediated reactive oxygen species production. The enhancement of reactive oxygen species generation may be important in mediating the endothelial dysfunction seen in preeclampsia. PMID- 12629037 TI - Long-acting Ca2+ blockers prevent myocardial remodeling induced by chronic NO inhibition in rats. AB - Chronic inhibition of nitric oxide (NO) synthesis induces cardiac remodeling independent of systemic hemodynamic changes in rats. We examined whether long acting dihydropyridine calcium channel blockers block myocardial remodeling and whether the activation of 70-kDa S6 kinase (p70S6K) and extracellular signal regulated kinase (ERK) are involved. Ten groups of Wistar-Kyoto rats underwent 8 weeks of drug treatment consisting of a combination of NO synthase inhibitor NG nitro-l-arginine methyl ester (L-NAME), an inactive isomer (D-NAME), amlodipine (1 or 3 mg/kg per day), or benidipine (3 or 10 mg/kg per day). In other groups, L NAME was also used in combination with a p70S6K inhibitor (rapamycin), a MEK inhibitor (PD98059), and hydralazine. Systolic blood pressure (SBP), heart rate, and left ventricular weight (LVW) were measured, together with histological examinations and kinase assay. L-NAME increased SBP and LVW (1048+/-22 versus 780+/-18 mg, P<0.01) compared with the control, showing a significant increase in cross-sectional area of cardiomyocytes after 8 weeks. Amlodipine, benidipine, or hydralazine equally attenuated the increase in SBP induced by L-NAME. However, both amlodipine and benidipine but not hydralazine attenuated the increase in LVW by L-NAME (789+/-27, 825+/-20 mg, P<0.01, and 1118+/-29 mg, NS, respectively), also confirmed by histological analysis. L-NAME caused a 2.2-fold/1.8-fold increase in p70S6K/ERK activity in myocardium compared with the control, both of which were attenuated by both amlodipine and benidipine but not hydralazine. Both rapamycin and PD98059 attenuated cardiac hypertrophy in this model. Thus, long acting dihydropyridine calcium channel blockers inhibited cardiac hypertrophy induced by chronic inhibition of NO synthesis by inhibiting both p70S6K and ERK in vivo. PMID- 12629038 TI - Molecular imaging in living subjects: seeing fundamental biological processes in a new light. PMID- 12629039 TI - The RAG1 N-terminal domain is an E3 ubiquitin ligase. AB - RAG1 and RAG2 initiate V(D)J recombination, which is the assembly of immunoglobulin and T cell receptor genes. The N-terminal region of RAG1 can be deleted, leaving an enzymatic "core" able to catalyze the complete reaction. Here we report that the N-terminal portion of RAG1 has a distinct enzymatic role separate from the rest of the protein. It acts as an E3 ligase in the ubiquitylation of a test substrate and formation of polyubiquitin chains in vitro. This finding suggests a new way in which V(D)J recombination can be regulated and coupled to other aspects of cell physiology. PMID- 12629040 TI - The nucleotides responsible for the direct physical contact between the chromatin insulator protein CTCF and the H19 imprinting control region manifest parent of origin-specific long-distance insulation and methylation-free domains. AB - The repression of the maternally inherited Igf2 allele has been proposed to depend on a methylation-sensitive chromatin insulator organized by the 11 zinc finger protein CTCF at the H19 imprinting control region (ICR). Here we document that point mutations of the nucleotides in physical contact with CTCF within the endogenous H19 ICR lead to loss of CTCF binding and Igf2 imprinting only when passaged through the female germline. This effect is accompanied by a significant loss of methylation protection of the maternally derived H19 ICR. Because CTCF interacts with other imprinting control regions, it emerges as a central factor responsible for interpreting and propagating gamete-derived epigenetic marks and for organizing epigenetically controlled expression domains. PMID- 12629041 TI - Enhancer-promoter communication mediated by Chip during Pannier-driven proneural patterning is regulated by Osa. AB - The GATA factor Pannier activates proneural achaete/scute (ac/sc) expression during development of the sensory organs of Drosophila through enhancer binding. Chip bridges Pannier with the (Ac/Sc)-Daughterless heterodimers bound to the promoter and facilitates the enhancer-promoter communication required for proneural development. We show here that this communication is regulated by Osa, which is recruited by Pannier and Chip. Osa belongs to Brahma chromatin remodeling complexes and we show that Osa negatively regulates ac/sc. Consequently, Pannier and Chip also play an essential role during repression of proneural gene expression. Our study suggests that altering chromatin structure is essential for regulation of enhancer-promoter communication. PMID- 12629042 TI - Retraction of the Drosophila germ band requires cell-matrix interaction. AB - Integrins and laminins are important mediators of cell-matrix interactions in both vertebrates and invertebrates. Here, we show that germ-band retraction in the Drosophila embryo, during which the tail end of the embryo retracts to its final posterior position, allows the investigation of cell spreading and lamellipodia formation in real time in vivo. We demonstrate that alpha1, 2 laminin and alphaPS3betaPS integrin are required for the spreading of a small group of cells of the amnioserosa epithelium over the tail end of the germ band. We further implicate a role for this spreading in the process of germ-band retraction. PMID- 12629043 TI - Inactivation of Exonuclease 1 in mice results in DNA mismatch repair defects, increased cancer susceptibility, and male and female sterility. AB - Exonuclease 1 (Exo1) is a 5'-3' exonuclease that interacts with MutS and MutL homologs and has been implicated in the excision step of DNA mismatch repair. To investigate the role of Exo1 in mammalian mismatch repair and assess its importance for tumorigenesis and meiosis, we generated an Exo1 mutant mouse line. Analysis of Exo1(-/-) cells for mismatch repair activity in vitro showed that Exo1 is required for the repair of base:base and single-base insertion/deletion mismatches in both 5' and 3' nick-directed repair. The repair defect in Exo1(-/-) cells also caused elevated microsatellite instability at a mononucleotide repeat marker and a significant increase in mutation rate at the Hprt locus. Exo1(-/-) animals displayed reduced survival and increased susceptibility to the development of lymphomas. In addition, Exo1(-/-) male and female mice were sterile because of a meiotic defect. Meiosis in Exo1(-/-) animals proceeded through prophase I; however, the chromosomes exhibited dynamic loss of chiasmata during metaphase I, resulting in meiotic failure and apoptosis. Our results show that mammalian Exo1 functions in mutation avoidance and is essential for male and female meiosis. PMID- 12629044 TI - Essential and dispensable roles of ATR in cell cycle arrest and genome maintenance. AB - A Cre/lox-conditional mouse line was generated to evaluate the role of ATR in checkpoint responses to ionizing radiation (IR) and stalled DNA replication. We demonstrate that after IR treatment, ATR and ATM each contribute to early delay in M-phase entry but that ATR regulates a majority of the late phase (2-9 h post IR). Double deletion of ATR and ATM eliminates nearly all IR-induced delay, indicating that ATR and ATM cooperate in the IR-induced G2/M-phase checkpoint. In contrast to the IR-induced checkpoint, checkpoint delay in response to stalled DNA replication is intact in ATR knockout cells and ATR/ATM and ATR/p53 double knockout cells. The DNA replication checkpoint remains intact in ATR knockout cells even though the checkpoint-stimulated inhibitory phosphorylation of Cdc2 on T14/Y15 and activating phosphorylation of the Chk1 kinase no longer occur. Thus, incomplete DNA replication in mammalian cells can prevent M-phase entry independently of ATR and inhibitory phosphorylation of Cdc2. When DNA replication inhibitors are removed, ATR knockout cells proceed to mitosis but do so with chromosome breaks, indicating that ATR provides a key genome maintenance function in S phase. PMID- 12629045 TI - Suppression of Ras-stimulated transformation by the JNK signal transduction pathway. AB - The c-Jun NH(2)-terminal kinase (JNK) phosphorylates and activates members of the activator protein-1 (AP-1) group of transcription factors and is implicated in oncogenic transformation. To examine the role of JNK, we investigated the effect of JNK deficiency on Ras-stimulated transformation. We demonstrate that although JNK does play a role in transformation in vitro, JNK is not required for tumor development in vivo. Importantly, the loss of JNK expression resulted in substantial increases in the number and growth of tumor nodules in vivo. Complementation assays demonstrated that this phenotype was caused by JNK deficiency. These data demonstrate that, in contrast to expectations, the normal function of JNK may be to suppress tumor development in vivo. This conclusion is consistent with the presence in human tumors of loss-of-function mutations in the JNK pathway. PMID- 12629046 TI - Facilitation of dendritic mRNA transport by CPEB. AB - In neurons, the proteins derived from mRNAs localized in dendrites have been implicated in synaptic plasticity. The cytoplasmic polyadenylation element (CPE), a cis element in the 3'-UTRs of specific dendritic mRNAs, promotes cytoplasmic polyadenylation-induced translation in response to synaptic stimulation. Here, we demonstrate that the CPE and its binding protein CPEB facilitate mRNA transport to dendrites. In rat hippocampal neurons infected with recombinant viruses, the CPE is sufficient to direct a reporter RNA into dendrites. CPEB-GFP protein forms RNA-containing particles that are transported into dendrites in a microtubule dependent fashion at an average velocity of 4-8 microm/min. Such particles also contain maskin, a CPEB-associated factor that mediates cap-dependent translational repression of CPE-containing mRNA, and the molecular motors dynein and kinesin. Overexpression of CPEB in neurons promotes the transport of CPE containing endogenous MAP2 mRNA to dendrites, whereas overexpression of a mutant CPEB that is defective for interaction with molecular motors inhibits this transport. In neurons derived from CPEB knockout mice, the dendritic transport of a CPE-containing reporter RNA is reduced. These results suggest a mechanism whereby CPE-containing mRNAs can be transported to dendrites in a translationally dormant form, but activated at synapses in response to NMDA receptor stimulation. PMID- 12629047 TI - Phosphorylation of RNA polymerase II CTD regulates H3 methylation in yeast. AB - Histone methylation is now realized to be a pivotal regulator of gene transcription. Although recent studies have shed light on a trans-histone regulatory pathway that controls H3 Lys 4 and H3 Lys 79 methylation in Saccharomyces cerevisiae, the regulatory pathway that affects Set2-mediated H3 Lys 36 methylation is unknown. To determine the functions of Set2, and identify factors that regulate its site of methylation, we genomically tagged Set2 and identified its associated proteins. Here, we show that Set2 is associated with Rbp1 and Rbp2, the two largest subunits of RNA polymerase II (RNA pol II). Moreover, we find that this association is specific for the interaction of Set2 with the hyperphosphorylated form of RNA pol II. We further show that deletion of the RNA pol II C-terminal domain (CTD) kinase Ctk1, or partial deletion of the CTD, results in a selective abolishment of H3 Lys 36 methylation, implying a pathway of Set2 recruitment to chromatin and a role for H3 Lys 36 methylation in transcription elongation. In support, chromatin immunoprecipitation assays demonstrate the presence of Set2 methylation in the coding regions, as well as promoters, of genes regulated by Ctk1 or Set2. These data document a new link between histone methylation and the transcription apparatus and uncover a regulatory pathway that is selective for H3 Lys 36 methylation. PMID- 12629048 TI - Protein:protein interactions and the pairing of boundary elements in vivo. AB - Although it is now well-established that boundary elements/insulators function to subdivide eukaryotic chromosomes into autonomous regulatory domains, the underlying mechanisms remain elusive. One idea is that boundaries act as barriers, preventing the processive spreading of "active" or "silenced" chromatin between domains. Another is that the partitioning into autonomous functional units is a consequence of an underlying structural subdivision of the chromosome into higher order "looped" domains. In this view, boundaries are thought to delimit structural domains by interacting with each other or with some other nuclear structure. The studies reported here provide support for the looped domain model. We show that the Drosophila scs and scs' boundary proteins, Zw5 and BEAF, respectively, interact with each other in vitro and in vivo. Moreover, consistent with idea that this protein:protein interaction might facilitate pairing of boundary elements, we find that that scs and scs' are in close proximity to each other in Drosophila nuclei. PMID- 12629049 TI - Association of diacylglycerol kinase zeta with protein kinase C alpha: spatial regulation of diacylglycerol signaling. AB - Activation of PKC depends on the availability of DAG, a signaling lipid that is tightly and dynamically regulated. DAG kinase (DGK) terminates DAG signaling by converting it to phosphatidic acid. Here, we demonstrate that DGKzeta inhibits PKCalpha activity and that DGK activity is required for this inhibition. We also show that DGKzeta directly interacts with PKCalpha in a signaling complex and that the binding site in DGKzeta is located within the catalytic domain. Because PKCalpha can phosphorylate the myristoylated alanine-rich C-kinase substrate (MARCKS) motif of DGKzeta, we tested whether this modification could affect their interaction. Phosphorylation of this motif significantly attenuated coimmunoprecipitation of DGKzeta and PKCalpha and abolished their colocalization in cells, indicating that it negatively regulates binding. Expression of a phosphorylation-mimicking DGKzeta mutant that was unable to bind PKCalpha did not inhibit PKCalpha activity. Together, our results suggest that DGKzeta spatially regulates PKCalpha activity by attenuating local accumulation of signaling DAG. This regulation is impaired by PKCalpha-mediated DGKzeta phosphorylation. PMID- 12629050 TI - AMIGO, a transmembrane protein implicated in axon tract development, defines a novel protein family with leucine-rich repeats. AB - Ordered differential display identified a novel sequence induced in neurons by the neurite-promoting protein amphoterin. We named this gene amphoterin-induced gene and ORF (AMIGO), and also cloned two other novel genes homologous to AMIGO (AMIGO2 and AMIGO3). Together, these three AMIGOs form a novel family of genes coding for type I transmembrane proteins which contain a signal sequence for secretion and a transmembrane domain. The deduced extracellular parts of the AMIGOs contain six leucine-rich repeats (LRRs) flanked by cysteine-rich LRR NH2- and COOH-terminal domains and by one immunoglobulin domain close to the transmembrane region. A substrate-bound form of the recombinant AMIGO ectodomain promoted prominent neurite extension in hippocampal neurons, and in solution, the same AMIGO ectodomain inhibited fasciculation of neurites. A homophilic and heterophilic binding mechanism is shown between the members of the AMIGO family. Our results suggest that the members of the AMIGO protein family are novel cell adhesion molecules among which AMIGO is specifically expressed on fiber tracts of neuronal tissues and participates in their formation. PMID- 12629051 TI - Granzyme B activates procaspase-3 which signals a mitochondrial amplification loop for maximal apoptosis. AB - Granzyme B (GrB), acting similar to an apical caspase, efficiently activates a proteolytic cascade after intracellular delivery by perforin. Studies here were designed to learn whether the physiologic effector, GrB-serglycin, initiates apoptosis primarily through caspase-3 or through BH3-only proteins with subsequent mitochondrial permeabilization and apoptosis. Using four separate cell lines that were either genetically lacking the zymogen or rendered deficient in active caspase-3, we measured apoptotic indices within whole cells (active caspase-3, mitochondrial depolarization [DeltaPsim] and TUNEL). Adhering to these conditions, the following were observed in targets after GrB delivery: (a) procaspase-3-deficient cells fail to display a reduced DeltaPsim and DNA fragmentation; (b) Bax/Bak is required for optimal DeltaPsim reduction, caspase-3 activation, and DNA fragmentation, whereas BID cleavage is undetected by immunoblot; (c) Bcl-2 inhibits GrB-mediated apoptosis (reduced DeltaPsim and TUNEL reactivity) by blocking oligomerization of caspase-3; and (d) in procaspase 3-deficient cells a mitochondrial-independent pathway was identified which involved procaspase-7 activation, PARP cleavage, and nuclear condensation. The data therefore support the existence of a fully implemented apoptotic pathway initiated by GrB, propagated by caspase-3, and perpetuated by a mitochondrial amplification loop but also emphasize the presence of an ancillary caspase dependent, mitochondria-independent pathway. PMID- 12629052 TI - PKA phosphorylation activates the calcium release channel (ryanodine receptor) in skeletal muscle: defective regulation in heart failure. AB - The type 1 ryanodine receptor (RyR1) on the sarcoplasmic reticulum (SR) is the major calcium (Ca2+) release channel required for skeletal muscle excitation contraction (EC) coupling. RyR1 function is modulated by proteins that bind to its large cytoplasmic scaffold domain, including the FK506 binding protein (FKBP12) and PKA. PKA is activated during sympathetic nervous system (SNS) stimulation. We show that PKA phosphorylation of RyR1 at Ser2843 activates the channel by releasing FKBP12. When FKB12 is bound to RyR1, it inhibits the channel by stabilizing its closed state. RyR1 in skeletal muscle from animals with heart failure (HF), a chronic hyperadrenergic state, were PKA hyperphosphorylated, depleted of FKBP12, and exhibited increased activity, suggesting that the channels are "leaky." RyR1 PKA hyperphosphorylation correlated with impaired SR Ca2+ release and early fatigue in HF skeletal muscle. These findings identify a novel mechanism that regulates RyR1 function via PKA phosphorylation in response to SNS stimulation. PKA hyperphosphorylation of RyR1 may contribute to impaired skeletal muscle function in HF, suggesting that a generalized EC coupling myopathy may play a role in HF. PMID- 12629053 TI - Skeletal muscle repair by adult human mesenchymal stem cells from synovial membrane. AB - We have demonstrated previously that adult human synovial membrane-derived mesenchymal stem cells (hSM-MSCs) have myogenic potential in vitro (De Bari, C., F. Dell'Accio, P. Tylzanowski, and F.P. Luyten. 2001. Arthritis Rheum. 44:1928 1942). In the present study, we have characterized their myogenic differentiation in a nude mouse model of skeletal muscle regeneration and provide proof of principle of their potential use for muscle repair in the mdx mouse model of Duchenne muscular dystrophy. When implanted into regenerating nude mouse muscle, hSM-MSCs contributed to myofibers and to long term persisting functional satellite cells. No nuclear fusion hybrids were observed between donor human cells and host mouse muscle cells. Myogenic differentiation proceeded through a molecular cascade resembling embryonic muscle development. Differentiation was sensitive to environmental cues, since hSM-MSCs injected into the bloodstream engrafted in several tissues, but acquired the muscle phenotype only within skeletal muscle. When administered into dystrophic muscles of immunosuppressed mdx mice, hSM-MSCs restored sarcolemmal expression of dystrophin, reduced central nucleation, and rescued the expression of mouse mechano growth factor. PMID- 12629054 TI - Prophylactic treatment of mycotic mucositis in radiotherapy of patients with head and neck cancers. AB - BACKGROUND: Patients undergoing radiotherapy for head and neck cancer are at increased risk of developing oral candidiasis. The objective of this study was to investigate the clinical Candida mucositis and interruptions in radiotherapy in patients suffering from head and neck cancer, receiving fluconazole in comparison with a control group without specific prophylaxis. METHODS: Eighty consecutive patients were randomized in a prospective double-blind trial of prophylactic oral fluconazole or treatment with the same drug when mycotic infections appeared. Adult head and neck cancer patients who were undergoing treatment with radiotherapy and/or chemotherapy, radiotherapeutic coverage of the entire oropharynx and oral cavity at least 3 cm anterior to the retromolar trigone and receiving a total dose of more than 6000 cGy and Karnofsky Performance Status (KPS) >70 were included in the study. Group A received radiation therapy plus fluconazole (Fluzole 100 mg/day) starting from the sixth irradiation session throughout the treatment; 40 patients in group B received the same baseline treatment, but were given fluconazole only when mycotic infections appeared. RESULTS: We evaluated 37 patients in group A and the first 37 patients were evaluated in group B. Three of the patients in group A (8.1%) and 14 of the patients in group B (37.8%) demonstrated clinical candidasis. Radiotherapy was interrupted in all of these patients. The differences between the two groups were statistically significant with respect to clinical candidiasis (P = 0.005). The median discontinuation time was 5 days (range, 3-7 days) in group A and 7 days (range, 4-10 days) in group B. The median dose resulting in clinical candidiasis was 3200 cGy (range, 2200-5800 cGy) in all groups. In the fluconazole group it was 4200 cGy and in the control group 2800 cGy. CONCLUSION: These results suggest that patients undergoing head and neck radiation therapy are at risk of developing candidiasis and that fluconazole may be used to reduce the frequency of mycotic infections and to improve the radiotherapy schedule in head and neck cancer patients. PMID- 12629055 TI - Long-term results of breast-conserving treatment for early-stage breast cancer in Japanese women from multicenter investigation. AB - BACKGROUND: Although many clinical data regarding breast-conserving treatment have already been reported from European and North American countries, few clinical data with long-term follow-up have been reported from Japan. METHOD: We collected information on therapeutic and possible or developed prognostic factors and follow-up data for Japanese women who had received breast-conserving treatment consisting of wide excision of the primary tumor, axillary dissection and radiotherapy for unilateral breast cancer considered suitable for breast conserving treatment from 18 Japanese major breast cancer treating hospitals; 1561 patients were registered. RESULTS: The median follow-up period was 77 months. Five-year disease-free and overall survival rates were 89.4 and 95.9%, respectively. The 5-year local recurrence-free rate was 96.3%. The patients with histologically positive margins (P < 0.0001) or estrogen receptor negative tumor (P = 0.0340) or younger than 40 years old (P < 0.0001) developed statistically significantly more local recurrences. Adjuvant endocrine therapy was essential for the estrogen receptor positive patients to have a lower local recurrence rate. Endocrine therapy did not change the local recurrence rate among estrogen receptor negative patients at all. Multivariate analysis showed histological margin status and the combination of estrogen receptor status and endocrine therapy were independent prognostic factors for local recurrence. CONCLUSION: The 5-year local recurrence rate of Japanese breast cancer patients who were treated with breast-conserving treatment using radiotherapy was 3.7%. Independent prognostic factors for local recurrence were histological margin status and the combination of estrogen receptor status and adjuvant endocrine therapy. PMID- 12629056 TI - Sentinel lymph node detection with Tc-99m tin colloids in patients with esophagogastric cancer. AB - BACKGROUND: The aim of this study was to determine by radioisotope use whether the sentinel lymph node concept is applicable to esophagogastric cancers. In addition, we examined radioactivities of hot nodes and compared them with the sensitivity of a gamma probe. METHODS: The subjects were 44 patients, 23 with esophageal cancer and 21 with gastric cancer. The day before surgery, patients underwent endoscopic submucosal injection of 184 MBq of Tc-99m tin colloids into sites surrounding the tumor. Radioisotope activities of lymph nodes dissected at surgery were measured with a well-typed gamma detector and each lymph node was categorized as a hot or cold node. Histopathology of the lymph nodes was examined by hematoxylin and eosin staining. Radioisotope activities and histopathological results were compared to determine whether radioisotope flow reflects lymphatic flow to regional lymph nodes. The sensitivity of a gamma probe was measured in a laboratory study and the relation between the radioisotope activities of hot nodes and the detection sensitivity of the gamma probe was examined. RESULTS: Histopathological examination revealed lymph node metastasis in 18 of the 44 patients. In 15 of these 18 patients, metastatic foci were recognized in at least one hot node. Subsequent analysis was performed on the 36 patients in whom tumor invasion was confined to the muscle layer and in whom endoscopic clippings had not been applied. Lymph node metastases were observed in 12 of these 36 patients. In these 12 patients, at least one hot node was positive for metastasis. The laboratory study revealed that the gamma probe was able to detect radioisotope activities of >/=0.02 micro Ci. Thirty-two of 63 (51%) esophageal cancer hot nodes and 16 of 86 (19%) gastric cancer hot nodes showed radioisotope activities below the detection sensitivity of the gamma probe. CONCLUSION: The sentinel lymph node concept is applicable to patients with esophageal and gastric cancers; however, further studies are necessary to identify hot nodes accurately using gamma probes. PMID- 12629057 TI - External beam radiation monotherapy for localized or locally advanced prostate cancer. AB - PURPOSE: We report the treatment results and complications of external beam radiation monotherapy for localized or locally advanced prostate cancer patients. METHODS: Fifty-four patients with T(1b-3a)N(0)(pN(0))M(0) prostate cancer were treated with external beam radiation monotherapy between 1989 and 2001 at four institutes. RESULTS: During the 4-122 month follow-up period (median: 25 months), 11 (20%) patients experienced biochemical failure, including one with simultaneous local recurrence. The 2-year actuarial biochemical control rate was 85%. Univariate analysis showed that the clinical T classification (P = 0.01), Gleason score (P = 0.006), pretreatment PSA (P = 0.02) and PSA nadir value (P = 0.01) were associated with a higher probability of biochemical failure. Multivariate analysis using the Cox proportional hazards model demonstrated that only the PSA nadir value was a strong predictor of PSA recurrence (P < 0.01). Adverse events were mild and tolerable. No severe urinary or bowel complications were observed. CONCLUSIONS: External beam radiation monotherapy is effective for clinically organ-confined prostate cancer with a low incidence of severe complications in a mean follow-up period of 2 years. PMID- 12629058 TI - A dose-finding study of glycosylated G-CSF (Lenograstim) combined with CHOP therapy for stem cell mobilization in patients with non-Hodgkin's lymphoma. AB - BACKGROUND: Peripheral blood stem cell (PBSC) reinfusion has been widely used for hematopoietic reconstitution after high-dose chemotherapy. However, the optimal dose of granulocyte colony-stimulating factor (G-CSF) for PBSC mobilization in combination with chemotherapy for autograft remains unknown. METHODS: To find the optimal dose of glycosylated G-CSF (lenograstim) for PBSC mobilization in combination with chemotherapy for aggressive non-Hodgkin's lymphoma (NHL), we conducted a dose-finding study on 43 newly diagnosed patients who had unfavorable prognostic factors. They received four to six courses of cyclophosphamide, doxorubicin, vincristine and prednisolone combined with lenograstim every 2 weeks (biweekly CHOP therapy). PBSC apheresis was started after the third course of biweekly CHOP therapy. Lenograstim was given daily from day 3 until the day of the last apheresis. The optimum dose of lenograstim was assessed based on mobilization efficacy and safety profiles at a daily single dose of 2, 5 and 10 microg/kg for eight patients in each level. RESULTS: The collected number of CD34+ cells in the first apheresis products was higher in the 5 microg/kg group than in the 2 microg/kg group (median, 4.22 x 10(6) vs 2.49 x 10(6) CD34+ cells/kg, P = 0.051). The highest dose of 10 microg/kg (median, 2.99 x 10(6) CD34+ cells/kg) failed to show a dose dependence in PBSC mobilization. The efficacy and safety of the 5 microg/kg dose were further confirmed in an additional 19 patients. CONCLUSIONS: The present study suggests that the recommended dose of lenograstim for PBSC mobilization with CHOP therapy in untreated NHL is 5 microg/kg. PMID- 12629059 TI - Benign metastasizing meningioma. AB - Meningiomas represent about one sixth of all primary neoplasms of the central nervous system. They rarely metastasize outside the intracranial compartment. There are no clear criteria to identify the subset of aggressive tumors which recur locally or metastasize. Histological tumor grade is the most important predictor of recurrence or metastases. We report an unusual case of recurrent metastasis in an extrapleural location and review the literature. Our patient developed recurrent thoracic metastases from an intracranial benign meningioma after a disease-free interval of 8 years. She was successfully managed by wide excision of the metastasis and is currently asymptomatic. PMID- 12629060 TI - A case of subglottic carcinoma effectively treated with intraluminal irradiation using low dose rate iridium-192 thin wires combined with external beam irradiation. AB - Subglottic carcinoma is rare and has a poor prognosis. It is usually treated with surgical resection, external beam irradiation (EBR) and/or a combination of both. Intraluminal irradiation (IR) as a curative treatment for subglottic carcinoma has never been reported. We report a case of subglottic carcinoma in a 66-year old man, who was effectively treated with IR combined with EBR. IR was conducted using low dose rate iridium-192 thin wires at a subglottic mucosal dose of 5.1 6.0 Gy per fraction, with a total dose of 43.8 Gy in eight fractions. EBR was administered, using cobalt-60 and 6 MV X-rays delivered at 2 Gy per fraction, with a total dose of 66 Gy in 33 fractions. Complete response was obtained without serious complications and no recurrence occurred in the 79-month follow up period. IR combined with EBR is useful as a potential curative treatment. It should be considered as one of the treatment options for subglottic carcinoma. PMID- 12629061 TI - Obstructive jaundice in a metastatic tumor of the pancreas from breast cancer: a case report. AB - Metastatic pancreas tumors from breast cancer are comparatively uncommon and patients with this tumor usually remain asymptomatic during their life. A 55-year old woman presented with obstructive jaundice following mastectomy for invasive ductal carcinoma. We diagnosed obstructive jaundice due to a pancreatic tumor demonstrated on computed tomography and performed percutaneous transhepatic cholangio-drainage. Although the patient recovered from the jaundice, she had exacerbation of pneumonia from which she died. At autopsy, invasive ductal carcinoma was found in the pancreas tumor. Immunohistochemical staining was performed to confirm whether the pancreatic tumor was primary or secondary. Human milk fat globules 1 and 2 and gross cystic disease fluid protein-15, which characteristically exist in normal breast tissue or breast carcinoma, were expressed both in the primary breast tumor and the pancreatic tumor. In contrast, both the anti-estrogen receptor and anti-progesterone receptor antibodies stained positively in the primary breast cancer; however, neither of them was positive in the metastatic pancreatic tumor. We report a rare case of a patient who presented with obstructive jaundice from a pancreatic tumor metastasizing from breast cancer and in whom immunohistochemical staining using the antibodies unique to the mammary gland was effective for the diagnosis of this secondary tumor. PMID- 12629062 TI - A comparison of trends in the incidence rate of lung cancer by histological type in the Osaka Cancer Registry, Japan and in the Surveillance, Epidemiology and End Results Program, USA. AB - Trends in the incidence rate of lung cancer by histological type were examined, based on data from the Osaka Cancer Registry (OCR), Japan and the Surveillance, Epidemiology and End Results Program (SEER), USA. Among males, an increasing trend was observed for adenocarcinoma in both registries. In the OCR, the incidence rate of adenocarcinoma has recently overtaken that of squamous cell carcinoma, whereas in the SEER, adenocarcinoma had already taken the lead since 1990-93. For squamous cell carcinoma, the decreasing trends in the OCR were consistent with a previous study, whereas in the SEER, the decline was earlier and steeper. For small cell carcinoma, the incidence rate was leveling off in the OCR, whereas it was already in decline in the SEER. For large cell carcinoma, the decreasing trend began earlier in the SEER. Among females in both registries, the incidence rate of adenocaricinoma has clearly been on the increase, whereas in other histological types the incidence rates were decreasing or at least leveling off. As for the age-specific incidence rate, different patterns were observed between the two registries: in the OCR, the incidence rate was on the increase in older age groups such as 70-74 years and also in younger age groups such as 45-49 years, whereas it decreased in intermediate age groups such as 55-59. In contrast, stable decreasing trends were observed in the SEER, except for adenocarcinoma. The increase in incidence rate observed among younger age groups in the OCR should be carefully monitored. PMID- 12629063 TI - Role of adhesin release for mucosal colonization by a bacterial pathogen. AB - Pathogen attachment is a crucial early step in mucosal infections. This step is mediated by important virulence factors called adhesins. To exert these functions, adhesins are typically surface-exposed, although, surprisingly, some are also released into the extracellular milieu, the relevance of which has previously not been studied. To address the role of adhesin release in pathogenesis, we used Bordetella pertussis as a model, since its major adhesin, filamentous hemagglutinin (FHA), partitions between the bacterial surface and the extracellular milieu. FHA release depends on its maturation by the specific B. pertussis protease SphB1. We constructed SphB1-deficient mutants and found that they were strongly affected in their ability to colonize the mouse respiratory tract, although they adhered even better to host cells in vitro than their wild type parent strain. The defect in colonization could be overcome by prior nasal instillation of purified FHA or by coinfection with FHA-releasing B. pertussis strains, but not with SphB1-producing FHA-deficient strains, ruling out a nonspecific effect of SphB1. These results indicate that the release of FHA is important for colonization, as it may facilitate the dispersal of bacteria from microcolonies and the binding to new sites in the respiratory tract. PMID- 12629064 TI - Biallelic germline transcription at the kappa immunoglobulin locus. AB - Rearrangement of antigen receptor genes generates a vast array of antigen receptors on lymphocytes. The establishment of allelic exclusion in immunoglobulin genes requires differential treatment of the two sequence identical alleles. In the case of the kappa immunoglobulin locus, changes in chromatin structure, methylation, and replication timing of the two alleles are all potentially involved in regulating rearrangement. Additionally, germline transcription of the kappa locus which precedes rearrangement has been proposed to reflect an opening of the chromatin structure rendering it available for rearrangement. As the initial restriction of rearrangement to one allele is critical to the establishment of allelic exclusion, a key question is whether or not germline transcription at the kappa locus is monoallelic or biallelic. We have used a sensitive reverse transcription-polymerase chain reaction (RT-PCR) assay and an RNA-fluorescence in situ hybridization (FISH) to show that germline transcription of the kappa locus is biallelic in wild-type immature B cells and in recombination activating gene (RAG)-/-, mu+ B cells. Therefore, germline transcription is unlikely to dictate which allele will be rearranged first and rather reflects a general opening on both alleles that must be accompanied by a mechanism allowing one of the two alleles to be rearranged first. PMID- 12629065 TI - Treatment of relapsing paralysis in experimental encephalomyelitis by targeting Th1 cells through atorvastatin. AB - Statins, known as inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, exhibit numerous functions related to inflammation, such as MHC class II down-regulation, interference with T cell adhesion, and induction of apoptosis. Here we demonstrate that both subcutaneous and oral administration of atorvastatin inhibit the development of actively induced chronic experimental autoimmune encephalomyelitis in SJL/J mice and significantly reduce the inflammatory infiltration into the central nervous system (CNS). When treatment was started after disease onset, atorvastatin reduced the incidence of relapses and protected from the development of further disability. Both the reduced autoreactive T cell response measured by proliferation toward the encephalitogenic peptide PLP139-151 and the cytokine profile indicate a potent blockade of T helper cell type 1 immune response. In in vitro assays atorvastatin not only inhibited antigen-specific responses, but also decreased T cell proliferation mediated by direct TCR engagement independently of MHC class II and LFA-1. Inhibition of proliferation was not due to apoptosis induction, but linked to a negative regulation on cell cycle progression. However, early T cell activation was unaffected, as reflected by unaltered calcium fluxes. Thus, our results provide evidence for a beneficial role of statins in the treatment of autoimmune attack on the CNS. PMID- 12629066 TI - Fibrin-mediated protection against infection-stimulated immunopathology. AB - Fibrin, a product of the blood coagulation cascade, accompanies many type 1 immune responses, including delayed-type hypersensitivity, autoimmunity, and graft rejection. In those settings, fibrin is thought to exacerbate inflammation and disease. Here, we evaluate roles for coagulation during infection with Toxoplasma gondii, a pathogen whose control requires robust type 1 immunity. We establish that fibrin prevents infection-stimulated blood loss, thereby performing a protective function that is essential for survival. Remarkably, fibrin does not simply protect against vascular damage caused directly by the infectious agent, but rather, protects against hemorrhage evoked by interferon gamma, a critical mediator of type 1 immunity. This finding, to our knowledge, is the first to document a beneficial role for coagulation during type 1 immunity, and suggests that fibrin deposition protects host tissue from collateral damage caused by the immune system as it combats infection. PMID- 12629068 TI - Clinical review 154: The role of pharmacotherapy in perioperative management of patients with acromegaly. PMID- 12629069 TI - Clinical review 155: Pheochromocytoma in Von Hippel-Lindau disease. PMID- 12629070 TI - How should the dose of iodine-131 be determined in the treatment of Graves' hyperthyroidism? PMID- 12629067 TI - Preferential accumulation of antigen-specific effector CD4 T cells at an antigen injection site involves CD62E-dependent migration but not local proliferation. AB - The migration of antigen-specific T cells to nonlymphoid tissues is thought to be important for the elimination of foreign antigens from the body. However, recent results showing the migration of activated T cells into many nonlymphoid tissues raised the possibility that antigen-specific T cells do not migrate preferentially to nonlymphoid tissues containing antigen. We addressed this question by tracking antigen-specific CD4 T cells in the whole body after a localized subcutaneous antigen injection. Antigen-specific CD4 T cells proliferated in the skin-draining lymph nodes and the cells that underwent the most cell divisions acquired the ability to bind to CD62P. As time passed, CD62P binding antigen-specific CD4 T cells with interferon gamma production potential accumulated preferentially at the site of antigen injection but only in recipients that expressed CD62E. Surprisingly, these T cells did not proliferate in the injection site despite showing evidence of more cell divisions than the T cells in the draining lymph nodes. The results suggest that the most divided effector CD4 T cells from the lymph nodes enter the site of antigen deposition via recognition of CD62E on blood vessels and are retained there in a nonproliferative state via recognition of peptide-major histocompatibility complex II molecules. PMID- 12629071 TI - A randomized comparison of radioiodine doses in Graves' hyperthyroidism. AB - The optimal method for determining iodine-131 treatment doses for Graves' hyperthyroidism is unknown, and techniques have varied from a fixed dose to more elaborate calculations based upon gland size, iodine uptake, and iodine turnover. Patients with Graves' hyperthyroidism (n = 88) who had not been previously treated with radioactive iodine were randomized to one of four dose calculation methods: low-fixed, 235 MBq; high-fixed, 350 MBq; low-adjusted, 2.96 MBq (80 micro Ci)/g thyroid adjusted for 24 h radioiodine uptake; and high-adjusted, 4.44 MBq (120 micro Ci)/g thyroid adjusted for 24 h radioiodine uptake. Subjects were followed for mean of 63 months (range, 10-94 months) for the following clinical outcomes: euthyroid without medication, hyperthyroid requiring further radioiodine, and hypothyroid requiring life-long L-T(4). Mean treatment doses were similar in the different outcome groups. We could not demonstrate any advantage to using an adjusted dose method. Survival analysis did not demonstrate any difference in the time to outcome between the fixed and adjusted dose methods. The use of a fixed dose method simplifies the approach to treatment with potential cost savings. PMID- 12629072 TI - Improvement of bone in patients with osteogenesis imperfecta treated with pamidronate-lessons from biochemistry. PMID- 12629073 TI - Osteogenesis imperfecta types I, III, and IV: effect of pamidronate therapy on bone and mineral metabolism. AB - Cyclical iv therapy with pamidronate improves the clinical course in children and adolescents with osteogenesis imperfecta (OI). In this study we evaluated the effect of this therapy on bone and mineral metabolism in 165 patients with OI types I, III, and IV (age, 2 wk to 17.9 yr; 86 girls and 79 boys). All patients received iv pamidronate infusions on 3 successive days, administered at age dependent intervals of 2-4 months. During the 3 d of the first infusion cycle, serum concentrations of ionized calcium dropped by 0.14 +/- 0.008 mmol (mean +/- SE; P < 0.001), and serum PTH levels transiently almost doubled (P < 0.001). At the same time, urinary excretion of the bone resorption marker type I collagen N telopeptide related to creatinine (uNTX/uCr) decreased by 61-73% (P < 0.001). Two to 4 months later, ionized calcium had returned to pretreatment levels, and uNTX/uCr remained 30-35% lower than at baseline (P < 0.001). During 4 yr of pamidronate therapy (n = 40 patients), ionized calcium levels remained stable, but PTH levels increased by about 30% (P < 0.01). uNTX/uCr, expressed as a percentage of the age- and sex-specific mean value in healthy children, decreased from 132 +/- 13% (mean +/- SE) at baseline to 49 +/- 3% after 4 yr of therapy (P < 0.001). In conclusion, serum calcium levels can decrease considerably during and after pamidronate infusions, requiring close monitoring especially at the first infusion cycle. In long-term therapy, bone turnover is suppressed to levels lower than those in healthy children. The consequences of chronically low bone turnover in children with OI are unknown at present. PMID- 12629074 TI - Plasma adiponectin concentrations in relation to endometrial cancer: a case control study in Greece. AB - Adiponectin is a hormone secreted exclusively by adipocytes, and obesity is an established risk factor for endometrial cancer. We have, thus, evaluated the association of adiponectin with the occurrence of endometrial cancer. Questionnaire information and blood samples were taken before treatment from 84 women with newly diagnosed, histologically confirmed endometrial cancer and 84 control women who were admitted for minor gynecologic problems, mainly pelvic prolapse. Adiponectin levels were measured by immunoassay. The results were analyzed through multiple logistic regression and controlled for known risk factors for endometrial cancer, leptin, as well as major components of the IGF system (IGF-I, IGF-II, and IGF-binding protein 3). Among control women, there was no significant association of adiponectin with age or parity. Although there was no association of adiponectin with endometrial cancer among women 65 yr or older, there was an inverse, fairly strong, and statistically significant inverse association among younger women. Among women younger than 65 yr, an increase of adiponectin by 1 SD was associated with a more than 50% reduction of the risk for endometrial cancer [odds ratio (OR) 0.44; 95% confidence interval (CI) 0.24 0.81], even after controlling for body mass index and other potential confounders. Among all women, the adjusted OR for a 1 SD increase in adiponectin was not significant (OR, 0.78; 95% CI, 0.56-1.10) but was significant for a one quintile increase in adiponectin (OR, 0.74; 95% CI, 0.56-0.97). In women younger than 65 yr, among whom obesity represents a powerful risk factor for endometrial cancer, adiponectin is inversely and significantly related to the risk of this disease. This association is independent of possible effects of major components of the IGF system, leptin, body mass index, sociodemographic variables, and known endometrial cancer risk factors. Future studies are needed to prove causality and provide insight on both the mechanism of action of this hormone and its potential role in endometrial cancer. PMID- 12629075 TI - A case report in favor of a multistep adrenocortical tumorigenesis. AB - The mechanisms of adrenocortical tumorigenesis are still not fully understood. Data from clonal analysis, comparative genomic hybridization, and allelotyping suggest that it involves a multistep process during which several genetic defects are progressively acquired, leading to the malignant transformation. The events involved in the first steps of this process are not well known, and most of the abnormalities described in adrenocortical tumors to date are associated with the malignant phenotype. We report a case that suggests that adrenocortical tumorigenesis may be a multistep process. A 43-yr-old patient underwent surgery for an incidentally discovered adrenal mass. Pathological analysis showed that this tumor consisted of two parts: a central part with features of malignancy surrounded by another part with a strictly benign appearance. These data were confirmed by molecular analysis and comparative genomic hybridization that were consistent with either a malignant or benign presentation. The apparently malignant part of the tumor exhibited molecular abnormalities [17p13 loss of heterozygosity (LOH), 11p15 uniparental disomy and overexpression of the IGF-II gene] as well as chromosomal gains and losses (comparative genomic hybridization) that have been previously described in malignant tumors. No abnormalities were found in the surrounding benign tissues. Although this observation is not definitive proof that adrenocortical tumorigenesis occurs via a multistep process, it suggests that there is a progressive change from the benign to the malignant state in some adrenocortical tumors. PMID- 12629076 TI - The W546X mutation of the thyrotropin receptor gene: potential major contributor to thyroid dysfunction in a Caucasian population. AB - Congenital hypothyroidism (CH) occurs in approximately 1 in 3000 births and can be caused by mutations in 9 known genes, including that encoding the TSH receptor (TSHR). We report on two Welsh siblings, detected by neonatal screening, who had normal sized and placed glands but negative isotope uptake. Genomic DNA was obtained from both siblings and parents, the TSHR amplified using pairs of intronic and/or overlapping exonic primers and the PCR products sequenced automatically. Both siblings were homozygous for a previously described G to A transition producing a missense mutation, W546X, in the fourth membrane spanning region of the TSHR, rendering it unresponsive to TSH. Both parents were heterozygous and unrelated; furthermore, the W546X has been described in three further families (one of which is Welsh), suggesting that it may be a relatively common mutation. We genotyped 368 euthyroid Welsh individuals using single nucleotide primer extension, and found 366 homozygous wild-type (G:G) and 2 heterozygous (G:A) for the mutation. In conclusion, CH in the siblings is due to the missense mutation, W546X, in their TSHR gene. The W546X allele was detected in approximately 1 in 180 individuals and may be a major contributor to hypothyroidism in the Welsh population. PMID- 12629077 TI - A new clinical condition linked to a novel mutation in lamins A and C with generalized lipoatrophy, insulin-resistant diabetes, disseminated leukomelanodermic papules, liver steatosis, and cardiomyopathy. AB - A-Type lamins, arising from the LMNA gene, are intermediate filaments proteins that belong to the lamina, a ubiquitous nuclear network. Naturally occurring mutations in these proteins have been shown to be responsible for several distinct diseases that display skeletal and/or cardiac muscle or peripheral nerve involvement. These include familial partial lipodystrophy of the Dunnigan type and the mandibuloacral dysplasia syndrome. The pathophysiology of this group of diseases, often referred to as laminopathies, remains elusive. We report a new condition in a 30-yr-old man exhibiting a previously undescribed heterozygous R133L LMNA mutation. His phenotype associated generalized acquired lipoatrophy with insulin-resistant diabetes, hypertriglyceridemia, hepatic steatosis, hypertrophic cardiomyopathy with valvular involvement, and disseminated whitish papules. Immunofluorescence microscopic analysis of the patient's cultured skin fibroblasts revealed nuclear disorganization and abnormal distribution of A-type lamins, similar to that observed in patients harboring other LMNA mutations. This observation broadens the clinical spectrum of laminopathies, pointing out the clinical variability of lipodystrophy and the unreported possibility of hypertrophic cardiomyopathy and skin involvement. It emphasizes the fact that the diagnosis of genetic alterations in A-type lamins requires careful and complete clinical and morphological investigations in patients regardless of the presenting signs. PMID- 12629078 TI - Age-related changes in cortical bone content of insulin-like growth factor binding protein (IGFBP)-3, IGFBP-5, osteoprotegerin, and calcium in postmenopausal osteoporosis: a cross-sectional study. AB - Serum GH and IGF-I levels decline with increasing age, whereas osteoprotegerin (OPG) increases. IGFs as well as OPG are present in bone matrix and mediate the effects of many upstream hormones (e.g. estrogen). To evaluate whether changes in these proteins may to some extent explain the decrease in bone mass in postmenopausal or senile osteoporosis, we measured bone contents of IGF-I, IGF II, IGF binding protein (IGFBP)-3, IGFBP-5, and OPG in combined extracts obtained after EDTA and guanidine hydrochloride extraction in 60 postmenopausal women aged 47-74 (mean, 63) yr with a previous distal forearm fracture and a hip or spine Z score less than 0. We found age-related increases in IGFBP-3 (r = 0.35; P < 0.01), IGFBP-5 (r = 0.59; P < 0.001), and OPG (r = 0.36; P < 0.01) in cortical bone, significantly inversely correlated with femoral neck and lumbar spine BMD. A correlation between age and OPG was also detected in trabecular bone (r = 0.27; P < 0.05). A pronounced age-related decrease in cortical calcium contents (r = 0.60; P < 0.001), positively correlated with femoral neck and lumbar spine BMD, was also found. No age-related changes were detected for IGF-I or IGF-II. The present study demonstrates age-related changes in cortical bone contents of IGFBPs, calcium, and OPG, possibly related to the pathophysiology of postmenopausal osteoporosis. As for OPG, our findings probably represent compensatory responses to increased osteoclastic resorption. PMID- 12629079 TI - Novel insulin sensitivity index derived from oral glucose tolerance test. AB - The euglycemic hyperglycemic clamp is generally regarded as a reference method for assessing insulin sensitivity. However, this method is laborious and expensive. The oral glucose tolerance test (OGTT), the most commonly used method for evaluating whole body glucose tolerance, has often been used to assess insulin sensitivity. In the previous studies the correlation between the insulin sensitivity index (ISI) obtained from the OGTT (ISI(OGTT)) and those obtained from the glucose clamp (ISI(Clamp)) may not be satisfactory. This is because the glucose clamp study is designed for measuring peripheral glucose utilization, whereas plasma glucose responses during the OGTT are the results of peripheral glucose utilization and hepatic glucose production. Based on this problem, we developed a new equation, ISI(OGTT), [1.9/6 x body weight (kg) x fasting plasma glucose (mmol/liter) + 520 - 1.9/18 x body weight x area under the glucose curve (mmol/h.liter) - urinary glucose (mmol)/1.8] / [area under the insulin curve (pmol/h.liter) x body weight], which would represent peripheral glucose utilization only. We tested our equation with ISI(Clamp) and also compared with others. Thirty-three healthy volunteers (16 males) with normal glucose tolerance underwent a 75-g, 3-h OGTT on the morning of d 1 and a glucose clamp on the morning of d 2. Their mean (+/-SD) age and body mass index were 30.8 +/- 8.3 yr and 22.0 +/- 3.9 kg/m(2), respectively. The mean (+/-SD) glucose disposal rate and ISI determined by glucose clamp were 27.46 +/- 16.55 micro mol/kg.min and 7.39 +/- 2.72 micro mol/kg.min/pmol.liter, respectively. Pearson's correlation coefficient between our ISI(OGTT) and ISI(Clamp) was 0.869 (P < 0.0001) which was stronger than those corresponding values calculated from HOMA, QUICKI, Belfiore, Cederholm, Gutt, Matsuda, and Stumvoll, the respective values of which were 0.404, 0.434, 0.643, 0.533, 0.584, 0.734, and 0.508. In conclusion, the ISI(OGTT) derived from our equation is more suitable than others in assessing insulin sensitivity in subjects with normal glucose tolerance. Further studies in subjects with impaired glucose tolerance and diabetes mellitus should be performed to confirm the validity of this equation. PMID- 12629080 TI - Increased osteoprotegerin serum levels in men with coronary artery disease. AB - Osteoprotegerin (OPG) regulates osteoclast and immune functions and appears to represent a protective factor for the vascular system. However, the role of OPG in human atherosclerosis has not been evaluated. In this study, we assessed OPG serum levels in 522 age-matched men who, on the basis of coronary angiography, had either absence of coronary artery disease (CAD) or presence of single-vessel disease, double-vessel disease, or severe triple-vessel disease. OPG serum levels were positively correlated with age (r = 0.28; P < 0.001) and were higher in men with diabetes mellitus (P < 0.01). OPG serum levels in men without CAD were 5.4 +/- 2.0 pmol/liter, compared with 6.1 +/- 2.1 pmol/liter in single-vessel disease (P < 0.005), 5.9 +/- 2.4 in double-vessel disease (P < 0.05), and 6.3 +/- 2.3 pmol/liter in triple-vessel disease (P < 0.001). Moreover, OPG serum levels were positively correlated with the severity of CAD as determined by a CAD scoring system (r = 0.17; P < 0.01). In conclusion, our data underline that OPG serum levels are associated with the severity of CAD and are increased in elderly men and patients with diabetes mellitus. We conclude that increased OPG serum levels may reflect advanced cardiovascular disease in men. PMID- 12629081 TI - Management of occult adrenocorticotropin-secreting bronchial carcinoids: limits of endocrine testing and imaging techniques. AB - The differential diagnosis and the identification of the source of ACTH in occult ectopic Cushing's syndrome due to a bronchial carcinoid still represents a challenge for the endocrinologist. We report our experience in six patients with occult bronchial carcinoid in whom extensive hormonal, imaging, and scintigraphic evaluation was performed. All patients presented with hypercortisolism associated with high plasma ACTH values. The CRH test and high dose dexamethasone suppression test suggested an ectopic source of ACTH in three of six patients. During bilateral inferior petrosal sinus sampling, none of the patients showed a central to peripheral ACTH gradient. At the time of diagnosis, none of the patients had radiological evidence of the ectopic source of ACTH, whereas pentetreotide scintigraphy identified the lesion in two of four patients. Finally, a chest computed tomography scan revealed the presence of a bronchial lesion in all patients, and pentetreotide scintigraphy identified four of six lesions. In all patients a bronchial carcinoid was found and removed. In one patient with scintigraphic evidence of residual disease after two operations, radioguided surgery, using a hand-held gamma probe after iv administration of radiolabeled pentetreotide, was performed; this allowed detection and removal of residual multiple mediastinal lymph node metastases. In conclusion, our data show that there is not a single endocrine test or imaging procedure accurate enough to diagnose and localize occult ectopic ACTH-secreting bronchial carcinoids. Radioguided surgery appears to be promising in the presence of multiple tumor foci and previous incomplete removal of the tumor. PMID- 12629082 TI - Normal bone mineral density and lean body mass, but increased fat mass, in young adult patients with congenital adrenal hyperplasia. AB - Patients with congenital adrenal hyperplasia attributable to 21-hydroxylase deficiency are treated with glucocorticoids. Glucocorticoid administration, even in substitution doses, may cause decreased bone mineral density (BMD) and obesity. The purpose of this study was to determine BMD, lean mass, and fat mass in young adult male (M, n = 15) and female (F, n = 15) patients with 21 hydroxylase deficiency, who had been treated with currently recommended low doses of glucocorticoids. Measurements were performed with dual-x-ray absorptiometry. In addition, calcaneal ultrasound measurements were performed (broadband ultrasound attenuation and speed of sound). Results were compared with those in age- and sex-matched controls; to adjust for height, lean and fat mass were divided by (height)(2). M and F patients [M, 21.7 +/- 2.4; F, 20.6 +/- 2.9 yr old (mean +/- SD)] were shorter than the controls (M, P < 0.001; F, P < 0.003) and their body mass indices were higher [M patients (25.0 +/- 3.6) vs. controls (22.3 +/- 1.9 kg/m(2)) (P < 0.02); F patients (25.5 +/- 4.5) vs. controls (21.9 +/- 2.3 kg/m(2)) (P < 0.02)]. BMD values (lumbar spine L1-L4, femoral neck, and total body) were not different from controls. Calcaneal ultrasound measurements showed that M patients had higher speed of sound values [M patients (1564 +/- 38) vs. controls (1529 +/- 29 m/sec) (P < 0.01)]. Lean mass in M and F patients was not different from controls when adjusted for height. Fat mass was higher in M and F patients when adjusted for height [M patients 5.6 +/- 2.9 vs. controls 2.7 median (1.7-7.0 min-max) kg/m(2) (P < 0.04); F patients 8.7 +/- 2.8 vs. controls 5.8 (4.3-10.7) kg/m(2) (P < 0.02)]. Relative fat mass (fat mass as a percentage of the total body mass) was higher in patients, compared with controls [M patients 22.0 +/- 9.1 vs. controls 12.8 (8.5-27.0)% (P < 0.04); F patients 34.1 +/- 5.0 vs. controls 29.0 +/- 5.1% (P < 0.02)]; this resulted from increased fat mass, not from decreased lean mass. Fat distribution over the body was not different in patients and controls. No significant correlations were found between cumulative glucocorticoid doses in the last 0.5, 2, or 5 yr or mean salivary morning levels of 17-hydroxyprogesterone and androstenedione in the last 5 yr on one hand and bone parameters, lean mass, or fat mass on the other hand. We conclude that, at prevailing low-dose glucocorticoid regimens, young adult patients with 21 hydroxylase deficiency have normal BMD. Their lean mass is in accordance with height, but fat mass is increased, with a normal distribution over the body. This results in a higher fat percentage of the total body and a higher body mass index than in healthy peers. Because overweight and increased fat mass are associated with the metabolic syndrome and increased cardiovascular risk, weight management should have appropriate attention in the follow-up of congenital adrenal hyperplasia patients, to prevent overweight-associated morbidity. PMID- 12629083 TI - Racial differences in bone turnover and calcium metabolism in adolescent females. AB - Blacks develop a higher peak bone mass than whites which is associated with a reduced risk for bone fracture. The physiological basis for the difference in bone mass was investigated by metabolic balance and calcium kinetic studies in adolescent black and white girls. The hypothesis that the greater peak bone mass in blacks compared with whites is due to suppressed bone resorption was tested. Subjects were housed in a supervised environment for 3 wk during which time they consumed a controlled diet and collected all excreta. Subjects were given stable calcium isotopes orally and intravenously after 1 wk adaptation. Blacks have greater calcium retention (mean +/- SD, 11.5 +/- 6.1 vs. 7.3 +/- 4.1 mmol/d, P < 0.05) consistent with greater bone formation rates (49.4 +/- 13.5 vs. 36.5 +/- 13.6 mmol/d, P < 0.05) relative to bone resorption rates (37.4 +/- 13.2 vs. 29.4 +/- 10.9 mmol/d, P = 0.07), increased calcium absorption efficiency (54 +/- 19 vs. 38 +/- 18%, P < 0.05) and decreased urinary calcium (1.15 +/- 0.95 vs. 2.50 +/- 1.35 mmol/d, P < 0.001), compared with whites. The racial differences in calcium retention in adolescence can account for the racial differences in bone mass of adults. PMID- 12629084 TI - Soy protein has a greater effect on bone in postmenopausal women not on hormone replacement therapy, as evidenced by reducing bone resorption and urinary calcium excretion. AB - Recent reports suggest that soy protein may reduce the risk of osteoporosis in peri- and postmenopausal women. The objective of this study was to examine whether soy supplementation exerts beneficial effects on serum and urinary biomarkers of bone metabolism in postmenopausal women, regardless of whether or not they are on hormone replacement therapy (HRT). A total of 71 women were randomly assigned to either soy protein (SP) or milk-based protein (MBP), 40 g daily for 3 months, in a double-blind parallel design. Forty-two women completed the study (20 on SP and 22 on MBP). Overall, both protein supplements positively influenced serum IGF-I, known to correlate with bone formation. However, SP had a more pronounced effect on IGF-I than MBP. Urinary deoxypyridinoline (Dpd) excretion, a specific biomarker of bone resorption, was significantly reduced by SP, but not by MBP when all women were included. Furthermore, women on MBP experienced a 33% increase in urinary calcium excretion, whereas SP did not have such an effect. To evaluate whether SP affects women differently on the basis of their HRT status, data from women on HRT (n = 22) and those not on HRT (n = 20) were analyzed separately. The subanalysis of the data indicated that SP had the greatest impact on serum IGF-I (an increase of 97%) in the women not on HRT. The changes in urinary Dpd due to SP were only observed in women not on HRT, indicating that the overall decrease in Dpd occurred with SP in the absence of HRT. These results indicate that soy protein may positively influence bone and calcium homeostasis in postmenopausal women, particularly those not on HRT. PMID- 12629085 TI - Association of low interleukin-10 levels with the metabolic syndrome in obese women. AB - The potential role of anti-inflammatory cytokines in human obesity is unknown. We tested the hypothesis that low serum IL-10 concentrations associate with the metabolic syndrome in obese women. Compared with 50 matched nonobese women, the prevalence of the metabolic syndrome (>/=3 of the following abnormalities: waist circumference, >88 cm; triglycerides, >1.69 mmol/liter; high density lipoprotein cholesterol, <1.29 mmol/liter; blood pressure, >130/85 mm Hg; glucose, >6.1 mmol/liter) was higher in 50 obese women (52% vs. 16%; P < 0.01). As a group, obese women had higher circulating levels of IL-6, C-reactive protein, and IL-10 than nonobese women. In both obese and nonobese women, IL-10 levels were lower in those with than in women without the metabolic syndrome: obese, 1.3 (0.7/2.1) pg/ml vs. 4.5 (4.3/7.4) pg/ml (median and quartiles; P < 0.01); and nonobese, 0.9 (0.7/1.3) pg/ml vs. 1.3 (0.9/3.3) pg/ml (P < 0.05). After 12 months of a lifestyle program, body weight decreased by 10.9 +/- 1.7 kg and was associated with a significant decrement of IL-6, C-reactive protein, and IL-10 levels; the decrease in IL-10 levels was confined to obese women without the metabolic syndrome. These results show that circulating levels of the anti-inflammatory cytokine IL-10 are elevated in obese women and that low IL-10 levels are associated with the metabolic syndrome. PMID- 12629086 TI - Serum insulin-like growth factor (IGF)-I and IGF-binding protein-1 in elderly people: relationships with cardiovascular risk factors, body composition, size at birth, and childhood growth. AB - The IGF system is important in regulation of fetal and childhood growth. In later life, IGF-I and IGF-binding protein-1 (IGFBP-1) have been implicated in the pathogenesis of arteriosclerosis. They are, thus, potential candidates in explaining the link between early growth and adult cardiovascular disease. We measured fasting serum IGF-I and IGFBP-1 concentrations in 394 men and women from a cohort of 7086 individuals, born between 1924 and 1933 in Helsinki, Finland, whose weight and height were recorded at birth and from 7 to 15 yr of age. They also underwent clinical examination, including measurement of body fat using bioimpedance, blood pressure, glucose tolerance, and plasma insulin and fibrinogen concentrations. Serum IGF-I was positively correlated with fasting glucose (r = 0.10, P = 0.06) and fibrinogen (r = 0.19, P = 0.0001) concentrations and blood pressure (systolic and diastolic r = 0.10, P 5 d). CRP and MBL concentrations were measured on admission, d 5, d 15, and the last day in the ICU. In all patients, serum MBL concentrations increased with time in the ICU (P < 0.0001). This acute phase response was suppressed by intensive insulin therapy at all time points studied (P < 0.02). Selectively in patients receiving conventional therapy, MBL concentrations at baseline were almost 3 times higher in survivors than in nonsurvivors (P = 0.04). Baseline CRP concentrations were elevated, but decreased with time in ICU (P < 0.0001). The decrease in CRP was significantly more pronounced in the intensive insulin-treated patients compared with the conventionally treated patients (P 15 yr). Total bone mineral content, total BMD, and fat percentage were evaluated at the end of the study period using dual-energy x-ray absorptiometry. Final height was significantly higher than predicted height at diagnosis (159.9 +/- 6.3 cm vs. 152.9 +/- 9.6 cm; P < 0.05). Body mass index and fat percentage were not statistically different from control values. Densitometric values at final evaluation in groups A and B together were lower than in controls, but the differences were not statistically significant. The vBMD was significantly higher in group B than in group A at the end of treatment period (0.213 +/- 0.022 g/cm(3) vs. 0.192 +/- 0.021 g/cm(3); P < 0.01) and at final evaluation (0.246 +/- 0.023 g/cm(3) vs. 0.227 +/- 0.024 g/cm(3); P < 0.05). The percentage change (Delta%) between the start and end of treatment period in AP-BMD and vBMD was significantly higher in group B than in group A (Delta% AP-BMD: 20.36% +/- 1.10% vs. 16.16% +/- 1.90%, P < 0.01; Delta% vBMD: 19.08% +/- 3.52% vs. 9.26% +/- 5.15%; P < 0.01) and also between the start of treatment and final evaluation (Delta% AP-BMD: 61.23% +/- 1.61% vs. 56.97% +/- 1.45%, P < 0.01; Delta% vBMD: 36.69% +/- 5.01% vs. 28.01% +/- 5.76%, P < 0.01). In all our females with CPP treated with GnRHa, bone densitometric parameters were in the normal range for age and sex. However, bone mass achievement seemed to be better preserved in the group of patients supplemented with calcium. PMID- 12629091 TI - Effects on gender identity of prenatal androgens and genital appearance: evidence from girls with congenital adrenal hyperplasia. AB - To address questions about sex assignment in children with ambiguous genitalia, we studied gender identity in girls with congenital adrenal hyperplasia (CAH) in relation to characteristics of the disease and treatment, particularly genital appearance and surgery. A 9-item gender identity interview was administered to 43 girls with classical CAH ranging in age from 3-18 yr, 7 tomboys, and 29 sister control girls. Groups were compared on total score and on individual items. Results showed that, on the total gender identity score, 88% of girls with CAH had scores overlapping those of control girls, but the average score was intermediate between control girls and tomboys. On individual items of gender identity (discomfort as a girl, wish to be a boy), girls with CAH were similar to control girls. Gender identity in girls with CAH was not related to degree of genital virilization or age at which genital reconstructive surgery was done. Thus, moderate androgen excess early in development appears to produce a small increase in the risk of atypical gender identity, but this risk cannot be predicted from genital virilization. PMID- 12629092 TI - Positive predictive value of serum thyroglobulin levels, measured during the first year of follow-up after thyroid hormone withdrawal, in thyroid cancer patients. AB - The follow-up of patients with papillary and follicular thyroid carcinoma after thyroidectomy and radioiodine ablation is mainly based on serum thyroglobulin (Tg) level deter-mination. The positive predictive value (PPV) of serum Tg level after thyroid hormone withdrawal, measured during the first 6-12 months of follow up (initial off L-T(4) Tg), was studied in 256 consecutive differentiated thyroid cancer patients. All underwent a total thyroidectomy and 3.7 GBq (131)I ablation; 37 patients had an elevated initial off L-T(4) Tg level. This study focuses on these 37 patients, 9 of whom had a clinical recurrence. The present data confirm that in this selected cohort of patients, 74-185 MBq (131)I-total body scan (TBS) has no clinical interest in the initial work-up and during the subsequent follow up because it was negative in all patients, except in one with recurrent disease. The PPV of initial serum off L-T(4) Tg level above 5 ng/ml and 10 ng/ml was 42% and 53%, respectively; this PPV was only 50% at the time of recurrence or subsequent control. This relatively low PPV is related to the low recurrence rate in this series of patients, despite a prolonged follow-up, and to the subsequent decrease of serum Tg level in 14 of 37 (38%) patients in the absence of any further treatment. In contrast, the PPV of the increasing slope of serum Tg levels obtained after thyroid hormone withdrawal (83%) was excellent. In conclusion, we confirm that (131)I-TBS has a limited interest for the follow-up of thyroid cancer patients. Follow-up should rely on serum Tg level and prognostic parameters; however, initial serum Tg may be produced by thyroid tissues of various significance, an increase at two consecutive determinations indicating disease progression and a decrease being related to late effects of therapy. The best PPV is brought by the slope of serum Tg levels. PMID- 12629093 TI - Replacement of dehydroepiandrosterone in adrenal failure: no benefit for subjective health status and sexuality in a 9-month, randomized, parallel group clinical trial. AB - The physiological role of dehydroepiandrosterone (DHEA) is not well understood, but studies suggest positive effects on subjective health and bone metabolism. We have conducted a clinical trial with DHEA replacement in adrenal failure with the primary aim of evaluating effects on subjective health status and sexuality. Thirty-nine women with adrenal failure were randomized to 9 months of treatment with 25 mg DHEA (n = 19) or placebo (n = 20). Treatment effects were assessed by validated questionnaires of subjective health and sexuality. DHEA replacement yielded a wide variation of effects on the subjective health scales, which were not different from the effects of placebo. Almost all patients receiving DHEA obtained normal androgen levels. Eighty-nine percent of the patients receiving DHEA experienced side-effects, in particular increased sweat odor and scalp itching. DHEA replacement did not significantly change the levels of blood lipids, IGF-I, and markers of bone metabolism. In conclusion, we do not find evidence of beneficial effects of DHEA on subjective health status and sexuality in adrenal failure. However, DHEA may be beneficial for subgroups of patients with adrenal failure, but these remain to be identified. Premenopausal androgen levels can be restored with 25 mg DHEA daily in most female patients, but side effects are frequent. PMID- 12629095 TI - Association of postpartum thyroid dysfunction with antepartum hormonal and immunological changes. AB - Postpartum thyroid dysfunction (PPTD) develops during the first 9 months in up to 50% of women who have thyroid peroxidase antibodies (anti-TPOAb +ve). Humoral immunity in PPTD has been well documented, but the cellular immunological events accompanying the Th2 to Th1 state postpartum are less clear. Peripheral blood lymphocyte cytokine secretion was examined in 48 TPOAb +ve and 33 TPOAb -ve women at 36 wk gestation and at 6, 12, and 24 wk postpartum. Eighteen women with PPTD had significantly greater secretion of interferon gamma and IL-4 than euthyroid women at 36 wk gestation with no significant differences in cytokine secretion at other time points. Also, at 36 wk gestation, the median plasma cortisol concentration in the PPTD group was significantly lower than the euthyroid group (442 nmol/liter vs. 567 nmol/liter, P < 0.02). There were no differences between the groups in levels of prolactin, progesterone, or estradiol. These data suggest that there may be less immunological suppression at 36 wk in TPOAb +ve women destined to develop PPTD possibly because of lower levels of cortisol. Thus, the immunological determinants of PPTD may in part occur antenatally, although the mechanism(s) for this is still unclear. PMID- 12629094 TI - Final height in girls with turner syndrome after long-term growth hormone treatment in three dosages and low dose estrogens. AB - Although GH treatment for short stature in Turner syndrome is an accepted treatment in many countries, which GH dosage to use and which age to start puberty induction are issues of debate. This study shows final height (FH) in 60 girls with Turner syndrome treated in a randomized dose-response trial, combining GH treatment with low dose estrogens at a relatively young age. Girls were randomly assigned to group A (4 IU/m(2).d; approximately 0.045 mg/kg/d), group B (first year, 4 IU/m(2).d; thereafter 6 IU/m(2).d), or group C (first year, 4 IU/m(2).d; second year, 6 IU/m(2).d; thereafter, 8 IU/m(2).d). After a minimum of 4 yr of GH treatment, at a mean age of 12.7 +/- 0.7 yr, low dose micronized 17beta-estradiol was given orally. After a mean duration of GH treatment of 8.6 +/- 1.9 yr, FH was reached at a mean age of 15.8 +/- 0.9 yr. FH, expressed in centimeters or SD score, was 157.6 +/- 6.5 or -1.6 +/- 1.0 in group A, 162.9 +/- 6.1 or -0.7 +/- 1.0 in group B, and 163.6 +/- 6.0 or -0.6 +/- 1.0 in group C. The difference in FH in centimeters, corrected for height SD score and age at start of treatment, was significant between groups A and B [regression coefficient, 4.1; 95% confidence interval (CI), 1.4, 6.9; P < 0.01], and groups A and C (coefficient, 5.0; 95% CI, 2.3, 7.7; P < 0.001), but not between groups B and C (coefficient, 0.9; 95% CI, -1.8, 3.6). Fifty of the 60 girls (83%) had reached a normal FH (FH SD score, more than -2). After starting estrogen treatment, the decrease in height velocity (HV) changed significantly to a stable HV, without affecting bone maturation (change in bone age/change in chronological age). The following variables contributed significantly to predicting FH SD score: GH dose, height SD score (ref. normal girls), chronological age at start of treatment, and HV in the first year of GH treatment. GH treatment was well tolerated. In conclusion, GH treatment leads to a normalization of FH in most girls, even when puberty is induced at a normal pubertal age. The optimal GH dosage depends on height and age at the start of treatment and first year HV. PMID- 12629096 TI - Modulation of hunger by plasma glucose and metformin. AB - The plasma glucose concentration is a major short-term regulator of hunger and food intake. In patients with diabetes, therapies lowering plasma glucose are frequently associated with body weight gain, suggesting that lowered plasma glucose leads to increased feelings of hunger and food intake. However, as many physiological and symptomatic responses to low plasma glucose are attenuated after repeated episodes of hypoglycemia, this may also pertain to feelings of hunger. Here we tested whether the stimulatory effect of low plasma glucose on feelings of hunger is likewise reduced by repeated episodes of hypoglycemia. As metformin has been shown to reduce plasma glucose levels without increasing body weight and also to decrease food intake, we tested for possible interacting effects of this substance with hypoglycemia-induced hunger. Feelings of hunger were assessed by rating scales during 3 consecutive hypoglycemic clamps performed on 2 consecutive d in 15 normal weight men. Subjects were tested once while being treated with 850 mg metformin twice daily and once while receiving placebo. Treatment was started 14 d before the clamp experiments and was performed in a random order and double-blind fashion. Hypoglycemia markedly enhanced feelings of hunger (P < 0.001). However, rated feelings of hunger on the first and last hypoglycemic clamps were comparable (P = 0.304). Compared with placebo, metformin decreased feelings of hunger during hypoglycemia (P = 0.015). This reduction was not associated with a decrease in posthypoglycemic food intake as measured by the number of cookies consumed after the last clamp (P = 0.676). Data indicate that the stimulatory effect of low plasma glucose on hunger is not attenuated after repeated episodes of hypoglycemia, which implies that, in contrast to other symptoms, hunger is not subject to adaptive attenuation upon repeated hypoglycemia. Metformin attenuates hypoglycemia-induced hunger, but does not appear to influence posthypoglycemic food intake. PMID- 12629097 TI - Effects of recombinant human insulin-like growth factor (IGF)-I and estrogen administration on IGF-I, IGF binding protein (IGFBP)-2, and IGFBP-3 in anorexia nervosa: a randomized-controlled study. AB - Administration of recombinant human (rh) IGF-I has been shown to have positive effects on bone density in anorexia nervosa, but the effects of rhIGF-I and estrogen on IGF binding protein (IGFBP)-2 and IGFBP-3 in anorexia nervosa are not known. Sixty-five osteopenic women with anorexia nervosa were randomized to rhIGF I (30 micro g/kg sc twice daily) alone (n = 15), daily ethinyl estradiol (Ovcon 35) with rhIGF-I (n = 15), estradiol and placebo (n = 15), or placebo (n = 14) for 9 months. Subjects were 25.6 +/- 0.8 yr of age, low weight (body mass index 16.6 +/- 0.2 kg/m(2)) and osteopenic (T scores -2.06 +/- 0.09 for spine and -1.76 +/- 0.13 for hip). IGFBP-3 correlated with total hip bone density (r = 0.47, P = 0.0002) and was a significant predictor of hip bone density (P = 0.010) independent of IGF-I and body mass index in a multivariate regression model. During therapy, IGFBP-2 increased by 48 +/- 19 ng/ml in response to rhIGF-I and decreased by -38 +/- 22 ng/ml in response to placebo (P = 0.011). IGFBP-3 decreased (-895 +/- 120 ng/ml) in response to rhIGF-I but showed a minimal change (-53 +/- 99 ng/ml) in response to placebo (P < 0.0001). In contrast, no significant effect of estrogen was seen on IGF-I, IGFBP-2 or IGFBP-3. Among patients receiving rhIGF-I, the change in IGFBP-2 was inversely associated with the change in total hip bone density (R = -0.47, P = 0.013). In conclusion, our data suggest that chronic rhIGF-I administration increases IGF-I and IGFBP-2 and decreases IGFBP-3 in women with anorexia nervosa. IGFBP-2 and IGFBP-3 may be important determinants of bone density in this population. PMID- 12629098 TI - Effects of intermittent parathyroid hormone administration on bone mineralization density in iliac crest biopsies from patients with osteoporosis: a paired study before and after treatment. AB - Anabolic effects of PTH have been observed at several skeletal sites in humans by dual x-ray absorptiometry without differentiating between an actual increase in bone volume and an increase in mineral content within already established bone. The present study addressed this issue by evaluating the bone mineralization density distribution of iliac crest bone biopsies before and after PTH treatment for 18-36 months in men and women with osteoporosis using quantitative backscattered electron imaging. In cortical bone, pairwise comparison of the two biopsies before and after treatment revealed a reduction in the typical calcium concentration in men (-3.32%; P = 0.02, by paired t test), but no change in women, and the heterogeneity of mineralization increased in both males and females [+18.80% (P = 0.09) and +18.14% (P = 0.005), respectively]. In cancellous bone, there was no change in the typical calcium concentration, but there was a greater heterogeneity of mineralization in both men and women [+19.65% (P = 0.02) and +21.59% (P = 0.056), respectively] due to newly formed bone matrix. Small angle x-ray scattering performed on a subgroup of subjects revealed normal collagen/mineral structure. The findings confirm the observations that PTH stimulates skeletal remodeling, resulting in an increased percentage of newly formed bone matrix of lower mineral density. PMID- 12629099 TI - Relationship between serum leptin concentration and low-density muscle in postmenopausal women. AB - The accretion of fat within skeletal muscle has been associated with metabolic abnormalities. Leptin increases muscle fatty acid oxidation and triglyceride hydrolysis. Therefore, leptin concentrations may affect muscle fat content. The objective of this study was to determine if serum leptin concentrations were associated with im lipid content, as reflected in the mid-thigh low-density skeletal muscle area (LDMA). In addition, we evaluated whether hormone replacement therapy (HRT) or ethnicity affected this relationship. Our study population consisted of 80 postmenopausal women aged 45-55 yr, (72 Caucasian and 8 African-American). Both HRT users (n = 50) and nonusers (n = 30) were recruited. Total fat mass was estimated using total body dual-energy x-ray absorptiometry. Fat and muscle areas at the mid-thigh were measured using computed tomography scanning. Results showed that, after adjusting for total fat mass, higher-density muscle area, and ethnicity, higher serum leptin concentration was associated with lower LDMA (P < 0.05). African-American women had greater LDMA than Caucasian women, after controlling for leptin concentration (P < 0.05). Use of HRT did not significantly influence LDMA. These results support the hypothesis that leptin decreases skeletal muscle lipid content, promoting lipid oxidation. PMID- 12629100 TI - Triglyceride-rich lipoproteins are associated with hypertension in preeclampsia. AB - Disorders of the lipoprotein metabolism are a major cause of endothelial dysfunction that may result in hypertension and proteinuria, clinical hallmarks of preeclampsia (PE). Lipoproteins and low-density lipoprotein (LDL) subfractions were investigated in 15 women with severe PE and compared with 23 women with a normal course of pregnancy. Compared with normal pregnancy, in PE apolipoprotein (apo)B in very low-density lipoprotein was increased by 76% (P = 0.008), and the triglyceride content of intermediate dense lipoproteins (IDL) was increased by 51% (P < 0.001); cholesterol and apoB in LDL were decreased by 26% (P = 0.005) and 23% (P = 0.016), respectively. Although not significant, the LDL profile was dominated by the most buoyant LDL-1. ApoB in the most dense LDL (dLDL), namely LDL-5 and LDL-6, was significantly decreased by 49% (P < 0.001) and 55% (P < 0.001), respectively. Diastolic blood pressure was positively correlated with the triglyceride content of IDL (r = 6.31; P < 0.001 and r = 0.352; P = 0.033 by partial correlation controlling for the presence or absence of PE) and negatively correlated with the concentration of apoB in dLDL (r = -0.500; P = 0.002). In addition, IDL triglycerides correlated negatively with infant birth weight percentile (r = -0.373; P = 0.027) and positively with proteinuria (r = 0.430; P = 0.014). Low birth weight was associated with high IDL triglycerides and low rather than high concentrations of dLDL. Triglyceride-rich remnants are known to cause endothelial dysfunction. Because the triglyceride content of IDL was positively correlated with elevated blood pressure and proteinuria, triglyceride rich remnant lipoproteins might contribute to the pathophysiology of PE. PMID- 12629102 TI - Raloxifene lowers serum calcium and markers of bone turnover in postmenopausal women with primary hyperparathyroidism. AB - Estrogen replacement therapy (ERT) decreases total serum calcium by about 0.5 mg/dl in postmenopausal women with primary hyperparathyroidism (PHPT). We investigated the ability of raloxifene, which has skeletal antiresorptive properties similar to those of ERT, to decrease serum calcium concentrations and markers of bone turnover in PHPT. Eighteen postmenopausal women with asymptomatic PHPT were randomized to 8 wk of raloxifene (60 mg/d) or placebo, followed by a 4 wk washout. At baseline, the groups were well matched. The calcium concentration decreased significantly by 8 wk of raloxifene administration (10.8 +/- 0.2 to 10.4 +/- 0.2 mg/dl; P < 0.05), as did markers of bone resorption and formation [osteocalcin, 11.4 +/- 1.6 to 9.9 +/- 1.6 nmol/liter (P < 0.05); serum N telopeptide, 21.2 +/- 3.4 to 17.3 +/- 2.8 nmol bone collagen equivalents/liter (P < 0.05)]. Four weeks after raloxifene was discontinued, indices were indistinguishable from baseline. Raloxifene administration did not affect serum PTH, 1,25-dihydroxyvitamin D, total alkaline phosphatase, or urinary calcium excretion. Calcium and bone marker changes were therefore similar to those observed with ERT in PHPT. This short-term study suggests that raloxifene may be a useful approach to the treatment of postmenopausal women with mild PHPT. PMID- 12629101 TI - The male contraceptive regimen of testosterone and levonorgestrel significantly increases lean mass in healthy young men in 4 weeks, but attenuates a decrease in fat mass induced by testosterone alone. AB - In hypogonadal men, testosterone (T) in replacement dosages is known to increase fat-free mass (lean mass) and decrease fat mass. In young eugonadal men, similar dosages of T increase lean mass, but much higher dosages of T are required to decrease total body fat mass. Current T-based male hormonal contraceptive regimens include a second agent, such as a progestin, to maximize inhibition of pituitary gonadotropins and improve efficacy. To study the effect of such combinations on body composition, we randomized healthy, young, eugonadal men into four combinations of exogenous T and the progestin, levonorgestrel (LNG): 1) 100 mg T enanthate, im, weekly plus 125 micro g LNG, orally, daily (T+LNG); 2) T plus placebo LNG (T alone); 3) placebo T plus LNG (LNG alone); and 4) placebo T plus placebo LNG (placebo). We then analyzed body composition by dual energy x ray absorptiometry after 4 and 8 wk of treatment. T+LNG significantly increased total lean mass after 4 and 8 wk of treatment (3.5 +/- 0.9% and 4.2 +/- 1.2%, respectively; P < 0.05) and truncal lean mass after 4 and 8 wk of treatment (4.7 +/- 0.9% and 5.0 +/- 0.9%, respectively; P < 0.05) compared with baseline and placebo. T alone also increased total and truncal lean mass significantly compared with placebo after 4 wk of treatment, but not compared with baseline (3.3 +/- 1.4% and 3.2 +/- 2.3%, respectively; P < 0.05 vs. placebo), suggesting an additive effect of T and LNG to increase lean mass. Fat mass significantly decreased in the abdomen in men administered T alone compared with LNG alone ( 4.9 +/- 2.8%; P < 0.05). Fat mass significantly increased in the abdomen with LNG alone (4.1 +/- 1.0%; P < 0.05) compared with baseline and was unchanged with the combination of T+LNG, suggesting that LNG attenuates the decrease in fat mass seen with T alone. There was no change in weight or body mass index in any group during the study. This study shows that in young eugonadal men 1) T alone rapidly increases lean mass and decreases fat mass in 4-8 wk; 2) T+LNG rapidly increases lean mass, but has no effect on fat mass; and 3) LNG alone increases fat mass. The favorable profile on body composition by T is, therefore, partially attenuated by the progestin, LNG. These findings suggest that androgen-based male hormonal contraceptives might have favorable effects on body composition. The impact of these changes on cardiovascular risk in normal men needs further study. PMID- 12629103 TI - Can some growth hormone (GH)-deficient children benefit from combined therapy with gonadotropin-releasing hormone analogs and GH? Results of a retrospective study. AB - Recombinant GH (rGH) treatment does not invariably correct height deficits in GH deficient children once puberty has begun. The addition of GnRH analogs (GnRHa) to delay puberty has been advocated, but published results are few and sometimes conflicting. We retrospectively compared GH-deficient children treated with rGH and GnRHa for at least 1 yr after entering puberty and having attained their final height (n = 23) with a matched control group treated only with rGH. Overall, combined therapy did not significantly increase final height relative to rGH alone. However, the shortest girls at the onset of puberty (<25th percentile) benefited more than the tallest (>75th percentile) in both final height relative to predicted height and pubertal catch-up growth. In the control group, patients having experienced intrauterine growth retardation (IUGR) attained a lower mean final height than patients without IUGR (difference significant in boys, but not in girls). In the combined therapy group, IUGR did not affect the final height of either sex. Our results suggest that two populations might benefit most from combined GnRHa and rGH therapy: girls particularly short at the onset of puberty and patients who had experienced IUGR. Further prospective studies are required to confirm these preliminary hypothesis. PMID- 12629104 TI - Alpha(2B)-adrenergic receptor deletion polymorphism associates with autonomic nervous system activity in young healthy Japanese. AB - Several polymorphisms of genes involved in autonomic nervous system (ANS) function have been reported to affect metabolic regulation. We have investigated the association of an alpha(2B)-adrenergic receptor (alpha(2B)AR) three-amino acid deletion polymorphism with ANS activity in young healthy subjects by means of electrocardiogram R-R interval power spectral analysis. Three hundred eighty one young healthy Japanese males (mean +/- SE, 20.6 +/- 0.1 yr) were studied. One hundred sixty-eight (44.1%) were homozygotes of Long allele (Long/Long), 162 (42.5%) were heterozygotes (Long/Short), and 51 (13.4%) were homozygotes of Short allele (Short/Short). The allele frequency of Short allele was 0.35. No significant differences were observed in any of the characteristics investigated: body mass index, plasma glucose, plasma insulin, or family history of diabetes and obesity. In R-R spectral analysis of heart rate variability, carriers of Short/Short had significantly greater low frequency and very low frequency than Long/Long, as well as a higher sympathetic nervous system index. These findings suggest that the alpha(2B)AR deletion polymorphism might result in metabolic disorder by altering ANS function. PMID- 12629105 TI - Diminished interleukin-6 response to proinflammatory challenge in men and women after intravenous cocaine administration. AB - Cocaine abuse is associated with increased rates of infections, including human immunodeficiency virus, and cocaine has immunomodulatory effects in experimental animal and cellular models. When challenged by antigens, tissues release cytokine polypeptides that signal a complex balance of cellular and humoral immune responses. Placement of indwelling venous catheters also leads to surrounding tissue inflammation, mediated partially by local production and release of the proinflammatory cytokine, IL-6. Thus, catheter placement provides a model for examination of cocaine's immunological effects. Thirty healthy men and women with a history of cocaine use participated in this study of neuroendocrine and immunological responses to iv injection of 0.4 mg/kg cocaine or saline placebo. After injection, blood samples were collected from the antecubital vein of the opposite arm via an indwelling venous catheter at 2, 4, 8, 12, 16, 20, 30, 40, 60, 80, 120, 180, and 240 min. Cocaine, ACTH, cortisol, and dehydroepiandrosterone concentrations peaked at 8, 12, 40, and 20 min, respectively. Stimulation of IL-6 at 240 min was markedly reduced in subjects receiving cocaine compared with subjects receiving placebo (3.85 +/- 0.49 vs. 11.64 +/- 2.21 pg/ml; P = 0.0019, by two-tailed t test). Gender and menstrual cycle phase did not significantly influence most endocrine or IL-6 measures, although the small number of subjects limits the power of these comparisons. Because cocaine stimulates the hypothalamic-pituitary-adrenal axis, IL-6 suppression may be a consequence of corticosteroid release. Cocaine-induced suppression of proinflammatory IL-6 may mediate impaired host defenses to infections. PMID- 12629106 TI - Desensitization of Gs-coupled receptor signaling by constitutively active mutants of the human lutropin/choriogonadotropin receptor. AB - Activating mutations of the human lutropin/choriogonadotropin receptor (hLHR), a Gs-coupled receptor, have been identified in young boys with gonadotropin independent precocious puberty (testotoxicosis). The properties of these mutants have typically been characterized in heterologous cells transfected with recombinant mutant receptor and compared with those expressing wild-type (wt) receptor. The affected individuals, however, are heterozygous and, therefore, express wt receptor in addition to the mutant receptor. The present studies were undertaken to determine what effects, if any, coexpression of a constitutively active hLHR might have on hLHR(wt). HEK 293 cells were cotransfected with hLHR(wt) and hLHR(L457R), a mutant that we have previously shown to be both constitutively active and unresponsive to further hormonal stimulation as determined in both intact cells and isolated membranes. When coexpressed at submaximal concentrations, L457R does not decrease the cell surface expression of hLHR(wt). Coexpression of L457R, however, causes an attenuation of human choriogonadotropin-stimulated cAMP production by hLHR(wt). We show that this attenuation is caused by an activation of the phosphodiesterase (PDE)4D3. Additional experiments demonstrate that the coexpression of L457R with the human beta(2)-adrenergic receptor causes an attenuation of isoproterenol-stimulated cAMP and that other activating mutations of the hLHR also induce PDE activation. Taken together, these data demonstrate that the activation of PDE is a compensatory mechanism common to hLHR constitutively active mutants and that cellular responses to agonists that stimulate Gs-coupled receptors may be blunted in tissues expressing these activating mutants. This novel desensitizing effect of constitutively active hLHRs on hormone-stimulated cAMP production has not been noticed before and would typically not be detected because of the routine inclusion of PDE inhibitors in experiments determining cAMP accumulation. Importantly, however, this mechanism of desensitization would be expected to occur in a physiological context in which PDE inhibitors are not present and thus may influence hormonal signaling in cells expressing the activating hLHR mutant. PMID- 12629107 TI - Leptin stimulates the activity of the system A amino acid transporter in human placental villous fragments. AB - The activity and expression of placental nutrient transporters are primary determinants for the supply of nutrients to the fetus, and these nutrients in turn regulate fetal growth. We developed an experimental system to assess amino acid uptake in single primary villous fragments to study hormonal regulation of the amino acid transporter system A in term human placenta. Validation of the method, using electron microscopy and studies of hormone production, indicated that fragments maintained ultrastructural and functional integrity for at least 3 h. The activity of system A was measured as the Na(+)-dependent uptake of methylaminoisobutyric acid (MeAIB), and the effect of 1 h incubation in various hormones was investigated. Uptake of MeAIB into villous fragments in the presence of Na(+) was linear up to at least 30 min. Insulin (300 ng/ml, n = 14) increased system A activity by 56% (P < 0.05). This effect was also present at insulin concentrations in the physiological range (+47% at 0.6 ng/ml, n = 10, P < 0.05). Leptin (500 ng/ml, n = 14) increased Na(+)-dependent MeAIB uptake by 37% (P < 0.05). System A activity increased in a concentration-dependent fashion in response to leptin (n = 10). However, neither epidermal GF (600 ng/ml), cortisol (340 ng/ml), nor GH (500 ng/ml) altered system A activity significantly (n = 14). We conclude that primary single isolated villous fragments can be used in studies of hormonal regulation of nutrient uptake into the syncytiotrophoblast. These data suggest that leptin regulates system A, a key amino acid transporter. PMID- 12629108 TI - Additional phenotypic abnormalities with presence of cysts within the empty thyroid area in patients with congenital hypothyroidism with thyroid dysgenesis. AB - Congenital hypothyroidism (CH) is most frequently caused by thyroid developmental abnormalities, and it has recently been shown to have a familial component with members affected by either CH or asymptomatic thyroid developmental abnormalities. The pathogenesis of the disease is unknown, but it seems possible that a common genetic mechanism underlies these heterogeneous phenotypic expressions. Associations among these anomalies in the same individuals have occasionally been described. The aim of this study was to investigate whether cysts of the thyroglossal duct could be shown by ultrasonography in patients with CH caused by thyroid dysgenesis. Children with CH (n = 57) who were diagnosed by newborn TSH screening were prospectively evaluated by ultrasonography at the age of 10.5 +/- 4.5 yr. The etiology of CH (ectopic thyroid tissue, n = 42; athyreosis, n = 15) was established before treatment initiation on the basis of thyroid radioiodine scanning and the absence of any thyroid tissue in the normal location confirmed by ultrasonography. Cysts were found in 39 patients (68% of cases) with either ectopic thyroid tissue (n = 29) or athyreosis (n = 10). All cysts were located in the empty thyroid area in the left (57%) or right (43%) side and were mostly closer to the midline. Patients had either a single cyst (n = 16 patients) or multiple cysts (n = 23 patients). The cysts were bilateral in 17 of the 39 patients. Most of them were vertically oval or round, with a size ranging in diameter from 2-21 mm (mean, 3.5 +/- 2). In conclusion, the presence of cysts within the empty thyroid area in 68% of patients with CH due to thyroid dysgenesis is a novel observation that is part of the developmental anomaly of this disease. Several explanations can be put forward to explain the presence of these cysts. They might be due to the persistence of the ultimobranchial bodies as a cystic structure or part of the thyroid-forming material, which may migrate along the normal pathway of the usual course of the thyroglossal duct, giving rise to cell residues within the empty thyroid area. PMID- 12629109 TI - Blood pressure, serum lipids, fasting insulin, and adrenal hormones in 12-year old children born with maternal preeclampsia. AB - Women with prior preeclamptic pregnancies have an increased risk for metabolic syndrome and cardiovascular diseases. Maternal preeclampsia has been associated with elevated blood pressure (BP) in offspring during childhood. The aim of our study was to determine whether elevated BP pressure and metabolic changes, such as dyslipidemia, insulin resistance, and increased adrenal hormonal activity, are found in 12-yr-old children of preeclamptic mothers. Sixty children born after preeclamptic pregnancy (PRE) and 60 matched control subjects born after normotensive pregnancy (non-PRE) were studied at the age of 12 yr. The case control pairs were matched for sex, gestational age (+/-1 wk), and size at birth. We measured BP and concentrations of blood glucose, serum fasting insulin, total and high density lipoprotein cholesterol, triglycerides, cortisol, dehydroepiandrosterone sulfate, and plasma epinephrine (E) and norepinephrine (NE). Low density lipoprotein cholesterol was calculated according to the Friedewald-Fredrickson formula. The PRE children had significantly higher mean systolic (116.4 vs. 113.2 mm Hg; P = 0.021) and diastolic (73.9 vs. 70.3 mm Hg; P = 0.022) BP than the non-PRE children, even when adjusted by current weight and height. At 12 yr of age, systolic BP values correlated inversely with birth weight (r = -0.459; P < 0.001) and length SD scores (r = -0.429; P = 0.001) in the PRE children. The mean concentrations of serum total, low density lipoprotein, and high density lipoprotein cholesterol; triglycerides; fasting insulin; blood glucose; serum cortisol; and dehydroepiandrosterone sulfate did not differ between the PRE and non-PRE groups. However, the mean plasma E concentration was higher in the PRE than in the non-PRE children (0.32 vs. 0.28 nmol/liter; P = 0.042), whereas the mean NE concentration did not differ between these two groups. In conclusion, 12-yr-old children born with maternal preeclampsia had elevated systolic and diastolic BPs and slightly increased E levels in the circulation. It is not known whether these changes are caused by genetic factors or by preeclampsia itself. PMID- 12629110 TI - Validation and calibration of the Kabi Pharmacia International Growth Study prediction model for children with idiopathic growth hormone deficiency. AB - In 1999 a model was published for prediction of growth in children with idiopathic GH deficiency (IGHD) during GH therapy, derived using data from the Kabi Pharmacia International Growth Study (KIGS) database (Pharmacia ?|[amp ]?| Upjohn, Inc., International Growth Database). We validated and calibrated this KIGS model for growth in the first year of GH therapy using data from 136 Dutch children with IGHD. Observed vs. predicted outcomes were plotted, and the fitted regression line was significantly different from the line of identity (P = 0.03). It appeared that the predictions were too extreme: relatively low predictions were too low, relatively high predictions were too high. This is a well known phenomenon in the context of prediction models, called overoptimism. For valid application to other data the KIGS predictions should be calibrated. Calibrated predictions are obtained using Y(cal) = Y(orig) + (2.153 - 0.192 x Y(orig)), where Y(cal) is the calibrated prediction, and Y(orig) is the KIGS prediction. The calibrated prediction will be higher than the original KIGS prediction when the original prediction is less than 11.2 cm/yr and will be lower otherwise. The variability of the prediction errors of the calibrated predictions was positively related to the value of the prediction (P < 0.001), described by the equation SD(pred err) = -1.017 + 0.286 x Y(cal). Our calibrated model will give better predictions for children with IGHD fulfilling the same criteria. PMID- 12629111 TI - Induction of postprandial inflammatory response in adult onset growth hormone deficiency is related to plasma remnant-like particle-cholesterol concentration. AB - Increased cardiovascular mortality due to premature atherosclerosis is a clinical feature in the adult-onset GH deficiency (AGHD) syndrome. Inflammation is a key feature in atherogenesis and may be triggered by postprandial lipoprotein remnants. We hypothesized that increased postprandial lipoprotein remnant levels in AGHD may be associated with an inflammatory response. In this case-control study, 10 AGHD patients [6 males and 4 females; age, 48 +/- 9 yr; body mass index (BMI), 26.9 +/- 2.6 kg/m(2)] and 10 healthy control subjects (matched for age, BMI, gender, baseline lipid levels, and apolipoprotein E genotype) were included. They all ingested an oral fat load. Fasting and postprandial levels of plasma remnant-like particle-cholesterol (RLP-C; 0.31 +/- 0.13 mmol/liter and 4.14 +/- 1.37 mmol/liter.h in GHD; 0.18 +/- 0.06 mmol/liter and 2.56 +/- 1.02 mmol/liter.h in controls, respectively) were significantly increased in AGHD patients compared with control subjects. The median inflammatory cytokines, IL-6 and TNF-alpha, were higher in the fasting [3.9 (range, 3.1-11.9) pg/ml and 6.8 (range, 2.5-27.6) pg/ml, respectively] and postprandial [151.7 (range, 87.0-294.3) pg/ml.24 h and 289.9 (range, 87.5-617.6) pg/ml.24 h, respectively] states in AGHD than in controls [fasting, 0.9 (range, 0.2-5.2) pg/ml and 2.8 (range, 2.5-5.7) pg/ml; and postprandial, 54.5 (range, 11.50-126.5) pg/ml.24 h and 118.3 (range, 81.2-243.1) pg/ml.24 h, respectively]. In addition, postprandial profile of RLP-C and IL-6 in AGHD and in the total group were significantly associated (r(2) = 0.44, P < 0.05; and r(2) = 0.38, P < 0.01, respectively). In conclusion, the increased postprandial RLP-C level in GHD is associated with an inflammatory response that may result in increased susceptibility for premature atherosclerosis. PMID- 12629112 TI - Serum leptin levels as a marker for a syndrome X-like condition in wild baboons. AB - We measured serum leptin levels in two groupings of wild male baboons, one with access to abundant quantities of food from gardens and garbage dumps near human habitations (Garbage; n = 11) and one without access (No Garbage; n = 10). A Garbage subgroup had high leptin levels (Garbage HL), whereas the rest of the Garbage group had low leptin levels (Garbage LL) similar to those in the No Garbage group. The Garbage HL individuals were obese, with higher mass, body mass index, and leptin to mass ratios; were insulin to resistant, with elevations in serum insulin, glucose, and insulin to glucose ratios; and were hyperlipidemic. This syndrome X-like condition occurred only in the Garbage HL subset. The Garbage LL subset did not differ from the No Garbage individuals in mass, body mass index, leptin to mass ratio, insulin, glucose, or insulin to glucose ratios. The highest cholesterol levels, however, occurred in the Garbage LL individuals, suggesting that susceptibility to hyperlipidemia is distinguishable from susceptibility to obesity and insulin resistance. The differences were not explained by age or social status. These results show that a subgroup of wild baboons is susceptible to developing obesity and insulin resistance and that this susceptibility is not related to age or social rank. PMID- 12629113 TI - A novel nonsense mutation in the Pit-1 gene: evidence for a gene dosage effect. AB - The POU transcription factor Pit-1 functions in the development of somatotrophs, lactotrophs, and thyrotrophs of the anterior pituitary gland. It also plays a role in cell-specific gene expression and regulation of the gene products from these cell types, GH, prolactin, and TSH, respectively. In the present report we studied a patient with severe growth failure. Provocative studies revealed undetectable GH, prolactin, and TSH levels, and her pituitary gland was hypoplastic on magnetic resonance imaging. She had a novel homozygous nonsense mutation in the 3' end of the first alpha-helix of the POU-specific domain of the Pit-1 gene. This mutation results in a truncated protein with loss of most of the Pit-1 DNA-binding domains. Interestingly, her parents, who each have one mutant allele, have evidence of mild endocrine dysfunction. Thus, two normal copies of the Pit-1 gene appear necessary for full Pit-1 gene function. PMID- 12629115 TI - Normal insulin-stimulated endothelial function and impaired insulin-stimulated muscle glucose uptake in young adults with low birth weight. AB - Low birth weight has been linked to insulin resistance and cardiovascular disease. We hypothesized that insulin sensitivity of both muscle and vascular tissues were impaired in young men with low birth weight. Blood flow was measured by venous occlusion plethysmography during dose-response studies of acetylcholine and sodium nitroprusside in the forearm of fourteen 21-yr-old men with low birth weight and 16 controls of normal birth weight. Glucose uptake was measured during intraarterial insulin infusion. Dose-response studies were repeated during insulin infusion. The maximal blood flow during acetylcholine infusion was 14.1 +/- 2.7 and 14.4 +/- 2.1 [ml x (100 ml forearm)(-1) x min(-1)] in low and normal birth weight subjects, respectively. Insulin coinfusion increased acetylcholine stimulated flow in both groups: 18.0 +/- 3.1 vs. 17.9 +/- 3.1 [ml x (100 ml forearm)(-1) x min(-1)], NS. Insulin infusion increased glucose uptake significantly in the normal birth weight group, compared with the low birth weight group: 0.40 +/- 0.09 to 1.00 +/- 0.16 vs. 0.44 +/- 0.09 to 0.59 +/- 0.1 [ micro mol glucose x (100 ml forearm)(-1) x min(-1)], P = 0.04. Young men with low birth weight have normal insulin-stimulated endothelial function and impaired insulin-stimulated forearm glucose uptake. Thus, endothelial dysfunction does not necessarily coexist with metabolic alterations in subjects with low birth weight. PMID- 12629114 TI - Analysis of microsatellite instability in sporadic parathyroid adenomas. AB - Microsatellite instability (MSI) is the form of genomic instability associated with defective DNA mismatch repair (MMR) in human tumorigenesis. Recent reports have suggested a role for MSI in the pathogenesis of sporadic parathyroid adenomas. However, because of their small sample sizes and/or lack of systematic analysis of genome-wide MSI, these studies have not provided conclusive evidence that MMR defects are a common occurrence in parathyroid neoplasia. To further investigate whether MSI plays an important role in parathyroid tumorigenesis, we analyzed 49 sporadic parathyroid adenomas for MSI using a panel of 5 microsatellite DNA markers that has been recommended for sensitive detection of MSI by the NCI Workshop and validated in other tumor types. These microsatellite loci were amplified by PCR using fluorescent-labeled primers from the 49 samples of template tumor DNA and matching normal DNA isolated from the same patients' peripheral blood leukocytes. None of the 49 tumors showed evidence of MSI at any of the analyzed loci of the NCI marker panel. These observations strongly suggest that defective DNA MMR plays a minor role, if any, in the pathogenesis of sporadic parathyroid adenomas. PMID- 12629116 TI - Association of resistin gene 3'-untranslated region +62G-->A polymorphism with type 2 diabetes and hypertension in a Chinese population. AB - Resistin, a recently discovered polypeptide, antagonizes insulin action and may play a part in the pathogenesis of insulin resistance. This study investigates whether resistin gene polymorphism can be associated with type 2 diabetes. We studied 1102 Chinese type 2 diabetes patients and 743 subjects without diabetes. The resistin 3'-untranslated region (UTR) +62G-->A polymorphism was determined by PCR. Type 2 diabetes subjects had a lower frequency of resistin gene 3'UTR +62A allele (GG:GA/AA, 83.5%:16.5%) than the controls (GG:GA/AA, 75.1%:24.9%; odds ratio, 1.524; 95% confidence interval, 1.268-1.831; P < 0.001). Unexpectedly, diabetic patients with the GG genotype had a higher prevalence of hypertension (GG:GA/AA, 49.8%:36.2%; odds ratio, 1.375; 95% confidence interval, 1.116-1.693; P = 0.001). Logistic regression analysis confirmed that the resistin gene 3'UTR +62G-->A polymorphism acts as an independent contributing factor to type 2 diabetes and hypertension. The mean systolic and diastolic blood pressure levels in diabetic subjects with the GG genotype (144 +/- 21/87 +/- 13 mm Hg) were significantly higher than those in subjects with GA/AA variants (139 +/- 21/84 +/ 14 mm Hg; P = 0.004 and P = 0.002, respectively). Multiple linear regression analysis showed resistin gene polymorphism to be an independent factor associated with systolic and diastolic blood pressures in type 2 diabetes patients. These findings suggest that resistin may play a role in the pathogenesis of type 2 diabetes and insulin resistance-related hypertension. PMID- 12629117 TI - Increased insulin sensitivity is associated with reduced insulin and glucagon secretion and increased insulin clearance in man. AB - Insulin secretion is increased in insulin resistance. In this study, we examined whether high insulin sensitivity results in low insulin secretion. Twelve male master athletes [age 25.6 +/- 4.1 (mean +/- SD) yr] and seven male sedentary students (age 25.0 +/- 2.0 yr) underwent a hyperinsulinemic, euglycemic clamp and a glucose-dependent arginine stimulation test. Athletes had high insulin sensitivity [230 +/- 18 vs. 92 +/- 12 (nmol glucose/kg.min)/(pmol insulin/liter), P < 0.001] and low insulin response to arginine (at fasting glucose 135 +/- 22 vs. 394 +/- 60 pmol/liter, P < 0.001), which resulted in unaltered disposition index (32.8 +/- 3.8 vs. 33.5 +/- 3.3 micro mol glucose/kg.min, NS). Also, the C peptide response to arginine was reduced (at fasting glucose 0.71 +/- 0.09 vs. 0.89 +/- 0.09 nmol/liter, P = 0.034). However, the C-peptide reduction was not as large as the insulin reduction yielding increased disposition index in athletes when calculated from C-peptide data (184 +/- 9 vs. 76 +/- 11 micro mol glucose/kg.min, P < 0.001). This difference is explained by increased insulin clearance among the athletes during the first 5 min after arginine (81.1% +/- 1.8% vs. 53.6% +/- 4.7%, P < 0.001). Also, the glucagon response to arginine was reduced in the athletes (58.8 +/- 6.7 vs. 90.1 +/- 9.9 ng/liter at fasting glucose, P = 0.009). We conclude that high insulin sensitivity results in low islet hormone secretion and increased insulin clearance. PMID- 12629118 TI - Glucose transport and phosphorylation in skeletal muscle in obesity: insight from a muscle-specific positron emission tomography model. AB - A controversial area in understanding the contribution of obesity to skeletal muscle insulin resistance is the distribution of control of glucose metabolism across proximal steps of glucose delivery, trans-membrane transport, and intracellular trapping via phosphorylation. Dynamic positron emission tomography (PET) imaging of skeletal muscle [(18)F]2-deoxy-2-D-glucose ((18)F-FDG) uptake provides an in vivo method for assessment of these steps in humans. In the current study we have examined the application of a four-compartment skeletal muscle-specific model for assessment of (18)F-FDG metabolism that takes interstitial (18)F-FDG kinetics into account and compared this to the classic three-compartment model in lean and obese volunteers. We assessed the effects of insulin infusions at three rates (0, 40, and 120 mU/m(2).min). In comparison with the classic model, the skeletal muscle-specific model reveals more clearly definable effects of insulin on transmembrane glucose transport and an impairment of this response in obesity. Compared with the classic model for assessment of (18)F-FDG metabolism, both the skeletal muscle-specific and the classic model indicate that, with respect to distribution of control, glucose phosphorylation has an important effect at low to moderate levels of insulin stimulation in both lean and obese subjects. PMID- 12629119 TI - Postpartum thyroiditis is associated with fluctuations in transforming growth factor-beta1 serum levels. AB - Postpartum thyroiditis (PPT) is characterized by a rapid evolution and recovery of euthyroidism. Therefore, it can represent a good model to study early cytokine fluctuations in autoimmune thyroid diseases. TGFbeta1 is an immunosuppressive cytokine, as it inhibits T and B cell proliferation, natural killer cell cytotoxic activity, and the generation of T cell cytotoxicity. The aim of this study was to assess serum concentrations of TGFbeta1 during pregnancy and to study possible serum fluctuations of this cytokine during the different phases of PPT. Thyroid biochemical pattern, antithyroid autoantibodies (ATA), and total and active TGFbeta1 (aTGFbeta1) serum concentrations were evaluated in 63 pregnant women. Thirty-four of them were ATA(+), and 29 were ATA(-). Twenty of the 34 ATA(+) women were followed in the postpartum year. Nine of these 20 women developed PPT; 11 remained euthyroid. All of the PPT women became euthyroid during the follow-up. Our results showed 1) detectable serum levels of aTGFbeta1 in 50% of ATA(+) pregnant women, suggesting that the presence of autoantibodies may characterize a favorable condition for TGFbeta1 activation; and 2) decreased total TGFbeta1 and increased aTGFbeta1 serum levels during the active phase of PPT in ATA(+) women. This seems to suggest that inflammation may be responsible for TGFbeta1 activation and autoantibody increase because of antigen release. Although further studies of women with persistent hypothyroidism after the postpartum year are needed, the possibility that the enhanced activation of TGFbeta1 may contribute to resolution of thyroid inflammation postpartum cannot be excluded. PMID- 12629120 TI - Endotoxin stimulates leptin in the human and nonhuman primate. AB - Leptin, which plays a key role in regulating energy homeostasis, may also modulate the inflammatory response. An inflammatory challenge with endotoxin has been shown to stimulate leptin release in the rodent. This finding has not been reproduced in humans or in nonhuman primates, although leptin levels have been reported to increase in septic patients. We have therefore examined the effects of endotoxin injection on plasma leptin levels in nine ovariectomized monkeys and four postmenopausal women. In an initial study in five monkeys, mean leptin levels did not increase during the first 5 h after endotoxin treatment, but did increase significantly from 6.4 +/- 2.1 ng/ml at baseline to 12.3 +/- 4.4 ng/ml at 24 h (P = 0.043). In a second study, a significant increase in leptin over time was noted after endotoxin treatment (P < 0.001); leptin release during the 16- to 24-h period after endotoxin injection was 48% higher than during the control period (P = 0.043). A similar stimulatory effect of endotoxin on leptin was observed when monkeys received estradiol replacement. In a third study, repeated injections of endotoxin over a 3-d period stimulated IL-6, ACTH, cortisol, and leptin release (P < 0.001). Leptin increased during the first day of treatment in all animals, but only monkeys with baseline plasma leptin levels greater than 10 ng/ml exhibited a sustained increase in leptin throughout the 3-d period. There was a significant correlation (r = 0.81; P = 0.008) between the mean baseline leptin level and the percent increase in leptin over baseline on the last day of treatment. In the human subjects, plasma leptin concentrations did not change significantly during the 7-h period after endotoxin injection. However, leptin increased in all four women from a mean baseline of 8.34 +/- 3.1 to 13.1 +/- 4.3 ng/ml 24 h after endotoxin (P = 0.038). In summary, endotoxin stimulates the release of leptin into peripheral blood in the human and nonhuman primate, but the time course is different from that reported in the rodent. These results are consistent with previous reports of increased blood leptin levels in patients with sepsis. The significance of these findings and the potential role of leptin in modulating the response to inflammation in the human require further study. PMID- 12629121 TI - A genome-wide scan for autoimmune thyroiditis in the Old Order Amish: replication of genetic linkage on chromosome 5q11.2-q14.3. AB - Autoimmune thyroiditis (AITD) is a common disorder characterized by circulating antibodies to epitopes of thyroid tissue and hypothyroidism (Hashimoto's thyroiditis or AITD-hypothyroidism), although many subjects with AITD are euthyroid. Current evidence suggests that AITD is familial and polygenic. We studied AITD in a homogeneous founder Caucasian population, the Old Order Amish of Lancaster County, Pennsylvania. We found autoimmune thyroiditis, defined by the presence of circulating antimicrosomal antibodies, to be relatively common in the Amish, with a prevalence of 22.7%. The prevalence of AITD-hypothyroidism was 9.2%. We performed a genome-wide linkage analysis with 373 short tandem repeat markers in 445 subjects from 29 families. We observed suggestive evidence of linkage of AITD to a locus on chromosome 5q11.2-q14.3 (LOD, 2.30; P = 0.0006 at 94 cM; closest marker, D5S428), a region that was previously reported to be linked to AITD-hypothyroidism in a Japanese study. AITD-hypothyroidism showed a more modest linkage peak to the same region (LOD, 1.46; P = 0.005). Possible linkage (nominal P < 0.01) to autoimmune thyroiditis and/or AITD-hypothyroidism was also detected in five other regions. We conclude that a gene on chromosome 5q11.2-q14.3 is likely to contribute to susceptibility to AITD in the Amish. PMID- 12629122 TI - Nongenomic effects of aldosterone on human renal cells. AB - The development of chronic renal insufficiency may be partially mediated by the nongenomic action of aldosterone. Here we investigate whether aldosterone could evoke a nongenomic action in primary cultures of human renal cells. Intracellular Ca(2+) ([Ca(2+)](i)) and cAMP were measured in human mesangial cells (MC), glomerular visceral epithelial cells (GVEC), and proximal and distal tubular epithelial cells (Ptec and Dtec) in the presence of aldosterone (10-100 nmol/liter) by fura-2 fluorescence and RIA, respectively. In MC, Ptec, and Dtec, aldosterone increased [Ca(2+)](i) within 1 min, whereas in GVEC, only a minor effect was found. Preincubation of cells with spironolactone did not blunt this effect. Hydrocortisone, used at a concentration 100-fold higher than that of aldosterone, did not affect [Ca(2+)](i.) In MC, Ptec, and Dtec, a dose-dependent increase ( approximately 1.3- to 1.5-fold) in intracellular cAMP levels was found. These data demonstrate a nongenomic action of aldosterone in human MC, Ptec, and Dtec. As these effects occur at concentrations close to free plasma aldosterone levels in man, they may be of physiological relevance and may contribute to renal injury. PMID- 12629123 TI - A comparison of low-dose and standard-dose oral estrogen on forearm endothelial function in early postmenopausal women. AB - We investigated the effects of low-dose estrogen plus progestin on endothelial function. Postmenopausal women received daily doses of conjugated equine estrogen (CEE, 0.625 mg) plus medroxyprogesterone acetate (MPA, 2.5 mg) (standard-dose group, n = 18), CEE (0.3 mg) plus MPA (2.5 mg) (low-dose group, n = 18), or no treatment (control group, n = 15) for 3 months. Serum concentrations of lipids and malondialdehyde (MDA)-modified low-density lipoprotein (LDL) were measured. Forearm blood flow (FBF) during reactive hyperemia and after sublingual nitroglycerin administration was measured by strain-gauge plethysmography. Decreases in serum concentrations of LDL cholesterol and MDA-modified LDL and increases in high-density lipoprotein cholesterol and nitrite/nitrate were observed in both treatment groups. After 3 months of treatment, similar increases in the maximal FBF response during reactive hyperemia were observed in both treatment groups (standard-dose group, from 35.8 +/- 3.0 to 47.5 +/- 2.8 ml/min per 100 ml tissue; and low-dose group, from 35.2 +/- 2.2 to 46.8 +/- 3.4 ml/min per 100 ml tissue, P < 0.01). FBF levels in the control group were unchanged. Treatment did not affect nitroglycerin-induced dilation. The incidences of vaginal bleeding and breast tenderness were lower with the low-dose group than with the standard-dose group. Low-dose CEE plus MPA augments endothelial function in forearm resistance arteries and decreased MDA-modified LDL levels similarly to standard doses of CEE plus MPA, with fewer side effects. PMID- 12629124 TI - Calcitonin-specific transcription and splicing targets gene-directed enzyme prodrug therapy to medullary thyroid carcinoma cells. AB - Recurrent and metastatic medullary thyroid carcinoma (MTC) remains difficult to treat due to its limited responsiveness to chemotherapy, radiotherapy, and imaging. To investigate an alternative therapeutic approach, we examined the feasibility of targeting gene-directed enzyme/prodrug therapy delivered by adenoviral vectors to MTC. We previously described a modified human calcitonin (CT)/CT gene-related peptide promoter that produced increased expression while maintaining specificity for MTC cells. In this study, we introduced an additional level of specificity by using cell-specific splicing and examined whether the selectivity of the gene-directed enzyme/prodrug therapy for MTC was enhanced when both the promoter and splicing features were combined in a single transcription unit. Two replication-defective adenoviruses were constructed that expressed the Escherichia coli purine nucleoside phosphorylase (PNP) gene under the transcriptional control of a modified T2 promoter (Ad.T2-PNP) or the T2 promoter in combination with a CT minigene cassette in which the PNP gene was imbedded within the CT gene exon 4 (Ad.T2-CT/PNP). The specificity of PNP expression by Ad.T2-PNP, Ad.T2-CT/PNP, and control viruses in the MTC cell line, TT, and in a panel of non-MTC cell lines was evaluated. The highest level of PNP gene expression and the most effective cell killing in the presence of prodrug occurred in TT cells infected with Ad.T2-PNP, followed by Ad.T2-CT/PNP. Infection of most non-MTC cell lines, even with high multiplicities of Ad.T2-PNP, produced only low-level PNP expression that resulted in minimal cell killing in the presence of prodrug. High-level expression of PNP and effective cell killing was observed with both adenoviral gene constructs. The highest level of cell specificity was achieved with the combined use of promoter and splicing regulation in the Ad.T2-CT/PNP virus. PMID- 12629125 TI - Influence of body surface area on serum peak thyrotropin (TSH) levels after recombinant human TSH administration. AB - Recombinant human TSH (rhTSH) has been proposed as an alternative method to the withdrawal of thyroid hormones in the follow-up of differentiated thyroid cancer. The aim of the present study was to evaluate the influence of several demographic and anthropometric parameters [age, body weight, height, body mass index, and body surface area (BSA)] on serum peak TSH levels after rhTSH administration. rhTSH was administered to 112 patients with differentiated thyroid carcinoma according to the conventional two-dose schedule (0.9 mg/d). Serum TSH levels were measured 24 h before and after the first administration of rhTSH, and then 24, 48, and 72 h after the second administration of rhTSH. In one severely obese patient, serum peak TSH values did not reach a valid stimulation range. Serum peak TSH levels were negatively related to body weight (r = -0.69; P < 0.0001), body mass index (r = -0.51; P < 0.0001), and BSA (r = -0.72; P < 0.0001). In a multivariate regression analysis including demographic and anthropometric variables, only BSA was independently associated to serum peak TSH concentrations (standardized beta coefficient = -0.721; P < 0.0001). In conclusion, body size seems to influence serum peak TSH levels after rhTSH administration. Future studies should evaluate the possibility of using personalized rhTSH doses, adjusted in relation to BSA. PMID- 12629126 TI - Metformin rapidly increases insulin receptor activation in human liver and signals preferentially through insulin-receptor substrate-2. AB - Metformin decreases endogenous glucose production by the liver. Few studies have examined the effect of metformin on the insulin-signaling pathway in liver models, and none have presented data on the effect in normal human liver. Huh7 human hepatoma cells and primary human hepatocytes were used. Insulin receptor (IR) and IR substrates (IRS)-1 and -2 were assessed by immunoprecipitation and immunoblot. Normal human liver was used to assay IR kinase activity (IR-KA). Tyrphostin AG1024 was used to inhibit IR-KA and examine effects on deoxyglucose uptake. Metformin (1 micro g/ml) increased IR tyrosine phosphorylation by 78% (P = 0.0007) in 30 min in human hepatocytes and Huh7 cells and increased IRS-2 but not IRS-1 activation, and the downstream increase in deoxyglucose uptake was mediated via increased translocation of GLUT-1 to the plasma membrane. Metformin did not augment maximal or submaximal insulin-stimulated IR activation. Metformin increased basal IR-KA by 150% (P = 0.0001). AG1024 inhibited metformin-induced IR beta phosphorylation in a concentration-dependent manner and abolished metformin induced 2-deoxyglucose uptake. This study demonstrates that the mechanism of action of metformin in liver involves IR activation, followed by selective IRS-2 activation, and increased glucose uptake via increased GLUT-1 translocation. The effect of metformin was completely blocked by an IR inhibitor. PMID- 12629127 TI - Zone-dependent expression of estrogen receptors alpha and beta in human benign prostatic hyperplasia. AB - Estrogen, which acts through estrogen receptors (ERs) alpha and beta, has been implicated in the pathogenesis of benign and malignant human prostatic tumors, i.e. benign prostatic hyperplasia and prostate cancer, thought to originate from different zones of the prostate [the transition zone (TZ) and peripheral zone (PZ), respectively]. Here, we examined the cellular distribution of ERalpha and ERbeta in human normal and hyperplastic prostate tissues, using in situ hybridization and immunohistochemistry. ERalpha expression was restricted to stromal cells of PZ. In contrast, ERbeta was expressed in the stromal cells of PZ as well as TZ. ERbeta-positive epithelial cells were evenly distributed in PZ and TZ of the prostate. Our results suggest that estrogen may play a crucial role in the pathogenesis of benign prostatic hyperplasia through ERbeta. PMID- 12629128 TI - Identification of a novel missense mutation that is as damaging to DAX-1 repressor function as a nonsense mutation. AB - Mutations in the DAX-1 (NROB1) gene result in X-linked congenital adrenal hypoplasia (AHC) and hypogonadotropic hypogonadism. The clinical presentation is usually as adrenal insufficiency in early life, with hypogonadotropic hypogonadism detected at the time of expected puberty. In this study we identified mutations in the DAX-1 gene of two patients with AHC. One mutation, Y399X, resulted in a premature stop codon and was associated with loss of Leydig cell responsiveness to human chorionic gonadotropin. The second, L297P, was a missense mutation, and human chorionic gonadotropin responsiveness was maintained. Kindred analysis established that the mutations had been inherited from the proband's mothers. The L297P has not been described previously and occurs within a highly conserved binding motif (LLXLXL). Transient transfection assays demonstrated that both mutations resulted in a severe loss of DAX-1 repressor activity. Immunohistochemical analysis of testicular tissue obtained from an affected sibling of the subject with the Y399X mutation, who had died with adrenal failure as a neonate, showed normal testicular morphology and expression of DAX-1, steroidogenic factor-1, and anti-Mullerian hormone protein. These data extend the clinical and molecular information on DAX-1 mutations, confirm normal testicular development at the neonatal stage, and illustrate variability in Leydig cell function. PMID- 12629129 TI - Differential expression, regulation, and induction of Smads, transforming growth factor-beta signal transduction pathway in leiomyoma, and myometrial smooth muscle cells and alteration by gonadotropin-releasing hormone analog. AB - The objective of this study was to further elucidate the role of TGFbeta and GnRH analog (GnRHa) in leiomyoma growth and regression. We examined the expression of Smads, TGFbeta receptor intracellular signaling molecules, in leiomyoma and myometrial smooth muscle cells (LSMC and MSMC), and determined whether TGFbeta and GnRHa differentially regulate their expression and induction in these cells. Using semiquantitative RT-PCR, Western blot analysis, and immunohistochemistry, we demonstrated that leiomyoma, myometrium, LSMC, and MSMC express receptor activated Smad3, common Smad4, and the inhibitory Smad7 mRNA and protein and showed that TGFbeta1, in a time-dependent manner, transiently induced Smad7 expression, with Smad3 and Smad4 remaining largely unchanged. TGFbeta1 increased the rate of Smad and phosphorylated Smad3 (pSmad3) induction in both cell types. Pretreatment with TGFbeta type II receptor antisense oligonucleotide resulted in a trend toward a lower TGFbeta-induced pSmad3. GnRHa, in a dose- and time dependent manner, increased the expression of Smad7 mRNA and the rapid induction of Smad3, Smad4, and Smad7 as well as pSmad3, which declined to control values at doses above 1 micro M in MSMC, but not in LSMC. GnRHa-induced pSamd3 was partly inhibited by a GnRH antagonist (antide). We concluded that leiomyoma, myometrium, LSMC, and MSMC express Smads, which are differentially expressed, induced, and activated by TGFbeta and are altered as a result of GnRHa treatment. These results suggest that TGFbeta and GnRHa mediate their actions through cross-talk involving Smads and most likely other signaling pathways that result in leiomyoma growth and regression. PMID- 12629130 TI - Antisense hTERT inhibits thyroid cancer cell growth. AB - Activation of telomerase represents an early step in carcinogenesis. Increased telomerase expression in malignant thyroid tumors suggests that inactivation of telomerase may represent a potential chemotherapeutic target. The purpose of this study was to inhibit the protein component of telomerase, hTERT, in a human thyroid cancer cell line in vitro and in vivo using an antisense strategy. A 235 bp fragment of hTERT cDNA was subcloned, and sense and antisense hTERT expression vectors were constructed. These vectors were transfected into a human thyroid carcinoma cell line (FRO). Tumorigenic potential was determined by cellular growth assay, rate of apoptosis, anchorage-independent growth, and tumor growth in a nude mouse model. Significant down-regulation of hTERT expression was seen in the antisense transfected cells, compared with control and those transfected with the sense vector. A decrease in telomerase activity by TRAP assay was observed in the antisense hTERT cells but not in cells transfected with the sense hTERT construct. Inhibition of cell growth was observed after approximately 20 population doublings in the antisense-hTERT clones and was associated with an increase in the rate of apoptosis and a change in cellular morphology. Moreover, anchorage-independent growth was reduced in vitro, and tumor growth rate was diminished in vivo in the antisense hTERT clones. Inhibition of telomerase activity with antisense hTERT in human thyroid cancer cells is achievable and may represent a novel target to inhibit tumor growth. PMID- 12629131 TI - Frequent methylation-associated silencing of the tissue inhibitor of metalloproteinase-3 gene in pancreatic endocrine tumors. AB - Molecular mechanisms contributing to the tumorigenesis of pancreatic endocrine tumors (PETs) are still not well understood. Allelic deletions at chromosome 22q12.3 were detected in about 30-60% of PETs, suggesting that inactivation of one or more tumor suppressor genes on this chromosomal arm is important for their pathogenesis. Because the putative tumor suppressor gene tissue inhibitor of metalloproteinase-3 (TIMP-3) has been located at 22q12.3, we undertook a genetic analysis of TIMP-3 to determine its role in the tumorigenesis of PETs. Single strand conformational polymorphism analysis, methylation-specific PCR, RNA expression analysis, and immunohistochemistry of TIMP-3 were performed in 21 sporadic PETs. Thirteen of 21 PETs (62%) revealed TIMP-3 alterations, including promoter hypermethylation and homozygous deletion. The predominant TIMP-3 alteration was promoter hypermethylation, identified in 8 of 18 (44%) PETs. It was tumor-specific and corresponded to loss or strong reduction of TIMP-3 protein expression. Notably, 11 of 14 (79%) PETs with metastases had TIMP-3 alterations, compared with only 1 of 7 (14%) PETs without metastases (P < 0.02). These data suggest a possibly important role of TIMP-3 in the tumorigenesis of human PETs, especially in the development of metastases, which has to be further evaluated in large-scale studies. PMID- 12629132 TI - Abnormally high expression of proteasome activator-gamma in thyroid neoplasm. AB - PA28-gamma is the activator of 20S proteasome, the ATP-dependent proteolytic system that plays an important role in cell cycle progression in various cell types. In this paper, we show the abnormally high expression of PA28-gamma in various thyroid neoplasms. Thyroid samples were obtained from patients with normal thyroid (4 cases) and with the following diseases: papillary adenocarcinoma (13 cases), multinodular goiter (4 cases), and anaplastic carcinoma (1 case). PA28-gamma expression was estimated by immunohistochemical staining and Western blotting. In all of the papillary adenocarcinoma samples, PA28-gamma was abnormally overexpressed, especially in cancer cells existing at the peripheral region of the cancer mass or in cancer cells invading the capsular region surrounding the cancer mass. In cancer cells of these areas, PA28-gamma was predominantly distributed in nucleus rather than in the cytoplasm of cancer cells. On the other hand, no obvious PA28-gamma expression was observed in the adjacent normal thyroid follicular cells. In multinodular goiter, the expression of PA28-gamma was relatively low compared with papillary adenocarcinoma. In anaplastic carcinoma, PA28-gamma was expressed at the highest level, especially in poorly differentiated regions such as squamous metaplasia of anaplastic cancer tissue. Therefore, the PA28-gamma expression seems to be restricted to thyroid cancer cells, especially in the region where the growth rate of cancer cells is accelerated. This result is further confirmed by the fact that C2, alpha-subunit of 20S proteasome, and proliferating cell nuclear antigen are similarly overexpressed in this region. Thus, PA28-gamma might be involved in the regulatory system for the cell cycle. Moreover, the growth of thyroid cancer cell lines was affected by the proteasome inhibitor, clasto-lactacystin beta-lactone. These results demonstrate that PA28-gamma is overexpressed in thyroid cancer, especially in its growth-accelerated cells. PMID- 12629133 TI - Type 3 iodothyronine deiodinase is highly expressed in the human uteroplacental unit and in fetal epithelium. AB - Type 3 iodothyronine deiodinase (D3) is the major physiologic inactivator of thyroid hormone. This selenoenzyme, previously identified in human placenta and brain, catalyzes the inner-ring deiodination of T(4) to reverse T(3) and T(3) to 3, 3'-diiodothyronine, both of which are biologically inactive. We analyzed D3 expression in several human adult and fetal tissues by immunohistochemistry and correlated the results with D3 activity assays where possible. High D3 expression was present in the placental syncytiotrophoblasts and cytotrophoblasts, endothelium of fetal vessels, and maternal decidua. D3 was also present at other sites of maternal-fetal interface, including the umbilical arteries and vein and the fetal respiratory, digestive, and urinary tract epithelium. Surprisingly, D3 was also present in the endometrial glands of nonpregnant human uteri, and endometrial activity approximated that of term placenta. The presence of D3 at maternal-fetal interfaces is consistent with its role in modulating the thyroid status of the human fetus and its expression in endometrium suggests that local regulation of thyroid status is important in implantation. PMID- 12629134 TI - Relationship between the growth hormone/insulin-like growth factor-I axis, insulin sensitivity, and adrenal androgens in normal prepubertal and pubertal girls. AB - The aim of this study was to analyze the possible implication of changes in the GH/IGF-I axis and in insulin sensitivity for the regulation of adrenal androgen secretion of normal prepubertal and adolescent girls. A total of 61 normal girls were evaluated in prepuberty [Group (Gr)1, n = 33; early (Gr1A, n = 16) and late (Gr1B, n = 17)]; puberty (Gr3, n = 28), early (Gr3A, n = 9) and late (Gr3B, n = 19); and during the transition between prepuberty and puberty (Gr2, n = 26). Insulin sensitivity was estimated by the fasting glucose/insulin ratio (G/I). In Gr1, G/I was significantly higher, and the mean serum IGF-I and serum dehydroepiandrosterone sulfate (DHEAS) were significantly lower than in Gr3 (P < 0.0001). Mean G/I in Gr1A and Gr3A was significantly higher than in Gr1B (P < 0.01) and Gr3B (P < 0.02), respectively, and ratios in Gr1B were also significantly higher than in Gr3A (P < 0.02). However, body mass index (BMI) in Gr1A, Gr1B, and Gr3A was not significantly different, although a significant increment was observed between late prepuberty (Gr1B) and late puberty (Gr3B; P < 0.0001). On the other hand, serum IGF-I levels in Gr1A and Gr3A were significantly lower than those in Gr1B (P < 0.01) and Gr3B (P < 0.02), respectively. The mean serum DHEAS level in Gr1A and Gr3A was significantly lower than in Gr1B (P < 0.01) and Gr3B (P < 0.02), respectively, and the level in Gr1B was also significantly lower than in Gr3A (P < 0.02). Correlation studies within Gr1, Gr2, and Gr3 were also performed. There was a significant positive correlation between serum DHEAS and age and a significant negative correlation between serum DHEAS and G/I in the three groups. However, a significant positive correlation between serum DHEAS and serum IGF-I was only found in Gr1. Furthermore, a significant negative correlation between BMI and the G/I was found in Gr2 and Gr3. Therefore, changes in insulin sensitivity might be involved in adrenal androgen synthesis both in prepuberty and in puberty, as well as during the transition from prepuberty to puberty. Changes in BMI suggest that adiposity might be a mediator of this effect, particularly during late puberty. On the other hand, the GH/IGF axis might be an important metabolic signal involved in the maturational changes of human adrenal androgens during prepuberty, at the time of adrenarche. Indeed, a significant negative correlation between G/I and serum IGF-I was found in Gr1, as well as in Gr2. In conclusion, the findings of this study indicate that the GH/IGF-I axis and insulin resistance might be involved in the mechanism of adrenarche during prepuberty in normal girls. Because these relationships had not been seen in boys, we proposed that prepubertal ovarian estrogens might be responsible for the sex difference. The relationship between insulin resistance and adrenal androgens persists during the transition from prepuberty to puberty, as well as during puberty. PMID- 12629135 TI - Resistin is expressed in the human placenta. AB - The mechanism for decreased insulin sensitivity in pregnant women is not fully clarified. Resistin, a novel peptide hormone, is specifically expressed in the adipose tissue and decreases insulin sensitivity in rodents. In the present study, we demonstrate resistin gene expression in the human placental tissue, mainly in trophoblastic cells. The resistin gene expression in term placental tissue was more prominent than was seen in the first trimester chorionic tissue. In contrast resistin gene expression in adipose tissue was rather weak and remained unchanged by pregnancy. Thus, resistin is a newly isolated placental hormone in humans which may modulate insulin sensitivity during pregnancy. PMID- 12629136 TI - UROMODULIN mutations cause familial juvenile hyperuricemic nephropathy. AB - Gout, which is commonly associated with hyperuricemia, affects 0.2% of the population. Hyperuricemia has a heterogeneous etiology that may be due to either over production and/or reduced renal clearance, of urate. In order to identify the mechanisms underlying reduced excretion of urate, we undertook positional cloning studies of familial juvenile hyperuricaemic nephropathy (FJHN), which is an autosomal dominant disorder characterized by hyperuricaemia, a low fractional renal excretion of urate, and chronic renal failure that is associated with interstitial fibrosis. The FJHN locus has been previously localized to a 22 centiMorgan interval flanked centromerically by D16S401 and telomerically by D16S3069, on chromosome 16p11-p13. This interval contains over 120 genes and we selected 13 renal expressed sequences to search for mutations in 5 unrelated FJHN families that contained 21 affected and 24 unaffected members. This revealed 5 heterozygous missense mutations (Cys77Tyr, Cys126Arg, Asn128Ser, Cys255Tyr and Cys300Gly) that altered evolutionary conserved residues in the gene encoding UROMODULIN. UROMODULIN, which is an 85 Kda glycoprotein, has roles in renal stone formation, the modulation of immune responses, and urothelial cytoprotection. The results of our studies, which have identified the gene causing FJHN, now indicate a further, novel role for UROMODULIN in urate metabolism. PMID- 12629137 TI - Efficacy of teriparatide and alendronate on nonvertebral fractures. PMID- 12629138 TI - Hyperglycemia: an independent marker of in-hospital mortality in patients with undiagnosed diabetes. PMID- 12629141 TI - Low levels of endogenous androgens increase the risk of atherosclerosis in elderly men: further supportive data. PMID- 12629143 TI - Recommended testing in patients with low bone density. PMID- 12629145 TI - Accessibility control and machinery of immunoglobulin class switch recombination. AB - Immunoglobulin (Ig) class switching is a process by which B lymphocytes shift from production of IgM to other Ig classes and subclasses via Ig class switch recombination (CSR). Multiple cellular and molecular processes are involved in CSR. Induction of a given IgH germline transcription initiates CSR processes. Ig germline transcription is selectively activated and induced by specific cytokine(s) via cytokine-specific signal pathways, synergized by CD40 signaling, and optimized by the 3' Ig alpha enhancers through locus control region function. Following Ig germline transcription, the switch-region DNA undergoes conformational changes so that it can serve as an appropriate substrate for nicking and cleavage by switch recombination machinery. Finally, the double strand breaks in donor and acceptor switch DNAs are processed, repaired, and ligated through a general nonhomologous end join pathway. CSR generates a new transcriptional unit for production of a class-switched Ig isotype. PMID- 12629146 TI - Regulation of lymphocyte-mediated killing by GTP-binding proteins. AB - Exocytosis of granules containing apoptosis-inducing proteins is one mechanism of target cell killing by cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells. Granules containing perforin and granzymes are redistributed to the area of cell contact initiated by specific interactions between surface ligands on a target cell and receptors on an effector lymphocyte. The formation of a stable conjugate between a cytotoxic lymphocyte and its potential target cell, followed by the directed delivery of granule components to the target cell are prerequisites of lymphocyte-mediated killing. Critical to understanding the development of cytotoxic function by CTLs and NK cells is the delineation of the second messenger pathways that specifically control the reorganization of the actin cytoskeleton during cell-mediated cytotoxicity. The low molecular weight guanosine 5'-triphosphate-binding proteins of the Rho family play a central role in these regulatory events controlling cytotoxic lymphocyte activation. PMID- 12629147 TI - Alerting and tuning the immune response by extracellular nucleotides. AB - The interplay between pro- and anti-inflammatory mechanisms during inflammatory and immune responses is critical for avoiding excessive tissue damage. Extracellular nucleotides (e.g., adenosine 5'-triphosphate) may represent constitutive signals that can alert the immune system of abnormal cell death. Relatively high doses of nucleotides induce rapid release of proinflammatory mediators and favor pathogen killing. However, recent findings on antigen presenting cells, particularly dendritic cells, revealed a more complex role for these molecules. Chronic exposure to low-dose nucleotides can redirect cellular responses to prototypic activation stimuli, leading to suppressed inflammation and immune deviation. PMID- 12629148 TI - Inhibition of actin polymerization by peroxynitrite modulates neutrophil functional responses. AB - Peroxynitrite, a potent oxidant generated in inflammatory tissues, can nitrate tyrosine residues on a variety of proteins. Based on previous studies suggesting that actin might be a potential target for peroxynitrite-mediated nitration in neutrophils, we investigated the effects of peroxynitrite on actin function. We show here that peroxynitrite and the peroxynitrite generator (SIN-1) modified actin in a concentration-dependent manner, resulting in an inhibition of globular actin polymerization and filamentous-actin depolymerization in vitro. The effects of peroxynitrite were inhibited by the pyrrolopyrimidine antioxidant PNU-101033E, which has been shown previously to specifically block peroxynitrite-mediated tyrosine nitration. Furthermore, spectrophotometric and immunoblot analysis of peroxynitrite-treated actin demonstrated a concentration-dependent increase in nitrotyrosine, which was also blocked by PNU-101033E. Activation of neutrophils in the presence of a nitric oxide donor (S-nitroso-N-acetylpenicillamine) resulted in nitration of exogenously added actin. Nitrated actin was also found in peroxynitrite-treated neutrophils, suggesting that actin may be an important intracellular target during inflammation. To investigate this issue, we analyzed the effect of peroxynitrite treatment on a number of actin-dependent neutrophil processes. Indeed, neutrophil actin polymerization, migration, phagocytosis, and respiratory burst activity were all inhibited by SIN-1 treatment in a concentration-dependent manner. Therefore, the ability of peroxynitrite to inhibit actin dynamics has a significant effect on actin-dependent, cellular processes in phagocytic cells and may modulate their host defense function. PMID- 12629149 TI - The role of CCL22 (MDC) for the recruitment of eosinophils during allergic pleurisy in mice. AB - Eosinophils are important inflammatory cells in allergic diseases. In the present study, we have investigated the effects of CCL22 on the recruitment of eosinophils in vivo and in vitro. CCL22 induced a dose- and time-dependent recruitment of eosinophils into the pleural cavity of mice, and this was dependent on the release of platelet-activating factor (PAF) and subsequent generation of CCL11. However, in an allergic pleurisy model, an anti-CCL22 polyclonal antibody given during sensitization or before challenge had no significant effect on eosinophil recruitment. CCL22 did not induce eosinophil chemotaxis in vitro but was able to induce eosinophil degranulation in vitro and in vivo. In conclusion, we show that although exogenously added CCL22 may induce eosinophil migration in vivo via release of PAF and CCL11 (eotaxin), endogenous production of CCL22 does not drive eosinophil migration during allergic inflammation. However, CCL22 may be an important activator of eosinophils once these cells have migrated into tissue. PMID- 12629150 TI - Cross-talk between tumor cells and neutrophils through the Fas (APO-1, CD95)/FasL system: human glioma cells enhance cell viability and stimulate cytokine production in neutrophils. AB - Many tumor cells are resistant to Fas-mediated killing, which has been primarily used as a mechanism to evade immune attack. In this study, we found a new action of Fas on tumors where activation of the Fas signal may force tumor cells to produce survival factors for neutrophils. Human peripheral circulating neutrophils in coculture with glioma cells showed significant delays in spontaneous apoptosis. Interleukin (IL)-6 and IL-8 partially mediated the glioma cell-associated, protective effect on neutrophils. The Fas agonistic antibody CH 11 dose-dependently stimulated the expression of IL-6 and IL-8 in glioma cells. Accordingly, blocking the Fas/FasL interaction reduced IL-6 and IL-8 production in glioma cells and impaired their protective effect on neutrophils. Coculture with glioma cells also affected the expression of cytokines in neutrophils, including IL-8, interferon-gamma, and tumor necrosis factor alpha to various extents. Collectively, our results demonstrate bi-directional cross-talk between tumor and immune cells. Although Fas activation alone cannot induce apoptosis in tumor cells, it may potentially initiate an effective anti-tumor response through a circumvented mechanism. PMID- 12629151 TI - Toxicity of human monocytic THP-1 cells and microglia toward SH-SY5Y neuroblastoma cells is reduced by inhibitors of 5-lipoxygenase and its activating protein FLAP. AB - To explore whether the proinflammatory products of the 5-lipoxygenase (5-LOX) pathway are involved in microglia-mediated toxicity toward neuronal cells, we evaluated the effects of 5-LOX inhibitors using an in vitro assay system where human neuronal SH-SY5Y cells are exposed to toxic secretions from THP-1 monocytic cells or human microglia. The specific 5-LOX inhibitors, REV 5901, zileuton, and 5-hydroxyeicosatetraenoic acid lactone; the nonselective LOX inhibitors, phenidone and dapsone; the dual 5-LOX/cyclooxygenase inhibitor, tepoxalin; and the selective inhibitor of the 5-LOX-activating protein (FLAP), MK-886, inhibited such toxicity. The toxicity was enhanced by the 5-LOX product leukotriene (LT)D(4) and reduced by the selective cysteinyl LT receptor (CysLT(1)) antagonist MK-571. The mRNAs for 5-LOX and FLAP were detected in THP-1 cells and human microglia but not in SH-SY5Y cells. The data suggest that inhibition of proinflammatory LT production by 5-LOX inhibition could selectively reduce toxicity of microglial cells and thus be beneficial in neuroinflammatory diseases. PMID- 12629152 TI - Differentiation of stress, metabolism, communication, and defense responses following transplantation. AB - The biological complexity of allograft rejection and alloantigen-independent mechanisms is poorly understood. Therefore, we analyzed four components of the biological response following transplantation by global gene analysis. A comparative and kinetic approach was used to identify gene expression profiles. Biological processes were assigned to genes displaying the largest alterations in expression. Metabolism, stress response, and cell organization were the predominant, biological processes associated with ischemia and systemic stress. Innate and adaptive immune responses induced a transcriptional shift toward defense and cell communication. The kinetic analysis showed a shift from innate toward adaptive responses in the post-transplant course. PMID- 12629153 TI - Beta 1/beta 3 integrin ligation is uncoupled from ERK1/ERK2 activation in cytotoxic T lymphocytes. AB - beta 3 integrins mediate fibronectin binding and enhanced activation of cytotoxic T lymphocytes (CTL). The intracellular signals initiated by beta 3 integrins in lymphocytes are not well characterized, but in many cell types, beta 1 integrin ligation activates mitogen-activated protein (MAP) kinases. In the present study, we find that fibronectin can synergize with very low levels of CD3 stimulation to activate the extracellular signal-regulated kinase (ERK)1 and ERK2 MAP kinases but that fibronectin alone induces no detectable MAP kinase activation in CTL. Surprisingly, antibodies to beta1 or beta 3 integrins were also unable to stimulate MAP kinase activation, suggesting that although beta 1 integrins are capable of stimulating MAP kinase activation in other cells, they cannot do so in CTL. In CTL, phosphorylation of proline-rich tyrosine kinase 2 downstream of integrin stimulation did not result in recruitment of the adaptor protein Grb2. Additionally, we examined the role of MAP kinases in regulating integrin-mediated adhesion. Anti-CD3-triggered adhesion to fibronectin was largely insensitive to the MAP kinase kinase inhibitor PD98059. Triggered cell-spreading on fibronectin was inhibited by PD98059 but not by U0126. In summary, ligation of beta 3 integrin by antibodies or fibronectin or of beta1 integrin by monoclonal antibodies fails to activate ERK MAP kinases, but integrin ligation synergizes with T cell receptor stimulation upstream of MAP kinases. PMID- 12629154 TI - Possible contribution of apoptosis-inducing factor (AIF) and reactive oxygen species (ROS) to UVB-induced caspase-independent cell death in the T cell line Jurkat. AB - We analyzed the mechanism of UVB-induced cell death using the Jurkat T cell line. Apoptosis was assessed by measuring phosphatidylserine (PS) externalization, caspase activity, the decrease in mitochondrial membrane potential (Delta Psi m), nucleosomal DNA fragmentation, and morphological changes such as chromatin condensation. The mitochondrio-nuclear translocation of apoptosis-inducing factor (AIF) was evaluated by confocal laser microscopy. The cell death pattern of UVB irradiated cells was similar to the Fas-induced cell death pattern. However, zVAD fmk inhibited the nucleosomal fragmentation of DNA but not the externalization of PS, decrease in Delta Psi m, or mitochondrio-nuclear translocation of AIF. N acetyl L-cysteine significantly inhibited the translocation of AIF induced by UVB. These results suggested that caspase-dependent and -independent pathways were involved in UVB-induced cell death in Jurkat cells, and the mitochondrio nuclear translocation of AIF was associated with the latter pathway. In addition, reactive oxygen species generated by UVB might be involved in inducing the mitochondrio-nuclear translocation of AIF. PMID- 12629155 TI - Human immunodeficiency virus type 1 (HIV-1) induces activation of multiple STATs in CD4+ cells of lymphocyte or monocyte/macrophage lineages. AB - Human immunodeficiency virus type 1 (HIV-1) impacts the activation state of multiple lineages of hematopoietic cells. Chronic HIV-1 infection among individuals with progressive disease can be associated with increased levels of activated signal transducers and activators of transcription (STATs) in peripheral blood mononuclear cells. To investigate interactions between HIV-1 and CD4(+) cells, activated, phosphorylated STAT proteins in nuclear extracts from lymphocytic and promonocytic cell lines as well as primary monocyte-derived macrophages were measured. Levels of activated STATs increased six- to tenfold in HUT78 and U937 cells within 2 h following exposure to virions. The response to virus was dose-dependent, but kinetics of activation was delayed relative to interleukin-2 or interferon-gamma. Activation of STAT1, STAT3, and STAT5 occurred with diverse viral envelope proteins, independent of coreceptor use or viral replication. Envelope-deficient virions had no effect on STAT activation. Monoclonal antibody engagement of CD4 identified a novel role for CD4 as a mediator in the activation of multiple STATs. Results provide a model for HIV-1 pathogenesis in infected and noninfected hematopoietic cells. PMID- 12629156 TI - The wake-promoting hypocretin-orexin neurons are in an intrinsic state of membrane depolarization. AB - Wakefulness depends on the activity of hypocretin-orexin neurons because their lesion results in narcolepsy. How these neurons maintain their activity to promote wakefulness is not known. Here, by recording for the first time from hypocretin-orexin neurons and comparing their properties with those of neurons expressing melanin-concentrating hormone, we show that hypocretin-orexin neurons are in an intrinsic state of membrane depolarization that promotes their spontaneous activity. We propose that wakefulness and associated energy expenditure thus depend on that property, which allows the hypocretin-orexin neurons to maintain a tonic excitatory influence on the central arousal and peripheral sympathetic systems. PMID- 12629157 TI - Plasma membrane GABA transporters reside on distinct vesicles and undergo rapid regulated recycling. AB - Plasma membrane neurotransmitter transporters affect synaptic signaling through transmitter sequestration. Transporters redistribute to and from the plasma membrane, suggesting a role for trafficking in regulating synaptic transmitter levels. One method for controlling transmitter levels would be to regulate transporter redistribution in parallel with transmitter release. Thus, how similar are these processes? We show that the trafficking of the GABA transporter GAT1 resembles the trafficking of neurotransmitter-filled synaptic vesicles: (1) transporters located on the plasma membrane are internalized and reinserted into the plasma membrane on the order of minutes; (2) the rate of recycling is depolarization and calcium dependent; (3) GAT1 internalization is associated with clathrin and dynamin; and (4) intracellular GAT1 is associated with multiple compartments and, more importantly, is found on a distinct class of vesicles. These vesicles are clear, approximately 50 nm in diameter, and contain many proteins found on neurotransmitter-containing small synaptic vesicles; however, they appear to lack several traditional small synaptic vesicle proteins, such as synaptophysin and the vesicular GABA transporter. These data provide additional support for the hypothesis that GABA transporters traffic in parallel with neurotransmitter-containing small synaptic vesicles and also raise the possibility that some fraction of vesicles found in GABAergic neurons may not be participating in transmitter release but rather in the rapid regulated redistribution of membrane proteins involved in transmitter uptake. PMID- 12629158 TI - Absence of Ndn, encoding the Prader-Willi syndrome-deleted gene necdin, results in congenital deficiency of central respiratory drive in neonatal mice. AB - necdin (Ndn) is one of a cluster of genes deleted in the neurodevelopmental disorder Prader-Willi syndrome. necdin is upregulated during neuronal differentiation and is thought to play a role in cell cycle arrest in terminally differentiated neurons. Most necdin-deficient Ndn(tm2Stw) mutant pups carrying a targeted replacement of Ndn with a lacZ reporter gene die in the neonatal period of apparent respiratory insufficiency. We now demonstrate that the defect can be explained by abnormal neuronal activity within the putative respiratory rhythm generating center, the pre-Botzinger complex. Specifically, the rhythm is unstable with prolonged periods of depression of respiratory rhythmogenesis. These observations suggest that the developing respiratory center is particularly sensitive to loss of necdin activity and may reflect abnormalities of respiratory rhythm-generating neurons or conditioning neuromodulatory drive. We propose that necdin deficiency may contribute to observed respiratory abnormalities in individuals with Prader-Willi syndrome through a similar suppression of central respiratory drive. PMID- 12629159 TI - Identification of calcineurin as a key signal in the extinction of fear memory. AB - Memory extinction refers to a gradual decrease of the previously acquired response when exposed to conditional stimulus without pairing with unconditional stimulus. Here we show for the first time that fear training-induced phosphorylation of specific substrates in the rat amygdala is reduced after extinction trials and is accompanied by an increase in the protein level and enzymatic activity of calcineurin. In parallel, calcineurin inhibitors prevented extinction-induced protein dephosphorylation as well as extinction of fear memory. Thus, extinction training increased phosphatase activity likely via an expression of calcineurin. Calcineurin then created a negative-feedback loop and directly or indirectly dephosphorylated specific substrates, which, in their phosphorylated state, were required for memory consolidation. Accordingly, in our experimental condition, extinction could be ascribed at least in part to a weakening of the original signaling. PMID- 12629160 TI - Calcium-dependent exocytosis of atrial natriuretic peptide from astrocytes. AB - Astrocytes are non-neuronal cells in the CNS, which, like neurons, are capable of releasing neuroactive molecules. However, the mechanism of release is ill defined. In this study, we investigated the mechanism of release of atrial natriuretic peptide (ANP) from cultured cortical astrocytes by confocal microscopy. To study the discharge of this hormone, we transfected astrocytes with a construct to express pro-ANP fused with the emerald green fluorescent protein (ANP.emd). The transfection of cells with ANP.emd resulted in fluorescent puncta in the cytoplasm that represent secretory organelles. If ANP is released by exocytosis, in which the vesicle fuses with the plasma membrane, then the total intensity of the green fluorescing probe should decrease, whereas the vesicle membrane is incorporated into the plasma membrane. To monitor exocytosis, we labeled the membrane with the fluorescent styryldye FM 4-64, a reporter of cumulative exocytosis. The application of ionomycin to elevate cytoplasmic [Ca(2+)] increased the fluorescence intensity of FM 4-64, whereas that of ANP.emd decreased. These effects were not observed in the absence of extracellular Ca(2+), suggesting that ANP is released by regulated Ca(2+)-dependent exocytosis from astrocytes. PMID- 12629161 TI - Opposing roles of D1 and D2 receptors in appetitive conditioning. AB - Previous studies have shown that D(1) receptor blockade disrupts and D(2) receptor blockade enhances long-term potentiation. These data lead to the prediction that D(1) antagonists will attenuate and D(2) antagonists will potentiate at least some types of learning. The prediction is difficult to test, however, because disruptions in either D(1) or D(2) transmission lead to reduced locomotion, exploration, and response execution and are therefore likely to impair learning that requires behavioral responding (including exploration of an environment) during the learning episode. Under a paradigm that minimizes motor requirements, rats were trained to enter a food compartment during pellet presentation. Animals then received tone-food pairings under the influence of D(1) antagonist SCH23390 (0, 0.4, 0.8, and 0.16 mg/kg) or D(2) antagonist raclopride (0, 0.2, 0.4, and 0.8 mg/kg). An additional group received unpaired presentations of tone and food. On a drug-free test day 24 hr later, animals that had been under the influence of SCH23390 (like animals that had received unpaired presentations of tone and food) showed reduced head entries in response to the tone, whereas animals that had been under the influence of raclopride showed increased head entries in response to the tone compared with vehicle controls. These data demonstrate that, under a conditioned approach paradigm, D(1) and D(2) family receptor antagonists disrupt and promote learning, respectively, as predicted by the effects of D(1) and D(2) receptor blockade on neuronal plasticity. PMID- 12629162 TI - Gonadal hormones affect spine synaptic density in the CA1 hippocampal subfield of male rats. AB - The effects of androgen on the density of spine synapses on pyramidal neurons in the CA1 area of the hippocampus were studied in male rats. Gonadectomy (GDNX) had no significant effect on the number of CA1 pyramidal cells but reduced CA1 spine synapse density by almost 50% (to 0.468 +/- 0.018 spine synapses/microm(3)) compared with sham-operated controls (0.917 +/- 0.06 spine synapses/microm(3)). Treatment of GDNX rats with testosterone propionate (500 microg/d, s.c., 2 d) increased spine synapse density to levels (1.01 +/- 0.026 spine synapses/microm(3)) comparable with intact males. A similar increase in synapse density (1.013 +/- 0.05 spine synapses/microm(3)) was observed in GDNX animals after treatment with dihydrotestosterone (DHT) (500 microg/d, s.c., 2 d) but not after estradiol (10 microg/d, s.c., 2 d; 0.455 +/- 0.02 spine synapse/microm(3)). These data indicate that testosterone is important for maintenance of normal spine synapse density in the CA1 region of the male rat hippocampus. The comparable responses to testosterone and the non-aromatizable androgen DHT, coupled with the lack of response to estradiol, suggest that testosterone acts directly on hippocampal androgen receptors rather than indirectly via local estrogen biosynthesis. PMID- 12629163 TI - Afterhyperpolarization regulates firing rate in neurons of the suprachiasmatic nucleus. AB - Cluster I neurons of the suprachiasmatic nucleus (SCN), which are thought to be pacemakers supporting circadian activity, fire spontaneous action potentials that are followed by a monophasic afterhyperpolarization (AHP). Using a brain slice preparation, we have found that the AHP has a shorter duration in cells firing at higher frequency, consistent with circadian modulation of the AHP. The AHP is supported by at least three subtypes of K(Ca) channels, including apamin sensitive channels, iberiotoxin-sensitive channels, and channels that are insensitive to both of these antagonists. The latter K(Ca) channel subtype is involved in rate-dependent regulation of the AHP. Voltage-clamped, whole-cell Ca(2+) channel currents recorded from SCN neurons were dissected pharmacologically, revealing all of the major high-voltage activated subtypes: L , N-, P/Q-, and R-type Ca(2+) channel currents. Application of Ca(2+) channel antagonists to spontaneously firing neurons indicated that predominantly L- and R type currents trigger the AHP. Our findings suggest that apamin- and iberiotoxin insensitive K(Ca) channels are subject to diurnal modulation by the circadian clock and that this modulation either directly or indirectly leads to the expression of a circadian rhythm in spiking frequency. PMID- 12629164 TI - Interleukin-1 mediates pathological effects of microglia on tau phosphorylation and on synaptophysin synthesis in cortical neurons through a p38-MAPK pathway. AB - The presence of tangles of abnormally phosphorylated tau is a characteristic of Alzheimer's disease (AD), and the loss of synapses correlates with the degree of dementia. In addition, the overexpression of interleukin-1 (IL-1) has been implicated in tangle formation in AD. As a direct test of the requirement for IL 1 in tau phosphorylation and synaptophysin expression, IL-1 actions in neuron microglia cocultures were manipulated. Activation of microglia with secreted beta amyloid precursor protein or lipopolysaccharide elevated their expression of IL 1alpha, IL-1beta, and tumor necrosis factor alpha (TNFalpha) mRNA. When such activated microglia were placed in coculture with primary neocortical neurons, a significant increase in the phosphorylation of neuronal tau was accompanied by a decline in synaptophysin levels. Similar effects were evoked by treatment of neurons with recombinant IL-1beta. IL-1 receptor antagonist (IL-1ra) as well as anti-IL-1beta antibody attenuated the influence of activated microglia on neuronal tau and synaptophysin, but anti-TNFalpha antibody was ineffective. Some effects of microglial activation on neurons appear to be mediated by activation of p38 mitogen-activated protein kinase (p38-MAPK), because activated microglia stimulated p38-MAPK phosphorylation in neurons, and an inhibitor of p38-MAPK reversed the influence of IL-1beta on tau phosphorylation and synaptophysin levels. Our results, together with previous observations, suggest that activated microglia may contribute to neurofibrillary pathology in AD through their production of IL-1, activation of neuronal p38-MAPK, and resultant changes in neuronal cytoskeletal and synaptic elements. PMID- 12629165 TI - Calcium dynamics, buffering, and buffer saturation in the boutons of dentate granule-cell axons in the hilus. AB - The axons of dentate gyrus granule cells form synapses in the hilus. Ca(2+) signaling was investigated in the boutons of these axons using confocal fluorescence imaging. Boutons were loaded with various concentrations of the Ca(2+) indicator Oregon Green BAPTA-1 by patch-clamping the cell bodies and allowing the dye to diffuse into the axon. Resting free [Ca(2+)] started at 74 nm, rose to approximately 1 microm immediately after an action potential, and then decayed to rest with a time constant of 43 msec (all extrapolated to a dye concentration of zero). Action potential-induced [Ca(2+)] rises were smaller in larger boutons, consistent with a size-independent Ca(2+) channel density of 45/microm(2). Action potential-induced [Ca(2+)] changes varied with dye concentration in a manner consistent with kappa(E) approximately 20 for the ratio of endogenous buffer-bound Ca(2+) to free Ca(2+). During trains of action potentials, [Ca(2+)] increments summed supralinearly by more than that expected from dye saturation. The amount of endogenous Ca(2+) buffering declined as [Ca(2+)] rose, and this saturation indicated a buffer with a dissociation constant of approximately 500 nm and a concentration of approximately 130 microm. This is similar to the dissociation constant of calbindin-D28K, a Ca(2+)-binding protein that is abundant in dentate granule cells. Thus, calbindin-D28K is a good candidate for the Ca(2+) buffer revealed by these experiments. The saturation of endogenous buffer can generate short-term facilitation by amplifying [Ca(2+)] changes during repetitive activity. Buffer saturation may also be relevant to the presynaptic induction of long-term potentiation at synapses formed by dentate granule cells. PMID- 12629166 TI - Dopamine modulation of perisomatic and peridendritic inhibition in prefrontal cortex. AB - The computations underlying cognitive functions are performed by a diversity of interactions between interneurons and pyramidal neurons that are subject to modulatory influences. Here we have used paired whole-cell recording to study the influence of dopamine on local inhibitory circuits involving fast-spiking (FS) and non-FS cells, respectively. We found that dopamine depressed inhibitory transmission between FS interneurons and pyramidal neurons but enhanced inhibition between non-FS interneurons and pyramidal cells. FS inhibitory transmission exhibited properties associated with presynaptic action at D(1) receptors that were not evident in non-FS inhibitory connections. In addition, FS and non-FS interneurons differed morphologically, forming contacts on the perisomatic and peridendritic domains, respectively, of their pyramidal cell targets. These findings provide evidence for both a dual mode of inhibition in prefrontal circuitry and circuit-dependent modulation by dopamine. PMID- 12629167 TI - A mutation in Af4 is predicted to cause cerebellar ataxia and cataracts in the robotic mouse. AB - The robotic mouse is an autosomal dominant mutant that arose from a large-scale chemical mutagenesis program. It has a jerky, ataxic gait and develops adult onset Purkinje cell loss in the cerebellum in a striking region-specific pattern, as well as cataracts. Genetic and physical mapping of the disease locus led to the identification of a missense mutation in a highly conserved region of Af4, a putative transcription factor that has been previously implicated in leukemogenesis. We demonstrate that Af4 is specifically expressed in Purkinje cells, and we hypothesize that the expression of mutant Af4 leads to neurodegeneration. This function was not identified through knock-out studies, highlighting the power of phenotype-driven mutagenesis in the mouse to identify new pathways involved in neurological disease. PMID- 12629168 TI - Role of integrin-linked kinase in nerve growth factor-stimulated neurite outgrowth. AB - The role of integrin-linked kinase (ILK), a kinase that is involved in various cellular processes, including adhesion and migration, has not been studied in primary neurons. Using mRNA dot blot and Western blot analysis of ILK in rat and human brain tissue, we found that ILK is expressed in various regions of the CNS. Immunohistochemical and immunocytochemical techniques revealed granular ILK staining that is enriched in neurons and colocalizes with the beta1 integrin subunit. The role of ILK in neurite growth promotion by NGF was studied in rat pheochromocytoma cells and dorsal root ganglion neurons using a pharmacological inhibitor of ILK (KP-392) or after overexpression of dominant-negative ILK (ILK DN). Both molecular and pharmacological inhibition of ILK activity significantly reduced NGF-induced neurite outgrowth. Survival assays indicate that KP-392 induced suppression of neurite outgrowth occurred in the absence of cell death. ILK kinase activity was stimulated by NGF. NGF-mediated stimulation of phosphorylation of both AKT and the Tau kinase glycogen synthase kinase-3 (GSK-3) was inhibited in the presence of KP-392 and after overexpression of ILK-DN. Consequently, ILK inhibition resulted in an increase in the hyperphosphorylation of Tau, a substrate of GSK-3. Together these findings indicate that ILK is an important effector in NGF-mediated neurite outgrowth. PMID- 12629169 TI - The E2F-Cdc2 cell-cycle pathway specifically mediates activity deprivation induced apoptosis of postmitotic neurons. AB - Neuronal apoptosis plays a critical role in the normal development of the mammalian brain and is thought to contribute to the pathogenesis of several neurologic disorders. However, the intracellular mechanisms underlying apoptosis of neurons remain incompletely understood. In the present study, we characterized a cell-cycle-based mechanism by which neuronal activity deprivation induces apoptosis of postmitotic neurons. Activity deprivation, but not growth factor withdrawal, was found to induce Cdc2 expression and consequent Cdc2-mediated apoptosis in granule neurons of the developing rat cerebellum. We found that activity deprivation induces cdc2 transcription in neurons via an E2F-binding element (EBE) within the cdc2 promoter. The transcription factor E2F1 that is expressed in granule neurons was found in DNA binding assays to bind to the EBE of the cdc2 gene. In chromatin immunoprecipitation analysis, endogenous E2F1 forms a complex with the promoter of the endogenous cdc2 gene in granule neurons, indicating that endogenous E2F1 is poised to activate transcription of the endogenous cdc2 gene in neurons. Consistent with this conclusion, a dominant interfering form of E2F, when expressed in granule neurons, blocked activity deprivation-induced cdc2 transcription. In other experiments, we found that the expression of E2F1 in granule neurons induces Cdc2 expression and promotes neuronal apoptosis via the activation of Cdc2. Remarkably, in contrast to inducing the E2F-mediated expression and activation of Cdc2 in granule neurons, activity deprivation fails to stimulate the expression of E2F-target genes that trigger DNA synthesis and replication. Together, our findings define a novel apoptotic mechanism whereby E2F selectively couples an activity deprivation induced signal to cdc2 transcription in the absence of stimulating DNA synthesis and thus culminating in Cdc2-mediated apoptosis of postmitotic neurons. PMID- 12629170 TI - Glial cell inhibition of neurons by release of ATP. AB - ATP is released by neurons and functions as a neurotransmitter and modulator in the CNS. Here I show that ATP released from glial cells can also serve as a potent neuromodulator, inhibiting neurons in the retina of the rat. Activation of glial cells by focal ejection of ATP, ATPgammaS, dopamine, thrombin, or lysophosphatidic acid or by mechanical stimulation evoked hyperpolarizing responses and outward currents in a subset of retinal ganglion cells by increasing a Ba(2+)-sensitive K(+) conductance in the neurons. This glia-evoked inhibition reduced the firing rate of those neurons that displayed spontaneous spike activity. The inhibition was abolished by the A(1) adenosine receptor antagonist DPCPX (8-cyclopentyl-1,3-dipropylxanthine) (10 nm) and was reduced by the ecto-ATPase inhibitor ARL-67156 (6-N,N-diethyl-D-beta,gamma dibromomethyleneATP) (50 microm) and by the ectonucleotidase inhibitor AOPCP [adenosine-5'-O-(alpha,beta-methylene)-diphosphonate] (250 microm). Selective activation of retinal glial cells demonstrated that Muller cells, but not astrocytes, mediate the inhibition. ATP release from Muller cells into the inner plexiform layer of the retina was shown using the luciferin-luciferase chemiluminescence assay. These findings demonstrate that activated glial cells can inhibit neurons in the retina by the release of ATP, which is converted to adenosine by ectoenzymes and subsequently activates neuronal adenosine receptors. The results lend support to the hypothesis that glial cells play an active role in information processing in the CNS. PMID- 12629171 TI - Interaction between liprin-alpha and GIT1 is required for AMPA receptor targeting. AB - Liprin-alpha is a multidomain protein that interacts with the LAR family of receptor protein tyrosine phosphatases and the GRIP/ABP family of AMPA receptor interacting proteins. Previous studies have indicated that liprin-alpha regulates the development of presynaptic active zones and that the association of liprin alpha with GRIP is required for postsynaptic targeting of AMPA receptors. However, the underlying molecular mechanisms are not well understood. Here we report that liprin-alpha directly interacts with GIT1, a multidomain protein with GTPase-activating protein activity for the ADP-ribosylation factor family of small GTPases known to regulate protein trafficking and the actin cytoskeleton. Electron microscopic analysis indicates that GIT1 distributes to the region of postsynaptic density (PSD) as well as presynaptic active zones. GIT1 is enriched in PSD fractions and forms a complex with liprin-alpha, GRIP, and AMPA receptors in brain. Expression of dominant-negative constructs interfering with the GIT1 liprin-alpha interaction leads to a selective and marked reduction in the dendritic and surface clustering of AMPA receptors in cultured neurons. These results suggest that the GIT1-liprin-alpha interaction is required for AMPA receptor targeting and that GIT1 may play an important role in the organization of presynaptic and postsynaptic multiprotein complexes. PMID- 12629173 TI - Impaired spinal cord glutamate transport capacity and reduced sensitivity to riluzole in a transgenic superoxide dismutase mutant rat model of amyotrophic lateral sclerosis. AB - We characterized synaptosomal glutamate transport activity in a recently developed transgenic rat model of amyotrophic lateral sclerosis (ALS) overexpressing the G93A Cu(2+)/Zn(2+) superoxide dismutase (SOD1) mutation. Using spinal cord synaptosomes, a significant reduction (43%) in the maximal velocity for high-affinity, Na(+)-dependent glutamate uptake was observed at disease end stage in G93A rats compared with age-matched controls. Similarly, a 27% reduction in maximum velocity (V(max)) was measured at disease onset, but no difference in spinal cord V(max) values were observed with presymptomatic animals compared with controls. In comparison, we observed no differences in the V(max) for glutamate clearance at disease end stage with synaptosomes from cortex, hippocampus, striatum, cerebellum, and brainstem, indicating a specific deficit in the spinal cord. The pharmacological sensitivity of spinal cord uptake to dihydrokainate suggests that the GLT-1 (glutamate transporter-1) subtype primarily mediates the transport activity. Expression analysis revealed a loss of GLT-1 as well as qualitative changes in GLAST (glutamate/aspartate transporter) but no measurable changes in EAAC1 (excitatory amino acid carrier 1) in spinal cord of end-stage G93A rats, indicating that deficits in glutamate transporters in this rat model may be glial specific. Riluzole, a neuroprotective agent used clinically to slow the progression of ALS, produced an enhancement of spinal cord synaptosomal glutamate uptake in control animals and early-stage disease G93A rats, but this effect was lost in end-stage animals. Altered expression of astroglial glutamate transporters accompanied by reduced capacity for spinal cord clearance of extracellular glutamate in the G93A SOD1 transgenic rat may account for a dampened effect of riluzole to enhance glutamate uptake at end-stage disease. PMID- 12629172 TI - Metabotropic glutamate receptor activation enhances the activities of two types of Ca2+-activated k+ channels in rat hippocampal astrocytes. AB - The influence of activation of glutamate receptor (GluR) on outward K(+) current in cultured neonate rat hippocampal astrocytes was investigated. Patch-clamp analysis of K(+) channel currents in cultured astrocytes identified the existence of 71 +/- 6 and 161 +/- 11 pS single-channel K(+) currents that were sensitive to changes in voltage and [Ca(2+)](i) and blocked by external TEA but not by charybdotoxin, iberiotoxin, apamin, or 4-aminopyridine. Reverse transcriptase (RT)-PCR and Northern blot analysis revealed transcripts of the Ca(2+)-activated K(+) channel (K(Ca)) beta(4)-subunit (beta4) (KCNMB4) in cultured astrocytes. Expression of the metabotropic glutamate receptor (mGluR) subtypes mGluR1 and mGluR5 and the ionotropic glutamate receptor (iGluR) subtypes iGluR1 and iGluR4 were detected by RT-PCR and immunofluorescence analysis in cultured astrocytes. The mGluR agonists L-glutamate and quisqualate increased the open state probability (NP(o)) of the 71 and 161 pS K(+) channel currents that were prevented by the mGluR receptor antagonists 1-aminoindan-1,5-dicarboxylic acid or L-(+)-2-amino-3-phosphonopropionic acid and not by the iGluR antagonists (+)-5 methyl-10,11-dihydro-5H-dibenzo [a,d] cyclohepten-5,10-imine maleate or CNQX. Activation of the two types of K(+) channel currents by mGluR agonists was attenuated by pertussis toxin and by inhibition of phospholipase C (PLC) or cytochrome P450 arachidonate epoxygenase. These results indicate that brain astrocytes contain the KCNMB4 transcript and express two novel types of K(Ca) channels that are gated by activation of a G-protein coupled metabotropic glutamate receptor functionally linked to PLC and cytochrome P450 arachidonate epoxygenase activity. PMID- 12629174 TI - A regulated interaction of syntaxin 1A with the antidepressant-sensitive norepinephrine transporter establishes catecholamine clearance capacity. AB - Norepinephrine (NE) transporters (NETs) terminate noradrenergic synaptic transmission and represent a major therapeutic target for antidepressant medications. NETs and related transporters are under intrinsic regulation by receptor and kinase-linked pathways, and clarification of these pathways may suggest candidates for the development of novel therapeutic approaches. Syntaxin 1A, a presynaptic soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) protein, interacts with NET and modulates NET intrinsic activity. NETs colocalize with and bind to syntaxin 1A in both native preparations and heterologous systems. Protein kinase C activation disrupts surface NET/syntaxin 1A interactions and downregulates NET activity in a syntaxin dependent manner. Syntaxin 1A binds the NH(2) terminal domain of NET, and a deletion of this domain both eliminates NET/syntaxin 1A associations and prevents phorbol ester-triggered NET downregulation. Whereas syntaxin 1A supports the surface trafficking of NET proteins, its direct interaction with NET limits transporter catalytic function. These two contradictory roles of syntaxin 1A on NET appear to be linked and reveal a dynamic cycle of interactions that allow for the coordinated control between NE release and reuptake. PMID- 12629175 TI - Overexpression of copper/zinc superoxide dismutase in transgenic mice protects against neuronal cell death after transient focal ischemia by blocking activation of the Bad cell death signaling pathway. AB - The Bad signaling pathway contributes to the regulation of apoptosis after a variety of cell death stimuli, and Bad plays a key role in determining cell death or survival. We have reported that overexpression of copper/zinc superoxide dismutase (SOD1) reduces apoptotic cell death after transient focal cerebral ischemia (tFCI). However, both the role of the Bad pathway after tFCI and the role of oxygen free radicals in the regulation of apoptosis remain unknown. To clarify these issues, we used an in vivo tFCI model of SOD1 transgenic mice and wild-type mice. Moreover, to examine the role of protein kinase A (PKA) in the Bad pathway after tFCI, we administered the PKA inhibitor, H89, into the mouse brain after tFCI. Immunohistochemistry and Western blot analysis showed that dephosphorylation and translocation of Bad were detected early after tFCI and that they were promoted by H89 treatment but prevented by SOD1. Coimmunoprecipitation revealed that the dimerization of Bad progressed with 14-3 3 (Bad/14-3-3) and with Bcl-x(L) (Bad/Bcl-x(L)) after tFCI. Moreover, Bad/14-3-3 was prevented by H89 treatment but promoted by SOD1. Bad/Bcl-x(L) was prevented by SOD1 but promoted by H89 treatment. A cell death assay revealed that apoptotic related DNA fragmentation was aggravated by H89 treatment but reduced by SOD1. These results suggest that the Bad pathway mediated by PKA is involved in apoptotic cell death after tFCI and that overexpression of SOD1 may attenuate this apoptotic cell death. PMID- 12629176 TI - Multiple dose-dependent effects of Lis1 on cerebral cortical development. AB - Humans with heterozygous inactivating mutations of the Lis1 gene display type I lissencephaly, a severe form of cortical dysplasia hypothesized to result from abnormal neuronal migration. Previously we reported the construction of an allelic series of the Lis1 gene in mice to analyze the effects of graded reduction of LIS1 protein on the pathogenesis of this disorder and demonstrated a cell autonomous defect in neuronal migration (Hirotsune et al., 1998). Here we report the systematic examination of the consequences of dosage reduction of LIS1 on neocortical development using wild-type, null heterozygous (45% LIS1 protein), and compound null/hypomorphic (35% LIS1 protein) mice. The development of the preplate, Cajal-Retzius cells, and the radial glial scaffold appeared unaffected by LIS1 levels. However, a dose-dependent morphologic change in disorganization of the subplate was noted. LIS1 dose-dependent defects in neuronal migration were found in vivo and in vitro. The position and number of mitotic cells in the ventricular zone were more abnormal as LIS1 levels decreased, suggesting defects in interkinetic nuclear migration and neuroblast proliferation. LIS1 dose dependent progressive thinning of the cortex and ventricular zone occurred by programmed cell death. Thus, in addition to its requirement for cell autonomous neuronal migration, LIS1 influences the generation and survival of cortical ventricular zone neuroblasts. These studies reveal the importance of LIS1 levels in orderly cerebral cortical morphogenesis and suggest new insights into the pathogenesis of type I lissencephaly. PMID- 12629177 TI - Glycoprotein 130 signaling regulates Notch1 expression and activation in the self renewal of mammalian forebrain neural stem cells. AB - Glycoprotein130 (gp130) and Notch signaling are thought to participate in neural stem cell (NSC) self-renewal. We asked whether gp130 regulates Notch activity in forebrain epidermal growth factor (EGF)-responsive NSCs. Disruption of Notch1 using antisense or a gamma-secretase inhibitor demonstrated a requirement for Notch1 in the maintenance and proliferation of NSCs. Ciliary neurotrophic factor (CNTF) activation of gp130 in NSCs rapidly increased Notch1 expression. NOTCH1 activation, indicated by tumor necrosis factor alpha-converting enzyme (TACE)- and presenilin-mediated processing, also increased. Infusion of EGF+CNTF into adult forebrain lateral ventricles increased periventricular NOTCH1 compared with EGF alone. Neither Hes1 (hairy and enhancer of split) nor Hes5 appeared to mediate gp130-enhanced NOTCH1 signaling that regulates NSC maintenance. This is the first example of a link between gp130 signaling and NOTCH1 in regulating NSC self-renewal. PMID- 12629178 TI - Hippocampal neurogenesis follows kainic acid-induced apoptosis in neonatal rats. AB - The effects of kainic acid (KA) on neurogenesis in the developing rat hippocampus were investigated. Neonatal [postnatal day (P) 7] rats received a single bilateral intracerebroventricular infusion of KA (50 nmol in 1.0 microl) or vehicle. At P14, P25, P40, and P60, the spatial and temporal relationships between the neurodegeneration and neurogenesis induced by KA were explored using terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL) to detect the dying cells and 5-bromodeoxyuridine (BrdU) to label newly generated cells. There was progressive loss of neurons in the cornu ammonis (CA) 1 and CA3 subfields of the hippocampus at all time points in KA-treated rats. TUNEL staining identified dying cells at P14 through P60, mainly in the CA3 subfield. The number of TUNEL-positive cells decreased with age. Neurogenesis also was observed in the KA-treated hippocampus. The number of BrdU-positive cells in the dentate gyrus was significantly decreased at P14, when the number of TUNEL-positive cells is highest. However, at later time points (P40 and P60) the number of BrdU-positive cells in the dentate gyrus was significantly increased. In addition, the number of BrdU-positive cells was increased in the CA3 subfield at P40 and P60 in KA-treated rats. A substantial proportion (40%) of the newly generated cells in CA3 also expressed markers of immature and mature neurons (class III beta-tubulin and neuronal nuclei). Newly generated cells in the CA3 subfield only rarely expressed glial markers (8%). These results suggest that a single exposure to KA at P7 has both immediate (inhibition) and delayed (stimulation) effects on neurogenesis within the dentate gyrus of developing rats. KA administration resulted in both neuronal apoptosis and neurogenesis within the CA3 subfield, suggesting that the purpose of neurogenesis in the CA3 is to replace neurons lost to apoptosis. PMID- 12629179 TI - Segregated expression of AMPA-type glutamate receptors and glutamate transporters defines distinct astrocyte populations in the mouse hippocampus. AB - Recent data have suggested the existence of direct signaling pathways between glial cells and neurons. Here we report the coexistence of distinct types of cells expressing astrocyte-specific markers within the hippocampus that display diverse morphological, molecular, and functional profiles. Usage of transgenic mice with GFAP promoter-controlled enhanced green fluorescent protein (EGFP) expression allowed the identification of astroglial cells after fresh isolation or in brain slices. Combining patch-clamp recordings and single-cell reverse transcription-PCR, we distinguished two morphologically distinct types of EGFP positive cells, one expressing glutamate transporters and the other expressing ionotropic glutamate receptors. None of the EGFP-positive cells coexpressed glutamate receptors and transporters. Subpopulations of glutamate receptor bearing EGFP-positive cells expressed AN2, the mouse homolog of the rat NG2 proteoglycan or transcripts for excitatory amino acid carrier 1, a neuronal glutamate transporter. Our data demonstrate the presence of distinct, independent populations of cells with astroglial properties in the developing hippocampus that can differently modulate neuronal signaling pathways. The observed heterogeneity of cells with GFAP promoter-regulated EGFP expression and S100beta/GFAP immunoreactivity challenges the hitherto accepted definition of astrocytes. PMID- 12629180 TI - Oligodendrocyte progenitor enrichment in the connexin32 null-mutant mouse. AB - Before the establishment of chemical synapses, neural progenitors are often coupled by connexin-mediated gap junctions providing a robust mechanism for cell cell communication in developing brain. The present study was undertaken to determine whether alterations in junctional coupling also affect neural progenitor proliferation, survival, and differentiation in adult brain. We localized the connexin32 gap junction protein to a subset of NG2+ and platelet derived growth factor alpha receptor+ early oligodendrocyte progenitors in the dentate gyrus of adult mice. In connexin32-deficient mice, we found an increase in the total number of proliferating nestin+ and NG2+ progenitors in the subgranular zone, hilus, and polymorphonuclear layer of the dentate gyrus in vivo and in the total number of nestin+ progenitors capable of clonogenic expansion in vitro. By bromodeoxyuridine labeling, lineage analysis, and terminal deoxynucleotidyl nick end labeling, we demonstrate that turnover of these cells is constitutively enhanced in the connexin32 knock-out dentate gyrus reflecting a dynamic defect in oligodendrogenesis in this population. Analyses of surviving bromodeoxyuridine-labeled cells at 1, 3, 7, and 28 d after injection demonstrate that this transient amplifying population fails to terminally differentiate and is deleted by an apoptotic-like mechanism within 3 d of labeling. These data provide empirical evidence to support the hypothesis that connexin expression influences adult progenitor number and specifically implicate connexin32-mediated signaling in the activation, survival, and differentiation of a subset of early oligodendrocyte progenitors in postnatal brain. PMID- 12629182 TI - Sensory neuron subtypes have unique substratum preference and receptor expression before target innervation. AB - The factors controlling the specification and subsequent differentiation of sensory neurons are poorly understood. Data from embryological manipulations suggest that either sensory neuron fates are specified by the targets they encounter or sensory neurons are considerably more "plastic" with respect to specification than are neurons of the CNS. The prevailing view that sensory neurons are specified late in development is not consistent, however, with the directed outgrowth of sensory neurons to their targets and the characteristic spatial distribution of sensory neuron fates within the peripheral ganglia. To address when in development different classes of sensory neurons can first be distinguished, we investigated the interactions of early dorsal root ganglia neurons with the extracellular matrix before neurite outgrowth to targets. We found that subclasses of sensory neurons in early dorsal root ganglia show different patterns of neurite outgrowth and integrin expression that are predictive of their fates. In the absence of neurotrophins, presumptive proprioceptive neurons extend neurites robustly on both laminin and fibronectin, whereas presumptive cutaneous neurons show a strong preference for laminin. Cutaneous afferents that have innervated targets show a similar strong preference for laminin and show higher levels of integrin alpha7beta1 than do proprioceptive neurons. Finally, presumptive proprioceptive neurons express fibronectin receptors, integrin alpha3beta1, alpha4beta1, and alpha5beta1, at higher levels than do presumptive cutaneous neurons. Our results indicate that subtypes of sensory neurons have unique patterns of neurite outgrowth and receptor expression before target innervation. PMID- 12629181 TI - Widespread defects in the primary olfactory pathway caused by loss of Mash1 function. AB - MASH1, a basic helix-loop-helix transcription factor, is widely expressed by neuronal progenitors in the CNS and PNS, suggesting that it plays a role in the development of many neural regions. However, in mice lacking a functional Mash1 gene, major alterations have been reported in only a few neuronal populations; among these is a generalized loss of olfactory receptor neurons of the olfactory epithelium. Here, we use a transgenic reporter mouse line, in which the cell bodies and growing axons of subsets of central and peripheral neurons are marked by expression of a tau-lacZ reporter gene (the Tattler-4 allele), to look both more broadly and deeply at defects in the nervous system of Mash1-/- mice. In addition to the expected lack of olfactory receptor neurons in the main olfactory epithelium, developing Mash1-/-;Tattler-4+/- mice exhibited reductions in neuronal cell number in the vomeronasal organ and in the olfactory bulb; the morphology of the rostral migratory stream, which gives rise to olfactory bulb interneurons, was also abnormal. Further examination of cell proliferation, cell death, and cell type-specific markers in Mash1-/- animals uncovered parallels between the main olfactory epithelium and the vomeronasal organ in the regulation of sensory neuron development. Interestingly, this analysis also revealed that, in the olfactory epithelium of Mash1-/- animals, there is an overproduction of proliferating cells that co-express markers of both neuronal progenitors and supporting cells. This finding suggests that olfactory receptor neurons and olfactory epithelium supporting cells may share a common progenitor, and that expression of Mash1 may be an important factor in determining whether these progenitors ultimately generate neurons or glia. PMID- 12629183 TI - Leukemia inhibitory factor is a key signal for injury-induced neurogenesis in the adult mouse olfactory epithelium. AB - The mammalian olfactory epithelium (OE) is composed of primary olfactory sensory neurons (OSNs) that are renewed throughout adulthood by local, restricted neuronal progenitor cells. The molecular signals that control this neurogenesis in vivo are unknown. Using olfactory bulb ablation (OBX) in adult mice to trigger synchronous mitotic stimulation of neuronal progenitors in the OE, we show the in vivo involvement of a cytokine in the cellular events leading to the regeneration of the OE. We find that, of many potential mitogenic signals, only leukemia inhibitory factor (LIF) is induced before the onset of neuronal progenitor proliferation. The rise in LIF mRNA expression peaks at 8 hr after OBX, and in situ RT-PCR and immunocytochemistry indicate that LIF is upregulated, in part, in the injured neurons themselves. This rise in LIF is necessary for injury-induced neurogenesis, as OBX in the LIF knock-out mouse fails to stimulate cell proliferation in the OE. Moreover, delivery of exogenous LIF to the intact adult OE using an adenoviral vector stimulates BrdU labeling in the apical OE. Taken together, these results suggest that injured OSNs release LIF as a stimulus to initiate their own replacement. PMID- 12629184 TI - Redistribution of syntaxin mRNA in neuronal cell bodies regulates protein expression and transport during synapse formation and long-term synaptic plasticity. AB - Syntaxin has an important role in regulating vesicle docking and fusion essential for neurotransmitter release. Here, we demonstrate that the distribution of syntaxin mRNA in cell bodies of sensory neurons (SNs) of Aplysia maintained in cell culture is affected by synapse formation, synapse stabilization, and long term facilitation (LTF) produced by 5-HT. The distribution of the mRNA in turn regulates expression and axonal transport of the protein. Syntaxin mRNA and protein accumulated at the axon hillock of SNs during the initial phase of synapse formation. Significant numbers of granules containing syntaxin were detected in the SN axon. When synaptic strength was stable, both mRNA and protein were targeted away from the axon hillock, and the number of syntaxin granules in the SN axon was reduced. Dramatic increases in mRNA and protein accumulation at the axon hillock and number of syntaxin granules in the SN axon were produced when cultures with stable connections were treated with 5-HT that evoked LTF. Anisomycin (protein synthesis inhibitor) or KT5720 (protein kinase A inhibitor) blocked LTF, accumulation of syntaxin mRNA and protein at the axon hillock, and the increase in syntaxin granules in SN axons. The results indicate that without significant effects on overall mRNA expression, both target interaction and 5-HT via activation of protein kinase A pathway regulate expression of syntaxin and its packaging for transport into axons by influencing the distribution of its mRNA in the SN cell body. PMID- 12629186 TI - Homeostatic effects of depolarization on Ca2+ influx, synaptic signaling, and survival. AB - Depolarization promotes neuronal survival through moderate increases in Ca(2+) influx, but the effects of survival-promoting depolarization (vs conventional trophic support) on neuronal signaling are poorly characterized. We found that chronic, survival-promoting depolarization, but not conventional trophic support, selectively decreased the somatic Ca(2+) current density in hippocampal and cerebellar granule neurons. Depolarization rearing depressed multiple classes of high-voltage activated Ca(2+) current. Consistent with the idea that these changes also affected synaptic Ca(2+) channels, chronic depolarization presynaptically depressed hippocampal neurotransmission. Six days of depolarization rearing completely abolished glutamate transmission but altered GABA transmission in a manner consistent with the alterations of Ca(2+) current. The continued survival of depolarization-reared neurons was extremely sensitive to the re-establishment of basal culture conditions and was correlated with the effects on intracellular Ca(2+) concentration. Thus, compared with cells reared on conventional trophic factors, depolarization evokes homeostatic changes in Ca(2+) influx and signaling that render neurons vulnerable to cell death on activity reduction. PMID- 12629185 TI - Calcitonin gene-related peptide regulates expression of neurokinin1 receptors by rat spinal neurons. AB - Although neurokinin 1 (NK1) receptors contribute to hyperalgesia, and their expression is increased in the spinal cord during peripheral inflammation, little is known regarding the signaling molecules and the second messenger pathways that they activate in regulating the expression of the NK1 receptor gene. Because the promoter region of the NK1 receptor contains a cAMP response element (CRE), we tested the hypothesis that calcitonin gene-related peptide (CGRP) regulates the expression of NK1 receptors via a pathway involving activation of the transcription factor cAMP response element binding protein (CREB). Experiments were conducted on primary cultures of neonatal rat spinal neurons. Treatment of cultures with CGRP for 8-24 hr increased (125)I-substance P binding on spinal neurons; the increase in binding was preceded by an elevation in NK1 receptor mRNA. The CGRP-induced change in (125)I-substance P binding was concentration dependent and was inhibited by the antagonist CGRP(8-37). CGRP increased phosphorylated CREB immunoreactivity and CRE-dependent transcription in neurons, indicating the involvement of the transcription factor CREB. Evidence that CGRP increased cAMP levels in spinal neurons and that the protein kinase A inhibitor H89 attenuated CGRP-induced CRE-dependent transcription suggests that the intracellular pathway stimulated by CGRP leads to activation of protein kinase A. Collectively these data define a role for CGRP as a signaling molecule that induces expression of NK1 receptors in spinal neurons. The data provide evidence that a neuropeptide receptor controls gene expression in the CNS and add another dimension to understanding the cotransmission of substance P and CGRP by primary afferent neurons. PMID- 12629187 TI - Perinatal neurosteroid levels influence GABAergic interneuron localization in adult rat prefrontal cortex. AB - Neurosteroids are a class of steroids synthesized de novo in the brain, several of which are potent modulators of GABA(A) receptor function. In developing brain GABA(A) receptor, stimulation plays a trophic role. Cortical levels of the GABAergic neurosteroid 3alpha-hydroxy-5alpha-pregnan-20-one (3alpha,5alpha-THP) vary dramatically across development; during the second week of life, elevated levels of 3alpha,5alpha-THP are associated with decreased GABA(A) receptor function. To determine whether alteration of endogenous 3alpha,5alpha-THP levels during development alters GABAergic interneurons in prefrontal cortex (PFC) at maturity, rat pups were exposed to 3alpha,5alpha-THP (10 mg/kg) on postnatal day 1 (P1), P2, and P5. On P80, frontal cortex tissue was assayed for GABAergic cell localization (parvalbumin and calbindin immunoreactivity), agonist-dependent [(3)H] dizocilpine (MK-801) binding to NMDA receptors in cortical homogenates, muscimol-mediated (36)Cl(-) influx into synaptoneurosomes, and 3alpha,5alpha-THP levels. The localization of parvalbumin-labeled cells was markedly altered; the ratio of cell number in the deep layers (V-VI) versus superficial layers (I-III) of adult PFC increased twofold in animals exposed to 3alpha,5alpha-THP on P1 or P5. Relative microtubule-associated protein-2 and calbindin immunoreactivity were not altered by perinatal 3alpha,5alpha-THP administration. Agonist-dependent [(3)H]MK-801 binding was decreased in PFC but not parietal cortex homogenates, whereas muscimol-mediated (36)Cl(-) influx and 3alpha,5alpha-THP levels were unchanged in frontal cortex of adult males exposed to 3alpha,5alpha-THP on P5. These data are consistent with a change in the distribution of a subset of interneurons in response to neurosteroid exposure and suggest that GABAergic neurosteroids are critical for normal development of GABAergic systems in the PFC. PMID- 12629188 TI - Excitatory actions of endogenously released GABA contribute to initiation of ictal epileptiform activity in the developing hippocampus. AB - In the developing rat hippocampus, ictal epileptiform activity can be elicited easily in vitro during the first three postnatal weeks. Changes in neuronal ion transport during this time cause the effects of GABA(A) receptor (GABA(A)-R) activation to shift gradually from strongly depolarizing to hyperpolarizing. It is not known whether the depolarizing effects of GABA and the propensity for ictal activity are causally linked. A key question is whether the GABA-mediated depolarization is excitatory, which we defined operationally as being sufficient to trigger action potentials. We assessed the effect of endogenous GABA on ictal activity and neuronal firing rate in hippocampal slices from postnatal day 1 (P1) to P30. In extracellular recordings, there was a strong correlation between the postnatal age at which GABA(A)-R antagonists decreased action potential frequency (P23) and the age at which ictal activity could be induced by elevated potassium (P23). In addition, there was a strong correlation between the fraction of slices in which ictal activity was induced by elevated potassium concentrations and the fractional decrease in action potential firing when GABA(A)-Rs were blocked in the presence of ionotropic glutamate receptor antagonists. Finally, ictal activity induced by elevated potassium was blocked by the GABA(A)-R antagonists bicuculline and SR-95531 (gabazine) and increased in frequency and duration by GABA(A)-R agonists isoguvacine and muscimol. Thus, the propensity of the developing hippocampus for ictal activity is highly correlated with the effect of GABA on action potential probability and reversed by GABA(A) antagonists, indicating that GABA-mediated excitation is causally linked to ictal activity in this developmental window. PMID- 12629190 TI - Optical imaging of long-term depression in the mouse cerebellar cortex in vivo. AB - Conjunctive stimulation of climbing fiber and parallel fiber inputs results in long-term depression (LTD) at parallel fiber-Purkinje cell synapses. Although hypothesized to play a major role in cerebellar motor learning, there has been no characterization of the cellular and molecular mechanisms of LTD in the whole animal, let alone its spatial properties, both of which are critical to understanding the role of LTD in cerebellar function. Neutral red optical imaging of the cerebellar cortex in the anesthetized mouse was used to visualize the spatial patterns of activation. Stimulation of the parallel fibers evoked a transverse beam of optical activity, and stimulation of the contralateral inferior olive evoked parasagittal bands. Conjunctive stimulation of parallel fibers and climbing fibers induced a long-term decrease (at least 1 hr) in the optical response to subsequent parallel fiber activation confined to the region of interaction between these two inputs. Activation of climbing fibers alone failed to induce the long-term decrease. Field potential recordings confirmed that the depression is postsynaptic and restricted to the interaction site. The long-term depression in the beam was prevented by a group 1 metabotropic glutamate receptor (mGluR(1)) antagonist and was absent in transgenic mice selectively expressing an inhibitor of protein kinase C (PKC) in Purkinje cells. Conversely, the long-term depression occurred in the mGluR(4) knock-out mouse, consistent with its postsynaptic origin. In addition to providing the first visualization of parallel fiber-Purkinje cell LTD in the cerebellar cortex, this study demonstrates the spatial specificity of LTD and its dependence on mGluR(1) and PKC in vivo. PMID- 12629189 TI - Peptidases prevent mu-opioid receptor internalization in dorsal horn neurons by endogenously released opioids. AB - To evaluate the effect of peptidases on mu-opioid receptor (MOR) activation by endogenous opioids, we measured MOR-1 internalization in rat spinal cord slices. A mixture of inhibitors of aminopeptidases (amastatin), dipeptidyl carboxypeptidase (captopril), and neutral endopeptidase (phosphoramidon) dramatically increased the potencies of Leu-enkephalin and dynorphin A to produce MOR-1 internalization, and also enhanced the effects of Met-enkephalin and alpha neoendorphin, but not endomorphins or beta-endorphin. The omission of any one inhibitor abolished Leu-enkephalin-induced internalization, indicating that all three peptidases degraded enkephalins. Amastatin preserved dynorphin A-induced internalization, and phosphoramidon, but not captopril, increased this effect, indicating that the effect of dynorphin A was prevented by aminopeptidases and neutral endopeptidase. Veratridine (30 microm) or 50 mm KCl produced MOR-1 internalization in the presence of peptidase inhibitors, but little or no internalization in their absence. These effects were attributed to opioid release, because they were abolished by the selective MOR antagonist CTAP (D-Phe Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH(2)) and were Ca(2+) dependent. The effect of veratridine was protected by phosphoramidon plus amastatin or captopril, but not by amastatin plus captopril or by phosphoramidon alone, indicating that released opioids are primarily cleaved by neutral endopeptidase, with a lesser involvement of aminopeptidases and dipeptidyl carboxypeptidase. Therefore, because the potencies of endomorphin-1 and endomorphin-2 to elicit internalization were unaffected by peptidase inhibitors, the opioids released by veratridine were not endomorphins. Confocal microscopy revealed that MOR-1-expressing neurons were in close proximity to terminals containing opioids with enkephalin-like sequences. These findings indicate that peptidases prevent the activation of extrasynaptic MOR-1 in dorsal horn neurons. PMID- 12629191 TI - Neuronal basis of crossed actions from the reticular formation on feline hindlimb motoneurons. AB - Pathways through which reticulospinal neurons can influence contralateral limb movements were investigated by recording from motoneurons innervating hindlimb muscles. Reticulospinal tract fibers were stimulated within the brainstem or in the lateral funiculus of the thoracic spinal cord contralateral to the motoneurons. Effects evoked by ipsilaterally descending reticulospinal tract fibers were eliminated by a spinal hemisection at an upper lumbar level. Stimuli applied in the brainstem evoked EPSPs, IPSPs, or both at latencies of 1.42 +/- 0.03 and 1.53 +/- 0.04 msec, respectively, from the first components of the descending volleys and with properties indicating a disynaptic linkage, in most contralateral motoneurons: EPSPs in 76% and IPSPs in 26%. EPSPs with characteristics of monosynaptically evoked responses, attributable to direct actions of crossed axon collaterals of reticulospinal fibers, were found in a small proportion of the motoneurons, whether evoked from the brainstem (9%) or from the thoracic cord (12.5%). Commissural neurons, which might mediate the crossed disynaptic actions (i.e., were antidromically activated from contralateral motor nuclei and monosynaptically excited from the ipsilateral reticular formation), were found in Rexed's lamina VIII in the midlumbar segments (L3-L5). The results reveal that although direct actions of reticulospinal fibers are much more potent on ipsilateral motoneurons, interneuronally mediated actions are as potent contralaterally as ipsilaterally, and midlumbar commissural neurons are likely to contribute to them. They indicate a close coupling between the spinal interneuronal systems used by the reticulospinal neurons to coordinate muscle contractions ipsilaterally and contralaterally. PMID- 12629192 TI - Norepinephrine in the prefrontal cortex is critical for amphetamine-induced reward and mesoaccumbens dopamine release. AB - Increasing evidence points to a major involvement of cortical areas in addictive mechanisms. Noradrenergic transmission in the medial prefrontal cortex (mpFC) has been shown to affect the motor effects of amphetamine, although there is no evidence of its involvement in the rewarding effects of this psychostimulant. The present experiments were aimed at investigating the possibility of a selective involvement of prefrontal cortical norepinephrine (NE) in the rewarding reinforcing effects of amphetamine. To do so, we evaluated the effects of mpFC NE selective depletion in mice of C57BL/6J inbred strain, a background commonly used in molecular approaches that is known to be highly susceptible to the rewarding effects of the psychostimulant. In a first set of experiments, we demonstrated the absence of amphetamine-induced conditioned place preference in mice bearing prefrontal NE depletion. In a second series of experiments, we demonstrated that the same lesion dramatically reduced amphetamine-induced mesoaccumbens dopamine release as measured by intracerebral microdialysis. These results indicate that noradrenergic prefrontal transmission, by allowing increased dopamine release in the nucleus accumbens induced by amphetamine, is a critical factor for the rewarding-reinforcing effects of this drug. PMID- 12629193 TI - Task-dependent presynaptic inhibition. AB - This study compares the level of presynaptic inhibition during two rhythmic movements in the cat: locomotion and scratch. Dorsal rootlets from L6, L7, or S1 segments were cut, and their proximal stumps were recorded during fictive locomotion occurring spontaneously in decerebrate cats and during fictive scratch induced by d-tubocurarine applied on the C1 and C2 segments. Compared with rest, the number of antidromic spikes was increased (by 12%) during locomotion, whereas it was greatly decreased (31%) during scratch, and the amplitude of dorsal root potentials (DRPs), evoked by stimulating a muscle nerve, was slightly decreased (7%) during locomotion but much more so during scratch (53%). When compared with locomotion, the decrease in the number of antidromic spikes (45%) and the decrease in DRP amplitude (43%) during scratch were of similar magnitude. Also, the amplitude of primary afferent depolarization (PAD), recorded with micropipettes in axons (n = 13) of two cats, was found to be significantly reduced (60%) during scratch compared with rest. During both rhythms, there were cyclic oscillations in dorsal root potential the timing of which was linearly related to the timing of rhythmic activity in tibialis anterior. The amplitude of these oscillations was significantly smaller (34%) during locomotion compared with scratch. These results suggest that the reduction in antidromic activity during scratch was attributable to a task-dependent decrease in transmission in PAD pathways and not to underlying potential oscillations related to the central pattern generator. It is concluded that presynaptic inhibition and antidromic discharge may have a more important role in the control of locomotion than scratch. PMID- 12629194 TI - Localized lesion of caudal part of lobus parolfactorius caused impulsive choice in the domestic chick: evolutionarily conserved function of ventral striatum. AB - Effects of bilateral chemical lesions of the medial basal ganglia [lobus parolfactorius (LPO)] were examined in 7- to 14-d-old domestic chicks. Chicks were trained in a color discrimination task, in which the subject had to peck one of the two colored beads associated with rewards that differed in quantity (amount of food) and/or temporal proximity (delay of food delivery from peck). In experiment 1, food was given without delay, and chicks successfully learned to choose a colored bead that was associated with a larger reward than the other. In experiment 2, a colored bead (red) was associated with a large reward delivered after a delay (D = 1, 2, or 3 sec), whereas another (yellow) was associated with a small reward delivered immediately. In intact and sham-operated conditions, chicks with a longer D chose the red bead progressively fewer times. Selective lesions to the caudal LPO (but not the rostral LPO) caused impulsive choice, and the ablated chicks chose the yellow bead and gained a small-immediate reward regardless of D. However, when retrained in a null-delay condition (D = 0 sec), the lesioned chick chose the red bead again. Ability to associate novel colors with reward was also unimpaired. These results suggest that the LPO may be responsible for the anticipation of reward proximity and involved in a suppression of impulsiveness by which animals seek immediate gains. The present results also indicate a striking similarity in functional roles between the avian LPO and the nucleus accumbens/ventral striatum in mammals. PMID- 12629195 TI - Representation of odor habituation and timing in the hippocampus. AB - We performed simultaneous single-neuron recordings from the hippocampus and the olfactory bulb of anesthetized, freely breathing rats. Odor response properties of neurons in the olfactory bulb and hippocampus were characterized as firing rate changes or respiration-coupled changes. A panel of five odors was used. The rats had not been exposed to the odors on the panel before the experiment. The olfactory bulb and hippocampal neurons responded to repeated odor presentations in two ways: first, by changes in firing rate, and second, by respiratory tuning changes. Approximately 60% of bulbar neurons, 48% of hippocampal CA1 neurons, and 12% of hippocampal CA3 neurons showed statistically significant responses. None of the odor-responsive neurons in either the bulb or hippocampus responded to all of the odors on the panel. Repeated 10 sec odor stimuli presented at the intervals of 20, 30, 60, 110, and 160 sec were used to analyze the effect of the interval on odor response properties of the recorded neurons. Bulbar neurons were relatively nonselective for odor interval. Hippocampal neurons showed unexpected selectivity for the interval between repeated odor presentations. CA1 and CA3 neurons responded to only one to three of the intervals in the range. On the basis of these findings, we postulate that the hippocampus has the ability to keep track of the time elapsed between consecutive odor stimuli. This may act as a neuronal substrate for habituation and for complex tasks such as odor-guided navigation. PMID- 12629196 TI - Stimulation of the subthalamic nucleus changes the firing pattern of pallidal neurons. AB - To clarify the mechanism underlying improvement of parkinsonian signs by high frequency electrical stimulation (HFS) of the subthalamic nucleus (STN), we investigated the effects of STN HFS on neuronal activity of the internal and external segment of the globus pallidus (GPi and GPe, respectively) in two rhesus monkeys rendered parkinsonian by administration of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine. A scaled-down version of the chronic stimulating electrode used in humans, consisting of four metal contacts 0.50 mm in length each separated by 0.50 mm, was implanted through a cephalic chamber targeting the STN. Histological reconstruction revealed that the cathode was located in the STN in both monkeys. Extracellular recordings from a total of 110 pallidal neurons during STN stimulation were performed. Poststimulus time histograms of single neurons triggered by 2 Hz STN stimulation pulses at 2.4-3.0 V revealed short latency excitations at 2.5-4.5 and 5.5-7.0 msec after stimulation onset and inhibitions at 1.0-2.5, 4.5-5.5, and 7.0-9.0 msec for both GPe and GPi neurons. These short-latency responses were present with 136 Hz stimulation, at voltages effective for alleviation of parkinsonian signs, resulting in a significant increase in mean discharge rate and a stimulus-synchronized regular firing pattern. These results indicate that activation of the STN efferent fibers and resultant changes in the temporal firing pattern of neurons in GPe and GPi underlie the beneficial effect of HFS in the STN in Parkinson's disease and further support the role of temporal firing patterns in the basal ganglia in the development of Parkinson's disease and other movement disorders. PMID- 12629197 TI - Reversible disorganization of the locomotor pattern after neonatal spinal cord transection in the rat. AB - The central pattern generators (CPGs) for locomotion, located in the lumbar spinal cord, are functional at birth in the rat. Their maturation occurs during the last few days preceding birth, a period during which the first projections from the brainstem start to reach the lumbar enlargement of the spinal cord. The goal of the present study was to investigate the effect of suppressing inputs from supraspinal structures on the CPGs, shortly after their formation. The spinal cord was transected at the thoracic level at birth [postnatal day 0 (P0)]. We examined during the first postnatal week the capacity of the CPGs to produce rhythmic motor activity in two complementary experimental conditions. Left and right ankle extensor muscles were recorded in vivo during airstepping, and lumbar ventral roots were recorded in vitro during pharmacologically evoked fictive locomotion. Mechanical stimulation of the tail elicited long-lasting sequences of airstepping in the spinal neonates and only a few steps in sham-operated rats. In vitro experiments made simultaneously on spinal and sham animals confirmed the increased excitability of the CPGs after spinalization. A left-right alternating locomotor pattern was observed at P1-P3. Both types of experiments showed that the pattern was disorganized at P6-P7, and that the left-right alternation was lost. Alternation was restored after the activation of serotonergic 5-HT(2) receptors in vivo. These results suggest that descending pathways, in particular serotonergic projections, control the strength of reciprocal inhibition and therefore shape the locomotor pattern in the neonatal rat. PMID- 12629198 TI - Discharge of raphe magnus ON and OFF cells is predictive of the motor facilitation evoked by repeated laser stimulation. AB - Medullary raphe magnus (RM) on and off cells are thought to modulate spinal nociception by gating withdrawals evoked by noxious stimulation. To test whether withdrawal initiation is the target of RM modulation, we examined the relationship between on and off cell discharge and motor withdrawal evoked by noxious laser heat in halothane-anesthetized rats. The cellular responses of both cell types began during the 50 msec after onset of the tail flick, peaked within 200 msec, and outlasted the duration of the motor reaction. Thus, it is unlikely that the target of on and off cell modulation is withdrawal initiation; instead, on and off cells may modulate reactions to repeated noxious stimulation. We therefore tested whether laser heat-evoked changes in RM cell discharge were predictive of the modulatory effects of one noxious stimulus on the reaction to a subsequent noxious stimulus. Two pulses of laser heat were presented at interpulse intervals of 0.8, 2.0, or 10.0 sec. The motor withdrawal evoked by the second pulse was significantly enhanced relative to that evoked by the first pulse. The observed motor enhancement depended on supraspinal input because it was not present in spinalized rats. Comparison of the relative changes in motor and cellular activity preceding double laser heat stimulation revealed parallel changes between motor facilitation, decreases in off cell discharge, and increases in on cell discharge. This finding suggests a preparatory role for RM on and off cells in enhancing reactions to a noxious stimulus that closely follows another noxious stimulus. PMID- 12629199 TI - On the role of nitric oxide in hippocampal long-term potentiation. AB - Nitric oxide (NO) functions in several types of synaptic plasticity, including hippocampal long-term potentiation (LTP), in which it may serve as a retrograde messenger after postsynaptic NMDA receptor activation. In accordance with a prediction of this hypothesis, and with previous findings using guinea pig tissue, exogenous NO, when paired with a short tetanus (ST) to afferent fibers, generated a stable NMDA receptor-independent potentiation of rat CA1 hippocampal synaptic transmission that occluded LTP. Contrary to predictions, however, the pairing-induced potentiation was abolished in the presence of NO synthase inhibitors, indicating that endogenous NO is required for exogenous NO to facilitate LTP. Periodic application of NO while endogenous NO synthesis was blocked indicated that a tonic low level is necessary on both sides of the NO-ST pairing for the plasticity to occur. A similar dependence on tonic NO seems to extend to LTP, because application of an NO synthase inhibitor 5 min after tetanic stimulation blocked LTP as effectively as adding it beforehand. The posttetanus time window during which NO operated was restricted to <15 min. Inhibition of the guanylyl cyclase-coupled NO receptor indicated that the potentiation resulting from NO-ST pairing and the NO signal transduction pathway during early LTP are both through cGMP. We conclude that NO does not function simply as an acute signaling molecule in LTP induction but has an equally important role outside this phase. The results resonate with observations concerning the role of the hippocampal NO-cGMP pathway in certain types of learning behavior. PMID- 12629200 TI - Serum transthyretin monomer as a possible marker of blood-to-CSF barrier disruption. AB - The CNS is shielded from systemic influences by two separate barriers, the blood brain barrier (BBB) and the blood-to-CSF barrier. Failure of either barrier bears profound significance in the etiology and diagnosis of several neurological diseases. Furthermore, selective opening of BBB tight junctions provides an opportunity for delivery of otherwise BBB impermeant drugs. Peripheral assessment of BBB opening can be achieved by detection in blood of brain-specific proteins that extravasate when these endothelial junctions are breached. We developed a proteomic approach to discover clusters of CNS-specific proteins with extravasation into serum that correlates with BBB openings. Protein profiles from blood samples obtained from patients undergoing iatrogenic BBB disruption (BBBD) with intra-arterial hyperosmotic mannitol were compared with pre-BBB opening serum. A low molecular weight protein (14 kDa) identified by mass spectroscopy as transthyretin (TTR) consistently correlated with BBBD. Protein gel electrophoresis and immunodetection confirmed that TTR was indeed extravasated in its monomeric form when CNS barriers were breached. The time course of TTR extravasation was compared with release from the brain of another BBB integrity marker, S-100beta (11 kDa). Kinetic analysis revealed that the appearance of S 100beta, presumably originating from perivascular astrocytic end feet, preceded extravasation of TTR by several minutes. Because TTR is localized primarily in choroid plexus and, as a soluble monomer, in CSF, we concluded that although S 100beta is a marker of BBB integrity, TTR instead may be a peripheral tracer of blood-to-cerebrospinal barrier. PMID- 12629202 TI - Feedback inhibition defines transverse processing modules in the lateral amygdala. AB - The lateral amygdaloid (LA) nucleus is the main input station of the amygdala for sensory afferents. However, it is unclear how the lateral nucleus transforms these inputs, because its intrinsic connectivity is poorly understood. Here, we took advantage of the fact that glutamatergic neurons of the lateral nucleus send a primarily unidirectional projection to the basomedial nucleus. Consequently, it was possible to infer the targets of their intranuclear axons (projection cells vs inhibitory interneurons) by backfiring some projection neurons from the basomedial nucleus and analyzing evoked responses in other LA projection cells. Basomedial stimuli evoked markedly different synaptic responses depending on the orientation of the slices. In coronal slices (intact and decorticated), the prevalent response of LA neurons was an inhibition, regardless of the stimulation intensity. This inhibition was sensitive to GABA(A) and non-NMDA receptor antagonists, suggesting that it was mediated by the activation of GABAergic cells of the LA. In contrast, basomedial stimuli primarily evoked EPSPs in horizontal slices, regardless of the position of recorded neurons. In light of these findings, we conclude that the prevalent target of the intrinsic axon collaterals of projection cells depend on the rostrocaudal position of target neurons with respect to the parent cell body: inhibitory interneurons at rostrocaudal proximity versus other projection cells at a distance. Thus, feedback interneurons effectively divide the lateral nucleus in transverse processing modules that prevent runaway excitation within each module but allow intermixing of sensory information in the rostrocaudal plane. PMID- 12629201 TI - One-trial memory for object-place associations after separate lesions of hippocampus and posterior parahippocampal region in the monkey. AB - In earlier studies of one-trial spatial memory in monkeys (Parkinson et al., 1988; Angeli et al., 1993), severe and chronic memory impairment for both object place association and place alone was found after ablation of the hippocampal formation. The results appeared to provide the first clear-cut evidence in the monkey of the essential role of the hippocampus in spatial memory, but that interpretation neglected the inclusion in the lesion of the underlying posterior parahippocampal region. To determine the separate contributions of the hippocampus and posterior parahippocampal region to these spatial forms of one trial memory, we trained 10 rhesus monkeys, as before, to remember the spatial positions of either two different trial-unique objects overlying two of the wells in a three-well test tray (object-place trials) or simply two of the three wells (place trials). Six of the monkeys then received ibotenic acid lesions restricted to the hippocampal formation (group H), and the four others received selective ablations of the posterior parahippocampal region (group P), comprising mainly parahippocampal cortex, parasubiculum, and presubiculum. Group H was found to be completely unaffected postoperatively on both types of trials, whereas group P sustained an impairment on both types equal in magnitude to that observed after the combined lesions in the original studies. Thus, contrary to the previous interpretation, one-trial memory for object-place association and, perhaps more fundamentally, one-trial memory for two different places appear to be critically dependent not on the hippocampal formation but rather on the posterior parahippocampal region. PMID- 12629204 TI - Sensorimotor memory for fingertip forces: evidence for a task-independent motor memory. AB - When repetitively lifting an object with randomly varying mechanical properties, the fingertip forces reflect the previous lift. We examined the specificity of this "sensorimotor memory" by observing the effects of an isolated pinch on the subsequent lift of a known object. In this case, the pinch force was unrelated to the fingertip forces necessary to grip the object efficiently. The peak grip force used to lift the test object (4 N weight) depended on the preceding task. Compared with repetitively lifting the 4 N test object, the peak grip force was 2 N greater when a lift of the same object was preceded by a lift in which a hidden mass was attached to the object to increase the weight to 8 N. This 2 N increase in grip force also occurred when subjects lifted the 4 N test object after pinching a force transducer with a force of 8 N. Thus, similar grip forces were stored in sensorimotor memory for both tasks, and reflected subjects' use of 7.9 +/- 1.1 N to lift the 8 N object. Similar effects occurred when the preceding pinch or lift was performed with the opposite hand. The peak lift force was unaffected by the isolated pinch, suggesting that a generalized increase in fingertip and limb forces did not occur. We conclude that the sensorimotor memory is not specific for lifting an object. It is doubtful that this particular memory stores the physical properties of objects or reflects a forward internal model for predictively controlling fingertip forces. PMID- 12629203 TI - Glial cell line-derived neurotrophic factor increases stimulus-evoked dopamine release and motor speed in aged rhesus monkeys. AB - Changes in the functional dynamics of dopamine release and regulation in the basal ganglia have been posited to contribute to age-related slowing of motor functions. Here, we report the effects of glial cell line-derived neurotrophic factor (GDNF) on the stimulus-evoked release of dopamine and motor speed in aged monkeys (21-27 years of age; n = 10). Although no changes were observed in the vehicle controls (n = 5), chronic infusions of 7.5 microg of GDNF per day for 2 months into the right lateral ventricle initially increased hand movement speed up to 40% on an automated hand-reach task. These effects were maintained for at least 2 months after replacing GDNF with vehicle, and increased up to another 10% after the reinstatement of GDNF treatment for 1 month. In addition, upper-limb motor performance times of the aged GDNF-treated animals (n = 5) recorded at the end of the study were similar to those of five young adult monkeys (8-12 years of age). The stimulus-evoked release of dopamine was significantly increased, up to 130% in the right caudate nucleus and putamen and up to 116% in both the right and left substantia nigra of the aged GDNF recipients compared with vehicle controls. Also, basal extracellular levels of dopamine were bilaterally increased, up to 163% in the substantia nigra of the aged GDNF-treated animals. The data suggest that the effects of GDNF on the release of dopamine in the basal ganglia may be responsible for the improvements in motor functions and support the hypothesis that functional changes in dopamine release may contribute to motor dysfunctions characterizing senescence. PMID- 12629205 TI - Early growth response 1 protein, an upstream gatekeeper of the p53 tumor suppressor, controls replicative senescence. AB - The proliferation of most primary cells in culture is limited by replicative senescence and crisis, p53-dependent events. However, the regulation of p53 itself has not been defined. We find that deletion of the early growth response 1 (EGR1) transcription factor leads to a striking phenotype, including complete bypass of senescence and apparent immortal growth consistent with loss of a suppressor gene. EGR1-null mouse embryo fibroblasts (MEFs) exhibit decreased expression of p53, p21(Cip1/Waf1), and other p53 "marker" proteins. Precrisis WT but not EGR1-null cells exhibit irradiation-induced arrest. WT MEFs that emerge from crisis exhibit a mutated p53 (sequence confirmed), colony formation, and tumorigenicity. In contrast, high-passage EGR1-null MEFs retain the WT p53 sequence but with much reduced expression, remain untransformed, and grow continuously. An EGR1-expressing retrovirus restores p53 expression and sencescence to EGR1-null but not p53-null MEFs or postcrisis WT cells. Taken together, the results establish EGR1 as a major regulator of cell senescence and previously undescribed upstream "gatekeeper" of the p53 tumor suppressor pathway. PMID- 12629206 TI - Mechanics of DNA packaging in viruses. AB - A new generation of single-molecule experiments has opened up the possibility of reexamining many of the fundamental processes of biochemistry and molecular biology from a unique and quantitative perspective. One technique producing a host of intriguing results is the use of optical tweezers to measure the mechanical forces exerted by molecular motors during key processes such as the transcription of DNA or the packing of a viral genome into its capsid. The objective of the current article is to respond to such measurements on viruses and to use the theory of elasticity and a simple model of charge and hydration forces to derive the force required to pack DNA into a viral capsid as a function of the fraction of the viral genome that has been packed. The results are found to be in excellent accord with recent measurements and complement previous theoretical work. Because the packing of DNA in viral capsids occurs under circumstances of high internal pressure, we also compute how much pressure a capsid can sustain without rupture. PMID- 12629208 TI - Microsecond electrophoresis. AB - Although analysis strategies exist for probing a diverse array of molecular properties, most of these approaches are not amenable to the study of reaction intermediates and other transient species. Separations in particular can provide detailed information on attributes not readily measured by spectroscopy but typically are performed over time scales much longer than the life span of highly unstable compounds. Here we report the development of an electrophoretic strategy that dramatically extends the practical speed limit for fractionations and demonstrate its utility in examining transient hydroxyindole photoproducts. Fluorescent reaction intermediates are optically generated in femtoliter volumes within a flowing reagent stream and are differentially transported at velocities as large as 1.3 m.s(-1), thereby minimizing band variance and allowing multicomponent reaction mixtures to be resolved over separation paths as short as 9 microm. Analyte migration times and band variances do not deviate significantly from basic theory for separations performed with fields that exceed 0.1 MV.cm( 1), indicating that effects from Joule heating are minor. We demonstrate the feasibility of achieving baseline resolution of a binary mixture in <10 micros, nearly 100-fold faster than previously possible. Application of this approach to the study of a range of short-lived molecules should be feasible. PMID- 12629207 TI - The topoisomerase IIbeta circular clamp arrests transcription and signals a 26S proteasome pathway. AB - It has been proposed that the topoisomerase II (TOP2)beta-DNA covalent complex arrests transcription and triggers 26S proteasome-mediated degradation of TOP2beta. It is unclear whether the initial trigger for proteasomal degradation is due to DNA damage or transcriptional arrest. In the current study we show that the TOP2 catalytic inhibitor 4,4-(2,3-butanediyl)-bis(2,6-piperazinedione) (ICRF 193), which traps TOP2 into a circular clamp rather than the TOP2-DNA covalent complex, can also arrest transcription. Arrest of transcription, which is TOP2beta-dependent, is accompanied by proteasomal degradation of TOP2beta. Different from TOP2 poisons and other DNA-damaging agents, ICRF-193 did not induce proteasomal degradation of the large subunit of RNA polymerase II. These results suggest that proteasomal degradation of TOP2beta induced by the TOP2-DNA covalent complex or the TOP2 circular clamp is due to transcriptional arrest but not DNA damage. By contrast, degradation of the large subunit of RNA polymerase II is due to a DNA-damage signal. PMID- 12629209 TI - Cost minimization by helpers in cooperative vertebrates. AB - When parents invest heavily in reproduction they commonly suffer significant energetic costs. Parents reduce the long-term fitness implications of these costs through increased foraging and reduced reproductive investment in the future. Similar behavioral modifications might be expected among helpers in societies of cooperative vertebrates, in which helping is associated with energetic costs. By using multivariate analyses and experiments, we show that in cooperative meerkats, Suricata suricatta, helping is associated with substantial short-term growth costs but limited long-term fitness costs. This association forms because individual contributions to cooperation are initially condition dependent, and, because when helpers invest heavily in cooperation, they increase their foraging rate during the subsequent nonbreeding period and reduce their level of cooperative investment in the subsequent reproductive period. These results provide a unique demonstration that despite significant short-term costs, helpers, like breeders, are able to reduce the fitness consequences of these costs through behavioral modifications. PMID- 12629210 TI - Intein-mediated assembly of a functional beta-glucuronidase in transgenic plants. AB - The DnaE intein in Synechocystis sp. strain PCC6803 is the first and only naturally split intein that has been identified so far. It is capable of catalyzing a protein trans-splicing mechanism to assemble a mature protein from two separate precursors. Therefore, it is a powerful tool for protein modification and engineering. Inteins have not been identified, nor have intein mediated protein splicing reactions been demonstrated, in plant cells. In this paper, we describe the use of the Ssp DnaE split intein in transgenic plants for reconstitution of a protein trans-splicing reaction. We have synthesized artificial genes that encode for N-terminal half (Int-n) and C-terminal half (Int c) fragments of Ssp DnaE split intein and divided beta-glucuronidase (GUS) gene to encode GUS-n and GUS-c parts of the enzyme as reporter. The in-frame fusions of GUSn/Intn and Intc/GUSc were constructed and transfected into Arabidopsis. We have observed in vivo reassembly of functional beta-glucuronidase when both GUSn/Intn and Intc/GUSc constructs were introduced into the same Arabidopsis genome either by cotransformation or through genetic crossing, hereby signifying an intein-mediated protein trans-splicing mechanism reconstituted in plant cells. PMID- 12629211 TI - Enhanced insulin sensitivity in mice lacking ganglioside GM3. AB - Gangliosides are sialic acid-containing glycosphingolipids that are present on all mammalian plasma membranes where they participate in recognition and signaling activities. We have established mutant mice that lack GM3 synthase (CMP NeuAc:lactosylceramide alpha2,3-sialyltransferase; EC 2.4.99.-). These mutant mice were unable to synthesize GM3 ganglioside, a simple and widely distributed glycosphingolipid. The mutant mice were viable and appeared without major abnormalities but showed a heightened sensitivity to insulin. A basis for the increased insulin sensitivity in the mutant mice was found to be enhanced insulin receptor phosphorylation in skeletal muscle. Importantly, the mutant mice were protected from high-fat diet-induced insulin resistance. Our results show that GM3 ganglioside is a negative regulator of insulin signaling, making it a potential therapeutic target in type 2 diabetes. PMID- 12629212 TI - Gelsolin is a dorsalizing factor in zebrafish. AB - The gene for gelsolin (an actin-binding, cytoskeletal regulatory protein) was shown earlier to be specialized for high corneal expression in adult zebrafish. We show here that zebrafish gelsolin is required for proper dorsalization during embryogenesis. Inhibition of gelsolin expression by injecting fertilized eggs with a specific morpholino oligonucleotide resulted in a range of concentration dependent ventralized phenotypes, including those lacking a brain and eyes. These were rescued by coinjection of zebrafish gelsolin or chordin (a known dorsalizing agent) mRNAs, or human gelsolin protein. Moreover, injection of gelsolin mRNA or human gelsolin protein by itself dorsalized the developing embryos, often resulting in axis duplication. Injection of the gelsolin-specific morpholino oligonucleotide enhanced the expression of Vent mRNA, a ventral marker downstream of bone morphogenetic proteins, whereas injection of gelsolin mRNA enhanced the expression of chordin and goosecoid mRNAs, both dorsal markers. Our results indicate that gelsolin also modulates embryonic dorsalventral pattern formation in zebrafish. PMID- 12629213 TI - Sequences in the (A)gamma-delta intergenic region are not required for stage specific regulation of the human beta-globin gene locus. AB - The human beta-globin locus has been extensively studied as a model of tissue and developmental stage-specific gene expression. Structural mapping of naturally occurring mutations, including transfection and transgenic studies, and the recent finding of intergenic transcripts have suggested that there are cis-acting sequence elements in the (A)gamma-delta intergenic region involved in regulating gamma- and beta-globin gene expression. To determine whether previously identified sequences in the (A)gamma-delta intergenic region are required for appropriate developmental expression of the human beta-globin gene cluster, transgenic mice were generated by transfer of yeast artificial chromosomes containing the entire human beta-globin locus. Three different deletions of the (A)gamma-delta intergenic region were introduced, including (i) deletion of the 750-bp (A)gamma 3' regulatory element ((A)gammae), (ii) deletion of 3.2 kb upstream of the delta-globin gene encompassing pyrimidine-rich sequences and the recently described intergenic transcript initiation site, and (iii) deletion of a 12.5-kb fragment encompassing most of the (A)gamma-delta globin intergenic region. Analysis of multiple transgenic lines carrying these deletion constructs demonstrated that the normal stage-specific sequential expression of the epsilon , gamma-, and beta-globin genes was preserved, despite deletion of these putative regulatory sequences. These studies suggest that regulatory sequences required for activation and silencing of the human beta-globin gene family during ontogeny reside proximally to the genes and immediately 5' to the human gamma- and beta globin genes. PMID- 12629214 TI - Triglyceride accumulation protects against fatty acid-induced lipotoxicity. AB - Excess lipid accumulation in non-adipose tissues is associated with insulin resistance, pancreatic beta-cell apoptosis and heart failure. Here, we demonstrate in cultured cells that the relative toxicity of two common dietary long chain fatty acids is related to channeling of these lipids to distinct cellular metabolic fates. Oleic acid supplementation leads to triglyceride accumulation and is well tolerated, whereas excess palmitic acid is poorly incorporated into triglyceride and causes apoptosis. Unsaturated fatty acids rescue palmitate-induced apoptosis by channeling palmitate into triglyceride pools and away from pathways leading to apoptosis. Moreover, in the setting of impaired triglyceride synthesis, oleate induces lipotoxicity. Our findings support a model of cellular lipid metabolism in which unsaturated fatty acids serve a protective function against lipotoxicity though promotion of triglyceride accumulation. PMID- 12629215 TI - Extending the definition of entropy to nonequilibrium steady states. AB - We study the nonequilibrium statistical mechanics of a finite classical system subjected to nongradient forces xi and maintained at fixed kinetic energy (Hoover Evans isokinetic thermostat). We assume that the microscopic dynamics is sufficiently chaotic (Gallavotti-Cohen chaotic hypothesis) and that there is a natural nonequilibrium steady-state rho(xi). When xi is replaced by xi + deltaxi, one can compute the change deltarho of rho(xi) (linear response) and define an entropy change deltaS based on energy considerations. When xi is varied around a loop, the total change of S need not vanish: Outside of equilibrium the entropy has curvature. However, at equilibrium (i.e., if xi is a gradient) we show that the curvature is zero, and that the entropy S(xi + deltaxi) near equilibrium is well defined to second order in deltaxi. PMID- 12629216 TI - Close identity of a pressure-stabilized intermediate with a kinetic intermediate in protein folding. AB - Atomic detailed structural study of a transiently existing folding intermediate is severely limited because of its short life. In ubiquitin, we found that a pressure-stabilized equilibrium conformer shares a common structural feature with the proline-trapped kinetic intermediate found in a pulse-labeling (1)H(2)H exchange NMR study [Briggs, M. S. & Roder, H. (1992) Proc. Natl. Acad. Sci. USA 89, 2017-2021]. The conformer is locally unfolded in the entire segment from residues 33 to 42 and in C-terminal residues 70-76. The close structural identity of an equilibrium intermediate stabilized under pressure with a transiently observed folding intermediate is likely to be general in terms of a folding funnel common to both experiments. PMID- 12629218 TI - Prospective identification of tumorigenic breast cancer cells. AB - Breast cancer is the most common malignancy in United States women, accounting for >40,000 deaths each year. These breast tumors are comprised of phenotypically diverse populations of breast cancer cells. Using a model in which human breast cancer cells were grown in immunocompromised mice, we found that only a minority of breast cancer cells had the ability to form new tumors. We were able to distinguish the tumorigenic (tumor initiating) from the nontumorigenic cancer cells based on cell surface marker expression. We prospectively identified and isolated the tumorigenic cells as CD44(+)CD24(-/low)Lineage(-) in eight of nine patients. As few as 100 cells with this phenotype were able to form tumors in mice, whereas tens of thousands of cells with alternate phenotypes failed to form tumors. The tumorigenic subpopulation could be serially passaged: each time cells within this population generated new tumors containing additional CD44(+)CD24( /low)Lineage(-) tumorigenic cells as well as the phenotypically diverse mixed populations of nontumorigenic cells present in the initial tumor. The ability to prospectively identify tumorigenic cancer cells will facilitate the elucidation of pathways that regulate their growth and survival. Furthermore, because these cells drive tumor development, strategies designed to target this population may lead to more effective therapies. PMID- 12629217 TI - The structure of the inter-SH2 domain of class IA phosphoinositide 3-kinase determined by site-directed spin labeling EPR and homology modeling. AB - Phosphoinositide (PI) 3-kinases catalyze the phosphorylation of the D3 position of the inositol ring of PI, and its phosphorylated derivatives and play important roles in many intracellular signal transducing pathways. Class IA PI3-kinases contain distinct regulatory (p85) and catalytic (p110) subunits. p110 is stabilized and inhibited by constitutive association with p85, and is disinhibited when the SH2 domains of p85 bind to tyrosyl-phosphorylated proteins. Because the two subunits do not dissociate, disinhibition of p110 presumably occurs by an allosteric mechanism. To explore the means by which p85 regulates the activity of p110, structures of the inter-SH2 domain of p85 were determined with and without phosphopeptide by using a combination of site directed spin labeling EPR and homology modeling and molecular dynamics. The inter-SH2 domain is assigned as a rigid anti-parallel coiled-coil whose primary function is to bind p110, facilitating inhibition of p110 by the N-terminal SH2 domain of p85. PMID- 12629219 TI - Essential function of alpha-calcium/calmodulin-dependent protein kinase II in neurotransmitter release at a glutamatergic central synapse. AB - A significant fraction of the total calciumcalmodulin-dependent protein kinase II (CaMKII) activity in neurons is associated with synaptic connections and is present in nerve terminals, thus suggesting a role for CaMKII in neurotransmitter release. To determine whether CaMKII regulates neurotransmitter release, we generated and analyzed knockout mice in which the dominant alpha-isoform of CaMKII was specifically deleted from the presynaptic side of the CA3-CA1 hippocampal synapse. Conditional CA3 alpha-CaMKII knockout mice exhibited an unchanged basal probability of neurotransmitter release at CA3-CA1 synapses but showed a significant enhancement in the activity-dependent increase in probability of release during repetitive presynaptic stimulation, as was shown with the analysis of unitary synaptic currents. These data indicate that alpha CaMKII serves as a negative activity-dependent regulator of neurotransmitter release at hippocampal synapses and maintains synapses in an optimal range of release probabilities necessary for normal synaptic operation. PMID- 12629220 TI - High-throughput approach for detection of DNA bending and flexibility based on cyclization. AB - We have developed a high-throughput approach to the labor-intensive problems of DNA cyclization, which we use to characterize DNA curvature and mechanical properties. The method includes a combinatorial approach to make the DNA constructs needed and automated real-time measurement of the kinetics using fluorescence. We validated the approach and investigated the flexibility of two kinds of nicked DNA and AT dinucleotide repeats. We found that, although the nicks hardly alter the bending flexibility, they significantly increase the torsional flexibility, and that the AT repeat has 28% (+/-12%) lower bending rigidity than a generic DNA sequence. PMID- 12629221 TI - Antigen presentation by keratinocytes directs autoimmune skin disease. AB - The antigen-presenting cells that initiate and maintain MHC class II-associated organ-specific autoimmune diseases are poorly defined. We now describe a new T cell antigen receptor (TCR) transgenic (Tg) model of inflammatory skin disease in which keratinocytes activate and are the primary target of autoreactive CD4(+) T cells. We previously generated keratin 14 (K14)-A(beta)b mice expressing MHC class II only on thymic cortical epithelium. CD4(+) T cells from K14-A(beta)b mice fail to undergo negative selection and thus have significant autoreactivity. The TCR genes from an autoreactive K14-A(beta)b CD4 hybridoma were cloned to produce a TCR Tg mouse, 2-2-3. 2-2-3 TCR Tg cells are negatively selected in WT C57BL6 mice but not in 2-2-3K14-A(beta)b mice. Interestingly, a significant number of mice that express both the K14-A(beta)b transgene and the autoreactive 2-2-3 TCR spontaneously develop inflammatory skin disease with mononuclear infiltrates, induction of MHC class II expression on keratinocytes, and T helper 1 cytokines. Disease can be induced by skin inflammation but not solely by activation of T cells. Thus, cutaneous immunopathology can be directed through antigen presentation by tissue-resident keratinocytes to autoreactive TCR Tg CD4(+) cells. PMID- 12629222 TI - The matrix attachment region in the Chinese hamster dihydrofolate reductase origin of replication may be required for local chromatid separation. AB - Centered in the Chinese hamster dihydrofolate reductase origin of replication is a prominent nuclear matrix attachment region (MAR). Indirect lines of evidence suggested that this MAR might be required for origin activation in early S phase. To test this possibility, we have deleted the MAR from a Chinese hamster ovary variant harboring a single copy of the dihydrofolate reductase locus. However, 2D gel replicon mapping shows that removal of the MAR has no significant effect either on the frequency or timing of initiation in this locus. Rather, fluorescence in situ hybridization studies on cells swollen under either neutral or alkaline conditions show that deletion of the MAR interferes with local separation of daughter chromatids. This surprising result provides direct genetic evidence that at least a subset of MARs performs an important biological function, possibly related to chromatid cohesion and separation. PMID- 12629223 TI - Control of neurotransmitter release by an internal gel matrix in synaptic vesicles. AB - Neurotransmitters are stored in synaptic vesicles, where they have been assumed to be in free solution. Here we report that in Torpedo synaptic vesicles, only 5% of the total acetylcholine (ACh) or ATP content is free, and that the rest is adsorbed to an intravesicular proteoglycan matrix. This matrix, which controls ACh and ATP release by an ion-exchange mechanism, behaves like a smart gel. That is, it releases neurotransmitter and changes its volume when challenged with small ionic concentration change. Immunodetection analysis revealed that the synaptic vesicle proteoglycan SV2 is the core of the intravesicular matrix and is responsible for immobilization and release of ACh and ATP. We suggest that in the early steps of vesicle fusion, this internal matrix regulates the availability of free diffusible ACh and ATP, and thus serves to modulate the quantity of transmitter released. PMID- 12629224 TI - Regulated subset of G1 growth-control genes in response to derepression by the Wnt pathway. AB - Pitx2 is a bicoid-related homeodomain factor that is required for effective cell type-specific proliferation directly activating a specific growth-regulating gene cyclin D2. Here, we report that Pitx2, in response to the Wntbeta-catenin pathway and growth signals, also can regulate c-Myc and cyclin D1. Investigation of molecular mechanisms required for Pitx2-dependent proliferation, in these cases, further supports a nuclear role for beta-catenin in preventing the histone deacetylase 1-dependent inhibitory functions of several DNA-binding transcriptional repressors, potentially including E2F4p130 pocket protein inhibitory complex, as well as lymphoid enhancer factor 1 and Pitx2, by dismissal of histone deacetylase 1 and loss of its enzymatic activity. Thus, beta-catenin plays a signal-integrating role in Wnt- and growth factor-dependent proliferation events in mammalian development by both derepressing several classes of repressors and by activating Pitx2, regulating the activity of several growth control genes. PMID- 12629226 TI - Does serotonin have trophic effects in temporal lobe epilepsy? PMID- 12629227 TI - Development in pain and neurologic complaints after whiplash: a 1-year prospective study. AB - OBJECTIVE: To prospectively examine the course of pain and other neurologic complaints in patients with acute whiplash injury and in controls with acute ankle injury. METHODS: Patients with acute whiplash (n = 141) and ankle-injured controls (n = 40) were consecutively sampled, and underwent interview and examination after 1 week and 1, 3, 6, and 12 months. Outcome measures were pain intensity, pain frequency, and associated symptoms. RESULTS: Initial overall pain intensity above lower extremities (pain in neck, head, shoulder-arm, and low back) was similar in patients with whiplash (median Visual Analogue Scale [VAS](0 100) of 20 [25th and 75th percentile, 4, 39]) and ankle-injured controls (median VAS(0-100) of 15 [5, 34]). Whiplash-injured patients reported median overall VAS(0-100) pain intensity above lower extremities of 23 (12, 40) after 11 days and 14 (12, 40) after 1 year. Controls reported pain intensity of 0 (0, 4) after 12 days and 0 (0, 9) after 1 year. Reported overall pain frequency above lower extremities was 96% after 11 days and 74% after 1 year in whiplash-injured patients and 33% after 12 days and 47% after 1 year in controls. Associated neurologic symptoms were two to three times more common after whiplash injury. Correlation was found between pain intensity and associated symptoms in whiplash injured patients but not controls. CONCLUSION: Pain occurs with high frequency but low intensity after whiplash and ankle injury. Associated neurologic symptoms were not correlated to pain in ankle-injured controls, but were correlated to pain in patients with whiplash injury. Persistent symptoms in whiplash-injured patients may be caused by both specific neck injury-related factors and nonspecific post-traumatic reactions. Disability was only encountered in the whiplash group. PMID- 12629228 TI - PET imaging of 5-HT1A receptor binding in patients with temporal lobe epilepsy. AB - BACKGROUND: Activation of central serotonin (5-HT)1A receptors, found in high density in brainstem raphe, hippocampus, and temporal neocortex, exerts an anticonvulsant effect in various experimental seizure models. To test the hypothesis that 5-HT1A receptor binding is reduced in human epileptic foci, PET imaging was performed using the radioligand [18F]trans-4-fluoro-N-2-[4-(2 methoxyphenyl)piperazin-1-yl]ethyl]-N-(2-pyridyl)cyclohexanecarboxamide ([18F]FCWAY), a selective 5-HT1A receptor antagonist, in patients with temporal lobe epilepsy and normal controls. METHODS: MRI and PET were performed using [15O]water and [18F]FCWAY in 10 controls and in 12 patients with temporal lobe epilepsy confirmed on ictal video-EEG; patients also underwent [18F]fluorodeoxyglucose PET. Using quantitative PET image analysis, regional values were obtained for [18F]FCWAY volume of distribution (V), cerebral blood flow (CBF), and glucose cerebral metabolic rate (CMRglc). Hippocampal volume (HV) was also measured with MRI. [18F]FCWAY V PET and MR measures were compared within patients and controls using paired t-tests; grouped comparisons were made with two sample t-tests. RESULTS: Lower [18F]FCWAY V was found ipsilateral than contralateral to the epileptic focus in inferior medial (IMT) and lateral (ILT) temporal regions of patients (ILT 47.4 +/- 6.1 vs 61.8 +/- 6.1, p < 0.01; IMT 52 +/- 4.6 vs 67.0 +/- 6.0, p < 0.01). [18F]FCWAY V was 29% lower in raphe and 34% lower in the ipsilateral thalamic region of patients than controls. In ILT, mean [18F]FCWAY V asymmetry index (AI) was significantly greater than mean CBF and mean CMRglc AI. Mean [18F]FCWAY V AI in IMT was greater than mean HV AI, but the difference was not significant. CONCLUSION: These findings support the hypothesis of reduced serotonin receptor binding in temporal lobe epileptic foci. PMID- 12629225 TI - The impact of parietal cells on Helicobacter pylori tropism and host pathology: an analysis using gnotobiotic normal and transgenic mice. AB - Helicobacter pylori infection of the human stomach is common and typically benign, although a subset of hosts develops severe pathology. Infection occurs in an organ with distinct microenvironments characterized by pronounced differences in the composition of acid-producing parietal cells. In this study, we examine determinants of bacterial tropism to various gastric niches by using germ-free normal and transgenic mice with an engineered parietal cell ablation. Mice were colonized for 8 weeks with a clinical isolate (Hp1) that expresses adhesins recognized by epithelial NeuAcalpha2,3Galbeta1,4 glycan receptors. In normal mice, Hp1 has tropism for a parietal cell-deficient niche where sialylated glycans are expressed by a narrow band of pit cells positioned at the boundary between the squamous epithelium (forestomach) and the proximal glandular epithelium. Lymphoid aggregates that develop in this niche, but not elsewhere in the stomach, were analyzed by GeneChip and quantitative RT-PCR studies of laser capture microdissected mucosa and yielded a series of biomarkers indicative of immune cell activation and maturation. Genetic ablation of parietal cells produced a new source of NeuAcalpha2,3Galbeta1,4 glycans in amplified gastric epithelial lineage progenitors, with accompanying expansion of Hp1 within the glandular epithelium. Lymphoid aggregates that develop in this formerly acid protected epithelium have molecular features similar to those observed at the forestomach/glandular junction. These findings demonstrate the important roles played by parietal cells and glycan receptors in determining the positioning of H. pylori within the gastric ecosystem, and emphasize the need to consider the evolution of pathology within a given host in a niche-specific context. PMID- 12629229 TI - Alpha-[11C] methyl-L-tryptophan and glucose metabolism in patients with temporal lobe epilepsy. AB - OBJECTIVE: To determine whether metabolism in the brain serotonergic system, including the kynurenine pathway, is involved in temporal lobe epilepsy (TLE). METHODS: The authors studied 14 patients with intractable TLE by PET using alpha [11C] methyl-L-tryptophan (alpha-MTrp) and 2-[18F]-fluoro-deoxy-glucose (FDG) and compared results with 21 healthy control subjects who had alpha-MTrp PET studies. Seven patients had unilateral hippocampal atrophy (HA), and seven had normal hippocampal volumes (NV). The regional uptake constant (K*) for alpha-MTrp and regional FDG uptake were calculated in regions with high serotonergic innervation, including the hippocampus, amygdala, lateral temporal lobe, frontal lobe, thalamus, lenticular nucleus, and cingulate cortex. RESULTS: A significant increase of alpha-MTrp uptake was observed in the hippocampus ipsilateral to the seizure focus in seven TLE patients with NV compared to seven patients with HA as well as to healthy controls. In patients with TLE, glucose utilization in the lateral temporal lobe ipsilateral to the seizure focus was correlated negatively with K* for alpha-MTrp in the ipsilateral hippocampus and positively with K* in the ipsilateral lenticular nucleus and cingulate cortex. Glucose utilization in the frontal lobe ipsilateral to the seizure shows a reduction in the glucose utilization which relates to the increase in the alpha-MTrp uptake in the ipsilateral lateral temporal lobe. CONCLUSION: This study demonstrates dysfunction of the serotonergic system, which could include metabolism through the kynurenine pathway in TLE patients with normal hippocampal volumes. alpha MTrp PET studies might be useful for lateralizing the epileptic focus in TLE patients with normal hippocampal volumes. PMID- 12629230 TI - Generalized epilepsy in hypothalamic hamartoma: evolution and postoperative resolution. AB - OBJECTIVE: To better understand the epileptogenesis of symptomatic generalized epilepsy in patients with hypothalamic hamartoma and intractable epilepsy, many of whom experience remission of generalized seizures and slow spike-wave discharges following surgery. METHODS: The authors documented the evolution of symptomatic generalized epilepsy in 12 of 20 children who underwent transcallosal microsurgical hypothalamic hamartoma resection. In seven patients they recorded intraoperative EEG from the hamartoma and simultaneously from the scalp and frontal cortex before, during, and after resection. RESULTS: Gelastic seizures began on average at 6 months of age (range birth to 3 years); tonic seizures began at 6 years (range 2 months to 9 years). Normal EEG were reported in early childhood; thereafter, abnormalities were progressive. Interictal spike-wave was recorded intraoperatively over the scalp and cortex in six patients, but not from the hypothalamic hamartoma. Hamartoma resection had no immediate effect on cortical spike-wave, but waking spike-wave was absent in seven patients on subsequent postoperative EEG. Tonic seizures ceased in 11 of 12 patients, but 6 of these had postoperative generalized seizures that resolved over 1 to 6 months. CONCLUSION: Gelastic seizures in hypothalamic hamartoma arise from the hamartoma itself; the interictal spike-wave does not. The evolution of EEG abnormalities, the development of generalized seizures years after onset of gelastic seizures, and the postoperative running down of interictal spike-wave and generalized seizures in these patients may reflect secondary epileptogenesis. PMID- 12629231 TI - Modulation of motor cortex excitability by pallidal stimulation in patients with severe dystonia. AB - OBJECTIVE: To study the influence of continuous high-frequency electrical stimulation with electrodes implanted in the globus pallidus internus (GPi) on motor cortex excitability in nine patients with dystonia. METHODS: Short-term effects related to switching off and on the deep brain stimulator were investigated >3 months after electrode implantation. Activation of motor cortical excitatory and inhibitory neurons was examined with transcranial magnetic stimulation (TMS) by analysis of electromyographic activity in the hand muscles. Parameters of corticospinally mediated excitatory motor responses included latency, threshold, and response sizes with increasing stimulus intensities (stimulus-response curves). Other measures of motor cortex excitability comprised the duration of the contralateral silent period and intracortical inhibition and facilitation in a paired-pulse paradigm. RESULTS: Switching off GPi stimulation led to a decrease of motor cortex excitability, as reflected by an increase in motor thresholds (GPi stimulation on 37.5 +/- 6.1%, mean +/- SD; GPi stimulation off for 15 to 120 minutes, 40.5 +/- 6.7% of maximum stimulator output), and reduced the size of contralateral responses in the stimulus-response curves established for relaxed muscles. The changes were reversible within minutes after switching on GPi stimulation. They were associated with mild changes of dystonia. By contrast, measures of intracortical inhibition were not altered by switching off GPi stimulation. Spinal excitability did not change as assessed by H-reflex. CONCLUSION: GPi stimulation influences motor cortex excitability by a rapid modulation of thalamocortical outputs. PMID- 12629232 TI - Grammatical and resource components of sentence processing in Parkinson's disease: an fMRI study. AB - BACKGROUND: Sentence comprehension requires linguistic processing as well as cognitive resources such as working memory (WM) and information-processing speed (IPS). The authors hypothesize that sentence comprehension difficulty in patients with mild PD is due to degradation of the large-scale neural network that supports cognitive resources during sentence processing. OBJECTIVE: To understand the neural basis for sentence comprehension difficulty in PD. METHOD: Regional brain activity with blood oxygenation level-dependent fMRI was monitored while seven PD patients and nine healthy seniors answered a simple probe about written sentences that vary in their grammatical and cognitive resource properties. RESULTS: Healthy seniors recruited posterolateral temporal and ventral inferior frontal regions of the left hemisphere, brain regions associated with grammatical processing that were also activated by PD patients. Healthy seniors also recruited left dorsal inferior frontal, right posterolateral temporal, and striatal regions that are associated with cognitive resources during sentence processing. Direct contrasts showed that striatal, anteromedial prefrontal, and right temporal regions are recruited to a significantly lesser degree in PD, but these patients have increased activation of right inferior frontal and left posterolateral temporal-parietal areas during sentence comprehension. CONCLUSION: These findings associate impaired sentence comprehension in PD with interruption of a large-scale network important for cognitive resources during sentence processing. These results also imply compensatory up-regulation of cortical activity that allows patients with mild PD to maintain sentence comprehension accuracy. PMID- 12629233 TI - Temporal discrimination of cross-modal and unimodal stimuli in generalized dystonia. AB - BACKGROUND: Motor and nonmotor timing functions and cross-modal processing of visual-tactile signals may be linked to basal ganglia. These neural structures are thought to be dysfunctional in dystonia. OBJECTIVE: To test whether cross modal stimulation influences deficits of temporal discrimination in dystonia. METHODS: Eight patients with generalized dystonia and 10 control subjects were asked to discriminate whether pairs of unimodal (tactile or visual) and cross modal (visual and tactile) stimuli were simultaneous or sequential and, in the latter case, which stimulus preceded the other. Visual stimuli consisted of red lights and tactile stimuli of non-noxious electrical shocks. Intervals between stimuli in each pair were increased from 0 to 400 msec (in steps of 10 msec). RESULTS: Patients with dystonia recognized the asynchrony between the experimental stimuli and judged correctly which stimulus in a pair came first, at significantly longer intervals than did controls. Moreover, differences in performance between patients and controls were maximal for cross-modal stimuli. The defective performance of patients with dystonia in the cross-modal combinations showed a high positive correlation with the severity of symptoms. CONCLUSION: Patients with generalized dystonia present with difficulties both in timing functions and in cross-modal processing of visual-tactile stimuli. PMID- 12629234 TI - Tremor response to polarity, voltage, pulsewidth and frequency of thalamic stimulation. AB - BACKGROUND: Thalamic deep brain stimulation ameliorates essential and parkinsonian tremors refractory to medications. Stimulus voltage, polarity configuration, frequency, and pulsewidth can each be adjusted in order to optimize tremor control and maximize battery life. The relative impacts of these programmable variables have not previously been quantified. METHODS: The thalamus of 11 patients (bilaterally in 2) was studied 4 to 59 months postoperatively. The stimulator was inactivated and medications withheld for 12 hours, and optimal electrode contacts were selected. Stimulation followed at a range of voltages (0, 1, 2, 3, or 4 V), pulsewidths (60, 90, or 120 micros), and frequencies (130, 160, or 185 Hz) for both monopolar and bipolar configurations. Seventy-eight combinations of variables were programmed in random sequence. Postural and action tremors were measured with an electromagnetic tracker, tremor was subjectively graded, and side effects were noted. RESULTS: Voltage was consistently predictive of tremor response. Mean postural tremor amplitude in PD fell from 6.4 cm at 0 V to 2.6, 1.0, 0.3, and 0 cm at 1 through 4 V (bipolar configuration). The voltage response curve for essential tremor was flatter. The monopolar configuration was 10 to 25% more effective than bipolar. The longest pulsewidth tested was up to 30% more effective than the shortest, but frequency changes had little effect on tremor amplitude. Side effects occurred only with monopolar stimulation, and the only setting that was intolerable for the majority was 4 V, 120 micros, and 185 Hz. CONCLUSION: Bipolar deep brain stimulation at 90 micros, 130 Hz, adjusting the voltage up to 3 V, tends to be effective and well tolerated. Monopolar provides similar benefits for lower voltage, but side effects become common at 3 or 4 V. PMID- 12629235 TI - Caffeine, postmenopausal estrogen, and risk of Parkinson's disease. AB - BACKGROUND: Men who regularly consume caffeinated drinks have a lower risk of PD than do nondrinkers, but this relation has not been found in women. Because this sex difference could be due to hormonal effects, the authors examined prospectively the risk of PD according to use of postmenopausal hormones and caffeine intake among participants in the Nurses' Health Study. METHODS: The study population comprised 77,713 women free of PD, stroke, or cancer at baseline, who were postmenopausal at baseline or reached menopause before the end of the study. During 18 years of follow-up the authors documented 154 cases of PD. RESULTS: Overall, the risk of PD was similar in women using hormones and women who never used hormones (relative risk 1.02, 95% CI 0.69 to 1.52). Use of hormones, however, was associated with a reduced risk of PD among women with low caffeine consumption (RR 0.39, 95% CI 0.13 to 1.17), and with increased risk among women with high caffeine consumption (RR 2.44, 95% CI 0.75 to 7.86; p for interaction = 0.01). Among hormone users, women consuming six or more cups of coffee per day had a fourfold higher risk of PD (RR 3.92, 95% CI 1.49 to 10.34; p = 0.006) than did women who never drink coffee. CONCLUSION: These results suggest that caffeine reduces the risk of PD among women who do not use postmenopausal hormones, but increases risk among hormone users. Clinical trials of caffeine or estrogens in women should avoid the combined use of these agents. PMID- 12629236 TI - Heterozygosity for a mutation in the parkin gene leads to later onset Parkinson disease. AB - BACKGROUND: The vast majority of the parkin mutations previously identified have been found in individuals with juvenile or early onset PD. Previous screening of later onset PD cohorts has not identified substantial numbers of parkin mutations. METHODS: Families with at least two siblings with PD were ascertained to identify genes contributing to PD susceptibility. Screening of the parkin gene, by both quantitative PCR and exon sequencing, was performed in those families with either early onset PD (age onset < or =50 years) or positive lod score with a marker in intron 7 of the parkin gene. RESULTS: A total of 25 different mutations in the parkin gene were identified in 103 individuals from 47 families. Mutations were found in both parkin alleles in 41 of the individuals, whereas a single mutation in only one of the two parkin alleles was observed in 62 individuals. Thirty-five of the subjects (34%) with a parkin mutation had an age at onset of 60 years or above with 30 of these 35 (86%) having a detectable mutation on only one parkin allele. Few significant clinical differences were observed among the individuals with two, one, or no mutated copies of the parkin gene. CONCLUSION: Mutations in the parkin gene occur among individuals with PD with an older age at onset (> or =60 years) who have a positive family history of the disease. In addition, the clinical findings of parkin-positive individuals are remarkably similar to those without mutations. PMID- 12629237 TI - A visuospatial variant of mild cognitive impairment: getting lost between aging and AD. AB - BACKGROUND: AD causes visuospatial disorientation that is associated with posterior cortical atrophy and impaired visual motion processing. OBJECTIVE: The authors characterized memory capacity and visual motion processing in young normal (YN) and older normal (ON) adult subjects and in patients with mild cognitive impairment (MCI) and AD to see if deficits in these realms occur as isolated impairments. METHODS: Each participant underwent neuropsychological testing and gave push-button responses to indicate perception of panoramic visual motion stimuli. RESULTS: One fifth of the ON subjects, one third of the patients with MCI, and half of the patients with AD showed increasingly pervasive impairments of visual motion perception. These impairments were associated with poorer performance on the Money Road Map test of spatial navigation but not with verbal or visual memory deficits. CONCLUSION: Impaired visual motion processing may accompany memory deficits in MCI or AD, or may occur alone in otherwise intact ON subjects. This suggests that visuospatial impairment may develop as an independent sign of neurodegenerative disease, possibly preceding the clinical onset of AD. PMID- 12629238 TI - Detection of genomic viral RNA in nerve and muscle of patients with HCV neuropathy. AB - BACKGROUND: Hepatitis C virus (HCV)-associated neuropathy is usually associated with mixed cryoglobulinemia (MC) and vasculitis. MC may contain viral RNA, and tissues showing vasculitis may contain intracellular HCV. Local HCV replication remains to be evidenced. OBJECTIVE: To delineate the spectrum of HCV-associated neuropathy and to assess the presence of HCV in nerve and muscle tissues. METHODS: Thirty consecutive HCV-infected patients with peripheral neuropathy were included. Genomic and replicative strands of HCV RNA were detected in both nerve and muscle biopsy samples using distinctive reverse transcription nested PCR. RESULTS: Neuropathy was consistent with distal axonal polyneuropathy (DPN) in 25 of 30 patients, mononeuropathy multiplex (MM) in 3 of 30, and demyelinating polyneuropathy in 2 of 30. Pain was present in 18 of 30 patients and MC in 16 of 30. Biopsy showed inflammatory vascular lesions in 26 of 30 patients (87%), including necrotizing arteritis (6/30), small-vessel vasculitis (12/30) of either the lymphocytic (9/12) or the leukocytoclastic (3/12) type, and perivascular inflammatory infiltrates (8/30). All patients with necrotizing arteritis had DPN and positive MC detection. Both pain (p < 0.03) and positive MC detection (p < 0.01) were associated with the presence of vasculitis. Positive-strand genomic HCV RNA was detected in tissues of 10 of 30 patients (muscle 9, nerve 3). In contrast, negative-strand replicative RNA was never detected. Genomic RNA was found in nerve tissue samples showing vasculitis (necrotizing arteritis 2, small vessel lymphocytic vasculitis 1). CONCLUSION: Painful DPN associated with MC and neuromuscular vasculitis is the most frequent type of HCV neuropathy. The usual detection of MC and the lack of local HCV replication indicate that HCV neuropathy results from virus-triggered immune-mediated mechanisms rather than direct nerve infection and in situ replication. PMID- 12629239 TI - Prognosis in amyotrophic lateral sclerosis: a population-based study. AB - BACKGROUND: Accurate information on prognosis of ALS is useful to patients, families, and clinicians. METHODS: In a population-based study of ALS in western Washington, the authors assembled a cohort of 180 patients with incident ALS between 1990 and 1994. Information on potential prognostic factors was collected during an in-person interview. Patients also completed the Medical Outcomes Study Short Form 36 (SF-36). Vital status through December 1999 was known for all patients. RESULTS: Median survival was 32 months from onset of symptoms and 19 months from diagnosis. The 5-year survival after diagnosis was 7%. Older age and female sex were strongly associated with poor survival. In multivariable Cox proportional hazards regression models, factors significantly and independently associated with a worse prognosis included older age, any bulbar features at onset, shorter time from symptom onset to diagnosis, lack of a marital partner, and residence in King County. Recursive partitioning identified age, time from symptom onset to diagnosis, and marital status as the strongest predictors of survival. Good summary scores for physical health on the SF-36, but not for mental health, were significantly associated with longer survival than poor scores. CONCLUSION: These findings are consistent with other population-based studies of ALS and confirm its pernicious nature. Older age, female sex, any bulbar features at onset, short time from symptom onset to diagnosis, lack of a marital partner, and disease severity are key prognostic factors. Serial measurement of severity would likely improve predictions. PMID- 12629240 TI - Proton MRS profile of cerebral metabolic abnormalities in Krabbe disease. AB - BACKGROUND: Krabbe disease (globoid cell leukodystrophy [GLD]) is an autosomal recessive lysosomal disorder affecting the central and peripheral nervous system. The authors performed MRS to characterize metabolic alterations and their regional variation in brain tissue in GLD in vivo. METHODS: Abnormalities of cerebral metabolite concentrations were assessed in seven patients with biochemically proven GLD-four with infantile, two with juvenile, and one with adult subtype-using quantitative localized proton MRS of standardized brain regions. RESULTS: In infantile GLD, pronounced elevation of both myo-inositol and choline-containing compounds in affected white matter reflected demyelination and glial proliferation. The accompanying decrease of N-acetylaspartate pointed to neuroaxonal loss. Gray matter showed similar, albeit much milder alterations. In juvenile GLD, MRS indicated astrocytosis with minor neuroaxonal damage in white matter. In a patient with adult GLD, results of MRS of affected white matter were close to normal. MRS data are in agreement with histopathologic features of GLD. CONCLUSION: Proton MRS provides a powerful tool for assessing metabolic disturbances and the extent of brain damage noninvasively in GLD. PMID- 12629241 TI - Interobserver variability in the radiological assessment of response to chemotherapy in glioma. AB - OBJECTIVE: To assess the interobserver variability in the radiologic assessment of response to chemotherapy in patients with recurrent glioma. METHODS: Five clinicians with experience in the treatment and follow-up of patients with glioma measured tumor size in 20 pairs of CT and 20 pairs of MRI scans of 35 patients who had been treated with chemotherapy for recurrent glioma. Tumor size was defined as the product of the two largest perpendicular enhancing tumor diameters on the postcontrast images. To assess the interobserver variability in the measurements of tumor size, and in the classification according to the widely used Macdonald response criteria, intraclass correlation coefficients (ICC) and weighted kappa values were calculated. RESULTS: Substantial interobserver agreement was noted in the manual, two-dimensional measurements of tumor size on CT and MRI in patients treated with chemotherapy for recurrent glioma (overall ICC 0.64). Classification of response to chemotherapy according to the Macdonald criteria resulted in moderate interobserver agreement (overall weighted kappa 0.51). In 65% of evaluated CT and in 55% of evaluated MRI studies, no complete consensus was found for the categorical tumor response measurement. CONCLUSION: The radiologic assessment of response to chemotherapy in patients with recurrent glioma is susceptible to considerable interobserver variability. This underlines the difficulties that arise in scoring response to chemotherapy by conventional radiologic techniques. PMID- 12629242 TI - Influence of education on the relationship between white matter lesions and cognition. AB - OBJECTIVE: To test the hypothesis that education level modulates the effects of cerebral white matter hyperintensities (WMH) on cognition in a large population based study. METHODS: A total of 845 elderly subjects aged 64 to 76 years who enrolled in a longitudinal study on cognitive decline and vascular aging had an MRI examination. Cognitive functions were assessed by Mini-Mental State Examination, Trail Making Test Part B, Digit Symbol Substitution Test of the Wechsler Adult Intelligence Scale-Revised, Finger Tapping Test, Word Fluency Test, and Raven Progressive Matrix. MRI scans were interpreted visually using a standardized scale for rating WMH. RESULTS: Severe WMH were present in 17% of the participants who had lower performances on tests involving attention tasks. In participants with a lower level of education, presence of severe WMH was significantly associated with lower cognitive performances. This was found for all cognitive tests. Conversely, in participants with a high level of education, there was no significant association between severity of WMH and level of cognitive functions. CONCLUSION: Education modulates the consequences of WMH on cognition. Participants with a high level of education were protected against the cognitive deterioration related to vascular insults of the brain. PMID- 12629243 TI - Risk factors for fever in the neurologic intensive care unit. AB - OBJECTIVE: To identify risk factors for fever among patients treated in a neurologic intensive care unit (NICU). METHODS: The authors prospectively studied the frequency and causes of fever, defined as a patient's first temperature > or =101 degrees F (38.3 degrees C), among 387 patients consecutively admitted to their NICU. After identifying risk factors for 1) any fever, 2) infectious fever, and 3) unexplained fever using logistic regression, they calculated disease specific adjusted odds ratios for developing these types of fever among 12 diagnostic groups. RESULTS: Fever occurred in 23% (87/387) of patients. Fifty-two percent of fevers were explained by infection (predominantly pneumonia or bronchitis), and 28% were unexplained despite a complete diagnostic evaluation. NICU length of stay was a risk factor for all three types of fever (all p < 0.004); other risk factors included depressed level of consciousness for any fever (p = 0.005) and infectious fever (p = 0.048), endotracheal intubation for infectious fever (p = 0.01), and intraventricular catheterization for unexplained fever (p = 0.004). Subarachnoid hemorrhage increased the risk of both infectious and unexplained fever, even after adjusting for these risk factors (p = 0.006). CONCLUSION: Fever occurs in nearly 25% of NICU patients, and is associated with increased length of stay and depressed level of consciousness. Endotracheal intubation is a risk factor for infectious fever, whereas intraventricular catheterization is a risk factor for unexplained fever, which suggests a role for ventricular hemorrhage in the pathogenesis of "central" fever. Subarachnoid hemorrhage increases the risk of developing fever of all types. PMID- 12629244 TI - Prospective evaluation of neurological complications after allogeneic bone marrow transplantation. AB - OBJECTIVE: To determine the spectrum and frequency of neurologic sequelae after allogeneic bone marrow transplantation (BMT) and to define a risk profile of the patients. METHODS: A prospective follow-up of 71 allogeneic bone marrow recipients 14 +/- 3 months after transplantation. Patients underwent a neurologic examination, a neuropsychological test battery, and cranial MRI before and after BMT. RESULTS: A large proportion of patients (65%) developed sequelae after BMT. Acute complications of defined etiology occurred in 18% of the patients and led to death in 9% of the study population. A total of 47% of the patients developed new neurologic abnormalities of undefined origin that were mild and subacute and predominantly affected the peripheral nervous system. The cognitive and neuroradiologic outcome was favorable in a majority of these patients, but a small subgroup exhibited cognitive deterioration and white matter lesions. Risk factor analysis identified acute graft-versus-host disease (GvHD) and other variables partly related to GvHD such as long-lasting immunosuppression as the main predictors of sequelae after allogeneic BMT. The authors have established an association with various factors but, owing to the observational character of this study, conclusions about the etiology of the findings are unclear. CONCLUSION: Neurologic complications significantly contribute to the morbidity and mortality of patients receiving allogeneic BMT. Subclinical abnormalities, cognitive deficits, and white matter lesions detected 1 year after BMT in a subgroup of patients may be related to more extensive CNS changes observed after transplantation in an earlier retrospective study and may be associated with the risk factor chronic GvHD/immunosuppression. PMID- 12629245 TI - Neuromyelitis optica: clinical predictors of a relapsing course and survival. AB - BACKGROUND: The relapsing form of neuromyelitis optica (NMO) is characterized by recurrent optic neuritis and myelitis, usually leading to severe, permanent, relapse-related neurologic impairment (e.g., blindness, paraplegia) within 5 years. Aggressive therapy aimed at relapse prevention initiated soon after disease onset may be expected to have a relatively greater impact on early relapse-related disability in NMO than in typical MS. Early prediction of a relapsing course and subsequent disease severity would facilitate design and implementation of clinical trials of such therapies. METHODS: A database of clinical and laboratory features of patients with NMO (n = 80) was used to develop potentially useful models predictive of a relapsing disease course and of subsequent disease severity as measured by survival. RESULTS: Predictors of a relapsing course were longer interattack interval between the first two clinical events (rate ratio [RR] = 2.16; per month increase), older age at onset (RR = 1.08; per year increase), female sex (RR = 10.0, female vs male), and less severe motor impairment with the sentinel myelitis event (RR = 0.48; per severity scale point increase). A history of other autoimmune disease (RR = 4.15; presence vs absence), higher attack frequency during the first 2 years of disease (RR = 1.21; per attack), and better motor recovery following the index myelitis event (RR = 1.84; per point increase) were associated with mortality due to relapsing NMO. CONCLUSIONS: These predictive models identify several clinical features, each available at diagnosis or early in the disease course, that predict relapsing disease and survival. These results may be useful to identify patients at high risk for severe, relapsing neuromyelitis optica in order to initiate early therapy for relapse prevention and to design clinical trials to study such interventions. PMID- 12629246 TI - The effect of interferon beta-1b on quantities derived from MT MRI in secondary progressive MS. AB - BACKGROUND: Magnetization transfer (MT) MRI can provide in vivo markers reflecting the severity of irreversible, MS-related brain damage occurring within and outside T2-visible lesions. OBJECTIVE: To assess the effect of interferon (IFN) beta-1b treatment on the accumulation of brain damage in patients with secondary progressive (SP) MS, measured using MT MRI. METHODS: Eighty-two patients with SPMS from five centers participating in a European, multicenter, double-blind, placebo-controlled trial of IFNbeta-1b in SPMS underwent brain T2 weighted and MT MRI at baseline. Evaluable follow-up data were available for 75 patients at 12 months, 54 at 24 months, and 47 at 36 months. MT MRI scans were postprocessed and analyzed to obtain histograms of MT ratio (MTR) values from the whole brain. A region of interest-based analysis of MTR values from the normal appearing white matter (NAWM) was also performed. RESULTS: In both the treatment arms, there was a decrease of average brain MTR values from baseline to month 24 (mean change -4.9%) and month 36 (mean change -4.3%). These changes were significant for the placebo group at both timepoints and for the IFNbeta-1b group at month 24 only, with no significant treatment effect. A decrease of NAWM MTR was also observed, with no significant difference between the two treatment arms. CONCLUSION: In this cohort of patients with secondary progressive MS, interferon beta-1b did not show an overall effect on the worsening of magnetization transfer MRI measures, when compared with placebo. The data show that change in magnetization transfer ratio is a promising tool for monitoring disease evolution in secondary progressive MS and that the information obtained from magnetization transfer MRI complements that obtained from MRI measures of lesion load and inflammation. PMID- 12629247 TI - Cutaneous T-cell lymphoma imitating gelsolin amyloidosis. PMID- 12629248 TI - Possible association of the tau H1/H1 genotype with primary progressive aphasia. AB - The authors screened for tau gene mutations and polymorphisms to determine whether genetic variation at or near the tau locus contributes to the development of primary progressive aphasia (PPA). No mutations were detected in 25 patients with PPA. However, a significant overrepresentation of the tau H1/H1 genotype, also found in progressive supranuclear palsy and corticobasal degeneration, was found in the PPA group. Whether tau haplotypes have a primary causal role or whether they affect the topology of neurodegeneration remains to be determined. PMID- 12629249 TI - Remarkable improvement in adult Leigh syndrome with partial cytochrome c oxidase deficiency. AB - Leigh syndrome (LS) is a heterogeneous disorder, usually due to a defect in oxidative metabolism. Typically, signs and symptoms commence in infancy or childhood, although rare cases of adult onset have been described. Progressive deterioration is the norm. The authors describe a 22-year-old woman with partial cytochrome c oxidase deficiency who developed fulminant LS following an acute febrile illness and who subsequently showed dramatic clinical and neuroradiologic improvement. PMID- 12629250 TI - Phenotypic variability of aprataxin gene mutations. AB - The clinical and genetic features of three non-Portuguese and non-Japanese patients with aprataxin gene mutations are reported. Patient 1 came from Italy and presented with typical ataxia with ocular motor apraxia (OMA). She was homozygous for the W279X nonsense mutation, which is associated with the Portuguese founding haplotype. Patients 2 and 3 were French siblings and did not present with either OMA or hypoalbuminemia. They were compound heterozygous for the nonsense W279X mutation and a missense K197Q mutation. PMID- 12629251 TI - Unilateral absence of the basal ganglia plus epilepsy without motor symptoms. AB - A patient with absence of the basal ganglia and refractory epilepsy without impairment of pyramidal or extrapyramidal motor function is reported. Imaging findings suggest a vascular insult as etiology. Preserved motor function could be explained by neuronal plasticity involving contralateral corticostriatal and pallidothalamic connections and points to a lesion received in early pregnancy. PMID- 12629252 TI - Peri-ictal water drinking lateralizes seizure onset to the nondominant temporal lobe. AB - The authors describe seven patients with medically refractory temporal lobe epilepsy whose seizures were associated with peri-ictal water drinking behavior. Presurgical evaluation, including video-EEG monitoring, MRI, SPECT, and neuropsychological testing, revealed a seizure onset in the nondominant temporal lobe. All patients had an excellent outcome after epilepsy surgery. Peri-ictal water drinking may represent a lateralizing sign indicating seizure onset in the nondominant temporal lobe. PMID- 12629254 TI - Paraplegia after ligation of esophageal varices. PMID- 12629253 TI - Thalidomide-induced neuropathy: a ganglionopathy? PMID- 12629255 TI - Lyme disease and optic neuritis: long-term follow-up of seropositive patients. PMID- 12629256 TI - Zolmitriptan for treatment of migrainous vertigo: a pilot randomized placebo controlled trial. PMID- 12629257 TI - Minocycline for Huntington's disease: an open label study. PMID- 12629258 TI - Effects of subthalamic nucleus (STN) stimulation on motor cortex excitability. PMID- 12629259 TI - Unusual entrapment neuropathy in a golf player. PMID- 12629260 TI - Predictors of effective bilateral subthalamic nucleus stimulation for PD. PMID- 12629261 TI - Plasma susceptibility to free radical-induced antioxidant consumption and lipid peroxidation is increased in very old subjects with Alzheimer disease. AB - Oxidative stress is believed to play a major role in the pathogenesis of Alzheimer disease (AD). Plasma concentrations of vitamins C, A and E, of uric acid, thiols and carotenoids were lower and of malondialdehyde (MDA) higher in 35 AD patients (85.9 +/- 5.5 y) compared to 40 controls 85.5 +/- 4.4 y). Differences were significant for vitamin C, vitamin E, lutein, lycopene, alpha-carotene and MDA (p < 0.001). Plasma exposed to peroxyl radicals showed a rate of antioxidant consumption and of MDA production higher in AD patients than in controls. AD in advanced age is accompanied by a poor plasma antioxidant status and increased plasma lipid peroxidation, as well as by a low resistance to peroxyl radical exposure. PMID- 12629263 TI - The quandary of reductionism: relevance to Alzheimer disease research. AB - Modern science has embraced reductionism, seeking ever-smaller parts to explain the whole. Although reductionistic approaches are successful in very simple biological modelling, they are not necessarily appropriate for systems of increasing complexity. Drawing on famous historical examples of how non reductionist thinking has benefited mankind, and of how reductionism has sometimes led to erroneous conclusions, we call attention to the need to move away from purely linear reasoning in order to succeed in addressing many of the problems we face with the predicted demographic increase in seniors, and the increase in numbers of those afflicted with Alzheimer disease. The time has come to reconsider and seriously question our most basic assumptions and beliefs surrounding what we believe Alzheimer disease to be, without which we run the risk of missed opportunities and failure. PMID- 12629264 TI - From acute to chronic pancreatitis: the role of mutations in the pancreatic secretory trypsin inhibitor gene. AB - Pancreatic secretory trypsin inhibitor (PSTI) is a potent natural inhibitor of trypsin. We proposed the hypothesis that, if the function of the PSTI is impaired by its genetic mutation, trypsin may easily promote autodigestion causing pancreatitis and we performed a mutational analysis of the PSTI gene in patients with pancreatitis. Two exonic mutations (N34S and R67C) were thought to be associated with a predisposition to pancreatitis. The N34S mutation was co segregated with two intronic mutations, IVS1-37T>C and IVS3-69insTTTT. Although we analyzed the function of the recombinant N34S protein, we could not demonstrate the loss of function of this protein. Intronic mutations, rather than N34S itself (IVS1-37T>C + N34S + IVS3-69insTTTT complex), may be associated with the decreased function of the PSTI. Alternatively, increased digestion of N34S in vivo may be applicable. As for R67C, the conformational alteration of the protein by forming intra-molecular or inter-molecular disulfide bonds with 67Cys was strongly suggested. These results, along with the brand-new findings in PSTI knockout mice, suggest that the genetic mutation of the PSTI is one of the important mechanisms for predisposition to pancreatitis by lowering the trypsin inhibitory function. PMID- 12629265 TI - Pancreatic renin-angiotensin system: a novel target for the potential treatment of pancreatic diseases? PMID- 12629266 TI - An insight into the role of human pancreatic lithostathine. AB - Human lithostathine was initially isolated from pancreatic stones in patients with alcoholic calcifying chronic pancreatitis. It is secreted into the pancreatic juice where it was believed to inhibit stone formation. The N-terminal undecapeptide was assumed to play an important role in the mechanism, by adsorption to the crystal surface. Later, the role of lithostathine in calcite formation and growth was questioned, together with the associated mechanism of action. In particular, although lithostathine adsorbs on calcite crystal, this property does not now seem to be specific. Moreover, the N-terminal undecapeptide is not likely to have, by itself, the function of the entire protein. The different aspects of this controversy are reviewed and discussed, particularly in the light of recent structural biology. Comparative biological data now available allow us to draw an interesting parallel between lithostathine and other related proteins. Finally, lithostathine might affect stone formation and may also have another function which could be investigated in the other proteins belonging to the same structural family. PMID- 12629267 TI - Acute pancreatitis secondary to pancreatic neuroendocrine tumours. AB - CONTEXT: Pancreatic neoplasms are an uncommon aetiology of acute pancreatitis. Pancreatic neuroendocrine tumours are a rare subgroup of pancreatic neoplasms. CASE REPORT: We report on three patients having acute pancreatitis secondary to pancreatic neuroendocrine tumours, one of them with severe pancreatitis, and review the published cases up to now. Only 22 patients with acute pancreatitis secondary to pancreatic neuroendocrine tumours have been reported (including the present cases). Most of these cases were of non-functioning neoplasms and the course of the pancreatitis tended to be mild. In the most recent reports and in the present cases, the initial diagnostic method was CT scan. Less than half had metastases when the tumour was diagnosed and mortality from these neoplasms reached approximately 50%. CONCLUSIONS: Pancreatic neuroendocrine tumours can cause acute pancreatitis even in patients under 50 years of age. On many occasions, the tumours are non-functioning; therefore, acute pancreatitis may be the first clinical symptom. Consequently, faced with acute pancreatitis of unknown origin, a non-functioning neuroendocrine tumour should be ruled out. PMID- 12629268 TI - The curious genomic path from leaky red cell to nephrotic kidney. AB - The human red cell has proved to be an invaluable model cell for the study of many aspects of membrane structure and function. It has a series of transport pathways which mediate the movements of the univalent cations Na and K, which are either identical or similar to systems in other human tissues, including the human kidney. The balance between the energy-consuming NaK pump and a 'passive leak' component maintains a net deficit of cations within the cell, which defends the cell volume against osmotic swelling. There exist a series of dominantly inherited human red cell conditions, gathered under the generic title 'hereditary stomatocytoses', in which the so-called 'passive leak' to Na and K is pathologically increased. In the more severe variants this compromises the integrity of the cell and the patients suffer haemolytic anaemia. Some less severe variants present with pseudohyperkalaemia caused by loss of K from red cells on storage of blood at room temperature. The most severe variants show a deficiency in a widely distributed 'raft' protein known as stomatin. The stomatin protein is homologous to the 'podocin' protein, the gene for which is mutated in a recessively inherited form of nephrotic syndrome. Among other possible functions, both proteins could be involved in the trafficking of membrane proteins to and from the plasma membrane. PMID- 12629269 TI - Urinary concentration defect and limited expression of sodium cotransporter, rBSC1, in a rat model of chronic renal failure. AB - BACKGROUND: Renal urinary concentration is associated with enhanced expression of sodium cotransporter (rBSC1) in thick ascending limb of Henle. Overexpression of rBSC1 was reported recently in hypertrophied nephrons after unilateral nephrectomy (UNX) and in kidney isografts. Since urinary concentration defect and hypertrophy of residual nephrons are major manifestations of chronic renal failure (CRF), we investigated the rBSC1 signals for RNA and protein in a rat model of CRF. METHODS: Rats underwent 5/6 nephrectomy and examined 8 weeks after operation. rBSC1 mRNA was examined by competitive PCR and in situ hybridization, and rBSC1 protein signals by western blotting and immunohistochemistry. Rats that underwent sham-operation, UNX, or 5/6 nephrectomy followed by a 3-week recovery period (acute renal failure), were used as control. Water intake was restricted for 24 h in subgroups of control and CRF rats. RESULTS: Microscopic examination showed hypertrophy of residual nephrons in both UNX and CRF rats. Signals for rBSC1 mRNA and protein were enhanced at basal condition only in rats with UNX. Under basal conditions, CRF rats demonstrated low urinary osmolality in spite of high plasma arginine vasopressin levels. Water restriction resulted in increased signals for rBSC1 mRNA and protein and concentration of urine in sham-operated rats, but such increases were absent and urinary concentration was incomplete in CRF rats. CONCLUSIONS: Compensatory overexpression and upregulation of rBSC1 expression in response to dehydration are both absent in CRF rats. These limitations are thought to be the underlying mechanisms of urinary concentrating defect seen in CRF. PMID- 12629270 TI - A nitric oxide-generating beta-blocking agent prevents renal injury in the rat remnant kidney model. Comparative study of two beta-blocking drugs, nipradilol and propranolol. AB - BACKGROUND: The L-arginine-nitric oxide (NO) pathway plays an important role in the modulation of glomerular disease. We investigated whether beta-blocking agents, with and without an NO-generating function, had renoprotective effects in the 5/6 nephrectomized rats (Nx), an animal model of glomerulosclerosis. METHODS: Nipradilol, a beta-blocker with an ONO(2) group (5, 10 or 15 mg/kg/day) and propranolol, a beta-blocker without this group (50 mg/kg/day) were administered for 12 weeks to Nx together with and without nitro-L-arginine methyl ester (L NAME). We evaluated the effects of both drugs on proteinuria, hypertension, renal function, glomerulosclerosis and urinary excretion of NO metabolites (U(NOx)) and cyclic GMP (U(cGMP)). RESULTS: Both drugs similarly attenuated the elevated blood pressure in Nx. However, nipradilol, at doses of 10 and 15 mg/kg/day, significantly decreased proteinuria and glomerulosclerosis, while propranolol did not. Nx showed reduced U(NOx) in comparison with the sham-operated rats. Nipradilol increased U(NOx) and U(cGMP) significantly and in a dose- dependent manner, whereas propranolol reduced them to levels lower than those in Nx. Nx receiving L-NAME reduced U(NOx). The addition of nipradilol increased U(NOx) and decreased urinary protein excretion and glomerulosclerosis, suggesting that the NO released from the drug contributed to its renoprotective effect. CONCLUSION: These findings indicate that nipradilol exerts its renoprotective effect through NO generation, and not by lowering blood pressure. The beta-adrenergic blocking action per se does not seem to be related to the renoprotective effect of these agents. PMID- 12629272 TI - Does bone marrow contain renal precursor cells? AB - In the past few years stem cells normally present within the bone marrow have been shown to have considerable plasticity in the repertoire of the cells that they produce. They are capable of producing specific lineages of cells not only of the blood, but also contribute usefully to lineages of other organs. In the kidney, studies in humans and rodents provide evidence for the presence of bone marrow-derived 're-programmed' cells, but it remains possible that the cells that arrive and re-differentiate are no longer stem cells (quiescent cells, not actively proliferating). Nevertheless, the fact that appropriately differentiated cells are delivered deep within the kidney simply by injection of bone marrow cells should make us think differently about the way these organs regenerate and repair. Migratory pathways for multipotential stem cells could be exploited to effect repair using an individual's own stem cells, perhaps after gene therapy. It is clear also that a transplanted organ would in time become affected by the genetic susceptibilities of the recipient, because of phenotypes that are expressed when trafficking cells engraft and differentiate. PMID- 12629271 TI - Reversible angiotensin II-mediated albuminuria in rat kidneys is dynamically associated with cytoskeletal organization. AB - Angiotensin II (ANG II) is intimately involved in normal renal function, and is estimated to exist at a normal physiological range of 6-10 nM within the renal tubules. The potential role that intrarenal ANG II may play in renal disease was assessed by perfusing isolated rat kidneys with or without excess intratubular levels of ANG II, which may mimic changes in the intrarenal RAS under pathological conditions. The effects of increased systemic ANG II were also determined by infusing rats with ANG II by osmotic pump. In isolated perfused kidneys, ANG II significantly and specifically increased the fractional clearance of albumin to clinical levels, as determined by using radiolabelled albumin. This effect was reversible, as removing ANG II from the perfusate caused the albumin fractional clearance to decrease to pre-ANG II exposure levels. The increase in fractional clearance of albumin was not correlated with renal hemodynamic changes, nor glomerular permeability alterations as measured by the fractional clearance of 36 A Ficoll and immunoglobulin G. Immunochemical analysis using anti alpha-tubulin antibody of perfused kidney sections revealed that ANG II caused a marked disruption of tubular epithelial cytoskeletal components, through disassembly and reorganization of alpha-tubulin. This disruption was reversible. In vivo, osmotic pump delivery of ANG II at less potent dosage caused a proteinuria (Biuret) and an albuminuria (radioimmunoassay) in rats, from as early as 2 days after pump implantation. These results demonstrate that ANG II may reversibly induce clinical levels of albuminuria. These data point to an important role for renal tubules and the intratubular lumen concentrations of ANG II in the renal processing of albumin. PMID- 12629273 TI - Therapeutic promise of embryonic kidney transplantation. AB - One novel solution to the shortage of human kidneys available for transplantation envisions 'growing' new kidneys in situ via xenotransplantation of renal anlagen. We and others have shown that developing metanephroi transplanted into animal hosts undergo differentiation and growth, become vascularized by blood vessels of host origin and exhibit excretory function. Metanephroi can be stored for up to 3 days in vitro prior to transplantation with no impairment in growth or function post-implantation. Metanephroi can be transplanted across both concordant (rat to mouse) and discordant/highly disparate (pig to rodent) xenogeneic barriers. Here we review studies exploring the potential therapeutic use of embryonic kidney transplantation. PMID- 12629274 TI - Phenotyping renal leukocyte subsets by four-color flow cytometry: characterization of chemokine receptor expression. AB - To investigate mechanisms of cell-mediated injury in renal inflammatory disease it is critical to determine the surface phenotype of infiltrating renal leukocyte subsets. However, the cell-specific expression of many leukocyte receptors is difficult to characterize in vivo. Here, we report a protocol based on flow cytometry that allows simultaneous characterization of surface receptor expression on different subsets of infiltrating renal leukocytes. The described technique combines an adapted method to prepare single cell suspensions from whole kidneys with subsequent four-color flow cytometry. We recently applied this technique to determine the differential expression of murine chemokine receptors CCR2 and CCR5 on infiltrating renal leukocyte subsets. In this article, we summarize our current findings on the validity of the method as compared with immunohistology and in situ hybridization in two murine models of nonimmune (obstructive nephropathy) and immune-mediated (lupus nephritis) inflammatory renal disease. Flow cytometry analysis revealed an accumulation of CCR5-, but not CCR2-positive lymphocytes in inflamed kidneys, compared to the peripheral blood. Particularly renal CD8+ cells expressed CCR5 (79% in obstructed kidneys, 90% in lupus nephritis). In both models, infiltrating renal macrophages were positive for CCR2 and CCR5. These data corresponded to immunohistological and in situ hybridization results. They demonstrate that flow cytometric analysis of single cell suspensions prepared from inflamed kidneys is a rapid and reliable technique to characterize and quantify surface receptor expression on infiltrating renal leukocyte subsets. PMID- 12629275 TI - Selective cyclooxygenase-2 inhibition upregulates renal cortical alpha V integrin expression. AB - BACKGROUND: Cyclooxygenase-2 (COX-2), the inducible isoform of the cyclooxygenases, is upregulated in various inflammatory renal diseases and responsible for prostaglandin formation. As prostaglandins are known to influence cell adhesion processes, we investigated the effect of COX-2 inhibition on the expression of alpha(v) integrins, which are also enhanced in renal diseases and control the adherence between the endothelium and the extracellular matrix (ECM) in the glomerulus. METHODS: Healthy female Wistar rats and animals with previously induced passive Heymann nephritis (PHN) received either 5 mg/kg body weight/day celecoxib or a placebo. After 28 days, renal cortical mRNA expression of COX-2 and alpha(v) integrin subunits was determined. RESULTS: Rats with PHN showed a significant 1.7-fold increase in renal cortical mRNA expression of alpha(v) integrin subunits. Treatment with celecoxib increased cortical alpha(v) integrin mRNA expression 2.2-fold (p < 0.05) in healthy animals and 4.0-fold (p < 0.05) in rats with PHN, but lowered COX-2 mRNA expression in rats with PHN to 0.8 fold (p < 0.05). An inverse correlation between the expression of COX-2 and alpha(v) integrins in rats with PHN was demonstrated. CONCLUSIONS: It is suggested that COX-2-derived prostaglandins suppress the expression of alpha(v) integrins. This implies a previously unknown role for COX-2 in chronic inflammation in the kidney. PMID- 12629276 TI - Expression and cellular localisation of renal endothelin-1 and endothelin receptor subtypes in autosomal-dominant polycystic kidney disease. AB - The major factors influencing the rate of progression of chronic renal disease in autosomal-dominant polycystic kidney disease (ADPKD) are unknown and there are currently no effective treatments for slowing the progression of chronic renal failure in ADPKD patients. As a first step in investigating the potential role of endothelin-1 (ET1) and its receptors (ETA and ETB) in the pathophysiology of progression in ADPKD, we have studied their expression and cellular localisation in ADPKD kidney. Immunoreactive ET1 was detected in cyst epithelia, mesangial cells and vascular smooth muscle cells suggesting continuing ET1 synthesis in the cystic kidney. Compared to healthy controls, ETA mRNA was 5-10-fold higher in ADPKD cystic kidney. In cystic kidney, neo-expression of ETA receptors was found overlying glomeruli and cysts and markedly increased in medium-sized renal arteries by microautoradiography. This is the first study to demonstrate a specific upregulation of ETA receptors in human renal disease. Future studies should address whether ETA selective antagonists may be effective in slowing renal disease progression in ADPKD. PMID- 12629277 TI - Atrial fibrillation is associated with severe acute ischemic stroke. AB - Atrial fibrillation (AF) is a common risk factor for disabling ischemic stroke in the elderly, but it is not clear that its severity is generally worse than that of ischemic stroke due to other etiologies. We reviewed the clinical presentations of patients with acute ischemic stroke admitted between 1990 and 2001. The etiologies of these strokes were also classified using well-established criteria. Of 1,061 patients with acute ischemic stroke, 216 (20.3%) had AF. The frequency of bedridden state was 41.2% in patients with AF, compared to 23.7% in patients without AF (p < 0.0005). Other measures of clinical stroke severity showed similar disparities between these groups. The odds ratio for bedridden state following ischemic stroke due to AF was 2.23 (95% CI = 1.87-2.59, p < 0.0005) by multivariate logistic regression. Ischemic stroke associated with AF is typically more severe than ischemic stroke due to other etiologies, and this increased severity is independent of advanced age and other stroke risk factors. PMID- 12629278 TI - Vestibular schwannoma, tinnitus and cellular telephones. AB - Cases with tinnitus after using analogue cellular telephones are presented. An increased odds ratio of 3.45, 95% confidence interval (CI) 1.77-6.76, was found for vestibular schwannoma (VS) associated with the use of analogue cell phones. During the time period 1960-1998, the age-standardized incidence of VS in Sweden significantly increased yearly by +2.53% (CI 1.71-3.35). A significant increase in the incidence of VS was only found for the latter of the two time periods 1960 1979 and 1980-1998. For all other brain tumors taken together, the incidence significantly increased yearly by +0.80% (CI 0.59-1.02) for the time period 1960 1998, although the increase was only significant for benign tumors other than VS during 1960-1979. PMID- 12629279 TI - Laterality of brain tumors. AB - Tumor laterality was evaluated with respect to presenting symptoms and demographic factors among 489 adults with histologically confirmed glioma (354 high-grade, 135 low-grade), 197 with meningioma, and 96 with acoustic neuroma. The ratio of left-sided to right-sided tumors did not differ significantly from 1.00 for any of the major tumor types. Low-grade glioma and meningioma occurred nonsignificantly more often on the left side, whereas high-grade glioma and acoustic neuroma occurred nonsignificantly more often on the right side. Aphasia or mental status changes were significantly more common among glioma patients with tumors on the left side than among those with tumors on the right side. Associations between tumor laterality and symptoms may influence the probability or timing of diagnosis, possibly differentially by marital status. PMID- 12629280 TI - High prevalence of calcified silent neurocysticercosis in a rural village of Mexico. AB - Human neurocysticercosis (NC) is a parasitic disease caused by TAENIA SOLIUM when its larvae lodge in the central nervous system. NC prevalence estimates are obscured by the variable and often asymptomatic clinical picture. While infection depends on exposure, severity is possibly related with various host factors (immunity, genes and gender). This epidemiological study of cranial CT scans in an endemic rural community found that 9.1% of apparently healthy subjects had calcified lesions and were completely asymptomatic. Silent NC cases did not correlate with the exposure factors tested but showed family aggregation and higher rates of positive serology. Thus, NC prevalence may be higher than currently considered and host-related factors appear to be involved in infection and pathogenesis. PMID- 12629282 TI - Is carotid endarterectomy effective and safe in asymptomatic patients with carotid stenosis? PMID- 12629281 TI - Elevated suicide risk among patients with multiple sclerosis in Sweden. AB - Results from previous studies of suicide risk among patients with multiple sclerosis (MS) are inconsistent. This may be explained partly by differences in methodology and study populations. The purpose of our study was to investigate suicide risk among hospital patients with MS in Sweden. During the period 1969 1996, 12,834 cases were recorded in the Swedish Hospital Inpatient Register, with 77,377 hospital admissions, in which MS was a primary or secondary diagnosis at discharge. The mean follow-up time for the whole cohort was 9.9 (SD 7.3) years. When the data for these MS patients were linked to the Swedish Causes of Death Register for the same period, 5,052 (39.4%) were found to have died. Among the 5,052 deaths, suicide was an underlying cause of death in 90 cases (1.8%). The mean period between the initial admission date with an MS diagnosis at discharge and the date of death for the 90 MS suicide cases was 5.8 (SD 5.1) years. This was significantly shorter (p = 0.002) than the mean of 7.9 (SD 6.4) years for MS cases who died due to other causes. Suicide risk, calculated as the standardized mortality ratio (SMR), was significantly elevated (SMR = 2.3) among both male and female MS cases compared with the general population. Suicide risk was particularly high in the first year after initial admission with an MS diagnosis, and among younger male MS cases. The mean age at the time of suicide was 44.5 (SD 12.4) years, and 58% of the suicides were committed within 5 years after the first admission with an MS diagnosis. The crude suicide rate among MS patients during the study period was 71 per 100,000 person-years. The rate was significantly higher (p < 0.001) in males (114) than in females (47), with an odds ratio of 2.4 (95% CI: 1.6-3.8). These findings have implications for suicide preventive measures in neurological practice. PMID- 12629284 TI - Screening for alcohol use. PMID- 12629288 TI - Going with the flow of anticoagulant therapy. PMID- 12629289 TI - Keeping up with standards: your key to safe practice. PMID- 12629291 TI - Electroconvulsive therapy sheds its shocking image. PMID- 12629292 TI - Burn care basics: How to extinguish problems. PMID- 12629293 TI - Patient-education guide. Depression. PMID- 12629294 TI - Name that tube. PMID- 12629296 TI - Benzocaine-induced methemoglobinemia. PMID- 12629297 TI - Partners in action: nursing education and nursing practice. PMID- 12629299 TI - Grievances and complaints: valuable tools for management and for staff. PMID- 12629298 TI - Raising the bar and keeping it there. PMID- 12629300 TI - Implementing advanced nurse practitioner roles in acute care: an evaluation of organizational change. AB - Medical shortages are seen as the driving force behind the recent implementation of the advanced nurse practitioner (ANP) role in some Canadian hospitals. The authors analyzed the implementation of the ANP role in one tertiary care teaching hospital from the organizational change perspective. Despite successful implementation, issues of role definition, scope of practice boundaries, and staff reactions remain unresolved. Recommendations are discussed for implementing ANP roles in other acute care facilities. PMID- 12629302 TI - Workplace prevention and musculoskeletal injuries in nurses. AB - OBJECTIVE: The purpose of this study was to describe the availability of preventive devices and training in relation to neck, shoulder, and back musculoskeletal injuries/disorders (MSD) in registered nurses. SUMMARY BACKGROUND DATA: Nurses have one of the highest rates of MSD of any occupation. Studies have shown that mechanical lifting devices and lifting teams can reduce MSD rates and associated costs. METHODS: Data from 1163 randomly selected currently working nurses (1+ years on the current job) were collected in anonymous mailed surveys (74% response rate). MSD cases had neck, shoulder, and/or back symptoms for at least 1 week, or at least monthly, and moderate or more pain, in the past year. RESULTS: Nurses with mechanical lifting devices available were significantly less likely to have neck or back MSDs. Back injury was less likely when lifting teams were available. However, adjustable beds and transfer sheets were associated with greater odds of back MSD. Training in workstation adjustment was associated with significantly lower MSD prevalence, though postural training was not. CONCLUSIONS: Though use of mechanical devices and lifting teams was limited in nursing workplaces, these prevention strategies were related to reduced odds of MSD. Nursing administrators can use these findings to consider workplace changes. PMID- 12629301 TI - A clinical ladder program based on Carper's Fundamental Patterns of Knowing in Nursing. AB - Clinical and career ladder programs were popular adjuncts to recruitment and retention during the nursing shortage of the 1980s. Programs commonly used Benner's work as an organizing framework and typically used activities such as continuing education credit, committee participation, work experience, certifications, academic degrees, community service, and performance appraisal scores as criteria for advancement. The authors present a unique clinical ladder program based on Carper's Fundamental Patterns of Knowing in Nursing. This philosophical work has been debated in the literature for many years, yet to the authors' knowledge, this work has never been used as the conceptual basis for program development. The four patterns of knowing in nursing were found to be consistent with the definition of professional nursing, and provided a meaningful organizing framework that was well received by staff registered nurses. An overall history of clinical ladders is presented, followed by the organization's prior experience with these programs, and finally the details of development and implementation of the program with plans for evaluation. PMID- 12629304 TI - Job and career satisfaction among staff nurses: effects of job setting and environment. AB - Just as customer satisfaction is the key to retaining customers, satisfaction with job and career choices are important for keeping staff nurses on the job. The roles of employment setting, job commitment, tenure, years until retirement, short staffing, and patient load in predicting satisfaction were assessed for RN and LPN staff nurses. Results show that when RNs and LPNs feel short staffing interferes with their ability to meet patient care needs, they are also less satisfied with both their job and their career. In order not to exacerbate the current nursing shortage, employers must find ways to ensure adequate staffing to keep staff nurses satisfied and on the job. PMID- 12629303 TI - Measuring practice patterns among acute care nurse practitioners. AB - OBJECTIVE: This initiative was designed to develop a reliable instrument to measure the activities of acute care nurse practitioners (ACNPs). A sound, standardized method for measuring ACNP productivity will assist nursing leaders and administrators to demonstrate the effectiveness and productivity of ACNPs in and across institutions and systems. BACKGROUND DATA: Current research on ACNPs uses many different methodologies and research designs, and fails to provide standard definitions to measure practice patterns, making it difficult to generalize across settings. METHODS: Advisory groups from 2 New York academic health science centers developed a survey that covered the demographic, educational, and employment characteristics of ACNPs, and a 20-item classification of advanced practice nursing activities. Sixty-one ACNPs completed surveys, a 58% response rate. RESULTS: The survey found strong similarities at both institutions. ACNPs spend most of their time in 5 activities involving direct care and 4 activities within indirect care. Strong Cronbach alphas confirmed that the instrument was reliable. CONCLUSIONS/IMPLICATIONS: The availability of a reliable instrument for measuring ACNP practice patterns provides administrators with a powerful tool to demonstrate the contributions of their ACNPs. In addition, a standardized method for data collection can contribute to healthcare workforce policy discussions. PMID- 12629305 TI - A different age for leadership, part 2: new rules, new roles. AB - This is Part 2 of a 2-part article on the new and emerging characteristics and elements of leadership for changing, fast-paced organizations. As we leave the 20th century workplace and are increasingly driven by innovation and technological transformation, new roles are demanded from everyone. Leadership expression now calls for a different emphasis and skill-set from those that predominated in the past. The first article (February 2003) focused on the context of leadership affecting what leaders do and how they must now behave. This article centers on the activities of leadership and the new learning and skill-set development that will increasingly be required of leaders in a fundamentally altered work environment. PMID- 12629306 TI - The best of both worlds: a consideration of gender in team building. AB - As teams continue to supplant individuals as the fundamental work unit, healthcare organizations increasingly are turning to high-performance care and service groups as keys to success. In this article, the author presents research and findings of an ethnographic case study of a mixed-gender work team of healthcare administrators. Focusing on gender issues and their impact on outcomes, the study offers insights for promoting cultural change and improving nursing practice systems that will influence health policy and support a new architecture for the healthcare industry. PMID- 12629307 TI - A larger worldview. PMID- 12629308 TI - Is fetal pulse oximetry ready for clinical practice?: Writing for the PRO position. PMID- 12629309 TI - Is fetal pulse oximetry ready for clinical practice?: Writing for the CON position. PMID- 12629310 TI - Personal data assistants: using new technology to enhance nursing practice. AB - This article explains how the new technology of personal data assistants can be used to enhance and augment comprehensive nursing care. Nurses are constantly challenged in their need for current, reliable, and accurate information at the point of patient care. Professional books and journals, by the very nature of their print format, have been prepared long before they can be actually used in practice. More current information is available from the World Wide Web, but it is often impractical for a nurse to access a computer during a patient encounter. Personal data assistants [PDAs] allow clinicians to access and document absolutely current information at the moment the patient is being seen. There are many general applications for PDAs that nurses might use such as keeping electronic calendars, address books, and reminder lists. In addition, however, there are even more actual healthcare applications, including patient tracking systems, access to pharmacologic databases, and a variety of clinical decision making support tools. This article describes the wide variety of PDAs, along with the factors a nurse should consider in the decision of whether to purchase a PDA, and which type of device is best suited for which application. PMID- 12629311 TI - Postpartum beliefs and practices among non-Western cultures. AB - Postpartum health beliefs and practices among non-Western cultures are each distinct, but have many similarities. Two common belief systems surround 1) the importance of hot and cold, and 2) the necessity of confinement during a specific period of time after giving birth. This article describes common postpartum health beliefs among women in Guatemala, China, Jordan, Lebanon, Egypt, Palestine, India, and Mexico, and offers an exemplar from the author's experiences as a Korean woman giving birth in the United States. Cultural competence in the provision of postpartum care is essential for nurses in the healthcare world of the 21st century. PMID- 12629312 TI - Self-care in Chinese school-age children with nephrotic syndrome. AB - PURPOSE: To identify the levels of self-care in Chinese school-age children with primary nephrotic syndrome. STUDY DESIGN METHODS: Descriptive study of 66 school age children hospitalized at least once for nephrotic syndrome who were followed medically at the affiliated hospitals of Sun Yat-sen University. Demographic data and information about nephrotic syndrome self-care were obtained by interviewing patients' families. Frequency distributions were used for the analysis. RESULTS: Of 66 patients studied, 60 had high levels of self-care practices. High levels of self-care were found for 83% of 6 to 8 year olds, 95% of 9 to 10 year olds, all 11 to 12 year olds. CLINICAL IMPLICATIONS: This study can help nurses to understand cross-cultural implications of self-care practices for Chinese school age children. The findings also can guide nurses to assess promote self-care practices of pediatric patients. PMID- 12629313 TI - School violence: an insider view. AB - PURPOSE: To discover what teachers perceive to be contributing factors to violence in schools. STUDY DESIGN AND METHODS: Open-ended questions were asked of a convenience sample of teachers ( = 396) during an in-service education program on school violence. The teachers were in a semi-rural school district in a Mid Atlantic state. Answers were analyzed using content analysis; all responses were reviewed and important themes were extracted. Identified themes were then placed into suitable categories and studied to determine relationships. RESULTS: Of the surveys analyzed ( = 239), 13 themes were identified. The three categories which then identified probable causes of school violence were (1) lack of knowledge, (2) lack of support, and (3) inadequate safety measures. CLINICAL IMPLICATIONS: Nurses can use the results of this study in multiple ways. One is to help parents understand their role in preventing school violence. Because violence in the home and violence in the media seem to foster violent acting-out behavior, nurses can teach parents about these correlations and seek solutions such as the elimination of family violence, and monitoring television viewing and video games. Nursing assessments of school-aged children and their families can include these elements. School nurses in particular can use these study results as an opportunity to develop interventions for students, teachers, and families that stress knowledge building about impulse control, anger management, appropriate parenting, and early intervention for at-risk children. PMID- 12629314 TI - Adolescent motherhood: a meta-synthesis of qualitative studies. AB - PURPOSE: To synthesize qualitative studies on the phenomenon of adolescent motherhood. METHODS: Databases were searched for qualitative studies regarding the experience of adolescent motherhood (years 1990-2001). Noblit and Hare's (1988) meta-ethnographic comparative method was used for synthesizing the research. A total of 251 citations were found, with a subgroup of 50 studies on "adolescent motherhood" and "qualitative studies." Twenty-five studies conducted in the United States, Canada, China, England, and Australia met the sample inclusion criteria. The combined sample of participants across studies was 257 adolescent mothers. RESULTS: Certain characteristics are common to the studies of adolescent motherhood. Five overarching metaphors were found: (1) the reality of motherhood brings hardship; (2) living in the two worlds of adolescence and motherhood; (3) motherhood as positively transforming; (4) baby as stabilizing influence; and (5) supportive context as turning point for the future. CLINICAL IMPLICATIONS: Nurses working in hospital, home care, community, and school settings can use the results of this metasynthesis to obtain a clearer picture of how adolescents feel during pregnancy, and the types of nursing interventions that can be developed to help them. PMID- 12629315 TI - Caring for the infertile woman. AB - Increasing numbers of women are seeking infertility services. In light of this, it is important that professional nurses learn about infertility treatments and options, for the public looks to nurses for guidance in all aspects of healthcare. This article describes the systematic assessment, planning, implementation, and evaluation phases of the nursing process for women who are involved in infertility evaluation and interventions. Much of the basic work-up for infertility can be accomplished by a generalist (OB/GYN, CNM, or nurse practitioner), but if the patient does not conceive in a few months, referral to an infertility specialist is essential. Women need help to understand the possibilities as well as the limitations of current therapies for infertility. The toll that the emotional roller coaster of infertility takes on women and their partners can be devastating so those caring for them must be prepared to address that aspect of the experience. The nurse must be prepared to help the infertile woman deal with crushing disappointment and grief when cycles fail or pregnancies end in the early stages. The caring aspect of professional nursing is an essential component of meeting the special needs of these patients. PMID- 12629316 TI - The relationship between abuse, sexually transmitted diseases, & group B streptococcus in childbearing women. AB - PURPOSE: To examine the relationship between abuse, sexually transmitted diseases (STDs) and group B (GBS) infection among childbearing women using Selye's (1978) stress response theory. DESIGN AND METHODS: Retrospective chart review (n = 205) from two different clinical sites in Washington State, using the Childbearing Health Questionnaire to guide data collection. The women in the sample had an average age of 26.4 years and represented Anglo (81.4%), Hispanic (12%), Native American (3.9%), and African American (2.5%) ethnic groups. Thirty-eight percent ( = 78) reported experiencing physical and/or sexual abuse during their lifetimes and 31% had been diagnosed with an STD. RESULTS: Abuse was significantly related to STDs, and ethnicity emerged as a significant variable for the Hispanic women participating in this study. Findings indicated that infection with group B was also related to abuse status ( r=.60, p < or =.002) and to presence of herpes simplex virus-2 (r =.468, p0.8). CONCLUSIONS: In a comprehensive analysis of variables reported to influence lower pole stone clearance artificial neural network analysis predicted stone clearance with a high degree of accuracy. The relative importance of dynamic urinary transport in lower pole stones and the usefulness of artificial neural network analysis to predict shock wave lithotripsy outcomes in individuals must be confirmed in a prospective trial. PMID- 12629338 TI - A prospective randomized controlled trial on ureteral stenting after ureteroscopic holmium laser lithotripsy. AB - PURPOSE: A prospective randomized controlled trial was conducted to evaluate whether postoperative ureteral stenting is necessary after ureteroscopic laser lithotripsy. MATERIALS AND METHODS: A total of 58 patients with unilateral ureteral stones were randomized into either stented or unstented groups. Ureteroscopic laser lithotripsy was performed using a semirigid ureteroscope (6.5/7Fr) and holmium laser without ureteral orifice dilation. There were no selection criteria regarding stone size, location, preoperative ureteral obstruction and hydronephrosis. Endoscopic evidence of stone impaction or mucosal edema/damage did not exclude a patient from the study. Ureteral perforation on completion retrograde pyelogram was the only intraoperative criterion for study exclusion. Postoperative pain scores and symptoms were recorded. Excretory urography was performed to document stone-free status and stricture formation. Radionuclide scan was performed selectively to exclude functional obstruction when ureteral narrowing was found on excretory urogram. RESULTS: Mean stone size +/- SD was 9.7 +/- 4.0 mm. (range 4 to 27). Proximal ureteral stones accounted for 43% of all stones. Stented and unstented groups were comparable with respect to demographic data, stone parameters, preoperative obstruction and hydronephrosis. There was no significant difference in operating time, laser energy used, stone impaction and mucosal edema/damage between the 2 groups. Postoperative pain and symptoms were more severe and frequent (p <0.05) in the stented group. However, there was no difference in the incidence of postoperative sepsis and unplanned medical visits. The stone-free and stricture formation rates showed no statistical difference between the 2 groups. CONCLUSIONS: Ureteral stenting is not necessary after uncomplicated ureteroscopic laser lithotripsy for ureteral stones. Ureteral stent increases the incidence of pain and urinary symptoms but does not prevent postoperative urinary sepsis and unplanned medical visits. Severity of preoperative obstruction and intraoperative ureteral trauma were not shown to be determining factors for stenting. PMID- 12629339 TI - Laparoscopic radical prostatectomy: oncological evaluation after 1,000 cases a Montsouris Institute. AB - PURPOSE: We performed a prospective oncological evaluation of laparoscopic radical prostatectomy in regard to local tumor control and biochemical recurrence. MATERIALS AND METHODS: Between January 1998 and March 2002, 1,000 consecutive patients with a mean age +/- SD of 63 +/- 6.2 years and clinically localized prostate cancer underwent laparoscopic radical prostatectomy at 1 institution. Preoperative 1997 TNM clinical stage was T1a in 6 patients (0.6%), T1b in 3 (0.3%), T1c in 660 (66.5%), T2a in 304 (30.4%) and T2b in 27 (2.7%). Mean preoperative prostate specific antigen (PSA) +/- SD was 10 +/- 6.1 ng./ml. (range 1.5 to 55). Postoperatively, surgical specimens were assessed and positive surgical margins recorded. Factors that could influence the surgical margins status were evaluated. Irrespective of pathological stage or surgical margin status, no adjuvant treatment was proposed before an increasing PSA. PSA recurrence was defined as PSA greater than 0.1 ng./ml. and was confirmed by a second increase. Recurrence time was defined as the time of the first increase in PSA. RESULTS: Postoperative pathological stage was pT2aN0/Nx in 203 patients (20.3%), pT2bN0/Nx in 572 (57.2%), pT3aN0/Nx in 142 (14.2%), pT3bN0/Nx in 77 (7.7%) and pT1-3 N1 in 6 (0.6%). Positive surgical margin rate was 6.9%, 18.6%, 30% and 34% for pathological stages pT2a, pT2b, pT3a and pT3b, respectively (p <0.001). The main predictors of a positive surgical margin were preoperative PSA (p <0.001), clinical stage (p = 0.001), pathological stage (p <0.001) and Gleason score (p = 0.003). The overall actuarial biochemical progression-free survival rate was 90.5% at 3 years. According to the pathological stage, the progression free survival rate was 91.8% for pT2aN0/Nx, 88% for pT2bN0/Nx, 77% for pT3aN0/Nx, 44% for pT3bN0/Nx and 50% for pT1-3N1 (p <0.001). Of the patients 94% with negative surgical margins and 80% with positive margins had progression-free survival (p <0.001). Preservation of the neurovascular bundles in patients with localized tumors had no significant effect on the subsequent risk of positive surgical margins or progression-free survival. CONCLUSIONS: Based on followup, our evaluation confirms that laparoscopic radical prostatectomy provides satisfactory results in regard to local tumor control and biochemical recurrence. PMID- 12629340 TI - Dynamic assessment of angiogenesis in renal cell carcinoma spheroids by intravital microscopy. AB - PURPOSE: Renal cell cancer represents a suitable tumor model for in vivo observation of neo-angiogenesis. We used intravital microscopy and the well established dorsal skin fold chamber model to characterize neo-angiogenesis in freely implanted renal cell cancer spheroids. MATERIAL AND METHODS: Tumor spheroids were implanted into dorsal skin fold chambers of 8 nude mice. At days 3, 6, 10 and 14 after implantation the newly vascularized spheroid area, density of perfused microvessels in the spheroid versus the periphery, capillary center erythrocyte velocity and capillary diameter were recorded by intravital microscopy. Video images were analyzed by a computer assisted image analysis device. After the experiments the chambers were analyzed morphologically. RESULTS: The model enabled quantitative analysis of microcirculation and angiogenesis in the renal cell cancer spheroids during 14 days of observation. Mean spheroid center perfused microvessel density +/- SEM increased from 3 +/- 2 to 269 +/- 21 cm.-1 on days 3 to 10 and subsequently decreased to 189 +/- 38 cm. 1 on day 14. Spheroid periphery perfused microvessel density was significantly higher throughout the experiments, attaining a mean maximum of 522 +/- 34 cm. on day 14. Mean capillary diameter decreased continuously from 14.2 +/- 0.9 to 8.4 +/- 0.4 microm. on days 3 to 14. In contrast, mean capillary center erythrocyte velocity significantly increased during 14 days of observation from 0.09 + 0.02 mm. per second on day 3 to 0.24 +/- 0.08 mm. per second on day 14. Histological analysis after 14 days revealed the spheroids as cell clusters in the upper layers of the dorsal skin fold chamber. CONCLUSIONS: The model is suitable for the analysis of renal cell cancer angiogenesis. Although it is heterotopic, angiogenesis in renal cell cancer spheroids mimics important characteristics of human renal cell cancer. PMID- 12629341 TI - Familial papillary renal cell tumors and subsequent cancers: a nationwide epidemiological study from Sweden. AB - PURPOSE: Familial risks in papillary renal cell carcinoma and association with second primary malignancies were studied using the nationwide Swedish Family Cancer Data Base. MATERIALS AND METHODS: Cancer data obtained from the Swedish Cancer Registry from 1961 to 1998 included 1,733 cases of papillary renal cell carcinoma. The standardized incidence ratio was used to measure cancer risk. RESULTS: Only 5 families were identified in which a parent had papillary renal cell carcinoma and an offspring had kidney cancer (nonsignificant SIR 1.51 for offspring). Discordant tumor sites associated with papillary renal cell carcinoma in the 2 generations were the upper aerodigestive tract and bladder (SIR 2.53, 95% CI 1.08 to 4.58 and 2.14, 95% CI 1.02 to 3.68, respectively). There was an overall increase in the risk of second primary malignancies of the lung, prostate and bladder and for non-Hodgkin's lymphoma and leukemia in patients with papillary renal cell carcinoma. The risk for a second primary tumor of the bladder associated with papillary renal cell carcinoma during the followup of 1 to 10 years was about 15 times higher than that associated with adenocarcinoma, which is the most common histological type of kidney cancer. The SIR was significantly higher in females than in males (59.67, 95% CI 40.23 to 82.94 versus 18.76, 95% CI 14.51 to 23.56). CONCLUSIONS: In addition to the familial association of these 2 cancer sites, the high risk of a second primary cancer of the bladder in patients with papillary renal cell carcinoma may reflect a common genetic alteration. PMID- 12629342 TI - The effect of bilaterality, pathological features and surgical outcome in nonhereditary renal cell carcinoma. AB - PURPOSE: We evaluated the differences in cancer specific, distant metastasis-free and local recurrence-free survival in patients with sporadic subtype concordant bilateral synchronous renal cell carcinoma and those with unilateral renal cell carcinoma, controlling for the covariates of subtype, stage, tumor size, grade and necrosis. We also analyzed early surgical complications and long-term renal function in patients who underwent staged surgery and those who underwent a single operation for bilateral synchronous renal cell carcinoma. MATERIALS AND METHODS: We retrospectively evaluated 44 patients with sporadic subtype concordant bilateral synchronous renal cell carcinoma treated at our institution between 1970 and 1998. There were 32 patients with bilateral synchronous clear cell renal cell carcinoma and 12 with bilateral synchronous papillary renal cell carcinoma. These patients were compared with 1,714 with sporadic unilateral clear cell renal cell carcinoma and 322 with sporadic unilateral papillary renal cell carcinoma treated with partial or radical nephrectomy during that period. Outcomes were estimated using the Kaplan-Meier method and Cox proportional hazard models were used to test associations with outcome. RESULTS: Clinicopathological features were similar for patients with bilateral synchronous and unilateral renal cell carcinoma except for the incidence of multifocality, which was 28% and 33% for bilateral synchronous clear cell and papillary renal cell carcinoma compared with 2% and 7% for unilateral clear cell and papillary renal cell carcinoma, respectively. Cancer specific survival and distant metastasis-free survival in patients with bilateral synchronous disease was similar to that in those with unilateral disease when controlling for subtype, stage, tumor size, grade and tumor necrosis. However, patients with bilateral synchronous clear cell renal cell carcinoma were more likely to experience local recurrence even after controlling for these covariates. The majority of patients (84%) with bilateral synchronous disease underwent bilateral surgery at a single operation. The incidence of early surgical complications was low, in that only 2 patients had urinary extravasation, 3 had acute renal failure and 1 was ultimately rendered anephric and required hemodialysis. CONCLUSIONS: The incidence of multifocality was greater in patients with bilateral synchronous renal cell carcinoma than in those with unilateral renal cell carcinoma. There were no statistically significant differences in cancer specific and distant metastasis-free survival in patients with bilateral synchronous renal cell carcinoma and unilateral renal cell carcinoma of the same histological subtype. These results suggest that subtype concordant bilateral renal cell carcinoma is a result of multiple de novo primary events rather than primary renal cell carcinoma with contralateral renal metastasis. A surgical approach is appropriate for bilateral synchronous renal cell carcinoma and most cases can be approached at a single surgical procedure with acceptable morbidity. PMID- 12629343 TI - Charlson co-morbidity index as a predictor of outcome after surgery for renal cell carcinoma with renal vein, vena cava or right atrium extension. AB - PURPOSE: Surgery is the most effective treatment for renal cell carcinoma with tumor thrombus but predictors of outcome and patient survival are variable. Co morbidity may affect therapeutic decision making and survival, although to our knowledge this factor has not been studied in patients with tumor thrombus. We analyzed the Charlson co-morbidity index as a predictor of outcome after surgery. MATERIAL AND METHODS: From 1970 to 1998, 303 patients underwent surgical resection. The Charlson index, surgical era, completeness of resection, patient age, sex, tumor level, TNM stage, grade and perinephric fat invasion were studied retrospectively as univariate and multivariate predictors of outcome. RESULTS: The level of tumor thrombus was 0 (renal vein only) in 127 patients, and I to IV in 66, 58, 36 and 16, respectively. At 5 years overall, cause specific and metastasis-free survival were 32%, 42% and 41%, while at 10 years they were 21%, 32% and 30%, respectively. For the whole cohort significant multivariate predictors of cause specific survival were metastasis (p = 0.0001), grade (p = 0.0001), perinephric fat involvement (p = 0.02) and tumor levels 0 versus I to IV (p = 0.048). The Charlson index did not predict outcome (univariate model p = 0.65). CONCLUSIONS: Characteristics of the primary tumor remained the most important predictors of cause specific survival in this cohort. The Charlson index did not predict cause specific survival in this cohort of surgically treated patients. Prospective assessment of co-morbidity in patients treated with surgery versus conservative therapy is warranted. PMID- 12629344 TI - Is there a place for radical nephrectomy in the presence of metastatic collecting duct (Bellini) carcinoma? AB - PURPOSE: We describe diagnostic and therapeutic characteristics, and long-term followup of collecting (Bellini) duct carcinoma. MATERIALS AND METHODS: Ten patients underwent surgery, including radical nephrectomy in 9 and partial nephrectomy in 1, for collecting duct carcinoma diagnosed by histological and immunohistochemical criteria. RESULTS: Collecting duct carcinoma, which represented 1.3% of renal cell carcinomas diagnosed between 1986 and 1999, developed at a mean patient age of 66.2 years (range: 50.7 to 81). It was detected clinically but never because of macroscopic hematuria. Mean tumor size was 94 mm. (range 30 to 150). Stage was pT3, pT2 and pT1 in 7, 1 and 2 cases, respectively. Seven patients had lymph node or extranodal metastases, and 8 and 2 had Fuhrman grades 3 and 4 disease, respectively. Eight patients died, including 3 early during the perioperative or postoperative period, which was attributable to major surgical difficulties associated with these invasive tumors. Two of the 3 patients who did not have metastases were alive 99 and 100 months after surgery, respectively. Median overall survival +/- SD was 9 +/- 6 and 6 +/- 8 months for patients with metastases. The all survival rate was 20% at 2 years. CONCLUSIONS: Most collecting duct carcinomas are already metastatic at presentation. Because the prognosis is dismal despite radical nephrectomy, biopsy should be performed first when radiological findings are suggestive of collecting duct carcinoma. For metastatic collecting duct carcinoma radical nephrectomy alone does not seem to be useful except for palliative reasons or in the framework of new multicentric chemotherapy protocols in progress. PMID- 12629345 TI - Reproducibility and prognostic variability of grade and lamina propria invasion in stages Ta, T1 urothelial carcinoma of the bladder. AB - PURPOSE: We assessed the reproducibility and prognostic variability of grade and lamina propria invasion in stages Ta, T1 urothelial carcinoma of the bladder. MATERIALS AND METHODS: A total of 130 consecutive stages Ta, T1 urothelial carcinomas routinely diagnosed by 15 pathologists (original diagnosis) were reviewed by 3 independent experienced pathologists using 1999 WHO criteria (diagnoses 1 to 3 and reviewer consensus diagnosis). Interreviewer disagreement cases were blindly reviewed again. Each remaining disagreement case was discussed in a multihead microscope session to attempt to solve remaining disagreements. In cases of continuing disagreement the majority diagnosis on stage and grade was considered the consensus diagnosis. Stage progression at followup was the dependent variable. Stage progression-free Kaplan-Meier survival curves and hazard ratios of each stage and grade diagnosis were calculated and prognostic variability was determined. RESULTS: There was complete interobserver agreement on stage and grade among reviewers in 80% and 59% of cases, while it was 87.7% and 75.4%, respectively, after the second review. More than 1 grade difference occurred in 1.5% of cases (0% after the second review). The consensus and original diagnoses agreed on stage and grade in 68.5% and 62.3% of cases, respectively. Assignment of individual cases to 1 category of the 1999 WHO classification per reviewer varied considerably. The incidence of cases classified as stage T1 grade 3 by the reviewers was between 12.3% and 18.9% (average 14.1%). Consensus diagnosis grade had the strongest prognostic value (HR 68.8, range 8.9 to 528.0). Of the 63 original diagnoses of stage T1 tumors the consensus diagnosis down staged 35 (55.6%) to Ta and up staged 8 (12.7%) to T2-3. Progression was more common in the 20 consensus diagnosis stage T1 cases (5 or 25%) than in the 55 original diagnosis stage T1 cases (11 or 20%). Original diagnosis stage T1 tumors that were down staged by the consensus diagnosis showed less progression than consensus diagnosis confirmed stage T1 tumors (17% versus 25%). The prognostically worst subgroup (T1 grade 3) also showed considerable prognostic variation among reviewers (28% to 76% at 5 years of followup), in that the consensus diagnosis again had the highest prognostic significance (HR 3.5, range 1.2 to 10.2). At the end of the study all pathologists expressed that they were regularly uncertain about stage and grade assessment in an individual case in a considerable percent of all cases. CONCLUSIONS: Observer prognostic variability in staging and grading is considerable with potentially strong implications for patients. Interobserver variation did not decrease using the new 1999 WHO grading classification. PMID- 12629347 TI - Outcome in patients with seminal vesicle invasion after radical cystectomy. AB - PURPOSE: We evaluated the impact of seminal vesicle invasion by transitional cell carcinoma of the bladder in a large cystectomy series. MATERIALS AND METHODS: Between January 1985 and February 2002, 1,125 cystectomies were performed at our 2 institutions. In 68 male patients there was pathologically proved tumor extension to the perivesical fat, prostatic stroma and/or seminal vesicles, including group 1: 38 to the prostatic stroma alone, group 2-12 to the seminal vesicles alone, and group 3-18 to the seminal vesicles and prostatic stroma. Complete followup was available for all patients. Overall disease specific and progression-free survival rates were calculated using the Kaplan-Maier-Method. Survival rates were compared using the log rank test. RESULTS: The overall 5-year survival rate for all 68 patients was 23.1%. The 5-year disease specific survival rates were 41.1%, 0% and 0%, and the 5-year progression-free survival rates were 32.1%, 0% and 0% for groups 1 to 3, respectively. Survival was significantly decreased in patients with seminal vesicle infiltration with or without prostatic stromal infiltration compared with prostatic involvement alone. This difference was independent of lymph node status in groups 1 versus 2 and 3. CONCLUSIONS: Seminal vesicle invasion by bladder carcinoma has a significant impact on disease specific and progression-free survival compared with prostatic stromal involvement alone. PMID- 12629346 TI - Multimodality management of urachal carcinoma: the M. D. Anderson Cancer Center experience. AB - PURPOSE: Enteric type adenocarcinomas arising in the dome of the bladder or along the urachal ligament are uncommon. To improve our understanding of urachal carcinoma and define outcome with current management, we performed a retrospective review of cases seen at the M. D. Anderson Cancer Center. MATERIALS AND METHODS: We reviewed the records of 42 patients with urachal carcinoma evaluated at our institution from 1985 to 2001. Specifically, we sought to evaluate the importance of extent of disease, surgical characteristics and systemic therapy on clinical outcome. RESULTS: Of the 42 patients 7 had clinically evident metastases at diagnosis and 35 had resectable disease that was managed initially with surgery. Overall survival from diagnosis for all 42 patients was 46 months with 40% surviving at 5 years. Of the resected cases 16 (46%) remain disease-free (median followup 31 months). Covariates associated with long-term survival were negative surgical margins (p = 0.004) and absence of nodal involvement (p = 0.01). Median survival from recognition of metastatic disease was 24 months in 26 patients in whom metastases ultimately developed. Chemotherapy for metastatic disease produced only 4 significant responses, including 3 of 9 patients treated with 5-fluorouracil and cisplatin containing regimens. CONCLUSIONS: Urachal carcinomas are usually locally advanced at presentation with a high risk of distant metastases. However, long-term survival following radical resection occurs in a significant fraction of patients (16 of 35 in our series), supporting an attempt at margin-negative, en bloc resection if at all possible. Chemotherapy appropriate for enteric type adenocarcinoma can induce objective responses but meaningful improvement in survival is not yet demonstrated. PMID- 12629348 TI - Disseminated tumor cells in bone marrow of patients with transitional cell carcinoma: immunocytochemical detection and correlation with established prognostic indicators. AB - PURPOSE: Previous investigations have demonstrated the prognostic value of disseminated cytokeratin positive cells in bone marrow of patients with breast, gastric, colon and prostate cancer. We evaluated the potential of an immunocytochemical assay, using a monoclonal antibody against cytokeratin 18 (CK 18), for the detection of disseminated tumor cells in bone marrow aspirates of patients with transitional cell carcinoma. MATERIALS AND METHODS: Bone marrow aspiration was performed preoperatively on 128 patients with transitional cell carcinoma of various stages and on 27 controls with nonmalignant disease. Cytospin preparations of mononuclear bone marrow cells were incubated with a monoclonal anti-CK 18 antibody and stained using the alkaline phosphatase anti alkaline phosphatase technique. RESULTS: Of the patients with transitional cell carcinoma 29.7% and none of the controls had a CK 18 positive bone marrow result. A significant correlation between the incidence of CK 18 positive cells in bone marrow and invasive transitional cell carcinoma (p <0.01), lymph node involvement (p <0.01), medium/high grade transitional cell carcinoma (p <0.01) and tumor progression in recurrent transitional cell carcinoma (p <0.05) was demonstrated. Furthermore, the mean number of CK 18 positive cells in bone marrow aspirates of patients with stage M+ and/or N+ disease was nearly 3 times as high as that of patients without clinically evident metastatic disease (10.4 versus 3.8 CK 18 positive cells per patient). CONCLUSIONS: A significant correlation between the incidence of CK 18 positive bone marrow results in patients with transitional cell carcinoma and established risk factors could be demonstrated in our study. Further prospective followup studies should be performed to determine the prognostic value of these findings. PMID- 12629349 TI - Leptin influences cellular differentiation and progression in prostate cancer. AB - PURPOSE: Several studies have shown a positive association of dietary fat with prostate cancer. Leptin, a peptide hormone that has a role in the regulation of body weight, currently serves as a more accurate biomarker for total body fat. We designed a study to determine whether leptin influences cellular differentiation and the progression of prostate cancer. MATERIALS AND METHODS: In this study we investigated serum leptin in 21 patients with prostate cancer, 50 with benign prostatic obstruction and 50 healthy individuals matched for sex, body mass index and age. Patients with cancer were stratified into 2 groups by the disease spread, including groups 1--organ confined and 2--advanced disease, and into 3 groups by the differentiation degree, including groups 3--Gleason sum 2 to 4 or well differentiated, 4--Gleason sum 5 to 7 or moderately differentiated and 5- Gleason sum 8 to 10 or poorly differentiated. RESULTS: We noted significant differences in serum leptin in the cancer versus control and cancer versus benign prostatic obstruction groups. In addition, in the prostate cancer group serum leptin correlated with prostate specific antigen and biopsy Gleason score. We also observed significant differences in serum leptin in groups 1 versus 2, 3 versus 5 and 4 versus 5. CONCLUSION: Leptin may have roles in the development of prostate cancer through testosterone and factors related to obesity. It influences cellular differentiation and the progression of prostate cancer. PMID- 12629350 TI - Associations of serum testosterone with microvessel density, androgen receptor density and androgen receptor gene polymorphism in prostate cancer. AB - PURPOSE: We investigate potential associations of serum testosterone with microvessel density, androgen receptor expression and AR gene polymorphism in men with untreated prostate cancer. MATERIAL AND METHODS: Serum luteinizing hormone, follicle-stimulating hormone, estradiol and testosterone were determined in men with newly diagnosed prostate cancer. The number of tumor vessels per 0.46 mm. and androgen receptor density (as the percent positive nuclei) were quantified immunohistochemically on prostate cancer areas of prostate biopsy specimens. Polymorphisms within the AR gene (number of CAG repeats) were determined by polymerase chain reaction and restriction fragment length polymorphism analysis using DNA from peripheral blood. RESULTS: The 39 men entered into this study were grouped into 16 with low (3 ng./ml. or less, group 1) and 23 with normal (greater than 3 ng./ml., group 2) serum testosterone. Mean prostate specific antigen +/- SD was significantly lower in group 1 than in group 2 (18.8 +/- 11.1 versus 27.2 +/- 12.2 ng./ml., p = 0.03). Mean Gleason score (7.4 +/- 1.3 versus 6.0 +/- 1.2, p = 0.01), androgen receptor density (96.6% +/- 2.8% versus 84.8% +/- 7.2%, p = 0.03) and tumor vessel density (63.0 +/- 30.8/0.46 versus 39.0 +/- 22.9/0.46 mm.2, p = 0.007) were significantly higher in group 1 than in group 2. The number of CAG repeats within the AR gene did not correlation with serum androgen. CONCLUSIONS: Low serum testosterone in men with newly diagnosed prostate cancer is associated with higher tumor microvessel and androgen receptor density as well as with higher Gleason score, suggesting enhanced malignant potential. PMID- 12629351 TI - Hepsin and maspin are inversely expressed in laser capture microdissectioned prostate cancer. AB - PURPOSE: Recent studies have shown that hepsin, a serine protease, is over expressed in prostate cancers, implicating hepsin activity in tumor invasion. Using microarray technology we have previously identified 22 genes that were up regulated in high grade prostate cancers compared with benign prostatic hyperplasia. Of them hepsin was the most differentially over expressed. In the current report we compare hepsin to maspin (BD Transduction Laboratories, San Diego, California), a serine protease inhibitor (serpin), to measure the balance between levels of serine proteases and serpins, which are considered to be a critical determinant of net proteolytic activity. MATERIALS AND METHODS: We combined the technique of laser capture microdissection with gene expression monitoring by micro-array analysis to investigate the gene expression profiles of prostate cells of different histological types. We also studied maspin immunohistochemically. RESULTS: We observed that hepsin as well as 7 of 22 previously reported up-regulated genes demonstrated a pattern of increasing expression with increasing malignant phenotype. In contrast, the expression of maspin (a serpin) decreased with increasing malignancy of prostate cancers. Using immunohistochemistry we observed that maspin protein is expressed strongly in benign prostatic tissues and slightly in grade 3 prostate cancers, and is absent in grade 4/5 cancers. CONCLUSIONS: We conclude that the increased ratio of hepsin to-maspin may have an important role in prostate cancer progression and invasion. PMID- 12629352 TI - Pretreatment predictors of time to cancer specific death after prostate specific antigen failure. AB - PURPOSE: Whether pretreatment factors that predict for time to prostate specific antigen (PSA) failure also predict for time to prostate cancer specific death after PSA failure for patients with competing causes of mortality treated during the PSA era was the subject of this study. MATERIALS AND METHODS: Of 415 men with a median age of 73 years who underwent external beam radiation therapy between 1988 and 2001 for clinically localized prostate cancer 160 (39%) experienced PSA failure and 96 (23%) died. In 46 men (48%) the cause of death was prostate cancer. Cox regression multivariable analyses (multivariable analysis) were performed to evaluate the ability of the pretreatment PSA and centrally reviewed biopsy Gleason score to predict time to prostate cancer specific death after PSA failure. RESULTS: When analyzed as categorical variables using multivariable analysis, biopsy Gleason score 4 + 3 (p = 0.02), 8 to 10 (p = 0.02) disease and a pretreatment PSA greater than 20 ng./ml. (p = 0.03) were significant predictors of time to prostate cancer specific death after PSA failure. Estimates of prostate cancer specific death 5 years after PSA failure were 24%, 40% and 59% (p = 0.01) for patients with a biopsy Gleason score < or = 6, 3 + 4, 4 + 3 or higher and 22%, 40% and 60% (p = 0.04) for patients with a pretreatment PSA of 10 or less, greater than 10 and 20 or less, or greater than 20 ng./ml., respectively. CONCLUSIONS: Patients at high risk for PSA failure after radiation therapy based on pretreatment PSA greater than 20 ng./ml. or biopsy Gleason score 4 + 3 or greater are also at high risk for death from prostate cancer after PSA failure despite competing causes of mortality. PMID- 12629353 TI - Preoperative p27 status is an independent predictor of prostate specific antigen failure following radical prostatectomy. AB - PURPOSE: p27 is an important cell cycle regulator, and decreased expression in radical prostatectomy specimens is associated with an increased risk of prostate specific antigen (PSA) failure. To our knowledge no prior study has shown that preoperative p27 status independently predicts recurrence after radical prostatectomy. MATERIALS AND METHODS: The prostate needle biopsy specimens of 161 men treated with radical prostatectomy were examined for p27 expression using immunohistochemistry. Various p27 cut points were examined for their ability to separate patients into groups with different risk for time to biochemical recurrence following radical prostatectomy. The best p27 cut point was compared to other clinical variables (PSA, clinical stage, age, biopsy Gleason score and percent of prostate needle biopsy with cancer) on multivariate analysis to determine which variables independently predicted biochemical failure. RESULTS: A p27 cut point of less than 45% positive staining cells resulted in significant preoperative risk stratification for time to PSA failure (HR 2.41, p = 0.010). On multivariate analysis serum PSA (HR 1.04, p = 0.011), biopsy Gleason score (HR 1.51, p = 0.011), percent of biopsy tissue with cancer (HR 10.01, p = 0.001) and less than 45% p27 positive cells (HR 2.44, p = 0.014) were all independent predictors of biochemical recurrence. CONCLUSIONS: Preoperative p27 expression is an independent predictor of time to biochemical recurrence following radical prostatectomy. Patients with less than 45% p27 positive cells in the prostate needle biopsy specimen have almost a 2.5-fold increased risk of biochemical recurrence. To our knowledge this study is the first to show that p27 status of the prostate needle biopsy specimen can be used before radical prostatectomy to predict biochemical failure. PMID- 12629354 TI - Prostate cancer with large glands treated with 3-dimensional computerized tomography guided pararectal brachytherapy: up to 8 years of followup. AB - PURPOSE: We report post-brachytherapy results in patients with cancer in a large prostate. MATERIALS AND METHODS: From June 1, 1994 to June 30, 2000, 331 consecutive patients with a large prostate of 50 to 180 cm.3 (median 69) were treated with 3-dimensional computerized tomography guided brachytherapy. Patient age was 42 to 90 years (median 69). Of these patients 327 were available for followup for 2 to 8 years (median 4.5). Patients were stratified according to risk profile. The high risk group had 1 or more high risk factors (prostate specific antigen [PSA] greater than 20 ng./ml., Gleason greater than 7, stage T2b, T3a or T3b) or 2 intermediate risk factors (PSA 10 to 20 ng./ml. and Gleason 7). The high risk group was further stratified into subgroups with a similar risk profile. The intermediate risk group had only 1 high risk factor (PSA 10 to 20 ng./ml. or Gleason 7). The low risk group had PSA less than 10 ng./ml., Gleason less than 7 and stage T1a, b, c or T2a. A dose of 144 Gy. with 125I or 120 Gy. with 103Pd was achieved in 90% to 100% of the target. A total of 31 patients (9%) had previously undergone transurethral resection and 198 (60%) were treated with 3 months of neoadjuvant androgen ablation. RESULTS: Biochemical disease-free survival was achieved in 90% of the 182 patients at high risk, 96% of the 52 at intermediate risk and 99% of the 93 at low risk. Seven patients (2%) required catheterization during year 1 for urinary retention, 11 (3%) required transurethral prostate resection 1 to 4 years after implantation, 3 patients (1%) had grade 1 or 2 incontinence after repeat transurethral prostate resection and 4 (1%) had grade 3 or 4 rectal complications. CONCLUSIONS: The 3-dimensional computerized tomography guided pararectal permanent implant results in a high level of biochemical control with low morbidity at 2 to 8 years in patients with prostate cancer who have a large prostate. There was less favorable biochemical control in patients with PSA greater than 20 ng./ml., Gleason 7 or greater and seminal vesicle invasion. PMID- 12629355 TI - Value of [11C]choline-positron emission tomography for re-staging prostate cancer: a comparison with [18F]fluorodeoxyglucose-positron emission tomography. AB - PURPOSE: We compared [11C]choline-positron emission tomography (PET) with [18F]fluorodeoxyglucose-PET for re-staging prostate cancer in a group of 100 patients. MATERIALS AND METHODS: A total of 100 consecutive patients referred for whole body [18F]fluorodeoxyglucose-PET for clinical prostate re-staging after radical treatment for prostate cancer were retrospectively included in the study. Mean prostate specific antigen (PSA) was 6.57 ng./ml. In all cases [11C]choline PET was also performed. PET studies were done with a multiring device 5 minutes after intravenous injection of approximately 370 MBq. [11C]choline and 60 minutes after injection of approximately 370 MBq. [18F]fluorodeoxyglucose. PET findings were compared with those obtained with different conventional imaging and with PSA assessed at the time of PET and 1 year later. RESULTS: Areas of abnormal focal increases were noted in 47% of patients on [11C]choline-PET and in 27% on [18F]fluorodeoxyglucose-PET. Of the 100 patients 49 had positive conventional imaging findings. All except 14 [11C]choline-PET findings were concordant with conventional imaging, including 6 negative and 8 positive conventional imaging results. All except 1 [11C]choline-PET negative cases also had negative conventional imaging after 1 year. PSA at 1 year remained stable or decreased in 80% and 62% of [11C]choline-PET negative and positive cases, respectively. CONCLUSIONS: [11C]choline-PET seems to be useful for re-staging prostatectomy cases with increasing serum PSA levels. It is superior to [18F]fluorodeoxyglucose PET and complementary to conventional imaging but with the advantage of staging disease at a single step. PMID- 12629356 TI - Investigation of the safety and accuracy of intraoperative gamma probe directed biopsy of bone scan detected rib abnormalities in prostatic adenocarcinoma. AB - PURPOSE: We evaluated the technique of intraoperative gamma probe directed rib biopsy in patients with suspected metastatic prostate adenocarcinoma. This technique can be used to identify accurately the rib in question, reliably obtain sufficient tissue for diagnosis, be performed with minimal patient morbidity and potentially alter the course of therapy. MATERIALS AND METHODS: From 1996 to 2001, 8 patients with biopsy proved adenocarcinoma of the prostate and suspicious rib lesions on radionuclide bone scanning underwent open rib biopsy as part of the evaluation for metastatic disease. Mean prostate specific antigen in the patient population was 17.1 ng/ml (range 6.1 to 36.5) and clinical stage was T1c to T3c. A new technique of intraoperative gamma probe directed biopsy was used to localize and resect the rib in question. At 6 to 12 hours before the operation each patient received an intravenous injection of 28 mCi. (99m)technetium oxidronate. The hand held, pencil sized gamma probe in a sterile sleeve was used to localize the area of greatest activity in the target bone and 3 cm. of bone were resected. RESULTS: Of the 8 patients who underwent the procedure 2 had metastatic prostate cancer on final rib pathological findings. Four of the remaining 5 patients had benign rib lesions (an old rib fracture) and 1 had metastatic lung cancer. The hot spot on bone scan was localized with 100% accuracy using our technique and a pathological diagnosis was made in all cases. Mean operative time was 61 minutes and estimated blood loss was less than 20 ml. in all cases. Seven of the 8 patients were discharged home the same day, while 1 required overnight hospitalization. There was 1 intraoperative complication of inadvertent entry into the pleural cavity, resulting in a small pneumothorax, which was treated with small chest catheter drainage and observation. CONCLUSIONS: Intraoperative gamma probe directed rib biopsy of suspected metastatic lesions in patients with prostate cancer can be safely and accurately performed with minimal patient morbidity. The information obtained using this technique can be used to tailor treatment decisions for this subset of patients with prostate cancer. PMID- 12629357 TI - Second primary cancers in men with prostate cancer: an increased risk of male breast cancer. AB - PURPOSE: Evaluation of second primary cancers provides valuable insight about etiology and shared risk factors. Studies of second primary cancers following prostate cancer conclude that overall risk of second primary cancers decreases. However, risk of bladder cancer and kidney cancer increases. We examine the risk of common and rare second primary cancers following prostate cancer in a large population based cohort to identify possible common etiological factors. MATERIALS AND METHODS: All prostate cancer cases in the Swedish Cancer Registry (135, 713) from 1958 to the end of 1996 constituted the study base. Risk (standardized incidence ratio) of second primary cancers was calculated as the ratio between observed and expected number of cancers. We used 2-tailed 95% confidence intervals (CI) to test significance. RESULTS: An overall increased risk (standardized incidence ratio 1.17, 95% CI 1.15-1.19) of second primary cancers was found but was only seen in the first 6 months of followup (ratio 3.45, 3.32-3.57). The most interesting finding was an increased risk (ratio 2.01, 95% CI 1.44-2.74) of male breast cancer. Other tumor sites with increased risk were the small intestine (standardized incidence ratio 1.39, 95% CI 1.09-1.51), skin melanoma (ratio 1.33, 95% CI 1.16-1.51) and endocrine tumors (ratio 1.41, 95% CI 1.13-1.74). CONCLUSIONS: A small but increased risk of second primary cancers following prostate cancer was found, most likely due to increased surveillance during the first 6 months after diagnosis. However, following prostate cancer there is an increased risk of endocrine related second primary cancers such as male breast cancer and carcinoids in the small intestine. To our knowledge these associations have not been reported previously, and they warrant more study. PMID- 12629358 TI - Modified radical inguinal lymphadenectomy for carcinoma of the penis: technique and results. AB - PURPOSE: Modified radical inguinal lymphadenectomy for carcinoma of the penis is presented that satisfies the requirement for complete groin dissection, while significantly decreasing postoperative complications. MATERIALS AND METHODS: Eight patients with squamous cell carcinoma and 2 with leiomyosarcoma of the penis underwent bilateral modified inguinal lymphadenectomy, including removal of the superficial and deep inguinal lymph nodes. To avoid damage to the vessels of the groin region that run parallel to the inguinal ligament and lie in the fat of the superficial layer of the superficial fascia dissection is done beneath this layer. The proper cleavage plane is just above the membranous layer of the superficial fascia, beneath which the superficial inguinal lymph nodes are located. The saphenous vein is preserved and the sartorius muscle is left in situ, so as not to disturb collateral lymphatic drainage. RESULTS: At a followup of 6 to 104 months no skin necrosis, infection or deep venous thrombosis occurred. In 2 patients early moderate lymphedema of the lower extremities resolved with time, 2 had scrotal edema and 3 had a transient lymphocele. CONCLUSIONS: As described, modified radical inguinal lymphadenectomy decreases the morbidity associated with groin dissection, while removing superficial and deep inguinal lymph nodes. PMID- 12629359 TI - Repeat retroperitoneal lymph node dissection for metastatic testis cancer. AB - PURPOSE: We report pathological results, perioperative complications and patient outcome in 21 men after repeat retroperitoneal lymph node dissection for metastatic testis cancer. MATERIALS AND METHODS: We reviewed an institutional tumor registry at our cancer center and identified 417 patients who underwent retroperitoneal lymph node dissection for testis cancer during a 21-year period. Of these 417 patients 21 underwent repeat retroperitoneal lymph node dissection. We reviewed preoperative patient characteristics, operative data and pathological findings from repeat lymphadenectomy, and determined patient disease status, morbidity and mortality after surgery. RESULTS: We identified viable germ cell tumor in 5 patients (24%), teratoma in 14 (67%) and fibrosis or necrosis only in 5 (24%). Intraoperatively subadventitial dissection of the aorta occurred in 2 cases, which was severe enough in 1 to require an aortic graft. The most common postoperative complications were prolonged ileus or partial bowel obstruction and chylous ascites in 6 and 3 patients, respectively. Six patients died, including 5 of disease progression and 1 of postoperative pulmonary embolus. At a mean followup of 4.7 years (range 0.1 to 14) 15 patients (71%) were alive and 14 (67%) were disease-free. CONCLUSIONS: Repeat retroperitoneal lymph node dissection is safe and effective in the majority of patients with recurrent or residual retroperitoneal masses after initial multimodality treatments for metastatic testis cancer. Overall perioperative morbidity and mortality are low and yet the potential for significant vascular complications warrants careful preoperative planning and intraoperative judgment. PMID- 12629360 TI - Surgical management of urogenital injuries at a war hospital in Bosnia Hrzegovina, 1992 to 1995. AB - PURPOSE: Our experience with urogenital surgery during the Bosnia-Herzegovina conflict of 1992 to 1995 is compared with data from previous wars and peacetime practice. MATERIALS AND METHODS: A total of 5,370 wounded patients, including 136 (2.5%) with 1 or more urogenital injuries, were treated at a war hospital in northern Bosnia-Herzegovina during 37 months. The hospital was based at a village school and was poorly equipped but it was only 10 to 16 km. from the front line and average transportation time was 30 minutes. RESULTS: Of the 136 patients 72 (52.9%) were injured by explosive weapons, while the other 64 (47.1%) had bullet wounds. The 169 urogenital injuries were to the kidney in 65 cases (38.5%), scrotum in 44 (26%), bladder in 23 (13.6%), penis in 16 (9.5%), urethra in 9 (5.3%), ureter in 6 (3.6%) and other in 6. There was associated damage to organs other than the urogenital system in 116 patients (85.3%). Preference was given to organ sparing operations when possible, but 33.8% of renal injuries required nephrectomy and orchidectomy was performed for 58.3% of testicular injuries. CONCLUSION: In war settings when injuries are often severe and multiple, and the hospital may lack staff, instruments and other medical supplies, the surgeon frequently must improvise. Even so, the results achieved need not fall far below those of peacetime surgery provided that, as in this hospital in Bosnia Herzegovina, wounded patients present to the surgeon rapidly, they are young and the surgeons are experienced with the management of war injury. PMID- 12629361 TI - Fibrin sealant for the management of genitourinary injuries, fistulas and surgical complications. AB - PURPOSE: We report our preliminary experience with the use of fibrin sealant to manage iatrogenic urinary tract injuries, complex urinary fistulas, and urological surgical complications. MATERIALS AND METHODS: Topical fibrin sealant was used in 19 patients for iatrogenic urinary tract injury during gynecological or general surgical procedures (7), complex urinary fistulas (5) or urological surgical complications (7). RESULTS: Successful resolution of the injury, fistula or complication was attained after a single application of fibrin sealant in the 18 patients (94.7%) in whom a direct injection technique was used. The only failure (formation of a vesicovaginal fistula) occurred with the air driven sprayed sealant delivery technique after sutured closure of iatrogenic cystotomy during vaginal hysterectomy. CONCLUSIONS: Liquid fibrin sealant appears to be safe and prudent for use in urological "damage control" from trauma, fistulas or surgical complications. Direct injection over a sutured urinary anastomosis appears to be highly effective in preventing urinary drainage. Additional study is needed to define further the role and best use of tissue adhesives in urology. PMID- 12629362 TI - Expanded experience with laparoscopic nephrectomy and autotransplantation for severe ureteral injury. AB - PURPOSE: Complex injuries of the upper ureter with significant loss of length are difficult to manage. Traditional options include nephrectomy, bowel interposition and autotransplantation. The advent of laparoscopic donor nephrectomy has created a new option when preservation of functioning renal parenchyma is desirable. We update our experience with severe iatrogenic ureteral injuries requiring laparoscopic nephrectomy and autotransplantation. MATERIALS AND METHODS: Seven patients with severe ureteral injuries were referred for definitive management. In all cases preoperative evaluation demonstrated comparable split renal function and anatomy not amenable to routine reconstruction. Transperitoneal laparoscopic nephrectomy and autotransplantation to the ipsilateral pelvis were attempted. RESULTS: Of the patients 5 had a history of nephrolithiasis with intervention resulting in the ureteral injury and 5 underwent prior endoscopic or open management of the defects. All cases were associated with dense perinephric and perihilar fibrosis. Operative and warm ischemic times averaged 508 and 5 minutes, respectively. After ex vivo graft preparation, ureteral and vessel length and quality were adequate for transplantation in 6 cases. In the other patient the renal pelvis was not amenable for reconstruction. Urinary drainage consisted of ureteroneocystostomy in 2 cases and ipsilateral ureteroureterostomy in 4. There were no intraoperative complications and all grafts functioned immediately. Hospitalization averaged 6.4 days. At a mean followup of 17 months imaging studies demonstrated functioning renal autographs with a mean decrease in serum creatinine of 5%. CONCLUSIONS: Laparoscopic nephrectomy in conjunction with autotransplantation is a viable option for severe proximal ureteral loss. Acceptable perioperative morbidity and excellent graft function were obtained in all cases when the kidney was transplanted. Close cooperation with a transplant team is crucial to coordinate graft harvest, ensure appropriate organ preparation and select the optimal urinary anastomosis. PMID- 12629363 TI - Editorial: new technology in renal trauma. PMID- 12629364 TI - Sacral neuromodulation for the symptomatic treatment of refractory interstitial cystitis: a prospective study. AB - PURPOSE: The efficacy of sacral neuromodulation for the treatment of symptoms in patients with refractory interstitial cystitis was evaluated. MATERIALS AND METHODS: A total of 25 patients with a mean age of 47 years and refractory interstitial cystitis were prospectively evaluated with a trial of sacral nerve stimulation. Patients who demonstrated 50% improvement in frequency, nocturia, voided volume and average pain qualified for permanent sacral nerve stimulator implantation. Treatment success was measured by voiding diary, report of average pain, and response to the Interstitial Cystitis Symptom Index and Interstitial Cystitis Problem Index. RESULTS: Of the 25 patients 17 qualified for permanent sacral nerve stimulator implantation. At an average of 14 months followup mean daytime frequency and nocturia improved from 17.1 to 8.7 and 4.5 to 1.1, respectively (p <0.01). Mean voided volume increased from 111 to 264 ml. (p <0.01). Report of average pain decreased from 5.8 to 1.6 points on a scale of 0 to 10 (p <0.01). Interstitial Cystitis Symptom and Problem Index scores decreased from 16.5 to 6.8 and 14.5 to 5.4, respectively (p <0.01). Of the 17 patients 16 (94%) with a permanent stimulator demonstrated sustained improvement in all parameters at the last postoperative visit. CONCLUSIONS: Results of this prospective clinical study demonstrate that sacral neuromodulation is a safe and effective treatment for the dysfunctional voiding and pelvic pain in patients with interstitial cystitis who are refractory to other forms of treatment. PMID- 12629365 TI - A prospective controlled quantitative study of ultrastructural changes in the underactive detrusor. AB - PURPOSE: We quantify and compare ultrastructural changes seen at high power on electron microscopic examination of detrusor biopsies from underactive and control bladders. MATERIALS AND METHODS: The detrusor underactivity group consisted of 14 patients (mean age 64.0 years) with post-micturition residuals greater than 300 ml., impaired contractility and no obstruction on urodynamics. A total of 17 patients who voided normally voiders with residual volume less than 50 ml. served as the control group. Bladder biopsies were obtained endoscopically using cold cup technique and processed using standard methods. All specimens were randomized and studied at high power (x12,000 to x24,000) by an examiner who was blinded to the groups. All complete cells within each random grid field were counted up to a total of 500. The number of disrupted cells per 500 cells was noted. RESULTS: Disruptive cell profiles were found in all biopsies. Median cell count was 20 cells/500 in 14 controls and 96.5 cells/500 in the underactive detrusor group (p <0.001). The number of disruptive cell profiles did not correlate with age in the control group (r = 0.34, p = 0.18). CONCLUSIONS: There were distinct morphological changes in the detrusor associated with bladder dysfunction with approximately 4 times more disruptive cells in patients with an underactive bladder. However, there was no correlation between age and number of disruptive cells, suggesting that this is not a process of aging per se. Ultrastructure studies may have a role as an adjunct to urodynamics in the diagnosis of bladder dysfunction. PMID- 12629366 TI - Free neurovascular transfer of latisstmus dorsi muscle for the treatment of bladder acontractility: II. Clinical results. AB - PURPOSE: Until now patients with bladder acontractility were destined to lifelong clean intermittent catheterization with all of its inherent risks. Previous experimental studies demonstrated that voluntary voiding can be restored by microneurovascular free transfer of a carefully selected muscle flap. We present the selection criteria, modifications in technique, followup schedule and long term results in 20 patients treated with transplantation of latissimus dorsi muscle to the bladder. MATERIALS AND METHODS: In 20 patients with bladder acontractility requiring intermittent catheterization for at least 2 years we performed latissimus dorsi detrusor myoplasty. Preoperative evaluation included urodynamic assessment, cystoscopy, upper tract imaging and electromyography of the rectus muscle. The procedure involves transfer of a free neurovascular latissimus dorsi muscle flap to the pelvis where it is anastomosed to the lowest motor branches of the intercostal nerve and deep inferior epigastric vessels. Patients were instructed to attempt voluntary voiding 3 months postoperatively. Followup included urodynamic evaluation, biannual Doppler ultrasonography and annual dynamic computerized tomography. RESULTS: Annual dynamic computerized tomography and/or biannual Doppler ultrasonography demonstrated vascularization and contractility of all transplanted muscle flaps. Mean followup is 44 months (range 18 to 74). Of the 20 patients 14 were able to void spontaneously within 4 months postoperatively with post-void residual volumes of less than 100 cc, voluntary voiding was restored by bladder neck incision in 4 and 2 (10%) still require self-catheterization. Postoperative detrusor pressures ranged from 5 to 218 cm. H2O (mean 72, median 55). None of the patients had morphological and functional changes of the upper tract, or de novo incontinence postoperatively. CONCLUSIONS: Functioning free muscle transplantation was able to restore voluntary voiding in patients who had previously been dependent on long-term catheterization. Voluntary voiding has been maintained several years postoperatively without deterioration of upper tract function. PMID- 12629367 TI - Relationship of bladder dysfunction to lesion site in multiple sclerosis. AB - PURPOSE: We investigated the relationship of voiding dysfunction type and the lesion site in patients with multiple sclerosis. MATERIALS AND METHODS: Voiding dysfunction was evaluated in 32 patients with multiple sclerosis using the International Prostate Symptom Score and urodynamic tests. Lesion sites were determined by combined neurological examination and magnetic resonance imaging findings. RESULTS: Compared with reports from Western countries the ratio of emptying-to-filling symptoms was high in Japan. Of urinary symptoms only filling correlated with disability status and disease duration. Urinary symptoms were not related to lesion sites. Urodynamic evaluation revealed detrusor hyperreflexia in 14 of 32 patients, hyporeflexia or areflexia in 12, detrusor hyperreflexia with impaired contractile function in 4, a low compliance bladder in 1 and normal function in 1. Of 14 patients with hyperreflexia 13 had overactive sphincter concurrently. Incompetent sphincter was identified in 2 patients who had detrusor hyperreflexia with impaired contractility and in 1 with a low compliance bladder. A significant correlation was noted for a pontine lesion and detrusor hyporeflexia, and for a cervical cord lesion and detrusor-sphincter dyssynergia. CONCLUSIONS: Detrusor hyporeflexia and detrusor-sphincter dyssynergia are indicative of a pontine and cervical spinal cord lesion, respectively. Thus, the lesion site in the central nervous system may be a major determinant of the type of bladder and urethral sphincter dysfunction. The high prevalence of emptying symptoms in Japanese patients may reflect the prevalence of detrusor hyporeflexia and detrusor-sphincter dyssynergia. PMID- 12629369 TI - Impact of rectal distention on the results of evaluations of lower urinary tract sensation. AB - PURPOSE: To our knowledge no systematic studies have been performed in humans to test the interaction of normal bladder and rectal sensory function. However, symptoms affecting the different pelvic viscera often coexist. MATERIALS AND METHODS: In 15 healthy female volunteers sensations of bladder filling were evaluated during 2 consecutive cystometric studies, including 1 with an empty rectum and 1 with a full rectum. Similarly the electrical perception threshold was determined in the bladder when the rectum was empty and again when the rectum was full. RESULTS: Rectal distention did not change bladder compliance during filling. When the rectum was distended, sensations of bladder filling were reported at smaller volumes. On the other hand, electrical perception thresholds in the bladder were higher when the rectum was full. CONCLUSIONS: The state of the rectum significantly influences the sensations of bladder filling and electrical bladder stimulation. These results show that before sensory testing of the bladder rectal fullness should be examined and if necessary the rectum should be emptied. The different effect of rectal distention on filling and electrical sensations represents an additional argument for different innervation of the 2 sensations in the lower urinary tract. PMID- 12629368 TI - Effects of subthalamic nucleus stimulation on urodynamic findings in patients with Parkinson's disease. AB - PURPOSE: Although the effects of subthalamic nucleus stimulation on the control of motor symptoms in patients with Parkinson's disease have been demonstrated, to our knowledge there are no data on effects of this treatment on voiding. We evaluated differences in urodynamic findings in patients with Parkinson's disease during on and off subthalamic nucleus stimulation status. MATERIALS AND METHODS: We evaluated 3 males and 2 females with Parkinson's disease. All patients had undergone surgical bilateral implantation of subthalamic nucleus electrodes 4 to 9 months before our observation. Urodynamic evaluation was performed during chronic subthalamic nucleus stimulation and 30 minutes after turning off the stimulators. Certain parameters were evaluated, including bladder compliance and capacity, first desire to void volume, bladder volume of appearance (reflex volume) and amplitude of detrusor hyperreflexic contractions, maximum flow, detrusor pressure at maximum flow and detrusor-sphincter coordination. Results were compared statistically. RESULTS: Statistically significant differences in urodynamic data obtained during on and off subthalamic nucleus stimulation status were noted. In particular bladder capacity and reflex volume were increased for on status (median 320 versus 130 ml., p = 0.043 and 250 versus 110, p = 0.043, respectively). The amplitude of detrusor hyperreflexic contractions was decreased for on status but the difference was not significant (median 23 versus 37 cm. H2O, p = 0.223). No differences were noted in the other urodynamic parameters considered during the filling and voiding phases. CONCLUSIONS: Our experience shows that subthalamic nucleus stimulation seems to be effective for decreasing detrusor hyperreflexia in Parkinson's disease cases. This finding confirms a role for basal ganglia in voiding control. PMID- 12629370 TI - Transvaginal ultrasound for the diagnosis of urethral diverticulum. AB - PURPOSE: We reviewed our experience with urethral diverticula and transvaginal ultrasound to evaluate female patients with this condition. MATERIALS AND METHODS: All female patients who underwent transvaginal ultrasound during evaluation for urethral diverticulum at our institution between May 1995 and April 2002 were identified by searching a data base. We reviewed the diagnostic evaluation and compared diagnostic techniques with transvaginal ultrasound for diagnosing urethral diverticulum. RESULTS: Of the 25 patients with suspected urethral diverticulum who underwent transvaginal ultrasound as a diagnostic procedure 10 (40%) were diagnosed with urethral diverticulum, including 10 in whom it was confirmed by surgery or other diagnostic procedures. Transvaginal ultrasound was less expensive than the other diagnostic modalities and in no case did it miss a urethral diverticulum that was identified by another diagnostic technique. Urethral diverticulum was detected on 1 of 3 voiding cystourethrograms (33%) and this study missed the diagnosis in 1 case that was diagnosed by transvaginal ultrasound. Three cases of urethral diverticulum were noted on transvaginal ultrasound after they were missed by cystoscopy. Videourodynamics were unable to diagnose urethral diverticulum. CONCLUSIONS: Transvaginal ultrasound is effective for evaluating patients with suspected urethral diverticulum. It is less expensive and may identify diverticula missed by other diagnostic modalities. PMID- 12629371 TI - Experience with urethral stent explantation. AB - PURPOSE: We examined the timing, causes and results of the explantation of UroLume (American Medical Systems, Minnetonka, Minnesota) urethral stents. MATERIALS AND METHODS: The North American Study Group enrolled 465 patients in a trial of UroLume stent placement, including 69 (14.8% of the series) who required removal of a total of 73 stents (15.6% of stents) in 7 years. The time of explantation, reason for stent removal and local histological findings were determined from patient charts and reviewed in relation to the indication for stent placement. RESULTS: In patients treated for bladder outlet obstruction secondary to benign prostatic hyperplasia 23% of the stents were removed, as were 5% of those implanted in patients with bulbar urethral stricture and 22% of those in patients with detrusor-sphincter dyssynergia (see figure). Of the explantations 43.8% were done during year 1. Migration and/or inappropriate placement was the cause in 38.4% of cases. The most common stent site tissue finding was focal inflammation with a hyperplastic tissue response. Stent specific malignant changes were not observed. Stent removal was feasible with no sequelae. CONCLUSIONS When used appropriately, the UroLume endoprosthesis has a low incidence of failure. Stent removal is technically feasible and options are available for subsequent therapy. Local tissue reaction is minimal. PMID- 12629372 TI - A randomized, placebo controlled, multicenter study to evaluate the safety and efficacy of rofecoxib in the treatment of chronic nonbacterial prostatitis. AB - PURPOSE: We determine the effects of treatment with rofecoxib and placebo in patients with chronic prostatitis. MATERIALS AND METHODS: Patients diagnosed with chronic nonbacterial prostatitis were randomized to 6 weeks of 25 or 50 mg., rofecoxib or placebo in a double-blind multicenter study with a 1-week run in of placebo. End points included the National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI) (average pain score item 4 primary end point), and patient global assessment questions of pain, disease activity and response to therapy. RESULTS: A total of 161 patients were randomized in the study. The NIH-CPSI total, domain and pain scores significantly decreased from baseline in all groups and, although the mean scores numerically favored the rofecoxib groups, the difference was not significantly different among groups. There was a trend for the percentage of patients with a 25% (or 6 point) improvement in total score being superior on rofecoxib versus placebo with the difference being significantly different (p <0.05) for the 50 mg. rofecoxib group. Patient global assessment of pain, response to therapy and disease activity also favored rofecoxib over placebo (p <0.05, p = 0.07, p = 0.06, respectively). Of the patients 79% on 50 mg. rofecoxib versus 59% on placebo reported no or mild pain, and 56% of patients on 50 mg. rofecoxib versus 27% on placebo experienced significant improvement in quality of life (p <0.005). Rofecoxib was generally well tolerated. CONCLUSIONS: To our knowledge this study is the first to evaluate rofecoxib versus placebo in patients with prostatitis and the first large multicenter treatment study to use the NIH-CPSI. Subjective assessment with patient global questions may be more sensitive to change than the NIH-CPSI and, therefore, may be a better tool to use in future therapeutic trials. Although 6 weeks of rofecoxib treatment appear to benefit many men diagnosed with chronic prostatitis/chronic pelvic pain syndrome further studies are needed. PMID- 12629373 TI - Long-term results of multimodal therapy for chronic prostatitis/chronic pelvic pain syndrome. AB - PURPOSE: Chronic prostatitis/chronic pelvic pain syndrome is a prevalent and multifactorial condition. Many patients have the condition for years despite conventional therapies. We assess the outcomes of multimodal therapy in patients with long-standing chronic prostatitis/chronic pelvic pain syndrome. MATERIALS AND METHODS: A total of 53 patients with chronic prostatitis treated at our clinic with a minimum followup of 6 months were assessed by the National Institutes of Health-Chronic Prostatitis Symptom Index and by a global assessment score. Treatments included antibiotics, prostatic massage, anti-inflammatory phytotherapy, alpha-blockers and neuromuscular agents. RESULTS: Mean age patient was 45 years and median symptom history was 3.5 years. Based on localizing cultures, and microscopy of urine and prostatic fluid 13% of the cases were category II, 41% were category IIIa and 46% were category IIIb. Mean followup from the last visit was 417 days (range 185 to 1,247). Mean changes +/- SE from the initial to the final score on the National Institutes of Health-Chronic Prostatitis Symptom Index were 10.4 +/- 3.3 to 5.9 +/- 4.4 for pain, 4.2 +/- 2.9 to 2.0 +/- 2.7 for urinary, 8.2 +/- 2.9 to 4.7 +/- 3.4 for quality of life and 22.7 +/- 6.6 to 13.2 +/- 9.5 for total score (p <0.0001). Based on a global subjective assessment 43 of the patients (80%) were better, 8 were the same and 3 were worse. At final assessment 39% of the patients were on no therapy, 22% were on an alpha-blocker, 37% were on quercetin, 13% were on neuromuscular agents and 9% were on antibiotics. CONCLUSIONS: An approach using stepwise therapy with antibiotics, anti-inflammatories and neuromuscular agents can be successful in the majority of patients with long-standing chronic prostatitis. PMID- 12629374 TI - A randomized controlled trial comparing transurethral resection of the prostate, contact laser prostatectomy and electrovaporization in men with benign prostatic hyperplasia: analysis of subjective changes, morbidity and mortality. AB - PURPOSE: We analyze subjective changes, morbidity and mortality in men with lower urinary tract symptoms associated with benign prostatic hyperplasia (BPH) after transurethral resection of the prostate, contact laser prostatectomy and electrovaporization. MATERIALS AND METHODS: A prospective, randomized controlled trial was conducted on men with lower urinary tract symptoms, who met the criteria of the International Scientific Committee on BPH, had a prostate volume between 20 and 65 ml., and had Schafer's obstruction grade 2 or greater. Objective morbidity was recorded for up to 12 months. Subjective morbidity was measured by a questionnaire completed by patients. Subjective changes were quantified using the International Prostate Symptom Score, Symptom Problem Index, Quality of Life question and BPH Impact Index. These indexes and the morbidity questionnaire were measured weekly for the first 6 weeks postoperatively and then at 3, 6 and 12 months. RESULTS: Transurethral prostatic resection was analyzed in 50 men, laser treatment in 45 and electrovaporization in 46. Baseline characteristics, and changes in the symptom scores up to 12 months postoperatively were similar. Perioperative blood loss and perforation were greatest in the resection group, and retention was greatest in the laser group. During the first 6 postoperative weeks there was less pain and less hematuria after resection, and less incontinence after laser prostatectomy. CONCLUSIONS: Subjective changes are similar for transurethral prostatic resection, contact laser and electrovaporization. In the first 6 weeks after treatment there are only slight differences in pain, hematuria and incontinence among the therapies. PMID- 12629375 TI - Effects of sildenafil on cardiopulmonary responses during stress. AB - PURPOSE: To investigate possible effects of sildenafil on the cardiopulmonary responses during sexual intercourse we evaluated cardiopulmonary responses during exercise in a group of impotent patients. MATERIALS AND METHODS: The study sample included patients with erectile dysfunction who underwent a cardiopulmonary exercise test before and after the administration of 100 mg. sildenafil citrate. Cardiopulmonary exercise test parameters at rest, at the anaerobic threshold, at peak exercise and at 1-minute recovery were recorded, including systolic and diastolic blood pressure, the heart rate, O2 consumption, CO2 production, ventilation and the respiratory rate. Furthermore, O2 consumption per kg. body weight, the ventilatory equivalent for O2 consumption (ventilation/O2 consumption) and CO2 production (ventilation/CO2 production), the respiratory quotient, metabolic equivalents metabolic equivalents, oxygen pulse (O2 consumption/heart rate) and the change in O2 consumption/change in heart rate were calculated. RESULTS: In 2 of the 43 patients enrolled in the study myocardial ischemia and high blood pressure were detected at rest in 2, respectively, who were excluded from analysis. In the remaining 41 patients with a mean age +/- SD of 52.3 +/- 8.6 years a statistically significant decrease in systolic and diastolic blood pressure was noted after sildenafil use at all stages tested (p <0.002 to 0.001). The heart rate mildly increased after sildenafil use at rest and at peak exercise (p = 0.018). The O2 pulse decreased at the anaerobic threshold (p = 0.003), peak exercise (p = 0.001) and recovery (p = 0.047). In the 11 patients with a mean age of 40.8 +/- 10.12 years who had psychogenic erectile dysfunction the only 2 parameters affected were an increased heart rate and decreased systolic blood pressure at rest, while O2 consumption/heart rate decreased at the anaerobic threshold. In the 18 patients with a mean age of 61.1 +/- 8.9 years who had organic erectile dysfunction and an unremarkable medical history a decrease was noted in systolic and diastolic blood pressure at rest and at peak exercise, and diastolic blood pressure also at recovery, while the heart rate increased at recovery. In the 12 patients with a mean age of 60.16 +/- 9.12 years who had treated cardiovascular disease systolic and diastolic blood pressure decreased at all states and O2 consumption/heart rate at the anaerobic threshold and at peak exercise, while increased values were noted for the respiratory rate at the anaerobic threshold and ventilation/CO2 production at recovery. CONCLUSIONS: Hemodynamic changes after sildenafil administration should be considered minimal in concert with patient health status. Younger patients without signs of systemic atherosclerosis compensate the vasodilatory effect of sildenafil during exercise, while in older patients with vasculogenic erectile dysfunction moderate changes may be noted regardless of cardiovascular disease in the medical history. PMID- 12629376 TI - Identifying patients with type 2 diabetes with a higher likelihood of erectile dysfunction: the role of the interaction between clinical and psychological factors. AB - PURPOSE: We estimated the prevalence of erectile dysfunction in patients with type 2 diabetes and identified subgroups of patients in which the interaction among clinical, psychological and sociodemographic characteristics determined an increased likelihood of erectile dysfunction. MATERIALS AND METHODS: The presence of erectile dysfunction was based on patient self-reporting. Clinical information was collected by participating physicians. The severity of depressive symptoms was investigated using the Center for Epidemiological Studies Depression scale. To evaluate interactions among the variables investigated and identify distinct, homogeneous subgroups of patients with different odds ratios for erectile dysfunction a tree growing technique was used. RESULTS: In the 1,460 patients studied the prevalence of severe and mild-moderate erectile dysfunction was 34% and 24%, respectively. While severe erectile dysfunction was mainly related to the severity of diabetes, mild-moderate dysfunction was independent of clinical variables and only associated with the severity of depressive symptoms. The tree growing technique led to the identification of 6 classes characterized by a marked difference in the prevalence of severe erectile dysfunction of between 19% and 65%. Patients on diet alone showed the lowest prevalence of erectile dysfunction and were considered the reference category, while patients treated with insulin who had neuropathy represented the subgroup with the highest likelihood of erectile dysfunction (OR = 7.2, 95% CI 3.9 to 13.2). In patients treated with oral agents the odds ratio for erectile dysfunction was 2.7 (95% CI 1.8 to 3.9) for those with severe depressive symptoms and 1.9 (95% CI 1.3 to 2.7) for current/former smokers with low depressive symptoms. Patient age, retinopathy and cardiac-cerebrovascular disease were globally predictive variables associated with an increased likelihood of erectile dysfunction. CONCLUSIONS: Our data illustrate the interplay of clinical and psychological factors in determining the risk of erectile dysfunction in type 2 diabetes and can help identify those for whom much greater attention is needed to detect erectile problems. PMID- 12629377 TI - Serial assessment of efficacy and satisfaction profiles following penile prosthesis surgery. AB - PURPOSE: Penile implants are associated with a high level of patient satisfaction. Studies to date have failed to use instruments that accurately record patient satisfaction data. We used 2 validated instruments to assess the chronology of efficacy and satisfaction profiles in men undergoing penile prosthetic surgery. MATERIALS AND METHODS: Men who elected penile implant surgery were administered the International Index of Erectile Function (IIEF) questionnaire preoperatively, and 3, 6 and 12 months postoperatively as well as the Erectile Dysfunction Inventory of Treatment Satisfaction (EDITS) questionnaire 3, 6 and 12 months postoperatively. Comparison was done of preoperative and postoperative values for the overall IIEF, IIEF erectile function domain, IIEF satisfaction domain and EDITS. All surgeries were primary and in all patients an inflatable implant device was placed. RESULTS: A total of 96 men with a mean age of 56 years were enrolled in the study. All 12-month scores were statistically significantly higher than baseline scores. The 12-month values were statistically higher than 6-month values for the IIEF satisfaction domain and for EDITS. CONCLUSIONS: This study indicates that implant surgery causes a dramatic improvement in erectile function. It also indicates that satisfaction increases in year 1 after implant surgery with significant improvements in the second half of year 1. Current efforts are aimed at following this cohort of patients in an even more long-term fashion as well as evaluating partner satisfaction profiles. These findings should permit clinicians to provide penile prosthesis candidates with optimistic and realistic expectations for implant device function and postoperative satisfaction. PMID- 12629378 TI - Percutaneous vasectomy: a simple modification eliminates the steep learning curve of no-scalpel vasectomy. AB - PURPOSE: We report a simplified method to avoid the most difficult step of no scalpel vasectomy, while maintaining its minimally invasive advantages. MATERIALS AND METHODS: Using the no-scalpel vasectomy instruments in percutaneous fashion we perform vasectomy in the office setting without fixation of the vas to skin using the ring clamp. The sharp no-scalpel hemostat punctures the skin. The vas is then grasped with the ringed instrument instead of piercing the vas and performing the supination maneuver, as described for no-scalpel vasectomy. RESULTS: Percutaneous vasectomy was performed in 573 men by a single surgeon. In the 35 consecutive cases recently reviewed average operative time was 9.3 minutes with an additional 67 seconds added when a resident performed the procedure on 1 side in 15 cases. As determined by the knuckle of vas pulled through a puncture, average incisional length was 8.4 mm. Patients reported complete recovery in an average of 8.9 days. No major complications occurred. A single case of recanalization (0.17%) was successfully corrected by repeat percutaneous vasectomy. CONCLUSIONS: Percutaneous vasectomy is a minimally invasive option for permanent male sterilization that avoids the difficult aspects of no-scalpel vasectomy. PMID- 12629379 TI - Longitudinal differences in disease specific quality of life in men with erectile dysfunction: results from the Exploratory Comprehensive Evaluation of Erectile Dysfunction study. AB - PURPOSE: We assessed the impact of erectile dysfunction therapy on 1-year health related quality of life using a validated erectile dysfunction specific instrument. MATERIALS AND METHODS: Using an observational erectile dysfunction registry clinical, sociodemographic and health related quality of life information was collected at baseline, and 3, 6 and 12 months later. Only men who reported undergoing erectile dysfunction treatment were included in this analysis sub-sample. Patients were classified as treatment responders based on improvements in International Index of Erectile Function scores. Changes in health related quality of life scores from baseline were compared between responders and nonresponders. RESULTS: The cohort consisted of 89 patients, of whom 40 (45%) responded to therapy by International Index of Erectile Function criteria. Responders and nonresponders had comparable baseline disease severity and health related quality of life. At 1 year responders reported significantly better health related quality of life and greater improvement from baseline in sexual experience (mean change -1.64 versus 3.19) and emotional life (mean -3.01 for responders versus 1.75) domains of the Psychological Impact of Erectile Dysfunction scales (p <0.01). This 4.5 to 5 point difference in mean change score (1/2 SD) was considered moderately clinically significant. CONCLUSIONS: Men who respond to erectile dysfunction treatment report significantly better health related quality of life 1 year after initial presentation for erectile dysfunction than nonresponders. This finding should motivate providers to be more proactive in diagnosing and treating men with erectile dysfunction since successful therapy appears to improve health related quality of life. These data suggest that men in whom primary therapy for erectile dysfunction fails should be offered secondary treatment because many men in this study in whom previous therapies failed reported improved health related quality of life when they began effective secondary treatment. PMID- 12629380 TI - Temporal trends in radical prostatectomy complications from 1991 to 1998. AB - PURPOSE: We performed a retrospective, longitudinal, population based study to ascertain whether radical prostatectomy outcomes improved after the diffusion of surgical innovations during the last decade. MATERIALS AND METHODS: Using a 5% national random sample of Medicare beneficiaries we identified 12,079 men who underwent radical prostatectomy from 1991 to 1998. We analyzed relevant Medicare data to establish length of stay and the rate of in hospital complications (cardiac, respiratory, vascular, wound and genitourinary conditions) as well as the rate of anastomotic stricture, incontinence and impotence through 36 months after surgery. We performed multivariate logistic regression to control for age, race and geographic region when assessing the association of surgery year with outcomes of interest. RESULTS: Between 1991 and 1998 the in hospital complication rate decreased from 38% to 30% and mean length of stay decreased from 8.1 to 5.1 days. Each value had significant regional variation throughout the United States. The 3-year incontinence rate decreased from 20% in 1991 to 4% in 1995. However, no meaningful trends were observed in the rate of impotence, anastomotic stricture, or placement of artificial urinary sphincters or penile prostheses. On multivariate analysis, older age (75 years or older, OR 1.68, p <0.01) and nonwhite race (OR 1.35, p <0.01) were associated with more in hospital complications. Nonwhite patients were also more likely to be diagnosed with impotence (OR 1.25, p <0.01) and undergo penile prosthesis placement (OR 1.5, p <0.01). CONCLUSIONS: As urologists reach consensus on the ideal clinical characteristics for radical prostatectomy candidates, surgery in fewer elderly patients and the dissemination of surgical advances have been associated with shorter length of stay, fewer in hospital complications and a lower long-term incontinence rate. However, there is capacity for improvement, as evidenced by the unchanging rate of anastomotic stricture and impotence. PMID- 12629381 TI - Cancer fear and mood disturbance after radical prostatectomy: consequences of biochemical evidence of recurrence. AB - PURPOSE: Biochemical recurrence of prostate cancer often precedes any clinical sign or symptom of disease recurrence by several years. Thus, patients may have laboratory evidence of recurrence and do not know what it portends in terms of the future disease course. Little is known about the emotional consequences of biochemical recurrence. We compared cancer fear and mood disturbance in men with biochemical recurrence of prostate cancer versus those without recurrence. In addition, associations among urinary symptoms, cancer fear and mood disturbance were examined. MATERIALS AND METHODS: A survey including the American Urological Association symptom index, a cancer fear measure and the Profile of Mood States was mailed to patients at a urology clinic at a tertiary care hospital in 1999. Of the sample of 270 patients with prostate cancer who underwent radical prostatectomy 126 (47%) responded to the mailed survey. A total of 45 men with biochemical prostate cancer recurrence were compared to 81 patients without recurrence. RESULTS: Higher urinary tract symptoms were associated with increased cancer fear and mood disturbance (each p <0.05). Biochemical cancer recurrence was not independently associated with increased cancer fear and mood disturbance. However, men with biochemical recurrence and more severe urinary tract symptoms reported the highest levels of cancer fear and mood disturbance (each p <0.01). CONCLUSIONS: Psychological distress was highest in men with biochemical recurrence and elevated clinical symptoms. Urinary symptoms may be an important contributor to psychological distress in patients with prostate cancer who have biochemical recurrence. PMID- 12629382 TI - Determinants of long-term sexual health outcome after radical prostatectomy measured by a validated instrument. AB - PURPOSE: We combined the strengths of previous patient reported studies (that is use of a validated instrument) with the assets of previous single surgeon, physician reported series (that is prospective collection of operative data) by performing a multiple surgeon study to identify demographic and operative determinants of post-prostatectomy sexual health related quality of life outcomes. MATERIALS AND METHODS: Sexual outcome was measured after prostatectomy in 671 prostate cancer cases with a followup of 4 to 52 months treated by any of 7 urologists at a high volume medical center and 112 age matched controls by cross-sectional health related quality of life assessment using the Expanded Prostate Cancer Index Composite validated questionnaire. Multivariable ANCOVA was done to identify the baseline determinants affecting post-prostatectomy sexual health outcomes among 17 clinical and demographic factors. RESULTS: Factors independently associated with better sexual health outcome in patients included younger age (p <0.0001), nerve sparing technique (p <0.0001), time since prostatectomy (p = 0.0001), smaller prostate size (p = 0.003), higher education level (p = 0.02), and higher household income (p = 0.02). Orgasm was achieved at a higher rate than erection. In controls only age (p = 0.0004) and having a partner (p = 0.04) were significantly associated with sexual health. CONCLUSIONS: Larger prostate size adversely affected sexual health outcome after radical prostatectomy independent of all other measured factors. Patient sexual outcome did not differ significantly among surgeons who performed a broad range of such procedures yearly, suggesting that surgical expertise cannot be measured simply by the number of such operations that a surgeon performs. Stratifying patient reported sexual function by the principal determinants of age, nerve sparing and prostate size provides a basis for counseling patients. PMID- 12629383 TI - Is quality of life different for men with erectile dysfunction and prostate cancer compared to men with erectile dysfunction due to other causes? Results from the ExCEED data base. AB - PURPOSE: To our knowledge the relationship between the underlying etiology of erectile dysfunction and its impact on health related quality of life has not been studied. Such a study is important for men with prostate cancer, as the potential negative quality of life impact of erectile dysfunction may affect clinical decision making in newly diagnosed disease. We compare health related quality of life in impotent men with prostate cancer to that of impotent men without prostate cancer using the Exploratory and Comprehensive Evaluation of Erectile Dysfunction (ExCEED, TAP Pharmaceutical Products, Inc., Lake Forest, Illinois) data base, which is a multicenter, observational disease registry of men with erectile dysfunction. MATERIALS AND METHODS: The cohort included 168 men in ExCEED who had baseline health related quality of life measurement. Of these men 47 reported a history of prostate cancer while 121 did not. Appropriate univariate and multivariate analyses were performed comparing health related quality of life outcomes between impotent men with and without prostate cancer. RESULTS: Men with erectile dysfunction and prostate cancer had worse sexual self efficacy, erectile function, intercourse satisfaction and orgasmic function than those with erectile dysfunction without prostate cancer (all p <0.001). However, men with erectile dysfunction and prostate cancer experienced less psychological impact of erectile dysfunction on sexual experience (p = 0.05) and emotional life (p = 0.03) than those with erectile dysfunction without prostate cancer. The findings regarding the psychological impact of erectile dysfunction persisted in multivariate linear regression models. CONCLUSIONS: Men with erectile dysfunction and prostate cancer appear to have better disease specific health related quality of life than those with erectile dysfunction and no history of prostate cancer. This finding has important ramifications for clinicians when counseling patients newly diagnosed with prostate cancer and also when treating patients who present with erectile dysfunction of various etiologies. PMID- 12629384 TI - A prospective study measuring penile length in men treated with radical prostatectomy for prostate cancer. AB - PURPOSE: Some patients report that the penis is smaller after radical retropubic prostatectomy for prostate cancer. We performed a prospective study which measured penile length before and after radical retropubic prostatectomy. MATERIALS AND METHODS: A total of 124 men consented to penile measurements before radical prostatectomy performed by the same surgeon. Repeat measurements were performed at 3-month intervals following surgery. Penile measurements (cm.) consisted of flaccid length, stretched length, depth of prepubic fat pad and circumference. Statistical analysis of penile measurements was performed using the paired samples and independent Student's t test, Pearson's correlation and analysis of variance. RESULTS: Mean patient age was 59.1 (range 42 to 76). No patient had a penile abnormality, for example Peyronie's disease, or history of penile or urethral surgery. The size of the penis was significantly smaller after prostatectomy, and there was a significant difference for flaccid (p <0.001), stretched (p <0.001), prepubic fat pad (p = 0.02) and circumference measurements (p <0.01). Twelve patients (19%) had a 15% or greater decrease in stretched penile length. When dichotomizing groups at the cut point of 15% decrease, the independent t test of prostate size was not significant (t[df 36.7] = -1.83, p = 0.076). Nerve sparing was not significant in relation to change in penile stretched length, (F[df 2, 62] = 0.501, p = 0.609). CONCLUSIONS: Our findings support observations of decreased penile length after radical prostatectomy. Men should be counseled before radical prostatectomy that penile shortening may occur. PMID- 12629385 TI - Editorial: sexual function after radical prostatectomy. PMID- 12629386 TI - A simplified method of implanting a neuromodulator device. AB - PURPOSE: The InterStim neuromodulator device (Medtronic, Inc., Minneapolis, Minnesota) is indicated for the treatment of refractory urge incontinence, urinary frequency and urgency, and nonobstructive urinary retention. We present a modification of the 2-stage approach which is simple to perform, reduces the number of incisions from 2 to 1, decreases operative time and potentially decreases the risk of infection. MATERIALS AND METHODS: Using a single paramedian incision, a quadripolar lead is placed into the appropriate sacral foramen. The lead is connected to the extension wire and the connector is anchored into an ipsilateral subcutaneous tunnel with the aid of an externalized polypropylene suture and plastic button. The extension wire is then externalized through the contralateral buttock using a subcutaneous tunneling device. At stage 2 the entire system can be removed through the original incision or the pulse generator can be implanted through a virgin incision over the connector site. RESULTS: A total of 20 procedures have been performed with an average stage 1 implant time of 72 minutes. Median implant time of 13 pulse generators was 36 minutes. Median explant time of 6 leads was 21 minutes. There have been no infections or adverse events. CONCLUSIONS: This modification is simple, efficient, safe and involves use of tools already familiar to urologists. By reducing the number of incisions, it reduces violation of the skin barrier and consequently risk of infection. Infection is potentially reduced at stage 2 because there is undisturbed, noncontused virgin tissue at the site of generator implant. PMID- 12629387 TI - Erdheim-Chester disease: case report and review of associated urological, radiological and histological features. PMID- 12629388 TI - Tuberculosis of seminal vesicles as a cause of aspermia. PMID- 12629389 TI - Extratesticular inflammatory myofibroblastic tumor. PMID- 12629390 TI - Re: Optimal timing of initial postnatal ultrasonography in newborns with prenatal hydronephrosis. PMID- 12629391 TI - Re: The prevalence of testicular microlithiasis in an asymptomatic population of men 18 to 35 years old. PMID- 12629392 TI - Re: Comparison of hand assisted and standard laparoscopic radical nephroureterectomy for the management of localized transitional cell carcinoma. PMID- 12629393 TI - Re: Bilateral laparoscopic inguinal hernia repair can complicate subsequent radical retropubic prostatectomy. PMID- 12629394 TI - Re: Demographic and clinical characteristics of men with chronic prostatitis: the National Institutes of Health Chronic Prostatitis Cohort study. PMID- 12629395 TI - Prenatally diagnosed ruptured vesico-allantoic cyst presenting as patent urachus at birth. PMID- 12629396 TI - The cost-effectiveness of endoscopic injection of dextranomer/hyaluronic acid copolymer for vesicoureteral reflux. AB - PURPOSE: Vesicoureteral reflux is a risk factor for progressive renal damage associated with urinary tract infection. Mild to moderate reflux is routinely treated with long-term antibiotic prophylaxis to prevent recurrent infections and open surgical reimplantation for breakthrough infections despite antibiotic therapy. Endoscopic subureteral injection of implant material is a therapeutic alternative to long-term prophylaxis and open surgery but its widespread use in the United States has been prevented by the lack of a stable implant material. Dextranomer/hyaluronic acid copolymer has been shown to be a safe, effective and durable implant material and was recently approved in the United States. We estimate the effect on costs and cure rates of introducing endoscopic injection with dextranomer/hyaluronic acid copolymer as a treatment alternative in the United States. MATERIALS AND METHODS: We constructed a model that mimics current clinical practice of vesicoureteral reflux treatment for 6 years, and incorporates spontaneous resolution and surgical intervention rates obtained from 2 long-term followup studies. The treatment algorithm was established using medical data from the literature, and clinical management practices from a Delphi survey of 27 pediatric urologists and nephrologists across the United States. Endoscopic injection was introduced into the model as replacement to surgery or alternative to long-term antibiotic prophylaxis. The effectiveness of dextranomer/hyaluronic acid copolymer was calculated from 140 patients (208 ureters) with grade III reflux treated in a clinical study of 221 children in Sweden. RESULTS: With current practice, the average cost per patient in 6 years was 6,640 US dollars and 23.5% of patients continued to have reflux. Replacing open surgery with endoscopic injection led to similar cure rates (22.2% failures) but costs were reduced to 5,522 US dollars. When injection was performed after 1 year of antibiotic therapy failure rates were reduced to 8.5% but costs increased to 7,644 US dollars. CONCLUSIONS: Our results show that a persistent approach to endoscopic surgery can be expected to result in overall success that equals or exceeds open surgery at a lower cost. This finding is particularly true if open reimplant is reserved for patients with high grade or persistent vesicoureteral reflux. PMID- 12629397 TI - Exploration for testicular remnants: implications of residual seminiferous tubules and crossed testicular ectopia. AB - PURPOSE: Testicular remnants identified during exploration for cryptorchidism contain vascularized fibrous nodules at the termination of the vas deferens, hemosiderin, calcification, a pampiniform plexus or occasionally residual seminiferous tubules that may contain germ cells. An absent testis lacks the features of testicular remnants. To our knowledge testicular remnants have not been described in a crossed ectopic location. We reviewed orchiectomy specimens obtained at exploration for a nonpalpable testis to characterize the features of testicular remnants, including the frequency of seminiferous tubules, germ cells and crossed ectopia, as well as to clarify the diagnostic criteria for testicular remnants. MATERIALS AND METHODS: From 1990 to mid 2000 medical records and histological slides from 101 boys with nonpalpable testes who had undergone inguinal exploration and orchiectomy were reviewed. RESULTS: Of the 71 testicular remnants identified 7 (9.8%) contained residual tubules, of which 4 (5.6%) contained germ cells. In 4 boys the testis was deemed absent but 3 did not undergo laparoscopic exploration. There were 2 ectopic remnants (2.8%) on the contralateral side-the pelvis or in the scrotum. Both crossed remnants demonstrated dissociation of the testis from the vas/epididymis which remained on the correct side associated with a pampiniform plexus. No mullerian remnants were encountered. CONCLUSIONS: Adequate exploration for nonpalpable testis requires laparoscopy with visualization of the contralateral pelvic region because an ectopic remnant may be dissociated from the vas/epididymis and vessels. Identification of a pampiniform plexus, vas and spermatic vessels may not be a reliable indicator of a testicular remnant. Continued removal of testicular remnants is warranted because at least 9.8% contain residual viable tubules. PMID- 12629398 TI - Complications of pediatric urological laparoscopy: mistakes and risks. AB - PURPOSE: We evaluate the results and complications of laparoscopic urological procedures in children. MATERIALS AND METHODS: In a 3-year period 4,350 laparoscopic procedures were performed at 8 Italian centers of pediatric surgery. We analyzed only the data of urological procedures for a total of 701 laparoscopic operations on patients 1 month to 14 years old. The indications for surgery were cryptorchidism in 414 cases, varicoceles in 159, ambiguous genitalia in 37, total nephrectomy in 34, partial nephrectomy in 4, adrenalectomy in 3 and other diagnostic procedures in 50. We adopted a retroperitoneoscopic approach in 72 cases (10.3%) and a laparoscopic approach in 629 (89.7%). Patient records were analyzed to search for any complication that may have occurred during the laparoscopic procedure and assess how they were managed. RESULTS: We recorded 19 complications (2.7%) in our series, of which 6 required conversion to open surgery and 13 did not. There was no mortality. At a maximum followup of 4 years all children were alive and had no problems related to the laparoscopic complications. CONCLUSIONS: Our study shows that pediatric laparoscopic urological surgery has an acceptable rate of complications with no mortality. We believe that routine use of open laparoscopy in pediatric patients is a key factor to help avoid complications. Most complications can be avoided with surgeon and team experience, together with proper compliance with the indications for surgery. PMID- 12629400 TI - Prenatal ultrasonographic detection of bladder rhabdomyosarcoma. PMID- 12629399 TI - Enucleative surgery in an infant with giant cystic nephroma. PMID- 12629401 TI - Juvenile granulosa-theca cell (sex cord-stromal) tumor of the infant testis. PMID- 12629403 TI - Longitudinal and thickness measurement of the normal distal and intravesical ureter in human fetuses. AB - PURPOSE: We define reference data concerning the development of the ureterovesical junction in fetuses and newborns by measuring the diameters of the distal mesenchymal and muscular ureteral walls as well as intravesical ureteral length. MATERIALS AND METHODS: A total of 90 normal fetal and newborn ureters were investigated. Our histological studies were based on "plastinated" sections of whole pelves which allow study of the sectional anatomy of the distal and intravesical ureter. The development of the mesenchymal and smooth muscle growth of the distal and intravesical ureter was examined. The ureteral measurements were correlated with age of gestation. RESULTS: The length of the intravesical ureter and mesenchymal as well as smooth muscle walls increased in a linear mode. Significant correlations (p <0.0001) were found between gestational age and the growth of the mesenchymal as well as smooth muscle walls in the distal intravesical ureter as well as the length of the intravesical ureter. CONCLUSIONS: Significant positive linear relationships exist between gestational week, and distal and intravesical ureteral wall thickness of the mesenchymal and smooth muscle growth to the length of the intravesical ureter in fetuses and newborns. The ratio of the intravesical ureteral length-to-ureteral diameter is obviously lower than assumed previously. Data from this study can be used for a more accurate assessment of cases with abnormal lower urinary tract development. PMID- 12629404 TI - Home uroflowmetry for the evaluation of boys with urinary incontinence. AB - PURPOSE: We recorded uroflowmetry at home in boys with urinary incontinence and correlated the results with videourodynamics. MATERIALS AND METHODS: Thirty-nine boys (mean age 8.4 +/- 2.0 years) with urinary incontinence underwent home uroflowmetry for 1 weekend. Artifactual spikes in 1 or more uroflow curves were present in 16 home uroflowmetry recordings. One patient, in whom none of the uroflow curves was interpretable, was excluded from the study. Of the remaining 38 boys 18 had monosymptomatic nocturnal enuresis, and 20 had nocturnal enuresis and diurnal voiding symptoms. Percentage expected bladder capacity is defined as functional/expected bladder capacity x 100%. Normal and obstructive home uroflowmetry levels are defined as functional bladder capacity at least 50% expected bladder capacity associated with multiple bell-shaped and obstructive uroflow curves, respectively. Small functional bladder capacity is defined as capacity less than 50% expected bladder capacity, regardless of uroflow patterns. Videourodynamics and cystoscopy were performed in 17 patients. RESULTS: Normal home uroflowmetry was noted in 5 patients (13%), obstructive uropathy in 8 (21%) and small functional bladder capacity in 25 (66%). Urodynamically 3 boys with normal home uroflowmetry had normal voiding, and 6 with obstructive home uroflowmetry had bladder outlet obstruction (of whom 1 also had detrusor overactivity). In addition, of 8 boys with small functional bladder capacity 4 had detrusor overactivity, 3 had bladder outlet obstruction and 1 had both findings. CONCLUSIONS: Normal home uroflowmetry predicted normal voiding, and abnormal recordings implied abnormal voiding function in boys with incontinence. Bladder outlet obstruction and detrusor overactivity were frequently disclosed by obstructive home uroflowmetry and small functional bladder capacity. PMID- 12629402 TI - Percutaneous nephroscopic removal of obstructing fungal bezoars and endopyelotomy in an infant. PMID- 12629405 TI - Management of urinary extravasation in 18 boys with posterior urethral valves. AB - PURPOSE: Urinary extravasation with associated posterior urethral valves is uncommon and published articles are restricted to case reports. We propose a management algorithm on the basis of presentation of 18 boys with this condition. MATERIALS AND METHODS: In this retrospective review we analyze the clinical features, diagnostic radiological imaging (voiding cystourethrogram and (99m)technetium mercaptoacetyltriglycine scan) and measured glomerular filtration rate (GFR) for assessment of renal function in boys presenting with posterior urethral valves and urinary extravasation. Indications for surgical intervention and long-term outcome were evaluated. Results are presented as mean +/- SD. RESULTS: Patient age at presentation was 4 days (range 1 to 28). Salient features at presentation were abdominal distention in 55.5% of cases, flank mass in 55.5%, respiratory distress in 16.6% and septicemia in 22.2%. Patients were categorized into group 1 with ascites (6 boys) and group 2 without ascites (12). Followup was 70.8 +/- 5.6 months. In group 1 the 3 boys with ascites alone had a followup GFR of 29 versus 74 ml. per minute per 1.73 m.2 in 3 with urinoma plus ascites (p = 0.001) at 64 +/- 4.9 months. In group 2 the GFR in 3 boys with bilateral urinoma was 104.7 +/- 30.4 versus 54.3 +/- 29.4 ml. per minute per 1.73 m.2 in the other 9 with unilateral urinoma (p = 0.001) at 36 +/- 4.2 and 81.3 +/- 4.7 months, respectively. The ipsilateral kidney showed reduced function in 6 of 12 boys with unilateral urinoma from both groups. CONCLUSIONS: Renal function on the side of the urinoma is impaired, and the majority of patients have impaired global renal function. A management algorithm is proposed. PMID- 12629406 TI - In vitro fertilization and the cloacal-bladder exstrophy-epispadias complex: is there an association? AB - PURPOSE: The cloacal-bladder exstrophy-epispadias complex represents a collection of congenital malformations caused by failed mesenchymal development during the first trimester. We sought to determine whether the exstrophy-epispadias complex is overrepresented among children conceived by in vitro fertilization (IVF). MATERIALS AND METHODS: The clinic charts of all patients born between 1998 and 2001 who presented to our institution with the exstrophy-epispadias complex were reviewed. Four patients conceived by IVF and born with exstrophy-epispadias were identified and telephone interviews were conducted with the parents. Statistical analysis using clinical data and available United States population data was performed. RESULTS: Of 78 clinic patients with exstrophy-epispadias born during a 4-year period 4 were conceived using IVF. An estimated 12% to 14% of children born in the United States with the exstrophy-epispadias complex are evaluated annually at our institution. During the years 1997 to 2000, 112,137 children in the United States were conceived using IVF. According to published incidence data, approximately 5 children with exstrophy-epispadias would be expected among this entire population. Expected numbers of children in the United States conceived by IVF and born with exstrophy-epispadias during a 4-year period were calculated based on the observed number in our clinic population. A resulting 7.3 fold relative increase in incidence was determined (p = 0.0021). CONCLUSIONS: The exstrophy-epispadias complex appears to occur more frequently in children conceived by in vitro fertilization. PMID- 12629407 TI - Doxazosin and terazosin suppress prostate growth by inducing apoptosis: clinical significance. AB - PURPOSE: Doxazosin and terazosin are known to relax prostate smooth muscle through blockade of alpha 1-adrenergic innervation to the prostate. This action alone however does not fully account for the long-term clinical responses exerted by these drugs in the treatment of patients with benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: Experimental and clinical studies were done to establish the induction of prostate cell apoptosis by alpha 1-adrenoceptor antagonists as a molecular mechanism contributing to their long-term efficacy in the management of lower urinary tract symptoms associated with BPH and to potential suppression of prostate cancer growth. RESULTS: The data indicate that both doxazosin and terazosin induce apoptosis in prostate cancer cells in vitro and in vivo. The apoptotic effect of doxazosin and terazosin is mediated by a mechanism independent of the alpha1-adrenoceptor blockade, potentially under the direction of the quinazoline nucleus, since the nonquinazoline alpha 1 adrenoceptor antagonist tamsulosin does not elicit an apoptotic response. Recent experimental evidence points to deregulation of signal transduction pathways involving transforming growth factor-beta and disruption of cell attachment to the extracellular matrix (anoikis) as potential mechanisms underlying this apoptotic action of quinazoline based alpha 1-adrenoceptor antagonists against prostate cells. CONCLUSIONS: The correlation of induced prostate smooth muscle cell apoptosis with improvement of urinary symptoms in patients with BPH treated with doxazosin and terazosin, identifies apoptosis as an additional molecular mechanism for the long term therapeutic impact of these drugs in BPH. Moreover, the apoptotic effect elicited by quinazolines may have high clinical significance in the prevention and treatment of prostate cancer. PMID- 12629408 TI - Shock wave lithotripsy causes ipsilateral renal injury remote from the focal point: the role of regional vasoconstriction. AB - PURPOSE: Shock wave lithotripsy induced renal damage can occur as a result of multiple mechanisms, including small vessel injury and free radical production. Previous studies have demonstrated that shock wave lithotripsy exerts a regional change in renal hemodynamics, resulting in a global reduction in the glomerular filtration rate and renal plasma blood flow. We determined if biochemical evidence of cellular damage could be identified in ipsilateral locations remote from the shock wave site or in the contralateral kidney, suggesting regional or systemic alterations in renal function. MATERIALS AND METHODS: Ten juvenile female swine underwent open insertion of microdialysis probes into the renal parenchyma at the right upper and lower poles, and left lower pole. The animals were divided evenly into a sham and a treatment group. Dialysate samples were collected from all 3 sites from the sham group at 10-minute intervals for 100 minutes and quantitatively assessed for conjugated dienes, a measure of lipid peroxidation and free-radical activity, signifying renal cellular damage. The animals in the treatment group underwent shock wave lithotripsy focused on the right lower pole. Dialysate samples were collected from all 3 sites at baseline and at 1,000 shock intervals for a total of 10,000 shocks and analyzed for conjugated dienes. The results from the sham and treatment groups as well as from the different locations within each group were compared using Student's t test. RESULTS: The mean conjugated diene ratio for the sham group was 3.59, 3.42 and 2.7 microM. for the right upper and lower poles, and left kidney (p >0.05). A dose related increase in conjugated diene ratio levels from the right lower pole (lithotripsy site) and to a lesser degree from the ipsilateral right upper pole were observed, which were significantly different from sham group measurements or the contralateral kidney of the treatment group (p <0.05). The elevation in conjugated diene levels at the lithotripsy site was also noted to be significantly greater than that of the upper pole of the ipsilateral kidney (p <0.05). CONCLUSIONS: The increase in free radical activity at a site remote from the treated region suggests detrimental global effects from shock wave lithotripsy therapy. These observations could be the result of vasoconstriction throughout the treated kidney with resultant ischemia-reperfusion injury. Although these global renal effects may subject patients with baseline renal dysfunction to irreversible renal damage, the clinical significance of our findings is unclear and warrants further investigation. PMID- 12629409 TI - Matrix metalloproteinase activity in urine of patients with renal cell carcinoma leads to degradation of extracellular matrix proteins: possible use as a screening assay. AB - PURPOSE: Localized renal cell carcinoma is usually curable by nephrectomy. However, a large fraction of patients already present with metastatic disease, which results in a poor outcome. Currently no clinically relevant screening assay is available to detect early stage renal cell carcinoma. We investigated whether urinary extracellular matrix (ECM) proteins and/or matrix metalloproteinase (MMP) activity may be valuable as a noninvasive indicator of early stage renal cell carcinoma. MATERIALS AND METHODS: Urine specimens from preoperative patients with renal cell carcinoma and healthy controls were collected. The urinary excretion of the ECM proteins collagen IV, laminin and fibronectin was investigated by immunoblotting. MMP activity was assessed by gelatin zymography and by a fluorescence based microtiter plate activity assay. RESULTS: The full-length forms of all 3 ECM proteins investigated were significantly decreased or absent in renal cell carcinoma urine. Based on criteria established in this study this finding would lead to the correct detection of 95% of patients with renal cell carcinoma (21 of 22) with a false-positive rate of 4.5% (1 of 22 controls). All 11 nonmetastatic cases of the lowest clinical stage (T1N0M0) were correctly identified. The absence of urinary ECM proteins was due to significantly increased urinary MMP activity. CONCLUSIONS: Analysis of decreased urinary ECM proteins and analysis of increased MMP activity may have value for the development of a sensitive, high throughput molecular screening assay to detect early stage renal cell carcinoma. PMID- 12629410 TI - Targeting urothelium: ex vivo assay standardization and selection of internalizing ligands. AB - PURPOSE: With the goal of targeting the human bladder using phage display technology we designed and tested a tissue binding assay on intact urothelium ex vivo. This approach may form the molecular basis for clinical development of peptide or peptidomimetic guided intravesical compounds. MATERIALS AND METHODS: We screened 2 phage display random peptide libraries on human urothelium. Select peptides were tested for their binding ability to human urothelium, 2 human transitional cell carcinoma cell lines and a nontransitional cell carcinoma cell line. Next we standardized an ex vivo binding assay, validated binding of selected phage to whole urothelium, and evaluated whether receptor mediated internalization into urothelium derived cells occurred. Finally we tested if the presence of the glycosaminoglycan layer had any effect on the binding of the urothelium targeted phage. RESULTS: Phage selected and recovered in the screening were isolated and sequenced. Displayed peptide sequences were searched against online protein databases. Five classes of peptide motifs were characterized based on their ability to bind to normal urothelium but not to control cell lines. Remarkable consistency and reproducibility were observed in the ex vivo binding assays. Two classes of peptide motifs sharing the sequence Ile/Leu-Ser-Gly-Leu bound to normal urothelium and to 2 transitional cell carcinoma cells but not to nontransitional cell carcinoma cells in a glycosaminoglycan independent manner and mediated internalization into cells of urothelial origin. CONCLUSIONS: We introduce a strategy for screening combinatorial peptide libraries on bladder mucosa, a standard model for ex vivo intact urothelium binding assays and a panel of urothelium binding peptides that may be suitable for translation into targeted intravesical therapy applications. PMID- 12629411 TI - Hypermethylation of an E-cadherin (CDH1) promoter region in high grade transitional cell carcinoma of the bladder comprising carcinoma in situ. AB - PURPOSE: We elucidated the role of methylation in the promoter region of the 1 gene in bladder carcinogenesis, particularly in those comprising carcinoma in situ. MATERIALS AND METHODS: A total of 49 cases of transitional cell carcinoma of the bladder obtained from transurethral resection were examined. Methylation status of the 1 promoter region was analyzed by methylation specific polymerase chain reaction from chemically modified DNA after Na-bisulfite treatment. Loss of heterozygosity on 16q was examined by blunt end single strand DNA conformation polymorphism using 4 tetranucleotide repeat microsatellite markers assigned on 16q13 to 22.1. E-cadherin expression was evaluated by immunostaining on formalin fixed, paraffin embedded tissue sections using anti E-cadherin murine monoclonal antibody, HECD1 and standard avidin-biotin immunoperoxidase complex technique. RESULTS: Analysis of the 49 bladder transitional cell carcinoma samples showed 1 promoter methylation in 23 (47%). Methylation of the 1 gene did not correlate with tumor stage (p = 0.2097) but with high grade transitional cell carcinoma (p = 0.0416). 1 promoter methylation was observed at a significantly higher frequency in the carcinoma in situ positive group than in the carcinoma in situ negative group (16 of 18 cases or 89% versus 7 of 31 or 23%, p <0.0001) and it strongly correlated with abnormal E-cadherin expression (p <0.0001). We found 16q loss of heterozygosity in 16 of 47 cases (34%), which correlated with higher histological grade (p = 0.0069) but not with the presence of the carcinoma in situ component (p = 0.1235). CONCLUSIONS: This study showed that 1 gene promoter methylation is strongly associated with bladder transitional cell carcinoma comprising carcinoma in situ. PMID- 12629412 TI - The roles of supernumerical X chromosomes and XIST expression in testicular germ cell tumors. AB - PURPOSE: An overabundance of X chromosomes in testicular germ cell tumors and the identification of the candidate testicular germ cell tumor susceptibility gene TGCT1 on Xq27 highlight the potential involvement of X chromosomes in testicular germ cell tumor pathogenesis. The current study was designed to shed light on the question whether the multiple X chromosomes in testicular germ cell tumor are active or inactive through a complex mechanism of X chromosomal gain and XIST expression. MATERIALS AND METHODS: We analyzed 4 testicular germ cell tumor derived cell lines and 20 primary testicular germ cell tumor tissues. The number of X chromosomes was determined by fluorescence in situ hybridization using the X chromosome specific probe. The expression patterns of XIST and the 3 X-linked genes androgen receptor (AR), fragile X mental retardation (FMR1 ) and Glypican 3 (GPC3 ) were studied by reverse transcriptase-polymerase chain reaction. Bisulfite genomic sequencing was used to analyze the methylation patterns of the AR, FMR1 and GPC3 genes. The relative expression levels of the 2 X-linked proto oncogenes ARAF1 and ELK1 were assayed by quantitative reverse transcriptase polymerase chain reaction. RESULTS: XIST expression was common in seminomatous testicular germ cell tumors (2 of 2 or 100% of seminoma derived cell lines and 10 of 12 or 83% of seminomatous testicular germ cell tumor tissues) but not in nonseminomatous testicular germ cell tumors (0 of 2 or 0% nonseminoma derived cell lines and 2 of 8 or 25% of nonseminomatous testicular germ cell tumor tissues). However, X chromosomal gain was consistently observed in the 2 types of tumors. XIST expression in testicular germ cell tumors and normal testicular parenchyma was not associated with methylation of the AR, FMR1 or GPC3 genes. After determining the expression patterns of AR, FMR1 and GPC3 in testicular germ cell tumor samples we concluded that multiple X chromosomes in testicular germ cell tumors were predominantly hypomethylated and active regardless of XIST expression. The biological significance of excess active X chromosomes in testicular germ cell tumors was suggested by enhanced expression of the 2 X linked oncogenes ARAF1 and ELK1 in the testicular germ cell tumor derived cell lines. CONCLUSIONS: The current data may suggest the potential oncogenic implications of X chromosomal gain in testicular germ cell tumors. PMID- 12629413 TI - Regulation of Bcl-2 expression by dihydrotestosterone in hormone sensitive LNCaP FGC prostate cancer cells. AB - PURPOSE: Up-regulation of the anti-apoptotic bcl-2 proto-oncogene is associated with androgen independent prostate cancer progression. This observation suggests that the expression of bcl-2 may be negatively regulated by androgens in prostate cancer cells. MATERIALS AND METHODS: The expression of the proto-oncogene bcl-2 was assessed in the hormone sensitive prostate cancer cell line LNCaP-FGC in the presence and absence of a physiological concentration of 1 nM. dihydrotestosterone (DHT). Sequence analysis of the bcl-2 promoter regions demonstrated the presence of 2 potential androgen response elements. Transient transfections of luciferase reporter constructs containing these potential androgen response elements into LNCaP-FGC cells in the presence and absence of DHT were performed. Steady-state transcripts of bcl-2 were assessed using RNase protection assays. RESULTS: Cells cultured in charcoal stripped serum in the presence of DHT resulted in down-regulation of bcl-2 protein. Down-regulation of bcl-2 protein and mRNA by DHT was inhibited by coincubation with the antiandrogen bicalutamide, an agent that competitively inhibits binding of DHT to androgen receptor. Luciferase reporter constructs containing candidate androgen response elements were transrepressed in the presence of DHT. Bcl-2 mRNA was also down regulated by DHT and this down-regulation could not be abolished by cycloheximide. CONCLUSIONS: Together these results suggest that the suppression of bcl-2 expression by DHT in hormone sensitive LNCaP-FGC prostate cancer cells occurs directly. In addition, these results provide a possible mechanistic basis for the up-regulation (derepression) of bcl-2 observed in hormone independent prostate cancers. PMID- 12629414 TI - Editorial: hormone therapies for prostate cancer--acute disease control, chronic disease progression. PMID- 12629415 TI - Novel nuclear factor kappa B activation inhibitor prevents inflammatory injury in unilateral ureteral obstruction. AB - PURPOSE: We determined whether the novel nuclear factor kappa B activation inhibitor dehydroxymethylepoxyquinomicin (DHMEQ), which is derived from epoxyquinomicin C, affects renal inflammatory responses in unilateral ureteral obstruction. MATERIALS AND METHODS: DHMEQ (8 mg./kg.) was administered to rats 1 day after unilateral ureteral obstruction and every day thereafter. Kidneys were harvested at day 7 after unilateral ureteral obstruction. Tissue nuclear factor kappa B activity and transforming growth factor-beta were determined by electrophoretic mobility shift assay and bioassay using mink lung epithelial cells, respectively. Renal tubular proliferation and apoptosis were detected by immunostaining proliferating cellular nuclear antigen and the TUNEL (Intergen, Purchase, New York) assay, respectively. Leukocyte infiltration was detected by immunostaining for CD45. Fibrosis was assessed by measuring tissue hydroxyproline content. RESULTS: Unilateral ureteral obstruction for 7 days significantly activated nuclear factor kappa B, induced tubular apoptosis, proliferation and interstitial fibrosis in the obstructed kidney of the control group compared with their unobstructed counterparts (30.3 +/- 4.5 nuclei per high power field versus 1.7 +/- 0.4, 25.7 +/- 3.3 nuclei per high power field versus 3.2 +/- 0.4 and 6.2 +/- 0.3 micromol. hydroxyproline per gm. tissue versus 3.4 +/- 0.1, respectively). Conversely daily administration of DHMEQ (8 mg./kg.) significantly inhibited nuclear factor kappa B activation and decreased mean tubular apoptosis (9.5 +/- 2.1 nuclei per high power field), proliferation (10.2 +/- 2.4 nuclei per high power field) and interstitial fibrosis (4.9 +/- 0.4 micromol. hydroxyproline per gm. tissue) in the obstructed kidney. CONCLUSIONS: Specific inhibition of nuclear factor kappa B can prevent inflammatory renal responses, suggesting that targeting nuclear factor kappa B activation may be feasible for preventing inflammatory kidney diseases. PMID- 12629416 TI - The ameliorating effect of melatonin on protamine sulfate induced bladder injury and its relationship to interstitial cystitis. AB - PURPOSE: The pineal hormone melatonin was recently shown to have free radical scavenging ability and it reduces lipid peroxidation. In this morphological study we investigated the effects of melatonin on protamine sulfate (Sigma Chemical Co., St. Louis, Missouri) induced bladder injury. MATERIALS AND METHODS: Albino Wistar female rats were catheterized and intravesically infused with phosphate buffered solution (control group) or protamine sulfate (bladder injury group) dissolved in phosphate buffered solution. In the protamine sulfate plus melatonin group after protamine sulfate instillation melatonin was injected intraperitoneally. Bladder morphology was investigated by light and electron microscopy. Tissue samples were also obtained to determine bladder malondialdehyde levels. RESULTS: In the bladder injury group ulcerated areas, an irregular glycosaminoglycan layer, increased number of mast cells, vacuole formation, dilated perinuclear cistern, formation of pleomorphic and uniform microvilli, and dilated urothelial intercellular spaces were observed. In the bladder injury plus melatonin group a relatively normal urothelial topography, glycosaminoglycan layer and decreased number of mucosal mast cells, some dilatation between intercellular areas, less uniform microvilli and in most areas regular tight junctions were observed. CONCLUSIONS: Increased malondialdehyde levels as a result of protamine sulfate induction lead us to propose that free radicals may have a critical role in this injury. The significant decrease in malondialdehyde levels in the protamine sulfate plus melatonin group was in accordance with morphological findings. Thus, melatonin appears to exert a urothelial protective activity in a bladder injury model. PMID- 12629417 TI - Inducible nitric oxide synthase promotes pathophysiological consequences of experimental bladder outlet obstruction. AB - PURPOSE: Bladder outlet obstruction leads to histological and functional changes in the bladder over time. We investigated the role of inducible nitric oxide synthase (iNOS) in the progression of pathological changes of the bladder secondary to outlet obstruction in a rat and a mouse model. MATERIALS AND METHODS: To assess expression of iNOS in the bladder, polymerase chain reaction amplification of mRNA was done. Rats were subjected to sham operation or partial bladder outlet obstruction. They were given the iNOS inhibitor aminoguanidine in drinking water or unmodified water. After 2 weeks, awake cystometric evaluation was performed, the bladders were harvested and the degree of fibrosis was assessed. In another series of experiments mice deficient in the iNOS gene (iNOS /-) were compared to WT mice for cystometric as well as histological changes in the bladder following partial bladder outlet obstruction or sham operation. RESULTS: Partial bladder outlet obstruction induced the expression of iNOS mRNA in the mouse bladder. iNOS -/- mice showed a significantly smaller increase in bladder volume at 3 weeks compared with WT. Pharmacological inhibition of iNOS activity significantly attenuated the increase in bladder size and the number of spontaneous bladder contractions in obstructed rats at 2 weeks. Furthermore, genetic and pharmacological decreases in iNOS led to significantly less fibrosis of the bladder after partial bladder outlet obstruction in mice and rats, respectively. CONCLUSIONS: Pharmacological or genetic decreases in iNOS resulted in amelioration of functional and fibrotic changes in the bladder after partial bladder outlet obstruction, suggesting that NO contributes to the pathophysiology of bladder outlet obstruction. PMID- 12629418 TI - Morphology, phenotype and ultrastructure of fibroblastic cells from normal and neuropathic human detrusor: absence of myofibroblast characteristics. AB - PURPOSE: Fibroblasts are functionally diverse and fibroblastic cells with smooth muscle-like characteristics (myofibroblasts) regulate smooth muscle activity in certain tissues. The presence of myofibroblasts has been reported in the bladder with important implications for normal function and detrusor overactivity. We assessed fibroblastic cell characteristics to discern features suggesting a myofibroblast phenotype in normal or neuropathic human detrusor. MATERIALS AND METHODS: A total of 25 control samples were obtained from cadaveric organ donors or patients with a mean age of 42.3 years investigated for hematuria and compared with samples from 18 patients with a mean age of 37.4 years with neurogenic detrusor overactivity. Morphology, phenotypic expression of various markers and the ultrastructure of each fibroblastic cell visible in multiple sections from each specimen was evaluated by 2 independent assessors. RESULTS: Fibroblastic cells were observed throughout the smooth muscle and connective tissue. They were located peripherally on muscle fascicles and had a polar stellate appearance with processes ramifying in interfascicular planes and muscle. They possessed vimentin like immunoreactivity and weak c-kit-like immunoreactivity but not desmin or alpha-smooth muscle actin-like immunoreactivity. Ultrastructurally they showed dilated rough endoplasmic reticulum with a moderately electron dense amorphous content and prominent golgi complexes. Nuclei had clumped peripheral heterochromatin. There were extensive flattened processes that lacked basal laminae. There was no specific contact with nerve fibers or smooth muscle. Neuropathic bladder samples did not differ overtly from those of controls. CONCLUSIONS: The detrusor possesses an extensive network of fibroblastic cells and processes. No evidence of myofibroblast differentiation was discerned in normal or neuropathic detrusor, although a minority subpopulation or regional variability in cellular phenotype could not be excluded. PMID- 12629419 TI - The effect of vascular endothelial growth factor and adeno-associated virus mediated brain derived neurotrophic factor on neurogenic and vasculogenic erectile dysfunction induced by hyperlipidemia. AB - PURPOSE: We examined neurogenic and vasculogenic erectile dysfunction associated with hypercholesterolemia and evaluated vascular endothelial growth factor (VEGF) and adeno-associated virus (AAV) mediated, brain derived neurotrophic factor (BDNF) for potential treatment. MATERIALS AND METHODS: A total of 21, 2-month-old male rats were fed a 2% cholesterol diet and another seven were fed a normal diet. Two months later serum cholesterol levels were measured and test agents were given intracavernously. Those on normal diet (controls) received phosphate buffered saline (PBS). Those on cholesterol diet were randomly divided into 3 groups receiving PBS, VEGF (4 microg.) or AAV-BDNF (10 viral particles). Four months later erectile function was evaluated and cavernous tissues were collected for erectile dysfunction and immunohistochemical staining. RESULTS: Serum cholesterol levels were higher in rats fed the high fat diet than in controls. Intracavernous pressure was lower in cholesterol plus PBS treated rats than in rats of the other 3 groups. All hypercholesterolemic rats had less nerve content, fewer endothelial cells and higher smooth muscle content than rats with normal cholesterol levels. In cholesterol plus PBS treated rats electron microscopy showed hypermyelination and severe atrophy of axons, a remarkable decrease in the number and size of nonmyelinated axons, disarray of the smooth muscle cells with scant myofilaments and foamy cytoplasm, and denuded endothelial lining of the sinusoids covered by numerous platelets. VEGF and AAV-BDNF appeared to alleviate partially these changes. CONCLUSIONS: A high fat diet caused erectile dysfunction with accompanying neurological and vascular changes. VEGF and AAV-BDNF seemed to alleviate these problems. PMID- 12629420 TI - Exposure to genistein during gestation and lactation demasculinizes the reproductive system in rats. AB - PURPOSE: Exposure to the phytoestrogen genistein (Indofine Chemical Co., Somerville, New Jersey) can disrupt normal male sexual differentiation. To determine if perinatal (that is gestation and lactation) genistein exposure at doses common in human diets alters masculinization we examined the development of the external genitalia, testes, wolffian ducts and sexual behavior in male rats exposed to genistein supplemented diets during early development. MATERIALS AND METHODS: Female rats were fed a phytoestrogen-free diet supplemented with no genistein (free), a low genistein dose (low) or a high genistein dose (high) throughout gestation and lactation. Anogenital distance of male offspring was measured weekly from postnatal days 2 to 21. At puberty (postnatal day 40 to 45) preputial separation, and testis length and width of male offspring were measured. At age 70 days reproductive organ masses, plasma testosterone concentration, sperm counts and sexual behavior were assessed in male offspring. RESULTS Exposure to genistein resulted in temporary, prepubertal urogenital abnormalities at postnatal days 21 and 40. Males exposed to genistein had smaller anogenital distance and testis size, and delayed preputial separation. Perinatal exposure to genistein also caused long-term dysfunction in reproductive behavior, in which adult males exposed to genistein were less likely to mount, intromit and ejaculate during mating tests. Males exposed to genistein also had lower testosterone concentrations in adulthood. CONCLUSIONS: Perinatal genistein exposure results in transient and lasting alterations in masculinization of the reproductive system. These results extend our knowledge of the effects of early genistein exposure on male development and may have implications for human health in terms of potential relationships of endocrine disrupters and urogenital abnormalities thought to be increasing in incidence in boys and men. PMID- 12629421 TI - History of developmental-behavioral pediatrics. PMID- 12629422 TI - Precompetition manipulation: placebo or performance enhancer? PMID- 12629423 TI - Prevention of anterior cruciate ligament injuries in female team handball players: a prospective intervention study over three seasons. AB - OBJECTIVE: To assess the effect of a neuromuscular training program on the incidence of anterior cruciate ligament injuries in female team handball players. DESIGN: Prospective intervention study. SETTING: Female team handball: Division I III in Norway. PARTICIPANTS: Players from the three top divisions: control season (1998-1999), 60 teams (942 players); first intervention season (1999-2000), 58 teams (855 players); second intervention season (2000-2001), 52 teams (850 players). INTERVENTION: A five-phase program (duration, 15 min) with three different balance exercises focusing on neuromuscular control and planting/landing skills was developed and introduced to the players in the autumn of 1999 and revised before the start of the season in 2000. The teams were instructed in the program and supplied with an instructional video, poster, six balance mats, and six wobble boards. Additionally, a physical therapist was attached to each team to follow up with the intervention program during the second intervention period. MAIN OUTCOME MEASURES: The number of anterior cruciate ligament injuries during the three seasons and compliance with the program. RESULTS: There were 29 anterior cruciate ligament injuries during the control season, 23 injuries during the first intervention season (OR, 0.87; CI, 0.50-1.52; p = 0.62), and 17 injuries during the second intervention season (OR, 0.64; CI, 0.35-1.18; p = 0.15). In the elite division, there were 13 injuries during the control season, six injuries during the first intervention season (OR, 0.51; CI, 0.19-1.35; p = 0.17), and five injuries in the second intervention season (OR, 0.37; CI, 0.13-1.05; p = 0.06). For the entire cohort, there was no difference in injury rates during the second intervention season between those who complied and those who did not comply (OR, 0.52; CI, 0.15-1.82; p = 0.31). In the elite division, the risk of injury was reduced among those who completed the anterior cruciate ligament injury prevention program (OR, 0.06; CI, 0.01-0.54; p = 0.01) compared with those who did not. CONCLUSIONS: This study shows that it is possible to prevent anterior cruciate ligament injuries with specific neuromuscular training. PMID- 12629425 TI - The development and evaluation of a disease-specific quality-of-life questionnaire for disorders of the rotator cuff: The Western Ontario Rotator Cuff Index. AB - OBJECTIVE: The purpose of this study was to develop a valid and reliable disease specific quality-of-life measurement tool for patients with rotator cuff disease. DESIGN: Health-related quality-of-life measurement tool development. METHODS: Methodology for the development and evaluation of the tool included the following: 1) identification of a specific patient population, 2) generation of potential items, 3) item reduction, 4) pretesting the prototype instrument, 5) determination of reliability, and 6) validation. RESULTS: The final instrument, the Western Ontario Rotator Cuff Index, double dagger has 21 items representing five domains, each with a Visual Analog Scale-type response option. Construct validation demonstrated that this instrument correlated predictably with other measurement tools (Disabilities of the Arm, Shoulder, and Hand outcome measure; American Shoulder and Elbow Surgeons Standardized Shoulder Assessment Form; University of California Los Angeles Shoulder Rating Scale; Constant Score; Rowe; Sickness Impact Profile; Short Form 36; and range of motion; 21 of 21 correlations within 0.19). Reliability was very high at 2 weeks, with an intraclass correlation coefficient of 0.96 and was more responsive (sensitive to change) than the other five shoulder measurement tools, global health instruments, and range of motion. CONCLUSIONS: This measurement tool can be used as the primary outcome in clinical trials evaluating treatments in this patient population, although its features are equally attractive for monitoring patients' progress in clinical practice. PMID- 12629424 TI - Cross-sectional outcome analysis of athletes with chronic patellar tendinopathy treated surgically and by extracorporeal shock wave therapy. AB - OBJECTIVE: To compare the outcome of chronic patellar tendinopathy treated surgically and by extracorporeal shock wave therapy. DESIGN: Cross-sectional outcome analysis. SETTING: University Hospital Pellenberg, Leuven, Belgium. PATIENTS AND INTERVENTIONS: Of a total of 27 patients (28 knees) with chronic patellar tendinopathy, 13 were treated surgically, and 14 (15 knees) received extracorporeal shock wave therapy. MAIN OUTCOME MEASUREMENTS: Functional outcome questionnaire: Victorian Institute of Sport Assessment score, Visual Analog Scale, and Roles and Maudsley classification. RESULTS: At an average of 6 months of follow-up, the mean Victorian Institute of Sport Assessment and Visual Analog Scale scores for the extracorporeal shock wave therapy group were 78.8 +/- 28.7 and 9 +/- 2, respectively, which improved to 83.9 +/- 28.6 and 9 +/- 2, respectively, at 22.1 months. The surgery group scored 70.7 +/- 22.2 points on the Victorian Institute of Sport Assessment at an average of 26.3 months of follow-up (p = 0.41 and p = 0.18) and 8 +/- 3 on the Visual Analog Scale (p = 0.14). In the extracorporeal shock wave therapy group, 5 (33%) patients rated their pain status at 22.1 months as excellent, 5 (33%) as good, 2 (13%) as fair, and 2 (13%) as poor. In the surgery group, 4 (33%) evaluated the result as excellent, 3 (25%) as good, 3 (25%) as fair, and 2 as poor (17%). CONCLUSIONS: Extracorporeal shock wave therapy shows a comparable functional outcome to surgery in a cross-sectional analysis of patients with chronic proximal patellar tendinopathy resistant to conservative treatment. PMID- 12629426 TI - Comparison of reporting systems to determine concussion incidence in NCAA Division I collegiate football. AB - OBJECTIVE: To determine the incidence of concussion during the 2001 Division I-A college football season through utilization of the Internet. DESIGN: Prospective Internet survey. SETTING: Internet Web site. PARTICIPANTS: Head athletic trainers from Division I-A collegiate football programs. MAIN OUTCOME MEASURES: During the 2001-2002 football season, head athletic trainers from 87 Division I-A football programs agreed to use an Internet Web site to submit weekly data on the number of athlete exposures and concussions. RESULTS: A total of 373 concussions were reported over the course of a season in both practices and games combined. Of these concussions, 256 (68.6%) occurred during games, and 117 (31.4%) occurred during practice. The injury rate per 1,000 athletic exposures was 5.56 in games and 0.25 in practices. There were 230 grade 1 (61.7%), 134 grade 2 (35.9%), and 9 grade 3 (2.4%) concussions reported. There was a significant (p < 0.01) difference found between the total concussion injury rate and game concussion injury rate when comparing the Internet and the NCAA injury data collection methods. CONCLUSIONS: The results of this study suggest an underreporting of concussions using the NCAA Injury Surveillance System and demonstrate that the Internet is a simple and effective tool for data collection. PMID- 12629427 TI - Lactic response in sickle cell trait carriers in comparison with subjects with normal hemoglobin. AB - OBJECTIVE: The objective of this study was to assess lactic response (mmol/L) during incremental exercise in a specific population of men and women with sickle cell trait. SUBJECTS: Athletic men and women from the French West Indies showing no anemia and/or alpha thalassemia. METHODS: Anemia was ruled out by hemoglobin level, and alpha thalassemia was ruled out by hemoglobin S percentage with concomitant hemoglobin level and mean corpuscular volume. Comparison was made with control subjects with normal hemoglobin, matched for physical fitness, anthropometric data, and sex. All subjects performed an incremental exercise test using a cycle ergometer. Peak oxygen uptake (mL/min) was predicted from an equation at peak exercise. Heart rate (beats/min) was noted continuously, and blood samples were collected for lactic response determination every minute during the incremental exercise test and at 1, 3, and 5 minutes of recovery. RESULTS: Comparison of peak oxygen uptake and heart rate values showed no significant differences in subjects with sickle cell trait versus control subjects during the incremental exercise test. In contrast, analysis of variance revealed significantly lower lactic response time courses during the incremental exercise test and at peak exercise (p < 0.05) in the sickle cell trait group, whereas the heart rate time course was lower during recovery (p < 0.05). CONCLUSION: We conclude that the lower lactic response exhibited by subjects with sickle cell trait during incremental exercise was the result of an adaptive process in red blood cells in relation to either lower lactate production or greater lactate uptake capability than in individuals with normal hemoglobin. PMID- 12629428 TI - Practical nonoperative management of rotator cuff injuries. PMID- 12629429 TI - The effectiveness of worksite physical activity programs on physical activity, physical fitness, and health. AB - OBJECTIVE: To critically review the literature with respect to the effectiveness of worksite physical activity programs on physical activity, physical fitness, and health. DATA SOURCES: A search for relevant English-written papers published between 1980 and 2000 was conducted using MEDLINE, EMBASE, Sportdiscus, CINAHL, and Psychlit. The key words used involved a combination of concepts regarding type of study, study population, intervention, and outcome measure. In addition, a search was performed in our personal databases, as well as a reference search of the studies retrieved. STUDY SELECTION: The following criteria for inclusion were used: 1) randomized, controlled trial or nonrandomized, controlled trial; 2) working population; 3) worksite intervention program to promote employees' physical activity or physical fitness; and 4) physical activity, physical fitness, or health-related outcomes. DATA EXTRACTION: Two reviewers independently evaluated the quality of relevant studies using a predefined set of nine methodological criteria. Conclusions regarding the effectiveness of a worksite physical activity programs were based on a rating system consisting of five levels of evidence. DATA SYNTHESIS: Fifteen randomized, controlled trials and 11 nonrandomized, controlled trials met the criteria for inclusion and were reviewed. Six randomized, controlled trials and none of the nonrandomized, controlled trials were of high methodological quality. Strong evidence was found for a positive effect of a worksite physical activity program on physical activity and musculoskeletal disorders. Limited evidence was found for a positive effect on fatigue. For physical fitness, general health, blood serum lipids, and blood pressure, inconclusive or no evidence was found for a positive effect. CONCLUSIONS: To increase the level of physical activity and to reduce the risk of musculoskeletal disorders, we support implementation of worksite physical activity programs. For the other outcome measures, scientific evidence of the effectiveness of such a program is still limited or inconclusive, which is mainly the result of the small number of high-quality trials. Therefore, we recommend performing more randomized, controlled trials of high methodological quality, taking into account criteria such as randomization, blinding, and compliance. PMID- 12629430 TI - Stress fracture of the proximal phalanx of the second toe. PMID- 12629431 TI - Partial tear of the posterior deltoid muscle in an elderly woman. PMID- 12629432 TI - Anomalous intraarticular origin of the long head of biceps brachii. PMID- 12629442 TI - What's in a risk factor? "He who strikes the ball". AB - A major aim of epidemiology is to explore the prognosis and aetiology of diseases, in order to improve treatment and prevention. To avoid misleading interpretations of observed associations between a modifier and the occurrence of a disease, the all-purpose idiom "risk factor" should be replaced by 3 locutions with narrower meanings: a risk marker is a modifier which is associated (correlated) with disease prevalence (case-control studies) or incidence (cohort studies); a risk marker truly becomes a risk factor if its experimental correction (intervention studies) does improve the disease incidence or prognosis; a risk factor is promoted to the rank of cause if it is proved to be necessary (sine qua non) for the occurrence of the disease. A more rigorous vocabulary avoids premature claims for unsubstantiated treatments or preventions, and helps defining sound priorities in aetiological research. PMID- 12629443 TI - Relationship of regional adiposity to serum leptin level in nonobese Japanese type 2 diabetic male patients. AB - BACKGROUND: The aim of the present study was to investigate the relationships between serum leptin levels and regional adipose fat area, BMI, and the measures of variables including serum insulin in nonobese Japanese type 2 diabetic patients. METHODS: A total of 121 nonobese Japanese type 2 diabetic patients [aged 35 to 83 years, body mass index (BMI) (15.4 to 26.8 kg/m(2))] were studied. They all were male patients. In conjunction with serum leptin level, BMI, glycosylated hemoglobin (HbA(1c)), and fasting concentrations of plasma glucose and serum insulin and lipids (triglycerides, total and HDL cholesterol) were measured. RESULTS: Univariate regression analysis showed that serum leptin levels were positively correlated to subcutaneous (r=0.566, P<0.0001) and visceral (r=0.481, P<0.001) fat area in our diabetic patients. Furthermore, serum leptin levels were positively correlated to serum insulin (r=0.517, P<0.0001), BMI (r=0.428, P<0.0001), serum triglycerides (r=0.279, P<0.005), and age (r=0.225, P<0.05). There was, however, no relationship between serum leptin levels and measures of other variables including total and HDL cholesterol. Multiple regression analyses showed that serum leptin levels were predicted by subcutaneous fat area (F=5.92, P<0.0001) and serum insulin level (F=5.60, P<0.0001), which explained 29.0% of the variability of serum leptin concentrations in our nonobese Japanese type 2 diabetic male patients. Visceral fat area, BMI, serum triglycerides, and age, however, were not independently associated with serum leptin levels in our patients. CONCLUSIONS: These results indicate that serum leptin levels are reflective of subcutaneous fat area in nonobese Japanese type 2 diabetic male patients. PMID- 12629444 TI - Effects of a two-month rehabilitation program on substrate utilization during exercise in obese adolescents. AB - BACKGROUND: We assessed the impact of an exercise and diet program for two weeks in a specialized institute and six weeks at home on body composition and exercise metabolism in obese adolescents. METHODS: Fourteen obese adolescents took part in the two-week training course and seven continued exercise and diet at home. The substrate crossover point and the maximal fat oxidation point (Lipox max) were determined before and after the program, using indirect calorimetry. Individualized exercise training at Lipox max was programmed over the two months. RESULTS: At the end of the two-week program, all adolescents showed a decrease in weight (p<0.001). Only the seven subjects who continued exercise training at home showed improved exercise fat use (increased crossover point of 17% +/- 3 (p<0.005) and Lipox max of 12.5% +/- 1.5 (p<0.005)). CONCLUSION: The teenagers who had performed this targeted regular training for two months exhibit an increase in their ability to oxidize lipids at exercise. This increase is no longer found in those treated by diet alone. The efficiency of this targeted training protocol based on calorimetry for the treatment of teenager obesity will require longer studies on greater samples of subjects. PMID- 12629445 TI - An economic evaluation of Losartan therapy in type 2 diabetic patients with nephropathy: an analysis of the RENAAL study adapted to France. AB - BACKGROUND: The RENAAL study enrolled 1,513 patients with type 2 diabetes mellitus and nephropathy defined by the presence of proteinuria (urinary albumin: creatinine ratio 300 mg/g or proteinuria > 500 mg per day). Compared to placebo, losartan therapy reduced by 16% (p=0.02) the risk of a composite endpoint (doubling of baseline serum creatinine level, end stage renal disease, or death) and by 28% (p=0.002) the risk of progression to end stage renal disease (ESRD). METHODS: The objective of this study was to compare, using French economic data, the additional cost of losartan therapy with the savings in cost generated by a decrease in the number of end stage renal disease days. Prospectively collected health care resource utilization were used (N(losartan)=751, N(placebo)=762). The follow-up period was 4 years. RESULTS: The mean cumulative cost of losartan over 4 years was 1,603 euros per patient. The reduction in the number of ESRD days over 4 years in patients treated with losartan significantly decreased costs associated with ESRD by 7,438 euros per patient (CI 95%: 3,029 euros - 11,847 euros, p=0.001). Compared to the placebo group, the average cost per patient over 4 years in the losartan group was lower by 5,834 euros (CI 95%: 1,407 euros - 10,301 euros, p=0.01). CONCLUSION: In addition to the medical benefit, this analysis demonstrated the economic relevance of treatment with losartan in type 2 diabetic patients with nephropathy. PMID- 12629446 TI - Tolerance of Iloprost and results of treatment of chronic severe lower limb ischaemia in diabetic patients. A retrospective study of 64 consecutive cases. AB - BACKGROUND: The aim of this study was to assess the tolerance and report obtained results with a stable prostacyclin analogue (iloprost) in diabetic patients with severe forms of permanent lower limb ischaemia. METHODS: Sixty-four consecutive unselected patients, in stage III and IV of Leriche and Fontaine, turned down for vascular surgery after angiography and treated with iloprost during 28 days, were enrolled in this study. Patients were followed-up clinically (ischemic pain, trophic change, walking distance) and with transcutaneous oxymetry (D28). Long term assessment (6 and 12 months) was expressed as rate of death, major amputation and of live patients with viable limbs and walking. There was no manifestation of intolerance to iloprost. Were considered as responders patients offering a lack or significant decrease in pain, a reduction of trophic lesions and improvement or recovery of walking. RESULTS: Response at two months is lasting: 29 responders (45.3%) and 35 non-responders (54.7%). At 6 months and one year, we observed that 8 (12.5%) and 15 (24.1%) patients respectively had died; 19 (29.6%) and 22 (34.3%) patients underwent major amputation, but 41 (64%) and 34 (53.1%) patients were still alive with their limb and conservative walking. In responder group, at 6 months, 28 (96.5%) patients were alive without amputation for only 13 (37.1%) among non-responders. At one year, 79.3% of the responders and 31.4% of the non-responders were alive without amputation. A total loss of walking, a segmental amputation and a previous amputation of opposite limb were more often noted in no responder group. But no predictive factor was referred to TcPO(2) in particular. Results ware similar in the group of 136 non diabetic patients treated during the same period (67.9% alive with limb at 6 months). CONCLUSIONS: This retrospective study, despite its limitations, underlines the clinical particularities of critical ischaemia in diabetics and the good tolerance to iloprost. This point allowed patients, in non-surgical chronic critical ischaemia, to avoid being confined to bed and to access to benefits of a early physiotherapy, in association with local treatment. However, no predictive criterion of long-term results could be established, except initial clinical severity and clinical change one month after treatment. PMID- 12629447 TI - N(carboxymethyl)lysine as a biomarker for microvascular complications in type 2 diabetic patients. AB - AIMS: Hyperglycemia is linked to vascular dysfunction in patients with diabetes mellitus, either directly or through advanced glycation end product (AGE) formation. Experimental evidence has indicated the possible involvement of AGEs in the genesis of vascular complications. We investigated whether serum levels of AGEs and of the glycoxidation compound carboxymethyl-lysine (CML) were increased and correlated with vascular complications in type II diabetes mellitus. METHODS: Serum levels of AGEs and CML-human serum protein (CML-HSP) were measured by a specific immunoassay in 51 men and 26 women aged 58 +/- 6.1 years (mean +/- SD) who had been treated for type II diabetes mellitus for 11 +/- 8 years, and in a non-diabetic control group consisting of 39 men and 21 women aged 55.5 +/- 7.5 years. Patients with macroalbuminuria or abnormal creatinine clearance were excluded from the study. RESULTS: The serum levels of AGEs were significantly increased in patients with type II diabetes compared to controls (P<0.001). Blood levels of CML-HSP were significantly increased in diabetic patients compared to normal subjects [35.3 +/- 27.4 and 9.3 +/- 7.2 (mean +/- SD) pmol/mg of protein, respectively; P<0.0001]. In diabetic patients with retinopathy or microalbuminuria (urinary albumin excretion: UAE > 30 mg/24 h), CML-HSP levels were significantly higher (P<0.02), and even more elevated in patients with both complications. CONCLUSION: In patients with type II diabetes, CML-HSP levels that are at variance with the HbA(1c) index for blood glucose may be a biomarker of glycoxidation, and related to the development of microvascular complications. PMID- 12629448 TI - Functional intensified insulin therapy with short-acting insulin analog: effects on HbA1c and frequency of severe hypoglycemia. An observational cohort study. AB - OBJECTIVES: To assess the effect of a particular insulin regimen called "functional insulin therapy" using a short-acting insulin analog on the risk of severe hypoglycemia and the HbA(1c) level among patients already under intensive insulin therapy. DESIGN: A cohort of 110 patients with type 1 diabetes receiving intensive insulin therapy with regular insulin for several years was followed during one year after initiation of functional insulin therapy (FIT) with a short acting insulin analog. The glycemic control was assessed by the mean value of the last three HbA(1c) assays before the initiation of FIT and then by the mean of the following three. The number of severe hypoglycemic episodes/patient/year during the year preceding and the year following the initiation of FIT was recorded. RESULTS: The mean HbA(1c) level decreased on average by 0.7 percent during the 12-month study (p=0.0001) and the number of episodes of severe hypoglycemia fell to 75% of its previous level (p<0.05). CONCLUSION: Substitution of intensive insulin therapy using regular insulin for functional insulin therapy using short-acting insulin analog may improve glycemic control and reduce the risk of severe hypoglycemia. PMID- 12629450 TI - Relation of C-reactive protein to features of the metabolic syndrome in normal glucose tolerant, impaired glucose tolerant, and newly diagnosed type 2 diabetic subjects. AB - OBJECTIVE: To determine the relationship between CRP levels and the components of MS in normal glucose tolerant (NGT), impaired glucose tolerant (IGT), and Type 2 diabetic subjects. MATERIAL AND METHODS: A based cross-sectional population study was performed. Eligible subjects, men and non-pregnant women, 30 to 64 year of age, were randomly recruited. Subjects with acute or chronic diseases were excluded. Only newly diagnosed type 2 diabetic or hypertensive subjects were included. Disorders related to CRP increase, also were exclusion criteria. In accordance to WHO proposal, components of MS were: High Blood Pressure, Dyslipidemia, Obesity, and Microalbuminuria, and MS was defined, for the NGT, if at least two of the criteria were fulfilled and in addition the subject had insulin resistance. The MS in IGT and DM subjects was defined if at least two of the criteria were fulfilled. RESULTS: CRP was significantly associated with MS for the NGT (Odds ratio -OR- 3.8, CI(95%) 1.6-14.8), IGT (OR 4.9, CI(95%) 1.2 15.5), and diabetes (OR 5.6, CI(95%) 1.9-10.2). For NGT, after adjustment for obesity, CRP was not longer associated with MS. After adjust for obesity and fasting glucose (FG), the relationship between CRP and MS for IGT was lost. Finally, after adjustment for obesity, FG, and microalbuminuria, CRP was not longer associated with MS for diabetic subjects. CONCLUSIONS: This study show a significant relationship between CRP and MS which is maintained only by obesity in the NGT, by obesity and FG in the IGT, and by obesity, FG, and microalbuminuria in the newly diagnosed diabetic subjects. PMID- 12629449 TI - Management of hypertension and screening of renal complications by GPs in diabetic type 2 patients (France--2001). AB - BACKGROUND: Our aim was to assess the quality of the medical management by GPs of hypertension and renal insufficiency in type 2 diabetic patients. METHODS: A retrospective cohort study was run on a national random representative sample of 5,518 patients presenting with type 2 diabetes mellitus treated pharmacologically by a general practitioner from April 2000 to April 2001. RESULTS: Sixty percent of patients underwent a HbA(1c) measurement during the last 6 months and among them 27% exceeded the threshold of 8%. Glomerular Filtration Rate, calculated with the Cockcroft formula, was below 60 ml/min (confirmed renal failure) in 21.9% of patients and was in the 61-80 ml/min range (probable early renal insufficiency) in 27%. Proteinuria was documented in 30.1% of patients, 13.7% of whom were positive. Microalbuminuria was documented in 36%, 15% of whom were positive. Hypertension was treated pharmacologically in 59.6% of the sample (39.3% on monotherapy, 34.2% on double combination therapy and 26.5% on triple combination therapy or more). Blood pressure was >140 and/or 80 mmHg in 81.6% of treated patients and in 27% among untreated. CONCLUSION: These findings suggest that significant progress still needs to be made in the care and treatment of type 2 diabetic patients, especially those with hypertension, in order to reduce or delay the incidence of renal and cardiovascular complications. PMID- 12629451 TI - Application of "Concept Mapping" in obese subjects: a pilot study in normo and underreporters. AB - BACKGROUND: One important step in nutritional education consists in identifying the patient prior knowledge in order to better educate him. The objectives of this pilot study, on twelve obese patients (6 underreporting patients - UR - and 6 normoreporting patients - NR) were to point out advantages and limits of a method: "concept mapping", a graphic representation of the nature and organization of knowledge, and to show eventual cognitive differences between the two groups of patients. METHODS: Concept maps were drawed during interviews and analysed, considering the concepts, the links between them, their regrouping in knowledge fields. Conceps maps of UR and NR patients were compared. RESULTS: Our results confirm the advantages of concept mapping as the mean to explore the cognitive representation of patients. Some differences between UR and NR patients were readable on the maps: UR patients are more sensitive to the difficulties of the diet than the NR patients. UR patients tend to express more frequently their feelings or personal experiences, and to be more easily invaded by their feelings (more numerous links of personal expressions at UR patients). CONCLUSION: Concept mapping brings new information on knowledge of obese patients. They appear important to better know the cognitive representations of underreporting patients. With such information, it would be easier for healthcare providers to adapt educational programs to the patients' understanding, taking into account their misconceptions, errors and lacks of learning. PMID- 12629452 TI - Influence of oral antidiabetic drugs compliance on metabolic control in type 2 diabetes. A survey in general practice. AB - BACKGROUND: To address compliance with oral antidiabetic agents and its impact on metabolic control in type 2 diabetic patients treated in general practice. METHODS: Prospective assessment of self-reported compliance with a standardized questionnaire in an homogeneous cohort of 11,896 type 2 diabetic patients treated by their general practitioner with one or two oral antidiabetic agents, and analysis of determinants of compliance with treatment (age, diabetes duration, educational level, severity of complications, frequency of antidiabetic oral agents daily dosing). RESULTS: Optimal compliance (no omission) was reported in only 46% of cases. In multivariate analysis, HbA(1c) levels were positively correlated with age, daily dosing frequency of oral antidiabetic agents and low educational level, but not with diabetes duration. Low daily dosing was associated with a better metabolic control. HbA(1c) levels were associated with compliance with treatment, with a 1.4% mean difference between group with optimal and group with worst compliance. CONCLUSIONS: These results suggest the interest of reducing daily dosing frequency of oral antidiabetic agents, in order to improve compliance with treatment and metabolic control. PMID- 12629453 TI - Post-partum recurrent sarcoidosis associated with type 1 diabetes mellitus. AB - Immunologic abnormalities observed in sarcoidosis may suggest a link between this affection and autoimmune endocrine diseases. Indeed a high frequency of autoimmune thyroid diseases is observed in sarcoidosis. However association of type 1 insulin-dependent diabetes mellitus with sarcoidosis is rare. We report the case of a type 1 diabetic woman in whom clinical and biological signs of sarcoidosis appeared after her first pregnancy with a relapse in the post-partum period of a second pregnancy. Diagnosis of sarcoidosis was established on characteristic cutaneous, articular and pulmonary manifestations associated with elevated plasma levels of angiotensin converting enzyme. From this case, association between type 1 diabetes mellitus and sarcoidosis has been discussed as well as reciprocal relationship between sarcoidosis and pregnancy. Since familial history of sarcoidosis was present in this case, familial aspects of sarcoidosis have also been reviewed. PMID- 12629454 TI - A new dark side of the Maillard's products. PMID- 12629455 TI - [Gastroplasty: complications and their prevention]. AB - Bariatric surgery is now frequently proposed for the treatment of morbid or complicated obesity since the introduction of minimally invasive laparoscopic anti-obesity operations such as the adjustable silicone gastric binding gastroplasty. However this reversible procedure in not always as safe as presumed and the results in weight loss may be sometimes disappointing. Side effects are common and early or late complications occured in more than 20% out of the patients. They are favoured by post operative eating disorders. Nutritional consequences are probably underestimated and are not limited to uncomfortable digestive symptoms. Some deficiencies in micronutriments have been described. The worsening of previous eating disorders or psychosocial abnormalities are not seldom. Gastroplasty is not an harmless procedure. A good selection in patients, a regular follow up, nutritional advices and psychosocial management by a multidisciplinar team are required to reduce complications after gastroplasty. PMID- 12629456 TI - The use of near infrared spectroscopy in sports medicine. AB - In the last 15 years the study of the human muscle energetics in sports medicine underwent a radical change thanks to the progressive introduction of non-invasive techniques, including near infrared (NIR) spectroscopy (NIRS). NIR light (700 1000 nm) penetrates skin, subcutaneous fat and underlying muscle, and is either absorbed (by oxy- and deoxy-haemoglobin) or scattered within the tissue. NIRS is a non-invasive and relatively low cost optical technique that is becoming a widely used instrument for measuring muscle O(2) saturation and changes in haemoglobin volume. Muscle O(2) saturation represents a dynamic balance between O(2) supply and O(2) consumption in the small vessels such as the capillary, arteriolar and venular bed. NIRS offers the advantage of being less restrictive than (31)P-magnetic resonance spectroscopy with regard to muscle performance and more comfortable and suitable for the monitoring, with high temporal resolution (up to 10 Hz), of multiple muscle groups. The aim of this review is to summarise the NIRS instrumentation and the measurable parameters, the role of NIRS in muscle exercise physiology, and the applications in sports medicine. The advantages and the problems of NIRS measurements, in resting and exercising skeletal muscles, are reported. The results of several studies suggest that NIRS is a powerful tool for being applied successfully in sports medicine. NIRS can objectively evaluate muscle oxidative metabolism in athletes and its modifications following potential therapeutic strategies and specific training programs. PMID- 12629457 TI - Test performance indicators from a single soccer specific fitness test differentiate between highly trained and recreationally active soccer players. AB - AIM: The aim of this study was to investigate whether a single soccer specific fitness test (SSFT) could differentiate between highly trained and recreationally active soccer players in selected test performance indicators. METHODS: SUBJECTS: 13 Academy Scholars (AS) from a professional soccer club and 10 Recreational Players (RP) agreed to participate in this study. Test 1--VO(2) max was estimated from a progressive shuttle run test to exhaustion. Test 2--The SSFT was controlled by an automated procedure and alternated between walking, sprinting, jogging and cruise running speeds. Three activity blocks (1A, 2A and 3A) were separated by 3 min rest periods in which blood lactate samples were drawn. The 3 blocks of activity (Part A) were followed by 10 min of exercise at speeds alternating between jogging and cruise running (Part B). RESULTS: Estimated VO(2) max did not significantly differ between groups, although a trend for a higher aerobic capacity was evident in AS (p<0.09). Exercising heart rates did not differ between AS and RP, however, recovery heart rates taken from the 3 min rest periods were significantly lower in AS compared with RP following blocks 1A (124.65 b x min(-1) +/-7.73 and 133.98 b x min(-1) +/-6.63), (p<0.05) and 3A (129.91 b x min(-1) +/-10.21 and 138.85 b x min(-1) +/-8.70), (p<0.01). Blood lactate concentrations were significantly elevated in AS in comparison to RP following blocks 2A (6.91 mmol x l(-1) +/-2.67 and 4.74 mmol x l(-1) +/-1.28) and 3A (7.18 mmol x l(-1) +/-2.97 and 4.88 mmol x l(-1) +/-1.50), (p<0.05). AS sustained significantly faster average sprint times in block 3A compared with RP (3.18 sec +/-0.12 and 3.31 sec +/-0.12), (p<0.05). CONCLUSION: The results of this study show that highly trained soccer players are able to sustain, and more quickly recover from, high intensity intermittent exercise. PMID- 12629458 TI - Effects of running, static stretching and practice jumps on explosive force production and jumping performance. AB - AIM: The interaction between running, stretching and practice jumps during warm up for jumping tests has not been investigated. The purpose of the present study was to compare the effects of running, static stretching of the leg extensors and practice jumps on explosive force production and jumping performance. METHODS: Sixteen volunteers (13 male and 3 female) participated in five different warm-ups in a randomised order prior to the performance of two jumping tests. The warm-ups were control, 4 min run, static stretch, run + stretch, and run + stretch + practice jumps. After a 2 min rest, a concentric jump and a drop jump were performed, which yielded 6 variables expressing fast force production and jumping performance of the leg extensor muscles (concentric jump height, peak force, rate of force developed, drop jump height, contact time and height/time). RESULTS: Generally the stretching warm-up produced the lowest values and the run or run + stretch + jumps warm-ups produced the highest values of explosive force production. There were no significant differences (p<0.05) between the control and run + stretch warm-ups, whereas the run yielded significantly better scores than the run + stretch warm-up for drop jump height (3.2%), concentric jump height (3.4%) and peak concentric force (2.7%) and rate of force developed (15.4%). CONCLUSION: The results indicated that submaximum running and practice jumps had a positive effect whereas static stretching had a negative influence on explosive force and jumping performance. It was suggested that an alternative for static stretching should be considered in warm-ups prior to power activities. PMID- 12629459 TI - Reliability and gender differences of static explosive grip parameters based on force-time curves. AB - AIM: The purpose of this study was to examine the reliability and gender differences of static explosive grip (SEG) parameters based on force-time curves. METHODS: EXPERIMENTAL DESIGN: data were collected from 2 trials of SEG tests on the same day and 2 trials 1 week later. PARTICIPANTS: 50 healthy young males (21.2+/-1.57 yrs) and females (20.1+/-1.18 yrs) participated in this study. MEASURES: 12 variables based on factors such as time-course of force, average force, integrated area, maximal rate of force development, the product of force and inclination and exponential function of force development (EXP-M) were selected as force-time parameters in SEG testing. RESULTS: A significant reliability was obtained for all force-time parameters except the time to 90% of maximal grip strength (MAX), with the reliability of integrated area, average force, maximal rate of force development (DFmax), and the product of force and inclination being particularly high (ICC=0.633-0.930 for the same day, ICC=0.612 0.828 for different days). There were gender differences for all parameters except the time to MAX and EXP-M (p<0.05). In addition, many of the correlation coefficients between parameters were high, particularly for males. The average force, integrated area for 1 sec, and the product of force and inclination during the developmental phase significantly correlated with MAX in both males and females. The ratio between males and females in these force-time parameters was smaller than that of MAX (71.3%). CONCLUSION: It is considered that these parameters are dependent on the speed of muscle contraction. These results suggest that the evaluation parameters, such as average force, integrated area, the product of force and inclination, and maximal rate of force development are useful for evaluating static explosive grip. PMID- 12629460 TI - Effects of submaximal intensity cycle ergometry for one hour on substrate utilisation in trained prepubertal boys versus trained adults. AB - AIM: Substrate utilisation during exercise might be different in trained prepubertal boys compared to trained adults, so dietary advice usually given to endurance trained adults may need to be adapted for endurance trained children. METHODS: EXPERIMENTAL DESIGN: subjects pedalled for 1 hour on an ergocycle at 40% or 60% of Wmax (maximal aerobic power). PARTICIPANTS: 14 boys and 13 adults active in endurance sport (swimming). MEASURES: pulmonary respiratory gas exchange ratio [RER = VCO(2)/VO(2)], used as the equivalent of the non protein respiratory quotient (RQ), was measured at rest, at the 15(th), 30(th), 45(thv and 60(th) min of exercise in order to calculate energy expenditure. RESULTS: Relative resting energy expenditure was significantly higher in boys than in adults. During exercise, energy expenditure (EE) was significantly lower in children than in adults (p<0.001), whereas fat-free mass relative EE was only influenced by intensity. Relative EE from carbohydrates (EE(CHO)) was lower in children compared to adults, even if the highest intensity involves a higher EE(CHO) in both groups (p<0.05). Boys oxidised more lipid at 40% of Wmax than 60% of Wmax or adults at either intensity. CHO utilisation was significantly increased at 60% vs 40% of Wmax, yet lower in boys than in men (p<0.01). Lipids' use, in g x min(-1) x kg-1)FFM, was significantly higher in boys than in men at 40% of Wmax (p<0.01). CONCLUSION: Trained children, at rest, had greater fat-free mass energy expenditure than adults, which may be linked to growth processes that remain unclear. During exercise, in boys, the main observation was the increase in CHO dependence relative with the intensity. Thus, it appears that CHO feeding is as important in endurance-trained boys as in endurance-trained adults, especially when exercise intensity approaches that of competition or training sessions. PMID- 12629461 TI - Pulmonary function during cycling and running in triathletes. AB - AIM: Running performance has become key to the triathlete's overall performance. We still know relatively little about the factors that define the ability to perform a good run after cycling, however, and the perception of discomfort during the first minutes of this post-cycling running has yet to be satisfactorily explained. Pulmonary volumes (i.e., residual volume, RV, and functional residual capacity, FRC) have been demonstrated to be impaired after a cycle-run succession in triathletes but not after a run-run succession that is matched in terms of intensity and duration. Cycling in itself and/or the succession of two different exercises (i.e., cycling and running) may explain this phenomenon, but the exact mechanism has not yet been determined. METHODS: Thirteen young male triathletes participated in three different exercise trials: 30 min of cycling followed by 20 min of running (C-R), 30 min of control cycling (C) and 20 min of control running (R). Pulmonary volumes and flows were measured 10 min before and 10 min after each trial. During all trials, ventilatory data were collected every minute using an automated breath-by-breath system. RESULTS: The results showed that 1) C induced significant increases in RV, FRC and RV/TLC (2.31+/-0.18 vs 2.01+/-0.17 L, 4.35+/-0.24 vs 4.01+/-0.25 L, and 27.21+/-1.62 vs 23.98+/-1.55, respectively, after versus before C) and 2) there were no significant pulmonary volume or flow changes after C-R or R. CONCLUSION: We concluded that 1) cycling exercise in itself seems to increase the post-exercise pulmonary volume changes which could lead to respiratory muscle alterations and 2) one likely explanation for this finding appears to be the crouched position of cycling. PMID- 12629462 TI - Influence of the base-line determination on work efficiency during submaximal cycling. AB - AIM: The purpose of this study was to compare work efficiency values (WE = work accomplished/energy expenditure above exercising with 0 load) among different unloaded base-line correction techniques for different power outputs. METHODS: Twelve healthy men performed 6 5-min steady-state exercises of 0 (unloaded), 40, 80, 120, 160 and 200 W at a pedalling rate of 90 rpm on a cycle ergometer. Three different unloaded base-line corrections were used for WE calculation: an actual measurement of VO(2) corresponding to the unloaded pedalling exercise, the y intercept value of the linear regression between VO(2) and power output and the y intercept value of the curvilinear VO(2)-power regression. RESULTS: The present study demonstrated that WE was significantly higher when determined using the actual measurement of the unloaded VO(2) than y-intercept values of the linear (p<0.001) and curvilinear (p<0.05) VO(2)-power regressions. WE based on theoretical determinations (linear vs curvilinear regressions) were not significantly different. The power output significantly affected all WE index, with higher WE being measured when the power output was low and lower WE when power output was high. CONCLUSION: The high WE values determined using the actual VO(2) measurement could be explained by (i) the additional energy expended to stabilise the body in addition to the energy expenditure of moving the lower limbs without power production and by (ii) the difficulty to experimentally reproduce the unloaded condition. The large range of WE values measured in the present study is due to differences in the procedures used to determine the unloaded VO(2) (and thus differences in unloaded VO(2) values) as well as differences in the cycling intensities. PMID- 12629463 TI - Early effects of short-term aerobic training. Physiological responses to graded exercise. AB - AIM: The aim of this study was to find out how early the moderate training effects appear and to check the hypothesis that familiarization with exercise protocol may contribute to an early physiological responses to training in previously sedentary subjects. METHODS: Twelve male, sedentary volunteers (22.0+/ 0.7 yrs) were submitted to 3 weeks of a bicycle ergometer training, consisting of 45 min of exercise (at 70% VO(2)max), 3-4 times a week. The subjects performed 4 incremental exercise tests until volitional exhaustion: 2 before training (C1 and C2), and then after 1 (T1) and 3 (T3) weeks of training. During exercise HR, VO(2), electrical activity (EMG) of rectus femoris, biceps femoris, soleus and trapezius muscles were recorded and blood samples were taken for blood lactate (LA) determination. RESULTS: Already after 1 week of training HR decreased (p<0.05) with a further decline after 3 weeks the training (p<0.01). Maximal work load after 3 weeks of training increased to 277+/-10.4 W vs 250+/-9.5 W (p<0.05), VO(2)max achieved higher values than in C1 and C2 tests (p<0.05) and LA and EMG thresholds were elevated (p<0.05). CONCLUSION: A decrease in the resting and submaximal heart rate is the earliest effect of increased physical activity. Familiarization to exercise protocol decreased EMG of biceps femoris and soleus muscles during exercise, but did not influence that of rectus femoris muscle the most engaged during cycling. PMID- 12629464 TI - Effects of beta-hydroxy-beta-methylbutyrate and creatine monohydrate supplementation on the aerobic and anaerobic capacity of highly trained athletes. AB - AIM: The aim of this study was to investigate the effects of 6 wks oral supplementation of beta-hydroxy-beta-methylbutyrate (HMB) and a mixture of HMB and creatine monohydrate (HMBCr) on aerobic and anaerobic capacity in highly trained athletes. It was hypothesised that HMB and HMBCr would have positive effects on aerobic and anaerobic power. METHODS: A prospective study involving a repeated measures design was utilised where subjects underwent testing prior to, and immediately after, a 6 wks supplementation period. Elite, male rugby league players (n=27) were divided into 3 groups, a control group (n=6), a HMB group (3 g/d; n=10) and a HMBCr group (3 g/d HMB + 3 g/d Cr; n=11). Testing involved a multistage fitness test to determine aerobic power and a 60 sec maximal cycle test to determine anaerobic capacity. Peak power, total work and peak lactate levels were measured in the anaerobic cycle test. RESULTS: Two-way repeated measures ANOVA revealed no effect of HMB or HMBCr on any of the measured parameters in comparison to the control group. CONCLUSION: Aerobic and anaerobic ability of highly trained male athletes is unaffected by 6 wks oral supplementation with HMB or a combination of HMB and creatine monohydrate. PMID- 12629465 TI - Dynamics of muscle strength improvement during isokinetic rehabilitation of athletes with ACL rupture and chondromalacia patellae. AB - AIM: To assess quantitatively dynamics and extent of the increase in muscle strength during isokinetic rehabilitation. METHODS: EXPERIMENTAL DESIGN: daily measurements of muscle strength; detailed testing at the beginning and at the end of rehabilitation. SETTING: Cybex Rehabilitation Center, Zagreb. SUBJECTS: 44 athletes (31 m, 13 F, age 16-35), 3 injury-defined groups: athletes with ACL rupture (non-reconstructed and reconstructed) and chondromalacia patellae. INTERVENTIONS: all subjects underwent isokinetic rehabilitation on Cybex Orthotron KT2 device, using individually designed protocols (extension and flexion exercises, concentric muscle contractions, 15 treatments). MEASUREMENTS: monitoring of daily progress on rehabilitation device and detailed testing on diagnostic device. RESULTS: All patients showed considerable improvement. Muscle strength improved on average 141% (SD=110) in ACL-reconstructed group, 144% (SD=130) for chondromalacia patellae group and 150% (SD=74) for ACL-non reconstructed group, comparing to initial strength. Dynamic status tested on Cybex Otrhotron diagnostic device prior and after rehabilitation strongly correlated with final progress monitored on the rehabilitation device. CONCLUSION: Isokinetic rehabilitation is a quick and effective method in treating knee injuries in athletes. Both types of objective criteria have shown significant increase in muscle strength. The improvement of muscle strength was on the average 149% (SD=101), which is about 10% daily for 15 treatments. The greatest progress, 19% per day, occurred during first five days. The athletes were able to resume their sport activities as follows: patients from chondromalacia patellae group, and most of them from the non-reconstructed ACL group were back in competition within a month, while 75% from the ACL reconstructed group came back within 3 months, and the rest of them within 5 months. PMID- 12629467 TI - Sex differences in delayed onset muscle soreness. AB - AIM: There is agreement that females report greater pain in response to typical experimental pain stimuli than males. However, investigations of sex differences in the sensation of delayed onset muscle soreness (DOMS) have equivocal RESULTS: The objective of this investigation was to examine sex differences in the pain from DOMS with an adequate sample size, quantification of stimulus intensity, and 2 measures of pain. METHODS: Sixty-seven participants (52% females) completed a 2 session protocol. DOMS was induced using eccentric resistance exercises in the elbow flexors of the non-dominant arm. The intensity of the eccentric contractions was based upon concentric strength. Pain response was measured 48 hrs later. The dependent variables were pressure threshold, which was assessed using a dolorimeter, and pain intensity when the arm was moved through full active range of motion, which was assessed with a visual analog scale. RESULTS: The occurrence of DOMS was confirmed by a decrease in pressure threshold after the eccentric contractions and higher pain intensity in the arm that performed the eccentric contractions than the arm that did not. Females reported lower pain intensities (M=3.41, SD=2.13) compared to males (M=5.12, SD=2.05), but no significant sex difference was found in pressure threshold. CONCLUSION: In this investigation, females reported lower muscle pain intensity than males, but showed no sex difference in pressure threshold. These and previous findings suggest that the detection of a sex difference in muscle pain depends upon the methodology of inducing DOMS and measuring sensation. PMID- 12629466 TI - Myositis ossificans as a complication of a muscle tendon junction strain of long head of biceps. A case report. AB - An unusual case of myositis ossificans secondary to a muscle tendon junction (MTJ) strain of long head of biceps in a young athlete is reported. Plain radiographs, ultrasonography and MRI in association with clinical assessment showed the appearance and the evolution of this pathological entity. This case had a resolution of pain and function after 3 months of conservative treatment. At 6 months follow-up, the athlete became asymptomatic and he gradually returned back to his sports activity. PMID- 12629468 TI - Age and gender-related physical activity. A descriptive study in children using accelerometry. AB - AIM: Precise measures of habitual physical activity are necessary in studies designed to: 1) document the frequency and distribution of physical activity in defined population groups; 2) determine the amount or dose of physical activity requires to influence specific health parameters. The purpose of this study was to document the age and gender-related physical activity levels on a sample of school children. METHODS: The sample for the present study comprised 157 children (boys n=64 and girls n=93), aged 8 to 15 years-old. The CSA activity monitor was used as an objective measure of daily physical activity. Each student in the present study was scheduled to wear the CSA 3 times during the week of monitoring. RESULTS: Boys were involved (p=/<0.05) in more time MVPA than girls. However only in the 11-13 year old group were found significant differences (p=/<0.05). Within gender, significant differences were found out among 11-13 years old (48.7 min) and 14-16 years (72.2 min). The time in MVPA increased across age group in both males and females. Boys participated in more periods of continuous physical activity, bouts of 10 and 20 min respectively, than girls. However no significant differences were found out. CONCLUSION: Our data showed that children under observation, excepted girls in the 11-13 year old group, appear to meet the minimum physical activity level recommended for health. The data also suggested that boys are more active than girls are and that they were significantly more engaged in more time MVPA than girls did. Surprisingly our data showed an increase in MVPA time as age increases. Further studies are needed to compare the different cut off points assessed during daily activity. PMID- 12629469 TI - Secular trends in physical performance of Australian children. Evidence from the Talent Search program. AB - AIM: This study examined trends in aerobic fitness in 12-15-year-old South Australian schoolchildren in the years 1995-2000, based on data from the Australian Sports Commission's Talent Search program. METHODS: A total of 18,631 children were tested. The aerobic performance test used was the 20 m shuttle run test (20 mSRT). The 20 mSRT scores were expressed as completed laps, and converted to estimated VO(2)max values. RESULTS: There were significant declines (p=0.04-0.0001) across all age-gender slices, equivalent to 0.18 to 0.36 ml O(2) x kg(-1) x min(-1) x yr(-1), or about 0.4-0.8% of mean values per year. The rate of decline is consistent with several other Australian and overseas studies in the years 1980-2000, which have used a variety of aerobic tests across a wide range of age groups. In relation to children of similar age in 7 other countries, Australian children show poor to average aerobic fitness levels. CONCLUSION: It is possible that the decay in Australian children's aerobic fitness is in part due to reduced physical activity. PMID- 12629470 TI - Modulation of immune responses by treadmill exercise in Sprague-Dawley rats. AB - AIM: The duration-dependence of the effect of forced treadmill exercise on the immune system is a subject of ongoing research. In this study, the effect of forced treadmill exercise on immune responses was investigated by evaluating the lymphocyte subset fractions in the peripheral blood and spleen of Sprague-Dawley rats. METHODS: EXPERIMENTAL DESIGN: Comparative investigation over 8 weeks. SETTING: Experimental animal laboratory. PARTICIPANTS: Male Sprague-Dawley rats 5 weeks of age, weighing 150+/-10 g. INTERVENTIONS: Animals were randomly assigned to one of the 4 following groups: the control group, the 1-week-exercise group, the 4-week-exercise group, and the 8-week-exercise group. MEASURES: Lymphocyte subset fractions, including those for T, B, CD4+, and CD8+ cells and the T/B and CD4+/CD8+ ratios in the peripheral blood and spleen were measured via flow cytometric analysis after treadmill exercise. RESULTS: The T cell and CD4+ cell fractions in both the peripheral blood and spleen were increased significantly after 8 weeks of treadmill exercise, but the B cell and CD8+ cell fractions did not change significantly. CONCLUSION: From the results of the present study, it is suggested that a period of one week is insufficient to eliminate the effects of exercise-induced stress, that 4 weeks are needed to return to the control state, and that at least 8 weeks are needed in order for exercise of moderate intensity to have a positive effect on the immune system. PMID- 12629471 TI - The effect of acute plasma volume expansion on venous capacitance. AB - AIM: The purpose of this study was to examine the effect of plasma volume (PV) expansion, induced by 1 single bout of high intensity exercise, on venous capacitance. METHODS: Fourteen active but untrained healthy young males were randomly assigned to an exercise (n=7) or control group (n=7). The exercise protocol consisted of 8 4-min bouts of exercise at 70-85% of maximal oxygen consumption (VO(2)max) with 5-min recovery periods between bouts. Resting and exercise HR and venous capacitance were assessed before and 24 hours after one bout of cycle exercise. Exercise HR was assessed during cycle exercise at 30, 45, and 60% of VO(2)max, whereas forearm and leg venous capacitance were examined by determining limb volume change after 2 min of venous occlusion at 50 mmHg. PV, plasma albumin, and sodium and potassium concentration were analysed pre and 24 hours post exercise. RESULTS: Twenty-four hours after one single bout of exercise, PV was increased by 7.7%. The increase in PV was accompanied by a decrease in exercise HR of 5.8% at the 2 higher intensities (45 and 60% of VO(2)max) but no change in venous capacitance was found. No significant changes in plasma albumin and sodium concentration occurred, although potassium concentration was significantly increased by 29%. CONCLUSION: An acute exercise induced increase in plasma volume appears to be able to reduce exercise HR but has no effect on venous capacitance. PMID- 12629472 TI - Over-the-counter drug use amongst athletes and non-athletes. AB - AIM: Many over-the-counter (OTC) drugs used in the symptomatic relief of upper respiratory tract (URT) conditions are banned by sports governing bodies. It would appear therefore that athletes are being penalised for practising conventional pharmacological methods in the management of common ailments. The aim was to identify any differences between athletes and non-athletes and amongst athletic groups, with respect to the prevalence of URT conditions and the use of OTC drugs to treat such conditions. METHODS: Questionnaires were distributed at domestic and international athletics meetings and at university lectures and tutorials. Respondents (n=401) represented both track and field athletes (n=199) and non-athletes (n=202). RESULTS: No differences were found between athletes and non-athletes and between elite and non-elite athletes in terms of the frequency of episodes of URT conditions reported in the previous year. A higher proportion of elite, as opposed to non-elite athletes did not take OTC medicines (p=0.028) and of those that did take OTC medicines a higher proportion of elite athletes (68%) as opposed to non-elite (32%) took those not containing sympathomimetics, banned by the International Olympic Committee (IOC). Athletes were found to have greater knowledge of IOC banned OTC drugs (p=0.002) and within this group, elite athletes were most knowledgeable (p=0.0003). Although most respondents (81%) believed that OTC drugs should not be prohibited in sport, athletes made up the greatest proportion in support of prohibition (23.5% as opposed to 14.4% of non athletes) with elite as opposed to non-elite most in favour (p=0.0181). CONCLUSION: These results suggest that URT conditions are no more prevalent between athletes and non-athletes or between endurance and power athletes. Athletes competing at the highest level tended to avoid OTC medicines or those containing IOC banned drugs and were most knowledgeable in terms of banned OTC drugs and most in favour of their prohibition suggesting that the control mechanisms in place are only reaching elite athletes. PMID- 12629473 TI - [Vascular and endovascular surgery from the history of techniques, surgical indications]. PMID- 12629474 TI - [Criteria of evidence-based medicine for correct surgical indication]. PMID- 12629475 TI - [Indications for reconstructive and endovascular therapy of abdominal aortic aneurysms]. PMID- 12629476 TI - [Surgical indications in abdominal-thoracic aneurysms]. PMID- 12629477 TI - [Ischemic heart disease in patients with arteriopathies]. PMID- 12629478 TI - [Indications for associated renal revascularization]. PMID- 12629479 TI - [When claudication becomes surgical]. PMID- 12629480 TI - [Indications for treatment of carotid stenosis. Choice of loco-regional anesthesia and general anesthesia]. PMID- 12629481 TI - [Social relevance of venous lymphatic diseases]. PMID- 12629482 TI - [Importance of the CEAP classification]. PMID- 12629483 TI - [Diagnostic course in patients with phlebopathies]. PMID- 12629484 TI - [Chronic venous insufficiency: surgical therapy based on hemodynamic damage]. PMID- 12629485 TI - [Correction of venous hypertension in venous dysplasias]. PMID- 12629486 TI - [Indications for surgery of the lymphatics]. PMID- 12629487 TI - Diurnal regulation of arylalkylamine N-acetyltransferase activity in chicken retinal cells in vitro: analysis of culture conditions. AB - PURPOSE: Arylalkylamine N-acetyltransferase (AANAT) is a key regulatory enzyme in the synthesis of melatonin, which displays daily fluctuations in chicken retinal photoreceptors in vivo. The purpose of the present study was to determine if cultures of embryonic neural retina cells express diurnal rhythms of AANAT activity. METHODS: Cell cultures were prepared from chick embryonic day 6 neural retina and incubated for 4 to 8 days in vitro (DIV). Cells were incubated under a daily light-dark (LD) cycle and were harvested day and night. Culture conditions were modified to test the effects of cell density, serum concentration, incubation temperature, S-(4-nitrobenzyl)-6-thioinosine (NBTI), and taurine on AANAT activity. AANAT activity was assayed in cell homogenates by measuring the catalytic formation of N-acetyltryptamine from tryptamine and acetyl coenzyme A. RESULTS: Cells cultured in medium containing 10% fetal bovine serum (FBS) failed to show any diurnal fluctuation in AANAT activity on DIV 5 and 6. However, if the culture medium was replaced on DIV 4 with one containing 1% FBS, and 5 microM NBTI or 5 mM taurine, the cells expressed significant diurnal rhythms of enzyme activity. NBTI was more potent and effective than taurine. Culture conditions were optimized with respect to cell density, serum concentration, incubation temperature, and NBTI concentration. Under optimized conditions, overall cell survival and the density of photoreceptor cells were increased relative to that with the other culture conditions tested. CONCLUSIONS: The results indicate that diurnal rhythms of AANAT activity are expressed in embryonic retinal cells incubated under particular culture conditions. The results show that the mechanisms regulating melatonin synthesis in chicken retinal cells are established during early embryonic life. This culture preparation will be useful in elucidating the photic control mechanisms involved in regulation of melatonin biosynthesis in photoreceptor cells. PMID- 12629489 TI - Wounds: looking beyond the surface. PMID- 12629493 TI - Perceived and objective risk in children of patients with peripheral arterial disease. AB - This study examined perceived and objective health risks, health promotive behavior, risk perception, and knowledge of risk factors for peripheral arterial disease (PAD) in children of patients with PAD. Children of patients who had lower extremity distal arterial reconstructive surgery or amputation for complications of PAD completed an investigator-developed questionnaire. Risk factor and behavioral measures were self-reported. Data were collected by telephone interview. The sample consisted of 15 children of 6 patients. Subjects displayed optimistic bias regarding their risk for developing PAD, with 67% reporting their parent's illness had no impact on their health behaviors. Fifty three percent felt their comparative risk was "about the same" as same-age, same sex peers. Risk factor knowledge varied and none mentioned hypertension, age, obesity, or gender; the most commonly cited risk factor was diabetes mellitus. Forty-seven percent of the offspring never smoked, less than half exercised regularly, most were overweight or had class I or class II obesity, and most were unaware of their blood pressure or cholesterol levels. Despite frequent interactions with their affected parent, these children exhibited poor understanding of risk factors and personal risk of developing PAD and most did not participate in health promotive behaviors. Educational efforts by national societies promoting an understanding of modifiable risk factors must be improved. Future studies need to explore interventions designed to improve risk perception and health promotive behaviors. PMID- 12629492 TI - Treating patients with peripheral arterial disease and claudication. AB - The vascular nurse plays an important role in the treatment of patients with peripheral arterial disease (PAD), a prevalent atherosclerotic occlusive disease that affects approximately 8 to 12 million people in the United States. Approximately 4 to 5 million individuals with PAD experience claudication, the exercise-induced ischemic pain in the lower extremities that is relieved upon rest. Both PAD and claudication are associated with increased morbidity and mortality, limitations in functional capacity, and a decreased quality of life. Despite its prevalence, PAD is often undiagnosed and, therefore, increases the risk for cardiovascular ischemic events, disease progression, functional disability, amputation, and death. Risk factors for PAD and claudication are similar to those for other atherosclerotic diseases, including age, cigarette smoking, diabetes mellitus, hypertension, dyslipidemia, and hyperhomocysteinemia. Effective treatment to normalize these risk factors can reduce disease progression and the incidence of cardiovascular ischemic events. Claudication symptoms can be improved most effectively through exercise training, which may be used in conjunction with medications specifically indicated to improve these symptoms. Vascular nurses, practicing in a multitude of inpatient and outpatient settings, can assist patients with risk-factor modifications and behavioral changes to help them stop smoking, maintain glycemic control, normalize high blood pressure and lipid levels, and ensure initiation of lifelong antiplatelet therapy and participation in exercise rehabilitation programs, thus, promoting positive outcomes for patients with claudication. PMID- 12629495 TI - Hyperhomocysteinemia and vascular disease. PMID- 12629494 TI - Patient with compartment syndrome of the lower extremity. PMID- 12629496 TI - Help is on the way...but we need to work together. PMID- 12629497 TI - Finding focus through consensus. PMID- 12629498 TI - Informed consent in pharmacogenomics. AB - The Pharmacogenomics Journal (2002) 2, 343. doi:10.1038/sj.tpj.6500151 PMID- 12629500 TI - Antidepressants and gene expression profiling: how to SNARE novel drug targets. PMID- 12629501 TI - Patent or perish? An ethical approach to patenting human genes and proteins. PMID- 12629502 TI - A complicated relationship: fulfilling the interactive needs of the T lymphocyte and the dendritic cell. AB - T cells recognize antigenic peptides displayed on the surface of MHC-bearing antigen-presenting cells (APCs), and with sufficient costimulation become activated. However, the ability of an APC (even bearing the correct peptide) to initiate and fulfill the requirements for T cell activation is not easily achieved. Naive T cells use multiple copies of a single receptor to survey the vast array of peptides presented on an APC, and require multiple receptor engagements to initiate T cell activation. Dendritic cells (DCs) are specialized cells with optimal capabilities for priming naive CD4+ T cells. Activation occurs, after initial antigen recognition by T cells, followed by a rapid dialogue between the T cells and the DCs. The resulting changes in both the cytoskeleton and the expression or regulation of cell-surface molecules on both cell types act to further strengthen engagement. In this report, we review the fundamentals of CD4+ T helper cell : DC interactions and discuss recent data concerning the molecular characteristics of this engagement. PMID- 12629503 TI - Differential expression of VAMP2/synaptobrevin-2 after antidepressant and electroconvulsive treatment in rat frontal cortex. AB - The biological basis for the therapeutic mechanisms of depression is still unknown. We have previously performed expressed-sequence tag (EST) analysis to identify some molecular machinery responsible for antidepressant effect. Then, we developed our original cDNA microarray, on which cDNA fragments identified as antidepressant-related genes/ESTs were spotted. In this study, with this microarray followed by Western blot analysis, we have demonstrated the induction of vesicle-associated membrane protein 2(VAMP2/synaptobrevin-2) in rat frontal cortex not only after chronic antidepressant treatment, but also after repeated electroconvulsive treatment. On the other hand, expression of SNAP-25 and syntaxin-1 was not changed by these treatments. These components make a soluble N ethylmaleimide-sensitive fusion protein attachment protein receptor complex with VAMP2 and mediate the synaptic vesicle docking/fusion machinery. In conclusion, it is suggested that VAMP2/synaptobrevin-2 plays important roles in the antidepressant effects. Our results may contribute to a novel model for the therapeutic mechanism of depression and new molecular targets for the development of therapeutic agents. PMID- 12629504 TI - Pharmacogenomic analysis of rhIL-11 treatment in the HLA-B27 rat model of inflammatory bowel disease. AB - Recombinant human interleukin-11 (rhIL-11) reduces the clinical signs and histological lesions of inflammatory bowel disease (IBD) in transgenic rats expressing the human major histocompatability complex (MHC) class I allele, HLA B27. To elucidate the pharmacogenomic effects of rhIL-11 in this model, we examined the global gene expression pattern in inflamed colonic tissue before and following rhIL-11 treatment using oligonucleotide microarrays. In total, 175 disease-related genes were identified. Increased expression of genes involved in antigen presentation, cell death and inflammation, and decreased expression of metabolic genes was associated with disease. A total of 27 disease-related genes returned to normal expression levels following rhIL-11 treatment including the MHC class II gene RT1-DMbeta. rhIL-11 induced the expression of four intestinal epithelial growth factors. These gene expression patterns indicate that treatment of inflammatory bowel disease with rhIL-11 affects class II antigen processing and colonic epithelial cell proliferation and metabolism. PMID- 12629506 TI - Interpretation of genetic association studies in complex disease. PMID- 12629505 TI - Association between CYP2D6 genotype and tardive dyskinesia in Korean schizophrenics. AB - The CYP2D6 gene codes for human cytochrome P450 2D6 enzyme, which is responsible for the metabolism of many psychiatric drugs. In schizophrenic patients treated with neuroleptics, decreased or loss of function CYP2D6 alleles may contribute to the development of tardive dyskinesia (TD), a movement disorder that frequently occurs with chronic neuroleptic treatment. The goal of this study was to determine whether the occurrence of TD is associated with CYP2D6 genotype in a cohort of Korean schizophrenics by employing a CYP450 GeneChip((R)) oligonucleotide microarray and PCR assays to screen for 19 CYP2D6 alleles. Our results revealed that males with at least one decreased or loss of function allele have a moderately greater chance of developing TD than males with only wild-type alleles. Female schizophrenics did not have a significantly greater chance of developing TD. Our results demonstrate the utility of CYP2D6 microarrays to assess genotype status in this Korean cohort. PMID- 12629507 TI - Preferential induction of necrosis in human breast cancer cells by a p53 peptide derived from the MDM2 binding site. AB - p53 is the most frequently altered gene in human cancer and therefore represents an ideal target for cancer therapy. Several amino terminal p53-derived synthetic peptides were tested for their antiproliferative effects on breast cancer cell lines MDA-MB-468 (mutant p53), MCF-7 (overexpressed wild-type p53), and MDA-MB 157 (null p53). p53(15)Ant peptide representing the majority of the mouse double minute clone 2 binding site on p53 (amino acids 12-26) fused to the Drosophila carrier protein Antennapedia was the most effective. p53(15)Ant peptide induced rapid, nonapoptotic cell death resembling necrosis in all breast cancer cells; however, minimal cytotoxicity was observed in the nonmalignant breast epithelial cells MCF-10-2A and MCF-10F. Bioinformatic/biophysical analysis utilizing hydrophobic moment and secondary structure predictions as well as circular dichroism spectroscopy revealed an alpha-helical hydrophobic peptide structure with membrane disruptive potential. Based on these findings, p53(15)Ant peptide may be a novel peptide cancer therapeutic because it induces necrotic cell death and not apoptosis, which is uncommon in traditional cancer therapy. PMID- 12629508 TI - Identification of novel pRb binding sites using CpG microarrays suggests that E2F recruits pRb to specific genomic sites during S phase. AB - The retinoblastoma (Rb) tumor suppressor protein is an important regulator of cell proliferation and differentiation. Many studies have shown that pRb can negatively regulate the activity of the E2F family of transcription factors during G(0) and G(1) phases of the cell cycle, perhaps by serving as a bridge between the E2Fs and transcriptional repressors such as histone deacetylases and methylases. However, pRb has also been shown to localize to discrete DNA foci during S phase, a time at which pRb is thought to be dissociated from E2F. Numerous other DNA binding proteins have been shown to interact with pRb, suggesting that pRb may control progression through S phase by binding to sites in the genome distinct from E2F target gene promoters. To test this hypothesis, we have identified novel pRb binding sites within the human genome using an unbiased approach which relies upon a combination of chromatin immunoprecipitation and CpG microarray analysis. To provide the greatest opportunity of finding distinct sets of pRb binding sites, we examined pRb binding in chromatin obtained from human Raji cells synchronized in either G(0)/G(1) phase or S phase. These experiments have allowed us to identify a large set of new genomic binding sites for the pRb protein. We found that some sites are occupied by pRb only during G(0)/G(1) phase, as would be predicted from previous models of pRb function. We also identified sites to which pRb bound only during S phase and other sites which were bound constitutively by pRb. Surprisingly, we found that E2F1 was present at most of the CpG islands bound by pRb, independent of the phase of the cell cycle. Thus, although pRb has the potential to interact with numerous transcription factors, our data suggest that the majority of DNA-bound pRb is recruited to E2F target promoters during both G(0)/G(1) and S phases. PMID- 12629509 TI - The structural determinants responsible for c-Fos protein proteasomal degradation differ according to the conditions of expression. AB - c-fos gene is expressed constitutively in a number of tissues as well as in certain tumor cells and is inducible, in general rapidly and transiently, in virtually all other cell types by a variety of stimuli. Its protein product, c Fos, is a short-lived transcription factor that heterodimerizes with various protein partners within the AP-1 transcription complex via leucine zipper/leucine zipper interactions for binding to specific DNA sequences. It is mostly, if not exclusively, degraded by the proteasome. To localize the determinant(s) responsible for its instability, we have conducted a genetic analysis in which the half-lives of c-Fos mutants and chimeras made with the stable EGFP reporter protein were compared under two experimental conditions taken as example of continous and inducible expression. Those were constitutive expression in asynchronously growing Balb/C 3T3 mouse embryo fibroblasts and transient induction in the same cells undergoing the G0/G1 phase transition upon stimulation by serum. Our work shows that c-Fos is degraded faster in synchronous than in asynchronous cells. This difference in turnover is primarily accounted for by several mechanisms. First, in asynchronous cells, a unique C-terminal destabilizer is active whereas, in serum-stimulated cells two destabilizers located at both extremities of the protein are functional. Second, heterodimerization and/or binding to DNA accelerates protein degradation only during the G0/G1 phase transition. Adding another level of complexity to turnover control, phosphorylation at serines 362 and 374, which are c-Fos phosphorylation sites largely modified during the G0/G1 phase transition, stabilizes c-Fos much more efficiently in asynchronous than in serum-stimulated cells. In both cases, the reduced degradation rate is due to inhibition of the activity of the C terminal destabilizer. However, in serum-stimulated cells, this effect is partially masked by the activation of the N-terminal destabilizer and basic domain/leucine zipper-dependent mechanisms. Taken together, our data show that multiple degradation mechanisms, differing according to the conditions of expression, may operate on c-Fos to ensure a proper level and/or timing of expression. Moreover, they also indicate that the half-life of c-Fos during the G0/G1 phase transition is determined by a delicate balance between opposing stabilizing and destabilizing mechanisms operating at the same time. PMID- 12629510 TI - Annexin II expression is reduced or lost in prostate cancer cells and its re expression inhibits prostate cancer cell migration. AB - While studying Bim, a BH3-only proapoptotic protein, we identified an approximately 36 kDa protein, which was abundantly expressed in all five strains of primary normal human prostate (NHP) epithelial cells but significantly reduced or lost in seven prostate cancer cell lines. The approximately 36 kDa protein was subsequently identified as annexin II by proteomic approach and confirmed by Western blotting using an annexin II-specific antibody. Conventional and 2D SDS PAGE, together with Western blotting, also revealed reduced or lost expression of annexin I in prostate cancer cells. Subcellular localization studies revealed that in NHP cells, annexin II was distributed both in the cytosol and underneath the plasma membrane, but not on the cell surface. Prostate cancer cells showed reduced levels as well as altered expression patterns of annexin II. Since annexins play important roles in maintaining Ca(2+) homeostasis and regulating the cytoskeleton and cell motility, we hypothesized that the reduced or lost expression of annexin I/II might promote certain aggressive phenotypes of prostate cancer cells. In subsequent experiments, we indeed observed that restoration of annexin II expression inhibited the migration of the transfected prostate cancer cells without affecting cell proliferation or apoptosis. Hence, our results suggest that annexin II, and, likely, annexin I, may be endogenous suppressors of prostate cancer cell migration and their reduced or lost expression may contribute to prostate cancer development and progression. PMID- 12629511 TI - Alterations of anaphase-promoting complex genes in human colon cancer cells. AB - Ubiquitin-mediated proteolysis of cell cycle regulators is a major element of the cell cycle control. The anaphase-promoting complex (APC/C) is a large multisubunit ubiquitin-protein ligase required for the ubiquitination and degradation of G1 and mitotic checkpoint regulators. APC/C-dependent proteolysis regulates cyclin levels in G1, and triggers the separation of sister chromatids at the metaphase-anaphase transition and the destruction of mitotic cyclins at the end of mitosis. Furthermore, it was recently shown that APC/C regulates the degradation of crucial regulators of signal transduction pathways. We report here gene alterations in several components of this complex in human colon cancer cells, including APC6/CDC16 and APC8/CDC23 which are known to be key function elements. The experimental expression of a truncation mutant of APC8/CDC23 subunit (CDC23DeltaTPR) leads to abnormal levels of APC/C targets such as cyclin B1 and disturbs the cell cycle progression of colon epithelial cells through mitosis. Overall, these data support the hypothesis of a deleterious role of these mutations during colorectal carcinogenesis. PMID- 12629512 TI - Werner's syndrome protein is phosphorylated in an ATR/ATM-dependent manner following replication arrest and DNA damage induced during the S phase of the cell cycle. AB - Werner's syndrome (WS) is an autosomal recessive disorder, characterized at the cellular level by genomic instability in the form of variegated translocation mosaicism and extensive deletions. Individuals with WS prematurely develop multiple age-related pathologies and exhibit increased incidence of cancer. WRN, the gene defective in WS, encodes a 160-kDa protein (WRN), which has 3' 5'exonuclease, DNA helicase and DNA-dependent ATPase activities. WRN-defective cells are hypersensitive to certain genotoxic agents that cause replication arrest and/or double-strand breaks at the replication fork, suggesting a pivotal role for WRN in the protection of the integrity of the genoma during the DNA replication process. Here, we show that WRN is phosphorylated through an ATR/ATM dependent pathway in response to replication blockage. However, we provide evidence that WRN phosphorylation is not essential for its subnuclear relocalization after replication arrest. Finally, we show that WRN and ATR colocalize after replication fork arrest, suggesting that WRN and the ATR kinase collaborate to prevent genome instability during the S phase. PMID- 12629513 TI - PDGF-D is a potent transforming and angiogenic growth factor. AB - Platelet-derived growth factors (PDGFs) are important for normal tissue growth and maintenance. Overexpression of the classical PDGFs, PDGF-A and PDGF-B, has been linked to several diseases, including cancer, fibrotic disease and atherosclerosis. Recently, two novel PDGFs, PDGF-C and PDGF-D, were discovered. It has not yet been established whether PDGF-C and PDGF-D are linked to disease phenotypes like the classical PDGFs. PDGF-B, the cellular homologue of the viral simian sarcoma oncogene v-sis, is known to potently induce cellular transformation through activation of PDGF receptor (PDGFR)-beta. In this work, we have determined the transformation efficacy of PDGF-D in comparison with that of PDGF-C and PDGF-B. PDGF-D is a potent transforming growth factor for NIH/3T3 cells, and the transformed cells displayed stress fibre reorganization, increased proliferation rate, anchorage-independent growth in soft agar, ability to induce tumours in nude mice, and upregulation of vascular endothelial growth factor. Morphological analyses of the vasculatures from the PDGF-isoform-expressing tumours revealed marked differences suggesting differential signalling through the two PDGF receptors in tumour vessel development and remodelling. In summary, these results suggest that PDGF-D induce cellular transformation and promote tumour growth by accelerating the proliferation rate of the tumour cells, and by stimulation of tumour neovascularization. PMID- 12629514 TI - Presenilin-dependent gamma-secretase activity mediates the intramembranous cleavage of CD44. AB - CD44 is the major adhesion molecule for the extracellular matrix components and is implicated in a wide variety of physiological and pathological processes including the regulation of tumor cell growth and metastasis. Our previous studies have shown that CD44 undergoes sequential proteolytic cleavages in the extracellular and transmembrane domains and the cleavage product derived from CD44 intramembranous cleavage acts as a signal transduction molecule. However, the underlying mechanism of the intramembranous cleavage of CD44 remains to be elucidated. In the present study, we report for the first time that CD44 is a substrate of the presenilin (PS)-dependent gamma-secretase. We demonstrate that the intramembranous cleavage of CD44 induced by 12-O-tetradecanoylphorbol 13 acetate (TPA) treatment or mechanical scraping is blocked by gamma-secretase inhibitors in U251MG cells and that this cleavage is also inhibited in PS deficient mouse embryonic fibroblasts. Furthermore, we showed that PS1 is redistributed to ruffling areas of the plasma membrane similarly to CD44 after TPA treatment, supporting our biochemical observation that PS1 is involved in the intramembranous cleavage of CD44. Our present findings suggest important implications for understanding CD44-dependent signal transduction and a potential role of PS/gamma-secretase activity in the functional regulation of adhesion molecules. PMID- 12629515 TI - Interleukin-6 promotes cervical tumor growth by VEGF-dependent angiogenesis via a STAT3 pathway. AB - Interleukin-6 (IL-6) has received particular attention in the pathogenesis of cervical cancer, although the underlying mechanism remains elusive. This study revealed that IL-6 promotes in vivo tumor growth of human cervical cancer C33A cells, but does not substantially alter their in vitro growth kinetics. The in vivo angiogenic assays showed that IL-6 increases angiogenic activity in human cervical cancer cells, an effect that is specifically associated with upregulation of vascular endothelial growth factor (VEGF). Also, using anti-VEGF antibody to block VEGF function significantly inhibited IL-6-mediated angiogenesis and tumor growth in nude mice, strongly supporting the critical role of VEGF in the IL-6-mediated cervical tumorigenesis. Accordingly, the signaling pathway downstream of IL-6/IL-6R responsible for the regulation of VEGF was investigated. Notably, pharmacological inhibition of PI3-K or MAPK failed to inhibit IL-6-mediated transcriptional upregulation of VEGF. Meanwhile, blocking STAT3 pathway with dominant-negative mutant STAT3D effectively abolished IL-6 induced VEGF mRNA. In transient transfections, a luciferase reporter construct containing the full-length 1.5-kb VEGF promoter or a 1.2-kb fragment lacking the known hypoxic-response element also exhibited the same degree of response to IL 6. Additionally, transient transfection of STAT3D downregulated the 1.2-kb VEGF promoter luciferase reporter stimulated by IL-6. Based on the above phenomenon combined with the concomitant increased tumor expression of IL-6 and VEGF in cervical cancer tissues, we conclude that IL-6 may promote cervical tumorigenesis by activating VEGF-mediated angiogenesis via a STAT3 pathway. PMID- 12629516 TI - Quantitation of HERV-K env gene expression and splicing in human breast cancer. AB - Human endogenous retroviruses (HERVs) comprise up to 8% of the human genome. In previous studies, we demonstrated that type 1 HERV-K envelope (env) transcripts are expressed in most human breast cancers, but not in normal breast tissues. In the current study, we report that type 2 HERV-K env transcripts are also present in human breast cancers. By real-time RT-PCR, the expression of HERV-K env transcripts was 5-10-fold higher in breast cancer cell lines treated with estradiol and progesterone than in cells without treatment, and expression was significantly higher in most breast cancer tissues than in normal breast tissues. Furthermore, both types of HERV-K env transcripts were capable of being spliced into subgenomic env transcripts and various splice donor and acceptor sites were detected in breast cancers. The selective expression and distribution of multiple HERV-K endogenous retroviral element splice variants in breast cancer, but not in normal controls, suggests that they are novel breast tumor markers. PMID- 12629517 TI - Wnt signaling in B-cell neoplasia. AB - Wnts comprise a family of secreted proteins that interact with receptors consisting of a Frizzled (Fz) family member alone or complexed with LDL receptor related proteins (LRP5/6). Wnt signaling plays a crucial role in both development and differentiation, and activation of a 'canonical' Wnt pathway resulting in beta-catenin stabilization is associated with several types of human cancers. To date, little is known about potential Wnt signaling in mature lymphocytes or lymphoid neoplasia. Herein, we have analysed Wnt signaling in mature B cells (lymphomas) and plasma cells (multiple myeloma). Both Fz and LRP5/6 mRNAs were expressed in myeloma lines, but LRP5/6 were not observed in lymphomas. In myelomas, a canonical Wnt signaling pathway was activated following treatment with Wnt-3a as assessed by accumulation of beta-catenin, but beta-catenin levels actually decreased in lymphoma cells. Wnt-3a treatment further led to striking morphological changes in myeloma cells accompanied by rearrangement of the actin cytoskeleton. Morphological changes were associated with a second Wnt pathway dependent on Rho activation. These results suggest that Wnt responsiveness is a stage-specific phenomenon in B-cell development and that the morphological changes associated with Wnt signaling may play a role in the motility and metastatic potential of myeloma cells. PMID- 12629518 TI - Gab1 is required for EGF receptor signaling and the transformation by activated ErbB2. AB - Grb2-associated binder-1 (Gab1) is a pleckstrin homology (PH) domain-containing adapter molecule that is believed to function downstream of receptors for growth factors and cytokines. We previously found that deficiency in the mouse Gab1 gene led to embryonic lethality and defects in ERK activation in response to growth factors and cytokines. Here, we established immortalized Gab1-/- cell lines and analysed roles of Gab1 in growth factor-mediated signaling and oncogenesis. EGF dependent activation of c-Raf and Mek1/2, which function upstream of ERKs, was perturbed in Gab1-/- cells. EGF-mediated upregulation of GTP-bound form of Ras was also reduced in these cells. EGF-dependent GTP/GDP exchange activity for Ras was suppressed in the Gab1-/- cells and expression of a constitutively active Sos restored ERK activation in these cells, indicating that Gab1 functions upstream of Ras. Furthermore, activated form of ErbB2 (active ErbB2)-mediated transformation, such as colony formation in soft agar and tumor formation in nude mice, was strongly suppressed when the Gab1-/- cells were transfected with active ErbB2, whereas the active Sos efficiently induced transformation of Gab1-/- cells. The data show that Gab1 plays an essential role in EGF-receptor/ErbB mediated cell proliferation and oncogenesis. PMID- 12629520 TI - Patterns of expression of chromosome 17 genes in primary cultures of normal ovarian surface epithelia and epithelial ovarian cancer cell lines. AB - Oligonucleotide microarray analysis was applied to assess the expression profile of 332 probe sets representing 308 genes or expressed sequence tags (ESTs) that map to chromosome 17 in order to address epigenetic events that result in alterations in gene expression in epithelial ovarian cancer (EOC). Expression profiles were generated from 12 primary cultures derived from normal ovarian surface epithelium (NOSE) and four long-term cultures (TOV-81D, TOV-112D, TOV-21G and OV-90) derived from EOCs that have been previously characterized and shown to mimic features of the tumoral cells from which they were derived. The expression values of all 332 probe sets is highly correlated across the 12 NOSEs (89% correlation coefficients >0.90). In two-way comparisons, differential patterns of gene expression were observed for 157 probe sets for which the expression value of at least one EOC cell line fell outside the limits of the range of expression of the 12 NOSEs. When compared to NOSEs, four genes displayed similar differential patterns of gene expression across all four EOC cell lines, and 26 genes displayed similar differential patterns of gene expression across the three EOC cell lines (TOV-112D, TOV-21G and OV-90) representing tumoral cells derived from the most aggressive disease. A total of 17 genes displayed differential patterns of gene expression greater than threefold in at least one EOC cell line in comparison to NOSE, and three genes were differentially expressed greater than threefold across all aggressive cell lines. The analysis of a selected number of genes by RT-PCR revealed patterns of gene expression comparable to those observed by microarray analysis in the majority of samples tested. Comparison of expression profiles of differentially expressed genes identified by microarray analysis in two-way comparisons of the EOC cell lines and the NOSEs with published reports revealed 10 genes previously implicated in ovarian tumorigenesis and 18 in tumorigenesis of other types of cancer. The differential pattern of gene expression of genes that map to both the p and q arm of chromosome 17 is consistent with the hypothesis that a number of genes that map to this chromosome are implicated in the etiology of ovarian cancer. PMID- 12629519 TI - Generation and functional characterization of intracellular antibodies interacting with the kinase domain of human EGF receptor. AB - Intracellular expression of single-chain antibodies (scFvs) represents a promising approach for selective interference with cellular proto-oncogenes such as the epidermal growth factor receptor (EGFR). Previously, we have shown that intrabodies targeted to the lumen of the endoplasmic reticulum prevent the transit of EGFR or the related ErbB2 molecule to the cell surface, thereby inactivating their transforming potential. While intramolecular disulfide bridges important for antibody stability are correctly formed during expression in the secretory pathway, scFvs expressed in the reducing environment of the cytosol are often inactive. To overcome this problem and to generate antibody fragments that interact with the intracellular domain of human EGFR in the cytoplasm, here we have chosen a two-step approach combining classical selection of scFvs by phage display with subsequent expression in yeast. After enrichment of EGFR-specific antibody fragments from a combinatorial library by biopanning, a yeast two-hybrid screen was performed using the intracellular domain of EGFR as bait. Screening of 1.5 x 10(5) preselected scFv plasmids under highly stringent conditions yielded 223 colonies that represented at least five independent scFv clones functional in the intracellular milieu of eukaryotic cells. Interaction of selected antibody fragments with the intracellular domain of EGFR was confirmed in GST pull-down and coimmunoprecipitation experiments. Upon cytoplasmic expression in human tumor cells, scFvs colocalized with EGFR at the plasma membrane demonstrating their functionality in vivo. PMID- 12629521 TI - Epigenetic inactivation of the candidate 3p21.3 suppressor gene BLU in human cancers. AB - Many distinct regions of 3p show frequent allelic losses in a wide range of tumour types. Previously, the BLU candidate tumour suppressor gene (TSG) encoded by a gene-rich critical deleted region in 3p21.3 was found to be inactivated rarely in lung cancer, although expression was downregulated in a subset of lung tumour cell lines. To elucidate the role of BLU in tumorigenesis, we analysed BLU promoter methylation status in tumour cell lines and detected promoter region hypermethylation in 39% lung, 42% breast, 50% kidney, 86% neuroblastoma and 80% nasopharyngeal (NPC) tumour cell lines. Methylation of the BLU promoter region correlated with the downregulation of BLU transcript expression in tumour cell lines. Expression was recovered in tumour cell lines treated with 5-aza 2 deoxycytidine. Exogenous expression of BLU in neuroblastoma (SK-N-SH) and NSCLC (NCI-H1299) resulted in reduced colony formation efficiency, in vitro. Furthermore, methylation of the BLU promoter region was detected in primary sporadic SCLC (14%), NSCLC (19%) and neuroblastoma (41%). As frequent methylation of the RASSF1A 3p21.3 TSG has also been reported in these tumour types, we investigated whether BLU and RASSF1A methylation were independent or related events. No correlation was found between hypermethylation of RASSF1A and BLU promoter region CpG islands in SCLC or neuroblastoma. However, there was association between RASSF1A and BLU methylation in NSCLC (P=0.0031). Our data suggest that in SCLC and neuroblastoma, RASSF1A and BLU methylations are unrelated events and not a manifestation of a regional alteration in epigenetic status, while in NSCLC there may be a regional methylation effect. Together, these data suggest a significant role for epigenetic inactivation of BLU in the pathogenesis of common human cancers and that methylation inactivation of BLU occurs independent of RASSF1A in SCLC and neuroblastoma tumours. PMID- 12629522 TI - Control of 5-hydroxytryptamine release in the dorsal raphe nucleus by the noradrenergic system in rat brain. Role of alpha-adrenoceptors. AB - The interactions between the brainstem serotonergic (5-hydroxytryptamine, 5-HT) and noradrenergic (NA) systems are important for the pathophysiology and treatment of affective disorders. We examined the influence of alpha adrenoceptors on 5-HT and NA release in the rat dorsal raphe nucleus (DR) using microdialysis. 5-HT and NA concentrations in DR dialysates were virtually suppressed by TTX and increased by veratridine. The local and systemic administration of the alpha(1)-adrenoceptor antagonist prazosin reduced the DR 5 HT output but not that of NA. The maximal 5-HT reduction induced by local prazosin administration (-78% at 100 microM) was more marked than by its systemic administration (-43% at 0.3 mg/kg). The local application of NA and desipramine, to increase the tone on DR alpha(1)-adrenoceptors, did not enhance 5-HT release. The local (100 microM) or systemic (0.1-1 mg/kg s.c.) administration of clonidine reduced 5-HT and NA release (-48 and -79%, respectively, at 1 mg/kg), an effect reversed by RX-821002, which by itself increased both amines when given systemically. DSP-4 pretreatment prevented the effects of clonidine on 5-HT, suggesting the participation of alpha(2)-adrenoceptors on NA elements. Moreover, the systemic effect of clonidine on 5-HT (but not NA) was cancelled by lesion of the lateral habenula and by anesthesia, and was slightly enhanced by cortical transection. These data support the view that alpha(1)-adrenoceptors in the DR tonically stimulate 5-HT release, possibly at nearly maximal tone. Likewise, the 5-HT release is modulated by alpha(2)-adrenoceptors in NA neurons and in forebrain areas involved in the distal control of 5-HT neurons. PMID- 12629523 TI - Stimulation of P2Y1 receptors causes anxiolytic-like effects in the rat elevated plus-maze: implications for the involvement of P2Y1 receptor-mediated nitric oxide production. AB - The widespread and abundant distribution of P2Y receptors in the mammalian brain suggests important functions for these receptors in the CNS. To study a possible involvement of the P2Y receptors in the regulation of fear and anxiety, the influences of the P2Y(1,11,12) receptor-specific agonist adenosine 5'-O-(2 thiodiphosphate) (ADPbetaS), the P2X(1,3) receptor agonist alpha,beta-methylene ATP (alpha,betameATP), the unspecific P2 receptor antagonist pyridoxalphosphate-6 azopheny l-2',4'-disulfonic acid (PPADS), and the specific P2Y(1) receptor antagonist N(6)-methyl-2'-deoxyadenosine-3',5'-bisphosphate (MRS 2179) on the elevated plus-maze behavior of the rat were investigated. All tested compounds were given intracerebroventricularly (0.5 microl). ADPbetaS (50 and 500 fmol) produced an anxiolytic-like behavioral profile reflected by an increase of the open arm exploration. The anxiolytic-like effects were antagonized by pretreatment with PPADS (5 pmol) or MRS 2179 (5 pmol). Both compounds caused anxiogenic-like effects when given alone. Furthermore, the anxiolytic-like effects of ADPbetaS could be antagonized by pretreatment with the nitric oxide synthase (NOS) inhibitor N(w)-nitro-L-arginine methyl ester (L-NAME). In addition, the anxiogenic-like effects of PPADS were reversed by the pretreatment with L-arginine (500 pmol), which is the natural substrate for NOS, but not by D arginine (500 pmol), which is not. Immunofluorescence staining revealed the presence of P2Y(1) receptors on neurons in different brain regions such as hypothalamus, amygdala, hippocampus and the periaqueductal gray. Furthermore, the colocalization of P2Y(1) receptors and neuronal NOS (nNOS) on some neurons in these regions could be demonstrated. The highest density of P2Y(1)- and nNOS immunoreactivity was detected in the dorsomedial hypothalamic nucleus. Taken together, the present results suggest that P2Y(1) receptors are involved in the modulation of anxiety in the rat. The anxiolytic-like effects after stimulation of P2Y(1) receptors seem to be in close connection with the related nitric oxide production. PMID- 12629524 TI - Modulation of serotonergic function in rat brain by VN2222, a serotonin reuptake inhibitor and 5-HT1A receptor agonist. AB - VN2222 (1-(benzo[b]thiophen-3-yl)-3-[4-(2-methoxiphenyl piperazin-1-yl]propan-1 ol) is a potential antidepressant with high affinity for the serotonin transporter and 5-HT(1A) receptors. Locally applied, VN2222 enhanced the extracellular 5-hydroxytryptamine (5-HT) concentration (5-HT(ext)) in rat striatum to 780% of baseline whereas its systemic administration (1-10 mg/kg s.c.) reduced 5-HT(ext). In the presence of citalopram, 8-OH-DPAT or VN2222 applied in medial prefrontal cortex reduced 5-HT(ext). Fluoxetine, VN2222, and 8 OH-DPAT suppressed the firing rate of dorsal raphe 5-HT neurons (ED(50): 790, 14.9, and 0.8 microg/kg i.v., respectively). These effects were antagonized by WAY 100635. Administration of VN2222 for 2 weeks desensitized 5-HT(1A) receptors as assessed by microdialysis and single-unit recordings (ED(50) values for 8-OH DPAT were 0.45 and 2.34 microg/kg i.v. for controls and rats treated with 6 mg/kg day VN2222). These results show that VN2222 is a mixed 5-HT reuptake inhibitor/5 HT(1A) agonist that markedly desensitizes 5-HT(1A) autoreceptors. These properties suggest that it may be a clinically effective dual action antidepressant drug. PMID- 12629525 TI - Morphine tolerance and reward but not expression of morphine dependence are inhibited by the selective glutamate carboxypeptidase II (GCP II, NAALADase) inhibitor, 2-PMPA. AB - Inhibition of glutamate carboxypeptidase II (GCP II; NAALADase) produces a variety of effects on glutamatergic neurotransmission. The aim of this study was to investigate effects of GCP II inhibition with the selective inhibitor, 2-PMPA, on: (a) development of tolerance to the antinociceptive effects, (b) withdrawal, and (c) conditioned reward produced by morphine in C57/Bl mice. The degree of tolerance was assessed using the tail-flick test before and after 6 days of twice daily (b.i.d.) administration of 2-PMPA and 10 mg/kg of morphine. Opioid withdrawal was measured 3 days after twice daily morphine (30 or 10 mg/kg) administration, followed by naloxone challenge. Conditioned morphine reward was investigated using conditioned place preference with a single morphine dose (10 mg/kg). High doses of 2-PMPA inhibited the development of morphine tolerance (resembling the effect of 7.5 mg/kg of the NMDA receptor antagonist, memantine) while not affecting the severity of withdrawal. A high dose of 2-PMPA (100 mg/kg) also significantly potentiated morphine withdrawal, but inhibited both acquisition and expression of morphine-induced conditioned place preference. Memantine inhibited the intensity of morphine withdrawal as well as acquisition and expression of morphine-induced conditioned place preference. In addition, 2 PMPA did not affect learning or memory retrieval in a simple two-trial test, nor did it produce withdrawal symptoms in morphine-dependent, placebo-challenged mice. Results suggest involvement of GCP II (NAALADase) in phenomena related to opioid addiction. PMID- 12629526 TI - Expression of amphetamine-induced behavioral sensitization after short- and long term withdrawal periods: participation of mu- and delta-opioid receptors. AB - Repeated amphetamine administration results in behavioral sensitization, an enduring behavioral transformation expressed after short and long periods of withdrawal. To investigate the participation of the opioid system in amphetamine induced behavioral sensitization, we studied the effect of naloxone, an opioid receptor antagonist, on the expression of behavioral sensitization tested after short- (2 days) and long-term (14 days) withdrawal periods. In addition, using quantitative competitive RT-PCR, we examined the levels of mu-opioid receptor (MOR) and delta-opioid receptor (DOR) mRNA in the nucleus accumbens shell (NAcSh) and ventral tegmental area (VTA) of behaviorally sensitized rats, at these two withdrawal times. This study showed that whereas naloxone did not modify the expression of behavioral sensitization tested after 2 days of withdrawal, it completely blocked the expression when tested after 14 days of withdrawal. DOR and MOR mRNA levels were not modified in the NAcSh of rats expressing behavioral sensitization after 2 or 14 days of withdrawal. Conversely, DOR and MOR mRNA levels were elevated in the VTA of animals expressing behavioral sensitization after 2 days of withdrawal. However, whereas DOR mRNA returned to control levels, MOR mRNA levels remained elevated in animals expressing behavioral sensitization after 14 days of withdrawal. These results indicate a striking difference between the role played by opioid receptors in the expression of amphetamine-induced behavioral sensitization, when tested after short- or long-term withdrawal periods. In addition, our results support the notion that repeated amphetamine induced changes in opioid receptor expression may contribute to the perpetuation of psychostimulant abuse and/or relapse. PMID- 12629527 TI - Effects of cocaine on c-fos and NGFI-B mRNA expression in transgenic mice underexpressing glucocorticoid receptors. AB - Numerous evidences suggest that stress and stress-related hormones can modulate the activity of the brain reward pathway and thus may account for individual vulnerability towards the reinforcing effects of drugs of abuse. Transgenic (TG) mice expressing an antisense mRNA against the glucocorticoid receptor (GR), which partially blocks GR expression, were used to assess the role of GR dysfunction on cocaine (COC)-induced c-fos and Nerve-Growth Factor Inducible-B (NGFI-B, or Nur77) gene expression. These two genes belong to different families of transcription factors and have been shown to be modulated by various dopaminergic drugs. TG and wild-type (WT) mice were both acutely and repeatedly treated with COC (20 mg/kg, i.p.). In the chronic experiment, mice received a 5-day treatment of COC and were challenged 5 days later with COC or vehicle. Locomotor activity was assessed during the entire chronic experiment in the mouse home cages. Animals were sacrificed 1 h after the last injection and NGFI-B and c-fos mRNA levels in the prefrontal cortex, the nucleus accumbens and the striatum were measured by in situ hybridization. Acute COC administration led to significantly smaller c-fos increases in TG mice compared to WT, whereas repeated COC treatment potentiated c-fos induction both in TG and WT mice to equivalent levels. TG mice displayed higher basal NGFI-B expression in the nucleus accumbens and the level of NGFI-B mRNA was differently modulated by COC in TG mice compared to WT mice. In accordance with data on c-fos expression, behavioral data indicate a blunted locomotor effect on the first COC injection in TG mice, a phenomenon corrected by the repeated COC treatment. These results suggest that an alteration of the hypothalamus-pituitary-adrenal axis can modify COC-induced regulation of the transcription factors c-fos and NGFI-B, and that these changes parallel those seen at the behavioral level. It also demonstrates that the differences at the behavioral and molecular levels noted between TG and WT mice after acute COC injection disappear following repeated COC administration, suggesting that repeated COC has a greater impact in TG mice underexpressing GRs. PMID- 12629528 TI - Prefrontocortical dopamine loss in rats delays long-term extinction of contextual conditioned fear, and reduces social interaction without affecting short-term social interaction memory. AB - Prefrontal dopamine loss delays extinction of cued fear conditioning responses, but its role in contextual fear conditioning has not been explored. Medial prefrontal lesions also enhance social interaction in rats, but the role of prefrontal dopamine loss on social interaction memory is not known. Besides, a role for subcortical accumbal dopamine on mnesic changes after prefrontal dopamine manipulation has been proposed but not explored. The objective was to study the involvement of dopaminergic neurotransmission in the medial prefrontal cortex (mPFC) and nucleus accumbens in two mnesic tasks: contextual fear conditioning and social interaction memory. For contextual fear conditioning, short- and long-term freezing responses after an electric shock were studied, as well as extinction retention. Regarding social interaction memory, the recognition of a juvenile, a very sensitive short-term memory test, was used. Dopamine loss was carried out by injection of 6-hydroxydopamine, and postmortem catecholamine levels were analyzed by high-performance liquid chromatography. Prefrontocortical dopamine loss (>76%) led to a reactive enhancement of accumbal dopamine content (p<0.01), supporting the hypothesis that a hyperdopaminergic tone emerges in the nucleus accumbens after prefrontocortical dopamine loss. In lesioned rats, long-term extinction of contextual fear conditioning was significantly delayed and extinction retention was impaired without changes in acquisition and short-term contextual fear conditioning and, on the other hand, acquisition and short-term social interaction memory were not affected, although time spent on social interaction was significantly reduced. Added dopamine loss in the nucleus accumbens (>76%) did not alter these behavioral changes. In summary, the results of the present study indicate that the dopaminergic network in the mPFC (but not in the nucleus accumbens) coordinates the normal long-term extinction of contextual fear conditioning responses without affecting their acquisition, and it is involved in time spent on social interaction, but not acquisition and short-term social interaction memory. PMID- 12629529 TI - Disruption and potentiation of latent inhibition by risperidone: the latent inhibition model of atypical antipsychotic action. AB - Latent inhibition (LI), that is, retarded conditioning to a stimulus following its nonreinforced pre-exposure, is impaired in some subsets of schizophrenia patients and in amphetamine-treated rats. Potentiation of LI by antipsychotic drugs (APDs) given in conditioning, under conditions that do not lead to LI in controls, is a well-established index of antipsychotic activity. Recently, we have shown that the atypical APD, clozapine, in addition disrupts LI if administered in pre-exposure, under conditions that lead to LI in controls. This study demonstrates the same behavioral profile for the atypical APD risperidone. LI was measured in a thirst-motivated conditioned emotional response procedure by comparing suppression of drinking in response to a tone previously paired with a foot shock in rats that received nonreinforced exposure to the tone prior to conditioning (pre-exposed (PE)) and rats for whom the tone was novel (non-pre exposed (NPE)). We show that under conditions that did not yield LI in vehicle controls (40 pre-exposures and five conditioning trials), risperidone (0.25, 0.5, and 1.2 mg/kg) led to LI when administered in conditioning. Under conditions that led to LI in vehicle controls (40 pre-exposures and two conditioning trials), risperidone (0.25, 0.5, and 2.5 mg/kg) abolished LI when administered in pre exposure; the latter effect was not evident with haloperidol. In addition, the effects of risperidone administered in both the pre-exposure and conditioning stages were dose-dependent so that the pre-exposure-based action was manifested at lower but not at higher doses. It is concluded that atypical APDs exert in the LI model a dual pattern of effects, which enables detection of their 'typical' action (conditioning-based LI potentiation) as well as a dissociation from typical APDs by their 'atypical' action (pre-exposure-based LI disruption). It is suggested that the former and latter effects are subserved by D2 and 5HT2A antagonism, respectively. PMID- 12629530 TI - The GABA(B) receptor agonist baclofen attenuates cocaine- and heroin-seeking behavior by rats. AB - Conditioned stimuli paired with drugs of abuse can acquire motivational properties, and are capable of inducing drug-seeking behavior and relapse to cocaine use. Converging evidence implicates the mesolimbic dopamine (DA) system, through interactions with limbic afferents to the nucleus accumbens, in behavior controlled by conditioned stimuli. The GABA(B) receptor agonist baclofen has been shown to decrease break points in rats responding for cocaine under progressive ratio schedules and also to attenuate activation of limbic cortical areas in human cocaine addicts. The purpose of the present study was therefore to investigate the effects of baclofen on drug-associated cue-controlled cocaine- or heroin-seeking behavior by rats. Under the second-order schedule of reinforcement used in the present study, cocaine or heroin were available after a fixed time interval, while high rates of responding during the interdrug intervals were maintained by the response-contingent presentations of drug-associated conditioned reinforcers. Baclofen decreased stimulus-maintained responding for either heroin or cocaine, but decreased only cocaine intake under an FR1 schedule. These results therefore support preliminary clinical findings and suggest that drugs with GABA(B) receptor agonist properties may aid abstinence in human drug addicts by decreasing the propensity to cue-induced drug-seeking and relapse. PMID- 12629531 TI - H1-histamine receptor affinity predicts short-term weight gain for typical and atypical antipsychotic drugs. AB - As a result of superior efficacy and overall tolerability, atypical antipsychotic drugs have become the treatment of choice for schizophrenia and related disorders, despite their side effects. Weight gain is a common and potentially serious complication of some antipsychotic drug therapy, and may be accompanied by hyperlipidemia, hypertension and hyperglycemia and, in some extreme cases, diabetic ketoacidosis. The molecular mechanism(s) responsible for antipsychotic drug-induced weight gain are unknown, but have been hypothesized to be because of interactions of antipsychotic drugs with several neurotransmitter receptors, including 5-HT(2A) and 5-HT(2C) serotonin receptors, H(1)-histamine receptors, alpha(1)- and alpha(2)-adrenergic receptors, and m3-muscarinic receptors. To determine the receptor(s) likely to be responsible for antipsychotic-drug-induced weight gain, we screened 17 typical and atypical antipsychotic drugs for binding to 12 neurotransmitter receptors. H(1)-histamine receptor affinities for this group of typical and atypical antipsychotic drugs were significantly correlated with weight gain (Spearman rho=-0.72; p<0.01), as were affinities for alpha(1A) adrenergic (rho=-0.54; p<0.05), 5-HT(2C) (rho=-0.49; p<0.05) and 5-HT(6) receptors (rho=-0.54; p<0.05), whereas eight other receptors' affinities were not. A principal components analysis showed that affinities at the H(1), alpha(2A), alpha(2B), 5-HT(2A), 5-HT(2C), and 5-HT(6) receptors were most highly correlated with the first principal component, and affinities for the D(2), 5 HT(1A), and 5-HT(7) receptors were most highly correlated with the second principal component. A discriminant functions analysis showed that affinities for the H(1) and alpha(1A) receptors were most highly correlated with the discriminant function axis. The discriminant function analysis, as well as the affinity for the H(1)-histamine receptor alone, correctly classified 15 of the 17 drugs into two groups; those that induce weight gain and those that do not. Because centrally acting H(1)-histamine receptor antagonists are known to induce weight gain with chronic use, and because H(1)-histamine receptor affinities are positively correlated with weight gain among typical and atypical antipsychotic drugs, it is recommended that the next generation of atypical antipsychotic drugs be screened to avoid H(1)-histamine receptors. PMID- 12629532 TI - Treatment of weight gain with fluoxetine in olanzapine-treated schizophrenic outpatients. AB - Significant weight gain is a side effect associated with olanzapine treatment in some patients. We investigated the efficacy of high-dose fluoxetine as a weight reducing agent for patients who develop early weight gain with olanzapine treatment. Patients that gained >/=3% of their baseline weight in the initial 8 weeks of olanzapine treatment (n=31) were randomized to double-blind treatment with placebo or fluoxetine (60 mg/day). Clinical, weight, and weight-related measures were assessed. Fluoxetine failed to demonstrate weight-reducing effects (fluoxetine group: baseline mean 80.5 kg, SD=19.1, last mean=83.5 kg, SD=19.8; placebo group: baseline mean=77.1 kg, SD=12.1, last mean=78.8 kg, SD=10.6; F=1.3; df=1, 18; p=0.3). There were no differential effects in psychopathology, extrapyramidal side effects or weight-related measures between the placebo and fluoxetine groups. Serotonin reuptake inhibitors are probably not a practical option to counteract weight gain induced by atypical antipsychotics. Atypical induced weight gain may result from mechanisms other than 5HT reuptake blockade. PMID- 12629533 TI - Allelic variants of the functional promoter polymorphism of the human serotonin transporter gene is associated with auditory cortical stimulus processing. AB - The loudness dependence (LD) of the auditory-evoked N1/P2 component has been shown to be related to the central serotonergic neurotransmission. Allelic variants in the promoter region of the 5-hydroxytryptamine transporter (5-HTT) gene were shown to modulate serotonergic activity. It was hypothesized that the three genotypes (l/l, s/l, s/s) differ with respect to LD. Allelic variants of the 5-HTT promoter region and LD at the Cz electrode were determined in 185 healthy subjects prospectively. A significant association was found between LD and genotype (ANOVA: F=4.172, p=0.017). Individuals homozygous for the l allele exhibited a weaker LD compared to heterozygous subjects. The results are consistent with the reported association between 5-HTT genotype and serotonin transport capacity in lymphoblasts, and indicate that auditory stimulus processing is associated with genetic variants of the brain serotonergic system. The LD may serve as endophenotype in human serotonin research. PMID- 12629534 TI - Serotonin-related gene polymorphisms and central nervous system serotonin function. AB - Central nervous system (CNS) serotonergic function affects a wide range of biological and behavioral functions affecting health and disease. Our objective in this study was to determine whether functional polymorphisms of the genes that encode for the serotonin transporter promoter (5HTTLPR) and monoamine oxidase A (MAOA-uVNTR) are associated with CNS serotonin turnover-indexed by cerebrospinal fluid levels of 5-hydroxyindoleacetic acid (5-HIAA)-in a community sample of healthy adults. Subjects were 165 community volunteers without current medical or psychiatric illness, stratified with respect to ethnicity, gender, and socioeconomic status who underwent inpatient evaluation in the General Clinical Research Center of a university medical center. A significant ethnicity x genotype interaction (P=0.008) indicated that, compared to the long/long and long/short genotypes, the 5HTTLPR short/short genotype was associated with higher CSF 5-HIAA levels in African Americans, but with lower levels in Caucasians. A gender x genotype interaction (P=0.04) indicated that 5HTTLPR short/short genotype was associated with higher 5-HIAA levels in women but with lower levels in men. MAOA-uVNTR 3.5 and 4 repeat alleles were associated with higher 5-HIAA (P=0.03) levels in men, but were unrelated to 5-HIAA levels in women. These findings suggest that effects of serotonin-related gene polymorphisms on CNS serotonergic function vary as a function of both ethnicity and gender. Further research will be required to determine the mechanism(s) underlying these differential effects. In the meanwhile, both ethnicity and gender should be taken into account in research evaluating effects of these and related polymorphisms on CNS serotonergic function, as well as the broad range of biological and behavioral functions that are regulated by CNS serotonergic function. PMID- 12629535 TI - Safety, pharmacokinetics, and effects on cognitive function of multiple doses of GTS-21 in healthy, male volunteers. AB - This study was designed to determine the safety, tolerability, pharmacokinetics and effects on cognitive function of GTS-21 in healthy, male volunteers. A total of 18 subjects were randomized to GTS-21 (25, 75 and 150 mg) or placebo administered three times daily (first 4 days, once on Day 5) for three, 5-day sessions. GTS-21 was well tolerated up to doses of 450 mg/day, with no clinically significant safety findings. C(max) and the area under the plasma concentration of GTS-21 and the metabolite 4-OH-GTS-21 increased in a dose-related fashion; although considerable intersubject variability occurred, it decreased with continued dosing. GTS-21 showed statistically significant enhancement of three measures of cognitive function (attention, working memory, episodic secondary memory) compared to placebo. A relationship between exposure to GTS-21 and the magnitude of the cognitive response was apparent, with maximal effect approached for doses between 75 and 150 mg three times a day. These data indicate that GTS 21 may represent a novel treatment for dementia. PMID- 12629536 TI - Frequency of positive studies among fixed and flexible dose antidepressant clinical trials: an analysis of the food and drug administration summary basis of approval reports. AB - The assumption that the design of an antidepressant clinical trial affects the outcome of that trial is based on sparse data. We sought to examine if the dosing schedule, either a fixed dose or a flexible dose type, in an antidepressant clinical trial affects the frequency with which antidepressants show statistical superiority over placebo. Randomized, placebo-controlled clinical trials of nine antidepressants approved by the Food and Drug Administration between 1985 and 2000 were reviewed. These trials comprised 9313 depressed patients who participated in 51 antidepressant clinical trials consisting of 92 treatment arms with eventual approved doses. In the flexible dose trials, 59.6% (34/57) of the antidepressant treatment arms were statistically significant compared to placebo, whereas in the fixed dose trials only 31.4% (11/35) of the antidepressant treatment arms were statistically significant compared to placebo (chi(2)=6.9, df=1, p<0.01). These data suggest that the antidepressant dose schedule may influence trial outcome due in part to a significantly lower magnitude of symptom reduction with placebo in flexible dose trials (F=4.08, df=1, 48, p&<0.05) compared to fixed dose trials. Symptom reduction was similar with antidepressants in the flexible and fixed dose trials. Further, the primary function of finding a dose-response relationship was not found among the fixed dose studies. PMID- 12629537 TI - Healthy sleepers are rare: problems and success rates in establishing a control group for sleep studies. AB - A large and representative control group is paramount for establishing normative values for sleep in middle-aged persons. However, it is very difficult to recruit normal sleepers. From 286 volunteers who responded to an advertisement for healthy sleepers, only 47 finished the study. We excluded 221 after evaluation of their medical sleep history, general health data, and results of physical examination. We further excluded 18 volunteers after nighttime polysomnography. We finally eliminated from the study 37.5% of the volunteers who entered the screening procedure, on grounds of their sleep disturbances. Our study reveals that investigators face major difficulties when trying to establish a control group of healthy sleepers from the middle-aged population. Sufficient interest on the part of potential test persons does in fact exist, but their general health and, especially, previously undiscovered sleep disorders-frequently prohibit their inclusion. International and interdisciplinary cooperation is necessary to establish a sufficiently large control group for both clinical and research purposes. PMID- 12629538 TI - Methadone treatment induces attenuation of cerebrovascular deficits associated with the prolonged abuse of cocaine and heroin. AB - Opiate replacement therapy has been useful in reducing heroin use and in keeping patients in treatment programs. However, neuropsychological and neurophysiological effects of this treatment regimen have not been evaluated systematically. To determine whether methadone treatment reduces the magnitude of cerebral blood flow alternations in polysubstance (heroin and cocaine) abusers, we compared blood flow parameters in control subjects (n=26), polysubstance abusers (n=28) maintained on methadone for 24 weeks, and polysubstance abusers (n=22) who were not seeking treatment. Blood flow velocity was recorded from the anterior and middle cerebral arteries using transcranial Doppler sonography on an outpatient visit. The pulsatility index, a measure of cerebrovascular resistance, was significantly (p&<0.05) increased in both groups of polysubstance abusers compared to control subjects. Increased pulsatility in the two groups of substance abusers suggests constriction of the small cortical arteries. Nevertheless, the methadone-maintained polysubstance abusers had significantly lower pulsatility values than the nontreatment substance-abusing group. These findings suggest that maintenance on methadone might have significant beneficial neurovascular effects on this population of patients. PMID- 12629539 TI - Cerebrospinal fluid corticotropin-releasing factor (CRF) and vasopressin concentrations predict pituitary response in the CRF stimulation test: a multiple regression analysis. AB - There is considerable evidence that stress-related psychiatric disorders, including depression and post-traumatic stress disorder (PTSD), are associated with hypersecretion of corticotropin-releasing factor (CRF) within the central nervous system (CNS). One line of evidence that is consistent with central CRF hypersecretion in these disorders is the blunted adrenocorticotropin hormone (ACTH) response to intravenous CRF administration, likely a consequence, at least in part, of downregulation of anterior pituitary CRF receptors. The present study tests the hypothesis that elevated cerebrospinal fluid (CSF) concentrations of CRF and a secondary ACTH secretagogue, arginine vasopressin (AVP), are associated with diminished adenohypophyseal responses in a standard CRF stimulation test. CSF concentrations of CRF and AVP, and plasma ACTH responses to the administration of 1 microg/kg ovine CRF (oCRF) were measured in healthy adult women with and without current major depression and/or a history of significant childhood abuse. The primary outcome measure was ACTH area under the curve (AUC) in the CRF stimulation test. Multiple linear regression was performed to identify the impact of CSF CRF and AVP concentrations upon the pituitary response to CRF stimulation. The regression model explained 56.5% of the variation in the ACTH response to CRF stimulation. The relationship of CSF concentrations of CRF to ACTH responses to CRF were best described by a third-order function that was inversely correlated over most of the range of studied values. The association of ACTH response with CSF concentration of AVP and the dose of oCRF followed second order kinetics. These findings support the hypothesis that central CRF hypersecretion is associated with a blunted ACTH response to exogenously administered CRF, explaining almost 60% of the variation in the ACTH response to CRF. PMID- 12629540 TI - Influence of sleep deprivation on neuroactive steroids in major depression. AB - There is evidence from preclinical and clinical studies that concentrations of neuroactive steroids are altered in depression and normalize after antidepressant pharmacotherapy. However, no data are available concerning the impact of sleep deprivation on the concentrations of neuroactive steroids. A total of 29 drug free patients (12 men, 17 women) suffering from major depression according to DSM IV criteria were treated with partial sleep deprivation (PSD). Response to PSD was defined as a reduction of at least 30% according to the six-item version of the Hamilton depression scale (6-HAMD). Plasma samples were taken the day before and after PSD (days 0 and 1) and after one night of recovery sleep (day 2) at 8:00 am. The samples were quantified for neuroactive steroids by means of a highly sensitive and specific combined gas chromatography/mass spectrometry analysis. There was no influence of PSD on the concentrations of neuroactive steroids either in PSD responders (n=20) or in nonresponders (n=9). However, nonresponders showed significantly higher concentrations of 3alpha,5alpha tetrahydroprogesterone (3alpha,5alpha-THP), 3alpha,5beta-tetrahydroprogesterone (3alpha,5beta-THP), and dehydroepiandrosterone (DHEA) before or after PSD compared to responders. In contrast to antidepressant drugs, which correct the dysequilibrium of neuroactive steroids in major depression within several weeks, PSD does not affect the concentrations of neuroactive steroids either in responders or in nonresponders. PMID- 12629541 TI - Differentiation between autism and multiple complex developmental disorder in response to psychosocial stress. AB - Multiple Complex Developmental Disorder (MCDD) represents a distinct group within the autistic spectrum based on symptomatology. Unlike autistic children, part of MCDD children develop schizophrenia in adult life. Despite the differences, patients of both disorders are mainly characterized by abnormal reactions to their social environment. At the biological level, we showed in a previous study that MCDD children have a reduced cortisol response to psychosocial stress. Given the fact that autistic children clinically show more social impairments, it was hypothesized that they may have even further decreased cortisol responses to psychosocial stress than MCDD patients. Therefore, 10 autistic children were compared to 10 MCDD children and 12 healthy control children in their response to a psychosocial stressor, consisting of a public speaking task. In order to test whether any impairments in the biological stress response are specific for psychosocial stress, the autistic children were compared with 11 MCDD children and 15 control children in their response to a physical stressor, consisting of 10 min of bicycle exercise. Heart rate and salivary cortisol levels were used as indicators of response to the stress tests. Autistic children showed a relatively elevated cortisol response to psychosocial stress, in contrast to MCDD children who showed a reduced cortisol response. No differences in heart rate or cortisol responses to the physical stress test were found. The specific difference between autistic and MCDD children in their cortisol response to psychosocial stress indicates that the disturbed reactions to the social environment observed in these disorders may have different biological backgrounds. PMID- 12629542 TI - Lower cortisol levels in depressed patients with comorbid post-traumatic stress disorder. AB - Post-traumatic stress disorder (PTSD) is often comorbid with major depressive episodes (MDEs) and both conditions carry a higher rate of suicidal behavior. Hypothalamic-pituitary-adrenal (HPA) axis and serotonin abnormalities are associated with both conditions and suicidal behavior, but their inter-relation is not known. We determined cortisol response to placebo or fenfluramine in MDE, MDE and PTSD (MDE+PTSD), and healthy volunteers (HVs) and examined the relation of cortisol responses to suicidal behavior. A total of 58 medication-free patients with MDE (13 had MDE+PTSD) and 24 HVs were studied. They received placebo on the first day and fenfluramine on the second day. Cortisol levels were drawn before challenge and for 5 h thereafter. The MDE+PTSD group had the lowest plasma cortisol, the MDE group had the highest, and HVs had intermediate levels. There were no group differences in cortisol response to fenfluramine. Suicidal behavior, sex, and childhood history of abuse were not predictors of baseline or postchallenge plasma cortisol. Cortisol levels increased with age. This study finds elevated cortisol levels in MDE and is the first report of lower cortisol levels in MDE+PTSD. The findings underscore the impact of comorbidity of PTSD with MDE and highlight the importance of considering comorbidity in psychobiology. PMID- 12629543 TI - Reduced pyramidal cell somal volume in auditory association cortex of subjects with schizophrenia. AB - Subjects with schizophrenia have decreased gray matter volume of auditory association cortex in structural imaging studies, and exhibit deficits in auditory sensory memory processes subserved by this region. In dorsal prefrontal cortex (dPFC), similar in vivo observations of reduced regional volume and working memory deficits in subjects with schizophrenia have been related to reduced somal volume of deep layer 3 pyramidal cells. We hypothesized that deep layer 3 pyramidal cell somal volume would also be reduced in auditory association cortex (BA42) in schizophrenia. We used the nucleator to estimate the somal volume of pyramidal neurons in deep layer 3 of BA42 in 18 subjects with schizophrenia, each of whom was matched to one normal comparison subject for gender, age, and post-mortem interval. For all subject pairs, somal volume of pyramidal neurons in deep layer 3 of dPFC (BA9) had previously been determined. In BA42, somal volume was reduced by 13.1% in schizophrenic subjects (p=0.03). Reductions in somal volume were not associated with the history of antipsychotic use, alcohol dependence, schizoaffective disorder, or death by suicide. The percent change in somal volume within-subject pairs was highly correlated between BA42 and BA9 (r=0.67, p=0.002). Deep layer 3 pyramidal cell somal volume is reduced in BA42 of subjects with schizophrenia. This reduction may contribute to impairment in auditory function. The correlated reductions of somal volume in BA42 and BA9 suggest that a common factor may affect deep layer 3 pyramidal cells in both regions. PMID- 12629547 TI - A polarity complex of mPar-6 and atypical PKC binds, phosphorylates and regulates mammalian Lgl. AB - The evolutionarily conserved proteins Par-6, atypical protein kinase C (aPKC), Cdc42 and Par-3 associate to regulate cell polarity and asymmetric cell division, but the downstream targets of this complex are largely unknown. Here we identify direct physiological interactions between mammalian aPKC, murine Par-6C (mPar-6C) and Mlgl, the mammalian orthologue of the Drosophila melanogaster tumour suppressor Lethal (2) giant larvae. In cultured cell lines and in mouse brain, aPKC, mPar-6C and Mlgl form a multiprotein complex in which Mlgl is targeted for phosphorylation on conserved serine residues. These phosphorylation sites are important for embryonic fibroblasts to polarize correctly in response to wounding and may regulate the ability of Mlgl to direct protein trafficking. Our data provide a direct physical and regulatory link between proteins of distinct polarity complexes, identify Mlgl as a functional substrate for aPKC in cell polarization and indicate that aPKC is directed to cell polarity substrates through a network of protein-protein interactions. PMID- 12629548 TI - Identification of the ubiquitin-protein ligase that recognizes oxidized IRP2. AB - The ubiquitin system is involved in several basic cellular functions. Ubiquitination is carried out by a cascade of three reactions catalysed by the E1, E2 and E3 enzymes. Among these, the E3 ubiquitin-protein ligases have a pivotal role in determining the specificity of the system by recognizing the target substrates through defined targeting motifs. Although RING finger proteins constitute an important family of E3 ligases, only a few post-transcriptional modifications, including phosphorylation, proline hydroxylation and glycosylation, are known to function as recognition signals for E3. Iron regulatory protein 2 (IRP2), a modulator of iron metabolism, is regulated by iron induced ubiquitination and degradation. Here we show that the RING finger protein HOIL-1 functions as an E3 ligase for oxidized IRP2, suggesting that oxidation is a specific recognition signal for ubiquitination. The oxidation of IRP2 is generated by haem, which binds to IRP2 in iron-rich cells, and by oxygen, indicating that the iron sensing of IRP2 depends on the synthesis and availability of haem. PMID- 12629549 TI - Building a cell cycle oscillator: hysteresis and bistability in the activation of Cdc2. AB - In the early embryonic cell cycle, Cdc2-cyclin B functions like an autonomous oscillator, whose robust biochemical rhythm continues even when DNA replication or mitosis is blocked. At the core of the oscillator is a negative feedback loop; cyclins accumulate and produce active mitotic Cdc2-cyclin B; Cdc2 activates the anaphase-promoting complex (APC); the APC then promotes cyclin degradation and resets Cdc2 to its inactive, interphase state. Cdc2 regulation also involves positive feedback, with active Cdc2-cyclin B stimulating its activator Cdc25 (refs 5-7) and inactivating its inhibitors Wee1 and Myt1 (refs 8-11). Under the correct circumstances, these positive feedback loops could function as a bistable trigger for mitosis, and oscillators with bistable triggers may be particularly relevant to biological applications such as cell cycle regulation. Therefore, we examined whether Cdc2 activation is bistable. We confirm that the response of Cdc2 to non-degradable cyclin B is temporally abrupt and switch-like, as would be expected if Cdc2 activation were bistable. We also show that Cdc2 activation exhibits hysteresis, a property of bistable systems with particular relevance to biochemical oscillators. These findings help establish the basic systems-level logic of the mitotic oscillator. PMID- 12629550 TI - Functional analysis of an archaebacterial voltage-dependent K+ channel. AB - All living organisms use ion channels to regulate the transport of ions across cellular membranes. Certain ion channels are classed as voltage-dependent because they have a voltage-sensing structure that induces their pores to open in response to changes in the cell membrane voltage. Until recently, the voltage dependent K+, Ca2+ and Na+ channels were regarded as a unique development of eukaryotic cells, adapted to accomplish specialized electrical signalling, as exemplified in neurons. Here we present the functional characterization of a voltage-dependent K+ (K(V)) channel from a hyperthermophilic archaebacterium from an oceanic thermal vent. This channel possesses all the functional attributes of classical neuronal K(V) channels. The conservation of function reflects structural conservation in the voltage sensor as revealed by specific, high affinity interactions with tarantula venom toxins, which evolved to inhibit eukaryotic K(V) channels. PMID- 12629551 TI - Free fatty acids regulate insulin secretion from pancreatic beta cells through GPR40. AB - Diabetes, a disease in which carbohydrate and lipid metabolism are regulated improperly by insulin, is a serious worldwide health issue. Insulin is secreted from pancreatic beta cells in response to elevated plasma glucose, with various factors modifying its secretion. Free fatty acids (FFAs) provide an important energy source as nutrients, and they also act as signalling molecules in various cellular processes, including insulin secretion. Although FFAs are thought to promote insulin secretion in an acute phase, this mechanism is not clearly understood. Here we show that a G-protein-coupled receptor, GPR40, which is abundantly expressed in the pancreas, functions as a receptor for long-chain FFAs. Furthermore, we show that long-chain FFAs amplify glucose-stimulated insulin secretion from pancreatic beta cells by activating GPR40. Our results indicate that GPR40 agonists and/or antagonists show potential for the development of new anti-diabetic drugs. PMID- 12629552 TI - The Par complex directs asymmetric cell division by phosphorylating the cytoskeletal protein Lgl. AB - To generate different cell types, some cells can segregate protein determinants into one of their two daughter cells during mitosis. In Drosophila neuroblasts, the Par protein complex localizes apically and directs localization of the cell fate determinants Prospero and Numb and the adaptor proteins Miranda and Pon to the basal cell cortex, to ensure their segregation into the basal daughter cell. The Par protein complex has a conserved function in establishing cell polarity but how it directs proteins to the opposite side is unknown. We show here that a principal function of this complex is to phosphorylate the cytoskeletal protein Lethal (2) giant larvae (Lgl; also known as L(2)gl). Phosphorylation by Drosophila atypical protein kinase C (aPKC), a member of the Par protein complex, releases Lgl from its association with membranes and the actin cytoskeleton. Genetic and biochemical experiments show that Lgl phosphorylation prevents the localization of cell fate determinants to the apical cell cortex. Lgl promotes cortical localization of Miranda, and we propose that phosphorylation of Lgl by aPKC at the apical neuroblast cortex restricts Lgl activity and Miranda localization to the opposite, basal side of the cell. PMID- 12629553 TI - Hedgehog signalling within airway epithelial progenitors and in small-cell lung cancer. AB - Embryonic signalling pathways regulate progenitor cell fates in mammalian epithelial development and cancer. Prompted by the requirement for sonic hedgehog (Shh) signalling in lung development, we investigated a role for this pathway in regeneration and carcinogenesis of airway epithelium. Here we demonstrate extensive activation of the hedgehog (Hh) pathway within the airway epithelium during repair of acute airway injury. This mode of Hh signalling is characterized by the elaboration and reception of the Shh signal within the epithelial compartment, and immediately precedes neuroendocrine differentiation. We reveal a similar pattern of Hh signalling in airway development during normal differentiation of pulmonary neuroendocrine precursor cells, and in a subset of small-cell lung cancer (SCLC), a highly aggressive and frequently lethal human tumour with primitive neuroendocrine features. These tumours maintain their malignant phenotype in vitro and in vivo through ligand-dependent Hh pathway activation. We propose that some types of SCLC might recapitulate a critical, Hh regulated event in airway epithelial differentiation. This requirement for Hh pathway activation identifies a common lethal malignancy that may respond to pharmacological blockade of the Hh signalling pathway. PMID- 12629554 TI - Ethnic-specific cutpoints for obesity vs country-specific guidelines for action. PMID- 12629555 TI - Endogenous cannabinoid system as a modulator of food intake. AB - The ability of Cannabis sativa (marijuana) to increase hunger has been noticed for centuries, although intensive research on its molecular mode of action started only after the characterization of its main psychoactive component Delta(9)-tetrahydrocannabinol in the late 1960s. Despite the public concern related to the abuse of marijuana and its derivatives, scientific studies have pointed to the therapeutic potentials of cannabinoid compounds and have highlighted their ability to stimulate appetite, especially for sweet and palatable food. Later, the discovery of specific receptors and their endogenous ligands (endocannabinoids) suggested the existence of an endogenous cannabinoid system, providing a physiological basis for biological effects induced by marijuana and other cannabinoids. Epidemiological reports describing the appetite stimulating properties of cannabinoids and the recent insights into the molecular mechanisms underlying cannabinoid action have proposed a central role of the cannabinoid system in obesity. The aim of this review is to provide an extensive overview on the role of this neuromodulatory system in feeding behavior by summarizing the most relevant data obtained from human and animal studies and by elucidating the interactions of the cannabinoid system with the most important neuronal networks and metabolic pathways involved in the control of food intake. Finally, a critical evaluation of future potential therapeutical applications of cannabinoid antagonists in the therapy of obesity and eating disorders will be discussed. PMID- 12629556 TI - Adipose tissue cellularity and apoptosis after intracerebroventricular injections of leptin and 21 days of recovery in rats. AB - OBJECTIVE: To determine the effect of leptin and post-treatment recovery on adipose tissue cellularity and apoptosis. In addition, to investigate whether Bcl 2 and/or Bax is involved in the mechanism of leptin-induced adipose tissue apoptosis. DESIGN: A total of 24 adult male Sprague-Dawley rats were injected i.c.v. with either 10 microg mouse leptin or 10 microl vehicle once per day for 4 days. At 24 h after the last injection, one group was killed while the other was monitored for 21 days. MEASUREMENTS: DNA fragmentation and Bcl-2 and Bax protein levels were determined in inguinal (ING), epididymal (EPI) and retroperitoneal (RP) white adipose tissues and the interscapular brown adipose tissue (BAT). Cellularity was determined in ING and EPI. RESULTS: Leptin significantly reduced the masses of all white fat pads [RPINGEPI] but not BAT. Cell volume was significantly reduced in EPI and ING. Only ING had a significantly reduced cell number from leptin treatment plus exhibited apoptosis by increased DNA fragmentation and DNA laddering, and upregulation of pro-apoptosis Bax protein. The other fat pads exhibited a general trend to increase the Bcl-2/Bax ratio. Recovery allowed for normalization of white fat pad mass, cell number and cell volume; however, BAT mass increased 42% over control. After recovery, apoptosis was not detected, Bcl-2 protein had increased in ING, and the Bcl-2/Bax ratio had risen overall. CONCLUSIONS: Central administration of mouse leptin in the rat targets white fat depots individually to reduce mass by a reduction in cell volume plus adipocyte deletion in, at least, the ING fat pad by Bax-mediated apoptosis. Even after a dramatic loss in adipose tissue mass and change in cellularity, the rat demonstrates a resilient return to control levels together with an increase in factors that prevent adipocyte loss. PMID- 12629557 TI - Entry of exendin-4 into brain is rapid but may be limited at high doses. AB - OBJECTIVE: Peripherally administered exendin-4 is in clinical trials for the treatment of diabetes mellitus and obesity. Since its effects on food intake are mediated centrally, we determined the degree and type of its blood-to-brain penetration of the mouse blood-brain barrier (BBB). MEASUREMENTS AND RESULTS: High-performance liquid chromatography showed that exendin-4 was stable in blood, with most of the injected peptide reaching the brain intact. Capillary depletion studies with washout showed that the injected exendin-4 reached brain parenchyma rather than being trapped in the endothelial cells composing the BBB. Multiple time regression analysis showed that exendin-4 crossed the BBB directly at a fast rate. The rapid brain entry of exendin-4, helped by its high lipophilicity as demonstrated by the octanol/buffer partition coefficient, was not dependent upon circumventricular organs and was not affected by food deprivation for 24 h. The simultaneous i.v. injection of high doses of unlabeled exendin-4 resulted in self inhibition (saturation) that only became statistically significant (P<0.05) when the results of four experiments were combined; this suggests a possible limit to the amount of peripherally administered exendin-4 that can reach the brain after injection of high doses. CONCLUSION: The results indicate that exendin-4 is well conformed for exerting central effects involved in the control of obesity. PMID- 12629558 TI - Inhibitory effect of the pancreatic lipase C-terminal domain on intestinal lipolysis in rats fed a high-fat diet: chronic study. AB - OBJECTIVE: Previous in vitro experiments, as well as acute assays in rat showed that the C-terminal domain (CT-domain) of porcine pancreatic lipase behaves as a potent specific noncovalent inhibitor of pancreatic lipase. Nevertheless, the potential use of the CT-domain as a therapeutic tool against obesity in humans requires further investigation and would be best achieved using the human CT domain. In the present study, we investigated the inhibitory effects of the recombinant human CT-domain, in vivo, upon chronic administration to rats fed a high-fat diet. DESIGN AND MEASUREMENT: The long-term in vivo study requiring relatively high amounts of the human CT-domain, the domain was overexpressed in Escherichia coli as inclusion bodies and an efficient refolding protocol was designed. The inhibitory effect of the recombinant human CT-domain on the activity of pancreatic lipase from different species was first investigated in vitro. Then chronic assays were performed for 4 weeks in rats fed a high-fat diet with or without a daily dose of 1.2 mg of CT-domain per kilogram rat. The time course of food intake, body weight, plasma parameters, liver lipids, faecal output of fat and total cholesterol were measured. RESULTS: A high yield of correctly folded recombinant human CT-domain was obtained using our refolding process, as evidenced by the capability of the recombinant domain to inhibit human horse and porcine pancreatic lipases in vitro. The recombinant human CT domain had no influence on the food intake, but significantly reduced the body weight gain. As compared to control rats, higher amounts of total fat (mainly triglycerides and monoglycerides) and total cholesterol were found in the faeces of the rats treated with the CT-domain. Finally, a decrease in liver triglycerides and nonesterified cholesterol was observed while no significant effect could be detected on the plasma parameters. CONCLUSIONS: These results demonstrated that the CT-domain efficiently reduces in vivo, lipolysis and subsequently body weight gain in rat fed a high-fat diet. The CT-domain could, therefore, be effective in preventing obesity. PMID- 12629559 TI - Strength training for obesity prevention in midlife women. AB - OBJECTIVE: The primary goal of this study was to assess whether increases in fat free mass (FFM) and decreases in total and percentage fat mass from 15 weeks of twice weekly supervised strength training would be maintained over 6 months of unsupervised exercise in a randomized controlled trial. DESIGN: In all, 60 women aged 30-50 y, body mass index between 20 and 35 kg/m(2), were randomized to control or treatment groups. The treatment group performed twice-weekly supervised strength training followed by 6 months of unsupervised training. Measurements at baseline, 15, and 39 weeks included body weight and body composition by dual-energy X-ray absorptiometry. Repeated measures regression was used to assess between-group differences for changes over time. RESULTS: Almost 90% of prescribed exercise sessions were completed. The body composition treatment effects over 15 weeks were largely maintained over 6 months of unsupervised exercise. Over the total 39 weeks of strength training, the treatment group gained +0.89 kg more in FFM, lost -0.98 kg more in fat mass, and lost -1.63% more in percent body fat when compared to the control group. P-values for these between-group differences in 39-week changes were 0.009, 0.06, and 0.006, respectively. Strength training did not result in any significant weight loss or waist circumference attenuation. Adjustment for changes in energy intake and physical activity did not alter these results. CONCLUSIONS: Twice-weekly strength training is behaviorally feasible for busy midlife women and the favorable body composition changes resulting from supervised strength training can be maintained over time. These findings lay the groundwork for determining the long-term health benefits of this behaviorally feasible exercise prescription, potentially including prevention of age-associated fat gains. PMID- 12629560 TI - Genotype-by-smoking interaction for leptin levels in the Metabolic Risk Complications of Obesity Genes project. AB - RATIONALE: Recently, we identified a genotype-by-smoking status interaction with serum leptin levels in a sample of Mexican Americans. However, it is unknown whether this phenomenon occurs in other populations as well. OBJECTIVE: The goal of this study was to examine the genetic architecture of the response to smoking in leptin levels using data from Midwestern Caucasian subjects participating in the Metabolic Risk Complications of Obesity Genes project. METHODS: We employed a variance decomposition analysis using maximum likelihood methods to model genotype-by-smoking interactions for leptin levels and examined the impact of the exclusion of smokers in a subsequent linkage analysis. RESULTS: We found significant evidence (p-value=0.027) for a genotype-by-smoking status interaction for serum leptin levels. In the subsequent linkage analysis with smokers excluded, we obtained a maximum LOD score of 3.4 (P=0.00004) near D8S1128. CONCLUSIONS: These results suggest that a QTL on chromosome 8 may have a differential effect on the expression of leptin in smokers vs nonsmokers, as first identified in Mexican Americans. PMID- 12629561 TI - Which method should be used to determine the obesity, in patients with coronary artery disease? (body mass index, waist circumference or waist-hip ratio). AB - OBJECTIVE: To determine body mass index (BMI), waist circumference and waist-hip ratio (WHR) in cases with angiographically established coronary artery disease (CAD) and to compare the obesity degrees established according to the ranges determined by the International Guidelines Committees for BMI, waist circumference and WHR. DESIGN: Cross-sectional, observational study. SUBJECTS: A total of 617 consecutive cases (516 males, mean age: 57.2+/-10.8 y) with CAD who underwent their first coronary angiography between January 2000 and May 2000. MEASUREMENTS: Before coronary angiography, their heights, weights, waist and hip circumferences were measured. Waist circumferences, BMI and WHRs were compared both as a whole and also within stratified groups as for sexes and age groups categorized in decades above 40 y of age. RESULTS: Overweight cases comprised approximately half of the patients in both sexes. In males, the percentages of obese cases with respect to BMIs were 15%, while males with action level 2 waist circumferences were detected to be 20%. Obese male patients whose WHRs were >or=0.95 were found to be 51%. In female cases, corresponding percentages of obesity were estimated to be 32, 72 and 86%, respectively. CONCLUSION: In the same patient groups, the prevalence of obesity, defined by BMI, waist circumference and WHR, could vary three-fold. Thus, some patients may not display a diagnosis of obesity. To prevent this possibility the waist circumference is an easy method, which reflects central obesity more accurately. PMID- 12629562 TI - Relation of waist circumference and glycemic status to C-reactive protein in the Sandy Lake Oji-Cree. AB - OBJECTIVE: To determine the prevalence of elevated C-reactive protein (CRP) in the Sandy Lake Oji-Cree, an aboriginal community residing in the Sioux Lookout zone of Northwestern Ontario, Canada, and to determine the associations of obesity and diabetes with CRP in a community with a very high prevalence of type II diabetes. DESIGN: We surveyed 512 community members aged 18 y and older to determine the prevalence and the determinants of elevated CRP in Sandy Lake. MEASUREMENTS: Clinical variables, indices of obesity and serum concentrations of CRP, insulin, serum amyloid A (SAA) and interleukin-6 (IL-6). RESULTS: The prevalence of CRP >or=3.8 mg/l was significantly higher in women than in men (51% vs. 32%, P<0.0001). Regression analysis determined that body mass index and IL-6 were independent determinants of CRP concentration in women and waist circumference and IL-6 were independent determinants of CRP concentration in men. Diabetes was associated with elevated CRP in both sexes, but was only a moderate strong determinant in CRP concentration in multivariate regression analysis. CONCLUSIONS: The prevalence of elevated CRP in this aboriginal community is remarkably high. These data further demonstrate that the association between CRP and specific indices of obesity and metabolism vary according to gender and glycemic status. PMID- 12629563 TI - Acute-phase response and immunological markers in morbid obese patients and patients following adjustable gastric banding. AB - OBJECTIVE: We measured markers of acute-phase response and immunological markers in morbid obese patients and in formerly morbid obese patients after a massive weight loss following adjustable gastric banding (GB). SUBJECTS: A total of 49 morbid obese female patients with a body mass index (BMI) above 40 kg/m(2) were investigated during a study period of 6 months. Of these, 24 patients received a gastric banding (GB) and lost a minimum of 20 kg in 1 y (GB group) and 25 patients maintained their weight (obese group). In sum, 13 normal weight subjects (BMI<24 kg/m(2)) were taken for controls. METHOD: Plasma concentration of the acute-phase proteins, C-reactive protein (CRP), orosomucoid, complement factors C3 and C4 and white blood cell count, lymphocyte subsets and serum immunoglobulins were analyzed. RESULTS: Acute-phase proteins were significantly lower in GB compared to morbid obese patients and remained significantly elevated in GB compared to controls. In addition, leukocytes, polymorphonuclear leukocytes and lymphocytes were significantly lower after GB and reached levels comparable to controls (except PMN). No difference in CD3 counts was observed in the three groups. CD4 increased and CD8 decreased in obese and GB patients when compared to controls whereas no statistical difference was found between obese and GB patients. CONCLUSION: Our results confirm the positive effect of GB followed by a massive weight loss without apparent malnutrition. Subclinical chronical inflammation in morbid obese patients leads to irregularities in leukocyte and lymphocyte subsets. These alterations can be positively influenced by GB. PMID- 12629564 TI - Relation of parental history of coronary heart disease to obesity in young adults. AB - OBJECTIVE: To assess the association between obesity and parental coronary heart disease (CHD) history. DESIGN: Analysis of data from an ongoing, large-scale survey on medical status, health behaviour and attitudes. SUBJECTS: Representative samples of Israeli military personnel upon discharge from compulsory service at age 20-22 y. Overall 14297 men and 11638 women were interviewed and examined upon release from military service between 1989 and 1999. MEASUREMENTS: Data on demographic characteristics, family history of CHD, lifestyle, weight and height were collected. Analysis of variance and logistic regression were used. RESULTS: Higher mean body mass index (BMI) and obesity (BMI>30 kg/m(2)) were associated with paternal CHD history in both sexes, and with maternal CHD history among men. Offspring of a parent with a positive CHD history had a higher mean BMI (23.22 vs 22.86 kg/m(2), P<0.001) and were more likely to be obese (5.4 vs 3.7%, P<0.001) than offspring of parents with no history of CHD. Multivariate adjustment for demographic and behavioural variables associated with obesity attenuated the association (adjusted odds ratio for obesity 1.37, 95% confidence interval: 1.15, 1.64). When stratified by sex, this association remained statistically significant only among males. CONCLUSION: Young adults with a parental history of CHD are more likely to be overweight. This high-risk group should be targeted for early preventive activities. PMID- 12629565 TI - The impact of weight change on cardiovascular disease risk factors in young black and white adults: the CARDIA study. AB - OBJECTIVE: To quantify the relation between weight change and change in blood pressure, lipids and insulin levels, and determine if this relation differs by race or initial level of obesity. DESIGN: Longitudinal cohort study. SETTING AND PARTICIPANTS: Community-based sample of 3325 black and white men and women aged 18-30 y from four centers followed for 10 y. Women pregnant at baseline or 10th year exam and persons without a recorded weight at both exams were excluded. Participants whose baseline BMI was >or=25 kg/m(2) were classified as overweight. Height, weight, HDL-cholesterol (HDL-C), LDL-cholesterol (LDL-C), fasting triglycerides, fasting insulin, and blood pressure were measured at baseline and Year 10. RESULTS: The mean (s.d.) of weight gained over 10 y was 10.5 (10.0) kg (black men), 11.7 (11.0) (black women), 7.7 (8.0) (white men), and 7.2 (10.0) (white women). An increase in weight was associated with adverse changes in all factors in all race-sex groups. For example, a 9.1 kg (20-lb) weight increase in persons not overweight at baseline predicted an increase in LDL-C ranging from 0.23 mmol/l in black women to 0.28 mmol/l in black men and a decrease in HDL-C from 0.09 mmol/l (white women) to 0.11 mmol/l (white men) (all P<0.0001). The estimated change in triglycerides was greater in white than in black participants (P<0.02); no other racial differences were found. Changes in triglycerides (P<0.00001) and fasting insulin (P=0.004) were greater in men than in women. Only for LDL-C was a weight change-associated increase significantly different (greater, P<0.001) for nonoverweight persons than for those overweight at baseline. None of these associations were highly specific. Mean levels of LDL-C, HDL-C, and systolic blood pressure improved among all those who lost or did not gain weight. CONCLUSIONS: A 10 y weight gain in young adults of both races and sexes tends to confer adverse changes in their levels of LDL-C, HDL-C, triglycerides, fasting insulin, and blood pressure. This effect occurs regardless of initial weight, age, race, or gender. PMID- 12629566 TI - Study on Lifestyle Intervention and Impaired Glucose Tolerance Maastricht (SLIM): preliminary results after one year. AB - AIMS: Important risk factors for the progression from impaired glucose tolerance to type II diabetes mellitus are obesity, diet and physical inactivity. The aim of this study is to evaluate the effect of a lifestyle-intervention programme on glucose tolerance in Dutch subjects with impaired glucose tolerance (IGT). METHODS: A total of 102 subjects were studied, randomised into two groups. Subjects in the intervention group received regular dietary advice, and were stimulated to lose weight and to increase their physical activity. The control group received only brief information about the beneficial effects of a healthy diet and increased physical activity. Before and after the first year, glucose tolerance was measured and several other measurements were done. RESULTS: Body weight loss after 1 y was higher in the intervention group. The 2-h blood glucose concentration decreased 0.8+/-0.3 mmol/l in the intervention group and increased 0.2+/-0.3 mmol/l in the control group (P<0.05). Body weight loss and increased physical fitness were the most important determinants of improved glucose tolerance and insulin sensitivity. CONCLUSION: A lifestyle-intervention programme according to general recommendations is effective and induces beneficial changes in lifestyle, which improve glucose tolerance in subjects with IGT. Body weight loss and increased physical fitness were the most important determinants of improved glucose tolerance and insulin sensitivity. PMID- 12629567 TI - A novel nonsense mutation in the melanocortin-4 receptor associated with obesity in a Spanish population. AB - BACKGROUND: In recent years, several groups have reported dominant inheritance of obesity conferred by missense, nonsense and frameshift mutations in the melanocortin 4 receptor gene (MC4R). Hence, MC4R is involved in the most common monogenic form of human obesity described so far. OBJECTIVES: In this context, we screened a Spanish population, composed of obese subjects and normal weight controls, for mutations in the MC4-R by single-strand conformational polymorphism (SSCP). SUBJECTS AND METHODS: Overall 313 individuals, 159 obese subjects (body mass index: BMI: 37.6 kg/m(2), 95% CI: 36.7-38.5 kg/m(2)) and 154 normal weight control subjects (BMI: 22.3 kg/m(2), 95% CI: 22.0-22.6 kg/m(2)) were screened for MC4-R mutations. RESULTS: We detected a novel nonsense mutation at codon 16 of the MC4-R in an obese female (BMI: 30.0 kg/m(2)) and a previously described missense mutation (Val-253-Ile) located within the sixth trans-membrane domain of the MC4-R in a normal weight individual (BMI: 19.0 kg/m(2)). The polymorphism Val 103-Ile was detected in one obese individual, while four subjects (two cases and two controls) with the polymorphism Ile-251-Leu were found. CONCLUSIONS: We have identified a novel nonsense mutation (Trp-16-Stop) that, based on previously described frameshift and nonsense mutations, most likely results in dominantly inherited obesity. Within this Spanish population, the frequency of the Ile-251 Leu polymorphism of the MC4R was similar in obese and control subjects (about 1.3%), while the polymorphism Val-103-Ile was only detected in an obese individual (0.6%). PMID- 12629568 TI - Interactions among the beta2- and beta3- adrenergic receptor genes and total body fat and abdominal fat level in the HERITAGE Family Study. AB - OBJECTIVE AND SUBJECTS: Interactions between markers in the beta2- and beta3 adrenergic receptor (ADR) genes and total body fat and computerized tomography measured abdominal fat phenotypes were studied in the HERITAGE Family Study cohort of Black (n=205; 81 males and 124 females) and White (n=415; 198 males and 217 females) subjects before and after an endurance training program. RESULTS: In Black subjects, beta2- and beta3-ADR gene variants showed evidence of interactions on changes in total body fat mass and abdominal fat area (P<0.005 and =0.010, respectively). Black subjects who were carriers of both beta2-ADR Arg16 and beta3-ADR Arg64 alleles had a greater decrease in total fat mass as well as abdominal total and subcutaneous, but not visceral fat areas in response to endurance training than subjects with other genotype combinations (P from 0.011 to 0.047). After correction for multiple tests, the findings remained essentially unchanged for total body fat mass and abdominal fat area, but became nonsignificant for subcutaneous fat area. The changes in abdominal fat correlated positively with the changes in fat mass (P<0.0001). The interactions between beta2 and beta3-ADR gene markers accounted for a maximum of 3% of the variances in the response of total fat mass and abdominal fat area to endurance training in Black subjects but it was not significant in White subjects. CONCLUSION: Interactions between sequence variants in the beta2-beta3-ADR gene contributed to the changes in fat mass and abdominal adiposity in response to endurance training in Black subjects. PMID- 12629569 TI - Social factors and obesity: an investigation of the role of health behaviours. AB - OBJECTIVES: This study evaluated a behavioural model of the relation between social factors and obesity, in which differences in body mass index (BMI) across sociodemographic groups were hypothesized to be attributable to social group differences in health behaviours affecting energy expenditure (physical activity, diet and alcohol consumption and weight control). METHODS: A total of 8667 adults who participated in the 1995 Australian National Health and Nutrition Surveys provided data on a range of health factors including objectively measured height and weight, health behaviours, and social factors including family status, employment status, housing situation and migration status. RESULTS: Social factors remained significant predictors of BMI after controlling for all health behaviours. Neither social factors alone, nor health behaviours alone, adequately explained the variance in BMI. Gender-specific interactions were found between social factors and individual health behaviours. CONCLUSIONS: These results suggest that social factors moderate the relation between BMI and weight-related behaviours, and that the mechanisms underlying sociodemographic group differences in obesity may vary among men and women. Additional factors are likely to act in conjunction with current health behaviours to explain variation in obesity prevalence across sociodemographic groups. PMID- 12629570 TI - Prevalence of binge-eating disorder in obese children and adolescents seeking weight-loss treatment. AB - OBJECTIVE: The aim of the present study was to examine the extent to which a population of obese children and adolescents developed binge-eating disorder (BED). METHOD: A sample of 196 obese children and adolescents (aged 10-16 y) seeking weight-loss treatment at two treatment facilities (inpatient and outpatient treatment) was screened using the eating disorder examination. RESULTS: : Only 1% of the subjects met the criteria for BED and 9% were found to have objective bulimic episodes (OBEs, overeating with loss of control), but did not endorse all of the other DSM-criteria that are required for a diagnosis of BED. OBEs were more common in girls than in boys. Episodic overeating was more common than binge eating. Compared to children without OBEs, children engaging in OBEs were more overweight and showed a greater eating-related psychopathology. The age of the first OBE was 10.88 y (s.d.=2.60). It appears that overweight precedes binge eating. DISCUSSION: A subgroup of girls and boys seeking treatment for obesity shows considerable eating difficulties. The results highlight the importance of considering binge-eating symptoms when devising treatment programmes for children and adolescents suffering from obesity. PMID- 12629571 TI - Body composition of 4- and 5-year-old New Zealand girls: a DXA study of initial adiposity and subsequent 4-year fat change. AB - BACKGROUND: Dual-energy X-ray information (DXA) quantitating body fat mass and percentage fat in healthy children of preschool age is scarce. OBJECTIVE: To study the initial variability in body composition and subsequent longitudinal changes in absolute fat mass (kg) and relative adiposity (fat percentage) in a sample of contemporary young New Zealand girls. DESIGN: Cross-sectional study with a longitudinal component. SETTING: University research unit. SUBJECTS: A total of 89 Caucasian girls aged 4-5 y were recruited by advertisement at baseline and 4-y changes in body composition were evaluated in 23 of these girls. METHODS: Total body composition was measured by DXA, height and weight by anthropometry. RESULTS: Baseline values for fat mass varied more than values for lean mass or bone mass. Girls from the upper third of our fat percentage distribution (% fat >19.2%) had more than twice the fat mass (5.34 vs 2.31 kg, P<0.001) of those from the lowest third (% fat &<15.4%). The percentage gain in fat mass over 4 y (124 (95% CI 90-163) also exceeded the percentage gain of lean mass (55 (95% CI 51-59). In data adjusted for age and height, 63.5% of the variance in percentage body fat at time 2 was explained by fat mass at time one. CONCLUSIONS: In girls, the trajectory of fat gain appears to be established at a young age. Our results support the view that body fatness tracks strongly before puberty. Since preventing the accumulation of excessive fat is preferable to reduction of existing excessive fat stores, it is important to put in place strategies to limit excessive fat gain early in life. PMID- 12629572 TI - Greater prevalence of iron deficiency in overweight and obese children and adolescents. AB - OBJECTIVE: To assess whether overweight children and adolescents, who often have poor dietary habits, are at increased risk of iron deficiency (ID). METHODS: The study sample included 321 children and adolescents followed in two endocrine centers in Israel between 1999 and 2001. The subjects were divided into three groups on the basis of body mass index (BMI) for age and gender as follows: group 1-BMI below 85th percentile (normal weight); group 2-BMI above 85th, but below 97th percentile (overweight); and group 3-BMI above 97th percentile (obese). ID was defined as iron levels <8 micromol/l (45 mcg/dl), and iron-deficiency anemia (IDA) was defined as ID and hemoglobin level below 2 standard deviation score (SDS) for the mean for age and gender. RESULTS: Iron levels below 8 micromol/l (45 mcg/dl) were noted in 38.8% of the obese children and 12.1% of the overweight children, compared with 4.4% of the normal-weight group (P<0.001). There was a significant negative correlation of low iron levels with BMI SDS (r=-0.44, P<0.001), but not with age or gender. Among the children with ID, 26.6% also had IDA. Groups 1, 2, and 3 accounted for 6.7%, 35%, and 58.3% of the children with IDA, respectively. CONCLUSIONS: ID is common in overweight and obese children. A significantly greater proportion of obese than normal-weight children have IDA. Insufficient dietary intake of iron, whether absolute or relative to body mass, and increased iron needs may be a result of unbalanced nutrition or repeated short-term restrictive diets. Because of potentially harmful effects of ID, obese children should be routinely screened and treated as necessary. PMID- 12629575 TI - Cytochrome P450 3A pharmacogenetics: the road that needs traveled. PMID- 12629576 TI - Polymorphic CYP2A6 and its clinical and toxicological significance. PMID- 12629577 TI - The challenges of recording phenotype in a generalizable and computable form. PMID- 12629578 TI - Attention-deficit/hyperactivity disorder: current aspects on pharmacogenetics. PMID- 12629579 TI - Pharmacogenetics: the ethical issues. PMID- 12629580 TI - Sequence variations in the UDP-glucuronosyltransferase 2B7 (UGT2B7) gene: identification of 10 novel single nucleotide polymorphisms (SNPs) and analysis of their relevance to morphine glucuronidation in cancer patients. AB - We have screened a cohort of 239 Norwegian cancer patients for sequence variation in the coding and regulatory regions of the UDP-glucuronosyltransferase 2B7 gene (UGT2B7) and analyzed the impact of gene variants on morphine glucuronidation in vivo. In all, 12 single nucleotide polymorphisms (SNPs) were identified, 10 of which have not been previously described. Only one SNP causes a change in amino acid sequence (H268Y). Seven UGT2B7 genotypes were observed and three main haplotypes predicted. There was no correlation between UGT2B7 genotype or haplotype and morphine glucuronide to morphine serum ratios among 175 patients who received chronic oral morphine therapy, and who had normal renal and hepatic function. The apparent lack of functional polymorphisms fits well with the near unimodal, but broad, distributions of the ratios (morphine 3 glucuronide/morphine: 6.4-309.2; morphine 6-glucuronide/morphine: 0.5-72.8). Our results suggest that factors other than UGT2B7 polymorphism may be more deciding for the variability in morphine glucuronide to morphine serum ratios. PMID- 12629582 TI - Analysis of gene expression in carbon tetrachloride-treated rat livers using a novel bioarray technology. AB - The present study successfully utilizes a new ADME Rat Expression Bioarray, containing 1040 metabolism- and toxicology-linked genes, to monitor gene expression from the livers of rats treated with carbon tetrachloride (CCl(4)). Histopathological analysis, hierarchical clustering methods, and gene expression profiling are compared between the control and CCl(4)-treated animals. A total of 44 transcripts were found to be altered in response to the hepatotoxin, 19 of which were upregulated and 25 were downregulated. Some of these gene expression changes were expected and concurred with previously published data while others were novel findings. PMID- 12629581 TI - Transcriptional profiling in the human prefrontal cortex: evidence for two activational states associated with cocaine abuse. AB - CNS-focused cDNA microarrays were used to examine gene expression profiles in dorsolateral prefrontal cortex (dlPFC, Area 46) from seven individual sets of age and post-mortem interval-matched male cocaine abusers and controls. The presence of cocaine and related metabolites was confirmed by gas chromatography-mass spectrometry. Sixty-five transcripts were differentially expressed, indicating alterations in energy metabolism, mitochondria and oligodendrocyte function, cytoskeleton and related signaling, and neuronal plasticity. There was evidence for two distinct states of transcriptional regulation, with increases in gene expression predominating in subjects testing positive for a metabolite indicative of recent 'crack' cocaine abuse and decreased expression profiles in the remaining cocaine subjects. This pattern was confirmed by quantitative polymerase chain reaction for select transcripts. These data suggest that cocaine abuse targets a distinct subset of genes in the dlPFC, resulting in either a state of acute activation in which increased gene expression predominates, or a relatively destimulated, refractory phase. PMID- 12629583 TI - Role of polymorphic human CYP2B6 in cyclophosphamide bioactivation. AB - The role of polymorphic CYP2B6 in cyclophosphamide (CPA) bioactivation was investigated in human liver microsomes. A total of 67 human liver specimens were first genotyped with respect to the CYP2B6*5 and CYP2B6*6 variant alleles. CYP2B6 apoprotein levels in 55 liver microsomal preparations were assessed by immunoblotting. 4-Hydroxy-CPA and hydroxy-bupropion were quantified by using HPLC and LC-MS, respectively. 7-Ethoxy-4-trifluoromethyl coumarin O-deethylase activity was measured fluorometrically. The frequencies of CYP2B6*5 and CYP2B6*6 mutant alleles were 9.0 and 16.4%, respectively. CYP2B6 protein expression was detected in 80% of the samples, with a large variation (0.003-2.234, arbitrary units). There was a high correlation between CYP2B6 apoprotein content and CPA 4 hydroxylation (n=55, r=0.81, P<0.0001). When based on the CYP2B6 apoprotein levels, the *6 carriers had significantly higher CPA 4-hydroxylation (P<0.05). CPA 4-hydroxylation also correlated significantly with other CYP2B6-specific reactions (n=20, P<0.0001). V(max) and K(m) for CPA 4-hydroxylation in recombinant CYP2B6 enzyme were 338 nmol/min/nmol enzyme and 1.4 mM, respectively. CYP2B6 showed much higher in vitro intrinsic clearance than previously observed in recombinant CYP2C19 and CYP2C9 variants in yeast expression system. Our results demonstrate that the polymorphic CYP2B6 is a major enzyme in the bioactivation of CPA. Moreover, we identified a strong impact of CYP2B6*6 on CPA 4-hydroxylation. PMID- 12629584 TI - Hypertension therapeutics in turmoil. PMID- 12629585 TI - Resolution of ST-segment elevation after thrombolytic therapy in elderly patients with acute myocardial infarction. AB - We examined the resolution of ST-segment elevation after thrombolytic therapy in elderly versus younger patients with acute myocardial infarction. Electrocardiograms were recorded before, on completion of, and on day 1 and day 2 post-thrombolytic therapy (streptokinase or tissue thromboplastin activator) in 36 patients older than 65 years and 36 patients younger than 65 years. There was no significant different in the pre-thrombolytic ST-segment elevation per lead in both elderly and younger patients (3.7 +/- 0.7 versus 3.5 +/- 0.8 mm; P = NS). On completion of thrombolytic therapy, both groups demonstrated resolution of ST segment elevation and, although the ST-segment elevation per lead was higher in elderly patients (3.0 +/- 0.9 versus 2.5 +/- 0.9 mm; P = 0.008), the percentage resolution per lead was not significantly different (19% versus 29%; P = NS). On day 1 post-thrombolytic therapy, there was further resolution of ST-segment elevation in both groups, but at this point, the percentage resolution per lead was significantly less in the elderly than in the younger patients (51% versus 66%; P = 0.03), and the ST-segment elevation per lead remained higher in elderly patients (1.8 +/- 1.0 versus 1.2 +/- 0.6 mm; P = 0.0009). On day 2 post thrombolytic therapy, although there was further resolution of ST-segment elevation in both groups, the percentage resolution per lead remained significantly less (68% versus 80%; P = 0.05) and ST-segment elevation per lead remained significantly higher in elderly patients (1.2 +/- 0.7 versus 0.7 +/- 0.4 mm; P = 0.0002). Resolution of ST-segment elevation after thrombolytic therapy was less marked in elderly patients, indicating a reduced response to thrombolytic therapy in this patient population. PMID- 12629586 TI - A Bayesian approach for the estimation of pharmacokinetic parameters in children. AB - A Bayesian method is proposed for the first-time assessment of pharmacokinetic parameters (area under the curve [AUC], maximum concentration [C(max)], half-life [T(1/2)]) in children after a single oral dose. Pharmacokinetic parameters were compared using sparse sampling (three blood samples) with extensive sampling (nine blood samples). Two different therapeutic classes of drugs were evaluated in this study. The population pharmacokinetic parameters and the blood sampling time points were obtained from adult data. The results indicate that the Bayesian approach can be used to estimate pharmacokinetic parameters in children with fair degree of accuracy. The Bayesian method described here can be used to assess pharmacokinetic parameters in children, provided there is a prior knowledge of the pharmacokinetics of a drug in adult population. PMID- 12629587 TI - Provision of drug information to patients by pharmacists: the impact of the Omnibus Budget Reconciliation Act of 1990 a decade later. AB - Drug-related illness in the United States factors substantially in health care costs, although often these illnesses and their attendant costs are preventable. One strategy for minimizing adverse drug reactions is to provide drug information to consumers in the form of prescription counseling at pharmacies. The Omnibus Budget Reconciliation Act of 1990 (OBRA 1990) contained provisions for mandating such counseling to Medicaid patients. OBRA 1990 was implemented in 1993, but most states acted quickly to extend counseling services to all patients receiving prescription drugs. We looked at the extent and quality of prescription counseling available in community pharmacies 1 decade after OBRA 1990 was written. We evaluated the counseling services afforded at large chain pharmacies, independent community pharmacies, and on-line pharmacies for a hydrochlorothiazide prescription. We found that most (69%) pharmacies offered to provide prescription counseling service, and that average counseling index scores, a measure of the quality or extent of information provided as determined by a Rasch analysis, were generally satisfactory. Our observations based on a single prescription for hydrochlorothiazide, along with other studies, suggest that there is a positive upward trend in the number of pharmacies providing prescription drug information, and that the extent of information provided suggests that the objectives of OBRA 1990 and related legislation to reduce ADRs are being fundamentally satisfied. PMID- 12629588 TI - Left ventricular diastolic function assessment by tissue Doppler echocardiography in relation to hormonal replacement therapy in postmenopausal women with diastolic dysfunction. AB - The objective of this study was to evaluate the effect of hormone replacement therapy (HRT) regimens on left ventricular diastolic function by using mitral pulsed wave Doppler (MPWD) and tissue Doppler velocities (TDE). Seventy-eight postmenopausal women with normotensive and impaired diastolic left ventricular filling were included in the study. All the patients began a six-cycle HRT course. This formulation consisted of E2 valerate plus Medroxy progesterone acetate (MPA). Left ventricular diastolic function at rest was evaluated by M mode, two-dimensional, MPWD and TDE in 78 postmenopausal women with normal blood pressure before the treatment for 6 months of HRT. The M-mode, two-dimensional, and MPWD parameters assessed were heart rate, systolic blood pressure, diastolic blood pressure, left ventricular mass index, ejection fraction of the left ventricle (EF), septal (IVS) and posterior wall (PW) thickness, left ventricular end-systolic (LVESD) and end-diastolic (LVEDD) diameter, left atrial diameter, peak early diastolic velocity (E), peak atrial velocity (A), E/A ratio, E acceleration time, E deceleration time, diastolic filling period, and isovolumic relaxation time (IVRT). The TDE parameters assessed were peak early diastolic velocity (E'), peak late diastolic velocity (A'), peak systolic velocity, E'/A' ratio, E' acceleration time, E' deceleration time, IVRT', and E/E' ratio. Quantitative data were analyzed using Student t test. Among the MPWD parameters, peak A velocity, E deceleration time, and IVRT significantly decreased, while peak E velocity and E/A ratio increased after a 6-month treatment. From the point of TDE parameters, E' velocity and E'/A' ratio increased, while A' velocity, E' deceleration time, E/E' ratio and IVRT' decreased. Some MPWD and TDE parameters were partially reversed after HRT. TDE velocities and especially E/E' ratio may provide better and true information of the diastolic function. TDE parameters were independent from the preload and did not produce pseudonormal pattern. HRT may cause increase in the blood volume and produce pseudonormal pattern in transmitral flow. In that case, TDE may be a beneficial method for evaluation of diastolic function. PMID- 12629589 TI - Perils of the random experiment. AB - Most medical research is executed on samples selected from large populations. Nevertheless, health care researchers often blur the difference between interpreting sample-based research and evaluating research that included the entire population of interest. This is an implication-critical distinction; in population research, every result applies to the population (because the entire population was included in the analysis), although only a few results from sample based research can be extended to the population at large. Treating every result from sample-based research as if that result applies to the population is misleading. Using nonmathematic terminology, this article develops the reason for the differences in the implications of these two research perspectives. In sample based research, the best indicators of which results should be extended from the sample to the population are the presence of (1) a prospective plan for that experiment; and (2) the execution of the experiment according to that plan (concordant execution). The absence of these two features produces execution and analysis decisions based on the incoming data stream-the hallmark of the random experiment. In this latter paradigm, allowing the data to influence the execution and analysis decisions renders the usual estimates of effect size, standard errors, confidence intervals, and P values untrustworthy. Readers of clinical trial results must be vigilant for nonprotocol-driven research and understand that the results from these programs are at best exploratory and cannot be used to answer scientific questions. PMID- 12629590 TI - Management of refractory hypertension. AB - True refractory hypertension is unusual in clinical practice, thanks to the widespread availability of antihypertensive drugs and national mandate to optimize blood pressure control levels in the community. However, at times the blood pressure may become refractory to initial drug therapy. When the blood pressure levels do not reach a target level despite usual therapy, hypertension is considered refractory. There should be proper evaluation of such patients to determine the factor(s) responsible for resistance to therapy. In many patients, proper adjustment of drug doses, including effective use of diuretics, restores the blood pressure level. For some patients, potent drug therapy, such as hydralazine or minoxidil, must be considered. Based upon clinical course, work-up for secondary causes of hypertension should be considered in selected patients. Refractory hypertension requires proper analysis of etiologic factors and consideration of rational drug selection, and at times, work-up for secondary causes of hypertension. PMID- 12629591 TI - Epidemiology of bacteremia in the geriatric population. PMID- 12629592 TI - Delayed presentation of calcium channel antagonist overdose. AB - The calcium channel antagonists are generally safe in therapeutic dosage, but severe side effects with elevated intake are increasingly described. Typical features include confusion, lethargy, hypotension, sinus node depression, and cardiac conduction defects. Even if patients are stable on presentation, this does not preclude the possible late development of adverse events from the long acting formulations of calcium channel blockers. A case of toxic overdose with 1440 mg of slow-release diltiazem is presented; this patient was stable on presentation, but rapidly became hemodynamically unstable, requiring treatment with intravenous calcium, temporary pacemaker, inotropic support and mechanical ventilation with a successful outcome. A concise review of the therapeutic considerations is provided. PMID- 12629593 TI - Severe bleeding symptoms in refractory idiopathic thrombocytopenic purpura: a case successfully treated with melatonin. AB - The prognosis of patients with idiopathic thrombocytopenic purpura refractory to corticosteroids and splenectomy (refractory ITP) is poor; these patients present high morbidity from disease and its treatment and have a mortality rate of approximately 16%. A patient is reported with severe bleeding symptoms related to refractory ITP successfully treated with melatonin. PMID- 12629594 TI - Radiocontrast-induced nephropathy. AB - Radiocontrast administration remains the third leading cause of hospital-acquired acute renal failure. Clinically, radiocontrast-induced nephropathy (RIN) is defined as a sudden decline in renal function after radiocontrast administration. Typically, the serum creatinine level begins to increase at 24 to 72 hours after the administration of contrast, peaks at 3 to 5 days, and requires another 3 to 5 days to return to baseline. RIN increases the incidence of life-threatening complications such as sepsis, bleeding, and respiratory failure and increases the cost of medical care by extending the hospital stay. The increased mortality associated with acute renal failure encountered in this scenario calls for a heightened awareness of the diagnosis and prevention of RIN. Whereas individuals with healthy renal function are not generally considered to be at particular risk for RIN, patients with preexisting renal insufficiency and diabetes mellitus are much more likely to experience acute renal failure after contrast administration. In the past, a variety of therapeutic interventions have been used to prevent or attenuate RIN, including saline hydration, diuretics, mannitol, calcium channel antagonists, theophylline, endothelin receptor antagonists, hemodialysis, and dopamine. More recently, studies demonstrate a positive impact of fenoldopam (dopamine-1 receptor, dopamine-1 agonist) and the antioxidant N-acetylcysteine in ameliorating RIN. This article discusses the pathophysiology, risk factors, and prevention of RIN. PMID- 12629595 TI - Overview of existing research and information linking isotretinoin (accutane), depression, psychosis, and suicide. AB - Isotretinoin (Accutane; Hoffmann-La Roche, Nutley, NJ) is a drug closely related to the chemical structure of vitamin A. The pharmacology and toxicology of these two retinoids are similar enough to warrant comparison. Accutane is a powerful drug that its manufacturer, Roche, indicates is limited for severe recalcitrant nodular acne. This potency is also reflected in Accutane's well-known ability to produce severe birth defects if taken during pregnancy. Less well known is the risk of this lipid-soluble chemical to affect the central nervous system. Reports of intracranial hypertension, depression, and suicidal ideation with Accutane use have prompted an examination of its serious and life-threatening potential. Although Roche has added a warning to its product label for signs of depression, and suicidal ideation, this product is overprescribed for all forms of acne, including mild and moderate cases that have not been treated with alternative medications with less risk of depression and suicide. There is no contesting that this drug is effective at clearing up the most severe forms of acne, but the public must be informed of the proper limited indication for its use, because depression and suicide can follow in patients with no prior history of psychiatric symptoms or suicide attempts. PMID- 12629597 TI - Deletion of the telomerase reverse transcriptase gene and haploinsufficiency of telomere maintenance in Cri du chat syndrome. AB - Cri du chat syndrome (CdCS) results from loss of the distal portion of chromosome 5p, where the telomerase reverse transcriptase (hTERT) gene is localized (5p15.33). hTERT is the rate-limiting component for telomerase activity that is essential for telomere-length maintenance and sustained cell proliferation. Here, we show that a concomitant deletion of the hTERT allele occurs in all 10 patients with CdCS whom we examined. Induction of hTERT mRNA in proliferating lymphocytes derived from five of seven patients was lower than that in unaffected control individuals (P<.05). The patient lymphocytes exhibited shorter telomeres than age matched unaffected individuals (P<.0001). A reduction in replicative life span and a high rate of chromosome fusions were observed in cultured patient fibroblasts. Reconstitution of telomerase activity by ectopic expression of hTERT extended the telomere length, increased the population doublings, and prevented the end-to-end fusion of chromosomes. We conclude that hTERT is limiting and haploinsufficient for telomere maintenance in humans in vivo. Accordingly, the hTERT deletion may be one genetic element contributing to the phenotypic changes in CdCS. PMID- 12629596 TI - Unraveling monogenic channelopathies and their implications for complex polygenic disease. AB - Ion channels are a large family of >400 related proteins representing >1% of our genetic endowment; however, ion-channel diseases reflect a relatively new category of inborn error. They were first recognized in 1989, with the discovery of cystic fibrosis transmembrane conductance regulator, and rapidly advanced as positional and functional studies converged in the dissection of components of the action potential of excitable tissues. Although it remains true that diseases of excitable tissue still most clearly illustrate this family of disease, ion channel disorders now cover the gamut of medical disciplines, causing significant pathology in virtually every organ system, producing a surprising range of often unanticipated symptoms, and providing valuable targets for pharmacological intervention. Many of the features shared among the monogenic ion-channel diseases provide a general framework for formulating a foundation for considering their intrinsically promising role in polygenic disease. Since an increasingly important approach to the identification of genes underlying polygenic disease is to identify "functional candidates" within a critical region and to test their disease association, it becomes increasingly important to appreciate how these ion-channel mechanisms can be implicated in pathophysiology. PMID- 12629599 TI - Mini-incision technique for total hip arthroplasty with navigation. AB - This prospective study compares a mini-incision technique and traditional posterior approach for total hip arthroplasty (THA). Thirty-three patients who had undergone a mini-incision THA were matched by diagnosis, gender, average age, and preoperative Harris Hip Score (HHS) to 33 patients who had undergone THA using the traditional posterior approach. The average length of the incision for group 1 was 11.7 cm (range, 7.3-13.0) and for group 2 was 20.2 cm (range, 14.8 26.0). At the 3-month follow-up, patients in the mini-incision group had significant improvement in limp (P<.05) and ability to climb stairs (P <.01) compared with the traditional group. At the 6 month follow-up, the mini-incision group was significantly better in terms of limp (P <.05), distance walked (P<.001), and stairs (P < 0.001). There was no significant difference between groups for pain, function, or range of motion at the 1-year follow-up examination. PMID- 12629600 TI - The fate of cementless jumbo cups in revision hip arthroplasty. AB - We report 5-year minimum results of cementless over-sized cups used in revision hip arthroplasty, with significant associated bone defects. Forty-three porous coated jumbo cups were used to treat acetabular defects in revision hip arthroplasty in 42 patients with a mean age of 63 (range, 25-86). Morsellized allograft only was used in 27 hips, and bulk allograft was used in 8 cases. Two patients were lost to follow-up, and 5 died after a mean 7 years' follow-up, with retention of their prostheses. In the remaining 36 cases, the mean follow-up was 10 years (range, 6-14 years). Two acetabular components were revised for aseptic loosening and graft resorption. Two cases were complicated by dislocation. A satisfactory 92% Kaplan Meier shell survival rate was seen at 14 years. PMID- 12629598 TI - Extensive female-mediated gene flow from sub-Saharan Africa into near eastern Arab populations. AB - We have analyzed and compared mitochondrial DNA variation of populations from the Near East and Africa and found a very high frequency of African lineages present in the Yemen Hadramawt: more than a third were of clear sub-Saharan origin. Other Arab populations carried approximately 10% lineages of sub-Saharan origin, whereas non-Arab Near Eastern populations, by contrast, carried few or no such lineages, suggesting that gene flow has been preferentially into Arab populations. Several lines of evidence suggest that most of this gene flow probably occurred within the past approximately 2,500 years. In contrast, there is little evidence for male-mediated gene flow from sub-Saharan Africa in Y chromosome haplotypes in Arab populations, including the Hadramawt. Taken together, these results are consistent with substantial migration from eastern Africa into Arabia, at least in part as a result of the Arab slave trade, and mainly female assimilation into the Arabian population as a result of miscegenation and manumission. PMID- 12629601 TI - Acetabular components without bulk bone graft in revision surgery: A 5- to 13 year follow-up study. AB - Eighty-three consecutive revisions in 74 patients using a porous-coated acetabular component without bulk bone graft were followed up for an average of 9.3 years (range, 5-13 years). A large-diameter cup was implanted in hips with adequate osseous support, and a high hip center technique using a standard- or smaller-diameter cup was selected in hips without sufficient bone stock. Acetabular bone deficiency was segmental in 18 hips, cavitary in 30, and combined segmental and cavitary in 35. Four (5%) cups were revised again; 1 for infection, 1 for dislodgement of the polyethylene liner from the metal shell, and 2 for recurrent dislocation. No acetabular components were categorized as definitely loose at final follow-up. The current satisfactory results encourage the use of this simple technique. PMID- 12629602 TI - A 10-year minimum follow-up of hydroxyapatite-coated threaded cups: clinical, radiographic and survivorship analyses with comparison to the literature. AB - We evaluated the clinical, radiographic, and survivorship outcomes in a series of 418 threaded hydroxyapatite-coated acetabular cups (Arc2f; Osteonics, Allendale, NJ) implanted in a consecutive series of 384 patients undergoing primary total hip arthroplasty. In all cases, the cup was screwed into the prepared acetabulum. Bone screws were used to provide secondary fixation. At a minimum 10-year follow up, 304 cups were available for analysis. The cumulative survivorship (mechanical failure as endpoint) at that time was 99.43% +/- 0.0104. Two hundred seventy-six hips were available for full clinical and radiographic review at or after the tenth anniversary. No unstable implants were noted; all implant fixation interfaces were classified as "stable bone ingrown," and the cup migration rate was zero. Based on the survivorship achieved with this implant, our results compare favorably with survivorship reported for the best cemented and cementless acetabular implant designs. PMID- 12629603 TI - Outcome in patients lost to follow-up. AB - Long-term outcome studies are valuable for providing information on the results of joint arthroplasty. However, a progressively increasing rate of attrition correlates with the length of study. The attrition rate of patients "lost to follow-up" ("non-attenders") may have a significant effect on the reported outcome. The goal of this study was to analyze the effect of the outcome in patients lost to follow-up on analysis in the setting of total knee arthroplasty. We found a lower rate of failure for revision surgery and higher satisfactory results in "non-attenders" compared with patients completing follow-up. No statistical effect of loss to follow-up care was noted on survivorship analysis when revision surgery or revision plus severe pain were considered as end points. Therefore, the patients who do not attend follow-up visits do not necessarily have poor results. PMID- 12629604 TI - Can component and patient factors account for the variance in wear rates among bilateral total hip arthroplasty patients? AB - The purpose of this study was to indirectly quantify the effect of patient and component factors on polyethylene wear in patients with bilateral hip arthroplasties. Assuming that both hips experience similar levels of activity, the confounding influence of activity on wear can be removed by comparing wear rates within subjects. We studied temporal wear patterns in 21 patients with bilateral hip arthroplasty with a mean follow-up of 102 months. Each patient had matching acetabular cup and femoral head components implanted in both hips. Regression analyses were used to assess the variation in wear rates between the first and second implanted hips. The r(2) value demonstrated that matched components and patient factors accounted for 61% of the variance in wear rates. The remaining 39% of the variance, which is unaccounted for, indicates that factors other than those related to the components and patient also play a role. PMID- 12629605 TI - A multimodality regimen for deep venous thrombosis prophylaxis in total knee arthroplasty. AB - Data indicate that deep venous thrombosis (DVT) occurs at the time of knee arthroplasty. Nevertheless, literature concerning DVT prophylaxis has only recently addressed this contention. This prospective study evaluated the efficacy of a perioperative prophylactic regimen. Between January 1996 and June 2001, 1,308 knees (964 surgeries) underwent total knee arthroplasty. Patients were treated routinely with intraoperative heparin (1000 units intravenous push before inflation of the tourniquet and 500 units at deflation), hypotensive epidural anesthesia (MAP 70-90), external pneumatic compression boots, and aspirin (325 mg, PO, BID for 6 weeks). Duplex venous ultrasonography was performed before discharge. DVT was detected in 4% of cases (1% proximal and 3% distal). Bleeding complications occurred in 1%, and perioperative medical complications occurred in 12%. PMID- 12629606 TI - Bipolar hemiarthroplasty for chronic rotator cuff tear arthropathy. AB - Massive rotator cuff tears associated with glenohumeral arthritis are currently an unsolvable clinical entity. We review our series of 14 patients with rotator cuff tear arthropathy (RCTA) who underwent a bipolar prosthesis of the shoulder. The average follow up was 27.8 months (range, 24-48 months), and the average age was 71 years (range, 57-84 years). Of these 14 patients, 9 were men and 5 women. None of these patients had undergone previous shoulder surgery, and all patients underwent conservative treatment that failed to improve their symptoms or range of motion. Preoperatively, the average active forward flexion was 30 degrees, active external rotation was 10 degrees, and the American Shoulder and Elbow Society (ASES) score was 25 points. Postoperatively, the average active forward flexion improved to 88 degrees, active external rotation increased to 37 degrees, and the ASES score improved to 80 points. Of the 14 patients, 12 stated that they had no pain with activities of daily living. Two patients continued to have moderate pain in everyday activity. PMID- 12629607 TI - Deep venous thrombosis after total hip or knee arthroplasty in a "low-risk" Chinese population. AB - Scarcely any information has been published on deep vein thrombosis (DVT) in Chinese patients after total hip arthroplasty (THA) or total knee arthroplasty (TKA). However, generally, no prophylaxis is given to patients who do not have conventional high-risk factors because they are believed to be at "low risk." We performed a prospective study on 80 such "low risk" patients undergoing THA or TKA (58 TKA and 22 THA) without prophylaxis and performed duplex ultrasonography on both lower limbs 6 to 8 days after surgery. A total of 22 patients (27.5%) showed ultrasonographic evidence of DVT. Eighteen (31%) TKAs and 4 (18.1%) THAs were complicated by DVT. Three patients showed bilateral involvement, all of whom underwent TKA. Two patients had symptomatic pulmonary embolism. The sensitivity and positive predictive value of the clinical examination was 27.2% and 31.6%, respectively. This study showed that patients who are labeled "low risk" for DVT actually had a significant risk and suggests that the current practice of providing prophylaxis to only patients deemed at "high risk" should be revised. PMID- 12629608 TI - The effect of anticoagulation on the restoration of range of motion after total knee arthroplasty: enoxaparin versus aspirin. AB - Anticoagulation used for thromboembolic prophylaxis following total knee arthroplasty (TKA) could interfere with movement. This study compares the effect of 2 anticoagulants, enoxaparin and aspirin, on restoration of range of motion (ROM) after TKA. Two groups of 75 consecutive patients, matched for age, arthritic severity, and preoperative ROM, underwent TKA. Flexion and extension milestone measures were recorded daily. Results show a highly statistically significant difference (P<.001) between the 2 groups when comparing the days on which these milestones were achieved. Group 1 (enoxaparin) reached 90 degrees, 100 degrees and 110 degrees of flexion in 8.4, 10.4, and 12.4 days, respectively. Group 2 (aspirin) reached the same goals in 6.8, 8.5, and 10.6 days, respectively. At 15 months after surgery, no statistically significant difference in flexion was seen between the groups (122 degrees vs 121 degrees ). Enoxaparin delayed the return of early but not long-term flexion after TKA. PMID- 12629610 TI - Modified iliofemoral approach for major isolated acetabular revision arthroplasty. AB - Multiple specialized exposures have been described for revision total hip arthroplasty. In this article, we report on a series of 7 patients who underwent major isolated acetabular revision surgery through a modified iliofemoral approach. The use of this approach was deemed desirable because of the individual pathology present in each case, particularly major acetabular bone loss (Paprosky 2C/3B). However, successful reconstruction was achieved in all cases with the most frequent complication being meralgia paresthetica. PMID- 12629609 TI - Embolic events during total hip arthroplasty: an echocardiographic study. AB - We performed transesophageal echocardiography on 88 patients treated with cementless or hybrid total hip arthroplasty (THA). We observed a significantly higher frequency of severe embolic events among patients without cup holes during insertion of press-fit cups (46.0%) than in patients with holes (15.6%) (P<.005). During stem insertion, severe embolic events occurred in 61.5% of hybrid operations and 5.9% of cementless operations. PaO(2), systolic blood pressure, and arterial oxygen saturation decreased significantly in severe embolic events (P<.05). The results indicate that embolic events may occur during THA whether or not cement is used. Based on these findings, we make several recommendations for technique modifications during insertion of both femoral and acetabular components in THA. PMID- 12629611 TI - Do patients need to earn total knee arthroplasty? AB - Traditional orthopaedic thinking dictates that performing total knee arthroplasty (TKA) in patients with relatively early stages of osteoarthritis (OA) will lead to a poor outcome. Our goal was to test this perception, using radiologic parameters as a marker of the degree of OA. Our study involved 130 consecutive patients who underwent TKA for OA. The radiographs of each patient were graded according to the Ahlback classification. Outcome was measured prospectively using the Oxford Knee Score (OKS), which was recorded both preoperatively and 1 year after surgery. The degree of radiologic OA changes preoperatively was not found to influence the outcome of surgery at 1 year, with all patient groups improving to a similar extent. In addition, as has been seen in previous studies, no correlation was found between symptoms of OA and radiologic appearances. The results of this study would support a re-evaluation of traditional orthopaedic practice. This conclusion coincides with other recent work that suggests that TKA is underused and is performed too late in the natural history of OA of the knee. PMID- 12629612 TI - Rapid osteolysis after revision hip arthroplasty in Paget's disease. AB - Rapid bone turnover in Paget's disease has been of concern to many surgeons performing hip arthroplasties. We present the case of a 71-year-old man with Paget's disease affecting the proximal femur. He had undergone total hip arthroplasty 14 years before. He sustained a fracture at the tip of the femoral component, which was managed with revision total hip arthroplasty. His postoperative course was complicated by rapid and profound osteolysis of the femur distal to the fracture site, secondary to disease activity. This case highlights the need for awareness of Paget's disease activity and this potential complication. PMID- 12629613 TI - A rare cause of irreducible dislocation of a total hip prosthesis. AB - This report describes a patient with an anterior dislocation of the total hip arthroplasty, which was irreducible using described techniques of closed reduction. It required an open reduction using a direct lateral approach with the release of the rectus femoris. PMID- 12629614 TI - A technique for treating periprosthetic fractures of the femur associated with deep prosthetic infection. AB - In this report a case of periprosthetic fracture of the femur associated with deep prosthetic infection after a total hip arthroplasty is treated in a novel way that allows for stabilization of the fracture and the delivery of local antibiotics. The technique allows the surgeon to stabilize the fracture and deliver antibiotics locally with a cement-coated Enders nail. This technique is useful in this combination of difficult problems as it facilitates the goals of eradicating infection, alleviating pain and improving function. PMID- 12629615 TI - Successful reimplantation of total femoral prosthesis after deep infection. AB - A 16-year-old boy developed a deep infection with methicillin-resistant Staphylococcus aureus 6 months after total femoral replacement including knee and hip joints. The deep infection was eradicated by the use of debridement with continuous irrigation and removal of all prosthetic components followed by insertion of an antibiotic-impregnated cement spacer. Six weeks later, the total femoral prosthesis could be successfully reimplanted with both acetabular and tibial components. Two and half years later, the patient remained free of infection. Debridement with continuous irrigation and an antibiotic-impregnated cement spacer can be a feasible technique for the treatment of deep infection after large prosthetic replacement. Furthermore, reimplantation should be performed after a short waiting period. Such treatment should be considered before deciding on amputation. PMID- 12629616 TI - Femoral neck ostectomy: an alternative to hip replacement in acetabular metastatic disease. AB - A 68 year old man underwent a femoral neck ostectomy as a palliative procedure for pain in the right hip secondary to an extensive acetabular metastasis from renal cell carcinoma. Excellent pain relief was achieved and he remained mobile until shortly before his death 8 months later. PMID- 12629617 TI - The histology of "reactive lines" in well-fixed components. AB - The histologic findings from 2 total knee arthroplasties (TKAs) in 1 patient who died 5 years after surgery are reported. Cement was placed under the tibial and femoral metal backs but not around the stems. All components were securely fixed. "Reactive lines," present around the tips of the stems on radiography, were seen to contain thin, soft connective tissue without debris or macrophages. The histologic features of a reactive line are described. PMID- 12629618 TI - Metal-backed patellar component failure in total knee arthroplasty presenting as a giant calf mass. AB - Failure of total knee arthroplasty (TKA) caused by wear of the polyethylene bearing surface of a metal-backed tibial platform or a metal-backed patellar component is a recognized complication. We present a case of a 78-year-old woman with a cystic mass in the left calf caused by metal wear debris from the failure of a Miller-Galante I TKA. The patient received a left TKA to treat advanced osteoarthritis in July 1990 and was lost to follow-up immediately after the operation. In December 1998, she presented at our clinic 2 days before admission, when an originally silent mass over the calf turned intolerably painful. A series of examinations revealed a calf mass caused by wear debris of total knee prosthesis and subsequent inflammation of the knee joint. Curettage of the cyst and simultaneous revision TKA were successful in relieving her symptoms. PMID- 12629619 TI - Anti-inflammatory treatment for recurrent wheezing in the first five years of life. AB - Medications identified for the treatment of recurrent wheezing in preschool children by the Expert Panel Report of the NHLBI Guidelines for the Diagnosis and Management of Asthma include inhaled corticosteroids, chromones, theophylline, and leukotriene pathway modifiers. However, these various agents differ in their mechanism, extent of action on the airway inflammatory process, and degree of clinical efficacy. Inhaled corticosteroids can control symptoms in many young children with even severe persistent wheezing, but data on their long-term safety when administered in preschool-age children are scarce. There is some information on the uninterrupted use of inhaled corticosteroids in school-age children and the absence of an adverse effect on ultimate adult height. Despite laboratory evidence of adrenal suppression in some studies, few pediatric cases of clinical adrenal insufficiency have been reported. Low-dose inhaled corticosteroid (<400 mcg/day for beclomethasone), which is adequate for controlling mild persistent symptoms, is generally safe. Chromones have a remarkable safety profile, but they are most effective for symptoms of mild severity. Promising data have been published on the efficacy and safety of leukotriene pathway modifiers when used in young children with persistent symptoms. It is uncertain whether early introduction and long-term administration of inhaled corticosteroids prevent development of irreversible airway obstruction. Nevertheless, they may be especially useful for patients with moderate to severe disease in whom other agents (chromones or leukotriene pathway modifiers) will most likely fail to control symptoms. Pediatr Pulmonol. 2003; 35:241-252. PMID- 12629620 TI - Multiresistant Pseudomonas aeruginosa in a pediatric cystic fibrosis center: natural history and implications for segregation. AB - It has been suggested that cystic fibrosis (CF) patients harboring multiresistant (MR) Pseudomonas aeruginosa (PA) should be seen in separate clinics. The aim of this study was to test the feasibility of this by longitudinally studying the consistency of isolates of MRPA in individuals. We analyzed all respiratory tract cultures undertaken in 1 year from a pediatric CF clinic population (n = 367). PA was classified as MR according to the definition of the American CF Foundation: resistance to all agents in at least two of the following groups of antibiotics: beta-lactams, aminoglycosides, and fluroquinolones. PA was cultured from 96 children during the year of study. Thirty-six were infected with at least one MR strain. Following initial identification of MRPA, MR in subsequent cultures was highly variable. Twenty-three of 36 patients had subsequent cultures in which PA was identified. However, 21 of 23 patients had at least one isolate that was not MR following detection of MRPA. The variability with time in isolation of MR strains from individuals demonstrates the potential difficulties in designing segregation policies based on antibiotic sensitivity patterns. PMID- 12629621 TI - Risk factors for initial acquisition of Pseudomonas aeruginosa in children with cystic fibrosis identified by newborn screening. AB - The objective of this study was to identify risk factors for initial detection of Pseudomonas aeruginosa (P. aeruginosa) in children with cystic fibrosis (CF) identified by newborn screening. Life history data on 180 patients, collected prospectively in a follow-up study of infants and children diagnosed with CF, were examined for factors associated with the initial detection of P. aeruginosa ascertained by oropharyngeal cultures. Univariate and multivariate Cox proportional hazards regression analyses were used to assess the effect of baseline and time-varying covariates on age at first positive culture for P. aeruginosa. Seventy-nine patients (44%) had at least one culture positive for P. aeruginosa during the study. The median age of detection was 8.1 years (95% CI, 7.0, 10.0). Median length of follow-up was 4.1 years, ranging from 0.2-15.5 years. Multivariate Cox regression analysis identified female gender (RR, 1.85; 95% CI, 1.14, 3.01), the DeltaF508 homozygous genotype (RR, 2.23; 95% CI, 1.30, 3.80), and S. aureus isolations (RR, 1.30; 95% CI, 1.11, 1.52) to be independently associated with acquisition of P. aeruginosa. Other marginally independent associations were found with days hospitalized and increased height. We conclude that female gender, homozygous DeltaF508 mutation, and S. aureus isolations are important risk factors for early P. aeruginosa detection in children with CF identified through newborn screening. PMID- 12629622 TI - Pulmonary edema of enterovirus 71 encephalomyelitis is associated with left ventricular failure: implications for treatment. AB - Epidemics of enterovirus 71 infections caused the rapid death of many children in Malaysia in 1997 and in Taiwan in 1998. Pulmonary edema occurred in most of the fatal cases and was considered to be neurogenic. The role of the heart was rarely investigated before. Between January 1998-January 2001, 34 consecutive patients who were admitted to the intensive care unit due to enterovirus infection were studied prospectively. Patients were divided into two groups: group I with pulmonary edema, and group II without pulmonary edema. Comparisons were made between the two groups based upon demographic, neurological, and cardiovascular manifestations. Group I consisted of 11 patients (5 boys, 6 girls; mean age, 22.8 months), and group II of 23 patients (12 boys, 11 girls; mean age, 28.8 months). There were no significant differences between the two groups in comparing sex, age, body weight, neurological severity, intracranial pressure, cell count, protein and glucose levels in cerebral spinal fluid, and blood pressure. All group I patients had left ventricular dysfunction, and their ejection fractions were significantly lower than those of patients in group II (37 +/- 11% vs. 75 +/ 6%, P < 0.001). Group I heart rates were higher than those of group II (175 +/- 24 vs. 137 +/- 25, P < 0.001). In group I, 9 patients who received conventional treatment died, and the only two survivors received left ventricular assist devices. In conclusion, the pulmonary edema of fulminant enterovirus 71 infection is associated with left ventricular failure. Left ventricular function is the major determinant of outcome. Early recognition of heart failure and aggressive cardiac intervention are life-saving. Pediatr Pulmonol. 2003; 35:263-268. PMID- 12629623 TI - Mesenteric lymphadenopathy as a cause of abdominal pain in children with lobar or segmental pneumonia. AB - We investigated whether mesenteric lymphadenopathy could be a cause of abdominal pain in children with lobar or segmental pneumonia. The study population consisted of 1)119 consecutive children with lobar pneumonia, older than 4 years of age, and 2) 31 healthy controls. Demographic, clinical, inflammatory, and radiographic data were recorded in all patients. All study subjects underwent abdominal ultrasound, focusing on the identification of mesenteric lymphadenopathy. One month later, a follow-up ultrasound was performed in patients with enlarged mesenteric lymph nodes at the initial examination. Forty patients complaining of abdominal pain were included in group 1, while the remaining 79 were in group 2. The two groups of patients did not significantly differ regarding their demographic, clinical (other than abdominal pain), and radiographic indices. In contrast, enlarged mesenteric lymph nodes with a sagittal diameter of at least 10 mm were identified significantly more commonly in the children of group 1 (P = 0.001). The association of enlarged lymph nodes with the presence of abdominal pain remained significant when the data were analyzed through multiple regression analysis (odds raio, 1.47; 95% confidence interval, 3-44). Enlarged mesenteric lymph nodes were found in a significantly lower ratio of healthy controls (3/31) compared to that observed in group 1 (P = 0.003). In all patients who were followed up, mesenteric lymph nodes had either decreased or were not detectable. Our findings indicate that mesenteric lymphadenopathy might be considered as a mechanism responsible for the development of abdominal pain in a relatively high percentage of children with pneumonia. PMID- 12629624 TI - Repeated blood gas monitoring in healthy children and adolescents by the transcutaneous route. AB - The aim of the study was to establish transcutaneous oxygen tension (tcPO(2)), carbon dioxide tension (tcPCO(2)), and oxygen saturation (tcSO(2)) values and to investigate the intra- and interindividual variation in these values in healthy children and adolescents. Forty-one healthy children/adolescents (21 girls; mean age, 12.1 years; SD, 3.5; range, 5.0-19.0 years) participated. Continuous recordings of tcPO(2), tcPCO(2), tcSO(2), and heart rate (HR) were made for 30 min on two occasions, with a mean interval of 10.3 days, with the child/adolescent in a sitting position. The respiratory rate (RR) was counted manually during the last 5 min of each recording. The mean values of tcPO(2), tcPCO(2), tcSO(2), HR, and RR during the last 5 min of the recordings were reported. At the first recording, the mean value for tcPO(2) was 10.8 kPa (SD 1.0), for tcPCO(2) 5.1 kPa (0.4), for tcSO(2) 97.4% (1.1), for HR 82.2 min(-1) (9.9), and for RR 19.0 min(-1) (2.8). The coefficient of variation for tcPO(2) was 9.3%, for tcPCO(2) 7.8%, for tcSO(2) 1.1%, for HR 12.0%, and for RR 14.7%. The mean intraindividual variation in tcPO(2) was 0.3 kPa (SD 1.0), tcPCO(2) 0.0 kPa (0.5), tcSO(2) 0.1% (1.3), HR 0.1 min(-1) (7.7), and RR 0.5 min(-1) (2.7). No significant differences were observed in tcPO(2), tcPCO(2), tcSO(2), HR, or RR between the two recordings. In conclusion, the reproducibility of tcPO(2), tcPCO(2), and tcSO(2) monitoring during 30 min in healthy children and adolescents was good. This indicates that transcutaneous blood gas monitoring may be used to document significant changes in blood gas tensions and oxygen saturation over time. PMID- 12629625 TI - Oxygen and ventilator weaning during inpatient pediatric pulmonary rehabilitation. AB - Rates of oxygen and ventilator weaning, and factors related to successful weaning in inpatient pediatric pulmonary programs for infants and young children, have not been frequently reported in the literature. A retrospective review was conducted of 34 infants and toddlers with either a diagnostic condition of prematurity (PM) or congenital anomalies/neuromuscular disease (CA/NM) discharged from an inpatient pulmonary program. These cases represent 67 hospital admission discharge episodes over a 6-year period. The rate of successful oxygen weaning (decrease to 0 hr per day) and ventilator weaning (decrease to <12 hr per day) and predictive factors related to successful ventilator weaning per admission discharge episode were examined. Successful oxygen weaning was achieved during 24% and successful ventilator weaning was achieved during 30% of the admission discharge episodes. No significant relationships were found between the selected demographic and clinical factors and oxygen weaning. Using a logistic regression model, the major variable associated with successful ventilator weaning per admission-discharge episode was diagnostic condition. Age at admission and the presence of comorbidities added slightly to the prediction model. The overall model yielded 86% accuracy for predicting a decrease in ventilator hours. However, projecting in which episodes children will not be weaned (negative predictive value = 88.9%) was more accurate than projecting in which episodes children will be weaned (positive predictive value = 73.3%). Although the program achieved a relatively low rate of successful ventilator weaning, children with a diagnostic condition of prematurity were more likely to be successfully weaned during inpatient pulmonary rehabilitation. PMID- 12629626 TI - Clinical characteristics of children with tracheobronchial anomalies. AB - Tracheobronchial anomalies (TBA) originate between the third and sixteenth week of gestation, and they primarily affect the main bronchi. The reported incidence varies between 1-3% of pediatric endoscopic studies. The objective of our review was to describe the clinical characteristics of patients with TBA diagnosed by flexible bronchoscopy (FB). During the period from March 1993-May 2001, we performed 580 FB at the Pediatric Services of the Catholic University Hospital. During this study period, 52 patients (9.65%) had a TBA (32 males, or 61.5%), with a mean age of 21.2 months (range, 7 days to 6 years). In 39 cases (75%), the diagnosis was made under age 2 years. Thirty-six patients with symptoms related to TBA were symptomatic (70%), and in 16 cases (30%) the TBA was an accidental finding during FB. The main clinical manifestations were recurrent atelectasis of the right upper lobe (RUL), recurrent pneumonia, or congenital stridor. Most of the TBA were localized at the RUL bronchus (47%) and were characterized by an anomaly at its origin, either agenesis or a supernumerary bronchus. Also, we found patients with tracheal bronchus and congenital tracheal stenosis. In 6 patients (12%), we had more than one TBA, the most frequent being the association between RUL and RML anomalies. In 12 cases we found another congenital airway anomaly, mainly laryngomalacia. Patients had associated diseases, such as genetic disorders in 16 cases (particularly Down syndrome), and congenital cardiopathies. In summary, in our clinical experience, TBA are more frequent than previously reported. The majority of young patients have symptoms or signs, and most are localized to the RUL. We found patients with more than one TBA, laryngomalacia associated with TBA, and at a high frequency related to Down syndrome. PMID- 12629627 TI - Good estimation of arterial carbon dioxide by end-tidal carbon dioxide monitoring in the neonatal intensive care unit. AB - End-tidal carbon dioxide pressure (PetCO(2)) was measured in the neonatal intensive care unit (NICU) to assess its reliability and accuracy in predicting arterial partial pressure of carbon dioxide (PaCO(2)). Arterial blood was drawn for gas analysis and compared with exhaled CO(2) measured by mainstream capnography. In total, 130 PetCO(2)/PaCO(2) comparisons were obtained from 61 patients (20 term and 41 preterm infants). PetCO(2) was significantly different from PaCO(2) (PetCO(2) = 42.3 +/- 10.5 mmHg vs. PaCO(2) = 45.8 +/- 12.3 mmHg, P < 0.001, mean +/- SD). The overall PetCO(2) bias (mean +/- SD) was 3.5 +/- 7.1 mmHg. There was a positive correlation between PetCO(2) and PaCO(2) (n = 130, r = 0.818, P < 0.001) in both term (n = 44, r = 0.779, P < 0.001) and preterm infants (n = 86, r = 0.849, P < 0.001). The PetCO(2) biases (95% CI) were 3.5 +/- 9.0 mmHg (0.8-6.2) in the term group and 3.4 +/- 6.0 mmHg (2.2-4.7) in the preterm group. Therefore, PetCO(2) was a valid and reliable method for monitoring PaCO(2) in neonates, especially preterm infants. This method decreases blood loss and prevents complications associated with arterial catheters. In conclusion, we recommend using mainstream capnography to monitor PetCO(2) instead of measuring PaCO(2) in the NICU. PMID- 12629628 TI - High prevalence of asthma in preschool children in Southern Brazil: a population based study. AB - The prevalence of asthma is increasing in many countries. To measure asthma's prevalence, a sample of 981 children aged 4 years old belonging to a birth cohort from 1993 was studied in Pelotas, a city in southern Brazil. A standardized questionnaire on asthma was given to the children's mothers. Information was also collected on socioeconomic level, housing conditions, genetic factors, nutritional factors, and previous infectious diseases. Current asthma (asthma diagnosed by a doctor during previous 12 months) and current wheeze (presence of wheezing during previous 12 months) were the main outcomes. The prevalence of current asthma was 18.4%, and current wheeze was 21.1%. The data were analyzed by multiple Poisson regression, and the risk factors that remained significant for both "current asthma and current wheeze" were, respectively, nonwhite color (RR = 1.41 and RR = 1.36), low maternal schooling (RR = 1.75 and RR = 1.68 for 0-4 years), history of asthma or allergy in the family (RR = 1.66 and RR = 1.85), and history of rhinitis and eczema in the child (RR = 2.11 and RR = 1.72). Male sex (RR = 1.36) and bronchiolitis (RR = 1.46) were major risk factors only for "current asthma," while smoking in pregnancy (RR = 1.30) and low birth weight (RR = 1.45) were risk factors only for "current wheeze." These results highlight the importance of asthma as a public health problem due to its high prevalence, and support the need of intervention programs against preventable risk factors. PMID- 12629629 TI - Differential cytology of bronchoalveolar lavage fluid in asthmatic children. AB - Although asthma usually begins in childhood, limited information is available as to the inflammatory reaction of asthmatic children compared to adults and the influence of age. We investigated the cytology of bronchoalveolar lavage fluid (BALF) in 39 newly diagnosed wheezy children (minimum of 3 wheezing episodes during last 6 months): 21 allergic and 18 nonallergic subjects. None had received antiinflammatory treatment. Bronchoalveolar lavage (BAL) was performed, instilling 0.5 ml.kg(-1) body weight of warmed saline in 4 successive fractions. The first 2 aliquots (BALF 1) were pooled for microbiology and cytology, and the last 2 (BALF 2) for cytology only. Recovery correlated inversely with age, the most significant being for BALF 2 (r = -0.52, P = 0.001). Children under 2 years of age had larger amounts of ciliated columnar and goblet cells (P = 0.0074). Other cell types did not show age dependency. Differential cytology was characterized by a high number of creola bodies, bronchial epithelial cells (M = 68 x 10(3).ml(-1), R = 5-349), and neutrophils (M = 92 x 10(3).ml(-1), R = 0 1,257). Eosinophils were the only cells distinguishing allergic from nonallergic subjects (P = 0.003). The 16 children with positive microbiology had more neutrophils than the noninfected (P = 0.008), the latter still having more neutrophils than found in adults. These data suggest a limited age dependency in BALF cytology. Differential cytology in BALF might be helpful in differentiating asthma in children. Neutrophil inflammation might be more important than in adults. PMID- 12629630 TI - Safety and use of sputum induction in children with cystic fibrosis. AB - We assessed the safety and use of induced sputum (IS) in children with cystic fibrosis (CF). Forty-eight children (19 males) with CF, mean age 12.6 (range, 7.3 17.0) years and median forced expired volume in 1 sec (FEV(1)) 48% (range, 14 77%) predicted were recruited. Patients spontaneously expectorated sputum and then performed sputum induction by inhalation of nebulized 7% hypertonic saline. Samples were sent for bacteriological culture, and for measurement of the following inflammatory mediators: interleukin-8, myeloperoxidase, eosinophil cationic protein, and neutrophil elastase activity. FEV(1) was performed before and after inhalation of hypertonic saline. There was no increase in mediator levels in IS compared to expectorated sputum (ES) samples. Only 3 patients demonstrated significant bronchoconstriction following inhalation of hypertonic saline, by the method used. From the ES samples, Pseudomonas aeruginosa was isolated in 13 patients, Staphylococcus aureus in 7 patients, Stenotrophomonas maltophilia in 1 patient, and both Pseudomonas aeruginosa and Staphylococcus aureus in 5 patients. All these organisms were found in the IS samples. However, in 2 patients whose ES grew no organisms, one patient's IS grew Pseudomonas aeruginosa, and the other patient's IS grew Staphylococcus aureus. In our study, sputum induction was safe, with no proinflammatory effect. PMID- 12629631 TI - Laryngotracheal involvement of relapsing polychondritis in a Korean girl. AB - We describe a young girl presenting with acute dyspnea because of laryngotracheal involvement of relapsing polychondritis (RP). RP, a multisystemic disorder of unknown etiology, is a very rare disease in children. It is characterized by inflammation and destruction of the cartilaginous structure of many organs, including the respiratory tract. Early respiratory tract involvement in younger patients is the greatest threat to life, and aggressive therapy, including tracheostomy and intravenous high-dose steroids, is advocated. PMID- 12629632 TI - Pediatric pulmonary carcinoid: a case report and review of the literature. AB - We report on the youngest known patient with a pulmonary carcinoid, a rare tumor in children. We review the management of and prognosis for this tumor. Diagnosis can be delayed because of low clinical suspicion and the many ways in which these tumors can present. Treatment is surgical and geared towards complete resection while sparing healthy lung parenchyma. Prognosis is generally good, but careful long-term follow-up is needed for potential recurrences or associated tumors. PMID- 12629633 TI - Late-onset respiratory distress after inhalation of laundry detergent. AB - Accidental poisoning with household cleaning products can pose significant risks to children. Exposure to granular laundry detergents accounts for a number of calls each year to poison control centers, though few of these exposures result in hospitalization. While caustic gastrointestinal injury resulting from ingestion of these highly alkaline cleaning agents is well-recognized, few reports address the potential damage to the respiratory tract that can occur following ingestion or inhalation of granular laundry detergent. We present a previously healthy 1-year-old who presented to the emergency department with Late onset stridor and increased work of breathing following presumed inhalation of granular laundry detergent. Parents, primary care providers, and emergency department physicians need to be aware of the potential toxicity of these widely used household products. PMID- 12629634 TI - To the editor: re: Brown KA, Platt R, and Bates JHT, "Automated analysis of paradoxical ribcage motion during sleep in infants," Pediatr Pulmonol 2002;33:38 46. PMID- 12629636 TI - Mutational patterns of paired blood and rectal biopsies in HIV-infected patients on HAART. AB - Blood and concurrent rectal biopsy samples of human immunodeficiency virus type 1 (HIV-1)-positive highly active antiretroviral therapy (HAART)-treated patients were tested for genotypic resistance by direct sequencing of reverse transcriptase (RT) and protease (PR) regions to compare the patterns of resistance in these compartments. Fourteen subjects (five with undetectable plasma viral load (pVL) and nine persistently viremic) were studied. Four of five patients with undetectable pVL also had undetectable mucosal HIV RNA; sequence analyses from proviral DNA (PBMCs and rectal biopsy) were obtained with none or few resistance-associated mutations and no alteration of susceptibility profile. All viremic patients, and one with negative pVL, had detectable levels of mucosal HIV RNA (1.93-4.21 log(10) copies/mg); sequences of HIV RNA (plasma and/or rectal biopsy) were also obtained, and multiple mutations generally compatible with current/past medications were detected. Overall, 40 HIV-1 PR and 42 RT sequences were analyzed, yielding a total of 42 PR and 47 RT sequence pairs (plasma/tissue RNA; plasma-RNA/tissue-DNA; PBMC/tissue-DNA; tissue-DNA/RNA; tissue-RNA/PBMC-DNA; PBMC-DNA/plasma-RNA), which almost always differed at the total amino acid level (median percentage discordance 8.08% in the PR, 4.8% in RT). The median percentage of resistance position discordance equaled 88.8% (IQR = 20-100) in the PR and 74.55% (IQR = 31.75-100%) in the RT pairs, respectively. Different resistance levels were detected by means of a computer-assisted interpretation of mutational profiles. The results support the multiform evolution of HIV genotype in various body compartments and emphasize the participation of intestinal mucosa in HIV genotype selection. Samples from diverse tissues should be used for resistance evaluation to obtain a complete picture of drug resistance for antiretroviral-treated patients. PMID- 12629637 TI - Introduction of HIV type 1 non-B subtypes into Eastern Andalusia through immigration. AB - A study of the distribution of HIV-1 subtypes in the native and immigrant populations of Eastern Andalusia (Southern Spain) was conducted to determine any changes between 1983 and 2001 and to identify antiretroviral resistance mutations in non-B subtype strains among the immigrant population. The study included 111 native patients from Eastern Andalusia: 94 infected with HIV before 1996 and 17 infected since 1996. A parallel study was conducted on 26 HIV-positive immigrants from Africa. Subtyping was done with the heteroduplex mobility assay. Resistance mutations were determined by line probe assay. A total of 137 patients were studied: 9.2% had subtype A (n = 12), 80.8% subtype B (n = 105), and 1.5% subtype C (n = 2). Among the Eastern Andalusia population infected before 1996, 10.9% had non-B subtypes, compared with 23.5% of those infected after that year. The greatest percentage of non-B subtypes (52.4%) was found among the immigrant population. Resistance mutation K70R was detected in one of the six immigrants with non-B subtype and M41L in another. There has been a slight increase in the diversity of HIV-1 subtypes in Eastern Andalusia over the past few years, possibly influenced by non-B subtypes introduced by immigrants from sub-Saharan Africa. PMID- 12629638 TI - Identification of a unique BK virus variant in the CNS of a patient with AIDS. AB - Human polyomavirus BK (BKV; GenBank or EMBL or DDBJ accession no. NC001538) is often reactivated in immunosuppressed patients. Reactivation has been associated primarily with excretion of the virus in the urine, and there have been few reports of renal and/or neurological disease caused by BKV in patients with acquired immunodeficiency syndrome (AIDS). Polymerase chain reaction, Southern blotting, and sequencing were used to detect and identify the noncoding control region (NCCR) of BKV in different tissues in an AIDS patient with meningoencephalitis, retinitis, and nephritis. An undescribed reorganized NCCR variant of the virus, completely different from the variants detected in peripheral blood leukocytes (PBLs) and urine, was identified in the cerebrospinal fluid (CSF) and CNS tissues. These results suggest that rearrangements in the NCCR of the virus have resulted in a BKV variant, which is better adapted to the host cell machinery of the cells in CNS tissue. The rearranged variant (BKV CNS) might have been involved in the initiation and/or development of the pathological lesions observed in the CNS-related tissues of this patient. PMID- 12629639 TI - Molecular and serological aspects of HBsAg-negative hepatitis B virus infections in North America. AB - A few hepatitis B virus (HBV) infections are characterized by the presence of HBV DNA in serum or liver tissue, or both, in the absence of detectable hepatitis B surface antigen (HBsAg) in serum. However, such infections have rarely been described previously in North American patients. In the present study, 31 hepatocellular carcinoma (HCC) patients from the United States and Canada who had no detectable HBsAg in their serum were studied. In these 31 HBsAg-negative HCC patients, HBV DNA was detected in HCC and/or in adjacent nontumorous liver tissue using nested polymerase chain reaction (PCR) in 5/9 (56%) patients from the United States and in 12/22 (55%) from Canada. The 17 HBV DNA-positive/HBsAg negative patients from the United States and Canada included 9 without any serological markers for HBV and 8 with detectable antibodies to hepatitis B core antigen. In these patients, HBV genotype C was the most prevalent genotype (11/17; 64%). HBV genotypes have not been previously reported in HCC patients from North America. Replicative intermediate forms of HBV (covalently closed circular HBV DNA) were detected in 2/17 (12%) HBV DNA-positive/HBsAg-negative patients, indicating that at least two of these patients had actively replicating HBV infections. The use of tests to detect HBV DNA permitted the identification of HBV infections in HBsAg-negative HCC patients from North America. Among these patients, those with antibody to hepatitis C virus (HCV) would otherwise have been designated "HCV-associated HCCs" based on serological tests alone. These findings provide a new perspective on determining the possible viral etiologies of HCCs in North America. PMID- 12629640 TI - Total HCV core antigen assay: a new marker of hepatitis C viremia for monitoring the progress of therapy. AB - The ability of the total hepatitis C virus (HCV) core antigen assay was evaluated for monitoring the therapeutic responses of HCV-infected patients treated with interferon. The ability to detect and quantitate an independent structural protein component of HCV, in the presence of circulating antibodies, makes this assay a valuable new tool in diagnosis and treatment monitoring. Measurement of total core antigen showed a strong dynamic correlation with HCV RNA data and may serve as an alternative direct marker of viral infection. In addition, with the advent of additional treatment protocols, a rapid, reliable assay for changes in HCV load may permit more frequent patient assessment and tailoring of the therapeutic regimen. PMID- 12629641 TI - Suggested role of the Golgi apparatus and endoplasmic reticulum for crucial sites of hepatitis C virus replication in human lymphoblastoid cells infected in vitro. AB - Iacovacci et al. [(1997a) Research in Virology 148:147-151] described that the euploid diploid cells, of the normal human bone marrow-derived lymphoblastoid B cell line TO.FE., are susceptible to hepatitis C virus (HCV) infection and support long term virus production. Transmission electron microscopy described some steps of HCV replication cycle in this in vitro infected cellular system [Serafino et al. (1997) Research in Virology 148:153-159]. In the present study, in order to identify the intracellular sites involved in HCV replication, the ultrastructural changes associated with infection in TO.FE. cells were correlated with the subcellular localisation of structural and nonstructural viral proteins. Transmission electron microscopy and confocal microscopy data indicate that these viral proteins appeared located in the Golgi apparatus and endoplasmic reticulum, suggesting an active involvement of these compartments in viral assembly and morphogenesis. Furthermore, transmission and scanning electron microscopic observations on cultures infected chronically support the hypothesis that these cellular compartments may serve as starting sites of the morphological changes associated to viral infection and replication, leading to cell-cell fusion, syncytia formation, and finally lysis of infected cells and virus release. PMID- 12629642 TI - High-density lipoprotein binding rate differs greatly between genotypes 1b and 2a/2b of hepatitis C virus. AB - The hepatitis C virus (HCV) virion is associated with lipoproteins and immunoglobulins in the sera of patients with chronic hepatitis C; however, an accurate binding rate of HCV to lipoproteins or immunoglobulins has not yet been elucidated. Therefore, the accurate binding rate of HCV to low-density lipoproteins (LDL), high-density lipoproteins (HDL), and immunoglobulins was measured quantitatively by a real-time PCR assay. The immunoglobulin binding rate of HCV was found to be greater than 97.5% in most patients, as compared with an LDL binding rate of greater than 80% in most patients. In contrast, the HDL binding rate was greater than 98% in the genotype 2a/2b patients, while it varied in the genotype 1b patients. The genotype 2a/2b HCV not only had a higher LDL binding rate but also had a strikingly higher HDL binding rate than that of the genotype 1b HCV. These lipoprotein binding rates correlated neither to any patient's variables, including the serum apolipoprotein levels, nor to the viral load or the hypervariable region 1 (HVR 1) amino acid sequences. Most of the HCV virions in the sera of such patients have been shown to be associated simultaneously with immunoglobulins and LDL and/or HDL, but not exclusively. PMID- 12629643 TI - Assessment of iatrogenic transmission of HCV in Southern Italy: was the cause the Salk polio vaccination? AB - Since the first studies on hepatitis C virus (HCV) prevalence were published, it has been evident that southern Italy is an area of hyperendemicity. A recent study conducted in southern Italy suggested that the high prevalence of HCV infection might be the result of past iatrogenic transmission. Polio vaccination with the parenteral Salk vaccine between 1956 and 1965 by multiple use of unsafe glass syringes may have been one of the major causes of the spread of HCV infection among southern Italian adults who are now older than 40 years of age. Persons born between the 1940s and early 1960s have a nearly 3-fold increased risk of HCV seropositivity than the younger age group. The findings are consistent with a cohort effect of exposure to the Salk parenteral vaccination. PMID- 12629644 TI - Analysis of the CD8-positive T cell response in Japanese patients with chronic hepatitis C using HLA-A*2402 peptide tetramers. AB - Hepatitis C virus (HCV)-specific CD8+ cytotoxic T lymphocytes (CTL) contribute to viral clearance in acute, self-limited hepatitis C as well as to liver cell injury in the more frequent cases with chronic hepatitis C. Although HLA class I peptide tetramers have been used to detect circulating HCV epitope-specific CTL with a high sensitivity and specificity, this technique has been targeted exclusively to the most frequent HLA haplotypes in the Caucasian population and the large number of HCV-infected Asian patients, most of whom are HLA-A24 positive, have not been studied. The current study determines the frequency, phenotype, and clinical significance of HCV-specific CD8(+) T lymphocytes with five different HLA-A*2402 tetramers in 43 HCV infected Japanese patients and 32 controls. Overall, tetramer(+) cells were detected in the blood of 33 of 43 patients at frequencies of 0.064-0.75% CD8(+)CD4(-)CD14(-)CD19(-) T lymphocytes. Interestingly, although the T cell response was always targeted multispecifically against epitopes in different HCV proteins, the relative frequency of cells stained with individual tetramers differed between patients. Furthermore, tetramer(+)CD8(+) T lymphocytes were highly activated, but the phenotypes of different tetramer(+) cells varied in each patient. In conclusion, HLA-A24 restricted, HCV-specific CD8(+) T lymphocytes are found at similar frequencies in Asian patients as HLA-A2 restricted, HCV-specific CD8(+) T lymphocytes in Caucasian patients. Differences in the frequency and activation status of individual tetramer(+) cell populations suggest that CD8(+) T lymphocytes with different HCV epitope specificity may mediate differential pathogenetic effects in chronic hepatitis C. PMID- 12629645 TI - Hepatitis C virus E2 and NS5A region variability during sequential treatment with two interferon-alpha preparations. AB - To determine the pattern and significance of the HCV genetic heterogeneity before and during treatment with recombinant-2b or lymphoblastoid alpha-interferon, hypervariable region 1 (HVR-1) and NS5A quasispecies were characterised by cloning and sequencing in 12 HCV-1b-infected subjects. Patients were either responder-relapsers or non-responders to treatment. Extensive amino acid sequence analysis was applied to reveal the significance of HCV variation at key sites within HVR-1 and NS5A regions. Genetic complexity, genetic diversity, and the non synonymous to synonymous substitution ratios of HVR-1 quasispecies decreased during treatment in responder-relapser patients only, and more markedly so following lymphoblastoid alpha-interferon. In non-responders, the HVR-1 quasispecies broadened. Amino acids G406 and Q409, which represent a major viral epitope, were highly conserved throughout treatment. Responder-relapser patients had a higher mutation frequency in NS5A than non-responders. Lymphoblastoid alpha interferon promoted the selection of intermediate Interferon Sensitivity Determining Region (ISDR) sequences, whereas recombinant-2b alpha-interferon favoured maintenance or selection of conserved ISDR sequences. Variability upstream of the ISDR was associated with treatment response, but the amino acid substitutions conferring higher replicative ability to in vitro HCV replicons were absent in in vivo isolates. In conclusion, the pattern of HVR-1 quasispecies evolution correlates with the clinical response, and the conservation of specific amino acids may be useful for immune targeting in vivo. In responder-relapser patients, the initial HVR-1 evolution resembles that found in sustained responders. Variability within the entire NS5A, as opposed to a single region (ISDR), may have a role in influencing alpha-interferon treatment outcome. A differential effect of different alpha-interferon preparations on HCV quasispecies kinetics may exist. PMID- 12629646 TI - Molecular epidemiological and clinical aspects of hepatitis D virus in a unique triple hepatitis viruses (B, C, D) endemic community in Taiwan. AB - The molecular epidemiological and clinical aspects of hepatitis D virus (HDV) in a unique HBV, HCV, and HDV triple virus endemic community in southern Taiwan were investigated. A total of 2,909 residents aged 45 or older were screened for hepatitis B surface antigen (HBsAg), anti-HCV antibody, and anti-HDV antibody (specifically for HBsAg-positive carriers). Factors that might be associated with HDV infection, viral nucleic acid detection, and genotyping of HBV, HCV, and HDV were investigated. The prevalence of HBsAg and anti-HCV were 12.6% (366/2,909) and 41.6% (1,227/2,909), respectively. For HBsAg carriers, 15.3% (56/366) were positive for anti-HDV assay. Living in a higher endemic district of HCV infection (odds ratio [OR] = 3.2; 95% confidence interval [CI] = 1.7-6.3), male gender (OR = 1.9; 95% CI = 1.1-3.6) and co-infection with HCV (OR = 1.8; 95% CI = 1.0-3.3) were significantly independent factors associated with HDV infection. The detection rate of HDV RNA among anti-HDV-positive patients was only 12.7% (7/55). The mean HBV titer of triple infection group was significantly lower than in the HBV/HDV co-infection group (2.23 vs 3.05 in log(10), copies/ml, P = 0.046). HCV RNA detection among the triple infection group showed 47.4% (9/19) viremia rate and viral loads of 579,121 IU/ml in median (16,803-1,551,190 IU/ml). The prevalent genotype of HBV was type B (23/25); HCV was 1b (7/9) and HDV was IIa/IIb (4/4). Only the presence of HCV RNA predicted the presence of elevated ALT significantly (OR = 25.0; 95% CI = 3.39-184.6). In conclusion, the geographical aggregation of HDV infection paralleled that of HCV infection in this community. HCV suppressed the replication of HBV among triple vital infection patients. HBV and HDV lapsed into a remission or nonreplicative phase in most cases, and HCV acted as a dominant factor in triple viral-infected individuals. Only the presence of HCV RNA was associated with elevated ALT values, but not HBV or HDV. PMID- 12629647 TI - Recombinant murine L20B cell line supports multiplication of group A coxsackieviruses. AB - Sensitive, reliable, and rapid methods of virus culture are essential for wild poliovirus isolation and identification from stool specimens collected from cases of acute flaccid paralysis. Recently, recombinant murine cell lines expressing human poliovirus receptor (CD155) on the cell surface have become available. These cells are sensitive selectively to poliovirus because the poliovirus receptor is not used by other enteroviruses. In early field studies non-polio enteroviruses were not isolated from stool samples of cases of acute flaccid paralysis using L20B cells. For the past 3 years, L20B cells were used extensively in our laboratory for virus culture. The objective of the present study was to identify non-polio enteroviruses causing cytopathic changes in L20B cells. Stool specimens collected from 1,153 cases of acute flaccid paralysis and 2,670 apparently healthy children were tested for enteroviruses using RD and L20B cell lines. A small number of viruses other than poliovirus causing cytopathic effect in L20B cells were isolated. Such isolates detected in other polio network laboratories in India were also included. The virus isolates were typed using partial VP1 nucleotide sequence analysis and virus neutralization tests. Of the 111 viruses studied, 8 were non-enteroviruses. Among the 103 non-polio enteroviruses, 73 were identified as group A coxsackieviruses. Of the 30 isolates that could not be characterized, 1 remained unidentified even by sequence analysis and 29 did not reach high titers in L20B as well as RD cells. In conclusion, coxsackie A viruses multiply in L20B cells causing cytopathic effect. Coxsackievirus A8 and coxsackievirus A10 were predominant among the eight coxsackie A virus types so far identified. PMID- 12629648 TI - Enhancement of humoral immune responses to a human cytomegalovirus DNA vaccine: adjuvant effects of aluminum phosphate and CpG oligodeoxynucleotides. AB - A human cytomegalovirus (HCMV) glycoprotein B (gpUL55) DNA vaccine has been evaluated in BALB/c mice. Intramuscular immunization of these mice with pRc/CMV2 gB resulted in the generation of high levels of gpUL55-specific antibody (geometric mean titer [GMT] 1:8900) and neutralizing antibody (GMT 1:74) after 2 booster doses given 5 and 10 weeks after primary inoculation. Emulsifying the construct with the aluminum phosphate gel adjuvant Adju-Phos before immunization enhanced gpUL55-specific antibody responses (GMT 1:17800, P = 0.04). Co immunization with CpG oligodeoxynucleotides was shown to enhance levels of neutralizing antibodies generated by immunization of mice with a pRc/CMV2-gB/Adju Phos emulsion (P = 0.04). The results provide a rationale for evaluating combinations of other HCMV proteins for incorporation into a multi-target DNA vaccine, and for the optimization of adjuvant usage, to elicit enhanced levels of neutralizing antibodies. 2003. PMID- 12629649 TI - Elevated levels of total and dengue virus-specific immunoglobulin E in patients with varying disease severity. AB - The kinetics of total and dengue virus-specific immunoglobulin E (IgE) were studied in serial serum samples obtained from 168 patients, 41 of whom suffered from primary dengue virus infection and 127 suffered from secondary dengue virus infection. Seventy-one patients were classified as dengue fever, 30 as dengue hemorrhagic fever, and 67 as dengue shock syndrome. A control group included single serum samples from patients with a herpes virus infection (n = 14), non dengue febrile patients (n = 10), and healthy blood donors (n = 10). Patients with dengue virus infection had higher levels of total and dengue virus-specific IgE than non-dengue patients (P < 0.05). Patients with secondary dengue virus infections had not significantly increased levels of both total and dengue virus specific IgE in the acute phase of disease compared to patients with primary dengue virus infections. Dengue virus-specific IgE was significantly higher in dengue hemorrhagic fever and/or dengue shock syndrome patients compared to dengue fever and non-dengue patients (P < 0.05). In conclusion, this study showed elevated total and dengue virus-specific IgE serum antibody levels in the acute stage of disease. Therefore, measurement of both total and dengue virus-specific IgE serum antibodies can be used as an additional prognostic marker in the development of severe complications in dengue virus infections. In addition, the presence and increase of dengue virus-specific IgE serum antibodies in patients with dengue virus infections is suggestive of the pathogenetic role that IgE may play in the hemostatic disorders observed in dengue hemorrhagic fever and dengue shock syndrome. PMID- 12629650 TI - Variation in enterovirus receptor genes. AB - The increased incidence of a enterovirus infections observed in patients with type 1 diabetes preceding the development of the clinical disease could be partially explained by variation in the genes coding for enterovirus receptors. We carried out sequence analysis of the most common enterovirus receptor molecules in 21 diabetic children and 20 healthy adults. DNA was isolated from the leukocytes, and gene regions known to code for virus-recognizing domains in major enterovirus receptors were amplified and sequenced. Heterozygous single nucleotide polymorphism (SNP), Ala 67 (GCG) --> Thr (ACG), was detected in the poliovirus receptor gene in four individuals in the diabetes group, but not in the control group. However, serological studies could not confirm that this substitution would convey different susceptibility to poliovirus infection. A heterozygous SNP, Lys 29 (AAG) --> Met (ATG), was found in the intracellular adhesion molecule-1 (ICAM-1) (receptor for rhinoviruses and some coxsackie A viruses) in one individual in both groups. A silent SNP in the alpha2 integrin subunit gene (echovirus 1 receptor) was frequently found in both groups, a silent heterozygotic SNP in coxsackievirus-adenovirus receptor (coxsackie B virus receptor) gene was seen in one individual in the diabetes group, whereas no variation was found in the DAF (echovirus receptor) and beta3 integrin subunit sequences (receptor for coxsackievirus A9) studied. In conclusion, both synonymous and nonsynonymous sequence variability of genes coding for enterovirus and rhinovirus receptors was shown to occur, but no pattern directly specific for type 1 diabetes was found. = PMID- 12629651 TI - Comparative study of enterovirus 71 infection of human cell lines. AB - The cell tropism of enterovirus 71 (Enteroviridae) in neuronal, glial and laryngeal cells. The 4643 strain, an enterovirus 71 isolate from a patient in Taiwan, was used to infect three human cell lines representing neuronal cells (SK N-SH, neuroblastoma), glial cells (U373MG, glioblastoma), and laryngeal cells (HEp-2, larynx epidermoid carcinoma). Immunofluorescent staining and transmission electron microscopy (TEM) were used to detect mature enterovirus 71 4643 virions in these cell lines. The three cell lines were also compared for presence of virus-mediated cytopathic effect (CPE), synthesis of infected cell-specific proteins, viral (-) RNA, and virus replication rate. Virus particles were detected by TEM, and viral replication increased over time, indicating the existence and release of mature viruses from all three infected cell lines. The most severe CPE and the highest viral replication rate were observed in the SK-N SH cells. Further screening of the infected cell lines by microarray analysis revealed that the neuron growth factor receptor (NGFR) gene was uniquely upregulated in infected SK-N-SH cells, implying that the receptor encoded by this gene may be involved in cell tropism. The data show that neurons are vulnerable to enterovirus 71 4643 infection and are consistent with the clinical observation that enterovirus 71 4643 targets mainly neuronal cells but is also found in many organs in conjunction with an inflammatory reaction. PMID- 12629652 TI - Inhibition of enterovirus 71-induced apoptosis by allophycocyanin isolated from a blue-green alga Spirulina platensis. AB - Enterovirus 71 infection causes significant morbidity and mortality in children, yet there is no effective treatment. In this study, a protein-bound pigment, allophycocyanin purified from blue-green algae is first reported to exhibit anti enterovirus 71 activity. Allophycocyanin neutralized the enterovirus 71-induced cytopathic effect in both human rhabdomyosarcoma cells and African green monkey kidney cells. The 50% inhibitory concentration of allophycocyanin for neutralizing the enterovirus 71-induced cytopathic effect was approximately 0.045 +/- 0.012 microM in green monkey kidney cells. The cytotoxic concentrations of allophycocyanin for rhabdomyosarcoma cells and African green monkey kidney cells were 1.653 +/- 0.003 microM and 1.521 +/- 0.012 microM, respectively. A plaque reduction assay showed that the concentrations of allophycocyanin for reducing plaque formation by 50% were approximately 0.056 +/- 0.007 microM and 0.101 +/- 0.032 microM, when allophycocyanin were added at the state of viral adsorption and post-adsorption, respectively. Antiviral activity was more efficient in cultures treated with allophycocyanin before viral infection compared with that in the cultures treated after infection. Allophycocyanin was also able to delay viral RNA synthesis in the infected cells and to abate the apoptotic process in enterovirus 71-infected rhabdomyosarcoma cells with evidence of characteristic DNA fragmentation, decreasing membrane damage and declining cell sub-G1 phase. It is concluded that allophycocyanin possesses antiviral activity and has a potential for development as an anti-enterovirus 71 agent. PMID- 12629653 TI - Epstein-Barr virus infection of rat lymphocytes expressing human CD21 results in restricted latent viral gene expression and not in immunoblastic transformation. AB - Transgenic rats expressing human CD21 gene (hCD21) driven by the mouse immunoglobulin enhancer were generated. hCD21 was expressed in lymphoid tissues, especially in the spleen and in the brain. Flow cytometric analysis indicated that about 20% of spleen cells, most having a B-lymphocyte marker, expressed hCD21. After Epstein-Barr virus (EBV) infection of spleen cells, EBV-determined nuclear antigen (EBNA) was first detected on Day 4 and reached a maximum of 0.3% on Day 5, but the infection was abortive and was not followed by blastogenesis, cellular DNA synthesis or proliferation. Reverse transcription-polymerase chain reaction (RT-PCR) analyses demonstrated that EBV-infected spleen cells expressed EBNA1 and EBV-encoded small RNA (EBER), but not other latent EBV products. EBNA promoter analysis by RT-PCR indicated that the Q promoter was active, whereas C and W promoters were not active. The present findings indicate that human and rat lymphocytes respond to EBV infection differently in vitro. PMID- 12629654 TI - Absence of SLAM mutations in EBV-associated lymphoproliferative disease patients. AB - X-linked lymphoproliferative disease is a rare inherited immunodeficiency in which affected males present abnormal responses to Epstein-Barr virus (EBV) infection. The gene defective in X-linked lymphoproliferative disease, SH2D1A (also named SAP or DSHP), has been identified and shown to code for an adapter protein that interacts with signaling lymphocytic activation molecule (SLAM) and several other members of the CD2 superfamily. SH2D1A is mutated in no more than 60% of X-linked lymphoproliferative disease patients. It could be postulated that a certain percentage of patients without apparent maternal transmission might be caused by other gene(s) in SH2D1A-related signal transduction pathways. Being a partner of SH2D1A and having a key role in proliferation and differentiation of the T- and B-lymphocytes, SLAM was considered as a candidate gene for patients who manifest symptoms of X-linked lymphoproliferative disease but who have no mutations in SH2D1A. As a first step, SLAM mutations were screened for from cDNA of the lymphoblastoid cell line of all available patients. Then conditions for PCR, single-strand conformational polymorphism (SSCP), heteroduplex analysis, and sequencing were established in all eight exons of SLAM. A total of 31 typical and atypical patients were analysed, from which six novel nucleotide variants were identified; however, none of these variants seems to cause abnormal function of the SLAM gene. Therefore, mutations in coding regions or splicing sites of SLAM are unlikely to play a major role in the mechanism of EBV-associated lymphoproliferation. PMID- 12629655 TI - Dynamics of Epstein-Barr virus load in pyothorax-associated lymphoma. AB - We treated a 73-year-old man who developed a pyothorax-associated pleural lymphoma after a 52-year history of tuberculous pleuritis. The lymphoma was classified histologically as a diffuse large, B-cell type. Epstein-Barr virus (EBV) was identified in the tumor by immunocytochemical and molecular methods. Chemotherapy both without and with the addition of rituximab was only minimally effective, but adding radiotherapy to chemotherapy reduced the tumor size, resulting in a partial remission. The EBV load measured by a quantitative real time polymerase chain reaction correlated well with the tumor size and the serum lactate dehydrogenase concentration. Monitoring the EBV load may be useful in the management of pyothorax-associated lymphoma. PMID- 12629656 TI - GB virus C genotype determination in GB virus-C/HIV co-infected individuals. AB - Several recent studies have indicated that patients infected with human immunodeficiency virus (HIV) exhibit a beneficial effect of co-infection with GB virus C (GBV-C). The benefit is demonstrated by slower progression to acquired immunodeficiency syndrome (AIDS) and prolonged survival time after the development of AIDS. In some but not all studies, a significant association between GBV-C/HIV co-infection and increased CD4(+) cell counts has been reported. To understand further the possible role that GBV-C might play in the reduced morbidity and mortality among HIV-infected patients, we sought to examine the presence of different GBV-C genotypes in a cohort of co-infected patients. PCR products derived from the 5'-untranslated region (5'-UTR) and the second envelope gene (E2) were sequenced directly and genotyped by phylogenetic analysis. While 5'-UTR analysis delineated the major type, analysis of the complete E2 gene was required for identification of group 2 subtypes, designated 2a and 2b. Among 35 patients tested, GBV-C genotype was determined for 33: two patients were infected with genotype 1, 12 with type 2a, and 19 with type 2b. Clinical data were available for 25 genotyped patients: one infected with genotype 1, nine with genotype 2a, and 15 with type 2b. CD4 cell counts tended to be lower in patients infected with genotype 2a compared with those with genotype 2b (310 +/- 136 vs 430 +/- 199, P = 0.054). Additional studies with larger cohorts from separate geographical regions are needed to determine whether a particular GBV-C genotype is associated with reduced morbidity or mortality among HIV co-infected patients. PMID- 12629657 TI - Prevalence of GBV-C infection among dental personnel. AB - Healthcare workers who carry out exposure-prone procedures are theoretically at increased risk of acquiring blood-borne virus infections. GB virus C (GBV-C) is a recently described blood-borne virus that is related distantly to hepatitis C virus. The occupational risk of GBV-C infection to healthcare workers is unknown. This study collected detailed occupational and personal risk data in parallel with a blood specimen, to establish the prevalence and determinants of GBV-C infection among dental healthcare workers. The presence of GBV-C antibodies was detected using commercially available ELISA; GBV-C RNA was detected by nested PCR using primers from the conserved 5' noncoding region. The overall prevalence of GBV-C antibodies among the study population was 11.1% (98/880, 95% confidence interval [CI], 9.1-13.4%) and 4.6% were positive for GBV-C RNA (46/879, 95% CI, 2.5-5.1%), resulting in a cumulative prevalence of 15.7%. These figures are similar to those described in other populations. There was no significant difference in lifetime exposure to GBV-C between dentists (17.7%) and dental nurses/hygienists (14.3%). Significantly more dental nurses/hygienists aged 16-30 years had been exposed to GBV-C compared to dentists of the same age (chi(2) = 13.75; P < 0.001). Conversely, significantly more dentists 46 years or older had evidence of exposure to GBV-C compared to dental nurses/hygienists (chi(2) = 6.79; P = 0.009). The high prevalence of GBV-C infection did not seem to be related to past parenteral exposure, and the data suggest that sexual transmission, rather than occupational transmission, was a more important route for GBV-C infection among this population. PMID- 12629658 TI - GB virus C epidemiology in Denmark: different routes of transmission in children and low- and high-risk adults. AB - With the demonstration of an effect of GBV-C infection on the outcome of HIV infection, it has become important to understand the epidemiology of GBV-C. The purpose of this study was to determine the prevalence in high- and low-risk populations. The following populations were tested: school children, 9 and 15 years of age (n = 901), blood donors (n = 5,203), hospital employees (n = 1,432), and prisoners and injecting drug users (n = 447). In-house RT-PCR for GBV-CRNA was used together with a commercial ELISA for anti-E2 (Boehringer, Germany). In addition, questionnaires for risk factors for transmission and serological tests for HIV and hepatitis were applied. The overall prevalence of GBV-CRNA was 1.4% among children, 2.2% among blood donors, 2.2% among hospital employees, 12.5% among non-injecting prisoners, and 34.9% among drug injectors. Correspondingly anti-E2 was found in 0.3%, 12.3%, 25.0%, and 42.7%. Among hospital employees, independent risk factors for GBV-C were professions with blood exposure and sexual risk partners. Among prisoners and drug users, injecting and a sexual risk index were associated independently with GBV-C. Based on these results, the following hypothesis is suggested: GBV-C is transmitted frequently at birth or early childhood and this leads to chronic infection in most cases. Sexual transmission is the most important route of transmission in the adult population but this infection is usually transient. Blood borne transmission plays a role among health care workers and injecting drug users and GBV-C should be further evaluated as a surrogate marker for professional blood exposure. PMID- 12629659 TI - Herpes simplex virus type 1 can either suppress or enhance human immunodeficiency virus type 1 replication in CD4-positive T lymphocytes. AB - It was proposed recently that CEM CD4-positive T cells infected chronically by herpes simplex virus type 1 (HSV-1) and human immunodeficiency virus type 1 (HIV 1) (CEM(HSV/HIV)) may be used as a model for studying HIV/HSV interactions. To ascertain whether HSV-HIV coinfection of T lymphocytes has a role in promoting progression of lentiviral infection, T cells infected chronically by either HSV-1 (CEM(HSV)) or HIV-1 (CEM(HIV)) were challenged with a superinfecting dose of HIV 1 or HSV-1. The results show a positive influence on HIV growth when CEM(HIV) cells were superinfected with HSV-1 to an extent that was dependent on the multiplicity of superinfection used. In contrast, HIV superinfection of CEM(HSV) cells resulted in a delay of HIV-1 production and in a lack of HSV-mediated LTR transactivation. These effects were due to cell growth inhibition and apoptosis, resulting from persistent HSV-1 infection. Treatment of CEM(HSV) with acyclovir inhibited completely the HSV-1 cytopathic effects and allowed efficient HIV-1 replication. These data may be relevant in clarifying the role of HIV/HSV interaction in the pathogenesis of AIDS. PMID- 12629660 TI - Comparative evaluation of measles virus-specific RT-PCR methods through an international collaborative study. AB - Comparison of RT-PCR assays established in house at various places revealed that laboratories could differ in sensitivity by as much as 1,000-fold in terms of the ability to detect measles virus sequences in clinical samples. The study indicates that PCR findings, positive or negative, are questionable if they are not supported by the associated data demonstrating the overall sensitivity of the assay applied. Measles virus-specific RT-PCR-based assays need to be validated using standard virus preparation or nucleic acid-based target templates. A correlation between real-time quantitative PCR and the conventional PCR for measles virus is highly desirable. PMID- 12629661 TI - Effect of immune modulation on TT virus (TTV) and TTV-like-mini-virus (TLMV) viremia. AB - The present study was designed to investigate how two chronically replicating viruses, TT virus (TTV) and TTV-like mini virus (TLMV), interact with host defence systems. Successive serum samples from three groups of subjects, undergoing modifications of their antiviral defence, were tested by real-time PCR to measure changes in viral titers, and by sequence analyses to indicate whether increases in viremia could be attributed to infection with an unfamiliar strain: 1) in patients receiving immunosuppressants subsequent to kidney transplantation, viral titers tended to increase; 2) in soldiers undergoing extreme training known to cause immunosuppression, insignificant increases in titers were observed; and 3) interferon treatment of patients with hepatitis C virus caused a temporary decrease in TTV and TLMV titers. Increases in viremia were associated only occasionally with the appearance of novel strains. The above results add to knowledge on how these viruses are influenced by the host. PMID- 12629662 TI - False resurrections: distinguishing regenerated from spared axons in the injured central nervous system. AB - Several recent studies report that axon regeneration can be induced in the mature mammalian nervous system by novel treatments or genetic manipulations. In assessing these reports, it is important to be mindful of the history of regeneration research, which is littered with the corpses of studies that reported regeneration that later proved incorrect. One important reason is the "spared axon conundrum," in which axons that survive a lesion are mistakenly identified as having regenerated. Here, we illustrate the problem and propose criteria that may be used to identify regenerated vs. spared axons, focusing on the injured spinal cord. PMID- 12629663 TI - Neurotrophins and taste buds. PMID- 12629664 TI - Brain-derived neurotrophic factor is present in adult mouse taste cells with synapses. AB - Brain-derived neurotrophic factor (BDNF), one of the members of the nerve growth factor family of neurotrophins, is expressed in developing gustatory papillae and is thought to be the neurotrophin that supports gustatory innervation during development. BDNF expression does not cease after development but continues in some taste cells of adult mice. To determine which types of taste cells produce BDNF, we undertook an immunohistochemical study of taste cells in BDNF(LacZ) gene targeted "knock-in" adult mice. In these mice, beta-galactosidase (beta-gal) immunoreactivity is an indicator of cells that produce BDNF transcripts. In the tongues of adult BDNF(LacZ) mice, beta-gal (BDNF) is present in long slender taste cells, as well as pyriform taste cells. Bromodeoxyuridine labeling experiments in BDNF(LacZ) mice indicate that BDNF is not present in taste cells that are younger than 3 days postmitotic. BDNF mainly colocalizes with markers of type II and type III taste cells: ubiquitin carboxyl terminal hydrolase (PGP 9.5), serotonin (5-HT), neural cell adhesion molecule (N-CAM), synaptic associated protein of 25 kDa (SNAP-25), and to a lesser extent with alpha gustducin. beta-Gal immunoreactivity is not associated with blood group H antigen, a marker of type I taste cells. We conclude that BDNF is absent from basal cells and type I (blood group H antigen immunoreactive) taste cells but is present in differentiated type II and type III taste cells. The presence of SNAP 25 in BDNF-expressing cells suggests a role for BDNF in synaptic formation and transmission. PMID- 12629665 TI - Heterogeneous expression of SNAP-25 and synaptic vesicle proteins by central and peripheral inputs to sympathetic neurons. AB - Neurons in prevertebral sympathetic ganglia receive convergent synaptic inputs from peripheral enteric neurons in addition to inputs from spinal preganglionic neurons. Although all inputs are functionally cholinergic, inputs from these two sources have distinctive neurochemical and functional profiles. We used multiple labeling immunofluorescence, quantitative confocal microscopy, ultrastructural immunocytochemistry, and intracellular electrophysiologic recordings to examine whether populations of inputs to the guinea pig coeliac ganglion express different levels of synaptic proteins that could influence synaptic strength. Boutons of enteric intestinofugal inputs, identified by immunoreactivity to vasoactive intestinal peptide, showed considerable heterogeneity in their immunoreactivity to synaptosome-associated protein of 25 kDa (SNAP-25), synapsin, synaptophysin, choline acetyltransferase, and vesicular acetylcholine transporter. Mean levels of immunoreactivity to these proteins were significantly lower in terminals of intestinofugal inputs compared with terminals of spinal preganglionic inputs. Nevertheless, many boutons with undetectable levels of SNAP 25 immunoreactivity formed morphologically normal synapses with target neurons. Treatment with botulinum neurotoxin type A (20-50 nM for 2 hours in vitro) generated significant cleavage of SNAP-25 and produced similar dose- and time dependent inhibitions of synaptic transmission from all classes of inputs, regardless of their mean level of SNAP-25 expression. The simplest interpretation of these results is that only synaptic boutons with detectable levels of SNAP-25 immunoreactivity contribute significantly to fast cholinergic transmission. Consequently, the low synaptic strength of intestinofugal inputs to final motor neurons in sympathetic pathways may be due in part to the low proportion of their boutons that express SNAP-25 and other synaptic proteins. PMID- 12629666 TI - "Dormant basket cell" hypothesis revisited: relative vulnerabilities of dentate gyrus mossy cells and inhibitory interneurons after hippocampal status epilepticus in the rat. AB - The "dormant basket cell" hypothesis suggests that postinjury hippocampal network hyperexcitability results from the loss of vulnerable neurons that normally excite insult-resistant inhibitory basket cells. We have reexamined the experimental basis of this hypothesis in light of reports that excitatory hilar mossy cells are not consistently vulnerable and inhibitory basket cells are not consistently seizure resistant. Prolonged afferent stimulation that reliably evoked granule cell discharges always produced extensive hilar neuron degeneration and immediate granule cell disinhibition. Conversely, kainic acid induced status epilepticus in chronically implanted animals produced similarly extensive hilar cell loss and immediate granule cell disinhibition, but only when granule cells discharged continuously during status epilepticus. In both preparations, electron microscopy revealed degeneration of presynaptic terminals forming asymmetrical synapses in the mossy cell target zone, including some terminating on gamma-aminobutyric acid-immunoreactive elements, but no evidence of axosomatic or axoaxonic degeneration in the adjacent granule cell layer. Although parvalbumin immunocytochemistry and in situ hybridization revealed decreased staining, this apparently was due to altered parvalbumin expression rather than basket cell death, because substance P receptor-positive interneurons, some of which contained residual parvalbumin immunoreactivity, survived. These results confirm the inherent vulnerability of dendritically projecting hilar mossy cells and interneurons and the relative resistance of dentate inhibitory basket and chandelier cells that target granule cell somata. The variability of hippocampal cell loss after status epilepticus suggests that altered hippocampal structure and function cannot be assumed to cause the spontaneous seizures that develop in these animals and highlights the importance of confirming hippocampal pathology and pathophysiology in vivo in each case. PMID- 12629667 TI - Ultrastructural examination of the somatic innervation of ventrotubercular cells in the rat. AB - Ventrotubercular cells are multipolar cells in the ventral cochlear nucleus (VCN) that project a collateral axon to the ipsilateral dorsal cochlear nucleus (DCN). These cells are thought to be involved in sensitizing DCN output neurons to spectral shapes that represent the location of a sound source in space. The present report focused on the neuronal composition of this pathway. Intracellular labeling studies in cats and mice have described two types of ventrotubercular cells (Smith and Rhode [1989] J Comp Neurol. 282:595-626; Oertel et al. [1990] J Comp Neurol. 295:136-154). In cats, one difference between the two classes is that type I multipolar neurons have fewer than 35% of their somata apposed by terminals, whereas type II cells have greater than 70% apposition values. Intracellular recordings from single cells, however, are difficult and thus limit the yield of data. We investigated whether a two-component description of the ventrotubercular pathway was representative of a larger population. This issue was addressed by retrogradely labeling ventrotubercular neurons with an extracellular injection of biotinylated dextran amine into the DCN of rats. These injections labeled many VCN neurons, thus providing a more complete view of the pathway than previous studies. Thirty-eight labeled cells were selected for electron microscopic analysis with respect to their location, cell body size, and ultrastructural morphology. We observed labeled type I and type II neurons, but unlike ventrotubercular cells in cats, many of these neurons in rats (17 of 38 cells) had appositions between 35% and 70%. On the basis of this analysis, a third class of ventrotubercular cell, called the adendritic neuron, was revealed. Adendritic neurons have small somata with many filopodial appendages, no observable dendrites, and high percentage of terminal appositions (>80%). The results demonstrated that the ventrotubercular pathway in the rat is diverse. PMID- 12629668 TI - Chemical coding of the human gastrointestinal nervous system: cholinergic, VIPergic, and catecholaminergic phenotypes. AB - The aim of this investigation was to identify the proportional neurochemical codes of enteric neurons and to determine the specific terminal fields of chemically defined nerve fibers in all parts of the human gastrointestinal (GI) tract. For this purpose, antibodies against the vesicular monoamine transporters (VMAT1/2), the vesicular acetylcholine transporter (VAChT), tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DBH), serotonin (5-HT), vasoactive intestinal peptide (VIP), and protein gene product 9.5 (PGP 9.5) were used. For in situ hybridization (35)S-labeled VMAT1, VMAT2, and VAChT riboprobes were used. In all regions of the human GI tract, 50-70% of the neurons were cholinergic, as judged by staining for VAChT. The human gut unlike the rodent gut exhibits a cholinergic innervation, which is characterized by an extensive overlap with VIPergic innervation. Neurons containing VMAT2 constituted 14-20% of all intrinsic neurons in the upper GI tract, and there was an equal number of TH-positive neurons. In contrast, DBH was absent from intrinsic neurons. Cholinergic and monoaminergic phenotypes proved to be completely distinct phenotypes. In conclusion, the chemical coding of human enteric neurons reveals some similarities with that of other mammalian species, but also significant differences. VIP is a cholinergic cotransmitter in the intrinsic innervation of the human gut. The substantial overlap between VMAT2 and TH in enteric neurons indicates that the intrinsic catecholaminergic innervation is a stable component of the human GI tract throughout life. The absence of DBH from intrinsic catecholaminergic neurons indicates that these neurons have a dopaminergic phenotype. PMID- 12629669 TI - New findings on cortical anatomy and implications for investigating the evolution of language. PMID- 12629670 TI - Variability of Broca's area homologue in African great apes: implications for language evolution. AB - The cortical circuits subserving neural processing of human language are localized to the inferior frontal operculum and the posterior perisylvian region. Functional language dominance has been related to anatomical asymmetry of Broca's area and the planum temporale. The evolutionary history of these asymmetric patterns, however, remains obscure. Although testing of hypotheses about the evolution of language areas requires comparison to homologous regions in the brains of our closest living relatives, the great apes, to date little is known about normal interindividual variation of these regions in this group. Here we focus on Brodmann's area 44 in African great apes (Pan troglodytes and Gorilla gorilla). This area corresponds to the pars opercularis of the inferior frontal gyrus (IFG), and has been shown to exhibit both gross and cytoarchitectural asymmetries in humans. We calculated frequencies of sulcal variations and mapped the distribution of cytoarchitectural area 44 to determine whether its boundaries occurred at consistent macrostructural landmarks. A considerable amount of variation was found in the distribution of the inferior frontal sulci among great ape brains. The inferior precentral sulcus in particular was often bifurcated, which made it impossible to determine the posterior boundary of the pars opercularis. In addition, the distribution of Brodmann's area 44 showed very little correspondence to surface anatomy. We conclude that gross morphologic patterns do not offer substantive landmarks for the measurement of Brodmann's area 44 in great apes. Whether or not Broca's area homologue of great apes exhibits humanlike asymmetry can only be resolved through further analyses of microstructural components. PMID- 12629671 TI - Trabecular rod thickness by direct measurement from 3D SEM anaglyphs. AB - This study presents a methodology for measuring the thickness of trabecular rods directly from anaglyphs. Macerated sagittal slices of T12 vertebral bodies from 15 subjects were examined by scanning electron microscopy (SEM). Two digital images (the second image tilted 5 degrees ) were recorded, and a 3D anaglyph was created. The thickness of the trabecular rods (Tb.Th((rods))), and the anatomical orientation of the trabecular rods were measured using an image analyser. Conventional 2D histomorphometry was performed on adjacent bone slices. A total of 1559 rod measurements were made from the 15 vertebral bone slices, with a mean Tb.Th((rods)) of 123 +/- 36 microm. The rod thickness in males (128 +/- 34 microm) was significantly greater than that in females (119 +/- 37 microm, P < 0.001). Tb.Th((rods)) changed significantly with age in the males: the thicker rods in the younger men reduced with age to a thickness similar to that in women. 3D measurements were significantly larger than the 2D estimates, and there was no correlation between the two methods of measurement. An inverse correlation was found between the number of rods and the bone volume fraction (BV/TV), indicating that decreased BV/TV is associated with an increased number of rods. The vertical rods (132 +/- 39 microm) were significantly thicker than the horizontal rods (116 +/- 33 microm, P < 0.001). The determination of rod numbers, and their orientation and individual thicknesses enables a greater understanding of cancellous bone architecture in both individuals and populations, and will allow more reliable finite element modelling. Direct measurements from 3D anaglyphs of intact specimens provide new data that show previously unrecognised age- and sex related changes. PMID- 12629672 TI - Frontonasal dysplasia in 3H1 Br/Br mice. AB - The adult Brachyrrhine (3H1 Br/+) mouse displays severe midfacial retrognathia, with a "pugnose" external appearance, but information concerning craniofacial morphology of the homozygote (3H1 Br/Br) mutant is lacking. This study characterized craniofacial phenotype and genotypic features of the homozygous condition. Segregation analysis was performed by phenotypic scoring of offspring from 3H1 Br/+ reciprocal matings. Whole-mount staining was undertaken to determine the presence or absence of cranial base structures in newborn and adult mice, while features of cranial base chondrification were examined using light microscopy and type II collagen immunohistochemistry. Karyotype analysis was performed to determine whether gross chromosomal aberrations were present. Finally, microsatellite mapping analysis was undertaken to provide further resolution of the Br locus. Results showed that Br was inherited as an autosomal semidominant feature. 3H1 Br/Br mice consistently lacked a presphenoid (with its lateral projections, including a preoptic root, postoptic root, and lesser wing). Karyotyping did not reveal major gross aberrations; however, microsatellite analysis localized Br to distal mouse chromosome 17 in the vicinity of D17Mit155. These results indicated that 3H1 Br/Br mice show characteristic features of frontonasal dysplasia, including median facial clefting and bifid cranium, as well sphenoidal malformations. Furthermore, this mutant should serve as a useful model for examining mechanisms of frontonasal dysplasia. PMID- 12629673 TI - Recruitment of intra- and extracardiac cells into the myocardial lineage during mouse development. AB - The tubular heart differentiates from the bilateral cardiac fields in the splanchnic mesoderm. The expression of smooth muscle proteins has been shown to accompany the early phases of cardiac muscle formation. In this study we show that during elongation of the arterial pole of the mouse linear heart tube, alpha smooth muscle actin (alpha-Sma) expression extends in the area that has been shown to become recruited into the myocardial lineage, but does not yet express myocardial markers. These data suggest that alpha-Sma identifies mesodermal cells that during subsequent development will be recruited into the myocardial lineage. Myocardium formation is not only observed at the arterial pole, but also at the venous pole and in the intracardiac mesenchyme. This results in the formation of the caval and pulmonary myocardium, the smooth-walled atrial myocardium, the myocardial atrioventricular septum, and the myocardial outlet septum. To determine whether recruitment into the myocardial lineage also takes place in these regions, the spatiotemporal pattern of expression of alpha-Sma and of the myocardial markers sarcoplasmatic reticulum calcium ATPase (Serca2a), alpha myosin heavy chain (Mhc), and beta-Mhc were examined. We show that prior to the expression of myocardial markers, alpha-Sma is expressed in these regions, which suggests that these mesodermal cells become recruited into the cardiac lineage after formation of the linear heart tube. PMID- 12629674 TI - Inhibition of the cardiac alpha-actin gene in embryonic cardiac myocytes by dominant-negative serum response factor. AB - Serum response factor (SRF), a transcription factor ubiquitously involved in the processes of cellular proliferation and differentiation, has been implicated in cardiac and skeletal muscle development because of its strong expression in embryonic muscle lineages, and its necessity for the transcription of transiently transfected muscle genes that contain SRF binding sites. This study was designed to ascertain whether SRF is required for the expression of an endogenous SRF dependent gene during differentiation of early embryonic cardiac myocytes by introducing a dominant-negative SRF construct via retroviral delivery. Although no effect on overt cellular differentiation was detected, semi-quantitative RT PCR revealed that expression of the SRF-dependent gene cardiac alpha-actin was inhibited, whereas expression of the non-SRF-dependent genes glyceraldehyde-3 phosphate dehydrogenase (GAPDH) and cardiac troponin-C was unaffected. No effect on myocyte proliferation was detected. Curiously, immunohistochemical localization of SRF protein suggested that whereas endogenous SRF was homogeneously dispersed throughout the cytoplasm and nucleus, the dominant negative SRF protein was concentrated in the nucleus. These results extend previous findings using transiently transfected genes to the endogenous level, indicating that SRF is required for the full expression of muscle genes that contain SRF binding sites during cardiac myocyte differentiation. PMID- 12629675 TI - Ectodysplasin receptor-mediated signaling is essential for embryonic submandibular salivary gland development. AB - Hypohidrotic (anhidrotic) ectodermal dysplasia (HED), the most common of the approximately 150 described ectodermal dysplasias, is a disorder characterized by abnormal hair, teeth, sweat glands, and salivary glands. Mutations in the EDA (ectodysplasin-A) and EDAR (ectodysplasin-A receptor) genes are responsible for X linked and autosomal HED, respectively. Abnormal phenotypes similar to HED are seen in Tabby (Eda(Ta)) and downless (Edar(dl)) mutant mice. Although recent studies have focused on the role of Eda/Edar signaling during hair and tooth development, very little is known about its role during embryonic submandibular salivary gland (SMG) development. To this end, we analyzed the SMG phenotypes in Tabby (Ta) and downless (dl) mutant mice and determined that Ta SMGs are hypoplastic, whereas dl SMGs are severely dysplastic. The absence of SMG ducts and acini in dl SMGs suggests that Eda/Edar signaling is essential for lumina formation and glandular histodifferentiation. Our localization of Eda and Edar proteins at sites of lumen and acini formation supports this conclusion. Moreover, the presence of SMGs in both Ta and dl mutant mice, as well as the absence of immunodetectable Eda and Edar protein in Initial Bud and Early Pseudoglandular stage SMGs, indicate that Eda/Edar-mediated signaling is important for branching morphogenesis and histodifferentiation, but not for initial gland formation. To initially delineate the morphoregulatory role of Eda/Edar-mediated signaling during embryonic SMG development, we cultured embryonic day 14 SMGs with enhanced or abrogated Eda/Edar signaling. Eda supplementation induced a significant increase in SMG branching, and enhanced activation of NF-kappaB. Abrogating Eda/Edar signaling by adding the soluble form of Edar to bind endogenous ligand in embryonic SMGs results in a significant dose dependent decrease in branching morphogenesis. Taken together, our results suggest that the Eda/Edar/NF-kappaB pathway exerts its effect on SMG epithelial cell proliferation, lumina formation, and histodifferentiation. PMID- 12629676 TI - Ultrastructural morphometry of capillary basement membrane thickness in normal and transgenic diabetic mice. AB - Capillary basement membrane (CBM) thickening is an ultrastructural hallmark in diabetic patients and in animal models of diabetes. However, the wide variety of tissues sampled and diverse methods employed have made the interpretation of thickness data difficult. We showed previously that acellular glomerular BMs in OVE26 transgenic diabetic mice were thickened beyond normal age-related thickening, and in the current study we hypothesized that other microvascular BMs likewise would show increased widths relative to age-matched controls. Accordingly, a series of tissues, including skeletal and cardiac muscle, ocular retina and choriod, peripheral nerve, lung, pancreas, and renal glomerulus was collected from 300-350-day-old normal and transgenic mice. Transmission electron micrographs of cross sections through capillary walls were prepared, and CBM thickness (CBMT) was determined by the "orthogonal intercept" method. Morphometric analyses showed highly variable transgene-related BMT increases in the sampled tissues, with glomerular BM showing by far the greatest increase (+87%). Significant thickness increases were also seen in the retina, pulmonary alveolus, and thoracoabdominal diaphragm. BMT increases were not universal; however, most were modestly widened, and those that were thickest in controls generally showed the greatest increase. Although the pathogenesis of diabetes related increases in CBM is poorly understood, data in the current study showed that in OVE26 transgenic mice increased BMT was a frequent concomitant of hyperglycemia. Accordingly, it seems likely that hyperglycemia-induced microvascular damage may be a contributing factor in diabetic BM disease, and that microvessel cellular and extracellular heterogeneity may limit the extent of CBM thickening in diverse tissues. PMID- 12629677 TI - Ultrastructure indicative of ion transport in tectal, Deiters, and tunnel cells: differences between gerbil and chinchilla basal and apical cochlea. AB - Ultrastructural examination revealed an epithelium of about five tectal cells (TCs) roofing the outer tunnel (OT) in the mid to upper, but not the basal, region of gerbil and chinchilla cochlea. Structures in TCs that are apparently specialized for retrieval of K(+) released into tunnel fluid from outer hair cells (OHCs) include surface fimbriae in the gerbil and canalicular reticulum in the chinchilla. A tunnel roof of organelle-rich TCs appeared to be better equipped for ion resorption than a roof composed of organelle-poor Hensen cells (HCs). Fimbriae, filopodia, and the cell body of TCs descended to contact the third Deiters cell (DC3) in the gerbil, and the hypertrophied DC3 phalanx rose to contact TCs in the chinchilla, which suggests a solute exchange between TCs and DCs. Previously unrecognized structures that are speculated to provide ATP ligand for cochlear purinoreceptors occurred in the chinchilla DC and gerbil TC. The observation of a microtubule stalk in DCs indicated that they also function in cochlear mechanics. A newly delineated lateral tunnel cell (LTC) intervened between the DC3 and HC in both species. The apicomedial plasmalemma of all DCs fitted closely to the base of OHCs and enveloped afferent nerves. The morphologic specializations reported here provide further support for the proposed transcellular lateral flow route for K(+) currents generated by sound exposure and neural activity. The previously demonstrated expansion of Boettcher cells, outer sulcus cell roots, type Il and IV fibrocytes, and apical microvilli on HCs and Claudius cells (CCs) in the base of the cochlea is postulated here to mediate a basal parallel current that could supply the increased K(+) transport required for the basally elevated electric potential (EP). PMID- 12629678 TI - Connexin43 is expressed in mouse fetal ovary. AB - Developmental studies have shown that connexin43 (Cx43) is expressed in the ovary from the first day of life and throughout the rest of postnatal development. In both mouse embryonic ovaries and testes, target-directed deletion of Cx43 gene induces a significant decrease in germinal cells, but the exact mechanism determining this reduction remains unknown. Moreover, recently we found that Cx43 is abundantly expressed in mouse testes from the earliest stages of its fetal development. In the present work we investigate whether Cx43 transcript and protein are expressed in mouse embryonic ovaries. Total RNA was analyzed with specific Cx43 oligonucleotides in RT-PCR studies. A Cx43 PCR product was detected in ovaries at 16.5 and 18.5 days postcoitum (dpc). Bands of 43-45 kDa, characteristic of Cx43, were detected in immunoblots of total homogenates of ovaries at 14.5 and 18.5 dpc. Cell type-specific expression of Cx43 was investigated using double-labeled sections incubated with specific antibodies against Cx43 and the enzyme 3beta-hydroxysteroid dehydrogenase (3betaHSD) or a germ cell nuclear antigen (GCNA1), which are cell markers of steroidogenic and germinal cells, respectively. At 18.5 dpc, Cx43 was found in conglomerates of 3betaHSD-positive cells. Cx43 was also localized at homocellular junctions between parenchyma pregranulosa cells, and at heterocellular junctions between pregranulosa and germinal cells. At these two latter localizations, Cx43 was traced back to 12.5 dpc. In conclusion, this study demonstrates for the first time that from the earliest stages of embryonic ovary development, Cx43 is expressed in principal cell types involved in control of female fertility. These data suggest that the gap junctions formed with Cx43 between somatic and germinal cells may be necessary for prenatal expansion of germinal cells at initial stages of fetal gonadal development. PMID- 12629679 TI - Serotonin in the rabbit ileum: localization, uptake, and effect on motility. AB - Repeated experiments to localise serotonin in the myenteric plexus of rabbit ileum failed. After preincubation in serotonin (10(-5) M), an extensive varicose fibre system was detected by immunocytochemical methods. Stained fibres left the myenteric plexus and ran to the muscle layers. Labelled cell bodies could not be found, even after pretreatment with colchicine or pargyline. Application of reserpine (10(-5) M) and fluoxetine (10(-5) M) prevented serotonin uptake. Antisera against tryptophan hydroxylase revealed a rich fibre system, including those processes that entered the tertiary plexus. These fibres were able to accumulate serotonin, but again the cell bodies could not be detected. Serotonin caused concentration-dependent contraction in the longitudinal muscle layer of the rabbit ileum. Pretreatment with tetrodotoxin strongly reduced the effect of serotonin. Preapplication of atropine caused a slight decrease of response evoked by serotonin. Combined administration of tetrodotoxin and atropine significantly reduced the responses to serotonin, but did not abolish them. At the same time, agonists of 5-HT(2) and 5-HT(4) receptors caused concentration-dependent contractions. Our studies show that: 1). Without pretreatment, serotonin cannot be detected in the myenteric plexus of rabbit ileum. 2). An extensive uptake system works in this plexus. If released from myenteric nerve fibres, serotonin may evoke contractions in indirect and direct ways. 3). There may be an extrinsic serotoninergic innervation from the mesenteric ganglia. 4). Serotonin exerts its effect through 5-HT(2) and 5-HT(4) receptors on smooth muscle cells and nerve elements. PMID- 12629680 TI - Disconnections. PMID- 12629681 TI - Construction and validation of a body image rating scale: a preliminary study. AB - PURPOSE: To construct and validate a preliminary version of a scale for use in routine screening of body image disturbances in nursing practice. METHODS: A five stage procedure consisting of item construction, analysis of content validity, analysis of reliability, analysis of construct validity, and analysis of the discriminant validity of the scale. FINDINGS: At the end of the current stage of development, the preliminary version of the scale consisted of 23 items, demonstrating stability, adequate internal consistency, and discriminant validity between subjects in the upper and lower groups of scores. CONCLUSIONS: Although the psychometric properties of the scale have been delineated, the scale needs further psychometric evaluation and testing in different settings/samples so its clinical validity and reliability can be confirmed. PRACTICE IMPLICATIONS: The preliminary version of the body image rating scale has the potential to provide the nurse with reliable and valid information about a person's mental image of his/her own body. Using it can help nurses to identify possible body image disturbances and to plan individual nursing interventions. PMID- 12629682 TI - Fatigued elderly patients with chronic heart failure: do patient reports and nurse recordings correspond? AB - PURPOSE: To compare descriptions of fatigue from interviews with elderly people with chronic congestive heart failure (CHF) and data recorded by nurses at an outpatient heart failure clinic. METHODS: Patients (N = 158) were screened for moderate to severe CHF and interviewed using a revised form of the Fatigue Interview Schedule (FIS). Seventy-nine of these patients were offered visits at a nurse-monitored heart failure clinic. Nursing documentation of fatigue in patient records (n = 56) at the heart failure clinic was compared to the patient interviews. FINDINGS: Results indicated poor concordance between patients' descriptions and record content concerning fatigue. Decreased libido was linked to fatigue according to the patients but not to the nurses' notes. Cognitive characteristics of fatigue were rarely recorded, but were more frequent in patient interviews. CONCLUSIONS: Nurses must recognize characteristics and factors related to fatigue in patients and develop strategies to help patients cope with their restricted ability in daily life. PRACTICE IMPLICATIONS: Using the words and expressions of the patients and the diagnostic characteristics of fatigue in record-keeping can support the nurses understanding of patients living with CHF. PMID- 12629684 TI - [Biosynthetic pathways for purine nucleotides and uric acid]. PMID- 12629683 TI - The intervention of surveillance across classification systems. AB - PURPOSE: To analyze the intervention of surveillance across nursing classification systems. DATA SOURCES: Published literature and book chapters. DATA SYNTHESIS: Surveillance is a clearly established nursing intervention used in multiple practice settings and in response to many diagnoses. The literature and definitions are divided into surveillance of populations and surveillance of individuals. CONCLUSIONS: General agreement exists on the process of surveillance, but not on whether there is a time element to surveillance. PRACTICE IMPLICATIONS: Research is needed on outcomes of surveillance and best practices established. The similarity in definitions of surveillance may allow comparison of outcomes when the diagnosis and practice setting are the same, regardless of the classification systems. PMID- 12629685 TI - [Phosphoribosylpyrophosphate(PRPP) synthetase]. PMID- 12629686 TI - [Amidophosphoribosyltransferase]. PMID- 12629687 TI - [GAR synthetase . GAR transformylase . AIR synthetase]. PMID- 12629688 TI - [FGAM synthetase]. PMID- 12629689 TI - [AIR carboxylase . SAICAR synthetase]. PMID- 12629690 TI - [The second half of de novo synthetic pathway of IMP and conversion of IMP to AMP]. PMID- 12629691 TI - [IMP dehydrogenase]. PMID- 12629693 TI - [Hypoxanthine guanine phosphoribosyltransferase (HPRT)]. PMID- 12629692 TI - [GMP synthetase]. PMID- 12629694 TI - [Adenine phosphoribosyltransferase (APRT)]. PMID- 12629695 TI - [5'-nucleotidase]. PMID- 12629696 TI - [Purine nucleoside phosphorylase (PNP)]. PMID- 12629697 TI - [Adenosine deaminase]. PMID- 12629698 TI - [Guanine deaminase]. PMID- 12629700 TI - [AMP deaminase (AMPD)]. PMID- 12629699 TI - [Xanthine dehydrogenase (xanthine oxidase)]. PMID- 12629702 TI - [Mode of existence and dynamics of uric acid in blood]. PMID- 12629701 TI - [GMP reductase]. PMID- 12629703 TI - [Urate transport in human kidney]. PMID- 12629704 TI - [Urate transport mechanism of mammalian cell membrane]. PMID- 12629705 TI - [Influence of sex and age on serum uric acid level]. PMID- 12629706 TI - [Effect of foods on serum uric acid level]. PMID- 12629707 TI - [Influence of exercise on serum uric acid]. PMID- 12629708 TI - [Effect of ethanol on uric acid level]. PMID- 12629709 TI - [Influence of pregnancy on serum uric acid]. PMID- 12629710 TI - [Definition and determination of serum uric acid level]. PMID- 12629711 TI - [Hyperuricemia--definition and classification]. PMID- 12629712 TI - [Typing of hyperuricemia (uric acid clearance)]. PMID- 12629713 TI - [Idiopathic hyperuricemia]. PMID- 12629714 TI - [Epidemiology of hyperuricemia]. PMID- 12629715 TI - [Diet therapy for management of hyperuricemia and gout]. PMID- 12629716 TI - [Exercise therapy for hyperuricemia]. PMID- 12629717 TI - [Anti hyperuricemic agents]. PMID- 12629718 TI - [Drug therapy for idiopathic hyperuricemia--introduction, dose, and side effects]. PMID- 12629719 TI - [Development antihyperuricemic candidates]. PMID- 12629720 TI - [Gouty arthritis]. PMID- 12629721 TI - [Tophaceous gout]. PMID- 12629722 TI - [Hyperuricemia and urolithiasis]. PMID- 12629723 TI - [Gouty kidney]. PMID- 12629724 TI - [Essential hypertension]. PMID- 12629725 TI - [Hyperlipidemia and obesity]. PMID- 12629727 TI - [Cardiovascular disease]. PMID- 12629726 TI - [Effect of hyperglycemia and insulin resistance on serum uric acid]. PMID- 12629728 TI - [Hyperuricemia and atherosclerosis]. PMID- 12629729 TI - [Sleep apnea syndrome]. PMID- 12629730 TI - [Familial juvenile hyperuricemic nephropathy]. PMID- 12629731 TI - [Hyperuricemia associated with inborn errors of purine metabolism: screening, enzymatic and genetic diagnosis]. PMID- 12629732 TI - [PRPP synthetase superactivity]. PMID- 12629733 TI - [Complete and partial deficiencies of hypoxanthine guanine phosphoribosyltransferase (HPRT)]. PMID- 12629734 TI - [Hyperuricemia due to renal failure, autosomal dominant polycystic kidney disease and lead nephropathy]. PMID- 12629736 TI - [Hyperuricemia in endocrine disorders]. PMID- 12629735 TI - [Secondary hyperuricemia in hematologic disease]. PMID- 12629737 TI - [Secondary hyperuricemia in glycogen storage disease types I, III, V and VII]. PMID- 12629738 TI - [Secondary hyperuricemia in burn and trauma]. PMID- 12629739 TI - [Secondary hyperuricemia in Down's syndrome]. PMID- 12629740 TI - [Hyperuricemia in preeclampsia]. PMID- 12629741 TI - [Idiopathic retroperitoneal fibrosis and hyperuricemia]. PMID- 12629742 TI - [Drug-induced hyperuricemia]. PMID- 12629743 TI - [Non-drug substance (over intake of foods which are contained purine body, fat and fructose)]. PMID- 12629744 TI - [Alcohol-related tissue injury and hyperuricemia]. PMID- 12629746 TI - [Typing of hypouricemia (uric acid clearance)]. PMID- 12629745 TI - [Hypouricemia--definition and classification]. PMID- 12629747 TI - [Idiopathic renal hypouricemia]. PMID- 12629748 TI - [Primary underproductive hypouricemia]. PMID- 12629749 TI - [Hereditary renal hypouricemia]. PMID- 12629750 TI - [PNP (purine nucleoside phosphorylase) deficiency]. PMID- 12629751 TI - [Hereditary xanthinuria and molybdenum cofactor deficiency]. PMID- 12629752 TI - [PRPP synthetase deficiency]. PMID- 12629753 TI - [Hypouricemia due to renal tubular dysfunction]. PMID- 12629754 TI - [Hypouricemia in diabetes mellitus]. PMID- 12629755 TI - [Secondary hypouricemia in patients with malignant disease]. PMID- 12629756 TI - [Hypouricemia associated with various liver diseases]. PMID- 12629757 TI - [Drug-induced hypouricemia: pathophysiology and application to the hyperuricemia with life style related disease]. PMID- 12629758 TI - [Pseudogout--calcium pyrophosphate dihydrate crystal deposition disease]. PMID- 12629759 TI - [Basic calcium phosphate crystal deposition disease]. PMID- 12629760 TI - [Rheumatoid arthritis]. PMID- 12629761 TI - [Osteoarthritis]. PMID- 12629762 TI - [Gouty attack and resembled diseases in forefoot]. PMID- 12629763 TI - [Introduction of the new guideline for the management of hyperuricemia and gout with special reference to its policy]. PMID- 12629765 TI - [Interleukin 6 deduces serum urate concentration]. PMID- 12629764 TI - [Molecular mechanism for urinary excretion of hypoxanthine and xanthine]. PMID- 12629766 TI - [Association of serum uric acid level with cardiovascular mortality]. PMID- 12629767 TI - [Blood antioxidant capacity in gout]. PMID- 12629768 TI - [Toward individualized medicine of gout]. PMID- 12629769 TI - [Gene therapy for purine-pyrimidine metabolism inborn errors]. PMID- 12629770 TI - [Animal models for abnormal purine metabolism]. PMID- 12629771 TI - [Contents of purine bases in foods and alcoholic beverages]. PMID- 12629772 TI - [Analysis of patients with septic shock due to urosepsis brought on by ureteral calculi]. AB - While a ureteral stone is a common disease, it occasionally causes urosepsis and septic shock. We analyzed 6 cases of septic shock due to urosepsis caused by a ureteral stone from August 1998 to September 2001. All patients were female, ranging in age from 38-76 years old (63.8 +/- 15.3). Stones ranged from 4 to 12 ml in size. The results of analysis of bacterial culture from blood and urine revealed E. coli in 4 cases, K. pneumoniae in 1 case and P. mirabilis in 1 case. Treatment, including intravenous transfusion, the administration of vasopressor drugs and antibiotics, and anti-disseminated intravascular coagulation (DIC) treatments were performed. Percutaneous nephrostomy in 4 cases and ureteral stent indwelling in 1 case were also used for the management of urinary tract. Extracorporeal shock wave lithotripsy (ESWL) was performed in 4 patients after improvement of general condition. Furthermore, the 3 most recent patients had also received therapy to remove endotoxins. All patients were diagnosed in a stone-free condition after a combination of these therapies. The mean recovery interval from shock condition to shock-free status seemed to be shorter in the patients that received the endotoxin removal therapy. PMID- 12629773 TI - The significance of prostate-specific antigen alpha-1-antichymotrypsin complex and its indices for the detection of prostate cancer. AB - We examined the usefulness of prostate specific antigen alph-1-antichymotrypsin complex (PSA-ACT) and its indices for the detection of prostate cancer in patients with a prostate specific antigen (PSA) level between 2.1 and 10.0 ng/ml. Between July 1999 and October 2001, 151 patients with a PSA level between 2.1 and 10.0 ng/ml underwent a systematic biopsy under transrectal ultrasound (TRUS) guidance. The clinical values of total PSA, PSA-ACT, PSA density (PSAD), PSA-ACT density (PSA-ACTD), PSA transition zone density (PSATZD) and PSA-ACT transition zone density (PSA-ACTTZD) for the detection of prostate cancer were compared by using receiver operating characteristic (ROC) curve analysis. Of the 151 patients, 36 (23.8%) were histologically confirmed as having prostate cancer. The differences between patients with prostate cancer and benign prostatic disease were significant with respect to the PSA and PSA-ACT related parameters examined in this study. According to ROC curve analysis, the area under the curve (AUC) of PSA-ACTTZD was the greatest of all the parameters. The differences was significant between the AUC of PSA-ACTTZD and total PSA (p < 0.05). The cutoff value of PSA-ACTTZD with 0.20 ng/ml2 showed the highest sum of sensivitity (90%) and specificity (55%). Also, in 86 patients with a PSA level between 2.1 and 6.0 ng/ml, the AUC of PSA-ACTTZD was the greatest of all the parameters. Measuring the level of PSA-ACT and its indices may provide a better differentiation of prostate cancer and benign prostatic disease than total PSA alone in patients with intermediate PSA levels. PSA-ACTTZD is the most useful indicator among PSA ACT and its volume indices. PMID- 12629774 TI - Clinical comparative evaluation of radical retropubic and perineal prostatectomy approaches for prostate cancer. AB - We compared the outcomes, advantages, and disadvantages of retropubic and perineal approaches to radical prostatectomy for prostate cancer. From 1990 to 2000, 37 patients (average age: 66.6 years) who underwent radical retropubic prostatectomy (RRP) and 30 patients (average age: 70.1 years) who underwent radical perineal prostatectomy (RPP) were identified from the database of our department. Outcome measures included operative duration, estimated blood loss, blood transfusions, positive margins, complications, and incontinence rates. There were no differences between the RRP and RPP groups in complications, incontinence rates, or positive margins. The mean operative duration was 228 min in the RRP group and 198 mm in the RPP group (p < 0.05). The mean estimated blood loss was 1,060 ml in the RRP group and 717 ml in the RPP group (p < 0.01). The mean volume of blood transfusions was 620 ml in 17 patients in the RPP group and 700 ml in one patient in the RPP group (p < 0.001). In conclusion, the clinical results of RRP and RPP groups were similar; the advantages of the perineal approach were shorter operative duration, smaller estimated blood loss and less blood transfusion. PMID- 12629775 TI - [Marked improvement of cardiac function following renal transplantation: a case report]. AB - The recipient, a 31-year-old man, was diagnosed with severe diffuse hypokinesis of left ventricle and diastolic dysfunction by echocardiography before the renal transplantation. His left ventricular ejection fraction (LVEF) was 29%. Therefore we had to postpone the operation and tried to improve his cardiac function. We controlled his dry weight more strictly with hemodialysis and limited in take of salt and water to reduce preload of his heart and administered erythropoietin to improve his anemic condition. We administered angiotensin-converting enzyme inhibitors (ACEI) to control his blood pressure and reduce afterload of his heart. Two months later, his cardiac function was marked improved (LVEF, 29% to 47%), and he underwent renal transplantation. Five months after the operation, improvement of his cardiac function kept on being observed in spite of the cessation of treatment for his heart (LVEF, 47% to 68%). Successful renal transplantation can significantly improve the cardiac function of patients on maintenance dialysis. PMID- 12629776 TI - [Papillary adenocarcinoma of the prostate: report of 4 cases]. AB - Papillary adenocarcinoma of the prostate, previously referred to as endometrioid carcinoma, is a variant of prostatic adenocarcinoma. Clinical and pathological evidence of involvement of the periurethral prostatic duct or verumontanum is usually required for definitive diagnosis of papillary adenocarcinoma. However, significant histologic and clinical features of papillary adenocarcinoma overlap with typical acinar carcinoma. Four cases of papillary adenocarcinoma were studied for the clinical features, histologic characteristics and immunohistochemical nature of prostatic specific antigen. In two cases, there were papillary regions, near the verumontanum, but in the other two cases, there were no papillary regions in the urethra. In two cases, acinar adenocarcinoma coexisted with papillary adenocarcinoma. All cases displayed positive immunohistochemical staining for prostatic specific antigen. In accordance with the observations of others, we suggest that papillary adenocarcinoma is one aspect of growth pattern of acinar adenocarcinoma, not a concept of a unique clinical and pathological entity. PMID- 12629777 TI - [A case of psoas abscess with ureteral stone]. AB - A 62-year-old diabetic woman with right hydronephrosis was referred to our department, presenting right back pain and high fever. A ureteral stone with right hydronephrosis was pointed on DIP. Although the calculus was removed by extracorporeal shock wave lithotripsy, persistent high fever continued. Abdominal computed tomography (CT) and magnetic resonance imaging (MRI) revealed a mass in the retroperitoneal space. Purulent fluid with a negative cytology was acquired by percutaneous biopsy following ultrasonography-guided percutaneous drainage. The postoperative course was uneventful with antibiotic therapy. PMID- 12629778 TI - [Fibroepithelial polyp of the anterior urethra, found during post-operative follow-up of ureteral cancer]. AB - A 52-year-old man underwent nephroureterectomy against left distal ureteral cancer (transitional cell carcinoma, Grade 2, pTa), and was followed up by surveillance alone. Twenty-two months postoperatively, a pedunculated tumor was found in the anterior urethra by cystourethroscopy. Pathological examination of the tumor resected transurethrally revealed a fibroepithelial polyp. There have been only twelve cases of fibroepithelial polyp of the anterior urethra reported in the literature. Since 4 out of 8 adult cases among them were found during the postoperative follow up period of urothelial cancer, mechanical irritation by repeated transurethral procedures might be responsible for the development of fibroepithelial polyp of the anterior urethra in these cases. If anterior urethral tumor is found during follow-up urothelial carcinoma, benign urethral polyp should be considered in the differential diagnosis. PMID- 12629779 TI - [Usefulness of nocturnal urethral indwelling catheter for treating bladder dysfunction with nocturnal polyuria: case report of 3 women]. AB - Clean intermittent catheterization is a well-known procedure of urinary drainage for patients who are unable to empty the bladder sufficiently. However, some patients with bladder dysfunction and nocturnal polyuria fail to obtain the benefits of intermittent catheterization and have annoying symptoms of nocturnal incontinence and low-compliance bladder, which threaten both their quality of life and renal function. We report the usefulness of nocturnal urethral indwelling catheterization using a specially designed catheter to treat patients (three women) with lower urinary tract dysfunction and nocturnal polyuria. Case 1: A 45-year-old woman with mental retardation suffered from difficulty of micturition and residual urine. A nocturnal urethral indwelling catheter freed her from difficulty with micturition and residual urine. Case 2: A 28-year-old woman with spina bifida and neuropathic bladder dysfunction suffered from urinary incontinence and recurrent pyelonephritis. The recurrent pyelonephritis was prevented and bladder compliance was improved with use of the nocturnal urethral indwelling catheter. Case 3: A 66-year-old woman with cervical myelopathy and multiple episodes of cerebral infarction suffered from nocturnal urinary incontinence. She underwent clean intermittent catheterization by her husband. Use of the nocturnal urethral indwelling catheter solved the problem of her nocturnal incontinence and relieved her husband of her nocturnal care. Nocturnal urethral indwelling catheterization is useful for treatment of nocturnal incontinence and recovery of bladder compliance in patients with lower urinary tract dysfunction and nocturnal polyuria. PMID- 12629780 TI - [Leydig cell tumor of the testis presenting male infertility: a case report]. AB - A 33-year-old male was referred to our hospital for male infertility with painless swelling of the left scrotal content. Left high orchiectomy was performed under the diagnosis of left testicular tumor. Histologically, this testicular mass was a Leydig cell tumor. We reviewed 55 cases of Leydig cell tumor of the testis previously reported in Japan, and reported the hormonal profile in our case before and after surgery. PMID- 12629781 TI - [Superselective endovascular treatment of renal artery aneurysms with detachable microcoils]. AB - Selective transcatheter embolization using an interlocking detachable coil was performed in two cases with renal artery aneurysms. Case 1, a 73-year-old male, was referred to our department complaining of macrohematuria persisting for three days. Case 2, a 52-year-old female, visited our department because calcifications in both renal pedicle areas were found in a health check up. Drip infusion pyelography (DIP) and computerized tomography (CT) revealed in case 1 an aneurysm which was 11 x 10 cm in diameter with nearly circumferential calcification in the right renal pedicle area and in case 2 two aneurysms, one 12 x 10 cm in the right and one 22 x 20 cm in the left with imperfect calcification. Enhanced CT confirmed blood flow in the aneurysms. Because abdominal surgery, was considered to be a highrisks in both cases 1 and 2 the coil embolization was performed. The patients have been followed up after the embolization without any particular complications or changes in the coil location. PMID- 12629782 TI - [Diagnostic availability of magnetic resonance angiography (MRA) in a case of cirsoid-type renal arteriovenous fistula]. AB - We report a case of cirsoid-type renal arteriovenous fistula (RAVF), which was difficult to differentiate from renal pelvic tumor by intravenous pyelography and computed tomographic (CT) scan. A 76-year-old woman visited our department complaining of sudden asymptomatic macroscopic hematuria. A CT scan showed an irregular mass (3 x 2 x 2 cm) in the right kidney. MRA revealed a cirsoid-type renal arteriovenous fistula. The patient was treated with TAE using ethanol, geratin sponge, and a coil. It is suggested that MRA is useful for the diagnosis of renal arteriovenous fistula. PMID- 12629784 TI - The 2001 IAHSS survey--crime in hospitals. AB - The 2001 IAHSS crime survey involved the participation of more hospitals--295- than in any of the past ten years. The lower levels of Larceny Theft revealed in prior surveys continued in 2001. In five of the seven categories of violent crime, however, per hospital increases in incidents were reported, with sharp increases in Robbery and Aggravated Assaults. PMID- 12629783 TI - [Transcatheter arterial embolization with n-butyl 2-cyanoacrylate (hystoacryl) for renal arteriovenous malformation: case report]. AB - A 22-year-old woman presented with sudden onset gross hematuria. Drip infusion pyelography and enhanced computerized tomography yielded no unusual findings. Renal angiography demonstrated an arteriovenous malformation (AVM) in the central portion of the right kidney. Superselective transcatheter arterial embolization (TAE) of the AVM was performed with a steel coil, and hematuria disappeared after TAE. However, severe gross hematuria developed again six days later. Second renal arteriography revealed the formation of the collateral vessels. By using n-butyl 2-cyanoacrylate (NBCA), TAE for AVM was performed successfully. She has been free of hematuria during the one-year follow-up. PMID- 12629785 TI - 15 essential leadership skills for healthcare security managers. AB - How are healthcare security managers managing their departments? Some 15 leadership skills essential to healthcare management are described. PMID- 12629786 TI - Are your disaster plans ready, really ready? AB - When a real disaster takes place, a hospital has a chance to really test the effectiveness of its disaster plan. Here is a detailed report on what happened when such an incident occurred. PMID- 12629787 TI - Developing a workable environment of care management plan. AB - A review of the basic points to be included in an EC plan for JCAHO based on the author's experiences in healthcare protection services. PMID- 12629788 TI - Psychiatric hospital challenges for healthcare security officers. AB - Security and Safety managers in today's healthcare facilities need to use creative thinking and resourcefulness, to juggle competing issues in psychiatric hospitals, wards, or units. Using a 3-step process of accountability, access control, and scenario exercises, these managers can mitigate the real-world risk assessment discoveries that might not be evident in well-documented facility policies, staff training, or even written surveys. PMID- 12629789 TI - Employee screening: issues for the healthcare industry. AB - Approaches and opportunities for healthcare management to meet increasing demands for better pre-employment background checking are presented in this analysis of the various factors involved. PMID- 12629790 TI - Detection of drug abuse by health professionals. AB - Analyzing prescription drugs in urine is a challenge for both the lab and the agency monitoring health professionals. PMID- 12629791 TI - Training for tense times. AB - To find out what companies are doing to address workplace violence, Security Management magazine interviewed security directors in different industries. Here is the report on the workplace violence response team at North Memorial Health Care, Robbinsdale, MN. PMID- 12629792 TI - Infant abduction: prevention through a multidisciplinary and multidimensional approach. AB - Everything that could or should be involved in preventing an infant abduction is reviewed in this detailed article. PMID- 12629793 TI - Closing the training gap in hospital security. AB - The need to adequately train both security and clinical staff members, as well as other employees, in security can only be met by having effective training programs in place. Health care security leaders explain what such training should consist of. PMID- 12629794 TI - Who stole my identity? AB - What you personally and as a manager should know about identity theft, its ramifications, and how to help your institution prevent potential thieves from stealing confidential information. PMID- 12629795 TI - Special programs for foreign patients and their security concerns. AB - Hospitals that admit many foreign patients have been revising their international programs since 9/11. PMID- 12629796 TI - Nurses learn vital signs of safety. AB - Protecting home health nurses and other professionals is a growing problem for healthcare security. The authors present their recommendations for an effective home healthcare safety program. PMID- 12629797 TI - [Experimental and histological analysis of histogenesis and tissue regeneration (some results of the 20th century and perspectives on further research)]. AB - The article is devoted to theoretical analysis of certain achievements in the area of cellular and tissue biology. Evolutional laws of tissue development, theory of divergent differentiation of histological structures, conception of cellular-differon tissue organization, formulated earlier were completely suitable for analysis of processes of development and regeneration of living systems at different structural levels of organization. System-forming plasmalemmas, receptor-transductor and adaptive properties were related to most important characteristics of living systems that can be experimentally histologically analysed. Morphofunctional signs of differons and histions and tissue classifications were considered. PMID- 12629798 TI - [Histological structure of adult mouse brain in protein-deficient nutrition]. AB - The effect of protein deficient nutrition on histostructure of different regions of brain of mature animals was studied in the experimental model of protein and energy deficiency. Protein deficient nutrition was shown to exert negative influence on structural and functional state of brain neurons which is manifested through increase of dystrophic changes in neurons of all brain regions, accumulation of lipofuscine in neurons and complex of structural changes in synaptic contacts. PMID- 12629799 TI - [Spatial organization of the corticopalladial projection system of dog brain]. AB - Spatial organization of corticopallidal projectional system was studied in 11 outbred dogs by method based on horse radish peroxidase transport. It was demonstrated that globus pallidum receives projections predominantly from neocortical zones (motor, premotor, somatosensory, parietal and auditory and from insular field of mesocortex. Mesocortical (prelimbic, orbital and insular) and allocortical (entorhinal, piriform and periamygdalar) including archicortex (subicular part of hippocampal formations) fields project onto ventral pallidum. Entopeduncular nucleus receives projections from neocortical zones (motor, premotor, somatosensory, parietal and auditory), mesocortex (prelimbic, orbital, insular and cingular fields) and allocortex (entorhinal and periamygdalar fields). The data obtained indicate specificity of distribution of cortical afferent projectional fibres in each of nuclei studied which allows to consider globes pallidum as motor zone and ventral pallidum as limbic zone of paladial complex. As projections from functionally different cortical fields were revealed in entopeduncular nucleus it may be suggested that this is the exact site for interaction of functionally different information, including the one received from the cortex. PMID- 12629800 TI - [Regularity of synaptic contacts forming in the molecular layer of the cerebral cortex after brief total brain anoxia]. AB - By the end of 10 and 90 min long total brain anoxia and on 1st, 3rd, 7th and 30th postanoxic days numerical density of synapses, shape and size of presynaptic dense projections (DP), curvature of synaptic contacts plane in molecular layer of cerebellar cortex were studied in control albino rats. Dependence between shape and extent of alterations of curvature of synaptic contacts and parameters and shape of DS was established. Maximum positive curvature of plane of synaptic contacts corresponded with maximum distance between DP, minimum height and spherical shape of DP. Maximum decline in numerical density of synapses and extent of curvature of synaptic contacts plane were found in zones of irreversible postanoxic damage of Purkinje cells. PMID- 12629801 TI - [Morphological and functional changes of nonapeptidergic cells in a surviving series of the rat hypothalamus under the effect of thyroliberin]. AB - Direct effect of thyroliberine on nonapeptidergic cells of supraoptic and paraventricular nucleus was studied in the experiment on surviving sections of hypothalamus. The inclusion of H3 thyroliberine into vasopressinergic cells and decrease of functional activity of vasopressin and oxytocinergic cells in both hypothalamic centres which is expressed in decrease of nucleolar size of these cells was demonstrated. PMID- 12629802 TI - [Local features of temporal organization of tissue mast cells in the dura mater of mature rat brain]. AB - Rhythms of functional activity of mast cells were studied in frontal, parietal and occipital areas of dura mater of mature rats. Chronological organization of mast cells dura mater was established. Circadian component dominated in all rhythms studied. Local peculiarities of mast cells rhythms were established; frontal area cells displayed most significant oscillations per day. They also showed maximum values of average day number and acrophase. Maximum functional activity was demonstrated in mast cells of frontal and parietal region one hour later than in parietal area of dura mater. PMID- 12629803 TI - [Information analysis of spinal ganglia]. AB - Information parameters (entropia and redundancy) of cervical and thoracic spinal ganglia of albino rat foetuses, mature animals (cat and dog) and human subjects were analysed. Information characteristics of spinal ganglia were shown to be level-specified and to depend on their functional peculiarities. Information parameters of thoracic spinal ganglia of man and different animals are specie specified and may be used in assessment of morphological structures as information systems. PMID- 12629804 TI - [Morphological and functional changes of tissue receptors of an organ plexus under the effect of blockers of cytoskeleton assembly]. AB - Colchicine and cytochalasine C exert modifying action on tissue receptors of urinary bladder of the frog. Under the effect of cytochalasine the period of vital staining of receptors were changed. Latent period of decoloration grew 37 56% longer while the period of staining accumulation grows 18% shorter. The forming of granules was also altered 2 types of terminal plaques were distinguished according to this sign in the course of treatment with colchicine and 3 types in the experiment with cytochalasine. Modifying action of cytochalasine was greater. Cytochalasin caused mass appearance of round intensely stained plaque the appearance of which was obviously connected with microtubules destruction. Mean area of terminals profile field also increased. Spontaneous impulse activity of receptors was significantly suppressed. Specificity of responses to colchicine and cytochalasine was probably conditioned by their effect on different elements of cytoskeleton. PMID- 12629805 TI - [Quantitative analysis of morphogenesis of local nerve plexuses forming in tissue culture]. AB - Discrete stages of formation of nervous plexuses in the culture of isolated neurons of a mollusc were analyzed autonomically. Area of plexus expansion (S1) and area, occupied by nerve structures (neurites and neuron bodies) were determined. It was demonstrated that despite intensive radial growth of peripheral neurites and S1 growth the density of plexus (S2/S1) was not changed. Relative acceleration of plexus expansion (as compared to S2 growth) registered on 40th and 50th hrs of cultivation indicates the absence of necessary cells in the plexus and development of contacts with adjacent local plexuses. Scanning of lamellar structures revealed submembranous aggregation of cytoskeleton apart from longitudinally directed cytoskeletal bands, which prevents one of the ways of neurite ramification. Elimination of lamellae between cytoskeletal bundles leads to formation of several neurites. PMID- 12629806 TI - [Development of the obturator artery in human prenatal ontogenesis]. AB - Definitive obturatory artery forms as a result of uneven growth of anastomosis of external and internal iliac arteries which is connected with peculiarities of regional organogenesis. 2 crucial periods--renal--and the period of gonade obstruction (the end of intrauterine development) were distinguished in morphogenesis of pubic anastomosis of iliac arteries. Sex related peculiarities of obturatory artery development were found in the second period. PMID- 12629807 TI - [Topographic features of the embryogenesis of human adrenal glands]. AB - The investigation of embryotopography of adrenals was performed on 95 embryo and 5-12 weeks old prefetus preparations, by microscopy, graphic and plastic reconstruction. It was abolished that adrenals develop in close morphologic correlation with temporary (primary kidneys, sex glands) and persistent (secondary kidneys) organs of embryonic retroperitoneal space. Developing in close correlation with derivatives of visceral layer of mesoderm (dorsal mesogastrium, mesocolon), adrenals, especially the left one are covered by an additional peritoneal layer during prefetal period. PMID- 12629808 TI - [Variants and regularities of hepatic artery topography (practical aspects)]. AB - Variants of forming of the arteries of upper abdomen were studied. Peculiarities of their location in different variants were considered using 75 visceral complexes. A description of the original variant of hepatic artery is given. 2 laws of arrangement of the arteries in the upper abdomen were formulated corresponding with all the anatomic variants known that explain predominant location of right and left hepatic arteries. PMID- 12629809 TI - [Significance of features of the projection of the cervical part of the thoracic duct onto the left sternocleidomastoid muscle for performing operations involving video support]. AB - To work out practical recommendations for using videosupport projection of cervical part of the thoracic duct on sternoclaidomastoid muscle was determined in 158 corpses of adults. Surgical access to the thoracic duct is available using common macrosurgical equipment until the moment of visualization of major lymphatic corrector, although it is reasonable to involve videosupport at following stage with the camera of videoendosurgical kit adjusted in inferior margin of the wound. This allows to perform all the necessary actions to expose cervical part of the thoracic duct and to correct them according to video information on the monitor. PMID- 12629810 TI - [Participation of a distant cholinergic mechanism in the response of the vascular bed to intoxication by organophosphorus inhibitors of cholinesterase]. AB - Poisoning with phosphacol causes dose--dependent decrease of blood flow speed in small vessels of mesoappendix. Administration of LD50 of phosphacole results in blood stagnation, simultaneous blood pressure fall, which leads to death of part of the animals. Electron microscopic study revealed the presence of acetyl and buthyryl cholinesterase in endotheliocytes of mesoappendicular capillaries, the activity of which was completely suppressed by administration of LD50 of phosphacol. 0,0-dimethyl-0 (2,2-dichlorvinyl) phosphate LD10 caused the damage of endotheliocyte surface. It was suggested that endothelial cholino-receptors that are activated through the rise of redundant acetyl-choline level in blood on the background of cholinesterase inhibition participate in the mechanism of pathological reactions described. Such variant of toxic effect was characterized as distant. PMID- 12629811 TI - [Cytochemical indices of blood lymphocytes in the assessment and prognosis of functional preparedness in a sportsman]. AB - Cytochemical criteria of assessment and prognosis of training were worked out. Correlations between lymphocyte dehydrogenase activity parameters and morphological content of blood, adipose and muscular mass and results were demonstrated. The study of lymphocyte dehydrogenase activity was significant for assessment of training efficiency. Enzyme profile of blood lymphocytes is an essential and prognostic sign of the state of compensatory adaptive reactions of the organism. Cytochemical methods of blood lymphocyte investigation provided stable and correct results that allowed to determine the onset of decompensatory process associated with nonrational training of any pathological condition and to reveal functional disorders at the level of the organism. Changes in lymphocyte enzymatic status in highly qualified sportsmen were a specific sign of adaptive reorganization of structures influenced by physical load. The use of parameters of blood lymphocyte dehydrogenase activity gives a possibility to fit an algorhythm providing stable prognosis of the state of the organism of sportsman throughout training during the year. PMID- 12629812 TI - [Morphological and functional characteristics of lymphocytes of mothers of children with Down syndrome]. AB - Nuclear chromatine of peripheral blood lymphocytes was studied in 13 women with children suffering from Down's syndrome using optic structural computer analysis. In 12 cases significant increase of nuclear roundness coefficient was determined. Deformation coefficient was determined for heterochromatine structures in 8 cases. Integral optic density of nuclear chromatine was significantly decreased in 12 women. This indicates the reduction of felgen-positive material due to deficiency of its compact fraction (in 11 cases). The activity of lymphocyte cytoplasmic lactate, alpha-glycerophosphate and succinate dehydrogenases (SDG) was studied morphocytochemically in 5 women who had children with the disease. High activity of mitochondrial SDG was determined in all cases which probably indicates disorders in lymphocyte energy state. This is one of the reasons for retention of risk pregnancy. Further research in this area may serve as a base for complete cytoanalysis in order to distinguish risk groups among women including primagravida for consequent determination of embryonal karyotype. PMID- 12629813 TI - [Morphological and biochemical changes in red and white muscle fibers in skeletal musculature ischemia]. AB - Structural reorganizations of red and white muscular fibres and peculiarities of changes in phosphoinositides in conditions of total ischemia of skeletal musculature were studied. On the whole morphological and biochemical changes were of the same type. However, decrease of phosphoinositides was more significant in white fibres than in red ones. In white muscular fibres deformations concerned myofibrillar apparatus while in red fibres mitochondrial apparatus was damaged. PMID- 12629814 TI - [Morphological characteristics of the glandular apparatus of the larynx in people of different ages]. AB - The larynx from 176 human corpses ranging in age from newborns to 90 year was examined macro-microscopically and histologically. In 1 period of maturity total number of glands varied from 116 man and from 110 to 587 in women. Density of glands with least length, width & thickness is maximal in the walls of the laryngeal ventriculus in comparison with vestibulum and cavitas infraglotis of the larynx. The density of the glands was smaller in newborns then in senile age people. PMID- 12629815 TI - [Age features of some structural characteristics of anal glands in children]. AB - Basing on macro-microscopical investigation of the anal canal of 24 children from newborns up to 12 years, structure and topography of the anal glands were studied. It was demonstrated that anal glands were arranged in rows or individually. Total number of anal glands varies from 8 to 12. The length of these glands major region increased from 0.41 mm in newborns to 0.84 mm in 8-12 years old children. In this period the total number of alveoli in gland's section increases from 7 to 18. Excretory ducts form flectures and open up in the bottom or in the walls of the anal sinuses. PMID- 12629816 TI - [Age and constitutional features in the forming of posture in children 7 to 14 years of age]. AB - Longitudinal study of forming of the posture was performed in 138 healthy pupils of Moscow school No. 710 aged from 6.5 to 14 of both sexes. Simultaneously children were examined anthropometrically and their constitution was determined after Schtefko. The posture was examined visually. It was established that posture forms within observation terms depending on sex, age and somatotype. Peak age for the risk of posture disturbance is 9-11 for girls and 10-12 for boys. Digestive and muscular somatotypes provide favourable prognosis respective to posture (in girls), while astenoid somatotype is the worst. Special type of posture defect is characteristic for each somatotype. In children with digestive and muscular somatotype primary posture defects disappear by 12-14. In children with astenoid and thoracal somatotype their number, in contrary, grows. PMID- 12629817 TI - [Method of topical diagnostics of morphological parameters of the thyroid gland]. PMID- 12629818 TI - [Morphological and functional characteristics of the lymphatic apparatus of the heart]. PMID- 12629819 TI - [Conference on "Current problems of operative surgery, surgical anatomy and pathophysiology"]. PMID- 12629821 TI - [110 years of the department of histology and embryology of the Siberian State Medical University]. PMID- 12629822 TI - CMS issues updated rules for the Medicare claims appeal process. AB - BIPA requires extensive changes to Medicare's appeal process. However, due to the lack of funding and adequate resources, CMS has been unable to immediately implement many of BIPA's far-reaching changes. CMS is committed to making a transition to the new process as quickly as possible and intends to implement as many key issues as feasible within budget and resource limits. Once Congress allocates funds for the new changes, CMS will be in a better position to implement all aspects of BIPA. [table: see text] The most significant change that BIPA presents is that Congress will require 12 new contractors to handle Part A and Part B appeals. These entities will be known as qualified independent contractors (QICs). CMS has completed development of the requests for proposals needed to solicit bids for the QIC contracts, including full statements of the scope of work (SOW) for these contracts. CMS released the draft SOW on Oct. 7, 2002. It is anticipated that the addition of the QICs will provide a more uniform and consistent handling of Medicare appeals on a national level. PMID- 12629823 TI - Medicare prescription plan, threat to AWP could impact the ESRD drug market. PMID- 12629824 TI - The ESRD regulatory landscape: a primer & resource. Part 2. PMID- 12629825 TI - "Giving back" on their own time for nearly four decades. PMID- 12629826 TI - New survey provides insight into CKD, exhaustion, anemia. PMID- 12629827 TI - Medicare medical nutrition therapy--one year later. PMID- 12629828 TI - [Dentin hypersensitivity and its significance in dental practice]. AB - Dentin hypersensitivity is not a new disease, more than a hundred years ago Gysi, later Brannstrom and many others were dealing with the problems of its symptoms, pathomechanism, differential diagnosis and therapy. The growing interest today maybe attributed to improving oral health, and to the presence of more teeth in older age, as a consequence of decreasing caries prevalence. The exposure of dentine surfaces due to gingival recession, erosion and abrasion may cause serious complaints, occurs in about 40% of the adult population, however, few of them turn to the dentist. Therefore, practising dentists should be aware of the possibilities of treatment, managing strategies and prevention. New aspects are the appropriate timing of toothbrushing after consumption of acidogenic, erosive foods and beverages, non-invasive (desensitizing, potassium-nitrate/fluoride containing toothpastes), and invasive (reconstruction with fillings, coverage of the exposed roots) treatment options. Continuous care of patients suffering in dentine hypersensitivity--in order to prevent more serious consequences (irritation of the pulp)--is advisable. PMID- 12629829 TI - [Correlation between the weight and usefulness of complete lower denture]. AB - The importance of gravity has been known for a long time in prosthetic dentistry. However, few researchers have focused on the measurement of the weight of complete lower dentures or on the possibility of increasing that weight and its effect on stability. Some authors recommend hidden weight increase using a horseshoe-shaped alloy covered by the artificial gum in order to increase the weight of complete lower dentures; others recommend the use of metal molars, heavy acrylic artificial gum, and a baseplate partially or fully made of metal for the same purpose. The authors of the present article have measured 167 complete lower dentures using an assay balance. They compared three sets of complete lower dentures of 24 patients, one of the three being of increased weight in each case. The examination by x2 statistics (four-domain contingency test) of the relationship between the weight and usefulness of complete lower dentures showed that an overwhelming majority of patients preferred the weight increased dentures. PMID- 12629830 TI - [Prosthodontic treatment in delayed pressure urticaria]. AB - The purpose of the presentation was to draw attention to a possible complication of a minimally invasive therapy outlining the complex role of the systemic etiologic factors. A patient with dermal pressure urticaria and polyvalent anaphylactic reactions showed up at our Dental Clinic. Urticaria was also present in the oral cavity. The main objective of the treatment plan was to find out how the edentulous alveolar ridge can be loaded and the selective support haw can be distributed on the abutment teeth. After the preventive and conservative phase of our comprehensive treatment lower and upper removable partial dentures were designed based on the selective support principles. In the maxilla a full denture while in the mandible a partial metal plate was made. After the metal framework having been tried in a second functional impression was taken to maximally utilize the advantages of the mixed (dental and mucogingival) support. The upper and lower alveolar mucosa could tolerate the denture plates after the insertion and only minute oedema occurred on the upper jaw along the post dam region which could be eliminated by small adjustments. It was concluded that in case of a known pressure urticaria the occlusal load of the edentulous mucosa can be kept on a tolerable level. Especially on the soft tissues with thick submucous layer can developed sever oedema (giant oedema, Quincke--oedema) that can lead to breathing difficulties, suffocation or even death. PMID- 12629831 TI - [Periodontal disease as a potential risk factor for preterm birth and low birth weight (Literature review)]. AB - In Hungary the number of low birth-weight new-born babies is about 8200 in a year, while the frequency of for preterm birth is 8.2%. The infant mortality among the low birth-weight babies is higher, than among the normal weight babies. Every efforts are used to decrease the number of preterm births. Researches have been carried out since decades on finding its possible risk factors. A possible risk factor for low birth-weight babies may be the chronic periodontal infection. The publication provides a review of recently published evidences of the potential association between periodontal infection and preterm low birth-weight. PMID- 12629832 TI - [Correction of maxillary atrophy with onlay-plasty]. AB - Following tooth loss, augmentation of the extensively atrophized alveolar ridge of the maxilla may be necessary, in order to restore the masticatory function by the replacement of teeth anchored on implants. A number of bone augmentation methods are available for the acceptance of implants. The aim of the presented report is to summarize our experience relating to the use of free monocortical bone grafts originating from the hip (spina iliaca anterior superior) for purposes of onlay-plasty. PMID- 12629834 TI - [Gardner syndrome. Case reports]. AB - The authors discuss 3 cases of Gardner-syndrome, outlining malignized large intestine polyposis, multiplex osteomas and the different skin tumours as the main features. The earliest symptoms suspecting the clinical picture are usually dental and/or mandibular lesions and draw the attention to the highly malignant polyposis. PMID- 12629833 TI - [Comparative evaluation of the shrinkage of addition-type silicone impression material using hand-mix and cartridge-mix technique]. AB - Impression materials show shrink during polymerisation. The average shrinkage of C-silicon impression materials is 0.6-1% after 24 hours. The A-silicons show 0.15 0.2% shrinkage after 24 hours. Like in other chemical reactions, the ratio of the ingredients and the mixing can strongly effect the final properties of the polymerised material. In their study the authors compared the shrinkage of Promodent ASK low-viscosity A-type silicon impression material using hand mix and cartridge mix technique. The measurements were carried out according to ADA 19 specifications. The statistical analysis of the results did not show significant differences in the shrinkage between the two mix techniques after 30 minutes, 24 hours and 72 hours. PMID- 12629835 TI - [Evaluation of manual dexterity in dental students at the Budapest University]. AB - Dentistry needs increased consideration in sensomotoric manipulation and coordinations of movements. The entrance procedure does not give any informations about ability of candidates. In history of Budapest University there were some shots about aptitude tests. The first tests were found out by Istvan Pesthy in 1963, which were testing only the manual skills, and were made by the passed-in students. The results did not affect the entrance. From 1974. Ferenc Molnar suggested a tentative manual test, which were compared with the other entrance tests results. The third generation of aptitude tests was not established, but the testing of manual skills and psychological aptness would be necessary in the entrance procedure. PMID- 12629836 TI - Nursing challenges and opportunities response to health needs in the community. PMID- 12629837 TI - REACH 2010: Charleston and Georgetown Diabetes Coalition approaches to reducing disparities for African Americans with diabetes. PMID- 12629838 TI - Community based telehealth: a future vision for South Carolina. PMID- 12629839 TI - Responding to bioterrorism and psychological toxicity: an introduction to the concept of shielding. PMID- 12629840 TI - Foundations of shielding. PMID- 12629841 TI - Public health and public trust: the defining dyad for the 21st century. PMID- 12629843 TI - Public participation and risk communication. PMID- 12629842 TI - Psychological challenges of bioterror: containing contagion. AB - In this article, we have introduced the concept of shielding as a public mental health intervention. Shielding addresses the core elements of bioterrorism when we consider that bioterrorism is best understood as terrorism, i.e., psychological warefare, which merely employs biologic agents, not to kill, but to terrorize. It is, therefore, to some degree dependent upon widespread contagion. Shielding is not a panacea. It is one aspect of an overall response plan. Nevertheless, it represents a potentially useful "antidote" for the bioterrorist assault. Perhaps most significant among its putative mechanisms of action appears to be controlling contagion, both physical and psychological. In the final analysis, in the wake of a terrorist attack, physicians can physically immunize and treat those who require such attention. Engineers can reconstruct buildings and roads. But who rebuilds the essence of humanity which has been violently ripped away from those who suffered the terrorist attack? How do we reconstruct a belief in justice and safety in the wake of a mass terrorist attack? Without attention to mental health, i.e., the "psychological side of terrorism," we run the risk of rebuilding a nation without a spirit, without a vitality, without a sense of humanity. PMID- 12629844 TI - Legal framework for shielding. PMID- 12629845 TI - Community shielding and the political system. PMID- 12629846 TI - The implementation of shielding. PMID- 12629847 TI - Smallpox outbreak: critical incident management. PMID- 12629848 TI - [Glaucoma and the quality of life]. PMID- 12629849 TI - [Retinal and retinal pigment epithelium hamartoma]. AB - Authors present four patients with combined hamartoma of the retina and retinal pigment epithelium who were observed for a period of 10 years. The significance of this entity lies primarily in its resemblance to malignant condition of the retina and choroid and in the vizual reduction. Clinical finding, diagnosis, natural history, histopathology, association with other diseases and therapy are discussed in this report as well. PMID- 12629850 TI - [Initial experience with implantation of miniaturized intraocular telescopes in dry type age-related macular degeneration]. AB - The authors evaluate the results of visual functions of atrophic form age-related macular degeneration of the retina by 4 patients after 18 months of the implantation miniaturized intraocular telescope. The improvement of the visual and subjective functions were evaluated in the most cases. PMID- 12629851 TI - [Effect of TLT absorptive eyeglasses on progression of myopia in children]. AB - The incidence of myopia is increasing worldwide. At the last VIIIth international conference on myopia held in June 2000 in Boston attention was aroused by the work of Quin et al. on the influence of noctural illumination in rooms where children slept on their subsequent development of myopia. Our working hypothesis presented in Hakone in 1996 presumes also a negative effect of light and heat which initiate the development of myopia but on eyes with an inborn disposition. We considered as a solution wearing spectacles with glass which allow to penetrate only visible light and absorb the other wavelengths. These glasses were supplied by Karel Kubena-TL Technologies. The authors followed up for four years two groups of children with progressive myopia. The progression of myopia during the first two years of the follow up was almost identical in the two groups: in the group with the absorption glasses 0.75 D/year and in the control group 0.77 D/year. After two years of wearing the absorption glasses the progression in the group with the absorption glasses was 0.34 D/year and in the control group 0.59 D/year. PMID- 12629852 TI - [Treatment of central serous chorioretinopathy with beta blockers]. AB - On a group of 13 eyes the authors evaluate their experience with medicamentous treatment of central serous chorioretinopathy. For treatment they used the non selective beta-blocker Trimepranol, 2 x 5 mg/day. They found that in 11 eyes, in 84.6% cases, adherence of the ablated neuroepithelium of the macula occurred latest within four months of treatment, in two eyes (15.4%) this therapeutic dose did not lead to remission of the disease even after four months of treatment and the condition was evaluated as failure of treatment. The authors conclude that a therapeutic dose of Trimepranol of 2 x 5 mg/day is not a reliable therapeutic solution of central serous chorioretinopathy. PMID- 12629853 TI - [Recurrent corneal erosion--current therapy]. AB - The work focuses on the present of the medical treatment of relapsing erosions of a cornea. It monitors the frequency of the occurrence of relapsing erosions of a cornea in our workplace in the ophthalmology department in Prague and possibilities of the usage of the therapeutic methods. The most frequent cause of the origin of relapsing erosion within our patients has been traumatic etiology of the cornea. From the results of our work complies that the most successful solution of this problem is the abrasion of the epithelium with the puncturation of the anterior stroma of a cornea and the phototherapeutic method "excime laser" (PTK). PMID- 12629854 TI - [Giant cell angiofibroma of the lower eyelids]. AB - The authors present a rare case of giant cell angiofibroma (GCAF) of the lower eyelids and the possibility of its surgical treatment. They emphasize the basic morphological indicators for assessment of GCAF and its difference from similar entities. PMID- 12629855 TI - [Assortative mating among individuals with refractive errors of the eye]. AB - Negative assortative mating for refractive errors of the eyes of 1241 married couples was discovered in the urban region of the South Moravian region. 46 married couples were myopic, 55 hyperopic, 73 astigmatic, 175 had combinations of refractive errors, 261 were emetropic and 631 were mixed. Negative assortative mating is highly significant (chi-square P < or = 0.001). It is more significant for urban population. Positive selection for emetropic married couples plays more important role in urban peoples than in citizens. PMID- 12629856 TI - [Use of Tanakan (EGb 761) in ophthalmology]. PMID- 12629857 TI - [Leukokoria--a serious symptom in pediatric ophthalmology]. PMID- 12629858 TI - [Detection of Chlamydial particles using direct immunofluorescence in ambulatory ophthalmologic practice]. PMID- 12629859 TI - [Jaroslav Kvapil, his eyes and his blindness]. PMID- 12629860 TI - Primary cutaneous B-cell lymphomas and pseudolymphomas: review of histopathological features. AB - The terminology and classification of lymphoproliferative disorders in the skin remains a source of confusion for clinicians and pathologists. More than 25 types of malignant lymphoid neoplasms may involve the skin. These either arise primarily or represent secondary cutaneous involvement due to the spread of node based lymphomas, other extracutaneous lymphomas or leukemias. In addition, there is a variety of cutaneous pseudolymphomatous reactions. This article addresses the histopathological features of primary cutaneous B-cell lymphomas and B-cell pseudolymphomas and presents a modern approach to the diagnosis of these lymphoid neoplasms. PMID- 12629861 TI - [Pathomorphology of the transjugular portosystemic shunt]. AB - An interventional radiological procedure TIPS (transjugular portosystemic shunt) assists in the decrease of portal hypertension. Portal blood is conducted via the hepatic vein into the lower vena cava. Blood drainage is performed via non vascular formation, i.e. by the channel created within the liver parenchyma (shunt). Histological changes (maturation) within the wall of an artificially formed blood conducting channel were studied on the base of examination of the set of 38 autopsy cases with the placed TIPS (25 males and 13 females, ages ranging from 22-81 years). Maturation course can be divided into three periods. The first stage is characterised by the presence of alternative changes mainly within the liver parenchyma surrounding the shunt. Reparative changes pass through the second stage with the predominance of liver cell debris removal and organisation of fibrin remnants. Consolidation changes dominate the third stage with the development of so-called neointima and giant-cell reaction around the stent struts. In three cases of the acute period of stent placement a new, so far not described specific phenomenon of compressive fusiform hepatocellular remodellation was found within the nodules of hepatocytes exposed to the pressure of resilient stent struts. Neointima is a non-thrombogenous structure participating on the shunt patency. It is possible to judge approximately the time of stent placement according to the histologically estimated level of shunt wall maturation. TIPS is a long-lasting supplement conduit between the portal and systemic blood circulation. Contrary to the easily reconstructable formal pathogenesis of histological changes, the problems of causal pathogenesis of the neointima challenge mainly the interest of laboratories involved in basic science. PMID- 12629862 TI - [What is your diagnosis? Answer: nuchal-type fibroma]. PMID- 12629863 TI - [Metaplastic spindle-cell (fibromatosis-like) carcinoma of the breast--report of 4 cases]. AB - The authors report about clinico-morphological features in four cases of spindle cell metaplastic carcinoma of the breast, closely mimicking benign soft tissue fibromatosis or nodular fasciitis. All patients were females aged 54-72 years with a palpable nodule within the breast, 20-35 mm in size. Histologically, the tumor infiltrated surrounding tissue; it consisted of spindle cells with relatively bland morphology, arranged in a fascicular, storiform or random fashion. Mitotic activity was low, focally the tumors revealed regressive changes (hyalinization, myxoid degeneration). Cytokeratin expression was proven in all four cases. In two patients the tumor metastasized and the patients died of it; this fact confirms recently published data that despite its deceptively benign appearance, fibromatosis--like carcinoma of the breast is undoubtedly a malignant process with potentially aggressive behaviour. Differential diagnosis of this unusual variant of metaplastic breast carcinoma is discussed together with related therapeutic issues. PMID- 12629864 TI - Intestinal necrosis following Calcium Resonium-sorbitol administration in a premature uraemic infant. AB - Sodium polystyrene sulphonate (Resonium A) in sorbitol given as an enema or orally to treat hyperkalaemia has been described to induce intestinal necrosis in uraemic patients. We report a case of a premature infant with acute renal insufficiency who developed focal transmural necrosis and perforation of the small intestine after 10 days of administration of calcium polystyrene sulphonatum (Calcium Resonium) in sorbitol by enema and by nasogastric tube. On histological examination of the resected part of the small intestine, numerous strongly basophilic angular crystals of resonium were found in the lumen, in the necrotic wall, as well as in the organized exudate on the peritoneal surface. The crystals showed a strong direct Schiff positivity without preoxidation. They were also stained using PAS, Giemsa, Ziehl-Neelsen, Schmorl, and Gram method. In contrast, the crystals were Congo red and Alcian blue (pH 2.5) negative and non birefringent. The direct Schiff positivity without preoxidation is virtually pathognomonic for resin crystals in routinely processed tissues. The same crystals were observed in the lumen of the small intestine and in peritoneal adhesions at autopsy. Thus our case provides additional evidence that Resonium A/Calcium Resonium in sorbitol administered as an enema or orally can lead to intestinal necrosis in uraemic patients. PMID- 12629865 TI - Composite dermatofibrosarcoma protuberans-giant cell fibroblastoma recurring as Bednar tumor-giant cell fibroblastoma with mucoid lakes and with amputation neuroma. AB - We report an unusual case of composite giant cell fibroblastoma dermatofibrosarcoma protuberans (DFSP) that, in its second recurrence, contained a pattern of Bednar tumor (BT) and giant cell fibroblastoma (GCF). The recurrent tumor showed extreme myxoid change with creation of mucoid lakes, which mimicked a pattern of myxoid liposarcoma. One area in the recurrent lesion contained amputation neuroma overgrown with neoplastic spindle cells, which simulated a nerve sheath neoplasm. This case demonstrates common histogenesis of GCF, DFSP and BT, and it shows how broad morphological spectrum can be produced by a composite tumor, especially when the tumor includes unconventional growth pattern or additional non-neoplastic lesion. PMID- 12629866 TI - [Solid and papillary epithelial tumor of the pancreas: cytologic and histologic features]. AB - Cytologic and histologic features in a case of solid and papillary epithelial neoplasm (SPEN) of the pancreas in a 34-year-old female are presented. In the fine-needle aspiration material, there was a predominance of pseudopapillary fragments with delicate fibrovascular core lined with one or more layers of uniform tumor cells with clear cytoplasm. There were also acinar-like or rosette like structures and dispersed tumor cells. Tumor cells had small, round to oval nuclei, with finely granular chromatin and inconspicuous nucleolus. Rarely, nuclear grooves were present. Mitoses were not seen. Histologically, the encapsulated tumor was predominantly solid, with focal pseudocystic and pseudopapillary areas; foreign body granulomas were focally present. Tumor cells were small and uniform with clear cytoplasm. Mitoses were extremely rare. Immunohistochemically, the tumor cells revealed diffuse positivity of vimentin, and focal positivity for alpha 1-antitrypsin and CD68. No immunoreactivity for chromogranin, synaptophysin, cytokeratin, HMB 45, estrogen and progesterone receptors was found. In differential diagnosis, it is important to distinguish SPEN mainly from mucinous tumors of the pancreas, microcystic adenoma and pseudocysts, but also from endocrine tumors of the pancreas, acinic cell carcinoma, renal cell carcinoma, adrenal cortical adenoma and primary extrapulmonary sugar tumor. PMID- 12629867 TI - [Pseudomalignant changes in benign phyllodes breast tumor]. AB - Phylloides tumor of the breast is well known entity in pathology for a long time. It includes fibroepithelial tumors of the different biological character. They are generally divided into three groups: benign, low-grade malignant (borderline) and high-grade malignant. Adenoses of the breast are a group of the benign lesions, the main feature of which is the proliferation of the glandular component of the breast. More frequent adenoses occurring in practice are: blunt duct adenosis, sclerosing adenosis, nodular adenosis, adenosis tumor and florid adenosis. The less frequent adenoses are: microglandular adenosis, tubular adenosis, apocrine adenosis, and myoepithelial (adenomyoepithelial) adenosis. In this article two histologically unusual lesions of the breast in middle-aged women are presented, consisting of the benign variants of the phylloides tumor, modified distinctly by various types of adenoses, which can cause, together with intensive myoepithelial proliferation, great diagnostic problems, especially on frozen sections. They may be mistaken with invasive ductal or lobular cancer. Reliable differentiation between these entities is possible only by using of the immunohistochemical methods. PMID- 12629868 TI - [Findings in myocardial extracellular matrix in fatal poisoning with addictive substances]. AB - It have been reported that chronic drug abuse induces changes in extracellular matrix (ECM) composition of myocardium. Matrix metaloproteinases (MMP) play important role in ECM remodelling and alteration in the ECM of myocardium. They are able to cleave ECM components and may contribute to progression of heart damage. Remodelling implies an alteration in the ECM and in the spatial orientation of cells and intercellular components. In this report we evaluated the extracellular matrix of myocardium of deceased drug abusers and controls. The myocardium of 5 toxicologically positive cases and 4 control cases were analysed by zymography of MMP, electrophoresis of collagens and histologicaly. Gelatinases (MMP-2 and MMP-9) and collagenase (MMP-1) activities were increased in drug abusers compare to the controls. In contrast, alpha chain of collagen type I after 160 hours collagen extraction, were not observed in control specimens. The activation of MMP proenzymes is one of the critical steps that lead to ECM breakdown. Our findings suggest that the myocardial collagen matrix is damaged in drug abusers. PMID- 12629869 TI - [Frequency of occurrence of traumatic aortic rupture in automobile passengers involved in a frontal crash with a static or dynamic obstacle]. AB - There is a lesion of aortic thoracic complex in car-passengers during the frontal clash to a static or to a dynamic antipodal obstacle is described in this study. It is a retrospective analysis. Facts were noted especially from dissectional documents. All from 298 men died. The main cause of car-passengers death was a weighty traumatic lesion of cardiovascular system in 58.7%. The aortic rupture was noted in 98 (32.9%) people, the death was in 90.8% directly on the road and 9.2% at hospital. From the whole people who were accepted to a hospital were 9 (8.9%) with a traumatic aortic rupture. But nobody of them was transported to a special center and everyone died due to traumatic lesion of cardiovascular system. Statistical significant cofactors of aortic rupture were atherosclerosis of aortic wall and intensity of clash. Therefore we can expect an aortic rupture in every third dead frontal car crash participant on a dynamic or static obstacle. Nearly 10% from men with traumatic aortic rupture were transported to a hospital. No aortic rupture was diagnosed. PMID- 12629870 TI - [Terminology used in evaluating the severity of injuries for agencies involved with the penal system]. AB - The authors deal with medical and juristis terms concerning the evaluation of the grievousness of injuries for the police of the Czech Republic. PMID- 12629871 TI - [How a young physician developed important forensic medical evidence. On the 130th anniversary of the birth of Dr. Ivan Halek (1872-1945)]. PMID- 12629872 TI - A new approach to testing the effects of group psychological therapy on length of life in patients with metastatic cancers. PMID- 12629873 TI - Psychosomatic integrative treatment and rehabilitation. PMID- 12629874 TI - Limitations of randomized clinical trials in chronic diseases: explanations and recommendations. PMID- 12629875 TI - Psychosocial genomics: gene expression, neurogenesis, and human experience in mind-body medicine. AB - This paper introduces the new scientific discipline of psychosocial genomics as an emerging area in the foundations of mind-body medicine. Psychosocial genomics brings together a variety of interdisciplinary fields ranging from studies of stress, psychosomatics, psychoimmunology, and psychoendocrinology to the deep psychobiology of creativity, optimal performance, dreaming, art, ritual, culture, and spiritual life. The focus of psychosocial genomics is on creating new models of how the levels of gene expression, neurogenesis, and healing are interrelated as a complex, adaptive system with the levels of human experiencing, behavior, and consciousness. Ongoing research in psychosocial genomics is presented to illustrate how this complex adaptive system operates in the theory and practice of alternative and complementary mind-body medicine and psychotherapy on all levels, from mind to gene. PMID- 12629876 TI - Revisiting the health history: an integrative medicine approach. PMID- 12629877 TI - Investigation of an E. coli O157:H7 outbreak in Brooks, Alberta, June-July 2002: the role of occult cases in the spread of infection within a daycare setting. PMID- 12629878 TI - Burden of proof. PMID- 12629879 TI - Minority report. PMID- 12629880 TI - Home and away. PMID- 12629881 TI - Keeping it real. Interview by Steven Black. PMID- 12629882 TI - Double trouble. PMID- 12629883 TI - All together on hygiene. PMID- 12629884 TI - Good grief. PMID- 12629885 TI - Poor relations. PMID- 12629886 TI - Bringing health care to schools. AB - AIM: To explore secondary school students' health knowledge, attitudes and behaviours, and to examine staff assessment of the Bodyzone project, in which confidential drop-in clinics for young people are set up at schools. METHODS: A cross-sectional study involving a questionnaire, participatory group assessments and staff interviews was used to gather data from students and staff at two schools for phase one of the evaluation. RESULTS: The results showed that the Bodyzone service is valued by those who use it and by school staff. CONCLUSION: The data suggest that the Bodyzone programme has many of the elements of a successful adolescent healthcare service and deals with a range of health and social problems experienced by young people. However, its small scale of operation hampers the capacity to provide an effective service to all who might need to access it. PMID- 12629887 TI - Implementing a critical care course for ward nurses. AB - This article describes the setting up and evaluation of a course for ward-based nurses, which is designed to improve their skills in caring for highly dependent patients. The authors suggest such courses can increase nurses' confidence and competence and, as well as enhancing care, can improve communication with senior staff about issues of concern. PMID- 12629888 TI - Assignment writing skills. AB - This article offers students on diploma and degree courses a constructive step-by step guide to planning, writing and critically reviewing their module assignment papers. Guidance is accompanied by a selection of practical activities designed to enhance students' information gathering and assignment writing skills. PMID- 12629890 TI - Postnatal testicular development, cellular organization and paracrine regulation. PMID- 12629889 TI - Male sex determination and prenatal differentiation of the testis. PMID- 12629891 TI - Regulation of spermatogenesis. PMID- 12629892 TI - Male germ cell apoptosis. PMID- 12629893 TI - Gonadotrophin actions on the testis--genotypes and phenotypes of gonadotrophin and gonadotrophin receptor mutations. PMID- 12629894 TI - Physiology and disorders of testicular descent. PMID- 12629895 TI - Testicular neoplasia in childhood and adolescence. PMID- 12629896 TI - Testicular function after cancer treatment in childhood. PMID- 12629897 TI - Present and future options for the preservation of testis tissue and function. PMID- 12629898 TI - [The new laws defining the supply of care in terminal renal failure]. PMID- 12629899 TI - [Iron, oxidative stress and infection in hemodialysis]. PMID- 12629900 TI - [Nutrition and renal failure: what's new?]. PMID- 12629901 TI - [Evaluation of the needs of a population of hemodialysis patients in Rhone Alpes]. AB - The evaluation of the needs of patients with terminal renal insufficiency is necessary to determine the appropriate care programme. An evaluation of the medical and nursing needs, as well as those related to handicap (a total of 47 items) was undertaken on all hemodialysis patients in the Rhone-Alpes region. Only 34.4% were free of co-morbidity and 35.7% of significant handicap, 63.7% needed no other nursing care than that related to supervision of the dialysis and 16.9% were free of all the conditions studied. The patients treated in the centre required care needs than those treated in all other structures. The autodialysis population differs from that treated at home by a greater need for care due to handicaps, and from that treated in a dialysis centre by a lesser degree of handicap and less need for medical treatment. A level of theoretical unsuitability (needs incompatible with the statutory provisions for each type of treatment) was defined: it concerns 5% of the population in a dialysis centre and to 33% of the population on home dialysis. The needs evaluation grid could be used to quantify the requirements of a dialysis population and to arrange for the necessary resources. PMID- 12629902 TI - [A tool to predict the resources necessary for the whole hemodialysis population]. AB - Predicting the resources necessary for the treatment of terminal renal insufficiency requires an understanding of the needs of the whole dialysis population. This study evaluates the advantages of a complete evaluation grid for care needs (CG, 47 items) compared with a simplified version (SG) and with the data obtained from REIN (R) as a tool for predicting the required resources. Compared to CG, the two other classifications under-estimate the level of care needed and the prevalence of patients who have at least one condition resulting in an 'excess' of needs. In a system with three types of structures, the theoretical distribution of patients according to the CG is: in structure A (permanent medical presence): 43.4% (vs 39 for SG and 21% for R); in structure B (intermediate): 34% (vs 31.5 and 24.7% respectively); and in structure C (independent): 22.3% (vs 29.5 and 53.5%) of patients. The care needs scores of populations assigned by R to home dialysis are incompatible with the resources allocated to it. With a certain number of adjustments and qualifications, a population study using a simplified needs grid could enable a prediction of the resources which need to be provided. The patients' records which take precedence in the future REIN should be modified in order to make possible a regular evaluation of the appropriateness of the resources allocated. PMID- 12629903 TI - [Ossifying fibroma of the face and hyperparathyroidism in a chronic hemodialysis patient]. AB - The bony complications in secondary hyperparathyroidism in chronic renal failure are varied. The ossifying fibroma is a rare manifestation. We report a case of a 40 years old woman with indeterminate nephropathy undergoing hemodialysis since 1982. Since may 1995, the patient had a diffuse and intensive bone pain of the two inferior members inducing functional importance with apparition of endobuccal tumefaction in the hard palate with an important increase of the volume of the maxillo-mandibular complex six months later associated to disturbances of the deglutition and phonation. In december 1995, the serum parathyroid hormone level is measured at 1527.6 pg/ml and the serum alkaline phosphatase level at 1620 UI/l. The bony lesions are disseminated to all the skeleton in the form of a demineralization with an ossifying expansive process affecting the maxillo mandibular complex. Calcemia was at 2.3 mmol/l and phosphoremia at 2.1 mmol/l. The surgical biopsy of this pseudotumor showed an aspect of ossifying fibroma. The cervical echography showed a left inferior parathyroid nodular. In june 1996, a subtotal parathyroidectomy was done. The 9th day after surgery, the serum parathyroid hormone level was at 103 pg/ml with normal calcemia and phosphoremia. Five years later, the volume of the tumor reduced moderately. This observation induces 3 commentaries: the bony manifestations associated to secondary hyperparathyroidism have sometimes a tumoral aspect, the ossifying fibroma may be for known as well as the brown tumor and the treatment of these tumors in complex justifying furthermore a rigorous prevention of the hyperparathyroidism in hemodialyzed patients. PMID- 12629904 TI - [Falling in the elderly, does a 'falling clinic' make sense?]. PMID- 12629905 TI - [Serum markers in relation to cognitive functioning in an aging population: results of the Maastricht Aging Study (MAAS)]. AB - Little is known of the biochemical processes of cognitive decline during 'healthy' aging. Biological markers in body fluids, such as blood, could provide insight in those processes. In the present studies serum concentrations of different markers have been correlated to cognitive functioning of cognitively healthy aging individuals over a period of six years (mean age 57 years, SD 11, n = 93). Markers were related to mechanisms known to be involved in Alzheimer's disease, including inflammation, cholesterol homeostasis and homocysteine homeostasis. Domains of cognitive function addressed were cognitive speed (Letter Digit Coding test), attention and information processing (Stroop test), and memory (Word Learning test: Total Words and Delayed Recall). Baseline concentrations of haptoglobine, homocysteine, lathosterol and lanosterol were negatively correlated with cognitive functioning on the Stroop test over the whole follow-up period of six years. Concentrations of all markers, i.e. haptoglobine, C-reactive protein, homocysteine, lathosterol and lanosterol, were also negatively correlated with functioning on the Word Learning test (Delayed Recall and for some markers also with the Total Words) over the whole six-years follow-up period. In conclusion, concentrations of serum markers related to inflammation, homocysteine and cholesterol homeostasis are not only associated with Alzheimer's disease, but also with cognitive functioning in the cognitively healthy aging population. PMID- 12629906 TI - [Alzheimer's disease. Present and future role of genetics]. AB - Research in the field of Alzheimer's disease has shown that genetic factors play an important role in the aetiology of the disease. Until now, four genes have been found to be implicated in Alzheimer's disease. Mutations in the amyloid precursor protein gene (APP) and the presenilin genes (PSEN1 en PSEN2) cause early onset Alzheimer's disease. These mutations segregate in an autosomal dominant pattern. The fourth gene involved in Alzheimer's disease is the apolipoprotein E gene (APOE). Carriers of the E4 variant of APOE have an increased risk of Alzheimer's disease. Being a carrier of this E4 variant increases the risk of both early- and late-onset Alzheimer's disease. Of the four Alzheimer-genes, APOE plays the most important role in the general population. Mutations in APP and the presenilin genes account for less than 1% of the prevalence of the disease in the general population compared to 10-17% for the APOE variation. Up till now the impact of genetics in daily clinical practice is very limited. However, genetics has caused major progress in molecular-biological knowledge, especially of the amyloid metabolism, creating optimism about novel biological markers and eventually therapeutic strategies. In Alzheimer's genetics break-throughs are to be expected using classical methods such as the candidate gene or linkage approach. Novel strategies such as genetic research in isolated populations are promising. PMID- 12629907 TI - [Care quality: reliability and usefulness of observation data in bench marking nursing homes and homes for the aged in the Netherlands]. AB - Before including quality of care indicators in the Benchmark of Nursing Homes and Homes for the Aged in the Netherlands the reliability of the patient data collection, and usefulness had to be established. The patient data items were derived from the Resident Assessment Instruments (RAI) and a questionnaire on social interaction in elderly people. Three nursing homes and three homes for the aged participated in the test with 550 patients. 279 x 2 assessments were collected by independent raters for an inter rater reliability test; 259 x 2 by the same rater for a reliability test-retest; and 24 by a single rater. The scores on paired assessment forms were compared with the weighted Kappa agreement test. The test results allowed 10 of the 13 quality indicators from RAI to be retained. In addition new quality indicators could be defined on 'giving attention' and 'unrespectful addressing'. We estimate on the basis of a questionnaire for the raters that on average 9 to 12 minutes per patient are needed to collect and enter data for the resulting 12 quality indicators. PMID- 12629908 TI - [Prevalence of xerostomia and hyposalivation in the nursing home and the relation with number of prescribed medication]. AB - In nursing homes, the number of patients with their own dentition increases. Many of them, however, are unable to perform oral hygiene procedures. Medication is considered to be an important cause of hyposalivation, which may lead to oral alterations and loss of teeth. The aim of the present study was to investigate the prevalence of xerostomia and hyposalivation in a Dutch nursing home and to examine the possible relation with medication usage. Between January and March 2001, the salivary flow rates were measured in 50 patients residing in a nursing home in Amsterdam. Unstimulated saliva, parafilm-stimulated saliva and citric acid stimulated saliva were determined. Xerostomia was determined by the question "My mouth feels dry" and the medication used was examined. The data were analysed with (M)ANOVA. The average age of the patients was 78.1 +/- 9.7 years. Forty eight % of the patients had an unstimulated flow rate of less then 0.20 ml/min and 24% had a flow rate even lower than 0.10 ml/min (reference values: 0.25-0.50 ml/min). The flow rate of women was significantly lower than that of men (p < 0.01), even after correction for age, smoking and the number of prescribed medications. Salivary flow rates decreased significantly with age (p < 0.05). The number of prescribed medication was significantly higher in patients over the age of 70 (p < 0.01, n = 42) and also in women (p < 0.01). The prevalence of xerostomia was 52% with no difference between men and women. In nursing homes, the prevalence of hyposalivation and xerostomia is high. The number of xerogenic medications used seems to be an important factor. Women and patients aged over the age of 70 years need special attention with respect to oral health. PMID- 12629909 TI - Training the public health practitioner. PMID- 12629910 TI - Need for physicians trained in preventive medicine and public health: implications for a bioterrorism response. PMID- 12629911 TI - Strengthening the public health workforce: three CDC programs that prepare managers and leaders for the challenges of the 21st century. AB - To address the need for management development in public health, the Centers for Disease Control and Prevention (CDC) established three independent workforce development initiatives aimed primarily at strengthening management and leadership capacity: the Sustainable Management Development Program, the Management Academy for Public Health, and the CDC Leadership and Management Institute. Though independently designed and implemented, the programs share similar guiding principles in their approach to management development: interactive (adult) learning, management tools that reinforce evidence-based decision making, individual feedback, continuous improvement of the learning process, posttraining support for networking and life-long learning, and teamwork. This article will discuss important lessons learned regarding best practices in management and leadership development. PMID- 12629912 TI - Does leadership training make a difference? The CDC/UC Public Health Leadership Institute: 1991-1999. AB - Public health leadership development programs have proliferated since the release of the Institute of Medicine's call for strengthened public health leadership. Little has been documented, however, about the impact of these programs. This article presents results of an eight-year retrospective evaluation of the Centers for Disease Control and Prevention/University of California Public Health Leadership Institute, the nation's first year-long leadership development program serving senior public health leaders. Results show that this program has had a positive impact on participants' leadership effectiveness at the personal, organizational, and community levels as well as on the field of public health. PMID- 12629913 TI - An interdisciplinary MPH via distance learning: meeting the educational needs of practitioners. AB - This article describes a distance learning master of public health program that prepares students to lead programs and organizations. Evaluation showed that the curriculum, format, and teaching methods were accessible, affordable, acceptable, and appropriate for the working professional, and equivalent to residential master's programs aimed at experienced professionals. Students interacted with professors and other learners using the World Wide Web, weekly videoconferences, and face-to-face meetings. Students, whose main motivations for enrolling included improving job performance in leadership and career advancement, reported strengthening of knowledge, perspective, skill, technical facility, confidence, and job performance. Learner satisfaction was high, with 100% agreeing they would recommend the program to others. PMID- 12629914 TI - Educational partnerships for public health: do stakeholders perceive similar outcomes? AB - Based on a study of five W.K. Kellogg Foundation-funded educational partnerships, this article reports the stakeholders' views on five sets of anticipated outcomes: health professions education impact, curricula and services, students, community and policy, and sustainability and structural change outcomes. The participants had a reasonable degree of certainty that their partnerships would achieve the intended outcomes. Using tightly defined "specific" stakeholder groups for the analyses reflected a more precise picture than employing "generic" groups. Partner groups that perceived greater certainty about the outcomes had higher scores on other related partnership parameters. For partnership working, the article proposes an orbital hypothesis of stakeholders' perceptions for further investigation. PMID- 12629915 TI - Identifying training needs in the public health workforce: the public health prevention service as a case study. AB - The Public Health Prevention Service is a three-year training program committed to developing a workforce that is skilled in planning, managing, and evaluating prevention programs. In 2000, a questionnaire was administered to participants to assess their training needs. According to the resultant data, training is preferred in traditional formats (e.g., case studies and group discussion) and in short installments (one to three hours). Topics identified for future training included software application, prevention effectiveness (i.e., economic analysis), program evaluation, budgeting, and consensus building. PMID- 12629916 TI - Supporting academic public health practice: a survey of organizational structures in public health schools. AB - A survey was conducted at 28 schools of public health to assess how they allocate responsibility for three kinds of academic practice activity: continuing education, student internships, and technical consultation by faculty. Respondents identified whether these responsibilities were in a central practice unit, in another schoolwide unit, dispersed among several units, or dispersed among individual faculty members. Though all respondents engaged in these activities, responsibility tended to be carried out by individual departments or independent faculty members. This typically decentralized approach raises questions about the sustainability of academic public health practice that call for further inquiry. PMID- 12629917 TI - Practice rewards in academia. PMID- 12629918 TI - The tenure dilemma: furthering the discourse. PMID- 12629919 TI - Variability in diagnostic criteria for chronic fatigue syndrome may result in substantial differences in patterns of symptoms and disability. AB - Chronic fatigue syndrome (CFS) is an illness that involves severe, prolonged exhaustion as well as neurologic, immunologic, and endocrine system pathology. Because the pathogenesis of CFS has yet to be determined, case definitions have relied on clinical observation in classifying signs and symptoms for diagnosis. The current investigation examined differences between CFS as defined by Fukuda and colleagues and a set of criteria that has been stipulated for myalgic encephalomyelitis (ME). Dependent measures included psychiatric comorbidity, symptom frequency, symptom severity, and functional impairment. The ME and Fukuda et al. (1994) CFS criteria were compared with a group having chronic fatigue due to psychiatric reasons. Significant differences occurred primarily with neurologic, neuropsychiatric, fatigue/weakness, and rheumatological symptoms. These findings suggest that it might be inappropriate to synthesize results from studies of this illness that use different definitions to select study populations. PMID- 12629920 TI - Impact of differential response rates on the quality of data collected in the CTS physician survey. AB - Survey administrators face trade-offs between expending additional survey resources to maximize response rates versus using fewer resources and accepting lower response rates. Using data from the Community Tracking Study's Physician Survey, we examined how survey estimates and data quality changed as additional respondents completed the survey. Results showed that improvements in response rates over the range examined (i.e., up to 65%) did not change estimates appreciably nor affect data quality. As long as these results are not overstated to imply that extremely low response rates are credible, this study may permit researchers to disseminate interesting results in peer-reviewed journals even when the response rate falls slightly short of current standards. It must also be emphasized, however, that we were unable to measure the nonresponse effect of those who were never interviewed. Achieving a response rate significantly above 65% might have changed the survey results appreciably. PMID- 12629921 TI - Assessment of tuberculosis treatment completion in an ethnically diverse population using two data sources. Implications for treatment interventions. AB - Many adolescents who are prescribed therapy for tuberculosis (TB) infection fail to complete it. This article presents the results of a demographic and epidemiological assessment of TB treatment completion in adolescent populations using (a) surveillance data from the Los Angeles Health County Department and (b) a prospective/retrospective medical chart review from targeted clinics. Patients who did not complete the 6-month recommended medical treatment for latent tuberculosis therapy (LTBI) averaged 13 weeks in care. Younger age (OR = 1.15; 95% CI 1.03-1.29), birth in the United States (OR = 1.92; 95% CI 1.14-3.22, compared to Mexico), and Asian ethnicity were associated with completion of care. In multiple logistic regression analysis, age (OR = .88; 95% CI .78-.98) and Latino ethnicity (OR = .53; 95% CI .29-.95) remained significant predictors of completion of treatment. These findings indicate the need for age-specific educational reinforcement and cultural differentials in completing care for LTBI. PMID- 12629922 TI - A validity study of expert judgment procedures for setting cutoff scores on high stakes credentialing examinations using cluster analysis. AB - This study compares an expert judgment process--minimal performance levels (MPL) using the Nedelsky and Ebel procedures--for setting cutoff scores for pass/fail on licensure examinations with an empirical approach--cluster analysis. Data from all three components of the Canadian Standard Assessment in Optometry (CSAO) examinations (knowledge, clinical judgment, and clinical skills) from 243 candidates were obtained. Results indicate that for the written components of the exams employing the Nedelsky method of MPL setting, there was a mean agreement of pass/fail of 81% with the cluster analysis approach on pass/fail categorization. For the performance exams using the Ebel method, the mean agreement of pass/fail with the cluster analysis was 93%. Thus the subjective approaches to setting cutoff scores (i.e., expert judgment methods) converge with the objective method (i.e., cluster analysis) of classifying test takers in the same categories. PMID- 12629923 TI - Paraphrase procedures for assessing comprehension of health outcome measures: an illustration from schizophrenia research. AB - Valid measurement of health outcomes require instruments that are meaningful to those who use them. Clinicians, patients, and others involved in health care have different perspectives on health conditions that are likely to lead to different interpretations of outcome measures and reductions in the accuracy and validity of outcome measurements. This study used paraphrase procedures, a systematic approach for evaluating respondent comprehension, to evaluate descriptions designed for use in studies of schizophrenia treatment outcomes. In four stakeholder groups directly and indirectly involved in schizophrenia treatment (clinicians, patients, patients' families, and the general public), the paraphrase procedures identified comprehension problems that could compromise measurement quality and yielded specific information to guide improvement of the outcome descriptions. The paraphrase procedures detected comprehension problems that were not identified by respondents. This study suggests that systematic assessment of comprehension can improve the quality of health outcome measurement. PMID- 12629924 TI - Validation of an observation instrument for measuring student engagement in health professions settings. AB - Documenting student engagement has received increased emphasis in medical schools, as teaching strategies are changing to include more student-to-student interactions. The purpose of this study was to develop and evaluate a measure of student engagement completed by independent observers that would not interfere with student learning time. Data from 3,182 observations completed by nine observers in 32 educational classroom settings with 23 different instructors were used to evaluate the interobserver reliability and gather validity evidence for our observational instrument, named the STROBE. Results indicated that interobserver agreement was good to excellent when observations were conducted simultaneously on randomly selected students in the same classroom (84% average agreement and 0.79 average kappa coefficient) and when observations were conducted on different randomly selected students (79% average agreement). Results also provided strong evidence for validity. Overall, findings indicate that the STROBE demonstrates promise for educational research and evaluation by documenting student engagement in medical education settings. PMID- 12629925 TI - Development of the approaches to work and workplace climate questionnaires for physicians. AB - Two questionnaires were developed to investigate the workplace learning of physicians. The Approaches to Work Questionnaire for Physicians and the Workplace Climate Questionnaire for Physicians were adapted from general measures developed by Kirby, Knapper, Evans, Carty, and Gadula. These questionnaires were administered to a random sample of Ontario physicians. Consistent with the results of Kirby et al., three dimensions of approaches to work were observed: Deep. Surface-Rational, and Surface-Disorganized. Three dimensions of workplace climate were also found, Supportive-Receptive, Choice-Independence, and Workload. Results indicate that physicians adopt primarily a Deep approach to work, but that there is a smaller tendency toward Surface-Disorganized learning, one that is strongly correlated with perceptions of heavy workload. The Deep approach was associated with work environments perceived to be Supportive-Receptive and offer Choice-Independence. The use of these questionnaires in research and practice is discussed. PMID- 12629927 TI - The social impact of medicine. Chapter 3. PMID- 12629928 TI - The joy of a medical career. PMID- 12629926 TI - Clinical Linguistics and Phonetics' first 15 years: an introductory comment. PMID- 12629929 TI - A new year, a new look. PMID- 12629930 TI - Reader updates guidelines for I-131 and Y-90. PMID- 12629931 TI - Cyclooxygenase-2: from arthritis treatment to new indications for the prevention and treatment of cancer. AB - The discovery of the isoenzymes cyclooxygenase-(COX-) 1 and COX-2 led to the development of newer nonsteroidal anti-inflammatory drugs (NSAIDs) designed to block COX-2, such as rofecoxib, celecoxib, and valdecoxib. Because of the specificity of COX-2 expression, COX-2 inhibitors have the potential to reduce the risk of gastrointestinal bleeding experienced with the use of classic NSAIDs. With their crucial role in the control of inflammation, the COX-2 agents originally were marketed for the treatment of rheumatoid and osteoarthritis. However, promising new indications for COX-2 agents in the prevention and treatment of cancer are under investigation. The role of aberrant COX-2 expression in the development of cancer has been studied most widely in patients with colon cancer and adenomas. Recent studies suggest that COX-2-derived prostaglandins may play an important role in tumor viability, growth, and control of metastasis. Possible new indications for the use of COX-2 inhibitors to prevent and treat cancers may be monumental. However, therapy with these agents is not without risk. Oncology nurses must be aware of the potential problems inherent in the use of COX-2 as well as COX-2 agents for chemoprevention in certain cancers. PMID- 12629932 TI - Use of the Pleurx Pleural Catheter for the management of malignant pleural effusions. AB - Many patients with cancer, especially those with lung and breast cancer, experience malignant pleural effusions. Several treatment options exist, and most require hospitalization. The Pleurx Pleural Catheter (Denver Biomedical, Golden, CO) is a new treatment option that allows patients to be treated on an outpatient basis for weeks or months. With a catheter in place, pleural effusions can be drained intermittently at home by trained family members or caregivers. Nurses play a critical role in educating patients about the use of the Pleurx catheter, as well as teaching patients and family members how to drain the catheter. The purpose of this article is to familiarize nurses with the proper care of Pleurx catheters and provide a basis for patient education. PMID- 12629933 TI - A comprehensive look at the early detection of ovarian cancer. AB - Ovarian cancer is the leading cause of gynecologic cancer deaths and the fifth leading cause of cancer deaths among American women. Advances in surgery and treatment modalities have slightly improved five-year survival rates, but the prognosis for most women continues to be poor. Rectovaginal examination, ultrasound, and the CA-125 blood test are three modalities currently used to screen for ovarian cancer, although no universal ovarian cancer screening guidelines are recommended for the general population. Developments in the early detection of ovarian cancer are emerging and include blood tests that could lead to identifying asymptomatic, early-stage ovarian cancer. Nurses should be knowledgeable about these developments in ovarian cancer as they educate women about ovarian cancer risk, prevention, and early detection. PMID- 12629934 TI - Treatment outcomes and quality-of-life issues for patients treated with prostate brachytherapy. AB - The increasing popularity of brachytherapy for treatment of early-stage prostate cancer requires oncology nurses to have a comprehensive knowledge of the disease, its treatment, and management of side effects. Because quality-of-life (QOL) issues have become an important consideration in treatment selection for many patients, oncology nurses must have a thorough understanding of these QOL issues and their management. Armed with knowledge about prostate brachytherapy and its effect on QOL, oncology nurses can offer accurate information and evidence-based symptom management techniques to patients undergoing brachytherapy for prostate cancer. PMID- 12629935 TI - Pegfilgrastim for chemotherapy-induced neutropenia. AB - Many chemotherapy regimens cause myelosuppression, which can result in febrile neutropenia and potentially lead to serious infections. The risk of neutropenia and its complications can be reduced with filgrastim, a granulocyte-colony stimulating factor. Filgrastim is safe and effective, but it is cleared rapidly from the body (predominantly through the kidneys) and requires daily administration for up to 14 days. A pegylated form of filgrastim, pegfilgrastim, has been developed by attaching a polyethylene glycol molecule to filgrastim. Pegfilgrastim has an extended circulation half-life and self-regulating, patient specific pharmacokinetics, making it possible to give the treatment as a single dose once per chemotherapy cycle. Clinical trials have shown that a single, subcutaneous dose of pegfilgrastim is as safe and effective as daily filgrastim injections in patients treated with myelosuppressive chemotherapy. In addition, a single, 6 mg fixed dose of pegfilgrastim per chemotherapy cycle is sufficient in adult patients, regardless of their body weight, making pegfilgrastim a simple, effective, and well-tolerated option for managing chemotherapy-induced neutropenia. PMID- 12629936 TI - Malignant hemangiopericytoma: a clinical overview and case study. AB - Although unusual, hemangiopericytoma (HPC) can be considered benign or malignant depending on histologic features. In some cases, malignant HPC can behave as high grade sarcomas. When HPC is deemed malignant, radical surgical excision is the treatment of choice. Adjuvant chemotherapy and radiation therapy offer limited success in patients with this malignancy. This article details a case study of a man with metastatic malignant HPC that spanned 25 months and three major cancer centers. Throughout the case study, chemotherapy, radiation therapy, and an experimental agent were employed to attempt to achieve control of the aggressive disease. Often, patients with HPC need a multidisciplinary approach to their care that requires involvement of nutritional and social services, behavioral medicine, and home or hospice care as appropriate. A better general understanding of this disease process is needed to meet the needs of patients with malignant HPC. PMID- 12629938 TI - Cardiotoxicity and capecitabine: a case report. AB - Oral fluoropyrimidines increasingly are being developed and studied as a novel treatment for breast, colorectal, and other cancers. Fluoropyrimidines are designed to generate 5-fluorouracil (5-FU) preferentially within tumors. Cardiotoxicity is a rare complication associated with 5-FU and oral fluoropyrimidine treatments. Chest pain is the most common presenting symptom, and, in many cases, the cardiotoxicity is partly or completely reversible. This article reviews fluoropyrimidine-induced cardiotoxicity and presents a case report of a woman who experienced this complication during capecitabine treatment. PMID- 12629937 TI - Developing and testing lay literature about breast cancer screening for African American women. AB - Written materials about breast cancer screening for African American women with low literacy skills are needed. Available materials were not at or below third grade reading levels, were not culturally sensitive, and were not accurate in illustrating correct breast self-examination (BSE) techniques. Focus groups representing the target population helped the authors design a pamphlet describing how to perform BSE and a motivational picture book to help women overcome barriers to screening. The authors chose a food theme for the cover of the pamphlet written at a third-grade level and suggested a photographic version. In the motivational book, two women address barriers to screening and replace myths and fears with facts and actions. Data from 162 women showed that they learned from both the photographic and illustrated versions. Women in the photographic group found significantly more lumps in the silicone models, so the authors chose that version to use in final testing. Finally, nurses pretested a group of patients before they reviewed the materials and post-tested another group after they reviewed them. The group who had reviewed the materials had greater knowledge of and intent to follow the guidelines and received higher scores on BSE techniques. PMID- 12629939 TI - In my house are many rooms: a proposed model to examine self-concept. PMID- 12629942 TI - Understanding non-Hodgkin's lymphomas. AB - NHLs are a complex group of diseases whose morphology and clinical presentation guide treatment choices. Nurses caring for these patients need to have an understanding about how these diseases present themselves and how the treatment options affect patients' emotional and physical well-being. PMID- 12629941 TI - What are the distinctions between Reiki and therapeutic touch? PMID- 12629940 TI - Thermal wounds following heating pad use. PMID- 12629943 TI - Evidence-based practice: making it happen in your clinical setting. PMID- 12629945 TI - Outpatient administration of radiolabeled monoclonal antibodies. PMID- 12629944 TI - Carboplatin dosing accounting for the renal and hematologic status of patients. AB - Using AUC-based dosing compensates for variations in renal function between and within individual patients. It does not adjust for other factors such as previous chemotherapy, previous radiotherapy, or performance status. One empiric method to adjust for these factors would be reduction of the ideal AUC. However, because AUC-based dosing accounts for variations in renal function, this would not be an appropriate method to adjust dosing based solely on renal function changes. PMID- 12629946 TI - Gelclair oral gel. PMID- 12629948 TI - Oncology nurses take time for rejuvenation and self-care. PMID- 12629947 TI - Ibritumomab tiuxetan. PMID- 12629949 TI - ONS takes appropriate steps to ensure sound financial health. PMID- 12629950 TI - Trauma therapy or exploitation? PMID- 12629951 TI - Obsessive-compulsive disorder. AB - TOPIC: Identification and management of obsessive-compulsive disorder (OCD). PURPOSE: To increase advanced practice clinicians' awareness of the prevalence, screening tools, diagnostic criteria, differential diagnosis, and therapeutic management of OCD. SOURCES: Published literature. CONCLUSIONS: Nurses have a pivotal role in teaching self-management techniques to people with OCD. PMID- 12629952 TI - Pharmacology for post-traumatic stress disorder related to childhood sexual abuse: a literature review. AB - TOPIC: Post-traumatic stress disorder (PTSD) related to pharmacotherapy responses in general and specifically to noncombat symptomatology related to childhood sexual abuse. PURPOSE: To explore findings related to pharmacological advances in the treatment of PTSD and relate the findings to the treatment of childhood sexual abuse. SOURCES: Published literature. CONCLUSIONS: The literature on pharmacotherapy for PTSD in general and childhood trauma and PTSD in particular is small and inconsistent, but we need to be aware of any and all advances. PMID- 12629953 TI - Mental health care providers' perception of giving culturally responsive care to American Indians. AB - PROBLEM: To determine if mental health care providers delivering culturally responsive care to American Indians experiencing severe and persistent mental illness. METHODS: This qualitative study used grounded theory. Eleven mental health care providers and two administrators participated on two American Indian reservations with two different tribal affiliations. FINDINGS: Preliminary findings indicate the ability of providers to be culturally responsive varied based on an awareness of their personal culture and of the diversity within the American Indian culture. CONCLUSIONS: Mental health providers need to develop a higher level of awareness of diversity within their culture, which is possible through greater connection to their own culture. Future research should ask clients what mental health services would be culturally responsive. PMID- 12629955 TI - Experimental psychology: quo vadis? PMID- 12629954 TI - Biological perspectives. Aripiprazole: a third generation of antipsychotics begins? AB - Schizophrenia places an incredible burden on patients, families, communities, and governments, therefore intense efforts to find therapeutic responses to this illness continue. The traditional antipsychotics were discovered by chance, the atypical antipsychotics were configured to preserve "what worked" while adding and eliminating important effects, and the next generation continues to evolve in an attempt to improve on previous generations. Aripiprazole is extoled as a new generation of antipsychotic drugs. Information to date suggests it is an effective antipsychotic with a remarkable side effect profile. We are hopeful this drug will provide a relief for the suffering associated with schizophrenia. PMID- 12629956 TI - Design-effects in prospective and retrospective memory for actions. AB - There is ample evidence that memory for action phrases such as "open the bottle" is better in subject-performed tasks (SPTs), i.e., if the participants perform the actions, than in verbal tasks (VTs), if they only read the phrases or listen to them. It is less clear whether also the sole intention to perform the actions later, i.e., a prospective memory task (PT), improves memory compared with VTs. Inconsistent findings have been reported for within-subjects and between-subjects designs. The present study attempts to clarify the situation. In three experiments, better recall for SPTs than for PTs and for PTs than for VTs were observed if mixed lists were used. If pure lists were used, there was a PT effect but no SPT over PT advantage. The findings were discussed from the perspective of item-specific and relational information. PMID- 12629957 TI - Age stereotypes in younger and older women: analyses of accommodative shifts with a sentence-priming task. AB - A sentence-priming technique was used to examine whether older women (N = 39) share a more positive view of aging than younger women (N = 35). Situationally specified statements about older and younger persons were presented, followed either by a semantically related word, an unrelated word of the same valence, or a nonword. The accessibility of target words was measured by reaction times in a lexical decision task. Whereas a semantic priming effect for negatively connoted materials emerged for both groups, a priming effect for positively connoted materials was found for older women only. Furthermore, an affective priming effect was found for the older group, i.e., older women tended to respond relatively faster (slower) to semantically unrelated positive (negative) words following a sentence about an older person. The results are discussed within a coping-theoretical framework. PMID- 12629958 TI - Remembering things that never occurred: the effects of to-be-forgotten stereotypical information. AB - Participants, 68 female and male nonpsychology university students, studied stereotypical and nonstereotypical words either with an initially activated social category (immigrant prime) or with no social category (neutral prime). They were then instructed to either forget or remember the studied words. This was followed by a recognition test. Based on the Deese-Roediger-McDermott (DRM) paradigm and stereotype-accessibility research, we anticipated that the participants in the immigrant, but not in the neutral, prime condition would falsely recognize more nonstudied stereotypic attributes under a forget than a remember instruction. The results supported our hypothesis. Implications of the outcome for eyewitness testimony, juridical decision-making, and stereotype maintenance are discussed. PMID- 12629959 TI - Implicit Association Test: separating transsituationally stable and variable components of attitudes toward gay men. AB - Implicit attitudes are conceived of as formed in childhood, suggesting extreme stability. At the same time, it has been shown that implicit attitudes are influenced by situational factors, suggesting variability by the moment. In the present article, using structural equation modeling, we decomposed implicit attitudes towards gay men into a person factor and a situational factor. The Implicit Association Test (Greenwald, McGhee, & Schwartz, 1998), introduced as an instrument with which individual differences in implicit attitudes can be measured, was used. Measurement was repeated after one week (Experiment 1) or immediately (Experiment 2). Explicit attitudes towards gay men as assessed by way of questionnaires were positive and stable across situations. Implicit attitudes were relatively negative instead. Internal consistency of the implicit attitude assessment was exemplary. However, the within-situation consistency was accompanied by considerable unexplained between-situation variability. Consequently, it may not be adequate to interpret an individual implicit attitude measured at a given point in time as a person-related, trait-like factor. PMID- 12629960 TI - Intuition beyond recognition: when less familiar events are liked more. AB - In a laboratory experiment, we compare the relative impact of two possible determinants of intuitive evaluative judgments: ease of recognition and total value of prior encounters with a target. Participants encode daily return values of shares on the stock market while watching videotaped ads on the computer screen. This dual-task procedure ensures that participants subsequently lack relevant event memories and thus have to rely on their intuition when evaluating the targets. In the presentation, the share appearing least frequently produced the highest sum of returns. In contrast, the share appearing most frequently produced the lowest sum of returns. Evaluative judgments reveal a preference for the share with the highest sum of returns, although, as evident from recognition latencies, it was the one that was more difficult to recognize. The results provide evidence for the value-account model of implicit attitude formation (Betsch, Plessner, Schwieren, & Gutig, 2001), which predicts that intuitive evaluative judgments reflect the total value of prior encounters. PMID- 12629961 TI - Relative influences of affect and cognition on behavior: are feelings or beliefs more related to blood donation intentions? AB - This study tested the relative predictive power of affect and cognition on global attitude and behavioral intention within the tripartite model of attitude structure. Participants (N = 264) completed questionnaires that included an item regarding blood donation experience, five semantic differential items, four behavioral intention items, and one global attitude item. Participants were randomly assigned to either an affective or cognitive instruction set for the semantic differential items. As predicted, semantic differentials were more highly correlated with both global attitude and behavioral intention when completed under the affective instructions than under the cognitive instructions. In addition, donors' and non-donors' attitudes on the semantic differential scales were distinguished from one another only when they were elicited under the affective instruction set. Results provide support for the tripartite model of attitude structure. Future research should examine the relative importance of affect and cognition in less emotion-laden domains. PMID- 12629962 TI - Strategic aspects of numerosity judgment: the effect of task characteristics. AB - In previous investigations we documented that people use several strategies to determine different numerosities of blocks that are presented in a square grid. One of these strategies is the clever subtraction strategy, wherein the number of empty squares in the grid is subtracted from the total number of squares in the grid. In the present study we investigated participants' flexibility in strategy use when varying the shape of the grid. Analysis of the results in terms of the theoretical framework of Lemaire and Siegler (1995) regarding strategic change shows that this contextual variable affected the frequency, execution time, and accuracy of subjects' use of the subtraction strategy. The usefulness of this framework for analyzing the nature of the adaptation to contextual variations is discussed. From a methodological point of view, this study documents the potential of Beem's (1993, 1999) segmented linear regression models for assessing subjects' strategy use in cognitive tasks. PMID- 12629963 TI - Advances in protein chemistry. Preface. PMID- 12629964 TI - Membrane protein assembly in vivo. PMID- 12629965 TI - Construction of helix-bundle membrane proteins. PMID- 12629966 TI - Transmembrane beta-barrel proteins. PMID- 12629967 TI - Length, time, and energy scales of photosystems. AB - The design of photosynthetic systems reflects the length scales of the fundamental physical processes. Energy transfer is rapid at the few angstrom scale and continues to be rapid even at the 50-A scale of the membrane thickness. Electron tunneling is nearly as rapid at the shortest distances, but becomes physiologically too slow well before 20 A. Diffusion, which starts out at a relatively slow nanosecond time scale, has the most modest slowing with distance and is physiologically competent at all biologically relevant distances. Proton transfer always operates on the shortest angstrom scale. The structural consequences of these distance dependencies are that energy transfer networks can extend over large, multisubunit and multicomplex distances and take leaps of 20 A before entering the domain of charge separating centers. Electron transfer systems are effectively limited to individual distances of 15 A or less and span the 50 A dimensions of the bioenergetic membrane by use of redox chains. Diffusion processes are generally used to cover the intercomplex electron transfer distances of 50 A and greater and tend to compensate for the lack of directionality by restricting the diffusional space to the membrane or the membrane surface, and by multiplying the diffusing species through the use of pools. Proton transfer reactions act over distances larger than a few angstroms through the use of clusters or relays, which sometimes rely on water molecules and which may only be dynamically assembled. Proteins appear to place a premium on robustness of design, which is relatively easily achieved in the long-distance physical processes of energy transfer and electron tunneling. By placing cofactors close enough, the physical process is relatively rapid compared to decay processes. Thus suboptimal conditions such as cofactor orientation, energy level, or redox potential level can be tolerated and generally do not have to be finely tuned. The most fragile regions of design tend to come in areas of complex formation and catalysis involving proton management, where relatively small changes in distance or mutations can lead to a dramatic decrease in turnover, which may already be limiting the overall speed of energy conversion in these proteins. Light-activated systems also face a challenge to robust function from the ever-present dangers of high redox potential chemistry. This can turn the protein matrix and wandering oxygen molecules into unintentional redox partners, which in the case of PSII requires the frequent, costly replacement of protein subunits. PMID- 12629968 TI - Structural clues to the mechanism of ion pumping in bacteriorhodopsin. PMID- 12629969 TI - The structure of Wolinella succinogenes quinol: fumarate reductase and its relevance to the superfamily of succinate: quinone oxidoreductases. PMID- 12629970 TI - Structure and function of quinone binding membrane proteins. PMID- 12629971 TI - Prokaryotic mechanosensitive channels. PMID- 12629972 TI - The voltage sensor and the gate in ion channels. PMID- 12629973 TI - Rhodopsin structure, dynamics, and activation: a perspective from crystallography, site-directed spin labeling, sulfhydryl reactivity, and disulfide cross-linking. PMID- 12629974 TI - The glycerol facilitator GlpF its aquaporin family of channels, and their selectivity. PMID- 12629976 TI - Ninth and eleventh circuits address EMTALA stabilization requirement. PMID- 12629975 TI - Summary judgment granted in copayment dispute. Alres v. Harvard Pilgrim Health Care Inc. PMID- 12629977 TI - Tricks for ultrasound-guided resection of colorectal liver metastases. AB - Nowadays, tumor clearance for colorectal liver metastases can be achieved in most patients undergoing liver resection with survival benefits. Most merits of conservative and radical resections should probably be addressed to the extensive use of intraoperative ultrasound. Mostly for this reason, surgery can be still considered the treatment of choice. For this purpose, intraoperative ultrasound guidance technique should be a familiar instrument for the hepatic surgeon. The technical tricks needed to perform a real intraoperative ultrasound-guided hepatectomy in the management of liver metastases from colorectal cancer are described. PMID- 12629978 TI - Hepatic encephalopathy secondary to transtumoral portal-hepatic venous shunting. AB - Intrahepatic portal-systemic shunts causing hepatic encephalopathy are very rare. This is a case report of a patient with hepatic metastases of a pancreatic islet cell tumor that manifested with transtumoral shunts leading to hepatic encephalopathy. The diagnosis was confirmed with Doppler ultrasound and initially treated with selective transhepatic portal vein embolization followed by hepatic artery embolization, and eventually radiofrequency ablation of the largest metastases. Despite excellent short-term palliation, symptom recurrence necessitated liver resection, the results of which proved durable. A multidisciplinary treatment plan for the identification and management of potentially salvageable encephalopathy in similar patients is described. PMID- 12629979 TI - Factors predicting concurrent cholangiocarcinomas associated with hepatolithiasis. AB - BACKGROUND/AIMS: Hepatolithiasis is a risk factor for cholangiocarcinoma. However, it is difficult to detect early cholangiocarcinoma that occurs as a complication of hepatolithiasis. To identify the factors, which can be used for predicting cholangiocarcinomas in patients with hepatolithiasis, we compared the clinical characteristics of patients who had cholangiocarcinoma associated with hepatolithiasis with those of patients with hepatolithiasis only. METHODOLOGY: Forty patients with cholangiocarcinoma associated with hepatolithiasis (group HC) and 73 patients with hepatolithiasis only (group H) were randomly selected for this study. Mean tumor size was 6.1 +/- 2.4 cm in diameter. RESULTS: Patients of group HC were older (56.7 +/- 8.9 yr) than those of group H (49.2 +/- 12.9 yr) (p < 0.001). Weight loss was more frequent in group HC (51.5%) than in group H (5.5%) (p < 0.001) and serum alkaline phosphatase levels were higher in group HC (181 +/- 184 IU/L) than in group H (426 +/- 385 IU/L) (p < 0.001). The proportion of the patients who had hepatolithiasis in the right or both lobes of the liver was higher in group HC (72.5%) than in group H (50.6%) (p = 0.024). The optimal cutoff value of serum CEA level for cholangiocarcinoma detection was set at 4.2 ng/mL using ROC cure to give a sensitivity of 67.6% and a specificity of 90.5%. Group HC differed from group H because of its lower rates of both abdominal pain and cholangitis, longer duration of stone history, and lower serum albumin level. Factors that did not predict cholangiocarcinoma included sex ratio, white blood cell count, serum bilirubin level, and hepatic atrophy. CONCLUSIONS: Cholangiocarcinoma should be suspected in patients with hepatolithiasis, especially when, the patient is over 40 years old, has a long history of hepatolithiasis with weight loss, a higher level of serum alkaline phosphatase, a lower level of serum albumin, a serum carcinoembryonic antigen level exceeding 4.2 ng/mL, and hepatolithiasis that is located either in the right or both lobes of the liver. PMID- 12629981 TI - Lymphatic spread differs according to tumor location in extrahepatic bile duct cancer. AB - BACKGROUND/AIMS: Identification of nodal involvement according to primary tumor location in extrahepatic bile duct carcinoma may guide surgical therapy. METHODOLOGY: Pathologic data of 81 patients who underwent curative operation for bile duct carcinoma were studied to clarify the differences in lymphatic spread from distal bile duct carcinoma, middle bile duct carcinoma, and proximal bile duct carcinoma. RESULTS: Lymph node metastases were present in 25 of 41 patients (61%) with distal bile duct carcinoma, 9 of 19 (47%) with middle bile duct carcinoma, and 11 of 21 (52%) with proximal bile duct carcinoma. The number of positive nodes per node-positive patient was greater in patients with middle bile duct carcinoma than in those with distal- or proximal bile duct carcinoma (mean 5.33 vs. 3.56 or 2.64, p < 0.05). Lymph nodes in the hepatoduodenal ligament were most frequently involved regardless of the primary tumor location. The frequency of distal- and middle bile duct carcinoma patients with metastasis to the superior mesenteric or para-aortic nodes was significantly higher than that of proximal bile duct carcinoma patients (p < 0.05 and p < 0.05). CONCLUSIONS: Patterns of lymphatic spread were different according to primary tumor location in bile duct carcinoma. Metastatic nodes were spread widely, from the hepatoduodenal ligament or posterior pancreaticoduodenal region to the nodes around the superior mesenteric artery and abdominal aorta, in distal- and middle bile duct carcinoma. PMID- 12629980 TI - The clinicopathological studies on patients with pancreaticobiliary maljunction that was detected by intraoperative cholangiography under laparoscopic cholecystectomy. AB - BACKGROUND/AIMS: The objective of this study was to evaluate the clinicopathological features of patients with pancreaticobiliary maljunction, which was detected by intraoperative cholangiography in laparoscopic cholecystectomy. METHODOLOGY: Among 78 patients who underwent intraoperative cholangiography in laparoscopic cholecystectomy, 8 patients had the radiological findings of pancreaticobiliary maljunction. The clinicopathological factors were analyzed in them. RESULTS: They consisted of 2 males and 6 females with a mean age of 57.8 years (range: 36-76 years). Preoperatively, 7 patients were diagnosed as cholelithiasis and one was diagnosed as polypoid lesions of the gallbladder. None of them had dilated choledochus. All of them underwent cholecystectomy alone extrahepatic bile duct resection was omitted, and was performed. Histopathologic examinations of the resected gallbladder revealed epithelial hyperplasia in one patient, in whom amylase level of the gallbladder bile was also highly elevated. Postoperatively, none of them had acute pancreatitis, cholangitis, or biliary cancer. CONCLUSIONS: Further prolonged careful follow-up is warranted in our series of patients. PMID- 12629982 TI - Mass-forming intrahepatic cholangiocarcinoma presenting tumor growth in extrahepatic bile duct and superficial spreading. AB - We present here a very unusual case in Western countries of mass-forming intrahepatic cholangiocarcinoma in association with tumor growth in the extrahepatic bile duct and superficial spreading. We discuss the clinicopathologic features of this very rare variant of cholangiocarcinoma. PMID- 12629983 TI - CT findings of cystic duct carcinoma two years before obstructive jaundice. AB - We describe a case of cystic duct carcinoma, the computed tomography findings of which had been incidentally obtained several times over the preceding three years before obstructive jaundice appeared. Under the diagnosis of extrahepatic bile duct carcinoma, a 75-year-old man underwent surgery. Two years before obstructive jaundice appeared, the arterio-portal phase of enhanced computed tomography demonstrated an ovoid mass with peripheral rim enhancement, measuring 1.5 cm in diameter, beside the extrahepatic bile duct. One year later, a repeat enhanced computed tomography consistently showed the ovoid mass with peripheral rim enhancement, and the lower slice of the contiguous computed tomography scan revealed a uniformly enhanced mass compressing the neck of the gallbladder. Four months before obstructive jaundice, the uniformly enhanced mass beside the extrahepatic bile duct markedly infiltrated the surrounding tissues with spicula formation. At laparotomy, a solid and hard tumor was localized on the right side of the extrahepatic bile duct, in the cholecystic duct and the neck of the gallbladder. Of 29 reported cases fulfilling Farrar's criteria in the English literature other than autopsy cases, computed tomography imaging was only performed in two cases, and the computed tomography findings of cystic duct carcinoma in its early stages, have never been fully described. PMID- 12629984 TI - Integrin alpha v, c-erbB2 and DNA ploidy in lung metastases from colorectal cancer. AB - BACKGROUND/AIMS: We analyzed integrin alpha v and c-erbB2 expression and DNA ploidy in primary tumors and pulmonary metastases from colorectal carcinoma. METHODOLOGY: Fifty primary tumors and 21 lung metastases were examined for immunohistochemical detection of integrin alpha v, c-erbB2 and DNA ploidy. RESULTS: In integrin alpha v-positive tumors, integrin alpha v was strongly recognized in the cytoplasm of colorectal carcinoma. In total cases, in a multivariate model using logistic stepwise regression analysis, venous invasion was significantly and independently related to lung metastasis in patients who underwent curative resection. Integrin alpha v, c-erbB2 and DNA ploidy in primary tumors were not predictable for lung metastasis. On the other hand, in the 21 patients with lung metastases, integrin alpha v overexpression was identified in 52.4% (11/21) of primary tumors and in 85.7% (18/21) of lung metastases, respectively. The positive ratio was significantly higher in lung metastases than in primary tumors (p < 0.05). There was no significant difference between primary tumors and lung metastases in DNA ploidy or c-erbB2 expression. CONCLUSIONS: Integrin alpha v might be one of the important signals for formation of lung metastases from colorectal cancer as a late event. PMID- 12629985 TI - Differential expression of pulmonary nitric oxide synthase isoforms after intestinal ischemia-reperfusion. AB - BACKGROUND/AIMS: Nitric oxide has been implicated in both attenuating and aggravating ischemia-reperfusion injury in most organs. This study aimed to investigate the role of nitric oxide produced by the two principal isoforms of nitric oxide synthase in the lung during post-ischemic reperfusion of the intestine. METHODOLOGY: Rats were randomized into four groups of 6 animals: Group A: laparotomy and superior mesenteric artery dissection without occlusion and maintenance for 2 h (control group at 2 h). Group B: laparatomy and superior mesenteric artery occlusion for 30 min and reperfusion of the intestine for 2 h (ischemia-reperfusion group at 2 h). Group C: control animals at 6 h. Group D: ischemia-reperfusion animals at 6 h. Arterial blood pressure was monitored throughout the procedure. Animals were euthanazed at the end of the experiment, and lungs were harvested for histological assessment of injury and for immunohistochemical examination of nitric oxide synthase isoforms and nitrotyrosine. RESULTS: In all animals subjected to intestinal ischemia a period of systemic hypotension occurred immediately upon reperfusion. Histological evidence of lung injury was limited to those animals subjected to an intestinal reperfusion insult. Compared to control animals, pulmonary endothelial nitric oxide synthase expression was diminished at 2 h (p = 0.002), while expression of inducible nitric oxide synthase (p = 0.002) and nitrotyrosine (p = 0.02) was increased at 6 h. CONCLUSIONS: Following intestinal ischemia-reperfusion, early pulmonary damage is associated with decreased endothelial nitric oxide synthase expression in the lung. Expression of inducible nitric oxide synthase occurs during the later stages of reperfusion; this leads to overproduction of nitric oxide with consequent nitrosylation of protein tyrosine residues and thus aggravated pulmonary injury. PMID- 12629986 TI - The subcutaneous splenic transposition prevents liver injury induced by excessive portal pressure after massive hepatectomy. AB - BACKGROUND/AIMS: We have proposed that acute portal hypertension, reflecting sinusoidal shear stress, becomes a trigger of liver regeneration after partial hepatectomy. Moreover, excessive shear stress induces liver injury. We investigated whether the use of a portosystemic shunt can reduce surplus damage of the remnant liver by means of excessive shear stress after massive hepatectomy. METHODOLOGY: In this study, to determine whether and by what mechanism excessive shear stress induces liver injury, we used the rat model to investigate whether a subcutaneous splenic transposition, which consists of a portosystemic shunt up to 4 weeks post-surgery, can prevent liver injury. RESULTS: Subcutaneous splenic transposition decreased the portal pressure immediately after 90% massive hepatectomy and relieved the elevation of transaminase and serum hyaluronic acid compared with findings of the no shunt group. In addition, the degree of leukocytopenia and thrombocytopenia in 90% massive hepatectomy with subcutaneous splenic transposition were better than those in 90% massive hepatectomy without subcutaneous splenic transposition. Serum tumor necrosis factor-alpha levels of the shunt group were lower than those of massive hepatectomy without subcutaneous splenic transposition. The rats that underwent 95% massive hepatectomy died within 48 hrs. The extent to which subcutaneous splenic transposition prolonged survival after 95% massive hepatectomy was statistically significant (mean survival of shunt group 60.9 +/- 13.4 hours vs. no shunt group 25.4 +/- 2.3 hours P = 0.001). CONCLUSIONS: These findings suggest that excessive shear stress after massive hepatectomy induces the liver injury against the hepatocytes and the sinusodial endothelial cells via intrahepatic microcirculation failure accompanied by overimmunoreaction. PMID- 12629987 TI - Free oxygen radical-induced acute pancreatitis. A light and electron microscopic study. AB - BACKGROUND/AIMS: To date direct toxic effects of free oxygen radicals in vivo on pancreatic parenchyma have not been studied thoroughly. We aimed to study: 1) the detailed histopathological changes induced by free oxygen radicals in pancreas; and 2) the preventive effect of intraductal catalase in H2O2-induced acute pancreatitis. METHODOLOGY: Wistar Albine rats were randomized into six groups. 1) First experiment: Bile-pancreatic duct was cannulated close to the liver and perfused through the duodenum with (i) normal saline solution, (ii) iron sulfate (FeSO4), (iii) hydrogen peroxide (H2O2), (iv) hydrogen peroxide and iron sulfate simultaneously. 2) Second experiment: Bile pancreatic duct was perfused either with H2O2 or H2O2 + catalase. Serum amylase and pancreas malondialdehyde levels were measured in both experiments after 3 hours of perfusion period. Tissue samples were obtained for histopathological examinations. RESULTS: 1) First experiment: Intraductal perfusion of FeSO4 or H2O2 or H2O2 + FeSO4 induced acute edematous pancreatitis with focal parenchymal necrosis. At the ultrastructural level, intracytoplasmic formation of vacuoles. fusion of the vacuoles and zymogen granules, and autophagosomes containing cellular organelles were found. Serum amylase and pancreas malondialdehyde levels, and morphological score were significantly higher in these groups than control group (p < 0.001). 2) Second experiment: Catalase perfusion simultaneously with H2O2 decreased the serum amylase and pancreas malondialdehyde levels, and morphological score significantly (p < 0.001) and prevented the desquamation of the columnar epithelium and development of gross edema but not parenchymal necrosis. CONCLUSIONS: Intraductal perfusion of FeSO4 or H2O2 or H2O2 + FeSO4-induced acute pancreatitis with marked light and electronmicroscopic changes. Intraductal perfusion of catalase and H2O2 simultaneously did not prevent or lessen the parenchymal necrosis. These findings have suggested that another mechanism of injury may also play a role in parenchymal injury in oxygen radical-induced acute pancreatitis. PMID- 12629988 TI - Iatrogenic gastroduodenal ulcers during hepatic intra-arterial chemotherapy. AB - BACKGROUND/AIMS: Various percentages of iatrogenic gastroduodenal ulcers during hepatic intra-arterial chemotherapy have been reported in the literature. The aim of this study was to analyze a homogeneous cohort of patients in order to evaluate the evolution and management of this complication. METHODOLOGY: We retrospectively reviewed the clinical charts of 80 patients with primary or metastatic liver tumors who received 186 hepatic arterial infusion chemotherapy courses of 5-fluorouracil, cisplatin and mitomycin-C. All of the patients complaining of upper gastrointestinal symptoms during or after hepatic arterial infusion underwent esophagogastroduodenoscopy. RESULTS: Esophagogastroduodenoscopy was performed in 14 patients, all of whom had gastroduodenal ulcers. Two of ten investigated patients were Helicobacter pylori positive. All of the patients were treated with a proton pump inhibitor and five also received major analgesics. All of the ulcers healed without complications. Six patients did not continue with hepatic arterial infusion for reasons other than ulcers. Eight patients received a subsequent hepatic intra-arterial chemotherapy course, five despite the persistence of an active ulcer. CONCLUSIONS: Iatrogenic gastroduodenal ulcers are probably due to ischemia and the direct toxicity of the anticancer agents. They are Helicobacter pylori independent and do not represent an absolute contraindication for the continuation of hepatic intra-arterial chemotherapy. PMID- 12629989 TI - Expression of transforming growth factor-beta 1 in hepatocellular carcinoma in comparison with the non-tumor tissue. AB - BACKGROUND/AIMS: TGF-beta 1 (transforming growth factor-beta 1) has been shown to be overexpressed in various cancer cells including hepatocellular carcinoma cells. In this study, we examined the association of the immunohistochemical expression of TGF-beta 1 in hepatocellular carcinoma tissues with the clinicopathological findings. METHODOLOGY: Thirty liver tumor biopsy specimens obtained with ultrasound guidance and in which tumor-tissue and non-tumor tissue existed were examined. TGF-beta 1 was stained using rabbit anti-human TGF-beta 1 antibody. RESULTS: TGF-beta 1 was expressed mainly in the cytoplasm and plasma membrane of the hepatocellular carcinoma cells, while it was stained in the fibrous septa and sinusoidal cells in the non-tumor tissue. When comparing the intensity of TGF-beta 1 staining in the hepatocellular carcinoma tissue with that in the non-tumor tissue, the former was more intense than the latter in 21 (70%) of the 30 cases. Although the expression of TGF-beta 1 in the hepatocellular carcinoma tissue was not related to the tumor size of hepatocellular carcinoma, it was correlated with the histological differentiation: the lower the histological differentiation grade of hepatocellular carcinoma was, the more intense was the TGF-beta 1 staining in hepatocellular carcinoma tissue. In addition, TGF-beta 1 tended to be overexpressed in the trabecular type of hepatocellular carcinoma. To elucidate the relationship of TGF-beta 1 staining with apoptosis in hepatocellular carcinoma cells, we performed terminal deoxynucleotidyl transferase-mediated DUTP biotinnick end labeling (TUNEL) in 11 of the 30 specimens. No positive cells for TUNEL were detected in the hepatocellular carcinoma tissues. CONCLUSIONS: These results indicated that the expression of TGF-beta 1 was associated with the histological differentiation of hepatocellular carcinoma cells. PMID- 12629990 TI - Sinistroposition of the gallbladder and the common bile duct. AB - A left lateral bisegmentectomy was performed in a 29-year-old man presenting a primary lymphoma of the liver. Surgical exploration revealed a left-side gallbladder, located under the left lobe of the liver. During hepatic parenchyma dissection, performed strictly at the left of the round ligament and the umbilical portion of the left portal vein, common bile duct was injured. Complete separation of hepatic pedicle structures showed that the upper biliary convergence passed on the left side of the umbilical portion of the left portal vein before reaching the hepatoduodenal ligament. This case report discusses the embryological mechanism that could explain this uncommon bile duct abnormality, focusing on its consequences during left ruled lobectomy. PMID- 12629991 TI - Treatment of gastrojejunal anastomotic leak with a covered metallic stent. AB - Although the incidence rates of anastomotic leaks after radical gastrectomy in a patient with gastric cancer are generally low, when the complication ensues, morbidity and mortality rates rise, varying between 18% and 30%. We used self expandable, covered metallic stent to occlude the leakage site in a patient with recurrent postoperative anastomotic leaks. The deployment of the covered stent was successful, and immediate leak occlusion was obtained after the procedure. During follow-up, the patient had unrecognized passage of the stent per rectum and showed no anastomotic leak on upper gastrointestinal series. We suggest that implementation of covered metallic stent should be considered as a valuable, alternative therapeutic option for the treatment of intractable postoperative anastomotic leak after radical gastrectomy. PMID- 12629992 TI - Occurrence of hepatocellular and cholangiocellular carcinoma in different hepatic lobes. AB - Separate hepatocellular and cholangiocellular carcinoma (double cancer) in the liver are extremely rare subtypes of primary hepatic carcinomas. We report a case of double primary liver carcinomas that were surgically resected simultaneously. A 66-year-old man was admitted because of elevation of serum levels of alpha fetoprotein. Abdominal computed tomography and angiography showed two hypervascular masses in S4 and S8 hepatic segments. With the diagnosis of multiple hepatocellular carcinomas, the tumors were surgically resected. Histological examination showed that the tumor in S4 segment was moderately differentiated cholangiocellular carcinoma, the other in S8 segment was trabecular, moderately differentiated hepatocellular carcinoma. Immunohistochemically, a positive staining in carcinoembyonic antigen and cytokeratin 7 supported the diagnosis of cholangiocellular carcinoma for the tumor in S4 segment. The frequency of double cancer in the liver is much lower than mixed or combined cancer (0.1-0.5%). The different epithelial malignant tumors of hepatocellular carcinoma and cholangiocellular carcinoma, which were located in different hepatic lobes and resected simultaneously, has been reported in only two cases including the present case. PMID- 12629994 TI - Primary pleomorphic rhabdomyosarcoma of the liver: a case report. AB - Primary intrahepatic rhabdomyosarcoma is extremely rare in children. We describe a case of pleomorphic rhabdomyosarcoma originating from the liver in an eight year-old boy presenting with abdominal pain, spiking fever and a rapidly growing abdominal mass for one week. Preoperative imaging studies revealed a large solid tumor in the right lobe of the liver without any tumor elsewhere in the body. Serological study was negative for HBsAg and positive for anti-HBs. Biochemical tests including serum glutamic-oxalacetic and glutamic-pyruvic transaminase, alkaline phosphatase, bilirubin and alpha-fetoprotein were all within normal limits. The tumor was removed by an extended right hepatectomy. Histological and immunohistochemical examination confirmed a pleomorphic rhabdomyosarcoma. The patient eventually succumbed to tumor recurrence with massive internal hemorrhage two months after resection of the tumor. This is the first report of primary pleomorphic rhabdomyosarcoma of the liver in children. Abdominal ultrasonography, computed tomography scan and angiography are valuable for preoperative planning but the imaging findings are non-specific. Pathological examination with immunohistochemical stains remains the most important method in arriving at the exact diagnosis. The poor prognosis and early death of most previously reported cases imply the need for investigation of a more effective treatment method of this uncommon tumor. PMID- 12629993 TI - Can ethanol therapies injure the bile ducts? AB - Administration of ethanol, whether applied directly to tissue or via an intra arterial route, has been widely used to treat a variety of hepatic disorders, most importantly hepatocellular carcinoma. Animal studies, however, have demonstrated that intravascular hepatic ethanol therapy causes significant bile duct injury, and therefore, many centers have used intravascular ethanol applications with caution. A case of extensive bile duct injury in a 35-year-old female with a symptomatic hepatic hemangioma treated with ethanol embolization is presented. Although a seemingly rare complication, the possibility of bile duct injury should be considered prior to planned ethanol liver treatment, and a high index of suspicion should be maintained should signs of hepatic dysfunction subsequently arise. PMID- 12629995 TI - Successful treatment of intractable esophagothoracic fistula using covered self expandable stent. AB - This report concerns the successful treatment with a covered self-expandable stent of an intractable thoracoesophageal fistula after total esophagectomy for esophageal cancer. Total esophagectomy was performed on a 68-year-old man who presented with a huge esophageal cancer in the lower esophagus. Massive leakage was observed on the 5th day postoperatively. Since high fever and coughing continued, he was diagnosed as having esophagothoracic fistula and pyothorax, after which fenestration of the right chest wall was performed. Although the patient's general condition was getting better, stenosis near the anastomosis (esophagogastrostomy) and the esophagothoracic fistula were resistant to treatment with balloon dilatation and repeated endoscopic mucotomy. Further treatment, consisting of glue or fibrin sealant injection was not effective. After a covered self-expandable stent had been placed endoscopically, however, the fistel was completely cured in 2 months. This new endoscopic approach thus represents a promising option for the treatment of intractable esophagothoracic fistula. PMID- 12629996 TI - Decreased production of anti-mouse antibody after administration of human/mouse chimeric monoclonal antibody-neocarzinostatin conjugate to human. AB - BACKGROUND/AIMS: One problem that has been encountered with the use of murine Mabs is the development of a human anti-mouse antibody (HAMA) which lead to serious problems such as anaphylaxis. In order to evaluate the decreased HAMA production after administration of Fab fragments of chimeric monoclonal antibody, Fab fragments of chimeric Mab A7 (chA7Fab) was administered as neocarzinostatin (NCS) conjugate to seven patients with colonic cancer. METHODOLOGY: The in vivo pharmacokinetics of chA7Fab and HAMA production was examined. These patients received an infusion of 4,000 U: dose of NCS (18 mg: dose of chA7Fab). RESULTS: The plasma disappearance curves showed that chA7Fab-NCS was more rapidly cleared than A7-NCS. The chA7Fab-NCS did not elicit of HAMA in two of seven evaluable patients. The chA7Fab-NCS NCS elicited low levels of HAMA in five of seven evaluable patients. In contrast, A7-NCS elicited high levels of HAMA in all patients tested. Anti-isotype HAMA was not seen in seven evaluable patients tested with chA7Fab-NCS, while A7-NCS elicited high levels in all patients tested. CONCLUSIONS: This study demonstrates the reduced immunogenicity and shorter clearance time of chA7Fab-NCS compared to A7-NCS. PMID- 12629997 TI - Colonic preparation correlates with fasting breath hydrogen in patients undergoing colonoscopy. AB - BACKGROUND/AIMS: Fasting breath hydrogen has been reported to be low after fasting and polyethylene glycol ingestion. However, the relationship between fasting breath hydrogen and colonic preparation for colonoscopy has not been studied before. We evaluated fasting breath hydrogen in patients undergoing colonoscopy and correlated it with the quality of colonic preparation. METHODOLOGY: One day before colonoscopy, the patients ingested clear liquid diet and starting in the afternoon the day before colonoscopy they drank polyethylene glycol 8oz every 10 minutes for 3 hours. They fasted overnight. Fasting breath hydrogen was measured using an EC 60 gastrolyzer. Colonic preparation was rated as excellent, fair or poor. The colonoscopic findings were noted. RESULTS: There were 69 male patients. 89% had excellent or fair colonic preparation (Group A) 11% had poor preparation (Group B). Fasting breath hydrogen in Group B patients 14.5 +/- 1.5 ppm was significantly higher compared to that in Group A patients 2.2 +/- 0.2 ppm (p < 0.005 t-test). In other respects the two groups were similar. CONCLUSIONS: Patients undergoing colonoscopy after polyethylene glycol ingestion who have poor colonic preparation have higher fasting breath hydrogen compared to those who have excellent/fair colonic preparation. The clinical implication of this observation is discussed. PMID- 12629998 TI - Utility of serosal stamp cytology as an indicator for high-risk peritoneal metastasis in colorectal cancer surgery. AB - BACKGROUND/AIMS: Although peritoneal lavage cytology is widely performed during surgery for gastric cancer and the results have been reported to provide an accurate prognostic factor, its value has not been well established in colorectal cancer. In this study, we demonstrated the utility of serosal stamp cytology from the viewpoint of cell adhesion molecules. METHODOLOGY: Between 1997 and 1999, peritoneal lavage cytology and serosal stamp cytology were performed in 34 patients with resectable colorectal cancer. Epithelial cadherin (E-cadherin) was examined as an index of the progress degree of peritoneal metastasis. RESULTS: Although peritoneal lavage cytology was positive in one case, serosal stamp cytology was positive in 10 cases. E-cadherin expression was lost in all peritoneal lavage cytology and/or serosal stamp cytology positive patients. CONCLUSIONS: Our data indicate that serosal stamp cytology is more sensitive and simple than peritoneal lavage cytology. Serosal stamp cytology may be useful in identifying patients at high risk for peritoneal recurrence. PMID- 12629999 TI - Efficacy of cyclosporin with corticotropin for refractory ulcerative colitis. AB - BACKGROUND/AIMS: Cyclosporin was reported to be useful for steroid-resistant severe ulcerative colitis in the short term, but limited data are available on the long-term follow-up of such patients. Our aim was to assess the short- and long-term efficacy of combination therapy with cyclosporin and corticotropin for steroid-resistant ulcerative colitis. METHODOLOGY: Twenty-one patients with ulcerative colitis who did not respond to corticosteroid therapy, were treated with corticotropin, and 9 patients (43%) of them achieved complete remission. Twelve patients (57%) who did not achieve complete remission by corticotropin alone were given combination therapy with cyclosporin and corticotropin. RESULTS: In 12 patients who received combined therapy with cyclosporin and corticotropin, clinical severity was distinctly improved in 11 patients (92%) by combination therapy within 2 weeks and 7 patients (58.3%) entered into complete remission with salicylazosulfapyridine or 5-aminosalicylic acid alone. Two patients (16.7%) demonstrated insufficient effect and continue to receive a lower dosage of cyclosporin or corticosteroid. Three patients (25%) failed to respond to the combination therapy and required colectomy. Three of 7 patients who entered into remission relapsed 0.5, 5 and 5.5 months (average: 3.7 months) after cyclosporin withdrawal, but the clinical severity at the time of relapse was milder than that at the beginning of the treatment, namely, moderate in 2 patients, and mild in 1 patient. There were no significant adverse effects in our series. CONCLUSIONS: We demonstrated that oral cyclosporin in combination with corticotropin was highly effective for ulcerative colitis refractory to corticosteroid or corticotropin therapy and severe relapse was uncommon during several years of follow-up. PMID- 12630000 TI - Intussusception up to splenic flexure by cecal carcinoma in an adult: report of a case. AB - A 73-year-old woman was admitted because of constipation and appetite loss. She was diagnosed as having intussusception caused by a colonic tumor, based on the results of physical examination and imaging such as ultrasonography, computed tomography and barium enema. Operation revealed that right colon from the cecum up to the hepatic flexure of the ascending colon was not fixed to the retroperitoneum, and a circular cecal carcinoma was invaginated to the splenic flexure of the transverse colon. We experienced a rare case of ileocolic intussusception up to the splenic flexure by a cecal carcinoma with mesenterium ileo-colicum commune in an adult. PMID- 12630001 TI - Resection of a pelvic schwannoma with partial removal of the sacral nerve root. AB - Because the symptoms do not appear in most cases until the tumor has grown quite large, schwannomas are often detected incidentally during work-up for some other condition. Benign schwannomas generally do not invade adjacent structures but making the diagnosis preoperatively can be difficult when the tumor is large. We report the case of a benign schwannoma which was diagnosed preoperatively by needle core biopsy under computed tomography guidance. The tumor was removed completely by partially resecting the S1 nerve root, which was subsequently reanastomosed. The postoperative course was uneventful with the exception of minimal left leg pain due to neurologic deficit. PMID- 12630002 TI - The influence of endoscopic variceal ligation on the portal pressure gradient in cirrhotics. AB - BACKGROUND/AIMS: After variceal eradication by endoscopic ligation, fundal varices and worsening of portal hypertensive gastropathy can occur. The aim of this study is to verify the impact of the eradication of esophageal varices by endoscopic ligation on the portal pressure gradient, worsening of portal hypertensive gastropathy and development of fundal varices. METHODOLOGY: Twenty two (15M/7F, mean age: 54.5 years) cirrhotics with previous variceal bleeding were submitted to measurement of hepatic venous pressure gradient before and after variceal eradication by endoscopic ligation. RESULTS: The mean hepatic venous pressure gradient in the first measurement was 14.1 mmHg and after eradication, 13.5 mmHg (p = 0.403). After eradication, 12 patients experienced a reduction in portal pressure and 10, an elevation. Three patients developed fundal varices. Their mean gradient before treatment was 22 mmHg and 18.8 mmHg after therapy (p = 0.368). The gastropathy worsened in 9 patients (mean gradient before therapy of 15.2 mmHg; and 16.1 mmHg after treatment) (p = 0.303). The initial pressure gradient of these patients was not different from the other 13 cases (p = 0.463). CONCLUSIONS: The esophageal variceal eradication by endoscopic band ligation does not alter the hepatic venous pressure gradient. There is no significant variation in the portal pressure of patients in whom there was a worsening of portal hypertensive gastropathy or fundal varices development. PMID- 12630003 TI - Using non-invasive radionuclide imaging to detect esophagitis in patients with gastroesophageal reflux disease. AB - BACKGROUND/AIMS: The aim of our study was to evaluate technetium-99m tetrofosmin SPECT (single photon emission computed tomography) of the chest as a potential non-invasive method to evaluate esophagitis among gastroesophageal reflux disease patients. METHODOLOGY: In this study, we used non-invasive radionuclide imaging technetium-99m tetrofosmin chest SPECT to detect and grade esophagitis in 60 patients with gastroesophageal reflux disease. RESULTS: Based on the endoscopic findings according to the Savary-Miller system, the sensitivity, specificity, and accuracy rates of technetium-99m tetrofosmin chest SPECT in detecting esophagitis were 95.2%, 72.2%, and 88.3%, respectively. The agreement between the endoscopic and technetium-99m tetrofosmin chest SPECT findings was evaluated and found to be good (kappa: 0.633, p < 0.001). CONCLUSIONS: Non-invasive technetium-99m tetrofosmin chest SPECT has excellent sensitivity and good accuracy as a screen for esophagitis among gastroesophageal reflux disease patients. PMID- 12630004 TI - Physiological studies on nitric oxide in the lower esophageal sphincter of patients with reflux esophagitis. AB - BACKGROUND/AIMS: Nitric oxide has recently been shown to be a neurotransmitter in the non-adrenergic non-cholinergic inhibitory nerves in the digestive tract. To clarify the significance of nitric oxide in the lower esophageal sphincter of patients with reflux esophagitis, we have investigated enteric nerve responses in lower esophageal sphincter specimens obtained from the patients with gastric cancer who had reflux esophagitis, using the normal lower esophageal sphincter as a control. METHODOLOGY: Lower esophageal sphincter specimens were obtained from 6 patients who had gastric cancer with reflux esophagitis, and normal lower esophageal sphincter specimens were obtained from 12 patients who had gastric cancer without gastroesophageal reflux disease. A mechanograph was used to evaluate in vitro lower esophageal sphincter muscle responses to electrical field stimulation of the adrenergic and cholinergic nerves before and after treatment with various autonomic nerve blockers, and NG-nitro-L-arginine and L-arginine. RESULTS: 1) Cholinergic nerves were more dominant in the normal lower esophageal sphincter than in the lower esophageal sphincter with reflux esophagitis (p < 0.01); 2) non-adrenergic non-cholinergic inhibitory nerves were significantly found to act more on the lower esophageal sphincter with reflux esophagitis than those in the normal lower esophageal sphincter (p < 0.01); 3) nitric oxide mediates the relaxation reaction via non-adrenergic non-cholinergic inhibitory nerves in the normal lower esophageal sphincter and the lower esophageal sphincter with reflux esophagitis; 4) The relaxation reaction of nitric oxide was observed in the normal lower esophageal sphincter and increased in the lower esophageal sphincter with reflux esophagitis. CONCLUSIONS: These findings suggest that the cholinergic and non-adrenergic non-cholinergic inhibitory nerves play important roles in regulating contraction and relaxation of the human lower esophageal sphincter, and nitric oxide plays an important role in non-adrenergic non-cholinergic inhibitory nerves of the human lower esophageal sphincter. In addition, a decrease of the action of cholinergic nerves and an increase of the action of non-adrenergic non-cholinergic inhibitory nerves by nitric oxide may be largely related to the low lower esophageal sphincter pressure observed in the patients with reflux esophagitis. PMID- 12630005 TI - Pattern of recurrence after extended esophagectomy for squamous cell carcinoma of the esophagus. AB - BACKGROUND/AIMS: To further investigate the underlying mechanism of the systemic spread of esophageal squamous cell carcinoma. METHODOLOGY: Out of 151 patients who underwent a curative esophageal resection, 41 (27.1%) developed recurrent esophageal cancer. Nine recurrences (22%) were distant-hematogenous, 17 (41.5%) non-hematogenous, and 15 (36.5%) mixed. Hematogenous deposits accompanied 58.5% of the recurrences. The relation between several clinicopathological factors and the pattern of recurrence was evaluated. RESULTS: Univariate analysis recognized the lack of adjuvant chemoradiation, the tumor location in the lower esophagus and the tumor dedifferentiation as promoting factors for hematogenous recurrence. Poorly differentiated or undifferentiated tumors presented a significantly higher microvessel density than moderately or well differentiated tumors. Tumor differentiation and tumor lower localization were independent predictors of hematogenous recurrence. CONCLUSIONS: Patients with poorly differentiated or undifferentiated tumors, which are located at the lower esophagus and present high microvessel density, should be considered at high risk for hematogenous recurrences after extended esophagectomy. PMID- 12630006 TI - Questionnaire for early detection of occult anemia. AB - BACKGROUND/AIMS: Anemia is a common condition that is often missed. We aimed to devise a questionnaire for detection of occult anemia. METHODOLOGY: A questionnaire survey was done of 442 new gastroenterology outpatients to detect potential symptoms of anemia. Then hematology tests were performed and anemia was defined by World Health Organization criteria. Data were analyzed by the stepwise logistic regression method to determine adjusted odds ratios and extract relevant items. After further selection of variables, the sensitivity and specificity of three potential questionnaires were analyzed using receiver operating characteristic curves. RESULTS: Seven items were selected and a new questionnaire was devised to detect occult anemia. It showed a sensitivity of 70% and also a specificity of 70% when back-tested on the subjects. CONCLUSIONS: This new questionnaire may be useful for detecting occult anemia. PMID- 12630007 TI - Antioxidant potential in esophageal, stomach and colorectal cancers. AB - BACKGROUND/AIMS: The gastrointestinal tract is particularly susceptible to reactive oxygen species attack which lead to carcinogenesis. An important role in defense strategy against reactive oxygen species is played by antioxidants. The present study aims at examining antioxidant parameters and malondialdehyde--the product of lipid peroxidation as well as the marker of cancer progression--and cancer procoagulant in esophageal, gastric and colorectal cancer patients. METHODOLOGY: The activity of superoxide dismutase, catalase, glutathione peroxidase and reductase and the level of glutathione, vitamin C, malondialdehyde and cancer procoagulant were determined in tumors and normal mucous from 18 patients with esophageal cancer, 18 patients with stomach tumor and 62 patients with colorectal cancer. RESULTS: In esophageal tumor the activity of all enzymes has been increased compared with normal mucous. Stomach tumor has been also characterized by an increase in antioxidant enzymes activity except glutathione peroxidase and reductase whose activities have been decreased. However in colorectal tumor the activity of enzymes has been increased apart from catalase. In all cases the glutathione level has been increased while the vitamin C content has been significantly decreased. Tumor malondialdehyde level was significantly increased, too. The level of cancer procoagulant also increased in cancer tissues as well as in the serum. CONCLUSIONS: Antioxidant potential in all cases of gastrointestinal tract cancer has been unbalanced which has lead to increase in reactive oxygen species action and enhancement of lipid peroxidation and cancer procoagulant generation. PMID- 12630008 TI - Thrombopoietin serum levels in patients with inflammatory bowel disease with and without previous thromboembolic events. AB - BACKGROUND/AIMS: Patients affected by inflammatory bowel disease frequently suffer from thromboembolic complications and mesenteric microvascular occlusion could be involved in the pathogenesis of inflammatory bowel disease. Increased platelet counts and abnormal platelet function seem to play a crucial role in determining the hypercoagulable state observed in inflammatory bowel disease. Thrombopoietin is considered the primary regulator of thrombopoiesis and recent studies have investigated the role of thrombopoietin in inflammatory bowel disease. However, the available data are not conclusive. The aim of this study was to assess thrombopoietin serum levels in inflammatory bowel disease patients according to platelet counts, disease activity and previous thrombotic events. METHODOLOGY: Seventy-one patients with inflammatory bowel disease [41 with ulcerative colitis and 30 with Crohn's disease] and 30 healthy controls were investigated. Eight (11%) inflammatory bowel disease patients had suffered previous thromboembolic complications, none had active thrombosis. Thrombopoietin serum levels were measured by ELISA. RESULTS: Mean thrombopoietin levels were significantly increased in inflammatory bowel disease patients with active disease compared to both healthy controls and patients with inactive disease. Platelet counts were significantly higher only in patients with active disease with respect to healthy subjects. No correlation was found between thrombopoietin levels and platelet counts in either controls or inflammatory bowel disease patients. No differences were found either in thrombopoietin levels or in platelet counts comparing inflammatory bowel disease patients with and without thromboembolic complications. CONCLUSIONS: Our data show elevated thrombopoietin levels in active inflammatory bowel disease. However, no correlation was found between platelet counts and thrombopoietin levels, supporting the hypothesis that other circulating factors than thrombopoietin interact in determining reactive thrombocytosis. Furthermore, thrombopoietin levels did not differ in inflammatory bowel disease patients with or without previous thromboembolic events. This finding could be probably explained by the lack of patients with active thrombosis at the moment of inclusion in the study. PMID- 12630009 TI - Comparison between technetium-99m hexamethylpropyleneamineoxide labeled white blood cell abdomen scan and abdominal sonography to detect appendicitis in female patients with an atypical clinical presentation. AB - BACKGROUND/AIMS: Acute appendicitis is a clinically challenging surgical disease which is particularly difficult to diagnosis in women. An atypical presentation is a major obstacle to quickly diagnose acute appendicitis. METHODOLOGY: Fifty female patients with acute abdomen pain who were suspected of having appendicitis, but who had atypical findings were included in this study. After an intravenous injection of technetium-99m hexamethylpropyleneamineoxide (Tc-99m HMPAO) labeled white blood cells, serial anterior abdomen-pelvis images at 30 min, 60 min, 120 min and 240 min were obtained using a gamma camera. Meanwhile, at the point of maximal tenderness, abdominal sonography was performed with a graded compression technique for both longitudinal and transverse images. RESULTS: Thirty-two patients received a laparotomy for surgical and pathological diagnoses. The remaining 18 patients did not receive operations and showed no evidence of appendicitis after at least a one-month follow-up period. Two patients had false-positive white blood cell scan findings and 2 patients had false-negative white blood cell scan findings. The overall sensitivity, specificity, and accuracy for white blood cell scans in diagnosing appendicitis was 93.3%, 90.0%, 92.0%, respectively. One patient had false-positive abdominal sonographic finding and 5 patients had false-negative abdominal sonographic findings. The overall sensitivity, specificity, and accuracy for abdominal sonography scans in diagnosing appendicitis were 83.3%, 95.0%, and 88.0%, respectively. CONCLUSIONS: The Tc-99m HMPAO white blood cell scan provides a more sensitive and accurate method for the diagnosis of appendicitis in female patients with equivocal clinical presentation when compared with abdominal sonography. PMID- 12630010 TI - Portal embolization in various types of liver: novel variables to predict hypertrophy. AB - BACKGROUND/AIMS: The aim of this study was to investigate the hypertrophic effect of portal embolization in various types of liver and clarify useful variables, for predicting efficacy of portal embolization. METHODOLOGY: Portal embolization was performed for 46 patients with hepatocellular carcinoma (n = 30), biliary tract cancer (n = 9), or metastatic liver tumors (n = 7). The hypertrophic effect of portal embolization in relation to diseases, clinical liver conditions, histological fibrosis, and liver function were examined. RESULTS: The hypertrophic effect of portal embolization was impaired in the patients with hepatocellular carcinoma, chronic hepatitis/cirrhotic liver, and advanced liver fibrosis. ICGR15 (indocyanine green dye retention rate at 15 minutes) was revealed to be an independent adverse predicting factor. Especially in hepatocellular carcinoma patients, platelet count was significantly correlated with the hypertrophy ratio. In patients who underwent major hepatectomy for hepatocellular carcinoma, not only the incidences of posthepatectomy liver failure but also survival rate were similar between patients with and without portal embolization, although patients with portal embolization originally had a limited liver function. CONCLUSIONS: Preoperative portal embolization made major hepatectomy possible in hepatocellular carcinoma patients, although portal embolization was less effective compared with other diseases. ICGR15 and platelet count may be novel variables to predict the hypertrophic effect of portal embolization in all and hepatocellular carcinoma patients, respectively. PMID- 12630011 TI - Retinyl-palmitate reduces liver fibrosis induced by biliary obstruction in rats. AB - BACKGROUND/AIMS: The administration of retinoid in various experimental models of induced hepatic cirrhosis has shown antifibrogenic effects. The purpose of this study was to verify if vitamin A could interfere in hepatic fibrosis induced by biliary obstruction. METHODOLOGY: Male Wistar rats that underwent biliary obstruction were treated 2 weeks before the surgery with a dose of 50 x 10(3) IU retinyl-palmitate, twice a week, and then given the dosage for 5 weeks following surgery. They were then sacrificed and hepatic fragments removed to find out the biochemical dosage of hydroxyproline and the stereologic volume of collagen. Blood samples were also collected to find the biochemical dosage. RESULTS: The rats that underwent biliary obstruction and received vitamin A presented lower levels of hepatic hydroxyproline (p < 0.001) as well as a lower percentage of collagen tissue than the untreated control rats, but they presented higher AST and ALT serum levels (p < 0.05, p < 0.05). CONCLUSIONS: Under the above experimental conditions, the administration of vitamin A significantly reduced hepatic fibrosis. PMID- 12630012 TI - Molecular evidence for transmission of GB virus-C/Hepatitis G virus infection within family: close relationship between mother and child. AB - BACKGROUND/AIMS: GBV-C/Hepatitis G virus (HGV) is transmitted parentally, but some of the transmitted routes are still unclear. This study was conducted to investigate the relationship of family members in the transmission of HGV. METHODOLOGY: The study group enrolled 11 children from four index mothers with HGV viremia, whereas the control group had 14 children from seven matched mothers without HGV infection. Detection of circulating HGV RNA and anti-E2 antibody was conducted. Comparison of partially HGV nucleotide sequences at the 5' non-coding and non-structure 5 regions was performed among the infected family members. RESULTS: There was higher HGV viremia in children from the index mothers with HGV infection than those from mothers in control (3/11 vs. 0/14, p < 0.05). Nucleotide sequences analysis showed higher similarity between mothers and their children in the same family (98.6% to 100%) than between mothers of different families (87.8% to 92.4%). The phylogenetic construction also demonstrated distinct sequence clusters for mother and child/children of each family. CONCLUSIONS: The relationship of HGV transmission between mother and children was closely related in family. Spread of HGV might occur intrafamilially. PMID- 12630013 TI - Does end-stage kidney failure influence hepatitis C progression in hemodialysis patients? AB - BACKGROUND/AIMS: The influence of end-stage kidney failure on the progression of liver disease in patients infected with hepatitis C virus and treated with hemodialysis is still controversial. METHODOLOGY: Liver histology of 154 hepatitis C virus infected non-uremic patients was compared with liver histology of 13 hepatitis C virus infected uremic patients treated with hemodialysis. RESULTS: In either group of altogether 167 patients, no normal liver histology was found. Each patient had at least a low-grade lobular and/or portal inflammation. However, statistically significant differences were observed between hepatitis C virus infected uremic and non-uremic patients in the extent of intralobular changes, portal inflammation, and degree of fibrosis. CONCLUSIONS: Non-uremic hepatitis C virus infected patients appear to have more active and progressive liver disease than hepatitis C virus infected patients on hemodialysis. Regular follow-up of uremic patients, associated with earlier detection of hepatitis C virus infection, so as suggested uremia-associated impaired immunoreactivity and increased levels of hepatocyte growth factor described recently, might be implicated in a more favorable course of hepatitis C virus infection in uremic patients. In addition, due to the absence of normal liver histology in either group of hepatitis C virus infected patients, we propose liver biopsy to be mandatory in all these patients, provided that no contraindications exist clinically. PMID- 12630014 TI - The effect of FR167653 in a canine total hepatic vascular exclusion model. AB - BACKGROUND/AIMS: Though liver grafts from non-heart-beating donors are now attracting much attention, these grafts inevitably suffer from severe warm ischemia. The aim of this study was to evaluate the effect of TNF-alpha and IL-1 suppression on warm ischemia-reperfusion injury in a canine total hepatic vascular exclusion model. METHODOLOGY: Warm ischemia was induced by 1-h total hepatic vascular exclusion with active splenofemuro-juglar bypass. Animals were divided into two groups. FR167653 (1 mg/kg/hr) was administered via the portal vein from 30 min prior to ischemia until 2 h after reperfusion to the FR group (n = 7), and a vehicle was administered to the control group (n = 7). The serum alanine aminotransferase, aspartate amino-transferase, lactate dehydrogenase, and hyaluronic acid levels were measured. Hepatic tissue blood flow was also measured. Liver specimens were harvested for histological study, and polymorphonuclear neutrophils were counted. RESULTS: Serum liver enzymes were significantly (p < 0.05) lower, and hepatic tissue blood flow was kept significantly (p < 0.05) better in the FR group than in the control. Histological tissue damage was mild, and polymorphonuclear neutrophil infiltration was significantly (p < 0.05) lower in the FR group than in the control group. CONCLUSIONS: FR167653 provides protective effects on hepatic warm ischemic injury in a canine total hepatic vascular exclusion model. PMID- 12630015 TI - Alanine aminotransferase (ALT) levels in a normal population and interferon therapy in chronic hepatitis C patients with normal ALT. AB - BACKGROUND/AIMS: The aims of this study were to determine the distribution of serum alanine aminotransferase levels in a normal population and to clarify whether interferon treatment is justified in HCV-infected patients with persistently normal alanine aminotransferase levels. METHODOLOGY: The distribution of alanine aminotransferase levels was examined among 949 normal subjects who were negative for hepatitis viruses, denied regular alcohol use. Nineteen patients with chronic hepatitis C and persistently normal alanine aminotransferase levels were treated with alpha interferon (six or ten million units thrice weekly for six months). RESULTS: Peaks of alanine aminotransferase distribution among the normal subjects were seen at 16-20 IU/L and 11-15 IU/L in males and females, respectively. Fourteen of the 19 patients who received interferon treatment had favorable factors of response to interferon (eight with low pretreatment virus load, four with HCV genotype 2 and two with both). A sustained virological response was achieved in eight (57%) of 14, and alanine aminotransferase levels decreased significantly to within the normal range after interferon treatment in six of eight. CONCLUSIONS: Patients with chronic hepatitis C and persistently normal alanine aminotransferase levels should be treated with high doses of interferon if they have favorable factors of response to interferon treatment. PMID- 12630016 TI - TT virus infection in patients on maintenance hemodialysis in Korea. AB - BACKGROUND/AIMS: TT virus is a novel DNA virus that is associated with posttransfusion hepatitis. The prevalence, risk factors, and clinical significance of TT virus infection were evaluated in patients with chronic renal failure who are on hemodialysis. METHODOLOGY: Nested polymerase chain reaction was used to test for TT virus DNA in the serum of patients on hemodialysis as well as in healthy controls. RESULTS: TT virus DNA was detected in 15 (20.0%) of the 75 patients on hemodialysis and 10 (13.2%) of the 76 healthy controls (P > 0.05). In chronic renal failure patients on hemodialysis, the prevalence of TT virus did not differ according to the duration of hemodialysis or the amount of blood transfusion. The prevalence of TT virus was higher in IgG anti-hepatitis B core antibody-positive patients than IgG anti-hepatitis B core antibody-negative patients (27.5% vs. 4.2%, P = 0.03). Two (13.3%) of the 15 TT virus-positive and 6 (10.0%) of the 60 TT virus-negative patients showed elevated alanine aminotransferase levels (P > 0.05). CONCLUSIONS: TT virus infection did not occur more frequently in patients on hemodialysis than in healthy controls. No relationship was found between TT virus infection and liver disease. The correlation between TT virus infection and IgG anti-hepatitis B core antibody suggests that TT virus may share some routes of transmission with hepatitis B virus. PMID- 12630017 TI - Preoperative estimation of risk in hepatectomy using technetium-99m-galactosyl human serum albumin receptor amount by nonlinear 3-compartment model. AB - BACKGROUND/AIMS: Recently, we developed the method for measurement of the technetium-99m-diethylenetriamine-pentaacetic acid-galactosyl human serum albumin (Tc-GSA) receptor amount (R0) using a nonlinear 3-compartment model. We examined the usefulness of R0 for preoperative estimation of risk in hepatectomy. METHODOLOGY: Sixty-three patients who underwent hepatectomy in our hospital were examined for R0. These patients consisted of 26 cases of normal liver, 16 cases of liver fibrosis, and 21 cases of cirrhosis. R0 was measured by the nonlinear 3 compartment model of ligand-receptor binding without blood sampling in Tc-GSA scintigraphy. The expected remnant liver R0 after hepatectomy was calculated from CT volumetry before hepatectomy. RESULTS: The preoperative mean R0 of liver was 15.8 +/- 3.8 mg for Z0, 13.8 +/- 3.9 mg for Z1, and 6.9 +/- 2.3 mg for Z2. R0 in Z2 was significantly lower than Z0 and Z1 (p < 0.0001). Every patient whose remnant liver R0 was over 5 mg tolerated hepatectomy without any postoperative complications. Among 63 cases, 5 patients developed postoperative complications (two liver failures, two postoperative jaundice and one hepatic coma), and remnant liver R0 of these patients were under 5 mg. CONCLUSIONS: From these results, it can be seen that R0 of remnant liver is a useful parameter to decide indication of hepatectomy and predict postoperative complications. PMID- 12630018 TI - Peripheral blood T, B, and NK cells in relation to histological hepatitis activity and fibrosis stage in chronic hepatitis C. AB - BACKGROUND/AIMS: Many data on the pathogenesis of chronic hepatitis C have pointed to host's immune system disorders and a high variety of virus. However, there are no known criteria that could prognose the course of chronic hepatitis C infection. The analysis of T and B lymphocyte subpopulations in the peripheral blood was undertaken in patients with chronic hepatitis C of more than 6 months of duration. METHODOLOGY: Fluorescein isothiocyanate or phycoerythryne conjugated monoclonal antibodies for CD3+, CD4+, CD8+, CD19+, CD3++ HLA DR+, CD16++ CD56+ were used. The correlation between histological hepatitis activity and fibrosis (according Scheuer's scale) and the distribution of lymphocytes in the peripheral blood was sought. RESULTS: All patients with chronic hepatitis showed statistically significant increase in active lymphocytes CD3++ HLA DR+ and CD16++ CD56+ NK cells in peripheral blood. We observed the correlation between these cells and histological hepatitis activity and fibrosis. There was no correlation between the value of CD3+ and CD8+ cells and the stage of liver failure. In the early stage of chronic hepatitis C we noted decrease CD4+ cells with increase B cells CD19+. CD4+/CD8+ ratio was maintained as slightly decreased in chronic hepatitis C in favor of lymphocytes CD8+. CONCLUSIONS: The results show the correlation between peripheral blood value of activated T cell (HLA DR+) and NK cells with histological activity and fibrosis in chronic hepatitis C. Lymphocyte T (CD4+, CD8+) and B (CD19+) did not correlate with grade and stage of hepatitis C. PMID- 12630019 TI - Transcatheter arterial chemoembolization (TACE) in treatment of hepatocellular carcinoma. AB - BACKGROUND/AIMS: Substantial differences about the efficacy of transcatheter arterial chemoembolization for the treatment of hepatocellular carcinoma are reported in literature. This probably depends on the fact that in each single study, different patient selection criteria, type of epidemiological approach, end points adopted and kind of technical approach were used. This study aims to evaluate the efficacy of segmental transcatheter arterial chemoembolization in amelioring patient survival and to determine which patients might really benefit from this treatment. METHODOLOGY: To achieve our goals 193 consecutive patients (110 treated and 83 untreated) were studied. They were selected in the same period of time and matched as far as their demographic and clinical characteristics. RESULTS: Our results demonstrate that both in treated and control patients, Child class, alpha-fetoprotein and tumor diameter significantly influenced survival, resulting important prognostic factors. Transcatheter arterial chemoembolization significantly ameliorated survival in treated patients compared to controls (p < 0.0001). CONCLUSIONS: Transcatheter arterial chemoembolization significantly ameliorates survival in patients with hepatocellular carcinoma. However, the presence of large tumors producing high alpha-fetoprotein levels in patients with advanced Child class should discourage treatment. PMID- 12630020 TI - Hemodynamic and hepatic microcirculational changes in endotoxemic rats treated with different NOS inhibitors. AB - BACKGROUND/AIMS: Severe septic shock may produce hypotension, which is due to the vasodilatational effect of nitric oxide. The effects of different nitric oxide synthase inhibitors on the hemodynamic and hepatic microcirculation of the endotoxemic rats were studied. METHODOLOGY: A prospective controlled study was performed. Eighteen Sprague-Dawley male rats (250-300 g) were anesthetized and studied. The rats were divided into three groups. The rats in group A (n = 6) were injected with lipopolysaccharide (50 mg/kg BW) and L-NAME (5 mg/kg BW). The rats in group B (n = 6) were injected with the same dose of lipopolysaccharide and aminoguanidine (400 mumole/kg BW). The rats in group C rats (n = 6) were injected with same dose of lipopolysaccharide and normal saline as a control. The rats were cannulated with femoral arterial, venous, and jugular venous catheters. Cardiac output was measured using a thermodilutional method, and liver sinusoidal microcirculation was measured with Laser Doppler Flowmetry. The cardiac output, stroke volume, heart rate, blood pressure, and microcirculational flux of the liver in the three groups were measured and compared at 0, 20, 40, 60 and 80 minutes after injection. RESULTS: The rats of group A showed significant decrease of their cardiac output, stroke volume and hepatic microcirculation after the drugs were infused though their blood pressure increased. The rats of group B showed decrease of their blood pressure and stroke volume initially, but no significant change of their cardiac output and hepatic microcirculation. At the 80th min, the rats of group B had the significantly highest cardiac output, stroke volume and hepatic microcirculation among three groups. CONCLUSIONS: The aminoguanidine prevents the hypotensive effect as well as L-NAME during severe sepsis, but it can maintain cardiac output, stroke volume and hepatic microcirculation better than L-NAME. PMID- 12630021 TI - Retrospective monocentric study of 17 patients with adult Still's disease, with special focus on liver abnormalities. AB - BACKGROUND/AIMS: Adult Still's disease is one of the febrile disorders of unknown etiology, characterized by high fever, transient cutaneous rash and leukocytosis. Liver dysfunction in adult Still's disease has been described in some case reports. The objective of this study was to analyze the pattern and the frequency of liver abnormalities in a monocenter series of adult Still's disease patients. METHODOLOGY: Data of 17 patients with adult Still's disease (fulfilling Yamaguchi's diagnostic criteria) were retrospectively reviewed. These patients were followed in an Internal Medicine Department over a period of 7 years. RESULTS: The median age was 27 years with a sex ratio M/F of 1.4. Fever was present in 100% of the cases and hepatomegaly occurred in 47% of the cases. Abnormalities in liver biochemistry, apparent in 76% of the subjects were characterized from moderate (elevation of transaminases between 2 and 5 N) (65%) to severe cytolysis (level of transaminases > 5 N) (12%), cholestasis (elevation of gamma GT and/or PAL) (65%), and increase in the level of LDH (35%). All of these symptoms disappeared either spontaneously or under treatment (83%), within a median period of 18 days. CONCLUSIONS: This study confirms the high frequency of liver dysfunction in adult Still's disease patients. Although it is moderate and asymptomatic in most cases, severe cytolytic hepatitis has been described. This study especially puts forward the need for exploring the possibility of adult Still's disease in the presence of a fever and hepatic cytolysis. PMID- 12630022 TI - Postoperative management for donor safety in living related donor liver transplantation. AB - BACKGROUND/AIMS: Living related donor liver transplantation must hold on in absolute donor safety. Nevertheless, the risks to the donors are increased because of the use of right lobe for size mismatch. Therefore the intensive management for donor operation can hardly be overestimated. In this study, we investigated the utility of prostaglandin E1, methylprednisolone and C-tube in the postoperative management for donor operation based on our understanding of the mechanisms of liver injury after massive hepatectomy. METHODOLOGY: Outcome of the thirteen donors, whose right lobe was removed (right lobe group: 7 cases) or the left lobe only (3 cases) and left lobe plus caudate lobe (3 cases) (left lobe group: 6 cases) from 1999 March to 2000 August, for living related liver transplantation were evaluated. Indication for right lobe living related liver transplantation was basically defined as an estimated left lobe volume/recipient body weight ratio (GVBW ratio) of < 0.8% (range: 0.65-1.4, right lobe group: 1.05 +/- 0.18, left lobe group: 0.89 +/- 0.16). RESULTS: The donors recovered from the operation without the differentiation between right and left lobe group. There were no significant differences in the operative time, the operative hemorrhage, the hospital stays and the postoperative liver function in the two groups. They had no bile leakage and no serious complications. We experienced the left lobe donor who suffered from a cholangitis without any infection and he recovered by the administration of prostaglandin E1 and steroid. They improved bile secretion and bilirubinemia. Eleven recipients (84.6%) were successfully treated. The two causes of death were the sepsis (right lobe) and brain hemorrhage (left lobe). CONCLUSIONS: The current series of right lobe donation was able to show successful results as well as left lobe donation by the postoperative management with steroid, prostaglandin E1 and C-tube drainage based on our understanding of liver injury after massive hepatectomy in the adult-to-adult living donor liver transplantation. PMID- 12630024 TI - Transarterial chemoembolization (TACE) for unresectable hepatocellular carcinoma in cirrhotics: functional hepatic reserve and survival. AB - BACKGROUND/AIMS: Transarterial chemoembolization is widely used for palliative treatment of hepatocellular carcinoma, but patient's characteristics associated with maximal benefit are still undefined. METHODOLOGY: In 81 cirrhotic patients with unresectable hepatocellular carcinoma, who underwent transarterial chemoembolization, variables correlated with survival were studied. In 46/81, the antipyrine metabolism test has been performed before and 72 hours after first transarterial chemoembolization. RESULTS: Mean overall survival of whole population was 22 months. One-, two-, and three-year survival rates were respectively 85%, 38.6%, and 18.1%. Better survival was observed in those patients who received more than one treatment (p = 0.016), while no relationship was found with treatment response, drug used, or number of lobes involved. Univariate analysis of the subgroup with antipyrine pharmacokinetic data revealed a significant relation between survival and baseline albumin (p = 0.039), total cholesterol (p = 0.036), AST (p = 0.017), log of total bilirubin (p = 0.017), and Child-Pugh class (p = 0.029), but not with parameters of antipyrine metabolism. Antipyrine metabolism was not significantly modified by transarterial chemoembolization in the subgroup tested before and after the first treatment. Using Cox regression analysis and selecting AST and log of total bilirubin, a prognostic index was defined: prognostic index 0.006 (AST-83.044) + 0.638 [log of total bilirubin-0.1175]. One-, two-, and three-year survival rates were respectively 92%, 59.2%, and 29.6% for the patient group with prognostic index < 0, and 76%, 14.3%, and 4.8% for the group with prognostic index > 0 (p < 0.01). CONCLUSIONS: Transarterial chemoembolization is a safe procedure and appears beneficial for those patients with a good functional hepatic reserve. The antipyrine metabolism test does not provide any additional prognostic information. PMID- 12630023 TI - Analysis of the effect of IL-12 therapy on immunoregulatory T-cell subsets in patients with chronic hepatitis C infection. AB - BACKGROUND/AIMS: Aim of this study was to investigate the influence of IL-12 therapy on the production of immunoregulatory type 1/type 2-cytokines by peripheral blood mononuclear cells from patients with chronic hepatitis C. METHODOLOGY: Peripheral blood mononuclear cells were isolated from 12 patients with chronic HCV infection before and after eight weeks of IL-12 application (0.03-0.5 micrograms/kg body weight). Peripheral blood mononuclear cells from 40 healthy blood donors served as a control group. The peripheral blood mononuclear cells were incubated for seven days with antigens stimulating specifically type 1 (tuberculin purified protein derivative) or type 2 T-cells (tetanus-toxoid). Furthermore, BCG (bacille Calmette-Guerin) was added to the cultures known to activate macrophages/antigen-presenting cells. Supernatants of peripheral blood mononuclear cells were analyzed for tuberculin purified protein derivative induced production of the type 1 cytokine IFN-gamma, tetanus-toxoid-induced production of the type 2 cytokine IL-5 and the BCG-induced production of TNF alpha by a double sandwich ELISA. RESULTS: IL-12 therapy hardly influenced the regulatory type 1/type 2 T-cell reactivity. In contrast, BCG induced secretion of TNF-alpha was significantly higher after eight weeks of IL-12 therapy (4348 +/- 3083 pg/mL) than before treatment (1559 +/- 988 pgmL, p < 0.01). Clinically, serum alanine transaminase levels significantly decreased during IL-12 treatment but HCV-RNA persisted in all patients. CONCLUSIONS: IL-12 therapy in patients with HCV does not alter the production of regulatory cytokines produced by type 1/type 2 TH cells. The significantly enhanced production of BCG-induced TNF-alpha may, however, indicate an activation of antigen presenting cells or natural killer cells. PMID- 12630025 TI - The changes of hepatic sinusoidal microcirculation and effects of nitric oxide synthase inhibitor during sepsis. AB - BACKGROUND/AIMS: Sepsis may cause changes in liver blood flow, which may result in liver injury. Microcirculation in organ undergoes moderate alteration during sepsis or septic shock. The changes in hepatic microcirculation corresponding to liver functions and the effects of nitric oxide synthase inhibitor on the liver during sepsis were studied. METHODOLOGY: Sepsis was produced by CLP (cecal ligation and two-hole puncture). In part I, the leukocyte adherence, leukocyte rolling numbers, and velocity of sinusoidal microcirculation of liver were compared with in vivo microscopy among early septic, late septic and control rats. In part II, the rats were randomly divided into two groups after CLP procedure, group A was given L-NAME (NG-nitro-L-arginine-methylester hydrochloride), 10 mg/kg BW, and group B was given normal saline as a control. The hepatic microcirculation, measured with a Laser-Doppler Flowmeter, was performed at 0, 2, 4, 6, and 8 hours after the CLP procedure. Their liver functions were also examined and compared with the microcirculation. RESULTS: The results showed that the adherent and rolling numbers of leukocytes in the sinusoidal capillary significantly increased in early and late septic rats; and the centralized velocity of flow significantly decreased in late septic rats. For rats without L-NAME, their sinusoidal flux of the liver increased 2 and 4 hours after CLP, and then decreased gradually. Their GOT levels progressively increased after CLP, but the albumin levels decreased. For rats with L-NAME (group A), their sinusoidal flux levels at 2, 4 and 6 hours after CLP were significantly lower than those in rats without L-NAME (group B), but their GOT levels were higher since the 4th hour after CLP. CONCLUSIONS: Our conclusions are that hepatic microcirculation initially increased then decreased and the liver functions deteriorated gradually after sepsis was induced. These changes were aggravated when the nitric oxide synthesis was inhibited. PMID- 12630026 TI - Impaired antibody response rates after high dose short interval hepatitis B virus vaccination of immunosuppressed individuals. AB - BACKGROUND/AIMS: Because a more rapid and high dose vaccination schedule may be advantageous in immunosuppressed individuals to induce an earlier immunization, the aim of the present study was to assess the response to high dose, short interval hepatitis B virus vaccination in various immunosuppressed-patient populations. METHODOLOGY: A total of one hundred and thirty-eight immunosuppressed patients (86 cirrhotics, 42 dialysis patients, 10 allogeneic hematopoietic cell transplants) and 26 healthy subjects as controls were vaccinated utilizing a high dose vaccine (40 mcg) and a shortened immunization schedule. RESULTS: Ninety-two percent of the controls responded to the high dose, short interval hepatitis B virus vaccination schedule. In contrast, only 48% of the immunosuppressed patients seroconverted to anti-HBs positivity (p < 0.001). No difference in the antibody response rate was seen between the various immunosuppressed populations studied. No significant hepatitis B virus vaccination-related adverse effects were seen in any of the groups vaccinated. CONCLUSIONS: Although a high dose, short interval hepatitis B virus vaccination schedule is safe in immunosuppressed patients, the antibody response is still significantly reduced as compared to healthy subjects and only slightly greater than that achieved with standard vaccination schedules, as reported in literature. The possibility of achieving an earlier immunization, however, may be of some advantage to protect acquired viral infection. PMID- 12630028 TI - Thrombopoietin levels and peripheral platelet counts following living related donor liver transplantation. AB - BACKGROUND/AIMS: Thrombopoietin is the primary hematopoietic growth factor. Thrombopoietin deficiency may cause thrombocytopenia in advanced liver disease. The aim of our study was to investigate the relevance of thrombopoietin levels to peripheral platelet counts in patients with liver disease who underwent LRDLT (living related donor liver transplantation). METHODOLOGY: We divided the six patients who underwent LRDLT into two groups. Group 1 had thrombocytopenia and group 2 had normal platelet counts. We measured serum thrombopoietin and peripheral platelet counts before and after LRDLT. RESULTS: Pre-LRDLT thrombopoietin and peripheral platelet counts were lower in group 1 than in group 2. Thrombopoietin in group 1 significantly increased on the first day after LRDLT and peripheral platelet counts in group 1 increased following the rise in thrombopoietin (p < 0.05). Moreover, a marked increase in thrombopoietin and peripheral platelet counts was found in splenectomized patients during LRDLT. CONCLUSIONS: These findings suggested inadequate thrombopoietin production in advanced stage liver disease which caused thrombocytopenia. Improvement of thrombopoietin production in graft liver function may contribute to increase of peripheral platelet counts. PMID- 12630029 TI - Split liver transplantation for two adult recipients. AB - We report a case of split liver transplantation for two adult recipients. The liver graft (1285 g) was split on the backtable into a right lobe graft (900 g, containing the inferior vena cava and middle hepatic vein) and a left lobe graft (385 g). The right lobe graft was implanted into a patient with hepatitis B cirrhosis uneventfully. The left lobe graft was implanted into a patient with familial amyloid polyneuropathy, but was met with massive bleeding from the transection surface and congestion of segment 4. The dusky appearance of segment 4 disappeared after hepatic artery anastomosis, but Doppler ultrasonography showed reverse blood flow in the segment 4 portal vein. Both patients survived the operation. The case illustrated that a left lobe graft without the middle hepatic vein could be problematic. To benefit two adults with chronic liver diseases, a better design of hepatic vein drainage of segment 4 in the left lobe graft and segment 5 and 8 in the right lobe graft is required. PMID- 12630027 TI - The combination therapy of interferon and amantadine hydrochloride for patients with chronic hepatitis C. AB - BACKGROUND/AIMS: The effect of interferon treatment for chronic hepatitis C patients with genotype 1b virus has been suboptimal. We studied the effect of the combination therapy of interferon and amantadine on patients with a high serum viral load of genotype 1b virus. METHODOLOGY: We studied the virological response of naive chronic hepatitis C patients with a high viral load of genotype 1b virus (4.5 log copies/50 microL or 100 kcopies/mL and higher) during interferon and amantadine administration for 6 months and 6 months after the end of treatment. Twenty patients were treated with interferon alone (natural interferon-beta 6 MU daily for 6 weeks and thrice-a-week for 20 weeks) for 26 weeks. Eleven patients were treated with the combination therapy of interferon and amantadine hydrochloride (100 mg orally daily) for 26 weeks. RESULTS: After daily administration of interferon-beta intravenously once a day for 6 weeks, all patients showed the negative tests of serum HCV-RNA by polymerase-chain-reaction methods by the combination therapy, while 13 patients (65.0%) showed the negative tests by interferon alone (p = 0.0257). At the end of treatment, serum HCV-RNA were not detected in 54.5% of patients treated with interferon and amantadine, while it was detected in 50.0% of patients treated with interferon alone. At 6 months follow-up, only one patient (9.1%) could eradicate HCV-RNA in patients with interferon and amantadine, while no patient could with interferon monotherapy (not significantly). CONCLUSIONS: Amantadine hydrochloride has the additive effects to interferon treatment on the virological responses of serum HCV-RNA during a co-administration, although the combination therapy has not shown a significantly promising effect on the eradication of HCV-RNA in the patients with chronic hepatitis C with a high viral load of genotype 1b virus. PMID- 12630030 TI - Metastatic hepatoma originating from the pons hepatis presenting extrahepatic growth--classification of different patterns covering REX's recessus. AB - A metastatic liver carcinoma which developed from the pons hepatis and presented as an extrahepatic mass on ultrasonography and computed tomography is reported. There have been no other reports of hepatoma documenting the extrahepatic growth developed from the pons hepatis. Herein we study the frequency of the pons hepatis and classify the form of REX's recessus. Of 125 cases that underwent upper abdominal operation, 46 cases (38%) had nothing under REX's recessus (Type I), 48 cases (37%) had a fibrous tissue under REX's recessus (Type II), and 31 cases (25%) had a pons hepatis (Type III). In Type III, some cases suggested that the pons hepatis was ready to disconnect from side to side. We conclude that Type II is a modified form of Type III, and Type I is a modified form of Type II. PMID- 12630031 TI - Double primary mucoepidermoid carcinoma and hepatocellular carcinoma of the liver -a case report. AB - Double primary mucoepidermoid-hepatocellular carcinoma of the liver is extremely rare, and only one case has previously been reported in the literature, although there have been about 14 cases of primary mucoepidermoid carcinoma of the liver. Most of the reported hepatic mucoepidermoid carcinoma showed a poor prognosis. We presently report the second case of a double primary mucoepidermoid carcinoma and hepatocellular carcinoma with a brief review of the published literature. A 52 year-old man was admitted because of epigastric pain that lasted for 2 months. A computed tomography of the abdomen revealed a 7-cm, ill-defined mass with irregular marginal enhancement in the left lobe of liver. Another 2-cm nodular tumor was found in segment 8 of the right lobe. The two separate nodules in the patient's liver demonstrated clearly different histologic and immunohistochemical features. The pathological diagnoses were mucoepidermoid carcinoma and hepatocellular carcinoma for the larger and the smaller tumors, respectively. The patient died of liver failure 6 months after a left lobectomy of the liver. PMID- 12630032 TI - Apoptosis in hepatocellular carcinoma. AB - The process of apoptosis plays a crucial role in several physiological as well as pathophysiological processes, and is a fundamental mechanism for the deletion of unwanted, senescent, or damaged cells. Defects in the apoptotic mechanisms or, even more, disturbances of the equilibrium between cellular replication, growth, differentiation and apoptosis favor the growth of preneoplastic and tumoral lesions in the liver. Clinical and molecular medical analyses have yielded a considerable amount of information about liver oncogenesis. Studies on transgenic models of hepatocarcinogenesis have allowed a meticulous examination of the roles of apoptosis, tumor suppressors, DNA repair processes and other events in the clonal expansion and accumulation of genetic lesions. During initiation, there is rapid proliferation and a high amount of apoptosis, whereas in the promotion stage, there is decreased cell death and increased cell proliferation. Accumulating evidence suggests a defective apoptotic process in human hepatocellular carcinoma. Moreover, a range of prognostic factors related to apoptosis might be used in the early assessment of the progress of hepatocarcinogenesis as well as in hepatocellular carcinoma recurrence. Lastly, many potential therapeutic strategies, that induce the apoptotic process in various ways, can be applied to the hepatocellular carcinoma management. PMID- 12630033 TI - Predicting factors for survival of patients with unresectable pancreatic cancer: a management guideline. AB - BACKGROUND/AIMS: When pancreatic cancer cannot be resected, palliative procedures including gastroenteric bypass and biliary bypass may be selected. However, since predicting survival is difficult, indication of these procedures remains unclear. This study was designed to elucidate the prognostic factors of patients with unresectable pancreatic cancer in order to improve their quality of life. METHODOLOGY: We treated 187 consecutive patients with unresectable pancreatic cancer at the Kobe University Hospital. Fifteen prognostic variables for survival were analyzed (sex, age, the degree of pain, diet, presence of jaundice, main site of the tumor, tumor size, major vessel invasion, liver metastasis, peritoneal dissemination, distal metastasis and operative procedures) in surgically treated patients (n = 125). All patients were followed until death. Cox's proportional hazard model and logistic regression models were used to determine the factors influencing the survival of patients with unresectable pancreatic cancer. RESULTS: Cox's proportional hazard model revealed that duodenal invasion (p = 0.001) and liver metastasis (p < 0.0001) significantly influenced the survival of the patients with unresectable pancreatic cancer. In multivariate logistic regression analysis, liver metastasis (p = 0.009) and peritoneal dissemination (p = 0.004) were significant factors on the six-month survival after palliative operation. CONCLUSIONS: Liver metastasis and peritoneal dissemination were negative predictive factors for the six-month survival of patients with unresectable pancreatic cancer. Palliative bypass surgery is recommended for patients expected to survive long-term (more than six-months). PMID- 12630034 TI - Factors influencing hepaticojejunostomy leak following pancreaticoduodenal resection; importance of anastomotic leak test. AB - BACKGROUND/AIMS: Pancreatic fistula is a common complication after pancreaticoduodenostomy. Prevention of a concomitant bile leak from hepaticojejunostomy is important because it could lead to more serious complications including intraperitoneal abscess, subsequent sepsis and massive hemorrhage by activating pancreatic fistula. This study was designed to determine perioperative risk factors of the hepaticojejunostomy leak for the purpose of decreasing this morbidity. METHODOLOGY: Clinical records of 107 consecutive pancreaticoduodenal resections were reviewed. hepaticojejunostomy anastomoses were performed using absorbable sutures in an end-to-side, single-layer and interrupted fashion. A total of 8 presumed perioperative risk factors were analyzed. They included advanced age, low serum albumin, low serum total cholesterol, impaired glucose tolerance and placement of a biliary drainage catheter as preoperative factors, and dilated common hepatic duct and undone anastomotic leak test as intraoperative factors. In addition, transanastomotic stenting techniques including retrograde transhepatic bile drainage, T-tube and transjejunal drainage were compared with respect to hepaticojejunostomy leak rates. RESULTS: Hepaticojejunostomy leak was demonstrated in 9 patients (8%). Anastomotic leak testing only achieved statistical significance (p = 0.04). It is noteworthy that no hepaticojejunostomy leak developed among 28 patients who underwent this test through a retrograde transhepatic bile drainage catheter. In addition, the frequency of bile leaks (14/107) associated with the transanastomotic stenting techniques urged the necessity of appropriate intraperitoneal drain placement. CONCLUSIONS: Careful anastomotic procedures with a subsequent anastomotic leak test most effectively prevent hepaticojejunostomy leak after pancreaticoduodenal resection. PMID- 12630035 TI - Elevated fasting plasma level of islet amyloid polypeptide (IAPP) in chronic alcoholic pancreatitis (CAP). AB - BACKGROUND/AIMS: The aim of the study was to evaluate fasting plasma level of islet amyloid polypeptide in patients with chronic alcoholic pancreatitis and its possible correlation with the extent of pancreatic structural damage as well as endocrine function. METHODOLOGY: Fifty-seven non-obese patients with chronic alcoholic pancreatitis and 20 healthy subjects were studied. In all subjects RIA plasma fasting amylin, insulin and C-peptide estimation were performed. RESULTS: In patients with chronic alcoholic pancreatitis, particularly in those with advanced pancreatic structural damage in imaging techniques, fasting plasma amylin levels were significantly higher (p < 0.01) than in controls. Within chronic alcoholic pancreatitis group the highest plasma islet amyloid polypeptide values of 139.0 +/- 8.7 pg/100 microL were found in patients with glucose impaired tolerance, which was significantly higher (p < 0.01) than in patients with normal endocrine pancreatic function, non-insulin requiring and insulin requiring diabetes where the respective values were: 69.1 +/- 9.2 pg/100 microL, 75.5 +/- 5.2 pg/100 microL and 84.7 +/- 12.5 pg/100 microL. CONCLUSIONS: Those findings suggest that islet amyloid polypeptide may play a role in the pathogenesis of chronic alcoholic pancreatitis. In addition increased plasma islet amyloid polypeptide level may prove useful as an indicator of an early phase of pancreatic endocrine function impairment and severe parenchymal damage in chronic pancreatitis. PMID- 12630036 TI - Clinical significance of combined pancreas and portal vein resection in surgery for pancreatic adenocarcinoma. AB - BACKGROUND/AIMS: Surgical resection remains the only potentially curative treatment for pancreatic adenocarcinoma for which the resectability and prognosis are still poor. The aim of the present study was to evaluate the efficacy of portal vein resection for pancreatic adenocarcinoma. METHODOLOGY: Between August 1983 and December 2000, 69 patients with pancreatic ductal cell carcinoma underwent resection in our department; 22 of the 69 had combined resection of the pancreas and portal vein. When the pancreas could not be separated from the portal vein, the vein was judged to be invaded by cancer and resected. RESULTS: The mortality rate for portal vein resection was 4.5%, which was similar to that in 47 patients with no resection of the portal vein (2.1%). Postoperative histologic analysis showed that 8 (37%) of the patients who underwent portal vein resection did not have cancer invasion to the portal vein, and 3 of them remain disease free to date. The 3-year survival rate of patients undergoing portal vein resection was 21.3%, and that of patients without portal vein resection was 20.0%. CONCLUSIONS: Resection of the portal vein in cases of pancreatic ductal cell carcinoma has no adverse affect on long-term survival for selected patients. PMID- 12630037 TI - Pylorus-preserving pancreatoduodenectomy associated with longitudinal pancreatojejunostomy for treatment of chronic pancreatitis. AB - Patients with chronic pancreatitis presenting with lesions at the head of the pancreas and in the uncinate process associated with duct dilatation at the body and tail of the gland are inadequately treated by partial pancreatic head resection or by standard pancreatic drainage procedures. A new technique for the surgical treatment of this form of chronic pancreatitis, based on the performance of a pylorus-preserving pancreaticoduodenectomy associated with a longitudinal pancreatojejunostomy, is proposed herein. Nine patients with chronic pancreatitis were operated on according to this technique with no postoperative complications or mortality. Complete symptom control was achieved in all patients. Follow-up periods varied from one month to five years. PMID- 12630038 TI - Endoscopically placed pancreatic stent in a patient with concomitant two locations of main pancreatic duct disruption following pancreatic trauma. AB - Severe pancreatic trauma involving disruption of the main pancreatic duct usually requires operative management. Recently, the successful non-operative treatment of proximal main pancreatic duct injuries by pancreatic stent has been described as having good results. Computed tomography of a 36-year-old man who sustained a blunt abdominal trauma in a motor vehicle accident disclosed a large amount of fluid in the cavity of the lesser sac, as well as a laceration of the pancreatic parenchyma at the pancreatic neck. Without definite evidence of pancreatic duct injury during laparotomy, no pancreatic resection was performed. However, postoperative endoscopic retrograde cholangiopancreatography showed 2 sites of main pancreatic duct disruption at the pancreatic neck and body, respectively. The first attempt for endoscopic pancreatic drainage failed with the endoscopic nasopancreatic drainage tube penetrating into the peritoneal cavity through the distal disruption site. A 5-French pancreatic stent was used for another attempt and successfully passed through the proximal disruption site with its tip located just proximal to distal disruption site. The patient improved steadily and was discharged uneventfully. The presenting case demonstrated that the endoscopically placed pancreatic stent was highly effective for the management of severe pancreatic trauma even though there were 2 sites of main pancreatic duct disruption, and pancreatic resection could be avoided. PMID- 12630039 TI - Pylorus-preserving pancreaticoduodenectomy without homologous blood transfusion in a Jehovah's witness with pancreatic cancer: report of a case. AB - A case of successfully treated pancreatic cancer without homologous blood transfusion in a Jehovah's witness is reported. The patient was a 60-year-old Japanese man and he was diagnosed with pancreatic cancer. Based on the patient's informed consent, human recombinant erythropoietin (12000 IU/day), saccharated ferric oxide (60 mg/day), and 25% human serum albumin (100 mL/day) were administrated for 7 consecutive days prior to the operation. As a result, the hemoglobin level increased by 1.6 g/dL during the week, pylorus-preserving pancreaticoduodenectomy was performed on June 5, 1998. We used a technique of transfusing diluted autologous blood, hypervolemic hemodilution, and a cell-saver device for autologous transfusion during the operation. The operation took 10 hours and 35 minutes, blood loss was 600 mL, and the hemoglobin level at the end of surgery was 12.0 g/dL. He had an uneventful postoperative course, and was discharged on the 31st postoperative day. Although there have been numerous reports on surgery without homologous blood transfusion in Jehovah's witnesses, many of the cases of major surgery have involved cardiovascular procedures. However, major digestive surgery may be feasible in more Jehovah's witnesses if adequate alternative treatments and strict nutritional management are carefully implemented, as in our case. PMID- 12630040 TI - Cutaneous metastasis from pancreatic adenocarcinoma: a case report and a review of the literature. AB - We treated a 77-year-old Japanese man with cutaneous metastases from pancreatic adenocarcinoma. Physical examinations revealed reddish, non-tender and firm nodules in left axilla. Laboratory examination showed a remarkable elevation in the tumor markers of CA 19-9, DUPAN-2 and SPAN-1. Abdominal computed tomography showed a low-density mass in the tail of the pancreas, and histological examination of nodule in the axilla revealed a metastatic adenocarcinoma. The immunohistochemistry of CA 19-9 in the excised nodule was positive. Cutaneous metastasis from pancreatic adenocarcinoma is usually rare, and the prognosis is poor, as the disease is usually advanced. PMID- 12630041 TI - Effect of fluoxetine on symptoms and gastric dysrhythmia in patients with functional dyspepsia. AB - BACKGROUND/AIMS: Although antidepressants have been used for decades in treating patients with functional abdominal syndromes, how they influence gastrointestinal motility remains unclear. We aimed to assess the role of depression in functional dyspepsia, and the effect of antidepressants on the functional dyspepsia patients' symptoms and gastric myoelectrical activity. METHODOLOGY: We conducted an open clinical trial with 40 functional dyspepsia patients. Zung self-rating depression scale was used in evaluating the patients' depression. Cutaneous electrogastrography and evaluation of upper gastrointestinal symptoms were performed before and after administration of a one-month course of fluoxetine. RESULTS: In the baseline study, the depressed functional dyspepsia patients had higher symptom scores than non-depressed patients (P < 0.05). The depressed functional dyspepsia patients had higher percentages of tachygastria than healthy controls (P < 0.05), but the electrogastrography parameters of depressed and non depressed functional dyspepsia patients were not different. After one-month fluoxetine treatment, the symptom scores improved significantly in the depressed functional dyspepsia patients (P < 0.05), but not in the non-depressed patients. Electrogastrography did not improve in either group. CONCLUSIONS: Depressive functional dyspepsia patients had higher symptom scores and responded well to fluoxetine treatment. However, electrogastrography did not improve after the treatment. These findings suggest that depression is significant in the presentation of functional dyspepsia symptoms, but not correlated with gastric myoelectrical activity. PMID- 12630042 TI - Does body mass index (BMI) influence morbidity and long-term survival in gastric cancer patients after gastrectomy? AB - BACKGROUND/AIMS: The long-term survival of patients with gastric cancer is governed by various factors, such as the clinical stage of the cancer, the patient's nutritional state, and the treatment and may be governed by the volume of intraperitoneal adipose tissue. The aim of this study is to clarify the relationship between the degree of the patients' body mass index and their long term survival. METHODOLOGY: Gastric cancer patients who had undergone a gastrectomy with D2-lymphadenectomy and with resection A and B according to the criteria of the Japanese Research Society for Gastric Cancer Rules were subgrouped into those patients with a body mass index < 0.185 (the lower body mass index group) and those patients with a body mass index > 0.210 (the higher body mass index group). The patient's morbidity and long-term survival rate was retrospectively compared between the 2 groups. RESULTS: A significantly longer mean survival rate was observed for the lower body mass index group in stage 2 (1667 vs. 1322 days, P = 0.0240). Also, a significantly longer mean survival rate was observed for the higher BMI group in stage 3a (1431 vs. 943, P = 0.0071). CONCLUSIONS: The body mass index is one of the prognostic factors of stage 2 and stage 3a gastric cancer. However, it does not appear to be useful for determining the prognosis of stage 1a, 1b, 3b, and 4a gastric cancers. PMID- 12630043 TI - Prognostic value of preoperative immunosuppressive acidic protein levels in patients with gastric carcinoma. AB - BACKGROUND/AIMS: Immunosuppressive acidic protein has been found to suppress both phytohemagglutinin-induced lymphocyte blast formation and mixed lymphocyte reaction in vitro. The level of immunosuppressive acidic protein has revealed clear increases with the progression of cancer, and has been reported to be a useful diagnostic and follow-up marker for ovarian and renal cancer. We examined immunosuppressive acidic protein levels in patients with gastric cancer to examine the influence of clinical parameters. METHODOLOGY: The present study was based on a retrospective analyses of data on 107 patients with gastric cancer, all of whom had been surgically treated in the Department of Surgery II, Kyushu University Hospital from 1988 to 1992. All patients were examined for immunosuppressive acidic protein level, based on a peripheral blood sampling done preoperatively. RESULTS: Significant associations among immunosuppressive acidic protein level and clinicopathological parameters, including lymphatic involvement, vascular involvement, and lymph node metastases were evident. The 5 year survival rates were 75.9% and 92.1% for those with immunosuppressive acidic protein level > or = 355 and < 355, respectively, the value being statistically significant (P < 0.05). CONCLUSIONS: These data show that immunosuppressive acidic protein levels may be related to lymph node metastases. Measurement of preoperative immunosuppressive acidic protein levels may be pertinent for the prognosis of patients with gastric cancer and for follow-up clinical management. PMID- 12630044 TI - Effect of anti-Helicobacter pylori IgG antibody titer following eradication of Helicobacter pylori infection. AB - BACKGROUND/AIMS: We measured the levels of IgG antibody in patients treated for H. pylori infection using four commercially available ELISA (enzyme linked immunosorbent assay) kits. METHODOLOGY: A total of 37 patients with peptic ulcers (21 patients had gastric ulcers and 16 patients had duodenal ulcers) were treated and monitored. These patients were treated for 14 days with either a dual therapy regime consisting of amoxicillin at 1,500 mg per day and omeprazole at 20 mg per day or a triple therapy regime consisting of the same medications as above plus metronidazole at 1,000 mg per day. Of these 37 patients, 24 patients (64.9%) had successful eradication of the H. pylori infection. The other 13 patients (35.5%) remained infected. Infection by H. pylori was confirmed prior to treatment using conventional microbiological (rapid urease test) and histological tests on biopsy specimens obtained by endoscopy. Four commercially available ELIZA kits (GAP-IgG, Hel-P, Helico-G and HM-CAP) were used to measure the serum levels of IgG antibody against H. pylori before treatment and at 3, 6 and 9 months after treatment. A significant decrease in the IgG titer was observed with the successful eradication of the H. pylori infection. RESULTS: In patients whom the H. pylori infection was not eradicated, the IgG titer remained the same throughout the testing period. All the kits showed a significant decrease in the IgG levels (approximately 50% at 3 months) with patients that were successfully treated. The HM-CAP kit had the lowest sensitivity at detection the decrease in antibody levels (approximately 42% decrease at 3 months). The GAP-IgG kit was able to detect whether or not H. pylori was eradicated in shortest time period (100% uniformity ratio at 3 months). CONCLUSIONS: The results of this study suggest that monitoring serum IgG levels after treatment may provide an early indicator of the efficacy of therapy in eradicating H. pylori infection. Additionally, the serum IgG level can provide evidence of infection in chronic gastritis patients even when the biopsy specimens are negative by microbiological and/or histological tests. PMID- 12630047 TI - Lofty ambitions. PMID- 12630046 TI - Prognostic value of lymph node staging in gastric cancer. AB - BACKGROUND/AIMS: The latest TNM classification (5th edition) changed the definition of nodal staging from the anatomical localization to the total number of metastatic lymph nodes. This study was designed to evaluate and compare the prognostic significance of nodal staging between the two widely known staging systems, the TNM classification (TNM) and Japanese Classification for Gastric Cancer (JCGC). METHODOLOGY: A total of 582 patients who underwent curative gastrectomy with extended lymphadenectomy for gastric cancer were reviewed retrospectively from hospital records. Based on the localization of metastatic nodes according to the JCGC and the total positive node number according to TNM, the patients were divided into subgroups and their prognoses compared. RESULTS: Lymph node metastasis was found in 189 of the 582 patients (32.5%). Both nodal staging systems were found to be significant prognostic factors by multivariate analysis. A prognostic analysis of the patients by subdivision with the two staging systems indicated that the nodal staging system in TNM was more homogenous than that of the JCGC. CONCLUSIONS: The nodal staging system of the TNM classification is superior to that of the Japanese Classification of Gastric Cancer, because it is simple, reproducible and homogeneous. PMID- 12630048 TI - Mix-sex wards. A gender for change. PMID- 12630045 TI - IGF-1 and IGF-1BP3 in gastric adenocarcinoma. Preliminary study. AB - BACKGROUND/AIMS: Insulin-like growth factor-1 (IGF-1) is a mitogenic and anti apoptotic factor, mainly produced by the liver, which regulates cell proliferation. Most serum IGF-1s are bound with IGF-1BP3. Plasma IGF-1 values are positively related to cancer risk (breast, colon, and lung cancer) and seem to have a prognostic significance in prostatic cancer. The aim of this study is to investigate the relationship between IGF-1, IGF-1BP3 and gastric cancer. METHODOLOGY: IGF-1 and IGF-1BP3 serum levels were measured in 26 consecutive patients (M/F = 15/11, mean age 65 yrs) with histologically proven gastric adenocarcinoma from January 1999 to December 2000. Blood samples were collected at baseline, before surgery with radical intent (total and subtotal gastrectomies + D2 lymphadenectomy), and then at 14th and 50th postoperative days. These values were compared to a control group of healthy people. RESULTS: At baseline was observed a significant increase of IGF-1 serum levels in cancer patients versus control group (p < 0.001). All gastric cancer patients showed IGF-1 over normal limits. After surgery there was a significant decrease of IGF-1 levels (14th day vs. baseline, p = 0.001) that was still present in late postoperative period (50th day). At baseline IGF-1 values were not related to tumor extension or nodal involvement status. Otherwise in postoperative period IGF-1 significantly decreased in earlier stages (N0; T < or = 2) but not in more advanced ones (N+; T > 2). At baseline, IGF-1BP3 values were increased compared to control group but did not significantly decrease after surgery. CONCLUSIONS: IGF-1 values in gastric cancer patients are increased compared to control group, without stratification for stage and nodal status. Moreover radical surgery, with complete tumor ablation, induces a significant decrease in IGF-1 levels, without reach normal limits. Besides at baseline abnormally higher IGF-1BP3 values were observed, suggesting an alteration in IGF-1 and IGF-1BP3 system. PMID- 12630049 TI - Older people. Courtesy entitles. AB - A study of 72 people, with an average age of 72, showed that dignity--and lack of it--were key issues in their estimation of care. Concerns about lack of dignity centred on lack of privacy, mixed sex wards, forms of address and loss of independence. The study suggested that older people do not complain about care for fear of retaliation. PMID- 12630050 TI - Joint-working. Money matters. PMID- 12630051 TI - Training & development. Train of thought. PMID- 12630052 TI - Training & development. Bottoms up. PMID- 12630053 TI - Training & development. Course for celebration. PMID- 12630054 TI - Training & development. The uni cycle. PMID- 12630055 TI - Training & development. Centre of attention. PMID- 12630056 TI - Training & development. Ready, study, go. PMID- 12630057 TI - Training initiative. Power to act. PMID- 12630058 TI - Training & development. Mind games. PMID- 12630059 TI - Small mercies. PMID- 12630060 TI - Sexual healing. PMID- 12630061 TI - Patient access. To the ENT degree. AB - Evaluation of six pilot schemes using specialist GPs to provide ENT services in the community suggests these innovations are popular with patients and have the potential to reduce waiting times. Waiting times in one area, where a specialist GP had been in post for three years, were down to 24 days. The schemes need champions in primary and secondary care and must fit well with the strategy of the local health economy. PMID- 12630062 TI - General practice. All in a day's work. AB - Establishing a first-contact team of two practice nurses and a GP has enabled a general practice to offer same-day access for any patient who wants it. The team provides morning and afternoon sessions five days a week, seeing an average of 45 patients per session. Most conditions seen by the team are self-limiting. The service has freed up GPs to spend more time with patients who have booked an appointment, allowing them to give each patient a minimum of 10 minutes. The first-contact team service has been welcomed by patients. PMID- 12630063 TI - Primary care. Those who can. PMID- 12630064 TI - Open space. Spell it out. PMID- 12630065 TI - Medicines. Back to the future. PMID- 12630066 TI - Data briefing. Target setting. PMID- 12630067 TI - Measurement of steroid levels in saliva in a population-based survey of lifestyle, medical conditions, marriage, sex life and hormone status in aging men: a feasibility study. AB - Some population-based studies on male aging measure testosterone and cortisol in saliva instead of serum, but very few measure estradiol and dehydroepiandrosterone sulfate (DHEA-S), suggesting further testing is needed for reliability and comparative validity. In addition, the effects of interview stress and circadian hormone secretion need to be checked. In a pilot study on the overall sexual capacity of aging men, 48 randomly selected, healthy, heterosexual, cohabiting men aged 50-80 years, from Mannheim, Germany, and 50 from the State College, Pennsylvania, USA, were administered a standardized interview covering medical biography, present and previous life and work, marriage and emotional status. Two saliva samples were collected from each subject for measurement of testosterone, cortisol, estradiol and DHEA-S levels before and after the interview, and each subject completed a confidential self administered questionnaire on intercourse, masturbation, orgasm, fantasies, libido and arousal. Questionnaires, hormone measurement techniques and the survey protocol had been extensively pretested. Prior to the pilot study, the kits for measuring testosterone and DHEA-S in saliva were checked for comparative validity against established measuring techniques in serum in 31 cases for testosterone and in 24 different cases for DHEA-S. These 55 cases underwent clinical diagnosis and were not otherwise involved in this study. The cases had been referred to the Andrology Unit of the University Hospital, Marburg, for reasons unrelated to this study. Given the biological differences for both steroids between their presence in blood and in saliva, a perfect correspondence between the two values was not expected and was not observed. The correlations obtained, however, support the assumption that all statistical relationships between testosterone and DHEA-S values in serum and clinical, as well as behavioral, variables reported to date may be replicated for testosterone and DHEA-S values in saliva. PMID- 12630068 TI - Testosterone suppression in men with prostate cancer is associated with increased arterial stiffness. AB - Low androgen levels in men are associated with increased cardiovascular risk, through unclear mechanisms. We measured arterial stiffness ('compliance') in 21 men receiving complete testosterone suppression therapy for prostate cancer, and in 25 controls. Systemic arterial compliance (SAC), which assesses proximal aortic stiffness, was calculated by simultaneous recording of aortic flow and carotid artery pressure (the 'area method'). Aorto-femoral (A-F), aorto-radial (A R) and femoral-dorsalis pedis (F-DP) pulse-wave velocities (PWVs) were recorded using the 'Complior' system. SAC was significantly lower in the androgen-depleted men compared to controls (0.81 +/- 0.53 vs. 1.18 +/- 0.43 arbitrary compliance units, p = 0.01, mean +/- SD). Correspondingly, their A-F PWV was higher (14.1 (10.1-21.8) vs. 12.4 (9.6-17.4) m/s, p = 0.03, median (range)). Cases tended to be older (75 +/- 7 vs. 71 +/- 6 years, p = 0.07), and to have higher systolic blood pressure (148 +/- 22 vs. 143 +/- 17 mmHg, p = 0.40); however, SAC was still significantly lower (p = 0.03) after adjustment for age and stratification for central systolic pressure (< or = or > the median). Adjustment of A-F PWV for age and central systolic pressure reduced significance to p = 0.07. There was no significant difference in peripheral PWVs between groups. In conclusion, testosterone suppression is associated with increased aortic stiffness, only partly explained by age and blood pressure. Loss of androgens in men might therefore adversely affect cardiovascular risk. PMID- 12630069 TI - The pleiotropic expression of the myotonic dystrophy protein kinase gene illustrates the complex relationships between genetic, biological and clinical covariates of male aging. AB - Aging is a complex process modulated by multiple interactions between environmental and genetic factors. Myotonic dystrophy (DM1) is an autosomal dominant disorder caused by an unstable (CTG)n repeat expansion in the DM1 protein kinase (DMPK) gene. The affected male patients' life expectancy at birth (53.2 years) is more than two decades below that observed in most occidental populations. The DMPK gene expression is pleiotropic and includes the premature expression of several age-related signs, symptoms and metabolic disturbances including hormonal dysfunctions, progressive decrease in muscular mass, presenile cataracts, alopecia, reduced alertness, insulin resistance, dyslipidemia, erectile dysfunction and hypogonadism. The aim of this study was to analyze the relationship between aging covariates and the severity of DM1 expression in 136 DM1 male subjects. DM1 clinical expression was assessed on a validated neuromuscular disability rating scale and was correlated with plasma total testosterone (rs = -0.31, p < 0.001), luteinizing hormone (LH) (rs = 0.52, p < 0.001) and follicle stimulating hormone (FSH) (rs = 0.54, p < 0.001) levels. Following LH releasing hormone stimulation, FSH and LH concentrations increased as a function of DM1 severity (p < 0.05). Muscular disability in DM1 was also positively associated with fasting plasma insulin and triglyceride concentrations (p < 0.05). The association of plasma apolipoprotein B and low-density lipoprotein cholesterol levels with DM1 was not linear across their distribution and tended to reflect cell membrane damage progression. These results suggest that DM1, a simple Mendelian trait, can represent a valuable model to illustrate the complex relationships between variables associated with male aging. PMID- 12630070 TI - Association of DHEA-S and estradiol serum levels to symptoms of aging men. AB - OBJECTIVES: A number of interactions between age-related changes in serum levels of dehydroepiandrostendione sulfate (DHEA-S) and estradiol and symptoms of aging men have been proposed, yet data regarding this issue are scant. We therefore set up a prospective study to analyze these associations. METHODS: In a prospective, cross-sectional study, men aged 45-85 years were recruited. All men completed a questionnaire containing 38 items covering a number of aspects of the aging male. Questionnaires were compiled by using items from previously published and validated questionnaires. Several socioeconomic parameters were also determined. In parallel, serum levels of testosterone, free testosterone, luteinizing hormone (LH), follicle stimulating hormone (FSH), DHEA-S, estradiol, sex hormone binding globulin and prostate-specific antigen (PSA) were quantified by commercially available immunoassays. RESULTS: A total of 375 men with a mean age of 59.9 +/- 9.2 years (mean +/- standard deviation) were analyzed. Average DHEA-S and estradiol levels of 135.8 +/- 90.9 micrograms/dl and 29.7 +/- 14.6 pg/ml, respectively, were recorded. DHEA-S serum levels were negatively correlated to patient age, sexual function score, total score and PSA. Estradiol serum levels were positively correlated to testosterone and free testosterone. None of the other scores or questions revealed a correlation with DHEA-S or estradiol serum levels. CONCLUSION: This prospective study elucidates only small interactions between partial androgen deficiency of the aging male (PADAM)-related symptoms and serum levels of DHEA-S and estradiol. Nevertheless, the data suggest an impact of DHEA-S on sexual function. PMID- 12630071 TI - Erectile dysfunction and its neuroendocrine correlates: towards an integral perspective. PMID- 12630072 TI - The effects of testosterone deficiency on male sexual function. PMID- 12630073 TI - Is there a role for estrogens in the maintenance of men's health? AB - This paper gives an overview of our own studies and the literature on the biosynthesis and metabolism of estrogens in elderly men, the estrogen action in the male, and the clinical usefulness of estrogen therapy, including the phytoestrogens. Finally, the paper includes a short review of our knowledge of xenoestrogens and men's sexual health. A strong estrogen-deficient status is seen in male patients with mutations of the estrogen receptors or in cases of deviations of the aromatase gene. On the other hand, there are no clear age dependent changes in estrogen secretion. But, in men with disorders of glucose metabolism and also of increased body mass index, the serum estrogen concentrations are significantly elevated. There are also strong positive correlations between serum estrogen levels and bone density, including prevalence of fractures and mood in men. New fields of interest are natural fatty esters of endogenous estrogens, e.g. lipoprotein-associated estrogens, and the role and clinical significance of tissue-specific, local estrogen biosynthesis (e.g. different promoters of the aromatase gene). Exogenous estrogen treatment is focused today on patients with normal testosterone and low levels of circulating estrogens documented on several occasions and with clinical symptoms of hormone deficiency; male-to-female transsexuals; and selected patients with prostate cancer. Some clinical studies show the benefits of estrogen treatment on some cardiovascular parameters and for treating selected signs of mental stress. An indirect estrogen replacement can occur if dehydroepiandrosterone is given orally to men. The clinical usefulness of dissociated estrogens, including non feminizing estrogens and selective estrogen receptor modulators, is still an open question. The beneficial action of phytoestrogens in lowering the clinical symptoms of benign prostatic hyperplasia is well documented. Finally, the question about the definitive influence of so-called endocrine disruptors (xenoestrogens) on sexual functions in men is also discussed. PMID- 12630074 TI - Growth hormone, somatomedins and men's health. AB - According to current basic science as well as the clinical literature, human growth hormone has become an important topic in the field of anti-aging medicine. It is well known that the administration of human growth hormone compensates for the dwarfism syndrome in growth hormone-deficient children and growth hormone is also established as a substitution therapy for adults with pituitary deficiency. The effects mediated by growth hormone comprise an increase in muscle mass, a decrease in body fat, improved physical condition, oxygen consumption and overall quality of life, an improvement of the ratio of high-density lipoprotein to low density lipoprotein, as well as an increase in bone density. Due to the insulin antagonistic effect of growth hormone as well as its insulin-like growth factor-1 mediated mitogenic effect, there is reason to doubt the safety of administering growth hormone outside the framework of approved indications. In an ongoing study, we are evaluating whether growth hormone and somatomedins might be of importance in the regulation of male sexual performance, including penile erection. PMID- 12630075 TI - Digital images for interobserver variability comparison in cervicovaginal cytology. AB - OBJECTIVE: To assess whether digital images can be useful in evaluating interobserver variability in cervical-vaginal cytology. STUDY DESIGN: In phase 1 of the study, to measure interobserver variability, a set of 160 digital images was submitted to 192 cytologists with experience ranging from 2 to > 30 years. The set was preceded by two days of immersion in lessons and practical exercises with digital images. In phase 2, to compare different procedures of interobserver variability, two different sets of slides and one set of digital images were used. RESULTS: In phase 1, kappa and weighted kappa w values computed against both the consensus and the target diagnosis showed good agreement, with few exceptions. In phase 2, the consensus and target diagnoses obtained on the slide sets and digital set were compared. Mean k and kw values obtained with the digital images in phase 2 were significantly lower as compared to those in phase 1. CONCLUSION: A set of digital images can be a useful tool for evaluating and improving interobserver reproducibility. A two-day course on digital images could be an ideal modality for introducing this new technology. PMID- 12630076 TI - P16INK4A as an adjunct test in liquid-based cytology. AB - OBJECTIVE: To assess the utility of P16INK4A as an adjunct test in liquid-based cytology in cases with equivocal morphologic changes of high grade squamous intraepithelial lesion (HSIL). STUDY DESIGN: P16INK4A immunoreactivity was investigated in residual ThinPrep material (Cytyc Corp., Boxborough, Massachusetts, U.S.A.) from 30 cases with equivocal diagnoses of HSIL that had corresponding follow-up biopsies. Two control ThinPrep cases were included: 1 HSIL with biopsy-confirmed cervical intraepithelial neoplasia (CIN) 3 and a negative specimen with a corresponding biopsy of squamous metaplasia. The expression of P16INK4A in ThinPrep specimens and corresponding biopsies was scored as previously described. A ThinPrep case was scored positive if it contained > 10 abnormal cells with nuclear and cytoplasmic immunocytochemical staining. Corresponding biopsies were scored as having negative, sporadic, focal or diffuse staining. RESULTS: The P16INK4A antibody assay was positive in 19 of 30 ThinPrep cases (63.3%). Seventeen of the 19 (89.4%) biopsies corresponding to the positively stained ThinPreps also were positive, with a score of at least focal positivity in the dysplastic regions (2 CIN 1, 4 CIN 2, 11 CIN 3; 2 lesions lost in the tissue recut). The assay was negative in 11 ThinPreps (36.6%) and 10 biopsies (33.3%) with tissue confirmation of chronic cervicitis (5), squamous metaplasia (2), CIN 1 (3) and 1 lesion lost in the tissue recut. Seventeen of 18 (94.4%) ThinPreps confirmed as high grade lesions upon biopsy showed P16INK4A positivity. The control HSIL case with a CIN 3 biopsy was diffusely positive for P16INK4A, and the control negative case with biopsy diagnosis of squamous metaplasia was negative. Nondysplastic squamous and metaplastic epithelium in 7 biopsies and nondysplastic squamous or metaplastic cells in ThinPrep cases were negative. Sporadic staining of bacteria, inflammatory cells and endocervical cells was noted. CONCLUSION: ThinPrep cases in the equivocal cytologic category with the corresponding tissue biopsy assayed for P16INK4A expression showed that there was utility for this type of testing. A larger series comparing corresponding ThinPrep and tissue biopsies will be undertaken. The role of HPV infection in these cases will also be explored. PMID- 12630077 TI - Topometric analysis of diffuse astrocytomas. AB - OBJECTIVE: To investigate differences between astrocytomas of WHO grade 2 and anaplastic astrocytomas of WHO grade 3 in terms of topometric variables characterizing individual tumor cell nuclei. STUDY DESIGN: Paraffin sections from surgical specimens from 25 astrocytomas (grade 2, n = 11; grade 3, n = 14) were analyzed by means of an image analysis system. At least 300 tumor cell nuclei were measured in the region with the highest Ki-67 proliferation index. Three different kinds of topometric variables were determined for each tumor cell nucleus: (1) several distances; (2) the variable Angle 2/1, the angle between the straight lines representing the distance to the nearest and second-nearest nucleus; and (3) the number of neighbors according to our mathematical definition. RESULTS: Most topometric variables showed distinct differences between the 2 tumor grades (multivariate analysis of variance), with 88% cases correctly reclassified by means of cross-validated discriminant analysis. The variables with the highest discriminatory power were the SD of Angle 2/1 and the ratio between the distance to the second-nearest and nearest tumor cell nucleus, with lower values for these variables in anaplastic astrocytomas. Even variables concerning neighborhood relationships showed significant differences. CONCLUSION: The results of this pilot study show that this first set of topometric variables is sufficient to detect differences between topologic characteristics of tumor cell nuclei in astrocytomas grade 2 and grade 3. Topometry seems to be an important tool for grading astrocytomas. PMID- 12630078 TI - Computer-assisted analysis of p53 and PCNA expression in oral lesions infected with human papillomavirus. AB - OBJECTIVE: To carry out a retrospective study to determine whether human papillomavirus (HPV) infection and immunohistochemical expression of p53 and proliferating cell nuclear antigen (PCNA) are related to the risk of oral cancer. STUDY DESIGN: Fifty-seven oral biopsies, consisting of 30 oral squamous papillomas (OSPs) and 27 oral squamous cell carcinomas (OSCCs) were tested for the presence of HPV 6/11 and 16/18 by in situ hybridization using catalyzed signal amplification and in situ hybridization. p53 And PCNA expression was analyzed by immunohistochemistry and evaluated quantitatively by image analysis. RESULTS: Nineteen of the 57 oral lesions (33.3%) were positive for HPV. HPV 6/11 was found in 6 of 30 (20%) OSPs and 1 of 27 (3.7%) OSCCs. HPV 16/18 was found in 10 of 27 (37%) OSCCs and 2 of 30 (6.7%) OSPs. Sixteen of the 19 HPV-positive cases (84.2%) were p53 negative; 5 (9%) were HPV 6/11 and 11 (19%) HPV 16/18, with an inverse correlation between the presence of HPV DNA and p53 expression (P = .017, P < .05). PCNA expression appeared in 18 (94.7%) of HPV positive cases, showing that HPV 16/18 was associated with intensity of PCNA expression and with OSCCs (P = .037, P < .05). CONCLUSION: Quantitative evaluation of p53 by image analysis showed an inverse correlation between p53 expression and HPV presence, suggesting protein degradation. Image analysis also demonstrated that PCNA expression was more intense in HPV DNA 16/18 OSCCs. These findings suggest involvement of high-risk HPV types in oral carcinogenesis. PMID- 12630079 TI - Nuclear chromatin texture in rectal cancer. Relationship to tumor stage. AB - OBJECTIVE: To characterize, by morphometric and chromatin texture analysis, a series of rectal carcinomas classified according to Dukes staging. STUDY DESIGN: High-resolution imagery of 6,001 nuclei from 51 specimens of rectal carcinoma and 22 specimens of normal rectal tissue was digitally recorded. A set of 93 features descriptive of the spatial and statistical distribution of nuclear chromatin was computed for each nucleus to form a characteristic signature. RESULTS: Rectal carcinomas were significantly different from normal rectum in their digital signature. Eleven karyometric features, such as nuclear area and total optical density, were clearly different between the groups, with significant differences found in analysis of 8 of those features. The most distinctive pattern in lesion signatures in comparison with normal rectal tissue was observed at Dukes' stage D. However, the highest average signature values were seen at Dukes' stage B. The lesion signatures and total optical density observed in cancer specimens deviated markedly from values in the normal group. CONCLUSION: Chromatin texture signature proved to be a useful method of identifying and characterizing nuclear differences between rectal carcinoma and normal rectal tissue. PMID- 12630080 TI - DNA ploidy in gastrointestinal B-cell lymphomas. An image analysis study of 43 cases. AB - OBJECTIVE: To analyze the prognostic importance of DNA ploidy pattern on gastrointestinal (GI) B-cell lymphoma using image cytometry (ICM) and to compare the results with previously published flow cytometry (FCM) data. STUDY DESIGN: Forty-three cases of surgically resected primary GI B-cell lymphomas were examined. Thirty-eight tumors were located in the stomach, 2 in the small intestine, 1 in the large bowel and 2 in both the stomach and small intestine. Six cases were at stage E I 1, 15 at stage E I 2, 20 at stage E II 1 and 1 each at stages III and IV. Histologically, the lymphomas were classified as GI low grade marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT) type (low grade, 12 cases), low grade MALT lymphoma with a high grade component (mixed type, 10 cases) and GI diffuse large B-cell lymphoma (DLBC) (high grade MALT lymphoma, 21 cases). After gross removal of nonneoplastic tissue, single cell suspensions were prepared from paraffin blocks and stained according to Feulgen. Ploidy analysis was done using a custom-made DNA cytometer and Optimas image analysis software (Optimas Corp., Seattle, Washington, U.S.A.). RESULTS: Aneuploidy was found in 42% (5/12 cases) of low grade MALT lymphoma, 90% (9/10 cases) of mixed type lymphoma and 100% (21/21 cases) of GI DLBCL. DNA ploidy had no significant impact on overall survival time (P = .73). CONCLUSION: ICM analysis showed a higher proportion of aneuploidy in GI lymphomas as compared to that in prior studies using FCM for ploidy determination. Whether DNA ploidy is an independent prognostic factor remains to be determined. PMID- 12630081 TI - Cytologic appearance of toxic nodular goiter after thyrostatic treatment. A karyometric study. AB - OBJECTIVE: To assess whether the cytologic appearance of aspirates from toxic nodular goiter is substantially modified during the course of therapy with thyrostatic drugs. STUDY DESIGN: Morphometric features of thyrocyte nuclei in aspirates obtained from nontoxic nodular goiter (NTNG), toxic nodular goiter before treatment (TNG-untreated) and toxic nodular goiter during thyrostatic administration (TNG-treated) were examined. The relationship between the degree of morphologic changes and the duration of therapy was evaluated. An analysis of the composition of aspirates was also performed. RESULTS: The sizes of thyrocyte nuclei in the TNG-untreated group were larger than in the NTNG and TNG-treated groups, and treatment with thyrostatics was accompanied by a gradual decrease in the sizes of thyrocyte nuclei. However, karyometric features showed a tendency to increase again in patients treated for longer than 1 year, with the variability of nuclear size in a smear (anisokaryosis) increasing more markedly than the mean size of nuclei. Moreover, in those patients, nuclei with visible nucleoli were found. CONCLUSION: Only long-term therapy with thyrostatic drugs leads to changes in the microscopic appearance of smears obtained by fine needle aspiration biopsy (FNAB) of the thyroid relevant to cytologic diagnosis. Thus, FNAB can be performed successfully after the onset of treatment with thyrostatics if the cytologist is informed of the time scale of treatment in each case. PMID- 12630082 TI - Effect of freezing on histologic grading of invasive ductal breast cancer. AB - OBJECTIVE: To quantify the histologic changes caused by freezing during tissue processing and their influence on histologic malignancy grading as a prognostic factor in invasive ductal breast cancer. STUDY DESIGN: We studied frozen and nonfrozen formalin-fixed, paraffin-embedded samples of 18 cases of invasive ductal breast cancer. Features associated with histologic malignancy grading of breast cancer--i.e., nuclear pleomorphism, mitotic index and tubular differentiation--were assessed by quantitative morphometric methods. RESULTS: In our material, frozen samples consistently had a smaller mean nuclear profile area than nonfrozen samples (mean difference, 32%). Frozen nuclei were also clearly less symmetric and uniform in shape than non-frozen nuclei. Moreover, frozen samples had consistently higher mitotic indices than nonfrozen samples (mean difference, 66%, with the standardized mitotic index). Tubular differentiation, as expressed in fraction of fields with tubular differentiation, increased by 16% as a result of sample freezing. CONCLUSION: According to our results of morphometric measurement in invasive ductal breast cancer, great caution should be exercised when prognostic conclusions are based on frozen tissue samples. PMID- 12630083 TI - Validation of the novel indices of nuclear pleomorphism, polarity and spatial distribution in the grading of urothelial carcinoma. AB - OBJECTIVE: To evaluate the role of the novel histomorphometric variables of pleomorphism, orientation and variability of spatial cytologic organization to objectively quantify histopathologic grade in urothelial carcinoma. STUDY DESIGN: As a validation model, we chose 51 papillary urothelial carcinomas. Thirty-six (70%) were low and 15 (29%) high grade tumors (World Health Organization grading system). Thirty-one cases (61%) were noninvasive, 12 (23%) exhibited lamina propria invasion, and 8 (16%) invaded the deep smooth muscle. Histomorphometric measurements were performed on noninvasive areas only. Pleomorphism was characterized by the anisokaryosis index (AIX), based on the autocorrelation function applied to classic nuclear shape descriptors. Additional indices of pleomorphism included standard deviations (sd) and coefficients of variation of these shape descriptors. Loss of polarity was assessed by the orientation index (ORX), based on Fourier transformation. Intraepithelial nuclear spatial distribution index (NSDX) was also computed. RESULTS: Low grade tumors exhibited less pleomorphism, higher orientation indices and a more homogeneous spatial distribution than did high grade tumors (area-AIX: P = .004, ORX: P < .0001, NSDX: P = .001). Multivariate analysis revealed the significant discriminators of grade to be nuclear size and orientation (size: max-diam, P < .0001; width: P < .0001; orientation: ORX: P < .0001). A discriminant score combining these independent variables distinguished between low and high grades in 98% of cases. The method was successfully validated using a testing sample of 40 new patients (accuracy, 94%). Lamina propria invasion was independently predicted by nuclear pleomorphism (width-sd: P = .0001, sensitivity = 64%, specificity = 91%). Muscle invasion was independently predicted by nuclear area (P < .0001) and pleomorphism (area-sd: P < .0001, max-diam-sd: P < .0001, width-AIX: P < .0001, sensitivity = 87%, specificity = 100%). CONCLUSION: These novel morphometric methods may serve as more objective methods of histopathologic grading and may contribute to the development of automated systems for quantitative grading of stratified and transitional epithelial neoplasms. PMID- 12630084 TI - ["Invisible hand of a reviewer"--peer review of papers in "Morfologia" journal. Current practice and perspectives]. PMID- 12630085 TI - [Ultrastructure of the cerebral cortex after exposure to gas containing hydrogen sulfide]. AB - In experimental animals exposed to hydrogen sulfide-containing gas, electron microscopic study of cerebral cortex indicated the variability in the reaction of layer V neurons: along with neurons demonstrating irreversible damage, some cells with single signs of alteration were found. The reaction of different types of glial cells was also non-uniform. During initial 12 h following single exposure, oligodendrocytes showed the signs of intensified endocytosis. Following repeated exposures, severe destructive changes were found in myelinated nerve fibers and in single neurons, in which the karyolysis and cytoplasmic swelling were observed. PMID- 12630086 TI - [Formation and structural organization of a barrier on the brain outer surface]. AB - Using the methods of light and electron microscopy and immunocytochemistry, the barrier structures on the outer surface of the forming human brain were studied in the early prenatal period of development. The data obtained suggest that the superficial barrier in the forming brain is heteromorphous and consists of differently specialized morpho-functional complexes. The areas of contact of developing brain surface with blood capillaries of pericerebral vascular network were found to be the zone of the blood-brainbarrier formation, while the areas of the surface bordering the meningeal extracellular matrix were the zone of future liquor-brain barrier. PMID- 12630087 TI - [Morpho-functional changes in mauthner neurons of goldfish adapted to prolonged orthodromic stimulation of acoustic nerve in vitro]. AB - In newly-hatched goldfish the adapted state was developed using repeated short term stimulation of the vestibular apparatus. It differed from the non-adapted state by the absence of fatigue during the turns of fishes produced by unilateral tail fin flicks in ring channel. Electrophysiological study of Mauthner neurons (MN) in adapted goldfish using the slices of medulla oblongata incubated in vitro, has shown the preservation of the amplitude of the extracellular responses after prolonged orthodromic electric stimulation of acoustic nerve, which suppressed the responses of non-adapted MN. Thus, the resistance against fatigue has developed also at the level of MN. Using electron microscopy it was demonstrated that adapted MN following the prolonged electric stimulation in vitro were characterized by the resistance of specialized desmosome-like contacts, similar to what was earlier observed in these neurons in vivo. The data obtained suggest that indirect evaluation of MN functional state in goldfish by counting the number of turns performed by it in narrow ring channel, corresponds to the direct morpho-functional assessment of MN state. This form of innate behavior of freely moving fish seems to be the result of MN excitation. PMID- 12630088 TI - [Surgical micro-neuroanatomy of the formations in the posterior cranial fossa as applied to retrosigmoid and transcondyloid approaches]. AB - Modern concepts on the surgery of the tumors of the cranial basis and those of posterior cranial fossa, in particular, reside on the expertise in microarchitecture of the region. The method of injection in brain vascular system using a marked block preparation enabled to preserve anatomically correct correlations of nerves and blood vessels, as well as of other brainstem structures, and to study the microtopography of this region using the microscope and microinstruments. The study was performed using 10 block preparations of posterior cranial fossa. The microneuroanatomy of middle and lower petroclival zones was studied in detail, and the rationality of the usage of suboccipital retrosigmoidal and lateral transcondillary approaches to the pathological structures within this zone is substantiated. PMID- 12630089 TI - [Anatomic and topographical characteristics of the correlations between glossopharyngeal nerve and autonomic nervous system in Canidae and Mustelidae]. AB - Using macro-microanatomical approach, the study of glossopharyngeal nerve was performed on carcasses of fur animals belonging to Canidae and Mustelidae families (mink, sable, sable polar fox, fox). The species peculiarities of interconnections of tympanic nerve with inner carotid nerve and ear branch of vagus nerve were established. The characteristics of the course and connection of the carotid sinus branch of glossopharyngeal nerve with sympathetic branches of cranial cervical ganglion, were defined. The variants of total absence of vagal pharyngeal branch with significant development of similar branch of glossopharyngeal nerve, were noted. PMID- 12630090 TI - [Postmortem and intravital study of the human heart in prenatal ontogenesis]. AB - The objective of this study was to evaluate the growth of the heart in human fetuses at weeks 18-40 of intrauterine development. The investigation included 2 series of observations: 182 hearts were taken from the fetuses at autopsy, while 58 hearts were studied in living fetuses by ultrasonography of the pregnant women. The following parameters were assessed: heart length and width, atrial and ventricular width, diameter of the aorta. The dimensions of the fetal heart were found to grow in proportion to the gestational age, however the growth of individual heart structures was nonuniform. Both intravital and posthumous investigations have established that in the fetuses of this gestational age the width of the right chambers of the heart prevailed over that of the left chambers. PMID- 12630091 TI - [Morphological changes in smooth myocytes of cerebral arteries in experimental hypertension and after its elimination]. AB - Using cytological, histochemical and morphometric methods, the changes of leiomyocytes of cerebral arteries was studied in 55 pups with experimental hypertension induced by coarctation of the aorta and in 15 animals following reversal of coarctation as compared to the normal characteristics of these cells established in 15 control dogs. It was established that hypertension induced hypertrophy and polyploidy of leimoyocytes of the circular muscular layer in the tunica media of the cerebral arteries, that was manifested as the increase in their nuclear DNA content and in the numbers of binuclear cells. After the elimination of hypertension, which was previously established in the cerebral vasculature, the tendency for the reversal of the changes in nuclear and cytoplasmic linear parameters of the cells examined was observed. However, the nuclear DNA content and the number of polyploid leiomyocytes in the cerebral arteries remained practically unchanged. The latter indicates that the polyploidy of leiomyocytes in the cerebral arteries is irreversible. PMID- 12630092 TI - [Structure and cytoarchitecture of the human thymus in adolescents and young adults]. AB - The paper presents the quantitative parameters characterizing the cytoarchitecture of structural and functional components of thymus of practically healthy humans in adolescent and juvenile age periods. It was shown, that despite significant structural reorganization, in adolescence the lymphocytopoietic function of thymus was still high. In juvenile age, structural changes became more intensive. These were accompanied with the disturbances of organ function, manifested by a significant drop in mitotic index and reduction of blast cell content. PMID- 12630093 TI - [Structure of terminal lymphangion of the thoracic duct]. AB - Peculiarities of terminal lymphangion wall in humans and dogs were studied in its different parts using various morphological methods. Main results were obtained with the use of a total preparation method after A.V. Borisov. Heterogenous distribution of myocytes in different regions of lymphangion (muscular cuff, valve sinus wall) was noted. In the terminal lymphangion myocytes from three anatomically and topographically different muscles: cuff muscle, tensor muscle of the distant valve, tensor muscle of the proximal valve (thoracic duct orifice valve). Some features of terminal lymphangion construction similar to those seen in other lymphangions as well as certain specific characteristics and species peculiarities, are described. PMID- 12630094 TI - [Surgical anatomy of superior and mid-thoracic areas of the thoracic duct]. AB - To investigate the peculiarities of surgical anatomy of superior and mid-thoracic portions of the thoracic duct, 70 human corpses of adults of both sexes were studied using modern anatomical and experimental methods. Some regularities of the changes of syntopy of the mediastinal organs and vessels and the connection of these changes with the individual features of thoracic structure are described. The relation between these peculiarities with the quantitative parameters of surgical procedures performed in superior and mid-thoracic portions of the thoracic duct is demonstrated. PMID- 12630095 TI - [Comparative-anatomical study of the formation of small and large intestinal (ileocecal) valve]. AB - The results of the comparative-anatomical investigation of the formation of ileocecal valve are presented. The study was based on the examination of the specimens obtained from 123 representatives of all vertebrate classes and 48 isolated preparations of ileocecal area of the intestine of corpses of humans aged 22-64 years. It was performed using anatomical, histological and morphometrical methods. It was shown that the formation of this closing mechanism followed the route from sphincter between small and large intestine to the development of a shutter and, eventually, of a valve. Using these concepts, the structural variants of ileocecal valve which were detected in humans were analyzed taking into consideration both its extreme variation forms (ileocecal morphological sphincter without the valve and ileocecal valve without the sphincter) and intermediate forms, which should be considered as the variants of norm. PMID- 12630096 TI - [Structural organization of compact and cancellous tissue of human rib in aging]. AB - Using the scanning electron microscopy the thickness of external and internal layers of compact bone of the rib as well as the structure of its cancellous bone was investigated in humans aged from 19 to 89 years. The differences of bone remodeling mechanism were found in men and women of the older age groups. In men, the gradual attenuation of osseous structures was taking place. In women, bone loss was found to be rapidly progressing due to the elimination of its structural elements (transformation of compact layer into cancellous bone and disappearance of some part of trabeculae from the latter). PMID- 12630097 TI - [Architecture of cancellous tissue of the scaphoid bone of the wrist]. AB - The experimental-theoretical studies were performed to investigate the structure of substantia spongiosa of the scaphoid bone of the wrist. Morphological prerequisites for the typical fractures of scaphoid bone and for their delayed consolidation, as well as for the false joint development in some of its zones were found. The peculiarities in the structure of substantia spongiosa associated with the differences of magnitudes and directions of the loads applied to the various regions of scaphoid bone, were distinguished. On the basis of the results obtained, functional zones and two major force vectors were detected, that were shown to determine different biomechanical conditions under which scaphoid bone of the wrist was functioning. PMID- 12630098 TI - [Osteogenesis in healthy children and in children with growth retardation depending on body constitution]. AB - Somatometric and radiographic study of bones of the hand and of the distal forearm was performed in 258 healthy children and adolescents (148 boys and 110 girls) aged 11-17 years and in 115 children (84 boys and 31 girl) aged 13-16 years which were under the observation due to growth retardation. It was shown that within the groups of the same age children with various somatometric characteristics possessed different levels of skeletal (and therefore, biological) maturity. In the group of healthy children, different variants of skeletal maturation were established, which corresponded to the constitutionally stipulated rates of somatic development including accelerated, average and retarded. In children with growth retardation, the third (retarded) variant of somatic development rate was prevalent, which was also found in the group of healthy children and was accompanied with its peculiar osteogenesis characteristics. In the other cases, when such abnormalities of skeletal maturation as asymmetry in the appearance of ossification centers and synostoses as well as the presence of pseudoepiphyses, were observed, growth retardation could be caused by some endocrine pathology. Thus, the radiographic study enables the differentiated approach to the evaluation of the morphological status of children and adolescents and permits to avoid the unjustified administration of hormonal treatment. PMID- 12630099 TI - [Characteristics of body composition of the students of boarding schools]. AB - The article describes the findings on the peculiarities of body composition of pupils of boarding high schools in Vitebsk region in the 1990s. The age- and sex dependent dynamics of body components was shown in children aged 8-15 years in absolute values and as percentage in relation to total the body mass. PMID- 12630100 TI - [Practical recommendations to the preparation of corrosion casts of cerebral vascular bed]. PMID- 12630101 TI - [Indirect aerosol lymphography of intrathoracic lymph nodes in experiment]. PMID- 12630102 TI - [Hypothesis on the role of osteocytes in the formation of a stable morphological structure of bone matrix minerals]. PMID- 12630104 TI - [Concept of "sinew" in the dictionary of Russian Academy]. PMID- 12630103 TI - [Structure and biological role of prostasomes]. PMID- 12630106 TI - [Inauguration of the monument to the outstanding Russian histologist professor Aleksandr Ivanovich Babukhin in his hometown of Orel]. PMID- 12630105 TI - [Sergei Nikolaevich Kasatkin--founder of the department of Human Anatomy in the Volgograd Medical Academy (to the 100 years of birth)]. PMID- 12630107 TI - [Effects of reactive species of oxygen and nitrogen on iron ion release from ferritin and synthesis of dinitrosyl iron complexes]. AB - Using EPR spectroscopy it was established that Fe ions released from ferritin under the action of glutathione and superoxide took part in the formation of dinitrosyl complexes of iron with glutathione (DNIC). The reaction between O2-. and NO resulted in the formation of peroxynitrite, which oxidized glutathione to the thiyl radical. In these conditions, DNIC did not inhibit the formation of thiyl radicals but effectively slowed down the oxidative destruction of beta carotene by peroxynitrite and free radicals of lipids. In the presence of glutathione, the inversion of the antioxidant properties of DNIC into prooxidant ones took place. S-nitrosoglutathione prevented this inversion and suppressed the free-radical oxidation of beta-carotene induced by ferritin. It was proposed that the equilibrium between S-nitrosoglutathione, DNIC, "free Fe" ions and ferritin may determine the balance between prooxidant and antioxidant processes in living organisms. PMID- 12630108 TI - [Hydrodynamic parameters of native C-reactive protein molecule in a solution]. AB - The hydrodynamic properties of the C-reactive protein in solution (pH 6.8) were studied using quasi-elastic light scattering and size-exclusion liquid chromatography. It was shown that the solution containing the C-reactive protein represents a polydisperse system. The values of the translation diffusion coefficient and the apparent molecular weight of the C-reactive protein in solution at pH 6.8 were determined. The values of the translation diffusion coefficient, molecular weight and the hydration radius obtained suggest that the native pentameric C-reactive protein is the major form of the protein in solution at pH 6.8. PMID- 12630109 TI - [Irradiation by electrons of tubulin solution in vitro causes tubulin assembly in absence of a magnesium cofactor]. AB - It was shown that irradiation of a tubulin solution (5 x 10(-6) mol/l) with electrons makes tubulin assemble to microtubules. The response of the system was monitored by the femtosecond laser technique. The assembly under these conditions occurs without Mg2+ (magnesium cofactor) and GTP. At 730 (the first harmonic) and 365 nm (the second harmonic), a rise in signal intensity occurs during the first 60-70 ns followed by the onset of tubulin assembly to microtubules, which was registered by the methods of spectrophotometry and electron spectroscopy. Theoretically this effect can be explained by the appearance of hydrated electrons in the solvated tubulin shell. Hydrated electrons are mostly in a long living polaron state, which can be considered as an ensemble of quasi-dipoles of the electron-H3O+ type. The interaction of quasi-dipoles with the weak internal electrostatic field of tubulin leads, due to nonlinear effects, to a manifold rise in the intensity of the electrostatic field of the solvated shell of initially nonpolymerized tubulin chains. Finally, the increased field makes separate tubulin chains aggregate to microtubules. The effect observed is identical to the action of Mg2+ on the GTP site of beta-tubulin, which transfers it to a slightly perturbed triplet state. PMID- 12630111 TI - [The propagation of acoustic waves along a DNA molecule in viscous environment]. AB - Low-frequency vibrations of the structural elements of the DNA molecule in a viscous solution were investigated. It was shown that shear, twist and extension waves can propagate along the axis of the molecule. The mutual influence of acoustic waves on each other was analyzed. Formulas for determining the sound velocity and the attenuation coefficients for these types of waves are obtained. PMID- 12630110 TI - [Analysis of exciton parameters in DNA. Exciton waves in DNA as one of the reasons of mutagenesis]. AB - Formulae were obtained for the quantitative analysis of the following parameters of excitons in DNA: 1) the lifetime of electronic excitation; 2) the numbers of exciton runs along the DNA sequence; 3) the energy loss by an exciton for one run; 4) the maximum length of the DNA sequence capable of deactivating an exciton for one run. The maximum and minimum ranges for the constant of electronic excitation migration was determined to meet the requirement of inductive resonance energy transfer for the case of strong interaction. The constant of exciton energy migration was shown to depend on the activation energy, which is equal to the energy of absorbed quantum. An analytical formula was derived to determine the number of quanta the DNA molecule is able to absorb, depending on its length, without nonlinear effects (without overlapping of spatial areas of electronic excitation). By this formula, DNA sequences consisting of only identical AT and GC nucleotide pairs and aggregate AT + GC (in the ratio 1:1) DNA sequences ranging from 1 up to 10(10) base pairs were analyzed. The results of the analysis suggest that the overlapping of spatial areas of electronic excitation induced by a single ultraviolet quantum occurs in short DNA sequences characteristic of prokaryotes. To achieve the same effects on long DNA sequences specific for eukaryotes, DNA must synchronously absorb a great number of ultraviolet quanta. Based on the above results, the following conclusions were made: 1) disturbances in the normal activity of DNA and RNA polymerases may be due to electromagnetic field, which is caused by the oscillatory relaxation of vibronic levels of nucleotides. The energy enters the vibronic levels of nucleotides from an exciton running along the DNA sequence; 2) the increase in the noncoding DNA sequences in eukaryotes due to evolution is a way of DNA protection from undesirable mutations; 3) prokaryotes must possess a greater potentiality and a higher rate of mutagenesis in comparison with eukaryotes, which is proved by their greater diversity in nature. PMID- 12630112 TI - [Two-level heterogenous energy migration. Modeling and application to purple bacteria]. AB - The dynamics of migration of electronic excitations and the efficiency of their trapping in two-dimensional ensembles of molecules were analyzed. Molecules were characterized using the following parameters: the width of long-wavelength bands, the values of extinction and rate constant of deactivation of electronic excitations, critical distances of migration close to those of dye molecules, in particular, bacteriochlorophyll a and purple bacteria. A comparative analysis of two-dimensional models of energy migration made it possible to chose a model with an optimum light-harvesting on traps from the largest numbers of light-absorbing molecules. It was shown that in ensembles of molecules having different spectral characteristics (spectral shifts between the short- and long-wavelength fractions of the molecules are hear 800 cm-1) the efficiency of excitation trapping is approximately 90 and 80% for the number of light-harvesting molecules per one trap 210 and 580, respectively. PMID- 12630113 TI - [Thermodynamic parameters of free oxidation pathways in liver mitochondria]. AB - Two pathways of free oxidation in liver mitochondria were examined. One of these pathways is determined by the protonophoric action of free fatty acids, and the other pathway, by passive proton leakage in the absence of fatty acids. According to the model of the proton futile cycle of mitochondria, the protonophoric activity of fatty acids was defined as a quotient of the division of the acceleration of respiration by fatty acid by the coefficient of respiration control for the proton leakage. The temperature dependence of the palmitate protonophoric activity on the Arrhenius plot has a break at 22 degrees C and is characterized by the transition of activation energy from 120 to 60 kJ/mol. The dependence of the respiration rate in state 4 on the Arrhenius plot is linear and, the activation energy is 17 kJ/mol. It was concluded that the first pathway of free oxidation is determined by the cyclic transport of fatty acids with the participation of metabolic carriers, and this process depends on the membrane fluidity; the second pathway is determined by passive leakage of protons through membrane channels, without fatty acids and this process is independent on membrane fluidity. PMID- 12630114 TI - [Effect of redox-modified fibrinogen on blood leukocyte functions]. AB - The influence of oxidized fibrinogen on the intensity of luminol-dependent chemilumin escence of blood leukocytes, stimulated by opsonized zymosan was studied. It was shown that the introduction of fibrinogen modified by UV irradiation in to a suspension of cells resulted in a significant increase in the intensity of the luminol-dependent chemiluminescence of leukocytes. It was suggested that oxidized fibrinogen can influence blood leukocytes, enhancing their functional activity. PMID- 12630115 TI - [Selectivity of accumulation of chlorine e6 derivatives in blood leukocytes]. AB - The uptake of chlorine e6 derivatives by peripheral blood leukocytes was studied using a fluorescence-activated sorter. The analysis showed that the order of pigment uptake by leukocyte populations is the following: granulocytes > or = monocytes > lymphocytes. It was found that the accumulation of the pigments in the cell significantly varied. The level of chlorine e6 dimethyl ester accumulated by cells was found to be 15 times higher than that of chlorine e6. It was assumed that the differences in pigment uptake by different types of blood cells are due to structural and morphological features of leukocytes. The data obtained may be useful in developing the new methods of photodynamic therapy. PMID- 12630116 TI - [Effect of La(3+) on electrophoretic mobility and aggregation of intact human erythrocytes and those treated with low concentrations of glutaric aldehyde]. AB - It was shown that the treatment of erythrocytes with low concentrations of glutaraldehyde (0.01-0.1%) for 30-120 min to a variable extent the aggregation induced by 40-330 microM La3+. The effect of glutaraldehyde on the aggregation increased with concentration and time of fixation. La3+ ions decreased to a similar extent the electrophoretic mobility of intact erythrocytes and erythrocytes treated with 0.1% glutaraldehyde. No relationship was found between the change in the negative charge on the erythrocytes and the degree of their aggregation. Neuraminidase and trypsin were shown to decrease the surface charge on the erythrocytes and the aggregation of fixed erythrocytes. PMID- 12630117 TI - [Study of oxygenation of erythrocyte hemoglobin on shift and width of 1H NMR signal in blood water]. AB - It was found that the resonance frequency of NMR protons of the fluid inside erythrocytes differs from that of plasma protons. This leads to a broadening of the NMR signal from blood, which depends on the extent of oxygenation and the shape of erythrocytes. The coefficient of diffusion of water through the erythrocyte membrane was estimated to be D approximately equal to 10(-14) m2/s. PMID- 12630119 TI - [Treatment of burn surfaces by proteinases: mathematical description of an enzyme distribution]. AB - The process of penetration of a proteolytic enzyme applied to the surface of burn wound into the depth of necrotic tissue was considered. The model approximation describes three factors by a series of mathematical equations: inward-directed enzyme diffusion, counter-flow filtration of interstitial fluid (exudates), and irreversible inactivation of the enzyme by specific inhibitors present in exudates. According to the model, a quasi-stationary distribution of enzymatic activity through the thickness of the necrotic layer is achieved within 3 h and persists as long as the enzyme concentration on the wound surface is constant. The enzyme activity diminishes linearly from the wound surface to the mid-part of the necrotic layer. No enzyme activity is retained in the inner mid-part of the necrotic layer completely protected by the prevalent inhibitor. The ratio of enzyme concentration on the wound surface to inhibitor concentration in the interstitial fluid is the same as the ratio of the depth of active enzyme area to the depth of the inhibitor-protected area through the necrotic layer. The dynamics of accumulation of the active enzyme in the necrotic zone and the rate of enzyme inactivation in the wound by inhibitors were described by formulas applicable for practical purposes. PMID- 12630118 TI - [Study of water exchange in skin tissues by microwave dielectrometry]. AB - The complex dielectric permitivity of human skin was measured at a frequency of 42 GHz, and a method for calculating water content in skin tissues in vivo was proposed. The water content reaction of skin to standard physical exercises and nontoxic doses of nicotine was investigated. The functionally related changes in skin dielectric properties were interpreted as structural rearrangements of water contained in blood and tissues fluids. PMID- 12630120 TI - [Mathematical model of proliferative activity in epidermis of the normal skin and of the skin afflicted by psoriasis]. AB - A mathematical model of the mitotic activity of epidermis in norm and psoriasis is presented, which consists of a system of two autonomous nonlinear differential equations. A qualitative analysis of the model was done, and numerical solutions at the parameter values corresponding to these state were obtained. It was shown that, in norm, the system can exist only in one stationary equilibrium state of the "focus" type, and in psoriasis, due to an increase in the growing fraction, hyperproliferation, and enhanced migration of keratinocytes, a stable limiting cycle arises from the state of an unstable focus. The existence of two stable states (focus and the limiting cycle) is regulated by a parameter that describes the inhibition of division of maturing cells of suprabasal epidermal layers by the intrinsic tissue-specific transmitters of mitosis of G1-keilon type. The model is consistent with experimental data on the kinetics of cell proliferation in the epidermis in norm and psoriasis and the clinical course of the disease. PMID- 12630121 TI - [Mathematical modeling of non-stationary spatial gradients of nitric oxide in the muscle wall of blood vessels]. AB - A mathematical description of the diffusion of low-mass uncharged molecules in homogenous biological medium was considered. The distribution of nitric oxide in vascular smooth muscles was taken as an example. The model makes it possible to describe the spatial-temporal gradient of NO near the surface of endothelium in terms of one-dimensional diffusion. In the case of the constant rate of nitric oxide production in endothelium, an analytical expression of nonstationary distribution of NO concentration was obtained. It was found that, if the diameter of blood vessel exceeds 800 microns, the concentration of nitric oxide on the outer border of vascular smooth muscle layer is less than the equilibrium dissociation constant (0.25 microM) of soluble guanylyl cyclase. Thus, the diffusion of nitric oxide produced in endothelium is insufficient to cause a relaxation of vascular smooth muscles in medium- and large-size blood vessels. In this case, the role of nitric oxide synthesized in vasa vasorum acquires a special significance for the regulation of vessel tone. PMID- 12630122 TI - [Increased generation of hydroxyl radicals in the rat hypertrophied myocardium: in vivo study by microdialysis]. AB - A comparative study of the generation of hydroxyl radicals (OH*) in the hypertrophic myocardium of SHR-SP rats (n = 8) and in the myocardium of WKY (n = 5) and Wistar (n = 12) rats was performed using the microdialysis technique. The experiments were carried out on anesthetized open-chest male rats (ketamine intraperitoneally, 10 mg/kg) with artificial ventilation. The amount of OH* produced was estimated by high-performance liquid chromatography with electrochemical detection using as a marker 2,3-dihydroxybenzoic acid (2,3-DHBA), a product of the reaction of the hydroxyl radical with salicylic acid added to the perfusate. The quantity of 2,3-DHBA in the dialysate was estimated by the external standard method and expressed in percent of the 2,3-DHBA concentration in the perfusion fluid. The mean baseline value of 2,3-DHBA in dialysate samples in SHR-SP rats (157 +/- 22%, n = 8) was significantly higher than in Wistar (90 +/- 15%, n = 12, p = 0.0001) and Wistar-Kyoto rats (106 +/- 12%, n = 5, p = 0.005). The basal 2,3-DHBA level in SHR-SP rats was positively correlated (r = 0.831, n = 7, p < 0.05) with the degree of hypertrophy of the left ventricle expressed as the ratio of the left ventricle weight to the body weight. The data presented demonstrate that the hypertrophy of the left ventricle in SHR-SP rats is accompanied by the elevation of the level of free oxygen radicals. PMID- 12630123 TI - [Interdependence of plankton spatial distribution and plancton biomass temporal oscillations: mathematical simulation]. AB - The dynamics of aquatic biological communities in a patchy environment is of great interest in respect to interrelations between phenomena at various spatial and time scales. To study the complex plankton dynamics in relation to variations of such a biologically essential parameter as the fish predation rate, we use a simple reaction-diffusion model of trophic interactions between phytoplankton, zooplankton, and fish. We suggest that plankton is distributed between two habitats one of which is fish-free due to hydrological inhomogeneity, while the other is fish-populated. We show that temporal variations in the fish predation rate do not violate the strong correspondence between the character of spatial distribution of plankton and changes of plankton biomass in time: regular temporal oscillations of plankton biomass correspond to large-scale plankton patches, while chaotic oscillations correspond to small-scale plankton patterns. As in the case of the constant fish predation rate, the chaotic plankton dynamics is characterized by coexistence of the chaotic attractor and limit cycle. PMID- 12630125 TI - [Characteristics of the pressure center movement under upright posture regulation]. AB - The hypothesis was put forward that, along with the regulation of mass center projection, the system of upright posture control stabilizes the deviation of pressure center from the position of the mass center projection. The regularities in the behavior of the trajectories of pressure center and mass center projection were analysed. Experimental evidence was obtained supporting the validity of the hypothesis. The structure of the control system that corresponds to the new understanding of the variables being regulated during the maintenance of vertical posture was considered. PMID- 12630124 TI - [Effect of weak electromagnetic radiation on larva development and metamorphosis of grain beetle Tenebrio molitor]. AB - The effect of weak electromagnetic radiation (36 GHz, 100 mu W/cm2) on the development of the grain beetle Tenebrio molitor was studied. Insects were irradiated in different larval instars and at the pupal stage. It was found that weak electromagnetic radiation stimulated the molting and pupation of larvae and the metamorphosis of pupae. The stimulating effect of radiation was weak when animals were exposed in the initial period of the instar and the pupal stage and was more pronounced if the irradiation was carried out in the second half of the current instar and the pupil stage. The effect of weak electromagnetic radiation on the development of beetle can be related to the function of the hormones of metamorphosis. PMID- 12630126 TI - [Physical model of the plant tissue response to exposure to the microwave electromagnetic field]. AB - A hypothesis was suggested to explain the effect of biostimulation of seeds exposed to microwave electromagnetic field. It was shown that the assumption on the determining influence of the microwave field on the transport properties of the conducting system of a plant satisfactorily explains the phenomena observed in germinating seeds and growing plants. A physical model of the response of a plant cell to a microwave field is presented, which served as a basis for the method of calculating the maximum possible time of exposure of plant tissue. PMID- 12630127 TI - [Mathematical model of children mortality]. AB - A mathematical model of infantile mortality is proposed. The model is based on the probability principle of organism-environment interactions assuming that the organism is able to remember the diseases encountered previously and resist them. The adequacy of the model was assessed using the demographic database for two countries. The dynamics of the model parameters during the last century is presented. PMID- 12630128 TI - Keeping mental health reform on course: selecting indicators of mental health system performance. AB - Mental health reform was introduced as a public-sector policy response to identified deficiencies in the mental health services system. The reform agenda, which is underway in most provinces/territories, calls for greater accountability to mental health consumers, their families, and other stakeholders. This paper summarizes the major policy goals (which are the cornerstones of mental health reform) and suggests a series of high-level indicators to assess performance toward achieving these goals. Issues in the selection and measurement of performance indicators as markers of key goals are discussed. The proposed indicator set, while not a comprehensive framework, can provide a multi dimensional appraisal of mental health system performance and help keep reform on course. PMID- 12630129 TI - The Knowledge Resource Base: beginning the dialogue. AB - The Knowledge Resource Base (KRB) is a conceptual approach to the various types of knowledge used to understand and make sense of mental illness. It is an attempt to go beyond the everyday notion that real knowledge is in the hands of clinical experts and that consumers/survivors, families, and the public have little to contribute. There are four components to the KRB, each of which represents a different perspective or type of knowledge about mental illness; medical/clinical, social scientific, experiential, and customary/traditional. The purpose of this paper is to explore the medical/clinical and experiential components of the KRB by initiating a dialogue between consumers/survivors, families, and mental health professionals regarding these components. The strengths and weaknesses of each component are identified through individual interviews and a focus group. PMID- 12630130 TI - Childhood experiences with, and current attitudes toward, corporal punishment. AB - The purpose of the present study was to determine, in a Canadian sample, the extent to which corporal or physical punishment use continues, personal experiences, and current attitudes. Of the 436 participants, 75% reported receiving physical punishment as children. Approximately 40% of participants agreed that corporal punishment is necessary as a means of discipline. Since parental attitude toward physical punishment has been determined to be an important predictor in its use with children, the authors recommend that parent education programming must include information related to its risks. PMID- 12630131 TI - [Inner city children's perceptions about well-being]. AB - This study, carried out from the point of view of positive psychology, gave inner city children an opportunity to express themselves. Through interviews, they shared their perceptions of psychological well-being in the context of their everyday life and the contribution of a community agency towards their well being. The support of others appeared as a fundamental aspect of their perceptions of psychological well-being, while the enrichment provided by the agency also emerged from the data. However, children were shown to have relatively few personal strategies to improve their own well-being. The children also identified some obstacles to their well-being. A series of tailored reporting activities made it possible for the personnel and clients of the agency to appropriate the results of the study. PMID- 12630132 TI - The long-term impacts of group treatment for partner abuse. AB - This paper evaluates Responsible Choices for Men, a 14-week therapy group for abusive men based on feminist perspectives using narrative methods. Pre-test and post-test information was available for 76 program completers measuring physical and non-physical abuse, self-esteem, perceived stress, family relations, depression, assertiveness, and sex-role beliefs. The men significantly improved on all variables. A further focus was following up with 22 group members, contacted 5 to 28 months post treatment and asked to complete the same measures. The results indicated not only maintenance of the post-group changes, but continued improvement. Clinical implications and recommendations for group programming with abusive men are presented. PMID- 12630133 TI - [Screening protocol of spouse abuse. Efficacy measure]. AB - Screening protocols that can be used to identify domestic violence are lacking (Gendron, 1987), as are studies evaluating the effects of using these protocols. With the objective of screening domestic violence, the Saint-Hubert CLSC has developed a protocol by in collaboration with the Centre de recherche interdisciplinaire sur la violence familiale et la violence faite aux femmes (CRI VIFF). Evaluation of the domestic violence screening protocol established at the Saint-Hubert CLSC indicates that the number of cases in which domestic violence was detected was 3 times higher after the screening protocol was put in place, and 15 times higher at the Saint-Hubert CLSC than at the CLSC used as a comparison site. PMID- 12630134 TI - Claiming our place: women with serious mental health issues and support groups for abused women. AB - Many women with serious mental health issues also deal with abuse and have difficulty accessing services. Despite the fact that groups have been found to be one of the most useful tools in healing from the effects of abuse, many professionals see women with serious mental health issues as unable to benefit from counselling and, in particular, from groups for abused women. This study indicates that, when mental health issues are addressed and the group structures and expectations are modified to allow women control over their participation, serious mental health issues are not a barrier to participation in groups. PMID- 12630135 TI - [Quality and conditions of the collaboration with the families viewed by the personnel working with psychiatric and traumatic brain injury clients]. AB - This paper describes the point of view of professionals working with traumatic brain injury (TBI) clients or people with schizophrenic disorders towards their collaboration with clients' families. Four hundred and thirty professionals from 3 different regions in the province of Quebec took part in the study. They were recruited from psychiatric and general hospitals and physical rehabilitation centres. The results of the study point up that conditions for collaboration with families are better for respondents working with TBI clients than for respondents in the psychiatric sector. Furthermore, the results showed that respondents working with TBI clients are significantly more satisfied with their working conditions and the services they deliver and perceive a higher degree of collaboration with families than mental health workers. Another finding of the study is that personnel intervening in an interdisciplinary team reported a higher degree of collaboration with families than respondents working in a multidisciplinary team setting or individually. In the same vein, respondents who received training regarding intervention with families during their practice or during their academic training reported a higher degree of collaboration with their clients' families. PMID- 12630136 TI - Supported housing for people with serious mental illness: resident perspectives on housing. AB - Supported housing involves integrated housing that is adequate and affordable, paired with flexible, individualized mental health support services. In this qualitative study, interviews were conducted with supported housing residents in cities and towns in southwestern Ontario to examine their housing experience. Questions were organized around 4 dimensions of housing: (a) physical environment, (b) social environment, (c) affordability and choice, and (d) residential history. The inquiry, which occurred at neighbourhood and dwelling unit levels, revealed 4 themes: (a) loneliness, (b) making do with socially and structurally inferior housing, (c) a desire for more understanding, and (d) a concern with an individual's sense of integration into a community. PMID- 12630137 TI - [Attitudes and beliefs of citizens of Montreal towards psychiatric patients living in a society]. AB - In the past 50 years, new health policies all over the world have led to the introduction of rehabilitation and deinstitutionalization programs to overcome the depersonalizing effects of asylums. As a result, large numbers of former psychiatric patients are now living and working in society. The public's attitudes and beliefs about these former patients play a central role in their harmonious integration into the community. To find out more about these attitudes and beliefs, we interviewed a representative sample of the population of Montreal Island (n = 899). Our study indicates that a majority of respondents show a favourable attitude towards former psychiatric patients (in terms of recognizing their rights, accepting them in the workplace, creating warm relationships, etc.), but that people still entertain many negative beliefs (seeing mental illness as a deviation from the norm). The study interprets these ambivalent signs of tolerance and fear towards ex-psychiatric patients among Montrealers. An encouraging sign is that a trend towards greater acceptance seems to be emerging among young people in terms of both attitudes and beliefs. PMID- 12630138 TI - Principles of highly effective pain managers. PMID- 12630139 TI - Integrative care--Reiki. PMID- 12630140 TI - MMR--a good news story. PMID- 12630141 TI - New maternity and paternity rights: 2003. PMID- 12630142 TI - Diarrhoea and gastroenteritis in the infant and young child. AB - Gastroenteritis is a major cause of diarrhoea in children. These children first present in the primary care setting for management, and the management and advice given varies between different practitioners. Unfortunately, the treatment and advice offered can occasionally be inadequate and inappropriate. This article highlights basic principles of management of gastroenteritis in children. The aim is to produce easily understandable, usable and reproducible evidence-based guidelines. PMID- 12630143 TI - MMR vaccination: answers to some parents' questions. AB - Following adverse publicity about the measles, mumps and rubella (MMR) vaccine, most parents ask their health visitors, GPs and practice nurses about the risks and benefits. To address parents' concerns and help health professionals answer their questions, the Department of Health has provided information through its agency NHS Immunisation Information. This article contains some of the commonly asked questions together with answers and references. PMID- 12630144 TI - Adoption--3: The adopted child at school. AB - In the last of three articles, Patricia Swanton explains some of the particular needs an adopted child may have because of present circumstances and past trauma. She discusses ways in which teachers and health professionals can best support the adoptive parents and their child so that the child is able to make the most of being at school. PMID- 12630145 TI - Putting people first. PMID- 12630146 TI - Issues in infant and toddler nutrition. AB - In 2002 SMA Nutrition organised several regional study days where expert speakers discussed issues including childhood obesity, under-nutrition, gastro-oesophageal reflux and the importance of essential nutrients. PMID- 12630147 TI - Maternal nutrition and low birth weight. AB - The UK has the same incidence of low birth weight babies as Romania--7% of all births. The incidence is higher in deprived areas. In Hackney, east London, 10% of babies are below 2.5 kg at birth. Infant mortality is higher in smaller babies and they are more likely to be sickly infants, have congenital abnormalities or under-achieve at school. PMID- 12630148 TI - Long chain polyunsaturated fatty acid levels in chronic illness. AB - Abnormal long chain polyunsaturated fatty acid (LCP) levels are found in children and adults who have several chronic diseases, due to restricted diets or possible abnormalities in metabolic pathways. In some cases, LCP supplementation is effective. Judy More looked at studies assessing LCP levels and supplementation in children with chronic diseases. PMID- 12630149 TI - Identifying and managing faltering growth. AB - Five per cent of under-5s will have an episode of faltering growth. 'We need to identify and support these children to prevent problems later' said Sue Barraclough. If unchecked, faltering growth can lead to stunting and delayed development. Many factors contribute to growth faltering, as shown in Figure C. Signs to look out for are listed in Table A. PMID- 12630150 TI - Long chain polyunsaturated fatty acids improve cognitive development. AB - Docosahexaenoic acid (DHA, an Omega-3 fatty acid) and arachindonic acid (AA, an Omega-6 fatty acid) are long-chain polyunsaturated fatty acids (LCPs) that are important for cognitive development. Research has shown that LCPs are associated with improved visual and cognitive development: breast-fed children had higher IQ scores compared with children who received an infant formula that did not contain LCPs. Because breast milk contains LCPs and the formulae in these studies did not, it is possible that LCPs may contribute to improved cognitive development. PMID- 12630152 TI - [Aging and Nursing Care Specialty Congress with a new image: nursing care with more prestige]. PMID- 12630151 TI - Reflux: causes and treatments. AB - Although gastro-oesophageal reflux (GOR) symptoms diminish in 80% of babies by their first birthday, this common problem causes parents and babies much distress. In extreme cases, GOR is linked to oesophageal ulceration and faltering growth. But most infants have uncomplicated GOR without a definitive cause. GOR symptoms can resolve using agents or special infant formulae that thicken the gastric contents. It is important to try and avoid invasive procedures in children with GOR. PMID- 12630153 TI - [Otorhinolaryngology--2: "Finger tip sense" is needed]. PMID- 12630154 TI - [Introduction to nursing diagnosis: general practice experiences in Austria]. PMID- 12630155 TI - [Nurse counseling in the Munich-Neuperlach Hospital: initial results of a model project]. PMID- 12630156 TI - [Organization of a center for nursing data and information: a pilot project for nursing documentation]. PMID- 12630157 TI - [Introduction of computer-assisted nursing documentation: quality improvement with PIK (Nursing Information and Communication System)]. PMID- 12630158 TI - [A concept of nursing care relations: the roles play a role]. PMID- 12630159 TI - [Autonomy and private sphere in hospitals: patients often have a different concept from nurses]. PMID- 12630160 TI - [Education in nursing care: recognition is still missing (interview by Stephanie Weiss and Katrin Balzer)]. PMID- 12630161 TI - [Personal development in nursing--II: Growth through more responsibility]. PMID- 12630162 TI - [Preparations in personal capital financing--3: Are your accounting records and controlling fit for rating?]. PMID- 12630163 TI - [Telephone during work schedule? Nurse have a right to undisturbed leisure time]. PMID- 12630164 TI - [Learning in nursing care: air for life]. PMID- 12630165 TI - [Modern hospice movement in Germany--III: From institutional resistance to acceptance]. PMID- 12630166 TI - [An occupation "conquers" the world. Advanced nursing practice]. PMID- 12630167 TI - [MEDICA 2001 in Dusseldorf: money, markets, possibilities]. PMID- 12630168 TI - [Otorhinolaryngology--3: Regulations for managing hearing aids]. PMID- 12630169 TI - [Giving birth in the maternity house. Where trust is at home]. PMID- 12630170 TI - [Nursing care for immigrants: ethnic-cultural aspects in maternity care]. PMID- 12630171 TI - [Nutrition of pregnant and puerperal patients in other cultures: 4 women and the special power of soup]. PMID- 12630172 TI - [Bonding: the most important bond in life]. PMID- 12630173 TI - [The very small premature infant: trust in the will to survive]. PMID- 12630174 TI - [With experience grows acceptance: advantages and disadvantages of binasal CPAP ventilation]. PMID- 12630175 TI - [Management of women with postpartum depression: a project for return to happiness (interview by Katrin Balzer)]. PMID- 12630177 TI - [Autonomy of postpartum women: different viewpoints of mothers and nurses]. PMID- 12630176 TI - [Applying new forms of nursing care in the obstetrics department: quality of graduate and continuing education determines success]. PMID- 12630178 TI - [Learning in nursing care: a disaster seldom comes by itself]. PMID- 12630179 TI - [Preparations for personal capital financing--4: Is your marketing fit for a rating?]. PMID- 12630180 TI - [Nursing care for the elderly: professional ethics in view of the new regulations]. PMID- 12630181 TI - [Autonomy, privacy and informed consent: a BIOMED II Research Project with patients and nursing personnel in 5 European countries]. PMID- 12630182 TI - ST-segment elevation myocardial infarction, severe anemia and nonobstructive coronary disease: case report and brief comment. AB - When the metabolic demand of the heart exceeds its oxygen supply, myocardial ischemia or infarction (MI) may occur. One of the causes of this condition is severe anemia. The type of MI associated with cases of supply-demand myocardial mismatch is usually a non-ST elevation infarction (Non-STEMI), and is most commonly seen in patients with underlying coronary artery disease. We present a case report of ST segment elevation MI (STEMI), which we believe was secondary to severe anemia, occurring in the absence of angiographically significant coronary artery stenosis or thrombosis. Coronary artery spasm was not seen. In our review of the literature we found no similar cases (MEDLINE 1966-2002). PMID- 12630183 TI - State mandated prenatal human immunodeficiency virus screening at a large community hospital. AB - PURPOSE: To describe the initial experience of state mandated prenatal HIV screening at a large community hospital. METHODS: HIV screening was provided to all pregnant women as of October 1, 1999. All HIV-positive women identified received aggressive antiretroviral therapy to reduce the likelihood for vertical transmission. Neonates were screened for HIV at zero, six, and 12 months of age. RESULTS: Seven pregnant women (0.3%) and two additional family members tested positive for HIV. All seven infants born to the identified HIV-positive women have tested negative for infection. We estimated that six of nine cases of HIV infection identified would have been missed under a policy of voluntary HIV screening. CONCLUSIONS: Universal screening for HIV in pregnancy is achievable and desirable and provides the best opportunity to minimize the number of new neonatal HIV infections. PMID- 12630185 TI - How do we keep going? Personal and professional meaning after 9-11. PMID- 12630186 TI - The cost of health care. PMID- 12630187 TI - End-of life care. PMID- 12630184 TI - Use of activated protein C (drotrecogin alfa) in a patient with sepsis and respiratory failure on ultra high frequency jet ventilation. AB - Drotreocogin alfa is a recombinant form of human activated protein C that has recently been found to reduce mortality significantly when used in patients with severe sepsis. Bleeding is reported to be the most common adverse effect associated with the use of this drug. Patients with sepsis on Ultra High Frequency Jet Ventilator may develop necrotizing tracheobronchitis and may be at an increased risk of bleeding when treated with drotreocogin alfa. We describe a patient with sepsis and respiratory failure on Ultra High Frequency Jet Ventilator, who was started on drotrecogin alfa, without the development of any significant bleeding. PMID- 12630188 TI - Re: Management of cancer-related and non-cancer-related chronic pain in Connecticut: successes and failures. PMID- 12630189 TI - [Improving child health. Interview by Catherine Maisonneuve]. PMID- 12630193 TI - [School mediation as a symptom of staff division in schools]. PMID- 12630194 TI - [Nursing, a profession in student relations]. PMID- 12630195 TI - [School nursing, a profession with many unknown facets]. PMID- 12630197 TI - [Health and preventive care in students in preparatory classes]. PMID- 12630196 TI - [School health, a challenge to succeed with students, a constant preoccupation for school nurses]. PMID- 12630198 TI - [Sexual education. Talk to me of love, tell me tender things...]. PMID- 12630199 TI - [Influence of cannabis use on the school work of adolescents]. PMID- 12630200 TI - [Parties for hospitalized children]. PMID- 12630201 TI - [Emergencies. 4/7 Atresia of the small intestine]. PMID- 12630202 TI - [Shock--3: Forms of shock: etiology determines therapy]. PMID- 12630203 TI - [Modern therapy of breast carcinoma: more time for living]. PMID- 12630204 TI - [Systemic whole body hyperthermia in oncology: fatal heat for tumor cells]. PMID- 12630205 TI - [Prevention and management of lymphedema: increasingly fewer women are at risk]. PMID- 12630206 TI - [Incontinence in the woman--forms and management: more attention to a taboo]. PMID- 12630207 TI - ["Primary nursing": everything from the same team]. PMID- 12630208 TI - [Life after breast cancer: taking destiny into one's own hands (interview by Katrin Balzer)]. PMID- 12630209 TI - [Fatigue in breast cancer patients with adjuvant chemotherapy: the 2nd day is the worst]. PMID- 12630210 TI - [Nursing care of the umbilical cord: health measures have first priority]. PMID- 12630211 TI - [Quality assurance in promoting breast feeding: the WHO/UNICEF "Supportive Breast Feeding Hospital Environment" Initiative]. PMID- 12630212 TI - [Quality assurance in nursing insurance: improvements not always for the better]. PMID- 12630213 TI - [Nursing home quality assurance regulation: using the chance for a new beginning]. PMID- 12630214 TI - [Opposition in National Socialism: the saving angel]. PMID- 12630215 TI - [A story: spitting]. PMID- 12630216 TI - [From case to case: "Young lady, young lady"]. PMID- 12630217 TI - [Wound management--1: Regular water--safe for wound cleansing?]. PMID- 12630218 TI - [Concept for assessment and evaluation of quality in ambulatory nursing services]. PMID- 12630223 TI - [Thirty five hours in practice. Mitigated success]. PMID- 12630225 TI - [Description and function of sterile isolators]. PMID- 12630224 TI - [Treatment of hereditary blood diseases by hematopoietic stem cell allografts]. PMID- 12630226 TI - [Chloe's day or nursing care in a sterile isolator]. PMID- 12630227 TI - [Psychological repercussions in bone marrow transplantations]. PMID- 12630228 TI - [Children in sterile isolators: which social management of the family?]. PMID- 12630229 TI - [Physical therapy-massage of children in a sterile isolator]. PMID- 12630230 TI - [Educational management and music therapy of children in sterile isolators]. PMID- 12630231 TI - [Diploma state nurses and smoking cessation in nursing care]. PMID- 12630233 TI - [Best practice guidelines in stomatherapy]. PMID- 12630232 TI - [A new therapeutic alternative in chronic B-lymphocytic leukemia]. PMID- 12630234 TI - [Clinical audit of blood pressure measurement by third year students]. PMID- 12630236 TI - [To the aid of homeless persons]. PMID- 12630235 TI - [Carbamazepine]. PMID- 12630237 TI - [A student chooses community health nursing]. PMID- 12630238 TI - People with dementia deserve free care. PMID- 12630239 TI - A nursing contribution to research issues in blood transfusion. AB - Research nurses at the National Blood Service are working on projects as diverse as identification barcodes, a national profile of blood recipients and pre operative transfusion in sickle cell disease. PMID- 12630240 TI - Tackling childhood obesity. AB - The number of overweight or obese children has increased markedly over the past 20 years in the UK. How can health-care staff best tackle sedentary lifestyles and poor eating habits? PMID- 12630241 TI - Insulin-delivery devices. AB - People with diabetes requiring insulin now have a range of insulin-delivery devices, the use of which can help to ensure concordance with treatment. PMID- 12630242 TI - How a specialist nurse helps patients undergoing deep brain stimulation. AB - Deep brain stimulation is becoming the most frequently used neurosurgical treatment for patients with complex Parkinson's disease. This paper reports on the creation of a clinical nurse specialist role in a Scottish hospital's movement disorder team to support patients going through this treatment and their families, working with the neuroscience nurses and multidisciplinary team. PMID- 12630243 TI - Cardiac care: 1. Interpretation of electrocardiogram rhythm strips. AB - This is the first in a series of three papers that aim to explain the anatomy and physiology related to cardiac conductivity and to describe some common and less common arrhythmias that ward nurses may come across. This first paper examines the background to ECG monitoring, the anatomy and physiology of the heart and ECG recording and interpretation. PMID- 12630244 TI - Informing the future: 2. Developing the role of community IPCNs. AB - This paper is the second in a two-part series summarising the main findings and conclusions of a review of the roles and responsibilities of infection prevention and control nurses commissioned by the Department of Health. It describes the core work of IPCNs, discusses the strengths and constraints of the current role and makes suggestions for future development. PMID- 12630245 TI - What to expect from an inquest and how to be prepared for difficulties. AB - An inquest aims to find out the facts surrounding a sudden death without apportioning blame. When a patient dies in hospital, if the cause of death is unknown or unnatural, nurses can be called to the inquest as witnesses. This paper explains how to prepare for this, what to expect during a hearing, and the importance of using the experience for reflective practice. PMID- 12630246 TI - Audit of wound swab sampling: why protocols could improve practice. AB - Many nurses carry out wound sampling using a surface swab, to establish infection levels and determine treatment. But reliability can be compromised by a number of factors. This paper examines reasons for sampling and its limitations. It describes how an audit of secondary and primary care nurses revealed wide variations in practice, and proposes protocols to improve care. PMID- 12630247 TI - Less is better: why venepuncture practice should be reviewed for those with end stage renal failure. AB - This paper proposes that replacing adult vacutainers with paediatric containers during the taking of blood samples would reduce the incidence of anaemia in patients with end-stage renal failure. PMID- 12630248 TI - Inadequate nursing care and the failure to keep adequate records. AB - Lessons can often by learned by studying the outcomes of complaints made to the NHS by patients. We look at three recent investigations conducted and published by the Health Service Ombudsman. The first and second relate to poor care and lack of communication, and the third to care of a patient's diabetes and epilepsy in hospital and failure to record an outpatient consultation. PMID- 12630249 TI - Safeguarding children: 2. Identifying and preventing institutional racism. AB - Racism can prevail as part of the ethos or culture of an organisation. Part two of this four-part Study paper on child protection discusses institutional racism within child-care services and the damage that can be caused by myths and stereotypes. Increasing the number of black and ethnic-minority staff within the NHS to reflect the communities they serve is an important step. PMID- 12630250 TI - Vitamins. PMID- 12630251 TI - HIPAA privacy regulations. PMID- 12630252 TI - Helping women with low libido. PMID- 12630253 TI - Preparing for practice: part of preparing for certification. PMID- 12630254 TI - Antibiotic sensitivity assays. PMID- 12630255 TI - And then there were six. Fewer states withhold CS authority from NPs. PMID- 12630257 TI - Patient information. The keys to weight loss: baby steps. PMID- 12630256 TI - Systemic lupus erythematosus. Early identification, co-management are key NP contributions. PMID- 12630258 TI - Growth hormone deficiency in children. A change in diagnostic approach. PMID- 12630259 TI - STD treatment update. A closer look at CDC guidelines. PMID- 12630260 TI - Home-based primary care for dementia. Project ahead. PMID- 12630261 TI - Balneo phototherapy for psoriasis. Modern application of an age-old treatment. PMID- 12630262 TI - A zebra you can't afford to miss. Wegener's granulomatosis. PMID- 12630263 TI - The other side of the stethoscope. Nurse practitioners as patients. PMID- 12630264 TI - Bone disease in HIV. What is the link? PMID- 12630265 TI - Question re legal column. PMID- 12630266 TI - The 10/90 gap. What it is, and what some Canadians are doing to close it. PMID- 12630267 TI - Early prevention. Occipital flattening of positional origin. PMID- 12630268 TI - Gerontology. Challenges in emergency room care for the elderly. PMID- 12630269 TI - Primary health care. Integrated services in northeast Edmonton. PMID- 12630270 TI - The meaning of youth participation. PMID- 12630271 TI - Introduction: moving youth participation forward. AB - Given the emerging interest among researchers, practitioners, and policymakers in youth participation, it is important to examine and assess carefully the promise and challenges of youth engagement. PMID- 12630272 TI - From assets to agents of change: social justice, organizing, and youth development. AB - Young people are joining together to demand a voice in the decisions that affect their lives. In the process, they are transforming policies and making institutions more accountable. PMID- 12630273 TI - Youth conferences as a context for engagement. AB - Youth conferences can be effective means for engaging youth in the life of their schools, communities, and nation, bringing benefits not only to youth themselves but also to the community, which gains through the energy, ideas, and values that youth contribute. PMID- 12630274 TI - Building young people's public lives: one foundation's strategy. AB - Foundations are awakening to the untapped potential of serious, policy-focused community change efforts led by teenagers and young adults. This chapter lays out background questions, a point of view, and programmatic strategies that one foundation developed for supporting young people who are taking direct action to improve their lives and communities. PMID- 12630275 TI - Moving youth participation into the classroom: students as allies. AB - The experiences of urban public high school students, told in their own words, offer new and veteran teachers guidance on how to reach adolescent learners and illustrate what youth-adult partnerships in the classroom might look like. PMID- 12630276 TI - Youth evaluating programs for youth: stories of Youth IMPACT. AB - Youth evaluators describe their findings from an extensive evaluation of forty youth programs in San Francisco. Interviews with current youth and the former program director provide insight into the promise and challenge of youth participation. PMID- 12630277 TI - Let's work together. Interview by Kathy Oxtoby. PMID- 12630278 TI - Myocardial infarction: signs symptoms and treatment. AB - From early middle age onwards myocardial infarction is recognised as being a major cause of death and disability in this country. Approximately 300,000 people in the UK experience an MI each year and about 140,000 will die as a result. The rapid management of patients with chest pain should ensure that there is no more than 30 minutes delay in decision-making and initiation of thrombolytic therapy. This 'door-to-needle' time should be reduced to 20 minutes from April 2003 in line with national service framework recommendations. This article provides an overview of the signs, symptoms and diagnosis of this acute condition. The contemporary management/treatment advocated in the NSF is presented and nursing interventions are explored briefly. PMID- 12630279 TI - What you need to know about...iron deficiency anemia. PMID- 12630280 TI - Red blood cell count. PMID- 12630281 TI - Sexual health in cancer care. AB - Two studies investigating nurses' involvement in supporting cancer patients with sexuality and fertility concerns were carried out after listening to the experiences of patients living with cancer. Findings from the studies revealed that while nurses recognised their role in supporting patients with sexual and fertility concerns, they avoided the subject, citing reasons such as lack of knowledge, experience and resources. The findings of one of the studies are reported here, together with the subsequent clinical practice developments that were initiated. PMID- 12630282 TI - The supply of antibiotics by NHS walk-in centre nurses using PGDs. AB - Patient group directions (PGDs) have become a feature of NHS walk-in centres enabling nurses to supply and administer drugs in response to the generalised direction of a doctor. In light of the issues surrounding the use of antibiotics a clinical audit was undertaken to assess nurses' competence and record-keeping in relation to supplying antibiotics according to PGDs. This study goes some way to demonstrate that nurses are supplying and administering antibiotics judiciously and safely according to local PGDs. PMID- 12630283 TI - 'It's emotionally draining but rewarding'. PMID- 12630284 TI - A breath of hope for the Big Apple. Interview by Leona Armstrong. PMID- 12630286 TI - Part-time workers' pension rights. PMID- 12630285 TI - Human approach to older people's care. Interview by Craig Kenny. PMID- 12630287 TI - Midwives taking on more roles: the 1st on call project. AB - On reflection I agree that midwives should not take on further roles if they cannot give one-to-one care in labour, which was the main point in the motion debated at the RCM conference last year. This is not the case at the Trust I work, and therefore extending our role feels correct and possible. I am undertaking a piece of research regarding the project, called 'blurring the boundaries', which hopefully will be completed soon. Role changes areas -- not just users. The NHS Plan implies health professional should be trying to work in partnership and I believe the 1st on call project has enhanced that process. PMID- 12630288 TI - Integrated approaches to therapeutic target gene discovery. AB - The identification of tractable drug targets is the first critical step in the long process of drug development, and the challenge for scientists in the post genomic era is to couple gene and protein sequence information with biological insight to identify genes with the greatest therapeutic and commercial potential. An extraordinary array of genomics- and proteomics-based techniques is available for this endeavor, and combining multiple, complementary approaches enhances the informative power of such experimentation. PMID- 12630289 TI - Aptamers and aptazymes: accelerating small molecule drug discovery. AB - Synthetic nucleic acid ligands, known as aptamers, are versatile tools that can greatly enhance the efficiency of modern drug development. Exhibiting binding characteristics comparable to or even better than monoclonal antibodies, these ligands can be used as detection probes, highly efficient inhibitors of protein function or specific competitors in high-throughput screening (HTS) assays. Thus, aptamer technology can be exploited to address the growing demand for multi parallel analysis of proteomes, functional prioritization of potential drug targets and accelerated small molecule lead identification. The unique advantages of this technology are the rapid automated generation of sophisticated ligands against almost any target molecule and the convenient structural or chemical modification of the nucleic acid probes. Depending on the strategy, an RNA aptamer can be expressed transgenically to investigate and inactivate an endogenous protein in an animal model, or it can be designed to function as a highly sensitive nucleic acid biosensor. More recently, the technology has been extended to directly link functional target validation with HTS, accelerating the process of drug discovery. PMID- 12630290 TI - Targeting protein kinases in cancer therapy. AB - Over the years, many different tyrosine and serine/threonine protein kinases have been selected as candidates for drug discovery activities in oncology research, based either on their overexpression and/or dysfunction in a particular organ or tissue, or through their association in deregulated signal transduction/cell cycle pathways. This review summarises current preclinical and clinical knowledge of ATP-competitive, small molecule kinase inhibitors, and receptor or ligand competitive antibodies of selected protein kinases in which drug discovery and development activities have advanced with some success. PMID- 12630291 TI - Kinases as targets: prospects for chronic therapy. AB - Several inhibitors of protein kinases have been launched or have reached late stages of clinical trials for the treatment of acute conditions such as cancer. Development was slow, mainly because of the associated toxicity, attributed to the poor selectivity of these compounds. Protein kinase inhibitors mainly bind at the active site of the enzyme, in competition with ATP, and whether such inhibitors could ever be used for the long-term treatment of chronic conditions, such as rheumatoid arthritis, is still questionable. This review concentrates on three enzymes, well established as potential drug targets for the treatment of chronic inflammatory conditions, and considers progress made in developing potent and selective inhibitors that could progress through clinical trials and eventually be launched on the market. PMID- 12630292 TI - Protein kinase inhibitors from the urea class. AB - Protein kinase inhibitors hold great potential as novel therapies for cancer and inflammatory disorders. While bis-aryl ureas have been reported as kinase inhibitors as early as 1996, a number of publications and patent applications appeared in the literature during the past two years. Three urea-based kinase inhibitors are currently undergoing clinical trials. The present review summarizes available data, and provides an overview of the structure-activity relationships against a variety of kinase targets, including p38, Raf-1 and cyclin-dependent kinases. PMID- 12630293 TI - Bioactive lipids and their receptors. AB - Recent studies show that several, if not all, degradative products of glycerol- and sphingosine-based lipids are active molecules that play important roles in different signal transduction pathways. The functions and actions of lysophosphatidic acid and sphingosine 1-phosphate, the degradation products of phospholipids and sphingolipids, respectively, have received a great deal of attention during the past few years. These lipid mediators are active through several abundant G protein-coupled receptors that were recently identified and characterized. This review will discuss recent achievements in the area of bioactive lipids. PMID- 12630294 TI - The inner workings of the AMPA receptors. AB - As with all ligand-gated ion channels, ionotropic glutamate receptors (iGluRs) convert the free energy of agonist-binding into a sequence of conformational changes that allow regulated ion flux across an otherwise insulating membrane bilayer. In the case of iGluRs, the resulting membrane depolarization is a key mediator of intercellular signals in the synapses of the CNS. Recent biomedical and structural research has greatly expanded our understanding of the chemistry of iGluR agonist binding, and of the subsequent molecular changes that drive channel gating and desensitization. PMID- 12630295 TI - Allosteric modulation of G protein-coupled receptors. AB - Allosteric modulation of G protein-coupled receptors has recently been recognized as an alternative approach for selectivity in drug action. Allosteric modulators that enhance or diminish the effects of (endogenous) agonists or antagonists on a variety of G protein-coupled receptors are described in this review, with emphasis on the latest developments in this research area. Specific examples include allosteric ligands for adenosine A1 and A3 receptors, Ca(2+)-sensing receptors, metabotropic glutamate receptor subtypes, gamma-aminobutyric acid type B and muscarinic receptors. It appears that all three major classes of G protein coupled receptors (A, B and C) are amenable to allosteric modulation by small molecules. This constitutes an attractive and novel means to identify new leads in the drug discovery process. However, it requires a reengineering of most current assays. PMID- 12630296 TI - Formation of oligomers by G protein-coupled receptors. AB - There is increasing evidence that G protein-coupled receptors (GPCRs) may form homo- or hetero-oligomers and that this may be important for their function. Evidence in favor of oligomerization comes from biochemical studies, studies of functional complementation and recent studies using energy transfer techniques, which provide direct evidence for receptor/receptor contacts. The oligomerization process may be regulated by ligands for some GPCRs or it may be constitutive in other cases. Homo-oligomerization may lead to cooperative behavior for the binding of ligands to some receptors. Hetero-oligomerization may generate new pharmacological properties for some receptor combinations as well as altering the signaling properties of the GPCR concerned. Additionally, the trafficking of GPCRs from the cell surface may be affected by oligomerization. PMID- 12630297 TI - Targeting neurotensin receptors with agonists and antagonists for therapeutic purposes. AB - Neurotensin (NT) is a brain-gut tridecapeptide that fulfils a dual function, as a neurotransmitter/neuromodulator in the nervous system, and as a paracrine and circulating hormone in the periphery. Three NT receptors, NTS1, NTS2 and NTS3, have been cloned to date. NTS1 and NTS2 belong to the family of G protein-coupled receptors with seven transmembrane domains, whereas NTS3 is a single transmembrane domain protein that belongs to a recently identified family of sorting receptors. Most of the known peripheral and central effects of NT are mediated through NTS1. NTS2 might take part in the analgesic response elicited by central administration of NT; the biological roles of NTS3 are yet to be discovered. Most NT agonists and non-peptide antagonists developed to date have been studied for their NTS1-targeting abilities. Here, we will discuss the potential diagnostic and therapeutic uses of these compounds in cancer, schizophrenia, obesity and pain suppression. PMID- 12630298 TI - Internal initiation of translation by viral and cellular IRESs--a new avenue for specific inhibition of protein synthesis? AB - Many virus species, as well as a limited number of cellular mRNAs, initiate protein synthesis by an unusual mechanism, based on well-defined RNA structures called internal ribosome entry sites (IRES). IRES-mediated internal initiation allows recognition of the start codon by the ribosome in a cap-independent way, avoiding major regulatory steps. The IRES elements and their trans-acting factors are potential targets for developing new agents against hepatitis C, and may provide new possibilities for antitumor therapy. PMID- 12630299 TI - Novel approaches to antimicrobial therapy: peptide deformylase. AB - Peptide deformylase (PDF) represents one of the most exciting new targets for the development of novel antimicrobial chemotherapies. PDF is an essential bacterial metalloenzyme that deformylates the N-formylmethionine of newly synthesized bacterial polypeptides. Recent progress in understanding the structure and function of PDF has greatly facilitated the drug discovery process. In this article, the potential of PDF as an antimicrobial target is reviewed, and progress in the development of PDF inhibitors (PDFIs) is highlighted. Several structural classes of compounds have been reported as inhibitors of PDF. However, the real challenge has been in obtaining molecules with potent in vivo antibacterial activity against a range of drug-resistant pathogens. One of the more encouraging compounds reported, BB-83698 (British Biotech plc/Genesoft Inc), has shown in vivo efficacy against Streptococcus pneumoniae in both mouse thigh and lung infection models at doses equivalent to existing therapies. The published data suggest that PDFIs are a promising new class of antimicrobial agent best suited to treat respiratory tract infections (RTIs), but with the potential for activity against a variety of other pathogens. It is anticipated that the first PDFI targeting RTIs will enter the clinic soon. PMID- 12630300 TI - New approaches to reversal of neuromuscular block. AB - Although numerous reversal agents for neuromuscular block (NMB) have been known for some time, investigations on new approaches were initiated only recently. The different approaches used in an attempt to avoid the muscarinic side effects associated with the antagonists of NMB that are currently available are reviewed. PMID- 12630301 TI - Learning--fad or fashion? PMID- 12630302 TI - Do midwives matter? The 12th RCM annual student conference. PMID- 12630303 TI - RCM annual conference 2003: promoting the business of midwifery. PMID- 12630304 TI - The national 'working with teenagers' midwifery network. PMID- 12630305 TI - Case study: Queen Charlotte's hospital. Queen Charlotte's one-to-one 'young mums' midwifery scheme. PMID- 12630306 TI - The RCM virtual institute for birth: promoting normality. PMID- 12630307 TI - Supervision in midwifery practice. PMID- 12630308 TI - What men think of midwives. AB - It is now widely accepted that woman-centred maternity care is important. But surely planners and service providers should also examine the needs of expectant fathers? A postal survey of a randomly selected sample of 837 fathers-to-be throughout the UK found that midwives are not meeting all men's information and support needs. Although midwives were more highly rated than GPs and hospital doctors, men felt that midwives could still listen to them more, enable them to ask questions and explain things to help them better understand physical processes, clinical procedures, the baby's behaviour and their partner's needs. Most men wanted to be involved in their partner's pregnancy and care, but many felt left out by health professionals. Men play a pivotal role in supporting their partner during pregnancy and influence women's baby-feeding choices and esteem after giving birth. It is crucial that midwives see men not as an extra burden, but as individuals with needs of their own who are usually the main supporters of the women and babies at the centre of midwifery care. PMID- 12630309 TI - The Midwifery Interactive Learning Environment (MILE). AB - The aim of the MILE project is to foster a multidisciplinary collaboration between the healthcare sector, specifically healthcare professionals and IT and computing experts. The objective is to develop a computer-based learning (CBL) environment for midwifery and nurse education. As there is little, if any, work within this application domain, MILE will be developed covering an appropriate subset of common skills necessary for midwives and nurses to work within obstetrics and/or gynaecology. The scope and functionality of the CBL environment will enable evaluation of the uptake and performance of such technology within a highly skilled and wide range task environment. This involves both classroom and on-the-job training. PMID- 12630310 TI - Progress in a pocket. PMID- 12630311 TI - Compounds isolated at the Institute of Microbiology in 1989-2001 and future trends. AB - A total of 307 new compounds, natural, semisynthetic or synthetic, were isolated at the Institute of microbiology during the last twelve years. Due to the development of separation (chromatographic) methods and of analytical methods used to determine the chemical structure of these compounds, i.e. NMR, MS and X ray diffraction, many new metabolites could be described. PMID- 12630312 TI - Identification of the EcoKI and EcoR124I Type I restriction--modification enzyme subunits by non-equilibrium pH gradient two-dimensional gel electrophoresis. AB - Effectively optimized and reproducible procedure for monitoring the composition of type I restriction-modification endonucleases EcoKI and EcoR124I by non equilibrium pH gradient two-dimensional (2-D) gel electrophoresis is described. Three subunits of the enzyme complex, which widely differ from one another in their isoelectric points and molar mass, were identified in crude cell extracts of E. coli. For the first time all three subunits of both EcoKI and EcoR124I were detected as distinct spots on a single 2-D gel. A sensitive immunoblotting procedure was suggested suitable for routine use in determining the identity of individual subunits. Potential application of this method for detailed studies of regulation of the function and stoichiometry of the enzyme complexes is discussed. PMID- 12630313 TI - PCR-based methods for identification of Enterococcus species. AB - Two DNA-based techniques were used for species identification of enterococci. PvuII digestion of the genus-specific PCR product yielded four different restriction profiles among 20 enterococcal species; one of them was species specific for E. faecium. In the second case, 32 reference strains belonging to 20 enterococcal species were divided to 12 groups by amplification of internal transcribed spacer of rRNA operon. Interspecies and some intraspecies profile variability was determined. Both methods gave similar results. PMID- 12630315 TI - Effects of nitrogen sources on bacteriocin production by Enterococcus faecium A 2000. AB - The production of a novel broad-spectrum antimicrobial peptide enterococcin A 2000, active against Gram-positive and Gram-negative microorganisms including Listeria subsp. and Escherichia coli, by Enterococcus faecium strain A 2000 isolated from the surface of traditional Bulgarian yellow cheese "kash-kaval" is considerably influenced by complex nitrogen sources in the production medium. Medium components, especially peptone and yeast extract, and their concentration contributed to the increase in bacteriocin production during the stationary phase (16-46 h) of cultivation even in the absence of one of the components present in the basal cultivation MRS medium. PMID- 12630314 TI - Presence and cellular distribution of soybean agglutinin-binding epitopes in two strains of Bacillus subtilis. AB - The cellular location of N-acetylgalactosamine in Bacillus subtilis strains 168 and 170 was examined by electron microscopy using gold-conjugated soybean agglutinin (SBA) as a marker. Post-embedding labeling of sectioned material showed SBA-reactive, galactosamine-containing polymers associated with the cell membrane or the cytoplasm of the two strains. This intracellular location was distinct from the concanavalin A-binding epitopes that were located over the cell wall. Labeling of whole cells (native, fixed in glutaraldehyde, or treated with proteinase K, or Tween 20) before negative staining revealed no galactosamine exposed on the surface of strain 168. On the surface of strain 168 some exposed galactosamine terminal residues were detected; their accessibility to SBA increased when Tween 20 or proteinase K was applied. PMID- 12630317 TI - Toxicological response of the green alga Scenedesmus bijuga to mercury and lead. AB - Effect of mercury or lead on the growth, bioaccumulation and some enzyme activities of one of the most common algae in River Nile, Scenedesmus bijuga, was determined. The cell count and chlorophyll a content decreased with an increase in mercury or lead concentrations in a culture medium, particularly at higher doses. Higher mercury and lead uptake was observed with increasing concentration of the elements. The alga accumulated appreciably more mercury than lead. At higher doses, the two elements strongly suppressed some enzyme activities of primary S. bijuga metabolism. PMID- 12630316 TI - The GCR1 gene function is essential for glycogen and trehalose metabolism in Saccharomyces cerevisiae. AB - Trehalose (Tre) and glycogen (Glg) are synthesized in response to unfavorable growth conditions from glycolytic intermediates in Saccharomyces cerevisiae. Transcription of the glycolytic genes is activated by the Gcr1p complex, the DNA binding transcription factor that directly associates with the CT-box sequences on the promoter region of the glycolytic genes. gcr1 mutant yeast cells cannot utilize glucose effectively. Glg and Tre levels in stationary-phase gcr1 mutant yeast cells were 20-50% of those in the wild-type strain. Likewise, stress induced accumulation of Tre and Glg in gcr1 mutant cells was significantly lower than in the wild type. In addition, both the synthesis and the degradation of Tre and Glg are very slow in the gcr1 mutant. It seems that Gcr1p function is essential for the coordinated regulation of glycolysis, Tre and Glg metabolism in S. cerevisiae. PMID- 12630319 TI - Effect of rice--glycerol complex medium on the production of Lovastatin by Monascus ruber. AB - Response surface methodology (RSM) was employed to study the effect of the composition of the rice-glycerol complex medium on the production of lovastatin (Lvs) by the ascomycete Monascus ruber in mixed solid-liquid (or submerged) cultures at 25 degrees C. Four components (rice powder, peptone, glycerol, glucose) were studied to evaluate the approximate polynomial for all dependent variables, explaining their effects on the production of Lvs. The best composition derived from RSM regression was (in g/L) rice powder 34.4, peptone 10.8,, glucose 129, KNO3 8.0, MgSO4.7H2O 4.0 and glycerol 36.4 mL/L. With this composition, the Lvs production was 157 mg/L after 10 d of cultivation. In comparison with glycerol and glucose, the rice powder becomes a more suitable carbon source and represents a great potential for the production of Lvs. PMID- 12630318 TI - Purine degradation in the edible mushroom Agaricus bisporus. AB - Agaricus bisporus is able to use urate, allantoin, allantoate, urea and alloxanate as nitrogen sources for growth. The presence of urate oxidase, allantoinase, ureidoglycolase and urease activities, both in fruit bodies and mycelia, points to a degradative pathway for urate similar to that found in various microorganisms. So far all efforts to demonstrate the enzyme responsible for allantoate degradation failed. A urease inhibitor appeared to be present in cell-free extracts from fruit bodies. PMID- 12630320 TI - Purification and characterization of two beta-glucosidases from the thermophilic fungus Thermoascus aurantiacus. AB - beta-Glucosidase from the fungus Thermoascus aurantiacus grown on semi-solid fermentation medium (using ground corncob as substrate) was partially purified in 5 steps--ultrafiltration, ethanol precipitation, gel filtration and 2 anion exchange chromatography runs, and characterized. After the first anion exchange chromatography, beta-glucosidase activity was eluted in 3 peaks (Gl-1, Gl-2, Gl 3). Only the Gl-2 and Gl-3 fractions were adsorbed on the gel matrix. Gl-2 and Gl 3 exhibited optimum pH at 4.5 and 4.0, respectively. The temperature optimum of both glucosidases was at 75-80 degrees C. The pH stability of Gl-2 (4.0-9.0) was higher than Gl-3 (5.5-8.5); both enzyme activities showed similar patterns of thermostability. Under conditions of denaturing gel chromatography the molar mass of Gl-2 and Gl-3 was 175 and 157 kDa, respectively. Using 4-nitrophenyl beta-D glucopyranoside as substrate, Km values of 1.17 +/- 0.35 and 1.38 +/- 0.86 mmol/L were determined for Gl-2 and Gl-3, respectively. Both enzymes were inhibited by Ag+ and stimulated by Ca2+. PMID- 12630321 TI - Decolorization of synthetic dyes by Pleurotus ostreatus isolates differing in ligninolytic properties. AB - The ability to decolorize four synthetic dyes (Phenol Red, Evans Blue, Eosin Yellowish and Poly B411) in five Pleurotus ostreatus strains (a parental strain and four isolates derived from it) was determined. Two of the isolates had markedly higher and other two substantially lower production of ligninolytic enzymes and hydrogen peroxide than the parental strain. Like the parental strain, the higher-producing isolates were able to decolorize all the tested dyes, but not to a higher extent than the parental strain. In contrast, two lower-producing isolates exhibited slow decolorization, which was incomplete even at the end of cultivation. Evans Blue and Eosin Yellowish strongly suppressed the growth of the strains, while Phenol Red and Poly B411 induced none or only a very slight growth reduction. PMID- 12630322 TI - Bioremediation of PAH-contaminated soil by composting: a case study. AB - Composting technique was used for bioremediation of industrial soil originating from a former tar-contaminated site. The composting process was regulated by aeration to keep optimal temperature gradient and concentrations of O2 and CO2 inside the composting pile. The efficiency of bioremediation was evaluated by performing analysis of 11 individual three- to six-ring unsubstituted aromatic hydrocarbons (PAH) and estimating of changes in ecotoxicity of the contaminated soil. After 42 d of composting, PAH with 3-4 rings were removed from 42 to 68%, other higher-molar mass PAH from 35 to 57%. Additional 100 d of compost maturation in open-air field did not result in a further decrease of PAH. Ecotoxicity tests performed with bioluminescent bacteria Vibrio fischerii showed a decrease in toxicity both after composting and maturation phases. However, toxicity tests on mustard-seed germination did not reveal any significant changes during composting and maturation phases. PMID- 12630323 TI - Kinetics of phenol oxidation by Candida tropicalis: effects of oxygen supply rate and nutrients on phenol inhibition. AB - The kinetics of phenol degradation was estimated in a fed-batch reactor system. Effects of oxygen and nutrient excess or limitation as well as the presence of several essential ions on the phenol- and oxygen-specific uptake rates achieved simultaneously in a bioreactor were shown. Candida tropicalis was grown on phenol as the only carbon and energy source. Applying the best fit of polynomial function, the maximum specific uptake rates of phenol and oxygen, the critical concentrations of phenol, the half-saturation constants and inhibition constants were determined. Linear relationship between specific phenol uptake rate and the exogenous respiration rate was found regardless of the kind and presence of essential nutrients. At oxygen limitation both the phenol uptake rate and the cell affinity to phenol decreased more strongly compared with those under nutrient limitation. Oxygen in excess resulted in a significant increase of cell tolerance toward phenol. The presence of essential nutrients increased the specific phenol degradation rate and led to complete phenol oxidation. PMID- 12630324 TI - In vitro effects of cAMP-elevating agents and glucocorticoid either alone or in combination on the production of nitric oxide, interleukin-12 and interleukin-10 in IFN-gamma- and LPS-activated mouse peritoneal macrophages. AB - The effects of cAMP-elevating agents, N6-2'-O-dibutyryl cAMP (Bu2cAMP), and glucocorticoid (dexamethasone) on the production of inflammatory mediators- nitric oxide and interleukin-12 (IL-12) and anti-inflammatory mediator interleukin-10 (IL-10) were demonstrated in murine peritoneal macrophages. Inducible nitric oxide synthase (iNOS) and iNOS mRNA were detected by northern blot and western blot, respectively. The cAMP elevating agents Bu2cAMP and prostaglandin E2 each alone did not show any effect on NO production but along with IFN-gamma and lipolysaccharide (LPS) they slightly enhanced NO production. Dexamethasone inhibited NO production in IFN-gamma- and LPS-treated cells; cAMP elevating agents interfered with the NO production inhibited by dexamethasone. Inhibition was revealed at the mRNA level as well as at protein level. Bu2cAMP or dexamethasone either alone or synergistically inhibited IL-12 production; Bu2cAMP interfered with dexamethasone-mediated inhibition of IL-10 production in IFN gamma- and LPS-treated macrophages. The use of glucocorticoids along with cAMP elevating agents was beneficial in lowering the level of inflammatory mediator IL 12 and producing high levels of the anti-inflammatory mediator IL-10 active in cell protection. On the other hand, interference of Bu2cAMP with dexamethasone mediated NO inhibition may have adverse effect. Therefore, adverse effects due to cAMP-mediated interference (inhibition) with NO synthesis may occur in many inflammatory diseases during combined drug therapy by glucocorticoids and cAMP elevating agents. PMID- 12630325 TI - T helper, cytotoxic T lymphocyte, NK cell and NK-T cell subpopulations in patients with chronic hepatitis C. AB - The phenotype of intrahepatic (IHL) and peripheral blood lymphocytes (PBL) was determined, and the production of cytokines by T lymphocytes analyzed in patients with chronic hepatitis C (CHC). Three-color fluorescence-activated cytometric analysis was done for 36 patients with untreated CHC. The percentage of peripheral blood memory T cells was higher in patients with CHC than in healthy controls (all data in %, significant at p < 0.001; 74.6 +/- 2.7 vs. 58.3 +/- 4.5), and a greater proportion of them were observed in the intrahepatic compartment (IHL-94.2 +/- 2.8 vs. PBL-74.6 +/- 2.7). There was a higher percentage of peripheral blood T helper 1 lymphocytes expressing IFN-gamma (IFN gamma/IL-4) in these patients (4.6 +/- 0.7 vs. control-2.2 +/- 0.5). The expression of CXCR3 chemokine receptors on peripheral blood T helper cells was also high compared with the control (39.8 +/- 4.8 vs. 26.8 +/- 2.5) and a large percentage of T cells expressing CXCR3 or CCR5 chemokine receptors was observed in hepatitis C virus (HCV)-infected liver (CXCR3: IHL vs. PBL-74.9 +/- 5.7 vs. 39.8 +/- 4.8; CCR5: IHL vs. PBL-65.9 +/- 5.9 vs. 19.1 +/- 2.1). The intrahepatic compartment contains a greater proportion of activated cytotoxic T lymphocytes (CTL) and natural killer-T (NK-T) cells than peripheral blood (CTL: IHL vs. PBL 69.5 +/- 3.2 vs. 59.9 +/- 3.1; NK-T: IHL vs. PBL-10.6 +/- 2.5 vs. PBL: 3.99 +/- 0.5). The data suggest that in HCV-infected subjects, memory TH1 lymphocytes, activated CTL and NK-T cells compartmentalize in liver tissue and could play an important role in pathogenesis of chronic hepatitis. PMID- 12630326 TI - Detection of pap-, sfa- and afa-specific DNA sequences in Escherichia coli strains isolated from extraintestinal material. AB - P-fimbriae, S-fimbriae and AFA-adhesins are virulence factors responsible for adherence of Escherichia coli strains to extraintestinal host-cell surface. Detection of pap-, sfa- and afa-specific sequences performed by PCR revealed 74% pap+, 65% sfa+, and 8.3% afa+ strains in a group of 84 extraintestinal E. coli isolates. Detection in a group of fecal strains showed 29% pap+, 21% sfa+ and 4% afa+ strains. pap together with sfa were found as the most frequent combination (56%) among extraintestinal isolates probably due to localization of pap- and sfa operons on a common pathogenicity island. The occurrence of afa-specific sequence among 56 urine strains was 11%, although no afa+ strain was detected among 28 gynecological isolates. No strains with detected adhesin operons were found among twenty (24%) extraintestinal E. coli strains. PMID- 12630328 TI - Detection of anti-Candida antibodies by the indirect immunofluorescence assay in patients with cancer in the orofacial region. AB - An indirect immunofluorescence assay was performed to detect antibodies to Candida albicans blastospores and germ tubes. Serum specimens were obtained from 82 patients with neoplastic diseases in the orofacial region and thrush of the oral mucosa. C. albicans was identified in the oral cavity of 63 patients investigated but serum anti-Candida antibodies were detected in only 23 of them. Serological examination showed that titers of antibodies to C. albicans blastospores ranged from 1:20 to 1:1280. High titers from 1:640 to 1:1280 were detected in patients without antibiotic, cytostatic, or radiotherapeutic treatment. The titers of antibodies to C. albicans germ tubes ranged from 1:20 to 1:640. Our results indicate that titers of antibodies to the C. albicans germ tubes were lower and were detected in a smaller number of patients. PMID- 12630327 TI - The frequency of Candida species in onychomycosis. AB - Mycological investigation of 108 nail specimens taken from a total of 41 patients examined over three years included direct microscopy and repeated cultures. A higher incidence of onychomycosis of the fingernails (75%) was observed in women while afflictions of the toenails (71%) prevailed in men. The highest prevalence of onychomycosis was found in patients between 50 and 70 years of age. Candida albicans was the dominant organism causing onychomycosis (prevalence rate 60.9%), followed by C. parapsilosis (19.6%), C. tropicalis (9.8), C. krusei (4.9), C. guilliermondii and C. zeylanoides (2.4% each). PMID- 12630329 TI - Infective endocarditis caused by unusual gram-positive pathogens. AB - Of a total of 81 patients hospitalized in the infectious diseases department in 1990-2000 with infectious endocarditis caused by Gram-positive pathogen, unusual etiological agents were found in several cases: Streptococcus pyogenes, Streptococcus pneumoniae, Corynebacterium diphtheriae, and Gemella morbillorum. Cardiac defects were present in the latter two patients: bicuspid aortic valve and tetralogy of Fallot. Two patients were successfully treated with antibiotics only and one patient with antibiotics and surgery. The patient with C. diphtheriae endocarditis died due to progressive sepsis and multiple organ failure. PMID- 12630330 TI - Fatty acid analysis of Stenotrophomonas maltophilia clinical strains showing different susceptibility to antibiotics at 30 and 37 degrees C. AB - Isolates of Stenotrophomonas maltophilia species display the feature "temperature dependent susceptibility" (TDS) to antibiotics. Both 30TDS strains (at least 4 times lower value of minimum inhibitory concentration (MIC) of an antibiotic at 30 than at 37 degrees C) and 37TDS strains (at least 4 times lower value of MIC at 37 than at 30 degrees C) were described. Changes in the distribution of saturated and unsaturated fatty acids (FA) at 30 and 37 degrees C were considered as one of possible causes of the TDS phenomenon. Gas chromatography was used to determine the distribution of individual FA in five 37TDS strains of S. maltophilia (Group I); in five strains with MIC values unaffected by the cultivation temperature (Group II) and in six 30TDS (four strains) or 30/37TDS (two strains) isolates (Group III). At identical temperatures, no statistically significant differences in the distribution of major FA (iso-15:0, anteiso-15:0, 16:0 and 16:1) were registered between individual groups. Statistically significant (p < 0.05) differences between groups were found in minor FA only (iso-16:0, iso-17:0 and iso-17:1). Distribution changes of cellular FA at 30 and 37 degrees C can be considered to play only a minor role in the formation of the TDS phenomenon. PMID- 12630331 TI - Production of interferons and change of the lymphocyte subpopulation phenotype in peripheral blood at cervical papillomavirus infection. AB - IFN-gamma and IFN-alpha production in vitro by peripheral blood cells activated by phytohemagglutinin or the Newcastle disease virus was impaired in patients with a benign process, cervical intraepithelial neoplasm and cancer in situ associated with human papillomavirus infection. In case of IFN-gamma and IFN alpha production impairment following cervical papillomavirus infection, the increased severity of disease was accompanied by remarkable IFN system suppression. The lower synthesis of both IFN correlated with changes of some lymphocyte-subpopulation phenotype in peripheral blood. Lower CD4+ and CD3+ DR+ T cell concentrations were observed in papillomavirus-infected patients with impaired IFN production; impaired IFN-gamma production was accompanied by lower CD4/CD8 index. PMID- 12630332 TI - The serologic screening for celiac disease in the general population (blood donors) and in some high-risk groups of adults (patients with autoimmune diseases, osteoporosis and infertility) in the Czech republic. AB - The prevalence of celiac disease (CD) was determined in healthy blood donors and in high-risk groups of adults (a total of 1835 adults--randomly selected 1312 healthy blood donors, 102 patients with primary osteoporosis, 58 patients with autoimmune diseases and 365 infertile women). It was calculated on the basis of a two-step serologic screening method--in the first step IgA and IgG antigliadin antibodies (AGA) and IgA anti-gamma-glutamyltransferase ('transglutaminase') antibodies (ATG) were estimated, in the second step sera positive for IgA AGA and/or IgA ATG were examined for antiendomysial IgA (AEA) antibodies. Immunoenzymic assay (ELISA) was used for determining of AGA and ATG antibodies; immunofluorescence method, performed on human umbilical cord tissue, was used for assaying of AEA antibodies. Total serum IgA level in only IgG AGA positive subjects was measured by routine turbidimetric method. 0.45% of healthy blood donors, 0.98% of osteoporotic patients, 2.7% of patients suffering from autoimmune disease and 1.13% of women with infertility considered as immunologically mediated were found to be positive in both steps of serologic screening (AGA and/or ATG and antiendomysium positive). The presumed high prevalence of seropositivity for CD in apparently healthy Czech adult population was confirmed. In the high-risk groups, the prevalence of seropositivity for CD was approximately 2-4 times higher than in healthy blood donors. The real prevalence of CD in the tested groups, however, can be estimated after performing small intestinal biopsy in the seropositive patients. PMID- 12630334 TI - Using oral amiodarone safely. AB - Amiodarone is licensed for the treatment of severe cardiac rhythm disorders where other treatments either cannot be used or have failed. Because of safety concerns, the summary of product characteristics (SPC) recommends that amiodarone "should be initiated and normally monitored only under hospital or specialist supervision". However, the responsibility for prescribing and monitoring maintenance therapy often falls to non-specialists. Here, we discuss safety issues relating to the long-term use of oral amiodarone in adults. PMID- 12630333 TI - Differential effect of Bacillus firmus on immune response and enterocyte brush border enzyme levels in BALB/c and B10.BR mice. AB - A nonpathogenic bacterium of external environment possessing remarkable immunomodulatory activity, Bacillus firmus (BF) inactivated with formaldehyde, was given intragastrically to two genetically different mouse strains BALB/c (H 2d) and B10.BR/SnPh (B10.BR, H-2k) reared in conventional (CV) and B10.BR strain also in germ-free (GF) conditions. Repeated intragastric administration of BF (500 micrograms every other day over two weeks, starting at the age of 3 months) significantly enhanced intestinal IgA levels in CV BALB/c mice but did not affect intestinal IgA in CV B10.BR mice. In GF B10.BR mice, IgG levels in sera and intestinal washings increased after BF administration compared to CV B10.BR mice. In CV BALB/c mice, specific activity of enterocyte brush-border enzymes (lactase, gamma-glutamyltransferase, alkaline phosphatase) decreased after BF treatment; sucrase (sucrose alpha-glucosidase) activity was not affected. On the other hand, in B10.BR mice, specific activity of gamma-glutamyltransferase and dipeptidyl peptidase IV were higher after administration of BF in both CV and GF groups relative to untreated controls. The activities of lactase and glucoamylase (glucan 1,4-alpha-glucosidase) were significantly stimulated only in the group of GF B10.BR mice treated with formolized BF. The stimulation of immunoglobulin production after BF treatment was accompanied by changes in the levels of enterocyte brush-border enzymes; this responsiveness to BF treatment was genetically regulated. PMID- 12630335 TI - New topical drugs for open-angle glaucoma. AB - When we reviewed the management of open-angle glaucoma in 1997, the topical drug treatments available included beta-blockers, a carbonic anhydrase inhibitor (dorzolamide), an alpha 2 agonist (apraclonidine), a prostaglandin analogue (latanoprost) and a variety of miotics and sympathomimetics. We concluded that a beta-blocker was the treatment of first choice. Here, we review seven new topical preparations that have been marketed, or had their licensed indications changed, since our earlier article. PMID- 12630336 TI - Tiotropium for chronic obstructive pulmonary disease. AB - In 2001, we concluded that patients with symptoms of stable chronic obstructive pulmonary disease (COPD) inadequately controlled by an 'as-required' inhaled short-acting beta 2 agonist can be helped by regular use of an inhaled antimuscarinic bronchodilator or a long-acting beta 2 agonist. Since then, a new antimuscarinic drug [symbol: see text] tiotropium bromide (Spiriva- Pfizer/Boehringer Ingelheim) has been licensed as a once-daily bronchodilator for maintenance treatment of COPD in people aged 18 years or over. Here, we review the evidence for efficacy of tiotropium and discuss its role in the maintenance treatment of COPD. PMID- 12630337 TI - [Relationship between science, politics, religion and daily life]. AB - The relationship between science, politics and religion is discussed, with special reference to the effect of scientific discoveries on the improvement of the quality of everyday life. It is concluded that the results of scientific research lead to prosperity of man and nations. However, the society appears to insufficiently use these advantages which can be partly the result of failing to recognize the connection between basic science and products that re-define everyday life. On the other hand, problems might originate from the aversion towards the risks as well as from short-term planning. PMID- 12630338 TI - [Incidence and distribution of Streptococcus pyogenes type M in patients treated at the Dr. Fran Mihaljevic Infectious Disease Clinic in Zagreb from 1990 to 1996]. AB - AIM: The aim of this study was to estimate the changes in the appearance and distribution of M types of Streptococcus pyogenes in different cultures from 78 patients treated during the 1990-1996 period at the Dr. Fran Mihaljevic University Hospital for Infectious Diseases in Zagreb. MATERIALS AND METHODS: Isolates were characterized by the T-agglutination pattern and M type and/or opacity factor type using the standards recommended by two World Health Organization Collaborating Centers for Reference and Research on streptococci from Minneapolis and Prague. RESULTS: In this study, 19% (15/79) of isolates were recovered from normally sterile sites, 26.6% (21/79) came from skin and 54.4% (43/79) from throat swabs. In one patient isolates from the skin and blood culture were analyzed. Of all, 92.4% (73/79) of the isolates were typed by T agglutination pattern and 73.5% (58/79) by M protein and/or OF typing. The results of M typing showed 14 M types: M1, M3, M4, M5, M6, M11, M12, M28, M57, M58, M60, M75, M76 and M78. The most commonly isolated types were M1 and M3 (13.8%, 8/58 each), followed by M28 found in 12.1% (7/58), and M6 and M12 in 10.3% (6/58) each. These five M types accounted for 60.3% (35/58) of all isolates. Analysis to changes in the distribution of M1 and M3 types during the 1990-1991 and 1992-1993 periods revealed a significantly greater proportion of M1 and M3 isolates in the former (Fisher's two-tailed exact test, p = 0.018). A significantly greater proportion of M1 and M3 isolates was also recorded in the 1990-1991, than in 1994-1996 period. (Fisher's two-tailed exact test, P = 0.021). It was investigated whether Streptococcus pyogenes M1 and M3 types were associated with toxic and invasive infection. There were 28.2% (22/78) of patients with toxic and invasive infection: 31.9% (7/22) of them with the diagnosis of scarlet fever, whereas 68.1% (15/22) of the strains were obtained from normally sterile sites. There were 45.5% (10/22) of M1 and M3 types from patients with toxic and invasive infections. Types M6, M28 and M76 were found in an equal proportion of 9.1% (2/22), and M4, M12 and M 60 of 4.5% (1/22) each. In three strains, M type could not be identified (T8/25 SOR+, T25 SOR+ and T11 SOR+). M1 and M3 types were isolated from 10.3% (6/56) of patients with other streptococcal infections. A significantly greater proportion of M1 and M3 types was recorded in patients with toxic and invasive infections than in those with other streptococcal infections (Fisher's two-tailed exact test, p = 0.004). A multivariate logistic regression model was used to assess whether the increased proportion of streptococcal M types 1 and 3 was associated with the 1990-1991 period, or with the infection characteristics. CONCLUSIONS: The changes in the distribution of M types 1 and 3 were found to be significantly associated only with toxic and invasive infections (odds ratio 4.35, p = 0.025). Odds ratio suggests that patients with toxic and invasive infections had a 4.35--fold increased risk of infections caused by types M1 and M3 found in patients with other streptococcal infections. The increased proportion of M types 1 and 3 during the 1990-1991 period was more significantly associated with the characteristics of infection than with the study period. PMID- 12630339 TI - [Evaluation of the efficacy and safety of lodoxamide in patients with allergic eye diseases]. AB - PURPOSE: To evaluate the effectiveness of lodoxamide in the therapy and prophylaxis of the ocular allergies. PATIENTS AND METHODS: This prospective study included 64 patients divided in 2 groups. In the first group there were 47 symptomatic patients suffering from seasonal allergic conjunctivitis (n = 27), perennial allergic conjunctivitis (n = 16) and giant papillary conjunctivitis (n = 4). The symptomatic patients were examined upon arrival and every 2 weeks until the symptoms were reduced. During the period, lodoxamide drops were administered 4 times a day. In the second group there were 17 patients who had a history of seasonal ocular allergies during previous years but were still not symptomatic upon arrival. In the group of 17 patients who were very likely to develop ocular allergy but up to inclusion into the study had no ocular symptoms, 12 (70%) presented with allergic rhinitis while 5 (30%) had asthma. The symptoms intensity (itching, discomfort, foreign body sensation, pain, tearing) was graded on a 0-3 scale. The clinical signs (follicles, papillae, hyperemia, conjunctival edema, Trantas dots, stromal infiltrates) were detected on a slit lamp examination and graded on a 0-3 scale. RESULTS: In the group of 47 symptomatic patients 2-4 weeks after lodoxamide administration, 70-80% of symptomatic patients examined clinically had less or no follicles, 60-70% had reduced or no edema, the secretion was stopped or greatly reduced in 50-70% of patients. All of the patients reported reduced or no itching, 60% of patients reported less discomfort, photophobia and reduced tearing. In the asymptomatic group two weeks after the initiation of the prophylaxis 88% of patients were still with no ocular signs of allergy while 12% of patients had a conjunctival hyperemia and follicles. After 4 weeks of the prophylaxis 76% of patients patients were still asympthomatic while only 4 developed a mild form of allergic conjunctivitis (conjunctival hyperemia and papillae), but less severe than during previous episodes. CONCLUSION: Lodoxamide was effective in reduction of symptoms and clinical signs of the ocular allergies. The best results were obtained if the drug was administered as a prophylaxis or very early in the course of the disease. PMID- 12630340 TI - [Evaluation of vascular risk factors in primary open-angle glaucoma using doppler sonography]. AB - Primary open angle glaucoma (POAG) is a syndrome of progressive optic neuropathy characterized by optic nerve head excavation and visual field defects. Poor correlation between intraocular pressure (IOP) and progression of glaucoma disease puts vascular mechanism in the center of attention. By color Doppler sonography, quantification of blood flow changes in the vessels which supplying optic nerve head is possible. AIM: To examine the antiglaucomatous action of selective-blockers by color Doppler sonography. PATIENTS AND METHODS: Forty-four patients (88 eyes) were divided into two groups: group 1 treated with selective beta-blockers and group 2 treated with nonselective beta-blockers. Vascular indices (RI, PI) were measured in the central retinal artery and posterior ciliary arteries. RESULTS: Decreased blood flow and increased vascular indices were found in both groups, with a statistically significant difference between group 1 and group 2. Blood flow velocity was higher and vascular indices were lower in group 1 (Betoptic 0.5%) than in group 2 (Timolol 0.5%). CONCLUSIONS: beta-blockers, selective (betaxolol-calcium channel blockers) have more potent vasoactive and neuroprotective action. Color Doppler sonography is useful in the quantification of blood flow changes in the vessels in patients with POAG. PMID- 12630341 TI - [Neurologic sequelae of bone changes in multiple myeloma and its therapy]. AB - Multiple myeloma (MM) is a plasma cell malignancy characterized by infiltration of bone marrow, bone destruction, infiltration of soft tissues with plasma cells, and suppression of normal hematopoiesis. The production of monoclonal immunoglobulins with or without light chains is a major feature of the disease. Full spectrum of plasma cell dyscrasias include monoclonal gammapathy of undetermined significance, smouldering myeloma, indolent multiple myeloma, and fully developed, symptomatic multiple myeloma. The usual presenting features of MM include bone pain, weakness, fatigue, fever and infection. Neurologic symptoms are less common but one must not forget that MM may present with a neurologic disease. Careful neurologic history and examination are mandatory in patients with MM. Neurologic symptoms may be a direct manifestation of MM or may be due to the immune effect of monoclonal proteins directed against different neural structures. Finally, metabolic consequences (uremia, hypercalcemia, hyperviscosity) of MM may produce a broad spectrum of different neurologic symptoms including headache, blurring of vision, drowsiness, precoma, coma, vertigo, ataxia, hemiparesis and epileptiform seizures. The most common location of bone changes in MM is the thoracic spine, where it causes osteolytic changes with consequent compressive fractures. The most disastrous sequel is paraplegia. Multiple vertebral involvement with the evidence of osteolytic changes in other bones is usual, but solitary vertebral myeloma may occur. Myeloma usually involves the bone of the vertebral body and then spreads into the extradural space. However, patients with solitary extradural myeloma have been reported. Skull myeloma is frequently asymptomatic. It may grow externally or, rarely, there is intracranial expansion. Involvement of the cranial nerves is not rare, with II, V, VI, VII and VIII cranial nerves being most often affected. Isolated intracerebral plasmacytomas are extremely rare. Diagnostic approach includes plain X-rays of the skeleton, which was found to be the method of choice for demonstration of osteolytic changes, whereas magnetic resonance with gadolinium enhancement most reliably displays the degree of vertebral involvement and demonstrates any associated soft tissue mass. Current treatment of osteolytic changes in multiple myeloma include chemotherapy, radiotherapy in combination with dexamethasone, monthly infusions of bisphosphonates, surgical decompression, and kyphoplasty. Therapeutic approach is dictated by the presenting symptoms. In case of pain as the predominant symptom, treatment with chemotherapy and radiotherapy may be appropriate. Compressive symptoms are relieved with dexamethasone followed by radiotherapy and chemotherapy. Surgical decompression is used in patients with vertebral collapse and vertebral instability. Kyphoplasty is a new method used in the treatment of osteolytic changes of vertebral bodies. A viscous cement is injected into the cavity by a balloon-like inflatable bone tampon. It has been successfully employed to improve the quality of life, to reduce pain, and to increase overall functioning in patients with vertebral compression fractures by restoring most of the original height of the vertebral body. Bisphosphonates reduce pain associated with osteolytic changes in multiple myeloma, but also significantly reduce skeletal events (pathologic fracture, spinal cord compression, surgery or irradiation of bone) via unknown mechanism. It seems that bisphosphonates, by inhibiting bone resorption, alter the microenvironment in which the MM cells grow. PMID- 12630342 TI - [Intrauterine hypoxia and sudden infant death syndrome]. AB - Sudden infant death syndrome (SIDS) or crib or cot death are synonyms for the sudden, unexpected and unexplained death of an infant. The incidence of SIDS has been estimated to be from 1-2% to 3%. Protracted intrauterine hypoxia or recurrent hypoxic insults during fetal life undoubtedly influence the development of the central nervous structures as a tissue most susceptible to hypoxia, although well developed mechanisms of defense against hypoxia exist during the fetal life. The mechanisms underlying SIDS include neurologically compromised infants who are deprived of compensatory mechanisms during sleep, sustaining a hypoxic insult with alterations in neurotransmitter receptors within the regions involved in chemoreception and cardiovascular control. Changes in the brain result from perinatal prolonged hypoxia (persistent reticular pathways in the pons and medulla, astroglia in the brainstem, gliosis of brain nerve nuclei, defects in neurotransmitter receptors, neuronal apoptosis, microthrombosis, and hypoxic ischemic lesion). Hypoxic perinatal risk factors for SIDS included passive and active exposure to cigarette smoking in pregnancy, abuse of drugs, alcohol, coffee and medication in pregnancy, intrauterine growth retardation, perinatal hypoxia with or without resuscitation, preeclampsia, anemia in pregnancy, prematurity, multiparity, multiple pregnancy, pregnant women aged < 20 years and > 35 years, cardiocirculatory, pulmonary and endocrine diseases in pregnancy, and short time interval between two pregnancies. As cigarette smoking has been demonstrated to lead to fetoplacental insufficiency, which result in fetal hypoxia, it is concluded that hypoxia is a precondition for the occurrence of SIDS. Prenatal exposure to cigarette smoke decreases maternal red blood cell count, and concentrations of tyrosine and selenium, reduces fetal and neonatal cerebral blood flow, and increases maternal MCV, leukocytosis, especially neutrophils, monocytes and lymphocytes, maternal and fetal heart rate, systolic and diastolic blood pressure, resistance index in umbilical artery, fetal hemoglobin, cytokine, serotonine, dopamine, catecholamine, hypoxanthine, endorphin and interleukin-6. Pregnancy at a risk of hypoxia, especially in heavy smokers, is a major risk factor for SIDS, and such pregnancy requires close and intensive antenatal monitoring. PMID- 12630343 TI - [Mesenteric cysts]. AB - INTRODUCTION: Mesenteric cysts and cystic mesenteric tumors are very rare abdominal growths. They may be localized all over the mesentery, from duodenum to rectum, however, they are mostly found in the ileum and right colon mesentery. There are several classifications of these formations, among which the one based on histopathologic features including 6 groups has been most commonly used: 1) cysts of lymphatic origin--lymphatic (hilar cysts) and lymphangiomas; 2) cysts of mesothelial origin--benign or malignant mesothelial cysts; 3) enteric cysts; 4) cysts of urogenital origin; 5) dermoid cysts; and 6) pseudocysts--infectious or traumatic etiology. PATIENTS AND METHODS: Two adult female patients treated at the Department of Surgery, Zabok General Hospital, are presented. The diagnosis of mesenteric cyst was based on explorative laparotomy indicated for a cystic abdominal growth and characteristic palpatory finding, US and CT findings. In both patients, the cysts were successfully treated by total cystectomy. Pathohistologic findings pointed to lymphatic cysts. Control US finding at 3 months postoperatively was normal in both patients. DISCUSSION: Cystic lymphangioma mostly occurs in the first decade of life, with a female predominance. It is usually accompanied by acute abdominal symptomatology. Lymphatic cysts occur later in life (1:100,000 in adults and 1:20,000 in children), also show female predominance, and as a rule are asymptomatic. A mesenteric cyst, especially lymphatic, should be suspected in the presence of painless abdominal tumor, with occasionally painful abdominal pressure, normal laboratory findings, and good general condition in a female patient. In symptomatic cases, acute or chronic abdominal pain is the most common feature, whereas other symptomatology depends on the localization, size and consequential abdominal organ compression (intestinal obstruction, hydronephrosis, lower extremity lymphedema). The term of cystic mesenteric tumor is mostly used to refer to cystic lymphangiomas and lymphatic cysts. In the former, smooth muscle tissue is found, with endothelial lining towards the cavity. The wall of hilar mesenteric cysts does not contain smooth muscle tissue, however, they also show endothelial lining towards the cavity. Exact differentiation between these two entities is necessary for the disease prognosis. Lymphangiomas are prone to recurrence and infiltrating growth. The diagnosis should be made by use of all standard methods of abdominal tumor diagnosis, with ultrasonography (US) and computed tomography (CT), and especially nuclear magnetic resonance providing most information of the growth size and localization. Total cystectomy is the therapeutic method of choice. Open method has been preferred, although reports on successful cystectomy by the laparoscopic method have already appeared in the literature. CONCLUSION: Intraoperative differentiation between lymphatic cyst and lymphangioma is of utmost importance, and can only be achieved by pathohistologic examination of the cyst wall. If intraoperative biopsy cannot be performed or the finding is uncertain, each cyst should be extirpated in toto due to the above mentioned risk associated with cystic lymphangioma. PMID- 12630344 TI - [Evaluation of the usefulness of three collections of bacteriophages for typing methicillin-resistant Staphylococcus aureus, isolated in Moscow hospitals]. AB - The typing of S. aureus methicillin-resistant strains, isolated in different hospitals of Moscow; was carried out with the use of three collections of phages: the International Set of Phages; the set of phages of the International Center of S. aureus phage typing in London (L); and the experimental collection of phages of the Gamaleya Institute of Epidemiology and Microbiology in Moscow (M). In this study made with the use of both the phages of the International Diagnostic Set and phages L in the standard typing dose of 1 TP about 6% of the cultures under study proved to be sensitive. When the typing dose was increased to 100 TP the phages of the international diagnostic set lyzed 75.5% of the cultures. The typed strains were found to belong to phage types 77 (71.7%), 77/84/85 (19.6%) and 94/96 (6.5%). At a concentration of 100 TP phages L lyzed 83.7% of the cultures, but the dominating phage types could not be determined due to a great variety of phage markers. In contrast to the two preceding collections, the third phage collection M was composed in such a way that in the study of the investigated culture the specificity of its restriction modification was primarily evaluated and only then the presence of antiphage immunity was determined. This latter collection was used in the evaluation of 93.1% of the cultures. By the specificity of their restriction specification system the majority of them were classified with two new groups, heretofore not described. Only this collection M made it possible to differentiate epidemic and sporadic strains and to evaluate the epidemic situation in all 6 hospitals. PMID- 12630345 TI - [Bacteriological monitoring of infective agents of purulent septic diseases in an emergency hospital]. AB - During the period of 1995-2001 bacteriological examination of 13,842 patients with purulent septic diseases (PSD) was carried out. The statistical processing of data revealed that equalization of the dynamic rows of intensive and extensive values characterizing the number of patients with PSD caused by etiologically important infective agents made it possible to evaluate the reliability of information. A trend to increased etiological importance of the genera Pseudomonas, Escherichia and Klebsiella was established. The number of diseases caused by Staphylococcus, Streptococcus, Enterococcus and Proteus decreased. PMID- 12630346 TI - [Biliary microflora of patients with chronic pancreatitis]. AB - Data on the contamination of bile specimens (1997 and 2001, 205 specimens, 137 cultures), examined in the Laboratory of Clinical Microbiology of the Vladimirsky Regional Research Clinical Institute, as well as data on the microflora composition in 40 specimens of bile, obtained from patients with clinically diagnosed chronic pancreatitis at the stage of exacerbation, are presented. Out of 40 specimens of bile 54 cultures of microorganisms were isolated; of these, 33.3% were bacteria of the genus Staphylococcus, Gram negative bacilli took the second place in occurrence (27.8%) and the third place belonged to bacteria of the genus Streptococcus (18.5%). The antibiotic sensitivity of the isolated microorganisms is characterized. PMID- 12630347 TI - [Functioning of opisthorchiasis-tuberculosis parasitocenosis at different stages of invasion (experimental investigation)]. AB - In experiments on golden hamsters the mutual influence of the co-members of opisthorchiasis-tuberculosis parasitocenosis has been established. This mutual influence depends on the stage of invasion and the state of the immune reactivity of the host. After additional immunization of the experimental animals with sheep red blood cells (SRBC) parasitocenosis formed special immune response different from those observed in both opisthorchiasis and tuberculosis. In cases of mixed pathology (opisthorchiasis-tuberculosis), reproduced at the acute stage of invasion, additional body reserves are seemingly switched on, thus leading the immunity system to increased functioning with respect to specific and heterologous antigens. In the chronization of invasion a decreased formation of antibodies to heterologous antigens (SRBC) together with a simultaneous decreased level of specific antiopisthorchiasis and antituberculosis immune response suggest that the reserve capacities of the host immune system, especially of humoral factors, are exhausted. PMID- 12630348 TI - [Epidemiological characterisation of acute and chronic hepatitis B in Uzbekistan]. AB - The epidemiological survey of 126 foci with patients having acute hepatitis B (AHB) and 120 foci with patients having chronic hepatitis B (CHB) was conducted. The observation of the susceptible members of the family showed that a significantly higher level of infection was found in persons having contacts with CHB patients (44.4 +/- 2.3%) in comparison with the members of the families of AHB patients (33.2 +/- 2.3%). The study revealed that children under 14 years were actively involved into the epidemic process; in these children the highest levels of infection were observed in the families of AHB patients (40.2 +/- 3.7%) and CHB patients (57.1 +/- 3.5%). High detection rate of HbsAg were noted in brothers and sisters in the foci of AHB (42.3 +/- 6.4%) and the foci of CHB (52.3 +/- 5.4%), also in parents: 32.4 +/- 5.2% and 46.5 +/- 4.2%, in children: 28.8 +/ 3.4% and 35.6 +/- 3.6% respectively. PMID- 12630349 TI - [Liver cirrhosis and liver cancer associated with viral hepatitis B and C in republic of Tatarstan]. AB - Clinical and epidemiological data on 93 patients with liver cirrhosis, treated in the Gastroenterological Department of the Republican Clinical Hospital of the of the Republic of Tatarstan MoH in 1998-2000 were analyzed. The retrospective analysis of 856 cases of the liver cancer, diagnosed in the Republic of Tatarstan in 1996-2000, was made. In 20.4% of patients with liver cirrhosis and 16.2% of patients with liver cancer, registered in the Republic of Tatarstan, the serological markers of hepatitis B (HB) and hepatitis C (HC) were detected. The domination of HC markers was observed in the liver cirrhosis and that of HB markers in the cancer of the liver. The liver cirrhosis with HC markers was more often registered in the age group of 45-60 years and with HB markers at the age over 60 years, while the cancer of the liver was more often registered in persons aged 75 years and older. The necessity of introducing the official registration of the liver cirrhosis and cancer associated with chronic infection caused by hepatoviruses B and C is confirmed. PMID- 12630350 TI - [Molecular biological monitoring of the polioviruses circulation in Belarus]. AB - A total of 135 polioviruses (PV), including 25 wild and 110 vaccine-related, isolated in Belarus in 1957-1999 were studied by the analysis of the polymorphism of the restriction fragments lengths of two distal regions of the genome: the region (480 oligonucleotide pairs) coding the N-terminal fragment of capsid protein VP1 (RLFP-1) and the region (291 oligonucleotide pairs) coding the N terminal fragment of nonstructural protein of 3D-polymerase (RLFP-3D1). The genetic analysis of the viruses made it possible to determine 3 epidemiologically different periods of PV circulation: (1) the prevaccination period (1957-1959) when wild PV of all 3 serotypes circulated on the territory of Belarus; (2) the early period of the use of Oral Poliomielytis Vaccine (1960-1966), characterized by simultaneous circulation of wild and vaccine PV, as well as vaccine/wild recombinant PV; (3) the period of the elimination of wild PV of indigenous origin and the circulation of vaccine-related viruses (1967-1999). The characteristic feature of wild PV was their pronounced genetic variability. 8 genetic variants of PV1, including 4 genetic groups, 2 genetic variants of PV2 and 1 genetic variant of PV3 were detected; 2 vaccine/wild recombinant PV were detected in 1960 and 1966. More than 40% of the vaccine-related PV under study had altered genetic characteristics (mutations and/or recombinations. Reverse variability, linked with the loss of a number of signs of attenuation, was shown to be characteristic of vaccine PV1. Recombinants occurred most frequently among PV3 (44.9%) and PV2 (40.0%), their recombinations being formed mainly with PV1. Recombinants PV2/PV1 and PV3/PV1 were found to have high frequency of reversion in the "PV1" fragment of the genome; this frequency exceeded that in PV1 with the homotypical genome (66.7 and 44.4% in contrast to 12.5%). PMID- 12630351 TI - [Multicomponent vaccine VP-4 in the therapy of allergic diseases]. AB - In the present study the results of the polycomponent vaccine B[symbol: see text] 4 use for the therapy of patients with bronchial asthma (BA) and latex allergy were generalized. The vaccine was introduced by the nasal-subcutaneous or nasal oral administration simultaneously with the basic therapy. Te studies were conducted first on limited groups of patients, then in the course of the State Trial with the use of placebo control. Excellent and good effect lasting for 1 year and over was registered in 36 patients (66.7%) out of 54 BA patients receiving the vaccine by the intranasal-subcutaneous method. Immunotherapy produced no positive effect in 13 patients (24.1%). Out of 35 examined patients receiving the vaccine by the intranasal oral method, excellent and good effect was registered in 26 patients (74.2%). No effect was registered in 4 patients (11.4%). In the group of 28 patients receiving placebo simultaneously with the basic therapy positive dynamics in the course of the disease was observed only in 3 patients. Treatment with polycomponent vaccine B[symbol: see text]-4 led to a prolonged (to a year and more) decrease in the frequency and severity of exacerbations, contributed to the prolongation of remissions and to a decrease in the amount of administered medicinal preparations, especially systemic corticosteroids. Immunotherapy ensured the correction of the content of lymphocyte subpopulations with markers CD3, CD4, CD72 and a rise in the titers of antibodies to antigens contained in the preparation. The use of therapeutic polycomponent vaccine B[symbol: see text]-4 for the treatment of patients with latex allergy ensured the state of prolonged remission in this group of patients. On the basis of our investigations we believe that the use of the therapeutic polycomponent vaccine B[symbol: see text]-4 may be included into the basis therapy of allergic diseases. PMID- 12630353 TI - [Vaccinal prophylaxis of hepatitis A in servicemen of the Far East military district]. AB - The importance of hepatitis A (HA) for epidemiological situations among the servicemen of the Armed Forces of the Russian Federation, including the Far Eastern Military District, and the necessity of vaccinal prophylaxis have been substantiated. Vaccination against HA, made in a single injection, was shown to be capable of decreasing morbidity, on the average, 1.98 times. The epidemiological effectiveness of vaccine "Avaxim" proved to be 89.6%. The results obtained in this investigation are indicative of the expediency of vaccination not only for prophylactic purposes, but also for localization and liquidation of epidemic foci of HA. PMID- 12630352 TI - [Immune response in adult immunized with adsorbed DT-M anatoxin with reduced antigen content and Imovax-DT-adulte vaccine]. AB - The comparative study of immune response after immunization of adults with adsorbed DT toxoid with reduced antigen content and Imovax-DT-adulte vaccine, as well as the safety of these preparations, was made. The study revealed that immunization of adults with adsorbed DT toxoid having reduced antigen content, made in two injections, and the injection of Imovax-DT-dulte vaccine, as well as the successive injection of these preparations, produced the same the levels of antitetanus immunity. Antidiphtheria immunity, evaluated by the number of seroconverted to diphtheria persons following two injections immunization was similar for the two preparations, while the level of antidiphtheria antibodies was higher in persons immunized with adsorbed DT toxoid. The immune stratum index was rather high among persons aged 16-29 years. This age group exhibited the highest number of persons, seropositive to both diphtheria and tetanus. Both vaccine preparations, adsorbed DT toxoid with reduced antigen content and Imovax DT-adulte vaccine, were found to be equally capable of inducing autoimmune reactions in the vaccinees, detected by laboratory methods. PMID- 12630354 TI - [Lectin of Yersinia pestis vaccine strain EV and its immunological properties]. AB - As the result of the chromatographic separation of Y. pestis EV membrane proteins, a protein fraction with hemagglutinating activity was obtained. The isolated preparation was glycoprotein with a molecular weight of 22 kD, contained 16% of carbohydrates and exhibited thermolabile properties. The determination of the carbohydrate specificity of this glycoprotein revealed that it belonged to the class of lectins. Changes in the content of 11 corticosteroids and the population composition of lymphocytes, as well as the detection of specific antibodies in the blood serum of guinea pigs immunized with lectin, were indicative of the fact that the preparation was sufficiently immunogenic and induced the activation of the processes of proliferation and activation of lymphocytes during immunogenesis. The lectin isolated from Y. pestis EV outer membrane may be regarded as an additional factor ensuring the contact of the pathogen with the cells of the body and as a promising component of combined plague vaccine. PMID- 12630355 TI - [Early diagnostics of hemorrhagic fever with renal syndrome on the basis of the use of Pumala virus recombinant nucleocapsid protein]. AB - Pumala virus recombinant nucleocapsid protein was used for the early diagnosis of haemorrhagic fever with renal syndrome. Specific IgM in the sera of patients could be determined by the IEA technique as early as on days 2-3 from the onset of the disease. The diagnostic effectiveness of the test-system was 95% and its specificity was 98%. PMID- 12630356 TI - [Parital microflora of human intestine]. AB - The modified method of the microbiological study of parietal mucin has been developed. The proposed method makes it possible to evaluate the parietal microflora of the intestine. The advantages of using physiological saline and Hanks' solution as medium ensuring the storage and self-thinning of mucin have been proved. The optimum time of making the microbiological study of intestinal mucin has also been determined. PMID- 12630358 TI - [Antibodies to meningococcal iron regulated protein FbpA and minor proteins in the sera of patients with meningococcal meningitis]. AB - Altogether 7 blood serum specimens taken from 3 patients with meningococcal meningitis were studied by the method of immunoblotting. The study revealed that on day 7 and especially on day 10 from the onset of the disease antibodies to periplasmatic iron-regulated protein FbpA with a molecular weight of 34 kD appeared in the serum of one patient. In the sera of two other patients the appearance of antibodies to minor iron-regulated proteins with molecular weights of 43 kD and 46 kD, absent at the acute stage of the disease, were detected on day 7. As the process of convalescence was progressing, in all patients under observation an increase in the specific immune response to proteins of class 5 with molecular weights of 20-14 kD was observed. PMID- 12630357 TI - [Neovir therapy in acute hepatitis C]. AB - Neovir, an inductor of interferon-alpha, was injected intramuscularly to 15 male patients aged 16-44 years with diagnosed acute viral hepatitis in a dose of 250 mg a day, 2-3 times a week, for 3 months. By the end of the therapy hepatitis C virus RNA could still be isolated in 84.6% of the patients; the values of AlaT, AsaT and alkaline phosphatase exceeded the normal levels twofold. The result of the therapy is regarded as ineffective. PMID- 12630359 TI - [Epidemiological and immunological monitoring of tetanus in Georgia (Republic)]. AB - During the period of 1961-1990 the regular immunological and epidemiological monitoring of tetanus was carried out. At the prevaccination period (1937-1960) 140 cases of tetanus were registered, 77 patients died (morbidity rate being 0.97 0.11 per 100,000 of the population with mortality rate equal to 55.6%). At the period of mass vaccinal prophylaxis (1960-1990) 56 cases were registered and 27 patients died (the corresponding values: 0.09-0.11 with 48.2% mortality). In 1981 1990 altogether 53,213 vaccinees were examined. 4,864 vaccinees (8.9%) proved to be seronegative and 43,615 vaccinees (81.9%) were found to have protective antibody titers. 4,220 pregnant women were examined; 476 women (11.2%) proved to be seronegative and 3,576 women (84.3%) were found to have protective antibody titers. Timely specific prophylaxis proved to be the only way for decreasing tetanus morbidity. PMID- 12630360 TI - [Disturbance of immune characteristics of the mucous membranes of the respiratory tract in bronchopulmonary diseases in children]. AB - A total of 100 children with bronchopulmonary diseases were examined. Of these, in 80 bronchial asthma and in 20 children acute pneumonia were diagnosed. The deficiency of local cell mediated and humoral factors on the mucous membranes of the upper respiratory tracts was established. In the squamous and columnar epithelium cells an increased destruction and cytolysis was noted leading to disturbances in the integrity of the protective barrier of the epithelial cover. In addition, this study revealed the deficiency of neutrophilic leukocytes, pronounced local eosinophilia and the low level of humoral protective factors. In acute pneumonia an increase in the amount of neutrophilic leukocytes in the secretion of the nasal cavity together with the appearance of destructive processes in epithelial cells took place. Pronounced deficiency in serum IgA and IgG was accompanied by compensatory increase in secretory IgA. PMID- 12630361 TI - [Effectiveness of the oral administration of tomicide in experimental infection]. AB - Experimental data on the oral administration of Tomicid using different schemes for the protection of white mice from staphylococcal infection are presented. The use of Tomicid, administered in the maximum dose admissible for mice, ensured the protection of up to 2/3 of the total number of mice. A single oral administration of the preparation immediately after infection protected 1/3 of the survived mice from local staphylococcal infection. Good prospects of using Tomicid for the prevention of catarrhal diseases in guinea pigs were established. PMID- 12630362 TI - [Mechanisms of the protective action of plant preparation erakond in influenza]. AB - The results of the study of some mechanisms of the protective action of Erakond, a phytopreparation intended for use in acute influenza infection are presented. In controlled animal observations Erakond was shown to induce the synthesis of antibodies in infected and immunized mice, the production of lymphokine, endogenic interferon; to increase the phagocytic and adhesive activity of mononuclears. The preparation facilitated the normalization of the process of the peroxide oxidation of lipids in mice and rendered a positive effect on the course of the inflammatory process in the lungs of mice at the final stage of influenza infection. PMID- 12630363 TI - [Use of the bioluminescent method for the determination of bacterial adenosinetriphosphate (ATP-metry) in microbiology]. AB - The attention of a wide circle of specialists has recently been attracted by different methods for rapid determination of pathogenic microorganisms in biological specimens, environmental objects and foodstuffs, as well as in cases of possible acts of bioterrorism. In this respect the bioluminescent method for determination of adenosine triphosphate (ATP) contained in microbial cells is of interest. The method is based on the interaction ATP, luciferase and luciferin, accompanied by giving off energy in the form of light emission. When compared with routine methods, the use of this method considerably reduces the duration of the analysis, and its high sensitivity is comparable with that of the polymerase chain reaction. In this review the data on the prospects of the practical use of the bioluminescent method of ATP-metry are presented. PMID- 12630364 TI - [Subtypes of human immunodeficiency virus 1:classification, the origin and circulation in Europe]. AB - The modern classification of human immunodeficiency viruses of type 1 (HIV-1) is presented. The hypotheses concerning the origin of this virus are discussed. The data on the spread of the subtypes of HIV-1 in different countries of Western and Eastern Europe and in different risk groups are presented. PMID- 12630365 TI - [Features of the epidemiology of modern cholera]. AB - In this review information on the spread of cholera in the world, embracing the period of the XIX and XX centuries, is presented. The detailed description of the pandemic character of cholera, including all seven pandemics, is given. The epidemiological situation in cholera is analyzed and the conclusion is made that the main regularity of the epidemic process in modern EI Tor cholera are the endemic character of the disease and its epidemic character due to the prolonged and continuous course of pandemic VII. The work points out that the epidemic process on the global level, as well as on the regional and local levels, is regulated mainly by growing activation of the international and internal movement of the population. PMID- 12630366 TI - [Tularemia in inactive natural foci]. AB - The data on the epidemiological and epizootological manifestations of tularemia on the territory of the Ulyanovsk region are presented. The characteristics describing the process of the of Francisella tularensis circulation as well as the environmental objects, most important for the manifestations of tularemia infection, are given. PMID- 12630367 TI - Get hip on bone health. PMID- 12630368 TI - Batting an eye at HIV. PMID- 12630369 TI - Tenofovir helps beat hepatitis B. PMID- 12630371 TI - Antibody therapy zaps cancer. PMID- 12630372 TI - Does HAART affect anal growths? PMID- 12630370 TI - Fighting heart disease with fish? PMID- 12630373 TI - Moving forward in HIV medicine. PMID- 12630374 TI - What's on the horizon? PMID- 12630375 TI - The seventh annual HIV drug guide. PMID- 12630376 TI - Drug prices. PMID- 12630377 TI - Behind the frontline. A doctor looks at combos & side effects. PMID- 12630378 TI - The ever-changing HIV treatment landscape. PMID- 12630379 TI - Combination dosing adjustments. PMID- 12630381 TI - Two cases of liver alveolar echinococcosis associated with simultaneous lung and brain metastases. PMID- 12630380 TI - Highly pyrimethamine-resistant alleles of dihydrofolate reductase in isolates of Plasmodium falciparum from Tanzania. AB - In 2000 we used a sensitive technique to examine 9 isolates from malaria patients in Muheza, Tanzania who had failed treatment with sulfadoxine-pyrimethamine (SP). Three isolates carried, at low levels, the leucine to isoleucine change at amino acid 164 that is associated with clinical failure of SP. Numerous other highly resistant alleles were also observed. PMID- 12630383 TI - Ph.D.'s as science teachers: notes from the field. PMID- 12630382 TI - Treating angina with EECP therapy. PMID- 12630384 TI - Motivational interviewing and dietary behavior change. PMID- 12630385 TI - Where can I find resources on food labeling in a restaurant setting? PMID- 12630386 TI - No right to assisted suicide. PMID- 12630387 TI - Recalcitrant cellulitis. PMID- 12630388 TI - [Effects of macrolide antibiotics on phagocytosis of apoptosis neutrophils by monocyte-derived macrophages]. PMID- 12630389 TI - Elimination of lymphatic filariasis: a public-health challenge. PMID- 12630390 TI - Observation unit protocol for fibrillation. PMID- 12630391 TI - Lymphatic filariasis elimination: progress in global programme development. PMID- 12630392 TI - Pediatric trauma is a surgical disease. PMID- 12630393 TI - Pediatric trauma is a surgical disease. PMID- 12630394 TI - Proper treatment of anaphylaxis. PMID- 12630395 TI - Routine screening for environmental radiation by first responders at explosions and fires. PMID- 12630396 TI - Resident work hours. PMID- 12630397 TI - Gun carrying among male adolescents as a function of gun ownership in the general population. PMID- 12630398 TI - Abstracts of the 1997 Annual Conference of the British Association of Sports and Exercise Sciences (BASES). PMID- 12630400 TI - Abstracts of the 2001 Meeting of the British Trauma Society. PMID- 12630399 TI - Abortion care for emergency physicians. PMID- 12630401 TI - Abstracts of the Society for Cardiovascular Magnetic Resonance 6th Annual Scientific Sessions. Lake Buena Vista, Florida, USA. February 7-9, 2003. PMID- 12630402 TI - Vector-control synergies, between 'roll back malaria' and the Global Programme to Eliminate Lymphatic Filariasis, in the African region. AB - The perspectives and opportunities for controlling the mosquito vectors of Wuchereria bancrofti in tropical Africa are summarized and discussed. The countries covered by the World Health Organization's African Region have about one third of the world's burden of lymphatic filariasis (LF) as well as large shares of the planet's malaria and of many other vector-borne diseases. African LF is entirely caused by nocturnally periodic W. bancrofti, filarial nematode that is transmitted in urban East Africa by Culex quinquefasciatus, and in rural areas across tropical Africa by the same anopheline species that transmit the Plasmodium spp. causing human malaria. The standard practices for controlling the vectors of malarial parasites - house-spraying with residual insecticides and the use of bednets (preferably pre-treated with insecticide) for personal and community protection - can be simultaneously effective against both LF and malaria. Although mass drug administrations remain the main stay of the current strategy for LF elimination, the vector-control activities of the 'Roll Back Malaria' campaign can be expected to reduce the transmission of W. bancrofti in co-endemic areas. The relevant issues of programme management and integrated vector control are briefly reviewed. PMID- 12630403 TI - Abstracts of the 8th Congress of ESACP, the 15th International Congress of ISDQP and the 6th Congress of the European Group of Telepathology. Heraclion, Crete, 14 21 September 2002. PMID- 12630404 TI - Abstracts of the 2nd International Society of Pediatric Oncology-- Aisa Conference and 6th National Conference of Pediatric Hematology-Oncology and Parents Meeting. November 22-24, 2002. New Delhi, India. PMID- 12630405 TI - [XVII Annual Meeting of Applied Research in Chagas Disease and V Annual Meeting of Applied Research in Leishmaniasis. Uberaba, Brazil, 25-28 October 2001]. PMID- 12630406 TI - Explosive events on the Sun. AB - I describe two of the most dynamic and highly energetic phenomena in the Solar System--the explosive flares that can occur when plasma is confined by magnetic fields and the large-scale ejections of material known as 'coronal mass ejections'. These explosive events are poorly understood and yet occur in a variety of contexts in the Universe, ranging from planetary magnetospheres to active galactic nuclei. Understanding why flares and coronal mass ejections occur is a major goal across a wide range of space physics and astrophysics. Although explosive events from the Sun have dramatic effects on Earth, flares in other stars, for example, can be vastly more energetic and have an even more profound effect on their environment. We are now in the unprecedented position of having access to a number of space observatories dedicated to the Sun: the Yohkoh spacecraft, the Solar and Heliospheric Observatory, the Transition Region and Coronal Explorer and the Ramaty High Energy Solar Spectroscopic Imager. These cover a wide wavelength range from white light to gamma rays with both spectroscopy and imaging, and allow huge progress to be made in understanding the processes involved in such large explosions. The high-resolution data show dramatic and complex explosions of material on all spatial scales on the Sun. They have revealed that the Sun is constantly changing everywhere on its surface- something that was never imagined before. One of the mechanisms that has been proposed to account for the large energy release is magnetic reconnection. Recent observations from space increasingly support this view. This article will discuss those observations that support this model and also those that suggest different processes. The current space missions have given us an excellent insight into the actual explosive processes in the Sun. However, they have provided us with only a tantalizing glimpse of what causes the elusive trigger. Future missions such as Solar-B (the follow-on to Yohkoh), the Solar Terrestrial Relations Observatory, the Solar Dynamics Observatory and the Solar Orbiter mission will allow us to probe the trigger in a way that was not dreamt of a decade ago, by providing stereo views, measurements from Sun-grazing orbit, and much higher spatial, temporal and spectral resolution. It is an exciting time for solar physics and everything that we learn about the Sun will improve our ability to understand other magnetic phenomena in the Universe. PMID- 12630407 TI - Are there gender differences in self-reported smoking practices? Correlation with thiocyanate and cotinine levels in smokers and nonsmokers from the Pawtucket Heart Health Program. AB - OBJECTIVES: This study compared serum cotinine and thiocyanate in assessment of self-reported smoking behavior among 1400 men and 1809 women from two New England communities. METHODS: Serum thiocyanate and serum cotinine levels were analyzed on 2411 and 798 survey respondents, respectively, in an attempt to provide an objective measurement for validation of self-reported smoking behaviors that were obtained through an in-home interviewer-administered questionnaire. Cross sectional household surveys were conducted with randomly selected men and women, aged 18-65, between 1981 and 1993 as part of the evaluation of the Pawtucket Heart Health Program. RESULTS: Among smokers, the thiocyanate test had similar rates of agreement for women(88.0%) and for men (89.3%). However, among nonsmokers, thiocyanate had higher rates of agreement for women (91.5%) than for men (85.2%). For cotinine, the rates of agreement among smokers were higher for women (91.6%) than for men (89.7%). Similarly, the rates of agreement among nonsmokers were also higher for women (93.9%) than for men (91.9%). Overall,serum cotinine had a higher concordance rate than serum thiocyanate for both men and women. CONCLUSIONS: Although our results suggested that there were some differences in self-reporting of smoking status by gender, results were quite similar between self-reports of smoking and both biochemical tests. The results obtained from this large population-based study from two New England communities lend credibility to the use of self-reports as a low-cost accurate approach to obtaining information on smoking behaviors among both men and women in large population-based health surveys. PMID- 12630408 TI - Genomes to Life "Center for Molecular and Cellular Systems": a research program for identification and characterization of protein complexes. AB - Goal 1 of Department of Energy's Genomes to Life (GTL) program seeks to identify and characterize the complete set of protein complexes within a cell. Goal 1 forms the foundation necessary to accomplish the other objectives of the GTL program, which focus on gene regulatory networks and molecular level characterization of interactions in microbial communities. Together this information would allow cells and their components to be understood in sufficient detail to predict, test and understand the responses of a biological system to its environment. The Center for Molecular and Cellular Systems has been established to identify and characterize protein complexes using high through-put analytical technologies.A dynamic research program is being developed that supports the goals of the Center by focusing on the development new capabilities for sample preparation and complex separations, molecular level identification of the protein complexes by mass spectrometry, characterization of the complexes in living cells by imaging techniques, and bioinformatics and computational tools for the collection and interpretation of data and formation of databases and tools to allow the data to be shared by the biological community. PMID- 12630409 TI - Vector-control synergies, between 'roll back malaria' and the Global Programme to Eliminate Lymphatic Filariasis, in South-east Asia. AB - The problems of, and opportunities for the control of filarial vectors in the World Health Organization's South-east Asian Region are summarized and discussed. Together, the countries in this region have more than half of the world's burden of lymphatic filariasis (LF), most of it caused by Wuchereria bancrofti transmitted by Culex quinquefasciatus. Although the vectors and the epidemiology and distribution of LF in this area largely differ from those of malaria, some vector-control methods (such as bednets and other personal-protection measures, breeding-site reduction and environmental management) can have an impact on LF, malaria and other mosquito-borne diseases. Some of the relevant issues of programme management and integrated vector control are reviewed. PMID- 12630410 TI - Pressure dependence of uveoscleral outflow. PMID- 12630411 TI - Closure of WISDOM and combined HRT arm of WHI. PMID- 12630413 TI - Nuclear cloning, stem cells, and genomic reprogramming. AB - The generation of adult animals by nuclear cloning from adult donor cells is extremely inefficient, with most clones dying soon after implantation. In contrast, cloning from embryonic stem cell donor nuclei is significanty more efficient than from adult donor cells. However, regardless of donor cell type, all clones that survive to birth and beyond suffer serious phenotypic and gene expression abnormalities. All available evidence is consistent with the notion that the anomalous phenotypes of cloned animals are caused by faulty epigenetic reprogramming of the donor nucleus. Faulty reprogramming appears to be caused by the cloning process itself as well as by the epigenetic state of the donor nucleus. In contrast to reproductive cloning, faulty reprogramming of the donor nucleus does not tend to interfere with the application of nuclear transfer technology for therapeutic purposes (therapeutic cloning). PMID- 12630412 TI - Multipotentiality of neuronal cells after spontaneous fusion with embryonic stem cells and nuclear reprogramming in vitro. AB - Primary mouse brain cells were cultured with HPRT (hypoxanthine phosphoribosyl transferase)-deficient ES (embryonic stem) cells to see if the ES cells could provide cues sufficient to reprogram a pluripotential state. After 5 days of coculture, HPRT-deficient ES cells were killed by selection in HAT (hypoxanthine, aminopterin, thymidine) medium. We observed islands of HAT-resistant ES-like cells surrounded by differentiated cells. Cell lines generated from three such "islands" proved to be spontaneous, pluripotential ES-neural hybrids, and gave rise to a chimera following blastocyst injection. Re-expression of the ES specific gene Foxd3 from somatic-derived chromosomes suggested that the somatic nucleus had been reprogrammed. Our results raise the intriguing possibility that ASCs shown to contribute to multiple tissues in blastocyst-injection studies may not contribute as a result of pluripotency. Instead contributions may arise from spontaneous fusion events in which phenotype is determined by either cytoplasmic dominance, nuclear reprogramming, or both. PMID- 12630415 TI - 3rd Yugoslav Coloproctology Symposium with International Participation. Belgrade, 11-12 October 2000. PMID- 12630414 TI - Position paper. Broadening the construct of independence. PMID- 12630416 TI - New toke on treating skin cancer. PMID- 12630418 TI - Viruses using their powers for good. PMID- 12630417 TI - Now hear this--scientists map the primate auditory system. PMID- 12630419 TI - Activities of daily living instruments:optimizing scales for neurologic assessments. AB - The ability to perform activities of daily living (ADL) is an important part of assessment in neurologic patients. A literature search was carried out to identify multi-item ADL scales developed for the assessment of neurologic patients, comparing item content, range, and detail of ADL scales. Of the 113 ADL scales identified, 27 (24%) were designed for use in neurology. In the basic ADL (BADL) domains (basic mobility and self-care), individual items were present in 44% to 81% of instruments. In the extended ADL (EADL) domains (e.g., outdoor mobility, housekeeping), items were present in up to 67% of the instruments identified. A typical trade-off was observed between the range, the detail (number of items), and hence the practicality of a scale. In general, scales focus on either BADL or EADL domains or, on occasion, some of both, rather than measuring the full range of functioning. There are many ADL scales in neurology, with much overlap in item content, leading to redundancy. New scales developed with the traditional methods will not solve the existing difficulties associated with range and detail, ordinal scale scores, and cross-instrument comparability. The possibilities of a modern psychometric method known as item response theory that was designed to solve these problems are discussed. PMID- 12630420 TI - Practice makes perfect. Interview by Alastair McLellan. PMID- 12630421 TI - e-novation. Pull the other one. PMID- 12630422 TI - e-novation. Under orders. PMID- 12630423 TI - e-novation. For your eyes only. PMID- 12630424 TI - e-novation. Band on the run. PMID- 12630425 TI - e-novation. A very secret service. PMID- 12630427 TI - SADA position paper on dental therapists--December 1999. PMID- 12630426 TI - [Immunoblastic lymphadenopathy-like T cell lymphoma with high level of serum interleukin-2 and various allergic reactions]. PMID- 12630428 TI - Dentists (small businesses) and the Basic Conditions of the Employment Act, 1977 (BCEA). PMID- 12630429 TI - Challenges in longitudinal designs in occupational health psychology. PMID- 12630430 TI - Factors influencing the impact of unemployment on mental health among young and older adults in a longitudinal, population-based survey. AB - OBJECTIVES: This study examined the relationship between unemployment and mental health. It particularly emphasized the potential differences in mental health status between younger workers entering the labor market and older workers with established laborforce involvement. METHODS: With the use of the National Population Health Survey in Canada, over 6000 respondents between 18 and 55 years of age in 1994 were followed up 2 years later. RESULTS: The results suggest that, among the 31- to 55-year-olds, becoming unemployed led to increases in distress and, to some extent, clinical depression at follow-up. This association between unemployment and mental health was not found among younger adults 18 to 30 years of age. Possible explanations for the null finding among young adults, such as decreased likelihood of low household income or increased likelihood of distressed young adults completely withdrawing from the workforce, were not supported. The notion that baseline mental health affects the chances of being unemployed at the time of a 24-month follow-up were partially supported. CONCLUSIONS: These findings from a representative sample suggest that both causation and selection processes lead to an association between unemployment and distress among older adults. PMID- 12630431 TI - Influence of lack of full-time employment on attempted suicide in Manitoba, Canada. AB - OBJECTIVES: Unemployment has been repeatedly associated with suicide; however, whether the association is causal remains unclear. Little is known about the relationship between part-time work and either attempted or completed suicide. The objective of this study was to compare the relationships of unemployment, part-time work, nonlaborforce participation, and full-time work with attempted suicide. METHODS: This study utilized a database consisting of 27446 potential laborforce participants that combines information on health care utilization in Manitoba, Canada, with detailed information from the 1986 census. Persons who attempted suicide after the census (N=144) were identified using established definitions based on hospital claims to identify serious attempts only. RESULTS: Step-wise multiple logistic regression, which controlled for multiple confounding variables, revealed that unemployment [odds ratio (OR) 3.68, 95% confidence interval (95% CI) 1.76-7.71, part-time work (OR 1.99, 95% CI 1.07-3.71) and being out of the labor force (OR 2.11, 95% CI 1.12-3.97)] were all associated with attempted suicide. A dose-response relationship was observed between weeks worked in 1985 and suicide attempts after the census. CONCLUSIONS: All three groups of those who were not working full-time had an elevated likelihood of attempted suicide after adjustment for potential confounding factors. This finding suggests that working full-time is protective against suicide attempts. Suicide attempts related to lack of full-time work may be more preventable than other causes of attempted suicide and may be decreased by social policies that limit "under" and unemployment. PMID- 12630432 TI - Timed bright-light exposure and complaints related to shift work among women. AB - OBJECTIVES: This field study measured whether repeated, brief exposures to bright light during night shifts improved subjective well-being during and after night work. A secondary objective was to investigate whether this response differed by season (summer or winter), seasonality, or age. METHODS: Eighty-seven healthy female nurses were voluntarily exposed to brief periods (4 x 20 minutes) of bright (5000 lux) light at scheduled times during every night shift over a 2-week period. Each morning following a night shift the subjects filled out self assessment questionnaires measuring subjective symptoms and distress caused by work at night. The questionnaires were also completed 2 weeks before and after the light intervention. The study had two phases, summer (May-June) and winter (November-December). Thirty-seven of the subjects participated during both periods. RESULTS: Light significantly alleviated the subjective distress associated with nightshift work, both in summer and in winter, independent of the subject's age. The effect was stronger for those who reported routine seasonal changes in mood. CONCLUSIONS: Short pulses of timed bright-light exposure may enhance subjective adaptation to night work. PMID- 12630433 TI - A shorter workday as a means of reducing the occurrence of musculoskeletal disorders. AB - OBJECTIVES: The study examined the relation between daily workhours and the occurrence of neck-shoulder or back pain in physically demanding care work. METHODS: Unpublished data were obtained from three intervention projects in care institutions. The projects had been conducted independently in Oslo (46 participants, 175 referents before and 158 referents after the intervention), Helsingborg (60 participants, 89 referents) and Stockholm (41 participants, 22 referents) between 1995 and 1998. The intervention was a reduction of daily workhours from > or = 7 to 6 hours (or 30 hours weekly). Full-time salary was retained, and extra personnel were employed to compensate for the reduction in workhours. Data were collected by self-administered questionnaires before and during the intervention periods, lasting from 12 to 22 months. RESULTS: The prevalence of neck-shoulder pain decreased from 40.9% to 25.6% in Oslo and from 57.1% to 39.1% in Helsingborg after 1.5 years with a 6-hour workday; for Stockholm the decrease was from 81.6% to 68.3% after 1 year. No decrease was observed in the reference groups. The prevalence of back pain did not show the same consistent pattern. CONCLUSIONS: The shortening of regular workdays from > or = 7 hours to 6 hours may considerably reduce the prevalence of neck-shoulder pain among persons with physically demanding care work. The potential health benefits should encourage intervention studies also in other occupations with increased risk of work-related musculoskeletal disorders. PMID- 12630434 TI - Risk factors predicting hip pain in a 5-year prospective cohort study. AB - OBJECTIVES: The aim of the study was to identify and quantify risk factors for hip pain. METHODS: A representative sample of 5001 Danish men and women aged 18 65 years in 1990 were interviewed about occupational exposures (response rate 90%); 5 years later they were reinterviewed about hip pain (response rate 86%). Logistic regression with forced entry of all the independent variables was used to estimate the odds ratios for the possible risk factors. The impact of the various predictors was assessed through the calculation of population etiologic fractions. RESULTS: A double risk of hip pain was found for the women as compared with the men [odds ratio (OR) 2.28, 95% confidence interval (95% CI) 1.68-3.09]. The risk increased with body mass index. Whole-body vibration (OR 1.86, 95% CI 1.09-2.71) and physically demanding work (OR 1.83, 95% CI 1.23-2.71) were strong predictors of hip pain, while a squatting work posture was protective (OR 0.64, 95% CI 0.42-0.98). The impact of the statistically significant predictors (the etiologic fractions) was as follows: 0.49 for body mass index, 0.05 for whole body vibration, 0.10 for physically demanding work, and 0.32 for squatting (preventive). CONCLUSIONS: Female gender, age, high body mass index, whole-body vibration, and physically demanding work are significant risk factors for hip pain. PMID- 12630435 TI - Annoyance and performance during the experimental chemical challenge of subjects with multiple chemical sensitivity. AB - OBJECTIVES: This study explored the subjective reactions and psychological test performance of smell-intolerant subjects during consecutive challenges to chemicals with contrasting neurotoxic properties. METHODS: Women with symptoms compatible with multiple chemical sensitivity (N=10) and healthy referents (N=20) were individually challenged in an exposure chamber. All the subjects attended two separate 2-hour sessions of exposure to n-butyl acetate and toluene, in counterbalanced sequence. After an initial phase without exposure, air concentrations were increased in steps ranging from 3.6 to 57 mg/m3 for n-butyl acetate and from 11 to 180 mg/m3 for toluene. The response measures comprised ratings of annoyance and smell intensity and also neurobehavioral test performance. RESULTS: Both groups showed an increase in annoyance ratings and a decrease in test performance in the initial unexposed chamber phase and also in the first phase of the chemical exposure, these results indicating slight immediate expectancy or "suggestion" effects. During the six chamber phases, the ratings of mucous membrane irritation and fatigue showed a steeper increase in the group with multiple chemical sensitivity than among the referents, while the ratings of smell intensity and smell annoyance were similar in the two groups. A reduction in test performance was observed during the chamber phases, particularly in the group with multiple chemical sensitivity. No relation was found between the ratings or performance and chemical substance. CONCLUSIONS: Stronger immediate expectancy or "suggestion" reactions than normal did not characterize the group with multiple chemical sensitivity. This group showed a stronger than normal gradual build-up of fatigue, mucous membrane irritation, and reduced performance during chemical exposure. The results offer the most support to an irritative basis for multiple chemical sensitivity. PMID- 12630436 TI - Exposure to benzene and risk of leukemia among shoe factory workers. AB - OBJECTIVES: The study attempted to add years of follow-up to an earlier study describing excess leukemia among workers exposed to benzene-based glues in a shoe factory and to conduct a quantitative exposure assessment of the exposure to benzene and the risk of leukemia. METHODS: The cohort comprised 1687 persons with complete work histories, at work on 1 January 1950, and followed through 31 December 1999. For each subject, time-specific cumulative exposure (ppm-years) was calculated as the sum of the products of job-specific concentrations of benzene (ppm) and the duration (years) for each job. Standardized mortality ratios (SMR) were estimated using national and regional rates specific for gender, age, and period. RESULTS: The cumulative exposure ranged from 0 to >500 ppm-years. The SMR values for all hematolymphopoietic malignancies and leukemia for the men and women combined were elevated in all but the lowest exposure category. Leukemia risk was significantly elevated in the highest exposure category and the most evident among the men. The SMR values for the men were 1.4 [95% confidence interval (95% CI) 0.2-5.0], 3.7 (95% CI 0.1-20.6), 3.0 (95% CI 0.4-10.9), and 7.0 (95% CI 1.9-18.0) for benzene, the cumulative exposure equaling <40, 40-99, 100-199, and >200 ppm-years, respectively. CONCLUSIONS: The results indicate that leukemia mortality is associated with exposure to benzene in this factory and that the risk increases with increasing cumulative exposure. The relevance of these findings for estimating risk at much lower levels is limited because of the small study size. PMID- 12630437 TI - Diffuse pleural fibrosis--an unreliable indicator of heavy asbestos exposure? AB - OBJECTIVES: This study aimed at testing the hypothesis that diffuse pleural fibrosis is associated with a heavier exposure to asbestos than that of benign pleural plaques, and hence diffuse pleural fibrosis can be used as a marker of heavy exposure. METHODS: Asbestos burden was estimated for 192 naval dockyard workers (96 with diffuse pleural fibrosis, 96 with plaques) by calculating the exposure ratings from the trade and the years spent in that trade. In 53 cases the diffuse pleural fibrosis was bilateral. RESULTS: No difference was found in the mean estimated asbestos burden between all diffuse pleural fibrosis and plaques, nor between plaques and unilateral and bilateral diffuse pleural fibrosis, analyzed separately by a one-factor analysis of variance. The mean asbestos burden was significantly greater for bilateral than for unilateral diffuse pleural fibrosis. A wide spread of asbestos exposure was noted among the men with diffuse pleural fibrosis. CONCLUSIONS: Diffuse pleural fibrosis may arise after widely varying exposure to asbestos. The mean exposure ratings for diffuse pleural fibrosis do not differ from those for pleural plaques, although the ratings are significantly higher for men with bilateral diffuse pleural fibrosis than for those with unilateral diffuse pleural fibrosis. Diffuse pleural fibrosis cannot be used as a reliable marker of heavy asbestos exposure. PMID- 12630438 TI - Incidence of cancer among the participants of the Finnish Asbestos Screening Campaign. AB - OBJECTIVES: Cancer risk has been estimated for asbestos production workers or other heavily exposed asbestos workers in numerous studies. The bulk of the asbestos epidemic results come, however, from past intermittent exposures during asbestos product use. This study concentrated on estimating the risk of cancer in such a population. METHODS: Altogether 23285 men and 930 women invited to a nationwide screening campaign for benign asbestos-related diseases in 1990-1992 were followed for cancer through the Finnish Cancer Register up to 1998. Standardized incidence ratios (SIR) were calculated in comparison with the total Finnish population. RESULTS: Altogether 1392 cases of cancer were found among the men. The risk was slightly, but significantly elevated for lung cancer [SIR 1.14, 95% confidence interval (95% CI) 1.01-1.26), mesothelioma (SIR 2.77, 95% CI 1.66 4.31), and prostate cancer (SIR 1.21, 95% CI 1.09-1.34). The risk of lung cancer was slightly higher among the invited nonparticipants (SIR 1.48, 95% CI 1.20 1.79) than among the participants (SIR 1.02, 95% CI 0.88-1.17). About 98% of the lung cancers occurred in current or ex-smokers. CONCLUSIONS: In a population of long-term construction workers, the risk of lung cancer and mesothelioma was increased, but considerably lower than among insulators, asbestos sprayers, or patients with asbestosis. As it was not possible to follow most of the invited nonparticipants in the original screening study, selection bias by smoking or other life-style factors possibly correlated to the individual's decision to participate in the health screening cannot be excluded. PMID- 12630439 TI - Prolonged respiratory symptoms caused by thermal degradation products of freons. AB - OBJECTIVES: The chlorofluorocarbons (CFC) used in refrigeration systems decompose on heating and produce substances that are highly irritating to the airways (eg, chlorine, carbonyl fluoride, and hydrogen fluoride). This study examined persistent respiratory symptoms among several workers exposed to thermal decomposition products of CFC. METHODS: Seven patients with respiratory symptoms caused by inadvertent exposure to thermal decomposition products of CFC in a restaurant kitchen or during refrigerator repair were studied with the use of spirometry, peak flow follow-up, and histamine challenge tests. Three patients also underwent bronchoscopy and bronchoalveolar lavage. RESULTS: In five of the cases, cough or dyspnea lasted longer than 1 month; for three of the five, the symptoms lasted more than 4 years. Three cases showed increased bronchial hyperreactivity, and two of the three had increased diurnal peak flow variation. Three patients fulfilled the criteria for acute irritant-induced asthma or reactive airway dysfunction syndrome. One case exhibited bronchiolitis while, for the other six, the clinical picture was consistent with bronchitis. CONCLUSIONS: The studied cases indicate that the thermal decomposition products of CFC used in refrigerators may cause irritant-induced airway diseases of long duration. PMID- 12630440 TI - Evidence-based medicine and evidence-based occupational health. PMID- 12630441 TI - Quality of evidence. PMID- 12630442 TI - Where to go from here? PMID- 12630443 TI - More evidence that herbicides feminize amphibians. PMID- 12630444 TI - NRC calls for new approach to livestock emissions. PMID- 12630445 TI - NSF budget could double over five years. PMID- 12630446 TI - High PAH levels in dust from 9-11 disaster. PMID- 12630447 TI - Real-time PCR monitors wastewater treatment. PMID- 12630448 TI - Pesticide mixture enhances frog abnormalities. PMID- 12630449 TI - Iodine counteracts perchlorate effects in frogs. PMID- 12630450 TI - Barring aquatic invaders. PMID- 12630451 TI - EC proposes sulfur limits on marine fuels. PMID- 12630452 TI - Southeast Asia development raises concerns. PMID- 12630453 TI - Small fiber estimates body residues. PMID- 12630454 TI - Taking credit for forest carbon sinks. PMID- 12630455 TI - Theoretical ecology for engineering biology. PMID- 12630456 TI - Origin of lead in eight Central European peat bogs determined from isotope ratios, strengths, and operation times of regional pollution sources. AB - Lead originating from coal burning, gasoline burning, and ore smelting was identified in 210Pb-dated profiles through eight peat bogs distributed over an area of 60,000 km2. The Sphagnum-dominated bogs were located mainly in mountainous regions of the Czech Republic bordering with Germany, Austria, and Poland. Basal peat 14C-dated at 11,000 years BP had a relatively high 206Pb/207Pb ratio (1.193). Peat deposited around 1800 AD had a lower 206Pb/207Pb ratio of 1.168-1.178, indicating that environmental lead in Central Europe had been largely affected by human activity (smelting) even before the beginning of the Industrial Revolution. Five of the sites exhibited a nearly constant 206Pb/207Pb ratio (1.175) throughout the 19th century, resembling the "anthropogenic baseline" described in Northern Europe (1.17). At all sites, the 206Pb/207Pb ratio of peat decreased at least until 1980; at four sites, a reversal to more radiogenic values (higher 206Pb/207Pb), typical of easing pollution, was observed in the following decade (1980-1990). A time series of annual outputs for 14 different mining districts dispersing lead into the environment has been constructed for the past 200 years. The production of Ag-Pb, coal, and leaded gasoline peaked in 1900, 1980, and 1980, respectively. In contrast to other European countries, no peak in annual Pb accumulation rates was found in 1900, the year of maximum ore smelting. The highest annual Pb accumulation rates in peat were consistent with the highest Pb emission rates from coal-fired power plants and traffic (1980). Although maximum coal and gasoline production coincided in time, their isotope ratios were unique. The mean measured 206Pb/207Pb ratios of local coal, ores, and gasoline were 1.19, 1.16, and 1.11, respectively. A considerable proportion of coal emissions, relative to gasoline emisions, was responsible for the higher 206Pb/207Pb ratios in the recent atmosphere (1.15) compared to Western Europe (1.10). As in West European countries, the gasoline sold in the Czech Republic during the Communist era (1948 1989) contained an admixture of low-radiogenic Precambrian lead from Australia. PMID- 12630457 TI - Air quality model evaluation data for organics. 6. C3-C24 organic acids. AB - The atmospheric concentrations of 47 carboxylic acids in the semivolatile and particle phases are quantified in the Los Angeles area, as part of a larger study of the vapor-phase, semivolatile, and particle-phase organic compounds. Variations in the spatial and temporal distributions of acid concentrations are analyzed to determine whether atmospheric formation or primary emissions are responsible for the observed levels. Relatively low molecular weight aliphatic dicarboxylic acids (e.g., butanedioic acid, hexanedioic acid, and propanedioic acid) and some n-alkanoic acids (e.g., n-octanoic acid and n-nonanoic acid) are found at an offshore sampling location at levels comparable to urban area concentrations indicating that these compounds or their atmospheric precursors may be derived from long-range transport or natural background sources. Some aromatic carboxylic acids (e.g., benzoic acid and 1,2-benzenedicarboxylic acid) have spatial and temporal distributions suggesting that formation from anthropogenic emissions of gaseous precursors dominates their atmospheric concentrations. Additionally, the distributions of aliphatic carboxylic acid concentrations known to be emitted from primary sources (e.g., hexadecanoic acid and octadecanoic acid) are consistent with direct emissions as the dominant source of these compounds. PMID- 12630458 TI - Spatial and temporal trends of POPs in Norwegian and UK background air: implications for global cycling. AB - Data are presented for PCBs and HCB measured by passive air samplers (SPMDs) along a latitudinal transect from the south of the UK to the north of Norway during 1998-2000. This work is part of an ongoing air sampling campaign in which data were previously gathered for 1994-1996. Comparisons of the masses of chemicals sequestered by the SPMDs during these different time intervals are used to investigate spatial and temporal trends. Results are discussed in the context of sources, long-range atmospheric transport, fractionation/cold condensation, and global clearance processes controlling ambient levels of POPs. Spatial trends show a decrease in absolute sequestered amounts of PCBs with increasing latitude i.e., with increasing distance from the source area. However, relative sequestered amounts of the homologue groups (expressed as a ratio to penta-PCB) show a clear latitudinal trend, with the relative contribution of the lighter congeners increasing with increasing latitude, providing evidence of latitudinal fractionation. Absolute amounts of HCB increase with latitude, suggesting this compound is undergoing cold condensation. Sequestered amounts of PCBs generally decreased between the two sampling periods by a factor 2-5 over 4 years, suggesting half-lives on the order of 1.7-4 years. The relative rates of decline (1998-2000 data as a percentage of the 1994-1996 data) were compared for different congeners and latitudes. No clear latitudinal trends were found, with all sites/congeners showing a similar marked decline over time to ca. 30% of the former value. We discuss the interpretation of these observations and conclude they imply that the underlying trends of current ambient levels of PCBs in European background air are still largely controlled by primary emissions, rather than recycling/secondary emissions from the major environmental repositories such as soils or water bodies. PMID- 12630459 TI - PAHs, PCBs, PCNs, organochlorine pesticides, synthetic musks, and polychlorinated n-alkanes in U.K. sewage sludge: survey results and implications. AB - A survey of the digested sludge from 14 U.K. wastewater treatment plants was carried out to obtain contemporary U.K. data on the concentrations of certain classes of persistent organic compounds for which data are scarce and to assess whether U.K. sludge was likely to comply with the sludge limits for PCBs and PAHs proposed by the European Union. Total PAH (24 compounds) concentrations ranged from 67 to 370 mg/kg dw, in line with data from other countries; all the samples would exceed the proposed EU limit. Total PCB concentrations were 110-440 microg/kg dw, well below the proposed EU limit. Total PCN concentrations ranged from 50 to 190 microg/kg. Total synthetic musk concentrations ranged from 2.1 to 86 mg/kg dw; there were a few very high concentrations of HHCB and AHTN in the samples. Total concentrations of the short- and medium-chained polychlorinated alkanes ranged between 7 and 200 mg/kg and between 30 and 9700 mg/kg, respectively. These very high concentrations are indicative of chemicals with numerous and ongoing diffuse sources. PMID- 12630460 TI - Polychlorinated dibenzo-p-dioxin and dibenzofuran concentration profiles in sediments and flood-plain soils of the Tittabawassee River, Michigan. AB - Concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in sediments and flood-plain soils collected along the Tittabawassee River in Michigan ranged from 102 to 53,600 pg/g, dry wt. Mean PCDD/PCDF concentrations in downstream sediment and soil were from 10- to 20-fold greater than those found at locations upstream of Midland, Michigan. Concentrations of PCDD/PCDF in sediments and flood-plain soils from the Tittabawassee watershed were comparable to those found in industrialized areas such as the Housatonic and lower Passaic Rivers in the U.S. Concentrations of PCDDs/PCDFs in soil and sediment were not correlated with total organic carbon (TOC) in sediments or soils. OCDD and 2,3,7,8-TeCDF were the predominant congeners in sediment/soil collected from locations downstream of Midland, Michigan. Principal component analysis of the PCDD/PCDF congener profile suggested the presence of sources originating from a mixture of chlorophenol and other chlorinated compound production. Mass balance analysis of TCDD equivalents (TCDD-EQs) derived from H4IIE-luc bioassay of sediment extracts and 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQs) estimated from instrumental analysis suggested that PCDDs/PCDFs were the major dioxin-like compounds present in sediments. A significant correlation existed between bioassay-derived TCDD-EQs and instrumentally measured TEQs (r2 = 0.94). PMID- 12630461 TI - Radial dilution model for the distribution of toxaphene in the United States and Canada on the basis of measured concentrations in tree bark. AB - Toxaphene was a highly chlorinated pesticide that was used in the United States from the 1950s until its restriction and ban in the 1980s. It was primarily used on cotton in the southern United States, and perhaps as a result, toxaphene is found in high concentrations in the southern United States. Toxaphene has also been detected at remote locations, such as in the high Canadian Arctic. However, these and other studies focused on selected regions of the United States and Canada, which only allowed for a limited interpretation of these data. We report here the concentrations of toxaphene measured in 46 tree bark samples collected in the United States and Canada. Concentrations were found to be greatest in the Mississippi River Valley along the borders of southern Missouri and Arkansas. We created a simple model based on radial dilution from a central location in the south central United States to describe toxaphene concentrations in tree bark and in air throughout the United States and Canada. The toxaphene concentrations were successfully described by this inverse square distance model. A bark-atmosphere partition coefficient for toxaphene was also calculated that was similar to literature-derived octanol-air partition coefficients. High concentrations of toxaphene still exist in areas where it was heavily used but decline rapidly with distance. PMID- 12630462 TI - Diffusion coefficients of humic substances in agarose gel and in water. AB - Measurements of the diffusion coefficients of five different humic substances (HS) have been performed in water and in agarose hydrogels at several pH values (in the range of 3-10) and gel concentrations (in the range of 0.7-3% w/w). Fluorescence correlation spectroscopy (FCS) and classical diffusion cells were used in parallel to probe diffusion over both microscopic and mesoscopic distance scales. In general, agreement between the techniques was reasonable, which indicated that local nonhomogenities in the gel did not play an important role. Diffusion coefficients (D) in the gel were generally in the range of 0.9-2.5 x 10(-10) m2 s(-1) but were generally only 10-20% lower than in solution. At low pH values, one of the studied humic substances (a peat humic acid, PPHA) formed large aggregates that could not penetrate into the gel and therefore could not be defined by a single D value. The observed decreases of D in the gel for other HS were too large to be explained by the tortuousity and obstructive effects of the gel alone. D decreased slightly with increasing gel concentration and increased slightly with pH. Because modifications of D due to pH were similar in both the gel and the free solution, it is unlikely that complexation with the gel was greatly influenced by the pH. Rather, the main effect that appeared to decrease the diffusive flux in gels was likely small increases in the hydrodynamic radii of the humic macromolecules. An anomalous diffusion model was used to describe the FCS data in the gel. The characteristic exponent determined by fitting the autocorrelation functions with this model decreased only slightly (from 0.96 to 0.90) with increasing gel concentration providing support that HS complexation with the gel fibers was not very important. The results have important implications for our understanding of the fate and behavior of the HS and their associated pollutants and for interpreting metal speciation data obtained using gel-covered analytical sensors. PMID- 12630463 TI - DDT-related compounds bound to the nonextractable particulate matter in sediments of the Teltow Canal, Germany. AB - Sediment samples of the Teltow Canal (Berlin, Germany) were analyzed with respect to extractable and nonextractable organic compounds. The study focused on the identification and quantitation of bound 2,2-bis(chlorophenyl)-1,1,1 trichlorethane (DDT) residues in order to obtain further information about the fate of DDT-derived compounds within the particulate matter of the aquatic environment. Various chemical degradation techniques and a complementary online pyrolysis-GC/MS method were applied to the pre-extracted sediment residues. Generally, the distribution of the bound DDT-related compounds was found to differ distinctly from the substances distribution within the extractable fraction. The main metabolite of the anaerobic degradation pathway (2,2 bis(chlorophenyl)-1,1-dichlorethane, DDD) is most abundant in the sediment extracts but occurred only in insignificant concentrations in the degradation products of all procedures applied. The most abundant DDT-metabolites released after the degradation procedures were 4,4'-DBP, 4,4'-DDA, and 4,4'-DDM. In addition, 4,4'-DDM was detected at rather high concentrations by pyrolytic analysis. The results imply a weak association to the nonextractable particulate matter based on noncovalent interactions for the observed DDT-related contaminants. The release of these compounds was initiated by the modification and degradation of the organic macromolecular matrix as well as of the inorganic material. Furthermore, numerous methoxychlor-related compounds were detected not only in the extracts but also in parts of the hydrolysis products. PMID- 12630464 TI - Characterizing geochemical reactions in unsaturated mine waste-rock piles using gaseous O2, CO2, 12CO2, and 13CO2. AB - In situ determinations of geochemical reaction rates in mine waste-rock piles remain a challenge. Depth-profiles of field O2 and CO2 pore-gas concentrations, delta13C(CO2) values, and moisture contents were used to characterize and quantify geochemical reaction rates in two waste-rock piles at the Key Lake Uranium Mine in northern Saskatchewan, Canada. Traditionally, the presence of O2 concentrations less than atmospheric in waste-rock piles has been attributed to mineral oxidation. This study showed that the interpretation of O2 and CO2 concentration profiles alone could not be used to identify the depths of dominant geochemical reactions in the piles and could lead to erroneous estimates of reaction rates. Modeling of the delta13C(CO2) depth profiles clearly showed that the gas concentration profiles present in the piles were the result of the oxidation of organic matter present below the piles, a mechanism not previously reported in the literature. Based on these findings, the rates of reactions in the organic zone were determined. The oxidation of organic matter at the base of waste-rock piles should be considered in future mine-waste pore-gas studies, in addition to sulfide oxidation and carbonate buffering. PMID- 12630466 TI - Kinetically inert Cu in coastal waters. AB - Many studies have shown that Cu and other metals in natural waters are mostly bound by unidentified compounds interpreted to be strong ligands reversibly complexing a given metal. However, commonly applied analytical techniques are not capable of distinguishing strongly but reversibly complexed metal from metal bound in kinetically inert compounds. In this work, we use a modified competitive ligand exchange adsorptive cathodic stripping voltammetry method combined with size fractionation to show that most if not all of the apparently very strongly (log K > or = 13) bound Cu in samples from five New England coastal waters (1-18 nM, 10-60% of total Cu) is actually present as kinetically inert compounds. In three of the five samples examined by ultrafiltration, a significant portion of the 0.2-microm-filtrable inert Cu was retained by a 0.02-microm-pore size filter, suggesting that at least some of the Cu was kinetically inert because it was physically sequestered in colloidal material. The rest of the ambient Cu, and Cu added in titrations, were reversibly bound in complexes that could be modeled as having conditional stability constants of 10(10)-10(13). The Cu-binding ability of these complexes was equivalent to that of seawater containing reasonable concentrations of humic substances from terrestrial sources, approximately 0.15 0.45 mg of C/L. Both the inert compounds and the reversible ligands were important for determining [Cu2+] at ambient Cu levels in our samples. PMID- 12630465 TI - Persistent organic pollutants in the dusts that settled across lower Manhattan after September 11, 2001. AB - The explosion and collapse of the World Trade Center (WTC) was a catastrophic event that produced an aerosol impacting many workers, residents, and commuters during the first few days after September 11, 2001. During the initial days that followed, 14 bulk samples of the settled dust were collected at locations surrounding the epicenter of the disaster, including one indoor location. Some samples were analyzed for many potential hazards, including inorganic and organic constituents as well as morphology. The results of the analyses for persistent organic pollutants are described herein, including polycyclic aromatic hydrocarbons, polychlorinated biphenyls, and select organochlorine pesticides on settled dust samples. The sigma86-PCBs comprising less than 0.001% by mass of the bulk in the three bulk samples analyzed indicated that PCBs were of limited significance in the total settled dust across lower Manhattan. Likewise, organochlorine pesticides, including chlordanes, hexachlorobenzene, heptachlor, 4,4'-DDE, 2,4'-DDT, 4,4'-DDT, and Mirex, were found at low concentrations in the bulk samples. Conversely, the sigma37-PAHs comprised up to nearly 0.04% (<0.005 0.039%) by mass of the bulk settled dust in the six bulk samples. Further size segregation of these three initial bulk samples and seven additional samples indicates that sigma37-PAHs were found in higher concentrations on relatively large particles (10-53 microm), representing up to 0.04% of the total dust mass. Significant concentrations were also found on fine particles (<2.5 microm), often accounting for approximately 0.005% by mass. We estimate that approximately 100 1000 tons of sigma37-PAHs were spread over a localized area immediately after the WTC disaster on September 11. PMID- 12630468 TI - Nickel sequestration in a kaolinite-humic acid complex. AB - Incorporation of first row transition metals into stable surface precipitates can play an important role in reducing the bioavailability of these metals in neutral and alkaline soils. Organic coatings may interfere with this sorption mechanism by changing the surface characteristics and by masking the mineral surface from metal sorptives. In this study, kinetic sorption and desorption experiments were combined with extended X-ray absorption fine structure (EXAFS) spectroscopy to elucidate the effect of humic acid (HA) coatings on the formation and stabilization of nickel precipitates at the kaolinite-water interface. Initial Ni uptake (pH 7.5, [Ni]i = 3 mM, and I = 0.02 M NaNO3) increased with greater amounts of HA coated onto the kaolinite surface. Ni uptake continued over an extended period of time without reaching an apparent equilibrium. EXAFS analysis of the Ni sorption complex structures formed over time (up to 7 months) revealed the formation of a Ni-Al layered double hydroxide (LDH) precipitate at the kaolinite surface in the absence of HA. HA alone formed an inner-sphere complex with Ni (with 2 carbon atoms at an average radial distance of 2.85 A). A Ni-Al LDH precipitate phase was formed at the kaolinite surface in the presence of a 1 wt % HA coating. However, with 5 wt % HA coated at the kaolinite surface, the formation of a surface precipitate was slowed significantly, and the precipitate formed was similar in structure to Ni(OH)2(s). The Ni(OH)2 precipitate was not resistant to proton dissolution, while the Ni-Al LDH precipitate was. These results augment earlier findings that the incorporation of Ni and other first row transition metals into stable surface precipitates is an important sequestration pathway for toxic metals in the environment, despite the presence of ubiquitous coating materials such as humic acids. PMID- 12630467 TI - Reactivity of Fe(II) species associated with clay minerals. AB - Mineral-bound Fe(II) species represent important natural reductants of pollutants in the anaerobic subsurface. At clay minerals, three types of Fe(II) species in fundamentally different chemical environments may be present simultaneously, i.e., structural Fe(II), Fe(II) complexed by surface hydroxyl groups, and Fe(II) bound by ion exchange. We investigated the accessibility and reactivity of these three types of Fe(II) species in suspensions of two different clay minerals containing either ferrous iron-bearing nontronite or iron-free hectorite. Nitroaromatic compounds (NACs) exhibiting different sorption behavior on clays were used to probe the reactivity of the various types of reduced iron species. The clay treatment allowed for a preparation of nontronite and hectorite surfaces with Fe(II) adsorbed by surface hydroxyl groups at the edge surfaces. Furthermore, hectorite suspensions with additional Fe(II) bound to the ion exchange sites at the basal siloxane surfaces were set up. We found that both structural Fe(II) and Fe(II) complexed by surface hydroxyl groups of nontronite reduced the NACs to anilines. An electron balance revealed that more than 10% of the total iron in nontronite was reactive Fe(II). Fe(II) bound by ion exchange did not contribute to the observed reduction of NACs. Reversible adsorption of the NACs at the basal siloxane surface of the clays strongly retarded NAC reduction, even in the presence of high concentrations of Fe(II) bound by ion exchange to the basal siloxane surfaces. Our work shows that in natural systems a fraction of the total Fe(II) present on clays may contribute to the pool of highly reactive Fe(II) species in the subsurface. Furthermore, this work may help to distinguish between Fe(II) species of different reactivity regarding pollutant reduction. Although structural iron in clays represents only a small fraction of the total iron pool in soils and aquifers, reactive Fe(II) species originating from the reduction of structural Fe(III) in clays may contribute significantly to the biogeochemical cycling of electrons in the subsurface since it is not subject to depletion by reductive dissolution. PMID- 12630469 TI - Reductive capacity of natural reductants. AB - Reductive capacities of soil minerals and soil for Cr(VI) and chlorinated ethylenes were measured and characterized to provide basic knowledge for in-situ and ex-situ treatment using these natural reductants. The reductive capacities of iron-bearing sulfide (pyrite), hydroxide (green rust; GR(SO4)), and oxide (magnetite) minerals for Cr(VI) and tetrachloroethylene (PCE) were 1-3 orders of magnitude greater than those of iron-bearing phyllosilicates (biotite, vermiculite, and montmorillonite). The reductive capacities of surface soil collected from the plains of central Texas were similar and slightly greater than those of iron-bearing phyllosilicates. The reductive capacity of iron-bearing soil minerals for Cr(VI) was roughly 3-16 times greater than that for PCE, implying that Cr(VI) is more susceptible to being reduced by soil minerals than is PCE. GR(SO4) has the greatest reductive capacity for both Cr(VI) and PCE followed by magnetite, pyrite, biotite, montmorillonite, and vermiculite. This order was the same for both target compounds, which indicates that the relative reductive capacities of soil minerals are consistent. The reductive capacities of pyrite and GR(SO4) for chlorinated ethylenes decreased in the order: trichloroethylene (TCE) > PCE > cis-dichloroethylene (c-DCE) > vinyl chloride (VC). Fe(II) content in soil minerals was directly proportional to the reductive capacity of soil minerals for Cr(VI) and PCE, suggesting that Fe(II) content is an important factor that significantly affects reductive transformations of target contaminants in natural systems. PMID- 12630470 TI - Direct evidence for destruction of polychlorobiphenyls by OH radicals in the subtropical troposphere. AB - Although laboratory studies have indicated that OH radicals readily oxidize polychorobiphenyls (PCBs) in the gas-phase, this mechanism has never been positively demonstrated under real atmospheric conditions. By applying elaborated sampling and analytical techniques we achieved for the first time simultaneously a field study of the diurnal atmospheric cycle of OH radical and PCBs in a remote site of eastern Mediterranean. In all cases, the concentration of sigmaPCB (sum of 27 congeners) showed a characteristic depletion during daytime, while the concentration of OH radicals was at the maximum levels. By assuming that the depletion of PCBs was caused solely by the destruction from OH radicals, PCB-OH rate constants (KOH) of different PCB congeners were determined from field data by applying the relative rate method. Our field-determined KOH values were notably coherent with those previously measured in laboratory experiments. In all measurement periods, the KOH values consistently decreased in sequence for those compounds showing an increasing degree of chlorination on the biphenyl group. By taking into account KOH values and latitude-dependent concentration of OH radicals, it was estimated that, near to tropical and subtropical regions, the atmospheric lifetimes of PCBs 8 and 110 should be substantially low (10 and 20 days, respectively). A significant fraction of PCBs should be destroyed during their residence over tropical/subtropical regions, due to the intensive destruction by OH. PMID- 12630471 TI - Laboratory studies of potential mechanisms of renoxification of tropospheric nitric acid. AB - Laboratory studies of the heterogeneous reactions between HNO3 in thin water films on silica surfaces and gaseous NO, CO, CH4, and SO2, proposed as potential "renoxification" mechanisms in the atmosphere, are reported. Transmission FTIR was used to monitor reactants and products on the silica surface and in the gas phase as a function of time. No reaction of CO, CH4, or SO2 was observed; upper limits to the reaction probabilities (gamma(rxn)) are < or = 10(-10) for CO and SO2 and < or = 10(-12) for CH4. However, the reaction of HNO3 with NO does occur with a lower limit for the reaction probability of gammaNO > or = (6 +/- 2) x 10( 9) (2s). The experimental evidence shows that the chemistry is insensitive to whether the substrate is pure silica or borosilicate glass. Nitric acid in its molecular form, and not the nitrate anion form, was shown to be the reactive species, and NH4NO3 was shown not to react with NO. The HNO3-NO reaction could be a significant means of renoxification of nitric acid on the surfaces of buildings and soils in the boundary layer of polluted urban atmospheres. This chemistry may help to resolve some discrepancies between model-predicted ozone and field observations in polluted urban atmospheres. PMID- 12630472 TI - Fractionation of platinum group elements in aqueous systems: comparative kinetics of palladium and platinum removal from seawater by Ulva lactuca L. AB - A marine macroalga, Ulva lactuca L., was used as a substrate to compare the kinetics of palladium (Pd) and platinum (Pt) removal from seawater. This work indicates that, while the equilibrium behaviors of Pd and Pt are in many respects similar, their kinetic behaviors are quite distinct. The removal of both Pt(II) and Pt(IV) from seawater by U. lactuca is slower than the removal of Pd(II) by approximately an order of magnitude. Relative Pd and Pt removal rates are strongly influenced by system hydrodynamics. Under quiescent conditions, lambda(Pd)/lambda(Pt), the ratio of Pd and Pt removal rates, is 7 +/- 2, whereas under turbulent conditions lambda(Pd)/lambda(Pt) can be as large as 27. These observations suggest that the disparate kinetic behaviors of Pd and Pt may produce considerable differences in the environmental dispersion of these elements. PMID- 12630473 TI - Kinetic modeling of TiO2-catalyzed photodegradation of trace levels of microcystin-LR. AB - A kinetic model has been developed to investigate the relative importance of major pathways for the photocatalytic degradation of trace levels of the cyanobacterial toxin microcystin-LR (MLR) in solutions containing a complex suite of dissolved organic matter and to test the sensitivity of MLR degradation to rate constants of the key processes. The kinetic model incorporates adsorption of the trace contaminant, other organics and oxygen on the particle surface, surface reactions between adsorbed radical and nonradical species, desorption of surface radical species, solution phase radical reactions, and radical termination pathways. Under conditions where the contaminant adsorbs strongly to semiconductor surface sites, rapid degradation is observed, and a primary degradation step appears to involve reaction between surface-located long-lived organic radicals (formed from hydroxyl radical scavenging by the bulk organic) and adsorbed trace contaminant. MLR degradation is relatively insensitive to changes in light intensity under these strongly adsorbing conditions but highly dependent under weakly adsorbing conditions and when solution phase degradation is important. While not verified independently, desorption of surface bound superoxide appears to lead to the production of organic peroxyl radicals through reaction of superoxide with the bulk organic. These solution phase organic peroxyl radicals are highly reactive and appear to be the primary source of trace contaminant degradation under conditions where the trace contaminant shows no observable adsorption and surface degradation is negligible. Under alkaline conditions, adsorption of carbonate onto the particle surface results in scavenging of surface hydroxyl radicals to form surface carbonate radicals that rapidly quench surface bound superoxide. This prevents organic peroxyl production, the primary agent of solution-phase trace contaminant degradation. PMID- 12630474 TI - Inhalation exposure to haloacetic acids and haloketones during showering. AB - Inhalation exposure to haloacetic acids (HAAs) and haloketones (HKs) in contaminated drinking water occurs during showering. The size distribution of the aerosols generated by a shower was determined using an eight size-range particle counter, which measured particles from 0.1 to >2 microm. An exponential increase in aerosol numbers was observed while the shower water was on, while the aerosol numbers declined exponentially once the water was turned off. The half-lives of the shower aerosols were longer than 5 min after the shower water was turned off. Although the majority of the shower-generated aerosols were smaller than 0.3 microm, these aerosols only contributed approximately 2% to the measured total aerosol mass. The total shower-generated particulate HAA and HK concentrations collected on an open face filter were approximately 6.3 and 0.13 microg/m3, respectively, for shower water HAA and HK concentrations of 250 and 25 microg/L, respectively. The vapor-phase HK concentrations were 25-50 microg/m3. The estimate of the dose from inhalation exposure of disinfection byproducts (DBPs) in the particulate phase indicate that they represent less than 1% of the ingestion dose, so inhalation is not expected to be an important exposure route to nonvolatile water contaminants or the portion of volatile DBPs that stay in the particulate phase, unless the lung is the target organ. The vapor-phase levels of volatile HKs, though, are significantly higher and can contribute greater than 10% of the ingestion dose during a shower. Thus, risk assessment to the these DBPs needs to consider the inhalation route. PMID- 12630475 TI - Relationship between particle mass and mobility for diesel exhaust particles. AB - We used the aerosol particle mass analyzer (APM) to measure the mass of mobility classified diesel exhaust particles. This information enabled us to determine the effective density and fractal dimension of diesel particles as a function of engine load. We found that the effective density decreases as particle size increases. TEM images showed that this occurs because particles become more highly agglomerated as size increases. Effective density and fractal dimension increased somewhat as engine load decreased. TEM images suggest that this occurs because these particles contain more condensed fuel and/or lubricating oil. Also, we observed higher effective densities when high-sulfur EPA fuel (approximately 360 ppm S) was used than for Fischer-Tropsch fuel (approximately 0 ppm S). In addition, the effective density provides the relationship between mobility and aerodynamic equivalent diameters. The relationship between these diameters enables us to intercompare, in terms of a common measure of size, mass distributions measured with the scanning mobility particle sizer (SMPS) and a MOUDI impactor without making any assumptions about particle shape or density. We show that mass distributions of diesel particles measured with the SMPS-APM are in good agreement with distributions measured with a MOUDI and a nano-MOUDI for particles larger than approximately 60 nm. However, significantly more mass and greater variation were observed by the nano-MOUDI for particles smaller than 40 nm than by the SMPS-APM. PMID- 12630476 TI - Influence of wettability and saturation on liquid-liquid interfacial area in porous media. AB - The knowledge of the area of interfaces between phases is important to understand and quantify many flow and transport processes in porous media. In this work, we apply the interfacial tracer technique to study the dependence of fluid-fluid interfacial area on saturation and wettability. The interfacial area between the wetting and the nonwetting phases (brine and decane) in unconsolidated porous media (glass beads) was measured using an anionic surfactant (3-phenyl decyl benzene sulfonate) as an interfacial tracer. The beads are water-wet; treating them with organosilane rendered them oil-wet. The measurements were done at a series of steady-state fractional flows, providing data at intermediate as well as residual saturations. Flow rates were kept low so that capillary forces controlled the fluid configurations. We observe significant differences in interfacial areas as a function of wetting-phase saturation as the wettability is changed from water-wet to oil-wet. During primary drainage, measured interfacial area increases monotonically with decreasing water saturation in a water-wet medium. In contrast, the interfacial area measured in the oil-wet porous medium increases with decreasing decane saturation, reaches a maximum, and decreases as the residual decane saturation is achieved. The oil-wet experiment is qualitatively consistent with theoretical results that predict the existence of a maximum in fluid-fluid interfacial area during drainage. The water-wet experiment is consistent with theoretical predictions that include the area of grains in pores that have been drained. We conclude that, in the water-wet experiments, the tracer adsorbs at the interface between the nonwetting phase and the wetting films on grains. In the oil-wet experiments, either the oil films are not sustained at high water saturation or the tracer does not adsorb at them, possibly prevented by steric hindrance. Interpretation of interfacial tracer experiments therefore requires care: for some mass transport processes, the thin films of wetting phase on grains will not behave the same as macroscopic volumes of wetting phase. PMID- 12630477 TI - Method comparison study for weak acid dissociation cyanide analysis. AB - Method comparison studies of two different methods for the analysis of weak acid dissociable (WAD) cyanide revealed analytical flaws and/or matrix interference problems with both procedures. EPA "draft" method 1677 using a Perstorp 3202 CN analyzer was compared to Standard Method 4500 CN I. It was discovered that the Perstorp analyzer produced more precise and more accurate results once appropriate and necessary procedural steps from the EPA draft method were modified. Comparison of these two methods, was based on "real world" samples collected from a mine-tailing solution. The mine-tailing solution contained high concentrations of cyanide and metals. Inconsistencies in method procedures were traced to sulfide interferences and high concentrations of WAD metals. Conclusions were based upon a large sample base collected from a mine site over a 90-day period. PMID- 12630478 TI - Recovery of semivolatile organic compounds during sample preparation: implications for characterization of airborne particulate matter. AB - Semivolatile compounds present special analytical challenges not met by conventional methods for analysis of ambient particulate matter (PM). Accurate quantification of PM-associated organic compounds requires validation of the laboratory procedures for recovery over a wide volatility and polarity range. To meet these challenges, solutions of n-alkanes (nC12-nC40) and polycyclic aromatic hydrocarbons PAHs (naphthalene to benzo[ghi]perylene) were reduced in volume from a solvent mixture (equal volumes of hexane, dichloromethane and methanol), to examine recovery after reduction in volume. When the extract solution volume reached 0.5 mL the solvent was entirely methanol, and the recovery averaged 60% for n-alkanes nC12-nC25 and PAHs from naphthalene to chrysene. Recovery of higher MW compounds decreased with MW, because of their insolubility in methanol. When the walls of the flasks were washed with 1 mL of equal parts hexane and dichloromethane (to reconstruct the original solvent composition), the recovery of nC18 and higher MW compounds increased dramatically, up to 100% for nC22-nC32 and then slowly decreasing with MW due to insolubility. To examine recovery during extraction of the components of the High Capacity Integrated Gas and Particle Sampler, the same standards were used to spike its denuders and filters. For XAD-4 coated denuders and filters, normalized recovery was >95% after two extractions. Recovery from spiked quartz filters matched the recovery from the coated surfaces for alkanes nC18 and larger and for fluoranthene and larger PAHs. Lower MW compounds evaporated from the quartz filter with the spiking solvent. This careful approach allowed quantification of organics by correcting for volatility- and solubility-related sample preparation losses. This method is illustrated for an ambient sample collected with this sampler during the Texas Air Quality Study 2000. PMID- 12630479 TI - Selenium removal by constructed wetlands: quantitative importance of biological volatilization in the treatment of selenium-laden agricultural drainage water. AB - Management of selenium (Se) -contaminated agricultural drainage water is one of the most important environmental issues in California. To evaluate the feasibility of utilizing constructed wetlands to remediate Se-laden drainage water and the role of biological volatilization in Se removal, 10 flow-through wetland cells were constructed in 1996 in Corcoran, California. The monthly monitoring study from May 1997 to December 1999 showed that the vegetated wetlands were capable of significantly reducing Se from the inflow drainage water; an average of 69.2% of the total Se mass in the inflow was removed. Most of the Se was retained in sediment, and <5% of the Se was accumulated in plant tissues. Selenium volatilization was highest in the rabbitfoot grass wetland cell, where 9.4% of the Se input was volatilized over a 2-year period. Volatilization was greater in spring and summer than in fall and winter. For example, in May and June of 1998, 35 and 48%, respectively, of the Se entering the rabbitfoot grass cell was volatilized, whereas in the winter months, <5% was volatilized. The feasibility of using constructed wetlands for Se remediation, methods for the enhancement of Se volatilization, and the importance of considering potential Se ecotoxicity are discussed. PMID- 12630480 TI - Interpreting deposition patterns of microbial particles in laboratory-scale column experiments. AB - The transport and fate of microbial particles in subsurface environments is controlled by their capture (natural filtration) by sediment grains. Typically, filtration models used to describe microbe removal in porous media predict exponential decrease in microbial particle concentration with travel distance. However, a growing body of laboratory-scale column experiments suggests that the retained microbial particle profiles decay nonexponentially. The observed behavior may be attributed to the heterogeneity in the interactions between microbial particles and sediment grains, most likely due to the inherent variability in the microbial particles. This factor can be incorporated into classical colloid filtration (deposition) theory by inclusion of a distribution in the deposition rate coefficient. We show that certain distributions of the deposition rate coefficient (i.e., log-normal, bimodal, and power-law distributions) give rise to nonexponential deposition patterns. Comparisons of model predictions to experimental data indicate that the observed nonexponential deposition behavior of bacteria and virus particles may be attributed to a broad range (i.e., a power-law distribution) of microbial deposition rates. Other mechanisms such as particle release and blocking by previously deposited microbial particles are also shown to be potential sources of deviation from the classical filtration theory. Our results further suggest that monitoring fluid phase particle concentration is insufficient for accurate characterization of the deposition and transport behavior of microbial particles in saturated porous media. Rather, the shape of the microbial particle retention profile is shown to be a key indicator of the mechanisms controlling microbial deposition and transport. PMID- 12630481 TI - Induced phytoextraction/soil washing of lead using biodegradable chelate and permeable barriers. AB - Chelate-induced remediation has been proposed as an effective tool for the extraction of lead (Pb) from contaminated soils by plants. However, side-effects, mainly mobilization and leaching of Pb, raise environmental concerns. Biodegradable, synthetic organic chelate ethylenediaminedisuccinic acid (EDDS), and commonly used ethylenedimanetetraacetic acid (EDTA) were used for induced phytoextraction with a test plant Brassica rapa and in situ washing of soil contaminated with 1350 mg/kg of Pb. Horizontal permeable barriers were placed 20 cm deep in soil columns and tested for their ability to prevent leaching of Pb. The reactive materials in the barriers were nutrient enriched vermiculite, peat or agricultural hydrogel, and apatite. EDTA and EDDS addition increased Pb concentrations in the test plant by 158 and 89 times compared to the control, to 817 and 464 mg/kg, respectively. In EDTA treatments, approximately 25% or more of total initial soil Pb was leached in single cycle of chelate addition. In EDDS treatments, 20% of the initial Pb was leached from columns with no barrier, while barriers with vermiculite or hydrogel and apatite decreased leaching by more than 60 times, to 0.35%. 11.6% of total initial Pb was washed from the soil above the barrier with vermiculite and apatite, where almost all leached Pb was accumulated. Results indicate that use of biodegradable chelate EDDS and permeable barriers may lead to environmentally safe induced Pb phytoextraction and in situ washing of Pb. PMID- 12630482 TI - Quantifying the emission benefits of opacity testing and repair of heavy-duty diesel vehicles. AB - The objective of this study was to begin to quantify the benefits of a smoke opacity-based (SAE J1667 test) inspection and maintenance program. Twenty-six vehicles exhibiting visible smoke emissions were recruited: 14 pre-1991 vehicles and 12 1991 and later model year vehicles. Smoke opacity and regulated pollutant emissions via chassis dynamometer were measured, with testing conducted at 1609 m above sea level. Twenty of the vehicles were then repaired with the goal of lowering visible smoke emission, and the smoke opacity testing and pollutant emissions measurements were repeated. For the pre-1991 vehicles actually repaired, pre-repair smoke opacity averaged 39% and PM averaged 5.6 g/mi. NOx emissions averaged 22.1 g/mi. After repair, the average smoke opacity had declined to 26% and PM declined to 3.3 g/mi, while NOx emissions increased to 30.9 g/mi. For the 1991 and newer vehicles repaired, pre-repair smoke opacity averaged 59% and PM averaged 2.2 g/mi. NOx emissions averaged 12.1 g/mi. After repair, the average opacity had declined to 30% and PM declined to 1.3 g/mi, while NOx increased slightly to 14.4 g/mi. For vehicles failing the California opacity test at >55% for pre-1991 and >40% for 1991 and later model years, the changes in emissions exhibited a high degree of statistical significance. The average cost of repairs was 1088 dollars, and the average is very similar for both the pre-1991 and 1991+ model year groups. Smoke opacity was shown to be a relatively poor predictor of driving cycle PM emissions. Peak CO or peak CO and THC as measured during a snap-acceleration were much better predictors of driving cycle PM emissions. PMID- 12630483 TI - Allometric modeling of plant root growth and its application in rhizosphere remediation of soil contaminants. AB - Allometric curves relating tree trunk diameter to root biomass, depth, and breadth were compiled for mulberry (Morus sp.). The curves were based on statistical analyses of measurements made on 29 different-sized trees ranging in age from 2 to 15 yr that had grown from seed in a naturally revegetated former sludge basin containing polyaromatic hydrocarbons. Over a 15-yr period, the curves indicate that the fine root biomass (<1.5 mm diameter) increases 60-fold and, under the right circumstances, can be a part of a root system that reaches a 2-m depth. The fine roots of mulberry were shown to produce several flavonoid compounds at concentrations (ranging from 94 to 525 microg/cm3) known to support the growth of organisms capable of degrading xenobiotics. Recognizing the root system as the driver of rhizoremediation, allometry curves presented in this paper can be used to quantify the magnitude of the driver (root system) without damaging plants during the course of a multiyear field study. PMID- 12630484 TI - Fe(O)/lignitic coal: an efficient and mechanically stable reactive material for purification of water containing heavy metals, radionuclides, and nitroaromatics. AB - Mine water in the former uranium mine of Konigstein (Saxony, Germany) contains high concentrations of acid, sulfate, iron, aluminum, various heavy metals, radionuclides, and nitroaromatics. Research has been conducted for several years to establish the extent to which reduction of pollutant concentrations can be positively influenced and accelerated by storage of reactive materials in mine cavities. Investigations were made at different scales to test and select materials with respect to maximum fixation of contaminants (underground column tests) to examine hydraulic effects (underground large-scale column tests) and to optimize material properties (laboratory tests). The investigations have shown that a mixture of Fe chips and lignitic coal is capable of efficiently cleaning acid and contaminant-containing mine water. The examined material is easily available and compatible with the environment. A large-scale application of such a reactive barrier is being considered for mine water treatment in the future and is deemed to be a reasonable conception for a safety component after conclusion of the flooding. PMID- 12630485 TI - Comparison of the recyclability of flame-retarded plastics. AB - Mechanical recycling of plastics from waste from electrical and electronical equipment (WEEE) is increasingly expected by regulators and demanded by original equipment manufacturers (CEMs); however, mechanical recycling is generally recognized to be the most economically costly and technically challenging method of recovering WEEE plastics. With 12% of WEEE plastics requiring the use of flame retardants in order to ensure appropriate levels of consumer fire safety, there is a distinct need for data from comparative tests on recyclability of various flame-retarded plastics. Ten commercially available flame-retarded plastic grades commonly used in electronic equipment (eight "halogen-free" grades and two grades containing brominated flame-retardants (BFRs)) were subjected to two different recycling scenarios. A standard recycling scenario was carried out by repeatedly extruding the materials and an accelerated hydrolysis scenario was carried out to study the influence of humidity from air during use on the process. Both, virgin and recycled materials were tested for a potential formation of polybrominated dibenzodioxins/furans (PBDD/Fs), their mechanical properties were assessed and the fire safety rating was determined. Results indicate that none of the tested materials showed a potential to form the PBDD/Fs regulated by the German Chemicals Banning Ordinance. The halogen-free plastic grades showed a significant deterioration of mechanical properties after recycling, whereas those plastics containing BFRs were able to pass all test criteria, thus maintaining their original properties. With respect to the fire safety rating, none of the eight tested halogen-free plastic grades could maintain their fire safety rating after five recycling loops, whereas both BFR plastics continued to achieve their fire safety ratings. Therefore the tested BFR containing plastic materials showed superior recycling properties compared to the tested halogen-free plastic grades with respect to all investigated parameters. PMID- 12630486 TI - Comment on "Trace metal levels in uncontaminated groundwater of a coastal watershed: importance of colloidal forms". PMID- 12630487 TI - Curtailing maternal to child transmission of HIV. PMID- 12630489 TI - Minimising antibiotic resistance to Staphylococcus aureus in developing countries. AB - OBJECTIVE: To assess the role of rational drug use and laboratory service in preventing the emergence of multiple antibiotic resistant Staphylococcus aureus in developing countries. DATA SOURCE: Literature search on compact disk-read only memory (CD-ROM) Medline and Internet using the key words: Staphylococcus and antibiotic resistance. A few articles were manually reviewed. STUDY SELECTION: Relevant studies or articles on antibiotic resistance with special reference to Eastern Africa, region are included in the review. DATA EXTRACTION: From individual studies or articles. DATA SYNTHESIS: Evidence for the spread of S. aureus multiple antibiotic resistance is synchronized under the headings: Introduction, current situation, antibiotic resistance control strategies, are outlined. CONCLUSION: There is need for concerted efforts between different groups to monitor changes in the epidemiology and antibiotic resistance of S. aureus. Strategies aimed at preventing transmission of resistant strains are remarkably effective when strictly enforced. Necessary attention should be given on the subject so that meaningful control measures preventing the expansion of antimicrobial resistance can be formulated, thereby ensuring the future successful treatment of Staphylococcal infections. PMID- 12630488 TI - Maternal immune responses and risk of infant infection with HIV-1 after a short course Zidovudine in a cohort of HIV-1 infected pregnant women in rural Kenya. AB - OBJECTIVE: To investigate the effects of short-course nucleoside reverse transcriptase inhibitor (Zidovudine, ZDW/AZT) on maternal immune responses and risk of infant infection with HIV-1 among rural-based mothers in western Kenya. DESIGN: A prospective cohort study involving HIV-1 seropositive pregnant mothers and their infants. SUBJECTS: One hundred and seven HIV-1 seropositive asymptomatic pregnant women and their infants. METHODS: After informed consent, the women were enrolled at gestation age between 16-24 weeks. For cultural and economic reasons, all mothers were allowed to breast feed their infants. Short course antepartum regime of AZT was administered to all mothers starting at 36 weeks gestation until start of labour. Maternal absolute CD4+ T cell subset assays were performed before 3rd trimester (about 36 weeks gestation) and after a 4-week therapy of AZT (at least one month post-nuptially). Infant HIV-1 status was determined by HIV-1 DNA polymerase chain reaction (PCR) on samples sequentially taken at 1, 2, 3, 4, 6 and 9 months and confirmed by serology at 18 months of age. INTERVENTIONS: Antepartum short-course orally administered AZT: 300mg twice-daily starting at 36 weeks gestation until start of labour, 300mg at labour onset and 300mg every three hours during labour until delivery. MAIN OUTCOME MEASURES: Maternal CD4+ T cell counts before and after AZT treatment. Determination of infant HIV-1 infection status. RESULTS: Among 107 women sampled, only 59 received full dose of AZT and thus qualified for present analysis. Of these, 12 infected their children with HIV, while 47 did not. Comparison of CD4+ T cells before and after AZT treatment scored a significant rise in all mothers (P = 0.01). This increase in CD4+ T cells was not significant among mothers who infected their infants with HIV-1 (P = 0.474). However, a significant rise in CD4+ T cells following AZT therapy was observed only in mothers who did not transmit HIV-1 to their infants (P=0.014). CONCLUSION: These data suggest that a rise in the CD4+ T cell counts following short AZT regimen, now widely in use in resource-weak countries, may be evidence of the active suppression of the replication of HIV. However, further studies to examine the multi-factorial effect of CD4+ lymphocytes and pregnancy on MTCT of HIV need to be carried out to help fully explain the effect of AZT on immune response and whether the CD4+T cell count can be used as a true test of immunological normalisation during antiretroviral therapy. PMID- 12630490 TI - Management of tracheo-bronchial foreign bodies in children. AB - OBJECTIVES: To review alternative surgical and anaesthetic options in the management of foreign bodies lodged in the tracheobronchial tree in children aged below 10 years. DESIGN: A five year retrospective secondary data analysis. SETTING: Three hospitals based in Eldoret Municipality, Kenya. MAIN OUTCOME MEASURES: Outcome variables included morbidity and mortality. RESULTS: Of the thirty two children studied, 74.1% had bronchoscopy and 25.9% thoracotomy. The overall mortality rate was 3.3%. CONCLUSION: Foreign bodies in the tracheo bronchial tree are a major cause of morbidity and mortality in children aged below 10 years. In settings where appropriate equipment is inadequate, timely thoracotomy may be life saving. PMID- 12630491 TI - Management of solitary thyroid nodules in rural Africa. AB - OBJECTIVES: To review a simple protocol for the management of solitary thyroid nodules and to document the spectrum of pathological diagnoses associated with this condition. DESIGN: A retrospective review of all solitary solid thyroid nodules excised over a three-year period from 1st January 1999 to 31st December 2001. SETTING: A rural church-based hospital in Kenya. SUBJECTS: All patients undergoing thyroidectomy for solitary solid thyroid nodule over a three-year period at Kijabe Hospital. INTERVENTIONS: A simple protocol was used to manage this condition involving history, clinical examination, needle aspiration of the lesion, and excision when clinically indicated. MAIN OUTCOME MEASURES: Clinical diagnosis, tribe, operation performed, pathology, and complications of surgery. RESULTS: Eighty-one operations were performed for a solitary thyroid nodule. The most common operations were lobectomy and isthmusectomy. There were two complications--a neck haematoma that required surgery and one recurrent laryngeal nerve injury. The commonest pathological diagnosis was multinodular goitre (42%). There was a 16% malignancy rate with eight papillary carcinomas, five follicular carcinomas, and one hurthle cell carcinoma. CONCLUSIONS: The simple protocol described gives good results in a rural African hospital. Solitary solid thyroid nodules should be routinely excised due to the 16% malignancy rate in this condition. There is a possibility that there is a shift in the ratio of papillary to follicular carcinomas compared to older African studies and this would be an interesting area for further study. PMID- 12630492 TI - Antibacterial effect of Zingiber officinale and Garcinia kola on respiratory tract pathogens. AB - OBJECTIVE: To investigate the antibacterial activity of Zingiber officinale (ginger) Garcinia kola (bitter kola) on four respiratory tract pathogens. DESIGN: A prospective study based on laboratory investigations. SETTING: Department of Life Sciences, University of Buea. Throat swabs were collected from 333 individuals with running nostrils, cough and/or catarrh in three localities of Buea namely Bokwango, Molyko and Bolifamba. Staphylococcus aureus, Streptococcus pyogenes, Streptococcus pneumoniae and Haemophilus influenzae were isolated from the specimens using standard microbiological procedures. The antibacterial activity of ethanolic extracts of ginger and bitter kola, were investigated on these pathogens using the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays. RESULTS: The extracts exhibited antibacterial activity against the pathogens. The MIC of extracts ranged from 0.0003 microg/ml to 0.7 microg/ml for ginger and 0.00008 microg/ml, to 1.8 microg/mL for bitter kola, while MBC ranged from 0.1.35 microg/ml to 2.04 microg/ml for ginger and 0.135 microg/ml to 4.2 microg/ml for bitter kola. CONCLUSION: Results indicated that extracts of ginger root and bitter kola may contain compounds with therapeutic activity. PMID- 12630493 TI - Chronic myeloid leukaemia at the Kenyatta National Hospital, Nairobi. AB - OBJECTIVE: To determine the clinical and haematological factors associated with treatment and outcome of chronic myeloid leukaemia (CML) at Kenyatta National Hospital. DESIGN: Retrospective survey of patients treated for chronic myeloid leukaemia. SETTING: Kenyatta National hospital, Nairobi, Kenya, between April 1990 and August 2000. SUBJECTS: Patients with chronic myeloid leukaemia. RESULTS: One hundred and four patients, 55 males and 49 females, age range 10-72 years with a median age of 35 years. Treatment with busulphan getting less popular in favour of hydroxyurea. Median follow-up 20 months with none of the clinical and haematological parameters impacting significantly on duration of follow-up. CONCLUSION: CML occurs at a younger age-group in Kenya, and none of the clinical or haematological parameters appears to impact on follow-up duration. PMID- 12630494 TI - Effect of micronutrient fortified beverage on nutritional anaemia during pregnancy. AB - OBJECTIVE: To evaluate the efficacy of a multiple micronutrient fortified beverage containing eleven nutrients at physiological levels in prevention of anaemia and improving iron and vitamin A status during pregnancy. DESIGN: A randomised double blind placebo controlled study. SETTING: Mpwapwa and Kongwa Districts in Dodoma Region of Tanzania. SUBJECTS: Five hundred and seventy nine pregnant women were screened for entry into the study and 439 women who met the study criteria were enrolled. INTERVENTIONS: Study participants received either a fortified (F) or non-fortified (NF) orange flavoured drinks identical in appearance, provided in two self administered servings per day for an eight week period. MAIN OUTCOME MEASURES: Comparison of haemoglobin (Hb), serum ferritin (SF) and serum retinol (SR) at baseline and follow up. RESULTS: After eight weeks of supplementation, the F group (n=129) had a significantly higher Hb increase of 0.86 g/dL compared to 0.45 g/dL in the NF group (n=130) p<0.0001. Gestational age at entry into the study, moderated the effect on Hb of the fortified drink. Women at earlier gestational age upon entry, had a higher rise in Hb than women of late gestational age (0.8 g/dL versus 0.04 g/dL rise respectively, p=0.038, n=188). The risk of being anaemic at the end of the study for those in the F group was reduced by 51% (RR=0.49, CI=0.28 to 0.85). Iron stores (by serum ferritin levels) increased by 3 microg/L in the F group (p=0.012) and a decrease of 2 microg/L in the NF group (p=0.115). The follow up ferritin concentration depended on initial ferritin level. Regardless of treatment group, serum retinol concentrations were significantly higher in mothers who had delivered. Mothers who had adequate levels at entry benefited more from the supplement than those with low levels (0.26 micromol/L versus no significant difference). CONCLUSIONS: The multiple micronutrient-fortified beverage given for eight weeks to pregnant women improved their haemoglobin, serum ferritin and retinol status. The risk for anaemia was also significantly reduced. The important predictors of Hb increase at follow up were the fortified beverage, baseline Hb, serum retinol, baseline ferritin and gestational age at entry into study. Anthropological research showed that the beverage was highly acceptable and well liked. PMID- 12630495 TI - Pathogenic intestinal parasites and bacterial agents in solid wastes. AB - OBJECTIVE: To determine the profile of potentially pathogenic enteric parasites and bacterial agents in municipal refuse dumps in Ibadan, Nigeria. DESIGN: A cross-sectional survey. SETTING: Five major market places refuse dumps in Ibadan municipality, Nigeria. METHODS: The major market places in Ibadan city were randomly selected by lottery method. The refuse sludge were sampled and examined parasitologically and bacteriologically using the methods as described. Data analysis was done by using chi-square test where applicable. RESULTS: Cases of multiple intestinal parasites and bacterial agents were commonly encountered in the sludge refuse samples. The commonly found parasitic agents were of both human and veterinary importance. These include Ascaris Lumbricoides (9.3 egp), Entamoeba histolytica (8.07 cyst per gram), Hookworm/strongyle (6.27 egp) and Ascaris suum (1.07 egp). Others were Ascaris vitolorum (1.09 epg), Strongyloides papillosu (0.52 larvae per gram.), Schistosoma suis (0.31 epg) Dicrocoelium dendriticum (0.9 egp). The most commonly found bacterial agents were Klebsiella species, Escherichia Coli, Proteus species, streptococci, and other gram-positive organisms. Climatic conditions affected the distribution of both parasites and bacterial agents in the sludge (P<0.001). More intestinal parasites 2423 (53.4%) and bacterial agents 2150 (27.2%) were encountered at mean air temperature 26.1 +/- 0.6, mean relative humidity of 72 +/- 3.5%. The degree of contamination by market locations varies significantly (P<0.001). CONCLUSION: A high degree of contamination of solid waste dumpsites with bacterial and parasitic agents was observed in the present study. As a result of the public health importance of the organisms isolated it is opined that well planned waste management and health education programs will go a long way to reduce the potential epidemic risks posed by such sites in Ibadan, Nigeria. PMID- 12630497 TI - Neurofibromatosis type 1: report of two contrasting cases. AB - We present two cases of neurofibromatosis type 1 (NF-1), one a 35 year old male who first recognised his problem at the age of fifteen years and at the time of assessment, satisfied the National Institute of Health (NIH) diagnostic criteria for NF-1 and had a nodular plexiform neurofibroma involving the left fifth dorsal nerve root and a diffuse plexiform neurofibroma involving the left lower limb. The second patient, a 45 year old female recognised her problem at the age of 39 years, did not quite satisfy the NIH diagnostic criteria for NF 1 and had diffuse plexiform neurofibroma involving both lower limbs and buttocks almost symmetrically, a finding which has not previously been described to the best of our knowledge. The scarcity of management options are briefly outlined. PMID- 12630496 TI - Ureteric injuries following pelvic operations. AB - BACKGROUND: Iatrogenic injuries to the ureter are hazardous complications of pelvic operations, causing severe morbidity and even mortality. OBJECTIVE: To present our 10 years experience in the management of such ureteric injuries. DESIGN: A retrospective study carried out between January 1990 and December 1999. SETTING: Two busy health institutions, namely Ife State hospital and Wesley Guild Hospital, both of the Obafemi Awolowo University (OAU) Teaching Hospitals Complex, Ile-Ife, Nigeria. RESULTS: The incidence of iatrogenic injury was 0.4%. Ureteral transection was the commonest lesion (58%). Ureteroneocystostomy was performed in 70% of the operated cases. Those diagnosed at the time of injury and treated with end-to-end anastomosis had the best results. CONCLUSION: The proper identification and, when necessary, isolation of the ureter during operations in which there is a risk is crucial in reducing the incidence of ureteral injuries. PMID- 12630498 TI - Rubella seroprevalence in adolescent girls in the eastern region of Turkey. PMID- 12630499 TI - The "business" of fat: an economic point of view. PMID- 12630501 TI - An activity calendar program for children who are overweight. AB - Sedentary lifestyle activities, such as computer use and television viewing, are modifiable causes of overweight among children. There are many recommendations in the literature that suggest the number of children who are overweight must be decreased; however, none of the research describes an instrument to achieve this goal. This article describes the use of a home-based physical activity calendar and its successes and failures. PMID- 12630500 TI - A primer on early childhood obesity and parental influence. AB - Childhood obesity is a common health problem facing U.S. children with an increasing prevalence particularly in certain populations. Recognizing obesity in children is a clinical determination with specific measures that can indicate potential future associated health problems, but parental perception of overweight and/or obesity is influenced by other conditions. This primer offers a brief synopsis of parental influence in the etiology of early childhood obesity beginning with parameters of obesity and how it is operationalized through measurement. The importance of parental perceptions of their children relative to obesity and eating are discussed, and how parents influence the development of childhood eating behaviors or physical activity are considered. PMID- 12630502 TI - Creating Healthy Futures: an innovative nurse-managed transition clinic for adolescents and young adults with special health care needs. AB - The Creating Healthy Futures (CHF) clinic is an innovative, nurse-managed service model that provides comprehensive transition services to youth and young adults with special health care needs. Unlike other transition service models reported in the literature that are medically based, comprehensive transition services were coordinated by a family nurse practitioner who worked in close collaboration with an interagency team of pediatric and adult service providers. The Creating Healthy Futures (CHF) service model was originally developed for use in educational settings but was adapted for use in health care settings. The components of the service model are described in detail as a model for replication in other health care settings. PMID- 12630503 TI - Fever management audit: Australian nurses' antipyretic usage. AB - Do nurses manage fevers of children hospitalized for a febrile illness ritualistically or rationally? Nurses recorded temperatures more frequently during the first 8 hours in the ward with a mean frequency of 13.36 (SD = 4.76, range 5 to 24) during the first 24 hours following admission. In the majority of cases, there was a strong second hourly pattern of temperature monitoring according to the time of day (e.g., 0600 hr, 0800 hrs, 1000 hr). Seventy-six percent (51) of the children received at least one antipyretic. The mean temperature when antipyretics were administered was 38.34 degrees C (SD = 1.02, range 35.9 degrees C to 40.8 degrees C). The highest antipyretic administration occurred during the daytime, and the highest temperature recording occurred during the nighttime. Antipyretic administration and mean temperatures generally followed a similar pattern, except at 0800 and 1600 hours when antipyretic administration was high and mean temperatures low. This study revealed a need to further investigate the knowledge, attitudes, and decision-making criteria of nurses toward fever management. PMID- 12630504 TI - Pediatric management problems. Periorbital cellulitis. PMID- 12630505 TI - Foster child health and development: implications for primary care. AB - Children in foster care are the most vulnerable to experiencing poor health compared with any other group of children in the United States. Children enter foster care due to experiences that have been detrimental to their health and well-being, including child abuse and neglect. They have significantly higher rates of all health problems than the general population of children, including acute and chronic illnesses, growth and developmental problems, serious mental health problems, and difficulties accessing health services. Additionally, they often experience multiple foster care placement transitions, involvement in multiple systems of care (e.g., mental health, juvenile justice, special education), and aging-out of foster care at age 18 before they are developmentally ready for independent living. While management of the complex health and developmental needs of these children is challenging, nurses in primary care have the expertise and obligation to serve these vulnerable children. Priorities for clinical practice include care coordination and interdisciplinary collaboration; maintenance of adequate, up-to-date health records; vigorous follow-up; and health-targeted advocacy. PMID- 12630506 TI - Helping children through an adult's serious illness: roles of the pediatric nurse. AB - Of the many life crises that a child may encounter on the path to adulthood, one of the most common is the serious or terminal illness of a loved adult. It is a reality that facing such illness can result in trauma, insecurity, and future psychological and behavioral problems. Alternatively, such challenges can produce enhanced family communication, increased compassion, and coping skills that can last a lifetime. The pediatric nurse, by embracing core principles of openness and honesty and by providing concrete developmental information, can empower parents to support their own children. Nurses use family-centered assessment skills to determine the impact of an adult's illness on the child and then guide the parent through those topics that are most likely to be of concern, including (a) the name of the illness, (b) the cause of the illness, (c) the potential contagion or spread of the illness, and (d) the ultimate impact of the illness on the life of the child. PMID- 12630507 TI - Adolescent suicide attempt: undisclosed secrets. PMID- 12630508 TI - Examining the validity, reliability, and preference of three pediatric pain measurement tools in African-American children. AB - PURPOSE: To compare the validity, reliability, and preference of pain intensity measurement tools--the African-American Oucher Scale, the Wong-Baker FACES Scale, and the Visual Analog Scale (VAS)--in 100 African-American children between 3 and 18 years of age. METHODS: During a visit to the sickle cell anemia clinic, the children were asked to describe two painful procedures/treatments they had experienced and to choose which one hurt the most. They rated the pain intensity of each procedure/treatment on the three scales that were presented in a preselected order. After using all the scales, the children then chose their favorite scale. The same procedure was repeated at the end of the visit, with a minimum of 15 minutes between the test and retest. RESULTS: Concurrent validity was determined by comparing the rank order of the two identified painful items with the pain scores on the three scales. Chi square analysis indicated a statistically significant difference among the scales for preference (chi2 = 24.08, df = 2, p < 0.0001) and reliability (chi2 = 6.12, df = 2, p < 0.05), but not validity (chi2 = 0.81, df = 2, NS) for the entire sample. Most children preferred using the FACES Scale (56%) versus the Oucher (26%) and VAS (18%). When the data were analyzed for three cognitive stages (ages 3-7, 8-12, and 13-18 years), the following were found: For the school-age period, chi square for reliability did not reach significance (chi2 = 4.35, df = 2, NS); for the preschool-age period, chi square for validity did reach significance (chi2 = 6.49, df = 2, p < 0.05) and was strongest for the FACES scale, followed by the Oucher. CONCLUSIONS: The study findings indicated that the FACES and African American Oucher Scales are valid and reliable tools for measuring pain in children. Among the entire sample and each age group, the FACES scale was the most preferred scale. PMID- 12630509 TI - The effectiveness of anti-leukotriene agents in childhood asthma: evidence to guide clinical practice. PMID- 12630510 TI - Real stories: Aspiring to become a nurse. AB - With the current crisis in the health care workforce and the predictions of looming shortages yet to come, it is imperative that we begin to appreciate why pediatric nurses choose their careers. What life experiences contribute to our desire to join a "club" that can boast its members want to serve others? How does a nursing career look to a high school senior or an outsider who is contemplating embarking on the rigorous education that it takes and the long hours that are promised at the end? These stories give us insight into how nurses today and potential nurses in the future view their life's contribution to their career choice. PMID- 12630511 TI - Ethical principles and parental choice: treatment options for neonates with hypoplastic left heart syndrome. AB - Nurses caring for children with congenital heart disease face unique challenges, especially when caring for neonates diagnosed with hypoplastic left heart syndrome (HLHS). The treatment options for these neonates present difficult choices for the child's decision makers and are not without significant life altering consequences. In order to assist in the decision-making process, nurses as patient and family advocates should acknowledge the unique role they play in the informed consent process, while simultaneously identifying specific ethical principles that are components of this process. By incorporating the principles of autonomy, beneficence, and veracity into specific nursing interventions, nurses can assist families in making informed decisions regarding a treatment option that is best for the child as well as the family. PMID- 12630512 TI - Messy conversations and the willingness to be disturbed. AB - Organizations operate successfully on how the parts interact together, not how they act independently. The only way to improve the world is through relationships, and conversations are the prelude to creating that change. Having the courage to reach out and start the dialogue knowing full well that it might be very messy is the only way to end the fragmentation and loss of community we experience. PMID- 12630513 TI - The growth of AIDS orphans and policy solutions. PMID- 12630514 TI - A new era of body decoration: what are kids doing to their bodies? PMID- 12630515 TI - Respite care and Caring Communitites, Inc. PMID- 12630516 TI - An awakened attitude toward improving bed rail safety. PMID- 12630517 TI - Serum S 100 B: a marker of brain damage in traumatic brain injury with and without multiple trauma. AB - This prospective clinical study was conducted to determine whether S 100 B is a reliable serum marker for traumatic brain injury (TBI) with and without multiple trauma. Fifty-five trauma patients (Injury Severity Score [ISS] > or = 24 and Glasgow Coma Score [GCS] < or = 8) were classified by radiography, computer tomography, ultrasound, and neurology as TBI without multiple trauma (n = 23), TBI with multiple trauma (n = 23), or multiple trauma without TBI (n = 9). S 100 B was measured initially after trauma and daily for a maximum of 21 days. Both survivors and nonsurvivors had markedly increased S 100 B initially. All survivors returned to normal or moderately increased S 100 B levels within the first 48 h after trauma. In contrast, all nonsurvivors of isolated TBI had S 100 B values that either increased consistently or dropped and then increased again 48 h after the initial increase after trauma. There was no relationship between localization, extent, or severity of TBI and S 100 B. According to receiver operating characteristic curve analysis and calculation of the area under the curve (AUC), S 100 B is equally accurate for mortality prediction at 24, 48, and 72 h after trauma and is most accurate >84 h after trauma. Sensitivity/specificity for mortality prediction are more accurate in TBI without multiple trauma (AUC 0.802-0.971) than in TBI with multiple trauma (AUC 0.693 0.783). Thus, though S 100 B may be a reliable marker of brain damage in TBI without multiple trauma 24 h after trauma and thereafter, it appears to be less reliable in TBI with multiple trauma. PMID- 12630518 TI - Fas-mediated neutrophil apoptosis and associated A1 protein expression during systemic inflammation are regulated independently of both tumor necrosis factor receptors. AB - TNFR-1 (p55) and Fas share a death domain which is critical for apoptosis signaling whereas TNFR-p55 and TNFR-2 (p75) can activate NF-kappaB leading to anti-apoptotic proteins expression such as A1. The purpose of this study was to elucidate the role(s) of TNFR-p55 and TNFR-p75 in Fas-mediated neutrophil apoptosis and A1 expression in a mouse model of endotoxemia. Gene knockout (KO) (p55-/-, p75-/-, p55(-/-)/p75(-/-)) or wild type (WT) mice were injected i.p. with saline or LPS (4 microg/g) followed by collecting peripheral blood after 24 h. Neutrophil apoptosis was assessed by propidium iodide staining using two-color flow cytometry with granulocyte-specific Gr1-FITC after 6-h whole blood culture with or without Fas agonist Jo2 (300 ng/ml) in the presence or absence of cycloheximide (CHX, 30 microg/ml). Membrane-associated receptors (Fas, TNFR-p55 and TNFR-p75) and cytoplasmic A1 expression of freshly isolated neutrophils were assessed by one-color flow cytometry and western blotting respectively. Compared with the group-WT/Sal, Jo2 induced apoptosis only in the presence of CHX (J+C). J+C-induced apoptosis was significantly lower in the group-p55(-/-)/Sal and p55( /-)/p75(-/-)/Sal but not in the group-p75(-/-)/Sal. J+C-induced apoptosis was inhibited similarly in all the LPS-injected WT and KO mice. Strong A1 expression was also induced similarly in all the LPS-injected WT and KO mice. Fas and TNFR p55 expression was normal and TNFR-p75 was significantly increased in all the LPS injected WT and KO mice although absence of the appropriate surface receptors was confirmed in the KO mice. We conclude that p55 normally plays a proapoptotic role, but p75 appears to play a minimal role in Fas-mediated neutrophil apoptosis. During endotoxin-induced systemic inflammation, both TNFR-p55 and TNFR p75 appear to be of minimal importance for modulation of Fas-mediated apoptosis and associated A1 protein expression despite normal Fas/TNFR-p55 and increased TNFR-p75 expression in neutrophils. PMID- 12630519 TI - Hemorrhage-induced vascular hyporeactivity to norepinephrine in select vasculatures of rats and the roles of nitric oxide and endothelin. AB - Hemorrhage-induced vascular hyporeactivity to norepinephrine (NE) and the possible effector roles of nitric oxide (NO) and endothelin (ET) were investigated in different vascular beds of rats. Under urethane anesthesia, rats (n = 7 per group) were hemorrhaged to a mean arterial pressure (MAP) of 50 mm Hg for 60 min. A group of rats was pretreated with either NG-nitro-L-arginine methyl ester (10 mg/kg), an NO synthase inhibitor, or PD142893 (0.1 mg/kg), an ET receptor antagonist 15 min before the end of the hypotensive period. Operated, euvolemic rats served as controls. The responses of MAP and the blood flow of the superior mesenteric (SMA), celiac (CA), left renal (LRA), and left femoral arteries (LFA) to NE (3 microg/kg, i.v.) were measured at baseline (prehemorrhage), at the end of the hypotensive period (0 h), and at 1, 2, and 4 h after the end of the hypotensive period. The pressor responses to NE on MAP at 0, 1, 2, and 4 h in the 60-min hemorrhage groups were reduced to 45.9%, 37.8%, 29.2%, 18.4% of baseline pressor response, respectively. At these same times, the fall in blood flow in response to NE in SMA, CA, LRA, and LFA was significantly blunted (P < 0.01). This loss of responsiveness in CA and LFA was more severe than in SMA and LRA (P < 0.05-P < 0.01). Pretreatment with L-NAME or PD142893 significantly improved the pressor response of MAP and the blood flow responses of the four arteries to NE (P < 0.01). Hypotension at 50 mm Hg for 60 min resulted in an apparent loss of vascular reactivity to NE, and the four vasculatures studies were not affected to the same extent. In addition, NO and ET appear to contribute to the loss of vascular reactivity in different vasculatures in hemorrhagic shock. PMID- 12630520 TI - Sympathetic blockade in a canine model of gram-negative bacterial peritonitis. AB - We investigated, in a well-established canine model of human sepsis, the effects of two different techniques of sympathetic blockade during bacterial peritonitis on pain relief, hemodynamics, and survival rate. Twenty-two purpose-bred beagles (12-28 months old, weighing 10-12 kg) were studied. Fourteen animals received an epidural infusion of bupivicaine and morphine, and the other eight received either a celiac plexus block (n = 4) or a sham block (n = 4). Eighteen of the 22 animals received an intraperitoneal challenge of Escherichia coli (1-10 x 10(9) CFU kg(-1) body weight). At comparable doses of intraperitoneal-implanted E. coli (2.5-5 x 10(9) CFU kg(-1) body weight), the addition of sympathetic blockade produced a synergistic decrease in survival times (P = 0.002) and mean left ventricular ejection fraction (P = 0.008), and increase in creatinine levels (P = 0.02). There was also a significant increase in tumor necrosis factor (TNF) levels (P = 0.004) and decrease in blood endotoxin clearance (P = 0.006) associated with sympathetic blockade during sepsis. The celiac plexus-blocked animals had no improvement in pain scores, and subjectively looked clinically worse than animals with sepsis without a celiac plexus block. In contrast, the epidural block was effective in blocking the pain and discomfort associated with low lethality doses of intraperitoneal bacteria reflected by no increase in pain scores compared with animals not receiving bacterial challenge. This study shows that during severe bacterial peritonitis, maintenance of sympathetic tone irrespective of pain relief provided is necessary for clearance of bacterial toxins, control of proinflammatory mediator release, hemodynamic stability, and survival. PMID- 12630521 TI - Hemodynamic effects of glibenclamide during endotoxemia: contrasting findings in vitro versus in vivo. AB - The final common pathway involved in the cardiovascular alterations of septic shock is incompletely defined. The opening of KATP channels is associated with vasorelaxation and alterations in cardiac contractility. This event may be triggered during septic shock by increased nitric oxide (NO) production, by a decreased intracellular content of ATP, or by a change in the transmembrane electrical potential. In the present study, we assessed the effects of glibenclamide, an agent that blocks the opening of KATP channels in vitro, on the contractile response of rat aortic rings to norepinephrine, and in vivo in anesthetized dogs, with or without exposure to Escherichia coli endotoxin. In vitro, glibenclamide decreased the contractile response to norepinephrine in the presence of endotoxin, provided that the endothelium was intact. In vivo, administration of 0.15 mg/kg increased systemic vascular resistance (SVR) in the absence of endotoxin only, and increased myocardial performance. A higher dose of 1 mg/kg increased SVR and decreased myocardial performance, both during endotoxic shock and in control conditions. Renal and mesenteric blood flows decreased, but the respective fractional flows were unchanged. Oxygen delivery decreased in both experimental conditions, but oxygen consumption decreased only in control conditions. The in vitro observations suggest that the opening of KATP channels is involved in the regulation of vascular tone during endotoxemia, via an endothelium-dependent mechanism. As different effects of glibenclamide were observed in vivo, the importance of the opening of KATP channels in endotoxic shock may be limited. PMID- 12630522 TI - Proinflammatory cytokines cause NO*-dependent and -independent changes in expression and localization of tight junction proteins in intestinal epithelial cells. AB - Intestinal epithelial barrier function is impaired after the exposure of enterocytes to proinflammatory cytokines. The mechanism(s) responsible for this phenomenon remain incompletely understood. We used cultured monolayers of Caco-2 enterocyte-like cells to characterize the effect of cytomix, a mixture of interferon-gamma, tumor necrosis factor-alpha, and interleukin-1beta, on the expression and localization of several tight junctions proteins. Cells were stimulated with cytomix in the presence or absence of 2-(4-carboxyphenyl)-4,4,5,5 tetramethylimidazoline-1 -oxyl-3-oxide (cPTIO), an NO* scavenger. Some cells were treated with (Z)-1-[2-(2-aminoethyl)-N-(2-ammonioethyl) amino]diazen-1-ium-1,2 diolate] (DETA-NONOate), an NO* donor. Tight junction protein expression was measured in cellular extracts by Western blotting and localized in cells using immunofluorescence. Steady-state mRNA levels were determined using semi quantitative reverse-transcription polymerase chain reaction. Incubation of cells with DETA-NONOate or cytomix decreased epithelial barrier function, decreased expression of ZO-1 mRNA, decreased expression of ZO-1, ZO-3, and occludin protein, and increased expression of claudin-1 protein. The effects of cytomix on barrier function and tight junction protein expression were modulated by cPTIO. Cytomix caused incorrect subcellular localization of ZO-1, occludin, and claudin 1, and this was modulated by co-incubation with cPTIO. DETA-NONOate caused similar protein mislocalization as observed with cytomix. The effectiveness of cPTIO in maintaining tight junction protein expression and correct subcellular localization was less apparent at early time points (12 h) compared with later points, suggesting an NO*-independent effect of cytokines on barrier function. Thus, cytomix appears to increase the permeability of Caco-2 monolayers through NO*-dependent and -independent mechanisms that are associated with changes in the expression and/or targeting of proteins involved in tight junction function. PMID- 12630523 TI - Analysis of tyrosine phosphorylation in resident peritoneal cells during diet restriction by laser scanning cytometry. AB - Tyrosine phosphorylation plays a critical role in signal transduction pathways in immune cells. Laser scanning cytometer (LSC), a newly developed microscope-based cytofluorometer, may overcome shortcomings of Western blotting and flow cytometry in the detection of intracellular signaling transduction. The aims of this study were to visualize and quantitate intracellular phosphotyrosine in the peritoneal cells harvested from diet-restricted mice by LSC. In addition, using LSC, we identified the main cell type with activated tyrosine phosphorylation in response to an inflammatory stimulus and we investigated the intracellular distribution of tyrosine phosphorylation within the peritoneal macrophages. Mice were assigned to the ad libitum and diet-restricted, i.e., 75% restricted food intake, groups. After 7 days of pair feeding, the peritoneal cells were harvested. Tyrosine phosphorylation in the harvested cells with either N-formyl-methionyl-leucyle phenylalanine (fMLP) or lipopolysaccharide (LPS) stimulation was examined using LSC. Tyrosine phosphorylation of peritoneal cells from the diet-restricted group was significantly higher than that from the ad libitum group, regardless of stimulation. Stimulation of peritoneal cells with either fMLP or LPS significantly increased tyrosine phosphorylation in the ad libitum group, but not in the diet-restricted group. The relocation feature of LSC revealed that the cells with distinct tyrosine phosphorylation were macrophages. Topographic analysis demonstrated that phosphotyrosine was localized mainly in the cytoplasm of these cells. In summary, LSC revealed that tyrosine phosphorylation is mainly in the cytoplasm of the peritoneal macrophages and is deranged by diet restriction. LSC is a powerful tool for the study of intracellular signaling transduction. PMID- 12630524 TI - Protective effect of 3-deazaadenosine in a rat model of lipopolysaccharide induced myocardial dysfunction. AB - Severe sepsis is accompanied by a profound depression of myocardial contractility. Leukocyte adhesion with subsequent local excess nitric oxide and reactive oxygen species production play major roles for this deleterious effect. We hypothesized that 3-deazaadenosine (c3Ado), an adenosine analogue with anti inflammatory properties, prevents endotoxin-induced myocardial dysfunction. Wistar rats (8 per group) were treated with Escherichia coli lipopoly-saccharide (LPS, 1 mg/kg, i.p., strain 0111:B4) +/- c3Ado (10 mg/kg, i.p.) 8 h before their hearts were harvested for isolated perfusion, histochemical analysis, or electrophoretic mobility shift assay. LPS induced a marked depression of left ventricular contractility. Immunohistochemistry revealed an upregulation of the adhesion molecules VCAM-1, ICAM-1, and P-selectin within the postcapillary venules. c3Ado inhibited VCAM-1 and ICAM-1 upregulation, but not P-selectin, and prevented cardiodepression. Electrophoretic mobility shift assay revealed inactivation of the transcription factor nuclear factor-kappaB and immunohistochemical staining for gp91phox, ED1, and CD11b demonstrated that c3Ado prevented local recruitment of monocytes and polymorph nuclear neutrophils to the myocardium. Accordingly, significantly fewer leukocytes producing nitric oxide or reactive oxygen species accumulated within the myocardium. Intravital microscopy of intestinal venules confirmed that LPS-induced adhesion of leukocytes was prevented by c3Ado. Additionally, c3Ado prevented LPS-induced elevation of serum tumor necrosis factor-alpha levels. Our results imply that c3Ado may prove to have clinical relevance for inflammatory disease processes. PMID- 12630525 TI - Antioxidant amplifies antibiotic protection in the cecal ligation and puncture model of microbial sepsis through interleukin-10 production. AB - Preadministration of antioxidants such as pyrrolidine dithiocarbamate (PDTC) and phenyl N-tert-butyl nitrone (PBN) protects animals from lethality in sepsis models. However, the requirement of preadministration greatly diminishes the clinical significance of these studies. Although the synthetic antioxidant PBN has been shown to effectively protect rodents from lethality in endotoxemia (lipopolysaccharide [LPS] model), preliminary screening indicates that pre- or postadministration of PBN does not protect in the rat cecal ligation and puncture (CLP) model. We show in this report that in a rat CLP model, the administration of PBN (150 mg/kg, 30 min after CLP) followed by the antibiotic imipenem (IMP; 10 mg/kg, 1 h after CLP) significantly increased survival compared with other single treatment groups. Previously, we have shown that PBN's protection in a rat LPS model is mediated by the overproduction of the anti-inflammatory cytokine interleukin (IL)-10. We show in this study that the increase in survival found in the PBN + IMP-treated group was abrogated by immunoneutralization with anti-IL-10 antibody, indicating that endogenous IL-10 is an effective protective factor. Plasma LPS levels were shown to be elevated after imipenem treatment, and the increased LPS level could have assisted to overproduce endogenous IL-10, as in the case of the PBN-treated LPS model. Statistical analysis indicated that the increase of IL-10 in PBN + IMP-treated group at early time period has significant association to the improvement of survival. PMID- 12630526 TI - Hypoxia and extraintestinal dissemination of Candida albicans yeast forms. AB - Candida albicans is a pleomorphic fungus with budding yeast and filamentous forms, and is a frequent cause of complicating infections in patients who are postsurgical, in shock, and have trauma. Many cases of systemic candidiasis are thought to orginate from the intestine, but it is unclear if the filament or the yeast is the more invasive form. Because C. albicans is relatively noninvasive and because mesenteric ischemia is thought to facilitate extraintestinal microbial dissemination, wild-type C. albicans CAF2 and mutant HLC54 (defective in filament formation) were orally inoculated into antibiotic-treated mice that were housed exclusively in room air, or were intermittently exposed to 10% oxygen for 1-h intervals. Both strains of C. albicans colonized the cecum in similar numbers (approximately 10(6.7)/g). C. albicans translocation to the draining mesenteric lymph nodes was not detected in mice inoculated with CAF2 (normoxic or hypoxic) or in normoxic mice inoculated with HLC54, but was detected in 33% (P < 0.01) of hypoxic mice inoculated with HLC54. Using Caco-2 and HT-29 enterocytes cultivated on plastic dishes and pretreated for 48 h in 10% oxygen, adherence of C. albicans HLC54 was decreased compared with wild-type CAF2, and hypoxia had no noticeable effect on adherence of either CAF2 or HLC54. Using enterocytes cultivated on permeable 8-microm filters, transepithelial migration of C. albicans CAF2 and HLC54 appeared similar. Thus, C. albicans HLC54 (defective in filament formation) was more invasive in hypoxic mice compared with wild-type CAF2, and host factors (e.g., mesenteric ischemia) rather than an innate ability to interact with enterocytes might play a more important role in extraintestinal dissemination of C. albicans yeast forms. PMID- 12630527 TI - Hepatic killing but not clearance of systemically circulating bacteria is dependent upon peripheral leukocytes via Mac-1 (CD11b/CD18). AB - The hepatic reticuloendothelial system (RES) is the primary mechanism for removing circulating bacteria from the systemic circulation. While Kupffer cells are important for this process, leukocytes appear to play a significant role as well. Hepatic leukocyte accumulation following ischemia/reperfusion or cytokine stimulation is well documented, but its contribution to phagocytic killing by the hepatic RES is not fully understood. We evaluated the role of leukocytes in general, and leukocyte-endothelial adhesion in particular, in hepatic RES function. This was done by inducing confirmed leukopenia with cyclophosphamide or by blocking leukocyte-endothelial adhesion molecules with specific blocking antibodies. Hepatic phagocytic clearance (HPC) and hepatic phagocytic killing (HKE) of systemically intravenously injected E. coli were assayed and quantitated by a validated dual isotope label technique. HPC among the various experimental groups and respective controls varied only slightly, with no statistically significant differences observed. Leukopenia or CD11b blockade each significantly decreased the HKE relative to the controls. Antibody blockade of certain other adhesion molecules had no significant effect on HKE (or HPC). The role of leukocytes in killing systemically circulating bacteria is an integral component of hepatic RES function. This capability of the leukocyte appears to be dependent, in part, on the adhesion molecule, Mac-1. PMID- 12630528 TI - Effect of trauma-hemorrhagic shock on red blood cell deformability and shape. AB - Previous work in our laboratory has demonstrated a decrease in red blood cell (RBC) deformability in sepsis. This has not been studied following hemorrhagic shock. We tested the hypotheses that hemorrhagic shock, associated with soft tissue trauma, leads to decreased RBC deformability and that this is related to alterations in the resting shape of the RBC. Elongation index (EI), a measure of RBC deformability, was determined over a range of shear stresses from 0.3 to 30 Pa in 26 male rats before and at various times after 90 min of hemorrhagic shock. RBC resting shape was determined by scanning electron microscopy. The data demonstrate that EI decreased significantly at the end of shock (before resuscitation), and remained below normal throughout the 6-h postshock period. Eight of the 26 animals decompensated during shock, requiring return of a portion of the shed blood to maintain a mean arterial pressure of 30-40 mmHg. Four of eight decompensated animals died before the end of the study period, compared with none of the compensated rats. The decompensated rats had significantly lower EI at 0.3 Pa by the end of the shock period (0.050 +/- 0.009) than the compensated shock group (0.058 +/- 0.006; P < 0.05). RBC shape alterations were first demonstrated at the end of the shock period and persisted throughout the 6 h postshock resuscitation period. These data indicate that trauma and hemorrhagic shock cause RBC shape alterations and a significant decrease in RBC deformability, which becomes manifested during the shock period and persists for at least 6 h postshock. Additionally, a direct relationship appears to exist between the magnitude of the physiologic insult and the degree of RBC damage. PMID- 12630529 TI - Endothelium-dependent vasodilation in the skin microcirculation of patients with septic shock. AB - The evidence for endothelial dysfunction in sepsis is mostly restricted to animal models. We investigated endothelial function in the skin microcirculation of eight patients hospitalized for septic shock in an intensive care unit (ICU). All patients required adrenergic support. Twelve hemodynamically stable ICU patients without sepsis who did not receive any vasoactive medication were used as controls. The two groups were of similar age and sex ratio. For additional reference, 16 healthy, nonsmoking subjects matched for age and sex to the first two groups were also studied. The evaluation of endothelial function was based on the comparison of skin blood flow responses to iontophoretically applied acetylcholine (Ach, an endothelium-dependent vasodilator) and sodium nitroprusside (SNP, an endothelium-independent vasodilator). Skin blood flow was measured on the volar face of the forearm using laser Doppler imaging. Before application of Ach or SNP, the mean baseline skin blood flow was below 100 perfusion units (PU) in all subjects and did not differ between groups. The maximal increase in blood flow elicited by both agents was significantly depressed in the patients with sepsis (Ach: 167 +/- 63 PU; SNP: 138 +/- 34 PU, mean +/- SD) compared with the ICU control patients (Ach: 291 +/- 135 PU, P < 0.05; SNP: 261 +/- 121 PU, P < 0.01) and the healthy, nonsmoking groups (Ach: 336 +/- 98 PU, P < 0.01; SNP: 304 +/- 81 PU, P < 0.01). The ratio of responses to Ach and SNP did not significantly differ between groups (septic: 1.22 +/- 0.40; ICU control 1.18 +/- 0.46, healthy, nonsmoking 1.12 +/- 0.24, P = 0.86). Thus, sepsis was not associated with a selective depression of the endothelium-dependent response. These results suggest that the capacity of the endothelium to produce signals for vasorelaxation remains intact in the skin microcirculation of patients with septic shock. PMID- 12630530 TI - Inhibition by terbutaline of nitric oxide and superoxide anion levels of endotoxin-induced organs injury in the anesthetized rat. AB - Despite the fact that septic shock is characterized by a decrease in systemic vascular resistance, the main cause of death is due to multiple organ failure. The organ dysfunction is usually attributed to cell death caused by overproduction of free radicals derived from inflammation. In the host infected by endotoxin (lipopolysaccharide, LPS), the expression and release of proinflammatory tumor necrosis factor-alpha (TNF-alpha) rapidly increases, and the formation of free radicals (e.g., superoxide anion [O2*-] and nitric oxide [NO*] in the present study) are inevitably overproduced. In this study, we present evidence that overall treatment of LPS rats with terbutaline, a beta2 adrenoceptor agonist, attenuates the delayed hypotension and ameliorates the tachycardia. Overproduction of TNF-alpha and NO* (produced by inducible NO synthase [iNOS] examined by Western blot analysis in the lung and the liver) is inhibited by treatment of LPS rats with terbutaline. In addition, treatment of endotoxemic rats with terbutaline also reduces the O2*- levels in the lung and the liver. Terbutaline also improves the liver (assessed by aspartate aminotransferase, alanine aminotransferase, total bilirubin, and albumin/globulin) and kidney (assessed by creatinine and uric acid) dysfunction induced by endotoxin. These findings suggest that the amelioration of circulatory failure and organs injury by terbutaline is associated with its suppression in TNF-alpha, O2*- and NO (via iNOS) production in animals with endotoxic shock. PMID- 12630531 TI - Enalaprilat improves systemic and mesenteric blood flow during resuscitation from hemorrhagic shock in dogs. AB - We investigated the systemic and mesenteric cardiovascular effects of administering enalaprilat during resuscitation from hemorrhage. Dogs were hemorrhaged (mean arterial pressure [MAP] 40-45 mmHg for 30 min, then 30-35 mmHg for 30 min) and were then resuscitated with intermittent lactated Ringer's solution (200 mL/kg/h during first 40 min, and 60 mL/kg/h during the following 130 min, MAP 75-80 mmHg). A constant-rate infusion of saline with or without enalaprilat (0.02 mg/kg/h) was initiated after 40 min of resuscitation. Blood flows declined with hemorrhage, increased with resuscitation, and then declined during the initial 40 min of resuscitation. Enalaprilat administration resulted in blood flow increases not seen in the controls (ending values for cardiac index: 2.8 +/- 0.4 L/min/m2 vs. 1.6 +/- 0.3 L/min/m2; celiac arterial flow 314 +/ 66 L/min/m2 vs. 139 +/- 13 mL/min/m2; and portal venous flow 596 +/- 172 L/min/m2 vs. 414 +/- 81 mL/min/m2 for enalaprilat versus controls, respectively). The greater flows with enalaprilat appeared to be due to prevention of the increases in afterload noted in the controls (ending arterial elastance values 3.73 +/- 0.97 mmHg/m2/mL vs. 7.74 +/- 1.80 mmHg/m2/mL for enalaprilat versus controls, respectively). We conclude that administration of a constant-rate infusion of enalaprilat during resuscitation can be used to improve systemic and mesenteric blood flow. PMID- 12630532 TI - 'No touch' techniques for porcelain ascending aorta: comparison between cardiopulmonary bypass with femoral artery cannulation and off-pump myocardial revascularization. AB - BACKGROUND: Detection of severe atherosclerotic ascending aorta during coronary artery bypass grafting requires alterations in the standard surgical technique to reduce the probability of stroke-related atheroembolization. Off-pump coronary artery bypass grafting (OPCAB) confers the benefits of avoiding aortic cannulation and clamping, and may therefore attenuate this risk. METHODS: OPCAB (n = 41) was compared to cardiopulmonary bypass (CPB) using femoral arterial cannulation and hypothermic fibrillatory arrest (n = 15), in patients with porcelain ascending aorta undergoing myocardial revascularization. In both groups, a 'no touch' technique was applied by avoiding aortic cannulation and clamping. Proximal anastomoses on the atherosclerotic aorta were avoided by arterial grafting, (in-situ or T-graft configurations) in all cases. RESULTS: Operative mortality was comparable (2.4% and 6.6% in the OPCAB and CPB groups respectively, p = NS). The rate of adverse neurological events, (two strokes and one transient ischemic attack), was higher in the CPB group (p = 0.0164). Based on brain CT, the nature of the recorded stroke suggested retrograde emboli. Three year survival (Kaplan-Meier) for the OPCAB and CPB groups was 86.7% and 81.3%, respectively (p = NS). Occurrence of late neurological adverse events during follow-up (8-51 months) was similar. CONCLUSIONS: In patients with porcelain ascending aorta undergoing myocardial revascularization, neurological outcome of OPCAB patients is better than CPB using femoral artery cannulation. PMID- 12630534 TI - Redo off-pump coronary artery bypass grafting for high risk hemodialysis patients. AB - BACKGROUND: The number of reoperative (redo) coronary artery bypass grafting (CABG) for patients with long-term hemodialysis has been increasing. Off-pump CABG (OPCAB) may decrease risks associated with redo CABG. METHODS: Two patients on chronic hemodialysis with calcification of the ascending aorta underwent redo double coronary OPCAB for the left anterior descending artery (LAD) and the posterior descending artery (PDA) via median sternotomy. The LAD was bypassed with the left internal thoracic artery (LITA). The PDA was exposed with minimum dissection and bypassed with a composite graft of the right internal thoracic artery (RITA) and the saphenous vein (SV). RESULTS: Both patients made a quick recovery with no complications and one had postoperative angiography that showed the patent grafts. Both patients were free from angina pectoris at follow-up of 6 months and 3 months, respectively. CONCLUSION: Redo OPCAB of the LAD and PDA can be performed with minimal dissection via median sternotomy using the LITA and a composite graft of the RITA and SV. PMID- 12630533 TI - Off-pump coronary artery bypass grafting in patients with end-stage renal disease on hemodialysis. AB - BACKGROUND: Coronary artery bypass grafting (CABG) for hemodialysis patients is high risk compared with other patient groups. The aim of this study was to analyze the potential benefits of off-pump CABG for hemodialysis patients. METHODS: From April 1994 through December 2000, 26 hemodialysis patients underwent CABG. The off-pump group consisted of 15 patients operated on without a pump and the on-pump group consisted of 11 patients operated on with a pump. RESULTS: There was no difference between the two groups with regard to mean age, mean number of diseased vessels and mean number of anastomoses per patient. No patient died in either group during hospitalization. The postoperative complication rate was low in both groups. The postoperative ventilation time was shorter in the off-pump group (8.5 vs 26.1 hours, p < 0.001, respectively [off pump group vs on-pump group]). The length of ICU stay was shorter in the off-pump group (1.7 vs 3.5 days, p = 0.01, respectively [off-pump group vs on-pump group]). The medial cost was lower in the off-pump group (26,200.80 dollars versus 44,024.10 dollars p = 0.0001 respectively [off-pump group vs on-pump group]). CONCLUSIONS: Off-pump CABG provided excellent less-invasive cardiac surgical results for dialysis patients. PMID- 12630535 TI - Thrombolytic therapy with recombinant streptokinase for prosthetic valve thrombosis. AB - BACKGROUND: Thrombosis is a serious complication of prosthetic heart valves, and management is often difficult. Thrombolytic therapy is a promising alternative to valve re-operation in the prosthetic valve thrombosis. METHODS: Fifteen consecutive patients with prosthetic heart valve thrombosis (10 mitral, 3 aortic, 2 tricuspid) were treated with intravenous recombinant streptokinase: 250,000 UI given over 30 minutes followed by an infusion an 100,000 UI per hour, always with clinical monitoring and echocardiographic examinations repeated at 24, 48, and 72 hours after starting thrombolytic therapy. Doppler echocardiography was the primary method use for diagnosis and was also used to follow the response to therapy RESULTS: Fibrinolytic treatment was successful in 14 (93.3%) patients. Total response was achieved in 13 (86.6%)patients and partial response in 1 (6.7%) patient; one patient died of ventricular fibrillation. No major hemorrhagic events were observed, peripheral embolism occurred in two cases, and one case of minor peripheral bleeding occurred in another. Some patients experienced fever and chills. CONCLUSIONS: The present study demonstrates the feasibility, safety and efficacy of thrombolytic therapy, which may be considered as first-line therapy for prosthetic heart valve thrombosis. PMID- 12630536 TI - Pedicled LIMA graft to the posterior coronary artery: which side to be anastomosed, dorsal or ventral? AB - It has not clearly been defined in the literature which side of the pedicled LIMA graft should be anastomosed to the posterior coronary arteries. Using the ventral side of pedicled LIMA graft causes an 180 degree torsion of the pedicle. Sometimes this torsion may cause reduction or cessation of LIMA flow as we have seen in two of our patients. In this paper we point out that using the dorsal side of the pedicled LIMA graft for anastomosis to the posterior coronary arteries is helpful. PMID- 12630537 TI - Aortic root endoscopy in pediatric cardiac operations for aortic valvuloplasty. AB - We use aortic root endoscopy for assessment of the aortic valve in pediatric patients. A flexible fiberscope inserted through the ascending aorta provides clear and precise visualization of the aortic valve. This technique of endoscopic assessment will help to judge the cusp prolapse and malcoaptation of the aortic valve in pediatric aortic surgery. PMID- 12630538 TI - Transpericardial extra-anatomic intrathoracic aortic bypass conduits in the management of coarctation of the aorta. AB - Two varied cases of coarctation of the aorta are described, in which transpericardial ascending to descending thoracic aortic conduits were routed around the right atrium, utilizing deep hypothermic circulatory arrest. The approach was through a median sternotomy in two patients. The first case is a 52 year-old female who underwent combined aortic valve replacement for aortic stenosis and repair of aortic coarctation distal to the left subclavian artery. The second case was a fifth time reoperation in a 12-year-old girl with recurrent coarctating. Both patients remain well and asymptomatic. PMID- 12630539 TI - Results of the probe technique for transatrial repair of tetralogy of Fallot. AB - OBJECTIVE: Total correction of classical tetralogy of Fallot (TOF) by transatrial approach has become a standard procedure in the goal to minimize structural damage to the pulmonary pump. The most critical point in transatrial repair of TOF is infundibular dissection. Right atrial approach provides better surgical exposure for parietal extension of the infundibular septum when compared to the right ventricular approach. However it is not always easy to determine the localization and amount of muscle bundles to be resected and this surgical maneuver requires experience. METHODS: Nineteen patients who had repair of isolated TOF using this technique from 1993 to 2001 were reviewed. The mean age of patients were 5 +/- 2 years. Transatrial-transpulmonary approach were performed for all patients. To make easier the infundibular muscle bundles resection and to determine the localization and amount of muscle bundle to be resected, we placed a Hegar dilator into the right ventricle through pulmonary arteriotomy. The muscle bundles between the dilator and the anterior leaflet annulus of the tricuspid valve were totally excised until the intraventricular part of the dilator and pulmonary annulus became completely visible. The area between the Hegar dilator and the margins of the ventricular septal defect (VSD) was left untouched. None of the patients had transannular patch. To improve exposure, tricuspid valve detachment was performed in 11 patients. All patients were followed-up in our clinic every 6 months using echocardiography. RESULTS: There were no early or late deaths, and no reoperation for residual VSD or residual right ventricle (RV) outflow obstruction. All patients were in NYHA Class I. RV on the echocardiography was spared late dilatation and had a good late functional status. Eighteen patients had no or mild pulmonary regurgitation. One patient who had undergone tricuspid anterior leaflet detachment showed mild tricuspid insufficiency. CONCLUSION: On the basis of hemodynamic outcomes, this procedure for elective repair of TOF in selected cases shows excellent early and mid-term results. PMID- 12630540 TI - Concomitant pulmonary embolectomy and excision of a left atrial myxoma. AB - We report a rare case of concomitant pulmonary embolectomy and excision of a left atrial myxoma. A 64-year-old obese woman developed a pulmonary embolism 13 days after an operation for a brain tumor. Echocardiography revealed a 3-cm left atrial mass attached to the atrial septum. Urgent surgery, including pulmonary embolectomy and excision of the left atrial mass, was performed. Histopathology confirmed that the mass was a left atrial myxoma. The postoperative course was uneventful. PMID- 12630541 TI - Paraplegia after routine cardiac surgery: a rare complication. AB - Two cases of spinal cord infarction with clinical paraplegia are discussed in patients undergoing elective and uncomplicated cardiac procedures with cardiopulmonary bypass. In both cases, the primary etiology was likely peripheral embolization. Other possible etiologies are reviewed, previously reported cases are discussed, and possible solutions to avoid this devastating problem are addressed. PMID- 12630542 TI - Laser myocardial revascularization modulates expression of angiogenic, neuronal, and inflammatory cytokines in a porcine model of chronic myocardial ischemia. AB - BACKGROUND: Controversy exists whether transmyocardial laser revascularization (TMR) is associated with angiogenesis or neuromodulation and whether these are time-dependent phenomena. Accordingly, we performed a time-course analysis of the expression of angiogenic and neuronal factors following experimental percutaneous TMR. METHODS AND RESULTS: Five weeks after placing ameroid constrictors on the circumflex coronary artery, 16 pigs underwent left ventricular mapping guided TMR using Ho:YAG laser (2 J x 1 pulse) at 30 sites directed at the ischemic zones and 11 animals were ischemic controls. Histology and immunostaining were obtained at 1 and 2 weeks (4 TMR and 3 controls at each time point) and at 4 weeks (8 TMR and 5 controls) for vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), nerve growth factor (betaNGF), substance P (SP), and monocyte chemoattractant protein-1 (MCP-1). Immunoreactivity was scored using a digital image analysis system. Factor VIII staining was used for blood vessel counting. Enhanced regional expression of VEGF, bFGF and MCP-1 in the TMR group was noted at 1 and 2 weeks with a threefold increase at 4 weeks following TMR compared to controls. BetaNGF expression in the TMR group was enhanced at 1 and 2 weeks with subsequent decline at 4 weeks to the controls level. SP expression was not significantly different between groups at all time points. There was a twofold increase in the number of blood vessels in the TMR group at 4 weeks, which was not apparent earlier. CONCLUSIONS: These immunohistological findings suggest that cytokines expression compatible with angiogenesis and neuromodulation occurs early after TMR. Up-regulation of angiogenic and inflammatory cytokines may be more sustained than neuromodulation. PMID- 12630543 TI - Emergency coronary artery bypass surgery in the era of glycoprotein IIb/IIIa receptor antagonist use. AB - The use of intravenous glycoprotein (GP) IIb/IIIa platelet receptor antagonists in the management of patients with acute coronary syndrome or those undergoing percutaneous coronary intervention (PCI) has become increasingly common in recent years. There are three GP IIb/IIIa receptor antagonists currently available for clinical use. Patients on GP IIb/IIIa receptor antagonists who require emergency surgical revascularization may be at increased risk for excessive peri- and postoperative bleeding. The duration of action of eptifibatide and tirofiban are short because they bind reversibly to the GP IIb/IIIa receptor and have a short half-life. Therefore, within a relatively short time after discontinuation of these agents, surgery can be performed with little or no increased risk of bleeding and without the need for additional hemostatic measures. Abciximab has a short plasma half-life but a long duration of action due to its high-affinity binding of GP IIb/IIIa receptors. Early retrospective studies demonstrated a higher incidence of major bleeding and requirement for blood transfusion, especially in those undergoing surgery within 12 hours of the discontinuation of abciximab. However, platelet transfusion has been shown to successfully reduce the incidence of these complications. The current evidence therefore indicates that, with appropriate measures, urgent surgical revascularization can be safely performed in patients who have received a GP IIb/IIIa receptor antagonist with little added risk. The benefits of these agents in the treatment of patients with an acute coronary syndrome or undergoing PCI are not obviated by the need for emergency bypass surgery. PMID- 12630544 TI - Homocysteine, endothelial dysfunction, and coronary artery disease: emerging strategy for secondary prevention. AB - Atherosclerosis is an important medical problem of the 21st century, but traditional risk factors could only account for 50% of the problem. Hyperhomocysteinemia is emerging as an independent atherosclerosis risk factor, associated with folate deficiency, renal failure, and relative deficiency of MTHFR (C677T polymorphism) or other enzymes depending on gender, age, and smoking status. Hyperhomocysteinemia has been reported to occur in 11-22% of western people, in 3-5% of normal asymptomatic Chinese subjects aged 18-70 years in Hong Kong, Macau, Sydney, and San Francisco, 23-36% of Chinese in Hong Kong with premature coronary artery disease, and 29% of a nonselective series of coronary subjects in Hong Kong. Evidence is accumulating that documents its associations with atherosclerosis disease in both case-control observations and prospective cohort studies, in vitro experiments, and in vivo experimental models in both animals and human subjects, as well as the successful improvement by homocysteine lowering of endothelial function as surrogate atherosclerosis endpoints in asymptomatic human and coronary patients (secondary prevention). A number of large scale homocysteine-lowering trials are currently underway for stroke and heart attacks prevention. Collectively these trials will include more than 65,000 patients at high-risk for cardiovascular and stroke events, and should provide a reliable evidence-base for prevention. PMID- 12630545 TI - Synergism between prostanoids and other vasoactive agents. AB - BACKGROUND: Prostaglandin E2 is usually considered to be a vasodilator, but some vascular beds respond with weak vasoconstriction mediated by prostanoid EP3 receptors. We have used the guinea pig isolated thoracic aorta to examine the nature of the synergism between the EP3 agonist sulprostone and other vasoactive agents. METHODS: Muscle tension was recorded from endothelium-denuded rings of aorta suspended in conventional organ baths. Indomethacin and the TP-receptor antagonist GR 32191 were usually present. RESULTS: Sulprostone (0.1-300 nM) showed two profiles: low-responder preparations (maximum response = 15-35% of 100 nM U-46619 response) were insensitive to the L-type Ca2+ blocker nifedipine, whereas high-responders (maximum = 35-70%) showed a nifedipine-sensitive component at higher sulprostone concentrations only. Charybdotoxin (CTX), a blocker of large-conductance Ca2+-activated K+ channels (BKCa), slightly enhanced threshold sulprostone responses and markedly enhanced larger responses; the enhancements were abolished by nifedipine. In contrast, threshold sulprostone responses were dramatically enhanced in the presence of established small contractions to phenylephrine (alpha1-adrenoceptor agonist), U-46619 (TP agonist), cyclopiazonic acid (sarcoplasmic Ca2+ pump inhibitor), and 4 aminopyridine (4-AP, Kv channel blocker). Nifedipine had no effect on enhancements of threshold sulprostone responses, and partially inhibited the enhancements of larger responses. CONCLUSIONS: BKCa channel activation appears to increase progressively as sulprostone-induced contraction increases. CTX removes this "BKCa brake," thereby providing an "L-type channel" Ca2+ flux to prime the EP3-receptor-driven Ca2+-sensitization mechanism (via Rhokinase activation, unpublished observations). In contrast, the other agents, including 4-AP, direct a non-L-type channel source of Ca2+ to the calmodulin-myosin light chain arm. PMID- 12630547 TI - Myocardial protection: entering the new millennium. AB - Dual objectives at operation are technical success and absence of iatrogenic injury due to inadequate myocardial protection. We enter a new millennium, and the spectrum of surgical procedures used to correct abnormal structure is expanding. We need longer intervals of aortic clamping to make the correct diagnosis, and to implement a more natural correction (i.e., mitral valve repair, Ross procedure, aortic recontruction with stentless valves, homografts). Simultaneously, our patients have increased vulnerability to injury, so that growth is needed to advance our methods of protection, in the same way as we learn new operative techniques. This manuscript deals both with evolution of current methods and recognition of newer methods of protection, so that the dual relationship between protection and procedures will not separate. PMID- 12630548 TI - Beta-adrenergic stimulation modulation of heart rate via synchronization of ryanodine receptor Ca2+ release. AB - A robust "fight or flight response", largely mediated via acute beta-adrenergic receptor (beta-AR) stimulation to the heart to increase its beating rate and contractile performance, is an essential component of the vertebrate survival instinct. While it has long been recognized that activation of beta-AR increases the spontaneous beating rate of sinoatrial nodal cells (SANC), specific links between stimulation of beta-ARs and the resultant increase in firing rate have not been evaluated. Our recent studies employed imaging of subcellular Ca2+ release coupled with recording of membrane potential or current in single, isolated cardiac SANC, to seek novel links between beta-AR stimulation and ryanodine receptor Ca2+ release and heart rate. An overview of these recent results, which provides novel insights into mechanisms of cardiac reserve that underlie the "fight or flight instinct, is presented here. PMID- 12630546 TI - Matrix is the site of indirect effects of propranolol on the ion channelopathies in cardiac remodeling by L-thyroxine. AB - Cardiac remodeling by chronic L-thyroxine medication causes exaggerated cardiac arrhythmias in relation to ion channelopathies that involve multichannels. The matrix of lipid membrane is likely the key site where channel lesions, possibly will develop and be benefitted by drug intervention. Cardiac remodeling in rats and guinea pigs was developed by L-thyroxine 0.5 mg/kg SC for 10 days. Propranolol was instituted on days 8-10. Whole cell holding was applied to measure ion currents. An increase in HR, dispersion of QTc, mitochondrial Na+/K+ ATPase, Ca2+/Mg2+ ATPase, and LPO production were found in the model. T3 and T4 levels in plasma were high. Propranolol was effective in regressing cardiac remodeling, together with lowering all the parameters and the enhanced I(Ca.L),I(KS), and I(KR) currents, but T3 and T4 remained basically unchanged. The changes in ion channels are likely the consequence of the cardiac remodeling that is formed by oxidative stress and increased energy consumption provoked by L thyroxine. The benefit of propranolol on the disordered ion channels is mediated by its ability to ameliorate lesions of the matrix. PMID- 12630550 TI - Modulation of L-type Ca2+ channels by beta3-adrenoceptor activation and the involvement of nitric oxide. AB - BACKGROUND: Both beta1- and beta2-adrenoceptors (AR) in cardiac tissues are responsible for the excitatory effect of catecholamines. Recent evidence demonstrated the presence of another subtype of beta-AR (beta3-AR) in cardiac ventricular preparation. Activation of beta3-AR elicited a depressant response on ventricular contraction. The underlying mechanism(s) for the negative inotropism is relatively unknown. METHODS: We investigated the effects of beta3-AR activation on basal voltage-dependent Ca2+ channel (I(CaL)) amplitude of the guinea pig enzyme-dissociated single ventricular myocytes using amphotericin B (200 microg/mL) perforated-patch whole-cell patch-clamp techniques (approximately 22 degrees C). RESULTS: Application of (-)-isoprenaline ((-)-ISO) (100 nM, a nonselective beta-AR agonist) increased the basal I(CaL) amplitude (approximately 210% of control) (n = 8). However, in the presence of nadolol (1 degreesM, a beta1-/beta2-AR antagonist), the stimulatory effect of (-)-ISO on I(CaL )was abolished and a slowly developed inhibition of the basal I(CaL) was recorded (approximately 80% of control) (n = 9). A smaller degree of inhibition was observed with BRL 37344 (100 nM, a selective beta3-AR agonist) (58% of control) (n = 7). The inhibitory effect of (-)-ISO (with nadolol) and BRL 37344 persisted after washout. Pretreating the ventricular myocytes with L-NAME (0.3 microM, a nitric oxide synthase inhibitor), but not D-NAME (0.3 microM), abolished the inhibitory effect of (-)-ISO and BRL 37344 (n = 7-9). CONCLUSIONS: The results suggest that beta3-ARs are present in ventricular myocytes. Activation of the beta3-AR resulted in an inhibition of the basal I(CaL) amplitude probably due to the formation of nitric oxide. PMID- 12630549 TI - Role of kappa opioid receptor in cardioprotection of preconditioning: implications in cardiac surgery. AB - Kappa-opioid receptor mediates both immediate and delayed cardioprotection of ischemic/metabolic inhibition preconditioning. Both PKC and mitochondria ATP sensitive K+ channel, mediate the protective action of kappa-opioid receptor stimulation. In delayed cardioprotection, PKC-epsilon may produce its effect via mitochondria ATP-sensitive K+ channel, and a stress inducible heat stress protein 70 may also be involved. Metabolic inhibition causes cell death which is accompanied by Ca2+ overload and mobilization of Ca2+ from sarcoplasmic reticulum to cytoplasm. The altered Ca2+ homeostasis is restored partially by kappa-opioid receptor or metabolic inhibition preconditioning. The finding indicates that opioids such as morphine may be used in the treatment of ischemic heart diseases and in open heart surgery. PMID- 12630551 TI - Procaine in cardioplegia: the effect on EDHF-mediated function in porcine coronary arteries. AB - OBJECTIVES: Hyperkalemia in cardioplegia impairs the endothelium-derived hyperpolarizing factor (EDHF)-mediated function. This study examined the effect of procaine in cardioplegia on the EDHF-mediated response in porcine coronary arteries. METHODS: An isometric force study was performed in a myograph. Two rings taken from the same artery (diameter 200-450 microm) were incubated with Krebs solution (group I) or 20 mM K+ (group II) with/without procaine (1 mM) at 37 degrees C for 1 hour. The EDHF-mediated relaxation was induced by bradykinin (BK, -10 approximately -6.5 log M) after U46619 (-8 log M, in group I) or K+ precontraction (in group II) in the presence of indomethacin (7 microM), NG-nitro L-arginine (300 microM), and hemoglobin (20 microM). The membrane potential of a single smooth muscle cell was measured by a microelectrode after superfusion with Krebs solution with/without procaine for 1 hour. RESULTS: The EDHF-mediated relaxation was increased by the treatment with procaine with the EC50 shifted leftward (97.3 +/- 0.6% vs. 83.0 +/- 5.1% at -7 log M and 99.4 +/- 0.6% vs. 96.7 +/- 1.6% at -6.5 log M, p < 0.05; EC50: -8.57 +/- 0.24 vs. -7.92 +/- 0.23 log M, p < 0.05). Procaine decreased the BK-induced hyperpolarization from -72.3 +/- 0.7 mV to -68.8 +/- 0.8 mV (-6.5 log M, p < 0.01). The EDHF-mediated relaxation in arteries exposed to 20 mM K+ was not altered by procaine (49.9 +/- 7.4% vs. 55.8 +/- 7.6%, p > 0.05). CONCLUSIONS: In the coronary arteries, procaine has a depolarizing effect but it enhances EDHF-mediated relaxation. Addition of procaine in cardioplegia did not change the EDHF-mediated endothelial function. PMID- 12630552 TI - Effect of recombinant basic fibroblast growth factor (bFGF) on fibroblast-like cells from human rotator cuff tendon. AB - Rotator cuff tendon cells (RCC) derived from surgical samples showed fibroblast like morphology. Histological staining demonstrated collagen secretion by RCC. Immunohistological findings revealed that RCC secreted type I and III collagen, but not type II collagen. In addition, the SDS-PAGE analysis suggested that RCC predominantly produced type I collagen. Basic fibroblast growth factor (bFGF) had a stimulatory effect on the proliferation of RCC dose-dependently up to 1 ng/ml. Administration of bFGF suppressed the secretion of collagens from RCC in a dose dependent manner. PMID- 12630553 TI - Autonomic nervous functions in fetal type Minamata disease patients: assessment of heart rate variability. AB - In order to assess the cardiovascular autonomic nervous functions in patients with fetal type Minamata disease (FMD), we investigated blood pressure (BP), and conducted time and frequency domain analysis of heart rate variability (HRV). Subjects were 9 patients in Meisuien recognized as FMD, and 13 healthy age matched control subjects. HRV and BP were assessed after subjects rested in a supine position for 10 minutes. Electrocardiographic (ECG) data were collected for 3 minutes during natural breathing. Time domain analysis (the average of R-R intervals [Mean RR], standard deviation of R-R intervals [SD RR], coefficient of variation [CV]), and frequency domain analysis by fast Fourier transformation (FFT) (power of low frequency [LF] and high frequency [HF] component, expressed in normalized units[nu]) were then conducted. In the time domain analysis, the mean RR of the FMD group was significantly lower than that of the control group. Neither SD RR nor CV showed significant differences between the two groups, but both tended to be lower in the FMD group. In the frequency domain analysis, the HF component of the FMD group was significantly lower than that of the control group. Pulse pressure (PP) was significantly lower in the FMD subjects. These findings suggest that parasympathetic nervous dysfunction might exist in FMD patients, who were exposed to high doses of methylmercury (MeHg) during the prenatal period. Decrease of PP might be due to degenerative changes of blood vessels driven by exposure to high doses of MeHg. PMID- 12630554 TI - Measurement of phosphatidylcholine hydroperoxide in mild ischemia-reperfusion injury in rat intestine. AB - In the pathogenesis of intestinal ischemia-reperfusion injury, the measurement of lipid peroxides needs to be established. Sprague-Dawley rat intestines were assessed after 30 minutes of occlusion of the superior mesenteric artery followed by reperfusion at 30, 60, 120, 180, 360 minutes. Grade of the mucosal injury, accumulation of the activated neutrophils and ICAM-1 expression were transiently increased after reperfusion. Two measuring methods of mucosal lipid peroxides using thiobarbituric acid reacting substance (TBARS) and phosphatidylcholine hydroperoxide (PCOOH) were compared. PCOOH level was significantly increased after reperfusion, while the mucosal TBARS level showed no significant change. In conclusion, lipid peroxidation could be detected with high specificity and sensitivity by measuring the mucosal phosphatidylcholine hydroperoxide level. PMID- 12630555 TI - Complications of IgA nephropathy in a non-insulin-dependent diabetes model, the Akita mouse. AB - The Akita mouse, which has a mutation (Cys96Tyr) in the insulin 2 gene (Ins2(Akita)), is a model for diabetes. The male Akita mouse has diabetes, while females develop mild diabetes. This study aimed to investigate renal complications in the Akita mouse model, which has been maintained in a heterozygous state Ins2(Akita) (+/-) with a C57BL/6 background (B6). Renal function (BUN, creatinine), serum IgA concentrations and histological changes in the kidneys were evaluated in diabetic and control mice in a B6 background. Five each of male and female mice per group (diabetes vs. control) were killed at 10, 20, 30 and 40 weeks of age. The influence of major histocompatibility antigens (MHC) on renal complications was tested using male diabetic mice in a C3H/He (C3H) background. When compared with controls, diabetic males, but not females, developed impaired renal function with elevation of serum IgA after 30 weeks of age. Diabetic glomerulosclerosis advanced with age in both sexes. Diffuse granular mesangial deposits of IgA were detected by immunofluorescence microscopy in diabetic males after 20 weeks. We tested whether the IgA-associated lesions were influenced by MHC using diabetic males in a C3H background. Similar degrees of diabetic glomerulosclerosis and glomerular IgA deposition were found in mice with C3H and B6 backgrounds. Akita mouse is a unique mouse model showing both mesangial sclerosis and IgA nephropathy. PMID- 12630556 TI - Variable E-cadherin expression in a MNU-induced colon tumor model in rats which exposed with 50 Hz frequency sinusoidal magnetic field. AB - Inactivation of the E-cadherin system by multiple mechanisms, including both genetic and epigenetic events, plays a significant role in multistage carcinogenesis. We have investigated the effects of sinusoidal electromagnetic fields (SMF) on E-cadherin expression in an MNU (N-methyl-N-nitrosurea)-induced colon tumor model. Male wistar albino rats were used for the study. The rats were classified into four groups: I (MNU), II (SMF+MNU), III (SMF) and IV (control). After administered at MNU in 1st and 2nd groups, 2nd and 3rd groups were exposed to a sinusoidal magnetic field (SMF, 50 Hz, 5 mT) for 6 hours/day for 8 months. The expression of E-cadherin were examined in four groups of rat colon tissues by immunohistochemistry on paraffin sections. For immunohistochemical analysis, the labeled streptavidin biotin method was performed using a Vectastain Universal Quick Kit with microwave accentuation. Fisher's exact test was used for statistical analysis between proportions. Immunohistochemical studies of E cadherin expression in this model demonstrated significant differences for cytoplasmic expression pattern. These results suggest that the electromagnetic fields result in significant alterations in cell adhesion mechanisms. This study has implications for understanding the role of fields in cell detachment in cancer metastasis. Further work is required to determine the relative effect of the magnetic fields on these phenomena. PMID- 12630557 TI - Successful treatment for rhabdomyosarcoma by total spondylectomy in a child. AB - A 7-year-old girl with a retroperitoneal rhabdomyosarcoma having invaded the L3 vertebra was treated by combination therapy consisting of chemotherapy, surgical resection and intraoperative radiation. Surgically, total spondylectomy was performed through a combined anterior and posterior procedures, and the spine was reconstructed with fibula bone grafts using a pedicle screw system made of titanium alloy. Transverse junction plates were not used at operation because of their bulkiness for the patient. Five days postoperatively, the rods dislodged from the screws and the grafted bones leaned. A Chance fracture-like transverse fracture of the L4 vertebra occurred during the revisional operation. Transverse plates and sublaminar wires were used to reduce the fracture and achieved stability. Six years postoperatively, the spine was completely fused and no local recurrence or metastasis is detected. PMID- 12630558 TI - Insulin resistance and type 2 diabetes--an adipocentric view. AB - As a result of selecting triglycerides as the major vehicle for storing superfluous energy, evolution came up with a specialized cell type, the adipocyte, equipped to handle triglycerides and its potentially toxic metabolites -fatty acids. For the first time in history large human populations are subjected a wealth of cheap, accessible and palatable calories. This has created a situation, on a large scale not previously encountered, in which the capacity to store triglycerides in adipocytes is an important determinant of human health. Too few adipocytes (e.g. lipodystrophia) or a situation in which all adipocytes are filled, to their maximum capacity (e.g. severe obesity), will create very similar and unfavorable metabolic situations in which ectopic triglyceride stores will appear in tissues like liver and muscle. This review sets out to discuss the adipocyte and its role in metabolism as well as the consequences of a metabolic situation, in which the adipocyte has lost its fat storing monopoly. PMID- 12630560 TI - Chemokines in cardiovascular remodeling: clinical and therapeutic implications. AB - Chemokines are newly discovered molecules that mediate the migration of leukocytes into inflammed tissues and control the inflammatory reactions in various immune-mediated diseases. Both in animal models and in human specimens, chemokine expression is associated with atherosclerotic lesion development and vascular remodeling and restenosis after angioplasty. Furthermore, recent studies have demonstrated that chemokines play an important role in the pathophysiology of acute coronary syndromes, post-infarction left ventricular remodeling and chronic heart failure. The capacity to control activation and movement of inflammatory cells suggests that chemokines and their receptors might provide novel targets for therapeutic intervention in a number of conditions characterized by chronic inflammation, including cardiovascular diseases. The present review summarizes current knowledge regarding the potential pathogenic role of chemokines in major cardiovascular disorders, as well as the modulation of the chemokine network as a novel, interesting therapeutic modality in this field. PMID- 12630559 TI - Atopic dermatitis: molecular mechanisms, clinical aspects and new therapeutical approaches. AB - Atopic dermatitis (AD) is a genetically determinated, chronic inflammatory skin disorder associated with cutaneous erythema and severe pruritus, affecting 10-15% of children with increasing incidence and socio-economical relevance. Frequently, AD is associated with development of allergic rhinitis and/or asthma later in childhood. In most of patients AD is associated with a sensitization to food and/or environmental allergens and increased serum-IgE, while only a fewer percentage missed links to the classical atopic diathesis. Currently investigated pathogenetic aspects of AD include imbalanced Th1/Th2 responses, altered prostaglandin metabolism, intrinsic defects in the keratinocyte function, delayed eosinophil apoptosis, and IgE-mediated facilitated antigen presentation by epidermal dendritic cells. An inflammatory response of the two-phase-type and the effects of staphylococcal superantigens (SAgs) are also reported. At present a standardized cure of AD and a consensus on therapeutical approach of the severe form of the disease have not been established. Current management of AD is directed to the reduction of cutaneous inflammation and infection, mainly by S. aureus, and to the elimination of exacerbating factors (irritants, allergens, emotional stresses). Since patient with AD show abnormalities in immunoregulation, therapy directed to adjustment of their immune function could represent an alternative approach, particularly in the severe form of the disease. In this review, we analyse the clinical and genetic aspects of AD, the related molecular mechanisms, and the immunobiology of the disease, focusing our attention on current treatments and future perspectives on this topic. PMID- 12630561 TI - Potential mechanisms of cancer chemoprevention by anthocyanins. AB - Anthocyanins are the chemical components that give the intense color to many fruits and vegetables, such as blueberries, red cabbages and purple sweet potatoes. Epidemiological investigations have indicated that the moderate consumption of anthocyanin products such as red wine or bilberry extract is associated with a lower risk of cardiovascular disease and improvement of visual functions. Recently, there is increasing interesting in the pharmaceutical function of anthocyanins. This review summarizes current knowledge on the various molecular evidences of cancer chemoprevention by anthocyanins. These mechanisms can be subdivided into the following aspects: 1) the antioxidation; 2) the molecular mechanisms involved in anticarcinogenesis; 3) the molecular mechanisms involved in the apoptosis induction of tumor cells. Finally, the bioavailability and structure-activity relationship of anthocyanins are also summarized. PMID- 12630562 TI - Inflammatory mediators and the failing heart: a translational approach. AB - Recent studies have identified the importance of proinflammatory mediators in regulating cardiac structure in health and disease. Recent studies suggest that cytokines that are expressed within the myocardium in response to a environmental injury, namely tumor necrosis factor-alpha (TNF), interleukin-1 (IL-1) and the interleukin-6 (IL-6) family of cytokines play an important role in initiating and integrating homeostatic responses within the heart. However, these "stress activated" cytokines all have the potential to produce cardiac decompensation when expressed at sufficiently high concentrations. Indeed, there is now a growing appreciation that these molecules may play an important role in mediating disease progression in the failing heart. The growing appreciation of the pathophysiological consequences of sustained expression of proinflammatory mediators in pre-clinical and clinical heart failure models culminated in a series of multicenter clinical trials that utilized "targeted" approaches to neutralize tumor necrosis factor (TNF) in patients with moderate to advanced heart failure. However, these targeted approaches have resulted in worsening heart failure, thereby raising a number of important questions about what role, if any, proinflammatory cytokines play in the pathogenesis of heart failure. This review will summarize the tremendous growth of knowledge that has taken place in this field, with a focus on what we have learned from the negative clinical trials, as well as the potential direction of future research in this area. PMID- 12630563 TI - Pathologic mechanisms of influenza encephalitis with an abnormal expression of inflammatory cytokines and accumulation of mini-plasmin. AB - The pathogenesis of influenza encephalopathy or encephalitis is poorly understood. This review summarizes our recent studies of the roles played by inflammatory cytokines, inducible nitric oxide synthase (iNOS), adhesion molecules and mini-plasmin in influenza encephalitis. After the intranasal infection of newborn mice with the non-neurotropic strain of influenza A virus (IAV) Aichi/2/68/H3N2, encephalitis and severe brain edema were observed within 3 5 days. IAV-RNA and abnormalities in the blood-brain barrier permeability were detected in association with an increase in the mRNA expressions of endothelin-1, iNOS, and tumor necrosis factor-alpha. Furthermore, the accumulation in the brain capillaries of mini-plasmin, which proteolytically induces the viral envelope fusion activity and allows the virus to enter the cells, changes the brain from non-susceptible to susceptible to non-neurotropic IAV multiplication. The accumulation of mini-plasmin was markedly greater in newborn mice with an impaired mitochondrial fatty acid metabolism. These inflammatory mediators and the accumulation of mini-plasmin in the brain may play an important role in the onset and progression of LAV encephalitis. PMID- 12630564 TI - Toll-like receptor signaling in anti-cancer immunity. AB - It is important to augment the anti-cancer host response in cancer treatment. Recent studies suggested that the signaling via Toll-like receptors (TLRs) which are newly identified receptor molecules recognizing many pathogens, are involved in the induction of anti-cancer immunity. Seya et al. demonstrated that maturation of dendritic cells (DCs) and cytokine induction by the cell wall skeleton of Mycobacterium bovis bacillus Calmette-Guerin (BCG-CWS) are induced via both TLR2 and TLR4. Akira et al. discovered a new molecule of TLR family, TLR9, recognizing unmethylated bacterial CpG-DNA, whose clinical use is expected for cancer therapy as a potent inducer of a helper T cell 1 (Th1)-type T-cell response. TLR9-deficient mice did not show any responses to CpG-DNA, including Th 1 cytokine production and maturation of DCs. We have obtained two molecules, a lipoteichoic acid-related molecule isolated from streptococcal agent OK-432, and a plant-derived 55-kDa protein that can induce Th1 response and elicit a strong anti-cancer effect in vivo and in vitro. Our basic experiments demonstrate that TLR4 signaling is intimately involved in anti-cancer immunity induced by these immunopotentiators. Our clinical examination in oral cancer patients also suggests the requirement of both TLR4 and MD-2 in the OK-432-induced anti-cancer host response. Establishment and clinical use of the methodology for human cancer therapy by utilizing TLR signaling is greatly expected. PMID- 12630565 TI - Genetic polymorphisms of serotonin and dopamine transporters in mental disorders. AB - Transporter-assisted uptake of serotonin (5-HT) and dopamine (DA) has been accounted for activities in human behavior or mental status, because they are the sites of action of widely used antidepressant and psychoactive drugs. Both the human serotonin transporter (5-HTT) and human dopamine transporter (DAT1) genes are good candidates for etiological involvement in various psychiatric conditions. The serotonin transporter gene has two types of functional polymorphisms. One is serotonin transporter linked polymorphic region (5-HTTLPR) consisting of length variation of the repetitive sequence containing 20-23-bp long repeat elements in the 5'-upstream region of the gene. Another polymorphism is that serotonin transporter variable number of tandem repeats (5-HTTVNTR) in its second intron. Both polymorphisms affect the transcription ratio of 5-HTT gene and may modify neuronal transmission by changing its protein expression. On the other hand, DAT1 gene has a variable number of tandem repeats type polymorphism (DAT1VNTR) in the 3'-untranslated region of the mRNA, which was also reported to change its gene expression. So polymorphic variations of transporters would change the behavioral and neuropathological tendency. Here, the feature of those two transporters and their relations to psychiatric disorders are described. PMID- 12630566 TI - The role of caspase cascade on the development of primary Sjogren's syndrome. AB - Primary Sjogren syndrome (SS) is an autoimmune disease characterized by diffuse lymphoid cell infiltrates in the salivary and lacrimal glands, resulting in symptoms of dry eye and dry mouth due to insufficient secretion. Previously, we have identified the 120 kDa alpha-fodrin as an important autoantigen on the development of SS in both animal model and SS patients, but the mechanism of a fodrin cleavage leading to tissue destruction in SS remains unclear. In murine primary SS model, tissue-infiltrating CD4+ T cells purified from the salivary glands bear a large proportion of Fas ligand (FasL), and the salivary gland duct cells constitutively possess Fas. Infiltrating CD4+ T cells identified significant 51Cr release against mouse salivary gland (MSG) cells. In vitro studies demonstrated that apoptotic MSG cells result in a specific alpha-fodrin cleavage into 120 kDa, and preincubation with caspase-inhibitor peptides blocked alpha-fodrin cleavage. The treatment with caspase-inhibitors in vivo prevented the development of autoimmune lesions in the salivary and lacrimal glands. Thus, an increased activity in caspase cascade may be involved in the progression of alpha-fodrin proteolysis and tissue destruction on the development of SS. PMID- 12630568 TI - Long-term follow-up of gastric metaplasia after eradication of Helicobacter pylori. AB - BACKGROUNDS AND AIMS: There is no commonly accepted view concerning changes in gastric metaplasia after the eradication of Helicobacter pylori. The aim of this study was to evaluate the long-term course of gastric metaplasia after the eradication of this bacterium. METHODS: The subjects were 59 patients with duodenal ulcer who were positive for Helicobacter pylori. Forty patients were classified as the eradication group. Gastric metaplasia was endoscopically and histologically evaluated before and after eradication of this bacterium. The follow-up period was 2-7.1 years. In the other 19 patients in the non-eradication group, gastric metaplasia was evaluated before and after treatment of the ulcer. Gastric metaplasia was evaluated in terms of its extent and type in all patients. RESULTS: Gastric metaplasia showed the incomplete type before eradication but changed to the complete type after eradication, which persisted for a long period. The extent of gastric metaplasia increased after eradication. In the non eradication group, gastric metaplasia infrequently changed to the complete type during the scarring period of ulcer. CONCLUSION: Gastric metaplasia changed to the complete type after the eradication of Helicobacter pylori, which persisted for a long period. PMID- 12630567 TI - Isolation and characterization of a novel gene sfig in rat skeletal muscle up regulated by spaceflight (STS-90). AB - We obtained the skeletal muscle of rats exposed to weightless conditions during a 16-day-spaceflight (STS-90). By using a differential display technique, we identified 6 up-regulated and 3 down-regulated genes in the gastrocnemius muscle of the spaceflight rats, as compared to the ground control. The up-regulated genes included those coding Casitas B-lineage lymphoma-b, insulin growth factor binding protein-1, titin and mitochondrial gene 16 S rRNA and two novel genes (function unknown). The down-regulated genes included those encoding RNA polymerase II elongation factor-like protein, NADH dehydrogenase and one novel gene (function unknown). In the present study, we isolated and characterized one of two novel muscle genes that were remarkably up-regulated by spaceflight. The deduced amino acid sequence of the spaceflight-induced gene (sfig) comprises 86 amino acid residues and is well conserved from Drosophila to Homo sapiens. A putative leucine-zipper structure located at the N-terminal region of sfig suggests that this gene may encode a transcription factor. The up-regulated expression of this gene, confirmed by Northern blot analysis, was observed not only in the muscles of spaceflight rats but also in the muscles of tail-suspended rats, especially in the early stage of tail-suspension when gastrocnemius muscle atrophy initiated. The gene was predominantly expressed in the kidney, liver, small intestine and heart. When rat myoblastic L6 cells were grown to 100% confluence in the cell culture system, the expression of sfig was detected regardless of the cell differentiation state. These results suggest that spaceflight has many genetic effects on rat skeletal muscle. PMID- 12630569 TI - Study of life satisfaction and quality of life of patients receiving home oxygen therapy. AB - An investigation was conducted by mail using a questionnaire regarding the life satisfaction and quality of life (QOL) of patients receiving home oxygen therapy (HOT) to evaluate their support. QOL was evaluated according to 4 scales: (1) activities, (2) state of health and quality of living, (3) physical symptoms, and (4) economic state. The answers of 90 patients (recovery rate: 60%) who responded to the investigation were analyzed, and the following points were clarified. 1. Most of the subjects visited the hospital regularly, and about half the subjects (50.6%) had been treated by hospitalization during the 3 years prior to the investigation. 2. A large majority of the subjects (77.4%) answered they were satisfied with life. 3. Life satisfaction was closely related to the patients' roles and hobbies, and their activities in their communities and families. 4. The quality of living and the state of health were closely related to mental activity. 5. The economic state was closely related to all items of life satisfaction, quality of living, and state of health. From these results, expansion of the range of activities of patients receiving HOT and providing an economic basis for their living as well as preventing exacerbation of the disease are considered to be important for improving their life satisfaction. PMID- 12630570 TI - Clinical value of the determination of serum guanase activity in patients with chronic hepatitis type C. AB - The study examines the clinical significance of guanase (GU) measurement in patients with hepatitis C. 688 patients in whom either ALT was abnormal, or in whom HBsAg or HCVAb was detected in the serum, were enrolled into this study. The percentage of cases in which normal ALT while elevated GU was compared among the different disease groups. Then, the percentage of cases with normal ALT but elevated GU was compared between HBV and HCV groups. For the entire population, a significant correlation was observed between ALT and GU (r=0.872). The overall percentage of cases with normal ALT but elevated GU activity was 11.4%. In HCV group, 449 cases had normal ALT. Of these cases, 20.3% had elevated GU, while ALT was normal. Before 1989, no test to check donated blood for HCV antibody was available. However, screening of donated blood for high GU was associated with a reduced incidence of post-transfusion hepatitis. This is probably because following the screening, blood donated by patients with hepatitis C who had normal ALT but elevated GU was rejected. After the introduction of HCV antibody measurement, GU measurement is still useful to reveal the pathophysiological condition in-patients with chronic hepatitis type C. PMID- 12630571 TI - Inhaled steroid therapy and hospitalization for bronchial asthma: trend in Tokushima University Hospital. AB - With the recognition that airway inflammation is present even in patients with mild bronchial asthma, therapy with inhaled corticosteroids is now indicated in various stages of patients. In the present article, we retrospectively examined the prescriptions for inhaled corticosteroids and other drugs for the treatment of outpatients with bronchial asthma at Tokushima University Hospital. We also analyzed asthma control in these patients, in terms of the incidence of emergency consultations and hospitalizations due to asthma exacerbations. To analyze the recent trend, the patients observed from 1998 to 2000 (recent years) were included, and for control purpose, those in 1990 and 1991 (earlier years) were also included. The percentage of patients treated with inhaled corticosteroids remarkably increased in recent years (mean; 81.3%) compared to earlier years (mean; 23.5%). In contrast, the usage of oral corticosteroids, oral xanthine derivatives, beta-adrenergic receptor agonists and anti-allergic agents tended to decrease in the 10 years period. After the introduction in 1995, considerable patients up to 25% have been treated with anti-leukotrienes. Emergency consultations decreased in recent years (mean; 0.18/patient/year) compared to earlier years (mean; 0.79/patient/year). Emergency hospitalizations also decreased in recent years (mean; 0.043/patient/year) compared to earlier years (mean; 0.23/patient/ year). In the present study, spread of inhaled corticosteroid therapy and decline in incidence of emergency consultation and hospitalization were simultaneously observed at Tokushima University Hospital, and the former has, at least in part, a contribution to the latter. PMID- 12630572 TI - Cloning, expression analysis, and tissue distribution of esp-1/testisin, a membrane-type serine protease from the rat. AB - Esp-1/testisin, a serine protease abundantly expressed in human and mouse testis, is presumed to play an important role in the process of spermatogenesis and fertilization. In this study, we cloned an esp-1/testisin cDNA from rats, and analyzed its expression and tissue distribution. The isolated cDNA consisted of 1099 nucleotides with a single open reading frame encoding 328 amino acids and an expected molecular mass of 36.6 kDa. The deduced amino acid sequence of rat Esp 1/Testisin had 89% and 62% identity with its murine and human counterparts, respectively, and appeared to be a trypsin-type serine protease with a hydrophobic region at the C-terminus. By quantitative real-time polymerase chain reaction analysis, rat esp-1/testisin mRNA was predominantly expressed in testis, as in human and mouse. However, its immunohistochemical distribution was predominantly in the elongated spermatids at steps 12 to 19, and not in the primary spermatocytes and round spermatids. This different distribution profile suggests that Esp-1/Testisin plays a role in species-specific proteolytic events during spermatogenesis and fertilization. PMID- 12630574 TI - Effect of human airway trypsin-like protease on intracellular free Ca2+ concentration in human bronchial epithelial cells. AB - It has been shown that human airway trypsin-like protease (HAT) is localized in human bronchial epithelial cells (HBEC), and trypsin activates protease-activated receptor-2 (PAR-2). Activation of PAR-2 activates G-protein followed by an increase of intracellular free Ca2+, [Ca2+]in. This study was undertaken to clarify whether HAT can activate PAR-2 in HBEC or not. RT-PCR showed that HAT mRNA is expressed in HBEC, and PAR-2 mRNA is the most strongly expressed of the known PARs in HBEC. Both PAR-2 agonist peptide (PAR-2 AP) and HAT increased [Ca2+]in in HBEC in a biphasic fashion; a prompt, sharp increase (peak I) and a sustained low plateau (peak II). PAR-2 AP over 100-200 microM and HAT over 200 300 mU/ml (0.08-0.12 microM) induced both peak I and II, and PAR-2 AP below 100 microM and HAT below 200 mU/ml induced only peak II. Both PAR-2 AP-induced and HAT-induced peak I were induced by Ca2+ mobilization from intracellular stores, because they appeared even in Ca2+-free medium. Both PAR-2 AP-induced and HAT induced peak II were induced by an influx of extracellular Ca2+, because they were abolished in Ca2+-free medium. The Ca2+ response to HAT was desensitized by exposure of HBEC to PAR-2 AP. These results indicate that HBEC have a functional PAR-2, and HAT regulates cellular functions of HBEC via activation of PAR-2. PMID- 12630573 TI - Effect of endothelin-1 (1-31) on the renal resistance vessels. AB - Human chymase produces not only angiotensin II but also endothelin(ET)-1(1-31). We previously reported that ET-1(1-31) had several biological activities in vascular smooth muscle cells. In this study, we investigated the vasoconstrictor effect of ET-1(1-31) on the renal resistance vessels using in vitro microperfused rabbit afferent and efferent arterioles. ET-1(1-31) decreased the lumen diameter of the afferent and efferent arterioles dose-dependently. ET-1(1-31)-induced afferent arteriolar vasoconstriction was not affected by phosphoramidon, an ET converting enzyme inhibitor. ET-1(1-31)-induced renal arteriolar vasoconstriction was inhibited by BQ123, an ETA receptor inhibitor, but not by BQ788, an ETB receptor inhibitor. These results suggest that ET-1(1-31)-induced renal arteriolar vasoconstriction may be mediated by ETA-like receptors. PMID- 12630575 TI - Repair of incisional hernia with prolene hernia system. AB - A 70-year-old woman was admitted to our hospital with a complaint of bulging in the right lower portion of the abdomen. The bulging was in accordance with an old operative scar for appendicitis. The findings of computed tomography (CT) showed defects in the abdominal muscles and the protrusion of the intestine into the subcutaneous fat. The patient was diagnosed with incisional hernia after appendectomy and underwent a repair of the incisional hernia, using the prolene hernia system double-layer mesh. The patient's post-operative course was excellent. Recently, the prolene hernia system, double-layer mesh was reported to be effective for groin hernias due to its advantageous protection the recurrence through reinforcement of the patient's myopectrial orifice. It is suggested that this new device is also useful for small incisional hernias. PMID- 12630576 TI - The application of immunohistochemical findings in the diagnosis in methamphetamine-related death-two forensic autopsy cases-. AB - Forensic autopsy cases detecting methamphetamine (MA) are usually diagnosed according to its toxicological concentration. It has been reported that the lethal blood concentration of MA is 4.48 microg/ml (3.0 micromol/dl). We autopsied two MA-detected cadavers, and immunohistochemical staining was performed on the skeletal muscle with an anti-myoglobin antibody, and on the kidney with an anti-the 70 kDa heat shock protein (HSP70) antibody. One case showed a high rectal temperature (40 degrees C). The toxicological examination revealed 0.75 microg/ml of MA in the blood, and 16.8 microg/ml in the urine. Myoglobin was negative and HSP70 was positive in the kidney immunohistochemically. From the toxicological and immunohistochemical findings, it was considered that the subject died of hyperthermia and acidosis caused by muscular hyperactivity. In another case, the autopsy revealed highly congested lungs, with dark-red bloody fluid and foam in the trachea and bronchus. MA (17.0 microg/ml) was detected in the blood. HSP70 was negative and myoglobin was positive immunohistochemically. It was thought that the subject died of acute MA intoxication based on the high MA concentration, although rhabdomyolysis was suspected. It is suggested that myoglobin and HSP70 immunostaining are useful to diagnose MA poisoning. PMID- 12630577 TI - Discrepant results regarding long-term survival of patients with anorexia nervosa? PMID- 12630578 TI - Orthopedic surgery: a new frontier. PMID- 12630579 TI - Long-term survival of patients with anorexia nervosa: a population-based study in Rochester, Minn. AB - OBJECTIVE: To estimate long-term survival of unselected patients with anorexia nervosa from Rochester, Minn. PATIENTS AND METHODS: In this population-based retrospective cohort study, all 208 Rochester residents who presented with anorexia nervosa (193 women and 15 men) for the first time from 1935 through 1989 were monitored for up to 63 years. Subsequent survival was compared with that expected for Minnesota white residents of similar age and sex, and standardized mortality ratios were determined on the basis of age- and sex-specific death rates for the US population in 1987. RESULTS: Survival was not worse than expected in this cohort (P = .16). The estimated survival 30 years after the initial diagnosis of anorexia nervosa was 93% (95% confidence interval, 88%-97%) compared with an expected 94%. During 5646 person-years of follow-up (median, 22 years per patient), 17 deaths occurred (14 women and 3 men) compared with an expected 23.7 deaths (standardized mortality ratio, 0.71; 95% confidence interval, 0.42-1.09). One woman died of complications of anorexia nervosa, 2 women committed suicide, and 6 patients (5 women and 1 man) died of complications of alcoholism. Other causes of death were not increased. CONCLUSIONS: Long-term survival of Rochester patients with anorexia nervosa did not differ from that expected. This finding suggests that overall mortality was not increased among the spectrum of cases representative of the community. PMID- 12630580 TI - Primary and revision total hip arthroplasty for patients 90 years of age and older. AB - OBJECTIVE: To assess the reliability, durability, and safety of primary and revision total hip arthroplasty (THA) in patients 90 years of age and older. PATIENTS AND METHODS: Between 1970 and 1997, 66 THAs, 48 primary and 18 revision, were done for 65 patients aged 90 to 104 years (mean, 92 years). The Harris hip score was used to assess hip pain and function. RESULTS: Patients were evaluated for pain, function, and satisfaction at a mean of 3.2 years after the THA. The Harris hip scores improved significantly (P = .004) for both primary and revision groups: from preoperative mean scores of 39 and 45 points to follow-up scores of 80 and 75 points, respectively. Two patients died in the early postoperative period of causes related to the operation. Thirty-seven other patients were dead at last follow-up and had lived a mean +/- SD of 6 +/- 2.2 years after the primary THA and 4.2 +/- 2.8 years after the revision THA. No hips required subsequent revision. CONCLUSION: Total hip replacement was reliable, durable, and safe in this study group. Primary care physicians and surgeons should be aware that both primary and revision total hip replacement can be done safely and effectively in patients aged 90 years and older and can result in years of relief of pain and functional improvement. PMID- 12630581 TI - Validation and modification of streptococcal pharyngitis clinical prediction rules. AB - OBJECTIVE: To validate a simplified version of the Walsh clinical prediction rules (CPRs) for the presence of streptococcal pharyngitis in an inner-city, ethnically diverse population. PATIENTS AND METHODS: This prospective study conducted in New York City, NY, from January 1,1997, to May 31,1997, consisted of 171 consecutive adult walk-in patients who presented with symptoms of upper respiratory tract infection and/or sore throat. The patients were assessed by using 5 clinical factors: cough, exposure to known streptococcal contact, temperature, tonsillar-pharyngeal exudates, and cervical lymphadenopathy. Throat cultures for group A beta-hemolytic streptococcus were obtained from all patients. Clinicians assessing the patients were unaware of throat culture results, and those processing the throat cultures were blinded to the clinical predictors. RESULTS: The prevalence of streptococcal pharyngitis was 24% (95% confidence interval, 18%-30%). The simplified version of the Walsh CPR for streptococcal pharyngitis predicted accurately the probability of a positive culture in our diverse population (area under the receiver operating characteristic curve, 0.71). The simplified CPR also showed clinically useful likelihood ratios and posterior probabilities. CONCLUSION: A simplified version of the Walsh CPR is accurate for diagnosing streptococcal pharyngitis in an inner city population. This finding should provide clinicians more confidence in applying the CPR in similar clinical settings. PMID- 12630582 TI - Antiphospholipid antibodies and the risk of thromboembolic events in valvular heart disease. AB - OBJECTIVE: To investigate the role of antiphospholipid antibodies (aPLs) in subsequent thromboembolic events and mortality in a prospective follow-up of 89 patients with severe, nonspecific valvular heart disease. PATIENTS AND METHODS: Between November 1, 1993, and March 31, 1994, 89 patients with valvular heart disease were assessed for the presence of anticardiolipin antibodies and lupus anticoagulant. The primary end point was thromboembolic events, and the secondary end points were cardiovascular mortality and overall mortality. RESULTS: All patients were followed up for a mean of 59 months; 1 patient (without aPLs) was lost to follow-up. Nineteen patients had increased titers of aPLs. Thromboembolic events were significantly more frequent in the aPL-positive group than in the aPL negative group (7/19 [37%] vs 8/70 [11%]; P=.01). Cardiovascular mortality tended to be higher in the aPL-positive group than in the aPL-negative group (3 [16%] vs 6 [9%]; P = .40). However, in multivariate Cox analysis, presence of aPLs was not an independent risk factor for thromboembolic events. CONCLUSION: Our results suggest that patients with severe valvular heart disease and aPLs have an increased risk for developing thromboembolic events. PMID- 12630583 TI - 2-Chlorodeoxyadenosine therapy for disseminated Langerhans cell histiocytosis. AB - OBJECTIVE: To evaluate the efficacy of 2-chlorodeoxyadenosine (2-CDA), a purine nucleoside analogue, in treating disseminated Langerhans cell histiocytosis (LCH). PATIENTS AND METHODS: We retrospectively reviewed the clinical records of 5 patients who were seen at our institution for histologically confirmed disseminated LCH, including 1 patient with central nervous system parenchymal involvement. These patients were treated consecutively with 2-CDA chemotherapy between December 1994 and January 2001. The patients ranged in age from 19 to 81 years, and the median pretreatment duration of disease was 23 months. Median follow-up after initiation of 2-CDA treatment was 33 months. 2 Chlorodeoxyadenosine was used as frontline therapy for 1 patient and as salvage therapy for the other patients. Patients generally received 0.7 mg/kg over 5 or 7 days; the median number of courses was 4. RESULTS: Complete responses were achieved in 3 patients, including the patient with central nervous system disease, which, to our knowledge, has not been described previously. Two other patients achieved partial responses. The overall response rate was 100%. Toxic effects consisted mainly of myelosuppression; 1 patient developed dermatomal herpes zoster infection. CONCLUSION: Our experience confirms the reported efficacy of 2-CDA in the treatment of LCH; however, the optimal timing and schedule of therapy remain to be determined. PMID- 12630585 TI - Chlamydia pneumoniae and coronary artery disease: the antibiotic trials. AB - Parallel with the mounting evidence that atherosclerosis has a major inflammatory component, provoking agents that may initiate and drive this process have been sought. Infectious agents such as Chlamydia pneumoniae have been alleged to be activators of inflammation that may contribute to atherosclerosis and thus coronary artery disease (CAD) and its associated complications. A logical pneumoniae extension of this theory whether treating C pneumoniae infection with antibiotics and/or modulating inflammatory processes can affect CAD and its sequelae. This article discusses the potential role of C pneumoniae in atherosclerosis, its detection, and the rationale for antibiotics. Additionally, it summarizes the current randomized clinical trials of antichlamydial antibiotics in patients with CAD and draws conclusions based on the results. PMID- 12630584 TI - Primer on medical genomics. Part VI: Genomics and molecular genetics in clinical practice. AB - An important milestone in medical science is the recent completion of a "working draft" of the human genome sequence. The identification of all human genes and their regulatory regions provides the framework to expedite our understanding of the molecular basis of disease. This advance has also formed the foundation for a broad range of genomic tools that can be applied to medical science. These developments in global gene and gene product analysis as well as targeted molecular genetic testing are destined to change the practice of modern medicine. Despite these exciting advances, many practicing clinicians perceive that the role of molecular genetics, especially that of genomics, is confined primarily to the research arena with little current clinical applicability. The aim of this article is to highlight advances in DNA/RNA-based methods of susceptibility screening, disease diagnosis and prognostication, and prediction of treatment outcome in regard to both drug toxicity and response as they apply to various areas of clinical medicine. PMID- 12630586 TI - Therapeutic role of angiotensin II receptor blockers in the treatment of heart failure. AB - Angiotensin II type 1 receptor blockers (ARBs) are generally as effective as angiotensin-converting enzyme (ACE) inhibitors in patients with hypertension. However, inhibition of angiotensin is not achieved completely through the blocking effects of ACE inhibitors, and the possibility of a non-ACE pathway for generation of angiotensin II has important implications for treating cardiovascular disease. The selective quality of ARBs for the angiotensin II type 1 (AT1) receptor may confer an advantage. In a recently reported trial, the ARB valsartan substantially improved patients' New York Heart Association class, clinical signs and symptoms, and quality of life and provided morbidity and mortality benefits in selected patients. Valsartan was recently approved to treat heart failure in patients who cannot be maintained on an ACE inhibitor. As a class, ARBs are well tolerated and have a good safety profile. PMID- 12630587 TI - 44-year-old woman with fatigue and dyspnea. PMID- 12630588 TI - Paroxysmal sleep as a presenting symptom of bilateral paramedian thalamic infarctions. AB - Bilateral paramedian thalamic infarctions may result in hypersomnolence, supranuclear palsy, and an amnestic syndrome. We describe an 82-year-old man who presented with recurrent, transient episodes of unresponsiveness secondary to bilateral paramedian thalamic infarctions. Electroencephalography was performed during one of these paroxysmal events and showed an abrupt transition to a stage 2 sleep pattern with spindle formation. PMID- 12630589 TI - Extragenital adenomatoid tumor of a mediastinal lymph node. AB - Adenomatoid tumors are benign neoplasms found predominantly in male and female genital tracts. Rare extragenital adenomatoid tumors have been discovered that involve serosal surfaces and nonmesothelial-lined organs such as adrenal glands. Since the discovery of adenomatoid tumors, their histogenetic origin has been debated. Many researchers support a mesothelial cell origin for adenomatoid tumors because these tumors characteristically express a mesothelial phenotype. Tumor derivation from primitive pluripotent mesenchymal cells and coelomic epithelium also has been suggested because of the anatomical distribution of the tumors. Despite their characteristic mesothelial phenotype and histological appearance, adenomatoid tumors have an extensive differential diagnosis that includes vascular neoplasms, malignant mesothelioma, germ cell tumors, and metastatic adenocarcinoma. Recognition of these tumors may be especially difficult when examined at frozen section and when adenomatoid tumors are encountered in rare extragenital sites. We describe an adenomatoid tumor of a mediastinal lymph node that was found incidentally during a redo Collis-Nissen gastroplasty. On frozen section examination, this tumor was misinterpreted as metastatic adenocarcinoma. The hematoxylin-eosin histological, immunohistochemical, and ultrastructural studies confirmed the mesothelial phenotype of this tumor. To our knowledge, this is the first description of a lymph node adenomatoid tumor. PMID- 12630590 TI - Therapeutics in pediatric epilepsy, Part 1: The new antiepileptic drugs and the ketogenic diet. AB - Epilepsy is one of the most common and challenging neurologic disorders affecting children. Although various modalities exist to treat pediatric-onset seizures, seizures in 25% of children who are diagnosed as having epilepsy remain refractory to available therapies. Of the 8 new antiepileptic drugs (AEDs) (felbamate, gabapentin, lamotrigine, topiramate, tiagabine, levetiracetam, oxcarbazepine, and zonisamide), all but 2 (zonisamide and levetiracetam) have received Food and Drug Administration approval for adjunctive use in the pediatric population. However, most of the new AEDs used in adults have also been used in children, beyond the AEDs' approved indications. The ultimate goal of patient management is to choose the therapeutic option that provides the best chance of improving the patient's quality of life. Issues that relate to treatment choice include the likelihood of seizure recurrence, type and severity of seizures, available AED efficacies and toxicities, need for hematologic monitoring, ease of dosing, underlying medical conditions, medication interactions, urgency of initiating therapy, and cost. In this review, we discuss these issues for each of the 8 new AEDs; we also discuss the ketogenic diet and briefly review the older AEDs. Knowledge of the available AEDs will enable the practitioner to choose the best drug or drugs for individual patients. PMID- 12630591 TI - Therapeutics in pediatric epilepsy, Part 2: Epilepsy surgery and vagus nerve stimulation. AB - When antiepileptic drugs fail to relieve seizures adequately in children and adolescents, more invasive therapies such as epilepsy surgery and an implanted device to stimulate the vagus nerve should be considered. Temporal lobectomy is an effective treatment of complex partial and secondarily generalized tonic clonic seizures arising in the mesial structures or lateral temporal neocortex. Excellent outcomes (seizure free or rare, nondisabling seizures) are achieved in at least 70% of children. The most common adverse effect is a superior quadrant field cut that is usually asymptomatic. Transient and more long-lasting language difficulties have been reported when the surgery involves the dominant temporal lobe. The excellent outcome rate for extratemporal surgery ranges from approximately 20% to 80%, with better results seen in patients with an identifiable lesion. Potential morbidity is related to the region of resected neocortex. Corpus callosotomy is an excellent procedure for palliation but is not a cure for seizures that cause falls, with substantial improvement seen in more than 80% of patients. Potential adverse effects include more intense focal seizures and dysphasia, depending on the developmental level of the individual. Hemispherectomy provides seizure relief in 60% to 80% of patients with hemispherical pathologies such as Sturge-Weber or Rasmussen syndromes. Operative mortality has been reported in the range of 0% to 6%; other morbidities include infection and hydrocephalus. Stimulation of the vagus nerve has reduced partial seizures by 50% or more in approximately one third of patients. No adverse cognitive or systemic effects are associated with use of the implanted vagus nerve stimulator. PMID- 12630592 TI - Low-dose hormone replacement therapy for hot flashes. PMID- 12630593 TI - Determining optimal statin dosage. PMID- 12630594 TI - Atypical antipsychotic drugs for schizophrenia. PMID- 12630595 TI - HIV and AIDS among men of color who have sex with men and men of color who have sex with men and women: an epidemiological profile. PMID- 12630596 TI - HIV risk and prevention among Asian/Pacific Islander men who have sex with men: listen to our stories. AB - Despite increasing need for HIV prevention research and intervention programs, the voices and stories of Asian and Pacific Islander men who have sex with men (API MSM) have remained absent from HIV prevention literature. Five focus groups with API MSM (N = 38) were conducted to identify psychological, social, and cultural factors related to HIV risk and protection. Six themes were identified based on focus group discussion: (a) dual-identity status, (b) coming out and disclosure issues, (c) relationships and dating, (d) substance use, (e) sexual risk reduction strategies, and (f) health and social services. Narrative data indicate that multilevel HIV prevention intervention strategies are necessary for addressing the unique psychosocial and behavioral HIV risk factors among API MSM, such as dual stigma stemming from homophobia and racism, discomfort with sexuality, power dynamics and stereotypes in relationships with White men, substance use, and low utilization of health and social services. PMID- 12630597 TI - A model for understanding sexual health among Asian American/Pacific Islander men who have sex with men (MSM) in the United States. AB - The model to understand sexual health among Asian American/Pacific Islander men who have sex with men first locates the dynamic process in the home country, with its prevailing cultural norms including sexual mores, shame or stigma, sexual attitudes, sexual behavior, and drug use/abuse. Second, these cultural norms are modified by the migration/immigration experience. Third, these norms, beliefs, and practices are continually influenced by the process of acculturation as these men try to adjust to life in the United States. The effects of the first two domains may vary by the degree to which a particular immigrant community remains socially and culturally insulated from the mainstream community. Conceivably, the effect of home country and migration/immigration would be less significant for those who were either very young at the time of immigration or are born in the United States. PMID- 12630598 TI - The effect of partner characteristics on HIV infection among African American men who have sex with men in the Young Men's Survey, Los Angeles, 1999-2000. AB - Previous studies have documented disparities in HIV prevalence by race among men who have sex with men (MSM), even after adjusting for traditional risk factors. In this analysis of data collected for the 1999-2000 Los Angeles Young Men's Survey, a cross-sectional venue-based survey of MSM aged 23-29, we investigated whether information on male sex-partner characteristics accounts for some of the racial/ethnic differences in HIV prevalence. In this sample of survey participants, we observed that African American MSM reported similar or lower levels of HIV risk behaviors compared with White MSM but much higher HIV prevalence (26% vs. 7.4%, respectively). In an unadjusted logistic regression model, African American participants had 4.4 times higher odds of HIV infection compared with White participants. In a multiple logistic regression model adjusting for participant behaviors, we observed elevation of the relative odds of HIV infection for African Americans compared with Whites (odds ratio [OR] = 6.9, 95% confidence limits [CL] = 2.5, 19). In a fully adjusted model, controlling for the effects of having older partners and more African American partners, we observed a 20% reduction in the relative odds of HIV for African American participants compared with White participants (OR = 5.5, 95% CL = 1.8, 17). Our findings suggest that differences in male partner types, namely older and African American partners, may account for some of the observed racial disparity in HIV infection, especially for African American MSM compared with White MSM in Los Angeles. PMID- 12630599 TI - Age and race mixing patterns of sexual partnerships among Asian men who have sex with men: implications for HIV transmission and prevention. AB - Recent studies of U.S. men who have sex with men (MSM) have reported high rates of unprotected anal intercourse but low rates of HIV infection among Asian and Pacific Islander (API) men. We investigated this discrepancy by examining the characteristics of sexual partnerships among API MSM. Multivariate logistic regression analyses found that having an API partner was related to having unprotected anal intercourse after controlling for respondent's age, number of sexual partners, and partner type. Having an API partner and a younger partner were related to respondent reports of unprotected insertive anal intercourse. However, a partner's race and age did not predict unprotected receptive anal intercourse. Study findings suggest that HIV prevalence among API MSM may have remained relatively low because higher risk sexual practices occur more frequently within a lower risk API group compared with higher risk non-API groups. However, because the possibility for the rise of HV infection in this population still exists if HIV spreads from older to younger men, HIV prevention efforts must target partner characteristics to maintain low HIV prevalence among API MSM. PMID- 12630600 TI - Psychosocial predictors of risky sexual behaviors in African American men: implications for prevention. AB - Psychosocial predictors of sexual risk taking were investigated in a community sample of 502 HIV-positive and HIV-negative African American men enrolled in the African American Health Project. Poisson regression analyses were used to estimate the relative contributions of psychosocial risk and protective factors in predicting sexual risk as measured by a sexual risk behavior index. HIV negative men, men who have sex with men and women (MSM/W), and men who have sex with men (MSM) engaged in more high-risk sexual behaviors than heterosexuals and HIV-positive men, but men who were HIV-positive carried a heavier burden of psychosocial risk factors. High psychological distress, being HIV-negative, older age, low socioeconomic status (SES), and being an MSM/W were the best predictors of sexual risk. HIV serostatus and sexual orientation differences were obtained, with high psychological distress being the most consistent predictor regardless of serostatus or sexual orientation. Results confirm previous findings of riskier sexual lifestyle among MSM/W, men with low SES, and men who are experiencing significant psychological distress. PMID- 12630601 TI - Risk among young gay and bisexual men living with HIV. AB - The influence of sexual orientation was examined on sexual risk behaviors, disclosure patterns, substance use, and stressful live events among 231 gay and bisexual youth living with HIV. Youth were mainly of ethnic minority heritage: 69.3% were Hispanic, African American, or biracial. Although there were significant differences in gay and bisexual youth's self-label and patterns of sexual attraction and sexual partners, there were few differences in other behavioral risk acts. The frequency of sexual risk acts, substance use, stressful life events, and child sexual abuse were similar for gay and bisexual youth and similar across ethnic groups. Disclosure of sexual orientation was significantly more common among gay youth compared with bisexual youth. Gay and bisexual identification appears to be a more critical factor than ethnicity in placing youth at risk for HIV. Care providers need to screen youth for sexual orientation and behaviors at a young age and inquire about age of partners, substance use, and history of sexual abuse. PMID- 12630602 TI - Latinos' community involvement in HIV/AIDS: organizational and individual perspectives on volunteering. AB - Community involvement (e.g., volunteerism, activism) in HIV/AIDS may be an effective prevention strategy. Through involvement in HIV/AIDS-related organizations, individuals may develop a positive sense of themselves, maintain HIV preventive behaviors, and create community change. In this paper we examine the types of activities, motives, consequences, and deterrents to community involvement among Latino gay men using both community organizations' and Latino gay men's perspectives. Data come from an exploratory study in Chicago. It included telephone interviews with HIV/AIDS organizations (N = 62) and in-depth interviews with Latino gay men (n = 6 volunteers; n = 7 no volunteers). We found that organizations have few Latino volunteers and that the deterrents to involvement are stigma of HIV/AIDS and homosexuality, racism, and apathy. Among the positive consequences, we found an increase in self-esteem, sense of empowerment, and safer sex behaviors. PMID- 12630603 TI - Repeat HIV testing among Hispanic men who have sex with men--a sign of risk, prevention, or reassurance? AB - This study examined factors associated with repeat (n > or = 3 lifetime) and regular (n > or = 2 times per year, for a minimum of 1 year) HIV testing among a community sample of 538 seronegative Hispanic men who have sex with men (MSM). Bilingual staff interviewed respondents anonymously at public venues in South Florida. We compared (a) repeat testers with nonrepeat testers and (b) regular testers with nonregular testers. Results of logistic regression analyses indicated that repeat testers were more likely to be older, more educated, have a history of sexually transmitted disease, and have more sex partners than nonrepeat testers. Regular testers were more likely to be younger, have lower HIV risk perceptions, and have intentionally taken their first HIV test than were nonregular testers. They were also more likely to engage in oral sex and to only engage in 100% protected insertive anal sex. These findings suggest the importance of studying both the frequency and regularity of HIV testing behaviors, and using them to design interventions to promote testing among Hispanic MSM who are most at risk. PMID- 12630604 TI - Sexually transmitted diseases surveillance among incarcerated men who have sex with men--an opportunity for HIV prevention. AB - There is an urgent need for behavioral surveillance and prevention strategies specifically designed for men who have sex with men (MSM) who exhibit high-risk behavior. This report describes a sentinel screening program designed to identify sexually transmitted infections among incarcerated MSM. Approximately 2,200 inmates were screened for HIV, syphilis, gonorrhea, and chlamydia from a segregated unit of men who have sex with men in Los Angeles County Men's Central Jail. A convenience sample of 217 participants was offered a survey to assess risk behaviors. Screening activities yielded a 12.4% HIV, 3.1% chlamydia, 1.5% gonorrhea, and 0.3% early syphilis prevalence. The high HIV prevalence found in this population indicates the opportunity for HIV prevention within sexually transmitted disease (STD) screening of incarcerated MSM. Bridging HIV prevention into STD screening will require the integration of primary and secondary prevention services within the jail including case management, early treatment, and referral services. PMID- 12630605 TI - Adapting an evidence-based intervention: tales of the Hustler Project. AB - The field of HIV prevention is rapidly moving toward evidence-based practice as the standard way community-based organizations should do business. Current discussion about how to transfer evidence-based HIV prevention programs and practices to community-based organizations favors the perspectives of those scientists who hope to disseminate their programs to service providers. To date, there are few descriptions of community-based organizations' experiences adopting programs presented in the HIV prevention literature. Understanding and learning from community-based organizations' experiences of adopting programs can inform intervention design and the theory and practice of technology transfer. The current article describes one organization's experience adopting the popular opinion leader intervention. Implications for technology transfer are provided. PMID- 12630606 TI - A preliminary study of the optimal anesthesia positioning for the morbidly obese patient. AB - BACKGROUND: Hypoxemia during the induction of general anesthesia for the morbidly obese patient is a major concern of anesthesiologists. The etiology of this pathophysiological problem is multifactorial, and patient positioning may be a contributing factor. The present study was designed to identify optimal patient positioning for the induction of general anesthesia that minimizes the risk of hypoxemia in these patients. METHODS: 26 morbidly obese patients (body mass index BMI 56 +/- 3) were randomly assigned to one of three positions for induction of anesthesia: 1) 30 degrees Reverse Trendelenburg; 2) Supine-Horizontal; 3) 30 degrees Back Up Fowler. Mask ventilation, full neuromuscular paralysis and direct laryngoscopy were performed. Any airway difficulties were noted. After endotracheal tube placement, subjects were ventilated for 5 minutes with 1% isoflurane in a mixture of 50% oxygen/50% air and then disconnected from the ventilation circuit. The time required for capillary oxygen saturation (SaO2), as measured by pulse oximeter, to decline from 100% to 92% was noted and identified as the safe apnea period (SAP). Ventilation was then immediately re-established. The lowest SaO2 after resuming ventilation and the time from that nadir to an SaO2 of 97% were also recorded. RESULTS: BMI and hip-waist ratios of patients in groups 1, 2 and 3 did not significantly differ. There were no differences in airway difficulties between the different groups. The SAP in groups 1, 2 and 3 was 178 +/- 55, 123 +/- 24 and 153 +/- 63 seconds, respectively. The SaO2 of patients in the reverse Trendelenburg position dropped the least and took the shortest time to recover to 97%. CONCLUSIONS: In morbidly obese patients, the 30 degrees Reverse Trendelenburg position provided the longest SAP when compared to the 30 degrees Back Up Fowler and Horizontal-Supine positions. Since on induction of general anesthesia morbidly obese patients may be difficult to mask ventilate and/or intubate, this extra time may preclude adverse sequelae resulting from hypoxemia. Therefore, Reverse Trendelenburg is recommended as the optimal position for induction. PMID- 12630607 TI - The Magenstrasse and Mill operation for morbid obesity. AB - BACKGROUND: Our aim was to evolve a simpler, more physiological type of gastroplasty that would dispense with implanted foreign material such as bands and reservoirs. The Magenstrasse, or "street of the stomach", is a long narrow tube fashioned from the lesser curvature, which conveys food from the esophagus to the antral Mill. Normal antral grinding of solid food and antro-pyloro duodenal regulation of gastric emptying and secretion are preserved. METHODS: 100 patients with morbid obesity (83M, 17F, mean age 40 years) were treated by the Magenstrasse and Mill procedure and followed-up for 1-5 years. Mean preoperative BMI was 46.3 kg/m2, and mean excess weight was 106%. RESULTS: Operative mortality was 0. Major complications occurred in 4% of patients. There were few side effects, although mild heartburn was fairly common. Mean weight loss was 38 kg (+/- 14 kg), equivalent to 60% of excess weight, achieved within 1 year of operation, after which no further significant gain or loss of weight occurred. CONCLUSIONS: The Magenstrasse and Mill procedure is the simplest and most physiological gastroplasty yet described. Many of the drawbacks of vertical banded gastroplasty, adjustable banding and gastric bypass are avoided. It is safe, has few side-effects and leads to major and durable weight losses, similar to those produced by other types of gastroplasty. PMID- 12630609 TI - Obese patients have inadequate protein intake related to protein intolerance up to 1 year following Roux-en-Y gastric bypass. AB - BACKGROUND: Inadequate protein intake is a concern following Roux-en-Y gastric bypass (RYGBP). The small gastric pouch and bypass restrict energy intake and may lead to insufficient protein intake and absorption, and excess loss of lean tissue. METHODS: We evaluated protein intake in 93 (77 F, 16 M) morbidly obese individuals (BMI = 52.0 +/- 12.9 [SD]) who underwent RYGBP at our medical center. Participants completed 24-hr food recalls and received nutritional counseling at 3, 6, and 12 months following surgery. RESULTS: Daily energy intake (kcal/day) increased from 849 +/- 329 (SD) at 3 months to 1,101 +/- 400 at 12 months (P = .009). Protein intake also increased (g/day) from 45.6 +/- 14.2 at 3 months to 58.5 +/- 17.1 at 12 months (P = .04), and as a percentage of goal protein intake from 55.1% +/- 23.0 at 3 months to 73.5% +/- 38.0 at 12 months (P = .02). Although energy and protein intake increased significantly over the 12-month period, protein intake at 12 months remained significantly lower (P = .01) than the daily recommended guidelines (1.5 g/kg IBW) for a low-energy restrictive diet. Energy intake did not differ in those who reported food intolerances at 3 months (P = .77) or 6 months (P = .65), but was lower in them at 12 months (trend, P = .06). Also at 12 months, protein intake (P = .02) and percentage of protein intake goal (P = .04) were significantly lower in those with protein intolerance. CONCLUSIONS: These results suggest that postoperative patients consume insufficient amounts of protein, possibly mediated by protein intolerance. Protein supplementation following RYGBP deserves further consideration. PMID- 12630608 TI - Ghrelin: a gut-brain hormone: effect of gastric bypass surgery. AB - BACKGROUND: Ghrelin is a newly recognized gastric hormone with orexigenic and adipogenic properties, produced primarily by the stomach. Ghrelin is reduced in obesity. Weight loss is associated with an increase in fasting plasma ghrelin. We assessed the effect of massive weight loss on plasma ghrelin concentrations and its correlation with serum leptin levels and the presence of type 2 diabetes mellitus (DM) in severely obese patients. METHODS: A prospective study was conducted on 28 morbidly obese women (BMI 56.3 +/- 10.2 kg/m2) who underwent gastric bypass, divided into 2 groups: 14 non-diabetics (NGT) and 14 type 2 diabetics (DM2). Ghrelin and leptin were evaluated before silastic ring transected vertical gastric bypass, and again 12 months postoperatively. RESULTS: Fasting plasma ghrelin concentrations were 56% lower in NGT and 59% lower in DM2 compared with a lean control group (P < 0.001). There was no difference in ghrelin levels between NGT and DM2 groups before and after surgery (P > 0.05). Ghrelin was negatively correlated with leptin before gastric bypass surgery (r = 0.51, P < 0.01). The mean plasma ghrelin concentration decreased significantly after surgery in both groups (P < 0.001). CONCLUSION: Ghrelin was inversely related to leptin concentrations. Presence of diabetes did not affect the ghrelin pattern. Reduced production of ghrelin after gastric bypass could be partly responsible for the lack of hyperphagia and thus for the weight loss. PMID- 12630610 TI - Analysis of cost outliers after gastric bypass surgery: what can we learn? AB - BACKGROUND: A clinical pathway for gastric bypass surgery (GBS) implemented at our institution in 1999 resulted in reduced costs and decreased variability in patient care. However, a reanalysis of GBS hospital costs identified a 16% incidence of "cost outliers". We hypothesized that analysis of clinical variables would identify factors associated with increased hospital costs following GBS. METHODS: Medical records and financial data for 91 GBS patients from November 2000 to July 2001 were reviewed. Patients with costs >1 SD above the total hospital cost mean comprised the cost outlier (CO) group, while the remaining patients were considered the normal cost (NC) group. Potential etiologies for COs included patient demographics, the number and severity of medical co-morbidities, surgical factors, and major postoperative complications. RESULTS: There were 15 patients in the CO group, and 76 patients in the NC group. Patient demographics were similar in both groups. Diabetes mellitus and severe medical co-morbidities, especially sleep apnea and degenerative joint disease were more common in the CO group (60% vs 9.2%, P < 0.05 vs NC). The incidence of major complications (33% vs 8%) was significantly increased in the CO group (P < 0.05 vs NC). CONCLUSIONS: Despite utilization of a clinical pathway for GBS, 16% of patients were "cost outliers". Factors associated with increased hospital costs after GBS included severe medical co-morbidities (especially diabetes mellitus and sleep apnea) and the occurrence of major postoperative complications. Prospective identification of "high risk" GBS patients may allow hospitals with bariatric surgery programs to modify perioperative care and eliminate potential cost outliers. PMID- 12630611 TI - Reinforcing gastric staple-lines with bovine pericardial strips may decrease the likelihood of gastric leak after laparoscopic Roux-en-Y gastric bypass. AB - BACKGROUND: Staple-line leakage is a potentially devastating complication of Roux en-Y gastric bypass (RYGBP). Bovine pericardial strips (BPS) have been used to reinforce staple-lines in pulmonary resections and have been shown to decrease air-leaks. This study examined the use of BPS to decrease gastric staple-line leaks. METHODS: 250 consecutive patients undergoing laparoscopic RYGBP had BPS used for the gastric pouch. Ease of use, operative complications, and visual bleeding were recorded. In addition, the postoperative leak rate was compared to 100 consecutive patients operated on prior to the study when BPS was not used. RESULTS: Patient characteristics were the same for both groups and comparable with most published data. With the use of BPS, there were no operative complications and no meaningful increase in operating time. BPS reinforced staple lines had no visual bleeding. In the 100 cases without BPS, there were 2 staple line leaks (2%) both of which required emergency exploration. One additional patient was subsequently found to have a gastrogastric fistula. In the 250 patients who had BPS-reinforced staple-lines, there were no acute leaks. Three patients were subsequently found to have gastrogastric fistula. None required emergency surgery. CONCLUSION: In this non-randomized trial, BPS were found to be easy and safe to use. In addition, staple-line hemorrhage was essentially non existent. Although the reduction in gastric staple-line leak rate may also be attributed to learning curve, there were no acute leaks in 250 patients with BPS, which is below the published norms. PMID- 12630612 TI - Fibrin glue as a sealant for high-risk anastomosis in surgery for morbid obesity. AB - BACKGROUND: Fibrin sealants promote hemostasis and wound healing. Complex revisional surgery is performed for morbid obesity, and high-risk patients undergo weight loss surgery routinely. Fibrin sealant, Tisseel, was used by one surgeon on 120 consecutive patients at the gastrojejunal anastomosis in Roux-en-Y gastric bypass (RYGBP). We hypothesized that the application of fibrin sealant would decrease anastomotic leaks. METHODS: One surgeon (Surgeon A) used fibrin sealant for 120 consecutive patients, while two other surgeons (Surgeons B & C) served as controls and did not use fibrin glue for their last 120 patients. Surgeon A did not use fibrin glue in 120 patients to serve as an internal control. All 480 patients underwent a RYGBP. Fibrin glue was applied at the gastrojejunal anastomosis. RESULTS: The fibrin sealant group did not have any documented leaks on the previous 120 patients, while 5 patients with Surgeon B, 2 patients with Surgeon C and 1 patient with Surgeon A without fibrin sealant experienced enteric leaks requiring re-operation, drainage, or long-term total parenteral nutrition (N = 480 total patients). CONCLUSIONS: Fibrin sealant may be useful in preventing leaks and promoting healing of the "high risk" anastomosis during complex gastrointestinal surgery. While the cost of fibrin glue is to be considered, re-operation and management of subsequent enterocutanous fistulas or anastomotic strictures may be more costly than routine use for high-risk morbidly obese patients. PMID- 12630613 TI - Dysfunctional immune-privilege in morbid obesity: implications and effect of gastric bypass surgery. AB - BACKGROUND: Despite the epidemiological evidence linking obesity and cancer, there has never been a causal link. We believe the chronic inflammation present in obesity may predispose the obese to cancer through Fas-receptor over expression and L-selectin under-expression in leukocytes, and elevated Fas ligand secretion in tumors affecting the morbidly obese. METHODS: Leukocytes from 25 patients having gastric bypass surgery were compared to 15 normal controls preoperatively and at 1, 3, 6, and 12 months postoperatively using flow cytometry to measure CD3, CD4, CD8, CD56, CD62 (L-selectin), CD69, and CD95 (Fas antigen) expression on T lymphocytes, B lymphocytes, natural killer cells, and neutrophils. RESULTS: The percentage of CD95 + T cells was significantly elevated from controls (69.4% vs 56%, P = 0.005). This difference persisted through 1 month postoperatively. Furthermore, expression of CD95 per cell, was significantly greater in these patients than that of the controls (80.2 vs 62.6 gmf, P = 0.018) preoperatively, and this continued to 1 month. Polymorphonuclear cells also displayed a similar elevation in CD95 gmf expression preoperatively (54.1 vs 40.7 gmf, P = 0.023) which normalized by 3 months. Natural killer cells did not display elevated numbers of CD95 gmf preoperatively, but they did experience a significant decline by 12 months. Additionally, there was significant reduction in the number of naiveT cells [(T cells without L-selectin (CD62L)], when compared to normals preoperatively (41.8% vs 51.3%, P = 0.001). There was no statistical difference between the postoperative patients and the controls by 3 months. CD69 was not different at baseline from controls in T or B cells, but there was a significant decrease by 12 months. CONCLUSION: The reduced expression of L-selectin combined with the elevated levels of CD95 suggests that morbid obesity predisposes patients to sites of immune privilege. This could be the mechanism for increased rates of cancer and wound infections seen in obesity. Surgically-induced weight loss eliminates these risk factors. PMID- 12630614 TI - Gastric bypass is an effective treatment for obstructive sleep apnea in patients with clinically significant obesity. AB - BACKGROUND: We have demonstrated that obstructive sleep apnea (OSA) is prevalent in 60% of patients undergoing bariatric surgery. A study was conducted to determine whether weight loss following bariatric surgery ameliorates OSA. METHODS: All 100 consecutive patients with symptoms of OSA were prospectively evaluated by polysomnography before gastric bypass. Preoperative and postoperative scores of Epworth Sleepiness Scale (ESS), Respiratory Disturbance Index (RDI), and other parameters of sleep quality were compared using t-test. RESULTS: Preoperative RDI was 40 +/- 4 (normal 5 events/hour, n = 100). 13 patients had no OSA, 29 had mild OSA, while the remaining 58 patients were treated preoperatively for moderate-severe OSA. At a median of 6 months follow up, BMI and ESS scores improved (38 +/- 1 vs 54 +/- 1 kg/m2, 6 +/- 1 vs 12 +/- 0.1, P < 0.001, postoperatively vs preoperatively). To date, 11 patients have completed postoperative polysomnography (3-21 months) after losing weight (BMI 40 +/- 2 vs 62 +/- 3 kg/m2, P < 0.001). There was significant improvement in ESS (3 +/- 1 vs 14 +/- 2), minimum O2 saturation (SpO2 86 +/- 2 vs 77 +/- 5), sleep efficiency (85 +/- 2% vs 65 +/- 5%), all P < 0.001, postop vs preop; and RDI (56 +/- 13 vs 23 +/- 7, P = 0.041). Regression analysis demonstrated no correlation between preoperative BMI, ESS score and the severity of OSA; and no correlation between % excess body weight loss and postoperative RDI. CONCLUSION: Weight loss following gastric bypass results in profound improvement in OSA. The severity of apnea cannot be reliably predicted by preoperative BMI and ESS; therefore, patients with symptoms of OSA should undergo polysomnography. PMID- 12630615 TI - Early gastrointestinal hemorrhage after laparoscopic gastric bypass. AB - BACKGROUND: Early gastrointestinal (GI) hemorrhage after open gastric bypass has been infrequently reported. The aim of this study was to examine the incidence of early GI hemorrhage after laparoscopic Roux-en-Y gastric bypass (LRYGBP), its presentation, and possible treatment options. METHODS: A retrospective review of 5 patients who developed early postoperative GI hemorrhage after LRYGBP was performed. The charts were reviewed for demographics, clinical presentation, diagnostic evaluation, and treatment. All patients underwent a transected LRYGBP with creation of the gastrojejunostomy anastomosis with a circular stapler and the jejunojejunostomy anastomosis with a linear stapler. RESULTS: Of the 155 patients in our database who underwent LRYGBP, 5 (3.2%) developed early clinical GI hemorrhage. There were 2 males with an average age of 40 years. Clinical presentations of GI hemorrhage were hematemesis (2 patients), bright red blood per rectum (1 patient), melena (1 patient), and hypotension (1 patient). A diagnostic study (nuclear scintigraphy) was performed in only 1 of 5 patients. 3 of 5 patients were managed nonoperatively; 2 patients required fluid and blood resuscitation, and the other patient was managed without blood transfusion. The onset of hemorrhage in these 3 patients occurred 24 hours postoperatively or later. 2 of 5 patients required operative intervention for control of hemorrhage. The onset of hemorrhage or hypotension in these 2 patients occurred within 12 hours after surgery. The sites of hemorrhage were at the gastric remnant staple lines in 1 patient and at the gastrojejunostomy and gastric remnant staple-lines in the other patient. CONCLUSION: Early GI hemorrhage is a potential complication after transected LRYGBP. Early reoperative intervention should be performed for patients with hemodynamic instability and patients with early onset of hemorrhage after surgery. PMID- 12630616 TI - Routine upper gastrointestinal Gastrografin swallow after laparoscopic Roux-en-Y gastric bypass. AB - BACKGROUND: Upper gastrointestinal (UGI) swallow radiographs following laparoscopic Roux-en-Y gastric bypass (LRYGBP) may detect an obstruction or an anastomotic leak. The aim of our study was to determine the efficacy of routine imaging following LRYGBP. METHODS: Radiograph reports were reviewed for 201 consecutive LRYGBP operations between April 1999 and June 2001. UGI swallow used Gastrografin, static films, fluoroscopic video, and a delayed image at 10 minutes. Mean values with one standard deviation were tested for significance (P < 0.05) using the Mann-Whitney U test statistic. RESULTS: Of 198 available reports, UGI detected jejunal efferent (Roux) limb narrowing (n = 17), partial obstruction (n = 12), anastomotic leak (n = 3), complete bowel obstruction (n = 3), diverticulum (n = 1), hiatal hernia (n = 1), and proximal Roux limb narrowing (n = 1). A normal study was reported in 160 cases (81%). Partial obstruction resolved without intervention. Complete obstruction required re-operation. Compared to 6 patients who developed delayed leaks, early identification of a leak by routine UGI swallow resulted in a shorter hospital stay (mean 7.7 +/- 1.5 days vs 40.2 +/- 12.3 days, P < 0.03). CONCLUSIONS: Early intervention after UGI swallow may lessen morbidity. Routine UGI swallow following LRYGBP does not obviate the importance of close clinical follow-up. PMID- 12630617 TI - Sensitivity and specificity of postoperative upper GI series following gastric bypass. AB - BACKGROUND: Routine postoperative GI series has been common before discharging gastric bypass patients. 78,000 operations were performed in the USA in 2002. At 75 dollars each, the total annual expenditure for the upper GI series approaches 6 million dollars. This study examines the value of performing routine upper GI series. MATERIALS AND METHODS: From 1996 to 2000, 396 open gastric bypass procedures were performed by one surgeon at the University Medical Center. 242 randomly selected charts were retrospectively reviewed for signs and symptoms possibly related to leak or obstruction. Radiology reports were compared with clinical findings. RESULTS: 82% of patients (192/242) were discharged following unremarkable postoperative courses and normal x-rays. 18% (44/242) exhibited one or more clinical signs suspicious of leak or obstruction. These included fever, tachycardia, tachypnea, inordinate pain, elevated white cell count or GI hemorrhage. Leak was reported in 5, and obstruction in 5. 4 patients with reported leaks were re-operated: 2 were positive for unconfined leak requiring surgical treatment; 2 had negative laparotomies. The 2 patients (0.82%) with free leakage had dramatic clinical deterioration, and x-rays were confirmatory rather than diagnostic. 1 patient with a minimal confined leak was treated non operatively. 8 films were misread as showing a leak when none was present: 2 underwent negative laparotomy, the others being correctly interpreted after review. 8 of 10 initial interpretations were falsely positive. CONCLUSION: Routine postoperative GI series following gastric bypass is not beneficial. All true leaks are demonstrated when x-rays are indicated. We recommend GI series only when clinically indicated. GI series had low positive predictive value for leak. PMID- 12630618 TI - Elective cholecystectomy during laparoscopic Roux-en-Y gastric bypass: is it worth the wait? AB - BACKGROUND: Combined gastric bypass and cholecystectomy have been advocated for open bariatric procedures. Our goal was to evaluate the safety of this technique in laparoscopic bariatric surgery patients with gallstones diagnosed preoperatively. METHODS: 94 out of 556 consecutive morbidly obese patients (16.9%) underwent laparoscopic gastric bypass with simultaneous cholecystectomy (LGBP/LC) for cholelithiasis. RESULTS: 328 patients (59%) had a concomitant secondary procedure, most commonly cholecystectomy (28.7%). Preoperative BMI was 48.6 +/- 6.9 kg/m2 for LGBP/LC patients and 48.8 +/- 7.3 kg/m2 (P = 0.85) for LGBP alone. 5 patients had preoperative biliary colic; the others were asymptomatic for cholelithiasis. Postoperatively, at a mean follow-up of 7.6 +/- 6.7 months, the percent excess weight loss (%EWL) was 46.1 +/- 0.25 for the combined procedure vs 50.2 +/- 63.0 (P = 0.55) for LGBP alone. There were no conversions to open procedures for the LC. Port placement for the LGBP was not altered for LC. None required intraoperative cholangiography. Operative time for the combined procedure was 293.4 +/- 79.8 minutes vs 244.8 +/- 77.2 minutes for LGBP alone (P < 0.0001). Length of stay for the combined procedure was 4.35 +/- 10.8 days vs 2.69 +/- 1.8 days for LGBP alone (P = 0.0069). There were no postoperative bile leaks or bile duct injuries. CONCLUSION: Concomitant LGBP/LC is safe and feasible without altering port placement. Combining these procedures significantly increases operative time and nearly doubles the hospital stay. PMID- 12630619 TI - Who should be doing laparoscopic bariatric surgery? AB - BACKGROUND: The training and credentialing of surgeons for laparoscopic bariatric surgery is controversial. We sought to determine if there is an association between surgeons' practice and choice of open or laparoscopic bariatric surgery. METHODS: Members of the ASBS were surveyed via email. Associations were tested with Cochran-Mantel-Haenszel or Pearson's chi-square. RESULTS: 104/472 members responded; 65% were in private practice; 47% did 1-5 operations/week, 48% offered open procedures only, and 76% undertook gastric bypass. Respondents believe that laparoscopic procedures: should mimic open ones (77%), are safe (63%), should be evaluated by clinical trials (48%), and that expertise in bariatric surgery is more important than laparoscopic experience. 75% believe that courses and preceptorships are important. Regarding laparoscopic operations, surgeons doing only open procedures believe that: 1) the ASBS should be the main credentialing body; 2) surgeons should do > 25 open before laparoscopic ones; and 3) clinical trials are needed (P < 0.02, all). Surgeons with laparoscopic training or practices believe that laparoscopic surgery is safe and effective (P < 0.002). Both laparoscopic and open surgeons believe bariatric surgeons should be the only surgeons doing laparoscopic bariatric procedures (P < 0.008). CONCLUSIONS: There is consensus that laparoscopic bariatric surgery should be undertaken only by surgeons with strong interest in bariatric surgery. Laparoscopic bariatric surgeons should incorporate lessons learned from open surgery. Both laparoscopic and open bariatric surgeons should seek added expertise via courses and preceptorships. The skepticism of surgeons with 'open' practices could be addressed by clinical trials. The ASBS should maintain its leadership position and foster emerging technologies. PMID- 12630620 TI - Early experience with computer-mediated flexible circular stapling technique for upper gastrointestinal anastomosis. AB - BACKGROUND: Creating the proximal anastomosis in laparoscopic biliopancreatic diversion with duodenal switch (LBPD-DS) and Roux-en-Y gastric bypass (LRYGBP) is a critical step in ensuring the success of the procedures. The aim of this study was to assess the safety and efficiency of performing this anastomosis using a flexible, computerized, circular stapling device. METHOD: We prospectively monitored the use of a newly FDA-approved stapling device (SurgASSIST, Power Medical Intervention) for the construction of the proximal anastomosis by a variety of approaches and reviewed the charts of 10 patients. RESULTS: We successfully constructed 9 out of 10 proximal anastomoses: 2 gastro-jejunostomies and 7 duodeno-ileostomies, without any signs of leakage. In 2 patients, the stapling technique involved a transpyloric instrumental passage; both were complicated by the difficulty to pass either the flexible scope or the anvil through the narrow pyloric lumen. In 5 patients, the anvil was placed directly through a duodenotomy and no technical problems were encountered. The median time for performing the proximal anastomosis was 19 minutes (range 9-55). There were no postoperative complications in any patients. CONCLUSIONS: Stapling using the SurgASSIST was feasible and safe for performing laparoscopic anastomoses in bariatric bypass procedures. A duodenotomy for direct placement of the anvil into the postpyloric region seems to be most feasible for duodenoileostomies, while transoral passage of the anvil can be recommended for gastro-jejunostomies. In its current form, we do not recommend transoral placement of the flexible shaft of the SurgASSIST device. Further clinical trials need to be performed for comparison with existing devices. PMID- 12630621 TI - Circumferential abdominoplasty for sequential treatment after morbid obesity. AB - BACKGROUND: The severely obese patient, after considerable loss of weight, has large remaining skin folds. Classic transverse abdominoplasties leave bulges in the flanks. This article presents an alternative procedure, circumferential abdominoplasty, which involves the perimeter of the abdomen. METHODS: Average age of the patients was 39.5 years, consisting of 10 females and two males. Incisions were made like those of the classic transverse abdominoplasty, but were extended dorsally without tension on the suture-line. RESULTS: Pre and postoperative aspects revealed the huge impact achieved after resection of the excess panniculus, with improvement of body contour, posture, ambulation, social and psychological integration, hygiene and sexual performance. In some patients, seromas, partial dehiscences, flap infection and atelectasis were immediate complications. Hypertrophic scars and some remaining skin folds were late complications. CONCLUSION: The major disadvantage of the circumferential abdominoplasty is the resulting scar. However, this procedure should be taken into consideration as an option to achieve a more harmonious body contour. Complications are not enough to contraindicate the surgery, because the patients preferred better social and professional integration, as well as behavioral improvement due to enhancement in their self-confidence. PMID- 12630622 TI - Laparoscopic gastric banding in morbidly obese adolescents. AB - BACKGROUND: 4% of adolescents in the U.S.A. are obese, 80% of whom will become obese adults. Obesity in adolescence is associated with increased mortality and morbidity in adulthood. Is laparoscopic adjustable silicone gastric banding a safe and effective method of weight loss in morbidly obese adolescents? METHODS: Since 1996, data has been prospectively collected on all patients undergoing laparoscopic adjustable gastric banding (LAGB) by a single surgeon. Patients are reviewed at 6 and 12 weeks following surgery, then at 3 monthly intervals. Weight loss is measured in absolute terms, reduction in body mass index (BMI) and as percentage of excess weight loss. RESULTS: 17 patients with a median age of 17 (12 to 19) years underwent LAGB. Median follow-up was 25 (12 to 46) months. 2 complications occurred, 1 slipped band and 1 leaking port. BMI fell from a preoperative median of 44.7 to 30.2 kg/m2 at 24 months following surgery, corresponding to a median loss of 35.6 kg or 59.3% of excess weight. 13 of 17 patients (76.5%) lost at least 50% of their excess weight, and 9 of 11 patients (81.8%) had a BMI < 35 kg/m2 at 24 months following surgery. CONCLUSION: LAGB is a safe and effective method of weight loss in morbidly obese adolescents, at least in the medium term. Its role in preventing obesity and obesity-related disease in adulthood remains to be determined as part of our long-term study. PMID- 12630623 TI - Psychosocial consequences of weight loss following gastric banding for morbid obesity. AB - BACKGROUND: This study was performed to determine what consequences surgery for morbid obesity has on weight loss, problems in eating behavior, quality of life, physical appearance and mental state. METHOD: After a minimum follow-up of > 8 months (median follow-up 21 months, range 8-48 months), a questionnaire concerning extent of, satisfaction with and consequences of weight loss was mailed to 250 morbidly obese patients after laparoscopic Swedish adjustable gastric banding (SAGB). In addition, the partner's opinion regarding the operation was evaluated as well as the consequences of weight loss for partnership and sexual relationship. RESULTS: 160 patients (64%) completed and returned the questionnaire. Most patients (87%) were happy with the extent of weight loss. Weight loss, however, was connected with negative consequences for the body such as flabby skin (53%), abdominal skin overhang (47%) and pendulous breasts (42%). Patients who were satisfied with their postoperative physical appearance showed significantly less weight loss than did patients who were unhappy with their appearance (38 vs 54 kg). Most of the partners (91%) believed that the decision for SAGB was right. An improvement in partnership was reported by more than half of the partners (59%), and an improved sexual relationship by 45%. CONCLUSION: Laparoscopic SAGB is an effective surgical treatment for morbid obesity. However, the consequences of excess and rapid weight loss for physical appearance are negative in many cases. Well-directed information about the consequences of excess weight loss before SAGB and the possibilities and limits of plastic surgery must be given preoperatively to offset high and often unrealistic expectations. PMID- 12630624 TI - Pouch dilatation and slippage after adjustable gastric banding: is it still an issue? AB - BACKGROUND: Laparoscopic adjustable gastric banding (LAGB) in France is currently the most common bariatric surgical procedure for the treatment of severe obesity; its most reported complication is band slippage and/or pouch dilatation, which usually requires reoperation. It is highly important to assess whether a change in the operation could improve these results. METHODS: From April 1995 to October 2001, 973 patients underwent LAGB in our institution. Since January 1999, our technique changed: the band was positioned according to the so called "pars flaccida technique", ie. around the gastric vessel instead of close to the gastric wall. 511 patients had been operated before this period, and 462 after. Other details in the technique did not change (dissection above the lesser sac, no posterior stitch, three anterior stitches), meaning that potential differences could not be related to a learning curve. RESULTS: Band slippage occurred in 27 patients of the first group during the first period of 34 months (5.2%), and 5 more afterwards (total 6.2%). Only 3 patients of the second group (0.6%) had a slippage during the same period of time. CONCLUSION: Although the problem of band slippage is not likely to be completely solved, changing the technique has made it possible to decrease the rate of this complication. The height or the shape of different types of band also remain under scrutiny. PMID- 12630625 TI - Laparoscopic management of lap-band slippage. AB - BACKGROUND: The Lap-Band is generally associated with a low morbidity rate. Although gastric slippage through the band remains a concern, the rate has diminished with new band placement techniques. METHODS: Between November 2000 and June 2002, 198 Lap-Band Systems were inserted in patients in the Houston, Texas, area. 4 of these patients, plus an additional patient whose Lap-Band had been inserted by another surgeon, developed slippages, and in each case the gastric slip was corrected and the band was salvaged and repositioned laparoscopically. To facilitate dissection and repositioning of the band in 3 of these patients, the band had to be unlocked using a simple laparoscopic technique. RESULTS: All patients were discharged the morning following surgery with no complications and good position of the band as evidenced by esophagogram study and resolution of symptoms. After 2 to 12 months follow-up, all 5 patients remain symptom-free and continue to lose weight. CONCLUSIONS: Gastric slippage with the Lap-Band can be managed by laparoscopic salvage and repositioning of the slipped band. PMID- 12630626 TI - Postoperative management of laparoscopic gastric banding. AB - BACKGROUND: The authors investigated the postoperative management of morbidly obese patients treated by laparoscopic adjustable gastric banding (LAGB) with the Lap-Band System. METHODS: The 3-year postoperative band management is presented in 379 morbidly obese patients, divided according to intra-operative band filling and quartiles of maximum postoperative band filling. RESULTS: LAGB resulted in a 40.8 +/- 24.5 percent excess weight loss (%EWL). Stoma stenosis occurred in 87 patients (23.0%), pouch dilatation in 52 (13.7%) and esophageal dilatation in 22 (5.8%). Most band-related complications were controlled by simple band deflation. The mean number of postoperative band adjustments was 2.3 +/- 1.7, and mean maximum band filling after surgery was 2.8 +/- 1.2 ml. Weight loss at 3 years was identical in 205 patients who had the band completely unfilled at surgery and in 174 patients who had the band filled with 1 to 3 ml of sterile saline. The rate of band-related complications was significantly lower in the first group. No differences in %EWL were observed between quartiles of maximum band filling after surgery. The rate of band-related complications increased with increasing levels of postoperative maximum band filling. In patients with the band filled with < 3.0 ml of sterile saline at 6 months, the inflation of further saline produced a dose-related increase in the rate of weight loss. In patients with the band filled with > 3.0 ml of sterile saline at 6 months, the inflation of further saline was associated with a reduced %EWL. CONCLUSION: Postoperative adjustability of the Lap-Band was useful in the treatment of band-related complications and was able to significantly influence the rate of weight loss. On the other hand, aggressive postoperative band filling was associated with an increased rate of complications. PMID- 12630627 TI - Access-port complications after laparoscopic gastric banding. AB - BACKGROUND: The aim of this retrospective study was to identify complications related to the access-port, after Lap-Band system placement by laparoscopy. METHODS: The records of 333 morbidly obese patients who underwent laparoscopic adjustable gastric banding (LAGB) were reviewed for the overall surgical complications. Data was further analyzed regarding the complications related to the access-port. RESULTS: From January 1999 to December 2001, the overall complication-rate with the LAGB was 25.8%. 45 complications (13.5%) were related to the access-port in 34 patients following LAGB placement. The 45 access-port complications were distributed as follows: infection 51.1%, tubing disconnection 17.7%, dislodgment of the access-port 15.6%, leak of the reservoir 11.1%, and skin ulceration by the port 4.45%. CONCLUSION: The integrity of the Lap-Band system is essential to achieve the objective of the operation: weight loss. Complications related to the access-port were relatively frequent, but preventable. PMID- 12630629 TI - Carcinoma in the gastric pouch years after vertical banded gastroplasty. AB - A 67-year-old lady presented with anemia and weight loss 15 years after vertical banded gastroplasty. The cancer was confined to the pouch, which is suggestive of a relationship to the anti-obesity surgery. A brief review with possible contributing factors is presented. PMID- 12630628 TI - Technical modification of laparoscopic vertical banded gastroplasty. AB - BACKGROUND: Technical improvements of laparoscopic bariatric procedures are important to minimize operative time and increase safety and simplicity. METHODS: A modification is described of the "classic" Mason-MacLean vertical banded gastroplasty (VBG), performed by laparoscopy, with wedge resection of the gastric fundus, thus avoiding the time-consuming and technically difficult gastro gastrostomy window. RESULTS: The technique used was simple and safe, and required less operative time than the "classic" method, without serious intra- or postoperative complications. Weight loss in 18 patients who underwent the modified laparoscopic VBG and were followed-up for 1 year was equal to that achieved in patients who underwent open VBG at our Institution. Gastro-gastric fistula was not observed in upper GI barium studies performed 12 months postoperatively. CONCLUSION: The modified technique is preferable when lap-VBG is indicated for surgical treatment of morbidly obese patients. PMID- 12630630 TI - Succinylcholine and morbid obesity. PMID- 12630631 TI - Laparoscopic gastric bypass with BMI > or = 60. PMID- 12630632 TI - Thromboembolism prophylaxis and cesarean section. PMID- 12630633 TI - Can intensive primary care prevent primary intensive care? PMID- 12630634 TI - Heart failure mortality among older Medicare beneficiaries: association with left ventricular function evaluation and angiotensin-converting enzyme inhibitor use. AB - BACKGROUND: Left ventricular function evaluation and angiotensin-converting enzyme (ACE) inhibitor use are the two basic indicators of heart failure quality of care. In this retrospective follow-up study, we analyzed the association between these two quality indicators and mortality in elderly hospitalized heart failure patients. METHODS: The patients in our study were older Alabama Medicare beneficiaries discharged with a diagnosis of heart failure in 1994. Cox regression analyses, adjusted for various patient and care characteristics, were performed to estimate the overall mortality rate. RESULTS: The mean age of the 1,090 patients in our study was 79+/-7.5 years. Both left ventricular function evaluation (hazard ratio, 0.83; 95% confidence interval, 0.705-0.976) and ACE inhibitor use (hazard ratio, 0.77; 95% confidence interval, 0.655-0.905) were associated with a lower 3-year mortality rate. Adjustment for various patient and care characteristics did not alter these associations. CONCLUSION: Left ventricular function evaluation and ACE inhibitor use were each associated with increased survival time in older Medicare beneficiaries with heart failure. PMID- 12630635 TI - Personal breast-feeding behaviors of female physicians in Mississippi. AB - BACKGROUND: In this study, we examined the personal breast-feeding behaviors of female physicians in Mississippi. METHOD: Two hundred fifteen of 350 female physicians responded to a survey inquiring of their personal breast-feeding behaviors. RESULTS: One hundred fifty-five mothers (74%) reported having biologic children, and 146 (94.2%) breast-fed at least 1 child. Approximately 21% of the responding mothers breast-fed their first-born children for at least 6 months. There was a positive relationship between the duration of breast-feeding of older children and the breast-feeding duration for younger children. The major reasons for weaning were return to work, diminishing milk supply, and lack of time to pump breast milk. CONCLUSION: The breast-feeding initiation rates among female physicians surpassed those of women in the general population, yet duration rates were comparable. Their own breast-feeding success might enhance the potential of female physicians as advocates and sources of credible information regarding breast-feeding; however, physicians need to be better educated regarding the management of breast-feeding. PMID- 12630636 TI - Percutaneous drainage for treatment of infected pancreatic pseudocysts. AB - BACKGROUND: Infection of pancreatic pseudocysts is a potentially fatal complication that must be treated immediately. Despite numerous published reports about percutaneous treatment, the effectiveness of percutaneous catheter drainage (PCD) of infected pancreatic pseudocysts is still under discussion. METHODS: In this study, 30 patients (17 women) with 30 infected pancreatic pseudocysts were administered local anesthesia and underwent PCD performed with the use of a single-step trocar technique with computed tomographic guidance. The patients' ages varied from 27 to 74 years (mean age, 45 yr). The etiology was acute pancreatitis in 18 patients, chronic pancreatitis in 11 patients, and surgical trauma in 1 patient. RESULTS: No complications related to the procedure occurred in our series. The success rate was 96% (29 of 30 patients), with no recurrence during follow-up, which ranged from 2 to 58 months (mean follow-up, 27.2 mo). One patient had unsuccessful PCD and was subsequently treated surgically. CONCLUSION: Our findings indicate that PCD is a safe and effective front-line treatment for patients with infected pancreatic pseudocysts. PMID- 12630637 TI - Anxiety and depression in hospitalized patients in resistant organism isolation. AB - BACKGROUND: Previous studies have reported an increase in psychiatric symptoms in seriously ill patients who were placed in resistant organism isolation. We conducted this study to assess whether there is an increase in symptoms of anxiety and depression in patients who are not critically ill and are placed in isolation. METHODS: Patients hospitalized with methicillin-resistant Staphylococcus aureus or vancomycin-resistant Enterococcus species infections were evaluated with the Hamilton Anxiety Rating Scale and the Hamilton Depression Rating Scale at baseline and again during hospitalization. The results were then compared with the results of patients who were hospitalized for infectious diseases that did not require isolation. RESULTS: Patients in isolation had significantly higher scores on both the anxiety and depression scales at the time of follow-up than did patients who were not isolated. There was no significant difference between the scores of the two groups before isolation. CONCLUSION: The results of this preliminary study suggest that placement in resistant organism isolation may increase hospitalized patients' levels of anxiety and depression. PMID- 12630638 TI - Thromboembolism prophylaxis and cesarean section: a survey of general obstetricians. AB - BACKGROUND: Venous thromboembolism (VTE) has historically remained a risk factor for obstetric patients, particularly those requiring cesarean section. The objective of this survey was to assess the response of general obstetricians to risk factor considerations for VTE prophylaxis and cesarean section. METHODS: A survey regarding VTE prophylaxis and cesarean section was sent to 113 obstetricians practicing in suburban Chicago and northern Wisconsin. RESULTS: There were 51 responses (45%) to the survey. Most respondents do not routinely use VTE prophylaxis for cesarean section in patients who may have one or more risk factors. CONCLUSION: There appears to be lack of consensus or appreciation for VTE and prophylaxis for cesarean section. Further studies and education are warranted. PMID- 12630639 TI - Access to health care professionals in rural Mississippi. AB - BACKGROUND: The location and identification of health professionals' practice sites is the first step in enhancing the availability of quality medical care to all regions of the state. Identification of specific areas of need is difficult, as provider data is often collected and housed separately through multiple organizations. This study provides a comparative review of the distribution of physicians, dentists, nurses, and psychologists and examines future provider needs of selected rural counties and regions within the state. METHODS: Data were compiled from multiple state and federal agencies. RESULTS: Twenty percent of Mississippi physicians are at or near retirement age. Several Mississippi counties could lose most or all of their physicians in the next few years. The distribution of health care workers is uneven across the state. CONCLUSION: Maldistribution of health care workers and the aging health care workforce impacts access to care, particularly in rural Mississippi counties. PMID- 12630640 TI - Increased incidence of the outbreak strain of Mycobacterium tuberculosis in the surrounding community after an outbreak in a jail. AB - BACKGROUND: Between 1995 and 1997, a tuberculosis outbreak occurred in a large, urban jail. We investigated whether the outbreak strain of Mycobacterium tuberculosis (M. tuberculosis) was circulating in the surrounding community after that outbreak. METHODS: We performed a retrospective cohort study of people with tuberculosis in Shelby County, TN, from January 1998 through August 1999, with molecular fingerprinting of M. tuberculosis strains. RESULTS: From January 1998 through August 1999, 23% of cases in the community involved a strain of M. tuberculosis that was indistinguishable from the previous jail outbreak strain. Twelve people (63%) with that strain had no history of recent incarceration. CONCLUSION: Two years after a tuberculosis outbreak in the jail, the outbreak strain was more prevalent in the surrounding community than it was during the jail outbreak. Jails can be important reservoirs of tuberculosis, which may subsequently circulate outside the institution. If efforts to eliminate tuberculosis are to be successful, the disease must be controlled successfully in such high-risk populations. PMID- 12630641 TI - Thoracic esophageal perforations. AB - BACKGROUND: Recognition of the importance of early diagnosis and aggressive, definitive surgical intervention has brought about a dramatic decline in mortality related to distal esophageal perforation. METHODS: We retrospectively analyzed all cases of thoracic esophageal perforation diagnosed at our hospital from September 1, 1979, through April 1, 2001. The study group consisted of 62 patients (43 men) with a mean age of 58.8 years (range, 20-92 yr). RESULTS: In the group of 39 patients with early diagnosis (< or = 24 h), hospital survival was 87%, which increased to 93% when early diagnosis was combined with aggressive surgical treatment Among the 23 patients with late diagnosis (> 24 h), survival approached 70%. Yet, in patients who were treated aggressively with surgery, survival was almost 90% despite delayed diagnosis. CONCLUSION: We recommend aggressive, definitive surgery for thoracic esophageal perforations, whether diagnosed early or late. A variety of options are discussed with regard to complicated presentations. PMID- 12630642 TI - Cutaneous metastasis: a meta-analysis of data. AB - BACKGROUND: Previous studies of the incidence and the most common causes of cutaneous metastasis have not led to a consensus. We compiled data from many retrospective studies and from patient data registries and autopsies to increase the total number of cases available for a larger analysis of this subject. This study was conducted to gain a better understanding of the true incidence of cutaneous metastasis, the tumors most commonly involved in this presentation, and the locations of such lesions. METHODS: A meta-analysis of cutaneous metastases from patient tumor registries and autopsic studies was performed. RESULTS: The overall incidence of cutaneous metastasis is 5.3%. The most common tumor to metastasize to the skin is breast cancer. The chest is the most common site of cutaneous metastasis. Every practitioner should be highly suspicious of acute onset, persistent, firm papulonodules, especially when they develop on the chest. CONCLUSION: This meta-analysis greatly increases the total number of cases available for the analysis of cutaneous metastases and provides a better overall view of this topic than was previously possible. PMID- 12630643 TI - Evaluation of a comprehensive loving support program among state Women, Infants, and Children (WIC) program breast-feeding coordinators. AB - BACKGROUND: Mississippi was selected as a pilot state in the national breastfeeding promotion campaign titled Loving Support Makes Breastfeeding Work (LSMBW). To reinforce the national LSMBW project, the Mississippi Women, Infants and Children (WIC) Breastfeeding Promotion Project Team developed a comprehensive program that included patient and family education, staff training, public awareness activities, health professional outreach, and partnerships with the community. The program also implemented a breastfeeding-friendly clinic environment project and a videotape project. This study was conducted to evaluate the impact of Mississippi's LSMBW activities among WIC breastfeeding coordinators in the United States. METHOD: The cross-sectional study was performed with the use of a mailed, self-administered questionnaire. RESULTS: Representatives of 50 state WIC agencies returned the survey. Of these 50 agencies, 36 (72%) had effectively used education materials created by the state of Mississippi. Breastfeeding coordinators reported that among the campaign activities, staff training, community outreach, and peer counseling were most beneficial. They also identified the videotape project developed by Mississippi as useful in addressing barriers to breastfeeding and in training support groups, staff, and health care professionals. CONCLUSION: This study showed that Mississippi's outreach activities and motivational videotape had a positive impact on coordinators' promotion of breastfeeding. PMID- 12630645 TI - Lipodystrophy, insulin resistance, diabetes mellitus, dyslipidemia, and cardiovascular disease in human immunodeficiency virus infection. AB - The introduction of highly active antiretroviral therapy has significantly reduced morbidity and mortality in patients infected with the human immunodeficiency virus. Treatment with antiretroviral agents--protease inhibitors in particular--has uncovered a syndrome of abnormal fat redistribution, dyslipidemia, and impaired glucose metabolism, collectively termed lipodystrophy syndrome. The cause of the syndrome seems to be multifactorial; however, its exact mechanisms have yet to be elucidated. Accelerated risk for cardiovascular disease is likely to be a major concern in these patients in the future. At this time, no clinical guidelines are available for the prevention and/or the treatment of lipodystrophy syndrome. The available treatment options range from switching the different antiretroviral drugs and lifestyle modifications to the use of pharmacologic agents to treat patients with dyslipidemia, impaired glucose tolerance and/or diabetes, and changes in body composition. This review emphasizes the clinical features, potential molecular mechanisms, and treatment options for patients infected with human immunodeficiency virus who have lipodystrophy syndrome. PMID- 12630646 TI - Spinal metastasis as the initial manifestation of a nonsecretory glucagonoma. AB - Glucagonomas are rare functional endocrine tumors of the pancreas that classically present with symptoms of glucagon excess, including rash, hyperglycemia, diarrhea, and weight loss. Metastatic disease at presentation is common but is often limited to the liver and regional lymph nodes. We describe an unusual case of a patient with glucagonoma who presented with a pathologic vertebral fracture. This tumor had no evidence of active hormone secretion but tested positive for glucagon by immunohistochemical staining. PMID- 12630644 TI - Ambulatory care-sensitive conditions: clinical outcomes and impact on intensive care unit resource use. AB - BACKGROUND: We identified risk factors and clinical outcomes associated with ambulatory care-sensitive conditions requiring intensive care unit (ICU) admission. METHODS: This prospective cohort study included 4,144 patients admitted to the medical ICU of an urban teaching hospital during a 3-year period. RESULTS: A total of 627 patients were classified as having ambulatory care sensitive conditions (ie, potentially preventable ICU admissions). Black race, decreasing Acute Physiology and Chronic Health Evaluation II (APACHE II) score, younger age, female sex, and absence of immunodeficiency were independently associated with ambulatory care-sensitive conditions. Patients classified as having ambulatory care-sensitive conditions accounted for 2,006 ventilator days, 2,508 ICU days, and 5,392 hospital days. The hospital mortality rate was statistically lower for patients with ambulatory care-sensitive conditions than for patients without these conditions. Patients classified as having ambulatory care-sensitive conditions were also statistically more likely than other patients to lack health insurance and to sign out of the hospital against medical advice. CONCLUSION: Patients with ambulatory care-sensitive conditions account for a substantial portion of all admissions to the intensive care unit. These data suggest that interventions aimed at preventing such admissions could improve ICU bed use. PMID- 12630647 TI - Panhemispheric infarction: a complication of cuffed catheter. AB - The need for reliable vascular access remains the Achilles heel of hemodialysis. Complications of vascular access are a leading cause of morbidity and mortality in patients who undergo hemodialysis, especially in those patients with end-stage renal disease. Among methods of vascular access, arteriovenous fistulae have the lowest rate of infection and should be the access of choice when vascular anatomy permits. Also, the incidence of staphylococcal infections in patients infected with human immunodeficiency virus is increasing. To emphasize the need to use arteriovenous fistula access for hemodialysis whenever possible, we report the case of a patient with end-stage renal disease and human immunodeficiency virus infection who died as a result of panhemispheric infarction and uncal herniation as a result of fulminant staphylococcal bacteremia caused by central venous catheter sepsis. PMID- 12630648 TI - Membranous glomerulonephritis in a patient with choriocarcinoma: case report. AB - A 26-year-old woman presented at our hospital with pretibial edema, swelling, and vaginal bleeding. She had nephrotic-level proteinuria and was diagnosed with choriocarcinoma after a histologic examination of her uterine curetting was performed. The renal biopsy specimen was compatible with membranous glomerulonephritis. The renal pathology in this case was thought to be secondary to choriocarcinoma because the nephrotic syndrome regressed after hysterectomy. We briefly present this first case of choriocarcinoma causing membranous glomerulonephritis. PMID- 12630649 TI - Phenytoin-induced toxic cholestatic hepatitis in a patient with skin lesions: case report. AB - Phenytoin is a highly effective and widely prescribed anticonvulsant agent, but it can be associated with dose-related side effects and hypersensitivity reactions. We present a case of phenytoin-induced cholestatic hepatotoxicity in a 47-year-old woman who had exfoliative dermatitis, an increase in liver enzymes with a cholestatic pattern, and eosinophilia after 25 days of phenytoin therapy. The diagnostic workup showed no other possible causes, and the results of a percutaneous liver biopsy were consistent with drug-induced toxic hepatitis. Within 3 weeks after discontinuing phenytoin therapy, her liver function tests returned to normal values. PMID- 12630650 TI - Pseudo-central hypothyroidism. AB - Central hypothyroidism is an exceedingly rare form of hypothyroidism that results from a variety of conditions affecting the hypothalamus and the pituitary gland. The classic biochemical abnormality seen in these patients includes a low serum level of circulating thyroxine (T4) concomitant with an inappropriately low level of thyrotropin. Because patients with isolated triiodothyronine (T3) toxicosis also present with this biochemical pattern, it is important to measure T3 levels in such patients before making a diagnosis of central hypothyroidism. PMID- 12630651 TI - Staphylococcal acalculous cholecystitis in a child. AB - Acute acalculous cholecystitis is inflammation of the gallbladder in the absence of gallstones. It usually occurs in critically ill patients and is rare in the pediatric age group. We describe a 12-year-old boy who presented with fever, jaundice, and abdominal pain and was found to have acute acalculous cholecystitis, sacroiliitis, and pelvic osteomyelitis associated with bacteremia as a result of Staphylococcus aureus. Antibiotic therapy without surgical intervention was effective. A high index of suspicion is required to make an early diagnosis and institute appropriate treatment for children with this condition. Although cholecystectomy has been considered the standard therapy, medical treatment alone can be successful. PMID- 12630652 TI - Prolactinoma and other head and neck tumors after scalp irradiation. AB - Tumors of the thyroid and parathyroid glands may develop together or separately in patients who previously have been exposed to head and neck irradiation. Whether cranial irradiation confers an increased risk for pituitary adenoma remains unknown. We report the case of a 52-year-old woman who was treated during childhood for tinea capitis with scalp irradiation and later in life developed a prolactin-secreting tumor, a parathyroid adenoma, a benign thyroid lesion, and a basal cell carcinoma of the skin. She was treated successfully with bromocriptine and surgical removal of the parathyroid adenoma. Molecular analysis of the parathyroid tissue failed to demonstrate any abnormality of the multiple endocrine neoplasia Type 1 gene. This case report is the first to describe a prolactin-secreting tumor that developed in association with other endocrine neoplasia after head and neck irradiation. Our case suggests that multiple endocrine neoplasia may develop in a sporadic pattern after scalp irradiation. PMID- 12630653 TI - First-degree atrioventricular block and restrictive physiology as cardiac manifestations of Fabry's disease. AB - Fabry's disease is an X-linked disorder of glycosphingolipid catabolism related to the defective activity of glycosphingolipid, mainly ceramide trihexoside, in the vascular smooth muscle, myocardium, cells of the sympathetic central nervous system, and epithelial cells of renal glomeruli. We describe a young man who had Fabry's disease and unusual electrocardiographic and echocardiographic patterns at admission for treatment of left leg cellulitis. Findings included a prolonged PR interval and a right bundle branch block pattern, no echocardiographic signs of septal or hypertrophic cardiomyopathy, and a restrictive physiologic pattern. This pattern of electrocardiographic and echocardiographic characteristics of Fabry's disease has not been reported previously and should be added to the other cardiac manifestations of Fabry's disease. PMID- 12630654 TI - Central venous catheter infection in a child: case report and review of Kluyvera infection in children. AB - Kluyvera is an opportunistic pathogen that can occur in immunosuppressed as well as immunocompetent hosts. We report a case of Kluyvera species infection involving a central venous catheter, and we review the literature on Kluyvera infections in children. Our case demonstrates that removal of the central venous catheter was necessary to eradicate the infection and hasten the resolution of refractory neutropenia. The spectrum of disease due to Kluyvera infection in children includes central venous catheter infection and/or sepsis, urinary tract infection, enteritis, and, in one instance, fatal peritonitis. It is clear on the basis of our case report that uncommon, opportunistic organisms such as Kluyvera can be significant pathogens. PMID- 12630655 TI - Neuropsychological characteristics of patients in a hospital-based eating disorder program. AB - The existence of cognitive deficits associated with eating disorders has been debated for some time. The present study investigated cognitive impairments in a large sample of patients with anorexia nervosa from an inpatient treatment program. Fifty-nine women with anorexia nervosa were given a battery of neuropsychological tests assessing multiple cognitive domains. Over half of the patients had mild cognitive impairments in two or more neuropsychological tasks and approximately one-third failed two or more tasks. Depression level and body mass were not associated with cognitive impairment. Whether effective restoration of weight and resolution of core psychopathology contribute to reversal of cognitive deficits requires further research. PMID- 12630656 TI - The influence of cognitive impairment and psychiatric symptoms on daily functioning in nursing facilities: a longitudinal study. AB - The influence of psychiatric symptoms and cognitive status on daily living skills was explored among persons with dementia residing in skilled nursing facilities. Psychiatric symptoms were examined in three domains: 1) depression, 2) psychosis, and 3) behavioral disturbances. Twenty-eight subjects were followed for a 1-year duration in rural nursing facilities. The relationship between psychiatric symptoms, cognition, and daily living skills was assessed using correlation and regression analyses. Over a one-year period, cognitive and functional impairment progressed significantly. In contrast, psychiatric symptoms across all three domains did not necessarily persist or increase. Deterioration in cognitive function was significantly correlated with reduced functional status both cross sectionally and longitudinally. Among psychiatric symptoms, only behavioral disturbances were significantly associated with functional impairment cross sectionally. Cognitive decline and loss of social functioning may constitute the greatest source of disability among persons in nursing facilities. Furthermore, both impairments increase over time in contrast to psychiatric symptoms, which appear to have a more variable course. These findings may have implications for treatment interventions that seek to enhance functioning in the skilled nursing setting. PMID- 12630657 TI - Three- to four-year follow-up to an open trial of nefazodone for combat-related posttraumatic stress disorder. AB - Multiyear (37-51 months) follow-up data was obtained on patients who had participated in an open label trial of nefazodone that originally showed nefazodone may be useful for symptom management in posttraumatic stress disorder (PTSD) patients. Ten patients with combat-related DSM-IV posttraumatic stress disorder (PTSD) entered an open-label 12-week trial of nefazodone, beginning with 100 mg/day and increasing as necessary to achieve a maximal response or until reaching a maximum dosage of 600 mg/day. All 10 patients were followed for over 3 4 years and used nefazodone with dosages of 400-600 mg a day. The entire dosagewas shifted to bedtime to facilitate sleep in 7 patients. Data on PTSD symptoms, depression, sleep, and anger were examined. Nefazodone was well tolerated and no significant changes in sexual function were reported. All participants reported compliance with the prescribed nefazodone over 3-4 years. Nine patients reported that it remained effective, and expressed a desire to remain on the medication. On the basis of clinician global impression ratings (compared to baseline), 10 patients were rated as much improved at 12 weeks. Seven of the 10 patients continued to be much improved, 2 were minimally improved, and 1 was rated as worse (compared to baseline assessment) on 3-4-year follow-up. At 3-4-year follow-up, improvements in PTSD symptoms, sleep, and anger were maintained. These improvements were statistically significant with moderate to-large effect sizes. These data suggest that clinical improvement in PTSD patients administered nefazodone may be maintained with continued treatment. The medication was tolerated well in long-term treatment, compliance was high, and improvement was maintained over several years. Length of treatment, appropriate dose, long-term efficacy, and compliance are all clinically significant issues with little guiding data available. Controlled studies are needed to (a) further investigate the long-term efficacy of nefazodone in the treatment of PTSD; (b) provide information for length of treatment guidelines; and (c) document if discontinuation is possible and efficacious. PMID- 12630658 TI - Combining lithium and anticonvulsants in bipolar disorder: a review. AB - While the benefits of lithium in bipolar disorder are evident, its limitations as monotherapy are well recognized, particularly in bipolar depression. This has propelled trials of combined lithium-anticonvulsant therapy in many bipolar patients. The present review of the English-language literature examines both controlled and open studies of such combination therapy, including the risk of drug-drug interactions. Trials of lithium plus either carbamazepine or divalproex have generally produced favorable results, although increased rates of adverse effects may reduce treatment adherence. More recent reports suggest that lithium may be safely and effectively combined with lamotrigine, and perhaps with topiramate, although controlled studies are required. The combined use of lithium with newer, putative mood stabilizers, such as zonisamide or levetiracetam, cannot yet be recommended, but is an important area for future research. Provisional recommendations for combined treatment are provided. PMID- 12630659 TI - New anticonvulsants: a review of applications for the management of substance abuse disorders. AB - Substance abuse and related disorders are a major problem in the United States and elsewhere. In many cases, pharmacotherapy has been shown to be useful in the treatment of substance abuse and related disorders, both in detoxification and in the prevention of relapse. However, currently used medications are not ideal, due either to their effectiveness being less than desirable or their side effect profiles being greater than desirable, or both. Medications with increased effectiveness or decreased side effects are needed. This article reviews three anticonvulsant medications, gabapentin, oxcarbazepine, and tiagabine, for possible utility in the treatment of substance abuse and related disorders. The following aspects of each medication are discussed: metabolism and kinetics, dosage and administration, possible interactions, side effects, and applications to substance abuse treatment; the last subject includes discussion of management of withdrawal symptoms, treatment of dual diagnosis, management of substance abuse sleep disorders, direct suppression of substance abuse/relapse prevention, and abuse potential. Each medication appears to have potential advantages for these uses, and further study of these medications in treatment of substance abuse disorders is recommended. PMID- 12630660 TI - Central pontine myelinolysis manifested by temporary blindness: a possible complication of lithium toxicity. AB - Central pontine myelinolysis (CPM) is removal of myelin material from neural elements in a way that is not clearly known as yet. In this case of CPM, blindness was encountered and was thought to be "hysterical." The blindness went away after four months. After reviewing the literature we suggest the CPM was a complication of lithium toxicity which affected the lateral geniculate nucleus which produced blindness. PMID- 12630661 TI - Risk from lithium with acetylcholinesterase inhibitor. PMID- 12630662 TI - What we currently know about the structure and function of the p53 homologue - p73 protein: facts, hypotheses and expectations. PMID- 12630663 TI - Adult stem cells and their importance in cell therapy. AB - For their unique properties stem cells promise to be of universal use in clinical medicine, especially in regeneration of many organs and tissues in the human body. This attractive subject receives an ever growing attention of specialists from different branches of science and, no doubt, will present one of the most studied trends in medicine in the new millennium. In this communication, the authors discuss two main sources of human stem cells potentially suitable for cell-based therapy. The first are the cells obtained from embryonic tissues- embryonic stem cells, the second are the cells derived from adult tissues--adult stem cells. Presently, harvesting and therapeutic use of embryonic stem cells are associated with many problems both methodical and ethical. Utilization of adult stem cells in cell-based therapy is a certain solution in the current state of replacement therapy. Still, we have to be aware that this is not a compromise but one of the most prospective ways to treat a variety of serious diseases. To date, it is not yet clear which way would be more suitable and it is up to us which way we choose for the benefit of millions of patients. Considering the current state of knowledge, it is impossible yet to predict which stem cells--embryonic or adult--or therapeutic approaches would yield the best results. Much research is to be done and verified in practice and, at the same time, ethical problems must be resolved. PMID- 12630664 TI - Proteomic analysis of radiation-induced alterations in L929 cells. AB - In this study we examined the protein expression profiles in X-irradiated L929 cells to get insight into how mammalian cells respond to radiation-induced cell damage. L929 cells were irradiated with the dose of 6 Gy and cell lysates were collected at different time intervals (20 min, 12, 24, 36, 48 and 72 h). The extracted proteins were separated by 2-DE and quantified using computerized image analysis. Proteins exhibiting significant abundance alterations when comparing irradiated to unirradiated cells were identified by mass spectrometry. Using the proteomics approach we detected 47 proteins that exhibited a significant radiation-induced increase or decrease in the course of 72 h. From this group of spots 28 proteins were identified by mass spectrometry and of these 24 proteins exhibited minimally 2-fold differences in mean abundance values in comparison to controls. The identified proteins represent diverse sets of proteins participating either in protective and reparative cell responses or in induction of apoptosis and oncogenesis. The results document that proteomics is a useful method for unravelling the molecular mechanisms involved in cell reaction to ionizing radiation. PMID- 12630666 TI - Two dynamic morphotypes of sarcoma cells, asymmetric stellate and triangle with leading lamella, are related to malignancy. AB - A notion of the dynamic morphotype was developed as a conjunction between cell shape and migration. This enabled the investigation of the relationship between malignancy and patterns of dynamic morphology in neoplastic cells in vitro. Time lapse cinemicroscopy was used to analyse the cell behaviour of three rat neoplastic cell lines (K2, T15, and A8), differing in metastatic potential, that were instrumental in revealing a coincidence between high migratory activity and appearance of the 3D structure of actin cables in high-malignant A8 cells (Pokorna et al., 1994). A set of criteria was established for visual classification of cell morphology. Matching the pattern of cell morphology with locomotory activity led to identification of four dynamic morphotypes. Cell speed was determined by tracking and the dynamic morphotypes assigned by the operator. All the three cell populations were studied for incidence of the dynamic morphotypes in culture media differing in pH: 6.6 simulating acid extracellular condition in tumours, physiological 7.4, and alkaline 8.2. The results showed that acid pH stimulated motile activity in the intermediate-malignant T15 and most malignant A8 cells. The T15 and A8 cells also manifested a prolonged continuation of fast locomotion in the early G1 phase and displayed a prevalence of two fast moving dynamic morphotypes: asymmetric stellate and triangle with leading lamella. PMID- 12630665 TI - Local IFN-gamma therapy of HPV16-associated tumours. AB - We have examined whether peritumoral administration of IFN-gamma can inhibit growth of HPV16-associated, MHC class I- tumour MK16/1/IIIABC (MK16) transplanted in syngeneic mice. It has been found that peritumoral administration of recombinant IFN-gamma performed on days 0-11 after tumour challenge inhibited growth of MK16 s.c. tumour transplants. If the therapy with IFN-gamma was started when the tumours had already reached a palpable size, the IFN-gamma administration was without any effect. To investigate the antitumour effects of IFN-gamma in a clinically more relevant setting, surgical minimal residual tumour disease was utilized. Subcutaneously growing MK16 carcinomas, 8-12 mm in diameter, were removed and the operated mice were injected with IFN-gamma on days 3-14 after the operation at the site of surgery. Treatment with IFN-gamma resulted in a moderate, reproducible, but statistically insignificant inhibition of tumour recurrences. In the next experiments we have addressed the question whether the tumour-inhibitory effect of IFN-gamma was due to the upregulation of MHC class I molecule expression on MK16 tumour cells. IFN-gamma-treated and control mice were sacrificed, their tumours were explanted, and the expression of MHC class I molecules on the MK16 tumour cells was examined. As presumed, the MHC class I expression on the cells of IFN-gamma-treated tumours, as well as on their lung metastases, was upregulated. However, an unexpected moderate upregulation of the MHC class I expression was also observed on MK16 tumours from the control, exogenous IFN-gamma-uninjected mice. Cytofluorometric analysis of the in vivo transplanted MK16 tumours from both groups has excluded that the increased percentage of the MHC class I molecules on the tumour cell populations could be due to the infiltration of the tumours with MHC class I+ leukocytes, since no expression of MHC class II, CD11b, CD80/CD86, and CD11c molecules in the MK16 cell population was observed. PMID- 12630667 TI - Interaction of Borrelia burgdorferi sensu lato with Epstein-Barr virus in lymphoblastoid cells. AB - Since the possibility of interruption of latent EBV infection has been suggested by the induction of the lytic virus cycle with chemical substances, other viruses, and by immunosuppression, we hypothesized that the same effect might happen in B. burgdorferi sensu lato infection as happens in Lyme disease patients with positive serology for both agents. We have observed EBV replication in lymphoblastoid cells after superinfection with B. garinii and B. afzelii strains after 1 and 4 h of their interaction. We found that viral and borrelial antigens persisted in the lymphoblasts for 3 and 4 days. Morphological and functional transformation of both agents facilitate their transfer to daughter cells. Association with lymphoblasts and internalization of B. garinii by tube phagocytosis increased replication of viruses more successfully than B. afzelii and chemical inductors. Demonstration of such findings must be interpreted cautiously, but may prove a mixed borrelial and viral cause of severe neurological disease. PMID- 12630668 TI - Improving patient satisfaction and acne severity in patients with mild to moderate acne: the BEST study. PMID- 12630669 TI - Study design and selection criteria in the BEST study. AB - Patients with acne should receive prompt and effective treatment that satisfies their needs. The combination of benzoyl peroxide/clindamycin has demonstrated efficacy for the treatment of patients with acne vulgaris. The BenzaClin (benzoyl peroxide/clindamycin topical gel) Efficacy and Satisfaction Trial (BEST), a large, open-label, multicenter study, evaluated patient satisfaction in response to 8 weeks of treatment with benzoyl peroxide/clindamycin topical gel in patients with mild to moderate acne who were dissatisfied with their current acne treatment regimen. Patients eligible for the study were at least 12 years of age and had rated satisfaction with their prior acne therapy as low (0-4) on an 11 point visual analog scale (0 = not satisfied to 10 = very satisfied). Patients with clinically significant cardiovascular, pulmonary, renal, or endocrine disease were excluded from this study, as were those with a hypersensitivity to study drug components. Study variables evaluated at baseline and after 8 weeks of treatment with benzoyl peroxide/clindamycin topical gel included patient satisfaction; acne severity, evaluated with the Global Acne Grading System (GAGS); evaluation of the social aspects of living with acne, measured with the Acne Quality of Life (AQOL) scale; and Physician Global Assessment (PGA) of patient response to treatment, measured on a 5-point scale (0 = worse to 4 = marked improvement). This open-label study design allowed for assessment of patient satisfaction in response to treatment in a large patient population (n = 1,389) in a real-world setting. PMID- 12630670 TI - Overall results of the BEST study following treatment of patients with mild to moderate acne. AB - In patients with acne, assurance of satisfaction with therapy is a significant factor in helping patients cope with their condition. Therefore, the BenzaClin (benzoyl peroxide/clindamycin topical gel) Efficacy and Satisfaction Trial (BEST) was developed to evaluate patient satisfaction following 8 weeks of treatment with benzoyl peroxide/clindamycin topical gel in patients with mild to moderate acne who were dissatisfied with their current acne therapy. Additional evaluations included efficacy, evaluated with the Global Acne Grading System (GAGS), and the impact of therapy on the social aspects of living with acne, measured with the Acne Quality of Life (AQOL) scale. Results demonstrated significant improvements for all study variables. Patient satisfaction, which was rated from 0 (not satisfied) to 10 (very satisfied) increased almost 3-fold from baseline to week 8 (P < or = .0001). The mean overall AQOL score also improved significantly by a mean of 2.2 points (P < .0001). Acne severity was reduced to almost half of its baseline value (measured using the GAGS score), indicating a significant treatment effect of benzoyl peroxide/clindamycin topical gel (P < or = .0001). Furthermore, as indicated by the Physician Global Assessment (PGA) performed at week 8, acne improved in 94% of patients. Improved patient satisfaction was most likely related to the significant improvements in both efficacy and quality of life (QOL). Overall results of the study suggest that benzoyl peroxide/ clindamycin topical gel is effective for the treatment of acne in patients who are dissatisfied with their current therapy. PMID- 12630671 TI - The BEST study: evaluating efficacy by selected demographic subsets. AB - Acne therapy often requires continuous treatment with a combination of agents. Patient satisfaction with treatment ensures proper compliance, which ultimately leads to a successful outcome. The BenzaClin (benzoyl peroxide/clindamycin topical gel) Efficacy and Satisfaction Trial (BEST) was conducted to determine changes in the degree of satisfaction after using benzoyl peroxide/clindamycin topical gel in patients with mild to moderate acne who were not satisfied with their current therapy. Reported in this subanalysis are results stratified according to selected patient demographic subsets (18 years and younger, 21-30 years; male, female; white, black, Hispanic, and Asian; and patients with mild acne). Efficacy variables included patient satisfaction, acne condition and severity, quality of life (QOL), and physician assessment of treatment response. Significant increases in patient satisfaction and improvement in acne condition were demonstrated with the use of benzoyl peroxide/clindamycin topical gel by male and female patients of the ages and races evaluated, as well as by patients with mild acne (P < or = .0001 for all patient subsets). Patients across genders and the specified age groups and races, as well as patients with mild acne, reported significant (P < .05) improvements in QOL following treatment with benzoyl peroxide/clindamycin topical gel. The majority of patients in most subsets, including those with mild acne, demonstrated marked improvements as measured by Physician Global Assessment (PGA) in response to treatment. In summary, patients who have been dissatisfied with their treatment regimen may be more satisfied with the use of this benzoyl peroxide/clindamycin topical gel regardless of age, gender, or race. PMID- 12630673 TI - Implications of the BEST study. PMID- 12630672 TI - The BEST study: results according to prior treatment. AB - Combination topical products may provide the best available treatment option to patients with acne because the majority of these products not only combine drugs with 2 distinct mechanisms of action but also avoid systemic adverse events that may be associated with oral therapies. Recently, the BenzaClin (benzoyl peroxide/ clindamycin topical gel) Efficacy and Satisfaction Trial (BEST) was conducted. This large, open-label, multicenter study assessed patient satisfaction, treatment efficacy, and quality of life (QOL) in response to 8 weeks of treatment with benzoyl peroxide/clindamycin topical gel. Patients who were previously dissatisfied with their current acne therapy were enrolled in the study. This paper presents results stratified by prior treatment in patients who were receiving benzoyl peroxide, clindamycin, other topical antibiotics, or retinoids prior to study enrollment. Mean patient satisfaction scores were significantly increased regardless of which prior treatments were used (P < or = .0001 for all groups). Furthermore, acne severity was significantly improved among all groups (P < or = .0001), with more than 90% of most groups experiencing marked, moderate, or mild improvements in their acne by the end of the study. Patient satisfaction is an important element in the experience of acne because it is usually linked to improved efficacy; therefore, assurance of patient satisfaction with therapy is a significant consideration in the treatment of this illness. PMID- 12630674 TI - Bicyclic nucleoside inhibitors of varicella-zoster virus: effect of terminal aryl substitution in the side-chain. AB - We have previously reported the discovery and preliminary structure-activity relationships of a new class of inhibitors of varicella-zoster virus (VZV). These novel furanopyrimidine nucleosides bear unusual bicyclic base moieties and exhibit complete specificity for VZV. Limited in vitro cytotoxicity has been detected and the bicyclic nucleosides are now well established as a new family of potent antivirals. Our initial studies revealed an absolute requirement of a long alkyl side-chain, with an optimal chain length of C8-C10, for antiviral activity. Following further studies, we recently reported a significant enhancement of both antiviral potency and selectivity by the inclusion of a phenyl group within the alkyl side-chain of these compounds. The new lead p-alkylphenyl analogues displayed EC50 values below 1 nM versus VZV and selectivity index values >1000000. We herein report the synthesis and characterization of a further series of alkylaryl analogues bearing terminal phenyl groups with varying n-alkyl side chain lengths. Synthesis of the target bicyclic systems involved the Pd-catalysed coupling of terminal acetylenes with 5-iodo-2'-deoxyuridine to give intermediate 5-alkynyl nucleosides which were then cyclized in the presence of copper (I) iodide. The current compounds display excellent selectivity for VZV with no detectable in vitro cytotoxicity but despite being chemically isomeric with the previous lead p-alkylphenyl analogues, the compounds reported herein exhibit only moderate antiviral activities. A possible correlation between antiviral activity and conformational freedom of the side-chain is discussed. PMID- 12630675 TI - Inhibition of HIV-1 replication by an HIV-1 dependent ribozyme expression vector with the Cre/loxP (ON/OFF) system. AB - Antiviral strategies to inhibit HIV-1 replication have included the generation of gene products that provide the intracellular inhibition of an essential viral protein or RNA. When used in conjunction with the HIV-1 long terminal repeat (LTR), an inducible promoter dependent on the virus-encoded trans-activator (tat), relatively high background activity is still observed in the absence of tat (Caruso & Klatzmann, 1992; Dinges et al., 1995). In order to circumvent this problem, we used the Cre/loxP (ON/OFF) recombination system as a tool for our investigation. In the present study, we constructed a loxP-cassette vector with the ribozyme (Rz) expression portion under the control of the tRNAi(Met) promoter between two loxP sequences (plox-Rz-U5). We also constructed an HIV-1 LTR promoter-driven Cre recombinase gene (pLTR-Cre). These vectors were triple transfected into HeLa CD4 cells with the HIV-1 pseudotype viral expression vector. Basal activity was not detectable before HIV-1 infection. The LTR dependent Cre protein product in HIV-1 infected HeLa CD4 cells expressed the ribozyme by inducing loxP homologous recombination, which strongly inhibited the HIV-1 gene expression. These results demonstrate the potential of an anti ribozyme with the Cre/loxP system for controlling HIV-1 infection via gene therapy. PMID- 12630677 TI - Synthesis and anti-HIV activity of carbamates of antiviral agent stavudine. AB - An efficient synthesis of carbamate analogues of the NRTI compound stavudine, has been achieved in five steps starting from commercially available thymidine. The synthesis involves conversion of thymidine into stavudine followed by condensation with carbaimidazole derivative obtained from various aromatic and heterocyclic amines in dimethylformamide solvent. The analogues thus obtained were further purified by crystallization to furnish analytically pure products. Examination of biological activity of these carbamate derivatives of stavudine showed that they inhibited HIV replication only at micro-molar concentrations. PMID- 12630676 TI - A novel substance purified from Perilla frutescens Britton inhibits an early stage of HIV-1 replication without blocking viral adsorption. AB - Pf-gp6, a 6 kDa anti-degranulation glycoprotein purified from the extract of Perilla frutescens, was examined for its antiviral activity against HIV-1 and HIV 2 in vitro. HIV-1-induced cytopathic effect and proviral DNA synthesis were inhibited in the presence of Pf-gp6. The 50% inhibitory concentrations of Pf-gp6 for various HIV-1 strains, including clinical isolates and CCR5-using (R5) HIV-1, ranged between 1.3 and 71.0 microg/ml, depending on the combination of viral strain and host cell. Furthermore, Pf-gp6 did not directly inactivate infectious viral particles. A time-of-addition experiment revealed that Pf-gp6 lost its activity before zidovudine but after the CXCR-4 antagonist AMD3100 during the early stage of viral infection. Although the pinpoint target of Pf-gp6 remains to be elucidated, it may interfere with a step between viral entry and reverse transcription. PMID- 12630678 TI - Imino sugars that are less toxic but more potent as antivirals, in vitro, compared with N-n-nonyl DNJ. AB - Imino sugar glucosidase inhibitors have selective antiviral activity against certain enveloped, mammalian viruses. Deoxynojirimycins (DNJs) modified by N alkylation to contain a nine carbon atom side chain (N-n-nonyl-deoxynojirimycin; N-nonyl-DNJ, NN-DNJ) were shown to be, for example, at least 20 times more potent in inhibiting hepatitis B virus (HBV) and bovine viral diarrhoea virus (BVDV) in cell based assays than the non-alkylated DNJ. These data suggested that modification of the alkyl side chain could influence antiviral activity. Previous work has focused on varying side chain length. In this report, the influence of side chain branching and cyclization upon toxicity and antiviral activity was explored. Briefly, using a virus secretion assay for HBV and a single step growth (yield reduction) assay for BVDV, 14 different DNJ-based sugars, possessing various N-alkyl substitutions, were tested for antiviral activity. Of the series, N-methoxy-nonyl-DNJ and N-butyl-cyclohexyl DNJ were determined to have the best selectivity index against BVDV and HBV, with the N-methoxy analogue being the most potent with micromolar antiviral activity. The results of this antiviral survey and the implications for the mechanism of action and ultimate therapeutic potential of the DNJ-based imino sugars is provided and discussed. PMID- 12630680 TI - Selection of a thiazole urea-resistant variant of bovine viral diarrhoea virus that maps to the RNA-dependent RNA polymerase. AB - By passing wild type bovine viral diarrhoea virus (BVDV) in increasing concentrations of DPC-A69280-29, a thiazole urea class compound that inhibits BVDV replication, we were able to select several variants of BVDV that exhibited decreased susceptibility to this compound. When the non-structural genes of these variants were sequenced and compared with wild type, only one change was common to all the variants that also exhibited resistance to DPC-A69280-29 (>10-fold increase in IC50). This change was a T-to-A transversion at position 11198 of the BVDV genome, which would cause a predicted substitution of isoleucine for phenylalanine at amino acid 78 of the RNA-dependent RNA polymerase (RdRp). This substitution would occur in a region of the BVDV RdRp which has been proposed to be important for the formation of the RdRp homodimer that is essential for the activity of the enzyme. However, since DPC-69280-29 inhibits BVDV replication by interfering with the initiation of viral RNA synthesis, we discuss the possibility that this region of the BVDV RdRp also may play a role in the initiation process. Furthermore, since this region is located fairly close to the template RNA, we also propose that the role it plays may involve either template selection, stabilization or processivity. PMID- 12630681 TI - MR physics of body MR imaging. AB - This article reviews the major challenges of body imaging, describes the problems that arise from motion, and the many attempts at reducing this problem. Fast imaging sequences and approaches to reducing the data acquired without sacrificing image quality are described. PMID- 12630679 TI - Short-term treatment with novel ribonucleotide reductase inhibitors Trimidox and Didox reverses late-stage murine retrovirus-induced lymphoproliferative disease with less bone marrow toxicity than hydroxyurea. AB - We evaluated the ability of a short course of treatment with the ribonucleotide reductase (RR) inhibitor hydroxyurea (HU) and two novel RR inhibitors Trimidox (TX) and Didox (DX) to influence late-stage murine retrovirus-induced lymphoproliferative disease. LPBM5 murine leukaemia virus retrovirus-infected mice were treated daily with HU, TX or DX for 4 weeks, beginning 9 weeks post infection, after development of immunodeficiency and lymphoproliferative disease. Drug effects on disease progression were determined by evaluating spleen weight and histology. Effects on haematopoiesis were determined by measuring peripheral blood indices (white blood cells and haematocrit) and assay of femur cellularity and femoral and splenic content of colony-forming units granulocyte-macrophage (CFU-GM) and burst-forming units-erythroid (BFU-E). HU, TX and DX partially reversed late-stage retrovirus-induced disease, resulting in spleen weights significantly below pre-treatment values. Spleen histology was also improved by RR inhibitor treatment (DX>TX>HU). However, as expected, HU was significantly myelosuppressive, inducing a reduction in peripheral indices associated with depletion of femoral CFU-GM and BFU-E. In contrast, although TX and DX were moderately myelosuppressive, both drugs were significantly better tolerated than HU. In summary, short-term treatment in late-stage murine retroviral disease with HU, TX or DX induced dramatic reversal of disease pathophysiology. However, the novel RR inhibitors TX and DX had more effective activity and significantly less bone marrow toxicity than HU. PMID- 12630682 TI - Cardiac MR imaging. AB - There has been tremendous progress for MR imaging depiction of cardiac morphology and function. Further advances toward achieving faster acquisition with real-time imaging, higher resolution for plaque imaging, and quantitative analysis are taking place at a rapid pace. PMID- 12630683 TI - MR angiography of the chest. AB - Using the described strategies all relevant disease processes of the thoracic vessels can be fully depicted using contrast-enhanced three-dimensional MRA. The aorta and the major neck and arm vessels are well visualized. Vascular pathologies, such as aneurysms, dissections, and occlusions, are readily recognized. With the implementation of high-performance gradients, three dimensional MRA of the pulmonary vasculature has become possible even in dyspneic patients. Congenital lesions, such as coarctations, are particularly well suited for analysis with these techniques. PMID- 12630685 TI - MR imaging of liver tumors. AB - The article reviews the current MR imaging techniques commonly utilized for imaging liver tumors. Breath-hold T1-weighted GRE sequences, FSE T2-weighted sequences, and properly timed contrast-enhanced 3D SGE are important for lesion characterization. New liver-specific contrast agents improve lesion detection and are useful in lesion characterization. PMID- 12630684 TI - Breast MR imaging. AB - Contrast-enhanced breast MR imaging has made significant progress since its introduction into the radiological literature in 1989. The techniques and technology continue to be refined, and understanding of the interpretation strategies has improved dramatically. Clinical applications in difficult diagnostic cases and the evaluation of the extent of breast cancer are now being practiced in many centers worldwide. There is great excitement over the potential for breast MR imaging to address the problem of screening high-risk women. Despite all of the progress made over the past years, however, there is still a significant amount of work ahead before a clear understanding of how this technique will affect the health care of women is obtained. PMID- 12630686 TI - MR imaging of diffuse liver disease. AB - MR imaging is able to demonstrate and distinguish the full variety of benign and malignant diffuse liver diseases. PMID- 12630687 TI - MR cholangiopancreatography. AB - Although MRCP is still an evolving technique, it has established itself as clinically useful and comparable with ERCP for the evaluation of various biliary or pancreatic ductal diseases. MRCP is not only comparable with ERCP in its diagnostic ability, but it has the tremendous advantage of being noninvasive. Furthermore, MR imaging is useful in patients with incomplete or failed ERCP, and in patients with certain biliary or gastrointestinal surgical procedures it is the imaging modality of choice. ERCP will remain an extremely important modality because of the great clinical importance for interventional biliary procedures with this technique. Nonetheless, MRCP may in the near future replace most of the diagnostic imaging of the biliary tree, with diagnostic results even more improved with further developments of hardware and technique. PMID- 12630688 TI - MR cholangiopancreatography: evaluation of common pancreatic diseases. AB - In the evaluation of common pancreatic diseases, MRCP is a noninvasive alternative to ERCP. Ductal anatomy can be ascertained without risk of complications. MRCP is valuable in defining common anatomic variants, determining the state of the pancreatic duct in pancreatitis, and characterizing neoplasms, especially combined with other MR imaging sequences. With the advent of MRCP, techniques requiring endoscopy and percutaneous access are largely reserved for histologic diagnosis and treatment, or for cases in which MRCP fails to establish a diagnosis. PMID- 12630689 TI - MR angiography of the abdominal aorta and peripheral vessels. AB - Contrast-enhanced MRA can be an accurate and reliable method for the arterial evaluation of the abdominal aorta and peripheral vessels. This technique can be adapted for a variety of anatomic regions. The basic issues relate to proper synchronization of imaging with peak arterial enhancement and to optimization of voxel dimensions for adequate depiction of the arterial structures. PMID- 12630690 TI - MR imaging of the kidneys and adrenal glands. AB - By performing a comprehensive MR imaging examination, it is not only possible accurately to characterize and stage cystic and solid lesions of the kidney, but also to provide important preoperative information to the surgeon. In addition, MR imaging can characterize many adrenal lesions and frequently can obviate the need to obtain biopsies. The continued development and growth of MR technology combined with the current trend toward minimally invasive surgery will expand the role of MR imaging in the future. PMID- 12630692 TI - MR imaging of the female pelvis. AB - MR imaging has become a valuable modality in the evaluation of the female pelvis. In many cases, it follows the performance of HSG or US. These cases include infertility and pelvic pain. In some cases, it should serve as the test of choice. This is true in the local staging of cervical cancer and the evaluation of pain or disability in the pregnant patient. Finally, in the evaluation of advanced gynecologic cancers, it is usually a secondary choice with CT preferred. PMID- 12630691 TI - MR imaging of the bladder. AB - MR imaging of the bladder can now be routinely and easily performed with consistent quality. The latest techniques provide high-resolution images of the soft tissue contrast and the ability to perform dynamic contrast imaging and functional pelvic floor studies. MR imaging fits in as a complimentary tool to cystoscopy and to conventional pelvic floor dynamic studies and can provide a unique imaging perspective of the bladder. It is important, however, to remain cognizant of the limitations of its use in tumor detection and discrimination. PMID- 12630693 TI - Surface plasmon resonance spectroscopy in the study of membrane-mediated cell signalling. AB - Peptide-membrane interactions contribute to many important biological processes such as cellular signaling, protein trafficking and ion-channel formation. During receptor-mediated signalling, activated intracellular signalling molecules are often recruited into receptor-induced signaling complexes at the cytoplasmic surface of the cell membrane. Such recruitment can depend upon protein-protein and protein-lipid interactions as well as protein acylation. A wide variety of biophysical techniques have been combined with the use of model membrane systems to study these interactions and have provided important information on the relationship between the structure of these proteins involved in cell signalling and their biological function. More recently, surface plasmon resonance (SPR) spectroscopy has also been applied to the study of biomembrane-based systems using both planar mono- or bilayers or liposomes. This article provides an overview of these recent applications, which demonstrate the potential of SPR to enhance our molecular understanding of membrane-mediated cellular signalling. PMID- 12630694 TI - Synthetic peptides mimicking the interleukin-6/gp130 interaction: a two-helix bundle system. Design and conformational studies. AB - The objective of our study was to mimic in a typical reductionist approach the molecular interactions observed at the interface between the gp130 receptor and interleukin-6 during formation of their complex. A peptide system obtained by reproducing some of the interleukin-6/gp130 molecular interactions into a two helix bundle structure was investigated. The solution conformational features of this system were determined by CD and NMR techniques. The CD titration experiments demonstrated that the interaction between the designed peptides is specific and based on a well-defined stoichiometry. The NMR data confirmed some of the structural features of the binding mechanism as predicted by the rational design and indicated that under our conditions the recognition specificity and affinity can be explained by the formation of a two-helix bundle. Thus, the data reported herein represent a promising indication on how to develop new peptides able to interfere with formation of the interleukin-6/gp130 complex. PMID- 12630695 TI - Oligopeptide-mediated helix stabilization of model peptides in aqueous solution. AB - Oligopeptide-mediated helix stabilization of peptides in hydrophobic solutions was previously found by NMR and CD spectroscopic studies. The oligopeptide included the hydrophobic amino acids found in its parent peptide and were interposed by relevant basic oracidic amino acids. The strength of the interactions depended on the amino acid sequences. However, no helix-stabilizing effect was seen for the peptides in phosphate buffer solution, because the peptides assumed a random-coil structure. In order to ascertain whether the helix stabilizing effect of an oligopeptide on its parent peptide could operate in aqueous solution, model peptides EK17 (Ac-AEAAAAEAAAKAAAAKA-NH2) and IFM17 (Ac AEAAAAEIFMKAAAAKA-NH2) that may assume an alpha-helix in aqueous solutions were synthesized. Interactions were examined between various oligopeptides (EAAAK, KAAAE, EIFMK, KIFME, KIFMK and EYYEE) and EK17 or IFM17 in phosphate buffer and in 80% trifluoroethanol (TFE)-20% H2O solutions by CD spectra. EAAAK had little effect on the secondary structures of EK17 in both buffer and TFE solutions, while KAAAE, which has the reverse amino acid sequence of EAAAK, had a marked helix-destabilizing effect on EK17 in TFE. EIFMK and KIFME were found to stabilize the alpha-helical structure of EK17 in phosphate buffer solutions, whereas KIFMK and EYYEE destabilized the alpha-helical structure of EK17. EIFMK and KIFME had no effect on IFM17, because unexpectedly, IFM17 had appreciable amounts of beta-sheet structure in buffer solution. It was concluded that in order for the helix-stabilizing (1) the model peptide, the alpha-helical conformation of which is to be stabilized, should essentially assume an alpha helical structure by nature, and (2) the hydrophobicity of the side-chains of the oligopeptide should be high enough for the oligopeptide to perform stable specific side chain-side chain intermolecular hydrophobic interactions with the model peptide. PMID- 12630696 TI - Isolation of vulgin, a new antifungal polypeptide with mitogenic activity from the pinto bean. AB - An antifungal polypeptide bearing an N-termnial sequence with some homology to chitinases was purified from an extract of pinto beans. The polypeptide, designated vulgin, exerted antifungal activity toward Mycosphaerella arachidicola, Coprinus cornatus, Fusarium oxysporum and Botrytis cinerea. Vulgin inhibited translation in a rabbit reticulocyte lysate system with an IC50 of 4.3 microM and HIV-1 reverse transcriptase activity with an IC50 of 58 microM. Vulgin stimulated in vitro incorporation of methyl [3H] thymidine into mouse splenocytes. PMID- 12630697 TI - Purification and characterization of a new ribonuclease from fruiting bodies of the oyster mushroom Pleurotus ostreatus. AB - A ribonuclease (RNase), possessing an N-terminal sequence disparate from those of ribonucleases from other mushrooms and previously isolated Pleuotus ostreatus RNases, was purified from the fruiting bodies of the edible mushroom Pleurotus ostreatus. The N-terminal sequence of Pleurotus ostreatus RNase did not manifest homology even to a previously reported RNase from the same mushroom. The ribonuclease was adsorbed on CM-Sepharose and Mono S. It exhibited a molecular mass of 12 kDa in both sodium dodecyl sulphate-polyacrylamide gel electrophoresis and gel filtration on Superdex 75. The ribonuclease displayed an activity of 11490 U/mg on yeast tRNA. The highest ribonuclease activity was exhibited toward poly U, followed by poly A and poly C. No activity was shown toward poly G. The optimal pH for its activity was 7 and the optimal temperature was 55 degrees C. It inhibited cell-free translation in a rabbit reticulocyte lysate with an IC50 of 240 nM. PMID- 12630698 TI - Synthesis and circular dichroism study of the human salivary proline-rich protein IB7. AB - The solid phase synthesis of a 59 amino acid human salivary protein IB7 has been accomplished using Fmoc strategy. Because the protein contains 25 proline, 13 glycine and 9 glutamine residues the coupling time, piperidine delivery and acetic anhydride reaction time were increased. Yield after HPLC purification was 35%. Circular dichroism studies revealed that about one third of IB7 residues adopted a type II helix secondary structure, as found in collagen helices. The rest of the sequence adopts a random coil secondary structure. PMID- 12630699 TI - Covalent structure and some pharmacological features of native and cleaved alpha KTx12-1, a four disulfide-bridged toxin from Tityus serrulatus venom. AB - A toxin with four disulfide bridges from Tityus serrulatus venom was able to compete with 125I-kaliotoxin on rat brain synaptosomal preparations, with an IC50 of 46 nM. The obtained amino acid sequence and molecular mass are identical to the previously described butantoxin. Enzymatic cleavages in the native peptide followed by mass spectrometry peptide mapping analysis were used to determine the disulfide bridge pattern of alpha-KTx12-1. Also, after the cleavage of the first six N-terminal residues, including the unusual disulfide bridge which forms an N terminus ring, the potency of the cleaved peptide was found to decrease about 100 fold compared with the native protein. PMID- 12630700 TI - Role of protein kinase C during insulin mediated skeletal muscle cell spreading. AB - Protein kinase C (PKC) is known to play important roles in integrin mediated cell spreading. This study investigated the role of PKC during insulin mediated muscle cell spreading, which was independent of integrin alpha5. We found that PKC-alpha becomes active and localise to membrane during insulin mediated cell spreading. We also found that PKC activation is essential for cell spreading stimulated by insulin and this activation enhances the cell spreading. PKC activation increased the tyrosine phosphorylation of focal adhesion kinase (FAK) and paxillin as well as tyrosine kinase activity of FAK. We also observed that PKC activation enhanced the FAK associated PI 3-kinase activity and also increased the activation of ERK 1/-2. Moreover, the effect of PKC activation on insulin mediated cell spreading as well as tyrosine phosphorylation of FAK and paxillin depends upon integrity of actin cytoskeleton. Thus, PKC is an important signaling protein during insulin mediated muscle cell spreading. PMID- 12630701 TI - The signaling mechanisms of long distance intercellular calcium waves (far waves) in cultured human uterine myocytes. AB - Cultured human myocytes exhibit intercellular calcium waves that travel farther than 100 microm ('far waves'). This work investigates the mechanism of far wave propagation. Culture lines were initiated from myometrial biopsies of term pregnant women. Calcium green-1 was used as a fluorescence probe for intracellular free calcium. Serial imaging was performed at a frame rate of 0.83 frames/s. Intercellular calcium waves were mechanically initiated by atraumatically applying small drops of mineral oil onto the surface of the monolayer. Each intercellular calcium wave was scored using a standardized grid, and points were assigned depending upon the distance the wave traveled and the fluorescent intensity observed within each region. Experiments were performed in the presence of inhibitors of gap junctions and connexin hemichannels (octanol), ATP (apyrase and MDL 12330 A), prostaglandins (indomethacin, high concentrations of lanthanum), the prostaglandin transporter, PGT (DIDS), and transmembrane calcium flux (low concentrations of lanthanum). Octanol, apyrase and MDL 12330 A failed to modify the far waves, indicating gap junctions, connexin hemichannels and ATP do not participate in the paracrine mechanism. Indomethacin at 30, 100 and 300 microM, in a dose dependent manner, reduced the far wave score to 0, suggesting a prostaglandin was critically involved in the mechanism. DIDS reduced the far wave score, but did not fully inhibit wave propagation, suggesting the presence of PGT-dependent and -independent components to the mechanism. Lanthanum at 0.1 mM had no effect, but at 1 mM, reduced the far wave score. These results are consistent with PGF2alpha and/or PGE2 being the signal molecule for the PGT dependent component. Taken together, these data indicate that long distance intercellular calcium waves in cultured human myocytes utilizes a paracrine signaling mechanism, but with more than one extracellular signaling compound. PMID- 12630702 TI - The skeletal muscle ryanodine receptor isoform 1 is found at the intercalated discs in human and mouse hearts. AB - The ryanodine receptors (RyRs) are a class of intracellular calcium release channels of which there are three isoforms. In striated muscle, isoform 1 and isoform 2 are mainly expressed in the terminal cisternae of sarcoplasmic reticulum of skeletal muscle and heart, respectively. Isoform 3 is widely distributed in tissues but in minuscule amounts. These channels release calcium ions from intracellular stores in excitation-contraction coupling for cell signaling. Here, we report the presence of skeletal muscle isoform 1 localized in the intercalated discs (IDs) of human and mouse hearts. By using RyR1 and connexin43 specific antibodies and dual immunofluorescent techniques, both were localized in the proximity of the IDs of human and mouse hearts. We confirmed that RyR1 is localized to the IDs by selective immunoprecipitation of RyR isoform 1 from a subcellular fraction containing IDs from human heart tissue. The functional significance of our observation remains to be elucidated as isoform 1 is involved in depolarization induced calcium release, unlike RyR isoforms 2 and 3 which appear to be involved in calcium induced calcium release. PMID- 12630703 TI - Cardiac neural crest ablation alters aortic smooth muscle force and voltage sensitive Ca2+ responses. AB - Ablation of the premigratory cardiac neural crest (CNC) from the chick embryo results in a malformed outflow tract vasculature termed persistent truncus arteriosus (PTA). In addition, loss of the CNC disrupts myocardial excitation contraction (EC) coupling, decreases intracellular Ca2+ transients, and depresses force generation. We examined if similar defects occurred in the neural crest derived smooth muscle of the aortic arch in a test of the hypothesis that loss of elements from the CNC disrupts EC coupling and force production in the smooth muscle of the tunica media of the aortic arch. Aortic arch segments from chicks (embryonic day 15) displaying PTA generated approximately 43% of stress generated by the aortic arch from sham-operated control embryos during potassium depolarization. The depressed force response was associated with a twofold lower Fura-2 transient. In contrast, force and steady-state Fura-2 signals during endothelin-1 stimulation were unchanged. The differences seen in stress generation with potassium depolarization between sham and PTA displaying embryos were not seen in the descending aorta, a tissue not derived from the neural crest. Protein content and immunostaining revealed no differences in the content of actin, myosin, or dihydropyridine receptor from sham or PTA aortic arch. Our results suggest that the CNC is required for normal aortic arch smooth muscle function and support the hypothesis that the loss of CNC impacts the force generating ability, in part by disruption of the EC-coupling processes and altering Ca(2+)-handling. PMID- 12630704 TI - 2,3-Butanedione monoxime (BDM) as a myosin inhibitor. AB - 2,3-Butanedione monoxime (BDM) is the well-characterized, low-affinity, non competitive inhibitor of skeletal muscle myosin-II. It has been widely used at millimolar concentrations in cell biological experiments with the assumption that it is an ATPase inhibitor of the myosin superfamily. To determine the usefulness of BDM as a general myosin inhibitor, the ATPase activities of the motor domains of skeletal muscle myosin-II, Acanthamoeba myosin-IC, human myole, chicken myosin V, and porcine myosin-VI were measured in the presence of 0-40 mM BDM. BDM inhibits skeletal muscle myosin-II, but it does not inhibit the ATPase activity of the other myosins. Therefore, BDM is not a general inhibitor of the myosin ATPase. BDM has a broad effect on many non-myosin proteins (many uncharacterized), and thus should not be used in whole-cell experiments as a myosin inhibitor. PMID- 12630705 TI - Spectrofluorometric analysis of length-dependent conformational changes in cardiac troponin C. AB - Length modulation of cardiac muscle is manifested in the Frank-Starling relation of the heart. Recently, it has been shown that length-dependent changes in SH reactivity of cardiac troponin C (cTnC) occurred in association with cross-bridge attachment and Ca2+. However, the presence of two SH groups (Cys-35 and Cys-84) in the regulatory region of cTnC complicates efforts to detect conformational changes. In this study skinned porcine cardiac fibers were reacted with 7 diethylamino-3-[4'maleimidylphenyl]-4-methylcoumarin (CPM). Alkaline urea gel electrophoresis, along with protein elution, was used to isolate filament bound cTnC. Analysis of fluorescence measurement showed that there is a Ca(2+) increased fluorescence for CPM-labeled cTnC in long fibers (sarcomere length = 2.2 approximately 2.5 microm) but not in short fibers (sarcomere length = 1.6 approximately 1.8 microm). In addition, the labeled cTnC was measured for the fluorescence decrease over time by adding a non-fluorescence energy acceptor, 4 dimethylaminophenylazophenyl-4'maleimide (DABMI), in the presence and absence of Ca2+. Fluorescence quenching by DABMI is not affected by Ca2+ in long fibers but it is significantly increased in short fibers. However, the fibers maintained in the relaxed state with 5 mM MgATP and 1 mM Vanadate showed no length effect on the CPM-labeled cTnC in terms of the Ca(2+)-mediated changes in fluorescence spectrum and in fluorescence quenching by DABMI. All together, our results suggest that the relative reactivities of Cys-35 and Cys-84 vary with sarcomere length. PMID- 12630706 TI - Human single masseter muscle fibers contain unique combinations of myosin and myosin binding protein C isoforms. AB - Striated craniofacial and limb muscles differ in their embryological origin, regulatory program during myogenesis, and innervation. In an attempt to explore the effects of these differences on the striated muscle phenotype in humans, the expression of myosin and myosin-associated thick filament proteins were studied at the single fiber level both in the human jaw-closing masseter muscle and in two limb muscles (biceps brachii and quadriceps femoris muscles). In the masseter, unique combinations of myosin heavy chain (MyHC) and myosin binding protein C (MyBP-C) isoforms were observed at the single fiber level. Compared to the limb muscles, the MyHC isoform expression was more complex in the masseter while the opposite was observed for MyBP-C. In limb muscles, a coordinated expression of three MyHC and three MyBP-C isoforms were observed, i.e., single fibers contained one or two MyHC isoforms, and up to three MyBP-C isoforms. Also, the relative content of the different MyBP-C isoforms correlated with the MyHC isoform expression. In the masseter, on the other hand, up to five different MyHC isoforms could be observed in the same fiber, but only one MyBP-C isoform was identified irrespective MyHC isoform expression. This MyBP-C isoform had a migration rate similar to the slow MyBP-C isoform in limb muscle fibers. In conclusion, a unique myofibrillar protein isoform expression was observed in the human masseter muscle fibers, suggesting significant differences in structural and functional properties between muscle fibers from human masseter and limb muscles. PMID- 12630707 TI - Separation of detubulation and vacuolation phenomena in amphibian skeletal muscle. AB - Sartorius muscle fibres from cold-adapted Rana temporaria were exposed to variants of an established detubulation procedure (Koutsis et al. (1995) J Muscle Res Cell Motil 16, 519-528) to test the extent to which detubulation and tubular vacuolation phenomena could be separated using different conditions of osmotic shock. A control procedure was optimised to a 28-min exposure to 400 mM glycerol Ringer. This was followed by a recovery step involving its replacement by a Ca2+/Mg(2+)-Ringer solution and steady cooling over 30 min from room temperature (approximately 18 degress C) to approximately 10 degress C, followed by the restoration of the normal Ringer solution. This procedure successfully abolished the action potential after-depolarisation component, reflecting a loss of tubular conduction ('detubulation') in 74.3 +/- 5.9% of the fibres studied. Omitting the cooling during the recovery step sharply reduced the incidence of detubulation. So did omitting either the high-[Ca2+] and/or [Mg2+] in the recovery solutions in test procedures, but to significantly different extents (P < 5%). Yet trapping of fluorescent Sulfhorhodamine B dye in 'closed' vacuoles persisted albeit with reduced proportions of fibre volume occupied by vacuoles. Furthermore, the variations in recovery conditions produced similar levels of vacuolation despite smaller vacuole sizes in the cooled fibres (P < 0.05). These findings demonstrate that fibre vacuolation and detubulation are phenomena that are potentially separable through varying the conditions of osmotic shock, with detubulation requiring significantly more stringent conditions than vacuolation. PMID- 12630708 TI - The effects of physical activity and estrogen treatment on rat fast and slow skeletal muscles following ovariectomy. AB - Decreased estrogen production is associated with changes in the skeletal, cardiovascular and muscular systems. At the level of skeletal muscles, it has been shown that a reduction in force production occurs at menopause but the underlying mechanisms are still unknown. The aim of the study was to investigate the effects of ovariectomy on myosin heavy chain (MyHC) composition. Additionally, we studied the effects of physical activity and the combined effects of physical activity and estrogen treatment on MyHC content in ovariectomised (OX) animals. Twenty-five rats were randomly assigned to five different groups: controls, runners, OX, ovariectomised runners and ovariectomised runners receiving estrogen. Exercise consisted of voluntary running for 5 weeks. Two muscles were analysed: m. extensor digitorum longus, EDL, (fast muscle) and m. soleus (slow muscle). MyHC content was analysed on 8% gel electrophoresis. The level of running activity is reduced in OX animals and estrogen administration is associated with the normalisation of the level of physical activity. Ovariectomy induces a shift from fast to slow MyHC isoforms in both the soleus and EDL. When OX animals are allowed to run, alterations in MyHC isoforms are still observed in the EDL but not in the soleus. When physical activity is combined with estrogen treatment no alterations are observed in both muscles. In conclusion, this study shows that ovariectomy induces alterations in the contractile properties of skeletal muscles and that physical activity in combination with estrogen treatment are associated with the maintenance of slow and fast muscle characteristics. PMID- 12630710 TI - Regulation of striated muscle contraction: a discussion. PMID- 12630711 TI - The Annual Meeting on Muscle Contraction and Cell Motility. Chiba, Japan, 6-8 January 2002. PMID- 12630712 TI - Traditional remedies in lead poisoning. PMID- 12630709 TI - Smooth muscle myosin filament assembly under control of a kinase-related protein (KRP) and caldesmon. AB - Kinase-related protein (KRP) and caldesmon are abundant myosin-binding proteins of smooth muscle. KRP induces the assembly of unphosphorylated smooth muscle myosin filaments in the presence of ATP by promoting the unfolded state of myosin. Based upon electron microscopy data, it was suggested that caldesmon also possessed a KRP-like activity (Katayama et al., 1995, J Biol Chem 270: 3919 3925). However, the nature of its activity remains obscure since caldesmon does not affect the equilibrium between the folded and unfolded state of myosin. Therefore, to gain some insight into this problem we compared the effects of KRP and caldesmon, separately, and together on myosin filaments using turbidity measurements, protein sedimentation and electron microscopy. Turbidity assays demonstrated that KRP reduced myosin filament aggregation, while caldesmon had no effect. Additionally, neither caldesmon nor its N-terminal myosin binding domain (N152) induced myosin polymerization at subthreshold Mg2+ concentrations in the presence of ATP, whereas the filament promoting action of KRP was enhanced by Mg2+. Moreover, the amino-terminal myosin binding fragment of caldesmon, like the whole protein, antagonizes Mg(2+)-induced myosin filament formation. In electron microscopy experiments, caldesmon shortened myosin filaments in the presence of Mg2+ and KRP, but N152 failed to change their appearance from control. Therefore, the primary distinction between caldesmon and KRP appears to be that caldesmon interacts with myosin to limit filament extension, while KRP induces filament propagation into defined polymers. Transfection of tagged-KRP into fibroblasts and overlay of fibroblast cytoskeletons with Cy3KRP demonstrated that KRP colocalizes with myosin structures in vivo. We propose a new model that through their independent binding to myosin and differential effects on myosin dynamics, caldesmon and KRP can, in concert, control the length and polymerization state of myosin filaments. PMID- 12630713 TI - Growth of infants during the first year of life according to feeding regimen in the first 4 months. AB - This study aimed to compare the first-year growth of infants who had received different feeding regimens throughout the first 4 months. Anthropometric measurements of 332 infants attending a well child clinic were analysed. The infants were divided into four groups: exclusively breastfed (BF), predominantly BF, partially BF, and non-BE Exclusively BF infants were significantly heavier in the first 2 months of life compared to partially BF or non-BF infants Weight and length measurements of the predominantly BF infants were almost identical to those of the exclusively BF group at all ages. Our data show that the exclusively BF infants have a different growth pattern during the first year of life than those of partially BF or non-BF infants. PMID- 12630714 TI - Cerebrospinal fluid lysozyme level for the diagnosis of tuberculous meningitis in children. AB - Lysozyme activity was assayed in the cerebrospinal fluid (CSF) of 32 tuberculous meningitis (TBM), 17 bacterial meningitis, 10 partially treated bacterial meningitis, 18 encephalitis and 18 control subjects. The mean CSF lysozyme activity was significantly raised (p < 0.001) in TBM patients compared with other study groups. A cut-off CSF lysozyme level of > or = 26 U/l had a sensitivity and specificity of 93.7 and 84.1 per cent, respectively for the diagnosis of TBM. Overall, it was found to be a better test than any other single test and thus can be used for rapid and early diagnosis of TBM in children. PMID- 12630716 TI - Seasonal dietary stress in Peruvian children. AB - This article examines the seasonal variation in energy intake among young children in a fanning community in southern Peru. Caloric intakes among young children fall to their lowest annual level during the post-harvest season even though food and cash resources are at their highest levels. Among toddlers (1-3 years) the decline is statistically significant (p = 0.006). Their energy intake meets only 65.6 per cent of their predicted requirement during the post-harvest season. In contrast to the literature, dietary stress for young children is not greatest in the pre-harvest season. Also, household food availability does not accurately indicate dietary risk among young children in this community. PMID- 12630715 TI - Predictors and nutritional consequences of intestinal parasitic infections in rural Ecuadorian children. AB - The study investigated the prevalence, risk factors, and nutritional consequences of intestinal parasitic infections (IPI) in rural Ecuadorian children. A total of 244 children aged 0.2-14 years were studied. The data were collected using a structured questionnaire, anthropometry, and laboratory analysis of blood and fecal samples. The results showed that 90 per cent of the subjects were infected with at least one pathogenic IPI: 51 per cent with helminths, 37.6 per cent with protozoa, and 21.4 per cent with both. Giardia-infected children had a risk for stunted growth that was twice that of other children (51.7 vs. 33.1 per cent; OR = 2.16, 95 per cent CI = 1.13-4.15; p = 0.01). They also had significantly reduced mean hemoglobin levels compared with their non-infected counterparts (11.8 +/- 1.5 vs. 12.2 +/- 1.4 g/dl; p = 0.023). However, the proportion diagnosed with iron-deficiency anemia was slightly, but not significantly, increased (29.4 vs. 24.3 per cent). The most consistent predictor of Giardia and other protozoal IPI risk was a high intra-/peri-domicilliary concentration of domestic animals. Children who lived in such households had an infection risk that was two to five times greater than that of their non-infected counterparts. The data indicate that Giardia intestinalis infection has an adverse impact on child linear growth and hemoglobin. They also suggest that domestic animals may be an important reservoir for Giardia and other intestinal protozoal infections observed in the Ecuadorian children studied. PMID- 12630717 TI - Percentage, bacterial etiology and antibiotic susceptibility of acute respiratory infection and pneumonia among children in rural Senegal. AB - Acute respiratory infections (ARI) are still a major health problem in most developing countries. So far no study has evaluated the importance of childhood ARI in rural Senegal. We prospectively studied ARI, the percentage of pneumonia and related mortality, as well as the bacterial composition of nasopharyngeal flora using nasopharyngeal aspirates in 114 children, aged 2-59 months, presenting at Ndioum's pediatric ward. Excluded from the trial were those children that had had antimicrobial therapy in the previous 2 weeks. The Kirby Bauer method was used to determine antibiotic resistance throughout the study. The percentage of ARI and pneumonia among the population tested was 24 per cent and 11 per cent respectively. Streptococcus pneumonia was often resistant to cotrimoxazole (31 per cent) but only 9 per cent were resistant to chloramphenicol and 14 per cent to penicillin. Haemophilus influenzae (HI) was uniformly sensitive to ampicillin, and only 4 per cent were resistant to chloramphenicol and 11 per cent to cotrimoxazole. We conclude that SP and HI resistance to cotrimoxazole is important and warrants larger clinical trials using chloramphenicol. Information campaigns and intense management of comorbidities are desirable in this type of population. Comorbidities (tuberculosis, malaria, HIV-AIDS, severe malnutrition) are determinant variables in many ARI cases and carry a high negative prognosis value. PMID- 12630718 TI - The role of nitric oxide in pediatric patients with portal hypertension. AB - The hyperdynamic circulation of cirrhosis and portal hypertension has been postulated to be due to the vasodilatory effects of nitric oxide. However, there have been conflicting results in adults and no studies in children. We aimed to measure the nitric oxide level in serum of pediatric patients with portal hypertension with and without cirrhosis, in order to assess its role in the development of hemodynamic changes. We measured nitric oxide levels in 41 pediatric patients (21 patients with intrahepatic portal hypertension and 20 with extrahepatic portal hypertension). The mean age of the study population was 11.2 +/- 4.6 years; 53 per cent were female. Twenty healthy children were included as a control group. Nitric oxide levels were measured by Boehringer-Mannheim colorimetric assay and the statistical significance was calculated by Kruskal Wallis one-way ANOVA. Significantly higher nitric oxide levels were found in patients independent of the type of portal hypertension compared with the control group (29.4 +/- 6 in patients with intrahepatic portal hypertension, 29.5 +/- 5.8 in patients with extrahepatic portal hypertension, and 23.6 +/- 6.5 in the control group; p < 0.007). These data showed a difference between the groups and suggest that nitric oxide, predominantly independent of cirrhosis, plays a primary role in the pathogenesis of portal hypertension. PMID- 12630720 TI - Vitamin A status and nutritional intake of carotenoids of preschool children in Ijaye Orile community in Nigeria. AB - This study was carried out to determine the vitamin A status and nutritional intake of carotenoids of 213 children between the ages of 6 months and 6 years in a rural community in Nigeria. There were 109 males and 104 females. A total of 57 (26.8 per cent) children were deficient in serum retinol levels (< 10 microg/dl) while 102 (47.9 per cent) had low levels (10-19 microg/dl). The highest prevalence of serum retinol deficiency was in the 6-12 months age group, most of whom were breastfeeding and there was poor correlation between duration of breastfeeding and serum retinol levels. The prevalence of night-blindness was 1.5 per cent; however, none of the children had xerophthalmia. Chronically malnourished children had lower mean serum retinol levels than well nourished children. There was a high consumption of carotenoid-containing food, but despite this there was a high prevalence of vitamin A deficiency. We therefore suggest that measures to combat vitamin A deficiency should include vitamin A supplementation on a short-term basis. On a long-term basis parents should be educated on the importance of the consumption of locally available sources of provitamin A and pre-formed vitamin A rich foods, and the avoidance of overcooking. Parents should also be encouraged to grow more beta-carotene containing foods. PMID- 12630719 TI - Antimicrobial susceptibility and distribution of non-typhoidal Salmonella serovars isolated in Malaysian children. AB - There is widespread resistance of Salmonella species to commonly prescribed antimicrobials the world over. We aimed to determine the antimicrobial susceptibility and serovar distribution of non-typhoidal Salmonella (NTS) isolated from blood cultures of Malaysian children. Positive isolates of NTS from blood cultures obtained from children admitted to the pediatric wards of University of Malaya Medical Center (UMMC), a large urban hospital from Kuala Lumpur (1991-2001), and Hospital Kota Bharu (HKB), from the predominantly rural state of Kelantan (1991-1999), Malaysia, were reviewed retrospectively. Serovar distribution and antimicrobial susceptibility were ascertained. A total of 64 and 55 isolates of NTS were obtained from blood cultures of children admitted to UMMC and HKB, respectively. The commonest serovar isolated was Salmonella enteritidis in both centers. The NTS isolated were highly sensitive to the antimicrobials tested: ampicillin 98 per cent, chloramphenicol 98 per cent, gentamicin 97 per cent, trimethoprim-sulfamethoxazole (TMP-SMX) 98 per cent, and ceftriaxone 100 per cent in UMMC; ampicillin 100 per cent, chloramphenicol 87 per cent, kanamycin 100 per cent, streptomycin 96 per cent, TMP-SMX 93 per cent, and tetracycline 89 per cent in HKB. There were only one and five multi-resistant isolates in UMMC and HKB, respectively. In conclusion, NTS isolated from blood cultures of Malaysian children from Kuala Lumpur and Kota Bharu were highly sensitive to commonly prescribed antibiotics. We speculate that this is due to the restriction of sales of antimicrobials in Malaysia except by prescription. Continuing vigilance and frequent antmicrobial surveillance is necessary. PMID- 12630721 TI - Japanese encephalitis in and around Pondicherry, South India: a clinical appraisal and prognostic indicators for the outcome. AB - Japanese encephalitis (JE) is numerically one of the most important causes of viral encephalitis worldwide, with an estimated 50,000 cases and 15,000 deaths annually. About one-third of patients die and half of the survivors have severe neuropsychiatric sequelae. Three hundred patients clinically suspected of JE were tested in the present study. Laboratory confirmation of JE was on the basis of detection of antigen or presence of JE-specific IgM antibody and/or neutralizing antibody in a single CSF sample. The risk factors that were associated with fatal outcome were determined. Japanese encephalitis infection was confirmed in 70.7 per cent (212/300) of the patients. All patients were from rural areas and with low socioeconomic background. Prominent clinical findings were: fever in 100 per cent (212/212) patients, altered sensorium in 87.73 per cent (186/212), convulsion in 85.84 per cent (182/212), headache in 50 per cent (106/212), and vomiting in 47.64 per cent (101/212). The final clinical outcome was available for only 68.39 per cent (145/212) of patients, as children were taken home against medical advice. Of these, 35.86 per cent (52) died while 63.44 per cent (92) of patients survived. Correlations of investigative findings with the final outcome revealed that absence of virus-specific IgM and neutralizing antibodies in CSF were associated with fatal outcome. In patients diagnosed with Japanese encephalitis the presence of a virus-specific immune response is associated with a favourable outcome and an important parameter in recovery from illness. PMID- 12630722 TI - Cystic fibrosis mutation I1234V in a Qatari lady. AB - We describe a late diagnosis of cystic fibrosis (CF) in a multiparous Qatari lady, in whom the main presenting symptoms were those of chronic lung disease. Genetic analysis showed that the patient has a homozygous mutation I1234V in the cystic fibrosis transmembrane conductance regulator gene. This suggests that this mutation has a variable expression of clinical severity and long survival. PMID- 12630723 TI - Calcaneal apophysitis due to brucellosis. AB - Brucellosis is still a major health problem in many geographical areas. Osteoarticular complications are important owing to their high prevalence. We report an unusual case of childhood brucellosis presenting with septic apophysitis of the calcaneus and abscess formation in a 12-year-old boy. The patient was successfully treated with a combination of antibiotics. The importance of early recognition of the disease and differential diagnosis is emphasized. Early recognition of infection, prolonged treatment, and long-term follow-up may improve the outcome. PMID- 12630725 TI - Pneumonia associated with visceral leishmaniasis in childhood. PMID- 12630724 TI - First case of neurocysticercosis in Saudi Arabia. PMID- 12630726 TI - Surfactant therapy in the absence of respiratory support facilities. PMID- 12630727 TI - Plasma chitotriosidase activity in acute Plasmodium falciparum malaria. PMID- 12630728 TI - The importance of a high index of suspicion in rabies. PMID- 12630729 TI - Emergency airway management. AB - Airway control is one of the most critical interventions required for saving a life. It is essential that practitioners be as well trained as possible in the numerous techniques available to establish airway control. This article reviews some of the available techniques, though other techniques that are not discussed (such as fiberoptic-assisted endotracheal intubation) may also be useful. Perhaps the most important aspect of advanced airway management is the ability to anticipate and prepare for the difficult airway. This article gives numerous options for the difficult airway situation. PMID- 12630730 TI - Diagnosis of acute cardiac ischemia. AB - A better understanding of coronary syndromes allow physicians to appreciate UAP and AMI as part of a continuum of ACI. ACI is a life-threatening condition whose identification can have major economic and therapeutic importance as far as threatening dysrhythmias and preventing or limiting myocardial infarction size. The identification of ACI continues to challenge the skill of even experienced clinicians, yet physicians continue (appropriately) to admit the overwhelming majority of patients with ACI; in the process, they admit many patients without acute ischemia [2], overestimating the likelihood of ischemia in low-risk patients because of magnified concern for this diagnosis for prognostic and therapeutic reasons. Studies of admitting practices from a decade ago have yielded useful clinical information but have shown that neither clinical symptoms nor the ECG could reliably distinguish most patients with ACI from those with other conditions. Most studies have evaluated the accuracy of various technologies for diagnosing ACI, yet only a few have evaluated the clinical impact of routine use. The prehospital 12-lead ECG has moderate sensitivity and specificity for the diagnosis of ACI. It has demonstrated a reduction of the mean time to thrombolysis by 33 minutes and short-term overall mortality in randomized trials. In the general ED setting, only the ACI-TIPI has demonstrated, in a large scale multicenter clinical trial, a reduction in unnecessary hospitalizations without decreasing the rate of appropriate admission for patients with ACI. The Goldman chest pain protocol has good sensitivity for AMI but was not shown to result in any differences in hospitalization rate, length of stay, or estimated costs in the single clinical impact study performed. The protocol's applicability to patients with UAP has not been evaluated. Single measurement of biomarkers at presentation to the ED has poor sensitivity for AMI, although most biomarkers have high specificity. Serial measurements can greatly increase the sensitivity for AMI while maintaining their excellent specificity. Biomarkers cannot identify most patients with UAP. Finally, diagnostic technologies to evaluate ACI in selected populations, such as echocardiography, sestamibi perfusion imaging, and stress ECG, may have very good to excellent sensitivity; however, they have not been sufficiently studied. PMID- 12630731 TI - Pearls and pitfalls in the emergency department evaluation of abdominal pain. AB - Abdominal pain is a common complaint in the emergency department (ED), comprising approximately 5% of total visits. Although not typically serious, abdominal pain often presents many difficult situations for the clinician. Women of reproductive age and elderly individuals represent some of the most challenging patients to evaluate. Although there are general diagnostic and clinical principles that apply to the evaluation of all patients, these two groups deserve extra attention because of the broad differential diagnosis and potential for serious complications. This article outlines the epidemiology of abdominal pain in patients who present to the ED, and provides guidelines pertinent to the evaluation of all patients. In addition, the article discusses the approach to abdominal pain of pelvic origin in women of reproductive age and abdominal pain in elderly individuals. PMID- 12630732 TI - Diagnosis of subarachnoid hemorrhage in the emergency department. AB - To decide which patients with headache ought to be evaluated for SAH, physicians should focus on specific elements of the patient history, such as onset, severity, and quality of the headache and associated symptoms. These questions should be asked and the responses documented for every patient with a headache. The physical examination should be compulsive with regard to vital signs, HEENT. and neurologic signs. Then, the physician should form an explicit differential diagnosis and have reasons for diagnosing migraine, tension, or sinus headache and other benign causes. If there is no clear-cut alternative hypothesis, the patient should be evaluated by CT and LP (if the CT is negative, equivocal, or technically inadequate). Physicians should understand the limitations of this diagnostic algorithm. In addition, the CSF should be carefully analyzed, including measuring the opening pressure. In patients whose CT scans and CSF analyses are normal, further testing is rarely indicated. PMID- 12630733 TI - Evaluation and management of febrile infants in the emergency department. AB - Managing fever in pediatric patients has always been a high-risk aspect of emergency medicine, and this is especially true of infants. The task of correctly discerning "well" versus "ill" can be difficult for these patients because they cannot describe their symptoms and have not developed normal social behaviors. Recent studies, however, have greatly enhanced physicians' understanding of how best to manage these young patients. With this research comes an array of new recommendations for diagnosing and treating febrile infants, which can be incorporated into the practice of every emergency physician who cares for pediatric patients. PMID- 12630734 TI - Management of toxic exposure in children. AB - Millions of children ingest household products and medications yearly. The continuous proliferation of new products and pharmaceutic agents makes it difficult for physicians to maintain a current command of toxicologic information. Multiple sources, including poison control centers, can provide information; however, EPs must be familiar with several agents that are either significant for their frequency or for their disproportionate potential for morbidity and mortality in pediatric patients. With this select group of intoxicants, physicians must anticipate cardiovascular and pulmonary instability and rapid changes in central nervous system functioning. Appropriate supportive care requires monitoring of the following: vital signs, level of consciousness, airway control, ventilation and circulatory support, body temperature, urine output, and acid base balance. Once these concerns are addressed, prevention of further absorption, enhancing a product's elimination, and treatment with specific antidotes may enhance supportive care. Care is also likely to be enhanced if the EP recognizes the inherent differences (medically and socially) between adults and children of various ages. Definitive emergency care is completed only after the provision of a developmentally oriented preventive strategy. PMID- 12630735 TI - Procedural sedation and analgesia for children in the emergency department. AB - PSA for children continues to be an integral part of the practice of emergency medicine. The advancement of knowledge for all health care professionals must continue so that pain and anxiety in children may be minimized during their visits to EDs. PMID- 12630736 TI - Medication errors in the emergency department: a systems approach to minimizing risk. AB - Adverse drug events caused by medication errors represent a common cause of patient injury in the practice of medicine. Many medication errors are preventable and hence particularly tragic when they occur, often with serious consequences. The enormous increase in the number of available drugs on the market makes it all but impossible for physicians, nurses, and pharmacists to possess the knowledge base necessary for fail-safe medication practice. Indeed, the greatest single systemic factor associated with medication errors is a deficiency in the knowledge requisite to the safe use of drugs. It is vital that physicians, nurses, and pharmacists have at their immediate disposal up-to-date drug references. Patients presenting for care in EDs are usually unfamiliar to their EPs and nurses, and the unique patient factors affecting medication response and toxicity are obscured. An appropriate history, physical examination, and diagnostic workup will assist EPs, nurses, and pharmacists in selecting the safest and most optimum therapeutic regimen for each patient. EDs deliver care "24/7" and are open when valuable information resources, such as hospital pharmacists and previously treating physicians, may not be available for consultation. A systems approach to the complex problem of medication errors will help emergency clinicians eliminate preventable adverse drug events and achieve a goal of a zero-defects system, in which medication errors are a thing of the past. New developments in information technology and the advent of electronic medical records with computerized physician order entry, ward-based clinical pharmacists, and standardized bar codes promise substantial reductions in the incidence of medication errors and adverse drug events. ED patients expect and deserve nothing less than the safest possible emergency medicine service. PMID- 12630738 TI - Management of lacerations in the emergency department. AB - The goals of wound management are simple: avoid infection and achieve a functional and aesthetically pleasing scar. This is achieved by reducing tissue contamination, debriding devitalized tissue, and restoring perfusion in poorly perfused wounds, in conjunction with a well-approximated skin closure. PMID- 12630737 TI - Evaluation and management of the high-risk orthopedic emergency. AB - Orthopedic injuries are frequently seen in the ED. Whereas the diagnosis and management of most of these injuries is straightforward, there are distinct pitfalls to avoid. The common theme among the high-risk "pitfall" injuries discussed in this article, besides a thorough patient history and careful physical examination with appropriate radiographs, is a high suspicion for the presence of these injuries. When the EP is knowledgeable about these orthopedic pitfalls, these injuries are much less likely to slip by in clinical practice. PMID- 12630739 TI - Correlation between near-infrared spectroscopy and magnetic resonance imaging of rat brain oxygenation modulation. AB - We measure the tissue oxygen and haemoglobin concentrations in the rat brain during modulation of inhaled oxygen concentration (FiO2), using non-invasive frequency domain near-infrared oximetry. The rise in oxygenated haemoglobin concentration and the decline in deoxygenated haemoglobin concentration are demonstrated in correspondence with the modulation of FiO2, which is changed from 20% to 100% in increments of 20%. Furthermore, the tissue oxygenation saturation also shows the corresponding trend and changes ranging from approximately 70% to 90%. The relative changes in deoxygenated haemoglobin concentration are compared to the blood-oxygenation-level-dependent (BOLD) MRI signal recorded during a similar FiO2 protocol. A linear relationship with high correlation coefficient between the relative changes in the BOLD MRI signal and the NIRS signal is observed. PMID- 12630740 TI - Characterization of a single LSO crystal layer high resolution research tomograph. AB - The purpose of this study was to determine the performance of a single lutetium oxy-orthosilicate (LSO) crystal layer High Resolution Research Tomograph (HRRT) positron emission tomography (PET) scanner. The HRRT is a high resolution PET scanner designed for human brain and small animal imaging. The scanner consists of eight panel detectors, which have one layer of 2.1 x 2.1 x 7.5 mm thick LSO crystals. Several phantom studies were performed to determine scanner characteristics, such as resolution, scatter fraction, count rate and noise equivalent count rates (NECR). NECR curves were measured according to both NEMA NU2-1994 and NU2-2001 for three different energy windows, i.e. lower level discriminators (lld) of 350, 400 and 450 keV and an upper level discriminator (uld) of 650 keV. Accuracy of scatter and single photon attenuation corrections was evaluated according to NU2-1994. Data were acquired using a ring difference of 67 and a span of 9. Reconstructions were performed using FORE + 2D FBP or OSEM. Transaxial resolution varied from 2.7 to 2.9 mm FWHM between I and 10 cm off centre locations, and axial resolution varied from 3.2 to 4.4 mm FWHM. Scatter fractions (NU2-1994) equalled 0.31, 0.42 and 0.54 for lld of 450, 400 and 350 keV, respectively. NECR data were highest for an lid of 400 keV and showed a maximum of 46 kcps at 38 kBq cm(-3). Lower NECR values were observed according to NU2-2001, but were still optimal for an lld of 400 keV. After scatter and attenuation corrections, pixel values within water, air and teflon inserts of the NU2-1994 phantom were 14, 4 and 35% of the background activity, respectively. The single layer LSO HRRT scanner shows excellent spatial resolution, making it suitable for small animal studies. The low count rate performance, due to the small amount of LSO, prohibits studies of the human brain, but is sufficient for studies in small laboratory animals. PMID- 12630741 TI - An adaptive algorithm for the detection of microcalcifications in simulated low dose mammography. AB - This paper uses the task of microcalcification detection as a benchmark problem to assess the potential for dose reduction in x-ray mammography. We present the results of a newly developed algorithm for detection of microcalcifications as a case study for a typical commercial film-screen system (Kodak Min-R 2000/2190). The first part of the paper deals with the simulation of dose reduction for film screen mammography based on a physical model of the imaging process. Use of a more sensitive film-screen system is expected to result in additional smoothing of the image. We introduce two different models of that behaviour, called moderate and strong smoothing. We then present an adaptive, model-based microcalcification detection algorithm. Comparing detection results with ground truth images obtained under the supervision of an expert radiologist allows us to establish the soundness of the detection algorithm. We measure the performance on the dose-reduced images in order to assess the loss of information due to dose reduction. It turns out that the smoothing behaviour has a strong influence on detection rates. For moderate smoothing. a dose reduction by 25% has no serious influence on the detection results. whereas a dose reduction by 50% already entails a marked deterioration of the performance. Strong smoothing generally leads to an unacceptable loss of image quality. The test results emphasize the impact of the more sensitive film-screen system and its characteristics on the problem of assessing the potential for dose reduction in film-screen mammography. The general approach presented in the paper can be adapted to fully digital mammography. PMID- 12630742 TI - A new approach to elastography using mutual information and finite elements. AB - Historically, increased mechanical stiffness during tissue palpation exams has been associated with assessing organ health as well as with detecting the growth of a potentially life-threatening cell mass. As such, techniques to image elasticity parameters (i.e., elastography) have recently become of great interest to scientists. In this work, a new method of elastography will be introduced within the context of mammographic imaging. The elastography method proposed represents a non-rigid iterative image registration algorithm that varies material properties within a finite element model to improve registration. More specifically, regional measures of image similarity are used within an objective function minimization framework to reconstruct elasticity images of tissue stiffness. Numerical simulations illustrate: (1) the encoding of stiffness information within the context of a regional image similarity criterion, (2) the methodology for an iterative elastographic imaging framework and (3) elasticity reconstruction simulations. The real strength in this approach is that images from any modality (e.g., magnetic resonance, computed tomography, ultrasound. etc) that have sufficient anatomically-based intensity heterogeneity and remain consistent from a pre- to a post-deformed state could be used in this paradigm. PMID- 12630743 TI - A method for generating patient-specific finite element meshes for head modelling. AB - Finite element modelling of fields within the body, whether electrical or optical, requires knowledge of the geometry of the object being examined. It can be clinically impractical to obtain accurate surface information for individual patients, although a limited set of measurements such as the locations of sensors attached to the body, can be acquired more readily. In this paper, we describe how a generic surface taken from an adult head is warped to fit points measured on a neonatal head surface to provide a new, individual surface from which a finite element mesh was generated. Simulations show that data generated from this mesh and from the original neonatal head surface are similar to within experimental errors. However, data generated from a mesh of the best fit sphere were significantly different from data generated from the original neonatal head surface. PMID- 12630744 TI - Three-dimensional dose verification with x-ray films in conformal carbon ion therapy. AB - A three-dimensional dose verification with photographic emulsions (x-ray films) was realized within the tumour therapy project at GSI Darmstadt, using carbon ions. We present Bragg-peak measurements for 88, 100 and 200 MeV/u carbon ion beams as the simplest case of dose verification. An actual patient treatment plan, composed by the superposition of Bragg-peaks having different energies and intensities, was used to perform a three-dimensional dose verification of an irregularly formed target volume. The shape of the measured dose distribution from film closely matches the intended irradiation volume. Furthermore, the calculated and measured optical density distribution is in good agreement with a maximum deviation of less than 10%. PMID- 12630746 TI - A precorrected-fFT method to accelerate the solution of the forward problem in magnetoencephalography. AB - Accurate localization of brain activity recorded by magnetoencephalography (MEG) requires that the forward problem, i.e. the magnetic field caused by a dipolar source current in a homogeneous volume conductor, be solved precisely. We have used the Galerkin method with piecewise linear basis functions in the boundary element method to improve the solution of the forward problem. In addition, we have replaced the direct method, i.e. the LU decomposition, by a modern iterative method to solve the dense linear system of equations arising from the boundary element discretization. In this paper we describe a precorrected-FFT method which we have combined with the iterative method to accelerate the solution of the forward problem and to avoid the explicit formation of the dense coefficient matrix. For example, with a triangular mesh of 18,000 triangles, the CPU time to solve the forward problem was decreased from 3.5 h to less than 5 min, and the computer memory requirements were decreased from 1.3 GB to 156 MB. The method makes it possible to solve quickly significantly larger problems with widely-used workstations. PMID- 12630745 TI - The influence of brachytherapy dose heterogeneity on estimates of alpha/beta for prostate cancer. AB - The sensitivity of estimates of alpha/beta for prostate tumours to dose heterogeneity in 125I brachytherapy implants, as well as to variation in selected radiobiological parameters, is analysed. The tumour control probabilities of brachytherapy and external beam radiotherapy are equated for ranges of alpha, Tpot, RBE and external beam dose. For each combination of parameters, the equality is used to derive the value of alpha/beta. Different clinical (non uniform) brachytherapy dose distributions, and three uniform brachytherapy dose distributions (120, 144 and 160 Gy) are used. For 'nominal' input parameter values of Tpot = 45 days, alpha = 0.2 Gy(-1), RBE = 1.4, and an external beam dose of 70 Gy, the values obtained for alpha/beta ranged between 2.1 and 12.3 Gy for all of the clinical DVHs, between 2.1 and 3.8 Gy for the better quality clinical implants and between 1.0 and 1.8 Gy for the uniform brachytherapy doses. When only 2% of the volume receiving the lowest dose is omitted from the clinical DVHs, the estimated alpha/beta values ranged between 1.4 and 2.1 Gy. When ranges of input parameters were also considered, the overall range of alpha/beta values for the clinical brachytherapy dose distributions lay between 1.1 and 12.3 Gy for the three best clinical implants, and between 0.7 and 6.3 Gy for uniform doses. We conclude that estimation of alpha/beta without taking into account dose heterogeneity and inter-patient variation may underestimate the actual value alpha/beta. PMID- 12630747 TI - Models and measurements of light intensity changes during laser interstitial thermal therapy: implications for optical monitoring of the coagulation boundary location. AB - We have developed a multi-region spherical Monte Carlo (MC) model to simulate the dynamic changes in light intensity measured during laser interstitial thermal therapy (LITT). Model predictions were validated experimentally in tissue simulating albumen phantoms with well-characterized optical properties that vary dynamically with LITT in a way similar to tissue. For long treatments (2.5 W, approximately 1800 s), the transient light intensity changes demonstrated better qualitative agreement with a three-region MC model (with an inner layer of fully coagulated optical properties, a middle layer of partially coagulated properties and an outer region of native properties); for short treatments (4 W, approximately 240 s), better qualitative agreement was seen with a two-region MC model (with an inner layer of fully coagulated properties and outer region of native properties). These differences were attributed to differences in coagulation formation during low- and high-powered heating regimes, respectively. At the end of heating, a three-region coagulation zone was observed for both heating schemes. Quantitatively, final light intensity changes at the end of heating were compared with changes predicted by both two- and three-region MC for the same experimentally measured coagulation size and found to agree within approximately 30% for both models. The developed MC model helps lend insight into the nature of thermal coagulation events occurring for low and high power LITT irradiation schemes. PMID- 12630748 TI - The direct measurement using an imaging plate for coincidence of radiation centre and laser position in external radiation therapy. AB - A new method of quality assurance has been studied to measure coincidence of the radiation centre and a patient-setup laser position on a transverse plane to the beam at the isocentre. This measurement is achieved by using an imaging plate (IP). When radiation is applied to an IP, the energy is stored as trapped electrons. The number of electrons is decreased by local laser exposure. As a result, the radiation field produced by external beam irradiation is recorded as 'positive' information and the position of the patient-setup laser is recorded as 'negative' on an IP. The advantages of this method are the direct measurement, short time and high resolution. These are required for daily and monthly quality checks. We confirmed the advantage of this method by an experiment using a proton beam. PMID- 12630749 TI - Reply to 'Comment on 'A systematic review of the precision and accuracy of dose measurements in photon radiotherapy using polymer and Fricke MRI gel dosimetry''. PMID- 12630750 TI - DNA methylation in systemic lupus erythematosus. AB - Recent studies on epigenetics, including DNA methylation and its regulatory enzymes, seem likely to contribute to elucidation of the pathogenesis of autoimmune diseases such as systemic lupus erythematosus (SLE), although the relationship between DNA methylation and SLE has long been the subject of investigation. To obtain a deeper understanding of the role of DNA methylation in the induction of SLE, we reviewed the relationship between DNA methylation and SLE based on findings reported in the literature and our own data. Various studies, including ours, have indicated the possible importance of DNA methylation, especially hypomethylation, in the etiology of SLE. These epigenetic studies may give us clues towards elucidation of the pathogenesis of SLE and development of new therapeutic strategies for this disease. PMID- 12630751 TI - The transforming growth factor-beta1 gene polymorphism (G915C) is not associated with systemic lupus erythematosus. AB - Lymphocyte production of transforming growth factor (TGF)-beta1 is decreased in systemic lupus erythematosus (SLE). The lack of this immunoregulatory cytokine may contribute to the characteristic T cell disregulation and aberrant B cell stimulation in SLE patients. The less common C allele of the TGFB1 polymorphism (G915C) is associated with a lower TGF-beta1 production capacity. We performed a population-based case-control study to analyse the impact of this polymorphism on disease susceptibility, on clinical SLE manifestations and autoantibody production. A total of 203 German Caucasian SLE patients (fulfilling the 1982 ACR disease duration 11.5 +/- 7.0 years) and 158 ethnically, age- and sex-matched healthy controls were genotyped with a mutagenically separated polymerase chain reaction. There were no significant differences in the genotype distribution and allele frequencies between patients (915 C = 0.08) and healthy controls (915 C = 0.10). Comparing subgroups of patients, we found no association of major disease manifestations or specific autoantibodies with TGFB1 genotypes or alleles. The TGFB1 polymorphism (G915C) neither significantly contributes to the disease susceptibility, nor predisposes to clinical and immunological manifestations typical of SLE. Further studies are needed to corroborate the pathogenic role of TGF-beta1 in SLE patients and to identify the precise genetic elements controlling its production. PMID- 12630752 TI - Neurological involvement as a poor prognostic factor in catastrophic antiphospholipid syndrome: autopsy findings in 12 cases. AB - CAPS is an uncommon disease, characterized by clinical evidence of multiple organ involvement and histopathological evidence of multiple vessel occlusions, in patients with either primary or secondary antiphospholipid syndrome. The present series describes the clinical manifestations and autopsy findings of 12 patients with CAPS. Neurological involvement was considered the main cause of death in all of them. CNS pathology revealed thrombotic microangiopathy as well as small and large vessel occlusions in several brain areas. Neurological involvement in CAPS is strongly associated with thrombotic microangiopathy and should be considered a potential cause of death in these patients. PMID- 12630753 TI - Rapid clinical progression to diagnosis among African-American men with systemic lupus erythematosus. AB - The initial clinical course of systemic lupus erythematosus (SLE) is variable, ranging from relatively minor manifestations progressing over years to rapid onset of fulminate disease. We sought to identify factors associated with the rapid manifestation of SLE. Chart review of military medical records was used to identify 130 patients who met the American College of Rheumatology classification criteria for SLE. Demographics, clinical criteria date of occurrence, and the date of SLE classification (at least four clinical criteria) met were documented. Prospectively stored serum samples prior to the diagnosis were evaluated for SLE autoantibodies. Median time from the first recorded criteria to diagnosis was significantly shorter in African-American (AA) males compared with AA females and European American (EA) females and males combined. AA males were more likely to have nephritis as their first clinical symptom. Also, less time transpired between the first clinical criterion and SLE diagnosis in AA males with nephritis than in other groups presenting with nephritis. Even when cases presenting with nephritis were excluded, a diagnosis of SLE was made more rapidly in AA males. African-American men progress from initial clinical manifestations to SLE diagnosis more rapidly than other ethnic or gender groups. PMID- 12630754 TI - Lack of association between hyperprolactinemia and soluble IL-2 receptor levels in systemic lupus erythematosus. AB - The aim of the study was to evaluate the association between hyperprolactinemia and T lymphocyte activation through the soluble IL-2 receptor (sIL-2R) in systemic lupus erythematosus (SLE) patients. Seventy SLE patients, 18 of them with hyperprolactinemia (HPRL), were compared with 18 normoprolactinemic (NPRL) patients and 10 age-matched healthy blood-bank donor women. Patients were evaluated by means of the SLE activity index (SLEDAI). Total serum IgG and sIL-2R levels were determined by an ELISA assay. Differences between sIL-2R and IgG serum levels in patients and controls were examined by Kruskal-Wallis analysis and a Spearman r correlation to determine the association between sIL-2R, IgG and prolactin (PRL) levels. IgG and sIL-2R serum levels did not differ significantly between HPRL and NPRL patients: 1827.3 (1428-2226) vs 2028.8 (1586-2467) mg/dl and 882.2 (511-1254) vs 740.1 (534-946.4) U/ml, respectively (confidence interval 95%). In the total SLE group, sIL-2R and IgG serum levels were positively associated (P = 0.0009), however, this was not the case for sIL-2R and PRL (P > 0.49). We did not demonstrate an association between HPRL and lymphocyte activation measured through serum sIL-2R in female patients with SLE. PMID- 12630755 TI - High prevalence of antiphospholipid antibodies in Asian cancer patients with thrombosis. AB - Thrombotic complications are a significant cause of morbidity and mortality in cancer patients. Studies in Caucasian populations have shown that up to one-third of such patients test positive to antiphospholipid antibodies. Our aim was to determine the prevalence and serotypes of antiphospholipid antibodies in an unselected group of Asian cancer patients with thrombosis. All patients with cancer-related thrombosis seen in the Department of Hematology-Oncology and Radiation Oncology were enrolled in this study. The study period was from April 2000 to May 2001. Antiphospholipid antibodies tests were performed, namely lupus anticoagulant screen, anticardiolipin antibodies (IgG and IgM) and anti-beta-2 glycoprotein I antibodies (B2 GPI) IgG, IgM and IgA. Thirty-three patients were recruited. There were 14 males and 19 females, with an age range of 35-78 years of age. Of those enrolled, there were 25 Chinese, five Malays and three Indians. The patients had several cancer types: 11 (36.7%) patients had adenocarcinoma as the histological cell type. Of the 33 patients, 75.8% had stage IV disease. Arterial thrombosis was seen in eight patients (24.2%), and venous thrombosis occurred in 29 patients (87.9%). Antiphospholipid antibodies were positive in 60.6% of the patients, of which anti-B2GPI IgA antibody was the most prevalent antiphospholipid present (46.9%). The presence of anti-beta-2 glycoprotein I IgA antibody was associated with strokes, extensive and recurrent venous thrombosis, and coincident arterial and venous thrombosis. A high prevalence of antiphospholipid antibodies (60.6%) was found in Asian patients with cancer related thrombosis. The presence of antiphospholipid antibodies, particularly anti B2GPI IgA, may identify a subset of cancer patients who are at high risk of developing thrombotic complications, and further studies are warranted. PMID- 12630756 TI - Detection and clinical significance of Ba2+- and Sr2+-dependent antiprothrombin antibodies in patients with systemic lupus erythematosus and antiphospholipid syndrome. AB - We conducted this study to confirm whether or not Ca2+-dependent antiprothrombin antibody (aPT) in patients' sera with systemic lupus erythematosus and/or primary antiphospholipid syndrome would react to prothrombin/phosphatidylserine in the presence of Sr2+, Mn2+ or Ba2+, or divalent metal ions like Ca2+, utilizing ELISA, and to analyze the clinical significance of these metal-dependent aPT. We found the presence of Ba2+- and Sr2+-dependent IgG, IgM and IgA-aPT in up to 65% of patients negative for Ca2+-dependent IgG-, IgM and IgA-aPT. Maximally 69% of them were complicated by antiphospholipid syndrome-related symptoms. These data suggest that the measurement of Ba2+- and Sr2+- along with Ca2+-dependent aPT may become a clinically useful tool to correctly detect patients with antiphospholipid-related complications. However, further study is needed to clarify the clinicopathological differences among these aPTs in the future. PMID- 12630757 TI - Homozygous single nucleotide polymorphism of the complement C1QA gene is associated with decreased levels of C1q in patients with subacute cutaneous lupus erythematosus. AB - We report an association between a non-familial form of photosensitive Lupus specific skin disease, subacute cutaneous lupus erythematosus (SCLE), and a new single nucleotide polymorphism (SNP) in the C1QA gene. We also describe an association between this SNP and lower levels of serum C1q. This SNP consists of adenine replacing the third guanine in the codon for aminoacid residue Gly70 (position excludes the 22 amino acid leading peptide) that is located in the second exon of the C1QA gene. We have designated this SNP C1qA-Gly70GGA (the GenBank sequence at this location is C1qA-Gly70GGG). A survey of 19 SCLE patients showed that 11 (58%) were homozygous for C1qA-Gly70GGA SNP, seven (37%) were heterozygous, and only one patient (5%) was homozygous for the GenBank sequence. In contrast, only 13 of 62 (21%) normal controls were homozygous for the C1qA Gly70GGA SNP, 41 (66%) controls were heterozygous and eight (13%) controls were homozygous for the GenBank sequence. Thus, the C1qA-Gly70GGA SNP is strongly associated with SCLE (P-value = 0.005 by chi-square analysis with Yates correction). This SNP would traditionally be classified as clinically silent as it does not encode a different amino acid. However, our studies have suggested that this SNP appears to be associated with a functional abnormality of C1q expression since its presence correlates inversely with serum levels of C1q antigenic protein in both SCLE patients and normal controls. The mechanism by which this phenotypic change is associated with the translationally silent (synonymous) ClqA-Gly70GGA genetic variation is currently unknown. PMID- 12630758 TI - Treatment with sildenafil for the healing of refractory skin ulcerations in the antiphospholipid syndrome. AB - Nonhealing skin ulceration is a cutaneous manifestation of the antiphospholipid syndrome (APS) and is associated with thrombosis of small dermal vessels. Numerous therapies have been described but are either ineffective or require a prolonged course of treatment with potential complications. We describe the efficacy of sildenafil, a phosphodiesterase inhibitor, in the treatments of such ulcers. PMID- 12630760 TI - Catastrophic antiphospholipid syndrome complicating orthotopic liver transplantation. AB - Catastrophic antiphospholipid syndrome (CAPS) is an acutely devastating situation characterized by widespread thrombotic microangiopathy in the presence of elevated titers of antiphospholipid antibodies. We describe a 57-year old woman who underwent liver transplantation for primary sclerosing cholangitis and developed this malignant variant of the antiphospholipid syndrome. PMID- 12630759 TI - Pancreatitis leading to thrombotic thrombocytopenic purpura in systemic lupus erythematosus: a case report and review of literature. AB - Pancreatitis is a well-established but unusual complication of thrombotic thrombocytopenic purpura (TTP). It is also an unusual complication of systemic lupus erythematosus (SLE). However, TTP occurring as a consequence of acute pancreatitis in a patient with SLE has never been reported. We report a 24-year old African American woman with active systemic lupus (SLE) who developed thrombotic thrombocytopenic purpura (TTP) following an episode of acute pancreatitis. The TTP was manifested by low-grade fever, microangiopathic hemolytic anemia, renal insufficiency, altered mental status, seizures and thrombocytopenia. The patient was initially treated with pulse corticosteroids with inadequate response and subsequently with daily plasmaphresis, leading to full remission. This case represents first report of pancreatitis leading to TTP in a patient with systemic lupus erythematosus. PMID- 12630761 TI - Severe thrombocytopenic purpura due to rubella infection in a patient with systemic lupus erythematosus. AB - We report the case of a woman suffering from systemic lupus erythematosus who developed a severe thrombocytopenic purpura (platelet count < 1 x 10(9)/l) secondary to rubella infection. The search for antiplatelet antibodies revealed transient circulating anti-GPIIb-IIIa and anti-GPIb-IX platelet antibodies. After a few weeks, bound anti-GPIIb-IIIa antiplatelet antibodies were still detectable and they persisted several months after recovery, probably in relation to a mild autoimmune thrombocytopenia related to systemic lupus erythematosus. To our knowledge, this is the first case report of severe thrombocytopenic purpura due to rubella in an adult with systemic lupus erythematosus. PMID- 12630762 TI - Angiotensin-converting enzyme insertion/deletion polymorphism and vascular impairment in SLE. PMID- 12630763 TI - Tissue-polypeptide-specific antigen in SLE patients treated with low doses of quinagolide. PMID- 12630764 TI - The feasibility and safety of immunotherapy with dendritic cells loaded with CEA mRNA following neoadjuvant chemoradiotherapy and resection of pancreatic cancer. AB - BACKGROUND: Resected pancreatic cancer has a high risk of recurrence and mortality despite the the use of chemoradiotherapy. Because pancreatic cancers express tumor antigens such as carcinoembryonic antigen (CEA), it may be possible to immunize patients to induce tumor antigen-specific immune responses. We hypothesize that high-frequency tumor antigen-specific immune responses will reduce recurrence and increase survival. Autologous dendritic cells (DCs) loaded with tumor antigens are particularly potent at inducing tumor antigen-specific immune responses. METHODS: Three patients with resected pancreatic adenocarcinoma following neoadjuvant chemoradiotherapy received autologous, monocyte-derived DCs loaded with the mRNA encoding CEA monthly for 6 mo. RESULTS: It was feasible to generate an adequate number of DC from these patients and to cryopreserve them for repeated use. The DC demonstrated the typical immature phenotype. The immunizations were well-tolerated without evidence of adverse events. All three developed injection site reactivity. All three are alive without evidence of disease at more than 2 1/2 yr from the original diagnosis. CONCLUSION: The postoperative period following neoadjuvant chemoradiotherapy and pancreaticoduodenectomy for pancreatic cancer is an ideal environment to test novel immune-based therapies. DC-based immunotherapy in this setting is safe and feasible and may lead to prolonged survival. PMID- 12630767 TI - Bile duct brushings in a pig model: examination of intraobserver variability and variability in specimen quality obtained in sequential animals and between two different brushes. AB - PURPOSE: In patients with bile duct malignancy, bile duct brushing is plagued by a low yield diagnosing underlying malignancy. There are few data explaining why this is so. This porcine model was designed to examine three variables: 1) examination of inter-observer variability, 2) variability in specimen quality obtained in sequential animals, and 3) variability between two different brushes (one designed for colon with large bristles, one for duodenum with short bristles). METHODS: En bloc resection of liver, bile ducts, duodenum, and pancreas was performed on three 6-mo-old crossbred pigs at the time of commercial slaughter. In each pig, one common hepatic duct and one common bile duct brushing, all performed by the same investigator, were done. Ten identical vigorous passes were done with each brush (long bristle or short bristle) on virgin epithelium. Specimens were graded for cellularity by three cytopathologists who were blinded to the site or brush size. Interobserver variability as well as variability among sequential animals and between the two different brushes was compared. RESULTS: Interobserver variability among the three cytologists was almost nil. Cellularity obtained using the short brush alone varied from unsatisfactory to high. Cellularity obtained using the long brush alone varied from unsatisfactory to moderate. Variability of cells obtained from one pig to the next ranged from unsatisfactory to high. CONCLUSIONS: 1) While interobserver variability was very low, variability in cellularity obtained from one pig to the next, and from one brush to the next, was very high. This sampling variability may partially explain the low yield in malignant cells in human malignant biliary brushing. Multiple brushings in one patient may alleviate part of this problem. 2) There was no advantage to either brush type (large bristle or small bristle). PMID- 12630766 TI - The preventive effect of ketoconazole on experimental metastasis from a human pancreatic carcinoma may be related to its effect on prostaglandin synthesis. AB - BACKGROUND: Arachidonic acid metabolites known to affect platelet function also interfere with tumor growth and metastases. The purpose of this study was to evaluate the anti-metastatic potential of ketoconazole, a thromboxane synthetase and 5-lipoxygenase inhibitor, on hepatic metastasis from a human pancreatic adenocarcinoma in nude mice and its effect on serum prostaglandin levels. METHODS: The human pancreatic tumor cells (RWP-2) were injected intrasplenically in nude mice grouped into control, ketoconazole (270 microg), ketoconazole (360 microg), and ketoconazole (540 microg). The agent was administered intraperitoneally 30 min before and every 24 h after the tumor cell inoculation for 8 days. In a separate experiment thromboxane B2 (TxB2), prostaglandin D2 (PGD2), prostaglandin E2 (PGE2) and 6-Keto-F1a (stable prostacyclin derivative) were measured on blood from controls, tumor bearing animals and animals bearing tumors treated with 270 microg of ketoconazole. RESULTS: Statistically significant differences were observed between the control and three-treatment groups on the reduction of liver tumor nodules (p < 0.001), and in the liver surface areas occupied by tumor (p < 0.001). The TxB2 levels decreased from 150.6 ng/mL in the tumor bearing to 104.8 ng/mL in the ketoconazole treated animals (p < 0.05). PGD2, PGE2 and 6-keto-F1a levels increased to 7.1 ng/mL, 8.3 ng/mL, and 13.6 ng/mL from 3 ng/mL, 5.8 ng/mL, and 0.02 ng/mL respectively (p < 0.001). CONCLUSIONS: These results indicate that ketoconazole significantly reduced hepatic metastases from the human pancreatic carcinoma RWP-2 in the nude mouse model, and inhibited thromboxane B2 formation, potentiating a concomitant redirection of platelet endoperoxide metabolism into PGD2, PGE2, and 6-keto-F1a. It is hypothesized that the changes in the arachidonic acid metabolism mediate the ameliorating effect of ketoconazole on experimental hepatic metastasis. PMID- 12630765 TI - Peroxisome proliferator-activated receptor gamma ligand inhibits cell growth and invasion of human pancreatic cancer cells. AB - BACKGROUND: Peroxisome proliferator-activated receptor gamma (PPARgamma) is expressed in certain human cancers; ligand-induced PPARgamma activation can result in growth inhibition and differentiation in these cells. However, the precise mechanism for the antiproliferative effect of PPARgamma ligands is not entirely known. AIM OF STUDY: The purpose of this study was to examine the effect of PPARgamma ligands on pancreatic cancer cell growth and invasiveness. METHODS: The effect of two PPARgamma ligands, 15 deoxy-delta12,14 prostaglandin J2 (15d PGJ2) and ciglitazone, on the growth of four human pancreatic cancer cell lines (BxPC-3, MIA PaCa-2, Panc-1, and L3.6) was assessed. Expression of cell-cycle and apoptotic-related proteins was measured. Finally, the effect of 15d-PGJ2 on pancreatic cancer cell invasiveness and matrix metalloproteinase expression was determined. RESULTS: Both 15d-PGJ2 and ciglitazone inhibited the growth of all four pancreatic cancer cell lines in a dose- and time-dependent fashion. Treatment of BxPC-3 cells with 15d-PGJ2 resulted in a time-dependent decrease in cyclin D1 expression associated with a concomitant induction of p21waf1 and p27kip1. In addition, 15d-PGJ2 treatment induced apoptosis through activation of caspase-8, -9, and -3. Moreover, pancreatic cancer cell invasiveness was significantly suppressed after treatment with a nontoxic dose of 15d-PGJ2, which was associated with a reduction of MMP-2 and MMP-9 protein levels and activity. CONCLUSIONS: These results demonstrate that PPARgamma ligands have the dual advantage of inhibiting pancreatic cancer cell growth while reducing the invasiveness of the tumor cells, suggesting a potential role for these agents in the adjuvant treatment of pancreatic cancer. PMID- 12630768 TI - Pretreatment CA 19-9 level as a prognostic factor in patients with advanced pancreatic cancer treated with gemcitabine. AB - BACKGROUND: Serum levels of CA 19-9 correlate with survival among patients with pancreatic cancer treated with surgery or radiation therapy. In addition, CA 19-9 responses have been shown to predict for a better prognosis among patients with advanced disease treated with chemotherapy. The present study evaluates the predictive role of CA 19-9 pretreatment levels and response among patients treated with gemcitabine. METHODS: We retrospectively identified 28 patients with advanced pancreatic cancer and baseline elevations of CA 19-9 (> 37 U/mL) who were treated with single agent gemcitabine. CA 19-9 response was defined as a > or = 50% decline at any time after treatment. Survival was estimated with the Kaplan-Meier method, and curves were compared with the log-rank test. RESULTS: Eleven patients (39%) had a CA 19-9 response. The median survival of responding patients was longer than that of non-responding patients (13.8 vs 8 mo, p = .0272). When pretreatment CA 19-9 levels were analyzed, patients who had CA 19-9 below the median for the entire sample (1212 U/mL) lived significantly longer than patients with a CA 19-9 above the median (14.9 vs 7.4 mo, p = .0013). On multivariable analysis, pretreatment CA 19-9 level was an independent, and stronger predictor of survival (p = .0005) than CA 19-9 response (p = .0497). Other variables were not associated with survival. CONCLUSIONS: CA 19-9 may be a useful adjunct to response evaluation is this setting. In addition to CA 19-9 responses, prechemotherapy levels of this marker seem to have strong prognostic significance. PMID- 12630769 TI - Intraductal oncocytic papillary neoplasm of the pancreas shows strong positivity on FDG-PET. PMID- 12630770 TI - Solid-pseudopapillary tumor of the pancreatic head causing marked distal atrophy: a tumor originated posterior to the main pancreatic duct. AB - We report the case of a solid-pseudopapillary tumor (SPT) of the head of the pancreas causing occlusion of the main pancreatic duct (MPD) and marked pancreatic atrophy distal to the tumor disproportionate to the tumor size. A 15 yr-old girl was diagnosed with 5-cm solid-pseudopapillary tumor of the pancreatic head with marked distal pancreatic atrophy. Endoscopic retrograde cholangiopancreatography demonstrated obstruction of the MPD in the pancreatic head. We performed a duodenum-preserving pancreatic head resection to avoid postoperative exocrine and endocrine insufficiency. The surgical specimen showed the typical gross appearance of a SPT, with only a thin rim of pancreas anterior to the tumor. We believe that this presentation results when a tumor originates posterior to the MPD. Thus, whether or not pancreatic atrophy occurs depends strongly on the anterior/posterior relationship between the enlarging tumor and the MPD. The risk of SPT causing severe pancreatic atrophy should be kept in mind to avoid irreversible pancreatic insufficiency in young females. PMID- 12630773 TI - Modelling plant virus epidemics in a plantation-nursery system. AB - The material used for propagation and planting of many perennial crop plants is derived from vegetative cuttings which are first multiplied in a nursery. This situation was modelled to analyse the dynamics of a plant virus epidemic in a combined nursery-plantation system and the comparative effects of disease management activities in the plantation and in the nursery. The plant populations were partitioned into healthy and diseased categories and were linked according to basic SI models of disease transmission. Removal of diseased plants and replanting operations in both the plantation and nursery were included in the model and two variants were analysed in which mother plants (from which cuttings were taken) remained in the plantation or were harvested. The former is shown to be the limiting case for a large number of cuttings per plant. A criterion was derived for the invasion of disease into a healthy combined system. This consisted of four additive terms: the basic reproductive numbers of disease in the plantation alone and in the nursery alone, and two terms describing the cycling of diseased material between the plantation and nursery. Disease can still invade the system with basic reproductive numbers in the plantation or in the nursery less than 1 depending on the magnitude of cycling. Under some conditions only diseased plants remain in the plantation and nursery. For such a case a criterion was derived for the invasion of healthy plants into a fully diseased system. This depended on replanting rates in the plantation and nursery, and infection, mortality and removal rates of healthy plants. PMID- 12630772 TI - Barrett's esophagus and squamous cell carcinoma in a patient with psychogenic vomiting. AB - We report the association of Barrett's esophagus and invasive squamous cell carcinoma of the distal esophagus in a young 31-yr-old woman with a history of self-induced psychogenic vomiting. The development of intestinalized columnar mucosa and esophageal cancer in this young patient illustrates the complicated associations between human behavior and pathogenetic mechanisms involved in esophageal carcinogenesis. PMID- 12630771 TI - Sentinel lymph node metastasis in anal melanoma: a case report. AB - Anal melanoma represents only 1% of all melanomas. Owing to delayed diagnosis and early metastasis, the prognosis is uniformly poor. Sentinel lymph node (SLN) biopsy has become the preferred method of nodal staging method for cutaneous melanoma. The role of SLN biopsy for staging of anal melanoma remains unclear. We report a 39-yr-old Caucasian woman who presented with a history of chronic hemorrhoidal pain. She noted a pedunculated peri-anal mass associated with bleeding. Upon biopsy, the lesion was found to be a 6-mm thick primary anal melanoma. There was no evidence of metastatic disease on preoperative imaging studies. She underwent wide local excision of the peri-anal site of the primary melanoma and intra-operative lymphatic mapping with both isosulfan blue and filtered technetium sulfur colloid. With the guidance a lymphoscintigram, ipsilateral inguinal sentinel lymphadenectomy identified five nodes, all of which were both "hot" and blue. One node was found to have a 1-mm metastatic deposit. Subsequently, the patient was treated with adjuvant radiation therapy to the primary site as well as to the superficial and deep inguinal nodal basins. She also received four cycles of biochemotherapy. SLN biopsy appears feasible for staging the superficial inguinal lymph nodes in patients with anal melanoma. However, the impact of SLN biopsy, early detection of occult metastasis, and adjuvant systemic and radiation therapy on the long-term survival of patients with anal melanoma is uncertain. PMID- 12630774 TI - Population-induced oscillations in blended SI-SEI epidemiological models. AB - The effects of standard patterns of population growth in blended SI-SEI models (of which models for 'fast and slow' tuberculosis are an instance) are considered. When the incidence has the 'true mass action' form, the system is globally stable under both exponential and logistic population dynamics, whereas sustained oscillations occur in the case of bilinear incidence. This shows in the final analysis, the minimal dynamical ingredients needed to generate oscillations in basic epidemiological models: provided the population is exponentially growing and the incidence is bilinear, at least a fraction of the newly exposed individuals must enter the infective state with a delay. PMID- 12630775 TI - The effects of re-infection in directly transmitted infections modelled with vaccination. AB - We propose a mathematical model to deal with directly transmitted infections incorporating the loss of immunity. The model is developed taking into account a constant contact rate among individuals and an age-dependent vaccination rate. Based on this model, we analyse the effects of re-infection in a community under a vaccination strategy. PMID- 12630776 TI - A delay differential equation model on harmful algal blooms in the presence of toxic substances. AB - The periodic nature of blooms is the main characteristic in marine plankton ecology. Release of toxic substances by phytoplankton species or toxic phytoplankton reduce the growth of zooplankton by decreasing grazing pressure and have an important role in planktonic blooms. A simple mathematical model of phytoplankton-zooplankton systems with such characteristics is proposed and analysed. As the process of liberation of toxic substances by phytoplankton species is still not clear, we try to describe a suitable mechanism to explain the cyclic nature of bloom dynamics by using different forms of toxin liberation process. To substantiate our analytical findings numerical simulations are performed and these adequately resemble the results obtained in our field study. PMID- 12630777 TI - In situ SEM imaging at temperatures as high as 1450 degrees C. AB - Modifications to a scanning electron microscope (SEM) and a commercially available heating stage permits in situ imaging at temperatures as high as 1450 degrees C. Here we report on the technical modifications necessary to allow such high-temperature in situ imaging. In addition, in order to underline the potential of this technique in the field of materials science, three heating stage experiments are presented, which reveal microstructural changes occurring at high temperature. The respective in situ experiments are: (i) surface crystallization of a cordierite glass at 1050 degrees C; (ii) thermal recovery of asbestos (chrysotile) fibers at 1250 degrees C; and (iii) residual pore-structure evolution of tricalcium phosphate during sintering at 1450 degrees C. PMID- 12630778 TI - Imaging of a single atomic column in silicon grain boundary. AB - Atomic resolution high voltage electron microscopy was applied in investigations of the atomic structure of [112] sigma3 CSL grain boundary of silicon. Images of the grain boundary, viewed in the < 110 > direction, showed two types of dark spots. One type was rod shaped, which represented an atomic pair aligned in the < 111 > direction. The other was a small round spot, which represented a single atomic column in the grain boundary. The atomic structure of [112] sigma3 CSL grain boundary was directly shown from the atomic-structure image. PMID- 12630779 TI - Development of in situ observation technique using scanning ion microscopy and demonstration of Mn depletion effect on intragranular ferrite transformation in low-alloy steel. AB - An in situ observation technique using scanning ion microscopy (SIM) to investigate the behaviour of phase transformation from austenite to ferrite in low-alloy steel is proposed. The most important advantage of the technique is the combined observation by SIM and fabrication by focused ion beam (FIB) at a wide range of temperature, from room temperature to 1673 K. Using this advantage, we have developed a new technique for observing intragranular ferrite (IGF) transformation in low-alloy steel, which is the phase transformation occurring at an inclusion existing inside a grain of austenite phase. We successfully observed the IGF transformation at 873 K by examining an inclusion exposed to the specimen surface by FIB fabrication at 1073 K during cooling from 1673 K. From the results of the in situ observation, we determined the role of Mn depletion due to precipitation of MnS for IGF transformation in steel. PMID- 12630780 TI - Contrast-to-gradient method for the evaluation of image resolution in scanning electron microscopy. AB - In a previous study, we proposed the contrast-to-gradient (CG) method for evaluating image resolution. Here, the CG resolution is defined as a weighted harmonic mean of the local resolution, which is proportional to the quotient of the threshold contrast divided by the local gradient. The local gradient is calculated from the quadratic function that best fits the local pixel intensities over the region of interest (ROI) of 3 x 3 or 5 x 5 pixels in size. To refine the CG method, some modifications are carried out in the present study. Directional resolutions are employed to evaluate images, including astigmatism or strongly directional patterns as well as isotropic patterns. Here, CG resolution is redefined so as to keep the same value even for the image reversed in black-and white contrast, because of no difference in the image information during contrast reversing. Besides, CG resolution is characterized to be independent of the brightness/contrast change unless these changes do not bring about both cut-off and saturation in the pixel intensities. Dependencies of the denoising effect and the resolution accuracy on ROI size are demonstrated as a function of image noise. PMID- 12630781 TI - Retrieval process of high-resolution HAADF-STEM images. AB - A process for retrieving experimental high-resolution high-angle annular dark field (HAADF) scanning transmission electron microscopy (STEM) images, which are systematically distorted by environmental and/or instrumental instability in addition to the convolution influence of the probe, is demonstrated using a HAADF STEM image of an antimony-rich basal plane inversion boundary in Sb2O3-doped ZnO ceramics recorded in the [0110] zone axis. The process includes a correction of the diffractogram of the HAADF-STEM image by referring to the diffractogram of a high-resolution transmission electron microscopy image of the same area. The corrected diffractogram is used for deconvolution processing combined with the maximum entropy method. The retrieved images show bright spots regarded as the object function. Thus, the processing of the experimental HAADF-STEM image provides almost a real projected atomic structure by reforming the systematic distortion and eliminating the effects of the probe function. PMID- 12630782 TI - Mapping of sp2/sp3 in DLC thin film by signal processed ESI series energy-loss image. AB - A set of signal processing methods comprising fast Fourier transform interpolation, maximum entropy deconvolution and wavelet transformation has been successfully integrated to improve the equality of the extracted C K-edge spectra from electron spectroscopic imaging (ESI) series. Fast Fourier transform interpolation is used to improve the dispersion arising from discrete sampling of ESI series in the energy space. The maximum entropy method is used to dispel the convolution effect resulting from that ESI series acquired with a finite energy window. Wavelet transformation is applied to de-noise the extracted ESI spectrum. The post-processed ESI spectrum has quality as good as that of a probe-acquired spectrum and makes semi-quantitative analysis of the two-dimensional sp2/sp3 ratio map in diamond-like carbon thin film possible. In general, this method is applicable for reconstructing good quality core-loss electron energy-loss spectra from a nanometre-sized area, so that it may be possible to quantitatively analyse two-dimensional information about electronic structure in materials with near nanometre resolution. PMID- 12630783 TI - The specific heat of the cuticle and the morphological differences between the brown and yellow cuticles of hornets. AB - The morphology and thermophysical properties of stripes of brown and yellow cuticle collected from specimens of the Oriental hornet, Vespa orientalis (Hymenoptera, Vespinae), were analysed and compared by scanning and transmission electron microscopy. In the brown cuticle the colour stems from incrustation of the pigment melanin. In the yellow cuticle the yellow coloured pigment is concentrated in pockets between two layers of cuticle and its colour shows through the transparent cuticle. Both brown and yellow cuticles are composed of more than 30 layers. The upper layers are about 1-4 microm thick and as we proceed inwards, they become thinner so that the deepest ones that can still be discerned are thinner than the upper-most layer by one or two orders of magnitude (i.e., x 1/10 - x 1/100). The brown and yellow cuticles are each about 20 microm thick, but owing to the yellow pigment inserted in pockets of the yellow cuticle the total thickness of the latter is about 45 microm. The morphology of the yellow granules that contain the yellow pigment is shown and discussed. Below 20 degrees C and above 40 degrees C the specific heat (Cp) readings for both types of cuticle coincide, their values lying between approximately 1.55 and 1.60 J kg( 1), but in the temperature range 20-40 degrees C, the specific heat of the yellow strips displays a sharply non-monotonic behaviour, attaining a value of Cp = 1.8 J / Kg at 30 degrees, which contrasts with the relatively smooth curve shown by the brown strips. This anomaly (hump) does not occur following preheating of the samples to 70 degrees C. The shape of the repeated (annealed) 'yellow' curve is basically the same as that of the brown curve. The pockets of yellow pigment and their thermal contribution can be evaluated quantitatively by integration and correspond to an enthalpy of H = 3.5 J g(-1). The difference between the two types of hornet cuticle and the significance of this difference is discussed. PMID- 12630785 TI - Discontinuous capillary segments in the extensor digitorum longus muscle of aged BUF/Mna rats. AB - We examined the structural changes of capillaries in the extensor digitorum longus muscle of aged (25 months) male BUF/Mna rats, which caused severe muscle weakness of hind legs during aging. The aged muscle mostly consisted of bundles of muscle fibres 15-35 microm in diameter (type 1). In some small areas, however, muscle bundles contained small muscle fibres mainly 15-25 microm in diameter (type 2), possibly indicating that these small fibres are in the course of regeneration after necrosis. Examination of serial ultrathin sections revealed some remarkable changes of capillaries in the type 2 muscle bundle. In some capillaries, the vascular lumen became discontinuous by several close contacts of opposed endothelial cells in their course, forming plural closed vascular segments. Moreover, a solitary endothelial cell was often observed within a scaffold of basal lamina, which remained after destruction of endothelial cells. The segmentation of capillaries and the occurrence of the scaffolds of basal laminae are considered to indicate the degenerative process of capillaries. In some instances, on the other hand, endothelial cells closely apposed each other with no vascular lumen for distances of up to 100 microm, and some capillaries had a narrow vascular lumen (1-3 microm diameter) for a long distance (approximately 300 microm), probably indicating that these structures are in the course of regenerating capillaries to remodel the capillary networks around the muscle fibres. Pericytic processes circularly arranged outside the endothelium at the slit-like and narrow vascular lumen, like hoops, possibly preventing the rupture of the newly-formed vascular lumen from the increased blood flow and/or blood pressure. In addition, the occasional occurrence of capillaries with fenestrations may participate to increase the supply of nutrients and oxygen to the regenerating muscle fibres. The present findings suggest that the capillary networks in the extensor digitorum longus muscle of aged BUF/Mna rats are remodelled following the regeneration of muscle fibres after necrosis, and that on occasion, neighbouring endothelial cells may closely contact with each other both in the degenerative and regenerative processes of capillaries. PMID- 12630784 TI - Effects of enamel matrix derivative on mineralized tissue formation during bone wound healing in rat parietal bone defects. AB - Enamel matrix derivative (EMD: Emdogain) has been reported to stimulate the biosynthesis and regeneration of trabecular bone. To address whether the biological action of EMD is dependent on the local environment of osseous tissue, circular perforations were made in parietal bones and immediately filled with either EMD or its carrier, propylene glycol alginate (PGA), as control. On post operative days 4-60, the dissected bones were examined by various histological techniques. New bone matrix, which was immunoreactive for bone sialoprotein (BSP), was formed from the periosteum at the peripheral area of perforations. Different from the findings reported in injured long bones, mineralized tissue was produced in the regenerating connective tissue within bone defects. This mineralized tissue was hardly immunostained for BSP, contained few collagen fibres, and lacked osteocytic lacunae and layers of osteoblasts and osteoid. Energy-dispersive X-ray analysis showed that Ca and P weight % and Ca/P molar ratio of this mineralized tissue were similar to or slightly higher than those in the pre-existing parietal bones. In addition, most multinucleated cells located in mineralized tissue lacked a ruffled border structure and showed weak immunoreaction for the lysosomal cysteine proteinase, cathepsin K, whereas those located in the bone matrix exhibited ruffled borders and strong cathepsin K expression. However, multinucleated cells located in both tissues were strongly stained for tartrate-resistant acid phosphatase. The volume fraction of such mineralized tissue appeared to be higher in EMD-applied bones than in PGA-applied controls. The mineralized tissue-forming stromal cells within bone defects appeared to show greater accumulation in EMD-applied bones than in PGA-applied controls. Our results suggest that the bioactive effects of EMD on bone wound healing and mineralized tissue formation depend, at least in part, on the local osseous environment where EMD has been applied. PMID- 12630786 TI - Does size matter for sports performance researchers? PMID- 12630787 TI - Modelling the flight of a soccer ball in a direct free kick. AB - This study involved a theoretical and an experimental investigation of the direct free kick in soccer. Our aim was to develop a mathematical model of the ball's flight incorporating aerodynamic lift and drag forces to explore this important 'set-play'. Trajectories derived from the model have been compared with those obtained from detailed video analysis of experimental kicks. Representative values for the drag and lift coefficients have been obtained, together with the implied orientation of the ball's spin axis in flight. The drag coefficient varied from 0.25 to 0.30 and the lift coefficient from 0.23 to 0.29. These values, used with a simple model of a defensive wall, have enabled free kicks to be simulated under realistic conditions, typical of match-play. The results reveal how carefully attackers must engineer the dynamics of a successful kick. For a central free kick some 18.3 m (20 yards) from goal with a conventional wall, and initial speed of 25 m x s(-1), the ball's initial elevation must be constrained between 16.5 degrees and 17.5 degrees and the ball kicked with almost perfect sidespin. PMID- 12630788 TI - Dynamics of visual feedback in a laboratory simulation of a penalty kick. AB - Sport scientists have devoted relatively little attention to soccer penalty kicks, despite their decisive role in important competitions such as the World Cup. Two possible kicker strategies have been described: ignoring the goalkeeper action (open loop) or trying to react to the goalkeeper action (closed loop). We used a paradigm simulating a penalty kick in the laboratory to investigate the dynamics of the closed-loop strategy in these controlled conditions. The probability of correctly responding to the simulated goalkeeper motion as a function of time available followed a logistic curve. Kickers on average reached perfect performance only if the goalkeeper committed him or herself to one side about 400 ms before ball contact and showed chance performance if the goalkeeper motion occurred less than 150 ms before ball contact. Interestingly, coincidence judgement--another aspect of the laboratory responses--appeared to be affected for a much longer time (> 500 ms) than was needed to correctly determine laterality. The present study is meant as groundwork for experiments in more ecological conditions applicable to kickers and goalkeepers. PMID- 12630789 TI - Heart rate, blood lactate concentration and estimated energy expenditure in a semi-professional rugby league team during a match: a case study. AB - The aim of this study was to examine heart rate, blood lactate concentration and estimated energy expenditure during a competitive rugby league match. Seventeen well-trained rugby league players (age, 23.9 +/- 4.1 years; VO2max, 57.9 +/- 3.6 ml x kg(-1) x min(-1); height, 1.82 +/- 0.06 m; body mass, 90.2 +/- 9.6 kg; mean +/- s) participated in the study. Heart rate was recorded continuously throughout the match using Polar Vantage NV recordable heart rate monitors. Blood lactate samples (n = 102) were taken before the match, after the warm-up, at random stoppages in play, at half time and immediately after the match. Estimated energy expenditure during the match was calculated from the heart rate-VO2 relationship determined in laboratory tests. The mean team heart rate (n = 15) was not significantly different between halves (167 +/- 9 vs 165 +/- 11 beats x min(-1)). Mean match intensity was 81.1 +/- 5.8% VO2max. Mean match blood lactate concentration was 7.2 +/- 2.5 mmol x l(-1), with concentrations for the first half (8.4 +/- 1.8 mmol x l(-1)) being significantly higher than those for the second half (5.9 +/- 2.5 mmol x l(-1)) (P<0.05). Energy expenditure was approximately 7.9 MJ. These results demonstrate that semi-professional rugby league is a highly aerobic game with a considerable anaerobic component requiring high lactate tolerance. Training programmes should reflect these demands placed on players during competitive match-play. PMID- 12630790 TI - Differences between the sexes in the three-dimensional angular rotations of the lumbo-pelvic-hip complex during treadmill running. AB - The aims of this experiment were to determine whether there are differences between the sexes in the three-dimensional angular rotations of the lumbo-pelvic hip complex during running and, if such differences exist, to establish whether factors other than sex can explain the observed differences. A cohort of 44 non injured runners (22 males, 22 females) who usually ran more than 20 km per week were voluntarily recruited. All trials were conducted on a treadmill at a running speed of 4.0 m x s(-1). Reflective markers were placed over specific anatomical landmarks of the thoraco-lumbar spine, pelvis and femur. Data were captured using a VICON motion analysis system. Females tended to display a greater peak-to-peak oscillation for most of the angular rotations. An offset was the main difference between the male and female group mean waveforms for pelvis anterior-posterior tilt. Forward stepwise regression analysis revealed that sex was the most common variable related to the amplitudes of the angular rotations. Given these results, sport scientists conducting future biomechanical studies using angular data to test hypotheses are advised to be extremely cautious about averaging across male and female participants. PMID- 12630791 TI - Applying an extended version of the theory of planned behaviour to physical activity. AB - This prospective study explored whether extending the Theory of Planned Behaviour (TPB) with additional variables (descriptive norm, moral norm, anticipated affective reaction, self-identity and past behaviour) would account for additional variance in physical activity intentions and behaviour. Four interactions with past behaviour were also investigated. UK college employees participated in this study (n = 200, Time 1; n = 146, Time 2). Moral norm, self identity and past behaviour each explained additional variance in intentions, over and above the TPB variables. Past behaviour moderated the impact of descriptive norm on intentions. Intentions, self-identity and past behaviour were significant predictors of behaviour, as measured by a self-report physical activity questionnaire. To increase physical activity, interventions might target moral norm and self-identity for physical activity. Focusing on positive descriptive norms might benefit sedentary individuals. The significant role of past behaviour is less useful in directing interventions. PMID- 12630792 TI - Measurement error in short-term power testing in young people. AB - The aim of this study was to examine the consistency or reproducibility of measuring cycling peak power in children and adults. Twenty-seven pre-pubertal girls and boys and 27 female and male physical education students (age 9.8 +/- 0.5 and 24.4 +/- 4.3 years, respectively; mean +/- s) participated in the study. All participants performed five tests over 15 days and underwent a habituation session before the study. Each test included four sprints against four different braking forces. We found that braking forces of 7.5% of body weight in children and 10% of body weight in adults were too high for most of the participants to elicit maximal cycling power. Unlike the children, the physical education students improved their performance between session 1 and session 2 (1025 +/- 219 vs 1069 +/- 243 W; P < 0.001). Therefore, to obtain reproducible measures of cycling peak power, a habituation session including a complete test protocol (i.e. warm-up plus three sprints) is highly recommended. When the protocol included three sprints in children and at least two sprints in adults, measurement of cycling peak power was found to be highly reliable (test-retest coefficient of variation approximately 3%). Finally, to avoid performance fluctuations, especially over several consecutive evaluations (e.g. longitudinal studies), it is necessary to maintain high motivation in children. PMID- 12630793 TI - Making the case for personal professional monitoring in health care. PMID- 12630794 TI - Statistical process control tools for monitoring clinical performance. PMID- 12630795 TI - Risk-adjusted sequential probability ratio tests and longitudinal surveillance methods. AB - Regardless of the exact method employed, the application of statistical process controL SPRTs, or related longitudinal analysis methods can significantly improve the ability to monitor clinical processes and outcomes. Incorporation and adaptation of risk-adjustment and rare events into these methods represent important contributions to their use in health care. Fostering greater and more widespread use of these methods, however, remains a significant challenge. Hopefully studies such as those by Spiegelhalter et al. will lead to more awareness of their value for contributing to a safer health care system. PMID- 12630796 TI - Risk-adjusted sequential probability ratio tests: applications to Bristol, Shipman and adult cardiac surgery. AB - OBJECTIVE: To investigate the use of the risk-adjusted sequential probability ratio test in monitoring the cumulative occurrence of adverse clinical outcomes. DESIGN: Retrospective analysis of three longitudinal datasets. SUBJECTS: Patients aged 65 years and over under the care of Harold Shipman between 1979 and 1997, patients under 1 year of age undergoing paediatric heart surgery in Bristol Royal Infirmary between 1984 and 1995, adult patients receiving cardiac surgery from a team of cardiac surgeons in London,UK. MAIN OUTCOME MEASURE: Annual and 30-day mortality rates. RESULTS: Using reasonable boundaries, the procedure could have indicated an 'alarm' in Bristol after publication of the 1991 Cardiac Surgical Register, and in 1985 or 1997 for Harold Shipman depending on the data source and the comparator. The cardiac surgeons showed no significant deviation from expected performance. CONCLUSIONS: The risk-adjusted sequential probability test is simple to implement, can be applied in a variety of contexts, and might have been useful to detect specific instances of past divergent performance. The use of this and related techniques deserves further attention in the context of prospectively monitoring adverse clinical outcomes. PMID- 12630797 TI - Development of appropriateness criteria for colonoscopy: comparison between a standardized expert panel and an evidence-based medicine approach. AB - OBJECTIVES: To assess the degree of agreement between appropriateness criteria for the use of colonoscopy developed by a standardized expert panel method and evidence from published studies. DESIGN: Descriptive, agreement study. SETTING: Multidisciplinary panel; primary care practice in Switzerland. PARTICIPANTS: Nine national experts; 577 primary care patients referred for colonoscopy, 154 published papers. INTERVENTIONS: Evaluation of the appropriateness of 402 possible clinical indications for colonoscopy, based on a comprehensive review of the literature. MAIN OUTCOMES MEASURES: Proportion of agreement (weighted kappa), between panel- and literature-based appropriateness categories (appropriate, uncertain, inappropriate) for theoretical and actual indications encountered. RESULTS: Nineteen of 402 indications rated by the panel could be based on the evidence retrieved from eight randomized clinical trials. A 68% agreement (kappa = 0.52) was found between panel- and study-based criteria. The addition of an uncontrolled trial and seven observational studies yielded a 71% agreement (kappa = 0.63). Agreement was similar when examining 577 actual cases: 69% agreement, kappa = 0.47. Agreement between panel-based indications and published evidence was not influenced by the perceived comprehensiveness and the apparent quality of the published reports. CONCLUSIONS: Evidence for the appropriateness of most indications for colonoscopy could not be derived directly from the published literature. Agreement between appropriateness criteria developed by an expert panel and evidence from published studies was moderate to good, where available. New approaches should be sought in order to systematically integrate complementary evidence obtained from clinical trials and expert panels into practice guidelines. PMID- 12630798 TI - Why are people dissatisfied with medical care services in Lithuania? A qualitative study using responses to open-ended questions. AB - OBJECTIVES: To identify and describe the main sources of dissatisfaction with medical services among the population in Lithuania. DESIGN: Analysis of written responses to an open-ended question as a part of a questionnaire survey. PARTICIPANTS: A randomly selected group of 25--64-year-old men and women from the Utena and the Kaisiadorys regions of Lithuania. Of the 1395 survey respondents, 357 persons (25.6%) answered the question regarding the dissatisfaction with health care services. MAIN OUTCOME MEASURES: Identification of the sources of dissatisfaction with medical care services among study participants. RESULTS: Twelve categories of dissatisfaction were identified that were related to three levels: shortcomings in the health care system (systemic level), deficiencies in provision and quality of services (institutional level) and deficiencies in physicians' attitudes, skills and work (individual level). CONCLUSION: Consumers' perceptions of medical care quality can be analysed by using information obtained from simple survey material and can be used to improve the quality of service. The causes of dissatisfaction with medical service can be traced to the development of the health care delivery system and patients' rights in a country. PMID- 12630799 TI - Towards evidence-based clinical practice: an international survey of 18 clinical guideline programs. AB - OBJECTIVE: To describe systematically the structures and working methods of guideline programs. DESIGN: Descriptive survey using a questionnaire with 32 items based on a framework derived from the literature. Answers were tabulated and checked by participants. STUDY PARTICIPANTS: Key informants of 18 prominent guideline organizations in the United States, Canada, Australia, New Zealand, and nine European countries. MAIN OUTCOME MEASURES: History, aims, methodology, products and deliveries, implementation, evaluation, procedure for updating guidelines, and future plans. RESULTS: Most guideline programs were established to improve the quality and effectiveness of health care. Most use electronic databases to collect evidence and systematic reviews to analyze the evidence. Consensus procedures are used when evidence is lacking. All guidelines are reviewed before publication. Authorization is commonly used to endorse guidelines. All guidelines are furnished with tools for application and the Internet is widely used for dissemination. Implementation strategies vary among different organizations, with larger organizations leaving this to local organizations. Almost all have a quality assurance system for their programs. Half of the programs do not have formal update procedures. CONCLUSIONS: Principles of evidence-based medicine dominate current guideline programs. Recent programs are benefiting from the methodology created by long-standing programs. Differences are found in the emphasis on dissemination and implementation, probably due to differences in health care systems and political and cultural factors. International collaboration should be encouraged to improve guideline methodology and to globalize the collection and analysis of evidence needed for guideline development. PMID- 12630800 TI - Evaluating implementation of quality management systems in a teaching hospital's clinical departments. AB - OBJECTIVES: This study evaluated a strategy for implementing continuous quality improvement based on a decentralized quality management system in the clinical departments of a hospital. SETTING: The institution is a 2000-bed teaching hospital of tertiary health care employing 8000 people. METHODS: The quality management intervention was tested in six volunteer departments. This intervention comprised an instructional seminar, methodological assistance, and the dissemination of guidelines. The program was evaluated 1 year after the intervention and included a quality audit, interviews with department staff, and analysis of the written documents produced by the departments. RESULTS: The quality management systems are functioning in all the departments. Quality teams meet regularly and multidisciplinary work groups are in place. The topics most often addressed are patient reception and communication between department staff members. The level of compliance with the guidelines has increased, from 39% before the seminar to 54% 1 year later (P < 0.05). All of the staff members interviewed judged the process useful for them and for the department, while waiting for the concrete results. Among the difficulties the staff members encountered were changing their work habits, lack of time, and the tedious aspect of writing procedures. CONCLUSION: Implementing continuous quality improvement in hospital departments seems to be an interesting alternative to organization-wide implementation strategies. However, these results need to be confirmed by long term evaluations and by deploying the program i n other departments. PMID- 12630801 TI - Predictors of inappropriate hospital stay: a clinical case study. AB - OBJECTIVE: To assess the reasons for inappropriate patient stay (IPS) and to identify possible predictors of this inappropriate stay. DESIGN: The reasons for IPS were analyzed in a cross-sectional survey at two surgical one gynecologic and one obstetric ward, using a sample of 610 days of hospital stay by means of the Dutch Appropriateness Evaluation Protocol. SETTING: The study was conducted at the University Hospital Maastricht, a 715-bed hospital with a regional and teaching function, located in the southern part of The Netherlands. RESULTS: Results indicate that > 20% of the hospital stay was inappropriate. The reasons for IPS were statistically significantly related to the patients' age, the availability of home care and medical specialty. In a predictive model, only the specialty proved to be a predictor of IPS. Most of the IPS occurred during the first days of hospital stay and the days before the patient's discharge. CONCLUSION: A substantial proportion of hospital stay was found to be inappropriate, due to hospital procedures and the inability to refer patients to other care facilities or care providers. Analysis of the causes of IPS provided useful data for improvement actions. Efficient use of hospital resources should be promoted by reducing the delay in interventions and discharge procedures. PMID- 12630802 TI - Perceptions of health care quality in Central America. AB - OBJECTIVES: To examine user perceptions of health care delivery in selected rural and urban areas of three Central American countries. DESIGN: Three focus group studies were conducted in 1997-98 in Honduras, Costa Rica, and Panama. In each selected region, 10 to 15 groups met to discuss health services available, access to and use of the services, satisfaction with different aspects of care and suggestions for improvement. SETTING: Regions chosen represented the poorest areas in each country and the dominant health care systems in Central America: the Ministry of Health system and the Social Security system based on mandatory contributions. PARTICIPANTS: 351 residents from rural and urban communities represented different genders, ages, occupations, health, and socio-economic status. RESULTS: Participants considered private care to be the best, but too costly. Their main preoccupations focused on prompt access to trusted physicians, effective and inexpensive medication, and quality attention in public hospitals. Hondurans favor the personal care offered in public clinics and rural hospitals, and hope for improved medical services. In Costa Rica and Panama, users prefer Social Security clinics for the medical specialties and perceived sophisticated technology, despite delays and poor attention. The rural poor, especially indigenous people, voice basic needs with little regard for quality. CONCLUSIONS: Health care quality is extremely variable in the three regions, requiring increased community participation to improve. Focus groups offered important, confidential and cost-effective information on quality and breadth of health care delivery and should be part of quality monitoring initiatives. PMID- 12630803 TI - Quality of psychiatric care: validation of an instrument for measuring inpatient opinion. AB - OBJECTIVES: To validate a brief self-completed questionnaire for routinely assessing patients' opinions on the quality of care in inpatient psychiatric wards (Rome Opinion Questionnaire for Psychiatric Wards). DESIGN: A preliminary version was assessed for face and content validity by eight psychiatrists and two patient focus groups. The final version was evaluated for acceptability, factor structure, internal consistency, and test-retest reliability. SETTING: An inpatient psychiatric ward in a general hospital in Rome. STUDY PARTICIPANTS: The questionnaire was administered to all consecutive inpatients admitted over a 6 month period (n = 169). Test-retest reliability was evaluated by administering the questionnaire for a second time to 27 inpatients. MAIN OUTCOME MEASURES: Face and content validity: psychiatrists and focus groups' opinions on relevance, importance, and clarity, acceptability: inpatients' opinions on user friendliness; factor analysis: principal component analysis; internal consistency: Cronbach's alpha; test-retest reliability: Cohen's weighted kappa coefficient, intraclass correlation coefficient RESULTS: After evaluating face and content validity, the questionnaire was reduced to 10 items. Inpatients found the questionnaire to be acceptable. Factor analysis revealed that three factors professional qualities of staff, information received, and physical environment explained 67.2% of total variance. Cronbach's alpha was 0.82 for the questionnaire and 0.61, 0.71, and 0.35 for the three factors, respectively. Test retest reliability was good; weighted kappa higher than 0.9 for three items and 0.6-0.9 for seven items. The intraclass correlation coefficient was 0.80. CONCLUSIONS: The questionnaire seems to be adequate for evaluating patients' opinions on care in inpatient psychiatric wards. Because of its user friendliness, it may be particularly suitable for routine use. PMID- 12630804 TI - Critical appraisal of clinical performance measures in Germany. AB - BACKGROUND: One of the tasks of the Federal Co-ordinating Committee, as part of the self-governing health care system in Germany, is to develop clinical performance measures (CPMs). As these measures generally exert a strong impact on health care delivery, their methodological quality should meet the highest standards. OBJECTIVE: To develop, for use in the German health care system, methodological quality requirements for CPMs and performance assessment programs. MATERIALS AND METHODS: We assembled and reviewed internationally available materials on CPMs. From these we prepared lists of requirements for appraising the methodological quality of CPMs and CPM programs. PRODUCTS: Lists of criteria for use in appraising the appropriateness of the development, content, format and application of CPMs and their value to the health care system for performance measurement; a checklist for assessing the methodological quality of CPMs/CPM programs. CONCLUSION: The methods applied and the products from this project offer a useful framework for prospective CPM/CPM program developers and set a high threshold for the development of methodologically rigorous clinical performance measurement. PMID- 12630805 TI - Adrenomedullin selectivity of calcitonin-like receptor/receptor activity modifying proteins. AB - Co-expression of an initially orphan calcitonin receptor-like (CL)1 receptor with individual receptor-activity-modifying proteins (RAMP)1, -2 and -3 results in CL receptor/RAMP1, -2 and -3 proteins at the cell surface. The RAMP define the selectivity of the CL receptor for the vasodilatory peptides adrenomedullin (AM) and calcitonin gene-related peptide (CGRP). The selectivity for AM and CGRP agonists and antagonists of human, rat, porcine and bovine CL receptors, co expressed with RAMP2 and -3, has been studied in different cell types. This revealed CL receptor/RAMP2 and CL receptor/RAMP3 as AM1 and AM2 receptor subtypes, respectively. The AM1 receptor crossreacts with CGRP at high and the AM2 receptor at lower concentrations. Here the pharmacological properties of the cloned AM receptors are compared to those revealed in tissues and cell lines. According to nomenclature recommendation of the IUPHAR (International Union of Pharmacology) subcommittee XXXII, the former CRLR is now the CL receptor (1). PMID- 12630806 TI - Regulation of cell growth and apoptosis by adrenomedullin. AB - Adrenomedullin, originally discovered in human pheochromocytoma, has been shown to have potent vasodilatory activity. However, like other vasoactive peptide hormones, its physiological roles have been found to extend far beyond the regulation of vascular tonus, and to include such functions as the regulation of cell proliferation and differentiation. There is a growing body of evidence that adrenomedullin exerts a wide range of effects on cell growth and apoptotic death, and that these effects are dependent on cell type and experimental conditions. Signaling pathways independent of cyclic AMP, such as protein tyrosine kinase(s) and mitogen-activated protein kinases, may play key roles in the regulation of mitogenesis and apoptosis by adrenomedullin. PMID- 12630807 TI - Specificity of porcine calcitonin receptor and calcitonin receptor-like receptor in the presence of receptor-activity-modifying proteins. AB - Adrenomedullin (AM), calcitonin gene-related peptide (CGRP), amylin (AMY) and calcitonin (CT) are members of the CGRP/CT superfamily of peptides. Among them, AM and CGRP are reported to share a core receptor, the calcitonin receptor-like receptor (CRLR), and the specificity of the CRLR is determined by the expression levels of receptor-activity-modifying proteins (RAMPs). In the case of AMY, co expression of the calcitonin receptor (CTR) and RAMPs was recently reported to form its specific receptor. However, detailed analysis of the receptor specificity of the CRLR and CTR in the presence of RAMPs has so far been performed mainly in the human system. Thus, we cloned cDNAs encoding porcine CRLR, RAMP1, RAMP2 and RAMP3 precursors from a porcine lung and hypothalamus cDNA library, and determined their sequences. Then, porcine RAMPs, CRLR and CTR were expressed in COS-7 or porcine vascular smooth muscle cells, and the resulting receptor complexes were analyzed by the cyclic adenosine 3,5-monophosphate (cAMP) production assay. The specificity of CRLR was clearly determined by the expression of RAMPs; RAMP1 converted CRLR to CGRP receptor, while RAMP2 and RAMP3 converted it to AM receptor, but the affinity of CTR for AMY was not increased by the expression of any known RAMPs. In contrast to previous findings, porcine CTR and RAMP did not appear to form an AMY receptor having sufficient affinity and specificity for the physiological interaction. PMID- 12630808 TI - The function of extracellular cysteines in the human adrenomedullin receptor. AB - When co-expressed with receptor activity-modifying protein (RAMP) 2, calcitonin receptor-like receptor (CRLR) functions as an adrenomedullin (AM) receptor (CRLR/RAMP2). In the present study, we examined the function of the cysteine (C) residues in the extracellular loops of human (h)CRLR (C212, C225 and C282) and in the extracellular domain of hRAMP2 (C68, C84, C99 and C131). Using site-directed mutagenesis, the cysteine residues were substituted, one at a time, with alanine (A). Co-expression in HEK293 cells of hRAMP2 with the hCRLR C212A or C282A mutant significantly reduced the 50% of effective concentration (EC50) for AM-evoked cyclic adenosine monophosphate (cAMP) production, despite full cell surface expression of the receptor heterodimer. Co-expression of the C225A mutant had no effect on [125I]AM binding or receptor signaling. These results suggest that the cysteine residues in the first (C212) and the second (C282) extracellular loops form a disulfide bond that is important for stabilizing the receptor in the correct conformation for ligand binding and activation. Cells expressing hCRLR with an hRAMP2 mutant (C68A, C84A, C99A or C131A) showed no specific AM binding or AM-stimulated cAMP accumulation. Though abundant in the intracellular compartment, these receptors were not detected at the cell surface, suggesting that all four cysteine residues are essential for efficient transport to the plasma membrane. Cysteine residues in the extracellular loops of hCRLR and in the extracellular domain of hRAMP2 thus appear to play distinct roles in the cell surface expression and function of the receptor heterodimer. PMID- 12630809 TI - Immunohistological localization and possible functions of adrenomedullin. AB - In this short review, we describe the distribution of adrenomedullin (AM) immunoreactive cells in human tissues and their related biological properties, focusing on the blood coagulation and mucosal defense systems. AM is widely distributed in human tissues, especially in cardiovascular and endocrine tissues. Within vessels, AM has been immunohistochemically detected in vascular smooth muscle cells (SMCs) and endothelial cells (ECs). In atherosclerotic lesions, the peptide is present not only in these cells, but also in macrophages, and the most intense AM immunoreactivity is detected in macrophages located in shoulder lesions of atheromatous plaque, which are considered to be rupture-prone regions. AM inhibits tissue factor production, and augments the production and release of tissue factor pathway inhibitor from aortic ECs. AM also induces the release of antithrombin and urokinase-type plasminogen activator from ECs. Taken together, these antithrombotic properties of the peptide are expected to play an important role in the maintenance of blood circulation. Furthermore, AM immunoreactivity is observed in mucosal and glandular epithelia of the gastrointestinal, respiratory and reproductive systems. AM and the proadrenomedullin N-terminal 20 peptide (PAMP) show strong antibacterial activity against Escherichia coli. In addition, AM is also present in the auditory system. These lines of evidence suggest that AM and its related peptides not only play a role in vasodilatation, but also exhibit multiple biological activities in mammals. PMID- 12630810 TI - Decreased expression of adrenomedullin during adipocyte-differentiation of 3T3-L1 cells. AB - Adrenomedullin (AM) is a potent vasodilator peptide which has an inhibitory action on insulin secretion. Resistin is a novel peptide specifically secreted from adipocytes, and implicated in insulin resistance. We studied the expression of AM and resistin in 3T3-L1 adipocytes and preadipocytes by Northern blot analysis and radioimmunoassay. Immunoreactive-AM was detected in the culture media of 3T3-L1 preadipocytes and adipocytes, with higher concentrations found in preadipocytes. Northern blot analysis showed that AM mRNA was expressed in 3T3-L1 preadipocytes but was undetectable in adipocytes. In contrast, resistin mRNA was expressed in 3T3-L1 adipocytes, whereas it was not detected in 3T3-L1 preadipocytes. The present study thus showed that AM expression was decreased, and resistin expression increased, during adipocyte-differentiation of 3T3-L1 cells. PMID- 12630811 TI - Direct measurement of glycine-extended adrenomedullin in plasma and tissue using an ultrasensitive immune complex transfer enzyme immunoassay in rats. AB - The mature form of the vasodilator peptide adrenomedullin (AM-m) is synthesized from a glycine-extended precursor (AM-Gly) by enzymatic amidation. We have developed a highly sensitive enzyme immunoassay (Immune Complex Transfer Enzyme Immunoassay; ICTEIA) that enables us to measure levels of AM-Gly in plasma and tissue directly. The detection limit of this assay is 1 amol/assay, and the intra and inter-assay precision are 4.5-14.1% and 9.9-20.5%, respectively. Dilution curves for plasma samples showed good linearity, and the analytical recovery was 107-116.6%. Using ICTEIA, we determined that the plasma concentration of immunoreactive AM-Gly is substantially higher than that of AM-m (5.22 +/- 2.56 vs. 1.21 +/- 0.79 fmol/ml). In contrast, levels of AM-Gly were much lower than those of AM-m in the lung, heart, kidney, adrenal gland and liver. We also evaluated AM-Gly and AM-m levels in rats in a morbid state induced by intraperitoneal administration of lipopolysaccharide (LPS). In most tissues, levels of AM-m and AM-Gly were both increased by LPS; however, AM-Gly/AM-m ratios were not significantly affected, which suggests that AM-Gly is rapidly converted to AM-m in tissue. PMID- 12630812 TI - Mapping of the adrenomedullin-binding domains in human complement factor H. AB - Adrenomedullin (AM) is a multifunctional peptide involved in roles as varied as blood pressure regulation, growth, neurotransmission, and inflammation control, among others. We previously identified complement factor H as a serum binding protein for AM and showed that factor H regulates AM functions and vice versa. Here we searched for the specific binding sites for AM by using recombinant fragments of factor H and a non-radioactive binding assay with fluorescein-tagged AM. By this methodology, two specific binding sites for AM were found in factor H. One of them shows a high affinity for AM and is located at the carboxy terminal end of factor H, comprising short consensus repeats (SCR) 15-20. Smaller fragments of this region did not bind to AM efficiently, suggesting that the high affinity binding site of factor H requires a complex three-dimensional structure to recognize AM. Another binding site with lower affinity for AM was found in the middle of the factor H molecule, at SCR 8-11. Antibodies against factor H prevented AM binding altogether, but the main binding partner of factor H, C3b, did not, indicating that C3b and AM bind to different regions of factor H. These structure-function data support previous biochemical observations. Our understanding of the binding between AM and factor H may help in the development of new treatments for diseases in which these molecules play active roles. PMID- 12630813 TI - Adrenomedullin, an autocrine mediator of blood-brain barrier function. AB - Since the discovery that adrenomedullin gene expression is 20- to 40-fold higher in endothelial cells than even in the adrenal medulla, this peptide has been regarded as an important secretory product of the vascular endothelium, together with nitric oxide, eicosanoids, endothelin-1, and other vasoactive metabolites. Cerebral endothelial cells secrete an exceptionally large amount of adrenomedullin, and the adrenomedullin concentration is about 50% higher in the cerebral circulation than in the peripheral vasculature. The adrenomedullin production of cerebral endothelial cells is induced by astrocyte-derived factors. Adrenomedullin causes vasodilation in the cerebral circulation, may participate in the maintenance of the resting cerebral blood flow, and may be protective against ischemic brain injury. Recent data from our laboratory indicate that adrenomedullin, as an endothelium-derived autocrine/paracrine hormone, plays an important role in the regulation of specific blood-brain barrier properties. Adrenomedullin is suggested to be one of the physiological links between astrocyte-derived factors, cyclic adenosine 3'5'-monophosphate (cAMP), and the induction and maintenance of the blood-brain barrier. Moreover, the role of adrenomedullin in the differentiation and proliferation of endothelial cells and in angiogenesis suggests a more complex function for adrenomedullin in the cerebral circulation and in the development of the blood-brain barrier. PMID- 12630814 TI - Pathophysiological function of adrenomedullin and proadrenomedullin N-terminal peptides in adrenal chromaffin cells. AB - Adrenomedullin (AM) and peptides of the proadrenomedullin N-terminal 20 peptide (PAMP20) family are multifunctional peptides abundantly expressed in the adrenal medulla. These peptides are released by regulated exocytosis along with catecholamines upon stimulation of adrenal chromaffin cells. They are also released gradually during culture, and this release is stimulated by a 3',5' cyclic adenosine monophosphate (cAMP)-dependent pathway. The expression and release of AM increase under hypoxia in chromaffin cells. The expression of AM in pheochromocytoma PC12 cells is reduced during neuronal differentiation with nerve growth factor. On the other hand, PAMP20 and PAMP12 suppress catecholamine release and synthesis by interfering with nicotinic cholinergic receptors. AM increases blood flow in the adrenal gland, and causes a gradual release of catecholamine, but does not modify regulated exocytosis upon the stimulation of cells. Current data indicate that the expression of these peptides is regulated by intracellular signaling pathways, and changes under various physiological and pathological conditions. AM and PAMP20 family peptides have distinct physiological functions. PAMP20 and PAMP12 are endogenous peptides that modulate chromaffin cell function in an autocrine manner, whereas AM may mainly regulate vascular cell function in a paracrine manner. PMID- 12630816 TI - Role of the endogenous adrenomedullin system in regulating the secretion and growth of rat adrenal cortex. AB - The expression of components of the adrenomedullin (AM) system (AM and its receptors) has been detected in mammalian adrenal zona glomerulosa (ZG) cells, and evidence has been provided that AM is able to inhibit agonist-stimulated aldosterone secretion from and to enhance the proliferative activity of ZG cells. However, there has been no evidence that the endogenous AM system acts as a physiological regulator of ZG function. Hence, we investigated whether the suppression of AM gene transcription by a specific antisense oligodeoxynucleotide (ODN) is able to alter the secretion and growth of rat ZG cells cultured in vitro. ZG cell cultures were examined 0, 2, 4, 6 and 8 days after treatment with scrambled sense (S)-ODN (control cultures) and AM antisense (A)-ODN. Control cultures, as well as freshly dispersed ZG cells and ODN-untreated cultures, expressed AM as mRNA and protein. A-ODN treatment suppressed AM expression within 4 days and the suppression lasted until day 6. Confluent control cultures displayed basal and angiotensin-II (Ang-II), K(+)- and adrenocorticotropic hormone (ACTH)-stimulated aldosterone secretions similar to those of ODN untreated cultures. A-ODN treatment magnified the aldosterone response to Ang-II and K+ at days 4 and 6 (but not at day 8), without affecting the basal or ACTH stimulated secretion. As compared to ODN-untreated and control cultures, non confluent A-ODN-treated ones showed a 40% elongation in the duplication time, a significant decrease in the proliferation index, and a marked rise in apoptotic index from day 4 to day 8. In conclusion, our study validates the use of A-ODN to block the endogenous AM system, showing that suppression of AM-synthesis requires at least 2 days to become appreciable and persists for at least 6 days. Moreover, it provides the first evidence that endogenous AM plays a physiological role in cultured rat ZG cells, by exerting a buffering action on their acute secretory response to Ang-II and K+ and by maintaining normal basal proliferative and apoptotic activities. PMID- 12630815 TI - Endothelial responses of the aorta from adrenomedullin transgenic mice and knockout mice. AB - Adrenomedullin (AM) is a potent vascular wall-derived vasorelaxing peptide which induces the release of nitric oxide (NO). To explore the role of endogenous AM in vascular function, we examined the effects of acetylcholine (ACh), AM, and AM receptor antagonists [AM (22-52), and calcitonin gene-related peptide (CGRP) (8 37)] on the isometric tension of aortic rings isolated from AM transgenic (TG) and knockout (KO) mice and wild type littermates (WT). ACh and AM caused a dose dependent reduction of the isometric tension of aortic rings, but the degree of vasodilatation was smaller in TG than in KO or WT (% delta tension [10(-6) mol/l ACh]: KO -69 +/- 10%, WT -39 +/- 8%, TG -29 +/- 1%, p < 0.01). On the other hand, N(G)-nitro-L-arginine methyl ester, an NO synthase inhibitor, induced greater vasoconstriction in TG (% delta tension 10(-5)mol/l: KO +78 +/- 16%, WT +99 +/- 27%, TG +184 +/- 20%, p < 0.01), whereas E-4021, a cyclic guanosine monophosphate (cGMP)-specific phosphodiesterase inhibitor, caused greater vasodilation in TG mice. Both AM antagonists increased tension in TG to a greater extent than in KO or WT mice (% delta tension [10(-6) mol/l CGRP (8-37)]: KO +24 +/- 5%, WT +51 +/- 6%, TG +75 +/- 7%, p < 0.01). Endothelial denudation of the aorta diminished the vasoconstriction caused by the AM antagonists. In conclusion, the amounts of AM expressed in the aortic endothelium influenced baseline NO release. AM antagonists increased vascular tone in WT as well as in TG, suggesting that endogenous AM plays a physiological role in the regulation of aortic tone. PMID- 12630817 TI - Adrenomedullin promotes proliferation and migration of cultured endothelial cells. AB - Adrenomedullin (AM) is a vasoactive hormone which exerts its action through cyclic adenosine monophosphate(cAMP) /cAMP-dependent protein kinase (PKA) cascade and intracellular Ca2+ mobilization. Recently, evidence has accumulated that AM plays a critical role in the regulation of vascular tone, remodeling and morphogenesis. And although numerous reports have examined the action of AM on cultured vascular cells, the results have not been consistent and have depended on the experimental conditions used. Accordingly, the purpose of this study was to clarify the effect of AM on the proliferation and migration of cultured endothelial cells. Our results revealed that AM promoted the growth and migration of endothelial cells (ECs). AM significantly promoted the proliferation of human umbilical vein endothelial cells (HUVECs) (56.0 +/- 8.7% over the controls at 10( 9) mol/l) and this stimulative effect was inhibited by two AM antagonists, AM(22 52) and calcitonin gene-related peptide (CGRP) (8-37). The number of HUVECs migrated to the lower surface of the transwell apparatus was also increased dose dependently in the AM group (30.4 +/- 4.2% over the controls at 10(-7) mol/l), and this increase was suppressed by the two AM antagonists and by two PKA antagonists, adenosine 3'5'-cyclic monophosphorothioate Rp-isomer and myristoylated protein kinase A inhibitor amide 14-22. The promoting action of AM on endothelial migration was also suppressed by LY294002, an inhibitor for phosphatidylinositol 3-kinase, but not by N(G)-nitro-L-arginine-methyl ester (L NAME), an antagonist for nitric oxide synthase (NOS). These results indicate that AM promotes proliferation and migration of ECs via a cAMP/PKA dependent pathway and lend support to the idea that AM exerts beneficial effects on vascular regeneration and might be used as a novel therapeutic strategy for patients with vascular disease. PMID- 12630818 TI - Adrenomedullin infusion during ischemia/reperfusion attenuates left ventricular remodeling and myocardial fibrosis in rats. AB - Recent studies have demonstrated that the activation of protein kinase Akt attenuates myocardial ischemia/reperfusion injury. However, it remains unknown whether adrenomedullin (AM), which is also a potent Akt activator, has cardioprotective effects after ischemia/reperfusion. In the present study, Sprague-Dawley rats were exposed to a 30-min period of ischemia induced by ligation of the left coronary artery followed by 24-h reperfusion. They were randomized to receive intravenous administration of AM (0.05 microg/kg/min) or saline for 60 min after coronary ligation. We examined the hemodynamics and myocardial apoptosis 24 h after ischemia/reperfusion. Echocardiographic measurements were performed 4 weeks after ischemia/reperfusion. Myocardial infarct size was also measured histologically. AM significantly reduced left ventricular (LV) end-diastolic pressure (17 +/- 2 to 8 +/- 2 mmHg, p < 0.05) and the number of apoptotic nuclei in myocytes (387 +/- 39 to 147 +/- 72 per field, p < 0.05). AM significantly increased LV dP/dt(max) (4,803 +/- 228 to 5,672 +/- 199 mmHg/s, p < 0.05). AM significantly increased LV fractional shortening (23 +/- 2 vs. 28 +/- 2%, p < 0.05), and significantly reduced LV diastolic dimension (7.4 +/- 0.1 to 6.9 +/- 0.1 mm, p < 0.05) and myocardial infarct size (33 +/- 2 to 20 +/- 2%, p < 0.01) 4 weeks after ischemia/reperfusion. In conclusion, AM infusion during ischemia/reperfusion attenuated the development of LV remodeling and myocardial fibrosis in rats. Based on these results, the cardioprotective effects of AM may be attributed at least partly to its anti-apoptotic effect. PMID- 12630819 TI - Targeted disruption of adrenomedullin and alphaCGRP genes reveals their distinct biological roles. AB - Adrenomedullin (AM) and calcitonin gene-related peptide (CGRP) share common structural characteristics and receptors and belong to the same peptide family. Both peptides show a diverse set of biological effects including vasodilation. Recent establishment of gene-knockout mice has revealed the physiological importance of these two peptides. AM -/- mice demonstrated defective vascular formation during embryogenesis and did not survive beyond midgestation. AM +/- heterozygous mice showed high blood pressure and susceptibility to tissue injury. On the other hand, alphaCGRP -/- mice demonstrated elevated peripheral vascular resistance and high blood pressure caused by increased peripheral sympathetic activity. Thus, AM and CGRP have distinct physiological roles. AM is indispensable for normal embryonic development, regulation of blood pressure and tissue protection against injury, whereas alphaCGRP contributes to the regulation of cardiovascular function through inhibitory modulation of sympathetic nervous activity. PMID- 12630820 TI - Organ-protective effects of adrenomedullin. AB - Adrenomedullin (AM), a vasodilatory peptide, has recently been shown to have multipotent properties. Among its other pharmacological actions, AM has been hypothesized to protect organs from hypertension, hypoxia, or infection. In vitro studies have shown that AM has an inhibitory effect on vascular smooth muscle cell proliferation and oxidative stress, but that it enhances nitric oxide (NO) production, which in turn is thought to protect against organ damage. Recent advances in genetic engineering have made it possible to investigate the chronic effects of AM in vivo. Applying genetic engineering, it is revealed that adrenomedullin was shown to protect liver, kidney, vasculature, and heart from septic shock, ischemia and hypertension. However, speculation as to the mechanism of its organ-protective effect varies from report to report. Possible mechanisms include preservation of blood flow, interaction with NO and/or oxidative stress. And although there continue to be technical limitations to the use of these genetically modified models, their application in further investigations should help to clarify the potential efficacy of AM as a new therapeutic agent. PMID- 12630821 TI - Adrenomedullin: a possible autocrine or paracrine hormone in the cardiac ventricles. AB - Adrenomedullin (AM), a potent vasodilator peptide originally isolated from pheochromocytoma, is expressed in cardiovascular tissues such as those of the cardiac atria and ventricles. Cell culture experiments have shown that AM peptide is synthesized and secreted from cardiac myocytes and fibroblasts of neonatal rats. Humoral factors, such as angiotensin II (Ang II) and endothelin-1 (ET-1), and mechanical stress due to pressure and volume overload to the heart have been shown to be involved in AM expression of the myocardium in both in vitro and in vivo studies. The effects of AM on cardiomyocytes and cardiac fibroblasts have been examined in in vitro studies, with the result that AM was shown to exert inhibitory actions on myocyte hypertrophy and on proliferation and collagen production of cardiac fibroblasts in an autocrine or paracrine manner. In rats, experimental therapeutic intervention consisting of transfer of the AM gene or of recombinant AM appears to partly inhibit the progression of cardiac hypertrophy and remodeling. It has been shown that the calcitonin receptor-like receptor (CRLR) and receptor-activity-modifying protein (RAMP) act together to function as AM receptors, although in this regard there are a number of issues, including the cellular mechanism of AM actions, that remain to be addressed. In addition, the role of proadrenomedullin N-terminal 20 peptide (PAMP), which is derived from preproAM, is another topic for future experiments. Collectively, the research data accumulating in this area suggest that AM plays a role as an autocrine or paracrine hormone in the cardiac ventricles, and that AM might be utilized as a therapeutic tool in the treatment of hypertensive or ischemic heart disease. PMID- 12630822 TI - Cardioprotective effect of adrenomedullin in heart failure. AB - Many neurohumoral factors participate in the pathophysiology of heart failure, and adrenomedullin (AM) may be involved in their derangement. This work reviews the accumulating evidence in support of a compensatory role of AM in heart failure, and describes the possible mechanisms of this role. It has been established that plasma AM levels are increased in patients with heart failure in proportion to the severity of the disease. Furthermore, recent studies suggest that plasma AM level is an independent prognostic indicator of heart failure. Thus, AM may be not only a biochemical marker for evaluating the severity of heart failure, but also a prognostic indicator of this syndrome. In patients with heart failure, AM production is increased not only in the plasma, but also in the heart. AM secretion from the failing human heart is also increased, but this increase is small and responds slowly to the stimulus. This phenomenon may be explained by the fact that AM is secreted via a constitutive pathway and that AM is an autocrine and/or a paracrine factor in the heart. An experiment using cultured myocytes suggested that cytokines and mechanical stress are important stimuli for AM production in the heart. Regarding the action of AM in the heart, recent studies have suggested that AM exerts an inotropic action both in vitro and in vivo. AM also attenuates cardiac hypertrophy in myocytes and inhibits proliferation and collagen production in cardiac fibroblasts. These results suggest that AM may be an antifibrotic, antihypertrophic, and positive inotropic factor in the failing and hypertrophied heart. Because AM has many cardiorenal actions, AM administration may be useful for the treatment of heart failure. Indeed, acute administration of AM has been shown to improve the hemodynamics, renal function, and hormonal parameters in patients with heart failure. Moreover, recent studies have shown that AM gene therapy or long-term AM infusion significantly improved cardiac hypertrophy and fibrosis, and prolonged the survival time in an animal model of hypertension and heart failure. In conclusion, these findings suggest that AM plays a compensatory role in the pathophysiology of heart failure and that administration of AM may be a new and promising approach for the treatment of patients with this syndrome. PMID- 12630823 TI - Variations of human adrenomedullin gene and its relation to cardiovascular diseases. AB - The studies concerning the structure and variations of the human adrenomedullin (AM) gene are reviewed, and their relations to the gene function and genetic predisposition to cardiovascular diseases are discussed. The genomic human AM gene is composed of four exons, and the whole nucleotide sequence corresponding to mature AM resides in the fourth exon. In chromosomal sublocalization, the AM gene is located in the distal portion of the short arm of chromosome 11 (11p15.1 3). Analysis of the promoter region of the AM gene has revealed that two transcription factors, nuclear factor for interleukin-6 expression (NF-IL6) and activator protein 2 (AP-2), participate in the regulation of AM gene expression. It is surmised that NF-IL6 mediates inflammatory stimuli and AP-2 mediates signals of phospholipase C and protein kinase C activation. In addition to these factors, hypoxia induces AM gene expression via the hypoxia inducible factor-1 (HIF-1) binding site. The 3'-end of the AM gene is flanked by a microsatellite marker of cytosine adenine (CA) repeats. In Japanese, there are four types of alleles with different CA-repeat numbers: 11, 13, 14 and 19. It is suggested that existence of the 19-repeat allele is associated with genetic predispositions to develop essential hypertension and diabetic nephropathy. PMID- 12630824 TI - Adrenomedullin in heart failure. AB - Patients with heart failure have frequently been reported to show elevated levels of plasma adrenomedullin. These levels generally correlate with severity of hemodynamic dysfunction and also with neurohormonal indices which are activated according to the severity of heart failure. Furthermore, adrenomedullin gene expression in the heart and kidney is increased in experimental and clinical heart failure. A small number of studies have examined the responses to infusion of adrenomedullin in experimental and clinical heart failure. These studies have generally shown that infusion of adrenomedullin has beneficial hemodynamic effects and promotes maintenance or improvement in renal function, although most of these trials were of short duration. The available data suggest that adrenomedullin in the heart, kidney and plasma is increased in heart failure, possibly to counter the activation or actions of vasoconstricting and sodium retaining hormone systems. An improved understanding of the role of adrenomedullin in heart failure might lead to the development of therapeutic agents acting through adrenomedullin receptors. PMID- 12630825 TI - Pulmonary vasodilator response to adrenomedullin in patients with pulmonary hypertension. AB - This study sought to investigate pulmonary vasodilator responses to intrapulmonary and intravenous infusion of adrenomedullin (AM) in patients with pulmonary hypertension. In 10 patients with pulmonary hypertension, blood flow velocity in a segmental pulmonary artery was measured using a Doppler flow wire during intrapulmonary infusion of AM, acetylcholine (ACh), and adenosine triphosphate (ATP). The hemodynamic effects of intravenously administered AM (0.05 microg/kg/min) were examined in another 5 patients with primary pulmonary hypertension. Intrapulmonary infusion of AM, ACh or ATP caused a significant dose dependent increase in blood flow velocity in a segmental pulmonary artery, respectively. The increase in flow velocity with AM at 10(-8) mol/l (41 +/- 6% of the baseline value) was comparable to that with ACh at 10(-4) mol/l (39 +/- 11%) and that with ATP at 10(-5) mol/l (36 +/- 14%), suggesting a strong pulmonary vasodilator activity of AM. Intravenous infusion of AM produced a 41% increase in cardiac index (p < 0.05) and a 30% decrease in pulmonary vascular resistance (p < 0.05) with a 3% reduction in mean pulmonary arterial pressure (p = NS). These results suggest that, on a molar basis, AM may have much more potent pulmonary vasodilator activity than ACh and ATP, and thus may have beneficial hemodynamic effects in patients with pulmonary hypertension. PMID- 12630826 TI - Detection and discrimination of texture modulations defined by orientation, spatial frequency, and contrast. AB - We sought to determine whether the detection and the identification of texture modulations are mediated by a common mechanism. On each trial two textures were presented, one of which contained a modulation in orientation (OM), spatial frequency (FM), or contrast (CM). Observers were required to indicate whether the modulated texture was presented in the first or the second interval as well as the nature of the texture modulation. The results showed that for two of the three pairwise matchings (OM-FM and OM-CM) detection and identification performance were nearly identical, suggesting a common underlying mechanism. However, when FM and CM textures were paired, discrimination thresholds were significantly higher than detection thresholds. In the context of the filter rectify-filter model of texture perception, our results suggest that the mechanisms underlying detection are labeled with respect to their first-order input; i.e., the identities of these mechanisms are available to higher levels of processing. Several possible explanations for the misidentification of FM and CM at detection threshold are considered. PMID- 12630827 TI - Continuous parallax in discrete pixelated integral three-dimensional displays. AB - An evaluation of the retention of continuous parallax in pixelated integral three dimensional image displays is presented. The integral image capture process is first considered, to provide a starting point for the investigation. The complementary display system is then examined in detail. The viewing geometry of the display system is analyzed to provide a foundation for the work to follow, and an experimental investigation and simulations of the characteristics of emitted ray bundles are presented. Next, an analytical model of decoding lenslet array operation is derived, leading to an understanding of the process responsible for production of continuous parallax in replay. It is found that if the lateral resolution of the lenslet is matched to that of the display, continuous parallax is retained in the replayed image, where the finite aberration-limited resolution of the lenslet acts to produce a low-pass reconstruction filter. A condition is derived for optimal continuous parallax in replay, based on a relationship between pixel width and lenslet rms spot size. PMID- 12630828 TI - Experimental determination of object statistics from noisy images. AB - Modern imaging systems rely on complicated hardware and sophisticated image processing methods to produce images. Owing to this complexity in the imaging chain, there are numerous variables in both the hardware and the software that need to be determined. We advocate a task-based approach to measuring and optimizing image quality in which one analyzes the ability of an observer to perform a task. Ideally, a task-based measure of image quality would account for all sources of variation in the imaging system, including object variability. Often, researchers ignore object variability even though it is known to have a large effect on task performance. The more accurate the statistical description of the objects, the more believable the task-based results will be. We have developed methods to fit statistical models of objects, using only noisy image data and a well-characterized imaging system. The results of these techniques could eventually be used to optimize both the hardware and the software components of imaging systems. PMID- 12630829 TI - Ideal-observer computation in medical imaging with use of Markov-chain Monte Carlo techniques. AB - The ideal observer sets an upper limit on the performance of an observer on a detection or classification task. The performance of the ideal observer can be used to optimize hardware components of imaging systems and also to determine another observer's relative performance in comparison with the best possible observer. The ideal observer employs complete knowledge of the statistics of the imaging system, including the noise and object variability. Thus computing the ideal observer for images (large-dimensional vectors) is burdensome without severely restricting the randomness in the imaging system, e.g., assuming a flat object. We present a method for computing the ideal-observer test statistic and performance by using Markov-chain Monte Carlo techniques when we have a well characterized imaging system, knowledge of the noise statistics, and a stochastic object model. We demonstrate the method by comparing three different parallel hole collimator imaging systems in simulation. PMID- 12630830 TI - Relaxed ordered-subset algorithm for penalized-likelihood image restoration. AB - The expectation-maximization (EM) algorithm for maximum-likelihood image recovery is guaranteed to converge, but it converges slowly. Its ordered-subset version (OS-EM) is used widely in tomographic image reconstruction because of its order of-magnitude acceleration compared with the EM algorithm, but it does not guarantee convergence. Recently the ordered-subset, separable-paraboloidal surrogate (OS-SPS) algorithm with relaxation has been shown to converge to the optimal point while providing fast convergence. We adapt the relaxed OS-SPS algorithm to the problem of image restoration. Because data acquisition in image restoration is different from that in tomography, we employ a different strategy for choosing subsets, using pixel locations rather than projection angles. Simulation results show that the relaxed OS-SPS algorithm can provide an order-of magnitude acceleration over the EM algorithm for image restoration. This new algorithm now provides the speed and guaranteed convergence necessary for efficient image restoration. PMID- 12630831 TI - Catchment areas of panoramic snapshots in outdoor scenes. AB - We took panoramic snapshots in outdoor scenes at regular intervals in two- or three-dimensional grids covering 1 m2 or 1 m3 and determined how the root mean square pixel differences between each of the images and a reference image acquired at one of the locations in the grid develop over distance from the reference position. We then asked whether the reference position can be pinpointed from a random starting position by moving the panoramic imaging device in such a way that the image differences relative to the reference image are minimized. We find that on time scales of minutes to hours, outdoor locations are accurately defined by a clear, sharp minimum in a smooth three-dimensional (3D) volume of image differences (the 3D difference function). 3D difference functions depend on the spatial-frequency content of natural scenes and on the spatial layout of objects therein. They become steeper in the vicinity of dominant objects. Their shape and smoothness, however, are affected by changes in illumination and shadows. The difference functions generated by rotation are similar in shape to those generated by translation, but their plateau values are higher. Rotational difference functions change little with distance from the reference location. Simple gradient descent methods are surprisingly successful in recovering a goal location, even if faced with transient changes in illumination. Our results show that view-based homing with panoramic images is in principle feasible in natural environments and does not require the identification of individual landmarks. We discuss the relevance of our findings to the study of robot and insect homing. PMID- 12630832 TI - Kalman filtering for adaptive nonuniformity correction in infrared focal-plane arrays. AB - A novel statistical approach is undertaken for the adaptive estimation of the gain and bias nonuniformity in infrared focal-plane array sensors from scene data. The gain and the bias of each detector are regarded as random state variables modeled by a discrete-time Gauss-Markov process. The proposed Gauss Markov framework provides a mechanism for capturing the slow and random drift in the fixed-pattern noise as the operational conditions of the sensor vary in time. With a temporal stochastic model for each detector's gain and bias at hand, a Kalman filter is derived that uses scene data, comprising the detector's readout values sampled over a short period of time, to optimally update the detector's gain and bias estimates as these parameters drift. The proposed technique relies on a certain spatiotemporal diversity condition in the data, which is satisfied when all detectors see approximately the same range of temperatures within the periods between successive estimation epochs. The performance of the proposed technique is thoroughly studied, and its utility in mitigating fixed-pattern noise is demonstrated with both real infrared and simulated imagery. PMID- 12630833 TI - Study of waveguide grating eigenmodes for unpolarized filtering applications. AB - The dispersion relation of eigenmodes of two-dimensional waveguide gratings is studied with a perturbative model. The analytic expression of the complex wavelength of the modes permits us to predict the shape of the anomalies in the grating reflectivity with respect to the wavelength and the polarization of the incident plane wave. The simultaneous excitation of two independent modes is necessary for obtaining high-efficiency filtering of unpolarized light. We show how this requirement can be met. PMID- 12630834 TI - Application of semiclassical and geometrical optics theories to resonant modes of a coated sphere. AB - This work deals with some aspects of the resonant scattering of electromagnetic waves by a metallic sphere covered by a dielectric layer, in the weak-absorption approximation. We carry out a geometrical optics treatment of the scattering and develop semiclassical formulas to determine the positions and widths of the system resonances. In addition, we show that the mean lifetime of broad resonances is strongly dependent on the polarization of the incident light. PMID- 12630835 TI - Role of the tunneling ray in near-critical-angle scattering by a dielectric sphere. AB - The scattering far zone for light transmitted through a sphere following p - 1 internal reflections by a family of near-grazing incident rays is subdivided into a lit region and a shadow region. The sharpness of the ray theory transition between the lit and the shadow regions is smoothed in wave theory by radiation shed by electromagnetic surface waves. It is shown that when higher-order terms in the physical optics approximation to the phase of the partial-wave scattering amplitudes are included, the transition between the lit and the shadow regions becomes a two-ray-to-zero-ray transition, called a superweak caustic in analogy to the more familiar scattering caustics and weak scattering caustics. One of the merged rays is a tunneling ray. PMID- 12630836 TI - Wave-front sensing by use of a Green's function solution to the intensity transport equation. AB - A method for reconstructing an unknown wave front from measurements of its intensity distribution on two planes along the direction of propagation is described. The method solves the intensity transport equation by use of Neumann boundary conditions, leading to a solution that requires only matrix multiplication. The method provides real-time wave-front reconstruction with high accuracy and is easily reposed to permit reconstruction of the wave front in any orthonormal basis set. PMID- 12630837 TI - Global spectral irradiance variability and material discrimination at Boulder, Colorado. AB - We analyze 7,258 global spectral irradiance functions over 0.4-2.2 microm that were acquired over a wide range of conditions at Boulder, Colorado, during the summer of 1997. We show that low-dimensional linear models can be used to capture the variability in these spectra over both the visible and the 0.4-2.2 microm spectral ranges. Using a linear model, we compare the Boulder data with the previous study of Judd et al. [J. Opt. Soc. Am. 54, 1031 (1964)] over the visible wavelengths. We also examine the agreement of the Boulder data with a spectral database generated by using the MODTRAN 4.0 radiative transfer code. We use a database of 223 minerals to consider the effect of the spectral variability in the global spectral irradiance functions on hyperspectral material identification. We show that the 223 minerals can be discriminated accurately over the variability in the Boulder data with subspace projection techniques. PMID- 12630838 TI - Eigenfunctions of the offset Fourier, fractional Fourier, and linear canonical transforms. AB - The offset Fourier transform (offset FT), offset fractional Fourier transform (offset FRFT), and offset linear canonical transform (offset LCT) are the space shifted and frequency-modulated versions of the original transforms. They are more general and flexible than the original ones. We derive the eigenfunctions and the eigenvalues of the offset FT, FRFT, and LCT. We can use their eigenfunctions to analyze the self-imaging phenomena of the optical system with free spaces and the media with the transfer function exp[j(h2x2 + h1x + h0)] (such as lenses and shifted lenses). Their eigenfunctions are also useful for resonance phenomena analysis, fractal theory development, and phase retrieval. PMID- 12630839 TI - Importance of the phase and amplitude in the fractional Fourier domain. AB - The importance of the amplitude and phase in the fractional Fourier transform (FT) domain is analyzed on the basis of the rectangular signal and the real-world image. The quality of signal restoration from only the amplitude or from only the phase of its fractional FT by applying the inverse fractional FT is considered. It is shown that the signal reconstructed from the amplitude of the fractional FT usually reveals the main features of the original signal only for relatively low fractional orders. On the basis of phase information in the fractional FT domains, significant details of the signal can be obtained for nearly all fractional orders. PMID- 12630840 TI - Theory of total-internal-reflection tomography. AB - A method is presented to reconstruct three-dimensional tomographic images of weakly scattering objects with subwavelength resolution. The method may be applied to data available in phase-sensitive, total-internal-reflection microscopy. The results follow from an analysis of the near-field inverse scattering problem with evanescent waves. PMID- 12630841 TI - Guided modes in a uniaxial multilayer. AB - An algorithm is presented for simulation of guided modes in a multilayer uniaxial structure with each layer characterized by its own ellipsoid of refractive indices and direction of optical axis. The proposed approach is based on presenting an electromagnetic field in each layer as a linear combination of ordinary and extraordinary waves coupled through the boundary conditions. The problem is reduced to two dimensions by considering the waves with a given projection of the wave vector on the plane of the waveguide. No a priori assumption about the guided-mode polarization is required in this method. Hybrid polarized modes appear naturally as solutions of a system of linear equations with respect to the amplitudes of the ordinary and extraordinary waves. The proposed approach covers a wide variety of important practical cases including isotropic waveguides, surface waves at the boundary between positive uniaxial crystal and isotropic medium, surface plasmons at metallic interfaces, uniaxial multilayers in a very general form, and leaky modes in such structures. PMID- 12630842 TI - Adaptive Hermite-Gauss decomposition method to analyze optical dielectric waveguides. AB - A novel spectral method with variable transformation, the adaptive Hermite-Gauss decomposition method (A-HGDM), has been developed and applied to the analysis of three-dimensional (3D) dielectric structures. The proposed method includes an optimization strategy to automatically find the quasi-optimum numerical parameters of the variable transformation with low computational effort. The technique has been tested by analyzing two typical 3D dielectric structures: the rectangular step-index waveguide and the rib-waveguide directional coupler. In both cases, the A-HGDM increases the accuracy of the Hermite-Gauss decomposition method (HGDM), especially when the mode is near cutoff, and improves the computational efficiency of previously published optimization strategies (optimized HGDM). PMID- 12630843 TI - Temporal coupled-mode theory for the Fano resonance in optical resonators. AB - We present a theory of the Fano resonance for optical resonators, based on a temporal coupled-mode formalism. This theory is applicable to the general scheme of a single optical resonance coupled with multiple input and output ports. We show that the coupling constants in such a theory are strongly constrained by energy-conservation and time-reversal symmetry considerations. In particular, for a two-port symmetric structure, Fano-resonant line shape can be derived by using only these symmetry considerations. We validate the analysis by comparing the theoretical predictions with three-dimensional finite-difference time-domain simulations of guided resonance in photonic crystal slabs. Such a theory may prove to be useful for response-function synthesis in filter and sensor applications. PMID- 12630844 TI - Noncanonical vortex transformation and propagation in a two-dimensional optical system. AB - We produced an axial and canonical optical vortex by using a computer-generated hologram and then converted it to a noncanonical vortex by passing it through a cylindrical lens. We conducted an experimental study of the shape and trajectory of the noncanonical vortex as it propagates in free space and obtained an analytical expression explaining our experimental results. The computed trajectory and shape of the noncanonical vortex agree quite well with our experimental results. PMID- 12630845 TI - Anisotropic pure-phase plates for quality improvement of partially coherent, partially polarized beams. AB - From a theoretical point of view, the use of anisotropic pure-phase plates (APP) is considered in order to improve the quality parameter of certain partially coherent, partially polarized beams. It is shown that, to optimize the beam quality parameter, the phases of the two Cartesian components of the field at the output of the APP plate should be identical and should exhibit a quadratic dependence on the radial polar coordinate. PMID- 12630846 TI - Propagation properties of ultrashort pulsed bessel beams in dispersive media. AB - A detailed study of ultrashort pulsed Bessel beams in linear dispersive media is performed. The spatial and temporal parts of pulsed Jn beams are separable in dispersive media, provided that the parameter a is independent of frequency omega. The spatial part keeps the Jn shape unchanged during propagation. The temporal evolution behavior of pulsed Jn beams depends on the material's dispersion and diffraction. The pulses can be broadening and become negatively chirped while propagating in anomalous dispersive media. In normal dispersive media, the pulses can be broadening and positively or negatively chirped; even dispersion-free propagation can be achieved if the beam and material parameters are suitably chosen. The condition under which higher-order dispersive effects can be neglected is also discussed. PMID- 12630847 TI - Soft x-ray holographic computerized tomography imaging: experimental research. AB - A high-resolution three-dimensional (3D) imaging technology has been developed, which is a combination of x-ray holography and computerized tomography (CT) technology called holographic computerized tomography (HCT). The theory and experimental techniques on biological specimens with the use of synchrotron radiation are discussed. Projections at different angles are reconstructed with the numerical method of in-line holography, and then the reconstructed data with a higher lateral resolution are used to restore the 3D image by the CT technique. With this method, the degradation caused by the diffraction of x rays is canceled, and 3D images with high resolution of micrometer magnitude in both the lateral and the longitudinal directions are obtained. PMID- 12630849 TI - The hydrothermal synthesis of zeolites: history and development from the earliest days to the present time. PMID- 12630848 TI - Biochemical modulation of Cisplatin mechanisms of action: enhancement of antitumor activity and circumvention of drug resistance. PMID- 12630851 TI - Carbon-carbon bond forming reactions mediated by silicon Lewis acids. PMID- 12630850 TI - Cyclooxygenase (COX) inhibitors: a comparative QSAR study. PMID- 12630852 TI - Principles governing Mg, Ca, and Zn binding and selectivity in proteins. PMID- 12630853 TI - Persistent and stable radicals of the heavier main group elements and related species. PMID- 12630854 TI - Recent advances in the Baylis-Hillman reaction and applications. PMID- 12630855 TI - The combinatorial synthesis of bicyclic privileged structures or privileged substructures. PMID- 12630856 TI - Bioconjugation by copper(I)-catalyzed azide-alkyne [3 + 2] cycloaddition. AB - The copper-catalyzed cycloaddition reaction between azides and alkynes functions efficiently in aqueous solution in the presence of a tris(triazolyl)amine ligand. The process has been employed to make rapid and reliable covalent connections to micromolar concentrations of protein decorated with either of the reactive moieties. The chelating ligand plays a crucial role in stabilizing the Cu(I) oxidation state and protecting the protein from Cu(triazole)-induced denaturation. Because the azide and alkyne groups themselves are unreactive with protein residues or other biomolecules, their ligation is of potential utility as a general bioconjugation method. PMID- 12630857 TI - [(Fe(III)(OH(2))(2))(3)(A-alpha-PW(9)O(34))(2)](9-) on cationic silica nanoparticles, a new type of material and efficient heterogeneous catalyst for aerobic oxidations. AB - Polyoxometalates (POMs) electrostatically bind to silica nanoparticles coated with cationic aluminum oxide "(Si/AlO2)n+" to form a new type of material (the anionic POMs replace Cl- counterions associated with the cationic surface sites). Association of a new approximately D3h POM of formula [(FeIII(OH2)2)3(A-alpha PW9O34)2]9- (1) with the cationic nanoparticles (to form "K81/(Si/AlO2)") was studied in detail. Elemental analysis, particle sizes from both laser light scattering and TEM before and after association of 1, the size of 1 from X-ray crystallography, and other methods provide mutually consistent data that indicate about 58 K8[(FeIII(OH2)2)3(A-alpha-PW9O34)2]- monoanions associate with the average nanoparticle (diameter of the K81/(Si/AlO2) product = approximately 17 nm). While heterogeneity of the cationic sites and roughness of the (Si/AlO2)n+ surfaces make the associated POMs structurally nonuniform, the equivalent of approximately 1 monolayer of 1 is present in K81/(Si/AlO2). Remarkably, while 1, the precursor (Si/AlO2)n+, and the components of 1, each alone, are inactive as catalysts for O2/air-based oxidation of sulfides or aldehydes in solution, K81/(Si/AlO2) is an active catalyst for both reactions (facile reaction with air at low temperature). PMID- 12630858 TI - Nanostructured anisotropic ion-conductive films. AB - A flexible self-standing film with layered nanostructures was obtained by in situ photopolymerization of a new smectic liquid-crystalline monomer containing a tetra(oxyethylene) moiety, which forms a macroscopically oriented complex with lithium salts. The resultant films show two-dimensional ionic conductivity. PMID- 12630859 TI - Structure and mechanism of action of isopentenylpyrophosphate dimethylallylpyrophosphate isomerase. AB - We have obtained the three-dimensional X-ray crystallographic structure of a C67A mutant Escherichia coli isopentenylpyrophosphate-dimethylallylpyrophosphate isomerase (EC 5.3.3.2) complexed with the bromohydrin of isopentenylpyrophosphate, at 1.93 A resolution. The overall backbone fold is very similar to that obtained previously for the wild-type enzyme in the presence of a divalent metal cation (Mn2+ or Mg2+). However, in the new structure, there are two metal binding sites, not just one. The first metal binding site is occupied by Mn2+, coordinated to three histidine and two glutamate residues, while the second is occupied by Mg2+, coordinated to two bromohydrin-ligand phosphate oxygens, the carbonyl oxygen of A67, a carboxyl oxygen of E87, and two water molecules. The C3 hydroxyl group of the bromohydrin inhibitor is involved in a short hydrogen bond to the carboxyl group of E116, one of the two Mn-bound glutamates. The structure obtained is consistent with a mechanism of action of the enzyme in which the carboxyl group of E116 protonates the double bond in isopentenylpyrophosphate, forming a carbocation, followed by removal of a C2 proton by the thiolate of C67, in the wild-type enzyme. The inhibition of the enzyme by a wide variety of other potent inhibitors is also readily explained on the basis of the bromohydrin inhibitor structure. PMID- 12630860 TI - Peptides derived from two dynamically disordered proteins self-assemble into amyloid-like fibrils. AB - Short peptides derived from p14ARF and Hdm2 (14 and 15 amino acids in length, respectively), two cancer associated proteins, have been found to co-assemble into amyloid-like structures. Larger protein domains containing these peptide segments interact in cells and also undergo a disorder-to-order transition upon binding in vitro. In contrast to the association of beta-strand assemblies with amyloid diseases, the system described herein utilizes the formation of binary, extended beta-strands as a novel mechanism of biomolecular assembly. The beta strand-containing fibrils formed from these peptides may allow the directed assembly of decorated fibrils with applications as biological nanostructures. PMID- 12630861 TI - Ligand effects on charge transport in platinum(II) acetylides. AB - To investigate the electrical characteristics of organometallic complexes as molecular conductors, organometallic pi-conjugated molecules of the type trans [PtL2(CCC6H4SAc-4)2], where L = PCy3, PBu3, PPh3, P(OEt)3, P(OPh)3, were synthesized and characterized by NMR, IR, UV, and X-ray spectroscopies. For the three complexes (L = PCy3, PPh3, and P(OEt)3) that could be measured using a cross-wire junction technique, the current-voltage (I-V) characteristics of a molecular monolayer of these complexes showed no ligand effect, despite spectroscopic evidence that electronic interaction between the phosphine ligands and the pi-system does occur. It was concluded that the tunneling efficiency across the molecule is the determining factor for conduction in this metal molecule-metal system. It was also shown that the incorporation of a transition metal in pi-conjugated molecular wires does not adversely affect charge transport compared to all-carbon pi-conjugated molecular wires. PMID- 12630862 TI - Isomeric forms of divalent heavier group 14 element hydrides: characterization of Ar'(H)GeGe(H)Ar' and Ar'(H)(2)GeGeAr'.PMe(3) (Ar' = C(6)H(3)-2,6-Dipp(2); Dipp = C(6)H(3)-2,6-Pr(i)(2)). AB - The reduction of Ar'GeCl (Ar' = C6H3-2,6-Dipp2; Dipp = C6H3-2,6-Pri2) with LiBH(Bus)3 affords the first heavier group 14 element dimetallene hydride Ar'(H)GeGe(H)Ar' which, upon further reaction with PMe3, yields the base stabilized isomeric form Ar'(H)2GeGeAr'.PMe3. PMID- 12630863 TI - Selective stabilization of the chorismate mutase transition state by a positively charged hydrogen bond donor. AB - Citrulline was incorporated via chemical semisynthesis at position 90 in the active site of the AroH chorismate mutase from Bacillus subtilis. The wild-type arginine at this position makes hydrogen-bonding interactions with the ether oxygen of chorismate. Replacement of the positively charged guanidinium group with the isosteric but neutral urea has a dramatic effect on the ability of the enzyme to convert chorismate into prephenate. The Arg90Cit variant exhibits a >104-fold decrease in the catalytic rate constant kcat with a 2.7-fold increase in the Michaelis constant Km. In contrast, its affinity for a conformationally constrained inhibitor molecule that effectively mimics the geometry but not the dissociative character of the transition state is only reduced by a factor of approximately 6. These results show that an active site merely complementary to the reactive conformation of chorismate is insufficient for catalysis of the mutase reaction. Instead, electrostatic stabilization of the polarized transition state by provision of a cationic hydrogen bond donor proximal to the oxygen in the breaking C-O bond is essential for high catalytic efficiency. PMID- 12630864 TI - Unfunctionalized, alpha-epimerizable nonracemic ketones and aldehydes can be accessed by crystallization-induced dynamic resolution of imines. AB - This paper describes an operationally simple deracemization process of aldehydes and ketones. This new crystallization-induced dynamic resolution (CIDR) protocol allows for nearly complete conversion of the racemic mixture into one enantiomer. Crystallization of imines derived from racemic ketones or aldehydes 1 and trans (1R,2R)-1-amino-6-nitroindan-2-ol (2) afforded diastereomerically pure, crystalline imines 3. Biphasic hydrolysis of 3 then affords recovered 2 and enantiomerically enriched 1 in high yield and er (substrate, yield/ee: 2 methylcyclohexanone, 97%/92; 2-ethylhexanal, 94%/98; 2-methylcyclopentanone, 94%/98; 2-cyclohexylcyclohexanone, ND/98; 3-methyl-2-pentanone, ND/76). The scope, limitations, and industrial perspective of this process are discussed. This highly effective CIDR process is likely due to pi-stacking of 2 and a hydrogen bonding of the imine with the free hydroxyl of 2 in the solid state. PMID- 12630865 TI - A one-pot diastereoselective synthesis of cis-3-hexene-1,6-diols via an unusually reactive organozinc intermediate. AB - A highly diastereoselective method for the synthesis of cis-3-hexene-1,6-diols has been developed. This new reaction proceeds with excellent control of diastereoselectivity over four stereocenters and the double bond geometry. The diols are made in a one-pot procedure involving hydroboration of a terminal alkyne and transmetalation to zinc to give a divinylzinc intermediate. This intermediate undergoes reductive elimination, forming a C=C bond. In the absence of a trapping reagent, diene is liberated (70% yield); however, in the presence of ketones or aldehydes, the proposed intermediate metallocyclopentene is trapped via a double insertion of the carbonyl substrate. Workup provides the diols in 47 86% yield. PMID- 12630866 TI - Radical anion of isolable dialkylsilylene. AB - By the reduction of an isolable dialkylsilylene, 2,2,5,5 [tetrakis(trimethylsilyl)]-1-silacyclopentane-1,1-diyl (1), with cesium, rubidium, potassium, sodium, and lithium 4,4'-di(tert-butyl)biphenylide in DME at low temperatures, the corresponding silylene radical anion 2 was generated as the first persistent silylene radical anion in solution and characterized by ESR spectroscopy. Radical anion 2 is rather stable at -70 degrees C in DME but decomposes rapidly at room temperature with a half-life time of ca. 20 min. The g factor and 29Si hyperfine splitting constants (hfs's) of 2 are almost independent of the countercations, indicating that 2 exists as a free ion or a solvent separated ion pair in a polar DME solution. A very small hfs due to the 29Si nucleus of the divalent silicon (3.0 mT) as well as a very large g-factor (2.0077) indicates that an unpaired electron is accommodated in the vacant 3ppi orbital of silylene 1. PMID- 12630868 TI - Diastereoselective addition of alcohol to diastereotopic silylenes. AB - The successful example of diastereoselective addition of alcohol to diastereotopic silylenes is reported. PMID- 12630867 TI - Electrochemical coding technology for simultaneous detection of multiple DNA targets. AB - Nucleic-acid hybridization assays based on the use of different inorganic-colloid (quantum dots) nanocrystal tracers for the simultaneous electrochemical measurements of multiple DNA targets are described. Three encoding nanoparticles (zinc sulfide, cadmium sulfide, and lead sulfide) are used to differentiate the signals of three DNA targets in connection to stripping-voltammetric measurements of the heavy metal dissolution products. These products yield well-defined and resolved stripping peaks at -1.12 V (Zn), -0.68 V (Cd), and -0.53 V (Pb) at the mercury-coated glassy-carbon electrode (vs Ag/AgCl reference). The position and size of these peaks reflect the identity and level of the corresponding DNA target. The multi-target detection capability is coupled to the amplification feature of stripping voltammetry (to yield femtomole detection limits) and with an efficient magnetic removal of nonhybridized nucleic acids to offer high sensitivity and selectivity. The protocol is illustrated for the simultaneous detection of three DNA sequences related to the BCRA1 breast-cancer gene in a single sample in connection to magnetic beads bearing the corresponding oligonucleotide probes. The new electrochemical coding is expected to bring new capabilities for DNA diagnostics, and for bioanalysis, in general. PMID- 12630869 TI - (Z)-alpha-haloacrylates: an exceptionally stereoselective preparation via Cr(II) mediated olefination of aldehydes with trihaloacetates. AB - (Z)-alpha-Fluoro-, (Z)-alpha-chloro-, and (Z)-alpha-bromoacrylates were obtained with unprecedented yield and stereocontrol (>99%) via addition of the corresponding commercial trihaloacetates to aldehydes at room temperature using stoichiometric Cr(II) salts or catalytic Cr(II) with a regeneration system. The intermediate 2,2-dihalo-3-hydroxy adducts could be isolated in good yields under conditions of limited reagent at 0 degrees C. PMID- 12630870 TI - Asymmetric skeletal rearrangement of symmetrically alpha,alpha-disubstituted alpha-amino aldehydes: a new entry to optically active alpha-hydroxy ketones. AB - A unique asymmetric skeletal rearrangement of symmetrically alpha,alpha disubstituted alpha-amino aldehydes has been accomplished for the first time using a chiral organoaluminum Lewis acid 1. For instance, treatment of (S)-2,2' bis(trifluoromethanesulfonylamino)-1,1'-binaphthyl with Me3Al (1.0 equiv) in toluene at room temperature for 15 min and at 110 degrees C for an additional 15 min produced (S)-1, and a subsequent reaction with alpha -amino aldehyde 2a (R = CH2Ph) at -78 degrees C for 4 h and at -40 degrees C for 12 h resulted in the smooth rearrangement to the zwitterionic iminium intermediate A, which furnished the alpha-hydroxy ketone 3a (R = CH2Ph) in 93% isolated yield with 95% ee (S) after acidic hydrolysis. This result, together with other representative examples, clearly demonstrates the effectiveness of the present method for the hitherto difficult asymmetric synthesis of acyloins. Furthermore, we found that the treatment of the in situ generated A with DIBAH afforded the corresponding anti amino alcohol exclusively without loss of enantiomeric excess. Our approach casts light on the previously unexplored yet potential utility of alpha-amino aldehydes as synthetic building blocks and also provides a new entry to optically active alpha-hydroxy ketones and 1,2-amino alcohols. PMID- 12630871 TI - Coordination of alkenes and alkynes to a cationic d(0) zirconocene alkoxide complex. AB - This paper describes the synthesis of base-free (C5R5)2Zr(OtBu)+ cations, the direct observation of nonchelated alkene and alkyne adducts of these cations, and studies of the thermodynamic and dynamic properties of these novel species. Reaction of %@mt;sys@%Cp%@/xs;55;%lnwidth@%'%@/xs;63;(%lnwidth-x55)@%%@mh; x63@%%@sb@%2%@sbx@%%@/hd@%ZrMe%@sb@%2%@sbx@%%@mx@% (Cp' = C5H4Me) with tert-butyl alcohol followed by [Ph3C][B(C6F5)4] in benzene yields [%@mt;sys@%Cp%@/xs;55;%lnwidth@%'%@/xs;63;(%lnwidth-x55)@%%@mh; x63@%%@sb@%2%@sbx@%%@/hd@%Zr%@/hd@%%@fn;(;vis;full;auto@%O%@ital@%%@ex@%t%@rsf@%% exx@%%@/hd@%Bu%@fnx;);vis;full@%%@/hd@%%@mx@% ][B(C6F5)4] (1), which exists as %@mt;sys@%Cp%@/xs;55;%lnwidth@%'%@/xs;63;(%lnwidth-x55)@%%@mh; x63@%%@sb@%2%@sbx@%%@/hd@%Zr%@/hd@%%@fn;(;vis;full;auto@%O%@ital@%%@ex@%t%@rsf@%% exx@%%@/hd@%Bu%@fnx;);vis;full@%%@/hd@%%@fn;(;vis;full;auto@%ClR%@fnx;);vis;full@ %@ex@%+%@exx@%%@mx@% solvent adducts in C6D5Cl and CD2Cl2 solutions. Addition of ligands L (L = ethylene, propylene, propyne, 2-butyne, CO, phenylacetylene, allene, 1-hexene, cis-2-butene) to 1 in CD2Cl2 at -89 degrees C results in reversible formation of %@mt;sys@%Cp%@/xs;55;%lnwidth@%'%@/xs;63;(%lnwidth x55)@%%@mh; x63@%%@sb@%2%@sbx@%%@/hd@%Zr%@/hd@%%@fn;(;vis;full;auto@%O%@ital@%%@ex@%t%@rsf@%% exx@%%@/hd@%Bu%@fnx;);vis;full@%%@/hd@%%@fn;(;vis;full;auto@%L%@fnx;);vis;full@%% ex@%+%@exx@%%@mx@% adducts. NMR data for %@mt;sys@%Cp%@/xs;55;%lnwidth@%'%@/xs;63;(%lnwidth-x55)@%%@mh; x63@%%@sb@%2%@sbx@%%@/hd@%Zr%@/hd@%%@fn;(;vis;full;auto@%O%@ital@%%@ex@%t%@rsf@%% exx@%%@/hd@%Bu%@fnx;);vis;full@%%@/hd@%%@fn;(;vis;full;auto@%H%@sb@%2%@sbx@%C=%@/ d@%CHMe%@fnx;);vis;full@%%@ex@%+%@exx@%%@mx@% (4) indicate that the propylene coordinates unsymmetrically and is polarized with positive charge buildup at Cint. Equilibrium constants, defined by Keq = [Zr-L][1]-1[L]-1, vary in the order CO > propyne > 2-butyne > phenylacetylene > ethylene > allene > propylene > 1 hexene > cis-2-butene > vinyl chloride. Loss of L from %@mt;sys@%Cp%@/xs;55;%lnwidth@%'%@/xs;63;(%lnwidth-x55)@%%@mh; x63@%%@sb@%2%@sbx@%%@/hd@%Zr%@/hd@%%@fn;(;vis;full;auto@%O%@ital@%%@ex@%t%@rsf@%% exx@%%@/hd@%Bu%@fnx;);vis;full@%%@/hd@%%@fn;(;vis;full;auto@%L%@fnx;);vis;full@%% ex@%+%@exx@%%@mx@% to give 1 appears to proceed via associative displacement by CD2Cl2 in most cases. PMID- 12630872 TI - Evidence for substrate activation of electron transfer from methylamine dehydrogenase to amicyanin. AB - Electron transfer (ET) from methylamine dehydrogenase (MADH) to amicyanin may be true or gated ET, depending upon the redox form of MADH. ET from the substrate reduced aminoquinol form of MADH is gated, and the reaction rate is dependent on the presence of monovalent cations. This ET reaction has been studied in buffer free of monovalent cations. The reaction rate is orders of magnitude less than with saturating concentrations of monovalent cation. Analysis of the temperature dependence of this slow reaction, however, reveals that it is a true ET reaction. The rate of MADH reduction by substrate and the steady-state rate of substrate dependent reduction of amicyanin by MADH were examined in different buffers. The results reveal that, in the steady state, the protonated methylammonium substrate performs the role previously attributed to monovalent cations in regulating the rate and mechanism of ET from MADH. The two putative cation binding sites previously observed in the crystal structure of MADH may now be assigned distinct roles, one as a catalytic substrate binding site and the other as a noncatalytic regulatory substrate binding site. PMID- 12630873 TI - Inert site in a protein zinc cluster: isotope exchange by high resolution mass spectrometry. AB - Using electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry, we show that one Zn2+ ion in the Zn4 cluster of cyanobacterial metallothionein is inert to exchange with 67Zn2+. We suggest that this is zinc in site A, which together with the surrounding alpha and beta secondary structure forms a zinc-finger fold. PMID- 12630874 TI - The first stable mononuclear silyl palladium hydrides. AB - The reaction of tertiary silanes with the low valent palladium complex [(mu dcpe)Pd]2 affords equilibrium mixtures with mononuclear silyl palladium hydrides. These complexes have been characterized by NMR spectroscopy and, in one case, by X-ray crystallography. The silyl palladium hydride complexes rapidly interchange silicon and hydride coordination environments in solution which give rise to extraordinary temperature-dependent kinetic isotope effects for the fluxional process. An intermediate 2eta-Si-H complex is proposed for the interchange. PMID- 12630875 TI - The synthesis of a dicyclopenta[a,e]pentalene via a molybdenum hexacarbonyl mediated tandem allenic Pauson-Khand reaction. AB - An approach to the 14pi annulene has been developed, by employing the molybdenum hexacarbonyl-mediated tandem allenic Pauson-Khand reaction as a key step. A di triisopropylsilyl substituted dicyclopenta[a,e]pentalene was synthesized. This 14pi annulene showed some electronic delocalization, which was not observed in previous cases. PMID- 12630876 TI - A mild, catalytic, and highly selective method for the oxidation of alpha,beta enones to 1,4-enediones. AB - A new and simple method is described for the one-step oxidation of alpha,beta enones to 1,4-enediones in good yields (generally 80-90%) using t butylhydroperoxide as stoichiometric oxidant and 20% Pd(OH)2 on carbon (5 mol %) as catalyst in CH2Cl2 solution. The same reagents have been used to convert ethylene ketals of alpha,beta-enones to the corresponding monoethylene ketals of 1,4-enediones. Seven representative examples are presented in Table 1. All of the available evidence on these oxidations points to a radical-chain mechanism initiated by the tert-butylperoxy radical (see Scheme 1). gamma-tert-Butylperoxy ethers are formed as major products in the oxidation of alpha,beta-enones possessing only a single gamma-hydrogen. PMID- 12630877 TI - Hydrosilylation of a dinuclear tantalum dinitrogen complex: cleavage of N2 and functionalization of both nitrogen atoms. AB - Hydrosilylation of the ditantalum dinitrogen complex ([NPN]Ta)2(mu-H)2(mu eta1:eta2-N2) proceeds via an addition reaction to produce ([NPN]TaH)(mu-H)2(mu eta1:eta2-N-NSiH2Bu)(Ta[NPN]), which contains a new N-Si bond and a terminal tantalum hydride; this species has been characterized by NMR spectroscopy and X ray diffraction. This complex undergoes reductive elimination of H2 followed by N N bond cleavage to generate a new intermediate with the formula ([NPN]TaH)(mu N)(mu-NSiH2Bu)(Ta[NPN]); confirmation of N-N bond cleavage is evident from the 15N-labeled isotopomer that displays an absence of 15N-15N scalar coupling in the 15N NMR spectrum. In the presence of additional silane, a second hydrosilylation and reductive elimination results to give ([NPN]Ta)2(mu-NSiH2Bu)2, a species in which each dinitrogen-derived N atom has been converted to a bridging silylimide ligand. This latter complex displays C2h symmetry both in solution and in the solid state. PMID- 12630878 TI - Application of Bronsted-type LFER in the study of the phospholipase C mechanism. AB - Phosphatidylinositol-specific phospholipase C cleaves the phosphodiester bond of phosphatidylinositol to form inositol 1,2-cyclic phosphate and diacylglycerol. This enzyme also accepts a variety of alkyl and aryl inositol phosphates as substrates, making it a suitable model enzyme for studying mechanism of phosphoryl transfer by probing the linear free-energy relationship (LFER). In this work, we conducted a study of Bronsted-type relationship (log k = beta(lg) pK(a) + C) to compare mechanisms of enzymatic and nonenzymatic reactions, confirm the earlier proposed mechanism, and assess further the role of hydrophobicity in the leaving group as a general acid-enabling factor. The observation of the high negative Bronsted coefficients for both nonenzymatic (beta(lg) = -0.65 to -0.73) and enzymatic cleavage of aryl and nonhydrophobic alkyl inositol phosphates (beta(lg) = -0.58) indicates that these reactions involve only weak general acid catalysis. In contrast, the enzymatic cleavage of hydrophobic alkyl inositol phosphates showed low negative Bronsted coefficient (beta(lg) = -0.12), indicating a small amount of the negative charge on the leaving group and efficient general acid catalysis. Overall, our results firmly support the previously postulated mechanism where hydrophobic interactions between the enzyme and remote parts of the leaving group induce an unprecedented negative-charge stabilization on the leaving group in the transition state. PMID- 12630879 TI - Cavity-directed synthesis within a self-assembled coordination cage: highly selective [2 + 2] cross-photodimerization of olefins. AB - Herein reported is the highly selective [2 + 2] cross-photodimerization of olefins within a self-assembled coordination cage that acts as a molecular flask in an aqueous medium. An M(6)L(4) coordination nanocage that self-assembles from six Pd(II) complexes and four tridentate ligands accommodates two different kinds of large olefin molecules such as acenaphthylene and 5-ethoxynaphthoquinone in a pairwise selective fashion. This prerequisite recognition mode makes possible the selective [2 + 2] cross-photodimerization of the olefins within the cavity. The reaction is extremely efficient in terms of reaction rate, stereoselectivity, and, most importantly, pairwise selectivity. Maleimide derivatives, which themselves are photochemically inactive under ordinary conditions, are also cross dimerized with acenaphthylene or dibenzosuberenon quite efficiently. These results are in sharp contrast to those of common photodimerization in organic media, where the yields and selectivities are generally poor to moderate. The key step of the exclusive formation of the cross-dimers stems from the selective formation of ternary complexes before irradiation, which is governed by the size compatibility of the guests with the confined space of the cavity. PMID- 12630881 TI - Amphiphilic Janus micelles with polystyrene and poly(methacrylic acid) hemispheres. AB - We describe the synthesis and the solution properties of Janus micelles containing a polybutadiene (PB) core and a compartmentalized corona consisting of a poly(methacrylic acid) (PMAA) and a polystyrene (PS) hemisphere. The Janus micelles were obtained via cross-linking the middle block of a microphase separated PS-block-PB-block-PMMA triblock copolymer in the bulk state, followed by alkaline hydrolysis of the poly(methyl methacrylate) (PMMA) ester groups. Results of fluorescence correlation spectroscopy, field flow fractionation, light scattering, cryogenic transmission electron microscopy, scanning electron microscopy, and scanning force microscopy indicate that above a critical aggregation concentration of about 0.03 g/L spherical supermicelles are formed from about 30 PS-PMAA micelles in aqueous solution in the presence of NaCl. These supermicelles have radii of 40-60 nm, significantly increasing on ionization (pH >6). In addition, very large spherical objects are observed with radii of 100-250 nm. PMID- 12630880 TI - Trimethyl phosphite as a trap for alkoxy radicals formed from the ring opening of oxiranylcarbinyl radicals. Conversion to alkenes. Mechanistic applications to the study of C-C versus C-O ring cleavage. AB - Trimethyl phosphite, (MeO)(3)P, is introduced as an efficient and selective trap in oxiranylcarbinyl radical (2) systems, formed from haloepoxides 8-13 under thermal AIBN/n-Bu(3)SnH conditions at about 80 degrees C. Initially, the transformations of 8-13, in the absence of phosphite, to allyl alcohol 7 and/or vinyl ether 5 were measured quantitatively (Table 1). Structural variations in the intermediate oxiranylcarbinyl (2), allyloxy (3), and vinyloxycarbinyl (4) radicals involve influences of the thermodynamics and kinetics of the C-O (2 --> 3, k(1)) and C-C (2 --> 4, k(2)) radical scission processes and readily account for the changes in the amounts of product vinyl ether (5) and allyl alcohol (7) formed. Added (MeO)(3)P is inert to vinyloxycarbinyl radical 4 and selectively and rapidly traps allyloxy radical 3, diverting it to trimethyl phosphate and allyl radical 6. Allyl radicals (6) dimerize or are trapped by n-Bu(3)SnH to give alkenes, formed from haloepoxides 8, 9, and 13 in 69-95% yields. Intermediate vinyloxycarbinyl radicals (4), in the presence or absence of (MeO)(3)P, are trapped by n-Bu(3)SnH to give vinyl ethers (5). The concentrations of (MeO)(3)P and n-Bu(3)SnH were varied independently, and the amounts of phosphate, vinyl ether (5), and/or alkene from haloepoxides 10, 11, and 13 were carefully monitored. The results reflect readily understood influences of changes in the structures of radicals 2-4, particularly as they influence the C-O (k(1)) and C-C (k(2)) cleavages of intermediate oxiranylcarbinyl radical 2 and their reverse (k( 1), k(-2)). Diversion by (MeO)(3)P of allyloxy radicals (3) from haloepoxides 11 and 12 fulfills a prior prediction that under conditions closer to kinetic control, products of C-O scission, not just those of C-C scission, may result. Thus, for oxiranylcarbinyl radicals from haloepoxides 11, 12, and 13, C-O scission (k(1), 2 --> 3) competes readily with C-C cleavage (k(2), 2 --> 4), even though C-C scission is favored thermodynamically. PMID- 12630882 TI - Twisted amide reduction under Wolff-Kishner conditions: synthesis of a benzo-1 aza-adamantane derivative. AB - A synthesis of 4,5-benzo-1-aza-tricyclo[4.3.1.1(3,8)]undecane (1), a benzo-1-aza adamantane derivative, is described and features a previously unknown application of the Wolff-Kishner reduction of a nonresonance stabilized or "twisted" amide. An intermediate amino ester is converted to a severely "twisted amide", which, when exposed to hydrazine in alcohol, provides the corresponding "twisted" amino hydrazone. Wolff-Kishner conditions (KOH/ethylene glycol, 200 degrees C) provide the reduced target 1 without hydrolysis to amino acid derivatives. These operations are conveniently performed in a single flask in high yield. PMID- 12630883 TI - Charge-transfer mechanism for electrophilic aromatic nitration and nitrosation via the convergence of (ab initio) molecular-orbital and Marcus-Hush theories with experiments. AB - The highly disparate rates of aromatic nitrosation and nitration, despite the very similar (electrophilic) properties of the active species: NO(+) and NO(2)(+) in Chart 1, are quantitatively reconciled. First, the thorough mappings of the potential-energy surfaces by high level (ab initio) molecular-orbital methodologies involving extensive coupled-cluster CCSD(T)/6-31G optimizations establish the intervention of two reactive intermediates in nitration (Figure 8) but only one in nitrosation (Figure 7). Second, the same distinctive topologies involving double and single potential-energy minima (Figures 6 and 5) also emerge from the semiquantitative application of the Marcus-Hush theory to the transient spectral data. Such a striking convergence from quite different theoretical approaches indicates that the molecular-orbital and Marcus-Hush (potential energy) surfaces are conceptually interchangeable. In the resultant charge transfer mechanism, the bimolecular interactions of arene donors with both NO(+) and NO(2)(+) spontaneously lead (barrierless) to pi-complexes in which electron transfer is concurrent with complexation. Such a pi-complex in nitration is rapidly converted to the sigma-complex, whereas this Wheland adduct in nitrosation merely represents a high energy (transition-state) structure. Marcus Hush analysis thus demonstrates how the strongly differentiated (arene) reactivities toward NO(+) and NO(2)(+) can actually be exploited in the quantitative development of a single coherent (electron-transfer) mechanism for both aromatic nitrosation and nitration. PMID- 12630884 TI - Optimization of tether length in nonglycosidically linked bivalent ligands that target sites 2 and 1 of a Shiga-like toxin. AB - A series of bivalent ligands for a Shiga-like toxin have been synthesized, their experimentally determined inhibitory activities were compared with a simplified thermodynamic model, and computer simulations were used to predict the optimal tether length in bivalent ligands. The design of the inhibitors exploits the proximity of the C-2' hydroxyl groups of two P(k)-trisaccharides when bound to two different, neighboring carbohydrate recognizing binding sites located on the surface of Shiga-like toxin. NMR studies of the complex between the toxin and bivalent ligands show that site 2 and site 1 of a single B subunit are simultaneously occupied by a tethered P(k)-trisaccharide dimer. A simplified thermodynamic treatment provides the intrinsic affinities and binding energies for the intermolecular and intramolecular association events and permits the deconvolution of the contributions to the relative binding energies for the set of bivalent ligands. Conformational analysis based on MD simulations for bivalent galabioside dimers containing different tethers demonstrated that the calculated local concentrations of the pendant ligand at the second binding site correlate with the experimentally determined relative affinity values of the respective bivalent ligands, thereby providing a predictive method to optimize tether length. PMID- 12630885 TI - Single crystalline commensurate metallic assemblages of pi-slabs and CdI2-type layers: synthesis and properties of beta-(EDT-TTF-I2)2[Pb5/6 square 1/6I2](3) and beta-(EDT-TTF-I2)2[Pb2/3+xAg1/3-2x square xI2]3, x = 0.05. AB - The ability of I...I van der Waals interactions to direct the self-assembly of slabs of the radical cation of ethylenedithio-1,2-diiodo-tetrathiafulvalene, EDT TTF-I(2), and polymeric lead iodide covalent anionic layers is demonstrated by the synthesis of single crystals of beta-(EDT-TTF-I(2))(2)(.+)[(Pb(5/6) square (1/6)I(2))(1/3-)](3), triclinic, P-1, a = 7.7818(8), b = 7.9760(8), c = 19.9668(2) A, alpha = 82.409(12), beta = 85.964(12), gamma = 61.621(11) degrees, V = 1080.76(19) A(3), R1, wR2 = 0.0459, 0.0947; and beta-(EDT-TTF I(2))(2)(.+)[(Pb(2/3+x)Ag(1/3-2x)square(x)I(2))(1/3-)](3), x approximately 0.05, triclinic, P-1, a = 7.7744(8), b = 7.9193(8), c = 19.834(2) A, alpha = 87.189(12), beta = 83.534(12), gamma = 61.602(11) degrees, V = 1067.4(2) A(3), R1, wR2 = 0.0508, 0.0997. The state-of-the-art, combined microprobe and structural analysis of the metal site vacancies and occupancies patterns reveal a commensurate organic-inorganic interface and point out the importance of halogen.halogen van der Waals interactions to future studies aiming at directing interface topologies. The electronic structure, room-temperature metallic character and metal-insulator transition at ca. 70 K of the two-dimensional organic slabs are retained upon alloying of the inorganic sublattice with monocations. The room-temperature conductivity of the metallic lead-silver alloy is 2 orders of magnitude larger than that of beta-(EDT-TTF-I(2))(2)(.+)[(Pb(5/6) square (1/6)I(2))(1/3-)](3). This calls for the study of materials with diverse alloying patterns with metal cations of different nature and charge. PMID- 12630886 TI - Self-assembly of fluid-filled KHCO(3) microfibers. AB - Self-assembly of KHCO(3) fibers is observed when glassy oligomerized films of poly[(aminopropyl)siloxane] containing K(+) ions, denoted K(+)/poly-APS, are exposed to CO(2) and H(2)O. The fibers are crystalline, narrow (0.4-3 microm diam), high aspect ratio (up to at least 300), and, on the basis of Raman spectroscopy, dominated by KHCO(3). The fibers contain fluid that is dominated by aqueous potassium formate (KOOCH). A multistep phenomenological model is proposed to account for the self-assembly. PMID- 12630887 TI - First X-ray characterization and theoretical study of pi-alkyne, alkynyl-hydride, and vinylidene isomers for the same transition metal fragment [Cp*Ru(PEt3)2]+. AB - The reaction of the chloro-complex [CpRuCl(PEt(3))(2)] with acetylene gas in methanol gave the pi-alkyne complex [CpRu(eta(2)-HCtbd1;CH)(PEt(3))(2)][BPh(4)] (1), which has been structurally characterized by X-ray analysis. The alkyne complex undergoes spontaneous isomerization even at low temperature, yielding the metastable alkynyl-hydride complex [CpRu(H)(Ctbd1;CH)(PEt(3))(2)][BPh(4)] (2), as the result of the oxidative addition of the alkyne C-H bond. This compound has also been structurally characterized despite it tautomerizes spontaneously into the stable primary vinylidene [CpRu(=C=CH(2))(PEt(3))(2)][BPh(4)] (3). This species has been alternatively prepared by a two-step deprotonation/protonation synthesis from the pi-alkyne complex. Moreover, the reaction of the initial chloro-complex with monosubstituted alkynes HCtbd1;CR (R = SiMe(3), Ph, COOMe, (t)Bu) has been studied without detection of pi-alkyne intermediates. Instead of this, alkynyl-hydride complexes were obtained in good yields. They also rearrange to the corresponding substituted vinylidenes. In the case of R = SiMe(3), the isomerization takes place followed by desilylation, yielding the primary vinylidene complex. X-ray crystal structures of the vinylidene complexes [CpRu(=C=CH(2))(PEt(3))(2)][BPh(4)] (3) and [CpRu(=C=CHCOOMe)(PEt(3))(2)][BPh(4)] (10) have also been determined. Both, full ab initio and quantum mechanics/molecular mechanics (QM/MM) calculations were carried out, respectively, on the model system [CpRu(C(2)H(2))(PH(3))(2)](+) (A) and the real complex [CpRu(C(2)H(2))(PEt(3))(2)](+) (B) to analyze the steric and electronic influence of ligands on the structures and relative energies of the three C(2)H(2) isomers. QM/MM calculations have been employed to evaluate the role of the steric effects of real ligands, whereas full ab initio energy calculations on the optimized QM/MM model have allowed recovering the electronic effects of ligands. Additional pure quantum mechanics calculations on [CpRu(C(2)H(2))(PH(3))(2)](+) (C) and [CpRu(C(2)H(2))(PMe(3))(2)](+) (D) model systems have been performed to analyze in more detail the effects of different ligands. Calculations have shown that the steric effects induced by the presence of bulky substituents in phosphine ligand are responsible for experimentally observed alkyne distortion and for relative destabilization of the alkyne isomer. Moreover, increasing the phosphine basicity and sigma donor capabilities of ligands causes a relative stabilization of an alkynyl-hydride isomer. The combination of both steric and electronic effects, makes alkyne and alkynyl hydride isomers to be close in energy, leading to the isolation of both complexes. PMID- 12630888 TI - Photonic crystal carbohydrate sensors: low ionic strength sugar sensing. AB - We developed a carbohydrate sensing material, which consists of a crystalline colloidal array (CCA) incorporated into a polyacrylamide hydrogel (PCCA) with pendent boronic acid groups. The embedded CCA diffracts visible light, and the PCCA diffraction wavelength reports on the hydrogel volume. This boronic acid PCCA responds to species containing vicinal cis diols such as carbohydrates. This PCCA photonic crystal sensing material responds to glucose in low ionic strength aqueous solutions by swelling and red shifting its diffraction as the glucose concentration increases. The hydrogel swelling results from a Donnan potential due to formation of boronate anion; the boronic acid pK(a) decreases upon glucose binding. This sensing material responds to glucose and other sugars at <50 microM concentrations in low ionic strength solutions. PMID- 12630889 TI - Evidence for a lack of reactivity of carotenoid addition radicals towards oxygen: a laser flash photolysis study of the reactions of carotenoids with acylperoxyl radicals in polar and non-polar solvents. AB - In this paper, we report the results of a laser flash photolysis study of the reactions of a range of carotenoids with acylperoxyl radicals in polar and nonpolar solvents. The results show, for the first time, that carotenoid addition radicals do not react with oxygen to form carotenoid peroxyl radicals; an observation which is of significance in relation to antioxidant/pro-oxidant properties of carotenoids. Acylperoxyl radicals, generated by photolysis of ketone precursors in oxygenated solvents, display high reactivity toward carotenoids in both polar and nonpolar solvents, but the nature of the carotenoid radicals formed is dependent on solvent polarity. In hexane, acylperoxyl radicals react with carotenoids with rate constants in the region of 10(9) M(-1) s(-1) and give rise to transient absorption changes in the visible region that are attributed to the formation of addition radicals. All of the carotenoids show bleaching in the region of ground-state absorption and, with the exception of 7,7'-dihydro-beta-carotene (77DH), no distinct absorption features due to addition radicals are observed beyond the ground state absorption region. For 77DH, the addition radical displays an absorption band that is spectrally resolved from the parent carotenoid absorption. The rate of decay of the 77DH addition radical is unaffected by oxygen in the concentration range 10(-4)-10(-2) M, suggesting that these resonance-stabilized carbon-centered radicals are not scavenged by oxygen. At low incident laser intensities, the 77DH addition radical decay kinetics are 1st order with k(1) approximately 4 x 10(3) s(-1) at room temperature. The 1st order decay is attributed to an intramolecular cyclization process, which is supported by the substantial negative entropies of activation obtained from measurements of the decay rate constants for different 77DH addition radicals as a function of temperature. No transient absorption features are observed in the red or near-infrared regions in hexane for any of the carotenoids studied. In polar solvents such as methanol, acylperoxyl radicals also react with carotenoids with rate constants in the region of 10(9) M(-1) s( 1), but give rise to transient absorption changes in both the visible and the red/near-infrared regions, where it is evident that there are two distinct species. For 77DH, the addition radical absorption around 450 nm is still evident, although its kinetic behavior differs from its behavior in hexane. For 77DH and zeta-carotene (zeta-CAR) the spectral and kinetic resolution of the various absorption bands simplifies kinetic analysis. The kinetic evidence suggests that addition radical formation precedes formation of the two near infrared absorbing species, and that the kinetics of the addition radical decay match the kinetics of formation of the first of these species (NIR1, absorbing at shorter wavelengths). The decay of NIR1 leads to NIR2, which is attributed to the carotenoid radical cation. The solvent dielectric constant dependence of the relative amounts of NIR1 and NIR2 formed leads us to speculate that NIR1 is an ion-pair. However, an alternative assignment for NIR1 is an isomer of the radical cation. The results, in terms of the pattern of reactivity the carotenoids display and of the properties of the carotenoid radicals formed, are discussed in relation to the antioxidant/pro-oxidant properties of carotenoids. PMID- 12630890 TI - On the dissolution processes of Na2I+ and Na3I2+ with the association of water molecules: mechanistic and energetic details. AB - The sequential association energies for one through six water molecules clustering to Na(2)I(+), as well as one and two water molecules clustering to Na(3)I(2)(+), are measured. The association energies show a pairwise behavior, indicating a symmetric association of water molecules to the linear Na(2)I(+) and Na(3)I(2)(+) ions. This pairwise behavior is well reproduced by Density Functional Theory (DFT) calculations. DFT calculations also suggest that a significant separation of charge for the Na-I ion pair occurs when four or more water molecules cluster to a single sodium center. Two different solvent separated ion pairs have been identified with the DFT calculations. Experiments also show that the dissolution processes, loss of a neutral NaI unit, occurs when six or more water molecules have been added to Na(2)I(+) cluster. However, one or two water molecules are able to detach an NaI unit from the Na(3)I(2)(+) cluster. The difference in solubility of the Na(2)I(+) and Na(3)I(2)(+) ions is due to the difference in the energies required to lose an NaI unit from these two species. The experiment also confirms that the loss of a neutral NaI unit, instead of an Na(+) ion, occurs during the dissolution processes of Na(3)I(2)(+). The microsolvation schemes proposed to explain our experimental observations are supported by DFT and phase space theory (PST) calculations. PMID- 12630891 TI - Mechanism and energetics of intramolecular hydrogen transfer in amide and peptide radicals and cation-radicals. AB - Intramolecular hydrogen transfer in five model amide and peptide radicals and cation-radicals was investigated by combined B3LYP-MP2 calculations. Hypervalent ammonium radicals produced by electron capture in protonated peptides undergo competitive elimination of ammonia, H-atom loss, and H-atom migration to neighboring amide carbonyls. The calculated transition state energies for H-atom migration are slightly but uniformly lower than those for H-atom loss. Transition state theory calculations with inclusion of quantum tunneling effects predict k(H migration)/k(H loss) branching ratios that increase with the ring size of the cyclic transition state for the migration. Intramolecular hydrogen-atom migration in amide and peptide radicals can be described by the proton-coupled electron transfer mechanism. The migrating hydrogen atom shows a negligible spin density and substantial positive charge that are typical of a proton migration. Electron transfer occurs through a pi-orbital system and proceeds in the same (clockwise) or opposite (counterclockwise) direction as the proton motion, depending on the electronic properties of the chain connecting the ammonium group and the amide bond. PMID- 12630892 TI - A route for bulk separation of semiconducting from metallic single-wall carbon nanotubes. AB - Substantial separation of single-wall carbon nanotubes (SWNTs) according to type (metallic versus semiconducting) has been achieved for HiPco and laser-ablated SWNTs. We presently argue that stable dispersions of SWNTs with octadecylamine (ODA) in tetrahydrofuran (THF) originate from the physisorption and organization of ODA along the SWNT sidewalls in addition to the originally proposed zwitterion model. Furthermore, the reported affinity of amine groups for semiconducting SWNTs, as opposed to their metallic counterparts, contributes additional stability to the physisorbed ODA. This provides a venue for the selective precipitation of metallic SWNTs upon increasing dispersion concentration, as indicated by Raman investigations. PMID- 12630893 TI - Relaxation-assisted separation of chemical sites in NMR spectroscopy of static solids. AB - We discuss the potential use of relaxation times toward the resolution of inequivalent chemical sites in the NMR spectroscopy of powdered or disordered samples. This proposal is motivated by the significant differences that can often be detected in the relaxation behavior of sites in solids, particularly when focusing on NMR observations of quadrupolar nuclei possessing different coordination and/or dynamic environments. It is shown that in these cases the implementation of a non-negative least-squares analysis on relaxation data sets enables the bidimensional resolution of overlapping powder line shapes, even when dealing with static samples. In combination with signal-enhancement methodologies such as the quadrupolar Carr-Purcell Meiboom-Gill train, such relaxation-assisted separations open up valuable routes toward the high-resolution characterization of systems involving insensitive (e.g., low-gamma) nuclei. The principles and limitations of the 2D NMR approach resulting from these considerations are discussed, and their potential is exemplified with a variety of static and spinning investigations. Their extension to other nuclear systems where spectral resolution is problematic, such as protons in organic solids, is also briefly considered. PMID- 12630894 TI - Catalytic gas phase oxidation of methanol to formaldehyde. AB - Two gas-phase catalytic cycles for the two-electron oxidation of primary and secondary alcohols were detected by multistage mass spectrometry experiments. A binuclear dimolybdate center [Mo(2)O(6)(OCHR(2))](-) acts as the catalyst in both these cycles. The first cycle proceeds via three steps: (1) reaction of [Mo(2)O(6)(OH)](-) with alcohol R(2)HCOH and elimination of water to form [Mo(2)O(6)(OCHR(2))](-); (2) oxidation of the alkoxo ligand and its elimination as aldehyde or ketone in the rate-determining step; and (3) regeneration of the catalyst via oxidation by nitromethane. Step 2 does not occur at room temperature and requires the use of collisional activation to proceed. The second cycle is similar but differs in the order of reaction with alcohol and nitromethane. The nature of each of these reactions was probed by kinetic measurements and by variation of the substrate alcohols (structure and isotope labeling). The role of the binuclear molybdenum center was assessed by examination of the relative reactivities of the mononuclear [MO(3)(OH)](-) and binuclear [M(2)O(6)(OH)](-) ions (M = Cr, Mo, W). The molybdenum and tungsten binuclear centers [M(2)O(6)(OH)](-) (M = Mo, W) were reactive toward alcohol but the chromium center [Cr(2)O(6)(OH)](-) was not. This is consistent with the expected order of basicity of the hydroxo ligand in these species. The chromium and molybdenum centers [M(2)O(6)(OCHR(2))](-) (M = Cr, Mo) oxidized the alkoxo ligand to aldehyde, while the tungsten center [W(2)O(6)(OCHR(2))](-) did not, instead preferring the non-redox elimination of alkene. This is consistent with the expected order of oxidizing power of the anions. Each of the mononuclear anions [MO(3)(OH)](-) (M = Cr, Mo, W) was inert to reaction with methanol, highlighting the importance of the second MoO(3) unit in these catalytic cycles. Only the dimolybdate center has the mix of properties that allow it to participate in each of the three steps of the two catalytic cycles. The three reactions of these cycles are equivalent to the three essential steps proposed to occur in the industrial oxidation of gaseous methanol to formaldehyde at 300-400 degrees C over solid-state catalysts based upon molybdenum(VI)-trioxide. The new gas-phase catalytic data is compared with those for the heterogeneous process. PMID- 12630897 TI - Comparison of cellular stress levels and green-fluorescent-protein expression in several Escherichia coli strains. AB - Constructs comprising stress-gene promoter elements from rpoH (Sigma 32), clpB or dnaK linked to a green-fluorescent-protein (GFP) expression vector were previously used as non-invasive "stress probes" in Escherichia coli. We compared cellular stress responses in four E. coli strains: production hosts JM105 and BL21, and cloning hosts HB101 and TOP10. When GFP was also used as a model for foreign protein production, we generally observed that the level of expression was inversely proportional to the level of cellular stress. JM105 showed the highest cellular stress level and very low GFP expression, while BL21 exhibited the lowest cellular stress level and the highest GFP expression, in both normal and heat-shock stress environments. PMID- 12630898 TI - Construction of a new tumour necrosis factor fusion-protein expression vector for high-level expression of heterologous genes in Escherichia coli. AB - We report the construction and application of a new fusion-protein expression plasmid (TNFHis) for Escherichia coli. The plasmid contains both P(R) and P(L) promoters and is optimized to allow a higher level of expression of mature coding sequences. It also contains a six-histidine tag for convenient purification as well as thrombin and hydroxylamine recognition sites for cleaving heterologous protein. The potential use of this expression vector is demonstrated by comparing the expression levels of human tumour necrosis factor (TNF), interferon, interleukin 11, colony-forming factor, osteoprotegrin and interleukin 2 in E. coli. Furthermore, all expressed TNF fusion proteins can be detected by anti-TNF alpha antibody or by specific antibodies and purified by Ni(2+)-nitrilotriacetate beads. The expressed TNF fusion proteins can be cleaved by hydroxylamine. PMID- 12630899 TI - An improved methodology to produce Flavobacterium heparinum chondroitinases, important instruments for diagnosis of diseases. AB - Chondroitinases are very important tools for the identification and structural analysis of proteoglycans. Enzymic analysis with Flavobacterium heparinum chondroitinases has shown that chondroitin sulphate and dermatan sulphate structures are modified in many human diseases, suggesting a diagnostic value for these enzymes. Furthermore, it was recently shown that F. heparinum chondroitinases AC and B inhibit tumoural cell growth, invasion and angiogenesis. Due to the increasing importance of F. heparinum chondroitinases, we investigated optimized conditions for preparation and assay of chondroitinases AC, B and C. The Dimethylmethylene Blue assay was modified and fully developed to measure the chondroitinase activities of crude extracts of F. heparinum. This method estimates chondroitin sulphate or dermatan sulphate depolymerization upon the digestion of chondroitinase, and was compared with A (232), which measures the unsaturated products formed. Trypticase was the best culture medium, both for bacterial growth and enzyme induction. The chondroitinases were solubilized by ultrasound under conditions that do not completely disrupt the cells, suggesting that they are located at the periplasmic space. Maximum chondroitinase induction occurred in the presence of 0.2-1.0 g/l chondroitin sulphate. Chondroitin sulphate-degradation products were also inducers, but heparin and heparan sulphate were not. Chondroitinases AC, B and C were separated from each other by hydrophobic-interaction chromatography on Phenyl-Sepharose HP. When contaminant proteins were first removed from crude extract by Q-Sepharose, the chondroitinases could be purified to homogeneity in this phenyl-Sepharose chromatographic step. PMID- 12630900 TI - Kinetic study of recombinant human protein disulphide isomerase-assisted C125A recombinant human interleukin-2 folding. AB - A kinetic model was developed to describe recombinant human protein disulphide isomerase (rhPDI)-assisted folding of a substrate protein, C125A recombinant human interleukin-2 (C125A rhIL-2). A series of progress curves showing native C125A rhIL-2 formation under different reaction conditions were generated. Non linear regression analysis of the progress curves of rhPDI-assisted C125A rhIL-2 folding was used to fit the differential equations of the described kinetic models. The goodness-of-fit of the model to the experimental datasets was used to support or exclude a particular kinetic model of rhPDI-assisted C125A rhIL-2 folding. The results suggest that the formation of native C125A rhIL-2 results from both glutathione-dependent oxidative folding and rhPDI-catalysed folding reactions. During oxidative folding of C125A rhIL-2, both rhPDI and reduced C125A rhIL-2 aggregated in folding buffer. The aggregation rates of rhPDI and C125A rhIL-2 followed second-order kinetics. Guanidinium chloride inactivated rhPDI but also decreased the aggregation of reduced C125A rhIL-2. These results demonstrate that during rhPDI-assisted C125A rhIL-2 folding, reduced C125A rhIL-2 aggregation competes with the productive folding pathway. While rhPDI enhances the oxidative folding of C125A rhIL-2, inactivation of rhPDI by the residual guanidinium chloride compromises its catalytic efficiency. The established model can be used to optimize the folding components in the folding mixture, and thus improve the folding efficiency. PMID- 12630901 TI - Phytoremediation of 2,4-dichlorophenol by Brassica napus hairy root cultures. AB - We have obtained hairy root cultures of Brassica napus with high biomass and genetic stability which produce peroxidases, enzymes involved in biodegradation processes. In this work, these hairy root cultures were used to study the removal of 2,4-dichlorophenol (2,4-DCP), a common contaminant in industrial effluents that is highly toxic for human and aquatic life. The optimum conditions to obtain high efficiency in the removal process were established. Roots were able to remove 2,4-DCP from aqueous solutions containing 100-1000 mg/l, in the presence of H(2)O(2) concentrations ranging from 5 to 10 mM. After a short period of incubation (15 min), high removal efficiencies were achieved (91-94%) and maximal removal, of approx. 97-98%, was obtained with 1 h of reaction. High removal efficiencies (93-95%) were observed in a broad pH range (pH 3-9), reaching 98-99% in the range pH 4-8. Moreover, roots could be re-used, almost for six consecutive cycles, to remove 2,4-DCP. The oxidation catalysed by peroxidases would be the main mechanism involved in this process. The results suggest that these cultures could be useful tools for phytoremediation. PMID- 12630902 TI - Enhanced production of verbenol, a highly valued food flavourant, by an intergeneric fusant strain of Aspergillus niger and Penicillium digitatum. AB - An intergeneric hybrid, obtained from Aspergillus niger and Penicillium digitatum, biotransformed (-)-cis-alpha-pinene (5 mg/25 ml) into (-)-cis-verbenol (60%; 1.08 mg/g of biomass) in 6 h compared with 0.18 mg/g of biomass for verbenol (10-15%; 0.18-0.27 mg/g of biomass) in the initial parent cultures. PMID- 12630903 TI - Enzymic cleavage of fusion protein using immobilized urokinase covalently conjugated to glyoxyl-agarose. AB - We immobilized urokinase (UK) by covalent attachment to activated Sepharose 6B-CL through multi-point amine coupling and evaluated its performance in cleaving a fusion protein, which consisted of recombinant human growth hormone (hGH) and a fragment of glutathione S-transferase that was linked by a tetrapeptide of a UK specific recognition sequence. Packing densities of aldehyde groups on the activated agarose surface could be controlled in a gel range of 7-60 micromol/ml aldehyde by the amount of glycidol used. The immobilization yield was nearly 100% at pH 10.5, and the specific activity of the immobilized UK was equivalent to about 80% of soluble UK under the assay conditions. The immobilized UK showed an improvement in pH and thermal stability, probably due to the structural rigidity imparted by multi-point linkages to the matrix. The cleavage rate by the immobilized UK was lower than that of the soluble enzyme but the side reaction of cryptic cleavage was significantly decreased, which might suggest that the enzyme's specificity was altered by the immobilization. Cleavage yield in the column packed with immobilized UK was dependent on the feed rate, and the yield was approx. 80% of that of the soluble UK. The monomeric hGH could be obtained by selectively precipitating the uncleaved fusion protein and the GST fragments at an acidic pH. PMID- 12630904 TI - Clostridium perfringens alpha-N-acetylgalactosaminidase blood group A2-degrading activity. AB - Enzymic modification of type A(2) erythrocyte membranes with Clostridium perfringens alpha-N-acetylgalactosaminidase was investigated. An ELISA demonstrated hydrolysis of type A(2) epitopes under conditions of red-blood-cell collection and storage. The enzyme hydrolysed the terminal N-acetyl-alpha-D galactosamine from the blood type A(2) antigen, producing H antigen, blood group O, which is universally compatible in the ABO system. The enzyme was active in common red-cell preservative solutions at pH 6.4-7.0, at 4 degrees C, at ionic strengths found in stored red cell units and in the presence of type A plasma. These data imply that the C. perfringens alpha-N-acetylgalactosaminidase might be added directly to packed A(2) red-blood-cell units for enzymic conversion to blood type O. Further studies are warranted. PMID- 12630905 TI - Regulatory effects of alanine-group amino acids on serine alkaline protease production by recombinant Bacillus licheniformis. AB - Influences of the concentration and addition time of alanine-group amino acids, i.e. alanine, leucine and valine, on serine alkaline protease (SAP) synthesis were investigated by Bacillus licheniformis (DSM 1969) carrying pHV1431:: sub C in a defined medium to identify the amino acids creating intracellular reaction rate limitation in SAP production. While the precursors of alanine-group amino acids, pyruvate and alanine, did not affect SAP production considerably within the range 0-15 mM, the addition of leucine decreased both SAP production and cell formation, because of the inhibition of valine synthesis. Although valine inhibits reactions starting with pyruvate towards 2-oxo-isovalerate, due to conversion of valine into 2-oxo-isovalerate and from 2-oxo-isovalerate to leucine, valine did not inhibit leucine synthesis. Val (7.5 mM) supply at t =0 h increased SAP activity to an activity of 1070 units.cm(-3) which was 1.3-fold higher than that of the reference production medium. The highest cell growth yield on substrate (Y (X/S)) was obtained as 0.24 g.g(-1) with the supply of alanine; and the highest product formation yield on substrate was obtained as 0.134 units.g(-1) with the supply of valine. By using the results obtained, strategies for increasing SAP production and complex medium design were also discussed. PMID- 12630906 TI - Concentration and removal of prion proteins from biological solutions. AB - The pathogenesis of prion diseases is characterized by the accumulation of amyloid-like rods or scrapie-associated fibrils. The major protein component of scrapie-associated fibrils is an abnormally folded isoform of the normal cellular prion protein (PrP(C)) that is resistant to digestion by proteinase K and is referred to as PrP(Sc). Purified human recombinant (hr PrP) was used to characterize the binding of a set of RNAs with affinity to PrP proteins. We report here that hr PrP has two RNA-binding activities at physiological pH. One activity is capable of binding all of the screened RNAs with high affinity, whereas the other activity can bind only to a subset of the RNAs with high affinity in the presence of non-specific competitor RNAs. A novel RNA belonging to the latter class, RQ11+12, bound to hr PrP with high affinity in the presence of vast molar excesses of competing RNAs. Beads impregnated with the RQ11+12 RNA were used to construct a filtration column. The column efficiently bound hr PrP and native PrP(C) from serum and urine. Importantly, the filtration device was also capable of binding proteinase K-treated PrP(Sc) from serum and urine. The level of sensitivity of detection of PrP by standard Western blotting was increased at least 1000-fold by first concentrating PrP from solution with the filtration column. PMID- 12630907 TI - Single-step purification of a protein-folding catalyst, the SlyD peptidyl prolyl isomerase (PPI), from cytoplasmic extracts of Escherichia coli. AB - The protein-folding catalyst SlyD, a peptidyl prolyl cis-trans isomerase regulated by metal binding, was initially discovered as a major contaminant of non-denaturing immobilized metal-affinity chromatography (IMAC)-based procedures for the purification of heterologously expressed 6xHis-tagged proteins from Escherichia coli. Given its ability to bind weakly to nickel-nitrilotriacetic acid (Ni(2+)-NTA), protocols for the purification of SlyD currently use an initial non-denaturing IMAC step, followed by an ion-exchange chromatographic step and occasionally also other chromatographic steps. Here we demonstrate that using denaturing conditions instead of non-denaturing conditions, and by processing of large quantities of culture through small volumes of Ni(2+)-NTA resin to increase competition for binding, single-step purification of SlyD to homogeneity can be achieved directly from E. coli extracts, as assessed through spectroscopic and electrophoretic methods. The purified, denatured SlyD is shown to be capable of refolding to give rise to a native-like CD spectrum, establishing the utility of the procedure. The procedure also establishes SlyD to be the only E. coli protein capable of contaminating denaturing IMAC-based procedures. PMID- 12630908 TI - Overexpression of the pectin lyase gene of Pseudomonas marginalis in Escherichia coli and purification of the active enzyme. AB - A pectin lyase gene (pnl) of Pseudomonas marginalis was cloned and overexpressed in Escherichia coli BL21(DE3). The pnl gene was amplified by PCR, inserted into pET29c with a six-His tag and the overproduced active enzyme was purified almost to homogeneity using a Ni(2+)-nitrilotriacetate-agarose column. The purified pectin lyase (PNL; EC 4.2.2.10, family 1) is inhibited by NAD+ (at concentrations above 0.25 mM), NADH or dithiothreitol. Evidence for the existence of a heat labile protein inhibitor of PNL is also reported. The DNA-binding ability of PNL was demonstrated by DNA-retardation experiments. The partially purified enzyme was incubated with plasmid DNA and the complex was shifted to a higher molecular mass. Analysis of the electroeluted proteins from the protein-DNA complex revealed that one of the electroeluted protein bands was PNL. Antibodies against the overexpressed PNL were also prepared and partially purified. PMID- 12630909 TI - Low-molecular-mass thiol compounds from a free-living highly pathogenic amoeba, Naegleria fowleri. AB - Acid extracts labelled with the fluorescent reagent monobromobimane and separated by HPLC have enabled the detection of low-molecular-mass thiol compounds in Naegleria fowleri for the first time. The amounts detected are expressed in nmol/1 x 10(6) trophozoites cultivated at various stages of growth in the appropriate culture medium. N. fowleri is a highly pathogenic free-living amoeba, in which we found important thiol compounds, some of them in their reduced and oxidized forms. Unlike cysteine and glutathione, a number of these are not represented in normal human lymphocytes. Some of these thiol compounds from Naegleria must have their respective disulphide reductases, although the presence of thiol-disulphide exchange reactions must be considered. Ovothiol A, with antioxidant properties, is an example of a compound that is kept reduced by trypanothione in trypanosomatids, although no disulphide reductase for ovothiol A has yet been discovered. In our case we were unable to detect this biothiol in Naegleria. The presence of thiol compounds that seem to be particular to this pathogen and which are not present in human lymphocytes opens the possibility of searching for disulphide-reducing enzymes that can serve as drug targets. PMID- 12630910 TI - Post-synaptic density-95 promotes calcium/calmodulin-dependent protein kinase II mediated Ser847 phosphorylation of neuronal nitric oxide synthase. AB - Post-synaptic density-95 (PSD-95) is a neuronal scaffolding protein that associates with N -methyl-D-aspartate (NMDA) receptors and links them to intracellular signalling molecules. In neurons, neuronal nitric oxide synthase (nNOS) binds selectively to the second PDZ domain (PDZ2) of PSD-95, thereby exhibiting physiological activation triggered via NMDA receptors. We have demonstrated previously that Ca(2+)/calmodulin-dependent protein kinase IIalpha (CaM-K IIalpha) directly phosphorylates nNOS at residue Ser(847), and can attenuate the catalytic activity of the enzyme in neuronal cells [Komeima, Hayashi, Naito and Watanabe (2000) J. Biol. Chem. 275, 28139-28143]. In the present study, we examined how CaM-K II participates in the phosphorylation by analysing the functional interaction between nNOS and PSD-95 in cells. The results showed that PSD-95 directly promotes the nNOS phosphorylation at Ser(847) induced by endogenous CaM-K II. In transfected cells, this effect of PSD-95 required its dual palmitoylation and the PDZ2 domain, but did not rely on its guanylate kinase domain. CaM-K Ialpha and CaM-K IV failed to phosphorylate nNOS at Ser(847) in transfected cells. Thus PSD-95 mediates cellular trafficking of nNOS, and may be required for the efficient phosphorylation of nNOS at Ser(847) by CaM-K II in neuronal cells. PMID- 12630911 TI - Sequence motifs of tissue inhibitor of metalloproteinases 2 (TIMP-2) determining progelatinase A (proMMP-2) binding and activation by membrane-type metalloproteinase 1 (MT1-MMP). AB - Fundamental cellular processes including angiogenesis and cell migration require a proteolytic cascade driven by interactions of membrane-type matrix metalloproteinase 1 (MT1-MMP) and progelatinase A (proMMP-2) that are dependent on the presence of tissue inhibitor of metalloproteinases 2 (TIMP-2). There are unique interactions between TIMP-2 and MT1-MMP, which we have previously defined, and here we identify TIMP-2 sequence motifs specific for proMMP-2 binding in the context of its activation by MT1-MMP. A TIMP-2 mutant encoding the C-terminal domain of TIMP-4 showed loss of proMMP-2 activation, indicating that the C terminal domain of TIMP-2 is important in establishing the trimolecular complex between MT1-MMP, TIMP-2 and proMMP-2. This was confirmed by analysis of a TIMP-4 mutant encoding the C-terminal domain of TIMP-2, which formed a trimolecular complex and promoted proMMP-2 processing to the intermediate form. Mutants encoding TIMP-4 from Cys(1) to Leu(185) and partial tail sequence of TIMP-2 showed some gain of activating capability relative to TIMP-4. The identified residues were subsequently mutated in TIMP-2 (E(192)-D(193) to I(192)-Q(193)) and this inhibitor showed a significantly reduced ability to facilitate proMMP-2 processing by MT1-MMP. Furthermore, the tail-deletion mutant Delta(186-194)TIMP-2 was completely incapable of promoting proMMP-2 activation by MT1-MMP. Thus the C terminal tail residues of TIMP-2 are important determinants for stable trimolecular complex formation between TIMP-2, proMMP-2 and MT1-MMP and play an important role in MT1-MMP-mediated processing to the intermediate and final active forms of MMP-2 at the cell surface. PMID- 12630912 TI - The soluble ectodomain of RetC634Y inhibits both the wild-type and the constitutively active Ret. AB - Substitution of Cys-634 in the extracellular domain of the Ret tyrosine kinase receptor causes its dimerization and activation of its transforming potential. To gain further insight into the molecular basis leading to Ret activation we purified a mutant protein consisting of the entire ectodomain of the Ret carrying a Cys-634-->Tyr substitution (EC-Ret(C634Y)). The protein is glycosylated, like the native one, and is biologically active. By using an in vitro cell system we show that EC-Ret(C634Y) inhibits the membrane-bound receptor Ret(C634Y), interfering with its dimerization. Furthermore, we demonstrate that EC-Ret(C634Y) competes with the wild-type Ret receptor for ligand binding. The results presented support the notion of the possible involvment of glial cell line derived neurotrophic factor (GDNF) with multiple endocrine neoplasia type 2A (MEN2A) tumours, and describe a useful tool for generating molecular mimetics directed towards specific mutations of the ret oncogene. PMID- 12630913 TI - Construction, expression and characterization of a chimaeric mammalian-plant aspartic proteinase. AB - Aspartic proteinases are a well-characterized class of proteinases. In plants, all nascent aspartic proteinases possess a 100-amino-acid, plant-specific sequence (PSS) within their C-terminal lobe, presumed to possess a targeting role in vivo. In this study, the PSS domain from the Arabidopsis thaliana aspartic proteinase was inserted into porcine pepsinogen at the identical location found in nascent plant aspartic proteinases, to create a chimaeric mammalian-plant enzyme. Based on enzymic activity, this chimaeric enzyme demonstrated increases in pH stability above 6 and temperature stability above 60 degrees C compared with commercial pepsin. Differential scanning calorimetry of the chimaeric enzyme illustrated an approx. 2 degrees C increase in denaturation temperature ( T (m)), with increases in co-operativity and similar enthalpy values. Kinetic analysis indicated an increase in K (m) and decreased k (cat) compared with pepsin, but with a catalytic efficiency similar to the monomeric plant aspartic proteinase from wheat. Using oxidized insulin B-chain, the chimaeric enzyme demonstrated more restricted substrate specificity in comparison with commercial pepsin. This study highlights the use of a chimaeric enzyme strategy in order to characterize unique protein domains within enzyme families, and, for the first time, a putative structure-function role for the PSS as it pertains to plant aspartic proteinases. PMID- 12630915 TI - Quantification of T cell receptor rearrangement excision circles to estimate thymic function: an important new tool for endocrine-immune physiology. AB - Although the thymus constitutes a target organ for most protein and steroid hormones, it has been quite difficult to determine the precise control exerted in vivo by the endocrine system upon thymic function. The biological role of the thymus is to ensure the generation of a diversified population of peripheral T cells able to respond to non-self-antigens but nevertheless tolerant to self antigens. For a long time, thymic function could not be monitored, as a consequence of the absence of adequate technology to differentiate recent thymic emigrants from naive T cells. The generation of T cell receptor (TCR) diversity occurs in the thymus through recombination of gene segments encoding the variable parts of the TCR alpha and beta chains. During these processes, by-products of the rearrangements are generated in the form of TCR excision circles (TRECs). As these molecules are lost upon further cell division, their quantification is actually considered as a very valuable tool to estimate thymic function. The most appropriate TREC is deltaRec-Psi(J)alpha TREC or signal joint TREC resulting from deltaRec-Psi(J)alpha rearrangement (TCRD deletion) that occurs late during thymopoiesis, before V(alpha)-J(alpha) rearrangement. Here we describe how TREC quantification is a powerful and reliable method to evaluate the impact of hormones and endocrine disorders upon thymic function. PMID- 12630914 TI - The hypothalamic-pituitary-gonadal axis: immune function and autoimmunity. AB - GnRH and sex steroids play an important role in immune system modulation and development. GnRH and the GnRH receptor are produced locally by immune cells, suggesting an autocrine role for GnRH. Experimental studies show a stimulatory action of exogenous GnRH on the immune response. The immune actions of GnRH in vivo are, however, less well established. Oestrogen and androgen receptors are expressed in primary lymphoid organs and peripheral immune cells. Experimental data have established that oestrogens enhance the humoral immune response and may have an activating role in autoimmune disorders. Testosterone enhances suppressor T cell activity. Although there are some clinical studies consistent with these findings, the impact of sex steroids in autoimmune disease pathogenesis and the risk or benefits of their usage in normal and autoimmune-disordered patients remain to be elucidated. There are neither experimental nor clinical data evaluating functional GnRH-sex steroid interactions within the human immune system, and there is a paucity of data relating to GnRH analogues, hormone replacement therapy and oral contraceptive and androgen action in autoimmune diseases. However, a growing body of experimental evidence suggests that an extra pituitary GnRH immune mechanism plays a role in the programming of the immune system. The implications of these findings in understanding immune function are discussed. PMID- 12630916 TI - Developmental changes in plasma leptin and hypothalamic leptin receptor expression in the rat: peripubertal changes and the emergence of sex differences. AB - Leptin, the peptide hormone product of the ob gene, regulates food intake and energy expenditure at the hypothalamic level via the long-form of the leptin receptor (Ob-Rb). Leptin also plays a key role in determining the onset of puberty, but there is controversy as to whether leptin provides a trigger for puberty or is a permissive signal. Thus, although leptin administration can advance puberty onset in rodents, circulating leptin appears stable across puberty. While these data suggest a permissive role for leptin in rat puberty, it is possible that a change in hypothalamic response to leptin (e.g. via increased Ob-Rb expression) could enhance leptin action and thus trigger puberty without a rise in circulating leptin. In the present study we assessed developmental changes in hypothalamic Ob-Rb mRNA and protein expression in female and male rats from late fetal to postpubertal life. Quantitative RT-PCR showed that Ob-Rb mRNA increased (P<0.05) by around fivefold from fetal to postpubertal life in both females and males. These increases in Ob-Rb mRNA expression were gradual, but did not increase significantly between postnatal day 30 (pre-puberty) and day 51 (post-puberty). By day 51, hypothalamic Ob-Rb mRNA expression was higher (P<0.05) in females relative to males. Hypothalamic Ob-Rb protein showed a comparable developmental pattern (approximate threefold increase from fetal to postpubertal life), although a significant increase (15%; P<0.05) was observed between days 30 and 51 in females. Plasma leptin levels exhibited a dynamic pattern in both male and female rats during the prepubertal period, characterised by a precipitous fall after birth, relative stability to day 5, then a rapid increase to a transient peak on day 12. Plasma leptin then remained unchanged from day 15 in female rats but increased in males after puberty, thus confirming the well recognised sex difference in adult rat leptin levels. In conclusion, this study shows that developmental increases occur not only in plasma leptin but also in hypothalamic Ob-Rb expression, suggesting that both are likely to influence the timing of puberty onset. Moreover, our data show that sex differences in both hypothalamic Ob-Rb and plasma leptin emerge only after puberty. PMID- 12630917 TI - Hypothyroidism alters mitochondrial morphology and induces release of apoptogenic proteins during rat cerebellar development. AB - Thyroid hormone (TH) deficiency leads to extensive apoptosis during cerebellar development, but the mechanism still remains unclear. Different signals also converge on mitochondria during apoptosis to induce the release of apoptogenic proteins that activate proteolytic cascade through specific enzymes called caspases. Here we studied the effect of hypothyroidism on alterations in mitochondrial structure and translocation of apoptogenic molecules during rat cerebellar development. Structural analysis of mitochondria was studied by electron microscopy. The translocation of apoptogenic molecules was analyzed by Western blotting. TH deficiency led to vacuolization, enlargement and decrease in the number of cristae. The majority of the proapoptotic molecule, Bax, was localized in mitochondria under hypothyroid conditions whereas a limited presence of Bax was detected in the euthyroid state. Translocation of cytochrome c, apoptosis-inducing factor (AIF) and second mitochondrial-derived activator of caspases (SMAC) from mitochondria to cytosol was detected primarily in early developmental stages in the hypothyroid condition. These experimental results demonstrate that TH maintains mitochondrial architecture and inhibits the release of apoptogenic molecules to prevent excess apoptosis during cerebellar development. PMID- 12630918 TI - Growth retardation induced by dexamethasone is associated with increased apoptosis of the growth plate chondrocytes. AB - Glucocorticoids cause significant growth retardation in mammals and humans and decreased proliferation of chondrocytes has been considered as the main local mechanism. Death by apoptosis is an important regulator of homeostasis in multicellular organisms. Here we chose to study the role of apoptosis in growth retardation caused by glucocorticoid treatment. We treated 7-week-old male rats with dexamethasone (5 mg/kg/day) for 7 days. Apoptosis was studied in tibiae growth plates by the TUNEL method. Immunoreactivity for parathyroid hormone related peptide (PTHrP), caspase-3, and the anti-apoptotic proteins Bcl-2 and Bcl x was also studied. Apoptosis was mainly localized in terminal hypertropic chondrocytes (THCs) in both control and dexamethasone-treated animals. Dexamethasone caused an increase in apoptosis which was fourfold in THCs (2.45+/ 0.12 vs 0.62+/-0.09 apoptotic cells/mm growth plate, P<0.001), and 18-fold in proliferative chondrocytes (0.18+/-0.04 vs 0.01+/-0.007 apoptotic cells/mm growth plate, P<0.001). Increased apoptosis after dexamethasone treatment was accompanied by increased immunoreactivity for caspase-3 and decreased immunoreactivity for the anti-apoptotic proteins Bcl-2 and Bcl-x, which further supports our apoptosis results. Dexamethasone also decreased the immunoreactivity for PTHrP, suggesting a role in the mechanism by which glucocorticoids induce apoptosis in the growth plate. We conclude that apoptosis is one mechanism involved in growth retardation induced by glucocorticoids. Premature loss of resting/proliferative chondrocytes by apoptosis could contribute to incomplete catch-up seen after prolonged glucocorticoid treatment. PMID- 12630919 TI - Vitamin K stimulates osteoblastogenesis and inhibits osteoclastogenesis in human bone marrow cell culture. AB - Accumulating evidence indicates that menaquinone-4 (MK-4), a vitamin K(2) with four isoprene units, inhibits osteoclastogenesis in murine bone marrow culture, but the reason for this inhibition is not yet clear, especially in human bone marrow culture. To clarify the inhibitory mechanism, we investigated the differentiation of colony-forming-unit fibroblasts (CFU-Fs) and osteoclasts in human bone marrow culture, to learn whether the enhancement of the differentiation of CFU-Fs from progenitor cells might relate to inhibition of osteoclast formation. Human bone marrow cells were grown in alpha-minimal essential medium with horse serum in the presence of MK-4 until adherent cells formed colonies (CFU-Fs). Colonies that stained positive for alkaline phosphatase activity (CFU-F/ALP(+)) were considered to have osteogenic potential. MK-4 stimulated the number of CFU-F/ALP(+) colonies in the presence or absence of dexamethasone. The stimulation was also seen in vitamin K(1) treatment. These cells had the ability to mineralize in the presence of alpha-glycerophosphate. In contrast, both MK-4 and vitamin K(1) inhibited 1,25 dihydroxyvitamin D(3)-induced osteoclast formation and increased stromal cell formation in human bone marrow culture. These stromal cells expressed ALP and Cbfa1. Moreover, both types of vitamin K treatment decreased the expression of receptor activator of nuclear factor kappaB ligand/osteoclast differentiation factor (RANKL/ODF) and enhanced the expression of osteoprotegerin/osteoclast inhibitory factor (OPG/OCIF) in the stromal cells. The effective concentrations were 1.0 microM and 10 microM for the expression of RANKL/ODF and OPG/OCIF respectively. Vitamin K might stimulate osteoblastogenesis in bone marrow cells, regulating osteoclastogenesis through the expression of RANKL/ODF more than through that of OPG/OCIF. PMID- 12630920 TI - Mutual antagonism of estrogen receptors alpha and beta and their preferred interactions with steroid receptor coactivators in human osteoblastic cell lines. AB - Estrogen is a major sex steroid that affects the growth, maintenance, and homeostasis of the skeleton. Two isoforms of the estrogen receptor (ERalpha and ERbeta) mediate the transcriptional effects of estrogen. Although both isoforms of ER are present and functional in some human osteoblast (OB) cell lines, there is minimal information on the differential regulation of transcription by ERalpha and ERbeta homo- or heterodimers. This report demonstrates that ERalpha and ERbeta coexpression decreases the transcriptional capacity (relative to each ER isoform alone) on an estrogen response element-dependent reporter gene in OBs but not in other non-osteoblastic cell lines. These data suggest that ERalpha and ERbeta coexpression can differentially influence the degree of transcriptional activation in certain cell types. Interestingly, the overexpression of the steroid hormone receptor coactivator-1 (SRC1) resulted in preferential transcriptional enhancement by ERbeta as well as coexpressed ERalpha and ERbeta, whereas SRC2 overexpression appeared to preferentially enhance ERalpha transactivation. SRC3 overexpression failed to enhance estrogen-dependent transcription of any ER combination in OBs. Similar overexpression experiments in COS7 cells exhibited preferential enhancement of ERalpha function with all SRCs, including SRC3. Our data also demonstrated that SRC3 mRNA is reduced in osteoblastic cells, suggesting that SRC3 may have only a minor role in these cells. These data suggest that the transactivation capacity of various ER isoforms is both SRC species and cell type dependent. PMID- 12630921 TI - Effects of the Japanese herbal medicine Keishi-bukuryo-gan and 17beta-estradiol on calcitonin gene-related peptide-induced elevation of skin temperature in ovariectomized rats. AB - The effects of a Japanese herbal medicine, Keishi-bukuryo-gan, and 17beta estradiol on calcitonin gene-related peptide (CGRP)-induced elevation of skin temperature were investigated in ovariectomized (OVX) rats. Ovariectomy not only potentiated CGRP-induced elevation of skin temperature and arterial vasorelaxation but also induced a lower concentration of endogenous CGRP in plasma and up-regulation of arterial CGRP receptors, suggesting that lowered CGRP in plasma due to ovarian hormone deficiency increases the number of CGRP receptors and consequently amplifies the stimulatory effects of CGRP to elevate skin temperature. Oral Keishi-bukuryo-gan (100-1000 mg/kg, once a day for 7 days) restored a series of CGRP-related responses observed in OVX rats by normalizing plasma CGRP levels in a dose-dependent manner as effectively as s.c. injection. 17Beta-estradiol (0.010 mg/kg, once a day for 7 days). However, Keishi-bukuryo gan did not affect the lower concentration of plasma estradiol and the decreased uterine weight due to ovariectomy, although the hormone replacement of 17beta estradiol restored them. These results suggest that Keishi-bukuryo-gan, which does not confer estrogen activity on plasma, may be useful for the treatment of hot flashes in patients for whom estrogen replacement therapy is contraindicated, as well as menopausal women. PMID- 12630922 TI - Dose- and time-dependent effects of 17beta-oestradiol on insulin sensitivity in insulin-dependent tissues of rat: implications of IRS-1. AB - Numerous studies have suggested that ovarian hormones are able to modulate insulin sensitivity, but their exact role remains unclear. We have investigated whether different doses of 17beta-oestradiol mediate changes in insulin sensitivity and if these changes could be related to modifications of insulin receptor substrate-1 (IRS-1). Female rats were ovariectomized and later separated into three groups: untreated; treated with a dose of 17beta-oestradiol sufficient to reproduce gestational plasma concentrations of 17beta-oestradiol (group E); and treated with a dose 100 times greater than that given to group E (group E2). A euglycaemic-hyperinsulinaemic clamp was used to measure insulin sensitivity. Changes in IRS-1 were analysed by Western blotting and RT-PCR assays. In group E we found a decrease in insulin sensitivity between days 11 and 16 of treatment as in late gestation, whereas in the untreated group and group E2, development of insulin resistance was observed throughout the treatment. In contrast, whereas in group E2 insulin resistance throughout the hormonal treatment was related to diminished expression and phosphorylation of IRS-1, in group E the decrease in insulin sensitivity between days 11 and 16 of treatment was not related to a decrease in IRS-1 expression. Our results suggest that the effects of oestradiol on insulin sensitivity were dose-dependent and that the insulin resistance associated with a high dose of 17beta-oestradiol was related to downregulation of IRS-1 expression. PMID- 12630923 TI - Expression of a novel factor, short-type PB-cadherin, in Sertoli cells and spermatogenic stem cells of the neonatal rat testis. AB - In the rodent testis, contact-mediated interactions between gonocytes, or neonatal stem cells, and Sertoli cells are critical for development. Previously, we showed that the neural cell adhesion molecule (NCAM) serves as a Sertoli cell gonocyte attachment factor in neonates. Its expression decreases dramatically by 1 week of age and eventually disappears in vivo, and appears to be down-regulated by thyroid hormone (tri-iodothyronine (T(3))). In this study, we used a cDNA microarray to screen for additional adhesion factors which might be important in testes of developing rats and detected expression of a novel factor, short-type PB-cadherin (STPB-C). Next, RT-PCR was used to generate cDNA for STPB-C from total RNA isolated from co-cultures, cDNA was cloned into pPCR-Script Amp SK(+) cloning vector, and plasmid DNA was isolated and sequenced to confirm the fidelity of the STPB-C cDNA portion of the plasmid. In situ hybridization analyses of testicular sections indicated that STPB-C expression in neonates is localized in the cytoplasm of many, but not all, gonocytes and in the cytoplasm of most of the surrounding Sertoli cells. Parallel hybridizations carried out on co-cultures also demonstrated a strong cytoplasmic signal in some gonocytes and in the great majority of the Sertoli cells of the underlying monolayer. With Northern analyses we found that STPB-C is expressed in vivo at high levels between days 1 and 5, with a subsequent large drop by day 10 and thereafter, suggesting that its expression may be associated with Sertoli or germ cell differentiation. Subsequent analyses of co-cultures exposed under a variety of conditions to T(3) suggest that, unlike NCAM, STPB-C is not regulated by this hormone. Next, we studied production of STPB-C protein by using an antiserum recognizing a peptide sequence unique to this factor in Western blotting and in immunolocalization. Signal was detected both intracellularly and at cell surfaces in most Sertoli cells and many gonocytes, although many of the latter cell type were also found to be negative for the protein, suggesting a potential role for STPB-C in survival and further development of some of these germ cells from which all subsequent spermatogenic cells originate. PMID- 12630924 TI - Differential regulation of rat testicular 5alpha-reductase type 1 and 2 isoforms by testosterone and FSH. AB - Testosterone is metabolised to the more potent androgen, dihydrotestosterone, by the 5alpha-reductase (5alphaR) enzyme. We previously showed that 5alpha-reduced androgens are important for maintaining androgen action on rat spermatogenesis when testicular testosterone concentrations are reduced. This study investigated expression and activity of the 5alphaR isoforms, type 1 (5alphaR-1) and type 2 (5alphaR-2), in the rat during hormone manipulation in order to understand the factors that regulate the testicular concentration of 5alphaR and testicular 5alpha-reduced androgen biosynthesis. Testicular 5alphaR-1 and 5alphaR-2 mRNA and enzyme activity were measured by real-time PCR and specific enzyme assays respectively. Hormone levels were first suppressed using two models of gonadotrophin suppression: testosterone and oestradiol treatment (LH/testosterone deficiency) or GnRH immunisation (LH/testosterone and FSH deficiency). Hormones were then either restored or suppressed for 6 days by a variety of hormonal treatments. 5alphaR-1 mRNA and enzyme activity increased when testosterone was suppressed, yet restoration of testosterone decreased 5alphaR-1 mRNA and enzyme activity, suggesting that testosterone negatively regulates 5alphaR-1. suppression of FSH decreased 5alphaR-1 mRNA yet FSH administration increased 5alphaR-1 mRNA, but no changes in 5alphaR-1 activity were observed within the 6 day period. In contrast to 5alphaR-1, testosterone did not affect the testicular concentration of 5alphaR-2 mRNA or activity, but there was evidence for modulation of 5alphaR-2 activity by FSH. Measurement of testicular androgens revealed that 5alphaR-1 was primarily responsible for the production of 5alpha reduced metabolites. It is concluded that the 5alphaR isoforms in rat testis are differentially regulated by testosterone and FSH: testosterone negatively regulated 5alphaR-1 mRNA and enzyme activity but had no affect on 5alphaR-2, whereas FSH positively regulated 5alphaR-1 mRNA and appeared to regulate 5alphaR 2. PMID- 12630925 TI - Cloning of a proopiomelanocortin cDNA from the pituitary gland of the sea bass (Dicentrarchus labrax) and assessment of mRNA expression in different tissues by means of real-time PCR. AB - Proopiomelanocortin (POMC) cDNA was cloned from sea bass (Dicentrarchus labrax) pituitary gland. A 743 nucleotide sequence was obtained coding for the following sequences flanked by sets of proteolytic cleavage sites: ACTH (Ser(88)-Met(127)), alpha-MSH (Ser(88)-Gly(102)), CLIP (Pro(106)-Met(127)), beta-LPH (Glu(131) Gln(208)), gamma-LPH (Glu(131)-Ser(175)), beta-MSH (Asp(159)-Ser(175)), and beta endorphin (Tyr(178)-Gln(208)). No region homologous to gamma-MSH/joining peptide (a tetrapod POMC feature) was found. Amino acid sequence identity was high with other teleostean species considered (tilapia: 73%) and lower with elasmobranchs (dogfish: 42%). However, the presumed biologically active peptides were highly conserved within all species considered: alpha-MSH (93-100%), ACTH (80-95%) and beta-endorphin (54-90%). Real-time PCR allowed us to quantify the expression of the POMC in different tIssues of the sea bass: pituitary gland, liver, gonad and head kidney. No significant POMC expression was found in the integument. In pituitary gland, gonads, head kidney and liver, POMC expression was respectively, 1.26x10(10), 2.67x10(5), 2.06x10(4) and 1.67x10(4) copies/ micro g mRNA. PMID- 12630926 TI - Amidated fish ghrelin: purification, cDNA cloning in the Japanese eel and its biological activity. AB - We purified ghrelin from stomach extracts of a teleost fish, the Japanese eel (Anguilla japonica) and found that it contained an amide structure at the C terminal end. Two molecular forms of ghrelin with 21 amino acids were identified by cDNA and mass spectrometric analyses: eel ghrelin-21, GSS(O-n octanoyl)FLSPSQRPQGKDKKPP RV-amide and eel ghrelin-21-C10, GSS(O-n-decanoyl) FLSPSQRPQGKDKKPPRV-amide. Northern blot and RT-PCR analyses revealed high gene expression in the stomach. Low levels of expression were found only in the brain, intestines, kidney and head kidney by RT-PCR analysis. Eel ghrelin-21 increased plasma growth hormone (GH) concentrations in rats after intravenous injection; the potency was similar to that of rat ghrelin. We also examined the effect of eel ghrelin on the secretion of GH and prolactin (PRL) from organ-cultured tilapia pituitary. Eel ghrelin-21 at a dose of 0.1 nM stimulated the release of GH and PRL, indicating that ghrelin acts directly on the pituitary. The present study revealed that ghrelin is present in fish stomach and has the ability to stimulate the secretion of GH from fish pituitary. A novel regulatory pathway of GH secretion by gastric ghrelin seems to be conserved from fish to human. PMID- 12630927 TI - Prolonged underfeeding of sheep increases myostatin and myogenic regulatory factor Myf-5 in skeletal muscle while IGF-I and myogenin are repressed. AB - The IGF axis is nutritionally sensitive in vivo and IGFs stimulate myoblast proliferation and differentiation in vitro, while myostatin inhibits these processes in vitro. We hypothesised that underfeeding would reversibly inhibit the myogenic activity of satellite cells in vivo together with decreased IGF-I and increased myostatin in muscle. Satellite cell activity was measured indirectly from the expression of proliferating cell nuclear antigen (PCNA) and the myogenic regulatory factors (MRFs), MyoD, Myf-5 and myogenin. Young sheep were underfed (30% of maintenance) and some killed after 1, 4, 12, 17, 21 and 22 weeks. Remaining underfed animals were then re-fed a control ration of pellets and killed after 2 days, and 1, 6 and 30 weeks. Expression of PCNA and MRFs decreased during the first week of underfeeding. This coincided with reduced IGF I and myostatin mRNA, and processed myostatin. Subsequently, Myf-5, MyoD, myostatin mRNA and processed myostatin increased, suggesting that satellite cells may have become progressively quiescent. Long-term underfeeding caused muscle necrosis in some animals and IGF-I and MRF expression was increased in these, indicating the activation of satellite cells for muscle repair. Re-feeding initiated rapid muscle growth and increased expression of PCNA, IGF-I and the MRFs concurrently with decreased myostatin proteins. In conclusion, these data indicate that IGF-I and myostatin may work in a coordinated manner to regulate the proliferation, differentiation and quiescence of satellite cells in vivo. PMID- 12630928 TI - Diabetes UK Annual Professional Meeting. 19-21 March 2003, Glasgow, United Kingdom. Abstracts. PMID- 12630931 TI - Surrogate measures of atherosclerosis and implications for evaluating cardiovascular risk. PMID- 12630932 TI - Sounding board: diabetes mellitus in the elderly: a truly heterogeneous entity? PMID- 12630933 TI - Measurement of metformin concentration in erythrocytes: clinical implications. AB - AIMS: Although pharmacokinetics studies have long suggested a deep compartment for the antidiabetic drug metformin, there is still little information concerning metformin accumulation by individual tissues in man. In the present study, the erythrocyte was chosen to represent this putative deep compartment and metformin concentration in erythrocytes (EM) was compared with that in plasma (PM) to delineate clinical implications. METHODS: A reference group of 58 patients with well-tolerated metformin treatment was studied to provide standard mean metformin concentrations in the fasting state. Secondly, to provide transverse data reflecting clinical practice, the authors reviewed an investigation group of 93 metformin-treated patients with available PM and EM, which had been requested either to adjust metformin dosage to renal function, or to screen for potential metformin accumulation following renal failure, metformin overdose or lactic acidosis. Thirdly, the case of an individual with major metformin accumulation was studied to provide information about metformin elimination. RESULTS: From the bulk of data, we performed three types of analyses: (1) PM and EM were compared. In the investigation group, this comparison was extended to subgroups separated according to low-to-normal, moderately increased or highly increased metformin concentration. (2) Correlative analyses of PM, EM and serum creatinine were performed. (3) A kinetic study of the spontaneous decline of PM and EM was conducted. PM and EM were, respectively, 0.5 +/- 0.4 mg/l and 0.8 +/- 0.4 mg/l in the reference group, and 11.7 +/- 17.8 mg/l (mean +/- SD, range 0.0-71.9 mg/l) and 7.5 +/- 9.4 mg/l (0.0-34 mg/l) in the investigation group, mean serum creatinine of which was 290 +/- 258 micro mol/l. In the low-to-normal PM subgroup (n = 28), PM and EM were, respectively, 0.39 +/- 0.38 mg/l and 0.84 +/- 0.68 mg/l (p < 0.001). In the moderately increased PM subgroup (from therapeutic concentrations +2 SD to 5 mg/l, n = 24), PM and EM were 2.82 +/- 1.13 mg/l and 2.72 +/- 2.03 mg/l (NS). In the sharply increased PM subgroup (> 5 mg/l, n = 41), PM and EM were 27.6 +/- 23.2 mg/l and 17.0 +/- 11.4 mg/l (p = < 0.001). PM and EM were tightly correlated (r = 0.72 in the reference group and r = 0.90 in the investigation group, p < 0.001 for both). Metformin concentrations were also correlated with those of serum creatinine, but more so in the investigation group; in subgroups, a positive correlation was found only at high metformin concentrations and in erythrocytes. The kinetic study performed in the patient with major metformin accumulation showed that PM and EM dropped within less than 3 days from a maximum concentration of 80.0 mg/l and 20.4 mg/l, respectively, to 0.67 mg/l and 6.52 mg/l. CONCLUSIONS: In conclusion, metformin appears to accumulate in erythrocytes and, consequently, may be part of a deep compartment for the drug. This evidence of slow decline in erythrocyte metformin concentration may contribute to retrospective diagnosis of metformin accumulation and to refinements in adjusting metformin dosage to renal function. PMID- 12630934 TI - A longitudinal magnetic resonance imaging (MRI) study of differences in abdominal fat distribution between normal mice, and lean overexpressers of mitochondrial uncoupling protein-3 (UCP-3). AB - AIM: To characterize evolution and distribution of abdominal adipose fat between 6 and 18 weeks of age in an animal model of energy consumption based on mice overexpressing the mitochondrial uncoupler protein 3 (UCP-3). METHODS: T2 weighted multislice MRI was performed six times during the 12 week study; visceral, subcutaneous and intermuscular fat depots were quantified. RESULTS: The overexpressor (UCP-3tg) mice consistently have less subcutaneous, visceral, interskeletal muscle and total fat throughout the experiment. Mean (standard error) volumes (ml) of the three distinct depots change between week 6 and week 18 as follows: wild type: subcutaneous 1.93 (0.28) to 6.18 (0.47), visceral 2.15 (0.34) to 6.37 (0.64), intermuscular 0.23 (0.04) to 0.53 (0.03); UCP-3tg: subcutaneous 1.47 (0.17) to 4.07 (0.57), visceral 1.18 (0.04) to 3.69 (0.59), intermuscular 0.23 (0.01) to 0.32 (0.04). Although they eat more (4.3 g compared with 3.4 g per day) the UCP-3tg's always weigh less than controls. In wild-type control animals, increases of all fat pools between week 6 and week 18 is highly significant, as it is for subcutaneous, visceral and total pools in the UCP-3tg animals. The UCP-3tg mice, however, show no significant absolute or relative increase in intermuscular fat; UCP-3 is predominantly overexpressed in skeletal muscle. CONCLUSION: MRI provides an excellent approach to comparative studies of fat distribution in animal models of energy expenditure such as the UCP-3tg mouse. PMID- 12630936 TI - Establishment of mouse as an animal model for study of diabetic cataracts: biochemical studies. AB - AIM: The primary aim of this study was to understand the pathogenesis of diabetic cataracts at biochemical level in an animal model where lens aldose reductase (AR) activity is low, similar to that in the human lens. METHODS: Mouse, which is known to have low lens AR, was selected for these studies. Diabetes was induced by intraperitoneal administration of streptozotocin. Biochemical changes in the lens were monitored in freshly isolated lenses with standard chromatographic, enzymatic and culture experiments described in the section on methods. RESULTS: The present studies provide evidence of significant biochemical changes associated with such cataract formation despite very low levels of aldose reductase. The level of glycated proteins increased to 9 mg/100 mg of total water soluble lens protein in the diabetic lenses, as compared with the normal lenses where it was only about 1.3 mg/100 mg of total protein. Glutathione (GSH), the major antioxidant in the lens, decreased from 2.35 micro mol/g in the normal lenses to about 1.17 micro mol/g in the diabetic lenses. Malonadehyde, a product of lipid peroxidation, increased from 50 micro mol/100 g in the normal to 70 micro mol/100 g in the diabetic lens. The level of adenosine triphosphate (ATP), an indicator of the overall metabolic status of the tissue, also decreased from 962 +/- 154 nmol/g in the normal to 487 +/- 130 nmol/g in the diabetic lenses. The function of the Na+-K+ ATPase was also adversely affected in diabetes, as indicated by the ability of the lens to accumulate (86)rubidium ions against its concentration gradient. The transport activity, expressed as CL/CM, was 24 in the normal lens, whereas it was only 12 in the diabetic lens. The level of sorbitol in the diabetic lens was only in the micromolar region. Hence, it was considered osmotically insignificant. CONCLUSION: Overall, the results suggest that induction of cataracts in diabetes can be related to multiple biochemical effects such as oxidative stress and glycation. Sorbitol accumulation in low aldose reductase situations, being minor, could, however, act synergistically with other factors. PMID- 12630935 TI - Socio-demographic and psychosocial factors are associated with features of the metabolic syndrome. The Women's Health in the Lund Area (WHILA) study. AB - AIM: The aim was to analyse any associations between socio-demographic and psychosocial factors and different features of the metabolic syndrome in a geographically well-defined population of middle-aged women. METHODS: A population of 10 766 Caucasian women aged 50-59 years was investigated regarding biological and socio-demographic conditions, physical activity, dietary habits, aspects of quality of life, and subjective physical and mental symptoms. The screening instrument was used to discriminate subjects as positive or negative on one or more of a total of eight variables considered to be linked to the metabolic syndrome. The cut-off values for positive screening were non-fasting capillary blood glucose >/= 8.0 mmol/l and serum triglycerides >/= 2.3 mmo/l, BMI >/= 30 kg/m2, WHR >/= 0.90, blood pressure >/= 160 and/or 95 mmHg, a family history of diabetes, and pharmacological treatment for hypertension or hyperlipidaemia. RESULTS: Altogether 6805 women (63.2%) participated: 3535 with positive and 3270 with negative screening. Multiple logistic regression analyses showed that comprehensive (OR 1.62, 95% CI 1.41-1.87) and upper secondary (1.40, 1.24-1.57) school, low physical quality of life (1.41, 1.23-1.61) and high sum of subjective physical symptoms (1.06, 1.04-1.08) were positively associated with one or more features of the metabolic syndrome, while high leisure-time exercise and healthy diet (0.84, 0.71-0.99), and low ( ID --> DD genotypes were significant as assessed by an analysis of variance. The ratio of type I:II fibers also differed according to the ACE genotype. A multivariate logistic regression analysis showed that the ACE I allele had significant additive and recessive (codominant) effects on the increased type I fibers and the ratio of type I:II fibers. No specific pattern of capillarization was observed among the three ACE genotypes. In conclusion, the ACE-I allele was associated with increased type I SMF, which may be a mechanism for the association between the ACE genotype and endurance performance. PMID- 12630963 TI - Rapid detection methods for five HGO gene mutations causing alkaptonuria. AB - Alkaptonuria (AKU) is an autosomal recessive disorder caused by the deficiency of homogentisate 1,2 dioxygenase (HGO) activity. The disease is characterized by homogentisic aciduria, ochronosis and ochronotic arthritis. AKU shows a very low prevalence (1:250 000), in most ethnic groups. Altogether 43 HGO mutations have been identified in approximately 100 patients. In Slovakia, however, the incidence of this disorder rises up to 1:19 000, and 10 different AKU mutations have been identified in this relatively small country. Here, we report detection methods developed for rapid identification of five HGO mutations. PCR primers were designed enabling detection of mutations IVS5 + 1G-->A, R58fs, and V300G by restriction digestion of amplification-created restriction sites (ACRS). Mutation G152fs is readily identified by heteroduplex analysis, and G161R by amplification refractory mutation system (ARMS) PCR. PMID- 12630964 TI - USH1C: a rare cause of USH1 in a non-Acadian population and a founder effect of the Acadian allele. AB - Usher syndrome (USH) is characterized by the associated findings of hearing loss and retinitis pigmentosa (RP), leading to progressive loss of vision. Three forms of USH can be distinguished clinically. In the most severe form, USH1, profound congenital deafness is associated with vestibular dysfunction and RP. To determine the frequency of USH1C mutations as a cause for USH1, 128 probands with Usher syndrome type 1 including seven from Acadian and 121 from non-Acadian populations were systematically screened for mutations in USH1C using a combined single-strand conformational polymorphisms (SSCP)/heteroduplex and sequencing method. All seven Acadian USH1 patients were found to be homozygous for both the 216G>A mutation and the 9-repeat VNTR which characterizes the Acadian allele, confirming previous evidence for a founder effect by haplotype analysis. However, USH1C mutations were identified in only two non-Acadian USH1 probands (1.65%) including one from Pakistan who was homozygous for a 238-239insC mutation and one from Canada was also homozygous for the Acadian allele. The low prevalence of USH1C mutations in the present study suggests that the high prevalence of the 238 239insC in Germany may reflect a founder effect. Comparison of the affected haplotypes in the Canadian patient with the Acadian USH1 patients yielded evidence for a founder effect. Our data suggest that USH1C is a relatively rare form of USH1 in non-Acadian populations and that in addition to the 216G>A Acadian mutation, the 238-239insC mutation appears to be common in some populations. PMID- 12630966 TI - CARD15/NOD2 mutational analysis in Japanese patients with Crohn's disease. PMID- 12630965 TI - Mutation analysis of Connexin 31 (GJB3) in sporadic non-syndromic hearing impairment. AB - Mutations in GJB3, the gene encoding the gap junction protein Connexin 31 (CX31), have been pathogenically linked to erythrokeratodermia and non-syndromic autosomal dominant (DFNA2) or recessive hereditary hearing impairment (HHI). To determine the contribution of CX31 to sporadic deafness, we assessed 63 individuals with non-syndromic hearing impairment for CX31 mutations. Single coding exon of CX31 was amplified from genomic DNA and then sequenced. Single nucleotide sequence alteration was present in 15 out of 63 patients (24%), all of the positives being heterozygous for the four different single base pair changes that were detected: C94T, C201T, C357T and C798T. Of these, only C94T transition, identified in two patients, results in amino acid change, R32W, while the other three changes are single nucleotide polymorphisms (SNPs). The R32W substitution in CX31 has been previously documented and is speculated to manifest variable penetrance, similar to the polymorphic allele encoding CX26M34T. Over one-third of all samples were also screened with denaturing high-performance liquid chromatography (dHPLC). Seven out of 25 individuals screened were determined to be positive for CX31 sequence variation. Sequence analysis of the 25 individuals screened identified nucleotide alterations in all of the 7 'positives' and in none of the 16 'negatives' yielding a specificity and sensitivity of 100%. Thus, dHPLC represents a highly efficient CX31 screening technique. This study suggests that while sequence alterations are common, pathogenic mutations of CX31 are infrequent in sporadic non-syndromic hearing impairment. PMID- 12630967 TI - Absence of overt iron overload in two individuals compound heterozygotes for a 22 base pair deletion of exon 2 and the C282Y missense mutation of the HFE gene. PMID- 12630970 TI - Complementary medicines, clinical pharmacology and therapeutics. PMID- 12630968 TI - A novel mutation in exon 7 in a family with mild tricho-rhino-phalangeal syndrome type I. PMID- 12630971 TI - Quality, efficacy and safety of complementary medicines: fashions, facts and the future. Part I. Regulation and quality. AB - This is the first of two papers which review issues concerning complementary medicines. The first reviews the extent of use of complementary medicines, and issues related to the regulation and pharmaceutical quality of these products; the second considers evidence for the efficacy of several well-known complementary medicines, and discusses complementary-medicines pharmacovigilance. The term complementary medicines describes a range of pharmaceutical-type preparations, including herbal medicines, homoeopathic remedies, essential oils and dietary supplements, which mainly sit outside conventional medicine. The use of complementary medicines is a popular healthcare approach in the UK, and there are signs that the use of such products is continuing to increase. Patients and the public use complementary medicines for health maintenance, for the treatment or prevention of minor ailments, and also for serious, chronic illnesses. The pervasive use of complementary medicines raises several concerns. Many of these arise because most complementary medicines are not licensed as medicines, and therefore evidence of quality, efficacy and safety is not required before marketing. The regulatory framework for herbal medicines and dietary supplements is currently under review. A new system for registration of traditional herbal medicines will ensure that marketed products meet standards for quality and safety. At present, the pharmaceutical quality of many complementary medicines is a cause for concern. PMID- 12630973 TI - Pharmacokinetics of vanadium in humans after intravenous administration of a vanadium containing albumin solution. AB - AIMS: Vanadium is currently undergoing clinical trials as an oral drug in patients with noninsulin-dependent diabetes mellitus. Furthermore, vanadium occurs in elevated concentrations in the blood of patients receiving intravenous albumin solutions containing large amounts of the metal ion as an impurity. The present study was performed to examine the pharmacokinetics of vanadium in humans following a single intravenous (i.v.) dose of a commercial albumin solution containing a high amount of vanadium. METHODS: The study was conducted in five healthy volunteer subjects who received intravenously 90 ml of a commercial 20% albumin infusion solution containing 47.6 micro g vanadium as an impurity. Vanadium concentrations in serum and urine were determined by electrothermal atomic absorption spectrometry. RESULTS: Vanadium serum concentrations after i.v. administration were measured for 31 days. The data could be fitted by a triexponential function corresponding formally to a three-compartment model. There was an initial rapid decrease in serum concentrations with half-lives of 1.2 and 26 h. This was followed by a long-terminal half-life time of 10 days. The terminal phase accounted for about 80% of the total area under the serum concentration-time curve (AUC). The mean apparent volume of distribution of the central compartment was found to be 10 l. The volume of distribution at steady state was 54 l, and total clearance was 0.15 l h(-1). Vanadium was mainly excreted by the kidneys. About 52% of the dose was recovered in the urine after 12 days. CONCLUSIONS: This study provides data on vanadium pharmacokinetics in healthy humans. PMID- 12630972 TI - Glutathione, glutathione-dependent enzymes and antioxidant status in erythrocytes from children treated with high-dose paracetamol. AB - AIM: To investigate glutathione and antioxidant status changes in erythrocytes from febrile children receiving repeated supratherapeutic paracetamol doses. METHODS: Fifty-one children aged 2 months to 10 years participated in the study. Three groups were studied: group 1 (n = 24) included afebrile children who did not receive paracetamol; and groups 2 (n = 13) and 3 (n = 14) included children who had fever above 38.5 degrees C for more than 72 h. Patients in group 2 received paracetamol at a dose of 50 +/- 15 (30-75) mg kg(-1) day(-1) and those in group 3 received paracetamol above the recommended therapeutic dose, ie 107 28 (80-180) mg kg(-1) day(-1). A blood sample was taken for the measurement of liver transaminases, gammaglutamil transferase (GGT), reduced glutathione (GSH), glutathione reductase (GR), glutathione peroxidase (GPX), glutathione S transferase (GST), superoxide dismutase (SOD) and antioxidant status. RESULTS: Aspartate aminotransferase activity in group 3 was higher than in the other groups (P = 0.027). GSH, SOD and antioxidant status were significantly lower in group 3 compared with groups 1 and 2 (mean differences: for GSH 3.41 micromol gHb(-1), 95% confidence interval (CI) 2.10-4.72, and 2.15 micromol gHb(-1), 95% CI 0.65-3.65, respectively; for SOD 856 U min(-1) gHb(-1), 95% CI 397-1316, and 556 U min(-1) gHb(-1), 95% CI 30-1082, respectively; and for antioxidant status 0.83 mmol l(-1) plasma, 95% CI 0.30-1.36, and 0.63 mmol l(-1) plasma, 95% CI 0.02 1.24, respectively). GR activity was significantly lower in groups 3 and 2 in comparison with group 1 (mean differences 3.44 U min(-1) gHb(-1), 95% CI 0.63 6.25, and 5.64 U min(-1) gHb(-1), 95% CI 2.90-8.38, respectively). Using multiple regression analysis, paracetamol dose was found to be the only independent variable affecting GR, GST and SOD activities (P = 0.007, 0.003 and 0.008, respectively). CONCLUSIONS: In febrile children, treatment with repeated supratherapeutic doses of paracetamol is associated with reduced antioxidant status and erythrocyte glutathione concentrations. These significant changes may indicate an increased risk for hepatotoxicity and liver damage. PMID- 12630974 TI - Comparative effect of human soluble insulin and insulin aspart upon hypoglycaemia induced alterations in cardiac repolarization. AB - AIMS: Sudden death in young diabetic patients has been associated with nocturnal hypoglycaemia perhaps as a result of cardiac dysrhythmias following abnormal cardiac repolarization during hypoglycaemia. It was therefore important to compare the effect of soluble human insulin (HI) and a rapid-acting insulin analogue, insulin aspart (IAsp), on these aspects of cardiac function. METHODS: A total of 17 healthy males underwent identical hyperinsulinaemic hypoglycaemic clamps with blood glucose maintained at 5 mm for 30 min and reduced to 2.5 mm after an additional 30 min. Subjects received either HI or IAsp on two different occasions separated by 4-6 weeks. Regular measurements were made of two measures of cardiac repolarization, QT dispersion and QTc as well as of counter-regulatory hormones. RESULTS: The blood glucose lowering effect did not differ between IAsp and HI and the clearance rates were similar (HI mean +/- SD 1.24 +/- 0.12 l h(-1) kg(-1), IAsp mean +/- s.d. 1.22 +/- 0.32 l h(-1) kg(-1)). There were similar significant increases but no difference between treatments in QTc after hypoglycaemia induced by either IAsp or HI (480 +/- 37 ms vs 480 +/- 25 ms; NS). However, QT dispersion during hypoglycaemia was less pronounced with IAsp than with HI (92 +/- 36 ms vs 107 +/- 42 ms; P < 0.05). Plasma adrenaline increased significantly and similarly after both insulins (initial and final concentration, HI, 0.23 +/- 0.01 to 4.87 +/- 0.48 nm, P < 0.001, IAsp, 0.24 +/- 0.01 to 4.99 +/- 0.48 nm, P < 0.001). Serum potassium decreased significantly but by a similar amount between the groups (initial and final concentration, HI, 4.18 +/- 0.3 to 4.2 +/- 0.2 mm, P < 0.001, IAsp, 4.2 +/- 0.3 to 4.2 +/- 0.3 mm, P < 0.001). CONCLUSIONS: Soluble human insulin and insulin aspart had similar effects upon hypoglycaemia-induced alterations in cardiac repolarization, presumably because the effects of both regular insulin and insulin aspart on the sympathoadrenal response and potassium concentration were the same. PMID- 12630976 TI - The distribution and gender difference of CYP3A activity in Chinese subjects. AB - AIMS: To investigate the distribution of CYP3A activity in the Chinese population, and to test for gender-related differences in CYP3A activity. METHODS: Using midazolam as a probe drug, CYP3A activity in 202 Chinese healthy subjects (104 men) was measured by plasma 1'-hydroxymidazolam:midazolam (1'-OH MDZ:MDZ) ratio at 1 h after oral administration of 7.5 mg midazolam. The different phases of the menstrual cycle including preovulatory, ovulatory and luteal phases of 66 women phenotyped with midazolam were recorded. The concentrations of 1'-OH-MDZ and MDZ in plasma were measured by HPLC RESULTS: A 13 fold variation of CYP3A activity (log1'-OH-MDZ:MDZ: range -0.949-0.203) was shown. The CYP3A activity was normally distributed as indicated by the frequency distribution histogram, the probit plot and the Kolmogorov-Smirnov test (P > 0.05). The CYP3A activity of women was higher than that of men (median: -0.36 vs 0.43, P < 0.05; 95% CI for difference: -0.127, -0.012). There was a significant difference in CYP3A activity between the three phases of the menstrual cycle. The activity was highest in the preovulatory phase and decreased sequentially in the ovulatory and luteal phases (P < 0.05). CONCLUSIONS: A normal distribution of CYP3A activity was observed in the Chinese population. The CYP3A activity is higher in female subjects than in males. CYP3A activity differed across the phases of the menstrual cycle. PMID- 12630975 TI - Population pharmacokinetics and concentration-effect relationships of capecitabine metabolites in colorectal cancer patients. AB - AIMS: To assess the relationship between systemic exposure to capecitabine metabolites and parameters of efficacy and safety in patients with advanced or metastatic colorectal cancer from two phase III studies. METHODS: Concentration effect analyses were based on data from 481 patients (248 males, 193 females; age range 27-86 years) in two phase III studies. Plasma concentration-time data for 5'-deoxy-5-fluorouridine (5'-DFUR), 5-fluorouracil (5-FU) and alpha-fluoro-beta alanine (FBAL) were obtained from sparse blood samples collected within the time windows 0.5-1.5 h, 1.5-3.0 h, and 3.0-5.0 h after capecitabine administration (1250 mg m(-2)) on the first day of cycles 2 (day 22) and 4 (day 64), respectively. Systemic exposure based on plasma concentrations of capecitabine and its metabolites was determined using individual parameter estimates derived from a population pharmacokinetic model constructed for this purpose in NONMEM. Logistic regression analysis was conducted for selected safety parameters (all treatment-related grade 3-4 adverse events, treatment-related grade 3-4 diarrhoea, grade 3 hand-foot syndrome (HFS) and grade 3-4 hyperbilirubinaemia) and for tumour response. Cox regression analysis was used for the analysis of time-to-event data (time to disease progression and duration of survival). RESULTS: Statistically significant relationships between covariates and PK parameters were found as follows. A doubling of alkaline phosphatase activity was associated with a 11% decrease in 5-FU clearance and a 12% increase in its AUC. A 50% decrease in creatinine clearance was associated with a 35% decrease in FBAL clearance, a 53% increase in its AUC, a 24% decrease in its volume of distribution, and a 41% increase in its Cmax. A 30% increase in body surface was associated with a 24% increase in the volume of distribution of FBAL and a 19% decrease in its Cmax. There was a broad overlap in systemic drug exposure between patients regardless of the occurrence of treatment-related grade 3-4 adverse events or response to treatment, leading to weak relationships between systemic exposure to capecitabine metabolites and the safety and efficacy parameters. Of 42 concentration-effect relationships investigated, only five achieved statistical significance. Thus, we obtained a positive association between the AUC of FBAL and grade 3-4 diarrhoea (P = 0.035), a positive association between the AUC of 5-FU and grade 3-4 hyperbilirubinaemia (P = 0.025), a negative association between the Cmax of FBAL and grade 3-4 hyperbilirubinaemia (P = 0.014), a negative association between the AUC of 5-FU (in plasma) and time to disease progression (hazard ratio (HR) = 1.626, P = 0.0056), and a positive association between the Cmax of 5'-DFUR and survival (HR = 0.938, P = 0.0048). Additionally, there were inconsistencies when concentration-effect relationships were compared across the two studies. CONCLUSIONS: Systemic exposure to capecitabine and its metabolites in plasma is poorly predictive of safety and efficacy. The present results have no clinical implications for the use of capecitabine and argue against the value of therapeutic drug monitoring for dosage adjustment. PMID- 12630978 TI - Arylamine N-acetyltransferase-2 genotypes in the Thai population. AB - AIMS: To determine the frequencies of the major arylamine- N-acetyltransferase-2 (NAT2) alleles in the Thai population. METHODS: DNA samples from 235 Thai individuals were analysed by polymerase chain reaction with restriction fragment length polymorphism assays. RESULTS: The frequency distribution of major NAT2 alleles, including NAT2*4, NAT2*5, NAT2*6 and NAT2*7 were 0.381 (95% CI 0.337, 0.426), 0.038 (0.023, 0.060), 0.326 (0.283, 0.370) and 0.204 (0.169, 0.244), respectively. When converted to phenotypes, the study population comprised 63.8% rapid acetylators and 36.2% slow acetylators. CONCLUSIONS: The pattern of NAT2 alleles of Thais is similar to those of many Asian populations, although the frequency of NAT2*4 is significantly lower and NAT2*7 is higher than that of Oriental populations. PMID- 12630977 TI - Phenotyping CYP3A using midazolam in cancer and noncancer Asian patients. AB - AIMS: To investigate CYP3A activity in cancer and noncancer Asian patients using midazolam and to reveal possible alternative traits for phenotyping CYP3A. METHODS: Intravenous midazolam 2.5 mg or 2.5-8 mg was administered to 27 cancer and 24 noncancer patients, respectively. Plasma was sampled at 0, 0.25, 0.5, 1, 1.5, 2, 3.5 and 5 h after intravenous ultrashort, 30 s infusion. Plasma midazolam and 1'-hydroxymidazolam concentrations were determined using GCMS. The disposition of midazolam and 1'-hydroxymidazolam in these patients was compared. Midazolam clearance was correlated with dose-normalized plasma midazolam concentrations (concentration/per dose). RESULTS: Clearance (CL) and steady state volume of distribution (Vss) of midazolam (mean +/- SD, 95% confidence level) in cancer (424 +/- 155, 61.3 ml min(-1); 1.21 +/- 0.46, 0.18 l kg(-1)) and noncancer (407 +/- 135, 57.1 ml min(-1); 1.15 +/- 0.33, 0.155 l kg(-1)) patients, respectively, were not different and comparable with published data. Clearance variability was 4-5 fold in both groups. Midazolam clearance correlated significantly with all plasma concentration/per dose at and after the 1-h time point, with a minimum correlation coefficient of r = 0.752, P < 0.001. CONCLUSIONS: CYP3A activities determined with different doses of midazolam in cancer and noncancer Asian patients showed variability of 4-5-fold and were not different between groups. One to two-fold plasma midazolam concentrations per dose may be feasible as a simple alternative phenotypic trait for hepatic CYP3A activity determination. PMID- 12630979 TI - Aspirin for the prevention of cardiovascular disease: calculating benefit and harm in the individual patient. AB - AIMS: To estimate the absolute reduction in the risk of cardiovascular events and absolute increase in gastrointestinal haemorrhage associated with aspirin for individuals with different baseline risks. METHODS: Calculation of absolute treatment effects from estimates of: (i) baseline risks for cardiovascular event and gastrointestinal haemorrhage; and (ii) relative risks of these events with treatment. Baseline cardiovascular risks were derived from existing risk scores, and baseline risk of gastrointestinal haemorrhage from an observational cohort study. Changes in relative risks were obtained from clinical trial data. The effects of aspirin treatment were calculated in examples of two individuals with very different baseline risks. RESULTS: Treatment of a healthy 74-year-old man (blood pressure 144/88 mm Hg and no history of gastrointestinal disorder) would reduce his annual risk of a cardiovascular event from 2% to 1.74% (absolute risk reduction 0.26%, number needed to treat 385), but increase the gastrointestinal haemorrhage risk from 0.3% to 0.51% (absolute risk increase 0.21%, number needed to harm 476). In a 66-year-old obese man, following a transient ischaemic attack, and with a history of hospital treatment for a peptic ulcer, the annual risk of a cardiovascular event would be reduced from 5% to 4.35% (absolute risk reduction 0.65%, number needed to treat 153), but the risk of gastrointestinal haemorrhage would increase from 1.08% to 1.83% (absolute risk increase 0.75%, number needed to harm 133). CONCLUSIONS: Estimating benefit and harm by taking into account the baseline risks in each individual allows patients and doctors to judge for themselves the magnitude of the trade-offs involved in taking aspirin. PMID- 12630981 TI - Planning and monitoring of placebo-controlled survival trials: comparison of the triangular test with usual interim analyses methods. AB - AIMS: For ethical and economic reasons, interim analysis of phase III clinical trials is essential. This study was conducted to compare the efficiency of two interim analysis procedures, which could be used to allow early termination of a clinical trial. METHODS: We made a post hoc application of two interim analysis methods (Lan & DeMets with O'Brien-Flemming modification, and the triangular test according to Whitehead) by using individual patient data from four published placebo-controlled survival trials. We determined the date the trial would have been stopped had each method been used, and we estimated consequent results in terms of events and patient numbers included in the trial, the duration of the trial, and on treatment effect. RESULTS: The triangular test provided the lowest number of events required to reach a conclusion of the trials while providing an accurate estimate of experimental treatment effects. The triangular test thus indicated the smallest number of patients that would have been enrolled, and the shortest trial duration. The difference between the methods was most important with a detrimental effect of experimental treatment: the number of required events was reduced by 75% and the trial duration was shortened by 48% with the triangular test compared to the Lan & DeMets method. CONCLUSIONS: Stopping a trial early must depend on the clinical context. It is most important to stop a placebo-controlled trial as soon as possible when the experimental treatment can be shown deleterious. In such a situation the triangular test is more appropriate than the Lan & DeMets method. When a treatment effect is no different from, or better than, placebo the triangular test is also superior but the importance of premature termination of the trial in such cases has to be balanced against the inevitable reduction of information that the trial can provide. PMID- 12630980 TI - Who receives lipid-lowering drugs: the effects of comorbidities and patient characteristics on treatment initiation. AB - AIMS: Little is known about the effects of comorbidities and patient characteristics on treatment initiation of lipid-lowering drugs (LLDs), which can be helpful in the evaluation of the risks and benefits of LLDs. METHODS: Baseline characteristics among subjects who received their first ever-recorded LLD prescription in general practice between 1 January 1990 and 31 December 1997, and hyperlipidaemic patients without LLD therapy during the same period were obtained from the UK General Practice Research Database. Differences between patients who received and patients who did not receive LLDs, as well as patients who received different classes of LLDs were compared by fitting multivariate logistic regression models that adjusted for age, sex, body mass index, smoking status, and year of treatment initiation or hyperlipidaemia diagnosis. RESULTS: We found that there were many differences in the baseline characteristics, such as number of general practitioner visits, diagnosis and severity of cardiovascular diseases, and concurrent medications, between the 25 331 patients who received and the 16 287 patients who did not receive LLDs. We also noted that patients with statin therapy had more prior hospitalization, more recent myocardial infarction/stroke, and more concurrent cardiovascular medications, than those patients who received other LLDs. CONCLUSIONS: Patients who received LLDs in primary care, especially patients with statin therapy, were more likely to be elderly and to have more concomitant severe cardiovascular comorbidities than those hyperlipidaemic patients who did not receive LLDs. Examining the medical records of individuals eligible for LLD therapy is an important first step in selectively targeting who will experience the greatest benefit to risk ratio for the treatment of hyperlipidaemia, and is an important step in avoiding confounding by indication when designing epidemiological studies comparing the risks and benefits of treatments for hyperlipidaemia. PMID- 12630982 TI - Low blood selenium concentrations in schizophrenic patients on clozapine. AB - AIMS: To compare plasma and red-cell selenium concentrations of schizophrenic patients treated with clozapine, with healthy controls and patients with mood disorders. METHODS: Plasma and red-cell selenium concentrations were measured in random venous blood samples from four groups: mood disorder (n = 36), schizophrenics treated with clozapine (n = 54), schizophrenics not treated with clozapine (n = 41) and a healthy control group (n = 56). Assays were performed by an independent laboratory that was blinded to the patient groups and specializes in estimating trace metal concentrations. RESULTS: Selenium concentrations in plasma and red cells were found to be significantly lower in schizophrenic patients treated with clozapine as compared with all other groups. CONCLUSIONS: Selenium is an essential antioxidant. Its deficiency has been implicated in myocarditis and cardiomyopathy. Low selenium concentrations in clozapine-treated patients may be important in the pathogenesis of life threatening cardiac side effects associated with clozapine. Further clinical studies are being conducted to explore this important clinical observation and its therapeutic implications. PMID- 12630983 TI - Systemic bioactivity of intranasal triamcinolone and mometasone in perennial allergic rhinitis. AB - AIMS: To evaluate the systemic bioactivity of triamcinolone acetonide (TA) 220 micro g or mometasone furoate (MF) 200 micro g over 3 weeks in perennial allergic rhinitis. METHODS: Twenty-seven patients received TA 220 micro g or MF 200 micro g once daily for 3 weeks with a 2 week placebo washout period prior to each randomized treatment. Measurements were made at baseline after each washout and after each randomized treatment, comprising overnight 10-h urinary cortisol corrected for creatinine (OUCC), 08.00 h plasma cortisol and 08.00 h serum osteocalcin. RESULTS: There were no significant differences between baseline values prior to TA or MF, and for any outcome measures comparing randomized treatments to respective baseline values or comparing TA with MF. For OUCC compared with baseline, the geometric mean fold suppression (95% CI) was 1.02 (0.78, 1.33) for TA (2% decrease), 1.07 (0.80, 1.42) for MF (7% decrease), and 1.05 (0.79, 1.39) for TA vs MF (5% decrease). CONCLUSIONS: Standard doses of TA or MF over 3 weeks showed no differences in systemic bioactivity markers compared with respective baseline values after placebo washout, and there were no differences between TA vs MF. PMID- 12630984 TI - A placebo-controlled pharmacodynamic and pharmacokinetic interaction study between tamsulosin and acenocoumarol. AB - AIM: To evaluate pharmacokinetic and pharmacodynamic interactions between tamsulosin and acenocoumarol. METHODS: Twelve healthy volunteers received tamsulosin 0.4 mg or placebo once daily for 9 days in a double-blind, cross-over study. On day 5 of each study period, a single 10-mg oral dose of racemic acenocoumarol was administered. RESULTS: The ratios (point estimates (90% confidence intervals)) of values in the presence and absence of tamsulosin were: AUCPT 1.01 (0.98, 1.03); maximum prothrombin time (Ptmax) 0.99 (0.94, 1.05); AUC (R)-acenocoumarol 1.02 (0.90, 1.16), and AUC (S)-acenocoumarol 1.03 (0.89, 1.20). Both combinations, tamsulosin and placebo with acenocoumarol, were well tolerated. CONCLUSIONS: Multiple doses of tamsulosin had no effect on the pharmacokinetics or pharmacodynamics of a single high dose of acenocoumarol. PMID- 12630985 TI - Development, implementation and cost-effectiveness of a protocol for review of combination diuretic prescribing. AB - AIMS: To determine the extent of inappropriate prescribing of combination diuretics and the cost implications of protocol implementation. METHODS: Pharmacist-run medication review clinics in two general practices in Bradford, UK. RESULTS: Sixty-one patients, mean +/- s.d. age 75.8 +/- 12.0 years, were reviewed. Thirty-six met protocol criteria; 30 (49.2%) patients had their potassium-sparing diuretics (PSDs) discontinued. Mean (95% confidence interval) reduction in serum potassium concentrations after intervention in these 30 patients was 0.26 (0.09, 0.43) mmol l(-1) (P < 0.01). Twenty-eight (93.3%) patients were within the reference range at follow-up; none was below. CONCLUSIONS: Of prescribed PSDs, 59% were found to be unnecessary. Using the developed protocol potentially improves drug safety and enables cost savings. PMID- 12630986 TI - Genotyping of four genetic polymorphisms in the CYP1A2 gene in the Egyptian population. AB - AIMS: The goal of this study was to determine the frequencies of CYP1A2*1C, *1D, *1E and *1F variants in the Egyptian population and compare frequencies with other populations. METHODS: Genotyping was performed in a total of 212 unrelated Egyptian subjects using polymerase chain reaction-restriction fragment length polymorphism assay. RESULTS: The frequencies of CYP1A2*1C, *1D, *1E and *1F variants in the Egyptian population were 0.07, 0.40, 0.03 and 0.68, respectively. The Egyptians have a lower frequency of CYP1A2*1C, and CYP1A2*1E than the Japanese (0.07 vs 0.21 and 0.03 vs 0.08, respectively), while the frequencies of CYP1A2*1D and CYP1A2*1F did not differ significantly between the two groups. CYP1A2*1F (0.68) frequency in Egyptians was identical to that observed in Caucasians (0.68 among 236 German individuals). CONCLUSIONS: The present study is the first to describe the frequencies of four known allelic variants of CYP1A2 among the Egyptian population. CYP1A2*1C and *1E occurred at frequencies significantly lower than that in Japanese, while similar frequencies were observed for CYP1A2*1D and *1F. The CYP1A2*1F frequency appeared to be identical to that of Caucasians. This does not exclude the possibility of the presence of new mutations relatively specific to the Egyptian population that have not been identified. PMID- 12630987 TI - Concomitant overdosing of other drugs in patients with paracetamol poisoning. PMID- 12630992 TI - Transcatheter embolization of biopsy-related vascular injuries in renal allografts. Long-term technical, clinical and biochemical results. AB - PURPOSE: Biopsy-related vascular injuries in renal transplants are rare, but they can lead to dramatic clinical symptoms prompting immediate treatment. Transcatheter embolization is a known minimally invasive technique to treat some form of arterial bleeding. This study evaluated the efficacy of this technique in iatrogenic biopsy-related vascular lesions in renal allografts. MATERIAL AND METHODS: Between 1993 and 2001, 13 patients were treated by percutaneous transcatheter intervention. Indications for embolization were hypovolemic shock due to perinephric hematoma (n = 5), persistent macroscopic hematuria (n = 7) and an asymptomatic large intrarenal pseudoaneurysm in 1 patient. Selective angiography revealed an arteriovenous fistula (n = 7), a pseudoaneurysm (n = 6), and perinephric contrast extravasation (n = 4). RESULTS: In all patients, successful embolization of the feeding artery could be performed; in 11 patients it was performed in one session, in 2 patients an additional session was needed. In 1 patient thrombosis of a segmental artery occurred immediately after embolization, but was successfully treated by short-term in situ thrombolysis. Clinical symptoms disappeared in all patients. Serum creatinine levels (determined 30 and 60 days after embolization, compared to the level before embolization) decreased significantly in 10 patients; a progressive deterioration of the renal function was observed in 3 patients. CONCLUSIONS: Transcatheter embolization is a safe and effective endovascular technique to treat biopsy related vascular injuries in renal transplants. In the vast majority of cases an immediate clinical success and significant benefit in renal function can be obtained and the longevity of the allograft after successful embolization mainly depends on the natural (medical) outcome. PMID- 12630991 TI - Doppler examination of the allografted kidney. AB - A comprehensive ultrasound examination of the transplanted kidney includes a Doppler examination. Duplex Doppler, color Doppler and power Doppler can all reveal important information. In addition, calculation of the resistance and pulsatility indices to quantify changes in the spectral Doppler waveform can be of great help, particularly in the first weeks and months following a transplantation. The Doppler part of the examination should evaluate the vessels to and from the transplant, as well as the parenchyma with calculations of indices to detect the presence of increased vascular resistance. The conclusions drawn from the Doppler-derived information combined with the results from gray scale scanning and clinical information will very often be of clinical significance for the handling of the patient. PMID- 12630993 TI - Echo-enhanced color Doppler cystosonography of vesicoureteral reflux in children. Improvement by stimulated acoustic emission. AB - PURPOSE: To prospectively evaluate the potential of echo-enhanced color Doppler sonography (ee-CDS) using the acoustic stimulated emission technique in the diagnosis of vesicoureteral reflux (VUR) in children. MATERIAL AND METHODS: 30 children (age: 0-9 years) with suspicion of VUR were submitted for echo-enhanced cystosonography (ee-CS) and conventional voiding cysto-urethrography (VCU). Conventional US and then color Doppler sonography (CDS) were performed after instillation of Levovist into the catheterized urinary bladder that had been prefilled with saline. Finally, output gain was increased in order to 'burst' the bubbles (= stimulated acoustic emission, burst-CS). Thereafter VCU was performed in the same session. The results of conventional ee-CS and burst-CS were compared to VCU findings. RESULTS: Twenty-four refluxing systems were found in a total of 66 collecting systems by both techniques. Ee-CS and burst-CS detected VUR in four systems not seen by VCU (grades 1-4), but missed low-degree VUR in 2 cases. In low-degree VUR, burst-CS was particularly helpful in 9 patients with equivocal results on conventional ee-CS. In 6 of them, VUR was proven by burst-CS, the other 3 showed VURs neither on VCU nor on burst-CS. CONCLUSION: Levovist ee-CS is feasible in the pediatric bladder. It enables reliable visualization of VUR in the renal collecting system. Burst-CS improves visualization of contrast reflux, particularly in non-dilative or minimally dilative VUR, which is sometimes difficult to depict with conventional ee-CS. PMID- 12630994 TI - Testicular tumors manifested as inferior vena cava thromboses. Case reports. AB - Four cases of asymptomatic testicular tumors with inferior vena cava (IVC) involvement are reported. All patients were referred to our hospital with non specific abdominal pain and abdominal US revealed an inferior vena cava thrombosis. CT showed an IVC thrombus extending from the first to second lumbar vertebrae and also retroperitoneal lymphadenopathy in 3 patients. Scrotal US demonstrated intratesticular tumors. IVC thrombosis may result from asymptomatic intratesticular tumors (single or multiple). Because of that, scrotal US is of paramount importance as a routine screening test in patients who radiographically demonstrate caval thrombosis. PMID- 12630995 TI - Comparison of 99mTc-sestamibi scintimammography and dynamic MR imaging as adjuncts to mammography in the diagnosis of breast cancer. AB - PURPOSE: To compare the diagnostic accuracy of planar 99mTc-sestamibi scintimammography with dynamic contrast-enhanced MRI (CE-MRI) on the basis of histopathologic results, and to determine the clinical value of these methods as adjuncts to mammography. MATERIAL AND METHODS: A total of 90 consecutive women with 111 histopathologically verified breast lesions were enrolled in the study. Patients underwent scintimammography and CE-MRI in addition to mammography. Each finding was classified on a BI-RADS-like five-point rating scale describing the degree of suspicion for malignancy, and all findings were correlated with the histopathological results. RESULTS: The overall sensitivity/specificity/accuracy was 85%/59%/78% for mammography, 94%/47%/80% for CE-MRI, and 82%/75%/80% for scintimammography, respectively. CE-MRI showed higher sensitivity (p = 0.008), but its specificity was lower than scintimammography (p = 0.049). Using ROC analysis, significant improvement ( p = 0.034) was found between mammography and the combination of mammography + CE-MRI, while mammography + scintigraphy showed no higher diagnostic accuracy than mammography alone. CONCLUSION: If high sensitivity and spatial resolution are needed, CE-MRI is to be preferred in clinical practice as an adjunct to mammography, rather than scintigraphy. PMID- 12630996 TI - Localization of breast sentinel lymph nodes by MR lymphography with a conventional gadolinium contrast agent. Preliminary observations in dogs and humans. AB - PURPOSE: To test the capability of an indirect MR lymphography (MRLG) using a conventional extracellular gadolinium (Gd)-based contrast agent for localizing breast sentinel lymph node (SLN). METHODS: A total of 1 and 0.5 ml of undiluted gadopentetate dimeglumine were injected subcutaneously into the two periareolar areas overlying each caudal mammary gland in 10 female dogs. Contiguous 2-mm thick fast gradient echo MR images were acquired through the upper breast and axilla before and for 60 min after gentle massage at the injection sites, yielding 3D displays. The localized SLN was resected from the living animals, followed by post mortem examinations. MRLG (1 ml contrast agent injected) was also attempted in 5 female volunteers. RESULTS: Even with 0.5-ml contrast agent, the MRLG clearly visualized the connection of SLN and lymphatic vessels directly draining from the injection sites, and the remaining distant nodes, with the maximal enhancement peaks within 10 min after massage. The 3D images provided comprehensive anatomy of these lymphatic pathways in each axilla. Of the 20 SLN and 128 distant nodes visualized on the MRLG, all the SLN (100%) and 107 (83.5%) distant nodes could be resected pre- and post mortem, in good correlation with the locations and sizes on the MR images. MRLG also effectively localized SLN in the volunteers, without significant adverse effects. CONCLUSION: An indirect MRLG using small volumes of conventional Gd-based contrast agent may have potential for accurate identification and surgical biopsy of breast SLN. PMID- 12630997 TI - Computer-aided preoperative diagnosis of microcalcifications on mammograms. AB - PURPOSE: To evaluate of a computer-aided method for differentiating malignant from benign clustered microcalcifications. MATERIAL AND METHODS: Our material was 350 suspicious microcalcifications on mammograms from 330 female patients who underwent breast biopsy (after hook wire localization and under mammographic guidance). The histologic findings were malignant in 140 cases (40%) and benign in 210 cases (60%). Those clusters were manually detected, computer-aided analyzed and quantitatively estimated. Besides computer analysis, 3 physicians observers (2 radiologists and 1 breast surgeon) evaluated the malignant or benign nature of the clustered microcalcifications. The performance of the artificial network, each observer and the three observers as a group was evaluated by receiver operating characteristics (ROC) curves. RESULTS: Comparison of the ROC curves revealed the following AUC values (area under the curve): computer - 0.950, physician 1 - 0.815, physician 2 - 0.830, physician 3 - 0.830, and physicians as a group - 0.825. The results, compared by the student t-test for paired data, showed a statistically significant difference between computer analysis and physicians' performance, independently and as a group. CONCLUSION: Our study showed that computer analysis achieved statistically significantly better performance than that of physicians in the classification of malignant and benign calcifications. PMID- 12630998 TI - Semiautomatic core biopsy. A modified biopsy technique in breast diseases. AB - PURPOSE: To evaluate a semiautomatic gun to retrieve diagnostic core specimens of lesions in the breast and the axillary region. MATERIAL AND METHODS: In a series of 180 consecutive core biopsies (2.1 mm), 145 (142 breasts and 3 axillae) were performed with a semiautomatic gun (18 stereotaxic and 127 US-guided) from lesions presenting mammographically as microcalcifications (n = 15) and opacities (n = 130). The gun did not work satisfactorily in 34 lesions, which were tumors with a very hard consistency. One additional patient was excluded because of technical failure. Biopsy diagnoses in the 145 patients were correlated to surgical histopathology, follow-up mammograms and/or clinical findings. RESULTS: Histologic examination of the specimens resulted in correct diagnoses in 89% (129/145) of the total material and in 87% (108/124) of cancers. In 107 cases, in which only 1 specimen was obtained, 83 of 89 cancers (87%) were detected. Length of specimens ranged from 3 mm (n = 2) to 17 mm (n = 31). Among patients with a 17 mm-long specimen, there was only 1 false-negative diagnosis. CONCLUSION: The semiautomatic gun provided diagnostic specimens in a majority of cases and could be used as an alternative to the automatic guns when size or location of the lesions necessitates a high precision. It was not suitable for use in very hard tumors. PMID- 12630999 TI - Patient positioning after fine needle lung biopsy-effect on pneumothorax rate. AB - PURPOSE: To evaluate the efficacy of precautionary positioning in preventing the development of pneumothorax and reducing the rate of chest tube placement after fluoroscopy-guided transthoracic needle biopsy (TTNB). MATERIAL AND METHODS: One hundred and seven patients who underwent TTNB were randomly assigned to one of two postbiopsy treatment groups: patients were placed with (precautionary positioning group) and without (non-precautionary positioning group) recumbent puncture site either down (n = 59 and 48, respectively) for at least 30 min. RESULTS: The overall pneumothorax rate was 8.4% (n = 9). Four of the 9 patients needed chest tube insertion. Pneumothorax rate was 8.33% (4/48) and 8.47% (5/59) in the precaution and non-precaution groups, respectively. Rate of chest tube insertion was 6.25% (n = 3) and 1.85% (n = 1) in the precaution and non precaution groups, respectively. There was no statistically significant difference in the rate of pneumothorax and chest tube insertion between the two groups. CONCLUSION: Precautionary positioning after TTNB does not reduce the incidence of postbiopsy pneumothorax and rate of chest tube insertion. PMID- 12631000 TI - Do radiologists want/need training in cardiopulmonary resuscitation? Results of an Internet questionnaire. AB - PURPOSE: Prompt and effective cardiopulmonary resuscitation (CPR) decreases morbidity and mortality following cardiopulmonary arrest. Radiologists are frequently confronted with severely ill patients, who may deteriorate at any time. Furthermore, they have to be aware of life-threatening reactions towards contrast media. This study was designed to assess experience and self-estimation of German-speaking radiologists in CPR and cardiac defibrillation (CD). MATERIAL AND METHODS: 650 German-speaking radiologists were audited by a specially designed questionnaire, which was sent via e-mail. The answers were expected to be re-mailed within a 2-month period. RESULTS: The response rate was 12.6%. 72.8% of the responders had performed at least 1 CPR (range 9.5 +/- 13.1) and 37% at least 1 CD. 67.9% had had opportunities to attend training courses, which had been utilized by 41.8% of them. The last training of the responders was more than 2 years ago in 69.2% and more than 5 years ago in 37%. Of all responders 75.6% expressed the need for further education. CONCLUSION: The small response rate indicates the small importance of CPR in the subpopulation surveyed. The vast majority of the responders, however, showed interest in basic and advanced life support and advocated regular updates. It seems reasonable that radiological departments themselves should organize courses in order to cope with their specific situations. PMID- 12631001 TI - Peripheral arterial disease. Contrast-enhanced 3D MR angiography of the lower leg and foot compared with conventional angiography. AB - PURPOSE: To determine whether contrast-enhanced 3D MR angiography (CE MRA) could replace digital subtraction angiography (DSA) for the evaluation of atherosclerotic peripheral vascular disease of the lower leg and foot. MATERIAL AND METHODS: Thirty-five patients with symptoms of atherosclerotic disease of the leg were examined prospectively with CE MRA of the foot and the lower legs as well as with DSA from the aorta to the pedal arches. The MRA technique was focused on optimal imaging of the arteries of the foot. RESULTS: The agreement between CE MRA and DSA for grading of stenosis was moderate to good (weighted kappa-values 0.48-0.80). The sensitivity of CE MRA for detection of significant stenosis (> or = 50%) was 92% and the specificity was 64% with DSA as gold standard. CONCLUSION: CE MRA is a fairly accurate method for the demonstration of atherosclerotic disease below the knee including the pedal arches. It can replace DSA for the assessment of distal arteries in patients with impaired renal function. However, image quality and resolution still needs to be improved before CE MRA can become the method of choice in all patients. PMID- 12631002 TI - Osteochondritis dissecans of the hip. AB - PURPOSE: To investigate the clinical and radiological characteristics of osteochondritis dissecans (OD) of the hip and the outcome of this condition after treatment. MATERIAL AND METHODS: Twelve male and 3 female patients with OD were retrospectively studied. Six patients had a history of Legg-Calve-Perthes disease (LCPD) and among them 2 also had had a trauma to the hip. A further 5 had had trauma and 1 a developmental dislocation of the hip (DDH). The remaining 3 patients had no history of previous hip disease or trauma. All patients were examined with plain radiography, 7 with MR, 3 with CT and 2 with hip arthrography. RESULTS: All OD lesions were detected at plain radiography, and most of them were located near the fovea. At MR the lesions had low signal intensity at T1-weighted sequences, and 6/7 had edema or fluid collection in or adjacent to the lesion on T2-weighted sequences. The early treatment in 7 patients was surgery, 2 had had conservative treatment and 6 no treatment. At follow-up 12 years after radiological diagnosis, 5 patients had hip arthrosis, 4 of whom were treated with arthroplasty. All but 3 had reduced hip rotation and all but 2 (with arthroplasty) had load pain. Three of the patients with earlier surgery had not developed arthrosis. CONCLUSIONS: OD lesions are usually well seen with plain radiography. There is a great risk of developing early arthrosis and it seems that early surgery is connected with arthrosis development. Thus only symptomatic treatment is recommended. PMID- 12631003 TI - US assessment of hip joint synovitis in rheumatic diseases. A comparison with MR imaging. AB - PURPOSE: To assess the significance of ultrasonography (US) in detecting hip joint synovitis in patients with rheumatic diseases. MATERIAL AND METHODS: Forty patients with rheumatic disease and suspected hip joint synovitis underwent MRI and US of the hip joint. In addition to the throughout MRI evaluation, the anterior collum-capsule distance (CCD) was determined by both MRI and US. Thirteen healthy volunteers were examined with MRI to establish the criteria for normal findings in MRI when classifying hip joints to those with synovitis and those without. MRI was used as a gold standard. RESULTS: Synovitis was found using MRI in 31 hips of 22 patients (9 patients had bilateral synovitis). The intraclass correlation was 0.61 between MRI and US in measuring CCD. In classifying hip joint synovitis with US, the sensitivity of the method was 87% and specificity 42%, when the CCD criterion for synovitis was determined to be > or = 7 mm. If the cut-off point was raised to 9 mm, the sensitivity decreased to 61% while specificity increased to 94%. A difference in CCD of > or = 1 mm between the hips as an additional criterion for synovitis increased the number of false-positive findings. CONCLUSION: Measurement of CCD with US proved to be a rather inaccurate method to point out synovitis in rheumatic patients when using MRI as a reference. The main reason for this result was the thickened capsule, which US could not differentiate from a thickened synovium. PMID- 12631004 TI - Grading scale of radiographic findings in the pubic bone and symphysis in athletes. AB - PURPOSE: Radiographic abnormalities in the pubic bone and symphysis are often seen in athletes with groin pain. The aim was to create a grading scale of such radiologic changes. MATERIAL AND METHODS: Plain radiography of the pelvic ring including the pubic bone and the symphysis was performed in 20 male athletes, age 19-35, with long-standing uni- or bilateral groin pain. We used two control groups: Control group 1: 20 healthy age-matched men who had undergone radiologic examination of the pelvis due to trauma. Control group 2: 120 adults (66 men and 54 women) in 9 age groups between 15 and 90 years of age. These examinations were also evaluated for interobserver variance. RESULTS AND CONCLUSION: The grading scale was based on the type and the amount of the different changes, which were classified as follows: No bone changes (grade 0), slight bone changes (grade 1), intermediate changes (grade 2), and advanced changes (grade 3). The grading scale is easy to interpret and an otherwise troublesome communication between the radiologist and the physician was avoided. There was a high interobserver agreement with a high kappa value (0.8707). Male athletes with long-standing groin pain had abnormal bone changes in the symphysis significantly more frequently and more severely (p>0.001) than their age-matched references. In asymptomatic individuals such abnormalities increased in frequency with age both in men and women. PMID- 12631005 TI - Spatial component position in total hip arthroplasty. Accuracy and repeatability with a new CT method. AB - PURPOSE: 3D detection of centerpoints of prosthetic cup and head after total hip arthroplasty (THA) using CT. MATERIAL AND METHODS: Two CT examinations, 10 min apart, were obtained from each of 10 patients after THA. Two independent examiners placed landmarks in images of the prosthetic cup and head. All landmarking was repeated after 1 week. Centerpoints were calculated and compared. RESULTS: Within volumes, all measurements of centerpoints of cup and head fell, with a 95% confidence, within one CT-voxel of any other measurement of the same object. Across two volumes, the mean error of distance between center of cup and prosthetic head was 1.4 mm (SD 0.73). Intra- and interobserver 95% accuracy limit was below 2 mm within and below 3 mm across volumes. No difference between intra- and interobserver measurements occurred. A formula for converting finite sets of point landmarks in the radiolucent tread of the cup to a centerpoint was stable. The percent difference of the landmark distances from a calculated spherical surface was within one CT-voxel. This data was normally distributed and not dependent on observer or trial. CONCLUSION: The true 3D position of the centers of cup and prosthetic head can be detected using CT. Spatial relationship between the components can be analyzed visually and numerically. PMID- 12631006 TI - Quantitative diagnosis of fatty liver with dual-energy CT. An experimental study in rabbits. AB - OBJECTIVES: To explore the correlation between fatty content of fatty liver and the difference of CT attenuation value in dual-energy CT, and to evaluate the value of dual-energy CT in the quantitative diagnosis of fatty liver in rabbits. MATERIAL AND METHODS: Dual-energy CT at 120 kVp and 90 kVp was performed in 16 rabbits of experimental groups that were induced to various degrees of fatty liver by feeding carbon tetrachloride (CCl4). Four healthy rabbits in the control group underwent dual-energy CT at the same time. The CT attenuation values of liver tissues at 120 kVp (H120), 90 kVp (H90) and the difference (Delta H) between them were obtained. The specimens of liver tissues were examined with freeze section (9 microm) and an oil red O stain histologically. The ratio of fatty content in liver to the liver volume (VP value) was measured using the image analyzer system. RESULTS: The change of CT attenuation values between 120 kVp and 90 kVp was not obvious (Delta H between -0.4 HU and 1.4 HU) in the control group. The density of the liver in the experimental groups decreased differently, and the change in CT attenuation values between high and low energy levels were very clear. H120 values were between 51.0 HU and 91.7 HU and H90 values were between 37.0 HU and 89.2 HU. Delta H values were between 2.5 HU and 14.0 HU. Significant negative linear correlation (r = -0.92, -0.93, t = 8.51, 9.76, p < 0.001, 0.001, respectively) were found between VP values and H120 and H90, respectively, while significant positive linear correlation (r = 0.95, t = 11.89, p < 0.001) was found between VP values and Delta H. CONCLUSION: The amount and degree of fat in the fatty liver can be assessed exactly by dual-energy CT. It will be a potential new effective method for quantitative diagnosis of fatty livers. PMID- 12631007 TI - Preoperative right portal vein embolization in patients with metastatic liver disease. Metastatic liver volumes after RPVE. AB - PURPOSE: To quantify liver metastases and future remnant liver (FRL) volumes in patients who underwent right portal vein embolization (RPVE) and to evaluate the effects of this procedure on metastase growth. MATERIAL AND METHODS: Nine patients with liver metastases from primary colon (n = 5), rectal lesions (n = 1) and carcinoid tumors (n = 3) underwent spiral CT to evaluate the ratio of the non tumorous parenchymal volume of the resected liver to that of the whole liver volume (R2). Hand tracing was used to isolate the entire liver, the resected liver and metastase volumes. All patients with R2 > 60% underwent RPVE. RESULTS: FRL exhibited a 101-336 cm3 (average 241 cm3) increase in volume 1 month after RPVE. One patient refused surgery for 2 months and before surgery the increase in volume of the FRL was similar to that of other patients (180.64 cm3). Percent metastases volume from colorectal carcinoma in embolized liver parenchyma increased from 62.4% to 138.4% at 1 month and to 562% at 2 months after RPVE. Metastase volume from carcinoid tumors was unchanged. CONCLUSION: One month after RPVE, hypertrophy of the FRL is evident. In the embolized liver, there was a progressive increase in metastase volume from colorectal carcinoma while metastase volume from carcinoid tumor was unchanged in embolized and non embolized liver. PMID- 12631008 TI - Contrast-enhanced transabdominal ultrasonography in the diagnosis of pancreatic mass lesions. AB - PURPOSE: In this study, we evaluated the diagnostic usefulness of contrast enhanced transabdominal ultrasonography (CE-US) in pancreatic mass lesions that could be detected by US. MATERIAL AND METHODS: CE-US was performed in 37 patients with various pancreatic mass lesions to evaluate the characteristic vascular findings. RESULTS: Pancreatic tubular adenocarcinoma showed negative enhancement pattern in 16 cases and a slight spotty enhancement pattern in 5 cases. Poorly differentiated adenocarcinoma showed slight homogeneous enhancement pattern in 1 case and acinar cell carcinoma was expressed as a heterogeneous enhancement pattern in 3 cases on CE-US. Endocrine cell tumor had a strong homogeneous enhancement pattern in 3 cases, a strong central enhancement in 1 case and a strong peripheral enhancement in 2 cases. Pseudotumorous pancreatitis showed an iso-enhancement pattern in 6 cases. CONCLUSION: CE-US is useful in evaluating the vascularity of the lesions in various pancreatic mass lesions in real-time images, and may be useful for the differential diagnosis of pancreatic mass lesions. PMID- 12631009 TI - Multidetector-row CT appearance of acute torsion of wandering spleen in a child. PMID- 12631010 TI - MR imaging of subclinical cerebral decompression sickness. A case report. AB - Diving accidents related to barotrauma constitute a unique subset of ischemic insults to the central nervous system. Victims may demonstrate components of arterial gas embolism, which has a propensity for cerebral involvement, and/or decompression sickness, with primarily spinal cord involvement. Decompression sickness-related radiology literature is very limited. We present our MR findings including FLAIR images in a decompression sickness patient with atypical presentation and review the related literature. We believe MR can be useful in follow-up studies and in early diagnosis of decompression sickness when symptoms do not fit the classic picture or loss of consciousness in surfacing. PMID- 12631011 TI - Et-A receptor antagonist BQ123 prevents radiocontrast media-induced renal medullary hypoxia. AB - PURPOSE: Renal vasoconstriction with resultant tissue hypoxia, especially in the renal medulla, has been suggested to play a role in contrast media (CM)-induced nephropathy. Endothelin (ET) is released into the blood stream following CM injection and has been proposed as a potential mediator through its vasoconstrictive properties. MATERIAL AND METHODS: To investigate the possible protective influence of ET-receptor antagonists against CM-induced reduction in renal function, we studied the effects of injection of iopromide with and without pretreatment with BQ123 (ET-A antagonist) or BQ788 (ET-B antagonist) on renal superficial cortical flow (CBF), outer medullary blood flow (OMBF) and outer medullary oxygen tension (pO2) in normal rats. RESULTS: Administration of CM (1600 mg I/kg b.w.) did not affect CBF in any of the groups. However, a transient decrease in OMBF occurred, which was unaffected by both BQ123 and BQ788. Also a transient decrease in outer medullary pO2 was induced by CM administration. The pO2 reduction was significantly smaller after pretreatment with BQ123, than after injection of CM alone or together with BQ788, and pO2 returned more rapidly to the control level. Neither receptor antagonist had an effect on CM-mediated increases in electrolyte excretion. CONCLUSION: In the normal rat, activation of ET-A receptors is partly involved in the depression of outer medullary pO2 caused by injection of iopromide. However, the decrease in OMBF after iopromide injection is not mediated by ET receptors. The beneficial effects of the ET-A receptor antagonist on CM-induced changes in outer medullary pO2 seem therefore not primarily mediated on the hemodynamic level but may rather involve tubular transport mechanisms. PMID- 12631013 TI - An evidence-based glaucoma guideline. PMID- 12631014 TI - The Finnish evidence-based guideline for open-angle glaucoma. AB - In most patients, chronic open-angle glaucoma is a slowly progressive disease. Eyes with very high intraocular pressure (IOP > 30 mmHg) represent an exception to this and should be treated and followed extremely intensively. As lowering IOP is, so far, the only means of treating glaucoma, the majority of research reports deal with the IOP-lowering effect of the treatment. The primary goal of treatment, however, is to prevent glaucomatous damage to the structures and function of the eye. The effectiveness of treatment is monitored with optic disc and retinal nerve fibre layer imaging and with visual field examinations. If the glaucomatous changes are progressing, more effective treatment should be given. In the course of follow-up, it should be noted that the changes in the optic nerve structure and function appear and progress at different time-points with delays of up to several years. The assessment of abnormalities is dependent on the examination method and requires a great deal of experience on the part of the examiner. The important risk factors in glaucoma are elevated IOP (even if IOP is within normal range in half of patients ), age, positive family history, exfoliation, race and myopia. PMID- 12631015 TI - Phototherapeutic keratectomy: 12 years of experience. AB - BACKGROUND: Phototherapeutic keratectomy (PTK) has been employed as a surgical tool to treat corneal disease for more than 10 years. The laser has made it possible to remove superficial corneal opacities and thereby restore vision. The 193 nm ultraviolet light separates molecules and splits molecules in biological tissue, thereby ablating it. About 0.25 microm of tissue is ablated by each pulse. The development of the excimer laser technique has been fast. It has principally focused on refractive surgery but has also benefited PTK. CORNEAL DYSTROPHIES: The ability to delay or postpone corneal grafting in superficial corneal dystrophies represents a very important achievement. Map-dot-fingerprint dystrophy or basal membrane dystrophy is a common indication for PTK. Other dystrophies such as Meesman's, Reis-Buckler's, Thiel-Benke's, granular, macular, lattice and Schnyder's can be treated, although with differing degrees of success and varying rates of recurrence. Subepithelial scarring in Fuchs' dystrophy has been ablated. Other trials have involved the removal of substantial parts of the stroma in order to reduce the load on the endothelium. Recurrent dystrophic changes can likewise be removed from corneal grafts and thus prevent the need for regrafting. RECURRENT EROSIONS: Laser treatment has made it possible to manage wound-healing problems better after recurrent erosions. Recurrent erosions are the most common indications for PTK: several studies show good and persistent effects with this type of treatment. Persistent epithelial defects of various origins, among them corneal ulcers resulting from allergic disease, can likewise be treated. SCAR TISSUE: Scars after surgery such as pterygeum excision can be removed. Smooth muscle actin containing fibroblasts in old scars should be given special consideration in PTK. Excimer laser surgery can be successfully combined with conventional surgery to remove excessive scar tissue, Salzmann's nodules and very flaky and coarse band keratopathy. Irregular corneal surfaces following ulcers and injuries pose problems that have so far proved difficult to overcome. Thinning is often seen after bacterial corneal ulcers or after herpes simplex keratitis. A rough or uneven surface can be made smoother by using modulators during treatment by casting a new surface under a hard contact lens (PALM technique), a surface that is then projected into the stroma by laser ablation. Modern techniques linking the excimer laser with computerized corneal topography and wavefront analysis promise to further improve the smoothing capacities of lasers and to increase the quality of optical results. COMPLICATIONS: The most feared complication of PTK is the postoperative infection. These are rare. Haze is usually not prominent but scar tissue formation of a more persistent type has been noted after laser surgery in eyes with pre-existing surgical scars. Keratectasia has been described after PTK. Failure due to deep opacities or a surface that is too uneven is a more common frustration. This paper reviews advances in excimer laser treatment of corneal disease. PMID- 12631016 TI - Astigmatic axis and amblyopia in childhood. AB - PURPOSE: This study is part of a larger project whose aim is to evaluate the relationship between refractive errors and amblyopia. In an earlier study, we showed that there is a substantial increase in the prevalence of amblyopia among children with oblique astigmatism. To further evaluate this relationship, we examined children with astigmatisms of 1 dioptre or more and varying directions of the astigmatic axes. METHODS: Two groups of astigmatic children, with oblique and orthogonal astigmatism, respectively, were selected for the study at 1 year of age via a general health screening programme. The most emmetropic axis was identified in each child and used in the study. Visual acuity (VA) was tested when the children were between 4 and 4.5 years of age. The presence of amblyopia, defined as difference in VA between the eyes of 0.1 log unit or more, and any increase in acuity following occlusion therapy were noted. RESULTS: The angle of the astigmatic axis strongly relates to the risk of developing amblyopia. Axes +/ 15 degrees from the main axes did not affect the risk of amblyopia but oblique astigmatism significantly increased the risk of developing amblyopia (p = 0.0024). CONCLUSIONS: The results accord with earlier findings that oblique astigmatism increases the risk of developing amblyopia. PMID- 12631017 TI - The effects of aspirin and warfarin therapy on haemorrhage in vitreoretinal surgery. AB - BACKGROUND: No substantial recommendations exist regarding the management of anticoagulant drugs prior to ocular surgery. Stopping anticoagulation can cause fatal emboli, but sight-threatening bleeds may occur if anticoagulation is continued. We examined the effects of anticoagulation on vitreoretinal surgery. METHODS: Clinical details were prospectively entered on a database. The anticoagulant status of 541 consecutive patients undergoing vitreoretinal surgery was recorded. RESULTS: Sixty patients in the study were taking aspirin and seven were taking warfarin. There were 11 cases of choroidal haemorrhage, one of which involved a warfarin user. Of 325 retinal detachment repairs, 21 (6.5%) had preoperative vitreous haemorrhages. Two of these patients were on aspirin and two were on warfarin. Sixty-six vitrectomies were performed for diabetic vitreous haemorrhages, of which nine re-bled postoperatively. One of these patients was taking warfarin. The association of warfarin with bleeding was statistically significant (relative risk 6.185). CONCLUSION: Anticoagulation had no effect on the number of significant perioperative (choroidal) haemorrhages. Aspirin had little effect on bleeding during vitreoretinal surgery. Warfarin, however, was associated with bleeding complications. We suggest that aspirin should not be stopped prior to surgery. Warfarin may be stopped if the patient's thromboembolic risk is low. PMID- 12631018 TI - Vigabatrin-associated visual field constriction in a longitudinal series. Reversibility suggested after drug withdrawal. AB - PURPOSE: To evaluate through a longitudinal study the effects on visual fields of long-term vigabatrin medication in patients with partial epilepsy and to discuss visual field screening strategies. METHODS: A total of 26 patients aged 14-68 years with a mean history of vigabatrin medication of 8.5 years (range 2-14 years) were followed by manual kinetic Goldmann perimetry (objects IV,4 and I,4) for 6-26 months (mean value 12.3 months). At time zero and at follow-up, each patient was assigned a "pooled" averaged value, as a linear percentage of normal isopter position, for the two objects as tested nasally and temporally in the five most horizontal meridians on the Goldmann chart. Twelve eyes from nine adults (age 24-60 years) served as controls. RESULTS: Constrictions were recorded in 24 of 26 patients at baseline. Averaged isopters ranged from 8% to 96% of the controls' averaged isopter positions. Median values of 71.5% and 60.5% for large and small objects, respectively, indicated that the smaller object was more sensitive to visual field constriction. There was no difference in the degree of constriction between nasal and temporal hemifields. Significant improvement in the visual field (mean gain 13.6% units) was seen in the eight patients who underwent full drug withdrawal. No similar improvement was seen in the 12 patients still on full dose or the six with reduced intake. CONCLUSIONS: Most Danish patients on long-term vigabatrin medication have suffered some visual field loss. Contrary to most clinical evidence so far, the present follow-up study indicates some reversibility of visual field loss after drug withdrawal. Kinetic Goldmann perimetry appears to be a fair alternative to computerized static perimetry techniques for screening and following vigabatrin-treated patients. PMID- 12631019 TI - Metastases in the ophthalmic region in Denmark 1969-98. A histopathological study. AB - PURPOSE: To evaluate the patterns of origin and anatomical localization of histologically verified metastases in the ophthalmic region in Denmark during 1969-98, and to compare the findings with those of a similar study for the period 1944-68. METHODS: All metastatic lesions presented at the Eye Pathology Institute, University of Copenhagen during the period 1968-98 were investigated. RESULTS: Metastases were found in 81 specimens from the eye and eye region in 76 patients (32 men, 44 women). Five patients had bilateral involvement. The most frequent site of involvement was the uvea (63%), followed by the orbit (26%) and the eyelids (10%). Breast cancer accounted for the most frequent primary tumour (38%), followed by lung cancer (24%) and skin melanoma (14%). CONCLUSION: The pattern of anatomical location of metastases in the ocular region has not changed since 1944. The incidence of metastases has fallen significantly throughout the period. PMID- 12631020 TI - Detection of connective tissue growth factor (CTGF) in human tear fluid: preliminary results. AB - PURPOSE: Connective tissue growth factor (CTGF) is one of the main regulators of fibrosis. We aimed to evaluate its presence in the human tear fluid of healthy individuals. METHODS: A total of 70 tear fluid samples were collected from eight volunteers prior to and after stimulation of reflex tears with onion vapour. Specific ELISA analysis was performed with goat IgG against human CTGF. RESULTS: Connective tissue growth factor was detected in seven samples (10%), with maximum levels of 17 ng/mL in basal tears. Induction of reflex tearing resulted in a fast and significant decrease of CTGF concentrations (r = - 0.95). No CTGF was detected in 90% of the samples. CONCLUSION: Connective tissue growth factor may occur in tear fluid in healthy human eyes. This indicates a possible role for tear fluid CTGF in ocular surface fibrosis and wound healing. PMID- 12631021 TI - Influence of temperature on corneas stored in culture medium. A comparative study using functional and morphological methods. AB - PURPOSE: To investigate the influence of storage temperature on corneal swelling and on endothelial morphology in cultured corneas. MATERIAL AND METHODS: Forty eight rabbit corneas were separated into four groups of 12. The corneas were stored in culture medium at 37 degrees (group 37), 34 degrees (group 34), 31 degrees (group 31) and 23 degrees (room temperature) (group 23), respectively. All the corneas were monitored by weight recordings on days 0, 1, 2, 3, 4 and 7. On day 7, corneas were prepared for scanning electron microscopy and endothelial cell counts were performed. RESULTS: Lowering the temperature of the culture medium resulted in less swelling. Both temperature and storage time had significant effects on corneal swelling (p < 0.001). On day 7, the observed mean weight increase was 131.2%, 143.0%, 172.5% and 199.7% in groups 23, 31, 34 and 37, respectively. The estimated mean daily weight increase for the corneas were 2.6%, 4.0%, 9.1% and 16.0% in groups 23, 31, 34 and 37, respectively. Scanning electron microscopy showed an intact endothelial layer in all groups after 7 days and there were no statistically significant differences in endothelial counts between groups 23, 31 and 34. In group 37, the cell borders were difficult to distinguish after 7 days and no meaningful count could be performed. CONCLUSIONS: The swelling rate of cultured corneas is significantly less at 23 degrees and 31 degrees than it is at 34 degrees and 37 degrees during the first week. This is most likely the result of a greatly increased barrier effect at lower temperatures. Whereas weight recording revealed profound differences between the groups, scanning electron microscopy and endothelial cell counting did not. The results support the hypothesis that storage at 37 degrees is not optimal in culturing corneas. Lowering the temperature below body temperature, and even lower than 31 degrees, results in less corneal swelling. PMID- 12631022 TI - Indocyanine green angiography in pigmented paravenous retinochoroidal atrophy. AB - PURPOSE: To report long-term follow-up of patients with pigmented paravenous retinochoroidal atrophy (PPRCA) and to assess the involvement of the choroid and retinal pigment epithelium (RPE) in PPRCA. METHODS: Clinical features of PPRCA were studied retrospectively in four patients followed for 6-26 years. Retinal pigment epithelium and choroidal changes were analysed with fluorescein and indocyanine green (ICG) angiography. RESULTS: The two younger patients, aged 16 and 28 years and followed for 6 and 18 years, respectively, showed stationary RPE atrophy and pigmentation. Indocyanine green angiography visualized slight to modest atrophy of the choriocapillaris. The two older patients, aged 69 and 70 years and each followed for 26 years, showed slow progression of disease during follow-up. Indocyanine green angiography revealed choriocapillaris atrophy partly extending into the areas shown as hyperfluorescent in fluorescein angiography. CONCLUSION: Pigmented paravenous retinochoroidal atrophy is probably a slowly progressive disease, particularly in older patients. The choriocapillaris atrophy in this disease is more properly evaluated by ICG angiography and can be underestimated by fluorescein anigiography. PMID- 12631023 TI - Amniotic membrane transplantation elicits goblet cell repopulation after conjunctival reconstruction in a case of severe ocular cicatricial pemphigoid. AB - PURPOSE: The surgical procedures used until now to rebuild the conjunctiva in patients affected by ocular cicatricial pemphigoid (OCP) have not demonstrated appreciable anatomical and functional success. This study reports the postoperative clinical and cytological outcomes of a human amniotic membrane transplantation (AMT) to rebuild the conjunctiva in a case of late stage OCP. METHODS: We present a 75-year-old woman with a severe form of pemphigoid with entropion, trichiasis, symblepharon and blunting of the fornices, who underwent excision of the scarred and inflamed tissue covering the ocular surface and AMT. RESULTS: Our data show an improvement of the ocular surface condition, with reacquired fornix depth, reduced inflammation and presence of goblet cells at each follow-up. CONCLUSION: Amniotic membrane transplantation was successful as a first step procedure to reduce inflammation and to rebuild a physiological conjunctival epithelium in late stage OCP. PMID- 12631024 TI - Indocyanine green angiography in a case of unilateral recurrent posterior acute multifocal placoid pigment epitheliopathy. AB - PURPOSE: To report a case of unilateral recurrent acute posterior multifocal placoid pigment epitheliopathy (APMPPE) imaged by indocyanine green (ICG) angiography. METHODS: A complete ophthalmological evaluation, including fluorescein and ICG angiography, was performed. RESULTS: Acute posterior multifocal placoid pigment epitheliopathy recurred 3 months after the initial disease in a 23-year-old male patient. On ICG angiography, both new, active and healed, inactive lesions demonstrated hypofluorescence throughout the study. The healed lesions were smaller in size and irregularly shaped, whereas the new, active lesions were larger in size, round and more confluent. CONCLUSION: New, active and healed, inactive lesions in APMPPE can both be imaged and differentiated by ICG angiography. PMID- 12631025 TI - Up-irrigation of dropped nuclear fragments during phacoemulsification with the bimanual irrigation-aspiration system. PMID- 12631026 TI - Bilateral optic neuritis in a 9-year-old girl. PMID- 12631027 TI - Late opacification of a hydrophilic acrylic intraocular lens. PMID- 12631028 TI - A long-standing, deep, intracorneal foreign body. PMID- 12631029 TI - Hand dominance, eye laterality and refraction. PMID- 12631030 TI - Recovery of vision after chiasmal optic neuritis. PMID- 12631031 TI - Cyst formation after parotid duct transplantation for severe dry eye. PMID- 12631032 TI - Internet-based counselling of remote ophthalmological patients. PMID- 12631033 TI - Bilateral superior ophthalmic vein thrombosis in a young woman. PMID- 12631034 TI - Paediatric periocular granuloma annulare. PMID- 12631039 TI - New and active role of the interstitium in control of interstitial fluid pressure: potential therapeutic consequences. AB - Here we present recent data indicating that the present view of the interstitium as a passive fluid reservoir has to be revised. The connective tissue cells and extracellular matrix have a role in the control of P(if) and a fundamental role in the rapid development of edema in burns and in the initial swelling in inflammation by generating a lowering of interstitial fluid pressure. In this process, the beta1-integrin system seems to provide a common pathway by which the cells can lower as well as raise P(if). Inflammatory swelling can be reversed by endo- and exogenous substances, thereby suggesting that the connective tissue can serve as a novel target for pharmacological intervention. Furthermore, the new knowledge in interstitial physiology on means to reduce interstitial fluid pressure may be of importance for drug delivery into solid tumors, where a high P(if) limits the uptake of therapeutic agents. PMID- 12631040 TI - Mathematical model of renal elimination of fluid and small ions during hyper- and hypovolemic conditions. AB - This study is concerned with the formulation of a 'kidney module' linked to the plasma compartment of a larger mathematical model previously developed. Combined, these models can be used to predict, amongst other things, fluid and small ion excretion rates by the kidney; information that should prove useful in evaluating values and trends related to whole-body fluid balance for different clinical conditions to establish fluid administration protocols and for educational purposes. The renal module assumes first-order, negative-feedback responses of the kidney to changes in plasma volume and/or plasma sodium content from their normal physiological set points. Direct hormonal influences are not explicitly formulated in this empiric model. The model also considers that the renal excretion rates of small ions other than sodium are proportional to the excretion rate of sodium. As part of the model development two aspects are emphasized (1): the estimation of parameters related to the renal elimination of fluid and small ions, and (2) model validation via comparisons between the model predictions and selected experimental data. For validation, model predictions of the renal dynamics are compared with new experimental data for two cases: plasma overload resulting from external fluid infusion (e.g. infusions of iso-osmolar solutions and/or hypertonic/hyperoncotic saline solutions), and untreated hypo volemic conditions that result from the external loss of blood. The present study demonstrates that the empiric kidney module presented above can provide good short-term predictions with respect to all renal outputs considered here. Physiological implications of the model are also presented. PMID- 12631042 TI - Anesthesiologists in prehospital care make a difference to certain groups of patients. AB - BACKGROUND: Knowledge of the population using prehospital emergency services is scarce except for selected subgroups. Interventions are often made without evaluation. The aim of this study was (1) to describe mortality, hospitalization and the diagnostic pattern among emergency ambulance users and (2) to evaluate the impact of one mobile emergency care unit (MECU) staffed by an anesthesiologist. DESIGN AND METHODS: A descriptive and quasi-experimental study of consecutive emergency ambulance users during two 3-month periods: before the MECU (Period 1) and after (Period 2). Hospitalization, diagnostic and 0-180-day mortality data were requested from national registers. Diagnoses were according to the International Classification of Diseases (ICD). RESULTS: Periods 1 and 2 included 2950 and 2869 users, respectively. The MECU attended 27.7% in Period 2. Fewer users were brought to hospital in Period 2 (87.9% vs. 93.8%, P < 0.0001), especially MECU users (76.5% MECU users vs. 92.3% other users; P < 0.001). Diagnoses included all main ICD-groups. Overall mortality of all users was 10.2%; no difference between the periods. Cardiovascular and respiratory diseases were among the most frequent and were associated with high mortality. In Period 2 mortality was lower in subgroups: acute myocardial infarction (AMI; n = 177, day 0-180, 13.3% vs. 40.5%, P < 0.001); and respiratory diseases, only short-term mortality (n = 388, day 0-1 mortality, 0.0% vs. 2.4%, P < 0.05). CONCLUSION: The diagnostic pattern among emergency ambulance users included all main groups of diseases. After the MECU fewer were brought to hospital. The overall mortality for all ambulance users was not influenced by the MECU. For the subgroups, especially AMI, mortality was lower after the introduction of the MECU. PMID- 12631041 TI - Analgesic effect of i.v. paracetamol: possible ceiling effect of paracetamol in postoperative pain. AB - BACKGROUND: Despite the widespread use of paracetamol for many years, the analgesic serum concentrations of paracetamol are unknown. Therefore the correlation between serum paracetamol concentrations and the analgesic effect was studied. METHODS: Sixty-four women undergoing laparoscopic sterilization were included in a double-blind, placebo-controlled, randomized study. Patients were given i.v. propacetamol 40 mg kg(-1) (group H), 20 mg kg(-1) (group I), 10 mg kg( 1) (group L) or placebo after surgery. Alfentanil was available via patient controlled analgesia (PCA) during the 4-h postoperative study period. The patients' self-reported pain was registered on the visual analog scale (VAS). A pharmacokinetic model was fitted to the paracetamol data. RESULTS: One to 3 h after injection of propacetamol the alfentanil consumption was significantly (P = 0.01-0.04) higher in the placebo group compared with groups H, I, and L receiving propacetamol. There were no significant differences between the amounts of alfentanil consumed in groups H, I, and L. Initial VAS-scores were moderate (5.4 6.2), and declined significantly (P < 0.0001) over time, with no difference between groups. Paracetamol followed an open two-compartment model with i.v. administration and first order elimination. The estimated concentrations immediately (t = 0) after injection were 56 mg l(-1) (H), 28 mg l(-1) (I) and 14 mg l(-1) (L). CONCLUSION: We showed a significant opioid-sparing effect of paracetamol in the immediate postoperative period. Pharmacokinetic data were in accordance with other studies. Our results suggest that a ceiling effect of paracetamol may be present at i.v. doses of 5 mg kg(-1), i.e. a serum concentration of 14 mg l(-1), which is a lower dose than previously suggested. PMID- 12631043 TI - Death after injury in rural Norway: high rate of mortality and prehospital death. AB - BACKGROUND: Finnmark is a rural and remote area in Norway with a sparse population and long distances. Trauma-related mortality has been consistently above the Norwegian national average for the last 20 years. Although the causes of death are well established, very little is known about the time and place of death. This information has implications for the organization of emergency services in rural areas. We examined all trauma deaths over a five-year period in order to inform the debate on how best to reduce our above-average mortality rate. METHODS: A retrospective study of all deaths after trauma (ICD-9 E800-E999) during the years 1991-95 using data obtained from the National Registry of Death. RESULTS: Of the 183 cases found, 130 deaths were due to trauma using definitions comparable to similar studies. The mortality rate was 77 per 100,000 inhabitants per year. Death occurred in the prehospital phase in 85% of cases. Seventy-two per cent of all deaths (regardless of location) occurred within the first h after injury, eight per cent from 1 to 4 h and the remaining 20% occurred after 4 h. CONCLUSION: When planning interventions to reduce the mortality rate from trauma in rural areas, a high proportion of prehospital deaths should be expected. The high number of patients who are found dead (which can only be reduced by injury prevention) must be taken into account. Measures to reduce 'preventable' causes of death by bystanders should be evaluated. Further knowledge of exact mechanisms of death in the prehospital phase is required. PMID- 12631044 TI - The difference between sleep and anaesthesia is in the intracellular signal: propofol and GABA use different subtypes of the GABA(A) receptor beta subunit and vary in their interaction with actin. AB - BACKGROUND: Propofol is known to interact with the gamma-aminobutyric acidA (GABA(A)) receptor, however, activating the receptor alone is not sufficient for producing anaesthesia. METHODS: To compare propofol and GABA, their interaction with the GABAA receptor beta subunit and actin were studied in three cellular fractions of cultured rat neurons using Western blot technique. RESULTS: Propofol tyrosine phosphorylated the GABA(A) receptor beta2 (MW 54 and 56 kDa) and beta3 (MW 57 kDa) subtypes. The increase was shown in both the cytoskeleton (beta2(54) and beta2(56) subtypes) and the cell membrane (beta2(54) and beta3 subtypes). Concurrently the 56 kDa beta2 subtype was reduced in the cytosol. Propofol, but not GABA, also tyrosine phosphorylated actin in the cell membrane and cytoskeletal fraction. Without extracellular calcium available, the amount of actin decreased in the cytoskeleton, but tyrosine phosphorylation was unchanged. GABA caused increased tyrosine phosphorylation of beta2(56) and beta3 subtypes in the membrane and both beta2 subtypes in the cytoskeleton but no cytosolic tyrosine phosphorylation. CONCLUSION: The difference between propofol and GABA at the GABA(A) receptor was shown to take place in the membrane, where the beta2(54) was increased by propofol and instead the beta2(56) subtype was increased by GABA. Only propofol also tyrosine phosphorylated actin in the cell membrane and cytoskeletal fraction. This interaction between the GABAA receptor and actin might explain the difference between anaesthesia and physiological neuronal inhibition. PMID- 12631045 TI - Different benefit of bispectal index (BIS) in desflurane and propofol anesthesia. AB - BACKGROUND: Bispectal index (BIS) monitoring may reduce drug usage and hasten recovery in propofol and inhalation anesthesia. The faster emergence profile of desflurane may reduce the effect of BIS monitoring on recovery from desflurane compared with propofol. This study compared hypnotic drug usage, recovery, patient satisfaction and incidence of inadequate sedation in BIS monitored and nonmonitored women anesthetized with desflurane or propofol. METHODS: One hundred and sixty patients scheduled for elective gynecological surgery were randomly assigned to desflurane or propofol anesthesia with and without BIS monitoring. Fentanyl, vecuronium and remifentanil were administered according to clinical criteria. The BIS monitor was used in all patients, but the monitor screen was covered in the controls. A BIS level between 45 and 55 was targeted in the BIS monitored patients whereas depth of anesthesia was assessed by clinical criteria in the controls. RESULTS: The mean (SD) desflurane MAC-hours administered with and without BIS were 0.70 (0.15) and 0.76 (0.12), respectively, resulting in extubation times of 6.5 (4.1) and 8.3 (6.1) min. (NS). Bispectal index monitoring was associated with improved patient satisfaction, reduced postoperative nausea and antiemetic drug requirement, and fewer episodes with sustained BIS levels > 60. The mean (SD) propofol infusion rates were 6.0 (1.4) and 6.6 (0.9) mg kg( 1)h(-1) with and without the BIS monitor (P = 0.023), resulting in mean (SD) extubation times of 6.8 (4.6) and 10.5 min (5.9), respectively (P < 0.05). CONCLUSION: Bispectal index monitoring reduced propofol usage and hastened recovery after propofol anesthesia, whereas in desflurane anesthesia it was associated with improved patient satisfaction, probably because of decreased postoperative nausea and fewer episodes of inadequate hypnosis. PMID- 12631047 TI - Effects of high-dose propofol on succinylcholine-induced fasciculations and myalgia. AB - BACKGROUND: The purpose of this prospective study was to determine the effects of high-dose propofol on the incidence of fasciculations and myalgia, and to evaluate changes in creatine kinase levels following the administration of succinylcholine in 90 women who underwent laparoscopy. METHODS: Patients were randomly assigned to one of three groups. Induction of anesthesia was performed with thiopentone 5 mg kg(-1) in Group I (n = 30), propofol 2 mg kg(-1) in Group II (n = 30), and propofol 3.5 mg kg(-1) in Group III (n = 30). Then succinylcholine 1 mg kg(-1) was administered to the patients for intubation. RESULTS: Fasciculation was absent in 20% of Group III patients, and no vigorous fasciculation occurred in this group. Furthermore, the severity of fasciculation in Group III was significantly lower than in the other two groups (P = 0.01). Seventy per cent of patients had no myalgia in Group III, 39.2% in Group II and 37% in Group I (P = 0.007). Severity of myalgia was also significantly lower in Group III compared with the other two groups (P = 0.011). Post-operative creatine kinase levels were significantly higher than their baseline values in Groups I and II (P < 0.0001). CONCLUSION: Administration of propofol 3.5 mg kg-1 is effective in reducing fasciculations and myalgia after succinylcholine. PMID- 12631046 TI - Sevoflurane versus isoflurane--anaesthesia for lower abdominal surgery. Effects on perioperative glucose metabolism. AB - BACKGROUND: The aim of this study was to determine the impact of sevoflurane anaesthesia on metabolic and endocrine responses to lower abdominal surgery. METHODS: A prospective randomized controlled study in 20 patients undergoing abdominal hysterectomy. Patients were randomly assigned to receive either sevoflurane (S) or isoflurane anaesthesia (I). Using a stable isotope dilution technique, endogenous glucose production (EGP) and plasma glucose clearance (GC) were determined pre- and postoperatively (6,6-2H2-glucose). Plasma concentrations of glucose, insulin, cortisol, epinephrine and norepinephrine were measured preoperatively, 5 min after induction of anaesthesia, during surgery and 2 h after the operation. RESULTS: EGP increased in both groups with no intergroup differences (preop. S 12.2 +/- 1.6, I 12.4 +/- 1.6; postop. S 16.3 +/- 1.9*, I 19.0 +/- 3.1* micromol kg(-1) min(-1), all values are means +/- SD, *P < 0.05 vs. preop.). Plasma glucose concentration increased and GC decreased in both groups. There were no differences between groups. (Glucose conc. mmol l(-1) preop.: S 4.1 +/- 0.3, I 3.9 +/- 0.5; 5 AI S 5.1 +/- 0.6*, I 5.1 +/- 1.0*, postop. S 7.0 +/- 1.0*, I 7.1 +/- 1.4*; * = P < 0.05 vs. preop.; GC ml kg(-1)min(-1) preop. S 3.0 +/- 0.4, I 3.2 +/- 0.4; postop. S 2.4 +/- 0.3*, I 2.7 +/- 0.3*; *=P < 0.05 vs. preop.) Insulin plasma concentrations were unchanged. Cortisol plasma concentrations increased intra- and postoperatively with no changes between the groups. Norepinephrine plasma concentration increased in the S group after induction of anaesthesia. I group norepinephrine was increased 2 h after operation and showed no intergroup differences. CONCLUSION: Sevoflurane, as well as isoflurane, does not prevent the metabolic endocrine responses to surgery. PMID- 12631048 TI - Oral clonidine premedication preserves heart rate variability for patients undergoing larparoscopic cholecystectomy. AB - BACKGROUND: Clonidine has been shown to reduce perioperative circulatory instability. This postoperative analgesic effect of clonidine was also known in previous studies. The aim of the study was to investigate the clinical efficiency of oral clonidine premedication in anesthesia and analgesia in patients undergoing laparoscopic cholecystectomy. METHODS: Thirty-two patients scheduled for elective laparoscopic cholecystectomy were recruited for a prospective, randomized, double-blinded comparative study. They were allotted randomly to two groups: placebo or clonidine. Patients in the placebo group (n = 16) were premedicated with oral antacid (alugel hydroxide 300 mg), while those in the clonidine group (n = 16) were premedicated with oral clonidine 150 micro g before anesthesia. Analysis of heart rate variability was used to quantify the control of heart rate at baseline, and during the pneumoperitoneum and recovery periods. Time of the first request for postoperative analgesic and cumulative analgesic requirements in 24 h were recorded. Data are expressed as mean +/- SD. RESULTS: Heart rate variability was reduced in the pneumoperitoneum and postoperative period in the placebo group. Clonidine resulted in a greater perioperative (pneumoperitoneum period) power at all frequency ranges compared with placebo (671.5 +/- 470.5 vs. 55.1 +/- 51.6 ms2/Hz for total power variability, 170.1 +/- 94.4 vs. 16.9 +/- 21.1 ms2/Hz for low-frequency variability and 206.1 +/- 95.7 vs. 16.4 +/- 15.1 ms2/Hz for high-frequency variability, P < 0.05). The postoperative analgesic requirement was less (2.3 +/- 0.8 vs. 3.2 +/- 1.2 dose, P < 0.05) in comparison with the placebo group. CONCLUSION: Clonidine preserves heart rate control in pneumoperitoneum and recovery periods. Oral clonidine premedication also reduces the requirement for postoperative analgesia. PMID- 12631049 TI - Effects of preoperative oral carbohydrates and peptides on postoperative endocrine response, mobilization, nutrition and muscle function in abdominal surgery. AB - BACKGROUND: Surgery is succeeded by long-lasting state of relative peripheral insulin resistance, which is reduced by giving glucose infusion or oral carbohydrate-rich drinks immediate before operating instead of fasting. The aim of the present study was to investigate whether oral carbohydrate or carbohydrate with peptide drinks preoperatively instead of fasting would improve postoperative voluntary muscle strength, nutritional intake and ambulation, decrease postoperative fatigue, anxiety and discomfort, and reduce the endocrine response to surgery. METHODS: Forty-eight patients were included and randomized into three groups to receive 2 x 400 ml of carbohydrate-rich drinks or to fast overnight and allowed only water. Voluntary grip and quadriceps strength, body composition, pulmonary function, VAS-score of eight parameters of wellbeing, muscle biopsies and insulin, glucagon, IGF-1 and free fatty acids were measured before and after the operation. The basic postoperative regimen for all groups were immediate oral nutrition and early enforced mobilization. RESULTS: Significant postoperative decrease in glycogen synthase activity in the muscle biopsies was reduced in the intervention groups, and in combination, the intervention groups had a less reduced quadriceps strength after one week (-10% vs. -16%, NS) and one month (-5% vs. -13%, P < 0.05). Minor changes in the endocrine response to surgery were found without differences between the groups, and there were no differences between the groups in ambulation time, nutritional intake or subjective measures of wellbeing. PMID- 12631051 TI - Preoperative blood pressure and catecholamines related to hypothermia during general anesthesia. AB - BACKGROUND: We previously demonstrated that preoperative blood pressure values affect intraoperative hypothermia during general anesthesia. We hypothesized that increased catecholamine secretion could be responsible for the relationship between preoperative blood pressure and hypothermia. METHODS: To evaluate the effect of preoperative systolic blood pressure (SBP) and plasma catecholamine levels on core temperature during general anesthesia, 40 male patients who were scheduled for open abdominal surgery were allocated to two groups: those whose preoperative SBP was 140 mmHg or greater (high SBP group, n = 20), and those whose SBP was less than 140 mmHg (normal SBP group, n = 20). Anesthesia was maintained with 0.4% isoflurane and opioids. RESULTS: The average age, height, and weight of the patients in the two groups did not differ. Preoperative SBP, mean blood pressure, diastolic blood pressure and heart rate in the high SBP group were significantly higher than those in the normal SBP group. Plasma norepinephrine concentrations in the high SBP group were significantly greater than those in the normal SBP group before and 1 h after the induction of anesthesia. Tympanic membrane temperatures in the normal SBP group started to decline further just after the induction of anesthesia, more so than that in the high SBP group. The vasoconstriction threshold in the normal SBP group was significantly lower than that in the high SBP group. CONCLUSION: These results suggest that the higher levels of preoperative catecholamine secretion contributed to the lesser degree of intraoperative hypothermia observed in the high SBP group. PMID- 12631052 TI - Effects of human cathelicidin antimicrobial peptide LL-37 on lipopolysaccharide induced nitric oxide release from rat aorta in vitro. AB - BACKGROUND: Lipopolysaccharides (LPS), released by Gram-negative bacteria, cause vascular expression of inducible nitric oxide synthase (iNOS) leading to nitric oxide (NO) production and septic shock. Human cathelicidin antimicrobial peptide (LL-37) can bind and neutralize LPS. We wanted to study whether LL-37 affects LPS or interleukin-1beta (IL-1beta)-induced production, release and function of NO in intact rat aorta rings and cultured rat aorta smooth muscle cells. METHODS: Isolated segments of thoracic aorta and cultured cells were incubated in the presence of LPS, LL-37, LPS + IL-37, IL-1beta, IL-1beta + IL-37 or in medium alone. Smooth muscle contraction in response to phenylephrine and accumulation of the sdegradation products of NO, nitrate and nitrite, were measured on aorta segments. Levels of iNOS were assessed by Western blot and cytotoxic effects were detected by measurement of DNA fragmentation in cultured cells. Number of viable cells were determined after Trypan blue treatment. RESULTS: Both LPS and IL-1beta reduced contractility in response to phenylephrine and increased NO production as well as iNOS expression. LL-37 inhibited the LPS depression of vascular contractility induced only by LPS. LL-37 reduced both the LPS- and IL-1beta induced NO production and iNOS expression. LL-37 at high concentrations induced DNA fragmentation and decreased the number of living cells. CONCLUSION: IL-37 reduces NO production induced by LPS and IL-1beta. The reduction does not seem to result only from neutralization of LPS but also from a cytotoxic effect, possibly via induction of apoptosis. PMID- 12631050 TI - No influence of the endothelin receptor antagonist bosentan on basal and indomethacin-induced reduction of cerebral blood flow in pigs. AB - BACKGROUND: The mechanism behind indomethacin-induced cerebral vasoconstriction is incompletely understood. We tested the hypothesis that the mixed endothelin-1 receptor antagonist bosentan would modify or prevent indomethacin-induced reduction of CBF in the anaesthetized pig. Furthermore, we investigated the effect of bosentan on resting CBF and CMRO2. METHODS: Twelve pigs were randomized in two groups of six, and received either bosentan and indomethacin (group 1), or placebo and indomethacin (group 2). Anaesthesia was induced with ketamine and midazolam and maintained with fentanyl, nitrous oxide and pancuronium. Baseline measurements of CBF and CMRO2 were performed before intravenous bolus injection of bosentan (10 mg/kg) or placebo (0.9% NaCl). The second CBF and CMRO2 measurement was performed 30 min after administration of bosentan/placebo. A 40 min infusion of indomethacin (0.05 mg/kg/min) was administered and the third CBF and CMRO2 measurement was performed 80 min after administration of bosentan/placebo. Independently, pharmacokinetic data of bosentan were generated in four pigs. RESULTS: In group 1, baseline CBF was 55 +/- 7 ml/100 cm3/min. Administration of bosentan i.v. did not change CBF significantly. Indomethacin decreased CBF to 41 +/- 5 ml/100 cm3/min (P < 0.002). In group 2, baseline CBF was 54 +/- 10 ml/100 cm3/min. Placebo did not change CBF while indomethacin decreased CBF significantly to 41 +/- 5 ml/100 cm3/min (P < 0.002). No significant changes in CMRO2 were observed. In group 2, a significant increase in MABP was observed after administration of indomethacin. No change in MABP was observed in the bosentan-treated animals. Total plasma concentrations of bosentan at the time of the first and the second PET measurement were 3.9 and 1.4 microg/ml, respectively. The corresponding values for the pharmacologically active metabolite Ro 48-5033 were 1.2 and 0.4 microg/ml. CONCLUSION: These findings indicate that endothelin receptor stimulation is not involved in indomethacin-induced cerebral vasoconstriction or maintenance of cerebrovascular tone in the anaesthetized pig. However, our results suggest that the increase in MABP is mediated through endothelin receptors. PMID- 12631053 TI - Glyburide decreases myocardial oxygen pressure in dogs. AB - BACKGROUND: Reports show that glyburide, an adenosine triphosphate sensitive potassium (K+ATP) channel blocker, will reverse the myocardial protective effect of inhalational anesthesia. We evaluated the effect of glyburide on myocardial tissue oxygen pressure (PmO2) in dogs anesthetized with desflurane. METHODS: Twelve dogs were anesthetized with 8% end-tidal desflurane for baseline anesthesia. A flow probe was placed on the left anterior descending (LAD) artery. A probe that measured PmO2 was inserted into the middle myocardium in the LAD region. After baseline measures, six dogs received i.v. 1 mg kg(-1) of glyburide and six dogs received sham vehicle treatment. After the glyburide or sham treatment, each dog received an i.v. infusion of adenosine 0.1 microg kg(-1) x min(-1), sodium nitroprusside (SNP) 2-4 microg kg(-1) x min(-1) and 14% end-tidal desflurane in random order. RESULTS: Glyburide decreased LAD artery flow from 59 +/- 9 ml min(-1) to 30 +/- 6 ml min(-1) (P < 0.05) and PmO2 from 44 +/- 16 mmHg to 30 +/- 9 mmHg (P < 0.05). Adenosine infusion increased LAD artery blood flow 180% in the sham-treated dogs but produced no change in the glyburide-treated dogs. Sodium nitroprusside infusion increased LAD artery flow and decreased PmO2 in both the glyburide- and sham-treated dogs. Desflurane (14%) did not reverse the glyburide-induced vasoconstriction but increased PmO2 to 38 +/- 20 mmHg (P < 0.05). CONCLUSION: Glyburide produced myocardial tissue hypoxia, which was not changed by adenosine, worsened by SNP and improved by 14% desflurane. The improvement in PmO2 with desflurane occurred without a change in myocardial blood flow. PMID- 12631054 TI - Exotic snake bite: a challenge for the Scandinavian anesthesiologist? AB - BACKGROUND: Venomous snake bites are uncommon in the Scandinavian countries. Envenomation from exotic snakes do however occur, mostly amongst snake handlers. This case report documents the effects and treatment for envenomation from Hoplocephalus bungaroides, or the Broad-Headed snake, native to eastern and southern Australia. Snakes of the genus Hoplocephalus have previously been described as of 'lesser medical importance' because of their rarity. METHODS: This case report describes the signs, symptoms and management of systemic envenomation in a previously healthy man. RESULTS: The patient developed signs of severe coagulopathy less than an hour after envenomation. There was also biochemical evidence of rhabdomyolysis, and cardiotoxicity. At no time did the patient develop respiratory insufficiency, neurotoxicity or renal failure. The patient was initially managed with i.v. crystalloids, plasma, corticosteroids and antifibrinolytics and by observation in the intensive care unit (ICU). Coagulopathy resolved after causal treatment with monovalent Tiger snake antivenom. CONCLUSION: The patient made good progress and was well on discharge from the ICU 26 h postenvenomation. PMID- 12631055 TI - Complete recovery from prolonged cardio-pulmonary resuscitation following anaphylactic reaction to readministered intravenous cefazolin. AB - We describe a patient who developed a type I anaphylactic reaction to intravenous cefazolin. The patient had no known drug allergies and had previously received intraoperative intravenous cefazolin 2 months prior without any problems. Forty fives after receiving cefazolin 1 g i.v. and while fully awake, the patient experienced shortness of breath, became unconscious, and then suffered a cardiac arrest. Resuscitation included endotracheal intubation, external cardiac compression, electrical defibrillation and multiple large doses of epinephrine, atropine, and sodium bicarbonate over the course of 2.5 h and three cardiac arrests. Nevertheless, the patient fully recovered. The intent of this case report is to address widely held concerns regarding cross-reactivity of cephalosporin, particularly cefazolin, to penicillin, the legitimacy of test dosing as a means to safely identify patients who will have an allergic reaction to cephalosporins and comment on patient-related predictors of survival following cardiopulmonary resuscitation and the good outcome in this case. PMID- 12631056 TI - Diagnosis of malignant hyperthermia susceptibility during CABG surgery. AB - A 55-year-old man presented for coronary artery bypass graft (CABG) surgery. Malignant hyperthermia (MH) was suspected in his family. This case report describes a diagnostic approach to obtain a definite MH diagnosis by performing an in vitro contracture test at the time of CABG surgery in combination with molecular genetic investigations. PMID- 12631057 TI - Loss of vision after spinal anesthesia. PMID- 12631058 TI - Body art, the parturient and obstetrical anesthesia. PMID- 12631059 TI - Procalcitonin as a marker. PMID- 12631061 TI - Reverse epidemiology of cardiovascular risk factors in maintenance dialysis patients. AB - Conventional risk factors of cardiovascular disease and mortality in the general population such as body mass, serum cholesterol, and blood pressure are also found to relate to outcome in maintenance dialysis patients, but often in an opposite direction. Obesity, hypercholesterolemia, and hypertension appear to be protective features that are associated with a greater survival among dialysis patients. A similar protective role has been described for high serum creatinine and possibly homocysteine levels in end-stage renal disease (ESRD) patients. These findings are in contrast to the well-known association between over nutrition and poor outcome in the general population. The association between under-nutrition and adverse cardiovascular outcome in dialysis patients, which stands in contrast to that seen in non-ESRD individuals, has been referred to as "reverse epidemiology." Publication bias may have handicapped or delayed additional reports with such paradoxical findings in ESRD patients. The etiology of this inverse association between conventional risk factors and clinical outcome in dialysis patients is not clear. Several possible causes are hypothesized. First, survival bias may play a role since only a small number of patients with chronic kidney disease (CKD) survive long enough to reach ESRD. Hence, the dialysis patients are probably a distinctively selected population out of CKD patients and may not represent the risk factor constellations of their CKD predecessors. Second, the time discrepancy between competitive risk factors may play a role. For example, the survival disadvantages of under-nutrition, which is frequently present in dialysis patients, may have a major impact on mortality in a shorter period of time, and this overwhelms the long-term negative effects of over-nutrition on survival. Third, the presence of the "malnutrition-inflammation complex syndrome" (MICS) in dialysis patients may also explain the existence of reverse epidemiology in dialysis patients. Both protein-energy malnutrition and inflammation or the combination of the two are much more common in dialysis patients than in the general population and many elements of MICS, such as low weight-for-height, hypocholesterolemia, or hypocreatininemia, are known risk factors of poor outcome in dialysis patients. The existence of reverse epidemiology may have a bearing on the management of dialysis patients. It is possible that new standards or goals for such traditional risk factors as body mass, serum cholesterol, and blood pressure should be considered for these individuals. PMID- 12631062 TI - Pathophysiology of proteinuria. AB - Proteinuria is consequence of two mechanisms: the abnormal transglomerular passage of proteins due to increased permeability of glomerular capillary wall and their subsequent impaired reabsorption by the epithelial cells of the proximal tubuli. In the various glomerular diseases, the severity of disruption of the structural integrity of the glomerular capillary wall correlates with the area of the glomerular barrier being permeated by "large" pores, permitting the passage in the tubular lumen of high-molecular-weight (HMW) proteins, to which the barrier is normally impermeable. The increased load of such proteins in the tubular lumen leads to the saturation of the reabsorptive mechanism by the tubular cells, and, in the most severe or chronic conditions, to their toxic damage, that favors the increased urinary excretion of all proteins, including low-molecular-weight (LMW) proteins, which are completely reabsorbed in physiologic conditions. Recent clinical studies showed that in patients with glomerular diseases the urinary excretion of some HMW proteins [immunoglobulins G and M (IgG and IgM)] and of some LMW proteins, alpha1-microglobulin, beta2 microglobulin, correlates with the severity of the histologic lesions, and may predict, better than the quantity of proteinuria, the natural course, the outcome, and the response to treatment. It is suggested that some patients have already, at the time of clinical presentation, a structural damage of the glomerular capillary wall (injury of podocytes) and of the tubulointerstitium, the severity and scarce reversibility of which are reliably indicated by an elevated urinary excretion of HMW and LMW proteins. PMID- 12631063 TI - Laminin-1 reexpression in Alport mouse glomerular basement membranes. AB - BACKGROUND: Alport disease is a heritable basement membrane disorder caused by mutations in genes encoding the alpha3, alpha4, or alpha5 chains of type IV collagen, which normally comprise the collagenous network of mature glomerular basement membranes (GBMs). In Alport disease, the alpha3(IV), alpha4(IV), alpha5(IV) collagen network is absent and substituted for by alpha1(IV), and alpha2(IV) collagen, which normally is present only in developing, immature GBMs. The disease is marked by progressive GBM thickening and delamination, proteinuria, and renal failure. In addition to collagen IV dysregulation, abnormal GBM laminins also occur and may contribute to the pathogenesis of Alport glomerulopathy. METHODS: To investigate laminin dysregulation in a mouse model of Alport disease, we used antibodies specific for laminin-alpha1 and -beta1 chains (to recognize laminin-1), and -alpha5 chain (to recognize laminin-11), and evaluated their distribution during glomerular development in alpha3(IV) collagen deficient mice. RESULTS: Developing glomeruli of infant alpha3(IV) collagen knockout mice underwent normal down-regulation of laminin-1, but laminin-1 chains were then reexpressed in maturing glomeruli, becoming concentrated in the subepithelial GBM projections typical of Alport disease. Immunoelectron microscopy showed that laminin-1 reexpression took place in both glomerular endothelial cells and podocytes. CONCLUSIONS: The absence of a alpha3(IV), alpha4(IV), alpha5(IV) network may stimulate reexpression of laminin-1 by Alport mouse endothelial cells and podocytes. This abnormal GBM, which is more characteristic of immature glomeruli, may promote podocyte foot process effacement and reversion to a less differentiated state. PMID- 12631064 TI - Evidence that bone morphogenetic protein 4 has multiple biological functions during kidney and urinary tract development. AB - BACKGROUND: We have suggested that bone morphogenetic protein 4 (BMP4), acting on the Wolffian duct and ureter epithelium, determines the budding site of the ureter by locally antagonizing ubiquitous inductive signal(s) from the metanephric mesenchyme. In the present study, we examine the effect of BMP4 on the development of metanephric and periureteral mesenchymal cells, which express the BMP type I receptor gene, Bmpr1a (Alk3). METHODS: Urogenital tissues obtained from Bmp4 heterozygous null mutant (Bmp4+/-) embryos at different stages, and metanephric and ureteral tissue explants cultured in the presence of recombinant BMP4 were subjected to morphologic, immunohistochemical and in situ hybridization analyses. To examine the chemotactic activity of BMP4 for periureteral mesenchymal cells, a modified Boyden chamber assay was performed. RESULTS: Many of the kidneys of newborn Bmp4+/- mice contained multicystic dysplastic regions. This morphology was preceded by abnormally high apoptotic activity in the metanephric mesenchyme of mutant embryos at E14.5. In whole metanephric explants, BMP4 uniformly promoted the expansion of the Pax2-negative and weakly Foxd1 (previously Bf2) -positive peripheral stromal compartment of metanephric mesenchyme in the presence of fibroblast growth factor 2 (FGF2). In addition, in isolated metanephric mesenchyme, BMP4-loaded beads prevented apoptosis locally. Thus, BMP4 prevents cell death and promotes the growth of the metanephric mesenchyme. The effect of BMP4 on periureteral mesenchyme is different from its effect on metanephric mesenchyme. In utero, periureteral mesenchymal cells condense around the ureter epithelium, followed by differentiation into smooth muscle cells at a site where Bmp4 is intensely expressed. Analysis of Bmp4+/- ureters at E15.5 reveals that the alpha-smooth muscle actin (alpha-SMA)-positive cells are low in number. In vitro, BMP4-loaded beads promote the accumulation of periureteral mesenchymal cells to form several cell layers surrounding the beads. In addition, in a Boyden chamber assay, BMP4 increases the migration of periureteral mesenchymal cells through the filter. Thus, BMP4 can serve as a chemoattractant for periureteral mesenchymal cells and induce locally the smooth muscle layer of the ureter at Bmp4-expressing sites. CONCLUSION: Depending on local context, BMP4 has several biological actions on the morphogenesis of different portions of the excretory system, namely, the development of the ureterovesical junction, the ureter, and the kidney. PMID- 12631065 TI - Increased endothelin-1 expression in the kidney in hypercalcemic rats. AB - BACKGROUND: Although hypercalcemia causes diuresis and natriuresis, the molecular mechanisms of these effects are not well established. Recently, the important role of the calcium-sensing receptor (CaR) in hypercalcemia-induced polyuria was reported. Endothelin-1 (ET-1) that is locally produced in the nephron has been suggested to have the natriuretic and/or diuretic effects in the kidney. Therefore, we hypothesized that ET-1 expression could be increased through the activation of CaR in the kidney in hypercalcemia. METHODS: Rats were made hypercalcemic by dihydrotachysterol (DHT) treatment. The urinary concentration of ET-1 and the mRNA expression of ET-1 in the kidney were determined. Immunohistochemistry was performed to determine types of the cells that produce ET-1. CaR and ET-1 promoter luciferase constructs were co-expressed in COS-7 cells and the ET-1 promoter activity following the addition of extracellular calcium was measured by the luciferase assay. RESULTS: In hypercalcemic rat, urinary ET-1 excretion was increased by twofold, and ET-1 mRNA expression was increased in the kidney cortex by threefold. In cortical collecting duct (CCD), both principal cells and intercalated cells synthesized ET-1. In cells that express CaR, ET-1 promoter was activated in a dose-dependent manner by extracellular calcium over the range of 0.5 to 3.0 mmol/L. CONCLUSIONS: First, activation of CaR increases ET-1 transcription in a dose-dependent manner. Second, hypercalcemia increases ET-1 production in the kidney cortex. These data suggest the possibility that CaR might play an important role in hypercalcemia induced increase in ET-1 production. PMID- 12631066 TI - GW274150, a potent and highly selective inhibitor of iNOS, reduces experimental renal ischemia/reperfusion injury. AB - BACKGROUND: Generation of nitric oxide (NO) by inducible nitric oxide synthase (iNOS) may contribute to renal ischemia/reperfusion (I/R) injury. The aim of this study was to investigate the effects of GW274150, a novel, highly selective, potent and long-acting inhibitor of iNOS activity in rat and mouse models of renal I/R. METHODS: Rats were administered GW274150 (5 mg/kg intravenous bolus administered 30 minutes prior to I/R) and subjected to bilateral renal ischemia (45 minutes) followed by reperfusion (6 hours). Serum and urinary indicators of renal dysfunction, tubular and reperfusion injury were measured, specifically, serum urea, creatinine, aspartate aminotransferase (AST) and N-acetyl-beta-d glucosaminidase (NAG) enzymuria. In addition, renal sections were used for histologic scoring of renal injury and for immunologic evidence of nitrotyrosine formation and poly [adenosine diphosphate (ADP)-ribose] (PAR). Nitrate levels were measured in rat plasma using the Griess assay. Mice (wild-type, administered 5 mg/kg GW274150, and iNOS-/-) were subjected to bilateral renal ischemia (30 minutes) followed by reperfusion (24 hours) after which renal dysfunction (serum urea, creatinine), renal myeloperoxidase (MPO) activity and malondialdehyde (MDA) levels were measured. RESULTS: GW274150, administered prior to I/R, significantly reduced serum urea, serum creatinine, AST, and NAG indicating reduction of renal dysfunction and injury caused by I/R. GW274150 reduced histologic evidence of tubular injury and markedly reduced immunohistochemical evidence of nitrotyrosine and PAR formation, indicating reduced peroxynitrite formation and poly (ADP ribose) polymerase (PARP) activation, respectively. GW274150 abolished the rise in the plasma levels of nitrate (indicating reduced NO production). GW274150 also reduced the renal dysfunction in wild-type mice to levels similar to that observed in iNOS-/- mice subjected to I/R. Renal MPO activity and MDA levels were significantly reduced in wild-type mice administered GW274150 and iNOS-/- mice subjected to renal I/R, indicating reduced polymorphonuclear leukocyte (PMN) infiltration and lipid peroxidation. CONCLUSIONS: These results suggest that (1). an enhanced formation of NO by iNOS contributes to the pathophysiology of renal I/R injury and (2). GW274150 reduces I/R injury of the kidney. We propose that selective inhibitors of iNOS activity may be useful against renal dysfunction and injury associated with I/R of the kidney. PMID- 12631067 TI - Calpain is activated in experimental uremia: is calpain a mediator of uremia induced myocardial injury? AB - BACKGROUND: The cysteine proteases calpain and caspase-3 are known mediators of cell death. The aim of this study was to assess their contribution to the tissue damage found in experimental uremia. METHODS: Calpain and caspase-3 activities were measured in the hearts of rats that were sham-operated (control), sham operated and spontaneously hypertensive (SHR), and those rendered uremic by 5/6 nephrectomy (uremic). In an in vitro study, heart myoblasts (Girardi) were incubated with human serum from healthy subjects (control serum conditioned media, CSCM) or uremic patients (uremic serum conditioned media, USCM), in the presence and absence of calpain and caspase-3 inhibitors. After 48 hours the activity of calpain and caspase-3 was measured, and cell injury determined by DNA fragmentation (ELISA) and lactate dehydrogenase (LDH) release. An in situ assay was designed to study how USCM affects calpain activity over time. RESULTS: In the in vivo study, mean calpain activities were almost identical in the control and SHR groups, but calpain and caspase-3 activities were much elevated in the uremic group (P < 0.01 and 0.001 respectively vs. control). The SHR group had significantly higher mean arterial blood pressure (P < 0.001 vs. control, 0.01 vs. uremic). In the in vitro study calpain activity and DNA fragmentation were markedly higher in USCM treated cells compared to CSCM (both P<0.05). Both were reduced in USCM cells containing calpain inhibitors (E64d, calpastatin, or PD 150606). LDH release was raised also in USCM treated cultures (P < 0.05), which only the E64d treatment could significantly reduce (P < 0.02). Caspase-3 activities were similar in USCM and CSCM groups. The in situ assay showed significant increases in calpain activity in USCM treated cells compared to CSCM after just 3.5 hours (P<0.01). CONCLUSIONS: In vivo results suggest that the increases in calpain and caspase-3 activity in uremic rat hearts were primarily due to uremia and not to hypertension. In vitro data demonstrate that uremia induced cell injury can be attenuated by calpain inhibition. Therefore, it is likely that calpain is a mediator of uremia-induced myocardial injury. PMID- 12631068 TI - Glucose-induced fibronectin and collagen type III expression in renal fibroblasts can occur independent of TGF-beta1. AB - BACKGROUND: Various renal cell types have been shown to contribute to the excessive matrix deposition observed in diabetic nephropathy. The present study examined the effect of high ambient glucose and transforming growth factor-beta1 (TGF-beta1) on matrix production by human renal fibroblasts. METHODS: Human renal fibroblasts (TK173) were used to examine the effects of high glucose and TGF beta1 on fibronectin and collagen type III expression. Stable transfectants were generated of TK173 cells expressing a dominant negative TGF-beta type II receptor. Matrix components were measured in enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Fibronectin secretion by renal fibroblasts was increased upon exposure to high glucose, but with delayed kinetics compared to TGF-beta1-induced fibronectin. Exposure to high glucose resulted in an increased secretion of latent TGF-beta1. However, treatment with neutralizing pan-specific anti-TGF-beta antibodies could not attenuate the effects of glucose. Furthermore, collagen type III was up regulated by high glucose, but not by TGF-beta1. Importantly, fibroblasts expressing a dominant negative TGF-beta type II receptor were defective in TGF beta1-induced fibronectin production, whereas glucose-induced fibronectin and collagen type III were unaffected. CONCLUSIONS: These data show that in renal fibroblasts exposure to high glucose can increase matrix production independent of endogenous TGF-beta1. Although glucose activation is accompanied by an increased production of latent TGF-beta1, which can have an important role in vivo, the data suggest involvement of alternative growth factors in the mechanism by which hyperglycemic conditions can modulate matrix accumulation in diabetic nephropathy. PMID- 12631069 TI - Monocyte adhesion to mesangial matrix modulates cytokine and metalloproteinase production. AB - BACKGROUND: Monocytes migrate into the glomerular mesangium during acute inflammatory renal disease, differentiate into macrophages, and may play a key role in the development and progression of glomerular scarring. Treatment strategies that inhibit monocyte infiltration ameliorate glomerular injury in animal models. Mesangial matrix contains several potential monocyte-binding domains that may contribute to monocyte entrapment and modulate cell activation. METHODS: Adhesion of peripheral blood-derived monocytes to matrix synthesized by human mesangial cells and to individual matrix proteins was assessed by colorimetry of nuclear staining with crystal violet. Monoclonal antibodies were used to identify the cell-surface integrins and matrix ligands involved. Monocyte proliferation was assessed by 3H-thymidine incorporation and cytokine production using enzyme-linked immunosorbent assay (ELISA). Secretion of metalloproteinases and their inhibitors was determined by zymography and ELISA, respectively. RESULTS: Monocytes bound to matrix synthesized by mesangial cells. Prestimulation of mesangial cells with tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta (TGF-beta) enhanced matrix fibronectin content (P < 0.001) and monocyte binding (P < 0.001). Blocking antibodies to fibronectin, as well as to the integrins very late antigen-4 (VLA-4) and VLA-5, reduced monocyte adhesion to mesangial matrix by approximately 50%. Incubation of monocytes with matrix, fibronectin, laminin and collagen IV enhanced production of interleukin-1beta (IL 1beta), interleukin-6 (IL-6), TNF-alpha and metalloproteinase-9 (MMP-9) when compared to cells incubated in plastic wells. However, there was no apparent difference in proliferation rate and no change in production of metalloproteinase inhibitors. CONCLUSION: Monocyte activation within the glomerulus may be mediated by binding to mesangial matrix components, particularly fibronectin. Matrix mediated activation enhances production of inflammatory cytokines and matrix degrading enzymes. PMID- 12631070 TI - IL-1beta, TNF-alpha, TGF-beta, and bFGF expression in bone biopsies before and after parathyroidectomy. AB - BACKGROUND: There is growing evidence pointing to an involvement of cytokines and growth factors in renal osteodystrophy. In this study, the expression of interleukin-l beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha), transforming growth factor-beta (TGF-beta), and basic fibroblast growth factor (bFGF) in bone biopsies taken from uremic patients before and 1 year after parathyroidectomy (PTX) was evaluated. Biochemical features and histomorphometric outcome were also studied. METHODS: Iliac bone biopsies were taken before and 1 year after PTX in nine uremic patients with severe hyperparathyroidism (HPT). Immunohistochemical techniques were used to identify the expression of IL-1 beta, TNF-alpha, TGF-beta, and bFGF in these bone samples. RESULTS: At the time of the second bone biopsy, the mean serum total alkaline phosphatase activity was normal, whereas mean serum intact parathyroid hormone (iPTH) level was slightly above the upper limit of normal values. Histomorphometric analysis showed a decrease in resorption parameters and static bone formation parameters after PTX. Dynamically, mineral apposition rate (MAR) and mineralization surface (MS/BS) decreased significantly. There was a marked local expression of IL-1beta, TNF alpha, TGF-beta, and bFGF in bone biopsies before PTX, particularly in fibrous tissue and resorption areas. One year after PTX, IL-1beta decreased from 23.6 +/- 7.5% to 9.9 +/- 3.1%, TNF-alpha from 4.5 +/- 1.5% to 0.7 +/- 0.8%, TGF-beta from 49.6 +/- 9.8% to 15.2 +/- 4.6%, and bFGF from 50.9 +/- 12.7% to 12.9 +/- 7.9% (P < 0.001). A significant correlation was documented between cytokines and growth factors expression in bone with iPTH levels before and after PTX (P < 0.05). CONCLUSIONS: Based on these results, we suggest that IL-1beta, TNF-alpha, TGF beta, and bFGF are involved in bone remodeling regulation, acting as local effectors, possibly under the control of PTH. PMID- 12631071 TI - Modulation of renal epithelial barrier function by mitogen-activated protein kinases (MAPKs): mechanism of cyclosporine A-induced increase in transepithelial resistance. AB - BACKGROUND: Cyclosporine A (CsA) has been shown to increase transepithelial resistance in Madin-Darby canine kidney (MDCK) cells, and the mechanism may involve altered phosphorylation of junctional proteins. In this study, we examine the effect of the extracellular signal-regulated protein kinase (ERK) 1/2 and p38 mitogen-activated protein kinase (MAPK) pathways on the basal transepithelial resistance (TER) and on the CsA-induced increase in TER across MDCK monolayers. Here we present evidence that CsA may be mediating some of its effects through activation of the ERK 1/2 MAPK pathway. METHODS: MDCK cells were treated with CsA (4.2 micromol/L) and paracellular permeability was assessed by measuring TER. The role of the ERK 1/2 and the p38 MAPK pathways in modulating TER was investigated using the inhibitors PD98059 and U0126 for ERK 1/2 and SB203580 for p38. ERK 1/2 and p38 phosphorylation/activation was also examined by Western blot analysis. RESULTS: CsA (4.2 micromol/L) increased the TER of MDCK monolayers. The ERK 1/2 inhibitor PD98059 decreased basal TER and also ameliorated the CsA-induced increase in TER. Similar results were found with the U0126 inhibitor of ERK 1/2. The p38 inhibitor SB203580 had no effect on the basal TER of the monolayers, however, SB203580 significantly augmented the CsA-induced increase in TER. CsA was shown to significantly activate ERK 1/2 and this activation by CsA was prevented by PD98059. Inhibition of the p38 pathway by SB203580 also resulted in activation of ERK 1/2 and this activation of ERK 1/2 was further enhanced by CsA. No effect of CsA or the inhibitors PD98059 or SB203580 on p38 phosphorylation was detected. CONCLUSION: The results presented here suggest that activation of the ERK 1/2 MAPK cascade is important in the regulation of the paracellular permeability in MDCK cells. Activation of this pathway appears to be pivotal to the CsA-induced increase in TER. PMID- 12631073 TI - Dose-dependent effects of strontium on bone of chronic renal failure rats. AB - BACKGROUND: We previously reported on increased bone strontium (Sr) levels in dialysis patients with osteomalacia versus those presenting other types of renal osteodystrophy. A causal role of strontium in the development of osteomalacia was established in a chronic renal failure (CRF) rat model. METHODS: In the present study we investigated whether the effect of Sr on bone was related to dosage. Four groups of CRF rats were studied: a control group (control-CFR; N=6) not receiving strontium and three groups of animals loaded orally with Sr during 18 weeks by adding the element as the SrCl2. H20 compound to the drinking water at concentrations of 0.03 g/100mL (Sr-30; N=6), 0.075 g/100mL (Sr-75; N=6), or 0.15 g/100mL (Sr-150; N=6) respectively. A fifth group consisting of seven animals with intact renal function (control-NRF), not receiving Sr served as controls for the effect of CRF on bone histology. RESULTS: As compared to the control-NRF and control-CRF groups, Sr administration resulted in a dose-dependent increase in bone and serum Sr levels. No difference in body weight and biochemical serum and urinary parameters [i.e., calcium (Ca), phosphorus (P), and creatinine] was noted between the various CRF groups. At sacrifice, intact parathyroid hormone (iPTH) levels of CRF groups were significantly (P < 0.05) higher than the values measured in the control-NRF group indicating the development of hyperparathyroidism secondary to the installation of the CRF. This is further supported by the differences in bone histomorphometry between the control-CRF and control-NRF animals, which, respectively, showed an increased amount of osteoid (mean +/- SEM 3.4 +/- 1.2% vs. 0.37 +/- 0.14%, P < 0.05) in combination with a distinct osteoblastic activity (35 +/- 11% vs. <2%, P < 0.05) and an increased bone formation rate [(BFR), 677 +/- 177 microm 2/mm2/day vs. 130 +/- 50 microm 2/mm2/day, P < 0.05]. Bone surface area and erodic perimeter did not differ between the various study groups. In the Sr-30 group, Sr loading went along with a dramatic reduction of the BFR as indicated by the total absence of double tetracyclin labels and osteoblastic activity, which in the presence of a low to normal amount of osteoid (2.7 +/- 1.9%) points to the development of the adynamic type of renal osteodystrophy. Interestingly, compared to the control-CRF group, histodynamic and histologic parameters of the Sr-75 group did not differ significantly and a substantial osteoblastic activity (7.6 +/- 4.0%) was seen also. In the Sr-150 group, the various osteoid parameters were significantly (P < 0.05) increased vs. all other groups and were accompanied by a reduced BFR and mineral apposition rate (MAR) and an increased mineralization lag time (MLT), indicating a mineralization defect and the development of osteomalacia. CONCLUSIONS: Our findings indicate that the role of Sr in the development of bone lesions in renal failure is complex and that, depending on the dose, the element may act via multiple pathways. PMID- 12631072 TI - Rapamycin is an effective inhibitor of human renal cancer metastasis. AB - Rapamycin is an effective inhibitor of human renal cancer metastasis. BACKGROUND: Human renal cell cancer (RCC) is common and is 10 to 100 times more frequent in patients with end-stage renal disease (ESRD) and candidates for renal transplantation. Treatment of metastatic RCC is largely ineffective and is further undermined by immunosuppressive therapy in transplant recipients. A treatment regimen that prevents transplant rejection while constraining RCC progression would be of high value. METHODS: We developed a human RCC pulmonary metastasis model using human RCC 786-O as the tumor challenge and the severe combined immunodeficient (SCID) beige mouse as the host. We explored the effect of rapamycin, cyclosporine, or rapamycin plus cyclosporine on the development of pulmonary metastases and survival. The effects of the drugs on tumor cell growth, apoptosis, and expression of vascular endothelial growth factor (VEGF-A) and transforming growth factor beta1 (TGF-beta1) were also investigated. RESULTS: Rapamycin reduced, whereas cyclosporine increased, the number of pulmonary metastases. Rapamycin was effective in cyclosporine-treated mice, and rapamycin or rapamycin plus cyclosporine prolonged survival. Rapamycin growth arrested RCC 786-O at the G1 phase and reduced VEGF-A expression. Immunostaining of lung tissues for von Willebrand factor was minimal and circulating levels of VEGF-A and TGF-beta1 were lower in the rapamycin-treated mice compared to untreated or cyclosporine-treated mice. CONCLUSION: Our findings support the idea that rapamycin may be of value for patients with RCC and that its antitumor efficacy is realized by cell cycle arrest and targeted reduction of VEGF-A and TGF-beta1. A regimen of rapamycin and cyclosporine, demonstrated to be effective in reducing acute rejection of renal allografts, may prevent RCC progression as well, and has the potential to prevent mortality due to RCC in patients with ESRD who have received renal allografts. PMID- 12631074 TI - Shedding of growth-suppressive gangliosides from glomerular mesangial cells undergoing apoptosis. AB - BACKGROUND: Apoptosis of glomerular mesangial cells is a common feature in several types of glomerular diseases. However, its pathophysiologic significance is not known. We recently identified gangliosides as a major growth-inhibitory substance in the conditioned medium of mesangial cells. In this report, we tested whether biologically distinct forms of cell fate, apoptosis and necrosis, could modulate ganglioside shedding from mesangial cells. METHODS: Mesangial cells were exposed to low (10 to 40 mJ/cm2) and high (400 mJ/cm2) doses of ultraviolet light to induce apoptosis and necrosis, respectively. Conditioned media were collected and examined for its growth-inhibitory activity for mesangial cells. Ganglioside shedding was analyzed using metabolic labeling and thin-layer chromatography (TLC). RESULTS: Shedding of gangliosides as well as growth-inhibitory activity in the conditioned medium predominantly increased when mesangial cells were undergoing apoptosis in contrast to that of viable or necrotic mesangial cells. The inhibitory substance in the conditioned medium from apoptotic mesangial cells completely fulfilled the characteristic criteria of gangliosides. This substance was less than 3 kD and was sensitive to neuraminidase digestion. Shedding of gangliosides from mesangial cells reduced significantly when apoptosis was inhibited by overexpression of antiapoptotic gene, Bcl-XL. In addition, ganglioside shedding also increased when mesangial cells were exposed to other inducers of apoptosis for mesangial cells (i.e., H2O2 and staurosporin). CONCLUSION: These results provide the novel link between masangial cell apoptosis and increased release of gangliosides that potentially suppress mesangial cell proliferation and thus indicate a mechanism for the negative regulation of mesangial cell growth by apoptosis. PMID- 12631075 TI - Methylglyoxal induces apoptosis through activation of p38 mitogen-activated protein kinase in rat mesangial cells. AB - BACKGROUND: The formation of methylglyoxal (MG), a highly reactive dicarbonyl compound, is accelerated through several pathways, including the glycation reaction under diabetic conditions, presumably contributing to tissue injury in diabetes. On the other hand, apoptotic cell death of glomerular cells has been suggested to play a role in the development of glomerulosclerosis in various types of glomerular injuries. We therefore examined whether MG was capable of inducing apoptosis in rat mesangial cells to address the possible mechanism by which hyperglycemia-related products accelerated pathologic changes in diabetic glomerulosclerosis. METHODS: Rat mesangial cells were incubated with 0 to 400 micromol/L MG, followed by the detection of apoptosis by both TUNEL method and electrophoretic analysis for DNA fragmentation. In addition, we investigated intracellular mechanisms mediating MG-induced apoptosis, focusing especially on the p38 mitogen-activated protein kinase (MAPK) pathway. RESULTS: MG induced apoptosis in rat mesangial cells in a dose-dependent manner and was accompanied by the activation of p38alpha isoform. Aminoguanidine and N-acetyl-l-cysteine inhibited the MG-induced p38 MAPK activation, as well as apoptosis in rat mesangial cells, suggesting the involvement of oxidative stress in these phenomena. SB203580, a specific inhibitor of p38 MAPK also suppressed the MG induced apoptosis in rat mesangial cells. CONCLUSIONS: These results suggest a potential role for MG in glomerular injury through p38 MAPK activation under diabetic conditions and may serve as a novel insight into the therapeutic strategies for diabetic nephropathy. PMID- 12631076 TI - High glucose activates the p38 MAPK pathway in cultured human peritoneal mesothelial cells. AB - BACKGROUND: Peritoneal fibrosis is a serious complication in long-term continuous ambulatory peritoneal dialysis (CAPD) patients, but the underlying mechanism is not well understood. Since high glucose activates the p38 mitogen-activated protein kinase (MAPK) pathway in various kinds of cells, and because mesothelial cells are always exposed to high glucose dialysate, we examined the activity and expression of p38 MAPK members in cultured human peritoneal mesothelial cells (HPMCs) under high glucose conditions. METHODS: HPMCs were isolated from omentum and subcultured. After serum restriction, HPMCs were exposed to 5.6 mmol/L glucose (low glucose), 5.6 mmol/L glucose + 34.5 mmol/L mannitol (low glucose + mannitol), or 40 mmol/L glucose (high glucose) for 3 minutes to 48 hours with or without SB203580. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were performed to determine mRNA and protein expression, respectively. RESULTS: p38 MAPK and cyclic adenosine monophosphate (cAMP) responsive element binding protein (CREB) activities and mRNA expressions were significantly increased in HPMCs exposed to high glucose compared to low glucose or low glucose + mannitol after 10 minutes and remain at higher levels to 48 hours (P < 0.05), but total p38 MAPK and CREB protein expressions did not differ. MAPK kinase 3/6 (MKK3/6) activity and mRNA expression were also higher in high glucose cells after 3 minutes (P < 0.05), and fibronectin mRNA expression was significantly increased in HPMCs exposed to high glucose after 2 hours (P < 0.05). In contrast, high glucose significantly inhibited MAPK phosphatase-1 (MKP 1) protein and mRNA expression after 10 minutes (P < 0.05). SB203580 (1 micromol/L) pretreatment for 1 hour significantly reduced high glucose-induced CREB activity and fibronectin mRNA expression by 89% and 75%, respectively (P < 0.05). CONCLUSION: p38 MAPK activity was increased in HPMCs exposed to high glucose, in parallel with increased MKK3/6 activity and decreased MKP-1 expression, resulting in CREB activation. This activated p38 MAPK pathway may play a role in the pathogenesis of peritoneal fibrosis. PMID- 12631077 TI - Expression of the K+ channel Kir7.1 in the developing rat kidney: role in K+ excretion. AB - BACKGROUND: Coordinated expression of ROMK (luminal K+ channel in the thick ascending limb and the collecting duct) and Na+,K+-ATPase has been demonstrated to be involved in the postnatal development of renal K+ excretion; however, the developmental expression of the basolateral K+ channel Kir7.1 is unknown. The purpose of this study was to elucidate the possible involvement of Kir7.1 in the maturation of renal K+ excretion. METHODS: Developmental changes in the renal K+ excretion under the condition of K+ overload was investigated by collecting urine from neonatal rats infused with K+ (KCl solution). RNase protection analysis was used to elucidate the expression of Kir7.1 and Na+,K+-ATPase mRNA in pre- and postnatal rats, and the expression of Kir7.1 and ROMK mRNA at 7, 14, and 21 days. Western blotting of Kir7.1, and immunohistochemistry of Kir7.1 and ROMK were used to determine their protein expression. RESULTS: The ratio of urinary K+ excretion to K+ load increased between 7 and 14 days after birth. In addition, half excretion time of K+ load gradually decreased through the experimental period of 7 and 21 days. Na+,K+-ATPase mRNA levels showed a peak of up-regulation at birth that remained elevated. ROMK1 mRNA levels significantly increased between 7 and 14 days. On the other hand, Kir7.1 mRNA and protein levels significantly increased between 14 and 21 days. Kir7.1 protein in the thick ascending limb was first recognized at 7 days, whereas its expression in the distal convoluted tubule and the cortical collecting duct was found in 21-day-old neonates. CONCLUSION: Our results suggest that Kir7.1 is involved in the development of renal K+ excretion between 14 and 21 days after birth under the condition of K+ overload. PMID- 12631078 TI - Sulfate conjugating and transport functions of MDCK distal tubular cells. AB - BACKGROUND: Transfected Madin-Darby canine kidney (MDCK) cells (of distal tubular origin) have been used to study transport of organic anions. These cells have not been shown to possess sulfate-conjugating activity. Neither has transport activity been demonstrated in nontransfected MDCK cells. METHODS: Polarized and monolayers of nontransfected MDCK type II cells were incubated with prototype substrates of phenolsulfotransferase (PST) and sodium sulfate in the absence or presence of known inhibitors of multidrug resistance protein (MRP): (3-3-(2-(7 chloro-2-quinionlinyl) ethenyl)phenyl)(3-dimethylamino-3 oxopropyl)thio)methyl)thio) propanoic acid (MK571), cyclosporin A (CsA), and probenecid. Effects of glutathione (GSH) and buthionine sulfoximine (BSO), potential modulators of the organic anion transporting protein/polypeptide (OATP) isoform, OATP1 were also examined. Sulfated conjugates were identified by high performance liquid chromatography (HPLC)-radiometry or HPLC-fluorimetry. RESULTS: Uptake, sulfate conjugation, and efflux of the sulfated conjugates of harmol, p nitrophenol, N-acetyldopamine and acetaminophen were demonstrated. Activities in MDCK type II cells were higher than those in HepG2, human fetal liver, and Chang liver cells. A significant decrease in extracellular with a reciprocal increase in intracellular harmol sulfate was observed with MK571, CsA, and probenecid and with preloading of glutathione. Depletion of intracellular glutathione by BSO had the opposite effects. CONCLUSIONS: Normal (nontransfected) MDCK type II cells provide a suitable system for the study of the physiologic processes of uptake, sulfate conjugation, and transport of sulfated conjugates in kidney cells. Based on the action of specific inhibitors and modulators of MRP2 and OATP1, it was concluded that MRP2-like and OATP1-like transporters are possibly responsible for the transport of sulfated conjugates. PMID- 12631079 TI - Glomerular autacoids stimulated by bradykinin regulate efferent arteriole tone. AB - BACKGROUND: We have shown that when efferent arterioles are perfused retrograde to avoid the influence of vasoactive autacoids released by the glomerulus, bradykinin causes dilatation via release of cytochrome p450 (cp450) metabolites, probably epoxyeicosatrienoic acids (EETs). Here we tested the hypothesis that the glomerulus releases cyclooxygenase (COX) and cp450 metabolites. These eicosanoids, acting as vasopressor and vasodepressor autacoids, control efferent arteriole resistance downstream from the glomerulus. METHODS: Rabbit efferent arterioles were perfused orthograde through the glomerulus from the end of the afferent arteriole to determine whether bradykinin induces the release of glomerular autacoids that influence efferent arteriole resistance. Efferent arterioles were preconstricted with norepinephrine, and increasing doses of bradykinin were added to the perfusate in the presence or absence of COX and cp450 inhibitors. RESULTS: When efferent arterioles were perfused orthograde through the glomerulus, bradykinin at 10 nmol/L caused significant and reproducible dilatation; diameter increased from 8.0 +/- 0.5 to 12.6 +/- 0.4 microm (P < 0.05). This effect was not modified by a nitric oxide synthase (NOS) inhibitor. In the presence of indomethacin, a COX inhibitor, bradykinin-induced dilatation was almost completely blocked (from 8.0 +/- 0.5 to 9.3 +/- 0.6 microm). This blockade was completely reversed by 20-hydroxyeicosa-6(Z),15(Z) dienoic acid (20-HEDE), a specific antagonist of the vasoconstrictor cp450 metabolite 20-hydroxyeicosatetraenoic acid (20-HETE); diameter increased from 6.6 +/- 0.7 to 13.2 +/- 0.5 microm. To test the hypothesis that this dilatation was due to EETs, a specific inhibitor of EET synthesis, N-methylsulphonyl-6-(2 proparglyloxyphenyl)hexanamide (MS-PPOH), was added to the arteriolar perfusate. In the presence of indomethacin and 20-HEDE, bradykinin caused dilatation and this effect was completely blocked by MS-PPOH (1 microm) (from 7.6 +/- 0.6 to 7.3 +/- 0.5 microm). CONCLUSIONS: We concluded that in response to bradykinin, the glomerulus releases COX metabolites (probably prostaglandins) that have a vasodilator effect. When COXs are inhibited, the vasoconstrictor 20-HETE released by the glomerulus is able to oppose the vasodilator effect of bradykinin. This vasodilator effect is mediated by EETs released by the glomerulus and/or the efferent arteriole and does not involve nitric oxide. The balance between these opposing effects of various eicosanoids controls efferent arteriole resistance downstream from the glomerulus. PMID- 12631080 TI - Mycophenolate mofetil prevents arteriolopathy and renal injury in subtotal ablation despite persistent hypertension. AB - BACKGROUND: Although renal protective effect of interrupting the inflammatory process is well established, it is still controversial if it also prevents the glomerular hemodynamic disturbances that initiate renal injury. We investigated the effects of suppressing inflammation with mycophenolate mofetil (MMF) on glomerular hemodynamics, arteriolar structural changes, and renal histologic injury in rats with subtotal renal ablation METHODS: Micropuncture studies were performed 30 days after 5/6 nephrectomy in rats untreated and treated with MMF (30 mg/kg/day). Renal histology, immunohistochemistry for lymphocytes, macrophages and inducible nitric oxide synthase (iNOS) expression, as well as afferent arteriolar (AA) morphometry was evaluated. RESULTS: Renal ablation significantly increased proteinuria (6.8 to 82.7 mg/day), mean arterial pressure (MAP) (120 to 166 mm Hg), single-nephron glomerular filtration rate (SNGFR) (34.8 to 56.3 nL/min), glomerular plasma flow (QA) (117.7 to 246.9 nL/min), and glomerular capillary pressure (PGC) (48.9 to 61.0 mm Hg). Afferent resistance (AR), efferent resistance, and ultrafiltration coefficient remained unchanged. Despite persisting arterial hypertension (152 mm Hg), MMF prevented proteinuria (13.3 mg/day), and significantly reduced SNGFR (44.4 nL/min), PGC (49.1 mm Hg), and QA (163.2 nL/min) due to a rise in AR (3.13 vs. 2.18 1010 dyn/sec/cm-5). Glomerular sclerosis, tubulointerstitial damage, lymphocyte and macrophage infiltration, and iNOS expression were significantly reduced by MMF, in addition hypertrophy of AA resistance evaluated by the media/lumen ratio was prevented (P < 0.001). CONCLUSIONS: Reduction in proteinuria, SNGFR, QA, and PGC, despite elevated MAP, indicate preservation of AA function. These results suggest that inflammation associated arteriolopathy of AA contributes to glomerular hemodynamic disturbances that participate in the progression of renal disease. PMID- 12631081 TI - Uremia induces the osteoblast differentiation factor Cbfa1 in human blood vessels. AB - BACKGROUND: Bone matrix proteins are expressed in calcified arteries from dialysis patients, suggesting that vascular smooth muscle cells (VSMCs) may transform to osteoblast-like cells. One of the key transcriptional regulators of osteoblast differentiation is Cbfa1. Thus, we hypothesized that this may be a key factor in arterial calcification. METHODS: To test this hypothesis, we examined sections of the inferior epigastric artery from uremic patients for the presence of Cbfa1 and type I collagen and osteopontin by in situ hybridization and immunostaining. We also examined the effect of pooled uremic sera from dialysis patients on the expression of Cbfa1 by reverse transcription-polymerase chain reaction (RT-PCR) in bovine VSMCs in vitro. RESULTS: Cbfa1 and osteopontin were expressed in both the media and the intima in vessels that were calcified, but there was only minimal staining in non-calcified vessels. In vitro studies demonstrated that pooled uremic serum, compared to pooled control human serum induced the expression of Cbfa1 by RT-PCR in bovine VSMCs in a time-dependent, nonphosphorus-mediated mechanism. CONCLUSION: These results support that Cbfa1 is a key regulatory factor in the vascular calcification observed in dialysis patients and is up-regulated in response to many uremic toxins. PMID- 12631082 TI - Homocysteine enhances TIMP-1 expression and cell proliferation associated with NADH oxidase in rat mesangial cells. AB - BACKGROUND: Recent studies in our laboratory demonstrated that chronic hyperhomocysteinemia (hHcys) induced glomerular sclerosis. The mechanism mediating hHcys-induced glomerular damage remains unknown. The present study was designed to test a hypothesis that homocysteine (Hcys) increases the production by nicotinamide adenine dinucleotide (NADH) oxidase and thereby stimulates the formation of tissue inhibitor of metalloproteinase (TIMP-1) in rat mesangial cells, consequently leads to glomerulosclerosis. METHODS: Rat mesangial cells were incubated with L-homocysteine (L-Hcys) to determine the effects of Hcys on cell proliferation and metabolism of extracellular matrix (ECM). Northern blot, Western blot, oligonucleotide transfection, measurements of NADH oxidase activity and levels, and cell proliferation assay were performed. RESULTS: In cultured rat mesangial cells, treatment with L-Hcys (40 to 160 micromol/L) markedly increased the mRNA levels of TIMP-1 and Gp91 and led to accumulation of collagen I, which were accompanied by enhanced cell proliferation and NADH oxidase activity in mesangial cells. These Hcys-induced biochemical and functional changes were substantially blocked by a NADH oxidase inhibitor, diphenylene iodonium chloride (DPI) or a superoxide dismutase (SOD) mimetic, hydroxyl-tetramethylpiperidin-oxyl (TEMPOL). Moreover, blockade of NADH oxidase subunit, phox22, by its antisense oligodeoxynucleotide also eliminated the increase in NADH oxidase activity induced by L-Hcys. CONCLUSION: These results indicate that Hcys-induced alterations of ECM metabolism in mesangial cells are associated with enhanced NADH oxidase activity and that oxidative stress-stimulated up-regulation of TIMP 1 may play an important role in the deposition of collagen or ECM elements in the glomeruli during hHcys. PMID- 12631084 TI - Morphometric study of arterioles and glomeruli in the aging kidney suggests focal loss of autoregulation. AB - BACKGROUND: In the past it was widely assumed that hyaline afferent arteriolosclerosis was responsible for ischemic glomerulosclerosis in the aging and hypertensive kidney. However, glomerular lesions of focal segmental glomerulosclerosis are now recognized in essential hypertension. Experimentally, such lesions are associated with loss of autoregulation of blood flow and glomerular hyperperfusion, as well as initial glomerular hypertrophy. These observations challenge the notion of ischemia as a unitary explanation for glomerulosclerosis. METHODS: A morphometric study was performed on normal portions of eight kidneys removed for tumors in aging, normotensive patients. Measurements were made of 126 pairs of afferent arterioles and their associated glomeruli. In addition, the amount of extracellular matrix (ECM) in the immediate periglomerular region was quantitated. RESULTS: Afferent arterioles were divided into three types according to the presence or absence of hyaline deposits and whether these did or did not obstruct the lumen. Arterioles with nonobstructive hyaline deposits had lumens over twice as large as those without deposits (482 +/ 240 micro2 vs. 204 +/- 160 micro2, P=0.0000). Their associated glomeruli had significantly greater total capillary area, particularly the hilar capillaries (1276 +/- 797 micro2 vs. 667 +/- 492 micro2, P=0.002), but with larger individual capillaries elsewhere as well (P=0.03). Arterioles with obstructive deposits differed from those with nonobstructive deposits by their smaller lumens (P=0.001) and walls (P=0.004), with a higher proportion of ECM in the periglomerular region (P=0.001), all consistent with a later stage of lesion. Glomeruli were divided into four basic types: normal, hypertrophic, glomeruli with features of focal segmental glomerulosclerosis (FSGS-type), and ischemic. Compared to normal glomeruli, hypertrophic glomeruli were larger, with greater total capillary area (P=0.0005), particularly the hilar capillaries (P=0.0000), and larger capillaries in the remainder of the tuft (P=0.003), but showed no evident lesions. FSGS-type glomeruli were also larger, with larger hilar capillaries (P=0.0005), but showed an increase in ECM due to sclerotic lesions (P=0.004). The remaining capillaries showed an inverse relation with the amount of mesangial matrix, showing a spectrum of sizes from enlarged to shrunken. As anticipated, ischemic glomeruli were significantly smaller than normal ones in every parameter measured. There was a strong association between hypertrophic/FSGS-type glomeruli and hyaline arteriolosclerosis, found in 90.3% of such glomeruli, versus 29.1% for the remaining glomeruli (P=0.0001). The great majority of hyaline deposits were of the nonobstructive variety (86.2%), but some were obstructive (13.8%), particularly in FSGS-type glomeruli, consistent with a more advanced lesion. CONCLUSIONS: We believe we have demonstrated in the aging kidney of humans the morphologic correlates of loss of autoregulation, occurring on a focal basis, with afferent arteriolar dilatation and increase in glomerular capillary size and subsequent focal segmental glomerulosclerosis. Hyaline arteriolosclerosis of the nonobstructive sort is strongly associated with these changes and may play a role in their pathogenesis. PMID- 12631083 TI - Sexual dimorphism in the aging kidney: Effects on injury and nitric oxide system. AB - BACKGROUND: Male gender confers enhanced susceptibility to development of age dependent kidney damage. In other models of progressive renal disease, development of injury is linked to declines in renal nitric oxide synthase (NOS) capacity. METHODS: We investigated the in vitro characteristics of the renal NOS system in young (3 to 5 months), middle-aged (11 to 13 months) and old (18 to 22 months) male and female Sprague-Dawley rats. RESULTS: NOS activity (pmol [3H] arginine converted to [3H]-citrulline/mg protein/minute) is reduced in the soluble fraction of renal cortex from old versus young males but not females. In contrast, NOS activity in the soluble fraction of cerebellum is not altered by age or gender. The abundance of endothelial NOS (eNOS) and neuronal (nNOS) is reduced in renal cortex of old versus young males but is unchanged in female cortex. In renal medulla, eNOS protein is reduced with age in both males and females. We found no difference in abundance of either eNOS or nNOS protein in the cortex of young male and female rats. The incidence and severity of glomerular damage increases markedly with age in the male and only slightly in the female. CONCLUSION: These findings indicate that a relative reduction occurs in renal NOS in the male kidney with advancing age, whereas NOS protein and activity is maintained during aging in females. This, together with the marked age-dependent kidney damage seen in the male, suggests that the renal NO deficiency in the aging male rat may contribute to the age-dependent kidney damage. PMID- 12631086 TI - Blood lead and chronic kidney disease in the general United States population: results from NHANES III. AB - BACKGROUND: High lead exposure is associated with hypertension and renal dysfunction but the effect of low-level environmental exposure is not as well studied. METHODS: We examined the association between blood lead and renal function among a representative sample of the civilian noninstitutionalized United States population with and without hypertension, age 20 years old or older, participating in the Third National Health and Nutrition Examination Survey (NHANES III) (N=15211). Elevated serum creatinine was defined as >or=99th percentile of each race-sex specific distribution for healthy young adults and chronic kidney disease (CKD) as a glomerular filtration rate (GFR) <60 mL/min estimated using the Modification of Diet in Renal Disease (MDRD) formula. RESULTS: Among persons with and without hypertension, mean blood lead was 4.21 and 3.30 ug/dL, respectively, the prevalence of elevated serum creatinine was 11.5% and 1.8%, respectively, and CKD was 10.0% and 1.1%, respectively. Among persons with hypertension, a graded association was present between higher quartile of blood lead and a higher odds ratio of both an elevated serum creatinine and CKD. Comparing the highest to lowest quartile of blood lead, the multivariate adjusted odds ratio (95% CI) of an elevated serum creatinine and CKD were 2.41 (1.46, 3.97) and 2.60 (1.52, 4.45), respectively. The analogous adjusted odds ratios (95% CI) among normotensives were 1.09 (0.53, 2.22) and 1.09 (0.41, 2.89), respectively. Associations were consistent when modeling lead as a continuous variable and in all subgroups except smokers. CONCLUSION: In the United States population with hypertension, exposure to lead, even at low levels, is associated with CKD. Reduction of lead exposure may reduce the burden of CKD in the community. PMID- 12631085 TI - Dietary risk factors for hyperoxaluria in calcium oxalate stone formers. AB - BACKGROUND: Hyperoxaluria is a major predisposing factor in calcium oxalate urolithiasis. The aim of the present study was to clarify the role of dietary oxalate in urinary oxalate excretion and to assess dietary risk factors for hyperoxaluria in calcium oxalate stone patients. METHODS: Dietary intakes of 186 calcium oxalate stone formers, 93 with hyperoxaluria (>or=0.5 mmol/day) and 93 with normal oxalate excretion (<0.4 mmol/day), were assessed by a 24-hour weighed dietary record. Each subject collected 24-hour urine during the completion of the food record. Oxalate content of foods was measured by a recently developed analytical method. RESULTS: The mean daily intakes of energy, total protein, fat and carbohydrates were similar in both groups. The diets of the patients with hyperoxaluria were estimated to contain 130 mg/day oxalate and 812 mg/day calcium as compared to 101 mg/day oxalate and 845 mg/day calcium among patients without hyperoxaluria. These differences were not significant. The mean daily intakes of water (in food and beverages), magnesium, potassium, dietary fiber and ascorbic acid were greater in patients with hyperoxaluria than in stone formers with normal oxalate excretion. Multiple logistic regression analysis revealed that urinary oxalate excretion was significantly associated with dietary ascorbate and fluid intake, and inversely related to calcium intake. Differences of estimated diet composition of both groups corresponded to differences in urinary parameters. CONCLUSIONS: These findings suggest that hyperoxaluria predominantly results from increased endogenous production and from intestinal hyperabsorption of oxalate, partly caused by an insufficient supply or low availability of calcium for complexation with oxalate in the intestinal lumen. PMID- 12631087 TI - High-dose intravenous melphalan with autologous stem cell transplantation in AL amyloidosis-associated end-stage renal disease. AB - BACKGROUND: The development of end-stage renal disease (ESRD) is common among patients with amyloid light-chain AL amyloidosis-associated renal disease and survival of these patients is poor. High-dose intravenous melphalan and autologous stem cell transplantation induce remission of the plasma cell dyscrasia in a significant proportion of patients with AL amyloidosis. The efficacy and tolerability of such treatment for patients with AL amyloidosis associated ESRD are unknown. METHODS: Between June 1994 and June 2000, 15 patients with AL amyloidosis-associated ESRD were treated with intravenous melphalan (70 to 200 mg/m2) and autologous peripheral blood stem cell transplantation. Clinical and laboratory data were prospectively collected prior to treatment, during the peritransplant period, and at 3 months, 12 months, and annually thereafter. Treatment outcomes and toxicities were compared with 180 non ESRD patients treated during the study period. RESULTS: Eight of 15 patients (53%) had a hematologic complete response following treatment. Two patients (13%) died during the peritransplant period. Transfusion requirements were greater and there was a trend toward increased severity of mucositis in the ESRD patients compared with the non-ESRD patients. Median survival for the ESRD patients with a hematologic complete response was 4.5 years. Five patients with hematologic complete response have either undergone or are awaiting renal transplantation. CONCLUSION: High-dose intravenous melphalan with stem cell transplantation is an effective treatment in selected patients with AL amyloidosis-associated ESRD. Although the toxicity profile is greater in ESRD patients, the treatment offers the possibility of successful renal transplantation if hematologic remission is achieved. This treatment should be considered for patients with AL amyloidosis associated ESRD. PMID- 12631088 TI - Cardiac disease in young adult patients with end-stage renal disease since childhood: a Dutch cohort study. AB - BACKGROUND: Cardiovascular disease is the most important cause of death in patients with pediatric end-stage renal disease (ESRD). Yet, few data exist on cardiac function in these patients. We assessed the extent of cardiac abnormality and analyzed its association with potential determinants in young adult patients with pediatric ESRD in a long-term follow-up study. METHODS: All Dutch living adult patients with ESRD onset at age of 0 to 14 years between 1972 and 1992 were invited for echocardiography and blood pressure assessment. Special attention was paid to evidence of left ventricular hypertrophy (LVH), diastolic dysfunction, and aortic valve calcification. We collected data on determinants by review of all medical charts. RESULTS: Of all the 187 living patients, 140 participated in the study. Of those, 110 patients had received a transplant and 30 patients were on dialysis. Mean age was 29.2 (20.7 to 41.8) years. Left ventricular mass index (LVMI) exceeded 150 g/m2 in 47% of all male patients and 120 g/m2 in 39% of all female patients, both consistent with LVH. Diastolic dysfunction, defined as an early over atrial transmitral blood flow velocity (E/A ratio) <1, was found in 18 (13%) patients; 27 (19%) had aortic valve calcification. Multiple regression analysis revealed the following: a high LVMI was associated with a current high blood pressure (beta=0.46, P < 0.001) and male gender (beta=0.21, P=0.009), a low E/A ratio with aging (beta=-0.28, P < 0.001) and a glomerular filtration rate (GFR) <25 mL/min/1.73 m2 (beta=-0.29, P < 0.001), and aortic valve calcification with prolonged peritoneal dialysis (beta=0.36, P < 0.001). CONCLUSION: Young adult patients with pediatric ESRD are at risk for LVH caused by hypertension and for aortic valve calcification. Diastolic function decreases with age and is enhanced by a current low GFR. Prolonged peritoneal dialysis may enhance aortic valve calcification. PMID- 12631089 TI - Effect of vitamin C supplements on urinary oxalate and pH in calcium stone forming patients. AB - BACKGROUND: The contribution of ascorbate to urinary oxalate is controversial. The present study aimed to determine whether urinary oxalate and pH may be affected by vitamin C supplementation in calcium stone-forming patients. METHODS: Forty-seven adult calcium stone-forming patients received either 1 g (N=23) or 2 g (N=24) of vitamin C supplement for 3 days and 20 healthy subjects received 1 g. A 24-hour urine sample was obtained both before and after vitamin C for calcium, oxalate, magnesium, citrate, sodium, potassium, and creatinine determination. The Tiselius index was used as a calcium oxalate crystallization index. A spot fasting morning urine sample was also obtained to determine the urinary pH before and after vitamin C. RESULTS: Fasting urinary pH did not change after 1 g (5.8 +/ 0.6 vs. 5.8 +/- 0.7) or 2 g vitamin C (5.8 +/- 0.8 vs. 5.8 +/- 0.7). A significant increase in mean urinary oxalate was observed in calcium stone forming patients receiving either 1 g (50 +/- 16 vs. 31 +/- 12 mg/24 hours) or 2 g (48 +/- 21 vs. 34 +/- 12 mg/24 hours) of vitamin C and in healthy subjects (25 +/- 12 vs. 39 +/- 13 mg/24 hours). A significant increase in mean Tiselius index was observed in calcium stone-forming patients after 1 g (1.43 +/- 0.70 vs. 0.92 +/- 0.65) or 2 g vitamin C (1.61 +/- 1.05 vs. 0.99 +/- 0.55) and in healthy subjects (1.50 +/- 0.69 vs. 0.91 +/- 0.46). Ancillary analyses of spot urine obtained after vitamin C were performed in 15 control subjects in vessels with or without ethylenediaminetetraacetic acid (EDTA) with no difference in urinary oxalate between them (28 +/- 23 vs. 26 +/- 21 mg/L), suggesting that the in vitro conversion of ascorbate to oxalate did not occur. CONCLUSION: These data suggest that vitamin C supplementation may increase urinary oxalate excretion and the risk of calcium oxalate crystallization in calcium stone-forming patients. PMID- 12631090 TI - Gentamicin pharmacokinetics during slow daily home hemodialysis. AB - BACKGROUND: Gentamicin is commonly used in hemodialysis patients. Gentamicin pharmacokinetics during traditional hemodialysis have been described. Slow daily home (SDH) hemodialysis (7 to 9 hours a day/6 days a week) use is increasing due to benefits observed with increased hemodialysis. We determined gentamicin pharmacokinetics for SDH hemodialysis patients. METHODS: Eight patients (four male and four female) received a single intravenous dose of 0.6 mg/kg gentamicin post-hemodialysis. Blood samples were collected at 5, 10, 15, 30, and 60 minutes after dose. The next day patients underwent a typical SDH hemodialysis (high-flux F50NR dialyzer) session. Blood samples were taken at 0, 5, 15, 60, 120, 240, 360, 480 minutes during and 15, 30 and 60 minutes post-hemodialysis. Baseline and 24 hour urine samples were collected. Pharmacokinetic parameters were calculated assuming a one-compartment model. RESULTS: Patients were 42.5 +/- 13.1 years old (mean +/- SD). Inter-, intra-, and post-hemodialysis collection periods were 17.0 +/- 2.1 hours, 8.1 +/- 0.4 hours, and 1.1 +/- 0.1 hours, respectively. Intra-, and interdialytic gentamicin half-lives were different (intradialytic, 3.7 +/- 0.8 hours; interdialytic, 20.4 +/- 4.7 hours; P < 0.0001). Hemodialysis clearance accounted for 70.5% gentamicin total clearance. Renal clearance correlated with glomerular filtration rate (GFR) (renal clearance=1.2 GFR; r2=0.98; P < 0.001). Mean peak and trough of hemodialysis concentrations were 1.8 +/- 0.6 microg/mL and 0.5 +/- 0.2 microg/mL, respectively. Post-hemodialysis rebound was 3.1 +/- 8.8% at 1 hour. CONCLUSION: Pharmacokinetic model predicts 2.0 to 2.5 mg/kg dose gentamicin post-hemodialysis would provide peak (1 hour post-dose) and trough (end of SDH hemodialysis session) concentrations of 6.0 to 7.5 microg/mL and 0.7 to 0.8 microg/mL, respectively. This would provide adequate coverage for most gram-negative organisms in SDH hemodialysis patients. PMID- 12631091 TI - Serial ANCA titers: useful tool for prevention of relapses in ANCA-associated vasculitis. AB - BACKGROUND: The value of measuring serial antineutrophil cytoplasmic autoantibody (ANCA) titers in guiding therapy among patients with ANCA-associated vasculitis is controversial. METHODS: We measured serial titers of proteinase 3 (PR3)- and myeloperoxidase (MPO)-ANCA by antigen-specific enzyme-linked immunosorbent assays (ELISAs) in 48 patients with ANCA-associated vasculitis who were followed up during remission at the Massachusetts General Hospital from 1990 through 2000 (mean follow-up, 46.2 months). We retrospectively assessed disease activity by Birmingham Vasculitis Activity Score (BVAS). RESULTS: We found 21 episodes of fourfold or greater ANCA titer rises in 17 patients who were in complete remission (BVAS=0). Among eight patients who had 10 such titer rises and were not given increased immunosuppression, (group I), all suffered relapses after each episode (mean interval, 5.8 months), whereas among 11 patients, each with one titer rise, who received preemptive increased immunosuppression, (group II), only two relapses occurred, at 3 and 6 months. The difference in the cumulative incidence of relapses in a 1-year period between the two groups was 82% (P=0.0002). Changes in ANCA titers were also used to help guide therapy in the other 31 patients in the study; patients with slight titer rises often received incremental increases in immunosuppression, whereas those with falling titers received incremental decreases. The overall outcome in the entire group was favorable; 46 patients were alive at the end of the study; two died of unrelated diseases. CONCLUSION: Serial measurements of PR3- and MPO-ANCA titers in patients with ANCA-associated vasculitis during remission can help predict relapses, and preemptive increases in immunosuppression following fourfold titer rises reduces the risk of relapses. Moreover, adjustment of immunosuppression based on lesser titer changes appears to result in a favorable outcome. PMID- 12631092 TI - Transferrin saturation versus reticulocyte hemoglobin content for iron deficiency in Japanese hemodialysis patients. AB - BACKGROUND: Iron deficiency is a frequent cause of recombinant human erythropoietin (rhEPO)-resistant anemia in hemodialysis patients. Both reticulocyte hemoglobin content (CHr) and transferrin saturation (TSAT) have been proposed as markers of iron deficiency, but it is unclear which parameter is superior. METHODS: To compare the efficacy of CHr and TSAT as an indicator for treatment of iron deficiency, we conducted a single-center, open-label, prospective, randomized, controlled trial at the Kidney Center in Shinraku-en Hospital of 197 Japanese patients on chronic hemodialysis. After 4 weeks of run in period during which iron supplementation was suspended, 100 patients who were randomized to the CHr group received 240 mg iron colloid intravenously over 2 weeks when CHr less than 32.5 pg, and 97 patients who were randomized to the TSAT group received the same doses of iron colloid when TSAT less than 20%. We measured the rhEPO dose needed to maintain prestudy hematocrit levels, hematocrit, CHr, TSAT, serum ferritin, percentage of hypochromic red blood cells, and total iron administered. RESULTS: Sixteen weeks later, 94 patients in the CHr group and 89 patients in the TSAT group finished the study. The doses of rhEPO required decreased by 35.8% (4081 to 2629 U/week, P < 0.005) in the TSAT group, but not significantly in the CHr group (4121 to 3606 U/week). Although CHr increased promptly after the iron administration in both groups, TSAT increased only in the TSAT group. CONCLUSIONS: Although CHr reflects the iron status more sensitively, TSAT is a better clinical marker for iron supplementation therapy. PMID- 12631093 TI - Effects of combined ACE inhibitor and angiotensin II antagonist treatment in human chronic nephropathies. AB - BACKGROUND: Proteinuria predicts renal disease progression, and its reduction by angiotensin-converting enzyme inhibitors (ACEi) or angiotensin II receptor antagonists (ARA) is renoprotective. METHODS: In this prospective, randomized, cross-over study of 24 patients with nondiabetic, chronic nephropathies, we compared the effects on proteinuria, renal hemodynamics, and glomerular permselectivity of 8 weeks with comparable blood pressure control achieved by benazepril (10 mg/day) and valsartan (80 mg/day) combined therapy with those achieved by benazepril (20 mg/day) or valsartan (160 mg/day) alone. RESULTS: Despite comparable changes in blood pressure and glomerular filtration rate (GFR), combined therapy decreased proteinuria more than benazepril (-56% vs. 45.9%, P=0.02) and valsartan (-41.5%, P=0.002). Changes in urinary protein to creatinine ratio followed the same trend. Filtration fraction and renal vascular resistances (RVR) decreased more with combined (-14.7%,-23.7%) or benazepril ( 12.4%, -20.5%) than with valsartan (-2.7%, -12.5%, P < 0.05 vs. both). RVR changes, adjusted for GFR changes, were associated with those in proteinuria (P < 0.05). Changes in glomerular permeability were comparable and did not predict different changes in proteinuria in the three groups. CONCLUSION: At comparable blood pressure, combined ACEi and ARA decreased proteinuria better than ACEi and ARA. The greater antiproteinuric effect most likely depended on an ACEi-related hemodynamic effect, in addition to glomerular size selectivity amelioration. Long term combined ACEi and ARA therapy may be more renoprotective than treatment with each agent alone. PMID- 12631094 TI - Role of endogenous nitric oxide in unilateral ureteropelvic junction obstruction in children. AB - BACKGROUND: Obstructive nephropathy leads to tubulointerstitial fibrosis and loss of renal function. Nitric oxide has been shown to have antifibrotic properties. We examined nitric oxide synthase (NOS) activity and expression in kidneys from children who underwent surgery release of unilateral ureteropelvic junction (UPJ) obstruction in relation to clinical and histologic parameters. METHODS: NOS activity and the expression of NOS isoforms measured at the mRNA level by reverse transcription-polymerase chain reaction (RT-PCR) assay were determined in tissue obtained by biopsy from obstructed kidneys of 18 children at the time of pyeloplasty. Tissue from kidneys removed because of various malignancies were issued as control. RESULTS: A significant increase in calcium/calmodulin independent NOS activity (iNOS) and iNOS mRNA expression was found in the medulla of obstructed kidneys. Calcium/calmodulin-dependent NOS activity (cNOS) and endothelial (eNOS) mRNA, by contrast, were increased in the cortex from obstructed kidneys. A role of tumor necrosis factor-alpha (TNF-alpha) on enhanced iNOS was suggested by the finding of increased urine levels in obstructed pelvis. Increased interstitium macrophage number, by immunolabeling of CD68, was related to the delay in obstruction release and to decreased glomerular filtration rate (GFR) at surgery. A positive linear relationship was found between cNOS activity in cortex and creatinine clearance. The degree of interstitial fibrosis correlated negatively with cNOS activity in cortex. CONCLUSION: In kidneys from children with UPJ obstruction an increased activity and expression of iNOS in medulla and cNOS-dependent eNOS in cortex were demonstrated. A role of cNOS in modulating GFR and interstitial fibrosis can be suggested. Prolonged UPJ obstruction would lead to a worsened prognosis on renal injury. PMID- 12631095 TI - Total hip bone mass predicts survival in chronic hemodialysis patients. AB - BACKGROUND: Bone mass measurements are widely used to diagnose osteoporosis and identify patients at risk for fractures. Recent studies have shown that reduced bone mass is also predictive of all-cause and cardiovascular mortality in healthy postmenopausal women. Among chronic hemodialysis patients reduced bone mass resulting from the combined effects of age-related factors and renal osteodystrophy has been associated with vascular calcification. In this prospective study, we investigated the ability of bone mass measurements to predict mortality in chronic hemodialysis patients. METHODS: Eighty eight patients underwent hip bone mass measurements by dual energy x-ray absorptiometry (DEXA) and were followed up for a mean of 3.5 years. Eleven patients received renal transplants and were therefore excluded. Forty of the remaining patients died, predominantly due to cardiovascular causes (43%). RESULT: Cox regression analysis showed patient age, osteoporosis or osteopenia, and baseline average calcium x phosphate product (Ca x P) to be independently predictive of increased mortality. Patients with osteopenia or osteoporosis had a 3.3- and 4.3-fold increased risk of death, respectively. A Ca x P of >or=5.0 mmol2/L2 predicted a threefold increased risk of death. Previous renal transplantation predicted a lower mortality. CONCLUSIONS: This study has shown for the first time that reduced total hip bone mass is an independent predictor of all-cause mortality among chronic hemodialysis patients. Our findings suggest that reduced bone mass and elevated Ca x P may be involved in the pathogenesis of cardiovascular disease in these patients but further research is required to investigate the possible mechanisms for these associations. PMID- 12631096 TI - Level of kidney function as a risk factor for cardiovascular outcomes in the elderly. AB - BACKGROUND: There is a high prevalence of both reduced kidney function as well as cardiovascular disease (CVD) in the elderly. We evaluated whether the level of kidney function is an independent risk factor for CVD outcomes in the Cardiovascular Health Study (CHS), a cohort of subjects whose age at baseline was 65 years old or older. METHODS: Cox proportional-hazards regression was used to evaluate the association of predicted glomerular filtration rate (GFR) with CVD after adjustment for the major CVD risk factors. We searched for nonlinear relationships between GFR and CVD, as well as interactions between level of kidney function and major CVD risk factors on CVD. RESULTS: A total of 4893 subjects with predicted GFR of 15 to 130 mL/min/1.73 m2 were included in the analysis. Fifty-six percent were female and the mean age was 73.4 years. Of the subjects, 549 (11.2%) died and 1229 (25.1%) experienced CVD events in 5.05 years of follow-up. Each 10 mL/min/1.73 m2 lower GFR was associated with an adjusted hazard ratio for CVD, de novo CVD, recurrent CVD and all-cause mortality of 1.05 (1.02, 1.09), 1.07 (1.01, 1.12), 1.04 (0.99, 1.09), and 1.06 (1.00, 1.12), respectively. There was no significant interaction between level of GFR and other traditional CVD risk factors on CVD outcomes. A linear model best described the relationship between GFR and CVD. CONCLUSION: The level of GFR is an independent risk factor for CVD, de novo CVD, and all-cause mortality in the elderly. PMID- 12631098 TI - Free cysteine is increased in plasma from hemodialysis patients. AB - BACKGROUND: Although only the total thiol concentration, which includes bound and free forms, has been determined in most previous clinical studies, the free form may be a better predictor of cardiovascular risk. METHODS: We measured the apparent concentration of free homocysteine (Hcy) and cysteine (Cys) in filtered and acid-soluble fractions of plasma in healthy control subjects and in patients with chronic renal failure just before and after a hemodialysis session. RESULTS: In control, filtered Hcy and acid-soluble Hcy were similar, while filtered Cys was much smaller than in acid-soluble Cys. In prehemodialysis samples, filtered Cys was more than 60 times as abundant (259.2 +/- 26.2 micromol/L) as in control samples (4.1 +/- 0.7 micromol/L ). Free-to-total ratios for filtered Cys were 1.6 +/- 0.3% in controls, but 40.9 +/- 2.7% in prehemodialysis patients. CONCLUSIONS: The filtered fraction of thiols can be used to estimate solute transport across the dialysis membrane. In addition, the possible involvement of cysteine in the pathogenesis of atherosclerosis in hemodialysis patients should be reexamined. PMID- 12631097 TI - Zoledronic acid to prevent bone loss in the first 6 months after renal transplantation. AB - BACKGROUND: Bisphosphonates can prevent bone mineral density loss after renal transplantation, but their effect on trabecular mineralization and bone morphology, two key factors of bone stability, remains unknown. METHODS: In a 6 month, randomized, placebo-controlled study, 20 kidney transplant recipients received either 4 mg zoledronic acid or placebo twice within 3 months after engraftment. At transplantation and after 6 months, mean trabecular calcium concentration and trabecular morphometry were measured in bone biopsies. Bone mineral density (BMD) of the femoral neck and the lumbar spine were evaluated by dual-energy x-ray absorptiometry, and serum biochemical markers of bone metabolism were determined monthly. RESULTS: Trabecular calcium content increased significantly in the zoledronic acid group, but remained unchanged in the placebo group. BMD at femoral neck showed no change in the zoledronic acid group, but decreased in the placebo group. BMD of the lumbar spine was increased in the zoledronic acid group without change in the placebo group. High-turnover bone disease resolved similarly in both groups, as evidenced by a significant decrease of eroded bone surface, osteoclast and osteoblast surface. Serologic markers of bone formation and resorption were significantly lower in zoledronic acid-treated patients throughout the study. Kidney transplant function was stable after zoledronic acid therapy. CONCLUSIONS: Our results show that administration of zoledronic acid improves the calcium content of cancellous bone after kidney transplantation. The beneficial effect of bisphosphonate therapy is further evidenced by an increase of lumbar spine BMD, and stabilization of femur BMD. PMID- 12631099 TI - An in vivo method for adenovirus-mediated transduction of thick ascending limbs. AB - BACKGROUND: The thick ascending limb of the loop of Henle (THAL) plays an important role in the maintenance of salt, water, and acid-base balance. While techniques for gene transfer of renal vascular cells and some tubular segments have been described, in vivo transduction of THALs has not been successful. We hypothesized that in vivo injection of adenoviral vectors into the renal medulla would result in efficient transduction of THALs. METHODS: We injected recombinant adenoviruses containing the reporter gene, green fluorescent protein (GFP), driven by either the cytomegalovirus promoter (Ad-CMVGFP) or the promoter for the Na/K/2 Cl cotransporter (Ad-NKCC2GFP), which is THAL-specific, into the outer medullary interstitium of Sprague-Dawley rat kidneys. Kidneys were removed at various times after viral injection and analyzed for GFP expression. RESULTS: Western blots revealed strong GFP expression in the outer medulla (which is composed primarily of THALs) 5 days after Ad-CMVGFP injection. We quantified THAL transduction efficiency by scoring the number of fluorescent tubules in THALs suspensions, which showed that at least 77 +/- 3% of THAL expressed GFP. To specifically transduce THALs, we injected Ad-NKCC2GFP into the medullary interstitium. As determined by Western blot, GFP expression was only detected in the outer medulla. Immunohistochemistry and confocal microscopy showed that GFP was localized to tubular cells positive for Tamm-Horsfall protein. Thus, GFP fluorescence was only detected in THALs, not in cortical, inner medulla or vascular cells. Time-course studies showed that GFP expression in THALs was measurable from 4 to 14 days, peaked at 7 days, and had returned to background levels by 21 days. CONCLUSION: This method facilitates highly efficient, THAL specific transduction. While application of this technique for gene therapy in humans is unlikely due to the transient gene expression observed and the impossibility for repeated injections of adenoviral vectors, this method provides a valuable tool for investigators studying regulation and mechanisms of THAL ion transport and its relationship to whole-kidney physiology and pathophysiology. PMID- 12631100 TI - Effect of a novel adsorbent on cytokine responsiveness to uremic plasma. AB - BACKGROUND: Middle molecules such as beta2-microglobulin (beta2M) and advanced glycation end products (AGE)-modified proteins contribute to inflammation in uremia. The BetaSorb column is a new adsorptive device, which contains copolymeric beads, suitable for removal of beta2M and other middle molecules. We assessed the effect of this column on the bioreactivity of uremic plasma, as measured by cytokine responsiveness. METHODS: Uremic plasma was perfused in vitro through the column (10 mL/min) and samples were collected after 10 to 30 passes. Endotoxin-stimulated tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) production by THP-1-derived monocytes was measured following brief exposure to uremic plasma. beta2M levels were measured. The contribution of AGE modified proteins to the bioreactivity of uremic plasma was explored. RESULTS: TNF-alpha and IL-10 production markedly decreased after 30 passes (629 +/- 78 vs. 144 +/- 62 pg/mL; 207 +/- 25 vs. 117 +/- 23 pg/mL; P=0.04). The column removed beta2M efficiently with a marked decline in plasma levels by 99% after 30 passes. Neutralization of AGE receptor (RAGE) resulted in a further reduction in the bioreactivity of uremic plasma. This was observed with nonperfused, as well as perfused, uremic plasma, suggesting that AGE-modified proteins were biologically active and still present after perfusion. CONCLUSION: The sorbent beads removed uremic solute(s) that prime monocytes to enhanced cytokine production. Removal of beta2M was efficient, and of native and AGE-modified middle molecules likely. PMID- 12631101 TI - Muscle crush compartment syndrome: fulminant local edema with threatening systemic effects. PMID- 12631102 TI - Of laminins and delamination in Alport syndrome. PMID- 12631103 TI - A refined understanding of immunosuppressives and cancer risk. PMID- 12631104 TI - Hyaline arteriolosclerosis: new meaning for an old lesion. PMID- 12631105 TI - Rapidly progressive crescentic glomerulonephritis. PMID- 12631106 TI - Chemokines and chemokine receptors are involved in the resolution or progression of renal disease. AB - Locally secreted chemokines mediate leukocyte recruitment during the initiation and amplification phase of renal inflammation. In turn, the infiltrating leukocytes contribute to renal damage by releasing inflammatory and profibrotic factors. Rapid down modulation of the chemokine signal will support resolution of acute inflammation, whereas progression occurs if ongoing or repeated renal injury maintains continuous local chemokine secretion and leukocyte influx into the glomerulus or the interstitial space. In glomerular injury proteinuria itself as well as glomerular secreted cytokines stimulate downstream tubular epithelial cells to also secrete chemokines. During primary tubular injury, tubular epithelial cells directly become a major site of chemokine production. This in turn supports leukocyte infiltration and activation. Infiltrating leukocytes stimulate fibroblast proliferation and matrix synthesis, leading to widening of the interstitial space. The specific and intricate renal vascular architecture renders the organ susceptible to ischemic damage as interstitial volume increases. Ischemia in turn serves as a stimulus for chemokine and cytokine production and matrix synthesis. The mutual stimulation between fibroblasts and infiltrating leukocytes supports progressive tubular damage, renal fibrosis, and glomerulosclerosis. Potentially this vicious circle leading to progression of chronic nephropathies offers the opportunity for therapeutic intervention. Interfering with the chemokine network that mediates leukocyte recruitment may represent a promising therapeutic option for progressive renal disorders and renal fibrosis. This article summarizes the present data on the role of chemokines in acute and chronic renal disease with special emphasis on their potential role in mediating resolution or progression of renal disease as well as on therapeutic options. PMID- 12631107 TI - Calcitonin receptor isoforms expressed in the developing rat kidney. AB - BACKGROUND: Development in the metanephric-kidney transition period involves the precise expression of paracrine and autocrine events in an ordered spatio temporal manner. Expression of these molecular events is tightly controlled and includes positive and negative growth factors and cognate receptors within close proximity in developing structures in the expanding renal cortex and medulla. The expression of calcitonin receptor (CTR) isoforms C1a and C1b in this context has not previously been described. Our current study also explored the relationship between the expression of CTR isoforms and amylin binding sites. METHODS: Techniques included immunohistochemistry with novel antibodies that detect CTR isoforms, real time PCR for the quantification of CTR isoforms, Western blot and in vitro autoradiography, on tissues from embryo day 18 to postnatal day 30. RESULTS: The CTR C1a isoform is expressed in the ureteric ducts of the metanephros and both isoforms are expressed in the developing distal convoluted tubules, ascending limbs of the loop of Henle and collecting ducts in the postnatal rat kidney. There was a 60-fold excess of C1a versus C1b isoforms. An apparent molecular weight of 63 kD was found. In vitro autoradiography demonstrated that while amylin binding sites were predominantly in the cortex, CTR expression was largely localized in the medulla in an earlier event, followed by cortical expression. CONCLUSIONS: CTR C1a protein expression has been identified in the ureteric ducts in the metanephros and both isoforms expressed in the distal portions of the developing nephrons and collecting ducts. Since amylin binding sites have been localized on the proximal tubules of the cortex, it is unlikely that amylin receptors can be represented by modification of CTR affinity with receptor activity modifying proteins in the kidney. PMID- 12631108 TI - Renal activation of extracellular signal-regulated kinase in rats with autosomal dominant polycystic kidney disease. AB - BACKGROUND: Abnormal proliferation of renal tubule epithelial cells is a central factor in the biogenesis and sustained expansion of cysts in autosomal-dominant polycystic kidney disease (ADPKD). Recent evidence from in vitro studies of human cyst wall epithelial cells has implicated a role for the mitogen-activated protein (MAP) kinase pathway in this aberrant proliferation. To determine the extent to which this signaling pathway is involved in cyst pathogenesis in vivo, we measured the expression of select components of the MAP kinase cascade in Han:SPRD rats with ADPKD at an early stage of the disease. METHODS: Kidneys of 8 week-old normal Han:SPRD rats (+/+) or rats heterozygous (Cy/+) for ADPKD were examined by Western blot analysis and immunohistochemistry to determine the expression of extracellular-regulated kinase (ERK), phosphorylated ERK (P-ERK), Raf-1 (MAPKKK), phosphorylated Raf-1 (P-Raf-1), B-Raf, Rap-1 and phosphorylated protein kinase A (P-PKA). RESULTS: P-ERK was expressed to a greater extent in Cy/+ kidneys (3.74 +/- 1.07 fold) than in normal kidneys, whereas ERK abundance was not different. P-Raf-1 levels were higher in Cy/+ than in +/+ kidneys (1.53 +/- 0.08 fold) consistent with upstream stimulation of receptor tyrosine kinase. B-Raf and Raf-1 abundances were greater in Cy/+ than in +/+ (1.74 +/- 0.25 and 1.27 +/- 0.08 fold, respectively). In Cy/+, immunohistochemistry showed increased P-ERK and B-Raf expression in the abnormal mural epithelial cells within cysts. These findings, together with the detection of P-PKA and the small G protein, Rap 1, in cyst epithelial cells, implicate a potential role for cyclic adenosine monophosphate (AMP) in the activation of ERK in ADPKD cells. CONCLUSIONS: We conclude that the MAP kinase pathway is activated to the level of ERK in the abnormal mural epithelial cells lining cysts in animals with a dominantly inherited type of polycystic kidney disease. We suggest that cAMP, acting through PKA, Rap-1 and B-Raf, may contribute to the activation of ERK in a way that complements receptor tyrosine kinase-mediated agonists in the promotion of cyst enlargement. PMID- 12631109 TI - Preemptive ramipril therapy delays renal failure and reduces renal fibrosis in COL4A3-knockout mice with Alport syndrome. AB - BACKGROUND: Alport syndrome (AS) is a common hereditary cause of end-stage renal failure in adolescence due to defects in type IV collagen genes. Molecular genetics allows early diagnosis, however, no preventive strategy can be offered. Using the COL4A3 -/- mouse, an animal model for human AS, we evaluated therapy with ramipril in mice. METHODS: One hundred and twenty-two Alport-mice were treated with 10 mg/kg/day ramipril added to drinking water. Proteinuria, serum urea and lifespan were monitored. Renal matrix was characterized by immunohistochemistry, light- and electron microscopy, and Western blot. RESULTS: Untreated COL4A3 -/- mice died from renal failure after 71 +/- 6 days. Early therapy starting at four weeks of age and continuing to death delayed onset and reduced the extent of proteinuria. Uremia was postponed by three weeks in treated animals. Lifespan increased by more than 100% to 150 +/- 21 days (P < 0.01). In parallel, decreased deposition of extracellular matrix and lessened interstitial fibrosis as well as reduced amounts of renal transforming growth factor-beta1 (TGF-beta1) could be demonstrated. Late therapy starting at seven weeks decreased proteinuria, however, lifespan did not increase significantly. CONCLUSIONS: The results indicate an antiproteinuric and antifibrotic nephroprotective effect of ramipril in COL4A3 -/- mice is mediated by down-regulation of TGF-beta1. This effect in mice is enhanced by initiation of therapy during pre-symptomatic disease. The data in COL4A3 -/- mice as an animal-model for Alport syndrome suggest that ramipril might as well delay renal failure in humans with AS. Early diagnosis and preemptive treatment also may be crucial in humans. PMID- 12631110 TI - Mutations in the COL4A4 gene in thin basement membrane disease. AB - BACKGROUND: Patients with thin basement membrane disease (TBMD) are often from families where hematuria segregates with the COL4A3 and COL4A4 genes. These genes also are affected in autosomal recessive Alport syndrome. The aim of this study was to demonstrate COL4A4 mutations in TBMD. METHODS: Forty-eight unrelated individuals with TBMD who had no family members with autosomal recessive Alport syndrome were examined for COL4A4 mutations. The diagnosis of TBMD had been confirmed by renal biopsy (43/48, 90%) or by a family history of hematuria but without a renal biopsy (5/48, 10%). The 47 coding exons of COL4A4 were screened for mutations with the methods of enzyme mismatch cleavage or single stranded conformational polymorphism (SSCP) analysis, and exons that demonstrated electrophoretic abnormalities were sequenced. RESULTS: Nine variants that altered the coding sequences were identified. These were nonsense and frameshift mutations that resulted in stop codons (N = 3), and glycine (N = 3) and non glycine missense variants (N = 3). Four intronic variants and three neutral polymorphisms were also detected. In total, four variants were considered 'pathogenic' principally because they resulted in stop codons or were not present in non-hematuric normal subjects. Three variants were considered 'possibly pathogenic' but two of these were each present in one of 46 non-hematuric normal subjects. CONCLUSIONS: Pathogenic COL4A4 mutations were demonstrated in three of the nine (33%) families in whom hematuria segregated with the COL4A3/COL4A4 locus. Two stop codons (R1377X and 2788/91delG) and a glycine substitution (G960R) resulted in hematuria in all 16 members who were tested from these three families. The S969X mutation described here in TBMD for the first time, as well as the R1377X mutation, also occur in autosomal recessive Alport syndrome. PMID- 12631111 TI - Long-term treatment with ramipril attenuates renal osteopontin expression in diabetic rats. AB - BACKGROUND: Osteopontin (OPN) mediates progressive renal injury in various renal diseases by attracting macrophages, and its expression is regulated by the renin angiotensin system (RAS). We studied the association between OPN expression and tubulointerstitial injury, and investigated the effect of ramipril on OPN expression in an animal model of non-insulin-dependent diabetes mellitus (NIDDM): Otsuka Long-Evans Tokushima Fatty (OLETF) rats. METHODS: Control (Long-Evans Tokushima Otsuka, LETO) and diabetic (OLETF) rats were treated with ramipril (3 mg/kg in drinking water) or vehicle for nine months, starting at 20 weeks of age. Systolic blood pressure, body weight, urinary protein excretion and oral glucose tolerance tests (OGTT) were monitored periodically. Renal function, histology (glomerulosclerosis, tubulointerstitial fibrosis, and ED-1-positive cells as a measure of macrophage infiltration), and expressions of OPN and transforming growth factor-beta1 (TGF-beta1) were evaluated at the end of the study. RESULTS: Compared with the LETO rats, OLETF rats showed declines in creatinine clearance rate, increases in urinary protein excretion and systolic blood pressure, and development of glomerulosclerosis, tubulointerstitial fibrosis, and inflammatory cell infiltration (all P < 0.05). Blocking angiotensin II with ramipril significantly improved all of these parameters (all P < 0.01). At the molecular level, expressions of OPN and TGF-beta1 were up-regulated in the OLETF rats, and were markedly suppressed following ramipril treatment. The sites of strong OPN mRNA and protein expressions were localized to areas of renal injury. Of note, the expression of OPN mRNA was strongly correlated with the number of ED-1 positive cells (r = 0.560, P = 0.01) and the tubulointerstitial fibrosis score (r = 0.500, P < 0.05). CONCLUSIONS: Up-regulation of OPN expression may play a role in tubulointerstitial injury associated with diabetic nephropathy, and blockade of the RAS by ramipril may confer renoprotection by decreasing OPN expression in non-insulin-dependent diabetic nephropathy. PMID- 12631112 TI - Advanced glycation end products inhibit de novo protein synthesis and induce TGF beta overexpression in proximal tubular cells. AB - BACKGROUND: We have shown previously that OPB-9195, a novel inhibitor of advanced glycation end products (AGE), significantly prevented renal tubular injury and tubulointerstitial fibrosis in spontaneous diabetic rats. However, the molecular mechanisms underlying this have not been fully elucidated. METHODS: Three immunochemically distinct AGE were prepared by incubating bovine serum albumin (BSA) with glucose, glyceraldehyde, or methylglyoxal. Then, the effects of AGE on human proximal tubular epithelial cells were examined. The intracellular formation of reactive oxygen species (ROS) was detected using the fluorescent probe CM-H2DCFDA. DNA synthesis was evaluated by thymidine uptake, and de novo protein synthesis was determined by [3H]leucine incorporation. Prostaglandin E2 (PGE2) and transforming growth factor-beta (TGF-beta) released into media were quantitatively analyzed in an enzyme-linked immunosorbent assay. TGF-beta gene expression was analyzed by quantitative reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: When these AGE-BSA were administered to tubular cells, each of them increased generation of intracellular ROS. All of the AGE BSA, but not non-glycated BSA, were found to induce statistically significant decreases in de novo protein synthesis and PGE2 secretion by tubular cells. Furthermore, AGE-BSA up-regulated the levels of mRNAs for TGF-beta in tubular cells. The structural epitope designated glucose-derived AGE was found to have the greatest cytopathic effects on tubular cells. These AGE-induced inhibition of protein synthesis and PGE2 secretion as well as the up-regulation of TGF-beta mRNA were found to be completely prevented by N-acetylcysteine. Furthermore, H2O2 was shown to inhibit protein synthesis and PGE2 secretion by proximal tubular cells in a dose-dependent manner. CONCLUSION: The results suggest that AGE inhibits de novo protein synthesis and stimulates TGF-beta mRNA expression in proximal tubular epithelial cells through overgeneration of intracellular ROS. Thus, AGE are involved in the pathogenesis of tubular injury in diabetic nephropathy. PMID- 12631113 TI - Tumor necrosis factor-alpha stimulates fractalkine production by mesangial cells and regulates monocyte transmigration: down-regulation by cAMP. AB - BACKGROUND: Fractalkine is a CX3C chemokine for mononuclear cells that has been implicated in the recruitment and accumulation of monocytes seen in glomerular diseases. We investigated the mechanisms by which tumor necrosis factor (TNF) alpha stimulates mesangial cell (MC) fractalkine expression, and the effects of MC-derived fractalkine on monocyte transmigration. METHODS: Cultured rat MCs were incubated with TNF-alpha, with or without pretreatment with pharmacologic inhibitors of protein kinases or transcriptional factors downstream to TNF-alpha. Fractalkine mRNA and protein were analyzed by Northern and Western blotting. Translocation of nuclear factor (NF)-kappaB was evaluated by immunocytochemical staining. Monocyte transmigration was determined by in vitro chemotaxis assay. RESULTS: TNF-alpha stimulated MC fractalkine mRNA as well as cell-bound and soluble protein expression in a dose- and time-dependent manner. The soluble fractalkine was shed from the cell-bound form via metalloproteinase-dependent cleavage, and mediated in part TNF-alpha-induced monocyte transmigration in vitro. The incubation of MCs with calphostin C [a selective inhibitor of protein kinase C (PKC)] or PD98059 [a selective inhibitor of p42/44 mitogen-activated protein kinase (MAPK) kinase] attenuated TNF-alpha-stimulated fractalkine mRNA and protein expression. Coincubation of MCs with calphostin C and PD98059 resulted in a synergistic inhibition of TNF-alpha-stimulated fractalkine mRNA and protein expression. Incubation of MCs with phorbol myristate acetate (PMA) for four hours resulted in an increase in fractalkine mRNA expression that could be suppressed by calphostin C or depletion of PKC by pretreatment with PMA for 24 hours. Further, activation of PKC-depleted MCs with TNF-alpha stimulated fractalkine mRNA expression that could be blocked by calphostin C. PD 98059, but not calphostin C, inhibited TNF-alpha-activated phospho-p42/44 MAPK and phospho-c Jun levels, whereas only calphostin C inhibited TNF-alpha-activated phosphorylation of PKCzeta/iota. The incubation of MCs with MG132, a NF-kappaB inhibitor, abolished TNF-alpha-induced degradation of inhibitory protein of NF kappaB (I-kappaB)alpha, nuclear translocation of NF-kappaB, and fractalkine expression, without affecting phospho-c-Jun levels. In contrast, curcumin, an activating protein (AP)-1 inhibitor, attenuated TNF-alpha-stimulated phospho-c Jun levels and fractalkine expression without discernible effects on TNF-alpha induced degradation of I-kappaBalpha or NF-kappaB nuclear translocation. Neither PD 98059 nor calphostin C affected TNF-alpha-induced degradation of I-kappaBalpha or NF-kappaB nuclear translocation. Additional experiments examining the role of cAMP on MC fractalkine expression showed that the incubation of MCs with TNF alpha and either db-cAMP or forskolin attenuated TNF-alpha-stimulated fractalkine mRNA and protein expression, preceded by attenuation of TNF-alpha-activated phosphorylation of p42/44 MAPK, and c-Jun, but not phosphorylation of PKCzeta/iota or nuclear translocation of NF-kappaB. CONCLUSION: The present data indicate that TNF-alpha activation of PKCzeta/iota, p42/44 MAPK, c-Jun/AP-1, and p65/NF-kappaB are involved in TNF-alpha-stimulated MC fractalkine expression, with the soluble fractalkine mediating in part the TNF-alpha-induced monocyte transmigration in vitro. Uncoupling of p42/44 MAPK or c-Jun/AP-1 signals may contribute to cAMP inhibition of MC fractalkine expression activated by TNF alpha. PMID- 12631114 TI - PTH ameliorates acidosis-induced adverse effects in skeletal growth centers: the PTH-IGF-I axis. AB - BACKGROUND: Chronic metabolic acidosis (CMA) exerts profound adverse effects on bone metabolism thereby leading to impaired skeletal linear growth. We have recently shown that CMA in vitro causes distinct morphological changes in skeletal growth centers along with inhibition of endochondral differentiation. In addition, CMA causes an end organ resistance to the anabolic effects of growth hormone (GH) and locally produced insulin-like growth factor-I (IGF-I) in skeletal growth centers. Given the effects of parathyroid hormone (PTH) and PTH related protein (PTHrP) on the development of cartilaginous bone, we sought to determine whether PTH has any effects on the changes induced by CMA in skeletal growth centers. The interaction between PTH and IGF-I in growth centers during neutral or acidic conditions were studied specifically. METHODS: An in vitro organ culture system using the murine mandibular condyle was employed as a model for endochondral active growth center. Condyles from six-day-old mice were cultured in BGJb medium of either neutral pH (pH approximately 7.4) or acidic pH (pH approximately 7.15) in the presence or absence of 10-10 mol/L [1-34] PTH. After 24, 48, 72 and 96 hours of culture, the condyles were washed, fixed in formaldehyde, and processed for paraffin embedding. Histologic markers of the growth center were assessed. In addition, the protein level and mRNA expression for various markers of cartilage differentiation were evaluated by immunohistochemistry and in situ hybridization, respectively. The abundance and expression levels of IGF-I and IGF-I receptor (IGF-I-R) were assessed also. RESULTS: Following incubation for 72 hours in acidic conditions, there was a marked attenuation of the chondroblastic zone, suggesting a defect in the process of cellular differentiation. Acidosis also down-regulated endochondral differentiation markers (cartilage specific proteoglycans, collagen type II). This was accompanied by a reduction in the expression of IGF-1, IGF-1 receptor and PTH receptors. PTH (10-10 mol/L) added to acidic cultures prevented the adverse effects of CMA on endochondral differentiation and increased the overall condylar growth, when compared to acidic conditions without PTH. PTH also up regulated its own receptor in control as well as during acidic conditions, and increased the expression levels of IGF-1 and IGF-1 receptor in the acidotic condyle. Acidosis increased the expression of IGF-I binding protein-4 (IGFBP-4, an inhibitor of IGF-I activity), whereas coincubation with PTH during acidic conditions abrogated the up-regulation of IGFBP-4. Addition of a neutralizing antibody to IGF-I-R during PTH treatment under acidic conditions resulted in the abrogation of the ameliorative effect of PTH on endochondral differentiation. The protein kinase C (PKC) signaling pathway was modulated negatively by CMA. However, PTH activated PKC-alpha under both control and acidic conditions. The phorbol ester, PMA (phorbol 12-myristate 13-acetate), a PKC activator, mimicked the effect of PTH on chondrocyte differentiation. CONCLUSION: Parathyroid hormone at low concentration stimulates the differentiation and proliferation of cartilage cells and prevents the suppressive effect of acidosis on endochondral bone differentiation and on the IGF-I/IGF-I-R system in skeletal growth centers. Increased local production of IGF-I by PTH, which takes place even during acidotic conditions, mediates, at least in part, the ameliorative effect of PTH. Protein kinase C is probably one of the signaling pathways mediating the salutary effects of PTH on chondrocyte differentiation in growth centers. This study lends further credence to the notion that under certain conditions, PTH or PTHrP can exert anabolic effects in the skeleton. These findings may be of clinical therapeutic significance in children and patients with CMA. PMID- 12631115 TI - Gene transfer of truncated IkappaBalpha prevents tubulointerstitial injury. AB - BACKGROUND: Severe proteinuria not only indicates the presence of progressive glomerular disease, but also causes tubular epithelial cells to produce inflammatory mediators leading to tubulointerstitial (TI) injury. We investigated the role of nuclear factor-kappaB (NF-kappaB) in tubular epithelial cells in the development of proteinuria-induced TI injury. METHODS: To specifically inhibit NF kappaB activation, a recombinant adenovirus vector expressing a truncated form of IkappaBalpha (AdexIkappaBDeltaN) was injected into renal arteries of protein overloaded rats, a model of TI injury characterized by infiltration of mononuclear cells and fibrosis. RESULTS: Activation of NF-kappaB in the renal cortex, observed in protein-overloaded rats treated with a control vector, recombinant lacZ adenovirus, was prevented in AdexIkappaBDeltaN-injected rats. Microscopic examination revealed AdexIkappaBDeltaN treatment to markedly attenuate proteinuria-induced TI injury. Increased immunostaining of vascular cell adhesion molecule-1, transforming growth factor-beta, and fibronectin in TI lesions also was suppressed by AdexIkappaBDeltaN injection. CONCLUSIONS: These findings provide evidence of the critical role of NF-kappaB activation in TI injury and suggest the therapeutic potential of adenovirus-mediated IkappaBDeltaN gene transfer into the kidney as a means of interrupting the process of TI damage. PMID- 12631116 TI - Complexes of IgA with FcalphaRI/CD89 are not specific for primary IgA nephropathy. AB - BACKGROUND: The presence of IgA together with the myeloid IgA-receptor FcalphaRI/CD89 in the circulation of patients with IgA nephropathy (IgAN) has been suggested as a specific pathogenic factor for mesangial deposition. However, in a recent study we found these complexes also in serum samples from healthy subjects. To investigate whether these circulating complexes are specific for IgAN, the levels and characteristics of IgA-CD89 complexes were analyzed in patients with IgAN and healthy controls. METHODS: Specific ELISAs with different poly- and monoclonal antibodies and a sensitive dot-blot method were used to measure IgA-CD89 levels in serum and purified IgA samples obtained from healthy volunteers (N = 30) and patients with IgAN (N = 35). Fractionated samples of purified IgA were used to compare the size characteristics of the IgA-CD89 complexes. RESULTS: Almost all CD89 in serum of patients with IgAN and controls was associated with high molecular weight IgA. Quantitative analysis of IgA-CD89 complexes in purified IgA revealed no significant difference between patients with IgAN and controls. No correlation was found between levels of IgA-CD89 complexes and clinical parameters associated with progressive IgAN. CONCLUSIONS: CD89 in the circulation is found mainly linked to high molecular weight IgA. The presence of these complexes is not specific for IgAN. Therefore, if IgA-CD89 complexes are involved in the pathogenesis of primary IgA nephropathy, additional factors are required to explain the IgA-CD89 complex-mediated renal inflammation. PMID- 12631117 TI - Nitric oxide modulates vascular endothelial growth factor and receptors in chronic cyclosporine nephrotoxicity. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is involved in angiogenesis, wound healing, and inflammation and exerts its effect via tyrosine kinase receptors, fms-like tyrosine kinase (Flt-1) and fetal liver kinase (Flk-1 or KDR). We have previously shown that VEGF is up-regulated in a model of chronic cyclosporine (CsA) nephrotoxicity and that l-arginine (l-Arg) improved while N nitro-l-arginine-methyl ester (L-NAME) worsened fibrosis. We examined the role of nitric oxide modulation on VEGF in this model. METHODS: Pair-fed salt-depleted rats were administered CsA, CsA + L-NAME, CsA +l-Arg, vehicle (VH), VH + L-NAME or VH +l-Arg and were sacrificed at 7 or 28 days. Physiologic and histologic changes were studied in addition to the mRNA expression of VEGF and its receptors Flt-1 and KDR/Flk-1 by Northern blot and the protein expression of VEGF by Western blot and immunohistochemical staining. RESULTS: While L-NAME worsened renal function and histology, l-Arg had the opposite beneficial effect in CsA treated rats. VEGF mRNA and protein expressions increased with CsA, further increased with L-NAME and became significantly reduced with L-Arg. Flt-1 expression was similar in all groups. On the other hand, KDR/Flk-1 mRNA expression was modulated in a fashion similar to VEGF. Also, nitric oxide modulation did not have an effect on VH-treated rats. CONCLUSIONS: VEGF expression in chronic CsA nephrotoxicity is increased by nitric oxide blockade and decreased by nitric oxide enhancement. Moreover, VEGF probably exerted its effect via the KDR/Flk-1 receptor. The actions of VEGF in this model remain speculative, but it is probable that VEGF plays a role, either independently or through nitric oxide, in CsA-induced fibrosis. PMID- 12631119 TI - Reduced postischemic macrophage infiltration and interstitial fibrosis in osteopontin knockout mice. AB - BACKGROUND: Osteopontin (OPN) is a phosphoprotein that is up-regulated in several experimental models of renal disease, including ischemia/reperfusion injury. OPN has been described as a macrophage chemoattractant, may serve as a survival factor for tubular cells, and is implicated in the development of tubulointerstitial fibrosis. However, the precise role of this protein in renal pathophysiology remains unclear. METHODS: OPN knockout and wild-type mice were subjected to 30 minutes of warm renal ischemia combined with a contralateral nephrectomy, and sacrificed at six different time points, ranging from 12 hours to seven days after reperfusion. Besides functional and morphological parameters of postischemic acute renal failure (ARF), macrophage infiltration, apoptosis and expression of collagen types I and IV were investigated. RESULTS: Postischemic ARF in OPN knockouts and wild-types showed a similar course and severity, without significant differences in either functional or morphological disease parameters. However, macrophage infiltration was significantly diminished in OPN knockouts after five and seven days, in cortex as well as in the outer stripe of the outer medulla (OSOM). Furthermore, OPN knockout mice showed significantly enhanced apoptosis in the injury phase and significantly less collagen I and IV expression in the regeneration phase of postischemic ARF. CONCLUSIONS: There was no influence of OPN protein on the severity or course of functional impairment or morphological injury in the first seven days after an ischemic insult to the kidney. However, our results demonstrate that OPN favors macrophage recruitment to the postischemic kidney, inhibits apoptosis, and stimulates the development of renal fibrosis after an acute ischemic insult. PMID- 12631118 TI - Induction of DNA synthesis by ligation of the CD53 tetraspanin antigen in primary cultures of mesangial cells. AB - BACKGROUND: The interaction of mesangial cells with the extracellular matrix plays a major role in kidney biology. Tetraspanin proteins modulate cell interaction with the extracellular matrix. Tetraspanins form supramolecular structures on the cell membrane that send signals after engagement by unknown ligands, modulate different signaling processes, and regulate cell adhesion and motility. METHODS: CD53 was determined by immunohistochemistry, and on the cell surface of cultured rat mesangial cells by flow cytometry. Mesangial cell cultures were stimulated with MRC OX-44 antibody. DNA synthesis was measured by thymidine incorporation. Extracellular signal-regulated kinase (ERK) activation was determined by Western blot. RESULTS: CD53 was present in mesangial cells in vivo and in culture. Ligation of CD53 antigen with a monoclonal antibody triggered the induction of DNA synthesis, which was not sensitive to inhibitors of signaling pathways that use phosphatidylinositol 3-kinase (PI3K) and protein kinase C, or to calcium channel inhibitors, such as thapsigargin and verapamil. The DNA synthesis was inhibited by PD98059, a specific inhibitor of MEK that prevents ERK1/ERK2 activation. In addition, ERK1 and ERK2 activation by phosphorylation occurred following CD53 antigen ligation. The DNA synthesis was due to de novo synthesis and not to DNA repair as a consequence of the initiation of apoptosis, determined by flow cytometry, and lack of proteolytic activation of PARP by caspase 3. CD53 antigen ligation also induced an increase in mitochondrial activity. CONCLUSIONS: To our knowledge this is the first identification of a tetraspanin protein in mesangial cells. CD53 antigen delivers a signal that initiates DNA synthesis. This signal is mediated by ERK1/ERK2 activation, but it is not sufficient to complete the cell cycle. PMID- 12631120 TI - Bioflavonoids attenuate renal proximal tubular cell injury during cold preservation in Euro-Collins and University of Wisconsin solutions. AB - BACKGROUND: Cold ischemia and reperfusion during kidney transplantation are associated with release of free oxygen radicals and damage of renal tubular cells. Bioflavonoids may diminish cold storage-induced injury due to antioxidant and iron chelating activities. This study was designed to delineate the renoprotective mechanisms of bioflavonoids and to define the structural features conferring cytoprotection from cold injury. METHODS: LLC-PK1 cells were preincubated for three hours with bioflavonoids and cold stored in University of Wisconsin (UW)- or Euro-Collins (EC)-solution for 20 hours. After rewarming, cell viability was assessed by the lactate dehydrogenase (LDH) release, MTT-test, and amino acid transport activity. Lipid peroxidation was assessed from the generation of thiobarbituric acid-reactive substances. RESULTS: Twenty-hours of cold storage of LLC-PK1 cells resulted in a substantial loss of cell integrity that was more pronounced in the EC (LDH release, 93.6 +/- 1.6%) than the UW solution (67.2 +/- 6.9%; P < 0.0001). Pretreatment with quercetin significantly enhanced cell survival (LDH release, 5.4 +/- 2.7% for UW and 8.4 +/- 4.2% for EC) in a concentration dependent manner. Structure-activity studies revealed similar renoprotection for kaempferol, luteolin and fisetin, unlike myricetin, morin, apigenin, naringenin, catechin, silibinin and rutin. Lipid peroxidation was reduced (UW alone, 2.7 +/- 1.2 vs. UW+quercetin 0.5 +/- 0.2 nmol/mg protein, P < 0.01), and l-threonine uptake completely sustained by pretreatment with quercetin, kaempferol, luteolin, and fisetin. However, renoprotection by fisetin was rapidly lost during rewarming. Protective properties of bioflavonoids were governed by the number and arrangement of hydroxyl substitutes, electron delocalization, sterical planarity, and lipophilicity of the basic diphenylpyran skeleton. CONCLUSION: Cold storage-induced renal tubular cell injury is ameliorated by bioflavonoids. Renoprotective effects of bioflavonoids are defined by structure, suggesting that flavonoids are incorporated into membrane lipid bilayers and interfere with membrane lipid peroxidation. PMID- 12631121 TI - Ureteral obstruction in neonatal mice elicits segment-specific tubular cell responses leading to nephron loss. AB - BACKGROUND: To elucidate the sequence of renal responses leading to nephron loss in obstructive nephropathy, we examined the evolution of segmental nephron cellular changes consequent to chronic unilateral ureteral obstruction (UUO) in the neonatal mouse. METHODS: Neonatal mice were subjected to UUO or sham operation, and kidneys were harvested 5, 12 or 19 days after surgery. Proximal tubules (PT), distal tubules (DT) and collecting ducts (CD) were identified with lectins. Histomorphometric quantitation was made for cellular necrosis, apoptosis, proliferation, tubular dilatation, tubular basement membrane (TBM) thickening, interstitial collagen, and glomerular maturation. The distribution of hypoxic tissue was determined using pimonidazole as a marker. Additional studies were performed by mechanically stretching monolayer cultures of mouse proximal tubular and collecting duct cells, and measuring apoptosis. RESULTS: Neonatal UUO induced an arrest of glomerular maturation throughout the period of study. Chronic UUO induced hypoxia, tubular necrosis, proliferation, and TBM thickening in the PT, but stimulated apoptosis in the DT and CD. Tubular dilation in the obstructed kidney was most severe in CD and least severe in PT. Tubular cell apoptosis closely paralleled tubular dilation (P < 0.05), and fibrosis surrounding individual tubules also correlated with tubular dilation (P < 0.001). Mechanical stretching of cultured mouse tubular cells induced apoptosis directly proportional to the magnitude of axial strain: apoptosis was consistently greater in CD than in PT cells (P < 0.05). CONCLUSIONS: Following UUO, the co localization of hypoxia with cellular proliferation, necrosis, and TBM thickening of the PT is consistent with ischemic injury resulting from vasoconstriction. In contrast, a selective dilation of the distal portion of the nephron (DT and CD), which results from the greater tubular compliance there, leads to stretch-induced epithelial cell apoptosis, along with a progressive peritubular fibrosis. Nephron loss in the obstructed developing kidney likely results from complex, segment specific cellular responses. PMID- 12631122 TI - Growth factor expression in a murine model of cryoglobulinemia. AB - BACKGROUND: Increased expression of growth factors including platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF-beta) are thought to play pivotal roles during mesangial expansion and glomerulosclerosis. Thymic stromal lymphopoietin (TSLP) transgenic mice develop mixed cryoglobulinemia and a membranoproliferative glomerulonephritis (MPGN). Here we describe the renal expression of isoforms of PDGF and TGF-beta in relation to changes in extracellular matrix (ECM) components and markers of cell proliferation and activation in this model. METHODS: A total of 123 mice, including 61 TSLP transgenic mice and 62 wild-type controls, were sacrificed at defined intervals. PDGF-A chain, -B chain, PDGF alpha- and beta-receptor (beta-R) and TGF-beta1 mRNA were analyzed by in situ hybridization. Expression of alpha smooth muscle actin (alphaSMA), collagen type I, collagen type IV, laminin, and a marker of proliferating cells (PCNA) were assessed by immunohistochemistry. Slides also were studied by combined immunohistochemistry and in situ hybridization with an antibody that recognizes monocytes/macrophage and with riboprobes that detect PDGF B-chain, PDGF beta-R or TGF-beta1 mRNA. RESULTS: Increased numbers of proliferating glomerular cells appeared early in the disease course, associated with de novo expression of alphaSMA. Expression of PDGF B-chain and beta-R mRNA was increased in the mesangium and in parietal epithelial cells of TSLP transgenic mice and correlated with the number of PCNA positive cells. Increased TGF-beta1 mRNA expression paralleled the deposition of type IV collagen. A significant proportion of Mac-2 positive macrophages expressed TGF-beta1 mRNA, while only a small percentage of glomerular macrophages expressed PDGF B-chain mRNA. No PDGF beta-R mRNA expression by macrophages was detected. CONCLUSION: TSLP transgenic mice develop a membranoproliferative glomerulonephritis in which glomerular cell proliferation and matrix deposition are associated with an increased expression of PDGF B-chain, PDGF beta-R and TGF-beta1. These findings extend the paradigms covering these growth factors established in the rat Thy 1 model of mesangiolysis and repairs to a murine model of progressive glomerulonephritis closely resembling human MPGN. PMID- 12631123 TI - A novel mechanism of nephron loss in a murine model of crescentic glomerulonephritis. AB - BACKGROUND: Nephron loss is a major determinant of renal failure in glomerular diseases. The prevalent concept stresses the role of the toxicity of filtered proteins and/or of interstitial inflammation in tubular degeneration. However, whether that concept is compatible with the actual histopathological features of nephron loss has not been investigated specifically. METHODS: We investigated the morphological aspects of tubular degeneration in crescentic glomerulonephritis in mice. Glomerulonephritis was induced by intravenous injection of anti-glomerular basement membrane antiserum in presensitized mice. Kidneys were fixed by perfusion and examined by light- and electron microscopy and by immunohistochemistry. RESULTS: Tubular degeneration started with cellular hypotrophy in the proximal tubule. Hypotrophy appeared to follow obstruction of the initial proximal tubule by a cellular crescent. Whereas induction of intercellular adhesion molecule-1 (ICAM-1) was diffuse in glomerulonephritic mice, expression of CD44 and vascular cell adhesion molecule-1 (VCAM-1) appeared to be restricted to degenerating tubules. Interstitial inflammation developed in the vicinity of degenerating tubules. Inflammatory infiltration of tubules themselves was observed only in late stages of tubular degeneration. CONCLUSION: In a similar manner as described earlier for focal segmental glomerulosclerosis, in crescentic glomerulonephritis nephron loss can be initiated by the progression of a glomerular lesion into the proximal tubule. Interstitial inflammation might be rather a consequence than the cause of tubular degeneration. PMID- 12631124 TI - Acute renal failure leads to dysregulation of lung salt and water channels. AB - BACKGROUND: Renal ischemia/reperfusion (I/R) injury and the acute respiratory distress syndrome (ARDS) frequently coexist in the intensive care setting, and this combination is associated with a high mortality. Recent experimental data demonstrate that renal I/R injury leads to an increase in pulmonary vascular permeability, similar to that observed in ARDS. However, the effects of renal I/R injury on alveolar fluid clearance-of potential importance in the setting of increased permeability-are unknown. We investigated the effects of renal I/R injury on pulmonary epithelial sodium channel (ENaC), Na,K-ATPase and aquaporin expression as a first step in addressing this question. METHODS: Sprague Dawley rats were subjected to four protocols: (1) surgery for bilateral I/R injury, (2) sham surgery, (3) surgery for unilateral I/R injury, or (4) bilateral nephrectomy. Lung tissue was examined for Na channel, Na,K-ATPase, aquaporin-1, and aquaporin-5 expression. Northern and Western blots were performed. RESULTS: Renal I/R injury and bilateral nephrectomy both led to marked down-regulation of pulmonary ENaC, Na,K-ATPase and aquaporin-5 but not aquaporin-1 compared to sham surgery. These changes were not influenced by the animals' volume status. In contrast, unilateral I/R with an intact contralateral kidney did not lead to down regulation of channel down-regulation. CONCLUSIONS: Ischemic acute renal failure leads to down regulation of pulmonary ENaC, Na,K-ATPase and aquaporin-5, but not aquaporin-1. Since bilateral nephrectomy but not single kidney I/R injury also leads to lung changes, these changes are likely mediated by systemic effects of acute renal failure (ARF), such as "uremic toxins," rather than reperfusion products. These changes may modulate lung dysfunction, susceptibility to lung injury, or both. PMID- 12631125 TI - Low protein diet mediated renoprotection in remnant kidneys: Renal autoregulatory versus hypertrophic mechanisms. AB - BACKGROUND: The mechanism of low protein diet conferred renoprotection in the ablation model remains controversial. Blockade of glomerular hypertrophy, reduced preglomerular vasodilation, and preserved autoregulation have all been postulated. The potential differential impact of calcium channel blockers on these mechanisms and glomerulosclerosis was examined. METHODS: Rats with 5/6 renal ablation received either a 25% standard protein diet, an 8% low protein diet and a low protein diet with either verapamil or amlodipine. Renal autoregulatory and morphometric studies were performed at 3 weeks before the development of significant injury, and the assessment of glomerulosclerosis after 7 weeks of continuous blood pressure radiotelemetry in additional rats. RESULTS: The preserved renal autoregulation in low protein rats was abolished by both calcium channel blockers, with the impairment being either comparable to (low protein + verapamil) or greater than the standard protein rats (low protein + amlodipine). Neither calcium channel blocker blocked the inhibitory effects of low protein diet on renal blood flow, kidney weight, and glomerular volume. Results (mean +/- SE) for glomerular volume (microm-3x 10(-6)): low protein (N = 11), 1.6 +/- 0.1; low protein + verapamil (N = 10), 1.7 +/- 0.1; low protein + amlodipine (N = 12), 1.7 +/- 0.2; versus standard protein (N = 10), 2.2 +/- 0.1; P < 0.05. Only amlodipine, but not verapamil, reduced average systolic blood pressure (143 +/- 2 mm Hg versus low protein rats, 168 +/- 5 mm Hg, and standard rats, 170 +/- 6 mm Hg; P < 0.01). Nevertheless, the glomeruloprotection seen in low protein (N = 15) as compared to standard protein (N = 14) rats (9%+/- 3% versus 28%+/- 6% glomerulosclerosis; P < 0.01) was abolished in both low protein + verapamil (N = 14, 32%+/- 7%) and low protein + amlodipine rats (N = 16, 27%+/- 7%). CONCLUSIONS: Preservation of renal autoregulation and not inhibition of hypertrophy is the critical component in low protein diet-conferred glomeruloprotection. PMID- 12631126 TI - Effects of angiotensin II receptor antagonism on the renal hemodynamic response to cardiovascular stress. AB - BACKGROUND: To elucidate the effect of the angiotensin type 1 (AT1) receptor antagonist (AT1RA) eprosartan (E) on renal hemodynamics in normotensive and borderline hypertensive subjects, we investigated the hormonal and renal hemodynamic responses during cardiopulmonary stress testing. METHODS: In a prospective, double-blind, randomized, placebo-controlled crossover study, the effects of E on renal plasma flow (RPF), renal blood flow (RBF), glomerular filtration rate (GFR), and the concentration of angiotensin II (Ang II) levels were measured with the subjects at rest and during perturbation of cardiopulmonary baroreceptors using lower body negative pressure (LBNP). Ten normotensive male subjects (NT) versus 14 males with mild hypertension (HT), matched for age and body mass index, who were all free of any medication, were randomly assigned to receive placebo or E 600 mg/day PO for seven days (intake phase 1). After a washout period of four weeks the subjects started the intake of the other substance for seven days in a crossover manner (intake phase 2). The measurements were taken on day 7 of both intake phases. RESULTS: During the LBNP test, RPF and RBF were reduced significantly in all subjects; GFR, however, decreased significantly during cardiopulmonary stress testing in the subjects taking the placebo (P < 0.05) and remained unchanged in those under treatment with AT1RA. Ang II levels increased significantly during cardiopulmonary stress test only in the subjects with hypertension who were on placebo, whereas the Ang II levels did not change in normotensive subjects or those treated with the AT1RA. CONCLUSIONS: The data confirm that with cardiovascular stress simulating orthostasis or volume depletion, subjects with AT1RA can maintain their GFR level, suggesting that AT1RA potentially is renoprotective. Additionally, the neurohumoral system is activated after cardiovascular stress in subjects even at an early stage of hypertension. PMID- 12631127 TI - Endothelial dysfunction and inflammation in asymptomatic proteinuria. AB - BACKGROUND: Proteinuria is associated with vascular risk and a systemic increase in vascular permeability. Endothelial dysfunction occurs early in atherosclerosis and modulates vascular permeability. Vascular risk and chronic inflammation are associated. This study investigates whether the increased vascular permeability in proteinuria reflects systemic endothelial dysfunction and chronic inflammation. METHODS: Twenty-one patients with asymptomatic proteinuria (1.29 g/24 h; range 0.18 to 3.17) and 21 matched controls were studied. Microvascular endothelial function was assessed using acetylcholine iontophoresis. Maximum microvascular hyperemia (MMH) was assessed by flux response to local skin heating. Macrovascular endothelial function was assessed by flow-associated dilation (FAD) in the brachial artery using ultrasound. von Willebrand factor (vWF) was measured as a marker of endothelial activation. Low-grade inflammation was assessed by measurement of circulating C-reactive protein (CRP) values using a high sensitivity assay. RESULTS: FAD was impaired in proteinuric subjects (AP) compared to controls [1.8 (0.2 to 5.3) AP vs. 3.8 (1.5 to 6.2) C %; P = 0.014]. There was no significant difference between groups in MMH or in the response to acetylcholine iontophoresis. The AP group had a higher CRP [4.0 (0.5 to 39.0) AP vs. 0.2 (0.1 to 21.3) C mg/L; P < 0.001] and tendency to higher vWF [101.5 (67.0 to 197.0) AP vs. 77.5 (45.0 to 185.0) C IU/dL; P = 0.046] compared to controls. In the AP, but not control, group there was an inverse correlation between CRP and microvascular function as determined by acetylcholine iontophoresis (r = 0.509; P = 0.018). CONCLUSIONS: In AP subjects there is evidence of macrovascular endothelial dysfunction remote from the kidney and of low-grade inflammation that is associated with microvascular endothelial dysfunction. PMID- 12631129 TI - The myogenic response in uremic hypertension. AB - BACKGROUND: The constriction of resistance arteries in response to an increase in transmural pressure, the myogenic response, is thought to be an important determinant of peripheral vascular resistance and therefore of arterial blood pressure. Since raised peripheral resistance is known to occur in uremic hypertension, abnormal myogenic constriction might be responsible. We sought to assess the myogenic response of resistance arteries from the subtotal nephrectomy rat model of uremic hypertension. METHODS: Uremic Wistar-Kyoto (WKYU) rats, and sham-operated normotensive (WKYC) and spontaneously hypertensive (SHRC) controls were studied in parallel. Skeletal muscle arteries were mounted on a pressure myograph and allowed to develop myogenic constriction. The active internal diameter was measured at increasing lumen pressures from 20 to 200 mm Hg. Vascular smooth muscle then was relaxed in a calcium free solution containing nitroprusside, and the passive internal diameter measured at the same pressure steps. The ratio of active to passive diameter at any given pressure was used to assess the myogenic response. RESULTS: Myogenic constriction was not increased in either WKYU or SHRC compared to WKYC at pressures up to 180 mm Hg. CONCLUSIONS: Increased myogenic tone is not the cause of uremic hypertension. PMID- 12631128 TI - N-acetylcysteine ameliorates renal microcirculation: studies in rats. AB - BACKGROUND: N-acetylcysteine (NAC) administration has been shown to ameliorate experimental acute renal failure induced by ischemia-reflow, and was found to prevent radiocontrast nephropathy in high-risk patients. While the protective effect of NAC has been primarily attributed to scavenging oxygen free radicals, improving renal microcirculation also may play a role in the prevention of acute renal failure. METHODS: This study was designed to explore the effect of NAC on renal microcirculation. Blood pressure, total renal blood flow and selective regional cortical and outer medullary blood flow were continuously monitored in anesthetized Sprague Dawley rats with ultrasonic and laser-Doppler probes during the infusion of NAC (60 mg/kg). RESULTS: In control intact rats blood pressure and renal microcirculation were unaffected by NAC. By contrast, following renal vasoconstriction induced by the radiocontrast agent iothalamate meglumine, NAC decreased total, cortical and medullary vascular resistance by 7 to 10% (P < 0.05). NAC also reduced renal vascular resistance by 16% when given during angiotensin II infusion (P < 0.05). Altered renal microcirculation, induced by the cyclooxygenase inhibitor indomethacin, by the nitric oxide synthase inhibitor, Nomeganitro-l-arginine (L-NAME), or with their combination was partially restored by NAC. Nevertheless, NAC administration failed to attenuate renal function and morphology in a rat model of acute renal failure with selective outer medullary hypoxic injury, induced by indomethacin, L-NAME and iothalamate. CONCLUSIONS: NAC ameliorates renal vasoconstriction, an effect that seems to be mediated by mechanisms other than prostaglandins and nitric oxide. The potential renoprotective outcome of NAC and the role of its vasodilating effect on the pre-constricted renal vasculature should be evaluated further. PMID- 12631130 TI - Impact of renal transplantation on survival in end-stage renal disease patients with elevated body mass index. AB - BACKGROUND: Cadaveric renal transplantation is associated with a survival advantage compared with dialysis patients remaining on the renal transplantation waiting list, but this advantage has not been confirmed in obese end-stage renal disease (ESRD) patients. METHODS: Using data from the USRDS, we studied 7521 patients who presented with ESRD from 1 April 1995 to 29 June 1999 and later enrolled on the renal transplantation waiting list with body mass indices (BMI) >or=30 kg/m(2) at the time of presentation to ESRD, and followed until 6 November 2000. Recipients of preemptive renal transplantation or organs other than kidneys were excluded. Cox non-proportional hazards regression models were used to calculate adjusted, time-dependent hazard ratios (HR) for time to death in a given patient during the study period, controlling for renal transplantation, demographics and comorbidities (Form 2728). RESULTS: The incidence of mortality was 3.3 episodes per 100 patient-years (PY) in cadaveric renal transplantation and 1.9/100 PY in living donor renal transplantation compared with 6.6 episodes/100 PY in all patients on the transplant waiting list. In comparison to maintenance dialysis, both recipients of solitary cadaveric kidneys (HR 0.39, 95% CI 0.33 to 0.47), and recipients of living donor kidneys (HR 0.23, 95% CI 0.16 to 0.34) had statistically significant improved survival. A benefit of cadaveric renal transplantation did not apply to patients with BMI >or=41 kg/m(2) (HR 0.47, 95% CI, 0.17 to 1.25, P = 0.13). CONCLUSIONS: Obese patients on the renal transplant waiting list had a significantly lower risk of mortality after renal transplantation compared with those remaining on dialysis. PMID- 12631131 TI - C-reactive protein is associated with renal function abnormalities in a non diabetic population. AB - BACKGROUND: C-reactive protein (CRP) has recently been introduced in cardiovascular medicine as a predictor of myocardial infarction, stroke and peripheral artery disease in different populations. We hypothesized that elevated CRP levels are associated with renal function abnormalities. METHODS: To test this hypothesis, we studied the relationship between CRP levels and renal function loss measured as diminished creatinine clearance in a large non-diabetic population (7317 subjects). In addition, the associations and confounding effects of established renal risk factors that could explain the association between CRP and diminished renal filtration were studied. Also, the association of CRP with early alterations in renal function, such as those evidenced by a relatively high glomerular filtration ("hyperfiltration"), was examined. CRP levels were divided in quartiles. Subjects with CRP levels within the first quartile were defined as the reference group. Diminished renal filtration and hyperfiltration were defined as a creatinine clearance below or exceeding two times the prediction interval of the age- and sex-related reference group. RESULTS: Elevated CRP levels were positively associated with cardiovascular and renal risk factors: age, body mass index, blood pressure, serum cholesterol level, smoking, plasma glucose level and elevated urinary albumin excretion. Elevated CRP was positively associated with diminished filtration (OR 1.8; 95% CI 1.2 to 2.6). In multivariate analyses, CRP was independently associated with a diminished filtration (OR 1.9; 95% CI 1.3 to 2.9). Interestingly, CRP also was associated with hyperfiltration (highest quartile, OR 1.7; 95% CI 1.2 to 2.5). However, body mass index accounted for most of the relationship between CRP and hyperfiltration. CONCLUSIONS: As in cardiovascular disease, CRP appears to be a risk marker for renal function loss. The mechanism of this relationship remains to be clarified. However, the association between CRP, body weight, and a relatively elevated creatinine clearance is a hypothesis-generating finding, suggesting that early inflammatory processes related to high body fat may predispose the kidney to glomerular hyperfiltration-related renal function loss. PMID- 12631132 TI - Bone mineral density and urine calcium excretion among subjects with and without nephrolithiasis. AB - BACKGROUND: Bone mineral density (BMD) is reduced among patients with idiopathic hypercalciuria (IH) and nephrolithiasis. To disentangle effects of diet, stone formation, and physiology upon BMD, we studied vertebral and femoral neck BMD among relatives of hypercalciuric stone formers, and contrasted those with to those without stones. METHODS: Among 59 subjects from 11 families, vertebral and femoral neck BMD, diet calcium intake, urine excretions of calcium, sodium, ammonium, titratable acid, sulfate, urea nitrogen, and serum levels of calcitriol and markers of bone turnover were studied. RESULTS: Stone formers (SF) consumed less calcium than non-stone formers (NSF). Spine and femoral neck BMD z-scores varied inversely with urine calcium loss and urine ammonium excretion among SF but not NSF. No correlations of BMD z-score were found for bone markers, calcitriol, or any of the other measurements. CONCLUSION: SF consumed less calcium, presumably to prevent more stones, and displayed a bone mineral responsiveness to calcium loss and ammonium excretion not present among NSF, who ate more calcium. Lowered calcium consumption in IH, perhaps in response to stone formation, alters bone responses in a direction that can predispose to mineral loss and eventual fracture. PMID- 12631133 TI - Renal survival and prognostic factors in patients with PR3-ANCA associated vasculitis with renal involvement. AB - BACKGROUND: Severe renal disease is a feature of anti-neutrophil cytoplasmic antibodies (ANCA)-associated small-vessel vasculitis. We evaluated patient and renal survival and prognostic factors in patients with PR3-ANCA associated vasculitis with renal involvement at diagnosis during long-term follow-up. METHODS: Eighty-five patients were diagnosed between 1982 and 1996 and followed until 2001 allowing >or=5 years of follow-up. All patients were treated with prednisolone and cyclophosphamide. Univariate and multivariate analyses with patient and renal survival as dependent variables were performed. RESULTS: Of the 85 patients in this study, 17 (20%) died within one year after diagnosis. Of the 25 patients (29%) who were dialysis dependent at diagnosis, two remained dependent and two again became dialysis dependent after less than one year; nine died early without renal recovery. Risk factors for death occurring within one year in univariate analysis (RR, 95% CI) were age>65 years (6.5, 1.6-13.7) and dialysis dependency at diagnosis (3.6, 1.0-13). Twenty patients died beyond one year during the long-term follow-up. Male gender (4.7, 1.6-10) and developing dialysis dependency during follow-up (4.1, 1.4-12) were associated with poor outcome. Risk factor for renal failure within one year was dialysis dependency at diagnosis (29, 3.6-229). Of 64 patients dialysis independent one year after diagnosis, 12 patients became dialysis dependent during follow-up. A renal relapse was strongly associated with development of renal failure in long-term follow-up (17, 3.5-81). CONCLUSIONS: Early death and failure to recover renal function in PR3-ANCA associated vasculitis is associated with age> 65 years and dialysis dependency at diagnosis. Long-term renal survival is determined by renal relapses during follow-up only. Slow, progressive renal failure without relapses is rarely observed in this group. PMID- 12631134 TI - Epidemiological study of kidney survival in autosomal dominant polycystic kidney disease. AB - BACKGROUND: It is unknown whether the substantial increase in research, identification of risk factors for renal progression, greater antihypertensive armamentarium including inhibitors of the renin-angiotensin-aldosterone system (RAAS) and enhanced educational information have impacted the progression of autosomal dominant polycystic kidney disease (ADPKD) renal disease. METHODS: An epidemiological study involving 513 ADPKD subjects was performed. The hypothesis tested was that over two separate periods, 1985 to 1992 versus 1992 to 2001, a significant slowing of renal function loss in ADPKD patients would be demonstrated in association with improved blood pressure (BP) control and inhibition of the RAAS as instituted by their primary care physicians. RESULTS: ADPKD males and females in the later cohort (1992 to 2001) had longer mean and median survival times to ESRD than males and females in the earlier cohort (1985 to 1992). Analysis revealed that both males and females in the later cohort had significantly lower diastolic blood pressure (DBP) and mean arterial pressure (MAP) values than males and females in the earlier cohort. ADPKD male and female patients in the later cohort used significantly more angiotensin converting enzyme inhibitors (ACEIs) than ADPKD male and female patients in the earlier cohort. CONCLUSIONS: These results demonstrate a significant slowing of ADPKD renal progression in both male and female patients that was associated with a significantly lower MAP and increased use of ACEIs in the later cohort (1992 to 2001) as compared to the early cohort (1985-1992). PMID- 12631135 TI - Apolipoprotein E in idiopathic nephrotic syndrome and focal segmental glomerulosclerosis. AB - BACKGROUND: Hyperlipemia characterizes nephrotic syndrome (NS) and contributes to the progression of the underlying nephropathy. The data in the literature support an implication of apolipoprotein E (apoE) in both hyperlipemia and focal segmental glomerulosclerosis (FSGS), a malignant condition associated with NS. METHODS: The apoE genotype was determined in 209 nephrotic patients, who were classified according to age and their response to steroids as resistant children (N = 96) and adults (43), and steroid dependent (33) and steroid responder (37) children. A total of 123 presented the histological features of FSGS. In a subgroup of 28 patients, serum and urinary levels of apoE and renal deposits were evaluated by immunofluorescence. RESULTS: The allelic frequencies of the three major haplotypes epsilon2, epsilon3, and epsilon4 were the same in nephrotic patients versus controls, and homozygosity for epsilon3epsilon3 was comparably the most frequent genotype (70 vs. 71%) followed by epsilon3epsilon4, epsilon2epsilon3, epsilon2epsilon4, epsilon4epsilon4. Serum levels of apoE were fivefold higher in NS and in FSGS patients than in controls, with a direct correlation with hypercholesterolemia and proteinuria. ApoE genotypes did not influence serum levels. Urinary levels were 1/10,000 of serum with an increment in nephrotic urines. Finally, immunofluorescence demonstrated the absence of apoE in sclerotic glomeruli, while comparably nephrotic patients with membranous nephropathy had an increased glomerular expression of apoE. CONCLUSIONS: ApoE is dysregulated in NS with a marked increment in serum, which is a part of the complex lipid metabolism. Down-regulation of glomerular apoE instead is a peculiarity of FSGS and may contribute to the pathogenesis of the disease. The normal distribution of apoE genotypes in nephrotic patients with FSGS excludes a pathogenetic role of genetic variants. PMID- 12631136 TI - Grade of chronic renal failure, and acute and long-term outcome after percutaneous coronary interventions. AB - BACKGROUND: Patients with moderate chronic renal failure have recently been identified to suffer from a markedly higher mortality after percutaneous coronary intervention (PCI). We focused on the outcome of PCI patients with just mildly elevated creatinine levels of 1.1 to 1.5 mg/dL. METHODS: Data of all PCI patients of the years 1998 to 1999 were analyzed. Follow-up was performed by a questionnaire sent to all patients. RESULTS: During this period, PCI was performed in 1049 patients. Long-term follow-up (1184 +/- 10 days) was 99.6% complete. Total mortality increased continuously by each creatinine increment of 0.1 mg/dL above 1.0 mg/dL, with a significant difference at 1.3 mg/dL compared to patients with or=5 years after a biopsy. RESULTS: A total of 102 de novo cancers were found in 1958 patients. These cancers represent a two- to threefold excess of the expected number at <1 and 1 to 4, but not >or=5 years after a biopsy. Non-Hodgkin's lymphomas were observed six to eight times more than expected. Cancer excess was seen in glomerulonephritides with a known or suspected virus etiology. CONCLUSIONS: The excess cancer rate could be the result of underlying undiagnosed tumors whose antigens have initiated glomerulonephritis, or the immunosuppressive therapy that initiated or energized tumor cells. Based on the findings in our study, there is some support for an association to persistent viruses causing first the glomerulonephritides and then the malignancies, perhaps through a common pathogenesis. This calls for other studies to be done that are specifically designed to investigate this issue, with more data on patient characteristics and confounders. PMID- 12631140 TI - High glucose levels inhibit focal adhesion kinase-mediated wound healing of rat peritoneal mesothelial cells. AB - BACKGROUND: The peritoneum is progressively denuded of its mesothelial cell monolayer in patients on continuous ambulatory peritoneal dialysis (CAPD). These alterations of the mesothelium cause membrane dysfunction and progressive peritoneal fibrosis. Integrins regulate cell motility and play an important role in wound healing. We investigated the effects of high glucose on the regeneration process of the peritoneal mesothelial cell monolayer using cultured rat peritoneal mesothelial cells (RPMC). METHODS: The effects of glucose or mannitol on the regeneration of RPMC and formation of focal adhesions were examined by in vitro wound healing assay and immunocytochemistry, respectively. Activities of focal adhesion kinase (FAK) and its downstream p130Cas were examined by Western blotting. Effects of wild-type and dominant-negative FAK on RPMC migration were examined by a transient transfection assay. RESULTS: Cell migration over fibronectin (FN) was clearly inhibited in culture media containing high glucose (28 to 140 mmol/L). RPMC formed focal adhesions on FN in the presence of a regular glucose concentration (5.6 mmol/L); however, tyrosine phosphorylation of FAK and p130Cas and formation of focal adhesions observed by FAK and vinculin staining were substantially inhibited by high glucose. Mannitol also induced significant inhibitory effects, but these were milder than those of glucose. Transfection of dominant-negative FAK inhibited cell migration in a regular glucose concentration, whereas overexpression of wild-type FAK abrogated glucose induced inhibition of cell migration. CONCLUSIONS: Our results demonstrate that high glucose concentrations as well as high osmolarity inhibit FAK-mediated migration of mesothelial cells, and suggest that dialysates containing high glucose concentrations may cause peritoneal damage by inhibiting wound healing of the mesothelial cell monolayer. PMID- 12631141 TI - Effects of uremic ultrafiltrate on the regulation of the parathyroid cell cycle by calcitriol. AB - BACKGROUND: Calcitriol (CTR) is used in the treatment of hyperparathyroidism secondary to renal failure because it decreases parathyroid hormone (PTH) synthesis and parathyroid cell proliferation. Previous studies in tissues other than parathyroids have demonstrated that uremic factors affect the action of CTR on the target cells. We questioned whether the uremic milieu interferes with the inhibition of parathyroid cell proliferation by CTR. METHODS: Studies were performed in vitro using freshly excised normal dog parathyroid tissue incubated for 24 hours with and without CTR and in the presence of either total uremic ultrafiltrate (UUF) from uremic patients or high-pressure liquid chromatography (HPLC)-derived fractions (hydrophilic compounds eluting early and hydrophobic compounds eluting late) of this UUF (F1 to F4). Parathyroid cell proliferation was assessed by flow cytometry. RESULTS: The addition of CTR 10-8 and 10-7 mol/L to parathyroid tissue produced an inhibition of the proliferation that was prevented in the presence of UUF. In a medium containing CTR 10-8 mol/L, the addition of F1, F2 and F3, but not F4, prevented the CTR-induced inhibition of parathyroid cell proliferation. With CTR 10-7 mol/L, the inhibition of proliferation was observed even in the presence of F1, F2 and also F4, but was prevented by F3. Uric acid (7 mg/dL), indoxyl sulfate (5 mg/dL) and p-cresol (1.4 mg/dL), which coeluted with F1, F2 and F4, respectively, did not interfere with the inhibitory action of CTR 10-7 mol/L; however, the addition of phenol (0.14 mg/dL), which coeluted with F3, prevented the CTR-induced inhibition of parathyroid cell proliferation. CONCLUSIONS: The presence of uremic toxins prevents the inhibition of parathyroid cell proliferation induced by calcitriol. PMID- 12631142 TI - Influenza vaccine delivery and effectiveness in end-stage renal disease. AB - BACKGROUND: Influenza vaccination rates in the general population have been associated with improved outcomes, yet high-risk populations, such as end-stage renal disease (ESRD) patients, have received little attention in determining the potential benefits. This report assessed the frequency and effectiveness of influenza vaccination, while also assessing disparities in vaccination rates in the ESRD population. METHODS: Using the United States Renal Data System research files containing claims for all Medicare ESRD patients, vaccination rates and outcomes among vaccinated and unvaccinated persons for the 1997 to 1998 and 1998 to 1999 influenza seasons were compared after adjustment for baseline demographic factors and health characteristics. RESULTS: Vaccination rates in the ESRD population were less than 50% for each season. Influenza vaccination rates were lower in non-whites, women, younger patients, and peritoneal dialysis patients. Influenza vaccination was associated with a lower risk for hospitalization and death. CONCLUSIONS: Despite universal coverage of free influenza vaccination, the ESRD population had a less than 50% vaccination rate for the years 1997 to 1998 and 1998 to 1999 as demonstrated by Medicare billing data. Substantial differences were found in vaccination rates among non-whites and peritoneal dialysis patients. This study confirms that the ESRD populations benefit from influenza vaccination, suggesting that dialysis providers should take advantage of all opportunities to immunize this high-risk group. PMID- 12631143 TI - A prospective cohort study of incident maintenance dialysis in children: an NAPRTC study. AB - BACKGROUND: Prior studies of dialysis practices and outcomes have included children with varied duration of end-stage renal disease (ESRD). This study evaluated dialysis characteristics, complications, practices, and outcomes in an incident pediatric cohort. METHODS: The cohort was limited to 1992 subjects enrolled in the North American Pediatric Renal Transplant Cooperative Study registry, starting hemodialysis (HD) or peritoneal dialysis (PD) between 1992 and 1998, without prior dialysis or transplantation. RESULTS: At dialysis initiation, the median glomerular filtration rate (GFR; Schwartz formula) was 6 to 11 mL/min/1.73 m2, and 90th percentile was 14 to 25 mL/min/1.73 m2. GFR was not associated with age or race. PD was used in 97% of infants, 70 to 80% of children and 59% of adolescents. Blacks were significantly less likely to be started on PD than whites. Twenty percent of patients switched dialysis modality, largely due to infection, inadequate access or family choice. Younger children received HD almost exclusively through percutaneous catheters, while 57% of children more than six years old were dialyzed with fistula or graft after six months on HD. The prevalence of anemia (Hct <33%) still exceeded 40% after six months of dialysis. The median interval to transplantation was 1.4 years, and was significantly greater in non-white, young, and female patients. Mortality rates (deaths/1000 patient-years) varied with age, from 13.6 in infants to 2.2 in adolescents. CONCLUSION: These data demonstrate considerable variability in patient management across pediatric centers. Prospective studies are needed to determine the optimum adequacy of care among children on dialysis and to identify populations at risk. PMID- 12631145 TI - Congential obstructive nephropathy gets complicated. PMID- 12631144 TI - Effects of carboxy-terminal modifications of proteinase 3 (PR3) on the recognition by PR3-ANCA. AB - BACKGROUND: Autoantibodies directed against neutrophil proteinase 3 (PR3-ANCA) from patients with Wegener's granulomatosis and microscopic polyangiitis recognize conformational epitopes of PR3. During maturation of neutrophils, PR3 undergoes amino-terminal and carboxy-terminal processing. In contrast to amino terminal processing, the effects of carboxy-terminal processing on recognition of PR3 by PR3-ANCA remain unknown. Carboxy-terminally modified or tagged recombinant PR3 (rPR3) molecules may be useful for the refinement of diagnostic assays and for the study of biological processes. METHODS: This study was designed to determine whether 293 cells can be used to express specifically designed carboxy terminal variants of rPR3, and to evaluate the effects of different carboxy terminal modifications on the recognition by PR3-ANCA in the capture ELISA. RESULTS: The rPR3-variants secreted into the media supernatants of transfected 293 cells escaped proteolytic processing. Furthermore, in contrast to the effects of amino-terminal pro-peptide deletion on PR3-ANCA binding, carboxy-terminal modifications (deletion and additions) did not significantly affect recognition by PR3-ANCA. CONCLUSIONS: This expression system is ideally suited for the expression of custom-designed carboxy-terminal rPR3 variants, and major conformational effects of carboxy-terminal modifications seem unlikely. PMID- 12631146 TI - Renoprotection: clues from knockout models of rare diseases. PMID- 12631147 TI - Nuclear factors, hs1,2 enhancer and IgA nephropathy. PMID- 12631148 TI - Venous central catheter and risk of death among hemodialysis patients. PMID- 12631149 TI - Dual blockade of renin-angiotensin system. PMID- 12631151 TI - The antiproteinuric effect of high-dose ramipril: still an open question. PMID- 12631153 TI - The statement that folate supraphysiological levels in uremic patients do not cause harm should not go unchallenged. PMID- 12631155 TI - Tubular injury in glomerular disease. PMID- 12631156 TI - The frequency range of TMJ sounds. AB - There are conflicting opinions about the frequency range of temporomandibular joint (TMJ) sounds. Some authors claim that the upper limit is about 650 Hz. The aim was to test the hypothesis that TMJ sounds may contain frequencies well above 650 Hz but that significant amounts of their energy are lost if the vibrations are recorded using contact sensors and/or travel far through the head tissues. Time-frequency distributions of 172 TMJ clickings (three subjects) were compared between recordings with one microphone in the ear canal and a skin contact transducer above the clicking joint and between recordings from two microphones, one in each ear canal. The energy peaks of the clickings recorded with a microphone in the ear canal on the clicking side were often well above 650 Hz and always in a significantly higher area (range 117-1922 Hz, P < 0.05 or lower) than in recordings obtained with contact sensors (range 47-375 Hz) or in microphone recordings from the opposite ear canal (range 141-703 Hz). Future studies are required to establish normative frequency range values of TMJ sounds but need methods also capable of recording the high frequency vibrations. PMID- 12631157 TI - Clinical quality of removable dentures provided by dentists, denturists and laboratory technicians. AB - The aim of this study was to evaluate the clinical quality of removable dentures of elderly Finnish men, which had been prepared either by dentists, denturists or dental laboratory technicians. The participants comprised 242 denture-wearing subjects, with 231 maxillary and 177 mandibular removable dentures which had been prepared either by dentists, denturists or dental laboratory technicians. Clinical examinations were carried out without the examining dentist knowing who had provided the dentures. Complete dentures which had been illegally provided by laboratory technicians had significantly poorer retention and fitted less well in tuber and alveolar areas than those provided by either dentists or denturists. Complete maxillary dentures which had been provided illegally by laboratory technicians had significantly (P < 0.01) higher occurrence (90%) of some unacceptable characteristics than those (43%) provided by dentists or denturists. The difference between complete mandibular dentures was also obvious, 86% versus 59%, although statistically non-significant. Of those partial maxillary dentures provided by dentists 53% had some unacceptable characteristics, compared with 80% of those illegally provided by denturists or laboratory technicians (NS). In the case of partial mandibular dentures, 36% of those provided by dentists and 32% of those by denturists or laboratory technicians had some unacceptable characteristic (NS). Illegal provision of removable dentures seemed to be related to decreased clinical quality. PMID- 12631158 TI - The effect of three different periodontal pre-treatment procedures on the success of telescopic removable partial dentures. AB - In this prospective study, 120 teeth consisting of maxillary and mandibular canines and premolars were divided into three groups each containing 40 teeth. The teeth were assigned randomly in quadrants to three different periodontal treatment protocols. The first group was treated with professional prophylaxis only. The second group received additional deep scaling. With the third group, additional surgical periodontal flap surgery and scaling was performed. Both papillary bleeding index (PBI) and probing depth (PD) were evaluated before, during and after treatment. During the subsequent prosthetic treatment phase all teeth were then used as telescope abutments supporting a removable prosthesis. The documentation of the attachment level (AL) was then used as a clinical parameter. One year after the incorporation of telescopic removable partial dentures (RPDs), PD, PBI and AL were again evaluated. The resulting periodontal parameters were compared between the different groups using the general linear model (GLM) repeated measures and the Kruskal-Wallis test for non-parametric variables. Differences within the three treatment groups were determined using the t-test, e.g. the Wilcoxon test for dependent variables (P < 0.05). A significant decrease in inflammatory indices (PBI) was found for all types of periodontal treatment (P < 0.03 for all groups). Additionally, the reduction in PD was significant for all of the three groups (P < 0.001 for all groups). The greatest reduction in PD was observed in the group in which a surgical approach was used. Evaluation of the attachment level after the incorporation of the telescopic RPDs showed that tooth position did not influence the periodontal prognosis and that the use of telescopic RPDs exerted no ascertainable negative influence on the periodontium of the abutment teeth. PMID- 12631159 TI - Accuracy of adaptation of thermoformed poly(methyl methacrylate). AB - Thermoformed poly(methyl methacrylate) (PMMA) sheet is used to produce a number of different dental appliances such as stents, occlusal splints and baseplates for occlusal rims. The purpose of the present study was to measure the accuracy of adaptation of Perspex PMMA sheet and to determine the effect of annealing on the accuracy of the thermoformed specimens. The results of the study showed that PMMA can produce specimens that are accurately adapted to the cast. Immersion in water resulted in an increase in the space between the cast and the specimen for both thermoformed and thermoformed and annealed acrylic resin. Annealing of the thermoformed specimens had significantly less increase in space between the cast and the specimens when immersed in water over a period of 3 months. PMID- 12631160 TI - Periodontitis, caries and salivary factors in Sjogren's syndrome patients compared to sex- and age-matched controls. AB - The aims of this investigation were: (i) to study a group of dry mouth Sjogren's syndrome (SS) patients comprising individuals with pathological and non pathological amounts of rest saliva and (ii) to compare these two categories of SS patients with a sex- and age-matched control group with respect to their periodontal and dental status. Thirty-three dry mouth patients and 33 sex- and age-matched patients, referred to the same private dental clinic in southern Norway, were examined for rest and stimulated saliva, as well as their dental and periodontal status. All patients were referred to the local hospital for blood and urine examinations. The dry mouth SS patients were all of the secondary type. Volumes of rest and stimulated saliva were significantly lower in the low saliva SS group compared with the high saliva SS group; the values for immunoglobulin G (IgG) and IgA were similarly lower for the low saliva group, but not for IgM which was significantly higher. The two SS subgroups and their controls were compared for the volume of rest and stimulated saliva, which showed a statistically significant lower volumes for the low saliva SS group compared with the control group. None of the subgroups and their controls differed concerning filled or missing teeth, but the total SS group revealed significantly higher number of missing teeth. The periodontal and dental status did not show any statistically significant differences except for a few scattered higher periodontal level losses of attachment in the SS subgroups. The blood and urine analyses showed statistically significant higher values for sedimentation rate, white blood cell count and haemoglobin in the SS low saliva group compared with the control group while anti-streptolysin was lower. In the high saliva SS group only sedimentation rate and white cell count were higher compared with the control. The conclusions is SS patients do not have an increased risk for developing periodontitis. PMID- 12631161 TI - Condylar movement analysis in subjects with clinically normal temporomandibular joints, utilizing an amorphous sensor. AB - The purpose of this study was to determine the range of condylar movements in normal subjects, by the use of an amorphous sensor. The pair-matched sample consisted of 17 Caucasian males (aged 25.8 +/- 2.5 years) and 17 Caucasian females (aged 25.5 +/- 2.8 years) who had no subjective or objective symptoms related to temporomandibular joint (TMJ) sounds. Bilateral condylar and jaw movements were recorded simultaneously. The results showed that the typical condylar movement points plotted on the X-Y coordinates indicated a uniform or approximately straight line, that is X = Y. The mean +/- standard deviation (s.d.) for the maximum velocity of condylar movement during the opening and closing phases was 32.6 +/- 16.9 and 39.8 +/- 21.5 mm s-1, respectively. The mean +/- s.d. for the degree of jaw opening at the turning point of condylar movement was 36.9 +/- 21.7%. Significant correlations existed between any two measurements of maximum velocity of condylar movement. Moreover, significant correlations existed between the left and right sides in the degree of jaw opening at the turning point of condylar movement. These results suggest that a range of normal values of condylar movements can be developed when utilizing the amorphous sensor method. PMID- 12631163 TI - Mandibular movement and frontal craniofacial morphology in orthognathic surgery patients with mandibular deviation and protrusion. AB - The present study was conducted to investigate the relationship between mandibular movement (lateral excursion and masticatory movements) and craniofacial morphology in 16 patients with mandibular deviation, using a six degrees-of-freedom measuring device. (i) Mandibular deviation was found to be significantly related to frontal maxillary and occlusal plane angles. (ii) Three dimensional non-working condylar and incisal path lengths were longer during the lateral excursion to the non-deviated side than to the deviated side, and the incisal path moved antero-inferior. (iii) The lateral motion range of the incisal path was wider during masticatory movement on the non-deviated side than on the deviated side, and the molar and non-working condylar path lengths corresponding to the lateral range of the incisal path were also longer on the non-deviated side. The group with posterior crossbite showed a significantly smaller horizontal range of incisal path, and also significantly smaller frontal projected incisal and molar path angles during masticatory movement on the deviated side than on the non-deviated side. These results suggest that lateral excursion and masticatory movements could be related to craniofacial morphology and posterior crossbite. PMID- 12631162 TI - Comorbidity of internal derangement of the temporomandibular joint and silent dysfunction of the cervical spine. AB - The aim of this evaluation was to examine correlations between internal derangement of the temporomandibular joint (TMJ) and cervical spine disorder (CSD). A prospective controlled clinical study was carried out. Thirty patients with signs and symptoms of internal derangement but without any subjective neck problems and 30 age- and gender-matched control subjects without signs and symptoms of internal derangement were examined. The investigation of the temporomandibular system was carried out using a 'Craniomandibular Index'. Afterwards an examiner-blinded manual medical investigation of the craniocervical system was performed. This included muscle palpation of the cervical spine and shoulder girdle as well as passive movement tests of the cervical spine, to detect restrictions in the range of movement as well as segmental intervertebral dysfunction. The internal derangement of the TMJ was significantly associated with 'silent' CSD (t-test, P < 0.05). Patients with raised muscle tenderness of the temporomandibular system exhibited significantly more often pain on pressure of the neck muscles than patients without muscle tenderness of the temporomandibular system (t-test, P < 0.05). As a result of the present study, for patients with internal derangement of the TMJ an additional examination of the craniocervical system should be recommended. PMID- 12631164 TI - Validation of the clinical diagnostic criteria for temporomandibular disorders for the diagnostic subgroup of degenerative joint disease. AB - Research is needed to assess the validity of the Clinical Diagnostic Criteria for Temporomandibular Disorders (CDC/TMD). The purpose of this study was to test the reliability of the clinical diagnosis of temporomandibular joint (TMJ) degenerative joint disease (DJD) as compared with the magnetic resonance imaging (MRI) 'gold standard'. The TMJ DJD group comprised 48 joints in 24 consecutive patients who were assigned a clinical bilateral diagnosis of TMJ DJD. The TMJ non DJD group consisted of 82 joints in 41 consecutive patients without a TMJ-related diagnosis of TMD. Bilateral sagittal and coronal MR images were obtained subsequently to establish the corresponding diagnosis of degenerative joint changes. An MRI diagnosis of osteoarthrosis (OA) was defined by the presence of flattening, subchondral sclerosis, surface irregularities, and erosion of the condyle or presence of condylar deformities associated with flattening, subchondral sclerosis, surface irregularities, erosion and osteophyte. For the CDC/TMD interpretations, the positive predictive of DJD for OA was 67%, and for the presence of degenerative joint changes 88%. The overall diagnostic agreement for DJD was 44.6% with a corresponding K-value of 0.01. Most of the disagreement was due to false-negative interpretations of asymptomatic joints. The results suggest CDC/TMD to be predictive for degenerative joint changes but insufficient for determination of OA. Patients assigned a clinical TMJ-related diagnosis of DJD may need to be supplemented by evidence from MRI to determine the presence or absence of OA. PMID- 12631165 TI - Influence of abrasion in clinical manifestation of human dental erosion. AB - The influence of abrasion from oral soft tissues on softened enamel lesion remineralization and erosion development was investigated. Using orange juice, softened enamel lesions were produced on 20 human premolars assigned randomly to 10 volunteers. Sections used as control and two test slabs were cut from each tooth. One of the two slabs from each tooth had an appliance built on it, which protected the lesion from abrasion. The two slabs (with/without appliance) were bonded to the palatal surfaces of upper right and left lateral incisor teeth of the participants who chewed sugar-free gum four times daily. After 28-day intra oral exposure, mineral loss (Delta Z) and lesion depth (ld) in both control and test samples were quantified using transverse microradiography, and the data was analysed by paired t-test. Delta Z was significantly lower in lesions with appliance (protected), but higher in lesions without appliance (unprotected) when compared with control (unexposed). Similar pattern was observed with lesion depth. In unprotected slabs the lesions were abraded resulting in eroded enamel lesions. It was concluded that erosion observed clinically is the combined effect of demineralization of the tooth surface by an erosive agent and abrasion of the demineralized surface by surrounding oral soft tissues and through food mastication. Abrasion from oral soft tissues can contribute to site-specificity of dental erosion. PMID- 12631166 TI - Effectiveness of an oral health promotion programme at the workplace. AB - The purpose of the study was to evaluate the effectiveness of an oral health promotion programme at the workplace. The programme was given once a year at offices or factories, which was voluntary and free for all employees. The programme consisted of clinical examinations followed by oral health guidance, oral hygiene instruction and oral prophylaxis of anterior lower teeth. Oral health status was compared by the times of participation in the programme. It was shown that three times or more participants in the programme had fewer decayed, missing and filled teeth (DMFT) and lower percentage of Community Periodontal Index (CPI) sextants 3 and 4. The oral health promotion programme was effective in keeping or maintaining good oral health among workers. In addition to current activities, the programme should include education to motivate subjects to receive regular check-ups. PMID- 12631167 TI - Mechanical properties of calcium phosphate based dental filling and regeneration materials. AB - The objective of this study was to compare the mechanical properties of calcium phosphate cements (CPC) for possible dental applications with varied liquid and powder compositions under the same testing condition. Cements studied in this experiment were divided into two groups of CPC not containing polymer and polymeric CPC (PCPC). Cement powder was formed by combining equimolar amounts of dicalcium phosphate anhydrous and tetracalcium phosphate, or acrylic resin polymer powder mixture. The CPC specimens for the compressive strength (CS) and diametral tensile strength (DTS) measurements were prepared by mixing powder and liquid for 30 s with a powder/liquid ratio of 3:1, and subsequently packing the paste into a brass mould. The specimens were kept at 37 degrees C and 100% relative humidity for 24 h before measurements were conducted on a Universal Testing Machine with a cross-head speed of 1 mm min-1. The CS of CPC was 0.14 10.29 MPa and that of PCPC was 0.26-117.58 MPa. The DTS of CPC was 0.10-4.56 MPa and that of PCPC was 0.07-22.54 MPa. The CS and DTS were very diverse depending on the composition of powder and liquid. Some compositions showed higher values than commercial liners. Thus compositions of 2% carboxymethyl cellulose + 35% citric acid in phosphate buffered saline (PBS), 20% gelatin in PBS, 2% sodium alginate in PBS, 20-40% aqueous acrylic-maleic copolymer solution, and some of the HPMC and PMVE-Ma solutions exhibited promising formulae for dentine regenerating materials. Acrylic resin-PCPC group showed generally higher CS and DTS values. Based on this study, further studies on the reaction with odontoblast and resultant dentine regeneration should be performed using promising compositions. PMID- 12631168 TI - A comparison of the in vitro cytotoxicity of two root canal sealers. AB - The purpose of this study was to compare the cytotoxicity of an epoxy resin-based sealer (AH Plus) and a silicone-based sealer (Roeko Seal Automix, RSA). Cytotoxicity was assessed using the MTT assay for mitochondrial enzyme activity and haemocytometer viable cell counting after 24, 48 and 72-h exposure to L929 cells. AH Plus and RSA showed no statistically significant difference in the degree of toxicity. Both sealers had a low toxic influence on the cells during the experimental period. This study indicates that epoxy resin-based sealer AH Plus and the silicone-based sealer RSA have similar levels of cytotoxicity to mouse fibroblasts. PMID- 12631169 TI - Effect of scanning level and muscle condition on ultrasonographic cross-sectional measurements of the anterior masseter muscle. AB - With the disadvantage of computed tomography showing cumulative biological effects and magnetic resonance imaging posing a problem in clinical availability and cost, several authors described the technique of ultrasonography to measure non-invasively local cross-sectional dimensions (LCSDs) of masseter muscle sites. However only few studies addressed the issue of 'technique-related factors for intra- and inter-observer reliability' to gain more consistent testing and diagnosis. The purpose of the present study was to determine (1) whether the scanning level and/or the muscle condition may affect LCSD measurements and (2) whether measurements made at identical levels may be reproducible. The study included 35 subjects with signs and symptoms of temporomandibular disorders. Bilateral ultrasonographic investigation was performed with a linear (B-scan) 7.5 Mhz small-part transducer to register LCSDs of the anterior masseter muscle on three different levels. Scans were made on the relaxed and contracted muscle. Measurements were made in two sessions with a time interval of at least 5 min. Data were analysed for reproducibility by using the intra-class correlation coefficient (ICC) and the method error (ME). Scanning level and muscle condition had a significant effect on muscle measurements (P = 0.000). There was no difference in LCSD between the right and left muscle (P = 0.531). Measurements recorded at a given site were consistent across the testing sessions (P = 0.058). The scanning level with highest reproducibility was halfway between the origin and insertion (ICC = 0.92; ME = 6.2%). The data suggest that ultrasonography is a reliable method for measuring LSCDs of the anterior masseter muscle. PMID- 12631170 TI - The pH changes of four different calcium hydroxide mixtures used for intracanal medication. AB - The purpose of this study was to determine the pH values of calcium hydroxide mixtures in combination with either distilled water, two different local anaesthetic solutions or physiological saline solution, which are commonly used as a dressing materials during endodontic treatment. The pH of each vehicle prior to mix with calcium hydroxide and subsequently each prepared combination was determined using a digital pH meter. The pH measurements were determined at 3 min, 10 min, 1 h, 24 h, 48 h and 7 days. The results of this study indicated that four different water-based vehicles demonstrated similar pH changes range within pH 11-12 when mixed with calcium hydroxide. PMID- 12631171 TI - The periodontal status of pregnant women and its relationship with socio demographic and clinical variables. AB - The aims of this study were to assess the periodontal health and oral hygiene status of pregnant women and non-pregnant controls and to evaluate the effect of socio-demographic and other variables on such periodontal status. A total of 400 women, of whom 200 were pregnant (mean age=30 years, s.e.m.=0.05) and 200 non pregnant controls (mean age=32 years, s.e.m.=0.05) were chosen at random from four health centres. The clinical parameters used were the Silness and Loe plaque index (Pl.I), Loe and Silness gingival index (GI), probing pocket depth (PPD), probing attachment level (PAL), any relationship to socio-demographic (age, level of education and professional level) and clinical variables (gestation period, previous pregnancy and vomiting during pregnancy) was evaluated. The results showed that pregnant women had significantly higher GI and PPD scores (P < 0.005) but with no statistically significant differences in PAL or Pl.I (P > 0.01) compared with non-pregnant controls. Increased age, lower level of education and non-employment were associated with significantly higher GI and PPD scores (P < 0.01). All these clinical parameters increased in parallel with the increase in the stage of pregnancy, reaching their maximum at the eighth month. Women with previous or multiple pregnancy had statistically significantly higher GI and PPD scores than those who were pregnant for the first time (P < 0.01), but with no statistically significant differences in Pl.I or PAL scores (P > 0.05). Also, women who vomited during pregnancy had significantly higher GI and PPD scores compared with those who did not vomit (P < 0.05). It is concluded that gingival inflammatory symptoms are aggravated during pregnancy and are related to increased age, lower level of education and non-employment. Therefore periodontal preventive programmes are very important for pregnant women. PMID- 12631173 TI - A placebo-controlled multi-centred evaluation of an anaesthetic gel (Oraqix) for periodontal therapy. AB - AIMS: Six Canadian dental schools investigated the ability of a thermosetting gel containing 25 mg/g prilocaine and 25 mg/g lidocaine as active agents to produce analgesia in periodontal pockets utilizing a randomized, double-blind, placebo controlled study. MATERIALS AND METHODS: The study consisted of 130 patients, each of whom received the active or placebo gel in periodontal pockets in one quadrant of the mouth for 30 s prior to periodontal debridement (scaling and root planing). Pain was measured using both a 100-mm Visual Analogue Scale (VAS) and a Verbal Rating Scale (VRS). RESULTS: The median VAS pain score for the patients treated with the anaesthetic gel was 5 mm (range 0-85 mm) as opposed to 13 mm (range 0-79 mm) in the placebo-treated patients (P=0.015). There was no significant difference in the percentage of patients reporting no or mild pain (78% and 76% for the anaesthetic gel and placebo, respectively). No significant differences were seen in patient demographics, or mandible versus maxilla. CONCLUSIONS: The VAS pain scores showed that the anaesthetic gel 5% was statistically more effective than the placebo in reducing pain during periodontal debridement. PMID- 12631174 TI - Osteogenesis by guided tissue regeneration and demineralized bone matrix. AB - AIM: To evaluate in a discriminating capsule model whether bone formation by guided tissue regeneration (GTR) may be influenced by concomitant implantation of demineralized bone matrix (DBM). MATERIALS AND METHODS: Thirty 4-month-old male albino rats of the Wistar strain were used in the study. Following surgical exposure of the mandibular ramus, a hemispherical, Teflon capsule (5.0 mm in diameter), loosely packed with a standardized amount of DBM, was placed with its open part facing the lateral bone surface of the ramus. At the contralateral side, an empty capsule was placed, serving as control. After healing periods of 15, 30, and 120 days, groups of 10 animals were sacrificed and 40-70 microm thick undecalcified sections of the capsules were produced. In the sections, the cross sectional areas of (1) the space created by the capsule, (2) newly formed bone, (3) DBM particles, (4) loose connective tissue as well as the (5) height of the capsules, and (6) that of the newly formed bone were measured. RESULTS: Increasing bone fill was observed in both test and control sites from 30 to 120 days. After 30 days of healing, the mean amount of bone was approx. 3% of the cross-sectional area of the capsules at the test sites while it was 8% in the control sites (p<0.05). However, no statistically significant differences were observed between the test (46%) and control (64%) sites after 120 days regarding any of the measured parameters (p>0.05). The newly formed bone in the DBM group at 120 days, on the other hand, appeared more dense than that in the control capsules. CONCLUSION: DBM used as an adjunct to GTR did not provide any added effect on bone formation but increased the density of the newly formed bone. PMID- 12631175 TI - HHV-6, HHV-7, HHV-8 in gingival biopsies from chronic adult periodontitis patients. A case-control study. AB - BACKGROUND: Recent reports have suggested that various herpesviruses may be involved in the occurrence and progression of different forms of periodontal disease. OBJECTIVE: The objective of the present study was to investigate the presence of the novel herpesviruses HHV-6, HHV-7 and HHV-8 in gingival biopsies from patients affected by chronic adult periodontitis. As control, gingival biopsies from periodontally healthy subjects were analysed. MATERIALS AND METHODS: Gingival biopsies were harvested from 23 volunteers: 13 patients affected by chronic adult periodontitis (CAP) and 10 periodontally healthy subjects. Each CAP patient contributed two biopsies involving the epithelium and connective tissue facing the sulcus/periodontal pockets: one biopsy from a site having a probing pocket depth (PPD) > or =5 mm and presenting with bleeding upon probing (affected site) at the time of biopsy collection, and the other biopsy from a site with PPD< or =3 mm and without bleeding on probing (nonaffected site). After DNA extraction, nested PCR was used in herpesvirus identification. RESULTS: HHV-6 DNA sequences were detected in one non-affected site (8%) and no affected sites (0%) of CAP patients. One biopsy (10%) in healthy subjects revealed HHV-6 positivity. Tissue specimens in 10/13 CAP patients (77%) and 7/10 healthy subjects (70%) contained HHV-7 DNA. HHV-7 prevalence in affected and nonaffected sites of CAP patients was 77% and 54%, respectively. HHV-8 was detected in 7.7% of CAP patients and 0% of healthy subjects. CONCLUSIONS: Gingival tissue may act as a reservoir for HHV-7. A high prevalence of HHV-7 was detected in both periodontally diseased and healthy individuals. The prevalence of HHV-6 and -8 was similarly low in both groups. Our data do not support an association of investigated herpesvirus species with destructive periodontal disease. PMID- 12631176 TI - Efficacy of an amine fluoride-triclosan mouthrinse as compared to the individual active ingredients. AB - BACKGROUND: The purpose of the clinical study was to examine the antibacterial and plaque-reducing properties of mouthrinses containing triclosan (TRI), amine fluoride (AmF), and the combination of both (AFT) on 4-day plaque regrowth. A placebo solution (PLA) and a 0.2% chlorhexidine solution (CHX) served as negative and positive controls, respectively. MATERIALS AND METHODS: After a professional tooth cleaning (day 0), 15 volunteers refrained from all mechanical oral hygiene measures for the next 96 h and rinsed instead twice daily for 1 min with 10 ml of one of the five randomly assigned solutions. Plaque index (PlI), which was assessed after 24 and 96 h (PlI1, PlI2), and plaque area of the front teeth (PA), which was planimetrically recorded from disclosed teeth after 96 h, served as clinical parameters. After 24 and 96 h a plaque sample was taken and analyzed microbiologically to evaluate biofilm vitality (VF1, VF2). The subsequent test cycles were conducted after a washout period of 10 days each. RESULTS: No severe adverse events or allergy were seen during the study. CHX influenced all parameters at all time points in comparison to PLA. AFT and AmF showed very similar values (in all parameters), but AmF did not reach the level of significance regarding VF1, nor did AFT with VF2 and PlI1. The TRI solution only reduced PlI2 and PA significantly, but had no influence on biofilm vitality when compared to PLA. CONCLUSION: A synergism between AmF and TRI was not observed. The results suggest that the plaque-reducing and antibacterial effects of the AFT solution are mainly based on the effects of the amine fluoride moiety. PMID- 12631177 TI - Enamel matrix proteins in the treatment of periodontal sites with horizontal type of bone loss. AB - OBJECTIVES: The present study aims to assess the clinical and radiographic outcome of horizontal type of bone loss over a period of 8 months following periodontal surgery with adjunctive use of enamel matrix proteins (EMP). MATERIALS AND METHODS: Twenty patients, who received nonsurgical periodontal therapy and had radiographic horizontal bone loss with an associated probing depth (PD) of > or =4 mm at the maxillary incisor/canine segment, were included. One side of the selected segment divided by the mid-sagittal plane was treated with EMP as part of a crevicular flap. The other side was treated either with a similar intracrevicular (ICI) or a reverse bevel incision (RBI) as part of a conventional flap debridement. Therefore, patients were divided into two groups of 10 patients according to the type of incision performed on the control side. The analysis was based on a classification of two severity groups according to preoperative PD, with the patient's means of measurements for each treatment being the experimental units for the statistical analysis. RESULTS: For pockets of 4-6 mm, EMP treatment was significantly better than the ICI/flap debridement in terms of PD reduction (p<0.001), relative attachment level (RAL) gain (p<0.001) and recession (REC) (p<0.05). Although sites exposed to EMP treatment exhibited significantly greater RAL gain than RBI/flap debridement sites (p<0.01), both treatments resulted in equally effective PD reduction. Less REC occurred with EMP application than with ICI-RBI/flap debridement (p<0.05). Treatment of shallow sites by the conventional flap with both incisions resulted in a tendency for loss of attachment whereas EMP treatment maintained the attachment levels. No significant difference in the degree of probing and radiographic bone levels was found between the treatments. CONCLUSION: EMP treatment showed better clinical improvements as compared to the conventional flap debridement performed with two different incisions. Clinical improvements were more pronounced at periodontal sites with deep, rather than shallow, periodontal pockets. The results of this study provided an important preliminary base for further clinical and histologic studies. PMID- 12631178 TI - Assessment of periodontal conditions and systemic disease in older subjects. AB - BACKGROUND: An increased risk for periodontitis has been associated both with type-1 or insulin dependent diabetes (IDDM) and with type-2 or non-insulin dependent diabetes (NIDDM). AIMS: 1) To describe and analyze periodontal conditions in older low-income ethnic diverse subjects with or without a diagnosis of diabetes. 2) To assess to what extent diabetes mellitus is associated with periodontal status, and 3) how periodontitis ranks as a coexisting disease among other diseases in subjects with diabetes mellitus. MATERIAL AND METHODS: Radiographic signs of alveolar bone loss were studied in 1101 older subjects 60-75 years old (mean age 67.6, SD+/-4.7). The number of periodontal sites and the proportions of teeth with probing depth (PD) > or =5 mm, clinical attachment levels (CAL) > or =4 mm were studied in a subset of 701 of the subjects. RESULTS: IDDM was reported by 2.9% and NIDDM by 9.2% of the subjects. The number of remaining teeth did not differ by diabetic status. The number of sites with PD > or =5 mm and the proportion of PD with > or =5 mm was significantly smaller in the non-diabetic group (chi2=46.8, p<0.01, and chi2=171.1, p<0.001, respectively). Statistical analysis failed to demonstrate group differences for the number and proportions of sites with CAL > or =4 mm and for radiographic findings of alveolar bone loss. Combining all periodontal parameters revealed that the Mantel-Haenszel common odds of having IDDM/NIDDM and periodontitis was 1.8 : 1 (95% CI: 1.1-3.1, p<0.03). The common odds ratio estimate of an association between heart disease and diabetes was 3.6 : 1 (95% CI: 2.1-2.6, p<0.001). CONCLUSIONS: Probing depth differences between IDDM/NIDDM vs. non-diabetic subjects may reflect the presences of pseudo-pockets and not progressive periodontitis in many subjects with diabetes mellitus. Periodontitis is not a predominant coexisting disease in older subjects with diabetes mellitus. PMID- 12631179 TI - Hyperaemic response to cigarette smoking in healthy gingiva. AB - BACKGROUND AND AIM: Cigarette smoking is currently considered as a risk factor for periodontal disease. Controversy exists as to whether the vasoconstrictive property of nicotine is one of the pathogenic mechanisms. To this end we tested the hypothesis that cigarette smoking is causing vasoconstriction in the healthy human gingiva. MATERIALS AND METHODS: Gingival blood flow was continuously measured with laser Doppler flowmetry in healthy (n=13) casual consumers of tobacco. Simultaneously, recordings were made of skin blood flow in the forehead and the thumb as well as heart rate (HR) and blood pressure (BP). In another session infraorbital nerve block anaesthesia (INB) with 1.0 ml of Carbocain without vasoconstrictive additives was used to identify nervously mediated vascular responses to cigarette smoking (n=8). RESULTS: Cigarette smoking induced a modest hyperaemic response in the gingiva that was lower than the relative increases in BP and HR, and the calculated gingival vascular conductance decreased. In the forehead, flow responses were similar to those in the gingiva, while in the thumb a powerful vasoconstriction was observed. During the later part of the 10-min recovery period, BP and HR tended to decrease while blood flow in the gingiva and forehead remained high. INB potentiated the hyperaemic response to cigarette smoking in gingiva. CONCLUSIONS: The present results help to shed some light on the understanding of the vasoactive mechanisms induced by cigarette smoking, and to support the hypothesis that cigarette smoking causes nervously mediated vasoconstriction in the healthy human gingiva. However, the degree of vasoconstriction was far less than in the thumb skin, and in our subjects was overcome by the evoked rise in arterial perfusion pressure. As a consequence, gingival blood flow increased during smoking. It is speculated that small repeated vasoconstrictive attacks due to cigarette smoking may in the long run contribute to gingival vascular dysfunction and periodontal disease. PMID- 12631180 TI - Change of antibiotic susceptibility following periodontal therapy. A pilot study in aggressive periodontal disease. AB - BACKGROUND: The hypothesis was tested that bacterial susceptibilities in aggressive periodontitis change upon administration of systemic antibiotics as adjuncts to periodontal therapy. METHODS: In 23 subjects (average age 38.9+/-6.7 years) with aggressive periodontitis, microbial parameters were assessed prior to and 1 year after completion of comprehensive mechanical/surgical and systemic antimicrobial therapy. Following identification of five selected pathogens with the Rapid ID 32 A system, their susceptibilities towards amoxicillin/clavulanate potassium, metronidazole, and tetracycline were examined with the E-test. Antibiotics were administered according to the test results, and the minimal inhibitory concentrations (MIC90) were reevaluated after 1 year. Statistical analysis was performed on a patient basis, with the site data used for evaluation of the MIC levels. RESULTS: Bacterial MIC levels remained constant among the three antibiotic treatment groups compared with baseline. Mean MIC90 values ranged from <0.02 to 0.11 microg/ml (amoxicillin/clavulanate potassium), <0.02 to 0.27 microg/ml (metronidazole), and <0.02 to 0.11 microg/ml (tetracycline). Observed changes in susceptibility were attributed to the elimination of single bacterial taxa in the subgingival environment after antibiotic therapy. There were no statistically significant differences in clinical parameters among the treatment groups. Single tetracycline MICs were 1.5- to 6-fold enhanced compared to amoxicillin/clavulanate potassium and metronidazole. CONCLUSION: The periodontal pathogens investigated prior to and 1 year after periodontal therapy are tested sensitive to the antimicrobial agents. In aggressive periodontitis, changes in bacterial susceptibility upon the administration of systemic antibiotics are associated with the limited number of isolates tested following therapy. PMID- 12631181 TI - Epidemiologic risk factors for periodontal attachment loss among adults in the United States. AB - OBJECTIVES: The objective of this study was to identify variables related to periodontal loss of attachment (LOA). MATERIALS AND METHODS: The study population consisted of a nationally representative sample of 12,325 US adults who participated in the National Health and Nutrition Examination Survey III. The analyses used survey multinomial logistic regression to account for the complex survey design. RESULTS: The adjusted odds ratio for a mean LOA of 3 mm or more associated with current smoking was 18.55 (95% CI 9.44-36.45) among 20-49-year olds. Among those aged 50 years or more, the odds ratio for a mean LOA of 4 mm or more was 25.64 (13.04-50.40). Prior smoking, untreated decayed surfaces (on both person and site levels), and male gender were also associated with LOA. There was no excess risk observed among non-Hispanic blacks or Mexican-Americans. The adjusted population attributable fractions due to current smoking were 60% for persons aged 20-49 with the worst 10% of LOA (1.58 mm or more), and 47% for those aged 50 plus (LOA of 3.39 mm or more). CONCLUSION: These results support earlier findings regarding the central role of cigarette smoking in the etiology of periodontal loss of attachment, a role due in large part to the substantial relationship between smoking and severe periodontal disease. PMID- 12631182 TI - Effects of plaque, psychological stress and gender on crevicular Il-1beta and Il 1ra secretion. AB - OBJECTIVES: The study aimed to analyse (a) whether the effects of psychological stress and of experimental gingivitis on interleukin-1beta (Il-1beta) described before are compensated by concomitant increases in Il-1 receptor antagonist (Il 1ra), and (b) whether there do exist any gender differences in the Il-1 responses to experimental gingivitis and to psychological stress. MATERIALS AND METHODS: Thirteen medical students participating in a major academic exam (seven males, six females) and 14 medical students without academic stress (eight males, six females) refrained from oral hygiene in two antagonistic quadrants for 28 days (plaque) while they maintained oral hygiene in the remaining quadrants (hygiene). Weekly crevicular fluid samples of plaque and hygiene sites were assayed for Il 1beta and Il-1ra. RESULTS: Neither stress nor experimental gingivitis exerted significant effects on Il-1ra. In controls, we observed significant gender and gender x time effects on Il-1beta; comparing stress groups, gender x time and stress x gender x time interactions became significant. Women show a reduced Il 1beta response to plaque at rest and an increased response under stress. Similar results were found with respect to bleeding on probing. CONCLUSIONS: Gender must be controlled in studies on periodontal responses to pathogens. Stress plays a role in these responses. PMID- 12631183 TI - Association of the -1087 IL 10 gene polymorphism with severe chronic periodontitis in Swedish Caucasians. AB - BACKGROUND: Severe forms of periodontitis are suggested to have a genetic basis. OBJECTIVE: The aim of the present investigation was to study association of an IL10 gene polymorphism (G to A transition at the -1087 position) with severe chronic periodontitis. MATERIALS AND METHODS: Two groups of Swedish Caucasian subjects were included. One group consisted of 60 patients (aged 36-74 years; mean 54.5+/-8.5) with severe and generalized chronic periodontitis. The patients exhibited bone loss >50% at all teeth. Thirty-nine periodontally healthy subjects between 35-78 years of age (mean 51.0+/-10.9) were also recruited. DNA was isolated from peripheral blood cells and genotyping was performed by combination of PCR with restriction endonuclease mapping. RESULTS: The proportion of subjects that exhibited the GG genotype was significantly larger in the group with severe periodontitis than in the periodontally healthy group. The difference regarding the occurrence of the GG genotype between the two groups was more conspicuous in non-smokers and yielded an odds ratio of 6.1. The G allele carriage in non smokers was >90 % in the periodontitis group and was significantly higher than in the healthy controls. CONCLUSION: It is suggested that the -1087 IL10 polymorphism in Caucasian subjects of a north European origin is associated with severe chronic periodontitis. PMID- 12631184 TI - Effect of toothpaste on the plaque inhibitory properties of a cetylpyridinium chloride mouth rinse. AB - BACKGROUND AND AIMS: Cetylpyridinium chloride (CPC) mouth rinses have moderate plaque inhibitory activity when used alone but rarely have shown adjunctive benefits to tooth brushing with toothpaste. Several explanations for this apparent anomaly can be proferred, including loss of antiseptic activity due to interactions with toothpaste ingredients. The aim of this study was to measure the effect of toothpaste on the plaque inhibitory properties of a CPC mouth rinse using paired rinses of CPC, toothpaste slurry (TP) and water (W). METHODS: The study was a single blind, randomised, seven-treatment, cross over design balanced for residual effects, involving 21 healthy, dentate subjects. The paired rinses were: W-CPC, CPC-W, TP-CPC, CPC-TP, W-TP, TP-W and W-W. Rinsing with solutions or slurries was done for 60 s twice per day. On day 1, subjects were rendered plaque free, suspended tooth cleaning and commenced the allocated rinse regimen. On day 5, plaque was scored by index. A 2(1/2) day wash out of normal oral hygiene was allowed between each regimen. RESULTS: The order from lowest to highest plaque scores was as follows: W-CPC = CPC-W < CPC-TP < TP-CPC < or = W-TP < TP-W < W-W. Several differences in pairs of treatments were statistically significant, the most relevant of which were significantly less plaque with W-CPC compared to TP CPC, TP-W and W-TP, and significantly more plaque with W-W compared to all other regimens except TP-W. CONCLUSIONS: Toothpaste, whilst possessing some plaque inhibitory activity, when used immediately before a CPC mouth rinse adversely affected the plaque inhibitory action of this antiseptic. This in part may explain the reported lack of adjunctive benefits of CPC rinses to normal oral hygiene practices and supports the suggestion, made for chlorhexidine rinses, that their use should follow toothpaste by at least 60 min. PMID- 12631185 TI - Deproteinized cancellous bovine bone (Bio-Oss) as bone substitute for sinus floor elevation. A retrospective, histomorphometrical study of five cases. AB - OBJECTIVES: To study in detail the performance of deproteinized cancellous bovine bone (DPBB, Bio-Osso) granules as a bone substitute, a histomorphometric was performed on five patients treated with DPBB for reconstruction of the severely atrophic maxilla. MATERIAL AND METHODS: DPBB was used as mixture with autogenous bone particles, in concentrations that increased from 20% to 100% DPBB, with the time of healing increasing accordingly from 5 to 8 months. A total of 20 vertical biopsies was taken at the time of fixture installation and used for histomorphometry as undecalcified Goldner stained sections. RESULTS: The results show that in all cases, the DPBB granules had been interconnected by bridges of vital newly formed bone. The volume of bone in the grafted area correlated inversely with the concentration of DPBB grafted, and varied between 37% and 23%. However, the total volume of mineralized material (bone plus DPI3B granules) remained within the same range in all five patients (between 53% and 59%). The high values for osteoid and resorption surface, and the presence of tartrate resistant acid phosphatase-positive multinucleated osteoclasts in resorption lacunae, indicated that bone remodeling was very active in all grafts. Osteoclasts were also observed in shallow resorption pits on DPBB surfaces. The percentage DPBB surface in contact with bone remained stable at about 35% and could not be related to the proportion of DPBB grafted. CONCLUSION: Although the number of patients examined was limited, the data suggest that deproteinized cancellous bovine bone, preferably combined with autogenous bone particles, is a suitable material for sinus floor elevation in the severely atrophic human maxilla. PMID- 12631186 TI - C-1 esterase inhibitor dysfunction localised to the periodontal tissues: clues to the role of stress in the pathogenesis of chronic periodontitis? AB - BACKGROUND: C1-esterase inhibitor (C1eIn) is an important modulator of complement activation via the classical pathway. Deficiencies or dysfunction involving this inhibitor underlie the condition of angioneurotic oedema. AIM: The purpose of this report is to describe a female patient who presented at the age of 24 years with an apparently aggressive form of periodontitis and severe oedema, localised to the free gingival tissues. After 21 years of repeated surgical reduction of the gingiva, a diagnosis of C1eIn dysfunction was made. METHODS: Exhaustive serological investigations were performed along with histopathology. RESULTS: All investigations were unremarkable, until the function of the C1eIn molecule was investigated. These demonstrated a functional activity of only 29% and a raised C1q at 157 mg/l. Subsequent repeated investigation with careful specimen handling demonstrated undetectable levels of C1eIn and normal C1q. A diagnosis of C1eIn dysfunction was made, although at present it is unclear whether this represents an unusual variant of hereditary dysfunctional C1eIn deficiency. The patient was managed by various means, including steriodal and non-steroidal drugs, the latter forming part of her maintenance regime. CONCLUSIONS: To our knowledge, this is the first case of angio-oedema localised to the free gingiva. The role of stress in the acute exacerbations of oedema and bone loss is discussed along with the diagnostic pitfalls associated with this case. PMID- 12631188 TI - New frontiers in biological halogenation. PMID- 12631189 TI - Effect of water activity and temperature on germination and growth of Penicillium digitatum, P. italicum and Geotrichum candidum. AB - AIMS: This study compares the effect of temperature (4-37 degrees C) and water activity (aw: 0.99-0.87) and their interactions on the germination rates, lag times prior to germination and mycelial growth 'in vitro' of Penicillium digitatum, P. italicum and Geotrichum candidum, the main postharvest pathogens affecting citrus fruits. METHODS AND RESULTS: Germination and growth were markedly influenced by temperature and aw. Generally, lag times were longer and germination and growth rates were slower when conditions of temperature and aw were far from optimum. All the studied species were able to germinate over a range of 4-30 degrees C at 0.995 aw, although in non-optimal conditions P. digitatum only reached 40-60% of germinated conidia. At low temperatures, P. italicum germinated and grew faster than P. digitatum and G. candidum, particularly at 0.95 aw. Penicillium italicum was also able to germinate and grow in the driest studied conditions (0.87 aw), while G. candidum did not germinate under 0.95 aw. CONCLUSIONS: Knowledge of the ecological requirements of these fungi is important in order to understand their behaviour in natural situations and to predict fungal spoilage on citrus fruits. PMID- 12631190 TI - The adaptive response of Escherichia coli O157 in an environment with changing pH. AB - AIMS: To predict and validate survival of non-acid adapted Escherichia coli O157 in an environment mimicking the human stomach. METHODS AND RESULTS: Survival was predicted mathematically from inactivation rates at various, but constant pH values. Predictions were subsequently validated experimentally in a pH-controlled fermentor. Contrary to prediction, acid-sensitive cultures of E. coli O157 survived for a long period of time and died as rapidly as acid-resistant cultures. Experimental results showed that in an environment with changing pH, acid-sensitive cultures became acid-resistant within 17 min. Cyclo fatty acids was reported to be a factor in acid resistance. As synthesis of cyclo fatty acids does not require de novo enzyme synthesis and thus requires little time to develop, we analysed the membrane fatty acid composition of E. coli O157 during adaptation. No changes in membrane fatty acid composition were observed. CONCLUSIONS: Acid adaptation of E. coli O157 can occur during passage of the human gastric acid barrier, which can take up to 4 h. SIGNIFICANCE AND IMPACT OF THE STUDY: The ability of acid-adapted bacteria to survive the human stomach is an important virulence factor. The ability of non-acid adapted E. coli O157 to adapt within a very short period of time under extreme conditions further contributes to the virulence of E. coli O157. PMID- 12631191 TI - Autolytic activity and pediocin-induced lysis in Pediococcus acidilactici and Pediococcus pentosaceus strains. AB - AIMS: To evaluate the autolytic phenotype of Pediococcus acidilactici and P. pentosaceus, the peptidoglycan hydrolases content and the effect of pediocin AcH/PA-1 and autolysins on cell lysis. METHODS AND RESULTS: The autolytic phenotype of Pediococcus strains was evaluated under starvation conditions in potassium phosphate buffer. The strains tested showed an extent of autolysis ranging between 40 and 90% after 48 h of starvation at 37 degrees C. Peptidoglycan hydrolase content was evaluated by renaturing sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) using cells of Micrococcus lysodeikticus as a target for the enzymatic activity and a major activity band migrating at about 116 kDa was detected. Additional secondary lytic bands migrating in a range of molecular weight between 45 and 110 kDa were also detected. The lytic activity, evaluated in the presence of different chemicals, was retained in 15 mM CaCl2 and in a range of pH between 5 and 9.5 but was strongly reduced in presence of 8% NaCl and in the presence of protease inhibitors. The substrate specificity of peptidoglycan hydrolases of Pediococcus strains was evaluated in renaturing SDS-PAGE incorporating cells of different bacterial species. Lytic activity was detected against cells of Lactococcus lactis subsp. lactis, L. delbrueckii subsp. bulgaricus, Lactobacillus helveticus and Listeria monocytogenes. The interaction between pediocin AcH/PA-1 and autolysis was evaluated and a relevant effect of bacteriocin in cell-induced lysis was observed. CONCLUSIONS: The autolytic phenotype is widely distributed among P. acidilactici and P. pentosaceus and the rate of autolysis is high in the majority of the analysed strains. Several autolytic bands, detected by renaturing SDS-PAGE, retained their activity against several lactic acid bacteria and L. monocytogenes. SIGNIFICANCE AND IMPACT OF THE STUDY: The characterization of the autolytic phenotype of Pediococcus acidilactici and P. pentosaceus strains should expand the knowledge of their role in fermentation processes where these species occur as primary or secondary bacterial population. PMID- 12631192 TI - Modelling and optimization of inactivation of Lactobacillus plantarum by pulsed electric field treatment. AB - AIMS: The effect of critical pulsed electric field (PEF) process parameters, such as electric field strength, pulse length and number of pulses, on inactivation of Lactobacillus plantarum was investigated. METHODS AND RESULTS: Experiments were performed in a pH 4.5 sodium phosphate buffer having a conductivity of 0.1 S m-1, using a laboratory-scale continuous PEF apparatus with a co-linear treatment chamber. An inactivation model was developed as a function of field strength, pulse length and number of pulses. Based on this inactivation model, the conditions for a PEF treatment were optimized with respect to the minimum energy required to obtain a certain level of inactivation. It was shown that the least efficient process parameter in the range investigated was the number of pulses. The most efficient way to optimize inactivation of Lact. plantarum was to increase the field strength up to 25.7 kV cm-1, at the shortest pulse length investigated, 0.85 micros, and using a minimum number of pulses. The highest inactivation of Lact. plantarum at the lowest energy costs is obtained by using the equation: E=26.7tau0.23, in which E is the field strength and tau the pulse length. An optimum is reached by substituting tau with 5.1. CONCLUSIONS: This study demonstrates that the correct choice of parameters, as predicted by the model described here, can considerably improve the PEF process. SIGNIFICANCE AND IMPACT OF THE STUDY: The knowledge gained in this study improves the understanding of the limitations and opportunities of the PEF process. Consequently, the advantage of the PEF process as a new option for non-thermal decontamination can be better utilized. PMID- 12631193 TI - Improved methods of cultivation and production of deuteriated proteins from E. coli strains grown on fully deuteriated minimal medium. AB - AIMS: The aim was to develop reliable and economical protocols for the production of fully deuteriated biomolecules by bacteria. This required the preparation of deuterium-tolerant bacterial strains and an understanding of the physiological mechanisms of acquisition of deuterium tolerance. METHODS AND RESULTS: We report here improved methods for the cultivation of Escherichia coli on fully deuteriated minimal medium. A multi-stage adaptation protocol was developed; this included repeated plating and selection of colonies and resulted in highly deuterium-tolerant cell cultures. Three E. coli strains, JM109, MRE600 and MRE600Rif, were adapted to growth on deuteriated succinate medium. This is the first report of JM109 being adapted to deuteriated minimal media. The adapted strains showed good, consistent growth rates and were capable of being transformed with plasmids. Expression of heterologous proteins in these strains was reliable and yields were consistently high (100-200 mg l-1). We also show that all E. coli cells are inherently capable of growth on deuteriated media. CONCLUSIONS: We have developed a new adaptation protocol that resulted in three highly deuterium-tolerant E. coli strains. Deuterium-adapted cultures produced good yields of a deuteriated recombinant protein. We suggest that E. coli cells are inherently capable of growth on deuteriated media, but that non-specific mutations enhance deuterium tolerance. Thus plating and selection of colonies leads to highly deuterium-tolerant strains. SIGNIFICANCE AND IMPACT OF STUDY: An understanding of the mechanism of adaptation of E. coli to growth on deuteriated media allows strategies for the development of disabled deuterium-tolerant strains suitable for high-level production of deuteriated recombinant proteins and other biomolecules. This is of particular importance for nuclear magnetic resonance and neutron scattering studies of biomolecules. PMID- 12631194 TI - Development of a 5'-nuclease PCR assay for detecting Shiga toxin-producing Escherichia coli O145 based on the identification of an 'O-island 29' homologue. AB - AIMS: A DNA sequence, from Escherichia coli STEC O145, homologous to O-island 29 from STEC O157 is described, together with a real-time PCR assay for detecting it. METHODS AND RESULTS: PCR and sequencing were used to identify the 'O-island 29' homologous DNA sequence from STEC O145 (strain VTH34). The sequence divergence between the STEC O145 and O157 'O-island 29' allowed a STEC O145 5' nuclease PCR assay to be developed. CONCLUSIONS: The characterization of a novel locus in STEC O145 has allowed a specific O145 serogroup 5'-nuclease PCR assay to be designed. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings increase the number of serogroup PCR assays available as alternatives to classical O serotyping of E. coli. PMID- 12631195 TI - Genotypic and phenotypic characterization of the dynamics of the lactic acid bacterial population of adjunct-containing Cheddar cheese manufactured from raw and microfiltered pasteurised milk. AB - AIMS: This study investigates the dynamics of the microflora, particularly the lactobacilli, in Cheddar cheese manufactured from raw and microfiltered milk containing different adjunct cultures. METHODS AND RESULTS: Sixteen cheeses - raw milk, adjunct and control cheeses - were manufactured in four trials. Lactobacilli were identified by PCR methods in one trial, and by phenotypic typing for all trials. Numbers of lactobacilli were significantly different at day 1 and 3 months in the control and adjunct-containing cheeses. In the raw milk cheeses, Lactobacillus paracasei was detected throughout ripening, Lact. curvatus at the end, and Lact. plantarum at day 1 only. Lactobacillus strain diversity decreased from raw, control to adjunct cheeses. Enteroccoci and coliform numbers further differentiated raw cheeses from the others. Lactococcal starter numbers also differed in the three cheese types and differences were observed within adjunct cheeses. Although adjunct lactobacilli dominated in the cheese to which they were added, strains with similar phenotypic profiles were also detected on occasions in some of the control cheeses. CONCLUSIONS: The addition of adjunct lactobacilli modified the growth kinetics of both adventitious lactobacilli and starter lactococci during ripening. Appropriate strain tracking is necessary to monitor changes in the population profiles of control and experimental cheeses in trials utilizing adjunct cultures. SIGNIFICANCE AND IMPACT OF THE STUDY: Investigations of the role of adjunct strain(s) in cheeses may be complicated by the interactions between the adjunct and the other cheese strains, and effective strain monitoring by genotypic or phenotypic methods is essential if valid comparisons are to be made. PMID- 12631196 TI - Effects of carbon substrate enrichment and DOC concentration on biodegradation of PAHs in soil. AB - AIMS: Two common reasons to explain slow environmental biodegradation of polycyclic aromatic hydrocarbons (PAHs), namely lack of appropriate carbon sources for microbial growth and limited bioavailability of PAHs, were tested in a laboratory bioassay using a creosote-contaminated soil. METHODS AND RESULTS: The soil, containing a total of 8 mg g-1 of 16 PAHs, was sieved and incubated in bottles for 45 days. The first explanation was tested by enrichment with the analogue anthracene and the non-analogue myristic acid, and both failed to stimulate degradation of all PAHs except anthracene. The second explanation was tested by addition of different concentrations of dissolved organic carbon (DOC), with effects depending on the DOC concentration and the molecular size of the PAH. The degradation was enhanced from 10 to 35% for 12 PAHs when the soil was saturated. The degraded amounts of individual PAHs were proportional to their concentration in the soil. CONCLUSIONS: The slow in situ degradation of PAHs was enhanced by more than three times by adding water as a solvent. Addition of DOC facilitated the degradation of four- to six-ring PAHs. SIGNIFICANCE AND IMPACT OF STUDY: Bioremediation of PAH-contaminated sites may be facilitated by creating water-saturated conditions but retarded by addition of other carbon substrates, such as analogue compounds. PMID- 12631197 TI - Overproduction of laccase by a monokaryotic strain of Pycnoporus cinnabarinus using ethanol as inducer. AB - AIMS: Laccase production by the monokaryotic strain Pycnoporus cinnabarinus ss3 was studied using ethanol as inducer in the culture medium. METHODS AND RESULTS: The effect of ethanol was tested at 10, 20, 30, 35 and 45 g l-1 and compared with that of ferulic acid, known until now as the most efficient inducer for laccase expression by P. cinnabarinus ss3. In the presence of 35 g l-1 ethanol, laccase activity (266 600 U l-1) and productivity (19 000 U l-1 day-1) were nine and fivefold higher compared with ferulic acid-induced cultures, and 155- and 65-fold higher compared with non-induced cultures, respectively. In vivo, ethanol added to the culture medium of P. cinnabarinus ss3 favoured a continuous and high expression of laccase gene. Under these conditions, P. cinnabarinus ss3 produced preferentially the isoenzyme LAC I. Ethanol added in vitro to the purified P. cinnabarinus ss3 laccase typically inhibited the enzymatic activity. CONCLUSIONS: In spite of an initial inhibitory effect on mycelial growth, ethanol was shown to be a very strong inducer for laccase expression by P. cinnabarinus ss3 allowing an average yield of 1-1.5 g l-1 laccase. SIGNIFICANCE AND IMPACT OF THE STUDY: This study identified P. cinnabarinus ss3 as an outstanding producer of laccase in the presence of ethanol as inducer. Ethanol is an inexpensive agricultural by product and the process is simple to scale-up for industrial production. PMID- 12631198 TI - Adaptive responses of Salmonella enterica serovar Typhimurium DT104 and other S. Typhimurium strains and Escherichia coli O157 to low pH environments. AB - AIMS: Cattle are a known main reservoir for acid-resistant Escherichia coli O157 and Salmonella enterica serovar Typhimurium DT104. We studied the response of S. Typhimurium DT104 to extreme low pH environments and compared their response to that of acid-resistant E. coli O157 and other S. Typhimurium phage types. METHODS AND RESULTS: Bacteria were grown in nutrient-rich medium and subsequently acid challenged at pH 2.5. We found that stationary phase cultures of various S. Typhimurium strains were able to survive a challenge for 2 h at pH 2.5. As in E. coli, the ability of S. Typhimurium to survive at pH 2.5 was shown to be dependent on the presence of amino acids, specifically arginine. The amount of proton pumping H+/ATPase, both in E. coli O157 and S. Typhimurium strains, was lower when grown at pH values <6 than after growth at pH 7.5. Cyclo fatty acid content of membranes of bacteria grown at pH values <6 was higher than that of membranes of bacteria grown at pH 7.5. CONCLUSIONS: Various S. Typhimurium strains, both DT104 and non-DT104, are able to survive for a prolonged period of time at pH 2.5. Their response to such low pH environment is seemingly similar to that of E. coli O157. SIGNIFICANCE AND IMPACT OF THE STUDY: Food-borne pathogens like S. Typhimurium DT104 and E. coli O157 form a serious threat to public health since such strains are able to survive under extreme low pH conditions as present in the human stomach. The emergence these acid-resistant strains suggests the presence of a selection barrier. The intestinal tract of ruminants fed a carbohydrate-rich diet might be such a barrier. PMID- 12631200 TI - Characterization of sourdough lactic acid bacteria based on genotypic and cell wall protein analyses. AB - AIMS: To evaluate the effectiveness of two independent methods in differentiating a large population of lactic acid bacteria (LAB) isolated from wheat flours and sourdoughs and to correlate eventual differences/similarities among strains with their geographical origin and/or process parameters. METHODS AND RESULTS: One hundred fifty strains belonging to Lactobacillus spp. and Weissella spp., plus eight type strains, one for each species, and two unidentified isolates, were characterized by randomly amplified polymorphic DNA (RAPD) and SDS-PAGE of cell wall proteins. The RAPD analysis separated the eight type strains but did not always assign all the strains of a species to the same group, while SDS-PAGE cell wall protein profiles were species-specific. Frequently, strains isolated from sourdoughs of the same geographical origin or produced by similar raw material/process parameters showed similar RAPD and/or cell-wall profiles. CONCLUSIONS: The combined use of the RAPD and cell-wall protein analysis represents a useful tool to classify large adventitious microbial populations and to discriminate the diversity of the strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This study represents a typing of a large collection of flour/sourdough LAB and provides evidence of the advantage of using two independent methods in the classification and traceability of microorganisms. PMID- 12631199 TI - Molecular epidemiological survey of Listeria monocytogenes in broilers and poultry products. AB - AIMS: To investigate the prevalence of Listeria monocytogenes in poultry products, and to elucidate whether poultry products may be linked to listeriosis cases. A further goal was to identify contamination routes for L. monocytogenes to broiler carcasses. METHODS AND RESULTS: Poultry products (385 samples) were screened for L. monocytogenes. The recovered isolates and 19 patient isolates were characterized by multilocus enzyme electrophoresis and restriction enzyme analysis. The poultry isolates showed great genetic diversity, but no identical subclones were identified from poultry sources and patients. One slaughterhouse was examined in detail during a 16-month period. The contamination rates increased along the processing line, and one subclone was found during the whole period. Only low prevalence of the bacteria was revealed from broiler faeces. CONCLUSIONS: The prevalence of L. monocytogenes in poultry products was high, but no listeriosis cases was linked to poultry products. Broilers seem to be contaminated during the slaughter process, and specific strains may persist in the processing environment. Broiler faeces does not seem to be an important source of L. monocytogenes in poultry products. SIGNIFICANCE AND IMPACT OF THE STUDY: Preventive measures to avoid contamination of poultry products by L. monocytogenes must be taken in the processing plants. PMID- 12631201 TI - Growth and molecular characterization of dental plaque microcosms. AB - AIMS: (i) To compare the effects of feeding protocols upon the composition and stability of dental plaque microcosms formed in constant-depth film fermenters (CDFF). (ii) To evaluate the utility of denaturing gradient gel electrophoresis (DGGE) and culture methodologies for the investigation of such models. METHODS AND RESULTS: Microcosms were established anaerobically in the CDFFs from freshly collected saliva. These were fed either with artificial saliva alone (famine) or combined with discontinuous feeding (feast-famine). Culture and 16s rDNA sequencing indicated that supplemental feeding gave ca. 2 log increases in Lactobacillus rhamnosus and Prevotella buccae. Feast-famine microcosms were then further characterized by DGGE using primers specific for the V2-V3 region of eubacterial rDNA. These gave single major bands with pure cultures (eight species) and resolved all strains apart from Lact. rhamnosus and Actinomyces naeslundii. Whilst culture with selective media indicated a degree of stability and reproducibility between replicate microcosms, DGGE showed a considerable degree of variability that related to several putatively uncultured bacteria. CONCLUSIONS: Feast-famine regimes altered community composition. DGGE analyses identified putatively unculturable species and demonstrated variability between replicate fermenters. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the utility of DGGE for the analysis of dental plaque, especially with respect to unculturable bacteria. Results question the assumptions of reproducibility of plaque microcosms established in non-replicated CDFFs made on the basis of selective media. Feeding regimes, particularly those involving complex nutrients, will dramatically affect population dynamics. PMID- 12631202 TI - Antifungal activity of essential oils evaluated by two different application techniques against rye bread spoilage fungi. AB - AIMS: To study how antifungal activity of natural essential oils depends on the assay method used. METHODS AND RESULTS: Oils of bay, cinnamon leaf, clove, lemongrass, mustard, orange, sage, thyme and two rosemary oils were tested by two methods: (1) a rye bread-based agar medium was supplemented with 100 and 250 microl l-1 essential oil and (2) real rye bread was exposed to 136 and 272 microl l-1 volatile oil in air. Rye bread spoilage fungi were used for testing. Method 1 proved thyme oil to be the overall best growth inhibitor, followed by clove and cinnamon. On the contrary, orange, sage and rosemary oils had very limited effects. Mustard and lemongrass were the most effective oils by the volatile method, and orange, sage and one rosemary showed some effects. Oil compositions were analysed by gas chromatography-mass spectrography. CONCLUSIONS: Antifungal effects of the essential oils depended on the application method. Larger phenolic compounds such as thymol and eugenol (thyme, cinnamon and clove) had best effect applied directly to medium, whereas smaller compounds such as allyl isothiocyanate and citral (mustard and lemongrass) were most efficient when added as volatiles. SIGNIFICANCE AND IMPACT OF THE STUDY: This study proves that the method used for screening essential oils as potential antimicrobials should correspond with the application sought. PMID- 12631203 TI - Utilization of carbonate and ammonia-based treatments to eliminate Escherichia coli O157:H7 and Salmonella Typhimurium DT104 from cattle manure. AB - AIMS: The objective of this study was to investigate alkaline treatments of cattle manure to kill coliforms, Escherichia coli O157:H7 and Salmonella Typhimurium DT104 based on their inhibition by carbonate ion and ammonia. METHODS AND RESULTS: Pure cultures of S. Typhimurium DT104 and E. coli O157:H7 strains were treated with sodium carbonate and ammonia to determine threshold inhibitory concentrations. Fresh cattle manure samples were inoculated with the same strains and their survival was determined after addition of sodium hydroxide, ammonium sulphate, sodium carbonate and/or urea. Control of CO and NH3 concentrations in manure by pH adjustment to 9.5 with sodium hydroxide to more than 5 and 30 mmol l 1, respectively, killed more than 106 cells g-1 in 7 days. Addition of sodium carbonate enhanced the killing effect of NaOH by increasing the CO and NH3 concentrations. Addition of 100 mmol l-1 urea, produced high levels of CO and NH3 and decreased all bacterial counts by at least 106 cells g-1 after 7 days. CONCLUSIONS: Reduction of food-borne pathogens in manure can be achieved by a combination of high concentrations of CO and NH3 which are pH-dependent parameters. SIGNIFICANCE AND IMPACT OF STUDY: Addition of urea could provide a simple manure treatment by combining both antimicrobial factors. PMID- 12631204 TI - Mutations in the genes for synthesis of the outer core region of the lipopolysaccharide of Yersinia enterocolitica O:3. AB - AIMS: The aim of this study was to construct non-polar frame-shift mutations in some of the individual genes responsible for the biosynthesis of the branching outer core (OC) hexasaccharide of the lipopolysaccharide (LPS) in Yersinia enterocolitica O:3 (YeO:3). METHODS AND RESULTS: Chromosomal segments of YeO:3 containing wbcN, wbcO and wbcQ genes were cloned into a suicide vector. A frame shift mutation was introduced into each gene by modifying a unique restriction enzyme recognition site. Each recombinant plasmid with a modified OC gene was mobilized into YeO:3 to allow for allelic exchange between the modified gene and the wild type chromosomal gene. The exchange was confirmed by demonstrating the absence of the particular restriction site in the chromosome of each mutant strain. Analysis of LPS by gel electrophoresis showed that the LPS of the mutants was lacking the OC. Therefore, the constructed wbcN, wbcO and wbcQ strains are true mutants with frame-shifts in the corresponding genes. CONCLUSIONS: The products of the wbcN, wbcO and wbcQ genes are putative glycosyltransferases and, based on the present analysis, essential for the biosynthesis of the OC hexasaccharide. The absence of OC in the LPS of these mutants further supports the hypothesis that the OC hexasaccharide is a single O-antigen O-unit that is not polymerized in YeO:3. SIGNIFICANCE AND IMPACT OF THE STUDY: These mutants provide information on the unique nature of the synthesis of OC of YeO:3 LPS. They are valuable for future biochemical studies to establish the roles of the products of individual OC genes. PMID- 12631205 TI - The prevalence of Salmonella spp. in bovine faecal, rumen and carcass samples at a commercial abattoir. AB - AIMS: To determine the prevalence, serotype and antibiotic resistance profile of Salmonella isolates in cattle and on carcasses at a commercial Irish abattoir. METHODS AND RESULTS: Faecal, rumen and carcass samples were collected from a beef abattoir over a 12-month period and examined for the presence of Salmonella spp. Isolates were serotyped, phage typed (when serotype was found to be S. Typhimurium) and tested for susceptibility to a panel of antibiotics. Salmonella was isolated from 2% of faecal, 2% of rumen and 7.6% of carcass samples. Salmonella was most frequently isolated from samples taken during the period August to October. S. Dublin was isolated from 72% of positive samples. S. Agona and S. Typhimurium definitive type (DT)104 were each isolated from 14% of positive samples. All S. Typhimurium DT104 isolates were resistant to ampicillin, chloramphenicol, streptomycin, sulphafurazole and tetracycline (ACSSuT). On occasion, from a single animal, the same serotype was isolated from more than one sample (i.e. faeces and rumen; faeces and carcass; rumen and carcass; faeces, rumen and carcass). CONCLUSIONS: Salmonella is present in cattle at slaughter and on beef carcasses at an Irish abattoir, with a higher frequency of occurrence during the period August to October. Most isolates from the study are not commonly associated with human clinical infection, with the exception of S. Typhimurium DT104 (R-type ACSSuT). SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides epidemiological data that is necessary for the understanding of beef as a source of human Salmonella infection. PMID- 12631206 TI - Aerotolerance of human clinical isolates of Prevotella spp. AB - AIMS: To determine the influence of oxygen exposure and growth stages on oxygen tolerance for clinical and reference specimens of the genus Prevotella. METHODS AND RESULTS: Tolerance to oxygen exposure was evaluated along growth stages for a total of four Prevotella isolates constituted by two strains tested soon after isolation from periodontally compromised patients and two reference strains kept frozen (-86 degrees C) in our laboratory collection. Additionally, the recently recovered isolates were also assayed after 4 months exposed to oxygen during laboratory handling. On solid medium, recently recovered Prevotella specimens proved to be less tolerant to oxygen exposure soon after isolation than when exposed for 4 months to oxygen during laboratory handling. Oxygen resistance of reference strains maintained in the laboratory collection showed to be the highest when compared with the clinical isolates both before and after oxygen exposure. When oxygen tolerance was tested during growth, bacterial cells from the exponential phase were more tolerant. Differences were observed in protein patterns between clinical isolates soon after recovery and after laboratory handling as determined by SDS-PAGE of crude cell-free extracts. However, SOD activities were similar in all bacterial samples tested. CONCLUSIONS: The ability to adaptively change oxygen tolerance was observed for pathogenic specimens of Prevotella. This property may be considered a virulence factor. SIGNIFICANCE AND IMPACT OF THE STUDY: Adaptative aerotolerance of Prevotella as a factor for virulence and survival. PMID- 12631208 TI - Subtyping Listeria monocytogenes through the combined analyses of genotype and expression of the hlyA virulence determinant. AB - AIMS: A major challenge for Listeria monocytogenes diagnostics is that this bacterium is ubiquitous in the environment, and that only a small fraction of the lineages are potential human pathogens. The aim of this work was to obtain a better subtyping of L. monocytogenes through utilization of combined analyses of genotype and the expression of the virulence determinant hlyA. METHODS AND RESULTS: We investigated the effect of growth temperature and medium on the hlyA expression. The gene expression levels were determined by real-time quantitative reverse transcription PCR. The expression pattern of hlyA was highly diverse among the different strains tested. The expression ranged from repression to a 1000-fold induction for growth at 42 degrees C, as compared with 0 degrees C. The expression patterns were compared with the corresponding genotypes. There were surprisingly low correlations between the expression patterns and the genotype clusterings. This is exemplified for the virulent type strain NTNC 7973 and non virulent type strain DSMZ 20600. These strains are genetically nearly identical, while the hlyA gene expression patterns are very different. CONCLUSIONS: The hlyA gene expression was highly diverse even within genetically clustered subgroups of L. monocytogenes. Consequently, the gene expression patterns can be used to further differentiate the strains within these genetic subgroups. SIGNIFICANCE AND IMPACT OF THE STUDY: A major limitation in the control of L. monocytogenes is that the current tools for subtyping are not accurate enough in determining the potential virulent strains. The impact of this study is that we have developed a subtyping approach that actually targets a virulence property. PMID- 12631207 TI - Production and molecular characteristics of four groups of exopolysaccharides from submerged culture of Phellinus gilvus. AB - AIMS: The objective of the present study was to determine the optimal culture conditions for the production of four groups of exopolysaccharides (EPSs) in Phellinus gilvus by submerged culture and to investigate their molecular properties by multi-angle laser-light scattering (MALLS) analysis. METHODS AND RESULTS: The optimal temperature and initial pH for the production of both mycelial biomass and EPSs in P. gilvus by submerged flask cultures were found to be 30 degrees C and pH 9.0, respectively. Glucose and corn steep powder were the most suitable carbon and nitrogen source for both mycelial biomass and EPS production. Optimal medium composition was determined to be glucose 30 g l-1, corn steep powder 5 g l-1, MgSO4 1.23 g l-1, KH2PO4 0.68 g l-1, and K2HPO4 0.87 g l-1. Four groups of EPSs (Fr-I, II, III, and IV) were obtained from the culture filtrates by gel filtration chromatography on Sepharose CL-4B and characterized by size exclusion chromatography (SEC) coupled with MALLS. The weight average molar mass (Mw) of Fr-I, Fr-II, Fr-III and Fr-IV were determined to be 8.628 x 106 (+/-129 420), 1.045 x 106 (+/-19 855), 61.09 x 104 (+/-1244), and 33.55 x 104 (+/-134) g mol-1, respectively. CONCLUSIONS: Under optimal culture conditions, the maximum EPS production in a 5-l stirred fermenter indicated 5.3 g l-1 after 11 days of fermentation. The SEC/MALLS analysis revealed that Fr-I, which has extremely high molecular weight, was presumably an aggregate of complex polysaccharides forming a compact globular shape; whereas Fr-II was nearly spherical, Fr-III and Fr-IV were rod-like chains in an aqueous solution. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on the production of high amounts of EPSs from liquid-culture of the basidiomycete, P. gilvus. The SEC/MALLS approach used in this study could be useful in providing greater insight into the characterization of the mushroom polysaccharides without carrying out elaborate fractionation procedures prior to analysis. PMID- 12631209 TI - Microbiological examination of ready-to-eat stuffing from retail premises in the north-east of England. The 'Get Stuffed' survey. AB - AIMS: To establish the microbiological quality of ready-to-eat stuffing from retail premises in the north-east of England. To establish threshold levels of bacteria in the product for acceptance as a ready-to-eat food. To determine the relationship between the microbiology of the product and production processes. METHODS AND RESULTS: A microbiological study of ready-to-eat stuffing using validated methods was performed on 147 samples from 139 retail premises. The determinants investigated were as follows: aerobic colony count, Escherichia coli, Enterobacteriaceae, Staphylococcus aureus, Bacillus spp., Salmonella spp. and Campylobacter spp. Results indicate that using current guidelines 76.3% were satisfactory, 15.6% were acceptable and 8.2% were of unsatisfactory quality. CONCLUSIONS: Unsatisfactory results were due to high aerobic colony counts, E. coli, Enterobacteriaceae and S. aureus. There were significant associations between bacteriological quality and temperature of storage, food hygiene training, product discard policy and confidence in management scores. SIGNIFICANCE AND IMPACT OF THE STUDY: The microbiology of ready-to-eat stuffing suggests that this is a relatively safe product. It is suggested that the product be placed in food category 3 in the current guidelines for ready-to-eat foods. PMID- 12631210 TI - Molecular characterization of Oenococcus oeni genes encoding proteins involved in arginine transport. AB - AIMS: This work was carried out to complete the sequence of the arc cluster involved in arginine catabolism in Oenococcus oeni, and particularly to characterize the genes encoding proteins involved in arginine transport. METHODS AND RESULTS: Using molecular cloning, two loci encoding proteins involved in the arginine-ornithine antiport were isolated. Their expression patterns were monitored by RT-PCR to study the influence of arginine on their transcription. Polycistronic mRNAs were detected. PCR performed directly on colonies with primer pairs specific of arc genes was used to discriminate strains able/unable to degrade arginine. CONCLUSIONS: Oenococcus oeni contains two arcD loci encoding similar proteins. Their expression is not influenced by arginine and polycistronic messengers were detected. The inability to use arginine is due to a lack of genetic information encoding proteins of the arginine deiminase pathway. SIGNIFICANCE AND IMPACT OF THE STUDY: The constitutive expression of arcD genes points to the positive role of arginine on O. oeni cell growth. The occasional presence of all the arc ABCD genes together in O. oeni strains might provide insights into the growth rate variability within this species. PMID- 12631211 TI - Theory of antimicrobial combinations: biocide mixtures - synergy or addition? AB - AIMS: To demonstrate the effect that non-linear dose responses have on the appearance of synergy in mixtures of antimicrobials. METHODS AND RESULTS: A mathematical model, which allows the prediction of the efficacy of mixtures of antimicrobials with non-linear dose responses, was produced. The efficacy of antimicrobial mixtures that would be classified as synergistic by time-kill methodology was shown to be a natural consequence of combining antimicrobials with non-linear dose responses. CONCLUSIONS: The effectiveness of admixtures of biocides and other antimicrobials with non-linear dose responses can be predicted. If the dose response (or dilution coefficient) of any biocidal component, in a mixture, is other than one, then the time-kill methodology used to ascertain the existence of synergy in antimicrobial combinations is flawed. SIGNIFICANCE AND IMPACT OF THE STUDY: The kinetic model developed allows the prediction of the efficacy of antimicrobial combinations. Combinations of known antimicrobials, which reduce the time taken to achieve a specified level of microbial inactivation, can be easily assessed once the kinetic profile of each component has been obtained. Most patented cases of antimicrobial synergy have not taken into account the possible effect of non-linear dose responses of the component materials. That much of the earlier literature can now be predicted, suggests that future cases will require more thorough proof of the alleged synergy. PMID- 12631212 TI - Pathologic prognostic factors in early stage adult-type granulosa cell tumors of the ovary. PMID- 12631213 TI - Health-related quality of life in gynecological oncology: instruments and psychometric properties. AB - Quality of life is generally recognized as a subjective, multidimensional concept, which places emphasis on the self-perception and subjective experience of the patient compared to the expectation of an individual's current health state. Health-related quality of life, which encompasses the psychological, physical, and social functioning of patients, has evolved over recent decades into an established treatment outcome in cancer clinical trials. Assessing quality of life as a clinical trial outcome enables clinicians to better address concerns of gynecological oncology patients, but selection of appropriate measurement tools is critical. This article reviews the concepts of reliability and validity, and describes three health-related quality of life instruments, their psychometric properties, and their use in gynecological oncology clinical trials and other cancer research. PMID- 12631214 TI - A decision aid for women at increased risk for ovarian cancer. AB - This paper reviews changes that have occurred within and without the medical profession that have fostered an increasing demand for decision aids as adjuncts to practitioners' counseling to prepare patients for decision making. In the absence of data on the efficacy of ovarian cancer screening and prophylactic strategies, decisions about optimal care are difficult for both women and their doctors. Because surveillance and preventive options are an area of great uncertainty, a decision aid has been developed specifically aimed at facilitating decisions involving ovarian cancer risk management options. This was achieved by reviewing and integrating the available literature on models of medical decision making, patient preferences for information and involvement in decision making, the utility of decision aids, and management options for ovarian cancer risk. Findings indicate that patients wish to be informed participants in the decision making process and that decision aids are an acceptable and effective method of providing quality information in a format that facilitates an inclusive model of shared decision making. A decision aid designed for women at increased risk of ovarian cancer that facilitates informed decision making may be a valuable addition to patient support. A randomized controlled trial of this type of educational material will provide timely and much needed evidence on its acceptability and efficacy. PMID- 12631215 TI - The possible association between in vitro fertilization treatments and cancer development. AB - The objective of this paper is to assess whether ovarian hyperstimulation and in vitro fertilization (IVF) are associated with increased risk of cancer development, using an historical cohort analysis of infertile women who attended the IVF unit, Lis Maternity Hospital Tel Aviv Medical Center, Tel Aviv, Israel. One thousand and 82 women participated in the IVF treatment program between 1984 and 1992. Cancer incidence rates were determined through the National Cancer Registry and were compared to the expected rates with respect to appropriate age and continent of birth. Twenty-one cases of cancer were observed as compared to 11 that were expected (SIR 1.91; 95% CI 1.18-2.91). When cancer cases that were diagnosed within one year of the IVF treatment were excluded from the analysis (SIR = 1.46; 95% CI 0.83-2.36), no significant excess risk of cancer was noted. We conclude that in this cohort of infertile women, the higher than expected cancer rate could not be attributed to IVF treatments. Special attention should be made to women who may be diagnosed with cancer during or shortly after IVF treatment. PMID- 12631216 TI - HER-2/neu expression in ovarian clear cell carcinomas. AB - HER-2 /neu is a 185-kDa glycoprotein and a transmembrane receptor with tyrosine kinase activity. Its overexpression is observed in 25-30% of primary breast carcinomas and is associated with a poor clinical prognosis. Recently, the U.S. Food and Drug Administration and the Japanese Ministry of Health, Welfare, and Labor approved the use of trastuzumab (Herceptin, Genentech, South San Francisco, CA) for the treatment of patients with metastatic breast carcinomas overexpressing HER-2 /neu. Results of clinical trials with Herceptin suggest that it may prolong the survival of patients with advanced metastatic breast carcinoma. Relatively little is known concerning the relationship between HER-2 /neu status and ovarian clear cell carcinoma. If HER-2 /neu overexpression status were demonstrable in ovarian clear cell carcinoma and a clinical correlation between overexpression and prognosis could be established, a rationale for clinical use of Herceptin for this tumor could be established. Our aim was to evaluate HER-2 /neu status in ovarian clear cell carcinomas. Fifteen ovarian clear cell carcinoma cases were immunostained for HER-2 /neu using HercepTest (DAKO, Glostrup, Denmark). Overexpression of HER-2 /neu was detected in only one case. Unlike in breast carcinoma, HER-2 /neu overexpression appeared to be uncommon in ovarian clear cell carcinomas. Herceptin may thus target only a small proportion of ovarian clear cell carcinomas and be of limited clinical value for treatment of this carcinoma. PMID- 12631217 TI - Simultaneous carcinoma of the endometrium and ovary vs endometrial carcinoma with ovarian metastases: a clinical and immunohistochemical determination. AB - The aim of this study was to perform a clinical and immunohistochemical comparison between simultaneous independent tumors involving endometrium and ovary and metastatic endometrial tumors, and to try to find clinical and /or immunohistochemical parameters differentiating between these two entities. Sixteen cases of simultaneous independent primaries of endometrium and ovary, presenting the same histologic type, were compared with 12 cases of primary endometrial cancer, demonstrating ovarian metastases. The comparison related to patients' characteristics and immunohistochemical expression of estrogen and progesterone receptors (ER,PR), bcl-2, HER-2 /neu, p53, and cell proliferation marker Ki-67 in endometrial and ovarian tumors. The only clinical parameter differentiating significantly between the groups was the prevalence of familial cancer, being more frequent in the group of metastatic tumors (P = 0.03). Immunohistochemical analysis demonstrated the same immunostaining in endometrium and ovary for all immunohistochemical parameters in cases of metastatic endometrial cancer. Conversely, 62.5% of cases with simultaneous tumors of endometrium and ovary could be differentiated from metastatic tumors by distinct immunohistochemical expression of ER and PR in endometrial and ovarian tumors (P = 0.0006), and 31.3% of cases could be differentiated by distinct immunostaining for bcl-2 (P = 0.03). Immunohistochemical parameters HER-2 /neu, p53 and Ki-67 were not appropriate for the distinction between the two study groups. We conclude that the application of immunohistochemical analysis may play an important role in the differentiation between cases of simultaneous independent carcinomas of endometrium and ovary vs. cases of endometrial carcinoma with ovarian metastases. PMID- 12631218 TI - Low risk endometrial cancer: a study of pelvic lymph node metastasis. AB - The aim of our study was to find preoperative or intraoperative pathologic indicators that would discriminate a subgroup of early corpus cancers that would not require lymphadenectomy. A retrospective review of the medical records of 107 patients with endometrioid adenocarcinoma, FIGO grade 1 or 2 tumor, myometrial invasion 80 mg L-1 regarding overall survival were 1.0, 3.3, and 0.7. Influence of lysozyme on survival was bimodal both in "non-monocytic" AML (HR 1.0, 3.0, and 0.1) and M4-M5 (HR 1.0, 10.1, and 1.2). Our finding of a bimodal relation between serum lysozyme and prognosis in AML should be regarded as a new hypothesis and controlled in other studies. PMID- 12631258 TI - CAL72: a human osteosarcoma cell line with unique effects on hematopoietic cells. AB - Permanent osteoblastic cell lines are potential tools to study the interactions between osteoblastic and hematopoietic cells in the bone marrow cavity. In a recent work we have shown that the osteosarcoma cell line CAL72 may be more closely related to normal osteoblasts than the osteosarcoma cells previously described. In the present work we continued the characterisation of the CAL72 cell line with regard to its effects on various hematopoietic cells, in coculture experiments. We show here that CAL72 cells, in contrast to MG-63 or SaOS-2 osteosarcoma cell lines, do not inhibit hematopoietic colony formation and sustain the limited expansion of hematopoietic progenitors in a similar way to that described for normal osteoblasts. We also demonstrate that CAL72 cells induce the monocytic differentiation of the promyelocytic HL-60 cell line like MG 63 and SaOS-2, but support a better maturation and a longer survival of the differentiated cells than the two other osteosarcoma cell lines. In order to better understand the differential effects observed between CAL72 and MG-63 or SaOS-2, we analysed the cytokine and chemokine mRNA expression of these cells using the RNase protection quantitative assay. We show here that the expression profile of CAL72 is clearly different from that of MG-63 or SaOS-2 and may explain, at least in part, its specific effects on hematopoietic cells. Taken together these experiments confirm that CAL72 has particular properties and is an interesting tool to study the role of osteoblastic cells in hematopoietic cell growth and differentiation. PMID- 12631259 TI - Surface antigen expression and correlation with variable heavy-chain gene mutation status in chronic lymphocytic leukemia. AB - Recent studies have demonstrated that B-cell chronic lymphocytic leukemia (CLL) consists of two clinical entities with either somatically hypermutated (M-CLL) or unmutated (UM-CLL) immunoglobulin variable heavy-chain (VH) regions. In view of the fact that the cellular biology of these two subsets of disease is currently unexplored, we performed an extensive analysis of the surface antigen expression and correlated this with the VH gene mutation status in a cohort of 32 CLL patients. Using polymerase chain reaction amplification and nucleotide sequencing, the VH genes were shown to be mutated in 10 cases (31%) and unmutated in 22 (69%). The expression of 27 surface membrane antigens in peripheral blood leukemic cells was analyzed by flow cytometry, measuring both the percentage of positive cells as well as the geometric mean fluorescence intensity (GMF). Most of the surface membrane antigens (CD5, CD11c, CD19, CD20, CD21, CD22, CD23, CD25, CD40, CD45, VD79b, CD80, CD95, CD122, CD124, CD126, CD130, CD154, IgM, and IgD) showed a similar expression pattern in both UM-CLL and M-CLL patients. The similarity of M-CLL and UM-CLL, as demonstrated here for the first time with many protein markers, indicates a considerably homogeneous phenotype in both subsets. Furthermore, CD27 was strongly expressed in all cases, which may suggest a memory cell phenotype for both M-CLL and UM-CLL. More positive cells in the UM-CLL group were observed regarding CD38, but CD38 was not a good predictor of VH gene mutation status. Seventy percent of the M-CLL cases, but only 36% of UM-CLL cases, were Ig-lambda+. The most striking differential expression, however, was observed in the two slicing variants of the common leukocyte antigen CD45, namely CD45RO and CD45RA. CD45RO expression was significantly associated with M-CLL, whereas the GMF intensity of CD45RA tended to be associated with UM-CLL. The role of these CD45 splicing variants in the pathogenesis of CLL deserves further investigation. PMID- 12631257 TI - Pamidronate is superior to ibandronate in decreasing bone resorption, interleukin 6 and beta 2-microglobulin in multiple myeloma. AB - OBJECTIVES: Bisphosphonates have been found to reduce skeletal events in patients with multiple myeloma (MM). This is the first randomised trial to compare the efficacy of pamidronate and ibandronate, a third-generation aminobisphosphonate, in bone turnover and disease activity in MM patients. METHODS: Patients with MM, stage II or III, were randomly assigned to receive either pamidronate 90 mg (group I: 23 patients) or ibandronate 4 mg (group II: 21 patients) as a monthly intravenous infusion in addition to conventional chemotherapy. Skeletal events, such as pathologic fractures, hypercalcaemia, and bone radiotherapy were analysed. Bone resorption markers [N-terminal cross-linking telopeptide of type-I collagen (NTX) and tartrate-resistant acid phosphatase type 5b (TRACP-5b)], bone formation markers (bone alkaline phosphatase and osteocalcin), markers of disease activity (paraprotein, CRP, beta 2-microglobulin), and interleukin-6 (IL-6) were also studied. RESULTS: In both groups, the combination of chemotherapy with either pamidronate or ibandronate produced a reduction in bone resorption and tumour burden as measured by NTX, IL-6, paraprotein, CRP, and beta 2 microglobulin from the second month of treatment, having no effect on bone formation. TRACP-5b also had a significant reduction in the pamidronate group from the second month of treatment and in the ibandronate group from the sixth month. However, there was a greater reduction of NTX, IL-6, and beta 2 microglobulin in group I than in group II, starting at the second month of treatment (P = 0.002, 0.001, and 0.004, respectively) and of TRACP-5b, starting at the fourth month (P = 0.014), that being continued throughout the 10-month follow-up of this study. There was no difference in skeletal events during this period. A significant correlation was observed between changes of NTX and changes of TRACP-5b, IL-6, and beta 2-microglobulin from the second month for patients of both groups. CONCLUSIONS: These results suggest that a monthly dose of 90 mg of pamidronate is more effective than 4 mg of ibandronate in reducing osteoclast activity, bone resorption, IL-6, and possibly tumour burden in MM. TRACP-5b has also proved to be a useful new marker for monitoring bisphosphonates treatment in MM. PMID- 12631260 TI - Severe IgA-mediated auto-immune haemolytic anaemia in a 48-yr-old woman. AB - Auto-immune haemolytic anaemia (AIHA) is characterised by haemolysis associated with the presence of the immunoglobulins IgG, IgM or IgA, and/or components of the complement system on the red cell membrane. The immunoglobulins react as auto antibodies against the red cell antigens of the patient. IgG antibodies and the complement component C3d can be detected by the direct antiglobulin test (DAT); however, IgM and particularly IgA antibodies may not necessarily be detected by the broad-spectrum anti-human-globulin serum. We present the case of a 48-yr-old woman with severe AIHA. The initial polyspecific direct antiglobulin test (DAT), using a broad-spectrum antiserum, was negative. Testing with monospecific antisera led to the diagnosis of AIHA due to warm-acting auto-antibodies solely of the IgA class, which is a very rare finding. As therapy with steroids alone did not lead to a lasting remission, splenectomy was performed 10 months after initial diagnosis. There has been no relapse of AIHA since, even after steroid medication was withdrawn and even though the monospecific IgA-DAT has remained positive. This case demonstrates the importance of performing a monospecific antiglobulin test if there is a strong suspicion of AIHA in apparently "Coombs negative" haemolytic anaemia. In AIHA caused by solely IgA antibodies, the polyspecific direct antiglobulin test may be negative or only weakly positive because of a limited content of anti-IgA antibodies in the polyspecific anti human-globulin serum. First-line treatment of warm-type AIHA is the administration of high-dose glucocorticosteroids; splenectomy is indicated in steroid-refractory patients. PMID- 12631261 TI - Unstable angina associated with coronary arterial calcification in a thalassemia intermedia patient with a pseudoxanthoma elasticum-like syndrome. AB - The coexistence of a pseudoxanthoma elasticum (PXE)-like syndrome in beta thalassemia and other hemoglobinopathies is a recently established clinical entity that has been observed with a significant frequency and related to some severe, even life-threatening complications. We present here a thalassemia intermedia patient who developed unstable angina in a setting of severe anemia and PXE-related coronary arterial calcification. Besides the clinical significance of this PXE-like syndrome, its acquired nature may introduce some new thoughts regarding the pathogenesis of atherosclerosis. PMID- 12631262 TI - 2-Oxo acid dehydrogenase complexes of multicellular organisms. PMID- 12631263 TI - 2-Oxo acid dehydrogenase complexes in redox regulation. AB - A number of cellular systems cooperate in redox regulation, providing metabolic responses according to changes in the oxidation (or reduction) of the redox active components of a cell. Key systems of central metabolism, such as the 2-oxo acid dehydrogenase complexes, are important participants in redox regulation, because their function is controlled by the NADH/NAD+ ratio and the complex-bound dihydrolipoate/lipoate ratio. Redox state of the complex-bound lipoate is an indicator of the availability of the reaction substrates (2-oxo acid, CoA and NAD+) and thiol-disulfide status of the medium. Accumulation of the dihydrolipoate intermediate causes inactivation of the first enzyme of the complexes. With the mammalian pyruvate dehydrogenase, the phosphorylation system is involved in the lipoate-dependent regulation, whereas mammalian 2-oxoglutarate dehydrogenase exhibits a higher sensitivity to direct regulation by the complex bound dihydrolipoate/lipoate and external SH/S-S, including mitochondrial thioredoxin. Thioredoxin efficiently protects the complexes from self inactivation during catalysis at low NAD+. As a result, 2-oxoglutarate dehydrogenase complex may provide succinyl-CoA for phosphorylation of GDP and ADP under conditions of restricted NAD+ availability. This may be essential upon accumulation of NADH and exhaustion of the pyridine nucleotide pool. Concomitantly, thioredoxin stimulates the complex-bound dihydrolipoate-dependent production of reactive oxygen species. It is suggested that this side-effect of the 2-oxo acid oxidation at low NAD+in vivo would be overcome by cooperation of mitochondrial thioredoxin and the thioredoxin-dependent peroxidase, SP-22. PMID- 12631264 TI - Regulation of pyruvate dehydrogenase complex activity in plant cells. AB - The pyruvate dehydrogenase complex (PDC) is subjected to multiple interacting levels of control in plant cells. The first level is subcellular compartmentation. Plant cells are unique in having two distinct, spatially separated forms of the PDC; mitochondrial (mtPDC) and plastidial (plPDC). The mtPDC is the site of carbon entry into the tricarboxylic acid cycle, while the plPDC provides acetyl-CoA and NADH for de novo fatty acid biosynthesis. The second level of regulation of PDC activity is the control of gene expression. The genes encoding the subunits of the mt- and plPDCs are expressed following developmental programs, and are additionally subject to physiological and environmental cues. Thirdly, both the mt- and plPDCs are sensitive to product inhibition, and, potentially, to metabolite effectors. Finally, the two different forms of the complex are regulated by distinct organelle-specific mechanisms. Activity of the mtPDC is regulated by reversible phosphorylation catalyzed by intrinsic kinase and phosphatase components. An additional level of sensitivity is provided by metabolite control of the kinase activity. The plPDC is not regulated by reversible phosphorylation. Instead, activity is controlled to a large extent by the physical environment that exists in the plastid stroma. PMID- 12631266 TI - Plasmoredoxin, a novel redox-active protein unique for malarial parasites. AB - Thioredoxins are a group of small redox-active proteins involved in cellular redox regulatory processes as well as antioxidant defense. Thioredoxin, glutaredoxin, and tryparedoxin are members of the thioredoxin superfamily and share structural and functional characteristics. In the malarial parasite, Plasmodium falciparum, a functional thioredoxin and glutathione system have been demonstrated and are considered to be attractive targets for antimalarial drug development. Here we describe the identification and characterization of a novel 22 kDa redox-active protein in P. falciparum. As demonstrated by in silico sequence analyses, the protein, named plasmoredoxin (Plrx), is highly conserved but found exclusively in malarial parasites. It is a member of the thioredoxin superfamily but clusters separately from other members in a phylogenetic tree. We amplified the gene from a gametocyte cDNA library and overexpressed it in E. coli. The purified gene product can be reduced by glutathione but much faster by dithiols like thioredoxin, glutaredoxin, trypanothione and tryparedoxin. Reduced Plrx is active in an insulin-reduction assay and reduces glutathione disulfide with a rate constant of 640 m-1.s-1 at pH 6.9 and 25 degrees C; glutathione dependent reduction of H2O2 and hydroxyethyl disulfide by Plrx is negligible. Furthermore, plasmoredoxin provides electrons for ribonucleotide reductase, the enzyme catalyzing the first step of DNA synthesis. As demonstrated by Western blotting, the protein is present in blood-stage forms of malarial parasites. Based on these results, plasmoredoxin offers the opportunity to improve diagnostic tools based on PCR or immunological reactions. It may also represent a specific target for antimalarial drug development and is of phylogenetic interest. PMID- 12631265 TI - Essential roles of lipoyl domains in the activated function and control of pyruvate dehydrogenase kinases and phosphatase isoform 1. AB - Four pyruvate dehydrogenase kinase and two pyruvate dehydrogenase phosphatase isoforms function in adjusting the activation state of the pyruvate dehydrogenase complex (PDC) through determining the fraction of active (nonphosphorylated) pyruvate dehydrogenase component. Necessary adaptations of PDC activity with varying metabolic requirements in different tissues and cell types are met by the selective expression and pronounced variation in the inherent functional properties and effector sensitivities of these regulatory enzymes. This review emphasizes how the foremost changes in the kinase and phosphatase activities issue from the dynamic, effector-modified interactions of these regulatory enzymes with the flexibly held outer domains of the core-forming dihydrolipoyl acetyl transferase component. PMID- 12631267 TI - Modeled ligand-protein complexes elucidate the origin of substrate specificity and provide insight into catalytic mechanisms of phenylalanine hydroxylase and tyrosine hydroxylase. AB - NMR spectroscopy and X-ray crystallography have provided important insight into structural features of phenylalanine hydroxylase (PAH) and tyrosine hydroxylase (TH). Nevertheless, significant problems such as the substrate specificity of PAH and the different susceptibility of TH to feedback inhibition by l-3,4 dihydroxyphenylalanine (l-DOPA) compared with dopamine (DA) remain unresolved. Based on the crystal structures 5pah for PAH and 2toh for TH (Protein Data Bank), we have used molecular docking to model the binding of 6(R)-l-erythro-5,6,7,8 tetrahydrobiopterin (BH4) and the substrates phenylalanine and tyrosine to the catalytic domains of PAH and TH. The amino acid substrates were placed in positions common to both enzymes. The productive position of tyrosine in TH.BH4 was stabilized by a hydrogen bond with BH4. Despite favorable energy scores, tyrosine in a position trans to PAH residue His290 or TH residue His336 interferes with the access of the essential cofactor dioxygen to the catalytic center, thereby blocking the enzymatic reaction. DA and l-DOPA were directly coordinated to the active site iron via the hydroxyl residues of their catechol groups. Two alternative conformations, rotated 180 degrees around an imaginary iron-catecholamine axis, were found for DA and l-DOPA in PAH and for DA in TH. Electrostatic forces play a key role in hindering the bidentate binding of the immediate reaction product l-DOPA to TH, thereby saving the enzyme from direct feedback inhibition. PMID- 12631268 TI - Human acid sphingomyelinase. AB - Human acid sphingomyelinase (haSMase, EC 3.1.4.12) catalyzes the lysosomal degradation of sphingomyelin to ceramide and phosphorylcholine. An inherited haSMase deficiency leads to Niemann-Pick disease, a severe sphingolipid storage disorder. The enzyme was purified and cloned over 10 years ago. Since then, only a few structural properties of haSMase have been elucidated. For understanding of its complex functions including its role in certain signaling and apoptosis events, complete structural information about the enzyme is necessary. Here, the identification of the disulfide bond pattern of haSMase is reported for the first time. Functional recombinant enzyme expressed in SF21 cells using the baculovirus expression system was purified and digested by trypsin. MALDI-MS analysis of the resulting peptides revealed the four disulfide bonds Cys120-Cys131, Cys385 Cys431, Cys584-Cys588 and Cys594-Cys607. Two additional disulfide bonds (Cys221 Cys226 and Cys227-Cys250) which were not directly accessible by tryptic cleavage, were identified by a combination of a method of partial reduction and MALDI-PSD analysis. In the sphingolipid activator protein (SAP)-homologous N-terminal domain of haSMase, one disulfide bond was assigned as Cys120-Cys131. The existence of two additional disulfide bridges in this region was proved, as was expected for the known disulfide bond pattern of SAP-type domains. These results support the hypothesis that haSMase possesses an intramolecular SAP-type activator domain as predicted by sequence comparison [Ponting, C.P. (1994) Protein Sci., 3, 359-361]. An additional analysis of haSMase isolated from human placenta shows that the recombinant and the native human protein possess an identical disulfide structure. PMID- 12631269 TI - Enhanced expression of Mcm proteins in cancer cells derived from uterine cervix. AB - Minichromosome maintenance proteins (Mcm) 2-7 play essential roles in eukaryotic DNA replication. Several reports have indicated the usefulness of Mcm proteins as markers of cancer cells in histopathological diagnosis. However, their mode of expression and pathophysiological significance in cancer cells remain to be clarified. We compared the level of expression of Mcm proteins among human HeLa uterine cervical carcinoma cells, SV40-transformed human fibroblast GM00637 cells and normal human fibroblast WI-38 cells. All the proteins examined were detected in HeLa and GM cells at 6-10 times the level found in WI-38 cells on average. This increase was observed both in total cellular proteins and in the chromatin bound fraction. Consistently, Mcm2 mRNA was enriched in HeLa cells to approximately four times the level in WI-38 cells, and the synthesis of Mcm4, 6 and 7 proteins was accelerated in HeLa cells. Immunohistochemical studies of surgical materials from human uterine cervix showed that Mcm3 and 4 are ubiquitously expressed in cancer cells. Further, the positive rate and level of Mcm3 and 4 expression appeared to be higher in cancer cells than in normal proliferating cells of the uterine cervix and dysplastic cells, suggesting that they can be useful markers to distinguish these cells. PMID- 12631270 TI - Outer sphere mutagenesis of Lactobacillus plantarum manganese catalase disrupts the cluster core. Mechanistic implications. AB - X-ray crystallography of the nonheme manganese catalase from Lactobacillus plantarum (LPC) [Barynin, V.V., Whittaker, M.M., Antonyuk, S.V., Lamzin, V.S., Harrison, P.M., Artymiuk, P.J. & Whittaker, J.W. (2001) Structure9, 725-738] has revealed the structure of the dimanganese redox cluster together with its protein environment. The oxidized [Mn(III)Mn(III)] cluster is bridged by two solvent molecules (oxo and hydroxo, respectively) together with a micro 1,3 bridging glutamate carboxylate and is embedded in a web of hydrogen bonds involving an outer sphere tyrosine residue (Tyr42). A novel homologous expression system has been developed for production of active recombinant LPC and Tyr42 has been replaced by phenylalanine using site-directed mutagenesis. Spectroscopic and structural studies indicate that disruption of the hydrogen-bonded web significantly perturbs the active site in Y42F LPC, breaking one of the solvent bridges and generating an 'open' form of the dimanganese cluster. Two of the metal ligands adopt alternate conformations in the crystal structure, both conformers having a broken solvent bridge in the dimanganese core. The oxidized Y42F LPC exhibits strong optical absorption characteristic of high spin Mn(III) in low symmetry and lower coordination number. MCD and EPR measurements provide complementary information defining a ferromagnetically coupled electronic ground state for a cluster containing a single solvent bridge, in contrast to the diamagnetic ground state found for the native cluster containing a pair of solvent bridges. Y42F LPC has less than 5% of the catalase activity and much higher Km for H2O2 ( approximately 1.4 m) at neutral pH than WT LPC, although the activity is slightly restored at high pH where the cluster is converted to a diamagnetic form. These studies provide new insight into the contribution of the outer sphere tyrosine to the stability of the dimanganese cluster and the role of the solvent bridges in catalysis by dimanganese catalases. PMID- 12631271 TI - Transport of taurocholate by mutants of negatively charged amino acids, cysteines, and threonines of the rat liver sodium-dependent taurocholate cotransporting polypeptide Ntcp. AB - The relevance of functional amino acids for taurocholate transport by the sodium dependent taurocholate cotransporting polypeptide Ntcp was determined by site directed mutagenesis. cRNA from 28 single-points mutants of the rat liver Ntcp clone was expressed in Xenopus laevis oocytes. Mutations were generated in five conserved negatively charged amino acids (aspartates and glutamates) which were present in nine members of the SBAT-family, in two nonconserved negatively charged amino acids, in all eight Ntcp-cysteines, and in two threonines from a protein kinase C consensus region of the Ntcp C-terminus. Functional amino acids were Asp115, Glu257, and Cys266, which were found to be essential for the maintenance of taurocholic acid transport. Asp115 is located in the large intracellular loop III, whereas Glu257 and Cys266 are located in the large extracellular loop VI. Four mutations of threonines from the C-terminus of the Ntcp by alanines or tyrosines showed no effects on sodium-dependent taurocholate transport. Introduction of the FLAG(R) motif into several transport negative point mutations demonstrated that all mutated proteins besides one were present within the cell membrane of the oocytes and provided proof that an insertion defect has not caused transport deficiency by these Ntcp mutants. The latter was observed only with the transport negative mutant Asp24Asn. In conclusion, loop amino acids are required for sodium-dependent substrate translocation by the Ntcp. PMID- 12631272 TI - Transcription termination at the mouse mitochondrial H-strand promoter distal site requires an A/T rich sequence motif and sequence specific DNA binding proteins. AB - Termination of mitochondrial (mt) H-strand transcription in mammalian cells occurs at two distinct sites on the genome. The first site of termination, referred to as mt-TERM occurs beyond the 16 S rRNA gene. However, the second and final site of termination beyond the tRNAThr gene remains unclear. In this study we have characterized the site of termination of the polycistronic distal gene transcript beyond the D-loop region, immediately upstream of the tRNAPhe gene. This region, termed D-TERM, maps to nucleotides 16274-16295 of the mouse genome and includes a conserved A/T rich sequence motif AATAAA as a part of the terminator. Gel-shift analysis showed that the 22 bp D-TERM DNA forms two major complexes with mouse liver mt extract in a sequence-specific manner. Protein purification by DNA-affinity chromatography yielded two major proteins of 45 kDa and 70 kDa. Finally, the D-TERM DNA can mediate transcription termination in a unidirectional manner in a HeLa mt transcription system, only in the presence of purified mouse liver mt D-TERM DNA binding proteins. We have therefore characterized a novel mt transcription termination system, similar in some properties to that of sea urchin, as well as the nuclear RNA Pol I and Pol II transcription termination systems. PMID- 12631273 TI - nNOS inhibition, antimicrobial and anticancer activity of the amphibian skin peptide, citropin 1.1 and synthetic modifications. The solution structure of a modified citropin 1.1. AB - A large number of bioactive peptides have been isolated from amphibian skin secretions. These peptides have a variety of actions including antibiotic and anticancer activities and the inhibition of neuronal nitric oxide synthase. We have investigated the structure-activity relationship of citropin 1.1, a broad spectrum antibiotic and anticancer agent that also causes inhibition of neuronal nitric oxide synthase, by making a number of synthetically modified analogues. Citropin 1.1 has been shown previously to form an amphipathic alpha-helix in aqueous trifluoroethanol. The results of the structure-activity studies indicate the terminal residues are important for bacterial activity and increasing the overall positive charge, while maintaining an amphipathic distribution of residues, increases activity against Gram-negative organisms. Anticancer activity generally mirrors antibiotic activity suggesting a common mechanism of action. The N-terminal residues are important for inhibition of neuronal nitric oxide synthase, as is an overall positive charge greater than three. The structure of one of the more active synthetic modifications (A4K14-citropin 1.1) was determined in aqueous trifluoroethanol, showing that this peptide also forms an amphipathic alpha-helix. PMID- 12631274 TI - Identification of sites of phosphorylation by G-protein-coupled receptor kinase 2 in beta-tubulin. AB - G-protein-coupled receptor kinase 2 (GRK2) is known to specifically phosphorylate the agonist-bound forms of G-protein-coupled receptors (GPCRs). This strict specificity is due at least partly to activation of GRK2 by agonist-bound GPCRs, in which basic residues in intracellular regions adjacent to transmembrane segments are thought to be involved. Tubulin was found to be phosphorylated by GRK2, but it remains unknown if tubulin can also serve as both a substrate and an activator for GRK2. Purified tubulin, phosphorylated by GRK2, was subjected to biochemical analysis, and the phosphorylation sites in beta-tubulin were determined to be Thr409 and Ser420. In addition, the Ser444 in beta III-tubulin was also indicated to be phosphorylated by GRK2. The phosphorylation sites in tubulin for GRK2 reside in the C-terminal domain of beta-tubulin, which is on the outer surface of microtubules. Pretreatment of tubulin with protein phosphatase type-2A (PP2A) resulted in a twofold increase in the phosphorylation of tubulin by GRK2. These results suggest that tubulin is phosphorylated in situ probably by GRK2 and that the phosphorylation may affect the interaction of microtubules with microtubule-associated proteins. A GST fusion protein of a C-terminal region of beta I-tubulin (393-445 residues), containing 19 acidic residues but only one basic residue, was found to be a good substrate for GRK2, like full-length beta tubulin. These results, together with the finding that GRK2 may phosphorylate synuclein and phosducin in their acidic domains, indicate that some proteins with very acidic regions but without basic activation domains could serve as substrates for GRK2. PMID- 12631275 TI - Determination of the redox potentials and electron transfer properties of the FAD and FMN-binding domains of the human oxidoreductase NR1. AB - Human novel reductase 1 (NR1) is an NADPH dependent diflavin oxidoreductase related to cytochrome P450 reductase (CPR). The FAD/NADPH- and FMN-binding domains of NR1 have been expressed and purified and their redox properties studied by stopped-flow and steady-state kinetic methods, and by potentiometry. The midpoint reduction potentials of the oxidized/semiquinone (-315 +/- 5 mV) and semiquinone/dihydroquinone (-365 +/- 15 mV) couples of the FAD/NADPH domain are similar to those for the FAD/NADPH domain of human CPR, but the rate of hydride transfer from NADPH to the FAD/NADPH domain of NR1 is approximately 200-fold slower. Hydride transfer is rate-limiting in steady-state reactions of the FAD/NADPH domain with artificial redox acceptors. Stopped-flow studies indicate that hydride transfer from the FAD/NADPH domain of NR1 to NADP+ is faster than hydride transfer in the physiological direction (NADPH to FAD), consistent with the measured reduction potentials of the FAD couples [midpoint potential for FAD redox couples is -340 mV, cf-320 mV for NAD(P)H]. The midpoint reduction potentials for the flavin couples in the FMN domain are -146 +/- 5 mV (oxidized/semiquinone) and -305 +/- 5 mV (semiquinone/dihydroquinone). The FMN oxidized/semiquinone couple indicates stabilization of the FMN semiquinone, consistent with (a) a need to transfer electrons from the FAD/NADPH domain to the FMN domain, and (b) the thermodynamic properties of the FMN domain in CPR and nitric oxide synthase. Despite overall structural resemblance of NR1 and CPR, our studies reveal thermodynamic similarities but major kinetic differences in the electron transfer reactions catalysed by the flavin-binding domains. PMID- 12631276 TI - Lipid droplet and milk lipid globule membrane associated placental protein 17b (PP17b) is involved in apoptotic and differentiation processes of human epithelial cervical carcinoma cells. AB - The intracellular role of placental protein 17b (PP17b)/TIP47 has been controversial, because it is considered to be a protein required for mannose 6 phosphate receptor transport from endosome to trans-Golgi as well as a neutral lipid droplet-associated protein. The similarity between the amino acid sequences of PP17 variants, adipophilin and perilipins, and between their gene structures indicate that PP17b as well as other alternatively spliced PP17 variants belong to the lipid storage droplet protein family, containing also some differentiation factors. Using a specific antibody, PP17b was detected in lipid droplet fractions and co-localized with neutral lipid droplets stained by Nile red, and fluorescently labelled PP17 antibody in HeLa cells with confocal microscopy. PP17b was also detected in milk, associated to milk lipid globule membranes. Cytostatic agents induced apoptosis and PP17b synthesis in HeLa cells, which was significantly inhibited by protein kinase C (PKC) inhibitor, indicating the involvement of NF-kappa B and AP-1 transcription factors in this process, while protein kinase A (PKA) inhibitor had only a modest inhibitory effect. Cell differentiation induced by dibutyryl cyclic AMP or phorbol myristate acetate also increased PP17b synthesis, demonstrating its strong involvement in cell differentiation. PP17b synthesis was higher in M than in G0/G1 phases in control, apoptotic and differentiated cells. This data shows that PP17b is a neutral lipid droplet-associated protein, and its expression is regulated by PKC- and PKA dependent pathways. PMID- 12631277 TI - Disruption of structural and functional integrity of alpha 2-macroglobulin by cathepsin E. AB - alpha 2-Macroglobulin (alpha 2M) is an abundant glycoprotein with the intrinsic capacity for capturing diverse proteins for rapid delivery into cells. After internalization by the receptor- mediated endocytosis, alpha 2M-protein complexes were rapidly degraded in the endolysosome system. Although this is an important pathway for clearance of both alpha 2M and biological targets, little is known about the nature of alpha 2M degradation in the endolysosome system. To investigate the possible involvement of intracellular aspartic proteinases in the disruption of structural and functional integrity of alpha 2M in the endolysosome system, we examined the capacity of alpha 2M for interacting with cathepsin E and cathepsin D under acidic conditions and the nature of its degradation. alpha 2M was efficiently associated with cathepsin E under acidic conditions to form noncovalent complexes and rapidly degraded through the generation of three major proteins with apparent molecular masses of 90, 85 and 30 kDa. Parallel with this reaction, alpha 2M resulted in the rapid loss of its antiproteolytic activity. Analysis of the N-terminal amino-acid sequences of these proteins revealed that alpha 2M was selectively cleaved at the Phe811-Leu812 bond in about 100mer downstream of the bait region. In contrast, little change was observed for alpha 2M treated by cathepsin D under the same conditions. Together, the synthetic SPAFLA peptide corresponding to the Ser808-Ala813 sequence of human alpha 2M, which contains the cathepsin E-cleavage site, was selectively cleaved by cathepsin E, but not cathepsin D. These results suggest the possible involvement of cathepsin E in disruption of the structural and functional integrity of alpha 2M in the endolysosome system. PMID- 12631278 TI - Interaction of ostreolysin, a cytolytic protein from the edible mushroom Pleurotus ostreatus, with lipid membranes and modulation by lysophospholipids. AB - Ostreolysin is a 16-kDa cytolytic protein specifically expressed in primordia and fruiting bodies of the edible mushroom Pleurotus ostreatus. To understand its interaction with lipid membranes, we compared its effects on mammalian cells, on vesicles prepared with either pure lipids or total lipid extracts, and on dispersions of lysophospholipids or fatty acids. At nanomolar concentrations, the protein lysed human, bovine and sheep erythrocytes by a colloid-osmotic mechanism, compatible with the formation of pores of 4 nm diameter, and was cytotoxic to mammalian tumor cells. A search for lipid inhibitors of hemolysis revealed a strong effect of lysophospholipids and fatty acids, occurring below their critical micellar concentration. This effect was distinct from the capacity of ostreolysin to bind to and permeabilize lipid membranes. In fact, permeabilization of vesicles occurred only when they were prepared with lipids extracted from erythrocytes, and not with lipids extracted from P. ostreatus or pure lipid mixtures, even if lysophospholipids or fatty acids were included. Interaction with lipid vesicles, and their permeabilization, correlated with an increase in the intrinsic fluorescence and alpha-helical content of the protein, and with aggregation, which were not detected with lysophospholipids. It appears that either an unknown lipid acceptor or a specific lipid complex is required for binding, aggregation and pore formation. The inhibitory effect of lysophospholipids may reflect a regulatory role for these components on the physiological action of ostreolysin and related proteins during fruiting. PMID- 12631279 TI - Membrane targeting of a folded and cofactor-containing protein. AB - Targeting of proteins to and translocation across the membranes is a fundamental biological process in all organisms. In bacteria, the twin arginine translocation (Tat) system can transport folded proteins. Here, we demonstrate in vivo that the high potential iron-sulfur protein (HiPIP) from Allochromatium vinosum is translocated into the periplasmic space by the Tat system of Escherichia coli. In vitro, reconstituted HiPIP precursor (preHoloHiPIP) was targeted to inverted membrane vesicles from E. coli by a process requiring ATP when the Tat substrate was properly folded. During membrane targeting, the protein retained its cofactor, indicating that it was targeted in a folded state. Membrane targeting did not require a twin arginine motif and known Tat system components. On the basis of these findings, we propose that a pathway exists for the insertion of folded cofactor-containing proteins such as HiPIP into the bacterial cytoplasmic membrane. PMID- 12631280 TI - Disease-related mutations in cytochrome c oxidase studied in yeast and bacterial models. AB - Mitochondrial cytochrome c oxidase is a key protonmotive component of the respiratory chain. Mutations in the mitochondrially-encoded subunits of the complex have been reported in association with a range of diseases. In this work we used yeast and bacterial mutants to assess the effect of human mutations in subunit 1 (L196I) and subunit 3 (G78S, A200T, Delta F94-F98, F251L and W249Stop). While the stop mutation at the C-terminus of subunit 3 and the short deletion were highly deleterious and abolished the assembly of the mitochondrial enzyme, the four missense mutations caused little or no effect on the respiratory function. Detailed analysis of G78S, A200T and Delta F94-F98 in Rhodobacter sphaeroides confirmed and extended these observations. We show in this study that the combination of yeast and bacterial models is a useful tool to elucidate the effect of mutations in the catalytic core of cytochrome oxidase. The yeast enzyme is highly similar to the human enzyme and provides a good model to assess the deleterious effect of reported mutations. The bacterial system allows detailed biochemical analysis of the effect of the mutations on the function and assembly of the catalytic core of the enzyme. PMID- 12631281 TI - Adaptability and flexibility of HIV-1 protease. AB - Even though more than 200 three-dimensional structures of HIV-1 protease complexed to a variety of inhibitors are available in the Protein Data Bank; very few structures of unliganded protein have been determined. We have recently solved structures of unliganded HIV-1 protease tethered dimer mutants to resolutions of 1.9 A and 2.1 A, and have found that the flaps assume closed-flap conformation even in the absence of any bound ligand. We report comparison of the unliganded closed-flap structure with structures of HIV-1 protease inhibitor complexes with a view to accurately identifying structural changes that the ligand can induce on binding to HIV-1 protease in the crystal. These studies reveal that the least flexible region present in the active site of HIV-1 protease need not also be the least adaptable to external stress, thus highlighting the conceptual difference between flexibility and adaptability of proteins in general. PMID- 12631282 TI - Interactions of the antimicrobial beta-peptide beta-17 with phospholipid vesicles differ from membrane interactions of magainins. AB - We have studied the interaction of beta-17, a potent synthetic antimicrobial beta peptide, with phospholipids. We find that unlike other antimicrobial peptides such as magainin II, beta-17 facilitates the formation of nonbilayer phases, indicating that the peptide promotes negative curvature. Studies of liposomal leakage also indicate a different mode of membrane interaction relative to magainin II, but both leakage and membrane binding show that beta-17, like magainin II, has strong affinity for membranes containing anionic lipids. This is likely to be an important factor contributing to the antimicrobial specificity of the beta-peptide. PMID- 12631283 TI - Reduction of S-nitrosoglutathione by human alcohol dehydrogenase 3 is an irreversible reaction as analysed by electrospray mass spectrometry. AB - Human alcohol dehydrogenase 3/glutathione-dependent formaldehyde dehydrogenase was shown to rapidly and irreversibly catalyse the reductive breakdown of S nitrosoglutathione. The steady-state kinetics of S-nitrosoglutathione reduction was studied for the wild-type and two mutated forms of human alcohol dehydrogenase 3, mutations that have previously been shown to affect the oxidative efficiency for the substrate S-hydroxymethylglutathione. Wild-type enzyme readily reduces S-nitrosoglutathione with a kcat/Km approximately twice the kcat/Km for S-hydroxymethylglutathione oxidation, resulting in the highest catalytic efficiency yet identified for a human alcohol dehydrogenase. In a similar manner as for S-hydroxymethylglutathione oxidation, the catalytic efficiency of S-nitrosoglutathione reduction was significantly decreased by replacement of Arg115 by Ser or Lys, supporting similar substrate binding. NADH was by far a better coenzyme than NADPH, something that previously has been suggested to prevent reductive reactions catalysed by alcohol dehydrogenases through the low cytolsolic NADH/NAD+ ratio. However, the major products of S nitrosoglutathione reduction were identified by electrospray tandem mass spectrometry as glutathione sulfinamide and oxidized glutathione neither of which, in their purified form, served as substrate or inhibitor for the enzyme. Hence, the reaction products are not substrates for alcohol dehydrogenase 3 and the overall reaction is therefore irreversible. We propose that alcohol dehydrogenase 3 catalysed S-nitrosoglutathione reduction is of physiological relevance in the metabolism of NO in humans. PMID- 12631284 TI - TRAF6 and C-SRC induce synergistic AP-1 activation via PI3-kinase-AKT-JNK pathway. AB - Interleukin-1 (IL-1) induces multiple genes via activation of transcription factors that include NF-kappa B and activator protein-1 (AP-1). We found that IL 1-mediated c-Src activation was required for AP-1 activation, but not for NF kappa B activation and also revealed that c-Src-induced AP-1 activation was enhanced synergistically by the coexpression of TNF receptor associated factor 6 (TRAF6). In addition, c-Src interacts with TRAF6 in response to IL-1 and this interaction is required for c-Src activity. However, neither dominant negative mutants of TRAF6 (TRAF6 DN) nor kinase-dead mutant of c-Src (c-Src KD) counteracted each-induced AP-1 activation, suggesting no hierarchy between these two molecules. During the TRAF6 and c-Src-induced AP-1 activation, phosphatidylinositol 3 (PI3)-kinase, its downstream signaling molecule, Akt and c Jun N-terminal kinase (JNK) were significantly activated and inhibition of these kinase activities down-regulated AP-1 activation through the suppression of c-fos expression. Furthermore, TRAF6 and c-Src-induced JNK activation was significantly inhibited by PI3-kinase inhibitor or a dominant negative mutant of Akt (Akt DN). Taken together, our results demonstrate that c-Src and TRAF6 are key mediators of IL-1-induced AP-1 activation and provide evidence of cross talk between c-Src and TRAF6 molecules through PI3 kinase-Akt-JNK pathways. PMID- 12631285 TI - A possible physiological function and the tertiary structure of a 4-kDa peptide in legumes. AB - Previously, we isolated a 4-kDa peptide capable of binding to a 43-kDa receptor like protein and stimulating protein kinase activity of the 43-kDa protein in soybean. Both of them were found to localize in the plasma membranes and cell walls. Here, we report the physiological effects of 4-kDa peptide expressed transiently in the cultured carrot and bird's-foot trefoil cells transfected with pBI 121 plasmid containing the 4-kDa peptide gene. At early developmental stage, the transgenic callus grew rapidly compared to the wild callus in both species. Cell proliferation of in vitro cultured nonembryogenic carrot callus was apparently affected with the 4-kDa peptide in the medium. Complementary DNAs encoding the 4-kDa peptide from mung bean and azuki bean were cloned by PCR and sequenced. The amino-acid sequences deduced from the nucleotide sequences are homologous among legume species, particularly, the sites of cysteine residues are highly conserved. This conserved sequence reflects the importance of intradisulfide bonds required for the 4-kDa peptide to perform its function. Three dimensional structure of the 4-kDa peptide determined by NMR spectroscopy suggests that this peptide is a T-knot scaffold containing three beta-strands, and the specific binding activity to the 43-kDa protein and stimulatory effect on the protein phosphorylation could be attributed to the spatial arrangements of hydrophobic residues at the solvent-exposed surface of two-stranded beta-sheet of 4-kDa peptide. The importance of these residues for the 4-kDa peptide to bind to the 43-kDa protein was indicated by site-directed mutagenesis. These results suggest that the 4-kDa peptide is a hormone-like peptide and the 43-kDa protein is involved in cellular signal transduction of the peptide. PMID- 12631287 TI - DNA-binding and transcription characteristics of three cloned sigma factors from mustard (Sinapis alba L.) suggest overlapping and distinct roles in plastid gene expression. AB - We have isolated and studied the cloned sigma factors SASIG1-3 from mustard (Sinapis alba). In functional analyses using both promoter and factor mutants, the three recombinant proteins all had similar basic properties but also revealed differences in promoter preference and requirements for single nucleotide positions. Directed muta- genesis of SASIG1 identified critical residues within the conserved regions 2.4 and 4.2 necessary for binding of the -10 and -35 promoter elements, respectively. SASIG1 and 2, but not SASIG3, each have a typical region 2.5 for binding of the extended -10 promoter element. SASIG3 has a pro-sequence reminiscent of sigma K from Bacillus subtilis, suggesting that proteolytic cleavage from an inactive precursor is involved in the regulation of plastid transcription. In addition, SASIG2 was found to be more abundant in light grown as compared to dark-grown mustard seedlings, while the converse was true for SASIG3. PMID- 12631286 TI - Biotinylation in the hyperthermophile Aquifex aeolicus. AB - Biotin protein ligase (BPL) catalyses the biotinylation of the biotin carboxyl carrier protein (BCCP) subunit of acetyl CoA carboxylase and this post translational modification of a single lysine residue is exceptionally specific. The exact details of the protein-protein interactions involved are unclear as a BPL:BCCP complex has not yet been isolated. Moreover, detailed information is lacking on the composition, biosynthesis and role of fatty acids in hyperthermophilic organisms. We have cloned, overexpressed and purified recombinant BPL and the biotinyl domain of BCCP (BCCP Delta 67) from the extreme hyperthermophile Aquifex aeolicus. In vitro assays have demonstrated that BPL catalyses biotinylation of lysine 117 on BCCP Delta 67 at temperatures of up to 70 degrees C. Limited proteolysis of BPL with trypsin and chymotrypsin revealed a single protease-sensitive site located 44 residues from the N-terminus. This site is adjacent to the predicted substrate-binding site and proteolysis of BPL is significantly reduced in the presence of MgATP and biotin. Chemical crosslinking with 1-ethyl-3-(dimethylamino-propyl)-carbodiimide (EDC) allowed the isolation of a BPL:apo-BCCP Delta 67 complex. Furthermore, this complex was also formed between BPL and a BCCP Delta 67 mutant lacking the lysine residue (BCCP Delta 67 K117L) however, complex formation was considerably reduced using holo-BCCP Delta 67. These observations provide evidence that addition of the biotin prosthetic group reduces the ability of BCCP Delta 67 to heterodimerize with BPL, and emphasizes that a network of interactions between residues on both proteins mediates protein recognition. PMID- 12631288 TI - The kinetic properties of various R258 mutants of deacetoxycephalosporin C synthase. AB - Site-directed mutagenesis was used to investigate the control of 2-oxoacid cosubstrate selectivity by deacetoxycephalosporin C synthase. The wild-type enzyme has a requirement for 2-oxoglutarate and cannot efficiently use hydrophobic 2-oxoacids (e.g. 2-oxohexanoic acid, 2-oxo-4-methyl-pentanoic acid) as the cosubstrate. The following mutant enzymes were produced: R258A, R258L, R258F, R258H and R258K. All of the mutants have broadened cosubstrate selectivity and were able to utilize hydrophobic 2-oxoacids. The efficiency of 2-oxoglutarate utilization by all mutants was decreased as compared to the wild-type enzyme, and in some cases activity was abolished with the natural cosubstrate. PMID- 12631289 TI - Improved ecdysone receptor-based inducible gene regulation system. AB - To develop an ecdysone receptor (EcR)-based inducible gene regulation system, several constructs were prepared by fusing DEF domains of Choristoneura fumiferana EcR (CfEcR), C. fumiferana ultraspiracle (CfUSP), Mus musculus retinoid X receptor (MmRXR) to either GAL4 DNA binding domain (DBD) or VP16 activation domain. These constructs were tested in mammalian cells to evaluate their ability to transactivate luciferase gene placed under the control of GAL4 response elements and synthetic TATAA promoter. A two-hybrid format switch, where GAL4 DBD was fused to CfEcR (DEF) and VP16 AD was fused to MmRXR (EF) was found to be the best combination. It had the lowest background levels of reporter gene activity in the absence of a ligand and the highest level of reporter gene activity in the presence of a ligand. Both induction and turn-off responses were fast. A 16-fold induction was observed within 3 h of ligand addition and increased to 8942-fold by 48 h after the addition of ligand. Withdrawal of the ligand resulted in 50% and 80% reduction in reporter gene activity by 12 h and 24 h, respectively. PMID- 12631290 TI - Distribution of class I, III and IV alcohol dehydrogenase mRNAs in the adult rat, mouse and human brain. AB - The localization of different classes of alcohol dehydrogenases (ADH) in the brain is of great interest because of their role in both ethanol and retinoic acid metabolism. Conflicting data have been reported in the literature. By Northern blot and enzyme activity analyses only class III ADH has been detected in adult brain specimens, while results from riboprobe in situ hybridization indicate class I as well as class IV ADH expression in different regions of the rat brain. Here we have studied the expression patterns of three ADH classes in adult rat, mouse and human tissues using radioactive oligonucleotide in situ hybridization. Specificity of probes was tested on liver and stomach control tissue, as well as tissue from class IV ADH knock-out mice. Only class III ADH mRNA was found to be expressed in brain tissue of all three investigated species. Particularly high expression levels were found in neurons of the red nucleus in human tissue, while cortical neurons, pyramidal and granule cells of the hippocampus and dopamine neurons of substantia nigra showed moderate expression levels. Purkinje cells of cerebellum were positive for class III ADH mRNA in all species investigated, whereas granular layer neurons were positive only in rodents. The choroid plexus was highly positive for class III ADH, while no specific signal for class I or class IV ADH was detected. Our results thus support the notion that the only ADH expressed in adult mouse, rat and human brain is class III ADH. PMID- 12631291 TI - Molecular characterization and allergenic activity of Lyc e 2 (beta fructofuranosidase), a glycosylated allergen of tomato. AB - Until now, only a small amount of information is available about tomato allergens. In the present study, a glycosylated allergen of tomato (Lycopersicon esculentum), Lyc e 2, was purified from tomato extract by a two-step FPLC method. The cDNA of two different isoforms of the protein, Lyc e 2.01 and Lyc e 2.02, was cloned into the bacterial expression vector pET100D. The recombinant proteins were purified by electroelution and refolded. The IgE reactivity of both the recombinant and the natural proteins was investigated with sera of patients with adverse reactions to tomato. IgE-binding to natural Lyc e 2 was completely inhibited by the pineapple stem bromelain glycopeptide MUXF (Man alpha 1-6(Xyl beta 1-2)Man beta 1-4GlcNAc beta 1-4(Fuc alpha 1-3)GlcNAc). Accordingly, the nonglycosylated recombinant protein isoforms did not bind IgE of tomato allergic patients. Hence, we concluded that the IgE reactivity of the natural protein mainly depends on the glycan structure. The amino acid sequences of both isoforms of the allergen contain four possible N-glycosylation sites. By application of MALDI-TOF mass spectrometry the predominant glycan structure of the natural allergen was identified as MMXF (Man alpha 1-6(Man alpha 1-3)(Xyl beta 1-2)Man beta 1-4GlcNAc beta 1-4(Fuc alpha 1-3) GlcNAc). Natural Lyc e 2, but not the recombinant protein was able to trigger histamine release from passively sensitized basophils of patients with IgE to carbohydrate determinants, demonstrating that glycan structures can be important for the biological activity of allergens. PMID- 12631292 TI - Transactivation properties of c-Myb are critically dependent on two SUMO-1 acceptor sites that are conjugated in a PIASy enhanced manner. AB - The transcription factor v-Myb is a potent inducer of myeloid leukemias, and its cellular homologue c-Myb plays a crucial role in the regulation of hematopoiesis. Recently, Bies and coworkers (Bies, J., Markus, J. & Wolff, L. (2002) J. Biol. Chem, 277, 8999-9009) presented evidence that murine c-Myb can be sumoylated under overexpression conditions in COS7 cells when cotransfected with FLAG-tagged SUMO-1. Here we provide independent evidence that human c-Myb is also subject to SUMO-1 conjugation under more physiological conditions as revealed by coimmunoprecipitation analysis of Jurkat cells and transfected CV-1 cells. Analysis in an in vitro conjugation system showed that modification of the two sites K503 and K527 is interdependent. A two-hybrid screening revealed that the SUMO-1 conjugase Ubc9 is one of a few major Myb-interacting proteins. The moderate basal level of sumoylation was greatly enhanced by cotransfection of PIASy, an E3 ligase for SUMO-1. The functional consequence of abolishing sumoylation was enhanced activation both of a transiently transfected reporter gene and of a resident Myb-target gene. When single and double mutants were compared, we found a clear correlation between reduction in sumoylation and increase in transcriptional activation. Enhancing sumoylation by contransfection of PIASy had a negative effect on both Myb-induced and basal level reporter activation. Furthermore, PIASy caused a shift in nuclear distribution of c-Myb towards the insoluble matrix fraction. We propose that the negative influence on transactivation properties by the negative regulatory domain region of c-Myb depends on the sumoylation sites located here. PMID- 12631293 TI - Assessment of porcine and human 16-ene-synthase, a third activity of P450c17, in the formation of an androstenol precursor. Role of recombinant cytochrome b5 and P450 reductase. AB - Recently, we have shown that the biosynthesis of androstenol, a potential endogenous ligand for the orphan receptors constitutive androstane receptor and pregnane-X-receptor, requires the presence of enzymes of the steroidogenic pathway, such as 3 beta-hydroxysteroid dehydrogenase, 5 alpha-reductase and 3 alpha-hydroxysteroid dehydrogenase. In this report, we examine at the molecular level whether the enzyme 17 alpha-hydroxylase/17,20-lyase (P450c17), which possesses dual 17 alpha-hydroxylase and 17,20-lyase activities and catalyzes the production of precursors for glucocorticoids and sex steroids, is also able to catalyze the formation of a third class of active steroids, 16-ene steroids (including androstenol). The role of components of the P450 complex is also assessed. We transfected human embryonic kidney (HEK-293) cells with various amounts of vectors expressing P450c17, NADPH-cytochrome P450 reductase, and cytochrome b5. Our results showed that P450c17 possesses a 16-ene-synthase activity able to transform pregnenolone into 5,16-androstadien-3 beta-ol, without the formation of the precursor 17-hydroxypregnenolone. Cytochrome b5 has a much stronger effect on the 16-ene-synthase activity than on the 17 alpha hydroxylase/17,20-lyase activities. On the other hand, P450reductase has a drastic effect on the latter, but a negligible one on 5,16-androstadien-3 beta-ol synthesis. Our results therefore demonstrate that human P450c17, as other enzymes of the classical steroidogenic pathway, is involved in the biosynthetic pathway leading to the formation of androstenol. PMID- 12631294 TI - Purification of a plant nucleotide pyrophosphatase as a protein that interferes with nitrate reductase and glutamine synthetase assays. AB - An activity that inhibited both glutamine synthetase (GS) and nitrate reductase (NR) was highly purified from cauliflower (Brassica oleracea var. botrytis) extracts. The final preparation contained an acyl-CoA oxidase and a second protein of the plant nucleotide pyrophosphatase family. This preparation hydrolysed NADH, ATP and FAD to generate AMP and was inhibited by fluoride, Cu2+, Zn2+ and Ni2+. The purified fraction had no effect on the activity of NR when reduced methylviologen was used as electron donor instead of NADH; and inhibited the oxidation of NADH by both spinach NR and an Escherichia coli extract in a time-dependent manner. The apparent inhibition of GS and NR and the ability of ATP and AMP to relieve the inhibition of NR can therefore be explained by hydrolysis of nucleotide substrates by the nucleotide pyrophosphatase. We have no evidence that the nucleotide pyrophosphatase is a specific physiological regulator of NR and GS, but suggest that nucleotide pyrophosphatase activity may underlie some confusion in the literature about the effects of nucleotides and protein factors on NR and GS in vitro. PMID- 12631296 TI - Risk factors for pressure ulcers among elderly hip fracture patients. AB - The purpose of this study was to estimate the incidence of hospital-acquired pressure ulcers among elderly patients hospitalized for hip fracture surgery and to identify extrinsic factors that are associated with increased risk. We conducted a secondary analysis of data abstracted from medical records at 20 hospitals in Pennsylvania, Texas, New Jersey, and Virginia. Participants were patients aged 60 years and older admitted with hip fracture to the study hospitals between 1983 and 1993. The incidence of hospital-acquired pressure ulcers was 8.8% (95% confidence interval 8.2%-9.4%). After adjusting for confounding variables, longer wait before surgery, intensive care unit stay, longer surgical procedure, and general anesthesia were significantly associated with higher pressure ulcer risk. Extrinsic factors may be important markers for high pressure ulcer risk in hospitalized hip fracture patients. Although it is not possible to eliminate factors such as requiring an intensive care unit stay or having a long surgical procedure, it may be possible to develop interventions that minimize pressure ulcer risk in patients who experience these factors. PMID- 12631297 TI - Treatment of periprosthetic soft tissue infection of the groin following vascular surgical procedures by means of a polyvinyl alcohol-vacuum sponge system. AB - Deep groin infections after prosthetic vascular surgical procedures represent a serious complication of surgical practice. Septicemia and/or erosive hemorrhage can both be consequences. In this situation, removal of the graft appears to be the only option. However, if the infection is detected early (type Szilagyi III), local treatment to eradicate the infection could serve as an alternative. Twenty four patients with confirmed infection of the soft tissue adjacent to the prosthetic material in the groin were treated locally by implantation of a vacuum sponge system. Duration of this treatment was 2 weeks. All patients showed excellent tissue granulation of the wound area and the microbial stains were negative at the end of therapy. In 21 patients the wound could be primarily closed after explantation of the sponge. Three patients underwent open treatment because of a skin defect. After 12 months, the wounds had healed well in all patients. Histologic evaluation revealed a physiological healing process. Deep soft tissue infections of the groin adjacent to prosthetic vascular material (type Szilagyi III) can be treated effectively and safely with the vacuum sponge system. The treatment is inexpensive, easy to perform, and the initial vascular reconstruction can be preserved. PMID- 12631298 TI - Elevated monocyte chemoattractant protein-1 levels following thermal injury precede monocyte recruitment to the wound site and are controlled, in part, by tumor necrosis factor-alpha. AB - In previous studies, mice given a full-thickness scald injury had an influx of neutrophils into the skin that followed a local increase in a neutrophil chemoattractant. Because macrophages are known to infiltrate the wound area after neutrophils and are essential for normal wound repair, studies were designed to characterize the time course of macrophage accumulation in the wound and to identify the factor(s) responsible for this influx. A macrophage infiltrate into the wound was observed at 4 days post-injury and persisted through at least 10 days. This influx was preceded by an initial fourfold increase in dermal monocyte chemoattractant protein-1 levels at 24 hours post-injury (p < 0.05). This elevation in monocyte chemoattractant protein-1 was enhanced at 4 and 10 days postburn resulting in a sixfold increase over baseline (p < 0.01). Levels of tumor necrosis factor-alpha, a proinflammatory cytokine known to induce chemokine production, were elevated at 90 minutes after injury in burn- versus sham-injured groups (p < 0.05). Furthermore, administration of tumor necrosis factor-alpha neutralizing antibody in vivo reduced the dermal levels of monocyte chemoattractant protein-1 seen at 10 days postburn by 57% (p < 0.01); however, macrophage accumulation was not altered. Thus, elevated systemic TNF-alpha levels may influence the local chemokine milieu following burn injury. PMID- 12631299 TI - Increased expression of interferon-inducible protein-10 during surgically induced peritoneal injury. AB - Interferon-inducible protein 10 (IP-10) is a key regulator of neutrophils, monocytes, and lymphocytes, cells infiltration whose secretory products play a key role in peritoneal wound healing. The objective of the present study was to determine whether IP-10 is expressed by parietal peritoneum and whether its temporal and spatial expression is altered following surgically induced peritoneal injury during healing. Peritoneal sidewall injuries were induced in mice (N = 60), and the severity of adhesions were graded at 12 hours and 1, 2, 4, and 7 days postsurgery. After collection of peritoneal washes, the injured peritoneum with associated adhesion and intact parietal peritoneum were collected to determine IP-10 mRNA and protein expression using quantitative reverse transcription-polymerase chain reaction, enzyme-linked immunosorbent assay, and immunohistochemistry. Peritoneal injury resulted in adhesion formation with increased severity by day 7 postsurgery. The intact parietal peritoneum expressed IP-10 mRNA, whose level of expression significantly increased following peritoneal injury and reached a maximum at day 4 (p = 0.001), declining to the uninjured control levels by day 7 post-injury. The level of IP-10 in peritoneal washes also increased as a result of peritoneal injury. Immunohistochemically, IP 10 was localized to various inflammatory and immune cells, adhesion fibroblasts, and mesothelial cells, and its intensity increased during the course of wound healing. In conclusion, we showed that parietal peritoneum expresses IP-10 and peritoneal tissue injury results in an elevated level of its expression throughout the early phase of wound healing. The results suggest that IP-10 and its elevated expression may play a role in peritoneal cellular activities that influence the early phases of tissue repair and, possibly, the development of peritoneal adhesions. PMID- 12631300 TI - Effects of vascular endothelial growth factor on wound closure rates in the genetically diabetic mouse model. AB - Impaired wound healing is characteristic of diabetic patients. Potential reasons include poor inflammatory response, granulation tissue formation, and abnormal patterns of cytokine release and response. Vascular endothelial growth factor, abnormally regulated during healing in diabetics, is the major factor stimulating angiogenesis during normal wound healing. We tested our hypothesis that topically applied vascular endothelial growth factor would improve wound closure rates in diabetic animals in a full-thickness wound model in genetically diabetic mice (C57 BL/KsJ db/db). Animals received either 1.0 micro g of vascular endothelial growth factor165 or polyethylene glycol alone topically to wounds daily between days 0 and 4 post-wounding. Wound area was measured at days 0, 5, 10, 15, and 21. Data were analyzed using probit analysis and expressed as length-of-time (LT) to 50, 90, and 95% wound closure. Among untreated animals, nondiabetics had an LT50 of 8.5 days (fiducial limits 8.3-8.7), while diabetics had an LT50 of 15.8 days (15.6-16.1). Vascular endothelial growth factor-treated animals had LT50 values of 7.8 (7.6-8.1) and 11.8 days (11.6-12.0) for nondiabetics and diabetics, respectively, representing a 25% improvement in time to 50% closure in treated diabetics. We conclude that topically applied vascular endothelial growth factor improves time to wound closure in the genetically diabetic mouse model. PMID- 12631301 TI - Upward migration of cultured autologous keratinocytes in Integra artificial skin: a preliminary report. AB - The combination of cultured autologous keratinocytes with the dermal regeneration template Integra could offer increased possibilities for reconstructive surgery and wound healing. A single-step application of cells, centrifuged deep into an Integra-like matrix at the silicone-matrix junction, has been described but might prove technically complex for clinical use. We have investigated the possibility of simplifying this procedure by applying cultured cells directly to the underside of the Integra or directly to the wound bed immediately prior to grafting. The objective was to see whether cells would migrate through the matrix in an upward direction. We tested the principle of this concept using a pig wound healing model. Integra was seeded directly with cultured cells and grafted onto fresh full-thickness wounds, or unseeded Integra was applied to freshly excised wound beds that had just been seeded with the same number of cells. Biopsies were taken at 3, 7, 11, and 14 days. Histological sections showed that the cells moved through the Integra to give a confluent surface epithelium. Direct seeding onto the Integra was the most efficient method. Transduction of cultured autologous keratinocytes in vitro with a MFGlacZnls retrovirus confirmed that the epidermis was derived from the cultured autologous keratinocytes. PMID- 12631302 TI - Hyperbaric oxygen treatment prevents up-regulation of angiogenesis following partial-thickness skin grafts in the pig. AB - We hypothesized that hyperbaric oxygen treatment (HBO) would reduce neovascularization following partial-thickness skin grafting in the Yucatan pig. Results show that capillary density 4 days post-graft is increased twofold in grafts not treated with HBO, compared to normal, ungrafted skin or skin grafts from pigs treated with HBO, p < 0.05. Similarly, the expression of vascular endothelial growth factor, a growth factor associated with neovascularization, was also reduced by HBO. Cell density in the graft boundary increased gradually after grafting, reaching a maximum 2.7-fold increase, relative to normal skin, at day 4, p < 0.05. Cell nuclei positive for proliferating cell nuclear antigen, a marker of proliferation, increased threefold by day 2, p < 0.05, and then declined to normal levels by day 7. HBO had no effect on cell density or proliferation in the boundary region or on shear strength of the graft. In the epidermis, proliferation declined 80% 2 days after grafting and then returned to levels observed in normal skin by day 4, p < 0.05; however, in pigs treated with HBO, we observed no decline in proliferation. These findings confirm the hypothesis that HBO reduces neovascularization in the partial-thickness skin graft while preserving regenerative capacity in the graft boundary and normal proliferative capacity of the epidermis. PMID- 12631303 TI - Sterile isolation and cryopreservation of human adherent monocytes/macrophages from mononuclear cell apheresis preparations. AB - There is evidence from animal and human study that suggests clinical use of monocytes/macrophages may be of benefit in wound healing, tissue regeneration, and cancer therapy. To facilitate further study, a method was developed for sterile isolation and cryopreservation of adherent monocyte/macrophages from mononuclear cell apheresis units collected from unstimulated normal human donors. Preparations contained approximately 1 x 108 total cells and were comprised of approximately 60% monocytes, 38% lymphocytes, and 2% granulocytes. Cells could be cryopreserved for up to 8 months and subsequently thawed and stored at 1-6 degrees C for up to 4 hours with retention of viability and adequate phagocytic function. These cells can be used in clinical trials to determine their possible therapeutic benefit, e.g., whether administration of exogenously supplied cells improves the healing of chronic wounds or promotes the regeneration of transected nerves. PMID- 12631306 TI - Water supply and sanitation: some misconceptions. PMID- 12631304 TI - Standardized qualitative evaluation of scar tissue properties in an animal wound healing model. AB - There is a great need to establish reproducible methods for evaluative studies of wound treatment and wound healing. Validation of the healing process through optical techniques, as well as histologic and immunohistochemical methodologies, have been improved and to some extent have become well-established assays. Data relating to biomechanical properties, e.g., evaluation of the tensile strength of scar tissue that forms in experimental wound treatment strategies, are less widely available. We chose the domestic pig as an animal model in which to examine epidermal wound healing. We implanted specially made chambers that served to isolate the wounds and prevent epidermal migration from the edges. We performed histologic and immunohistochemical analyses as well as evaluation of biomechanical qualities of scar tissue using laser tensiometry. Pig skin is well suited for wound healing studies, and wound creation, implantation of the chambers, and the regular changing of dressings could all be carried out in the operating theater. In addition to established macroscopic evaluation and microscopic documentation, the need for objective biomechanical assessment of scar tissue by measuring tensile strength has been met using laser tensiometry. By optimizing methods for measuring tensile strength, it is possible to evaluate the biomechanical quality of scar tissue formed following different courses of wound treatment, as well as histologic assessment. PMID- 12631307 TI - A health facility based case-control study of effectiveness of insecticide treated nets: potential for selection bias due to pre-treatment with chloroquine. AB - Case control studies offer an attractive way to assess the effectiveness of insecticide treated nets (ITN) under programme conditions but have the drawback of being susceptible to bias in the choice of controls. We evaluated the potential for pre-treatment with chloroquine to result in misclassification of cases and controls and affect estimates of ITN effectiveness in case control studies in urban and rural clinics in Eastern Afghanistan. During the one-month study, use of ITN showed no effect against malaria in the urban clinic (adjusted odds ratio OR 1.08; 95% CI 0.73-1.6) and the protective effect seen in the rural clinic was not significant (OR 0.62; 95% CI 0.2-2.4). Levels of pre-treatment were high in both clinics: 24% in urban and 19% in rural clinic attenders. In the urban clinic attenders the level of pre-treatment between bed net users and non users was not significantly different (OR 1.07, 95% CI 0.70-1.64); therefore the misclassification of cases as controls did not introduce any selection bias. Amongst rural clinic attenders, bed net users were less likely to pre-treat with chloroquine than users (OR 0.33, 95% CI 0.14-0.77); this introduced a selection bias that resulted in an underestimation of the effectiveness of bed nets. Case control studies using health facility data are liable to selection bias especially in areas of high pre-treatment rates with chloroquine. Generalisation of results over a wide geographic region, or between urban and rural settings, may not be appropriate. PMID- 12631308 TI - Clinical efficacy of chloroquine in young children with uncomplicated falciparum malaria--a community-based study in rural Burkina Faso. AB - We report on a 14-day study on the efficacy of chloroquine for treating uncomplicated falciparum malaria in young children of a malaria holoendemic area in rural Burkina Faso. In this community-based study, the overall treatment failure rate was 12/120 (10%), with no differences between villages. This supports the evidence for a still sufficient efficacy of chloroquine in north western Burkina Faso. PMID- 12631309 TI - Evaluation of efficacy of community-based vs. institutional-based direct observed short-course treatment for the control of tuberculosis in Kilombero district, Tanzania. AB - Tuberculosis (TB) has reappeared as a serious public health problem. Non compliance to antituber-culous drug treatment is cited as one of the major obstacles to the containment of the epidemic. Compliance may be optimized by Directly Observed Treatment (DOT) and short-course treatment regimens. Since 1986, Tanzanian TB patients have received daily DOT at health facilities for the first 2 months of the treatment course. However, adherence and cure rates have been falling as the number of TB cases continues to increase and the burden on already stretched health facilities threatens to become unmanageable. We used an open cluster randomized controlled trial to compare community-based DOT (CBDOT) using a short-course drug regimen with institutional-based DOT (IBDOT). A total of 522 (301 IBDOT and 221 CBDOT) patients with sputum-positive TB were recruited. Overall, there was no significant difference in conversion and cure rates between the two strategies [M-H pooled odds ratio (OR) 0.62; 95% confidence interval (CI) 0.23, 1.71 and OR = 1.58; 95% CI 0.32, 7.88, respectively] suggesting that CBDOT may be a viable alternative to IBDOT. CBDOT may be particularly useful in parts of the country where people live far from health facilities. PMID- 12631310 TI - Tuberculosis treatment in complex emergencies: are risks outweighing benefits? AB - Tuberculosis (TB) is a major public health problem in complex emergencies. Humanitarian agencies usually postpone the decision to offer TB treatment and opportunities to treat TB patients are often missed. This paper looks at the problem of tuberculosis treatment in these emergencies and questions whether treatment guidelines could be more flexible than international recommendations. A mathematical model is used to calculate the risks and benefits of different treatment scenarios with increasing default rates. Model outcomes are compared to a situation without treatment. An economic analysis further discusses the findings in a trade-off between the extra costs of treating relapses and failures and the savings in future treatment costs. In complex emergencies, if a TB programme could offer 4-month treatment for 75% of its patients, it could still be considered beneficial in terms of public health. In addition, the proportion of patients following at least 4 months of treatment can be used as an indicator to help evaluate the public health harm and benefit of the TB programme. PMID- 12631311 TI - Clinical presentation of subacute sclerosing panencephalitis in Papua New Guinea. AB - Eighty-three children presented at Goroka Base Hospital in the Eastern Highlands Province (EHP) of Papua New Guinea over a period of 3 years and 9 months between February 1997 and November 2000 were confirmed to have subacute sclerosing panencephalitis (SSPE). Confirmation of the diagnosis was based on the demonstration of high titres of measles antibodies in the cerebrospinal fluid and/or serum in association with clinical features supportive of SSPE, including characteristic electroencephalographic changes and amplification of measles virus genome by reverse transcriptase polymerase chain reaction in some cases. The mean cerebrospinal fluid and serum enzyme immunoassay antibody levels among the SSPE patients were 38 250 and 860 580, respectively. The mean age of onset of SSPE was 7.9 +/- 2.6 years and ranged between 2 and 14 years. The overall male to female ratio was 1.2:1 and 1.4:1 for EHP. PMID- 12631312 TI - Clinical findings among 62 Thais with cholangiocarcinoma. AB - Cholangiocarcinoma, a malignancy of the biliary duct system, tends to grow slowly and to infiltrate the walls of the ducts, dissecting along tissue planes and leading to biliary tract obstruction. In a retrospective case review, 62 cases diagnosed with cholangiocarcinoma between January 1997 and December 2001 at the King Chulalongkorn Memorial Hospital in Bangkok were studied. The most commonly observed clinical symptoms and signs were fever (96.8%), abdominal pain (87.1%) and malaise or weakness (83.9%). Fifty-one of the 62 tumours detected were distal extrahepatic (82.3%), seven perihilar (11.3%) and four intrahepatic (6.7%). There was a significant increase in serum bilirubin and marked elevation of serum alkaline phosphatase. Almost half (45.2%) of the patients presented with other pathological conditions, mostly cholangitis (19 cases) resulting from acute biliary tract obstruction and ascites (13 cases). The pathogens detected were similar in cases of cholangitis alone and cholangiocarcinoma. The major risk factor for this cancer in Thailand is believed to be exposure to liver fluke in insufficiently cooked traditional foods. As it is still endemic in Thailand, continuous prevention and surveillance of this public health problem are necessary. PMID- 12631313 TI - Hepatic ultrasound in a population with high incidence of invasive amoebiasis: evidence for subclinical, self-limited amoebic liver abscesses. AB - About 10% of successfully treated amoebic liver abscesses (ALA) do not completely resolve and can be detected by ultrasound as typical residual liver lesions. The frequency of these residues should be an indicator for the prevalence of ALA in a given population, and may help to solve the question whether non-clinical, self healing ALAs occur. We have performed hepatic ultrasound in 1036 adult individuals living in a high-risk area for ALA in Central Vietnam and identified typical ALA residual lesions in about 1.2% of the subjects. As expected, these lesions were associated with positive amoeba serology and were found in 11.9% of individuals with a previous ALA history. However, more than 50% of the residues were identified in individuals who had never developed symptoms suspected for ALA and who never received any ALA specific treatment, suggesting that subclinical, self-limited hepatic amoeba abscesses truly exist. PMID- 12631314 TI - Overview of injection practices in two governorates in Egypt. AB - OBJECTIVE: To describe the extent and characteristics of injection use and injection providers in Egypt, given that unsafe injections are associated with blood-borne pathogen transmission. METHODS: Household surveys of a population based sample of residents in the Nile Delta and in Upper Egypt; focus group discussions and in-depth interviews with community target groups, formal and informal medical providers. RESULTS: Of 4197 persons interviewed, 26.2% reported receiving an injection in the past 3 months. Of these, 77% reported it was for therapeutic indications. The age-sex specific prevalence of injections was highest among children 0-2 years of age and among older adults. Women were more likely to report having an injection than men, particularly at the age above 20 years. Overall, respondents reported receiving on average 4.2 injections per year, indicating that up to 281 million injections are provided per year in Egypt. Injection administrators were public and private sector physicians, pharmacists, barbers, doctor assistants, housekeepers, relatives and friends. Injection prescribers were mostly private and public sector physicians. Of the 1101 respondents who received an injection in the past 3 months, 92 (8.4%) reported that the provider did not use a syringe taken from a closed sealed packet. CONCLUSION: The frequency of therapeutic injection use is high in Egypt and may contribute to blood-borne pathogen transmission. The Ministry of Health and Population (MOHP) is developing interventions targeted towards promotion of injection safety and reduction of injection overuse on community basis as part of a comprehensive strategy to prevent blood-borne pathogen transmission in Egypt. PMID- 12631315 TI - Hospital costs of high-burden diseases: malaria and pulmonary tuberculosis in a high HIV prevalence context in Zimbabwe. AB - This paper explores the measurement of hospital costs and efficiency in a context where data is scarce, incomplete or of poor quality. It argues that there is scope for using tracers to examine and compare hospital cost structures and relative efficiency in such contexts. Two high-burden diseases, malaria and pulmonary tuberculosis, are used as tracers to calculate the average costs of inpatient care at selected tertiary hospitals. This study shows that it is feasible to prospectively collect cost data for specific diseases and explore in detail both patient cost distribution and susceptible areas for efficiency improvement. The present study found that the critical source of efficiency variation in public hospitals in Zimbabwe lies in the way hospital beds are used. PMID- 12631317 TI - [Estacionalidad de parasitos intestinales en suelos periurbanos de la ciudad de Neuquen, Patagonia, Argentina]. AB - OBJECTIVE: To determine the presence of eggs, larva, cysts and oocysts of intestinal parasites in the soil of a suburb of Neuquen city during 1 year in order to evaluate their seasonal fluctuations in relation to climatic data and soil characteristics in the studied area. METHODS: A total of 107 soil samples were processed for parasite isolation by sedimentation and flotation methods during the four seasons of the year. Meteorological data were registered and physical, chemical and structural characteristics of the soil were analysed. RESULTS: About 28.9% of the soil samples were positive for at least one parasite form. Six protozoa species (cysts of Entamoeba sp., Enteromonas sp., Endolimax sp., Giardia sp., Iodamoeba sp. and coccidia oocysts) were recovered, but neither larvae nor eggs of human or animal helminths parasites were detected. The percentage of contaminated soil samples and the diversity of species showed a marked decrease in the warm and dry months of the summer. The soil was sandy, without vegetable cover, well drained, and with scarce organic matter content. CONCLUSION: The frequencies of parasite recovery and the number of species show seasonal fluctuations related to the rainfall. The importance of soil as a risk factor for the transmission of intestinal parasites in the studied area is conditioned by its structural characteristics, which prevent retaining the humidity, and by climatic variables. The interrelation of both factors determines unfavourable conditions that could explain the low level of contamination observed in soil as well as the absence of eggs and helminth larvae. PMID- 12631316 TI - Evidence-based treatment guidelines for sexually transmitted infections developed with and for female sex workers. AB - BACKGROUND: Sex work is frequently one of the few options women in low-income countries have to generate income for themselves and their families. Treating and preventing sexually transmitted infections (STIs) among sex workers (SWs) is critical to protect the health of the women and their communities; it is also a cost-effective way to slow the spread of HIV. Outside occasional research settings however, SWs in low-income countries rarely have access to effective STI diagnosis. OBJECTIVES: To develop adequate, affordable, and acceptable STI control strategies for SWs. METHODS: In collaboration with SWs we evaluated STIs and associated demographic, behavioural, and clinical characteristics in SWs living in two cities in Madagascar. Two months post-treatment and counselling, incident STIs and associated factors were determined. Evidence-based STI management guidelines were developed with SW representatives. RESULTS: At baseline, two of 986 SWs were HIV+; 77.5% of the SWs in Antananarivo and 73.5% in Tamatave had at least one curable STI. Two months post-treatment, 64.9% of 458 SWs in Antananarivo and 57.4% of 481 women in Tamatave had at least one STI. The selected guidelines include speculum exams; syphilis treatment based on serologic screening; presumptive treatment for gonorrhoea, chlamydia, and trichomoniasis during initial visits, and individual risk-based treatment during 3-monthly follow-up visits. SWs were enthusiastic, productive partners. CONCLUSIONS: A major HIV epidemic can still be averted in Madagascar but effective STI control is needed nationwide. SWs and health professionals valued the participatory research and decision-making process. Similar approaches should be pursued in other resource-poor settings where sex work and STIs are common and appropriate STI diagnostics lacking. PMID- 12631318 TI - Domiciliation process of Rhodnius stali (Hemiptera: Reduviidae) in Alto Beni, La Paz, Bolivia. AB - We report a systematic collection of Triatominae inside houses and in the peridomestic environment of Alto Beni, department of La Paz, Bolivia. This area is free of Triatoma infestans and although we detected previously seropositivity for Trypanosoma cruzi, the Alto Beni region is not officially considered as endemic for Chagas disease. From 11 houses of five localities, we collected adults, nymphs and eggs of a Rhodnius species, which was confirmed by morphological and morphometric analysis as Rhodnius stali. This little-known species has long been confused with R. pictipes, and was originally described from museum specimens labelled as R. pictipes. Our data show that R. stali is able to establish colonies in domestic and peridomestic habitats in Bolivia, and it is probably the vector responsible for Chagas disease seropositivity observed in the indigenous population of Alto Beni. PMID- 12631319 TI - Ecological and molecular studies on emerging schistosomiasis mansoni in Dhofar Governorate, Sultanate of Oman. AB - The recent detection of some cases of autochtonous schistosomiasis mansoni in Dhofar, Oman, prompted a search for the transmission sites. The five field surveys we conducted from November 2000 to February 2002 provided ecological data on schistosomiasis in Dhofar. Twenty-eight water bodies situated within 8-160 km from Salalah, the largest city of Dhofar and at altitudes of up to 900 m, were surveyed for freshwater snails. Biomphalaria arabica was found in 15 of them. Three sites (Tibraq, Siginitti and Arazat) had Schistosoma infected snails, the first snails shedding cercariae of this parasite ever collected in Oman. The parasite from Dhofar was analysed by Random Amplified Polymorphic DNA comparisons using 11 primers and 167 polymorphic fragments and had 87-88% similarity with Schistosoma mansoni from Guadeloupe, but only 37-38% similarity with S. rodhaini from Burundi. Thus, it is a strain of S. mansoni. During the November 2000 survey, the prosobranch snail Melanoides tuberculata was associated with B. arabica in 10 of the 13 B. arabica sites. Cercariae from other species of Digenea emerged from five of the B. arabica sites, including the three named above. This paper presents the first finding of S. mansoni in the Dhofar Governorate and represents an initial study of the biology of S. mansoni transmission. This parasite and its cycle need further biological and molecular characterization, and the clarification of its epidemiological status in Dhofar Governorate is an urgent task. PMID- 12631320 TI - Prospective evaluation and comparison of the direct agglutination test and an rK39-antigen-based dipstick test for the diagnosis of suspected kala-azar in Nepal. AB - The diagnosis of visceral leishmaniasis (kala-azar) remains difficult in rural endemic areas and practical and reliable tests are badly needed. Two serological tests, the Direct Agglutination Test (DAT) and an rK39-antigen-based dipstick test, were compared to parasitological diagnosis in a group of 184 patients presenting at a tertiary care centre in south-eastern Nepal with a history of fever > or = 14 days and splenomegaly; 139 patients had a parasitologically proven kala-azar and 45 patients had a negative parasitological work-up. The rK39 dipstick showed a sensitivity of 97% and a specificity of 71%. The DAT was up to 99% sensitive with a low cut-off titre (1:400) but its specificity did not exceed 82% even with a high cut-off titre (1:51 200). Both tests could be used for screening suspect patients in endemic areas. However, their use as confirmatory tests should be restricted to situations where the proportion of kala-azar among clinical suspect patients is high. The rK39 dipstick is cheaper and easier to use than the DAT and could be used widely provided that both its performance and production remain stable. PMID- 12631321 TI - Internal telomeric repeats and 'TCP domain' protein-binding sites co-operate to regulate gene expression in Arabidopsis thaliana cycling cells. AB - We have focused our interest on two cis-regulatory elements, named site II motif and telo box, identified within the promoter of plant proliferating cellular nuclear antigen (PCNA) and putatively involved in meristematic expression of the gene. A conserved topological association between site II motifs and telo boxes is observed in the promoter of numerous genes expressed in cycling cells, including several cell cycle-related genes and 153 Arabidopsis genes encoding ribosomal proteins. Meristematic expression of a GUS reporter gene was observed in plants under the control of Arabidopsis site II motif within a minimal promoter. This expression is strongly enhanced by addition of a telo box within this chimaeric promoter. We showed by gel retardation experiments that the site II motif is a target for several DNA-binding activities present in Arabidopsis crude cell extract and can bind a transcription factor, At-TCP20, from the Teosinte branched 1, Cycloidea, PCF (TCP)-domain protein family. In yeast two hybrid experiments, At-TCP20 appears to be a potential partner of AtPuralpha, which was previously shown to bind telo boxes. An important consequence of this analysis is to reveal new and conserved regulatory processes concerning the regulation of plant ribosomal gene expression in cycling cells. The implication of these observations in plant-specific developmental pathways is discussed. PMID- 12631322 TI - Methylation of histone H3 in euchromatin of plant chromosomes depends on basic nuclear DNA content. AB - Strong methylation of lysine 4 (K4) and low methylation of lysine 9 (K9) have been proposed as modifications of histone H3, typical for transcriptionally active euchromatin and the opposite for inactive heterochromatin. We have analysed the correlation between the global distribution of histone H3, methylated at either lysine 4 or lysine 9, and of microscopically detectable euchromatic or heterochromatic regions in relation to genome size for 24 plant species. Two different distribution patterns of methylated (K9)H3 (Met(K9)H3) were found that depend on genome size. For most species with small genomes (1C <500 Mbp), including Arabidopsis thaliana, strong methylation of (K9)H3 was restricted to constitutive heterochromatin. Species with larger genomes showed a uniform distribution of Met(K9)H3. Contrary to this and regardless of the genome size, methylated (K4)H3 (Met(K4)H3) was found to be enriched within the euchromatin of all species. Transcriptionally less active B chromosomes showed the same patterns as basic A chromosomes. We thus propose that large genomes with high amounts of dispersed repetitive sequences (mainly retroelements) have to silence these sequences and therefore display epigenetic modifications such as methylation of DNA and (K9)H3 also within euchromatic regions. PMID- 12631324 TI - Growth and stomata development of Arabidopsis hypocotyls are controlled by gibberellins and modulated by ethylene and auxins. AB - The plant hormones gibberellin (GA), ethylene and auxin can promote hypocotyl elongation of Arabidopsis seedlings grown in the light on a low nutrient medium (LNM). In this study, we used hypocotyl elongation as a system to investigate interactions between GA and ethylene or auxin and analysed their influence on the development of stomata in the hypocotyl. When applied together, GA and ethylene or auxin exerted a synergistic effect on hypocotyl elongation. Stimulated cell elongation is the main cause of hypocotyl elongation. Furthermore, hypocotyls treated with GA plus either ethylene or auxin show an increased endoreduplication. In addition, a small but significant increase in cell number was observed in the cortical cell files of hypocotyls treated with ethylene and GA together. However, studies with transgenic seedlings expressing CycB1::uidA genes revealed that cell division in the hypocotyl occurs only in the epidermis and mainly to form stomata, a process strictly regulated by hormones. Stomata formation in the hypocotyl is induced by the treatment with either GA or ethylene. The effect of GA could be strongly enhanced by the simultaneous addition of ethylene or auxin to the growth medium. Gibberellin is the main signal inducing stomata formation in the hypocotyl. In addition, this signal regulates hypocotyl elongation and is modulated by ethylene and auxin. The implication of these three hormones in relation to cell division and stomata formation is discussed. PMID- 12631323 TI - Identification of photorespiratory glutamate:glyoxylate aminotransferase (GGAT) gene in Arabidopsis. AB - In the photorespiratory process, peroxisomal glutamate:glyoxylate aminotransferase (GGAT) catalyzes the reaction of glutamate and glyoxylate to 2 oxoglutarate and glycine. Although GGAT has been assumed to play important roles for the transamination in photorespiratory carbon cycles, the gene encoding GGAT has not been identified. Here, we report that an alanine:2-oxoglutarate aminotransferase (AOAT)-like protein functions as GGAT in peroxisomes. Arabidopsis has four genes encoding AOAT-like proteins and two of them (namely AOAT1 and AOAT2) contain peroxisomal targeting signal 1 (PTS1). The expression analysis of mRNA encoding AOATs and EST information suggested that AOAT1 was the major protein in green leaves. When AOAT1 fused to green fluorescent protein (GFP) was expressed in BY-2 cells, it was found to be localized to peroxisomes depending on PTS1. By screening of Arabidopsis T-DNA insertion lines, an AOAT1 knockout line (aoat1-1) was isolated. The activity of GGAT and alanine:glyoxylate aminotransferase (AGAT) in the above-ground tissues of aoat1-1 was reduced drastically and, AOAT and glutamate:pyruvate aminotransferase (GPAT) activity also decreased. Peroxisomal GGAT was detected in the wild type but not in aoat1 1. The growth rate was repressed in aoat1-1 grown under high irradiation or without sugar, though differences were slight in aoat1-1 grown under low irradiation, high-CO2 (0.3%) or high-sugar (3% sucrose) conditions. These phenotypes resembled those of photorespiration-deficient mutants. Glutamate levels increased and serine levels decreased in aoat1-1 grown in normal air conditions. Based on these results, it was concluded that AOAT1 is targeted to peroxisomes, functions as a photorespiratory GGAT, plays a markedly important role for plant growth and the metabolism of amino acids. PMID- 12631325 TI - Correlated clustering and virtual display of gene expression patterns in the wheat life cycle by large-scale statistical analyses of expressed sequence tags. AB - Compared to rice, wheat exhibits characteristic growth habits and contains complex genome constituents. To assess global changes in gene expression patterns in the wheat life cycle, we conducted large-scale analysis of expressed sequence tags (ESTs) in common wheat. Ten wheat tissues were used to construct cDNA libraries: crown and root from 14-day-old seedlings; spikelet from early and late flowering stages; spike at the booting stage, heading date and flowering date; pistil at the heading date; and seeds at 10 and 30 days post-anthesis. Several thousand colonies were randomly selected from each of these 10 cDNA libraries and sequenced from both 5' and 3' ends. Consequently, a total of 116 232 sequences were accumulated and classified into 25 971 contigs based on sequence homology. By computing abundantly expressed ESTs, correlated expression patterns of genes across the tissues were identified. Furthermore, relationships of gene expression profiles among the 10 wheat tissues were inferred from global gene expression patterns. Genes with similar functions were grouped with one another by clustering gene expression profiles. This technique might enable estimation of the functions of anonymous genes. Multidimensional analysis of EST data that is analogous to the microarray experiments may offer new approaches to functional genomics of plants. PMID- 12631326 TI - Initiation of silencing of maize MuDR/Mu transposable elements. AB - Homology-dependent gene silencing contributes to genomic stability through suppression of transposable elements. Co-ordinate epigenetic silencing is the main regulatory mechanism controlling dispersed, multicopy MuDR/Mu elements responsible for Mutator activity in maize. Silencing eliminates transposition and proceeds through transcriptional inactivation of MuDR genes and DNA methylation of the terminal inverted repeats (TIRs) in both the regulatory MuDR and non autonomous Mu elements. In plants with active MuDR/Mu elements, initiation of silencing coincides with nuclear retention of non-polyadenylated RNA derived from MuDR and recently described MuDR homologs (hMuDR elements). Nuclear accumulation of MuDR/hMuDR RNA is developmentally progressive, paralleling loss of Mutator activity and is predictive of loss of Mu somatic excision in the progeny. A high ratio of nuclear to cytoplasmic RNA is the earliest molecular marker for MuDR silencing suggesting that the nuclear RNA may trigger transcriptional silencing. We also demonstrate the constitutive presence of small transposon-specific RNAs of 21-26 nucleotides in all maize lines tested, independent of the Mutator activity. The role of the small RNAs in transposon silencing and translational regulation of transposon-encoded proteins is discussed. PMID- 12631327 TI - Identification of a mitochondrial transporter for basic amino acids in Arabidopsis thaliana by functional reconstitution into liposomes and complementation in yeast. AB - We describe the identification and functional characterization of two Arabidopsis mitochondrial basic amino acid carriers (BAC), AtmBAC1 and AtmBAC2, which are related to the yeast ornithine (Orn) carrier Ort1p, also known as Arg11p. The arg11 mutant requires arginine (Arg) supplementation because it fails to export sufficient ornithine from the mitochondrion to the cytosol where it is converted to arginine. AtmBAC1 and, to a lesser extent, AtmBAC2 partially replaced the function of Ort1p in yeast arg11. The more efficient putative carrier, AtmBAC1, was expressed in E. coli, purified, and reconstituted into phospholipid vesicles, where it transported the basic l-amino acids arginine, lysine, ornithine and histidine (in order of decreasing affinity). AtmBAC1 recognized l-histidine whereas both yeast Ort1p and the mammalian ortholog ORNT1p do not. Also different from ORNT1p, AtmBAC1 did not transport citrulline. AtmBAC1 appeared to be more stereospecific than the yeast and mammalian ornithine carriers, exhibiting greater preference for the l-forms of arginine, lysine and ornithine. By RT-PCR, both AtmBAC1 and AtmBAC2 transcripts were detected in stems, leaves, flowers, siliques, and seedlings. Expression of AtmBAC1 in seedlings is consistent with its involvement in Arg breakdown in early seedling development, i.e. delivery of Arg to mitochondrial arginase. The Km (0.19 mm) for Arg uptake by AtmBAC1 was close to the value we previously determined for the saturable component of Arg uptake into intact mitochondria from soybean seedling cotyledons. PMID- 12631328 TI - Antisense inhibition of NADH glutamate synthase impairs carbon/nitrogen assimilation in nodules of alfalfa (Medicago sativa L.). AB - Legumes acquire significant amounts of nitrogen for growth from symbiotic nitrogen fixation. The glutamine synthetase (GS)/NADH-dependent glutamate synthase (NADH-GOGAT) cycle catalyzes initial nitrogen assimilation. This report describes the impact of specifically reducing nodule NADH-GOGAT activity on symbiotic performance of alfalfa (Medicago sativa L.). Four independent transgenic alfalfa lines, designated GA89, GA87, GA88, and GA82 (for GOGATantisense), containing an antisense NADH-GOGAT cDNA fragment under the control of the soybean leghemoglobin (lbc3) promoter were evaluated. The GA plants were fertile and showed normal growth in non-symbiotic conditions. The NADH-GOGAT antisense transgene was heritable and the T1 plants showed phenotypic alterations - similar to primary transformants. Clonally propagated plants were inoculated with Sinorhizobium meliloti after rooting and the symbiotic phenotype was analyzed 21 days post-inoculation. Nodules of each GA line had reduced NADH GOGAT activity, ranging from 33 to 87% of control plants, that was accompanied by comparable decreases in RNA and protein. Plants from the GA89 line, with the lowest NADH-GOGAT activity (c. 30%), presented a strikingly altered symbiotic phenotype: concomitantly activities of key enzyme for carbon and nitrogen assimilation decreased; nodule amino acids and amides were reduced while sucrose accumulated. Antisense GOGAT plants were chlorotic, reduced in fresh weight, and had a lower N content than control plants. Photosynthesis was also impaired in antisense plants. Specifically, reducing NADH-GOGAT in nodules resulted in plants having impaired nitrogen assimilation and altered carbon/nitrogen metabolic flux. PMID- 12631329 TI - Double fertilization in maize: the two male gametes from a pollen grain have the ability to fuse with egg cells. AB - In flowering plants, two male gametes from a single pollen grain fuse with two female gametes, the egg and central cells, to form the embryo and endosperm, respectively. The question then arises whether the two male gametes fuse randomly with the egg and central cells. We investigated this question using two nearly isogenic maize lines with supernumerary B chromosomes (TB10L18) or without (r tester). B chromosomes regularly undergo non-disjunction at the second pollen mitosis, producing one sperm cell with zero B chromosomes and one with two. We first confirmed earlier studies showing an excess of transmission of the B chromosomes to the embryo rather than to the endosperm. We then tested the possibility of a directed fertilization. For TB10L18 pollen, we could demonstrate the existence of a size dimorphism between the two sperm cells, correlated to the content in B chromosomes, as detected by fluorescence in situ hybridization (FISH). However, no directed fusion of B chromosome containing sperm to egg cells could be detected when using in vitro fertilization. The absence of directed fusion in vitro could also be demonstrated for control lines. We conclude that both male gametes have the capacity to fuse with the egg cell in maize, although sexual reproduction results in a preferential transmission of supernumerary B chromosomes. PMID- 12631330 TI - In vitro analysis of transcription initiation and termination from the Lhcb1 gene family in Nicotiana sylvestris: detection of transcription termination sites. AB - Genes encoding chlorophyll a/b-binding proteins of photosystem II (Lhcb) constitute a multigene family. Nine Lhcb1 genes have previously been isolated from the tobacco species, Nicotiana sylvestris, and the transcription initiation sites in vivo have been mapped. Reaction conditions from a previously developed in vitro transcription system from tobacco cultured cells were optimized for the Lhcb1 genes. Transcription initiation sites in vitro predominantly coincided with those found in vivo and were typically cytidines, a system unique to N. sylvestris Lhcb1 genes. CTC*A (C* for initiation site) was a consensus motif for the initiation region in vitro, as reported in vivo. Mutation analysis defined functionally that the TATA box is essential for transcription initiation and that the CTCA motif is a determinant of transcription initiation sites but not transcript levels. Polyadenylation sites were determined from in vivo transcripts, located 12-21 nt downstream from likely poly(A) signals. Four major 3'-ends of in vitro transcripts from Lhcb1*6 were detected, 40-300 nt downstream of the poly(A) site, suggestive of multiple, discrete transcription termination sites. These 3'-ends are mapped in or nearby T-rich sequences. PMID- 12631331 TI - A novel protein from Brassica napus has a putative KID domain and responds to low temperature. AB - To identify factors that interact with histone deacetylase (HDAC) in Brassica napus, a yeast two-hybrid library was screened using the Arabidopsis HDA19 as bait. A novel protein, bnKCP1, containing a putative kinase-inducible domain (KID) was found to interact with HDA19. Southern blot analysis indicated that the bnKCP1 gene belongs to a small gene family of at least three members. Northern blot analysis showed bnKCP1 to be strongly expressed in stems, flowers, roots, and immature siliques, but not in leaf blades of seedlings. The accumulation of bnKCP1 transcript in the leaf blades was induced significantly within 4 h of exposure of B. napus seedlings to cold stress, whereas treatment of leaf blades with inomycin, an ionophore of Ca2+, caused a rapid (30 min) but transient induction of bnKCP1 expression. In contrast to that observed in leaf blades, expression of bnKCP1 in the stems was repressed upon cold treatment. In vitro and in vivo protein-binding assays showed that bnKCP1 interacts with HDA19 via the KID domain, and that S188 is critical for bnKCP1-HDA19 interaction. BnKCP1 also exerted modest transactivation of the lacZ reporter gene in yeast through its N terminal region. These assays suggest that bnKCP1 may function as a transcription factor, which regulates gene expression through interaction with HDA19. PMID- 12631332 TI - The role of PopCel1 and PopCel2 in poplar leaf growth and cellulose biosynthesis. AB - Poplar calli transcribed two cellulase (endo-1,4-beta-glucanase) genes, PopCel1 and PopCel2, whose mRNAs were differentially located in the growing leaves of poplar during cell wall synthesis. Histochemical and RT-PCR analyses of promoter GUS fusion gene activities in transgenic poplar demonstrated that PopCel1 promoter-derived GUS activity was localized in the petiole and leaf veins, whereas PopCel2 was confined to mesophyll cells and disappeared from the tip during the development of leaves. Autoradiography of the leaf showed that the radioactivity of [14C]sucrose incorporated into cellulose corresponded to the combination of the sucrose-induced tissue-specific patterns of PopCel1 and PopCel2. Interestingly, 2,6-dichlorobenzonitrile (DCB) not only inhibited the incorporation of the radioactivity into cellulose, but also repressed the induction of both cellulase genes. Suppression of cellulases by expression of PopCel1 antisense cDNA or co-suppression of PopCel1 mRNA by overexpression of PopCel1 sense cDNA reduced leaf growth. Therefore, we came to the conclusion that PopCel1 and PopCel2 probably function to promote leaf growth in poplar by the endohydrolysis of 1,4-beta-glucan. PMID- 12631333 TI - Enhancement of growth by expression of poplar cellulase in Arabidopsis thaliana. AB - To study the role of cellulose and cellulase in plant growth, we expressed poplar cellulase (PaPopCel1) constitutively in Arabidopsis thaliana. Expression increased the size of the rosettes due to increased cell size. The change in growth was accompanied by changes in biomechanical properties due to cell wall structure indicative of decrease in xyloglucan cross-linked with cellulose microfibrils by chemical analysis and nuclear magnetic resonance (NMR) spectra. The result supports the concept that the paracrystalline sites of cellulose microfibrils are attacked by poplar cellulase to loosen xyloglucan intercalation and this irreversible wall modification promotes the enlargement of plant cells. PMID- 12631334 TI - Posttransplant erythrocytosis. AB - Posttransplant erythrocytosis (PTE) is defined as a persistently elevated hematocrit to a level greater than 51% after renal transplantation. It occurs in 10% to 15% of graft recipients and usually develops 8 to 24 months after engraftment. Spontaneous remission of established PTE is observed in one fourth of the patients within 2 years from onset, whereas in the remaining three fourths it persists for several years, only to remit after loss of renal function from rejection. Predisposing factors include male gender, retention of native kidneys, smoking, transplant renal artery stenosis, adequate erythropoiesis prior to transplantation, and rejection-free course with well-functioning renal graft. Just as in other forms of erythrocytosis, a substantial number (approximately 60%) of patients with PTE experience malaise, headache, plethora, lethargy, and dizziness. Thromboembolic events occur in 10% to 30% of the cases; 1% to 2% eventually die of associated complications. Posttransplant erythrocytosis results from the combined trophic effect of multiple and interrelated erythropoietic factors. Among them, endogenous erythropoietin appears to play the central role. Persistent erythropoietin secretion from the diseased and chronically ischemic native kidneys does not conform to the normal feedback regulation, thereby establishing a form of "tertiary hypererythropoietinemia." However, erythropoietin levels in most PTE patients still remain within the "normal range," indicating that erythrocytosis finally ensues by the contributory action of additional growth factors on erythroid progenitors, such as angiotensin II, androgens, and insulin-like growth factor 1 (IGF-1). Inactivation of the renin angiotensin system (RAS) by an angiotensin-converting enzyme (ACE) inhibitor, or an angiotensin II type 1 AT1 receptor blocker represents the most effective, safe, and well-tolerated therapeutic modality. PMID- 12631335 TI - Tissue engineering the kidney. AB - The means by which kidney function can be replaced in humans include dialysis and renal allotransplantation. Dialytic therapies are lifesaving, but often poorly tolerated. Transplantation of human kidneys is limited by the availability of donor organs. During the past decades, a number of different approaches have been applied toward tissue engineering the kidney as a means to replace renal function. The goals of one or another of them included the recapitulation of renal filtration, reabsorptive and secretory functions, and replacement of endocrine/metabolic activities. This review will delineate the progress to date recorded for five approaches: (1) integration of new nephrons into the kidney; (2) growing new kidneys in situ; (3) use of stem cells; (4) generation of histocompatible tissues using nuclear transplantation; and (5) bioengineering of an artificial kidney. All five approaches utilize cellular therapy. The first four employ transplantation as well, and the fifth uses dialysis. PMID- 12631336 TI - Polymorphisms in the nephrin gene and diabetic nephropathy in type 1 diabetic patients. AB - BACKGROUND: Several mutations in the nephrin gene are responsible for the lack of slit membrane of the glomeruli leading to massive proteinuria present already in utero. Variations in the nephrin gene may also affect the degree of proteinuria in acquired kidney diseases. We tested the hypothesis of whether any of the polymorphisms identified in the coding region of the nephrin gene were associated with diabetic nephropathy. METHODS: In a case-control, cross-sectional study, 996 Finnish type 1 diabetic patients from the FinnDiane Study were genotyped by standard polymerase chain reaction protocol. RESULTS: The frequencies of the rare alleles in the E117K, R408Q, and N1077S polymorphisms in the entire cohort were 34%, 8%, and 12%, respectively. When comparing patients with a mutant allele with the wild genotype there was no difference between the patients with end-stage renal disease, proteinuria, microalbuminuria, and those with a normal albumin excretion rate (df =3, chi2 =1.62, 1.31 and 0.77). Neither were the polymorphisms associated with the progression of kidney disease, nor with creatinine clearance and albumin excretion rate. CONCLUSION: This study does not support an involvement of the coding region of the nephrin gene in the pathogenesis of diabetic nephropathy in type 1 diabetic patients. PMID- 12631337 TI - Association of an IL-1A 3'UTR polymorphism with end-stage renal disease and IL-1 alpha expression. AB - BACKGROUND: We evaluated polymorphisms in the interleukin-1 alpha 3'-untranslated region (IL-1A 3'[UTR]) for association with type 2 diabetes-associated (DM) and nondiabetic-associated (non-DM) end-stage renal disease (ESRD) in two ethnic groups. METHODS: IL-1A 3'UTR polymorphisms were identified by alignment of overlapping human expressed sequence tags (ESTs). Sequence ambiguities were experimentally confirmed and variants genotyped to test for association with ESRD in 75 unrelated Caucasians with DM ESRD, 95 unrelated Caucasian controls and, in a parallel study, 92 unrelated African Americans with type 2 DM ESRD, 95 unrelated African Americans with non-DM ESRD, and 86 unrelated African American controls. IL-1A 3' UTR genotype and lipopolysaccharide (LPS)-stimulated IL-1 alpha protein levels were measured in healthy Caucasians (N = 112) and African Americans (N = 101) to evaluate association between genotype and protein level. RESULTS: A polymorphism in the 3' UTR of the human IL-1A gene was associated with ESRD and IL-1 alpha protein expression. The polymorphism consists of two single nucleotide polymorphisms (SNPs) and an insertion/deletion generating four different haplotypes: TN7TTCAA, AN7TTCAA, TN7TTCAG and an allele deleted for four internal bases, TN7(delTTCA)A. The 4 bp deletion allele, TN7(delTTCA)A, was significantly less common among Caucasian DM ESRD and African American non-DM ESRD patients (recessive model; P = 0.0364 and P = 0.0293, respectively). In vitro, this polymorphism is associated with the amount of IL-1 alpha protein synthesized in LPS-stimulated lymphocytes from healthy subjects (P = 0.0013, additive model), with the TN7(delTTCA)A haplotype associated with higher levels of stimulated IL-1 alpha. CONCLUSION: The association of the TN7(delTTCA)A haplotype with higher levels of IL-1 alpha expression and reduced risk for ESRD is consistent with involvement of cytokines in risk for developing nephropathy. PMID- 12631338 TI - Delayed cystogenesis and increased ciliogenesis associated with the re-expression of polaris in Tg737 mutant mice. AB - BACKGROUND: Renal cysts and shortened cilia on renal tubular epithelia have been observed in Tg737orpk (orpk) mutant mice, suggesting a potential connection between cystogenesis and ciliogenesis. To further test this hypothesis we have characterized the progression of cystic disease and cilia expression in orpk, orpk;Tg737Rsq (orpk rescue), and Tg737 Delta 2-3 beta Gal;Tg737Rsq (KO rescue) mice. Methods. Orpk, orpk rescue, and KO rescue animals were generated and analyzed from postnatal day (P) 0 to P21 (orpk mutants) or from P0 to P210 (orpk rescue and KO rescue animals). Proximal tubules (PT) and collecting tubules (CT) were identified by immunohistochemistry using segment-specific lectins and a segment-specific cystic index was calculated. Scanning electron microscopy was utilized to observe and measure cilia expression in cysts from orpk, orpk rescue, and KO rescue animals. RESULTS: KO rescue and orpk rescue animals develop adult onset autosomal-recessive polycystic kidney disease (ARPKD). Ultrastructural analysis of cilia expression revealed that cysts from orpk expressed short cilia, whereas cysts from KO rescue animals expressed normal length cilia and cysts from orpk rescue animals expressed cilia that are two to five times longer than wild type. CONCLUSION: While this data is consistent with a role for polaris in ciliogenesis, it does not support a direct connection between ciliogenesis and cystic disease. Similarities in cyst formation and striking differences in cilia expression associated with these ARPKD mouse models indicates that cyst formation and cilia expression are independent phenotypic features regulated by polaris. PMID- 12631339 TI - Evidence that soyasaponin Bb retards disease progression in a murine model of polycystic kidney disease. AB - BACKGROUND: We reported a lessened cyst growth in the pcy mouse model of polycystic kidney disease (PKD) when mice were fed a soy protein isolate (SPI) based diet and hypothesized that the soyasaponins may be associated with this therapeutic effect. The effects of feeding a saponin-enriched alcohol extract (SEAE) from SPI, an isoflavone- and saponin-enriched soy supplement (Novasoy 400), or a 99.5% pure soyasaponin Bb powder on cyst growth are reported here. METHODS: The therapeutic effects of the soyasaponins were studied in 60-day-old male pcy mice in two separate, 90-day feeding trials. In the first study, mice were fed either a casein-based (control) diet, a diet in which SPI replaced the casein or the control diet supplemented with SEAE. In the second study, mice were fed the control diet unsupplemented or supplemented with either a soyasaponin- and isoflavone-enriched soy product (Novasoy 400) or a 99.5% pure soyasaponin Bb powder. RESULTS: In study 1, kidney weight, water content, and plasma creatinine and urea levels were markedly reduced in the SEAE-fed animals compared to tissues from the control group; likewise, mice fed the SPI-based diet showed a decreased plasma creatinine, but only a slightly reduced plasma urea. In study 2, kidney weight, water content, plasma creatinine and urea levels were significantly reduced in mice fed the soyasaponin Bb powder and the Novasoy-400 supplement, compared to controls. CONCLUSION: Soyasaponin Bb can impede kidney enlargement and cyst growth in the pcy mouse model of PKD. Further studies are needed to determine its most effective dose and mechanism of action. PMID- 12631340 TI - Cux-1 transgenic mice develop glomerulosclerosis and interstitial fibrosis. AB - BACKGROUND: Cux-1 is a murine homeobox gene that is highly expressed in the nephrogenic zone of the developing kidney. Transgenic mice ectopically expressing Cux-1 develop renal hyperplasia associated with down-regulation of the cyclin kinase inhibitor p27. Because the reduction of p27 has been associated with mesangial cell proliferation and glomerular disease, we evaluated glomerular changes in Cux-1 transgenic mice. METHODS: Adult kidneys from Cux-1 transgenic mice were analyzed morphologically for changes in glomerular cell number and for changes in mesangial and interstitial extracellular matrix deposition. Mesangial matrix expansion was identified by light microscopy. Glomerular cell number was performed following immunohistochemistry. Type IV collagen deposition was analyzed by immunofluoresence and Western blotting. Renal function was evaluated by serum protein, blood urea nitrogen (BUN), creatinine, and electrolyte analysis, and by urine protein and creatinine analysis. RESULTS: In adult transgenic glomeruli, Cux-1 was ectopically expressed in mesangial cells, and this was associated with an increase in mesangial cell number, resulting from an increase in proliferation. There was a marked increase in mesangial matrix area in transgenic mice compared to non-transgenic littermates, related to an increase in type IV collagen. Podocyte foot process effacement was observed in transgenic mice, and this was related to an increase in urinary albumin. Interstitial fibrosis was also observed in transgenic kidneys. CONCLUSION: These observations indicate that increased expression of Cux-1 in mesangial cells results in cell proliferation and mesangial expansion. In addition, these changes are potentially related to disruption of podocyte architecture leading to loss of filtration. These results suggest that expression of Cux-1 is sufficient to induce the early events of mesangioproliferative glomerulonephritis. PMID- 12631341 TI - IGF-I induces vascular endothelial growth factor in human mesangial cells via a Src-dependent mechanism. AB - BACKGROUND: Both insulin-like growth factor-I (IGF-I) and vascular endothelial growth factor (VEGF) have been implicated in the pathogenesis of early renal dysfunction in diabetes. We investigated whether IGF-I affects VEGF gene expression and protein secretion in human mesangial cells. Furthermore, we studied the intracellular signaling pathway involved and the interaction of IGF-I with mechanical stretch, a known VEGF inducer. METHODS: Human mesangial cells were exposed to IGF-I in the presence and in the absence of (1) anti-IGF-I type I receptor antibody (alpha IR3) (1 microg/mL), a monoclonal antibody blocking the IGF-I type I receptor; (2) wortmannin (600 nmol/L), a phosphatidylinositol 3 kinase (PI3K) inhibitor; (3) 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4 d]pyrimidine (PP2), a specific Src inhibitor (10 micromol/L); and (4) cyclic stretch (approximately 10% elongation). RESULTS: IGF-I induced a dose-dependent increase in VEGF protein levels (10(-11) mol/L, 5%; 10(-10) mol/L, 14%; 10(-9) mol/L, 46%; 10(-8) mol/L, 66%; 10(-7) mol/L, 68%; P < 0.001). IGF-I-induced VEGF production rose by 6 hours with a peak at 12 hours, and declined by 24 hours (52%, 72%, and 34%, respectively; P < 0.01 at 12 hours). A corresponding 50% increase in VEGF mRNA levels was seen at 6 hours (P < 0.01). IGF-I-induced VEGF protein secretion was not affected by the addition of wortmannin (IGF-I, 76% vs. IGF-I + wortmannin, 79% increase over control; P = NS), but was abolished by alpha IR3 (IGF-I, 69% vs. IGF-I +alpha IR3, 0%; P < 0.001) and significantly reduced by PP2 (IGF-I, 50% vs. IGF-I + PP2, 14%; P < 0.01). Simultaneous exposure of human mesangial cells to both IGF-I and stretch failed to further increase VEGF production (IGF-I, 1.49 +/- 0.05; stretch, 1.76 +/- 0.05; and IGF-I + stretch, 1.83 +/- 0.11). CONCLUSION: IGF-I induces VEGF gene expression and protein secretion in human mesangial cells via a Src-dependent mechanism. PMID- 12631342 TI - Activation of the ERK pathway precedes tubular proliferation in the obstructed rat kidney. AB - BACKGROUND: In vitro studies suggest that activation of the extracellular signal regulated kinase (ERK) pathway plays a critical role in the proliferation of tubular epithelial and myofibroblast-like cells. However, little is known of ERK activation in individual cell types in normal or diseased kidney. The aims of this study were to (1) localize ERK activation within the kidney, and (2) examine the relationship between ERK activation and cell proliferation in the injured kidney. METHODS: Unilateral ureteric obstruction (UUO) was induced in groups of six Wistar rats, which were killed at 30 minutes, 6 hours, and 1, 4, or 7 days after obstruction. Activation of ERK was identified using antibodies specific for the phosphorylated form of ERK (pERK) in Western blots and immunostaining. Proliferating cells were detected using bromodeoxyuridine (BrdU). RESULTS: Western blotting showed abundant expression of the two ERK isoforms, ERK-1 and ERK-2, in normal rat kidney. Low levels of activated ERK (pERK-2> pERK-1) were detected in normal rat kidney by Western blotting. Immunostaining showed that ERK activation in normal kidney was largely restricted to collecting ducts in the outer medulla. Within 30 minutes of ureter obstruction, Western blotting showed a sixfold increase in ERK activation followed by a second peak (14-fold increase) on days 4 and 7. The initial peak of ERK activation was localized to medullary collecting ducts and the thick ascending limb of Henle (TALH), whereas the second peak corresponded to a progressive increase in ERK activation in dilated collecting ducts and in interstitial cells in the cortex. Proliferation of tubular epithelial cells closely followed the pattern of ERK activation, being evident first in medullary collecting ducts and TALH on day 1, and then in cortical collecting ducts from day 4. CONCLUSION: This study has identified a discrete pattern of ERK activation in normal rat kidney and an increase in ERK activation following obstruction. The temporal and spatial relationship in which ERK activation preceded tubular cell proliferation suggest that ERK signaling plays a key role in tubular epithelial cell proliferation in the injured kidney. PMID- 12631343 TI - Induction of MIF synthesis and secretion by tubular epithelial cells: a novel action of angiotensin II. AB - BACKGROUND: Angiotensin II (Ang II) plays an important role in the development of renal injury through its vasoactive and proinflammatory activities. We investigated whether some of the effects of Ang II could be mediated through the production of macrophage migration inhibitory factor (MIF). METHODS: Groups of rats underwent sham surgery (sham), subtotal nephrectomy (STNx), or STNx plus treatment with irbesartan. Renal tissue was examined 12 weeks postsurgery for MIF mRNA expression and leukocyte accumulation. To determine whether Ang II had a direct effect on MIF production, mRNA synthesis and protein secretion were examined in proximal tubular epithelial (NRK52E and MCT) cell lines. RESULTS: MIF mRNA was strongly expressed in 5.4%+/- 1.1% (mean +/- SD) of cortical tubules of sham-operated rats. This was significantly up-regulated in STNx rats (44.9%+/- 22.6%) and was abrogated by administration of irbesartan (2.8%+/- 2.4%). STNx resulted in significant glomerular and interstitial accumulation of macrophages and T cells, which correlated with glomerular and tubular MIF mRNA expression, respectively. In vitro studies of tubular epithelial cells revealed that Ang II caused a twofold increase in MIF mRNA expression in NRK52E and MCT cells, which was abrogated by irbesartan. In addition, Ang II induced a rapid release of 50% of MIF protein from NRK52E cells within 20 minutes. CONCLUSION: This study has demonstrated that Ang II up-regulates MIF mRNA production and MIF protein secretion by tubular epithelial cells. Ang II may promote accumulation and activation of interstitial leukocytes via induction of MIF synthesis and secretion in renal tubular epithelial cells. This may be an important mechanism by which Ang II mediates renal injury. PMID- 12631345 TI - Altered polarity and expression of H+-ATPase without ultrastructural changes in kidneys of Dent's disease patients. AB - BACKGROUND: Dent's disease is a proximal tubule (PT) disorder characterized by low-molecular-weight proteinuria (LWMP) that may be associated with hypercalciuria, nephrocalcinosis, and renal failure. It is caused by inactivating mutations of the renal chloride channel ClC-5, which colocalizes with the vacuolar H+-ATPase in PT cells and alpha-type intercalated cells. Examinations of knockout mice have established the role of ClC-5 in PT endocytosis, but the consequences of ClC-5 mutations on the polarity of H+-ATPase and other plasma membrane proteins remain unknown. METHODS: We have studied renal biopsies from eight patients with Dent's disease, due to inactivating ClC-5 mutations, by light and electron microscopy, and by immunohistochemical staining. All patients exhibited LMWP, and renal function ranged from normal to end-stage renal failure. RESULTS: Light microscopy revealed either normal renal architecture or glomerulosclerosis, tubular dedifferentiation and atrophy, and mild interstitial fibrosis. Focal, hyaline casts, sometimes calcified, were identified at all stages. Electron microscopy did not reveal any ultrastructural abnormalities in PT cells, and the endocytic apparatus was apparently normal. However, immunohistochemical studies demonstrated a consistent inversion of H+-ATPase polarity in PT cells to a basolateral distribution contrasting with its apical location in the normal kidney. This inversion of polarity was specific for H+ ATPase and did not affect distribution of aminopeptidase, megalin, and Na+/K+ ATPase. Furthermore, apical H+-ATPase expression was absent in alpha-type intercalated cells. CONCLUSION: ClC-5 mutations are associated with modifications in the polarity and expression of H+-ATPase, but not ultrastructural alterations in PT cells. These findings help further understanding of the role of ClC-5 and the pathophysiology of Dent's disease. PMID- 12631344 TI - ETA receptor-mediated Ca2+ signaling in thin descending limbs of Henle's loop: impairment in genetic hypertension. AB - BACKGROUND: Endothelins (ET) have diuretic and natriuretic actions via ETB receptors that are found in most renal tubular segments, although the thin limbs have not been studied. Data also suggest that dysfunction of the renal ET system may be important in the pathogenesis of hypertension. The present study was aimed at determining the presence and nature of ET receptors in the thin limbs of Henle's loop and their ability to activate a Ca2+-dependent signaling pathway, as well as whether ET-induced Ca2+ signals are altered in hypertension. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and Fura 2 fluoreselected strains of Lyon rats with low-normal (LL), normal (LN), and high (LH) blood pressure. RESULTS: In SD rats, ET induced Ca2+ signals in DTL of long looped nephrons, but not in DTL of short loops, or in ascending thin limbs. Ca2+ increases were abolished by BQ123, an antagonist of the ETA receptor, but not by BQ788, an antagonist of the ETB subtype. Endothelin-3 and sarafotoxin 6c, two ETB receptor agonists, were both inactive. RT-PCR showed the presence of both ETA and ETB receptor mRNA. Ca2+ signals measured scence measurements of [Ca2+]i were made to characterize ET receptors in descending thin limbs (DTL) of Sprague-Dawley rats, spontaneously hypertensive (SH) rats, and control Wistar-Kyoto (WKY) rats, and the three in DTL of WKY LL and LN rats were similar to those in Sprague Dawley rats, but were significantly diminished (LH) or abolished (SH) in hypertensive rats. CONCLUSION: A functional ETA receptor activating a Ca2+ dependent pathway is expressed in DTL. This ETA-induced calcium signaling is impaired in two strains of genetically hypertensive rats. PMID- 12631346 TI - Hyperphosphatemia aggravates cardiac fibrosis and microvascular disease in experimental uremia. AB - BACKGROUND: Hyperphosphatemia is a known predictor of cardiovascular death and specifically of cardiac death in hemodialysis patients. The pathomechanisms involved have not been completely clarified. While a number of observations suggest an important role of hyperphosphatemia and positive calcium balance on atherosclerosis and calcification of the coronary conduit arteries, independent effects on postcoronary microvessels and on cardiac fibrosis have not been excluded. METHODS: Male Sprague-Dawley rats were sham operated (N = 14) or subtotally nephrectomized (SNX, N = 17) and subsequently placed on low phosphorus (0.08% w/w) and high phosphorus (1.2% w/w) diet under pair-feeding conditions. After 8 weeks, serum chemistry and inhibitory parathyroid hormone (iPTH) were measured, and the hearts were harvested using perfusion fixation. Arteriolar thickness and volume density of the interstitium (excluding vessels) were quantitated using stereologic techniques. RESULTS: In SNX animals with moderate renal failure serum phosphorus concentrations were higher than in sham-operated controls on low phosphorus diet (1.7 +/- 0.37 mmol/L) and were significantly higher in SNX + high phosphorus diet (2.33 +/- 0.23 mmol/L) compared to SNX + low phosphorus diet (1.95 +/- 0.32 mmol/L; P < 0.05). In sham-operated controls, dietary phosphorus content had no effect on cardiac morphologic indices. In contrast, in SNX + high phosphorus diet the index of interstitial cardiac fibrosis was significantly higher (3.22 +/- 0.44%) than in SNX + low phosphorus (2.75 +/- 0.46%) or in sham-operated controls (2.5 +/- 0.05% on high phosphorus and 2.4 +/- 0.89 on low phosphorus, respectively). In SNX + high phosphorus (14.0 +/- 9.0 microm), but not in SNX + low phosphorus (9.2 +/- 4.5 microm), arterial wall thickness was significantly higher compared to sham-operated controls (10.2 +/- 5.1 on high phosphorus and 9.8 +/- 5.0 micro;m on low phosphorus, respectively). The data were confirmed in an independent repeat experiment. CONCLUSION: High dietary phosphorus and hyperphosphatemia have significant effects on cardiac fibrosis and arterial wall thickening. Such abnormalities of cardiac architecture may be relevant for the increased cardiac risk in hyperphosphatemic uremic patients. PMID- 12631347 TI - Specific inhibition of Egr-1 prevents mesangial cell hypercellularity in experimental nephritis. AB - BACKGROUND: Mesangial cell proliferation is a frequent finding in glomerulonephritis. In cultured mesangial cells, we demonstrated that inhibition of the zinc finger transcription factor, early growth response gene-1 (Egr-1), by specific antisense oligonucleotides (AS ODN) blocks mesangial cell proliferation. Therefore, we here investigated the effect of Egr-1 inhibition on the course of an experimental mesangioproliferative glomerulonephritis in vivo. METHODS: On day 3 after induction of anti-Thy-1.1 nephritis, specific glomerular oligonucleotide transfer was achieved by injection of an oligonucleotide/hemagglutinating virus of Japan/liposome mixture into the left renal artery. The right kidney was left untreated. RESULTS: Induction of nephritis led to a sixfold induction of Egr-1 protein on day 6 of disease. This increase in Egr-1 expression was reduced by 48% in the left kidney by transfer of specific AS ODN. In parallel, the increases in glomerular cellularity, number of mitoses, and glomerular tuft area observed in day 6 nephritic animals were inhibited in the left kidney by 60%, 53%, and 50%, respectively. Changes in the right kidney were not significantly influenced. Likewise, control oligonucleotides showed no effect. Finally, the expression of platelet-derived growth factor-B (PDGF-B), a known target gene of Egr-1, was repressed by transfer of specific AS ODN against Egr-1. CONCLUSION: We conclude that the transcription factor Egr-1 plays a critical role for mesangial cell proliferation in vivo. Interfering with the induction of Egr-1 or with its target genes could give rise to novel therapeutic principles in mesangioproliferative glomerulonephritis. PMID- 12631348 TI - Crystal attachment to injured renal collecting duct cells: influence of urine proteins and pH. AB - BACKGROUND: The attachment of crystals to injured kidney epithelium is thought to be a necessary event in the development of urolithiasis. In vivo, the crystals are coated with urinary macromolecules that define the surface properties of the crystals. The present study examines the influence of coating of calcium oxalate crystals with urinary macromolecules on their attachment to both healthy (polarized) and injured (nonpolarized) primary inner medullary collecting duct (IMCD) cells. METHODS: Calcium oxalate monohydrate (COM) and calcium oxalate dihydrate (COD) crystals were coated with urine macromolecules by incubating the crystals in urine from normal healthy volunteers at pH 5, 6, and 7. The level of attachment of the coated crystals to IMCD cells was also determined at pH 5, 6, and 7. The adsorbed proteins were extracted from the crystal surfaces and separated by gel electrophoresis. RESULTS: The coating of calcium oxalate crystals with urine proteins greatly reduced the attachment of crystals to both control and injured IMCD cells. At pH levels below 6, the crystals readily attached to injured cells. Extraction and separation of the adsorbed proteins showed that both COM and COD crystals adsorbed a similar array of proteins. At pH 5 and 6, several trace proteins were adsorbed to the crystals and were not apparent at pH 7. CONCLUSION: The coating of crystals with urine macromolecules greatly reduces the attachment of the crystals to normal healthy epithelia. The coating and attachment of the crystals below pH 6 enhances the attachment to injured cells. The enhanced crystal attachment could possibly be associated with one or more proteins adsorbed to the crystal surface that are not adsorbed to the crystals at higher pH. PMID- 12631349 TI - Distinct distribution of specific members of protein 4.1 gene family in the mouse nephron. AB - BACKGROUND: Protein 4.1 is an adapter protein that links the actin cytoskeleton to various transmembrane proteins. These 4.1 proteins are encoded by four homologous genes, 4.1R, 4.1G, 4.1N, and 4.1B, which undergo complex alternative splicing. Here we performed a detailed characterization of the expression of specific 4.1 proteins in the mouse nephron. METHODS: Distribution of renal 4.1 proteins was investigated by staining of paraformaldehyde-fixed mouse kidney sections with antibodies highly specific for each 4.1 protein. Major 4.1 splice forms, amplified from mouse kidney marathon cDNA, were expressed in transfected COS-7 cells in order to assign species of known exon composition to proteins detected in kidney. RESULTS: A 105 kD 4.1R splice form, initiating at ATG-2 translation initiation site and lacking exon 16, but including exon 17B, was restricted to thick ascending limb of Henle's loop. A 95 kD 4.1N splice form, lacking exons 15 and 17D, was expressed in either descending or ascending thin limb of Henle's loop, distal convoluted tubule, and all regions of the collecting duct system. A major 108 kD 4.1B splice form, initiating at a newly characterized ATG translation initiation site, and lacking exons 15, 17B, and 21, was present only in Bowman's capsule and proximal convoluted tubule (PCT). There was no expression of 4.1G in kidney. CONCLUSION: Distinct distribution of 4.1 proteins along the nephron suggests their involvement in targeting of selected transmembrane proteins in kidney epithelium and, therefore, in regulation of specific kidney functions. PMID- 12631350 TI - Monocytes may promote myofibroblast accumulation and apoptosis in Alport renal fibrosis. AB - BACKGROUND: In interstitial fibrosis, monocytes and myofibroblasts have been directly implicated in scarring, apoptosis, and tissue necrosis. While much has been done to explore the role of these cell types individually in fibrosis, the interactive dependency of monocytes and myofibroblasts has been only marginally explored. METHODS: Alport mice were treated or not with a soluble receptor inhibitor for transforming growth factor-beta 1 (TGF-beta 1), which was previously shown to inhibit the accumulation of myofibroblasts, but not monocytes, in the tubulointerstitium. Kidneys were examined for fibrosis using several matrix markers, TGF-beta 1 mRNA expression by in situ hybridization, apoptosis using the terminal deoxynucleotidyl transferase-mediated uridine triphosphate nick end labeling (TUNEL) assay, expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPS) by dual immunofluorescence microscopy, MMP activity by gelatin and in situ zymography, MMP mRNA expression by reverse transcription-polymerase chain reaction (RT-PCR), and basement membrane degradation by dual immunofluorescence confocal microscopy and electron microscopy. RESULTS: Treated mice showed a markedly reduced accumulation of matrix proteins. Tissue monocytes express TGF beta 1 mRNA, and TGF-beta 1 is required for myofibroblast accumulation. The number of apoptotic cells was not influenced by TGF-beta 1 inhibition. Monocytes express MMP-2, MMP-9, TIMP-2, and TIMP-3. MMP activity and mRNA expression is equally up regulated in treated and untreated Alport mice. Tubular basement membranes (TBM) around clusters of monocytes are notably degraded. TGF-beta 1 inhibition does not extend the life of Alport mice. CONCLUSION: These studies demonstrate that monocytes may influence myofibroblast accumulation via TGF beta1, and that monocytes, and not myofibroblasts, are associated with tubular atrophy in Alport mice. Elevated MMP expression and activity is associated with TBM destruction near monocytes clusters, suggesting an anoikis mechanism may contribute to apoptosis in this model. PMID- 12631351 TI - Melatonin induced cyclic modulation of vectorial water transport in kidney derived MDCK cells. AB - BACKGROUND: Melatonin, newly synthesized by the pineal gland, is rapidly released to general circulation reaching a nanomolar concentration. Cyclic production of melatonin synchronizes body rhythms with the photoperiod. Moreover, changes in urine production and osmolarity have been observed in the kidney during the night. However, the precise mechanisms by which plasma-circulating melatonin modifies renal physiology are not clearly understood. METHODS: Madin-Darby canine kidney (MDCK) cell monolayers transport water vectorially from the apical to the basolateral side forming blisters or domes. Transport in epithelial cells is regulated by tight junction sealing, ion pumps and channels, and cytoskeleton organization, among other processes. MDCK cells were used to study vectorial water transport to determine the role of microfilament organization and protein kinase C (PKC) in dome formation in culture conditions that mimic the cyclic pattern of melatonin circulation in plasma. RESULTS: Melatonin cyclically increased dome formation by 50% and caused enlargement and thickening of stress fibers in cells surrounding the domes. Optimal increase in dome formation was observed at nanomolar concentrations of melatonin after 6 hours, concomitantly with a 28% decrease in the transepithelial electrical resistance, which remained low for up to 12 hours, without apparent change in fluorescein isothiocyanate (FITC)-dextran flux. A blockage in dome formation elicited by melatonin was observed in monolayers preincubated with the Na+-K+-ATPase or PKC inhibitors. CONCLUSION: The results obtained indicate that melatonin cyclically modifies the transepithelial permeability in kidney-derived cells through PKC activation and microfilament reorganization, and supports the hypothesis that melatonin may synchronize daily body rhythms through cyclic cytoskeletal rearrangements. PMID- 12631352 TI - Intussusceptive capillary growth is required for glomerular repair in rat Thy-1.1 nephritis. AB - BACKGROUND: Injection of anti-Thy-1.1 antibodies to rats causes mesangiolysis, with subsequent capillary loss. This dramatic event is followed by almost complete recovery of glomerular architecture. However, the precise cellular mechanisms of revascularization are not fully understood. METHODS: Glomerulonephritis was induced by the injection of monoclonal anti-Thy-1.1 antibody to rats. Structural changes in the glomerular vasculature, with special emphasis on the repair phase, were studied with corrosion casting technique, light microscopy (LM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). RESULTS: Specifically during the extensive revascularization period, numerous tiny holes of about 1.5 microm in diameter were found on the glomerular cast. The presence of transluminal tissue pillar was confirmed with LM and SEM. These findings indicate the involvement of intussusceptive capillary growth. TEM study demonstrated that some tissue pillars were composed of endothelial cells only and others had cytoplasmic process of mesangial cells in their cores sandwiched both sides by endothelial walls. CONCLUSION: Intussusceptive capillary growth, or nonsprouting angiogenesis is involved in the postinjury angiogenesis in the glomeruli, in which process mesangial cells as well as endothelial cells may play important roles. PMID- 12631353 TI - Cell-cycle regulatory proteins in podocyte cell in idiopathic nephrotic syndrome of childhood. AB - BACKGROUND: The podocyte cell is believed to play an important role in idiopathic nephrotic syndrome (INS) of childhood. In adults with cellular and collapsing focal segmental glomerulosclerosis (FSGS), the expression of cell-cycle regulatory proteins such as p27, p57, and cyclin D is decreased and expression of cyclin A, Ki-67, and p21 is observed in podocyte cells suggestive of a dysregulated podocyte phenotype. We investigated for alterations in the expression of cyclin kinase inhibitors, p27, p57, p21, and cyclins D and A in the podocyte cell of children with INS. METHODS: Forty-two kidney biopsies were investigated; 14 with minimal-change disease (MCD), seven with diffuse mesangial hypercellularity (DMH), 12 with FSGS, four with Alport syndrome (AS), and five normal biopsies. The sections were examined by immunohistochemistry using dual staining method. Podocyte cells were first identified by Wilm's tumor-1 staining after which expressions of cell-cycle regulatory proteins were analyzed. A quantitative analysis was performed for the proportion of podocyte cells that expressed each cell cycle regulatory protein. RESULTS: On light microscopy, all podocyte cells expressed p27, while p57 and p21 expression was seen in a portion of podocyte cells in normal kidney biopsies. Cyclin D was expressed in a small percent of podocyte cells though the expression was more marked in mesangial and endothelial cells. Cyclin A expression was not seen in normal biopsies. The mean expression of p27 decreased significantly in order from normal (100%), MCD (45.9%), DMH (22.4%), and FSGS (16.7%), and the difference between MCD and FSGS was significant. p21 was significantly and equally reduced in MCD (2.3%), DMH (0%), and FSGS (0.7%) compared to normal (66.6%). There was no significant difference in expression of p57, cyclin D and cyclin A in the podocyte cells between normal and children with INS. Children with AS showed a significant decrease in p27 and p21 expression, while the expression of p57, cyclin D and cyclin A were unchanged from normal, thus demonstrating a pattern similar to INS. CONCLUSION: The podocyte cell in children with INS down-regulates expression of cyclin kinase inhibitors such as p21 and p27, but not p57, but does not up regulate cyclin D and cyclin A that are needed to overcome the G1/S transition and move the cell forward in the cell cycle process. Thus, the podocyte cell remains trapped in the G1 arrest phase. In children with INS or AS, the dysregulated podocyte phenotype is different than the one described in adults with cellular or collapsing FSGS. PMID- 12631354 TI - L-arginine supplementation accelerates renal fibrosis and shortens life span in experimental lupus nephritis. AB - BACKGROUND: Inducible, high-output nitric oxide (NO) production has been identified as a central mediator of cell injury in immune-mediated renal disease. In acute anti-thy-1 glomerulonephritis prefeeding with the NO precursor L arginine increases mesangial cell injury and the subsequent fibrosis. The present study tested the hypothesis that L-arginine supplementation may also be detrimental in chronic, NO-mediated murine lupus nephritis. METHODS: Groups (N = 18) of female MRL/lpr mice with lupus nephritis were fed the following diets: (1) normal protein (22% casein); (2) normal protein and 1.0% L-arginine in the drinking water; (3) low protein (6% casein); (4) low protein + 0.4%l-arginine; and (5) low protein + 1.0% L-arginine. After 40 days mouse survival, albuminuria, matrix accumulation, inflammatory cell infiltration, immunoglobulin G (IgG) deposition, expression of transforming growth factor-beta 1 (TGF-beta 1), fibronectin and plasminogen activator inhibitor-1 (PAI-1) mRNA and protein, anti DNA antibody titer, inducible nitric oxide synthase (iNOS) mRNA expression, blood amino acid levels, blood urea nitrogen (BUN) concentrations and blood and urinary NOx (nitrite + nitrate) levels were assessed. RESULTS: L-Arginine supplementation increased mortality significantly (P < 0.02). The death rate increased from 0% in the lowest to 50% in the highest L-arginine intake group (normal protein + 1.0% L arginine). L-Arginine administration increased albuminuria, renal matrix accumulation, TGF-beta 1, fibronectin, PAI-1, blood L-arginine, L-citrulline, BUN and blood and urine NOx levels, while protein restriction reduced these parameters. Renal cell infiltration and iNOS mRNA expression were decreased in the low protein group only. Anti-ds DNA-IgG and renal IgG deposition were comparable in all groups CONCLUSIONS: Increasing L-arginine intake increases the severity of renal fibrosis and the likelihood of death in MRL/lpr mice. The results appear to be at least in part mediated through enhanced cytotoxic NO generation via iNOS. The data suggest that L-arginine restriction should be considered in human immune-mediated renal diseases. PMID- 12631355 TI - Preconditioning with tin-protoporphyrin IX attenuates ischemia/reperfusion injury in the rat kidney. AB - BACKGROUND: Heme oxygenase (HO)-1 is induced as a unique stress response and leads to a transient resistance against oxidative damage, including ischemia and reperfusion (I/R) injury. In the present study, we examined whether HO-1 induction may confer a protection against I/R injury in the rat kidney. METHODS: Lewis rats were divided into four groups as follows: (1) vehicle group; (2) group treated with ferri-protoporphyrin IX (hemin), an inducer of HO; (3) group treated with low-dose tin-protoporphyrin IX (SnPP), an inhibitor of HO; and (4) group treated with high-dose SnPP. Renal warm ischemia for 60 minutes was performed 24 hours after each treatment. RESULTS: At 24 hours after treatment, hemin induced a significant increase in renal HO activity, but failed to induce HO-1 protein synthesis. Although both low- and high-dose SnPP reduced HO activity, a marked HO 1 expression was observed only in the high-dose SnPP-treated kidney. Hemin exacerbated the renal function after reperfusion, while high-dose SnPP significantly suppressed the intercellular adhesion molecule (ICAM)-1 expression, the infiltration of ED-1-positive macrophages and the expression of activated caspase-3, which resulted in attenuation of apoptotic cell death and ameliorated I/R injury. CONCLUSION: These results suggest that prior induction of HO-1 protein by high-dose SnPP may lead to anti-inflammatory and antiapoptotic effects on warm renal I/R injury independently of its enzyme activity, and that HO enzyme activation may not always act as an antioxidant, especially under I/R-induced oxidative stress. PMID- 12631356 TI - Glucose-mediated induction of TGF-beta 1 and MCP-1 in mesothelial cells in vitro is osmolality and polyol pathway dependent. AB - BACKGROUND: Glucose is converted to sorbitol and then to fructose via the polyol pathway that has been implicated in the pathogenesis of organ damage. The contribution of the polyol pathway to mesothelial cell activation has, however, not been fully determined. METHODS: The effect of increasing glucose concentrations on transforming growth factor-beta 1 (TGF-beta 1) and monocyte chemoattractant protein-1 (MCP-1) secretion by human peritoneal mesothelial cells (HPMC) was examined. The importance of the polyol pathway was identified by its specific inhibition with an aldose reductase inhibitor. RESULTS: Incubation of HPMC with 5 to 100 mmol/L glucose resulted in an induction of aldose reductase mRNA and intracellular sorbitol accumulation accompanied by the induction of TGF beta 1 and MCP-1 mRNA expression and protein secretion. Mannitol at the same concentrations also induced aldose reductase, TGF-beta 1 and MCP-1 mRNA and protein expression but at a lower level than glucose. Sorbinil dose-dependently reduced both intracellular sorbitol levels (79.8% reduction of 60 mmol/L D glucose induced intracellular sorbitol with 100 micromol/L sorbinil (N = 3, P < 0.01) and glucose-induced TGF-beta 1 and MCP-1 secretion. Mannitol induced TGF beta 1 and MCP-1 secretion was not reduced by sorbinil. The addition of 15 to 40 mmol/L sodium lactate, either alone or in the presence of D-glucose enhanced TGF beta 1 and MCP-1 secretion, which was inhibited by sorbinil. In contrast, sodium pyruvate appeared to antagonize D-glucose-induced TGF-beta 1 and MCP-1 secretion. CONCLUSION: These data suggest that the polyol pathway and osmolality contribute to the regulation of HPMC function by glucose. Control of polyol pathway activation might reduce glucose-mediated damage to the peritoneal membrane and promote its long-term survival. PMID- 12631357 TI - Urinary aquaporin-2 in healthy humans and patients with liver cirrhosis and chronic heart failure during baseline conditions and after acute water load. AB - BACKGROUND: Patients with liver cirrhosis and chronic heart failure (CHF) have a reduced capacity to excrete water. Studies in healthy humans have shown that an acute water load reduces the excretion of aquaporin-2 in urine (u-AQP-2). We wanted to test the hypothesis that an acute water load reduces u-AQP-2 less in patients with liver cirrhosis or CHF than in healthy humans. METHODS: Fourteen healthy subjects, 14 patients with liver cirrhosis, and 14 patients with CHF were given an oral water load of 20 mL/kg. Urine was collected every 30 minutes for 4 hours for analysis of u-AQP-2. Blood samples were drawn at the beginning and at the end of the study for analysis of arginine vasopressin (AVP). u-AQP-2 was determined by radioimmunoassay. RESULTS: During the study period, urinary output was 22.8% higher than water intake in the healthy controls and increased 14-fold from baseline, but in patients with liver cirrhosis and CHF urinary output was 14% and 24% less than the intake, while urinary output increased 7- and 19-fold from baseline, respectively. u-AQP2 decreased significantly more in patients with CHF (39%) than in healthy controls (17%) but it was unchanged in those with liver cirrhosis. AVP decreased 46% in patients with CHF, but was unchanged in healthy controls and those with liver cirrhosis. A 24-hour urinary excretion of AQP-2 was significantly elevated in patients with CHF (median, 25.7 nmol/mol creatinine) compared to healthy controls (15.7 nmol/mol creatinine) and those with liver cirrhosis (17 nmol/mol creatinine). CONCLUSION: The excretion of AQP-2 in urine is abnormal both in liver cirrhosis in which we find less suppression of u-AQP2 by an acute water load and in CHF in which we find a high baseline level and an exaggerated suppression of u-AQP2 by an acute water load. PMID- 12631358 TI - Modeling study of human renal chloride channel (hCLC-5) mutations suggests a structural-functional relationship. AB - BACKGROUND: Dent's disease, a renal tubular disorder characterized by low molecular-weight proteinuria, hypercalciuria, and nephrolithiasis, is due to inactivating mutations in the X-linked renal-specific chloride channel, hCLC-5. The x-ray crystal structures of two bacterial chloride channels (CLCs) have recently been established, thereby allowing us to construct a model for hCLC-5 and further examine the role of its mutations. METHODS: The data regarding 49 hCLC-5 mutations were reviewed. Thirty-four mutations that predicted absent or truncated channels were excluded. The remaining 15 mutations (one in-frame insertion and 14 missense mutations), 12 of which have been studied electrophysiologically, were assessed. The hCLC-5 sequence was aligned with the Salmonella typhimurium and Escherichia coli sequences and used to map the hCLC-5 mutations onto a three-dimensional model. RESULTS: hCLC-5 is a homodimeric protein, with each subunit consisting of 18 helices. None of the missense mutations involved the chloride (Cl-) selectivity filter, but 12 of the 15 mutations were found to be clustered at the interface of the two subunits. Six of these mutations occurred in two of the helices that either form part of the interface or lie in close proximity to the interface, and three other mutations that did not lead to complete loss of Cl- conductance were at the edge of the interface. CONCLUSION: These results demonstrate a crucial role for the interaction between the two subunits at the interface of the homodimeric hCLC-5. PMID- 12631359 TI - Vitamin C improves resistance but not conduit artery endothelial function in patients with chronic renal failure. AB - BACKGROUND: Chronic renal failure is associated with impaired endothelium dependent vasodilation and accelerated atherogenesis. To examine whether endogenous reactive oxygen species (ROS) modify endothelial function in renal failure, we evaluated the effect of the antioxidant vitamin C on endothelium dependent responses in both the conduit and resistance vasculature of subjects with severe renal impairment. METHODS: Endothelial function of the forearm resistance vasculature was assessed using plethysmography to measure the dilator response to intra-arterial acetylcholine (Ach) (25 to 100 nmol/min). Endothelial function of radial and brachial arteries was assessed using vascular ultrasound to measure the dilator response to flow during reactive hyperemia [flow-mediated dilatation (FMD)]. Studies were performed before and after administration of vitamin C by intra-arterial infusion (25 mg/min) in 33 predialysis patients or by intravenous infusion (3 g) in 17 hemodialysis patients. RESULTS: Parenteral administration of vitamin C resulted in a 100-fold increase (intra-arterial studies) and a 4.5-fold increase (intravenous studies) in serum antioxidant activity. Vitamin C administration increased the dilator response to ACh in resistance vessels (P = 0.01), but did not alter the dilator response to flow in conduit vessels of either dialysis (P = 0.3) or predialysis subjects (P = 0.8). In the presence of the nitric oxide (NO) synthase inhibitor NGmonomethyl-L arginine (L-NMMA), there was no effect of vitamin C on resistance vessel endothelial function. In all cases the dilator response to the endothelium independent dilators was unaffected by vitamin C. CONCLUSION: Acute administration of vitamin C reduces oxidant stress in renal failure and improves NO-mediated resistance vessel dilatation. PMID- 12631360 TI - Interactions between gender and the angiotensin type 1 receptor gene polymorphism. AB - BACKGROUND: The cardiovascular effects of angiotensin II are mediated by the angiotensin II type 1 receptor (AGT1R); one polymorphism of the AGT1R gene, A1166 ->C, has been associated with hypertension. The hemodynamic response to angiotensin II is blunted in women compared to men, but interactions between gender, blood pressure, and AGT1R gene polymorphisms are unclear. METHODS: A total of 81 young healthy normotensive individuals maintained regulated sodium and protein intake prior to study. They were divided into four groups based on gender and A1166-->C genotype (AA versus AC/CC); serial supine blood pressures were obtained. A subset of 52 individuals received graded infusions of angiotensin II. Inulin and paraaminohippurate clearance techniques were used to measure renal hemodynamic function at baseline and in response to the infusions. RESULTS: Men with the AC/CC genotype exhibited higher blood pressures than men with the AA genotype; however, this relationship was not found among women. Analysis of covariance revealed a significant interaction between gender and AGT1R genotype in the determination of blood pressure. Glomerular filtration rate (GFR) declined variably in the study subjects following infusion of angiotensin II, and a statistical model incorporating gender and genotype best predicted the fall in GFR. There was a trend for females of the AA genotype to have a greater fall in GFR in response to angiotensin II infusion, than any of the other groups. CONCLUSION: In young healthy subjects, there is an important interaction between gender, the AGT1R A1166-->C gene polymorphism, and blood pressure. In addition, the renal hemodynamic response to angiotensin II infusion is a function of both gender and the AGT1R genotype. PMID- 12631361 TI - Fibrillary and immunotactoid glomerulonephritis: Distinct entities with different clinical and pathologic features. AB - BACKGROUND: Controversy surrounds the relatedness of fibrillary glomerulonephritis (FGN) and immunotactoid glomerulonephritis (IT). METHODS: To better define their clinicopathologic features and outcome, we report the largest single center series of 67 cases biopsied from 1980 to 2001, including 61 FGN and 6 IT. FGN was defined by glomerular immune deposition of Congo red-negative randomly oriented fibrils of < 30 nm (mean, 20.1 +/- 0.4 nm). IT was defined by glomerular deposition of hollow, stacked microtubules of > or = 30 nm (mean, 38.2 +/- 5.7 nm). RESULTS: FGN comprised 0.6% of total native kidney biopsies and IT was tenfold more rare (0.06%). Deposits in FGN were immunoglobulin G (IgG) dominant and polyclonal in 96%. IgG subtype analysis in 19 FGN cases showed monotypic deposits in four (two IgG1 and two IgG4) and oligotypic deposits in 15 (all combined IgG1 and IgG4). In IT, deposits were IgG dominant in 83% and monoclonal in 67% (three IgG1 kappa and one IgG1 lambda). FGN patients were a mean age of 57 years, 92% were Caucasian, and 39% were male. At biopsy, FGN patients had the following clinical characteristics (mean, range): creatinine 3.1 mg/dL (0.5 to 14), proteinuria 6.5 g/day (0.8 to 25), 60% microhematuria, and 59% hypertension. Histologic patterns of FGN were diverse, including diffuse proliferative glomerulonephritis (DPGN) (nine cases), membranoproliferative glomerulonephritis (MPGN) (27 cases), mesangial proliferative/sclerosing (MES) (13), membranous glomerulonephritis (MGN) (four), and diffuse sclerosing (DS) (eight). The more proliferative (MPGN and DPGN) and sclerosing (DS) forms presented with a higher creatinine and greater proteinuria compared to MES and MGN. Median time to end-stage renal disease (ESRD) was 24.4 months for FGN and mean time to ESRD varied by histologic subtype: DS 7 months, DPGN 20 months, MPGN 44 months, compared to MES 80 months and MGN 87 months. There was no statistically significant effect of immunosuppressive therapy (given to 36% of FGN patients). By Cox regression (hazard ratio, confidence interval, P value), independent predictors of progression to ESRD were creatinine at biopsy [2.05 (1.55 to 2.72) P < 0.001] and severity of interstitial fibrosis [2.01 (1.05 to 3.85) P = 0.034]. Although IT had similar presentation, histologic patterns, and outcome compared to FGN, it had a greater association with monoclonal gammopathy (P = 0.014), underlying lymphoproliferative disease (P = 0.020), and hypocomplementemia (P = 0.032). CONCLUSION: FGN is an idiopathic condition characterized by polyclonal immune deposits with restricted gamma isotypes. Most patients present with significant renal insufficiency and have a poor outcome despite immunosuppressive therapy, and outcome correlates with histologic subtype. By contrast, IT often contains monoclonal IgG deposits and has a significant association with underlying dysproteinemia and hypocomplementemia. Differentiation of FGN from the much more rare entity IT appears justified on immunopathologic, ultrastructural, and clinical grounds. PMID- 12631362 TI - Smoking and cardiovascular outcomes in dialysis patients: the United States Renal Data System Wave 2 study. AB - BACKGROUND: Smoking has received surprisingly little research attention in dialysis populations, a group at monumental cardiovascular risk. METHODS: Medicare claims data were used to study associations between smoking and new onset cardiovascular outcomes, and death in the prospective, inception Wave 2 cohort (N = 4024), assembled in 1996 and 1997. RESULTS: Of the participants, 56.4% were lifetime nonsmokers, 3.6% were smokers with unknown current status, 20.0% had quit for more than 1 year, 5.8% had quit less than 1 year ago, and 14.2% were current smokers. Subjects with cardiovascular disease at baseline were more likely to be former smokers, less likely never to have smoked and less likely to be current smokers (P < 0.001). Patients were followed until December 31, 1998. When adjustment was made for baseline age, demographic variables, mode of dialysis therapy, and comorbidity, smoking status was associated with new onset congestive heart failure (adjusted hazards ratio 1.59 comparing current to nonsmokers, P = 0.004), new-onset peripheral vascular disease (adjusted hazards ratio 1.68, P < 0.001), and mortality (adjusted hazards ratio 1.37, P < 0.001). Former smokers, in contrast, had adjusted event risks similar to lifelong nonsmokers. CONCLUSION: Smoking is a major, modifiable, cardiovascular risk factor in patients starting dialysis therapy. PMID- 12631363 TI - Proteinuria and the risk of developing end-stage renal disease. AB - BACKGROUND: Dipstick urinalysis for proteinuria and hematuria has been used to screen renal disease, but evidence of the clinical impact of this test on development of end-stage renal disease (ESRD) is lacking. METHODS: We assessed development of ESRD through 2000 in 106,177 screened patients (50,584 men and 55,593 women), 20 to 98 years old, in Okinawa, Japan, who participated in community-based mass screening between April 1983 and March 1984. We used data from the Okinawa Dialysis Study Registry to identify ESRD patients. Multivariate logistic analyses were performed to calculate adjusted odds ratio and 95% confidence interval (95% CI) for the significance of proteinuria and hematuria on the risk of developing ESRD with confounding variables such as age, gender, blood pressure, and body mass index. A similar analysis was repeated in a subgroup of screened patients in whom serum creatinine data existed. RESULTS: During 17 years of follow-up, 420 screened persons (246 men and 174 women) entered the ESRD program. We identified a strong, graded relationship between ESRD and dipstick urinalysis positive for proteinuria; adjusted odds ratio (95% CI) was 2.71 (2.51 to 2.92, P < 0.001). Similar trends were observed after adding serum creatinine data. Compared with dipstick-negative proteinuria, adjusted odds ratio (95% CI) of proteinuria (1+) was 1.93 (1.53 to 2.41, P < 0.001) in men and 2.42 (1.91 to 3.06, P < 0.001) in women. CONCLUSION: Proteinuria was a strong, independent predictor of ESRD in a mass screening setting. Even a slight increase in proteinuria was an independent risk factor for ESRD. Therefore, asymptomatic proteinuria warrants further work-up and intervention. PMID- 12631364 TI - The Fc gamma RIIIA-F158 allele is a risk factor for the development of lupus nephritis: a meta-analysis. AB - BACKGROUND: The Fc gamma RIIIA-V/F158 polymorphism affects immunoglobulins (Ig)G1 and IgG3-binding capacity and may modulate the expression of renal disease in patients with systemic lupus erythematosus (SLE). We aimed to determine whether this polymorphism confers risk for the development of lupus nephritis and SLE in general. METHODS: A meta-analysis was performed based on the Medline and Embase databases (last update, August 2002), perusal of abstracts from major meetings (1999 to 2001), assessment of bibliographies of pertinent articles, and additional data gathered after contact with primary investigators. RESULTS: A total of 16 comparisons from 11 studies involving V/F158 genotyping of 1154 patients with lupus nephritis, 1261 SLE patients without nephritis, and 1455 disease-free controls were included. Comparison of lupus nephritis patients with non-nephritis SLE subjects revealed a significant overrepresentation of the low binding F158 allele among patients who developed renal disease [odds ratio (OR) 1.20, 95% confidence interval (95% CI) 1.06 to 1.36, P = 0.003)], without significant between-study heterogeneity. FF homozygotes had the highest risk of renal disease as compared to VV homozygotes (OR 1.47, 95% CI 1.11 to 1.93, P = 0.006). It was uncertain whether the F158 allele influenced susceptibility to SLE per se (OR 1.19, 95% CI 0.99 to 1.43, P = 0.063 for SLE patients without nephritis versus disease-free controls; 0.01 < P < 0.10 for heterogeneity) and the observed trend for an association was driven mostly by the smaller studies (P = 0.058 for publication bias). No such bias was detected for analyses on susceptibility to lupus nephritis. CONCLUSION: The Fc gamma RIIIA-V/F158 polymorphism has a significant impact on the development of lupus nephritis. PMID- 12631365 TI - Paricalcitol versus calcitriol in the treatment of secondary hyperparathyroidism. AB - BACKGROUND: Management of secondary hyperparathyroidism has included the use of active vitamin D or vitamin D analogs for the suppression of parathyroid hormone (PTH) secretion. Although, these agents are effective, therapy is frequently limited by hypercalcemia, hyperphosphatemia, and/or elevations in the calcium phosphorus (Ca x P) product. In clinical studies, paricalcitol was shown to be effective at reducing PTH concentrations without causing significant hypercalcemia or hyperphosphatemia as compared to placebo. A comparative study was undertaken in order to determine whether paricalcitol provides a therapeutic advantage to calcitriol. METHODS: A double-blind, randomized, multicenter study comparing the safety and effectiveness of intravenous paricalcitol and calcitriol in suppressing PTH concentrations in hemodialysis patients was performed. A total of 263 randomized patients were enrolled at domestic and international sites. Following the baseline period, patients with serum Ca x P < 75, and a PTH level > or =300 pg/mL were randomly assigned to receive either paricalcitol or calcitriol in a dose-escalating fashion for up to 32 weeks. Dose adjustments were based on laboratory results for PTH, calcium, and Ca x P. The primary end point was the greater than 50% reduction in baseline PTH. Secondary end points were the occurrence of hypercalcemia and elevated Ca x P product. RESULTS: Paricalcitol treated patients achieved a > or =50% reduction from baseline PTH significantly faster than did the calcitriol-treated patients (P = 0.025) and achieved a mean reduction of PTH into a desired therapeutic range (100 to 300 pg/mL) at approximately week 18, whereas the calcitriol-treated patients, as a group, were unable to achieve this range. Moreover, paricalcitol-treated patients had significantly fewer sustained episodes of hypercalcemia and/or increased Ca x P product than calcitriol patients (P = 0.008). CONCLUSION: Paricalcitol treatment reduced PTH concentrations more rapidly with fewer sustained episodes of hypercalcemia and increased Ca x P product than calcitriol therapy. PMID- 12631366 TI - Nutrition in hemodialysis patients previously on a supplemented very low protein diet. AB - BACKGROUND: Nutritional safety of protein-restricted diets in patients with chronic renal failure is controversial. In the present study, we have assessed the evolution of nutritional status after initiation of hemodialysis in patients previously treated by a supplemented very low protein diet (SVLPD). METHODS: Nutritional data were prospectively collected during the first year of hemodialysis from 15 consecutive patients treated with a SVLPD (0.3 g protein/kg/day supplemented with essential amino acids, calcium, iron, and vitamins) and compared to 15 age- and gender-matched end-stage renal disease (ESRD) patients previously on a less-restricted diet (0.90 +/- 0.21 g protein/kg/day) who started hemodialysis during the same period. Dual-energy x ray absorptiometry (DEXA) was used to assess body composition at 0, 6, and 12 months. Hemodialysis prescriptions, biologic data and 3-day food records were collected every 3 months. RESULTS: Protein intake was higher than 1.2 g/kg/day in both groups as soon as 3 months after the start of hemodialysis. Albumin and prealbumin increased significantly during the first 6 months in all patients. Body mass index (BMI) increased in all patients (+0.97 +/- 1.31 kg/m2; P < 0.001) reflecting a gain in fat mass in the overall population (+2.36 +/- 2.94 kg/m2; P < 0.001) while lean body mass remained stable overall. CONCLUSION: Once on hemodialysis, SVLPD patients rapidly increased protein intake. Nutritional status improved in all patients, with a gain in fat mass in all, and a gain in lean body mass in SVLPD men only. These data indicate that treatment with a SVLPD prior to hemodialysis initiation is nutritionally safe. PMID- 12631367 TI - The risk of developing end-stage renal disease in patients with type 2 diabetes and nephropathy: the RENAAL study. AB - BACKGROUND: Diabetic nephropathy has become the single most important cause of end-stage renal disease (ESRD) worldwide. Strategies to slow the rate of loss of renal function in these patients have been developed. We examined the risk factors that predict loss of kidney function (doubling of serum creatinine) or ESRD (dialysis or transplantation) in patients with type 2 diabetes in whom blood pressure was controlled. METHODS: We evaluated risk factors for doubling of serum creatinine or the development of ESRD in the Reduction of End Points in NIDDM with the Angiotensin II Receptor Antagonist Losartan (RENAAL) study, which included 1513 patients with type 2 diabetes and nephropathy. RESULTS: Univariate analyses demonstrated a group of 23 risk factors that significantly predicted doubling of serum creatinine or ESRD. From these univariate analyses, a multivariate model was developed that demonstrated four independent risk factors: proteinuria, serum creatinine, serum albumin, and hemoglobin level. Proteinuria was the strongest and most consistent risk factor. The multivariate risk model was derived from only the placebo group and was similar to that derived for the total population, suggesting that the risk predictors for progression of kidney disease were independent of therapy. CONCLUSION: After control of blood pressure in type 2 diabetic patients with nephropathy, proteinuria, degree of renal failure, serum albumin, and hemoglobin level are independent risk factors that predict renal outcomes. The level of proteinuria proved to be the most important risk for progressive kidney injury in these diabetic patients. PMID- 12631369 TI - Nonheart-beating kidney donation: current practice and future developments. AB - BACKGROUND: Nonheart-beating kidney donation (NHBD) is gaining acceptance as a method of donor pool expansion. However, a number of practitioners have concerns over rates of delayed graft function, acute rejection, and long-term graft survival. The ethical issues associated with NHBD are complex and may be a further disincentive. Tailored strategies for preservation, viability prediction, and immunosuppression for kidneys from this source have the potential to maximize the number of available organs. This review article presents the current practice of NHBD kidney transplantation, examines the results and draws comparisons with cadaveric kidneys, and explores some areas of potential development. METHODS: A review of the current literature on NHBD kidney donation was performed. RESULTS: The renewed interest in NHBD kidneys is driven by a continuing shortfall in available organs. Those centers involved in NHBD report an increase in kidney transplants of the order of 16% to 40% and there is no evidence that the financial costs are higher with NHBDs. The majority of experience comes from Maastricht category 2 NHBDs, where an estimation of warm time is possible. This is generally limited to 40 minutes. There are variations in the technique for kidney preservation prior to retrieval, but most centers use an aortic balloon catheter. Much work has looked at the ideal technique for kidney preservation prior to implantation. Evidence suggests that machine perfusion produces the best initial function rates, decreased use of adjuvant immunotherapy and fewer haemodialysis sessions than static cold storage. CONCLUSION: Despite being associated with poorer initial graft function, the long-term allograft survival of NHBD kidneys does not differ from the results of transplantation from cadaveric kidneys. Further, serum creatinine levels are generally equivalent. Constant reassessment of the ethical issues is required for donation to be increased while respecting public concerns. Use of viability assessment and tailoring of immune suppression for NHBD kidneys may allow a further increase in donation from this source. PMID- 12631368 TI - Risk factors for renal glomerular and vascular changes in an autopsy-based population survey: the Hisayama study. AB - BACKGROUND: Information of the effect of cardiovascular risk factors on renal glomerular and vascular changes is scarce in the general population. METHOD: Between 1962 and 1994, 1394 autopsies were performed in Hisayama, for a total autopsy rate of 80%. Of these, 839 individuals who preserved adequate renal tissues and had recent health examinations data before death were eligible for the present study. We examined the degree of glomerular sclerosis, renal arteriolar hyalinosis, and arteriosclerosis, and evaluated their risk factors by means of a logistic regression model. RESULTS: The development of glomerular sclerosis, arteriolar hyalinosis, and arteriosclerosis were 16%, 16%, and 18% in men, respectively, and 27%, 15%, and 24% in women, respectively. All these frequencies increased linearly with advancing age. In the multivariate analysis, both age and systolic blood pressure were significant independent risk factors for almost all these glomerular and vascular changes. In addition, glucose intolerance and proteinuria for men were found to be significant risk factors for glomerular sclerosis. Elevated total cholesterol levels significantly increased the risk of arteriolar hyalinosis in men. Electrocardiogram (ECG) abnormalities were an independent risk factor for arteriosclerosis in both men and women, and proteinuria was an additional risk factor in women. Alcohol intake tended to have a protective effect on glomerular sclerosis and arteriosclerosis in women. CONCLUSION: Our data confirmed that age and systolic blood pressure are common risk factors for all glomerular and renal vascular changes in the general population. In addition, glucose intolerance, total cholesterol, ECG abnormalities, and proteinuria affect either glomerular or vascular changes. PMID- 12631370 TI - Myofibroblastic conversion of mesothelial cells. AB - BACKGROUND: The continuous chemical, physical, and inflammatory insults of prolonged continuous ambulatory peritoneal dialysis (CAPD) incite mesothelial cell responses, which may result in peritoneal fibrosis. The transforming growth factor-beta (TGF-beta), especially the isoform TGF-beta 1, has long been known to play crucial role in the fibrogenic process. Although several studies have implicated TGF-beta in peritoneal fibrosis, the underlying mechanism has not been completely elucidated. METHODS: To test the effects of exogenous TGF-beta 1 on mesothelial cells, we assessed cytoarchitectural changes of human peritoneal mesothelial cells (HPMC) in in vitro culture by light, immunofluorescent, electron and immunoelectron microscopy, and differential gene expression analysis using semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) and cDNA expression array assays. RESULTS: The TGF-beta 1-induced myofibroblastic conversion was a transdifferentiation process resulting in characteristic myofibroblastic phenotype that included prominent rough endoplasmic reticuli (rER) with dilated cisternas, conspicuous smooth muscle actin (SMA) myofilaments, frequent intercellular intermediate and gap junctions, and active deposition of extracellular matrix (ECM) and formation of fibronexus. The gene expression array analysis revealed complex modulation of gene expression involving cytoskeletal organization, cell adhesion, ECM production, cell proliferation, innate immunity, cytokine/growth factor signaling, cytoprotection, stress response, and many other essential metabolic processes in mesothelial cells. CONCLUSION: This report describes myofibroblastic conversion of mesothelial cells, a previously undefined, yet frequently speculated, cell adaptive or pathogenic process. Our study helps to elucidate the complex molecular and cellular events involved in myofibroblastic conversion of mesothelial cells. We propose that differentiated epithelial cells of mesothelium convert or transdifferentiate into myofibroblasts, which implies the recruitment of fibrogenic cells from mesothelium during serosal inflammation and wound healing. PMID- 12631371 TI - Studies on dialysate mixing in the Genius single-pass batch system for hemodialysis therapy. AB - BACKGROUND: The Genius single-pass batch system for hemodialysis contains a closed reservoir and dialysate circuit of 75 L dialysate. The unused dialysate is withdrawn at the top of the reservoir and the spent fluid is reintroduced into the container at the bottom. Although it has been claimed that both fractions remain unmixed during the dialysis session, no direct proof of this assumption has yet been provided. In the present study, we investigated whether contamination of the unused dialysate with uremic solutes occurred and at which time point it began. Two different dialysate temperatures were compared. METHODS: Ten chronic hemodialysis patients were dialyzed twice with the Genius system, with dialysate prepared at 37 degrees C and 38.5 degrees C, respectively. The sessions lasted 270 minutes with blood/dialysate flow set at 300 mL/min. Dialysate was sampled at 5, 60, 180, 210, 225, 230, 235, 240, 255, and 270 minutes both from the inlet and outlet dialysate line and blood was sampled from the arterial line predialysis, after 4 hours, and postdialysis. All samples were tested for osmolality, urea, creatinine, p-cresol, hippuric acid, and indoxyl sulfate. RESULTS: Uremic solutes appeared in the inlet dialysate line between 3 hours 50 minutes and 4 hours 10 minutes after the start of dialysis, corresponding to 68.6 and 74.7 L spent dialysate, respectively (37 degrees C vs. 38.5 degrees C; P = NS). No difference in the amount of removed solutes and in the serum levels was observed between 37 degrees C and 38.5 degrees C. A Kt/V of 1.17 +/- 0.20 and 1.18 +/- 0.26, respectively, was reached with the 37 degrees C and 38.5 degrees C dialysate temperature (P = NS). CONCLUSION: Contamination with uremic solutes occurred at the dialysate inlet only near the end of the session when small quantities of fresh dialysate were left in the container. Differences in dialysate temperature did not result in a different separation between used and unused dialysate, or in differences in removal of toxins or Kt/V. PMID- 12631372 TI - Depolymerized holothurian glycosaminoglycan (DHG), a novel alternative anticoagulant for hemodialysis, is safe and effective in a dog renal failure model. AB - BACKGROUND: Depolymerized holothurian glycosaminoglycan (DHG) is a new agent with anticoagulant properties quite different from those of unfractionated heparin (UFH) and low-molecular-weight heparin (LMWH) in terms of antithrombin III dependency, and exerts an antithrombotic effect with less bleeding than UFH and LMWH in vivo. In this study, the anticoagulant and hemorrhagic effects of DHG were investigated on hemodialysis in a dog model of renal failure and compared with those of UFH, LMWH, and nafamostat mesilate (FUT). METHODS: The dog renal failure model was prepared by 7/8 renal artery ligation. Effectiveness was based on completion of 3-hour hemodialysis, no marked clot deposition in the extracorporeal circuit, and permeability of blood urea nitrogen (BUN) and creatinine. Template bleeding was measured by determining the hemoglobin content of the blood from the wound. RESULTS: DHG induced no major bleeding or clot formation during 3-hour hemodialysis, in contrast to UFH and LMWH, each of which induced marked bleeding. These glycosaminoglycans (GAGs) were equally effective in decreasing plasma levels of BUN and creatinine. On the other hand, dogs treated with FUT failed to complete 3-hour hemodialysis. These anticoagulants prolonged activated partial thromboplastin time (APTT) to different extents and GAGs prolonged thrombin clotting time markedly but FUT did not. CONCLUSION: Our findings suggest that thrombin clotting time prolongation can contribute to prevention of clot formation in extracorporeal circuits, and the non-antithrombin III-dependent activities of DHG may be related to its low risk of hemorrhage for hemodialysis. DHG appears to be promising as an alternative anticoagulant with low risk of hemorrhage for hemodialysis. PMID- 12631373 TI - Effect of icodextrin on volume status, blood pressure and echocardiographic parameters: a randomized study. AB - Overhydration is a risk factor for hypertension and left ventricular hypertrophy in peritoneal dialysis patients. Recently, a high prevalence of subclinical overhydration was observed in peritoneal dialysis patients. Aim of the present open-label randomized study was to assess the effect of a icodextrin 7.5% solution on fluid status [extracellular water (ECW) bromide dilution], blood pressure regulation (24-hour ambulatory measurements) and echocardiographic parameters during a study period of 4 months, and to relate the effect to peritoneal membrane characteristics (dialysate/plasma creatinine ratio). Forty peritoneal dialysis patients (22 treated with icodextrin, 18 controls) were randomized to either treatment with icodextrin during the long dwell or standard glucose solutions. Thirty-two patients (19 treated with icodextrin, 13 controls] completed the study. The use of icodextrin resulted in a significant increase in daily ultrafiltration volume (744 +/- 767 mL vs. 1670 +/- 1038 mL; P = 0.012) and a decrease in ECW (17.5 +/- 5.2 L vs. 15.8 +/- 3.8 L; P = 0.035). Also the change in ECW between controls and patients treated with icodextrin was significant ( 1.7 +/- 3.3 L vs. +0.9 +/- 2.2 L; P = 0.013). The effect of icodextrin on ECW was not related to peritoneal membrane characteristics, but significantly related to the fluid state of the patients (ECW:height) (r = -0.72; P < 0.0001). Left ventricular mass (LVM) decreased significantly in the icodextrin-treated group (241 +/- 53 grams vs. 228 +/- 42 grams; P = 0.03), but not in the control group. In this randomized open-label study, the use of icodextrin resulted in a significant reduction in ECW and LVM. The effect of icodextrin on ECW was not related to peritoneal membrane characteristics, but was related to the initial fluid state of the patient. PMID- 12631374 TI - Heme oxygenase-1 attenuates ischemia/reperfusion-induced apoptosis and improves survival in rat renal allografts. AB - BACKGROUND: Kidneys can be preserved only for a limited time without jeopardizing graft function and survival. Induction of heat shock proteins (HSPs) can protect against ischemia/reperfusion (I/R) injury. Therefore, we investigated whether the induction of the HSP, heme oxygenase-1 (HO-1), improves outcome following isotransplantation after an extended period of cold storage. METHODS: Rats were subjected to heat preconditioning (HP; 42 degrees C for 20 minutes). Kidneys harvested after 24 hours, were preserved in cold University of Wisconsin (UW) solution at 4 degrees C for 45 hours and transplanted into bilateral nephrectomized rats. Cobalt protoporphyrin (CoPP) was administered in another group of animals in order to induce HO-1 pharmacologically, while other groups of animals received the HO-1 inhibitor, tin protophorphyrine (SnPP), following HP or CoPP. RESULTS: Cold ischemia caused a complete attenuation of graft function within 3 days following transplantation and subsequent death of all animals, whereas HP protected graft function and five of nine rats survived for 3 weeks. HP inhibited the induction of osteopontin and induced the expression of HO-1, HSP 70 and 90, and the antiapoptotic factor Bcl-XL. Grafts exposed to HP were protected against structural I/R injuries as revealed by histologic assessment using a semiquantitative score. Furthermore, induction of apoptosis was attenuated and activation of caspase-3 was inhibited. Comparable results were observed after administration of CoPP, whereas SnPP inhibited the effects of HP and CoPP. CONCLUSION: HP or administration of CoPP induced both HO-1, preserved kidney graft function, and prevented postreperfusion apoptosis after cold preservation. PMID- 12631375 TI - Prevention of biofilm formation in dialysis water treatment systems. AB - BACKGROUND: Biofilm formations in dialysis systems may be relevant because they continuously release bacterial compounds and are resistant against disinfection. The aim of the study was to compare the development of biofilm between a water treatment system based on a single reverse osmosis unit producing purified dialysate water [bacterial count, 350 colony-forming unit (CFU)/L] (center A) and a water treatment system based on double reverse osmosis and electric deionization, which is continuously disinfected with ultraviolet light and treated with ozone once a week (bacterial count, 1 CFU/L) (center B). METHODS: During a period of 12 weeks, biofilm formation was studied in the tubing segment between the water piping and the dialysis module, using four dialysis monitors in each center. On a weekly basis, tubing samples of 5 cm length (N = 96) were taken under aseptic conditions and investigated for microbiologic contamination [cystine lactose electrolyte-deficient (CLED) Agar], endotoxin levels [limulus amoeben lysate (LAL) gel test, cutoff value, 0.0125 EU/mL], and biofilm formation [electron scanning microscopy (SEM)]. RESULTS: In center A, tube cultures were positive (>100 CFU/mL) in 16% of samples at 22 degrees C and 37 degrees C, compared to 3% of samples of center B (P < 0.05; chi-square). Endotoxin levels were positive in 76% of the tubing samples of center A and negative in all of the samples of center B (P < 0.05). Biofilm was present in 91.7% of the samples of center A (Fig. 1), and only present in one sample (taken after 9 weeks) of center B (P < 0.05) (Fig. 2). In center A, biofilm formation was already observed after 1 week. CONCLUSION: In contrast to a standard water treatment system producing purified water, the use of a system producing highly purified water, which is also treated with regular disinfection procedures, leads to a significant reduction in biofilm formation, bacterial growth, and endotoxin levels in a highly vulnerable part of a water treatment system. PMID- 12631376 TI - Dialysis delayed is death prevented: a clinical perspective on the RENAAL study. PMID- 12631377 TI - Smoking and dialysis: a dreadful scenario for the cardiovascular system? PMID- 12631378 TI - A role for macrophages in mediating tubular cell apoptosis? PMID- 12631379 TI - Interactions between growth factors in the kidney: implications for progressive renal injury. PMID- 12631380 TI - Update on vitamin D and its newer analogues. PMID- 12631381 TI - Adequacy of dialysis revisited. PMID- 12631383 TI - Beta-catenin: a transforming actor on many stages. AB - Mutations and deletions that result in the stabilization of beta-catenin are frequently found in a number of tumors, including those of the colon, the liver and the ovary, but are less frequently found in breast cancer. To investigate and understand the molecular nature of cell-specific beta-catenin signaling, experimental mouse genetics has been employed extensively. Gain-of-function and loss-of-function mutations have provided evidence that beta-catenin plays essential roles in development and tumorigenesis. Specifically, the Wnt/beta catenin signaling pathway controls cell fate decisions throughout development, and a unique role in differentiated epithelia has emerged. Not only beta-catenin, but also the activation of other components of this pathway in differentiated mammary epithelium and prostate epithelium of transgenic mice can induce neoplasias and transdifferentiation to squamous metaplasias. This suggests that the Wnt/beta-catenin pathway is dominant over existing differentiation programs and can impose an epidermal fate or neoplasias onto a variety of cell types. Although there is evidence for a contextual specificity of the Wnt signaling, the experimental systems and designs used in different studies probably influence the cellular responses. PMID- 12631384 TI - Is there a role for Notch signalling in human breast cancer? AB - Aberrant Notch signalling has been observed in several human cancers, including acute T-cell lymphoblastic leukaemia and cervical cancer, and is strongly implicated in tumourigenesis. Unregulated Notch signalling in the mouse mammary gland leads to tumour formation. These results raise the possibility that Notch signalling might play a role in human breast cancer. There are currently few reports that address this question directly and this appears to be an area worthy of further investigation. PMID- 12631386 TI - Host microenvironment in breast cancer development: inflammatory and immune cells in tumour angiogenesis and arteriogenesis. AB - Breast cancer progression is associated with and dependent upon robust neovascularization. It is becoming clear that tumour-associated 'normal' cells, such as immune/inflammatory cells, endothelial cells and stromal cells, conspire with cancer cells in promoting this process. In particular, infiltrating immune/inflammatory cells secrete a diverse repertoire of growth factors and proteases that enable them to enhance tumour growth by stimulating angiogenesis and, as we suggest here, by promoting 'tumour arteriogenesis' - enlargement of feeding vessels supplying the expanding tumour capillary bed. Macrophages and their chemoattractants (e.g. macrophage chemoattractant protein-1) are critical for the arteriogenic process in ischaemia, and probably also in breast neoplasia. A better understanding of these various cellular and molecular constituents of breast cancer neovascularization may be useful in designing more effective therapies. PMID- 12631385 TI - The prolyl isomerase Pin1 in breast development and cancer. AB - The prolyl isomerase Pin1 specifically isomerizes certain phosphorylated Ser/Thr Pro bonds and thereby regulates various cellular processes. Pin1 is a target of several oncogenic pathways and is overexpressed in human breast cancer. Its overexpression can lead to upregulation of cyclin D1 and transformation of breast epithelial cells in collaboration with the oncogenic pathways. In contrast, inhibition of Pin1 can suppress the transformation of breast epithelial cells. In addition, Pin1 knockout in mice prevents massive proliferation of breast epithelial cells during pregnancy. Pin1 plays a pivotal role in breast development and may be a promising new anticancer target. PMID- 12631387 TI - Breast cancer cell lines: friend or foe? AB - The majority of breast cancer research is conducted using established breast cancer cell lines as in vitro models. An alternative is to use cultures established from primary breast tumours. Here, we discuss the pros and cons of using both of these models in translational breast cancer research. PMID- 12631388 TI - Trastuzumab, an appropriate first-line single-agent therapy for HER2 overexpressing metastatic breast cancer. AB - Overexpression of the HER2/Neu (ErbB2) proto-oncogene is associated with breast cancer progression and poor patient prognosis. Herceptin (trastuzumab) is a humanized IgG1 against the ectodomain of the HER2 receptor. In combination with chemotherapy, it induces regression of HER2-overexpressing metastatic breast tumors and prolongs patient survival. Single-agent Herceptin in patients with HER2-amplified breast tumors also induces a definite objective response and clinical benefit rates, and is well tolerated. These data suggest that Herceptin is an effective first-line single-agent therapy for a predictable cohort of metastatic breast cancers and can therefore be used as a platform for therapeutic discovery against tumors that overexpress HER2. PMID- 12631390 TI - New insights into the biological function of BRCA2 from its structural interactions. PMID- 12631391 TI - 25th Annual San Antonio Breast Cancer Symposium, San Antonio, Texas, USA, 10-14 December 2002 Update on preclinical and translational research. AB - The Annual San Antonio Breast Cancer Symposium is one of the largest regular conferences devoted to breast cancer research and treatment. In particular, it provides a forum in which to discuss the more translational aspects of current basic research, and the 2002 meeting was no exception. Growth factor pathways and endocrine resistance, cancer genomics and the clinical applications of proteomics were three of the major topics for discussion. Presentations on genetic susceptibility and the development of prognostic and predictive markers also created much interest. PMID- 12631392 TI - 25th Annual San Antonio Breast Cancer Symposium, San Antonio, Texas, USA, 10-14 December 2002 Update on clinical research. AB - The Annual San Antonio Breast Cancer Symposium has become a key forum for the presentation and discussion of both translational scientific aspects as well as clinical aspects of breast cancer care. In this report of the 25th Annual Meeting, an update of the salient clinical data is presented. The findings of the CALGB 9741 trial, an updated analysis of the Arimidex, Tamoxifen, Alone or in Combination study, and other significant paper and poster presentations are discussed. Summaries are also given of the clinical plenary lectures and minisymposia on adjuvant therapy and aromatase inhibitors. PMID- 12631389 TI - Host microenvironment in breast cancer development: epithelial-mesenchymal transition in breast cancer development. AB - The epithelial-mesenchymal transition (EMT) is a developmental mechanism of crucial importance in establishing the body plan in many multicellular organisms. Several transduction pathways controlling the various steps of the morphological transition have been identified by molecular analyses of this process in cell lines and in vivo. The newly formed mesenchymal cells can exhibit locomotory and invasive phenotypes, suggesting that EMTs contribute to the progression of carcinoma. Diverse evidence indicates that EMT subprograms are involved in the appearance of different breast carcinoma types. Several normal and malignant breast cell lines are currently being analyzed to define key steps in EMT and to identify candidate genes. DNA profiling technology is also being applied to uncover pathways that lead to a metastatic phenotype. PMID- 12631393 TI - Polarity determination in breast tissue: desmosomal adhesion, myoepithelial cells, and laminin 1. AB - In all epithelial organs, apicobasal polarity determines functional integrity and contributes to the maintenance of tissue and organ specificity. In the breast, the functional unit is a polar double-layered tube consisting of luminal epithelial cells surrounded by myoepithelial cells and a basement membrane. It is far from clear how this double-layered structure is established and how polarity is maintained. Two recent papers have shed some light onto this intriguing problem in mammary gland biology. The results point to desmosomes and laminin 1 as having crucial roles. However, some questions remain. PMID- 12631394 TI - Waist-hip ratio and breast cancer risk in urbanized Nigerian women. AB - BACKGROUND: The aim of this study was to examine the relationship between waist hip ratio and the risk of breast cancer in an urban Nigerian population. METHODS: Between March 1998 and August 2000, we conducted a case-control study of hospital based breast cancer patients (n = 234) and population-based controls (n = 273) using nurse interviewers in urban Southwestern Nigeria. RESULTS: Multivariable logistic regression showed a significant association between the highest tertile of waist-hip ratio and the risk of breast cancer (odds ratio= 2.67, 95% confidence interval = 1.05-6.80) among postmenopausal women. No association was found in premenopausal women. CONCLUSION: The present study, the first in an indigenous African population, supports other studies that have shown a positive association between obesity and breast cancer risk among postmenopausal women. PMID- 12631395 TI - Id-1 is not expressed in the luminal epithelial cells of mammary glands. AB - BACKGROUND: The family of inhibitor of differentiation/DNA binding (Id) proteins is known to regulate development in several tissues. One member of this gene family, Id-1, has been implicated in mammary development and carcinogenesis. Mammary glands contain various cell types, among which the luminal epithelial cells are primarily targeted for proliferation, differentiation and carcinogenesis. Therefore, to assess the precise significance of Id-1 in mammary biology and carcinogenesis, we examined its cellular localization in vivo using immunohistochemistry. METHODS: Extracts of whole mammary glands from wild type and Id-1 null mutant mice, and tissue sections from paraffin-embedded mouse mammary glands from various developmental stages and normal human breast were subjected to immunoblot and immunohistochemical analyses, respectively. In both these procedures, an anti-Id-1 rabbit polyclonal antibody was used for detection of Id-1. RESULTS: In immunoblot analyses, using whole mammary gland extracts, Id 1 was detected. In immunohistochemical analyses, however, Id-1 was not detected in the luminal epithelial cells of mammary glands during any stage of development, but it was detected in vascular endothelial cells. CONCLUSION: Id-1 is not expressed in the luminal epithelial cells of mammary glands. PMID- 12631396 TI - C-myc, not HER-2/neu, can predict recurrence and mortality of patients with node negative breast cancer. AB - BACKGROUND: At present, node-negative, high-risk breast cancer patients cannot be identified with sufficient accuracy. Consequently, further strong prognostic factors are needed. METHODS: Among 181 node-negative breast cancer (NNBC) patients, c-myc and HER-2/neu oncogenes were identified prospectively using double differential PCR. The possible impact of amplification of those oncogenes on disease-free survival (DFS) and overall survival was examined. Furthermore, the possible effects of adjuvant therapies on rate of recurrence and mortality in oncogene-amplified NNBC patients were investigated. RESULTS: The prevalence rates for amplification of c-myc and HER-2/neu were 21.5% and 30.4%, respectively. On univariate analysis, c-myc-amplified NNBCs were associated with significantly shorter DFS at 36 months after the initial diagnosis (85.3% versus 97.3%). As compared with nonamplified cancers, HER-2/neu-amplified NNBCs did not exhibit any significant differences after 36 months and 95 months. Multivariate analysis indicated that c-myc amplification and tumour size, in contrast to HER-2/neu amplification, oestrogen receptor status, grading and age, were the only independent parameters for DFS. During the period of observation, we found no evidence for an impact of amplification of the oncogenes on overall survival in all cases. With respect to various adjuvant systemic therapies such as chemotherapy (cyclophosphamide, methotrexate, 5-fluorouracil; fluorouracil, epirubicin, cyclophosphamide) and endocrine therapy (tamoxifen), no significant differences were identified in oncogene-amplified NNBC patients in terms of DFS and overall survival. However, those c-myc-amplified NNBC patients who did not receive adjuvant systemic therapy exhibited significantly shorter DFS and overall survival as compared with c-myc-nonamplified patients. CONCLUSION: C-myc amplification appears to be a strong prognostic marker with which to predict early recurrence in NNBC patients. C-myc-amplified NNBC patients without adjuvant systemic therapy experienced shorter DFS and overall survival. PMID- 12631398 TI - CYP17 genetic polymorphism, breast cancer, and breast cancer risk factors. AB - BACKGROUND: Findings from previous studies regarding the association between the CYP17 genotype and breast cancer are inconsistent. We investigated the role of the MspAI genetic polymorphism in the 5' region of CYP17 on risk of breast cancer and as a modifier of reproductive risk factors. METHODS: Questionnaire and genotyping data were obtained from a population-based, case-control study of premenopausal (n = 182) and postmenopausal (n = 214) European-American Caucasian women in western New York. Cases and controls were frequency matched by age and by county of residence. Odds ratios and 95% confidence intervals were used to estimate relative risks. RESULTS: The CYP17 genotype was not associated with breast cancer risk; however, controls with the A2/A2 genotype (associated with higher estrogens) had earlier menarche and earlier first full-term pregnancy. Premenopausal women with A1/A1 genotypes, but not with A2 alleles, were at significantly decreased risk with late age at menarche (odds ratio = 0.37, 95% confidence interval = 0.14-0.99), and at increased risk with late age at first full-term pregnancy (odds ratio = 4.30, 95% confidence interval = 1.46-12.67) and with use of oral contraceptives (odds ratio = 3.24, 95% confidence interval = 1.08-9.73). Associations were weaker among postmenopausal women. CONCLUSION: These results suggest that the effects of factors that may alter breast cancer risk through a hormonal mechanism may be less important among premenopausal women with putative higher lifetime exposures to circulating estrogens related to the CYP17 A2 allele. PMID- 12631397 TI - Akt kinases in breast cancer and the results of adjuvant therapy. AB - BACKGROUND: The serine/threonine kinase Akt, or protein kinase B, has recently been a focus of interest because of its activity to inhibit apoptosis. It mediates cell survival by acting as a transducer of signals from growth factor receptors that activate phosphatidylinositol 3-kinase. METHODS: We analysed the expression of the isoforms Akt1 and Akt2 as well as phosphorylated Akt (pAkt) by immunohistochemistry in frozen tumour samples from 280 postmenopausal patients who participated in a randomised trial comparing cyclophosphamide-methotrexate-5 fluorouracil chemotherapy and postoperative radiotherapy. The patients were simultaneously randomised to tamoxifen or to no endocrine treatment. RESULTS: Marked staining was found in 24% of the tumours for Akt1, but in only 4% for Akt2. A low frequency of Akt2-positive cells (1-10%) was observed in another 26% of the tumours. pAkt was significantly associated with both Akt1 and Akt2 expression. Overexpression of erbB2 correlated significantly with pAkt (P = 0.0028). The benefit from tamoxifen was analysed in oestrogen receptor (ER) positive patients. Patients with a negative status of Akt (no overexpression of Akt1, Akt2 or pAkt) showed significant benefit from tamoxifen. The relative rate of distant recurrence, with versus without tamoxifen, was 0.44 (95% confidence interval [CI], 0.25-0.79) for ER+/Akt1- patients, while it was 0.72 (95% CI, 0.34 1.53) for ER+/Akt1+ patients. The difference in rate ratio did not reach statistical significance. The rate of locoregional recurrence was significantly decreased with radiotherapy versus chemotherapy for Akt-negative patients (rate ratio, 0.23; 95% CI, 0.08-0.67; P = 0.0074), while no benefit was evident for the Akt-positive subgroup (rate ratio, 0.77; 95% CI, 0.31-1.9; P = 0.58). The interaction between Akt and the efficacy of radiotherapy was significant in multivariate analysis (P = 0.042). CONCLUSION: Activation of the Akt pathway is correlated with erbB2 overexpression in breast cancer. The results suggest that Akt may predict the local control benefit from radiotherapy. PMID- 12631399 TI - A candidate metastasis-associated DNA marker for ductal mammary carcinoma. AB - BACKGROUND: Molecular genetic markers to identify the 13% lymph node-negative mammary carcinomas that are prone to develop metastases would clearly be of considerable value in indicating those cases in need of early aggressive therapy. METHODS: Representational difference analysis was used in an attempt to identify genetic alterations related to breast cancer metastasis by comparing genomic DNA from microdissected normal cells and from metastatic cells of ductal breast carcinoma patients. RESULTS: Representational difference analysis products yielded 10 unique metastasis-associated DNA sequences (MADS), i.e. products apparently lost in metastatic cell DNA. Of these sequences, MADS-IX was found to be lost in the transition from primary to metastasis in two out of five ductal breast carcinoma cases. This sequence was localized on chromosome 10q21 by radiation hybrid mapping and fluorescence in situ hybridization. The PTEN gene, which is also located on chromosome 10q, was detected to be present by PCR in all five cases. On the contrary, a breast carcinoma cell line, HCC-1937, which has homozygous loss of a region encompassing the PTEN gene, showed the presence of MADS-IX. PCR screening of three additional breast carcinoma cell lines with known losses in specific chromosomal regions also showed the presence of MADS-IX. CONCLUSION: These data suggest that MADS-IX possibly is part of a novel candidate metastasis-associated gene located close to the PTEN gene on chromosome 10q. The first set of PCR screening in five patient samples indicates that it could be used as a molecular marker for ductal mammary metastasis. PMID- 12631401 TI - Keying in "key result areas". PMID- 12631400 TI - Adolescent diet and risk of breast cancer. AB - BACKGROUND: Early life exposures, including diet, have been implicated in the etiology of breast cancer. METHODS: A nested case-control study was conducted among participants in the Nurses' Health Study who completed a 24-item questionnaire about diet during high school. There were 843 eligible cases diagnosed between onset of study (1976) and before the return of the high school diet questionnaire (1986), who were matched 10:1 with controls on the basis of age. RESULTS: Women who had, during adolescence, a higher consumption of eggs, vegetable fat and fiber had a lower risk of breast cancer, whereas risk of breast cancer was increased among women who consumed more butter. CONCLUSIONS: A possible association of elements of adolescent diet with risk of breast cancer is reported, but the findings require confirmation in prospective study. PMID- 12631402 TI - Bone up on bone health. PMID- 12631403 TI - Protein and bone health: literature review and counselling implications. AB - For decades, public health promotion campaigns on bone health have emphasized the importance of adequate calcium and vitamin D intakes, as well as weight-bearing physical activity. However, no obvious consensus has emerged on the role of dietary protein. To identify what agreement does exist in the literature, in this article we review the theoretical basis for protein's role in bone health, assess some recent cross-sectional and prospective studies, and generate recommendations for practice. There is general agreement in the literature that higher protein intake increases urinary calcium loss; the body compensates for this loss by increasing calcium absorption in the gut, providing that calcium intake is sufficient. A possible explanation for calcium loss, the "acid-ash" hypothesis, is discussed, and suggestions are made about food choices that may counter the calciuric effect of protein. A survey of cross-sectional and prospective studies shows equivocal results, with confounding variables complicating the analysis. Both deficient and excessive protein intakes have been shown to affect bone health negatively, although lower and upper thresholds have not been determined. Practical advice on achieving bone health is given, with an emphasis on the use of Canada's Food Guide to Healthy Eating in setting dietary goals. PMID- 12631404 TI - Client-centred nutrition counselling: do we know what this means? AB - In Canada, professional standards mandate that dietitians should use a client centred approach to provide nutrition counselling services. Although most dietitians would probably agree that this is an important standard, how this mandate is translated into our daily practice is not always clear. The purpose of this paper is to explore the origins of the "client-centred approach" used in dietetic counselling. A historical review of selected dietetic literature is used to demonstrate the evolution of this term, the multiple meanings associated with it, the remaining ambiguity in dietetic practice today, and the need for further research. PMID- 12631405 TI - Dietary intake of older adults in the Kingston area. AB - The objectives of this research were to describe the dietary intake and identify risk factors for poor dietary intake in community-dwelling older adults living in the Kingston, Frontenac, and Lennox & Addington Health Unit area. Dietary intake information was collected from a convenience sample of 105 relatively healthy, active older adults (84 women, 21 men) using 24-hour recalls from three non consecutive days. Risk factors for poor dietary intake were identified through a structured interview. Multiple linear regression was used to generate a model to predict dietary intake, which was measured using a diet score based on Canada's Food Guide to Healthy Eating. Group averages reflected reasonable diet quality, but some subjects had very low nutrient intakes, particularly of zinc and vitamins B6, B12, and C. On average, women had a lower-than-recommended intake from all food groups, while men consumed adequate amounts of all food groups except milk products. Higher scores indicated better overall diet quality, and the following were significant predictors of a high diet score: "almost always" preparing one's own meals, food "almost always" or "sometimes/never" tasting good, eating lunch every day, and taking fewer prescription medications. This model requires validation with a larger and more diverse population of community dwelling older adults. PMID- 12631406 TI - Determining how low-income food shoppers perceive, understand, and use food labels. AB - A facilitated group discussion approach was used to explore how a group of eight low-income consumers perceive, understand, and use the current food label for food-shopping decisions. Meeting for one hour on three consecutive weeks in a community facility, the group discussed the total food package, the ingredient list alone and in combination with the nutrition-information panel, and nutrition claims. All sessions were recorded and later transcribed verbatim. Through inductive analysis, three broad categories (brand name, package feature, and nutrition-related information) and numerous underlying themes emerged from the data; a theoretical framework was proposed to explain the data and provide future direction. A number of barriers to effective use of the food label were identified, and some actionable tips for dietetic practice were suggested. PMID- 12631407 TI - The effect of food components on the absorption of P-gp substrates: a review. AB - P-glycoprotein (P-gp), a well characterized efflux mechanism which is functionally expressed in the intestinal epithelium, constitutes, along with intestinal metabolism, an important part of the biochemical barrier function of the intestinal mucosa. This efflux carrier may be responsible for limiting the bioavailability of several drugs after oral intake. Recently, increasing attention is being paid to the interaction of dietary components with the intestinal absorption of drugs. This review focuses on the modulating capacity of food components on the intestinal absorption of P-gp substrates. The possible P gp inhibitory effects of several dietary constituents are discussed. In addition, this review will also focus on the effect of several bioflavonoids on the P-gp mediated efflux of drugs. As the role of P-gp (and other efflux carriers, including multidrug resistance-associated proteins and breast cancer resistance protein) in limiting the bioavailability of drugs becomes more clear, more research is required firstly to identify the effect of dietary compounds on these efflux carriers and secondly to reveal the clinical relevance of this interaction. PMID- 12631408 TI - In-vitro evaluation of bioadhesion in particulate systems and possible improvement using interactive mixtures. AB - The bioadhesion of tablet components was tested using the fracture method (maximum tensile stress in detaching the sample from mucus membrane) and comparing traditional tablet specimens with powder monolayers. Both nonadhesive excipients and established mucoadhesive materials were investigated. Some nonadhesive materials showed unexpectedly good adhesive properties when tested as tablets but not as powders. Conversely, some bioadhesive materials had unexpectedly low adhesive properties when tested as tablets. Thus, powder specimens of some materials appear to give more realistic results than tablet specimens in this respect. The use of powder specimens seems particularly applicable for testing dispersible tablets intended for transmucosal absorption. Potential for increasing the bioadhesive properties of coarse, nonadhesive carrier particles by coating them with fine particles of bioadhesive materials during dry mixing (forming interactive mixtures) was also studied. The tensile strength of the adhesive bond between the mucosa and the nonadhesive excipients was improved when fine cross-linked carboxymethyl cellulose sodium (Ac-Di-Sol) particles were added. The addition of increased proportions of Ac-Di-Sol initially improved the bioadhesive properties until a plateau was reached. A standardised test of bioadhesive capacity could therefore involve the addition of fine bioadhesive powders to coarse carriers in proportions close to those providing monoparticulate surface coverage. Interactive mixtures such as these may also offer potential as a tool for use in the development of bioadhesive drug formulations. PMID- 12631409 TI - Erythromycin, an inhibitor of mitoribosomal protein biosynthesis, alters the amphotericin B susceptibility of Candida albicans. AB - Exposure of the yeast Candida albicans to the macrolide antibiotic erythromycin (C(37)H(67)NO(13)) results in elevated tolerance to the polyene antifungal amphotericin B. Erythromycin displays no fungistatic activity against C. albicans but inhibits the synthesis of cytochromes, particularly cytochrome aa(3). Consequently there is a reduction in aerobic respiration by up to 90% when cells are exposed to 10 mg mL(-1) erythromycin. Cellular ergosterol levels are also severely reduced. Erythromycin inhibits protein biosynthesis in ribosomes (mitoribosomes) located within the mitochondrion of the yeast cell, which results in a disruption of cytochrome biosynthesis with an adverse effect on respiration. The synthesis of ergosterol is oxygen dependent and consequently ergosterol levels are depleted in erythromycin-treated C. albicans. Ergosterol is the target for amphotericin B and since there is less of this sterol in erythromycin-treated cells, there is an increase in tolerance of the antifungal agent. Our work indicates that co-administration of erythromycin and amphotericin B to control bacterial and fungal infections, respectively, may inadvertently lead to an elevation in the tolerance of C. albicans for this antifungal agent. PMID- 12631410 TI - Supercritical fluid processing of proteins: lysozyme precipitation from aqueous solution. AB - Aqueous solutions of hen egg lysozyme (3% w/v) were dispersed and precipitated by a homogenous mixture of supercritical carbon dioxide-ethanol using the Solution Enhanced Dispersion by Supercritical fluid (SEDS) process. The effects of different working conditions, such as temperature, pressure and the flow rates of the solution and ethanol, on the particle-formation process were studied. The morphology, particle size and size distribution and biological activity of the protein were determined. The precipitates were examined with high-sensitivity differential scanning calorimetry (HSDSC) and high-performance cation-exchange chromatography. Particle size measurements showed the precipitates to be aggregates with primary particles of size 1-5 microm. The similarity of HSDSC data for unprocessed and processed samples indicated that the different physical forces that stabilise the native form of lysozyme are unchanged after SEDS processing. From FT-Raman spectroscopic studies secondary structural changes were observed in certain SEDS-produced lysozyme, with most processed samples displaying a slightly more disordered secondary structure than the unprocessed sample. However, SEDS samples produced at 200 bar and 40 degrees C exhibited negligible disturbance. Thus the SEDS process utilising aqueous solution was able to bring about size reduction of lysozyme with minimal loss of biological activity. PMID- 12631411 TI - Therapeutic equivalence of a low dose artemisinin formulation in falciparum malaria patients. AB - We have evaluated the therapeutic equivalence of a beta-cyclodextrin-artemisinin complex at an artemisinin dose of 150 mg, with a commercial reference preparation, Artemisinin 250 at a recommended dose of 250 mg. One hundred uncomplicated falciparum malarial patients were randomly assigned to orally receive either beta-cyclodextrin-artemisinin complex (containing 150 mg artemisinin) twice daily for five days or the active comparator (containing 250 mg artemisinin) twice daily for five days. The patients were hospitalized for seven days and were required to attend follow up assessments on days 14, 21, 28 and 35. All patients in both treatment groups were cured of the infection and achieved therapeutic success. At day seven of treatment, all patient blood was clear of the parasites and the sublingual temperature of all patients was less than 37.5 degrees C. Moreover, the parasite clearance time in both treatment groups was similar, being approximately three days after initiation of treatment. Comparable plasma artemisinin concentrations were observed between patients in both treatment groups at 1.5 and 3.0 h, although slightly higher levels were obtained with patients in the beta-cyclodextrin-artemisinin complex-treated group. The beta-cyclodextrin-artemisinin complex at a dose of 150 mg artemisinin was therapeutically equivalent to 250 mg Artemisinin 250. Additionally, patients receiving beta-cyclodextrin-artemisinin complex showed less variability in their plasma artemisinin concentrations at 1.5 h post-dosing, which suggested a more consistent rate of drug absorption. PMID- 12631412 TI - Profound difference in pharmacokinetics between morin and its isomer quercetin in rats. AB - Morin and quercetin are isomeric antioxidant flavonols widely distributed in plant foods and herbs. The pharmacokinetics of both flavonols at two doses were investigated and compared in rats. Parent forms and their glucuronides and sulfates in serum were determined by HPLC before and after enzymatic hydrolysis, respectively. After oral dosing of morin, both the parent form, morin, and its glucuronides and sulfates were present in the bloodstream. The conjugated metabolites predominated at the dose of 25 mg kg(-1), whereas the parent form was predominant at the dose of 50 mg kg(-1). Moreover, the AUC of morin parent form increased by a factor of 37 when the dose doubled, indicating that morin showed nonlinear pharmacokinetics. On the other hand, quercetin presented only as glucuronides and sulfates in the blood, indicating negligible bioavailability of quercetin, and the metabolites showed linear pharmacokinetics at the two doses studied. When considering the total AUC of parent form with conjugated metabolites, the extent of absorption of morin was 3 fold that of quercetin at the dose of 50 mg kg(-1). The results indicated that the difference in hydroxylation pattern on B-ring of flavonol markedly affected their fates in rats. PMID- 12631413 TI - Comparison of metabolic pharmacokinetics of baicalin and baicalein in rats. AB - Baicalin and baicalein, a flavone glucuronide and its aglycone, are bioactive constituents of Scutellariae Radix with various beneficial activities. We have characterized and compared the metabolic pharmacokinetics of baicalin and baicalein in rats. Baicalein was administered intravenously and orally to rats, and baicalin was orally administered. An HPLC method was used to determine the concentration of baicalein before and after hydrolysis using beta glucuronidase/sulfatase. The pharmacokinetic parameters were calculated by using WINNONLIN. Unpaired Student's t-test was used for statistical comparison. The result showed that after intravenous administration of baicalein, 75.7% of the dose was circulating as its conjugated metabolites. After oral administration of baicalein, absorption of baicalein itself was negligible, whereas the glucuronides/sulfates of baicalein were predominant in the plasma. When compared with intravenous bolus administration with dose correction, the absolute absorption was 40%. When baicalin was administered orally, glucuronides and sulfates of baicalein were exclusively circulating in the plasma. The relative absorption for baicalin was 65% when compared with baicalein. Profound differences of serum profile and pharmacokinetics were observed between oral baicalein and baicalin. Baicalin demonstrated significantly later time to peak concentration (t(max)) and lower peak serum concentration (C(max)) of baicalein conjugated metabolites than baicalein, indicating baicalin was absorbed more slowly and to a lesser extent than baicalein. PMID- 12631414 TI - Structure-activity relationship determination within a group of substituted phenyl sulfamate based compounds against the enzyme oestrone sulfatase. AB - The enzyme oestrone sulfatase (ES) is responsible for the conversion of the stored (sulfated) form of oestrogens to the active form, namely oestrone. In our continuing quest to synthesize potent inhibitors of oestrone sulfatase and to determine the structural requirements for such inhibition, we have synthesized and evaluated several derivatives of phenyl sulfamate. We report the results of the synthesis and biochemical evaluation of a series of 3- and 4-aminosulfonated derivatives of phenol in an effort to investigate the role of the acid dissociation constant (pK(a)), and therefore the stability of the phenoxide ion, on the inhibitory activity of compounds against this enzyme. The results showed that there was a strong correlation between the observed pK(a) and inhibitory activity within the aminosulfonated compounds considered. This suggested that in the inhibition of oestrone sulfatase by these compounds, pK(a) was an important physicochemical property, and as such, the stability of the O(-) ion was a crucial factor in the inhibition, and therefore the drug design process. PMID- 12631415 TI - Therapeutic usefulness of Keishi-bukuryo-gan for diabetic nephropathy. AB - Keishi-bukuryo-gan is a traditional herbal medicine, which is used clinically as a vascular system disorder-eliminating drug. In this study, its effect on the progression of diabetic nephropathy in experimental rats was investigated. The diabetic nephropathy model used in this study shows functional and morphological changes of the kidney resembling those seen in patients with diabetic nephropathy. Increased proteinuria and serum urea nitrogen and creatinine levels and decreased creatinine clearance, which are important parameters of renal function, were observed in rats with diabetic nephropathy. Pathological examination of the kidney revealed diffuse, nodular and exudative lesions and arteriolar hyalinosis. The deterioration of renal function was ameliorated in rats treated with Keishi-bukuryo-gan for 15 weeks and these results agreed with the renal histological findings. In addition, metabolic abnormalities mediated by persistent hyperglycaemia (the glycation reaction, excessive polyol pathway activity, oxidative stress and lipid metabolic abnormalities) were also observed. However, Keishi-bukuryo-gan reduced accumulation of advanced glycation end products, determined by measuring fluorescence, and serum lipid peroxidation, triglyceride and total cholesterol levels dose-dependently. Thus, this study indicates the potential therapeutic usefulness of Keishi-bukuryo-gan for retarding the progression of renal damage and suggests that its beneficial effects were due to its ability to improve metabolic abnormalities associated with diabetes. PMID- 12631416 TI - Inhibitory effects of urinary metabolites on platelet aggregation after orally administering Shimotsu-To, a traditional Chinese medicine, to rats. AB - Shimotsu-To, which consists of four herbal extracts, has been used clinically for improving abnormal blood coagulation, fibrinolysis, atherosclerosis and chronic inflammation in Japan and China. We have investigated the pharmacological relationship between the effects and chemical components of Shimotsu-To after oral administration to rats. The urinary constituents were separated and identified by three dimensional (3D-) HPLC equipped with a photodiode array detector as a new tool and the chemical structures were determined by spectroscopic methods to be trans-ferulic acid-3-O-sulfate (1), vanillic acid (2), m-hydroxyphenylpropionic acid (3), trans-ferulic acid (4) and cis-ferulic acid (5). Of these compounds, 2-5 strongly inhibited platelet aggregation induced by ADP and arachidonic acid. Compound 1, the sulfate conjugate of 4, did not show any inhibitory effect, which suggested that the inhibitory effect on platelet aggregation was inactivated by sulfate conjugation. These results indicated that compounds 2-5 partly contributed to the anti-Oketsu effect of Shimotsu-To through the inhibition of platelet aggregation. PMID- 12631418 TI - Sensitivity to beta-adrenoceptor agonists of adipocytes from rats treated with an aqueous extract of Croton cajucara Benth. AB - Aqueous extracts of Croton cajucara bark are used in folk medicine to treat hepatic and gastrointestinal disorders and as a coadjuvant in weight-loss programs. We examined the effect of treating rats for 15 days with a 5% aqueous extract of C. cajucara on body weight and food intake. The epididymal adipose pads were removed and the lipolytic responses of isolated adipocytes to isoprenaline, noradrenaline (norepinephrine), BRL37344 and adrenaline (epinephrine) were analysed in the absence or presence of metoprolol or ICI118,551. Treated rats had a significantly lower weight gain than control rats, with no difference in food and liquid intake, epididymal fat-pad weight or basal glycerol release. The sensitivity of the lipolytic response to isoprenaline and adrenaline was significantly higher in adipocytes from treated rats. The sensitivity to noradrenaline or BRL37344 was unaltered. Metoprolol shifted the dose-response curves to noradrenaline to the right in adipocytes from control and treated rats; the dose-response curve to isoprenaline in adipocytes from control rats was also shifted to the right. In adipocytes from treated rats, the dose response curve to isoprenaline was unaltered by metoprolol but was shifted to the right by ICI118,551, a beta(2)-adrenoceptor antagonist. We conclude that in adipocytes from treated rats there is an increase in the lipolytic response to non-selective agonists (isoprenaline and adrenaline) mediated by beta(2) adrenoceptors, with no alteration in the responses mediated by beta(1) adrenoceptors (noradrenaline) or beta(3)-adrenoceptors (BRL37344). This effect could increase the role of adrenaline as an endogenous stimulator of lipolysis. PMID- 12631417 TI - Inhibition of TPA-induced NF-kappaB nuclear translocation and production of NO and PGE2 by the anti-rheumatic gold compounds. AB - Auranofin, aurothioglucose and aurothiomalate (10 microM each) inhibited 12-O tetradecanoylphorbol 13-acetate (TPA, 16.2 nM)-induced nuclear translocation of nuclear factor-kappa B (NF-kappaB), and production of nitric oxide (NO) and prostaglandin E(2) (PGE(2)) in rat peritoneal macrophages when the cells were pre incubated with each gold compound for 20 h. Without pre-incubation for 20 h, aurothioglucose and aurothiomalate, but not auranofin, failed to inhibit the TPA induced NF-kappaB nuclear translocation and production of NO and PGE(2). Auranofin, aurothioglucose and aurothiomalate did not affect the direct binding of NF-kappaB to the DNA probe. It was suggested that these gold compounds inhibit the TPA-induced production of NO and PGE(2) by inhibiting the NF-kappaB nuclear translocation. PMID- 12631419 TI - Pharmaceutical ethnobotany in the Montseny biosphere reserve (Catalonia, Iberian Peninsula). General results and new or rarely reported medicinal plants. AB - An ethnobotanical survey was carried out in the massif of Montseny, which is situated in north-east Catalonia (Iberian Peninsula), covers 826 km(2) and has a population of 80 000. From 1993 to 2000 we interviewed 180 people and collected ethnobotanical information about 351 medicinal plants, with 4023 use-reports. Through comparison with a large set of studies, we detected 501 unreported or uncommon uses that corresponded to 201 plant species, 57 of which had never or very rarely been cited as medicinal or toxic. General results and key findings on the plant ethnopharmacology (number of medicinal plants, modes of consumption, types of illnesses treated) of this area are presented here, together with information on new or rarely reported medicinal plants. PMID- 12631420 TI - Possible involvement of peripheral serotonin 5-HT3 receptors in fluvoxamine induced emesis in Suncus murinus. AB - Selective serotonin reuptake inhibitors fluvoxamine and fluoxetine, as well as serotonin (5-HT), induced vomiting in Suncus murinus (a house musk shrew). Fluvoxamine- and fluoxetine-induced vomiting gradually decreased with their repeated administration. Vomiting induced by serotonin also decreased with repeated treatment with serotonin. In these shrews, fluvoxamine-induced vomiting was partially inhibited. Fluvoxamine might induce vomiting, at least partially, by indirectly activating peripheral 5-HT(3) receptors, since serotonin has been reported to induce vomiting by activating peripheral 5-HT(3) receptors and granisetron, a 5-HT(3) antagonist, partially suppressed fluvoxamine-induced vomiting in our previous finding. In addition, fluvoxamine-induced vomiting was impaired more effectively using a step-wise dose-up schedule of fluvoxamine than a fixed high-dose schedule. Therefore, a careful dosing strategy starting with a low dose might be effective for avoiding emesis associated with the clinical use of fluvoxamine. PMID- 12631422 TI - Hypoxic ventilatory response, ventilation, gas exchange, and fluid balance in acute mountain sickness. AB - To examine whether sea-level hypoxic ventilatory responses (HVR) predict acute mountain sickness (AMS) and document temporal changes in ventilation, HVR, gas exchange, and fluid balance, we measured these parameters at low altitude (100 m) and daily during 3 days at high altitude (4559 m). At low altitude, there were no significant differences in rest or exercise isocapnic HVR, poikilocapnic HVR at rest, and hypercapnic ventilatory response between 12 subjects without significant AMS and 11 subjects who fell sick. No low altitude ventilatory responses correlated with AMS or fluid balance at high altitude. On day 1, isocapnic HVR was significantly lower in the AMS group [0.86 +/- 0.43 (SD) vs. 1.43 +/- 0.63 L/min/% Sa(O2), p < 0.05). AMS was associated with higher AaD(O2), lower Pa(O2), and Sa(O2), while Pa(CO2) was not different between subjects with and without AMS. Both groups showed equivalent reductions in urine volume, sodium output, and gain in body weight on day 1 while climbing to 4559 m, but on day 2 only subjects without AMS had diuresis, natriuresis, and weight loss. We conclude that (1) susceptibility to AMS, fluid balance, and ventilation at high altitude cannot be predicted by low altitude HVR testing and (2) that the failure to increase HVR on arrival at high altitude and impaired gas exchange, possibly due to interstitial edema, may account for the more severe hypoxemia in AMS. PMID- 12631424 TI - Transient minor improvement of high altitude headache by sumatriptan. AB - High-altitude headache often fulfills the criteria of migraine. Therefore, we hypothesized that sumatriptan, a 5-HT1 receptor agonist specifically effective for treatment of migraine, would also alleviate high altitude headache. A randomized, placebo-controlled double-blind trial was performed on 29 mountaineers with at least moderate headache on the day of arrival at 4559 m. Fourteen subjects received 100 mg sumatriptan orally and 15 subjects received placebo. Before treatment there were no significant differences between groups regarding rate of ascent, duration and severity of headache, and acute mountain sickness score. All 6 female subjects were randomly assigned to placebo. Absolute values and the reduction of headache scores 1, 3, and 12 h after the administration of sumatriptan did not differ between treatment groups, but headache scores tended to be lower with sumatriptan after 1 or 3 h when compared with placebo. Considering only male mountaineers, there was a significant decrease of headache scores after 1 and 3 h. Because there was only a minor transient amelioration of high altitude headache with sumatriptan, we conclude that 5-HT1 receptors do not play a major role in the pathophysiology of high altitude headache. PMID- 12631423 TI - Effect of high altitude on protein metabolism in Bolivian children. AB - In Bolivia, malnutrition in children is a major health problem that may be caused by inadequate protein, energy, and micronutrient intake; exposure to bacterial and parasitic infections; and life in a multistress environment (high altitude, cold, cosmic radiation, low ambient humidity). However, no data on protein absorption and utilization at high altitude were available. Therefore, we evaluated the effect of altitude on protein metabolism in Bolivian children. We measured protein utilization using leucine labeled with a stable isotope ((13)C) in two groups of healthy prepubertal children matched for age. Group 1 (n = 10) was examined at high altitude (HA) in La Paz (3600 m), and group 2 (n = 10) at low altitude (LA) in Santa Cruz (420 m). The nutritional status did not differ between groups but, as was to be expected, the HA group had higher hemoglobin concentration than the LA group. The children consumed casein that was intrinsically labeled with L-(1-(13)C) leucine and expired (13)CO(2) was analyzed. Samples of expired air were measured by isotope ratio mass spectrometer in Clermont-Ferrand. It was found that cumulative leucine oxidation ((13)CO(2)) at 300 min after ingestion was 19.7 +/- 4.9% at HA and 25.2 +/- 3.2% at LA. These results showed that protein absorption and/or utilization is significantly affected by altitude. PMID- 12631426 TI - Highest permanent human habitation. AB - The aim of this analysis was to determine the altitude of the highest permanent human habitation in the hope that this will throw some light on what determines the highest altitude that a community can tolerate indefinitely. A number of places where people have lived at very high altitudes for long periods of time are reviewed. Individuals have lived for as long as 2 yr at an altitude of 5950 m, and there was a miner's camp at 5300 m for several years. The highest permanently inhabited town in the world at the present time appears to be La Rinconada, a mining village of over 7000 people in southern Peru at an altitude of up to 5100 m, which has been in existence for over 40 yr. The altitude of the highest permanent human habitation is determined partly by economic factors, rather than solely by human tolerance to hypoxia. PMID- 12631425 TI - Neuropathic pain and dysesthesia of the feet after Himalayan expeditions. AB - High altitude peripheral nerve disease secondary to frostbite or trauma is a well recognized medical problem during mountaineering expeditions. However, in our experience as medical professionals on 19 expeditions to the Himalayas in the years 1977 to 2000, an unusual syndrome of neuropathic pain and/or dysesthesia in both feet apparently unrelated to frostbite or trench foot was observed in 8 (4.8%) of 165 European mountaineers. Mountaineers complained of persistent and continuous pain, which was consistently described as a "corky" sensation in their feet, associated with severe lancinating exacerbations. Pain improved with cold and worsened with heat and gentle pressure. Symptoms were incapacitating in a third of the cases. Treatment with carbamazepine was effective, and the disorder evolved to total resolution in 4 to 8 weeks. We present the case of a patient who had this syndrome and in whom complete work-up studies done on his arrival home, 14 days after its presentation, were unrevealing. The paucity of information regarding this particular variety of neuropathic pain of the feet may be due to lack of clinical suspicion in the field, favorable outcome, and difficulties for further study and evaluation. PMID- 12631427 TI - A year's experience as advisory doctor to a commercial mountaineering expedition company. AB - The purpose of this paper is to outline one U.K. doctor's experience as advisor to a British commercial high altitude expedition company. Within the United Kingdom, this is pioneering work, and various aspects of the job are outlined that have evolved in response to the current medical needs and market forces. The U.K. medicolegal aspects of the work are considered in the light of there being no current case law to guide doctors in this role. With more people worldwide making use of commercial companies for expeditions with the associated duty of care, it is hoped that this will stimulate further debate on some potentially delicate issues that cross international boundaries. PMID- 12631428 TI - Acute mountain sickness. PMID- 12631429 TI - On-site treatment of avalanche victims ICAR-MEDCOM-recommendation. PMID- 12631430 TI - Stress hormones in the research on cardiovascular effects of noise. AB - In recent years, the measurement of stress hormones including adrenaline, noradrenaline and cortisol has been widely used to study the possible increase in cardiovascular risk of noise exposed subjects. Since endocrine changes manifesting in physiological disorders come first in the chain of cause-effect for perceived noise stress, noise effects in stress hormones may therefore be detected in populations after relatively short periods of noise exposure. This makes stress hormones a useful stress indicator, but regarding a risk assessment, the interpretation of endocrine noise effects is often a qualitative one rather than a quantitative one. Stress hormones can be used in noise studies to study mechanisms of physiological reactions to noise and to identify vulnerable groups. A review is given about findings in stress hormones from laboratory, occupational and environmental studies. PMID- 12631431 TI - Inner ear damage in children due to noise exposure from toy cap pistols and firecrackers: a retrospective review of 53 cases. AB - This retrospective study presents the findings of inner ear damage documented in 53 children exposed to impulsive sound emitted by toy weapons and firecrackers. There were 49 boys and four girls aged between four and fourteen years. Thirty nine children were affected unilaterally while fourteen had bilateral hearing loss (total of 67 ears). Most of the hearing loss (>70%) was sensorineural high frequency hearing loss, while only nine out of the 67 injured ears had sensorineural mid frequency hearing loss. Seven children sustained a traumatic ear drum perforation. Dizziness or tinnitus was reported by twenty children, with pathological ENG findings in four of them. This paper re-emphasizes the possibility of inner ear damage in children from exposure to noisy toys. PMID- 12631432 TI - The work of I-INCE Technical Study Group 2 on noise labels for consumer and industrial products. AB - In 1999 a new I-INCE Technical Study Group TSG 2 was formed on "Noise labels for consumer and industrial products". This was intended to survey current methods for labelling and otherwise characterizing the noise emissions of consumer and industrial products. Note that labelling can mean more than just a physical label - it might be details in a Technical Manual. The measurement methods used by testing authorities were to be included in the survey. The methodologies were to be compared, and an assessment made of their relative effectiveness. The study of noise labelling is part of an educational program to advise on how, and in what form such labelling should be implemented. There has been active participation in the TSG from UK, USA, Japan, Norway, Turkey, Belgium and Brazil, with email exchange of information and 3 meetings, at Internoise 2000 in Nice, 2001 in Den Haag and 2002 in Dearborn, USA. More recently the survey questionnaire has been sent to all the 46 Member Societies of I-INCE. This paper explains the survey and summarises current results. PMID- 12631433 TI - Does health promotion work in relation to noise? AB - Noise is a health risk. The only scientifically established adverse health effect of noise is noise-induced hearing loss (NIHL). Besides noise may affect quality of life and cause annoyance and sleep disturbance. The present scientific evidence of potential non-auditory effects of noise on health is quite weak. Whether health promotion works in relation to noise may be reflected by permanent hearing threshold shift development in population studies. Hearing impairment continues to be the most prevalent disability in Western societies. The National Institute of Occupational Safety and Health (NIOSH) still rates noise induced hearing loss among the top ten work-related problems. Recent studies report that employees continue to develop noise induced hearing loss although to a lesser extent than before, in spite of occupational hearing conservation programmes. Besides socio-acusis and leisure noise seem to be an increasing hazard to hearing, also in young children and adolescents. This seems partly related to acute leisure noise exposure (e.g. toy pistols, amplified music). However, population studies increasingly find non-normal high-frequency hearing including the characteristic NIHL-"notch" around 6 kHz also in subjects who do not report noise exposure incidents or activities. Today 12.5% of US children 6-19 years show a noise-"notch" in one or both ears (n= 5249, Niskar et al 2001). A Norwegian county audiometry survey on adults >/= 20 years n=51.975) showed mean unscreened thresholds +10 dB at 6 kHz for both genders even or the youngest age group 20-24 years (Borchgrevink et al 2001). Accordingly, the present health promotion initiatives seem insufficient in relation to noise and noise-induced hearing loss. PMID- 12631434 TI - Do public inquiries for noise control serve a useful purpose?--An acoustic consultant's view. AB - In the United Kingdom, before the introduction of the various town and country planning acts and associated regulations, landowners were free to use their land in any way they wished, subject only to limitations imposed by lease or covenant and the avoidance of nuisance or trespass against neighbours. Any disputes arising would be resolved by negotiation or via a court of law. Under current planning laws and regulations, local authorities are empowered to impose special conditions or even to refuse development to prevent excessive nuisance, but the resulting noise management solutions are not always optimum from either the noise maker's or the noise exposed's points of view. In addition, the planning system has almost no effect on existing noise. Public inquiries provide a useful mechanism for the investigation of appeals against local authority decisions, or where the government has decided that issues of strategic or national importance need to be fully explored in a public forum. In practice, and largely because of individual disagreement, public inquiries can result in excessive delays while all interested parties are allowed to have their say. There seems to be an increasing consensus that the general inadequacy of existing methods of assessing noise impact is at least partly to blame. The new European Environmental Noise Directive represents a step change towards the imposition of one-size-fits-all regulatory or administrative procedures which should eventually contribute towards the reduction of public inquiry delays, but on the other hand, any weakening of the general principle of basing decisions on 'informed flexibility' will probably have significant negative consequences over the longer term. PMID- 12631435 TI - Ambient noise strategy: a solution for noise control? AB - The British Government earlier this year undertook a consultation on its proposal, announced in the Rural White Paper, to develop an Ambient Noise Strategy in England. The proposals envisage a three phase approach: In phase 1 we would aim to establish three key sets of information: information on the ambient noise climate in the country--i.e. the number of people affected by different levels of noise, the source of that noise (road, rail, airports and industry) and the location of the people affected, by producing noise maps of the main sources of noise; methods which the Government might use to assess the effects of noise- particularly regarding people's quality of life and tranquility; the techniques available to take action to improve the situation where bad or preserve it where good. In phase 2 we would aim to evaluate and identify options for prioritising the various alternatives from phase 1 in terms not only of costs and benefits but also time-scales and synergies and conflicts with other Government priorities including economic and social issues. An optimal policy reduces noise at lowest net cost, whilst capturing as many synergistic benefits, and minimising any potentially adverse impacts. Decision makers need to ensure that the impacts of the noise policies do not cost society more than the benefits expected. A recent study undertaken by the Government, looked at how a cost-benefit type framework could be used, with noise maps, to help inform such decisions. Finally, in phase 3, the Government would need to agree on the necessary policies to move towards the desired outcome--i.e. the National Ambient Noise Strategy itself. The results of the consultation are expected to be published later this year. PMID- 12631436 TI - Noise control in the transportation corridor. AB - This paper considers the opportunities for noise control within the route corridor required for construction of road, rail and other guided transport schemes. It deals with control of noise generation at source, and in the transmission path close to the point of generation. In this way it is possible to control the amount of acoustic power generated, and to absorb part of the radiated power at points of reflection. Purely reflective wayside barriers do little to absorb acoustic energy, merely reflecting it in a different direction. Whilst this has selfish benefits to the receptor in the shadow zone of the barrier, it makes things worse for others on the reflective side of the geometry. The paper therefore considers the options available to the engineer in the design of rolling and sliding interfaces and the use of acoustically absorptive finishes on all surfaces close to the point of noise generation. This includes the running surface itself, structural components, retaining walls, over and under passes, and the inner surfaces of track and wayside barriers. PMID- 12631437 TI - The role of hearing protectors in leisure noise. AB - Ear muffs and ear plugs are widely used in the workplace to provide hearing protection for employees exposed to high levels of noise. Through an examination of the use of ear protectors in the workplace this paper explores the extent to which these devices can play a similar role in protecting members of the public from hearing damage arising from exposure to high levels of noise from leisure activities. It is concluded that the major limitation to the effective use of ear protectors for leisure use is likely to be the lack of easily accessible information, advice and guidance on the nature of the hearing protection risk from noisy leisure activities, and on the availability, selection and use of protectors, and of the need for regular hearing checks. PMID- 12631438 TI - The concept of noise sensitivity: implications for noise control. AB - The term "noise sensitivity" is frequently used in many areas of noise research. However, it can be used to describe several different effects and it can be measured in different ways. In noise surveys, noise sensitivity refers to the fact that individuals differ in the annoyance produced by different sources of noise. Noise sensitivity can be viewed as an independent variable, which may be directly related to outcomes such as health status, or it can be conceptualized as a factor that modifies or mediates the effects of noise exposure on the outcome measure. Noise sensitivity is highly correlated with the general trait negative affectivity, a measure of the extent to which individuals perceive or report negative features of their environment or self. Indeed, few studies have demonstrated effects of noise sensitivity that are independent of negative affectivity. This implies that it is most appropriate to examine general indicators of reported sensitivity rather than a noise-specific measure. Noise sensitivity can also be considered in terms of physiological reactivity to noise sources. Such effects are often only weakly associated with self-reports of noise sensitivity. Habituation to noise is also an important topic to consider and again this appears to be largely independent of self-reported noise sensitivity. Overall, it would appear that it is important to distinguish between subjective reports of noise sensitivity and objective indicators. Different factors will modify these two aspects of noise sensitivity and this implies that different strategies are needed to influence them. Such effects must be taken into consideration when one considers whether control should be targeted at the community in general, or whether it should also cover the most sensitive individuals. PMID- 12631439 TI - Transplasma membrane electron transport: enzymes involved and biological function. AB - The notion of transmembrane electron transport is usually associated with mitochondria and chloroplasts. However, since the early 1970s, it has been known that this phenomenon also occurs at the level of the plasma membrane. Ever since, evidence has accumulated for the existence of a plethora of transplasma membrane electron transport enzymes. In this review, we discuss the various enzymes known, their molecular characteristics and their biological functions. PMID- 12631440 TI - Oxidative stress in ataxia telangiectasia. AB - Ataxia telangiectasia is one of a group of recessive hereditary genomic instability disorders and is characterized by progressive neurodegeneration, immunodeficiency and cancer susceptibility. Heterozygotes for the mutated gene are more susceptible to cancer and to ischaemic heart disease. The affected gene, ATM (ataxia telangiectasia mutated), has been cloned and codes for a protein kinase (ATM), which orchestrates the cellular response to DNA double-strand breaks after ionising radiation. An underlying feature of ataxia telangiectasia is oxidative stress and there is chronic activation of stress response pathways in tissues showing pathology such as the cerebellum, but not in the cerebrum or liver. ATM has also been shown to be activated by insulin and to have a wider role in signal transduction and cell growth. Many, but not all, aspects of the phenotype can be attributed to a defective DNA damage response. The oxidative stress may result directly from accumulated DNA damage in affected tissues or ATM may have an additional role in sensing/modulating redox homeostasis. The basis for the observed tissue specificity of the oxidative damage in ataxia telangiectasia is not clear. PMID- 12631441 TI - Assessment of membrane protection by traditional Chinese medicines using a flow cytometric technique: preliminary findings. AB - In this preliminary study, we used a 'living cell' flow-cytometric approach to membrane protection by four traditional Chinese medicines (TCMs). Cells were incubated, separately, for 30 min with aqueous extracts (1.5% w/v) of lingzhi (Ganoderma lucidum), ginger (Zingiber officianale), ginseng (Panax ginseng), and green tea (Camellia sinensis). Membranes were labelled with a fluorescent probe, cells were then incubated with cumene hydroperoxide, and site-specific oxidation induced by iron/ascorbate. Oxidation of membrane lipids quenches fluorescence. Forward-scatter fluorescence was measured at timed intervals after initiation of oxidation. Results indicate that lingzhi and ginger contain antioxidant component(s) that act within the cell membrane and slow lipid peroxidation in situ. Results demonstrate also that this living cell model is a useful biomonitoring tool to help determine molecular aspects of putative health effects of TCMs. PMID- 12631442 TI - Ionotropic glutamate receptors are involved in malondialdehyde production in anesthetized rat brain cortex: a microdialysis study. AB - Elevated extracellular glutamate levels can increase malondialdehyde production in the brains of anesthetized rats. Thus, we investigated whether ionotropic glutamate receptors are involved in glutamate-induced malondialdehyde production. A microdialysis probe was implanted in the brain cortex of anesthetized rats. The malondialdehyde level in microdialysates was analyzed using an HPLC system. Three different ionotropic glutamate receptor agonists were used. At a concentration of 1.5 mM alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid hydrobromide (AMPA, a selective AMPA receptor agonist) induced a dramatic increase in extracellular malondialdehyde production (as much as 14-fold relative to the basal value). N-Methyl-D-aspartic acid (NMDA, a selective NMDA receptor agonist) also induced an increase in extracellular malondialdehyde production; however, the increase was not as much as that observed in the perfusion of AMPA receptor agonist. Kainic acid (a selective kainate receptor agonist) did not significantly increase malondialdehyde production. When co-perfused with L-trans-pyrrolidine 2,4-dicarboxylate (PDC; 31.4 mM), a glutamate uptake transport inhibitor that can increase the extracellular glutamate levels, AMPA receptor antagonist [1-(4 aminophenyl)4-methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine hydrochloride, 1.0 mM] can significantly reduce PDC-induced malondialdehyde production. Although NMDA receptor antagonist [(5R,10S)-(+)-5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate, MK801] also can decrease the PDC-induced malondialdehyde production, it was not as effective as the AMPA receptor antagonist. These results suggest that ionotropic receptors are involved in the glutamate-induced increase in malondialdehdye production. Specifically, AMPA receptor seems to be predominant in the glutamate-induced malondialdehdye production in anesthetized rat brain cortex. PMID- 12631444 TI - Protection against cisplatin-induced nephrotoxicity by Cu(II)2(3,5 diisopropylsalicylate)4. AB - The ability of Cu(II)(2)(3,5-diisopropylsalicylate)(4), CuDIPS, which exhibits superoxide dismutase (SOD)-like activity, to prevent cisplatin-induced nephrotoxicity was examined in rats. Rats were divided into four groups and treated as follows: (i) vehicle control; (ii) cisplatin (16 mg/kg, intraperitoneally); (iii) CuDIPS (10 mg/kg, intraperitoneally); and (iv) cisplatin plus CuDIPS. Rats were sacrificed 3 days post-treatment. Cisplatin alone resulted in significantly increased plasma creatinine and urea. Administration of 10 mg/kg CuDIPS prevented the cisplatin-induced elevation of plasma creatinine and urea and protected against kidney damage. Relative to controls, rats that received cisplatin treatment displayed a decrease of reduced glutathione (GSH) and elevated platinum and thiobarbituric acid reactive substances (TBARS) levels in the kidney. In comparison with controls, activities of antioxidant enzymes (SOD, CAT, GSH-Px and GSH-Rd) were also reduced in the kidney of rats treated with cisplatin. Administration of 10 mg/kg CuDIPS prevented cisplatin-induced alterations in renal platinum, GSH, TBARS, and antioxidant enzyme activities. This study suggests that the protection offered by CuDIPS against cisplatin-induced nephrotoxicity is partly related to maintenance of renal antioxidant systems. PMID- 12631443 TI - The effect of supplementation with an antioxidant preparation on LDL-oxidation is determined by haptoglobin polymorphism. AB - The genetic polymorphism of haptoglobin (Hp) is an independent risk factor in the pathogenesis of atherosclerosis, a condition in which decreased resistance to in vitro oxidation of LDL-cholesterol is observed. We hypothesised that the Hp polymorphism is one of the factors modulating the resistance to Cu(2+)-induced oxidation of LDL during antioxidant supplementation. In this study, 74 middle aged subjects with increased oxidative stress were allocated to either matched placebo or oral antioxidative treatment (Quatral) once daily for 16 weeks. Study parameters were increase of lag phase (DeltaLAG) and the ratio of lag phase during treatment period versus baseline (relative oxidation resistance, ROR), measured by Cu(2+)-induced oxidation of isolated LDL. Hp phenotypes were determined by starch gel electrophoresis. A significant and persistent increase of DeltaLAG (P < 0.05) and ROR (P < 0.01) were observed after 16 weeks of active treatment versus placebo. Interindividual differences in both parameters were significantly associated with the Hp polymorphism: in the active treatment group, DeltaLAG and ROR were significantly higher in Hp 1-1 subjects (P < 0.01) compared to Hp 2-1 and 2-2. Our data demonstrate that Hp phenotype is one of the modulating factors determining the increased resistance to Cu(2+)-induced oxidation of LDL during antioxidative treatment. PMID- 12631445 TI - Thermosensitive phenotype of Escherichia coli mutant lacking NADP+-dependent isocitrate dehydrogenase. AB - Heat shock may increase oxidative stress due to increased production of reactive oxygen species and/or the promotion of cellular oxidation events. NADP(+) dependent isocitrate dehydrogenase (ICDH) in Escherichia coli produces NADPH, an essential reducing equivalent for the antioxidant system. The protective role of ICDH against heat shock in E. coli was investigated in wild-type and ICDH deficient strains. Upon exposure to heat shock, the viability was lower and the protein oxidation was higher in mutant cells as compared to wild-type cells. Induction and inactivation of antioxidant enzymes were observed after their exposure to heat shock both in wild-type and in mutant cells. However, wild-type cells maintained significantly higher activities of antioxidant enzymes than did mutant cells. These results suggest that ICDH plays an important role as an antioxidant enzyme in cellular defense against heat shock through the removal of reactive oxygen species as well as in the protection of other antioxidant enzymes. PMID- 12631447 TI - The enormous burden of digestive diseases on our healthcare system. PMID- 12631446 TI - Redox regulation of homocysteine-dependent glutathione synthesis. AB - In certain tissues, glutathione biosynthesis is connected to methionine metabolism via the trans-sulfuration pathway. The latter condenses homocysteine and serine to cystathionine in a reaction catalyzed by cystathionine beta synthase followed by cleavage of cystathionine to cysteine and alpha ketoglutarate by gamma-cystathionase. Cysteine is the limiting amino acid in glutathione biosynthesis, and studies in our laboratory have shown that approximately 50% of the cysteine in glutathione is derived from homocysteine in human liver cells. In this study, we have examined the effect of pro- and antioxidants on the flux of homocysteine through the trans-sulfuration pathway in the human hepatoma cell line, HepG2. Our studies reveal that pyrrolidine dithiocarbamate and butylated hydroxyanisole enhance the flux of homocysteine through the trans-sulfuration pathway as has been observed previously with the pro-oxidants, H(2)O(2) and tertiary butyl hydroperoxide. In contrast, antioxidants such as catalase, superoxide dismutase and a water-soluble derivative of vitamin E elicit the opposite effect and result in diminished flux of homocysteine through the trans-sulfuration pathway. These studies provide the first evidence for the reciprocal sensitivity of the trans-sulfuration pathway to pro- and antioxidants, and demonstrate that the upstream half of the glutathione biosynthetic pathway (i.e. leading to cysteine biosynthesis) is redox sensitive as is the regulation of the well-studied enzymes in the downstream half (leading from cysteine to glutathione), namely, gamma-glutamyl-cysteine ligase and glutathione synthetase. PMID- 12631448 TI - Nutritional support for acute pancreatitis. PMID- 12631449 TI - Genetic factors in pancreatitis. AB - A number of genetic mutations have recently been identified that appear to be important in the development of pancreatitis. Point mutations in the cationic trypsinogen gene are capable of initiating pancreatitis. These mutations also provide important insights into the pathophysiology of acute pancreatitis and into potential connections between acute and chronic pancreatitis. Mutations in the genes encoding for the pancreatic secretory trypsin inhibitor and the cystic fibrosis transmembrane conductance regulator more likely work in concert with other genes and environmental factors in affecting disease susceptibility. Although the subject so far has received only a limited amount of study, genetic polymorphisms in a wide range of genes relating to pancreatic function and to regulation of inflammation are likely to play major roles in determining each individual's susceptibility to developing pancreatitis, and its severity if it does develop. PMID- 12631451 TI - Staging strategies for pancreatic adenocarcinoma: what the surgeon really wants to know. AB - Pancreatic cancer remains a highly lethal disease in spite of new developments in early diagnosis, marked improvements in surgical morbidity and mortality, and introduction of promising adjuvant and neoadjuvant therapies. This ongoing poor prognosis can be explained in part by the relatively advanced stage of these malignancies at the time of presentation and diagnosis. The staging process in this disease lacks standardization and differs considerably from center to center. Acknowledging that it is impossible to achieve a perfect workup to rule out micrometastatic disease, controversy persists regarding the optimal initial approach and subsequent staging of these tumors. A better, or at least a more standardized staging system, should help to improve disappointing results in the surgical treatment of pancreatic cancer, although such an improvement will be based more on the selection of patients than the actual therapeutic intervention. This paper addresses the current controversy in staging methodology for pancreatic and periampullary malignancies and offers an approach to the staging of pancreatic cancer. PMID- 12631450 TI - Medical therapy for chronic pancreatitis pain. AB - Chronic pancreatitis should be considered in all patients with unexplained abdominal pain. Management of abdominal pain in these patients continues to pose a formidable challenge. The importance of small duct disease without radiographic abnormalities is now a well-established concept. It is meaningful to determine whether patients with chronic pancreatitis have small duct or large duct disease because this distinction has therapeutic implications. Diagnostic evaluation should begin with simple noninvasive and inexpensive tests like serum trypsinogen and fecal elastase, to be followed where appropriate by more complicated measures such as the secretin hormone stimulation test, especially in patients with suspected small duct disease. No universal causal treatment is available. Non enteric-coated enzyme preparations are useful for treatment of pain, whereas enteric-coated enzyme preparations are preferred for steatorrhea. Octreotide is used increasingly for abdominal pain that is unresponsive to pancreatic enzyme therapy. When medical therapy for chronic pancreatitis pain has failed, endoscopic therapy, endoscopic ultrasound-guided celiac plexus block, and thoracoscopic splanchnicectomy, performed by experts, may be considered for a highly selected patient population. Surgical ductal decompression is appropriate in patients with considerable pancreatic ductal dilation. The role and efficacy of cholecystokinin-receptor antagonists, antioxidants, and antidepressant drugs remain to be defined. PMID- 12631452 TI - Gallstone pancreatitis: when is endoscopic retrograde cholangiopancreatography truly necessary? AB - Acute pancreatitis is an inflammation of the pancreas that can, in a minority of patients, lead to local complications, multiorgan failure, and death. Gallstones are the most common cause of acute pancreatitis in Western countries. The majority of patients with acute gallstone pancreatitis have mild disease and recover within 3 to 5 days with bed rest and intravenous fluid replacement. In up to 20% of patients, severe pancreatitis develops and can involve pancreatic tissue necrosis and multiorgan failure. Recent advances in the care of patients with gallstone-induced pancreatitis include better severity stratification on hospital admission, more aggressive fluid resuscitation in the early disease course, early use of antibiotics in patients with pancreatic necrosis, a shift from parenteral to enteral feeding regimens, a better defined and less aggressive approach to pancreatic surgery, and the possibility to remove impacted gallstones endoscopically. Urgent endoscopic retrograde cholangiopancreatography and sphincterotomy are recommended in patients with signs of cholangitis or jaundice, ultrasound evidence of dilated common bile duct, or evidence of severe disease. PMID- 12631453 TI - Intraductal papillary mucinous tumors of the pancreas. AB - Cystic neoplasms of the exocrine pancreas are a small fraction of pancreatic tumors. Within that group of cystic neoplasms, intraductal papillary mucinous tumors (IPMTs) can be distinguished from mucinous cystic neoplasms, serous cystic neoplasms, and pseudopapillary cystic tumors. Awareness of IPMTs has increased since the World Health Organization classified these tumors as its own group in 1996. Because of their favorable prognosis, an extensive diagnostic workup for IPMTs should be performed in patients presenting with cystic lesions of the pancreas. This workup often leads to the diagnosis and the predominant tumor location and size, although the extent of the ductal changes can only be established by histopathology. Surgical resection is the therapy of choice for IPMTs. The type of resection depends upon the extent of the quantitative and qualitative ductal involvement. Total pancreatectomy is currently the treatment for an IPMT that comprises the entire main duct. PMID- 12631454 TI - Mechanisms of acute cholelithiasis. PMID- 12631455 TI - Noninvasive imaging for diagnosis of sphincter of Oddi dysfunction. PMID- 12631456 TI - Understanding risk factors and avoiding complications with endoscopic retrograde cholangiopancreatography. AB - Complications and technical failures of endoscopic retrograde cholangiopancreatography (ERCP) cause significant morbidity and, occasionally, mortality. An understanding of patient- and procedure-related risks is important for decision making with regard to whether or how ERCP should be performed. Instances in which ERCP is the least clearly indicated are often the most likely to cause complications. Patient-related risk factors include suspected sphincter of Oddi (SO) dysfunction, female sex, normal serum bilirubin, or previous history of post-ERCP pancreatitis, with multiple risk factors conferring especially high risk. Technique-related risk factors include difficult cannulation, pancreatic contrast injection, balloon sphincter dilation, and precut sphincterotomy performed by endoscopists of varied experience. Pancreatic stents may reduce the risk of pancreatitis in a number of settings including SO dysfunction. Hemorrhage and perforation are rare and can be avoided with endoscopic technique and attention to the patient's coagulation status. Cholangitis is avoidable with adequate biliary drainage. Because success rates are higher and complication rates lower for endoscopists performing large volumes of ERCP, ERCP should be concentrated as much as possible among endoscopists with adequate experience. Patients with a high risk for complications may be best served by referral to an advanced center. PMID- 12631457 TI - Effects of bioactive agents on biliary motor function. AB - Our understanding of biliary motility under normal and pathophysiologic conditions is still incomplete, but there have been recent advances. Of particular interest are the mechanisms involved in gallbladder filling and emptying, with a focus on understanding the processes underlying impaired gallbladder emptying leading to gallbladder dyskinesia and the formation of gallstones or cholecystitis. The sphincter of Oddi (SO) is a complex neuromuscular structure. Recent studies have attempted to unravel the specific neural or hormonal mechanisms operating under normal physiologic conditions and those that may lead to SO dysfunction. Furthermore, new research fronts are emerging, including the role of leptin in obese patients with impaired biliary motility and the action of electroacupuncture for possible treatment of SO dysfunction. This review illustrates the broad front of current research regarding the effects of bioactive agents on biliary motility, including enteric hormones, nitric oxide, opioids, inflammatory mediators, leptin, protease inhibitors, neurotransmitters, and electroacupuncture. PMID- 12631458 TI - The relationship of bile duct crystals to sphincter of Oddi dysfunction. AB - Bile duct crystals collected either from the duodenum after gallbladder contraction or directly from bile duct aspiration are surrogates for gallbladder stones and microlithiasis. Whether bile crystals also serve as surrogates for bile duct stones or microlithiasis that forms in the bile duct after cholecystectomy is not known based on current data. Sphincter of Oddi dysfunction (SOD), due either to muscular spasm or sphincter fibrosis, is a cause of bile duct obstruction. Almost all of the literature on SOD involves patients who have had a prior cholecystectomy. Intuitively, obstruction at the SO following cholecystectomy would seem to lead to biliary stasis and predispose patients to bile duct microlithiasis. However, a recent study did not find bile duct crystals in patients with manometrically diagnosed SOD. The reason for this is unknown, although we hypothesize that cholesterol and bilirubinate crystals are not surrogates for brown pigment stones commonly found in patients following cholecystectomy. PMID- 12631460 TI - Tumors of the ampulla of Vater. AB - Ampullary tumors are relatively rare, but the occurrence of biliary obstruction early in the disease course facilitates diagnosis. With technologic advances, methods of diagnosis, treatment, and management of ampullary tumors are constantly evolving. However, despite rapid improvements in these areas, preoperative differentiation between adenomas and adenocarcinomas remains difficult. Forcep biopsy specimens can accurately detect the presence or absence of adenomatous changes, but they have a high false-negative rate for adenocarcinoma. Whereas it is generally agreed that all ampullary tumors should be removed or resected, patient selection for the various treatments, including pancreatoduodenectomy, local resection, and endoscopic treatment, remains controversial. Stage of disease, patient characteristics (ie, age and comorbid conditions), and local availability of expertise determine treatment options. PMID- 12631459 TI - Clinical significance of sphincter of Oddi dyskinesia. AB - Sphincter of Oddi dyskinesia (SOD) is a functional disorder of the papilla region that can lead to clinical symptoms and functional obstruction of biliary and pancreatic outflow. Based on the severity of the clinical symptoms, the disorder is classified as one of three types (biliary or pancreatic type I-III). Diagnosis of SOD is hampered by the relative risk of endoscopic sphincter manometry to cause pancreatitis. Manometrically, SOD is characterized by increased pressure in the biliary or pancreatic sphincter segment and can be treated with endoscopic papillotomy. This review is an attempt to balance the arguments for invasive diagnosis with a pragmatic clinical approach in which papillotomy is performed if clinical suspicion and patient presentation support a dysfunction of the papilla. For patients with biliary or pancreatic type I, endoscopic papillotomy is the treatment of choice. In biliary type II, SO manometry may be helpful for clinical decision making; however, the ratio of risks to benefits is difficult to assess based on the present data. In type III SOD, patient selection and the low predictive value of manometry for treatment success raise questions about the clinical usefulness of SO manometry. PMID- 12631462 TI - Cerebral responses following stimulation of unmyelinated C-fibers in humans: electro- and magneto-encephalographic study. AB - There are two kinds of pain, a sharp pain ascending through Adelta fibers (first pain) and a second burning pain ascending though C fibers (second pain). By using a novel method, the application of a low intensity CO(2) laser beam to a tiny area of skin using a very thin aluminum plate with numerous tiny holes as a spatial filter, we succeeded in selectively stimulating unmyelinated C fibers of the skin in humans, and could record consistent and clear brain responses using electroencephalography (EEG) and magnetoencephalography (MEG). The conduction velocity (CV) of the C fibers of the peripheral nerve and spinal cord, probably spinothalamic tract (STT), is approximately 1-4 m/s, which is significantly slower than that of Adelta (approximately 10-15 m/s) and Abeta fibers (approximately 50-70 m/s). This method should be very useful for clinical application. Following C fiber stimulation, primary and secondary somatosensory cortices (SI and SII) are simultaneously activated in the cerebral hemisphere contralateral to the stimulation, and then, SII in the hemisphere ipsilateral to the stimulation is activated. These early responses are easily detected by MEG. Then, probably limbic systems such as insula and cingulate cortex are activated, and those activities reflected in EEG components. Investigations of the cortical processing in pain perception including both first and second pain should provide a better understanding of pain perception and, therefore, contribute to pain relief in clinical medicine. PMID- 12631463 TI - Long-lasting delayed hyperalgesia after chronic restraint stress in rats-effect of morphine administration. AB - Different effects upon the nociceptive response have been observed with exposure to acute and chronic stress in rats. In the present study we repeatedly submitted rats to restraint for 40 days, inducing hyperalgesia using the tail-flick test. A new session of acute stress was applied at the end of 40 days period, and the chronically-stressed animals demonstrated analgesia after forced swimming, but not after restraint. The effect of stress interruption for 14 or 28 days on the nociceptive threshold was then investigated. The basal tail-flick latency remained decreased for at least 28 days (hyperalgesic effect). Following the periods of suspension, the animals were submitted to new session of acute restraint, and stress-induced analgesia was observed only after 28 days of stress interruption. Thus, the mechanisms involved in the long-lasting hyperalgesia presented in this study are not exactly the same as those responsible for the analgesia induced by acute stressors. After 40 days of chronic stress treatment, morphine was injected i.p. (1.0, 5.0 mg/kg or saline). The repeatedly stressed rats displayed decreased morphine effects on nociception compared to unstressed controls. The tolerance of the response to morphine agrees with previous studies suggesting that chronic restraint stress could modify the activity of opioid systems. PMID- 12631461 TI - Proteins interactions implicated in AMPA receptor trafficking: a clear destination and an improving route map. AB - The mechanisms that regulate alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor (AMPAR), synthesis, transport, targeting and surface expression are of fundamental importance to understand the molecular basis of fast excitatory neurotransmission and synaptic plasticity in the mammalian CNS. An area of intense current interest is how AMPARs are directed to the correct locations in the neuron as and when required. This is a multi-layered problem, which involves complex spatio-temporal coordination of multiple protein interactions. Considerable progress has been achieved in identifying a number of proteins that bind directly to AMPAR subunits and the functional consequences of blocking some of these interactions have been determined. This review highlights recent developments in the field. PMID- 12631464 TI - Antisense in vivo knockdown of synaptotagmin I and synapsin I by HVJ-liposome mediated gene transfer modulates ischemic injury of hippocampus in opposing ways. AB - Neurotransmitter release during and after ischemic event is thought to be involved in excitotoxicity as a pathogenesis for the ischemic brain damage, which is mediated by excessive activation of glutamate receptors and attendant calcium overload. To ascertain the role of transmitter release from nerve terminals in promoting the ischemic neurodegeneration, we delivered antisense oligodeoxynucleotides (ODNs) to synaptotagmin I or synapsin I into the rat brain by using HVJ-liposome gene transfer technique. The antisense ODNs were injected into the lateralventricle in rats 4 days prior to transient forebrain ischemia of 20 min. With a single antisense treatment, long-lasting downregulation of the transmitter release relating protein levels at overall synaptic terminals was achieved. The antisense in vivo knockdown of synaptotagmin I prevented almost completely the ischemic damage of hippocampal CA1 neurons, while the in vivo knockdown of synapsin I markedly promoted the ischemic damage of CA1 pyramidal neurons and extended the injury to relatively resistant CA2/CA3 region. The modulation of ischemic hippocampal damage by the in vivo knockdown of synaptotagmin I or synapsin I suggests that transmitter release from terminals plays an important role in the evolution of ischemic brain damage and therefore the transmitter release strategy by the use of antisense ODNs-HVJ-liposome complex is reliable for neuroprotective therapies. PMID- 12631465 TI - Influence of peripheral inflammation on growth-associated phosphoprotein (GAP-43) expression in dorsal root ganglia and on nerve recovery after crush injury. AB - An experimental study was performed to investigate the influence of the inflammation in peripheral target tissue on growth-associated phosphoprotein (GAP 43) expression in dorsal root ganglia (DRG) and on recovery of crushed nerve. Fifty-four male Wistar rats were used for this study. The sciatic nerve was operatively crushed unilaterally with an aneurysm clip. Inflammation in peripheral target tissue was induced by injection of complete Freund's adjuvant (CFA) at 1 week before crush. In crushed or crushed with arthritis rats DRGs were examined in immunohistochemistry for GAP-43 and the sciatic nerves were observed in Epon embedded sections with toluidine blue stain. In addition, electrophysiological studies of the nerves were performed to evaluate the recovery of function. Immunohistochemical studies showed the ratio of GAP-43 immunopositive cells in crushed with arthritis rats was significantly lower than that in crushed rats at 1 week after crush (P<0.01). Electrophysiological studies at 4 weeks after crush showed functional nerve recovery in crushed with arthritis rats was significantly suppressed compared with that in crushed rats (P<0.01). Histological studies showed the mean diameter of the axons in crushed with arthritis rats was significantly smaller than that in crushed rats (P<0.01). All these findings indicate that inflammation in peripheral target tissue suppresses GAP-43 expression in DRG and eventually suppresses functional and morphological recovery of the crushed nerve. PMID- 12631466 TI - Osmotic stress increases cullin-5 (cul-5) mRNA in the rat cerebral cortex, hypothalamus and kidney. AB - Cullin-5 (cul-5), a member of the cullin gene family, may have a role in proteolysis and cell cycle regulation. Our recent study demonstrated that cul-5 mRNA is ubiquitously expressed in the central nervous system and many peripheral organs. The present study used quantitative realtime polymerase chain reaction to measure changes in cul-5 mRNA expression as a consequence of osmotic stress in vivo. Cul-5 mRNA levels were significantly increased in the rat cerebral cortex, hypothalamus and kidney following 48 h of water deprivation. Water deprivation for a shorter time period (24 h) or rehydration (24 h access to water following 48 h of water deprivation) also elevated kidney cul-5 mRNA levels. Water deprivation did not significantly alter cul-5 mRNA levels in the brainstem, cerebellum, hippocampus, lung or liver. Since cul-5 appears to be linked to proteosome-mediated protein degradation, it may have a role in protein regulation under conditions of osmotic stress. PMID- 12631467 TI - Suppression of oxidative neuronal damage after transient middle cerebral artery occlusion in mice lacking interleukin-1. AB - Interleukin-1 (IL-1) contributes to ischemic neurodegeneration. However, the mechanisms regulating action of IL-1 are still poorly understood. In order to clear this central issue, mice that were gene deficient in IL-1alpha and beta (IL 1 KO) and wild-type mice were subjected to 1-h transient middle cerebral artery occlusion (tMCAO). Expression levels of IL-1beta and IL-1 receptor I (IL-1RI) were then examined. Generation of peroxynitrite and the expression of mRNAs for nitric oxide synthase (NOS) subtypes were also determined. Immunostaining for IL 1beta was increased from 6 h and peaked at 24 h after tMCAO in the microglia and macrophage. The immunoreactivities of IL-1RI were increased progressively in the microvasculature and neuron-like cells of the ipsilateral hemisphere. Infarct volumes were significantly lower in IL-1 KO mice compared with wild-type mice 48 h after tMCAO (P<0.01). The immunoreactivities of 3-nitro-L-tyrosine were determined in the neurons and microvasculature 24 h after tMCAO and were significantly decreased in the IL-1 KO mice compared to wild-type mice. In addition, expression levels of NOS mRNA in IL-1 KO mice were lower than that measured in wild-type mice. These results indicate that IL-1 is up-regulated and may play a role in neurodegeneration by peroxynitrite production during ischemia. PMID- 12631468 TI - Microglia in the olfactory bulb of rats during postnatal development and olfactory nerve injury with zinc sulfate: a lectin labeling and ultrastrucutural study. AB - Using isolectin (GSA I-B4) as a marker, this study examined the possible alterations of lectin-labeled membranous glycoproteins in microglial cells in the olfactory bulb of normal development and under experimentally induced degeneration. In light microscopy, several morphological types of microglial cells representing different degrees of cell differentiation were distributed in the bulb laminae. A gradient of microglial differentiation extending from the intermediate to superficial and intermediate to deep occurs in the bulb layers. The differentiation gradient and lectin labeling pattern of microglial cells in the developing bulb resembled those in other areas of the brain tissues. Differentiating microglia showed a gradual diminution of lectin staining when the nascent round cells transformed into the mature ramified cells. Microglia in the external plexiform layer of the olfactory bulb were the first to mature and the cells expressed very weak lectin reactivity. In mature or adult rats, some microglial cells showing intense lectin labeling were observed in the olfactory nerve layer, granule cell layer and subependymal layer. Ultrastructurally, lectin labeling was localized at the trans saccules of the Golgi apparatus. Microglial cells in other bulb laminae, however, exhibited a negative reaction for the isolectin at the Golgi apparatus. Following intranasal irrigation of zinc sulfate, some microglial cells in the olfactory nerve layer and glomerular layer were activated to become phagocytic cells with increased lectin labeling at their ramified processes. GSA I-B4 staining was also localized at their trans saccules of the Golgi apparatus. The lectin labeling pattern of these phagocytic cells resembled that of differentiating microglia in postnatal bulbs, suggesting that bulb microglia in the lesioned sites were activated through cell dedifferentiation into macrophages. PMID- 12631469 TI - DOPA cyclohexyl ester potently inhibits aglycemia-induced release of glutamate in rat striatal slices. AB - Brain ischemic insult causes glutamate release and resultant neuronal cell death. We here show that L-3,4-dihydroxyphenylalanine (DOPA) is a positive regulatory factor for glutamate release elicited by a mild brain insult using in vitro superfused rat striatal slices as a model system. Glucose deprivation for 18 min elicited release of glutamate, DOPA and dopamine (DA). Either tetrodotoxin (TTX) (1 microM) or alpha-methyl-p-tyrosine (alpha-MPT) (1 mM), a tyrosine hydroxylase inhibitor reduced markedly each of these releases. NSD-1015 (20 microM), an aromatic L-amino acid decarboxylase inhibitor restored the inhibition by alpha MPT of glutamate and DOPA but not DA release. DOPA cyclohexyl ester (DOPA CHE) (0.3-1 microM), a competitive DOPA antagonist, concentration-dependently suppressed aglycemia-induced glutamate release, the effect which was mimicked neither by S-sulpiride nor SCH23390, a DA D(1) or D(2) receptor antagonist, respectively. Zonisamide (1-1000 microM), an anticonvulsant or YM872 (1 microM), an alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) a receptor antagonist produced no effect on aglycemia-induced glutamate release. DOPA CHE thus showed a relatively potent inhibitory action on aglycemia-induced glutamate release among several neuroprotective agents tested. PMID- 12631470 TI - Low dose effects of bisphenol A on sexual differentiation of the brain and behavior in rats. AB - There is an endocrinological concern that environmental endocrine disrupters (EEDs) may influence sexual differentiation. Bisphenol A (BPA), one of EEDs, is released from polycarbonate plastics, and has been detected in the human umbilical cord. In this study, we examined the effect of BPA on the sexual differentiation of open-field behavior and the sexually dimorphic nuclei in the brain in the offspring of rats exposed to BPA during the fetal and suckling periods at a dosage below the human tolerable daily intake (TDI) level. In the control group, females were more active in the open field and had a larger locus coeruleus (LC) volume than males. BPA abolished and inverted the sex differences of the open-field behavior and the LC volume, respectively, without affecting the reproductive system. We also compared the effects of estrogenic compounds, diethylstilbestrol (DES) and resveratrol (RVT), to that of BPA because of their structural similarities. DES affected the open-field behavior, LC volume and reproductive system, while RVT affected the LC volume and the reproductive system. These results suggest that the brain is highly sensitive to BPA at a dosage below TDI and that the disrupting effects of BPA on sexual differentiation may vary from those of RVT and DES. PMID- 12631472 TI - Construction and evaluation of a eukaryotic expression plasmid for stable delivery using attenuated Salmonella. AB - An approach to enhancing the stability of eukaryotic expression plasmids for delivery using attenuated Salmonella has been evaluated. The expression apparatus and beta-galactosidase gene from the expression plasmid, pCMVbeta, was cloned into the low copy number plasmid pLG339. The resulting construct, pLGbetaGAL, was shown to have a lower copy number than pCMVbeta in Salmonella enterica var Typhimurium aroA strain SL7207. Furthermore, beta-galactosidase-specific antibody was induced in mice following intramuscular inoculation with pLGbetaGAL as naked DNA. Following oral administration of mice with SL7207/pCMVbeta, recombinants could not be detected in tissues 3 days after inoculation. In comparison, SL7207/pLGbetaGAL recombinant bacteria could be detected in the Peyer's patches and spleens indicating that the Salmonella strain was stable. However, both SL7207/pCMVbeta and SL7207/pLGbetaGAL failed to induce beta-galactosidase specific IgG in vivo. The mechanism by which attenuated Salmonella are able to release heterologous DNA for antigen processing and presentation is not yet understood. These results suggest that the mechanism needs to be further elucidated in order to rationally improve the system. PMID- 12631471 TI - Functionally essential neuronal population of the facial motor nucleus. AB - Cranial nerve impairment is one of the more serious complications in neurosurgery. Nevertheless, the important question of how many neurons are required for cranial nerve functions remains unanswered. The VIIth cranial nerve (facial nerve) in mice was subjected to graded crush injuries. After assessment of the facial function, the number of uninjured, healthy facial motor neurons was quantified with a retrograde neuronal tracer. We report that normal facial function is preserved if intact neurons account for more than 56% of the control value, while complete facial paralysis occurs if intact neurons are reduced to less than 32% of the control value. PMID- 12631473 TI - Involvement of lipooligosaccharides of Haemophilus influenzae and Neisseria meningitidis in defensin-enhanced bacterial adherence to epithelial cells. AB - Stimulated neutrophils release a variety of antimicrobial peptides, including neutrophil defensins (HNP1-4). We have previously reported that neutrophil defensins enhanced the adherence of Haemophilus influenzae and Neisseria meningitidis to cultured respiratory epithelial cells. In this study, the effect of defensins on the adherence of H. influenzae and N. meningitidis lipooligosaccharide (LOS) mutants to epithelial cells was tested. Neutrophil defensins enhanced the adherence of the oligosaccharide mutants of H. influenzae and N. meningitidis, whilst the adherence of the lipid A mutants B29 of H. influenzae and lpxL1 and lpxL2 of N. meningitidis was not or only moderately stimulated by neutrophil defensins. The adherence of the N. meningitidis LOS negative mutant lpxA was not enhanced by defensins. These findings suggested that the secondary fatty acids of lipid A were involved in the defensin-enhanced adherence. LOS from strain H44/76 or HNP-LOS complexes did not affect or stimulate the adherence of N. meningitidis, although the defensin-enhanced adherence is specific for certain bacterial species having LOS in their outer membrane. These results indicated that LOS is involved in the defensin-enhanced adherence. However, the mechanism by which defensins and LOS interact with epithelial cells to promote bacterial adherence remains to be resolved. PMID- 12631474 TI - Genetic manipulation of immunoglobulin binding proteins of Haemophilus somnus. AB - The relationship of the 76kDa immunoglobulin binding, surface antigen (p76) of Haemophilus somnus to the high molecular weight immunoglobulin binding proteins (HMW IgBPs) was investigated. The kanamycin resistance gene from pLS88 was used via homologous recombination with allelic exchange to replace a portion of the gene encoding IgBPs of H. somnus strain 8025. Recombinants were shown by Western immunoblotting to express and secrete truncated antigens of approximately 200kDa and not to produce p76. The truncated HMW IgBP variants retained the ability to bind bovine IgG2 by the Fc portion as demonstrated by Western immunoblotting against IgG2 anti-DNP. This data indicated that the deleted 1.8kb BglII fragment was not required for secretion or immunoglobulin Fc binding by the HMW IgBPs but was required for expression of the downstream p76 gene. Functional studies showed that, in addition to Fc binding of IgG2 to truncated HMW IgBPs, the mutant strain 8025 Kan1 was equally resistant to killing by mouse complement but less virulent than the wild type parent (8025) in a mouse septicemia model of H. somnus infection. However, mutant strain 8025 Kan1 did adhere less well than the wild type to bovine pulmonary artery endothelial cells. It is probable that p76 and the missing peptides of the HMW IgBPs play a role in this aspect of virulence and perhaps other aspects. PMID- 12631475 TI - Adherence of Mycoplasma bovis to bovine bronchial epithelial cells. AB - Mycoplasma bovis is responsible for considerable economic losses in cattle due to pneumonia, arthritis and mastitis. As the agent was shown to be capable of adhering to neutrophils and embryonic bovine lung (EBL) cells and invading the respiratory epithelium it is highly desirable to improve our understanding of cytadherence processes. Although several surface proteins likely to be directly involved in this initial stage of interaction between pathogen and host cells have been identified, these findings mainly referred to type strain PG45 adhering to the continuous EBL cell line. The present study provides new and complementary data about cytadherence of M. bovis based on adherence of various radiolabeled strains to a primary culture of bovine bronchial epithelial (BBE) cells using a standardized adherence assay. M. bovis was shown to adhere specifically to the primary culture of BBE cells. Inhibition of adherence was observed upon addition of monoclonal antibodies (MAbs), trypsin treatment of mycoplasmas, and competition with non-radiolabeled mycoplasma cells. Interestingly, three MAbs against proteins involved in adherence to EBL cells failed to inhibit significantly the adherence to BBE cells. On the other hand, significant reduction of adherence rates by MAbs 2A8 and 9F1 directed against epitopes of variable surface lipoproteins VspC and VspF, respectively, demonstrated the involvement of these proteins in adherence of M. bovis to primary culture of BBE cells. PMID- 12631476 TI - Systemic and enteric colonization of pigs by a hilA signature-tagged mutant of Salmonella choleraesuis. AB - Although host adapted to pigs, Salmonella enterica serovar Choleraesuis (S. choleraesuis) can induce a virulent foodborne salmonellosis in humans. To directly investigate virulence factors of S. choleraesuis, we extended the functional genomics approach of signature-tagged mutagenesis to S. choleraesuis and pigs. When a test pool of 45 randomly signature-tagged null mutants was inoculated orally and intraperitoneally in pigs, one of the mutants that failed to colonize by either route was tagged in hilA. In the broad host range serovar S. typhimurium, hilA regulates invasion genes in the first pathogenicity island and is required for enteric but not systemic infections in mice experimentally infected with S. typhimurium. The pool of tagged S. choleraesuis null mutants was a complex mix inoculated in pigs. When pigs were challenged with an equal mixture of the hilA mutant and wild type bacteria, the hilA mutant was at a competitive disadvantaged (attenuated) in pigs inoculated orally but not in intraperitoneally inoculated pigs. Our data support that hilA in S. choleraesuis infections of pigs has a role in enteric but not systemic infections similar to that of S. typhimurium in the murine model of human typhoid fever. The role of hilA may be conserved across Salmonella serovars and host species. PMID- 12631477 TI - Mutations in hns reduce the adherence of Shiga toxin-producing E. coli 091:H21 strain B2F1 to human colonic epithelial cells and increase the production of hemolysin. AB - Shiga toxin-producing Escherichia coli (STEC) 091:H21 strain B2F1, an isolate from a patient with the hemolytic uremic syndrome (HUS), produces elastase activatable Shiga toxin (Stx) type 2d and adheres well to human colonic epithelial T84 cells. This adherence phenotype occurs even though B2F1 does not contain the locus of enterocyte effacement (LEE) that encodes the primary adhesin for E. coli O157:H7. To attempt to identify genes involved in binding of B2F1 to T84 cells a bank of mini-Tn5phoACm(r) transposon mutants of this strain was generated. Several of these mutants exhibited a reduced adherence phenotype, but none of the insertions in these mutants were within putative adhesin genes. Rather, insertional mutations within hns resulted in the loss of adherence. Moreover, the hns mutant also displayed an increase in the production of hemolysin and alkaline phosphatase and a loss of motility with no change in Stx2d activatable expression levels. When B2F1 was cured of the large plasmid that encodes the hemolysin, the resulting strain adhered well to T84 cells. However, an hns mutant of the plasmid-cured B2F1 strain exhibited a reduction in adherence to T84 cells. Taken together, these results indicate that H-NS regulates the expression of several genes and some potential virulence factors in the intimin negative B2F1 STEC strain and that the large plasmid is not required for T84 cell colonization. PMID- 12631478 TI - Fingers, hands or patients? The concept of independent observations. AB - In hand surgery trials, it is often possible to take several measurements from the same patient, because many disorders here affect bilateral or multiple structures, such as the hand itself, the finger joints or the tendons. Most conventional statistical analyses that take place on the level of hands, digit rays or joints rather than patients violate the assumption that observations should be independent. Furthermore, ignoring the multiplicity of data inflates sample size and thus may lead to spurious significance. This article describes three options to deal with such problems. First, the analysis can simply be restricted to only one measurement per patient. Second, a self-controlled design may be advantageous for conditions that usually have a bilateral pattern. Third, complex statistical modelling (involving generalized estimating equations) can be used to analyse all available measurements with adjustment for data dependency. PMID- 12631479 TI - What determines the costs of repair and rehabilitation of flexor tendon injuries in zone II? A multiple regression analysis of data from southern Sweden. AB - The epidemiology and costs of repair and rehabilitation of zone II flexor tendon injuries in 135 patients from the southern part of Sweden were analysed. The little finger was most frequently injured (43%), usually with a knife (46%), and 30% of the injuries were work related. Total median costs within the health-care sector for the injuries were SEK 48,500 (1 EURO=9.23 SEK, 4/1/2002). Costs in other sectors were SEK 93,000. Active mobilization or mobilization with rubber band traction increased costs within the health-care sector (SEK 7400 or SEK 6000, respectively) but improved range of movement (5-7%). Immobilization had a higher complication rate (rupture or need for secondary procedures), which in itself increased total costs by 57%. Non-linear effects were found between age and costs within the health-care sector and the outcome. PMID- 12631481 TI - Flexor tendon graft attachment: a review of methods and a newly modified tendon graft attachment. AB - A review of the described methods of attachment of flexor tendon grafts to the distal phalanx is presented. The authors advocate the previously described method of attachment consisting of passage of the tendon graft through the pulp with anchorage to the nail. A new modification of this technique is presented, facilitating accurate dissection and placement of the graft with minimal trauma. PMID- 12631480 TI - Efficacy of ADCON-T/N after primary flexor tendon repair in Zone II: a controlled clinical trial. AB - A prospective double-blind, randomized, controlled clinical trial was conducted to assess the use of ADCON-T/N after flexor tendon repair in Zone II. Forty-five patients with 82 flexor tendon repairs in 50 digits completed the study. ADCON T/N was injected into the tendon sheath after tenorrhaphy in the experimental group while the control group was not treated with ADCON-T/N. ADCON-T/N had no statistically significant effect on total active motion at 3, 6 and 12 months but the time taken to achieve the final range of motion was significantly shorter in treated patients. ADCON-treated patients had a higher rupture rate but this was not significant. PMID- 12631482 TI - The dorsal flap of the first web. AB - In the first web space region, there are communications between the dorsal arteries which arise from the dorsal branch of the radial artery and the digital arteries of the thumb and the index finger. These allow a distally based flap to be raised in the first dorsal intermetacarpal area. This flap has been used in 15 cases of soft-tissue loss from the thumb and index finger. The donor sites were closed primarily in all but two patients. There were no complications, and the results show that this flap is useful for soft-tissue defects on the tip and the palmar and dorsal surfaces of the thumb. Moreover, it may be used as a "cross finger" flap. PMID- 12631483 TI - Long-term results of syndactyly correction: full-thickness versus split-thickness skin grafts. AB - In order to compare the long-term results of full-thickness and split-thickness skin grafts after the correction of congenital syndactyly, 27 patients have been investigated after an average follow-up of 21 years. Post-operative functional and cosmetic results have been assessed by patient records, questionnaires and physical examination. The webs that had received split-thickness grafts showed more flexion and extension lags and the overall spreading of the operated fingers was significantly decreased compared to the control fingers. On the other hand, more re-operations because of web creep had to be performed after full-thickness grafts. Hyperpigmentation and hair growth in the grafts was found in most of the full-thickness grafts, while breakdown of the graft was found in some of the split-thickness grafts. Therefore, based on the results of this study, either full- or split-thickness skin grafts can be used when treating of congenital syndactyly. PMID- 12631484 TI - Vascular anatomy of the palmar surfaces of the distal radius and ulna: its relevance to pedicled bone grafts at the distal palmar forearm. AB - The potential for harvesting vascularized bone grafts from the palmar surface of the distal radius has been studied in 40 arms of fresh cadavers which had previously been injected with coloured latex solution. It was found that vascularized grafts can be pedicled on the radial part of the palmar carpal arterial arch. If a longer pedicle is required, the bone graft can be pedicled on the anterior branch of the anterior interosseous artery with retrograde flow occurring from the palmar carpal arch. PMID- 12631485 TI - Dynamic external fixator for pilon fractures of the proximal interphalangeal joints: a simple fixator for a complex fracture. AB - We describe a very cheap, simple and effective dynamic external fixator for treatment of pilon fractures of the proximal interphalangeal joint. At final follow-up, nine such fractures had regained an average range of motion of 79 degrees (range, 65-90 degrees ). There was high patient satisfaction and there were no serious complications. PMID- 12631486 TI - Closed reduction transarticular Kirschner wire fixation versus open reduction internal fixation in the treatment of Bennett's fracture dislocation. AB - Thirty two patients with fracture dislocations of the base of the thumb metacarpal with a single large fracture fragment (Bennett's fracture) were either treated by open reduction and internal fixation or closed reduction and percutaneous transarticular Kirschner wiring. All were assessed at a mean follow up of 7 (range 3-18) years. Patients with an articular step off more than 1mm were excluded. The type of treatment did not influence the clinical outcome or the prevalence of radiological post-traumatic arthritis. The percutaneous group had a significantly higher incidence of adduction deformity of the first metacarpal. This was attributed to Kirschner wire placement near the fracture line or in the compression zone of the fracture, resulting in loss of reduction. This however did not result in an inferior outcome. PMID- 12631487 TI - Neurophysiological prediction of outcome in obstetric lesions of the brachial plexus. AB - Seventy-three children with slow recovery after obstetric lesion of the brachial plexus (biceps function returning after 3 months of age) and with relatively favourable neurophysiological investigation were followed until a mean age of 4.3 years. Predictions for C6 and C7 were confirmed in 92% and 96% of the cases, respectively. Predictions for C5 were confirmed in a smaller proportion of cases (78%). The inability to record nerve action potentials for C5, and the high frequency of secondary shoulder pathology are reasons for this. In all, 33 of the 73 children came to operation for medial rotation contracture (11) or posterior subluxation/dislocation (21); there was one case operated for inferior contracture. PMID- 12631488 TI - Tendon transfer to reconstruct wrist extension in children with obstetric brachial plexus palsy. AB - This study reports on 20 children with obstetric brachial plexus palsy who underwent a tendon transfer to reconstruct wrist extension. The mean age at the time of tendon transfer was 8 years. There were seven patients with Erb's palsy and the remaining 13 had total palsy. The flexor carpi ulnaris was utilized 15 times and the flexor carpi radialis five times. The transferred tendon was sutured to the tendon of the extensor carpi radialis brevis. The result of the transfer was assessed according to a modified Medical Research Council (MRC) muscle grading system. A good result was obtained in 18 patients (modified MRC grade of 4) and a fair result (modified MRC grade of 3) in two. The choice of tendon transfer to reconstruct the wrist drop deformity in various conditions including adult traumatic brachial plexus injuries is discussed. PMID- 12631489 TI - Relationship between joint laxity and radio-ulno-carpal joint morphology. AB - The relationships between wrist laxity, ulnar variance, sigmoid notch inclination, and lunotriquetral motion were analysed in 60 normal volunteers. A strong correlation between ulnar length and sigmoid notch inclination was found for the entire group. Joint laxity was found to correlate with ulnar variance and lunotriquetral mobility in women, but not in men. The greater the laxity, the shorter the ulna and the greater the lunotriquetral motion during radial to ulnar deviation. These results support the concept that laxity increases the vulnerability of the wrist to injury. PMID- 12631490 TI - Palmar cortical angle of the distal radius: a radiological study. AB - Fifty lateral radiographs of normal wrists were viewed to determine the palmar cortical angle of the distal radius. The palmar cortical angle is different to its previously described palmar tilt or angulation. The mean value for the palmar cortical angle was 37 degrees (range, 26-50 degrees). This may be clinically important in the design of palmar plates for the distal radius. PMID- 12631491 TI - Hand function tests and questions on hand symptoms as related to the Stockholm workshop scales for diagnosis of hand-arm vibration syndrome. AB - The severity of hand-arm vibration syndrome (HAVS) is usually graded according to the Stockholm workshop scales. Although the Stockholm workshop scales are regarded the gold standard for assessing the severity of HAVS, they are based primarily on subjective symptoms. The aim of the present study was to explore the agreement between Stockholm workshop scales and the outcome from ten well-defined clinical tests commonly used in hand rehabilitation for assessment of hand function. One hundred and eleven vibration-exposed workers participated in the study. Ten objective tests of hand function and four questions on subjective hand symptoms were included. The results indicated that, out of these tests, perception of vibration, perception of touch/pressure and dexterity showed a moderate agreement with Stockholm workshop scales. Among specific questions on hand symptoms, cold intolerance and pain showed a high agreement with Stockholm workshop scales. It is concluded that defined objective tests combined with directed questions on specific hand symptoms, together with the Stockholm workshop scales, may be helpful for diagnosing HAVS. PMID- 12631492 TI - Palmar wrist ganglion: does intervention improve outcome? A prospective study of the natural history and patient-reported treatment outcomes. AB - A prospective cohort study was undertaken to observe the long-term outcome of different treatments for palmar wrist ganglia. One hundred and eighty-two patients agreed to participate in the study. One hundred and fifty-five patients (88%) responded at 2 or 5 years. Seventy-nine had been treated by surgical excision, 39 by aspiration and 38 by reassurance alone. At 5 years no significant differences were observed in the recurrence rates which were 42% after excision of a palmar wrist ganglion and 47% (19 of 39) after aspiration. Twenty of the 39 untreated ganglia had disappeared spontaneously. Eighty-five per cent of the patients were satisfied irrespective of treatment. Patients having surgery had a complication rate of 20% and took more time off work (14 days). Significantly more patients in the untreated group felt the persistent ganglion was unsightly. The patient evaluation measure scores were similar. At 2 and 5 year follow-up, regardless of treatment, no difference in symptoms was found, regardless of whether the palmar wrist ganglion was excised, aspirated or left alone. One in four wrists remained weak regardless of treatment or disappearance of the ganglion. PMID- 12631493 TI - Ganglion-associated ulnar tunnel syndrome treated by ultrasonographically assisted aspiration and splinting. AB - We describe a case of ulnar nerve compression at the wrist due to a ganglion. This was treated by aspiration of the ganglion under ultrasonography and splinting because the patient was pregnant. The ulnar nerve palsy resolved completely and the ganglion disappeared. A follow-up ultrasonographic examination after 2 years showed no recurrence of the ganglion. PMID- 12631494 TI - Italian version of the Disability of the Arm, Shoulder and Hand (DASH) questionnaire. Cross-cultural adaptation and validation. AB - An Italian version of the Disability of the Arm, Shoulder and Hand (DASH) questionnaire has been devised and its reliability and validity have been assessed in a cross-sectional study of 108 consecutive patients with upper extremity pathologies. A sub-sample of 30 patients was used to assess re-test reliability. The principal DASH scale showed a high correlation with other patient-oriented measures and demonstrated good reproducibility, consistency and validity, which were similar to those for other languages' versions of DASH. These findings suggest that the evaluation capacities of the Italian DASH are equivalent to those of other language versions of the DASH. PMID- 12631495 TI - Lumbrical tears in rock climbers. AB - Performance rock climbing places high demands on the hand and may lead to specific injuries. In a "one-finger-pocket" hold, the interphalangeal joints remain in 20-40 degrees flexion. To increase the maximum force of the holding finger by the quadriga effect, the interphalangeal joints of the adjacent fingers become almost maximally flexed. Holding a "one-finger-pocket" with the ring or small finger leads to a shift of the deep flexor tendons which increases the distance between the two adjacent origins of either the third or the fourth lumbrical. This may cause disruption and tear of that muscle. An organized haematoma in the third lumbrical was visible by ultrasonography in one of the three cases described. PMID- 12631496 TI - A simple and fail safe method for digital tourniquet. PMID- 12631498 TI - Re: Silicone arthroplasty for trapeziometacarpal arthritis. PMID- 12631499 TI - The role of intra-oral manipulation in the perception of sensory attributes. AB - To gain insight into the effect of oral processes on perception, we defined a set of five specific oral manipulations and investigated their effects on the perception of low and high fat versions of two semi-solid foodstuffs, vanilla custard desserts and mayonnaises. Behavior modifications ranged from simply placing the stimulus on the tip of the tongue to vigorously moving it around in the mouth. Sensory ratings for mouth-feel and flavor attributes were made 5s after placing the stimulus in the mouth, and after-feel attributes were rated immediately after swallowing. Most attributes showed a similar pattern, with lowest attribute ratings where the tongue's movement was restricted and gradually increasing ratings with increasing complexity of the tongue movements. An individual's normal oral processing behavior typically resulted in the most intense sensations of flavor and mouth-feel. Residence time for all mouth-feel attributes, except prickling, was determined by the time required for tongue movements. The exact tongue movements required for sensations appeared to be related to food groups and individual foods, rather than to specific mouth-feel attributes. PMID- 12631500 TI - The perceived healthiness of functional foods. A conjoint study of Danish, Finnish and American consumers' perception of functional foods. AB - Functional foods presumably enable the consumer to lead a healthier life without changing eating habits. Whether consumers accept this proposition or not is potentially influenced by their perceptions of the healthiness of the processing methods, enrichment components, food-types, and health claims used in the production and marketing of functional foods. Because consumers may perceive functional enrichment as interfering with nature, cultural values pertaining to man's manipulation of nature may also influence consumer acceptance of functional foods. The purpose of the study described here is to clarify to which extent Danish, Finnish and American consumers' perceptions of the healthiness of functional foods are explained by the factors mentioned above. The general results indicate that values pertaining to man's manipulation of nature is only modestly related to the acceptance of functional foods, whereas the use of different health claims, processing methods, enrichments, product types, and especially the interactions between the two latter, are important determinants of consumers' perceptions of the healthiness of functional foods. PMID- 12631501 TI - The relative-reinforcing value of food under differing levels of food deprivation and restriction. AB - Food deprivation and restriction both increase food consumption. Food deprivation also increases the reinforcing value of food, but it is unknown if food restriction alone or combined with deprivation increases the reinforcing value of food. Forty, normal-weight, college-aged, unrestrained females were randomized to one of four conditions that crossed food deprivation (Dep) and restriction (Res): Dep/Res, Dep/No Res, No Dep/Res, No Dep/No Res. All participants arrived at least 13h food-deprived, and non-deprived participants consumed at least 365cal from a drink during the session. Restriction was manipulated by placing snack food in front of participants, without access for 15min during the session, or having no snack food placed in front of participants. Following the experimental manipulations, participants completed a computer choice task to determine the reinforcing value of food. Repeated measures analysis of variance found a significant main effect of deprivation (p<0.05) and trials (p<0.001) for food points earned. Deprived participants found food to be more reinforcing, and the reinforcing value of food decreased over time. This suggests that while short term food deprivation increases the relative-reinforcing value of food, short term food restriction has no effect on the relative-reinforcing value of food in unrestrained eaters. PMID- 12631502 TI - Food choices among newly married couples: convergence, conflict, individualism, and projects. AB - Individuals entering marriage merge their personal food systems to create a joint spousal food system. This investigation examined newly married couples' food choices. Qualitative interviews conducted with 20 couples as they entered marriage and again 1 year later revealed that some reported initial food homogamy, while others had varying degrees of compatibility in food preferences and tolerances. All couples reported some dietary convergence. Convergence was initially symmetrical (similar merging for both partners) or asymmetrical (one partner changed more), and sometimes convergence patterns evolved further. Unresolved food choice negotiations led to food conflicts. Prior food broadening experiences, initial dietary congruity, health/weight philosophies, foodwork, and commensal patterns were conditions shaping couples' food choice negotiations and dietary convergence patterns. Some food individualism persisted for most partners, helping to alleviate food choice conflicts. Food projects occurred when one partner was committed to molding the other's food choices. Overall, food choice negotiations involving both convergence and conflict were important components of marital adjustments that may have later health consequences. PMID- 12631503 TI - Effect of moderate intensity exercise on acute energy intake in normal and overweight females. AB - The purpose of this study is to determine the impact of moderate intensity exercise on acute post-exercise energy intake (EI) in normal and overweight females. Twelve normal weight (BMI<25), and 12 over weight (BMI>25) inactive women (mean age 35+/-8) participated in a 2-day experimental study. On the exercise day, participants were asked to walk on a treadmill at 60% maximum heart rate for 1h. After the exercise, participants selected and ate food at a university cafeteria. Post-exercise EI was compared to a non-exercise day. A single mixed model repeated 2 x 2 ANOVA was used to determine the impact of exercise on post-exercise EI. The overweight women consumed significantly more (p<0.02) than the normal weight women on both experimental days and there was a significant (p<0.03) difference in fat intake. The higher post -exercise EI of the overweight group may have been due to the energy needs associated with a higher BMI but it is also possible that cognitive factors, food availability and environment may influence post-exercise EI. PMID- 12631504 TI - Relationship of gender and eating disorder symptoms to reported cravings for food: construct validation of state and trait craving questionnaires in Spanish. AB - Using confirmatory factor analysis, we cross-validated the factor structures of the Spanish versions of the State and Trait Food Cravings Questionnaires (FCQ-S and FCQ-T; ) in a sample of 304 Spanish college students. Controlling for eating disorder symptoms and food deprivation, scores on the FCQ-T were higher for women than for men, but no sex differences were observed on the FCQ-S. Eating disorder symptomatology was predictive of trait cravings, whereas food deprivation was predictive state cravings. Trait cravings, but not state cravings, were more strongly associated to symptoms of anorexia and bulimia nervosa than with other psychopathology. We suggest that cravings can be conceptualized as multidimensional motivational states and that our data support the hypothesis that food cravings are strongly associated with symptoms of bulimia nervosa. PMID- 12631505 TI - Feeding behaviour in young children who fail to thrive. AB - One-year-old children who failed to thrive in infancy were identified through a specialist clinical service using a conditional weight gain criterion which identified the slowest gaining 5%. Control children of the same age and sex were recruited from the same local geographical area and had the same primary care physician. The food intake and feeding behaviour of the groups was compared using a detailed observational micro-analysis of a lunchtime meal, using a behavioural coding scheme developed for use over the weaning period. Both food and fluid intake at the test meal were significantly lower in the children who failed to thrive than the controls. There was no significant difference in the energy density of the foods they consumed. As recorded in the behaviour counts at the meal, the mothers of the children who failed to thrive fed them as much as or more than the control mothers fed their children. The children who failed to thrive tended to refuse or reject the offered food more, and also fed themselves significantly less often than the controls. These behavioural differences during the meal accounted for about one third of the difference in energy intake between the groups. PMID- 12631506 TI - Perceptions and reasons for choice of fat- and fibre-containing foods by Swedish schoolchildren. AB - This study investigated schoolchildren's breakfast choices focusing on their reported usual intake and perceptions of fat-reduced or fibre-enriched alternatives. A total of 184 eleven- to fifteen-year-old were asked to participate, and 181 completed the study. Using pictures of food items, the children were instructed to reproduce their usual breakfast, and then breakfasts that would be: healthy, tasty, consumed by their parents, or approved by their parents. Participants' usual breakfast included breads and breakfast cereals containing a higher fibre content than that of the breakfast reported as most palatable. Their usual breakfast also contained products with less fibre than was the case for their healthy or perceived parental norm breakfasts. This may be interpreted as evidence that high-fibre bread and breakfast cereals are perceived as healthy but that consumption of these products is limited by palatability. In contrast to the ambivalence towards cereal products, corresponding disagreements between breakfasts were not observed for milk, and only a small systematic disagreement was observed for spread. Thus, the majority consumed the kind of milk and spread perceived as being healthy, tasty and parentally sanctioned despite the fact that full-fat products were frequently consumed. In conclusion, the results suggest that taste may be a barrier to consumption of high-fibre products, while consumption of low-fat products appears to be impeded by health beliefs. PMID- 12631507 TI - Perceptual channels for the texture of a food. AB - The present study makes a start on characterising the cognitive processes by which physical effects of eating on the senses are transformed into quantitative judgments about perceived characteristics of a food. It was hypothesised that there is a discrete perceptual channel for an aspect of texture sensed during each of the three initial movements of eating a piece of a cookie. The results showed that the force required for initial compression of the surface of the biscuit related to how 'crisp' it was. Scores on 'hard' were sensitive to higher forces being required to bite off a piece. Ratings of crunchiness responded to both amplitude and frequency of the cracks opened up in this heterogeneously structured material during the first crushing of the bitten piece. These findings are being pursued to identify the stimulation patterns more precisely and to measure how the percepts are integrated into judgments of overall texture. PMID- 12631508 TI - Correlations in perceived food use between the family food preparer and their spouses and children. AB - Previous research has demonstrated that family members' eating habits are similar. We hypothesized that family members' eating habits would be predicted by the eating habits of the person who prepares the majority of the family's meals and the number of meals the family shares. Participants were 282 members of religious organizations who identified themselves as family food preparers (FFPs), and provided information about their own and their family members' eating habits. Results revealed that FFP fruit and vegetable intake predicted the fruit and vegetable intake of spouses, children, and adolescents (p<0.01), and that FFP consumption of high-fat foods predicted the consumption of high-fat foods of spouses and children (p<0.01). Child fruit and vegetable consumption was also influenced by shared meals: the more meals the child shared with the FFP, the stronger the relationship of FFP fruit and vegetable intake with child fruit and vegetable intake (p<0.05). These findings indicate that dietary interventions targeting the FFP may benefit other family members. PMID- 12631509 TI - Impaired visual discrimination learning in anorexia nervosa. AB - The primate dopamine system is involved in appetitively motivated behaviours, including certain forms of learning, for example, visual discrimination learning. Furthermore, food restriction in animals and anorexia in humans is associated with impaired dopamine signaling. Based on this, we hypothesized that patients with anorexia nervosa (AN) would show a deficit in visual discrimination learning. In a dynamic categorization task involving the learning of a series of two-alternative forced-choice visual discriminations, conceptually identical to one shown to activate dopamine neurons in primates, and sensitive to dopaminergic manipulations in humans, patients with AN showed a deficit in learning that was most pronounced in the early stages of acquisition. In contrast, AN showed spared performance on a pattern recognition memory test sensitive to medial temporal lobe lesions, but insensitive to dopaminergic manipulations. We conclude that impaired appetitive function in patients with AN extends to include deficits in visual discrimination learning, and that this deficit represents indirect evidence for altered dopaminergic neurotransmission in AN. PMID- 12631510 TI - Generalized 3D nonlinear transformations for medical imaging: an object-oriented implementation in VTK. AB - We have contributed an efficient, object-oriented implementation of 3D nonlinear transformations to the Visualization Toolkit that can be applied to a wide variety of data types, including images and polygonal meshes. The transformations are performed via thin-plate splines or via interpolation of a regular lattice of displacement vectors, and are part of a framework that is easily extensible to other nonlinear transformation types. In this paper we demonstrate application of these transformations in medical imaging in general and image-guided surgery in particular, and present a series of performance benchmarks. PMID- 12631511 TI - A comparative study of warping and rigid body registration for the prostate and pelvic MR volumes. AB - A three-dimensional warping registration algorithm was created and compared to rigid body registration of magnetic resonance (MR) pelvic volumes including the prostate. The rigid body registration method combines the advantages of mutual information (MI) and correlation coefficient at different resolutions. Warping registration is based upon independent optimization of many interactively placed control points (CP's) using MI and a thin plate spline transformation. More than 100 registration experiments with 17 MR volume pairs determined the quality of registration under conditions simulating potential interventional MRI-guided treatments of prostate cancer. For image pairs that stress rigid body registration (e.g. supine, the diagnostic position, and legs raised, the treatment position), both visual and numerical evaluation methods showed that warping consistently worked better than rigid body. Experiments showed that approximately 180 strategically placed CP's were sufficiently expressive to capture important features of the deformation. PMID- 12631512 TI - 3D time of flight MR angiography: acquisition with small field of view and low phase encodes. AB - The aim of the study was to evaluate and compare the image quality of the 3D TOF MRA acquired with a small FOV and low phase encodes with those MR angiographic images acquired with standard pulse sequence parameters. Twenty patients who were referred to our institution for MR imaging of the brain and strictly satisfied the selection criteria were included in this study. Apart from the routine protocol for MR imaging of the brain, 3D TOF MRA of the circle of Willis with a small FOV and a standard FOV were performed. The image quality of all MRA was evaluated by two independent observers who were blind to the pulse sequence parameters. From the standard FOV MRA, 22.5, 12.5, and 5% of the patients were graded as mild, moderate, and severe stenosis of the internal carotid artery, respectively. On the contrary, no apparent stenosis was observed from the small FOV MRA with low phase encodes. Regarding the reduction in MR artifacts and acquisition time achieved with the small FOV 3D TOF MRA with low phase encodes, this might be a useful MR angiographic technique to be used in routine clinical practice. PMID- 12631514 TI - Presacral dermoid cyst with scanty fat component: usefulness of chemical shift and diffusion-weighted MR imaging. AB - We present a case of a dermoid cyst located at the presacral space. Although CT and conventional MR (T1- and T2-weighted images) demonstrated a non-specific cystic mass with little evidence of calcification or fat, chemical shift and diffusion-weighted MR imaging suggested the presence of small amount of fat and abundant keratinoid substance within the tumor, respectively. Pre-operative diagnosis of dermoid cyst was made. The tumor was surgically resected and the diagnosis was confirmed. Chemical shift and diffusion-weighted MR imaging are useful in the diagnosis of dermoid cyst with little evidence of calcification and fat. PMID- 12631513 TI - Perfusion changes of hepatic parenchyma due to infectious hepatobiliary disease: demonstration by perfusion CT. AB - We present two cases, in which perfusion CT demonstrated diffusely increased arterial and decreased portal perfusion of the liver parenchyma around liver abscess and infectious bilomas. These cases suggested that perfusion CT may be useful for a quantitative analysis of perfusion changes in infectious hepatobiliary diseases with good anatomical detail. PMID- 12631516 TI - Improving spatial resolution in SPECT with the combination of PSPMT based detector and iterative reconstruction algorithms. AB - This paper investigates the possibility of developing a SPECT system that combines the high spatial resolution of position sensitive photomultiplier tubes (PSPMTs) with the excellent performance of iterative reconstruction algorithms. A small field of view (FOV) camera based on a PSPMT and a pixelized scintillation crystal made of CsI(Tl) have been used for the acquisition of the projections. With the use of maximum likelihood expectation maximization (ML-EM) and ordered subsets expectation maximization (OSEM) slices of the object are obtained while three-dimensional (3D) reconstruction of the object is carried out using a modified marching cubes (MMC) algorithm. The spatial resolution of tomographic images obtained with the system was 2-3mm. The spatial resolution of a conventional system that uses filtered backprojection (FBP) for slices reconstruction was more than 9 mm. PMID- 12631515 TI - The glycine peak in brain diseases. AB - Glycine is an excitatory amino acid, a neurotransmitter for the brain. A recent experimental study by a 9.3T laboratory spectrometer identified the peak of pure glycine at 3.52 ppm, and in a clinical case this peak was demonstrated at 3.50 ppm by a 1.5 T clinical scanner. This study was undertaken to investigate the brain diseases having the glycine peak. An experiment with a 1.5 T clinical MRI unit was performed. Two grams of pure glycine was dissolved in 200 cc of distilled water and the solution was frozen, and proton MR spectroscopy (TR=1500 ms, TE=20 ms) was obtained. Nine patients with various diseases studied by two dimensional chemical shift spectroscopy (hybrid CSI) with TR=1500 ms, and TE=40 ms are included in the study. Ten normal cases were available for comparison. In the experiment with the clinical MRI unit, the glycine peak was centered at 3.50 ppm. The disease processes associated with distinct glycine peaks at 3.50 ppm included infarction, high-grade astrocytoma, megalencephalic leukoencephalopathy with cysts, Leigh's disease, adrenoleukodystrophy, congenital muscular dystrophy, Rasmussen's encephalitis, gliosis in neuronal migrational disorder, and hamartoma in tuberous sclerosis. None of the control cases displayed a glycine peak. In conclusion, glycine has a peak centered at 3.50 ppm in in vivo environments. It is distinct from the myoinositol peak. Detection of glycine in a wide variety of brain diseases ranging from infarction, tumor, leukoencephalopathies, infection to gliosis likely reflects presence of excitotoxic brain damage or a disturbance of neurotransmitting mechanisms in these conditions. PMID- 12631517 TI - Bilobed gallbladder diagnosed by oral cholecysto-CT. AB - Morphological anomalies of the gallbladder are rare and are characterized by a large variety of configurations depending on the size and degree of fusion of the lobes, and on the number and position of the cystic ducts. We presented a case of bilobed gallbladder incidentally detected during abdominal ultrasonography. Oral cholecystography and computed tomographic examination after oral cholecystography (Oral Cholecysto-CT) were performed to confirm this abnormality. This is a quite rare anomaly consisted of complete duplication of the fundus and partially duplication of the corpus and separated lobes both of which entered a single infundibulum and cystic duct. PMID- 12631518 TI - A double staining flow cytometric assay for the detection of steroid induced apoptotic leucocytes in common carp (Cyprinus carpio). AB - Steroid hormones play an important role in the regulation of the immune system through different ways. In this in vitro study, the effects of steroid hormones on the apoptosis of leucocytes were evaluated to understand the involvement of this process in the immunocompetence of common carp. Prior to the investigation, a double staining flow cytometric assay using fluorescein diacetate (FDA), which reacts with esterases of viable cells, and propidium iodide (PI), an acid dye that binds with nuclear DNA, was established. FDA and PI negative cells were regarded as apoptotic. The FDA-PI technique is comparable to the Annexin V-PI technique and can be used in the quantification of the apoptosis of fish leucocytes accurately. The results suggest that the disappearance of esterases and externalization of phosphatidylserine (PS) may be common to many apoptotic pathways. Cells collected from peripheral blood, spleen, head kidney, and thymus were cultured for 16 h either in the absence or presence of steroid hormones, i.e. cortisol (F), testosterone, 11-ketotestosterone, and estradiol-17beta, and analyzed by flow cytometry followed by the FDA-PI method. Results showed that F induced apoptosis in leucocytes from blood and other lymphoid organs suggesting the role of F as an immune regulator. The participation of sex steroids to the immunocompetence of carp was not found, since they did not induce apoptosis of leucocytes in any organ. PMID- 12631519 TI - Cloning and characterization of cDNAs for two distinct tumor necrosis factor receptor superfamily genes from Japanese flounder Paralichthys olivaceus. AB - Tumor necrosis factor receptor (TNFR) superfamily regulates diverse biologic functions, including cell proliferation, differentiation, and survival, in addition to providing costimulatory signals for programmed cell death or apoptosis. In this study, cDNA fragments for two distinct TNFR homologues were obtained from a Japanese flounder, Paralichthys olivaceus, cDNA library. Full length cDNAs of TNFR-1 and TNFR-2 homologues were obtained by using these cDNA fragments as probes. The cDNA for the Japanese flounder TNFR-1 homologue predicts a peptide of 395 amino acids that is 35% identical to the extracellular region of mouse TNFR-1, whereas the cDNA of the Japanese flounder TNFR-2 homologue predicts a peptide of 483 amino acids that is 40% identical to the extracellular region of human TNFR-2. The cytoplasmic domain contains a sequence that has the consensus motif of the death domain of the Japanese flounder TNFR-1 homologue. In a healthy fish, the mRNAs of both TNFR homologues were predominantly expressed in leukocytes, kidney, gill, and spleen. Expression of the Japanese flounder TNFR-1 homologue was induced in peripheral blood lymphocytes (PBLs) after stimulation with LPS (500 microg/ml) for 1 h, and TNFR-2 homologue was strongly induced in PBLs after stimulation with Con A (50 microg/ml) and PMA (0.35 microg/ml) for 3 h. The different expression patterns of the two distinct TNFR homologues may be critical in determining whether binding with TNF-alpha or TNF-beta have activating, proliferative, or apoptotic effects on target cells. PMID- 12631520 TI - Alternate forms of MHC class II-associated invariant chain are not produced by alternative splicing in rainbow trout (Oncorhynchus mykiss) but are encoded by separate genes. AB - A major limiting factor in understanding teleost major histocompatibility receptor function is the lack of knowledge about antigen presentation accessory molecules. We report here two cDNA clones encoding teleost versions of invariant chain and one encoding a related protein that may play a protease inhibition role in antigen presentation. The two invariant chain equivalents are similar to each other where they overlap, but differ in the presence or absence of a thyroglobulin domain. This domain is added to tetrapod invariant chain protein by alternative splicing but there was no evidence of alternative splicing of the two trout genes. Southern blotting confirmed that all three trout cDNAs are derived from single copy genes and Northern blotting indicated that they are expressed in antigen tissues. Thus the encoded proteins are probably involved in antigen presentation, but their expression is probably regulated in a manner different from tetrapods. PMID- 12631521 TI - LPS-stimulated expression of a tumor necrosis factor-alpha mRNA in primary trout monocytes and in vitro differentiated macrophages. AB - We have characterised the lipopolysaccharide (LPS) induction and stability of a tumor necrosis factor-alpha (TNFalpha) mRNA in both monocytes and mature macrophages cultured from trout head kidney. Macrophages differentiated in vitro significantly change in phenotypic appearance, phagocytic capacity and upon LPS exposure display a greatly increased capacity for trout TNFalpha (tTNF-alpha) mRNA expression over that of freshly isolated monocytes. The half-life of the tTNF-alpha mRNA in macrophages is approximately 40 min suggesting that control mechanisms for labile cytokine mRNAs in teleosts may be similar to that found in mammals. Our results suggest that in fish, terminally differentiated macrophages play a central and key role in TNF production as a response to infection. PMID- 12631522 TI - Macrophage migration inhibitory factor (MIF) of jawed and jawless fishes: implications for its evolutionary origin. AB - The macrophage migration inhibitory factor (MIF) is a cytokine produced by T lymphocytes and macrophages in response to inflammatory stimuli. We sequenced MIF cDNA clones of two jawless fishes, the sea lamprey (Petromyzon marinus) and the North Atlantic hagfish (Myxine glutinosa), as well as of the jawed (cichlid) fish Paralabidochromis chilotes. The fish MIF-encoding genes have the same exon-intron organization as the mammalian MIF genes and are present in one copy per haploid genome. Secondary and tertiary structure predictions suggest that the fish MIF proteins have a topology characteristic of the entire MIF-family of proteins. Phylogenetic analysis separates the known nematode members of the family into two groups, one having a sister group relationship with the mammalian D-dopachrome tautomerase (DDT) proteins and the other being related to vertebrate MIFs. It also reveals a high degree of convergent evolution among the members of the family. Finally, it suggests that the divergence of MIF and DDT occurred before the emergence of nematodes in metazoan evolution. PMID- 12631523 TI - Humoral immune response of carp (Cyprinus carpio) induced by oral immunization with liposome-entrapped antigen. AB - To study the value of liposomes as carriers of antigens for oral vaccination in fish, humoral immune responses were analyzed after immunizing carp (Cyprinus carpio) with liposome-entrapped bovine serum albumin (BSA) as a model antigen. Oral immunization of BSA (100 microg)-containing liposomes that were stable in carp bile induced significant antibody responses against BSA in serum as well as in intestinal mucus and bile. By contrast, no serum antibody responses were observed when fish were orally immunized with the same dose of BSA-containing unstable liposomes or BSA alone. BSA-specific antibody-secreting lymphocytes were detected in the spleen and head kidney of immunized fish. Furthermore, when we applied Vibrio cholerae toxin B subunit (CT-B)-conjugated liposomes containing BSA for oral immunization we found significant increases of anti-BSA antibodies in serum. Our results suggest that delivery systems using liposomes or liposomes with CT-B to the intestinal tract are essential for inducing effective humoral immune responses following oral vaccination. PMID- 12631524 TI - Oxidative burst mediated by toll like receptors (TLR) and CD14 on avian heterophils stimulated with bacterial toll agonists. AB - Toll-like receptors (TLRs) recognize pathogen-associated molecular patterns (PAMPs) such as lipopolysaccharide (LPS) and lipoteichoic acid (LTA), which are found in the cell walls of gram-negative and gram-positive bacteria, respectively. This study was conducted to determine if TLRs are present on chicken heterophils and if these receptors mediate oxidative burst. Heterophils isolated from neonatal chicks were exposed to gram-negative Salmonella enteritidis (SE), gram-positive Staphylococcus aureus (SA), SE-LPS, and SA-LTA and the oxidative burst quantitated by luminol-dependent chemiluminescence. SE, SA, SE-LPS, and SA-LTA stimulated a significant increase in oxidative burst from heterophils. Furthermore, we measured the inhibitory effects of polyclonal antibodies on rat CD14, human TLR2 and TLR4 on the oxidative burst of heterophils when stimulated with LPS and LTA. The data suggest that TLR2 and TLR4 mediate LPS stimulated oxidative burst while CD14 and TLR2 mediate LTA-stimulated oxidative burst in heterophils. This is the first report of PAMPs from gram-positive and gram-negative bacteria interacting with TLRs of avian heterophils. PMID- 12631525 TI - Immunological tolerance in chickens hatching from eggs injected with cell associated herpesvirus of Turkey (HVT). AB - Cell-associated herpesvirus of turkeys (HVT) was inoculated in ovo at various stages of incubation. Chickens hatching from these eggs were tested for anti-HVT antibodies by several serologic procedures including enzyme-linked immunosorbent assay, indirect immunofluorescence assays, and western blot. Viremic chickens that remained free of detectable antibodies were considered tolerant to HVT. Chickens exposed to HVT at embryonation day 14 or earlier had 6-33% incidence of tolerance. Tolerant chickens developed persistent HVT viremia. A preliminary challenge experiment provided circumstantial evidence that tolerant to HVT may be associated with reduced resistance to virulent Marek's disease virus. Tolerance to HVT did not influence the ability of the chickens to produce antibodies against an extraneous antigen or respond to a T cell mitogen. PMID- 12631526 TI - Activation marker expression on bovine peripheral blood gammadelta T cells during post-natal development and following vaccination with a commercial polyvalent viral vaccine. AB - Understanding the immunological function of bovine gammadelta T cells is essential for evaluating their role in the response to infectious agents and for determining the potential of targeting this population with vaccines. This study examined the age dependent changes of circulating CD2(+) and CD2(-) gammadelta T cells as well as differences in the expression of activation markers between these two populations. Changes in activation marker expression following vaccination with Vira Shield 5 are also discussed. CD62L was expressed on all CD2(-) gammadelta T cells but only a subset of CD2(+) gammadelta T cells and following vaccination there was a significant decrease in the percentage of CD2( )/CD62L(+) gammadelta T cells but not CD2(+)/CD62L(+) gammadelta T cells. Both CD2(-) and CD2(+) gammadelta T cells consistently expressed high levels of CD44. The majority of both CD2(-) and CD2(+) gammadelta T cells also expressed CD45R, however, more of the CD2(-) cells were CD45R(neg/lo). Following vaccination there was a significant decrease in the percentage of CD2(-) and CD2(+) gammadelta T cells that expressed CD44 and CD45R. These data indicate significant differences in activation expression on CD2(-) and CD2(+) gammadelta T cells, which adds to the growing evidence that there may be functional as well as phenotypic differences between these two populations of bovine gammadelta T cells. PMID- 12631527 TI - Multiple patterns of writing disorders in dementia of the Alzheimer type and their evolution. AB - This paper reports the results obtained from a writing task given to 23 Italian patients suffering from mild to moderate dementia of the Alzheimer's type (DAT). Spelling performance was tested with a task that taps the sub-word-level (spelling of regular words and nonwords), and the lexical route (spelling of regular and irregular words), in line with contemporary models of writing. Each patient's performance was classified according to the emergence of dissociated patterns of damage between regular words and nonwords and between regular and irregular words. The 23 DAT patients span the whole spectrum of dysgraphic taxonomy; five showed the characteristic pattern of impairment of surface dysgraphia, two showed the characteristics of phonological dysgraphia, while a mixed pattern (i.e. better performance on regular words compared to irregular words and regular nonwords) emerged in seven cases. Three patients presented undifferentiated writing disorders, two were completely agraphic, while four patients showed only minimal or no writing defects. The rate of dissociated impairments in the lexical and the sub-word-level routine is very similar to that observed after acute focal brain damage, which contradicts the hypothesis that degenerative brain damage selectively impairs writing performance along the lexical-semantic route. To test the hypothesis that surface sub-word-level processing abilities are affected only during the evolution of the disease, nine patients were tested longitudinally after an interval of 6-12 months. Once again, the data showed high variability across subjects, and do not seem to support involvement of the sub-word-level spelling routine only at a late stage in the development of the disease. PMID- 12631528 TI - Mixed emotions: alcoholics' impairments in the recognition of specific emotional facial expressions. AB - Facial expression recognition is a central feature of emotional and social behaviour and previous studies have found that alcoholics are impaired in this skill when presented with single emotions of differing intensities. The aim of this study was to explore biases in alcoholics' recognition of emotions when they were a mixture of two closely related emotions. The amygdala is intimately involved in encoding of emotions, especially those related to fear. In animals an increased number of withdrawals from alcohol leads to increased seizure sensitivity associated with facilitated transmission in the amygdala and related circuits. A further objective therefore was to explore the effect of previous alcohol detoxifications on the recognition of emotional facial expressions. Fourteen alcoholic inpatients were compared with 14 age and sex matched social drinking controls. They were asked to rate how much of each of six emotions (happiness, surprise, fear, sadness, disgust and anger) were present in morphed pictures portraying a mix of two of those emotions. The alcoholic group showed enhanced fear responses to all of the pictures compared to the controls and showed a different pattern of responding on anger and disgust. There were no differences between groups on decoding of sad, happy and surprised expressions. In addition the enhanced fear recognition found in the alcoholic group was related to the number of previous detoxifications. These results provide further evidence for impairment in facial expression recognition present in alcoholic patients. In addition, since the amygdala has been associated with the processing of facial expressions of emotion, particularly those of fear, the present data furthermore suggest that previous detoxifications may be related to changes within the amygdala. PMID- 12631529 TI - Effects of deep brain stimulation on prehensile movements in PD patients are less pronounced when external timing cues are provided. AB - It has been repeatedly demonstrated that the movements of patients with Parkinson's disease (PD) are less impaired when external timing cues are provided. This suggests that the basal ganglia, which are impaired in PD, are less involved in the control of externally timed movements. In the present study, we tested this hypothesis by contrasting the effect of deep brain stimulation (DBS) in the basal ganglia (more precisely, the internal globus pallidum) on internally versus externally timed movements. Our first movement task was a standard prehensile task involving a reach-to-grasp movement. In the externally timed condition, the target object was moving rapidly away from the subject; in the internally-timed condition, the target object was stationary. We found, that for most aspects of the prehensile movement the effect of DBS was less pronounced in the externally than in the internally timed condition. A similar reduction of the DBS effects in the externally-timed condition was also found for a second movement task, which required an isolated grasping movement. We conclude that the basal ganglia are significantly less involved in the control of externally timed movements. PMID- 12631530 TI - Hemispace differences in the visual perception of size in left hemiParkinson's disease. AB - The visual perception of size in different regions of external space was studied in Parkinson's disease (PD). A group of patients with worse left-sided symptoms (LPD) was compared with a group with worse right-sided symptoms (RPD) and with a group of age-matched controls on judgements of the relative height or width of two rectangles presented in different regions of external space. The relevant dimension of one rectangle (the 'standard') was held constant, while that of the other (the 'variable') was varied in a method of constant stimuli. The point of subjective equality (PSE) of rectangle width or height was obtained by probit analysis as the mean of the resulting psychometric function. When the standard was in left space, the PSE of the LPD group occurred when the variable was smaller, and when the standard was in right space, when the variable was larger. Similarly, when the standard rectangle was presented in upper space, and the variable in lower space, the PSE occurred when the variable was smaller, an effect which was similar in both left and right spaces. In all these experiments, the PSEs for both the controls and the RPD group did not differ significantly, and were close to a physical match, and the slopes of the psychometric functions were steeper in the controls than the patients, though not significantly so. The data suggest that objects appear smaller in the left and upper visual spaces in LPD, probably because of right hemisphere impairment. PMID- 12631531 TI - Rapid extraction of emotional expression: evidence from evoked potential fields during brief presentation of face stimuli. AB - Although the emotional expression of faces is believed to be accessed rapidly, previous ERP studies hardly found correlates of these processes. Here, we report findings from a study that investigated dichoptic binocular interaction using emotional face stimuli. Thirty-one subjects were briefly presented with schematic normal and scrambled faces (of neutral, positive, or negative expression) that occurred simultaneously in the left and right visual fields. Stimuli for both eyes could be congruent (control) or incongruent (dichoptic). Subjects decided which of the superimposed images in both hemi-fields appeared more "face-like" and during this task, the EEG was recorded from 30 channels. VEPs were analysed topographically according to the influence of the different experimental conditions (defined by presentation form, emotional expression, and location). Behavioural responses to the ambiguous dichoptic stimuli demonstrated a functional eye dominance not related to visual acuity and conventional eye preference. Electrophysiological data revealed three components with mean latencies of 85, 160, and 310 ms. Topography of the second component (equivalent to the face-related N170) differed in left-right and anterior-posterior direction compared with simple checkerboard stimuli. Dichoptic presentation caused reduced field strength of all three, and increased latency of the first component. Faces with negative expression yielded largest field strength of the second and third components. Besides that, emotional expression affected topography not only of late, but also the first component. This provides new evidence about the timing of perceptual processes related to facial expression, indicating that already VEP components occurring at 80-90 ms are sensitive to emotional content. PMID- 12631532 TI - Involvement of the medial temporal lobe in priming for new associations. AB - This study was addressed to the question of whether the medial temporal lobe (MTL) plays a critical role in implicit memory for new associations. Priming for new associations was examined in two different tasks in 18 patients with focal lesions all involving the MTL. In Experiment 1, following a study phase for pairs of unrelated words, subjects performed a perceptual identification task on old, recombined, and new pairs of words presented at brief exposure durations. In contrast to control subjects, and despite a normal level of item priming, the patients failed to show superior identification of the old pairs relative to the recombined pairs, the measure of associative priming. In Experiment 2, subjects engaged in speeded naming of the print color for previously studied words presented in the original color or in a different old color, and for unstudied words. Again, in contrast to control subjects and despite a normal level of item facilitation on color naming reaction time (RT), the patients failed to show priming for recently experienced new associations between words and colors. Explicit recognition memory by the patients was abnormal in both experiments. This study records an absence of priming for new associations, in two different tasks in which the nature of the stimuli was considerably different, in a large group of patients with lesions in the MTL. Although some previous research has reported significant associative priming in other tasks for patients with MTL lesions, the present results suggest that this region is critical for forming new associations of the types assessed here. PMID- 12631533 TI - Interhemispheric neural summation in the split brain: effects of stimulus colour and task. AB - Four split-brained subjects, two subjects with agenesis of the corpus callosum, and 14 normal subjects performed two tasks requiring responses to red or green disks, briefly presented either singly in the left visual field, singly in the right visual field, or simultaneously in both visual fields. In Experiment 1, simple reaction times to these stimuli, regardless of colour, were recorded (the Go-Both Task), and found to be faster to bilateral-redundant stimulus pairs, than to single stimuli. This so-called "redundancy gain" was much larger for acallosal or split-brained subjects than for normal subjects and exceeded the predictions of a race model, implying neural summation. Experiment 2 used the same stimuli, but subjects were required to respond only to stimuli of a designated colour (the Go/No-Go Task). Redundant target stimuli produced neural summation, while stimuli pairs that included a non-target stimulus did not. These results suggest that neural summation in the acallosal or split brain involves the convergence of response-associated activation, and that redundant sensory processes are not sufficient. PMID- 12631534 TI - Neural correlates of naming animals from their characteristic sounds. AB - The neural correlates of naming stimuli presented through the auditory modality have scarcely been studied. Using a PET experiment in 10 normal subjects, we began to address this issue by testing the hypothesis that naming animals from their characteristic sounds will engage bilateral primary auditory and auditory association cortices, bilateral early visual association cortices, left inferotemporal (IT) cortices, and left frontal operculum. Subjects listened to characteristic animal sounds (e.g. a rooster crowing), and named the animals making the sounds. When contrasted with a baseline task that involved saying up/down to the direction of pitch change in tone sequences, the naming task produced activation in mesial occipital cortices, the left ventral IT region, and the left frontal operculum. We interpret the activation in visual association cortices to reflect the process of retrieving conceptual knowledge (e.g. physical structure) pertinent to the animals being named, as in visual images. The left IT activation is interpreted to reflect activation of a mediation system for word retrieval, that operates to link conceptual knowledge retrieval to word production, and whose triggering is independent of the sensory modality in which a stimulus is presented. PMID- 12631535 TI - Different brain mechanisms mediate two strategies in arithmetic: evidence from Event-Related brain Potentials. AB - Participants were asked to verify if complex additions were smaller than 100 or not. Two hundred and forty arithmetic problems were presented, with half the problems being small-split problems (i.e. proposed sums were 2 or 5% away from 100) and half being large-split problems (i.e. proposed sums were 10 or 15% away from 100). Behavioral and ERPs data indicate that participants may use two different strategies to verify complex inequalities, a whole-calculation strategy for small-split problems and an approximate-calculation strategy for large-split problems. The choice between these two strategies occurred within 250 ms post stimulus presentation, and strategy execution was lateralized. Implications for our understanding of the brain mechanisms underlying arithmetic problem solving are discussed. PMID- 12631536 TI - Maternal occupational chemical exposures and biotransformation genotypes as risk factors for selected congenital anomalies. AB - In a case-control study using an assessment of occupational tasks by an industrial hygienist, the authors investigated whether women's occupational exposures increased risks of delivering infants with cleft palate (CP), cleft lip with or without cleft palate (CLP), conotruncal defects, or limb deficiencies. For CP and CLP, exposures were further considered in the presence/absence of infant genetic variants for glutathione-S-transferase M1, glutathione-S transferase T1, and N-acetyltransferases 1 and 2. The study included 1987-1989 California stillbirths and livebirths. Telephone interviews were conducted with mothers of 662 CLP and CP cases, 207 conotruncal defect cases, 165 limb deficiency cases, and 734 nonmalformed controls. Occupational tasks were assigned to a priori-defined exposure categories: 74 chemical groups and nine "end-use" chemical groups. Odds ratios of 1.5 or greater were observed for a small number of exposure-defect comparisons. Risks associated with end-use groups revealed odds ratios of 1.5 or greater for exposures to dyes and pigments (conotruncal and CP), propellants (CP), and insecticides (conotruncal and CP). Numerous odds ratios of 2.5 or greater were observed for combined effects of exposures and homozygous mutant genotypes, particularly for CP. Although potential associations were observed, most results suggested that maternal occupational chemical exposures did not contribute substantially to the occurrence of these anomalies in this California population. PMID- 12631537 TI - Prenatal exposure to low-level polychlorinated biphenyls in relation to mental and motor development at 8 months. AB - The relation between exposure to low levels of polychlorinated biphenyls (PCBs), a class of persistent organic pollutants, and cognitive and motor development in young children has been examined in several studies, and results have varied. The authors evaluated the association between prenatal exposure to PCBs and children's neurodevelopment using data from the Collaborative Perinatal Project. Pregnant women were enrolled from 1959 to 1965 from 12 sites across the United States. PCBs were measured in maternal serum taken during pregnancy. To measure children's mental and psychomotor development at 8 months of age, the authors administered the Bayley Scales of Infant Development (means, 87 (standard deviation, 15) and 88 (standard deviation, 18), respectively). Overall, they did not observe a relation between prenatal PCB exposure and children's mental or psychomotor scores (n = 1,207; multivariate adjusted beta = 0.1 point per micro g/liter increase of PCB, p = 0.71, and beta = 0.5, p = 0.14, respectively). The PCB-psychomotor score relation varied by study center (p < 0.05): The association was direct in some centers, inverse in others. This could not be attributed to variation in the timing or measurement of the child's neurodevelopment or analysis of PCBs because these were standardized across centers. The reasons for variation in results within this study and across other studies remain unclear. PMID- 12631538 TI - Scleroderma and solvent exposure among women. AB - Exposure to solvents has been reported to increase the risk of scleroderma. The authors investigated the relation between exposures to solvents in occupational and hobby settings and the development of scleroderma among women in a case control study with population-based controls in Michigan (1980-1991) and Ohio (1980-1992). A total of 660 cases and 2,227 frequency-matched controls were interviewed by telephone. Diagnoses of scleroderma were verified by medical records review. Paint thinners and removers were significantly associated with scleroderma both by self-report (odds ratio (OR) = 1.9, 95% confidence interval (CI): 1.4, 2.6) and after expert review (OR = 2.0, 95% CI: 1.5, 2.6). Other petroleum distillates (gasoline and mineral spirits) were not significantly associated with scleroderma after controlling for other correlated exposures in multivariable analyses. Trichloroethylene was associated with scleroderma both by self-report (OR = 2.0, 95% CI: 0.8, 4.8) and after expert review (OR = 1.9, 95% CI: 0.6, 6.6), but not significantly. Analyses by duration of exposure found that risk increased with the duration of use of any of the solvents (OR = 1.01/year of exposure, 95% CI: 1.01, 1.02), but there was no evidence of increasing risk with increasing duration of exposure for any specific solvent studied. In summary, exposures to paint thinners and removers were associated with scleroderma in women but showed no evidence of increasing risk with increasing duration. Exposures to other specific chlorinated and nonchlorinated hydrocarbon solvents were not clearly associated with scleroderma. PMID- 12631539 TI - Exposure to ambient fine particulate matter and primary cardiac arrest among persons with and without clinically recognized heart disease. AB - The authors studied the association between incidence of primary cardiac arrest and daily measures of fine particulate matter (/=20 years and were registered with an incident primary discharge diagnosis of non-Hodgkin's lymphoma between 1991 and 1998. This paper includes data on 211 cases and 800 controls individually matched on sex, age, community pharmacy, calendar time, and duration of follow-up. Conditional logistic regression analysis was used to evaluate the association between non Hodgkin's lymphoma and categories of cumulative drug use in days. In multivariate analyses, nonsignificant risk reductions were found for all drugs tested, and the negative association tended to increase with increasing duration of use. For women, the odds ratio for H(2) blockers was 0.29 (95% confidence interval: 0.12, 0.69) and for analgesics was 0.40 (95% confidence interval: 0.22, 0.71). Results support an inverse association between occurrence of non-Hodgkin's lymphoma and use of H(2) blockers and analgesics among women, and they warrant confirmation in larger studies. PMID- 12631541 TI - Body mass index in adolescence in relation to total mortality: 32-year follow-up of 227,000 Norwegian boys and girls. AB - A large number of studies have explored the relation between body mass index (BMI) and mortality in adults. The relation between BMI in adolescence and mortality has been investigated to a lesser extent. It has been suggested that all-cause mortality is elevated among those who were overweight during adolescence, but the limitation of previous studies has been study size. The present study explored this relation in a Norwegian cohort of 227,003 boys and girls, aged 14-19 years, whose height and weight were measured during tuberculosis screening in 1963-1975. These persons were followed for an average of 31.5 years (about 7.2 million person-years). A total of 7,516 deaths were registered. Multivariate Cox proportional hazards regression models were used in the analyses. An increasing risk of death by increasing BMI in adolescence was observed. Mortality among males whose baseline BMI was between the 85th and 95th percentiles and above the 95th percentile in the US reference population was 30% and 80% higher, respectively, than that among those whose baseline BMI was between the 25th and 75th percentiles. The corresponding rates among females were 30% and 100%. The excess mortality among adolescents whose BMI was high was not clearly manifested before they reached their thirties. Hence, BMI in adolescence is predictive of adult mortality. PMID- 12631543 TI - Effects of climate on variability in Lyme disease incidence in the northeastern United States. AB - Numbers of reported Lyme disease cases have increased dramatically over the past decade in the northeastern United States, but the year-to-year variability is sizable (average standard deviation approximately 30% of the mean). An improved understanding of the causes of such variability would aid in prevention and control of the disease, which is transmitted by a spirochete carried in the "black-legged" tick, Ixodes scapularis. In this study, the variability in reported Lyme disease incidence between 1993 and 2001 in seven northeastern US states was analyzed as an outcome of weather variability. For all seven states analyzed, significant (p < 0.05) positive relations were found for the correlation of early summer disease incidence with the June moisture index (Palmer Hydrological Drought Index) in the region 2 years previously. The correlations may reflect enhanced nymph tick survival in wetter conditions. Few significant relations were found with same-year moisture index, which suggests that moisture has a greater effect on nymph tick survival following the insect's blood meal than before. In some states, significant correlations were observed related to warmer winter weather a year and a half prior to disease incidence, which may have been due to higher survival and activity levels of the white footed mouse, the main host for Lyme disease-infected ticks. PMID- 12631542 TI - Anatomic distribution of sensory symptoms in the hand and their relation to neck pain, psychosocial variables, and occupational activities. AB - To explore whether different distributions of numbness and tingling in the hand can be usefully distinguished in epidemiologic studies of disorders such as carpal tunnel syndrome, the authors used a postal questionnaire, an interview, and a physical examination to collect information about risk factors, symptoms, and signs from a general population sample of 2,142 adults in Southampton, England, during 1998-2000. The authors distinguished six distributions of numbness and tingling and compared their associations with other clinical findings and with known risk factors for upper limb disorders. Distinctive relations were found for symptoms that involved most of the palmar surface of the first three digits but not the dorsum of the hand or the little finger. Such symptoms were more often associated with positive Phalen's and Tinel's tests and, unlike other categories of sensory disturbance, were not related to neck pain or restriction of neck movement. They also differed in showing no association with lower vitality or poorer mental health but an association with repeated wrist and finger movements at work. These findings suggest that, in the classification of numbness and tingling of the hand, it may be useful to distinguish symptoms that involve most of the sensory distribution of the median nerve but not other parts of the hand. PMID- 12631544 TI - Validation of self-reported cancers in the California Teachers Study. AB - Self-reported cancer data from the California Teachers Study were validated by using California Cancer Registry data. The California Teachers Study cohort consists of 133,479 active and retired California teachers. In 1995-1996, data from a mailed questionnaire were linked to the California Cancer Registry data. Sensitivity and specificity of 11 types of cancer were calculated. Multivariate analyses were conducted to evaluate correlates of false-positive and false negative reporting. Sensitivities showed great variation by cancer site. The highest sensitivities were observed for breast (96.4%) and thyroid (92.9%) cancers, whereas the lowest sensitivities were those for cervical (44.3%), endometrial (69.1%), and other skin (53.6%) cancers. The sensitivities for in situ cancers (at the time of diagnosis) were considerably lower than those for invasive cancers in about half of the cancer types surveyed. The specificities for individual cancer sites ranged from 90% to 99%; the highest were those for lung cancer, leukemia, and Hodgkin's disease (all 99.9%). The lowest specificity was for other skin cancer (90.2%). In situ stage at diagnosis and older age were significantly associated with false-positive reporting. Age and non-White race were associated with false-negative reporting. These findings suggest that the feasibility of using self-reported data without verification in epidemiologic studies of cancer varies by site. PMID- 12631545 TI - Training, quality assurance, and assessment of medical record abstraction in a multisite study. AB - Clinical studies using medical record review should include careful training and quality assurance methods to enhance the reliability and validity of data obtained from the records. Because of time and budget constraints, comprehensive assessments of data quality and reliability, including masking of medical record abstractors, are not always possible. This paper describes the abstractor training and quality control methods and results of a masked medical record review study. The medical record review study was carried out within a larger multisite study of the effectiveness of screening mammography in preventing breast cancer mortality with an observation period within 1983 and 1993, with mortality follow-up through 1998. An eight-step program was developed to train medical record abstractors and monitor the quality of their work. A key follow-up component to the training protocol was a 5% reabstraction of medical records (n = 160), masked and reviewed by a second abstractor. High agreement was found between initial (unmasked) abstractors and masked abstractors for all key exposure variables (kappa ranged from 0.76 to 0.91), with no evidence of biased directionality by unmasked reviewers. Rigorous ongoing training programs for medical record abstractors provide assurance of good quality control in large multisite studies. Additionally, a masking study with a subsample of subjects may be a feasible and cost-effective alternative to the time- and cost-intensive methodological approach of masking all medical records. PMID- 12631546 TI - Can we rely on retrospective pain assessments? AB - The objective of this paper was to study whether subjects in a workplace setting are able to assess the intensity of musculoskeletal pain retrospectively for a period of 3 months. The intensity of average pain and maximum pain in eight anatomic regions was assessed on a numeric rating scale. The results of 12 consecutive weekly pain recordings were compared with a final retrospective assessment of pain intensity covering the same 3-month period (119 subjects). The degree of agreement was good or excellent. The subjects were able to distinguish between the worst complaints and average complaints, and the subjective perception of aggravations or improvements corresponded to the direction of a change in score. Current complaint status slightly influenced the memory of pain. The reproducibility of the questionnaire was also assessed and showed kappa coefficients between 0.44 and 0.91 (36 subjects). The results suggest that subjects are able to accurately recall and rate the severity of pain or discomfort for a period of 3 months. These findings are of practical importance in epidemiologic studies, because they imply that retrospective reports on pain intensity are sufficiently reliable. PMID- 12631547 TI - Contribution of follow-up of nonresponders to prevalence and risk estimates: a Norwegian respiratory health survey. AB - Achieving a high response rate is often expensive and time consuming. Does an extensive survey effort change prevalence estimates and exposure-disease relations? In 1998-1999, the authors conducted a population-based respiratory health survey in two Norwegian counties (Oslo, n = 20,000; Hordaland, n = 5,000) of a random sample of the adult population aged 15-70 years. A postal questionnaire was mailed, with as many as two reminder letters. A 25% random sample of postal nonresponders was contacted by ordinary or cellular telephone. Cumulative response rates after the first mailing, first reminder, second reminder, and telephone follow-up were 42.7%, 60.7%, 68.3%, and 79.9%, respectively. Compared with initial postal responders, responders at later stages were younger, and more were male. Late responders had a lower educational level, were more often smokers, and were occupationally exposed to dusts or gases more frequently. After the authors adjusted for these factors, late responders were found to have less asthma, attacks of breathlessness, and hay fever. Hardly any changes in prevalences and odds ratios were noted when initial responders were compared with all responders. The additional contribution of sending reminder letters and conducting a telephone follow-up to prevalence estimates and the exposure-disease relation was small. A single letter would have produced nearly the same prevalence estimates and risk factor associations. PMID- 12631548 TI - Re: "Invited commentary: unexplained health problems after Gulf War service- finding answers to complex questions". PMID- 12631550 TI - Re: "Dose specific meta-analysis and sensitivity analysis of the relation between alcohol consumption and lung cancer risk". PMID- 12631551 TI - "Step" vs. "dynamic" autoregulation: implications for susceptibility to hypertensive injury. AB - Renal autoregulatory (AR) mechanisms provide the primary protection against transmission of systemic pressures, and their impairment is believed to be responsible for the enhanced susceptibility to hypertensive renal damage in renal mass reduction (RMR) models. Assessment of AR capacity by the "step" change methodology under anesthesia was compared with that by "dynamic" methods in separate conscious control Sprague-Dawley rats and after uninephrectomy (UNX) and (3/4) RMR (RK-NX) (n = 7-10/group). Substantially less AR capacity was seen by the dynamic vs. the step methodology in control rats. Moreover, dynamic AR capacity did not differ among controls, UNX, and RK-NX rats (fractional gain in admittance approximately 0.4-0.5 in all groups at frequencies in the range of 0.0025-0.025 Hz). By contrast, significant impairment of step AR was seen in RK NX vs. control or UNX rats (AR indexes 0.7 +/- 0.1 vs. 0.1 +/- 0.02 and 0.2 +/- 0.04, respectively, P < 0.01). We propose that the step and dynamic methods evaluate the renal AR responses to different components of blood pressure (BP) power with the step AR assessing the ability to buffer large changes in average BP (DC power), whereas the present "dynamic" methods assess the AR ability to buffer slow BP fluctuations (<0.25 Hz) superimposed on the average BP (AC power), a substantially smaller component of total BP power. We further suggest that step but not dynamic AR methods as presently performed provide a valid index of the underlying susceptibility to hypertensive glomerular damage after RMR. PMID- 12631552 TI - NO dependency of RBF and autoregulation in the spontaneously hypertensive rat. AB - In the spontaneously hypertensive rat (SHR), renal blood flow (RBF) has been reported to be very dependent on nitric oxide (NO); however, autoregulation is normal, albeit shifted to higher perfusion pressures. To test the hypothesis that in the SHR NO dependency of RBF autoregulation is diminished, we investigated RBF autoregulation in anesthetized young male SHR and normotensive Wistar-Kyoto (WKY) rats before and during acute intravenous NO synthase (NOS) inhibition with N(omega)-nitro-L-arginine (L-NNA) and urinary excretion of nitrate plus nitrite (U(NOx)V) at different renal perfusion pressures (RPP). Under baseline conditions, SHR had higher mean arterial pressure (147 +/- 4 mmHg) and renal vascular resistance (16 +/- 1 U) than WKY (105 +/- 4 mmHg and 10 +/- 0.5 U, respectively, P < 0.05). RBF was similar (9.4 +/- 0.5 vs. 10.3 +/- 0.1 ml x min( 1)x g kidney wt(-1)). Acute NOS blockade increased mean arterial pressure similarly, but there was significantly more reduction in RBF and hence an enhanced increase in renal vascular resistance in SHR (to 36 +/- 3 vs. 17 +/- 1 U in WKY, P < 0.001). The renal vasculature of SHR is thus strongly dependent on NO in maintaining basal RBF. The lower limit of autoregulation was higher in SHR than WKY in the baseline situation (85 +/- 3 vs. 71 +/- 2 mmHg, P < 0.05). Acute L-NNA administration did not decrease the lower limit in the SHR (to 81 +/- 3 mmHg, not significant) and decreased the lower limit to 63 +/- 2 mmHg (P < 0.05) in the WKY. The degree of compensation as a measure of autoregulatory efficiency attained at spontaneous perfusion pressures was comparable in SHR vs. WKY but with a shift of the curve toward higher perfusion pressures in SHR. Acute NOS blockade only increased the degree of compensation in WKY. Remarkably, U(NOx)V was significantly lower at spontaneous RPP in SHR. After reduction of RPP, the observed decrease in U(NOx)V was significantly more pronounced in WKY than in SHR. In conclusion, the renal circulation in SHR is dependent on high levels of NO; however, the capacity to modulate NO in response to RPP-induced changes in shear stress seems to be limited. PMID- 12631554 TI - Expression of the prostaglandin F receptor (FP) gene along the mouse genitourinary tract. AB - PGF(2alpha) is one of the major prostanoids produced by the kidney. The cellular effects of PGF(2alpha) are mediated by a G protein-coupled transmembrane receptor designated the FP receptor. Both in situ hybridization and beta-galactosidase knocked into the endogenous FP locus were used to determine the cellular distribution of the mouse FP receptor. Specific labeling was detected in the kidney, ovary, and uterus. Abundant FP expression in ovarian follicles and uterus is consistent with previous reports of failed parturition in FP-/- mice. In the kidney, coexpression of the mFP mRNA with the thiazide-sensitive cotransporter defined its expression in the distal convoluted tubule (DCT). FP receptor was also present in aquaporin-2-positive cortical collecting ducts (CCD). No FP mRNA was detected in glomeruli, proximal tubules, or thick ascending limbs. Intrarenal expression of the FP receptor in the DCT and CCD suggests an important role for the FP receptor regulating water and solute transport in these segments of the nephron. PMID- 12631555 TI - The future of GI and liver research: editorial perspectives: I. Visions of epithelial research. AB - Epithelial cells are gatekeepers that sit at the interface between two compartments. By controlling the flow of molecules and information between two compartments, epithelial cells provide unique benefit to the body. This article provides a brief appraisal of our current knowledge about the functions of gastrointestinal epithelial cells as a functionally diverse set of cells mediating transepithelial transport and as a continually renewing layer of cells. The convergence of new methodologies in laser capture microdissection, microarray analyses, microscopic analyses, and generation of mutant animals provides an exciting template for future research. PMID- 12631553 TI - Urinary excretion of viable podocytes in health and renal disease. AB - The loss of glomerular visceral epithelial cells (podocytes) has been associated with the development of glomerular sclerosis and loss of renal function. Viability of podocytes recovered from urine of subjects with glomerular disease and of healthy controls was investigated by propidium iodide exclusion and TUNEL staining. Podocyte loss was quantified by cytospin. The growth behavior in culture of urinary cells and their expression of specific markers were examined. The majority of urinary podocytes are viable, although apoptosis occurs in about one-half of the cells. Patients with active glomerular disease excreted up to 388 podocytes/mg creatinine, whereas healthy controls and patients with quiescent disease generally excreted <0.5 podocytes/mg creatinine. The identity of cultured cells was confirmed by their morphology, growth behavior, and expression of podocyte-specific markers. The difference in growth behavior between healthy controls and subjects with active glomerular disease suggests that in active disease viable podocytes detach from the glomerular tuft due to local environmental factors rather than defects in the podocytes per se, whereas in healthy individuals mostly senescent podocytes are shed. PMID- 12631556 TI - Bile acid regulation of hepatic physiology: III. Regulation of bile acid synthesis: past progress and future challenges. AB - Bile acids, amphipathic detergent-like molecules synthesized from cholesterol, are highly conserved by means of enterohepatic circulation. They participate in the generation of bile flow and biliary lipid secretion and also promote absorption of fat-soluble vitamins and lipids. Conversion of cholesterol to bile acids represents a quantitatively important route to eliminate cholesterol from the body. Regulation of bile acid synthesis involves a complex and interrelated group of transcription regulators that link bile acid synthesis to cholesterol and fatty acid metabolism. Targeting key steps of bile acid synthetic pathways as well as the metabolic network that maintains homeostatic levels of lipids should provide exciting novel opportunities for the treatment of cardiovascular and liver diseases. PMID- 12631558 TI - Deglutitive inhibition affects both esophageal peristaltic amplitude and shortening. AB - Deglutitive inhibition attenuates ongoing esophageal contractions if swallows are separated by short time intervals. This study aimed to determine whether esophageal shortening, mediated by longitudinal muscle, was similarly affected. Eight healthy subjects with two distal esophageal segments demarcated by mucosal clips and manometric recording sites positioned within those segments underwent concurrent manometry and fluoroscopy. Peristaltic amplitude and change in distal segment lengths were quantified during single swallows, paired swallows separated by progressively prolonged intervals, and a series of rapid repetitive swallows. During grouped swallows, deglutitive inhibition with complete attenuation of both the manometric contraction and segment shortening was evident with short-interval swallows and rapid-sequence swallows. No inhibition of either was evident with long-interval pairs. With intermediate interswallow intervals, the occurrence and degree of deglutitive inhibition between peristaltic amplitude and segment shortening were closely correlated. Deglutitive inhibition affects both the longitudinal and circular muscle layers of the esophageal wall, and the occurrence of inhibition evident in one layer is strongly correlated with the other. PMID- 12631557 TI - Effects of a kappa-opioid agonist, asimadoline, on satiation and GI motor and sensory functions in humans. AB - To compare the effects of the kappa-opioid agonist asimadoline and placebo on visceral sensation and gastrointestinal (GI) motor functions in humans, 91 healthy participants were randomized in a double-blind fashion to 0.15, 0.5, or 1.5 mg of asimadoline or placebo orally twice a day for 9 days. We assessed satiation (nutrient drink test), colonic compliance, tone, perception of colonic distension (barostat), and whole gut transit (scintigraphy). Treatment effect was assessed by analysis of covariance. Asimadoline increased nutrient drink intake (P = 0.03). Asimadoline decreased colonic tone during fasting (P = 0.03) without affecting postprandial colonic contraction, compliance, or transit. Gas scores in response to colonic distension were decreased with 0.5 mg of asimadoline at low levels (8 mmHg above operating pressure) of distension (P = 0.04) but not at higher levels of distension. Asimadoline at 1.5 mg increased gas scores at 16 mmHg of distension (P = 0.03) and pain scores at distensions of 8 and 16 mmHg (P = 0.003 and 0.03, respectively) but not at higher levels of distension. Further studies of this compound in diseases with altered satiation or visceral sensation are warranted. PMID- 12631559 TI - Therapeutic effect of anti-OX40L and anti-TNF-alpha MAbs in a murine model of chronic colitis. AB - Interaction of OX40 (CD134) on T cells with its ligand (OX40L) on antigen presenting cells has been implicated in pathogenic T cell activation. This study was performed to explore the involvement of OX40/OX40L in the development of T cell-mediated chronic colitis. We evaluated both the preventive and therapeutic effects of neutralizing anti-OX40L MAb on the development of chronic colitis in SCID mice induced by adoptive transfer of CD4(+)CD45RB(high) T cells as an animal model of Crohn's disease. We also assessed the combination of anti-OX40L and anti TNF-alpha MAbs to improve the therapeutic effect. Administration of anti-OX40L MAb markedly ameliorated the clinical and histopathological disease in preventive and therapeutic protocols. In vivo treatment with anti-OX40L MAb decreased CD4(+) T cell infiltration in the colon and suppressed IFN-gamma, IL-2, and TNF-alpha production by lamina propria CD4(+) T cells. The combination with anti-TNF-alpha MAb further improved the therapeutic effect by abolishing IFN-gamma, IL-2, and TNF-alpha production by lamina propria CD4(+) T cells. Our present results suggested a pivotal role of OX40/OX40L in the pathogenesis of T cell-mediated chronic colitis. The OX40L blockade, especially in combination with the TNF-alpha blockade, may be a promising strategy for therapeutic intervention of Crohn's disease. PMID- 12631560 TI - TRPC5 as a candidate for the nonselective cation channel activated by muscarinic stimulation in murine stomach. AB - We investigated which transient receptor potential (TRP) channel is responsible for the nonselective cation channel (NSCC) activated by carbachol (CCh) in murine stomach with RT-PCR and the electrophysiological method. All seven types of TRP mRNA were detected in murine stomach with RT-PCR. When each TRP channel was expressed, the current-voltage relationship of mTRP5 was most similar to that recorded in murine gastric myocytes. mTRP5 showed a conductance order of Cs(+) > K(+) > Na(+), similar to that in the murine stomach. With 0.2 mM GTPgammaS in the pipette solution, the current was activated transiently in both NSCC in the murine stomach and the expressed mTRP5. Both NSCC activated by CCh in murine stomach and mTRP5 were inhibited by intracellularly applied anti-G(q/11) antibody, PLC inhibitor U-73122, IICR inhibitor 2-aminoethoxydiphenylborate, and nonspecific cation channel blockers La(3+) and flufenamate. There were two other unique properties. Both the native NSCC and mTRP5 were activated by 1-oleoyl-2 acetyl-sn-glycerol. Without the activation of NSCC by CCh, the NSCC in murine stomach was constitutively active like mTRP5. From the above results, we suggest that mTRP5 might be a candidate for the NSCC activated by ACh or CCh in murine stomach. PMID- 12631561 TI - Gastric atrial natriuretic peptide regulates endocrine secretion in antrum and fundus of human and rat stomach. AB - Atrial natriuretic peptide (ANP) is present in gastric mucosa and preferentially binds to two subtypes of natriuretic peptide receptors (NPR), NPR-A and NPR-C. The present study examines the role of endogenous ANP in regulating endocrine secretion in rat and human stomachs. NPR-A protein expression and transcripts were identified in rat antral and fundic mucosa by Western blot and RT-PCR. In superfused rat and human antral and fundic segments, ANP (0.1 pM to 0.1 microM) caused a concentration-dependent increase in somatostatin secretion. In antrum, this was accompanied by a decrease in gastrin, and in fundus, this was accompanied by a decrease in histamine secretion. Changes in gastrin and histamine secretion reflected changes in somatostatin secretion and were abolished by somatostatin antibody. The NPR-A receptor antagonist anantin 1) inhibited basal somatostatin secretion and 2) abolished the somatostatin, gastrin, and histamine responses to ANP. We conclude that endogenous ANP, acting via the NPR-A receptor, stimulates somatostatin secretion from both antrum and fundus of rat and human stomach. Stimulation of somatostatin secretion is coupled to inhibition of gastrin secretion in the antrum and inhibition of histamine secretion in the fundus. PMID- 12631562 TI - Dissociating the roles of the rostral anterior cingulate and the lateral prefrontal cortices in performing two tasks simultaneously or successively. AB - A fundamental question about the nature of cognitive control is whether performing two tasks successively or simultaneously activates distinct brain regions. To investigate this question, we designed a functional magnetic resonance imaging (fMRI) study that compared task-switching and dual-task performance. The results showed that performing two tasks successively or simultaneously activated a common prefronto-parietal neural network relative to performing each task separately. More importantly, we found that the anterior cingulate and the lateral prefrontal cortices were differently activated in dual task and task-switching situations. When performing two tasks simultaneously, as compared to performing them in succession, activation was found in the rostral anterior cingulate cortex. In contrast, switching between two tasks, relative to performing them simultaneously, activated the left lateral prefrontal cortex and the bilateral intra-parietal sulcus region. We interpret these results as indicating that the rostral anterior cingulate cortex serves to resolve conflicts between stimulus-response associations when performing two tasks simultaneously, while the lateral prefrontal cortex dynamically selects the neural pathways needed to perform a given task during task switching. PMID- 12631564 TI - Different inhibitory synaptic input patterns in excitatory and inhibitory layer 4 neurons of ferret visual cortex. AB - The synaptic mechanisms underlying the generation of orientation and direction selectivity in layer 4 of the primary visual cortex are still largely unclear. Previous in vivo work has shown that intra-cortical inhibition plays a major role in generating the properties of orientation and direction selectivity. Excitatory and inhibitory cortical neurons differ in their receptive field properties: excitatory neurons tend to be orientation- and direction-selective, inhibitory neurons tend to be orientation-, but not direction-selective. Here we have compared the relationship between direction preference maps recorded in vivo and synaptic input maps recorded in vitro from excitatory and inhibitory stellate cells in layer 4 of ferret visual cortex. Our goal was to test whether the differences in direction tuning between these cell populations might result from different inhibitory connectivity patterns. We found that excitatory neurons, which are direction tuned in vivo, receive approximately 50% of their inhibitory inputs from cortical regions of opposite direction preference whereas inhibitory cells, which are not or poorly direction tuned, receive only very few inputs from regions of opposite direction preference. This confirms that inhibitory connections arising in cortical regions of opposite direction preference may be required to create or strengthen direction tuning in their target neurons. Thus, differences in intracortical inhibitory circuit patterns may underlie the differences in receptive field properties observed between excitatory and inhibitory neurons in vivo. PMID- 12631563 TI - Functional MRI studies of human visual motion perception: texture, luminance, attention and after-effects. AB - Motion of an object is thought to be perceived independently of the object's surface properties. However, theoretical, neuropsychological and psychophysical observations have suggested that motion of textures, called 'second-order motion', may be processed by a separate system from luminance-based, or 'first order', motion. Functional magnetic resonance imaging (fMRI) responses during passive viewing, attentional modulation and post-adaptation motion after-effects (MAE) of these stimuli were measured in seven retinotopic visual areas (labeled V1, V2, V3, VP, V4v, V3A and LO) and the motion-sensitive area MT/MST (V5). In all visual areas, responses were strikingly similar to motion of first- and second-order stimuli. These results differ from a prior investigation, because here the motion-specific responses were isolated. Directing attention towards and away from the motion elicited equivalent response modulation for the two types. Dramatic post-adaptation (MAE) differences in perception of the two stimuli were observed and fMRI activation mimicked perceptual changes, but did not reveal the processing differences. In fact, no visual area was found to respond selectively to the motion of second-order stimuli, suggesting that motion perception arises from a unified motion detection system. PMID- 12631565 TI - Hemispheric surface area: sex, laterality and age effects. AB - The surface area of the cortex is theoretically important but more complicated to measure than cortical volume. Its theoretical importance is primarily due to a close relationship between the surface area (not the volume) of the cortex at any region and the number of neurons in that region. The surface of the cortex in humans is highly convoluted and this creates difficulties in measuring its surface area. We report here on a straightforward extension of stereologic methods to magnetic resonance images that is simple, efficient and elegant. We studied the method's reliability and the relationship of measurement error to biological variation and suggest a simple method, using variance components analysis, for quantifying all the relevant parameters for efficient design of experiments. We applied this method to a small sample of 15 healthy young male subjects, matched individually on age and parental socioeconomic status to 15 healthy female subjects and measured the surface areas of both cerebral hemispheres. We found no evidence for gender differences or asymmetry in this small study, unlike a previous report using post-mortem material. PMID- 12631567 TI - Modulation of neural activity during object naming: effects of time and practice. AB - Repeated exposure to objects improves our ability to identify and name them, even after a long delay. Previous brain imaging studies have demonstrated that this experience-related facilitation of object naming is associated with neural changes in distinct brain regions. We used event-related functional magnetic resonance imaging (fMRI) to examine the modulation of neural activity in the object naming system as a function of experience and time. Pictures of common objects were presented repeatedly for naming at different time intervals (1 h, 6 h and 3 days) before scanning, or at 30 s intervals during scanning. The results revealed that as objects became more familiar with experience, activity in occipitotemporal and left inferior frontal regions decreased while activity in the left insula and basal ganglia increased. In posterior regions, reductions in activity as a result of multiple repetitions did not interact with time, whereas in left inferior frontal cortex larger decreases were observed when repetitions were spaced out over time. This differential modulation of activity in distinct brain regions provides support for the idea that long-lasting object priming is mediated by two neural mechanisms. The first mechanism may involve changes in object-specific representations in occipitotemporal cortices, the second may be a form of procedural learning involving a reorganization in brain circuitry that leads to more efficient name retrieval. PMID- 12631566 TI - Significance of dopamine transmission in the rat medial prefrontal cortex for conditioned fear. AB - Previous studies have demonstrated activation of dopamine transmission in the medial prefrontal cortex (mPFC) by conditioned fear stimuli. Therefore, the present study investigated the functional significance of mPFC dopamine for a conditioned fear response to a tone. We examined the effects of inhibition or stimulation of mPFC dopamine transmission by local microinfusion of the D1/D2 receptor antagonist cis-flupenthixol or the indirect dopamine receptor agonist D amphetamine, respectively, in a classical fear-conditioning paradigm in Wistar rats. Rats received tone-shock pairings and were later exposed to the tone alone. Freezing was used as measure of conditioned fear. Presence of the drugs in the mPFC during the tone-shock pairings did not affect freezing during subsequent presentation of the tone alone. However, when cis-flupenthixol and D-amphetamine were present in the mPFC during presentation of the tone alone, freezing to the tone was reduced. We demonstrated that the decreased freezing could be explained neither by state dependency nor infusion-induced alterations in activity. Our data indicate that mPFC dopamine transmission is important for the retrieval/expression, but not the formation, of conditioned fear. The reduction of conditioned fear by prefrontal infusion of both cis-flupenthixol and D amphetamine may reflect normal expression of conditioned fear requires an optimal level of mPFC dopamine activity. PMID- 12631568 TI - Contralateral coding of imagined body parts in the superior parietal lobe. AB - In monkeys, neurons in the superior parietal lobe (area 5) code for spatial position of contralateral body parts by combining visual and somatosensory signals. Using a modified version of the classical mental rotation task, we were able to demonstrate that in humans activation in the contralateral superior parietal lobe could be evoked when mental rotation was combined with motor imagery of hands. These findings show that even in the absence of visual and somatosensory input, information provided by motor imagery suffices to induce contralateral superior parietal lobe monitoring of the imagined limb configuration. This constitutes an important prerequisite for effective imagined motor practice that can be used to improve actual motor performance. PMID- 12631569 TI - Coordination of actions and habits in the medial prefrontal cortex of rats. AB - As animals learn novel behavioural responses, performance is maintained by two dissociable influences. Initial responding is goal-directed and under voluntary control, but overtraining of the same response routine leads to behavioural autonomy and the development of habits that are no longer voluntary or goal directed. Rats normally show goal-directed performance after limited training, indexed by sensitivity to changes in the value of reward, but this sensitivity to goal value is lost with extended training. Rats with selective lesions of the prelimbic medial prefrontal cortex showed no sensitivity to goal value after either limited or extended training, whereas rats with lesions of the infralimbic region of the medial prefrontal cortex showed the opposite pattern of deficit, a marked sensitivity to goal value after both limited and extended training. This double-dissociation suggests that the prelimbic region is responsible for voluntary response performance and the infralimbic cortex mediates the incremental ability of extended training to override this goal-directed behaviour. PMID- 12631570 TI - Carbachol-induced network oscillations in the intact cerebral cortex of the newborn rat. AB - In mature cortex, activation of the cholinergic system induces oscillatory network activity and facilitates synaptic plasticity. We used an in vitro preparation of the intact cerebral cortex and cortical slices of the neonatal rat to study carbachol (CCh, >or=30 micro M)-induced network oscillations during the early postnatal period. Multi-site extracellular recordings revealed CCh-induced transient beta oscillations with an average duration of 4.6 +/- 0.2 s, amplitude of 123 +/- 7.4 microV and frequency of 17.7 +/- 0.5 Hz. These oscillations propagated uniformly at 0.5-1.5 mm/s over the cortex and were reversibly blocked by tetrodotoxin (TTX) and atropine, indicating that they depended on action potentials and activation of muscarinic receptors. The activity was not blocked by bicuculline methiodide or gabazine, but was reversibly abolished by kynurenic acid, indicating that activation of glutamate receptors, but not GABA-A receptors, was required. CNQX caused a significant decrease in the power of the Fourier frequency spectrum of the CCh-induced oscillations and CPP or MK-801 completely blocked the activity, indicating a contribution of AMPA/kainate receptors and an essential role of NMDA receptors. Oscillations were synchronized between sites separated horizontally by approximately 1 mm and for delays of 2-8 ms. Synchronized activity between neighboring recording sites was very stable over repeated applications of CCh. Whole-cell recordings from morphologically identified pyramidal neurons in the intact cortex revealed a close temporal correlation between CCh-induced membrane oscillations and local field potential recordings. In contrast, CCh-induced oscillations recorded in coronal neocortical slices were smaller in amplitude and frequency, suggesting that a widespread network of intracortical axonal connections is required for their generation. PMID- 12631571 TI - Very slow activity fluctuations in monkey visual cortex: implications for functional brain imaging. AB - We examined fluctuations in band-limited power (BLP) of local field potential (LFP) signals recorded from multiple electrodes in visual cortex of the monkey during different behavioral states. We asked whether such signals demonstrated coherent fluctuations over time-scales of seconds and minutes, and would thus serve as good candidates for direct comparison with data obtained from functional magnetic resonance imaging (fMRI). We obtained the following results. (i) The BLP of the local field displayed fluctuations at many time-scales, with particularly large amplitude at very low frequencies (<0.1 Hz). (ii) These fluctuations exhibited high coherence between electrode pairs, particularly for BLP signals derived from the gamma frequency range. (iii) Coherence in the BLP, unlike that in the raw LFP, did not fall off sharply as a function of cortical distance. (iv) The structure and coherence of BLP changes were highly similar under distinctly different behavioral states. These results demonstrate the existence of widespread coherent activity fluctuations in the brain of the awake monkey over very long time-scales. We propose that such signals may make a significant contribution to the high variability observed in the time course of physiological signals, including those measured with functional imaging techniques. The results are discussed in the context of combined fMRI/electrophysiological recordings. PMID- 12631573 TI - Reappraisal of serum starvation, the restriction point, G0, and G1 phase arrest points. AB - The restriction point in the G1 phase of the mammalian cell cycle is the oldest, best-known, and widely accepted control point regulating division cycle in mammalian cells. Origins of the restriction point and its subsequent history are reanalyzed here. The initial proposal of the restriction point has an alternative explanation, which is that cells arrested with a G1 phase amount of DNA can arise from the inhibition of a process or processes occurring throughout the cell cycle and are not restricted to any particular phase of the cell cycle or specifically related to any event in the G1 phase of the cell cycle. The initial evidence and subsequent analyses require reexamination. It is proposed that the arrest of cells with a particular DNA content equivalent to that in cells in the G1 phase of the division cycle does not mean there is any particular G1 phase control point. PMID- 12631574 TI - Glial neuronal signaling in the central nervous system. AB - Glial cells are known to interact extensively with neuronal elements in the brain, influencing their activity. Astrocytes associated with synapses integrate neuronal inputs and release transmitters that modulate synaptic sensitivity. Glial cells participate in formation and rebuilding of synapses and play a prominent role in protection and repair of nervous tissue after damage. For glial cells to take an active part in plastic alterations under physiological conditions and pathological disturbances, extensive specific signaling, both within single cells and between cells, is required. In recent years, intensive research has led to our first insight into this signaling. We know there are active connections between astrocytes in the form of networks promoting Ca2+ and ATP signaling; we also know there is intense signaling between astrocytes, microglia, oligodendrocytes, and neurons, with an array of molecules acting as signaling substances. The cells must be functionally integrated to facilitate the enormous dynamics of and capacity for reconstruction within the nervous system. In this paper, we summarize some basic data on glial neuronal signaling to provide insight into synaptic modulation and reconstruction in physiology and protection and repair after damage. PMID- 12631575 TI - One-thousand-and-one substrates of protein kinase CK2? AB - CK2 (formerly termed "casein kinase 2") is a ubiquitous, highly pleiotropic and constitutively active Ser/Thr protein kinase whose implication in neoplasia, cell survival, and virus infection is supported by an increasing number of arguments. Here an updated inventory of 307 CK2 protein substrates is presented. More than one-third of these are implicated in gene expression and protein synthesis as being either transcriptional factors (60) or effectors of DNA/RNA structure (50) or translational elements. Also numerous are signaling proteins and proteins of viral origin or essential to virus life cycle. In comparison, only a minority of CK2 targets (a dozen or so) are classical metabolic enzymes. An analysis of 308 sites phosphorylated by CK2 highlights the paramount relevance of negatively charged side chains that are (by far) predominant over any other residues at positions n+3 (the most crucial one), n+1, and n+2. Based on this signature, it is predictable that proteins phosphorylated by CK2 are much more numerous than those identified to date, and it is possible that CK2 alone contributes to the generation of the eukaryotic phosphoproteome more so than any other individual protein kinase. The possibility that CK2 phosphosites play some global role, e.g., by destabilizing alpha helices, counteracting caspase cleavage, and generating adhesive motifs, will be discussed. PMID- 12631576 TI - Disordered osteoclast formation and function in a CD38 (ADP-ribosyl cyclase) deficient mouse establishes an essential role for CD38 in bone resorption. AB - We have evaluated the role of the ADP-ribosyl cyclase, CD38, in bone remodeling, a process by which the skeleton is being renewed constantly through the coordinated activity of osteoclasts and osteoblasts. CD38 catalyzes the cyclization of its substrate, NAD+, to the Ca2+-releasing second messenger, cyclic ADP-ribose (cADPr). We have shown previously that CD38 is expressed both in osteoblasts and osteoclasts. Its activation in the osteoclast triggers Ca2+ release through ryanodine receptors (RyRs), stimulation of interleukin-6 (IL-6), and an inhibition of bone resorption. Here, we have examined the consequences of deleting the CD38 gene in mice on skeletal remodeling. We report that CD38-/- mice displayed a markedly reduced bone mineral density (BMD) at the femur, tibia, and lumbar spine at 3 months and at the lumbar spine at 4 months, with full normalization of the BMD at all sites at 5 months. The osteoporosis at 3 months was accompanied by a reduction in primary spongiosa and increased osteoclast surfaces on histomorphometric analysis. Hematopoetic stem cells isolated ex vivo from CD38-/- mice showed a dramatic approximately fourfold increase in osteoclast formation in response to incubation for 6 days with RANK-L and M-CSF. The osteoclasts so formed in these cultures showed a approximately 2.5-fold increase in resorptive activity compared with wild-type cells. However, when adherent bone marrow stromal cells were allowed to mature into alkaline phosphatase-positive colony-forming units (CFU-Fs), those derived from CD38-/- mice showed a significant reduction in differentiation compared with wild-type cells. Real-time RT-PCR on mRNA isolated from osteoclasts at day 6 showed a significant reduction in IL-6 and IL-6 receptor mRNA, together with significant decreases in the expression of all calcineurin A isoforms, alpha, beta, and gamma. These findings establish a critical role for CD38 in osteoclast formation and bone resorption. We speculate that CD38 functions as a cellular NAD+ "sensor," particularly during periods of active motility and secretion. PMID- 12631577 TI - Computational dissection of tissue contamination for identification of colon cancer-specific expression profiles. AB - Microarray profiles of bulk tumor tissues reflect gene expression corresponding to malignant cells as well as to many different types of contaminating normal cells. In this report, we assess the feasibility of querying baseline multitissue transcriptome databases to dissect disease-specific genes. Using colon cancer as a model tumor, we show that the application of Boolean operators (AND, OR, BUTNOT) for database searches leads to genes with expression patterns of interest. The BUTNOT operator for example allows the assignment of "expression signatures" to normal tissue specimens. These expression signatures were then used to computationally identify contaminating cells within conventionally dissected tissue specimens. The combination of several logic operators together with an expression database based on multiple human tissue specimens can resolve the problem of tissue contamination, revealing novel cancer-specific gene expression. Several markers, previously not known to be colon cancer associated, are provided. PMID- 12631578 TI - Mechanical stress activates the nuclear factor-kappaB pathway in skeletal muscle fibers: a possible role in Duchenne muscular dystrophy. AB - The ex vivo effects of passive mechanical stretch on the activation of nuclear factor-kappaB (NF-kappaB) pathways in skeletal muscles from normal and mdx mouse, a model of Duchenne muscular dystrophy (DMD), were investigated. The NF kappaB/DNA binding activity of the diaphragm muscle was increased by the application of axial mechanical stretch in a time-dependent manner. The increased activation of NF-kappaB was associated with a concomitant increase in I-kappaB (IkappaB) kinase activity and the degradation of IkappaBalpha protein. Pretreatment of the muscles with nifedipine (a Ca2+ channel blocker) and gadolinium(III) chloride (a stretch-activated channel blocker) did not alter the level of activation of NF-kappaB, ruling out involvement of Ca2+ influx through these channels. Furthermore, N-acetyl cysteine, a free radical inhibitor, blocked the mechanical stretch-induced NF-kappaB activation, suggesting the involvement of free radicals. Compared with normal diaphragm, the basal level of NF-kappaB activity was higher in muscles from mdx mice, and it was further enhanced in mechanically stretched muscles. Furthermore, activation of NF-kappaB and increased expression of inflammatory cytokines IL-1beta and tumor necrosis factor alpha in the mdx mouse precede the onset of muscular dystrophy. Our results show that mechanical stretch activates the classical NF-kappaB pathway and this pathway could be predominately active in DMD. PMID- 12631579 TI - Direct visualization of a stratified epithelium reveals that wounds heal by unified sliding of cell sheets. AB - Observing cells in their original niche is a key link between the information gleaned from planar culture and in vivo physiology and pathology. A new approach combining the transparency of the cornea, Hoffman modulation optics, and digital imaging allowed movements of individual corneal cells to be viewed directly in situ. 3-Dimensional time-lapse movies imaging unstained cells within the stratified corneal epithelium during wound healing were made. Tracking cell movements dynamically provided a definitive answer to the long-standing question: does a stratified epithelium heal by "sliding" of cell sheets as a coherent unit or do individual cells "leap frog" each other at the wound margin? A wound in the corneal epithelium healed primarily by sliding of the whole epithelium, with approximately 95% of cells moving with similar speed and trajectories and with little change in their relative position. Only 5% of cells changed layers, with equal proportions moving up or down. Epithelial healing in situ occurred in three phases: a latency, migration, and reconstruction phase. This model provides a unique system to study the behaviors of individual cells in their original niche. It shows that cells slide into a wound as a unified unit to heal a stratified epithelium. PMID- 12631580 TI - Caspase-dependent cleavage of myosin light chain kinase (MLCK) is involved in TNF alpha-mediated bovine pulmonary endothelial cell apoptosis. AB - Cytoskeletal proteins are key participants in the cellular progression to apoptosis. Our previous work demonstrated the critical dependence of actomyosin rearrangement and MLC phosphorylation in TNF-alpha-induced endothelial cell apoptosis. As these events reflect the activation of the multifunctional endothelial cell (EC) MLCK isoform, we assessed the direct role of EC MLCK in the regulation of TNF-alpha-induced apoptosis. Bovine pulmonary artery endothelial cells expressing either an adenovirus encoding antisense MLCK cDNA (Ad.GFP-AS MLCK) or a dominant/negative EC MLCK construct (EC MLCK-ATPdel) resulted in marked reductions in MLCK activity and TNF-alpha-mediated apoptosis. In contrast, a constitutively active EC MLCK lacking the carboxyl-terminal autoinhibitory domains (EC MLCK-1745) markedly enhanced the apoptotic response to TNF-alpha. Immunostaining in GFP-EC MLCK-expressing cells revealed colocalization of caspase 8 and EC MLCK along actin stress fibers after TNF-alpha. TNF-alpha induced the caspase-dependent cleavage of EC MLCK-1745 in transfected endothelial cells, which was confirmed by mass spectroscopy with in vitro cleavage by caspase 3 at LKKD (D1703). The resulting MLCK fragments displayed significant calmodulin independent kinase activity. These studies convincingly demonstrate that novel interactions between the apoptotic machinery and EC MLCK exist that regulate the endothelial contractile apparatus in TNF-alpha-induced apoptosis. PMID- 12631581 TI - Nitric oxide regulates the polyol pathway of glucose metabolism in vascular smooth muscle cells. AB - Increased reduction of glucose via the polyol pathway enzyme aldose reductase (AR) has been linked to the development of secondary diabetic complications. Because AR is a redox-sensitive protein, which in vitro is readily modified by NO donors, we tested the hypothesis that NO may be a physiological regulator of AR. We found that administration of the NO synthase (NOS) inhibitor N(G)-nitro-L arginine methyl ester (L-NAME) increased sorbitol accumulation in the aorta of nondiabetic and diabetic rats, whereas treatment with L-arginine (a precursor of NO) or nitroglycerine patches prevented sorbitol accumulation. When incubated ex vivo with high glucose, sorbitol accumulation was increased by L-NAME and prevented by L-arginine in strips of aorta from rats or wild-type, but not eNOS deficient, mice. Exposure to NO donors also inhibited AR and prevented sorbitol accumulation in rat aortic vascular smooth muscle cells (VSMC) in culture. The NO donors also increased the incorporation of radioactivity in the AR protein immunoprecipitated from VSMC in which the glutathione pool was prelabeled with [35S]-cysteine. Based on these observations, we suggest that NO regulates the vascular synthesis of polyols by S-thiolating AR; therefore, increasing NO synthesis or bioavailability may be useful in preventing diabetes-induced changes in the polyol pathway. PMID- 12631582 TI - Endocrine regulation of mitochondrial activity: involvement of truncated RXRalpha and c-Erb Aalpha1 proteins. AB - The importance of mitochondrial activity has recently been extended to the regulation of developmental processes. Numerous pathologies associated with organelle's dysfunctions emphasize their physiological importance. However, regulation of mitochondrial genome transcription, a key element for organelle's function, remains poorly understood. After characterization in the organelle of a truncated form of the triiodothyronine nuclear receptor (p43), a T3-dependent transcription factor of the mitochondrial genome, our purpose was to search for other mitochondrial receptors involved in the regulation of organelle transcription. We show that a 44 kDa protein related to RXRalpha (mt-RXR), another nuclear receptor, is located in the mitochondrial matrix. We found that mt-RXR is produced after cytosolic or intramitochondrial enzymatic cleavage of the RXRalpha nuclear receptor. After mitochondrial import and binding to specific sequences of the organelle genome, mt-RXR induces a ligand-dependent increase in mitochondrial RNA levels. mt-RXR physically interacts with p43 and acts alone or through a heterodimerical complex activated by 9-cis-retinoic acid and T3 to increase RNA levels. These data indicate that hormonal regulation of mitochondrial transcription occurs through pathways similar to those that take place in the nucleus and open a new way to better understand hormone and vitamin action at the cellular level. PMID- 12631583 TI - Signaling of hypoxia-induced autonomous proliferation of endothelial cells. AB - Endothelial cells exhibit an autonomous proliferative response to hypoxia, independent of paracrine effectors. In cultured endothelial cells of porcine aorta, we analyzed the signaling and compared hypoxia with mitochondrial inhibition by rotenone. Particularly, roles of the mitogen-activated protein kinase (MAPK) kinase (MEK)/MAPK pathway and cytosolic Ca2+ were studied. Hypoxia resulted in increased proliferation by 65+/-2%. Hypoxia induced transient activation of p42 MAPK (phosphorylation rose from 11+/-5 to 51+/-7%), followed by translocation of p42 MAPK into the nucleus. The proliferative response was diminished after inhibition of the MEK/MAPK pathway by PD 98059 (20 microM) or UO 126 (10 microM) but not sensitive to 8-phenyl-theophillin (10 microM), an adenosine receptor blocker, nor to a neutralizing antibody for vascular endothelial growth factor (VEGF). Inhibition of intracellular Ca2+ release, capacitive Ca2+ influx, or removal of extracellular Ca2+ prevented hypoxic Ca2+ overload and the proliferative response. Suppression of cytosolic Ca2+ rise did not interfere with activation of p42 MAPK but abolished its nuclear translocation. Effects of hypoxia were mimicked by rotenone (10 microM. Transient hypoxic inhibition of mitochondria induces a proliferative endothelial response mediated through Ca2+-independent activation and Ca2+-dependent nuclear translocation of p42 MAPK. This proliferative response is independent of adenosine or VEGF. PMID- 12631585 TI - Activation of microglia by human neuromelanin is NF-kappaB dependent and involves p38 mitogen-activated protein kinase: implications for Parkinson's disease. AB - It has been suggested that microglial inflammation augments the progression of Parkinson's disease (PD). However, endogenous factors initiating microglial activation are largely unknown. We therefore investigated the effects of human neuromelanin (NM) on the release of neurotoxic mediators and the underlying signaling pathways from rat microglia in vitro. The addition of NM to microglial cultures induced positive chemotactic effects, activated the proinflammatory transcription factor nuclear factor kappaB (NF-kappaB) via phosphorylation and degradation of the inhibitor protein kappaB (IkappaB), and led to an up regulation of tumor necrosis factor alpha, interleukin-6, and nitric oxide. The impairment of NF-kappaB function by the IkappaB kinase inhibitor sulfasalazine was paralleled by a decline in neurotoxic mediators. NM also activated p38 mitogen-activated protein kinase (MAPK), the inhibition of this pathway by SB203580 diminished phosphorylation of the transactivation domain of the p65 subunit of NF-kappaB. These findings demonstrate a crucial role of NM in the pathogenesis of PD by augmentation of microglial activation, leading to a vicious cycle of neuronal death, exposure of additional neuromelanin, and chronification of inflammation. The antagonization of microglial activation by a pharmacological intervention targeting microglial NF-kappaB or p38 MAPK could point to additional venues in the treatment of PD. PMID- 12631587 TI - Experimental Biology 2003: April 11-15, San Diego, CA. PMID- 12631584 TI - In vivo selective and distant killing of cancer cells using adenovirus-mediated decorin gene transfer. AB - Decorin is a well-known, ubiquitous proteoglycan that is a normal component of the ECM. Upon transgenic expression of decorin, tumor cells with diverse histogenetic background overexpress p21WAF1, a potent inhibitor of cyclin dependent kinase activity, become arrested in G1, and fail to generate tumors in immunocompromised animals. Because decorin is a secreted protein, it has been recently suggested that decorin could act as an autocrine and paracrine regulator of tumor growth. Here, we demonstrate that adenovirus (Ad)-mediated transfer and expression of human decorin cDNA induced in vivo apoptosis of xenograft tumor cells in nude mice. This oncolytic activity was observed when the Ad vector encoding the decorin cDNA was injected intratumorally (i.t.) or i.v. Importantly, i.t. injection of the decorin Ad vector led to growth inhibition of the injected tumor associated with similar growth inhibition of a distant contralateral tumor, demonstrating a distant decorin antitumoral effect. Immunochemistry against human decorin and decorin quantitation in tumors confirmed that decorin migrated to the tumor distant site. Furthermore, decorin effect was specific to tumor cells, because neither apoptosis nor growth inhibition were observed in nontumoral human cells such as hepatocytes, endothelial cells, and fibroblasts, despite p21 overexpression. PMID- 12631586 TI - Overexpression of wild-type Galpha(i)-2 suppresses beta-adrenergic signaling in cardiac myocytes. AB - The role of Galpha(i)-2 overexpression in desensitization of beta-adrenergic signaling in heart failure is controversial. An adenovirus-based approach was used to investigate whether overexpression of Galpha(i)-2 impairs beta-adrenergic stimulation of adenylyl cyclase (AC) activity and cAMP levels in neonatal rat cardiac myocytes (NRCM) and cell shortening of adult rat ventricular myocytes (ARVM). Infection of NRCM with Ad5Galpha(i)-2 increased Galpha(i)-2 by 50-600% in a virus dose-dependent manner. Overexpression was paralleled by suppression of GTP- and isoprenaline-stimulated AC by 10-72% (P<0.001) in a PTX-sensitive manner. Isoprenaline-stimulated shortening of Ad5Galpha(i)-2-infected ARVM was attenuated by 34% (P<0.01). Ad5Galpha(i)-2/GFP (Galpha(i)-2, green fluorescent protein; bicistronic) was constructed to monitor transfection homogeneity and target Galpha(i)-2 overexpression to levels found in heart failure. At Galpha(i) 2 levels of 93% above control, isoprenaline-stimulated AC activity and cAMP levels were reduced by 17% and 40% (P<0.02), respectively. Beta1- and beta2 adrenergic stimulation was reduced similarly. Our results suggest that (a) the Galpha(i)-2 system exhibits tonic inhibition of stimulated AC in cardiac myocytes, (b) Galpha(i)-2-mediated inhibition is concentration-dependent and occurs at Galpha(i)-2 levels seen in heart failure, and (c) Galpha(i)-2-mediated inhibition affects both beta1- and beta2-adrenergic stimulation of AC. The data argue for an important, independent role of the Galpha(i)-2 increase in heart failure. PMID- 12631588 TI - A model to select regimens for phase III trials for patients with advanced-stage non-small cell lung cancer. AB - PURPOSE: Historical data from pilot, Phase II, and Phase III studies for patients with advanced-stage non-small cell lung cancer (NSCLC) were used to evaluate a statistical model developed to provide assistance in selecting regimens from pilot studies for subsequent use in larger Phase III randomized studies. EXPERIMENTAL DESIGN: Information from 33 Phase III trials for patients with advanced-stage NSCLC performed from 1973 and 1994 in the United States and Canada was collected. The data from antecedent pilot or Phase II and subsequent Phase III trials were analyzed using a predictive statistical model. This model uses the number of patients in the pilot/Phase II study, the median survival of patients in the pilot, and the number of deaths observed, to estimate the statistical likelihood that the pilot regimen will be shown superior to standard therapy in a subsequent Phase III trial. RESULTS: Ten pilot/Phase II studies were identified that preceded eleven subsequent Phase III studies. The three pilot regimens associated with Phase III trials, revealing statistically significant longer survival, had an expected power of 0.69, 0.85, and 0.94 respectively. The regimens from the seven other pilot studies for which the median power expected was 0.38 (range, 0.07-0.80) showed no difference when compared with standard treatment in a Phase III trial. CONCLUSION: The use of the expected power model provides an important enhancement to the screening of new therapies. Regimens with an expected power of >0.55 may be good candidates for testing in Phase III trials. PMID- 12631589 TI - Amplification of erbB2 and erbB2 expression are superior to estrogen receptor status as risk factors for distant recurrence in pT1N0M0 breast cancer: a nationwide population-based study. AB - PURPOSE: To assess the relative importance of 10 prognostic factors in pT1N0M0 breast cancer (< or =2 cm in diameter, node negative). EXPERIMENTAL DESIGN: Women diagnosed with breast cancer in Finland from 1991 to 1992 were identified from the files of the Finnish Cancer Registry, and individual clinicopathological data were collected from the hospital case records of women living in five regions comprising about one-half of the Finnish population. Of the women with minimum required information available (n = 2842), 852 had unilateral pT(1)N(0)M(0) cancer. The median follow-up time was 9.5 years, and only 5% had received systemic adjuvant therapy. Estrogen receptor (ER), progesterone receptor, erbB2, p53, and Ki-67 expression was determined from tumor tissue microarrays using immunohistochemistry, and the erbB2 (HER-2) amplification status was determined using chromogenic in situ hybridization. RESULTS: Primary tumor size < or =5 mm and histological grade 1 were associated with 100 and 95% (95% confidence interval, 92-98%) 9-year distant disease-free survival, respectively, whereas strong erbB2 expression or the presence of >20% Ki-67-positive cells was associated with >20% risk. ER and progesterone receptor values obtained from the hospital case records or tumor microarrays showed weaker association with outcome than the erbB2 status. Small (< or =10 mm) erbB2-negative cancers were associated with >90% 9-year distant disease-free survival, irrespective of histological grade. CONCLUSIONS: Prognosis of pT(1)N(0)M(0) breast cancer is generally well defined by the histological grade and primary tumor size. The erbB2 status was superior to ER as a prognostic factor in these tumors. PMID- 12631590 TI - Identification and validation of genes involved in the pathogenesis of colorectal cancer using cDNA microarrays and RNA interference. AB - PURPOSE: The purpose of this study was to profile gene expression changes in colorectal tumors to identify new targets and strategies for the management of this disease. EXPERIMENTAL DESIGN: cDNA microarray analysis was used to detect differences in gene expression between normal tissue and colon tumors and polyps isolated from 20 patients. To identify genes that are important in regulating the growth properties of colorectal cancer, RNA interference (RNAi) was used to disrupt expression of several of the overexpressed genes in a colon tumor cell line, HCT116, which showed similar patterns of gene expression as many of the patient tumors. RESULTS: Expression changes of > or =2-fold in approximately one third of the patients were consistently observed for 2632 of a total of 9592 genes (574 up-regulated genes and 2058 down-regulated genes). Subsequent analysis of 13 genes by quantitative real-time PCR confirmed the reliability of this analysis. RNAi-mediated disruption of the expression of one of these genes, survivin, a potent inhibitor of apoptosis, severely reduced tumor growth both in vitro and in an in vivo xenograft model. CONCLUSIONS: The combined use of microarray analysis and RNAi provides an excellent system to define the role of specific genes that are up-regulated in cancer lead to the increased in vitro and in vivo growth of colon tumors. PMID- 12631591 TI - MAGE-6 encodes HLA-DRbeta1*0401-presented epitopes recognized by CD4+ T cells from patients with melanoma or renal cell carcinoma. AB - CD4+ T cells modulate the magnitude and durability of CTL responses in vivo and may serve as potent effector cells within the tumor microenvironment. The current study was undertaken to define novel epitopes from the broadly expressed tumor antigen MAGE-6 that are recognized by CD4+ T cells. We have combined the use of a HLA-DR4/peptide binding algorithm with the IFN-gamma enzyme-linked immunospot assay to identify four nonoverlapping sequences derived from the MAGE-6 protein that served as CD4+ T-cell epitopes in HLA-DR4+ donors. Strikingly, patients with active melanoma or renal cell carcinoma failed to secrete IFN-gamma in response to MAGE-6-derived epitopes, whereas both normal donors and cancer patients with no current evidence of disease were responsive, particularly after short-term in vitro stimulations with peptide-pulsed dendritic cells. Importantly, peptide specific CD4+ T cells also recognized HLA-DRbeta1*0401+ tumor cells that constitutively expressed the MAGE-6 protein and autologous HLA-DRbeta1*0401+ dendritic cells transfected with MAGE-6 cDNA-elicited CD4+ T cells that reacted against individual peptide epitopes in vitro. These data suggest that MAGE-6 derived epitopes could serve as useful vaccine candidate components and may provide an immune-monitoring index of clinically important Th1-type immunity in patients with renal cell carcinoma or melanoma. PMID- 12631592 TI - Automated quantification of apoptosis after neoadjuvant chemotherapy for breast cancer: early assessment predicts clinical response. AB - PURPOSE: A large body of evidence implicates apoptosis in the effects of cancer chemotherapeutic agents on tumor cells in vitro and tumor xenografts in vivo, but the predictive value of apoptosis as an early marker for clinical response in cancer patients remains unclear. EXPERIMENTAL DESIGN: We developed an automated, laser scanning cytometer-based method to quantify the percentage of tumor cells containing DNA fragmentation characteristic of apoptosis in tumor sections. We measured levels of apoptosis in a panel of 15 matched, 18-gauge core breast cancer biopsies obtained before and 48 h after neoadjuvant therapy with docetaxel plus doxorubicin or paclitaxel as part of two prospective clinical trials. RESULTS: The results revealed a strongly significant (P = 0.0023) association between chemotherapy-induced apoptosis and pathological response. CONCLUSIONS: If the results can be validated in a larger patient cohort, the method could be used to "tailor" therapy to optimize benefit in a patient-specific fashion. PMID- 12631593 TI - Tumor cyclooxygenase 2-dependent suppression of dendritic cell function. AB - Dendritic cells (DCs) serve as professional antigen-presenting cells and are pivotal in the host immune response to tumor antigens. To define the pathways limiting DC function in the tumor microenvironment, we assessed the impact of tumor cyclooxygenase (COX)-2 expression on DC activities. Bone marrow-derived DCs were cultured in either tumor supernatant (TSN) or TSN from COX-2-inhibited tumors. After culture, DCs were pulsed with tumor-specific peptides, and their ability to generate antitumor immune responses was assessed following injection into established murine lung cancer. In vitro, DC phenotype, alloreactivity, antigen processing and presentation, and interleukin (IL)-10 and IL-12 secretion were evaluated. DCs cultured in TSN failed to generate antitumor immune responses and caused immunosuppressive effects that correlated with enhanced tumor growth. However, genetic or pharmacological inhibition of tumor COX-2 expression restored DC function and effective antitumor immune responses. Functional analyses indicated that TSN causes a decrement in DC capacity to (a) process and present antigens, (b) induce alloreactivity, and (c) secrete IL-12. Whereas TSN DCs showed a significant reduction in cell surface expression of CD11c, DEC-205, MHC class I antigen, MHC class II antigen, CD80, and CD86 as well as a reduction in the transporter-associated proteins, transporter associated with antigen processing 1 and 2, the changes in phenotype and function were not evident when DCs were cultured in supernatant from COX-2-inhibited tumors. We conclude that inhibition of tumor COX-2 expression or activity can prevent tumor-induced suppression of DC activities. PMID- 12631594 TI - Phase I/II clinical study of pulsed paclitaxel radiosensitization for thoracic malignancy: a therapeutic approach on the basis of preclinical research of human cancer cell lines. AB - PURPOSE: A Phase I/II clinical study using pulsed low-dose paclitaxel and radiation for thoracic malignancy was conducted. The study was based on preclinical research of the effects of paclitaxel on apoptosis and the cell cycle in human cancer cell lines. EXPERIMENTAL DESIGN: Three human epithelial cancer cell lines were investigated for preclinical study. Cells were analyzed for apoptosis and cell cycle characteristics after paclitaxel treatment. The Phase I/II clinical trial for non-small cell lung cancer used pulsed low-dose paclitaxel three times/week with the starting dose of 15 mg/m(2). Daily thoracic radiotherapy was delivered in 1.8 Gy/fraction to 60-65 Gy for gross disease and to 45-58 Gy for microscopic disease. Timing of radiotherapy was delayed to allow for a minimum of 4 h for cell cycle progression. RESULTS: Forty-one patients have enrolled and 33 completed treatments. Seventeen patients completed the Phase I study, with an average primary tumor shrinkage of 83 +/- 8% (95% confidence interval). Tumor response rate was 100% for the Phase I study. Overall local control was 98%, and the survival rate was 46% at 1 year, 33% at 2 years, and 18% at 3 years. Toxicity was low with 3 of 18 patients having grade 3 pneumonitis and 3 of 18 patients having grade 3 esophagitis. There was no grade 4 pneumonitis, esophagitis, or hematological toxicity. CONCLUSIONS: Pulsed low-dose paclitaxel radiosensitization for non-small cell lung cancer resulted in a superior local control rate and comparable survival rate when compared with chemoradiation regimens using systemic dose chemotherapy. The regimen is associated with low toxicity and deserves additional investigation, particularly in patients with poor performance or older age, who cannot tolerate standard chemoradiation regimens. PMID- 12631595 TI - Development of Varicella-Zoster virus infection in patients with chronic myelogenous leukemia treated with imatinib mesylate. AB - PURPOSE: Infection with Varicella-Zoster virus (VZV) is an exceptionally rare complication of chronic myelogenous leukemia (CML) without stem cell transplantation. We report 16 patients with CML who developed VZV infection during imatinib mesylate therapy. PATIENTS AND METHODS: From July 1998 until February 2002, 771 patients were included in 11 imatinib mesylate studies for all CML phases in the Departments of Leukemia and Bioimmunotherapy at The University of Texas M. D. Anderson Cancer Center. Sixteen patients developed VZV infection. Charts and follow-up information of were reviewed and analyzed. RESULTS: Sixteen patients (2%) developed a VZV infection [15 episodes of herpes zoster (HZ), 1 varicella]. The baseline characteristics of the 16 patients with infection do not differ significantly from those who did not develop VZV infection, except for time from diagnosis of CML to imatinib (median: 55 versus 25 months, P = 0.0056) and the number of prior therapies (3 versus 1, P < 0.001). All patients received therapy with antiviral agents with good response. Six patients developed postherpetic neuralgia. CONCLUSIONS: Our results suggest that imatinib therapy in CML is associated with low incidence of HZ infection. VZV infection is more frequent with longer duration of CML disease and with prior therapy, does not disseminate, responds well to therapy, and does not mandate a recommendation for HZ prophylaxis in such patients. PMID- 12631596 TI - Phase III randomized study of postradiotherapy chemotherapy with combination alpha-difluoromethylornithine-PCV versus PCV for anaplastic gliomas. AB - PURPOSE: In the current study, we sought to determine whether the addition of DFMO (alpha-difluoromethyl ornithine; eflornithine), an inhibitor of ornithine decarboxylase, to a nitrosourea-based therapy procarbazine, 1-(2-chloroethyl)-3 cyclohexyl-1-nitrosourea, vincristine (PCV) would be more effective as a postirradiation adjuvant therapy for anaplastic gliomas (AG) than PCV alone. PATIENTS AND METHODS: After conventional radiation therapy, 249 AG patients were randomized to receive either DFMO-PCV (125 patients) or PCV alone (124 patients), with survival being the primary endpoint and progression-free survival being an important secondary endpoint. The starting dosage of DFMO was 3 grams/m(2) p.o. q. 8 h for 14 days before and 4 weeks after 1-(2-chloroethyl)-3-cyclohexyl-1 nitrosourea; PCV was administered as described previously (1). Clinical and radiological (gadolinium-enhanced magnetic resonance imaging) follow-ups were nominally at the end of each 6- or 8-week cycle (PCV at 6 weeks; DFMO-PCV at 8 weeks). Laboratory evaluations for hematological and other adverse effects were at 2-week intervals. RESULTS: In the DFMO-PCV arm, there were 114 evaluable patients with 78.1% anaplastic astrocytoma (AA), 3.5% anaplastic oligoastrocytoma (AOA), 14% anaplastic oligodendroglioma (AO), and 4.4% other malignant gliomas. These histological groupings were comparable with those of the 114 patients in the PCV arm: (a) 69.3% AA; (b) 7% AOA; (c) 21.1% AO; and (d) 2.6% malignant gliomas. Although improved survival estimates for the DFMO-PCV treatment group persisted over the course of the study, analysis of survival differences over the entire follow-up period did not yield significance (P = 0.11). However, careful analysis of the corresponding hazard and hazard ratio functions indicated that the real treatment difference was limited to the first 24 months of follow-up (P = 0.02). The median progression-free survival for the two treatment groups, as measured from postradiotherapy registration, was 71.1 months for the DFMO-PCV arm and 37.5 months for the PCV-only arm. Median survival, measured from registration, was 75.8 and 61.1 months, respectively, for the DFMO-PCV and PCV arms. The treatment effect persisted when the AA histology was separated from AO and AOA histologies. This effect persisted even after adjusting for the covariates of age, Karnofsky performance status, and extent of surgery. There was a statistically significant increase in grade 3 adverse events for diarrhea and anemia associated with DFMO-PCV. Grade 3 or 4 adverse events of nausea, ototoxicity, and thrombocytopenia were not significantly increased among groups. CONCLUSIONS: The addition of DFMO to the nitrosourea-based PCV regimen in this Phase III study demonstrated a sustained benefit in survival probabilities for AG patients but not in the corresponding hazard rates. Survival analysis from registration found a DFMO-PCV median survival of 6.3 years (49 of 114 events), whereas that for PCV alone was 5.1 years (55 of 114 events). The hazard function demonstrated a difference over the first 2 years of study (hazard ratio 0.53, P = 0.02) but not after 2 years (hazard ratio 1.06, P = 0.84), supporting the conclusion that DFMO adds to the survival advantage of PCV chemotherapy for AG patients by direct temporal interaction with PCV. PMID- 12631597 TI - Overexpression of oncogenic STK15/BTAK/Aurora A kinase in human pancreatic cancer. AB - PURPOSE: Multiple chromosome abnormalities, including gain of chromosome20q, have been detected frequently in human pancreatic cancers. Overexpression of the STK15/BTAK/Aurora A gene located on chromosome 20q13, which encodes a centrosome associated serine/threonine kinase, has been shown to induce chromosomal instability, leading to aneuploidy and cell transformation in multiple in vitro experimental systems. The purpose of this study was to investigate the expression and copy number alteration of STK15 in pancreatic cancer. EXPERIMENTAL DESIGN: STK15 expression at both the mRNA and protein levels together with the copy number of STK15 gene was measured in nine pancreatic carcinoma cell lines: (a) HPAF-II; (b) Aspc-1; (c) Panc-1; (d) Panc-3; (e) Panc-28; (f) Panc-48; (g) HS766T; (h) MIAPaCa-2; and (i) BxPc3. STK15 protein expression was also examined in normal pancreatic tissues and tumors by Western blotting and immunohistochemistry. RESULTS: STK15 was overexpressed in all of the nine cell lines examined, but gene amplification was infrequent. Western Blot analysis of primary tumor tissues revealed 2-10 times overexpression of STK15 protein compared with normal adjacent tissues from pancreatic cancer patients. Concurrent overexpression of cdc20, an STK15-associated protein, and reduced expression of cdc25, a mitosis-activating protein phosphatase, were detected in the same tumor samples. Elevated STK15 protein expression was detected in 22 of 38 tumor sections (58%) from pancreatic cancer patients. The extent of STK15 expression was not significantly correlated with the size, degree of differentiation, and metastasis status of the tumors. CONCLUSIONS: These results show that STK15 is overexpressed in pancreatic tumors and carcinoma cell lines and suggest that overexpression of STK15 may play a role in pancreatic carcinogenesis. PMID- 12631598 TI - Determinant spreading associated with clinical response in dendritic cell-based immunotherapy for malignant melanoma. AB - PURPOSE: The purpose of this study was to determine the toxicity and immunological effects of three different doses and two routes of administration of autologous dendritic cells (DCs) pulsed with the MART-1(27-35) immunodominant epitope. EXPERIMENTAL DESIGN: Eighteen HLA-A*0201-positive subjects with stage III-IV melanoma received three biweekly i.v. or intradermal injections of ex vivo generated myeloid DCs pulsed with MART-1(27-35) epitope. Repeated blood samples were processed to obtain peripheral blood mononuclear cells for immunological analysis using IFN-gamma ELISPOT, MHC class I tetramer, intracellular cytokine staining, and microcytotoxicity assays. RESULTS: The frequency of MART-1/Melan-A (MART-1) antigen-specific T cells in peripheral blood increased in all dose levels as assessed by ELISPOT and MHC class I tetramer assays, but without a clear dose-response effect. The intradermal route generated stronger MART-1 immunity compared with the i.v. route. MART-1-specific immunity did not correlate with clinical outcome in any of the four immunological assays used. However, analysis of determinant spreading to other melanoma antigens was noted in the only subject with complete response to this single-epitope immunization. CONCLUSIONS: Intradermal immunization with MART-1 peptide-pulsed DCs results in an increase in circulating IFN-gamma-producing, antigen-specific T cells. The frequency of these cells did not correlate with response. In contrast, spreading of immune reactivity to other melanoma antigens was only evident in a subject with a complete response, suggesting that determinant spreading may be an important factor of clinical response to this form of immunotherapy. PMID- 12631599 TI - Interethnic difference in the allelic distribution of human epidermal growth factor receptor intron 1 polymorphism. AB - PURPOSE: Epidermal growth factor receptor (EGFR) plays a critical role in signal transduction and is a target for a novel class of anticancer agents that aim to inhibit EGFR-mediated cancer cell growth. Previous studies have demonstrated a dinucleotide (CA)(n) repeat polymorphism in intron 1 of EGFR, ranging from 14 to 21 repeats, that has been suggested to regulate EGFR expression. The longer allele with 21 repeats showed an 80% reduction of gene expression compared with the shorter allele with 16 repeats. Therefore, the evaluation of the allelic distribution of this polymorphism in populations of various ethnic origins will be crucial to understand the interindividual variability in EGFR expression. EXPERIMENTAL DESIGN: We evaluated the influence of ethnicity on this polymorphism by genotyping individuals of Caucasian (n = 183), African-American (n = 84), and Asian (n = 66) background. RESULTS: The frequency of one of the longer alleles, allele 20 is significantly higher in Asian individuals (63% compared with 21% in Caucasians, P = 2 x 10(-18)). In confirmation of prior studies, the shorter allele 16 was the most common allele in Caucasians (43%) and African-Americans (42%), but its frequency was significantly lower in Asians (average 17%, P = 10( 7) compared with Caucasians). CONCLUSION: Major interethnic differences in the allelic frequencies of the EGFR intron 1 polymorphism exist. Our results may contribute to a better understanding of the molecular basis underlying ethnic differences in drug response and may be helpful for future strategies of individualized therapy with EGFR inhibitors. PMID- 12631600 TI - Loss of cellular retinol-binding protein 1 gene expression in microdissected human ovarian cancer. AB - PURPOSE: We have previously found that cellular retinol-binding protein 1 (CRBP1),involved in retinol transport and metabolism, is down-regulated in an in vitro rat model of ovarian cancer and in several human ovarian cancer cell lines. The aim of this study was to determine the clinical relevance of this change to human ovarian cancer. EXPERIMENTAL DESIGN: A cohort of 48 frozen human serous ovarian carcinomas was evaluated for CRBP1 gene expression. Malignant ovarian epithelial cells were selectively procured by laser capture microdissection, and their CRBP1 expression was determined by real-time PCR. Immunohistochemistry for CRBP1 was performed on paraffin sections of ovarian tumors using polyclonal affinity-purified rabbit anti-CRBP1 antibody. RESULTS: In 35% of ovarian cancer patient samples, there was no detectable CRBP1 expression by real-time PCR. The expression of CRBP1 in microdissected serous ovarian carcinomas was not related to either tumor stage (P = 0.6839) or grade (P = 0.9599). Quantitative PCR results were confirmed by immunohistochemistry using an antibody against CRBP1. CONCLUSIONS: The loss of CRBP1 expression in clinical ovarian tumor specimens is consistent with our previous findings in the rat model and human ovarian cancer cell lines. It appears to be an early event in ovarian carcinogenesis because there was no statistically significant difference in its frequency between tumor stages and grades. Our findings suggest that the loss of CRBP1 expression contributes to the ovarian cancer oncogenesis via altered vitamin A metabolism. PMID- 12631601 TI - Increased activation of CCAAT/enhancer binding protein-beta correlates with the invasiveness of renal cell carcinoma. AB - Positive inflammatory reactions in an aggressive phenotype are typical features of renal cell carcinoma (RCC). Although a high blood level of inflammatory cytokines, such as interleukin-6, interleukin-8, and tumor necrosis factor-alpha, has been observed in these patients, the mechanisms underlying this clinical phenomenon remain to be elucidated. CCAAT/enhancer binding protein (C/EBP) family are transcription factors which play a role in cell differentiation and inflammatory reactions. Among these, C/EBP-beta induces a variety of cytokines and thus may play a role in the pathogenesis of RCC. We studied the activation of C/EBP-beta determined by electrophoretic mobility shift assay in nine RCC cell lines and 44 tissue samples. Six cell lines showed an activation of C/EBP-beta, whereas three cell lines did not, and two of these three had no expression at all of C/EBP-beta protein. Of 44 tissue samples, 12 (27.3%) showed a >200% increase in the activity compared with the corresponding normal kidney tissues. Locally advanced cases had a significantly higher rate of increased C/EBP-beta activity (5 of 8 = 62.5% in advanced cases versus 7 of 36 = 19.4% in localized cases). Especially, all four cases with renal vein invasion had an increased C/EBP-beta activity. These data suggest that the increased activation of C/EBP-beta may contribute to promote tumor invasiveness and render a malignant phenotype of RCC, although it needs to be validated in a larger series. PMID- 12631602 TI - p21 expression predicts outcome in p53-null ovarian carcinoma. AB - PURPOSE: p21 is a direct p53 response gene. Although several studies have correlated p21 and p53 expression, only one has evaluated p21 expression as a function of sequenced p53 gene mutation. We hypothesize that such an analysis may be useful in prognosticating outcome for individuals diagnosed with epithelial ovarian cancer. EXPERIMENTAL DESIGN: DNA from the primary ovarian cancers of 267 patients was studied. p53 mutations were directly sequenced. Two percent or greater nuclear staining with WAF1/CIP1 monoclonal antibody was determined by a hazard ratio analysis to constitute positive p21 expression. RESULTS: Positive p21 nuclear staining occurred more frequently in p53 wild-type ovarian tumors than tumors found to have a p53 mutation (P = 0.001). Positive p21 staining conferred an overall survival advantage (P = 0.02). p21 expression in cancers with p53 missense mutations was not prognostic but did show a strong trend toward significance in the wild-type p53 subset (P = 0.056). Surprisingly, positive p21 staining reflected compromised survival for individuals with p53-null ovarian cancers (P = 0.005). The mean expression level for p21-positive stains in the wild-type group was greater than in null p53 cancers (23 versus 11%; P = 0.001). A Cox multivariable analysis revealed p21 to be a strong independent prognostic factor in p53-null ovarian cancer (P = 0.02). CONCLUSION: p21 expression is closely related to sequenced p53 mutations. This is the first study of positive p21 staining as an independent poor prognostic factor in p53-null ovarian cancer. A dual role for p21 activity, dependent on levels of expression, appears to explain these paradoxical results and is consistent with a complex model for regulation of p21. PMID- 12631603 TI - Detection of hypermethylated RIZ1 gene in primary tumor, mouth, and throat rinsing fluid, nasopharyngeal swab, and peripheral blood of nasopharyngeal carcinoma patient. AB - PURPOSE: The aims of this study are to examine the methylation status of the Fn2 RIZ1 gene in nasopharyngeal (NP) tumors and nasopharyngeal carcinoma (NPC) cell lines and to evaluate the clinical value of methylated RIZ1 gene in body fluid samples of NPC patients. EXPERIMENTAL DESIGN: Methylation status of RIZ1 was evaluated by MSP on CNE-2 and M1 cell lines, 30 tumor biopsies, and their matched body fluid samples, including mouth and throat (M & T) rinsing fluid, NP swabs, plasma, and buffy coat. Normal controls included 8 normal NP biopsies and body fluid samples from 29 healthy volunteers. Sequencing was performed on MSP products from one NP tumor and one M & T rinsing fluid. Transcription of the RIZ1 gene before and after 5-aza-2'-deoxycytidine treatment was examined on CNE-2. RESULTS: The methylated RIZ1 gene was detected in both CNE-2 and M1, 18 (60%) NP tumors, but not in any of the normal controls. Of 30 matched body fluid samples, methylated RIZ1 DNA was found in 11 (37%) NP swabs, 9 (30%) M & T rinsing fluid, 7 (23%) plasma, and 3 (10%) buffy coat samples. Sequencing analysis confirmed all cytosines to uracils conversion, excluding cytosines in CpG dinucleotides in methylated PCR products. Promoter methylation correlated with loss of RIZ1 mRNA expression, and 5-aza-2'-deoxycytidine treatment restored its expression in CNE 2. CONCLUSION: Our results suggest that promoter hypermethylation of the RIZ1 gene is commonly found in NPC. Its detection in body fluid samples of NPC patients but not in normal controls indicates that it is worth to further evaluate its clinical application in assisting screening of NPC and monitoring recurrence after treatment. PMID- 12631604 TI - Twenty-year results of the Naples GUN randomized trial: predictive factors of adjuvant tamoxifen efficacy in early breast cancer. AB - PURPOSE: Tamoxifen (TAM) is increasingly administered to new early breast cancer patients. Because it is not devoid of toxic effects, we studied factors potentially predictive of its efficacy. EXPERIMENTAL DESIGN: From 1978 to 1983, 433 patients were enrolled in the GUN randomized trial: 206 were assigned to TAM versus 227 controls (no-TAM). Premenopausal patients with axillary lymph node involvement (60 TAM versus 65 no-TAM) also received nine CMF cycles. Eight biological markers were retrospectively assayed for most patients: estrogen; progesterone; prolactin receptors (PrlRs); microvessel count (MVC); S-phase fraction; tumor ploidy; epidermal growth factor receptor (EGFR); and HER2. We performed a multivariate test of the TAM/covariate interactions to establish whether these variables predicted for TAM efficacy. Estimates of the TAM effect were expressed as hazard ratio (HR) of death of TAM over no-TAM patients with 95% confidence intervals (95% CIs). RESULTS: At a median follow-up of 15 years, PrlRs, MVC, S-phase fraction, ploidy, and EGFR did not influence TAM efficacy. Differently, HER2 had an overall significant predictive effect: HR = 0.59 (95% CI: 0.40-0.87) in HER2-negative subjects versus HR = 1.09 (95% CI: 0.63-1.87) in HER2-positive subjects (interaction test: P = 0.04). The predictive effect of HER2 was also evident in the subgroup of patients with steroid receptor-positive tumors (HER2 positive: HR = 1.33, 95% CI: 0.70-2.51; HER2 negative: HR = 0.73, 95% CI: 0.47-1.14). CONCLUSIONS: With the statistical power of the present randomized trial, S-phase, ploidy, EGFR, PrlR, and MVC do not seem to predict for TAM efficacy. Conversely, our data support the hypothesis that tumors overexpressing HER2 might not benefit from adjuvant TAM. PMID- 12631605 TI - Quantitative analysis of tumor-derived methylated p16INK4a sequences in plasma, serum, and blood cells of hepatocellular carcinoma patients. AB - PURPOSE AND EXPERIMENTAL DESIGN: Using real-time quantitative methylation specific PCR (RTQ-MSP), we quantified methylated p16INK4a sequences and determined the fractional concentrations of circulating tumor DNA in plasma, serum, and peripheral blood cells collected preoperatively, intraoperatively, and postoperatively from 49 patients with hepatocellular carcinoma (HCC). RESULTS: RTQ-MSP was sufficiently sensitive to detect down to 10 genome-equivalents of methylated p16INK4a sequences. Quantitative MSP data were expressed in terms of the methylation index, which was the percentage of bisulfite converted unmethylated and methylated p16INK4a sequences that consisted of methylated p16INK4a sequences. Quantities of methylated p16INK4a sequences were detected in peripheral circulation of 80% (23 of 29) of HCC patients. No significant difference was seen in the detectability and concentrations of methylated p16INK4a sequences (range: 10-4046 genome-equivalents/ml) between preoperative plasma and serum samples from HCC patients. Preoperatively, the p16INK4a methylation indices ranged from 0.2 to 100% and from 0.012 to 0.075% in the patients' plasma and buffy coat samples, respectively. After surgical resection, the median p16INK4a methylation indices in plasma and buffy coat concordantly decreased 12- and 15-fold, respectively. These results demonstrated the clinical usefulness and effectiveness of peripheral blood RTQ-MSP for detecting and monitoring HCC after treatment. Furthermore, none of the intraoperative plasma samples and only two of the intraoperative buffy coat samples were p16INK4a methylation positive. CONCLUSIONS: Quantification of epigenetic changes in peripheral blood by RTQ-MSP is useful for the detection and monitoring of HCC. PMID- 12631606 TI - A cancer-prone case with a background of methylation of p16 tumor suppressor gene. AB - PURPOSE AND EXPERIMENTAL DESIGN: To date, the presence of p16 gene promoter methylation associated with loss of protein expression has been demonstrated frequently in digestive tract cancers. In this study, we tested for the methylation status of p16 promoter in normal tissue specimens using the methylation-specific PCR technique to examine whether p16 methylation already existed in the background of tumors. RESULTS: Aberrant promoter methylation of p16 gene was detected in 1 of 40 esophageal and 1 of 69 gastric and no colorectal epithelium specimens, and these 2 specimens were derived from the same patient. We also found the same methylation change in both tumor and blood cell DNA. CONCLUSION: These results suggested that the p16 gene was inactivated by methylation in normal background cells of this patient and that other additional factors may promote tumor development in his esophageal and gastric tissues. PMID- 12631607 TI - Genomic instability and tumor-specific alterations in oral squamous cell carcinomas assessed by inter-(simple sequence repeat) PCR. AB - PURPOSE: Genomic instability plays a major role in the genesis and progression of tumors, and in the evolution of tumor heterogeneity. To determine the role of genomic instability in the genesis and progression of oral cancer, we assessed the extent of genomic alterations in oral squamous cell carcinomas (OSCCs). EXPERIMENTAL DESIGN: We used the recently developed inter-(simple sequence repeat) PCR technique to quantitate genomic instability using matched tumor and normal OSCC samples (n = 25). The inter-repeat region bands of similar molecular size observed to be altered in more than one case were sequenced and analyzed to identify probable OSSC-associated specific genetic lesions. RESULTS: Of the four base-anchored, dinucleotide repeat-based primers used for the study, the most informative profile in OSCCs was generated by the (CA)(8)RG primer. Measurement of genomic instability index using the (CA)(8)RG primer revealed a high incidence of genomic instability in OSCCs. No significant correlation between the extent of alterations and stage or location of the tumor was observed. Sequencing analysis of the altered bands revealed gains/losses in several chromosomal regions. Of the matched tumor and corresponding normal tissue DNA studied, hitherto unreported losses were seen in 11p15 and 17q25 chromosomal regions. Sequencing of some of the tumor-specific altered regions indicated that they code for regions of UDP GalNAc and hRAD 17 genes, which were lost (deleted) in oral cancer. CONCLUSIONS: Our results indicate that the extent of genomic instability in OSCC is not correlated to the tumor stage or location. For the first time, we have shown that chromosomal alterations detected by inter-(simple sequence repeat) PCR could be correlated to genes associated with cancer development. PMID- 12631608 TI - BCL-6 expression predicts improved survival in patients with primary central nervous system lymphoma. AB - PURPOSE: The purpose of this study was to investigate the histogenetic origin of primary central nervous system lymphoma (PCNSL) with respect to stage of B-cell differentiation and to identify prognostic markers in a cohort of patients with PCNSL treated with i.v. high-dose methotrexate therapy. EXPERIMENTAL DESIGN: This study included 33 patients with PCNSL treated with high-dose i.v. methotrexate at the Massachusetts General Hospital for whom archival tumor tissue was available. All 33 patients tested negative for HIV. The lymphomas were morphologically subclassified according to the Kiel system, as modified in the WHO classification. Immunohistochemistry for the following antigens was performed: BCL-6; BCL-2; MUM1; CD10; vs38c; CD138; CD44; p16; and p53. Fluorescence in situ hybridization and multiplex PCR for CDKN2A/p16 were also performed. RESULTS: There were 17 women and 16 men enrolled, with a median age of 60 years. All tumors were diffuse large B-cell lymphomas. Of the 23 cases that could be subclassified, 22 were centroblastic, and 1 was immunoblastic. Twenty-six of 33 tumors were BCL-6+, 6 of 32 tumors were CD10+, 27 of 29 tumors were BCL-2+, 31 of 32 tumors were MUM1+, 11 of 31 tumors were CD44+, 4 of 33 tumors were vs38c+, and 0 of 32 tumors were CD138+. There were 18 of 32 (56%) complete responses and 8 of 32 (25%) partial responses to methotrexate, whereas 6 of 33 (18%) progressed during treatment. Ten patients died of disease. Expression of BCL-6 was significantly associated with longer overall survival (P = 0.002; median survival, 101 versus 14.7 months, with approximately 95% lower confidence limits of 41.7 and 8.8 months, respectively). CONCLUSIONS: In this group of 33 patients with PCNSL, expression of BCL-6 was significantly associated with longer overall survival. BCL-6 warrants further investigation as a potentially important prognostic marker in this disease. PMID- 12631609 TI - Epidermal growth factor receptor, cyclooxygenase-2, and BAX expression in the primary non-small cell lung cancer and brain metastases. AB - PURPOSE: The purpose is to identify biological markers that predict brain metastasis and treatment outcome in non-small cell lung cancer (NSCLC). EXPERIMENTAL DESIGN: Samples were obtained from the primary tumors, lymph nodes, and brain metastases of 29 patients with NSCLC who had undergone resection of both the pulmonary tumors and the brain lesions. Samples from 29 patients matched for age, sex, and histology whose pulmonary tumors were resected served as controls. Samples were stained with H&E as well as immunohistochemical stains for epidermal growth factor receptor (EGFR), cyclooxygenase 2 (COX-2), and BAX. Comparisons were made between patients with and without brain metastasis. Independent investigators determined the percentage of positive cells. RESULTS: There was positive correlation in expression of all three biomarkers between primary lung tumors and lymph node metastases. Significantly higher levels of EGFR were found in lymph node metastases in the control group (P = 0.0147). COX-2 expression in brain lesions correlated with expression in primary tumors (P = 0.023). BAX levels were lower in poorly differentiated tumors in lymph node metastases in the control group (P = 0.01) and in brain metastases (P = 0.045). Low EGFR expression and high COX-2 expression in lymph node metastasis were associated with poorer treatment outcome. CONCLUSIONS: Expression of EGFR, COX-2, and BAX in primary lung tumors did not differ between patients with brain metastases from NSCLC and those without brain metastases. These three biomarkers cannot be used to predict brain metastasis. Studies of other biomarkers are under way in an effort to predict brain metastasis among patients with NSCLC. PMID- 12631610 TI - Alpha-1-acid glycoprotein as an independent predictor for treatment effects and a prognostic factor of survival in patients with non-small cell lung cancer treated with docetaxel. AB - PURPOSE: To identify predictors of treatment outcome and survival in patients with non-small cell lung cancer (NSCLC) treated with docetaxel. EXPERIMENTAL DESIGN: The data were collected from 180 NSCLC patients enrolled in six docetaxel Phase II studies at a dose of 100 mg/m(2). Clinical end points for this study were safety reported as the first course adverse events requiring dose reduction, and efficacy was measured by response rate and survival. The independent variables included docetaxel dose, individual estimates of clearance, area under the plasma concentration time curve, extent of previous treatment, and covariables related to the patient's demographics, extent of disease, and performance status. The data were analyzed using a logistic regression model for response and severe adverse events and a Cox multivariate regression model for survival. RESULTS: Docetaxel exposure as measured by the area under the plasma concentration time curve was the only significant predictor (P < 0.0001) of severe toxicity during the first course of therapy. Baseline alpha1-acid glycoprotein (AAG) was the only significant predictor of response with an odds ratio of 0.44 for changes in AAG from 1.11 to 1.85 grams/liter (P = 0.0039). Cumulative dose, AAG, and extent of disease were independent predictors of survival (P < 0.005). The median survival varied from 15.6 months for patients with a low AAG (AAG < or = 1.11 grams/liter) to 5.5 months for patients with a high AAG (AAG >/= 1.85 grams/liter). CONCLUSION: AAG appears to be an independent predictor of response and a major objective prognostic factor of survival in patients with NSCLC treated with docetaxel chemotherapy. PMID- 12631611 TI - Response to chemotherapy and expression of the genes encoding the multidrug resistance-associated proteins MRP2, MRP3, MRP4, MRP5, and SMRP in childhood acute myeloid leukemia. AB - PURPOSE: The family of multidrug resistance-associated proteins (MRPs) belongs to the ATP-binding cassette superfamily of transporters, which have the ability to function as outward pumps for chemotherapeutic drugs. Their structure, function, and substrate specificity have been studied intensively, but little is known about their clinical relevance in malignant diseases. EXPERIMENTAL DESIGN: In this study, the expression of the MRP2, MRP3, MRP4, MRP5, and SMRP genes was measured using TaqMan real-time PCR in 53 children with de novo acute myeloid leukemia. Nine patients were also analyzed in relapse. RESULTS: MRP3 gene expression was higher in patients who did not achieve remission (P = 0.023). Expression of MRP2 (P = 0.09) or MRP3 (P = 0.041) was associated with a lower rate of survival, and patients who expressed high levels of both genes had a particularly poor prognosis (P < 0.01). No significant association was found for overall survival or remission rate and the expression of MRP4, MRP5, and SMRP. CONCLUSIONS: This study provides first data on the clinical relevance of five MRPs in acute myeloid leukemia patients. The results strongly suggest that MRP3 and possibly also MRP2 are involved in drug resistance in this disease. Those two proteins therefore represent interesting markers for risk-adapted therapy and possible targets for the development of specific drugs to overcome multidrug resistance. PMID- 12631612 TI - Human epidermal receptor-2 expression in prostate cancer. AB - PURPOSE: Efforts to conclusively establish that human epidermal receptor (HER)-2 overexpression is important to androgen-dependent carcinoma of the prostate (AD CaP) or to progression to androgen independence (AI-CaP) have failed because of variability in tissue procurement, antibodies, immunostaining procedures, and assessment methods. However, because some in vitro and animal model data correlate HER-2 overexpression with progression to androgen independence, trials of agents that target the HER-2 receptor are under way. To clarify human tumor findings, we studied HER-2 expression at the gene (DNA), mRNA, and protein levels in well-characterized CaP specimens. EXPERIMENTAL DESIGN: Fifty AD-CaP and 25 AI CaP specimens from similar numbers of Caucasian and African Americans were immunostained for HER-2 receptor. HER-2 mRNA levels were measured using real-time fluorescence quantitative PCR in patients for whom frozen specimens were available. HER-2 amplification was evaluated using fluorescent in situ hybridization. RESULTS: HER-2 receptor immunostained in 52% of androgen-dependent and one (4%) androgen-independent tumor. HER-2 immunostaining was not related to age, race, serum prostate-specific antigen levels, or pathologic stage and Gleason grade. HER-2 overexpression was not detected in AI-CaP at the mRNA or gene level. Mean HER-2 mRNA expression was higher (P < 0.05) in AD-CaP than AI CaP (22,080 versus 15,496 HER-2 copies). HER-2 was not amplified in any of 20 AD CaP or 19 AI-CaP specimens. CONCLUSIONS: HER-2 protein and message overexpression and HER-2 amplification were not found in AI-CaP. PMID- 12631613 TI - Elevated expression level of survivin protein in soft-tissue sarcomas is a strong independent predictor of survival. AB - PURPOSE: Survivin is a member of the inhibitor-of-apoptosis gene family and is known to be overexpressed in a number of tumor types. The aim of this study was to evaluate the prognostic value of survivin protein expression in tumor tissue extracts in a group of well-characterized soft-tissue sarcoma (STS) patients. EXPERIMENTAL DESIGN: In this investigation, malignant tissue samples from 63 STS patients as well as from a panel of tumor cell lines were investigated, with nonmalignant tissues serving as controls. The survivin protein level was quantified by a novel ELISA and by Western blot analysis. Results obtained by both methods were compared with clinicopathological parameters regarding tumor grade and tumor entity, and they were then correlated to survival in a multivariate Cox regression model. RESULTS: High survivin levels were detected by ELISA and Western blot analysis in tumor tissue extracts and in lysates of tumor cell lines. None or only weak expression of survivin protein was found in nonmalignant cells and tissues. When comparing survivin values obtained by ELISA or Western blot, we found a significant correlation between both methods (P = 0.013, Pearson test). Our findings revealed that, in multivariate Cox regression analyses, survivin levels measured by ELISA and Western blot were significantly associated with tumor-related death in STS patients (P = 0.001, RR = 19.8, and P = 0.004, RR = 5.1, respectively). However, in a direct comparison of both survivin protein detection assays, we found a higher sensitivity and a stronger correlation to prognosis in survivin ELISA as compared with the Western blot assays. Furthermore, a higher tumor grade and more aggressive STS entity showed an elevated survivin protein expression level. CONCLUSION: Altogether, an elevated survivin content in tumor tissue extracts has a significant and independent negative predictive value on the survival-rate of STS patients. This finding corresponds well to data obtained for the mRNA level of survivin, as shown previously (M. Kappler et al., Int. J. Cancer, 95: 360-363, 2001). PMID- 12631614 TI - Expression of mRNAs for telomeric repeat binding factor (TRF)-1 and TRF2 in atypical adenomatous hyperplasia and adenocarcinoma of the lung. AB - PURPOSE AND EXPERIMENTAL DESIGN: It has been suggested that atypical adenomatous hyperplasia (AAH) may be a precursor of peripheral adenocarcinoma of the lung. Telomerase is a ribonucleoprotein enzyme that synthesizes telomeric DNA onto chromosomal ends. Its activity is thought to participate in the development of most human cancers. Telomere-specific DNA-binding proteins, such as telomeric repeat binding factor 1 and telomeric repeat binding factor 2, also control telomere length in a complex interplay with telomerase. Here we investigated the expressions of the mRNAs encoded by the TERF1 and TERF2 genes using in situ hybridization in surgically resected specimens [28 AAHs (11 lesions were interpreted as low-grade AAH, and 17 were interpreted as high-grade AAH) and 40 peripherally located bronchioloalveolar carcinoma (BAC). RESULTS: A clear overexpression of these mRNAs was recognized in low- and high-grade AAH and BAC samples (as compared with normal tissues) using in situ hybridization and these mRNAs were detected in normal AAH and BAC samples using reverse transcription PCR. The expressions of TERF1 and TERF2 mRNA detected by in situ hybridization were scored positive in 36% and 82% of low-grade AAH, 65% and 83% of high-grade AAH, and 88% and 88% of BAC, respectively. Statistically significant differences in TERF1 mRNA expression could be shown between low-grade AAH and BAC and between high-grade AAH and BAC. There was no statistical difference in the positive expressions of TERF2 mRNA among low-grade AAH, high-grade AAH, and BAC. CONCLUSIONS: These results are consistent with (but are not enough to confirm) the idea that high-grade AAH is closely related to BAC. PMID- 12631616 TI - Mouse mammary tumor virus-like gene sequences in breast tumors of Australian and Vietnamese women. AB - PURPOSE: There is considerable evidence that the presence of mouse mammary tumor virus (MMTV)-like gene sequences in human breast cancer is highly associated with human breast carcinoma. Previous studies have found MMTV-like gene sequences in 38% of breast cancer tissue from United States women. The prevalence of these sequences in Australian and Vietnamese women has never been reported. EXPERIMENTAL DESIGN: Using PCR and primers that amplify MMTV-like gene sequences, we tested cancerous and benign breast tissue from Caucasian-Australian, Vietnamese-Australian, and Vietnamese women. RESULTS: MMTV-like gene sequences were amplified in 19 of 45 (42.2%) archival breast cancer biopsy tissues from Caucasian-Australian women, but only 1 of 120 (0.8%) and 0 of 41 breast cancer biopsy tissues from Vietnamese and Vietnamese-Australian women, respectively. The same sequences were found in only 2 of 111 (1.8%) and 0 of 60 normal (benign) breast tissue samples from Australian and Vietnamese women, respectively. CONCLUSIONS: MMTV-like gene sequences are found in only some human populations and are rarely found in normal human breast tissue from all populations, suggesting they are not present in the normal human genome and have been acquired. PMID- 12631615 TI - High incidence of microsatellite instability in colorectal cancer from African Americans. AB - PURPOSE: Colorectal carcinoma (CRC) is the second most common cause of cancer death in the United States, and the rate of CRC is nearly 1.5 times higher in African-Americans (AA) than in Caucasians. Microsatellite instability (MSI) is observed in sporadic CRC reflecting promoter hypermethylation of the DNA mismatch repair gene hMLH1, and anecdotal evidence suggests an increased incidence of MSI among AAs. Additionally, p16 can be inactivated by hypermethylation of the promoter region, abrogating its ability to regulate cell proliferation. The objective of this study is to determine the frequency of MSI and p16 gene methylation in CRC from AA patients. EXPERIMENTAL DESIGN: Experiments were conducted on serially collected archival samples of colon cancer and adjacent normal tissue (n = 22). Five microsatellite markers were used to measure MSI in tumors with direct comparison to normal tissue from the same patient. p16 promoter methylation status was determined by methylation-specific PCR. RESULTS: Ten cancers (45%) demonstrated high MSI (MSI-H), 1 demonstrated low MSI, and the remaining 11 tumors were microsatellite stable. Most of the MSI-H tumors were proximal, well differentiated, and showed high levels of mucin production. Most patients in the MSI-H group were female (70%), whereas most of the microsatellite stable group (81%) were male. Five of the 22 tumors (22%) had methylation of the p16 promoter. CONCLUSION: Data provided here demonstrated that the incidence of MSI-H tumors was 3-fold higher in our study group of AA patients compared with data reported in nonracially selected but serially collected studies. Odds ratio analysis indicates that the chance of female patients having MSI-H was 11.7 times more than male patients (P < 0.03). The reason for this gender difference is unknown. These findings might reflect dietary differences or genetic polymorphisms that may be common in the AA population. Additional investigation in a larger patient population is needed before strong conclusion can be drawn. PMID- 12631617 TI - Expression of p27(Kip1) and c-Jun activation binding protein 1 are inversely correlated in systemic anaplastic large cell lymphoma. AB - PURPOSE: p27(Kip1)(p27) is a universal cyclin-dependent kinase inhibitor that inhibits cell cycle transition from G(1) to S phase and is primarily regulated at the post-transcriptional level via the ubiquitin-proteasome pathway. In vitro data suggest that p27 degradation may be accelerated by the c-Jun activation domain binding protein-1 (JAB1), originally identified as a coactivator of the gene regulatory AP-1 proteins. We assessed p27 and JAB1 in systemic anaplastic large cell lymphoma (ALCL), a group of tumors in which a substantial subset overexpresses anaplastic lymphoma kinase (ALK). EXPERIMENTAL DESIGN: The study included 5 ALK-positive ALCL cell lines, namely Karpas 299, JB-6, SR-786, SU DHL1, and TG-S1, and 66 ALCL tumors (24 ALK positive and 42 ALK negative). The cell lines were analyzed by Western blot methods, and the tumors were assessed immunohistochemically. RESULTS: SU-DHL1 and TG-S1 cells were positive for p27 and negative for JAB1, whereas SR-786 and JB-6 cells were positive for JAB-1 but negative for p27. Karpas 299 expressed p27 at relatively low levels and JAB1 at high levels. Using a 10% cutoff, p27 was positive in 12 of 66 (18.2%) ALCL tumors (5 ALK positive and 7 ALK negative), whereas JAB1 was detected in 47 of 53 (88.7%) tumors (15 ALK positive and 32 ALK negative) assessed. p27 and JAB1 expression were inversely correlated (Spearman r = -0.27, P = 0.03). For 54 ALCL patients with complete follow-up, and in separate analyses of patients with ALK positive or -negative tumors, p27 expression correlated with poorer prognosis. CONCLUSIONS: p27 is absent or expressed at low levels in most ALCL tumors and inversely correlates with JAB1. These findings suggest that JAB1-mediated degradation of p27, allowing cell cycle progression, may play a role in the pathogenesis of ALCL. PMID- 12631618 TI - Significant antitumoral activity of cationic multilamellar liposomes containing human IFN-beta gene against human renal cell carcinoma. AB - PURPOSE: Immunotherapy is the most effective treatment against metastatic renal cell carcinoma (RCC). However, the response rate is approximately 15%. More effective therapy is, therefore, needed for patients with metastatic RCC. We then examined the antitumor effect of cationic multilamellar liposome containing human IFN-beta (huIFN-beta) gene (IAB-1) against RCC. EXPERIMENTAL DESIGN: Concentrations of huIFN-beta protein were measured by ELISA. The cytotoxicity of IAB-1 against human RCC (NC65, ACHN, and freshly isolated RCC cells), prostate and bladder cancer cell lines, and renal proximal tubule endothelial cells (RPTEC5899) was examined by the colorimetric method using tetrazolium salt. Apoptosis was assessed by the acridine-orange staining. For in vivo study, we used NC65 cells inoculated into severe combined immunodeficiency mouse. RESULTS: The RCC cells treated with IAB-1 secreted significant amounts of huIFN-beta protein continuously. Drastic in vitro cytotoxic effect of IAB-1 against RCC was observed. In contrast, treatment with 1000 IU/ml recombinant huIFN-beta protein resulted in weak cytotoxicity. The cytotoxic effect against prostate and bladder cancer cell lines was less than that against RCC. Furthermore, no significant cytotoxicity was observed in RPTEC5899 cells. Apoptosis was observed in the cells treated with IAB-1, but recombinant huIFN-beta failed to induce apoptosis. The size of NC65 tumors transfected with IAB-1 in mice was significantly smaller than that receiving injection of empty liposome or recombinant huIFN-beta protein. CONCLUSION: These findings indicate that IAB-1 may have an antitumor activity against human RCC by inducing apoptosis, suggesting its potential clinical application for gene therapy against RCC. PMID- 12631619 TI - The proteasome inhibitor PS-341 markedly enhances sensitivity of multiple myeloma tumor cells to chemotherapeutic agents. AB - Increased nuclear factor kappaB (NF-kappaB) activity is associated with increased tumor cell survival in multiple myeloma. The function of NF-kappaB is inhibited through binding to its inhibitor, IkappaB. Release of activated NF-kappaB follows proteasome-mediated degradation of IkappaB resulting from phosphorylation of the inhibitor and, finally, conjugation with ubiquitin. We report that myeloma cells have enhanced IkappaBalpha phosphorylation and increased NF-kappaB activity compared with normal hematopoietic cells. The proteasome inhibitor PS-341 blocked nuclear translocation of NF-kappaB, blocked NF-kappaB DNA binding, and demonstrated consistent antitumor activity against chemoresistant and chemosensitive myeloma cells. The sensitivity of chemoresistant myeloma cells to chemotherapeutic agents was markedly increased (100,000-1,000,000-fold) when combined with a noncytotoxic dose of PS-341 without affecting normal hematopoietic cells. Similar effects were observed using a dominant negative super-repressor for IkappaBalpha. Thus, these results suggest that inhibition of NF-kappaB with PS-341 may overcome chemoresistance and allow doses of chemotherapeutic agents to be markedly reduced with antitumor effects without significant toxicity. PMID- 12631620 TI - Mechanisms of proteasome inhibitor PS-341-induced G(2)-M-phase arrest and apoptosis in human non-small cell lung cancer cell lines. AB - PURPOSE: PS-341 is a novel dipeptide boronic acid proteasome inhibitor with in vitro and in vivo antitumor activity that induces mechanisms of apoptosis by unknown mechanisms. EXPERIMENTAL DESIGN: Human non-small cell lung cancer cell lines were used to investigate effects PS-341 on cell proliferation, cell cycle progression, and the induction of apoptosis. RESULTS: PS-341 was 38-360-fold more cytotoxic against H460 cells when compared with the proteasome inhibitors MG-132 and PSI. Differential PS-341 cytotoxic effects were found with respect to P53 function: H322 cells (p53 mutant) were 6-fold less sensitive as compared with H460 cells (p53 wild type); and H358 cells (p53 null) were 1.6-fold more sensitive as compared with H460 cells (p53 wild type). A concentration- and time dependent cell cycle blockade at G(2)-M phase was seen for H460 cells without any direct effects on microtubule polymerization or depolymerization. PS-341 exposure in H460 cells led to stabilization of p53, induction of p21(cip/waf-1) and MDM2 expression, an increase in cyclin B and cyclin A, and the activation of cyclin B and cyclin A kinases. MDM2 induction was found only in H460 cells, whereas in H322 and H358 cells, G(2)-M-phase arrest, p21(cip/waf-1) induction, and an increase in cyclin B1 were found. The commitment of G(2)-M-phase cells to apoptosis was verified by the activation of caspase-3 and cleavage of poly(ADP ribose) polymerase in drug-free medium. CONCLUSIONS: Our data suggest that the PS 341-induced G(2)-M-phase arrest may be associated with the inhibition of degradation of cell cycle regulators and that the up-regulation of p21(cip/waf-1) expression may be via p53-dependent and/or -independent pathways. The resulting disturbance of cell cycle progression leads either to growth inhibition or to the initiation of apoptotic pathways. PMID- 12631622 TI - Raf-1 and Bcl-2 induce distinct and common pathways that contribute to breast cancer drug resistance. AB - Overexpression of Bcl-2 plays a role in the development of drug resistance in leukemia and other apoptosis-prone tumors. Raf isoforms areserine/threonine kinases that act as signal transducers in cascades initiated by many growth factors and mitogens. Raf isoform activation has been linked to drug resistance in leukemia. In this study we investigated effects of Bcl-2 and Raf-1 on doxorubicin-induced growth inhibition of MCF-7 breast cancer cells. In the absence of doxorubicin, overexpression of Bcl-2 or a constitutively active form of Raf-1 in MCF-7 cells did not affect proliferation rate. Overexpression of Bcl 2 increased resistance of MCF-7 cells to doxorubicin in 2-day, 5-day, and 8-week assays. Analysis of doxorubicin sensitivity of individual MCF/Bcl-2 clones showed that doxorubicin resistance was positively correlated with level of Bcl-2 overexpression. Overexpression of constitutively active Raf-1 also increased resistance to doxorubicin. Induction of Raf-1 activity in MCF-7 cells overexpressing Bcl-2 resulted in greater doxorubicin resistance than induction of Raf-1 activity in MCF-7 cells lacking Bcl-2 overexpression. Furthermore, levels of P-glycoprotein mRNA were increased in MCF-7 cells overexpressing a constitutively active Raf-1. MCF-7 cells overexpressing constitutively active Raf 1 were also more resistant to paclitaxel, which, like doxorubicin, is a substrate of P-glycoprotein. These observations suggest both independent and overlapping roles for Raf-1 and Bcl-2 oncogenes in the resistance to growth inhibition by doxorubicin. PMID- 12631621 TI - Increased resistance of tumor cells to hyperthermia mediated by integrin-linked kinase. AB - PURPOSE: Integrin-linked kinase (ILK) is a serine-threonine kinase associated with anchorage-independent growth and tumorigenic transformation. Previous studies indicate that overexpression of ILK is common among several types of tumors, and it is involved in the regulation of tumor cell survival under stress. In this study, we examined the effects of ILK expression on tumor cellular response to hyperthermia. EXPERIMENTAL DESIGN: We used an adenovirus-mediated approach to overexpress the ILK gene in a prostate cancer cell line and examine its effects on heat stress-induced cell death. Clonogenic survival, as well as apoptosis, was evaluated in cells that overexpress ILK. In addition, the ability to form tumors in vivo was examined in syngeneic hosts. Finally, potential molecular mechanisms of ILK-mediated resistance to heat were examined by determining the status of a variety of signal transduction pathways. RESULTS: ILK overexpression made tumor cells significantly more resistant to the cell-killing effects of hyperthermia. This was correlated at the molecular level with the down regulation of hyperthermia-induced activation of stress-activated protein kinase/c-Jun-NH(2)-terminal kinase, p38 mitogen-activated protein kinase activities, and caspase 9. The overexpression of ILK was also shown to induce a more rapid tumor growth in a murine prostate cancer cell line CONCLUSION: ILK plays an important role in tumor growth and tumor response to hyperthermia treatment. PMID- 12631623 TI - Antisense protein kinase A RIalpha acts synergistically with hydroxycamptothecin to inhibit growth and induce apoptosis in human cancer cells: molecular basis for combinatorial therapy. AB - PURPOSE: The increased expression of RIalpha, the regulatory subunit of cyclic AMP (cAMP)-dependent protein kinase type I (PKA-I), has been correlated with cancer cell growth. An antisense oligonucleotide targeting the RIalpha subunit of PKA (antisense RIalpha) induces cell growth arrest, apoptosis, and differentiation in a variety of cancer cell lines in vitro and in tumors in vivo. This study investigated the utility of a combinatorial therapy consisting of the RNA-DNA second-generation RIalpha antisense HYB0165 (Gem231) and the cytotoxic drug hydroxycamptothecin (HCPT), which inhibits topoisomerase I. EXPERIMENTAL DESIGN: LS-174T colon carcinoma and PC3M androgen-insensitive prostate cancer cells were used as experimental models. The antitumor and apoptotic activities of Gem231 and HCPT, singly and in combination, were measured by cell growth assay, synergism quotient, cell morphology, nuclear morphology, levels of PKA R and C subunits, anti- and proapoptotic proteins, and PKA activity ratio. RESULTS: In a synergistic fashion, Gem231 and HCPT induced growth arrest, apoptosis, and changes in cell morphology; down-regulated RIalpha expression; down-regulated Bcl 2 and promoted its hyperphosphorylation; up-regulated the proapoptotic proteins Bax and Bad; and promoted hypophosphorylation of Bad. Antisense Gem231, but not HCPT, increased the PKA activity ratio, which measures the degree of PKA activation. CONCLUSION: The results showed that PKA-I activation by Gem231 and topoisomerase I inhibition by HCPT are responsible at the molecular level for the synergistic effects of tumor cell apoptosis and growth inhibition. These results demonstrated the molecular basis for the use of Gem231 and HCPT as combinatorial therapy to treat human cancer. PMID- 12631624 TI - Using apoptosis for targeted cancer therapy by a new gonadotropin releasing hormone-DNA fragmentation factor 40 chimeric protein. AB - PURPOSE: GnRH-based chimeric proteins have been shown to specifically target and kill adenocarcinomas both in vitro and in vivo. The purpose of this study is to construct a new GnRH-based chimeric protein for the treatment of adenocarcinomas in humans. RESULTS: In this study, we constructed and characterized a new chimeric protein, GnRH-DFF40, composed of a new human killing moiety: the apoptotic DNase-DFF40 (DNA fragmentation factor), known also as caspase-activated DNase (CAD). GnRH-DFF40 exhibits DNase activity in vitro. We found that this chimeric protein can target and kill adenocarcinoma cells. Such death occurs via apoptotic pathways, resulting in an increase in the sub-G(1) population, DNA fragmentation, terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL)-positive cells, and morphology typical of apoptotic cells. These apoptotic events involve the mitochondria because we found cytochrome c depletion and caspase-9 and caspase-3 activation. Preliminary in vivo results showed that treatment of colon adenocarcinoma xenografts in nude mice with the new chimeric protein caused a reduction in tumor weight. CONCLUSIONS: Because GnRH-DFF40 is a whole human-based chimeric protein when applied to humans, the nonspecific toxicity and immunogenicity seen with bacterial/plant-based chimeric proteins should be avoided. Thus, GnRH-DFF40 is a promising candidate for the treatment of adenocarcinomas in humans. PMID- 12631625 TI - Interleukin 10 blocks matrix metalloproteinase-2 and membrane type 1-matrix metalloproteinase synthesis in primary human prostate tumor lines. AB - Insulin-like growth factor (IGF) I has been shown previously to up-regulate matrix metalloproteinase-2 (MMP-2) production, whereas the interleukin (IL) 10/IL 10 receptor axis has been found to down-regulate MMP-2 synthesis in tumor cells. In this paper, we showed that IL-10 activation of the IL-10 receptor blocked MMP 2 and membrane type 1 (MT1) -MMP transcription and protein synthesis in nonimmortalized primary human prostate cell strains (i.e., HPCA-10a and HPCA-10c) derived from high-grade cancer. Northern blots, Western blots, and ELISAs showed that IL-10 suppressed IGF-I induction of MMP-2 and MT1-MMP mRNA synthesis in these cell strains (P < 0.001). Inhibition studies with IL-10 and IGF-I receptor antibodies plus transfections experiments with IL-10 sense, and IGF-I receptor antisense constructs confirmed these results. Finally, transient transfection experiments and chloramphenicol acetyltransferase assays with different regions of the 5' promoter region of the MMP-2 gene (-1659 to -555 bp) additionally showed that IGF-I stimulated p53-dependent plasmid catecholamine acetyltransferase activity and that IL-10 blocked IGF-I-induced plasmid catecholamine acetyltransferase activity. Electrophoretic mobility shift assays revealed that IL-10 induced protein(s) binding to a putative "silencer element" ( 1309 to -555 fragment) downstream of the p53 binding site (-1649 to -1640). The data show that IL-10 blocks IGF-I activation of MMP-2 and MT1-MMP mRNA expression and protein synthesis in primary prostate cell strains. PMID- 12631627 TI - Antiangiogenesis treatment combined with chemotherapy produces chondrosarcoma necrosis. AB - A combination therapy protocol using a marine chemotherapeutic and an antiangiogenic molecule was tested in a mouse tumor xenograft model for the ability to curtail the growth of a human chondrosarcoma (CHSA). Ecteinascidin-743 (ET-743), a marine-derived chemotherapeutic, was effective at slowing the growth of a primary CHSA. Plasminogen-related protein B, which antagonizes various endothelial cell activities, also elicited a significant inhibition of neoplastic growth, albeit with reduced effectiveness. The combination of the two agents resulted in only a modest further repression of tumor growth over that associated with ET-743 treatment alone, as measured by tumor volume (82% versus 76% inhibition, respectively). However, analysis of the extent of tumor necrosis and vascularization of the tumor revealed that the coadministration of the two compounds was clearly more effective, eliciting a 2.5-fold increase in tumor necrosis relative to single-agent treatment. The combination therapy also was most effective at antagonizing tumor-associated microvessel formation, as assessed by CD31 immunostaining, suggesting that combination therapy may hold promise for treating CHSA. Tumor necrosis produced by combination therapy of ET 743 and recombinant plasminogen-related protein B was also significantly greater than that produced by conventional doxorubicin treatment, further corroborating the efficacy of combination therapy. PMID- 12631626 TI - Blockade of epidermal growth factor receptor signaling in tumor cells and tumor associated endothelial cells for therapy of androgen-independent human prostate cancer growing in the bone of nude mice. AB - PURPOSE: We determined whether blockade of the epidermal growth factor receptor (EGF-R) signaling pathway by oral administration of the EGF-R tyrosine kinase inhibitor (PKI 166) alone or in combination with injectable Taxol inhibits the growth of PC-3MM2 human prostate cancer cells in the bone of nude mice. EXPERIMENTAL DESIGN: Male nude mice implanted with PC-3MM2 cells in the tibia were treated with oral administrations of PKI 166 or PKI 166 plus injectable Taxol beginning 3 days after implantation. The incidence and size of bone tumors and destruction of bone were determined by digitalized radiography. Expression of epidermal growth factor (EGF), EGF-R, and activated EGF-R in tumor cells and tumor-associated endothelial cells was determined by immunohistochemistry. RESULTS: Oral administration of PKI 166 or PKI 166 plus injectable Taxol reduced the incidence and size of bone tumors and destruction of bone. Immunohistochemical analysis revealed that PC-3MM2 cells growing adjacent to the bone expressed high levels of EGF and activated EGF-R, whereas tumor cells in the adjacent musculature did not. Moreover, endothelial cells within the bone tumor lesions, but not in uninvolved bone or tumors in the muscle, expressed high levels of activated EGF-R. Treatment with PKI 166 and more so with PKI 166 plus Taxol significantly inhibited phosphorylation of EGF-R on tumor and endothelial cells and induced significant apoptosis and endothelial cells within tumor lesions. CONCLUSIONS: These data indicate that endothelial cells exposed to EGF produced by tumor cells express activated EGF-R and that targeting EGF-R can produce significant therapeutic effects against prostate cancer bone metastasis. PMID- 12631628 TI - Oncolytic viral therapy for human pancreatic cancer cells by reovirus. AB - PURPOSE: Pancreatic cancer has a poor prognosis and few effective therapies are available. The oncolytic effect of reovirus has been observed in cancer cells with an activated Ras signaling pathway, and pancreatic cancer may be a candidate target for reovirus because K-ras mutation is frequently found in pancreatic cancer. EXPERIMENTAL DESIGN: In this study, we examined the feasibility of using reovirus (serotype 3) as an antihuman pancreatic cancer agent. RESULTS: Reovirus was able to infect five human pancreatic cancer cell lines (Panc1, MIApaca-2, PK1, PK9, and BxPC3) in vitro. We also confirmed that the Ras activity in these cancer cell lines was elevated compared with that in the normal cell line and that susceptibility to reovirus was associated with the Ras activity of these cells. In a unilateral murine xenograft model using Panc1 and BxPC3 cell lines, each tumor growth was suppressed by intratumoral injection of reovirus. Furthermore, local injection of reovirus also had systemic antitumor effects in a bilateral xenograft model using Panc1 cell line. Immunohistochemical examination revealed that reovirus replication was observed within the tumor but not in surrounding normal tissue. CONCLUSIONS: These results suggest that reovirus can be considered for a novel therapy against pancreatic cancer. PMID- 12631629 TI - Correspondence re: T. Fehm et al., Cytogenetic evidence that circulating epithelial cells in patients with carcinoma are malignant. Clin. Cancer Res., 8: 2073-2084, 2002. PMID- 12631631 TI - Use of CT to assess the efficacy of an oncolytic adenovirus. PMID- 12631632 TI - Dr. Gary J. Becker Young Investigator Award: intraarterial adenovirus for metastatic gastrointestinal cancer: activity, radiographic response, and survival. AB - PURPOSE: To determine the antitumoral activity and radiographic response pattern of intraarterial administration of a selective replication-competent adenovirus in patients with hepatic metastases from gastrointestinal carcinomas. MATERIALS AND METHODS: Thirty-five patients were treated, seven in the dose-escalation phase and 28 at high doses. Inclusion criteria allowed mild laboratory value and performance status abnormalities and as much as 50% replacement of hepatic volume by tumor. An attenuated adenovirus that selectively replicates in p53-deficient cells (Onyx-015) was administered by hepatic arterial infusion at doses as high as 2 x 10(12) particles for two cycles. Subsequent cycles (maximum of eight total) were administered in combination with intravenous 5-fluorouracil (5-FU) and leucovorin. RESULTS: Tumor responses were demonstrated in combination with chemotherapy, even in 5-FU-resistant patients. The 15 patients who responded radiographically showed a pattern of acute tumor enlargement despite normalization of laboratory and clinical parameters, followed by very slow regression of tumor size. Radiographic response did not correlate with p53 status. Median survival of radiographic responders (475 days) was significantly longer than that of nonresponders (143 days). CONCLUSIONS: Hepatic arterial infusion of the replication-selective adenovirus Onyx-015 in combination with chemotherapy resulted in tumor regressions in select patients, including some in whom previous chemotherapy had failed. A biphasic radiographic response pattern was demonstrated. The mechanism of action appears to be more complex than that seen in vitro. PMID- 12631633 TI - Drug-eluting stents: potential applications for peripheral arterial occlusive disease. AB - Many different approaches have been evaluated to prevent restenosis in stents after vascular implantation. Currently, drug-eluting stents are extremely promising in suppressing neointimal hyperplasia. Various animal studies and randomized trials in humans have shown excellent results in terms of safety and efficacy during intermediate-term follow-up. This article will give an overview of experimental and clinical data of the different agents in published and ongoing trials. PMID- 12631635 TI - Angiography for acute lower gastrointestinal hemorrhage: efficacy of cut film compared with digital subtraction techniques. AB - PURPOSE: To find out if digital subtraction angiography (DSA) has a different diagnostic power than conventional cut-film angiography in investigating acute lower gastrointestinal (GI) hemorrhage. MATERIALS AND METHODS: Retrospective analysis of two cohorts of patients investigated by angiography for acute lower GI hemorrhage was performed. One group of patients treated at one center ("hospital H") underwent conventional angiography. The other patients, treated at another center ("hospital G"), were investigated exclusively by DSA. The groups were compared for demographic characteristics, etiology, and bleeding parameters. The diagnostic power of both angiographic modalities was compared and statistically analyzed. RESULTS: Fifty-five lower GI hemorrhages investigated by conventional cut-film angiography and 53 cases studied by DSA were included. Both study groups had statistically similar demographic and bleeding characteristics. Overall sensitivity, specificity, and positive and negative predictive values of cut-film angiography for localizing the lower GI bleeding were 62%, 73%, 86%, and 42%, whereas, for DSA, the respective values were 60%, 100%, 100%, and 24% (Z test P values were.78,.02,.03, and.16). CONCLUSIONS: DSA is as sensitive as conventional cut-film angiography for localizing acute lower GI bleeding. If a lesion is found, the chance that it will be the bleeding source is higher with DSA than with conventional angiography. PMID- 12631634 TI - Long-term results of ePTFE stent-graft versus angioplasty in the femoropopliteal artery: single center experience from a prospective, randomized trial. AB - PURPOSE: To describe a single-center experience as part of a U.S. multicenter prospective randomized trial of PTA versus percutaneous transluminal angioplasty (PTA)- and ePTFE (expanded polytetrafluoroethylene)-covered endoprosthesis placement for the treatment of superficial femoral artery (SFA) and proximal popliteal artery stenoses and occlusions. MATERIALS AND METHODS: Twenty-eight patients with claudication or ischemia were treated by PTA alone (n = 13) or PTA and endoprosthesis placement (n = 15). Baseline characteristics, including exercise ankle/brachial index (ABI), number of patent runoff vessels, Rutherford Becker ischemia score, and lesion length, were equivalent. Follow-up included postprocedure, objective, noninvasive vascular evaluation and a clinical status scale for a minimum of 24 months. RESULTS: Technical success was achieved in 15 of 15 patients (100%) in the endoprosthesis group and 12 of 13 patients (92%) in the PTA group. Complications in the endoprosthesis group included clinically significant embolization (n = 1, successfully treated by thrombolysis) and transient thigh pain that required medication in three patients (20%). Clinical improvement was achieved initially in all patients with a significant increase in exercise ABI in both groups (mean increase from baseline was 0.51 with 95% CI: 0.37-0.64 in endoprosthesis patients and 0.39 with 95% CI: 0.31-0.47 in PTA patients; P =.13). At 6-month follow-up with duplex US, 93% of patients (14 of 15) treated with the Hemobahn endoprosthesis (W.L. Gore & Associates, Flagstaff, AZ) remained primarily patent versus 42% of patients (five of 12) treated with PTA alone. At 2 years follow-up, primary patency remained 87% (13 of 15 patients) in the endoprosthesis group versus only 25% (three of 12 patients) in the PTA group (P =.002). Exercise ABI was >0.15 higher than baseline in 93% of patients (14 of 15) treated with endoprosthesis placement versus 54% of patients (seven of 13) treated by PTA alone. Clinical success was maintained in 87% of patients (13 of 15) in the endoprosthesis group versus 46% of patients (six of 13) in the PTA group. CONCLUSION: This single-center experience with placement of the Hemobahn endoprosthesis (W.L. Gore & Associates) in the SFA demonstrated a statistically significant improvement in both patency and clinical outcome compared with PTA alone. PMID- 12631636 TI - Transjugular liver biopsy in patients with hematologic malignancy and severe thrombocytopenia. AB - PURPOSE: The purposes of this study are to report experience with transjugular liver biopsy (TJLB) in patients with hematologic malignancy and severe thrombocytopenia and to determine the incidence of hemorrhage-related complications in patients with prebiopsy and pretransfusion platelet counts of 30 x 10(9) /L or lower to propose a threshold platelet count above which TJLB can be safely performed without transfusion. MATERIALS AND METHODS: Medical records and laboratory reports of 50 patients with severe thrombocytopenia who had undergone 51 TJLB procedures and prebiopsy platelet transfusions between August 1999 and September 2001 were retrospectively reviewed. Biopsy success and procedural complications were recorded. RESULTS: TJLB was technically successful in 49 of 51 procedures (96%). The mean prebiopsy, pretransfusion platelet count was 17 x 10(9)/L (range, 3-30 x 10(9)/L) and a mean of 11 U (range, 6-32 U) of platelets per patient were transfused. The overall mean postbiopsy platelet count was 38 x 10(9)/L (range, 5-105 x 10(9)/L), but it remained 30 x 10(9)/L or lower in 24 TJLB procedures. No hemorrhage-related complications were encountered, but ventricular fibrillation occurred in one patient during the procedure. CONCLUSION: A threshold platelet count for safe TJLB resides below 30 x 10(9)/L. A prospective study is necessary to better define a lower threshold above which TJLB can be performed without platelet transfusion. PMID- 12631637 TI - Renal perfusion defects after endovascular repair of abdominal aortic aneurysms. AB - PURPOSE: To determine the prevalence of new renal perfusion defects and the association of these events with aneurysm neck atheroma during endovascular stent graft repair of abdominal aortic aneurysm (AAA). MATERIALS AND METHODS: A retrospective review was performed of 50 patients who underwent AAA repair with bifurcated endovascular stent-grafts without suprarenal components between June 1998 and May 1999. Pre- and postprocedural computed tomographic (CT) angiograms were reviewed to determine the prevalence of new renal perfusion defects. The percent volume of atheroma of the aneurysm neck was determined by three dimensional volumetric reformation and correlated with prevalence of new postprocedural renal perfusion defects. Follow-up CT angiography was performed between 6 months and 2 years after the procedure and used to evaluate the presence of residual defects and interval changes. RESULTS: Of 50 subjects, 18% (n = 9) had new perfusion defects presumed to be embolic in origin. Follow-up was available for four patients: scarring and cortical thinning consistent with infarction developed in two, whereas the defects resolved in the other two. Analysis of aneurysm neck revealed an average percentage of atheroma of 32% and a range of 0%-73%. In subjects with >/=40% neck atheroma, the prevalence of new renal perfusion defects was 45.4% (five of 11), compared to 10.3% (four of 39) in subjects with <40% neck atheroma. This difference was significant (P =.0170). CONCLUSIONS: In this series, the frequency of renal embolic events associated with AAA endovascular repair was 18%. Prevalence of renal embolic perfusion defects was shown to correlate with volume of aneurysm neck atheroma. PMID- 12631638 TI - Pulse-inversion harmonic imaging improves lesion conspicuity during US-guided biopsy. AB - PURPOSE: To assess the feasibility of percutaneous biopsy of low-conspicuity focal liver lesions with use of pulse-inversion harmonic imaging (PIHI) guidance in the late phase after injection of microbubble contrast agent. MATERIALS AND METHODS: Twelve patients referred for ultrasound (US)-guided biopsy had liver lesions lacking adequate conspicuity to undergo biopsy under conventional ultrasound (US) guidance. They underwent biopsy procedures performed with the use of PIHI guidance in the late phase after injection of Levovist. The conspicuity of lesions, procedure time, number of passes, and success and complication rates were documented and compared to a control group. The control group consisted of retrospective analysis of 19 patients who had undergone conventional US-guided biopsy procedures performed by the same radiologist. RESULTS: After contrast material injection, all lesions had sufficient increases in conspicuity to be targeted under PIHI guidance. Procedure time was prolonged in the PIHI group (66 minutes; range, 30-120 min; vs control, 33 min; range, 15-77 min; P <.01, Wilcoxon signed-rank test). The number of passes was greater in the PIHI group (1.9 +/- 0.3; vs control, 1.4 +/- 0.6; P <.01, chi(2) test). No statistically significant difference was observed when comparing success and complication rates. CONCLUSION: PIHI in the late phase after injection of Levovist generated clear delineation of focal liver lesions in all cases, allowing an accurate and safe sampling. This technique broadens the scope of US-guided biopsy to lesions lacking adequate conspicuity on conventional US. PMID- 12631639 TI - Microbubble-augmented ultrasound declotting of thrombosed arteriovenous dialysis grafts in dogs. AB - PURPOSE: Transcutaneous low-frequency ultrasound (LFUS) can effectively lyse clots in the presence of microbubbles. This study was designed to test the commercially available human albumin microspheres injectable suspension octafluoropropane formulation, Optison, to establish efficacy and assess US parameters of intensity and wave modes in a canine model of a thrombosed arteriovenous (dialysis) graft. MATERIALS AND METHODS: Arteriovenous grafts in five dogs were cannulated, temporarily ligated, and thrombosed. Different declotting techniques were randomized to treat nine groups. Control groups involved direct saline (4.5 mL) clot injection in 0.5-1.0-mL increments. One group underwent peripheral intravenous microbubble injection (13.5 mL). Six groups underwent direct incremental clot injection of 4.5 mL of microspheres with LFUS for 30 minutes in 3-5-minute increments with use of various intensity settings in continuous-wave and pulsed-wave (PW) modes. At each increment, angiography was used to grade flow, declotting, and overall success. RESULTS: One hundred four procedures showed success in all 24 high-intensity PW modes (1.2-2.0 W/cm(2)); only one of 20 control experiments was successful (P <.0001). Medium intensity modes yielded intermediate success rates. Lowest-intensity direct injection groups and intravenous and control groups ranked lower. Results at 30 minutes were better than at 15 minutes (P <.0001). CONCLUSIONS: LFUS with direct injection of microbubbles is effective in lysing moderate-sized clots and recanalizing thrombosed arteriovenous grafts. It best succeeds at the higher range of intensity settings tested in PW mode. Further development is justified. PMID- 12631640 TI - Remodeling of suspended small intestinal submucosa venous valve: an experimental study in sheep to assess the host cells' origin. AB - PURPOSE: To investigate the origin of host cells during remodeling of small intestinal submucosa (SIS) square stent-based bicuspid venous valves (VVs). MATERIALS AND METHODS: Suspended VVs (SVVs) were developed by suspending VVs within bare square stents so the valve elements would not contact the vein wall after deployment. Eight SVVs were placed within the intrahepatic and infrahepatic inferior venae cavae (IVCs) of four adult female sheep. Eight standard VVs were implanted in the external jugular veins of these animals for comparison. At 5 weeks after placement, the devices were examined for stability and patency and the animals were killed. Gross, histologic, and scanning electron microscopic (SEM) examinations were performed. RESULTS: Follow-up spot radiographs and venography showed no migration of the devices, venous occlusion, or thrombus formation. All SVVs were intact without contact with the IVC wall. Six VVs were competent and two were slightly tilted with some reflux. Histologic study showed remodeling of SVVs and VVs with newly formed collagen fibers; fibroblasts and inflammatory cells were found penetrating the SIS leaflets and endothelial cells on the surface. SIS neovascularization was also present. There was no difference regarding SIS remodeling between SVVs and the free part of VV leaflets. The VV leaflets' bases were thicker compared to their free parts (P <.01). SEM examination showed endothelial cells on both sides of the SVVs and VVs. Endothelialization of the SVV central leaflet surfaces and both surfaces of the VV leaflets was more complete than that of the peripheral surfaces of the SVV leaflets. CONCLUSION: SIS-based valve remodeling occurs independently of vessel wall contact by recruitment of cells directly from the circulation. PMID- 12631641 TI - Metallic stent placement in the pancreatic duct: an experimental study in the normal dog pancreas. AB - PURPOSE: To evaluate radiologic and histologic changes in the pancreatic duct and parenchyma after placement of metallic stents in normal dog pancreatic ducts. MATERIALS AND METHODS: Eight dogs underwent duodenotomy and placement of a balloon-expandable metallic stent 1.5 mm in diameter and 1 cm in length in the main pancreatic duct. Two dogs each were killed at 1 day and 1, 3, and 5 months after stent placement. Pancreatic duct injection images were obtained before and immediately after stent placement and just before and after sacrifice. Macroscopic and microscopic examinations were also performed. RESULTS: Pancreatic duct injection images revealed mild dilation of pancreatic ducts related to the procedure immediately and 1 day after stent placement. Microscopically, denudation of the ductal epithelium and small foci of fat necrosis were observed. At 1 month or later, ductal stricture (n = 4) and occlusion (n = 2) were found at the downstream edge of the initially stent-implanted region associated with dilation of the duct upstream. Epithelial hyperplasia was observed at the sites of ductal stricture and occlusion at 1 and 3 months. Stents remained in the region in which they were initially placed until 1 month. However, stents had moved downstream from their initial positions at 3 and 5 months. One stent had migrated into the duodenum at 5 months. Stone formation was found in the ductal lumen and around the stents at 3 and 5 months. Ducts and parenchyma became fibrotic proportional to the period of stent implantation. CONCLUSION: Metallic stent placement in the pancreatic duct causes ductal and parenchymal damage related to the procedure and reaction to the stent in the normal dog pancreas. PMID- 12631643 TI - Management of TIPS-related refractory hepatic encephalopathy with reduced Wallgraft endoprostheses. AB - The purpose of this study is to evaluate the feasibility of constrained endografts used for the treatment of transjugular intrahepatic portosystemic shunt (TIPS)-related refractory hepatic encephalopathy (HE). Because the clinical status of two patients worsened (return of intractable ascites requiring transplantation, n = 1; death, n = 1) after complete balloon occlusion, six patients were treated with constrained/modified Wallgraft endoprostheses placed within the preexisting TIPS. Shunt reductions were technically successful in all six patients, as shown by an immediate mean portosystemic gradient increase of 9.3 mm Hg. Clinical improvement was achieved in five patients within 72 hours of reduction. The remaining patient continued to decline and died 3 weeks later. Two endografts completely occluded within 8 months without HE recurrence. This technique offers an attractive alternative to previously described shunt reduction methods. PMID- 12631642 TI - Transcatheter arterial embolization of pulmonary sequestration in neonates: long term follow-up results. AB - The purposes of this study are to describe experience with the safety and long term efficacy of transcatheter arterial embolization (TAE) of pulmonary sequestration in neonates and to introduce a new technique of embolization by an umbilical-artery route. TAE was performed in five neonates, via the femoral artery in one and the umbilical artery in four. Complete regression was achieved in four cases and partial regression (>90%) was obtained in one. TAE is a safe and effective alternative therapeutic modality for the treatment of pulmonary sequestration. The umbilical artery represents a preferable route for performing embolization in neonates. PMID- 12631644 TI - Image-guided percutaneous radiofrequency ablation for osteoid osteomas. AB - The authors present a retrospective analysis of the technical and clinical successes, complications, and clinical follow-up of image-guided percutaneous radiofrequency (RF) ablation of osteoid osteomas. Nine patients with osteoid osteomas underwent image-guided localization of osteoid osteomas. Outpatient percutaneous therapy (13 procedures) was performed under general anesthesia after image-guided localization of the nidus. Initial technical success was achieved in seven of nine patients. Two initial technical and clinical failures occurred early in this experience because of failure to adequately enter the nidus with use of fluoroscopic imaging alone. Clinical success was achieved in eight of nine patients. No major immediate or delayed complications were observed. PMID- 12631645 TI - Filter-related, thrombotic occlusion of the inferior vena cava treated with a Gianturco stent. AB - The authors report a case of complete thrombotic occlusion of the inferior vena cava (IVC), which occurred 4 weeks after placement of an IVC filter (TrapEase; Cordis, Miami FL). Initial treatment with suction thrombectomy and thrombolysis was ineffective. Percutaneous removal of the filter was unsuccessful because of the long period of implantation. TrapEase filters (Cordis) are easily collapsible because of their symmetric design and composition (nitinol). An expandable metallic Gianturco Z stent (Cook, Bloomington, IN) was used to exclude the filter from the vessel lumen. In cases of persistent filter-related, thrombotic occlusion of the IVC, in which initial treatment has failed, the use of a Gianturco stent (Cook) to exclude the filter from the vessel lumen is a viable treatment option if the filter has a collapsible design. PMID- 12631647 TI - Biliary ascariasis: percutaneous transhepatic management. AB - In a 60-year-old woman who underwent cholecystectomy and T-tube drainage for biliary ascariasis, postoperative bilirubin levels continued to increase. T-tube cholangiography revealed multiple filling defects in the bile ducts, which suggested roundworm reinfestation. Because the patient could not tolerate endoscopy, a percutaneous transhepatic approach with balloon dilatation of the ampulla of Vater and subsequent advancement of roundworms into the duodenum with an embolectomy balloon was used. The procedure was successful, with no major complications. The use of a percutaneous transhepatic treatment to advancement of roundworms into the duodenum is a feasible alternative to surgery when endoscopic extraction fails. PMID- 12631646 TI - Retroperitoneal hematoma caused by a ruptured pelvic varix in a patient with iliac vein compression syndrome. AB - May-Thurner syndrome, or iliac vein compression syndrome (IVCS), is a rare but well-described entity. It refers to the formation of venous thrombus caused by compression of the left iliac vein, most commonly between the right iliac artery and lumbar vertebrae. Several variants of IVCS have been described, including unusual presenting symptoms, etiologies, and complications. The authors describe an unusual case of IVCS in which the patient presented with a left-sided retroperitoneal hematoma arising from a ruptured collateral venous varix shortly after the development of symptomatic left lower-extremity deep vein thrombosis. PMID- 12631648 TI - Precipitation of gadolinium and ethanol during nerve block. PMID- 12631649 TI - Is fracture risk increased in patients with coeliac disease? PMID- 12631650 TI - Crohn's disease or Crohn's diseases? PMID- 12631651 TI - Transplantation for alcoholic liver disease: lessons from the explant? PMID- 12631652 TI - Effect of acute and chronic administration of the GABA B agonist baclofen on 24 hour pH metry and symptoms in control subjects and in patients with gastro oesophageal reflux disease. AB - BACKGROUND AND AIMS: The gamma-aminobutyric acid (GABA(B)) agonist baclofen has been shown to reduce reflux episodes during the first three postprandial hours in patients with gastro-oesophageal reflux disease (GORD) and in normal controls. The aim of the study was to assess the effect of acute (one day) and chronic (four weeks) administration of baclofen on 24 hour pH metry and symptoms in GORD patients and normal controls. PATIENTS AND METHODS: Acute study: 28 patients with GORD with none or mild oesophagitis at endoscopy and 15 controls underwent oesophageal and gastric 48 hour pH metry in which baclofen or placebo was given for 24 hours in a double blinded manner. Chronic study: 16 GORD patients received baclofen (10 mg four times daily) or placebo for four weeks. Twenty four hour oesophageal pH metry and reflux symptom scores were evaluated before and at the end of treatment. RESULTS: Acute study: the number of reflux episodes and per cent time with pH <4 was significantly lower after baclofen in GORD patients and controls (p<0.003; p<0.0007). Gastric pH increased significantly in GORD patients and controls (p<0.001; p<0.05). Chronic study: four weeks after initial administration of baclofen, the number of reflux episodes and percentage of time with pH <4 significantly decreased in all GORD patients (p<0.003; p<0.02). Symptom scores significantly improved after treatment with baclofen (p<0.0007). CONCLUSIONS: The GABA(B) agonist baclofen reduces 24 hour gastro-oesophageal reflux and increases gastric pH in GORD patients and controls. When given for one month to GORD patients, baclofen reduces oesophageal acid refluxes and significantly improves symptoms. Baclofen may be useful in the therapy of GORD. PMID- 12631653 TI - Biomechanics, diagnosis, and treatment outcome in inflammatory myopathy presenting as oropharyngeal dysphagia. AB - AIMS: In patients with inflammatory myopathy and dysphagia, our aims were to determine: (1) the diagnostic utility of clinical and laboratory indicators; (2) the biomechanical properties of the pharyngo-oesophageal segment; (3) the usefulness of pharyngeal videomanometry in distinguishing neuropathic from myopathic dysphagia; and (4) clinical outcome. METHODS: Clinical, laboratory, and videomanometric assessment was performed in 13 patients with myositis and dysphagia, in 17 disease controls with dysphagia (due to proven CNS disease), and in 22 healthy age matched controls. The diagnostic accuracy of creatine kinase (CPK), erythrocyte sedimentation rate, antinuclear antibody, and electromyography (EMG) were compared with the gold standard muscle biopsy. The biomechanical properties of the pharyngo-oesophageal segment were assessed by videomanometry. RESULTS: Mean time from dysphagia onset to the diagnosis of myositis was 55 months (range 1-180). One third had no extrapharyngeal muscle weakness; 25% had normal CPK, and EMG was unhelpful in 28%. Compared with neurogenic controls, myositis patients had more prevalent cricopharyngeal restrictive disorders (69% v 14%; p=0.0003), reduced upper oesophageal sphincter (UOS) opening (p=0.01), and elevated hypopharyngeal intrabolus pressures (p=0.001). Videomanometric features favouring a myopathic over a neuropathic aetiology were: preserved pharyngeal swallow response, complete UOS relaxation, and normal swallow coordination. The 12 month mortality was 31%. CONCLUSIONS: The notable lack of supportive clinical signs and significant false negative rates for laboratory tests contribute to the marked delay in diagnosis. The myopathic process is strongly associated with restricted sphincter opening suggesting that cricopharyngeal disruption is a useful adjunct to immunosuppressive therapy. The condition has a poor prognosis. PMID- 12631654 TI - Delivery of radiofrequency energy to the lower oesophageal sphincter and gastric cardia inhibits transient lower oesophageal sphincter relaxations and gastro oesophageal reflux in patients with reflux disease. AB - BACKGROUND AND AIMS: Radiofrequency energy (RFe) treatment to the lower oesophageal sphincter (LOS) and gastric cardia is a new luminally delivered therapy proposed as an alternative treatment for gastro-oesophageal reflux disease (GORD). However, it is unclear how RFe achieves its antireflux effect. This study investigated the effects of RFe on mechanisms of spontaneous reflux in patients with GORD. METHODS: Twenty patients with GORD underwent endoscopy, symptom evaluation, and combined postprandial oesophageal manometry and pH monitoring before and six months after RFe, and 24 hour ambulatory pH monitoring before and at six and 12 months after treatment. RESULTS: RFe reduced the rate of postprandial transient LOS relaxations from 6.8 (5.7-8.1) (median (interquartile range) per hour to 5.2 (4.2-5.8) per hour (p<0.01), and increased mean basal LOS pressure from 5.2 (SEM 0.3) mm Hg to 8.0 (SEM 0.4) mm Hg (p<0.01). The number of reflux events was reduced from 10 (2-15.3)/3 hours to 5 (3.5-8.5)/3 hours (p<0.05) and there was an associated significant reduction in acid exposure time from 5.4% (0.4-14.7) to 3.9% (0.4-6.6) (p<0.05). RFe significantly reduced ambulatory oesophageal acid exposure from 10.6% (7.8-13.0) to 6.8% (3.1-9.1) (p<0.01) at six months and 6.3% (4.7-10.9) (p<0.05) at 12 months. All patients required acid suppressant medication for symptom control before RFe. Six months after treatment, 15 patients (75%) were in symptomatic remission and 13 (65%) at 12 months. CONCLUSIONS: RFe has significant effects on LOS function that are associated with improvement in the antireflux barrier. Uncontrolled clinical data also suggest a beneficial effect in the control of reflux symptoms in these patients. PMID- 12631655 TI - Can extent of high grade dysplasia in Barrett's oesophagus predict the presence of adenocarcinoma at oesophagectomy? AB - BACKGROUND: Optimal management of Barrett's oesophagus complicated by high grade dysplasia is controversial. Recently, the extent of high grade dysplasia was described as a predictor of subsequent development of cancer in patients undergoing continued surveillance. However, there is no universal agreement on the definition of extent of high grade dysplasia. AIM: To determine if extent of high grade dysplasia in Barrett's oesophagus is a predictor of the presence of adenocarcinoma at the time of oesophagectomy. METHODS: Forty two patients with Barrett's oesophagus and high grade dysplasia who underwent oesophagectomy between 1985 and 1999 were identified from a prospective database. All pathological specimens, including preoperative endoscopic biopsies and post oesophagectomy sections, were reviewed in a blinded fashion by one expert gastrointestinal pathologist to determine the extent of high grade dysplasia. The extent of high grade dysplasia was defined using two different criteria, one from the Cleveland Clinic and one from the Mayo Clinic. RESULTS: Twenty four of 42 patients (57%) had unsuspected cancer at the time of oesophagectomy. Using the Cleveland Clinic definition, 10 of 21 (48%) patients with focal high grade dysplasia had carcinoma compared with 14 of 21 patients (67%) with diffuse high grade dysplasia (p=0.35). Using the Mayo Clinic definition, adenocarcinoma was found in five of seven (72%) patients with focal high grade dysplasia compared with 19 of 35 (54%) with diffuse high grade dysplasia (p=0.68). CONCLUSIONS: The extent of high grade dysplasia, regardless of how it is defined, does not predict the presence of unsuspected adenocarcinoma at oesophagectomy. There is no evidence as yet that the extent of high grade dysplasia can be used as a basis for decision making in these patients. PMID- 12631656 TI - Aspirin inhibits the growth of Helicobacter pylori and enhances its susceptibility to antimicrobial agents. AB - BACKGROUND AND AIM: The role of Helicobacter pylori and aspirin in peptic ulcer formation and recurrence remains an important clinical topic. The interaction between aspirin and H pylori in vitro is also not clear. We investigated the effect of aspirin on the growth of H pylori and on the susceptibility of H pylori to antimicrobials. METHODS: Time killing studies of H pylori were performed with different concentrations of aspirin and salicylate. Growth of bacteria was assessed spectrophotometrically and by viable colony count. The effects of aspirin on the efficiency of colony formation and on metronidazole induced mutation to rifampicin resistance in H pylori were determined. Minimal inhibitory concentrations (MICs) of aspirin and metronidazole were tested by the standard agar dilution method. MICs of amoxycillin and clarithromycin were determined by the E test method. RESULTS: Aspirin and salicylate inhibited the growth of H pylori in a dose dependent manner and bactericidal activity was due to cell lysis. Aspirin 400 micro g/ml caused a 2 logs decrease in colony forming units/ml at 48 hours, and suppressed the normal ability of metronidazole to induce new mutations to rifampicin. The IC(90) of aspirin was 512 micro g/ml. Increased susceptibility of amoxycillin, clarithromycin, and metronidazole to H pylori was observed at 1 mM (180 micro g/ml) aspirin. CONCLUSIONS: Aspirin inhibited the growth of H pylori, suppressed the mutagenic effect of metronidazole, and enhanced the susceptibility of H pylori to antimicrobial agents. This mechanism is important in future drug development for effective clearing and overcoming resistance. PMID- 12631657 TI - Concomitant alterations in intragastric pH and ascorbic acid concentration in patients with Helicobacter pylori gastritis and associated iron deficiency anaemia. AB - BACKGROUND: Seroepidemiological and clinical studies suggest that Helicobacter pylori may cause iron deficiency anaemia (IDA) in the absence of peptic lesions by undefined mechanisms, which still remain to be fully elucidated. Gastric acidity and ascorbic acid (AA) promote iron absorption. AA is lowered in the presence of H pylori infection. H pylori can cause atrophic body gastritis with achlorhydria, decreased iron absorption, and consequent IDA. Whether alterations in intragastric acidity and AA concentrations play a role in IDA developing in patients with H pylori gastritis remains to be determined. AIM: To evaluate gastric juice pH and gastric juice and plasma AA in patients with H pylori infection and unexplained IDA, compared with controls with IDA and a healthy stomach or with controls with H pylori infection and no IDA. RESULTS: Patients with IDA and H pylori gastritis were characterised by concomitant increased intragastric pH (median value 7) and decreased intragastric AA (median value 4.4 micro g/ml) compared with controls with a healthy stomach (median pH 2; median intragastric AA 17.5 micro g/ml) and with H pylori positive controls without IDA (median pH 2.1; median intragastric AA 7.06 micro g/ml). Intragastric AA was inversely related to pH (r=-0.40, p=0.0059) and corporal degree of gastritis (r= 0.53, p=0.0039). Plasma AA concentrations were lower in all infected groups than in healthy controls. CONCLUSIONS: Patients with unexplained IDA and H pylori gastritis present concomitant changes in intragastric pH and AA that may justify impaired alimentary iron absorption and consequent IDA. PMID- 12631658 TI - Promoter methylation of E-cadherin gene in gastric mucosa associated with Helicobacter pylori infection and in gastric cancer. AB - BACKGROUND: E-cadherin is an adhesion molecule involved in tumour invasion/metastasis. Silencing of E-cadherin by promoter CpG methylation has been shown in both familial and sporadic gastric cancers. Helicobacter pylori is a class I carcinogen in gastric cancer. AIMS: This study was undertaken to investigate the association between methylation of E-cadherin and H pylori in gastric mucosa from dyspeptic patients, and in intestinal metaplasia and primary and metastatic adenocarcinoma from surgical specimens of patients with gastric cancer. METHODS: E-cadherin methylation was studied using methylation specific polymerase chain reaction in microdissected tissue from biopsies or surgical resection specimens. E-cadherin expression was studied by immunohistochemistry. RESULTS: E-cadherin methylation was present in 31% (11/35) of gastric mucosae from dyspeptic patients, and was associated with H pylori infection (p=0.002), but was independent of the age of the patient or presence or absence of gastritis. E-cadherin methylation was present in 0% (0/8) of normal mucosa, 57% (12/21) of intestinal metaplasias, and 58% (15/26) of primary and 65% (21/32) of metastatic cancers. E-cadherin methylation status was concordant in 92% (11/12) of intestinal metaplasias and primary cancers, and in 85% (17/20) of primary and metastatic cancers from the same resected specimen. E-cadherin methylation in gastric cancer was associated with depth of tumour invasion (p=0.02) and regional nodal metastasis (p=0.05). CONCLUSION: E-cadherin methylation is an early event in gastric carcinogenesis, and is initiated by H pylori infection. PMID- 12631659 TI - Five years of complete remission of gastric diffuse large B cell lymphoma after eradication of Helicobacter pylori infection. AB - Long term follow up data are not available for cases of diffuse large B cell gastric lymphoma treated by eradicating Helicobacter pylori alone. We present the case of an 82 year old man with diffuse large B cell lymphoma localised to the stomach which responded to H pylori eradication and which has not recurred after more than five years of close follow up. Our patient was not a candidate for other modalities of treatment. This case demonstrates that the option of treating H pylori infection as the initial trial of treatment for localised diffuse large B cell lymphoma is appropriate for consideration. If medical therapy using eradication of H pylori is used, it is essential that the patient undergoes close observation and repeated surveillance endoscopies. PMID- 12631660 TI - Duodenal mucosal reductase in wild-type and Hfe knockout mice on iron adequate, iron deficient, and iron rich feeding. AB - BACKGROUND: Genetic haemochromatosis is a common hereditary iron loading disorder in humans. The disease is associated with loss of function mutations in the HFE gene. This is thought to change iron stores via increased iron absorption. AIMS: In this study we investigated how adaptation of mucosal reductase activity is engaged in this process and how the changes compare with adaptation seen when an iron deficient diet is fed. METHODS: Duodenal mucosal surface reductase was measured with nitroblue tetrazolium in age matched groups of male Hfe knockout mice (Hfe) and wild- type mice fed a purified diet containing normal (iron adequate), high (iron rich), or low (iron deficient) iron concentrations. RESULTS: Reductase activity increased when mice were fed an iron deficient diet and decreased when they were fed an iron rich diet. Total villus activity, as measured by the average area under the activity curve along the crypt-villus axis, was increased 2.8-2.9-fold by iron deficiency in both genotypes. Approximately half of this difference was attributable to the significantly increased length of the villi in mice on an iron deficient diet (p<0.05). Hfe knockout did not affect villus length but increased mucosal reductase activity near the villus tips. Similar increases (1.3-1.6-fold) were seen on all diets but the increase was significant for iron deficient and iron loaded diets only (p<0.05). CONCLUSION: Hfe gene product and dietary iron downregulate villus reductase activity in mice. PMID- 12631661 TI - Reflux oesophagitis in adult coeliac disease: beneficial effect of a gluten free diet. AB - BACKGROUND: Coeliac disease patients show a number of gastrointestinal motor abnormalities, including a decrease in lower oesophageal sphincter pressure. The prevalence of endoscopic oesophagitis in these subjects however is unknown. AIM: To evaluate whether untreated adult coeliac patients had an increased prevalence of reflux oesophagitis and, if so, to assess whether a gluten free diet exerted any beneficial effect on gastro-oesophageal reflux disease (GORD) symptoms. PATIENTS AND METHODS: We retrospectively studied 205 coeliac patients (females/males 153/52, median age 32 years) who underwent endoscopy for duodenal biopsy and 400 non-coeliac subjects (females/males 244/156, median age 37 years) referred for endoscopy for upper gastrointestinal symptoms. Each patient was given a questionnaire for evaluation of GORD symptoms prior to and 4-12 months after endoscopy. Coeliac patients were given a gluten free diet. Oesophagitis patients of both groups, following an eight week course of omeprazole, were re evaluated for GORD symptoms at four month intervals up to one year. Significance of differences was assessed by Fisher's exact test. RESULTS: Oesophagitis was present in 39/205 (19%, 95% confidence interval (CI) 13.8-25.0%) coeliac patients and in 32/400 (8%, 95% CI 5.5-11.1%) dyspeptic subjects. At the one year follow up, GORD symptoms relapsed in 10/39 (25.6%, 95% CI 13-42.1%) coeliacs with oesophagitis and in 23/32 (71.8%, 95% CI 53.2-86.2%) non-coeliac subjects with oesophagitis. CONCLUSION: Coeliac patients have a high prevalence of reflux oesophagitis. That a gluten free diet significantly decreased the relapse rate of GORD symptoms suggests that coeliac disease may represent a risk factor for development of reflux oesophagitis. PMID- 12631662 TI - Fracture experience of patients with coeliac disease: a population based survey. AB - BACKGROUND: While coeliac disease is now recognised as being associated with both osteoporosis and osteomalacia, the size of any increase in the risk of fracture in patients with coeliac disease compared with the general population has not been quantified. AIM: To examine the fracture experience of adults with coeliac disease compared with the general population. SUBJECTS: Patients with coeliac disease diagnosed in adulthood and born before 1950, selected from two large population based disease registers, and age and sex frequency matched controls identified from local general practitioner lists. METHODS: A four page lifestyle and general health questionnaire which included specific questions about fracture experience. RESULTS: Analysis was performed on 244 patients with coeliac disease and 161 controls, giving response rates of 89% and 72%, respectively. Eighty two (35%) coeliac patients and 53 (33%) controls reported ever having sustained one or more fractures, giving an age and sex adjusted odds ratio of 1.05 (95% confidence interval (CI) 0.68-1.62). The most common fracture site reported was the forearm or wrist, with an adjusted odds ratio of 1.21 (95% CI 0.66-2.25) for patients with coeliac disease having had a forearm or wrist fracture. Low trauma fractures were reported by 37 patients with coeliac disease (15.7%) and by 21 controls (13.8%), with an adjusted odds ratio of 1.16 (95% CI 0.65-2.10). The risk of low trauma fracture was slightly higher in coeliac men than women (odds ratio 1.28 compared with 1.12), but this difference was not statistically significant (p=0.84). After adjustment for age, sex, body mass index, and smoking status, patients with coeliac disease reported 13% more low trauma fractures than controls (odds ratio 1.13, 95% CI 0.60-2.12). There was no difference in low trauma fracture risk before and after diagnosis of coeliac disease. CONCLUSION: No overall increased fracture risk in patients with coeliac disease was observed. Although severe osteoporosis may develop in a subset of patients, as a whole patients with coeliac disease do not represent a population at particularly high risk of osteoporotic fracture and thus targeting them for osteoporosis screening and treatment is not justified. PMID- 12631663 TI - Increased rectal mucosal expression of interleukin 1beta in recently acquired post-infectious irritable bowel syndrome. AB - BACKGROUND AND AIMS: Chronic bowel disturbances resembling irritable bowel syndrome (IBS) develop in approximately 25% of patients after an episode of infectious diarrhoea. Although we have previously shown that psychosocial factors operating at the time of, or prior to, the acute illness appear to predict the development of post-infectious IBS (PI-IBS), our finding of an increased inflammatory cell number in the rectum persisting for at least three months after the acute infection suggested that there is also an organic component involved in the development of PI-IBS. To evaluate this further, we measured expressions of interleukin 1beta (IL-1beta) and its receptor antagonist (IL-1ra) in these patients to provide additional evidence that the pathogenesis of PI-IBS is underpinned by an inflammatory process. METHODS: Sequential rectal biopsy samples were prospectively obtained during and three months after acute gastroenteritis, from eight patients who developed post-infectious IBS (INF-IBS) and seven patients who returned to normal bowel habits after acute gastroenteritis (infection controls, INF-CON). Eighteen healthy volunteers who had not suffered from gastroenteritis in the preceding two years served as normal controls (NOR CON). IL-1beta and IL-1ra gene expressions were assayed by reverse transcriptase polymerase chain reaction, and their levels of expression were quantitated by optical densitometry after electrophoresis on agarose gel. RESULTS: INF-IBS patients exhibited significantly greater expression of IL-1beta mRNA in rectal biopsies than INF-CON patients both during and three months after acute gastroenteritis. Moreover, IL-1beta mRNA expression had increased in biopsies taken from INF-IBS patients at three months after the acute infection but no consistent change was observed in INF-CON patients. IL-1beta mRNA expression of INF-IBS patients at three months post gastroenteritis was significantly greater than NOR-CON whereas that of INF-CON patients was not significantly different from NOR-CON. Despite these differential changes in IL-1beta mRNA expression, no significant changes were observed in IL-1ra mRNA expression among the three groups. CONCLUSIONS: These findings indicate that those patients who develop IBS post infection exhibit greater IL-1beta mRNA expression, both during and after the infection, compared with individuals who do not develop PI-IBS. We conclude that such patients may be susceptible to inflammatory stimuli, and that inflammation may play a role in the pathogenesis of PI-IBS. PMID- 12631664 TI - COX-2 dependent inflammation increases spinal Fos expression during rodent postoperative ileus. AB - BACKGROUND AND AIMS: Cyclooxygenase 2 (COX-2) and prostaglandins (PGs) participate in the pathogenesis of inflammatory postoperative ileus. We sought to determine whether the emerging neuronal modulator COX-2 plays a significant role in primary afferent activation during postoperative ileus using spinal Fos expression as a marker. METHODS: Rats, and COX-2(+/+) and COX-2(-/-) mice underwent simple intestinal manipulation. The effect of intestinal manipulation on Fos immunoreactivity (IR) in the L(5)-S(1) spinal cord, in situ circumference, and postoperative leucocytic infiltrate of the intestinal muscularis was measured. Postoperative PGE(2) production was measured in peritoneal lavage fluid. The dependence of these parameters on COX-2 was studied in pharmacological (DFU, Merck- Frosst, selective COX-2 inhibitor) and genetic (COX-2(-/-) mice) models. RESULTS: Postoperative Fos IR increased 3.7-fold in rats and 2.2-fold in mice. Both muscularis leucocytic infiltrate and the circumference of the muscularis increased significantly in rats and COX-2(+/+) mice postoperatively, indicating dilating ileus. Surgical manipulation markedly increased PGE(2) levels in the peritoneal cavity. DFU pretreatment and the genetic absence of COX-2(-/-) prevented dilating ileus, and leucocytic infiltrate was diminished by 40% with DFU and by 54% in COX-2(-/-) mice. DFU reversed postsurgical intra- abdominal PGE(2) levels to normal. Fos IR after intestinal manipulation was attenuated by approximately 50% in DFU treated rats and in COX-2(-/-) mice. CONCLUSIONS: Postoperatively, small bowel manipulation causes a significant and prolonged increase in spinal Fos expression, suggesting prolonged primary afferent activation. COX-2 plays a key role in this response. This activation of primary afferents may subsequently initiate inhibitory motor reflexes to the gut, contributing to postoperative ileus. PMID- 12631665 TI - Isolation of peptides useful for differential diagnosis of Crohn's disease and ulcerative colitis. AB - BACKGROUND: Phage displayed random peptide technology has been utilised to identify binding epitopes of antibodies or receptor ligands. AIM: To isolates peptides from a phage library which are specifically recognised by antibodies in serum from patients with Crohn's disease (CD). METHODS: A phage displayed random peptide library composed of nine amino acids was established and sequentially screened using serum immunogloblin G obtained from CD patients. RESULTS: Five different CD specific peptides were isolated from the phage library. No homology in amino acid sequences was observed among four (CDP-1, -3 to -5) of the five peptides exhibiting different binding characteristics with each CD patient's serum. In contrast, two peptides (CDP-1 and -2) had similar amino acid sequences and similar binding characteristics. Four multiple antigenic peptides (MAP, CDP 1, -3 to -5) were synthesised, and an enzyme linked immunosorbent assay (ELISA) using the four peptides was developed to detect serum antibodies against them. Fifty two of 92 CD patients (56.5%) were detected by ELISA, none of 20 ulcerative colitis (UC) patients, only one of 25 duodenal ulcer patients, and only three of 48 healthy subjects. CONCLUSIONS: ELISA using the four peptides isolated in this study may be useful for the differential diagnosis of CD and UC. PMID- 12631666 TI - Analysis of the IBD5 locus and potential gene-gene interactions in Crohn's disease. AB - BACKGROUND AND AIMS: Genetic variation in the chromosome 5q31 cytokine cluster (IBD5 risk haplotype) has been associated with Crohn's disease (CD) in a Canadian population. We studied the IBD5 risk haplotype in both British and Japanese cohorts. Disease associations have also been reported for CARD15/NOD2 and TNF variants. Complex interactions between susceptibility loci have been shown in animal models, and we tested for potential gene-gene interactions between the three CD associated loci. METHODS: Family based association analyses were performed in 457 British families (252 ulcerative colitis, 282 CD trios) genotyped for the IBD5 haplotype, common CARD15, and TNF-857 variants. To test for possible epistatic interactions between variants, transmission disequilibrium test analyses were further stratified by genotype at other loci, and novel log linear analyses were performed using the haplotype relative risk model. Case control association analyses were performed in 178 Japanese CD patients and 156 healthy controls genotyped for the IBD5 haplotype. RESULTS: The IBD5 haplotype was associated with CD (p=0.007), but not with UC, in the British Caucasian population. The CARD15 variants and IBD5 haplotype showed additive main effects, and in particular no evidence for epistatic interactions was found. Variants from the IBD5 haplotype were extremely rare in the Japanese. CONCLUSIONS: The IBD5 risk haplotype is associated with British CD. Genetic variants predisposing to CD show heterogeneity and population specific differences. PMID- 12631667 TI - Genetic polymorphisms in biotransformation enzymes in Crohn's disease: association with microsomal epoxide hydrolase. AB - BACKGROUND: Mucosal biotransformation enzymes can modify toxic compounds in the gut. As chemical or oxidative stress may be involved in the aetiology of Crohn's disease, genes encoding for enzymes involved in the prevention of such stress may be candidates for genetic susceptibility to Crohn's disease. AIM: To assess the association of Crohn's disease with genetic polymorphisms in cytochrome P450 1A1, glutathione S-transferases mu-1, pi-1, and theta-1, and epoxide hydrolase. METHODS: chi(2) square analysis was used to compare frequencies of polymorphisms between 151 patients with Crohn's disease and 149 healthy controls. RESULTS: In patients, a genetic polymorphism in exon 3 of the microsomal epoxide hydrolase gene was distributed significantly different compared with controls (chi(2)=23.7; p<0.0001). All other polymorphisms tested were equally distributed between patients and controls. CONCLUSIONS: Microsomal epoxide hydrolase may play a role in the pathophysiology of Crohn's disease. Furthermore, the epoxide hydrolase gene is located on chromosome 1q, close to a region previously linked to Crohn's disease. PMID- 12631668 TI - Early development of stricturing or penetrating pattern in Crohn's disease is influenced by disease location, number of flares, and smoking but not by NOD2/CARD15 genotype. AB - BACKGROUND: Crohn's disease is a heterogeneous entity. Disease behaviour, characterised as stricturing, penetrating, or non-stricturing non-penetrating, is a clinically important phenotype as it is associated with complications and need for surgery. It has recently been showed that the behaviour of Crohn's disease changes over the course of the disease. AIM: To assess the association between rapid development of a penetrating or stricturing pattern of Crohn's disease and demographic and clinical characteristics as well as NOD2/CARD15 genotype. PATIENTS AND METHODS: A total of 163 patients with a firm diagnosis of Crohn's disease and who had non-penetrating non-stricturing disease at diagnosis were studied. Various demographic and clinical characteristics as well as antisaccharomyces cerevisiae antibody status and NOD2/CARD15 genotype were documented in these patients. These characteristics were compared in subgroups of patients according to evolution of disease behaviour five years after diagnosis. RESULTS: Five years after diagnosis there were 110 (67.5%) patients with non- structuring non-penetrating disease, 18 (11%) with stricturing disease, and 35 (21.5%) with penetrating disease. In multivariate analysis, only disease location and number of flares per year were significantly discriminant between the three subgroups (p=0.0009 and 0.0001, respectively). Ileal location of the disease was associated with a stricturing pattern while a high number of flares was associated with a penetrating pattern. Active smoking was also associated with a penetrating pattern compared with a non-stricturing non-penetrating pattern only. CONCLUSIONS: Early development of stricturing or penetrating behaviour in Crohn's disease is influenced by disease location, clinical activity of the disease, and smoking habit, but not by NOD2/CARD15 genotype. PMID- 12631669 TI - CARD15/NOD2 gene variants are associated with familially occurring and complicated forms of Crohn's disease. AB - BACKGROUND: Variants of the caspase activating recruitment domain 15/nucleotide oligomerisation domain 2 (CARD15/NOD2) gene have been associated with susceptibility to Crohn's disease (CD). AIM: Our aim was to evaluate the allele frequencies of the CARD15 variants R702W, G908R, and 1007fs in Finnish inflammatory bowel disease (IBD) patients and to search for possible associations between CARD15 variants and occurrence of familial forms of IBD or complicated forms of CD. PATIENTS AND METHODS: We investigated 198 sporadic CD patients, 46 probands with familial CD, 27 CD probands from mixed IBD families, 99 unrelated patients with ulcerative colitis (UC), and 300 control individuals for the occurrence of the CARD15 gene variants R702W, G908R, and 1007fs. RESULTS: In CD patients, the allele frequencies for the rare variants of these polymorphisms were 3.3%, 0.6%, and 4.8% (total 8.7%), and the corresponding frequencies in healthy controls were 1.8%, 0%, and 1.7% (total 3.5%) (8.7% v 3.5%; p<0.01). In UC patients allele frequencies were comparable with those in controls. The frequency of the 1007fs polymorphism variant allele was significantly higher among all CD patients than in controls (4.8% v 1.7%; p<0.01) but there was no significant difference in allele frequencies between the CD and UC groups. The 1007fs allele frequency was higher in familial CD than in non-familial cases with CD (10.9% v 3.5%; p<0.01). There were no significant differences in the allele frequencies of the R702W and G908R polymorphisms between CD patients, UC patients, and controls. We found that 15.5% of CD patients, 9.1% of UC patients, and 6.7% of controls carried at least one of the CARD15 variants. In CD patients carrying at least one of the three NOD2 variants, the ileum was affected more often than in non-carrier CD patients (90% v 73%; p<0.05), they had stricturing or penetrating disease more often than non-carriers (88% v 56%; p<0.01), and they had an increased need for bowel surgery. CONCLUSIONS: The frequency of NOD2 gene variants was lower in genetically homogenous Finns than in other populations. The 1007fs variant was associated with CD. The occurrence of CARD15 variants predicted ileal location as well as stricturing and penetrating forms of CD. PMID- 12631670 TI - Association study of PHOX2B as a candidate gene for Hirschsprung's disease. AB - BACKGROUND: Hirschsprung's disease (HSCR) is a congenital disorder characterised by an absence of ganglion cells in the nerve plexuses of the lower digestive tract. Manifestation of the disease has been linked to mutations in genes that encode the crucial signals for the development of the enteric nervous system-the RET and EDNRB signalling pathways. The Phox2b gene is involved in neurogenesis and regulates Ret expression in mice, in which disruption of the Phox2b results in a HSCR-like phenotype. AIMS: To investigate the contribution of PHOX2B to the HSCR phenotype. METHODS: Using polymerase chain reaction amplification and direct sequencing, we screened PHOX2B coding regions and intron/exon boundaries for mutations and polymorphisms in 91 patients with HSCR and 71 ethnically matched controls. Seventy five HSCR patients with no RET mutations were independently considered. Haplotype and genotype frequencies were compared using the standard case control statistic. RESULTS: Sequence analysis revealed three new polymorphisms: two novel single nucleotide polymorphisms (A-->G(1364); A- >C(2607)) and a 15 base pair deletion (DEL(2609)). Statistically significant differences were found for A-->G(1364). Genotypes comprising allele G were underrepresented in patients (19% v 36%; chi(2)=9.30; p=0.0095 and 22% v 36%; chi(2)=7.38; p=0.024 for patients with no RET mutations). Pairwise linkage disequilibrium (LD) analysis revealed no LD between physically close polymorphisms indicating a hot spot for recombination in exon 3. CONCLUSION: The PHOX2B A-->G(1364) polymorphism is associated with HSCR. Whether it directly contributes to disease susceptibility or represents a marker for a locus in LD with PHOX2B needs further investigation. Our findings are in accordance with the involvement of PHOX2B in the signalling pathways governing the development of enteric neurones. PMID- 12631671 TI - Intestinal complications after chemotherapy for patients with unresected primary colorectal cancer and synchronous metastases. AB - BACKGROUND: The role of palliative resection of the primary tumour in patients who present with metastatic colorectal cancer is unclear. AIMS: This study compared the incidence of major intestinal complications in such patients who received chemotherapy treatment with or without prior palliative resection of the primary tumour. PATIENTS: The incidence of intestinal obstruction, perforation, fistula formation, and gastrointestinal haemorrhage, and the requirement for abdominal radiotherapy in patients with metastatic colorectal cancer treated at a single institution over a 10 year period was determined. RESULTS: Eighty two patients received initial treatment with chemotherapy without resection of the primary tumour (unresected group) and 280 patients had undergone prior resection (resected group). In the unresected group, the incidence of peritonitis, fistula formation, and intestinal haemorrhage was 2.4% (95% confidence interval (CI) 0.3 8.5%), 3.7% (95% CI 0.8-10.3%), and 3.7% (95% CI 0.8-10.3%), respectively, and was not significantly different from the resected group. Intestinal obstruction affected 13.4% (95% CI 6.9-22.7%) of patients in the unresected group and 13.2% (95% CI 9.2-17.2%) of patients in the resected group. More patients in the unresected group required >/=3 blood transfusions (14.6% v 7.5%; p=0.048) and abdominal radiotherapy (18.3% v 9.6%; p=0.03) than the resected group. CONCLUSIONS: The incidence of major intestinal complications in patients with unresected colorectal cancer and synchronous metastases who receive initial treatment with chemotherapy is low. Chemotherapy may be successfully used as initial treatment for such patients with no increased risk of most major intestinal complications compared with patients who have undergone initial resection of the primary tumour. PMID- 12631672 TI - Measurement and determinants of the natural history of liver fibrosis in hepatitis C virus infection: a cross sectional and longitudinal study. AB - INTRODUCTION: The rate of development of liver fibrosis in hepatitis C virus (HCV) infection varies between individuals. This accounts for the variation in duration of progression to cirrhosis. The aims of this study were: (1) to determine whether fibrosis progresses linearly through the grading scales and (2) to identify factors which influence the rate of fibrosis. METHODS: HCV infected patients who had undergone at least one liver biopsy were identified. Biopsies were scored using the modified HAI (Ishak) and METAVIR systems, which were compared. Patients were treatment naive at first biopsy. Demographic features were examined for their relationship to fibrosis rate (defined as fibrosis stage/infection duration) using univariate and multivariate analysis. A subgroup of patients with two biopsies was examined to test the assumption that fibrosis progresses in a linear fashion. RESULTS: A total of 917 patients were included. Male sex (p<0.00001), older age at infection (p90% of the cases, using isolated protein subunits as target structures. If contacts between subunits within quaternary structures are included, the accuracy increases to 97%. Interactions beyond atom-atom contact range were not required to distinguish native structures from the decoys using this method. The contact scoring performed as well or better than existing statistical and physicochemical potentials and may be applied as an independent means of evaluating putative structural models. PMID- 12631703 TI - Type III secretion systems and bacterial flagella: insights into their function from structural similarities. AB - Type III secretion systems and bacterial flagella are broadly compared at the level of their genetic structure, morphology, regulation, and function, integrating structural information, to provide an overview of how they might function at a molecular level. PMID- 12631704 TI - Identification of genes responsible for osteoblast differentiation from human mesodermal progenitor cells. AB - Single human bone marrow-derived mesodermal progenitor cells (MPCs) differentiate into osteoblasts, chondrocytes, adipocytes, myocytes, and endothelial cells. To identify genes involved in the commitment of MPCs to osteoblasts we examined the expressed gene profile of undifferentiated MPCs and MPCs induced to the osteoblast lineage for 1-7 days by cDNA microarray analysis. As expected, growth factor, hormone, and signaling pathway genes known to be involved in osteogenesis were activated during differentiation. In addition, 41 transcription factors (TFs) were differentially expressed over time, including TFs with known roles in osteoblast differentiation and TFs not known to be involved in osteoblast differentiation. As the latter group of TFs coclustered with osteogenesis specific TFs, they may play a role in osteoblast differentiation. When we compared the gene expression profile of MPCs induced to differentiate to chondroblasts and osteoblasts, significant differences in the nature and/or timing of gene activation were seen. These studies indicate that in vitro differentiation cultures in which MPCs are induced to one of multiple cell fates should be very useful for defining signals important for lineage-specific differentiation. PMID- 12631705 TI - Generalized singular value decomposition for comparative analysis of genome-scale expression data sets of two different organisms. AB - We describe a comparative mathematical framework for two genome-scale expression data sets. This framework formulates expression as superposition of the effects of regulatory programs, biological processes, and experimental artifacts common to both data sets, as well as those that are exclusive to one data set or the other, by using generalized singular value decomposition. This framework enables comparative reconstruction and classification of the genes and arrays of both data sets. We illustrate this framework with a comparison of yeast and human cell cycle expression data sets. PMID- 12631706 TI - Loss of protooncogene c-Myc function impedes G1 phase progression both before and after the restriction point. AB - c-myc is an important protooncogene whose misregulation is believed to causally affect the development of numerous human cancers. c-myc null rat fibroblasts are viable but display a severe (two- to threefold) retardation of proliferation. The rates of RNA and protein synthesis are reduced by approximately the same factor, whereas cell size remains unaffected. We have performed a detailed kinetic cell cycle analysis of c-myc(-/-) cells by using several labeling and synchronization methods. The majority of cells (>90%) in asynchronous, exponential phase c-myc(-/ ) cultures cycle continuously with uniformly elongated cell cycles. Cell cycle elongation is due to a major lengthening of G(1) phase (four- to fivefold) and a more limited lengthening of G(2) phase (twofold), whereas S phase duration is largely unaffected. Progression from mitosis to the G1 restriction point and the subsequent progression from the restriction point into S phase are both drastically delayed. These results are best explained by a model in which c-Myc directly affects cell growth (accumulation of mass) and cell proliferation (the cell cycle machinery) by independent pathways. PMID- 12631707 TI - Sac3 is an mRNA export factor that localizes to cytoplasmic fibrils of nuclear pore complex. AB - In eukaryotes, mRNAs are transcribed in the nucleus and exported to the cytoplasm for translation to occur. Messenger RNAs complexed with proteins referred to as ribonucleoparticles are recognized for nuclear export in part by association with Mex67, a key Saccharomyces cerevisiae mRNA export factor and homolog of human TAP/NXF1. Mex67, along with its cofactor Mtr2, is thought to promote ribonucleoparticle translocation by interacting directly with components of the nuclear pore complex (NPC). Herein, we show that the nuclear pore-associated protein Sac3 functions in mRNA export. Using a mutant allele of MTR2 as a starting point, we have identified a mutation in SAC3 in a screen for synthetic lethal interactors. Loss of function of SAC3 causes a strong nuclear accumulation of mRNA and synthetic lethality with a number of mRNA export mutants. Furthermore, Sac3 can be coimmunoprecipitated with Mex67, Mtr2, and other factors involved in mRNA export. Immunoelectron microscopy analysis shows that Sac3 localizes exclusively to cytoplasmic fibrils of the NPC. Finally, Mex67 accumulates at the nuclear rim when SAC3 is mutated, suggesting that Sac3 functions in Mex67 translocation through the NPC. PMID- 12631708 TI - Isoprenylcysteine carboxyl methyltransferase activity modulates endothelial cell apoptosis. AB - Extracellular ATP, adenosine (Ado), and adenosine plus homocysteine (Ado/HC) cause apoptosis of cultured pulmonary artery endothelial cells through the enhanced formation of intracellular S-adenosylhomocysteine and disruption of focal adhesion complexes. Because an increased intracellular ratio of S adenosylhomocysteine/S-adenosylmethionine favors inhibition of methylation, we hypothesized that Ado/HC might act by inhibition of isoprenylcysteine-O-carboxyl methyltransferase (ICMT). We found that N-acetyl-S-geranylgeranyl-L-cysteine (AGGC) and N-acetyl-S-farnesyl-L-cysteine (AFC), which inhibit ICMT by competing with endogenous substrates for methylation, caused apoptosis. Transient overexpression of ICMT inhibited apoptosis caused by Ado/HC, UV light exposure, or tumor necrosis factor-alpha. Because the small GTPase, Ras, is a substrate for ICMT and may modulate apoptosis, we also hypothesized that inhibition of ICMT with Ado/HC or AGGC might cause endothelial apoptosis by altering Ras activation. We found that ICMT inhibition decreased Ras methylation and activity and the activation of the downstream signaling molecules Akt, ERK-1, and ERK-2. Furthermore, overexpression of wild-type or dominant active H-Ras blocked Ado/HC induced apoptosis. These findings suggest that inhibition of ICMT causes endothelial cell apoptosis by attenuation of Ras GTPase methylation and activation and its downstream antiapoptotic signaling pathway. PMID- 12631709 TI - Grb2 regulates internalization of EGF receptors through clathrin-coated pits. AB - The molecular mechanisms of clathrin-dependent internalization of epidermal growth factor receptor (EGFR) are not well understood and, in particular, the sequence motifs that mediate EGFR interactions with coated pits have not been mapped. We generated a panel of EGFR mutants and stably expressed these mutants in porcine aortic endothelial (PAE) cells. Interestingly, mutations of tyrosine phosphorylation sites 1068 and 1086 that interact with growth-factor-receptor binding protein Grb2 completely abolished receptor internalization in PAE cells. Quantitative analysis of colocalization of EGF-rhodamine conjugate and coated pits labeled with yellow-fluorescent-protein-tagged beta2 subunit of clathrin adaptor complex AP-2 revealed that EGFR mutants lacking Grb2 binding sites do not efficiently enter coated pits. The depletion of Grb2 from PAE as well as HeLa cells expressing endogenous EGFRs by RNA interference substantially reduced the rate of EGFR internalization through clathrin-dependent pathway, thus providing the direct evidence for the important role of Grb2 in this process. Overexpression of Grb2 mutants, in which the SH3 domains were either deleted or inactivated by point mutations, significantly inhibited EGFR internalization in both PAE and HeLa cells. These findings indicate that Grb2, in addition to its key function in signaling through Ras, has a major regulatory role at the initial steps of EGFR internalization through clathrin-coated pits. Furthermore, the EGFR mutant lacking Grb2 binding sites did not efficiently recruit c-Cbl and was not polyubiquitinated. The data are consistent with the model whereby Grb2 participates in EGFR internalization through the recruitment of Cbl to the receptor, thus allowing proper ubiquitylation of EGFR and/or associated proteins at the plasma membrane. PMID- 12631710 TI - Roles of NUDE and NUDF proteins of Aspergillus nidulans: insights from intracellular localization and overexpression effects. AB - The NUDF protein of the filamentous fungus Aspergillus nidulans functions in the cytoplasmic dynein pathway. It binds several proteins, including the NUDE protein. Green fluorescent protein-tagged NUDF and NUDA (dynein heavy chain) localize to linearly moving dashes ("comets") that coincide with microtubule ends. Herein, deletion of the nudE gene did not eliminate the comets of NUDF and NUDA, but affected the behavior of NUDA. Comets were also observed with the green fluorescent protein-tagged NUDE and its nonfunctional C-terminal domain. In addition, overexpressed NUDA and NUDE accumulated in specks that were either immobile or bounced randomly. Neither comets nor specks were observed with the functional N-terminal domain of NUDE, indicating that these structures are not essential for NUDE function. Furthermore, NUDF overproduction totally suppressed deletion of the nudE gene. This implies that the function of NUDE is secondary to that of NUDF. Unexpectedly, NUDF overproduction inhibited one conditional nudA mutant and all tested apsA mutants. An allele-specific interaction between the nudF and nudA genes is consistent with a direct interaction between NUDF and dynein heavy chain. Because APSA and its yeast homolog Num1p are cortical proteins, an interaction between the nudF and apsA genes suggests a role for NUDF at the cell cortex. PMID- 12631711 TI - KDEL and KKXX retrieval signals appended to the same reporter protein determine different trafficking between endoplasmic reticulum, intermediate compartment, and Golgi complex. AB - Many endoplasmic reticulum (ER) proteins maintain their residence by dynamic retrieval from downstream compartments of the secretory pathway. In previous work we compared the retrieval process mediated by the two signals, KKMP and KDEL, by appending them to the same neutral reporter protein, CD8, and found that the two signals determine a different steady-state localization of the reporter. CD8-K (the KDEL-bearing form) was restricted mainly to the ER, whereas CD8-E19 (the KKMP-bearing form) was distributed also to the intermediate compartment and Golgi complex. To investigate whether this different steady-state distribution reflects a difference in exit rates from the ER and/or in retrieval, we have now followed the first steps of export of the two constructs from the ER and their trafficking between ER and Golgi complex. Contrary to expectation, we find that CD8-K is efficiently recruited into transport vesicles, whereas CD8-E19 is not. Thus, the more restricted ER localization of CD8-K must be explained by a more efficient retrieval to the ER. Moreover, because most of ER resident CD8-K is not O glycosylated but almost all CD8-E19 is, the results suggest that CD8-K is retrieved from the intermediate compartment, before reaching the Golgi, where O glycosylation begins. These results illustrate how different retrieval signals determine different trafficking patterns and pose novel questions on the underlying molecular mechanisms. PMID- 12631712 TI - The two variants of oxysterol binding protein-related protein-1 display different tissue expression patterns, have different intracellular localization, and are functionally distinct. AB - Oxysterol binding protein (OSBP) homologs comprise a family of 12 proteins in humans (Jaworski et al., 2001; Lehto et al., 2001). Two variants of OSBP-related protein (ORP) 1 have been identified: a short one that consists of the carboxy terminal ligand binding domain only (ORP1S, 437 aa) and a longer N-terminally extended form (ORP1L, 950 aa) encompassing three ankyrin repeats and a pleckstrin homology domain (PHD). We now report that the two mRNAs show marked differences in tissue expression. ORP1S predominates in skeletal muscle and heart, whereas ORP1L is the most abundant form in brain and lung. On differentiation of primary human monocytes into macrophages, both ORP1S and ORP1L mRNAs were induced, the up regulation of ORP1L being >100-fold. The intracellular localization of the two ORP1 variants was found to be different. Whereas ORP1S is largely cytosolic, the ORP1L variant localizes to late endosomes. A significant amount of ORP1S but only little ORP1L was found in the nucleus. The ORP1L ankyrin repeat region (aa 1-237) was found to localize to late endosomes such as the full-length protein. This localization was even more pronounced for a fragment that additionally includes the PHD (aa 1-408). The amino-terminal region of ORP1L consisting of the ankyrin repeat and PHDs is therefore likely to be responsible for the targeting of ORP1L to late endosomes. Interestingly, overexpression of ORP1L was found to enhance the LXRalpha-mediated transactivation of a reporter gene, whereas ORP1S failed to influence this process. The results suggest that the two forms of ORP1 are functionally distinct and that ORP1L is involved in control of cellular lipid metabolism. PMID- 12631713 TI - Organization and dynamics of growing microtubule plus ends during early mitosis. AB - A stable cell line expressing EB1-green fluorescent protein was used to image growing microtubule plus ends at the G(2)/M transition. By late prophase growing ends no longer extend to the cell periphery and were not uniformly distributed around each centrosome. Growing ends were much more abundant in the area surrounding the nuclear envelope, and microtubules growing around the nucleus were 1.5 fold longer than those growing in the opposite direction. The growth of longer ends toward the nucleus did not result from a localized faster growth rate, because this rate was approximately 11 microm/min in all directions from the centrosome. Rather, microtubule ends growing toward the nucleus seemed stabilized by dynein/dynactin associated with the nuclear envelope. Injection of p50 into late prophase cells removed dynein from the nuclear envelope, reduced the density of growing ends near the nuclear envelope and resulted in a uniform distribution of growing ends from each centrosome. We suggest that the cell cycle dependent binding of dynein/dynactin to the nuclear envelope locally stabilizes growing microtubules. Both dynein and microtubules would then be in a position to participate in nuclear envelope breakdown, as described in recent studies. PMID- 12631714 TI - HIV-1 envelope proteins complete their folding into six-helix bundles immediately after fusion pore formation. AB - Fusion proteins of many viruses, including HIV-1 envelope protein (Env), fold into six-helix bundle structures. Fusion between individual Env-expressing cells and target cells was studied by fluorescence microscopy, and a temperature jump technique, to determine whether folding of Env into a bundle is complete by the time fusion pores have formed. Lowering temperature to 4 degrees C immediately after a pore opened halted pore growth, which quickly resumed when temperature was raised again. HIV gp41-derived peptides that inhibit bundle formation (C34 or N36) caused the cold-arrested pore to quickly and irreversibly close, demonstrating that bundle formation is not complete by the time a pore has formed. In contrast, lowering the temperature to an intermediate value also halted pore growth, but the pore was not closed by the bundle-inhibiting peptides, and it enlarged when temperature was again elevated. This latter result shows that bundle formation is definitely required for the fusion process, but surprisingly, some (if not all) bundle formation occurs after a pore has formed. It is concluded that an essential function of the bundle is to stabilize the pore against collapse and ensure its growth. PMID- 12631715 TI - Peroxisome biogenesis occurs in an unsynchronized manner in close association with the endoplasmic reticulum in temperature-sensitive Yarrowia lipolytica Pex3p mutants. AB - Pex3p is a peroxisomal integral membrane protein required early in peroxisome biogenesis, and Pex3p-deficient cells lack identifiable peroxisomes. Two temperature-sensitive pex3 mutant strains of the yeast Yarrowia lipolytica were made to investigate the role of Pex3p in the early stages of peroxisome biogenesis. In glucose medium at 16 degrees C, these mutants underwent de novo peroxisome biogenesis and exhibited early matrix protein sequestration into peroxisome-like structures found at the endoplasmic reticulum-rich periphery of cells or sometimes associated with nuclei. The de novo peroxisome biogenesis seemed unsynchronized, with peroxisomes occurring at different stages of development both within cells and between cells. Cells with peripheral nascent peroxisomes and cells with structures morphologically distinct from peroxisomes, such as semi/circular tubular structures that immunostained with antibodies to peroxisomal matrix proteins and to the endoplasmic reticulum-resident protein Kar2p, and that surrounded lipid droplets, were observed during up-regulation of peroxisome biogenesis in cells incubated in oleic acid medium at 16 degrees C. These structures were not detected in wild-type or Pex3p-deficient cells. Their role in peroxisome biogenesis remains unclear. Targeting of peroxisomal matrix proteins to these structures suggests that Pex3p directly or indirectly sequesters components of the peroxisome biogenesis machinery. Such a role is consistent with Pex3p overexpression producing cells with fewer, larger, and clustered peroxisomes. PMID- 12631716 TI - Global transcription analysis of Krebs tricarboxylic acid cycle mutants reveals an alternating pattern of gene expression and effects on hypoxic and oxidative genes. AB - To understand the many roles of the Krebs tricarboxylic acid (TCA) cycle in cell function, we used DNA microarrays to examine gene expression in response to TCA cycle dysfunction. mRNA was analyzed from yeast strains harboring defects in each of 15 genes that encode subunits of the eight TCA cycle enzymes. The expression of >400 genes changed at least threefold in response to TCA cycle dysfunction. Many genes displayed a common response to TCA cycle dysfunction indicative of a shift away from oxidative metabolism. Another set of genes displayed a pairwise, alternating pattern of expression in response to contiguous TCA cycle enzyme defects: expression was elevated in aconitase and isocitrate dehydrogenase mutants, diminished in alpha-ketoglutarate dehydrogenase and succinyl-CoA ligase mutants, elevated again in succinate dehydrogenase and fumarase mutants, and diminished again in malate dehydrogenase and citrate synthase mutants. This pattern correlated with previously defined TCA cycle growth-enhancing mutations and suggested a novel metabolic signaling pathway monitoring TCA cycle function. Expression of hypoxic/anaerobic genes was elevated in alpha-ketoglutarate dehydrogenase mutants, whereas expression of oxidative genes was diminished, consistent with a heme signaling defect caused by inadequate levels of the heme precursor, succinyl-CoA. These studies have revealed extensive responses to changes in TCA cycle function and have uncovered new and unexpected metabolic networks that are wired into the TCA cycle. PMID- 12631717 TI - GLUT4 recycles via a trans-Golgi network (TGN) subdomain enriched in Syntaxins 6 and 16 but not TGN38: involvement of an acidic targeting motif. AB - Insulin stimulates glucose transport in fat and muscle cells by triggering exocytosis of the glucose transporter GLUT4. To define the intracellular trafficking of GLUT4, we have studied the internalization of an epitope-tagged version of GLUT4 from the cell surface. GLUT4 rapidly traversed the endosomal system en route to a perinuclear location. This perinuclear GLUT4 compartment did not colocalize with endosomal markers (endosomal antigen 1 protein, transferrin) or TGN38, but showed significant overlap with the TGN target (t)-soluble N ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) Syntaxins 6 and 16. These results were confirmed by vesicle immunoisolation. Consistent with a role for Syntaxins 6 and 16 in GLUT4 trafficking we found that their expression was up-regulated significantly during adipocyte differentiation and insulin stimulated their movement to the cell surface. GLUT4 trafficking between endosomes and trans-Golgi network was regulated via an acidic targeting motif in the carboxy terminus of GLUT4, because a mutant lacking this motif was retained in endosomes. We conclude that GLUT4 is rapidly transported from the cell surface to a subdomain of the trans-Golgi network that is enriched in the t-SNAREs Syntaxins 6 and 16 and that an acidic targeting motif in the C-terminal tail of GLUT4 plays an important role in this process. PMID- 12631718 TI - Short telomeres induce a DNA damage response in Saccharomyces cerevisiae. AB - Telomerase-deficient Saccharomyces cerevisiae cells show a progressive decrease in telomere length. When grown for several days in log phase, the tlc1Delta cells initially display wild-type growth kinetics with subsequent loss of growth potential after which survivors are generated via RAD52-dependent homologous recombination. We found that chromosome loss in these telomerase-deficient cells only increased after a significant decline in growth potential of the culture. At earlier stages of growth, as the telomerase-deficient cells began to show loss of growth potential, the cells arrested in G2/M and showed RNR3 induction and Rad53p phosphorylation. These responses were dependent on RAD24 and MEC1, suggesting that short telomeres are recognized as DNA damage and signal G2/M arrest. PMID- 12631719 TI - Specification of actin filament function and molecular composition by tropomyosin isoforms. AB - The specific functions of greater than 40 vertebrate nonmuscle tropomyosins (Tms) are poorly understood. In this article we have tested the ability of two Tm isoforms, TmBr3 and the human homologue of Tm5 (hTM5(NM1)), to regulate actin filament function. We found that these Tms can differentially alter actin filament organization, cell size, and shape. hTm5(NM1) was able to recruit myosin II into stress fibers, which resulted in decreased lamellipodia and cellular migration. In contrast, TmBr3 transfection induced lamellipodial formation, increased cellular migration, and reduced stress fibers. Based on coimmunoprecipitation and colocalization studies, TmBr3 appeared to be associated with actin-depolymerizing factor/cofilin (ADF)-bound actin filaments. Additionally, the Tms can specifically regulate the incorporation of other Tms into actin filaments, suggesting that selective dimerization may also be involved in the control of actin filament organization. We conclude that Tm isoforms can be used to specify the functional properties and molecular composition of actin filaments and that spatial segregation of isoforms may lead to localized specialization of actin filament function. PMID- 12631720 TI - Characterization of a new gammaTuRC subunit with WD repeats. AB - The gamma-tubulin ring complex (gammaTuRC), consisting of multiple protein subunits, can nucleate microtubule assembly. Although many subunits of the gammaTuRC have been identified, a complete set remains to be defined in any organism. In addition, how the subunits interact with each other to assemble into gammaTuRC remains largely unknown. Here, we report the characterization of a novel gammaTuRC subunit, Drosophila gamma ring protein with WD repeats (Dgp71WD). With the exception of gamma-tubulin, Dgp71WD is the only gammaTuRC component identified to date that does not contain the grip motifs, which are signature sequences conserved in gammaTuRC components. By performing immunoprecipitations after pair-wise coexpression in Sf9 cells, we show that Dgp71WD directly interacts with the grip motif-containing gammaTuRC subunits, Dgrips84, 91, 128, and 163, suggesting that Dgp71WD may play a scaffolding role in gammaTuRC organization. We also show that Dgrips128 and 163, like Dgrips84 and 91, can interact directly with gamma-tubulin. Coexpression of any of these grip motif containing proteins with gamma-tubulin promotes gamma-tubulin binding to guanine nucleotide. In contrast, in the same assay Dgp71WD interacts with gamma-tubulin but does not facilitate nucleotide binding. PMID- 12631721 TI - Loss of caveolin-1 gene expression accelerates the development of dysplastic mammary lesions in tumor-prone transgenic mice. AB - Caveolin-1 is the principal structural component of caveolae microdomains, which represent a subcompartment of the plasma membrane. Several independent lines of evidence support the notion that caveolin-1 functions as a suppressor of cell transformation. For example, the human CAV-1 gene maps to a suspected tumor suppressor locus (D7S522/7q31.1) that is frequently deleted in a number of carcinomas, including breast cancers. In addition, up to 16% of human breast cancers harbor a dominant-negative mutation, P132L, in the CAV-1 gene. Despite these genetic associations, the tumor suppressor role of caveolin-1 still remains controversial. To directly assess the in vivo transformation suppressor activity of the caveolin-1 gene, we interbred Cav-1 (-/-) null mice with tumor-prone transgenic mice (MMTV-PyMT) that normally develop multifocal dysplastic lesions throughout the entire mammary tree. Herein, we show that loss of caveolin-1 gene expression dramatically accelerates the development of these multifocal dysplastic mammary lesions. At 3 wk of age, loss of caveolin-1 resulted in an approximately twofold increase in the number of lesions (foci per gland; 3.3 +/- 1.0 vs. 7.0 +/- 1.2) and an approximately five- to sixfold increase in the total area occupied by these lesions. Similar results were obtained at 4 wk of age. However, complete loss of caveolin-1 was required to accelerate the appearance of these dysplastic mammary lesions, because Cav-1 (+/-) heterozygous mice did not show any increases in foci development. We also show that loss of caveolin-1 increases the extent and the histological grade of these mammary lesions and facilitates the development of papillary projections in the mammary ducts. Finally, we demonstrate that cyclin D1 expression levels are dramatically elevated in Cav-1 (-/-) null mammary lesions, consistent with the accelerated appearance and growth of these dysplastic foci. This is the first in vivo demonstration that caveolin-1 can function as a transformation suppressor gene. PMID- 12631722 TI - Sequential entry of components of the gene expression machinery into daughter nuclei. AB - In eukaryotic cells, RNA polymerase II (RNA pol II) transcription and pre-mRNA processing are coordinated events. We have addressed how individual components of the transcription and pre-mRNA processing machinery are organized during mitosis and subsequently recruited into the newly formed daughter nuclei. Interestingly, localization studies of numerous RNA pol II transcription and pre-mRNA processing factors revealed a nonrandom and sequential entry of these factors into daughter nuclei after nuclear envelope/lamina formation. The initiation competent form of RNA pol II and general transcription factors appeared in the daughter nuclei simultaneously, but prior to pre-mRNA processing factors, whereas the elongation competent form of RNA pol II was detected even later. The differential entry of these factors rules out the possibility that they are transported as a unitary complex. Telophase nuclei were competent for transcription and pre-mRNA splicing concomitant with the initial entry of the respective factors. In addition, our results revealed a low turnover rate of transcription and pre-mRNA splicing factors during mitosis. We provide evidence to support a model in which the entry of the RNA pol II gene expression machinery into newly forming daughter nuclei is a staged and ordered process. PMID- 12631724 TI - Dynamin participates in focal extracellular matrix degradation by invasive cells. AB - The degradation of extracellular matrix (ECM) by matrix metalloproteases is crucial in physiological and pathological cell invasion alike. Degradation occurs at specific sites where invasive cells make contact with the ECM via specialized plasma membrane protrusions termed invadopodia. Herein, we show that the dynamin 2 (Dyn2), a GTPase implicated in the control of actin-driven cytoskeletal remodeling events and membrane transport, is necessary for focalized matrix degradation at invadopodia. Dynamin was inhibited by using two approaches: 1) expression of dominant negative GTPase-impaired or proline-rich domain-deleted Dyn2 mutants; and 2) inhibition of the dynamin regulator calcineurin by cyclosporin A. In both cases, the number and extension of ECM degradation foci were drastically reduced. To understand the site and mechanism of dynamin action, the cellular structures devoted to ECM degradation were analyzed by correlative confocal light-electron microscopy. Invadopodia were found to be organized into a previously undescribed ECM-degradation structure consisting of a large invagination of the ventral plasma membrane surface in close spatial relationship with the Golgi complex. Dyn2 seemed to be concentrated at invadopodia. PMID- 12631723 TI - Myo3A, one of two class III myosin genes expressed in vertebrate retina, is localized to the calycal processes of rod and cone photoreceptors and is expressed in the sacculus. AB - The striped bass has two retina-expressed class III myosin genes, each composed of a kinase, motor, and tail domain. We report the cloning, sequence analysis, and expression patterns of the long (Myo3A) and short (Myo3B) class III myosins, as well as cellular localization and biochemical characterization of the long isoform, Myo3A. Myo3A (209 kDa) is expressed in the retina, brain, testis, and sacculus, and Myo3B (155 kDa) is expressed in the retina, intestine, and testis. The tails of these two isoforms contain two highly conserved domains, 3THDI and 3THDII. Whereas Myo3B has three IQ motifs, Myo3A has nine IQ motifs, four in its neck and five in its tail domain. Myo3A localizes to actin filament bundles of photoreceptors and is concentrated in the calycal processes. An anti-Myo3A antibody decorates the actin cytoskeleton of rod inner/outer segments, and this labeling is reduced by the presence of ATP. The ATP-sensitive actin association is a feature characteristic of myosin motors. The numerous IQ motifs may play a structural or signaling role in the Myo3A, and its localization to calycal processes indicates that this myosin mediates a local function at this site in vertebrate photoreceptors. PMID- 12631725 TI - A dynamin-cortactin-Arp2/3 complex mediates actin reorganization in growth factor stimulated cells. AB - The mechanisms by which mammalian cells remodel the actin cytoskeleton in response to motogenic stimuli are complex and a topic of intense study. Dynamin 2 (Dyn2) is a large GTPase that interacts directly with several actin binding proteins, including cortactin. In this study, we demonstrate that Dyn2 and cortactin function to mediate dynamic remodeling of the actin cytoskeleton in response to stimulation with the motogenic growth factor platelet-derived growth factor. On stimulation, Dyn2 and cortactin coassemble into large, circular structures on the dorsal cell surface. These "waves" promote an active reorganization of actin filaments in the anterior cytoplasm and function to disassemble actin stress fibers. Importantly, inhibition of Dyn2 and cortactin function potently blocked the formation of waves and subsequent actin reorganization. These findings demonstrate that cortactin and Dyn2 function together in a supramolecular complex that assembles in response to growth factor stimulation and mediates the remodeling of actin to facilitate lamellipodial protrusion at the leading edge of migrating cells. PMID- 12631726 TI - Polarized distribution of IQGAP proteins in gastric parietal cells and their roles in regulated epithelial cell secretion. AB - Actin cytoskeleton plays an important role in the establishment of epithelial cell polarity. Cdc42, a member of Rho GTPase family, modulates actin dynamics via its regulators, such as IQGAP proteins. Gastric parietal cells are polarized epithelial cells in which regulated acid secretion occurs in the apical membrane upon stimulation. We have previously shown that actin isoforms are polarized to different membrane domains and that the integrity of the actin cytoskeleton is essential for acid secretion. Herein, we show that Cdc42 is preferentially distributed to the apical membrane of gastric parietal cells. In addition, we revealed that two Cdc42 regulators, IQGAP1 and IQGAP2, are present in gastric parietal cells. Interestingly, IQGAP2 is polarized to the apical membrane of the parietal cells, whereas IQGAP1 is mainly distributed to the basolateral membrane. An IQGAP peptide that competes with full-length IQGAP proteins for Cdc42-binding in vitro also inhibits acid secretion in streptolysin-O-permeabilized gastric glands. Furthermore, this peptide disrupts the association of IQGAP and Cdc42 with the apical actin cytoskeleton and prevents the apical membrane remodeling upon stimulation. We propose that IQGAP2 forms a link that associates Cdc42 with the apical cytoskeleton and thus allows for activation of polarized secretion in gastric parietal cells. PMID- 12631727 TI - The fission yeast spo14+ gene encoding a functional homologue of budding yeast Sec12 is required for the development of forespore membranes. AB - The Schizosaccharomyces pombe spo14-B221 mutant was originally isolated as a sporulation-deficient mutant. However, the spo14(+) gene is essential for cell viability and growth. spo14(+) is identical to the previously characterized stl1(+) gene encoding a putative homologue of Saccharomyces cerevisiae Sec12, which is essential for protein transport from the endoplasmic reticulum (ER) to the Golgi apparatus. In the spo14 mutant cells, ER-like membranes were accumulated beneath the plasma membrane and the ER/Golgi shuttling protein Rer1 remained in the ER. Sec12 is a guanine nucleotide exchange factor for the Sar1 GTPase. Overproduction of psr1(+) coding for an S. pombe Sar1 homologue suppressed both the sporulation defect of spo14-B221 and cold-sensitive growth of newly isolated spo14-6 and spo14-7 mutants. These results indicate that Spo14 is involved in early steps of the protein secretory pathway. The spo14-B221 allele carries a single nucleotide change in the branch point consensus of the fifth intron, which reduces the abundance of the spo14 mRNA. During meiosis II, the forespore membrane was initiated near spindle pole bodies; however, subsequent extension of the membrane was arrested before its closure into a sac. We conclude that Spo14 is responsible for the assembly of the forespore membrane by supplying membrane vesicles. PMID- 12631728 TI - Pre-M phase-promoting factor associates with annulate lamellae in Xenopus oocytes and egg extracts. AB - We have used complementary biochemical and in vivo approaches to study the compartmentalization of M phase-promoting factor (MPF) in prophase Xenopus eggs and oocytes. We first examined the distribution of MPF (Cdc2/CyclinB2) and membranous organelles in high-speed extracts of Xenopus eggs made during mitotic prophase. These extracts were found to lack mitochondria, Golgi membranes, and most endoplasmic reticulum (ER) but to contain the bulk of the pre-MPF pool. This pre-MPF could be pelleted by further centrifugation along with components necessary to activate it. On activation, Cdc2/CyclinB2 moved into the soluble fraction. Electron microscopy and Western blot analysis showed that the pre-MPF pellet contained a specific ER subdomain comprising "annulate lamellae" (AL): stacked ER membranes highly enriched in nuclear pores. Colocalization of pre-MPF with AL was demonstrated by anti-CyclinB2 immunofluorescence in prophase oocytes, in which AL are positioned close to the vegetal surface. Green fluorescent protein-CyclinB2 expressed in oocytes also localized at AL. These data suggest that inactive MPF associates with nuclear envelope components just before activation. This association may explain why nuclei and centrosomes stimulate MPF activation and provide a mechanism for targeting of MPF to some of its key substrates. PMID- 12631729 TI - Obscurin is a ligand for small ankyrin 1 in skeletal muscle. AB - The factors that organize the internal membranes of cells are still poorly understood. We have been addressing this question using striated muscle cells, which have regular arrays of membranes that associate with the contractile apparatus in stereotypic patterns. Here we examine links between contractile structures and the sarcoplasmic reticulum (SR) established by small ankyrin 1 (sAnk1), a approximately 17.5-kDa integral protein of network SR. We used yeast two-hybrid to identify obscurin, a giant Rho-GEF protein, as the major cytoplasmic ligand for sAnk1. The binding of obscurin to the cytoplasmic sequence of sAnk1 is mediated by a sequence of obscurin that is C-terminal to its last Ig like domain. Binding was confirmed in two in vitro assays. In one, GST-obscurin, bound to glutathione-matrix, specifically adsorbed native sAnk1 from muscle homogenates. In the second, MBP-obscurin bound recombinant GST-sAnk1 in nitrocellulose blots. Kinetic studies using surface plasmon resonance yielded a K(D) = 130 nM. On subcellular fractionation, obscurin was concentrated in the myofibrillar fraction, consistent with its identification as sarcomeric protein. Nevertheless, obscurin, like sAnk1, concentrated around Z-disks and M-lines of striated muscle. Our findings suggest that obscurin binds sAnk1, and are the first to document a specific and direct interaction between proteins of the sarcomere and the SR. PMID- 12631730 TI - Analysis of Na+,K+-ATPase motion and incorporation into the plasma membrane in response to G protein-coupled receptor signals in living cells. AB - Dopamine (DA) increases Na(+),K(+)-ATPase activity in lung alveolar epithelial cells. This effect is associated with an increase in Na(+),K(+)-ATPase molecules within the plasma membrane (). Analysis of Na(+),K(+)-ATPase motion was performed in real-time in alveolar cells stably expressing Na(+),K(+)-ATPase molecules carrying a fluorescent tag (green fluorescent protein) in the alpha-subunit. The data demonstrate a distinct (random walk) pattern of basal movement of Na(+),K(+) ATPase-containing vesicles in nontreated cells. DA increased the directional movement (by 3.5 fold) of the vesicles and an increase in their velocity (by 25%) that consequently promoted the incorporation of vesicles into the plasma membrane. The movement of Na(+),K(+)-ATPase-containing vesicles and incorporation into the plasma membrane were microtubule dependent, and disruption of this network perturbed vesicle motion toward the plasma membrane and prevented the increase in the Na(+),K(+)-ATPase activity induced by DA. Thus, recruitment of new Na(+),K(+)-ATPase molecules into the plasma membrane appears to be a major mechanism by which dopamine increases total cell Na(+),K(+)-ATPase activity. PMID- 12631731 TI - Hic-5 communicates between focal adhesions and the nucleus through oxidant sensitive nuclear export signal. AB - hic-5 was originally isolated as an H(2)O(2)-inducible cDNA clone whose product was normally found at focal adhesions. In this study, we found that Hic-5 accumulated in the nucleus in response to oxidants such as H(2)O(2). Other focal adhesion proteins including paxillin, the most homologous to Hic-5, remained in the cytoplasm. Mutation analyses revealed that the C- and N-terminal halves of Hic-5 contributed to its nuclear localization in a positive and negative manner, respectively. After the finding that leptomycin B (LMB), an inhibitor of nuclear export signal (NES), caused Hic-5 to be retained in the nucleus, Hic-5 was demonstrated to harbor NES in the N-terminal, which was sensitive to oxidants, thereby regulating the nuclear accumulation of Hic-5. NES consisted of a leucine rich stretch and two cysteines with a limited similarity to Yap/Pap-type NES. In the nucleus, Hic-5 was suggested to participate in the gene expression of c-fos. Using dominant negative mutants, we found that Hic-5 was actually involved in endogenous c-fos gene expression upon H(2)O(2) treatment. Hic-5 was thus proposed as a focal adhesion protein with the novel aspect of shuttling between focal adhesions and the nucleus through an oxidant-sensitive NES, mediating the redox signaling directly to the nucleus. PMID- 12631732 TI - Specificity of class II Hsp40 Sis1 in maintenance of yeast prion [RNQ+]. AB - Sis1 and Ydj1, functionally distinct heat shock protein (Hsp)40 molecular chaperones of the yeast cytosol, are homologs of Hdj1 and Hdj2 of mammalian cells, respectively. Sis1 is necessary for propagation of the Saccharomyces cerevisiae prion [RNQ(+)]; Ydj1 is not. The ability to function in [RNQ(+)] maintenance has been conserved, because Hdj1 can function to maintain Rnq1 in an aggregated form in place of Sis1, but Hdj2 cannot. An extended glycine-rich region of Sis1, composed of a region rich in phenylalanine residues (G/F) and another rich in methionine residues (G/M), is critical for prion maintenance. Single amino acid alterations in a short stretch of amino acids of the G/F region of Sis1 that are absent in the otherwise highly conserved G/F region of Ydj1 cause defects in prion maintenance. However, there is some functional redundancy within the glycine-rich regions of Sis1, because a deletion of the adjacent glycine/methionine (G/M) region was somewhat defective in propagation of [RNQ(+)] as well. These results are consistent with a model in which the glycine-rich regions of Hsp40s contain specific determinants of function manifested through interaction with Hsp70s. PMID- 12631733 TI - Essential roles for GPI-anchored proteins in African trypanosomes revealed using mutants deficient in GPI8. AB - The survival of Trypanosoma brucei, the causative agent of Sleeping Sickness and Nagana, is facilitated by the expression of a dense surface coat of glycosylphosphatidylinositol (GPI)-anchored proteins in both its mammalian and tsetse fly hosts. We have characterized T. brucei GPI8, the gene encoding the catalytic subunit of the GPI:protein transamidase complex that adds preformed GPI anchors onto nascent polypeptides. Deletion of GPI8 (to give Deltagpi8) resulted in the absence of GPI-anchored proteins from the cell surface of procyclic form trypanosomes and accumulation of a pool of non-protein-linked GPI molecules, some of which are surface located. Procyclic Deltagpi8, while viable in culture, were unable to establish infections in the tsetse midgut, confirming that GPI-anchored proteins are essential for insect-parasite interactions. Applying specific inducible GPI8 RNAi with bloodstream form parasites resulted in accumulation of unanchored variant surface glycoprotein and cell death with a defined multinuclear, multikinetoplast, and multiflagellar phenotype indicative of a block in cytokinesis. These data show that GPI-anchored proteins are essential for the viability of bloodstream form trypanosomes even in the absence of immune challenge and imply that GPI8 is important for proper cell cycle progression. PMID- 12631734 TI - Mutant proinsulin that cannot be converted is secreted efficiently from primary rat beta-cells via the regulated pathway. AB - Prohormones are directed from the trans-Golgi network to secretory granules of the regulated secretory pathway. It has further been proposed that prohormone conversion by endoproteolysis may be necessary for subsequent retention of peptides in granules and to prevent their release by the so-called "constitutive like" pathway. To address this directly, mutant human proinsulin (Arg/Gly(32):Lys/Thr(64)), which cannot be cleaved by conversion endoproteases, was expressed in primary rat islet cells by recombinant adenovirus. The handling of the mutant proinsulin was compared with that of wild-type human proinsulin. Infected islet cells were pulse labeled and both basal and stimulated secretion of radiolabeled products followed during a chase. Labeled products were quantified by high-performance liquid chromatography. As expected, the mutant proinsulin was not converted at any time. Basal (constitutive and constitutive like) secretion was higher for the mutant proinsulin than for wild-type proinsulin/insulin, but amounted to <1% even during a prolonged (6-h) period of basal chase. There was no difference in stimulated (regulated) secretion of mutant and wild-type proinsulin/insulin at any time. Thus, in primary islet cells, unprocessed (mutant) proinsulin is sorted to the regulated pathway and then retained in secretory granules as efficiently as fully processed insulin. PMID- 12631735 TI - Tor kinases are in distinct membrane-associated protein complexes in Saccharomyces cerevisiae. AB - Tor1p and Tor2p kinases, targets of the immune-suppressive antibiotic rapamycin, are components of a highly conserved signaling network that couples nutrient availability and cell growth. To gain insight into the molecular basis underlying Tor-dependent signaling, we used cell fractionation and immunoaffinity chromatography to examine the physical environment of Tor2p. We found that the majority of Tor2p associates with a membrane-bound compartment along with at least four other proteins, Avo1p-Avo3p and Lst8p. Using immunogold electron microscopy, we observed that Tor2p, as well as Tor1p, localizes in punctate clusters to regions adjacent to the plasma membrane and within the cell interior, often in association with characteristic membranous tracks. Cell fractionation, coimmunoprecipitation, and immunogold electron microscopy experiments confirmed that Lst8 associates with both Tor2p as well as Tor1p at these membranous sites. In contrast, we find that Kog1, the yeast homologue of the mammalian Tor regulatory protein Raptor, interacts preferentially with Tor1p. These findings provide evidence for the existence of Tor signaling complexes that contain distinct as well as overlapping components. That these complexes colocalize to a membrane-bound compartment suggests an intimate relationship between membrane mediated signaling and Tor activity. PMID- 12631736 TI - Importin-alpha mediates the regulated nuclear targeting of serum- and glucocorticoid-inducible protein kinase (Sgk) by recognition of a nuclear localization signal in the kinase central domain. AB - The transcriptionally regulated serum and glucocorticoid inducible protein kinase (Sgk) is localized to the nucleus in a serum-dependent manner, and a yeast two hybrid genetic screen uncovered a specific interaction between Sgk and the importin-alpha nuclear import receptor. In vitro GST pull down assays demonstrated a strong and direct association of importin-alpha with endogenous Sgk and exogenously expressed HA-tagged Sgk, whereas both components coimmunoprecipitate and colocalize to the nucleus after serum stimulation. Consistent with an active mechanism of nuclear localization, the nuclear import of HA-Sgk in permeabilized cells required ATP, cytoplasm, and a functional nuclear pore complex. Ectopic addition of a 107 amino acid carboxy-terminal fragment of importin-alpha, which contains the Sgk binding region, competitively inhibited the ability of endogenous importin-alpha to import Sgk into nuclei in vitro. Mutagenesis of lysines by alanine substitution defined a KKAILKKKEEK sequence within the central domain of Sgk between amino acids 131-141 that functions as a nuclear localization signal (NLS) required for the in vitro interaction with importin-alpha and for nuclear import of full-length Sgk in cultured cells. The serum-induced nuclear import of Sgk requires the NLS dependent recognition of Sgk by importin-alpha as well as the PI3-kinase dependent phosphorylation of Sgk. Our results define a new role importin-alpha in the stimulus-dependent control of signal transduction by nuclear localized protein kinases. PMID- 12631737 TI - Drs2p-related P-type ATPases Dnf1p and Dnf2p are required for phospholipid translocation across the yeast plasma membrane and serve a role in endocytosis. AB - Plasma membranes in eukaryotic cells display asymmetric lipid distributions with aminophospholipids concentrated in the inner and sphingolipids in the outer leaflet. This asymmetry is maintained by ATP-driven lipid transporters whose identities are unknown. The yeast plasma membrane contains two P-type ATPases, Dnf1p and Dnf2p, with structural similarity to ATPase II, a candidate aminophospholipid translocase from bovine chromaffin granules. Loss of Dnf1p and Dnf2p virtually abolished ATP-dependent transport of NBD-labeled phosphatidylethanolamine, phosphatidylserine, and phosphatidylcholine from the outer to the inner plasma membrane leaflet, leaving transport of sphingolipid analogs unaffected. Labeling with trinitrobenzene sulfonic acid revealed that the amount of phosphatidylethanolamine exposed on the surface of Deltadnf1Deltadnf2 cells increased twofold relative to wild-type cells. Phosphatidylethanolamine exposure by Deltadnf1Deltadnf2 cells further increased upon removal of Drs2p, an ATPase II homolog in the yeast Golgi. These changes in lipid topology were accompanied by a cold-sensitive defect in the uptake of markers for bulk-phase and receptor-mediated endocytosis. Our findings demonstrate a requirement for Dnf1p and Dnf2p in lipid translocation across the yeast plasma membrane. Moreover, it appears that Dnf1p, Dnf2p and Drs2p each help regulate the transbilayer lipid arrangement in the plasma membrane, and that this regulation is critical for budding endocytic vesicles. PMID- 12631738 TI - Subnuclear localization and Cajal body targeting of transcription elongation factor TFIIS in amphibian oocytes. AB - We have examined the localization and targeting of the RNA polymerase II (pol II) transcription elongation factor TFIIS in amphibian oocyte nuclei by immunofluorescence. Using a novel antibody against Xenopus TFIIS the major sites of immunostaining were found to be Cajal bodies, nuclear organelles that also contain pol II. Small granular structures attached to lampbrush chromosomes were also specifically stained but the transcriptionally active loops were not. Similar localization patterns were found for the newly synthesized myc-tagged TFIIS produced after injection of synthetic transcripts into the cytoplasm. The basis of the rapid and preferential targeting of TFIIS to Cajal bodies was investigated by examining the effects of deletion and site-specific mutations. Multiple regions of TFIIS contributed to efficient targeting including the domain required for its binding to pol II. The localization of TFIIS in Cajal bodies, and in particular the apparent involvement of pol II binding in achieving it, offer further support for a model in which Cajal bodies function in the preassembly of the transcriptional machinery. Although our findings are therefore consistent with TFIIS playing a role in early events of the transcription cycle, they also suggest that this elongation factor is not generally required during transcription in oocytes. PMID- 12631739 TI - Recognition of a single transmembrane degron by sequential quality control checkpoints. AB - To understand the relationship between conformational maturation and quality control-mediated proteolysis in the secretory pathway, we engineered the well characterized degron from the alpha-subunit of the T-cell antigen receptor (TCRalpha) into the alpha-helical transmembrane domain of homotrimeric type I integral membrane protein, influenza hemagglutinin (HA). Although the membrane degron does not appear to interfere with acquisition of native secondary structure, as assessed by the formation of native intrachain disulfide bonds, only approximately 50% of nascent mutant HA chains (HA(++)) become membrane integrated and acquire complex N-linked glycans indicative of transit to a post ER compartment. The remaining approximately 50% of nascent HA(++) chains fail to integrate into the lipid bilayer and are subject to proteasome-dependent degradation. Site-specific cleavage by extracellular trypsin and reactivity with conformation-specific monoclonal antibodies indicate that membrane-integrated HA(++) molecules are able to mature to the plasma membrane with a conformation indistinguishable from that of HA(wt). These apparently native HA(++) molecules are, nevertheless, rapidly degraded by a process that is insensitive to proteasome inhibitors but blocked by lysosomotropic amines. These data suggest the existence in the secretory pathway of at least two sequential quality control checkpoints that recognize the same transmembrane degron, thereby ensuring the fidelity of protein deployment to the plasma membrane. PMID- 12631741 TI - Outbreak of Salmonella dublin infection in France, November - December 1995. AB - On 20 December 1995, the National Network of Public Health (Reseau National de Sante Publique - RNSP) was notified by the Salmonella and Shigella National Reference Centre (Centre National de Reference - CNR) that a greater than expected number of human i PMID- 12631742 TI - An outbreak of cryptosporidiosis in the Netherlands. AB - A hospital microbiologist in Spijkenisse, in the south west of the Netherlands - who had recently attended a parasitology course - identified cryptosporidial oocysts in stools from a patient with diarrhoea on 16 August 1995. Re-examination of 89 stool spe PMID- 12631740 TI - Mechanism of a transcriptional cross talk between transforming growth factor-beta regulated Smad3 and Smad4 proteins and orphan nuclear receptor hepatocyte nuclear factor-4. AB - We have shown previously that the transforming growth factor-beta (TGFbeta) regulated Sma-Mad (Smad) protein 3 and Smad4 proteins transactivate the apolipoprotein C-III promoter in hepatic cells via a hormone response element that binds the nuclear receptor hepatocyte nuclear factor 4 (HNF-4). In the present study, we show that Smad3 and Smad4 but not Smad2 physically interact with HNF-4 via their Mad homology 1 domains both in vitro and in vivo. The synergistic transactivation of target promoters by Smads and HNF-4 was shown to depend on the specific promoter context and did not require an intact beta hairpin/DNA binding domain of the Smads. Using glutathione S-transferase interaction assays, we established that two regions of HNF-4, the N-terminal activation function 1 (AF-1) domain (aa 1-24) and the C-terminal F domain (aa 388 455) can mediate physical Smad3/HNF-4 interactions in vitro. In vivo, Smad3 and Smad4 proteins enhanced the transactivation function of various GAL4-HNF-4 fusion proteins via the AF-1 and the adjacent DNA binding domain, whereas a single tyrosine to alanine substitution in AF-1 abolished coactivation by Smads. The findings suggest that the transcriptional cross talk between the TGFbeta regulated Smads and HNF-4 is mediated by specific functional domains in the two types of transcription factors. Furthermore, the specificity of this interaction for certain target promoters may play an important role in various hepatocyte functions, which are regulated by TGFbeta and the Smads. PMID- 12631743 TI - Response to a suspected case of diphtheria in Dordrecht, the Netherlands. AB - About 50 of the refugees in Dordrecht are from the Newly Independent States (NIS) of the former USSR, where diphtheria is rife. Cases have arisen in western Europe through contact with cases from the NIS. No NIS-related cases of diphtheria have been seen PMID- 12631744 TI - Diphtheria s European come back. AB - Since children have been vaccinated routinely, diphtheria has been eliminated in most European states and the recent occurrence of a large epidemic in the Newly Independent States (NIS) of the former USSR was unexpected. The resurgence of diphtheria in th PMID- 12631745 TI - Increase of syphilis in Finland related to the Russian epidemic. AB - In the early 1990s only 30 to 40 new serologically confirmed cases of syphilis were reported annually in Finland. Typical syphilis patients were heterosexual men who acquired the infection abroad. Since 1993, the incidence of syphilis has increased. In 19 PMID- 12631746 TI - Outbreak of Escherichia coli O157 in Sweden. AB - The Swedish Institute for Infectious Disease Control (Smittskyddsinstitutet - SMI) received no more than three reports of enterohaemorrhagic E. coli O157 infections (EHEC) each year from 1988 to 1994. About half of these were due to E. coli O157. In July PMID- 12631747 TI - An outbreak of shigellosis in a child care institution in Queluz, Portugal - 1995. AB - The introduction of shigella into a child care centre carries a high risk of secondary spread from person to person within the centre. We report an outbreak of shigellosis in early 1995 that affected 99 children, 17 of their relations, and seven workers i PMID- 12631748 TI - Surveillance of tuberculosis in Europe. AB - This paper summarises recommendations for uniform reporting on tuberculosis cases made by a working group set up in 1994 following a meeting on tuberculosis control in low prevalence countries. The meeting had been organised jointly by the World Health Or PMID- 12631749 TI - Cholera in the Mediterranean: outbreak in Albania. AB - In the 1990s, an epidemic of cholera caused by Vibrio cholerae 01 El Tor has spread from northern Pakistan to the Mediterranean. In 1993 sporadic cases of cholera, and epidemic foci, were recorded in many countries of the southern part of Eastern Europe. PMID- 12631750 TI - Hantavirus infections in Finland. AB - Hantaviruses cause haemorrhagic fever with renal syndrome (HFRS) and hantaviruspulmonary syndrome (HPS), and occur throughout the world. The viruses aretransmitted to humans in aerosols of rodent secretions. About 200 000 people,mainly in Eurasia, develop PMID- 12631752 TI - Immunisation schedules in the countries of the European Union. AB - The WHO Regional Office for Europe organises meetings on immunisation programmes for national experts from all countries of the European Union (EU)and data on the incidence of diseases and immunisation coverage are regularly submitted to WHO. We have anal PMID- 12631751 TI - A psittacosis outbreak in Belgian customs officers. AB - In March 1994, health authorities in Antwerp, Belgium, were informed that a member of a team of 15 customs officers had developed psittacosis. The customs officer concerned had been admitted to hospital with pneumonia 10 days after exposure to parakeets t PMID- 12631753 TI - Salm-Net: a network for human salmonella surveillance in Europe. AB - Salm-Net aims to prevent human salmonellosis within the EU by strengthening international laboratory based human salmonella surveillance and creating an on line European database of compatible data available to all participants. Salm Netis funded by DG XI PMID- 12631754 TI - ESEN : a comparison of vaccination programmes Part one: diphtheria. AB - The aim of the European Sero-Epidemiology Network (ESEN) is to coordinate and harmonise the serological surveillance of immunity to vaccine preventable diseases in eight countries in Europe (Denmark, England, France, Germany, Italy, the Netherlands, Finla PMID- 12631755 TI - Shellfish consumption and awareness of risk of acquiring hepatitis A among Neapolitan families - Italy 1997. AB - In 1996, about 15% of all reported cases of hepatitis A (n=8651) in Italy were from the Campania region, with Naples reporting the region s highest incidence (34.9/100 000). Almost a half of all cases were between 15 and 24 years of age. In recent years, PMID- 12631756 TI - Epidemiology of Mycoplasma pneumoniae infections in Poland : 28 years of surveillance in Warsaw 1970-1997. AB - Mycoplasma pneumoniae is a common cause of lower respiratory tract disease in humans, particularly among older children, adolescents, and young adults. Infections are endemic in cities and epidemic increases are observed at intervals of 4 to 7 years. M. p PMID- 12631757 TI - Outbreak of trichinellosis in the Midi-Pyrenees region of France January - March 1998. AB - Two outbreaks of trichinellosis in the Tarn et Garonne, departement, France were reported by the departmental health autorities on 2 March 1998, to the Reseau National de Sante Publique (RNSP). An epidemiological investigation began on 3 March in order to PMID- 12631759 TI - History of trichinellosis outbreaks linked to horse meat consumption 1975-1998. AB - Trichinellosis is a parasitic disease transmitted to humans by consumption of raw or undercooked meat from animals contaminated by worms of the Trichinella genus. In nature, the life cycle involves most carnivores, omnivores, and numerous rodents. Domesti PMID- 12631758 TI - Human outbreak of trichinellosis associated with the consumption of horsemeat in Italy. AB - Trichinellosis is endemic among sylvatic mammals in Italy, though it causes only few infections in humans, usually due to the consumption of pork from pigs grazing in wild areas or from wild boars. Most cases of human trichinellosis in Italy are due to th PMID- 12631760 TI - Trichinellosis associated with the consumption of horse meat: European regulations and risk management. AB - Outbreaks of trichinellosis in Italy and France at the end of 1975 implicated the horse, an animal not considered previously to be at risk of this disease. European regulations at that time concerned only the control of pork. The obligation to search for PMID- 12631761 TI - Viral marker rates among unpaid blood donors in Europe decreased from 1990 to 1996. AB - Not for profit plasma fractionators in Europe believe that the standardised measurement and reporting of such epidemiological data is a key issue for public health. Accordingly, under the auspices of the European Plasma Fractionation Association (EPFA), t PMID- 12631762 TI - Screening of viral markers for HIV HBV and HCV infections in blood donors in France and residual risk of viral transmission by blood transfusion. AB - The first part of this article presents the results of screening tests for antibodies to human immunodeficiency virus (HIV) and hepatitis C virus (HCV) and for hepatitis B surface antigen (HBsAg) from 1986 to 1996. The second part presents the most recent PMID- 12631763 TI - Establishing a European network for the diagnosis of. AB - The epidemics in recent years of Ebola haemorrhagic fever in Zaire and Gabon acted as a reminder that dangerous infections can be imported very quickly into Europe. Meetings on emerging and re-emerging pathogens organised by the World Health Organization PMID- 12631764 TI - Sentinel surveillance of sexually transmitted diseases in Italy. AB - Statutory notifications suggest that the incidence of gonorrhoea and syphilis have fallen in the past 30 years in Italy, as in other developed countries. Nevertheless, during the 1980s, the annual notification rates in Italy were from 15 to 50 times lower PMID- 12631765 TI - Surveillance of sexually transmitted diseases by laboratory networks in France. AB - In France, most of sexually transmitted diseases (STDs) are diagnosed by clinicians in the private sector (general practitioners, gynaecologists, dermatologists who specialise in venereal disease, and urologists) but genitourinary medicine (GUM) clinics a PMID- 12631766 TI - Surveillance of sexually transmitted infections in England and Wales. AB - Owing to the unique link between the epidemiology of sexually transmitted infections (STIs) and human behaviour, the surveillance of STIs requires the collection of more demographic and behavioural data than for other infectious diseases. In order to supp PMID- 12631767 TI - Surveillance of sexually transmitted infections in Scotland. AB - Applications of the surveillance of sexually transmitted infections (STIs) include setting priorities for and planning preventive and treatment services, evaluating interventions, and shaping research hypotheses. In Scotland routine data on STIs, (other t PMID- 12631768 TI - Surveillance of malaria in European Union countries. AB - The European Union countries are outside the endemic area for malaria, but many cases of malaria contracted elsewhere are imported into Europe each year. Several countries have reported high and increasing numbers of imported cases in recent years (France PMID- 12631770 TI - European Union inventory of resources and means of control of communicable diseases. AB - The Istituto Superiore di Sanita (ISS) in Rome, in collaboration with the Public Health Laboratory Service (PHLS) in London, and the Swedish Institute for Infectious Diseases Control (SIIDC) in Stockholm, is conducting a scientific and technical analysis PMID- 12631769 TI - Underreporting of legionnaires disease in France : the case for more active surveillance. AB - In France, surveillance of legionnaires disease started in 1987 with a mandatory clinician-based notification system. An average of 54 cases were reported yearly between 1988 and 1995 and six non-nosocomial clusters were identified. In 1995 the surveilla PMID- 12631771 TI - Recommendations for HIV surveillance in Europe. AB - The HIV/AIDS epidemic in Europe is now at an important turning point. In western Europe, AIDS incidence has been declining since 1996, partly as a result of new, more effective treatment of HIV infections. As these treatments delay the progression of HIV PMID- 12631772 TI - Imported malaria in Germany in 1996. AB - Malaria is not endemic in Germany, but it is one of the most commonly imported infectious diseases. The numbers of imported cases increased from 1994 to 1996; in 1997 according to preliminary data there were a total of 994 cases. In 1996, 1021 cases of ma PMID- 12631773 TI - Imported malaria in continental France in 1996. AB - Malaria ceased to be endemic in France in 1960, although sporadic cases were reported until 1970 in the department of Corse. No indigenous case of malaria have been reported since then. We report below the epidemiological data on imported malaria in 1996. PMID- 12631774 TI - Malaria surveillance in Italy : 1986-1996 analysis and 1997 provisional data. AB - A surveillance system for malaria was established in Italy to prevent a possible return of disease transmission after the eradication. Reporting malaria and 43 other infectious diseases is mandatory. Local laboratories diagnose clinical cases of malaria m PMID- 12631776 TI - The WHO update on influenza A (H5N1) in Hong Kong. AB - In May 1997 influenza A subtype H5N1 virus was isolated in the Special Administrative Region of Hong Kong from a child who died. Until then, the H5N1virus was known to infect only various species of birds, including chickens and ducks. After the first hum PMID- 12631775 TI - Malaria imported into the United Kingdom in 1996. AB - Virtually all malaria infections seen in the United Kingdom (UK) in recent years have been contracted overseas and imported into the UK, apart from two cases in 1976 due to imported infected mosquitoes and occasional cases of congenital or of transfusion PMID- 12631777 TI - The European Union faces up to the threat of a pandemic: meeting at the DGV on the influenza A (H5N1) of the ad hoc group on communicable diseases Luxembourg 14 January 1998. AB - The transmission of an avian influenza H5N1 virus to a child in Hong Kong in May 1997 followed by the occurrence of 17 other human cases with a high case fatality rate (6/18, 33%) suggested that an influenza pandemic could be imminent and led many Europea PMID- 12631778 TI - Plans against influenza pandemics in Europe: history and principles. AB - Antigenic variation, New Jersey porcine influenza, Pandemic planning: Berlin 1993, Pandemics, Principles of action, H5N1 avian influenza in Hong Kong. PMID- 12631779 TI - Sentinel surveillance of influenza in Europe 1997-1998. AB - The circulation of influenza virus is associated with increased consultation rates with general practitioners (GPs), increased hospital admissions, and excess deaths and has to be considered in health care planning. In western Europe, influenza viruses no PMID- 12631780 TI - Monitoring the introduction of universal influenza vaccination for the elderly people in the Netherlands. AB - For many years, influenza vaccination in the Netherlands has been administered by general practitioners (GPs), with whom every person is registered. Nine out of ten practices use one of six approved general practice information systems (GPIS). The exponen PMID- 12631781 TI - Surveillance of systemic invasive disease caused by group A Streptococcus in Italy 1994-1996. AB - In late May 1994, following reports of an outbreak of group A beta-haemolytic streptococcus (GAS) in the United Kingdom, the Istituto Superiore di Sanita (ISS) set up a surveillance system for systemic invasive GAS disease. The surveillance system was als PMID- 12631782 TI - An outbreak of mumps in the Province of Leon Spain 1995-1996. AB - In Spain mumps vaccine is given at the age of 15 months together with measles and rubella vaccines since 1982. Increased numbers of cases and outbreaks of mumps appeared in several autonomous communities in 1995. An outbreak of mumps in the province of - PMID- 12631783 TI - Is murine typhus re-emerging in Portugal ? AB - Murine typhus or endemic typhus is an infectious disease, clinically very similar to epidemic typhus, and caused by Rickettsia typhi(sometimes referred to as R. mooseri). Murine typhus was fairly common in Portugal until the 1940s, when several cases diag PMID- 12631786 TI - Surveillance of enterohaemorrhagic E. coli (EHEC) infections and haemolytic uraemic syndrome (HUS) in Europe. AB - Since they were first described, Escherichia coliO157: H7 and other related enterohaemorrhagic E. coli(EHEC) have become known as a major infectious cause of bloody diarrhoea. These E. coliproduce one or more shiga-toxins (stx) or Vero cytotoxins. Strictl PMID- 12631785 TI - European surveillance of travel associated legionnaires' disease 1996. AB - The European Surveillance Scheme for Travel Associated Legionnaires' Disease was established in 1987 to identify clusters and outbreaks of cases of the disease. Twenty-nine collaborating centres in 25 countries contribute case reports in a standard format PMID- 12631787 TI - Methicillin resistant Staphylococcus aureus (MRSA) as a community strain. AB - Methicillin resistant Staphylococcus aureus (MRSA) has become particularly well known in association with hospital acquired infections but is also known to have infected people in the community. We define cases of community acquired MRSA infection as the PMID- 12631788 TI - Increase of the number of vancomycin resistant enterococci (VRE) isolated in Helsinki, Finland. AB - The emergence of hospital acquired infections with bacteria resistant to antimicrobials such as vancomycin resistant enterococci (VRE) has become a worldwide concern. In hospitals in the United States, VRE have spread quickly and currently account for eve PMID- 12631790 TI - An outbreak of Salmonella enteritidis food poisoning in a psychiatric hospital in Dublin, Ireland. AB - On 29 August 1996 Ireland's Eastern Health Board (EHB) was informed of an outbreak of gastrointestinal illness in a psychiatric hospital in Dublin. Fifty people among 240 members of staff and 183 patients had reportedly fallen ill since 27 August and new PMID- 12631789 TI - Increasing incidence of resistance to trimethoprim and ciprofloxacin in epidemic Salmonella typhimurium DT104 in England and Wales. AB - Salmonella typhimurium definitive phage type (DT) 104 (= DT104) with chromosomally-encoded resistance to ampicillin, chloramphenicol, streptomycin, sulphonamides, and tetracyclines (= R-type ACSSuT) has become increasingly common in humans in England and PMID- 12631791 TI - Minimising the risk of salmonellosis from eggs. AB - Outbreaks of Salmonella enteritidis associated with raw eggs continue to be common, despite the risk of using raw eggs being well known. In many cases S. enteritidis isaself-limitingillnessbut- particularlyinvery young and very old people - it can be seve PMID- 12631793 TI - Recent changes in meningococcal disease in Europe. AB - Few infections can cause the social stress that occurs when meningococcal disease enters a community. The ability of Neisseria meningitidisto kill a healthy child within a few hours is one reason for this fear. It is also clear that preventive measures ag PMID- 12631792 TI - Measles in Germany. AB - Experts met in Copenhagen on 19 and 20 November 1996 to discuss the elimination of measles in Europe. European countries were assigned to four different categories on the basis of a recent survey conducted by the World Health Organization's Regional Offic PMID- 12631794 TI - Changing epidemiology of meningococcal disease in Spain, 1989-1997. AB - In the 1980s, the presentation of meningococcal disease in Spain was closely linked to the predominance of serotype B. Since the early 1990s however, an increasing number of serotype C strains have been isolated from a number of localities in the west of PMID- 12631795 TI - The epidemiology of meningococcal disease in England and Wales, 1996 and 1997. AB - Surveillance of meningococcal infection in England and Wales relies on three sources of data. Doctors are required by statute to notify clinically diagnosed cases of meningococcal meningitis and, since 1989, cases of meningococcal septicaemia (in the abse PMID- 12631796 TI - Trends in serogroup C meningococcal disease in the Republic of Ireland. AB - A national laboratory surveillance system for meningococcal disease was established in the Republic of Ireland (ROI) in November 1994. Data collection forms are returned centrally each month whether or not cases have occurred, containing a minimum data se PMID- 12631797 TI - Meningococcal infection in Scotland, 1995 to 1997. AB - The incidence of meningococcal infection in Scotland was 4.1 cases per 100 000 population in 1996 compared with 3.8/100 000 in 1995 (210 cases reported in 1996 compared with 193 in 1995). There were 14 deaths, representing a crude case fatality rate of 6. PMID- 12631799 TI - Surveillance of meningococcal disease in France, 1990-1997. AB - The surveillance of meningococcal disease in France is based on four complementary sources of data. 1. Meningococcal disease has been a notifiable disease since 1902. A case definition for surveillance was introduced in 1985, based on the isolation of Ne PMID- 12631798 TI - Cases of Neisseria meningitidis in Greece. AB - In Greece, Neisseria meningitidis appears as sporadic cases with some seasonal local outbreaks. The notification of meningococcal disease to the local health authorities and the Ministry of Health is mandatory. In 1994, the National Reference Centre for M PMID- 12631800 TI - New epidemiological features of meningococcal disease in Belgium. AB - SCIENTIFIC EDITOR In Belgium, after the epidemic of the early 1970s caused by Neisseria meningitidis B:2b:P1.2 (highest incidences in 1971 and 1972 with 5 cases per 100 000 population per year), the incidence of meningococcal disease fell to normal inter PMID- 12631801 TI - Why do adults contract diphtheria? AB - When diphtheria was a common disease, it most commonly affected children. Typically, at least 40% of diphtheria cases were children under 5 years of age, and some 70% of cases were children under 15 years of age. This classical pattern of diphtheria cases PMID- 12631802 TI - Diphtheria cases notified in the European Union. AB - Following the receipt of the articles on diphtheria from Poland published in this issue of EuroSurveillance, members of the editorial board were asked about the occurrence of diphtheria cases in their country in the last ten years (mid 1987 to mid 1997), PMID- 12631803 TI - Changes in age specific immunity to diphtheria in Poland in the past 40 years. AB - In recent years people in Poland have acquired diphtheria from eastern neighbours. It was considered important therefore to study the age specific immunity against diphtheria and to compare it with the results of seroepidemiological studies conducted in t PMID- 12631804 TI - A case report of laboratory-acquired diphtheria. AB - In March 1996, when a 41 year old laboratory technician developed malaise, low grade fever, sore throat, hoarseness and dysphagia she was initially treated symptomatically. One day later, the patient was seen in the emergency room of an ear, nose, and thr PMID- 12631805 TI - Eurosurveillance Weekly goes live. AB - Eurosurveillance Weekly, a weekly European bulletin on communicable diseases, and sister publication of Eurosurveillance, was launched on the world wide web in June. The Eurosurveillance Weekly homepage is updated once a week, but the frequency may be inc PMID- 12631806 TI - Development of a European electronic network on communicable diseases: the IDA HSSCD programme. AB - The European Commission (EC) has stated that setting up disease surveillance and health monitoring systems that cover the EU is a priority, and has proposed the development of a Community wide network, relying on electronic, computer based communications PMID- 12631807 TI - Changing patterns in acute clinical viral hepatitis encountered by Belgian general practitioners. PMID- 12631808 TI - Six years surveillance of hepatitis A and B in general practice in France. AB - The French sentinel network is composed of general practitioners who record on an ongoing basis cases of seven infectious diseases (i.e. influenza-like illness, acute diarrhoea, chickenpox, mumps, measles, male urethritis, and acute hepatitis of probable PMID- 12631809 TI - Outbreak of measles in adults in the north west of Greece. AB - The overall incidence of measles fell markedly in Greece after measles vaccination was introduced (although precise data are not available), but sporadic outbreaks continue to occur. A live monovalent measles vaccine was introduced in Greece in 1965 and r PMID- 12631810 TI - Travel associated legionellosis among European tourists in Spain. AB - Depuis la deuxieme moitie des annees 1980, la legionellose associee au voyage est devenue preoccupante pour les pays europeens en raison de la morbidite dans la population europeenne; de plus, l'apparition de cas chez les touristes constitue une menace p PMID- 12631811 TI - Travel associated legionellosis among European tourists in Spain - a comment from the EWGLI coordinating centre. AB - This interesting paper has analysed the data on cases of legionellosis associated with travel to Spain and highlighted some issues which are relevant to the European Working Group for Legionella Infections (EWGLI) surveillance system, eg the reporting of PMID- 12631812 TI - Outbreak of legionnaire s disease in two groups of tourists staying at camp sites in France. AB - On 11 June 1996, three suspected cases of legionnaires disease in a group of 42 Dutch tourists were reported to the local public health authority by Millau hospital in south west France. The group (group 1) had been touring with caravans and staying at d PMID- 12631813 TI - Human monkeypox in Kasai Oriental, Zaire (1996-1997). AB - Monkeypox is an orthopoxvirus with enzootic circulation in the rainforests of central and western Africa; the virus can be transmitted to humans and cause a syndrome clinically similar to smallpox (e.g., pustular rash, fever, respiratory symptoms, and in PMID- 12631814 TI - AIDS trends in Europe: decrease in the west, increase in the east. AB - In 1996, 24 920 AIDS cases were reported in the WHO European Region, bringing the cumulative total to 185 808 cases including 6969 (4%) paediatric cases (<13 years). For the first time since the start of the epidemic, the annual number of cases reported d PMID- 12631816 TI - Progress in Europe towards the goal of poliomyelitis eradication. AB - In 1988, the World Health Assembly set a target of global eradication of poliomyelitis due to wild poliovirus by the year 2000. The strategies of the campaign are: reach and maintain high levels of routine vaccination coverage throughout the population; p PMID- 12631815 TI - Polio outbreak in Albania, 1996. AB - Between April and November 1996, a large outbreak of polio occurred in Albania, which had reported to be free of polio since 1985. Although Albania had not reported polio in that interval, the risk of introduction and circulation of wild poliovirus had in PMID- 12631817 TI - Monitoring antimicrobial resistance in humans and animals in Europe. AB - Since publishing the editorial note on antimicrobial resistance ( Eurosurveillance 1997; 2(3): 21-2) we have received replies from Austria, Belgium, Greece, and Italy. Austria does not monitor antimicrobial resistance. The National Institute for Veterinar PMID- 12631818 TI - Widespread dissemination of epidemic MRSA in German hospitals. AB - Strains of Staphylococcus aureusresistant to multiple antibiotics, including those resistant to methicillin (MRSA), present a major problem in the control of hospital acquired infections. Some strains show a definite capacity for spread once introduced in PMID- 12631819 TI - Staphylococcus aureus and MRSA in Austrian hospitals, 1995. AB - In 1996, 327 acute care hospitals and other health care institutions in Austria were asked by the Federal Ministry of Health and Consumer Protection to report the number of inpatient episodes in 1995 in which Staphylococcus aureus (SA) was cultured. The m PMID- 12631820 TI - The European Sero-Epidemiology Network. AB - The purpose of serological surveillance is to monitor levels of immunity to particular diseases within a population in order to enhance the quality of information on which public health measures, such as vaccination programmes, are based. Serological surv PMID- 12631821 TI - A persisting outbreak of hepatitis A in Puglia, Italy, 1996: epidemiological follow-up. AB - In early 1996, an increase in the notifications of hepatitis A cases was identified in the region of Puglia, in the south east of Italy. An outbreak investigation conducted in May 1996 showed a strong association between illness and consumption of raw sea PMID- 12631822 TI - Surveillance of antimicrobial resistance in humans, food stuffs and livestock in Denmark. AB - A general increase in antimicrobial resistance among pathogenic bacteria is causing concern worldwide that the widespread use of antimicrobial agents in animal production may promote the development of resistant bacteria or resistance genes that can be tr PMID- 12631824 TI - Monitoring antimicrobial resistance in humans and animals in Europe. AB - Concern about possible transfer of bacterial resistance to antibiotics from animals to humans is growing at an international level. This is illustrated by the ban of avoparcin use as a growth promoter in livestock which will take effect in the European Un PMID- 12631823 TI - Surveillance of antibiotic resistance in Salmonella. AB - The World Health Organisation has recently pointed out an alarming increase in the incidence of antibiotic resistant strains of Salmonella, which are due to the use of antibiotics in intensive breeding. In France, until recent years, no or few cases of a PMID- 12631825 TI - Preliminary report of an international outbreak of Salmonella anatum infection linked to an infant formula milk. AB - The number of isolates of Salmonella anatum from infants (aged 1 to 11 months) in England and Wales was higher than expected in November and December 1996 and early January. The Public Health Laboratory Service (PHLS) Laboratory of Enteric Pathogens (LEP) PMID- 12631826 TI - Outbreak of Q fever in Lohra-Rollshausen, Germany, spring 1996. AB - Q fever is an acute (and sometimes chronic) febrile illness caused by the rickettsial organism Coxiella burnetii. The commonest animal reservoirs for C. burnetiiare cattle, sheep, and goats. Infected animals shed the organisms, which resist desiccation, i PMID- 12631828 TI - Q fever in Europe. AB - In order to give a European overview, members of the editorial board of Eurosurveillancewere asked a few questions about the surveillance of Q fever in the countries they represent and the possible occurrence of similar outbreaks in recent years. We rece PMID- 12631827 TI - Urban outbreak of Q fever, Briancon, France, March to June 1996. AB - Q fever is an ubiquitous zoonosis caused by the rickettsial organism Coxiella burnetii. Both sporadic cases and epidemics occur in areas where sheep and goats are bred. The main route of transmission is by inhalation of aerosols from the environment (soil PMID- 12631829 TI - Rotavirus in Ireland. AB - Acute diarrhoeal disease is the commonest single cause of morbidity and mortality worldwide. Infectious diarrhoea has been estimated to cause at least 5 million deaths each year in the developing world. Very young children are particularly susceptible to PMID- 12631830 TI - Scottish outbreak of Escherichia coli O157, November-December 1996. AB - On the afternoon of Friday 22 November 1996, Lanarkshire Health Board s Public Health Department became aware of 15 cases of possible Escherichia coli O157 infection (five of which had been confirmed microbiologically) in residents of the town of Wishaw i PMID- 12631831 TI - Immunisation coverage in the European Union. AB - The 15 countries of the European Union have ratified the fifth objective of Health for all in the year 2000 programme of WHO Europe, which states: By the year 2000, there should be no indigenous cases of poliomyelitis, diphtheria, neonatal tetanus, measl PMID- 12631832 TI - Salmonella enteritidis and S. typhimurium in Western Europe for 1993-1995: a surveillance report from Salm-Net. AB - Surveillance has been defined as the ongoing systematic collection, collation and analysis of data and the prompt dissemination of the resulting information to those who need to know so that an action can result. This is achieved in two ways. Firstly, i PMID- 12631833 TI - Outbreak of Salmonella enteritidis phage type 1 infection in British tourists visiting Mallorca, June 1996. AB - Three British tourists became ill while on holiday in the same hotel in Alcudia, Mallorca from 11 to 25 June 1996; Salmonella enteritidisinfections were diagnosed upon their return to England. An environmental health officer informed the Gastrointestinal PMID- 12631835 TI - Bacterial meningitis in Europe for 1994. AB - Thirty-five European countries contributed surveillance data to the fourth annual report of the King's European Meningococcal Surveillance Unit in London, which gathers and shares information about the epidemiology of meningococcal disease to inform publi PMID- 12631834 TI - Creutzfeldt-Jakob disease: results of an inquiry in the fifteen Member States of the European Union. AB - Creutzfeldt-Jakob Disease (CJD) was first described in 1920-1921. CJD is a rare disease with a reported incidence of 0.5 to 1 case per million people in Europe. This fatal dementia belongs to the category of Transmissible Spongiform Encephalopathies (TSE) PMID- 12631836 TI - Editorial. AB - Over 30 years ago, AD Langmuir defined surveillance as the ongoing systematic collection, collation and analysis of data and the prompt dissemination of the resulting information to those who need to know so that an action can result. The countries of t PMID- 12631837 TI - Outbreak of measles among adolescents, Belgium, 1996. AB - Measles was very common in Belgium before 1985, affecting 95% of children before they reached the age of 15 years. In 1984 the incidence of measles, calculated using data from a general practitioner surveillance system, was 823/100 000 inhabitants (2). Wi PMID- 12631838 TI - An outbreak of hepatitis A in Puglia, Italy, 1996. AB - In the region of Puglia, in the south east of Italy (population: 4 million), the number of notifications of hepatitis A virus (HAV) infection increases in a two yearly cycle. In 1992 a very large outbreak was observed, but no epidemiological investigation PMID- 12631839 TI - Value of influenza vaccine during an outbreak of influenza A in a nursing home, Pyrenees Atlantiques, France, November-December 1995. AB - A public health officer of the Direction Departementale de l'Action Sanitaire et Sociale des Pyrenees Atlantiques in France was notified of an outbreak of acute respiratory illness among residents of a nursing home on 4 December, 1995. Over 50 of the 69 r PMID- 12631840 TI - EWGLI: a European surveillance scheme for travel associated legionnaire s disease. AB - The European Working Group for Legionella Infections (EWGLI) was set up in 1986 and introduced the European Surveillance Scheme for Travel Associated Legionnaires Disease in 1987. The microbiologists working in reference laborat ories and the epidemiolog PMID- 12631841 TI - Editorial. AB - Eurosurveillance this month presents European liaisons for the surveillance and control of communicable diseases at different stages of development. HELICS - the scheme for standardising the data collected about hospital acquired infections - has come and PMID- 12631843 TI - HELICS: a European project to standardise the surveillance of hospital acquired infection, 1994-1995. AB - During the 1990s several European countries began to set up national or regional networks for the surveillance of hospital acquired infections. Most of these networks were based on the US Centers for Disease Control and Prevention (CDC) National Nosocomia PMID- 12631842 TI - Mumps epidemic in Portugal despite high vaccine coverage - preliminary report. AB - A measles, mumps, and rubella (MMR) trivalent vaccine was added to Portugal's National Immunisation Programme (NIP) in 1987. All vaccines are given at health centres, free of charge, but an epidemic of mumps began in 1995, firstly in northern Portugal and PMID- 12631844 TI - Viral meningitis in Cyprus and England : summer 1996. AB - An outbreak of viral meningitis began in Cyprus on 5 July 1996. By 28 August a total of 316 cases had been admitted to hospital, most of whom were infants and young children; 55 (17%) were less than 1 year of age, 117 (37%) were aged 1 to 4 years, 103 (33 PMID- 12631846 TI - Eurosurveillance on the web. AB - Eurosurveillance is available electronically for readers with access to the internet on the world wide web at the home page of the European Centre for the Epidemiological Monitoring of AIDS. At the site a menu describes the bulletin and lists the particip PMID- 12631845 TI - The European Programme for Intervention Epidemiology Training. AB - The European Programme for Intervention Epidemiology Training (EPIET) provides practical experience in infectious disease epidemiology. EPIET aims to create a network of professionals throughout Europe trained to use a standard approach in intervention ep PMID- 12631847 TI - SRSV-1 gastroenteritis in Malta - 1995. AB - Small round structured viruses (SRSV) are known to cause epidemic gastroenteritis in institutions and account for 54% of person to person outbreaks and 6% of foodborne outbreaks in England and Wales. Infection is commonly transmitted from person to person PMID- 12631848 TI - The tuberculosis situation in Portugal : a historical perspective to 1994. AB - Portugal built its first tuberculosis (TB) hospital in Funchal, Madeira, in 1862. In 1899, Queen Amelia proposed and sponsored a private foundation, with clinics, hospitals, sanatoriums and preventoria, where relatives and other contacts of TB patients we PMID- 12631850 TI - European Concerted Action on Lyme Borreliosis (EUCALB). AB - Continuing uncertainty about the extent of the problem posed by Lyme borreliosis, the risk of serious sequelae, and unsatisfactory laboratory techniques to investigate it have led to the collaboration of researchers from 12 European Union (EU) countries a PMID- 12631851 TI - Airport malaria : a rare disease still poorly understood. AB - The article by Daniel Lusina and colleagues in this issue of Eurosurveillance is a timely reminder of the dangers of airport malaria. Since 1977, when the disease was first documented, 75 cases of airport malaria have been observed in western Europe, 28 o PMID- 12631849 TI - Prevalence of HIV-2 infection in Europe. AB - The HIV/AIDS pandemic is primarily caused by HIV-1. Another type of virus, HIV-2, is found mainly in certain West African countries In Europe, most of the cases of HIV-2 infection described have been in people from Africa. The objective of this study was PMID- 12631852 TI - Airport malaria : four new cases in suburban Paris during summer 1999. AB - During the summer 1999, four clustered cases of airport malaria were observed in France. The cases analysis revealed that airport malaria, which is a rare disease whose diagnosis is difficult, can be observed outside occupations at risk, in people livin PMID- 12631853 TI - Two outbreaks of mumps in children vaccinated with the Rubini strain in Spain indicate low vaccine efficacy. AB - This paper reports a descriptive study of two outbreaks of mumps in Spanish towns in 1998 and an assessment of the efficacy of the vaccine used. It provides evidence of the low level of protection against the mumps virus that has been observed previously PMID- 12631854 TI - Endemic presence of Salmonella enterica serotype Cerro in southern Italy. AB - Molecular typing of salmonella strains isolated between 1997 and 1999 in southern Italy and carried out by the Southern Italy Centre for Enteric Pathogens, has shown a high frequency of Salmonella enterica serotype Cerro. This serotype is extremely rare i PMID- 12631855 TI - Surveillance of influenza in Europe from October 1999 to February 2000. AB - Surveillance of influenza through EISS (European Influenza Surveillance Scheme) during the 1999 to 2000 winter shows that influenza affected most of the 11 participating countries and was particularly active in December 1999 and January 2000. Influenza A( PMID- 12631856 TI - Surveillance of haemolytic uraemic syndrome in children under 15 years of age in France in 1998. AB - Data from a national network of paediatric nephrology departments in France suggest that the incidence of haemolytic uraemic syndrome (HUS) in 1998 was 0.7 cases per 100 000 children aged under 15 years and that cases occur sporadically. Six out of 85 cas PMID- 12631858 TI - The maturation of Eurosurveillance. AB - The recent union of the monthly bulletin Eurosurveillance and its all-electronic sister Eurosurveillance Weekly into a unique project marked the start of a new stage in their development. Eurosurveillance began publication in 1996 and Eurosurveillance Wee PMID- 12631857 TI - Italy s new vaccination schedule reduced coverage of poliomyelitis vaccine in Naples. AB - Italy s health ministry introduced changes to the immunisation schedules for children on 7 April 1999, including immunisation against poliomyelitis. The changes included the replacement of the first two childhood doses of oral poliomyelitis vaccine (OPV) PMID- 12631859 TI - An outbreak of Norwalk like virus gastroenteritis in a nursing home in Rotterdam. AB - Sixty-two per cent of elderly and disabled residents of a Dutch nursing home (74/120) and 32% of staff (33/102) became ill in an outbreak of Norwalk-like viral gastroenteritis. The outbreak spread from person to person was supported by temporal clustering PMID- 12631860 TI - Mumps outbreak confined to a religious community. AB - One hundred and forty-four cases of mumps were notified in a religious community in London between July 1998 and April 1999 and the infection was confirmed in 82% of those who underwent saliva testing (51/62). Half of the clinical and half of the confirme PMID- 12631861 TI - Improved surveillance of tuberculosis. AB - In 1996 a consensus statement on the surveillance of tuberculosis in Europe, recommended the collection of data on all cases of tuberculosis at the local level. The statement, backed by the International Union Against Tuberculosis and Lung Disease, the Wo PMID- 12631862 TI - Feasibility of surveillance resistance to antituberculosis drugs :Europe, 1997. AB - Between 28% and 100% of isolates from ten countries from the EuroTB network in 1997 were reported with data on antituberculosis drug susceptibility. Drug resistance was commoner among foreign-born patients and in general among patients who had been treate PMID- 12631863 TI - The Spanish multidrug resistant tuberculosis network. AB - The network to monitor the spread of multidrug resistance tuberculosis (MDR-TB) in Spain based on genomic typing and set up in January 1998 benefits from the participation of about 90% of the laboratories of the national health system. Of the 94 MDR-TB pa PMID- 12631865 TI - The socioeconomic burden of hospital acquired infection. AB - Adult inpatients in common specialties who developed hospital acquired infection (HAI) remained in hospital 2.5 times longer, incurred hospital costs almost three times higher, and incurred higher general practitioner, district nurse, and hospital costs a PMID- 12631864 TI - Surveillance of tuberculosis in Ireland. AB - The first disaggregate data on tuberculosis in Ireland were collected in 1998. A total of 424 cases were notified (rate 11.7/100 000 population), of which 241 cases were culture positive and 122 were smear positive pulmonary cases. Only 35 cases were fore PMID- 12631866 TI - Methicillin resistant Staphylococcus aureus: toward a coordinated response to a continuing challenge. AB - Methicillin resistant Staphylococcus aureus (MRSA) represents a major public health challenge in many health care institutions worldwide. It is a common cause of outbreaks of cross infection and has become endemic in many regions where it adds to the morb PMID- 12631867 TI - Dutch measures to control MRSA and the expanding European Union. AB - The Dutch national policy for the prevention of spread of methicillin resistant Staphylococcus aureus (MRSA) has shown that it is possible to suppress and prevent MRSA from becoming endemic in hospitals. Implementation of effective measures against MRSA w PMID- 12631868 TI - MRSA in nursing homes in the Netherlands 1989 to 1998: a developing reservoir? AB - The prevalence of methicillin resistant Staphylococcus aureus (MRSA) in Dutch nursing homes in 1998 was higher than that found in 1989 to 1997. The increased prevalence of MRSA could lead to colonisation outside these nursing homes. A study of the prevale PMID- 12631869 TI - Methicillin resistant Staphylococcus aureus in German hospitals develop narrower patterns of antimicrobial resistance. AB - The prevalence of methicillin resistant Staphylococcus aureus (MRSA) in German hospitals has continued to increase, but the range of anti-microbials to which they are resistant has decreased. The trend cannot be attributed to changes in antibiotic consump PMID- 12631870 TI - European Antimicrobial Resistance Surveillance System (EARSS): susceptibility testing of invasive Staphylococcus aureus. AB - Over 400 laboratories participate in EARSS (European Antimicrobial Resistance Surveillance System) and send data to the National Institute of Public Health and the Environment (RIVM) in The Netherlands. Data on about 14 000 isolates of Staphylococcus aure PMID- 12631871 TI - HIV reporting in western Europe : national systems and first European data. AB - AIDS case reporting has been an essential tool for monitoring HIV infection in western Europe. Recent trends in AIDS have been affected by improved antiretroviral treatments that delay HIV disease progression, however, and no longer serve as indicators of PMID- 12631872 TI - A national measles epidemic in Switzerland in 1997 : consequences for the elimination of measles by the year 2007. AB - Clinical cases of measles have been reported to the Swiss Sentinel Surveillance Network (Sentinella) since June 1986 and measles infections have been monitored by the national laboratory notification system since January 1988. These two surveillance syste PMID- 12631873 TI - Measles : sub optimal situation in France. AB - The French sentinel network created in 1984 consists of general practitioners from the whole mainland, recruited on a voluntary basis. The members of the network follow up the weekly epidemiological surveillance of 10 health indicators (influenza-like PMID- 12631874 TI - Recent trends in gonorrhoea - an emerging public health issue ? AB - Neisseria gonorrhoeae infection remains one of the commonest bacterial sexually transmitted infections (STIs) in many developed countries. Its disproportionate burden on deprived, inner city populations and association with infertility have made its contr PMID- 12631875 TI - The number of gonococcal infections identified by the RENAGO network is increasing. AB - Surveillance of sexually transmitted infections (STIs) in France is based mainly on laboratory reporting, as STIs are not among the diseases notified by physicians. The network RENAGO (Reseau National du Gonocoque) was set up in 1986 to monitor national t PMID- 12631876 TI - Pertussis immunisation in Europe - the situation in late 1999. AB - The history of pertussis in European countries has clearly shown that immunisation decreases the morbidity associated with the disease. Inactivated whole cell pertussis vaccines (wP) have been used in some national vaccination programmes for over 30 years PMID- 12631877 TI - Pertussis eradication: a challenge for public health. AB - Pertussis and pertussis vaccination, particularly the almost simultaneous conduct of different clinical trials in various parts of Europe, has generated much debate in the past ten years. Nonetheless many important public health aspects still need to be c PMID- 12631878 TI - Marked decline in pertussis followed reintroduction of pertussis vaccination in Sweden. AB - Immunisation against pertussis with an acellular pertussis vaccine for children at 3, 5, and 12 months was included in the Swedish vaccination programme in January 1996, 17 years after the withdrawal of whole cell vaccine in 1979. Within months coverage r PMID- 12631879 TI - Pertussis surveillance in Switzerland, 1992 to 1997 - a large epidemic in 1994. What next? AB - Cases of whooping cough have been recorded by the Swiss Sentinel Surveillance Network Sentinella (SSSN) since June 1991 to monitor pertussis trends in Switzerland and to assess the current vaccination strategy. The Sentinella network consists of a sampl PMID- 12631880 TI - The incidence of pertussis in the Netherlands has remained high since an outbreak occurred in 1996. AB - The incidence of pertussis throughout the Netherlands, determined by notifications, rose suddenly in 1996 to reach 27.2 cases per 100 000 population, compared with 2.3/100 000 on average from 1989 to 1995. Earlier increases in 1989 1990 and 1994 reflected PMID- 12631881 TI - Surveillance is action! AB - Surveillance must lead to action: such is its philosophy and its public health justification. This statement is perfectly illustrated by the surveillance of legionellosis. The disease is transmitted by contaminated aerosols from a waterborne germ reservoi PMID- 12631882 TI - The first 48 hours of investigation and intervention of an outbreak of legionellosis in the Netherlands. AB - The Westfriese Flora is a one week flower show held every year in Bovenkarspel, in the north west of the Netherlands, with agricultural and consumer demonstrations on the same grounds. This year s show, held from 19 to 28 February, attracted 80 000 visito PMID- 12631883 TI - Epidemic cluster of legionnaires disease in Paris, June 1998. AB - From 29 June to July 1998, four cases of legionnaires disease in British citizens were reported to the Reseau National de Sante Publique (RNSP) by the statutory notification system (declaration obligatoire (DO)) and by theEuropean Surveillance Scheme for PMID- 12631884 TI - Surveillance of legionellosis within a hospital in northern Italy: May 1998 to September 1999. AB - The incidence of legionella infection in Europe overall in 1997 was 3.9 cases per million population, with the lowest rates reported by Malta and Norway and the highest by Denmark. Ninety cases were reported in Italy (1.5 cases/million), 20% of which were PMID- 12631885 TI - Travel associated legionnaires disease in Europe: 1997 and 1998. AB - The European Surveillance Scheme for Travel Associated Legionnaires Disease was set up by the European Working Group on Legionella Infections (EWGLI) in 1987 to identify cases of legionella infection in returning travellers and to detect outbreaks and cl PMID- 12631886 TI - Reuse of single-use sterile medical devices in Danish hospitals after report discourages it. AB - The extent to which sterile medical devices marked for single-use are reused was surveyed recently in 100 hospitals in Denmark, Finland, Norway, and Sweden. The same method was used in a survey of Danish hospitals two years earlier. This has enabled us PMID- 12631887 TI - Investigation of an outbreak of amoebiasis in Georgia. AB - Entamoeba histolytica, a protozoan parasite, occurs worldwide and 12% of the world's population are estimated to be infected. The prevalence of infection varies between 1% in industrialised countries to between 50% and 80% in tropical countries, where tra PMID- 12631889 TI - From epidemiological surveillance to European integration. AB - Between March and June 1999, 442 000 Kosovar refugees arrived in Albania. The national surveillance system was unprepared for this and an emergency communicable disease surveillance system was set up to detect and control potential outbreaks among the ref PMID- 12631888 TI - Sporadic infection caused by enterohemorrhagic d Escherichia coli O157. AB - In 1997 the Austrian Federal Ministry of Health established a reference laboratory for enterohaemorrhagic Escherichia coli at the federal public health laboratory in Innsbruck. The reference laboratory investigates sources of infection causing food poison PMID- 12631890 TI - Surveillance of communicable diseases among the Kosovar refugees in Albania, April-June 1999. AB - Albania, a country with 3.5 million inhabitants, is facing an economic and social crisis. The average per capita income is less than US$1000 per year. Since 1995 unemployment has increased by 2.7%. Poverty and migration are major constraints of the societ PMID- 12631891 TI - A case of tuberculosis on a long distance flight: the difficulties of the investigation. AB - Transmission of Mycobacterium tuberculosis from passenger to passenger aboard a long airplane flight was reported in 1996. Other cases were previously published, in particular by the Centers for Disease Control which had then suggested criteria for epidem PMID- 12631892 TI - Outbreak of Salmonella enteritis bongori 48:z35:- in Sicily. AB - Salmonella bongori 48:z 35 :- was first isolated from a lizard in Chad in 1966 and was classified as a biochemically atypical strain of the subgenus I of Kauffmann. Successively, some additional strains with different antigenic formulas but similar bioche PMID- 12631893 TI - Pork related trichinellosis: emergence of a new mode of transmission? AB - In this issue, Eurosurveillance publishes a report about two outbreaks of trichinellosis - of eight and 44 cases, respectively - that occurred in the German state of Northrhine-Westfalia from October to December 1998. Well conducted epidemiological invest PMID- 12631895 TI - Surveillance of respiratory pathogens and influenza-like illnesses in general practices - The Netherlands, winter 1997-98. AB - The Netherlands Institute of Primary Health Care (NIVEL) has coordinated the activities of a sentinel surveillance network of 43 general practices since 1970. These practices care for 1% of the Dutch population, a sample representative of the national pop PMID- 12631894 TI - Two outbreaks of trichinellosis in the state of Northrhine-Westfalia, Germany, 1998. AB - Illness or death from trichinellosis is statutorily notifiable in Germany. Between nought and ten cases were reported each year from 1987 to 1997. From November 1998 to January 1999, however, 52 cases of trichinellosis were identified by the public health PMID- 12631896 TI - Malaria surveillance in Italy: 1997 analysis and 1998 provisional data. AB - Malaria is the commonest imported infectious disease in Italy. Malaria was endemic throughout much of the country until it was eradicated nearly 50 years ago. Since then, a malaria surveillance system has been set up to detect locally acquired cases that PMID- 12631897 TI - The diversity of food poisoning and sources of contamination. AB - The three entirely different forms of food poisoning described in this issue of Eurosurveillance illustrate the extraordinary propensity of the organisms to survive and cause illness. Clostridium botulinum is an organism which produces its toxin (a toxin PMID- 12631898 TI - Outbreak oof calicivirus gastroenteritis associated with eating frozen raspberries. AB - Small round structured viruses (SRSVs - for example, calici-, astro-, and entero viruses) are the commonest causes of outbreaks of non-bacterial gastroenteritis worldwide. Transmission of SRSVs by water and by various foods - including salads, bakery prod PMID- 12631899 TI - Surveillance of foodborne botulism in Poland: 1960-1998. AB - Human foodborne botulism, in contrast to the other two forms of botulism (wound and infant botulism), is an intoxication that results when preformed botulinum toxin is ingested. Sporadic cases and family and general outbreaks occur when food products are PMID- 12631900 TI - Outbreak of Salmonella gastroenteritis among attendees of a restaurant opening in Grece, June 1998. AB - Until recently the system for reporting infectious diseases in Greece was inadequate, but a new laboratory reporting system was introduced in 1998, in which collaborating laboratories throughout Greece report each week by e-mail or fax using standard form PMID- 12631901 TI - Surveillance of salmonella: achievements and future directions. AB - This special issue of Eurosurveillance focuses on progress of the Enter-net network in the surveillance and management of outbreaks of human salmonellosis in Europe. This network has established a frequently updated international database and collaboratio PMID- 12631902 TI - The Enter-net international surveillance network - how it works. AB - Enter-net is an international network for the surveillance of human gastrointestinal infections, which monitors salmonellosis and Vero cytotoxin producing Escherichia coli (VTEC) O157, including their antimicrobial resistance. When the network began it in PMID- 12631903 TI - Emergence of multidrug resistant Salmonella thyphimurium DT 104 in the Czech Republic. AB - The Czech Republic is a country in central Europe with a population of about 10 million inhabitants. Salmonellosis is the most commonly reported infectious intestinal disease. Until 1989 about 10 000 cases were reported each year, and similar numbers of S PMID- 12631904 TI - Salmonella enteritidis in Western Europe 1995-98 - a surveillance report from Enter-net. AB - A decline in the incidence of Salmonella enteritidis infections in western Europe was identified by the Salm-Net (forerunner of Enter-net) salmonella database between 1993 and 1995, when the numbers of S. enteritidis isolates fell from 37 647 to 31 482. T PMID- 12631905 TI - Management of the investigation by Enter-net of international foodborne outbreaks of gastrointestinal organisms. AB - The free circulation of people, goods, and foods within the European Union (EU) states has increased the opportunity for international outbreaks of foodborne infections despite prevention efforts to reduce microbiological contamination of foodstuffs. The PMID- 12631907 TI - Leptospirosis in Portugal: epidemiology from 1991 to 1997. AB - The term leptospirosis describes a group of diseases caused by members of the order Spirochaetales, mainly the species Leptospira interrogans. Infection can be asymptomatic but disease of differing severity and clinical features (often suggestive of menin PMID- 12631906 TI - European Antimicrobial Resistance Surveillance System (EARSS): objectives and organisation. AB - Effective European surveillance must have the agreement and active involvement of all participants, concluded a European Union (EU) conference on the need for surveillance of resistant microorganisms (the microbial threat), held in September 1998 in De PMID- 12631908 TI - An outbreak of Salmonella enteritidis food poisoning from a commercially produced cheese. AB - Nine suspected cases of food poisoning were reported from three hospitals to the epidemiology and prevention service (Servizio di Epidemiologia e Prevenzione - SEP) of the local health authority in Naples district (Azienda Sanitaria Locale, ASL NA 4) betw PMID- 12631909 TI - A summary of occupationally acquired HIV infections described in published reports to December 1997. AB - Since the first case of documented seroconversion after a specific occupational exposure to HIV was reported in 1984, national or regional systems for the surveillance of occupationally acquired HIV infection have been developed in many industrialised cou PMID- 12631910 TI - Surveillance of occupational exposure to bloodborne pathogens in health care workers: the Italian national programme. AB - Health care workers (HCWs) face a serious risk of acquiring bloodborne infections, in particular hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV), all of which are associated with significant morbidity and mortality PMID- 12631911 TI - Accident compensation and occupationally acquired HIV infections in German health care workers. AB - Two hundred and eighty-six occupational transmissions of HIV had been reported worldwide by the end of 1997. A third of these cases (95), usually associated with seroconversion after a specific occupational exposure, were classified as definite occupation PMID- 12631912 TI - Widening horizons, opening doors. AB - This issue of Eurosurveillance includes a paper whose publication in the bulletin requires some explanation. Asthma is not an infectious disease as such, but acute viral infections are known to precipitate asthma attacks and people with asthma are more li PMID- 12631913 TI - Vaccination campaign for meningococcal disease in a rural area in the Netherlands - January 1998. AB - Neisseria meningitidis group C regularly causes epidemics in schools, universities, and army units, and in the past decade community outbreaks have been reported. Since a safe and effective polysaccharide vaccine is available the issue arises repeatedly PMID- 12631914 TI - Asthma trends in Switzerland: the Swiss Sentinel Surveillance Network, 1988-1996. AB - The setting up of the Swiss Sentinel Surveillance Network (SSSN) in 1986 has provided an excellent opportunity to study the distribution and determinants of asthma. The system provides morbidity data, which are representative of family practices in primar PMID- 12631915 TI - An outbreak of Salmonella enteritidis infection associated with iced cake. AB - Several people developed severe symptoms of gastroenteritis after attending a first holy communion banquet in Benevento, a town of 60 000 inhabitants in southern Italy. About 60 people had attended the banquet, held on 14 June 1998, between 1400 and 1800 PMID- 12631916 TI - Rare diseases deserve attention. AB - In August, Eurosurveillance published reports of two outbreaks of trichinellosis detected in 1998 in France and Italy and linked with horse meat imported from Yugoslavia. This issue, otherwise dedicated to botulism, includes a report of a third one. Repor PMID- 12631917 TI - Botulism in the European Union. AB - Botulinum toxin was first described as a potent neurotoxin in the late eighteenth century. Currently three main distinct clinical and epidemiological botulism syndromes are described - foodborne botulism, which results from the ingestion of food contamina PMID- 12631919 TI - Botulism in the United Kingdom. AB - Clostridium botulinum is a spore forming bacterium that grows in the absence of oxygen and is responsible for three main epidemiological categories of disease: foodborne, infant, and wound botulism. Foodborne botulism is an intoxication caused by ingestio PMID- 12631918 TI - Classic and emergent forms of botulism: the current status in Italy. AB - Botulism is a rare, severe, neuroparalytic disease. Four forms of botulism are described in humans: foodborne botulism and the more recently described wound botulism, infant botulism, and infant-like botulism. The two last forms are sometimes grouped un PMID- 12631920 TI - Botulism in Norway. AB - Botulism is a severe neuroparalytic disease caused by toxin produced by Clostridium botulinum, an anaerobic spore-forming bacillus. Physicians in Norway are required to notify the National Institute of Public Health (NIPH) of cases of botulism immediately PMID- 12631921 TI - New outbreak of trichinellosis in the Midi-Pyrenees region of France, September October 1998. AB - Le 6 octobre 1998 onze cas de trichinellose survenus, en France, dans les departements de la Haute Garonne et du Tarn ont ete declares aux autorites sanitaires de la region Midi-Pyrenees. Les toutes premieres informations fournies par les medecins suggera PMID- 12631922 TI - ESEN: a comparison of vaccination programmes - Part three : measles mumps and rubella. AB - Cet article est le dernier d une serie de trois articles portant sur la comparaison, dans huit pays (Danemark, Angleterre et Pays de Galles, Finlande, France, Allemagne, Italie, Suede et Pays-Bas), des programmes vaccinaux et de l impact epidemiologique d PMID- 12631923 TI - Outbreak of mumps associated with poor vaccine efficacy - Oporto Portugal 1996. AB - Vaccination against mumps in Portugal began in 1987, with the introduction of the combined measles, mumps, and rubella vaccine (MMR) in the national vaccination programme (Programa Nacional de Vacinacao: PNV) for both sexes at 15 months. In November 1990, PMID- 12631924 TI - Descriptive epidemiology of exanthems in the Rotterdam region January 1997 to June 1998. AB - The European Advisory Group on Immunisation has recommended that measles should be eliminated from Europe by the year 2007, a target accepted by National Immunisation Programme Managers for the World Health Organization (WHO) European Region countries. I PMID- 12631925 TI - Surveillance of tuberculosis in the WHO European Region 1995-1996. AB - The EuroTB programme for the surveillance of tuberculosis in Europe was set up in 1996 to collect, analyse, and disseminate data on tuberculosis cases notified in the World Health Organization (WHO) European Region. Following a feasibility study performed PMID- 12631926 TI - ESEN : a comparison of vaccination programmes Part two : pertussis. AB - This article is the second of a series of three, which compare vaccination programmes, immunisation schedules, vaccine coverage, and the epidemiological impact of vaccination for diphtheria, pertussis, measles, mumps, and rubella in eight countries (Denma PMID- 12631927 TI - Upsurge of hepatitis A in the Netherlands - early 1998. AB - Far more cases of hepatitis A were identified in the Netherlands in the first five months of 1998 than in the same period of 1997. In order to investigate the reasons for this upsurge we compared the notified cases of hepatitis A in the first five months PMID- 12631928 TI - Notified cases of legionnaires disease in France in 2001. AB - In 2001, 807 cases of Legionnaires' disease were reported to the Institut de veille sanitaire (French national public health centre). The incidence of the disease was 1.35 cases per 100,000 inhabitants, compared to a mean European incidence of 0,6 per 100,000. The median age was 59 years [16-97], the group aged more than 80 being the most affected. The sex ratio M/W was 3.1. The outcome of the disease was known in 69% of all cases, the case fatality ratio rating 19.9%. Among the contributing factors found in 558 cases, 11% had a cancer or blood disease, 12% received an immunosuppressant treatment, 10% were diabetic and 40% were smokers. In 2001, 13% (105/807) cases stayed in a hospital or a clinic during the incubation period, compared to 20% in 2000, and 11% were travel associated. PMID- 12631929 TI - The ICU-HELICS programme: towards European surveillance of hospital-acquired infections in intensive care units. AB - The creation of a database intended for the comparative analysis of the rates of hospital-acquired infections in the 15 countries of the European Union is among the objectives of the HELICS network (Hospitals in Europe Link for Infection Control through Surveillance). PMID- 12631930 TI - Epidemiology of tetanus in Italy in years 1971-2000. AB - The incidence of reported tetanus in Italy decreased from 0.5/100,000 in the 1970s to 0.2/100,000 in the 1990 s. During this period of time, the case-fatality ratio decreased from 68% to 39%. Italy has the highest reported number of tetanus cases in European countries. Elderly women are the most affected: the proportion of women aged over 64 years among cases has increased from 60% in the 1970s to 76% in the 1990s. Vaccination campaigns need to be conducted to target this group, and the surveillance of tetanus has to be improved to identify additional groups of population at risk. PMID- 12631932 TI - Profile and evolution of the chemosusceptibility of falciparum malaria imported into France in 2000. AB - In 2000, the chemosusceptibility of imported malaria was stable in France. All countries of infection considered, the bi-resistance to chloroquine and cycloguanil has not changed from 1996 to 2000. The monotherapy using quinine or mefloquine remains the first-line treatment to falciparum malaria. Resistance to these two antimalarials is rare in Africa and has not evolved over the past 15 years. PMID- 12631933 TI - Spread of an MRSA clone with heteroresistance to oxacillin in the Netherlands. AB - Since 2001, an increase in the number of MRSA strains with heteroresistance to oxacillin has been observed in the Netherlands. These strains can spread unnoticed, as their phenotypic heterogenous resistance to beta-lactams affects the results of susceptibility testing. Therefore, further testing using PBP2a latex agglutination or PCR are recommended. PMID- 12631934 TI - Vaccination campaign following an increase in incidence of serogroup C meningococcal diseases in the department of Puy-de-Dome (France). AB - In the department of Puy-de-Dome, France, 17 cases of invasive meningococcal disease C were notified between March 2001 and the first week of 2002. Among the 15 confirmed cases, 11 (73%) were serogroup C, 2 (13%) serogroup B, and 2 could not be identified. The rapid increase in the number of cases in a period of low endemicity for the rest of the country and the severity of the disease (case fatality ratio 27%, purpura fulminans 64%) led the health authorities to initiate a vaccination campaign targeting children and young adults from 2 months up to 20 years living in a limited area of the department. Around 80,000 people were immunised between 16/01/02 and 09/02/02. More than half of the 1390 immediate side effects were headache and dizziness. As of mid-March, no further case of meningococcal disease has been notified since 6 January. PMID- 12631935 TI - The WHO European Region declared free of polio. AB - The WHO European Region* has been certified "free of poliomyelitis" at a meeting held in Copenhaguen, Denmark, on 21 June 2002. The information was released by the European Regional Commission for the Certification of Poliomyelitis Eradication. PMID- 12631936 TI - A national training programme in field epidemiology launched in France. PMID- 12631937 TI - Roles and functions of a European Union Public Health Centre for Communicable Diseases and other threats to health. AB - An international consensus has been reached that a European Union (EU) Technical Coordination Structure (TCS) for communicable diseases is needed to improve Europe s future response to international communicable disease threats within and beyond its boundaries. After the American events of September 11 2001 and the deliberate releases of anthrax, the EU created a Health Security Committee, adopted a civil protection decision, and established for 18 months a team to develop responses for deliberate releases of biological and chemical agents. These two initiatives, the network s approach and health security work, must converge into a single stream addressing health protection for the people of Europe. They could be combined into a European Centre for Communicable Diseases that is planned to become active by 2005. PMID- 12631938 TI - Measles in France: the epidemiological impact of suboptimal immunisation coverage. AB - The promotion of immunisation in France since 1983 has resulted in a 97% reduction in morbidity and a reduction of 60% of mortality. However, the stable and sub-optimal coverage around 84% leads to a shift in higher age groups, where complications are more frequent and serious. The proportion of those aged over 10 years was 13% in 1985 and reached 48% in 1997, the transmission of measles being maintained in France. To eliminate the disease, vaccine coverage with 2 doses and over 95% would be necessary. PMID- 12631939 TI - Laboratory capability in Europe for foodborne viruses. AB - This report describes a survey of national laboratory capabilities of diagnostics and surveillance databases for foodborne viruses among the "Foodborne Viruses in Europe" consortium. All the countries have laboratories that can test for HAV antibody in human serum. Eight of the ten surveyed European countries maintain a national database of HAV cases. Food can be tested for the presence of HAV in Finland, Italy, Spain, France and Denmark. All surveyed countries have at least one laboratory that tests for Norwalk-like virus (NLV) by reverse transcriptase polymerase chain reaction and all also have the capability to use electron microscopy. Five countries maintain a national database of NLV cases and nine maintain a national database of NLV outbreaks. Almost all participant countries have laboratories that can test for NLV in food items including shellfish. PMID- 12631940 TI - Occurrence of Salmonella enterica serovar Dublin in Austria. AB - In Austria, Salmonella enterica subsp. enterica serovar Dublin, a bovine-adapted serovar, rarely causes infections in humans. In 2000, Austria was within the European mean with an incidence of 0.1 per million inhabitants. Our data show that the vast majority of all serovar Dublin infections (human and non-human) can be traced epidemiologically to two districts in the Tyrol. This concentration of cases can be explained by a particularly traditional aspect of cattle farming in this area, the alpine pasture. There is an increased risk of cross infection due to the communal keeping of animals from various farms. Infected cattle are a source of infection for humans, and transmission usually occurs from eating beef and drinking cows milk. Using pulsed field gel electrophoresis and automated ribotyping, three out of five isolates from human infections could be traced to characteristic Tyrolean Dublin clones. Bacteriological screening for faecal carriage before the transfer of cattle from risk-herds to the alpine pastures and before the return from risk-pastures to the farms would be a possible starting point to prevent cross-contamination of large mixed herds and contamination of pasture through latently infected cattle. Appropriate research is necessary. PMID- 12631941 TI - Viral haemorrhagic fevers in Europe--effective control requires a co-ordinated response. AB - Viral haemorrhagic fevers (VHF) have attracted the attention of the medical world and general public for many reasons, some based in reality and more on misinformation. They are amongst the highest profile infections in the public mind, because they are thought to be highly infectious and to kill most of their victims in a dramatic way (1,2). To add to the intrigue, mysteries remain about the source of some of the viruses involved. They emerge and re-emerge in many countries, most recently Ebola in Uganda in 2000 (3) and Gabon in 2001/02 (4), and Congo Crimean Haemorrhagic Fever (CCHF) in Kosovo (5) and Pakistan in 2001 (6). Large outbreaks have affected populations in endemic areas, living mainly in inaccessible areas or refugee camps where living conditions are very difficult. Poorly resourced medical facilities have played a role in amplifying transmission and infection control measures have been difficult or virtually impossible to establish. These viruses are likely to remain a threat until the reservoir is identified and as long as endemic areas are afflicted with ecological change, poverty and social instability. Recent events since September 11 2001 remind us of their potential to be used as weapons, and that fear can present a risk to public health. PMID- 12631942 TI - Ebola in Africa--discoveries in the past decade. AB - Within the past decade, Ebola haemorrhagic fever (EHF) has been recognised for the first time in four countries. Our understanding of the epidemiology, clinical aspects, laboratory diagnosis and control measures for EHF has improved considerably as a result of the outbreaks in these countries and the re-emergence that has occurred in another. The coordinated international responses to several of the large EHF outbreaks serve as models for controlling epidemics of other communicable diseases. This report is a chronological overview of the EHF outbreaks in Africa during the past decade, including the recent epidemics in Gabon and the Republic of the Congo, and highlights new discoveries and some of the remaining challenges. PMID- 12631943 TI - Management of patients in Germany with suspected viral haemorrhagic fever and other potentially lethal contagious infections. AB - Patients suffering from viral haemorrhagic fevers must be handled specifically. The clinical diagnosis of these diseases in the initial stage is difficult because early symptoms are non specific. In Germany, specific diagnosis is available at two diagnostic centres with biosafety level 4 facilities. Five high security infectious disease isolation units for patient care are available in Munich, Leipzig, Hamburg, Berlin, and Frankfurt. In addition, a corresponding number of centres of competence are established to offer support and advice to the hospitals initially treating the patients and to the local public health officers. The decentralisation of these centres of competence is recommended to allow for more timely and reactive responses to VHF epidemic threats. The risk categorisation for contacts has proved to be very useful in practice. PMID- 12631944 TI - Management of viral haemorrhagic fevers in Switzerland. AB - Over the past years, there have been very few imported cases of VHF in Switzerland: one confirmed and four suspected cases of Ebola fever in Basel in 1994, two suspected cases of Ebola and Lassa fevers in Lausanne in 2000, and in the same year, six suspected cases of Lassa fever in Geneva. Given the considerable diversity in the management of patients with suspected or confirmed VHF, national guidelines are needed, as well as the establishment of a national reference centre. PMID- 12631945 TI - Current management of patients with viral haemorrhagic fevers in the United Kingdom. AB - In the UK, suspected and confirmed cases of viral haemorrhagic fever are currently managed according to the 1996 Guidance of the Advisory Committee on Dangerous Pathogens, which describes an approach to the risk categorisation of suspected cases. It also provides guidance on patient management including transfer, laboratory investigations, infection control, and monitoring of contacts based on the risk assessment. Confirmed cases are managed in bed isolators ("Trexler units"), two of which are available in high security infectious disease units in the UK. This guidance is under review and may change. Recent experience has shown that communication and reassurance for health care workers and the public are major tasks in managing such cases. PMID- 12631946 TI - Management of viral haemorrhagic fever in the Netherlands. AB - Two cases of Lassa fever have been reported in the Netherlands since viral haemorrhagic fevers became notifiable diseases in 1978. In 1980, an expatriate from Burkina Faso who was not seriously ill was confirmed by laboratory tests after his discharge from hospital. The second case occurred in 2000: the patient died on the 11th day of admission to hospital. The problems we faced in the management of this case and the contact investigation--more than one hundred contacts - highlighted the need for national recommendations in the Netherlands. PMID- 12631947 TI - Management of Lassa fever in European countries. AB - The UK Public Health Laboratory Service hosted a meeting in London during November 2000 to discuss the management of Lassa fever following the four cases which had occurred in 1999/2000. Participants were invited because they had recent clinical or laboratory experience of managing cases of Lassa fever or had been involved in public health aspects of a case of this infection. The meeting was closed to enable free and frank discussion of confidential matters. This is a report of the general issues and recommendations which arose from the discussion. PMID- 12631948 TI - A new decline in preventive behaviours among homosexual men : the role of highly active antiretroviral therapy? AB - The articles published in this issue of Eurosurveillance on recent trends of preventive behaviours in homosexual men in Spain, Switzerland, and the Netherlands, all conclude that the practice of safer sex is declining. They emphasise increases in anal penetration (regardless of the nature of the relationship), non-protected anal intercourse, and ejaculation in the mouth, in particular among casual partners. The increase of diagnoses of acute sexually transmitted diseases (such as gonorrhoea and syphilis) in different European countries - illustrated in this issue by the data from the Netherlands - is the first tangible consequence of this decline. PMID- 12631949 TI - Men having sex with men and HIV/AIDS prevention in Switzerland: 1987-2000. AB - Although the latest survey on prevention behaviours of men having sex with men (MSMs) in Switzerland shows a decreasing number of HIV cases since 1992, a decrease in the protection level has also been observed. This could lead to a higher rate of HIV notifications in 2001 compared to 2000. PMID- 12631950 TI - A summary report from Amsterdam: increase in sexually transmitted diseases and risky sexual behaviour among homosexual men in relation to the introduction of new anti-HIV drugs. AB - The worrying increase of sexually transmitted diseases (STDs) in Amsterdam was investigated in two studies carried out by the department of AIDS research of the municipal health service. The results indicate that the introduction of Highly Active Antiretroviral Therapies (HAART) may have had an influence on the increase of STDs and risky sexual behaviours in Amsterdam. PMID- 12631951 TI - Monitoring HIV prevalence and behaviour of men who have sex with men in Barcelona, Spain. AB - Recently, different studies among men who have sex with men (MSMs) have reported an increase in HIV incidence and sexually transmitted infections, and an increase in sexual risk behaviour. But the optimism regarding anti-retroviral treatments may lead to a greater relaxation in protective measures in the near future. PMID- 12631952 TI - Influenza pandemic: preparedness planning in Germany. AB - The following conceptual framework formed the basis for a common decision made by the health ministers of Germany's 16 federal states to set up an influenza pandemic preparedness plan. The worst case scenario was used, on the basis of the data from the pandemic of 'Spanish flu', in 1918-20. The priority groups for vaccination were assessed, as well as the potentially available antiviral treatments. National policies could be highly improved by a common European view. PMID- 12631953 TI - Trend of multiple drug resistant Salmonella Typhimurium in Norway. AB - This article reports the trend of multidrug resistant Salmonella Typhimurium isolated from humans in Norway from 1998 to 2000. Most of the incidents with multiple resistant S. Typhimurium infection contracted in Norway have been DT104. We should therefore expect an increase of both primary and secondary infections if strains such as phage type DT104 with R-type ACSSuT become endemic. PMID- 12631954 TI - Report on the Sixth International Meeting of the European Laboratory Working Group on Diphtheria, Brussels, Belgium. AB - In addition to the Eastern European resurgence of diphtheria during the last decade, there has also been an emergence of infections caused by non-toxigenic Corynebacterium diphtheriae and non-toxigenic, toxin gene bearing C. diphtheriae. Given that these strains may manifest as symptomatic infections of differing degrees of severity, their clinical and epidemiological significance need to be assessed. The persistence of toxigenic and non-toxigenic C. diphtheriae in circulation, together with genotypic and biotype variability means that innovative measures to vaccinate populations are pertinent. The most effective method of protecting the currently most vulnerable population group (adults) is to implement a booster dose of vaccine amongst the adult populations. Furthermore, in combination with an efficient surveillance system, effective antibiotic prophylaxis and an up-to-date vaccination programme, serological studies needs to be maintained to monitor the immunity status of the population. PMID- 12631955 TI - Confirmed interruption of indigenous measles transmission in Catalonia. AB - From November 1998 to December 2000, 84 suspected measles cases were reported in Catalonia (6 090 040 inhabitants). Of the 73 laboratory tested cases (87%), 20 showed IgM antibodies specific to measles and 3 were epidemiologically linked to a confirmed case. Among these 23 confirmed cases, 13 were indigenous, the last two cases reported dating back from June 1999 and July 2000. These results confirm the success of the measles elimination programme implemented in 1998 in Catalonia. PMID- 12631956 TI - Evolution of antibiotic resistance of non-typhoidal salmonellae in Greece during 1990-97. AB - Susceptibility to 15 antibiotics was determined in 1548 non-typhoidal salmonella strains isolated in Greece from l990 to l997. The overall prevalence of resistance of both Salmonella enterica serotype Enteritidis and Salmonella enterica serotype Typhimurium increased during the first years of the study. A decrease was observed from 1996, especially for S. Enteritidis, which showed the highest overall antibiotic resistance. S. Typhimurium was the serotype with the highest multiresistance to antibiotics. The rest of the serotypes had very low resistance prevalence compared with both S. Enteritidis and Typhimurium serotypes. PMID- 12631957 TI - An outbreak of legionnaires disease in a municipality in northern Portugal. AB - Eleven cases of legionnaires disease, all patients living in the same municipality, were admitted to a district hospital in northern Portugal. Preliminary investigations suggested an association with the events of the annual festivities in that municipality. Legionella pneumophila was not isolated from any of the suspected sources, but evidence from a case control study suggested that an aerosol produced by a decorative fountain in the main square during the night of a rock concert was the likely vehicle of infection. The prevalence of smoking was higher among cases than controls. PMID- 12631958 TI - Surveillance of antimicrobial resistance in Denmark. AB - Recent data of the Danish Integrated Antimicrobial Resistance Monitoring and Research Programme (DANMAP) show that, in Denmark, resistance levels among Salmonella enterica are modest and that resistance in Escherichia coli isolates causing disease in anim PMID- 12631959 TI - FiRe works - the Finnish Study Group for Antimicrobial Resistance (FiRe). AB - The Finnish Study Group for Antimicrobial Resistance (FiRe), a network founded in 1991, has established a standard routine susceptibility testing method that yields reliable and comparable results for both diagnostic and epidemiological purposes. Results PMID- 12631960 TI - Bacterial resistance to antibiotics in France : a public health priority. AB - For a few years, France has been faced to a rapid spread of anti-microbial resistance in hospitals and in general practice despite the many recommendations issued to solve this problem. In 1999, the Institut de Veille Sanitaire conducted a collective expe PMID- 12631961 TI - An outbreak of legionnaire s disease among visitors to a fair in Belgium, 1999. AB - Ninety-three cases of legionnaires disease (43 confirmed, 12 presumptive, and 38 possible/ clinical) were identified in an outbreak associated with a trade fair in Kapellen, Belgium in November 1999. Five cases died. Epidemiological investigation showed PMID- 12631962 TI - Development of a research netwrok for emerging foodborne pathogens in Germany. AB - A network of epidemiologists, microbiologists, veterinarians, and clinicians is being developed in Germany to support local and state health departments and veterinary authorities in their research and epidemiological investigations on emerging foodborne PMID- 12631963 TI - Salmonella enterica serotype Oranienburg infections associated with consumption of locally produced Tyrolean cheese. AB - Sixteen culture confirmed cases of enteric infection with Salmonella enterica serotype Oranienburg were detected between August 10 and September 29 1999 in Tyrol (Austria). Ten of them suffered bloody diarrhoea and six were asymptomatic carriers. Intervie PMID- 12631964 TI - Towards optimization of surveillance resistance to antituberculosis drug resistance. AB - In the past decade, strengthening tuberculosis (TB) surveillance has become a public health priority in Europe. In western Europe, TB case rates slowed their previous decline, but in eastern Europe dramatic increases, often with doubling of rates, were re PMID- 12631965 TI - European recommendations on surveillance of antituberculosis drug resistance. AB - Antituberculosis drug resistance, whose extent in Europe is not well documented, is a serious threat to tuberculosis control. The aim of the recent European recommendations on antituberculosis drug resistance surveillance, issued by a working group compos PMID- 12631966 TI - BSE : the European regulatory context. AB - The Bovine Spongiform Encephalopathy crisis provoked a fundamental re-appraisal of the way in which the European Community approaches matters of food safety. Between 28 July 1989, when restrictions on the dispatch of certain live cattle from the UK starte PMID- 12631968 TI - Surveillance of Creutzfeldt Jakob disease in the United Kingdom. AB - By the end of May 2000, 54 definite cases and 13 probable cases of vCJD had been notified in the United Kingdom by the National CJD Surveillance Unit set up in 1990. All definite cases for whom data are available are methionine homozygous at codon 129 of PMID- 12631967 TI - Developments in variant Creutzfeldt Jakob disease. AB - The announcement on 17 July 2000 that the rate of increase in the incidence of variant Creutzfeldt Jakob disease (vCJD) in the United Kingdom (UK) had reached statistical significance makes the current issue of Eurosurveillance particularly timely. A clus PMID- 12631969 TI - New variant of Creutzfeldt Jakob disease. AB - Work on experimental pathology carried out for over 10 years indicates that the biological properties of the BSE agent, responsible for the new variant of Creutzfeldt-Jakob disease (vCJD), are particular. In contrast to the situation observed in the other PMID- 12631970 TI - Epidemiological status of BSE in France - update on born after the ban cases. AB - Despite measures taken in France since 1990 to control the spread of bovine spongiform encephalopathy (BSE), the incidence of BSE cases notified in France by the BSE Epidemiological Surveillance Network has been rising in recent years. By 1 July 2000, 103 PMID- 12631971 TI - Salmonella - "old" organism, continued challenges ! PMID- 12631972 TI - Explosive increase of Salmonella Java in poultry in the Netherlands: consequences for public health. AB - In the Netherlands Salmonella Paratyphi B variant Java increased in poultry from less than 2% of all isolates before 1996 to 60% in 2002. Despite exposure to contaminated meat is high, human patients with Java infection are rare (0.3% of all isolates). However, 50% of the human isolates showed PFGE profiles identical to the poultry clone. Resistance to flumequin in S. Java increased from 3% between 1996-2000 to 19% in 2001, and 39% in 2002, while that of other serotypes in poultry remained at about 7%. S. Java is also fast becoming less sensitive to ciprofloxacin. PMID- 12631973 TI - Investigation of human infections with Salmonella enterica serovar Java in Scotland and possible association with imported poultry. AB - PFGE analysis of S. Java strains (29 from humans, 30 from poultry meat) showed two major clusters. All isolates from poultry imported from the Netherlands belonged to Cluster A, which also comprised 10 human isolates. Thirty-one of the 37 isolates in this cluster had an identical JavX1 pattern, similar to the X8 profile of a particular S. Java clone predominant in poultry production in several European countries. Cluster B comprised 19 human isolates and two poultry isolates of unknown origin. These results combined with epidemiological data and information on the origins of poultry meat strongly suggested that imported poultry meat is an important source of Java infections in humans in Scotland. PMID- 12631975 TI - The Salm-gene project - a European collaboration for DNA fingerprinting for food related salmonellosis. AB - An external quality assessment of PFGE method to discriminate between salmonella serotypes and lysotypes was carried out by the Salm-Gene project in Europe. A set of 16 strains of S. Enteritidis was sent to 9 national salmonella reference laboratories. By using a harmonised protocol, the PFGE profiles produced were comparable between each centre. In most cases, there was at least 90% similarity between isolates tested in the different European laboratories and there was usually >95% similarity. This suggests that PFGE analyses are reproducible and therefore can be used as a valuable investigation tool combined with epidemiological data. PMID- 12631974 TI - Antimicrobial drug resistance in isolates of Salmonella enterica from cases of salmonellosis in humans in Europe in 2000: results of international multi-centre surveillance. AB - The Enter-net surveillance system received results of antimicrobial sensitivity tests for isolates from over 27 000 cases of human salmonellosis in 2000 in 10 European countries. Almost 40% of isolates were resistant to at least one antimicrobial, with 18% multiresistant. Resistance to ampicillin, streptomycin, sulphonamides and tetracyclines was common, with over 20% of isolates resistant to at least one of these antimicrobials. Clinical resistance to ciprofloxacin was rare, with only 0.5% of isolates exhibiting such resistance (MIC >1.0 mg/l). Resistance to nalidixic acid coupled with a decreased susceptibility to ciprofloxacin (MIC 0.25-1.0 mg/l) was more common, with 14% of isolates showing these properties. Resistance to third-generation cephalosporins was rare with only 0.6% of isolates resistant to cefotaxime. In all countries multiple resistance was most common in Salmonella enterica serotype Typhimurium, with 51% of isolates multiresistant in total. In England and Wales multiple resistance was also prevalent in S. Virchow and S. Hadar, whereas in other countries multiple resistance was common in serotypes such as S. Blockley. PMID- 12631976 TI - Salmonella enteric infections in Gipuzkoa, Spain, 1983-2000. AB - The incidence of Salmonella enteric infections in Gipuzkoa, Spain, was estimated by studying a stable population between 1983 and 2000. Only stool culture confirmed cases were included. The annual mean rate of infection in children under 2 years old was 1121 per 100,000 (CI 95%; 1060-1181). This age group had the highest relative risk (RR), 16.2-fold higher than the RR of those aged over 14 years. Salmonella Enteritidis was the most prevalent serovar (80.4% of all patients), followed by Salmonella Typhimurium (11.7%). PMID- 12631977 TI - An outbreak of Norwalk-like viral gastroenteritis in holidaymakers travelling to Andorra, January-February 2002. AB - A retrospective cohort study was conducted to investigate an outbreak of Norwalk like viral gastroenteritidis that occurred in Irish holidaymakers visiting Andorra, in January-February 2002. Preliminary results showed the risk exposure was higher for tourists who stayed in Soldeu and consumed ice cubes in their drinks (OR = 2.5, 95% CI [1.3-4.6)], after logistic regression and adjusting for sex and water consumption). PMID- 12631978 TI - Patterns of Puumala virus infection in Finland. AB - Puumala hantavirus infection is prevalent throughout most of Europe, and in endemic areas it may be the most common cause of acute renal failure. To evaluate trends in incidence of Puumala virus infections in Finland, we analysed national surveillance data in 12-month periods from March 1995 to February 2002. During this time, 8184 laboratory-confirmed cases were notified to the National Infectious Disease Register. Three epidemic periods were identified, for which the number of cases was more than 1400 (there were approximately 600-900 cases per non-epidemic period). The incidence of Puumala hantavirus infection varied by geographic region during the study period, and the overall number of cases may be increasing. PMID- 12631979 TI - Creutzfeldt-Jakob disease and related diseases in France from 1998 to 2000. AB - In France, the number of reports of suspected CJD increased from 1998 to 2000, probably due to the increase in the requests for biological tests: respectively 459 in 1998, 590 in 1999, and 823 in 2000. For all three years, the distribution by sex is similar, with a sex ratio (M/F) of 1.05. The proportion of suspected cases aged under 50 remained stable (16% of all reports in 1998-2000). The number of sporadic CJD, confirmed or probable, was stable, with a mortality ratio of 1.38 per one million in 1998, 1.56 in 1999, and 1.41 in 2000. PMID- 12631980 TI - Tuberculosis in Austria 1995-99: geographical distribution and trends. AB - A descriptive analysis of routine surveillance data on tuberculosis (TB) from 1995 to 1999 was performed in Austria. The federal states of Vienna and Upper Austria showed the highest yearly incidences for all five years. In general, however, a decrease of the yearly incidence was observed for all federal states except for Carinthia (where there was a small increase of 3.2 %). In the cities of Vienna, Linz, Wels, Salzburg, Klagenfurt and St-Polten, as well as in the regions of Eastern Tyrol, Southern Carinthia, and Southern Burgenland, the incidence was above average (>19 cases/100,000 for each). The number of cases per year and per district reported by the statutory reporting system were clearly in excess of those reported by hospitals. The comparison of the two datasets, however, showed a highly significant regional correlation of age standardised morbidity rates at district level. The findings are prone to reporting bias and might not reflect the real TB situation. The responsible health authorities should therefore consider the results as a 'working hypothesis in order to review the reporting behaviour at district level. PMID- 12631981 TI - Influenza under close surveillance. PMID- 12631982 TI - Outbreak of influenza, Madagascar, July-August 2002. AB - Preliminary investigation found that a large outbreak of influenza-like illness occurred in Madagascar during July-August 2002, with 30 304 cases and 754 deaths reported. Most cases were reported from the highland regions of Fianarantsoa Province, in centre Madagascar. The majority of the cases lived in rural areas, and children under five years and adults 60 years and older were the most affected. The outbreak was attributable to an A/Panama/2007/99-like (H3N2) virus, which has been circulating worldwide for several years. PMID- 12631983 TI - Surveillance of influenza in the North-West Region of England 2001-02. AB - A local sentinel network of general practitioners has been established in the north west of England for the surveillance of influenza. In the 2001-02 winter, consultation rates for influenza-like-illness (ILI) were low but the surveillance network was able to demonstrate sub-regional variations in the timing of peak influenza activity, and the infection of different age groups. This suggests the network can contribute to better planning to winter pressures on the North West health service. PMID- 12631984 TI - Enhancing the predictive value of throat swabs in virological influenza surveillance. AB - According to a national survey in Germany, the influenza virological surveillance can be improved when sentinel practitioners take throat swabs specimens only from patients who consult early after the disease onset (ie, within 48 hours), and when they use the strict clinical case definition of influenza-like illness. PCR should be used for primary detection of influenza viruses. PMID- 12631985 TI - Real-time modelling of influenza outbreaks--a linear regression analysis. AB - Seasonal outbreaks of influenza exert a considerable burden on health services, and are notorious for their variability from year to year. Making use of historical data from the Scottish sentinelle surveillance since 1972, a potential candidate model has been derived based on simple linear regression. It was applied with a measure of success in the 1999-2000 winter season. PMID- 12631986 TI - Spanish flu epidemic in 1918 in Geneva, Switzerland. AB - In Geneva, Switzerland, the Spanish flu epidemic affected more than 50% of the population. The mortality was higher among those aged between 20-49 years and among men. The socioeconomic impact was very important, as the outbreak led to severe dysfunctions, including in health services. This epidemic shows the socio economical burden that may be associated with influenza and highlights the need for pandemic preparedness. PMID- 12631987 TI - Mild to moderate influenza activity in Europe and the detection of novel A(H1N2) and B viruses during the winter of 2001-02. AB - Influenza activity in Europe during the 2001-02 influenza season was mild to moderate. Compared to historical data, the intensity was low in six countries, medium in eleven and high in one country (Spain). The dominant virus circulating in Europe was influenza A(H3N2). Two novel influenza virus strains were isolated during the 2001-02 season: influenza A(H1N2) viruses (mainly isolated in the United Kingdom and Ireland, but also in Belgium, France, Germany, the Netherlands, Portugal, Sweden, Switzerland and Romania), and influenza B viruses belonging to the B/Victoria/2/87 lineage (mainly isolated in Germany, but also sporadically in France, Italy, the Netherlands and Norway). With the exception of H1N2 virus detections in England, and Ireland and the influenza B viruses belonging to the B/Victoria/2/87 lineage in Germany, these two viruses did not circulate widely in Europe and did not play an important role in influenza activity during the 2001-02 season. An influenza B virus belonging to the B/Victoria/2/87 lineage will be included in the 2002-03 influenza vaccine. The new subtype influenza A(H1N2) is covered by the 2002-03 vaccine, as the haemagglutinin and neuraminidase components of the H1N2 viruses are antigenically similar to the vaccine components (H1N1 and H3N2). PMID- 12631988 TI - Development of a protocol to evaluate the quality of clinical influenza data collected by sentinel practitioners in Europe. AB - EISS implemented an evaluation protocol to assess the quality of clinical data collected by sentinel networks for the surveillance of influenza. The protocol was applied in Belgium and in three regions in Spain where it resulted in recommendations to improve the national and regional surveillance networks. PMID- 12631989 TI - Quality control assessment of influenza and RSV testing in Europe: 2000-01 season. AB - The Quality Control Assessment (QCA) was initiated to evaluate the quality of the influenza and respiratory syncytial virus (RSV) testing in the national reference centres belonging to the European Influenza Surveillance Scheme (EISS) network. Samples were coded and sent in two panels of 12 samples within a two week interval to 16 laboratories during the 2000-01 winter season. The antibodies titration by HI test was reported by 60% of the laboratories (n=16), and the results were correct for 56% of them. One false detection of influenza B antibodies was reported by one laboratory, and for the others the sensitivity varied widely. The sensitivity of the tests for the detection of influenza virus varied for A(H3N2) from 10 to 100,000 TCID50/ml. The influenza A subtyping was performed by 87% of the laboratories, and 31% gave correct results. The characterisation of the variants was undertaken by six laboratories and half of them fully achieved it. Fifty six percent of the laboratories used RT-PCR for the diagnosis; the results were specific and the sensitivity equivalent to the cell culture. PMID- 12631990 TI - Influenza pandemic preparedness and response planning at community level. PMID- 12631991 TI - Clustered cases of leptospirosis in Rochefort, France, June 2001. AB - Five clustered cases of leptospirosis were diagnosed in the area of Rochefort, France, in June 2001, among teenagers who had swum in the Genouille canal. The symptoms included fever, headache, abdominal pain and vomiting, chills and myalgia. Three cases were confirmed by PCR and serology. The mean cumulative duration of bathing was significantly higher in cases (23.8 hours) compared to controls (14.4 hours). No other particular risk factor was observed. The environmental investigation revealed the presence of rodents excreting of leptospires near the bathing area. For all antigens considered, the occurence of seropositive rodents was 30.8%, L. icterohaemorrhagiae being the predominant serogroup (23,1%). PMID- 12631992 TI - Outbreak of trichinellosis in Caceres, Spain, December 2001-February 2002. AB - An outbreak of trichinellosis by T. britovi occured in Caceres, Spain, between 18 December 2001 and 11 February 2002, following the consumption of insufficiently cooked meat from a domestic pig. Among the 56 people exposed, 26 cases of trichinellosis were diagnosed, of which 17 serologically confirmed. The mean incubation period was 23.5 days (3-45). Among the foodstuffs suspected, salami type sausages were associated with an attack rate of 93.3% (14/15), and a dose response relationship was observed. Trichinella britovi is essentially a sylvatic species, but this study suggests a change in the epidemiology of trichinellosis. PMID- 12631993 TI - An outbreak of diarrhoeic shellfish poisoning in Antwerp, Belgium. AB - In Antwerp, Belgium, 403 cases of diarrhoeic shellfish poisoning were reported after consumption of blue mussels. Symptoms included diarrhoea, vomiting, abdominal pain, and nausea. The analysis of faecal specimens from patients allowed diagnosis exclusions for bacteria and viruses. Mouse-assays revealed the presence of biotoxins specific of dinoflagellates, which were identified and quantified by LC-MS. The mussels were imported from Denmark, and were part of a batch presenting high concentrations of okadaic acid above the regulatory limits. PMID- 12631994 TI - The Danish Statens Serum Institute celebrates its centenary. AB - The Statens Serum Institute (SSI) in Denmark celebrated its centenary on 9 September 2002. Conferences were organised on emerging and re-emerging diseases, vaccine development, and the launch of a new website (www.ssi.dk). PMID- 12631995 TI - The WHO European report confirms links between socioeconomic development and health. AB - Over 300 representatives, including 20 ministers of health, attended the 52nd session of the World Health Organisation (WHO) Regional Committee for Europe, which was held from 16 to 19 September in Copenhagen, Denmark. On this occasion the European Health Report 2002 was released. This report confirms the strong links between socioeconomic development, health and equity in the WHO European Region which covers 51 WHO Member States with some 870 million people. While overall levels of health in the Region are among the highest in the world, the report describes widening gaps between and within countries (www.euro.who.int/europeanhealthreport). PMID- 12631996 TI - Distance learning: an introduction from the guest editors. PMID- 12631997 TI - A multimedia case based approach to the study of office ergonomics. AB - Multimedia technology has the capacity to provide students with an interactive approach to problem based learning and to bridge the gap between theoretical knowledge and clinical practice. The present paper describes the design and development of the program ErgoROM, a CD which presents a case study to assist occupational therapy students in their studies of office ergonomics. A pilot study showed that overall, 91% of respondents rated the ErgoROM as either "Excellent" or "Very Good". Additionally they reported that ErgoROM had a positive impact on active learning and critical thinking. PMID- 12631998 TI - Evaluation of a web-based rheumatology course for students in rehabilitation sciences. PMID- 12631999 TI - Application of multimedia in CAI for rehabilitation education. PMID- 12632000 TI - Learning technologies at the service of higher education: global trends and local Israeli opportunities. AB - This paper provides an overview of global trends in e-Learning and opportunities opened by harnessing learning technologies at the service of the Israeli higher education. The introduction of innovative learning technologies is examined in light of an organizational change. A series of substantial issues should be taken into account to successfully manage the change. Among these are: rethinking of academic learning and instruction, redefining the role of the teachers and the students, decoupling the learning materials and the learning process, new approaches to student assessment and program evaluation, and instituting a well thought out organizational plan including a support center. The above-mentioned issues guided the introduction of a recent call for project proposals for the integration of technologies into Israeli higher education. The main principles, criteria and results of this call are described. PMID- 12632002 TI - Employment and disability in the United Kingdom: an outline of recent legislative and policy changes. AB - In 1996, a paper (Floyd, 1996), on the Vocational Rehabilitation Services in the United Kingdom, reviewed the way in which the services had evolved during the past 50 years, since the end of the second world war. The author described the Disabled Persons (Employment) Act 1944 and the development of rehabilitation and placement services and sheltered workshops over the years. He also gave readers a glimpse of possible future changes, and, in particular, speculated that the UK might follow the United States in the professional training and development of employment and vocational rehabilitation personnel. This paper describes first the main services currently provided by Government and then outlines the quite significant legislative changes and one major policy development that have taken place in the past five years. This includes an overview of the Disability Discrimination Act 1995 which came into force in late 1996, a brief look at some of the early outcomes, and describes the latest proposals for amendment. A description of the major policy development (New Deal for Disabled People) is also given but it is too soon to report on its effectiveness. Whether the changes will lead to any lasting improvement in labour market participation by disabled people, remains an unanswered question; certainly, up to the present, from a statistical point of view, it seems that the position remains much the same (Curtis J, forthcoming). The paper concludes with discussion of the latest key issues and returns to the question raised in 1996 about the training of employment and rehabilitation personnel. PMID- 12632001 TI - Management commitment to safety and health in residential construction: HomeSafe spending trends 1991-1999. AB - The support of good management is fundamental to the success of any safety and health program. Residential construction is a high-risk industry requiring significant commitment by management to impact day-to-day safety and health challenges. Investigators have evaluated management practices and spending trends in a cohort of 228 residential homebuilders in the Denver metro area of Colorado. Findings suggest that companies significantly increased dollars allocated to support safety and health practices between 1991 and 1999. In addition, the HomeSafe Pilot Program has positively impacted financial commitments of partner companies. Resource allocations were significantly greater for specific expense categories when comparing pre to post HomeSafe intervention. This paper presents data on the use of written safety and health programs, safety committees, and workers compensation premium cost containment certification, as well as allocations to safety incentive programs (SIP), personal protective equipment (PPE), other safety equipment (OSE), and safety training (ST). PMID- 12632003 TI - The nature of working in the United States: an occupational therapy perspective. AB - Although working is considered by occupational therapy practitioners to be an important life role, there is very little information about the nature of working in the occupational therapy literature. This article reviews the construct of working in three ways: 1) The history of working and how it has shaped the modern beliefs about working; 2) The nature of working; and 3) The health promoting attributes of working. The paper will also discuss research done on how working is viewed by individuals with disabilities and will touch on some of the implications of the cultural bias in the workplace toward workers with disabilities. Through this increased knowledge of the nature of working, occupational therapy practitioners and other vocational rehabilitation professionals will be better equipped to break down the subtle cultural barriers, and help individuals with disabilities to participate in working. PMID- 12632004 TI - Attributes of excellence in work-related assessments. AB - In order for clinicians to select and conduct work-related assessments that demonstrate 'best practice', it is necessary to identify assessments that are considered 'excellent'. OBJECTIVES: The aims of this study were to determine (1) the attributes associated with excellence for each of 3 types of work-related assessment (WPAs, FCEJs & FCENJs), and (2) the differences between the attributes associated with each type of work-related assessment. STUDY DESIGN: A questionnaire was sent to all accredited occupational or vocational rehabilitation providers in Australia, targeting occupational therapists and physiotherapists who conducted work-related assessments. The response rate was 25.3%, and 132 questionnaires were analysed. RESULTS: A MANOVA revealed that the perceived importance of 18 attributes was different between the 3 forms of work related assessment F36, 364=6.54; p<0.001). There was a core of 7 attributes that showed no difference between assessments (accurate, comprehensive, credible, flexible, practical, safe and useful). Two attributes (generalisable and specific) were different for all 3 forms of assessment. A large group of attributes (consistent, measurable, objective, reliable, reproducible, standardised, structured and valid) demonstrated significant differences between WPAs and both forms of FCE. Relevant demonstrated significant differences between FCENJs and both WPAs and FCEJs. Reliability analysis revealed 2 constructs (dependability and utility). CONCLUSIONS: This study demonstrated that there were significant differences between 3 forms of work-related assessments (WPAs, FCEJs & FCENJs) in terms of the attributes considered necessary for excellence for each form of assessment. The attributes were considered to relate to either qualitative or quantitative features of assessment. These features were reflected in the constructs utility, which was comprised of qualitative attributes and associated with broad concepts of validity, and dependability, which was comprised of quantitative attributes and reflected concepts of reliability. There appeared to be a continuum of work-related assessment that ranged from WPAs demonstrating the most qualitative attributes to FCENJs demonstrating the most quantitative features. PMID- 12632005 TI - An analysis of VDT monitor placement and daily hours of use for female bifocal users. AB - A population of 72 bifocal wearers was studied to determine the relationship between VDT (video display terminal) placement for those who reported musculoskeletal pain and those who did not. The mean hours worked was 50.4 minutes (p=0.003) greater for those who reported head/neck pain versus those who did not and 48.6 minutes (p=0.004) greater for those who reported shoulder/arm pain that those who did not. There was no statistically significant difference between the means of monitor height, distance, or angle for those who reported pain symptoms versus those who did not. This study indicates that, self reported pain symptoms are correlated with hours of VDT work and that there appears to be a threshold at approximately five hours for pain symptoms among bifocal wearers. No such correlation can be made for monitor placement PMID- 12632007 TI - The microrheological changes in the course of erythrocyte senescence after phenylhydrazine injection. AB - To study the microrheological characteristics of RBCs during erythrocyte senescence in vivo, an anemia model of rabbit induced by phenylhydrazine injection was developed. Measurements of the hematocrit, the deformation indexes, the blood viscosity and the sedimentation, etc. were performed in vivo for more than 60 days in the processes of RBC senescence. Obvious changes in the RBC's rheological characteristics were found in this senescent model. Compared with our previously developed Wen's model [1,2] in which the entire RBC population was nearly synchronously produced following the induction of spherocytic anemia in the rabbit with antibody serum, the changes of RBC microrheological characteristics for this model showed approximately the same tendency, although Wen's model with antibody serum was much better in its ability to simulation of the nearly normal physiological conditions than the present one with phenylhydrazine injection. Hence, the present model can serve as a model of RBC senescence under abnormal physiological conditions. PMID- 12632008 TI - Effect of hormone replacement therapy upon haemorheological variables. AB - It is already admitted that hormone replacement therapy (HRT) decreases the risk of developing cardiovascular disease, although its mechanism is not clear yet. In the present work, the effect of the HRT upon cellular and plasmatic haemorheological factors determining blood flow properties: blood viscosity, plasma viscosity, plasma fibrinogen, rigidity erythrocyte index and erythrocyte aggregation rate was studied. Menopausal women were followed through a whole year of HTR. Results demonstrate that after six months of treatment there is a diminution in relative blood viscosity and erythrocyte rigidity, with constant values along the second semester. Erythrocyte aggregation, plasmatic and blood viscosity diminution observed during the treatment can be explained by the simultaneous plasma fibrinogen decrease. Modified cellular and plasmatic rheology could produce beneficial effects on blood flow, particularly in microcirculation, presenting a possible mechanism by which HTR decreases the risk of cardiovascular disease development during menopause. PMID- 12632009 TI - Study of erythrocyte shape changes in schistosomiasis. AB - The aim of this work was to study the red cell morphology in different stages of schistosomiasis. Patients were divided into three groups according to the stage of the disease. For each patient, complete clinical examination, liver function tests, renal function tests, complete blood picture, scanning electron microscopy for erythrocytes (SEM) and abdominal ultrasonography were done. Abnormal morphologic changes of a total discoid flat cells of 70.14%, margin changes of 12.34% and 3.55% of cup forms were found. To our knowledge, these marked changes were not reported in any other disease. No statistical differences were found between red cell shape changes and stage of liver disease. There was a positive correlation between portal vein diameter and percentage of flat discoid forms and a negative correlation between surface changes and clinical stage of liver disease. These changes are known to be accompanied by reduction of red cell deformability and impaired capillary flow. PMID- 12632011 TI - Effects of hydration on blood rheology. AB - This study investigated the impact of oral fluid intake on blood rheology of 17 healthy adults following a 12-14 hour overnight fast from food and drink. An oral fluid load of 500 ml was consumed every 30 minutes for 2 hours. Blood viscosity values at shear rates of 1, 10 and 100 s(-1) were reduced (p<0.05 to p<0.01) at 30 and 120 minutes following hydration; however, these differences were not significant after hematocrit correction. With fluid intake, both uncorrected and corrected viscous component of blood viscoelasticity at oscillatory shear rate of 1 s(-1) and at a constant frequency of 2 Hz were reduced (p<0.05 to p<0.001) at all time points as compared to fasting values. The corrected elastic component of blood viscoelasticity increased 90 minutes after hydration (p<0.05). An overnight fast is accompanied by rheological abnormalities that are altered by fluid intake. PMID- 12632010 TI - Oxidative stress and hemorheological changes induced by acute treadmill exercise. AB - The present investigation was designed to evaluate the acute effect of aerobic exercise on oxidative stress and the flow properties of the blood. Fourteen clinically healthy subjects (7 men and 7 women aged 56+/-19 yr) underwent maximal treadmill exercise with blood samples drawn prior to and immediately after exercise. Post-exercise significant increases were observed in plasma lipid hydroperoxides from 6.5+/-2.0 microM to 7.9+/-1.9 microM (p<0.0001) and the relative concentration of plasma fluorescent products associated with red cell peroxidation from 138+/-28 RF to 220+/-92 RF (p<0.005). After exercise there was a rise in the hematocrit from 41.4+/-3.7% to 44.4+/-4.1% (p<0.0001), increases in whole blood viscosity at shear rates of 22.5/sec to 450/sec (p<0.0005), an increase in plasma viscosity from 1.27+/-0.12 cP to 1.36+/-0.11 cP (p<0.01), an increase in red cell rigidity from 2.44+/-0.48 cP to 2.62+/-0.42 cP (p<0.001) and a decrease in erythrocyte sedimentation rate from 26.9+/-18.6 mm/h to 22.5+/-15.9 mm/h (p<0.01). The findings suggest that acute aerobic exercise induces oxidative damage to red blood cells and adversely affects rheological properties of the peripheral blood. PMID- 12632012 TI - A comparative study of hemorheological parameters in transient ischemic attack and acute ischemic stroke patients: possible predictive value. AB - A complete neurological and laboratory assessment was made of 31 transient ischemic attack (TIA) and 33 acute ischemic stroke patients. Laboratory parameters were compared with 33 age- and sex-matched controls. Erythrocyte deformability was characterised by determining the relative cell transit time (RCTT) with a St. George filtrometer. Plasma viscosity was measured with a Haake microviscosimeter. In comparison with controls, fibrinogen content, erythrocyte sedimentation rate (ESR), platelet and leukocyte count, erythrocyte RCTT and plasma viscosity were found to be significantly higher in stroke patients. In TIA patients the elevation of these values was not significant with the exception of platelet count. Our results suggest that the hemorheological alterations observed in TIA and stroke are largely non-specific findings and associated with the atherosclerotic disease of patients. The significant elevation of leukocytes, fibrinogen and plasma viscosity in acute stroke versus TIA probably reflects the systemic acute phase response of organism to cerebral infarction. PMID- 12632013 TI - Cerebral microvascular responses to air embolism-reperfusion in the cat using fluorescence videomicroscopy. AB - Cerebral microvascular changes due to air embolism-reperfusion in the cat were investigated. Air embolism-reperfusion was produced in the cerebral microvessels by an intra-carotid injection of air (0.2-0.3 ml). Air emboli in the cerebral arterioles were observed continuously from the air injection to reperfusion using fluorescence videomicroscopy. Arteriolar diameter was measured based on video images of arterioles filled with rhodamine-B isothiocyanate dextran, and red cell velocity was measured using a dual window technique with FITC-labeled red cells. Air bubbles ceased to move in the arterioles of 20-70 microm diameter and blood flow was almost stopped in distal arterioles. The air emboli were of cylindrical shape in the arterioles, with hemispherical end caps. The emboli progressed slowly at rates of 7-73 microm/sec and then flowed away. The air emboli induced ischemia-reperfusion with the ischemic duration of approximately 1 minute (10 sec 3 min) at the arteriolar level. Cerebral arterioles began to dilate after the formation of the air emboli and significantly dilated by approximately 50% after reperfusion. A transient increase in red cell velocity was observed after reperfusion. Arteriolar dilation and the increase in velocity led to a hyperemic response in arteriolar flow to ischemia-reperfusion. PMID- 12632014 TI - Analysis of ploidy in hypopharyngeal cancer by laser scanning cytometry on fine needle aspirate biopsies. AB - AIM: To test laser scanning cytometry (LSC) for the analysis of ploidy in squamous cell carcinoma of the hypopharynx (SCCH) and to develop a routine application for minimal samples such as fine needle aspirate biopsies (FNABs). METHODS: From 11 individuals 30 FNABs of primary tumors (n=11) and lymphatic metastases of SCCH (n=11) and non-metastatic lymph nodes (n=8) are analyzed by LSC. This microscope based instrument scans the cells after immobilization on a glass slide and after double staining of cytokeratin and DNA. The location of each cell is stored with the fluorescence data. Therefore the morphology of every cell can be documented by re-staining with H & E; and re-localization on the slide. Additionally, aliquots are Feulgen-stained for image cytometry in 8 specimens. RESULTS: The diploid reference peak is identified taking leukocytes as internal standard. The DNA-index of the carcinoma cells ranges from 0.4 to 3.8. Comparison with image cytometry shows good correlation (r=0.89). CONCLUSION: LSC provides a reliable and objective way to determine the ploidy of SCCH pre operatively. Colour figures can be viewed on http://www.esacp.org/acp/2003/25 2/gerstner.htm. PMID- 12632015 TI - An original approach for quantification of blood vessels on the whole tumour section. AB - Relative abundance of tumour angiogenesis has been shown to be of clinical relevance in cancers of various locations such as the ovary. Nevertheless, several problems are encountered when quantifying tumour microvessels: (i) as many other tumour markers, vascularity pattern is often heterogeneous within the tumour mass and even within the same histological section. As a consequence, an adequate acquisition method must be developed for accurate field sampling. (ii) Manual microvessel counting is long, tedious and subject to poor reproducibility. Introduction in routine practice requires a fast, reproducible and reliable automatic image processing. In this study we present an original procedure combining a slide scanner image acquisition and a fully automatic image analysis sequence. The slide scanner offers the advantage of recording an image of the whole histological section for subsequent automatic blood vessel detection and hot spot area location. Microvessel density and surface fraction were measured for the whole section as well as within hot spots. Different immunostaining methods were tested in order to optimise the procedure. Moreover, the method proposed was submitted to a quality control procedure, with reference to interactive identification of microvessels at scanner level. This experiment showed that 93 to 97% of blood vessels were detected, according to the staining protocol used. Colour figures can be viewed on http://www.esacp.org/acp/2003/25 2/kim.htm. PMID- 12632016 TI - Expression of the cell adhesion molecule CD146/MCAM in non-small cell lung cancer. AB - Expression of MCAM is observed in a variety of human malignancies. We aimed to determine the rate of MCAM expression in our non-small cell lung cancer (NSCLC) collection and to clarify its correlation with clinicopathological parameters and patient survival. 85 NSCLC were analysed immunohistochemically using a monoclonal MCAM antibody (clone N1238) on an NSCLC tissue micro array. The staining was semiquantitatively scored. We found MCAM expression in 51% of NSCLC, preferentially squamous cell carcinomas (p=0.004). No other correlations to tumour size, grade, or stage were found. Univariate survival analysis showed no significant differences of MCAM positive and negative tumours. In adenocarcinomas however, MCAM positivity was significantly associated with shorter patient survival (p=0.016). We conclude, that MCAM is expressed in a high proportion of NSCLC and might be predictive of shortened patient survival in adenocarcinomas of the lung. Colour figure can be viewed on http://www.esacp.org/acp/2003/25 2/kristiansen.htm PMID- 12632018 TI - Gain in 1q is a common abnormality in phyllodes tumours of the breast. AB - We studied DNA copy number changes by CGH and allelic imbalance (AI) on 3p by LOH analysis on 22 phyllodes tumours (PT) of the breast in order to gain insight into the genetic basis of tumour progression in PT. Copy number changes were observed in 14 cases (63%). Gain in 1q with 1q21-23 as the minimal overlapping area was seen in 12 cases (55%). The gain was observed both in benign and malignant tumours. Our study did not reveal any DNA copy number changes or allelic loss on 3p. The results suggest that DNA copy number changes are not associated with the histological grade or clinical behaviour of PT and the chromosomal changes on 3p appear to be rare. Colour figure can be viewed on http://www.esacp.org/acp/2003/25-2/jee.htm PMID- 12632017 TI - DNA extraction from bronchial aspirates for molecular cytology: which method to take? AB - OBJECTIVE: To date, there are only few systematic reports on the quality of DNA extracted from routine diagnostic cytologic specimens. It was the aim of the present study to evaluate the ability of 50% ethanol/2% carbowax (Saccomanno fixative) to preserve bronchial secretions with high quality genomic DNA as well as to compare different DNA extraction methods. METHODS: DNA was extracted from 45 bronchial aspirates by four different extraction protocols. Beside DNA yield, DNA quality with regard to purity, integrity, and PCR success rate were investigated. RESULTS: No fragmentation of sample DNA due to the fixative was detected. It was preserved as high molecular weight DNA. DNA yield, purity, and integrity were dependent on the DNA extraction method to some extend. Irrespective of the DNA extraction method the PCR success rate for amplification of beta-globin gene fragments (268, 536, and 989 bp) was 100%. CONCLUSION: A fixative containing 50% ethanol/2% carbowax preserves high quality DNA which is well suited for PCR-based assays regardless of the extraction protocol used. The selection of the DNA extraction protocol has to be adjusted to the circumstances of application. PMID- 12632019 TI - In vitro model for studying malignancy associated changes. AB - Malignancy associated changes (MAC) can be defined as subtle morphological and physiologic changes that are found in ostensibly normal cells of patients harboring malignant disease. It has been postulated that MAC have a potential to become a useful tool in detection, diagnosis and prognosis of malignant diseases. An in vitro cell culture model system was designed to study interactions between non-small cell lung cancer (NSCLC) and the normal bronchial epithelium of the human respiratory tract in vivo to see if the MAC-like phenomenon can be detected in such a system. In this study we examined changes in nuclear features of normal human bronchial epithelial cells (NHBE) when they were co-cultured with cells derived from a lung cancer cell line NCI-H460. Using discriminant function analysis, nuclear features were determined which allow maximal discrimination between normal cells incubated with or without cancerous cells. Our results demonstrate that MAC appear to be specific to changes induced by malignancy, and that these changes differ from those induced by growth factors in the serum. This study provides evidence in support to the hypothesis that MAC are induced by a soluble factor(s) released by malignant cells. Colour figure can be viewed on http://www.esacp.org/acp/2003/25-2/sun.htm PMID- 12632020 TI - Factor V Leiden mutation and PAI-1 gene 4G/5G genotype in thrombotic patients with Behcet's disease. AB - Behcet's disease is a chronic systemic vasculitis with particular systemic features including thrombotic events. The present study was designed to analyse the role of the factor V Leiden and the prothrombin G20210A mutations and plasminogen activator inhibitor-1 4G/5G polymorphism on the thrombotic risk of patients with Behcet's disease. A total of 50 unrelated patients with Behcet's disease (34 male, 16 female) were the subjects of the study. Twenty-seven of 50 patients with a history of thrombosis comprised group 1, and 23 patients with no thrombosis comprised group 2. In group 1, nine of the 27 patients (33%) were found to have the factor V Leiden mutation (7.1% in healthy population), and the 4G/4G genotype was found in 23% of the patients (26% in control). No patient had the prothrombin G20210A mutation (2.2% in healthy control). In group 2, two patients (9%) had the factor V Leiden and one patient (4%) had the prothrombin G20210A mutations. The 4G/4G polymorphism was found in 30.5% of the patients. The differences in the frequencies of factor V Leiden mutation between group 1 and group 2 (odds ratio, 5.3; 95% confidence interval, 1.0-27.5) and between group 1 and the healthy population were statistically significant ( 0.05). No statistically significant association was found for the prothrombin G20210A mutation and the 4G/5G genotype between the two groups or between each group and the healthy population, indicating that the prothrombin G20210A mutation and the 4G/4G polymorphism do not play a role in the development of thrombosis in Behcet's disease. These data suggested that the factor V Leiden mutation might be a risk factor for the development of thrombosis in Behcet's disease patients. PMID- 12632021 TI - Circulating tissue factor, tissue factor pathway inhibitor and D-dimer in umbilical cord blood of normal term neonates and adult plasma. AB - The investigation of many hemostatic defects in newborns is restricted by the lack of normal reference values. The coagulation system of the neonate differs in many ways from that of the adult. The present study was designed to compare the concentrations of tissue factor (TF), tissue factor pathway inhibitor (TFPI) and D-dimer (DD) in the umbilical cord blood of healthy newborns and in adult plasma. TF antigen was quantified using an in-house enzyme-linked immunosorbent assay, whereas TFPI and DD levels were measured with commercial kits. The mean TF level in cord blood (mean standard deviation, 183.94 103.63 pg/ml) was significantly higher ( = 0.008) than that in adults (136.64 65.09 pg/ml). Cord blood exhibited enhanced fibrinolysis, as was reflected by a significantly higher level of DD (924.57 733.87 ng/ml, 0.001) than that in adults (45.57 17.21 ng/ml). Conversely, cord blood (30.88 10.16 ng/ml) demonstrated significantly lower ( 0.001) TFPI levels than that in adults (55.77 21.16 ng/ml). However, no significant differences of these three hemostatic markers were noted between both gender groups in newborns and adults. Our findings indicate that an active and dynamic state of hemostasis exists in cord blood, as the fluidity of cord blood remains preserved in the presence of birth injury. PMID- 12632022 TI - Activation of calcium-dependent calmodulin by calcium(II)3(3,5 diisopropylsalicylate)6(H2O)6 decreases thrombin receptor activating peptide induced P-selectin expression. AB - We examined the influence of 3,5-diisopropylsalicylic acid (3,5-DIPS) and calcium(II)3 (3,5-diisopropylsalicylate)6 (H2 O)6 [Ca(II)3 (3,5-DIPS)6 ], a new activator of calcium-dependent calmodulin-triggered nitric oxide synthase, on thrombin-induced platelet P-selectin expression. Citrated whole blood samples were incubated with either ethanol vehicle, 3,5-DIPS, or Ca(II)3 (3,5-DIPS)6. These whole blood samples were also co-incubated with thrombin receptor activating peptide (TRAP) or adenosine diphosphate (ADP), to up-regulate P selectin (CD62P) on platelets. Both TRAP and ADP up-regulated P-selectin on platelets compared with platelets in whole blood samples that were not incubated with either platelet activator. Co-incubation of whole blood samples with TRAP, ADP together with 3,5-DIPS, or Ca(II)3 (3,5-DIPS)6 revealed that Ca(II)3 (3,5 DIPS)6 caused a decrease in platelet P-selectin expression for TRAP, ADP, and no activator co-incubated samples of whole blood. Incubation of platelets with 3,5 DIPS also caused a decrease in ADP-induced up-regulation of P-selectin but failed to affect TRAP or no-activator-treated platelets. Incubation of whole blood with Ca(II)3 (3,5-DIPS)6 induced some hemolysis. We found that hemolysis increases basal P-selectin expression on platelets. We therefore conclude that Ca(II)3 (3,5 DIPS)6 decreased not only basal, but also hemolysis-induced P-selectin expression on platelets. In contrast, incubation of haemolysed whole blood with SIN-1 (standard nitric oxide-releasing drug) had no effect on P-selectin expression. In summary, Ca(II)3 (3,5-DIPS)6, a new calmodulin-dependent nitric oxide synthase activator, decreases P-selectin expression of human platelets in response to thrombin receptor activation. Improved calcium-dependent calmodulin activators may become useful drugs for the treatment of disorders associated with platelet activation, and P-selectin may decrease expression due to hemolysis. PMID- 12632023 TI - Inhibition of endothelial cell-mediated generation of activated protein C by direct and antithrombin-dependent thrombin inhibitors. AB - The present study investigated the effect of the thrombin inhibitors antithrombin (AT) (with and without unfractionated heparin or low molecular weight heparin), hirudin, inogatran and melagatran on thrombin-thrombomodulin-mediated generation of activated protein C (APC), in solution and on endothelial cells. Sequential incubation with thrombin, thrombin inhibitors and protein C was followed by measurement of APC by an amidolytic assay. The approximate concentrations resulting in 50% inhibition of endothelial cell-mediated APC generation for AT, AT-unfractionated heparin, AT-low molecular weight heparin, hirudin, melagatran and inogatran were 200, 4, 9, 1, 8 and 60 nmol/l, respectively. The normal plasma level of AT is 2800 nmol/l and relevant therapeutic concentrations from clinical trials are 200 nmol/l for hirudin, 500 nmol/l for melagatran and 1000 nmol/l for inogatran. The present study indicates that clinically relevant concentrations of the tested thrombin inhibitors interfere with endothelial-mediated APC generation, which may offer an explanation for the lack of a dose-response effect in clinical trials with thrombin inhibitors. PMID- 12632024 TI - Fibrinolytic activity and bleeding after cardiac surgery with cardiopulmonary bypass and low-dose aprotinin therapy. AB - Low-dose aprotinin inhibits hyperfibrinolysis in cardiac surgery. However, excessive postoperative bleeding and increased fibrinolysis may occur despite low dose-aprotinin administration. We investigated (i) whether fibrinolytic activity significantly rises under low-dose aprotinin administration, and (ii) whether this is associated with excessive postoperative bleeding (> or = 1000 ml/24 h). In a prospective single-blind trial, 120 consecutive patients were randomized to receive 280 mg aprotinin or no aprotinin before skin incision. D-dimer levels increased significantly to the end of surgery, reaching higher levels in the control group. The risk for excessive bleeding was lower in the aprotinin group (12 versus 37%, = 0.001). Fifteen minutes after heparin reversal, patients were at risk for excessive bleeding, when enhanced fibrinolysis was documented (aprotinin group, D-dimer > or = 1.0 micro g/ml, odds ratio = 9.1, = 0.047; control group, D-dimer > or = 3.0 micro g/ml, odds ratio = 4.6, = 0.014). Ninety seven per cent of the aprotinin group and 81% of control group patients had no excessive bleeding when the D-dimer plasma level was below these values. We conclude that (i) fibrinolytic activity significantly rises under low-dose aprotinin administration, and (ii) plasma D-dimer levels at end of surgery may help to identify patients who are unlikely to develop excessive postoperative bleeding. PMID- 12632025 TI - The G-to-T point mutation in codon 34 of the factor XIII gene and the risk of pre eclampsia. AB - A G-to-T point mutation in exon 2 of the FXIII A-subunit gene results in a leucine rather than valine at amino acid position 34 of the factor XIII molecule. The presence of leucine has been associated with a reduced risk of both arterial and venous thrombosis. We examined the prevalence of this point mutation in 102 cases of pre-eclampsia and 208 matched control subjects, as inherited and acquired risk factors for arterial and venous thrombosis are associated with an increased risk of pre-eclampsia. The GT genotype was observed in 38% of controls and 29.4% of cases and the TT genotype was observed in 6.7% of controls and 5.9% of cases. In subjects heterozygous for the T genotype (GT) the relative risk of pre-eclampsia was 0.7 [95% confidence interval (CI95 ) 0.4-1.1] when compared with the GG genotype. For subjects homozygous for the T allele the relative risk for pre-eclampsia was 0.8 (CI95 0.3-2.1) when compared with the GG genotype. The risk associated with the T allele in the heterozygous and homozygous forms compared with the GG genotype was 0.7 (CI95 0.4-1.1). We conclude that the presence of leucine at this site is not associated with a protection against pre eclampsia to the magnitude of that reported in other thrombotic disease. PMID- 12632026 TI - Thrombin-induced platelet activation and its inhibition by anticoagulants with different modes of action. AB - Thrombin-induced platelet activation involves cleavage of protease-activated receptors (PARs) 1 and 4, and interaction, via glycoprotein (Gp)Ibalpha, with the platelet GpIb/IX/V complex. This study investigated inhibition of platelet activation by thrombin inhibitors with different modes of action: two reversible direct thrombin inhibitors, melagatran and inogatran; hirudin, a tightly binding direct thrombin inhibitor; and two indirect thrombin inhibitors, heparin and dalteparin. Up-regulation of P-selectin (CD62P) and PAR-1 cleavage was measured in human whole blood, by flow cytometry. The thrombin concentration that induced 50% of maximum (EC50 ) PAR-1 cleavage was 0.028 nmol/l, while that of platelet activation (CD62P) was over two-fold higher (0.64 nmol/l). The EC50 of a PAR-1 independent component, defined as a further activating effect of thrombin on top of the maximum PAR-1-activating peptide (AP) effect, was 3.2 nmol/l. All anticoagulants were concentration-dependent inhibitors of thrombin-induced platelet activation and PAR-1 cleavage, but none inhibited PAR-1-AP or PAR-4-AP induced activation. Melagatran and inogatran were more potent inhibitors of CD62P up-regulation than of PAR-1 cleavage; conversely, hirudin, heparin and dalteparin were more potent inhibitors of PAR-1 cleavage.Thus, reversible direct thrombin inhibitors, such as melagatran, are potent inhibitors of thrombin-induced platelet activation, acting mainly by inhibition of a PAR-1-independent component. PMID- 12632027 TI - Recombinant factor VIIa in orthotopic liver transplantation: influence on parameters of coagulation and fibrinolysis. AB - The effect of recombinant factor VIIa (rFVIIa) on blood loss was evaluated in cirrhotic patients undergoing orthotopic liver transplantation. In the present study, we explored the effect of rFVIIa on coagulation and fibrinolysis during orthotopic liver transplantation. Coagulation factors, parameters of thrombin generation and parameters of fibrinolysis were measured in six patients who had received a single dose of 80 micro g/kg rFVIIa and in ten controls, during and after orthotopic liver transplantation. Baseline concentrations and course of coagulation factors were similar in patients and controls. Thrombin generation did not rise after the administration of rFVIIa, but showed a sharp increase after reperfusion in patients, as compared with controls. No difference in fibrinolysis was apparent between patients and controls. No evidence of diffuse intravascular coagulation was seen. We conclude that the use of rFVIIa in orthotopic liver transplantation seems to enhance thrombin generation in a localized and time-limited matter, without causing systemic coagulation. PMID- 12632028 TI - Correlation between thrombin potential and bleeding after cardiac surgery in adults. AB - We used a sensitive assay to measure thrombin potential in 20 patients who underwent cardiopulmonary bypass surgery for coronary artery bypass grafts. We measured coagulation factors II, V, VII, VIII and X. Blood loss was measured as the total amount in the mediastinal drains in the first 24 h postoperatively. Thrombin potential was median 107 nmol/l.min (range 62-181) preoperatively and median 46 nmol/l.min (range 19-120) postoperatively. Coagulation factors II, V, VII,VIII and X were within normal limits preoperatively. Factor II fell from 77 IU/dl preoperatively to 37 IU/dl at 120 min postoperatively. Factor V fell from 85 IU/dl preoperatively to 61 IU/dl postoperatively. Factor VII fell from 91 IU/dl to 66 IU/dl postoperatively. Factor VIII was 128 IU/dl preoperatively and 127 IU/dl postoperatively. Factor X fell from 90 IU/dl preoperatively to 50 IU/dl postoperatively. Total blood loss in 24 h in the mediastinal drains postoperatively was mean 673 ml, median 650 ml (range 250-2000). Reduction in thrombin potential correlated inversely with postoperative blood loss, r= -0.75 (Spearman correlation). The fall in the thrombin potential correlated with the prothrombin level (r = 0.75) and factor X (r = 0.47). PMID- 12632029 TI - Fibrinolytic response to venous occlusion is decreased in patients after Kawasaki disease. AB - Impaired fibrinolysis is considered a sensitive marker of endothelial dysfunction. Persistent endothelial dysfunction occurs in some patients following Kawasaki disease. The aim of the present study was to assess whether impaired fibrinolysis is present in long-term survivors of Kawasaki disease. The study included 42 children with a documented history of Kawasaki disease presenting with or without coronary lesions, and 26 healthy controls. Blood samples were collected from patients and controls prior to and following venous occlusion stress testing. Significantly decreased fibrinolytic response to venous occlusion was detected in patients compared with controls due to decreased tissue plasminogen activator. In addition, patients had significantly increased plasma concentrations of plasminogen and fibrinogen, which were related to similar increases of alpha2 -macroglobulin. Decreased fibrinolytic response was found in patients with coronary aneurysms but also in those without coronary lesions. In summary, a decreased fibrinolytic response to venous occlusion may reflect persistent endothelial damage following acute Kawasaki disease, potentially predisposing these patients to accelerated atherosclerosis and cardiovascular disease in early adult life. PMID- 12632030 TI - Recombinant activated factor VII combined with local measures in preventing bleeding from invasive dental procedures in patients with Glanzmann thrombasthenia. AB - Recombinant activated factor VII (rFVIIa), combined with local measures of fibrin glue and a celluloid splint, preventing bleeding from four invasive dental procedures is reported. A single dose of 180-200 micro g/kg was successfully used in three surgical removals of impacted teeth. Four doses of rFVIIa were required in another full mouth treatment of extraction, pulpotomy, filling and the stainless steel crowning of 13 teeth. The repeated dose of rFVIIa was given whenever the bleeding complication was visualized. It is cost-effective for preventing external bleeding. Additionally, an oral rinsing solution of tranexamic acid (25 mg/kg) was given three times a day for 7 days. In conclusion, rFVIIa has been shown to be an effective alternative to platelet concentrate in patients with Glanzmann thrombasthenia. PMID- 12632031 TI - Familial thrombophilia is an oligogenetic disease: involvement of the prothrombin G20210A, PROC and PROS gene mutations. AB - Population-based case-control studies and cases previously published suggest that the prothrombin G20210A mutation is a weak risk factor for thrombosis, leading to clinical expression mainly in the presence of other risk factors. We report the results of plasma and genetic analyses performed in a 13-year-old symptomatic boy homozygous for the 20210A allele and in his family, which are in accordance with this suggestion. These analyses demonstrated the presence of several PROC (R-5W, R87H) and PROS (R60C, T103N) gene mutations in this family. These additional mutations have modulating effects on clinical expression of the G20210A mutation. The present family study illustrates the concept of 'mild' mutation and the hypothesis that familial thrombophilia is a multifactorial disease. PMID- 12632032 TI - Comparison of plasma fibrinogen by Clauss, prothrombin time-derived, and immunonephelometric assays in a general population: implications for risk stratification by thirds of fibrinogen. AB - There is strong evidence from meta-analyses of prospective epidemiological studies that increasing plasma fibrinogen levels are associated with increasing risk of ischaemic heart disease. It has been suggested that categorization of plasma fibrinogen by thirds of the population distribution be added to cardiovascular risk prediction equations. However, the heterogeneity of plasma fibrinogen and the resulting discrepancies between commonly performed assays may lead to differences in both mean levels and distributions, and in categorizations of populations by thirds. We therefore compared three commonly performed routine fibrinogen assays in a random population sample of 1373 men and women aged 25-64 years in the fourth World Health Organization MONICA survey in north Glasgow. The two assays of clottable fibrinogen (von Clauss and prothrombin time derived) showed similar mean values and distributions, whereas the immunonephelometric assay showed lower mean values. There was significant disagreement between all three assays in categorization of thirds of population fibrinogen distribution (kappa statistic, 0.64 von Clauss versus prothrombin time derived, 0.46 von Clauss versus immunonephelometric, and 0.51 prothrombin time derived versus immunonephelometric). We conclude that further standardization of plasma fibrinogen assays is desirable for ischaemic heart disease risk stratification, and that further studies of the causes and clinical significance of discrepancies between fibrinogen assays in the general population are indicated. PMID- 12632033 TI - Influence of soluble fibrin on reaction kinetics of plasmin type 1 and type 2 with alpha2-antiplasmin. AB - This study investigates, by slow binding kinetics methods, reaction kinetics of both plasmin types 1 and 2 with alpha -antiplasmin in the presence of increasing concentrations of either epsilon-amino-caproic acid (EACA) or soluble fibrin. All curves of plasmin-alpha -antiplasmin interaction followed the same pattern, indicating reversible slow binding inhibition with an initial loose complex and a following tight complex. Without soluble fibrin or EACA, differences between plasmin types 1 and 2 could be seen in the initial loose complex formation. The presence of increasing concentrations of EACA slowed down the first step of the reaction (without any effect on the second step), resulting in increasing values for K. Plasmin type 1 demonstrated a steep slope of K at an EACA concentration of 1 mmol/l. In plasmin type 2, the increase of K started at higher EACA concentrations. The value for K at a high EACA concentration (100 mmol/l) was the same for both plasmin types. In contrast to EACA, increasing concentrations of soluble fibrin slowed down both reaction steps. At high concentrations of soluble fibrin, the inhibitory effect of alpha -antiplasmin was almost completely abolished. Our data demonstrate that the effect of soluble fibrin and the lysine analogue EACA on plasmin-antiplasmin reactions are different and that the use of lysine analogues does not mimic fibrin in laboratory analyses of plasmin inhibition. In addition, our data indicate theoretical differences between plasmin type 1 and plasmin type 2, when used for local thrombolytic therapy.(2) (2) (i initial) (i initial) (i initial) (i initial) (2) PMID- 12632034 TI - Assessment of blood coagulation in severe liver disease using thromboelastography: use of citrate storage versus native blood. AB - Thromboelastography evaluates the viscoelastic properties of blood coagulation. Using native blood, measurement must start soon after sampling. With normal coagulation, native and citrated blood values correlate well. No data exists from cirrhotic patients. We compared native and citrate thromboelastography parameters in 30 cirrhotic patients (20 Child-Pugh C class, two liver failure). Thromboelastography was performed within 4 min using native blood and after recalcification within 1-2 h of citrate storage. Thromboelastography variables (, alpha, ) were compared using the Mann-Whitney test, correlation investigated with the Pearson method and the degree of agreement with the Bland-Altman method. There was no significant difference between citrated and native blood for all variables. Median values for native and citrated were, respectively, 16.4 (range 2.3-22.5) and 15.1 (range 9.8-29.9); 6.3 (range 3.5-11.3) and 6.2 (range 2.8 10.9); 48.3 (range 30.7-62.9) and 46.2 (range 30.4-60.4); angle alpha 30.8 (range 18.7-46.8) and 33.2 (range 19.9-55.8). Correlation for each variable was significant ( 0.01). There was a good degree of agreement for all but two patients (both bleeding) for all variables. Citrated blood can substitute native blood using thromboelastography in cirrhotic patients, allowing more time between sampling and the thromboelastography measurement. PMID- 12632035 TI - Two novel cases of cerebral haemorrhages at the neonatal period associated with inherited factor VII deficiency, one of them revealing a new nonsense mutation (Ser52Stop). AB - Factor VII (FVII) is a plasma glycoprotein that plays a key role in the initiation of blood coagulation cascade. Inherited FVII deficiency is a rare autosomal recessive disorder with a wide heterogeneous clinical pattern. The severe form may be associated with intracranial haemorrhages occurring closely to birth with a high mortality rate. In the present article, we report two novel cases of neonatal intracerebral bleeding associated with FVII activity levels below 1% of normal. FVII genotyping investigations revealed particular genotypes including the deleterious Cys135Arg mutation and a novel Ser52Stop nonsense mutation at the homozygous state. Both mutations, through different mechanisms, are expected to be inconsistent with the production of functional FVII. These putative mechanisms are discussed through a review of the literature on phenotypic and genotypic characteristics of cerebral haemorrhages in severe inherited FVII deficiency. PMID- 12632036 TI - Three-dimensional analysis of mouse rod and cone mitochondrial cristae architecture: bioenergetic and functional implications. AB - PURPOSE: Comparative studies of structure related to function offer a promising means of understanding the significance of differences in cytoarchitecture. Mitochondrial crista structure is linked tightly to mitochondrial function. Non foveal cone photoreceptors of primates contain considerably more inner segment mitochondria and have higher oxidative enzyme activity than do rods. In addition, it is suggested that light-adapted cones have a higher aerobic ATP demand than light-adapted rods. Therefore, we investigated the oxidative metabolism and three dimensional membrane architecture of mouse rod and cone inner segment mitochondria. METHODS: We determined the number, size, cytochrome c oxidase (CO) reactivity, and membrane architecture of rod and middle wavelength-sensitive (M) cone inner segment mitochondria from 21 day old light-adapted C57BL/6 mice using conventional electron microscopy and the three-dimensional approach of single- and double-tilt electron microscope tomography. Fourteen different measurements of mitochondrial substructures were analyzed. Photoreceptor oxygen consumption was determined in dark- and light-adapted retinas. RESULTS: Rod and cone mitochondria displayed an orthodox conformation. Cone inner segments, compared to rods, contained 2-fold more mitochondria and were more CO reactive. Rod and cone mitochondria had similar outer-inner membrane width, contact site width, diameter and density, crista width, number of cristae/volume, number of cristae segments/volume, and fraction of cristae with multiple segments. In contrast, cone mitochondria had narrower crista junctions, greater cristae connectivity, and approximately 3-fold more cristae membrane surface area compared to rods. The increased cristae membrane surface area in cones was accomplished by connecting more cristae segments together, rather than by creating more cristae. CONCLUSIONS: These results demonstrate that middle wavelength (M) cones have a different bioenergetic signature than do rods and suggest that the aerobic ATP demand and production is greater in light-adapted cones than in light-adapted rods. Cones utilize two complimentary strategies to increase their aerobic ATP production: increase the number of mitochondria and increase the cristae surface membrane area. The greater ATP generation by cones may also provide increased protection against metabolic insults and apoptosis compared to rods. PMID- 12632037 TI - An unusual case of meniscal hematoma mimicking a medial meniscal cyst. AB - This is a case report of a so-called meniscal hematoma that mimicked a meniscal cyst. A literature search found no previous report of it before. Treatment with arthroscopic intra-articular decompression alone is enough. We discuss the possibility of its pathogenesis and treatment. PMID- 12632039 TI - Pathologic infrapatellar plica. AB - The fenestra type of infrapatellar plica was noted arthroscopically in a 57-year old patient with flexion contracture of the knee. It was found to be thickened and fibrotic and to have lost its normal elasticity. The pathologic plica was impinged at the intercondylar notch and trochlea, blocking further extension beyond 25 degrees of knee flexion. After excising the plica, the knee gained a further extension of 17 degrees; however, full extension could not be obtained. The knee gained full extension after notchplasty on the roof at 2 months postoperatively. This is the first report of a pathological infrapatellar plica in an older patient. PMID- 12632038 TI - A true intra-articular lipoma of the knee in a girl. AB - A 16-year-old girl presented with a soft-tissue mass in the anterolateral aspect of her right knee. Magnetic resonance imaging revealed an intra-articular tumor arising from the anterior fat pad of the knee. The tumor showed low signal intensity on T1-weighted images and high signal intensity on T2-weighted images. It was resected under arthroscopic guidance. Grossly, the tumor was composed of soft, yellowish adipose tissue. Pathologic examination revealed a collection of mature adipocytes with marked myxoid changes in the matrix. A diagnosis of lipoma was made. True intra-articular lipomas should be distinguished from lipoma arborescens, which is considered to be a reactive process. In the English language literature, we found only 7 cases of true intra-articular lipoma reported previously. The present case is the youngest patient with such a tumor, and all the previous reports are of the tumor in adult patients. Intra-articular lipoma should be considered in the differential diagnosis of intra-articular masses in adolescents. PMID- 12632040 TI - That bloody knee! Dealing with an organized hemarthrosis. AB - The decision to manage an organized hemarthrosis conservatively or surgically is difficult and still controversial, especially in the postoperative patient compared with the trauma patient. We describe a cheap, effective, and readily available technique for the surgeon who elects for a surgical solution to this problem using a Karman catheter (Rocket Medical PLC, Watford, UK) during arthroscopy to complete an irrigation and vacuum circuit. PMID- 12632060 TI - Role of RLIP76 in lung cancer doxorubicin resistance: II. Doxorubicin transport in lung cancer by RLIP76. AB - ATP-dependent transport of doxorubicin (DOX) by recombinant human RLIP76 has been demonstrated previously in reconstituted proteoliposomes. In the preceding communication, we demonstrated that the ATPase activity of RLIP76 was 2-fold higher in NSCLC as compared with SCLC, and it correlated with their inherent DOX resistance. Present studies were performed to determine whether greater ATPase activity of RLIP76 in NSCLC translated into greater RLIP76 mediated DOX transport, and to determine the overall contribution of RLIP76 toward total DOX transport. Consistent with the greater RLIP76 ATPase activity in NSCLC, DOX transport in artificial proteoliposomes reconstituted with purified RLIP76 from NSCLC was 1.8-fold greater than in SCLC. Anti-RLIP76 antibodies completely abrogated DOX transport in these RLIP76 proteoliposomes, whereas anti-MRP or anti Pgp antibodies had no effect on transport. DOX transport studies in crude membrane vesicles from SCLC and NSCLC also showed a 2.1-fold higher rate of transport in NSCLC as compared with SCLC. Anti-RLIP76 IgG, which recognized only RLIP76 in crude extracts of both SCLC and NSCLC, inhibited 67+/-4% (n=12 cell lines) of total DOX transport in crude membrane vesicles from both SCLC and NSCLC. Inhibition of DOX transport by anti-MRP and anti-Pgp antibody was 35+/-7% (n=12) and 2+/-0.3% (n=12), respectively. The mixture of the three antibodies inhibited DOX transport by 95+/-3% (n=12). These studies demonstrate that DOX transport activity of RLIP76 is significantly greater in NSCLC as compared with SCLC, and that RLIP76 is the major DOX transporter in lung cancer cell lines. PMID- 12632061 TI - Role of RLIP76 in lung cancer doxorubicin resistance: III. Anti-RLIP76 antibodies trigger apoptosis in lung cancer cells and synergistically increase doxorubicin cytotoxicity. AB - RLIP76 (ral-binding protein, RalBP1) is a non-ABC multi-specific transporter of amphiphilic chemotherapeutic drugs such as doxorubicin (DOX) and glutathione electrophile conjugates. In the present studies, we used polyclonal rabbit anti human RLIP76 IgG to inhibit RLIP76 function for determining the role of RLIP76 in DOX resistance of NSCLC cells. Western blot analyses and immunohistochemistry studies showed no recognition of other protein in crude NSCLC cell homogenates by anti-RLIP76, confirming the specificity of anti-RLIP76 IgG. In immunohistochemistry and flow cytometry studies, these antibodies recognized RLIP76 domain(s) on the cell surface. Cells coated with anti-RLIP76 IgG accumulated significantly greater DOX than cells coated with pre-immune IgG. Synergy was calculated using the Chou-Talalay median effect analysis. Herceptin was the positive control, and pre-immune IgG and Rituxan (anti-CD20) were negative controls. The interaction of anti-RLIP76 IgG and DOX were markedly synergistic (CI 0.36+/-0.27). Lesser synergy was observed between Herceptin and DOX (CI 0.75+/-0.49). Interaction between Herceptin and anti-RLIP76 was only additive (CI 1.12+/-0.5). Human IgG, Rituxan, and rabbit pre-immune IgG controls had no effect on DOX toxicity. DNA-laddering confirmed that DOX triggered apoptosis. Anti-RLIP76 IgG alone as well as Herceptin alone also triggered apoptosis in all 6 NSCLC cell lines. Anti-RLIP76 IgG and Herceptin were shown to increase DOX accumulation in NSCLC. These results demonstrated that specific inhibition of the transport function of RLIP by anti-RLIP76 IgG can trigger apoptosis and synergistically increase DOX cytotoxicity in NSCLC. PMID- 12632062 TI - The multidrug resistance modulator SDZ-PSC 833 potentiates the photodynamic activity of chlorin e6 independently of P-glycoprotein in multidrug resistant human breast adenocarcinoma cells. AB - Photodynamic therapy has clinical indications in treatment of localized cancers and could be interesting for eradication of local recurrence of chemoresistant tumors. In the present study, the intracellular accumulation and distribution of chlorin e6 was investigated in MCF-7 and in P-glycoprotein overexpressing, doxorubicin resistant MCF-7/DXR cell lines. After 3-h incubation with chlorin e6 (1.7 micro mol.l(-1)), no significant difference in accumulation was observed between MCF-7 and MCF-7/DXR cells. Chlorin e6 cellular efflux did not differ in the two cell lines. The lack of influence of P-glycoprotein was confirmed since SDZ-PSC833 (PSC) had no influence in chlorin e6 accumulation and efflux in MCF 7/DXR cells. The subcellular distribution of chlorin e6 appeared different in MCF 7/DXR than in MCF-7 cells. Double staining colocalization fluorescence microscopy studies were performed to identify the subcellular localization sites for chlorin e6 using organelle probes for endoplasmic reticulum, Golgi apparatus, mitochondria and lysosomes. In MCF-7, chlorin e6 was distributed in all cytoplasmic organelles including endoplasmic reticulum and Golgi. In MCF-7/DXR, a diffuse cytoplasmic distribution was observed excepted for the endoplasmic reticulum and Golgi area in which less chlorin e6 was distributed. In MCF-7/DXR, PSC was found to restore the distribution of chlorin e6 in the endoplasmic reticulum and Golgi area while in MCF-7, no effect on the subcellular distribution of chlorin e6 was observed. Although the photodynamic activity of chlorin e6 (1.7 micro mol.l(-1), 650 nm, 8 mW.cm(-2)) was found to be lower in MCF-7/DXR than in MCF-7 cells, PSC was found to potentiate the photodynamic activity of chlorin e6 to similar extent in both cell lines. These results clearly demonstrate that PSC potentiates the photodynamic activity of Chlorin e6 independently of the expression of P-glycoprotein and further suggest that the photodynamic activity of chlorin e6 could be related to its intracellular distribution in the endoplasmic reticulum and the Golgi. PMID- 12632063 TI - Genome-scale analysis of resveratrol-induced gene expression profile in human ovarian cancer cells using a cDNA microarray. AB - Resveratrol (3,5,4'-trihydroxy-trans-stilbene) is a natural compound found in large quantities, most notably in grapes and red wine, which has been shown to have anti-inflammatory, chemopreventive and anti-angiogenic effects. We examined whether resveratrol has any effect on growth and gene expression in the human ovarian cancer PA-1 cells. We show that resveratrol inhibits cell growth and induces apoptosis in PA-1 human ovarian cancer cells. We also investigated the effect of resveratrol on changes of global gene expression during resveratrol induced growth inhibition and apoptosis in PA-1 cells using a human cDNA microarray with 7,448 sequence-verified clones. Out of the 7,448 genes screened, 118 genes were founded to be affected in their expression levels by more than 2 fold after 24-h treatment with 50 micro M resveratrol. Resveratrol treatment of PA-1 cells at the final concentration of 50 micro M for 6, 12, 24 and 48 h and gene expression patterns were analyzed by microarray. Clustering of the genes modulated more than 2-fold at three of the above times points divided the genes into 2 groups. Within these groups, there were specific subgroups of genes whose expressions were substantially changed at the specified time points. One of the most highly up-regulated genes found in this study was NAD(P)H quinone oxidoreductase 1(NQO-1), which has recently been shown to be involved in p53 regulation. Although the precise roles of genes whose expression levels were found to fluctuate after resveratrol treatment remain to be elucidated, we hope that the new view of gene expression in human ovarian cancer cells following resveratrol exposure, as offered by this study, provides clues for the mechanism of resveratrol action. PMID- 12632064 TI - Nitric oxide mediated photo-induced cell death in human malignant cells. AB - Photodynamic therapy (PDT) is a therapeutic modality used for the treatment of a variety of solid neoplasms. The principle of PDT is based on the selective uptake of a photosensitizing chemical in tumor tissue/cell followed by irradiation of tumors with visible light. The treatment results in a cascade of oxidative events causing cell death both in vitro and in vivo. Nitric oxide (NO) is a gaseous free radical, which is an important modulator of immune, endocrine and neuronal functions and plays an important role in the induction of apoptosis. Hypericin (HY) is a photosensitizing pigment from Hypericum perforatum that displays phototoxic effects in neoplastic cell lines. Our previous studies have shown HY induced apoptotic cell death in nasopharyngeal carcinoma and other tumor cells. To better understand the oxidative mechanism of apoptosis induced by HY, we hypothesized the role of NO in PDT, which is considered to be involved in a variety of physiological and pathological processes. We first demonstrated the presence of nicotinamide adenine dinucleotide hydrogen phosphate-diaphorase (NADPH-d) reactivity, a potential marker of NO synthesizing (NOS) enzyme both at light (LM) and electron microscopic (EM) level. Immunocytochemistry, using specific antibodies for NOS subtypes (constitutive, NOS I and inducible, NOS II), we observed that both NOS I and NOS II was present in all cell lines. The expression of both NOS I and NOS II was further verified using Western blot analysis as early as 15 min post PDT compared to that of drug-treated non irradiated and light alone treated control cells. Our observation of NO production and distribution using the DAF-2 method is direct evidence of NO production in PDT-treated cells. PMID- 12632065 TI - Inhibited proliferation of cyclooxygenase-2 expressing human hepatoma cells by NS 398, a selective COX-2 inhibitor. AB - Hepatocellular carcinoma (HCC) is a growing human health problem worldwide. Limited treatment and poor prognosis of this disease emphasize the importance in developing an effective chemoprevention. Overexpression of cyclooxygenase-2 (COX 2) has been associated with hepatocarcinogenesis. Although COX-2 inhibitors have been tested for chemoprevention of colon cancer, it remains unknown whether these agents possess anti-HCC effects as well. The present study assessed the effects of a selective COX-2 inhibitor, NS-398, on proliferation of human hepatoma cells in association with COX-2 expression, and the possible mechanisms. In four tested human hepatoma cell lines, overexpression of COX-2 was confirmed in HepG2, HuH7, and Chang liver cells, but not in PLC/PRF/5 cells. Addition of 50 micro M NS-398 resulted in both dose-dependent and time-course inhibition of HepG2 proliferation. In contrast, addition of 50 micro M NS-398 to COX-2 non-expressing PLC/PRF/5 cells resulted in only a mild reduction of cell proliferation. Consistent with this, a 48-h culture of HepG2 cells with 50 micro M NS-398 caused a significant decrease of prostaglandin E2 (PGE2) production. While, the same NS 398 treatment showed only a mild suppression of PGE2 production in COX-2 non expressing PLC/PRF/5 cells. These findings indicate that NS-398-induced suppression of HepG2 proliferation appears mediated by decreased COX 2/prostaglandin (PG) production. We also found that NS-398-induced inhibition of HepG2 proliferation was associated with decreased 5-bromo-2'-deoxyuridine (BrdU) uptake, suggesting a reduced cell cycle progression in G1-S transition. NS-398 treatment also enhanced the apoptotic rate in COX-2 expressing HepG2 cells, but not in COX-2 non-expressing PLC/PRF/5 cells. Our findings confirmed an effective inhibitory effect of NS-398 on proliferation of COX-2 expressing human hepatoma cells through a decreased COX-2/PG activity that is associated with altered cell cycle progression and apoptotic rate. PMID- 12632066 TI - Potential involvement of IL-8 and its receptors in the invasiveness of pancreatic cancer cells. AB - The purpose of the present study was to examine the expression of interleukin (IL)-8 and IL-8 receptors and to evaluate the effects of IL-8 on human pancreatic cancer. We examined the expression of IL-8 and its two receptors (CXCR1 and CXCR2) in 40 surgically resected human pancreatic cancer tissues and in three different human pancreatic cancer cell lines (PANC-1, MIAPaCa-2 and Capan-2). The immunohistochemical analysis using specific antibodies demonstrated that positive staining for IL-8, CXCR1 and CXCR2 in surgically resected human pancreatic cancer was 50, 55 and 65%, respectively. Moreover, 40% of these cases were positive for both IL-8 and IL-8 receptors. In contrast, immunoreactive signals for those proteins were extremely suppressed in normal pancreatic tissues. All of the pancreatic cancer cell lines expressed IL-8 and IL-8 receptors at the RNA and protein levels. Receptor binding experiments using 125I-labeled IL-8 showed that PANC-1 cells had specific binding sites for IL-8. The cell proliferation assay demonstrated that IL-8 did not affect the growth of the three cell lines. However, treatment with IL-8 enhanced the invasiveness into Matrigel and increased the activity of matrix metalloproteinase (MMP)-2 in supernatants of the PANC-1 cells. These results demonstrate that IL-8 and IL-8 receptors are over expressed in pancreatic cancer, and suggest that IL-8 regulates MMP-2 activity and plays an important role in the invasiveness of human pancreatic cancer. PMID- 12632067 TI - Monocyte chemoattractant protein-1 expression correlates with macrophage infiltration and tumor vascularity in human gastric carcinomas. AB - The purpose of this study was to investigate the role of interactions between tumor cells and macrophages during angiogenesis in human gastric carcinomas. Macrophage infiltration into tumors and monocyte chemoattractant protein-1 (MCP 1) expression was assessed in 72 archival specimens of gastric carcinoma for comparison with tumor vascularity. The mRNA expression of MCP-1 was examined by RT-PCR in 6 gastric carcinoma cell lines and in fresh biopsy specimens from 18 patients. Immunolocalization of representative angiogenic factors, vascular endothelial growth factor (VEGF), and platelet-derived endothelial cell growth factor (PD-ECGF) was also done. MCP-1 expression in tumor cells increased with the depth of tumor invasion (Tis 9.5%, T1 19.4%, T2-4 60.0%), as did microvessel density and macrophage infiltration. Macrophage counts correlated with vessel counts, and both were significantly higher in MCP-1-positive than in negative tumors. Of the 6 gastric carcinoma cell lines, 2 constitutively expressed MCP-1 mRNA. In 6 (33.3%) of 18 biopsy samples, MCP-1 mRNA was expressed at higher levels in tumor tissues than in normal mucosa. VEGF protein was expressed by gastric carcinoma cells, whereas PD-ECGF protein was expressed mainly by stromal mononuclear cells. MCP-1 expression correlated significantly with VEGF but not PD ECGF expression in gastric carcinomas. These results suggest that MCP-1 produced by human gastric carcinoma cells plays a role in angiogenesis via macrophage recruitment and activation. PMID- 12632068 TI - Apolipoprotein E expression in localized prostate cancers. AB - Although prostate cancer has become the most common solid tumor diagnosed in men and the second leading cause of death due to malignancy, the progression of the disease is still somewhat unpredictable, until it becomes locally advanced or metastatic. The clinical problem to distinguish between dormant and progressive cancers is essential to patient care, as surgical and medical therapies have significant risk of morbidity and disability. Multiple gene expression in prostate cancer was surveyed by differential display cloning between the undifferentiated hormone refractory human prostate cancer cell line PC-3 (highly tumorigenic in nude mice) the hormone responsive LNCaP cell line (weakly tumorigenic in nude mice), and the poorly differentiated DU-145 cell line (moderately tumorigenic). ApoE mRNA was found to be highly expressed only in PC-3 cell-line. We further examined its protein expression by immunohistochemistry in 20 prostatectomy specimens and an association between Gleason score for each sample and ApoE expression was determined. ApoE expression correlates directly with the Gleason score in tissue sections, hormone independence as well as local and distant invasiveness. Prostatic intraepithelial neoplasias (PINs) adjacent to clinically manifested cancer were positive whereas more distant PINs were negative for ApoE. ApoE may represent a marker of more aggressive cells in human prostate cancers. PMID- 12632069 TI - Suppression of human prostate cancer cell growth by ciprofloxacin is associated with cell cycle arrest and apoptosis. AB - For hormone resistant prostate cancer (HRPC), chemotherapy is used but the mortality is 100% with a mean survival time of 7-8 months. Our previous studies have shown the chemotherapeutic effect of ciprofloxacin in bladder cancer. At doses 50-400 micro g/ml ciprofloxacin, the concentrations that are normally achieved at doses currently used for the treatment of anti-bacterial infections, inhibited bladder cancer cell growth and induced S/G2M arrest with modulation of key cell cycle regulatory genes and ultimately activated apoptotic processes. In this study, we investigated the effect of ciprofloxacin on androgen independent prostate carcinoma, PC3 cells and compared our results with non-tumorigenic prostate epithelial cells. The main advantage of this fluroquinolone antibiotic is its relative non-toxicity as compared to current chemotherapy, which is not very effective, for the treatment of advanced hormone resistant prostate cancer. PC3 cells as well as normal prostate epithelial cells (MLC8891) were treated with 25-400 micro g/ml ciprofloxacin, and cell counting was done during 3 days of treatment. The cell death was determined using DAPI staining of cell nuclei, 7AAD staining followed by flow cytometric analysis as well as by activation of caspase 3, a member of the ICE family of enzymes involved in the apoptotic cascade. The cell lysates were analyzed by immunoblotting techniques for the expression of key genes targeted by ciprofloxacin (p21WAF1, Bax and Bcl-2). Translocation of bax was visualized using a fluorescence staining procedure followed by laser confocal microscopic imaging. Treatment of prostate cancer cells with ciprofloxacin resulted in a dose- and time-dependent inhibition of cell growth (70-100% with 50 400 micro g/ml of the drug). There was a concomitant induction of cell cycle arrest at the S and G2/M phases of the cell cycle as well as induction of apoptosis. The CDK inhibitor p21WAF1 was down-regulated as early as 12 h following ciprofloxacin treatment (100-200 micro g/ml for 12-24 h). There was a significant increase in the Bax/Bcl-2 ratio with translocation of Bax, a pro apoptotic protein, to mitochondria with concomitant activation of caspase 3. These results suggest the potential usefulness of the fluroquinolone, ciprofloxacin as a chemotherapeutic agent for advanced prostate cancer. The fluroquinolone ciprofloxacin showed anti-proliferative and apoptosis inducing activity on prostate cancer cells but not on non-tumorigenic prostate epithelial cells. These effects of ciprofloxacin were mediated by cell cycle arrest at S G2/M phase of the cell cycle, Bax translocation to mitochondrial membrane and by increasing the Bax/Bcl-2 ratio in PC3 prostate cancer cells. Based on our in vitro results, further in-depth in vivo animal or human investigations are warranted. PMID- 12632070 TI - O(2) extraction is a key parameter determining the oxygenation status of malignant tumors and normal tissues. AB - The aim of this study was to assess pivotal factors determining the oxygenation status of malignant tumors, and, for comparison, of normal tissues. For this purpose, microvascular patterns, blood flow data, O2 consumption rates and O2 extractions were correlated to the median pO2 values of isografted rodent and xenografted human tumors, and of various normal tissues. No correlations were found between median pO2 values and vascular patterns, O2 consumption rate or blood flow rate. When O2 extraction, however, was considered (a comprehensive parameter which encompasses all of the above factors), a strong inverse correlation was found between this parameter and the median pO2 values, both for malignant tumors (r2=0.917) and normal tissues (r2=0.959). At similar O2 extraction values, the median pO2 values in the solid tumors were substantially lower than those in normal tissues, a finding which most probably reflects the chaotic microcirculation found in tumors, the loss of flow regulation which in normal tissues meets the metabolic demands of the cells, and in some cases, tumor associated anemia. PMID- 12632071 TI - Protein kinase Calpha regulates Ets1 transcriptional activity in invasive breast cancer cells. AB - We have previously shown that PKC inhibitors interfere with the Ets1/Smad3 dependent regulation of parathyroid hormone-related protein (PTHrP) P3 promoter activity by TGFbeta in invasive MDA-MB-231 breast cancer cells. By examining PKC expression in a variety of breast cancer cell lines, the protein level of PKCalpha was found to be much higher in Ets1-expressing MDA-MB-231 and MDA-MB-435 breast cancer cells than in Ets1-deficient MCF-7 and SK-BR3 cells. No correlation of Ets1 expression with the expression of other PKC subtypes (PKCbeta1, PKCbeta2, PKCdelta or PKCepsilon) could be observed. In contrast to MDA-MB-231 cells, PKCalpha-deficient MCF-7 cells do not support Ets1-induced activation of the PTHrP P3 promoter suggesting that PKCalpha may be important for Ets1 activity. A constitutively active form of PKCalpha was found to potentiate the P3 promoter activation by Ets1 alone and in synergy with Smad3. PKCalpha, but not PKCepsilon, also induced phosphorylation of the Ets1 protein. Both PKCalpha effects on Ets1 depended on the exon VII domain of Ets1. Using verapamil and ionomycin, we could show that PKCalpha induces Ets1 phosphorylation independent of calcium mobilization. Collectively, our data suggest that PKCalpha may regulate Ets1 activity in invasive breast cancer cells. PMID- 12632072 TI - Fluorescent-based BAT-26 analysis for distinct screening of microsatellite instability in colorectal cancers. AB - For easy screening of genetic instability in colorectal cancers, we tried BAT-26 and BAT-25 mononucleotide repeats using fluorescent analysis and evaluated their usefulness and problems compared with other markers: D5S346, D17S250, D2S123, and D2S391, D4S392 (located near BAT-26 and BAT-25 respectively). The high-frequency of MSI (MSI-H) tumours, defined as tumours having instability in more than two markers, were detected in 8/146 (5.5%). These MSI-H cases were younger ages at diagnosis, and showed significantly higher incidences of right side location, and poorly differentiated histology than other cases (p<0.05). Four cases (2.7%) showed a 1 bp size shift in BAT-26 and 2 of them showed loss of heterozygosity (LOH) at D2S391 near BAT-26 locus. Among 9 cases with a 1 bp size shift in BAT 25, 6 cases showed LOH at D4S392 near the BAT-25 locus (p=0.035). In all 4 cases, non-cancerous DNA had two analogous peaks of BAT-26, indicating the heterozygosity of BAT-26 in constitutional DNA. This phenomenon was also detected in the peaks of BAT-25 in some cases, in whose constitutional DNAs, 1 bp size shift was also detectable in three other markers. To elucidate the reasons for the alterations of the 1 bp size shift of peak of these markers, we examined by microsatellite analysis mixed samples of tumour DNA with complete loss of the one allele at the 1p loci and each constitutional DNA sample of neuroblastoma patients. One base shift of the peak signal of the microsatellite marker was clearly obtained in proportion to the ratio of cancerous DNA and constitutional DNA. Fluorescent-based analysis of BAT-26 or BAT-25 was easy and useful for detection of MSI-H in colorectal cancers without analyzing non-cancerous DNA. A 1 bp size shift in BAT-26 or BAT-25 was considered to be affected by LOH at these loci. Thus, it is important to distinguish MSI from LOH to evaluated MSI using these markers. PMID- 12632073 TI - High expression of the novel endothelin-converting enzyme genes, Nbla03145/ECEL1alpha and beta, is associated with favorable prognosis in human neuroblastomas. AB - The clinicobiological feature of neuroblastoma is enigmatic because spontaneous regression often occurs in early stages of tumors of the patients under 1 year of age, while rapid growth usually occurs in the tumors of the patients over 1 year of age. Such difference in the clinical behavior may be caused by the difference in the pattern of gene expression among the subsets of neuroblastoma. To understand the molecular basis of neuroblastoma biology, we decided to identify the novel genes expressed differentially between favorable and unfavorable neuroblastomas. The oligo-capping cDNA libraries were constructed from different subsets of neuroblastomas. After random selection and DNA sequencing, the differentially expressed genes between favorable and unfavorable neuroblastomas were screened by reverse transcriptase-PCR. The clinical significance of gene expression was evaluated based on the results of Northern blot analysis. We have identified a novel gene Nbla03145 (alpha), also cloned and termed by another group as ECEL1, which encodes a new member of putative zinc-binding metalloendopeptidase (endothelin-converting enzyme) with unknown substrate. We also cloned a COOH-terminally truncated Nbla03145/ECEL1beta which is expressed only in thymus. In primary NBLs, the alpha isoform is more preferentially expressed than the beta isoform. High levels of Nbla03145/ECEL1 expression were significantly correlated with a younger age (p=0.0005), lower stages (p=0.0019), high level of TrkA expression (p 8 mm, cholestasis, and acute biliary pancreatitis. RESULTS: ERC for suspected choledocholithiasis was performed in 310 patients. The rate of successful cannulation for ERC was 96.8%. Duct stones were found in 86%. Twelve patients had impacted duct stones cleared at open common duct exploration. The failure rate of ERC was 4.5%, and the rate of unnecessary ERC was 13.5%. LC was performed in 298 of 310 patients. Morbidity rates were 2.2% and 1.5% after ESE and LC, respectively. CONCLUSION: A total of 95.5% of patients with symptomatic gallbladder and ductal stones could be successfully managed by ERC prior to LC with a low morbidity rate. PMID- 12632132 TI - Helium pneumoperitoneum reduces tumor recurrence after curative laparoscopic liver resection in rats in a tumor-bearing small animal model. AB - BACKGROUND: After exposure of neoplastic tissue to helium, a significant reduction of tumor growth has been detected in experimental studies, both in vitro and in vivo. This tumor-suppressive effect of helium is controversly discussed in the literature. It was therefore the aim of this study to investigate the influence of pneumoperitoneum with CO2, room air, or helium in a tumor-bearing small animal model comparing laparoscopic partial hepatic resection for hepatocellular carcinoma with conventional open partial hepatectomy. METHODS: One-hundred forty-eight male American Cancer Institute rats underwent partial hepatectomy for curative resection of previously induced hepatocellular carcinoma (Morris hepatoma 3924A). Resection was performed either in open laparotomy (n = 30) or laparoscopically under the employment of CO2 (n = 30), room air (n = 30), or helium (n = 30) for the pneumoperitoneum. Twenty-eight animals served as controls receiving anesthesia but no tumor resection. All animals were sacrificed on postoperative days 21, 35, or 56 for autopsy and evaluation of possible tumor recurrence and metastasis. RESULTS: Significant reduction of postoperative tumor recurrence and metastasis was observed in the group of animals receiving laparoscopic tumor resection under helium insufflation compared to open surgery or laparoscopic resection with air pneumoperitoneum. CONCLUSIONS: The results of this study suggest a suppressive effect of helium pneumoperitoneum on postoperative tumor growth and metastatic spread. Furthermore, tumor exposure to room air appears to have a stimulative influence on tumor recurrence and metastasis compared to a pneumoperitoneum established with CO2. PMID- 12632133 TI - Oncological quality and preliminary long-term results in laparoscopic colorectal surgery. AB - BACKGROUND: Our aim here was interpret data on the perioperative course, oncological quality, and preliminary long-term results of laparoscopic colorectal surgery carried out with a curative intent. METHODS: The data were collected within the framework of a prospective multicenter observational study that has been ongoing since 1 Aug 1995 and includes 46 hospitals. Of a total of 3133 patients, 826 (26.4%) underwent a curative resection for colorectal carcinoma. RESULTS: The average age of the patients was 67.9 years; the sex distribution was almost 1:1. UICC staging of tumors (stages I, II, and III) showed the following figures: 301/36.4%, 265/32.1%, and 260/31.5%. In the majority of cases, an oncologically radical resection with high transection of the supplying vessels was performed. Intraoperative seeding of tumor cells was reported in 1.8% of the patients. In eight cases, the seeding was due to spontaneous rupture of the tumor. A mean of 13.5 lymph nodes in the resected specimen were investigated histopathologically (10.9 lymph nodes in stage I, 15 each in stages II and III). Depending on the individual hospital, we found a remarkable variation in the number of lymph nodes investigated. With a mean follow-up period of 2.1 years, Kaplan-Meier survival function showed acceptable results, both for rectal and colonic carcinoma, in comparison with conventional colorectal surgery. A stage related consideration of the survival data yielded similar results. CONCLUSION: All in all, the results show that a laparoscopic colorectal procedure can meet oncological radicality criteria, even though certain reservations-in particular, in the case of procedures done with a curative intent-have not been completely eliminated. PMID- 12632134 TI - National trends in utilization and outcomes of antireflux surgery. AB - BACKGROUND: Studies examining the outcomes of surgery for gastroesophageal reflux disease (GERD) have consisted primarily of case series. We sought to assess trends in both utilization and outcomes of antireflux surgery from a national perspective. METHODS: Using ICD-9 codes, we identified all antireflux procedures (N = 24,208) performed on adults from 1990 to 1997 in hospitals participating in the Nationwide Inpatient Sample, the largest all-payer inpatient care database in the United States. Using sampling weights and U.S. Census data, we then calculated the national population-based rate of antireflux surgery for each year and examined secular trends in utilization, in-hospital mortality, splenectomy (a technical complication), and length of hospital stay. Using a coding algorithm, we also assessed trends in the proportion of procedures performed via the laparoscopic, open abdominal, and thoracic approaches. RESULTS: From 1990 to 1997, the population-based annual rate of antireflux surgery increased from 4.4 to 12.0 per 100,000 adults. A substantial increase in utilization was observed from 1993 to 1995, but annual rates before and after this period were relatively stable. Between 1990 and 1997, in-hospital surgical mortality decreased from 1.2% to 0.5% (p = 0.002), splenectomy rates decreased from 3.9% to 1.5% (p <0.001), and median length of stay decreased from 7 to 2 days (p <0.01). The proportion of antireflux procedures performed laparoscopically increased from 0.5% to 64% (p <0.001), and the proportion of procedures performed using a thoracic approach decreased from 12% to 1% (p <0.001). CONCLUSIONS: With the dissemination of the laparoscopic approach, the population-based rate of antireflux surgery has more than doubled. At the same time, operative mortality and splenectomy risks have diminished. PMID- 12632136 TI - Hand-assisted laparoscopic splenectomy for giant spleens. AB - BACKGROUND: Laparoscopic splenectomy for massive splenomegaly is technically difficult, and the morcellated splenic tissue may be inadequate for histologic study. A hand-assisted technique may provide a technical advantage and allow removal of larger pieces of spleen. METHODS: Patients who underwent hand-assisted laparoscopic splenectomy for massive splenomegaly were reviewed. Demographic information, operative data, and outcomes data were tabulated. RESULTS: Sixteen patients met these criteria. Mean age was 56 years (range, 35-78 years). Operating time averaged 240 min (range, 165-360 min), and median blood loss was 425 cc (range, 100-1800 cc). There were no conversions to an open procedure. Mean weight of extracted spleens was 2008 g (range, 543-4090 g). Postoperative length of stay averaged 3.3 days (range, 2-7 days). There was one postoperative complication (6.25%) and no mortality. CONCLUSIONS: Hand-assisted laparoscopic splenectomy for massive splenomegaly is feasible and safe while preserving the recovery benefits of minimal access surgery. It provides an adequate specimen for histologic study. PMID- 12632135 TI - Digital stereotactic biopsies for nonpalpable breast lesion. AB - BACKGROUND: Percutaneous biopsy (BP) is a valid alternative to open surgical biopsy. The aim of our study was to evaluate the results and diagnostic value of vacuum-assisted core biopsy (VACB; Mammotome) and advanced breast biopsy instrumentation (ABBI). METHODS: From June 1999 to December 2001, 360 BPs were performed: all patients had dubious mammography lesions not confirmed by ultrasonography. Indications were as follows 264 (73.3%) microcalcifications, 64 (17.8%) nodular opacities, and 32 (8.8%) parenchymal distortions. RESULTS: All BPs were performed with a digital stereotactic table with a vacuum suction aspiration system for VACB and a cutting cannula for ABBI. All BPs were correctly performed. Seventy-one (19.7%) lesions were malignant, whereas 258 (71.6%) were benign: 31 (8.6%) of the lesions removed with VACB were atypical ductal hyperplasia. CONCLUSIONS: BP is a valid method for the diagnosis of nonpalpable breast lesions. In our experience, VACB is the method of choice because it is easy to perform and has high adaptability. PMID- 12632137 TI - Intrapleural analgesia following thoracoscopic sympathectomy for palmar hyperhidrosis: a prospective, randomized trial. AB - BACKGROUND: Reports on intrapleural analgesia (IPA) are conflicting. The current study assessed the effect of a single-dose thoracoscopic bilateral intrapleural anesthetic administration on the immediate postoperative recovery room and 24-h pain control. METHODS: Fifty patients with primary palmar hyperhidrosis were randomly classified into two groups to receive either 20 ml of 0.5% bupivacaine and 5 mg/ml epinephrine or 0.9% NaCl in each thoracic cavity at the end of thoracoscopic T2-T3 sympathectomy. The degree of early postoperative pain was estimated by visual analog scale (VAS). The 24-h parenteral opioid analgesic requirement was recorded. RESULTS: The immediate postoperative VAS score (1.46 +/ 0.41 vs 2.0 +/- 0.61, p = 0.03), opioid consumption (0.42 +/- 0.36 vs 0.65 +/- 0.28, p = 0.0133), and 24-h opioid consumption (1.02 +/- 0.80 vs 1.48 +/- 0.84, p = 0.05) were significantly reduced following IPA compared to those of the control group. CONCLUSION: IPA is a simple and effective means for postoperative pain control following thoracoscopic upper dorsal sympathectomy. PMID- 12632138 TI - A lay person versus a trained endoscopist: can the preop endoscopy simulator detect a difference? AB - The purpose of this study was to establish construct validation of a flexible sigmoidoscopy simulator by comparing training-level grouped subjects. These included clerical staff (n = 10), residents (n = 19), and experts (n = 5). Each participant performed 3 scopes. The ANOVA group-based results for trainer measured variables are shown in Table 1. These results demonstrate that the flexible sigmoidoscopy simulator distinguished the trained from the untrained and the resident from the expert. Although there was no statistically significant differences between the senior residents and the experts, the expert commonly outperformed the residents. Establishing the transferability of simulator training to real life is next. If the transfer of skill can be established, it may give rise to a new skills training approach. PMID- 12632139 TI - Staged and complete laparoscopic management of cholelithiasis in a patient with gallstone ileus and bile duct calculi. AB - BACKGROUND: Gallstone ileus is an uncommon cause of small bowel obstruction, and its incidence peaks in elderly women. Although enterolithotomy has been accomplished laparoscopically, often using a laparoscopically assisted approach, controversy persists as to the indication, timing, and surgical approach to a cholecystectomy with closure of the cholecystoduodenal fistula. METHODS: We present the case of a 63-year-old woman with symptomatic cholecystolithiasis who presented with acute gallstone ileus and underwent an emergency laparoscopic enterolithotomy. Hypotonic duodenography during the follow-up period demonstrated a cholecystoduodenal fistula and previously unsuspected stones in the bile duct. The patient underwent an elective laparoscopic cholecystectomy with repair of the fistula, a concomitant bile duct exploration, choledocholithotomy, and primary bile duct closure. RESULTS: The patient enjoyed an uneventful recovery, and was discharged home on postoperative day 5 after her initial emergency surgery. Her recovery after the subsequent elective surgery was more expeditious, with a discharge from hospital on postoperative day 2 and a return to office employment 2 weeks later. CONCLUSION: In the good-risk patient, staged laparoscopic management of gallstone ileus and the associated cholecystoduodenal fistula is feasible and appears to be safe. In such patients, imaging of the biliary tree is essential to detect silent choledocholithiasis, which also may be managed concomitantly and safely by the laparoscopic approach. PMID- 12632142 TI - Growth of whaleworm (Anisakis simplex, Nematodes, Ascaridoidea, Anisakidae) third stage larvae in paratenic fish hosts. AB - The growth of Anisakis simplex L3 larvae in the three paratenic hosts saithe ( Pollachius virens), cod (Gadus morhua) and redfish (Sebastes marinus) was studied based on material collected off the island of Vega on the west coast of Norway over a period of 1 year. In all three fish species, a positive correlation between the length of larvae and the age of the fish was shown. The number of large larvae, here defined as L3>28 mm, increases with increasing age of the host. These findings clearly indicate an accumulation of large L3 larvae and larval growth in the fish host. The larvae were additionally grouped according to the tissue--muscle, liver or other viscera--they were located in. When the average lengths of the three groups of larvae were compared, the gadoids saithe and cod had somewhat higher values for L3 larvae found in the liver. Redfish, on the other hand, which store fat to a relatively higher degree in the viscera, had the highest average length of larvae in this tissue. The lowest average values in all three species were found in the musculature. These trends suggest that the growth of A. simplex L3 larvae is positively correlated with the fat content of the tissue in which it is encapsulated. Of the three host species included in this study, saithe seemed to be the best suited for A. simplex L3, as the growth rate and average length of the parasite was greatest in this species. The growth and average length of L3 was least pronounced in redfish while cod held an intermediate position. PMID- 12632143 TI - Cyst burden in the brains of Wistar rats fed Toxoplasma oocysts. AB - Six strains of Toxoplasma oocysts were used to infect groups of 4-24 Wistar rats, with each rat being fed 10(1)-10(4) oocysts from a single strain. After 2 months, the rats were killed, their brains screened for Toxoplasma cysts and then bioassayed in mice if negative. Toxoplasma was either observed in the form of brain cysts or was recovered using the bioassay, from 113 out of 138 (82%) rat brains. As few as ten oocysts were capable of initiating a brain infection that lasted for at least 2 months in eight of the nine rats inoculated. However, judging from bioassay 10(2)-10(4) oocysts did not give rise to progressively higher rates of infection. Brain cysts were seen in only 68 of 138 rats (49%). The number of Toxoplasma cysts formed in the brains of rats was generally in the order of tens to hundreds. The frequency of infection in the brains with Toxoplasma and the number of brain cysts formed appeared to be influenced by the individual resistance of the rats as well as by the doses of oocysts and the Toxoplasma strains used. The information gathered is considered to be a basis for a rat model of immunity against acquired toxoplasmosis. PMID- 12632144 TI - Ultrastructure of the spermatozoon of the proteocephalidean cestode Proteocephalus torulosus (Batsch, 1786). AB - The fine structure of the mature spermatozoon of the proteocephalidean tapeworm Proteocephalus torulosus, a parasite of cyprinid fishes, was studied by transmission electron microscopy for the first time. The mature spermatozoon of P. torulosus is filiform and tapered at both extremities. It contains two axonemes of the 9+"1" type and of unequal length. The anterior extremity of the spermatozoon bears a helicoidal crested body approximately 100 nm thick. Cortical microtubules of two types lie parallel to the spermatozoon axis. The nucleus is a fine cord of condensed chromatin. The slightly electron-dense cytoplasm contains electron-dense granules in regions II and III of the spermatozoon. The anterior and posterior extremities of the spermatozoon contain a single axoneme. The mature spermatozoon of P. torulosus differs from that of other Proteocephalusspecies in the presence of a single crested body and in the morphology of its posterior end. PMID- 12632145 TI - Fetal Toxoplasma infection after oocyst inoculation of pregnant rats. AB - Six groups totalling 53 Wistar rats were fed 10(4)oocysts from one of six different Toxoplasma strains at 15 days of pregnancy. The overall transplacental transmission rate was 51%. This varied between 10% and 80%, dependent on the strain used. The strains of Toxoplasma which are more pathogenic for mice were transmitted transplacentally more frequently than the strains of intermediate or low pathogenicity. There were no statistically significant differences in the rate of congenital transmission of Toxoplasmain rats fed oocysts (present work) or cysts (previous work). PMID- 12632146 TI - Comparison of three antigen detection tests for diagnosis and follow-up of falciparum malaria in travellers returning to Berlin, Germany. AB - We determined the sensitivity and specificity of three rapid immunochromatographic malarial antigen detection test systems (RDTs) for the detection of Plasmodium falciparumand assessed the quality of follow-up results. ParaSight-F and ICT Malaria detect histidine-rich protein-2 (HRP-2), whereas OptiMal detects plasmodial lactate dehydrogenase (pLDH). ParaSight-F performed with 95.1% sensitivity and 97.1% specificity (554 patients tested of whom 144 had falciparum malaria). ICT Malaria performed with 95.7% sensitivity and 99.2% specificity (718 patients tested of whom 184 had falciparum malaria). OptiMal performed with 76.2% sensitivity and 99.7% specificity (539 patients tested of whom 130 had falciparum malaria). In follow-up investigations, HRP-2 did not appear to be a useful antigen due to its long half-life, whereas pLDH offers a reasonable correlation with the presence of viable parasites in those cases initially detected. We therefore conclude that a combination of both antigens might be the best option for creating a reliable RDT for the diagnosis of falciparum malaria. PMID- 12632147 TI - Comparative karyological analysis of three species of Bothriocephalus Rudolphi 1808 (Cestoda: Pseudophyllidea). AB - The diploid complements of mitotic metaphase plates of three species of Botriocephaluswere studied using conventional Giemsa staining and karyometric analysis. Botriocephalus claviceps and Botriocephalus gregarius showed a karyotype with 2n=14 chromosomes. All chromosomes of B. claviceps were biarmed, metacentric or meta-submetacentric. The two first pairs of homologues were markedly larger than the remaining elements and represented 48.62% of the total chromosome length. The karyotype of B. gregarius had similar metric values for the chromosomes, but clearly differed in the chromosome morphology of pairs 5, 6 and 7, which had terminally located centromeres. The diploid number 2n=12 was found in Botriocephalus scorpii. The karyotype consists of five pairs of metacentric and one pair of submeta-metacentric chromosomes. The possible pathways of karyotype evolution within Bothriocephalus spp. and their phylogenetic relations with the other karyologically studied groups in the order Pseudophyllidea are discussed. PMID- 12632149 TI - Design and development of an immunosensor for the detection of malaria in field conditions. AB - Eradication of malaria in Southeast Asian countries is still a distant goal, due to the absence of a simple, rapid and inexpensive diagnostic technique. Here, an immunosensor for the photometric detection of malaria, the malaria-detecting immunosensor (MDI), is developed to detect Plasmodium falciparum malarial antibodies in human blood. The method uses the principle of laser light scattering by latex bead agglutinates in media monitored by a light-detecting device. Agglutination is induced by mixing antigen-coated latex beads with serum antibodies. Immunoreactions are measured in terms of the Tyndall effect in the transmitted beam detected by photodiodes. MDI sensitivity and specificity are compared with the results of enzyme-linked immunoabsorbent assay and laser light scattering immunoassay techniques, which show that it is a good and sensitive monitoring device. PMID- 12632148 TI - Intestinal peptide transporter PepT1 is over-expressed during acute cryptosporidiosis in suckling rats as a result of both malnutrition and experimental parasite infection. AB - Cryptosporidium parvuminfection induces amino acid malnutrition leading to growth retardation in children. Owing to the nutritional efficiency of peptides compared to free amino acids and the resistance of the di-tripeptide transporter PepT1 to mucosal injury, we analyzed the intestinal expression of PepT1 during experimental acute cryptosporidiosis in suckling rats from day 4 to day 50. PepT1 mRNA levels were increased at the peak of infection (day 10) all along the small intestine and normalized after spontaneous clearance of the parasite (day 21). Immunolocalization of PepT1 showed that its expression was maintained in the brush border membrane of enterocytes in infected rats from day 4 to day 50 all along the small intestine. Our results suggest a transcriptional up-regulation during acute cryptosporidiosis in response to both C. parvum-induced malnutrition and parasite implantation. As no treatment is available, a semi-elemental diet should be considered part of the treatment of cryptosporidiosis. PMID- 12632150 TI - Gene structure of Taenia solium paramyosin. AB - Paramyosin is a muscle protein that probably plays a role in the survival of the larval stage of Taenia solium during its prolonged host-parasite relationship. Here we describe the structure of the gene coding for the paramyosin of T. solium. The characterization of two clones obtained from a genomic library showed that the complete gene of paramyosin contains 13 introns delimited by conventional eukaryotic splice signals. Comparison with the paramyosin genes of Drosophila melanogaster and Caenorhabditis elegans showed a lack of conservation of the exon/intron organization in contrast to other muscle genes. No evidence of alternative splicing sites were found, excluding the possibility that T. solium expresses a mini-paramyosin like D. melanogaster. PMID- 12632152 TI - Immunohistochemical/immunogold detection and distribution of the endosymbiont Wolbachia of Dirofilaria immitis and Brugia pahangi using a polyclonal antiserum raised against WSP (Wolbachia surface protein). AB - Intracellular bacteria in filarial nematodes were described as early as the 1970s, yet it was only with the work on Dirofilaria immitis, the agent of canine and feline heartworm disease, that these microorganisms were identified as belonging to Wolbachia, a genus known for encompassing bacteria infecting insects and other arthropods. The implications for the presence of intracellular bacteria in filarial nematodes is now the subject of intense research, particularly regarding their role in the immunology and pathogenesis of disease in infected humans and animals and as a possible target for therapy. Here, the authors report results on the immunohistochemical and immunogold staining of Wolbachia in D. immitis and Brugia pahangi using polyclonal antibodies raised against the recombinant Wolbachia surface protein (WSP). The bacteria were present in the lateral hypodermal chords of both male and female worms and in the reproductive tract of adult females (oocytes, morulae, microfilariae). In D. immitis and B. pahangi from animals treated with tetracycline, positive staining was observed in the lateral chords of adult males and females, but was absent from the oocytes and morulae. These results indicate that Wolbachia endosymbionts can be identified immunohistochemically with anti-WSP polyclonal antibodies, that their distribution matches that already described for Wolbachia of other filarial worms, and that antibiotic treatment may impede the vertical transmission of these bacteria. Unequivocal detection of Wolbachia is essential for the study of this symbiont, in particular to monitor the effects of antibiotic treatment on worms. The use of a specific marker for bacteria in their nematode hosts represents an extremely useful tool in evaluating the pathogenic role and the effect of antibiotic treatment on these potential targets in the control of filarial disease. PMID- 12632151 TI - Ultrastructural localization of Ani s 1, a major allergen from the fish parasite Anisakis simplex. AB - No data about the nature or function of the first major Anisakis simplex allergen, named Ani s 1, is available. The aim of this study was to investigate the ultrastructural localization of this protein, to obtain further information about it. Ani s 1 was detected in secretory granules of the excretory gland and occasionally lining the main excretory canal. Our results suggest that Ani s 1 could be a secretory product and could have an enzymatic function related to mechanisms of infection. PMID- 12632153 TI - Population dynamics of sealworm, Pseudoterranova decipienssensu lato, in sculpins, Myoxocephalus scorpius, from two areas in Norway between 1990 and 1996. AB - The abundance of sealworms (Pseudoterranova decipienss.l.) in sculpins (Myoxocephalus scorpius) was studied at Torbjornskjaer, southeast Norway, and Vega, northern Norway, in samples from 1990 to 1996. The harbour seal ( Phoca vitulina) population at Torbjornskjaer declined by 70% during the phocine distemper virus epizootic in 1988, and has increased during the 1990s. The abundance of sealworms in sculpins was relatively stable from 1992 to 1996 and any effect of the reduction in the number of harbour seals on the abundance of sealworms has been short-term. When correcting for size differences, young sculpins (2-4 years) from Torbjornskjaer had higher abundances than those from Vega, while in old sculpins (5-7 years) the abundances seemed similar and stabilised. This happened at a younger age among the more heavily infected sculpins at Torbjornskjaer than at Vega, and we interpret this as density dependence in the fish host. PMID- 12632154 TI - Molecular characterization of Blastocystis isolates from birds by PCR with diagnostic primers and restriction fragment length polymorphism analysis of the small subunit ribosomal RNA gene. AB - Seven Blastocystis isolates from birds were genotyped by polymerase chain reaction (PCR) using diagnostic primers, and the homology among isolates was then confirmed by restriction fragment length polymorphism (RFLP) analysis of the small subunit ribosomal RNA gene. A zoonotic genotype, subtype 4, was detected in three isolates by PCR, but the RFLP profiles of these isolates differed from those of the subtype 4 reported previously from a human. None of the other four isolates were amplified with any subtype-specific primer sets, and the profiles of these isolates were different from any ribodemes of Blastocystis hominis reported previously. In the present study, the seven isolates from birds were classified into three genetically distinct genotypes. Therefore, birds appear to be infected with multiple genotypes. It is unclear whether the isolates examined in the present study are zoonotic or not. Therefore, it is important to determine the phylogenetic relationships between unknown genotypes from birds and the multiple genotypes of B. hominis. PMID- 12632155 TI - Further studies on the structural analysis of the cuticle of Litomosoides chagasfilhoi (Nematoda: Filarioidea). AB - In order to obtain further information on the structural organization of the cuticle of nematodes, this structure was isolated from adult forms of the filariid Litomosoides chagasfilhoi. The purity of the fraction was determined by light and transmission electron microscopy, deep-etching, high resolution scanning electron microscopy, atomic force microscopy, immunocytochemistry, gel electrophoresis (SDS-PAGE) and Western blot. The epicuticle presented a rugous surface with parallel rows and several globular particles that could be involved in the absorption of nutrients and secretion of products. Analysis by SDS-PAGE of purified cuticles revealed five major polypeptides corresponding to 151, 41, 28, 13 and 11 kDa. A polyclonal antibody against a synthetic 18 amino-acid peptide that corresponds to the sequence of domain E of the Haemonchus contortus3A3 collagen gene recognized several protein bands on the Western blot of purified cuticle, and labeled all cuticular layers, as shown by immunocytochemistry. PMID- 12632157 TI - Liver enzyme activity and histological changes in the liver of silver foxes (Vulpes vulpes fulva) experimentally infected with opisthorchiid liver flukes. A contribution to the pathogenesis of opisthorchiidosis. AB - Blood samples from silver foxes experimentally infected with Opisthorchis felineus and Metorchis bilis, respectively, were examined for the activity of liver enzymes. The average activities of aspartate aminotransferase (AST), glutamate dehydrogenase (GLDH), alkaline phosphatase and alanine aminotransferase in uninfected control animals were 20, 1.8, 57 and 44 units/l, respectively. The liver enzymes in infected foxes reacted differently, depending on dose, species of flukes and individual peculiarities. The highest individual deviation of infected from control animals was registered in the case of GLDH, reaching increases of up to 200-fold. In contrast, AST showed the lowest deviation from control values (less than 10-fold). By the end of the study period, enzyme activities had declined. The prepatent periods for M. bilis and O. felineus in foxes were 2 weeks and 4 weeks, respectively. High egg per gram values were established at the beginning of the patent period. At necropsy, chronic inflammatory reactions were found in the bile ducts and in the wall of the gall bladder. The number of flukes at the end of the study was low. PMID- 12632156 TI - The initial immune response during experimental cysticercosis is of the mixed Th1/Th2 type. AB - The immunological events that occur during the initial stages of experimental cysticercosis are not known. The studies presented here examined the cytokines produced by peritoneal exudate cells (PECs), splenocytes and mesenteric lymph node (MLN) cells during the first week of infection with larval Taenia crassiceps in BALB/cJ mice. Proliferation assays determined that the earliest time when antigen-specific responses could be measured was 5 days post-infection. Concanavalin A (ConA) stimulation of host cells elicited an initial burst of IL-4 production at 24 h of infection and ConA-stimulated Th2-type cytokine production is predominant by 7 days post-infection. Thus, there are responses at day 1 of infection that seem to promote a Th2-type response. Stimulation of MLN cells, splenocytes and PECs with larval antigens supported previous reports of mixed Th1/Th2-type cytokine production with increases in interleukin (IL)-4, IL-10 and interferon (IFN)-gamma. Ex vivo IFN-gamma production by PECs from infected mice was increased at 3, 5 and 7 days post-infection, whereas at these times reduced ex vivo IL-10 production was observed. This ex vivo IFN-gamma response preceded an increasing IL-10 production by PECs between 3 and 7 days post-infection in parasite-specific and ConA-induced proliferation assays. Thus, infection with larval T. crassiceps results in an initial response mediated by IFN-gamma that is quickly followed by an increase in IL-10 production and subsequent reduction in the amount of IFN-gamma being produced. PMID- 12632159 TI - Identification of genotypes of Cryptosporidium parvum isolates from ferrets in Japan. AB - Recent molecular studies show that Cryptosporidium parvum is composed of at least eight genotypes including zoonotic genotypes. Therefore, it is important to screen the genotypes of the isolates harbored in animals for the control of human cryptosporidiosis. The ferret is a popular pet, but also a reservoir of Cryptosporidium. Since the infectivity of zoonotic genotypes in ferrets remains unclear, there is a possibility these hosts harbor zoonotic genotypes. In the present study, we examined the genotypes of C. parvum isolates from ferrets in Japan using polymerase chain reaction direct sequencing. The sequences of the isolates examined clustered with the ferret-adapted genotype (ferret genotype). Our study suggests that ferrets harbor the ferret genotype which is conserved across geographical areas. Further study is required to determine whether ferrets are a significant reservoir for human cryptosporidiosis. PMID- 12632158 TI - Is the genus Digramma synonymous to the genus Ligula (Cestoda: Pseudophyllidea)? Evidence from ITS and 5' end 28S rDNA sequences. AB - The genus Digramma (Cestoda: Pseudophyllidea) described by Cholodkovsky in 1915 differs from the genus Ligula only by the number of the reproductive organs per proglottis. However, the occurrence of transitional forms in Digramma raises much confusion concerning its generic validity. In the present study, cestodes previously designated as Digramma and Ligula were collected from lakes in the lower and middle reaches of the Yangtze River, and also from Qinghai Lake on Qingzang plateau, China. The entire internal transcribed spacer of the ribosomal DNA (ITS rDNA) and 5' end of 28S rDNA were compared between the Digramma and Ligula specimens. The low level of nucleotide variation between the two genera may imply that cestodes in the genus Digramma are paraphyletic to the Ligula genus, and Digramma is a synonym of Ligula. However, whether previously identified Digramma cestodes represent different species in the genus Ligula requires further investigation. PMID- 12632160 TI - Metazoan parasites and food composition of juvenile Etmopterus spinax (L., 1758) (Dalatiidae, Squaliformes) from the Norwegian Deep. AB - A total of 37 juvenile Etmopterus spinax from the Norwegian Deep were examined for stomach contents and metazoan ecto- and endoparasites. These squaliform elasmobranchs were caught by benthopelagic net in May 2001 at a depth of 194-214 m. The euphausiid Meganyctiphanes norvegica and the teleost Maurolicus muelleri were the principal prey organisms. With increasing total length of E. spinax, the frequency of Meganyctiphanes norvegica prey decreased and that of M. muelleri increased. Seven different metazoan parasite species were found: adult Monogenea (2), larval and adult Cestoda (3), and larval Nematoda (2). The predominant parasites were an unidentified monocotylid monogenean and the cestode Aporhynchus norvegicus, with respective prevalences of infestation of 83.8% and 81.1%. The sites of infestation were the gills ( Squalonchocotyle spinacis, Monogenea), nasal cavities (Monocotylidae indet.), body cavity ( Lacistorhynchus tenuis, Cestoda), stomach wall and organs of the body cavity ( Anisakis simplex, Nematoda), and stomach and spiral valve ( A. norvegicus and Pseudophyllidea indet., Cestoda; Hysterothylacium aduncum, Nematoda). No other metazoan parasite taxa were found, and the musculature was free of parasites. Five new host and three new locality records were established. M. muelleri seems to be an important intermediate host for the endoparasitic nematodes which were found, with E. spinax serving as a paratenic host. E. spinax also serves as an intermediate host for the trypanorhynch cestode L. tenuis, and as the definitive host for the two monogeneans and the trypanorhynch A. norvegicus. The latter uses Meganyctiphanes norvegica as the second intermediate host in the Norwegian Deep. The relationship between the feeding ecology, habitat, and vagility of E. spinax and the resulting parasite fauna is discussed. PMID- 12632161 TI - Individual cases of autochthonous malaria in Evros Province, northern Greece: entomological aspects. AB - Prompted by four autochthonous cases of malaria in 1994 and 1995 in Evros Province, northern Greece, we conducted an entomological study between 1997 and 1999 in Nipsa and Chandras, rural locations where two of the four cases had occurred, and in Feres where two additional autochthonous malaria cases had been diagnosed in 1998. In Nipsa and Chandras, we identified 29 Anopheles breeding sites and characterized them by physicochemical parameters. Larvae were collected both at these sites and in a brackish water breeding site near Feres in the Evros River delta. Adults were caught in sheds at all three locations. Morphology was used to classify larvae and adults as A. superpictusor as species belonging to the A. claviger or A. maculipennis species complexes. The latter were further identified by PCR as being A. maculipennis s.s., A. melanoon and A. sacharovi. Of the A. maculipennis complex larvae collected inland, approximately 94% were A. maculipennis s.s. and 6% A. melanoon, whereas all larvae collected in the coastal region were A. sacharovi. In contrast, the A. maculipennis adults were A. maculipennis s.s. and A. melanoon (both 47%), and A. sacharovi (6%). In the coastal region, no A. maculipennis s.s. adults were caught. The ratio of A. melanoon adults collected to A. sacharovi was about 3:1. As shown by a bloodmeal ELISA, only 5 of 266 fed females (1.9%) had ingested human blood, whereas 232 (87%) had fed on goats. Of the mosquitoes containing human blood, two were A. melanoon, one A. sacharovi and one A. maculipennis s.s. One human blood specimen could no longer be assigned to a particular mosquito. PMID- 12632162 TI - Expression and characterization of recombinant pyruvate kinase from Toxoplasma gondii tachyzoites. AB - We have cloned a cDNA encoding Toxoplasma gondii pyruvate kinase and obtained the full-length recombinant enzyme with a calculated molecular mass of 57.5 kDa. The predicted amino acid sequence of T. gondii pyruvate kinase exhibited a highest identity (63%) to that of Eimeria tenella pyruvate kinase and a lower identity of less than 25% to the pyruvate kinases from other organisms. Southern blot analysis indicated that the pyruvate kinase gene existed as a single copy in the T. gondii tachyzoite. The active recombinant enzyme contained four subunits and produced a strongly sigmoid saturation curve with phosphoenolpyruvate as the variable substrate. Fructose 1,6-diphosphate, a general activating factor of pyruvate kinase in most species, did not affect the enzyme activity. However, glucose 6-phosphate radically activated the enzyme. Fructose 2,6-diphosphate suppressed the reaction velocity at a higher concentration of phosphoenolpyruvate. These properties indicate that pyruvate kinase activity in T. gondii is regulated by unusual phosphorylated sugars. PMID- 12632163 TI - A microplate assay for Leishmania amazonensis promastigotes expressing multimeric green fluorescent protein. AB - Convenient and economical assays capable of screening many compounds are vital to advance the development of drug therapy. This is particularly important for many of the infections that occur mainly in the Third World. The development of such a spectrofluorometric assay for the protozoan parasite Leishmania is presented here. Using multimeric (four monomers) green fluorescent protein (GFP), Leishmania amazonensis promastigotes were generated with brightness measurable in 96-well microtiter plates. The promastigotes maintained the parental characteristics, were infective to murine macrophages and to mice, and the level of GFP fluorescence corresponded to the number of inoculated cells. The feasibility of using this assay for testing drugs kinetically and in a concentration-dependent manner, under microplate culture condition, was demonstrated with amphotericin B and the herbicide oryzalin, respectively. This assay is the first to allow a real-time analysis of antileishmanial agents with live promastigotes. The method of expressing multimeric GFP for in vitro drug screening is likely to be extendable to many species of parasitic protozoa. PMID- 12632164 TI - Parasite communities of freshwater fish under flood conditions. AB - The parasite communities of the freshwater fish species roach ( Rutilus rutilus), bitterling ( Rhodeus sericeus amarus) and perch ( Perca fluviatilis) were investigated during a major flood. Differences in parasite community structure due to changes in the host environment were expected. We therefore tested for differences in parasite species abundance and diversity as a consequence of the flood. Potential changes in parasite community structure due to seasonal influences were also considered. We found differences in the composition of parasite species and the proportion of ecto- and endoparasites in the three host species. The parasite community of roach was richer in species than those of perch and bitterling. Roach harboured more ectoparasite than endoparasite species. Parasite communities of both perch and bitterling were richer in endoparasite species and had a lower number of ectoparasite species. Parasite communities of the three different host species responded in different ways to environmental change. Considering all parasite species, no effect of either season or flood was found on the species diversity in either perch or bitterling. The flood caused an increase in the parasite species diversity in roach. For roach and perch, higher parasite abundance were found before the flood compared to after flood. However, no difference was found between the different seasons after the flood. By contrast, bitterling had the highest parasite abundance in the season after the flood. When analysing ecto- and endoparasites separately, a decrease in ectoparasite abundance was found in roach and perch, probably related to the flood, and due mainly to a decrease in monogenean species. A significant increase in the abundance of endoparasites was found in perch. In bitterling, the post-flood increase in ectoparasites was due to a high abundance of Gyrodactylus spp. We conclude that the parasite communities of the three fish species responded in different ways to the flood. This may be due to changes in the host life history strategy or in the immune response in conditions of stress. PMID- 12632165 TI - Kinetics of Taenia solium antibodies and antigens in experimental taeniosis. AB - Two groups of hamsters were infected with Taenia solium cysticerci, one of which was suppressed with methyl-prednisolone acetate on the day of infection and every 14 days thereafter. The other did not receive steroid treatment. Faecal and serum samples were taken prior to infection and then at weekly intervals. Parasite circulating- and coproantigens were detected by a capture ELISA with rabbit polyclonal antibodies against T. solium tapeworms. IgG antibodies in serum and in faecal supernatants were detected by ELISA with excretory-secretory products of T. solium adults recovered from hamsters. Infections remained up to 17 weeks in suppressed hamsters, but after week 11 no tapeworms were found in non-suppressed hosts. T. solium coproantigens in both groups of hamsters were positive from the 1st week post-infection (wpi) until the tapeworms were rejected. Circulating antigens were detected only in non-suppressed hamsters from the 3rd wpi until 1 week before T. solium was eliminated. All infected hamsters developed serum IgG antibodies against tapeworms which were detected from the 2nd wpi and decreased slowly after T. solium expulsion. Specific IgG in faecal supernatants was detected from the 3rd wpi only in non-suppressed hamsters. When suppression was stopped, coproantibodies could also be detected. The presence of IgG antibodies indicates that tapeworms induced an immune response in the experimental host and that when hamsters were suppressed with corticosteroids the immune response was impaired and did not allow the detection of IgG coproantibodies. This indicates, in addition, that the passage of T. solium antigens from the small intestine to the circulation was blocked. PMID- 12632166 TI - Microhabitat distribution of Pseudodactylogyrus anguillae and Pseudodactylogyrus bini among and within gill arches of the European eel (Anguilla anguilla L.). AB - The outlying mean index (OMI) method was used to test for seasonal microhabitat partitioning in Pseudodactylogyrus anguillae and Pseudodactylogyrus bini communities parasitising European eel ( Anguilla anguilla L.) in four seasons. No differences in the occurrence of parasites were found in different seasons or on different gill arches. The OMI analysis showed that hemibranch separation and gill segments were the main factors explaining the variation in parasite niche occupation for the first factorial axis, which was generally more significant than the second factorial axis. The OMI and tolerance indices were compared for both species in the seasons studied, and the main differences between species indices were recorded. P. anguillae and P. bini communities were partitioned in the seasons with high epidemiological parameters of the infection. In the seasons with low epidemiological parameters, or in seasons when the parasite species preferred gill arches, the characters of gill division (lower level of gill division than gill arches) were not important for the partitioning of the Pseudodactylogyrus communities. PMID- 12632167 TI - Hymenolepis pseudodiminuta Tenora et al. 1994 from Apodemus speciosus and H. diminuta: a comparison of experimental infections in rats. AB - The successful maintenance of Hymenolepis pseudodiminuta, isolated from Apodemus speciosus, is described for the first time. In the laboratory, the flour beetle, Tribolium confusum, and F344 rats could serve as intermediate and definitive hosts, respectively. In single worm infections with H. pseudodiminuta, which were carried in two groups of rats, adult worms were recovered from eight and seven out of ten rats, respectively, while Hymenolepis diminuta was found in all of ten rats 6 weeks after inoculation. The worm weight of H. pseudodiminuta in rats was significantly lower than that of H. diminuta. The egg output of H. pseudodiminuta occurred significantly earlier than that of H. diminuta. The number of eggs in the faeces of H. diminuta-infected rats was approximately twofold higher than the number in the faeces of H. pseudodiminuta-infected rats throughout the course of the infection. Mucosal mast cells in rats infected with H. pseudodiminuta were significantly more common than in rats infected with H. diminuta. No detectable IgE antibodies were found in the uninfected and H. diminuta-infected rat groups; however total IgE was detected in H. pseudodiminuta-infected rats but the concentrations were variable between individuals. PMID- 12632169 TI - Ultrastructure of smooth muscle, gap junctions and glycogen distribution in Taenia solium tapeworms from experimentally infected hamsters. AB - Taenia solium adults were grown in hamsters infected by feeding them with cysticerci from pig carcasses. Viable strobilae were collected from the hamster duodenum 20-60 days post-infection, fixed and processed for transmission electron microscopy (TEM). Fourteen strobilae were cut into pieces and embedded in individual blocks. Sections, stained with toluidine blue, were then photographed by light microscopy. Over 1,200 TEM images were obtained from selected blocks. Maturing proglottids exhibited a dense myofilament lattice of connecting fibers, each contained in sarcoplamsic extensions of myocytons and emitting cytoplasmic processes loosely attached to other cells, structures characterized as myocyton myofilament-pseudopod units, which are interpreted as structures involved in the transport of cells and membrane-bound-glycogen from the germinative tissues to mature proglottids. Densely packed membrane-bound glycogen particles were found between the tegumentary cytons of the neck tissue, and as single-stranded particles between the tegumentary cytons of mature proglottids. These were wrapped around cell bodies in the parenchyma of maturing proglottids and as thin cytoplasmic strands between the testicular lobules of mature proglottids. A large number of cell-to-cell adhesions were identified as gap junctions connected to glycogen strands. We suggest that these are involved in the transport of glucose to differentiating tissues. PMID- 12632168 TI - Characterization of sialidase from Entamoaeba hystolitica and possible pathogenic role in amebiasis. AB - Sialidase from Entamoeba histolytica was purified to apparent electrophoretic homogeneity by chromatography on hydroxyapatite, reactive brown agarose, fetuin/agarose and by fast performance liquid chromatography on a MonoQ column. The enzyme had a molecular mass of 65 kDa, as determined by SDS-PAGE. It had an optimum pH of 5.5 and was maximally active at 37 degrees C. It required Ca(2+) and Mg(2+) for activation but was strongly inhibited by Cu(2+), Fe(2+) and Zn(2+). The E. histolytica sialidase exhibited high specificity for Neu5Acalpha2,3lac and methylumbelliferyl-Neu5Ac (4-MU-Neu5Ac) with K(m) values of 0.144 mM and 0.059 mM, respectively. The enzyme was not active against colomic acid and the gangliosides GM(1) and GD(1). It was activated in the presence of lactose and galactose, but was unaffected by glucose, sucrose and mannose. The enzyme was inhibited competitively by 2,3,didehydroneuraminic acid and para-nitro phenyloxamic acid with inhibition binding constants ( K(i)) of 30 micro M and 185 micro M, respectively. The motility of intact E. hystolytica cells was enhanced about 6-fold in the presence of 0.05-0.1 mM Neu5Acalpha2,3lac, 4-MU-Neu5Ac and fetuin. However, the motility of the parasite was highly diminished when incubated with Neu5Acalpha2en and sialic acid-containing compounds. Lysed E. histolytica trophozoites were found to lack neuraminic acid. PMID- 12632170 TI - Molecular karyotypes of Eimeria tenella resolved by PFGE: an evaluation of different agaroses. AB - Molecular karyotypes that revealed size polymorphisms between homologous forms of chromosomes 1, 2, 3 or 4 in three strains of the coccidial parasite Eimeria tenella were derived by pulsed field gel electrophoresis (PFGE) using brands of agaroses marketed either specifically for PFGE or for purposes such as protein electrophoresis or the separation of small molecules of DNA. Chromosomes up to approximately 4 Mbp were well resolved in agaroses marketed for non-PFGE purposes and these products clearly provide useful alternatives when supplies of dedicated PFGE agaroses are limited or withdrawn. PMID- 12632171 TI - Extracellular metalloproteinase activity in Phytomonas francai. AB - Extracellular proteolytic activities were detected in Phytomonas francai culture supernatant. A 67-kDa enzyme was purified by ammonium sulfate precipitation and gel filtration in a HPLC system. This proteinase was optimally active at 28 degrees C and pH 5.0; and the use of proteolytic inhibitors indicated that it belongs to the metalloproteinase class. This is the first report on the purification of an extracellular metalloproteinase from a Phytomonas species. PMID- 12632172 TI - In vitro and in vivo studies on the effect of a combination containing 10% imidacloprid and 50% permethrin against Ixodes ricinus ticks. PMID- 12632173 TI - A survey of bacterial diversity in ticks, lice and fleas from Australia. AB - We isolated bacteria from ticks, lice and fleas. Partial small subunit rRNA sequences were obtained for each isolate and the closest matches in the FastA database were determined. These bacteria were mostly Gram-positive (Firmicutes), although representatives from the Proteobacteria (alpha, beta, gamma subdivisions) and CFB group were also isolated. Most of the isolates we found were from genera that were present in most of the ectoparasites studied, but a few genera were restricted to one species of ectoparasite. The most commonly isolated genera were Stenotrophomonas, Staphylococcus, Pseudomonas, Acinetobacter and Bacillus. Species of Bacillus and Proteus, which have biopesticide potential, were found in some of these ectoparasites. Overall, the communities of bacteria were similar to those found in other studies of parasitic arthropods. PMID- 12632174 TI - Proboscipedia represses distal signaling in the embryonic gnathal limb fields of Tribolium castaneum. AB - Orthologs of the Hox genes Sex combs reduced ( Scr) and proboscipedia ( pd) are active in the developing labial appendages of all insect species tested. The remarkable variation among insect gnathal structures, particularly in the distal podomeres, suggests two Hox genes may enhance the adaptive potential of gnathal appendage morphology. Functional studies in the fruitfly Drosophila melanogaster, the flour beetle Tribolium castaneum and the milkweed bug Oncopeltus fasciatus show that cooperation between Scr and pb has been generally conserved, but specific mechanisms have been altered during evolution. Cross-regulation of pb by Scr is evident in Drosophila and Tribolium, the more closely related of the three species, but not in Oncopeltus. In all three species, pb function is restricted to the distal podomeres, but details are only known for Drosophila and Oncopeltus, two species exhibiting specialized stylate-haustellate mouthparts. Drosophila pb is required for distal Scr expression, and to repress the appendage patterning genes dachshund and Distal-less ( Dll). Oncopeltus pb has the novel capacity to specify leg fates. Little is known about distal functions of Tribolium pb. Hypomorphic mutations of the Tribolium pb ortholog maxillopedia can be arranged in a graded phenotypic series of palp to leg transformations along both the proximodistal and dorsoventral axes. Mid-embryonic expression profiles of Tribolium pb, Scr, wingless ( wg) and Dll genes were examined in maxillopedia hypomorphic and null mutant backgrounds. Levels of pb and Scr are significantly reduced in the distal appendage field. Tribolium pb therefore positively regulates distal Scr expression, a role that it has in common with Drosophila pb. Tribolium wg is normally down-regulated in the distal domain of the embryonic gnathal appendage buds. It becomes activated distally in maxillopedia hypomorphs. Repression of wg by pb has not been reported in the labial imaginal discs of Drosophila. Alterations of Tribolium Scr and wg expression occur in Dll expressing cells, however, unlike in Drosophila labial imaginal discs, Dll expression appears unaffected in pb hypomorphic backgrounds. We conclude that the Hox genes Sex combs reduced and proboscipedia control an appendage organizer and cell autonomous fate determination during embryonic labial palp development in Tribolium. PMID- 12632175 TI - Snail is required for Delta endocytosis and Notch-dependent activation of single minded expression. AB - In the Drosophila embryo, the mesectoderm corresponds to a single row of cells abutting the mesoderm. It is specified by the expression of the single-minded (sim) gene. The information that precisely positions the sim-expressing cells along the dorso-ventral axis is incompletely understood. Previous studies have shown that Dorsal and Twist activate sim expression in a large ventral domain, while two negative regulators, Snail (Sna) and Suppressor of Hairless [Su(H)], repress sim expression in the mesoderm and neuroectoderm, respectively. Repression by Su(H) is relieved in the presumptive mesectoderm by Notch signaling. In this paper, we show that Sna also has a positive regulatory function on sim expression in the presumptive mesectoderm. This positive effect of Sna depends on the Su(H)-binding sites within the sim promoter, suggesting that Sna regulates Notch signaling. In addition, we find that Delta is endocytosed together with the extracellular domain of Notch. The endocytosis of Delta is only seen in the mesoderm and requires the activity of the sna and neuralized (neur) genes. Interestingly, the Neur-mediated endocytosis of Delta has recently been shown to be sufficient for the non-autonomous activation of Notch target genes in wing imaginal discs. We therefore propose a novel model in which Sna positions the mesectoderm via its dual regulatory activity. In this model, Sna cell-autonomously represses sim expression in the mesoderm and relieves Su(H)-dependent repression in a cell non-autonomous fashion by promoting the Neur-dependent endocytosis of Delta in the mesoderm. PMID- 12632177 TI - Identifying functional cis-acting regulatory modules of the yan gene in Drosophila melanogaster. AB - Yan is a nuclear DNA-binding protein that acts as a general inhibitor of cellular differentiation and proliferation in Drosophila melanogaster. The genetic and biochemical mechanisms required for regulating Yan protein function are well understood, however, the molecular mechanism of yan gene transcriptional regulation has not been fully elucidated. Here we show that the dynamic expression of the yan gene is specified by distinct spatial and temporal cis acting regulatory elements in embryos and larval tissues. Each of these distinct elements is thus capable of replicating vital aspects of endogenous yan gene expression. PMID- 12632178 TI - Possible implication of Hox genes Abdominal-B and abdominal-A in the specification of genital and abdominal segments in cirripedes. AB - The crustaceans cirripedes (barnacles) are characterised by the lack of fully developed abdominal segments at any stage of their life cycle. However, in nauplius larvae of the cirripede Sacculina carcini, we detected five small engrailed stripes in a postero-dorsal region behind the sixth thoracic segment, that we interpreted as a vestigial abdomen. Here, we present additional morphological and genetic data on Sacculina to further characterise this structure. Scanning electron microscopy analysis confirms the existence of a segmented region in this part of the naupliar body. However, at the late naupliar stage, this structure stops its development and degenerates. This region expresses the Hox gene Abdominal-B, which may indicate that it actually corresponds to the posterior-most part of the Sacculina trunk. In addition, Abdominal-B expression differentiates two types of larvae that probably correspond to male and female larvae, respectively. In contrast, no abdominal-A expression can be detected in the vestigial abdomen. We discuss the possible implication of the loss or divergence of the Abdominal-A protein in the impaired development of abdominal segments in cirripedes. PMID- 12632176 TI - Evolutionary convergence in Otx expression in the pentameral adult rudiment in direct-developing sea urchins. AB - Convergence is a significant evolutionary phenomenon. Arrival at similar morphologies from different starting points indicates a strong role for natural selection in shaping morphological phenotypes. There is no evidence yet of convergence in the developmental mechanisms that underlie the evolution of convergent developmental phenotypes. Here we report the expression domains in sea urchins of two important developmental regulatory genes ( Orthodenticle and Runt), and show evidence of molecular convergence in the evolution of direct developing sea urchins. Indirect development is ancestral in sea urchins. Evolutionary loss of the feeding pluteus stage and precocious formation of the radially symmetric juvenile has evolved independently in numerous sea urchin lineages, thus direct development is an evolutionary convergence. Indirect developing species do not express Otx during the formation of their five primordial tube feet, the ancestral condition. However, each direct-developing urchin examined does express Otx in the tube feet. Otx expression in the radial arms of direct-developing sea urchins is thus convergent, and may indicate a specific need for Otx use in direct development, a constraint that would make direct development less able to evolve than if there were multiple molecular means for it to evolve. In contrast, Runt is expressed in tube feet in both direct- and indirect-developing species. Because echinoderms are closely related to chordates and postdate the protostome/deuterostome divergence, they must have evolved from bilaterally symmetrical ancestors. Arthropods and chordates use Otx in patterning their anterior axis, and Runt has multiple roles including embryonic patterning in arthropods, and blood and bone cell differentiation in vertebrates. Runt has apparently been co-opted in echinoderms for patterning of pentamery, and Otx in pentameral patterning among direct-developing echinoids. The surprisingly dynamic nature of Otx evolution reinvigorates debate on the role of natural selection vs shared ancestry in the evolution of novel features. PMID- 12632179 TI - An endogenous peptide is involved in internal control of metamorphosis in the marine invertebrate Cassiopea xamachana (Cnidaria: Scyphozoa). AB - In the metagenetic life-cycle of the scyphozoan Cassiopea xamachana metamorphosis of planula-larvae or larva-like buds to polyps is triggered by specific external cues which are transmitted inside the larva or bud where internal signals finally coordinate the initiation of metamorphosis. This study deals with an endogenous metamorphosis inducer present in planulae and buds of Cassiopea. The inductive cue is localized in the basal part of the buds and can be characterized as a peptide with an apparent molecular weight of about 7,000 Da. Further purification was performed via reversed phase HPLC on a C18 column. Additional inhibitor assays revealed that protein kinase C and PI3 kinase, two known elements of the metamorphosis-inducing signal transduction cascade in Cassiopea, may act downstream of the endogenous inducing peptide. PMID- 12632180 TI - AmphiFoxQ2, a novel winged helix/forkhead gene, exclusively marks the anterior end of the amphioxus embryo. AB - A full-length FoxQ-related gene (AmphiFoxQ2) was isolated from amphioxus. Expression is first detectable in the animal/anterior hemisphere at the mid blastula stage. The midpoint of this expression domain coincides with the anterior pole of the embryo and is offset dorsally by about 20 degrees from the animal pole. During the gastrula stage, expression is limited to the anterior ectoderm. By the early neurula stage, expression remains in the anterior ectoderm and also appears in the adjacent anterior mesendoderm. By the early larval stages, expression is detectable in the anteriormost ectoderm and in the rostral tip of the notochord. AmphiFoxQ2 is never expressed anywhere except at the anterior tip of amphioxus embryos and larvae. This is the first gene known that exclusively marks the anterior pole of chordate embryos. It may, therefore, play an important role in establishing and/or maintaining the anterior/posterior axis. PMID- 12632182 TI - Pharmacology of inositol trisphosphate receptors. AB - In almost all cells, cytosolic Ca(2+) is a crucial intracellular messenger, regulating many cellular processes. In non-excitable as well as in some excitable cells, Ca(2+) release from the intracellular stores into the cytoplasm is primarily initiated by the second messenger inositol 1,4,5-trisphosphate (IP(3)), which interacts with the IP(3) receptor (IP(3)R), a tetrameric intracellular Ca(2+)-release channel. This review focuses on the pharmacological modulation of the various functionally important sub-domains of the IP(3)R, including the IP(3) binding domain, calmodulin-binding sites, adenine nucleotide-binding sites and the sites for interaction for FK506-binding proteins and other regulators. We will particularly focus on the pharmacological tools that interfere with these domains and discuss their relative specificity for the IP(3)R, thereby indicating their potential usefulness for unraveling the complex functional regulation of the IP(3)R. PMID- 12632183 TI - The role of endogenous angiotensin II in ischaemia, reperfusion and preconditioning of the isolated rat heart. AB - We examined the possibility that endogenous angiotensin II (AII) is involved in the regulation of the cardiac Na(+)/H(+) exchanger (NHE1) during ischaemia, reperfusion and preconditioning. Mechanical function and intracellular sodium ([Na(+)](i)) were studied in isolated, perfused rat hearts. To test whether AII production might underlie the increased activity of NHE1 on reperfusion, we applied the AII receptor antagonist losartan during ischaemia and reperfusion. Losartan significantly improved mechanical performance on reperfusion and reduced the peak [Na(+)](i) on reperfusion. It has been proposed that preconditioning inhibits the activity of NHE1 in early reperfusion. To test whether this might be because of impaired action of AII on NHE1 we applied AII throughout ischaemia and reperfusion in preconditioned hearts. AII abolished the improved mechanical recovery caused by preconditioning and the peak [Na(+)](i) on reperfusion was similar to that after ischaemia alone. Addition of the NHE1 antagonist cariporide or losartan simultaneously with AII, reversed the deleterious effects of AII on the preconditioned heart. These studies suggest that AII contributes to the activation of NHE1 in early reperfusion and that part of the beneficial effect of preconditioning may be attributed to the abolition of AII-induced activation of NHE1. PMID- 12632185 TI - Effect of migratory cycle and 17beta-estradiol on splenic leukocyte functions in female black-headed gulls. AB - The immune function of wild birds is practically unknown. We have studied several functions of splenic leukocytes from the gull Larus ridibundus. Considering that avian physiology is strikingly affected by the seasonal migratory cycle, those functions were analyzed throughout the seasonal cycle. The functions assayed were: adherence to substrate, chemotaxis and lymphoproliferative response to mitogens. Estrogens have been reported to modulate the immune response in mammals; therefore serum levels of 17beta-estradiol were measured in order to find a putative effect of this hormone on the immune response of female gulls. According to our results, gull splenic leukocytes showed a response similar to that reported in mammals and poultry. In regard to the effect of the seasonal migratory cycle, when the gulls arrived from the migratory flight they showed an immunosuppression as we have obtained in chemotaxis and proliferations assays. The immune response increased during the resting period and especially in the premigratory period. We have found a lack of a direct effect of the age of the animals or even the serum estrogen levels in the immune response of the migratory gulls. These results suggest the existence of other hormones and releasing factors, which would modulate the immune response in gulls, or a unique major effect due to the long flight effort. PMID- 12632184 TI - Calcitonin inhibits proton extrusion in resorbing rat osteoclasts via protein kinase A. AB - Although calcitonin is well known to be a potent inhibitor of bone resorption, it remains unknown how it regulates osteoclastic H(+) transport. In this study, we examined the effects of calcitonin on H(+) extrusion in cultured rat resorbing osteoclasts using an intracellular pH (pHi) indicator, BCECF [2'7'-bis-(2 carboxyethyl)- 5-carboxyfluorescein]. Resorbing osteoclasts were identified by their formation of resorbing pits on calcium phosphate-coated quartz coverslips. Both basal pHi and H(+) extrusion activity were significantly higher compared to non-resorbing osteoclasts. Two types of H(+)-extruding systems were identified by pharmacological and immunocytochemical means: a bafilomycin-A(1)-sensitive and an amiloride-sensitive system [H(+) extrusion mediated by a vacuolar type proton pump (V-ATPase) and by a Na(+)/H(+) exchanger (NHE), respectively]. Calcitonin inhibited both H(+) extrusion activities in a dose-dependent manner and this action was mimicked by protein kinase A (PKA) activators, but not by protein kinase C (PKC) activators. Pretreatment with PKA inhibitors completely suppressed calcitonin-induced inhibition, whereas neither PKC inhibitors nor calcium chelators suppressed it. These results indicate that calcitonin inhibits H(+) extrusion generated by V-ATPase and NHE via PKA activation. These inhibitory mechanisms of H(+) transport by calcitonin are important for the regulation of bone resorption. PMID- 12632186 TI - Ca2+ and voltage dependence of cardiac ryanodine receptor channel block by sphingosylphosphorylcholine. AB - The effect of sphingosylphosphorylcholine (SPC) on the cytoplasmic Ca(2+) and voltage dependence of channel gating by cardiac ryanodine receptors (RyR) was examined in lipid bilayer experiments. Micromolar concentrations of the lysosphingolipid SPC added to cis solutions rapidly and reversibly decreased the single-channel open probability (P(o)) of reconstituted RyR channels. The SPC induced decrease in P(o) was marked by an increase in mean closed time and burst like channel gating. Gating kinetics during intraburst periods were unchanged from those observed in the absence of the sphingolipid, although SPC induced a long-lived closed state that appeared to explain the observed decrease in channel P(o). SPC effects were observed over a broad range of cis [Ca(2+)] but were not competitive with Ca(2+). Interestingly, the sphingolipid-induced, long-lived closed state displayed voltage-dependent kinetics, even though other channel gating kinetics were not sensitive to voltage. Assuming SPC effects represent channel blockade, these results suggest that the blocking rate is independent of voltage whereas the unblocking rate is voltage dependent. Together, these results suggest that SPC binds directly to the cytoplasmic side of the RyR protein in a location in or near the membrane dielectric, but distinct from cytoplasmic Ca(2+) binding sites on the protein. PMID- 12632187 TI - Cholesterol modifies the gating of Kv1.3 in human T lymphocytes. AB - The Kv1.3 potassium channel that belongs to the Shaker family of voltage-gated K(+) channels plays a crucial role in the mitogenic response of T cells. Because it spans the cell membrane its function can be influenced by lipid-protein interactions. In order to study the effect of lipid-protein interactions on the functioning of Kv1.3 we manipulated the membrane cholesterol content in T cells mimicking various physiological conditions by means of the oligosaccharide methyl beta-cyclodextrin (MbetaCD) and its cholesterol-saturated complex (MbetaCD/C). Fluorescence polarization anisotropy and peak current density were used to monitor the efficiency of cholesterol removal (MbetaCD) and loading (MbetaCD/C). Using whole-cell patch-clamp technique we determined the kinetic and steady-state parameters of activation and inactivation of the Kv1.3 currents under different treatment conditions. Upon elevation of cholesterol content by 1 or 1.5 mg/ml MbetaCD/C the rates of both activation and inactivation were slowed. Moreover, the increased cholesterol level in the membrane resulted in a biphasic activation curve. Cholesterol depletion with MbetaCD (0.95 and 1.425 mg/ml) caused no significant changes in the gating characteristics of Kv1.3. The equilibrium between the open and the closed states of the channels was affected by increased cholesterol content, but at the same time steady-state inactivation was unchanged. We argue that manipulation of membrane cholesterol changed both the kinetic properties of Kv1.3 and steady-state parameters of activation by modifying lipid-protein interactions. PMID- 12632189 TI - The serine/threonine kinases SGK2 and SGK3 are potent stimulators of the epithelial Na+ channel alpha,beta,gamma-ENaC. AB - The serum- and glucocorticoid-inducible kinase 1 (SGK1) has been identified as a signalling molecule up-regulated by aldosterone, which stimulates the renal epithelial Na(+) channel ENaC. It is therefore thought to participate in the antinatriuretic action of this hormone. More recently, two isoforms, SGK2 and SGK3, have been cloned. The present study was performed to establish whether SGK2 and SGK3 influence ENaC activity similarly to SGK1. Dual-electrode voltage-clamp experiments in Xenopus laevis oocytes expressing alpha,ss,gamma-ENaC with or without SGK1, SGK2 or SGK3 revealed a stimulatory effect of all three kinases on the amiloride-sensitive current (I(Na)). To establish whether the SGK isoforms exert their effects through direct phosphorylation, we replaced the serine at the SGK consensus site of alphaENaC (alpha(S622A)ENaC) by site-directed mutagenesis. alpha(S622A),beta,gamma-ENaC was up-regulated similar to wild-type ENaC, suggesting that SGK isoforms do not act via direct phosphorylation of the transport proteins. In conclusion, SGK2 and SGK3 mimic the function of SGK1 and are likely to participate in the regulation of ENaC activity. PMID- 12632188 TI - A voltage- and Ca2+-dependent big conductance K channel in cochlear spiral ligament fibrocytes. AB - Evidence is accruing that spiral ligament fibrocytes (SLFs) play an important role in cochlear K(+) homeostasis, but little direct physiological data is available to support this concept. Here we report the presence and characterization of a voltage- and Ca(2+)-dependent big-conductance K (BK) channel in type I SLFs cultured from the gerbil cochlea. A single-channel conductance of 298+/-5.6 pS (n=28) was measured under symmetrical K(+). Membrane potentials for half-maximal open probability (P(o)) were -67, -45 and 85 mV with cytosolic free-Ca(2+) levels of 0.7 mM, 10 microM and 1 microM, respectively (n=8 14). The Hill coefficient for Ca(2+) affinity was 1.9 at a membrane potential of 60 mV (n=6). The BK channel showed very low activity (P(o)=0.0019, n=5) under normal physiological conditions, suggesting a low resting intracellular free [Ca(2+)]. Pharmacological results fit well with the profile of classic BK channels. The estimated half-maximal inhibitory concentration and Hill coefficient for tetraethylammonium were 0.086+/-0.021 mM and 0.99, respectively (n=4-9). In whole cell recordings, the voltage-activated outward K current was inhibited 85.7+/-4.5% (n=6) by 0.1 microM iberiotoxin. A steady-state kinetic model with two open and two closed stages best described the BK gating process (tau(o1) 0.23+/-0.08 ms, tau(o2) 1.40+/-0.32 ms; tau(c1) 0.26+/-0.09 ms, tau(c2) 3.10+/-1.2 ms; n=11). RT-PCR analyses revealed a splice variant of the BK channel alpha subunit in cultured type I SLFs and freshly isolated spiral ligament tissues. The BK channel is likely to play a major role in regulating the membrane potential of type I SLFs, which may in turn influence K(+) recycling dynamics in the mammalian cochlea. PMID- 12632190 TI - Kv channel subunits that contribute to voltage-gated K+ current in renal vascular smooth muscle. AB - The rat renal arterial vasculature displays differences in K(+) channel current phenotypes along its length. Small arcuate to cortical radial arteries express a delayed rectifier phenotype, while the predominant Kv current in larger arcuate and interlobar arteries is composed of both transient and sustained components. We sought to determine whether Kvalpha subunits in the rat renal interlobar and arcuate arteries form heterotetramers, which may account for the unique currents, and whether modulatory Kvbeta subunits are present in renal vascular smooth muscle cells. RT-PCR indicated the presence of several different Kvalpha subunit isoform transcripts. Co-immunoprecipitation with immunoblotting and immunohistochemical evidence suggests that a portion of the K(+) current phenotype is a heteromultimer containing delayed-rectifier Kv1.2 and A-type Kv1.4 channel subunits. RT-PCR and immunoblot analyses also demonstrated the presence of both Kvbeta1.2 and Kvbeta1.3 in renal arteries. These results suggest that heteromultimeric formation of Kvalpha subunits and the presence of modulatory Kvbeta subunits are important factors in mediating Kv currents in the renal microvasculature and suggest a potentially critical role for these channel subunits in blood pressure regulation. PMID- 12632191 TI - Olfactory-hypoglossal connections. AB - Natural olfactory stimulation with amyl acetate significantly modulates the electrical activity of hypoglossal neurons and the electromyographic responses of the tongue musculature. The aim of the present study was to identify and characterize, using neuroanatomical and neurophysiological approaches, the pathways involved in the transmission of the olfactory information to the hypoglossal nucleus (XIIn). The neuroanatomical findings provided the initial demonstration that olfactory information is conveyed from the olfactory bulb to the hypoglossal nucleus via the interpeduncular nucleus (IPn) by both fast disynaptic and different polysynaptic pathways. The latter, in particular, involve many of the brain structures that process olfactory information. The electrophysiological studies demonstrated that the IPn neurons respond with a variety of patterns to natural stimulation of the olfactory receptors, thus supporting the hypothesis that the IPn is a crucial relay station for the elaboration and transmission of olfactory stimuli to XIIn. PMID- 12632193 TI - Compliance changes of the series elastic component of elbow flexor muscles with age in humans. AB - The purpose of this study was to determine the compliance of the series elastic component (SEC) of the elbow flexor muscles in young (n=13, mean age 21.5+/-2.5 years) and elderly (n=15, mean age 67.4+/-4.7 years) subjects. SEC compliance was determined using a quick-release method. Under isometric conditions, myoelectrical activity (EMG) of biceps and triceps muscles was quantified by the root mean square (RMS) value. The compliance index (CI) was defined as the slope of the regression of ln(NC) on ln(NF), where NC and NF are normalised compliance and force respectively. Maximal isometric force and neuromuscular efficiency (torque/RMS) were significantly greater in the young than in the elderly. Antagonist (triceps) co-activation was similar for both groups. The CI after quick-release movements was significantly greater in the young than in the elderly. These results suggest that the SEC compliance of the elbow flexors muscles decreases with age. This decrease in global compliance could be induced by changes both in the active and passive portions of the SEC. These findings may be of functional significance for everyday muscular activity in older people. PMID- 12632192 TI - Effects of extracellular sodium on mu-opioid receptors coupled to potassium channels coexpressed in Xenopus oocytes. AB - Wild-type or mutant H297N or H297Q of the mu-opioid receptor were co-expressed with the inwardly rectifying potassium channel GIRK1 in oocytes from Xenopus laevis. Under voltage clamp, pairs of concentration response curves were generated using the agonist normorphine in a bathing medium containing 38.5 mM sodium or an identical medium in which the sodium was replaced by an equimolar concentration of choline. The maximum currents were greater in the presence of sodium by about 30% at wild-type receptors and by about 100% at the mutant receptors. The EC(50) values tended to increase somewhat as well, though these differences reached statistical significance only for the mutant H297Q. Flame photometry detected no change in the intracellular sodium or potassium concentrations of oocytes, suggesting that the effect of sodium was solely extracellular. Thus sodium, long known for its effects on in vitro ligand binding at mu-opioid receptors, also affects overall transduction as revealed in the Xenopus oocyte model of a complete, living cell system. PMID- 12632194 TI - Rat medial gastrocnemius muscles produce maximal power at a length lower than the isometric optimum length. AB - The interaction of relative muscle length and force-velocity characteristics was investigated in the fully activated rat medial gastrocnemius muscle in situ. Average maximal isometric force (as a percentage of the of the maximal isometric force at L(o,iso)) at relative lengths measured below isometric optimum (L(o,iso)) was 96% at L(o,iso)-2 mm, 88% at L(o,iso)-4 mm and 58% at L(o,iso)-6 mm. Force-velocity curves were obtained at the four relative muscle lengths. There were no significant differences in maximal shortening velocity (approximately 280 mm x s(-1)) between the different muscle lengths. The highest power output (P<0.05) was found at L(o,iso)-2 mm (mean+/-SEM 435+/-19 mW). Peak power values at L(o,iso) (390+/-10 mW) and L(o,iso)-4 mm (395+/-12 mW) were not significantly different, whereas peak power was lowest (P<0.05) at L(o,iso)-6 mm. There was a significant (P<0.01) shift of approximately 1.5 mm in optimum muscle length for force generation during shortening contractions compared with isometric contractions. Shortening velocity had only a minor influence on optimum muscle length for force generation. It is concluded that fully activated muscles produce their maximal power at a length lower than L(o,iso). The difference in optimum length between isometric and dynamic contractions may be related to length-dependent variations in sarcomere length in series during shortening. PMID- 12632195 TI - Variability in fibre properties in paralysed human quadriceps muscles and effects of training. AB - A spinal cord injury usually leads to an increase in contractile speed and fatigability of the paralysed quadriceps muscles, which is probably due to an increased expression of fast myosin heavy chain (MHC) isoforms and reduced oxidative capacity. Sometimes, however, fatigue resistance is maintained in these muscles and also contractile speed is slower than expected. To obtain a better understanding of the diversity of these quadriceps muscles and to determine the effects of training on characteristics of paralysed muscles, fibre characteristics and whole muscle function were assessed in six subjects with spinal cord lesions before and after a 12-week period of daily low-frequency electrical stimulation. Relatively high levels of MHC type I were found in three subjects and this corresponded with a high degree of fusion in 10-Hz force responses (r=0.88). Fatigability was related to the activity of succinate dehydrogenase (SDH) (r=0.79). Furthermore, some differentiation between fibre types in terms of metabolic properties were present, with type I fibres expressing the highest levels of SDH and lowest levels of alpha-glycerophosphate dehydrogenase. After training, SDH activity increased by 76+/-26% but fibre diameter and MHC expression remained unchanged. The results indicate that expression of contractile proteins and metabolic properties seem to underlie the relatively normal functional muscle characteristics observed in some paralysed muscles. Furthermore, training-induced changes in fatigue resistance seem to arise, in part, from an improved oxidative capacity. PMID- 12632196 TI - Energetics of muscular exercise at work onset: the steady-state approach. AB - The energetics of muscular exercise at steady state is tightly coupled to, and dependent on, the events that occurred during the transient phase, of which the steady-state is therefore the "memory". The aim of the present study is to show that it is possible to utilize data obtained at exercise steady state to gain information on variables traditionally assessed during exercise transients. A theoretical model based on the steady-state relationships between mechanical power, O(2) uptake (VO(2)) and phosphocreatine (PC) split allows us to highlight three interdependent parameters: the time constant of the VO(2) on response at the muscular level (tau), the mechanical equivalent of PC splitting and the P/O(2) ratio. The model was applied to experimental data obtained during moderate calf exercise in humans inside an MR unit. For a P/O(2) from 5.6 to 6.2, the obtained tau values range from 10.6 to 24.9 s (for PC concentrations from 17.8 to 37.7 mmol/kg fresh muscle), and the corresponding mechanical equivalents of PC from 9.6 to 10.6 J/mmol. The analysis also shows that the time constant of the O(2) uptake kinetics is strictly dependent on the PC concentration at rest, whereas the mechanical equivalent of PC is unaffected by its concentration. PMID- 12632197 TI - Pulsed local-field fluorescence microscopy: a new approach for measuring cellular signals in the beating heart. AB - In cardiac research, single-cell experimental models have been extensively used to study the molecular mechanisms of intracellular Ca(2+) homeostasis. The results of these studies are usually extrapolated to the tissue level assuming that the phenomena studied at the cellular level are either similar in the intact organ, or only slightly modified by variables that exist at the whole-heart level. The validity of these assumptions has rarely been confirmed experimentally. Common obstacles associated with the study of intracellular Ca(2+) signals in beating hearts include motion artifacts and spatio-temporal limitations of the recording system. In this work, action potentials and intracellular Ca(2+) signals were measured in beating hearts from young rats, with spatio-temporal resolutions similar to cellular studies using a novel pulsed local-field fluorescence technique. This method was based on maximizing emitted fluorescence to increase the signal-to-noise ratio (S/N). The fluorescence emission of the indicator molecules was synchronized with brief (<1 ns), high power (400 W) laser pulses, and the common mode noise of the fluorescence signal was differentially cancelled. To follow rapidly evolving signals, a highly sensitive and fast detection system was used (10 kHz). The spatial resolution was improved using a small (50-200 microm diameter) multimode fiberoptic. Mechanical artifacts were effectively reduced by inserting the fiberoptic into a "floating" glass micropipette sealed to the heart wall with negative pressure. Our results demonstrate that local-field fluorescence microscopy offers an outstanding experimental approach for studying physiological signals at the whole-organ level with the high spatio-temporal resolution common to normal cellular approaches. PMID- 12632198 TI - Molding the ears after anterior scoring and concha repositioning: a combined approach for protruding ear correction. AB - The goal of otoplasty for correction of prominent ears is normalization of shape and position of both pinnas with a certain symmetry. Recreating the antihelical fold and lowering the auriculocephalic angle are the basic processes for achieving a normal-looking ear. Various methods have been described to recreate the antihelical fold. In addition to anterior scoring or abrasion, many involve placement of permanent or absorbable sutures to reform the cartilage to proper shape to correct the deformity. In this study the use of a custom-made silicone mold was described after anterior scoring to recreate the antihelical fold, without using any suture material. Silicone molds provide good shaping of the antihelical fold, scapha, and helical rim while the tissue is healing. Fourteen patients underwent bilateral otoplasty for prominent ears in our clinic over a four-year period. The anthelical fold was created by anterior scoring and the concha-mastoid angle lowered and sutured. After completing the surgical procedure for both ears, custom-made silicone molds were worn by the patient for the first two weeks to maintain proper position. The third week, the patient wore the mold half a day. A head dressing was used for two days and the ears were subsequently supported using a headband. Follow-up ranged from fourteen months to four years. One patient developed a postoperative hematoma in one ear that resorbed spontaneously. Patient satisfaction with the procedure was generally high. Using a shape mold after a combination of anterior scoring and concha repositioning gives predictable results with a natural-looking ear. PMID- 12632199 TI - Resection of the prominent mandible angle with intraoral and external approach. AB - Surgical resection of the prominent mandible angle is a common aesthetic procedure in Asian women. Many females want to change their square-shaped face to a round-shaped face, because the square-shaped face is, not considered attractive in Asian culture. Several approaches have been used for this procedure. Surgeons usually have used the intraoral approach because it can avoid the external scar. However, the intraoral approach can have many drawbacks due to the limited operative field and view. In particular, subcondyle fracture occurs more easily because of the limited motion of the saw. This article presents an operative method that avoids subcondyle fracture and makes the procedure easier to execute. The procedure has two main differences; one is the addition of an external stab incision to the conventional intraoral incision and the other is the use of the reciprocating saw instead of the oscillating saw. Forty-three patients were operated on and we recommend this procedure as an easier procedure with less complication than the intraoral-only approach. PMID- 12632201 TI - Cytoplasmic surface structures of bacteriorhodopsin modified by site-directed mutations and cation binding as revealed by 13C NMR. AB - We have examined how cytoplasmic surface structures of [3-(13)C]Ala-labeled bacteriorhodopsin (bR), consisting of the C-terminal alpha-helix and cytoplasmic loops, are altered by site-directed mutations at the former (R227Q) and the latter (A160G, E166G, and A168G) and by cation binding, by means of displacements of the (13)C NMR peaks of Ala228 and Ala233 (C-terminal alpha-helix), Ala103 (C-D loop), and Ala160 (E-F loop). Cytoplasmic ends of the B and F helices were found to undergo fluctuation motions on the order of 10(-5) s, when such surface structures were disrupted, as viewed from suppressed (13)C NMR signals. This happens also for deionized blue membranes of wild type and A160G, with accelerated fluctuations in the loops. Further, cytoplasmic surface structures of Na(+)-regenerated purple membrane from the blue membrane were significantly modified by Ca(2+) ions up to 1 mM under relatively low ionic strength of 10 mM NaCl, although they are very similar at high ionic strength (100 mM NaCl). To interpret these findings, the following two surface structures were proposed. The C-terminal alpha-helix of the wild type at ambient temperature is involved in a perturbed type, probably tilted toward the direction of the B and F helices, to prevent unnecessary fluctuations of these helices for efficient proton uptake during the photocycle. An unperturbed type of helix is achieved when such a surface structure was disrupted at low temperature or in an M-like state. This view is consistent with previously published data for the "proton binding cluster" consisting of Asp104, Glu166, and Glu234. PMID- 12632203 TI - Magainin 2 channel formation in planar lipid membranes: the role of lipid polar groups and ergosterol. AB - Magainin 2, a polycationic peptide, displays bactericidal and tumoricidal activity, presumably interacting with negatively charged phospholipids in the membrane hosts. In this work, we investigate the role played by the lipid head group in the interactions and self-association of magainin 2 during pore formation in lipid bilayers. Two methods are used: single-channel and macroscopic incorporation into planar lipid membranes. Single-channel incorporation showed that magainin 2 did not interact with zwitterionic membranes, while the addition of negatively charged dioleoylphosphatidylglycerol to the membrane leads to channel formation. On the other hand, magainin 2 did not form channels in membranes made up of dioleoylphosphatidylserine (DOPS), although the addition of ergosterol to DOPS membranes leads to channel formation. This finding could indicate that ergosterol may be a possible target of magainin 2 in fungal membranes. Further support for this hypothesis comes from experiments in which the addition of ergosterol to palmitoyloleoylphosphatidylcholine membranes induced channel formation. Besides the role of negatively charged membranes, this study has shown that magainin 2 also forms channels in membranes lacking heads, such as monoolein and oxidized cholesterol, indicating an interaction of magainin 2 with acyl chains and cholesterol, respectively. This finding provides further evidence that peptide binding and assembly in lipid membranes is a complex process driven by electrostatic and/or hydrophobic interactions, depending on the structure of the peptide and the membrane composition. PMID- 12632200 TI - [(18)F]FMISO and [(18)F]FDG PET imaging in soft tissue sarcomas: correlation of hypoxia, metabolism and VEGF expression. AB - Hypoxia imparts resistance to radiotherapy and chemotherapy and also promotes a variety of changes in tumor biology through inducible promoters. The purpose of this study was to evaluate the use of positron emission tomography (PET) imaging with fluorine-18 fluoromisonidazole (FMISO) in soft tissue sarcomas (STS) as a measure of hypoxia and to compare the results with those obtained using [(18)F]fluorodeoxyglucose (FDG) and other known biologic correlates. FDG evaluates energy metabolism in tumors while FMISO uptake is proportional to tissue hypoxia. FMISO uptake was compared with FDG uptake. Vascular endothelial growth factor (VEGF) expression was also compared with FMISO uptake. Nineteen patients with STS underwent PET scanning with quantitative determination of FMISO and FDG uptake prior to therapy (neo-adjuvant chemotherapy or surgery alone). Ten patients receiving neo-adjuvant chemotherapy were also imaged after chemotherapy but prior to surgical resection. Standardized uptake value (SUV) was used to describe FDG uptake; regional tissue to blood ratio (>or=1.2 was considered significant) was used for FMISO uptake. Significant hypoxia was found in 76% of tumors imaged prior to therapy. No correlation was identified between pretherapy hypoxic volume (HV) and tumor grade ( r=0.15) or tumor volume ( r=0.03). The correlation of HV with VEGF expression was 0.39. Individual tumors showed marked heterogeneity in regional VEGF expression. The mean pixel-by-pixel correlation between FMISO and FDG uptake was 0.49 (range 0.09-0.79) pretreatment and 0.32 (range -0.46-0.72) after treatment. Most tumors showed evidence of reduced uptake of both FMISO and FDG following chemotherapy. FMISO PET demonstrates areas of significant and heterogeneous hypoxia in soft tissue sarcomas. The significant discrepancy between FDG and FMISO uptake seen in this study indicates that regional hypoxia and glucose metabolism do not always correlate. Similarly, we did not find any relationship between the hypoxic volume and the tumor volume or VEGF expression. Identification of hypoxia and development of a more complete biologic profile of STS will serve to guide more rational, individualized cancer treatment approaches. PMID- 12632202 TI - Statistical and fractal analyses of rat prostate cancer cell motility in a direct current electric field: comparison of strongly and weakly metastatic cells. AB - The problems addressed here comprised (1) possible differences in galvanotactic properties of strongly versus weakly metastatic rat prostate cancer cells, with MAT-LyLu and AT-2 as examples, respectively; (2) quantitative description of the responses of the MAT-LyLu cells to direct current (dc) electric fields (EFs) of physiological strength (0.3-3 V/cm); and (3) voltage and time dependency of the cells' responses to the dcEFs. These issues were studied by application of statistical and fractal analyses of the cells' trajectories. The results showed that the MAT-LyLu cells responded strongly to the applied dcEFs by migrating towards the cathode. On the other hand, the galvanotactic response of the AT-2 cells was weak and towards the anode. Further studies of the MAT-LyLu cell motility in dcEFs of increasing strength showed that their response consisted of two voltage domains. Weaker fields (approximately 0.6 V/cm) induced "straightening" of the cells' trajectories without the cells showing a clear tendency to move along the applied field. Stronger fields (>0.6 V/cm) made the cells' movement oriented with respect to the direction of the applied field, without further changing the trajectories' structure. The results also showed that the cells do not perform a directed movement instantaneously after switching on a dcEF of 3 V/cm; approximately 30 min lapsed before the cells were able to fully follow the direction of the applied field. Possible biophysical bases and pathophysiological significance of the results obtained are discussed. PMID- 12632205 TI - Modulation by Zn(2+) and Cd(2+) of GABA(A) receptors of rat cerebellum granule cells in culture. AB - This study aims to characterize more closely the different populations of GABA(A) receptors present on the cerebellar granule cells of the rat. The effects of two divalent cations, Zn(2+) and Cd(2+), on GABA-activated chloride currents were studied using the whole-cell patch-clamp technique. Zinc cations inhibit differently the peak and the steady-state current elicited by 10 micro M GABA. In fact, Zn(2+) appears to be more potent in inhibiting the steady-state component, with a lower IC(50). The inhibition of the peak component is of the competitive type, whereas the inhibition of the steady-state one is mixed, being partly competitive and partly allosteric. In addition, Cd(2+) has an inhibitory effect on GABA-activated chloride currents. In terms of the peak component, its effect is limited in extent with a maximal inhibition of only 26%, but with a high affinity (IC(50) as low as 0.03 micro M). The steady-state component is inhibited by 20% independently from the Cd(2+) concentration, in the 10(-2)-10(2) micro M range. In this case, the inhibitory mechanism appears to be of the competitive type for the peak component and of the allosteric type for the steady-state one. We suggest these data are a further confirmation that the rapidly and slowly desensitizing components of the GABA-activated chloride currents, corresponding respectively to the peak and the steady-state components, are made up of two different receptor populations. PMID- 12632204 TI - Combined nanomanipulation by atomic force microscopy and UV-laser ablation for chromosomal dissection. AB - Nanomanipulation and nanoextraction on a scale close to and beyond the resolution limit of light microscopy is needed for many modern applications in biological research. For the manipulation of biological specimens a combined microscope allowing for ultraviolet (UV) microbeam laser manipulation together with manipulation by an atomic force microscope (AFM) was used. In a one-step procedure, human metaphase chromosomes were dissected optically by the UV-laser ablation and mechanically by AFM manipulation. With both methods, sub-400-nm cuts could be achieved routinely. Thus, the AFM is an indispensable tool for in situ quality control of nanomanipulation. However, already on this scale the dilation of the topographic AFM image due to the tip geometry can become significant. Therefore the AFM images were restored using a tip geometry obtained by a blind tip-reconstruction algorithm. Cross-sectional analysis of the restored image reveals a 380-nm-wide UV-laser cut and AFM cuts between 70 nm and 280 nm. PMID- 12632206 TI - Dynamics of lipid chain attached fluorophore 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD) in negatively charged membranes determined by NMR spectroscopy. AB - We have determined the average location and dynamic reorientation of the fluorophore 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD) attached to a C12 sn-2 chain of a phosphatidylserine (PS) analogue (C12-NBD-PS) in zwitterionic phosphatidylcholine (PC) and negatively charged phosphatidylserine (PS) host membranes. (1)H magic angle spinning nuclear Overhauser enhancement spectroscopy indicates a highly dynamic reorientation of the aromatic molecule in the membrane. The average location of NBD is characterized by a broad distribution function along the membrane director with a maximum indicating the location of the probe in the lipid/water interface of the lipid membrane. This behavior can be explained by a backfolding of the sn-2 chain towards the aqueous phase. Small differences in the distribution profiles of the NBD group along the membrane normal between PC and PS host membranes were found: in a PC host membrane, the NBD distribution has its maximum in the glycerol region; in a PS host membrane, NBD resides mostly in the upper chain region. These differences may be accounted for by packing differences in the PC versus PS host membranes. As seen by (2)H NMR order parameters, PS bilayers show a much higher packing density compared to PC membranes. Consequently, backfolding of the sn-2 chain with the NBD group attached causes a larger decrease of molecular order of the sn-1 chain in PS than in PC membranes. The broad distributions obtained for lipid chain attached NBD molecules reflect the motional freedom and molecular disorder in the liquid crystalline lipid membrane. PMID- 12632207 TI - Electrical capacitance of lipid bilayer membranes of hydrogenated egg lecithin at the temperature phase transition. AB - Electrical capacitance of the planar bilayer lipid membrane (BLM) formed from hydrogenated egg lecithin (HEL) has been studied during many passages through the phase transition temperature. In contrast to the BLM from individual synthetic phospholipids, membranes from HEL did not demonstrate any capacitance change at the phase transition temperature maximum, as measured by differential scanning calorimeter at 52 degrees C. Instead, two temperatures have been discerned by capacitance records: thickening at 42-43 degrees C and thinning at 57-59 degrees C. The first temperature region is close to the transition temperature of dipalmitoyllecithin, whereas the second is close to that of distearoyllecithin, two main components of the HEL. It was suggested that capacitance measurements were able to reveal a phase separation in the BLM from HEL which was not detected by differential scanning calorimetry. The phase transition of the BLM from the liquid crystal state to the gel state is followed by thickening of the bilayer structure, partly due to a gauche- trans transition of lipid molecules but mainly due to redistribution of the solvent n-decane. PMID- 12632208 TI - Nickel inhibits the slowly activating channels of radish vacuoles. AB - The mechanism of inhibition by cytoplasmic nickel of slowly activating channels in radish vacuoles was investigated using the patch-clamp technique. The decrease in the macroscopic current induced by the presence of nickel in the cytoplasmic solution can be described by a Michaelis-Menten equation with an apparent dissociation constant of 0.45+/-0.03 mM. At the single-channel level, nickel moderately decreases the single-channel conductance, since the ratio between the chord conductance in the presence and in the absence of 1 mM cytosolic nickel is 0.89+/-0.06. Experiments performed to study the interaction between calcium, an activator of the channel, and nickel strongly suggest that these two ions bind to two distinct molecular sites. A simple mathematical model predicting the experimental observations is presented. PMID- 12632210 TI - Excitation relaxation in the chemically modified photosynthetic reaction center from Rhodobacter sphaeroides 601. AB - The ultrafast excitation relaxation in the sodium borohydride-treated reaction center of Rhodobacter sphaeroides 601 was investigated with selective excitation. From the femtosecond pump-probe measurement at 790 nm, the excitation relaxation demonstrates a biexponential decay with time constants of about 200 fs and 1.4 ps. By comparison with the result from sodium ascorbate-pretreated modified RS601, it could be concluded that the dynamical trace at 790 nm mainly originates from the contribution of accessory bacteriochlorophyll in the active side, and the electrochromic shift arising from the induced positive charge on the special pair primarily affects the absorption band in the red region of the accessory bacteriochlorophyll in RS601. With direct excitation of the special pair, the charge separation and subsequent electron transfer were observed in borohydride modified RS601. The 2.8 ps component was ascribed to the charge separation and electron transfer from P* to H(A). From the dynamical traces at 790, 800 and 818 nm, the ultrafast energy relaxation from the excited accessory bacteriochlorophyll in the active side is consistent with a two-step energy transfer mechanism. This dynamical observation in modified RS601 is of significance in understanding the physical mechanism of excitation relaxation and energy transfer in the photosynthetic primary process. PMID- 12632209 TI - A consistent potential energy parameter set for lipids: dipalmitoylphosphatidylcholine as a benchmark of the GROMOS96 45A3 force field. AB - The performance of the GROMOS96 parameter set 45A3 developed for aliphatic alkanes is tested on a bilayer of dipalmitoylphosphatidylcholine (DPPC) in water in the liquid-crystalline L(alpha) phase. Variants of the force-field parameter set as well as different sets of simulation conditions or simulation parameter sets are evaluated. In the case of the force-field parameters, the van der Waals constants for the non-bonded interaction of the ester carbonyl carbon and the partial charges and charge group definition of the phosphatidylcholine head group are examined. On the methodological side, different cut-off distances for the non bonded interactions, use of a reaction-field force due to long-range electrostatic interactions, the frequency of removal of the centre of mass motion and the strength of the coupling of the pressure of the system to the pressure bath are tested. The area per lipid, as a measure of structure, the order parameters of the chain carbons, as a measure of membrane fluidity, and the translational diffusion of the lipids in the plane of the bilayer are calculated and compared with experimental values. An optimal set of simulation parameters for which the GROMOS96 parameter set 45A3 yields a head group area, chain order parameters and a lateral diffusion coefficient in accordance with the experimental data is listed. PMID- 12632212 TI - Predominant Bacillus spp. in agricultural soil under different management regimes detected via PCR-DGGE. AB - A PCR system for studying the diversity of species of Bacillus and related taxa directly from soil was developed. For this purpose, a specific 24-bp forward primer located around position 110 of the 16S ribosomal RNA gene was designed and combined with a reverse bacterial primer located at the end of the gene. The specificity of this PCR system for bacilli and related taxons was confirmed on the basis of tests with diverse strains as well as with soil DNA. Analysis of a soil DNA derived clone library showed that the amplified fragments affiliated exclusively with sequences of gram-positive bacteria, with up to 95% of the sequences originating from putative Bacillus species. In particular, sequences affiliated to those of B. mycoides, B. pumilus, B. megaterium, B. thuringiensis, and B. firmus, as well as to related taxa such as Paenibacillus, were obtained. A minority, i.e., less than 6%, of the clones affiliated with other gram-positive bacteria, such as Arthrobacter spp., Frankia spp., and uncultured gram-positives. The amplified fragments were used as templates for a second PCR using bacterial 16S rDNA primers, yielding PCR products of about 410 bp, which were separated by denaturing gradient gel electrophoresis (DGGE). Amplicons indicating Bacillus spp. were found in the gel between 45% and roughly 60% denaturant, whereas those representing other, high-G+C% bacteria, were localized in gel regions with denaturant concentrations exceeding about 60%, thus allowing the distinction between these two groups of sequences. We applied this system to compare the group-specific diversity in bacterial communities in an agricultural soil under different regimes, i.e., permanent grassland, grassland recently turned to arable land, and arable land under agricultural rotation. Differences in the Bacillus related community structures between the treatments were clearly detected. Higher diversities, as judged by Shannon-Weaver indices calculated on the basis of the molecular profiles, were consistently observed in the permanent grassland and the grassland turned into arable land, as compared to the arable land. PMID- 12632211 TI - Diversity of ascomycete laccase gene sequences in a southeastern US salt marsh. AB - The diversity of ascomycete laccase sequences was surveyed in a southeastern US salt marsh using a degenerate primer set designed around copper binding sites conserved in fungal laccases. This gene was targeted for diversity analysis because of its potential function in lignin degradation in the salt marsh ecosystem and because few studies have assessed functional gene diversity in natural fungal communities. Laccase sequences were amplified from genomic DNA extracted from 24 isolates (representing 10 ascomycete species) cultured from decaying blades of Spartina alterniflora, and from DNA extracted directly from the decaying blades. Among the ascomycete isolates, 21 yielded a PCR product of expected size (900 bp) that was tentatively identified as laccase based on sequence similarities to previously published laccase sequences from related organisms. Overall, 13 distinct sequence types, containing 39 distinct sequences, were identified among the isolates, with several species yielding multiple distinct laccase types. PCR amplifications from early and late decay blades of S. alterniflora yielded seven laccase types. Of these, five were composed of sequences >96% similar at the amino acid level to sequences from three cultured ascomycetes previously found to be dominant members of the fungal communities on decaying S. alterniflora blades. Two of the laccase types from the natural-decay clone library were novel and did not match any of the sequences obtained from the cultured ascomycetes. The 39 distinct sequences and 15 distinct laccase sequence types retrieved from the S. alterniflora decay system demonstrate high sequence diversity of this functional gene in a natural fungal community. PMID- 12632213 TI - Life under nutrient limitation in oligotrophic marine environments: an eco/physiological perspective of Sphingopyxis alaskensis (formerly Sphingomonas alaskensis). AB - The oceans of the world are nutrient-limited environments that support a dynamic diversity of microbial life. Heterotrophic prokaryotes proliferate in oligotrophic regions and affect nutrient transformation and remineralization thereby impacting directly on the all marine biota. An important challenge in studying the microbial ecology of oligotrophic environments has been the isolation of ecologically important species. This goal has been recognized not only for its relevance in defining the dynamics of community composition, but for enabling physiological studies of competitive species and inferring their impact on the microbial food web. This review describes the successful isolation attempts of the ultramicrobacterium, Sphingopyxis alaskensis (formerly described as Sphingomonas alaskensis) using extinction dilution culturing methods. It then provides a comprehensive perspective of the unique physiological and genetic properties that have been identified that distinguish it from typical copiotrophic species. These properties are described through studies of the growth phase and growth rate control of macromolecular synthesis, stress resistance and global gene expression (proteomics). We also discuss the importance of integrating ecological and physiological approaches for studying microorganisms in marine environments. PMID- 12632214 TI - The epidemiology of cardiovascular defects, part 2: a study based on data from three large registries of congenital malformations. AB - There were three objectives of this study: to investigate possible specificity in the association between specific cardiac defects and chromosomal anomalies; to evaluate ways of categorizing cardiac defects into larger groups with epidemiological similarities that could indicate similarities in etiology or pathogenesis; and to analyze the relationship between specific cardiac defects and diabetes. We pooled data on infants (aged 1 year or younger) with congenital cardiovascular defects from three large birth defect registries in California, Sweden, and France. The registries in Sweden and France obtained data through reporting from various sources; in California, medical records were reviewed. For severe congenital heart defects, the percentage of infants with identified chromosomal anomalies varied between 0.9% for d-TGV to 68.4% for ECD. In general, specific cardiac conditions have different risk factors. For example, conotruncal defects have been traditionally grouped, but the data presented in this paper indicates more differences for risk factors for the components of conotruncal defects: tetralogy of Fallot, d-TGV, common truncus, and DORV. In general, we suggest the strategy of "splitting" rather than "lumping" when searching for specific genetic factors and/or teratogens. Adequate analysis thus requires large registries or collaboration among registries. The findings did not support constellations between mothers' diabetes and specific defects. PMID- 12632215 TI - The epidemiology of cardiovascular defects, part I: a study based on data from three large registries of congenital malformations. AB - To analyze complex and noncomplex cardiac malformations regarding prevalence and in relation to demographic variables, we pooled data on infants (age 1 year or younger) with congenital cardiovascular defects from three large birth defect registries in California, Sweden, and France. Altogether, 12,932 infants had one or more congenital heart defects out of 4.4 million live births and stillbirths. The registries in Sweden and France obtained data through reporting from various sources; in California, medical records were reviewed. As expected, definitions and ascertained conditions differed among each of the registries. The total rates for severe defects were similar (1.43 per 1,000), but differed for specific defects. Clear differences in epidemiological characteristics existed for specific defects; for example, severe cardiac defects sex ratios were significantly high for hypoplastic left heart syndrome, d-transposition of great vessels, double outlet right ventricle, total anoralous pulmonary venous return, tetralogy of Fallot, and significantly low for pulmonary atresia without ventricular septal defect and endocardial cushion defect. Few defects were similar for several epidemiological characteristics, but, for example, the combination of ventricular and atrial septal defects appeared equivalent with endocardial cushion defect under some circumstances, yet behaved differently with regard to associated noncardiovascular defects. PMID- 12632216 TI - Giant aneurysm of the ductus arteriosus in an infant with pierre-robin sequence. PMID- 12632217 TI - Diaphragmatic hernia. AB - We describe a case of acute dyspnea in a patient with Marfan syndrome secondary to bowel herniation into the thoracic cavity. PMID- 12632218 TI - Heavy sedation versus general anesthesia for pediatric invasive cardiology: a Grayer shade of blue versus a pinker shade of pale? PMID- 12632224 TI - Flow in the early embryonic human heart: a numerical study. AB - Computational fluid dynamic (CFD) experimentation provides a unique medium for detailed examination of flow through complex embryonic heart structures. The purpose of this investigation was to demonstrate that streaming blood flow patterns exist in the early embryonic heart and that fluid surface stresses change significantly with anomalous alterations in fetal heart lumen shape. Stages 10 and 11 early human embryo hearts were digitized as calibrated two dimensional (2D) cross-sectional sequential images. A 3D surface was constructed from the stacking of these 2D images. CFD flow solutions were obtained (steady and pulsatile flow). Particle traces were placed in the inlet and outlet portions of these two stages. Sections of the embryonic heart were artificially reshaped. CFD flow solutions were obtained and surface stress changes analyzed. Streaming was shown to exist, with particles released on one or the other side of the cardiac lumen tending not to cross over and mix with particles released from the opposite side of the cardiac lumen. Shear stress changes (stage 10) occur in the altered lumens. Streaming exists in steady and pulsatile flow scenarios in the embryonic heart models. There are differences in local shear stress distributions with surface shape anomalies of the fetal heart lumen. These observations may help shed light on the potential role of fluid dynamic factors in determining patterns of abnormal heart development. PMID- 12632225 TI - Serial exercise performance in children with surgically corrected congenital aortic stenosis. AB - We examined serial exercise test performance in children with congenital aortic stenosis (AS) treated surgically compared to that of nonoperated children with mild to moderate AS. Maximal treadmill exercise data were assessed in 21 children 5.5 +/- 3.8 years after aortic valve (AO) surgery. Patients had undergone the Ross procedure (n = 6) or previous aortic valvotomy, balloon valvuloplasty, or neonatal aortic valvotomy (n = 15). Follow-up treadmill tests were conducted 3.7 +/- 2.8 years later. Data were compared to those of 19 nonoperated AS patients (mean gradient by echocardiogram <50 mmHg). These patients were exercised 3.6 +/- 3.2 years apart. Endurance time, heart rate, systemic blood pressure, and electrocardiogram were compared as repeated measures between tests and to age- and sex-matched normative data. Postsurgical children with AS had normal endurance times despite low peak heart rates on the initial test, and they maintained endurance over time. Nonoperated children with mean AO gradients <50 mmHg also had normal endurance times on the initial test but increased endurance over 3.6 years. Children with operated and nonoperated AS were able to reach or exceed normal endurance times, which may make it difficult to achieve compliance to imposed activity restrictions in this population. PMID- 12632226 TI - Primary antiphospholipid antibody syndrome and cardiac involvement in a child. AB - We report a case of polyvalvar disease and recurrent thrombosis in a 2-year-old boy due to primary antiphospholipid antibody syndrome. His diagnosis was delayed, and he was treated for other diagnoses, including culture-negative endocarditis. Primary antiphospholipid antibody syndrome is rarely discussed in the pediatric cardiology literature, and this case highlights the need for early diagnosis to prevent recurrent valve dysfunction and thrombosis. PMID- 12632227 TI - Sublingual isosorbide dinitrate-stimulated tilt test for diagnosis of vasovagal syncope in children and adolescents. AB - Vasovagal syncope is the most likely cause of syncope in the young. Head-up tilt table test (HUT) provides the ability to provoke vasovagal syncope under controlled laboratory settings. In adult populations, pharmacologic stimulation with intravenous/sublingual isosorbide dinitrate (ISDN) has been shown to be an alternative to isoproterenol for increasing the diagnostic yield of HUT. In this study, 40 patients aged 9-18 years with unexplained syncope and 12 healthy age matched children were evaluated by HUT to 70 degrees for 45 minutes. If tilting alone did not induce symptoms (syncope and presyncope), 0.1 mg/kg ISDN was given while the patient lay supine. After 5 min, the table was tilted to 70 degrees for 15 min or until the symptoms occurred. The control group consisted of 12 healthy age-matched children studied in a similar manner. Six patients (15%) had a positive basal tilt test. Twenty-five patients (62.5%) lost consciousness following ISDN administration. In the control group, nobody had a syncopal episode during the basal tilt test. However, ISDN administration resulted in 1 positive response (8.3%). The sensitivity of the test was 77.5% and its specificity was 91.6%. It is concluded that sublingual nitroglycerin HUT is suitable for routine clinical practice in children and adolescents with unexplained syncope. PMID- 12632228 TI - Hemopericardium and cardiac tamponade as presenting findings of dexamethasone induced hypertrophic cardiomyopathy complicated by transmural myocardial infarction. AB - This is the first report of dexamethasone-induced left ventricular outflow obstruction causing decreased coronary artery perfusion and apical transmural cardiac infarct that resulted in hemopericardium and rapid cardiac tamponade. The differential diagnosis of hemorrhagic pericardial effusion must now include dexamethasone-induced hypertrophic cardiomyopathy. PMID- 12632229 TI - Reward anticipation, cognition, and electrodermal activity in the conditioned monkey. AB - In the present report, we examine electrodermal activity (skin conductance responses, SCRs) in monkeys trained to perform target-selection (TS) tests. In each test, the animal was presented in successive trials with the same two unequally rewarded targets on a touch screen. The probabilistic contingencies of the rewards associated with each target rendered the selection of the best difficult. Our findings revealed SCRs time-locked to the arm movements toward the rewarded targets, occurring after the target touches. Parameters of the SCRs were stable when the uncertainty of the choices and of the outcomes varied. The results support the hypothesis that the physiological processes indexed by the SCRs are the correlate of anticipatory appetitive behavior. In contrast, there is no evidence that the SCRs reflect cognitive processes associated with the detection of the best target. PMID- 12632230 TI - Effect of accuracy constraint on joint coordination during pointing movements. AB - Given the number of muscles and joints of the arm, more ways are available to produce an identical hand movement when pointing to a target than are strictly necessary. How the nervous system manages these abundant degrees of freedom was the focus of this study of pointing to targets of low and high indices of difficulty (ID). Two essential features of movement synergies were examined. The first reflects the preferred relations among the outputs of each movement element and was studied through principal component analysis. The second feature of synergy reflects the flexibility of those relationships evidenced by the use of multiple, goal-equivalent solutions to joint coordination. This second feature, which is the main focus of this report, was studied using the uncontrolled manifold approach. Motor abundance was defined operationally as the component of variance of joint combinations that left unchanged the value of important performance variables (goal-equivalent variability, GEV). This variance component was contrasted with the component of variance leading to a change in the value of these variables (non-goal-equivalent variability, NGEV). The difference between GEV and NGEV was evaluated with respect to the performance variables movement extent, movement direction, and path of the arm's center of mass. More than 90% of the variance of joint motions across the pointing trial were accounted for by one principal component, indicating a consistent temporal coupling among most joint motions in a single functional synergy. The flexible nature of this synergy was revealed by the variability analysis. All subjects had significantly higher GEV than NGEV for most of the movement path. Thus, variable patterns of joint coordination did not represent noise but the use of equivalent coordinative solutions related to stabilizing important performance variables. Higher GEV than NGEV was present regardless of the task's ID. One exception was at the time of peak velocity, leading to poorer control of movement extent than movement direction. Increasing the task's ID led to an overall reduction of joint configuraion variance, particularly GEV. These results support earlier work indicating that the use of goal-equivalent solutions to joint coordination is a common feature of the control of this and many other motor tasks. Functionally important performance variables appear to be controlled through flexible but task specific coordination among the motor elements. PMID- 12632231 TI - Behavioural characteristics of the quick phase of vestibular nystagmus before and after unilateral labyrinthectomy in guinea pig. AB - The aim of this paper was to characterise the conditions under which the quick phase of vestibular nystagmus is generated in response to sinusoidal horizontal angular accelerations in guinea pig and to determine whether the characteristics of the quick phase are altered following unilateral vestibular deafferentation (UVD). In experiment 1, the quick-phase response to 2-Hz sinusoidal stimuli with different peak head velocities was measured before and after UVD. In experiment 2, lower-frequency stimuli with a fixed amplitude (+/-20 degrees ) were used to measure the eye-movement response. In experiment 1, at 2 Hz, the most noticeable difference between UVD animals and normal animals was a reduction in the number of quick phases generated, particularly when rotating towards the lesioned side: the onset of the quick phase was delayed and occurred at a lower value of peak head velocity compared to normals. However, both these measures probably reflect the depressed slow-phase eye-velocity gain rather than a change in the quick phase mechanism itself, because if a quick phase was generated there was no difference between UVDs and normals on a variety of measures (duration, position, peak eye velocity) for a 2-Hz stimulus. For both UVD and normal animals there did appear to be a position threshold for the onset of a quick-phase eye movement (approximately +/-7.5 degrees ), although it should be noted that the threshold is not an absolute value. In experiment 2, with lower-frequency stimuli, the characteristics of the quick phase itself were altered in UVD animals. At these lower accelerations, although the head velocity and average eye position for the onset of the quick phase did not differ between UVD and normal animals, there were significant differences between UVD and normal animals in the number of quick phases generated (fewer), the duration of the quick phase (longer) and the peak eye velocity of the quick phase (slower). For lower-frequency stimuli there was no evidence of a specific eye-position threshold for the generation of a quick phase, although the position at which a quick phase occurred rarely exceeded the +/-7.5 degrees value obtained in experiment 1. The behavioural data were used to produce a biologically based neural-network simulation of both the slow- and quick-phase components of the vestibulo-ocular reflex, the results of which are presented in a companion paper. PMID- 12632232 TI - A realistic neural-network simulation of both slow and quick phase components of the guinea pig VOR. AB - A realistic neural-network model was constructed to simulate production of both the slow-phase and quick-phase components of vestibular nystagmus by incorporating a quick-phase pathway into a previous model of the slow phase. The neurons in the network were modelled by multicompartmental Hodgkin-Huxley-style spiking neurons based on known responses and projections of physiologically identified vestibular neurons. The modelling used the GENESIS software package. The slow-phase network consisted of ganglion and medial vestibular nucleus (MVN) neurons; the latter were constructed using biophysical models of MVN type A and B neurons. The quick-phase network contained several types of bursting cells which have been shown to have major roles in the generation of the quick phase: burster driver neurons, long-lead burst neurons, pause neurons, excitatory burst neurons and inhibitory burst neurons. Comparison of the output neural responses from the model with guinea pig behavioural responses from the companion paper showed consistency between model and animal data for neuron firing patterns, maximal firing rates, and timing, duration and number of quick phases. Comparisons were made for stable head input and for sinusoidal angular stimuli at a range of frequencies from 0.1 to 2 Hz. Except for data at 0.1 Hz, where the simulation produced one more quick phase per half cycle than the animal data, the number of quick phases was consistent between the model and the animal data. The model was also used to simulate the effects both of unilateral vestibular deafferentation (UVD) and of vestibular compensation after UVD, and the responses in the modelled MVN neurons were affected in a way similar to those measured in guinea pig MVN neurons: the number of quick phases and their timing changed in a similar fashion to that observed in behavioural data. PMID- 12632233 TI - Intermanual transfer of force control is modulated by asymmetry of muscular strength. AB - Interlateral transfer of learning is conceptualized as an index of the degree to which learning takes place at a lower level of motor control, with strong dependence on the effector system, or at a higher effector-independent level in the movement organization hierarchy. In this study, the locus of motor learning was investigated by increasing lateral asymmetry of force between the wrist flexor muscles, and comparing the amount of interlateral transfer of force control in relation to a condition of symmetric force. To perform this contrast, the participants were assigned to one of three groups: symmetric force (SM), who were left with original asymmetries of muscular strength; asymmetric force (AS), who had unilateral training for increment of maximum strength for the wrist flexor muscles; or a control condition (CO). The learning task consisted of launching a small cart across a metallic trackway with the preferred hand, aiming at making the cart achieve an instantaneous velocity of 70 cm/s. This action was practiced for 300 trials by the SM and AS group, while the CO group had active rest. The groups, then, were submitted to a transfer task requiring a mirrored action with the contralateral hand. The results indicated that the SM group achieved significantly higher interlateral transfer of learning as compared to the AS group, which presented response variability similar to the CO group. Analysis of directional trend of error revealed that the AS group presented a significant target overshoot as compared with the symmetric force groups. These findings suggest that an absolute force is learnt at a higher level in the action hierarchy, and that decline in interlateral transfer of learning in the asymmetric force condition was motivated by a resetting in the interplay between higher and lower levels of movement control. PMID- 12632234 TI - Delay improves performance on a haptic spatial matching task. AB - Systematic deviations occur when blindfolded subjects set a test bar parallel to a reference bar in the horizontal plane using haptic information (Kappers and Koenderink 1999, Perception 28:781-795; Kappers 1999, Perception 28:1001-1012). These deviations are assumed to reflect the use of a combination of a biasing egocentric reference frame and an allocentric, more cognitive one (Kappers 2002, Acta Psychol 109:25-40). In two experiments, we have examined the effect of delay between the perception of a reference bar and the parallel setting of a test bar. In both experiments a 10-s delay improved performance. The improvement increased with a larger horizontal (left-right) distance between the bars. This improvement was interpreted as a shift from the egocentric towards the allocentric reference frame during the delay period. PMID- 12632237 TI - Finger control in the tripod grasp. AB - The present study aimed to determine whether grasping is based on either (1) synchronous finger movements producing stereotyped types of grasp, or (2) independently controlled finger movements producing variable final finger postures. Participants reached for and grasped sphere-shaped objects of three sizes. They were allowed to select three different grasp configurations: a "pinch" grip (thumb and index finger), a "middle" grip (thumb and middle finger) and a "tripod" grip (thumb and index plus middle finger). Object distance from the subject was varied in order to verify whether finger control and final finger postures varied according to the degree of accuracy required by target object distance. All the participants always selected the tripod grip when reaching for the large and medium size objects. The pinch grip was used by half of the participants when reaching for the small object, but only in 17% of the trials. Target object distance did not appear to influence the type of selected grip. The tripod grip was found to consist of two different components: an aperture component (opening and closing the gap between the thumb and opposition finger) and a finger separation component (increasing and decreasing the gap between the index and middle fingers). The timing of the aperture component was the same for the index and middle fingers. In contrast, the timing of the finger separation was weakly coupled with the aperture components. Moreover, the relative spatial position among the three fingers during and at the end of grasp varied according to object size. When grasping the large object, both the index finger and the middle finger were in opposition to the thumb. In contrast, when grasping the small object, the index finger was less in opposition to the thumb with respect to the middle finger. The final spatial position of the thumb relative to the starting position was independent of object size, whereas those of the index and middle fingers varied with object size. The results support the notion that grasp is accomplished by using two virtual fingers formed by the thumb and one or more other fingers that synchronously open and close on the object along the opposition space [Arbib 1990; in: Jeannerod M (ed) Attention and performance XIII: motor representation and control. Lawrence Erlbaum, Hillsdale, pp 111-138]. This suggests a degree of coupling between the control of the virtual fingers. PMID- 12632235 TI - Sometimes you see them, sometimes you don't: IPSCs in the rat superficial superior colliculus. AB - Superficial superior colliculus (SSC) neurones were voltage-clamped and the current-voltage relationship of synaptically evoked currents analyzed in vitro. A strong interplay between excitatory postsynaptic currents (EPSCs) and inhibitory postsynaptic currents (IPSCs) was identified. Glutamate receptor antagonists not only fully blocked EPSCs but IPSCs were also frequently reduced by the specific d,l,-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor antagonist (by 66.9%), indicative of glutamate-driven inhibitory projections. The GABA(A )receptor antagonist bicuculline enhanced EPSCs and either abolished or reduced (by 79.3%) IPSCs. The GABA(C) receptor antagonist 1,2,5,6-tetrahydro-(pyridin-4 yl)methylphosphinic acid decreased IPSCs in 80% of cells tested (by 24.1%), but had no effect on EPSCs. Varying the recording conditions influenced postsynaptic currents. At a holding potential of -60 mV, IPSCs were generally produced with intracellular chloride concentrations of both 5 and 10 mM (total n=24/30). However, with perforated-patch recordings using gramicidin, IPSCs were less frequently encountered (n=5/21), suggesting a higher intracellular chloride concentration in a large proportion of SSC neurones. Further assessment of experimental conditions revealed that two frequently used sodium channel blockers, QX-314 (bromide salt, intracellular) or tetrodotoxin (extracellular), shifted the IPSC reversal potential towards more positive values. Hence, IPSCs were not encountered at -60 mV in their presence. The level of stimulation intensity (minimal or maximal) did not influence IPSC production in these conditions. Thus, the current study describes the pharmacological properties of PSCs in the SSC and highlights the impact of experimental conditions on synaptic transmission, which requires consideration for past and present data reported in this preparation. PMID- 12632236 TI - Influence of dynamic tilts on the perception of earth-vertical. AB - The aim of this study was to test the hypothesis that optimal activation of both the semicircular canals and the otoliths provides reliable vestibular cues about self-orientation in space. For this, we measured the ability of subjects to estimate the subjective vertical immediately, 20 s and 90 s after a rapid tilt (180 degrees /s(2)) from upright into different roll positions between 90 degrees left and right side down. Subjects had to estimate the earth-vertical and earth horizontal direction in the dark by (a) setting a luminous line, (b) performing saccades, and (c) verbally declaring body position relative to gravity. The mean error curves from the three paradigms showed consistent E (Muller)- and A (Aubert)-effects, which did not significantly change over time. Horizontal and vertical saccade tasks exhibited different response characteristics, as previously reported by others, which likely reflect different computation mechanisms. The verbal estimation paradigm yielded complementary results to those of the luminous line paradigm and vertical saccade task. The E-effect of the luminous line and the vertical saccade paradigm might be explained by a bias towards earth-vertical due to interactions of vestibular and neck afferent signals. The invariably small A-effect of the luminous line and the vertical saccade paradigm probably reflects somatosensory signals that had relatively weak influence in our experiments. We conclude that phasic activation of the vestibular system reduces the influence of non-vestibular cues observed in low tilt velocity or static experiments. Although this activation generates an E effect, the total error in the range of +/-90 degrees is reduced. PMID- 12632238 TI - Dorsal spinocerebellar tract neurons respond to contralateral limb stepping. AB - Proprioceptive sensory information carried by spinocerebellar tracts provides a major input to the spinocerebellum, which has an important role in coordinating motor output for posture and locomotion. Until recently it was assumed that the information transmitted by the dorsal spinocerebellar tract (DSCT) was organized to represent single muscles or single joints in the ipsilateral hindlimb. Recent studies have shown, however, that DSCT activity represents global kinematic parameters of the hindlimb. We now present evidence that the DSCT neurons are also modulated by passive step-like movements of either hindlimb, implying they receive a bilateral sensory input. About two-thirds of 78 cells studied had significant responses to movements of the contralateral limb alone and about 70% responded differently to bipedal movements than to ipsilateral movement alone. The same basic behavior was observed in anesthetized, paralyzed cats and in unanesthetized, decerebrate cats, although decerebrate cats may have had larger responses on average. The results suggest that many DSCT cells may encode information about interlimb coordination. PMID- 12632239 TI - Coordination of fingertip forces in object transport during locomotion. AB - Walking while carrying a hand-held object requires the generation of appropriate grip forces to offset the inertial forces produced during locomotion. The present study examined the interaction between grip forces and locomotion-induced inertial forces across the gait cycle. Eight subjects transported a container under three conditions: self-paced transport with and without accuracy constraints and a velocity-constrained condition. The results showed that the trunk and transported container moved in a synchronized, sinusoidal pattern during all conditions. Grip and inertial forces of the transporting hand were highly coupled in an anticipatory fashion, regardless of task demands. The inertial forces were higher and the coupling was greater in the faster, unconstrained condition. However, grip force modulation was observed even when the inertial forces acting on the container were small and applied indirectly to the container through the locomotor effects originating in the legs and trunk. We suggest that continuous grip force adjustment is used as a generalized strategy to maximize efficiency during object transport regardless of the size or origin of the inertial forces. PMID- 12632241 TI - Preparatory modulation of the gain of visuo-motor transmission for smooth pursuit in monkeys. AB - Brief movement of a foveated target is known to elicit higher velocity ocular (tracking) responses if the target is in motion rather than stationary. We determined whether similar perturbations of a stationary target would have greater ocular effects if we merely increased the probability that the target might undergo sustained motion. For this, we examined the effect of interleaving trials in which the target was always stationary with trials in which the target underwent sustained motion that required the animal to track. We found that perturbation of the stationary target had a greater effect when there were interleaved trials in which the target moved, as though the gain of the visuo motor transmission had been increased in anticipation of future tracking. PMID- 12632242 TI - Object familiarity affects finger shaping during grasping of fruit stalks. AB - In experiment 1 participants reached and grasped equal green stalks of a green apple and a red strawberry. Stability of the two fruits was equalized by fixing the fruits on the table plane where they were reached for and grasped. The results confirmed that finger shaping was influenced by the size of the fruit body (Gentilucci 2002). Finger shaping was larger when grasping the apple stalk than the strawberry stalk. In addition, deceleration phase lengthened when reaching to grasp the strawberry stalk. In experiment 2 participants reached for and grasped equal wooden matches substituting the stalks of the same apple and strawberry presented in experiment 1. No effect of the fruit body was observed on grasping and reaching the matches. This result excluded that the closeness of the stalk to the fruit body was responsible for the interference effect observed in experiment 1. In experiment 3 participants reached for and grasped equal green stalks of an apple and a strawberry of the same red colour. The same results as in experiment 1 were found. They excluded that a grouping effect due to the same green colour of the stalk and the apple body was responsible for the interference effect observed in experiment 1. Results are discussed as further support of the hypothesis that an object is globally analysed when interacting with it, and familiarity, i.e. the way by which a familiar object is commonly grasped, strongly influences all the other possible interactions with it. PMID- 12632240 TI - Directional asymmetry in smooth ocular tracking in the presence of visual background in young and adult primates. AB - The smooth pursuit system moves the eyes in space accurately while compensating for visual inputs from the moving background and/or vestibular inputs during head movements. To understand the mechanisms underlying such interactions, we examined the influence of a stationary textured visual background on smooth pursuit tracking and compared the results in young and adult humans and monkeys. Six humans (three children, three adults) and six macaque monkeys (five young, one adult) were used. Human eye movements were recorded using infrared oculography and evoked by a sinusoidally moving target presented on a computer monitor. Scleral search coils were used for monkeys while they tracked a target presented on a tangent screen. The target moved in a sinusoidal or trapezoidal fashion with or without whole body rotation in the same plane. Two kinds of backgrounds, homogeneous and stationary textured, were used. Eye velocity gains (eye velocity/target velocity) were calculated in each condition to compare the influence of the textured background. Children showed asymmetric eye movements during vertical pursuit across the textured (but not the homogeneous) background; upward pursuit was severely impaired, and consisted mostly of catch-up saccades. In contrast, adults showed no asymmetry during pursuit across the different backgrounds. Monkeys behaved similarly; only slight effects were observed with the textured background in a mature monkey, whereas upward pursuit was severely impaired in young monkeys. In addition, VOR cancellation was severely impaired during upward eye and head movements, resulting in residual downward VOR in young monkeys. From these results, we conclude that the directional asymmetry observed in young primates may reflect a different neural organization of the vertical, particularly upward, pursuit system in the face of conflicting visual and vestibular inputs that can be associated with pursuit eye movements. Apparently, proper compensation matures later. PMID- 12632243 TI - Event-related desynchronization/synchronization in the putamen. An SEEG case study. AB - Event-related desynchronization (ERD) and synchronization (ERS) were studied during the invasive exploration of an epileptic surgery candidate. An electrode that was targeted in the amygdalo-hippocampal complex passed through the putamen with several contacts. During a simple self-paced motor task, we observed in the putamen a power decline (ERD) in both the alpha and beta frequency bands, and a rebound phenomenon (ERS) in the beta frequency band, concurrent with the movement of each hand. This is the first report of ERD/ERS in the basal ganglia. PMID- 12632244 TI - Positron emission tomographic analysis of dose-dependent NAD-299 binding to 5 hydroxytryptamine-1A receptors in the human brain. AB - RATIONALE: The serotonin 5-HT(1A) receptor has been ascribed a putative role in the pathophysiology and drug treatment of depression. NAD-299 (generic name robalzotan) is a new potential antidepressant with high affinity and selectivity for the 5-HT(1A) receptor. OBJECTIVES: The aim of this positron emission tomography (PET) study was to examine the extent and time-course of 5-HT(1A) occupancy by NAD-299 in the human brain, in relation to plasma concentration after escalating single oral doses. METHODS: Five healthy male subjects received one or more single oral doses of NAD-299 (0.5, 2.5 and 10 mg) in aqueous solution under fasting conditions. Total and unbound (after ultrafiltration) plasma concentrations of NAD-299 were determined by liquid chromatography-mass spectrometry (LC-MC), over a tentative dosage interval of 8 h. Regional 5-HT(1A) receptor occupancy in brain was calculated by the simplified reference tissue model using the radioligand [ carbonyl-(11)C]WAY-100635. RESULTS: After the 10 mg dose, occupancy was high in the raphe (62-85%) and neocortical regions (68-75%) at time for C(max), but had declined considerably (17-44%) at 7 h after dose intake. CONCLUSIONS: This study confirmed that the new selective 5-HT(1A) antagonist NAD-299 occupies 5-HT(1A) receptors in the living human brain in a dose-dependent manner following oral dosage. The curvilinear relationship between NAD-299 drug concentration and 5-HT(1A) receptor occupancy was established and can be used for dose selection in subsequent clinical patient studies. PMID- 12632245 TI - Effect of treatment status on prepulse inhibition of acoustic startle in schizophrenia. AB - RATIONALE: The acoustic startle response is inhibited when the startling stimulus is preceded by a weaker non-startling acoustic stimulus. This phenomenon, termed prepulse inhibition of acoustic startle (PPI), is impaired in schizophrenics compared to normal controls. To date, there is conflicting evidence regarding whether PPI impairments improve with antipsychotic treatment. OBJECTIVES: To examine the effect of medication status on PPI in schizophrenic subjects. METHODS: First, we performed acoustic startle testing on 16 schizophrenic subjects when they were acutely decompensated off medication and later after they were stabilized on antipsychotic treatment. Second, in a between-group design, we tested 21 schizophrenic subjects off medication, 16 subjects on atypical neuroleptics, and 27 subjects on typical neuroleptics. RESULTS: In both the test retest study and the between-group study, ANOVAs revealed no significant changes in startle to pulse alone stimuli, habituation of startle to pulse alone stimuli, PPI, latency to response onset, or latency to response peak between the treatment conditions. CONCLUSIONS: Our results do not support the hypothesis that impaired sensorimotor gating in schizophrenia improves with antipsychotic treatment. Rather, impaired gating persists despite symptomatic improvement on medication. PMID- 12632246 TI - 5-HT(2A) receptor binding is reduced in drug-naive and unchanged in SSRI responder depressed patients compared to healthy controls: a PET study. AB - OBJECTIVE: The aim of the study was to investigate differences in 5-HT(2A) receptor binding between healthy volunteers and patients with major depressive disorder (MDD), either never treated before with antidepressants (drug-naive: DN) or responding to paroxetine treatment (drug-treated: DT). METHODS: Nineteen DN patients with MDD and 15 euthymic DT (paroxetine 4 weeks) patients were compared with a group of 20 healthy controls (C) with positron emission tomography (PET) using [(18)F]fluoroethylspiperone ([(18)F]FESP), a 5-HT(2A) and D(2) receptor antagonist. A "binding index" (BI) of [(18)F]FESP to cortical and basal ganglia regions was calculated as the ratio of the activity in these regions to that of cerebellum. Differences in BI between groups, i.e. C versus DN or DT, were assessed by ANOVA, with or without age as covariate (ANCOVA). RESULTS: A significant reduction in BI ( P=0.003 ANOVA, P=0.001 ANCOVA) was found in DN patients in the frontal, occipital, temporal and cingulate cortices, but not in the striatum. No significant differences emerged between C and DT patients. CONCLUSIONS: The reduction of [(18)F]FESP BI in cortical areas of DN depressed, but not of euthymic DT patients suggests an association between the occurrence of depressive symptoms and impairment of cortical 5-HT(2A) receptors. [(18)F]FESP cortical BI may represent a state marker of depression. PMID- 12632247 TI - Co-release of noradrenaline and dopamine from noradrenergic neurons in the cerebral cortex induced by clozapine, the prototype atypical antipsychotic. AB - RATIONALE: Clozapine has been shown to increase extracellular dopamine (DA) and noradrenaline (NA) in the medial prefrontal cortex (mPFC). A recent study of ours suggested that extracellular DA in the PFC originates not only from dopaminergic but also from noradrenergic terminals, its release being controlled by alpha(2) adrenoceptors. OBJECTIVES: Since clozapine binds to alpha(2)-adrenoceptors, the possibility that it might co-release DA and NA was studied. METHODS: By means of microdialysis coupled to HPLC with electrochemical detection, the effect of clozapine on extracellular DA and NA in the mPFC, densely innervated by DA and NA, was compared to that in the occipital cortex, equally innervated by NA but receiving few DA projections. RESULTS: Extracellular NA was found to be the same in the two cortices, consistent with homogeneous NA innervation. On the other hand, extracellular DA in the occipital cortex was only 29% lower than in the mPFC, in spite of the scarce dopaminergic innervation in the occipital cortex. Clozapine (10 mg/kg IP) increased extracellular DA and NA not only in the mPFC (by about 320% and 290%, respectively) but also in the occipital cortex (by 560% and 230%, respectively). Administration of the alpha(2)-agonist clonidine (0.15 mg/kg) reversed the effect of clozapine in both cortices, while the D(2)-agonist quinpirole (0.1 mg/kg IP) was ineffective. CONCLUSIONS: The results suggest that clozapine, by inhibiting alpha(2)-adrenoceptors, co-releases DA and NA from noradrenergic terminals in the occipital cortex and that the same mechanism might be responsible for the concomitant increase of the two monoamines in the mPFC. PMID- 12632249 TI - Preferential effects of the cannabinoid CB1 receptor antagonist, SR 141716, on food intake and body weight gain of obese (fa/fa) compared to lean Zucker rats. AB - RATIONALE: The selective CB(1) receptor antagonist, SR 141716, has been demonstrated to reduce food consumption in a range of animal species. OBJECTIVE: To assess the effect of chronic administration of SR 141716 on body weight and ingestive behaviour of lean and obese (fa/fa) Zucker rats. METHODS: Lean and obese Zucker rats were orally dosed with SR 141716 (3, 10, 30 mg/kg PO), sibutramine (5 mg/kg PO) or vehicle for one week. Pair-fed controls provided insight as to whether the effect of SR 141716 on body weight was attributable to drug-induced hypophagia. Subsequently, the effect of chronic oral administration of SR 141716 (1, 3, 10 mg/kg) was assessed for 28 days. At the end of this period, all animals were given vehicle for 14 days. The incidence of wet-dog shakes, yawning, scratching, and grooming behaviours, was assessed after acute administration and at weekly intervals thereafter for 4 weeks. RESULTS: SR 141716 dose-dependently decreased food intake and body weight gain in both lean and obese animals. The inhibition of food intake and body weight gain was greater in obese Zuckers than in lean Zucker controls. Changes in the body weights of pair fed controls closely paralleled those of their drug-treated counterparts. Chronic 28-day treatment led to a maintained reduction of body weight gain. Withdrawal of SR 141716 on day 28 resulted in rebound hyperphagia and a significant weight gain. On acute administration, SR 141716 dose-dependently induced motor behaviours that showed tolerance upon repeated administration. CONCLUSION: These data indicate that chronic oral treatment with SR 141716 significantly reduces the food intake and body weight gain of obese and lean Zucker rats, an effect that is greater in obese animals and reversible upon drug withdrawal. PMID- 12632251 TI - Nickel-induced oxidative stress and effect of antioxidants in human lymphocytes. AB - The purpose of this study was to evaluate the oxidative effect in human lymphocytes after acute nickel (Ni) treatment for 1 h; levels of intracellular reactive oxygen species (ROS), lipid peroxidation (LPO) and hydroxyl radicals ((*)OH) were examined in isolated lymphocytes. The potential effects of antioxidants were also examined. After acute treatment, NiCl(2) (0-10 mM) significantly decreased the viability of lymphocytes. NiCl(2) appear to increase the degree of dichlorofluorescein (DCF) fluorescence and the levels of thiobarbituric acid-reactive substances (TBARS) in human lymphocytes in vitro in a concentration-dependent manner. The level of (*)OH was quantified by two main hydroxylated derivates, 2,3- and 2,5-dihydroxybenzate (DHB). Levels of 2,3- and 2,5-DHB were significantly higher in the Ni-treated group than in controls. Catalase partially reduced the NiCl(2)-induced elevation of oxidants and TBARS, whereas superoxide dismutase (SOD) enhanced the level of oxidants and TBARS. Both NiCl(2)-induced fluorescence and LPO were prevented significantly by glutathione (GSH) and mannitol. NiCl(2)-induced increase in generation of (*)OH was prevented significantly by catalase, GSH and mannitol, but not by SOD. These results suggest that NiCl(2)-induced lymphocyte toxicity may be mediated by oxygen radical intermediates, for which the accelerated generation of (*)OH may plays an important role in Ni-induced oxidative damage of human lymphocytes. Catalase, GSH and mannitol each provides protection against the oxidative stress induced by Ni. PMID- 12632250 TI - Stimuli produced by observing responses make rats' ethanol self-administration more resistant to price increases. AB - RATIONALE: Observing responses bring sensory receptors into contact with environmental stimuli. In the observing-response procedure, periods in which an operant response (e.g. pressing a lever) is reinforced by drug deliveries alternate with periods in which this response is never reinforced (i.e. extinction). These alternating periods of drug availability versus extinction are not signaled. Observing responses (i.e. presses on a second lever) produce brief stimuli signaling whether drug is available or not for responses on the first lever. Little is known about how parameters of the drug reinforcer affect drug stimulus observing. OBJECTIVES: The effects of changes in the unit price (responses/reinforcer magnitude) of self-administered ethanol on rats' observing were examined. Also, the effects of an observing-response-produced ethanol stimulus on ethanol consumption were examined by comparing consumption during signaled and unsignaled periods of ethanol availability. METHODS: Rats self administered oral ethanol in the observing-response procedure. The unit price of ethanol in the observing-response procedure was increased by increasing the response requirement for ethanol across conditions. RESULTS: Observing and response rates on the ethanol lever increased and then decreased with increases in the unit price of ethanol. However, ethanol-lever responding and ethanol consumption during periods when ethanol was available were less sensitive to increases in price when the observing-response-produced ethanol stimulus was present. CONCLUSIONS: Observing varies as an orderly function of unit price of a drug reinforcer, and drug stimuli produced by observing responses can make drug consumption less sensitive to increases in price. This procedure may provide an animal model of both attending to drug stimuli and the resultant effects of these stimuli on drug taking. PMID- 12632252 TI - Decreased GSSG reductase activity enhances cellular zinc toxicity in three human lung cell lines. AB - Cellular reduced glutathione (GSH) levels have been identified as an essential determinant in zinc-induced cytotoxicity. However, cytotoxic effects of zinc have also been observed without depletion of GSH stores. In a previous study, the intracellular activity of GSSG reductase (GR) has come into focus (Walther et al. 2000, Biol Trace Elem Res 78:163-177). In the present paper we have tried to address this issue more deeply by inhibiting the activity of cellular GR without any appreciable decreases of cellular glutathione. In three pulmonary cell lines, GR activity was inhibited in a dose-dependent manner by the alkylating agent carmustine (BCNU), a known inhibitor of GR. Cells were pretreated with BCNU for 14 h, followed by exposure to various concentrations of zinc chloride. Then we determined the incorporation of radiolabelled methionine (to assess protein synthesis), and measured the GSH and oxidized glutathione (GSSG) levels. Additionally, GR activity of controls was measured. IC(50) values for zinc induced inhibition of methionine incorporation, as well as GSH contents, was strongly correlated to the decreased GR activity. These results firmly suggest that GR is an important factor in the event chain of zinc cytotoxicity. Together with the results from our previously cited study where impaired regeneration of GSH levels were accompanied by a decrease in total cellular glutathione (GSH + GSSG) we conclude that GSSG itself is an important effector in zinc cytotoxicity. PMID- 12632253 TI - Metabolism of tributyltin and triphenyltin by rat, hamster and human hepatic microsomes. AB - Tributyltin and triphenyltin are metabolized by cytochrome P-450 system enzymes, and their metabolic fate may contribute to the toxicity of the chemicals. In the current study, the in vitro metabolism of tributyltin and triphenyltin by rat, hamster and human hepatic microsomes was investigated to elucidate the metabolic competence for these compounds in humans. The metabolic reaction using microsome NADPH system that is usually conducted was not applicable to in vitro metabolism of organotins, especially triphenyltin. We therefore examined the effects of dithiothreitol (DTT), one of the antioxidants for sulfhydryl groups, to determine the in vitro metabolism of tributyltin and triphenyltin. As a result, the treatment with 0.1 mM DTT in vitro increased the activity of the microsomal monooxygenase system for metabolism of tributyltin as well as triphenyltin; the total yield of tributyltin and triphenyltin metabolites as tin increased, respectively, by approximately 1.8 and 8.9 times for rat, 2.1 and 1.2 times for hamster, and 1.6 and 1.5 times for human. It is suggested that the organotins directly inactivate cytochrome P-450 because of the interaction with critical sulfhydryl groups of the hemoprotein. We confirmed the utility of this in vitro metabolic system using DTT in the hepatic microsomes of phenobarbital (PB) pretreated and untreated hamsters. Thus, the in vitro metabolic system described here was applied to a comparative study of the metabolism of organotins in rats, hamsters and humans. Tributyltin was metabolized more readily than triphenyltin in all the species. In humans, the in vitro metabolic pattern resembled that of hamsters, which were susceptible to in vivo triphenyltin toxicity because of incompetent metabolism. It is possible that the hamster is a qualitatively and quantitatively suitable animal model for exploring the influence of tributyltin and triphenyltin in humans. PMID- 12632254 TI - Expression of cytochromes P450 3A in mouse lung: effects of dexamethasone and pregnenolone 16alpha-carbonitrile. AB - Expression of cytochromes P450 3A (CYP3A) has been reported in the lung, but its regulation has received little attention. In the present study, we assessed lung levels of Cyp3a mRNA, protein and activity in control mice and in mice treated with either dexamethasone (DEX), pregnenolone 16alpha-carbonitrile (PCN) or a mixture of DEX+PCN. Lung expression of the pregnane X receptor (PXR) was also investigated. Constitutive levels of Cyp3a mRNA were found in the lung from control mice by polymerase chain reaction after reverse transcription of total RNA (RT-PCR). These levels were significantly increased (2.0-fold, P<0.05) in mice treated with DEX and further enhanced (2.7-fold increase, P<0.01) in mice treated with DEX+PCN. In control mice, basal levels of Cyp3a protein and activity were also found, as assessed by western blot and measure of testosterone 6beta hydroxylation, respectively. In mice treated with DEX or DEX+PCN, changes in Cyp3a protein and activity exhibited the same pattern as those in Cyp3a mRNA. In contrast, PCN alone failed to trigger consistent increases in lung Cyp3a mRNA, protein and activity. PXR mRNA was not detected in the lung from control or PCN treated mice by RT-PCR, but was found at significant levels in the lungs from mice treated with DEX or DEX+PCN. Our results show that expression of Cyp3a is upregulated by glucocorticoids in mouse lung, and that this effect is potentiated by antiglucocorticoids. This potentiation may involve PXR, expression of which is induced in the lung of glucocorticoid-treated mice. PMID- 12632255 TI - Differential expression of c-fos and c-myc protooncogenes by estrogens, xenobiotics and other growth-stimulatory agents in primary rat hepatocytes. AB - Mechanism(s) of tumour promotion in liver by estrogens and other xenobiotics such as alpha-hexachlorocyclohexane (HCH), 1,1,1-trichloro-2,2-bis(4 chlorophenyl)ethane (DDT) and phenobarbital (PB), are not well understood although it is clear that growth stimulation is one important element in their action. To help in characterizing mechanisms of growth control by these compounds, their effects on the expression of immediate-early protooncogenes c- fos and c- myc have been examined and compared with other compounds that stimulate DNA synthesis in primary cultures of normal rat hepatocytes. Expression of c- fos was undetectable in cultures not exposed to growth factors. Although neither epidermal growth factor (EGF) nor 17beta-estradiol (E(2)) alone had marked effects on c- fos mRNA, the two acted synergistically to cause clear c- fos expression, maximal 1-2 h after growth factor addition and when test agents were added on the first day in culture. Neither insulin nor dexamethasone alone induced c- fos mRNA but stimulation of c- fos expression by EGF plus estradiol occurred earlier in the presence of insulin, and was augmented by preincubation of cells with dexamethasone. EGF + E(2)-induced c- fos mRNA was completely abolished by actinomycin D, suggesting that transcription is the major mechanism for c- fos induction by E(2) + EGF. Compounds that strongly stimulate hepatocyte DNA synthesis such as norepinephrine, pyruvate, prolactin, glutethimide, monensin, ammonium chloride, and normal rat serum when in combination with EGF, all failed (when added with EGF) to affect c- fos mRNA expression. Thus, induction of c- fos expression may be a component of estradiol's growth stimulatory effect in cultured hepatocytes but this is not the case for other compounds that strongly stimulate DNA synthesis. Unlike c- fos mRNA, c- myc mRNA was detectable in hepatocyte cultures without added growth factor, was augmented within 2 h of exposure to EGF, and was further increased by adding E(2), other estrogens or a variety of other stimulators of DNA synthesis in hepatocytes. This suggests that increased c- myc expression may be a common effect of many of these agents in combination with EGF. PMID- 12632256 TI - The anti-breast cancer drug tamoxifen alters Ca2+ movement in Chinese hamster ovary (CHO-K1) cells. AB - The anti-breast cancer drug tamoxifen has recently been shown to cause an increase in intracellular free-Ca(2+) concentrations ([Ca(2+)](i)) in renal tubular cells, breast cells and bladder cells. Because tamoxifen is known to alter ovary function in human patients and in rats, the present study was aimed at exploring whether tamoxifen could alter Ca(2+) movement in Chinese hamster ovary (CHO-K1) cells. Cytosolic free-Ca(2+) levels in populations of cells have been explored by using fura-2 as a fluorescent Ca(2+) indicator. Tamoxifen at concentrations above 1 micro M increased [Ca(2+)](i) in a concentration-dependent manner with an EC(50) value of 8 micro M. The Ca(2+) signal was reduced by removing extracellular Ca(2+), but was not affected by nifedipine, verapamil, diltiazem or ICI 182,780 (an estrogen receptor antagonist). Pretreatment with 1 micro M thapsigargin (an endoplasmic reticulum Ca(2+) pump inhibitor) to deplete the endoplasmic reticulum Ca(2+) abolished 10 micro M tamoxifen-induced Ca(2+) release. Neither inhibition of phospholipase C with 2 micro M U73122 nor depletion of ryanodine-sensitive Ca(2+) stores with 50 micro M ryanodine affected tamoxifen-induced Ca(2+) release. Cell proliferation assays using ELISA revealed that overnight incubation with 5-10 micro M tamoxifen inhibited cell proliferation by 20%, and 20 micro M tamoxifen killed all cells. Together, the results suggest that, in CHO-K1 cells, tamoxifen induced a [Ca(2+)](i) increase by causing store-Ca(2+) release from the endoplasmic reticulum in an phospholipase C-independent manner, and by inducing Ca(2+) influx. The action of tamoxifen appears to be dissociated from estrogen receptor activation. Longer incubation with tamoxifen (>5 micro M) was cytotoxic. PMID- 12632248 TI - Mood, cognition and serotonin transporter availability in current and former ecstasy (MDMA) users. AB - RATIONALE: Chronic recreational ecstasy (MDMA) use has often been reported to be associated with psychopathology, memory impairments and serotonergic alterations. However, the findings have not been consistent. OBJECTIVES: To attempt to replicate these findings, to investigate whether such alterations would be reversible and whether they could be predicted by parameters of previous drug use. METHODS: In a cross-sectional design, 30 current and 31 ex-ecstasy users with ecstasy abstinence of at least 5 months, and 29 polydrug and 30 drug-naive controls were compared on measures of psychopathology, cognitive performance and serotonin transporter availability. RESULTS: The groups did not differ significantly in age, gender distribution, education level and premorbid intelligence. The ecstasy groups did not differ significantly from polydrug controls on most of the relevant parameters of concomitant illegal drug use. Reported drug use was confirmed by hair and urine analyses. All three groups of drug users exhibited significantly elevated psychopathology compared with drug naive controls. Only ex-ecstasy users were significantly impaired on verbal recall. Current ecstasy users showed significantly reduced distribution volume ratios of serotonin transporter availability in the mesencephalon and caudate nucleus. Regression analyses indicated that psychopathology and serotonergic alterations were best predicted by the number of ecstasy tablets taken on a typical event. CONCLUSION: The results indicate that verbal memory impairments were possibly aggravated after prolonged ecstasy abstinence while there was tentative evidence of serotonergic recovery. On the other hand, self-reported elevated psychopathology appeared to be associated with polydrug use in general and not specifically with ecstasy use. PMID- 12632257 TI - Nasal midazolam as a novel anticonvulsive treatment against organophosphate induced seizure activity in the guinea pig. AB - Seizures and status epilepticus, which may contribute to brain injury, are common consequences of exposure to organophosphorus (OP) cholinesterase inhibitors. Effective management of these seizures is critical. To investigate the efficacy of nasal midazolam as an anticonvulsive treatment for OP exposure, as compared to intramuscular midazolam, guinea pigs were connected to a recording swivel for electrocorticograph (ECoG) monitoring and clinical observation. The experimental paradigm consisted of pyridostigmine pretreatment (0.1 mg/kg i.m.) 20 min prior to sarin exposure (1.2x LD(50,) 56 micro g/kg i.m.). One minute post-exposure, atropine (3 mg/kg i.m.) and TMB-4 (1 mg/kg im) were administered. Within 3-8 min after sarin exposure all animals developed electrographic seizure activity (EGSA), with convulsive behavior. Treatment with midazolam (1 mg/kg i.m.) 10 min after the onset of EGSA abolished EGSA within 389+/-181 s. The same dose was not effective, in most cases, when given 30 min after onset. However, a higher dose (2 mg/kg) was found efficacious after 30 min (949+/-466 s). In contrast, nasal application of midazolam (1 mg/kg) was found most effective, with significant advantages, in amelioration of EGSA and convulsive behavior, when given 10 min (216+/-185 s) or 30 min (308+/-122 s) following the onset of EGSA ( P<0.001). Thus, nasal midazolam could be used as a novel, rapid and convenient route of application against seizure activity induced by nerve agent poisoning. PMID- 12632258 TI - Comparison of hepatotoxicity and metabolism of butyltin compounds in the liver of mice, rats and guinea pigs. AB - The hepatotoxicity of tributyltin chloride (TBTC) and dibutyltin dichloride (DBTC) was compared among mice, rats and guinea pigs in vivo. Further, the metabolism of these butyltin compounds in the liver was also investigated in these species. The oral administration of TBTC and DBTC to mice induced obvious liver injury, as demonstrated by both serodiagnosis and histopathological diagnosis. The concentrations of TBTC and DBTC that induced hepatotoxicity in mice at 24 h after oral administration were 180 and 60 micro mol/kg, respectively. In the case of rats, the liver injury induced by TBTC and DBTC was detected at 24 h by the serodiagnosis, but not by histopathological diagnosis. On the other hand, in guinea pigs, TBTC and DBTC administration did not produce any clear liver injury at 24 h, as evaluated by these two diagnostic methods. Thus, the following ranking was obtained with regard to increasing order of sensitivity to liver injury caused by TBTC and DBTC: mice, rats and guinea pigs. The total butyltin contents in the liver of mice were equivalent at 3 h and 24 h after the administration of TBTC or DBTC; however, the contents in the liver of rats and guinea pigs were relatively lower at 3 h and higher at 24 h than those of mice, although there were no differences between rats and guinea pigs in the total liver butyltin content. Concerning the liver metabolism of these butyltin compounds, the main form of butyltin compounds in these animals treated with TBTC was DBTC within 3 h after oral administration, while the main metabolites at 24 h were different in each species, indicating that the liver metabolism of TBTC might vary by animal type. When the animals were treated with DBTC orally, DBTC was hardly metabolized in the livers of these animals even at 24 h, and the liver levels of DBTC were two times greater in mice and guinea pigs than in rats at 3 h and were lower in mice at 24 h than in rats and guinea pigs. The analysis of cellular distributions of DBTC in the liver at 3 h after the administration showed that the levels of DBTC in the nuclear, microsomal and mitochondrial fractions of mice hepatocytes were relatively higher than in those of rats, which were greater than in those of guinea pigs. These results suggest differences in the sensitivity of mice, rats and guinea pigs to hepatotoxicity caused by butyltin compounds and demonstrate that the difference in the sensitivity of these animals to the hepatotoxicity induced by TBTC and DBTC may be partly due to differences in hepatic metabolism of TBTC and in the distribution of DBTC within cell organelles, respectively. PMID- 12632259 TI - Cloning and characterization of a gene cluster involved in tetrahydrofuran degradation in Pseudonocardia sp. strain K1. AB - A gene cluster involved in the utilization of tetrahydrofuran by Pseudonocardia sp. strain K1 was cloned and sequenced. Analysis of a 9.2-kb DNA fragment revealed eight ORFs. The genes designated as thmADBC encode the components of a putative monooxygenase exhibiting a high similarity to different binuclear-iron containing multicomponent monooxygenases. thmA encodes the derived 545-amino-acid oxygenase alpha-subunit, thmD the 360-amino-acid reductase component, thmB the 346-amino-acid oxygenase beta-subunit, and thmC the 117-amino-acid coupling protein. Upstream of the thm genes, an additional ORF ( sad) was identified coding for a protein with high similarity to various aldehyde dehydrogenases. A succinate semialdehyde dehydrogenase activity was specifically expressed in tetrahydrofuran-grown cells. N-terminal sequence analysis of the purified protein revealed that it is encoded by sad. Northern blot analysis indicated that transcription of the thm genes and sad was specifically induced during growth on tetrahydrofuran. Mono-, di- and polycistronic transcripts of these genes were detected. Primer-extension analysis identified transcriptional start sites 37, 61, and 41 bp upstream of the translation start of sad, thmA, and thmB, respectively. Additional ORFs were identified upstream ( orfY) and downstream ( orfZ and aldH) of the thm genes. Furthermore, the data indicated that the analyzed gene cluster was present as a single copy and located on a plasmid. PMID- 12632260 TI - Genome-wide transcriptional changes during the lag phase of Saccharomyces cerevisiae. AB - The set of physiological and metabolic changes occurring immediately after inoculation and during the lag phase is thought to be of vital importance for optimal offset of fermentation. The transcriptional changes taking place during the lag phase after inoculation of a late-respiratory-phase yeast culture into fresh, minimal medium were investigated by use of Yeast GeneFilters. In response to the nutritional up-shift, 240 open reading frames were at least five-fold induced and 122 were at least five-fold repressed. These genes were hierarchically clustered according to their lag-phase expression patterns. The majority of the induced genes were most highly induced early in the lag phase, whereas strong repression generally occurred later. Clustering of the genes showed that many genes with similar roles had similar expression patterns. Repressed genes, however, did not cluster as tightly according to function as induced genes. Genes involved in RNA and protein synthesis and processing showed a peak in expression early in the lag phase, except most ribosomal protein genes, which were induced early and whose expression was sustained. Genes involved in chromatin/chromosome structure showed late induction. The correlation between function and expression pattern for these genes indicates regulation by similar mechanisms. Much of the transcriptional response observed appeared to be due to the presence of glucose in the new medium. PMID- 12632261 TI - Application of crossover-PCR-mediated deletion-insertion mutagenesis to analysis of the bdhA-xdhA2-xdhB2 mixed-function operon of Sinorhizobium meliloti. AB - The bdhA-xdhA2-xdhB2 mixed-function operon was used to demonstrate the application of crossover PCR to the construction of in-frame, non-polar deletion insertion mutations in Sinorhizobium meliloti. Replacement of a 474-bp internal portion of the 774-bp coding sequence of bdhA with a 21-bp in-frame synthetic sequence resulted in loss of the bdhA-encoded d-3-hydroxybutyrate dehydrogenase activity. Such mutants retained the xanthine oxidase activity encoded by xdhA2 xdhB2, thus illustrating the non-polar nature of the mutation. This method of constructing unmarked, in-frame deletions should be generally applicable to functional genomics studies in S. meliloti and other alpha-Proteobacteria. PMID- 12632268 TI - Minimal incision aortic surgery (MIAS). AB - This study evaluates the clinical and economic impact of using less extensive minimal invasive aortic surgery (MIAS) for elective treatment of infrarenal aortic aneurysms (AAA) and aortoiliac occlusive disease (AIOD) in two independent surgical departments. Surgeons from two institutions conducted a prospective consecutive, nonrandomized analysis of MIAS electively performed in 80 patients. MIAS outcomes were compared with 80 consecutive elective standard open aortic procedures (40 from each institution), which were performed during the same time period. Cost analyses for MIAS and standard open repair were performed at each institution. Our results indicated that MIAS is as safe as standard open repair, is more cost-effective, and has significantly shorter hospital stays than with standard open repair. PMID- 12632262 TI - Intermittent flow increases endotoxin-induced adhesion of human erythrocytes to vascular endothelial cells. AB - OBJECTIVE: To investigate the effects of different conditions of flow on endotoxin induced adhesion of human red blood cells (RBC) to human umbilical vein endothelial cells (HUVEC). DESIGN AND SETTING: Prospective, randomized, controlled in vitro study in a university-affiliated cell biology laboratory. SUBJECTS. Human erythrocytes, human vascular endothelial cells. INTERVENTIONS: Superfusion of HUVEC monolayers with human erythrocytes incubated with either saline (CON) or endotoxin (ETX) with different flow pattern (basic flow rates of 0.65 or 1.3 ml/min; intermittent flow, IMF). The CON/0.6, CON/1.3, CON-IMF/1.3 ( n=7/group) groups served as control, and in test groups ETX/0.6, ETX/1.3, ETX IMF/0.6, and ETX-IMF/1.3 ( n=7/group) both RBC and HUVECs were incubated with ETX and flow pattern and rates varied. In the IMF experiments flow rates of 0.65 and 1.3 ml/min were combined with stop-and-go flow pattern. MEASUREMENTS AND RESULTS: At continuous flow of 0.65 ml/min erythrocyte adhesion was 61+/-5 cells/mm(2) in CON and 172+/-25 cells/mm(2) after ETX. When flow rate was increased to 1.3 ml/min, adhesion decreased to 27+/-4 cells/mm(2) in CON and 93+/-18 cells/mm(2) after ETX. IMF conditions had no effect on RBC adhesion of naive RBC but increased the number of adhesive erythrocytes after incubation with ETX both at 0.65 ml/min (287+/-33 cells/mm(2)) and at 1.3 ml/min (148+/-13 cells/mm(2)). CONCLUSIONS: RBC adhesion to vascular endothelium is affected by rate and pattern of blood flow. Higher flow rates or shear forces reduce RBC adhesion while stop and-go flow pattern favored adhesion of ETX-treated erythrocytes to HUVECs. These findings suggest that altered RBCs interact with altered flow patterns potentially contributing to the microcirculatory injury observed in sepsis. PMID- 12632269 TI - Mesenteric Buerger's disease. AB - A young male patient with a history of heavy smoking and low-extremity Buerger's disease was operated on because of small bowel infarction. The postoperative histopathological diagnosis was mesenteric involvement of Buerger's disease. Mesenteric Buerger's disease is extremely rare, the early diagnosis is difficult, and the prognosis is poor. Considering the difficulty in diagnosis and the poor prognosis, patients with Buerger's disease presenting with gastrointestinal manifestations should be carefully evaluated, and early surgical intervention is recommended. PMID- 12632270 TI - Septic embolization to the carotid bifurcation presenting as carotidynia. AB - Carotidynia is a term that is best defined by neck pain associated with tenderness over the carotid artery. The differential diagnosis of this entity is broad and includes vascular disorders such as carotid dissection, aneurysm, occlusion, and arteritis. We describe the first reported case of carotidynia caused by septic embolization to the carotid bulb and offer a logical plan for its management. PMID- 12632271 TI - The circumsphere as a tool to assess distortion in [4Fe-4S] atom clusters. AB - The geometry proposition that "four points not in a plane describe one and only one sphere" provides a novel tool for analyzing protein-induced distortions in [4Fe-4S] clusters. A geometrically perfect reference structure comprises interlaced, regular tetrahedra of Fe, S, and S gamma atoms having T(d) symmetry. Three circumspheres are defined by the three sets of four atoms, the circumcenters of which are unique points within the cluster. The structure is thus re-defined by the positions of the circumcenters in xyz space and the r, theta, phi of each atom on its respective sphere. Analysis of 12 high-resolution structures of protein-bound and small molecule [4Fe-4S](SR)(4) clusters revealed: (a) the circumcenters are generally non-coincident by approximately 0.01 to approximately 0.06 A; (b) the Fe radius, r(Fe), is nominally independent of core oxidation state, having values between 1.66 to 1.69 A, whereas r(S) and r(SG), which have ranges of 2.18-2.24 A and 3.87-3.94 A, respectively, both increase by as much as approximately 3% upon reduction from the 3+ to the 1+ core valence; (c) deviation of some atoms from the theta, phi of a perfect tetrahedron can be large, approximately 10 degrees, and sets of atoms can show patterns of motion on their spheres that result from changes in Fe-S bond lengths. Density functional theory calculations suggest that the [4Fe-4S] core itself requires rather little energy to distort (approximately 2 kcal/mol), whereas significantly more energy is required to distort the Sgamma shell (~4 kcal/mol) to that of cluster I in Clostridium acidurici ferredoxin. PMID- 12632272 TI - The solution structure of the Ga(III)-bleomycin A2 complex resolved by NMR and molecular modeling; interaction with d(CCAGGCCTGG). AB - The solution structure of the Ga(III)-bleomycin A2 complex (GaBLM) has been determined using 2D NMR methods in combination with molecular dynamics calculations. Complete assignment of the amide and amine protons, observation of 80 NOEs and measurement of 15 (3)JH(-H) coupling constants provided us with a well-defined structure using a restrained simulated annealing protocol. On the basis of distance and dihedral angle constraints agreement, along with potential energy considerations, the favored model is a five-coordinate complex with the primary amine of beta-aminoalanine holding the axial position of a distorted tetragonal pyramid. The disaccharide moiety of GaBLM is not a ligand, sharing the same side of the equatorial plane with the axial amine ligand. Titration of the self-complementary oligonucleotide d(CCAGGCCTGG) with GaBLM results in the formation of only one 1:1 complex in slow exchange on the NMR time scale. Our data indicate that the bithiazole moiety intercalates between the C6*G15 and C7*G14 base pairs, in a similar mode to that reported by earlier studies. Structural implications and comparisons to other metallo-bleomycins are discussed. PMID- 12632274 TI - [Laparoscopic appendectomy with mini-instruments]. AB - In the period 01/00-12/01 in 35 out of 199 patients the laparoscopic appendectomy was performed with mini-instruments. This paper gives the prospectively registered data as well as our standardized technique of laparoscopic appendectomy with mini-instruments. The operations were performed via a 12 mm umbilical trocar and two suprapubic mini-trocars. Preparation was done with mini instruments guided by 10 mm/30 degrees optic. Stapling of appendix and ligature of mensenteriolum via the 12 mm umbilical trocar was guided by 30 degrees -mini optics. There was only one change of a mini-trocar to a 10 mm trocar for better view. 33 patients were female, mean age 23.4 years, mean BMI 21.1, mean operation time 41.1 minutes, mean post-op stay 3.3 days. There were no intra- or postoperative complications. The laparoscopic appendectomy with mini-instruments can be done by experienced surgeons in slender patients with low inflammation grade of the appendix without an increased complication rate and with a very good cosmetic result. PMID- 12632275 TI - [Laparoscopic treatment of small bowel obstruction with intussusception, volvulus and appendicitis caused by an inflammatory Meckel's diverticulum]. AB - Indications and contraindications for diagnostic laparoscopy in cases of intestinal obstructions are discussed in connection with a 16-year-old patient's case report with small bowel obstruction including intussusception, volvulus and appendicitis caused by an inflammatory Meckel's diverticulum. The use of diagnostic laparoscopy in cases of intestinal obstructions is controversial. In the described case we performed completely laparoscopically the derevolving and desusception of the volvulus and intussusception, the appendectomy and resection of the diverticulum. The procedure itself and the postoperative course were uneventful. PMID- 12632273 TI - Prospective pilot study of neoadjuvant chemotherapy with 5-fluorouracil, folinic acid and oxaliplatin in resectable liver metastases of colorectal cancer. Analysis of 42 neoadjuvant chemotherapies. AB - PURPOSE: Since there are currently no data available from a prospective trial, the primary objective of this prospective study was to investigate whether the rate of R0-liver resections without morbidity would be at least 50 % in patients with neoadjuvant chemotherapy for colorectal liver metastases. PATIENTS AND METHODS: 42 patients were treated with a biweekly FOLFOX regimen. Chemotherapy consisted of a 2-hour infusion of folinic acid (FOL) 500 mg/m2, followed by a 24 hour infusion of 5- fluorouracil (F) 2000 mg/m2 daily for two days. Oxaliplatin (OX) 85 mg/m 2 was given simultaneously with FOL. Treatment allocation was randomized with either 3 or 6 cycles for the final 30 patients. A liver resection was performed 2 to 5 weeks after the final infusion. RESULTS: An objective response was observed in 20 of 42 patients (response rate was 27 % higher after 6 cycles). Liver resection (R0) could be performed in 34 patients. Postoperative complications were reported in 14 patients (13 occurring within 30 days after resection) and severe complications in 5 cases (including two deaths after extended resection). Liver failure and persistent biliary fistula were the most frequently documented complications. There was no relevant difference in safety criteria between 3 and 6 applications. CONCLUSION: The use of neoadjuvant chemotherapy in resectable liver metastases induced significant remissions without increasing morbidity. The rate of severe complications and cases of no R0 resection in this study was 31 % and was with that significantly lower than 50 % (95 % CI 17.6 %-47.1 %). The risk to the patient is therefore acceptable when undergoing neoadjuvant treatment in a prospective intergroup trial. PMID- 12632276 TI - [Short bowel ileus due to a stenotic ileocecal tumor of tuberculous origin]. AB - Abdominal tuberculosis is a rare disease. Therapy usually consists in drug treatment while surgery is only performed in complicated cases. We report on a 75 year-old man who presented with clinical and radiologic signs of bowel obstruction. The patient complained of weight loss and B-symptoms. There was a history of drug treatment for tuberculosis of the neck and throat in the medical department four weeks prior to admission. Intraoperatively a coecum stenosis was found due to a large tumor that macroscopically appeared with a diffuse carcinomatosis of the peritoneum. A right hemicolectomy was performed. Histopathology findings revealed a granulomatous inflammation with epithelioid cells. Signs of peritoneal carcinomatosis were not seen. The patient recovered well after an additional tuberculostatic treatment and was discharged after 14 days. At present he is free of symptoms. Even after tuberculostatic treatment lymphatic tuberculosis might show signs of progression that can lead to an abdominal presentation. PMID- 12632277 TI - [Complication rate after thoracoscopic and conventional lobectomy]. AB - AIM: To determine whether a thoracoscopic lobectomy used for treatment of primary non-small cell lung cancer shows a higher or lower morbidity in comparison to the conventional resection (thoracotomy) postoperatively. METHODOLOGY: 42 patients with the presumption diagnosis of a lung cancer received a thoracoscopic lobectomy (n=28) or a conventional lobectomy (n=14). Postoperative complication rate (30 days) was analysed prospectively. There were no differences between the groups referring to age, co-morbidity, localisation of the tumor and stage. RESULTS: Patients with a conventional operation revealed a significant higher morbidity in comparison to the patients of the thoracoscopic group (50 % vs. 14.2 %, p=0.03). CONCLUSION: The thoracoscopic lobectomy represents a safe operation method and an alternative to the conventional operation (thoracotomy). PMID- 12632278 TI - [Complex fractures of the humeral head: which long-term results can be expected?]. AB - BACKGROUND: Although improved techniques of internal fixation and prosthetic replacement were introduced successfully in the field of operative fracture care, treatment of complex fractures of the proximal humerus also involving the humeral head remains to be a challenge to the trauma surgeon. We therefore investigated clinical and radiological long-term results after humeral head preserving procedures. METHODS: 41 patients with a three or four part fracture of the humeral head were evaluated on average 6.6 years (min. 4.4 y; max. 9.0 y) after the trauma. The evaluation was based on the Constant- and HSS score clinically and on the Neer score radiologically. Nine patients were treated conservatively (group A), 13 patients had primarily an operative treatment (group B) and another 19 were operated upon after failure of conservative means. 24 of the patients were female and 17 male, with an average age of 52 years (min. 14.4 y; max. 71.2 y). According to Neers 's fracture classification of humeral head fractures we saw 14 type IV, 25 times a combination of type IV and V and in another 2 cases a type VI fracture. RESULTS: In group A (conservative) the Constant score showed on average 82.0 points for the injured and 95.3 points for the contralateral shoulder, the HSS score revealed 73.6 points and Neer's x-ray score 5.6 points. Group B (operated) showed also good results on average according to a Constant score of 72.1 points (fractured humerus) compared with 98.1 points of the contralateral shoulder. HSS score was 64.7 points. The radiological results reached 4.0 points. Group C (conservatively failed, secondary operation) achieved 68.2 points for the injured side and 95.8 points for the contralateral side according to Constant and 59.5 points according to HSS score. The x-ray evaluation showed 5.3 points. Fracture type did not influence the outcome in any of the groups. There was no humeral head necrosis in group A, one in group B (2.4 %) and four in group C (9.8 %). CONCLUSION: These data show that regarding to clinical and radiological long-term results also complex fractures of the humeral head should be treated by head preserving procedures. PMID- 12632279 TI - [Epidemiology, treatment and results of proximal humeral fractures: experience of a district hospital in a sports- and tourism area]. AB - AIM OF THE STUDY: The epidemiology, therapy and results of proximal humeral fractures in a touristic area were investigated and our concept for treatment presented. METHODS: Between 1.1.1999 and 30.04.2000 adult patients with proximal humeral fractures were included, the fractures classified (Codman/Neer and AO) and results determined after an average of 9 months. RESULTS: 62 adults were treated. 59 (95 %) had an accident during leisure time, mainly skiing accidents (52 %). 7 patients (11 %) had an associated luxation of the shoulder. 51 (82 %) were treated conservatively, 11 (18 %) operatively with a T-plate. The conservatively treated had to wear a Gilchrist-cast for an average of 29 (operatively 13) days, started passive movement after 20 (operatively 9) days, and active movement after 44 (operatively 45) days. The 32 employed (52 %) were not able to work for 46 days on average. Overall, 52 patients (84 %) were totally or mostly satisfied with the result. 5 among the 13 patients (38.5 %) with 3- or 4-part-fractures, and 4 among the 11 operated patients (36.4 %) were not satisfied with the result. CONCLUSIONS: Proximal humeral fractures are common skiing injuries, they need a long and intensive treatment and are economically expensive. The Codman/Neer and AO-classifications are equal. The results for simple, mainly conservatively treated fractures (Codman/Neer 1, 2A, 2-part) are good. Complex, mainly operatively treated fractures (Codman/Neer 3- and 4-part) have a much poorer prognosis. Diagnostically the computed tomography with 3-D reconstruction is recommended for a better representation of the fracture and a safer choice of the therapeutical strategy. PMID- 12632280 TI - [Primary hemiarthroplasty after complex fracture of the humeral head--functional late results]. AB - Treatment of complex fractures of the humeral head remains an obviously unsolved problem. Under consideration of the problems of head preserving methods, the development of the last years tended towards the hemiprosthetic replacement. As the discussion about the methods and implants continues to be controversial, long term results of Neer-II-replacement of humeral head in own trauma cases are presented here. PATIENTS AND METHODS: 46 patients were investigated, in whom a cemented Neer II hemiprosthesis was implanted between 1983 and 1996 due to complex fracture patterns. Median age was 60.2 years, 74 % of patients were female. 59 % had a luxation fracture type Neer VI, 22 % presented a concomitant nerval lesion. Follow up was performed 5 years on average after the operation. 38 patients were investigated clinically according the score of Constant and Murley, 36 had a x- ray control. Additionally an individual questionnaire was completed by 46 patients. RESULTS: Average Constant-score was 66.2 points, whereas in 76.3 % a satisfactory or better result was achieved. In 23.9 % the outcome was poor. In one patient (2.2 %) explantation of the hemiprosthesis and shoulder fusion due to pain was necessary. Conditions for a good result were a patient aged 50 years and younger (75.9 pts. versus 64.2 pts.), an early hemiprosthetic replacement within 4 weeks after the trauma (71.7 pts. versus 55.3 pts.) and a patient without an earlier operation and nerval lesion (76.8 pts. versus 53.9 pts.). No correlation was found between the date of the follow-up and the interval of time since the operation. CONCLUSION: After complex fractures of the humeral head satisfactory long-term results can be obtained by the cemented Neer II hemiprosthesis. Thus, implants of the newer generation can not yet whitness their superiority under consideration of functional aspects. Refixation and reintegration of both the tubercula after trauma still seems to be an unsolved problem and requires further development. PMID- 12632281 TI - [Intramedullary stabilisation of metastatic fractures of long bones]. AB - Surgical treatment of metastatic fractures of long bones is mostly a palliative one. Intramedullary stabilisation without resection of metastases follows the aim of the palliative therapy concept. PATIENTS AND METHODS: From 01.01.1995 to 30.08.2001 36 manifest and 4 impending long bone fractures were registered in 22 female and 16 male patients, with a mean age of 63 years. In addition, one revision was necessary due to persisting instability after humeral intramedullary bundle nailing. Pathological fractures were found in the humerus (n=11), femoral neck (n=6), per- and subtrochanteric region (n=12), femoral shaft (n=10) and tibia (n=2). Most of them were malignancies of the kidneys (n=13), bronchi (n=7) and breast (n=6). One patient with a solitary metastasis in the humerus received curative treatment by resection, open reduction and internal fixation. All other cases underwent palliative stabilisation. Metastatic lesions of the femoral neck were treated by resection and prosthetic replacement. The remaining fractures were stabilized intramedullarily without resection of metastases or use of bone cement. In 9 cases, a locking nail was implanted in the humerus, and one patient received humeral bundle nailing. In the lower extremities, we used a classic-nail or gamma-nail in 15 and a locking nail in 8 cases. Two moribund patients did not undergo surgery. RESULTS: No intraoperative complications occurred. 63 % of the patients with stabilisation of the leg were able to walk at the time of discharge. The mean survival time (Kaplan-Meier) of patients after palliative operation amounted to 161.5 days (95 % CI 92.7; 230.3). We found 6 postoperative complications: temporary palsy of the radial nerve (n=1), soft tissue infection (n=1), dislocation of the nail (n=2), loosening of the distal locking screw (n=1), proceeding osteolysis with high fracture risk (n=1). CONCLUSION: Intramedullary stabilisation without resection of metastases using locking nails meets the requirements of palliative therapy. This procedure is less invasive and allows early weight bearing. PMID- 12632282 TI - [Rupture of the gluteus medius tendon]. AB - AIM: Rupture of the gluteus medius tendon is a rare, even neglected condition. Typical findings include local tenderness and weakness of hip abduction. This pathologic condition is very seldom analysed in the literature, there is no report about the meaning in primary total hip replacement (THR). PATIENTS: Between 1/1/1995 and 12/31/2000 we found 13 cases (3,5 %) with a tear of the gluteus medius tendon in 372 primary THR using the lateral approach. In all cases the rupture was seen in the ventral and lateral part of the gluteus medius tendon. Furthermore, a valgus position of the proximal femur was detected. Transosseous refixation of the tendons was performed in all cases. 3 months after operation local tenderness near the greater trochanter disappeared and full strength of the pelvitrochanteric muscles was achieved in 11 out of 13 cases and in the remaining cases after one year. CONCLUSION: When a valgus position of the proximal femur in primary THR is found, one has to be aware of a simultaneous rupture of the gluteus medius tendon. This pathologic finding has to be ruled out preoperatively by means of ultrasound or MRI and an adequate approach to the hip joint has to be selected to examine the pelvitrochanteric muscles and to treat the rupture. PMID- 12632283 TI - [The significance of the pneumatic tourniquet technique for an innovative varicose vein surgery--methods, advantages, risks, danger]. AB - AIM: The systemic effects of the pneumatic tourniquet technique in varicose vein surgery were studied in order to evaluate possible risks and threats. METHODS: 155 patients were included to look at intraoperative changes of cardiovascular parameters and capillary blood and respiratory gases. RESULTS: Whereas the pulse rate showed only an insignificant rise of 3/min, the systolic and diastolic blood pressure increased significantly by 12 and 6 mm Hg resp., after inducing ischemia by roller cuff. After removal of the roller cuff blood pressure decreased by 14 and 9 mm Hg, resp., again. At releasing of the ischemia, pH values drop by 0.02, and the base excess by 0.3 mval/L with operation on one side and 0.5 mval/L in both sides operation. In comparison, the capillary pCO 2 rose by 3 mm Hg and the pCO 2 of the respiratory air by 1 mm Hg in one side operation and 3 mm Hg, resp. in operation of both sides. The pressure in the roller cuff was in the middle only 55 mm Hg above systemic pressure values. CONCLUSIONS: The systemic effects of the tourniquet technique have no clinical relevance. The use of the pneumatic tourniquet (Loefqvist method) is unrestrictedly recommended due to its advantages, such as no intraoperative blood loss, avoidance of post-op haematoma and pain and good immediate cosmetic results. PMID- 12632285 TI - [Lysis therapy of thrombosis of the superior mesenteric artery]. AB - INTRODUCTION: For surgeons the acute intestinal ischaemia is still a diagnostic and therapeutic challenge. If clinically suspected, the diagnostic procedures such as duplex sonography and arterial angiography should be carried out immediately. Although the diagnosis is often quickly clear, perioperative mortality rate remains high. We report the acute local thrombolytic therapy as an alternative treatment. CASE REPORT: A 80-year-old male patient was referred to our hospital with a complete occlusion of the superior mesenteric artery. Duplex sonography and the arterial angiogram confirmed the clinical diagnosis. Because the patient was assessed to be at high risk we decided to avoid an operation and local thrombolytic therapy using rt-PA and urokinase was carried out. RESULT: The local thrombolytic therapy was successful and led to a complete restoration of the arterial flow within the superior mesenteric artery. The clinical symptoms subsided and no complications were observed. CONCLUSION: Local thrombolytic therapy appears to be a suitable therapeutic option in patients suffering of mesenteric arterial occlusion. Although the duplex sonography often confirms the diagnosis with high accuracy the angiography remains the diagnostic gold standard. PMID- 12632284 TI - [Internal carotid artery dissection at the carotid canal after blunt trauma: case report of endovascular repair and literature review]. AB - Symptomatic dissecting aneurysms of the extracranial internal carotid artery after blunt trauma are not frequent. Medical treatment of these lesions can be ineffective and surgical options present high morbidity. We describe a case successfully treated with endovascular therapy, using a self-expanding stent. The advantages of the endovascular treatment are outlined, the literature on carotid artery injuries by blunt trauma and management strategies are reviewed. PMID- 12632286 TI - [Experimental investigations onto the biocompatibility of coated vascular grafts as a function of the primary disease in various patients groups]. AB - INTRODUCTION: This work presents initial experimental results obtained with co incubation of human neutrophilic granulocytes from patients with various diseases (5 patients each group) and collagen- or silver-coated polyester grafts (25 patients each graft). METHODS: After two-hour co-cultivation of the graft with human granulocytes of different patients (normal controls; peripheral occlusive vascular disease; diabetes mellitus; carcinoma/chemotherapy; sepsis), the following parameters were determined by flow cytometry: CD11b, CD62L, fMLP, CXCR2. RESULTS: The change in the receptor expression was taken as a measure of responsiveness, with statistically significant differences seen within and between the groups. For the silver-coated graft, such differences were mainly noted for the fMLP receptor (p=0.01). For the two-hour incubation with collagen coating, the receptors CD62L and CXCR2 were indicative of differences between the various diseases (CD62L p=0.01; CXCR2 p=0.01). Comparison between the grafts revealed statistically significant differences for the CXCR2 and the fMLP receptors (CXCR2 p=0.00; fMLP p=0.03). CONCLUSIONS: The differences noted between the grafts and between the patients groups are suggestive of an altered responsiveness of the granulocytes to the grafts. This enables new aspects with respect to the genesis of accompanying clinical symptoms. PMID- 12632291 TI - [44th congress of the German Society of Pneumology 26. 3. - 29. 3. 2003 in Munich]. PMID- 12632292 TI - [Miflonide/Foradil via Aerolizer compared with other anti-inflammatory and anti obstructive therapeutic regimens]. AB - Standard therapy of asthma consists of combined, antiinflammatory (steroids) and anti-obstructive (long acting/short acting beta-agonist) treatment via inhalation using two separate or a single inhalation device. Here, we report the results of using Miflonide (Budesonide 400 - 800 microg per day) and Foradil (Formoterol 24 48 microg per day) in cases of moderate and severe asthma previously treated insufficiently with another combination therapy. 80 patients with asthma previously treated insufficiently with any other combination therapy of steroid and long acting beta-agonist were included. Instead of their previous therapy all were switched to therapy with Miflonide and Foradil for eight weeks (two office visits at 4 and 8 weeks). Lung function (peak flow [l/min], FEV1 [I], FVC [I], R(tot) [kPa*s/l]) was performed at every visit. Doctors' and patients' estimation of disease severity, physical examination, drug related side-effects and the use of short acting beta-agonist aerosols were registered. Lung function parameters improved significantly compared to the run in phase prior to the change in medication (peak flow: + 18.4 %, FEV 1 : + 10.7 %, FVC: + 6.8 %, R(tot) : -18.0 %). Use of additional short acting inhalative beta-agonists was reduced. Subjectively, patients judged their general condition as improved, effectiveness as greater compared to previous medication and side effects as tolerable. The use of a combination of Miflonide/Foradil lead to an improvement in subjective and objective parameters in asthma patients that had previously been treated with a variety of other antiinflammatory and anti-obstructive therapy regimens. Reasons for this observation are beside change in medication, patients training in asthma therapy, change of application system, and increase of patients compliance. PMID- 12632293 TI - [The yellow nail syndrome]. AB - A case of a 54-year-old woman exhibiting the yellow nail syndrome with exudative pleural effusions, lymphoedema, pericardial effusion, rhinosinusitis and bronchiectasis is described. This case illustrates the rather benign course of the syndrome over ten years. A review of the literature is provided. The etiology of the syndrome is obscure, while the pathogenesis seems to involve impaired lymphatic drainage. The entity should be considered in the differential diagnosis of bilateral pleural effusion. It is likely that the condition is not as rare as it appears from the number of published reports, and a greater awareness of its various forms will probably lead to more frequent diagnosis. PMID- 12632294 TI - [Occupational aziridine asthma in an engineer at a printing office]. AB - We report on a case of an allergic bronchial asthma of an engineer in a printing office caused by polyfunctional aziridines. The causal relationship between exposure and symptoms could be demonstrated with a specific bronchoprovocation challenge testing. The consequences of the disease are discussed with respect to medical and legal aspects of the system of social security in Germany. PMID- 12632295 TI - [Difficulties in controlled inhalation of alpha 1-protease inhibitor]. AB - In this paper a number of studies will be summarized which were designed to improve the inhalation of alpha 1 -protease inhibitor in patients with alpha 1 protease inhibitor deficiency. A pilot study has shown that the high inter individual variability of drug deposition in the lungs is due to heterogeneous breathing patterns of the patients. Controlling the breathing pattern led to a significantly decreased variability. Then it was studied which particle size and breathing pattern resulted in highest peripheral lung deposition in patients with emphysema. It was found that for 3 - 4 microm particles and slow inhalation flow rate the peripheral deposition increases with increasing inhalation volume. After the development of an inhalation device which allows to perform controlled inhalations in clinical practice it was shown that this device, in combination with a breathing pattern individually normalized to the patients lung function, allows to deposit nearly 60 % of the drug into the patients lung periphery. PMID- 12632296 TI - [Pulmonary Langerhans cell histiocytosis]. PMID- 12632298 TI - [Aerosols in medicine and veterinary medicine]. PMID- 12632301 TI - Anterolateral thigh flap for breast reconstruction: review of the literature and case reports. AB - More women than ever before are undergoing mastectomies secondary to increased awareness and screening. This has also caused a corresponding increase in the number of breast reconstructions requested each year. The demand for improved results has fueled recent advances in new techniques. Aside from implant reconstruction, the methods now being employed are related to autogenous donations and reconstruction. Currently, the most commonly used techniques for autogenous breast reconstruction are the DIEP (deep inferior epigastric perforator) and TRAM (transverse rectus abdominis myocutaneous) flaps from the lower abdomen. The anterolateral thigh flap is a type of perforator flap usually described for use in head and neck reconstruction. The authors have discovered this flap's utility as an alternative in autogenous breast reconstruction when the abdomen is not available as a donor site. A review of the literature reveals a dearth of experience in using the anterolateral thigh flap for breast reconstruction. The article reviews the literature with regard to current uses of the anterolateral thigh flap, and then reports three case studies which highlight the thigh flap as an excellent alternative for breast reconstruction in selected patients. PMID- 12632302 TI - Treatment of meralgia paresthetica with a deep inferior epigastric perforator adiposal flap: case report. AB - A case of a 40-year-old female with meralgia paresthetica after malignant tumor resection in the right inguinal region is reported. Traditionally, meralgia paresthetica is treated with neurotransection or neurolysis. The therapeutic strategy using neurolysis, and the use of a deep inferior epigastric perforator adiposal flap wrapping as a prophylactic procedure against reentrapment is discussed. PMID- 12632303 TI - Reconstruction of a complex hemifacial deformity with multiple simultaneous free flap transfers: case report. AB - Severe disfigurement, facial paralysis, abnormal continuity of oral and nasal passages, and velopharyngeal incompetence resulted, following maxillectomy and cranial-base resection for a radiation-induced sarcoma. Oral lining, bone support, facial muscle, and outer skin were provided with a single-staged transfer of radial forearm, scapular bone, scapular skin, and serratus anterior muscle with anastamoses to the contralateral neck. This combination of flaps may be considered in complex facial deformities, and may obviate the need for staged prefabrication. PMID- 12632304 TI - Repair of avulsion injury of the whole hand with single-stage transfer of five combined tissues: case report. AB - The authors present a 32-year-old man with an entire left hand avulsion injury. This was repaired in one stage with five free-tissue transfers, a combination of a left hallux wrap-around flap for thumb reconstruction, bilateral second toes for middle and ring finger reconstruction, and bilateral femoral anterolateral flaps for coverage of the injured hand at the palmar and dorsal aspects. All the transfers survived uneventfully. Due to enlargement of the flaps and narrowness of the first web space, three further surgical procedures were subsequently carried out. Over a 2-year follow-up, a satisfactory appearance of the reconstructed hand and functional restoration were obtained. PMID- 12632306 TI - Successful replantation of an amputated leg with severe crush and avulsion injury in an elderly patient: case report. AB - An amputated leg with severe crush and avulsion injury was replanted in a 57-year old man. Since the skin and soft tissue around the ankle became necrotic 11 days postoperatively, a flow-through forearm free flap was transferred into the defect, preserving the blood circulation to the replanted leg. Seven weeks after the replantation, sural nerve cable grafts were interposed between both amputated edges of the tibial nerve. As the function of the ankle joint did not recover, arthrodesis of the ankle joint was performed 7 months after the replantaion. As a result, the patient could walk 1 year postoperatively. PMID- 12632307 TI - Second free jejunal transfer in complicated pharyngoesophageal reconstructions. AB - The authors report two cases of pharyngoesophageal reconstruction using free jejunum transfer, which required additional microsurgical reconstruction, due to total or partial loss of the free jejunal graft, despite successful microvascular anastomoses. Replacement and additional transfer of the second free jejunum segment were successfully carried out, with minimal risk and low morbidity. The second free jejunal transfer is one of the reliable reconstructive choices in the management of complicated pharyngoesophageal reconstruction with free jejunum. PMID- 12632308 TI - Vascularized second metacarpal-base bone graft in scaphoid non-union by the palmar approach. AB - Vascularized second metacarpal-base bone grafting from the palmar side for scaphoid non-union was performed in seven patients (six males and one female, with a mean age of 26 years). In all patients, bone union was obtained at a mean of 10.2 weeks. In six of the seven patients, preoperative dorsal intercalated segment instability (DISI) was corrected. The results were excellent in three patients, good in two patients, and fair in two patients, showing that more than 80 percent of the patients had satisfactory results. In this procedure, the second dorsal metacarpal artery, with a large vascular diameter, is used as the pedicle, and the second metacarpal base, which is abundant in cancellous bone, is elevated on the dorsal side and grafted on the open-wedged scaphoid non-union on the palmar side. This procedure is reliable and useful, particularly for scaphoid non-union with DISI. PMID- 12632309 TI - Early compensatory sensory re-education. AB - After a neurorrhaphy, there will be a distal disconnection between the cortex and skin receptors, along with interruption of sensibility information. This report demonstrates the efficacy of a new sensory re-education program for achieving optimal sensation in a relatively short time. Between 1999 and 2001, in the authors' Hand Rehabilitation Department, 11 patients with previous neurorrhaphy were subjected to a program of early "compensatory sensory re-education." Lesions were caused by clean cut. There were 13 primary digital nerve procedures, 12 at the distal palmar MP level, and one at the radial dorsal branch of the index (just after emerging from the common digital nerve). The technique of compensatory sensory re-education was based on a previous, but modified, sensory re-education method. In order to evaluate the results in the compensatory sensory re-education series described, additional tests for evaluation of achieved functional sensibility were used. The authors' best results were achieved in a maximum of 8 weeks (4-8 weeks), much less time than with the original method (1-2 years). Using the British classification, it was possible to compare the achieved levels of sensibility and the time required for optimal results. The different methods of sensibility re-education may be similar, but with the authors' compensatory sensory re-education method, substantial time is saved. PMID- 12632311 TI - Effects of delaying FK506 administration on neuroregeneration in a rodent model. AB - FK506 is an immunosuppressant drug that has been shown experimentally to stimulate nerve growth and speed functional recovery, when administered immediately after peripheral nerve injury. However, the clinical scenario of a peripheral nerve injury is often associated with either a delayed diagnosis or reconstruction. The purpose of this study was to determine the efficacy of FK506 on neuroregeneration with delayed administration. Thirty-two Lewis rats underwent tibial nerve transection with immediate repair. Animals were left untreated, or were treated with daily injections of FK506 (2 mg/kg), started on the day of surgery, postoperative day 3, or postoperative day 5. Animals underwent walking track analysis to assess functional nerve recovery. Nerves were harvested for histomorphometric analysis on postoperative days 21, 28, and 42. Histomorphometry demonstrated that all treatment groups, regardless of the time of drug initiation, demonstrated evidence of enhanced neuroregeneration, compared to the untreated group. Histomorphometric data from groups harvested on day 21 demonstrated a statistically significant improvement in neuroregeneration in the immediate and 3-day delay groups. Therefore, the beneficial effects of FK506 on neuroregeneration are not restricted to immediate administration, but these effects significantly diminish when FK506 is administered 3 days after nerve injury. PMID- 12632312 TI - Schwann-cell proliferation in muscle-vein combined conduits for bridging rat sciatic nerve defects. AB - Among the various grafting procedures that have been studied as alternatives to traditional fresh nerve autografts for the repair of severed peripheral nerves, muscle-vein-combined graft conduits have recently been devised and successfully employed. In the present study, the early presence, origin, and proliferation activity of Schwann cells (SCs) along this particular type of biological graft conduit have been investigated, using antibodies directed against glial fibrillar acid protein (GFAP), a protein that is specifically expressed in glial cells, and proliferating cell nuclear antigen (PCNA), a protein that is expressed by cells during DNA synthesis. Results showed that the muscle-vein-combined graft was progressively invaded by a number of GFAP-immunopositive SCs, many of which were also found to be immunopositive for PCNA, thus demonstrating that their proliferation continues to occur inside the graft. Among the molecules that could be involved in the stimulation of Schwann-cell proliferation is neuregulin-1 (NRG 1) that mediates its effects by binding to the ErbB receptor tyrosine kinase family. In the present study, the authors report on the RT-PCR analysis for NRG-1 and ErbB3 mRNAs, showing an overall increase in the content of these transcripts inside the muscle-vein-combined graft. These results suggest that the muscle-vein combined graft conduit constitutes an environment favorable to potentiate Schwann cell proliferation during the early regeneration phases. PMID- 12632315 TI - [Heroin-assisted treatment of opioid addicts in Switzerland from 1994-2001- utilisation and characteristics of admissions and discharges]. AB - BACKGROUND OBJECTIVES: Since the beginning of the 1990 s the number of treated opioid addicts has markedly increased in Switzerland. This study examines the cause of health service utilisation of one specific type of treatment--heroin assisted maintenance--the length of stay in this kind of treatment, and characteristics of admissions and discharges. METHODS AND SAMPLE: Data on all admissions into and discharges from heroin-assisted treatment were collected from the start of this treatment modality on January 1, 1994 to December 31, 2001. These data were used to calculate length of stay, and number of patients at the end of each calendar year. Data were analysed according to sex and age and the Kaplan-Meier survival statistics were calculated. Finally, the discharges from 1994 to 1998 were compared to the discharges of 1999 up to December 2001. RESULTS: Since the introduction of heroin-assisted treatment, the number of patients in this form of treatment has markedly increased. Simultaneously, the average age of the patients entering treatment increased, whereas the proportion of women decreased continually. Compared to the earlier phase (1994-1998), in the last three years fewer patients terminated treatment at an early stage (within the first 4 months). CONCLUSIONS: Substitution programmes are the most common form of treatment for opioid dependence in Switzerland. The increase in the average age of patients in this treatment modality as well as in the abstinence oriented treatment at admission may be interpreted favourably as an indicator that fewer less young people are opioid dependent than at the beginning of the 1990 s. The decrease in the proportion of females may be an indicator that females were overrepresented in heroin-assisted treatment at the beginning of this treatment modality and that the proportion gradually became more similar to the proportion in the opioid-dependent population as a whole. The decrease of early treatment terminations can be seen as positive, since with longer length of stay the probability of changing into abstinence-oriented treatment is becoming more probable. PMID- 12632314 TI - Enhanced peripheral nerve regeneration elicited by cell-mediated events delivered via a bioresorbable PLGA guide. AB - Using an established rat peripheral-nerve regeneration model, the authors have demonstrated enhancement of regeneration following subcutaneous priming of bioresorbable poly(lactic-co-glycolic)acid (PLGA) guides in vivo. Four weeks after nerve reconstruction, regeneration of the peripheral nerve through the cell infiltrated guides displayed a significant increase in the total axon number and myelination status recorded in primed over unprimed guides, demonstrating the importance of cell-mediated events in the regeneration process. To define the different components enhancing nerve regeneration in this model, they have focused on identifying factors capable of eliciting Schwann-cell migration, since this has been identified as an early and necessary event in nerve regeneration. Using an in vitro migration assay, screening of a limited number of cellular and extracellular factors has demonstrated differential promotion of Schwann-cell migration. Of interest, combining fibronectin and bFGF resulted in a two-fold enhancement in Schwann-cell migration over that recorded with either alone. These results describe a rapid screening process for identifying various molecules and combinations thereof, with potential involvement in Schwann-cell migration. Coupling these findings to the use of the PLGA guide as an in vivo delivery system provides a rationale for the selection of exogenous factors to test for the enhancement of peripheral-nerve regeneration. PMID- 12632316 TI - [Inpatient withdrawal treatment of patients suffering from alcohol addiction in Berlin (1993-1999)]. AB - On account of the Hospital Statistics Regulation from 1990 all hospitals have to run a diagnosis-related and regionally structured hospital documentation, beginning with the year 1993. In the State of Berlin, this routine documentation from 74 hospitals of Berlin is centralised in the Statistics State Office. We obtained the data records with the main diagnosis alcoholism (303 ICD 9) for the years 1993 to 1999 for evaluation. The treatment frequency is male dominated with average yearly 400 vs. 100 female cases per 100 000 inhabitants. In East Berlin men reached due to continuous increase of cases for the first time in 1998 higher values in comparison with West Berlin. The treatment frequency of female West Berlin patients is in the period under review continually about twice as high than that of female East Berlin patients. The age peak for both sexes lies between 40 and < 50 years. The patients were especially treated in the wards of "psychiatry" (ca. 40 %), of "addiction diseases" (ca. 30 %) and "internal medicine" (ca. 23 %). The average hospital retention period was reduced from 22 (1993) to 12 days (1999). PMID- 12632317 TI - [Structures of co-operation between an HIV-outpatient clinic and private care physicians]. AB - OBJECTIVES OF THE STUDY: This qualitative study looks at structures of co operation between the two major HIV-care settings in the region of Zurich (Switzerland), i.e. a HIV-specialised outpatient clinic at the University hospital and private practitioners. Hypothesizing that good co-operation between the institutions increases the quality and effectiveness of care for patients living with HIV and treated with highly active antiretroviral therapy (HAART), the structures of co-operation are investigated. METHODS: Qualitative data were collected by means of three focus-group discussions with physicians from both settings. By means of a structured interview guide, different aspects of co operation were assessed (e. g. current status, deficits, degree of satisfaction). Contents analysis of the data revealed consensus versus dissonance pertaining to different areas of co-operation. RESULTS: Results show that co-operation has been implemented effectively on three different planes (case-related, knowledge related, training-related) and generally has been perceived by the participants to function well. Institutions have different emphasis on care (specialists vs. generalists). They formulate different concepts of co-operation, which result in diverse degrees of satisfaction. CONCLUSIONS: On the basis of the results obtained, suggestions for improved co-operation as recommended by the participants are discussed, which could further increase the quality of HIV related care. PMID- 12632319 TI - [The "Spandau Public Health" test--description of study]. AB - In 2002 the "Spandauer Gesundheitstest" ("Spandau Public Health" Test), a prospective cohort study of the Robert Koch-Institute in Berlin has been successfully going on for two decades. Ten waves of follow-up of this study are now available, in which approximately 7 000 adults were interviewed and examined. Approximately 770 participants have taken part in all ten follow-up waves. The "Spandauer Gesundheitstest" is carried out to provide information on changes in health state at the individual level over time. The main epidemiological aim of the study is to follow up the development and course of chronic disease together with changes in health care needs and utilisation. A questionnaire, blood and urine examinations and a medical interview are parts of the study design in every wave. Since the beginning of this study more than 900 participants died. A mortality follow-up is carried out regularly to investigate the cause of death of the participants. The data are available for analysis. PMID- 12632318 TI - [Comparative study on the dental health of German and immigrant 8- to 10-years olds in the Westphalian Ennepe-Ruhr district]. AB - Aim of this 2-year follow-up study was to assess the dental health status and the caries pattern of first permanent molars in German and immigrant children. 300 German and 69 immigrant children living in the Westphalian Ennepe-Ruhr district were involved in this study. The mean age of the population was 8.0 years at the baseline (1997) and 9.8 years at the second examination. The surface-related caries status according the WHO criteria (1997) was completed by recording non cavitated lesions and the number of sealants in permanent molars. 13 % (1997) and 26 % (1999) of the 8-year old immigrant children had a caries-free mixed dentition; in Germans were recorded 40 % (1997) and 42 % (1999). The caries prevalence in immigrant children (1997: 0.9 DMFS/1999: 1.5 DMFS) was significantly higher compared to the German children (1997: 0.5 DMFS/1999: 0.8 DMFS). Moreover, a significantly lower utilisation of fissure sealants was registered in immigrant children (1997: 0.4/1999: 0.7) compared to their German classmates (1997: 0.5/1999: 1.4). Immigrant children have to be characterized as high caries-risk population. Risk-orientated public dental health programmes involving sealant application seems to be necessary for socio-economically underprivileged children. PMID- 12632320 TI - [Medical care research based on family doctor routine data--are interface communicated treatment data feasible?]. AB - AIMS OF THE STUDY: As part of an ongoing project on the utilisation of generic drugs in general practice we aimed at determining whether the transfer of prescriptions and patient characteristics from doctors' computerized medical records via the BDT (Behandlungsdatentrager) interface was feasible, and whether these data are suitable for research in pharmacoepidemiology. METHODS: All 1,395 general practitioners from 6 regions in Germany were invited to participate in the 'generics project'; 232 (17 %) agreed. The 17 software companies whose systems were used by the participating practices were asked to grant access to the BDT interface. For a prescription survey, doctors were supposed to export BDT files from two 3-month periods each in 2000 and 2001. Data were anonymised and relevant information extracted with a special programme. RESULTS: So far, BDT data are available from 79 practices. They are suitable for practice- and patient related prescribing analyses. By filter modifications, additional information (such as diagnoses, referrals, clinical findings or accounting codes) can be obtained. The procedure was well accepted if doctors and practice staff were assisted by computer experts. Some difficulties, however, were encountered in obtaining access to the BDT-interface from the software companies. Lack of standardisation of the BDT interface required additional conditioning of the data. CONCLUSION: The BDT interface offers an opportunity to export computerised patient records without the requirement of additional documentation. If routine data are more readily available for health services research, a standardised data structure and open access must be assured e. g. by centralised certification via the Federal statutory health organisation. PMID- 12632322 TI - [Health impact assessment of building and investment projects]. AB - For regional planning and approval procedures for building projects of a certain order of magnitude and power rating according to the German Federal Act on the Prevention of Emissions with Integrated Environmental Impact Assessment (EIA), the German public health departments, acting as public authorities, increasingly perform health impact assessments (HIA). The amended Act on Environmental Impact Assessment, the Decree on industrial plants which require approval (4th Federal Decree on Emission Prevention) and the Health Service Acts of the Federal States of Germany form the legal basis for the assessment of health issues with regard to approval procedures for building and investment projects. In the framework of the "Action Programme for the Environment and Health", the present article aims at making this process binding and to ensure responsibility and general involvement of the Public Health departments in all German Federal States. Future criteria, basic principles and procedures for single-case testing as well as assessment standards should meet these requirements. The Federal Ministry for the Environment and the Federal Ministry for Health should agree on Health Impact Assessment (HIA ) as well as on the relaxant stipulations in their procedures and general administrative regulations for implementing the Environmental Impact Assessment Act (EIA). Current EIA procedures focus on urban development and road construction, industrial investment projects, intensive animal husbandry plants, waste incineration plants, and wind energy farms. This paper illustrates examples meeting with varying degrees of public acceptance. However, being involved in the regional planning procedure for the project "Extension of the federal motorway A 14 from Magdeburg to Schwerin", the Public Health Service also shares global responsibility for health and climate protection. Demands for shortest routing conflict with objectives of environmental protection which should be given long term consideration. Assessing the direct impact of projects on human beings should be rank first in the list of priorities. The Hygiene Institute supports the efforts of the Public Health departments by providing professional consultant services to ensure consistency in the application of procedures. PMID- 12632321 TI - [Female incontinence: problems with assessment of prevalence and severity]. AB - Incontinence raises a problem of increasing relevance. Estimates of prevalence and severity are presented in a wide range. Valid assessment instruments are sparse or based on varying definitions. Therefore, existing data of incontinence rates are likely to be overestimated. To exemplify this problem, we reviewed epidemiological articles on female incontinence. PMID- 12632323 TI - [Demands for an internationalisation of German research as illustrated by the example of the journal "Das Gesundheitswesen". A contribution to the quality assurance of applied research]. AB - OBJECTIVES: Basing on the recent discussion on internationalisation of German research publications, the most recent volume of "Das Gesundheitswesen" was bibliometrically analysed. METHODS: 125 articles were compared with respect to citing sources, the rate of self-citations, the number of times English or German journals or books were cited in the year 2001, the acceptance rate of the international state of research and of its leading representatives. These are indicators of scientific quality. RESULTS: The interdisciplinary approach by "Das Gesundheitswesen" has proven successful in view of certain indicators of scientific quality. CONCLUSION: Related to scientific quality standards "Das Gesundheitswesen" has achieved a good impact in the scientific community but some details require to be changed. PMID- 12632324 TI - [The receipt to be given to the patient: a discussion]. PMID- 12632325 TI - Rapid direct sequence analysis of the dystrophin gene. AB - Mutations in the dystrophin gene result in both Duchenne and Becker muscular dystrophy (DMD and BMD), as well as X-linked dilated cardiomyopathy. Mutational analysis is complicated by the large size of the gene, which consists of 79 exons and 8 promoters spread over 2.2 million base pairs of genomic DNA. Deletions of one or more exons account for 55%-65% of cases of DMD and BMD, and a multiplex polymerase chain reaction method-currently the most widely available method of mutational analysis-detects approximately 98% of deletions. Detection of point mutations and small subexonic rearrangements has remained challenging. We report the development of a method that allows direct sequence analysis of the dystrophin gene in a rapid, accurate, and economical fashion. This same method, termed "SCAIP" (single condition amplification/internal primer) sequencing, is applicable to other genes and should allow the development of widely available assays for any number of large, multiexon genes. PMID- 12632326 TI - Missense mutations in CRELD1 are associated with cardiac atrioventricular septal defects. AB - Atrioventricular septal defects (AVSD) are common cardiovascular malformations, occurring in 3.5/10,000 births. Although frequently associated with trisomy 21, autosomal dominant AVSD has also been described. Recently we identified and characterized the cell adhesion molecule CRELD1 (previously known as "cirrin") as a candidate gene for the AVSD2 locus mapping to chromosome 3p25. Analysis of the CRELD1 gene from individuals with non-trisomy 21-associated AVSD identified heterozygous missense mutations in nearly 6% of this population, including mutations in isolated AVSD and AVSD associated with heterotaxy syndrome. CRELD1 is the first human gene to be implicated in the pathogenesis of isolated AVSD and AVSD in the context of heterotaxy, which provides an important step in unraveling the pathogenesis of AVSD. PMID- 12632333 TI - Managing cholesterol with plant stanol esters. PMID- 12632327 TI - Homozygous mutations in IHH cause acrocapitofemoral dysplasia, an autosomal recessive disorder with cone-shaped epiphyses in hands and hips. AB - Acrocapitofemoral dysplasia is a recently delineated autosomal recessive skeletal dysplasia, characterized clinically by short stature with short limbs and radiographically by cone-shaped epiphyses, mainly in hands and hips. Genomewide homozygosity mapping in two consanguineous families linked the locus to 2q35-q36 with a maximum two-point LOD score of 8.02 at marker D2S2248. Two recombination events defined the minimal critical region between markers D2S2248 and D2S2151 (3.74 cM). Using a candidate-gene approach, we identified two missense mutations in the amino-terminal signaling domain of the gene encoding Indian hedgehog (IHH). Both affected individuals of family 1 are homozygous for a 137C-->T transition (P46L), and the three patients in family 2 are homozygous for a 569T- >C transition (V190A). The two mutant amino acids are strongly conserved and predicted to be located outside the region where brachydactyly type A-1 mutations are clustered. PMID- 12632328 TI - ATLAS: a system to selectively identify human-specific L1 insertions. AB - Retrotransposition of L1 LINEs (long interspersed elements) continues to sculpt the human genome. However, because recent insertions are dimorphic, they are not fully represented in sequence databases. Here, we have developed a system, termed "ATLAS" (amplification typing of L1 active subfamilies), that enables the selective amplification and display of DNA fragments containing the termini of human-specific L1s and their respective flanking sequences. We demonstrate that ATLAS is robust and that the resultant display patterns are highly reproducible, segregate in Centre d'Etude du Polymorphisme Humain pedigrees, and provide an individual-specific fingerprint. ATLAS also allows the identification of L1s that are absent from current genome databases, and we show that some of these L1s can retrotranspose at high frequencies in cultured human cells. Finally, we demonstrate that ATLAS also can identify single-nucleotide polymorphisms within a subset of older, primate-specific L1s. Thus, ATLAS provides a simple, high throughput means to assess genetic variation associated with L1 retrotransposons. PMID- 12632334 TI - Presidential address. Da Vinci and the Penrose drain. PMID- 12632335 TI - Fetal surgery: trials, tribulations, and turf. PMID- 12632336 TI - A prospective, randomized pilot trial of perfluorocarbon-induced lung growth in newborns with congenital diaphragmatic hernia. AB - BACKGROUND/PURPOSE: Initial laboratory and clinical data suggest that partial liquid ventilation (PLV) can enhance pulmonary function and that lung growth can be induced via distension of the newborn lung using perfluorocarbon in patients with congenital diaphragmatic hernia (CDH). The authors, therefore, performed a prospective, randomized pilot study evaluating PLV and perfluorocarbon-induced lung growth (PILG) in newborns with CDH on extracorporeal life support (ECLS) at 6 medical centers. METHODS: Patients were selected randomly using a permuted block design to PLV/PILG (n = 8) or conventional mechanical ventilation (CMV/control, n = 5). Patients in the PILG group received daily doses which filled the lungs with perflubron for up to 7 days and were placed on continuous positive airway pressure of 5 to 8 cm H2O. CMV patients were treated with standard mechanical ventilation while on extracorporeal membrane oxygenation (ECMO). RESULTS: A total of 13 patients were evaluated in this study. All 3 patients enrolled without being on ECLS rapidly transitioned to ECLS. The study, therefore, effectively evaluated PILG (n = 8) versus standard ventilation (control, n = 5) on ECLS. Mean (+/- SE) gestational age was 37 +/- 1 weeks and weight was 3.1 +/- 0.1 kg. Time on ECMO was 9.8 +/- 2.3 days in the PILG and 14.5 +/- 3.5 days (P =.58) in the control group. Survival rate in the PILG group was 6 of 8 (75%), whereas survival rate was 2 of 5 (40%) in the control group (P =.50). The number of days free from the ventilator in the first 28 days (VFD) was 6.3 +/ 3.3 days with PILG and 4.6 +/- 4.6 days with control (P =.9). Causes of death in the PILG group included sepsis and renal failure in one patient and pulmonary hypertension in the other. There were no safety issues, and the deaths in the PILG group did not appear to be related to the administration of perflubron. CONCLUSIONS: These data show that PILG can be performed safely. The survival rate, VFD, and time on ECMO data, although not conclusive, are encouraging and indicate the need for a definitive trial of this novel intervention in these neonates with high mortality. PMID- 12632337 TI - Impact of ECMO on neonatal mortality in Michigan (1980-1999). AB - PURPOSE: Although extracorporeal membrane oxygenation (ECMO) is a potentially lifesaving intervention, the effect of ECMO on neonatal mortality has never been evaluated. In this study, we examined the relationship between increased ECMO utilization and its effect on the neonatal mortality rate in the state of Michigan. METHODS: Neonatal mortality data were obtained from the Michigan State Department of Community Health. Data included total annual live births, total neonatal deaths, and deaths from respiratory causes in neonates > or =35 weeks' gestational age (ie, potential ECMO candidates). Pooled ECMO patient data from Michigan's 3 ECMO centers were obtained from the Extracorporeal Life Support Organization (ELSO) Registry. Associations between ECMO volume and neonatal mortality rates were assessed using simple linear regression. A scatterplot of ECMO volume and mortality rates was created, and the resulting fitted regression lines were superimposed on the plots. Statistical significance of the associations (ie, difference in slope of the regression line from zero) were based on a standard 2-sided Wald test for the regression slope parameter. RESULTS: From 1980 through 1999, 1,061 neonates were treated with ECMO in Michigan, and 875 (82.5%) survived. When annual neonatal mortality rate (from all causes) and the rate from respiratory causes versus ECMO volume (cases per year) are superimposed with fitted regression lines, both are significantly different from zero (P =.041 and P =.002, respectively). The model predicts that for every 100 neonates treated with ECMO in Michigan, 38 lives are saved. The model also can be used to predict the annual neonatal mortality rate in Michigan if ECMO had not been utilized. CONCLUSIONS: A strong association exists between ECMO volume and observed reductions in neonatal mortality seen in Michigan over the last 2 decades. This is the first study to show an association between ECMO and neonatal mortality rate at the population level. PMID- 12632338 TI - Reversed latissimus dorsi muscle flap for repair of recurrent congenital diaphragmatic hernia. AB - BACKGROUND/PURPOSE: Neonates with large congenital diaphragmatic hernias (CDH) require prosthetic patch closure of the defect because of the paucity of native diaphragmatic tissue. As the child grows, patch separation can occur necessitating reoperation. Use of vascularized autologous tissue may decrease the incidence of reherniation as tissue incorporation and growth may be improved. The authors report our early experience using a local muscle advancement flap with microneural anastomosis for those children in whom reherniation develops after prosthetic patch placement. METHODS: Seven patients with CDH (6 left and 1 right) whose synthetic diaphragmatic patch separated from the chest wall resulting in a clinically significant recurrent hernia were followed up with prospectively. After dissecting the ipsilateral latissimus dorsi off the chest wall and dividing the thoracodorsal neurovascular bundle (based on its lumbar blood supply), the synthetic patch was removed via an eighth intercostal incision. The muscle flap was placed into the hemithorax through the bed of the tenth rib and sutured in place over a Vicryl mesh scaffold. The thoracodorsal nerve was anastomosed to the phrenic nerve. Functional analysis of the flap was performed in 4 patients. RESULTS: Age at placement of the muscle graft ranged from 2 months to 48 months (median, 24 months). There has been no evidence of reherniation after placement of the muscle graft. Long-term outcome and functional analysis of the flap was available in 4 patients (mean, 19 months). Two infants had fluoroscopic and sonographic evidence of nonparadoxical neodiaphragmatic motion. In one of these, electromyographic evidence of function was documented with a phrenic nerve conduction velocity of 22 meters per second. The third infant showed no evidence of neodiaphragmatic motion, and the fourth infant had paradoxical motion. CONCLUSIONS: This is the first direct documentation of phrenic nerve function in an infant with CDH. An innervated reversed latissimus dorsi (RLD) flap reconstruction for recurrent CDH provides an alternative to prosthetic patch repair. This technique offers the advantages of autologous vascularized tissue with potential phrenic nerve innervation and physiologic neodiaphragmatic motion. PMID- 12632339 TI - Synergistic interleukin-18 and low-dose interleukin-2 promote regression of established murine neuroblastoma in vivo. AB - BACKGROUND/PURPOSE: Severe systemic toxicities have limited the clinical applications of the potent cytokine, interleukin-2 (IL-2). Recent studies have shown that IL-18 synergizes with IL-2 to enhance cytolytic activity in vitro. Combination therapy allows for IL-2 dose reduction, thus, limiting its toxicity while augmenting natural killer cell activity. The authors hypothesize that IL-18 plus low-dose IL-2 may induce a potent and sustained antitumor response in vivo providing effective immunotherapy for neuroblastoma. METHODS: Four groups of A/J mice (n = 28) were inoculated subcutaneously in the right flank with 1 x 10(6) murine neuroblastoma cells (TBJ). On day 7, 5 consecutive daily peritumoral injections were performed with saline (control), human rIL-2 (30,000 IU), murine IL-18 (1 microg), or IL-2 plus IL-18. Tumor growth was monitored, and animals with tumor progression were killed on day 21. Seven weeks after the initial treatment, animals with rejected tumors were rechallenged with 5 x 10(6) cells in the opposite flank. Quantitative data were analyzed by Student's t test. RESULTS: Rapid tumor growth and death was noted in all control animals by 21 days. Complete tumor eradication was seen in 28% of mice treated with IL-2 (P =.03), 42% of mice treated with IL-18 (P <.05), and 57% of mice treated with of IL-2 plus IL-18 (P <.05). Despite the initial response, all animals failed rechallenge and developed new or recurrent tumors within 7 to 10 days. CONCLUSIONS: Coadministration of low-dose IL-2 plus IL-18 induced a potent primary response to murine neuroblastoma likely caused by activation of natural killer cells in the tumor microenvironment. This combined cytokine therapy strategy was unable to induce sustained immunity to rechallenge. However, dendritic cell vaccination combined with IL-2 plus IL-18 cytokine treatment did allow for the establishment of a complete and durable antitumor response. PMID- 12632340 TI - Anti-VEGF antibody in experimental hepatoblastoma: suppression of tumor growth and altered angiogenesis. AB - BACKGROUND: Hepatoblastoma is the most common primary hepatic malignancy of childhood, frequently presenting as advanced disease. Vascular endothelial growth factor (VEGF) is an endothelial mitogen and survival factor critical to growth and angiogenesis in many human cancers. Inhibition of VEGF effectively suppresses tumorigenesis in multiple experimental models. The authors hypothesized that anti VEGF antibody would alter vascular architecture and impede tumor growth in experimental hepatoblastoma. METHODS: The Institutional Animal Care and Use Committee of Columbia University approved all protocols. Xenografts were established in athymic mice by intrarenal injection of cultured human hepatoblastoma cells. Anti-VEGF antibody (100 microg/dose) or vehicle was administered intraperitoneally 2 times per week for 5 weeks. At week 6, 10 control/treated mice were killed and remaining animals maintained without treatment until week 8. Tumor weights were compared by Kruskal-Wallis analysis, and vascular alterations ascertained by fluorescein angiography and specific immunostaining. RESULTS: Anti-VEGF antibody significantly inhibited tumor growth at 6 weeks (1.85 g +/- 0.60 control, 0.05 +/- 0.03 antibody, P <.0003). In comparison with controls, treated xenografts showed decreased vascularity and dilated surviving vessels with prominent vascular smooth muscle elements. CONCLUSIONS: Specific anti-VEGF therapy inhibits neoangiogenesis and significantly suppresses tumor growth in experimental hepatoblastoma. Surviving vasculature displays dilation and increased vascular smooth muscle. Anti-VEGF agents may represent new therapeutic alternatives for children with advanced disease. PMID- 12632342 TI - Excellent outcome in patients with stage I germ cell tumors of the testes: a study of the Children's Cancer Group/Pediatric Oncology Group. AB - BACKGROUND/PURPOSE: The aim of this study was to correlate outcomes in patients with stage I testicular germ cell tumors with compliance to surgical guidelines and to confirm previous single-institution experiences that show excellent disease-free survival rates when treated with orchiectomy alone. METHODS: Sixty three patients were entered into this intergroup study (Children's Cancer Group 8881/Pediatric Oncology Group 9048) between 1990 and 1996. Surgical guidelines for orchiectomy included an inguinal approach, early vascular control of the cord structures, and a hemiscrotectomy if a transscrotal violation occurred. Failure of tumor marker normalization or subsequent elevation suggested advanced disease requiring further surgery and chemotherapy. Survival curves were compared using the log-rank test. RESULTS: The median age was 16 months (range, 1 month to 5.6 years). Progression of disease occurred in 11 patients. The 6-year event-free survival (EFS) was 78.5% + 7%. The overall 6-year survival rate was 100%. Adherence to all guidelines was confirmed in only 43 patients (69%). Overall, there was no statistical significance in event-free survival rates between patients who had a correct surgical approach and all patients who had surgical violations; however, 4 patients with transscrotal violations showed a significantly higher rate of disease recurrence. CONCLUSIONS: Patients with stage I germ cell tumors of the testes have excellent survival rates when treated with surgery alone. Despite numerous surgical guideline violations, event-free survival in this multiinstitutional study was similar to previous single institution reports. All patients with relapse or progression of their disease appear to be cured with further surgical excision and chemotherapy. PMID- 12632341 TI - Malignant retroperitoneal and abdominal germ cell tumors: an intergroup study. AB - BACKGROUND/PURPOSE: This randomized study examined survival (S) and event-free survival (EFS) rates using high-or standard-dose cisplatin-based combination chemotherapy and surgical resection for this subset of germ cell tumors. METHODS: Twenty-six of 317 patients enrolled on the POG 9049/COG 8882 intergroup study for malignant germ cell tumors had abdomen or retroperitoneum as the primary site. Twenty-five of 26 were eligible for inclusion (n = 25). Patients had biopsy or resection at diagnosis and randomization to chemotherapy including etoposide, bleomycin, and either standard-dose (PEB) or high-dose cisplatin (HDPEB). In patients with initial biopsy, delayed resection was planned. RESULTS: Median age was 26 months. There were 14 girls and 11 boys. There were 3 stage I to II, 5 stage III, and 17 stage IV patients. Surgical management included primary resection in 5, resection after chemotherapy in 13, and biopsy or partial resection in 7 patients. Overall 6-year EFS rate was 82.8% +/- 10.9%, and 6-year survival rate was 87.6% +/- 9.3%. By group, 6-year survival rate was 90.0% +/- 11.6% for PEB and 85.7 +/- 14.5% for HDPEB. Deaths include one from sepsis, one from malignant tumor progression, and one from bulky disease caused by benign components despite response of the malignant elements to chemotherapy. CONCLUSIONS: Malignant germ cell tumors arising in the abdomen and retroperitoneum have an excellent prognosis despite advanced stage in most children. Aggressive resection need not be undertaken at diagnosis, but a concerted attempt at complete surgical removal after chemotherapy is important to distinguish viable tumor from necrotic tumor or benign elements that will not benefit from further chemotherapy. PMID- 12632343 TI - Increased microvascular density predicts relapse in Wilms' tumor. AB - BACKGROUND/PURPOSE: Tumor stage and histology are the most important prognostic criteria in Wilms' tumors; however, a subset of patients remains who have favorable histology tumors and unexpectedly relapse. The authors postulated that increased microvascular density (MVD), a hallmark for angiogenesis, could identify patients at risk for relapse. METHODS: A case-control study was used to compare relapse (n = 15) with nonrelapse tumors (n = 35). Tumor MVD was counted in 5 random high-powered fields (hpf) using anti-Factor VIII antibody and expressed as mean vessel count/hpf +/- SEM. MVD and clinical data were evaluated using univariate analysis and student's t test. RESULTS: The relapse group had higher MVD than the nonrelapse group (34.9 +/- 2.9 v 22.4 +/- 2; P <.05). When evaluating the favorable histology (FH) group alone, there was higher MVD in the relapse group (32.4 +/- 2.7 v 19 +/- 1.8; P <.05). MVD was found to be the only predictor of relapse when compared with age, sex, tumor weight, and histology. CONCLUSIONS: These results suggest that increased MVD can identify Wilms' tumor patients at high risk for relapse, especially those patients with favorable histology tumors. A larger study is warranted to determine the potential utility of MVD in stratification of Wilms' tumor patients. PMID- 12632344 TI - Epithelial ovarian tumors in children: a retrospective analysis. AB - BACKGROUND/PURPOSE: Epithelial tumors of the ovary account for about 15% of pediatric ovarian masses. The authors reviewed a 14-year experience with ovarian masses to understand the spectrum of pathology, presentation, and outcome of children with epithelial lesions. METHODS: All ovarian masses resected or biopsied at the authors' institution from 1988 to the present were reviewed retrospectively. Patient age, presenting symptoms, operative procedures, postoperative treatment, and outcome were obtained from the medical record. RESULTS: A total of 240 cases were identified. A total of 123 (51.2%) were nonneoplastic in nature. Of the 117 neoplastic masses, 79 (67.5%) were germ cell tumors and nineteen (16.2%) were epithelial-derived tumors. Patients with epithelial tumors had a mean age of 13.9 +/- 4 years. Ten patients (52.6%) presented with abdominal pain, and 9 (47.4%) presented with an asymptomatic mass. Eleven underwent oophorectomy, 6 underwent cystectomy, and 2 had biopsy alone. Four patients (21%) had ascites, and 1 (5.3%) had a pleural effusion. The histopathologic diagnoses for the epithelial tumors included 9 serous cystadenomas (47%) and 3 mucinous cystadenomas (16%), 3 mucinous cystadenocarcinomas (16%), and 4 serous tumors of borderline malignancy (21%). Two patients (11%) had bilateral disease. Four patients (21%) underwent a subsequent laparotomy for either staging or recurrence, and 2 patients (11%) required chemotherapy. One patient (5.3%) died of ovarian adenocarcinoma. CONCLUSIONS: Epithelial tumors comprise a small but significant proportion of pediatric ovarian masses. The pediatric surgeon must understand the biologic characteristics, operative management, and follow-up treatment of these tumors, and how these differ from germ cell lesions. PMID- 12632346 TI - Pediatric neurofibrosarcoma. AB - BACKGROUND/PURPOSE: Neurofibrosarcoma is rare in children, and the natural history and prognostic factors are not well described. The authors present a 57 year review of their experience. METHODS: The charts of children with neurofibrosarcoma were reviewed retrospectively. Statistical analysis was performed using the Chi2 and unpaired t tests. RESULTS: From 1944 to 2001, 38 patients under the age of 21 were diagnosed with neurofibrosarcoma. Twenty-two were boys. The average age at diagnosis was 13.8 years (range, 3 to 19.9 years). Nineteen patients (50%) had neurofibromatosis. The tumor site was as follows: extremity, 19 patients; trunk, 9 patients; head and neck, 7 patients; and retroperitoneum, 3 patients. The average tumor size was 10 cm. The margins after resection were as follows: grossly positive, 9; microscopically positive, 5; negative, 21; and unknown, 3. Patients with positive margins had a 22% survival rate, whereas those with negative or unknown margins had a 34% survival rate. Thirty-two patients achieved a complete response, 2 a partial response, and 4 progressed while on therapy. Twenty-six patients relapsed after a complete response (11 local, 10 distant, 5 both). Of the 15 patients with a distant relapse, 73% (11) relapsed in the lung. Twelve (32%) patients survived with an average follow-up of 14 years (range, 0.3 to 28 years). Nine patients were treated with chemotherapy, 9 with radiation, and 9 with both chemotherapy and radiation. Outcome was not significantly affected by gender, presence of neurofibromatosis, site, margin, or use of adjuvant therapy. CONCLUSION: Neurofibrosarcoma remains a rare disease in children with insufficient contemporary numbers to assess efficacy of therapy. Prognosis remains poor with a high incidence of relapse, particularly in the lungs, suggesting that more aggressive therapies to control both local and distant relapses are needed. PMID- 12632345 TI - Wilms' tumor growth is suppressed by antiangiogenic pigment epithelium-derived factor in a xenograft model. AB - BACKGROUND/PURPOSE: Pigment epithelium-derived factor (PEDF), a potent endogenous inhibitor of angiogenesis, is highly expressed in the kidney. The authors postulated that systemic administration of PEDF would decrease Wilms' tumor growth in a xenograft model, and increased renal vascularity would result in a mouse null for PEDF. METHODS: Tumors were induced in athymic mice using human anaplastic Wilms' tumor cells. Purified PEDF protein or vehicle was administered for 7 days beginning 2 to 3 weeks after inoculation. Tumors were stained with anti-PEDF and anti-Factor VIII antibodies. Mitoses and microvascular density (MVD) were counted per high-power field (hpf). PEDF-null mice were generated on a SV129/C57Bl6 background. Wild-type and null kidneys were assessed for MVD. RESULTS: Mean tumor weight in the 2-week group was 60% less than controls (P <.05). The MVD and mitotic count in treated tumors were significantly less than controls (P <.05). PEDF stained strongly in normal kidneys but was minimal to absent in Wilms' tumor. PEDF-null kidneys had increased MVD compared with wild type (P <.05). CONCLUSIONS: PEDF is expressed strongly in normal murine kidney, and loss of its angioinhibitory activity may contribute to pathologic angiogenesis in Wilms' tumor. Systemic PEDF suppresses WT growth by targeting both the tumor cells and its associated vasculature. PMID- 12632347 TI - Prognostic factors and surgical treatment guidelines for children with rhabdomyosarcoma of the perineum or anus: a report of Intergroup Rhabdomyosarcoma Studies I through IV, 1972 through 1997. AB - BACKGROUND/PURPOSE: Rhabdomyosarcoma (RMS) of the perineum or anus is a rare sarcoma of childhood with a poor prognosis. This study reviews the Intergroup Rhabdomyosarcoma Study Group (IRSG) studies I through IV to identify determinants of patient outcome and to refine surgical treatment guidelines. METHODS: From 1972 through 1997, 71 eligible patients were treated and studied. The median patient age was 6 years. The majority (64%) were at an advanced stage (clinical group III and IV) at initial presentation and 50% had positive regional lymph node (LN) involvement. RESULTS: The 5-year failure-free survival rate (FFS) for all patients was 45% and the overall survival rate (OS) was 49%. Characteristics that were associated with significantly improved survival rate were primary tumor size less than 5 cm, lower (less advanced) clinical group and stage, negative regional lymph node status, and age less than 10 years. When the extent of disease was controlled for in multivariate analysis, only age less than 10 predicted an improved outcome. The 5-year overall survival rate for patients less than 10 years of age was 71% versus 20% in older patients (P <.001). Histology (alveolar versus embryonal) and primary site (perineal versus anal) did not significantly affect outcome. CONCLUSIONS: Because of the high incidence of regional LN involvement in these patients, a strategy of routine surgical evaluation of ilioinguinal lymph nodes in all patients with perineal or anal RMS is recommended. PMID- 12632348 TI - Serum amylase and lipase alone are not cost-effective screening methods for pediatric pancreatic trauma. AB - BACKGROUND: Injury to the pancreas is rare in pediatric trauma. Identification of pancreatic injury relies on clinical, radiographic, and laboratory data. Serum screening for pancreatic injury frequently is used but has not proven to correlate well with pancreatic injury. This study investigated utility and cost effectiveness of serum assessment of amylase and lipase. METHODS: A retrospective study of 1,821 pediatric trauma patients over 64 months was conducted. A total of 293 (16%) of these patients suffered trauma to the torso 195 (11%) of whom had confirmed intraabdominal injury. Eight pancreatic injuries (4% of abdominal injuries) were identified; 5 underwent surgery for pancreatic ductal injury. One patient not operated on had a pseudocyst that required late drainage. RESULTS: Serum amylase or lipase levels (AMY/LIP) were measured in 507 (28%) patients. A total of 116 (23%) had elevated AMY/LIP levels. Six of 8 with proven pancreatic injury underwent AMY/LIP testing; 5 had elevated values. Forty-eight percent of patients with elevated AMY/LIP levels had no evidence of intraabdominal injury. Seventy-four of 116 (64%) with elevated AMY/LIP levels underwent abdominal and pelvic computed tomography (CT) scanning, yet 38 (51%) of these had completely normal scans. Many patients with elevated AMY/LIP levels (cost, $6 per test) underwent screening CT scans (cost, $592 per test) based on AMY/LIP alone. No patient with elevated AMY/LIP levels but without clinical suspicion was proven to have pancreatic injury. Cost data are presented. CONCLUSIONS: Serum amylase and lipase determinations may support clinical suspicion in the diagnosis of pediatric pancreatic trauma but are not reliable or cost effective as screening tools. Costs incurred from routine serum amylase and lipase or from imaging tests subsequent to elevated serum values may be significant and unjustified. PMID- 12632349 TI - A multidisciplinary approach to the development of a cervical spine clearance protocol: process, rationale, and initial results. AB - BACKGROUND/PURPOSE: Assessment of potential spine injuries is inconsistent and controversial. Subsequent morbidity includes prolonged immobilization and missed injuries. To address these issues, a multidisciplinary team was organized to design a cervical spine management/clearance pathway. The process, algorithm, and initial results are described. METHODS: Team members consisted of pediatric surgeons, orthopedic surgeons, neurosurgeons, emergency room physicians, and trauma nurse practitioners. Nationwide standards, guidelines, and experiences across disciplines were reviewed, and a consensus pathway evolved for cervical spine clearance in children 8 years and younger. A short-term retrospective review (5 months) was performed to assess initial performance. Time required for clearance, number and type of imaging studies, and number of missed injuries were compared between a group of patients before (n = 71) and after (n = 56) the implementation of the pathway. RESULTS: Strict guidelines for cervical spine immobilization and clearance criteria were defined. After implementation of this pathway, time required for cervical clearance in nonintubated children decreased (before, 12.3 +/- 1.5 v after, 7.5 +/- 0.9 hours; P =.014). A clear trend toward earlier clearance in intubated patients existed (before [n = 6], 40.0 +/- 16.8 v after [n = 6], 19.4 +/- 8.1 hours; P =.10); there need to be larger numbers to determine statistical significance. The 2 study groups were similar in age; mechanism of injury; Glasgow coma scale score; and number of plain x-rays, computed tomography scans, and magnetic resonance imaging studies obtained. Neither group had missed injuries. CONCLUSIONS: standards for cervical spine immobilization, assessment, and clearance. Implementation of such guidelines decreased time for cervical spine clearance, and ongoing analysis of sensitivity is encouraging. PMID- 12632350 TI - Significance of 'blush' on computed tomography scan in children with liver injury. AB - BACKGROUND/PURPOSE: The aim of this study was to determine if the presence of "blush" (an indication of active bleeding) on abdominal CT in children with blunt liver injury adversely affected their clinical outcome as has been reported in adults. METHODS: The authors reviewed the records of 105 children ages 1 to 16 years with blunt liver injury seen on admission IV contrast CT seen over a 6-year period. Demographic characteristics measured were age, mechanism of injury, and injury severity score (ISS). Clinical outcomes included ICU stay, hospital length of stay (LOS), transfusion requirement (milliliters per kilogram), operations performed, and mortality rate. CT scans were evaluated retrospectively by a radiologist blinded to prior reports, for a "blush" and grade of liver injury. No patient underwent arterial embolization. The authors eliminated children with grade I-II injuries (30 patients), because only one had a blush, and analyzed the 75 patients with severe liver injuries (grades III-V). Those patients without a blush (n = 53) seen on CT were the control group, whereas patients with a blush (n = 22) were the study group. Data were analyzed using the Fisher's Exact and Mann-Whitney U test. The level of significance was set at.05. RESULTS: Patients with a blush had a significantly larger transfusion requirement (17.3 +/- 30.5 mL/kg v 5.0 +/- 10.9 mL/kg; P =.02) and mortality rate (23% v 4%; P =.02), but the ISS also was significantly greater (25.8 +/- 14.5 v 17.5 +/- 12.2; P =.019). All other data were similar between the 2 groups. CONCLUSIONS: Children with a blush seen on abdominal CT after blunt liver injury have higher transfusion requirements and greater risk of mortality than those without blush. Mortality is primarily related to the severity of their other injuries. PMID- 12632351 TI - The diagnosis of acute appendicitis in a pediatric population: to CT or not to CT. AB - PURPOSE: The aim of this study was to determine if focused appendiceal computed tomography with colon contrast (FACT-CC) increases the accuracy of the preoperative diagnosis of acute appendicitis in children. METHODS: A 5-year retrospective review was conducted of a university hospital database of 283 patients (age 0.8 to 19.3 years; mean, 11.3 years) treated with appendectomy for presumed acute appendicitis. RESULTS: Of the 283 patients in whom appendectomies were performed, 268 were confirmed by pathologic analysis of the specimen to have acute appendicitis for a diagnostic accuracy in our institution of 94.7%. Ninety six patients (34%) underwent FACT-CC scans as part of their preoperative evaluation. The sensitivity of the computed tomography (CT) scan was 94.6%, and the positive predictive value was 95.6%. In girls older than 10 years, CT imaging was not significantly more accurate in predicting appendicitis than examination alone (93.9% v. 87.5%; P =.46). CONCLUSIONS: Preoperative FACT-CC did not increase the accuracy in diagnosing appendicitis when compared with patients diagnosed by history, physical examination and laboratory studies. If there was a strong suspicion of appendicitis, a negative CT scan did not exclude the diagnosis of appendicitis. However, focused appendiceal CT scan is a sensitive test with a high positive predictive value and may be useful in a patient with an atypical history or examination. PMID- 12632353 TI - Cardiopulmonary effects of closed repair of pectus excavatum. AB - BACKGROUND/PURPOSE: Increasing numbers of patients with pectus excavatum defects are presenting for operative repair. Studies that follow-up with patients after open repair have found a decrease in pulmonary function with some improvement in cardiac output and exercise tolerance; however, these effects have not been examined systematically after closed or Nuss repair of pectus excavatum. This study examined the early postoperative effects of closed repair of pectus on pulmonary function, exercise tolerance, and cardiac function. METHODS: Patients were followed up prospectively after initial evaluation for operation. All patients underwent preoperative computed tomography (CT) scan, and pre- and postoperative (3 months) pulmonary function studies, exercise tolerance, and echocardiographic evaluation of cardiac function. RESULTS: Eleven patients underwent evaluation. Preoperative CT index was 4.1 +/- 0.9. Patients reported an improvement in subjective postoperative exercise tolerance (4.1 +/- 0.7; maximal, + 5). Pulmonary function studies (FVC and vital capacity) were significantly reduced at 3 months postsurgery: change in FVC, -0.67 +/- 0.92 L and VC, -0.5 +/- 0.72 L. Similarly, VO2 max was reduced: preoperative, 35.6 +/- 1.5 versus postoperative, 29.1 +/- 11.9 L/kg/min. Cardiac function was significantly improved postoperation (stroke volume preoperative, 61.6 +/- 25 versus 77.5 +/- 23 mL postoperative). All comparisons had a P value less than.05 by Student's paired t test. CONCLUSIONS: These results show that closed repair of pectus excavatum is associated with a subjective improvement in exercise tolerance, which is paralleled by an increase in cardiac function and a decline in pulmonary function. These findings support the use of closed repair of pectus excavatum in patients who complain of subjective shortness of breath; further study is required to delineate the long-term cardiopulmonary implications after closed repair. PMID- 12632352 TI - Appendicitis 2000: variability in practice, outcomes, and resource utilization at thirty pediatric hospitals. AB - BACKGROUND/PURPOSE: To improve clinical results and resource utilization in the care of appendicitis in children, the authors examined the current practice and outcomes of 30 pediatric hospitals. METHODS: The Pediatric Health Information System (PHIS) database consists of comparative data from 30 free-standing Children's hospitals. The study population of 3,393 children was derived from the database by selecting the "Diagnosis Related Group Code" for appendicitis (APRDRGv12 164), ages 0 to 17 years, using discharges between October 1, 1999 and September 30, 2000. Data are expressed as the range and median for individual hospital outcomes. RESULTS: The nonpositive appendectomy rate ranged from 0 to 17% at the 30 hospitals (median, 2.6%). Ruptured appendicitis varied from 20% to 76% (median, 36.5%). The median length of stay (LOS) for nonruptured appendicitis was 2 days (range, 1.4 to 3.1 days), ruptured appendicitis varied from 4.4 to 11 days (median, 6 days). The median readmission rate within 14 days was 4.3% (0 to 10%). Laparoscopic appendectomy varied from 0 to 95% in the 30 hospitals (mean, 31%) The LOS did not vary significantly in laparoscopic versus open for nonruptured (2.3 v 2.0 days) or ruptured appendicitis (5.5 v 6.2 days). Days on antibiotics for ruptured appendicitis ranged from 4.6 to 7.9 days (median, 5.9 days) Children receiving any study varied from 18% to 89% (median, 69%). Ultrasound scan and computed tomography (CT) were comparable in both nonruptured (13% ultrasound scan v 14%) and ruptured appendicitis (14% ultrasound scan v 21% CT). CONCLUSIONS: Significant variability in practice patterns and resource utilization exists in the management of acute appendicitis in pediatric hospitals. Clinical outcomes could be improved by collaborative initiatives to adopt evidence-based best practices. PMID- 12632354 TI - Monitoring intraoperative effectiveness of caudal analgesia through skin temperature variation. AB - BACKGROUND/PURPOSE: A reliable noninvasive intraoperative marker of caudal analgesia effectiveness remains elusive. Caudal analgesia causes sympathetic inhibition resulting in vasodilatation, increased blood flow, and a resultant increase in temperature in the affected dermatomes. The authors hypothesize that this change in temperature between the affected and unaffected dermatomes is measurable and may represent a noninvasive method of monitoring effectiveness of caudal analgesia. METHODS: Children undergoing lower abdominal surgery participated in the caudal or noncaudal (control) analgesia arm of the study. After induction of general anesthesia, 0.25% bupivicaine (1 mL/kg) was infiltrated for a field block in control patients or a caudal block in the experimental group. Temperature was measured at the C4 and L2 dermatomes starting after induction and 5 minutes before the caudal or field block and every 5 minutes after. T(o) is defined as the difference between the C4 and L2. Delta T (DeltaT) is the temperature variation between T(o). A change in the DeltaT is defined by an increase in the L2 temperature. RESULTS: Forty-six families enrolled (36 experimental, 10 control). The DeltaT for controls was 0.2+/-0.09 degrees C (SEM). Each child in the experimental group had 2 temperature measurements before the caudal with an average DeltaT of 0.3+/-0.07 degrees C (SEM), thus, were internal controls. A marked increase in DeltaT at 5 minutes 0.5+/-0.06 degrees C (SEM) and at 10 minutes 0.6+/-0.07 degrees C (SEM; P <.05) was noted in the experimental group. CONCLUSION: A significant transient change in temperature is observed after caudal analgesia and maybe a noninvasive marker of effectiveness. Further study may clarify its clinical utility. PMID- 12632355 TI - Development of a quality assessment scale for retrospective clinical studies in pediatric surgery. AB - BACKGROUND/PURPOSE: More than 97% of the clinical research in pediatric surgery consists of retrospective data. Although these studies have significant limitations, they fundamentally shape clinical practice within the field. In this report, the authors describe the development and potential applications of a standardized quality assessment scale designed for retrospective studies in pediatric surgery. METHODS: The authors developed a comprehensive quality assessment instrument incorporating 30 items within 3 subscales. These subscales were designed to assess 3 integral components of study design: clinical relevance, reporting methodology, and the strength of conclusions. Global quality ratings (poor, fair, or good) were derived by combining scores from each subscale. To examine inter-rater reliability, 10 retrospective studies from the pediatric surgery literature were assessed with the instrument by 6 independent reviewers. RESULTS: Inter-rater reliability was excellent as assessed by the level of agreement for all items within the instrument (84.6% concordance, n = 1,573 total item-choices) and for individual subscales (range, 73.3% to 85.8%, n = 60 to 1,258). The extent of agreement among reviewers was 82% for classifying reports into qualitative categories by global quality scores (n = 60). The ranking of papers by individual reviewers was highly predictive of overall ranking by mean quality scores (n = 60 rank pairs, r = 0.83; P <.01). CONCLUSIONS: The authors have developed a standardized and reliable quality assessment scale for the analysis of retrospective data in pediatric surgery. Potential applications include: (1) providing the practicing surgeon with a knowledge base to critically evaluate published retrospective data; (2) providing a standardized methodology for the systematic review of existing retrospective data; and (3) developing standardized reporting guidelines for use in peer reviewed journals. PMID- 12632356 TI - The market for pediatric surgeons: an updated survey of recent graduates. AB - PURPOSE: The aim of this study was to identify the demand for pediatric surgeons as perceived by recent graduates of North American pediatric surgery training programs. METHODS: A survey was mailed to every pediatric surgeon who completed a certified training program in North America between 1998 and 2000; 81% of 83 responded. Data from the previous survey of 1992 through 1997 graduates provided longitudinal comparisons. The data were analyzed using univariate and bivariate statistical analysis. RESULTS: The number of graduates was 28, 31, and 24 for 1998, 1999, and 2000, respectively, down from 35 graduates in 1997. All found positions in pediatric surgery, and first-year incomes continue to rise. Fewer graduates (54%) desired an academic position coupled with research compared with previous survey respondents (64%), and 69% were able to find such a position. A practice at a university children's hospital remained the most desired by over 80%, but only 38% obtained positions in this type of practice. However, 55% were able to find a position in a children's hospital (university or community), which had declined gradually to a low of 32% in 1997. The 1998 to 2000 graduates perceive a much stronger demand for pediatric surgeons compared with previous survey respondents (46% v 14%), and greater than 50% felt that too few pediatric surgeons were being trained. CONCLUSIONS: The market demand for pediatric surgeons remains strong as measured by employment and income. The perception of the market by the 1998 through 2000 graduates is positive, which contrasts with the 1992 through 1997 graduates. However, there continues to be significant market changes including a proliferation of positions at community Children's hospitals. PMID- 12632357 TI - High-dose steroids, ursodeoxycholic acid, and chronic intravenous antibiotics improve bile flow after Kasai procedure in infants with biliary atresia. AB - BACKGROUND/PURPOSE: Early reports suggest that the use of steroids after Kasai portoenterostomy may improve bile flow and outcome in infants with biliary atresia. METHODS: Of 28 infants with biliary atresia, half received adjuvant high dose steroids, and half received standard therapy. Infants in the steroid group (n = 14) received intravenous solumedrol (taper of 10, 8, 6, 5, 4, 3, 2 mg/kg/d), followed by 8 to 12 weeks of prednisone (2 mg/kg/d). The steroid protocol also included ursodeoxycholic acid indefinitely and intravenous antibiotics for 8 to 12 weeks followed by oral antibiotic prophylaxis. Infants in the standard therapy group (n = 14) received no steroids, occasional ursodeoxycholic acid, and perioperative intravenous antibiotics followed by oral antibiotic prophylaxis. The infants were not assigned randomly, but rather received standard therapy or adjuvant steroid therapy according to individual surgeon preference. RESULTS: Eleven of 14 (79%) in the steroid group and 3 of 14 (21%) in the standard therapy group had a conjugated bilirubin level less than 1.0 within 3 to 4 months of surgery (P <.001). Fewer patients in the steroid group (21% v 85%) required liver transplantation or died during the first year of life (P <.001). Infants in the steroid group did better despite the fact that this group included 5 infants with biliary atresia-polysplenia-heterotaxia syndrome, a subgroup that might have been expected to have a poor prognosis. Neither bile duct size nor liver histology was a reliable predictor of success or failure in either group. CONCLUSIONS: Adjuvant therapy using high-dose steroids, ursodeoxycholic acid, and intravenous antibiotics may accelerate the clearance of jaundice and decrease the need for early liver transplantation after Kasai portoenterostomy. PMID- 12632359 TI - MUC-2 mucin production in Hirschsprung's disease: possible association with enterocolitis development. AB - BACKGROUND/PURPOSE: The etiology of Hirschsprung's-associated enterocolitis (HAEC) is unknown. Previous investigations have suggested that abnormal production of mucins may have an etiologic role. Recently, a series of mucin genes have been identified. MUC-2 is the predominant mucin expressed in humans. The authors have shown previously in vitro that use of MUC-2 can prevent bacterial translocation. Based on this, it was hypothesized that those patients with Hirschsprung's disease (HD) would have an abnormal production of MUC-2 compared with normal patients. METHODS: Fresh stool specimens were collected from children with a diagnosis of HD (with or without HAEC) and from age-matched control patients. Protein was extracted, and MUC-2 was detected with Western blot analysis. MUC-2 protein expression was quantified by densitometry measurements. Results are expressed as mean density +/- SD. Statistical comparison was done with unpaired t tests, with P less than.05 being considered significant. RESULTS: MUC-2 expression was detected in all control patients (mean density, 121 +/- 47). MUC-2 level was lowest in one child with a viral-induced diarrhea (density = 71). In those patients with HD, levels of MUC-2 protein expression were significantly lower (P <.05) than controls (12 +/- 15 for all HD patients). Levels of MUC-2 were lowest (nondetectable) in 2 HD patients who had clinical evidence of HAEC. CONCLUSIONS: MUC-2 production is markedly depressed in patients with Hirschsprung's disease and is absent with enterocolitis. This decline in protein expression may result in a decrease in epithelial barrier function and be a predisposing factor in the development of HAEC. PMID- 12632358 TI - Diagnosing Hirschsprung's disease: increasing the odds of a positive rectal biopsy result. AB - BACKGROUND/PURPOSE: Rectal biopsies are performed as a definitive means of diagnosing Hirschsprung's Disease (HD) in children presenting with constipation. The authors hypothesized that key features in the history, physical examination, and radiographic evaluation would allow us to avoid unnecessary rectal biopsies. METHODS: A retrospective analysis was conducted on patients undergoing rectal biopsy between 1995 and 2001. Patients with HD were identified (n = 50), and a concurrent cohort of patients with idiopathic constipation (IC; n = 50) was selected. Pertinent features in patients with HD versus those with IC were cross tabulated using Pearson Chi2 testing (significance was P <.05). RESULTS: Sixty percent of patients with HD and 15% of patients with IC experienced onset of symptoms in the first week of life. HD patients more frequently experienced delayed passage of meconium (P <.05), abdominal distension (P <.05), vomiting (P <.05), and transition zone on contrast enema (P <.05). All patients with HD had one or more of these significant features. In contrast, only 64% of patients with IC had one or more of these features. The classic triad of symptoms (ie, delayed passage of meconium, vomiting, and abdominal distension) was present in 18%, and one or more of these symptoms was present in 98% of HD patients. In contrast, only 60% of patients with IC had a history of delayed passage of meconium, vomiting, or abdominal distension. CONCLUSIONS: A history of delayed passage of meconium, abdominal distension, vomiting or the results of a contrast enema identified all patients with HD and excluded HD in approximately 36% of patients with idiopathic constipation. The authors have shown that key features in a patient's history, physical examination, and radiologic evaluation can differentiate between HD and IC. In a child presenting with constipation and none of the above features, it is not necessary to perform a rectal biopsy to exclude HD. PMID- 12632360 TI - Improved quality of life by combined transplantation in Hirschsprung's disease with a very long aganglionic segment. AB - BACKGROUND/PURPOSE: The treatment of children with Hirschsprung's disease beginning in the proximal jejunum remains a challenge for the pediatric surgeon. These patients need a definitive parenteral nutrition, which could lead to a liver impairment. The goal of this work is to assess the quality of life after combined liver, intestine, and right colon transplantation. METHODS: This is a retrospective study of 3 patients. Data regarding symptomatology, radiographic and operating findings, postoperating recovery, and quality of life were analyzed and compared with the quality of life before the transplantation. RESULTS: The suspicion of a very long intestinal aganglionosis should be derived from the intestinal biopsies. Three combined liver, intestine, and right colon transplantation operations have been performed. The immunosuppression included steroids, tacrolimus, and azathioprine. An abdominal pull-trough (Duhamel procedure 2, Swenson procedure 1) was performed from 6 to 24 months after the transplantation. The follow-up after the transplantation ranges from 2 to 6 years. These 3 patients are completely off total parenteral nutrition with bowel movements 2 to 3 times a day. Two patients are continent day and night, and one is continent during the day only. CONCLUSIONS: Intestinal transplantation is feasible with good results even when a liver impairment needs a combined intestine and liver transplantation. The right colon transplantation, in our experience, does not impair the results. The quality of life after the transplantation is better than before. PMID- 12632361 TI - Serial transverse enteroplasty (STEP): a novel bowel lengthening procedure. AB - BACKGROUND/PURPOSE: Bowel lengthening may be beneficial for children with short bowel syndrome. However, current techniques require at least one intestinal anastomosis and place the mesenteric blood supply at risk. This study seeks to establish the technical principles of a new, simple, and potentially safer bowel lengthening procedure. METHODS: Young pigs (n = 6) underwent interposition of a reversed intestinal segment to produce proximal small bowel dilation. Five weeks later the reversed segment was resected. Lengthening of the dilated bowel then was performed by serial transverse applications of a GIA stapler, from opposite directions, to create a zig zag channel. A distal segment of equal length served as an in situ morphometric control. Contrast radiologic studies were performed 6 weeks later, and the animals were killed. Statistical comparisons were made by paired t test with P less than.05 considered significant. RESULTS: After bowel lengthening, all animals gained weight (66.7 +/- 3.0 [SD] kg v 42.5 +/- 3.5 kg; P <.001) and showed no clinical or radiologic evidence of intestinal obstruction. Intraoperatively, immediately after serial transverse enteroplasty, the intestine was substantially elongated (82.8 +/- 6.7 cm v 49.2 +/- 2 cm; P <.01). Six weeks after surgery, the lengthened intestinal segment became practically straight and, compared with the in situ control, remained significantly longer (80.7 +/- 13.1 cm v 57.2 +/- 10.4 cm; P <.01). There was no difference in diameter between these segments (4.3 +/- 0.7 cm v 3.8 +/- 0.4 cm; P value, not significant). CONCLUSIONS: Serial transverse enteroplasty (STEP) significantly increases intestinal length without any evidence of obstruction. This procedure may be a safe and facile alternative for intestinal lengthening in children with short bowel syndrome. PMID- 12632362 TI - Laparoscopic Roux-en-Y gastric bypass in morbidly obese adolescents. AB - BACKGROUND/PURPOSE: Obesity has contributed significantly to morbidity and premature deaths in the adolescent population. Because many patients do not respond to dietary modification, exercise regimens, or pharmacologic treatment, weight reduction surgery has become a viable alternative, although the morbidity of conventional gastric bypass has tempered enthusiasm for this approach. Experience with the laparoscopic approach has not been reported previously. The authors examined the outcome of adolescents undergoing laparoscopic Roux-En-Y gastric bypass (lap RYGB). METHODS: Medical records of patients less than 20 years of age (n = 4; 3 girls, 1 boy) who had undergone lap RYGB for morbid obesity were reviewed. All patients met National Institute of Health criteria for bariatric surgery. Outcome variables examined included weight; body mass index (BMI); hospital length of stay (LOS); comorbid conditions; and tolerance of a regular diet. Mean time to follow-up was 17 months. RESULTS: All procedures were completed laparoscopically. There were no complications. The average LOS was 2 days. Patients with greater than 20-month follow-up lost an average of 87% of their excess body weight and had nearly complete resolution of comorbidities (including hypertriglyceridemia, hypercholesterolemia, asthma, and gastroesophageal reflux disease). CONCLUSION: Laparoscopic gastric bypass is a safe alternative in morbidly obese adolescents who have not responded to medical therapy. PMID- 12632363 TI - Heparin-binding EGF-like growth factor downregulates expression of adhesion molecules and infiltration of inflammatory cells after intestinal ischemia/reperfusion injury. AB - BACKGROUND/PURPOSE: This study examined whether heparin-binding epidermal growth factor (EGF) like growth factor (HB-EGF), a proven intestinal cytoprotective molecule, exerts its protective effects through modulation of adhesion molecule expression and inflammatory cell infiltration, important pathogenic mediators of ischemia/reperfusion (I/R) injury. METHODS: Total midgut I/R injury in rats was achieved by occlusion of the superior mesenteric artery for 90 minutes followed by reperfusion. Rats were treated intraluminally with 600 microg/kg HB-EGF or with PBS 45 minutes after the onset of ischemia. Four- or 24-hours post-I/R, ileum was harvested and processed for immunhistochemical detection of P-/E selectins, intercellular adhesion molecule-1 (ICAM-1)/vascular cell adhesion molecule-1 (VCAM-1), and polymorphonuclear cells (PMN)/macrophages (MPhi). RESULTS: P-/E-selectins were significantly induced in vascular endothelia 4 hours after I/R injury compared with normal intestine. HB-EGF treatment significantly down-regulated the expression of P-/E-selectins. I/R-injured intestine displayed overexpression of ICAM-1 and VCAM-1, which were significantly down-regulated by HB-EGF treatment. Lastly, I/R injury caused significant infiltration of PMN and MPhi into wounded tissue 24 hours after I/R compared with normal intestine. HB EGF treatment significantly decreased PMN and MPhi infiltration into the injured tissue. CONCLUSIONS: HB-EGF intestinal cytoprotection is mediated, in part, by down-regulation of expression of adhesion molecules and infiltration of PMN and MPhi after intestinal I/R injury. PMID- 12632364 TI - Localization of postresection EGF receptor expression using laser capture microdissection. AB - BACKGROUND/PURPOSE: Epidermal growth factor (EGF) and its receptor (EGFR) are key components in the genesis of adaptation after small bowel resection (SBR). Within intestinal homogenates, EGFR expression is increased after SBR; however, the exact cells responsible for altered EGFR expression are unknown. In this study, laser capture microdissection (LCM) microscopy was used to elucidate the specific cellular compartment(s) responsible for postresection changes in EGFR expression. METHODS: Male ICR mice underwent a 50% proximal SBR or sham operation. After 3 days, frozen sections were taken from the remnant ileum. Individual cells from villi, crypt, muscularis, and mesenchymal compartments were isolated by LCM. EGFR mRNA expression for each cell compartment was quantified using real-time polymerase chain reaction (PCR). RESULTS: EGFR expression was increased after SBR within the crypt (2-fold) and muscularis compartments (3-fold). There were no changes detected after SBR in the villus tips or mesenchymal compartments. CONCLUSIONS: Increased expression of EGFR in crypts directly correlates with the zone of cell proliferation and supports the hypothesis that EGFR signaling is crucial for the mitogenic stimulus for adaptation. The finding of increased EGFR expression in the muscular compartment is novel and may implicate a role for EGFR as a mediator of the muscular hyperplasia seen after massive SBR. PMID- 12632366 TI - Correction of hindbrain herniation and anatomy of the vermis after in utero repair of myelomeningocele in sheep. AB - BACKGROUND/PURPOSE: In utero repair of myelomeningocele (MMC) in humans spares distal neurologic function, reverses the hindbrain herniation component of the Arnold-Chiari II malformation (ACM), and reduces the rate of postnatal shunt placement. The authors hypothesized that extravasation of cerebrospinal fluid (CSF) from the lumbar spinal cord results in herniation. This hypothesis was tested by assessing the impact of a spinal cord myelotomy on hindbrain anatomy in fetal sheep. METHODS: A MMC lesion was created surgically in 34 fetal sheep at 75 days' gestation by excision of the L1-L5 lamina, the exposed dura, and surrounding tissues. A lumbar level myelotomy was performed in 28 of the 34 fetuses to open the central canal of the spinal cord to enhance egress of CSF through the MMC defect and potentially induce hindbrain herniation. At 102 days' gestation, a repair of the MMC lesion was performed in 14 fetuses with a myelotomy. Fetuses underwent autopsy at 102, 114, 120, or 140 days' gestation. Control animals underwent 2 unrelated fetal surgical procedures at approximately 70 and 110 days' gestation. The incidence of hindbrain herniation, ventricular size, biparietal diameter, brain weight, and brain anatomy were compared between the different animal groups. RESULTS: After MMC creation, significant cerebellar tonsillar herniation was observed in 85% of fetuses that underwent creation of a myelotomy; none of the lambs without a myelotomy (n = 6) had hindbrain herniation. At autopsy, cerebellar tonsillar herniation was present at the time of MMC repair (102 days' gestation), 2 weeks after MMC repair, but was reversed 3 weeks post-MMC repair. At birth, tonsillar herniation was absent, and hindbrain anatomy was restored in 88% of the fetuses with a myelotomy that underwent fetal MMC repair. No significant differences in brain weight and ventricular size was observed between animals with and without MMC repair. CONCLUSIONS: Adding a myelotomy to the sheep model of MMC leads to hindbrain herniation that is similar to that observed in the human ACM. These experiments support the hypothesis that leakage of CSF through the exposed central canal alters the normal CSF hydrodynamics, resulting in cerebellar tonsillar herniation. Fetal MMC repair reverses hindbrain herniation and restores gross anatomy of the vermis. PMID- 12632365 TI - Spectrum of intrapartum management strategies for giant fetal cervical teratoma. AB - BACKGROUND/PURPOSE: The management of the fetus with a large neck mass that obstructs the airway remains a clinical challenge. The authors review their experience with giant fetal cervical teratoma and discuss options for management. METHODS: A retrospective review of all patients referred since 1994 for prenatal management of a fetal neck mass was performed. Variables examined included gestational age at diagnosis and delivery, size and location of the neck mass, presence of fetal hydrops, associated anomalies, management methods, operating time, and outcome. RESULTS: Seven patients were identified with a prenatal diagnosis of giant cervical teratoma. Four patients had fetal hydrops; of these, 2 died in utero of hydrops, and a third fetus underwent elective termination. The remaining hydropic and previable fetus underwent fetal surgery for resection of the mass. The 3 nonhydropic patients underwent ex utero intrapartum treatment (EXIT) procedures for airway control. Endotracheal intubation was possible in one patient, and one received a tracheostomy. In the third fetus, neither intubation nor tracheostomy were possible, and resection of the neck mass was performed on placental support. There were no deaths in the surgical group. CONCLUSIONS: The management of fetal giant cervical teratoma includes a spectrum of options. For the rare previable fetus with hydrops, fetal resection may be indicated. In patients with airway obstruction, EXIT procedure provides the luxury of time to obtain airway control either by intubation, tracheostomy, or, if necessary, tumor resection on placental support. PMID- 12632367 TI - Improved outcomes in the treatment of gastroschisis using a preformed silo and delayed repair approach. AB - BACKGROUND/PURPOSE: The aim of this study was to critically evaluate the clinical outcomes of two different surgical treatment approaches for infants born with gastroschisis. METHODS: The medical records of 65 infants with gastroschisis treated at one institution from 1991 to 2000 were available. Infants in group I (prior to December 1998) underwent attempted early repair of the gastroschisis defect on their first day of life. Infants in group II had delayed repair after the initial placement of a preformed silo. RESULTS: Group I had 39 patients; group II had 26 patients. The two groups were equal with respect to maternal age, gestational age, and birth weight. Complete reduction and fascial closure were accomplished for 32 patients (82%) in group I and 25 patients (96%) in group II (P <.02). Median time on the ventilator was significantly less for group II (P <.0001). Infants in group II had shorter times until first postoperative feeding (P <.01) and full feedings (P <.006). Group II had fewer complications than group I (23% v 56%; P <.01). There appeared to be less necrotizing enterocolitis in group II. The average length of hospital stay was 14 days less for group II. CONCLUSIONS: The use of a preformed silo initially followed by delayed fascial closure in infants with gastroschisis is associated with improved fascial closure rates, fewer ventilator days, more rapid return of bowel function, and fewer complications compared with attempts at initial early repair. PMID- 12632369 TI - Notochord anomalies in the adriamycin rat model: A morphologic and molecular basis for the VACTERL association. AB - BACKGROUND/PURPOSE: The Adriamycin rat model (ARM) is a reliable model of the VACTERL association. The notochord is structurally abnormal in the region of the foregut, midgut, and hindgut in the ARM. The authors hypothesised that notochord anomalies allow ectopic expression of molecular signals in the developing embryo and thus lead to VACTERL malformations. This study was designed to investigate this hypothesis. METHODS: Adriamycin (1.75 mg/kg) was administered intraperitoneally to pregnant rats on days 7, 8, and 9 of gestation. Control animals were given saline. Embryos were recovered on gestational days 10.5 to 14 at (1/2)-day intervals and at full term. The first group of embryos were embedded in resin, and sagittal sections stained with Toluidine blue were studied for morphologic abnormalities. The second group of embryos were examined using in situ hybridization for the expression of Sonic Hedgehog (Shh), a patterning gene implicated in the etiology of the VACTERL association. RESULTS: Twenty-seven of the 28 (96.4%) full-term embryos showed VACTERL anomalies. Forty-five of the 50 (90%) experimental embryos (gestational days 10.5 to 14) showed notochord abnormalities. Abnormal ventral branches from the notochord toward the gut were a commonly observed abnormality. These were seen to impinge on the developing foregut, midgut, dorsal aorta, and kidney. In situ hybridization for Shh showed that these branches from the notochord expressed Shh in 66.6% of experimental embryos. This abnormal Shh expression was not seen in the control embryos. CONCLUSIONS: Adriamycin diffusely induces altered notochord morphology in the rat embryo. The abnormal notochord morphology may allow ectopic expression of Sonic Hedgehog, and, thus, contribute to the malformations found in the VACTERL association. PMID- 12632368 TI - A role for sonic hedgehog signaling in the pathogenesis of human tracheoesophageal fistula. AB - BACKGROUND/PURPOSE: Many theories of the pathogenesis of esophageal atresia with tracheoesophageal fistula (EA/TEF) have been proposed, but no specific mechanism has been demonstrated. The authors previously reported data suggesting a respiratory origin of the fistula tract in the rat model and in humans. Sonic hedgehog (Shh) "knockout" mice have the VACTERL association, and thus it was hypothesized that defects in Shh signaling may exist in the human neonatal EA/TEF fistula tract. METHODS: With IRB approval, human proximal esophageal pouch and distal fistula samples were removed at the time of standard repair of EA/TEF in accordance with what the surgeons deemed appropriate in preparation for anastomosis. Tissues were processed for HE, reverse-transcriptase polymerase chain reaction (RT-PCR), and immunohistochemistry. Normal embryonic lung cDNA was used as a positive control for the RT-PCR reactions. RESULTS: As expected, Shh was present by immunohistochemistry in the proximal esophageal pouch, but was specifically absent in the distal fistula tract. Gli-1, -2, and -3 (all intracellular mediators of Shh signaling) were present in the proximal pouch and distal esophagus by RT-PCR. CONCLUSIONS: The absence of Shh signaling in the developing fistula tract of the human neonate was surprising given that Shh normally is present in esophagus and other gut components. These results support the conclusion that the fistula tract is not an esophaguslike structure, despite both its histologic appearance and its use as an esophageal replacement. Also, like in Shh-null mutant mice, aberrant Shh signaling may play a critical role in the pathogenesis of EA/TEF in humans. PMID- 12632371 TI - Cardiopulmonary performances in young children and adolescents born with large abdominal wall defects. AB - BACKGROUND/PURPOSE: As long as the survival rate of patients with abdominal wall defects (AWD) increases, information about long-term follow-up is becoming necessary. Even though quality of life in these patients, in absence of associated anomalies, appears to be unaffected, respiratory impairment soon after birth has been documented; therefore, participation in sports rarely is addressed. METHODS: Eighteen patients, ranging in age from 7 to 18 years, operated on at birth for large abdominal wall defects (> 4 cm for gastroschisis; >6 cm for omphalocele) were asked to come for a stress test on a treadmill, with measurements of time of exercise (TE), maximal oxygen consumption (VO2 max) and continuous recording of vital parameters. Respiratory function also was assessed by Forced Vital Capacity (FVC). RESULTS: Ergometric data were compared with those of a normal pediatric population. All patients were able to perform the stress test with no cardiovascular abnormalities detected at rest or on exertion. Maximum heart rate was reached after a significantly shorter TE, and VO2 max was significantly reduced when comparing normal subjects with AWD subjects and AWD subjects in sports with those sedentary. FVC was only slightly reduced in AWD patients without reaching statistical significance. CONCLUSIONS: These findings indicate that patients operated on for AWD at birth exhibit a normal cardiorespiratory function; decreased TE and VO2 max are likely to reflect a lack of physical activity with a lower degree of fitness. Therefore, no limitations to motor performances should exist for these patients. Well-being may be greatly improved by regular exercise. PMID- 12632370 TI - Fibroblast growth factor signaling in the developing tracheoesophageal fistula. AB - BACKGROUND/PURPOSE: The Adriamycin-induced rat model of esophageal atresia and tracheoesophageal fistula (EA/TEF) provides a reliable system for the study of EA/TEF pathogenesis. The authors previously hypothesized that faulty branching lung morphogenesis pathways were a critical component of its pathogenesis. The authors have found evidence for faulty fibroblast growth factor (FGF) signaling related to epithelial-mesenchymal interactions in the fistula tract. To better define FGF signaling, the differential expression of FGF ligands and their receptors between lung, fistula tract, and esophagus are described. METHODS: Time dated pregnant, Sprague-Dawley rats were injected with Adriamycin (2 mg/kg intraperitoneally) on days 6 through 9 of gestation. Tissues were processed for histology and reverse transcriptase polymerase chain reaction. FGF-1, -7 and -10 were measured from whole lung, fistula tract, and esophagus of TEF or normal embryos. Expression of FGF2RIIIb and FGF2RIIIc receptors was measured in isolated epithelium and mesenchyme of lung and fistula tract of TEF embryos as well as lung and esophagus from normal controls. RESULTS: FGF-1 mRNA was present in the fistula tract and normal and Adriamycin-exposed lung but absent from whole esophagus. Interestingly, FGF-7 mRNA was present only in normal lung. FGF-10 was present in all tissues examined. FGF2RIIIb mRNA was absent in fistula mesenchyme but present in all other tissues examined. However, the splice variant FGF2RIIIc mRNA was present in all tissues examined. CONCLUSIONS: These findings support defective FGF signaling in the rat model of EA/TEF. Absence of FGF-7 mRNA in Adriamycin-exposed tissues suggests the primary effect of Adriamycin may be to inhibit FGF-7 expression. Moreover, absence of FGF2RIIIb in fistula mesenchyme may be caused by loss of positive feedback from FGF-7, its normal obligate ligand. Understanding these specific defects in FGF signaling may provide insight into faulty mechanisms of EA/TEF. PMID- 12632372 TI - Development of a novel method to predict disability after head trauma in children. AB - BACKGROUND/PURPOSE: Although analysis of functional independence measures (FIM) at discharge are useful for assessing extent of disability in head-injured children, there is no reliable method to predict the severity of disability at the time of admission. The authors developed a novel method to predict severe disability after head trauma on admission. METHODS: Head-injured patients, 2 to 16 years old, with FIM recorded at discharge (n = 3,491) were identified in our state trauma database for the period from 1993 through 1996. Patients categorized as completely dependent by one or more of the FIM (Feeding, Locomotion, Expression, Transfer Mobility, Social Interaction) were classified as disabled. Probability of disability (P(D)) was estimated based on regression weights for Glasgow Coma Scale (GCS), Injury Severity Score (ISS), age, and number of anatomic regions injured. Observed to expected disability rates were compared using a test data set of 2,553 patients entered in the database between 1997 through 1999. RESULTS: There was no statistically significant difference between observed and expected disability across all P(D) intervals, which suggests that the P(D) accurately predicted disability. CONCLUSIONS: P(D) offers a novel and reliable method for early prediction of likelihood of disability in children who sustain head trauma. Routine use of the P(D) may lead to earlier intervention to improve long-term results in head-injured children. PMID- 12632373 TI - Differential expression of Bcl-2 and Bax may enhance neuroblastoma survival. AB - BACKGROUND/PURPOSE: Aggressive tumors may alter their expression of Bcl-2 proteins to decrease apoptosis and increase survival. The authors reported previously that neuroblastoma cells have diminished apoptosis when placed in coculture with hepatocytes to stimulate a metastatic environment. It was hypothesized that the expression of proapoptotic (Bax) and prosurvival (Bcl-2 and Mcl-1) proteins would be altered in neuroblastoma cells grown in a cell culture model of metastatic neuroblastoma. METHODS: Human neuroblastoma cells (IMR-32) were grown alone or in coculture with human hepatocytes for 2, 3, or 4 days. Bcl 2, Mcl-1, and Bax mRNA were measured with reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Bcl-2, an antiapoptotic protein, was significantly increased in cocultured neuroblastoma cells by day 4. Bax, a proapoptotic protein, was significantly diminished by day 3. No significant change in Mcl-1 occurred in this study. CONCLUSIONS: When neuroblastoma cells placed in coculture, the prosurvival protein, Bcl-2, is upregulated whereas the proapoptotic protein, Bax, is downregulated. The combination of these changes can maximally enhance the survival rate of neuroblastoma cells in coculture. The propensity for neuroblastoma to either metastasize or regress may be associated with its ability to differentially regulate the expression of different members of the Bcl-2 protein family. PMID- 12632375 TI - Mutation analysis of the RET gene in total intestinal aganglionosis by wave DNA fragment analysis system. AB - BACKGROUND/PURPOSE: Total intestinal aganglionosis (TIA) extending from the duodenum to the rectum is the most rare form of Hirschprung's disease (HSCR) and usually is fatal. RET is the major gene associated with HSCR, and germline mutations of this gene account for up 50% of familial and up to 15 to 20% of sporadic cases in HSCR. The aim of this study was to investigate DNA variants in the RET gene in TIA patients using the WAVE DNA Fragment Analysis System. METHODS: Genomic DNA was extracted from whole blood samples from 6 patients with TIA. Polymerase chain reaction (PCR) amplification of the 21 exons of RET was performed using published oligonucleotide primers. Heteroduplexes were followed by the WAVE DNA Fragment Analysis System with the DNASep cartridge. RESULTS: WAVE system technology detected 16 variants in the RET gene in the 6 patients with TIA. Three patients had a significant mutation in exon 8, 11, and 15, respectively. Thirteen RET polymorphic variants also were detected in the 6 patients, with L746L variant in exon 13 occurring in 4 patients. CONCLUSIONS: WAVE system technology is an efficient method for the detection of DNA sequence variants. Our findings suggest that not only RET mutations but also RET polymorphic variants may contribute to the occurrence of TIA. PMID- 12632374 TI - Patent ductus arteriosus in micropreemies and full-term infants: the relative merits of surgical ligation versus indomethacin treatment. AB - BACKGROUND/PURPOSE: Treatment of ductus arteriosus is controversial. The merits and timing of ductus ligation versus indomethacin treatment were compared. METHODS: Study parameters for infants with PDA (June 1988 through March 2001) included age, weight, physical findings, echocardiograph, success/complications of treatment, and length of stay (LOS). Statistical analysis was by descriptive univariate analysis. RESULTS: Two hundred twelve infants were included. Median gestational age was 26 weeks (range, 22-38); weight, 836 g (447 to 2,863). Findings included murmur (94%), congestion (20%), cardiomegaly (15%), bounding pulses (6%), hyperdynamic precordium (4%), and CHF (2%). Echocardiographic measurements were left atrial diameter, 0.92 cm; posterior wall thickness, 0.26 cm; PDA diameter, 2.5 mm; septal thickness, 0.27 cm; aortic root diameter, 0.64 cm; ejection fraction, 39%; left ventricular internal diameter-diastole, 1.3 cm; left ventricular internal diameter-systole, 0.82 cm; right ventricular internal diameter-diastole, 0.51 cm. No measurement, except PDA diameter, was predictive of medical failure or need for reoperation. However, weight less than 1,000 g was highly predictive of medical failure. Additional cardiac anomalies included ASD (69%), VSD (3%), and aortic coarctation (1%). One hundred sixty-seven babies (79%) received indomethacin, closing the PDA in 88 cases (53%, recurrence = 47%). Indomethacin complications (73%) included thrombocytopenia (36%), BUN increase (31%), sepsis (30%), oliguria (25%), hyponatremia (25%), IVH (16%), pulmonary interstitial emphysema (11%), NEC (8%), intestinal perforation (4%), and bleeding (3%). Seventy-seven babies (36%) required operations (92% in the NICU) performed by pediatric surgeons. Ligation criteria included hypoxia, hypercapnia, decreasing compliance, CHF, and contraindications/failure of indomethacin. Complications included pneumothorax (4%), IVH (4%), bleeding (4%), NEC (1%), and wound infection (1%). LOS averaged 82 days. CONCLUSIONS: Although indomethacin therapy is a reasonable treatment alternative, it is associated with significant complications. Ductus ligation may be preferable, especially in very low birth weight babies, because it is associated with low morbidity and almost certain degree of success. PMID- 12632376 TI - Ileal resection enhances jejunal absorptive adaptation for water and electrolytes to extrinsic denervation: implications for segmental small bowel transplantation. AB - BACKGROUND/PURPOSE: Segmental small bowel transplantation (SBT) may be an alternative to whole jejunoileal SBT. The aim of this study was to evaluate adaptability of extrinsically denervated jejunum after ileectomy. METHODS: Baseline absorption of an isomolar, nonnutrient electrolyte solution was measured in an 80-cm in situ jejunal segment. Control (CON) and extrinsically denervated dogs (EXT DEN) then underwent distal 50% enterectomy; EXT DEN dogs also underwent a complete extrinsic denervation of remnant jejunum. Absorption was remeasured 2 and 12 weeks later; jejunal biopsies at 12 weeks were compared with baseline. RESULTS: All dogs experienced weight loss and diarrhea, which resolved by 10 weeks in CON but persisted in EXT DEN dogs. Net absorptive fluxes of water, Na+, K+, and Cl-, however, remained unchanged in both groups 2 weeks and 12 weeks after denervation. Morphometric evaluation showed an increase in crypt depth in both groups at the 12-week time-point. CONCLUSIONS: Despite a clinical short bowel syndrome, more weight loss, and diarrhea in EXT DEN, there were no temporal differences in net absorptive fluxes of water and electrolytes within or between these 2 groups, and both developed increased crypt depth at 12 weeks. Extrinsic denervation does not blunt jejunal adaptive response to ileectomy. PMID- 12632377 TI - Resolution of hydrops fetalis in congenital cystic adenomatoid malformation after prenatal steroid therapy. AB - BACKGROUND/PURPOSE: Development of hydrops fetalis in fetuses with congenital cystic adenomatoid malformations (CCAM) is a significant risk factor for fetal or neonatal demise. In rare cases, resolution of CCAM has occurred, presumably owing to lesion maturation or involution. Steroid therapy, utilized for lung maturity, has been postulated to accelerate this process. The natural history of hydropic fetuses with CCAM after receiving steroid therapy is presented. METHODS: The authors prospectively followed up with 3 patients who had antenatally diagnosed CCAM and nonimmune hydrops fetalis with predicted mortality. All patients declined or were not candidates for fetal intervention and were treated with standard prenatal betamethasone to increase lung maturity. RESULTS: Three fetuses had CCAM and nonimmune hydrops fetalis diagnosed prenatally. After a course of prenatal steroids during the second trimester, all 3 patients had resolution of their hydrops and were delivered at term without respiratory distress. CONCLUSIONS: Nonimmune hydrops fetalis in fetuses with congenital cystic adenomatoid malformation is a harbinger for fetal demise. The resolution of hydrops in these patients after receiving steroid therapy is an interesting and compelling observation. Because the mechanism of this process is speculative, further studies are needed to elucidate the relationship between antenatal steroids and maturation of congenital cystic adenomatoid malformation. PMID- 12632378 TI - Clostridium botulinum toxin inhibits myometrial activity in vitro: possible application on the prevention of preterm labor after fetal surgery. AB - BACKGROUND/PURPOSE: The current study aimed to analyze the effects of Clostridium botulinum toxin (Botox) on pregnant myometrium activity in vitro. METHODS: Strips of myometrium were obtained from pregnant Wistar rats on gestational day 13 through 15 and placed under controlled conditions within tissue baths containing DeJalon solution. Muscular activity, including amplitude and frequency of contractions, was recorded by a force transducer connected to a polygraph. After stable baseline values were recorded, different concentrations of Botox were added to the tissue baths. Myometrial activity data points for each drug concentration were entered as mean percentual variations of the baseline. A total of 26 uterine samples from 13 animals were studied. Statistical analysis was by single-factor analysis of variance (ANOVA) with P <.05 considered significant. RESULTS: Except for a narrow concentration range, when the effects were nonmonotonic, both amplitude and frequency of myometrial contractions were significantly depressed (P <.05) and eventually totally abolished at most concentrations studied, albeit in a potentially biphasic pattern. Those effects could be reversed by a complete washout of the tissue bath. CONCLUSIONS: Within appropriate concentrations, Botox consistently inhibits or completely arrests myometrial activity in potentially reversible fashion. This agent may prove valuable in premature labor prevention after fetal surgery. PMID- 12632379 TI - VEGF receptors are differentially expressed by neuroblastoma cells in culture. AB - BACKGROUND/PURPOSE: Vascular endothelial growth factor (VEGF) is best known for its angiogenic properties, but its mitogenic capacity may be more important for tumorigenesis. The ability of VEGF to induce specific biologic activities may be dependent on the amount and type of VEGF receptors present. The authors hypothesize that neuroblastoma cells express specific VEGF receptors and that their expression may be altered when the cells are exposed to differing cytokines and culture environments. METHODS: Four groups of human neuroblastoma cells (IMR 32) are studied. (1) Control cells: cultured in standard media. (2) VEGF cells: VEGF added to the media. (3) Tumor necrosis factor alpha (TNF-alpha) cells: TNF alpha added to the media. (4) Serum starved cells: cultured in serum-depleted media. Reverse transcriptase polymerase chain reaction (RT-PCR) is utilized to measure the VEGF receptors flt-1, KDR/flk-1, flt-4, neuropilin 1 (NRP-1), and neuropilin 2 (NRP-2). RESULTS: Flt-1 and KDR are not detected in any groups. Flt 4, NRP-1, and NRP-2 are present in the IMR-32 cells, and their expression is significantly increased by the administration of VEGF. Neuroblastoma cells cultured with TNF-alpha or in serum-depleted media have a significant decrease in the expression of these receptors. CONCLUSIONS: The authors show that neuroblastoma cells express specific VEGF receptors that may be altered by mitogenic or apoptotic stimuli. Specifically targeting VEGF and its receptors may be another therapeutic strategy for the treatment of neuroblastoma. PMID- 12632380 TI - Bilateral congenital diaphragmatic hernia. AB - BACKGROUND/PURPOSE: CDH occurs in approximately 1 in 2,450 live births. Bilateral CDH, previously identified through a limited number of case reports, is extremely rare. The care of CDH patients is a challenge for neonatologists and surgeons. This report details the management and outcome of patients with bilateral CDH. METHODS: Records of all liveborn patients with CDH between 1995 and 2001 in 83 hospitals were entered into the CDH database. Those with bilateral CDH were reviewed retrospectively. Data were analyzed using the Chi;(2) test. RESULTS: A total of 1833 patients were entered in the database, 17 of these had bilateral CDH (0.9%). Eleven were boys. The average birth weight was 2.6 kg. The average gestational age was 36.8 weeks. Sixteen patients experienced early distress requiring intubation (12 immediately), and 4 were placed on extracorporeal membrane oxygenation (ECMO). Seven patients were diagnosed prenatally. Twelve patients (70%) were found to have other anomalies, 3 had chromosomal abnormalities and 7 had cardiac anomalies. These included tetralogy of Fallot (TOF), VSD, absence of the pericardium, coarctation of the aorta (2), accessory SVC with aortic coarctation, and ASD with TOF. Only 9 of 17 (53%) patients underwent surgical repair (6 primary, 3 patch). Mortality rate was 65% compared with 33% of patients with unilateral CDH (P = 0.01). Seven patients died within 48 hours of birth. There was no significant difference in survival based on gender, weight, gestational age, presence of anomalies, or prenatal diagnosis. CONCLUSIONS: The management of infants with bilateral congenital diaphragmatic hernia remains a difficult problem with a significant mortality. Bilateral congenital diaphragmatic hernia is associated more frequently with other major anomalies than unilateral congenital diaphragmatic hernia and should prompt an evaluation for further anomalies. PMID- 12632382 TI - Perfluorohexyloctane (F6H8) induces structural modifications and increases apoptosis in rat primary retinal cultures. AB - The effects of perfluorohexyloctane (F6H8), recently investigated as a long-term artificial vitreous substitute, were studied in vitro, with the use of rat retinal cultures seeded on microporous inserts that allow the cell layer to be in contact with the material to be tested, on the apical side, and with the nutrient medium, on the basal side. After 72 h of treatment with F6H8, retinal cultures lost the characteristic two-layered organization with glial cells at the bottom and neuronal cells on top of them. They appeared to be composed of only one layer of polyhedrical, flattened, and disconnected cells. TUNEL assay revealed an evident increase in the percentage of apoptotic cells in F6H8-treated cultures (30.1 +/- 4.5), compared to control (10.3 +/- 2.6) and perfluoroctane-treated cultures (10.1 +/- 1.7). Immunolabeling of MAP-2, a protein of neuronal cytoskeleton, evidenced a marked loss of neurites. The results suggest that F6H8 is harmful to retinal cells in vitro and can therefore be potentially noxious to the retina as an artificial vitreous substitute. PMID- 12632383 TI - Tobramycin and gentamycin elution analysis between two in situ polymerizable orthopedic composites. AB - This research analyzed Tobramycin and Gentamycin elution characteristics for two antibiotic-impregnated bone composites: PMMA-based Simplex P and the novel, hybrid, bioactive, CORTOSS. Experimental results were correlated with composite hydrophilicity and antibiotic phase partitioning behaviors. The phase partitioning experiment was conducted to understand antibiotic solubility in aqueous environments. By comparing experimental results with calculated data, antibiotic release behavior was predicted. Total Tobramycin elution percentages from CORTOSS and Simplex P were 12.5 and 6.4%, respectively. Total Gentamycin elution percentages from CORTOSS and Simplex P were 6.95 and 10.17%, respectively. Phase partitioning data indicate 100% of Tobramycin remains in aqueous phases, being extremely hydrophilic. This is supported by its calculated theoretical value (log P = - 7.32). Results suggest that Tobramycin elution can be attributed to composite hydrophilicity as well as its high degree of hydrophilicity. Fifteen percent of Gentamycin distributes in hydrophobic phases (log P = - 4.22). Despite a lower Gentamycin hydrophilicity, its release was affected by its complexation with polar salts in the leaching buffer, thereby increasing its elution potential, making it appreciably water soluble. CORTOSS is more hydrophilic; therefore the migration of aqueous liquids into the polymer network of CORTOSS facilitates greater antibiotic elution compared with hydrophobic Simplex P. PMID- 12632384 TI - Cell proliferation and differentiation during fracture healing are influenced by locally applied IGF-I and TGF-beta1: comparison of two proliferation markers, PCNA and BrdU. AB - Growth factors IGF-I and TGF-beta1 are known to stimulate fracture healing. The purpose of this study was to investigate the role of locally applied IGF-I and TGF-beta1 during the early phase of fracture healing (Days 5, 10, and 15 after fracture) on cellular processes like proliferation and differentiation in a rat model. Two different immunohistochemical markers were used to analyze cell proliferation: (1) injection of the thymidine analogue BrdU and subsequent immunohistochemical staining for BrdU-positive nuclei, and (2) the antibody against the "proliferating cell nuclear antigen" (PCNA). In comparison, both methods revealed similar results concerning the types of proliferating cells at the different time points and the two groups. Labeling indices of both methods showed very good correlation (e.g., r(s): 0.887 and p < 0.001 at day 10 in the control group without growth factors). Comparison of the callus morphology and the proliferation rate showed differences during fracture healing due to the local application of IGF-I and TGF-beta1 from coated implants. At Day 5 the callus of the group treated with growth factors displayed an earlier appearance of cartilage compared to the control group. This was accompanied by an onset of cell proliferation in chondrocytes. Likewise, at the later time points an enhanced maturation of the callus tissue and the proliferation pattern were detectable in the growth-factor group. These results indicate that local application of IGF-I and TGF-beta1 accelerates early cellular processes during fracture healing. PMID- 12632385 TI - IGF-I and TGF-beta 1 incorporated in a poly(D,L-lactide) implant coating stimulates osteoblast differentiation and collagen-1 production but reduces osteoblast proliferation in cell culture. AB - Previous in vivo studies revealed a stimulating effect of locally applied IGF-I and TGF-beta1 released from poly(D,L-lactide)-coated titanium implants on rat and porcine fracture healing. The purpose of the present study was to evaluate the effect of IGF-I (5% w/w) and TGF-beta1 (1% w/w) and the carrier PDLLA on osteoblasts in cell culture to improve the understanding of these growth factors. The well-characterized human osteoblast cell line hFOB 1.19 was used in the study. The implants and cells were cocultured in a noncontact manner. The cells were incubated for 10 days in total, and the implants (n = 6 each group and time point) were added for 1 h, 12 h, 24 h, 2 d, 4 d, or 10 d. To analyze a possible effect of the growth factors or the coating, cell proliferation, metabolism, and differentiation were investigated. As an indicator for differentiation the production of collagen I was chosen. All experimental groups showed comparable cell vitality. No change in the pH of the medium was detectable between the analyzed groups. When the effect of the titanium implant and the PDLLA coating were compared with the control culture, no differences in proliferation, metabolic activity, and collagen I production were detectable. The osteoblasts treated with IGF-I and TGF-beta1 released from PDLLA revealed a significantly enhanced collagen I production with a decrease in proliferation and metabolic activity compared to the other groups. No significant differences in collagen I production were seen due to the incubation time points. None of the experimental groups evoked an immunological response on mouse macrophages. In conclusion, the PDLLA-carrier showed no negative effect on osteoblasts, whereas the incorporated growth factors stimulated osteoblast differentiation. PMID- 12632386 TI - Molecular biologic comparison of new bone formation and resorption on microrough and smooth bioactive glass microspheres. AB - In a recent in vitro study, chemical microroughening of a bioactive glass surface was shown to enhance attachment of MG-63 osteoblastic cells to glass. The current study was designed to delineate the effects of microroughening on the gene expression patterns of bone markers during osteogenesis and new bone remodeling on bioactive glass surface in vivo. With the use of a rat model of paired comparison, a portion of the medullary canal in the proximal tibia was evacuated through cortical windows and filled with microroughened or smooth bioactive glass microspheres. The primary bone-healing response and subsequent remodeling were analyzed at 1, 2, and 8 weeks, respectively, by radiography, pQCT, histomorphometry, BEI-SEM, and molecular biologic analyses. The expression of various genes for bone matrix components (type I collagen, osteocalcin, osteopontin, osteonectin) and proteolytic enzymes (cathepsin K, MMP-9) were determined by Northern analysis of the respective mRNAs. Paired comparison showed significant differences in the mRNAs levels for specific bone matrix components at 2 weeks: osteopontin was significantly higher (p =.01) and osteonectin significantly lower (p =.05) in bones filled with microroughened microspheres than in those filled with smooth microspheres. Bones filled with microrough microspheres also showed significantly increased ratios of cathepsin K and MMP-9 (both markers of osteoclastic resorption) to type I collagen (p =.02 and p =.02, respectively) at 2 weeks and a significantly increased expression of MMP-9 at 8 weeks (p =.05). The pQCT, histomorphometric, and BEI-SEM analyses revealed no significant differences in the pattern of bone-healing response. Based on these results, microroughening of a bioactive glass surface could trigger temporal changes in the expression of specific genes especially by promoting the resorption part of new bone-remodeling processes. Future studies are needed to evaluate if the observed changes of gene expression are directly related to the microrough surface of any biomaterial or are biomaterial specific. PMID- 12632387 TI - Rapid endothelialization of PhotoFix natural biomaterial vascular grafts. AB - To date, no off-the-shelf graft has performed better than the autologous vessel in applications requiring small-bore (< 6-mm diameter) vascular grafts. Much research has been devoted to seeding endothelial cells on synthetic grafts to improve their long-term clinical performance. One key challenge is the ability to retain the endothelium on the graft lumen for extended times. The goal of this research was to develop a process to seed endothelial cells inside a vascular graft and to quickly condition the cells so as to minimize their damage or removal under physiological flow. In addition, the use of PhotoFix(R) natural biomaterial grafts as an improved substrate for human umbilical vein endothelial cells has been evaluated. A motorized system that provides uniform cell seeding of a small-diameter graft (4-mm inner diameter, 10-cm length) by automated radial rotation has been developed. The same system is subsequently adapted for gradual increases in flow rates to strengthen the endothelium, which ultimately was exposed to a final flow rate of 300 ml/min. This process is accomplished without graft transfer, decreasing risks of contamination and physical damage. Cell coverage and cell morphology were evaluated with the use of fluorescence microscopy and scanning-electron microscopy to determine the effectiveness of the flow conditioning process. It was found that endothelial cells exhibit roughly 20 50% improved adhesion to PhotoFix vessels compared to fibrin-treated polytetrafluoroethylene (PTFE) synthetic grafts. Flow conditioning for 6 h enhanced in vitro cell retention by 24% and 40% on PhotoFix and PTFE grafts, respectively. PMID- 12632388 TI - Toothbrushing causes elemental release from dental casting alloys over extended intervals. AB - The release of elements from dental alloys has been linked to alloy biocompatibility. Much of the research measuring elemental release has been done in vitro under passive conditions. The current study supplements a previous report that measured elemental release from dental alloys during and after the equivalent of 1 week of toothbrushing. In the current study, toothbrushing times were extended to the equivalent of 2 years, and elemental release was measured during and after brushing, with and without toothpaste. The results showed that for the major classes of dental alloys, brushing alone caused no significant elemental release during the brushing, and only minor increases after brushing. Brushing with toothpaste caused significant increases in elemental release for all elements of all alloys, but the largest increases were for the two nickel based alloys. Nickel released during brushing with toothpaste reached 600-800 microg/cm(2) of alloy surface. Both beryllium-containing and non-beryllium containing nickel-based alloys behaved similarly, refuting claims that non beryllium alloys are superior in this regard. Thus, brushing with toothpaste under these extended in vitro conditions appears to increase the biological liabilities from elemental release for all alloys, but primarily for nickel-based alloys. PMID- 12632389 TI - Effect of extent of motion and type of load on the wear of polyethylene in a biaxial hip simulator. AB - The most commonly used wear test device for prosthetic hip joints is the so called biaxial rocking motion (BRM) design. The design has been criticized for its excessive sliding distance per cycle. The design was modified so that the extent of motion was reduced from 46 to 23 degrees, and comparative tests were run with the use of 1-kN static load. The present authors have earlier used 1-kN static load with good results. To further confirm this finding, additional, comparative tests were run with double-peak dynamic load and 46 degrees extent of motion. All three tests (46 degrees /static, 23 degrees /static and 46 degrees /dynamic), were done with conventional ultra-high-molecular-weight polyethylene acetabular cups against polished CoCr femoral heads in diluted calf serum lubricant. In all tests, the principal cup wear mechanism was adhesive, manifested as burnishing. With respect to wear particles, those produced in the 46 degrees /dynamic test showed the lowest resemblance to particles isolated from periprosthetic tissues. The 46 degrees /dynamic test produced a mean wear rate 43% higher than 46 degrees /static, whereas the 46 degrees /static and 23 degrees /static tests produced almost identical values. The results indicated that the 46 degrees extent of motion and dynamic load may not always be the optimal combination in BRM tests. PMID- 12632390 TI - Surface conditions of Nitinol wires, tubing, and as-cast alloys. The effect of chemical etching, aging in boiling water, and heat treatment. AB - The surface conditions of Nitinol wires and tubing were evaluated with the use of X-ray photoelectron spectroscopy, high-resolution Auger spectroscopy, electron backscattering, and scanning-electron microscopy. Samples were studied in the as received state as well as after chemical etching, aging in boiling water, and heat treatment, and compared to a mechanically polished 600-grit-finish Nitinol surface treated similarly. General regularities in surface behavior induced by the examined surface treatments are similar for wires, tubing, and studied as cast alloy, though certain differences in surface Ni concentration were observed. Nitinol wires and tubing from various suppliers demonstrated great variability in Ni surface concentration (0.5-15 at.%) and Ti/Ni ratio (0.4-35). The wires in the as-received state, with the exception of those with a black oxide originating in the processing procedure, revealed nickel and titanium on the surface in both elemental and oxidized states, indicating a nonpassive surface. Shape-setting heat treatment at 500 degrees C for 15 min resulted in tremendous increase in the surface Ni concentration and complete Ni oxidation. Preliminary chemical etching and boiling in water successfully prevented surface enrichment in Ni, initially resulting from heat treatment. A stoichiometric uniformly amorphous TiO(2) oxide generated during chemical etching and aging in boiling water was reconstructed at 700 degrees C, revealing rutile structure. PMID- 12632391 TI - Properties and performance of sulfide-substituted plasticized poly(vinyl chloride) as a biomaterial. AB - Plasticized poly(vinyl chloride) (PVC) was surface modified by nucleophilic substitution of the chlorine atoms of PVC by sulfide ions in aqueous media in the presence of a phase-transfer catalyst (PTC) to make it migration resistant. The modified PVC was sterilized by steam autoclaving and gamma radiation and subjected to plasticizer extraction in an extraction medium such as hexane. Virtually no plasticizer migrated out of modified PVC into the medium even after prolonged incubation for 30 d. The properties of the modified surface were evaluated by contact angle measurements, scanning electron microscopy (SEM), cell culture studies, hemolysis assay, and whole-blood clotting time measurements. The mechanical properties of modified sheets were evaluated by measurements of tensile strength and strain at break after subjecting the specimens to different modes of sterilization. The modified PVC surface showed enhanced wettability and was found to be non-hemolytic and non-cytotoxic. Whole-blood clotting profiles of unmodified and modified PVC were virtually the same. Modification imparted slight yellow color to the material. Surface modification resulted in an 8% decrease in the ultimate stress and 28% decrease in the strain at break values. The sulfide substituted PVC was found to be blood compatible and would be useful in applications such as tubing for transfusion, dialysis etc., where migration resistance is important. PMID- 12632392 TI - Composition and in vitro biocompatibility of corroding tungsten coils. AB - The purpose of the article is to evaluate composition and biocompatibility of corroding mechanically detachable spirals (MDS, Balt Extrusion, Montmorency, France). Analysis of the material composition of corroding MDS coils was assessed by inductively coupled plasma atomic emission spectroscopy, inductively coupled plasma mass spectroscopy, and wavelength-dispersive x-ray spectrometry. Toxicity assays were performed with human venous endothelial cells, venous smooth muscle cells and fibroblasts. The analyses of the MDS coils demonstrated a tungsten content of the dissolving MDS spirals of > 99.9 mas%. In vitro, human endothelial, vascular smooth muscle cells and fibroblasts were not adversely affected by markedly elevated tungsten concentrations (60,500 microg/l) after 12 days in the culture medium. The examined cells showed an extensive vital growth on the coil surface. Corrosion of tungsten coils leads to markedly elevated tungsten levels in the culture medium. However, growth and vitality of endothelial cells, fibroblasts, and vascular smooth muscle cells are not adversely affected by elevated tungsten concentrations. PMID- 12632393 TI - Simple and efficient solid support scavenging of excess acyl donors after enzymatic acylations in organic solvents. AB - A simple and efficient method for removing excess acyl donors following enzymatic acylations in organic solvents was developed. This method is based on selective chemical scavenging of acyl donors using an amino-functionalized solid support, and does not affect the desired acylated product. A wide variety of different acyl donors, including vinyl and trifluoroethyl esters and vinyl carbonates, can be quantitatively removed by this method, thus providing a simple and highly efficient tool for purification of reaction products after enzymatic acylation. PMID- 12632394 TI - Visualization of alginate-poly-L-lysine-alginate microcapsules by confocal laser scanning microscopy. AB - Confocal laser scanning microscopy (CLSM) was used to study the distribution of polymers and cross-linking ions in alginate-poly-L-lysine (PLL) -alginate microcapsules made by fluorescent-labeled polymers. CLSM studies of Ca-alginate gel beads made in the presence and absence of non-gelling sodium ions revealed a more inhomogeneous distribution of alginate in beads formed in the absence of non gelling ions. In the formation of alginate-PLL capsules, the polymer gradients in the preformed gel core were destabilized by the presence of non-gelling ions in the washing step and in the PLL solution. Ca-alginate gels preserved the inhomogeneous structure by exposure to ion-free solution in contrast to exposure to non-gelling ions (Na(+)). By exchanging Ca(2+) with Ba(2+) (10 mM), extremely inhomogeneous gel beads were formed that preserved their structure during the washing and exposure to PLL in saline. PLL was shown to bind at the very surface of the alginate core, forming a shell-like membrane. The thickness of the PLL layer increased about 100% after 2 weeks of storage, but no further increase was seen after 2 years of storage. The coating alginate was shown to overlap the PLL layer. No difference in binding could be observed among coating alginates of different composition. This paper shows an easy and novel method to study the distribution of alginate and PLL in intact microcapsules. As the labeling procedures are easy to perform, the method can also be used for a variety of other polymers in other microencapsulation systems. PMID- 12632395 TI - Determination of laccase activity in mixed solvents: comparison between two chromogens in a spectrophotometric assay. AB - Spectrophotometric determination of laccase activity with ABTS acting as chromogen yields exceedingly low values whenever conducted in a water-organic mixed solvent. Nevertheless, there is firm evidence that laccase is able to oxidize substrates such as phenols and amines quantitatively in these mixed solvents. We show that the apparently small rate of ABTS oxidation by laccase in a mixed solvent, such as buffered water-dioxane 1:1, is not amenable to the denaturation of laccase but rather to the decreased stability of ABTS(.+). We propose HAA as a more reliable chromogen for the determination of laccase activity in mixed solvents. PMID- 12632396 TI - Laser impact assessment in a biofilm-forming bacterium Pseudoalteromonas carrageenovora using a flow cytometric system. AB - Impact by pulsed laser irradiations from an Nd:YAG laser on the marine biofilm forming bacterium Pseudoalteromonas carrageenovora has been studied using a flow cytometric system. The biofilm-forming bacteria in the planktonic state have been irradiated while flowing, and the mortality and bacterial attachment have been determined by exposing TiN coupons in the system. Coupons suspended in the non irradiated bacterial flow were treated as the control. The fluence used in the study was 0.1 J/cm(2). Three flow rates (14, 28, and 42 cm/min) and two exposure durations (15 and 30 min) were tested. The results showed the increase in bacterial mortality with the decrease in flow rate. The maximum mortality of 27.5% was observed when the flow rate was 14 cm/min. The bacterial attachment increased with the increase in flow rate and exposure duration. The area of bacterial attachment on the experimental coupons exposed to the irradiated sample was significantly lesser than that for the nonirradiated sample. The results thus show in a flowing system, low power pulsed laser irradiations could reduce the bacterial attachment even though it did not cause significant mortality. PMID- 12632397 TI - High-density seeding of myocyte cells for cardiac tissue engineering. AB - Tissue engineering of 1- to 5-mm-thick, functional constructs based on cells that cannot tolerate hypoxia for prolonged time periods (e.g., cardiac myocytes) critically depends on our ability to seed the cells at a high and spatially uniform initial density and to maintain their viability and function. We hypothesized that rapid gel-cell inoculation in conjunction with direct medium perfusion through the seeded scaffold would increase the rate, yield, viability, and uniformity of cell seeding. Two cell types were studied: neonatal rat cardiomyocytes for feasibility studies of seeding and cultivation with direct medium perfusion, and C2C12 cells (a murine myoblast cell line) for detailed seeding studies. Cells were seeded at densities corresponding to those normally present in the adult rat heart ([0.5-1] x 10(8) cells/cm(3)), into collagen sponges (13 mm x 3 mm discs), using Matrigel as a vehicle for rapid cell delivery. Scaffolds inoculated with cell-gel suspension were seeded either in perfused cartridges with alternating medium flow or in orbitally mixed Petri dishes. The effects of seeding time (1.5 or 4.5 h), initial cell number (6 or 12 million cells per scaffold), and seeding set-up (medium perfusion at 0.5 and 1.5 mL/min; orbitally mixed dishes) were investigated using a randomized three-factor factorial experimental design with two or three levels and three replicates. The seeding cell yield was consistently high (over 80%), and it appeared to be determined by the rapid gel inoculation. The decrease in cell viability was markedly lower for perfused cartridges than for orbitally mixed dishes (e.g., 8.8 +/- 0.8% and 56.3 +/- 4%, respectively, for 12 million cells at 4.5 h post seeding). Spatially uniform cell distributions were observed in perfused constructs, whereas cells were mainly located within a thin (100-200 microm) surface layer in dish seeded constructs. Over 7 days of cultivation, medium perfusion maintained the viability and differentiated function of cardiac myocytes, and the constructs contracted synchronously in response to electrical stimulation. Direct perfusion can thus enable seeding of hypoxia-sensitive cells at physiologically high and spatially uniform initial densities and maintain cell viability and function. PMID- 12632398 TI - Halogenated monoterpene production by microplantlets of the marine red alga Ochtodes secundiramea within an airlift photobioreactor under nutrient medium perfusion. AB - Macrophytic marine red algae are a unique source of novel and bioactive terpenoids, including halogenated monoterpenes. Biomass and halogenated monoterpene production by regenerated microplantlet suspension cultures derived from the red alga Ochtodes secundiramea were studied within a perfusion airlift photobioreactor. Photobioreactor cultivations were carried out at 26 degrees C, 140 microE m(-2)s(-1) light intensity, 0.3 air L(-1) culture min(-1) aeration (3500 ppm CO(2)), and ESS/seawater medium perfusion rate of 0.2 L medium L(-1) culture d(-1). Macronutrient concentrations in the perfusion medium were adjusted to provide nitrate delivery rates of 0.0063, 0.077, and 0.74 mmol L(-1) d(-1) at a fixed N:P ratio of 19:1. Growth was maximized at the highest nutrient delivery rate, where 10 g dry biomass L(-1) culture was achieved after 30 days of cultivation. GC-MS analysis of dichloromethane extracts from cell biomass revealed that O. secundiramea microplantlets produced myrcene, three acyclic halogenated monoterpenes (10-bromomyrcene, 10-bromo-7-chloromyrcene, 3,10 dibromomyrcene), and one cyclic halogenated monoterpene (6-bromo-1,2,8-trichloro 3,4-ochtodene). 10E-bromomyrcene levels were much higher than those of its isomer 10Z-bromomyrcene, demonstrating stereoselective halogenation. Maximum yields of 10E-bromomyrcene and 6-bromo-1,2,8-trichloro-3,4-ochtodene were 15 and 13 micromol/g dry cell mass, respectively. Increasing the rate of nutrient delivery increased the accumulation of myrcene and 10-bromomyrcene during the first 14 days in culture. Furthermore, the yield selectivity toward higher halogenated monoterpenes increased as the rate of nutrient delivery decreased. From this data, a biogenic scheme was proposed where cyclic and acyclic halogenated monoterpenes are derived from sequential halogenation of myrcene, their common precursor. PMID- 12632399 TI - Microfiltration and ultrafiltration of polysaccharides produced by fermentation using a rotating disk dynamic filtration system. AB - The recovery of exopolysaccharides (EPS) produced by Sinorhizobium meliloti bacteria by dynamic microfiltration was investigated using a rotating disk device designed in our laboratory, equipped with a 0.2 microm nylon membrane. This system differs from commercially available systems by the presence of vanes on the disk which produce a very important increase in permeate flux while yielding excellent EPS transmission. For polymers produced under standard fermentation conditions (70 h at 30 degrees C), the mass flux rose to 650 g h(-1) m(-2) using a disk equipped with 2 mm vanes rotating at 2000 rpm against 380 g h(-1) m(-2) with a smooth disk at the same speed. The maximum flux observed was 1560 g h(-1) m(-2) with a 6-mm vanes disk rotating at 3000 rpm and a 36 degrees C broth. An interesting finding was that the permeate flux J(f) for various disks can be correlated by the same function of the mean shear stress at the membrane tau(wm) according to J(f) = 4.6 tau(wm) (0.717) for a 30 degrees C broth, showing that the effect of vanes is merely to increase the shear stress by raising the fluid core velocity between the membrane and the disk. With 6-mm vanes the core angular velocity was found to be 84% of disk velocity vs. 45% for a smooth disk. When the fermentation temperature was increased to 36 degrees C to produce a lower molecular weight polymer, the permeate flux rose by about 250%, much more than what could be expected from the reduction in permeate viscosity and followed the same power law with membrane shear stress as for 30 degrees C. The same device was equipped with a PES 50 kDa membrane to concentrate EPS by ultrafiltration. Permeate fluxes were of the order of 160 L h(-1) m(-2) at 2000 rpm and 30 degrees C with nearly complete EPS rejection. Finally, the net electrical power consumed by the disk was measured by subtracting the power consumed without fluid from the power during filtration at the same speed. This power increases with speed and with the presence of vanes, but since the gain provided by the vanes is very high, the specific energy per m(3) of permeate is minimal with the highest vanes tested (6 mm) and maximal for smooth disks. PMID- 12632400 TI - Effect of methanol feeding strategies on production and yield of recombinant mouse endostatin from Pichia pastoris. AB - Pichia pastoris, a methylotrophic yeast, is an efficient producer of recombinant proteins in which the heterologous gene is under the control of the methanol induced AOX1 promoter. Hence, the accepted production procedure has two phases: In the first phase, the yeast utilizes glycerol and biomass is accumulated; in the second phase, the yeast utilizes methanol which is used both as an inducer for the expression of the recombinant protein and as a carbon source. Since the yeast is sensitive to methanol concentration, the methanol is supplied gradually to the growing culture. Three methanol addition strategies were evaluated for the purpose of optimizing recombinant endostatin production. Two strategies were based on the yeast metabolism; one responding to the methanol consumption using a methanol sensor, and the other responding to the oxygen consumption. In these two strategies, the methanol supply is unlimited. The third strategy was based on a predetermined exponential feeding rate, controling the growth rate at 0.02 h(-1), in this strategy the methanol supply is limited. Throughout the induction phase glycerol, in addition to methanol, was continuously added at a rate of 1 g L h( 1). Total endostatin production was similar in all three strategies, (400 mg was obtained from 3 L initial volume), but the amount of methanol added and the biomass produced were lower in the predetermined rate method. This caused the specific production of endostatin per biomass and per methanol to be 2 times higher in the predetermined rate than in the other two methods, making the growth control strategy not only more efficient but also more convenient for downstream processing. PMID- 12632401 TI - Glucose-6-phosphate dehydrogenase partitioning in two-phase aqueous mixed (nonionic/cationic) micellar systems. AB - The enzyme glucose-6-phosphate dehydrogenase (G6PD) plays an important role in maintaining the level of NADPH and in producing pentose phosphates for nucleotide biosynthesis. It is also of great value as an analytical reagent, being used in various quantitative assays. In searching for new strategies to purify this enzyme, the partitioning of G6PD in two-phase aqueous mixed (nonionic/cationic) micellar systems was investigated both experimentally and theoretically. Our results indicate that the use of a two-phase aqueous mixed micellar system composed of the nonionic surfactant C(10)E(4) (n-decyl tetra(ethylene oxide)) and the cationic surfactant C(n)TAB (alkyltrimethylammonium bromide, n = 8, 10, or 12) can improve significantly the partitioning behavior of G6PD relative to that obtained in the two-phase aqueous C(10)E(4) micellar system. This improvement can be attributed to electrostatic attractions between the positively charged mixed (nonionic/cationic) micelles and the net negatively charged enzyme G6PD, resulting in the preferential partitioning of G6PD to the top, mixed micelle-rich phase of the two-phase aqueous mixed micellar systems. The effect of varying the cationic surfactant tail length (n = 8, 10, and 12) on the denaturation and partitioning behavior of G6PD in the C(10)E(4) /C(n)TAB/buffer system was investigated. It was found that C(8)TAB is the least denaturing to G6PD, followed by C(10)TAB and C(12)TAB. However, the C(10)E(4)/C(12)TAB/buffer system generated stronger electrostatic attractions with the net negatively charged enzyme G6PD than the C(10)E(4)/C(10)TAB/buffer and the C(10)E(4)/C(8)TAB/buffer systems, when using the same amount of cationic surfactant. Overall, the two-phase aqueous mixed (C(10)E(4)/C(10)TAB) micellar system yielded the highest G6PD partition coefficient of 7.7, with a G6PD yield in the top phase of 71%, providing the optimal balance between the denaturing effect and the electrostatic attractions for the three cationic surfactants examined. A recently developed theoretical framework to predict protein partition coefficients in two-phase aqueous mixed (nonionic/ionic) micellar systems was implemented, and the theoretically predicted G6PD partition coefficients were found to be in reasonable quantitative agreement with the experimentally measured ones. PMID- 12632402 TI - Role of pericellular matrix in development of a mechanically functional neocartilage. AB - The role of the chondrocyte pericellular matrix (PCM) was examined in a three dimensional chondrocyte culture system to determine whether retention of the native pericellular matrix could stimulate collagen and proteoglycan accumulation and also promote the formation of a mechanically functional hyaline-like neocartilage. Porcine chondrocytes and chondrons, consisting of the chondrocyte with its intact pericellular matrix, were maintained in pellet culture for up to 12 weeks. Sulfated glycosaminoclycans and type II collagen were measured biochemically. Immunocytochemistry was used to examine collagen localization as well as cell distribution within the pellets. In addition, the equilibrium compressive moduli of developing pellets were measured to determine whether matrix deposition contributed to the mechanical stiffness of the cartilage constructs. Pellets increased in size and weight over a 6-week period without apparent cell proliferation. Although chondrocytes quickly rebuilt a PCM rich in type VI collagen, chondron pellets accumulated significantly more proteoglycan and type II collagen than did chondrocyte pellets, indicating a greater positive effect of the native PCM. After 5 weeks in chondron pellets, matrix remodeling was evident by microscopy. Cells that had been uniformly distributed throughout the pellets began to cluster between large areas of interterritorial matrix rich in type II collagen. After 12 weeks, clusters were stacked in columns. A rapid increase in compressive strength was observed between 1 and 3 weeks in culture for both chondron and chondrocyte pellets and, by 6 weeks, both had achieved 25% of the equilibrium compressive stiffness of cartilage explants. Retention of the in vivo PCM during chondrocyte isolation promotes the formation of a mechanically functional neocartilage construct, suitable for modeling the responses of articular cartilage to chemical stimuli or mechanical compression. PMID- 12632403 TI - Electrochemical optical waveguide lightmode spectroscopy (EC-OWLS): a pilot study using evanescent-field optical sensing under voltage control to monitor polycationic polymer adsorption onto indium tin oxide (ITO)-coated waveguide chips. AB - A new technique has been developed that combines evanescent-field optical sensing with electrochemical control of surface adsorption processes. This new technique, termed "electrochemical optical waveguide lightmode spectroscopy" (EC-OWLS), proved efficient in monitoring molecular surface adsorption and layer thickness changes of an adsorbed polymer layer examined in situ as a function of potential applied to a waveguide in a pilot study. For optical sensing, a layer of indium tin oxide (ITO) served as both a high-refractive-index waveguide and a conductive electrode. In addition, an electrochemical flow-through fluid cell was provided, which incorporated working, reference, and counter electrodes, and was compatible with the constraints of optical sensing. Poly(L-lysine)-grafted-poly(ethylene glycol) (PLL-g-PEG) served as a model, polycation adsorbate. Adsorption of PLL-g PEG from aqueous buffer solution increased from 125 to 475 ng/cm(2 )along a sigmoidal path as a function of increasing potential between 0 and 1.5 V versus the Ag reference electrode. Upon buffer rinse, adsorption was partially reversible when a potential of >/=0.93 V was maintained on the ITO waveguide. However, reducing the applied potential back to 0 V before rinsing resulted in irreversible polymer adsorption. PLL-g-PEG modified with biotin demonstrated similar adsorption characteristics, but subsequent streptavidin binding was independent of biotin concentration. Applying positive potentials resulted in increased adsorbed mass, presumably due to polymer chain extension and reorganization in the molecular adlayer. PMID- 12632404 TI - Levitation and movement of human tumor cells using a printed circuit board device based on software-controlled dielectrophoresis. AB - In this study we describe an original, efficient, and innovative printed circuit board (PCB) device able to generate dielectrophoresis-based, software-controlled cages that can be moved to any place inside a microchamber. Depending on their dielectrophoretic properties, eukaryotic cells can be "entrapped" in cages and moved under software control. The main conclusion gathered from the experimental data reported is that the PCB device based on dielectrophoresis permits levitation and movement of different tumor cells at different dielectrophoresis conditions. The results presented herein are therefore the basis for experiments aimed at forced interactions or separation of eukaryotic cells using "lab-on-a chip." In fact, because many cages can be controlled at the same time, and two or more cages can be forced to share the same or a different location, it is possible, in principle, either to bring in contact cells of a differing histotype or to separate them. PMID- 12632405 TI - Fusion protein of Vitreoscilla hemoglobin with D-amino acid oxidase enhances activity and stability of biocatalyst in the bioconversion process of cephalosporin C. AB - In this study we constructed an artificial flavohemoprotein by fusing Vitreoscilla hemoglobin (VHb) with D-amino acid oxidase (DAO) of Rhodotorula gracilis to determine whether bacterial hemoglobin can be used as an oxygen donor to immobilized flavoenzyme. This chimeric enzyme significantly enhanced DAO activity and stability in the bioconversion process of cephalosporin C. In a 200 mL bioreactor, the catalytic efficiency of immobilized VHb-DAO against cephalosporin C was 12.5-fold higher than that of immobilized DAO, and the operational stability of the immobilized VHb-DAO was approximately threefold better than that of the immobilized DAO. In the scaled-up bioprocess with a 5-L bioreactor, immobilized VHb-DAO (2500 U/L) resulted in 99% bioconversion of 120 mM cephalosporin C within 60 min at an oxygen flow rate of 0.2 (v/v) x min. Ninety percent of the initial activity of immobilized VHb-DAO could be maintained at up to 50 cycles of the enzymatic reaction without exogenous addition of H(2)O(2) and flavin adenine dinucleotide (FAD). The purity of the final product, glutaryl-7-aminocephalosporanic acid, was confirmed to be 99.77% by high performance liquid chromatography (HPLC) analysis. Relative specificity of immobilized VHb-DAO on D-alpha-aminoadipic acid, a precursor in cephalosporin C biosynthesis, increased twofold, compared with that of immobilized DAO, suggesting that conformational modification of the VHb-DAO fusion protein may be altered in favor of cephalosporin C. PMID- 12632406 TI - Monitoring of a sequencing batch reactor using adaptive multiblock principal component analysis. AB - Multiway principal component analysis (MPCA) for the analysis and monitoring of batch processes has recently been proposed. Although MPCA has found wide applications in batch process monitoring, it assumes that future batches behave in the same way as those used for model identification. In this study, a new monitoring algorithm, adaptive multiblock MPCA, is developed. The method overcomes the problem of changing process conditions by updating the covariance structure recursively. A historical set of operational data of a multiphase batch process was divided into local blocks in such a way that the variables from one phase of a batch run could be blocked in the corresponding blocks. This approach has significant benefits because the latent variable structure can change for each phase during the batch operation. The adaptive multiblock model also allows for easier fault detection and isolation by looking at the relationship between blocks and at smaller meaningful block models, and it therefore helps in the diagnosis of the disturbance. The proposed adaptive multiblock monitoring method is successfully applied to a sequencing batch reactor for biological wastewater treatment. PMID- 12632407 TI - Magnetic resonance imaging in the evaluation of bone damage in rheumatoid arthritis: a more precise image or just a more expensive one? PMID- 12632408 TI - Does activation of the innate immune system contribute to the development of rheumatoid arthritis? PMID- 12632409 TI - Interleukin-17 in rheumatoid arthritis: if T cells were to contribute to inflammation and destruction through synergy. PMID- 12632410 TI - Immunologic intervention in the pathogenesis of osteoarthritis. PMID- 12632411 TI - Leonard Trask: the wonderful invalid: the first American description of ankylosing spondylitis. AB - This report describes the first published clinical account of a case of ankylosing spondylitis in the United States. The patient progressively developed very severe kyphosis with resultant severe disability. The condition apparently started after an accident, and the report has interesting medicolegal implications. PMID- 12632412 TI - Computerized measurement of magnetic resonance imaging erosion volumes in patients with rheumatoid arthritis: a comparison with existing magnetic resonance imaging scoring systems and standard clinical outcome measures. AB - OBJECTIVE: One of the major aims of therapy in rheumatoid arthritis (RA) is to prevent erosive disease and subsequent disability. One of the important goals of therapy assessment must therefore be the accurate measurement of damage progression. We undertook this study to assess the feasibility, reliability, and validity of measuring magnetic resonance imaging (MRI) erosion volumes and synovial volumes in the wrists of RA patients with the use of a semiautomated computerized method. METHODS: Twelve subjects with seropositive RA were chosen to reflect a spectrum of RA severity as determined by the clinical Joint Alignment and Motion (JAM) Scale. MRI of the dominant wrist was performed at the same time of day at baseline and at 48 hours. Images were transferred to a workstation. Erosion volumes and synovial volumes were measured on the coronal images using OSIRIS imaging software. All images were reread in random order at 72 hours by 1 observer. The results were compared with erosion scores and global synovitis scores obtained by the same observer using the Outcome Measures in Rheumatology Clinical Trials MRI RA scoring system. Radiographs were performed at baseline and were read on 2 occasions by 1 observer using the Scott modification of the Larsen method. RESULTS: Total erosion volume per subject ranged from 0 cm(3) to 4.7 cm(3). The total synovial membrane volume per subject ranged from 0.1 mm(3) to 12.1 cm(3). Intraclass correlation coefficients for erosion volumes and synovial volumes demonstrated excellent intraobserver reliability and interoccasion reliability. There was a strong positive correlation between the total erosion volume and the total erosion score. The correlation between the synovial volumes and synovitis score was less favorable (r = 0.53-0.86). Positive correlations were demonstrated between the erosion volumes, the JAM score, and the modified Larsen scores. No significant correlation was demonstrated between the erythrocyte sedimentation rate, C-reactive protein level, swollen joint count, tender joint count, or the Disease Activity Score in 28 swollen and 28 tender joints and the synovial volumes or synovitis scores. CONCLUSION: This study is the first to demonstrate the feasibility, reliability, and validity of computerized MRI erosion volume measurements in the wrists of RA patients. The method will require further evaluation in terms of interobserver reliability, with examination of responsiveness in longitudinal studies, but the method demonstrates excellent interoccasion and intraobserver reliability and compares favorably with existing RA clinical outcome measures. Synovial volume measurements demonstrated good intraobserver reliability and appeared to be more responsive to synovial change over a 48-hour period in this group of patients. PMID- 12632413 TI - An index of the three core data set patient questionnaire measures distinguishes efficacy of active treatment from that of placebo as effectively as the American College of Rheumatology 20% response criteria (ACR20) or the Disease Activity Score (DAS) in a rheumatoid arthritis clinical trial. AB - OBJECTIVE: To evaluate the capacity of a pooled index of only the 3 patient self report questionnaire measures among the 7 American College of Rheumatology (ACR) core data set (Core Data Set) measures to distinguish efficacy of active treatment of rheumatoid arthritis (RA) with leflunomide or methotrexate versus placebo in a randomized, controlled clinical trial, and to compare the results with those obtained using the ACR 20% response criteria (ACR20), Disease Activity Score (DAS), and other pooled indices. METHODS: The 7 ACR Core Data Set measures of 1) joint swelling, 2) joint tenderness, 3) physician global assessment, 4) erythrocyte sedimentation rate (ESR), 5) functional disability, 6) pain, and 7) patient global assessment were combined into the following 5 pooled indices: "All Core Data Set" (all 7 measures), "Assessor Only" (measures 1-3), "Assessor + ESR" (measures 1-4), "Patient Only" (measures 5-7), and "Patient + ESR" (measures 4 7). The capacity of each of these 5 indices to detect differences between active treatment and placebo treatment was compared with that of the ACR20 and the DAS using 4 different analytic methods, each of which presented advantages and limitations. Agreement of the indices with one another and with the ACR20 and the DAS was analyzed according to pairwise kappa statistics and Z scores in multivariate logistic regression models. RESULTS: Each of the 5 indices, including "Patient Only," had a similar capacity to detect greater efficacy of leflunomide and methotrexate versus placebo in this clinical trial, according to each of 4 methods, at similar levels of statistical and clinical significance. CONCLUSION: A pooled index of patient self-report questionnaire Core Data Set measures appears to be as informative as ACR20 responses, DAS scores, and pooled indices of all and assessor-derived Core Data Set measures for distinguishing between active treatment and placebo treatment in this RA clinical trial. PMID- 12632414 TI - Heterogeneity of response of rheumatoid synovium cell subsets to interleukin-18 in relation to differential interleukin-18 receptor expression. AB - OBJECTIVE: To examine the differential response of rheumatoid arthritis (RA) synovium cell subsets to interleukin-18 (IL-18), the effect of IL-18 on Th1 cytokine production, and the regulation of IL-18 by IL-18 binding protein (IL 18BP). METHODS: RA fibroblast-like synoviocytes were stimulated with IL-1 beta, IL-12, and IL-18, and levels of IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). Expression of IL-18 receptor alpha and beta chains (IL-18R alpha and IL-18R beta, respectively), interferon-gamma (IFN gamma), and IL-17 messenger RNA (mRNA) by peripheral blood mononuclear cells, by total RA synovium cells containing T cells obtained after collagenase digestion, and by RA fibroblast like synoviocytes was determined by reverse transcription-polymerase chain reaction. Levels of IFN gamma were measured by ELISA. RESULTS: IL-1 beta and, less effectively, IL-12 could induce RA fibroblast-like synoviocytes to produce IL-6, but IL-18 failed to have an effect. Although IL-18R alpha mRNA was constitutively expressed by RA fibroblast-like synoviocytes, IL-18R beta could not be detected, either with or without stimulation with IL-1 or IL-12. Total RA synovium cells containing T cells showed a strong expression of both IL-18R alpha and IL-18R beta mRNA, and only IL-18R beta was up-regulated by IL-12. The combination of IL-12 and IL-18 synergistically up-regulated IFN gamma mRNA expression by total RA synovium cells containing T cells, but down-regulated that of IL-17. IL-12-induced IFN gamma production by total RA synovium cells containing T cells was increased by additional IL-18 and decreased by IL-18BP. CONCLUSION: These results indicate that IL-18 plays an important role in RA inflammation and joint destruction via T cells and macrophages, but it does not have a direct effect on fibroblast-like synoviocytes. IL-18BP may be a tool for RA therapy because of its ability to neutralize endogenous IL-18. PMID- 12632415 TI - A poly(ADP-ribose) polymerase haplotype spanning the promoter region confers susceptibility to rheumatoid arthritis. AB - OBJECTIVE: To investigate the association of the poly(ADP-ribose) polymerase 1 (PARP-1) gene promoter polymorphism with rheumatoid arthritis (RA) predisposition. METHODS: An association study with 213 Spanish RA patients and 242 healthy subjects was carried out to investigate the association of all known PARP-1 gene promoter polymorphisms, i.e., a CA microsatellite repeat, a poly(A)(n), and 3 single point mutations (C410T, C1362T, and G1672A), with disease susceptibility. Additionally, we analyzed the distribution of PARP-1 polymorphisms in 58 Spanish families with 1 or more affected members. RESULTS: Upon complete genotyping of the panel of 455 samples, strong linkage disequilibrium was observed among the 5 PARP-1 polymorphisms. Only 2 PARP-1 haplotypes were detected: haplotype A (410T-[A](10)-[CA](10-12)-1362C, which includes short PARP-1 CA alleles) and haplotype B (410C-[A](11)-[CA](13-20) 1362T, always paired with long PARP-1 CA variants). Regarding the G1672A variation, although linkage disequilibrium was detected, it did not seem to be part of the conserved haplotypes described. Haplotype B was statistically overrepresented in the RA patient group compared with the healthy subjects (odds ratio 1.42, 95% confidence interval 1.06-1.91, P = 0.019). In addition, a significant dose effect of PARP-1 haplotype carriage on disease predisposition was observed. Of note, within haplotype B, the PARP-1 CA 97-bp allele was found to be the RA-predisposing marker (odds ratio 2.17, 95% confidence interval 1.27 3.72, P = 0.003, corrected P < 0.05). CONCLUSION: Our results demonstrate the existence of 2 unique PARP-1 haplotypes in the Spanish population and provide the first evidence that PARP-1 haplotypes play a role in susceptibility to RA. PMID- 12632416 TI - Bacterial peptidoglycans but not CpG oligodeoxynucleotides activate synovial fibroblasts by toll-like receptor signaling. AB - OBJECTIVE: To test the hypothesis that bacterial products acting as adjuvants, such as CpG oligodeoxynucleotides (ODNs) and peptidoglycans (PGs), are able to activate synoviocytes, and to determine the involvement of Toll-like receptors (TLRs) in this activation process. METHODS: Cultured synovial fibroblasts obtained from patients with rheumatoid arthritis (RA) or osteoarthritis (OA) were stimulated with CpG ODNs or PGs. The expression of various integrins was determined by fluorescence-activated cell sorting. TLR and matrix metalloproteinase (MMP) messenger RNA (mRNA) was measured by real-time polymerase chain reaction. Additionally, levels of interleukin-6 (IL-6) and IL-8 in the culture supernatants were assessed by enzyme-linked immunosorbent assay. Blocking experiments were performed by adding anti-TLR-2 and anti-TLR-4 monoclonal antibodies to cultures stimulated with bacterial PGs. RESULTS: Incubation of synovial fibroblasts with CpG ODNs resulted in neither up-regulation of the expression of integrins on the cell surface, up-regulation of MMP mRNA expression, nor IL-6 and IL-8 production. However, incubation of RA synovial fibroblasts as well as OA synovial fibroblasts with staphylococcal PGs led to an up-regulation of CD54 (ICAM-1) surface expression and to increased expression of MMP-1, MMP-3, and MMP-13 mRNA. Furthermore, production of the proinflammatory cytokines IL-6 and IL-8 was increased by treatment with PGs. We demonstrated that cultured synovial fibroblasts express low levels of TLR-2 and TLR-9 mRNA. TLR-2 was up-regulated after stimulation with PGs, whereas TLR-9 mRNA remained at baseline levels after stimulation with CpG ODNs. Anti-TLR-2 monoclonal antibodies significantly inhibited production of IL-6 and IL-8 induced by stimulation with PGs. CONCLUSION: We demonstrate that bacterial PGs activate synovial fibroblasts, at least partially via TLR-2, to express integrins, MMPs, and proinflammatory cytokines. Inhibition of TLR signaling pathways might therefore have a beneficial effect on both joint inflammation and joint destruction. PMID- 12632417 TI - Role of the tissue factor pathway in synovial inflammation. AB - OBJECTIVE: Clinical and experimental evidence suggests that extravascular fibrin deposition in arthritic joints is prominent and deleterious. The aim of this study was to investigate the contributions of tissue factor (TF) and its inhibitor, TF pathway inhibitor (TFPI), in arthritis. METHODS: Synovial tissue specimens obtained from 10 patients with rheumatoid arthritis (RA) and 12 patients with osteoarthritis (OA) were scored histologically for inflammation and fibrin content. TF and TFPI levels were assayed at antigenic and functional levels. TF messenger RNA (mRNA) levels were determined using RNase protection assays. The effect of TF inhibition in murine antigen-induced arthritis (AIA) was assessed by administering systemically active site-blocked activated factor VIIa (FVIIai). RESULTS: Functional TF activity was significantly increased in synovial membranes from RA patients compared with those from OA patients. In contrast, no difference in TF mRNA and TF antigenic levels was observed between these 2 groups. This discrepancy can be accounted for by TFPI, because we observed a negative correlation between TF activity and TFPI activity. There was a significant difference between the RA and OA groups in terms of synovial inflammation, with more inflammation observed in the RA group. Most importantly, TF activity was associated with fibrin (P = 0.024) and with histologic inflammation (P = 0.03) scores. In AIA, inhibition of TF-induced coagulation by FVIIai led, on day 9 of arthritis, to decreased synovial thickness and decreased articular cartilage damage, although only the latter difference between controls and treated mice reached significance (P < 0.04). Finally, in FVIIai-treated mice, there was a strong negative association between the prothrombin time and intraarticular fibrin deposition. CONCLUSION: Our results show that TF expression in arthritic synovial tissue favors extravascular coagulation and may play a role in inflammation in RA. In this context, TF inhibitors may be of therapeutic value. PMID- 12632418 TI - Expression of pleiotrophin, an embryonic growth and differentiation factor, in rheumatoid arthritis. AB - OBJECTIVE: Pleiotrophin (PTN), a 15.3-kd heparin-binding peptide, is expressed in mesodermal and neuroectodermal cells during development, but rarely in adult tissues. Since developmentally regulated factors often reappear during disease, we sought to determine whether there was PTN expression in the synovial membranes of patients with rheumatoid arthritis (RA). METHODS: PTN messenger RNA expression was assayed by quantitative reverse transcriptase-polymerase chain reaction. The protein was localized by immunohistochemistry and quantified by enzyme-linked immunosorbent assay (ELISA). Effects of PTN on cell proliferation in vitro were determined by DNA measurements. RESULTS: PTN expression in normal adult synovial membranes and cartilage was barely detectable. However, PTN was strongly up regulated in synovial tissues from patients with RA. In contrast, samples from patients with pyogenic arthritis had moderate PTN levels, and those from patients with osteoarthritis had only a slight increase in PTN, as measured by ELISA. In RA patients, PTN was localized primarily in synoviocytes but was also found in endothelial cells of blood vessels. In cultured mouse fibroblasts used as a model, PTN expression was up-regulated by tumor necrosis factor alpha and was more weakly up-regulated by epidermal growth factor. Recombinant PTN stimulated the proliferation of cultured human synoviocytes and the monocyte cell line THP 1, but not human dermal fibroblasts, in which PTN increased the synthesis of vascular endothelial growth factor. CONCLUSION: In addition to certain types of cancer, the embryonic growth and differentiation factor PTN is expressed in adults with inflammatory diseases, in particular, RA. Proinflammatory cytokines enhance the expression of PTN. Thus, we propose that PTN is a further paracrine angiogenesis and growth factor for synovial cells in RA. PMID- 12632419 TI - Specific increase in enzymatic activity of adenosine deaminase 1 in rheumatoid synovial fibroblasts. AB - OBJECTIVE: Adenosine deaminase (ADA; EC 3.5.4.4) activity is elevated in the synovial fluid (SF) of patients with rheumatoid arthritis (RA). Since the antiinflammatory effect of methotrexate is reportedly associated with increased levels of extracellular adenosine, the present study was undertaken to clarify the role of 2 ADA isozymes, ADA1 and ADA2, in the pathogenesis of RA. METHODS: The activities of ADA1 and ADA2 were measured in SF from RA and osteoarthritis (OA) patients, in sera from RA patients, and in lysates prepared from mononuclear and polymorphonuclear cells from SF from RA patients, peripheral blood from RA patients, and fibroblast-like synoviocytes (FLS) from RA and OA patients. Also measured were the effects of proinflammatory cytokines on ADA1 activity and ADA messenger RNA (mRNA) expression in RA FLS, as determined using real-time polymerase chain reaction. The adenosine concentration in RA SF was measured by radioimmunoassay. RESULTS: The adenosine concentration in RA SF ranged from 0.027 microM to 0.508 microM (mean +/- SD 0.156 +/- 0.132 microM). At those concentrations, ADA1 would be expected to be functionally dominant due to its higher affinity for adenosine. ADA1 activity in RA SF (mean +/- SD 14.4 +/- 8.5 IU/liter) was significantly higher than that in OA SF (3.0 +/- 1.1 IU/liter) or RA sera (3.0 +/- 0.6 IU/liter); moreover, ADA1 activity in RA FLS lysate was the highest among the cell lysates tested. Proinflammatory cytokines did not affect ADA1 activity or ADA mRNA expression in RA FLS. CONCLUSION: Elevated ADA1 activity is an intrinsic characteristic of RA FLS, which likely contributes to the pathogenesis of RA by neutralizing the antirheumatic properties of endogenous adenosine. PMID- 12632420 TI - Ethnic and sex differences in serum levels of cartilage oligomeric matrix protein: the Johnston County Osteoarthritis Project. AB - OBJECTIVE: Previous descriptions of potential biomarkers of osteoarthritis (OA) have been limited to Caucasians. In the present study, we examined associations between serum levels of cartilage oligomeric matrix protein (COMP) and ethnicity (African American or Caucasian) and sex in the Johnston County Osteoarthritis Project, a population-based study of OA in rural North Carolina. METHODS: All African Americans and a randomly selected sample of Caucasians who had available sera and either no radiographic evidence of knee or hip OA according to the Kellgren/Lawrence (K/L) system (K/L grade 0) or radiographic evidence of knee OA (K/L grade 2 or higher) were included. Serum COMP levels were quantified by sandwich enzyme-linked immunosorbent assay, using monoclonal antibodies 16-F12 and 17-C10. Linear regression models were used to assess relationships between serum levels of natural log-transformed COMP (ln COMP) and ethnicity and sex, controlling for age, height, body mass index (BMI), radiographic OA, and the presence of other symptomatic joints. Radiographic OA was defined in separate models as the presence, severity, and laterality of radiographic knee OA, the co occurrence of radiographic knee and hip OA, and the number of knees and hips with radiographic OA. RESULTS: The 769 subjects in the study sample had a mean +/- SD age of 62 +/- 10.3 years. Levels of ln COMP were associated with age, BMI, and all definitions of radiographic OA (P = 0.0001), and varied by ethnicity and sex. In adjusted models, ln COMP was higher in African American women than in Caucasian women (P = 0.003) and higher in Caucasian men than Caucasian women (P = 0.0001). There were no statistically significant differences in serum ln COMP levels between African American men and women. CONCLUSION: Serum COMP levels vary by ethnicity and sex. These factors should be considered in the derivation of standards using this, and possibly other, potential biomarkers of OA. PMID- 12632421 TI - Comparison of tibial cartilage volume and radiologic grade of the tibiofemoral joint. AB - OBJECTIVE: To compare tibial cartilage volume as measured by magnetic resonance imaging (MRI) with radiologic assessment of the tibiofemoral joint. METHODS: The MRI-determined tibial cartilage volume was compared with the radiologic grade of individual features of osteoarthritis (osteophytes and joint space narrowing [JSN]) in 252 subjects (mean +/- SD age 60.2 +/- 10 years, 62% female) who were participating in studies of knee cartilage. RESULTS: JSN seen on both medial and lateral radiographs of the tibiofemoral joint was inversely associated with the respective tibial cartilage volume. This inverse relationship was strengthened with adjustment for age, sex, body mass index (BMI), and bone size. After adjustment for these confounders, for every increase in JSN grade (0-3), the medial tibial cartilage volume was reduced by 257 mm(3) (95% confidence interval [95% CI] 193-321) and the lateral tibial cartilage volume by 396 mm(3) (95% CI 283-509). The relationship between mean cartilage volume and radiologic grade of JSN was linear. Based on results in the subgroup of subjects with normal radiographic findings, we have proposed a model to estimate average "normal" cartilage volume in men and women for a given age, BMI, and bone size. CONCLUSION: The results of this study demonstrate a strong negative, linear association between medial and lateral tibial cartilage volume and increasing grade of JSN. Using data from radiographically normal subjects, we have proposed a simple model for estimating "normal" cartilage volume. However, larger studies will be needed to confirm these findings and to determine whether they are valid in younger subjects. PMID- 12632422 TI - Microenvironment regulation of extracellular signal-regulated kinase activity in chondrocytes: effects of culture configuration, interleukin-1, and compressive stress. AB - OBJECTIVE: To compare extracellular signal-regulated kinase (ERK) activity in response to interleukin-1 (IL-1) in chondrocytes under various culture configurations designed for the study of cartilage biology and repair, and also in response to dynamic load for chondrocytes in cartilage. METHODS: Isolated bovine articular chondrocytes were maintained in serum-supplemented medium under 4 culture configurations: high-density monolayer, attached to a cut surface of cartilage, within tissue-engineered constructs, or within intact cartilage explants. Samples were subjected to a change of medium with or without IL-1. Cartilage explants were also subjected to dynamic compression. RESULTS: In chondrocyte monolayers, both basal and IL-1-stimulated ERK activities were similarly elevated at 0.5 hours after medium change, diminishing by 74% after 16 hours. In contrast, chondrocytes in other culture configurations exhibited lower basal levels of ERK activity and a moderate activation of ERK in response to IL-1 that was sustained over the 16-hour treatment time. The dynamic component of loading of cartilage explants led to a 5-fold activation of ERK, compared with free-swelling controls, that was indistinguishable from the effects of IL-1. CONCLUSION: ERK signaling in response to IL-1 in chondrocyte monolayers exhibited a pattern that was distinct from that in other culture systems, suggesting that the extracellular matrix plays an important regulatory role in modulating the response to extracellular stimuli. Since IL-1 and dynamic loading have distinct effects on chondrocyte biosynthesis, signaling pathways other than ERK participate in the chondrocyte responses to these stimuli. PMID- 12632423 TI - Mitochondrial respiratory activity is altered in osteoarthritic human articular chondrocytes. AB - OBJECTIVE: Osteoarthritis (OA) is a degenerative rheumatic disease that is associated with extracellular matrix degradation and chondrocyte apoptosis in the articular cartilage. The role of mitochondria in degenerative diseases is widely recognized. We undertook this study to evaluate mitochondrial function in normal and OA chondrocytes and to examine age-related changes in mitochondria. METHODS: Mitochondrial function was evaluated by analyzing respiratory chain enzyme complexes and citrate synthase (CS) activities as well as changes in mitochondrial membrane potential (Delta Psi m). The activities of mitochondrial respiratory chain complexes (complex I: rotenone-sensitive NADH-coenzyme Q(1) reductase; complex II: succinate dehydrogenase; complex III: antimycin-sensitive ubiquinol cytochrome c reductase; and complex IV: cytochrome c oxidase) and CS were measured in human articular chondrocytes isolated from OA and normal cartilage. Delta Psi m was measured by JC-1 using flow cytometry. Statistical analysis was performed using the Mann-Whitney U test and Student's t-test as well as several models of multiple linear regression. RESULTS: OA articular chondrocytes had reduced activities of complexes II and III compared with cells from normal cartilage. However, the mitochondrial mass was increased in OA. Cultures of OA chondrocytes contained a higher proportion of cells with de energized mitochondria. We found no relationship between mitochondrial function and donor age either in normal or in OA chondrocytes. CONCLUSION: These findings suggest the involvement of mitochondrial function in the pathophysiology of OA. Cartilage degradation by OA and cartilage aging may be two different processes. PMID- 12632424 TI - Inhibition of cartilage degradation: a combined tissue engineering and gene therapy approach. AB - OBJECTIVE: To determine if tissue-engineered cartilage can be protected from cytokine-induced degradation using a gene therapy approach. METHODS: Chemical and pantropic retroviral gene transfer methodologies were compared for their ability to introduce a luciferase reporter gene into adult bovine cartilage chondrocytes grown in monolayer. Pantropic retrovirus was then used to transduce these cells with human tissue inhibitor of metalloproteinases 1 (TIMP-1), and the stability of expression in monolayer or pellet culture was monitored for 6 weeks. Untransduced and TIMP-1-transduced cells were also used to tissue engineer 3 dimensional cartilage constructs that were then challenged with interleukin-1 (IL 1) for 4 weeks. Conditioned media and residual cartilage were collected for analysis of matrix components, including type II collagen and proteoglycans, and for TIMP-1 production and matrix metalloproteinase (MMP) activity. RESULTS: Chemical transfection of adult bovine chondrocytes gave rise to short-lived reporter expression that was virtually undetectable after 4 weeks of culture. In contrast, pantropic retroviral transduction gave rise to stable expression that persisted at a high level for at least 6 weeks. Pantropic transduction of the cells with TIMP-1 gave rise to similar long-term expression, both in monolayer and pellet cultures. TIMP-1-transduced tissue-engineered cartilage also retained TIMP-1 expression for an additional 4 weeks of culture in the presence of IL-1. Compared with control samples, TIMP-1-transgenic cartilage resisted the catabolic effects of IL-1, with MMP activity reduced to basal levels and a decreased loss of type II collagen. CONCLUSION: Pantropic retroviral transduction permits long term expression of potentially therapeutic transgenes in adult tissue-engineered cartilage. While TIMP-1 transduction could be used to prevent collagen breakdown, alternative transgenes may be necessary to protect cartilage proteoglycans. PMID- 12632425 TI - A short course of BG9588 (anti-CD40 ligand antibody) improves serologic activity and decreases hematuria in patients with proliferative lupus glomerulonephritis. AB - OBJECTIVE: CD40-CD40 ligand (CD40L) interactions play a significant role in the production of autoantibodies and tissue injury in lupus nephritis. We performed an open-label, multiple-dose study to evaluate the safety, efficacy, and pharmacokinetics of BG9588, a humanized anti-CD40L antibody, in patients with proliferative lupus nephritis. The primary outcome measure was 50% reduction in proteinuria without worsening of renal function. METHODS: Twenty-eight patients with active proliferative lupus nephritis were scheduled to receive 20 mg/kg of BG9588 at biweekly intervals for the first 3 doses and at monthly intervals for 4 additional doses. Safety evaluations were performed on all patients. Eighteen patients receiving at least 3 doses were evaluated for efficacy. RESULTS: The study was terminated prematurely because of thromboembolic events occurring in patients in this and other BG9588 protocols (2 myocardial infarctions in this study). Of the 18 patients for whom efficacy could be evaluated, 2 had a 50% reduction in proteinuria without worsening of renal function. Mean reductions of 38.9% (P < 0.005), 50.1% (P < 0.005), and 25.3% (P < 0.05) in anti-double stranded DNA (anti-dsDNA) antibody titers were observed at 1, 2, and 3 months, respectively, after the last treatment. There was a significant increase in serum C3 concentrations at 1 month after the last dose (P < 0.005), and hematuria disappeared in all 5 patients with significant hematuria at baseline. There were no statistically significant reductions in lymphocyte count or serum immunoglobulin, anticardiolipin antibody, or rubella IgG antibody concentrations after therapy. CONCLUSION: A short course of BG9588 treatment in patients with proliferative lupus nephritis reduces anti-dsDNA antibodies, increases C3 concentrations, and decreases hematuria, suggesting that the drug has immunomodulatory action. Additional studies will be needed to evaluate its long term effects. PMID- 12632426 TI - Randomized study of subcutaneous low molecular weight heparin plus aspirin versus intravenous immunoglobulin in the treatment of recurrent fetal loss associated with antiphospholipid antibodies. AB - OBJECTIVE: To compare the 2 most efficacious therapeutic regimens, intravenous immunoglobulin (IVIG) and anticoagulation with low molecular weight (LMW) heparin plus low-dose aspirin, in women with recurrent pregnancy loss associated with antiphospholipid antibodies (aPL). METHODS: We examined 40 women with recurrent abortion (at least 3 occurrences) and repeatedly positive test results for anticardiolipin or lupus anticoagulant. The subjects were randomly assigned to treatment with IVIG or LMW heparin plus low-dose aspirin. Both therapies were started when the women were pregnant as documented by a positive urine test. IVIG was stopped at the thirty-first week of gestation, aspirin at the thirty-fourth week, and heparin at the thirty-seventh week. The primary outcome of interest was the rate of live births with the 2 treatments. RESULTS: The characteristics of the 2 groups were similar at the time of randomization. The women treated with LMW heparin plus low-dose aspirin had a higher rate of live births (84%) than those treated with IVIG (57%). CONCLUSION: Treatment with LMW heparin plus low dose aspirin should be considered as the standard therapy for recurrent pregnancy loss due to aPL. PMID- 12632427 TI - Asymptomatic avascular necrosis in patients with primary antiphospholipid syndrome in the absence of corticosteroid use: a prospective study by magnetic resonance imaging. AB - OBJECTIVE: To evaluate the prevalence of avascular necrosis (AVN), using magnetic resonance imaging (MRI), in patients with primary antiphospholipid syndrome (APS) and in patients with systemic lupus erythematosus (SLE), with or without anticardiolipin antibodies (aCL), who are asymptomatic for AVN and have not taken corticosteroids. METHODS: Seventy-nine subjects who were asymptomatic for AVN were evaluated by MRI of the femoral heads: 30 patients with primary APS who had never received corticosteroids, 19 SLE patients who had never received steroids (divided into 2 groups, aCL positive and aCL negative, in order to examine any association between AVN and aCL), and 30 healthy subjects who were age- and sex matched with patients with primary APS. Established MRI criteria were used for a diagnosis of AVN. RESULTS: Asymptomatic AVN was evident in 6 (20%) of 30 patients with primary APS: 3 of them (1 man, 2 women) had intermediate bilateral AVN, and 3 (all women) had early AVN (bilateral in 1 patient). Results of hip and pelvis radiography and dynamic scintigraphy were negative. Followup MRI 6 months later revealed no changes. At the time of the initial MRI examination, the mean (+/-SD) age of patients in whom AVN was identified was 31.2 +/- 7.3 years, and that of patients without AVN was 42.4 +/- 11.9 years (P = 0.036). Livedo reticularis occurred significantly more commonly in the group with AVN (P = 0.041). None of the healthy subjects and none of the patients with SLE demonstrated AVN on MRI. CONCLUSION: AVN can be detected by MRI in 20% of patients with primary APS. Younger patients tend to develop AVN more frequently than do older patients, and the presence of livedo reticularis may identify individuals at risk for AVN. Clinicians should be aware of this possible clinical manifestation of primary APS, because early diagnosis can lead to early intervention. PMID- 12632428 TI - Identification and characterization of a peptide mimetic that may detect a species of disease-associated anticardiolipin antibodies in patients with the antiphospholipid syndrome. AB - OBJECTIVE: To test the feasibility of applying a mimetic (specific for a patient derived prothrombotic anticardiolipin antibody [aCL]) to study the homologous, disease-associated aCL in patients with antiphospholipid syndrome (APS). METHODS: We used the CL15 monoclonal aCL to screen 17 phage-display peptide libraries. Peptides (corresponding to recurrent peptide sequences) and their derivatives were synthesized and analyzed for binding to CL15 and for their abilities to inhibit CL15 from binding to cardiolipin. A peptide was chosen and used to study CL15-like IgG aCL in plasma samples from patients with APS, patients with systemic lupus erythematosus (SLE) but without APS, and normal healthy donors. RESULTS: Library screening with CL15 yielded 4 recurrent peptide sequences. Analyses of peptides showed that peptide CL154C reacted with antibody CL15 and inhibited binding of CL15 to cardiolipin, indicating that peptide CL154C may be a peptide mimetic for the CL15 aCL. Initial studies with plasma samples revealed that CL154C-reactive IgG was present (positivity defined as the mean + 3 SD optical density of the 25 normal controls) in 15 of 21 APS patients and 1 of 12 SLE patients. CONCLUSION: These findings suggest that it is feasible to develop a specific enzyme-linked immunosorbent assay for each immunologically and functionally distinct disease-associated aCL. Additional testing of CL154C with a larger number of APS patients and SLE patients, as well as identification of peptide mimetics for each distinct aCL, will reveal the diagnostic potential of CL154C and other mimetics in identifying patients with aCL who are at risk of developing life-threatening thrombosis. PMID- 12632429 TI - Hydralazine may induce autoimmunity by inhibiting extracellular signal-regulated kinase pathway signaling. AB - OBJECTIVE: To determine whether hydralazine might decrease DNA methyltransferase (DNMT) expression and induce autoimmunity by inhibiting extracellular signal regulated kinase (ERK) pathway signaling. METHODS: The effect of hydralazine on DNMT was tested in vitro using enzyme inhibition studies, and in vivo by measuring messenger RNA (mRNA) levels and enzyme activity. Effects on ERK, c-Jun N-terminal kinase, and p38 pathway signaling were tested using immunoblotting. Murine T cells treated with hydralazine or an ERK pathway inhibitor were injected into mice and anti-DNA antibodies were measured by enzyme-linked immunosorbent assay. RESULTS: In vitro, hydralazine did not inhibit DNMT activity. Instead, hydralazine inhibited ERK pathway signaling, thereby decreasing DNMT1 and DNMT3a mRNA expression and DNMT enzyme activity similar to mitogen-activated protein kinase kinase (MEK) inhibitors. Inhibiting T cell ERK pathway signaling with an MEK inhibitor was sufficient to induce anti-double-stranded DNA antibodies in a murine model of drug-induced lupus, similar to the effect of hydralazine. CONCLUSION: Hydralazine reproduces the lupus ERK pathway signaling abnormality and its effects on DNMT expression, and inhibiting this pathway induces autoimmunity. Hydralazine-induced lupus could be caused in part by inducing the same ERK pathway signaling defect that occurs in idiopathic lupus. PMID- 12632430 TI - Attenuation of autoimmune disease in Fas-deficient mice by treatment with a cytotoxic benzodiazepine. AB - OBJECTIVE: Elimination of autoreactive cells relies on Fas-dependent activation induced cell death mechanisms, an important component of peripheral tolerance. Defects in Fas or its cognate ligand lead to inefficient activation-induced cell death and are specific causes of lymphoproliferative and autoimmune diseases. The present study was undertaken to investigate a novel 1,4-benzodiazepine (Bz-423) that induces apoptosis and limits autoimmune disease in NZB/NZW mice, to determine its activity against lupus-like disease associated with defective Fas expression. We investigated the Fas-dependence of its cytotoxic actions, its therapeutic potential in mice deficient in Fas, and its therapeutic mechanism of action. METHODS: Primary lymphocytes isolated from Fas-deficient MRL/MpJ-Fas(lpr) (MRL-lpr) mice were tested for sensitivity to Bz-423. Bz-423 was administered to MRL-lpr mice for short (1-week) or long (14-week) periods, and its effects on cell survival were determined along with measures of nephritis, arthritis, antibody titers, and Th subpopulations. BALB/c mice were similarly treated to determine if Bz-423 alters normal immune functions in vivo. RESULTS: Administration of Bz-423 to MRL-lpr mice significantly reduced autoimmune disease including glomerulonephritis and arthritis. Treatment was associated with decreases in CD4+ T cells and an alteration in the Th1/Th2 balance. At the therapeutic dosage, Bz-423 did not interfere with normal T and B cell responses in BALB/c mice, suggesting that this agent is not globally immunosuppressive. CONCLUSION: Bz-423 is a novel immunomodulatory agent that is active against disease even in the context of defective Fas signaling. It is a leading compound for further investigation into the development of selective therapies for lupus. PMID- 12632431 TI - Outcome in adults with juvenile idiopathic arthritis: a quality of life study. AB - OBJECTIVE: To evaluate quality of life (QOL) in adults with juvenile idiopathic arthritis (JIA), using validated measures of functional disability and generic health status, and to quantify their educational attainment and employment status. METHODS: The adult rheumatology departmental database was used to identify patients. Functional disability and generic health status/QOL were assessed by the Health Assessment Questionnaire (HAQ) and the Short Form 36-item health profile (SF-36), respectively. Educational achievement and employment status were assessed by questionnaire. RESULTS: Complete data were available for 82 of the 101 patients identified. The median age of patients was 30 years, and the median disease duration was 21 years. No deaths were recorded. All subtypes of JIA were represented. Thirty-nine percent of patients had active disease (based on the physician global assessment scale score). The median HAQ score was 1.125 (range 0-3). SF-36 scores for bodily pain, general health, physical functioning, vitality, emotion, and social isolation were significantly worse in patients compared with controls, and this trend increased with increasing age of the patients and disease duration. The SF-36 mental summation scores of patients were low compared with those of controls, for all subtypes of JIA, and this finding was independent of the degree of functional disability (by HAQ and SF-36 physical summation scores). The educational attainment of patients was comparable to that of local controls, but unemployment rates for patients were 3-fold higher than those for controls. CONCLUSION: This is the largest study in which the SF-36 was used to assess generic health status and QOL in adults with JIA. Many patients had active disease in adulthood, and although the physical outcome of adults with JIA is relatively good, a profound effect on generic health status and QOL was demonstrated for all types of JIA. Furthermore, despite excellent educational attainment, there was a high rate of unemployment among patients. PMID- 12632432 TI - A rare case of juxtaarticular osteoid osteoma of the calcaneus initially misdiagnosed as juvenile chronic arthritis. AB - Juxtaarticular osteoid osteoma is frequently misdiagnosed because the symptoms may mimic arthritis, and radiographs may not be characteristic. A rare case of subtalar pain lasting 5 years in a female teenager is presented here. The initial diagnosis was monarticular juvenile chronic arthritis. Family history was misleading because her mother had rheumatoid arthritis (RA). PMID- 12632433 TI - Down-regulation of the nonspecific and antigen-specific T cell cytokine response in ankylosing spondylitis during treatment with infliximab. AB - OBJECTIVE: Treatment of active ankylosing spondylitis (AS) with the monoclonal tumor necrosis factor alpha (TNF alpha) antibody infliximab is highly clinically effective. This study was undertaken to investigate the precise mechanism of action of anti-TNF alpha treatment in AS. METHODS: Cytokine expression of CD4+ and CD8+ T cells was investigated before and 6 and 12 weeks after the start of treatment in 10 patients treated with infliximab, and before and after 6 weeks of treatment and 6 weeks after placebo was switched to infliximab in 10 patients treated initially with placebo. Peripheral blood mononuclear cells (PBMCs) were stimulated for 6 hours either nonspecifically with phorbol myristate acetate (PMA)/ionomycin or antigen specifically with a pool of 46 overlapping 18-mer peptides derived from the G1 domain of aggrecan. Cells were stained for T cell surface markers CD4 and CD8 and for the intracellular cytokines interferon-gamma (IFN gamma), TNF alpha, interleukin-4 (IL-4), and IL-10. Positive cells were quantified by flow cytometry. For monocyte-derived cytokines, PBMCs were stimulated with lipopolysaccharide (LPS) for 18 hours and TNF alpha and IL-10 in the supernatant were measured by enzyme-linked immunosorbent assay. RESULTS: Compared with baseline, infliximab treatment induced a significant decrease at 12 weeks in the number of CD4+ and CD8+ T cells that were positive for IFN gamma and TNF alpha upon PMA/ionomycin stimulation (P = 0.005). A significant reduction had already begun to occur at 6 weeks. No change in the percent IFN gamma or TNF alpha positivity among CD4+ and CD8+ subpopulations was observed after 6 weeks in patients treated with placebo. However, when these patients began infliximab treatment after 6 weeks of receiving placebo, there was a similar significant decrease in IFN gamma and TNF alpha production by CD4+ and CD8+ T cells (P < 0.05). Furthermore, infliximab treatment induced a significant reduction in the number of IFN gamma+ and TNF alpha+ CD8+ T cells (P = 0.005 at week 6 and week 12) after antigen-specific in vitro stimulation with G1-derived peptides. Between group analysis showed that the change in the expression of IFN gamma and TNF alpha in both CD4+ and CD8+ T cells was significantly different between the infliximab and placebo groups (P = 0.001 for all variables). There was no change in the number of IL-10+ or IL-4+ T cells during treatment. No significant change in the production of TNFalpha and IL-10 upon in vitro stimulation of PBMCs with LPS was detectable during infliximab treatment. CONCLUSION: Infliximab down regulates both IFN gamma and TNF alpha secreted by T cells but does not induce a change in cytokines produced by monocytes during 3 months of treatment. This is likely to be a relevant mechanism for the clinical efficacy of this therapy. PMID- 12632434 TI - Novel genetic markers in the 5'-flanking region of ANKH are associated with ankylosing spondylitis. AB - OBJECTIVE: To use a candidate gene approach for the identification of genetic markers that are significantly linked to and associated with ankylosing spondylitis (AS). METHODS: We searched for novel polymorphisms in the ANKH gene (human homolog of the murine progressive ankylosis gene) and genotyped 2 polymorphic sites, one in the 5'-noncoding region and the other in the promoter region of ANKH, using DNA from affected (n = 273) and unaffected (n = 112) individuals from 124 AS families. We used these ANKH and other nearby polymorphisms to perform linkage and family-based association analyses. RESULTS: We identified 2 novel polymorphic sites: one in the 5'-noncoding region of ANKH involving 1-2 copies of an 8-bp repeat (denoted as ANKH-OR), and the other in the promoter region involving different copy numbers of a triplet repeat (denoted as ANKH-TR). ANKH-OR and ANKH-TR were in complete linkage disequilibrium. Five markers (D5S1953, ANKH-TR, ANKH-OR, D5S1954, and D5S1963) were used for both the linkage and association analyses. Multipoint linkage analysis of 124 AS families showed a modest level of significance (nonparametric linkage score 2.15; P = 0.015) at the ANKH region. The contribution of ANKH to AS susceptibility (lambda(s)) was 1.9. A family-based association study on the same AS families revealed that both ANKH-OR allele 1 and ANKH-TR allele 7 were significantly associated with disease, assuming an additive model (for ANKH-OR allele 1, P = 0.03; for ANKH-TR allele 7, P = 0.04). CONCLUSION: Our results indicate that ANKH OR and ANKH-TR are novel genetic markers that are significantly associated with AS. PMID- 12632435 TI - Selective stimulation of collagen synthesis in the presence of costimulatory insulin signaling by connective tissue growth factor in scleroderma fibroblasts. AB - OBJECTIVE: To examine the mechanism of collagen induction by connective tissue growth factor (CTGF), a profibrotic cytokine overexpressed in the skin of patients with systemic sclerosis (SSc). METHODS: Dermal fibroblasts from 7 SSc patients and 7 matched healthy adult donors were stimulated with CTGF in the presence or absence of the culture-medium supplement, insulin-transferrin selenium (ITS). Expression of collagen protein was analyzed by a (3)H-proline incorporation assay. To identify the signaling pathways mediating CTGF induction of collagen, pharmacologic inhibitors were used, including rottlerin, a protein kinase C delta (PKC delta) inhibitor. RESULTS: Collagen levels in both SSc and normal fibroblasts were increased after treatment with transforming growth factor beta in serum-free medium, whereas no stimulation was observed following addition of CTGF. In the presence of ITS, CTGF (2.5 ng/ml) potently stimulated collagenous protein levels in SSc cell lines (n = 5); however, CTGF was not stimulatory in the majority of normal fibroblasts (n = 6). ITS alone induced collagen levels in normal fibroblasts to the levels observed in SSc skin fibroblasts, thereby diminishing the hallmark difference in basal collagen levels in these cell types. Insulin was the ITS component responsible for promoting the basal and CTGF stimulation of collagenous proteins. Rottlerin, the PKC delta inhibitor, down regulated collagen synthesis in normal and SSc fibroblasts cultured in ITS, and inhibited the stimulatory effects of CTGF in cooperation with insulin or of insulin (500 ng/ml) alone. CONCLUSION: Increased responsiveness of SSc fibroblasts to CTGF-mediated collagen synthesis requires the costimulatory activation of insulin signaling pathways to induce matrix production. Blockade of this effect via rottlerin may suggest that PKC delta is a downstream signaling molecule necessary for CTGF stimulation of collagen synthesis. PMID- 12632436 TI - Mapping the HLA association in Behcet's disease: a role for tumor necrosis factor polymorphisms? AB - OBJECTIVE: Experimental evidence suggests that inappropriate regulation of tumor necrosis factor alpha (TNF alpha) may play a role in the pathogenesis of Behcet's disease (BD). This is supported by recent reports highlighting the efficacy of anti-TNF alpha agents in the treatment of this disease. The TNF gene is encoded in the class III region of the HLA complex adjacent to HLA-B. This genetic proximity to a gene that is already widely implicated in disease susceptibility led us to investigate the association between TNF promoter polymorphisms and susceptibility to BD. METHODS: We studied 133 UK white Caucasoid patients with BD and 354 healthy controls. We attempted to dissect the contribution of individual polymorphisms in this gene-dense region by linkage disequilibrium mapping across 6 adjacent genes. RESULTS: We report a novel association with the TNF promoter allele TNF-1031C. Subsequent analysis identified 2 extended HLA haplotypes associated with BD. One of them contained the previously recognized susceptibility gene HLA-B*51, while the other was defined by HLA-B*5701. Both of these haplotypes contained the TNF promoter polymorphism -1031C, an allele that was associated with disease even in individuals who did not carry either HLA-B*51 or HLA-B*5701. CONCLUSION: The TNF-1031C allele is independently associated with susceptibility to BD in Caucasoid patients. Further studies will be required to determine the functional effects of this polymorphism, its influence in disease pathogenesis, and its role in other ethnic groups. PMID- 12632437 TI - Are environmental factors important in primary systemic vasculitis? A case control study. AB - OBJECTIVE: To investigate the association between primary systemic vasculitis (PSV) and environmental risk factors. METHODS: Seventy-five PSV cases and 273 controls (220 nonvasculitis, 19 secondary vasculitis, and 34 asthma controls) were interviewed using a structured questionnaire. Factors investigated were social class, occupational and residential history, smoking, pets, allergies, vaccinations, medications, hepatitis, tuberculosis, and farm exposure in the year before symptom onset (index year). The Standard Occupational Classification 2000 and job-exposure matrices were used to assess occupational silica, solvent, and metal exposure. Stepwise multiple logistic regression was used to calculate the odds ratio (OR) and 95% confidence interval (95% CI) adjusted for potential confounders. Total PSV, subgroups (47 Wegener's granulomatosis [WG], 12 microscopic polyangiitis, 16 Churg-Strauss syndrome [CSS]), and antineutrophil cytoplasmic antibody (ANCA)-positive cases were compared with control groups. RESULTS: Farming in the index year was significantly associated with PSV (OR 2.3 [95% CI 1.2-4.6]), with WG (2.7 [1.2-5.8]), with MPA (6.3 [1.9-21.6]), and with perinuclear ANCA (pANCA) (4.3 [1.5-12.7]). Farming during working lifetime was associated with PSV (2.2 [1.2-3.8]) and with WG (2.7 [1.3-5.7]). Significant associations were found for high occupational silica exposure in the index year (with PSV 3.0 [1.0-8.4], with CSS 5.6 [1.3-23.5], and with ANCA 4.9 [1.3-18.6]), high occupational solvent exposure in the index year (with PSV 3.4 [0.9-12.5], with WG 4.8 [1.2-19.8], and with classic ANCA [cANCA] 3.9 [1.6-9.5]), high occupational solvent exposure during working lifetime (with PSV 2.7 [1.1-6.6], with WG 3.4 [1.3-8.9], and with cANCA 3.3 [1.0-10.8]), drug allergy (with PSV 3.6 [1.8-7.0], with WG 4.0 [1.8-8.7], and with cANCA 4.7 [1.9-11.7]), and allergy overall (with PSV 2.2 [1.2-3.9], with WG 2.7 [1.4-5.7]). No other significant associations were found. CONCLUSION: A significant association between farming and PSV has been identified for the first time. Results also support previously reported associations with silica, solvents, and allergy. PMID- 12632438 TI - Serum osteoprotegerin and its ligand in Paget's disease of bone: relationship to disease activity and effect of treatment with bisphosphonates. AB - OBJECTIVE: To test the following hypotheses: 1) osteoprotegerin (OPG) and receptor activator of nuclear factor kappa B ligand (RANKL) serum levels in patients with Paget's disease are related to disease activity and are different from those in healthy individuals; 2) interleukin-6 (IL-6), a cytokine that has been shown to have higher levels in Paget's disease, modulates these factors; and 3) the antiresorptive effect of bisphosphonates in Paget's disease of bone may be mediated through these local factors. METHODS: The study group comprised 31 patients with Paget's disease who received 400 mg/day of oral tiludronate for 3 months. Serum levels of OPG, RANKL, IL-6, bone alkaline phosphatase (AP), N terminal type I procollagen propeptide, urinary N-terminal crosslinking telopeptide of type I collagen, and urinary alpha-C-terminal crosslinking telopeptide of type I collagen were measured at baseline and 1 month after the end of therapy. In addition, the RANKL:OPG ratio was calculated, and disease activity was evaluated at baseline by quantitative bone scintigraphy. RESULTS: Mean baseline OPG values were higher in patients with Paget's disease than in healthy control subjects (P < 0.005), but RANKL and IL-6 values and RANKL:OPG ratios in the 2 groups were similar. OPG concentrations decreased significantly after treatment with tiludronate (P < 0.005), whereas no significant changes were observed in serum RANKL values. No correlation was found between either bone markers or quantitative scintigraphic indices and serum levels of OPG, RANKL, IL 6, and RANKL:OPG ratios. Serum OPG decreased significantly only in those patients with baseline OPG values >4.1 pM/liter. CONCLUSION: Serum OPG increases in Paget's disease and decreases after treatment with tiludronate, especially in patients with the highest OPG values. In contrast, RANKL serum levels and RANKL:OPG ratios are unmodified in patients with Paget's disease. Although serum OPG, RANKL, and IL-6 values were unrelated to disease activity, the increase in OPG may reflect a protective mechanism of the skeleton to compensate for increased bone resorption. PMID- 12632439 TI - Intraarticular corticosteroids, supervised physiotherapy, or a combination of the two in the treatment of adhesive capsulitis of the shoulder: a placebo-controlled trial. AB - OBJECTIVE: To compare the efficacy of a single intraarticular corticosteroid injection, a supervised physiotherapy program, a combination of the two, and placebo in the treatment of adhesive capsulitis of the shoulder. METHODS: Ninety three subjects with adhesive capsulitis of <1 year's duration were randomized to 1 of 4 treatment groups: group 1, corticosteroid injection (triamcinolone hexacetonide 40 mg) performed under fluoroscopic guidance followed by 12 sessions of supervised physiotherapy; group 2, corticosteroid injection alone; group 3, saline injection followed by supervised physiotherapy; or group 4, saline injection alone (placebo group). All subjects were taught a simple home exercise program. Subjects were reassessed after 6 weeks, 3 months, 6 months, and 1 year. The primary outcome measure was improvement in the Shoulder Pain and Disability Index (SPADI) score. RESULTS: At 6 weeks, the total SPADI scores had improved significantly more in groups 1 and 2 compared with groups 3 and 4 (P = 0.0004). The total range of active and passive motion increased in all groups, with group 1 having significantly greater improvement than the other 3 groups. At 3 months, groups 1 and 2 still showed significantly greater improvement in SPADI scores than group 4. There was no difference between groups 3 and 4 at any of the followup assessments except for greater improvement in the range of shoulder flexion in group 3 at 3 months. At 12 months, all groups had improved to a similar degree with respect to all outcome measures. CONCLUSION: A single intraarticular injection of corticosteroid administered under fluoroscopy combined with a simple home exercise program is effective in improving shoulder pain and disability in patients with adhesive capsulitis. Adding supervised physiotherapy provides faster improvement in shoulder range of motion. When used alone, supervised physiotherapy is of limited efficacy in the management of adhesive capsulitis. PMID- 12632440 TI - Down-regulation of Th1-mediated pathology in experimental arthritis by stimulation of the Th2 arm of the immune response. AB - OBJECTIVE: To assess whether systemic administration of recombinant interferon gamma (rIFN gamma), a proinflammatory cytokine that influences the differentiation of naive T cells into Th1 cells, promotes the induction of arthritis in DBA/1 mice immunized with type II collagen (CII) in Freund's incomplete adjuvant (IFA) and to determine the antiarthritic effect of treatment with CII in IFA. METHODS: DBA/1 mice were immunized with CII in IFA and injected intraperitoneally with rIFN gamma (8,000 units/mouse/day) or with recombinant interleukin-12 (rIL-12; 100 ng/mouse/day). In another experiment, mice were immunized with CII in Freund's complete adjuvant (CFA) and treated with a single dose of CII in IFA on the day of immunization or on the day of disease onset. Mice were monitored to assess the effect of the treatment on the severity of disease. Th1/Th2 cytokines and anti-CII antibodies were measured by enzyme-linked immunosorbent assay. RESULTS: The administration of rIL-12 or rIFN gamma to mice immunized with CII in IFA restored the Th1 response and resulted in the development of arthritis. We then determined whether immunization with CII in IFA had the capacity to prevent and/or ameliorate collagen-induced arthritis. A single intraperitoneal injection of CII in IFA prevented arthritis when given at the time of immunization with CII in CFA and reduced disease severity when given at the time of arthritis onset. The administration of CII in IFA resulted in a profound down-regulation of IFN gamma production and an up-regulation of IL-10 in cultures of draining lymph node cells. CONCLUSION: These findings demonstrate that it is possible to deflect an ongoing pathogenic Th1 response to an antigen by reimmunization of the same antigen with a Th2-polarizing adjuvant. PMID- 12632441 TI - Placental immunomodulator ferritin, a novel immunoregulator, suppresses experimental arthritis. AB - OBJECTIVE: To determine the effect of treatment with C48, the recombinant cytokine-like domain of the novel human placental immunomodulator ferritin (PLIF) immunoregulator, on zymosan-induced arthritis (ZIA) in mice and on adjuvant induced arthritis (AIA) in rats. METHODS: The in vitro effect of PLIF/C48 was tested in mixed lymphocyte cultures (MLCs) of allogeneic mouse splenocytes. Arthritis was induced by intraarticular injection of zymosan into naive mice and by subcutaneous injection of Mycobacterium tuberculosis into rats. C48 was injected intraperitoneally daily from day 3 to day 9 or from day 7 to day 13 after induction of synovitis by zymosan, and every other day from day 2 to day 14 after induction of AIA. Swelling of the joints and histologic features of the synovium were assessed. Th1 and Th2 cytokines were quantified by enzyme-linked immunosorbent assay. RESULTS: Both PLIF and C48 significantly inhibited the in vitro immunoreactivity of mouse splenocytes in MLCs. Treatment of ZIA mice and AIA rats with C48 effectively reduced joint swelling. C48 treatment reduced synovial lining thickening, numbers of mononuclear cells and histiocytes, as well as cartilage destruction and bone erosions. In vitro, activated splenocytes from C48-treated ZIA and AIA animals produced significantly higher levels of interleukin-10 (IL-10). In animals with ZIA, this was accompanied by lower levels of tumor necrosis factor and IL-2. CONCLUSION: Human PLIF and C48 were shown to exert cross-species immunosuppressive activity in vitro. The in vivo suppression of articular inflammation in the experimental models of ZIA and AIA was the result of treatment with the antiinflammatory human C48. These results suggest that treatment with C48 may offer an effective immunotherapeutic means of controlling inflammatory polyarthritis. PMID- 12632442 TI - Association of common variation in glutathione S-transferase genes with premature development of cardiovascular disease in patients with systemic sclerosis. PMID- 12632444 TI - The diagnosis and prognosis of early arthritis: comment on the editorial by Scott. PMID- 12632446 TI - Anti-cyclic citrullinated peptide antibody in early rheumatoid arthritis: comment on the editorial by Scott. PMID- 12632447 TI - Fibroblast-like synoviocytes from rheumatoid arthritis patients express less FLICE-inhibitory protein than fibroblast-like synoviocytes from trauma patients: comment on the article by Schedel et al. PMID- 12632449 TI - The promise and limitations of DNA microarray analysis: comment on the editorial by Firestein and Pisetsky. PMID- 12632450 TI - Minimum information about a microarray experiment: comment on the editorial by Firestein and Pisetsky. PMID- 12632452 TI - Is decaf a decoy? Comment on the article by Mikuls et al. PMID- 12632453 TI - Dosage of hydroxychloroquine should be based on ideal body weight: comment on the letter by Alarcon. PMID- 12632454 TI - Dosage and mode of administration of methotrexate in clinical trials: comment on the article by Genovese et al. PMID- 12632456 TI - Multicentric reticulohistiocytosis treated successfully with an anti-tumor necrosis factor agent: comment on the article by Gorman et al. PMID- 12632457 TI - Acute adverse reaction to hylan G-F 20: comment on the article by Martens. PMID- 12632458 TI - HLA-B*2709 and spondylarthropathies: comment on the concise communication by Olivieri et al. PMID- 12632461 TI - Clinical image: hidden costs of anti-tumor necrosis factor alpha therapy. PMID- 12632463 TI - Human prefrontal and sensory cortical activity during divided attention tasks. AB - In our natural environment, the ability to divide attention is essential since we attend simultaneously to a number of sensory modalities, e.g., to visual and auditory stimuli. In this study, functional magnetic resonance imaging (fMRI) was used to study brain activation while a divided attention task was performed. Brain activation was also assessed under selective attention. Fourteen healthy male subjects aged between 19 and 28 years underwent fMRI studies using gradient EPI sequences. Cingulate activation was evident in all attention tasks. Focusing attention on one modality (visual or auditory) increased the activity in the corresponding primary and secondary sensory area. When attention is divided between both modalities, the activation in the sensory areas is decreased, possibly due to a limited capacity of the system for controlled processing. Left prefrontal activation, however, was evident selectively during the divided attention task. The present results suggest that this area may be important in the execution of controlled processing when attention is divided between two sources of information. These results support the view that the prefrontal cortex is involved in the central executive system and controls attention and information flow. PMID- 12632464 TI - Learning of tactile frequency discrimination in humans. AB - Learning is based on the remodeling of neural connections in the brain. The purpose of the present study was to examine the extent to which training-induced improvements in tactile frequency discrimination in humans are correlated with an increase of cortical representations in the primary somatosensory cortex. Healthy male subjects (n = 16) were trained in a tactile frequency discrimination task of the left ring finger. During the first 15 days of training, there was a steep improvement in frequency discrimination, which generalized from the trained finger to its homologue on the opposite hand, and to a lesser extent, to the other fingers on both hands. During the following 15 days of training, there was only a minor improvement in tactile frequency discrimination. Retention of improved performance in frequency discrimination 30 days after training was demonstrated for all digits. Cortical finger representation in the primary somatosensory cortex, as measured by magnetic source imaging, did not change during training. Because of the generalized training effect and the lack of detectable increase in the cortical field evoked from the trained finger, we assume that skill improvement was mediated predominantly by regions outside the primary somatosensory cortex. PMID- 12632465 TI - Functional somatotopy of finger representations in human primary motor cortex. AB - To assess the degree of fine-scale somatotopy within the hand area of the human primary motor cortex (M1), functional mapping of individual movements of all fingers was performed in healthy young subjects (n = 7) using MRI at 0.8 x 0.8 mm2 resolution and 4 mm section thickness. The experimental design comprised both a direct paradigm contrasting single digit movements vs. motor rest and multiple differential paradigms contrasting single digit movements vs. the movement of another digit. Direct mapping resulted in largely overlapping activations. A somatotopic arrangement was only recognizable when considering the mean center-of mass coordinates of individual digit representations averaged across subjects. In contrast, differential paradigms revealed more segregated and somatotopically ordered activations in single subjects. The use of center-of-mass coordinates yielded inter-digit distances ranging from 2.0 to 16.8 mm, which reached statistical significance for pairs of more distant digits. For the middle fingers, the functional somatotopy obtained by differential mapping was dependent on the choice of the digit used for control. These results confirm previous concepts that finger somatotopy in the human M1 hand area emerges as a functional predominance of individual digit representations sharing common areas in a distributed though ordered network. PMID- 12632466 TI - Independent component model for cognitive functions of multiple subjects using [15O]H2O PET images. AB - An independent component model of multiple subjects' positron emission tomography (PET) images is proposed to explore the overall functional components involved in a task and to explain subject specific variations of metabolic activities under altered experimental conditions utilizing the Independent component analysis (ICA) concept. As PET images represent time-compressed activities of several cognitive components, we derived a mathematical model to decompose functional components from cross-sectional images based on two fundamental hypotheses: (1) all subjects share basic functional components that are common to subjects and spatially independent of each other in relation to the given experimental task, and (2) all subjects share common functional components throughout tasks which are also spatially independent. The variations of hemodynamic activities according to subjects or tasks can be explained by the variations in the usage weight of the functional components. We investigated the plausibility of the model using serial cognitive experiments of simple object perception, object recognition, two-back working memory, and divided attention of a syntactic process. We found that the independent component model satisfactorily explained the functional components involved in the task and discuss here the application of ICA in multiple subjects' PET images to explore the functional association of brain activations. PMID- 12632467 TI - The importance of the dominant hemisphere in the organization of bimanual movements. AB - The successful control of upper limb movements is an essential skill of the human motor system. Yet, the neural organization of bimanual actions remains an issue of debate. Their control can be directed from both hemispheres, or, coordinated motion might be organized from the dominant (left) hemisphere. In order to unravel the neural mechanisms of bimanual behavior, we analyzed the standard task related and directed coherence between EEG signals picked up over the primary sensorimotor cortices in right-handed subjects during unimanual as well as bimanual in-phase (symmetrical) and anti-phase (asymmetrical) movements. The interhemispheric coherence in the beta frequency band (>13-30 Hz) was increased in both unimanual and bimanual patterns, compared to rest. During unimanual actions, the drive in the beta band from one primary sensorimotor cortex to the other was greater during movement of the contralateral as opposed to ipsilateral hand. In contrast, during bimanual actions, the drive from the dominant to the non-dominant primary sensorimotor cortex prevailed, unless task constraints induced by an external perturbation resulted in a substantial uncoupling of the hand movements, when interhemispheric coherence would also drop. Together, these results suggest that the contralateral hemisphere predominantly organizes unimanual movements, whereas coupled bimanual movements are mainly controlled from the dominant hemisphere. The close association between changes in interhemispheric coupling and behavioral performance indicates that synchronization of neural activity in the beta band is exploited for the control of goal-directed movement. PMID- 12632468 TI - White matter tractography using diffusion tensor deflection. AB - Diffusion tensor MRI provides unique directional diffusion information that can be used to estimate the patterns of white matter connectivity in the human brain. In this study, the behavior of an algorithm for white matter tractography is examined. The algorithm, called TEND, uses the entire diffusion tensor to deflect the estimated fiber trajectory. Simulations and imaging experiments on in vivo human brains were performed to investigate the behavior of the tractography algorithm. The simulations show that the deflection term is less sensitive than the major eigenvector to image noise. In the human brain imaging experiments, estimated tracts were generated in corpus callosum, corticospinal tract, internal capsule, corona radiata, superior longitudinal fasciculus, inferior longitudinal fasciculus, fronto-occipital fasciculus, and uncinate fasciculus. This approach is promising for mapping the organizational patterns of white matter in the human brain as well as mapping the relationship between major fiber trajectories and the location and extent of brain lesions. PMID- 12632469 TI - Comparison of various implicit solvent models in molecular dynamics simulations of immunoglobulin G light chain dimer. AB - The present study tests performance of different solvation models applied to molecular dynamics simulation of a large, dimeric protein molecule. Analytical Continuum Electrostatics (ACE) with two different parameter sets, older V98 and new V01, and Effective Energy Function (EEF) are employed in molecular dynamics simulation of immunoglobulin G (IgG) light chain dimer and variable domain of IgG light chain. Results are compared with explicit solvent and distance dependent dielectric constant (DDE) calculations. The overall analysis shows that the EEF method yields results comparable to explicit solvent simulations; however, the stability of simulations is lower. On the other hand, the ACE_V98 model does not seem to achieve the accuracy or stability expected in nanosecond timescale MD simulation for the studied systems. The ACE_V01 model greatly improves stability of the calculation; nonetheless, changes in radius of gyration and solvent accessible surface of the studied systems may indicate that the parameter set still needs to be improved if the method is supposed to be used for simulations of large, polymeric proteins. Additionally, electrostatic contribution to the solvation free energy calculated in the ACE model is compared with a numerical treatment of the dielectric continuum model. Wall clock time of all simulations is compared. It shows that EEF calculation is six times faster than corresponding ACE and 50 times faster than explicit solvent simulations. PMID- 12632470 TI - Concerning the reaction between singlet nitrenium ions and water: a computational investigation on competitive reaction paths. AB - The reaction between singlet nitrenium ions XNH(+) (X = F and Cl) and H(2)O has been investigated by high-level of theory ab initio calculations. The geometries of the involved intermediates, transition structures, and dissociation products have been optimized at the MP2(full)/6-31G(d) level of theory, and accurate total energies have been obtained using the Gaussian-3 (G3) procedure. The reaction commences by the exothermic formation of the F-NH-OH(2) (+) and Cl-NH-OH(2) (+) intermediates, which are in turn able to undergo two distinct low-energy reaction paths, namely, the isomerization to the N-protonated isomers of the hydroxylamines F-NH-OH or Cl-NH-OH, and the eventual extrusion of HF or HCl. The competitive or alternative occurrence of these two processes strictly depends on the nature of the substituent X. In the reaction between FNH(+) and H(2)O, the energy gained in the formation of the complex F-NH-OH(2) (+) from the association between FNH(+) and H(2)O, 52.1 kcal mol(-1), is by far larger than the activation barrier for the loss of HF from F-NH-OH(2) (+), computed as 24.9 kcal mol(-1). In addition, the F-NH-OH(2) (+) intermediate requires 33.0 kcal mol(-1) to overcome the barrier for the isomerization to F-NH(2)-OH(+). Therefore, the reaction between FNH(+) and H(2)O is expected to occur practically exclusively by HF elimination with formation of the HN-OH(+) ionic product. On the other hand, for the reaction between ClNH(+) and H(2)O, it is not possible to get a definitive conclusion on the competitive or alternative occurrence of the two reaction paths. In fact, the transition structure involved in the elimination of HCl from Cl-NH-OH(2) (+) is only 3.4 kcal mol(-1) lower in energy than the transition structure for the isomerization of Cl-NH-OH(2) (+) to Cl-NH(2)-OH(+). In addition, the absolute values of the energy barriers of these two processes, 24.2 and 27.6 kcal mol(-1), respectively, are comparable with the energy gained in the formation of the complex Cl-NH-OH(2) (+) from the association between ClNH(+) and H(2)O, 24.0 kcal mol(-).1 Therefore, the ClNH(+) cation is predicted to react with water significantly slower than FNH(+). PMID- 12632473 TI - Dual-level direct dynamics studies for the reactions of dimethyl ether with hydrogen atom and methyl radical. AB - The dual-level direct dynamics approach is employed to study the dynamics of the CH(3)OCH(3) + H (R1) and CH(3)OCH(3) + CH(3) (R2) reactions. Low-level calculations of the potential energy surface are carried out at the MP2/6 311+G(d,p) level of theory. High-level energetic information is obtained at the QCISD(T) level of theory with the 6-311+G(3df,3pd) basis set. The dynamics calculations are performed using variational transition state theory (VTST) with the interpolated single-point energies (ISPE) method, and small-curvature tunneling (SCT) is included. It is shown that the reaction of CH(3)OCH(3) with H (R1) may proceed much easier and with a lower barrier height than the reaction with CH(3) radical (R2). The calculated rate constants and activation energies are in good agreement with the experimental values. The calculated rate constants are fitted to k(R1) = 1.16 x 10(-19) T(3) exp(-1922/T) and k(R2) = 1.66 x 10(-28) T(5) exp(-3086/T) cm(3) mol(-1) s(-1) over a temperature range 207-2100 K. Furthermore, a small variational effect and large tunneling effect in the lower temperature range are found for the two reactions. PMID- 12632472 TI - Development of hardware accelerator for molecular dynamics simulations: a computation board that calculates nonbonded interactions in cooperation with fast multipole method. AB - Evaluation of long-range Coulombic interactions still represents a bottleneck in the molecular dynamics (MD) simulations of biological macromolecules. Despite the advent of sophisticated fast algorithms, such as the fast multipole method (FMM), accurate simulations still demand a great amount of computation time due to the accuracy/speed trade-off inherently involved in these algorithms. Unless higher order multipole expansions, which are extremely expensive to evaluate, are employed, a large amount of the execution time is still spent in directly calculating particle-particle interactions within the nearby region of each particle. To reduce this execution time for pair interactions, we developed a computation unit (board), called MD-Engine II, that calculates nonbonded pairwise interactions using a specially designed hardware. Four custom arithmetic processors and a processor for memory manipulation ("particle processor") are mounted on the computation board. The arithmetic processors are responsible for calculation of the pair interactions. The particle processor plays a central role in realizing efficient cooperation with the FMM. The results of a series of 50-ps MD simulations of a protein-water system (50,764 atoms) indicated that a more stringent setting of accuracy in FMM computation, compared with those previously reported, was required for accurate simulations over long time periods. Such a level of accuracy was efficiently achieved using the cooperative calculations of the FMM and MD-Engine II. On an Alpha 21264 PC, the FMM computation at a moderate but tolerable level of accuracy was accelerated by a factor of 16.0 using three boards. At a high level of accuracy, the cooperative calculation achieved a 22.7 fold acceleration over the corresponding conventional FMM calculation. In the cooperative calculations of the FMM and MD-Engine II, it was possible to achieve more accurate computation at a comparable execution time by incorporating larger nearby regions. PMID- 12632471 TI - Modeling zinc in biomolecules with the self consistent charge-density functional tight binding (SCC-DFTB) method: applications to structural and energetic analysis. AB - Parameters for the zinc ion have been developed in the self-consistent charge density functional tight-binding (SCC-DFTB) framework. The approach was tested against B3LYP calculations for a range of systems, including small molecules that contain the typical coordination environment of zinc in biological systems (cysteine, histidine, glutamic/aspartic acids, and water) and active site models for a number of enzymes such as alcohol dehydrogenase, carbonic anhydrase, and aminopeptidase. The SCC-DFTB approach reproduces structural and energetic properties rather reliably (e.g., total and relative ligand binding energies and deprotonation energies of ligands and barriers for zinc-assisted proton transfers), as compared with B3LYP/6-311+G** or MP2/6-311+G** calculations. PMID- 12632474 TI - Theoretical study concerning the reactivity of imine derivatives of polycyclic aromatic hydrocarbons. AB - The opening reaction of N-protonated polycyclic aromatic hydrocarbon imines has been computed by means of ab initio, density functional, and semiempirical methods of calculation. Imines are predicted to be more stable than the corresponding O-protonated derivatives, epoxides and diol epoxides. On the other hand, the activated N-methanesulfonylbenzene imine presented more favorable DeltaE( not equal ) and DeltaE(r) for ring opening due to the effect of hydrogen bond interactions. Anti and syn trans-diol benzene imines did not show a different behavior from the unsubstituted imine. According to these calculations, bay-region, fjord-region, and bay-region methyl-substituted compounds opened more easily among the imine derivatives, following the same reactivity pattern as the oxygen analogs. The exothermicity of the opening process correlated with the charge delocalization in the resulting carbocation. PMID- 12632475 TI - Vertical spectrum of the C2H2+ system. An open shell (SC)2-CAS-SDCI study. AB - The open shell (SC)(2)-CAS-SDCI method along with a basis set of atomic natural orbitals (ANO) has been applied for calculating the main ionization potentials of acetylene, as well as the manifold of excited states of the different symmetries up to 32 eV. In this method, the single and double excitations of a CAS space are generated and the corresponding CI matrix is corrected by means of the (SC)(2) procedure that cancels the size-extensivity error and adds some high order contributions. The mean absolute error for the outer-valence X (2)Pi(u)(1pi(u) ( 1)), A (2)Sigma(g) (+)(3sigma(g) (-1)), and B (2)Sigma(u) (+)(2sigma(u) (-1)) states, and the inner-valence C (2)Sigma(g) (+)(2sigma(g) (-1)) state is 0.1 eV. The excited states of C(2)H(2) (+) corresponding to the Sigma(g) (+), Sigma(u) (+), Pi(g), Pi(u), Delta(g), and Delta(u) symmetries are reported and their composition is discussed. The results are thoroughly compared to the best available multireference CI calculations. Recent multichannel CI results by Wells and Lucchese [J Chem Phys 1999, 110, 6365] have been used also as a guide for the discussion of the results. Discrepancies in the description of many multiconfigurational states by means of the (SC)(2)-CAS-SDCI wave function as compared to previous large MR-CI calculations are significant and are, consequently, remarked on. A large mixing of the 3sigma(g) (-1) and 2sigma(g) ( 1) processes is found in the A (2)Sigma(g) (+) and C (2)Sigma(g) (+) states, provided that the basis set is augmented with Rydberg functions. PMID- 12632476 TI - Sparse matrix multiplications for linear scaling electronic structure calculations in an atom-centered basis set using multiatom blocks. AB - A sparse matrix multiplication scheme with multiatom blocks is reported, a tool that can be very useful for developing linear-scaling methods with atom-centered basis functions. Compared to conventional element-by-element sparse matrix multiplication schemes, efficiency is gained by the use of the highly optimized basic linear algebra subroutines (BLAS). However, some sparsity is lost in the multiatom blocking scheme because these matrix blocks will in general contain negligible elements. As a result, an optimal block size that minimizes the CPU time by balancing these two effects is recovered. In calculations on linear alkanes, polyglycines, estane polymers, and water clusters the optimal block size is found to be between 40 and 100 basis functions, where about 55-75% of the machine peak performance was achieved on an IBM RS6000 workstation. In these calculations, the blocked sparse matrix multiplications can be 10 times faster than a standard element-by-element sparse matrix package. PMID- 12632477 TI - Performance assessment of density-functional methods for study of charge-transfer complexes. AB - Various density functionals are applied to a number of weakly bound intermolecular pi-pi charge-transfer (CT) complexes. Most functionals, including the recently developed mPWPW91 and mPW1PW91, grossly underestimate experimental excitation energies; good agreement is obtained only with the half-and-half hybrid BH&HLYP functional. PW91PW91 provides the best agreement with intermolecular distances measured in crystal, while the BH&HLYP values are about 0.1 A too long. Various hybrid functionals with nonlocal exchange correction provide binding energies that compare favorably with the experimental heats of formation measured in solution. PMID- 12632479 TI - Systematic errors in DFT calculations of haloalkane heats of formation. AB - The B3LYP and B3PW91 density functionals were employed with a large [BS1 = 6 311+G(3df,2p)] and small [BS2 = 6-311G(d,p)] basis set to compute enthalpies of formation (at optimized MP2/6-31G(d) geometries and with scaled HF/6-31G(d) frequencies) in the following series of haloalkanes: (1) the 15 fluoro-, chloro-, and chlorofluoromethanes, (2) the 18 fluorinated and chlorinated ethanes. Similar to earlier higher level calculations on haloalkanes, the computed enthalpies exhibited very large, systematic deviations from experiment. It was found that these errors could be largely eliminated using a very simple Bond Additivity Correction (BAC) formula, Delta(f)H degrees (BAC) = Delta(f)H degrees (calc) - n(CX). Delta(CX) [X = F, Cl], in which the BAC parameters, Delta(CF) and Delta(CCl) were determined by fitting the equation to experimental data on the four fluoroethanes and chloroethanes, respectively. The resultant BAC corrected enthalpies of formation are in excellent agreement with experiment, with RMS deviations in the same range as quoted RMS errors in measured enthalpies. Therefore, this simple BAC procedure may be utilized to provide reliable semiquantitative estimates of enthalpies of formation in larger haloalkanes, for which higher level ab initio calculations are not feasible. PMID- 12632478 TI - Molecular mechanics force field parameterization of the fluorescent probe rhodamine 6G using automated frequency matching. AB - Novel single-molecule fluorescence experimental techniques have prompted a growing need to develop refined computational models of dye-tagged biomolecules. As a necessary first step towards useful molecular simulations of fluorescence labeled biomolecules, we have derived a force field for the commonly used dye, rhodamine 6G (R6G). A novel automated method is used that includes fitting the molecular mechanics potential to both vibrational frequencies and eigenvector projections derived from quantum chemical calculations. The method is benchmarked on a series of aromatic molecules then applied to derive new parameters for R6G. The force field derived reproduces well the crystal structure of R6G. PMID- 12632480 TI - High-dimensional model representations generated from low order terms--lp-RS HDMR. AB - High-dimensional model representation (HDMR) is a general set of quantitative model assessment and analysis tools for improving the efficiency of deducing high dimensional input-output system behavior. RS-HDMR is a particular form of HDMR based on random sampling (RS) of the input variables. The component functions in an HDMR expansion are optimal choices tailored to the n-variate function f(x) being represented over the desired domain of the n-dimensional vector x. The high order terms (usually larger than second order, or equivalently beyond cooperativity between pairs of variables) in the expansion are often negligible. When it is necessary to go beyond the first and the second order RS-HDMR, this article introduces a modified low-order term product (lp)-RS-HDMR method to approximately represent the high-order RS-HDMR component functions as products of low-order functions. Using this method the high-order truncated RS-HDMR expansions may be constructed without directly computing the original high-order terms. The mathematical foundations of lp-RS-HDMR are presented along with an illustration of its utility in an atmospheric chemical kinetics model. PMID- 12632481 TI - nMoldyn: a program package for a neutron scattering oriented analysis of molecular dynamics simulations. AB - We present a new implementation of the program nMoldyn, which has been developed for the computation and decomposition of neutron scattering intensities from Molecular Dynamics trajectories (Comp. Phys. Commun 1995, 91, 191-214). The new implementation extends the functionality of the original version, provides a much more convenient user interface (both graphical/interactive and batch), and can be used as a tool set for implementing new analysis modules. This was made possible by the use of a high-level language, Python, and of modern object-oriented programming techniques. The quantities that can be calculated by nMoldyn are the mean-square displacement, the velocity autocorrelation function as well as its Fourier transform (the density of states) and its memory function, the angular velocity autocorrelation function and its Fourier transform, the reorientational correlation function, and several functions specific to neutron scattering: the coherent and incoherent intermediate scattering functions with their Fourier transforms, the memory function of the coherent scattering function, and the elastic incoherent structure factor. The possibility to compute memory function is a new and powerful feature that allows to relate simulation results to theoretical studies. PMID- 12632483 TI - Analysis of gene expression profile induced by EMP-1 in esophageal cancer cells using cDNA Microarray. AB - AIM: To obtain human esophageal cancer cell EC9706 stably expressed epithelial membrane protein-1 (EMP-1) with integrated eukaryotic plasmid harboring the open reading frame (ORF) of human EMP-1, and then to study the mechanism by which EMP 1 exerts its diverse cellular action on cell proliferation and altered gene profile by exploring the effect of EMP-1. METHODS: The authors first constructed pcDNA3.1/myc-his expression vector harboring the ORF of EMP-1 and then transfected it into human esophageal carcinoma cell line EC9706. The positive clones were analyzed by Western blot and RT-PCR. Moreover, the cell growth curve was observed and the cell cycle was checked by FACS technique. Using cDNA microarray technology, the authors compared the gene expression pattern in positive clones with control. To confirm the gene expression profile, semi quantitative RT-PCR was carried out for 4 of the randomly picked differentially expressed genes. For those differentially expressed genes, classification was performed according to their function and cellular component. RESULTS: Human EMP 1 gene can be stably expressed in EC9706 cell line transfected with human EMP-1. The authors found the cell growth decreased, among which S phase was arrested and G1 phase was prolonged in the transfected positive clones. By cDNA microarray analysis, 35 genes showed an over 2.0 fold change in expression level after transfection, with 28 genes being consistently up-regulated and 7 genes being down-regulated. Among the classified genes, almost half of the induced genes (13 out of 28 genes) were related to cell signaling, cell communication and particularly to adhesion. CONCLUSION: Overexpression of human EMP-1 gene can inhibit the proliferation of EC9706 cell with S phase arrested and G1 phase prolonged. The cDNA microarray analysis suggested that EMP-1 may be one of regulators involved in cell signaling, cell communication and adhesion regulators. PMID- 12632482 TI - Hepatocellular carcinoma with obstructive jaundice: diagnosis, treatment and prognosis. AB - Obstructive jaundice as the main clinical feature is uncommon in patients with hepatocellular carcinoma (HCC). Only 1-12 % of HCC patients manifest obstructive jaundice as the initial complaint. Such cases are clinically classified as "icteric type hepatoma", or "cholestatic type of HCC". Identification of this group of patients is important, because surgical treatment may be beneficial. HCC may involve the biliary tract in several different ways: tumor thrombosis, hemobilia, tumor compression, and diffuse tumor infiltration. Bile duct thrombosis (BDT) is one of the main causes for obstructive jaundice, and the previously reported incidence is 1.2-9 %. BDT might be benign, malignant, or a combination of both. Benign thrombi could be blood clots, pus, or sludge. Malignant thrombi could be primary intrabiliary malignant tumors, HCC with invasion to bile ducts, or metastatic cancer with bile duct invasion. The common clinical features of this type of HCC include: high level of serum AFP; history of cholangitis with dilation of intrahepatic bile duct; aggravating jaundice and rapidly developing into liver dysfunction. It is usually difficult to make diagnosis before operation, because of the low incidence rate, ignorant of this disease, and the difficulty for the imaging diagnosis to find the BDT preoperatively. Despite recent remarkable improvements in the imaging tools for diagnosis of HCC, such cases are still incorrectly diagnosed as cholangiocarcinoma or choledocholithiases. Ultrasonography (US) and CT are helpful in showing hepatic tumors and dilated intrahepatic and /or extrahepatic ducts containing dense material corresponding to tumor debris. Direct cholangiography including percutaneous transhepatic cholangiography (PTC) and endoscopic retrograde cholangiopancreatography (ERCP) remains the standard procedure to delineate the presence and level of biliary obstruction. Magnetic resonance cholangiopancreatography (MRCP) is superior to ERCP in interpreting the cause and depicting the anatomical extent of the perihilar obstructive jaundice, and is particularly distinctive in cases associated with tight biliary stenosis and along segmental biliary stricture. Choledochoscopy and bile duct brushing cytology could be alternative useful techniques in the differentiating obstructions due to intraluminal mass, infiltrating ductal lesions or extrinsic mass compression applicable before and after duct exploration. Jaundice is not necessarily a contraindication for surgery. Most patients will have satisfactory palliation and occasional cure if appropriate procedures are selected and carried out safely, which can result in long-term resolution of symptoms and occasional long-term survival. However, the prognosis of icteric type HCC is generally dismal, but is better than those HCC patients who have jaundice caused by hepatic insufficiency. PMID- 12632484 TI - Peptidergic innervation of human esophageal and cardiac carcinoma. AB - AIM: To investigate the distribution of neuropeptide-immunoreactive nerve fibers in esophageal and cardiac carcinoma as well as their relationship with tumor cells so as to explore if there is nerve innervation in esophageal and cardiac carcinoma. METHODS: Esophageal and cardiac carcinoma specimens were collected from surgical operation. One part of them were fixed immediately with 4 % paraformaldehyde and then cut with a cryostat into 40-microm-thick sections to perform immunohistochemical analysis. Antibodies of ten kinds of neuropeptide including calcitonin gene-related peptide (CGRP), galanin (GAL), substance P (SP), etc. were used for immunostaining of nerve fibers. The other part of the tumor specimens were cut into little blocks (1 mm(3)) and co-cultured with chick embryo dorsal root ganglia (DRG) to investigate if the tumor blocks could induce the neurons of DRG to extend processes, so as to probe into the possible reasons for the nerve fibers growing into tumors. RESULTS: Substantial amounts of neuropeptide including GAL-, NPY-, SP-immunoreactive nerve bundles and scattered nerve fibers were distributed in esophageal and cardiac carcinomas. The scattered nerve fibers waved their way among tumor cells and contacted with tumor cells closely. Some of them even encircled tumor cells. There were many varicosities aligned on the nerve fibers like beads. They were also closely related to tumor cells. In the co-culture group, about 63 % and 67 % of DRG co-cultured with esophageal and cardiac tumor blocks respectively extended enormous processes, especially on the side adjacent to the tumor, whereas in the control group (without tumor blocks), no processes grew out. CONCLUSION: Esophageal and cardiac carcinomas may be innervated by peptidergic nerve fibers, and they can induce neurons of DRG to extend processes in vitro. PMID- 12632485 TI - Expression of MUC1 in esophageal squamous-cell carcinoma and its relationship with prognosis of patients from Linzhou city, a high incidence area of northern China. AB - AIM: To further characterize the possible relationship between the molecular changes and prognosis of ESC and to elucidate the possible mechanisms involved. METHODS: 114 specimens of ESC were collected from Linzhou city, and all patients were followed up for more than 5 years after resection. Histopathological analysis and immunohistochemical staining (ABC) were employed to detect the alteration of MUC1. RESULTS: The positive immunostaining rate for MUC1 was 79 % (90/114), and the high-expression rate was 63 % (72/114). The mean survival periods (months) of those with high- and low-expression rates of MUC1 were 41 (95 % CI: 35, 47) and 52 (95 % CI: 45, 59), respectively. Patients in the low expression group obviously survived longer than those in high-expression group, and the difference was significant (P<0.05). The expression of MUC1 protein in the esophageal carcinoma specimens with metastasis was stronger than those without metastasis, the difference was also significant (P<0.05). The stepwise multivariate analysis showed that "differentiation", "expression of MUC1" and "TNM staging" were the most important factors affecting the prognosis of esophageal carcinoma patients (P<0.05). CONCLUSION: A good correlation between the alteration of MUC1 and the regional lymph node metastasis was observed. Furthermore, high-expression of MUC1 was associated with poor prognosis for esophageal cancer patients. These results indicated that MUC1 is a promising biomarker for predicting lymph node metastasis and prognosis in esophageal cancer. PMID- 12632486 TI - Resveratrol induces apoptosis in human esophageal carcinoma cells. AB - AIM: To investigate the apoptosis in esophageal cancer cells induced by resveratrol, and the relation between this apoptosis and expression of Bcl-2 and Bax. METHODS: In in vitro experiments, MTT assay was used to determine the cell growth inhibitory rate. Transmission electron microscope and TUNEL staining method were used to quantitatively and qualitively detect the apoptosis status of esophageal cancer cell line EC-9706 before and after the resveratrol treatment. Immunohistochemical staining was used to detect the expression of apoptosis regulated gene Bcl-2 and Bax. RESULTS: Resveratrol inhibited the growth of esophageal cancer cell line EC-9706 in a dose-and time-dependent manner. Resveratrol induced EC-9706 cells to undergo apoptosis with typically apoptotic characteristics, including morphological changes of chromatin condensation, chromatin crescent formation, nucleus fragmentation and apoptotic body formation. TUNEL assay showed that after the treatment of EC-9706 cells with resveratrol (10 mmol/L) for 24 to 96 hours, the AIs were apparently increased with treated time (P<0.05). Immunohistochemical staining showed that after the treatment of EC-9706 cells with resveratrol (10 mmol/L) for 24 to 96 hours, the PRs of Bcl-2 proteins were apparently reduced with treated time (P<0.05) and the PRs of Bax proteins were apparently increased with treated time (P<0.05). CONCLUSION: Resveratrol is able to induce the apoptosis in esophageal cancer. This apoptosis may be mediated by down-regulating the apoptosis-regulated gene Bcl-2 and up-regulating the expression of apoptosis-regulated gene bax. PMID- 12632488 TI - Difference of gene expression profiles between esophageal carcinoma and its pericancerous epithelium by gene chip. AB - AIM: To study the difference of gene expression between esophageal carcinoma and its pericancerous epithelium and to screen novel associated genes in the early stage of esophageal carcinogenesis by cDNA microarray. METHODS: Total RNA was extracted with the original single step way from esophageal carcinoma, its pericancerous epithelial tissue and normal esophageal epithelium far from the tumor. The cDNA retro-transcribed from equal quantity of mRNA was labeled with Cy5 and Cy3 fluorescence functioning as probes. The mixed probes were hybridized with two pieces of BioDoor 4 096 double dot human whole gene chip. Fluorescence signals were scanned by ScanArray 3 000 laser scanner and farther analyzed by ImaGene 3.0 software with the digital computer. RESULTS: (1) A total of 135 genes were screened out, in which 85 and 50 genes whose the gene expression levels (fluorescence intensity) in esophageal carcinoma were more than 2 times and less than 0.5 times respectively compared with the normal esophageal epithelium. (2) There were also total 31 genes, among then 27 and 4 whose expressions in pericancerous tissue were 2-fold up-regulated and 0.5-fold down-regulated respectively compared with normal esophageal epithelium. (3) There were 13 genes appeared simultaneously in both pericancerous epithelium and esophageal carcinoma, while another 18 genes existed in pericancerous epithelium only. CONCLUSION: With the parallel comparison among these three gene profiles, it was shown that (1). A total of 135 genes, Whose expression difference manifested as fluorescence intensity were more than 2 times between esophageal carcinoma and normal esophageal epithelium, were probably related to the occurrence and development of the esophageal carcinoma. (2). The 31 genes showing expression difference more than 2 times between pericancerous and normal esophageal epithelium might be relate to the promotion of esophageal pericancerosis and its progress. The present study illustrated that by using the gene chip to detect the difference of gene expression profiles might be of benefit to the gene diagnosis, treatment and prevention of esophageal carcinoma. PMID- 12632487 TI - HLA-DRB1 allele polymorphisms in genetic susceptibility to esophageal carcinoma. AB - AIM: To probe into the genetic susceptibility of HLA-DRB1 alleles to esophageal carcinoma in Han Chinese in Hubei Province. METHODS: HLA-DRB1 allele polymorphisms were typed by polymerase chain reaction with sequence-specific primers(PCR-SSP) in 42 unrelated patients with esophageal cancer and 136 unrelated normal control subjects and the associated HLA-DRB1 allele was measured by nucleotide sequence analysis with PCR.SAS software was used in statistics. RESULTS: Allele frequency (AF) of HLA-DRB1*0901 was significantly higher in esophageal carcinoma patients than that in the normal controls (0.2500 vs 0.1397, P=0.028, the odds ratio 2.053, etiologic fraction 0.1282). After analyzed the allele nucleotide sequence of HLA-DRB1*0901 which approachs to the corresponded exon 2 sequence of the allele in genebank. There was no association between patients and controls in the rested HLA-DRB1 alleles. CONCLUSION: HLA-DRB1*0901 allele is more common in the patients with esophageal carcinoma than in the healthy controls, which is positively associated with the patients of Hubei Han Chinese. Individuals carrying HLA-DRB1*0901 may be susceptible to esophageal carcinoma. PMID- 12632489 TI - Methylation and mutation analysis of p16 gene in gastric cancer. AB - AIM: To study methylation, frequencies of homozygous deletion and mutation of p16 gene in gastric carcinoma. METHODS: The methylation pattern in exon 1 and exon 2 of p16 gene was studied with polymerase chain reaction (PCR), using methylation sensitive restriction endonuclease HpaII and methylation insensitive restriction endonuclease MspI. PCR technique was used to detect homozygous deletions of exon 1 and exon 2 of p16 gene and single strand conformation polymorphism (SSCP) technique was used to detect the mutation of the gene. RESULTS: Hypermethylation changes in exon 1 and exon 2 of p16 gene were observed in 25 % and 45 % of 20 gastric cancer tissues, respectively, while no methylation abnormality was found in normal tissues. The homozygous deletion frequency of exon 1 and exon 2 of p16 gene in 20 gastric cancer tissues was 20 % and 10 %, respectively. No mutation was found in exon 1 of p16 gene, while abnormal single strands were found in 2 (10 %) cases in exon 2 as detected by SSCP. CONCLUSION: The results suggest that hypermethylation and abnormality of p16 gene may play a key role in the progress of gastric cancer. Hypermethylation of exon 2 of p16 gene may have effects on the carcinogenesis of gastric mucosa and may be a later event. PMID- 12632490 TI - Relationship between the expression of human telomerase reverse transcriptase gene and cell cycle regulators in gastric cancer and its significance. AB - AIM: To investigate the expression of human telomerase reverse transcriptase gene (hTRT) in gastric cancer (GC) and its relevance with cell cycle regulators including P16INK4, cyclin and P53. METHODS: In situ hybridization (ISH) for hTRT mRNA was performed in 53 cases of gastric cancer and adjacent cancerous tissues. Immunohistochemical staining (S-P method) for hTRT protein, P16INK4, cyclinD1 and P53 was performed in 53 cases of GC and adjacent cancerous tissues. RESULTS: Of 53 cases of GC, the expression of hTRT mRNA and hTRT protein was significantly higher than the expression of hTRT mRNA and hTRT protein in adjacent canerous tissues (P<0.01), the positive rates of hTRTmRNA and hTRT protein were 79.2 % and 88.6 %. There was a stastical difference of the expression of hTRT protein among well differentiated adenocarcinoma, poorly differentiated adenocarcinoma and mucoid carcinoma. And there was a highly significant positive correlation between the expression of hTRT mRNA and hTRT protein (r=0.625, P<0.01). However, the expression of hTRT mRNA and its protein in GC were not related with other clinicopathological parameters including gender, age, location and size of neoplasm, invasion depth, lymph node metastasis and clinical stage. There was a significant positive correlation between the expression of hTRT mRNA and cyclinD1 protein (r=0.350, P<0.01). There was a significant positive correlation between the expression of cyclinD1 protein and hTRT protein (r=0.549, P<0.01), so was between P53 and hTRT protein (r=0.319, P<0.05). CONCLUSION: The expression of hTRT gene is correlated significantly to the specific defects of cell cycle on G1/S check point; telomerase activity may depend on cell cycle in gastric cancer and it is available to clarify the molecular mechanism of telomerase activity regulation. The expression of hTRT mRNA and hTRT protein in GC is significantly different from the expression of hTRT mRNA and hTRT protein in adjacent cancerous tissue which indicates that these targets are correlated closely to the occurrence of GC and can provide important morphologic index for diagnosis of GC. PMID- 12632491 TI - Prospective cohort study of comprehensive prevention to gastric cancer. AB - AIM: To evaluate the preliminary effects of comprehensive prevention of gastric cancer in Zhuanghe County epidemiologically. METHODS: Stratified sampling and cluster sampling were applied to define the intervention group and the control group. The prospective cohort study was used for evaluating the effect of preventing gastric cancer. The relative risk (RR) and attributable risk percent (AR %) of intervention on gastric cancer death were calculated. Potential years of life lost (PLYY) of the disease was analyzed, and the RR and AR % of PYLL were calculated. Survival analysis was applied among the screened patients. RESULTS: In the first 4 years after intervening, the relative risk (RR) of intervention on death was 0.5059 (95 % CI: 0.3462-0.7392, P<0.05) with significance statistically. AR % of the intervention on death was 49.41 %. The RR of intervention on cumulative PYLL was 0.6778 (95 % CI: 0.5604-0.8198, P<0.05) with statistic significance. AR % of the intervention on cumulative PYLL was 30.32 %. The four-year survival rate of the screened patients was 0.6751 (95 % CI: 0.5298 0.9047). CONCLUSION: The initiative intervention results showed that the intervention approach used in the trial was effective, it reduced mortality and increased survival rate, and alleviated the adverse effect of gastric cancer on the health and life of screened population. PMID- 12632492 TI - Studies on microsatellite instability in p16 gene and expression of hMSH2 mRNA in human gastric cancer tissues. AB - AIM: To detect the loss of heterozygosity (LOH) frequency of microsatellite sites D9s171, D9s1604 of p16 gene and expression of hMSH2 mRNA in various differentiated types of gastric cancer, adjacent cancer tissues and normal gastric mucosa. METHODS: LOH was detected by polymerase chain reaction (PCR) denaturing polyacrylamide gel electrophoresis-silver staining. The expression of hMSH2 mRNA was examined with in situ hybridization. RESULTS: The frequency rate of LOH was significantly higher in gastric cancers than that in adjacent cancer tissues (P=0.032). No significant difference was noted among various differentiated types and various clinical stages of gastric cancers. The significantly reduced expression of hMSH2 mRNA positive signal cells exhibited in gastric cancers, in comparison with that in the adjacent cancer tissues and normal gastric mucosa, respectively (P=0.001). No significant difference was noted among various clinical stages of gastric cancers (P>0.05). The difference of positive signal cells in poorly differentiated cancers and those in well and moderately differentiated cancers were significant (P<0.001). CONCLUSION: The frequencies of LOH in two microsatellite sites, D9s171 and D9s1604, in p16 genome were associated with development of gastric cancer and no significant correlation was demonstrated between the LOH frequency and the cell differentiated types of tumor cells or clinical stages. There was a positive relationship between the expression of hMSH2 mRNA and the differentiated types of gastric cancer. PMID- 12632493 TI - Paclitaxel induces apoptosis in human gastric carcinoma cells. AB - AIM: To investigate the apoptosis in gastric cancer cells induced by paclitaxel, and the relation between this apoptosis and expression of Bcl-2 and Bax. METHODS: In in vitro experiments, MTT assay was used to determine the cell growth inhibitory rate. Transmission electron microscope and TUNEL staining method were used to quantitatively and qualitively detect the apoptosis status of gastric cancer cell line SGC-7901 before and after the paclitaxel treatment. Immunohistochemical staining was used to detect the expression of apoptosis regulated gene Bcl-2 and Bax. RESULTS: Paclitaxel inhibited the growth of gastric cancer cell line SGC-7901 in a dose-and time-dependent manner. Paclitaxel induced SGC-7901 cells to undergo apoptosis with typically apoptotic characteristics, including morphological changes of chromatin condensation, chromatin crescent formation, nucleus fragmentation and apoptotic body formation. Paclitaxel could reduce the expression of apoptosis-regulated gene Bcl-2, and improve the expression of apoptosis-regulated gene Bax. CONCLUSION: Paclitaxel is able to induce the apoptosis in gastric cancer. This apoptosis may be mediated by down expression of apoptosis-regulated gene Bcl-2 and up-expression of apoptosis regulated gene Bax. PMID- 12632494 TI - Protective effect of ascorbic acid in experimental gastric cancer: reduction of oxidative stress. AB - AIM: Oxidative stress participates in the cell carcinogenesis by inducing DNA mutations. Our aim was to assess whether ascorbic acid, an antioxidant, could have a role in preventing ROS (Reactive Oxygen Species) generation in experimental gastric carcinoma in a rat model. METHODS: Experimental gastric cancer was induced in twelve Wistar male rats (weighting 250-350 g) by profound duodeno-gastric reflux throught split gastrojenunostomy. The rats were allocated to the following groups: Group I (n=6) was the control; Group II (n=6) which was mantained with daily intake of tape water with Vitamin C (30 mg/Kg). After 6 or 12 months, samples of gastric tumor or non tumor mucosa were taken from the anastomosis of both groups. Oxidative stress was measured by superoxide quantification through lucigenin-amplified chemiluminescence base and by staining with Nitrobluetetrazolium. The histopathologic confirmation of adenocarcinoma was made by eosin-hemathoxilin method. RESULTS: The intestinal type of gastric adenocarcinoma was microscopically identified in all animals of group I whereas only 3 rats of group II showed an adenocarcinoma without macroscopic evidence of them. The cancers were located in the anastomosis in all cases. Basal luminescence from tumor gastric tissue generated 38.4+/-6.8 count per minute/mg/X10(6) (mean+/-SD) and 14.9+/-4.0 count per minute/mg/X10(6), respectively, in group I and II animals (P<0.05). The Nitrobluetetrazolium method showed intense staining in tumor tissues but not in non neoplasic mucosa. CONCLUSION: Experimental gastric tumors seem to produce more reactive oxygen species than non neoplasic gastric tissue. The reduction of oxidative stress and gastric tumor incidence in rats were induced by the intake of ascorbic acid. Therefore, it may have a role in the prevention of gastric carcinoma. PMID- 12632495 TI - The microcell mediated transfer of human chromosome 8 into highly metastatic rat liver cancer cell line C5F. AB - AIM: Our previous research on the surgical samples of primary liver cancer with CGH showed that the loss of human chromosome 8p had correlation with the metastatic phenotype of liver cancer. In order to seek the functional evidence that there could be a metastatsis suppressor gene(s) for liver cancer on human chromosome 8, we tried to transfer normal human chromosome 8 into rat liver cancer cell line C5F, which had high metastatic potential to lung. METHODS: Human chromosome 8 randomly marked with neo gene was introduced into C5F cell line by MMCT and positive microcell hybrids were screened by double selections of G418 and HAT. Single cell isolation cloning was applied to clone microcell hybrids. Finally, STS-PCR and WCP-FISH were used to confirm the introduction. RESULTS: Microcell hybrids resistant to HAT and G418 were obtained and 15 clones were obtained by single-cell isolation cloning. STS-PCR and WCP-FISH proved that human chromosome 8 had been successfully introduced into rat liver cancer cell line C5F. STS-PCR detected a random loss in the chromosome introduced and WCP-FISH found a consistent recombination of the introduced human chromosome with the rat chromosome. CONCLUSION: The successful introduction of human chromosome 8 into highly metastatic rat liver cancer cell line builds the basis for seeking functional evidence of a metastasis suppressor gene for liver cancer harboring on human chromosome 8 and its subsequent cloning. PMID- 12632497 TI - Detection of HBV, PCNA and GST-pi in hepatocellular carcinoma and chronic liver diseases. AB - AIM: To investigate the change of HBV DNA, PCNA and GST-pi in chronic liver disease and hepatocellular carcinoma (HCC). METHODS: Hepatitis B surface antigen (HBsAg), proliferating cell nuclear antigen (PCNA) and glutathione S-transferases (GST-pi) were detected by immunohistochemical staining and HBV DNA was detected by in situ hybridization (ISH) in formalin-fixed and paraffin-embedded sections with a total of 111 specimens of chronic hepatitis, liver cirrhosis, paratumorous tissue, HCC and normal liver tissue. RESULTS: The positive rates of HBsAg and HBVDNA were 62.5 %(15/24) and 75.0 %(12/16) in chronic hepatitis, 64.0 %(16/25) and 83.3 %(15/18) in liver cirrhosis, 72.7 %(16/22) and 85.7 %(12/14) in the paratumorous tissu and 45.0 %(14/31) and 64.3 %(9/14) in HCC. The positive HBVDNA granules in chronic hepatitis, liver cirrhosis and the paratumorous tissue were more intense than that in HCC. The positive rates of PCNA and GST-pi were 34.8 %(8/23) and 25.0 %(4/16) in chronic hepatitis, 73.7 %(14/19) and 17.6 %(3/17) in liver cirrhosis, 86.7 %(13/15) and 53.3 %(8/15) in the paratumorous tissue, 100 %(15/15) and 60.0 %(9/15) in HCC, respectively, and the positive rate of GST-pi in the paratumorous tissue was significantly higher than that in the liver cirrhosis without tumor (P<0.05), but same as that in HCC (P>0.05). CONCLUSION: The HBV infection may increase expression of PCNA and GST-pi. The paratumor cirrhosis may be a sequential lesion of precancerous cirrhosis around HCC. PMID- 12632496 TI - Effects of tachyplesin on the regulation of cell cycle in human hepatocarcinoma SMMC-7721 cells. AB - AIM: To investigate the effects of tachyplesin on the cell cycle regulation in human hepatcarcinoma cells. METHODS: Effects of tachyplesin on the cell cycle in human hepatocarcinoma SMMC-7721 cells were assayed with flow cytometry. The protein levels of p53, p16, cyclin D1 and CDK4 were assayed by immunocytochemistry. The mRNA levels of p21(WAF1/CIP1) and c-myc genes were examined with in situ hybridization assay. RESULTS: After tachyplesin treatment, the cell cycle arrested at G0/G1 phase, the protein levels of mutant p53, cyclin D1 and CDK4 and the mRNA level of c-myc gene were decreased, whereas the levels of p16 protein and p21(WAF1/CIP1) mRNA increased. CONCLUSION: Tachyplesin might arrest the cell at G0/G1 phase by upregulating the levels of p16 protein and p21(WAF1/CIP1) mRNA and downregulating the levels of mutant p53, cyclin D1 and CDK4 proteins and c-myc mRNA, and induce the differentiation of human hepatocacinoma cells. PMID- 12632498 TI - A novel HBV antisense RNA gene delivery system targeting hepatocellular carcinoma. AB - AIM: To construct a novel HBV antisense RNA delivery system targeting hapatocellular carcinoma and study its inhibitory effect in vitro and in vivo. METHODS: GE7,a 16-peptide specific to EGFR, and HA20,a homologue of N-terminus of haemagglutinin of influenza viral envelope protein, were synthesized and conjugated with polylysin. The above conjugates were organized into the pEBAF-as preS2, a hepatocarcinoma specific HBV antisense expression vector, to construct a novel HBV antisense RNA delivery system, named AFP-enhancing 4-element complex. Hepatocelluar carcinoma HepG2.2.15 cells was used to assay the in vitro inhibition of the complex on HBV. Expression of HBV antigen was assayed by ELISA. BALB/c nude mice bearing HepG2.2.15 cells were injected with AFP-enhancing 4 element complex. The expression of HBV antisense RNA was examined by RT-PCR and the size of tumor in nude mice were measured. RESULTS: The AFP-enhancing 4 element complex was constructed and DNA was completely trapped at the slot with no DNA migration when the ratio of polypeptide to plasmid was 1:1. The expression of HBsAg and HBeAg of HepG2.2.15 cells was greatly decreased after being transfected by AFP-enhancing 4-element complex. The inhibitory rates were 33.4 % and 58.5 % respectively. RT-PCR showed HBV antisense RNA expressed specifically in liver tumor cells of tumor-bearing nude mice. After 4 injections of AFP enhancing 4-element complex containing 0.2 micro g DNA, the diameter of the tumor was 0.995 cm+/-0.35, which was significantly smaller than that of the control groups(2.215 cm+/-0.25, P<0.05). CONCLUSION: AFP-enhancing 4-element complex could deliver HBV antisense RNA targeting on hepatocarcinoma and inhibit both HBV and liver tumor cells in vitro and in vivo. PMID- 12632499 TI - Is the expression of gamma-glutamyl transpeptidase messenger RNA an indicator of biological behavior in recurrent hepatocellular carcinoma? AB - AIM: To investigate the correlation between gamma-glutamyl transpeptidase (gamma GTP) expression in the primary HCC and post-resection recurrence and its biological behaviors. METHODS: Forty consecutive patients having curative resection for HCC were included in this study. The primers for reverse transcription polymerase chain reaction (RT-PCR) were corresponding to the 5' noncoding human gamma-GTP mRNA of fetal liver (type A), HepG2 cells (type B), and placenta (type C). Both the cancer and non-cancerous tissues of the resected liver were analyzed. The correlations between the expression of gamma-GTP and the clinicopathological variables and outcomes (recurrence and survival) were studied. RESULTS: Those with type B gamma-GTP mRNA in cancer had significant higher recurrence rate than those without it (63.6 % vs 14.3 %). Both those with type B in cancer and in non-cancer died significantly more than those without it (45.5 % vs 0 % and 53.6 % vs 0 %, respectively). By multivariate analysis, the significant predictors of recurrence included high serum AFP (P=0.0108), vascular permeation (P=0.0084), and type B gamma-GTP mRNA in non-cancerous liver (P=0.0107). The significant predictors of post-recurrence death included high serum AFP (P=0.0141), vascular permeation (P=0.0130), and daughter nodules (P=0.0053). As to the manifestations (recurrent number >=2, recurrent extent >=2 segments, extra-hepatic metastasis, and death) in recurrent patients, there were no statistical significant differences between those with type B in the primary tumor and those without it. The difference between those with type B in non cancerous liver and those without it also was not significant. CONCLUSION: Patients of HCC with type B gamma-GTP mRNA both in cancer and in non-cancerous tissue had a worse outcome, earlier recurrence, and more post-recurrence death. PMID- 12632500 TI - Hepatocyte transformation and tumor development induced by hepatitis C virus NS3 c-terminal deleted protein. AB - AIM: To study the effect of hepatitis C virus nonstructural protein 3 c-terminal deleted protein (HCV NS3-5') on hepatocyte transformation and tumor development. METHODS: QSG7701 cells were transfected with plasmid pRcHCNS3-5' (expressing HCV NS3 c-terminal deleted protein) by lipofectamine and selected in G418. The expression of HCV NS3 gene and protein was determined by PCR and immunohistochemistry respectively. Biological behavior of transfected cells was observed through cell proliferation assay, anchorage-independent growth and tumor development in nude mice. The expression of HCV NS3 and c-myc proteins in the induced tumor was evaluated by immunohistochemistry. RESULTS: HCV NS3 was strongly expressed in QSG7701 cells transfected with plasmid pRcHCNS3-5' and the positive signal was located in cytoplasm. Cell proliferation assay showed that the population doubling time in pRcHCNS3-5' transfected cells was much shorter than that in pRcCMV and non-transfected cells (24 h, 26 h, 28 h respectively). The cloning ratio of cells transfected with pRcHCNS3-5', pRcCMV and non transfected cells was 33 %, 1.46 %, 1.11 %, respectively, the former one was higher than that in the rest two groups (P<0.01). Tumor development was seen in nude mice inoculated with pRcHCNS3-5' transfected cells after 15 days. HE staining showed its feature of hepatocarcinoma, and immunohistochemistry confirmed the expressions of HCV NS3 and c-myc proteins in tumor tissue. The positive control group inoculated with HepG2 also showed tumor development, while no tumor developed in the nude mice injected with pRcCMV and non-transfected cells after 40 days. CONCLUSION: 1.HCV NS3 c-terminal deleted protein has transforming and oncogenic potential. 2. Human liver cell line QSG7701 may be used as a good model to study HCV NS3 pathogenesis. PMID- 12632501 TI - Antitumor immunopreventive and immunotherapeutic effect in mice induced by hybrid vaccine of dendritic cells and hepatocarcinoma in vivo. AB - AIM: To develop atumor vaccine by fusion of H22 hepatocarcinoma cells and DC, and to study its protective and therapeutical effect against H22 cell. METHODS: H22 DC vaccine was produced by PEG fusion of H22 and DC induced by cytokine released from splenic mononuclear cells, sorted by CD11c magnetic microbead marker. It was injected through the tail vein of the mice and the H(22)-DC oncogenesis was detected in the liver, spleen and lung. In order to study the therapeutical and protective effect of H(22)-DC against tumor H(22), two groups were divided: immune group and therapeutic group. Immune group was further divided into P, D, HD and H subgroups, immunized by PBS, DC, H(22)-DC and inactivated H(22), respectively, and attacked by H(22) cell. The tumor size, tumor weight, mice survival time and tumor latent period were recorded and statistically analyzed; Therapeutical group was divided into three subgroups of P, D and HD, and attacked by H(22), then treated with PBS, DC, and H(22)-DC, respectively. Pathology and flow cytometry were also applied to study the mechanism how the H(22)-DC vaccine attacked on the H(22) cell. RESULTS: 1. No oncogenesis was found in spleen, lung and liver after H22-DC injection. 2. Hybrid vaccine immunized mice had strongest CTL activity. 3. In the immune group, latent period was longer in HD subgroup than that in P, H and D subgroup; and tumor size and weight were smaller in HD subgroup than that in P, H and D subgroup. 4. In therapeutic group, tumor size was smaller in HD subgroup than that in P, D subgroup. CONCLUSION: 1. H22-DC tumor vaccine is safe without oncogenesis in vivo. 2. Hybrid vaccine can stimulate potent specific CTL activity against H22. 3. H22-DC vaccine has distinctive prophylatic effect on tumor H22 and can inhibit the tumor growth. PMID- 12632502 TI - Photodynamic inhibitory effects of three perylenequinones on human colorectal carcinoma cell line and primate embryonic stem cell line. AB - AIM: To investigate the photodynamic inhibitory effects of Elsinochrome A (EA), Hypocrellin A (HA) and Hypocrellin B (HB) on human colorectal carcinoma Hce-8693 cells and rhesus monkey embryonic stem R366.4 cells, via inducing apoptosis. METHODS: EA, HA and HB were extracted from metabolites of Hypomyces (Fr) Tul.Sp. R366.4 cells or Hce-8693 cells were cultured with different concentrations of EA, HA or HB respectively, irradiated and incubated with fresh medium for 2 h. Cell cycle analysis was performed by flow cytometry (FCM). Data were expressed as means +/-SD and analysis of variance and Student' t-test for individual comparisons. RESULTS: The photodynamic bioactivity of EA was first reported in this study. After irradiation for 5 min, 6 min, 10 min or 20 min, photoactivated EA at lower concentrations, which were 10(-7) Mol/L, 10(-6) Mol/L, 10(-5) Mol/L respectively, had no cytotoxic effects on R366.4 ES cells. Whereas, all of the three perylenequinones could induce apoptosis with a dose-dependent manner when Hce-8693 cells were incubated with photoactivated EA, HA and HB respectively. When Hce-8693 cells were incubated with EA at 10(-6) Mol/L and irradiated 5 min, 6 min, 10 min and 20 min respectively, the rates of EA-induced apoptosis were 0, 0, 13.4 % and 40.5 %. While the rates of HA-induced apoptosis were 29.5 %, 32.0 %, 40.2 % and 22.6 %. And the rates of HB-induced apoptosis were 0, 0, 0 and 13.7 % respectively. Meanwhile, after 10(-5) Mol/L treatment, the rates of EA-induced apoptosis were 32.7 %, 19.3 %, 26.4 % and 52.7 %, the rates of HA-induced apoptosis were 47.2 %, 39.1 %, 45.2 % and 56.6 %, and the rates of HB-induced apoptosis were 0, 0, 20.0 % and 13.9 % respectively. CONCLUSION: EA, HA and HB have significant anti-cancer activity. The order of photodynamic inhibitory effects on tumor cells would be approximately HA>EA>HB. The molecular mechanisms of apoptosis may not be induced by reactive oxygen species and are worth further investigation. PMID- 12632503 TI - Expression and significance of PTEN, hypoxia-inducible factor-1 alpha in colorectal adenoma and adenocarcinoma. AB - AIM: To investigate the expression and significance of PTEN, hypoxia-inducible factor-1 alpha (HIF-1alpha), and targeting gene VEGF during colorectal carciogenesis. METHODS: Total 71 cases colorectal neoplasms (9 cases of colorectal adenoma and 62 colorectal adenocarcinoma) were formalin fixed and paraffin-embedded, and all specimens were evaluated for PTEN mRNA, HIF-1alpha mRNA and VEGF protein expression. PTEN mRNA, HIF-1alpha mRNA were detected by in situ hybridization. VEGF protein was identified by citrate-microwave SP immunohistochemical method. RESULTS: There were significant differences in PTEN, HIF-1alpha and VEGF expression between colorectal adenomas and colorectal adenocarcinoma (P<0.05). The level of PTEN expression decreased as the pathologic stage increased. Conversely, HIF-1alpha and VEGF expression increased with the Dukes stage as follows: stage A (0.1029+/-0.0457: 0.1207+/-0.0436), stage B (0.1656+/-0.0329: 0.1572+/-0.0514), and stage C+D (0.2335+/-0.0748: 0.2219+/ 0.0803). For PTEN expression, there was a significant difference among Dukes stage A, B, and C+D, and the level of PTEN expression was found to be significant higher in Dukes stage A or B than that of Dukes stage C or D. For HIF-1alpha expression, there was a significant difference between Dukes stage A and B, and the level of HIF-1alpha expression was found to be significantly higher in Dukes stage C+D than that of Dukes stage A or B. The VEGF expression had similar results as HIF-1alpha expression. In colorectal adenocarcinoma, decreased levels of PTEN were significantly associated with increased expression of HIF-1alpha mRNA (r=-0.36, P<0.05) and VEGF protein (r=-0.48, P<0.05) respectively. The levels of HIF-1 were positively correlated with VEGF expression (r=0.71, P<0.01). CONCLUSION: Loss of PTEN expression and increased levels of HIF-1alpha and VEGF may play an important role in carcinogenesis and progression of colorectal adenocarcinoma. PMID- 12632504 TI - Application of autologous tumor cell vaccine and NDV vaccine in treatment of tumors of digestive tract. AB - AIM: To treat patients with stage I-IV malignant tumors of digestive tract using autologous tumor cell vaccine and NDV (Newcastle disease virus) vaccine, and observe the survival period and curative effect. METHODS: 335 patients with malignant tumors of digestive tract were treated with autologous tumor cell vaccine and NDV vaccine. The autologous tumor cell vaccine received were assigned for long-term survival observation. While these failed to obtain the autologous tumor tissue were given with NDV vaccine for a received short-term observation on curative effect. RESULTS: The colorectal cancer patients treated with autologous tumor cell vaccine were divided into two groups: the controlled group (subjected to resection alone) (n=257), the vaccine group (subjected to both resection and immunotherapy) (n=310). 25 patients treated with NDV immunotherapy were all at stage IV without having resection. In postoperation adjuvant therapy patients, the 5, 6 and 7-year survival rates were 66.51 %, 60.52 %, 56.50 % respectively; whereas in patients with resection alone, only 45.57 %, 44.76 % and 43.42 % respectively. The average survival period was 5.13 years (resection alone group 4.15 years), the median survival period was over 7 years (resection alone group 4.46 years). There were significant differences between the two groups. The patients treated with resection plus vaccine were measured delayed-type hypersensitivity (DTH) reactions after vaccination, (indurative scope >5 mm). The magnitude of DTH was related to the prognosis. The 5-year survival rate was 80 % for those with indurations greater than 5 mm, compared with 30 % for those with indurations less than 5 mm. The 1-year survival rate was 96 % for 25 patients treated with NDV immunotherapy. The total effective rate (CR+PR) was 24.00 % in NDV immunotherapy; complete remission (CR) in 1 case (4.00 %), partial remission (PR) in 5 cases (20.00 %), stabilizedin in 16 cases (64.00 %), progression (PD) in 1 case (4.00 %). After NDV vaccine immunotherapy, the number of NK cell increased and immune function imporved obviously. CONCLUSION: The autologous tumor cell vaccine and NDV vaccine can prolong the patients' life. NDV vaccine is notably effective for short-term with promotion of quality of life and can be used whenever necessary with good prospects. PMID- 12632505 TI - Full-length genome of wild-type hepatitis A virus (DL3) isolated in China. AB - AIM: To characterize the genome of an wild-type HAV isolate (DL3) in China. METHODS: A stool specimen was collected from hepatitis A patient from Dalian, China. HAV (DL3) was isolated and viral RNA was extracted. The genome of DL3 was amplified by reverse transcription and polymerase chain reaction (RT-PCR), followed by cloning into pGEM-T vector. The positive colonies were selected and sequenced. The full-length genome of DL3 was analyzed and compared with other wild-type HAV isolates. RESULTS: The genome of DL3 was 7 476 nucleotides (nt) in size, containing 732-nt 5'untranslated region (UTR), 6 681-nt open reading frame (ORF) which encoded a polyprotein of 2 227 amino acids (aa), and 63-nt 3'UTR. The base composition was 28.96 % A (2 165), 16.08 % C (1 202), 22.11 % G(1 653) and 32.85% U (2 456). Genomic comparisons with wild-type HAV isolates revealed that DL3 had the highest identity of 97.5 % for nt (185 differences) with AH1, the lowest identity of 85.7 % (1 066 differences) with SLF88. The highest identity of 99.2 % for amino acid (18 differences) appeared among DL3, AH2 and FH3, and the lowest identity of 96.8 % (72 differences) between DL3 and SLF88. Based upon comparisons of the VP1/2A junction and the VP1 amino terminus, DL3 was classified as subgenotype IA. Phylogenetic analysis showed that DL3 was closest to the isolates in Japan. CONCLUSION: The sequence comparison and phylogenetic analysis revealed that DL3 is most similar to the isolates in Japan, suggesting the epidemiological link of hepatitis A happened in China and Japan. PMID- 12632507 TI - A mutation specific polymerase chain reaction for detecting hepatitis B virus genome mutations at nt551. AB - AIM: The hepatitis B surface antigen (HBsAg) is considered to be one of the best markers for the diagnosis of acute and chronic HBV infection. But in some patients, this antigen cannot be detected by routine serological assays despite the presence of virus. One of the most important explanations for the lack of detectable HBsAg is that mutations which occur within the "a" determinant of HBV S gene can alter expression of HBsAg and lead to changes of antigenicity and immunogenicity of HBsAg accordingly. As a result, these mutants cannot be detected by diagnosis assays. Thus, it is essential to find out specific and sensitive methods to test the new mutants and further investigate their distribution. This study is to establish a method to investigate the distribution of the HBsAg mutant at nt551. METHODS: A mutation specific polymerase chain reaction (msPCR) was established for amplifying HBV DNA with a mutation at nt551. Four sets of primer pairs, P551A-PPS, P551G-PPS, P551C-PPS and P551T-PPS, with the same sequences except for one base at 3' terminus were designed and synthesized according to the known HBV genome sequences and the popular HBV subtypes, adr and adw, in China. At the basis of regular PCR method, we explored the specific conditions for amplifying HBV DNAs with a mutation at nt551 by regulating annealing temperature and the concentration of these primers. 126 serum samples from patients of hepatitis B were collected, among which 16 were positive for HBV S DNA in the nested PCR amplification. These 16 HBV S DNAs were detected by using the msPCR method. RESULTS: When the annealing temperature was raised to 71 degrees, nt551A and nt551G were amplified specifically by P551A-PPS and P551G-PPS; At 72 degrees and 5 pmole of the primers (each) in reaction of 25 microl volume, nt551C and nt551T were amplified specifically by P551C-PPS and P551T-PPS. 16 of HBV S gene fragments were characterized by using this method. 14 of them were positive for nt551A, one was positive for nt551G, and the other one was positive for nt551T. The results were confirmed by nucleotide sequencing. CONCLUSION: The mutation specific polymerase chain reaction is a specific and sensitive method for detecting the mutations of HBV genome at nt551. PMID- 12632508 TI - Expression of RNase H of human hepatitis B virus polymerase in Escherichia coli. AB - AIM: To amplify HBV-RNase H gene fragment and expression of RNase H for further use in the studies of HBV associated liver diseases. METHODS: The encoding gene of HBV-RNase H was separately amplified for the first half and second half (H1 and H2) by PCR from full length HBV gene and cloned into pT7Blue-T vector. Clones were first screened by digestion with XbaI and Hind III enzyme for the correct size, and analyzed further by DNA sequencing. The RNase H1 and H2 fragments isolated from XbaI and Hind III digestion products of pT7 Blue-RNase H plasmid were ligated to the GSTag expressing vectors separately, and expressed in E.coli BL21. The expressed proteins were checked by PAGE gel and Western blot. RESULTS: Both H1 and H2 nucleotide seqences consisting of known genes and proteins, in correct size, were further confirmed by Western blot to be the GST and RNase H1 or H2 fusion proteins. CONCLUSION: The successful cloning and expression of HBV RNase H will contribute to further research and application in HBV-associated diseases. PMID- 12632506 TI - Glucose intolerance in Chinese patients with chronic hepatitis C. AB - AIM: To investigate the prevalence and the risk factors of glucose intolerance in Chinese patients with chronic hepatitis C and to evaluate the relationship between interferon (IFN) treatment and glucose intolerance in these patients. METHODS: Prospective cross-sectional study was done to evaluate the prevalence of glucose intolerance in Chinese patients with chronic hepatitis C virus (HCV) infection from the outpatient clinic of Department of Family Medicine, Taipei Veterans General Hospital. Chronic hepatitis C was defined as persistent presence of anti-HCV and persistent elevation of liver transaminase for at least 1.5 folds for at least 6 months. Moreover, patients were further categorized into normal fasting glucose and glucose intolerance (diabetes mellitus (DM) and impaired fasting glucose) according to the diagnostic criteria of American Diabetic Association. RESULTS: Totally, 359 Chinese patients with chronic hepatitis C were enrolled (212 males and 147 females, mean age=58.1+/-13.0 years). One hundred and twenty-three patients (34.3 %) had received various forms of IFN treatment. One hundred and twenty-five patients (34.6 %) had glucose intolerance, including 99 patients (27.6 %) with DM and 26 patients (7.0 %) with impaired fasting glucose. In comparison with those with normal fasting glucose levels, patients with chronic hepatitis C with glucose intolerance were significantly older, had a significantly higher body mass index, and they were more likely to suffer from obesity, to have family history of diabetes and to have had previous IFN treatment. Stepwise multivariate logistic regression revealed significantly that age >=57 years, obesity, previous history of IFN treatment and the presence of family history of diabetes were independent risk factors associated with the presence of glucose intolerance in chronic hepatitis C patients. CONCLUSION: In conclusion, 34.6 % of Chinese patients with chronic hepatitis C had glucose intolerance. Chronic hepatitis C patients who were older in age, obese, had previous IFN treatment history and had family history of diabetes were prone to develop glucose intolerance. To our knowledge, this is the first population-based report to confirm that interferon treatment to be an independent risk factor to develop glucose intolerance. PMID- 12632510 TI - Overexpression of c-fos in Helicobacter pylori-induced gastric precancerosis of Mongolian gerbil. AB - AIM: To explore dysregulation of c-fos in several human malignancies, and to further investigate the role of c-fos in Helicobacter pylori (H. pylori)-induced gastric precancerosis. METHODS: Four-week-old male Mongolian gerbils were employed in the study. 0.5 mL 1X10(8) cfu/L suspension of H. pylori NCTC 11 637 in Brucella broth were inoculated orally into 20 Mongolian gerbils. Another 20 gerbils were inoculated with Brucella broth as controls. 10 of the infected gerbils and 10 of the non- infected control gerbils were sacrificed at 25 and 45 weeks after infection. The stomach of each gerbil was removed and opened for macroscopic observation. The expression of c-fos was analyzed by RT-PCR and immunohistochemical studies in H. pylori-induced gastric precancerosis of Mongolian gerbil. Half of each gastric antrum mucosa was dissected for RNA isolation and RT-PCR. beta-actin was used as the housekeeping gene and amplified with c-fos as contrast. PCR products of c-fos were analyzed by gel image system and the level of c-fos was reflected with the ratio of c-fos/beta-actin. The immunostaining for c-fos was conducted using monoclonal antibody of c-fos and the StreptAvidin-Biotin-enzyme Complex kit. RESULTS: H. pylori was constantly found in all infected animals in this study. After infection of H. pylori for 25 weeks, ulcers were observed in the antral and the body of stomach of 60 % infected animals (6/10). Histological examination showed that all animals developed severe inflammation, especially in the area close to ulcers, and multifocal lymphoid follicles appeared in the lamina propria and submucosa. After infection of H. pylori for 45 weeks, severe atrophic gastritis in all infected animals, intestinal metaplasia in 80 % infected animals (8/10) and dysplasia in 60 % infected animals (6/10) could be observed. C-fos mRNA levels were significantly higher after infection of H. pylori for 25 weeks (1.84+/-0.79), and for 45 weeks (1.59+/-0.37) than those in control-animals (0.74+/-0.22, P<0.01). C-fos mRNA levels were increased 2.5-fold by 25th week (P<0.01) and 2.1-fold by 45th week (P<0.01) in precancerosis induced by H. pylori, when compared with normal gastric epithelium of Mongolian gerbil. Immunohistochemical staining revealed exclusive nuclear staining of c-fos. Furthermore, there was a sequential increase in c-fos positive cells from normal epithelium to precancerosis. CONCLUSION: The study suggested that overexpression of c-fos occurs relatively early in gastric tumorigenesis in this precancerosis model induced by H. pylori. PMID- 12632509 TI - Virulence of water-induced coccoid Helicobacter pylori and its experimental infection in mice. AB - AIM: To explore the virulence and the infectivity of coccoid Helicobacter pylori (H. pylori) transformed from spiral form by exposure to sterile tap water. METHODS: Three strains of H. pylori, isolated from gastric biopsy specimens of confirmed peptic ulcer, were converted from spiral into coccoid form by exposure to sterile tap water. Both spiral and coccoid forms of H. pylori were tested for the urease activity, and the adherence to Hep-2 cells. The presence of flagella was examined under electron microscopy. In the experimental animal infection, the spiral and coccoid forms of H. pylori originated from the same strain F49 were inoculated intragastrically into BALB/c mice respectively four times at a 3-day interval. Half of the mice from each group were sacrificed at Day 21 and Day 28 after the last inoculation. Histology and H. pylori colonization were detected by urease test of gastric mucosa, cultures of H. pylori, and electron microscopy and so on. RESULTS: The urease activity and the ability of adherence to Hep-2 cells were found to be lower in coccoid H. pylori than that in its spiral form. For example, the transformation in strain F(44) led to a significant decrease of the adherence rate and adherence index from 70.0+/-5.3 % to 30.2+/-3.5 % (P<0.01), and from 2.6+/-0.4 to 0.86+/-0.3 (P<0.01), respectively. The flagella of coccoid H. pylori were observed under electron microscope. In the experimental infection in mice, the positive rate of gastric mucosa urease test was 93.8 % (15/16) in the group infected by spiral H. pylori and 50 % (8/16) in the group infected by coccoid H. pylori, and the estimated coccoid H. pylori colony number was 1.75 vs 0.56. The positive rates of H. pylori culture were 87.5 % (14/16) in spiral H. pylori group and 68.8 % (11/16) in coccoid H. pylori group. There was no significant difference in either urease test or bacterial culture rate between the groups examined at Day 21 and Day 28 after inoculation. Electron microscopic examination of the samples taken from both groups showed the adherence of H. pylori in spiral, bacillary and coccoid shapes to the epithelial cells of gastric wall. Histological examination showed the occurrence of gastric mucosal injury as indicated by various degrees of erosion, ulcer, and inflammatory cell infiltration. Mucosal injury was slighter in the mice infected by coccoid H. pylori. No positive result was obtained in the control group that received intragastrical administration of sterile tap water. CONCLUSION: Although the virulence of coccoid H. pylori induced by water decrease, coccoid H. pylori still remains a considerable urease activity and the adhering ability to epithelial cells. Furthermore, the flagella, an important component responsible for bacterial movement and infection, were still observed as a cellular structure of coccoid H. pylori under electron microscope. The coccoid H. pylori induced by water is capable of colonizing in gastric mucosa and causing gastrititis in mice. PMID- 12632511 TI - L-forms of H. pylori. AB - AIM: To study the occurrence of L-forms of H. pylori infection in patients with peptic ulcers and its association with possible changes of cellular immune function in the patients. METHODS: Endoscopic biopsy specimens of gastric antrum and gastric corpus were taken from 228 patients with peptic ulcers and inoculated into Skirrow selective medium for H. pylori vegetative forms and special medium for H. pylori L-forms, followed by bacterial isolation and identification. And peripheral venous blood of the patients was taken to detect the percentage of CD3(+), CD4(+) and CD8(+) with biotin-streptavidin (BSA) and the level of IL-2, IL-6 and IL-8 with ELISA. RESULTS: (1) The detection rates of H. pylori L-forms and vegetative forms in the patients were 50.88 % (116/228) and 64.91 % (148/228) respectively, and the co-infection rate of H. pylori L-forms and vegetative forms was 78.38 % (116/148). To be more exact, the detection rates of H. pylori L-forms in male and female patients were 57.04 % (77/135) and 41.94 % (39/93) respectively, and statistics found significant difference between them (P<0.05). Furthermore, the detection rates of H. pylori L-forms in patients aged 14 years-, 30 years-, 40 years- and 50 years- were 31.91 % (15/47), 42.86 % (24/56), 56.94 % (41/72) and 67.92 % (36/53) respectively, and there was significant difference between them (P<0.01). (2) The percentages of CD3(+), CD4(+), CD8(+), the ratio of CD4(+)/CD8(+), and the level of IL-2, IL-6, IL-8 in H. pylori-positive patients were (52.59+/-5.44) %, (35.51+/-5.74) %, (27.77+/-8.64) %, (1.56+/ 0.51), (2.66+/-0.47) mg/L, (108.62+/-5.85) ng/L and (115.79+/-7.18) ng/L respectively, compared with those in H. pylori-negative patients, the percentages of CD3(+), CD4(+) and the ratio of CD4(+)/CD8(+) decreased, but the level of IL 2, IL-6 increased, and the difference was significant (P<0.001-P<0.01). Moreover, the percentages of CD3(+), CD4(+), CD8(+), the ratio of CD4(+)/CD8(+), and the level of IL-2, IL-6, IL-8 in the patients with mixed infection of both H. pylori L-forms and vegetative forms were (51.69+/-5.28) %, (34.75+/-5.89) %, (27.15+/ 7.45) %, (1.48+/-0.47), (2.16+/-0.38) mg/L, (119.45+/-5.44) ng/L and (123.64+/ 6.24) ng/L respectively, compared with those in patients with simple infection of H. pylori vegetative forms, the percentage of CD4(+), the ratio of CD4(+)/CD8(+) and the level of IL-2 increased, but the level of IL-6 and IL-8 decreased, statistical difference was found between them (P<0.001-P<0.05). CONCLUSION: L forms variation often occurs in patients with peptic ulcers who are infected by H. pylori, which is commonly found in male patients and related to ages. The L forms variation of H. pylori can be an important factor causing disorder of cellular immune function in the patients with peptic ulcers who are infected by H. pylori. PMID- 12632512 TI - Inhibitive effect of cordyceps sinensis on experimental hepatic fibrosis and its possible mechanism. AB - AIM: To investigate the inhibitive effect and its possible mechanism of Cordyceps Sinensis (CS) on CCl(4)-plus ethanol-induced hepatic fibrogenesis in experimental rats. METHODS: Rats were randomly allocated into a normal control group, a model control group and a CS group. The latter two groups were administered with CCl(4) and ethanol solution at the beginning of the experiment to induce hepatic fibrosis. The CS group was also treated with CS 10 days after the beginning of CCl(4) and ethanol administration. All control groups were given corresponding placebo at the same time. At the end of the 9th week, rats in each group were humanely sacrificed. Blood and tissue specimens were taken. Biochemical, radioimmunological, immunohistochemical and molecular biological examinations were used to determine the level change of ALT, AST, HA, LN content in serum and TGFbeta(1), PDGF, collagen I and III expression in tissue at either protein or mRNA level or both of them. RESULTS: As compared with the model control group, serum ALT, AST, HA, and LN content levels were markedly dropped in CS group (86.0+/-34.4 vs 224.3+/-178.9, 146.7+/-60.2 vs 272.6+/-130.1, 202.0+/-79.3 vs 316.5+/-94.1 and 50.4+/-3.0 vs 59.7+/-9.8, respectively, P<0.05). Tissue expression of TGFbeta(1) and its mRNA, collagen I mRNA were also markedly decreased (0.2+/-0.14 vs 1.73+/-1.40, 1.68+/-0.47 vs 3.17+/-1.17, 1.10+/-0.84 vs 2.64+/-1.40, respectively, P<0.05). More dramatical drop could be seen in PDGF expression (0.87+/-0.43 vs 1.91+/-0.74, P<0.01). Although there was no statistical significance, it was still strongly suggested that collagen III mRNA expression was also decreased in CS group as compared with model control group (0.36+/-0.27 vs 0.95+/-0.65, P=0.0615). In this experiment, no significant change could be found in PDGF mRNA expression between two groups (0.35+/-0.34 vs 0.70+/ 0.46, P>0.05). CONCLUSION: Cordyceps sinensis could inhibit hepatic fibrogenesis derived from chronic liver injury, retard the development of cirrhosis, and notably ameliorate the liver function. Its possible mechanism involves inhibiting TGFbeta(1) expression, and thereby, down regulating PDGF expression, preventing HSC activation and deposition of procollagen I and III. PMID- 12632513 TI - Effect of cisapride on intestinal bacterial and endotoxin translocation in cirrhosis. AB - AIM: To investigate the effects of cisapride on intestinal bacterial overgrowth (IBO), bacterial and endotoxin translocation, intestinal transit and permeability in cirrhotic rats. METHODS: All animals were assessed with variables including bacterial and endotoxin translocation, intestinal bacterial overgrowth, intestinal transit and permeability. Bacterial translocation (BT) was assessed by bacterial culture of MLN, liver and spleen, IBO by a jejunal bacterial count of the specific organism, intestinal permeability by determination of the 24-hour urinary (99m)Tc-DTPA excretion and intestinal transit by measurement of the distribution of (51)Cr in the intestine. RESULTS: Bacterial translocation (BT) and IBO was found in 48 % and 80 % cirrhotic rats respectively and none in control rats. Urinary excretion of (99m)Tc-DTPA in cirrhotic rats with BT (22.2+/ 7.8) was greater than these without BT (10.5+/-2.9). Intestinal transit (geometric center ratio) was significantly delayed in cirrhotic rats (0.31+/ 0.06) and further more delayed in cirrhotic rats with BT (0.24+/-0.06) than these without BT (0.38+/-0.11). Cirrhotic rats with IBO had significantly higher rates of intestinal bacterial and endotoxin translocation, slower intestinal transit time and higher intestinal permeability than those without IBO. It was also found that BT was closely associated with IBO and the injury of intestinal barrier. Compared with the placebo group, cisapride-treated rats had lower rates of bacterial/endotoxin translocation and IBO, which was closely associated with increased intestinal transit and improved intestinal permeability by cisapride. CONCLUSION: These results indicate that endotoxin and bacterial translocation in cirrhotic rats may be attributed to IBO and increased intestinal permeability. Cisapride that accelerates intestinal transit and improve intestinal permeability might be helpful in preventing intestinal bacterial and endotoxin translocation. PMID- 12632514 TI - Effects of transmitters and interleukin-10 on rat hepatic fibrosis induced by CCl4. AB - AIM: To study the effects of transmitters ET, AgII, PGI(2), CGRP and GG on experimental rat hepatic fibrosis and the antifibrogenic effects of IL-10. METHODS: One hundred SD rats were randomly divided into 3 groups: control group (N): intraperitoneal injection with saline 2 ml.kg(-1) twice a week; the fibrogenesis group (C): intraperitoneal injection with 50 % CCl(4) 2 ml.kg(-1) twice a week; IL-10 treated group (E): besides same dosage of CCl(4) given, intraperitoneal injection with IL-10 4 ug.kg(-1) from the third week. In the fifth, the seventh and the ninth week, rats in three groups were selected randomly to collect plasma and liver tissues. The levels of ET, AgII, PGI(2), CGRP and GG were assayed by radioimmunoassay (RIA). The liver fibrosis was observed with silver staining. RESULTS: The hepatic fibrosis was developed with the increase of the injection frequency of CCl(4). The ET, AgII, PGI(2), CGRP and GG levels in serum of group N were 71.84+/-60.2 ng.L(-1), 76.21+/-33.3 ng/L, 313.03+/-101.71 ng/L, 61.97+/-21.4 ng/L and 33.62+/-14.37 ng/L, respectively; the levels of them in serum of group C were 523.30+/-129.3 ng/L, 127.24+/-50.0 ng/L, 648.91+/-357.29 ng/L, 127.15+/-62.0 ng/L and 85.26+/-51.83 ng/L, respectively; the levels of them in serum of group E were 452.52+/-99.5 ng/L, 90.60+/-44.7 ng/L, 475.57+/-179.70 ng/L, 102.2+/-29.7 ng/L and 38.05+/-19.94 ng/L, respectively. The histological examination showed that the degrees of the rats liver fibrosis in group E were lower than those in group C. CONCLUSION: The transmitters ET, AgII, PGI(2), CGRP and GG play a significant role in the rat hepatic fibrosis induced by CCl(4). IL-10 has the antagonistic action on these transmitters and can relieve the degree of the liver fibrosis. PMID- 12632515 TI - A selective tropism of transfused oval cells for liver. AB - AIM: To explore the biological behaviors of hepatic oval cells after transfused into the circulation of experimental animals. METHODS: Oval cells from male SD rat were transfused into the circulation of a female rat which were treated by a 2-AAF/CCl(4) program, through caudal vein. Sex-determining gene sry which located on Y chromosome was examined by PCR and in situ hybridization technique in liver, kidney and spleen of the experimental animals, respectively. RESULTS: The results of the cell-transplant experiment showed that the sry gene was detectable only in the liver but not in spleen and kidney of the experimental rats, and no signals could be detected in the control animals. It can be also morphologically proved that some exogenous cells had migrated into the parenchyma of the liver and settled there. CONCLUSION: The result means that there are exogenous cells located in the liver of the experimental animal and the localization is specific to the liver. This indicates that some "signal molecules" must exist in the circulation of the rats treated by 2-AAF/CCl(4). These "signal molecules" might play an important role in specific localization and differentiation of transfused oval cells. PMID- 12632516 TI - Comparative study in the effect of C-type natriuretic peptide on gastric motility in various animals. AB - AIM: To investigate the effect of natriuretic peptides on gastric motility in various animals, and the effect of C-type natriuretic peptide (CNP) on spontaneous contraction of gastric smooth muscle in rat, guinea-pig and human in vitro was compared. METHODS: Spontaneous contraction of gastric smooth muscle was recorded by four channel physiograph. RESULTS: In the guinea-pig and rat gastric antral circular smooth muscle, CNP markedly decreased the amplitude of spontaneous contraction but it didn't affect the frequency, however, the contractile activity was completely inhibited by CNP in gastric antral longitudinal smooth muscle. In the human gastric antral circular and longitudinal smooth musle, CNP completely inhibited spontaneous contraction. In the circular smooth muscle of guinea-pig and rat gastric fundus, CNP obviously decreased the amplitude of spontaneous contraction but it didn't affect the frequency, however, the contractile activity was completely inhibited by CNP in smooth muscle of fundus longitudinal. In the circular and longitudinal smooth muscle of guinea-pig gastric body, CNP at first induced a relaxation and then an increase in amplitude of spontaneous contraction (rebound contraction), but the frequency was not changed. After the circular smooth muscle of gastric body was pretreated with atropine, an M receptor blocker, the rebound contraction was abolished; In circular and longitudinal smooth muscle of rat gastric body, CNP induced a transient and slight relaxation and successively followed by the recovery in amplitude of spontaneous contraction but it also didn't affect the frequency. After the smooth muscle was pretreated with atropine, the transient and slight relaxation was replaced by long term and complete inhibition; The percentage of CNP-induced inhibition was 76.77+/-6.21 % (fundus), 67.21+/-5.32 % (body) and 58.23+/- 6.21 % (antral) in the gastric circular muscle, however, the inhibitory percentage was 100+/-0.00 % (fundus), 68.66+/- 3.55 % (body) and 100+/-0.00 % (antrum) in the gastric longitudinal smooth muscle of guinea-pigs; In the rat, the percentage of CNP-induced inhibition was 95.87+/-4.12 % (fundus), 94.91+/ 5.08 % (body) and 66.32+/-7.32 % (antrum)in the gastric circular smooth muscle, but in the longitudinal smooth muscle, CNP completely inhibited the spontaneous contraction. Using LY83583, a guanylate cyclase inhibitor, and zaparinast as a phosphoesterase inhibitor to inhibit the generation of cGMP, the effect of CNP on the spontaneous contraction was markedly weakened by LY83583, however, the inhibitory effect was enhanced by zaparinast. CONCLUSION: (1) CNP can obviously inhibit the spontaneous contraction of gastric antral circular and longitudinal smooth muscle in the rat, guinea-pig and human. The order of inhibitory potency is human >rat> guinea-pig. (2) In the same animals, the inhibitory effect of CNP on spontaneous contraction is the most powerful in fundus and the weakest in antrum, in the same position, the inhibitory effect on the circular smooth muscle is more powerful than that on longitudinal smooth muscle. (3) The inhibitory effect of CNP on spontaneous contraction in the gastric smooth muscle is mediated by a cGMP dependent pathway. PMID- 12632517 TI - Significance of changes of gastrointestinal peptides in blood and ileum of experimental spleen deficiency rats. AB - AIM: To explore the mechanism of spleen deficiency (SD) by studying the relationship of gastro-intestinal peptides level and ileal electro-mechanical activity of SD rats and cold restrain rats. METHODS: (1) spleen deficiency (SD) model was established by feeding Houpou:Zhishi: Dahuang in the ratio of 3:3:2, 3 ml/time, for 42 days. (2) The cold restrain stress model: Animals were restrained on grille and placed in a cool water at 18 degrees for 3 h. (3) Substance P (SP) and vasoactive intestinal peptide (VIP) levels in all layers of initial part of ileum and blood in rats were measured by radioimmunoassays (RIA) while changes of electric activity and motility in ileum of rats were recorded with electrode and strain gauge. RESULTS: SP levels in ileum and blood of experimental SD rats were significantly higher than that of the control groups (9.89+/-5.65 vs 1.22+/-1.18, P<0.005, in ileum; 22.7+/-3.95 vs 6.60+/-1.47, P<0.001, in blood) while the VIP levels of the SD rats were significantly lower than that of the controls (3.50+/ 2.01 vs 9.10+/-4.91, P<0.05, in ileum; 229.8+/-62.4 vs 560.4+/-151.3, P<0.001, in blood). As compared with the controls, the average frequency of slow electric waves (21.3+/-0.96 vs 18.2+/-2.28, P<0.05) and motility (21.5+/-0.58 vs 18+/ 2.65, P<0.005) of SD rats increased obviously and the frequency of fast waves of SD rat also increased. In spontaneous recovery cases, SP levels recovered significantly (compared with the SD groups, 2.99+/-0.62 vs 9.89+/-5.65, P<0.001, in ileum; 14.4+/-4.22 vs 22.7+/-3.95, P<0.001, in blood) but did not drop to normal. After the SD rats treated with Chinese herbs (Jiawei Sijun zi Tang), SP improved (compared with SD cases, 2.20+/-1.25 vs 9.89+/-5.65, (P<0.001), in ileum; 10.7+/-1.88 vs 22.7+/-3.95, (P<0.001), in blood) and VIP in blood also improved (compared with SD rats, 485.7+/-229.0 vs 229.8+/-62.4, P<0.01) while the amplitude of motility decreased apparently (compared with the SD rats, 0.64+/ 0.096 vs 0.89+/-0.15, P<0.01). The ileal SP levels of cool stress didn't change while the ileal VIP levels of cool stress became significantly lower than that of the control groups (2.87+/-0.87 vs 9.10+/-4.91, P<0.01). The blood SP levels of cool stress were significantly higher (15.60+/-1.83 vs 6.60+/-1.47, P<0.001) whereas the blood VIP levels of cool stress were significantly lower than that of the control group (153.4+/-70.46 vs 560.4+/-151.30, P<0.001). CONCLUSION: Changes of SP and VIP levels in initial part of ileum and blood of SD rats and cool stress rats may be closely related to the gastrointestinal motility disorders presented in SD and cool stress rats. the Chinese herbs (Jiawei Sijun zi Tang) currently used have partially therapeutic effect. PMID- 12632518 TI - Effect of pinaverium bromide on stress-induced colonic smooth muscle contractility disorder in rats. AB - AIM: To investigate the effect of pinaverium bromide, a L-type calcium channel blocker with selectivity for the gastrointestinal tract on contractile activity of colonic circular smooth muscle in normal or cold-restraint stressed rats and its possible mechanism. METHODS: Cold-restraint stress was conducted on rats to increase fecal pellets output. Each isolated colonic circular muscle strip was suspended in a tissue chamber containing warm oxygenated Tyrode-Ringer solution. The contractile response to ACh or KCl was measured isometrically on ink-writing recorder. Incubated muscle in different concentrations of pinaverium and the effects of pinaverium were investigated on ACh or KCl-induced contraction. Colon smooth muscle cells were cultured from rats and (Ca(2+))(i) was measured in cell suspension using the Ca(2+) fluorescent dye fura-2/AM. RESULTS: During stress, rats fecal pellet output increased 61 % (P<0.01). Stimulated with ACh or KCl, the muscle contractility was higher in stress than that in control. Pinaverium inhibited the increment of (Ca(2+))(i) and the muscle contraction in response to ACh or KCl in a dose dependent manner. A significant inhibition of pinaverium to ACh or KCl induced (Ca(2+))(i) increment was observed at 10(-6) mol/L. The IC(50) values for inhibition of ACh induced contraction for the stress and control group were 1.66X10(-6) mol/L and 0.91X10(-6) mol/L, respectively. The IC(50) values for inhibition of KCl induced contraction for the stress and control group were 8.13X10(-7) mol/L and 3.80X10(-7) mol/L, respectively. CONCLUSION: Increase in (Ca(2+))(i) of smooth muscle cells is directly related to the generation of contraction force in colon. L-type Ca(2+) channels represent the main route of Ca(2+) entry. Pinaverium inhibits the calcium influx through L-type channels; decreases the contractile response to many kinds of agonists and regulates the stress-induced colon hypermotility. PMID- 12632519 TI - Role of mitochondrial dysfunction in hydrogen peroxide-induced apoptosis of intestinal epithelial cells. AB - AIM: To study the role of mitochondrial dysfunction in hydrogen peroxide-induced apoptosis of intestinal epithelial cells. METHODS: Hydrogen peroxide-induced apoptosis of human intestinal epithelial cell line SW-480 was established. Cell apoptosis was determined by Annexin-V and PI double-stained flow cytometry and DNA gel electrophoresis. Morphological changes were examined with light and electron microscopy. For other observations, mitochondrial function, cytochrome c release, mitochondrial translocation and membrane potential were determined simultaneously. RESULTS: Percentage of apoptotic cells induced with 400 micro mol/L hydrogen peroxide increased significantly at l h or 3 h after stimulation and recovered rapidly. Meanwhile percentage of apoptotic cells induced with 4 mmol/L hydrogen peroxide increased with time. In accordance with these changes, we observed decreased mitochondrial function in 400 micro mol/L H2O2-stimualted cells at 1 h or 3 h and in 4 mmol/L H2O2-stimualted cells at times examined. Correspondingly, swelling cristae and vacuole-like mitochondria were noted. Release of cytochrome c, decreased mitochondrial membrane potential and mitochondrial translocation were also found to be the early signs of apoptosis. CONCLUSION: Dysfunctional mitochondria play a role in the apoptosis of SW-480 cell line induced by hydrogen peroxide. PMID- 12632520 TI - Combined therapy of allantoin, metronidazole, dexamethasone on the prevention of intra-abdominal adhesion in dogs and its quantitative analysis. AB - AIM: To observe the preventive effects of combined therapy of AMD (allantoin, metronidazolem and dexamethasone in combination) on intra-abdominal adhesion in dogs. METHODS: 20 dogs of both sexes were used in this study. After laparotomy under anesthesia, 2 cm section of cecal end was clamped and ligated, then 1 cm cecum section was cut and another 1 cm was kept. The cecum stump was closed with purse-string suture. Both parietal and visceral peritonea were stripped for an area of about 3X4 cm(2). Before the skin closure, the animals were divided into two groups randomly. The abdominal cavities in Group AMD (n=10) were rinsed by 200 ml of AMD solution, and with 50 ml left, whereas the control (n=10) received the equal volume of normal saline. After 7 d, the degree of intra-abdominal adhesions was evaluated by using the score method of ultrasonography and traditional dissection. RESULTS: Compared with the control, both the ultrasonography and traditional dissection scores in Group AMD were significantly decreased that marked as 2.0+/-1.25 vs 3.3+/-0.82 and 1.91+/-0.83 vs 3.3+/-0.82 respectively (P<0.01). CONCLUSION: The combined therapy of AMD is an effective way to prevent intra-abdominal adhesion, and ultrasonography is an useful tool to diagnose intra-abdominal adhesion. PMID- 12632521 TI - Preparation and identification of anti-transforming growth factor beta1 U1 small nuclear RNA chimeric ribozyme in vitro. AB - AIM: To study the preparation and cleavage activity of anti-transforming growth factor (TGF)beta1 U1 small nuclear (sn) RNA chimeric hammerhead ribozymes in vitro. METHODS: TGFbeta1 partial gene fragment was cloned into T-vector at the downstream of T7 promoter. (32)p-labeled TGFbeta1 partial transcripts as target RNA were transcribed in vitro and purified by denaturing polyacrylamide gel electrophoresis (PAGE). Anti-TGFbeta1 ribozymes were designed by computer, then synthetic ribozyme fragments were cloned into the U1 ribozyme vector pZeoU1EcoSpe containing U1 snRNA promoter/enhancer and terminator. (32)p-labeled U1 snRNA chimeric ribozyme transcripts were gel-purified, incubated with target-RNAs at different conditions and autoradiographed after running denaturing PAGE. RESULTS: Active U1snRNA chimeric ribozyme (U1Rz803) had the best cleavage activity at 50 degrees; at 37 degrees, it was active, K(m)=34.48 nmol/L, K(cat)=0.14 min(-1); while the point mutant ribozyme U1Rz803(m) had no cleavage activity, so these indicated the design of U1Rz803 was correct. CONCLUSION: U1Rz803 prepared in this study possessed the perfect specific catalytic cleavage activity. These results indicate U1 snRNA chimeric ribozyme U1Rz803 may suppress the expression of TGFbeta1 in vivo, therefore it may provide a new avenue for the treatment of liver fibrosis in the future. PMID- 12632523 TI - A mouse model of severe acute pancreatitis induced with caerulein and lipopolysaccharide. AB - AIM: To establish a non-traumatic, easy to induce and reproducible mouse model of severe acute pancreatitis (SAP) induced with caerulein and lipopolyasccharide (LPS). METHODS: Thirty-two healthy mature NIH female mice were selected and divided at random into four groups (each of 8 mice), i.e., the control group (NS group), the caerulein group (Cn group), the lipopolysaccharide group (LPS group), and the caerulein+LPS group (Cn+LPS group). Mice were injected intraperitoneally with caerulein only, or LPS only, and caerulein and LPS in combination. All the animals were then killed by neck dislocation three hours after the last intraperitoneal injection. The pancreas and exo-pancreatic organs were then carefully removed for microscopic examination. And the pancreatic acinus was further observed under transmission electron microscope (TEM). Pancreatic weight, serum amylase, serum nitric oxide (NO) concentration, superoxide dismutase (SOD) and malondialdehyde (MDA) concentration of the pancreas were assayed respectively. RESULTS: (1) NS animals displayed normal pancreatic structure both in the exocrine and endocrine. In the LPS group, the pancreas was slightly edematous, with the infiltration of a few inflammatory cells and the necrosis of the adjacent fat tissues. All the animals of the Cn group showed distinct signs of a mild edematous pancreatitis characterized by interstitial edema, infiltration of neutrophil and mononuclear cells, but without obvious parenchyma necrosis and hemorrhage. In contrast, the Cn+LPS group showed more diffuse focal areas of nonviable pancreatic and hemorrhage as well as systemic organ dysfunction. According to Schmidt's criteria, the pancreatic histologic score showed that there existed significant difference in the Cn+LPS group in the interstitial edema, inflammatory infiltration, parenchyma necrosis and parenchyma homorrhage in comparison with those of the Cn group, LPS group and NS group (P<0.01 or P<0.05). (2) The ultrasturcture of acinar cells was seriously damaged in the Cn+LPS group. Chromatin margination of nuclei was present, the number and volume of vacuoles greatly increased. Zymogen granules (ZGs) were greatly decreased in number and endoplasmic reticulum exhibited whorls. The swollen mitochondria appeared, the crista of which was decreased in number or disappeared. (3) Pancreatic weight and serum amylase levels in the Cn +LPS was significantly higher than those of the NS group and the LPS group respectively (P<0.01 or P<0.05). However, the pancreatic wet weight and serum amylase concentration showed no significant difference between the Cn+LPS group and the Cn group. (4) NO concentration in the Cn+LPS group was significantly higher than that of NS group, LPS group and Cn group(P<0.05 or P<0.01). (5) The SOD and MDA concentration of the pancreas in the Cn+LPS group were significantly higher than those of NS, LPS and Cn groups (P<0.05 or P<0.01). CONCLUSION: The mouse model of severe acute pancreatitis could be induced with caerulein and LPS, which could be non-traumatic and easy to induce, reproducible with the same pathological characteristics as those of SAP in human, and could be used in the research on the mechanism of human SAP. PMID- 12632522 TI - Analysis of spontaneous, gamma ray- and ethylnitrosourea-induced hprt mutants in HL-60 cells with multiplex PCR. AB - AIM: To explore the molecular spectra and mechanism of human hypoxanthine guanine phosphoribosyl transferase (hprt) gene mutation induced by ethyluitrosourea (ENU) and (60)Co gamma-rays. METHODS: Independent human promyelocytic leukemia cells (HL-60) mutants at the hprt locus were isolated from untreated, ethyluitrosourea (ENU) and (60)Co gamma-ray-exposed cells, respectively, and verified by two-way screening. The genetic changes underlying the mutation were determined by multiplex polymerase chain reaction (PCR) amplification and electrophoresis technique. RESULTS: With dosage increased, survival rate of plated cell reduced (in the group with dosage of ENU with 100-200 micro g/ml, P<0.01; in the group with dosage of (60)Co gamma-ray with 2-4 Gy, P<0.05) and mutational frequency increased (in the group of ENU 12.5-200.0 micro g/ml, P<0.05; in the group of (60)Co gamma-ray with 1-4 Gy, P<0.05) significantly. In the 13 spontaneous mutants analyzed, 92.3 % of mutant clones did not show any change in number or size of exon, a single exon was lost in 7.7 %, and no evidence indicated total gene deletion occurred in nine hprt exons. However, deletions were found in 79.7 % of ENU-induced mutations (62.5-89.4 %, P<0.01) and in 61.7 % of gamma-ray induced mutations (28.6-76.5 %, P<0.01). There were deletion mutations in all 9 exons of hprt gene and the most of induced mutations were chain deletion with multiplex exons (97.9 % in gamma-ray-induced mutants, 88.1 % in ENU-induced mutants). CONCLUSION: The spectra of spontaneous mutations differs completely from that induced by EUN or (60)Co gamma-ray. Although both ENU and gamma-ray can cause destruction of genetic structure, mechanism of mutagenesis between them may be different. PMID- 12632524 TI - In situ detection of TGF betas, TGF beta receptor II mRNA and telomerase activity in rat cholangiocarcinogenesis. AB - AIM: Initial report on the in situ examination of the mRNA expression of transforming growth factor betas (TGFbetas), TGFbeta type II receptor (TbetaRII) and telomerase activity in the experimental rat liver tissue during cholangiocarcinogenesis. METHODS: Rat liver cholangiocarcinogenesis was induced by 3'-methyl 4-dimethylazobenzene (3'Me-DAB). In situ hybridization was used to examine the TGFbetas) and TGFbeta type II receptor (TbetaRII) mRNA, in situ TRAP was used to check the telomerase activity in the tissue samples. RESULTS: There was no TGFbetas, TbetaRII mRNA expression or telomerase activity in the control rat cholangiocytes. The expression of TGFbeta1, TbetaRII was increased in regenerative, hyperplastic, dysplastic cholangiocytes and cholangiocarcinoma (CC) cells. The expression of TGFbeta2 mRNA was observed in only a part of hyperplastic, dysplastic cholangiocytes. TGFbeta3 expression was very weak, only in hyperplastic lesion. There was positive telomerase activity in the regenerative, hyperplastic, dysplastic cholangiocytes, and CC cells. Stroma fibroblasts of these lesions also showed positive TGFbetas, TbetaRII mRNA expression and telomerase activity. CONCLUSION: There were TGFbetas, TbetaRII expression and telomerase activity in hyperplastic, dysplastic cholangiocytes, cholangiocarcinoma cells as well as in stroma fibroblasts during cholangiocarcinogenesis. Their expression or activity is important in cholangiocarcinogenesis andstroma formation. PMID- 12632525 TI - Postoperative endoscopic surveillance of human living-donor small bowel transplantations. AB - AIM: To determine the significance of endoscopic surveillance in the diagnosis of acute rejection after human living-donor small bowel transplantations. METHODS: Endoscopic surveillance was performed through the ileostomy after human living donor small bowel transplantations. The intestinal mucosa was observed and biopsies were performed for pathological observations. RESULTS: Acute rejection was diagnosed in time by endoscopic surveillance. The endoscopic and pathological manifestations of acute rejection were described. CONCLUSION: Endoscopic surveillance and biopsy are reliable methods to diagnose the acute rejection after human living-donor small bowel transplantations. PMID- 12632526 TI - The effects of the formula of amino acids enriched BCAA on nutritional support in traumatic patients. AB - AIM: To investigate the formula of amino acid enriched BCAA on nutritional support in traumatic patients after operation. METHODS: 40 adult patients after moderate or large abdominal operations were enrolled in a prospective, randomly and single-blind-controlled study, and received total parenteral nutrition (TPN) with either formula of amino acid (AA group, 20 cases) or formula of amino acid enriched BCAA (BCAA group, 20 cases). From the second day after operation, total parenteral nutrition was infused to the patients in both groups with equal calorie and equal nitrogen by central or peripheral vein during more than 12 hours per day for 6 days. Meanwhile, nitrogen balance was assayed by collecting 24 hours urine for 6 days. The markers of protein metabolism were investigated such as amino acid patterns, levels of total protein, albumin, prealbumin, transferrin and fibronectin in serum. RESULTS: The positive nitrogen balance in BCAA group occurred two days earlier than that in AA group. The serum levels of total protein and albumin in BCAA group were increased more obviously than that in AA group. The concentration of valine was notably increased and the concentration of arginine was markedly decreased in BCAA group after the formula of amino acids enriched BCAA transfusion. CONCLUSION: The formula of amino acid enriched BCAA may normalize the levels of serum amino acids, reduce the proteolysis, increase the synthesis of protein, improve the nutritional status of traumatic patients after operation. PMID- 12632528 TI - Decision making in right-sided diverticulitis. AB - AIM: To evaluate systematically our nine-year experience in treating right-sided diverticulitis of the colon, and to explore its clinical and radiological relationship. METHODS: The clinical and radiological data of 40 patients with colonic diverticulitis treated in Mackay Memorial Hospital, Taipei, from 1993 through 2002 were reviewed retrospectively. RESULTS: The average age of the patients with right-sided diverticulitis was 53.1 years, which was 11.6 years younger than that of the patients with left-sided diverticulitis. The preoperative diagnosis of appendicitis was made in 8 of 13 right-sided diverticulitis patients. Nine (69 %) had right lower quadrant abdominal pain for more than 48 hours, and ten patients (77 %) presented with fever. CT findings suggesting acute right-sided diverticulitis including thickening of the intestinal wall and pericolonic inflammation were present in five patients. CONCLUSION: Right-sided diverticulitis is easily confused with acute appendicitis because it occurs at a somewhat younger age than that in left-sided diverticulitis. Barium enema and CT are helpful for the early diagnosis of right sided diverticulitis. While clearly not required in the majority of patients with right lower quadrant abdominal pain, barium enema and CT may be helpful in making the decision with a clinical history or physical examinations atypical of acute appendicitis. PMID- 12632527 TI - Adhesive small bowel obstruction: how long can patients tolerate conservative treatment? AB - AIM: To evaluate how long patients with small bowel obstruction caused by postoperative adhesions can tolerate conservative treatment. METHODS: The records of patients with small bowel obstruction due to postoperative adhesions were retrospectively reviewed. Data collected included the number of admissions, type of management for each admission, duration of conservative treatment, number of repeat laparotomies, and operative findings. RESULTS: One hundred fifty-five patients with this condition from January 1999 to December 2001, for a total of 293 admissions were enrolled in this study. Medical treatment alone was given in 220 admissions, and repeat laparotomy was performed in 73 admissions. The period of observation in patients managed medically ranged from 2 to 12 days (average: 6.9 days), while for those who underwent surgery, the range was 1 to 14 days (average 5.4 days). At surgery, adhesions were the only finding in 46 cases, while there were intestinal complications in 27, or 9.2 % of all 293 admissions. Fever and leukocytosis greater than 15 000/mm(3) were prediction of intestinal complications. CONCLUSION: With closely monitoring, most patients with small bowel obstruction due to postoperative adhesions could tolerate supportive treatment and recover well averagely within 1 week, although some patients require more than 10 days of observation. PMID- 12632529 TI - Esophageal ulceration complicating doxycycline therapy. AB - AIM: To report present state of iatrogenic drug-induced esophageal injury (DIEI) induced by medications in a private clinic. METHODS: Iatrogenic drug-induced esophageal injury (DIEI) induced by medications has been more frequently reported. In a private clinic we encountered 36 cases of esophageal ulcerations complicating doxycycline therapy in a mainly younger Saudi population (median age 29 years). RESULTS: The most frequent presenting symptoms were odynophagia, retrosternal burning pain and dysphagia (94 %, 75 % and 56 %, respectively). The diagnosis was according to medical history and confirmed by endoscopy in all patients. Beside withdrawal of doxycycline, when feasible, all patients were treated with a proton-pump inhibitor (PPI) and a prokinetic. Thirty patients who reported to the clinic after treatment were improved within 1-7 (median 1.7) days. CONCLUSION: Esophageal ulceration has to be suspected in younger patients with odynophagia, retrosternal burning pain and/or dysphagia during the treatment with doxycycline. PMID- 12632530 TI - Endosonography with linear array instead of endoscopic retrograde cholangiography as the diagnostic tool in patients with moderate suspicion of common bile duct stones. AB - AIM: To evaluate the diagnostic efficiency of endoscopic ultrasound (EUS) as the main imaging modality in patients with moderate suspicion of common bile duct stones (CBDS). METHODS: 55 patients with moderate clinical suspicion of CBDS were prospectively included to the study and evaluated with EUS. This study was done in single blind method in the clinical and biochemical data of patients. EUS was done with echo-endoscope Pentax FG 32-UA (f=5-7.5 MHz) and Hitachi EUB 405 ultrasound machine. Patients diagnosed with CBDS by EUS were excluded from this study and treated with ERC. All the other patients were included to the follow up study obtained by mail every 6 months for clinical evaluation (need of ERC or surgery). RESULTS: CBDS was found in 4 patients by EUS. Diagnosis was confirmed in all cases on ERC. The remaining 51 patients without CBDS on EUS were followed up for 6-26 months (meanly 13 months) There were: 40 women, 42 cholecystectomized patients, aged: 55 (mean). Biochemical values (mean values) were as follows: bilirubin: 14.9 micromol/L, alkaline phosphatase: 95 IU/L(-1), gamma-GTP: 131 IU/L(-1), ALT: 50 IU/L(-1), AST: 49 IU/L(-1)(-1). Only 1 patient was lost for follow up. In the remaining 50 patients with follow up, there was only 1 (2 %) patient with persistent biliary symptoms in whom CBDS was finally diagnosed by ERC with ES. All other patients remained symptoms free on follow up and did not require ERC or biliary surgery. CONCLUSION: Vast majority of patients with moderate suspicion of CBDS and no stones on EUS with linear array can avoid invasive evaluation of biliary tree with ERC. PMID- 12632531 TI - Crohn's disease and risk of fracture: does thyroid disease play a role? AB - AIM: To assess the role of thyroid disease as a risk for fractures in Crohn's patients. METHODS: A cross-sectional study was conducted from 1998 to 2000. The study group consisted of 210 patients with Crohn's disease. A group of 206 patients without inflammatory bowel disease served as controls. Primary outcome was thyroid disorder. Secondary outcomes included use of steroids, immunosuppressive medications, surgery and incidence of fracture. RESULTS: The prevalence of hyperthyroidism was similar in both groups. However, the prevalence of hypothyroidism was lower in Crohn's patients (3.8 % vs 8.2 %, P=0.05). Within the Crohn's group, the use of immunosuppressive agents (0 % vs 11 %), steroid usage (12.5 % vs 37 %), small bowel surgery (12.5 % vs 28 %) and large bowel surgery (12.5 % vs 27 %) were lower in the hypothyroid subset as compared to the euthyroid subset. Seven (3.4 %) Crohn's patients suffered fracture, all of whom were euthyroid. CONCLUSION: Thyroid disorder was not found to be associated with Crohn's disease and was not found to increase the risk for fractures. Therefore, screening for thyroid disease is not a necessary component in the management of Crohn's disease. PMID- 12632532 TI - Elevated serum values of procollagen III peptide (PIIIP) in patients with ulcerative colitis who will develop pseudopolyps. AB - AIM: To assess the impact of procollagen III peptide as a marker of collagenesis in the development of pseudopolyps in patients with ulcerative colitis. METHODS: Development of pseudopolyps was monitored in 25 patients with ulcerative colitis classified according to Powell-Tuck index as mild (n=12) or moderate (n=13) form of disease. Patients with a mild form of disease were treated with oral mesalazine medication (2-4 g/day) and local mesalazine preparation (suppository). Patients with a moderate form of disease received oral mesalazine medication (2-4 g/day), local mesalazine preparation (suppository) and local methylprednisolone at an initial dose of 60 mg/day, followed by dose tapering. How many significant variables (previously determined by analysis of variance) were elevated in the groups with and without pseudopolyp developement was observed. ROC analysis for calculation of new index was made. RESULTS: Serum values of procollagen III peptide (PIIIP), C-reactive protein (CRP) and C4 complement component (C4) were statistically significantly lower in the group of patients free from pseudopolyp development than those who developed one or more pseudopolyps (0.45+/-0.12 vs 1.42+/-0.70, P<0.0027; 7.6+/-4.7 vs 17.8+/-9.17, P<0.035; and 0.46+/-0.11 vs 0.34+/-0.16, P<0.068, respectively) at endoscopic conrtrols with patohistologically samples during 13 months. There were no statistically significant differences in the values of C3, ceruloplasmin and IgM between the two groups (P>0.05). Discrimination function analysis yielded highest standardized cannon coefficients for PIIIP (0.876), CRP (0.104), C3 (-0.534) and C4 (0.184) (P<0.036). The elevation in two of three laboratory variables (PIIIP, CRP and C4) reached sensitivity of 93 % and specificity of 90 % in the development of pseudopolyps. CONCLUSION: It is proposed that an increase in two of the three laboratory parameters (PIIIP, CRP and C4) could improve the accuracy of prediction of the development of pseudopolyps. When using PIIIP, CRP and C4 on decision making, the positive predictive value and accuracy were 90 % and 92 %, respectively. PMID- 12632533 TI - Synthesis of endotoxin receptor CD14 protein in Kupffer cells and its role in alcohol-induced liver disease. AB - AIM: To observe the synthesis of endotoxin receptor CD14 protein and its mRNA expression in Kupffer cells (KCs), and evaluate the role of CD14 in the pathogenesis of liver injury in rats with alcohol-induced liver disease (ALD). METHODS: Twenty-eight Wistar rats were divided into two groups: ethanol-fed group and control group. Ethanol-fed group was fed ethanol (dose of 5g-12 g/kg/d) and control group received dextrose instead of ethanol. Two groups were sacrificed at 4 wk and 8 wk, respectively. KCs were isolated and the synthesis of CD14 protein and its mRNA expression in KCs were determined by flow cytometric analysis (FCM) or the reverse transcription polymerase chain reaction (RT-PCR) analysis. The levels of plasma endotoxin and alanine transaminase (ALT) were measured by Limulus Amebocyte Lysate assay and standard enzymatic procedures respectively, and the levels of plasma tumor necosis factor (TNF)-alpha and interleukin (IL)-6 were both determined by ELISA. The liver pathology change was observed under light and electric microscopy. RESULTS: In ethanol-fed group, the percentages of FITC-CD14 positive cells were 76.23 % and 89.42 % at 4 wk and 8 wk, respectively. Compared with control group (4.45 % and 5.38 %), the difference was significant (P<0.05). The expressions of CD14 mRNA were 7.56+/-1.02 and 8.74+/-1.37 at 4 wk and 8 wk, respectively, which were significantly higher compared with the control group (1.77+/-0.21 and 1.98+/-0.23) (P<0.05). Plasma endotoxin levels at 4 wk and 8 wk increased significantly in ethanol-fed group (129+/-21 ng/L and 187+/-35 ng/L) than those in control rats (48+/-9 ng/L and 53+/-11 ng/L)(P<0.05). Mean values of plasma ALT levels increased dramatically in ethanol-fed rats (112+/-15 IU/L and 147+/-22 IU/L) than those in the control animals (31+/-12 IU/L and 33+/ 9 IU/L) (P<0.05). In ethanol-fed rats, the levels of TNF-alpha were 326+/-42 ng/L and 402+/-51 ng/L at 4 wk and 8 wk, respectively which were significantly higher than those in control group (86+/-12 ng/L and 97+/-13 ng/L) (P<0.05). The levels of IL-6 were 387+/-46 ng/L and 413+/-51 ng/L, which were also higher than control group (78+/-11 ng/Land 73+/-10 ng/L) (P<0.05). In liver section from ethanol-fed rats, there were marked pathological changes including steatosis, cell infiltration and necrosis. No marked pathological changes were seen in control group. CONCLUSION: Ethanol administration led to a significant synthesis of endotoxin receptor CD14 protein and its gene expression in KCs, which maybe result in the pathological changes of liver tissue and hepatic functional damages. PMID- 12632534 TI - Concomitant hepatocellular adenoma and adenomatous hyperplasia in a patient without cirrhosis. AB - AIM: Hepatocelluar adenoma (HCA) and adenomatous hyperplasia (AH) are rare benign tumors of the liver. HCA is usually found in women who use oral contraceptives. AH usually occurs in patients with liver cirrhosis. Both tumors have potential for malignant transformation. METHODS: We described a male adult with chronic liver disease (CLD) who had been known to be a hepatitis B carrier (HBV) for years. He was found to have a space-occupying lesion with a suspicion of hepatocellular carcinoma (HCC) by abdominal ultrasonography. His alpha fetoprotein (AFP) was normal. Angiographic findings were consistent with the diagnosis of HCC, he wished to avoid an operation, was treated with transcatheter hepatic arterial embolization. RESULTS: He subsequently consented to surgery, and a right lobectomy was performed. The liver pathology disclosed HCA with nuclear dysplasia and post-embolization effects. In addition, there were multiple small foci of AH with nuclear dysplasia in the resected liver. Although he had some focal areas of cirrhosis-like change or post-embolization effect, the AH was associated only with normal liver tissue. CONCLUSION: This case confirms that HCA and AH may resemble HCC on imaging studies, and that AH may occur in CLD in the absence of cirrhotic change. PMID- 12632535 TI - Successful rescuing a pregnant woman with severe hepatitis E infection and postpartum massive hemorrhage. AB - AIM: To sum up the experience of the successful therapy for the severe hepatitis of pregnant woman with postpartum massive hemorrhage. METHODS: The advanced therapeutic methods including the bilateral uterine artery embolism, hemodialysis and artificial liver support therapy were performed with comprehensive medical treatments and the course of the successful rescuing the patient was analyzed. RESULTS: Through the hospitalization of about two mouths the patient and her neonatus had gotten the best of care in our department and pediatric department separately. Both of them were discharged in good condition. CONCLUSION: The key points for a successful therapy of the pregnant woman with severe hepatitis are termination of the pregnancy and the control of their various complications. It was suggested that the proper combination of these measures of modern therapy would race against time for renewing of hepatic and renal functions. PMID- 12632537 TI - Evidence speaks for itself. PMID- 12632536 TI - Large pedinculated antral hyperplastic gastric polyp traversed the bulbus causing outlet obstruction and iron deficiency anemia: endoscopic removal. AB - We present here a large (3 cm) hyperplastic gastric polyp prolapsed into duodenum and caused outlet obstruction and iron deficiency anemia in 60 years old male patient. Endoscopic removal was performed successfully. PMID- 12632538 TI - Alternative cancer therapies can be dangerous. "Natural" and "herbal" are not synonymous with "safe". PMID- 12632539 TI - Addressing A-fib: rhythm or rate? For many, rate control may be preferable to rhythm control. PMID- 12632540 TI - Say "nuts" to diabetes? Nuts may reduce your risk of diabetes, but don't overdo it. PMID- 12632541 TI - Dealing with drug advertising. The FDA can't always enforce the rules on misleading ads. PMID- 12632542 TI - Acne cream can reverse sun damage. PMID- 12632543 TI - Acetaminophen is main cause of acute liver failure. PMID- 12632545 TI - New drug treats MS, Crohn's disease. PMID- 12632544 TI - Acrylamide: are fears bona fide? PMID- 12632546 TI - Benign breast-tumor freeze-out. PMID- 12632548 TI - Finding foot-friendly shoes. PMID- 12632547 TI - Diuretics: first choice for treating high BP. PMID- 12632549 TI - Human studies are more valuable than test-tube science. PMID- 12632550 TI - I'm a 65-year-old man with high blood pressure and a family history of stroke. Which stroke-prevention medication would be better for me--aspirin or clopidogrel (Plavix)? PMID- 12632551 TI - What is pulmonary fibrosis, and how is it treated? PMID- 12632552 TI - Getting a hand from social agencies. PMID- 12632554 TI - Body image. How you see yourself. PMID- 12632553 TI - Diabetes medicine update. Old dogs, new tricks. PMID- 12632555 TI - Diabetes quiz. How much do you know about smoking? PMID- 12632556 TI - Eat well, spend less. PMID- 12632558 TI - Choosing dental care. PMID- 12632557 TI - Wound healing for foot ulcers. PMID- 12632559 TI - Cardiac rehabilitation. PMID- 12632560 TI - What you should know about celiac disease. PMID- 12632561 TI - For parents. When more than one family member has diabetes. PMID- 12632562 TI - Sports and energy drinks. PMID- 12632563 TI - Erosion from filtered water. PMID- 12632564 TI - Toothache from composite filling. PMID- 12632565 TI - Need for alarm calls. PMID- 12632566 TI - Counselling, psychological morbidity and termination of pregnancy. PMID- 12632567 TI - Spotlights;research samplings; literatures, politics, photography and athletes. PMID- 12632568 TI - Bibliography. Current world literature. Cataract surgery and lens implantation. PMID- 12632569 TI - Medical Artists' Association of Great Britain 53rd Annual Conference, University of Cambridge, April 24-26, 2002. PMID- 12632570 TI - Involvement of nitric oxide/cyclic GMP signaling pathway in the regulation of fatty acid metabolism in rat hepatocytes. AB - The role of nitric oxide (NO)/guanosine 3',5'-cyclic monophosphate (cGMP) signaling pathway in the regulation of fatty acid metabolism was investigated in rat hepatocytes. Treatment with NO donors, which are known to activate soluble guanylyl cyclase, inhibited in parallel fatty acid synthesis de novo and acetyl CoA carboxylase activity. This effect was mimicked by 8-Br-cGMP and abolished by KT5823, a selective inhibitor of cGMP-dependent protein kinase (PKG). Furthermore, specific and hydrolysis-resistant activators of PKG, and inhibitors of Ca2+ release from endoplasmic reticulum, were also effective in inhibiting both fatty acid-synthesizing activities. These results suggest that this biological action of NO is regulated by a signaling cascade involving soluble guanylyl cyclase, cGMP, and PKG, and may be mediated, at least in part, by inhibition of Ca2+ release from endoplasmic reticulum. In addition, 8-Br-cGMP was able to stimulate fatty acid oxidation by two different mechanisms: the relieving of malonyl-CoA-dependent inhibition by lowering levels of this product of acetyl CoA carboxylase, and a malonyl-CoA-independent stimulation of carnitine palmitoyltransferase I. Taken together, results of this study suggest that NO/cGMP signaling pathway is endowed with regulatory properties in fatty acid metabolism, and may have a physiological role in the control of this metabolism in liver. PMID- 12632571 TI - Rethinking coma cocktails. PMID- 12632573 TI - [Askin's tumour]. PMID- 12632574 TI - [An atypical case of Demons-Meigs syndrome]. PMID- 12632576 TI - Inhalation injury: diagnosis. PMID- 12632577 TI - Update on ATAC. PMID- 12632578 TI - [Dragan Dugalic 1992-2000]. PMID- 12632579 TI - Informing the consumer. PMID- 12632580 TI - King Faisal International Prize awarded. PMID- 12632581 TI - Cancer drugs: the next 10 years. PMID- 12632582 TI - Research notes. The role and function of physician executives. PMID- 12632583 TI - [Radiology and the law. Reimbursement also for missed appointment]. PMID- 12632584 TI - [Status of medical physics in diagnostic radiology and nuclear medicine]. PMID- 12632585 TI - ["Method-Oriented Basic Research" Working Group]. PMID- 12632586 TI - [Imaging with Heart. 19 October 2002, AKH Vienna]. PMID- 12632587 TI - Pathophysiology and clinical implications of perioperative fluid excess. PMID- 12632588 TI - [Regarding the title picture: McCune-Albright syndrome. Synonym: Albright-McCune Sternberg syndrome]. PMID- 12632589 TI - Mass health education on sex and sexuality and its impact on Cuba: an interview based medical report on the development, evolution, and current status of this educational program in Cuba, with emphasis on women's health. PMID- 12632590 TI - Patentability and higher life forms. PMID- 12632591 TI - Why are women missing from outpatient cardiac rehabilitation programs? A review of multilevel factors affecting referral, enrollment, and completion. AB - OBJECTIVES: The objectives of this paper are to (1) systematically review the evidence for patient, provider, and programmatic factors that may influence women's referral to, enrollment in, and completion of outpatient cardiac rehabilitation and (2) make empirically based recommendations for future women's health research. METHODS: Using a defined inclusion/exclusion criteria, this review involved a systematic review and description analysis of the published peer-review literature. RESULTS: The review yielded 23 studies described in 25 publications. Although gaps in the knowledge base exist and several methodological concerns limit the evidence, this body of work suggests that age, personal resources, low rates of physician referral, and weak recommendations to participate in rehabilitation may explain why women are missing from this life saving intervention. CONCLUSIONS: Practitioners engaged in the care of eligible cardiac patients should be aware of the evidence for the effectiveness of cardiac rehabilitation, and researchers should examine programmatic and provider factors that affect women's participation. PMID- 12632593 TI - [Remembrance of Professor B.G. Weber (7 August 1927--22 August 2002]. PMID- 12632592 TI - Medicaid and the unconstitutional dimensions of prior authorization. PMID- 12632595 TI - Membranous nephropathy and seronegative systemic lupus erythematosus. PMID- 12632594 TI - A note on mutation analysis in familial Mediterranean fever. PMID- 12632596 TI - What recruiters won't tell you. Job seekers should limnit their expectations when working with a headhunter. PMID- 12632597 TI - Recovery of skills development levies by dentists. PMID- 12632598 TI - [Max Perutz elucidating the three dimensional structure of proteins]. PMID- 12632599 TI - Beyond the double helix. PMID- 12632600 TI - Mystery woman: the dark lady of DNA. PMID- 12632601 TI - [Digitalizaing pathways]. PMID- 12632602 TI - [XML,eXtensible Markup Language]. PMID- 12632604 TI - [Reconsideration of glucocorticoid therapy for rheumatoid arthritis]. PMID- 12632603 TI - [Heart failure to be treated according to the guidelines so that your patients live longer]. PMID- 12632605 TI - [Osteoporosis]. PMID- 12632606 TI - [Bone marrow accumulation in gallium scintigraphy in patients with adult Still's disease]. AB - We investigated the features and the usefulness of gallium scintigraphy in the diagnosis and the assessment of Adult Still's disease (ASD) by retrospective case review. Gallium scintigraphy have been done for 11 cases of ASD (3 males and 8 females) and 4 females were positive. Among these, 67 Ga-citrate was accumulated to the bone marrow in all 4 cases and to the major joints in 2 cases. Positive cases were rather serious and administered more immunosuppressants than negative cases. In order to characterize gallium scintigraphy findings of ASD, i.e. bone marrow accumulation, we analyzed 130 cases of collagen vascular diseases. Although 101 cases (77.7%) were positive, only 7 cases (5.4%) showed the accumulation of 67Ga-citrate to the bone marrow. These include 3 cases with ASD, and 1 case with systemic lupus erythematosus, polyarteritis nodosa, Wegener's granulomatosis and Sjogren's syndrome. We also accumulated 18 patients who exhibited bone marrow accumulation of 67Ga-citrate, and found that 7 patients had collagen vascular and their related diseases. In conclusion, bone marrow accumulation in gallium scintigraphy is a specific feature of collagen vascular diseases, especially ASD, and it is suggested that cases with positive gallium scintigraphy in ASD can be serious and resistant to treatment. PMID- 12632607 TI - [Use of grommet for Swanson flexible hinge toe implant arthroplasty for hallux valgus deformity of rheumatoid arthritis]. AB - OBJECTIVES: We reviewed the results of arthroplasty of the great toe with hallux valgus in rheumatoid patients, using a Swanson flexible hinge toe implant protected by grommet. MATERIALS AND METHODS: Sixteen patients (26 feet) with rheumatoid arthritis were operated on from 1996 to 1999. (fifteen women, one man). The average age was 59.8 years (range, 48-73 years). The average follow-up period was 3.0 years (range, 1.0-4.5 years). In each patient, the hallux valgus angle (HVA) was measured before and after surgery, and implant breakage and radiolucency around the implant were evaluated on radiographs. RESULTS: Average HVA was 47.7 degrees preoperatively and 19.3 degrees postoperatively. No implant breakage was observed in 76.9% of the feet. Radiolucency of more than 2 mm was observed in only 3.8% of the feet. CONCLUSIONS: These findings suggest that arthroplasty using a Swanson flexible hinge toe implant with a grommet is useful for treatment of hallux valgus in rheumatoid patients. PMID- 12632609 TI - [Intravenous cyclophosphamide pulse therapy for refractory juvenile dermatomyositis]. AB - We described three children with juvenile dermatomyositis (JDM) refractory to the conventional therapy. They were successfully treated with intravenous cyclophosphamide (IVCY) pulses, and two of them were administered plasma exchange (PE) before IVCY. Case 1. A 17-year-old girl with JDM was previously treated for 2 years with the combination of prednisolone, intravenous gamma-globulin, methotrexate, and azathioprine. However, muscle weakness gradually progressed. She failed to hold her sitting position and to rise her arms, but both serum CK and aldolase were stable. After the episode of aspiration pneumonia the follow-up muscle biopsy was performed, which revealed muscle degeneration and massive mononuclear cell infiltration in perivascular area. The erythrocyte sedimentation rate (ESR) and fibrin degradation product E (FDP-E) levels were gradually increased. Because the active inflammation of muscle and muscle vasculature was suspected, the PE and IVCY combination therapy was administered. During the 6 courses of the therapy, muscle weakness was markedly improved so that she could hold herself at the sitting position and could have meals by herself. Case 2. A 5 year-old boy with JDM was treated for 8 months with prednisolone p.o., but his muscle strength became worse. The muscle enzyme levels, such as serum CK and aldolase, were not reflecting his status of the disease, but FDP-E levels were increased. Muscle MRI and biopsy revealed the inflammatory changes of perivascular area of muscle. The PE and IVCY combination therapy was effective, and he became able to walk and run by himself. Case 3. A 14-year-old boy was diagnosed as having JDM when he was 10 years of age, and treated with prednisolone p.o., and subsequently with intravenous methylprednisolone pulses and azathioprine. Three years later the flares were observed accompanied with the elevations of serum CK and FDP-E. The administration of IVCY improved muscle strength as well as serum muscle enzyme and FDP-E levels. These findings indicated that the clinical manifestations of JDM should be closely monitored, that the serum levels of muscle enzymes including CK and aldolase were sometimes not indicative for the flares of JDM, and that muscle MRI and re-biopsy examination were needed for the children with progressive muscle weakness. In addition, determination of ESR and FDP-E was not specific but helpful to detect flares of the disease in some cases. PMID- 12632608 TI - [A possible role of anti-endothelial cell antibody in the sera of MCTD patients on pulmonary vascular damage relating to pulmonary hypertension]. AB - OBJECTIVES: Pulmonary hypertension (PH) is one of the major fatal causes in patients with mixed connective tissue disease(MCTD), which showed remarkable angiopathy from large to small vessels in the lungs. However, the etiology of PH in MCTD is still unknown. Even the lung tissues of MCTD patients without overt clinical PH represent minor vascular damages such as microthrombus or slight intimal thickening. These findings suggest some serum factors cause endothelial cell damage especially to pulmonary micro vessels, which leads to PH in MCTD. To elucidate the mechanisms of PH in MCTD we studied the anti-endothelial cell antibodies (AECA) in the sera of MCTD patients, which are considered to correlate with activity in some collagen diseases, and compared the three kinds of endothelial cells, especially the effects on pulmonary endothelial cells. MATERIALS AND METHODS: Sera from 14 MCTD patients who satisfied the Kasukawa's criteria in Japan including 4 cases of PH, 8 cases of non-PH and 3 untreated cases, and 5 healthy controls were analyzed as follows: (1) AECA to human pulmonary arterial endothelial cells (HPAEC), pulmonary microvascular endothelial cells (HMVE-L) and human aortic endothelial cells(HAEC) were analyzed by an indirect immunofluorescence method using flow cytometry. (2) Effects of MCTD patients' and healthy controls' sera on cell proliferation and induction of apoptosis on cultured HPAEC were investigated by methods of MTS and TUNEL. (3) A cytotoxic effect of patients' sera in combination with activated NK cells on HPAEC were studied by a method of LDH concentration. RESULTS: (1) Patients' sera from MCTD have IgG type AECA, and sera from MCTD patients with PH showed a higher intensity of AECA compared with non-PH and control cases (P < 0.01). (2) Only patient's sera revealed no potency of cell proliferation and induction of apoptosis in every kinds of endothelial cells compared with controls. (3) Sera from MCTD patients with PH, and from untreatment patients were high intensity of AECA, which shows cytotoxicity by addition of activated NK cells. CONCLUSIONS: Apoptosis of pulmonary arterial endothelial cells induced by AECA in combination with activated NK cells may be the fist step of vascular damage associated with pulmonary hypertension in patients with MCTD. PMID- 12632610 TI - [A case of scleroderma with pancytopenia due to lack of trace elements]. AB - A 66-year-old woman with systemic scleroderma developed pancytopenia (white blood cell 750/microliter, neutrophilic cell 201/microliter, red blood cell 166 x 10(4)/microliter, hemoglobin 5.3 g/dl, hematocrit 18.1%, platelet cell 8.2 x 10(4)/microliter) 7 months after the initiation of intravenous hyper-alimentation for chronic ileus. Serum copper and zinc levels were 3 and 46 micrograms/dl, respectively. Provision of trace elements led to increase blood cell counts as well as serum copper and zinc levels. She also developed watery diarrhea frequent times a day and hypoproteinemia during the lack of trace elements. Evidence of protein-losing gastroenteropathy was shown by gastrointestinal scintigraphy using 99mTc-human serum albumin half a year after provision of trace elements and it was not shown one and a half years after continuous provision of trace elements. As patients with scleroderma sometimes develop gastrointestinal problems and are needed intravenous nutrition of long duration, they should be paid attention to lack of trace elements that can be a cause of hematologic complications. PMID- 12632611 TI - [Systemic sclerosis associated with microscopic polyangitis presenting with high myeloperoxidase (MPO) titer and necrotizing angitis: a case report]. AB - We herein report a case of systemic sclerosis associated with microscopic polyangitis. The patient was a 54-year-old woman, who was diagnosed to have systemic sclerosis at a hospital in 1992, but she did not receive any medical treatment. She had been suffering from pyrexia, paresthesia and muscle weakness of both lower limbs since the beginning of 2001, and was introduced to our hospital. She showed hardened skin extending from her fingers to upper arms, weakness in both lower limbs and livedo reticularis. Her laboratory test showed WBC 11, 600/microliter, CRP 6.63 mg/dl, CH 50 24 U/ml, anti Scl-70 antibody 90.1 index, and MPO-ANCA 281 EU, but no impaired renal function was recognized. Chest computed tomography showed interstitial pneumonia while necrotising vasculitis of the right sural nerve was found in a biopsy specimen. Based on these findings, we diagnosed her to have systemic sclerosis accompanied with microscopic polyangitis (MPA). She received steroid treatment after the diagnosis was made, and her symptoms and the laboratory findings thereafter immediately improved. Many cases have been reported to have ANCA positive systemic sclerosis among patients with systemic sclerosis that are complicated MPO-ANCA-related vasculitis. However, since our patient demonstrated necrotising vasculitis in a sural nerve biopsy and no evidence of an impaired renal function, we diagnosed her to have systemic sclerosis complicated with MPA instead of ANCA positive systemic sclerosis. The pathological state of this patient thus seemed to be different from that of ANCA positive systemic sclerosis. We concluded that this patient had both systemic sclerosis and MPA. It is therefore important to note that some patients who have been reported to have ANCA-positive systemic sclerosis may also have systemic sclerosis complicated with MPA. PMID- 12632612 TI - [The role of glycine binding site in NMDA receptor--interactions between NMDA and D-serine in artificial anoxia/agycemia rat hippocampus]. AB - BACKGROUND: NMDA receptor contributes to cause neuronal death in anoxic condition. It is not known how a part of NMDA receptors, NMDA-binding site and/or glycine-binding site, influence neuronal damage in rats' hippocampus in vitro. METHODS: Rats' hippocampus, labeled with norepinephrine (3H-NE), was incubated in artificial cerebrospinal fluid (aCSF) and we measured 3H-NE in superfusion solution and remaining tissue. Glucose was eliminated from aCSF and 95% N2 + 5% CO2 produced the anoxic state. RESULTS: The amount of 3H-NE release increased in anoxia with NMDA (NMDA-binding site agonist), while there was no influence on NMDA receptor in non-anoxic state even after D-serine (glycine-binding site agonist) has been administered. The 3H-NE was released more when D-serine (100 mu mM) and NMDA (100 mu mM) were administered together than when only D-serine (10 mu mM, 100 mu mM, 1000 mu mM) in anoxia or NMDA (10 mu mM, 100 mu mM, 1000 mu mM) in anoxia was administered. CONCLUSION: Glycine-binding site agonist alone does not act significantly but ion channels in NMDA receptor open more and become more effective when both glycine-binding site agonist and NMDA-binding site agonist exist, suggesting that there are interactions between NMDA-binding site and glycine-binding site in NMDA-receptor during anoxia. PMID- 12632613 TI - [A successfully treated case of intraoperative latex anaphylaxis during abdominal aorta aneurysm resection]. AB - We experienced resuscitation from latex anaphylaxis induced by surgical gloves during elective abdominal aorta aneurysm resection in a low risk adult with latex allergy. The patient developed severe circulatory collapse 10 minutes after the start of surgery. Although administration of dopamine, norepinephrine, and a large amount of fluid could not normalize this circulatory collapse, but addition of famotidine and chlorpheniramine could restore blood pressure. After resuscitation, we decided to postpone the operation. The re-operation was planned in the latex free setting one month later. All products, containing latex, had been excluded from operating room one day before the re-operation, and the patient entered operating room earlier than other patients to avoid latex polluted powder from surgical gloves of other staffs. Y-shaped graft replacement was performed, and the perioperative course was uneventful. Latex is the second leading cause of intraoperative anaphylaxis, and the high-risk cases could be easily detected through the questionnaires, focusing on latex and other tropical fruits allergy. Moreover, because it is easy to get latex-free products, we recommend that a set of latex free products should be prepared at least for a high-risk case of latex allergy. PMID- 12632614 TI - [Intensive care management of acute pulmonary thromboembolism]. AB - Three patients with acute pulmonary thromboembolism, who had fallen into cardiopulmonary arrest or severe respiratory failure, were treated and saved in our intensive care unit. Two patients were resuscitated with percutaneous cardiopulmonary support device. Two patients underwent surgical embolectomy and we carefully applied positive pressure ventilation to prevent postoperative reperfusion pulmonary edema. Early diagnosis and treatment are essential for saving critically ill patients with acute pulmonary embolism. However, cardiopulmonary resuscitation is often difficult and unsuccessful. Therefore prophylaxis is strongly recommended especially in patients with known risk factors of venous thrombosis. Since reperfusion pulmonary edema is a serious complication after surgical pulmonary embolectomy, careful postoperative respiratory care is needed. PMID- 12632615 TI - [The effects of colloid preload on hemodynamics and plasma concentration of atrial natriuretic peptide during spinal anesthesia in elderly patients]. AB - We evaluated the effects of moderate colloid preloading on hemodynamics and plasma concentration of atrial natriuretic peptide (ANP) during spinal anesthesia in elderly patients undergoing low extremity surgery. Twenty patients (aged 66-90 yr) were randomly divided into two groups. Control group (n = 10) received no prehydration, and hydration group (n = 10) received colloid (6% hydroxyethyl starch; HES) preloading of 8 ml.kg-1 before spinal anesthesia. Systolic blood pressure decreased significantly 10 and 30 min after spinal anesthesia in either group, and there was no difference between the groups in the incidence of hypotension. The concentration of ANP decreased significantly by 23% in control group, whereas it increased significantly by 86% in hydration group, suggesting that cardiac preload might increase with volume expansion effect of prehydration with HES. In conclusion, colloid preloading with moderate volume might prevent the decrease in cardiac preload with increasing ANP, whereas it did not prevent spinal-induced hypotension in elderly patients. PMID- 12632616 TI - [Effect of teeth protector on dental injuries during general anesthesia]. AB - BACKGROUND: Dental injuries are the most common reason for complaints against anesthetists. The purpose of this study was to investigate the effect of teeth protector on dental injuries during general anesthesia. METHODS: Incidence of dental injuries was evaluated retrospectively in 5,946 consecutive patients between November 1998 and October 2001. All the patients were interviewed directly at a post-anesthetic clinic. RESULTS: Dental injuries were observed in 2.1% of the patients, and the injuries occurred more frequently in the patients undergoing difficult tracheal intubation. The teeth protector used in the present study was made of cellulose aceto-butylate and was produced using eruko-pressed disks molded from own teeth form. One hundred and eighty five patients (3.1%) requested teeth protectors, and none of them had their teeth injured during anesthesia. CONCLUSION: Our results showed that the teeth protector could protect the teeth from dental injuries and increased the satisfaction with anesthesia. Preoperatively anesthesiologists should know the characteristics of patients' teeth and inform the patients of the risk of injury. We conclude that the custom made protector might be useful and necessary to preserve the teeth from dental injuries during anesthesia. PMID- 12632617 TI - [Can the salvaged autologous blood transfusion be safe during TIVA?]. AB - BACKGROUND: This study aimed to determine whether the salvaged blood can be safely used after surgery and to examine the blood concentration of anesthetics in salvaged blood processed by autologous transfusion device (Cell Saver 5, Haemonetics Corp.) during propofol anesthesia. METHODS: Ten consenting patients (aged 48-79 yr, classified ASA physical status I or II) scheduled for total hip arthroplasty were randomly divided into two groups: in the first group, slowing pattern of electroencephalographic activity (moderate-dose group: M) was obtained and in the second group, a burst-suppression (high-dose group: H) was obtained. Employing the electroencephalogram (EEG) and spectral edge frequency, propofol was titrated to maintain the EEG pattern. All patients received propofol anesthesia combined with lumbar epidural anesthesia. RESULTS: The infusion rates of propofol at the end of surgery were 9.6 +/- 1.1 (H) and 7.0 +/- 1.4 mg.kg-1.h 1(M), respectively. The blood concentration of propofol in the H group was significantly greater than that in the M group. But no significant difference was observed in the salvaged blood concentration of propofol between the two groups (H: 0.391 +/- 0.158 microgram.ml-1, M: 0.401 +/- 0.236 microgram.ml-1). CONCLUSION: The concentrations of propofol in salvaged blood were low, irrespective of the different infusion rates. These results demonstrate that salvaged autologous blood would not contribute to sedative or anesthetic effect. PMID- 12632618 TI - ["Tube scratching" in the trachea on nasotracheal intubation: comparison of Macintosh and McCoy laryngoscopes]. AB - BACKGROUND: "Tube scratching" in the trachea, the intratracheal resistance of a tube, on nasotracheal intubation was evaluated using the Macintosh and the McCoy laryngoscope blade. METHODS: Fifty young patients requiring nasotracheal intubation were studied. Following induction of anesthesia, X-ray was taken before and during the use of two types of laryngoscopes. For analysis of the configuration of the upper airway, we drew three straight lines [axis of pharynx (P), larynx (L) and trachea (T)] on each film. "Tube scratching" was evaluated and graded to four by a supporting anesthesiologists. RESULTS: "Tube scratching" in the trachea was observed in 14/25 patients with McCoy laryngoscope and 4/25 patients with Macintosh laryngoscope (P < 0.05). Intubation with McCoy laryngoscope decreased more the angles formed by P and L, and L and P those that with Macintosh laryngoscope (P < 0.01). These data indicate that the lines of P, L and T tend not to align with McCoy laryngoscope, preventing the tracheal tube to be passed smoothly. CONCLUSION: An excessive lifting of the epiglottis, often observed in using McCoy laryngoscope, causes "Tube scratching" in the trachea on nasotracheal intubation. PMID- 12632619 TI - [Anesthetic management of two patients with hypertrophic obstructive cardiomyopathy for percutaneous transluminal septal myocardial ablation]. AB - We experienced anesthetic management of two patients with hypertrophic obstructive cardiomyopathy (HOCM) for percutaneous transluminal septal myocardial ablation (PTSMA). PTSMA had been performed more than a year earlier in both cases, and symptoms and exercise tolerance were improved after PTSMA. Laparotomy for suspected ovarian cancer in one patient and thoracotomy for metastatic lung cancer in the other patient were proposed. Preoperative echocardiography showed reduced left ventricular outflow tract pressure gradient (from 90 mmHg before PTSMA to 10 mmHg and from 81 mmHg to 17 mmHg, respectively) and decreased septal wall thickness. Systolic anterior movement of mitral valve apparatus had disappeared. General anesthesia in the former and general anesthesia combined with epidural anesthesia in the latter were employed. Anesthesia was induced with propofol and fentanyl. A laryngeal mask was used to minimize hemodynamic fluctuations during induction and emergence in the former. Anesthesia was maintained with propofol, sevoflurane and supplemental fentanyl. Epidural anesthesia was also used to maintain anesthesia in the latter. The postoperative course was uneventful in each case. PTSMA for HOCM might be useful to prevent perioperative cardiac events. PMID- 12632620 TI - [Hypotension after turning to the prone position]. AB - Hypotension after positioning is sometimes seen especially in patients with cervical spinal lesion operated on under prone position. Patients with spinal lesion and those with brain lesion are compared in the frequency of hypotension after positioning to prone. Sixty-one cases operated on with prone position were studied. Ages ranged from 40 to 82 (mean 61) years and ASA grade was 1 or 2 in each case. Cervical laminoplasty (group C) or craniotomy (group B) are performed in 40 and 21 patients, respectively. Ephedrine was administrated when the systolic blood pressure decreased under 80 mmHg and the frequency of ephedrine use was compared. There were no differences in age and sex distribution between group C and B. The induction doses of propofol and fentanyl in group B were larger than those of group C, but ephedrine use in group C was more frequent than in group B. In T2-weighted image of the cervical cord, high signal intensity areas were depicted in cases with hypotension. The sympathetic flow descends in the medial part in the lateral funiculus. Damage of this pathway would cause autonomic dysfunction in patients with cervical spinal lesion and strict monitoring is necessary during positioning to prone. PMID- 12632621 TI - [Anesthetic management of a patient with lymphangioleiomyomatosis who developed respiratory failure post-operatively]. AB - A 34-year old female underwent lung biopsy under video assisted thoracoscopic surgery for lymphangioleiomyomatosis (LAM). She had obstructive lung disease, and had a large amount of ascites. We did not treat her lung disease pre-operatively because her pulmonary symptom was not severe. During operation, anesthesia was uneventful even during one lung ventilation period. After surgery, she showed hypoxemia and fell into respiratory failure. We suspect that respiratory failure was induced by ascites in this case. Respiratory failure would have come from restrictive ventilatory impairment caused by a large amount of ascites in addition to the obstructive ventilatory impairment. Care should be taken on respiratory function in case of LAM with ascites during perioperative period. PMID- 12632622 TI - [Recruitment maneuver and high PEEP in a patient with severe reperfusion pulmonary edema after pulmonary thromboendarterectomy]. AB - Recently open lung approach such as recruitment maneuver and high PEEP has been applied in patients with acute respiratory distress syndrome. We report here a 29 year-old man with chronic pulmonary thromboembolism (CPTE). In this case, recruitment maneuver and high PEEP relieved postoperative respiratory failure. The major complications after pulmonary thromboendarterectomy (PTE) for CPTE are: reperfusion pulmonary edema (RPE), pulmonary hypertension (PH), hypoxia and bronchial bleeding, all of which the patient has demonstrated. Preoperative examination showed high pulmonary artery pressure (85/41, mean 50 mmHg), and high pulmonary vascular resistance predicted poor postoperative course. After PTE, he developed severe RPE, PH, hypoxia and bronchial bleeding, resulting in failed weaning from cardiopulmonary bypass. Therefore he required mechanical support of percutaneous cardiopulmonary support (PCPS). In ICU we repeated recruitment maneuver (PEEP: 30 cmH2O, peak inspiratory pressure: 42 cmH2O, respiratory rate; 15 breaths.min-1, for 1 min) and kept high PEEP (15 cmH2O). After initiating this ventilatory strategy, RPE was gradually relieved, followed by improvement of oxygenation and PH. We successfully weaned him from PCPS 38 hr after surgery and he was discharged alive on the 90 post-operative day. We conclude that recruitment maneuver and high PEEP may be useful for RPE developed after PTE. PMID- 12632623 TI - [Septic shock associated with pyogenic liver abscess rescued with percutaneous transhepatic abscess drainage]. AB - We report a case of septic shock associated with pyogenic liver abscess rescued with percutaneous transhepatic abscess drainage (PTAD). A 70-year-old male patient was admitted to our outpatient department of internal medicine with general fatigue, dullness of bilateral shoulders and extremities, appetite loss, weight loss, headache, and vertigo. Laboratory tests showed severe inflammatory indications, anemia, and high values of hepatobiliary enzymes and blood sugar. Abdominal ultrasonography and enhanced CT showed a pyogenic liver abscess of 10 cm in diameter at S 6-7 in the right hepatic lobe. The patient's condition deteriorated suddenly that night. From the results of abdominal ultrasonography and enhanced CT, we made diagnosis of septic shock associated with pyogenic liver abscess. Emergency abdominal ultrasound-guided PTAD was performed under local anesthetic. Postoperatively, the antibiotic was infused daily through a PTAD tube into the liver abscess space. He recovered and his laboratory tests improved gradually. On abdominal ultrasonography and enhanced CT, the liver abscess disappeared by 19th postoperative day, and PTAD tube was removed. There was no complication during PTAD treatment. We conclude that patients in septic shock should undergo further examinations immediately and treatment of the infected tissue should be started as soon as possible. PTAD may be an additional effective procedure for pyogenic liver abscess in septic shock. Furthermore, local antibiotic lavage through a PTAD tube into the liver abscess space may be an important supplementary method in the management of the illness. PMID- 12632624 TI - [A case of pulmonary embolism and a case of ileus as complications after laparoscopic radical prostatectomy]. AB - In recent years, the use of laparoscopic techniques for surgical operations has been increasing, because this procedure is less invasive and is excellent in regard to patient's quality of life. Normally, complications are rare in laparoscopic surgery. However, we experienced a case of pulmonary embolism and one case of ileus as complications after laparoscopic radical prostatectomy. Especially, in this type of operation, the danger of complications is increased due to the severe head down and lithotomy position, which is employed to ensure a good view during operation. In this particular case, the long duration of operation may have been another related risk factor. There were no risk factors for pulmonary embolism such as those encountered when a patient is aged, obese, or bed ridden for a long time. However, an intermittent air massage must be applied to the lower legs to prevent thrombus due to poor blood circulation of the lower extremities below the knee during the surgery. It is also necessary to change the posture of the patient frequently after the operation. In addition, the administration of low molecular weight heparin may also be effective. PMID- 12632625 TI - [Propofol anesthesia for a patient in the terminal stage of Fukuyama type congenital muscular dystrophy]. AB - A 31-year-old tracheostomized woman weighing 32.5 kg with Fukuyama type congenital muscular dystrophy in terminal stage was scheduled for dilatation of the tracheostoma, removal of the tracheal granulomas and reconstruction of gastrostoma. Anesthesia was induced and maintained with continuous propofol infusion and intermittent fentanyl. Muscle relaxant was not necessary because of generalized severe muscular atrophy. There was no hemodynamic derangement during the surgery and emergence from the anesthesia was rapid. There were no postoperative complications related to anesthesia and surgery. PMID- 12632626 TI - [Severe hypotension and atrio-ventricular block in a patient of left lung cancer associated with myocardial bridging]. AB - A 74-year-old man with myocardial bridging was referred to our hospital for operation of the left lung cancer. He underwent upper lobectomy of the lung under general anesthesia. After lobectomy and bilateral lymph node resection, severe hypotension occurred without ECG change. The blood pressure was restored by cardiac massage and the administration of fluids and vasoactive agents. After the closure of the sternum, hypotension occurred again and complete A-V block appeared. After resuscitation, A-V block disappeared. He was extubated the day after surgery without any neurogical deficit. We consider that hypovolemia and myocardial bridging induced hypotension and complete A-V block. PMID- 12632627 TI - [Anesthetic management of a patient with congenital plasminogen activator inhibitor-1 deficiency]. AB - Congenital plasminogen activator inhibitor-1 (PAI-1) deficiency is an extremely rare disorder characterized by a bleeding diathesis due to hyperfibrinolysis as a result of decreased PAI-1 activity. A 21-year-old male with congenital PAI-1 deficiency underwent wisdom teeth extraction of the mandible under general anesthesia using propofol, nitrous oxide, sevoflurane, fentanyl, and vecuronium. No complications including prolonged bleeding and rebleeding after the operation were observed because hemostatic management was successful by using intravenous tranexamic acid. PMID- 12632628 TI - [Food residues identified in the pharynx of a patient with a cleft palate]. AB - Solid food residues were identified in the pharynx of a 17-month-old infant with a cleft palate at the time of anesthesia induction, although nothing-by-mouth directions for more than four hours had been strictly followed. Preoperative airway management, e.g. gargling and nose blowing, is necessary in patients with a cleft palate even after an appropriate period of fasting because food residues may remain in the nose due to the anatomical abnormality. PMID- 12632629 TI - [A novel whole blood aggregometer]. AB - We examined ADP-induced platelet aggregability in 45 patients by novel whole blood aggregometer based on the screen filtration pressure method. Platelet aggregation could be easily and immediately checked without preparing platelet rich plasma. This aggregometer was useful for determination of anesthetic methods for a patient with an anti-platelet therapy. However, PATI (platelet aggregately threshold index) demonstrated the lowest value 5 minutes after blood collection and the valve was stabilized at higher level for 30 to 120 minutes. Time dependent change should be considered for assessment of platelet aggregability in a clinical study. PMID- 12632630 TI - [Evaluation of multi-modality treatment for the patients with advanced esophageal cancer]. AB - Hyperthermia has been proved to be intensely cytotoxic to malignant cells when combined with anticancer agents and irradiation. We have applied hyperthermo chemo-radio-therapy (HCR) using a radiofrequency system with an endotract electrode to the patients with advanced esophageal cancer. In this study, 132 patients with advanced esophageal cancer with invasion to the neighboring structures, who underwent non-curative operation, were analysed. The markedly effective cases was observed in 12.2% in HCR group, while in 2.5% in chemoradiotherapy (CR) group, microscopically. The survival rate of the HCR group was significantly better than that of the CR group (p < 0.05). Pre- and post operative leukocytopenia caused by preoperative treatment was not found to be a risk factor of postoperative complication. It is important to continue to clarify the factors influencing the effectiveness of preoperative HCR in the future. We hope that HCR will play an even more important role in treatment of esophageal cancer. PMID- 12632631 TI - [Living donor liver transplantation for a patient with renal failure]. AB - Pretransplant renal failure is a well-known risk factor which adversely affects the prognosis after liver transplantation. We report a case with pretransplant renal failure and discuss the perioperative management of such patient. The patient was 62 year-old-woman who was diagnosed with end-stage liver disease due to primary biliary cirrhosis, for which living donor liver transplantation (LDLT) was indicated. Her pretransplant serum creatinine was 9.4 mg/dl due to combination of drug-induced (antibiotics) acute tubular necrosis and hepatorenal syndrome. The management of renal failure consisted of the avoidance of calcineurin inhibitor as an induction immunosuppression and the use of perioperative continuous hemodiafiltration (CHDF). The postoperative course of the patient was complicated with CMV pneumonia and acute rejection, however she recovered and discharged on 94 POD with well-preserved graft function and normal renal function without any adverse sequela. LDLT for patients with renal failure can be performed successfully by careful management. PMID- 12632633 TI - Forensic pathology and the Caribbean. PMID- 12632632 TI - Leprosy elimination campaigns: impact on case detection. PMID- 12632634 TI - Seeking a commitment for action: AIDS--2002. PMID- 12632636 TI - Chronic renal failure in Jamaican children. AB - In order to document the incidence, aetiology and outcome of chronic renal failure in Jamaican children, paediatric surgeons and hospital based paediatricians island-wide were contacted, and the nephrology records at the island's paediatric nephrology centres searched for data on children < 12-year old with chronic renal failure diagnosed for the first time between January 1985 and December 2000. Thirty-four children were identified, 21 were male. The cumulative annual incidence of chronic renal failure was 3.2 per million children aged < 12 years. The incidence is likely underestimated, as some children may have been undiagnosed and/or not referred. Glomerulonephritis was the commonest cause of chronic renal failure (50%) followed by obstructive uropathy, reflux nephropathy, renal dysplasia and chronic pyelonephritis (41.2%). Five children (14.7%) had reflux nephropathy (post obstructive in four). Half the children were already in chronic renal failure at time of presentation. Mortality was 65%. In Jamaica, childhood chronic renal failure is due mainly to potentially treatable diseases. Local physicians should be more aware of potentially progressive renal diseases and their prevention. Earlier referral of difficult cases for nephrological consultation is recommended. A paediatric dialysis/transplant programme is needed. PMID- 12632635 TI - Evidence of collateralization of vagal efferents innervating subdiaphragmatic segments of the gastrointestinal tract in the rat using the double labelling fluorescent dyes technique. AB - Collateralization of the abdominal vagal trunks was investigated in the rat using double labelling fluorescence dye technique. A total of 20 adult male and female rats were used for the study. The anterior and posterior walls of the corpus and fundus of the stomach were injected with 0.1 microliter (microliter) of 5% aqueous solution of diamidino yellow (Dy) in eight rats. The same quantity and percentage of fast blue (Fb) was injected into the walls of the duodenum and proximal jejunum in the same eight rats that were injected with Dy. The anterior and posterior walls of the stomach of four rats were injected with 0.1 microliter of 5% Dy only while four other rats had injections of 0.1 microliter of Fb only into the duodenum and proximal jejunum. Two control rats had stomach injections with normal saline, and two rats had saline injections into the intestine. Each rat was perfused with a preservative 14 days after injection and serial sections taken for examination with a fluorescence microscope. The results of the experiment revealed that in the eight rats injected simultaneously with Dy and Fb, some neurons of the dorsal motor nucleus of the vagus nerve (DMX) were labelled with Dy only, some with Fb only and some were doubly labelled with Dy and Fb. No double-labelled neurons were seen in the rat injected with one dye only and no labellings at all were seen in the controls. The pattern of labelling obtained in the study is suggestive of collateralization of axons of the abdominal vagal trunks. PMID- 12632637 TI - Value of routine long bone radiographs in management of babies with a positive VDRL at the Mount Hope Women's Hospital. AB - This retrospective study examined the records of 44 mothers and 44 babies with a positive blood VDRL test and/or who showed symptoms of congenital syphilis at the Mount Hope Women's Hospital (MHWH), Trinidad, between June 1, 1996 and September 30, 1998. Long bone radiographs of 40/44 (91%) babies were assessed independently by two radiologists for signs of congenital syphilis and the data were analysed using frequency tables. Of the 44 mothers in the study, 34 (77%) were VDRL positive and the rest were either negative or unknown. Maternal age ranged from 14 to 39 years and most (92%) had antenatal care. Fourteen of the 34 (41%) VDRL positive mothers received treatment antenatally. All mothers were asymptomatic and none had any clinical features of syphilis. In 35/44 babies, the VDRL test was positive and in nine, the test was either negative or unknown. Twenty-six of the babies had signs suggestive of congenital syphilis and in 12, (46%) long bone radiographs revealed changes compatible with the diagnosis of congenital syphilis. Radiographic abnormalities were present only in infants with VDRL titres > 1:8. PMID- 12632638 TI - Smooth muscle tumours of the vulva in Jamaica. AB - Smooth muscle tumours of the vulva are uncommon and their behaviour is unpredictable. A 30-year retrospective analysis of the pathology files and patient records revealed 10 cases diagnosed at the University Hospital of the West Indies, Kingston, Jamaica, with one malignant and one recurrent tumour. Size, infiltrating margins and mitotic count are important parameters to determine prognosis. Although these tumours are of low malignancy, mitotic count may be mandatory in determining their potential for recurrence. PMID- 12632639 TI - Histopathological findings in women with postmenopausal bleeding in Jamaica. AB - We investigated the histopathological findings in women presenting with postmenopausal bleeding in a population predominantly of African descent by conducting a six-year retrospective study of 716 gynaecological surgical specimens from 629 women accessed in the Department of Pathology, The University of the West Indies, Jamaica. Histopathological diagnoses were correlated with patients age, specimen volume, duration of bleeding and length of postmenopausal interval at presentation using t-tests and linear regression models. The mean (SD) age was 63.6 (9.3) years. The frequency of the main causes of postmenopausal bleeding was: endometrial hyperplasia (22.3%); endometrial atrophy (21.3%); non diagnostic (19.9%); endometrial carcinoma (9.5%); cervical carcinoma (6.8%); cervical polyps (4.5%); endometrial sarcoma (3.5%); proliferative endometrium (3.2%). Mean (SD) duration of bleeding was 6.9 (12.7) months. The percentage of women with malignant lesions was two to three times greater than those reported in the United States of America and Europe subsequent to 1980. The delay between the onset of bleeding and presentation is worrisome and suggests the need for public education. PMID- 12632640 TI - Compliance and blood pressure control in women with hypertension. AB - Compliance with treatment is a fundamental prerequisite for therapeutic benefit. The aim of this study is to determine the level of knowledge of hypertension, compliance with recommended antihypertensive therapy, and current blood pressure status in women with hypertension attending a Type V health centre. A pre-tested questionnaire with 37 in-depth items was administered to 30 (37.5%) women, selected by quota sampling, from a population of 80 women with hypertension, on four consecutive regular clinic days in May/June 2001. Weights and the mean of two blood pressure measurements were recorded. Data were analysed using Statistical Package for the Social Sciences (SPSS) version 7.5. The median age and weight of the respondents was 57 years (range 36-85 years) and 80.3 kg (range 66.8-150 kg). Median duration of hypertension was five years. Fifty per cent of the sample were diabetic. The longer the patient had been hypertensive, the greater the compliance with medication (p < 0.05). Twenty per cent of non diabetics were controlled to blood pressure < or = 140/90 mmHg and 13% of the diabetics were controlled to blood pressure < or = 135/85 mmHg. Twenty per cent reported ill effects from medication; 60% used "folk remedies" such as garlic. Only 27% of patients were fully compliant with medication. Sixty per cent did no exercise, 73% did less than one hour of exercise per week. Diabetics took more exercise than non-diabetics (Z = -2.1, p < 0.05) and were more compliant with medication than non-diabetics (Z = -2.3, p < 0.05). All respondents included salt in their diets and consumed fruits and vegetables only "sometimes". One third believed that hypertension could be "cured". The overall median knowledge score and median compliance score were 50% (range 16.7%-100%) and 31% (range 13%-60%) respectively. This group had inadequate knowledge of hypertension, poor compliance with recommended antihypertensive therapy (JNCVI) and limited BP control. Counselling of these patients in the areas of medication, diet, exercise and weight control is recommended. Further research, using randomized samples, to inform interventions to improve the knowledge, compliance and self-care management of patients with hypertension is indicated. PMID- 12632641 TI - Preoperative starvation and pulmonary aspiration. New perspectives and guidelines. AB - The fear of aspiration of gastric contents and its life-threatening consequences in patients(aspiration pneumonitis and respiratory failure), has caused many medical practitioners, particularly anaesthetists, to rigidly follow conservative (i.e. prolonged) preoperative fasting standards. This is the nil per os (NPO) order for clear fluids/liquids and solids overnight or six to eight hours preceding the induction of anaesthesia. This practice neither takes into account the differences in the rate of gastric emptying for solid food (which may exceed six hours) and clear liquids (which is one to two hours), nor the differences in scheduled times of surgery. Long-term prospective studies and retrospective reviews have shown that the incidence of significant clinical aspiration is low: 1.4-6.0 per 100,00 anaesthetics for elective general surgery. Risk factors for pulmonary aspiration include: a high American Society of Anaesthesiologists (ASA) physical status score; emergency surgery; difficult airway management; increased gastric volume and acidity; increased intra-abdominal pressure; gastro oesophageal reflux; oesophageal disease; head injury with impaired consciousness and extremes of age. Experimental studies and reviews have consistently shown the safety of clear liquid ingestion up to two hours before induction of anaesthesia in healthy patients without risk factors, and the fact that a longer fluid fast does not necessarily offer any added protection against pulmonary aspiration. The conservative pre-operative fasting standard causes discomfort and in some cases, suffering of patients and is therefore unnecessary for patients without risk factor(s). Anecdotal reports at the University Hospital of the West Indies (UHWI) have shown that application of the liberalized guidelines for preoperative fasting and fluid intake has not resulted in increased pulmonary aspiration, morbidity or mortality. Instead it has resulted in decreased irritability, anxiety, thirst and hunger in the peri-operative period. Patients, especially children are more comfortable and the perioperative period is better tolerated. It is therefore time that all medical personnel adopt the liberalized guidelines. PMID- 12632642 TI - Image and diagnosis. Cutaneous calcinosis (calcinosis cutis). PMID- 12632643 TI - Progressive Epstein-Barr virus infection in a six-year-old male. AB - Progressive Epstein-Barr virus infection is described in a six-year-old Jamaican male with no previous evidence of immune deficiency. PMID- 12632644 TI - Multiple endocrine neoplasia type 2B. A case report and review of the syndrome. AB - This paper presents a case of the very rare multiple endocrine neoplasia Type 2B syndrome. It highlights that because of the presence of superficial neuromas in this condition, there is the possibility for early diagnosis. Recent knowledge of the molecular genetics of this syndrome and the ability to screen family members is also stressed since early thyroidectomy is now recommended to prevent the development of thyroid carcinoma which is the main determinant of prognosis. PMID- 12632645 TI - Angiomatosis of the breast in a male child. A case report with immunohistochemical analysis. AB - Angiomatosis is a benign vascular lesion that has been described rarely in the breast. We describe a case in a seven-year-old boy of African descent who presented with progressively increasing, unilateral breast enlargement, the first such report in a male child. The patient underwent excisional biopsy of the breast mass followed by mastectomy. Pathologic examination revealed a diffuse proliferation of variably-sized, thin-walled vascular channels lined by flattened endothelium that showed negative immunohistochemical staining for von Willebrand factor, factor VIII-related antigen, CD34 and S-100 protein. There is no evidence of recurrence after 24 months of follow-up. PMID- 12632646 TI - Cervical pregnancy. Case report and literature review. AB - Cervical pregnancy is an uncommon life threatening form of ectopic pregnancy that is associated with the unexpected occurrence of uncontrollable haemorrhage from the cervix. To avoid hysterectomy and to maintain fertility, several conservative methods of termination have been used. We present herein, one such case which relates to a young woman who had successful conservative management of a cervical pregnancy using vasopressin infiltration of the cervix, a McDonald cerclage and cervical curettage followed by balloon tamponade. Subsequently, she became pregnant again with a normal intra-uterine pregnancy and had a successful vaginal delivery. PMID- 12632647 TI - Successful pregnancy after hysteroscopic metroplasty. AB - Congenital anomalies of the uterus have been estimated to affect 0.06-10% of women. Uterine septae account for approximately 80% to 90% of these anomalies and is also associated with the poorest reproductive outcome. Pregnancy loss in patients with a septate uterus has been reported to be as high as 90% after other causes for miscarriages have been excluded. We present here the first experience in Jamaica, of a successful pregnancy outcome after hysteroscopic metroplasty in a woman with a septate uterus and three previous miscarriages. PMID- 12632648 TI - The Ilizarov technique in the treatment of tibial bone defects. Case reports and review of the literature. AB - Autogenous cancellous bone grafting has long been the hallmark of skeletal defect management. Unfortunately, the small number of donor sites in the human body constitutes an absolute limit on the quantity of fresh autogenous cancellous bone available for filling a segmental defect. In addition, the donor sites are always a source of discomfort and morbidity for the patients. Intercalary defects resulting from trauma, infection or tumour can be treated with transport of a segment of bone within the limb using the Ilizarov technique. We report on three cases of local bone transportation for intercalary tibial defects by the Ilizarov method. PMID- 12632649 TI - Chondro-epiphyseal separation of the distal humerus in the newborn. A case report and review of the literature. AB - Chondro-epiphyseal separation of the distal humerus is a rare injury, and when it occurs in the newborn, it may be difficult to diagnose and is easily mistaken for a dislocation of the elbow. The unimpressive clinical appearance of such an injury of the elbow in an infant, as well as the absence of ossific nuclei of the distal humerus in the newborn, are responsible for the dilemma in making the diagnosis. Ultrasonography, a readily available, non-invasive technique, can be used to evaluate the non-ossified epiphysis about the elbow of infants to demonstrate dislocations, fractures, and physeal separations. Closed reduction with or without percutaneous Kirschner wire fixation is the treatment of choice for these injuries. In this article, we report on a case of complete epiphyseal separation in a neonate and discuss the problems arising in its diagnosis. PMID- 12632650 TI - The University of the West Indies Medical Students' Association full members of the International Federation of Medical Students Associations a first for a Medical Student Association in the English-speaking Caribbean. PMID- 12632651 TI - Trends in prostate pathology at the University Hospital of the West Indies, Jamaica, over the last 10 years. PMID- 12632652 TI - [In vitro transfer of the aac(6')-aph(2") gene that confers aminoglycoside antibiotic resistance from Staphylococcus aureus to Enterococcus faecalis and Enterococcus avium]. AB - In nine strains of Enterococcus, frequency of resistance gene transfer from MRSA caring gene acc(6')-aph(2") was investigated. Transfer was inducted in broth at 44 degrees C for 24 h. Presence of acc(6')-aph(2") gene was confirmed by PCR method. Frequency of gene transfer was the highest for strains Enterococcus faecalis isolated from humans (0.13), significant lower for Enterococcus avium (0.007) and the lowest for strains of Enterococcus faecalis isolated from animals (0.004). PMID- 12632653 TI - [Detection of smr and qacA genes in Staphylococcus aureus strains using PCR]. AB - The 31 strains of Staphylococcus aureus were examined for the presence of smr and qacA determinants. The smr gene was found in 15 strains. Fourteen of them were MRSA resistant to quaternary ammonium compounds, ethidium bromide, and acriflavine. One was MSSA strain resistant to ethidium bromide and acriflavine. The qacA gene was found in two MRSA strains resistant to quaternary ammonium compounds, ethidium bromide, chlorhexidine and acriflavine. One of these two strains possessed both smr and qacA genes. PMID- 12632654 TI - [Resistance of Staphylococcus aureus strains to quaternary ammonium compounds and chlorhexidine]. AB - The level of susceptibility of 90 different Staphylococcus aureus strains to chosen quaternary ammonium compounds: cetyltrimethyl ammonium bromide, benzalkonium chloride and benzethonium chloride as well as to chlorhexidine digluconate were examined. The examined strains consist of three groups: hospital originated MRSA, hospital originated MSSA and non-hospital MSSA. The significant differences between these groups were observed in they susceptibility to the investigated disinfectants. The obtained MIC values showed that the most resistant were hospital MRSA strains, where 55% was estimated as resistant to cetyltrimethyl ammonium bromide, 72% were resistant to benzalkonium chloride and benzethonium chloride and 7% were resistant to chlorhexidine digluconate. Among hospital originated MSSA 3% of strains were resistant to cetyltrimethyl ammonium bromide and 6% were resistant to benzalkonium chloride and benzethonium chloride. 14% non-hospital S. aureus strains were resistant to benzalkonium chloride and benzethonium chloride. None were resistant to chlorhexidine digluconate or cetyltrimethyl ammonium bromide. PMID- 12632655 TI - [Microbiological External Quality Assurance Program (MEQAO)--evaluation of results obtained in 2001 of sanitary-epidemiological station laboratories]. AB - The aim of this study was to evaluate reliability of identification and determination of sensitivity to antibiotics and chemotherapeutics of some Enterobacteriaceae strains in 49 sanitary-epidemiological stations. All laboratories engaged in this study received 4 strains each (S. marcescens phenotype ESBL+, K. pneumoniae phenotype ESBL+, S. Typhimurium and S. Newport) previously identified in the Department of Bacteriology of National Institute of Hygiene. The laboratories included in the survey were asked to determine the genus (serovar) of each control strain using the lowest number of biochemical test recommended by Institute and to determine of the sensitivity of the identified strains to antibiotics in accordance with the guidelines of the NCCLS and domestic recommendations. Only in 2 participating laboratories the identification of one of studied strains (K. pneumoniae) was completely bad and in 9 laboratories the serological identification of S. Newport was incomplete or serological type was not correctly recognised. In the determination of the sensitivity of the control strains to chemotherapeutic agents abnormalities were found in the technique of antibiogram and test for ESBL performing and incorrect selection of disc for antibiograms as well as erroneous interpretation of the results. PMID- 12632656 TI - [Biochemical properties of enterotoxic and enterohemorrhagic E-coli strains]. AB - The purpose of this paper has been to describe the biological characteristics of thirty strains of E. coli. The E. coli strains were isolated from cases of colibacillosis in animals and from human faeces and cow milk samples. Of the thirty analyzed strains, 19 strains (63%) were found to belong to 7 serogroups: O8, O101, O138, O141, O147, O149 and O157. In 17 strains (57%) fimbriae F4 was discovered and in 1 strain (3%) the presence of fimbriae F5 and F41 was detected. Serological and biochemical researches, based on the analysis of 35 enzymatic reactions, confirmed that all strains, used in this study, belonged to the species E. coli. The strains demonstrated differences in biochemical activity for 12 substrates. It was found that strains of serotype O157: H7 had biochemical homogeneity, except in their rate of sucrose fermentation and their ability to hydrolyze arginine and sorbitol after longer incubation. On the basis of the biochemical activity, O157: H7 strains were affiliated with biotype C. In identifying serotype O157: H7, SMAC medium with sorbitol and liquid and solid media with MUG reagent were very useful. PMID- 12632658 TI - [Identification and susceptibility to antimicrobial agents of strictly anaerobic bacteria isolated from hospitalized patients]. AB - The aim of this study was to identify anaerobic strains isolated in 2001 from clinical specimens obtained from patients of Warsaw hospital and to evaluate a susceptibility of these strains to antimicrobial agents. In 2001 two hundred and twenty five clinical strains of obligate anaerobes were cultured, which were identified in the automatic ATB system (bioMerieux, France) using biochemical tests API 20 A. Drug-susceptibility of strains was determined also in ATB system with the use of ATB ANA strips. C. difficile strains were isolated on selective CCCA medium. Toxins A/B of C. difficile directly in stool specimens were detected by means of ELISA test (TechLab, USA). Fifty four strains of Gram-negative anaerobes (B. fragilis strains dominated) and 171 strains of Gram-positive anaerobes (the greatest number of strains belonged to genus Peptostreptococcus) were cultured from clinical specimens. In the cases of antibiotic-associated diarrhea 28 C. difficile strains were isolated and C. difficile toxins A/B were detected in 39 stool samples. The most active in vitro antimicrobials against Gram-negative anaerobes were metronidazole, imipenem, ticarcillin combined with clavulanic acid and piperacillin with tazobactam. Gram-positive, clinical strains of anaerobes were the most susceptible in vitro to beta-lactam antibiotics combined with beta-lactamase inhibitors (amoxicillin/clavulanate, piperacillin/tazobactam, ticarcillin/clavulanate) and imipenem. PMID- 12632657 TI - [Use of new methods for detection of beta-lactamases with extended-spectrum substrates (ESBLs)]. AB - The aim of performed examinations was to compare results of two methods applied for the detection of extended-spectrum beta-lactamases (ESBLs). Two hundred and sixty strains of Gram-negative rods were cultured from clinical specimens obtained from hospitalized patients. These strains were identified as ESBL positive on the basis of the double-disc method (DDST) according to Jarlier et al. (1988) results. Identification of strains was performed in automatic ATB Expression system (bioMerieux, France). All strains were determined using a novel test for detection of ESBL-type enzymes (DD) according to Appleton (1999). Two discs are applied in this test: with cefpodoxime (CPD) and with cefpodoxime/clavulanate (CD 01, diagnostic disc). Consistent results of two methods (DDST and DD) were obtained in the case of 166 from among 260 of examined strains (60.4%). Consistent results concerned 161 out of 222 examined strains of enteric rods (72.5%) and only 5 from among 38 of other strains (mostly belonging to the group of non-fermenting rods). On the basis of performed investigations it can be stated that the novel method of extended-spectrum beta-lactamases (ESBLs) detection (DD) according to Appleton (1999) is more objective and easier for interpretation than the double-disc synergy test (DDST) according to Jarlier et al. (DDST), which is widely applied in the routine microbiological diagnostics. PMID- 12632659 TI - [Stimulation of adhesion molecule expression in human vascular endothelium by Bacteroides fragilis toxins--influence of polymyxin B]. AB - The aim of this study was to examine the influence of polymyxin B on the level of expression of adhesion molecules E-selectin, ICAM-1, and VCAM-1 on human vascular endothelium activated with B. fragilis endotoxins or enterotoxin. Lipopolysaccharides were extracted by phenol-water method from one nonenterotoxigenic (NTBF) and three enterotoxigenic (ETBF) B. fragilis strains. LPS preparations were purified with nucleolytic enzymes and ultracentrifugation. Enteotoxin (BFT) was prepared from the supernatant of reference B. fragilis ATCC 43858 culture by precipitation with ammonium sulphate. BFT preparations were purified with the application of ion-exchange chromatography and hydrophobic chromatography. Adhesion molecule expression on the surface of human vascular endothelial cells (HMEC-1 cell line) was determined after simultaneous stimulation with bacterial compounds at the concentration of 10 micrograms/ml and polymyxin B at the concentration of 20 micrograms/ml. Endothelial cells were activated for 4 hours (E-selectin expression) or for 24 hours (ICAM-1 and VCAM-1 expression). Adhesion molecules were detected in immunoenzymatic test (ELISA) with the use of mouse, monoclonal antibodies against human ICAM-1, VCAM-1, and E selectin. The results of performed experiments suggest, that polymyxin B changes the level of adhesion molecule expression on human vascular endothelium. This antibiotic causes changes in the expression of endothelial ICAM-1, VCAM-1, and E selectin during simultaneous stimulation of endothelium with B. fragilis endotoxins or enterotoxin. In the majority of cases the addition of polymyxin B leads to the up-regulation of examined adhesion molecules. PMID- 12632660 TI - [Susceptibility to antimicrobial drugs of enterotoxin producing strains of Bacteroides fragilis isolated from different countries]. AB - Twenty two Bacteroides fragilis strains isolated from clinical samples in different countries (England, France, the Netherlands, Poland and USA) were used in the experiments. In all strains the presence of enterotoxin (fragilysin) gene was found by PCR with primers 404/407. Drug susceptibility of B. fragilis strains was determined with Etest (MICs for penicillin G, ceftriaxone, amoxicillin/clavulanic acid, imipenem, clindamycin and metronidazole). MICs were estimated in accordance to the NCCLS recommendations (1997). All tested strains were susceptible to imipenem and metronidazole. Twenty one strains were susceptible and one was intermediate susceptible to amoxicillin/clavulanic acid. Fourteen strains were resistant to ceftriaxone and five were found highly resistant to clindamycin. All examined strains were resistant to penicillin G. Four tested strains were simultaneously resistant to penicillin G, ceftriaxone and clindamycin (three French human strains isolated from postoperative wound, peritoneal fluid and bone inflammation, and one strain isolated from a pig). PMID- 12632661 TI - [Bacterial flora from bronchoalveolar lavage and occurrence of class IgG and IgA antibodies to Chlamydia pneumoniae in patients with chronic obstructive pulmonary disease (COPD)]. AB - Bronchoalveolar lavage taken from 46 patients (ranging in age from 21 to 71 years, mean 50.6 +/- 13.9) was examined for aerobic and anaerobic bacterial flora. Sera taken from 39 of patients as well as sera taken from 25 healthy blood donors of similar age (P = 0.99) were examined to determine IgG and IgA antibodies to C. pneumoniae. Bacterial flora was routinely cultured and determined using ATB computer system (bioMerieux,). IgG and IgA antibodies were tested by the enzyme immunoassays (Labsystems, Finland, Helsinki). Sera containing anti -C. pneumoniae IgG antibodies with titers of 45 EIU or higher and IgA with titers of 12 EIU or higher were considered positive. 143 of aerobic and 74 of anaerobic bacterial strains were cultured. Streptococci group viridans, pneumococci, enteric bacilli, Haemophilus spp., Prevotella spp., Actinomyces spp., Bifidobacterium spp. and Veilonella spp. were most often cultured. 66.6% of patients had IgG or IgA antibodies, in contrast, to the control group in which 60.0% and 44.0% of examined blood donors had IgG and IgA antibodies respectively. COPD patients were more frequently positive for specific anti-C. pneumoniae antibodies than the healthy donors (p = 0.003). The difference in a seropositivity rate of specific IgA and IgG antibodies was significant (p = 0.00002 and p = 0.003 respectively). Bronchoalveolar lavage of patients suffering from COPD can be contaminated with high number of aerobic and anaerobic bacterial species, and immunological status of the patients indicated persistent infection caused by C. pneumoniae more often than in controls. PMID- 12632662 TI - [Amplification reaction to bacterial DNA for diagnosing infective endocarditis]. AB - The aim of this study was to evaluate the usefulness of broad-range bacterial PCR in infective endocarditis of bacterial etiology, and to determine its specificity and sensitivity. Twenty five blood samples were taken for analysis from patients with infective endocarditis and acquired valvular heart disease. Infective endocarditis was diagnosed according to Duke criteria. There were two control groups consisting of patients with acquired valvular heart disease: 10 patients with urinary tract infection and 15 patients without. Three different primer pairs for the region of the gene coding for 16S rRNA were tested, to find the most specific one. The highest specificity was found for F/R primers, as the relevant amplified PCR product was present in every blood sample with infective endocarditis, and also in 4 out of 10 patients with urinary tract infection. Broad-range PCR in bacterial endocarditis is a fast, sensitive and inexpensive tool for the detection of bacteria, but it is far more prone to contamination than species specific-PCR. However, in controlled conditions it may be valuable in the identification of non-specific infection allowing for a more rapid clinical diagnosis of endocarditis. PMID- 12632663 TI - [Presence of fungi in stool of children]. AB - A total of 258 children were tested for the presence of fungi in stool. One group consisted of 148 children with non-specific gastrointestinal tract disorders while the other was a group of 110 asthmatics. A quantitative method of enzymatic and mechanical homogenisation was used. The findings were divided into three ranges as follows: < 10(3), 10(3)-10(5), > 10(5) fungal cells in one gram of stool. The number of > 10(5) fungal cells in one gram of stool was considered as pathogenic and requiring treatment. Such a number of fungi in stool was found in 48.1% of children in the first group and in 35.9% in the second one. However, the percentage of fungal presence was higher in the group of asthmatics (83.6% vs. 70.3%). Candida albicans considerably outnumbered the remaining fungal species in the isolates. It was found out that other than C. albicans Candida species were more resistant to the antifungals. PMID- 12632664 TI - An info-tech disconnect. Even as groups such as Leapfrog push IT as an answer to quality issues, doctors and executives say, 'not so fast'. AB - The future is now. A new wave of information technology is gaining wide support outside the hospital industry as a solution to medication errors and patient safety lapses. But in our survey of information system trends, most firms say they aren't operating or launching a physician order-entry system for drugs or treatments. Consultant Steven Roth, left, says recent examples of disasters have warded off many healthcare executives. PMID- 12632665 TI - Medicare to require reports detailing patient satisfaction. AHRQ creating standard patient-experience survey; hospitals will be collecting info as early as September. PMID- 12632666 TI - Rx for physicians. Bush's plan may safeguard docs from more cuts. PMID- 12632667 TI - Over the line? Feds watch doc walkouts with antitrust concerns. PMID- 12632668 TI - Insurance assistance. Feds say hospitals can subsidize docs in some cases. PMID- 12632669 TI - Bouncing back. Stanford Hospital rings up $12.5 million profit. PMID- 12632670 TI - Raising the bar. HCA accepts strict governance code to settle shareholder lawsuit. PMID- 12632671 TI - It gets political. Technology's role in many health policy issues means CIOs must enter the fray. PMID- 12632672 TI - And now, live from your lower intestine...capsule endoscope proves to be a breakthrough tool for gastrointestinal diagnoses, with the pill often removing the need for some painful procedures. PMID- 12632673 TI - Can cold cuts kill? PMID- 12632674 TI - A major league loss. PMID- 12632675 TI - A miracle denied. PMID- 12632676 TI - Health service use in the Winnipeg Regional Health Authority: variations across areas in relation to health and socioeconomic status. AB - The use of healthcare services in Winnipeg is examined to determine whether groups who appear to have a higher need for medical care actually get more care. Despite universal coverage, considerable variation in service use rates exists. Most of the basic healthcare services are provided in accordance with need as measured by premature mortality rates. Nevertheless, visits to specialist physicians, a variety of high profile procedures, and screening and preventative services appear not to be provided in accordance with need. PMID- 12632677 TI - Long stay patients in Winnipeg acute care hospitals. AB - Risk factors for long-stays (more than 30 days) in Winnipeg hospitals were identified, including sociodemographic, illness, and system characteristics. The largest determinant of length of stay was discharge destination; discharge to a nursing home increased length of stay by 173% and 89% for medical and surgical patients, respectively. Hospital of stay also made a big difference for patients discharged to nursing homes. Only 13% of long-stay patients were discharged to a nursing home; 52% were discharged home. Other factors which were associated with an extended length of stay included, hospitalization for stroke, cognitive impairment, rehabilitation therapy, PEG tube insertion, and inhospital fall. PMID- 12632678 TI - Tonsillectomy rates for Manitoba children: temporal and spatial variations. AB - This study compared tonsillectomy rates for Manitoba children across geographic regions and over time. Shortly after the publication of clinical guidelines for tonsillectomy, provincial rates of this procedure dropped by over 25% between 1994/95 and 1996/97. By 1998/99, rates for non-Winnipeg children had increased to pre-guideline levels, whereas the rates for Winnipeg children remained lower. Significant regional variation existed in all years examined, suggesting that quality of care remains an issue for this procedure. PMID- 12632679 TI - Assessing the performance of rural hospitals. AB - This study developed population-based and hospital-based indicators to examine the performance of Manitoba's 68 rural hospitals. Analyses of the indicators revealed considerable differences in the populations served and their use of rural hospital services. Hospital type was also an important factor for performance. The rural hospital indicators would be useful to hospital planners and regional policy makers for comparison purposes and for highlighting issues that need to be addressed. PMID- 12632680 TI - Trends in health and healthcare utilization in Manitoba. AB - Trends in health status and healthcare utilization were examined for regions of Manitoba from 1985 to 2000. While the provincial premature mortality rate decreased, the difference between the northern and southern regions increased. Hospital admissions remained stable despite major bed closures and an aging population; a decrease in hospital days per capita was observed in all regions. Physician contact rates also remained constant despite a 40% increase in the number of seniors. PMID- 12632682 TI - The relative cost of inpatient care in Manitoba's largest hospitals. AB - Comparative information from 1991/92 to 1997/98 regarding an indicator of the relative financial performance of teaching, urban community and major rural hospitals in Manitoba is presented. Results suggest that the acuity and complexity of patients in teaching and urban community hospitals was similar in 1997/98, and that teaching facilities had higher inpatient care costs in all time periods, even after accounting for differences in case-mix, physician remuneration, capital expenditures and the direct costs of teaching programs. PMID- 12632681 TI - Comparing apples to apples: the relative financial performance of Manitoba's acute care hospitals. AB - This paper presents comparative financial ratios that can be adopted by health system administrators and policy analysts to begin to evaluate the performance of acute care hospitals. We combined financial, statistical and clinical information for 73 acute care hospitals in Manitoba for fiscal 1997/98 to calculate 15 indicators of financial performance. Our findings suggest that there is variability between hospital types in their average costs per weighted case, cost structure and financial performance. PMID- 12632683 TI - Patients hospitalized for medical conditions in Winnipeg, Canada: appropriateness and level of care. AB - A medical record review of patients hospitalized for medical conditions in Winnipeg, Canada during 1998/99 was completed using InterQual utilization review instruments. On admission, 95% of patients required the services provided in an acute care facility. Fifty-eight percent of days in hospital following the day of admission required an acute care setting, and 42% required an alternate level of care. Our results suggest there is room for treating more acute patients within the existing system. PMID- 12632684 TI - Seasonal patterns of hospital use in Winnipeg: implications for managing winter bed crises. AB - This study examined whether Winnipeg hospitals experience predictable "high volume periods" in order to determine whether hospital overcrowding might be anticipated and, therefore, avoided. We found that high-volume periods among medical patients occurred during all but one year between 1987 and 1998. Most high-volume periods occurred during influenza seasons. Preventing such recurrent bed pressures requires a multi-faceted approach, involving preventive efforts to reduce hospital admissions (influenza vaccination) and alternatives to managing the hospital system. PMID- 12632685 TI - Working within boundaries: a patient-centered approach to lifestyle change. PMID- 12632686 TI - New eating and physical activity targets to reduce chronic disease risk factors. PMID- 12632688 TI - Special considerations for the education and management of older adults with diabetes. PMID- 12632687 TI - OASIS and beyond: improving outcomes for home health patients. PMID- 12632689 TI - Diabetes self-management in African Americans: an exploration of the role of fatalism. AB - PURPOSE: This study was conducted to explore the concept of fatalism in relation to diabetes self-management behavior in African Americans with type 2 diabetes. METHODS: Participants (n = 39) were recruited from a clinic sample of African Americans with type 2 diabetes. Seven focus groups were conducted; the sessions were recorded, transcribed, and analyzed to identify themes related to fatalism and diabetes self-management. The ISAS paradigm (individual, symbols, audience, situation), a social psychology theory, provided the theoretical framework for the study. RESULTS: Four dimensions of fatalism were identified: the meaning of diabetes, the illness experience, the individual's coping response, and the individual's religious and spiritual beliefs. For the participants in this study, fatalism seemed to characterize the nature of the interaction between the individual with diabetes and others, the meanings they attached to such interactions, and the decision to adopt an effective or ineffective diabetes self management behavior. CONCLUSIONS: Fatalism was associated with diabetes self management in African Americans with diabetes and was multidimensional in this population; the construct appeared to differ conceptually from the perspective of current measures. PMID- 12632690 TI - Insights from older adults with type 2 diabetes: making dietary and exercise changes. AB - PURPOSE: The purpose of this research was to identify factors that affect the nutrition and exercise behaviors of persons over the age of 55 with type 2 diabetes. METHODS: Focus groups were conducted using a validated focus group interview guide to determine primary health concerns and health behaviors, favored learning modalities, barriers to learning, food preferences, and exercise preferences. RESULTS: The following major themes and subthemes were identified: some risk factors for diabetes and heart disease seem more salient than others; perceived susceptibility for serious outcomes of diabetes can occur through vicarious learning; willpower, often obtained through a belief in God, is necessary for successful behavior change; effective modification of behavior and building self-efficacy starts with small steps; and intrinsic reinforcement is necessary for behavior change. CONCLUSIONS: These data were used to identify strategies and messages to enhance adherence to nutrition and activity recommendations for persons with type 2 diabetes and accompanying cardiovascular risk factors. PMID- 12632691 TI - Stage of change advancement for diabetes self-management behaviors and glucose control. AB - PURPOSE: This study was conducted to evaluate whether patients with type 2 diabetes who participated in diabetes education advanced through stages of change for self-management behaviors and to determine if movement was related to glucose control. METHODS: A cohort of 428 patients with type 2 diabetes participated in a traditional diabetes education program in a large urban center in the Southwest. The sample was predominantly female with less than a high school education, a mean age of 52 years, and a mean duration of diabetes of 7 years. Two interviews were conducted approximately 9 months apart, at 1 to 4 weeks before the educational program and at 6 months after completing it. Blood specimens were collected at each interview to measure hemoglobin A1C (A1C) levels. RESULTS: Most of the patients advanced 1 or more stages of change for at least 1 self management behavior. Those with diabetes for less than 2 years were significantly more likely to advance at least 1 stage of change for diet and exercise than those with diabetes for more than 2 years. Such advancement was significantly associated with a decline in A1C. CONCLUSIONS: Patients with type 2 diabetes who participated in diabetes education advanced through stages of change for self care behaviors. The intervention was more effective for those with a shorter duration of diabetes. PMID- 12632692 TI - Condom use, pregnancy, and STDs in adolescent females with and without type 1 diabetes. AB - PURPOSE: The purpose of this secondary analysis was to describe and compare adolescent females with and without diabetes in terms of use of condoms, pregnancy outcomes, and sexually transmitted disease (STD) outcomes. METHODS: Data from a multisite, case-controlled, telephone survey study were used in a secondary analysis to evaluate the parameters stated in the purpose. RESULTS: The 87 females with type 1 diabetes and 45 nondiabetic females who participated were 16 to 22 years old. Most were Caucasian, in high school, and lived with their parents. No demographic differences were noted between the groups. Of those sexually active, for both groups, the most common contraceptive method was male condoms. Four females with diabetes and 2 without diabetes reported an unplanned pregnancy. Three females with diabetes reported an STD infection (chlamydia or trichomonas). Five females with diabetes and 4 nondiabetic females had abnormal pap tests. CONCLUSIONS: Adolescent females with and without diabetes engaged in less than optimally protected sexual activity, which increased their risk for unplanned pregnancies and STD infections. Further research is needed of the incidence, treatment, and short-term and long-term complications of STDs in adolescent females with diabetes. PMID- 12632693 TI - A survey of the perceptions, knowledge, and use of A1C values by home care patients and nurses. AB - PURPOSE: The purpose of this study was to provide baseline information on the perceptions, use, and knowledge of hemoglobin A1C (A1C) values among home healthcare nurses and patients. METHODS: A convenience sample (44 patients, 26 nurses) from a Medicare-certified home care agency was surveyed using an investigator-developed demographic and knowledge questionnaire. Data collected from telephone interviews (patients), written questionnaires (nurses), and medical record reviews were analyzed using descriptive methods. RESULTS: Most of the home care nurses correctly identified the American Diabetes Association's recommendation for target A1C values; however, nurses rarely contact the physician to obtain A1C results. Nurses do not routinely teach patients about A1C values and inconsistently use A1C values to plan care for their patients with diabetes. Most patients said that they had never had an A1C test performed, and most did not recall the nurse providing any teaching about A1C. CONCLUSIONS: Education related to target A1C values is needed. Methods to increase the availability of A1C results in the home care setting should be explored. The importance and clinical utility of A1C values need to be more effectively conveyed to both nurses and patients in the home care setting. PMID- 12632694 TI - Search for microsatellite markers associated with water-stress tolerance in wheat through bulked segregant analysis. AB - We used bulked segregant analysis (BSA) to identify microsatellite markers associated with water-stress tolerance in wheat. Two DNA pools (tolerant and sensitive) were established from the selected F2 individuals of crosses between water-stress-tolerant and -sensitive wheat parental genotypes on the basis of the paraquat (PQ) tolerance, leaf size, and relative water content. All three traits were previously shown to be associated with water-stress tolerance on segregating F2 progeny of the wheat crosses used in this study. Microsatellite analysis was then performed on the established DNA pools, using 35 primer pairs that included all of the chromosome group 5 (5A, 5B, 5D) markers, to detect microsatellite fragments that were present, absent, or both in the DNA pools and their parental lines. We identified one microsatellite fragment that was present in tolerant parent wheat and the tolerant bulk but absent in the sensitive parent wheat and sensitive bulk. We then followed the segregation of this marker in the tolerant F2 individuals. Use of this marker may significantly enhance the success of selection for PQ- and water-stress-tolerant genotypes in wheat breeding programs. PMID- 12632695 TI - Culture-induced variation in plants of Coffea arabica cv. caturra rojo, regenerated by direct and indirect somatic embryogenesis. AB - Amplified fragment-length polymorphism (AFLP) was used to evaluate the stability of DNA in regenerated plantlets of Coffea arabica obtained by direct (DSE) and indirect somatic embryogenesis (ISE). Cluster analysis using the unweighted pair group method (UPGMA), showed no specific grouping pattern related to the type of embryogenesis. These results suggest that the somatic embryogenesis (SE) process has a mechanism for the selection of normal and competent cells. Bulked DNA from regenerated plants obtained by DSE and ISE, and from the mother plants, was used to characterize specific AFLP fragments associated with each SE process. Twenty three primer combinations were tested. A total of 1446 bands were analyzed, with 11.4% being polymorphic and 84% being specific for regenerated plants. Furthermore, specific bands were detected for DSE, ISE, and the mother plants. These results indicate that the SE process induces rearrangements at the DNA level and demonstrates discrepancies between the mechanisms involved in each SE process. Coffea arabica breeding programs that involve DSE and ISE can use AFLP as an additional tool for assessing DNA stability. PMID- 12632696 TI - Detection of universal variable fragments as markers for genetic studies. A novel technology for DNA fingerprinting. AB - A novel DNA technology enables the detection of universal variable fragments (UVF), thus revealing genetic variation without a priori sequence information. The detection of UVF markers is based on two amplifications of genomic DNA with the polymerase chain reaction. In the first amplification, two short oligonucleotide primers produce a large number of fragments. One primer is based on a microsatellite sequence, whereas the second primer can have any sequence. In the second amplification, the length of the primers is increased in order to decrease the number of amplicons. This enables the selection of polymorphic fragments. Restriction digestion can be used to further increase the number of polymorphisms. Until now, we have demonstrated UVF in several different species. In addition, with the present study we have contributed to the linkage map of the rabbit by localizing 11 UVF markers on different linkage groups. Mendelian inheritance was shown in this linkage study through a backcross of two inbred rabbit strains. The power of the UVF technique is based on the selection for microsatellite variation in combination with the detection of single-nucleotide polymorphisms. UVF thus offers the possibility of increasing the clustering of markers and localizing genes in species for which sequence information is either not present or only scarcely present. PMID- 12632697 TI - Ethical issues raised by genetic testing with oligonucleotide microarrays. AB - Because genes and alterations within them determine the identity, characteristics, and inheritance of every individual, the application of genetic science to humans has long been surrounded by apprehension, controversy, and real or perceived potential for abuse. Crude eugenics practices of the past now find a theoretical rebirth and transformation through the use of modern molecular genetic technologies for mutation detection, predictive and prenatal diagnosis, and, ultimately, gene replacement. The advent of oligonucleotide microarray analysis, in which hundreds or thousands of genes and mutations can be tested in parallel, offers tremendous promise for more accurate, sensitive, and efficient genetic testing. At the same time, however, this powerful technology dramatically increases the number and scope of ethical concerns accompanying each individual test request. This article considers the evolution and implications of these concerns, from the initial ordering of a microarray test by the physician to such issues as informed consent, privacy, confidentiality, clinical utility, discrimination, stigmatization, ethnic and population impact, and reimbursement. PMID- 12632698 TI - Bioinformatics methods to predict protein structure and function. A practical approach. AB - Protein structure prediction by using bioinformatics can involve sequence similarity searches, multiple sequence alignments, identification and characterization of domains, secondary structure prediction, solvent accessibility prediction, automatic protein fold recognition, constructing three dimensional models to atomic detail, and model validation. Not all protein structure prediction projects involve the use of all these techniques. A central part of a typical protein structure prediction is the identification of a suitable structural target from which to extrapolate three-dimensional information for a query sequence. The way in which this is done defines three types of projects. The first involves the use of standard and well-understood techniques. If a structural template remains elusive, a second approach using nontrivial methods is required. If a target fold cannot be reliably identified because inconsistent results have been obtained from nontrivial data analyses, the project falls into the third type of project and will be virtually impossible to complete with any degree of reliability. In this article, a set of protocols to predict protein structure from sequence is presented and distinctions among the three types of project are given. These methods, if used appropriately, can provide valuable indicators of protein structure and function. PMID- 12632699 TI - Methods for the molecular analysis of cancer. An overview. AB - Cancer arises as a result of complex and interacting abnormalities. However, over the past 20-25 yr numerous technological advances in molecular biology have led to a dramatic increase in the identification of the molecular processes involved in the development of cancers. The analysis of these molecular changes can be done on different levels: at the DNA or RNA level, or by assessment of posttranscriptional events. This overview discusses the merits of different methods for the analysis of DNA such as SSCP or DGCE. The exciting methods of RNA expression analysis using oligo or cDNA gene chips are also discussed. The importance of methods to analyze posttranscriptional or the effect that altered telomerase activity or methylation status contributes to the phenotype of the cancer cell is emphasized. These techniques will contribute to a better understanding of cancer initiation and progression and will eventually lead toward the development of new molecular-targeted drug therapies. PMID- 12632701 TI - Small group learning in medical education: a second look at the Springer, Stanne, and Donovan meta-analysis. AB - All in all, the evidence is not convincing. Only four of the nine randomized studies used the conventional small-group learning paradigm and qualify as studies of small-group learning, which are relevant to medical education. The results of one of the four are impossible to interpret because of the involvement of the investigator in teaching and test construction. The three remaining studies showed no effect, a negative effect, and a positive effect, respectively. The nonrandomized studies failed to establish the comparability of the groups. The evidence does not support the authors' call for "more widespread implementation of small-group learning in undergraduate SMET". Small-group learning has not been shown to support the acquisition of content any better [or worse] than large-group learning. In medical education, small-groups are employed in large part to develop team work skills, communication skills, and peer- and self-assessment skills. But these outcomes are not addressed in this meta analysis. More seriously, our rereading of these studies raises general concerns about meta-analysis in education, which have important implications for evidence based medical education. The meta-analysis under discussion at first appeared to be just the kind needed to guide an evidence-based educational enterprise. However, a closer look revealed both what is lacking in the meta-analysis and some of the ways educational research and reporting need to be changed if anything like evidence-based education is ever to become a reality. At the least, study design must be clearly described. In addition, if the design is nonrandomized, the groups should be described in sufficient detail to allow a meaningful interpretation of the role of preexisting differences on the outcome measures. (This is why we limited our discussion here to the randomized studies.) Also, effect-size measures should be reported for all comparisons that bear on the impact of the intervention, including preexisting differences. Reporting significance is not enough. This shows only whether sampling error can be ruled out (with a low probability of error, p < .05) as a possible explanation of the connection between the intervention and the outcome. The effect can still be trivial and the comparisons confounded. In addition, descriptions of the actual educational interventions employed need to be more comprehensive and precise. For the most part, the papers would have been strengthened by providing more information for replicating the studies and for deciding which should be included in a given meta-analysis. Perhaps most seriously, our rereading of these studies makes us wonder about the possibility of meaningfully synthesizing the results of educational studies, given their idiosyncrasies and their many extraneous, uncontrolled factors. The conclusions from most educational studies, then- whether randomized or not--must be highly qualified, with explicit warnings about preexisting differences and other confounding factors that plausibly account for the study results. However, these narrative qualifications do nothing to adjust the effect-size measures, which are typically pooled or synthesized across studies--confounds and all. The idiosyncrasies of the studies seem to preclude a blanket qualification that can be applied conceptually across the collection of studies to arrive at a sound conclusion from the synthesis. In brief, the meta analysis considered here does not support the application of small-group learning in medical education and it raises questions about meta-analysis in education with implications for evidence-based education. PMID- 12632700 TI - Improved method for generating microarray probes using submicrogram amounts of total RNA. AB - The ability to identify disease-associated genes using microarray technology is dependent on isolating high-quality total RNA from the diseased tissue under study. However, it is not always possible to obtain large amounts of affected tissue from patients and therefore the quality and yield of RNA may be compromised. This protocol described a procedure for generating superior microarray probes by amplifying RNA sequences via successive rounds of in vitro transcription (IVT) reactions. This improved procedure utilizes a 9-mer primer to generate the IVT template, which is able to recapitulate the size ditribution of the original isolated RNA sample. PMID- 12632702 TI - Development and implementation of an objective structured teaching exercise (OSTE) to evaluate improvement in feedback skills following a faculty development workshop. AB - BACKGROUND: Faculty development programs focusing on teaching have become widespread. PURPOSE: Despite the popularity of such programs, evidence as to their effectiveness is limited. This article reports on the development of an objective structured teaching exercise (OSTE) and its pilot implementation in an evaluation of a faculty development program module. A written test intended to measure feedback skills was also developed and pilot tested. METHODS: A separate sample, pretest-posttest design was used to pilot test both instruments. RESULTS: The results showed some evidence of significant differences between groups tested preworkshop and postworkshop. Higher scores were observed for the posttest group compared to the pretest group only for OSTE items focusing on prioritizing and limiting the amount of feedback given at one time and on action planning. CONCLUSIONS: Results suggest that an OSTE may be sensitive to changes in preceptor skill level for skills that are relatively easy to incorporate immediately into practice. Lack of differences in other skill areas may be due to lack of sensitivity of the measure or to need for practice and reflection before changes in performance on other feedback skills are evident. PMID- 12632703 TI - Student understanding of the relationship between the health professions and the pharmaceutical industry. AB - BACKGROUND: Pharmaceutical sales representatives and direct-to-consumer advertising may influence physician practices, particularly prescribing. Identifying the relevant knowledge and attitudes students possess about the pharmaceutical industry may help professional curricula address these influences. PURPOSES: To assess knowledge and attitudes toward pharmaceutical industry marketing, ethical principles guiding drug company interactions, pharmaceutical sales representatives as a source of drug information, and confidence level in addressing consumers seeking a prescription from a direct-to-consumer advertisement among senior-level medical, PharmD, and nurse practitioner students. METHODS: A cross-sectional survey design was used to assess student knowledge and attitudes of four domains associated with the pharmaceutical industry. RESULTS: Significant deficiencies were noted in student knowledge of pharmaceutical marketing expenditures, professional ethics regarding interactions with drug companies, and accuracy of drug information from sales representatives. CONCLUSIONS: Health professional students' knowledge and attitudes toward the pharmaceutical industry are formed prior to graduation. Professional curricula must address the influences of sales representatives before postgraduate training. PMID- 12632704 TI - Does being a clerkship director benefit academic career advancement: results of a national survey. AB - BACKGROUND: Changes in academic medicine have left clerkship directors (CDs) anxious about their career pathway, because clerkship administrative efforts may detract from other activities. PURPOSE: The Clerkship Directors in Internal Medicine (CDIM) asked members about benefits of being a CD or CDIM membership toward career development. METHODS: Responses were on 1-5 Likert scales with 5 (strongly agree). Background and demographic issues were analyzed for associations with the career benefits statements. RESULTS: The response rate was 75% (n = 92). Mean agreement with CD benefit was 4.2 (SD = 0.82) and CDIM membership 3.8 (SD = 0.95). Eighty-one percent and 58% of CDs agreed with the respective statements. Significant predictors of CD benefit were CD and coordinator salary support, years as CD, and receiving a university teaching award. Structured discussions of expectations strongly predicted perceiving CDIM benefit. CONCLUSIONS: Most CDs agreed that their CD role and CDIM benefited their careers. Salary support and clearly defining expectations may increase the likelihood of perceiving benefit. PMID- 12632705 TI - Effectiveness of report cards based on chart audits of residents' adherence to practice guidelines on practice performance: a randomized controlled trial. AB - BACKGROUND: Medical record audits have been used to provide physicians with feedback about their compliance with preventive health and disease management recommendations. PURPOSE: To determine if report cards summarizing medicine residents' preventive health and disease management practices can be used as a feedback tool to improve practice performance. METHODS: Randomized, blinded, controlled study of 44 internal medicine residents using an individualized 78 item report card based on outpatient record audits. RESULTS: Four hundred ninety seven charts were retrospectively audited at baseline and 284 charts in follow up. There were no significant differences in baseline performance between the residents in the intervention and control group. There were no differences in performance scores between residents receiving report cards and those who had not in immunizations, counseling, total preventive health, diabetes, hypertension, and total disease management. CONCLUSIONS: Intensive data-based feedback using report cards may not be a successful way to improve ambulatory performance of medical house officers. PMID- 12632706 TI - "My future as a physician"--professional representations and their background among first-day medical students. AB - BACKGROUND: Professional representations orient students toward their future profession and influence their view of what things are important to learn. PURPOSE: To analyze medical students' professional representations and their background. METHODS: One hundred nine 1st-day medical students wrote an essay titled "My work and future as a physician in the 21st century" and filled in a questionnaire about their background. The essays were content analyzed to identify the dimensions of their professional representations. RESULTS: Four dimensions of representations emerged: impersonal-personal, foreclosed reflective, traditional-transformative, and idealistic-realistic. The professional representations were associated with the students' gender, previous experiences of working and studying, their sources of information about the medical profession, and the time of career choice. CONCLUSIONS: Our study revealed different subgroups of medical students in view of their professional orientation. The relevance of this finding for the recruitment and educational practices of medical schools is discussed. PMID- 12632707 TI - Morning Report: an analysis of curricular content and comparison to national guidelines. AB - BACKGROUND: Morning Report is a long-standing tradition for internal medicine residency programs, but format and content vary greatly and have not been compared to any accepted curriculum standards. The Federated Council for Internal Medicine (FCIM) Task Force on the Internal Medicine Residency Curriculum has published curriculum guidelines for residency programs. PURPOSE: We compared our institution's Morning Report topics with FCIM curriculum guidelines to determine the feasibility and validity of these guidelines in a residency program. METHODS: A review was performed of all Morning Report conferences over 3 years. RESULTS: With the exception of general internal medicine topics, between 60% to 86% of all FCIM organ and system competency focus areas were the subject of one or more of 583 consecutive Morning Reports. CONCLUSIONS: The content of our Morning Report conferences closely paralleled FCIM guidelines, suggesting that the FCIM guidelines may be feasible and valid. These guidelines can provide guidance for Morning Report content. PMID- 12632708 TI - The Oregon Center for Complementary and Alternative Medicine career development program: innovation in research training for complementary and alternative medicine. AB - BACKGROUND: The burgeoning use of complementary and alternative medicine (CAM) highlights the need for high-quality research to discriminate between those approaches based on positive anecdotes or polished marketing and those that are consistently effective and safe. DESCRIPTION: To help meet this need, the Oregon Center for Complementary and Alternative Medicine has developed an innovative program to train researchers. The training program includes an individual mentoring and training plan, group mentoring, a clinical research class, proposal development, completion of an actual study, and journal club participation. EVALUATION: After 2 years the fellows have submitted 18 grants, presented 4 posters, submitted 7 articles, developed 2 treatment protocols, and earned 1 master of public health degree and 1 graduate certificate. CONCLUSIONS: This program helps experienced CAM practitioners quickly adopt a research perspective. We encourage other career development programs to extend training in research methods to individuals possessing a profound understanding of CAM. PMID- 12632710 TI - Medical student perceptions of factors affecting productivity of problem-based learning tutorial groups: does culture influence the outcome? AB - BACKGROUND: The popularization of problem-based learning (PBL) has drawn attention to the motivational and cognitive skills necessary for medical students in group learning. This study identifies the effect of motivational and cognitive factors on group productivity of PBL tutorial groups. DESCRIPTION: A self administered questionnaire was completed by 115 students at the end of PBL tutorials for 4 themes. The questionnaire explored student perceptions about effect of motivation, cohesion, sponging, withdrawal, interaction, and elaboration on group productivity. We further analyzed (a) differences in perceptions between male and female students, (b) effect of "problems," and (c) effect of student progress over time on group productivity. EVALUATION: There were linear relations between a tutorial group's success and the factors studied. Significant differences were noted between male and female student groups. CONCLUSIONS: Students and tutors need to recognize symptoms of ineffective PBL groups. Our study emphasizes the need to take into account cultural issues in setting ground rules for PBL tutorials. PMID- 12632709 TI - Medical Students for Choice: origins, current orientations, and potential impact. AB - BACKGROUND: Medical Students for Choice is a U.S.-based organization concerned with the shortage of abortion providers, the lack of abortion education in medical schools, and violence against abortion providers. Created in 1993, MSFC has 110 chapters in North America with 7,000 student affiliates. DESCRIPTION: The rapid spread of MSFC chapters since its creation prompted the Northeastern Ohio Universities College of Medicine's chapter to undertake group interviews of 40 medical students affiliated with 7 MSFC chapters in 4 midwestern states in the United States. EVALUATION: Interview questions were developed to identify students' motivations for affiliation with MSFC, how abortion was currently addressed in the medical curriculum and where it should be, how students think abortion providers are perceived by the medical community, and how they plan to integrate their commitment to reproductive choice in their future practice. CONCLUSIONS: Not all students affiliated with MSFC will become abortion providers, but they are politically enthusiastic and committed to the inclusion of abortion in medical training and to legitimizing and normalizing its provision. More research tracking this group across time and geography will add much to understanding abortion services in the United States and what part the medical curriculum plays in educating students to provide comprehensive reproductive health services for women. PMID- 12632711 TI - Nurse recruitment: a waste of postage, paper, and people. PMID- 12632712 TI - Recruitment and retention strategies: a magnet hospital prevention model. PMID- 12632713 TI - Nursing intellectual capital: a theoretical approach for analyzing nursing productivity. PMID- 12632714 TI - Market power: an interview with Miles Snowden of Delta Air Lines and the Leapfrog Group. Interview by Alison P. Smith. PMID- 12632715 TI - Managed care education for nurses: practices and proposals. PMID- 12632717 TI - Using IT to improve clinical teamwork and communication. PMID- 12632716 TI - Cholesterol management in the ambulatory setting for the underserved. PMID- 12632718 TI - A recruitment and retention program that works! PMID- 12632719 TI - Is Six Sigma the answer for nursing to reduce medical errors and enhance patient safety? PMID- 12632720 TI - Compliance leadership: the 17th century model that doesn't work. AB - The world of health care has changed. We can't operate on 17th century models and be successful. We don't have to argue for the movement to committed, inspired leadership models, and the death of compliance leadership. There is abundant research to document this is the way we must go. In reality, we have a moral obligation to provide the kind of work environment that provides the meaningful work that Maslow (1998) tells us makes life meaningful. No one has the right to make people miserable at work because we have failed to create the cultures that create commitment, inspiration, and transformation for our patients, their families, and our staff. It is unfortunate that in times of staff shortages, this message is heard louder. We should be equally committed no matter what the situation, because it is the right thing to do. We do not have the right to be abusive to others in any context. PMID- 12632721 TI - Supporting the nurse manager to improve staff nurse retention. PMID- 12632722 TI - Health care policy or politics--which prevails in 2003? PMID- 12632723 TI - AIDS. A crisis get its due. PMID- 12632724 TI - An old enemy is back. PMID- 12632725 TI - [On the mechanism of Ca2+<--> Mg2+ interactions in dental enamel]. PMID- 12632726 TI - [The ultrastructure of gingival epithelial prickle layer cells in patients with chronic catarrhal gingivitis concomitant with connective tissue dysplasia]. AB - Patients with chronic gingivitis and connective tissue dysplasia developed more marked dystrophic process in the gingival epithelium than those with chronic gingivitis without general somatic diseases. These dystrophic processes substantially increase the barrier properties of the gingival epithelium. PMID- 12632727 TI - [Levels of amino group nitrogen and imidazole compounds in human oral fluid]. AB - The procedures of definition amino nitrogen (sum amino acids) and imidazole compounds in human oral fluid are described. Amino nitrogen were determined by the ninhydrin method modified by the authors, imidazole compounds by the Pauli test. A total of over 20 patients with the relatively good dental status were surveyed. The level of amino nitrogen showed noticeable fluctuations (2.01 +/- 1.24 mmol per ml of oral fluid), whereas the content of imidazole derivatives was more stable (0.71 +/- 0.19 mmol/ml). Oral fluid samples displayed an in vitro high proteolytic activity, which explains contradictory results in the determination of the levels of amino nitrogen. PMID- 12632729 TI - [Dental caries resistance under conditions of a space flight]. AB - The maxillodental status of aeronauts attracts special interest of scientists and causes apprehensions of physicians and scientists of countries with aeronautic industry because of long duration of space missions. Some scientists consider that the realization of mission to Mars can lead to development of multiple dental caries in aeronauts. The aim of this study was to study the effects of space mission factors on the resistance of hard dental tissues and remineralizing activity of the saliva in a simulation experiment. The results demonstrated dynamic changes in the studied parameters under the effect of space mission factors; based on these results, we determined the criteria for selection of candidates for prolonged experiments and space missions. PMID- 12632728 TI - [Study of the characteristics of biological membranes used in osteoplasty with the aim of purposeful tissue regeneration on a model of continuous bone marrow cultures]. AB - The biological materials Parodoncol and Hapcol (Polystom Firm) used in osteoplasty were studied in continuous mouse bone marrow cultures. Culturing in continuous bone marrow cultures showed growth and development of stromal and hemopoietic cells on Hapcol and Parodoncol. The materials caused no negative reactions and failed to inhibit bone marrow cell growth. The number of cells gradually increased, hemopoiesis was normal, and young and mature granulocytes formed. This method of culturing proved to be highly effective for evaluation of the characteristics of biological membranes and other composite materials. PMID- 12632730 TI - [The reaction of pulp vessels to various adhesive systems]. AB - The reaction of pulp vessels to various adhesive systems was studied: qualitative and quantitative characteristics of pulse curves reflecting the pulp blood filling were studied. Study of the reaction of pulp vessels showed that acetone containing adhesive systems immediately caused a constrictive reaction, while use of an acetone-free adhesive system led to the minimum changes in the functions of pulp vessels. PMID- 12632731 TI - [Eryxine effect on hydroxyproline metabolism in patients with chronic periodontitis]. AB - The paper gives the results of the use in clinic of the new preparation eryxine and its effects on the concentration of hydroxyproline in the saliva and serum in patients with chronic periodontitis. PMID- 12632732 TI - [Osteoplasty with membranes: indications for use, probable errors, and violation of the action principle of purposeful tissue regeneration]. PMID- 12632733 TI - [Maintenance of anesthesia and intraoperative safety in dental implantology]. AB - Most patients who need dental implantation suffer from somatic diseases. Therefore, an anesthesiologist's participation is highly desirable, since he supports the stability of vital functions. The authors used mixed anesthesia involving traditional field-block and general anesthesia. By choosing doses of anesthetics, the authors achieved ataraxia which preserved the minimum contact with a patient and a complete analgesia, which gave rise to psychoemotional stability. PMID- 12632734 TI - [Use of X-ray techniques in dental implantology]. AB - This paper deals with some aspects of using radiographic methods in dental implantology. Analysis of the published data and their own findings have brought the authors to the conclusion that the most widely used method involved is a panoramic technique; however, CT scan is required in some cases demanding accurate measurements; which, at the stage of preoperative planning, enables us to specify surgical indications and choose the optimal form of implants. PMID- 12632735 TI - [Efficacy of treating denture-induced stomatitis by electrically activated aqueous solutions]. AB - The authors investigated the oral mucosa in 35 patients, who were divided into four groups: Group 1 (n = 17) received traditional treatment, Group 1 (n = 18) had the similar treatment supplemented by electrically activated aqueous calcium chloride solutions. The findings have shown that the use of electrically activated aqueous calcium chloride solutions in the complex treatment of orthopaedic stomatitis exert more marked clinical and immunological effects than the traditional treatment (p < 0.001-0.01). PMID- 12632736 TI - [Temporomandibular joint dysfunction in patients with rheumatoid arthritis]. AB - Clinical and instrumental examinations of 120 patients with rheumatoid arthritis (RA) detected a direct correlation between RA stage and severity of dysfunctions and an inverse correlation between the disease stage and angle of the sagittal articular route of the temporomandibular joint. PMID- 12632737 TI - [Special basic material for microwave polymerization of AKR-MV]. AB - Some physical and mechanical properties of the new acrylic material ACR-MW for microwave polymerization, the base material Etacryl-02 ("Stoma", Ukraine) and their analogue Acron MC (GC Lab Technologies Inc., Japan). The findings suggest that the new special denture base material ACR-MW for rapid microwave polymerization (3 min at 100%) possesses the highest bending strength with lower water absorption and solubility. The properties of ACR-MW meet the requirements of ISO 1567. PMID- 12632738 TI - [Registration statistical program for the Rehabilitation Center for Patients with Cleft Lip and Palate in Uzbekistan]. AB - The authors suggest an information computer support within the framework of a complex system for the diagnosis and treatment of patients with cleft lip and palate. This program will ensure operative information exchange for specialists of various spheres. Such information, united in the data base, will become the base for creation of a universal system of complex rehabilitation of patients with cleft lip and palate. PMID- 12632739 TI - [Preoperative bifidum-lactobacterin therapy in children with cleft lip and palate]. AB - Microbiological and immunologic investigations of peripheric blood in 125 children were conducted. They determined a positive role of early correction with eubiotics (bifidum-lactobacterium) in children with congenital cleft lip and congenital cleft palate during 8-9 months before chiloplasty and uraniscoplasty. Using eubiotics results in normalization and restoration of the intestinal microflora, which leads to restoration of cellular and humoral immunity and to decreased rates of accompanying and postoperative complications. PMID- 12632740 TI - [Formation of the network of state institutions of dentistry in different regions of the Russian Federation (preliminary analysis)]. PMID- 12632741 TI - [Dental aspects of infantile cerebral paralysis]. PMID- 12632742 TI - [History of development of the Department of Dental and Maxillofacial Surgery, Russian Medical Academy of Postgraduate Training]. PMID- 12632743 TI - [Role of the vitreous body in vitreoretinal diseases]. AB - Recent research during the last decades revealed evidence for the pathogenic role of the vitreous in many diseases of the retina. After a survey of the surgical anatomy and biochemistry of the vitreous, the author summarises present knowledge of various vitreoretinal disorders. Vitreoretinal disorders are discussed as follows: congenital anomalies, pathologic vitreoretinal adhesions, vitreous opacities, inflammatory diseases, vitreous hemorrhages and cellular infiltration of the vitreous. Special emphasis is placed on vitroretinal disorders of general importance, such as diabetic retinopathy, endogenous (fungal) endophthalmitis, Whipple disease, Crohn disease, sarcoidosis, shaken-baby syndrome, Terson syndrome, metastatic tumours of the choroid, cellular infiltration of the vitreous due to leukaemia or non-Hodgkin lymphoma, and amyloidosis. During the last few years better understanding of vitreoretinal disorders and improved diagnostic and therapeutic methods opened a new era of vitreoretinal surgery. PMID- 12632744 TI - [Toxic shock syndrome caused by pyogenic bacteria]. AB - Case reports and review of the literature. INTRODUCTION: Severe toxic shock syndrome caused by invasive infection with pyogenic bacteria Staphylococcus aureus or group A Streptococcus pyogenes, with high mortality rates in cases of the latter, remained one of the most problematic chapters of critical care medicine to date. AIMS: To give an overview on the epidemiology, clinical manifestations, the complex therapeutical approaches of the syndrome and, on the role and mechanisms of action of bacterial superantigens in the pathophysiological processes as well. METHODS AND RESULTS: Literary data, and some illustrative selected cases demonstrate that, the incidence of TSS shows increasing tendency worldwide and, that otherwise healthy, younger people are the most frequently affected. As for prognosis: early diagnosis and treatment with sufficient radicality are of decisive importance. PMID- 12632745 TI - [Hyperinsulinemia, insulin-like growth factor-I, insulin-like growth factor binding protein-1, and sex-hormone binding-globulin in prepubertal and pubertal girls with premature adrenarche]. AB - Precocious adrenarche is defined as the development of pubic hair before the age of 8 years in girls and 9 years in boys. Pubarche caused premature adrenarche in girls has been considered as a normal variant of pubertal development for years. Recently, it is cleared that premature pubarche can be considered as a marker of increased risk for endocrine and metabolic abnormalities. Precocious adrenarche in affected girls is associated with hyperinsulinaemia and functional ovarian hyperandrogenism during puberty. Authors investigated serum levels of insulin like growth factor-I (IGF-I), insulin-like growth factor-binding protein-1 (IGFBP 1), insulin-like growth factor-binding protein-3 (IGFBP-3), sex hormone binding globulin (SHBG) and levels of insulin during oral glucose tolerance test in 34 girls with premature adrenarche in 38 age- and BMI-matched healthy controls. Affected girls were assigned into prepubertal and pubertal subgroups. It has been shown that hyperinsulinaemia, decrease in IGFBP-1 and increase IGF-I levels may be present in some affected prepubertal patients. In the pubertal group, in addition to hyprinsulinaemia, decreased IGFBP-1 and increased IGF-I levels an attenuated SHBG level was observed. According to the authors, these laboratory parameters may predict endocrine and metabolic abnormalities in later life. The observed correlations support the hypothesis that insulin/IGF system plays role in the pathogenesis of hyperandrogenism in premature adrenarche and in later hormonal and metabolic changes. PMID- 12632746 TI - [Ethical regulation of biomedical research in Hungary]. AB - Hungary joined the Ovideo Treaty (a bioethical health agreement signed by EU countries in 1997), as declared by Law Nr. VI in 2002. In July 1 2002 two departmental orders were enacted: departmental order Nr. [23/2002 (V. 9.)] about the biomedical research in humans and departmental order Nr. [24/2002 (V. 9.)] about the human use and clinical trials of investigational products and the adaptation of Good Clinical Practice (GCP). Both were based on the Health Law. The departmental order on the Medical Research Council [(16/2001 (IV. 28.)] together with these two orders contains the national rules of biomedical research performed in Hungary and also the Hungarian adaptation of various, bioethical principles and directives published by EU or other internationally accepted organisations. According to these regulations from 2002 biomedical research in Hungary could be in agreement with the Oviedo Treaty. Ethical approval and supervision can be obtained from research ethical committees of three types: central, regional and local superimposed upon one another. There are three, central, ethical committees within the frames of the Medical Research Council with national responsibility: Scientific and Research Ethical Committee, Clinical Pharmacological Ethical Committee, Human Reproduction Committee. In some cases regional research ethical committees are entitled to hand out ethical approval and ethical supervision. In those research sites, where the regional research ethical committee is not existent a local committee should be organized for the local ethical control of the research investigations and experiments. This way the ethical requirements and clinical practice by the GCP can be performed in clinical research in Hungary with a special respect to the vulnerable persons possibly involved. The paper gives an overview on recent developments and major ethical principles of the Hungarian biomedical research. PMID- 12632747 TI - [Medical observations in the Gulag (1945-55)]. PMID- 12632748 TI - [Clinical features of parathyroid diseases. 1952]. PMID- 12632749 TI - [Anti-IGE therapy in asthma]. AB - Immunoglobulin E (IgE) is the hallmark of allergic diseases and asthma. Regulating IgE production has been the focus over several years as an important strategy in the treatment of allergic diseases. Recently, nonanaphylactogenic antihuman IgE antibodies have been under clinical evaluation as a therapeutic agent against atopic disease. In asthmatic subjects, the administration of these monoclonal anti-IgE antibody has been shown to reduce plasma IgE levels, reduce early and late phase allergic responses after allergen provocation, improve symptoms and reduce rescue medication. No serious side effects were reported. Thus, the clinical effectiveness of these medications supports the viability of anti-IgE therapy as a potentially effective treatment option for asthma. PMID- 12632750 TI - [Stratification of cardiac risk in unstable angina: value of troponin measurements]. AB - Recent studies have identified patients with unstable angina and increased troponin I or T as a high risk population gaining benefit from adjunctive treatment with glycoprotein IIb/IIIa receptor antagonists and early reperfusion by coronary interventions. PMID- 12632751 TI - [Epidemiologic, clinical and therapeutic aspects of chronic lymphoid leukemia: apropos of 120 cases]. AB - We retrospectively studied 120 cases of chronic lymphocytic leukemia diagnosed between January 1988 and December 1998. The median age of our patients was of 66 years, 75% among them were male. The discovery of the illness was fortuitous in 20% of the cases, the peripheral adenopathy and the splenomegaly were noted respectively in 72 and 48% of the cases. The blood lymphocytosis was on average 51.109/1 with extremes of 5 and 818.109/1. Anemia was noted in 71% of the cases and a thrombopenia in 42%. Fifty patients were classified C stage of BINET and sixty elevated risk according to RAI. The therapeutic attitude was according to patient's age and the CLL stage. Thus, 94 patients received a chemotherapy and a complete or partial response was observed in 58 of the cases. The overall survival at 5 years were 47%. The retained prognostic factors were the stage according to the classifications of BINET and RAI, the thrombopenia and the lymphocytosis blood overhead 100.109/1. PMID- 12632752 TI - [Endometrial resection in the treatment of persistent metrorrhagia]. AB - STUDY OBJECTIVE: To evaluate the efficacy of hysteroscopic endometrial resection such us a surgical alternative to hysterectomy in treating abnormal uterine bleeding unresponsive to treatment with progestogens. METHODS: A retrospective analysis was carried out on 54 patients who underwent endometrial ablation for abnormal uterine bleeding unresponsive to conservative medical management between January 1, 1991, and December 31, 1998. The average patient was 43 years (Range: 36-57 years) and was followed for a mean of 37 months (range 12-80 months). RESULTS: 83.3% of women were satisfied during the study period. The overall amenorrhea rate was 29.62%. Only 16.66% of subjects reported no improvement. Histologic analysis of the endomyometrial specimens revealed hyperplasia in 48.1% of cases, atrophy in 3.7% of cases and adenomyosis in 44.6% of cases. 8 women (14.8%) needed secondary hysterectomy for continued symptoms during a mean follow up of 10 months. Adenomyosis was present in 4 specimens (50%), myomas in 3 specimens (37.5%), and the association in 1 specimen (12.5%). CONCLUSION: Endometrial ablation is a safe and effective hysteroscopic procedure for dysfunctional uterine bleeding, its prognosis depends of several factors, that shows the importance of patients selection. PMID- 12632753 TI - [Prevalence of Helicobacter pylori infection in duodenal ulcer. Data of a prospective study apropos of 78 NSAID-negative patients with duodenal ulcer]. AB - The aim of our study was to evaluate, the prevalence of Helicobacter pylori infection in duodenal ulcer tunisian patients not using non steroidal anti inflammatory drugs. Seventy eight patients having a non complicated duodenal ulcer and not using non steroidal anti inflammatory drugs were enrolled in this prospective study carried out between February 2000 and January 2001. Six gastric biopsies were undertaken in all patients; five for an histologic analysis and one for urease test. A serum sample was taken for a Helicobacter pylori serology. Patients were considered infected if two tests were positive. Helicobacter pylori infection was present in 77 patients (98.7%). Helicobacter pylori infection is still high among patients with a non complicated duodenal ulcer and not using non steroidal anti inflammatory drugs suggesting an empirical Helicobacter pylori eradication therapy in this group of patients. PMID- 12632754 TI - [General medicine and general practitioner. The viewpoint of the teachers at the Department of Medicine of Tunis]. AB - The general medicine remains a same survey subject in countries where the debate began since several years. We have led an investigation by questionnaire close to 50 chosen hospitalo-academic physicians by pull to the fate on the list of teachers of the faculty of medicine of Tunis. The majority of our investigated (90%) consider that the physician general practitioner is the physician of the individual in its totality, the counselor and the family's confidant. The formation that receives the future general practitioner currently doesn't prepare it to assure his role. To form the future physician better general practitioner, we think that it's necessary to define the general practitioner role, to adapt the formation to the role of the general practitioner and the specificity of the general medicine. PMID- 12632755 TI - [A programmed autologous transfusion. A study of 47 patients]. AB - From December 1994 to January 2002, 44 among the 47 patients proposed have been integrated in the protocol of programed autologous transfusion. 18 patients were belonged to the male sex and 26 others belonged to the female one. The average age was 53.2 years (range 15-82 years old). 39 among the patients admitted in the protocol had an orthopaedic pathology. The protocol has associated the teams of surgery, of anesthesia and of transfusion and has occurred in the respect of the regulation (circular 91/2000). The blood taking have led to a significant modification of the hemoglobin rate (average decrease of 2 g/100 ml), of hematocrit (average decrease of 6.1%) and of the rate of platelets (average increase of 29,324 platelets/mm3). On the other hand, a transfusional complement by the concentrated homologous red corpuscles was necessary for 2 patients (2/40). PMID- 12632756 TI - [The dual chamber cardiac pacing. A multicenter study apropos of 353 pacemakers]. AB - 11 cardiac units, among which 5 experienced cardiac pacing are involved in the study. A retrospective analysis of the operated patient is performed. Authors paid particular attention to the following items: indications, etiologies, per operative parameters, complications and programmatic features of the inserted devices. Demographic date: 176 M, 177 F. The mean age of the sample: 58.4 +/- 16.25 years. The post operative complications occurred in 14% of the patients and were favourable evolution, but the presence of an atrial lead generated the half of the reoperations of which incidence was around 4%. (2%) have led to leave the DDD mode. The favorable results, as well at the technical levels as the evaluating level, witness that the tunisian centers have acquired a sufficient knowledge in the field of the conventional pacing and could now born on other problems, out passing the field of the bradycardias. PMID- 12632757 TI - [Contribution of clinical, biological and radiological parameters in the diagnosis of urinary infection localization]. AB - Urinary infection was a problem in pediatrics. Currently the diagnosis is easy but it is hard to diagnosis the localisation of the infection. We have to go fast and precisely to treat correctly this infection and diminich the possibility of developing renal scars. Dimercaptosuccinic acid (DMSA) scintigraphy is a reference exam for detection acute renal lesions. We have realised a prospective study in 29 children presenting urinary tract infection. In whom a DMSA scintigraphy is realised between the one day and 30 days after the infection. We study the sensibility ans the specificity of clinical, biological and radiological parameter for the diagnosis of localisation of the infection. The better parameter of sensibility of upper urinary tract infection is of fever, and the better parameter of specificity is the association of fever, echo Doppler renal and VS. PMID- 12632758 TI - [Risk factors of drug use in school environment in Tunis]. AB - The aim of the study was to estimate the prevalence of addiction and drug use among Tunisian school teenagers as well as the risk factors associated with this conduct. A transversal study of "Knowledges, Behaviors and Practices" (KBP) has therefore been conducted among 353 school teenagers in the area of Tunis by means of an anonymous and self administered questionnary. Pupils' ages range from 12 to 24 with as many boys as girls (51.6% VS 48.4%). Three quarter (3/4) of the pupils have heard of drugs, the cannabis ("zatla") being the most common referred to; nearly 13% of the pupils said they have consumed drug at last once, half among them more then 3 times. The risk factors associated with drug use are the male sex (p = 10-7), tobacco or alcohol consumption (p = 10-8), violent behaviors (p = 10-4) and a confactual situation with parents (p = 10-5). A prevention of the problem should be based on an action over the identified risk factors. PMID- 12632759 TI - [Abdomino-pelvic actinomycosis: a case report]. AB - The actinomycosis is a chronic suppurative granulomatosis disease. It is owed to a bacillus gram positive; actinomyces israelli. The cervical and thoracic localizations are most frequent. The digestive localization represents 20% of cases. It interest very rarely the pelvis and the genital tracts. We bring back the observation of a patient old of 30 years admitted for mass abdominal. To the exam, the patient had a sensibility of the left hypochondriac area and we found a mass of 6 cm of diameter. To the rectal touch, we found a mass in the bag of Douglas. The echography and the computed tomography revealed a collection under the spleen and a pelvic collection. A rectotomy is performed. The bacteriological study isolates actinomyces israelli. The collection under the spleen is drained under radiological control. Actinomyces israelli is also recovered in the pus brought back by the puncture. The patient is treated by Penicillin. The patient had a favourable evolution. No etiology is found at this patient. For this observation, the collection was accessible to a drainage permitting the diagnosis and the treatment of the actinomycosis while avoiding a mutilated surgery. PMID- 12632760 TI - [Coronary reoperation through left thoracotomy without cardiopulmonary bypass: 3 case reports]. AB - With the increasing number of patients who have undergone coronary artery bypass grafting, the incidence of reoperative coronary grafting is also increasing. Reoperative coronary artery bypass grafting is associated with morbidity and mortality rates greater than those of primary coronary operations. Left thoracotomy can provide access for reoperation when repeat median sternotomy is extremely dangerous. Coronary artery grafting on a beating heart and via left thoracotomy can be a good alternative strategy when redo coronary surgery is necessary in the circumflex or left anterior descending territories. PMID- 12632761 TI - [Medical prevention of handicap in Tunisia]. AB - The handicap, phenomenon too often badly discerned, constitute a problem of public health of part his frequency, his gravity and his cost. Interventions aiming to warn the handicap are located to three levels: primary, secondary and tertiary. The measures as the vaccination, the education about health, the hygiene of the setting, the prenatal surveillance, aiming to reduce and even eradicate the transferable illnesses, is applied and integrates in the setting of the different national programs since years 80. The most important are the national program of vaccination and the national program of perinatality. The precocious tracking of deficiencies and their hold in charge is related to centers of cares of basis health, to the regional units of rehabilitation and the school and academic medicine. The sanitary measures of tertiary prevention of the handicap summarize themselves in two types of measures: the rehabilitation and the equipment. The handicap, multi-dimensional phenomenon, require a concerted multidisciplinary action between professionals of health, social business, jurists and community. PMID- 12632762 TI - [Pacemaker infections]. AB - Infectious complications following pacemaker implantation are not common but can be particularly severe. The reported incidence varies from 0.5 to 5% in the literature. The duration of the procedure and repeat procedures are considered to be predisposing factors. The main cause of these infections is thought to be local contamination during the implantation. The commonest causal organism is staphylococcus. Because the presentation may be atypical, the diagnosis is often confirmed by transesophageal echocardiography which is the investigation of choice for imaging a vegetation on an endocavitary pacing lead in cases of infectious endocarditis. The seriousness of this infection requires early diagnosis and adapted treatment including double bactericidal antibiotherapy and complete ablation of the material. Systematic preoperative prophylactic antibiotic therapy is recommended. PMID- 12632763 TI - [Rapid detection of BRCA-1 germline mutations by the protein truncation test in Tunisian families]. AB - Early-onset breast cancer characterize genetic predisposition to cancer in women. BRCA-1 gene was identified as one of the predisposition genes of breast/ovarian cancer. About 90% of the reported mutations in the hereditary breast and ovarian cancer gene, BRCA-1, result in truncated proteins. The aim of our study is to detect rapidly BRCA-1 mutations by the protein truncation test (PTT) in Tunisian women with early breast cancer. Population and methods. We underwent molecular analysis in families with more than: (a) a women under 40 years-old with breast cancer, uni or bilateral; (b) two 1st degree relatives women under 50 years-old with beast cancer. Sixteen women from 12 families were studied by PTT to screen mutations in exon 11 which encodes 61% of BRCA-1. RESULTS: PTT analysis of exon 11 revealed a normal and truncated protein in one patient between 16 from 12 families. CONCLUSION: BRCA-1 gene seems to contribute to at least 1/16 or 6.25% in women with hereditary predisposition to breast/ovarian cancer in Tunisia. PTT promises to be a valuable technique in detecting BRCA-1 mutations in our country. PMID- 12632764 TI - [Bone lesions in multiple myeloma: variation and value of radiologic classification in prognosis]. AB - In this report, we studied the frequency, the types and the prognosis value of the Durie and Salmon's classification of radiological bone lesions in multiple mycloma. Our study concerned 52 patients presenting multiple myeloma, defined according to South West Oncology Group criteria, collected during nine years (1988-1996). Radiological anomalies were noted in 89% of the cases. When we compared the medial survival and the 5 years survival rate in the different groups, we found a statistically significant difference with a better survival for groups 0,1,2 considered all together as compared to group 3 patients (p = 0.0155), (71% versus 37%). According to our series, prognostic value is obviously significant when comparing groups 0, 1, and 2 to group 3. PMID- 12632765 TI - [Ventilatory mechanics and alveolo-capillary diffusion in diabetes]. AB - In order to appreciate the repercussion of diabetes on the respiratory function, we measured the pulmonary volumes, the ventilatory flows, the airways resistances (Raw) and the diffusing capacity for the carbon monoxide (DLCO) of 49 diabetes distributed into 27 IDDM and 22 NIDDM aged from 15 to 56 years, compared to 31 control subjects. We found a significant decrease in the total pulmonary capacity (TCL), the vital capacity (VC), the inspiratory capacity (IC), the ventilatory flows and the DLCO. The decrease of the VC and the FEV1 will be more marked in the IDDM. The decrease of the DLCO will be more pronounced within masculine sex, it seems to be correlated with the duration of diabetes and more impaired if a diabetic nephropathy is associated especially in IDDM. Our results suggest that in the diabetes evolution, the lung is among the target organs in the degenerative complications. The respiratory tests reflect the pulmonary reach in the diabetes and provides important perspectives in the following. PMID- 12632766 TI - [Prognostic impact of karyotype in de novo acute myeloid leukemia: study of 139 cases]. AB - A group of 139 patients with de novo acute myeloid leukemia were investigated to determine the prognostic significance of karyotype on early death, complete remission, continuous complete remission and survival. There were 27 children and 112 adults. Mean age was 32 years. t(15;17) was found associated with a high rate of early death and a diploid karyotype with long continuous complete remission. The presence of a structural change was predictive of shorter survivals. The study of the prognostic impact of recurrent anomalies reveals a good prognostic impact for normal karyotype (1 year survival probability: 40%), followed by t(8;21) (1 year survival probability: 24%), and by t(15;17) (1 year survival probability: 9%). PMID- 12632767 TI - [Predictive elements of malignancy of isolated thyroid nodules]. AB - The clinically revealed thyroid nodules are very frequent (3 to 5% of the general population), among them, fewer than 10% are malignant. The objective of this retrospective study type witness-case is to determinate the malignity predictive clinical and paraclinical elements for 45 patients presenting a thyroid malignant nodule and 45 others matched by age and sex presenting a thyroid isolated nodule. These patients were treated at the ORL ward of Charles Nicolle Hospital over a period of 10 years from 1989 to 1999. The average age of advent of malignant nodule is 43.2 + 14.4 years old with a clear female predominance (sex ratio = 4). The elements clearly linked to the hazard of thyroid cancer are loss of weight (p = 0.05; OR = 8.11), hard consistency of nodule (p = 0.002), the presence of cervical adenopathies (p = 0.05; OR = 4.22), vocal cords paralysis (p = 0.04), the presence of anemia (p = 0.01; OR = 5.7), a solid structure at sonography (p = 0.05) and the presence of a flux at doppler-sonography (p = 0.02). Personal antecedents of non malignant thyroid pathology, the nodule location and size and hypoechogenic features have not been clearly associated with the hazards of thyroid cancer. PMID- 12632768 TI - [Obesity and life style in a population of male school children aged 6 to 10 years in Ariana (Tunisia)]. AB - The increase of the prevalence of the obesity in childhood puts in reason some factors of the environment and the way of life of the child that encourage the hold of weight at these topics of as much more that if they are exposed genetically. A prospective survey has been done close to 3148 school boy aged of 6 to 10 years and who were schooled in the gouvernorat of Ariana showed that the prevalence of the obesity is around 3.7%. A survey case/witness has been achieved in a second time on the way of life of the group of the obese matched to a group of children no obese. An intended questionnaire to parents of children of these 2 groups permitted to collect some informations concerning the weight and the present size of parents, habits of life of their children. The weight to the birth has been searched for in the school medical file or from the notebook of health of the child. The obesity of parents is one factor of risk of the child obesity. The short length of sleep (< 8 hours), the erosion between meals especially in the evening after the dinner, the daily consumption of sugary foods and sparkling drinks is the important risk factors exposing to the infantile obesity this group of age. These behaviours can be corrected by a strategy of prevention and nutritional education. PMID- 12632769 TI - [Typology of Tunisian medical research indexed in Medline from 1965 to 1999]. AB - The medical research is a criteria of a country development and the performance of its health system. This study tried to describe the typology of the Tunisian medical research (themes, journals, types of publication,..). It was a bibliometric and exhaustive survey of Tunisian manuscripts indexed in Medline from 1965 to 1999. Among the 3673 eligible references to the study, 73% have been produced in clinic sciences. The main subjects were: Cancer, hydratic cyst and tuberculosis represented respectively 3.9%, 2.9% and 1.8% of manuscripts. 91% of articles have been published in French. 65% of articles have been published in national journals. This work showed that the medical scientific production remains globally lower in relation to the national human resources, that its social relevance is insufficient in relation to the load of morbidity and that the radiance of the Tunisian medical research would be limited by the choice of no English-writing journals. Thus, the promotion of the national medical research requires the backing of the teaching of: documentary techniques, English and written communication. PMID- 12632771 TI - [Ligation of oropharyngeal mucosa: a rare complication of ligation of esophageal varices. Two case reports]. AB - The eradication of oesophageal varices by binding became the best traitment for the bleeding by rupture of the verices. She tends to substitute the sclerosis because of her fast delay of eradication and her fewer complications and gravity. The incidence of complications after the binding vary between 3.3 and 11%. The most frequent complication is the appearance of oesophageal ulcers that sometimes causes bleeding. A dysphagia can happen within a few hours after the binding. Infections are rare. We report two cases of pharyngeal mucosae binding occurring during the binding of the varices. PMID- 12632770 TI - [Congenital auriculo-ventricular block in the adult]. AB - In this work we report a consecutive series of ten patients having auriculoventricular block "presumed" congenital which is seen in adulthood between 1990 and 2001 to determine their clinical profile and forecast, and to deduct the therapeutic consequences. Our criteria of inclusion requires the existence of patients with a second or third degree heart block, who are less than 40 years old at the time of diagnosis, which is not totally regressive in the effort test and the atropine injection, and whose congenital origin was strongly suspected because of the notion of slow pulse during their youth and the absence of acquired affect which enables us to explain this disease. The population contains ten patients whose average age in hospitalization is 23 years old with the range of 11 to 39, while the average age of the discovery of the disease was 20 years old with the range of 8 to 34. Our patients were referred to us because of cardiac symptoms such as syncope or an equivalent, effort intolerance, asthenia, thoracic pain (like angina), or palpitations. The clinical exam, the electrocardiogram, and additional exams (holter, effort test, echocardiography, electrophysiological investigation) allowed us to retain six indications for the definitive cardiac stimulation, associating to different degrees the existence of worrying symptoms such as a syncope, a congenital heart disorder, a low heart frequency, the association in a sinus dysfunction. Short term and long-term evolution is favorable for patients of the stimulated group as well as the non-stimulated group. PMID- 12632772 TI - [Cyclosporine A in the treatment of autoimmune hepatitis in the child: a case report]. AB - The current immunosuppressive treatment of patients with autoimmune hepatitis consists of prednisone and azathioprine. High doses of prednisone used to obtain the remission of the disease are associated with serious adverse effects. To avoid harmful consequences of prednisone, we proposed to treat patients with oral cyclosporine to obtain the remission of the inflammatory process. CONCLUSION: This case and review of the cases in the literature show that cyclosporine can induce remission of liver disease in children with autoimmune hepatitis, with few and well tolerated adverse effects. PMID- 12632774 TI - U.S. congressman calls for OIG investigation of Joint Commission. PMID- 12632773 TI - [Linguinal metastasis from clear cell carcinoma of the kidney]. AB - Clear cell carcinoma is the most frequent cancer of the kidney (60-80% of cases). This tumour has a great propensity to metastasis. Metastasis lesions appear 15 to 20 years after nephrectomy and differential diagnosis with can be difficult to establish. We report an exceptional case of renal cell carcinoma metastasis to the tongue in a 63-years-old male patient who had a nephrectomy 20 years previously. To our knowledge, this is the second case reported in the literature. PMID- 12632776 TI - Diabetes QI program cuts complications by 60%. PMID- 12632775 TI - JCAHO issues alert on nosocomial infections. PMID- 12632777 TI - Patient Satisfaction Planner. Employee-centered focus sets this system apart. PMID- 12632778 TI - Patient Satisfaction Planner. EDs report success with service guarantees. PMID- 12632779 TI - JCAHO, NCQA form human research quality group. PMID- 12632780 TI - Prevent communication breakdowns. Errors can occur during information transfer. PMID- 12632782 TI - Neonatal group B streptococcus infection in Taiwan: an increasing trend. PMID- 12632781 TI - A boy with hypertension and virilization. PMID- 12632783 TI - In vivo microdialysis study of excitatory and inhibitory amino acid levels in the hippocampus following penicillin-induced seizures in mature rats. AB - To investigate the basic mechanism of epileptogenecity in children, we undertook a study of application of penicillin (PC) to the brain of an animal model. Different doses of PC were injected into the hippocampus of Spraque-Dawley rats subjected to microdialysis to observe in vivo changes of excitatory amino acid (EAA) and inhibitory amino acid (IAA) levels during seizure. All of the EAA and IAA levels increased in the interstitial cerebrospinal fluid (CSF) after focal application of PC. We found that 668 and 1336 units of PC were enough to increase glutamate levels significantly. These levels consistently declined to nearly normal within 40 minutes after PC injection. The aspartate level increased soon after PC injection without statistical significance and then declined almost to baseline level. The IAA, r-aminobutyric acid (GABA), glycine, and taurine levels increased significantly after injection of 1336 u of PC. EEG recording of spike discharges was also well defined after the injection of 1336 u of PC. Our study suggests that an adequate PC dose to the hippocampus will enhance the release of EAA and IAA in the brain as well as epileptogenecity on EEG activity. Using in vivo microdialysis, EAA and IAA can be studied during an induced epileptic process. Elevation of EAA levels soon after focal chemical stimulation of the brain provides a good model for study of epileptogenecity. A delay in the decline of IAA levels may suggest an important phenomenon in seizure suppression and is also worthy of further study. PMID- 12632784 TI - Risk factors of extubation failure in extremely low birth weight infants: a five year retrospective analysis. AB - Extubation failure is one of the most serious complications in extremely low birth weight infants (ELBWI) on mechanical ventilation therapy. We performed a 5 year retrospective analysis to realize the status of extubation failure in ELBWI. Extubation failure was defined as requirements of re-intubation within 72 hours after extubation. The extubation failure rate was 21% (29/138). The mean birth body weight was 808.3 +/- 140.4 gm. The mean gestational age was 25.8 +/- 1.2 wks. The incidence of chronic lung disease (CLD) in infants with extubation failure was 100% (29/29). Apnea of prematurity 49% (14/29) and post-extubation atelectasis 39% (11/29) were the most common reasons for reintubation. The major microbiology findings which correlated with nosocomial pneumonia in infants with extubation failure were Acinetobacter baumanni (21%), Klebsiella pneumonia (21%), Pseudomonas aeroginosa (14%), and Methicillin resistant staphylococcus aureus (14%). In conclusion, post-extubation atelectasis and apnea were the most common reasons for reintubation. ELBWI with extubation failure had higher incidences of post-extubation atelectasis, CLD, and nosocomial pneumonia. Further prospective studies are needed in order to clarify the appropriate extubation program for ELBWI and to prevent post-extubation atelectasis and nosocomial pneumonia. PMID- 12632785 TI - Group B streptococcus infection in infancy: 21-year experience. AB - In this hospital-based review, clinical presentations associated with Group B streptococcus (GBS) infections in children occurring between January 1980 and March 2000 were analyzed. Among the 25 infants with invasive group B streptococcal infections, 9 (36%) were early onset diseases (EOD), 12 (48%) were late onset diseases (LOD), and 4 (16%) occurred beyond the third month of life. Eight of the nine (89%) EOD cases manifested during the first day of life and three (33%) were premature births. Common presentations in GBS infection were fever (75%), poor activity (25%), respiratory distress (25%), lethargy (20%), and irritability (20%). Seizure occurred in 31% of infants with meningitis. Pneumonia (66%) and case-fatality rate (33.3%) were significantly higher in EOD than in LOD. Meningitis was the major manifestation (77%) of LOD and had severe sequelae in 40% of cases. Eight strains were assayed for antibiotic sensitivity and they were all susceptible to penicillin, ampicillin, cefotaxime, and vancomycin. The susceptibilities to erythromycin and clindamycin were 62% and 75%, respectively. Most of the strains from blood or cerebrospinal fluid were type III. PMID- 12632786 TI - Screening of mitochondrial DNA mutations in subjects with non-syndromic familial hearing impairment in Taiwan. AB - BACKGROUND AND PURPOSE: Mitochondrial DNA (mtDNA) mutation is an important cause of hearing impairment, but its prevalence in Taiwan is not clear. The purpose of this study is to identify mtDNA mutations in subjects with non-syndromic familial hearing impairment in Taiwan. METHODS: During a period of 36 months, 10 families with non-syndromic hearing impairment were enrolled. Screening of mutations in the mtDNA, 12S rRNA, and tRNA(Ser) genes were carried out by polymerase chain reaction, restriction enzyme digestion, and direct sequencing. RESULTS: Among the 10 families examined, four were found to have 1555A-->G mutation in the 12S rRNA gene, one had 7511T-->C mutation in the tRNA(Ser) gene, and one had 7444G-->A mutation of mtDNA. CONCLUSION: Up to 60% of families had mtDNA mutations in this study. Therefore, mtDNA mutation seems to be an important cause of non-syndromic familial hearing impairment in Taiwan. PMID- 12632788 TI - Clinical evaluation of short children referred by school screening: an analysis of 655 children. AB - A national screening and referral program of short school children was launched in Taiwan in 2000. We analyzed 655 referrals (boys 303, girls 352) from this program whose heights were below--2 standard deviation score (SDS) for age and gender. Evaluation included: detailed medical history, physical examination and laboratory tests such as blood count, thyroid function, growth hormone screening, bone age and chromosome tests. The results were compared with worldwide data. Normal variations accounted for 64.9% of all etiologies for reasons such as constitutional delay 37.4%, familial short stature 16.5%, and a combination of above two 11.0%. The rest were of pathological short stature for reasons such as: idiopathic short stature 7.9%, growth hormone deficiency 7.9%, precocity 3.2%, skeletal dysplasia 2.3%, intrauterine growth retardation 1.4%, Turner syndrome 1.4%, other chromosomal anomaly 0.8%, and others 5.0%. We conclude that the majority of short stature in Taiwanese children is due to normal variation although potentially treatable causes account for at least 12.8% of cases, such as GHD, Turner syndrome, hypothyroidism and precocity. The inexpensive screening program therefore seems to be beneficial in identifying children with short stature of potentially treatable etiology. PMID- 12632787 TI - Failure to thrive in children with primary distal type renal tubular acidosis. AB - Primary distal type renal tubular acidosis (RTA-type1) results from defects of distal renal tubules in urinary acidification. In attempt to evaluate the clinical features, growth and outcome of primary distal type RTA in Taiwan, we retrospectively studied 28 patients (16 males and 12 females) of primary distal type RTA in our hospital in the past 13 years. The mean age at diagnosis was 2 years and 6.8 months old. Hematuria was found in 5 out of 25 cases (20%). Nephrocalcinosis was found in 5 out of 21 cases (23.8%). The mean value of Uca/Ucr was 0.313 +/- 0.067 in those older than 2 years and 0.262 +/- 0.152 in those younger than 2 years. Rickets was suspected in only one child by radiologic study. At initial diagnosis, the mean bone age was delayed for 16 months. The older the patient was, the more delayed was the bone age. The mean value of height was -2.81 standard deviation score (SDS); the mean value of body weight was -2.44 SDS; and the mean value of growth velocity in height was -2.62 SDS. After alkali therapy given from 8.5 months to 5 years and 7 months, improvement was found in height up to -1.69SDS, in body weight up to -1.10SDS, and in height velocity up to 1.06SDS. PMID- 12632789 TI - Spontaneous spinal epidural hematoma in a 5-year-old girl. AB - Spontaneous spinal epidural hematoma (SSEH) is rare in children and is usually confined to the dorsal epidural space. We report on a 5-year-old girl who suffered from acute paraplegia and urinary retention. Magnetic resonance imaging of the spine revealed a dorsal epidural mass from the level of T6 to T8. Emergent decompressive laminectomy was performed and the epidural hematoma was evacuated. She received rehabilitation after surgery with complete recovery 2 months later. PMID- 12632790 TI - Esophageal atresia associated with tracheal stenosis and right lung agenesis: report of one case. AB - Unilateral pulmonary agenesis with esophageal atresia is a rare condition. Patients with this condition almost die of respiratory failure during early infancy. We describe a case of a premature female infant who has tracheal stenosis, right lung agenesis and esophageal atresia at the same time. Because of respiratory failure, this baby passed away at the age of 39 days. PMID- 12632791 TI - Intrapulmonary thrombolytic therapy in a child with acute pulmonary embolism due to primary antiphospholipid syndrome. AB - Pulmonary embolism (PE) is a rare but life-threatening condition, which needs emergent treatment in children. Though anticoagulation is the standard treatment for PE, recent studies revealed intrapulmonary thrombolytic therapy may produce more rapid clot lysis and less systemic complications in adult patients. We reported a 10-year-old girl who had diffused multiple pulmonary emboli due to primary antiphospholipid syndrome. The thrombi persisted in spite of intravenous heparin infusion for 3 days, which resolved quickly after intrapulmonary streptokinase infusion for 12 hours. PMID- 12632792 TI - Fibrous dysplasia of mandible with chronic osteomyelitis in a child: report of one case. AB - We report here a 6-year-old girl with fibrous dysplasia (FD) of the mandibular bone. She had a growing mass with local pain over right chin after a severe trauma, which was thought to be chronic osteomyelitis (OM). After failure of antibiotic treatment, malignant bone tumor was suspected from imaging studies including magnetic resonance imaging (MRI). FD of the mandible with chronic OM was confirmed one year after its onset by repeated biopsy performed at our hospital. Causes of delayed diagnosis may include (1) FD of the mandibular bone and chronic OM have similar characteristics clinically and radiographically, (2) the previous biopsy was not performed at appropriate site, and (3) failure to include fibrous dysplasia in the differential diagnosis. In this report, we also review the features in radiology and MRI of OM and FD, which may help differentiate the diagnosis. When a patient with mandibular FD has acute symptoms, the possibility of superimposed OM or malignant change should be considered. PMID- 12632793 TI - Relapsing hepatitis A in children: report of two cases. AB - Viral A hepatitis is a self-limited infection occurring predominantly among children usually as an anicteric often subclinical illness. After a stage of typical hepatitis A, a biphasic or relapsing form of viral hepatitis A (R-HA) may occur. Although relapse occurs in 3 to 21% of patients with acute hepatitis A, the patients with R-HA have been unusually reported in the literature. In this article, we report two children with R-HA because of unusual presentation and to draw attention to the importance of R-EA. PMID- 12632794 TI - Invasive pneumococcal infections among Canadian aboriginal children. PMID- 12632795 TI - National Advisory Committee on Immunization (NACI). Statement on thimerosal. PMID- 12632796 TI - [Role of transcranial magnetic stimulation in clinical diagnosis: facial nerve neurography]. AB - Facial nerve neurography involving magnetic stimulation techniques can be used to assess the intracranial segment of the facial nerve and the entire facial motor pathway, as opposed to the traditional neurography, involving only extracranial electric stimulation of the nerve. Both our own experience and data published in the literature underline the value of the method in localising facial nerve dysfunction and its role in clinical diagnosis. It is non-invasive and easy to perform. Canalicular hypoexcitability has proved to be the most useful and sensitive parameter, which indicates the dysfunction of the nerve between the brain stem and the facial canal. This is an electrophysiological finding which offers for the first time positive criteria for the diagnosis of Bell's palsy. The absence of canalicular hypoexcitability practically excludes the possibility of Bell's palsy. The technique is also able to demonstrate subclinical dysfunction of the nerve, which can be of considerable help in the etiological diagnosis of facial palsies. For example, in a situation where clinically unilateral facial weakness is observed, but facial nerve neurography demonstrates bilateral involvement, etiologies other than Bell's palsy are more likely, such as Lyme's disease, Guillain-Barre syndrome, meningeal affections etc. Furthermore, the technique differentiates reliably between peripheral facial nerve lesion involving the segment in the brain stem or the segment after leaving the brainstem. PMID- 12632797 TI - [Effectiveness of autologous free fat graft and of Spongostan in preventing scar formation after microdiscectomy]. AB - INTRODUCTION: Postoperative scar formation has been implicated as one of the possible cause of persistent or recurrent pain after spine surgery. The efficacy of the autologous free fat graft and the Spongostan gel foam for the prevention of extradural adhesion after lumbar discectomy was investigated by the evaluation of postoperative neurological symptoms and visual analogue scale. PATIENTS AND METHODS: In the study 174 patients were involved, average 14.5 months after the intervention. Autologous free fat graft (group I) and Spongostan gel foam (group II) was used randomly by the operations. Patients were operated by the same surgeon, and they were investigated by another independent surgeon. RESULT: No significant difference was found between the two groups. PMID- 12632798 TI - [Correlation between the effectiveness of atypical antipsychotics and dopamine D3 receptor polymorphism in schizophrenia]. AB - OBJECT: Numerous relevant variants of dopamine receptors have been identified in schizophrenia. The Ser9Gly gene polymorphism of dopamine D3 receptor is known as a susceptibility factor for the disease. In addition, it has a role in the modification of therapeutic effect of antipsychotics. In this naturalistic study the authors investigated the relationship between this polymorphism and the therapeutic response to atypical antipsychotics. METHOD: 75 patients with schizophrenia according to DSM-IV and 45 healthy controls were recruited. The patients were divided to responder and nonresponder subgroups, cut-off: > 20 point improvement in Global Assessment of Functioning. By polymerase chain reaction the genotype of dopamine D3 receptor of every participant was determined. RESULTS: The Ser9Ser genotype of dopamine D3 receptor was more frequent in the nonresponder subgroup (64%, p = 0.0018). The Ser9 allele was overrepresented among nonresponder patients (82%, p = 0.0172). CONCLUSION: Based on our results, the worse therapeutic response to atypical antipsychotics is associated with Ser9 variant of dopamine D3 receptor. PMID- 12632799 TI - [Friedreich ataxia--diagnosis after a decade. Differential diagnosis of inherited spinocerebellar ataxias]. AB - The clinical diagnosis of inherited spinocerebellar ataxias is difficult, because phenotypes frequently overlap. The authors attempt to review the different inherited ataxia syndromes, discussing the most frequent one, Friedreich-ataxia in detail. The case of a patient with Friedreich-ataxia is presented, where the genetically supported diagnosis has been made more than ten years following the onset of the symptoms, after several hospitalizations and misdiagnosis. The correct diagnosis can be established based on the Geoffroy-Harding criteria and gene mutation analysis. PMID- 12632801 TI - [Utility of electroshock therapy in epilepsy-associated psychosis]. AB - Psychiatric disorders which indicate the use of electroconvulsive therapy (ECT) also occur in epileptic patients, but there is a lack of medical authority concerning the use of ECT in epileptic patients. This is surprising because in recent years it has been proved that ECT has an anti-convulsive effect to some degree. A case study of an epileptic patient is presented whose progress has been monitored for several years. Antiepileptic drugs were seemingly able to control his epilepsy but at the same time progressive behavioural disturbance (schizophreniform psychosis) accompanied by agitation and violent behaviour developed. Considering the recurrent psychotic decompensations and the relative ineffectiveness of antipsychotics, the authors decided to administer ECT. As a result they were able to bring about the longest symptom free balanced period in the patient. According to the data based on previous medical studies and the experience they can suppose that ECT is not immediately contraindicated by the presence of epilepsy with active interictal focus if the psychopathological condition makes it necessary. In view of the epileptogenic risk factors of classical neuroleptics, the epileptogenic effects of accompanying psychosis and the probable anticonvulsive potential of ECT in cases of severe psychiatric complications accompanying epilepsy ECT could be used more frequently. PMID- 12632800 TI - [Hormone disorders caused by intrasellar persistent trigeminal artery]. AB - Persistent trigeminal artery is a relatively frequent type of intracranial arterial developmental anomalies. The diagnostic tools for demonstration previously consisted of carotid angiography, later CT and DSA and nowadays MR and MRA. The practical benefit of the diagnosis is to avoid any hazard at the operation of associated hypophysis adenomas and aneurysms and could also give a possible explanation for apparent hormonal abnormalities. The authors support the latter possibility with a case report. PMID- 12632803 TI - What becomes an icon most? AB - Some brands become icons. Think of Nike, Apple, Harley-Davidson: They're the brands every marketer regards with awe. But they are not built according to the principles of conventional marketing, says Harvard Business School marketing professor Douglas Holt. Iconic brands beat the competition not just by delivering innovative benefits, services, or technologies but by forging a deep connection with the culture. A brand becomes an icon when it offers a compelling myth, a story that can help people resolve tensions in their lives. The deepest source of tension in modern society is the disparity between national ideology and the average citizen's reality. When ideologies shift, myths become even more important, and in America, the most potent myths are depictions of rebels. Mountain Dew has long offered a rebel myth in ads showing exciting, vital men who are far from the ideological model of success. Loyal customers drink the beverage to consume the myth. But Mountain Dew's greatest achievement is that it has retained its iconic power by creating fresh rebel myths to suit the tensions of each era: first the hillbilly, who stood in stark contrast to the organization man of the 1950s and 1960s; then the redneck, who rebelled against the investment bankers and consultants of the 1970s and 1980s; and most recently the slacker, who rejects the values and behaviors that, for the past decade, have marked the successful executive. Holt says marketers can learn from Mountain Dew and other iconic brands if they are willing to move beyond conventional brand management and acquire knowledge and skills they may not have. They must learn to target national contradictions instead of just consumer segments, create myths that make sense of confusing societal changes, and speak with a rebel's voice. PMID- 12632802 TI - Recent advances in Parkinson's disease. PMID- 12632804 TI - Bottom-feeding for blockbuster businesses. AB - Marketing experts tell companies to analyze their customer portfolios and weed out buyer segments that don't generate attractive returns. Loyalty experts stress the need to aim retention programs at "good" customers--profitable ones- and encourage the "bad" ones to buy from competitors. And customer-relationship management software provides ever more sophisticated ways to identify and eliminate poorly performing customers. On the surface, the movement to banish unprofitable customers seems reasonable. But writing off a customer relationship simply because it is currently unprofitable is at best rash and at worst counterproductive. Executives shouldn't be asking themselves, How can we shun unprofitable customers? They need to ask, How can we make money off the customers that everyone else is shunning? When you look at apparently unattractive segments through this lens, you often see opportunities to serve those segments in ways that fundamentally change customer economics. Consider Paychex, a payroll processing company that built a nearly billion-dollar business by serving small companies. Established players had ignored these customers on the assumption that small companies couldn't afford the service. When founder Tom Golisano couldn't convince his bosses at Electronic Accounting Systems that they were missing a major opportunity, he started a company that now serves 390,000 U.S. customers, each employing around 14 people. In this article, the authors look closely at bottom-feeders--companies that assessed the needs of supposedly unattractive customers and redesigned their business models to turn a profit by fulfilling those needs. And they offer lessons other executives can use to do the same. PMID- 12632805 TI - For the last time: stock options are an expense. AB - Should stock options be recorded as an expense on a company's income statement and balance sheet, or should they remain where they are, relegated to footnotes? The extraordinary boom in share prices during the Internet bubble made critics of option expensing look like spoilsports. But since the crash, the debate has returned with a vengeance. And no wonder: The authors believe the case for expensing options is overwhelming. In this article, Nobel Iaureate Robert Merton, one of the inventors of the Black-Scholes option-pricing model; his coauthor on the classic textbook Finance, Zvi Bodie; and Robert Kaplan, creator of the Balanced Scorecard, examine and dismiss the principal claims put forward by those who continue to oppose options expensing. They demonstrate that stock-option grants do indeed have real cash-flow implications that need to be reported. They show that effective ways certainly exist to quantify those implications. They detail the distortions that relegating stock-option accounting to footnotes creates. And they show why reporting option costs should in no way hamper young companies in their efforts to provide incentives. Options are indeed a powerful incentive, the authors agree, and failing to record a transaction that creates such powerful effects is economically indefensible. Worse, it encourages companies to favor options over alternative incentive systems. It is not the proper role of accounting standards, the authors argue, to distort executive and employee compensation by subsidizing one particular form of compensation and no other. Companies should choose compensation methods according to their economic benefits--not the way they are reported. PMID- 12632806 TI - Predictable surprises: the disasters you should have seen coming. AB - Think hard about the problems in your organization or about potential upheavals in the markets in which you operate. Could some of those problems--ones no one is attending to--turn into disasters? If you're like most executives, you'll sheepishly answer yes. As Harvard Business School professors Michael Watkins and Max Bazerman illustrate in this timely article, most of the "unexpected" events that buffet companies should have been anticipated--they're "predictable surprises." Such disasters take many forms, from financial scandals to disruptions in operations, from organizational upheavals to product failures. Some result in short-term losses or distractions, while others cause damage that takes years to repair. Some are truly catastrophic--the events of September 11, 2001, are a tragic example of a predictable surprise. The bad news is that all companies, including your own, are vulnerable to predictable surprises. The good news is that recent research helps explain why that's so and what companies can do to minimize their risk. The authors contend that organizations' inability to prepare for predictable surprises can be traced to three sets of vulnerabilities: psychological, organizational, and political. To address these vulnerabilities, the authors recommend the RPM approach. More than just the usual environmental scanning and contingency planning, RPM requires a chain of actions--recognizing, prioritizing, and mobilizing--that companies must meticulously adhere to. Failure to apply any one of these steps, the authors say, can leave an organization vulnerable. Given the extraordinarily high stakes involved, it should be every business leader's core responsibility to apply the RPM approach, the authors conclude. PMID- 12632807 TI - The Board's missing link. AB - If a dam springs several leaks, there are various ways to respond. One could assiduously plug the holes, for instance. Or one could correct the underlying weaknesses, a more sensible approach. When it comes to corporate governance, for too long we have relied on the first approach. But the causes of many governance problems lie well below the surface--specifically, in critical relationships that are not structured to support the players involved. In other words, the very foundation of the system is flawed. And unless we correct the structural problems, surface changes are unlikely to have a lasting impact. When shareholders, management, and the board of directors work together as a system, they provide a powerful set of checks and balances. But the relationship between shareholders and directors is fraught with weaknesses, undermining the entire system's equilibrium. As the authors explain, the exchange of information between these two players is poor. Directors, though elected by shareholders to serve as their agents, aren't individually accountable to the investors. And shareholders- for a variety of reasons--have failed to exert much influence over boards. In the end, directors are left with the Herculean task of faithfully representing shareholders whose preferences are unclear, and shareholders have little say about who represents them and few mechanisms through which to create change. The authors suggest several ways to improve the relationship between shareholders and directors: Increase board accountability by recording individual directors' votes on key corporate resolutions; separate the positions of chairman and CEO; reinvigorate shareholders; and give boards funding to pay for outside experts who can provide perspective on crucial issues. PMID- 12632809 TI - Personalize your management development. AB - Most organizations struggle with leadership development. They promote tope performers into management roles, put them through a few workshops and seminars, then throw them to the wolves. Managers with the ability to survive and thrive are rewarded; those without it are disciplined or reassigned. The problem is, an alarming number of people fall into the second category. This happens not because managers lack skills but because companies fail to realize that there is no single kind of leader-in-training. In this article, Natalie Shope Griffin, a consultant in executive and organizational development at Nationwide Financial, describes four kinds of manager-in-training, each embodying unique challenges and opportunities. Reluctant leaders appear to have all the necessary skills to be excellent managers but can't imagine themselves succeeding in a leadership role. Arrogant leaders have the opposite problem; they believe they already possess all the management skills they'll ever need. Unknown leaders are overlooked because they don't develop relationships outside of a small circle of close colleagues. Finally, there are the workaholics who put work above all else and spend 100 hours a week in the office. The author outlines specific training approaches tailored to each type of prospective leader. By focusing on the unique circumstances of individual managers, investing in them early in their careers, offering effective coaching, and providing real-life management experiences, Nationwide's leadership-development program has produced hundreds of successful leaders. PMID- 12632808 TI - Why hierarchies thrive. AB - Hardly anyone has a good word to say about hierarchies. Academics, consultants, and management gurus regularly forecast their imminent replacement because hierarchies--even when populated by considerate and intelligent people--can be cruel and stupid. They routinely transform motivated and loyal employees into disaffected Dilberts. It's no wonder that we continue to search for more humane and productive alternatives to them. Yet the intensity with which we struggle against hierarchies only serves to highlight their durability. Hierarchy, it seems, may be intrinsic not only to the natural world but also to our own natures. In this article, organizational behavior expert Harold J. Leavitt presents neither a defense of human hierarchies nor another attack on them. Instead, he offers a reality check, a reminder that hierarchy remains the basic structure of most, if not all, large, ongoing human organizations. That's because although they are often depicted as being out of date, hierarchies have proved to be extraordinarily adaptive. Over the past 50 years, for example, they have co opted the three major managerial movements--human relations, analytic management, and communities of practice. Hierarchies also persist because they deliver real practical and psychological value, and they fulfill our deep need for order and security. Despite the good they may do, hierarchies are inevitably authoritarian. That authoritarianism shows up in all kinds of ways and influences everything in organizations, particularly communication. In multilevel organizations, for instance, messages get distorted as they travel up and down the ladder of command. Self-protection and self-interest weigh in, and relevant information is lost as messages make stops along the route. Sensitive leaders take steps to make speaking the truth as painless as possible. But it never is in organizations, because authoritarianism is an immutable element of hierarchy. PMID- 12632810 TI - Finding your innovation sweet spot. AB - Most new product ideas are either uninspired or impractical. So how can developers hit the innovation sweet spot--far enough from existing products to attract real interest but close enough that they are feasible to make and market? They can apply five innovation patterns that manipulate existing components of a product and its immediate environment to come up with something both ingenious and viable, say the authors. The subtraction pattern works by removing product components, particularly those that seem desirable or indispensable. Think of the legless high chair that attaches to the kitchen table. The multiplication pattern makes one or more copies of an existing component, then alters those copies in some important way. For example, the Gillette double-bladed razor features a second blade that cuts whiskers at a slightly different angle. By dividing an existing product into its component parts--the division pattern--you can see something that was an integrated whole in an entirely different light. Think of the modern home stereo--it has modular speakers, tuners, and CD and tape players, which allow users to customize their sound systems. The task unification pattern involves assigning a new task to an existing product element or environmental attribute, thereby unifying two tasks in a single component. An example is the defrosting filament in an automobile windshield that also serves as a radio antenna. Finally, the attribute dependency pattern alters or creates the dependent relationships between a product and its environment. For example, by creating a dependent relationship between lens color and external lighting conditions, eyeglass developers came up with a lens that changes color when exposed to sunlight. PMID- 12632811 TI - How green buildings are smarter and safter. PMID- 12632812 TI - There are numerous ways to determine just how sustainable so-called green products are today. PMID- 12632813 TI - Determinants of successful surgical revascularization for failed angioplasty in patients with acute myocardial infarction and cardiogenic shock. AB - BACKGROUND AND PURPOSE: Previous studies have reported encouraging results with percutaneous transluminal coronary angioplasty (PTCA) in patients with acute myocardial infarction (AMI) and cardiogenic shock (CS). However, PTCA fails in many patients and the mortality rate associated with the procedure is very high. This study sought to assess the determinants of successful coronary artery bypass grafting (CABG) in patients with AMI, CS and failed PTCA. MATERIALS AND METHODS: From April 1997 to May 2001, 33 patients with a mean age of 62.9 +/- 10.5 years underwent CABG for AMI, CS and failed PTCA. PTCA failed or was not attempted because of primary failure in 13 patients, reocclusion in three, no-reflow in one, and multivessel disease in 18. Patients were initially stabilized with inotropic infusion and mechanical circulatory support. Before June 2000, patients underwent conventional CABG, while after June 2000, patients underwent beating heart CABG. Risk factor analysis was performed using retrospective chart review. RESULTS: The overall in-hospital mortality rate was 30.3%-33.3% in 24 patients who underwent conventional CABG and 22.2% in nine patients who underwent beating heart CABG. Risk factors for death were unstable hemodynamics (p < 0.001), non right coronary artery as the infarct-related artery (IRA) (p = 0.012), high inotropic dose (p = 0.013), intubation (p = 0.021), cardiopulmonary resuscitation (p = 0.021), and early bypass (p = 0.050). CONCLUSIONS: CABG was associated with a high in-hospital mortality rate in patients with AMI, CS and failed PTCA. Patients with unstable hemodynamics and high inotropic dose before surgery had the worst survival rate. PMID- 12632814 TI - Operative strategy in follicular thyroid cancer. AB - BACKGROUND AND PURPOSE: Surgery is the main treatment for most thyroid cancers. However, the indications for subtotal or radical thyroidectomy in cases of follicular thyroid cancer remain controversial. This study examined the indications for subtotal thyroidectomy or radical total thyroidectomy in patients with follicular cancer. METHODS: A total of 196 cases of follicular cancer diagnosed among 1,545 thyroid cancer cases between 1960 and 2000 at National Taiwan University Hospital (NTUH) were included in this study. Diagnosis of follicular cancer was based on the findings of pathologic examination. All surgeries were performed in the Department of Surgery at NTUH. The average period of postoperative follow-up was 11.7 years. Subtotal thyroidectomy was defined as thyroid resection performed on not more than two-thirds of the whole lobe. Radical thyroidectomy was defined as total removal of the thyroid plus cervical lymph node removal if surgical examination revealed malignancy. Local recurrence was defined as tumor reappearance in soft tissue in the neck or cervical lymph node after initial treatment. Distal metastasis was also surveyed from medical records. Patients were classified into three groups according to the size of the surgical specimen as less than 3 cm in diameter, 3 to 5 cm, and more than 5 cm. RESULTS: Between 1960 and 2000, the percentage of follicular cancer patients undergoing follicular cancer surgery decreased from 22.0 to 7.7%, while papillary cancer surgery increased from 65.5 to 87.1%. This change was associated with the addition of iodine to table salt island-wide from 1967. The change in percentage of surgically treated cases was greater between 1991 and 2000. Subtotal thyroidectomy was performed in 56 cases and radical thyroidectomy was performed in 140 cases during the study period. None of the 63 cases with a tumor diameter of less than 3 cm undergoing either radical or subtotal thyroidectomy had local recurrence during a mean follow-up of 11.7 years. However, six of those cases (9.5%) had distant metastasis. CONCLUSIONS: For follicular cancers of less than 3 cm in diameter, subtotal thyroidectomy instead of radical thyroidectomy may be considered, but close follow-up is necessary. PMID- 12632815 TI - Combined effects of chronic hepatitis virus infections and substance-use habits on chronic liver diseases in Taiwanese aborigines. AB - BACKGROUND AND PURPOSE: Chronic liver disease (CLD) is a major cause of death in Taiwanese aborigines. The roles of substance-use habits and hepatitis B virus (HBV) and hepatitis C virus (HCV) infections in the development of CLD are not well understood in this indigenous population. METHODS: A hospital-based, case control study of 79 consecutive CLD patients and 107 non-CLD controls was performed. Serostatus of hepatitis B surface antigen (HBsAg) and antibodies against hepatitis C virus (anti-HCV) were determined in all subjects. Each subject completed an epidemiologic questionnaire about the risk factors of CLD. RESULTS: Atayal ethnicity, alcohol drinking, cigarette smoking, betel quid chewing, seropositivity of HBsAg and anti-HCV antibodies were associated with a significantly elevated CLD risk. In the combinatory analyses of HBsAg serostatus and substance-use habits, HBsAg-positive substance users had the highest CLD risk, followed by HBsAg-positive non-users, HBsAg-negative users, and HBsAg negative non-users. Similarly, anti-HCV-positive alcohol drinkers and betel quid chewers had greater CLD risks than other groups. The multivariate-adjusted odds ratios (ORs) for males, Atayal ethnicity and seropositivity of HBsAg and anti-HCV were significantly elevated. There was a biologic gradient in the risk of developing CLD associated with the number of substance-use habits. The multivariate-adjusted ORs were 4.7 (95% confidence interval [CI], 1.3-16.8) and 7.9 (95% CI, 2.1-30.4) for subjects with 1-2 and 3 habits, respectively, compared to subjects with no habit. CONCLUSION: Our data suggest that chronic HBV and HCV infections, alcohol drinking, cigarette smoking, and betel quid chewing play important roles in the development of CLD in Taiwanese aborigines. PMID- 12632816 TI - Risk factors for intestinal metaplasia in adult residents of Matzu: a cross sectional study. AB - BACKGROUND AND PURPOSE: People living on Matzu, a group of small islets, have the highest mortality rate from gastric cancer (GC) in Taiwan. Intestinal metaplasia (IM) is a precursor lesion of GC. We therefore conducted a gastroscopic screening program in this district to evaluate the risk factors associated with IM. METHODS: A total of 274 residents of Matzu, aged 30 years or older, underwent testing for anti-Helicobacter pylori IgG antibody and serum pepsinogen (PG) I/PG II levels, and gastroscopic examination. Univariate and multivariate logistic regression models were used to identify risk factors associated with IM in terms of odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: Among the 274 subjects, 118 (43%) were confirmed to have IM by histologic examination. Individuals aged 60 years or older had a 2.66 times higher risk (95% CI 1.38 5.13) of having IM, as compared with those less than 60 years of age. Subjects with duodenal ulcer had a 60% lower risk of coexisting IM (OR, 0.40; 95% CI, 0.19 0.83). Meanwhile, subjects with gastric ulcer were not at increased risk of IM (OR, 0.84; 95% CI, 0.34-2.08). Low serum PG I level, low PG I/PG II ratio, and positive anti-H. pylori IgG antibody, were associated with IM in the univariate analysis. While age and duodenal ulcer maintained their independent effects in the multivariate analysis, only a low PG I/PG II ratio and H. pylori positivity were significantly associated with IM. CONCLUSIONS: This study of residents of Matzu found that age and H. pylori infection are risk factors for IM, while subjects with duodenal ulcer have a lower incidence of coexisting IM. Low PG I/PG II ratio is a potential noninvasive biomarker of IM. PMID- 12632818 TI - Functional recovery after hip fracture: six months' follow-up of patients in a multidisciplinary rehabilitation program. AB - BACKGROUND AND PURPOSE: Hip fracture is a major source of disability among the elderly. The purpose of this study was to evaluate the effects of an in-hospital multidisciplinary rehabilitation program (MRP) on basic activities of daily living (BADL) and mobility 6 months after hospital discharge in patients with hip fractures. METHODS: A before and after quasi-experimental design was used. Subjects were recruited in a large teaching hospital in Taipei, Taiwan. The first 44 consecutive patients recruited received conventional care (control group). The next 50 consecutive patients received physical therapy, nursing-supervised practice of exercise, and discharge planning (intervention group). The 94 patients recruited were aged 60 years or older and were hospitalized to receive open reduction and internal fixation or arthroplasty. Subjects were assessed at admission, on the fourth operative day, on the day of discharge, and 6 months after discharge. RESULTS: On average, subjects did not regain their prefracture functional score for BADL and mobility 6 months after discharge. The intervention versus control group had a lower incidence of functional decline in BADL (16.0% vs. 34.1%; p = 0.01) and mobility (48.0% vs. 75.0%; p = 0.01) 6 months after discharge. Subjects who were independent before fracture (odds ratio [OR], 12.24), did not receive MRP intervention (OR, 4.63), or who were female (OR, 5.24), were more likely to have a decline in function 6 months after discharge. CONCLUSION: An MRP had a continuous positive effect on hip-fracture patients, thus facilitating improved recovery in BADL and mobility 6 months after discharge. PMID- 12632817 TI - Screening for pulmonary tuberculosis among military conscripts in Taiwan. AB - BACKGROUND AND PURPOSE: Military service is obligatory for young men who are medically fit in Taiwan. Each year, all 19-year-old men are notified to undergo a required health check to determine their eligibility for military service. This study determined the prevalence of pulmonary tuberculosis (TB) among military conscripts in 1997 and 1998 and evaluated the effect of the mass radiographic screening program for military conscripts on the reported TB case rate. METHODS: During the annual health check, all 19-year-old men in Taiwan undergo miniature (70 x 70 mm) chest roentgenography (CXR). Those who have suspicious lesions on miniature films are notified to undergo follow-up CXR (14 x 14 inch) and sputum examinations. All these data for the period from 1997 to 1998 were analyzed to determine the prevalence of pulmonary TB. To evaluate the effect of the mass radiographic screening program among military conscripts on the reported TB case rates, annual data for newly diagnosed pulmonary TB in 1997 and 1998 were obtained from the National TB Register and analyzed by age and sex. RESULTS: A total of 305, 140 men eligible for military service underwent the required examination in 1997 and 1998. Pulmonary TB was diagnosed in 237 (0.08%) of these men, and 21 (6.9/100,000) had positive bacteriologic findings. In 1997 and 1998, the annual reported TB case rates for males and females were similar in both children and young adults, except for an early peak among men aged 19 years. CONCLUSION: The screening program resulted in a significant increase in the reported TB case rate among males aged 19 years. However, a low bacteriologic confirmation rate implies the possibility of over-diagnosis with CXR. Mass radiographic screening at the health check for military conscripts is inefficient in detecting bacteriologically confirmed TB, an observation with implications for screening policy development by the national TB program. PMID- 12632819 TI - Adrenal lymphoma and Addison's disease: report of a case. AB - Addison's disease due to adrenal lymphoma usually manifests as bilateral adrenal enlargement. We report a patient with Addsion's disease in whom the initial overt primary adrenal insufficiency was accompanied by an only slightly enlarged right adrenal gland. The 80-year-old man presented with nausea, anorexia, weight loss, and hyperpigmentation of the skin and buccal mucosa. Addison's disease was diagnosed based on this clinical presentation and laboratory findings of low cortisol and high adrenocorticotropin levels. Computerized tomography (CT) of the adrenal glands revealed a small right adrenal tumor. His family refused to allow percutaneous or surgical biopsy to determine the nature of the tumor. His general condition improved after steroid supplementation. However, about 1 year later, dizziness, fever, night sweats, and edema of the lower legs developed, and adrenal CT scanning revealed that the left adrenal gland had enlarged and now exceeded the size of the right one. Left adrenalectomy was performed and pathology showed a diffuse large B-cell lymphoma. Staging work-up using whole body CT scanning suggested a stage IIIb lymphoma. Chemotherapy was given, but the disease still progressed and the patient died 4 months after diagnosis. Primary adrenal lymphoma should be considered in the differential diagnosis of Addison's disease, even if only slight enlargement of the adrenal glands is found initially. PMID- 12632820 TI - Treatment of thrombotic microangiopathy in pregnancy with plasma exchange: a report of two cases. AB - Thrombotic microangiopathy is a rare disease that can be induced and precipitated by pregnancy, and is associated with high maternal and fetal morbidity and mortality. It results from abnormal intravascular platelet aggregation that leads to transient ischemia in various organs, including the central nervous system, kidneys and placenta. Plasma exchange is the most widely accepted method of treatment for this condition. Delayed diagnosis is the main reason for morbidity and mortality, and results from difficulty in differentiating thrombotic microangiopathy from other obstetric emergencies. We report two cases of thrombotic microangiopathy that occurred antepartum and postpartum, respectively. The first patient was a 33-year-old woman who had two previous episodes of intrauterine fetal death in the 13th and 28th weeks of gestation, respectively. She received early plasma exchange at the 23rd week of gestation during this pregnancy and the fetus was delivered uneventfully. The second patient was a 28 year-old woman with progressive thrombocytopenia, anemia and deterioration of renal and liver function postpartum. She received early plasma exchange and it markedly improved her thrombocytopenia without sequelae. In conclusion, early diagnosis and early initiation of plasmapheresis may improve both maternal and fetal prognosis in thrombotic microangiopathy. PMID- 12632821 TI - Neural-crest tumor presenting with chronic diarrhea: a report of three cases. AB - The association of chronic diarrhea with neural-crest tumors is uncommon. In the past 12 years, we encountered three cases of neural-crest tumors presenting initially as chronic diarrhea. The incidence of chronic diarrhea in patients with neural-crest tumors at our hospital during this period was 3.8%. These patients (two girls and one boy aged between 15 to 28 mo) presented with a 3-week to 6 month history of chronic diarrhea of unknown cause. A posterior mediastinal mass or abdominal mass found on sonography and chest roentgenography eventually led to the diagnosis of neural-crest tumor in these patients. Pathology revealed ganglioneuroblastoma in two cases and neuroblastoma in one. All three patients received chemotherapy and underwent surgery. Diarrhea ceased postoperatively in all three patients. Although one of the patients died of septic shock during chemotherapy, the other two remained free of disease at 30 and 22 months of postoperative follow-up, respectively. A high index of suspicion is needed to identify cases of neural-crest tumor from the presenting symptom of chronic diarrhea. PMID- 12632822 TI - Postpartum hemolytic uremic syndrome following abruptio placenta: report of a case. AB - Postpartum hemolytic uremic syndrome (HUS) is an unusual complication that presents with microangiopathic hemolytic anemia, thrombocytopenia, and acute renal failure after delivery. In this report, we describe a 32-year-old patient (gravida 3, para 1, artificial abortion 1) who developed postpartum HUS following abruptio placenta. After cesarean delivery due to abruptio placenta, the patient developed acute renal failure, microangiopathic hemolytic anemia, and thrombocytopenia followed by hypertension. Plasma exchange led to recovery from thrombocytopenia and improvement in renal function. This case highlights the importance of observation of peripheral blood smears in patients with abruptio placenta who develop thrombocytopenia after delivery. PMID- 12632823 TI - Small-cell lung cancer presenting with Lambert-Eaton myasthenic syndrome and respiratory failure. AB - Lambert-Eaton myasthenic syndrome (LEMS) is a neuromuscular disorder characterized by defective neurotransmitter release at presynaptic terminals. It is caused by an IgG autoantibody reacting against voltage-gated calcium channels. Severe LEMS complicated by ventilatory failure is rare. We report a case of small cell lung cancer (SCLC) presenting with LEMS and ventilatory failure in a 67-year old man who initially presented with progressive limb weakness for 6 months and tachypnea with shallow breathing for 1 week. LEMS was diagnosed through electrophysiologic studies. Chest radiography and computerized tomography showed a huge mass lesion over the left anterior and middle mediastinum with an encasement of the left pulmonary artery. Cytologic examination of ultrasound guided fine needle aspiration disclosed SCLC. Successful treatment in combination with plasma exchange and chemotherapy resulted in dramatic tumor regression and LEMS remission, which were confirmed by chest radiography and electrophysiologic studies. This case suggests that plasma exchange and chemotherapy can be effective in treating SCLC with severe LEMS that produces ventilatory failure. PMID- 12632824 TI - Tobacco isolated as a cause of skin aging still another reason to quit smoking! PMID- 12632825 TI - A descriptive epidemiologic study of HIV-infected individuals in Hawaii: report of the Hawaii Sero-Positivity and Medical Management database (HSPAMM). AB - This is a retrospective study of the HSPAMM database evaluating differences in clinical, laboratory, HIV-risk factors and demographic characteristics with respect to gender and ethnicity. There were no significant differences comparing gender, and Hawaiians and non-Hawaiians with respect to developing a CD4 count < 200 cells/mm3. HSPAMM contains information on a large number of HIV-infected Asians/Pacific Islanders. PMID- 12632826 TI - Therapy with hypertonic saline in combination with anti-convulsants for hyponatremia-induced seizure: a case report and review of the literature. AB - Seizures are an uncommon but serious complication of hyponatremia which can lead to permanent brain damage and even death. It is recommended that patients with hyponatremic-induced seizures be treated with 3% hypertonic saline, however, a rapid rate of correction may result in central pontine myelinolysis (CPM), a severe neurological disorder characterized by mutism, dysarthria, spastic quadriparesis, and pseudobulbar palsy. The patient in this case developed a hyponatremia-induced generalized tonic-clonic seizure which was aborted by rapid therapy with diazepam, followed by hypertonic saline and phenytoin. Subsequent replacement of hypertonic saline with normal saline and salt tabs in combination with phenytoin allowed gradual correction of serum sodium without any subsequent seizures or neurological complications. PMID- 12632827 TI - Role of the library and librarians in medical education at the John A. Burns School of Medicine. PMID- 12632828 TI - Gynecologic cancers. PMID- 12632829 TI - [Image of the month. "The jewels of the crown"]. PMID- 12632830 TI - [How I manage patients developing thromboembolic complications as a result of breast cancer treatment with tamoxifen ]. AB - Thromboembolic complications are well known side effects of treatment with tamoxifen in patients with breast cancer. The authors review the pathophysiology and the risk factors that increase the probability to develop these complications. The most appropriate treatment is discussed. PMID- 12632831 TI - [Clinical case of the month. Coronary artery fistula]. AB - We report the case of a 54 year old man presenting atypical chest pain. The echocardiogram showed a dilatation of the coronary sinus. Subsequently the angiographic evaluation with cardiac catheterization demonstrated a coronary artery fistula between the circumflex artery and the coronary sinus and between a megaanevrysmal right coronary artery and the coronary sinus. No significant left to right shunt was discovered, however. PMID- 12632833 TI - [TVT: Revolutionary treatment of urinary incontinence]. AB - Stress urinary incontinence represents an important, often unknown and, yet, certainly most unpleasant pathology. Over years, several different surgical techniques have been proposed and reported to have variable success. TVT, a simple and reproducible technique, aims at stabilizing mid-urethra, and not bladder-neck. Our own clinical experience amounts to 139 cases. All these patients were evaluated by clinical examination and, subjectively, by a questionnaire. 89.2% were cured and 6.5% improved. The most frequent complication was bladder perforation (6.5%), but it had no incidence on the final results. Morbidity was low. This revolutionary technique is very promising and our own results are similar to those reported by others. PMID- 12632832 TI - [Diagnosis and treatment of pancreatic injuries in children]. AB - Pancreatic injury after trauma can occur in the child as an isolated lesion after a minor injury. The consequences can be severe in the absence of prompt diagnosis and treatment. Determination of mechanism of injury, clinical examination, blood amylase levels and abdominal CT-scan are first line steps for the evaluation of the injury. ERCP and MRI can be useful tools to further document injury to the pancreatic duct. In the absence of duct injury, conservative treatment can be applied. For the other cases, the treatment is surgical although newer techniques may be applied in selected cases. PMID- 12632834 TI - [Hepatic metastases of colorectal cancer: current therapies]. AB - Important progress has been made in the treatment of liver metastases of advanced colorectal cancer. Surgery with curative intent, when possible, shows evidence of prolonged survival. Response rate and overall survival can be improved with modern polychemotherapy. Cytotoxic drug combinations and sequential treatments sometimes make surgery possible for initially non resectable lesions. Impact of loco-regional treatment such as hepatic arterial infusion chemotherapy must be defined in randomised trials. Radiofrequency ablation is also currently evaluated in clinical trials. In this review the benefit of each treatment is discussed. PMID- 12632836 TI - [Asthenia and visual impairment in a 48-year old painter]. AB - Tiredness is a common complaint which may have multiple causes, sometimes difficult to ascertain. As occupational activities can play a major role in the incidence of various diseases, failure to investigate the patient's work history may hinder an adequate patient care. In the clinical case described hereafter, the patient symptoms could eventually be ascribed to a psycho-organic syndrome (POS) caused by the chronic exposure to organic solvents in the painting material used. This syndrome is non specific, evolves with time in an insidious manner, and, in its last stages, may cause non reversible brain damages. When facing non typical neurological symptoms in a working age adult, one has thus to consider the possibility of this diagnosis since the workers populations exposed to solvents are large in several occupations, and because the recent legal recognition in Belgium of this syndrome as an occupational disease does provide some interesting means for a better follow-up and protection of the patients concerned. PMID- 12632835 TI - [End of treatment evaluation in patients with aggressive non-Hodgkin's lymphoma]. AB - Incomplete regression of a lymphomatous mass after chemotherapy and/or radiotherapy constitutes a major problem in the treatment of lymphoma. In patients with persisting tumor, it could be reasonable to use salvage therapy and possibly hematopoietic stem cell transplantation at the time of minimal disease rather than at the time of clinically overt relapse. The authors reviewed the most appropriate imaging techniques for the assessment of response to treatment. The limitations of CT and MRI for predicting the nature of residual masses are well known. 67Ga scintigraphy has become a standard procedure for the posttreatment evaluation of patients with lymphoma, but it appears that 18F-FDG PET may be a more effective method. Personal experience in the field of PET scan is reported. Although PET should be considered the noninvasive imaging modality of choice, a histological confirmation of residual disease is always necessary before starting salvage therapy. 18F-fluorodeoxyglucose is not a tumor specific radiotracer. PMID- 12632837 TI - [Stromal tumor of the stomach with exo-gastric growth]. AB - A 60 year-old male patient presented with a giant stromal tumor of the stomach that showed predominant extragastric growth. Surgery consisted of total tumor resection with partial gastrectomy. The stromal origin of the tumor was established by immunohistochemistry. In spite of its large size and the presence of necrotic foci, the tumor was considered to be of low grade in view of the low mitosis index. The patient was maintained on careful surveillance. The histological features which characterize these tumors and their implications for prognosis and therapy are briefly discussed. PMID- 12632838 TI - [Prison psychosis and dissociative disorders]. AB - Through a few clinical case histories stemming from their daily activities at the psychiatric section of the Lantin Prison, the authors propose to revisit the classic concept of Prison psychosis. They broaden its limits to include other psychotic and dissociative phenomena common to the jail population. This requires a strict differential diagnosis, allowing to eliminate some similar pathologies; nevertheless, some difficulties and imperfections persist. The development of the psychosis, the input from the jail architecture and milieu, the predisposing as well as facilitating factors linked to the personality of the inmate, and triggering phenomena are discussed. Finally, the comorbidity between these psychotic/dissociative phenomena and the borderline & histrionic personality disorders is envisaged. PMID- 12632839 TI - [How I explore...gynecological pathology. Part 2: Supplementary examinations]. AB - The general decrease in the quality of anamnesis and clinical gynecological examination and the use of expensive complementary examinations lead us to redefine the bases of the gynecological outpatient consultation. The anamnesis and clinical examination have been detailed in the precedent issue of the Revue Medicale de Liege. The present article will explain the commonly performed complementary examinations during gynecological explorations. PMID- 12632840 TI - [The PROSPER Study (PROspective study of pravastatin in the elderly at risk)]. AB - Statins reduce coronary and cerebrovascular mortality and morbidity in middle aged individuals. Until recently, their efficacy and safety in elderly people had not yet been firmly established. PROSPER was a controlled, randomised study involving 2,804 men and 3,000 women aged 70-82, with a history of, or risk factors for cardiovascular disease. Their baseline cholesterol level was 135-350 mg/dl; they were randomised to either 40 mg pravastatin per day, or matching placebo. Average follow-up was 3.2 years. The primary endpoint was a composite of coronary death, non-fatal myocardial infarction, and fatal or non-fatal stroke. Pravastatin lowered LDL-cholesterol (-34%), and reduced the incidence of the primary endpoint (-15%; CI 95%: 3-26%; p = 0.014). Coronary death and non-fatal myocardial infarction risk was also reduced (-19%; p = 0.006), and mortality from coronary disease fell by 24% (p = 0.043). The risk for stroke, however, was unaffected (p = 0.8), whereas the incidence of transient ischemic attacks was reduced by 25%, which was (marginally) insignificant (p = 0.051). Pravastatin had no effect on cognitive functions or incapacity. New cancers were more frequent amongst pravastatin-treated individuals (+25%; p = 0.020). However incorporation of this new data in a meta-analysis of all pravastatin and all statin trials revealed no overall increase of cancer risk. PMID- 12632841 TI - [Pulmonary rehabilitation of COPD in Switzerland--an assessment of current status]. AB - The aim of this survey was to characterize respiratory rehabilitation programmes for patients with COPD in Switzerland. Rate of return of questionnaires was 94%. In 2001, 2238 patients with COPD followed an inpatient (1790) or outpatient (448) respiratory rehabilitation programme. This corresponds to 1% of all patients with COPD in Switzerland. Physical exercise is estimated to be significantly more important compared to breathing exercises, patient education and psychosocial support although this prioritization is not as pronounced as in the current literature. Disease-specific instruments to measure health related quality of life that represents the most important outcome measure for patients with COPD should be implemented in practice. Thereby both the components of rehabilitation programmes and predictive factors for a successful rehabilitation could be evaluated. PMID- 12632842 TI - [Bladder carcinoma--a current review]. AB - Bladder cancer is the sixth most common malignant tumor. The course of the disease depends largely on histological findings. A combination of transurethral resection and intravesical chemotherapy is indicated in superficial tumors and is able to cure most patients with a well differentiated cancer. Contrary to that, muscle-invasive and poorly differentiated tumors take a much less favorable course. They can be offered novel combined therapeutic approaches or/and at least orthotopic continent bladder substitution. It seems possible that the recently observed trend towards improved overall survival may continue. In the future, there should be more emphasis on prevention (elimination of carcinogens and early detection). PMID- 12632843 TI - [Ototoxicity of aminoglycoside antibiotics]. AB - In gram-negative infections aminoglycosides still remain to present a first-line antibiotic. Their use is limited by the high risk for side effects and especially nephrotoxicity and ototoxicity. The more recently developed members of this group, although purporting to be less ototoxic, are also capable of producing inner ear damage in sufficiently high doses. This paper reviews the morphological, histochemical, and electrophysiological evidence concerning the mode of action of aminoglycosides on the auditory and vestibular systems and discusses the available methods for the detection of possible clinical ototoxicity. With recent advances in aminoglycoside monitoring techniques, the risk of toxicity has been greatly reduced. Guidelines are proposed about the prevention of ototoxicity and the management of patients with established auditory or vestibular dysfunction. PMID- 12632844 TI - [Treatment refractory hypertension with atrophic kidney: attending to one and not neglecting the other]. AB - The incidence of primary aldosteronism in patients with difficult to treat hypertension is higher than previously stated in texts. The clinical picture of hypokalemic hypertensive patients remains unreliable as a third of the patients are normokalemic. Considering the diagnosis is the most important step in diagnosing primary aldosteronism. The clinical suspicion has to be confirmed biochemically. PMID- 12632846 TI - [From "inciting a public incident" to oncologic diagnosis. 49-year-old patient, former construction worker]. PMID- 12632845 TI - [Pancytopenia after viral infection in St. n. malignant lymphoma]. PMID- 12632847 TI - Peripheral arterial disease--Part II: Management. AB - While knowledge about peripheral arterial disease (PAD) appears well disseminated, the condition still goes largely undiagnosed and untreated. In PART 1 of this paper, we summarized the diagnosis, epidemiology and risk factors for PAD. Non-invasive treatment involves management of risk factors and targeted drug therapy. Invasive strategies include angioplasty, angiogenesis and surgery. Dealing with both the structural factors (physical plaque morphology) and functional ones (endothelial function) are important in effectively managing PAD. This paper (PART 2) deals with the management (medical and surgical) of PAD. PMID- 12632848 TI - Acupuncture: a physician's primer, Part II. AB - Acupuncture is one of the alternative therapies that is quickly gaining popularity and acceptance among both patients and doctors. The NIH has recently supported its use for many conditions, either alone or as adjunctive therapy to conventional methods. The World Health Organization (WHO) has recommended acupuncture worldwide for several decades and for numerous additional conditions. However, all acupuncture is not the same; training, licensure and experience vary widely, and conventional research support is lacking, all contributing to the success or failure of acupuncture treatment. With proper use, acupuncture can be an effective treatment for a variety of medical conditions, but used incorrectly, it can be a waste of time and resources. PMID- 12632849 TI - Leaders in medicine: Donald Mishler, MD. PMID- 12632850 TI - Influenza virus: natural disease and bioterrorism threat. PMID- 12632851 TI - The potential role of viral pathogens as agents of bioterrorism. PMID- 12632852 TI - Prevention of severe asthma. PMID- 12632853 TI - Living with asthma. PMID- 12632854 TI - Medicaid program information. PMID- 12632855 TI - Pitching plans to the uninsured. Insurers increasingly see viable market in growing pool of middle-class workers who don't have health insurance. AB - It's a bold new world in healthcare insurance. A few years ago, insurers would have been reluctant to use advertising dollars to reach the uninsured. But now, as the average income of the typical uninsured person is climbing and Congress is pondering initiatives to expand coverage, the market is gaining zealous suitors. "Things are really heating up," says Merrill Matthews Jr., left, director of the Council for Affordable Health Insurance. PMID- 12632856 TI - Supply and demand. AHA-FAH study: increased demand for services, high cost of labor factors behind rise in hospital spending. PMID- 12632857 TI - Doc-owned and -operated. PMID- 12632858 TI - Saving the day in Mississippi. PMID- 12632859 TI - A helping hand for his family. PMID- 12632860 TI - A measured dose of safety. Addressing adverse drug events is among first steps prescribed as Trinity health installs an electronic medical record system. PMID- 12632861 TI - Raising a red flag. Programs look for wrong combination of events. PMID- 12632863 TI - Second-line treatment of small-cell lung cancer. The case for systemic chemotherapy. AB - Small-cell lung cancer is an aggressive tumor associated with high rates of regional or distant metastases at diagnosis. Although highly chemosensitive to agents given in the first-line setting (e.g., etoposide and cisplatin), most patients relapse and have a poor prognosis. Treatment options for relapsed patients include radiotherapy for limited-stage disease and chemotherapy or combined modalities for advanced-stage disease. In clinical practice, however, some oncologists maintain that chemotherapy provides an insufficient survival benefit to justify the sometimes debilitating toxicity associated with the more active regimens in particular. Other potential barriers to further treatment include patient comorbidities, performance status, site(s) of progression, progression-free interval, and previous treatments. However, numerous clinical trials demonstrate that some patients benefit from treatment, achieving prolonged survival, symptom palliation, improved quality of life, and the opportunity, albeit rare, for durable remission. Additionally, several novel chemotherapeutics are available that alone or in combination help patients lead an improved quality of life. Finally, alternative routes and schedules--oral formulations, weekly administration, and prolonged treatment vacations--have been developed to deliver chemotherapy to patients with poor performance status or multiple comorbidities. This article reviews the advantages and disadvantages of treating recurrent small cell lung cancer and summarizes the utility of several active agents. PMID- 12632862 TI - Straight talk--new approaches in healthcare. Want better relationships with payors? Focus on medical management. AB - When it comes to managing medical care for their enrollees, payors are moving towards a broader focus for medical management that includes both costs and quality. In the twelfth installment of Straight Talk, we look at how health plans, such as CIGNA, are developing incentives to reward high-quality, low-cost providers of medical care and transitioning their utilization management operations to a new model. To ensure quality, they are beginning to compare providers' performance to national standards for medical outcomes. Through these initiatives, known as "medical management," payors hope to improve not only medical care but also their relationships with providers. PMID- 12632864 TI - Testicular cancer: maintaining the high cure rate. AB - The management of germ cell tumors has advanced dramatically, with cure rates approaching 90% to 95%. Treatment of stage I/A seminomas generally includes orchiectomy and adjuvant radiotherapy. Treatment of stage I/A nonseminomatous germ cell tumors involves orchiectomy followed by retroperitoneal lymph node dissection or active surveillance. One of the major advances has been the introduction of cisplatin-based chemotherapy for metastatic disease and the development of a system of risk attribution. The logical management of any patient with curable disease is to provide curative therapy and then follow the patient in a structured manner, to diagnose and treat any complications in a timely manner. PMID- 12632865 TI - Clinical trials referral resource. Current clinical trails in endometrial cancer. PMID- 12632866 TI - Current status of monoclonal antibody therapy for chronic lymphocytic leukemia. AB - Despite many therapeutic options for chronic lymphocytic leukemia (CLL), the disease remains incurable. Since monoclonal antibodies and recombinant toxins that bind surface antigens expressed on the malignant lymphocytes have been developed, targeted therapy has become a vital option in treating CLL. Rituximab (Rituxan), a chimeric human-mouse anti-CD20 antibody, and alemtuzumab (Campath), a humanized anti-CD52 monoclonal antibody, have both shown activity in CLL--as single agents and in combination with conventional chemotherapy. The possibility of combining antibodies has been explored as well, with some efficacy. In this review, we discuss the clinical data on the activity of commercially available antibodies in CLL, both as monotherapy and in combination with other agents. PMID- 12632868 TI - Why should we care about the cancer anorexia/weight loss syndrome? PMID- 12632867 TI - Myalgias and arthralgias associated with paclitaxel. AB - Paclitaxel-induced myalgias and arthralgias occur in a significant fraction of patients receiving therapy with this taxane, potentially impairing physical function and quality of life. Paclitaxel-induced myalgias and arthralgias are related to individual doses; associations with the cumulative dose and infusion duration are less clear. Identification of risk factors for myalgias and arthralgias could distinguish a group of patients at greater risk, leading to minimization of myalgias and arthralgias through the use of preventive therapies. Optimal pharmacologic treatment and possibilities for the prevention of myalgias and arthralgias associated with paclitaxel are unclear, partially due to the small number of patients treated with any one medication. The effectiveness of nonsteroidal anti-inflammatory drugs (NSAIDs) is the most frequently documented pharmacologic intervention, although no clear choice exists for patients who fail to respond to NSAIDs. However, the increasing use of weekly paclitaxel could necessitate daily administration of NSAIDs for myalgias and arthralgias and leave patients at risk for adverse effects. This concern may also limit the use of corticosteroids for the prevention and treatment of paclitaxel-induced myalgias and arthralgias. Data from case reports suggest that gabapentin (Neurontin), glutamine, and, potentially, antihistamines (e.g., fexofenadine [Allegra]) could be used to treat and/or prevent myalgias and arthralgias. Given the safety profile of these medications, considerable enthusiasm exists for evaluating their effectiveness in the prevention and treatment of paclitaxel myalgias and arthralgias, particularly in the setting of weekly paclitaxel administration. PMID- 12632869 TI - Supportive care in cancer: a general overview. PMID- 12632870 TI - Nutrition as an integral component of supportive care. PMID- 12632871 TI - Specialty nutrition: new approaches and novel ingredients. PMID- 12632872 TI - Patient-generated subjective global assessment. PMID- 12632874 TI - Special considerations for nutrition intervention with oncology patients. PMID- 12632873 TI - PanCAN PALS--patient and liaison services: an innovative model for informed patient decision-making, including patient information and clinical trials. PMID- 12632875 TI - Excellence in patient management. PMID- 12632876 TI - The precious pill: access by the aged to affordable pharmaceuticals. PMID- 12632877 TI - Using high-intensity care management to integrate acute and long-term care services: substitute for large scale system reform? AB - This study evaluates a demonstration that used high intensity care management to improve integration between the acute and long-term care service systems. The demonstration intervention included the use of clinical nurse care manager, supervised by a geriatrician, to supplement an existing in-home care management system. Chronically disabled home care clients age 60 and over were randomly assigned (N = 308) to receive enhanced clinical services plus traditional care management, or to the control group, to receive the normal care management services provided. Treatment group members were expected to experience lower use of hospitals and nursing homes and lower overall health and long-term care costs. Research subjects were followed for up to 18 months using Medicare records and mortality data. A subsample (N = 150) also received in-person interviews to cover a range of health and social outcomes anticipated as a result of the intervention. Although there was some variation in health use and cost across treatment and control groups over the 18 month time period, the overall conclusion is that there were no differences between groups on any of the outcome variables examined. Efforts to integrate the acute and long-term care systems have proven to be difficult. This intervention, which attempted to create integration through high intensity care managers, but without financial or regulatory incentives, was simply unable to create enough change in the care system to produce significant change for the clients served. PMID- 12632878 TI - The case manager's view of the helping alliance. AB - Although the helping alliance has been a topic of investigation in psychotherapy research for decades, few studies have examined the role of the helping alliance in assertive community treatment programs serving people with severe mental illness. In this article, we describe a series of analyses focused on the case manager's view of the helping alliance. The study addressed two primary questions: What factors facilitate a positive helping alliance in case management? What is the relationship of the helping alliance to client outcomes? Results indicated that few client variables predicted the helping alliance. Providing a wider range of services was associated with a stronger alliance at months but not at 18 months. A strong alliance at 18 months was associated with a strong alliance at 6 months. In addition, the alliance at 18 months was rated more positively for clients who were African-American and who did not have a substance use disorder. The helping alliance predicted client satisfaction, anxiety-depression, and hostility-suspicion outcomes at 6 months. However, the helping alliance did not predict any of the client outcome variables at 18 months. PMID- 12632879 TI - Putting the costs of prescription drugs in perspective for Medicare beneficiaries. PMID- 12632880 TI - Costs and benefits of pharmaceuticals: the value equation for older Americans. AB - Recent increases in drug expenditures are primarily due to the availability of more and better therapy rather than price inflation. Investment in new drugs generates savings throughout the health care system. Increased use of drugs, especially newer agents, has also resulted in increased longevity and reduced disability. Benefits from new pharmaceuticals for outweigh their costs for many key diseases of the elderly. Even incremental improvements in drug therapies contribute substantially to improved care. Chronic illness, disability, and an aging population will drive future health care spending. Pharmaceutical innovation will be an integral part of effective strategies to address this challenge. The availability of individualized therapy for the elderly will soon increase based on our rapidly growing understanding of the molecular and genetic basis of disease. This is expected to result in major advances in preventing, treating, and perhaps even curing many of the costly, life-threatening, and disabling diseases afflicting older Americans. The adequacy of drug benefit programs for elderly patients depends on the extent to which the range of drug therapies necessary for appropriate care are covered. Policies that foster the availability of unique pharmaceuticals can have important implications for treatment outcomes, quality of life, cost containment, and ongoing research investment in newer and more effective medicines. Such policies increase the diversity of agents within drug classes and thereby enable differentiated, individualized therapy. A wide range of choices is especially important for elderly patients, who have the greatest need for individualized care and are at greatest risk for compromised outcomes if choices are overly circumscribed. PMID- 12632881 TI - The high price of prescription drugs and its impact on the elderly. PMID- 12632882 TI - New York State's EPIC program: helping seniors afford the rising cost of prescription drugs. PMID- 12632883 TI - [Ischemic heart disease and gastro-oesophageal reflux disease--simultaneous 24 monitoring of ECG and oesophageal pH]. AB - AIM: To determine the correlation between episodes of ischaemia in ECG recordings with pathologic gastro-esophageal reflux during simultaneous 24 hour monitoring of ECG and oesophageal pH. METHODS: Simultaneous 24 hour monitoring of ECG and oesophageal pH was performed in 30 patients (p) (26 M/4F, aged 39-74) with coronary artery disease of CCS class II-III, CAD was confirmed in coronary angiography. Analysis of the oesophageal pH was performed by using the Polygram programme (PW-version 2.04 Esophogram-version 2.01). ST depression > 1 mm and lasting at least 1 min was regarded as significant in ECG monitoring. Pathologic gastro-esophageal reflux was defined as a drop in pH < 4 lasting more than 5 min. Gastrooesophageal reflux disease (GERD) was diagnosed when a drop in pH < 4 lasted for more than 5% of the monitoring period. Gastro-oesophageal reflux dependent ST depression was defined as an ST depression that occurred during reflux episode and lasted up until 10 min from the end of the reflux. RESULTS: 26 patients (87%) had a total of 116 episodes of ST depression and 21 out of the 116 episodes (18%) were Gastro-oesophageal reflux time dependent. Fifteen patients (50%), had at least one episode of ST depression, depending on the time of reflux. Pathologic gastro-esophageal reflux was present in 25 patients (85%). In 14 patients (46.6%), the GERD pH criteria were fulfilled. In this group of patients, there was a significantly longer time of total ST depression (total ischaemic burden). CONCLUSIONS: 1. GERD is a frequent disease in patients with angiographically proven coronary artery disease. 2. Pathological gastroesophageal reflux can induce myocardial ischaemia, which can be determined by analysis of ST depression during 24 hour monitoring of ECG. PMID- 12632884 TI - [Dobutamine stress echocardiography in symptomatic patients with non-diagnostic or negative treadmill test]. AB - In patients suffering from angina like exertional pain or dyspnea with a negative or undiagnosed exercise treadmill test (u/n TET), controversy exists regarding indications to perform coronary angiography. This study was performed in order to evaluate the diagnostic significance of dobutamine echo-stress (DSE) in assessing the possible causes of symptoms in such patients. We retrospectively analysed 550 patients. Out of this number, 128 patients had a u/nTET (study group--SG) and 422 patients had a positive TET--control group (CG). The exercise treadmill test was performed according to the Bruce protocol. Dobutamine stress echocardiography was performed according to a standard protocol. RESULTS: In patients that were DSE(+), no significant differences in the analysed parameters were found between study and control groups. In patients that were DSE(-), monovariate analysis detected that gender, age, history of hypertension, LVH, sigmoid IVS, small LV and EF as factors predicting clinical symptoms. Multivariate analysis revealed that middle-aged women, elderly men, and history of hypertension were independent predictors of exertional symptoms. Multivariate analysis also showed that intracavitary LV obstruction or LV outflow tract obstruction in hypertensive females was an independent predictor of reproducibility of symptoms. CONCLUSIONS: 1. Angina-like symptoms in patients with non-diagnostic/negative TET and negative DSE depends on several clinical and echocardiographic factors. 2. Dynamic intracavitary LV obstruction or LV outflow tract obstruction, especially with a hypotensive reaction, are potential causes of symptoms in patients with non diagnostic/negative TET and negative DSE. 3. Reproducibility of symptoms during negative DSE occurred in hypertensive females with dobutamine-induced intracavitary LV obstruction or LV outflow tract obstruction. PMID- 12632885 TI - [Role of the right chest lead in improving the accuracy of myocardial ischemia detection]. AB - BACKGROUND: The limitations of exercise electrocardiography in the detection of coronary artery disease includes low sensitivity, especially in isolated right coronary artery (RCA) disease. Recent studies postulated the addition of right precordial leads to the standard exercise ECG (ExT) recording for better detection of right ventricular ischemia. PURPOSE: We tested the hypothesis that the replacement of the standard first precordial lead (V1) with a fourth right chest lead (V4R) can improve the diagnostic accuracy of ExT. MATERIALS AND METHODS: We studied 100 patients (78 men and 22 women, aged 55 +/- 9 years (31 71)) who underwent treadmill ExT (Bruce protocol) and coronary angiography. 52 patients had the V4R lead (Group V4R) instead of V1. The control group (Group V1) included 48 patients with the standard 12-lead ECG. Hemodynamically significant lesions were defined as stenosis > = 50% in left main coronary artery or > = 70% in other arteries. RESULTS: 76 out of 100 patients had significant coronary lesions on the arteriogram, 81% in Group V4R and 71% in Group V1. Overall sensitivity (71% Group V4R vs 79% Group V1), specificity (40% Group V4R vs 21% Group V1) and accuracy (65% Group V4R vs 63% Group V1) of ExT for detection of significant coronary artery disease was similar in both groups. There was also no statistically significant difference in the ability to define an inferior or right ventricle vs. anterolateral ischemia between both groups. Only 5 out of 32 (16%) patients with significant lesions in RCA revealed ST changes in V4R. CONCLUSIONS: In our study, the application of V4R instead of V1 did not significantly change the accuracy of ExT. Typical ECG signs of ischemia in V4R have high specificity, but low sensitivity for RCA disease. Thus, modification of the standard 12-lead scheme cannot be advocated for routine ExT, however although the use of a wider electrocardiographic window (e.g. V3R-V5R) might be more successful for recording right ventricular ischemia. PMID- 12632886 TI - [The influence of percutaneous coronary recanalization of total coronary occlusions on the heart rate variability ]. AB - In patients following a myocardial infarction, heart rate variability is an important prognostic factor. Decreased heart rate variability is associated with a higher mortality rate. This study evaluated the influence of recanalisation by percutaneous coronary intervention (PCI) in totally occluded coronary arteries on the heart rate variability in patients with myocardial akinesis in the recanalised artery area. The study group included 22 men (average age 52.8 +/- 7.7) after successful PCI recanalization of the totally occluded artery. All patients had akinesis in the recanalised artery area. All patients underwent 24 hour continuous electrocardiographic Holter monitoring with HRV analysis and echocardiographic dobutamine tests both before the PCI and 6 months afterwards. The population was divided into two groups: group A (10 patients) had contractility adjustments in echocardiographic tests that were performed 6 months after the PCI. Group B (12 patients) did not have contractility adjustments. Before the PCI, there were no differences in the wall motion score index (WMI) or the time-domain HRV parameters between the two groups. After 6 months, there was a significant increase in the WMI value in group A, whereas this index remained unchanged in group B. In group A the time-domain HRV parameters had improved 6 months after the PCI, whereas they remained unchanged in group B. Successful percutaneous revascularization of the totally occluded coronary artery in patients with myocardial akinesis in the occluded artery area leads to an increase in HRV parameters. This improvement occurred only in patients that had a contractility adjustment after the PCI in previous akinetic segments of myocardium. PMID- 12632887 TI - Nitric oxide stimulation of cardiac pacemaking in the sino-atrial node through the activation of a novel signalling pathway: overview of in vitro and in vivo evidence for a new basic mechanism in the control of heart rate. AB - Recent work has tested the hypothesis that nitric oxide (NO) might play an important role in the control of heart rate (HR) by stimulating cardiac pacemaking in the sino-atrial node (SAN). In isolated guinea-pig SAN cells and atrial preparations, nano-micromolar concentrations of NO donors were shown to elicit a NO-dependent increase of 20-30% in the beating rate. In the pig heart in situ, topical NO donor application to the SAN increased HR by ca. 10%, indicating that the direct positive chronotropic effect of NO is relevant in vivo. Indeed, a similar significant increase in HR (by 12%) was seen in humans with intravenous NO donor infusion while blood pressure was clamped. Inhibition of NO synthase or depletion of its gene have been associated with a significant reduction in HR by ca. 10% in vitro and in vivo, consistent with a tonic positive chronotropic effect of endogenous NO. The direct stimulatory effect of NO on cardiac pacemaking is mediated by stimulation of the 'pacemaker' current I, in SAN cells via a novel cascade involving cGMP, PDE3-PKA signalling and mobilization of intracellular calcium. In conclusion, recent studies have demonstrated for the first time that NO exerts a direct positive chronotropic effect through the activation of a novel signal transduction pathway in SAN cells. Evidence in animals and in humans indicates that direct stimulation of cardiac pacemaking by NO is functionally relevant for the control of HR in vivo. PMID- 12632888 TI - [Heart rate variability in patients treated with percutaneous transluminal coronary angioplasty]. AB - Decreased heart rate variability (HRV) is an independent risk factor for sudden cardiac death in patients with coronary artery disease (CAD) and previous myocardial infarction. The influence of myocardial revascularization on HRV parameters has been investigated for the past few years. However, little is known about HRV behaviour in patients with CAD treated with percutaneous transluminal coronary angioplasty (PTCA). The purpose of this study was to estimate HRV parameters before PTCA and during a one year follow-up. We examined 65 patients (47 men, 18 women; mean age 51.2 +/- 7.6), who underwent successful and uncomplicated PTCA. Thirty eight patients had a prior myocardial infarction. The mean left ventricular ejection fraction was 59.8 +/- 8.1%. Thirty-nine patients had a single-vessel disease, 25 patients had a double-vessel disease and one patient had a triple-vessel disease. Serial 24-hour Holter monitoring (before PTCA, 14 days, 3, 6, 12 months after PTCA) was performed in all patients. The following time domain measures of HRV were calculated from each recording, for 24 hours: pNN50, SDNN, SDANN and rMSSD. During the one year follow-up, a significant increase of pNN50, SDNN and SDANN was observed. We noted, that the statistically significant increase of the above-mentioned parameters occurred during the first 3 month follow-up. The results of the next serial recordings (6 and 12 months after PTCA) demonstrated no additional changes in HRV values, however they were still significantly higher than before procedure. Mean values of HRV parameters before PTCA and 3 months after were as follows: pNN50: 8.8 +/- 6.8%, vs 11.7 +/- 9.4% (p < 0.05), SDNN: 121.2 +/- 33.5 ms vs 148.2 +/- 29.3 ms (p < 0.05), SDANN: 113.6 +/- 16.2 ms vs 125.1 +/- 22.3 ms (p < 0.05). Parameter rMSSD did not change significantly during the whole observation. CONCLUSIONS: 1. Myocardial revascularisation with successful PTCA leads to an improvement in the autonomic modulation of heart rate. 2. The increase in parasympathetic control of the heart rate is already being observed 2 weeks after successful PTCA. 3. Beneficial increased influence of both parts of the autonomic nervous system on heart rate persists for 12 months after coronary angioplasty. PMID- 12632889 TI - [Heart rate variability in patients after coronary artery bypass grafting--early and long term effects of cardiac rehabilitation]. AB - Patients after coronary artery bypass grafting (CABG) have impaired heart rate variability (HRV) parameters. The clinical significance of impaired HRV and the influence of the cardiac rehabilitation on the HRV in this group of patients is not clear. We studied 103 patients (average age 52.2 +/- 7.8) after CABG, divided into group A (72 patients) who underwent cardiac rehabilitation (starting 15 days after the CABG) and group B (31 patients) who did not have rehabilitation. In all patients, a 24 hr ECG (including time and frequency domain HRV) was obtained before the onset of rehabilitation, after completion of the rehabilitation and after 6 months. Before the rehabilitation there were no differences in time domain HRV between the groups, however frequency-domain HRV values were significantly higher in group A. In group A, the time-domain HRV parameters improved after 21 days of rehabilitation, whereas the frequency domain parameter components remained unchanged. At 6 months, improvement in time and frequency domain HRV parameters was noted, but the parameters were significantly higher in group A. In patients that underwent coronary artery bypass grafting, the cardiac rehabilitation time domain HRV showed improvement in both the short term (21 days) and long run (6 months). However, the frequency domain parameter was only increased over the long run. Cardiac rehabilitation speeds up the improvement of the time domain HRV and frequency domain parameters over long term observation (6 months). PMID- 12632890 TI - [Heart rate variability in patients with hypertension and left ventricular hypertrophy]. AB - The aim of the study was to analyse the heart rate variability (HRV) in patients with moderate essential hypertension complicated by left ventricular hypertrophy (LVH). Forty-two patients with untreated essential hypertension participated in the study group and 45 normotensives (16M, 29F, age 49.8 +/- 5.6 years) served as the controls. The hypertensives were divided into those with LVH (n = 25, 10M, 15F, age 50.1 +/- 5.9 years) and those without LVH (n = 17, 4M, 13F, age 52.0 +/- 3.4 years). HRV was recorded and analysed using the CardioPSA System, Medatec. In each subject, 30 min. of ECG recording was obtained: 15 min. in the supine, position and 15 min. in the upright position. Fast Fourier transform was used to analyse total power, low frequency (LF, 0.04-0.15 Hz) and high frequency components (HF, 0.15-0.40 Hz). Left ventricular mass was calculated from echocardiograms using the Deveraux formula. LVH was defined as a left ventricular mass index (LVM) > 134 g/m2 for men and > 110 g/m2 for women. A T-test was used to test for nter-group differences: p < 0.05 was considered as significant. In hypertensive patients with left ventricular hypertrophy, we observed decreased values of HF component, both in supine and upright positions, as compared to hypertensives without LVH. Components of HRV did not differ between the hypertensives without LVH and the normotensives. In hypertensives with a LVH the high frequency component of HRV was decreased, indicating impaired para sympathetic activity. PMID- 12632891 TI - [The importance of echocardiographic evaluation in patients treated with cardiac pacing]. AB - This study evaluated the usefulness of echocardiographic evaluation in the selection of optimal atrioventricular delay (AVD) in DDD-paced patients. We discussed the influence of various AVD programming on systolic and diastolic left ventricle function. The detrimental effect of diastolic mitral regurgitation (DMR) on stroke volume was emphasized. Clinically useful echocardiographic methods of optimal AVD selection and prevention of DMR was discussed. PMID- 12632892 TI - Characteristics of patients with sinus node dysfunction. PMID- 12632893 TI - [Atrial refraction in patients with paroxysmal atrial fibrillation]. PMID- 12632894 TI - [Clinical implications of different locations of atrial septal defects]. AB - A frequent congenital heart disease in adults is the atrial septal defects (ASD). The pathology represents substantial variability due to the different mechanisms of developmental failure that may occur during embryogenesis. Subsequently, the clinical manifestation of the defect varies despite some similarities. This article presents classification, prevalence, clinical manifestation of various forms of atrial septal defects and scope of available treatment. PMID- 12632895 TI - [Echocardiographic assessment of early and midterm results of transcatheter closure of a narrow patent ductus arteriosus in children]. AB - This paper presents the echocardiographic assessment of early and midterm results of transcatheter closure of a narrow patent ductus arteriosus (PDA) in children using Jackson and Gianturco coils. The preliminary criterion for qualifying patients for the procedure was the PDA diameter (< or = 4 mm) at the narrowest site, as assessed echocardiographically. The criterion was met by 22 of 116 children with PDA treated at the Department of Pediatric Cardiology, Faculty of Medicine, Jagiellonian University, between January 1, 1996 and December 31, 2000. Angiography revealed, that in one case, the PDA diameter was larger (6.5 mm) and the patient required two Gianturco coils. Of 22 patients (8 males and 14 females) aged 2.7-16 years (x = 6.5 +/- 3.8), the procedure was successful in 21 children (95.5%). One patient with PDA was referred to the surgical department due to complications (the coil was displaced to the periphery of the left pulmonary artery and required removal by an Amplatzer device). The analysis of the follow up echocardiography performed within 48 hours after the procedure and 1, 6, 12, 24 and 48 months later, revealed immediate total PDA closure in 29% of children. In 33% of patients, the PDA closed within 24 months. One child did not report to follow-up, however, the others are asymptomatic and show residual negligible shunts. The authors determined that the incidence and time of closure of residual shunts is dependent on the magnitude of the left-to-right shunt before the procedure. PMID- 12632896 TI - [Semiautomatic algorithm for tracing an outline of the endocardium in newborns]. AB - Visual assessment of the endocardial outline is too subjective and the results do not provide enough support for reliable analysis. The main problem that hinders echocardiographic diagnosis in measurements of ventricular size and shape during subsequent cycles of cardiac function lies in difficulties in obtaining precise delineation of endocardial margins. A new, semiautomatic algorithm for tracing endocardial outlines of newborns that were detectable in echocardiographic images is presented. The material consisted of 1920 historical echograms of newborns with selected congenital heart defects (total anomalous pulmonary venous return- TAPVR, hypoplastic left heart syndrome--HLHS, and transposition of the great arteries--TGA) and control group. The algorithm was successfully used in the analysis of 384 images of heart chambers (right and left ventricles) of newborns in systole and diastole. The results were subjected to a statistical analysis and revealed that semiautomatic algorithms can give consistent, repeatable, good quality results, and image analysis methods can be an efficient, additional tool for the quantitative interpretation of echocardiograms of the endocardium in newborns. PMID- 12632897 TI - [The influence of increased preload on left ventricular systolic function in infants with congenital heart disease]. AB - The aim of the study was to assess the systolic function of the volume overloaded left ventricle (LV) in infants with congenital heart disease. MATERIAL: Seventy five consecutive patients (36 males, 39 females) with following breakdown were examined: 14 (18.7%) newborns, 24 (32%) infants under 3 months, 16 (21.3%) between 4 and 6, and 21 (28%) infants above 6 months of age. VSD was recognized in 60 (80%), PDA--6(8%), DORV--7 (9.3%), and CT (type I) in 2 (2.7%) of the patients. METHODS: Based on ECHO studies LV end diastolic volume (LVedv), end diastolic muscle mass (LVedm), LV shortening fraction (FS), ejection fraction (EF), LV fractional area changes (FAC), and heart rate (HR) versus pulmonary blood flow (Qp), which reflected the LV preload systemic blood flow (Qs), Qp/Qs, Qp-Qs (left-to-right shunt) and indexed for body surface area (BSA) and age, were assessed by statistical analysis. RESULTS: With a gradual increase of BSA with age (cc = 0.82, p < 0.001), a significant decrease in Qp-Qs (cc = -0.27, p = 0.035) and Qp/Qs (cc = -0.31, p = 0.013) were noted. Qp for the whole group ranged from 3.3 to 21.7 l/min/m2 (m = 8.9, x = 9.7, SD = 4.1), significantly increased with the rise of Qp-Qs (cc = 0.9, p < 0.0001) and influenced the increment of Qp/Qs (cc = 0.59, p < 0.001). Qs for the whole group ranged from 2.1 to 11.7 l/min/m2 (m = 4.3, x = 4.6, SD = 1.7) correlating with the rise of Qs (cc = 0.45, p < 0.001), and being lower than 2.5 l/min/m2 in only 5 (6.7%) patients but in only 5 (6.7%) patients (all newborns). The heart rate gradually decreased with age (r = -0.26, p = 0.04), from mean 145/min in newborns to 123/min in infants over 6 months, and with BSA (cc = -0.34, p = 0.006). Heart rate also correlated well with Qp (cc = 0.31, p = 0.014). The increase of LVedv was related to age (cc = 0.37, p = 0.003), BSA (cc = 0.43, p = 0.0004) and Qp (cc = 0.33, p = 0.009). However, only in newborns was the mean LVedv within normal limits for age. LVedm only correlated with BSA (cc = 0.26, p = 0.04), except in newborns where its mean values exceeded the upper limits for age in other subgroups of patients. LV systolic function indicators (FS, EF, FAC) were diminished in only 2 cases and did not correlate either with age, BSA, nor with hemodynamic parameters. CONCLUSIONS: 1. The reactions to increased preload of the systemic ventricle in children below one year of age were different. In newborns, the adrenergic response predominated, while infants manifested increased end diastolic volume and left ventricular muscle mass. 2. The systolic function of the overloaded left ventricle in infants was normal regardless of age, body surface area and intensity of hemodynamic disturbances. PMID- 12632898 TI - [The role of transesophageal echocardiography (TEE) in assessment of left ventricular tract obstruction]. AB - Twelve children with subaortic stenosis were analysed. Nine of them developed left ventricular outflow obstruction after surgery for congenital heart disease (VSD + IAA, VSD + DORV, VSD + TGA, VSD + CoA) and then developed an isolated form of primary stenosis. Both echocardiographic exams, TTE and TEE were performed in all patients. TTE was sufficient to assess isolated subaortic stenosis. In children after cardiosurgery, TEE was more reliable and provided more detailed visualisation of the stenosis and its relationship to surrounding structures. PMID- 12632899 TI - Preliminary cardiological examinations in deaf children. AB - There is some evidence that deaf children are more threatened than the general population by dangerous heart arrhythmias. An example is Jervell-Lange-Nielsen syndrome (one of the forms of long QT syndrome) which is characterised primarily by congenital deafness and prolongation of the QT interval. The aim of this study was to perform preliminary cardiological examinations on 162 deaf children (76 girls and 86 boys, 3-15 years old, mean age 10.5 +/- 2.8 years) who attend the Regional School Centre for Deaf Children in Katowice. The data in our analysis was obtained from case histories (school records and special questionnaires sent to parents), physical examinations with special regard to the cardiovascular system, double blood pressure measurement and 12 lead surface electrocardiogram. In the studied group, 90 children (55.5%) were congenitally deaf. Within this group, 24 children manifested tachycardia while 4 children manifested bradycardia. A-V block of I degree was found in 3 children. Incomplete right bundle branch block (RBBB) was observed in 44 children while a complete RBBB was noticed in 1 child. QTc interval was prolonged (> 0.44s) in 12 children suffering from congenital form of deafness and in 16 children suffering from all causes of deafness. On the basis of case history, physical examination, and standard 12 lead electrocardiogram, we established the occurrence of risk factors according to the new diagnostic criteria of Schwartz et al. with relative points assigned to the electrocardiographical, clinical and familial findings. These points when summed up represent the risk of sudden death caused by ventricular arrhythmia due to long QT syndrome. In the studied group (according to the Schwartz criteria), 2 (1.2%) children had high probability (> or = 4 points) of long QT syndrome (LQTS), 25 (15.4%) children had intermediate probability of LQTS (2-3 points) and 135 children had low probability of LQTS (< or = 1 point). PMID- 12632900 TI - [Diagnostic problems in recognizing structural pathology of the heart during the prenatal period]. AB - The aim of our study was to analyse the results of echocardiographic examinations of 28 fetuses with structural heart abnormalities referred for echocardiographic examination from January 1996 to May 2001 along with their follow-up. Fetal heart anomalies were diagnosed in 2.1% of all of the examined fetuses (28/1319). Among 28 cases of structural anomalies, there were 23 cases of heart defects, 4 cases of cardiomyopathy, and one case of intracardiac tumors. In 5 cases, heart defects were accompanied by arrhythmias while in 2 cases, heart defects were accompanied by congestive heart failure. Extracardiac malformations coexisted with heart defects in 11 fetuses. Eight newborns were operated shortly after birth. The mortality rate in the examined group was 64%. PMID- 12632901 TI - [Evaluation of surgical treatment results of coarctation of the aorta in neonates and young infants]. AB - The authors present the analysis of surgical results obtained for coarctation of the aorta (CoA) in children less than 1 year of age. The material consisted of 103 infants (62 Males and 41 Females) aged 0.33-10.5 months (x = 3.3 +/- 2.6 months) treated between January 1, 1985 and December 31, 1999. All the patients were subjected to a detailed physical examination, ECG, chest X-ray and comprehensive echocardiography, while 12 children had additional hemodynamic studies and angiocardiography. Two groups were distinguished among the investigated children: Below 3 months of life (Group 1, N = 65) and above 3 months of life (Group 2, N = 38). In 69 infants, (including 55 from Group 1 and 14 from Group 2), the recommendation for surgery was circulatory failure, while in 34 infants (10 from Group 1 and 24 from Group 2), the recommendation for surgery considerable systemic hypertension with absent femoral pulses. In 76 patients, the Waldhausen procedure was performed, while 24 were subjected to aortic isthmus angioplasty using a Gore-Tex patch and 3 were subjected to end-to end anastomosis. Six patients died, including 5 from Group 1 and 1 from Group 2. The remaining 97 infants (60 from Group 1 and 37 from Group 2) were followed-up for a mean period of 96.2 +/- 48.2 months. Postoperative recoarctation was encountered in 12 patients (12.4%); the condition was more predominant in Group 1 (16.6%) than in Group 2 (5.4%), but, no statistical significance was noted (Chi 2 = 2.677, p = 0.102). Despite the repair of aortic coarctation, systemic hypertension was noted in 17 children (17.5%) and it was equally common in both groups, but, twice as frequent in children with recoarctation. CONCLUSIONS: 1. CoAo correction in children below 3 months of age is associated with an increased risk of recoarctation. 2. Even when surgical treatment of CoAo is attempted very early in life, the risk of elevated systemic blood pressure is not completely eliminated. PMID- 12632902 TI - [Cardiopulmonary exercise capacity in adult patients with atrial septal defect]. AB - The aim of the study was to evaluate cardiopulmonary exercise capacity (WTS) in adult patients with atrial septal defect (ASD). After excluding 10 patients with additional cardiac, pulmonary or muscle diseases, 53 patients with ASD (Gr-ASD; mean age 37.8 +/- 14.9 years; 35 women, 19 men) were enrolled in our study. In addition, we evaluated 22 healthy age and gender matched control subjects (Gr-K; mean age 36.6 +/- 14.9 years; 13 women, 9 men). First, all subjects underwent resting spirometry and forced vital capacity--(FVC; % of predicted value), one second forced expiratory volume--(FEV1; % of predicted value) and FEV1/FVC (%) were determined. Then each subject performed a symptom-limited, incremental exercise test (modo Bruce). We evaluated the following parameters of resting metabolism: respiratory rate--(RR; L/min), minute ventilation--(VE; L/min), tidal volume--(Vt; L), oxygen uptake--(VO2; ml/kg/min), end-tidal carbon dioxide pressure--(PET CO2; mmHg), end tidal oxygen pressure--(PET O2; mmHg), ventilatory equivalent for carbon dioxide--(VE/VCO2) and ventilatory equivalent for oxygen- (VE/VO2). The following exercise parameters were analysed: peak oxygen uptake- (VO2 peak; ml/kg/min), VO2 peak expressed as % of predicted value--(VO2 %N), anaerobic threshold--(AT; % VO2 max), ventilatory equivalent for carbon dioxide- (VE/VCO2), end-tidal carbon dioxide pressure--(PET CO2; mmHg), O2 pulse, time of exercise--(T; min) and time to AT--(TAT; min). In addition, we performed a subgroup analysis for ASD patients below and > or = 40 years of age. All values were expressed as mean +/- SD. RESULTS: Although there was no significant difference in FVC, FEV1, FEV1/FVC, VO2, VE, Vt, RR between ASD patients and controls, the ASD patients aged 40 or older showed significantly lower FEV1/FVC and significantly higher RR. Expiratory gas exchange analysis showed significant differences between the ASD patients and controls (higher VE/VCO2, VE/VO2, PET CO2, and lower PET O2 respectively). Moreover, VO2 peak, VO2%N, AT, PET CO2 and O2 pulse were significantly lower while VE/VCO2 was significantly higher in ASD patients than in normal subjects. On average, VO2%N in ASD patients was only 61% of the predicted value for age and sex. Total time of exercise and time to AT were significantly shorter in ASD patients as well. CONCLUSIONS: WTS allows appropriate evaluation of cardiopulmonary capacity in ASD patients, though the mechanisms underlying ventilatory and hemodynamic abnormalities are still not fully understood. Adult patients with ASD reveal ventilatory abnormalities with an age-related trend towards deterioration. Cardiopulmonary exercise capacity in adults with ASD is markedly reduced in comparison with healthy population and deteriorates with age. WTS complements echocardiographic and hemodynamic evaluation of patients with ASD, and is helpful in indicating patients for ASD closure and their follow up. PMID- 12632903 TI - Circadian heart rate variability in patients with primary arterial hypertension. AB - INTRODUCTION: Abnormalities of autonomic nervous system function (ANS) exist in patients with hypertension and have been considered as one of the important factors in developing of essential hypertension. However the information about the effect of antihypertensive treatment (angiotensin-converting enzyme inhibitors--ACEI) on cardiac ANS activity is scarce. The main aim of our study was to evaluate circadian heart rate variability changes in primary hypertension treated with ACE inhibitors. METHODS: Fourteen patients with essential hypertension with night/day differences of mean blood pressure of more than 10% (dippers) aged 26 to 61 years (mean 52.9 +/- 9.2) and 14 healthy volunteers, 14 healthy volunteers were recruited and matched for age and gender. Ambulatory 24 hour arterial blood pressure measurement using HolCARD (Aspel, Poland) recorder with simultaneous ECG monitoring using the Mediarc-Premier IV Holter Monitoring System (DRG-International) undertaken in both groups of patients. The hypertensive patients were tested before and after one year of the enalapril treatment. The HRV analysis was performed according to a standard Fast Fourier Transformation (FFT). The time and spectral analysis parameters were compared within the examined groups of patients during day and night. RESULTS: The results obtained in the control group showed the great circadian fluctuations in sympathetic and parasympathetic activity. A simultaneous circadian HRV evaluation showed also significant increases in night RMSSD, TP, VLF, LF and HF mean values (p < 0.05) with significantly decreased night LF/HF ratio (p < 0.05) in comparison to the day time recordings. After one year of ACE inhibitors treatment, we noted the following circadian changes: Significant increase of RMSSD, pNN50 (p < 0.05), TP, VLF, LF and HF (p < 0.05) parameters with non significant increases in LF/HF ratio during night recording. A comparison between daily HRV parameters, before and after 1 year of ACEI treatment, presented significant increases in pNN50 and RMSSD mean values (p = 0.01) and decreases in LF/HF ratio, with simultaneous withdrawal of spectral parameters. However, no significant changes in comparison of night HRV parameters before and after ACEI intake were observed. CONCLUSIONS: Our observations support an attenuation of night vagal activity as an important factor in the pathogenesis of essential hypertension. The lack of effect of ACEI treatment on circadian heart rate variability parameters in patients with essential hypertension, despite long-term normalisation of blood pressure, suggests persistent circadian cardiac autonomic dysfunction in the "dippers" subgroup of patients. PMID- 12632904 TI - [Evaluation of self-measurement of blood pressure im ambulatory monitoring of hypertension therapy]. AB - OBJECTIVES: The aim of this study was to compare the usefulness and credibility of ambulatory self-measurement of blood pressure (SMBP) by hypertensive patients to automatic ambulatory blood pressure monitoring (ABPM). METHODS: The study was performed in 51 patients with primary hypertension (19 F, 32 M), aged 25-67 y divided into 3 groups: I-without treatment (n = 13), II-receiving monotherapy (n = 17) and III-treated at least with 2 hypotensive drugs (n = 21). For three consecutive days, patients measured their systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR) three times a day at 2 minute intervals during the following periods 06:00-08:00, 15:00-17:00 and 20:00-22:00. Afterwards, the process was repeated for three days using the automatic OMRON HEM 705C device. Results obtained on the second and third days were included in the calculation of SBP, DBP and HR mean values for the period 06:00-22:00. On the fourth day of the study, ABPM was performed with a Tycos Quiet Trak recorder. The parameters were measured every 20 minutes between 06:00 and 22:00. The average values for SBP, DBP and HR that were obtained with these two methods of blood pressure monitoring using the time period 06:00-22:00, were then compared. RESULTS: There was a significant correlation between SMBP and ABPM for SBP, DBP and HR in groups I, II and III (for SBP r = 0.81; 0.80; 0.82; for DBP r = 0.61; 0.62; 0.83; for HR r = 0.53; 0.69; 00.81). According to statistical tests, significant differences in the mean values of SBP, DBP and HR were found in the first (3.9 mmHg, 5.7 mmHg and 8 min-1 and second (4.9 mmHg, 6.2 mmHg and 6.2 min 1) groups. Only the HR (7.7 min-1) was statistically significant in the third group. CONCLUSIONS: Self-measurement of BP with automatic device is comparable to ABPM method of blood pressure monitoring, especially if multi-drug therapy is used for sustained hypertension. However, SMBP is not a completely alternative method for ABPM. PMID- 12632905 TI - Arterial hypertension in children treated at the Katowice Department of Pediatric Cardiology between 1993-2000. AB - In the literature, one can find a common opinion that 1-3% of the children population suffers from arterial hypertension. This rate depends on the diagnostic criterion and the studied population (age, sex, race etc.). The aim of this work was to present a rather large group (271 patients) treated for hypertension in our centre between 1993-2000. The ranged children from newborns to 18 year olds (mean = 13.75 +/- 4.5 years). In the study period, there is approximately a four fold annual increase in the number of hypertensive patients among all patients of the pediatric cardiology units. On the basis of our material, the rate of arterial hypertension depends on the age of the pediatric patients with as many as 3/4 of the children being teenagers > or = 14 years old. In the teenager group, all cases of essential hypertension were observed. Almost 70% of the studied children were boys. In about half the studied group, hypertensive changes on the eye fundus were found. One can suspect that automatic 24-hour measurement of arterial hypertension (so called "tension Holter") is of great value in children. A number of individual measurements exceeding the norm during "tension Holter" correlated with eye fundus vessel changes. In conclusion more attention should be paid to arterial hypertension in children. For example, in our previous paper published in 2000 it was stated that among 2.3% of "healthy" school children who suffered from arterial hypertension, non of these children were suspected of suffering from arterial hypertension. PMID- 12632906 TI - [Disturbances in microcirculation--an early predictor of arterial hypertension?]. AB - The aim of this study was to assess microcirculation and properties of large arteries in healthy adult subjects. In 28 individuals between the ages of 21 and 49, the microcirculation in the skin at resting conditions and under stimulation (hyperaemic reaction after a 3-minute long ischaemia at basal body temperature and at 44 degrees C) was measured with laser doppler flowmetry detection. Pulse Wave Velocity was measured by using Complior device and average values of blood pressure were calculated. Subjects were divided into two groups based on the median (pLDF = 374.5) of the hyperaemic reaction after ischaemia at basal body temperature (14 persons in each group). There were no significant differences in the age and pulse wave velocity in both groups. The group after ischaemia at basal body temperature was characterized by significantly lower values of the resting flow, higher values of blood pressure, and higher values of increase in flow after ischaemia when compared to the group after ischemia at 44 degrees C. Furthermore, there was a positive correlation between values in flow after ischaemia at basal body temperature and at 44 degrees C with systolic blood pressure (r = 0.79 and 0.62, respectively). Presented results suggest that disturbances in microcirculation can precede development of hypertension. PMID- 12632907 TI - [Vascular dysfunction in young patients with impaired glucose tolerance: insights from pulse wave velocity and endothelial function measurements]. AB - Endothelial dysfunction is regarded as an early marker of atherosclerosis and decreased arterial disstensibility has similar significance. The aim of this study was to evaluate endothelial function and arterial distensibility in patients with impaired glucose tolerance which may constitute markers of atherosclerosis. METHODS: Study group consisted of patients without chest pain, ischemic ECG changes, arterial hypertension or any risk factors for IHD, who were divided into two groups: Impaired glucose tolerance group (ITG)--48 patients with fasting glucose levels of 5.7 +/- 0.2 mmol/l and glucose levels of 9.2 +/- 0.6 mmol/l 2 hours after starting a 75 g oral glucose load and group 2-43 healthy age and sex matched controls with fasting glucose levels of 4.9 +/- 0.5 mmol/l and normal glucose tolerance. Endothelium dependent (EDV) and independent vasodilation (EIV) was assessed with 8 MHz high resolution ultrasound (Acuson Sequoia) according to Celermajer's method. PWV was measured with Complior Colson automatic device with measurement points above femoral and carotid artery. RESULTS: Flow associated with vasodilation was significantly impaired in ITG group (6.1 +/- 3.1% vs 13.4 +/- 3.9%-controls, p < 0.001). EIV was not significantly different between both groups. PWV was significantly higher in patients with impaired glucose tolerance than in controls (9.9 +/- 0.5 m/s vs 7.8 +/- 0.9 m/s) p < 0.001. CONCLUSIONS: Increased values of PWV indicate an increased arterial stiffness in patients with ITG, coexisting with endothelial dysfunction. These results reveal vascular dysfunction and potentially increased risk for development of atherosclerosis in ITG patients. PMID- 12632908 TI - [Comparison of quantitative echocardiographic methods with angiography for assessment of mitral regulation]. AB - The aim of this study was to compare echocardiographic assessment of the severity of mitral regurgitation (MR) with angiographic grading. MATERIAL: 51 patients (patients) with MR (43 men and 8 women), aged from 38 to 75 (mean 56 +/- 9 years), were used in the study. The etiology of MR was: Coronary artery disease (28 pts), infective endocarditis (3 patients), rheumatic disease (5 patients), dilated cardiomyopathy (7 patients), mitral valve prolapse (8 patients). EXCLUSION CRITERIA: Aortic stenosis and/or aortic insufficiency, mitral stenosis, mechanical prostheses and atrial fibrillation. METHODS: The mitral regurgitant volume (RV) and effective regurgitant orifice (ERO) were estimated by the proximal convergence method (PISA), regurgitant fraction (RF) and vena contracta width (VCW) by color Doppler quantitative method and compared to the cine ventriculographic grades. The angio-graphic severity of MR was classified as 1 to 4 grades according to the Sellers criteria. RESULTS: There was a good correlation between the angiographic grading and ERO (r = 0.855, p < 0.001), RV (r = 0.868, p < 0.001), RF (r = 0.923, p < 0.001) and VCW (r = 0.846, p < 0.001). CONCLUSIONS: 1. PISA method is clinically useful in routine evaluation of the severity of MR. 2. VCW provides a simple method for the identification of patients with severe MR. 3. These methods offer an alternative approach in quantifying MR noninvasively. PMID- 12632909 TI - [Seismocardiography--a noninvasive technique for estimating left ventricular function. Preliminary results]. AB - Seismocardiography (SCG) is a precardiac registration of low frequency waves generated during mechanical work of the heart and transmitted to the surface of the chest. It is a methodology derived from the field of seismology combined with signal analysis to evaluate the biomechanical function of the heart. SCG can be used for monitoring changes in cardiac function during stress testing. OBJECTIVES: Usefulness of SCG exercise testing for estimating left ventricular systolic and diastolic function in patients with coronary heart disease. The study population consisted of 50 patients divided into two groups: Group A with 20 patients after myocardial infraction aged 52 +/- 10 years with left ventricular dysfunction and ejection fraction min. 20%, max. 45%, mean 38%; Group B with 30 healthy volunteers aged 34 +/- 7 years. METHODS: All patients underwent symptom-limited exercise testing (ETT) combined with SCG testing. We measured ETT duration as well as workload, heart rate and blood pressure during ETT. In addition, we measured left ventricular systolic and diastolic time intervals (PEP, LVET, PEP/LVET, IRT) at rest and during SCG exercise testing. RESULTS: Group B exhibited better physical capacity; in particular the mean ETT duration was longer (9 +/- 2 vs. 7 +/- 2 min p < 0.0001), maximal workload was higher (168 +/- 28 vs. 118 +/- 33 W p < 0.0001), and heart rate during exercise was higher (162 +/- 10 vs. 129 +/- 25 p < 0.0001). In group A with lower EF PEP was prolonged (123 +/- 19 vs. 111 +/- 9 ms p < 0.05), PEP/LVET was higher (0.41 vs. 0.37) and IRT was increased (72 +/- 3 vs. 57 +/- 14) ms at rest and during exercise PEP (121 +/- 13 ms p < 0.05), PEP/LVET (0.39 vs. 0.37, IRT 51 +/- 25) vs. (38 +/- 16 ms p = 0.05). CONCLUSION: SCG may be helpful in monitoring left ventricular systolic and diastolic function in patients with IHD. PMID- 12632910 TI - [Clinical examination of Leber hereditary optic neuropathy in patients with the same gene mutation]. AB - Leber hereditary optic neuropathy (LHON), which is a relatively rare disease, is an inherited form of bilateral atrophy of optic nerves. This atrophy is related to degeneration of retinal ganglion cells and optic nerve axons. The aetiology of this disease is complex and involves primary mutations of mitochondrial DNA, as well as secondary genetic and/or epigenetic changes. We compared the clinical course of LHON in the patients with identical mitochondrial mutations. The molecular PCR-diagnosis was performed in 8 patients, representing 2 generations of the tested family. The group included 2 women suspected of being carriers and 6 men, 3 of whom were diagnosed with LHON and 3 suspected of being at risk of LHON. The primary 11,778 mtDNA mutation was diagnosed in all tested patients with one exception of the son of a male with LHON, which is in agreement with inheritance of mtDNA mutations. PMID- 12632911 TI - [Relationships between atopy and bronchial hyper-reactivity in Polish school age children]. AB - OBJECTIVE: To determine the prevalence of atopy and bronchial hyper-reactivity (BHR) and their relationship with wheezing and diagnosed asthma reported in schoolchildren. MATERIAL AND METHODS: In Cracow, 1347 schoolchildren (9.7 +/- 0.7 yrs of age) were surveyed with ISAAC questionnaire. Skin prick tests with 7 aeroallergens and the exercise challenge test for BHR evaluation were carried out. Atopy was established when a positive skin reaction with at least one allergen occurred. The BHR was confirmed in presence of at least 15% decrease of the relative post-exercise peak expiratory flow (D% PEF) or wheezing during or immediately after exercise. RESULTS: Atopy was found in 26.2% of schoolchildren, with a two-fold higher risk in boys/odds ratio (OR) = 2.14; 95% confidence interval (CI) (1.59-2.86)/. In the group of atopic children, wheeze in the last year and diagnosis of asthma occurred significantly more often. The risk of wheeze in last year related to the type of sensitizing allergen and was highest for Der. f./OR = 6.1; 95% CI (3.78-10.16)/, medium for allergens of trees, dog's fur, Der. p., cat's fur and grasses/OR = 5.17; 95% CI (2.27-11.76), OR = 4.36; 95% CI (2.35-8.13), OR = 4.00; 95% CI (3.12-8.01), OR = 3.86; 95% CI (2.18-6.82), OR = 2.58; 95% CI (1.50-4.45), respectively/. In case of Alternaria allergens no correlation with wheeze was observed/OR = 2.71; 95% CI (0.88-8.31)/. BHR was identified in 12.4% of schoolchildren and demonstrated a significant correlation with that reported in the questionnaire: wheeze in the last year/OR = 2.28; 95% CI (1.25-4.14)/, diagnosis of asthma/OR = 2.96; 95% CI (1.38-6.34)/and post exercise wheeze/OR = 3.17; 95% CI (1.60-6.27)/. Moreover, a significant correlation was demonstrated between atopy and BHR/OR = 1.85; 95% CI (1.22 2.79)/. In the group of children without wheeze in the past and without diagnosis of asthma, BHR was significantly higher in atopic children (positive skin test to in-door allergens) comparing to non-atopic ones, mean D% PEF values were 3.95%, 0.39%, respectively (p. = 0.0014). CONCLUSIONS: In children, there is a significant correlation between atopy and BHR, modified by individual history of wheeze or asthma, sex and allergen type. PMID- 12632912 TI - [Psychological consequences of moderate to severe cranio-cerebral injuries: dysexecutive syndrome. Preliminary report]. AB - The late sequel of cranio-cerebral trauma are common causes of decreased mental capability and pathology localized in frontal lobes which can cause the dysexecutive syndrome (the individual has a goal but has difficulty in generating or activating the appropriate goal directed behavior). We analysed 58 patients, random-chosen, treated in the Neurotraumatology Department in Krakow after craniocerebral injury. Among them, 6 patients were found who scored more than average in DEX questionnaire. It was shown that diagnosis of DEX enables use of neuropsychological rehabilitation, which can improve final outcome. PMID- 12632913 TI - [Influence of tobacco dust on the respiratory system and selected immunological parameters]. AB - Tobacco dust contains various immunologically active as well as toxic substances. However the relationship between allergic reactivity or lung function with chronic exposure to tobacco dust remains unclear. Accordingly the aim of the present study was to investigate the relationship between occupational chronic exposition to tobacco dust, respiratory function and some allergic reaction parameters. METHODS: 40 tobacco factory workers (47.5% women and 52.5% men) aged 25-59 years (mean 36.5) chronically (5-31 years, mean 12.6) exposed to tobacco dust were included into the study. Control group consisted of 30 subjects (46.7% women, 53.3% men) aged 25-60 years (mean 36.6) not exposed to tobacco leaves' dust. Detailed epidemiological data was collected. Additionally total IgE, specific (tobacco) IgE, eosinophil blood counts, skin tests (mixed grass and weed pollens, house dust, feather, tobacco extracts), basophil degranulation and neutrophil destruction tests with tobacco extracts as well as spirometry were studied in these groups. We found that FEV1/VC was significantly lower in tobacco industry workers chronically exposed to tobacco dust than in the control group (91.5 +/- 11.6% vs. 101.7 +/- 10.7% n; p = 0.0004). These subjects were also characterized by higher occurrence of mild bronchial obturation (FEV1/VC < 88% and FEV1 > 70%) which was present in 30% tobacco factory workers and in 6.7% of control group (p = 0.035). Levels of total IgE and tobacco-specific IgE, eosinophil counts, skin test reactivity, basophil degranulation and neutrophil destruction tests were not different between groups. CONCLUSIONS: Occupational chronic exposition to the dust of tobacco leaves is associated with significant increase in the occurrence of mild obturative ventilatory disturbances. Simultaneously no increased frequency of allergization to tobacco or other allergens was observed in tobacco industry workers. PMID- 12632914 TI - [Role of procalcitonin in the diagnosis of uncomplicated infective endocarditis]. AB - Infective endocarditis especially where blood culture is negative frequently causes problems in diagnosis despite of many nonspecific inflammation parameters. Procalcitonin (PCT) concentration is a new marker of severe bacterial and fungal infection. The aim of this study was to evaluate the usefulness of PCT concentration assessment in the diagnosis of uncomplicated infective endocarditis of bacterial etiology. The study group consisted of 30 patients with ongoing infective endocarditis in the course of acquired valvular heart disease. The diagnosis of bacterial endocarditis was established according to the Duke criteria on the basis of: clinical examination, laboratory investigations (inflammatory parameters, transthoracic and transesophageal echocardiography) and positive blood cultures. Patients with sepsis, concomitant infections and in an early postoperative period were excluded. Two control groups consisted of: 10 patients without endocarditis and other infections and another 10 patients without endocarditis and with an urinary tract infection. Serum procalcitonin concentrations were measured by an immunoluminometric assay (LUMItest PCT set). Mean serum PCT concentrations in patients with endocarditis (0.12 +/- 0.13, range 0-0.4 ng/ml) were significantly higher than in control group without infection (0.03 +/- 0.05, range 0-0.1 ng/ml) and higher than in control group with an urinary tract infection (0.02 +/- 0.04, range 0-0.1 ng/ml). However in 12 patients (of 30) were below sensitivity of the method and in the rest were within normal range (< 0.5 ng/ml). CONCLUSION: Serum PCT concentration assessment seems to have no value in the diagnosis of uncomplicated infective endocarditis. PMID- 12632915 TI - [Acute normovolemic hemodilution together with aprotinin applied as a scheme of blood saving in cardiac surgery]. AB - The risk of blood loss in open heart surgery procedures is related to a high level of homologous blood transfusion. Due to numerous possible complications connected with transfusion, as well as the increase in prices of blood preparations, methods of blood saving are of great interest. The aim of the study was to assess acute normovolemic hemo-dilution (ANH) efficacy and aprotinin administration in homologous blood usage limitations. The study was conducted in the group of 265 patients operated on during the period of 12 months. The control group consisted of patients operated on during the period 6 months. The scheme of ANH with the administration of aprotinin was applied in the second half-year period. A statistically significant reduction in the percentage of patients who required blood transfusions was observed (77.2%/53%). The scheme applied enabled to reduce procedure costs in regard to blood preparation purchase. The results have shown that ANH together with antifibrinolytic drug administration is not only an efficient but also profitable strategy of blood saving. PMID- 12632916 TI - [Comparison of early clinical results after left ventricular aneurysmectomy using classic and modified technique]. AB - INTRODUCTION: In our department left ventricular aneurysmectomy was performed using only classic Cooley technique until 1989. Since then a modified technique was introduced: left ventricular geometry reconstruction modo Stoney simultaneously with the excision of scarred endocardium. AIM: Comparison of early clinical results after left ventricular aneurysmectomy using classic (Group A) and modified technique (Group B). MATERIAL AND METHODS: Retrospective data analysis of early clinical outcome of patients (pts) operated from 1989 to 2000: classic technique 74 pts [63M (85.1%); 11F (14.9%); average age 52.26 +/- 10.35 years], modified technique 86 pts [74M (86%); 12F (14%); average age 55.83 +/- 9.3]. RESULTS: No statistically significant differences between groups considering number of critically stenosed coronary arteries, left ventricular ejection fraction and left ventricular diameters were observed. Mean number of bypass grafts (gr. A 1.55, gr. B 1.69); aortic crossclamping time (gr. A 48.1 (24 77) +/- 12.9; gr. B 50.9 (0-91) +/- 16.1) were not significantly different between groups. In gr. A, 11 (14.8%) pts died: low output syndrome (LOS) 3 pts, LOS and ventricular dysrhythmias (VD) 5 pts, VD 3 pts. In gr. B 4 (4.6%) pts died: LOS 3 pts; LOS and VD 1 pt. Among other complications LOS and VD occurred significantly more frequently in gr. A. In both groups average postoperative ejection fraction increased; twice higher in gr. B than A. CONCLUSIONS: Modified technique: 1. Significantly decreases incidence of ventricular dysrhythmias in comparison to classic technique. 2. Significantly lowers incidence of low output syndrome and mortality rates in comparison to classic technique. PMID- 12632917 TI - [Is autonomic neuropathy a risk factor of death in dialysed patients?]. AB - The aim of the study was to determine, whether autonomic neuropathy is a risk factor of death in dialysed patients. Results of autonomic tests (R-R interval variation and sympathetic skin response) which were performed 3 years ago in 51 patients were analysed. 22 of these patients died. A greater incidence of dysautonomia was found in patients who died. However, due to the small number of examined patients, it cannot be concluded, that dysautonomia is a risk factor of death in uremia. PMID- 12632918 TI - [Psychological characteristics of children suffering from Duchenne muscular dystrophy]. AB - The authors examined intellectual and socio-emotional functioning of 39 subjects suffering from Duchenne muscular dystrophy. Their school situation, access to rehabilitation and a quality of familial upbringing attitude were characterized, as well. No significant differences concerning I.Q. between sick children and healthy population were found. PMID- 12632919 TI - [Pregnancy induced arterial hypertension--an attempt to evaluate hermetic aspects of the blood-brain barrier connected with skull computed tomography]. AB - The aim of the work was to study the influence of pregnancy induced arterial hypertension (PIH) on hermetic aspects of the blood-brain barrier. Neurological consultation was followed by cranial computed tomography which revealed changes within structures of central nervous system. The observations were performed in 6 pregnant women, assigned to control group--C, and study group--G. All women had caesarean section performed in conductive anaesthesia. The method included collection of 15 ml venous blood and 4 ml cerebrospinal fluid (CSF). Albumin and immunoglobulin G (IgG) serum concentrations, micro-albumin and IgG CSF concentrations, and permeability indexes for albumin and IgG--Qalb and QIqG, were estimated. In contrast to group C, increased Qalb and QIgG indexes proved lack of integrity of blood-brain barrier in the group G. Control cranial CT scans revealed partial subsidence of changes after 3-5 days, and complete subsidence after 9-14 days. PMID- 12632920 TI - [The phagocytic activity of blood platelets in different parasitic diseases]. AB - Blood platelets take part in immune reactions of the organism, especially in anti parasitic immunity. We conducted the phagocytic activity of blood platelets. This experiments were done on 40 patients infected with Giardia intestinalis (26), Ascaris lumbricoides (8), Entamoeba histolytica/Entamoeba dispar (6). Blood for analysis was collected thrice, before treatment (P1), after two weeks (P2), and after two months treatment with anti-parasitic drugs (P3). Control group consisted of 32 healthy people. The evaluation of platelet phagocytic activity revealed that the mean percentage of phagocytizing platelets in patients was 2.87 (P1), 2.94 (P2), 2.86 (P3), being significantly lower in controls (mean 2.15), the difference was statistically significant (p < 0.05). No statistically significant difference was found in the mean phagocytic index between patients (P1, P2, P3) and controls. Our general conclusion is--parasites are able to stimulate phagocytic activity of blood platelets. PMID- 12632921 TI - [Influence of hemodialysis on expression of glycoprotein lb platelets reactivity in patients with the end stage renal failure]. AB - Complex disturbances in hemostasis characterise chronic renal insufficiency. Defect of primary hemostasis is a common cause of bleeding complications. The hemodialysis procedure (HD), by itself, influences platelet function (the key part of primary hemostasis) which is compromised after the procedure. Many functional defects of thrombocytes have been described in end stage renal failure (ESRF) patients; one of them is adhesion defect where the pivotal role plays the complex of glycoprotein Ib (GP Ib) and IX. The aim of the study was to determine the extent of activation, reactivity of platelets and expression of GP Ib in ESRF patients. The flow cytometry method was used, and thrombin was used for stimulation of thrombocytes. Membrane expression of GP Ib and selectin P was assessed in 14 hemodialysed patients before and after stimulation with thrombin, before and after HD and in 10 healthy persons. The patient group had lower expression of GP Ib on resting platelets and significantly lower expression of selectin P after stimulation with thrombin when compared to the control group. After HD, thrombocytes had much lower expression of GP Ib; however there was no difference in expression of selectin P when compared to the state before HD. A lower reduction of GP Ib expression after stimulation with thrombin was observed after and before HD. On the basis of the results the following conclusions may be drawn: hemo-dialysed ESRF patients have lower expression of platelet GP Ib and reactivity of thrombocytes; the HD results in further depression in expression of GP Ib and reactivity of thrombocytes. It is plausible that nitric oxide (NO) released during HD modulates the expression of selectin P, but it remains to be confirmed. PMID- 12632922 TI - [Structure and function of PDGF-R-alpha and its expression in normal kidney and kidney diseases]. AB - Platelet-derived growth factor is commonly known as a mitogen. Many research data suggest a role for PDGF-beta R in the mitogenic response of mesangial cells. There are four members of PDGF family known as PDGF-A chain, PDGF-B chain, PDGF-C chain and PDGF-D chain, which in active forms are dimers. As far as two receptors PDGF-alpha R and PDGF-beta R are known to bind PDGF. There is a difference in binding affinity of various forms of PDGF by these receptors. Two different promotors P1 and P2 can be used for PDGF-alpha R gene transcription. There are several different haplotypes of promotor P1 sequence. Transcription of PDGF-alpha R gene is under control of many factors. Interaction between a receptor and its ligand includes receptor dimerisation and autophosphorylation of tyrosine residues. PDGF AA is unique in that it can only be bound by alpha-receptor dimer. PDGF-AA expression has been confirmed in the normal kidney, as well as in several renal diseases. Although the expression of PDGF-alpha R has been found to accompany that of PDGF-AA, its actual relevance for the development of the glomerular pathology is not clear. PMID- 12632923 TI - [Neural areas of emotions in normal and brain pathology assessed by functional neuroimaging methods]. AB - The investigation into neural back-ground of emotions in normal and brain pathology using functional brain imaging techniques (fMRI, PET, SPECT) remains one of the most interesting scientific challenges in the area of psychiatry. The aim of the current study is: 1) to present the recent neuroimaging findings concerning functioning of "emotional brain" in normal and brain pathology with particular emphasis on psychiatric disorders, 2) to consolidate knowledge of neuroimaging techniques and experimental designs used in the assessment of human affect. PMID- 12632924 TI - [Validation of organ failure scoring systems in objective illness characterization]. AB - Multiple organ failure syndrome is the leading cause of death in intensive care units. Medical scales and scoring systems are used for objective characterization of patients, quantitation of illness severity, prognosis, identification of high risk patients, determination of treatment effectiveness and final outcome. We reviewed some data about the most frequently used scoring systems describing organ failure. Multiple Organ Dysfunction score (MODs) grades increasing severity of dysfunction of vital significant organ systems and it reflects intensity of SIRS.APACHE II and APACHE III scoring systems and SOFA score are used widely in clinical practice. PMID- 12632925 TI - [Sudden cardiac death in children]. AB - Natural, sudden deaths in children are the cause of about 5% of all deaths. The most frequent cause appears to be pathology of the cardiovascular system. Direct cause of death are arrhythmias or, rarely, haemodynamic disturbance coexisting with decrease in minute cardiac output. Authors present the most frequent diseases (myocarditis, hypertrophic cardiomyopathy, long QT syndrome and preexcitations syndromes) as well as congenital heart defects (aortic stenosis, tetralogy of Fallot, transposition of the great arteries, Ebstein's syndrome, anomalies of coronary arteries) which can be causes of sudden death in childhood. PMID- 12632926 TI - [Psoriasis and cardio-vascular disorders]. AB - Psoriasis is one of the most frequent skin diseases. It occurs in adult people with a frequency 2-3%. Psoriasis is a systemic disease with immunologic and genetic determinants. Micro and macro vascular disorders are important for pathogenesis of psoriasis. Authors attempt to show a current scientific statement of pathogenetic mechanisms in psoriasis with special consideration of factors involving cardiovascular abnormalities. In consecutive steps described are associations between psoriasis and arterial hypertension from etiological, pathological and epidemiological point of view. In the same order are reviewed the junctions of psoriasis and lipid and carbohydrate metabolism disorders, as well as coagulative and heart valves abnormalities. PMID- 12632927 TI - [Influence of ethanol on oxidative stress in the liver]. AB - Ethanol ingestion causes an increase in free radical generation in the liver mainly by induction of microsomal cytochrome P-450, conversion of xanthine dehydrogenase into xanthine oxidase in cytosol and increased one electron oxygen reduction in mitochondria. As a result, the decrease in antioxidant status characterized by changes in activity of antioxidant enzymes and by decrease in nonenzymatic antioxidant level with different intensification depended on acute or chronic alcohol ingestion. The consequence of the above changes is oxidative stress in the liver. It intensifies free radical action on cell compartments which are characterized by oxidative modification of lipids, proteins and nucleic acids. These cytotoxic processes may finally lead to alcoholic liver injury in the end. PMID- 12632929 TI - [Diagnosis and treatment of thyroid nodules]. AB - The paper presents current diagnostic and therapeutic approach of thyroid nodules. TSH and iodine are the main goitrogenic agents, but there are also other auto- para-endocrinal factors that play very important roles in goiter growth as well as in the intrinsic growth potential of some thyroid follicular cells. The diagnosis of thyroid nodule demands further investigation such as thyroid function evaluation and distinction between benign from malignant nodules using FNAB. The confirmation or suspicion of malignancy or presence of compressive symptoms is indication for surgery. In benign lesions, without features of hyperthyroidism observation may be advised. Thyroxine therapy may be proposed to some patients but the possibility of side-effects should always be taken into account, as well as the low efficacy of this treatment. PMID- 12632928 TI - [Mechanism of multidrug resistance of ovarian cancer]. AB - Ovarian cancer is one of the most frequent cancers in women, and five-year survival rate from ovarian cancer is only 20% to 30%. The main problem and reason for lack of success in treatment is usually late recognition. Unfortunately, advances in surgical techniques and chemotherapy do not translate into improved survival. Despite the high response rates with initial treatment, most patients with ovarian cancer relapse. Usually then they are chemoresistant to standard chemotherapy. This article presents a review of the basic mechanisms of drug resistance in ovarian cancer and their clinical significance. PMID- 12632930 TI - [Application of research results of tissue engineering in wound healing]. AB - Based on current literature wound healing and repair have been analyzed. Basic types of dressing materials used in wound repair as well as the use of skin grafts for covering skin lesions have been discussed. Taking into account the results of research in the fields of molecular biology and tissue engineering the authors describe the possibilities of producing skin substitutes, transplanting in vitro-cultured epithelial cells and gene therapy of various skin diseases and systemic diseases. PMID- 12632931 TI - [False aneurysms after button-Bentall procedure]. AB - Between January 1995 and December 2000, 222 consecutive patients underwent complete aortic root replacement in the Department of Cardiovascular Surgery of the Hertz- und Gefaess- Klinik- Bad Neustadt (Germany). At the exception of 2 cases, all patients received the modified Bentallde Bono procedure with total excision of the ascending aorta and direct reimplantation of the coronary ostia using button technique. During the follow-up period of up to 6 years in 3 patients, a periprosthetic aneurysm was diagnosed. In two patients with clinical symptoms and a fistula between the prosthetic lumen and the spurium aneurysm, a reoperation was performed. Due to the complete absence of clinical symptoms or a fistula connecting the prosthesis and the aneurysm or progress in size of the aneurysm during the follow-up period the third patient was treated conservatively. The choice of proper treatment of this complication is difficult and should be based on continuous investigation of clinical symptoms and various technical examinations. PMID- 12632932 TI - [Recombinant erythropoietins--an alternative therapy to red cell blood transfusions in infants with hereditary spherocytosis]. AB - Hereditary spherocytosis is the most frequently occurring haemolytic anaemia. Some patients manifest clinical signs during the neonatal period and require transfusion. A case of hereditary spherocytosis in neonate is presented. The use of rhEPO during the first months may be an alternative therapy to red cell transfusion. PMID- 12632934 TI - Gristle in the sausage.... PMID- 12632933 TI - [Laparoscopy in abdominal emergencies]. AB - Up-to-dated concepts about the use of laparoscopy in the "acute abdomen" have been discussed. Most frequently performed procedures in abdominal emergencies and trauma were presented. Indications, contraindications, advantages and drawbacks with regard to own experience and literature were discussed. PMID- 12632935 TI - Ballistic fingerprinting databases. AB - This article discusses a number of questions regarding the setting up of ballistic fingerprinting databases, consisting of ammunition components fired by all the firearms held in legal possession. These questions can be classified into three categories--the efficiency of the database, forensic issues, and practical issues to be dealt with. The current New York State legislation is used as an illustration of the choices to be made when setting up a ballistic fingerprinting database. Three important arguments are formulated against the installation of a ballistic fingerprinting database. PMID- 12632936 TI - The collation of forensic DNA case data into a multi-dimensional intelligence database. AB - The primary aim of any DNA Database is to link individuals to unsolved offenses and unsolved offenses to each other via DNA profiling. This aim has been successfully realised during the operation of the New Zealand (NZ) DNA Databank over the past five years. The DNA Intelligence Project (DIP), a collaborative project involving NZ forensic and law enforcement agencies, interrogated the forensic case data held on the NZ DNA databank and collated it into a functional intelligence database. This database has been used to identify significant trends which direct Police and forensic personnel towards the most appropriate use of DNA technology. Intelligence is being provided in areas such as the level of usage of DNA techniques in criminal investigation, the relative success of crime scene samples and the geographical distribution of crimes. The DIP has broadened the dimensions of the information offered through the NZ DNA Databank and has furthered the understanding and investigative capability of both Police and forensic scientists. The outcomes of this research fit soundly with the current policies of 'intelligence led policing', which are being adopted by Police jurisdictions locally and overseas. PMID- 12632938 TI - 4-Bromo-2,5-dimethoxyphenethylamine (2C-B): a review of the public domain literature. PMID- 12632941 TI - Police surgeons--training for quality. PMID- 12632943 TI - A professional body for forensic science. PMID- 12632942 TI - 'Agony and ecstasy': drug abuse from laboratory to court, clinic and graveyard. PMID- 12632965 TI - Incidental detection of hepatitis B surface antigen: a growing concern for the Indian physician. PMID- 12632966 TI - Management of hepatitis B and C in renal failure and renal transplant recipients. PMID- 12632967 TI - Management of ingested foreign bodies: a review. PMID- 12632969 TI - Abdominal aortic aneurysm and peptic ulcer disease: experience of a single unit in Sri Lanka. PMID- 12632968 TI - Peritoneoscopy: experience in Sudan. AB - INTRODUCTION: The use of peritoneoscopy is an established method for diagnosis of intra abdominal diseases. In industrialized countries such as USA and Germany its use has declined with the advent of modern imaging techniques. However it is believed that there is an important rationale for its use in developing countries. STUDY AIMS: A retrospective study to assess the value of peritoneoscopy in a developing country (Sudan). PATIENTS AND METHODS: A total of 274 patients were included in this study. Peritoneoscopy was performed in the endoscopy theatre, using standard techniques. Biopsy specimens were taken, when required, from relevant tissues. The patients were closely monitored during and following the procedure. RESULTS: Peritoneoscopy was indicated for assessment of hepatomegaly in 108 (39.1%) patients, exudative ascites 66 (24%), diagnosis of abdominal mass 48 (17.5%) and miscellaneous indications 52 (19.4%). The final diagnosis of all cases after peritoneoscopy included malignancies other than hepatocellular carcinoma (HCC) 81 (29.8%) patients, miscellaneous conditions 60 (21.8%), HCC 45 (16.4%), liver cirrhosis 26 (9.4%), abdominal tuberculosis 24 (9.4%), normal liver 16 (5.8%), hepatosplenic schistosomiasis (HSS) 15 (5.5%) and amoebic liver abscess 7 (2.5%). The peritoneoscopic appearance of HSS was very characteristic. CONCLUSION: This large study confirms that peritoneoscopy has a high diagnostic yield and is a cost-effective technique for use in developing countries. PMID- 12632970 TI - High prevalence of hepatitis B virus infection amongst Tibetans in Nepal. AB - Tibetans have been living in Nepal since 1959. Study of the prevalence of viral hepatitis among them showed that they have a high prevalence of hepatitis B virus (HBV) infection. Prevalence of total HBV infection and Hepatitis B surface antigen (HBsAg) among them was 61% and 16% compared to 10.0% and 0.7% respectively among the Nepalese. The predominant HBsAg subtype among the Tibetans was 'ayw'. Perinatal and childhood transmission was found important in the spread of HBV infection among the Tibetans. PMID- 12632971 TI - Functional outcome and patient satisfaction after ileal pouch anal anastomosis for ulcerative colitis in a developing country. AB - AIM: To study the long-term outcome and patient satisfaction of patients with an ileal pouch-anal anastomosis (IPAA) for ulcerative colitis in India. PATIENTS AND METHODS: We studied 35 patients who had undergone IPAA for ulcerative colitis between 1985 and 1998 and had intestinal continuity restored for more than 6 months. These patients were asked to answer a detailed questionnaire on their bowel function, urogenital function, etc. A complete haemogram, serum iron studies, liver function tests and D-Xylose absorption test were done. In addition hepatobiliary ultrasound, stool microscopy, pouchoscopy and pouch biopsies were also performed. Patient satisfaction after the procedure was also evaluated. RESULTS: Thirty-five patients (17 men and 18 women) underwent a complete evaluation. The duration after restoration of continuity ranged from 6 months to 164 months (mean 78.6 months). The mean stool frequency was 7.2 stools per 24 hours. Five patients had urgency of stool, 9 had occasional soiling and 1 had major incontinence. Four patients had minimal restriction of social activities and 1 discontinued his employment. All patients were sexually satisfied except one man who had impotence and one woman who had dyspareunia. Fifteen patients had abnormal serum iron studies (Haemoglobin < 9 g/dl in 11). Eleven patients had D Xylose absorption below normal values. Two patients were found to have gallstones. All pouch biopsies showed chronic inflammation and 1 patient had histological evidence of pouchitis. Eighty-five percent of patients reported that they were very satisfied with the procedure. CONCLUSION: Good functional recovery and acceptance of the procedure over the long term suggests that it is a valid procedure to be recommended for patients with ulcerative colitis in India. PMID- 12632972 TI - Falciparum hepatopathy: a reversible and transient involvement of liver in falciparum malaria. AB - Between August 1992 and May 1997 a total of 56 patients with falciparum malaria and liver disease were seen. The serum bilirubin, mainly of conjugated variety was raised. The ALT was less than 3 times normal with a minimally raised or normal Alkaline Phosphatase (ALP) and prothombin time. The liver function abnormalities were not related to grade of parasitaemia, fever, duration of the illness, nutritional status of the patient or associated medical problems. Treatment of falciparum malaria led to quick reversal of liver function tests. PMID- 12632973 TI - Bacterial profile of dairy products sold in Chandigarh. AB - Gastrointestinal infections are known to occur due to bacterial contamination of dairy products. A total of 141 dairy products viz. kulfi, ice cream and softy samples were investigated bacteriologically. Staphylococcus was the predominant organism isolated followed by Klebsiella pneumoniae, Escherichia coli and Campylobacter. Two of the samples also yielded Yersinia. The total bacterial counts obtained ranged from 2 x 10(1) to 5.2 x 10(11) colony forming units per ml (CFU/ml) for kulfi, 4 x 10(1) to 9 x 10(9) CFU/ml for ice cream and 2 x 10(1) to 2 x 10(10) CFU/ml for softy samples. The high degree of bacterial contamination seen indicates poor hygienic conditions and faults in manufacturing/handling of dairy products during and after processing and production. PMID- 12632974 TI - Epidemiology of hepatitis E and A in Ludhiana. AB - AIM: To study the epidemiology of hepatitis E and A in Ludhiana. METHODS: A retrospective analysis of laboratory data of the year 2001 was done, to study the epidemiology of hepatitis E and A in Ludhiana in relation to prevalence, age, sex, area of residence and seasonal variation. RESULTS: Prevalence of HEV was 13.9%. Infection with it occurred through out the year with no preponderance of cases following the monsoons. Young male adults between 20-40 years of age were predominantly affected. Prevalence of HAV was 5.5% affecting mainly children below 14 years of age. Population of urban areas around "Gandha Naliash" (a large open sewer) was at high risk as there was clustering of HEV & HAV cases around that area. PMID- 12632975 TI - Helicobacter pylori in Indian HIV infected patients. AB - Helicobacter pylori is a causative organism for chronic gastritis and associated with peptic ulcer disease. Infection may be asymptomatic as well. Human immuno deficiency virus infection predisposes to a multitude of opportunistic infections, many of them resulting in gastrointestinal symptoms. We studied the prevalence of H pylori co-infection with HIV and its correlation with gastrointestinal symptoms in HIV infected patients. Seventy-three consecutive HIV infected patients presenting to the medical out patient department of Postgraduate Institute of Medical Education & Research, Chandigarh, India, were included in the study. Antibodies (IgG) to H pylori were tested by ELISA. There were 43 males, 30 females; mean age 26.1 +/- 4.7 years. Risk factors for acquiring HIV infection was predominantly heterosexual exposure. Eleven patients presented with gastrointestinal symptoms. Thirty-five of the 73 (47.9%) patients had serological evidence of H pylori infection. Six of them had gastrointestinal symptoms. These were odynophagia in 5, dyspepsia in 4 and recent diarrhoea in 2. Twenty-four patients with H pylori infection had AIDS. There was no difference in the prevalence of H pylori infection between patients with and without AIDS. PMID- 12632976 TI - Protective effect of natural honey against acetic acid-induced colitis in rats. AB - AIMS: The protective effects of natural honey against acetic acid-induced colitis were investigated in rats. METHODS: Honey and glucose, fructose, sucrose, maltose mixture were administered, orally and rectally, daily for a period of 4 days. Induction of colitis was done on the third day using 3% acetic acid. Animals were killed on day 4 two hours after administration of the dose and colonic biopsies were taken for macroscopic scoring, histopathological and biochemical studies. RESULTS: Honey dose-dependently afforded protection against acetic acid-induced colonic damage. There was almost 100% protection with the highest dose (5 g/kg) used while glucose, fructose, sucrose, maltose mixture produced no significant protective effect. Also, honey prevented the depletion of the antioxidant enzymes reduced glutathione and catalase and restored the lipid peroxide malondialdehyde towards normal levels. CONCLUSIONS: Further studies are required to explore the active ingredients responsible for the antioxidant effect of honey and its therapeutic potential in humans. PMID- 12632977 TI - Exacerbation of ulcerative colitis by cytomegalovirus infection in an immunocompetent Indian patient. AB - A patient with typical features of idiopathic ulcerative colitis, in remission, developed an attack of severe colitis. Sigmoidoscopy showed submucosal black nodules in the sigmoid colon. Mucosal biopsies from the involved areas showed evidence of acute on chronic colitis with cytomegalic cells and intra-nuclear inclusions suggestive of cytomegalovirus (CMV) disease. The patient attained remission following subtotal colectomy and intravenous ganciclovir therapy for 3 weeks. The patient had another relapse five months later. The colonic biopsies during this relapse showed evidence only of idiopathic ulcerative colitis, with no CMV infection. The patient responded well to steroid therapy. PMID- 12632978 TI - Histoplasmosis of the liver: a rare case. AB - A 39 year old male presented with history of fever and jaundice for 3 months. A liver biopsy showed numerous ovoid fungal bodies around 5 mm in size in the macrophages and Kupffer cells. A diagnosis of hepatic histoplasmosis was made which is an uncommon entity in our country. PMID- 12632979 TI - Recurrent biliary cystadenoma. PMID- 12632980 TI - Neoadjuvant chemotherapy in advanced gastric cancer--results of a pilot study. AB - INTRODUCTION: Adenocarcinoma of the stomach is usually advanced at presentation, and local or distant spread may preclude the option of primary curative resection. Neoadjuvant chemotherapy (NAC) has shown promise in downstaging initially unresectable disease. This pilot study was planned to assess the utility of NAC using Cisplatin and 5-Fluoro uracil in the management of initially unresectable gastric cancer. PATIENTS AND METHODS: Ten patients with unresectable gastric adnocarcinoma were included. They received two cycles of cisplatin, 30 mg/m2 intravenously in combination with 5-Fluoro Uracil, 1000 mg/m2. They were restaged using Endoscopy and CT scan and taken up for exploratory laparotomy. RESULTS: Eight of 10 patients (80%) had an objective response to chemotherapy. Six patients (60%) with initially unresectable disease could be offered curative surgery. The median survival was 10 months (range 1-60 months). There were two long term survivors (48 and 60 months respectively). CONCLUSION: Neoadjuvant chemotherapy (NAC) is an effective option in downstaging initially unresectable gastric carcinoma. Complete response to chemotherapy also predicts long term survival. PMID- 12632981 TI - Experience of surgical management of pseudo-aneurysms of branches of the coeliac axis in a North Indian Hospital. AB - BACKGROUND: Bleeding splanchnic artery pseudo-aneurysm is a rare but frequently fatal complication that can be successfully managed by angiographic embolization. However, certain patients because of hemodynamic instability, non-availability of technique or angiographic failure may require primary surgical intervention. METHOD: Retrospective review of 13 patients presenting with exsanguinating hemorrhage from ruptured pseudo-aneurysm arising from branches of coeliac axis, managed surgically in absence of angiographic embolization. RESULTS: Splenic artery was most commonly involved (n = 7) followed by hepatic (n = 3), gastroduodenal (n = 2) and left gastric artery (n = 1). The most common underlying aetiology was pancreatitis (n = 8, acute = 2; chronic = 6) followed by iatrogenic (n = 3), liver abscess (n = 1) and gastric ulcer (n = 1). Seven patients presented with upper gastrointestinal (GI) bleed, while 2 each with lower GI bleed, haemobilia and bleeding through tube drains. CT-scan accurately demonstrated the pseudo-aneurysm in 11 (84.6%) patients and additionally demonstrated the underlying pathology. The surgical management chiefly consisted of ligation of offending vessel and additional procedures directed at primary pathology. Overall, 77% patients had a favourable outcome while 23% died consequent to ongoing hemorrhage. CONCLUSION: Pseudo-aneurysm involving the branches of coeliac axis most commonly arises as a result of pancreatitis and affects splenic artery. CT-scan accurately demonstrates pseudo-aneurysm and associated pathology in majority of cases. Primary surgical management in the presence of hemodynamic instability and non-availability of angiographic embolization is a viable alternative. PMID- 12632982 TI - Pseudoaneurysm in the gallbladder fossa with recurrent ruptures. PMID- 12632983 TI - Polyvisceral gangrene due to radiation enteritis. PMID- 12632984 TI - Retroperitoneal Ewing's Sarcoma: a case report. PMID- 12632985 TI - Endoscopic sclerotherapy for bleeding oesophageal varices: experience in Sudan. AB - INTRODUCTION: Bleeding due to oesophageal varices is the commonest cause of upper gastrointestinal tract haemorrhage in Sudan. Endoscopic injection sclerotherapy (EST) is a valuable therapeutic modality for the management of variceal bleeding. Other options for treatment such as variceal banding are either expensive or unavailable. STUDY AIMS: A retrospective study to evaluate the outcome of EST in the management of bleeding oesophageal varices due to portal hypertension in a developing country (Sudan). PATIENTS AND METHODS: A total of 1070 patients over a period of 10 years (1986-1996) were studied. Inclusion criteria was bleeding oesophageal varices consequent to portal hypertension. EST was performed using a standard technique. Ethanolamine oleate 5% was the sclerosing agent utilized. The procedure was done on a day- case basis. RESULTS: There were 904 males (84.5%) and 166 females (15.5%). The cause of portal hypertension was schistosomal periportal fibrosis (PPF) in 999 (93.3%) patients, liver cirrhosis 59 (5.5%), mixed PPF and cirrhosis 5 (0.46%), portal vein thrombosis 6 (0.64%) and congenital hepatic fibrosis was present in 1 patient. A total of 100 (9.4%) patients presented with bleeding which occurred after surgery. Full obliteration of varices required a mean of 4 sessions with a range of 2-6. 462 (43.2%) have been followed up until complete sclerosis of varices. CONCLUSION: This study provides evidence that endoscopic injection sclerotherapy is an essential component in the management of bleeding oesophageal varices caused by portal hypertension. It is a feasible and a cost-effective therapeutic strategy in developing countries. PMID- 12632986 TI - The Medical Society of New Jersey's journal. PMID- 12632987 TI - Health care providers' responsibilities toward hearing-impaired patients. PMID- 12632988 TI - Recommendations to reduce the risk of occupational HIV transmission. PMID- 12632989 TI - Referring to the report from the Office of the Inspector General. PMID- 12632990 TI - New Jersey's pharmaceutical industry. PMID- 12632991 TI - [Epidemiology of scooter accidents in Italy: the effectiveness of mandatory use of helmets in preventing incidence and severity of head trauma]. AB - The article refers to two surveillance studies, conducted the first in the city of Rome and Naples, in 1999, and the second only in Rome before and after the implementation of the new law on mandatory use of helmet (1999-2000). Personal data, accident circumstances, helmet use, others involved in accident and health data were collected. In the first study we found that only 12% of injured people wear a helmet. Head trauma seems to be influenced by helmet use, with a nearly sixfold increase among non-users (RR = 5.73), while injuries to thorax, abdomen and extremity occurred with identical frequency in helmeted and unhelmeted scooter riders. After adjusting for possible confounders, helmet use has a protective effect in respect to head injury (OR = 0.23; 95% CI: 0.026-0.482). In the pre-post legem study we found that in 1999, injured not wearing a helmet represented 95% of the total injured, while in the two periods of 2000 this percentage dropped to 2.8% and 10% respectively. The number of accidents increased in the second period of 2000, but the number of lesions decreased, especially those affecting head (12.43% in 1999, 7.8% e 3.85% in the first and second period of 2000), and a sensible reduction in the use of head imaging diagnostics was also recorded. Mean length of stay for injured scooter riders was 7.1 days in 1999, 8.7 days in the first period of 2000, and 5.6 days in the second period of 2000. The surveillance studies confirmed that helmet use has a protective effect in all the periods considered reducing the probability of head trauma by 3-4 times. PMID- 12632992 TI - [Aids, physicians, Catholic Church]. AB - The latest AIDS' congress (Barcelona) reminded the world this dramatic situation. The shown data are remarkable: 5 million people of new infected in 2001, 68 million people could die in the next 20 years because of AIDS and the biggest part of them is living in the South of the world. There are two different kind of AIDS: the AIDS of rich people (2% of infected ones), who can reach the modern therapies that changed the course of the disease now curable out of hospital, and the AIDS of poor ones, without therapies and future. The political-economic effort of Western governments, of global fund anti-AIDS and of non governmental organizations now is not able to answer to this emergency in the right way. The lacking sensibility of Western doctors and the inflexible position of Catholic Church about contraception make the situation more complicated. It's hopeful the overcoming of this position using a Catholic Church's precious concept: the distinction between "simpliciter" and "secundum quid" to agree the use of condoms in case of absolute need. PMID- 12632993 TI - [An index for evaluation of cognitive-behavioral disorders in elderly people living in nursing homes]. AB - Aim of the study was to create an instrument (DISCO Index) to evaluate cognitive behavioural disturbances in elderly people. We considered n. 192 elderly people living in Torino's nursing homes, n. 76 people screened by the Geriatric Evaluation Unit (GEU) of S. Giovanni Battista Hospital in Torino and n. 136 subjects evaluated by a Torino's District GEU. Mean age was 81.8 +/- 8.3 years. By evaluating cognitive status and behavioural disturbances we obtained a list of conditions referring to three different degrees of severity depending on care needs. The three categories have been defined as: group A: absent or light cognitive and behavioural disturbances; group B: significant impairment of space timing orientation, hallucinations and delirium, frequent verbal abuse (outrage, menace), feeding alteration; group C: total disturbance of sleep--awakeness rhythm, frequent real or threatened physical assault, wandering or escape attempts, suicidal ideas or suicidal trials, sever feeding alterations. Functional status was also considered by ADL. 41.1% of the study population belongs to group A, 50.5% to group B and 8.4% to group C. Functional status in the sample was seriously impaired in 64.4% (autonomy loss in more then three functions) while only 9.4% of the subjects was autonomous. The totally of people belonging to the group C was found to be dependent in ADLs. PMID- 12632994 TI - [Infective endocarditis as predisposing factor to recurrence]. AB - In this paper we communicate our experience with 60 patients who suffered two or more episodes of infectious endocarditis (IE), selected from 1053 cases of IE defined in accordance with Duke's Hospital criteria. Relapsing IE was diagnosed when episodes occurred with at least a 6 month interval between them or when, even if the time distance was shorter, they were caused by different microorganisms. Sixty-four cases of relapsing IE were observed. In each patient we considered age, sex, presence of risk-factors for IE, time laps from the onset of symptomatic disease to diagnosis, time interval between the two or more IE episodes, aetiology, endocarditic location, echocardiographic elements, therapy, complications and final outcome were considered. We conclude, agreeing with other authors, that infectious endocarditis may rightfully be considered a risk factor for further IE episodes and that, for this reason, an adequate prophylactic therapy must be given. PMID- 12632995 TI - [Maculopathy in Curschmann-Steinert myotonic dystrophy]. AB - Besides hereditary macular dystrophies as well as acquired and age-related macular degenerations, systemic disorders should also be considered as a rare cause of maculopathy. In particular, myotonic dystrophy (Curschmann-Steinert syndrome), also characterized by the presence of ptosis and a pathognomic star shaped cataract, should be taken into account since a reticular dystrophy of the retinal pigment epithelium could be observed, resulting in a visual impairment of various degrees. A clinical case have been retrospectively reviewed and reported in this paper. PMID- 12632996 TI - [Cholestasis. Recent diagnostic and therapeutic acquisitions]. AB - New acquisitions in the field of physiopathology of biliary secretion and the improvement of methods for evaluating the biliary tree allowed a significant progress in the development of new diagnostic procedure and new therapeutical strategies for patients with cholestatic syndromes. The main novelties are here discussed. PMID- 12632997 TI - [Neutrophils in rheumatoid inflammation]. AB - For many years a secondary role in the pathogenesis of rheumatoid arthritis has been ascribed to neutrophil, relatively to the inflammatory response's evaluation. This cell was considered lacking in a peculiar activity and ever depending on lymphocytes and monocytes. During the recent years the neutrophil has been recognized as a cytokines producing cell, really able to modulate its role in joints inflammation. In the light of the latest information it's possible reconsider the role of this cell, looking at it like a moderate co-protagonist in the expression of rheumatoid damage, regarding both to joint inflammation and the maintenance of the damage itself. On the grounds of these knowledge, polymorphonuclear granulocyte could be also chosen as target of the newest therapies in the treatment of this disease. The aim of this short review is to focus the activity of neutrophils in rheumatoid arthritis, trying to follow them through their migration from blood to sinovial tissue and to understand the dynamic relation with the cytokine network, that from these cells pathway depends. PMID- 12632998 TI - [Current methods of glucose metabolism control in diabetes mellitus]. AB - The evaluation of glicometabolic control has been always considered important in the follow-up of the diabetic patient. Several methods are today available in order to register the short and long term glycaemic values. Recent studies have attributed to the post-prandial hyperglycaemias a specific role in determining the chronic complications. New pharmacological molecules are now available to reproduce the physiologic glycaemic profile. PMID- 12632999 TI - [What do medical students read?]. PMID- 12633000 TI - [A singular nonsense of Charles Ludwidge Dodgson (Lewis Carroll): mathematician, philosopher, philologer, scholar and refined humorist]. PMID- 12633001 TI - State substance abuse and mental health managed care evaluation program. AB - The articles in this special section of the Journal of Behavioral Health Services & Research (30:1) present results from evaluations of publicly funded managed care initiatives for substance abuse and mental health treatment in Arizona, Iowa, Maryland, and Nebraska. This overview outlines the four managed care programs and summarizes the results from the studies. The evaluations used administrative data and suggest a continuing challenge to structure plans so that undesired deleterious effects associated with adverse selection are minimized. Successful plans balanced risk with limited revenues so that they permitted greater access to less intensive services. Shifts from inpatient services to outpatient care were noted in most states. Future evaluations might conduct patient interviews to examine the effectiveness and quality of services for mental health and substance abuse problems more closely. PMID- 12633002 TI - The Iowa Managed Substance Abuse Care Plan: access, utilization, and expenditures for Medicaid recipients. AB - The Iowa Managed Substance Abuse Care Plan (IMSACP) used a behavioral health care organization to manage expenditures for treatment of alcohol and drug dependence financed through Medicaid, block grants, and state appropriations but maintained relatively distinct eligibility and benefit structures for Medicaid-eligible individuals. Medicaid claims, encounters, and eligibility files were reviewed for 2 years before and 3 year after implementation of IMSACP to evaluate changes in access, utilization, and expenditures. The rate of substance abuse treatment doubled, use of inpatient hospital services decreased, and residential and outpatient services increased. Direct care costs decreased, while total expenditures held steady. The Iowa experience suggests that a well-planned initiative can control costs and improve access and utilization. PMID- 12633003 TI - The impact of managed care on the substance abuse treatment patterns and outcomes of Medicaid beneficiaries: Maryland's HealthChoice program. AB - The introduction of Medicaid managed care raises concern that profit motives lead to the undersupply of substance abuse (SA) services. To test effects of the Maryland Medicaid HealthChoice program on SA treatment patterns and outcomes, Medicaid eligibility files were linked to treatment provider records and two study designs were used to estimate program impact: a quasi-experimental design with matched comparison groups and a natural experiment. Patient sociodemographic and clinical characteristics were adjusted using multiple regression. Under managed care, there was a shift from residential, correctional-only, and detoxification-only treatment toward outpatient-only treatment. Among beneficiaries entering treatment, those enrolled in managed care organizations (MCOs) had similar utilization and outcomes to those in Medicaid fee-for-service; those enrolling in MCOs during treatment had longer and more intensive episodes and, as a result, better outcomes. Thus, the study disclosed no empirical evidence that health plans respond to capitation by reducing SA services. PMID- 12633004 TI - Do adjusted clinical groups eliminate incentives for HMOs to avoid substance abusers? Evidence from the Maryland Medicaid HealthChoice program. AB - The adequacy of risk adjustment to eliminate incentives for managed care organizations (MCOs) to avoid enrolling costly patients had been questioned. This study explored systematic differences in expenditures between beneficiaries with and without substance disorders assigned to the same capitation rate group under the Maryland Medicaid HealthChoice program. The investigators used fiscal year (FY) 1995 to 1997 Medicaid data to assign beneficiaries to rate cells based on FY 1995 diagnoses and compared the distribution of expenditures for beneficiaries with and without substance disorders, defined using FY 1997 and FY 1995 diagnoses. Results showed that differences in FY 1997 expenditures between beneficiaries with and without FY 1995 substance disorders were negligible. However, MCOs could expect greater average losses and lower average profits on beneficiaries with FY 1997 substance disorders. Thus, the adjusted clinical groups methodology used to adjust capitation payments in the HealthChoice program attenuated, but did not eliminate, financial incentives for MCOs to avoid substance abusers. PMID- 12633006 TI - The Nebraska Medicaid managed behavioral health care initiative: impacts on utilization, expenditures, and quality of care for mental health. AB - This study evaluates the impact of Nebraska's Medicaid managed care program for behavioral health services on mental health service utilization, expenditures, and quality of care. Implementation of the program is correlated with progressive reductions in both total (about 13% over 3 years) and per eligible per month (20%) expenditures and a rapid, extensive decline in inpatient utilization and admissions. The percentage of enrollees receiving any type of treatment for a mental disorder actually increased modestly. Most important, several indicators of quality of care (e.g., timely receipt of ambulatory care following discharge from inpatient care and readmission to inpatient care shortly following discharge) suggest that quality of care did not materially change under the carve out. Although a thorough assessment of quality of care impacts is warranted, this study suggests implementation of a managed care program may allow states to reduce Medicaid expenditures without compromising quality of care. PMID- 12633005 TI - Evaluating selection out of health plans for Medicaid beneficiaries with substance abuse. AB - In the absence of adequate risk adjustment, capitation for enrollees creates incentives for health plans to enroll and retain good risks and to avoid bad risks. This article examines whether Maryland Medicaid beneficiaries with histories of substance abuse disenroll from health plans more frequently than those without such histories. The findings indicate that enrollees with a history of substance abuse were more likely to switch plans than other enrollees, regardless of whether they chose the health plan or were randomly assigned to the plan. These results suggest that current risk-adjustment systems may fail to offset selection incentives in modern capitated health plans. PMID- 12633008 TI - What proportion of failure to complete secondary school in the US population is attributable to adolescent psychiatric disorder? AB - Using information available from the longitudinal Children in Community Study, population-attributable risk percentage was calculated to estimate the amount of failure to complete secondary school in the United States that is associated with adolescent psychiatric disorder. Over half the adolescents in the United States who fail to complete their secondary education have a diagnosable psychiatric disorder. The proportion of failure to complete school that is attributable to psychiatric disorder is estimated to be 46%. School failure among young persons with psychiatric disorder exacts a large toll from individuals and society. PMID- 12633007 TI - The impact of evidence-based guideline dissemination for the assessment and treatment of major depression in a managed behavioral health care organization. AB - This study tests whether a managed behavioral health care organization can influence adherence to practice guidelines for the treatment of major depression in a randomized trial of guideline dissemination. Guidelines were disseminated to mental health clinicians (N = 443) under one of three conditions: (1) a general mailing of guidelines to clinicians, (2) a mailing in which guidelines were targeted to a patient starting treatment with the clinician, and (3) no mailing of guidelines. The results showed no effects of guideline dissemination as measured by self-report of patients and clinicians and through episode characteristics derived from claims data, despite sentinel effects. Results also showed high rates of clinician-reported guideline adherence that were not detected in the claims data, indicating significant undertreatment of depression. Results suggest that mental health systems must look to other dissemination strategies to improve adherence to standards of care and raise the performance of independent practicing clinicians. PMID- 12633010 TI - "The evil among us" or "the buck stops there". PMID- 12633009 TI - Case-mix adjustment of adolescent mental health treatment outcomes. AB - Case-mix adjustment methods are needed to account for differences between providers when the youth they treat have characteristics that adversely affect treatment success. This study explores variables for adjusting mental health treatment outcomes for adolescents and the differential effects of case-mix adjustment on providers' performance. Linear regression modeling was used to identify case-mix variables for five outcomes. Predictive equations for each outcome were developed for models based on intake clinical data alone, clinical data plus administrative data, and clinical data plus data describing youth history and family environment. Variance explained by intake clinical data alone did not increase appreciably with the addition of administrative data or data describing youth history and family environment. Adjusting outcomes changed the relative performance of certain individual providers substantially, but had a more moderate impact on the overall interpretation of providers' performance. PMID- 12633011 TI - The hidden vulnerable among us. PMID- 12633012 TI - Obstetricians & gynecologists--women's health care physicians. PMID- 12633013 TI - New contraception options. PMID- 12633014 TI - Chronic pelvic pain in women. PMID- 12633015 TI - Polycystic ovary syndrome and insulin resistance. PMID- 12633016 TI - Preconceptional counseling and care: a unique window of opportunity. PMID- 12633017 TI - Images in medicine. A torsed uterine fibroid presenting as a radiologic challenge. PMID- 12633018 TI - CMS National Surgical Infection Prevention Project: quality indicator rate for Rhode Island 2001. PMID- 12633019 TI - Infant mortality in Rhode Island: a time trend analysis. PMID- 12633020 TI - Reptile-associated salmonellosis: a preventable pediatric infection. PMID- 12633021 TI - Media: using 'bug chasers'. PMID- 12633022 TI - Getting the lead out. PMID- 12633023 TI - Is there a cure for cancer ads? PMID- 12633024 TI - The breaking point. PMID- 12633025 TI - Fight No. 2: docs vs. lawyers. PMID- 12633026 TI - Whistleblower. Nurses gain important protection. PMID- 12633027 TI - Covering the uninsured campaign. PMID- 12633028 TI - Violence towards the caregiver. A growing crisis for professional nursing. PMID- 12633029 TI - Medicaid reform on the table. An invitation we couldn't resist! PMID- 12633030 TI - Build the excitement when they're young. PMID- 12633031 TI - [Surgical treatment of hepatic hydatidosis. Analysis of results of conservative and radical surgery in our experience]. AB - The authors review a series of 194 cases of hepatic hydatidosis submitted to surgery over the period from July 1973 to December 2001, comparing the results of conservative and radical operatins. In this period we performed 164 conservative operations (140 indirect marsupialisations, 5 delaminations with capitonnage of the residual cavity and 19 partial pericystectomies), 26 total pericystectomies and 4 liver resections. The main outcome measures analysed were postoperative hospital stay, mortality, morbidity and recurrence rates. The median postoperative hospital stay was 17.5 days for conservative operations and 12.5 days for radical surgical procedures. No deaths occurred in either group. The morbidity was 26.8% after conservative treatment (34 biliary fistulas, 3 cases of jaundice, 5 residual cavity infections and 2 subphrenic abscesses) and 13.3% after total pericystectomy and liver resection (3 wound infections and 1 pleural effusion). Recurrences occurred only in the group treated by marsupialisation (12/140 = 8.5%) and were managed by total pericystectomy in 5 cases and by a second drainage procedure in 7. The surgical procedure for the treatment of hepatic hydatidosis should be tailored to the needs of each patient, depending on the size, location and complications of the cyst and high surgical risk should be avoided in view of the benign nature of the disease. No deaths occurred in either of our patient groups, but postoperative hospital stay, morbidity and recurrence rates were better in those undergoing radical treatment. PMID- 12633032 TI - [Pleuro-pneumonectomy, RT and CT for pleural mesothelioma. Prospective study]. AB - Though associated with substantially prolonged survival, the favourable results of Sugarbaker's trimodality treatment for malignant pleural mesothelioma are controversial and have yet to be extensively reproduced. The aim was to evaluate the prospective medium-term (3-year) reproducibility of the results of trimodality treatment in a significant group of patients staged using the international IMIG classification. Forty-three patients with malignant pleural mesothelioma were candidates for extended pleuropneumonectomy, followed by chemo- and radiotherapy. At thoracotomy, 33 of the 43 surgical candidates underwent extended pleuropneumonectomy and 71% of the 30 evaluable operated patients completed the scheduled course of adjuvant chemotherapy. The perioperative mortality rate was 6% and the major morbidity rate 36%. At 3 years the overall survival of the 30 evaluable patients was 30% and the disease-free survival rate was 25%, with a prevalence of epithelial pI and pII IMIG stages. Clinical understaging was appreciable. Survival was less favourable for stage pIII and for mixed tumours. Most of the disease recurrences were due to incomplete local postsurgical monitoring. This series confirms the reproducibility of trimodality treatment for malignant pleural mesothelioma. The treatment is associated with prolonged survival in the case of early-stage tumours and has an acceptable complication rate. Early diagnosis, accurate staging, preoperative induction and better local monitoring are avenues to be explored when seeking to achieve curability of malignant pleural mesothelioma. PMID- 12633033 TI - [Current trends of artificial enteral nutrition in acute pancreatitis]. AB - During recent years, there has been considerable debate as to the nutritional supply that needs to be established for a patient with acute pancreatitis. The main problem is still infection of the pancreatic necrosis, which has a decisive bearing on the indication for surgery and is the main cause of mortality. Infection stems from bacterial translocation from the patient's gut. Enteral nutrition with its known potential for reducing this type of infection constitutes an attempt to prevent it by preserving the enteric mucosal barrier. Today, the concept of pancreatic rest is no longer considered mandatory in the guidelines of many Surgical and Nutritional Societies, whilst enteral nutrition is the gold standard for acute pancreatitis. Assuring an integrated parenteral and enteral supply before reaching the full regimen of enteral nutrition is the most reliable policy during the early days of the disease. Moreover, outcomes being equal, enteral nutrition is cheaper than parenteral nutrition, as has been extensively demonstrated in many clinical trials in severe acute pancreatitis. PMID- 12633034 TI - Adrenal incidentalomas: indications for surgery. AB - Adrenalectomy represents the gold standard treatment for hyperfunctioning adrenal incidentaloma. In cases of silent adrenal masses, on the other hand, the surgical removal of an adrenocortical cancer entails the sacrifice of a large number of safe benign masses, and in most cases surgery is therefore unjustified. The aim of this paper was to clarify the surgical indications for adrenal incidentaloma by reviewing our experience in comparison with the main reference literature. Over the period from 1995 to 2001 we managed 40 cases of incidentaloma. US and CT abdominal scans, adrenal scintigraphy and biochemical tests were performed on an outpatient basis. Seven pre-Cushing syndromes were removed. Ten incidentalomas measured 4 cm or more in diameter: 5 of these were operated on and in 5 cases surgery was not feasible or was refused. Only one malignant mass was detected (an angiosarcoma). Four postoperative minimal complications (18.7%) were observed. The follow-up (median: 48 months) was uneventful. The surgical approach was traditional in 11 cases and laparoscopic in 1 case. Surgery should be considered mandatory in cases of hyperfunctioning adrenal masses in the presence of suspect radiological evidence, in cases of discordant CT and scintigraphy findings and when the maximum diameter is 4 cm or more. PMID- 12633035 TI - Factors predicting outcome of hypocalcaemia following total thyroidectomy. AB - Postoperative hypocalcaemia is often observed after total thyroidectomy. In patients requiring calcium replacement therapy after 1 year, hypocalcaemia must be considered permanent. The aim of this study was to assess the incidence of hypocalcaemia following total thyroidectomy and to evaluate the risk factors predicting delayed outcome such as hypoparathyroidism. From January 1998 to September 2001, 310 patients underwent total thyroidectomy in our department. In a total of 37 patients experiencing hypocalcaemia, the authors carried out a comparative study of 34 patients with transient hypocalcaemia (group A) and 3 patients with permanent hypocalcaemia (group B). The incidences of transient and permanent hypocalcaemia were 11.9% and 0.9%, respectively. Central neck lymph node dissection performed in cases of thyroid carcinoma correlated with permanent hypoparathyroidism. The most significant factors predicting long-term outcome of hypocalcaemia were low serum calcium levels (< 8 mg/dl) and high serum phosphorus levels (> 5 mg/dl) measured on postoperative day 7, despite oral calcium replacement. The indications for lymph-node dissection in the central neck area should be very strictly selected. When delayed serum calcium and phosphorus levels are unfavourable, thorough follow-up of patients is mandatory in order to administer the correct therapy and prevent the consequences of chronic hypocalcaemia. PMID- 12633036 TI - [Bile duct injuries due to unwarranted duct catheterization during cholangiography (prevention and treatment)]. AB - Attention is drawn to bile duct injuries resulting from unwarranted duct catheterism during cholangiography both for a presumed cystic duct, which turns out not to be one, and for a true cystic duct downstream of an aberrant duct which unfortunately is only detected later. The importance is stressed of access to the cystic duct after cautious dissection, extended to the gallbladder, has detected it up to the vent and ruled out the existence of an aberrant duct. There are several potential options available for the treatment of such ductal lesions, analysing their impact on the parenchyma. The surgeon should not entrust the patient's safety solely to the contribution of cholangiography. Recognition of mistreatment may come too late for effective remedial measures to be taken, particularly in those injuries which are not amenable to conservative treatment. PMID- 12633037 TI - [Acute vascular abdomen: personal experience and review of the literature]. AB - Acute vascular abdomen presents an incidence of 1% of all cases of acute abdomen, but the mortality rate is high, ranging from 50 to 80%. Acute mesenteric ischaemia and rupture of abdominal aorta aneurysms are the most frequent causes. Prevention of causal agents and early diagnosis are the only elements capable of improving patient survival. The authors report their personal experience over the period from 1996 to June 2002, analysing different clinical cases of acute vascular abdomen and their diagnostic and therapeutic management and comparing their experience with the relevant literature data. PMID- 12633038 TI - Periumbilical veress needle pneumoperitoneum: technique and results in 2.126 cases. AB - In laparoscopic surgery, a pneumoperitoneum is generally created either by direct puncture (Veress needle technique) or by the open (Hasson) technique. Some surgeons predominantly use direct access, reserving open access for selected patients; others consider the Veress needle as carrying an unacceptable risk of intestinal or vascular injury and consequently use only the Hasson technique. The database containing records of all abdominal laparoscopic operations performed in our institution was reviewed; over the period from June 1991 to December 2001 a total of 2,126 laparoscopic operations were performed, consisting in 1,457 (68.5%) cholecystectomies, 392 (18.4%) appendectomies, 177 (8.3%) varicocelectomies, 65 (3%) operations on the uterus and ovaries, 6 (0.2%) groin hernia repairs, 6 (0.2%) fundoplications, 3 (0.1%) ileo-colic resections for right colon cancer, and 20 (1.3%) diagnostic laparoscopies. The direct puncture technique was used in 100% of cases. The time needed up to connection of the video camera averaged 4-7 minutes. Twelve laparoscopic cholecystectomies were converted, with a conversion rate of 0.5%. Four patients were re-operated on, two for bleeding from the gallbladder fossa and two because of small bowel perforation caused by electrocautery. None of the patients sustained any clinically apparent vascular or bowel injury as a result of the blind access. There were no cases of preperitoneal or omental air insufflation. The mortality was nil. The authors describe their technique for gaining access to the abdominal cavity by direct puncture and conclude that properly performed Veress needle insertion is less invasive, safe and perfectly feasible in all patients. PMID- 12633039 TI - Laparoscopic management of non-parasitic splenic cysts. AB - Non-parasitic cysts are an infrequent disease of the spleen. Most likely congenital in their origin, epithelial splenic cysts require surgical treatment if enlarged or symptomatic. The aim of this study was to assess the impact of the laparoscopic technique in the management of splenomegaly due to cystic disease. From July 2001 to September 2002 4 patients underwent laparoscopic splenectomy in our department for symptomatic enlarged non-parasitic splenic cysts. Cystic lesions were evaluated by ultrasonography, CT scan and magnetic resonance of the abdomen. The choice of total laparoscopic splenectomy according to the "hanged spleen" procedure was conditioned by the technical difficulty of performing conservative treatment due to the site of the upper pole of the enlarged lesions, adherent to the diaphragm in 3 cases, and by the presence of multiple trabeculated splenic cysts in 1 case. Laparoscopy was always completed and all patients obtained relief of their symptoms. When conservative treatment such as partial splenectomy or decapsulation may jeopardize the good outcome of the operation, the management of nonparasitic splenic cysts can be successfully achieved by total laparoscopic splenectomy. PMID- 12633041 TI - [Organization of a day surgery unit for hernia]. AB - The authors report their experience with the organisational aspects of minimal hospitalisation of patients undergoing surgical treatment for hernias of the abdominal wall. 1544 hernia repairs were carried out from January 1994 to March 2002; over 80% of these were performed on a day surgery basis. The organisation of a day surgery unit requires dedicated medical and nursing staff and also diagnostic imaging facilities that should be immediately available, though all this is not strictly necessary in a general surgery department. In conclusion, in addition to the organisational problems, inadequacy of the D.R.G. system in Italy is the primary obstacle to diffusion of the practice of ambulatory hernia repair. PMID- 12633040 TI - [Acute cholecystitis: emergency treatment, videolaparoscopic technique and indications for conversion]. AB - The authors discuss the present approach to acute cholecystitis and the difficulties encountered in videolaparoscopic treatment. The effective usefulness of this new approach needs to be very carefully evaluated and the surgeon must bear in mind the patient's safety and the surgical risk. Therefore, in cases of very serious difficulties or increased risk, it is mandatory to convert the procedure to an open cholecystectomy. PMID- 12633042 TI - Pelvic actinomycosis masquerading as malignancy: report of two cases and review of the literature. AB - Pelvic actinomycosis is uncommon, presenting most frequently as an abdominal mass and simulating advanced malignancy in female patients with a past history of intrauterine contraceptive use. It responds favourably to prolonged antibiotic therapy with occasional need for abscess drainage and debridement or ureteric stenting. Incorrect diagnosis may result in overly invasive investigations and unnecessarily radical extirpative surgery. PMID- 12633043 TI - [Superior mesenteric venous thrombosis. Report of 2 cases and review of the literature]. AB - Mesenteric venous thrombosis is an uncommon but often fatal form of intestinal ischaemia. The authors present two cases of severe small intestinal ischaemia due to superior mesenteric vein thrombosis, the first associated with oral contraceptive use and a congenital methylene-tetrahydrofolate reductase defect and the second associated with atypical intestinal mycobacterial disease and acquired immunodeficiency syndrome. The authors review the relevant literature using the Medline search facility and comment on the changing aspects of MVT syndrome with regard to aetiological factors, diagnostic approach and surgical or pharmacological therapy. PMID- 12633044 TI - [Inflammatory aneurysm of the abdominal aorta and its complications: report of 2 clinical cases]. AB - The authors present two cases of inflammatory abdominal aortic aneurysms complicated by an aorto-caval fistula in one case and by an aorto-duodenal fistula in the other. The aetiology of such aneurysms is still debatable, the histological features are typical, and the preoperative clinical and instrumental findings do not always allow them to be differentiated from atherosclerotic ones. The complications may be rapidly fatal or asymptomatic, as in the case of occult aorto-caval fistulas. The surgical approach is technically more difficult because of inflammation and adhesions between the aneurysmal sac and contiguous structures which make dissection and aorto-iliac clamping more demanding. In the case of aorto-caval fistulas, clamping of the vein should be avoided and manual compression preferred to assure haemostasis; with aorto-enteric fistulas, it is necessary to reduce the contamination of the operative field. Ureterolysis is not always necessary in patients with hydronephrosis. Endovascular devices may be an alternative also for a number of complications. PMID- 12633045 TI - [Etiopathogenic and clinico-therapeutic considerations on Spigelian lateral ventral hernia. A caseload and review of the literature]. AB - The authors report on 6 cases of Spigelian hernia surgically managed over the period from 1973 to 2002. They describe the anatomy of the anterolateral abdominal wall and the etiopathogenetic factors of this rare type of hernia. The clinical features of the disease are discussed and the diagnostic usefulness of a thorough physical examination is stressed, as is the value of imaging procedures when the hernia is not evident. They, then, go on to describe the repair techniques currently available that are capable of yielding an excellent long term outcome. PMID- 12633046 TI - Well-differentiated giant "lipoma-like" liposarcoma of the posterior mediastinum: a case report. AB - While liposarcoma is the most common malignant mesenchymal neoplasm in adults, a mediastinal position is rare. We describe here the case of a 74-year-old female with an hourglass-shaped mass in the posterior mediastinum, which was probably present on a roentgenogram 4 years earlier. On a CT scan of the chest, the mass appeared non-homogeneous with a mainly adipose content. A preoperative ultrasonically guided tru-cut transthoracic biopsy revealed the presence of adipose tissue with mildly atypical cytological features. The mass was therefore completely excised by means of a right thoracotomy. The postoperative course was uneventful and the patient is alive and well and has had no recurrence in the 6 months since surgery. We review here the natural history, pathology, and prognosis of the disease, and discuss methods of diagnosis and management of such lesions. PMID- 12633047 TI - Mucinous cystic lesions of the pancreas. Report of a patient submitted to distal pancreatectomy. AB - Intraductal papillary neoplasm consists a variant of the mucin-producing neoplasms of the pancreas; these are rare diseases that can be divided into mucinous ductal ectasias and intraductal papillary neoplasms. Such neoplasms have a malignant potential, and their prognosis is good if they are early diagnosed and localized surgical resection. This paper describes a case of an intraductal papillary neoplasm localized in the tail of the pancreas and treated by distal splenopancreatectomy. The literature on this rare disease is also reviewed. PMID- 12633048 TI - [Pancreatic neck rupture: report of a clinical case]. AB - The authors report a case of pancreatic neck rupture in a 45-year-old female patient due to blunt trauma in a car crash. An emergency laparotomy was performed and the pancreatic head side was sutured, whereas the left side was anastomosed to a Roux-en-Y jejunal loop. The aim of this report is to emphasize the advantages of conservative surgery with internal drainage. This technique has been described for benign pancreatic tumours and protects the patient against exocrine and/or endocrine insufficiency. PMID- 12633049 TI - [Treatment of esophageal perforations. Considerations on a clinical case]. AB - The Authors describe a rare case of esophageal perforation occurred after Transoesophageal echocardiography in 68 years old patient and review the literature relating to the causes and management of this pathology. Transoesophageal echocardiography, which is a semi-invasive investigation increasingly used in cardiology and cardiac surgery and intensive care units, is a rare though extremely dangerous cause of such complications. Perforation of the esophagus continues to present a formidable diagnostic and therapeutic challenge. The diagnosis depends on a high degree of suspicion and on the recognition of clinical features and is confirmed by contrast esophagography. The outcome after esophageal perforation depends on the location of the injury, the presence or otherwise of concomitant esophageal disease and the time elapsing between the injury and inititian of treatment. Reinforced primary repair of the perforation is the procedure most frequently employed and preferred for the surgical management of the esophageal perforation. In the case reported here, early diagnosis and prompt surgical treatment consisting in primary repair of the esophageal perforation contributed to the successful management of this serious pathology. PMID- 12633050 TI - [Complete surgical resection of a huge thoracic mass consisting in a localized fibrous tumor simulating a high-grade malignant neoplasm]. AB - Solitary fibrous tumour is an infrequent neoplasm generally arising from the parietal and visceral pleura. The diagnosis may be difficult in the presence of a history of malignant disease owing to the different presentations and to radiological findings of evident invasiveness. The authors report the case of a woman with a right giant fibrous solitary tumour of the pleura twenty years after a subcutaneous mastectomy with axillary dissection and radiation therapy for breast cancer. The biopsy diagnosis was consistent with a probable solitary fibrous tumour of the pleura but the discrepancy with the radiological images and the difficult differential diagnosis versus a malignant sarcoma, possibly radio induced, prompted us to verify the real features of the disease. The patient was submitted to a right anterolateral thoracotomy and partial sternotomy and the giant mass was resected enbloc with the phrenic nerve and diaphragm which proved to be the only structures tightly adhering to the neoplasm. Histological examination confirmed the diagnosis of a solitary fibrous tumour of the pleura. The patient is still alive and disease-free 30 months after the surgical operation. Fibrous solitary tumour is a disease generally characterised by a good prognosis but in particular cases, with unmistakable radiological findings of invasiveness, a precise diagnosis must be obtained in order to choose the most appropriate therapy. PMID- 12633051 TI - [A case of tubercular peritonitis: diagnostic value of laparoscopy]. AB - A case of tubercular peritonitis is illustrated in which the diagnosis is made thanks to the use of diagnostic imaging. Diagnostic certainty, however, can be obtained only by laparoscopy, which provides both a direct view of the lesions and biopsy samples of the peritoneal lesions. PMID- 12633052 TI - The treatment of large midline incisional hernias using a retromuscular prosthetic mesh (Stoppa-Rives technique). AB - Incisional hernia is a common problem after abdominal surgery. Many repair techniques with prosthetic meshes have been proposed but there is no general agreement as to the best choice. Our retrospective experience with 35 patients treated using a large polypropylene mesh placed beneath the rectus muscles and above the peritoneum (Stoppa-Rives technique) is reported. There was no operative mortality. Major postoperative complications occurred in 7 (20%) patients. Wound infection developed in 5 (14.2%) patients and in one case the mesh had to be removed. The recurrence rate was 2.8%. In conclusion, retrorectus preperitoneal mesh repair is an effective technique with a low recurrence rate and very few postoperative complications. PMID- 12633053 TI - [The use of autologous dermis in the treatment of incisional hernia]. AB - Incisional hernia is a relatively frequent complication of laparotomy. In reconstruction of large abdominal wall defects prosthetic meshes have come to play a leading role. However, they carry a high risk of infection and are poorly effective in restoring perfect chest-wall function. In order to achieve a better result we treated two cases of incisional hernia with a combined technique, namely, abdominoplasty with subcutaneous implantation of in-vitro cultured autologous dermis (Hialograft 3D). As described in the literature, we noticed that this kind of implant integrates perfectly with the receiving tissues with no sign of rejection or reabsorbtion. It provides good support for the abdominal wall with excellent elastic properties that make for better restoration of chest wall function compared to prosthetic implants. Two years after the operation there are no signs of recurrence. The high cost and the time needed for culturing the fibroblasts are the main drawbacks of this technique. Despite the very small sample size and short follow-up, the encouraging results obtained to date justify further study. PMID- 12633054 TI - [Repair of mucosa stomal prolapse with circular stapler. Technical note]. AB - Mucosal stomal prolapse is a frequent complication of end colostomies. We describe here an original mucosal colostomy prolapse repair technique using a circular stapling device. This technique stems from the recent introduction of stapled muco-haemorrhoidectomy and was employed on a 68-year-old male patient who had undergone a Miles abdominoperineal excision 17 years earlier. On clinical inspection the patient presented a 5-cm. mucosal stomal prolapse on straining. The surgical procedure was performed under i.v. sedation with midazolam and propofol. After draping the abdomen, the stomal mucosa was gently everted with the aid of three Allis forceps, and a 1.0 prolene purse string suture was created 4-6 cm. cephalad to the cutaneous rim of the stoma. The head of a 33-mm circular stapler was introduced and the purse string suture was tied. The mechanical stapler was prepared and fired, thus obtaining a resection anastomosis of the prolapsed stomal mucosa. No bleeding occurred. A 24-F bladder catheter was placed cephalad to the stapled line and removed on postoperative day one. The patient was discharged from hospital the day after surgery. During a 16-month follow-up neither complications nor relapses have so far been observed. Our technique appears to be simple and safe. We believe it is indicated only for mucosal stomal prolapses, while full-thickness stomal prolapses should be managed by conventional techniques. A larger cohort of patients is needed to confirm our preliminary results. PMID- 12633055 TI - Testicular fixation in adult retractile testis: technical notes. AB - A variety of techniques for scrotal orchiopexy have been described in the literature, including those without suture fixation, such as the scrotal pouch. We perform a simple modified technique for bilateral testicular fixation in patients with bilateral and/or unilateral adult retractile testis and with symptoms characterised by chronic orchialgia due to testicular hypermobility or repeated funicular subtorsion. PMID- 12633057 TI - Partial universal coverage. PMID- 12633058 TI - Creating political will. PMID- 12633056 TI - Prospective randomized trial comparing implantation of subcutaneous indwelling venous access devices by the cephalic vein cutdown (CVC) approach versus the percutaneous subclavian vein (PSV) approach. PMID- 12633059 TI - Quality. A national effort for quality reporting. PMID- 12633061 TI - Compliance. HIPAA hangover. PMID- 12633060 TI - Quality. Facing the music. PMID- 12633062 TI - Security. Protect thyself. PMID- 12633064 TI - Disease management. The next step. PMID- 12633063 TI - Mission. Serving the community--near and far. PMID- 12633065 TI - 1Q[3a]. What are the hurdles in caregiver smallpox inoculations? PMID- 12633066 TI - Neal's zeal. PMID- 12633067 TI - MDs: [missing doctors?]. AB - Hospitals in places as varied as West Virginia, Missouri, Arizona and southern California say recruiting and keeping physicians are big problems, and getting bigger. But the evidence and the experts differ over whether a nationwide physician shortage on the scale of the nursing shortage is emerging or whether it is limited to specific markets. PMID- 12633068 TI - Health care benchmarking 2003. AB - The latest benchmarking data reveal both old trends and some surprises. Once again, hospitals in high managed care markets outperformed those in low managed care markets in profitability measures, such as total margin and return on equity. However, hospitals in areas with low managed care penetration had a lower percentage of bad-debt expense and significantly fewer days in accounts receivable and more days cash on hand. PMID- 12633069 TI - Putting patient safety in the blueprint. AB - When St. Joseph's Community Hospital of West Bend (Wis.) decided to build a replacement facility, CEO John Reiling proposed a novel approach: Let improving patient safety guide the entire design process. Before ground was broken, St. Joseph's convened a learning lab that attracted some of the nation's leading experts on patient safety. Since then, staff from every sector of the hospital have helped to identify where errors occur, how work flow and processes can change to enhance safety and how a building can be designed to incorporate those improvements. Although St. Joseph's leaders say the goal is simply to provide the best facility for their community, it's likely that their patient-safety-driven design process will become a model for hospitals across the country. PMID- 12633070 TI - Turning the corner. PMID- 12633071 TI - Synthesis of the mitomycin and FR900482 ring systems via dimethyldioxirane oxidation. AB - [reaction: see text] Dimethyldioxirane oxidizes a 2,3-dihydo-1H-pyrrolo[1,2 a]indole unsubstituted at the C-9 position stereoselectively to form a hydroxy ketone with all the basic elements of the mitomycin ring system. On the other hand, a 2,3-dihydo-1H-pyrrolo[1,2-a]indole derivative substituted with an alkyl group at C-9 undergoes an oxidative ring expansion in the presence of dimethyldioxirane to give an FR900482 analogue. PMID- 12633072 TI - Relative rate profile for ring-closing metathesis of a series of 1-substituted 1,7-octadienes as promoted by a 4,5-dihydroimidazol-2-ylidene-coordinated ruthenium catalyst. AB - [reaction: see text] This report details the kinetic responses of nine compounds of type 6 to ring-closing metathesis as promoted by 2 to give the identical product 7. The experimental observations have been subjected to Hammett analysis. The rho value for the composite aromatic derivatives (R = p-XC(6)H(4)-) differs from that of the aliphatic series, although both are negative because electron donating groups accelerate the reaction. PMID- 12633073 TI - Mild and efficient copper-catalyzed amination of aryl bromides with primary alkylamines. AB - [reaction: see text] An efficient copper-catalyzed amination of aryl bromides with primary alkylamines was developed that uses commercially available diethylsalicylamide as the ligand. This amination reaction can be performed at 90 degrees C in good yield. A variety of functional groups are compatible with these reaction conditions. Preliminary results show that this reaction can be carried out under solvent-free conditions with comparable yields. PMID- 12633074 TI - Synthesis of dihydrofurocoumarins via palladium-catalyzed annulation of 1,3 dienes by o-iodoacetoxycoumarins. AB - [reaction: see text] A variety of biologically interesting dihydrofurocoumarins have been synthesized in high yields by the palladium-catalyzed annulation of 1,3 dienes by o-iodoacetoxycoumarins. This reaction is very general and regioselective, and a wide variety of terminal, cyclic, and internal 1,3-dienes can be utilized. PMID- 12633075 TI - Heteroaromatic thioether-organostannane cross-coupling. AB - [reaction: see text] Heteroaromatic thioethers and aryl, heteroaryl, and alkenylstannanes participate in a palladium-catalyzed, copper(I)-mediated cross coupling reaction at 50 degrees C in THF. PMID- 12633076 TI - Copper(I)-promoted palladium-catalyzed cross-coupling of unsaturated tri-n butylstannane with heteroaromatic thioether. AB - [reaction: see text] Palladium-catalyzed cross-coupling of vinyl- and arylstannanes with pi-electron-deficient heteroaromatics was performed in good yields. This Stille-type reaction was carried out with a methylthioether function as an electrophile in the presence of a copper(I) bromide-dimethyl sulfide complex. PMID- 12633078 TI - 1,2-cyclic sulfamidates as versatile precursors to thiomorpholines and piperazines. AB - [reaction: see text] 1,2-Cyclic sulfamidates undergo regiospecific nucleophilic displacement with either methyl thioglycolate or alpha-amino esters, followed by lactamization (thermal, base-mediated, or cyanide-catalyzed), to give thiomorpholin-3-ones and piperazin-2-ones. PMID- 12633077 TI - Synthesis of 2'-C-beta-fluoromethyluridine. AB - [reaction: see text] 2'-C-beta-Fluoromethyluridine (17) represents both a potentially important biological agent and a tool for biochemical analysis. Here we describe the first synthesis of this compound starting from uridine. The key steps include protection of the uracil base with methoxyethoxymethyl (MEM) chloride, conversion to the corresponding 2'-C-alpha-epoxide, and regioselective opening of the oxirane ring with potassium fluoride/hydrogen fluoride. Subsequent acetylation of the 3'- and 5'-hydroxyl groups enables MEM removal using B bromocatecholborane. Deacetylation generates the parent nucleoside, 2'-C-beta flurormethyluridine. PMID- 12633079 TI - P[N(i-Bu)CH2CH2]3N: a versatile ligand for the Pd-catalyzed amination of aryl chlorides. AB - [reaction: see text] Palladium-catalyzed amination reactions of aryl chlorides with amines proceeded in the presence of the bicyclic triaminophosphine P[N(i Bu)CH(2)CH(2)](3)N to afford the corresponding arylamines in good to excellent yields. Electron-poor, electron-neutral, and electron-rich aryl chlorides all participated with equal ease. PMID- 12633080 TI - Chiral-auxiliary-controlled diastereoselectivity in the epoxidation of enecarbamates with DMD and mCPBA. AB - [structure: see text] Chiral oxazolidinone-substituted enecarbamates 1 are epoxidized in a diastereoselectivity up to 93:7 for both DMD and mCPBA. The diastereofacial differentiation depends on the steric interaction between the R(1) substituent on the oxazolidinone ring and the incoming electrophile. The stereochemical course of epoxidation was assessed by chemical correlation with the known optically active diols. PMID- 12633081 TI - Synthesis and studies of 1,4,5,8,9,12,13,16-octamethoxytetraphenylene. AB - [structure: see text] The synthesis of 1,4,5,8,9,12,13,16 octamethoxytetraphenylene (6) is accomplished in five steps from 3,6-dimethoxy-2 nitroaniline (8). Its inclusion property and the electrochemical data of the corresponding tetraquinone 7 are presented. PMID- 12633082 TI - Efficient production of enol ether radical cations by heterolytic cleavage of beta-mesylate radicals. AB - [reaction: see text] alpha-Methoxy-beta-mesyloxy radicals were produced in laser flash photolysis reactions, and yields of enol ether radical cations formed by heterolytic fragmentation of the mesylate group were determined. The mesylate heterolysis reaction is faster than heterolyses of phosphate and bromide groups in analogous radicals and highly efficient in medium-polarity solvents. PMID- 12633083 TI - Dearomatizing cyclization of arylsulfonylalkoxymethyl lithiums: a route to the podophyllotoxin skeleton. AB - [reaction: see text] The phenylsulfonyl group promotes the dearomatizing cyclization of tethered organolithiums onto aromatic rings. With an ether tether, the cyclizations create a new tetrahydrofuran ring, and both cyclization and subsequent electrophilic quenches proceed with high levels of diastereoselectivity. The sulfonyl group can be removed from the cyclized products oxidatively or reductively. The dearomatizing cyclization of a naphthyl sulfone was used in the synthesis of a close structural analogue of podophyllotoxin. PMID- 12633084 TI - Palladium-catalyzed enantioselective oxidation of alcohols: a dramatic rate acceleration by Cs2CO3/t-BuOH. AB - [reaction: see text] The addition of Cs(2)CO(3) and t-BuOH provides a dramatic rate acceleration in the palladium-catalyzed aerobic oxidative kinetic resolution of secondary alcohols while maintaining the selectivity of the process. PMID- 12633085 TI - Diphenylamino group as an effective handle to conjugated donor-acceptor polymers through electropolymerization. AB - [reaction: see text] The diphenylamino group is an effective handle for electropolymerization to give electron donor-acceptor conjugated polymers. In addition, interesting electrochromic and photoresponsive behavior of 13 has been investigated. PMID- 12633086 TI - Synthesis of the benz[a]anthraquinone core of angucyclinone antibiotics. AB - [reaction: see text] A general method for the synthesis of benz[a]anthraquinones is reported. The key step is a catalytic cobalt-mediated [2+2+2]-cycloaddition of a triyne, which affords an angularly substituted tetracycle. Oxidation of this core gives the typical structure of angucyclinone antibiotics. PMID- 12633087 TI - Cyclocarbopalladation: formation of bicyclic 1,2-cyclobutanediols through a rare 4-exo-dig cyclization. AB - [reaction: see text] Several bicyclic compounds bearing a strained 1,2 cyclobutanediol have been prepared from a gamma-bromopropargylic diol under palladium(0) catalysis. The reaction proceeds through a rare unfavored 4-exo-dig cyclocarbopalladation. In some cases, the first reaction is followed by a 6pi electrocyclization leading to unusual strained tricyclic systems. PMID- 12633088 TI - Solid-supported cyclohexane-1,3-dione (CHD): a "capture and release" reagent for the synthesis of amides and novel scavenger resin. AB - [reaction: see text] A three-step synthesis of cyclohexane-1,3-dione (CHD) resin 6 on polystyrene resin is described. Resin 6 was used to prepare an amide library of high purity by microwave-assisted serial "capture and release" and can be recycled for this purpose. High-loading CHD resin 10 was also shown to scavenge allyl cations in solution. PMID- 12633090 TI - Total synthesis of (+/-)-trachyspic acid and determination of the relative configuration. AB - [reaction: see text] The first total synthesis of (+/-)-trachyspic acid, a tumor cell heparanase inhibitor, was accomplished based on Cr(II)/Ni(II)-mediated reaction of the aldehyde containing the citric acid moiety and the long-chain triflate, and the relative configuration of this natural product was determined. PMID- 12633089 TI - Polymer-supported o-benzyl and o-allylisoureas: convenient preparation and use in ester synthesis from carboxylic acids. AB - [reaction: see text] Polymer-supported O-methyl, O-benzyl, and O-allyl-isoureas were prepared by copper(II)-catalyzed reaction of polymer-supported carbodiimide with the corresponding alcohols. These polymer-supported reagents were successfully employed to convert a series of carboxylic acids to methyl, benzyl, or allyl esters, in good yields. The products were obtained with high purity (>95% by NMR) after a simple resin filtration-solvent evaporation sequence. PMID- 12633091 TI - Transition metal-catalyzed regio- and stereoselective aminobromination of olefins with TsNH2 and NBS as nitrogen and bromine sources. AB - [reaction: see text] A new synthetic procedure for aminohalogenation of olefins has been developed for the preparation of vicinal haloamine derivatives in high yields by using Cu, Mn, or V catalysts with p-toluenesulfonamide (TsNH(2)) and N bromosuccinimide (NBS) as nitrogen and bromine sources, respectively. Unprecedented regio- and stereoselectivity (anti:syn > 99:1) toward the aminohalogenation process is shown for olefinic substrates as well as transition metal catalysts. PMID- 12633092 TI - How aromatic are large (4n + 2)pi annulenes? AB - [structure: see text] While the total aromatic stabilization energies (ASE) of the [n]annulenes, from C(6)H(6) to C(66)H(66), converge to ca. 22 kcal/mol, the ASEs per pi-electron decrease markedly. Bond length alternation (which depends on the theoretical level) only reduces stabilization somewhat but influences the magnetic properties (NICS, proton chemical shifts, and magnetic susceptibilities) considerably. Nevertheless, these magnetic criteria, when based on the most realistic structures, agree that the aromaticities of the larger annulenes decrease and then nearly vanish. PMID- 12633093 TI - Conversion of the enantiomers of spiro[4.4]nonane-1,6-diol into both epimeric carbaspironucleosides having natural C1' absolute configuration. AB - [reaction: see text] The enantiomers of spiro[4.4]nonane-1,6-diol have been transformed by different reaction pathways into the two possible carbaspironucleoside epimers with natural C1' absolute stereochemistry. PMID- 12633094 TI - Carbohydrate-derived spiroketals: stereoselective synthesis of di-D-fructose dianhydrides via intramolecular aglycon delivery. AB - [reaction: see text] A one-pot synthesis of C(2)-symmetric di-d-fructose dianhydrides having the 1,6,9,13-tetraoxadispiro [4.2.4.2]tetradecane skeleton has been accomplished via intramolecular aglycon delivery from (6 --> 6) xylylene tethered fructofuranose precursors. The stereochemical outcome of the glycosylation-spiroketalization process is governed by the geometrical constraints imposed by the rigid tetracyclic structure of the final compound. PMID- 12633095 TI - Naphthalenes, isoquinolines, and a benzazocine from zirconocene-copper-mediated coupling of benzocyclobutadiene with nitriles and alkynes. AB - [reaction: see text] Commercially available 1-bromobenzocyclobutene is a potentially useful synthon particularly with the application of organometallic methodology. Here we show that it is readily converted into Cp(2)Zr(benzocyclobutadiene), which couples with alkynes or nitriles giving five membered zirconacycles. Treatment of these alkyne- or nitrile-derived zirconacycles with CuCl yields substituted naphthalenes, isoquinolines, or in the presence of MeO(2)C-CC-CO(2)Me, a 3-benzazocine containing an eight-membered ring [corrected]. PMID- 12633096 TI - Highly regio- and chemoselective palladium-catalyzed three-component assembly of arylethylidene malononitriles, allylic chlorides, and allenylstannanes: a novel route to 1,7-enyne derivatives. AB - [reaction: see text] An efficient method for the synthesis of 1,7-enyne derivatives via a palladium-catalyzed three-component assembly of arylethylidene malononitriles, allylic chlorides, and allenylstannanes is described. PMID- 12633098 TI - Establishment of steric substituent constants in the adamantane system by ab initio calculations. AB - [reaction: see text] Steric substituent constants, S(A), of both alkyl and aryl substituents were calculated in the adamantane system by both isodesmic reactions and ab initio calculations. The method provides an easy and reliable way to quantify the substituent steric effects. PMID- 12633097 TI - Production of octadienal in the marine diatom Skeletonema costatum. AB - [reaction: see text] Marine diatoms produce alpha,beta,gamma,delta-unsaturated aldehydes that have detrimental effects on the reproduction of their natural predators. The production of these defensive metabolites is suggested to involve enzymatic oxidation of polyunsaturated fatty acids. In this paper, feeding experiments with labeled precursor provide clear evidence in support of the origin of octadienals 1 and 2 from 6,9,12-hexadecatrienoic acid (5), thus proving the involvement of novel lipoxygenase/lyase activity for the oxidation of C(16) fatty acids. PMID- 12633099 TI - Rhodium-catalyzed cross-coupling of allyl alcohols with aryl- and vinylboronic acids in ionic liquids. AB - [reaction: see text] The direct coupling of aryl- and vinylboronic acids with allylic alcohols has been achieved in ionic liquids using a rhodium catalyst. PMID- 12633100 TI - Palladium-catalyzed microwave-assisted amination of 1-bromonaphthalenes and 5- and 8-bromoquinolines. AB - [reaction: see text] 1-Aminonaphthalenes and 5- and 8-aminoquinolines were rapidly prepared from the respective aryl bromides in good yields by Pd-catalyzed aryl amination under microwave conditions. Consistent improvements in yields over those obtained under standard conditions were seen with quinoline substrates. In the cases where 5-bromo-8-cyanoquinoline was used as a substrate, no desired products were obtained under standard conditions with a number of different primary and secondary amines. However, microwave conditions provided the desired products in good to excellent yields. PMID- 12633101 TI - Alkenylthioimidoyl radicals: competition between beta-scission and cyclization to dihydrothiophen-2-ylidene-amines. AB - [reaction: see text] But-3-enylthioimidoyl radicals were shown by EPR spectroscopy and end product analysis to ring-close predominantly in the 5-exo mode with a rate constant of 2.4 x 10(4) s(-)(1) at 300 K to afford substituted dihydrothiophenylmethyl radicals. This ring closure was in competition with dissociation to but-3-enyl radicals and an isothiocyanate. The dissociation predominated at temperatures above ca. 300 K. PMID- 12633102 TI - Acid-catalyzed Grob fragmentation reactions of acetonides derived from terpenes. AB - [reaction: see text] Acetonides derived from different terpenes undergo Grob fragmentation by treatment with a catalytic amount of acid, triflic acid, or boron trifluoride, giving aldehydes containing a cyclopropane or cyclobutane ring with good yields and complete diastereoselectivity. The structure and the stereochemistry of the starting acetonide have a crucial influence on the reaction course. PMID- 12633103 TI - Aliphatic acetylenic homocoupling catalyzed by a novel combination of AgOTs-CuCl2 TMEDA and its application for the solid-phase synthesis of bis-benzo[b]furan linked 1,3-diynes. AB - [reaction: see text] A novel catalytic system of AgOTs-CuCl(2)-TMEDA is described for the homocoupling of aliphatic acetylenes on solid support. It is the first observation that Ag(I)'s activating triple bond could facilitate Cu(II)-mediated oxidative acetylenic homocoupling. This study provides an efficient way to synthesize a diversified symmetrical bis-benzo[b]furan-linked 1,3-diyne library on solid support. PMID- 12633104 TI - Synthesis of an F-H gambierol subunit using a C-glycoside-centered strategy. AB - [structure: see text] This manuscript describes our synthesis of the F-H subunit of gambierol. In addition to the synthesis of the tricycle, of note is an interesting protecting group influence on the generation of a C(23) C-glycoside as well as the use of ring-closing metathesis to generate a tetrasubstituted enol ether. PMID- 12633106 TI - Preparation of the 4-hydroxytryptamine scaffold via palladium-catalyzed cyclization: a practical and versatile synthesis of psilocin. AB - [reaction: see text] The 4-hydroxytryptamine scaffold of psilocin was successfully prepared via palladium-catalyzed cyclization of protected N-tert butoxycarbonyl-2-iodo-3-methoxyaniline and an appropriately substituted silyl acetylene. Removal of the protecting groups afforded psilocin in good yield. PMID- 12633105 TI - Stereospecific syntheses of 3'-deuterated pyrimidine nucleosides and their site specific incorporation into DNA. AB - [reaction: see text] 2'-Deoxy-3'-deutero pyrimidines have been synthesized in high yields and incorporated into deoxyoligonucleotides using standard phosphoramidite chemistry. A key synthetic step is a stereospecific reduction of 3'-keto nucleosides using sodium triacetoxyborodeuteride to give 3'-deuterated thymidine and 2'-deoxy uridine nucleosides. Conversion of the corresponding phorphoramidites 7a and 7b to 4-triazolo derivatives has, for the first time, enabled incorporation of 2'-deoxy-3'-deutero cytidine and 2'-deoxy-3'-deutero-5 methyl cytidine into oligonucleotides. PMID- 12633107 TI - N-sulfinyl beta-amino Weinreb amides: synthesis of enantiopure beta-amino carbonyl compounds. Asymmetric synthesis of (+)-sedridine and (-)-allosedridine. AB - [reaction: see text] N-Sulfinyl beta-amino Weinreb amides are prepared by condensation of sulfinimines with the potassium enolate of N-methoxy-N methylacetamide. These new chiral building blocks are useful for the asymmetric synthesis of beta-amino carbonyl compounds, as illustrated here by the concise enantioselective syntheses of sedum alkaloids (+)-sedridine and (-) allosedridine. PMID- 12633108 TI - Synthesis, chemistry, and dynamic NMR study of new atropisomeric 4-dialkylamino-5 chloro-1,2-dithiole-3-thiones. AB - [reaction: see text] Reaction of N-(2-phthalimidoethyl)-N-alkylisopropylamines and S(2)Cl(2) gave atropisomeric (by dynamic NMR) 4-N-(2-phthalimidoethyl)-N alkylamino-5-chloro-1,2-dithiole-3-thiones that quantitatively cycloadded to dimethyl or diethyl acetylenedicarboxylate to give a novel class of stable thioacid chlorides, which in turn reacted with 1 or 2 equiv of secondary amines to give thioamides. PMID- 12633110 TI - Interannular communication in the radical anions of bis-cyclooctatetraene systems. AB - [reaction: see text] The room-temperature potassium reduction of 1,2-bis cyclooctatetraeneoxypropane yields two different regio-spin isomer anion radicals in equilibrium (COT-O-C(Me)HCH(2)CH(2)-O-COT(*)(-) = (*)(-)COT-O-C(Me)HCH(2)CH(2) O-COT) that is shifted far to the right. The presence of the unreduced ring perturbs the spin density on the reduced ring. Addition of more electrons generates the diradical dianion ((*)(-)COT-O-C(Me)HCH(2)CH(2)-O-COT(*)(-)), and the anion radical on the secondary side splits the degeneracy of the psuedo-ortho protons of the anion radical on the primary side. PMID- 12633109 TI - Substrate specificity of NovM: implications for novobiocin biosynthesis and glycorandomization. AB - [reaction: see text] In an effort to expand the scope of natural product in vitro glycorandomization (IVG), the substrate specificity of NovM was investigated. A test of four aglycon analogues and over 40 nucleotide sugars revealed NovM has a surprisingly stringent substrate specificity and provided only three new "unnatural" natural products. On the basis of the determined substrate specificity, an alternative to the sugar nucleotide biosynthetic dogma and a cautionary note for the general applicability of IVG are introduced. PMID- 12633111 TI - Facile asymmetric synthesis of the core nuclei of xanthanolides, guaianolides, and eudesmanolides. AB - [reaction: see text] Bicyclic and tricyclic gamma-butyrolactones with 5,7-, 5,6,5 , 5,6,6-, or 5,7,5-fused ring systems, being found in xanthanolides, eudesmanolides, and guaianolides, were readily synthesized from methyl furan-2 carboxylic acid. Key steps were a copper(I)-catalyzed asymmetric cyclopropanation, Sakurai allylations, intramolecular ene reactions, and ring closing metathesis reactions. PMID- 12633112 TI - "N-acyliminium ion pool" as a heterodiene in [4 + 2] cycloaddition reaction. AB - [reaction: see text] An N-acyliminium ion pool was found to undergo cycloaddition reaction with a variety of dienophiles such as alkenes and alkynes. A concerted mechanism seems to be most likely for alkyl-substituted alkenes as suggested by the DFT calculations, whereas a stepwise mechanism plays the major role for aryl substituted alkenes. It is also noteworthy that the present study demonstrates the potential of the combination of the cation pool method and the micromixing in both mechanistic and synthetic aspects. PMID- 12633113 TI - Enantioselective cyanosilylation of ketones by a catalytic double-activation method employing chiral Lewis acid and achiral N-oxide catalysts. AB - [reaction: see text] Enantioselective addition of TMSCN to ketones is achieved by a catalytic double-activation method using 1a-Ti(IV) complex as the Lewis acid and achiral N-oxide 2 as the Lewis base to activate ketones and TMSCN, respectively. PMID- 12633114 TI - Novel class of tertiary phosphine ligands based on a phospha-adamantane framework and use in the Suzuki cross-coupling reactions of aryl halides under mild conditions. AB - [reaction: see text] A new class of sterically hindered phosphines based on a phospha-adamantane framework is described. Arylation or alkylation of the 1,3,5,7 tetramethyl-2,4,8-trioxa-6-phospha-adamantane system allows for the preparation of tertiary phosphines suitable for use in palladium-catalyzed cross-coupling reactions. For example, use of a catalytic system incorporating Pd(2)(dba)(3) and 1,3,5,7-tetramethyl-2,4,8-trioxa-6-phenyl-6-phospha-adamantane is shown to promote the Suzuki cross-coupling of aryl iodides, bromides, and activated chlorides with a variety of aryl boronic acids at room temperature in a few hours with high yields. PMID- 12633115 TI - Enantioselective synthesis of 15-epi-haterumalide NA methyl ester and revised structure of haterumalide NA. AB - [structure: see text] The enantioselective synthesis of the enantiomer of the haterumalide NA methyl ester, a cytotoxic macrolide from an Okinawan sponge, was achieved from the threitol derivative in 26 steps. The key steps are the stereoselective construction of a chloroolefin unit and the intramolecular Reformatsky-type reaction. This synthesis revised the absolute stereochemistry of haterumalide NA. PMID- 12633117 TI - Neurotransmission of cognition, part 2. Selective NRIs are smart drugs: exploiting regionally selective actions on both dopamine and norepinephrine to enhance cognition. AB - ISSUE: Selective norepinephrine reuptake inhibitors such as atomoxetine increase both dopamine and norepinephrine in frontal cortex and may thereby enhance cognitive functioning in attention-deficit/hyperactivity disorder. PMID- 12633118 TI - Estrogen administration does not reduce the rate of recurrence of affective psychosis after childbirth. AB - BACKGROUND: High rates of postpartum relapse occur in women with histories of bipolar or schizoaffective disorder. These relapses may be triggered by the postdelivery fall in circulating estrogen through alteration of central neurotransmitter (especially dopaminergic) systems. This study tested the hypothesis that estrogen administration after childbirth would prevent postpartum relapse and would alter dopamine receptor sensitivity. METHOD: Twenty-nine pregnant women with a Research Diagnostic Criteria diagnosis of hypomania (bipolar II), mania (bipolar I), or schizoaffective disorder participated in an open clinical trial. Three transdermal dose regimens of estrogen (17beta estradiol) were tested. Starting doses were 200 (N = 13), 400 (N = 3), and 800 (N = 13) micro g/day, beginning within 48 hours after delivery and reduced by one half every 4 days for a total of 12 days. On the fourth day after starting estradiol therapy (before relapse occurred), subjects participated in a neuroendocrine challenge test that measured the sensitivity of the central nervous system (tubero-infundibular) dopaminergic system (plasma prolactin and growth hormone responses to apomorphine). RESULTS: Estradiol at all dose regimens did not reduce the rate of relapse. However, of the 12 women who relapsed, those who had taken the highest dose of estradiol (800 micro g/day) needed less subsequent psychotropic medication (fewer chlorpromazine equivalents) and were discharged sooner than those who had taken either of the 2 lower doses. No differences in neuroendocrine responses to apomorphine were detected between women receiving the high-dose and the lower-dose regimens. CONCLUSION: The results do not support the hypothesis that a fall in circulating concentrations of estrogens precipitates relapse in subjects at risk of postpartum affective psychosis. The use of prophylactic estrogen in such circumstances is therefore highly questionable. PMID- 12633119 TI - Switching from depot antipsychotic drugs to olanzapine in patients with chronic schizophrenia. AB - BACKGROUND: Patients with chronic schizophrenia (DSM-IV criteria) often receive depot antipsychotic medications to assure longer administration and better compliance with their treatment regimen. This study evaluated whether patients stabilized on depot antipsychotic medication could be successfully transitioned to oral olanzapine. METHOD: In a 3-month open-label study, 26 clinically stable patients with schizophrenia taking depot antipsychotics for over 3 years were randomly assigned to continue on their current depot dose or to switch to oral olanzapine. Clinical ratings (Positive and Negative Syndrome Scale [PANSS], Global Assessment of Functioning [GAF] scale, and Clinical Global Impressions [CGI] scale) and side effect parameters (Abnormal Involuntary Movement Scale [AIMS], Barnes Akathisia Scale, AMDP-5 scale, vital signs, and weight) were obtained monthly. RESULTS: Oral olanzapine patients (N = 13) demonstrated significant clinical improvement over the depot control group (N = 13) from baseline to 3-month endpoint (PANSS total, p =.012; PANSS general, p =.068; PANSS negative, p =.098; CGI-Improvement, p =.007; CGI-Severity, p =.026; GAF, p =.015). Side effect rating scales showed no statistical differences between the 2 groups (AIMS, Barnes Akathisia Scale, AMDP-5, vital signs). The depot control group showed no statistical superiority in any measure except weight change (p =.0005). After 3 months, all olanzapine patients preferred olanzapine to their previous depot medications and chose to continue on olanzapine treatment. CONCLUSION: Clinicians may expect clinical improvement when switching chronically psychotic patients from traditional depot antipsychotic drugs to oral olanzapine. Switching may be completed within a 4-week period with relative compliance being maintained and patients preferring oral olanzapine to their previous depot medications. PMID- 12633120 TI - Can long-term treatment with antidepressant drugs worsen the course of depression? AB - BACKGROUND: The possibility that antidepressant drugs, while effectively treating depression, may worsen its course has received inadequate attention. METHOD: A review of the literature suggesting potential depressogenic effects of long-term treatment with antidepressant drugs was performed. A MEDLINE search was conducted using the keywords tolerance, sensitization, antidepressive agents, and switching. This was supplemented by a manual search of Index Medicus under the heading "antidepressant agents" and a manual search of the literature for articles pointing to paradoxical effects of antidepressants. RESULTS: A number of reported clinical findings point to the following possibilities: very unfavorable long-term outcome of major depression treated by pharmacologic means, paradoxical (depression-inducing) effects of antidepressant drugs in some patients with mood and anxiety disturbances, antidepressant-induced switching and cycle acceleration in bipolar disorder, occurrence of tolerance to the effects of antidepressants during long-term treatment, onset of resistance upon rechallenge with the same antidepressant drug in a few patients, and withdrawal syndromes following discontinuation of mood-elevating drugs. These phenomena in susceptible individuals may be explained on the basis of the oppositional model of tolerance. Continued drug treatment may recruit processes that oppose the initial acute effects of a drug and may result in loss of clinical effect. When drug treatment ends, these processes may operate unopposed, at least for some time, and increase vulnerability to relapse. CONCLUSION: The possibility that antidepressant drugs may worsen the course of depression needs to be tested, even though its scientific exploration is likely to encounter considerable methodological and ideological difficulties. The clinical implications of this hypothesis in depression are considerable. Antidepressant drugs are crucial in the treatment of major depressive episodes. However, appraisal of paradoxical effects that may occur in susceptible patients during long-term treatment may lead to more effective use of the drugs. PMID- 12633121 TI - A randomized placebo-controlled trial of risperidone for the treatment of aggression, agitation, and psychosis of dementia. AB - BACKGROUND: This randomized, double-blind, placebo-controlled trial examined the efficacy and safety of risperidone in the treatment of aggression, agitation, and psychosis in elderly nursing-home patients with dementia. METHOD: Elderly patients with a DSM-IV diagnosis of dementia of the Alzheimer's type, vascular dementia, or a combination of the 2 (i.e., mixed dementia) and significant aggressive behaviors were randomized to receive, for a period of 12 weeks, a flexible dose of either placebo or risperidone solution up to a maximum of 2 mg/day. Outcome measures were the Cohen-Mansfield Agitation Inventory (CMAI), the Behavioral Pathology in Alzheimer's Disease (BEHAVE-AD) rating scale, and the Clinical Global Impression of Severity (CGI-S) and of Change (CGI-C) scales. RESULTS: A total of 345 patients were randomized to treatment with risperidone or placebo, and 337 patients received at least one dose of study drug. The trial was completed by 67% of patients in the placebo group and 73% of patients in the risperidone group. The mean +/- SE dose of risperidone was 0.95 +/- 0.03 mg/day. The primary endpoint of the study, the difference from baseline to endpoint in CMAI total aggression score, showed a significant reduction in aggressive behavior for risperidone versus placebo (p <.001). A similar improvement was also seen for the CMAI total non-aggression subscale (p <.002) and for the BEHAVE-AD total (p <.001) and psychotic symptoms subscale (p =.004). At endpoint, the CGI-S and the CGI-C scores indicated a significantly greater improvement with risperidone compared with placebo (p <.001). Overall, 94% and 92% of the risperidone and placebo groups, respectively, reported at least 1 adverse event. Somnolence and urinary tract infection were more common with risperidone treatment, whereas agitation was more common with placebo. There was no significant difference in the number of patients who reported extrapyramidal symptoms between the risperidone (23%) and placebo (16%) groups. CONCLUSION: Treatment with low-dose (mean = 0.95 mg/day) risperidone resulted in significant improvement in aggression, agitation, and psychosis associated with dementia. PMID- 12633122 TI - Prophylactic efficacy of lithium versus carbamazepine in treatment-naive bipolar patients. AB - BACKGROUND: Alternatives to lithium for prophylactic treatment of patients with bipolar affective disorders are increasingly being advocated. However, trials comparing lithium with alternatives are scarce and often biased. METHOD: We studied 94 patients with at least 2 episodes of bipolar disorder (DSM-III-R) during the previous 3 years who were in remission at entry into the study. Treatment with lithium or carbamazepine had not exceeded a total of 6 months during their lifetime. Patients were randomly assigned to carbamazepine or lithium at entry into the 2-year double-blind study or during the acute index episode previous to entry into the study. No concurrent antipsychotics or antidepressants were allowed. RESULTS: On lithium treatment, 12/44 patients developed an episode, compared with 21/50 on carbamazepine treatment. Episodes on lithium treatment occurred almost exclusively during the first 3 months of the trial. Carbamazepine carried a constant risk of an episode of about 40% per year. Efficacy of lithium was superior to that of carbamazepine in patients with a (hypo)manic index episode that had not been treated with study drug during the index episode (p <.01) and also in patients with prior hypomanic but no manic episodes (p <.05). The proportion of patients who dropped out was slightly higher among those taking lithium (16/44) compared with those taking carbamazepine (13/50), resulting in 16/44 patients (36%) on lithium treatment completing the 2 years with no episode, compared with 16/50 (32%) on carbamazepine treatment. CONCLUSION: Lithium appears to be superior in prophylactic efficacy to carbamazepine in bipolar patients not previously treated with mood stabilizers. Our results should reinforce efforts to put and maintain such patients on treatment with lithium. PMID- 12633123 TI - How common is obsessive-compulsive disorder in a dermatology outpatient clinic? AB - BACKGROUND: This study was prompted by reports suggesting a high prevalence of unrecognized obsessive-compulsive disorder (OCD) in the dermatology clinic. METHOD: 92 consecutive dermatology referrals were screened for DSM-IV OCD using the Mini-International Neuropsychiatric Inverview (MINI), the Yale-Brown Obsessive Compulsive Scale (YBOCS), and the 5-item screening questionnaire from the International Council on OCD. Illness severity was rated on the YBOCS, and symptom profiles and dermatologic diagnoses were established for screen-positive cases. RESULTS: 18 patients (20%) qualified for a DSM-IV diagnosis of OCD, of whom 17 were previously undiagnosed. The range and type of OCD symptoms covered the normal clinical spectrum. Most patients had more than 1 symptom, and among obsessions (including somatic obsessions), checking, washing, and symmetry were common. The mean total YBOCS score was 16/40 (SD = 7.2), indicating moderate OCD, and 40% of the positive cases scored 16 or higher. Dermatologic diagnoses were various and did not seem to bear a direct relationship with the OCD. CONCLUSION: These results suggest that there is a high prevalence of clinically relevant OCD in the dermatology clinic. This is an area that merits attention with regard to better recognition and treatment for OCD sufferers. PMID- 12633124 TI - Past anabolic-androgenic steroid use among men admitted for substance abuse treatment: an underrecognized problem? AB - BACKGROUND: Recent reports suggest that anabolic-androgenic steroids (AAS) may cause mood disorders or dependence syndromes and may help to introduce some individuals to opioid abuse. At present, however, little is known about prior AAS use among men entering inpatient substance abuse treatment. METHOD: We assessed lifetime AAS use in 223 male substance abusers admitted to a substance abuse treatment unit primarily for treatment of alcohol, cocaine, and opioid dependence. Subjects reporting definite or possible AAS use were then asked to participate in a detailed semistructured interview that covered demographics, drug use history, and symptoms experienced during AAS use and withdrawal, and whether AAS use had helped introduce the subject to other classes of drugs. RESULTS: Twenty-nine men (13%) reported prior AAS use, but this history was documented on physicians' admission evaluations in only 4 cases. Among 88 men listing opioids as their drug of choice, 22 (25%) acknowledged AAS use, versus only 7 (5%) of the other 135 men (p <.001). Twenty-four (83%) of the 29 AAS users were interviewed in detail. Seven (29%) of the men interviewed, all with opioid dependence, reported that they first learned about opioids from friends at the gym and subsequently first obtained opioids from the same person who had sold them AAS. Eighteen (75%) of the men interviewed reported that AAS were the first drugs that they had ever self-administered by injection, 4 (17%) reported severe aggressiveness or violence during AAS use, 1 (4%) attempted suicide during AAS withdrawal, and 5 (21%) described a history of AAS dependence. CONCLUSION: Prior AAS use appears to be common but underrecognized among men entering inpatient substance abuse treatment, especially those with opioid dependence. AAS use may serve as a "gateway" to opioid abuse in some cases and may also cause morbidity in its own right. PMID- 12633125 TI - Perceptions and impact of bipolar disorder: how far have we really come? Results of the national depressive and manic-depressive association 2000 survey of individuals with bipolar disorder. AB - OBJECTIVE: To assess the experience of selected individuals living with bipolar disorder and compare this experience with that of a similar group of individuals sampled in 1992. METHOD: In June 2000, 4192 self-administered questionnaires were sent to National Depressive and Manic-Depressive Association chapters for distribution to support group participants diagnosed with bipolar disorder. By July 31, 2000, the first 600 completed surveys were analyzed. RESULTS: Over one third of respondents sought professional help within 1 year of the onset of symptoms. Unfortunately, 69% were misdiagnosed, with the most frequent misdiagnosis being unipolar depression. Those who were misdiagnosed consulted a mean of 4 physicians prior to receiving the correct diagnosis. Over one third waited 10 years or more before receiving an accurate diagnosis. Despite having underreported manic symptoms, more than half believe their physicians' lack of understanding of bipolar disorder prevented a correct diagnosis from being made earlier. In 2000, the respondents reported a greater negative impact of bipolar disorder on families, social relationships, and employment than did the respondents in 1992. Overall, respondents were satisfied with their current treatment, which often included medication, talk therapy, and support groups. Respondents who were highly satisfied with their treatment provider had a more positive outlook on their illness and their ability to cope with it. CONCLUSION: Individuals with bipolar disorder reported that the illness manifests itself early in life but that accurate diagnosis lags by many years. The illness exacts great hardships on the individual and the family and has a profoundly negative effect on careers. These findings are very similar to those reported nearly a decade ago. PMID- 12633126 TI - Posttraumatic stress disorder and its impact on the economic and health costs of motor vehicle accidents in South australia. AB - BACKGROUND: Motor vehicle accident studies thus far have focused primarily on psychiatric consequences and outcomes and medicolegal and treatment aspects, particularly of posttraumatic stress disorder (PTSD). This study aimed to determine the impact of motor vehicle accident-related psychiatric disorders on health and economic costs in quantitative terms. METHOD: Of the 3088 victims of motor vehicle accidents who made a claim through the State Insurance Commission, South Australia, between November 27, 1996, and March 23, 1999, 391 responded to the study and were assessed using the 28-item General Health Questionnaire, the PTSD Checklist-Civilian Version, and the Dissociative Experiences Scale. At the end of the study period, computerized cost records and accounting data on the health and economic costs incurred were obtained for each of the subjects. RESULTS: The total health and economic cost in Australian dollars for the 391 motor vehicle accident victims was A$6,369,519.52. At about 9 months after the accident, of the 391 subjects who replied to the questionnaires, 31% were identified as depressed and 62% as anxious, while 29% met criteria for PTSD. PTSD cases incurred significantly higher health care costs compared with non-PTSD cases (p <.001). Untreated PTSD cases incurred significantly higher economic losses compared with treated PTSD and non-PTSD cases (p <.05). CONCLUSION: The health and economic costs associated with motor vehicle accidents are enormous. Psychiatric morbidity among victims was high, and motor vehicle accident-related PTSD significantly contributed to increased overall health care and economic costs. PMID- 12633127 TI - Integrated family and individual therapy for bipolar disorder: results of a treatment development study. AB - BACKGROUND: Several studies have established the efficacy of psychosocial interventions as adjuncts to pharmacotherapy in the symptom maintenance of bipolar disorder. This study concerned a new psychosocial approach - integrated family and individual therapy (IFIT) - that synthesizes family psychoeducational sessions with individual sessions of interpersonal and social rhythm therapy. METHOD: Shortly after an acute illness episode, 30 bipolar patients (DSM-IV criteria) were assigned to open treatment with IFIT (up to 50 weekly sessions of family and individual therapy) and mood-stabilizing medications in the context of a treatment development study. Their outcomes over 1 year were compared with the outcomes of 70 patients from a previous trial who received standard community care, consisting of 2 family educational sessions, mood-stabilizing medications, and crisis management (CM). Patients in both samples were evaluated as to symptomatic functioning at entry into the project and then every 3 months for 1 year. RESULTS: Patients in IFIT had longer survival intervals (time without relapsing) than patients in CM. They also showed greater reductions in depressive symptoms over 1 year of treatment relative to their baseline levels. The results could not be explained by group differences in baseline symptoms or pharmacologic treatment regimens. CONCLUSION: Combining family and individual therapy with medication may protect episodic bipolar patients from early relapse and ongoing depressive symptoms. Further examination of this integrative model within randomized controlled trials is warranted. PMID- 12633128 TI - Risperidone augmentation decreases rapid eye movement sleep and decreases wake in treatment-resistant depressed patients. AB - BACKGROUND: The atypical antipsychotic agent risperidone has beneficial effects on mood in patients with schizophrenia. This study aimed to assess whether risperidone produced typical antidepressant-like effects in the polysomnogram of healthy subjects and in depressed patients unresponsive to antidepressant medication. METHOD: We measured the effect of a single dose of risperidone (1 mg) on the polysomnogram of 8 healthy volunteers in a placebo-controlled, double blind, crossover design. We also measured the effects of open-label risperidone treatment (0.5-1.0 mg daily) on the polysomnogram of 8 patients meeting DSM-IV criteria for major depressive disorder who had received therapeutic doses of an antidepressant with an unsatisfactory response. Sleep was recorded at baseline and following 2 weeks of risperidone addition. RESULTS: In the healthy volunteers, risperidone significantly decreased rapid eye movement (REM) sleep (p =.04). After 2 weeks of risperidone treatment, depressed patients had significantly less wake (p =.02) and REM sleep (p =.02). Scores on depression rating scales for the depressed patients showed a significant decline (p <.05). CONCLUSION: Risperidone administration decreases REM sleep in both healthy volunteers and medication-resistant depressed patients, an action characteristic of conventional antidepressant medication. In depressed patients, risperidone also decreased wake. The utility of risperidone as an augmentation agent in depressed patients merits controlled study. PMID- 12633129 TI - Lamotrigine in patients with bipolar disorder and cocaine dependence. AB - BACKGROUND: Bipolar disorder is associated with the highest substance abuse rates of any psychiatric illness. Therefore, treatments that stabilize mood and decrease drug use or cravings are of great interest. Open-label lamotrigine was examined in 30 outpatients with DSM-IV bipolar disorder and cocaine dependence. Lamotrigine was either added to existing medication regimens or used as monotherapy. METHOD: Lamotrigine was started at a dose of 25 mg/day (12.5 mg/day in those taking valproic acid) and titrated to a maximum dose of 300 mg/day. Subjects received a baseline evaluation including a structured clinical interview and weekly assessments for 12 weeks with the Hamilton Rating Scale for Depression (HAM-D), Young Mania Rating Scale (YMRS), Brief Psychiatric Rating Scale (BPRS), and Cocaine Craving Questionnaire (CCQ). At each appointment, a urine sample was obtained, and participants reported drug use during the previous week. The subjects consisted of 13 men and 17 women with cocaine dependence and bipolar I disorder (N = 22), bipolar II disorder (N = 7), or bipolar disorder not otherwise specified (N = 1), with a mean +/- SD age of 35.4 +/- 7.2 years. Data were analyzed using the last observation carried forward on all subjects who completed the baseline evaluation and at least 1 postbaseline assessment. RESULTS: Significant improvement was observed in HAM-D, YMRS, and BPRS scores (p < or =.02). Cravings also significantly decreased as measured by the CCQ (p <.001). Dollar amount spent on drugs decreased nonsignificantly. Lamotrigine was well tolerated, with no subjects discontinuing due to side effects. CONCLUSION: Lamotrigine treatment was well tolerated in this sample and associated with statistically significant improvement in mood and drug cravings but not drug use. The findings suggest that larger controlled trials of lamotrigine are needed in this population. PMID- 12633130 TI - Olfaction as a traumatic reminder in posttraumatic stress disorder: case reports and review. AB - BACKGROUND: Memory for odors that are associated with intense emotional experiences is often strongly engraved. Odors are claimed to be more closely connected to affect than other sensory experiences. They can serve as potent contextual cues for memory formation and emotional conditioning and can also serve as cues for olfactory flashbacks. Though trauma-related smells have long been noted by clinicians to be precipitants of traumatic memories in patients with posttraumatic stress disorder (PTSD), very few reports have been published that document this. CASE REPORTS: We review olfactory memories and olfactory flashbacks by presenting 3 cases that illustrate the role of olfaction in PTSD. In these cases olfaction is either a precipitant of PTSD symptoms or an important component of reexperiencing. DISCUSSION: In PTSD, seemingly nonspecific cues have the potential to precipitate traumatic memories with strong emotional components. These conditioned responses in PTSD are hypothesized to be mediated by specific brain areas, i.e., amygdala, hippocampus, and orbitofrontal cortex. Questions about smells as a traumatic reminder should be part of the routine assessment of intrusive memories in PTSD. In addition, smells may have the potential to provide cues to exposure situations in therapy or to facilitate de novo conditioning. PMID- 12633131 TI - A double-blind, placebo-controlled trial of the safety and efficacy of selegiline transdermal system without dietary restrictions in patients with major depressive disorder. AB - BACKGROUND: The monoamine oxidase (MAO) inhibitor selegiline has demonstrated antidepressant efficacy superior to placebo. A selegiline transdermal system (STS) has been developed with unique pharmacokinetic and pharmacodynamic properties that allow inhibition of central nervous system MAO-A and MAO-B enzymes while substantially avoiding inhibition of intestinal and liver MAO-A enzyme. This novel transdermal system provides targeted MAO inhibition without clinically significant increases in sensitivity to dietary tyramine. We investigated the safety and efficacy of STS in patients with major depressive disorder. METHOD: 365 outpatients 18 to 65 years old with a DSM-IV diagnosis of major depressive disorder were enrolled at 16 sites. A 17-item Hamilton Rating Scale for Depression (HAM-D-17) score of > or = 20 was required for entry. Patients were randomly assigned to receive either STS, 20 mg/20 cm(2), daily or placebo patch for up to 8 weeks. A tyramine-restricted diet was neither required nor advised. Efficacy, safety, and vital sign measures were obtained regularly. RESULTS: 289 patients were randomly assigned to treatment and received at least 1 on-therapy evaluation (STS, N = 145; placebo, N = 144). Although the effect size was modest, at endpoint, STS was statistically superior to placebo on the MADRS (p =.001) and HAM-D-28 (p =.039) ratings and showed a nonsignificant superiority on the HAM-D-17 (p =.069) and Clinical Global Impressions-Severity ratings (p <.055). Side effect profiles were similar for STS and placebo with the exception of application-site reaction, which was observed in 31.5% of STS patients and 15.1% of placebo-treated patients (p =.001). No significant differences were observed in blood pressure measures between treatment groups. CONCLUSION: Results from this double-blind, placebo-controlled clinical trial demonstrate that STS may have a modest, but statistically significant, antidepressant benefit compared with placebo and a similar safety profile compared with placebo in the absence of a tyramine-restricted diet. PMID- 12633132 TI - Internet pharmacy prescription and phentermine overdose. PMID- 12633133 TI - Ziprasidone augmentation of clozapine in 11 patients. PMID- 12633134 TI - Acute liver failure after administration of herbal tranquilizer kava-kava (Piper methysticum). PMID- 12633135 TI - Priapism associated with risperidone: a case report. PMID- 12633136 TI - Clozapine-induced rabbit syndrome. PMID- 12633137 TI - AMPA blockade may be the mechanism underlying the efficacy of topiramate in PTSD. PMID- 12633141 TI - Time to focus on the locus. PMID- 12633142 TI - Insights into the diagnosis and treatment of lysosomal storage diseases. AB - Lysosomal storage diseases (LSDs) are a group of genetic disorders that result from defective lysosomal metabolism or export of naturally occurring compounds. Signs and symptoms are variable both within and between disorders depending on the location and extent of storage. Many patients develop neurologic symptoms that become obvious from the newborn period to adulthood. Diagnosis of suspected patients can usually be made by measuring the activity of an enzyme or concentration of a metabolite in easily obtained tissue samples. Based on the considerable diagnostic experience of our laboratory, we aid the physician in selecting the appropriate tests to perform. Hematopoietic stem cell transplantation and enzyme replacement therapy are already available or in clinical trials for a number of LSDs. Early diagnosis is critical, especially since those patients who are treated before significant symptoms arise have the best chance for a positive outcome. PMID- 12633143 TI - Autosomal dominant inherited neuropathies with prominent sensory loss and mutilations: a review. AB - Hereditary sensory neuropathies (HSNs) are rare disorders characterized by progressive distal sensory loss, predominantly affecting the lower limbs. Foot ulcers, severe skin and bone infections, arthropathy, and amputations are frequent and feared complications. Occasionally, patients complain of spontaneous shooting or lancinating pain. Autonomic fibers can be affected to a variable degree. Patients with HSN can also have severe distal weakness, and some HSN variants have therefore been classified among the hereditary motor and sensory neuropathies (HMSNs). Molecular genetic studies of autosomal dominant inherited neuropathies with prominent sensory loss and ulceromutilating features have assigned the genetic loci for HMSN type 2B (Charcot-Marie-Tooth syndrome type 2B) and HSN type 1 to chromosomes 3q13-22 and 9q22.1-22.3, respectively. However, some families with HSN have been excluded for linkage to these loci, suggesting further genetic heterogeneity. Recently, disease-causing mutations in the SPTLC1 gene have been identified in patients with HSN type 1. In this review, we discuss the hallmark features associated with the distinct genetic subtypes of autosomal dominant inherited HSN and provide genotype-phenotype correlations. PMID- 12633144 TI - Neuronal loss is greater in the locus coeruleus than nucleus basalis and substantia nigra in Alzheimer and Parkinson diseases. AB - CONTEXT: Alzheimer disease (AD) and Parkinson disease (PD) are associated with neuronal degeneration in major subcortical nuclei, but few studies have examined the neuronal degeneration in these nuclei concurrently. OBJECTIVE: To identify clinical and pathological correlates of neuronal loss in the nucleus basalis (NB), locus coeruleus (LC), and substantia nigra pars compacta (SN) in AD and PD. DESIGN: The study sample comprised 86 cases with pathologically confirmed AD, 19 cases with PD, and 13 healthy elderly control subjects. The number of nucleolated neurons was counted in representative sections of the NB, LC, and SN. Effect sizes (ES) were computed to determine the standardized difference in cell counts relative to healthy controls. RESULTS: Cases of AD showed the greatest neuronal loss in the LC (ES = 3.16) followed by the NB (ES = 1.10), but variable loss in the SN (ES = 0.16). Cases of PD also showed the greatest neuronal loss in the LC (ES = 6.47), followed by the SN (ES = 2.58) and the NB (ES = 0.85). Significant correlations were found between the number of neurons in the NB and LC in PD (r = 0.54, P<.05), as well as AD (r = 0.24, P<.05). The duration of illness correlated with greater neuronal loss in the LC and NB in AD, and greater neuronal loss in the SN in PD. CONCLUSIONS: For both AD and PD the greatest neuronal loss was found in the LC. In AD, neuronal loss was most severe and best correlated with the duration of illness in the LC, rather than in NB as traditionally expected. Correlations between neuronal loss in the LC and NB (but not SN) in both PD and AD suggest that the former 2 nuclei may share common pathogenetic susceptibilities. Given the prominent loss of neurons in the LC, detection and treatment of noradrenergic deficiencies warrant attention in both AD and PD. PMID- 12633145 TI - Acute stroke increases QT dispersion in patients without known cardiac diseases. AB - BACKGROUND: Electrocardiographic changes are well known to appear with acute cerebrovascular events. OBJECTIVE: To investigate if QT dispersion (QTd) is increased in patients who have an acute stroke and if this increase could be related to lesion extent and/or localization. DESIGN: The study group consisted of 36 patients who had an acute stroke and no history or signs of cardiovascular disease. An age-matched control group (n = 19) free of cardiovascular disease was also included. Simultaneous 12-lead electrocardiograms (ECGs) were recorded within the first 24 hours (24h-ECG) and after 72 hours (72h-ECG) from stroke onset. QT dispersion was assessed both manually and automatically with assessors blinded to the clinical data. RESULTS: QT dispersion, corrected QTd, and automated QTd were significantly increased in the 24h-ECG compared with the 72h ECG (60 [range, 20-80] milliseconds vs 40 [range, 0-80] milliseconds, P<.005; mean [SD], 56 [19] vs 36 [21] milliseconds, P<.001; and 50 [range, 14-94] vs 34 [range, 0-84] milliseconds, P<.005, respectively). However, QTd in the 72h-ECG was similar to QTd in the control group. While in the 24h-ECG corrected QTd was significantly greater in patients with large infarcts and large hemorrhages (mean [SD], 70 [20] vs 51 [20] milliseconds, P<.05), in the 72h-ECG corrected QTd was greater in patients with right vs left-sided lesions (mean [SD], 39 [18] vs 24 [18] milliseconds, P<.05). CONCLUSIONS: QT dispersion is increased in the first 24 hours in patients with acute stroke and no cardiovascular disease compared with the control group. Although this finding seems to be related to the size of the lesion rather than to the localization or type of stroke, after 72 hours specific lesion localization could also influence the QTd. PMID- 12633146 TI - Acute infarction limited to the lenticular nucleus: clinical, etiologic, and topographic features. AB - BACKGROUND: Chronic diseases involving the putamen and globus pallidus induce parkinsonism and other movement disorders. Sensory and motor dysfunction from deep middle cerebral artery infarction is usually due to an involvement of the internal capsule. The clinical picture associated with isolated infarction of the lenticular nucleus is less well established. OBJECTIVE: To analyze clinical features, topographic correlations, and cause of purely lenticular ischemic infarction. PATIENTS AND METHODS: We reviewed 820 consecutive patients with deep hemispheral infarct included in the Lausanne Stroke Registry between 1986 and 1998 and selected those with isolated lenticular involvement on computed tomography or magnetic resonance imaging. RESULTS: Thirteen patients had pure lenticular infarction. All had faciobrachiocrural hemisyndrome, while none showed acute or delayed parkinsonism or abnormal movement. Nine patients had a lesion restricted to the putamen. Two of them had ataxic motor hemisyndrome and 7 had sensorimotor hemisyndrome (with ataxia in 4, left hemineglect in 1, and deep pain in the arm and leg in 1). Four patients had a lesion of putamen and globus pallidus externus. Three of them had motor hemisyndrome (with nonfluent aphasia in 2 and ataxia in 1) and 1 had ataxic sensorimotor hemisyndrome. All infarcts were in the territory of the medial perforating branches of the medial cerebral artery. Presumed cause of stroke was small-artery disease in 5, artery-to-artery embolism in 4, cardioembolism in 3 and undetermined in 1. CONCLUSIONS: Acute lenticular infarction induces mainly hemiparesis but no movement disorder. Associated sensory deficits, aphasia, and hemineglect underline clinically the function of the lenticular nucleus in connection with the prefrontal, temporal, and parietal cortices. PMID- 12633148 TI - Caspase gene expression in the brain as a function of the clinical progression of Alzheimer disease. AB - BACKGROUND: Caspase gene expression has previously been reported in terminal Alzheimer disease (AD) brain, but, currently, little is known about the temporal pattern of caspase gene expression relative to the onset and clinical progression of AD. OBJECTIVE: To derive a profile of caspase gene expression and proapoptotic indexes as a function of the clinical and neuropathologic progression of AD dementia. SETTING AND PATIENTS: Postmortem survey of nursing home patients characterized clinically by Clinical Dementia Rating (CDR) and neuropathologically by Consortium to Establish a Registry for Alzheimer's Disease criteria. DESIGN AND OUTCOME MEASURES: To assess messenger RNA expression of caspase-1, -2L, -2S, -3, -5, -6, -7, -8, and -9; apoptotic cell death by TUNEL assay; and poly (ADP-ribose) polymerase cleavage in postmortem brain tissue samples from cognitively normal (CDR 0), high risk of developing AD dementia (CDR 0.5), and severe dementia (CDR 5) cases. RESULTS: Compared with CDR 0 cases, elevated messenger RNA expression of caspase-1 and caspase-7 in the entorhinal cortex of CDR 0.5 cases coincided with increased poly (ADP-ribose) polymerase cleavage but not apoptotic cell injury. In the entorhinal cortex of CDR 5 cases, we found elevation of caspase-1, -2L, -3, -5, -6, -7, -8, and -9 and a greater than 4-fold increase in TUNEL-positive cells. Caspase messenger RNA expression was closely associated with neurofibrillary tangle and, to a lesser extent, neuritic plaque density. CONCLUSIONS: Proapoptotic mechanisms may be at play early in the onset of AD (before overt signs of apoptosis) and may be a conditional factor for later apoptotic cell injury or death. These data have relevance to potential therapeutic interventions for AD using selective caspase inhibitors. PMID- 12633147 TI - Association of life activities with cerebral blood flow in Alzheimer disease: implications for the cognitive reserve hypothesis. AB - BACKGROUND: Regional cerebral blood flow (CBF), a good indirect index of cerebral pathologic changes in Alzheimer disease (AD), is more severely reduced in patients with higher educational attainment and IQ when controlling for clinical severity. This has been interpreted as suggesting that cognitive reserve allows these patients to cope better with the pathologic changes in AD. OBJECTIVE: To evaluate whether premorbid engagement in various activities may also provide cognitive reserve. DESIGN: We evaluated intellectual, social, and physical activities in 9 patients with early AD and 16 healthy elderly controls who underwent brain H(2)(15)O positron emission tomography. In voxelwise multiple regression analyses that controlled for age and clinical severity, we investigated the association between education, estimated premorbid IQ, and activities, and CBF. RESULTS: In accordance with previous findings, we replicated an inverse association between education and CBF and IQ and CBF in patients with AD. In addition, there was a negative correlation between previous reported activity score and CBF in patients with AD. When both education and IQ were added as covariates in the same model, a higher activity score was still associated with more prominent CBF deficits. No significant associations were detected in the controls. CONCLUSIONS: At any given level of clinical disease severity, there is a greater degree of brain pathologic involvement in patients with AD who have more engagement in activities, even when education and IQ are taken into account. This may suggest that interindividual differences in lifestyle may affect cognitive reserve by partially mediating the relationship between brain damage and the clinical manifestation of AD. PMID- 12633149 TI - Treatment of Wilson disease with ammonium tetrathiomolybdate: III. Initial therapy in a total of 55 neurologically affected patients and follow-up with zinc therapy. AB - BACKGROUND: It is unclear what anticopper drug to use for patients with Wilson disease who present with neurologic manifestations because penicillamine often makes them neurologically worse and zinc is slow acting. OBJECTIVE: To evaluate the frequency of neurologic worsening and drug adverse effects with ammonium tetrathiomolybdate. DESIGN: Open-label study of 55 untreated patients (22 of them new) presenting with neurologic Wilson disease treated with tetrathiomolybdate varying from 120 to 410 mg/d for 8 weeks and then followed up for 3 years. Neurologic function was assessed with scored neurologic and speech tests. SETTING: A university hospital referral setting. PATIENTS: All untreated, newly diagnosed patients with neurologic Wilson disease. INTERVENTION: Treatment with tetrathiomolybdate. MAIN OUTCOME MEASURES: Neurologic function was evaluated by neurologic and speech examinations. Drug adverse effects were evaluated by complete blood cell counts and biochemical measures. RESULTS: Only 2 (4%) of 55 patients treated with tetrathiomolybdate showed neurologic deterioration, compared with an estimated 50% of penicillamine-treated patients. Five of the 22 new patients exhibited bone marrow suppression and 3 had aminotransferase elevations. These numbers are higher than in the original 33 patients and appear to be due primarily to a more rapid dose escalation. CONCLUSIONS: Tetrathiomolybdate shows excellent efficacy in patients with Wilson disease who present with neurologic manifestations. With rapid escalation of dose, adverse effects from bone marrow suppression or aminotransferase elevations can occur. PMID- 12633150 TI - Prevalence and characteristics of dementia in Parkinson disease: an 8-year prospective study. AB - BACKGROUND: Few longitudinal studies of dementia in Parkinson disease (PD) have been reported, and the proportion of patients with PD who eventually develop dementia is unknown. OBJECTIVE: To examine the 8-year prevalence, characteristics, and risk factors of dementia in patients with PD. METHODS: Patients were recruited from an epidemiological study of PD in the county of Rogaland, Norway, using explicit criteria for PD. Subjects with cognitive impairment at disease onset were excluded. A semistructured caregiver-based interview, cognitive rating scales, and neuropsychological tests were used to diagnose dementia according to criteria from the Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition at baseline and 4 and 8 years later. A population-based sample of 3295 subjects in the municipality of Odense, Denmark, was used as a comparison group and examined at baseline and after 2 and 5 years. RESULTS: We included 224 patients with PD (116 women). At baseline, 51 patients (26%) had dementia. Fifty-five patients died, and 10 refused follow-up without their dementia status known. Forty-three and 28 new cases of dementia were identified at the 4- and 8-year evaluations, respectively. The 4-year prevalence of dementia in PD was nearly 3 times higher than in the non-PD group. The 8-year prevalence in PD was 78.2% (95% confidence interval [CI], 71.1-84.0). Risk factors for dementia were hallucinations before baseline (odds ratio [OR] = 3.1; 95% CI, 1.6-6.2) and akinetic-dominant or mixed tremor/akinetic PD (OR = 3.3; 95% CI, 1.2-8.5). CONCLUSIONS: More than three quarters of this representative PD cohort developed dementia during the 8-year study period. Early hallucinations and akinetic-dominant PD were associated with an increased risk of dementia. PMID- 12633151 TI - White matter structural integrity in healthy aging adults and patients with Alzheimer disease: a magnetic resonance imaging study. AB - BACKGROUND: Imaging and postmortem studies suggest that frontal lobe white matter (FLWM) volume expands until about the age of 44.6 years and then declines. Postmortem evidence indicates that the structural integrity of myelin sheaths deteriorates during normal aging, especially in late myelinating regions such as the frontal lobes. OBJECTIVES: To assess the integrity of FLWM by magnetic resonance imaging and, thus, to provide an important index of brain aging and its relationship to Alzheimer disease (AD). DESIGN: Cross-sectional study. SETTING: Two metropolitan university hospitals and AD research centers. PARTICIPANTS: Two hundred fifty-two healthy adults (127 men and 125 women), aged 19 to 82 years, and 34 subjects with AD (16 men and 18 women), aged 59 to 85 years. MAIN OUTCOME MEASURE: Calculated transverse relaxation rate (R( 2)) of the FLWM (an indirect measure of the structural integrity of white matter). RESULTS: As expected from prior imaging data on FLWM volume, the quadratic function best represented the relationship between age and the FLWM R(2) (P<.001). In healthy individuals, the FLWM R(2) increased until the age of 38 years and then declined markedly with age. The R( 2) of subjects with AD was significantly lower than that of a group of healthy control subjects who were of similar age and sex (P<.001). CONCLUSIONS: The R(2) changes in white matter suggest that the healthy adult brain is in a constant state of change, roughly defined as periods of maturation continuing into middle age followed by progressive loss of myelin integrity. Clinically diagnosed AD is associated with more severe myelin breakdown. Noninvasive measures, such as the determination of the R(2), may have the potential to track prospectively the trajectory of deteriorating white matter integrity during normal aging and the development of AD and, thus, may be a useful marker for medication development aimed at the prevention of AD. PMID- 12633152 TI - Cortical myoclonus during lithium exposure. AB - BACKGROUND: Myoclonus can occur in association with lithium therapy at toxic and therapeutic dosages, and can be a predominant and disabling adverse effect. Moreover, myoclonus has been reported when lithium has been combined with cyclic antidepressants and with the neuroleptic clozapine. Although clinical case reports exist, no electrophysiologic data are available that provide a source or a neurophysiological mechanism for the myoclonus seen in lithium therapy. OBJECTIVE: To describe the electrophysiologic characteristics and source of the myoclonus associated with lithium therapy. DESIGN AND METHODS: We retrospectively analyzed 5 cases of myoclonus during lithium therapy. We reviewed the clinical features and results of previous electrophysiologic testing. Four patients received lithium monotherapy; and 1, sertraline hydrochloride and nefazodone hydrochloride in addition to lithium. The electrophysiologic data that had been gathered included multichannel surface electromyographic (EMG) recordings with simultaneous electroencephalography (EEG), somatosensory evoked potentials, and elicitation of long-latency EMG reflexes to median and digital nerve stimulation. RESULTS: All 5 patients showed multifocal action myoclonus without reflex activation and only rare occurrence at rest. In each case, back-averaging created a focal EEG transient over the contralateral sensorimotor area preceding the myoclonus EMG discharge. In 2 of the patients receiving lithium monotherapy, the therapy was discontinued and the myoclonus disappeared. CONCLUSIONS: Lithium, by itself, can be associated with prominent clinical myoclonus, short-duration (<50 millisecond) myoclonus EMG discharges and cortical action myoclonus without the presence of epileptiform abnormalities on the routine EEG. This myoclonus is different from the most common form that is well documented to occur with tricyclic antidepressant therapy by clinical and electrophysiologic means. PMID- 12633153 TI - Rest tremor in patients with essential tremor: prevalence, clinical correlates, and electrophysiologic characteristics. AB - BACKGROUND: Isolated rest tremor, which is observed in some patients with essential tremor (ET), poses a diagnostic challenge. The phenomenon has been examined in few studies and is poorly understood. OBJECTIVES: To determine the prevalence and study the clinical correlates of rest tremor in ET and to examine the electrophysiologic features in a subgroup of patients. METHODS: Sixty-four patients with ET cared for at a tertiary referral center underwent neurologic examination. Five of 12 patients with rest tremor also underwent quantitative computerized tremor analysis using accelerometry and handwritten spiral analysis. RESULTS: Twelve of 64 patients with ET had rest tremor (prevalence, 18.8%; 95% confidence interval, 9.2%-28.4%). Compared with the 52 patients with ET without rest tremor, these 12 had disease of longer duration and greater severity. Also, their ET was more widely disseminated, as evidenced by a larger proportion with head tremor. None had clinical signs of bradykinesia or rigidity. The 5 patients with rest tremor who underwent electrophysiologic study had electrophysiologic features consistent with parkinsonism (eg, slow spiral speed and increased decrement of spiral speed with radius). CONCLUSIONS: In our sample, 1 in 5 patients with ET had rest tremor. The tremor was associated with disease that was more severe, more disseminated, and of longer duration. Some of these patients had electrophysiologic features consistent with parkinsonism. The basis for the rest tremor could be basal ganglia involvement, raising the possibility that the pathologic process responsible for ET may extend to these structures. PMID- 12633154 TI - Bilateral high-frequency synchronous discharges: a new form of tremor in humans. AB - OBJECTIVES: To report bilateral high-frequency synchronous discharges in a patient with a sporadic form of olivopontocerebellar atrophy; to investigate the electromyographic pattern, the coherence and cospectral density across limbs, and the resetting effects of electrical stimulation over the posterior fossa; and to highlight the clinical, electrophysiologic, and radiologic features of this new form of tremor associated with posterior fossa disorders. DESIGN: Case study of a patient clinically exhibiting a sporadic form of olivopontocerebellar atrophy associated with cerebellar and brainstem atrophy. SETTING: Research unit, university hospital. MAIN OUTCOME MEASURES: Electromyographic studies, coherence and cospectral density analysis, and calculation of a resetting index based on the timing of measured bursts and predicted bursts for an electrical stimulus given over the posterior fossa at increasing delays. RESULTS: Surface electromyographic recordings in forearm muscles revealed a low-frequency postural tremor in the upper limbs, with episodes of highly coherent tremor at a frequency of 14 Hz. Squared coherence and cospectral density was strong between agonist and antagonist muscles in the left and right upper limbs and across limbs for the high-frequency discharges. Electrical stimulation over the posterior fossa reset the explosive high-frequency bursts. The resetting index was 0.82 Hz. CONCLUSIONS: Our results show that bilateral high-frequency synchronous discharges may be associated with the sporadic form of olivopontocerebellar atrophy. Bilateral coherent bursting and resetting of this explosive postural tremor following electrical stimulation over the posterior fossa strongly suggest that the brainstem plays a key role in the network involved in the genesis of rhythmic bursts. We suggest that the high-frequency discharges are due to repetitive discharges in the reverberating cerebello-precerebellar circuits. PMID- 12633155 TI - The pathogenesis and treatment of optic disc swelling in neurosarcoidosis: a unique therapeutic response to infliximab. AB - OBJECTIVE: To review the pathogenesis and treatment of optic disc swelling in neurosarcoidosis, including a novel therapeutic response to infliximab. DESIGN AND SETTING: Case reports from an inpatient neurology service. PATIENTS: A 35 year-old woman presented with headache, chronic visual loss, papilledema, and optic atrophy, characteristic of chronic intracranial hypertension. Magnetic resonance imaging showed bifrontal cerebral edema with en plaque frontal pachymeningeal enhancement. Her visual loss progressed despite conventional therapies. The use of the tumor necrosis factor alpha antagonist infliximab maintained functional vision in her right eye. A 57-year-old woman presented with bilateral, subacute, painful visual loss and unilateral papillitis consistent with optic neuritis. Her visual loss responded rapidly to intravenous corticosteroids. The funduscopic examination findings in both patients prompted further clinical investigation, culminating in the diagnosis of neurosarcoidosis. CONCLUSION: Understanding the multiple etiologic mechanisms that produce optic disc swelling in sarcoidosis can help neurologists tailor treatment for patients with neurosarcoidosis who present with this symptom. PMID- 12633156 TI - Late-developing cerebral arteropathy after pyogenic meningitis. AB - BACKGROUND: Although vasculopathy is a recognized complication during acute meningitis, to our knowledge, no previous reports have been published of this phenomenon developing months after successful treatment. OBJECTIVE: To report a unique case of a late-developing vasculopathy after pyogenic meningitis in an adult. REPORT OF A CASE: A 51-year-old woman was seen with severe headache 2 months after treatment of Haemophilus influenzae type C meningitis. Initial arteriography showed no abnormality; a second arteriogram showed progressive multifocal intracranial stenosis affecting mainly the internal carotid arteries. Findings from pathologic examination disclosed diffuse collagenosis consistent with chronic vascular injury from meningitis. The arterial lesions stabilized, and the patient remained asymptomatic. CONCLUSION: Progressive intracranial arterial stenosis can evolve months after meningitis and should be added to the list of recognized vascular complications. PMID- 12633157 TI - A case of multiple brain infarctions associated with Erysipelothrix rhusiopathiae endocarditis. AB - A 63-year-old woman was admitted to our hospital because of fever and altered mentality. Brain magnetic resonance imaging showed multiple infarctions at the basal ganglia, cerebellum, and subcortical white matter with petechial hemorrhage, which was more easily seen on gradient echo images. Erysipelothrix rhusiopathiae was cultured from her blood, and echocardiography showed septic vegetations in the mitral valve. She recovered fully after 6 weeks of appropriate antibiotic treatment. PMID- 12633158 TI - Right orbitofrontal tumor with pedophilia symptom and constructional apraxia sign. AB - BACKGROUND: Orbitofrontal abnormalities are associated with poor impulse control, altered sexual behavior, and sociopathy. OBJECTIVE: To describe a patient with acquired pedophilia and a right orbitofrontal tumor who was unable to inhibit sexual urges despite preserved moral knowledge. DESIGN: Case report. RESULTS: The patient displayed impulsive sexual behavior with pedophilia, marked constructional apraxia, and agraphia. The behavioral symptoms and constructional deficits, including agraphia, resolved following tumor resection. CONCLUSIONS: For patients with acquired sociopathy and paraphilia, an orbitofrontal localization requires consideration. This case further illustrates that constructional apraxia can arise from right prefrontal lobe dysfunction. Agraphia may represent a manifestation of constructional apraxia in the absence of aphasia and ideomotor apraxia. PMID- 12633159 TI - Air myelopathy following cervical laminectomy and fusion. PMID- 12633160 TI - Using extirpations to understand the human motor cortex: Horsley, Foerster, and Bucy. AB - During the last part of the 19th century and through the middle of the 20th century, surgeons sometimes extirpated parts of the human motor cortex to control abnormal involuntary movements. This procedure can be traced directly to Victor Horsley, who pioneered the first successful surgery of this type in 1886. Although many neurosurgeons followed Horsley's lead in performing this procedure, few used their results, as he did, to formulate concepts on the role of the motor cortex in movement control. Otfrid Foerster and Paul Bucy were the principal exceptions. We reviewed the surgical procedures these 3 notable neurosurgeons performed on the motor cortex and the hypotheses they subsequently developed on the functions of the motor cortex. We also evaluated these writings relative to contemporary views of motor cortex function. PMID- 12633161 TI - Accuracy of diagnosis in dementia with Lewy bodies. PMID- 12633162 TI - Neuropathologic changes in Parkinson disease with late onset of dementia. PMID- 12633164 TI - Medicine on the Lewis and Clark Trail: exhibit explores expedition's medical adventures. PMID- 12633165 TI - CDC report on environmental toxins: some progress, some concerns. PMID- 12633170 TI - Definitions of the metabolic syndrome. PMID- 12633172 TI - Definitions of the metabolic syndrome. PMID- 12633173 TI - Definitions of the metabolic syndrome. PMID- 12633174 TI - Blood transfusions and mortality among critically ill patients. PMID- 12633175 TI - Blood transfusions and mortality among critically ill patients. PMID- 12633176 TI - Blood transfusions and mortality among critically ill patients. PMID- 12633177 TI - From the Centers for Disease Control and Prevention. Facilitating influenza and pneumococcal vaccination through standing orders programs. PMID- 12633178 TI - Medical research in pedophilia. PMID- 12633181 TI - Dangers to elderly pedestrians at crosswalks. PMID- 12633183 TI - Dangers to elderly pedestrians at crosswalks. PMID- 12633185 TI - Hepatitis C virus and atherosclerosis in patients with type 2 diabetes. PMID- 12633184 TI - Hyperglycemia in acute illness. PMID- 12633187 TI - Association of pediatric dental caries with passive smoking. AB - CONTEXT: Dental decay is the most common chronic disease of children and it disproportionately affects those living in poverty, but the reasons for this are not clear. Passive smoking may be a modifiable risk factor for dental caries. OBJECTIVE: To examine the relationship between dental caries and serum cotinine levels. DESIGN, SETTING, AND PARTICIPANTS: Cross-sectional data from the Third National Health and Nutrition Examination Survey (1988-1994) of 3531 children aged 4 to 11 years, who had had both dental examinations and a serum cotinine level measurement. MAIN OUTCOME MEASURES: Passive smoking defined as serum cotinine levels of 0.2 to 10 ng/mL and caries defined as decayed (unfilled) or filled tooth surfaces. RESULTS: Twenty-five percent of the children had at least 1 unfilled decayed tooth surface and 33% had at least 1 filled surface. Fifty three percent had cotinine levels consistent with passive smoking. Elevated cotinine level was significantly associated with both decayed (odds ratio [OR], 2.1; 95% confidence interval [CI], 1.5-2.9) and filled (OR, 1.4; 95% CI, 1.1-1.8) tooth surfaces in deciduous but not in permanent teeth. This relationship persisted after adjusting for age, sex, race, family income, geographic region, frequency of dental visits, and blood lead level. For dental caries in deciduous teeth, the adjusted OR was 1.8 (95% CI, 1.2-2.7) for the risk of decayed surfaces and 1.4 (95% CI, 1.1-2.0) for filled surfaces. We estimated the population attributable risk from passive smoking to be 27% for decayed and 14% for filled tooth surfaces. CONCLUSIONS: There is an association between environmental tobacco smoke and risk of caries among children. Reduction of passive smoking is important not only for the prevention of many medical problems, but also for the promotion of children's dental health. PMID- 12633186 TI - Plasma homocysteine and risk for congestive heart failure in adults without prior myocardial infarction. AB - CONTEXT: Elevated plasma homocysteine levels are associated with increased risk of vascular disease. It is unclear whether elevated homocysteine levels are a risk factor for congestive heart failure (CHF). OBJECTIVE: To study prospectively the association between nonfasting plasma homocysteine and incidence of CHF. DESIGN, SETTING, AND PARTICIPANTS: Community-based prospective cohort study of 2491 adults (mean age 72 years, 1547 women) who participated in the Framingham Heart Study during the 1979-1982 and 1986-1990 examinations and were free of CHF or prior myocardial infarction (recognized or unrecognized) at baseline. MAIN OUTCOME MEASURE: Incidence of a first episode of CHF during an 8-year follow-up period. RESULTS: During follow-up, 156 subjects (88 women) developed CHF. In multivariable analyses controlling for established risk factors for CHF including the occurrence of myocardial infarction (recognized or unrecognized) during follow-up, plasma homocysteine levels higher than the sex-specific median value were associated with an adjusted hazards ratio for heart failure of 1.93 in women (95% confidence interval, 1.19-3.14) and 1.84 in men (95% confidence interval, 1.06-3.17). The relation of plasma homocysteine levels to CHF risk was more continuous in women than in men. In analyses restricted to participants without any manifestation of coronary heart disease at baseline, the association of plasma homocysteine levels with risk of CHF was maintained in men and women. CONCLUSIONS: An increased plasma homocysteine level independently predicts risk of the development of CHF in adults without prior myocardial infarction. Additional investigations are warranted to confirm these findings. PMID- 12633188 TI - Population-based study of chlamydial infection in China: a hidden epidemic. AB - CONTEXT: Sexually transmitted diseases are increasing rapidly in China. Surveillance data imperfectly indicate current prevalence and risk factors. OBJECTIVES: To estimate the prevalence of genital chlamydial and gonococcal infections and to describe patterns of infection by subgroup and behavioral patterns. DESIGN, SETTING, AND PARTICIPANTS: A national stratified probability sample of 3426 Chinese individuals (1738 women and 1688 men) aged 20 to 64 years, who were interviewed between August 1999 and August 2000, completed a computer administered survey, and provided a urine specimen (69% total participation rate). MAIN OUTCOME MEASURE: Positive test result for chlamydial or gonococcal infections. RESULTS: The overall prevalence per 100 population of chlamydial infection was 2.6 (95% confidence interval [CI], 1.6-4.1) for women and 2.1 (95% CI, 1.3-3.3) for men. For gonococcal infection, the overall prevalence per 100 population was 0.08 (95% CI, 0.02-0.4) for women and 0.02 (95% CI, 0.005-0.1) for men. Risk factors for chlamydial infection among men aged 20 to 44 years were unprotected sex with a commercial sex worker (odds ratio [OR], 8.24; 95% CI, 3.51 19.35), less education (OR, 7.20; 95% CI, 2.31-22.37), and recent sex with their spouse or other steady partner (OR, 7.73; 95% CI, 2.70-22.10). Among women aged 20 to 44 years, risk factors for chlamydial infection were having less education (OR, 2.82; 95% CI, 1.01-7.91) and living in a city (OR, 3.46; 95% CI, 1.67-7.18) or along the southern coast (OR, 2.16; 95% CI, 1.29-3.63) and having a spouse or other steady sexual partner who earned a high income (OR, 2.85; 95% CI, 1.11 7.29), who socialized often (OR, 2.79; 95% CI, 1.08-7.19), or who traveled less than 1 week per year (OR, 5.40; 95% CI, 1.44-20.3). CONCLUSIONS: The prevalence of chlamydial infection in China is substantial. The patterns of infection suggest potential avenues for intervention. PMID- 12633189 TI - Efficacy of selected hand hygiene agents used to remove Bacillus atrophaeus (a surrogate of Bacillus anthracis) from contaminated hands. AB - CONTEXT: The intentional use of Bacillus anthracis transmitted via the US mail in October-November 2001 resulted in 22 people developing inhalation or cutaneous anthrax. Glove use with handwashing prior to and after contact with potential contaminated environmental surfaces and cutaneous lesions has been recommended. However, only limited data are available on the susceptibility of B anthracis to antiseptics. OBJECTIVE: To evaluate the efficacy of several hand antiseptics (interventions) and soap and water (control) against Bacillus atrophaeus, a surrogate of B anthracis. DESIGN, SETTING, AND PARTICIPANTS: Challenge study conducted among healthy adult volunteers, using the Standard Test Method for Evaluation of the Effectiveness of Health Care Professional Handwash Formulations (American Society for Testing and Materials E 1174-94) to determine the efficacy of various hand hygiene products at wash times of 10, 30, and 60 seconds. Volunteers were excluded if they had eczema, psoriasis, or other chronic skin conditions; nonintact skin; or allergies to any study agent. Study agents were a waterless rub containing 61% ethyl alcohol, a 2% chlorhexidine gluconate preparation, and an antibacterial microfiber towel that releases hypochlorite. A nonantimicrobial soap was used as a control. MAIN OUTCOME MEASURE: Reduction of B atrophaeus spores (log10 CFU/mL) on contaminated hands. RESULTS: Washes of 10, 30, and 60 seconds with either soap and water or 2% chlorhexidine gluconate eliminated 1.5 to 2.0 log10 CFUs/mL of B atrophaeus spores at wash 3. Mean reductions (95% confidence intervals) with 10-, 30-, and 60-second washes with soap and water were 2.4 (2.2-2.5), 2.3 (2.2-2.4), and 2.1 (1.9-2.4) log(10) CFUs/mL, respectively; and with 2% chlorhexidine gluconate, 2.1 (2.0-2.3), 1.8 (1.5-2.0), and 1.7 (1.5-1.9) log10 CFUs/mL, respectively. Handwashing with chlorine-containing towels was increasingly effective as the wipe time increased; reductions at 10, 30, and 60 seconds were 1.3 (1.1-1.5), 1.6 (1.2-2.0), and 2.2 (2.1-2.2) log10 CFUs/mL, respectively. A waterless rub containing 61% ethyl alcohol was ineffective in eliminating B atrophaeus spores at all times tested (0 [-0.1 to 0.1], -0.2 [-0.3 to -0.1], and 0 [-0.2 to 0.2] log10 CFUs/mL). CONCLUSIONS: In this evaluation of hand hygiene agents, handwashing with soap and water, 2% chlorhexidine gluconate, or chlorine-containing towels reduced the amount of B atrophaeus spore contamination, whereas use of a waterless rub containing ethyl alcohol was not effective in removing spores. PMID- 12633190 TI - Central challenges facing the national clinical research enterprise. AB - Medical scientists and public health policy makers are increasingly concerned that the scientific discoveries of the past generation are failing to be translated efficiently into tangible human benefit. This concern has generated several initiatives, including the Clinical Research Roundtable at the Institute of Medicine, which first convened in June 2000. Representatives from a diverse group of stakeholders in the nation's clinical research enterprise have collaborated to address the issues it faces. The context of clinical research is increasingly encumbered by high costs, slow results, lack of funding, regulatory burdens, fragmented infrastructure, incompatible databases, and a shortage of qualified investigators and willing participants. These factors have contributed to 2 major obstacles, or translational blocks: impeding the translation of basic science discoveries into clinical studies and of clinical studies into medical practice and health decision making in systems of care. Considering data from across the entire health care system, it has become clear that these 2 translational blocks can be removed only by the collaborative efforts of multiple system stakeholders. The goal of this article is to articulate the 4 central challenges facing clinical research at present--public participation, information systems, workforce training, and funding; to make recommendations about how they might be addressed by particular stakeholders; and to invite a broader, participatory dialogue with a view to improving the overall performance of the US clinical research enterprise. PMID- 12633191 TI - Colorectal cancer screening: scientific review. AB - CONTEXT: Screening for colorectal cancer clearly reduces colorectal cancer mortality, yet many eligible adults remain unscreened. Several screening tests are available, and various professional organizations have differing recommendations on which screening test to use. Clinicians are challenged to ensure that eligible patients undergo colorectal cancer screening and to guide patients in choosing what tests to receive. OBJECTIVE: To critically assess the evidence for use of the available colorectal cancer screening tests, including fecal occult blood tests, sigmoidoscopy, colonoscopy, double-contrast barium enema, and newer tests, such as virtual colonoscopy and stool-based molecular screening. DATA SOURCES: All relevant English-language articles were identified using PubMed (January 1966-August 2002), published meta-analyses, reference lists of key articles, and expert consultation. DATA EXTRACTION: Studies that evaluated colorectal cancer screening in healthy individuals and assessed clinical outcomes were included. Evidence from randomized controlled trials was considered to be of highest quality, followed by observational evidence. Diagnostic accuracy studies were evaluated when randomized controlled trials and observational studies were not available or did not provide adequate evidence. Studies were excluded if they did not evaluate colorectal screening tests and if they did not evaluate average risk individuals. DATA SYNTHESIS: Randomized controlled trials have shown that fecal occult blood testing can reduce colorectal cancer incidence and mortality. Case-control studies have shown that sigmoidoscopy is associated with a reduction in mortality, and observational studies suggest colonoscopy is effective as well. Combining fecal occult blood testing and sigmoidoscopy may decrease mortality and can increase diagnostic yield. CONCLUSION: The recommendation that all men and women aged 50 years or older undergo screening for colorectal cancer is supported by a large body of direct and indirect evidence. At present, the available evidence does not currently support choosing one test over another. PMID- 12633192 TI - Colorectal cancer screening: clinical applications. AB - Screening for colorectal cancer reduces mortality in individuals aged 50 years or older. A number of screening tests, including fecal occult blood tests, sigmoidoscopy, double-contrast barium enema, and colonoscopy, are recommended by professional organizations for colorectal cancer screening, yet the rates of colorectal cancer screening remain low. Questions regarding the quality of evidence for each screening test, whether screening for individuals at higher risk should be modified, the availability of the tests, and cost-effectiveness are addressed. Many potential barriers to colorectal cancer screening exist for the patient and the physician. Strategies to increase compliance for colorectal cancer screening are proposed. PMID- 12633193 TI - Hidden epidemic of sexually transmitted diseases in China: crisis and opportunity. PMID- 12633194 TI - Translating biomedical research to the bedside: a national crisis and a call to action. PMID- 12633195 TI - Conjoined twins. PMID- 12633196 TI - From the Centers for Disease Control and Prevention. Human rabies--Iowa, 2002. PMID- 12633197 TI - From the Centers for Disease Control and Prevention. Hypothermia related deaths- Philadelphia, 2001, and United States, 1999. PMID- 12633198 TI - JAMA patient page. Colon cancer screening. PMID- 12633199 TI - A grading system for nasal dorsal deformities. AB - There is no uniform grading system for nasal dorsal deformities currently in general use among surgeons who perform rhinoplasty. Given the popularity of this procedure among both the general public and surgeons, it is time that there was a uniform system describing dorsal deformities. Such a system has value in the education of students of rhinology and cosmetic nasal surgery. We have developed one such system, and applied it to 100 cases. In all cases it accurately describes the major pathological conditions of the dorsum, if present, as noted on physical examination. We have found application of this system to be facile. PMID- 12633200 TI - Skin rejuvenation regimens: a profilometry and histopathologic study. AB - OBJECTIVE: To quantitatively examine the effects of skin rejuvenation regimens in treating photodamaged skin. METHODS: Fourteen patients with photodamaged skin were considered for analysis. Skin rejuvenation regimens were as follows: (1) 10 weeks of treatment with topical 0.05% tretinoin emollient cream, (2) 10 weeks of treatment with 0.05% tretinoin emollient cream and topical ascorbic acid lotion, (3) 6 superficial trichloroacetic acid peels, and (4) a combination of the topical treatments and superficial peels. Comparisons of the treatments were analyzed using profilometry and histologic findings. RESULTS: Profilometry analysis provided quantification of the changes from each treatment group and among the treatment programs. Each group showed improvements from baseline. Trichloroacetic acid peels combined with application of the topical products improved skin topography to a greater extent than the less aggressive regimens. Histologic changes correlated well with the skin replica findings. CONCLUSIONS: A 10-week skin rejuvenation regimen objectively improved photodamaged facial skin. Significant changes are noted when combining topical treatments with superficial peels. Hence, use of a combination of trichloroacetic acid peels, 0.05% tretinoin emollient cream, and ascorbic acid lotions is well tolerated and superior to either component alone as part of a comprehensive skin care and sun protection program. PMID- 12633201 TI - Anatomical guides to precisely localize the frontal branch of the facial nerve. AB - OBJECTIVE: To define the relationship between the frontal branch of the facial nerve and a series of blood vessels that are encountered during endoscopic forehead procedures. DESIGN: Anatomical study using 6 fresh cadavers (12 sides). RESULTS: In 11 of 12 dissected specimens, the blood vessels in the temporal region were found to lie within 2 mm of the frontal branch of the facial nerve. CONCLUSIONS: We believe that a series of veins encountered during endoscopic forehead procedures provide the surgeon with the ability to identify the precise location of the frontal branch of the facial nerve. During endoscopic surgery, these vessels are found in a plane between the deep temporal fascia (below) and the superficial temporal fascia (above). We believe that these vessels can be thought of as arrows pointing superiorly to a frontal branch of the facial nerve as it courses through the superficial temporal fascia. PMID- 12633202 TI - Multicenter evaluation of subcutaneous augmentation material implants. AB - OBJECTIVE: To evaluate the performance of subcutaneous augmentation material preformed shapes for facial implantation. DESIGN: One-year prospective multicenter evaluation of implant performance. SETTING: General community hospital; private and institutional practice; and ambulatory care setting. PATIENTS: Eighty-two patients undergoing elective cosmetic and reconstructive procedures. INTERVENTIONS: Patients received nasal dorsal implants (31 patients); chin implants (38 patients); and malar implants (13 patients). MAIN OUTCOME MEASURES: Complications and aesthetic outcome, including implant position, projection, contour, symmetry, and overall aesthetic index. RESULTS: Complication rates for infection that required implant removal included nasal dorsal implants, 3.2%; chin implants, 5.3%; and malar implants, 3.8%. The overall aesthetic outcome was judged by an independent panel of facial plastic surgeons on a scale from 1 (poor) to 5 (excellent): nasal dorsal implants scored 4.1; chin implants, 3.8; and malar implants, 3.6. CONCLUSIONS: This study demonstrates that subcutaneous augmentation material preformed shapes offer a relatively safe and effective treatment alternative for permanent facial augmentation. PMID- 12633203 TI - Surgical access to the internal nasal valve. AB - OBJECTIVES: To measure the width of the nasal septum in the region of the internal nasal valve using radiographic images to support use of open septorhinoplasty for alteration of the internal nasal valve and to compare short term patient satisfaction for open septorhinoplasty vs endonasal septoplasty. STUDY DESIGN: Radiographic study and chart review. METHODS: The width of the nasal septum at the level of the inferior turbinates and at the level of maximum septal thickness was measured on computed tomographic scans of 70 patients. Patient satisfaction 12 weeks after treatment was assessed in 113 patients who underwent septoplasty or open septorhinoplasty for nasal obstruction and septal and internal nasal valve abnormalities. RESULTS: Measures from the computed tomographic scans revealed that the nasal septum was significantly wider at the internal nasal valve than at the inferior aspect of the septum. All 34 patients who underwent septoplasty and all 79 patients who underwent open septorhinoplasty were satisfied with the results at 12 weeks after surgery. CONCLUSIONS: Anatomic evidence supports the need to address the superior septum. Although this may be facilitated by an open septorhinoplasty approach, short-term results showed no difference in patient satisfaction with this technique compared with septoplasty. The differences between the techniques may be evident on long-term follow-up. PMID- 12633204 TI - Total nasal reconstruction: utility of the free radial forearm fascial flap. AB - BACKGROUND: Total nasal defects present daunting challenges to the reconstructive surgeon. The nasal skeleton can be successfully fabricated with bone and cartilage. Reconstruction of the nasal skin with a forehead flap produces an excellent color match for nasal skin. Resurfacing of the internal lining is the most difficult of the 3 layers. Local tissue is often unsatisfactory in amount and/or vascular supply. METHODS: A patient requiring total nasal reconstruction was prospectively examined. Intraoperative technique was recorded, and postoperative function was determined. RESULTS: A paramedian forehead flap was used to resurface the external defect. Split calvarium and conchal cartilage were used to reconstruct the nasal skeleton. A fascial flap harvested from the forearm was used to replace the intranasal lining. Turbinate grafts were placed to line the flap. Postoperative breathing was excellent. CONCLUSIONS: The intranasal portion of a total nasal defect can be successfully reconstructed with a fascial forearm flap. Placement of a turbinate or mucosal graft allows for a thin mucosalized lining with an excellent functional outcome. PMID- 12633205 TI - Evaluation of pericranial skull adherence and periosteal healing in the rabbit model. PMID- 12633207 TI - The platinum chain: a new upper-lid implant for facial palsy. AB - BACKGROUND: When using upper-lid gold implants in facial palsy, a certain percentage of undesired effects and complications are known to occur (eg, astigmatism, pseudoptosis, migration, bulging, and extrusion). Our flexible platinum chain implant adapts better to the changing radius of the tarsus with movement of the globe and reduces the complication rate. Platinum implants are smaller, owing to their higher density. OBJECTIVE: To compare the results with 33 gold and 30 platinum chain implantations. METHODS: Clinical follow-up study at a university otorhinolaryngology department. The mean follow-up was 11 months in the gold-implant group and 9 months in the platinum-implant group. The treatment consisted of pretarsal fixation of the metal weights to the upper lid. MAIN OUTCOME MEASURES: Criteria for evaluation of results include reduction of lagophthalmos and keratopathy and gain of visual acuity. Complications that have been assessed include astigmatism, bulging, migration, pseudoptosis, and extrusion of implants. We evaluated histological samples of the implant bed and performed ultrasound measurements of the tarsal radius. RESULTS: The restoration of lid closure was a visual and aesthetic improvement for all patients. We found a tendency toward a higher rate of complications with gold implants compared with platinum chains, especially for astigmatism and bulging. Ultrasonographic measurements showed ongoing flexibility of the platinum chains after implantation, and histological findings confirmed their good biocompatibility. CONCLUSIONS: Flexible platinum chain implants lead to better results with fewer complications compared with standard rigid gold implants. PMID- 12633208 TI - Minimally invasive brow suspension for facial paralysis. AB - OBJECTIVE: To report a new technique for unilateral brow suspension for facial paralysis that is minimally invasive, limits supraciliary scar formation, does not require specialized endoscopic equipment or expertise, and has proved to be equal to direct brow suspension in durability and symmetry. DESIGN: Retrospective survey of a case series of 23 patients between January 1997 and December 2000. SETTING: Metropolitan tertiary care center. PATIENTS: Patients with head and neck tumors and brow ptosis caused by facial nerve paralysis. MAIN OUTCOME MEASURE: The results of the procedure were determined using the following 3-tier rating system: outstanding (excellent elevation and symmetry); acceptable (good elevation and fair symmetry); and unacceptable (loss of elevation). RESULTS: The results were considered outstanding in 12 patients, acceptable in 9 patients, and unacceptable in only 1 patient. One patient developed a hematoma, and 1 patient required a secondary adjustment. CONCLUSIONS: The technique has proved to be superior to standard brow suspension procedures with regard to scar formation and equal with respect to facial symmetry and suspension. These results have caused us to abandon direct brow suspension and to use this minimally invasive method in all cases of brow ptosis due to facial paralysis. PMID- 12633209 TI - Cleft lip nasal reconstruction using porous high-density polyethylene. AB - BACKGROUND: The multitude of factors involved with a unilateral cleft lip nasal defect has spurred various surgical techniques in the past. Recently, synthetic materials have been introduced for use in nasal reconstruction. OBJECTIVE: To report on and illustrate the use of porous high-density polyethylene implants in cleft lip nasal reconstruction. DESIGN: A retrospective review of cleft lip nasal reconstruction using porous high-density polyethylene in patients with a unilateral cleft lip defect from January 1, 1993, through June 30, 2000. SETTING: Facial plastic surgery private practice. PATIENTS: Eighteen patients with a unilateral cleft lip without a history of formal rhinoplasty. INTERVENTIONS: All 18 patients required multiple implants, including a columellar strut, premaxillary and prealveolar plumper grafts, a dorsal tip implant, and a unilateral nasal valve batten, using the open rhinoplasty approach. RESULTS: Favorable aesthetic results, as judged by one of us (T.R.), were achieved in all patients. All implants were well tolerated. Postoperative follow-up ranged from 6 months to 7 years. A complication occurred in 1 patient (6%), which resolved with removal of a single implant and intravenous antibiotic therapy. No other complications, including skin erosion or implant extrusion, have been noted. CONCLUSIONS: Porous high-density polyethylene implants for cleft lip nasal reconstruction are well tolerated and achieve good aesthetic results. Porous high density polyethylene implants lend stability through fibrovascular ingrowth, with integration of the implants to the surrounding tissue. PMID- 12633210 TI - Submentoplasty as an isolated rejuvenative procedure for the neck. AB - As facial cosmetic surgery has become more popular, patients' desires for minimally invasive surgery and brief healing times have increased. Submentoplasty has been described as an isolated procedure, but it is more commonly incorporated into a face-lift for rejuvenation of the cheeks, jowls, and neck. However, many patients considering submentoplasty do not need or desire face-lift surgery. Submentoplasty, in combination with liposuction, wide undermining, and platysma plications, has proven to be an effective way to rejuvenate the neck by restoring the cervicomental angle without large-incision surgery such as a face-lift. Presented here is a technique for performing submentoplasty developed over experience with more than 100 patients of widely varying ages. Recovery time is shorter than with standard face-lift techniques, and complications have been few. Submentoplasty is a viable option for patients who desire rejuvenation of the neck as a solo procedure or in combination with other facial cosmetic surgery. PMID- 12633211 TI - Isolated platysmaplasty: a useful procedure but with important limitations. PMID- 12633212 TI - Evaluation of nasoalveolar cysts for the facial plastic surgeon. AB - Asymmetrical alar flare without a history of trauma, surgery, or congenital clefting is extremely rare; therefore, a thorough investigation into the underlying reasons for the asymmetry should be undertaken before a unilateral alar base resection is performed. Nasoalveolar cyst is a readily apparent diagnosis that should not be missed after proper intranasal and intraoral examination. Excision is recommended via a sublabial approach. We describe a patient in whom a unilateral alar base reduction was initially recommended, when in fact she had a nasoalveolar cyst. We also discuss the histologic characteristics, embryological pattern, and differential diagnosis. PMID- 12633213 TI - A novel, conformable, rapidly setting nasal splint material: results of a prospective study. AB - OBJECTIVE: To determine if a fiberglass splint material (3M Scotchcast, St Paul, Minn; Smith & Nephew, Inc, Charlotte, NC) serves as a better nasal cast than Aquaplast (Smith & Nephew, Inc) based on the criteria of total preparation time, ease of use, conformability, adhesion, and patient acceptability. DESIGN: Prospective randomized comparison study with statistical analysis. SETTING: University-based division of a facial plastic surgery private clinic. PATIENTS: Sixteen consecutive patients desiring a functional and aesthetic improvement underwent open structure rhinoplasty with lateral osteotomies. No patients withdrew because of adverse effects. INTERVENTION: Half of the patients were randomized to the study group (fiberglass splint material) and half to the control group (Aquaplast). Each patient was fitted with the appropriate casting material at the conclusion of the rhinoplasty procedure. Data were collected with respect to total preparation time, ease of use, conformability, and adhesion. Each patient was asked to evaluate the cast for level of comfort after 1 week. RESULTS: There were no significant differences between study groups for adhesion (P =.18) and conformability (P =.35). Patient acceptability was good or excellent in both study groups. The fiberglass splint material was significantly easier to use (P<.001) and required less total preparation time (P<.001). The mean total preparation time for the Aquaplast and fiberglass splint materials were, respectively, 7 minutes 32 seconds and 2 minutes 37 seconds. CONCLUSIONS: Compared with the commonly used nasal splinting material Aquaplast, fiberglass splint material is not significantly more adhesive, conformable, or comfortable for the patient. Fiberglass splint material is significantly easier to use and requires less total preparation time than Aquaplast. These differences may provide advantages in the immediate postoperative period and for the long-term success of rhinoplasty surgery. PMID- 12633214 TI - The use of percutaneous sutures for graft fixation in rhinoplasty. AB - We reviewed our 10-year experience using percutaneous suturing to secure cartilage grafts in rhinoplasty. A total of 382 patients having up to 4 percutaneous sutures per surgery were analyzed. Cases using this suture technique included lower lateral onlay grafts, dorsal onlay grafts, tip grafts, and intact conchal cartilage grafts to repair nasal valve collapse. Sutures are ideally placed in a horizontal mattress fashion beginning at the skin surface, continuing into the nasal cavity, and then out through the skin. Sutures are strategically placed to secure the graft and at times stent open the nasal valve. The suture is tied loosely over the skin taking in to consideration the anticipated edema formation. There is no need to use a bolster technique. Sutures are removed at the second postoperative visit. Long-term follow-up reveals precise graft placement. There are in most instances no visible suture marks and our results achieve over 95% patient satisfaction. Percutaneous suture placement is a simple technique that allows precise graft placement and fixation without resultant scarring. It coapts the skin and soft tissues to the grafted nasal skeleton and prevents fluid accumulation, hematoma formation, and graft migration. We believe that it aids in decrease long-term tissue edema and excessive fibrosis, therefore promoting quicker healing. Our patients are both functionally and cosmetically pleased. PMID- 12633215 TI - The cervicodeltopectoral flap for single-stage resurfacing of anterolateral defects of the face and neck. AB - OBJECTIVE: To evaluate prospectively the feasibility and utility of adding a cervical extension to the standard deltopectoral flap. We postulated that this cervicodeltopectoral (CDP) flap will allow for single-stage reconstruction of large defects of the anterolateral face and neck. METHODS: As is the case with the deltopectoral flap, the CDP flap is based on the perforating branches of the internal mammary artery. However, the superior and posterior limbs of the CDP flap are not limited by the clavicle and the deltoid muscle. Instead, they extend in a subcutaneous tissue plane to the margins of the neck or facial defect and transfer the entire intervening skin bridge with the deltopectoral flap. RESULTS: Eighteen patients underwent closure of complex cutaneous defects of the face and neck with the CDP flap. We found no evidence of flap loss in any of these patients. Twelve patients had received preoperative radiation therapy encompassing the cervical extension of the CDP flap. No evidence of adverse healing was noted in this subset of patients. CONCLUSIONS: The CDP flap may represent an alternative in the surgical treatment of various cutaneous defects of the face and neck. It allows for single-stage, reliable reconstruction of these defects. The transfer of intervening cervical skin in conjunction with the deltopectoral flap provides for a more aesthetically pleasing reconstruction, as skin immediately adjacent to the defect is more closely related to the excised skin in terms of color and texture. PMID- 12633216 TI - Periosteal healing. PMID- 12633217 TI - Excision vs Mohs surgery for basal cell carcinoma. PMID- 12633220 TI - Beauty: Egon Schiele: expressive sensuality. PMID- 12633221 TI - Resurgence in quasiclassical scattering. AB - In quasiclassical spectral theory, "resurgence" means that long periodic orbits can be expressed by short ones in such a way that the spectral determinant is real. The question has thus long been posed whether long scattering orbits can be expressed by short orbits in such a way as to make the quasiclassical scattering matrix unitary. We here find a resurgent and manifestly Hermitean expression for Wigner's R matrix, implying a unitary scattering matrix. The result is particularly important if the average resonance width is comparable with the average resonance spacing. PMID- 12633222 TI - Optimal lattice domain-wall fermions. AB - I show that the conventional formulations of lattice domain-wall fermion with any finite N(s) (in the fifth dimension) do not preserve the chiral symmetry optimally and propose a new action which preserves the chiral symmetry optimally for any finite N(s). PMID- 12633225 TI - Raman regime energy dependence of alignment and orientation of KrII states populated by resonant Auger effect. AB - The energy dependencies of alignment parameters A20 for KrII 4p(4)5p states after the Auger decay of the KrI 3d(9)np resonances were investigated theoretically and experimentally for the first time in the Raman regime with the bandwidth of the exciting radiation (deltaE(FWHM)=20 meV) smaller than the natural width of the resonances (Gamma approximately 80 meV). The observed energy dependence is due to the in-terference between the different resonance channels and the direct photoionization channel. A strong energy dependence for both the orientation parameter O10 and the photoelectron angular distribution parameter beta(el) is also predicted. PMID- 12633224 TI - Measurement of the tensor analyzing powers T20 and T21 in elastic electron deuteron scattering. AB - The tensor analyzing power components T20 and T21 have been measured in elastic electron-deuteron scattering at the 2 GeV electron storage ring VEPP-3, Novosibirsk, in a four-momentum transfer range from 8.4 to 21.6 fm(-2). A new polarized internal gas target with an intense cryogenic atomic beam source was used. The new data determine the deuteron form factors G(C) and G(Q) in an important range of momentum transfer where the first node of the deuteron monopole charge form factor is located. The new results are compared with previous data and with some theoretical predictions. PMID- 12633226 TI - Transition from molecular complex to quantum solvation in 4HeNOCS. AB - We present fully quantum calculations of the rotational energy levels and spectroscopic rotational constants of the linear OCS molecule in variable size clusters of 4He. The rotational constants of OCS are found to decrease monotonically from the gas phase value as the number of helium atoms increases to N=6, after which the average constant increases to saturation at the large droplet value by N=20. The minimum is shown to indicate a transition from a molecular complex to a quantum solvated molecule, with the former characterized by floppy but near rigid behavior, while the latter is characterized by nonzero permutation exchanges and a smaller extent of rigid coupling. PMID- 12633228 TI - Attosecond spectral shearing interferometry. AB - We show that the complete characterization of arbitrarily short isolated attosecond x-ray pulses can be achieved by applying spectral shearing interferometry to photoelectron wave packets. These wave packets are coherently produced through the photoionization of atoms by two time-delayed replicas of the x-ray pulse, and are shifted in energy with respect to each other by simultaneously applying a strong laser field. The x-ray pulse is reconstructed with the algorithm developed for optical pulses, which requires no knowledge of ionization physics. Using a 800-nm shearing field, x-ray pulses shorter than approximately 400 asec can be fully characterized. PMID- 12633227 TI - Lever-assisted two-noise stochastic resonance. AB - A critical interplay of two correlated noises in a nonlinear symmetrical two-well potential system is experimentally demonstrated. One state can become completely noise free, leading to an infinite Kramers time. If an independent lever breaks the potential symmetry, stochastic resonance is recovered. In this new regime, we obtain a plateau, i.e., a high signal-to-noise ratio even for vanishing forcing signals. PMID- 12633229 TI - Emergence and control of multiphase nonlinear waves by synchronization. AB - Large amplitude multiphase solutions of the periodic Korteweg-de Vries equation are excited and controlled by a small forcing. The approach uses passage through an ensemble of resonances and subsequent multiphase self-locking of the system with eikonal-type perturbations. The synchronization of each phase in the Korteweg-de Vries wave is robust, provided the corresponding driving amplitude exceeds a threshold. PMID- 12633230 TI - Free surface waves in wall-bounded granular flows. AB - We report free-surface waves in granular flows near boundaries in an inclined chute. The chevron-shaped traveling waves spontaneously develop at inclinations close to the angle of repose for both steady and accelerating flows. Two distinct regimes are characterized by internal angle and frequency variations. Experimental measurements indicate that subsurface circulation driven by velocity gradients near frictional walls plays a central role in the pattern formation mechanism, suggesting that wave generation is controlled by the granular analog of a fluid boundary layer. PMID- 12633223 TI - Measurement of branching fractions for B-->eta(c)K(*) decays. AB - We report measurements of branching fractions for charged and neutral B-->eta(c)K decays where the eta(c) meson is reconstructed in the K(0)(S)K+/-pi(-/+), K+K pi(0), K(*0)K-pi(+), and pp; decay channels. The neutral B0 channel is a CP eigenstate and can be used to measure the CP violation parameter sin(2phi(1). We also report the first observation of the B0-->eta(c)K(*0) mode. The results are based on an analysis of 29.1 fb(-1) of data collected by the Belle detector at KEKB. PMID- 12633231 TI - Geometry and physics of wrinkling. AB - The wrinkling of thin elastic sheets occurs over a range of length scales, from the fine scale patterns in substrates on which cells crawl to the coarse wrinkles seen in clothes. Motivated by the wrinkling of a stretched elastic sheet, we deduce a general theory of wrinkling, valid far from the onset of the instability, using elementary geometry and the physics of bending and stretching. Our main result is a set of simple scaling laws; the wavelength of the wrinkles lambda approximately K(-1/4), where K is the stiffness due to an "elastic substrate" effect with a multitude of origins, and the amplitude of the wrinkle A approximately lambda. These could form the basis of a highly sensitive quantitative wrinkling assay for the mechanical characterization of thin solid membranes. PMID- 12633232 TI - Measured local heat transport in turbulent Rayleigh-Benard convection. AB - Local convective heat flux in turbulent thermal convection is obtained from simultaneous velocity and temperature measurements in an aspect-ratio-one convection cell filled with water. It is found that fluctuations of the vertical heat flux are highly intermittent and are determined primarily by the thermal plumes in the system. The experiment reveals a unique mechanism for the heat transport in turbulent convection. PMID- 12633233 TI - Measurement of the spatial coherence function of undulator radiation using a phase mask. AB - A measurement of the horizontal coherence function of 7.9 keV radiation from an undulator beam line at the Advanced Photon Source is reported. X-ray diffraction from a phase-shifting mask was used, and the coherence function was measured as a function of the width of beam-conditioning slits in the beam line. The coherence distribution is found to be best described by a Lorentzian function. PMID- 12633234 TI - Nonlinear theory of void formation in colloidal plasmas. AB - A nonlinear time-dependent model for void formation in colloidal plasmas is proposed. For experimentally relevant initial conditions, the model describes the nonlinear evolution of a zero-frequency linear instability that grows rapidly in the nonlinear regime and subsequently saturates to form a void. A number of features of the model are consistent with experimental observations under laboratory and microgravity conditions. PMID- 12633235 TI - Enhanced spatiotemporal laser-beam smoothing in gas-jet plasmas. AB - Spatiotemporal smoothing of large-scale laser intensity fluctuations is observed for a laser beam focused into underdense helium plasmas. This smoothing is found to be severely enhanced when focusing the laser beam into a helium gas jet. In contrast to other experiments with preformed plasmas, the average and the peak laser intensities are well below the threshold for ponderomotive self-focusing. The coherence characteristics of the transmitted light are measured for various electron densities, and the smoothing effect is explained by multiple scattering of laser light on self-induced density perturbations. PMID- 12633236 TI - Electron-Bernstein-wave current drive in an overdense plasma at the Wendelstein 7 AS stellarator. AB - Electron-Bernstein-wave (EBW) current drive in an overdense plasma was demonstrated at the Wendelstein 7-AS stellarator for the first time. The EBWs were generated by O-X-B mode conversion. The relatively high current drive efficiency was consistent with theoretical predictions. The experiments provided first investigations of EBW phase space interaction for wave refractive indices much larger than unity. PMID- 12633237 TI - Observation of laser-pulse-induced traveling microbubbles in dusty plasma liquids. AB - We report a direct experimental observation of traveling microbubbles induced by intense laser pulses in strongly coupled dusty plasma liquids. The dense plasma ablated from a suspended dust particle generates a spherical plasma bubble with a low dust density, in the quiescent regime before a transition to self-organized longitudinal dust density waves. It travels downwards at a velocity about 6 cm/sec inside the dust liquid. Dust density fluctuations trailing the bubble are also observed. The bubble generated in the high pressure dissipative regime collapses right after formation. PMID- 12633238 TI - Demonstration of plasma startup by coaxial helicity injection. AB - The first successful results on the transfer of a coaxial helicity injection (CHI) produced discharge to inductive operation are reported. CHI-assisted plasma startup is more robust than inductive only operation. After hand off for inductive operation, the initial 90 kA of CHI-produced current drops to 40 kA, then ramps up to 170 kA, using only 30 mV s, more than 30% higher than that produced by induction alone. These significant performance enhancing results were obtained on the HIT-II spherical torus experiment (major/minor radius of 0.3/0.2 m). PMID- 12633239 TI - Interface motion and nucleation of solid helium-4 induced by acoustic waves. AB - Crystallization and melting of helium-4 was experimentally observed to be induced by acoustic radiation pressure where the liquid-solid interface is highly mobile at low temperatures. We discuss the observed anomalous reversal of this effect as a function of temperature and the nucleation by acoustic wave pulses of crystals in the liquid phase or liquid bubbles in the solid phase. A high-speed camera was used for the in situ observation of large interface velocities as high as 1 m/sec. PMID- 12633240 TI - Thermoelastic mechanism for logarithmic slow dynamics and memory in elastic wave interactions with individual cracks. AB - Logarithmic-in-time slow dynamics has been found for individual cracks in a solid. Furthermore, this phenomenon is observed during both the crack acoustic conditioning and the subsequent relaxation. A thermoelastic mechanism is suggested which relates the log-time behavior to the essentially 2D character of the heating and cooling of the crack perimeter and inner contacts. Nonlinear perturbation of the contacts by a stronger (pump) wave causes either softening or hardening of the sample, and induces either additional absorption or transparency for a weaker (probe) acoustic wave depending on frequency of the latter. PMID- 12633241 TI - Temperature-dependent hydrogen-bond geometry in liquid water. AB - We have determined the hydrogen-bond geometry in liquid water from 0 to 80 degrees C by combining measurements of the proton magnetic shielding tensor with ab initio density functional calculations. The resulting moments of the distributions of hydrogen-bond length and angle are direct measures of thermal disorder in the hydrogen-bond network. These moments, and the distribution functions that can be reconstructed from them, impose quantitative constraints on structural models of liquid water. PMID- 12633242 TI - Predicting crystal structures: the Parrinello-Rahman method revisited. AB - By suitably adapting a recent approach [A. Laio and M. Parrinello, Proc. Natl. Acad. Sci. U.S.A. 99, 12 562 (2002)]] we develop a powerful molecular dynamics method for the study of pressure-induced structural transformations. We use the edges of the simulation cell as collective variables and define a metadynamics that drives the system away from the local minimum towards a new crystal structure. In contrast to the Parrinello-Rahman method, our approach shows no hysteresis, and crystal structure transformations can occur at the equilibrium pressure. We illustrate the power of the method by studying the pressure-induced diamond to simple hexagonal phase transition in a model of silicon. PMID- 12633243 TI - Glass breaks like metal, but at the nanometer scale. AB - We report in situ atomic force microscopy experiments which reveal the presence of nanoscale damage cavities ahead of a stress-corrosion crack tip in glass. Their presence might explain the departure from linear elasticity observed in the vicinity of a crack tip in glass. Such a ductile fracture mechanism, widely observed in the case of metallic materials at the micrometer scale, might be also at the origin of the striking similarity of the morphologies of fracture surfaces of glass and metallic alloys at different length scales. PMID- 12633244 TI - Wetting-to-nonwetting transition in metal-coated C60. AB - Based on ab initio and density-functional theory calculations, an empirical potential is proposed to model the interaction between a fullerene molecule and many sodium atoms. This model predicts homogeneous coverage of C60 below eight Na atoms and a progressive droplet formation above this size. The effects of ionization, temperature, and external electric field indicate that the various, and apparently contradictory, experimental results can indeed be put into agreement. PMID- 12633245 TI - Following adsorption kinetics at electrolyte/metal interfaces through crystal truncation scattering: sulfur on Au(111). AB - Combining electrochemical methods, in situ scanning tunneling microscopy, and surface x-ray diffraction allowed study of the structure and kinetics of S/Au(111) electrodes in aqueous electrolytes under potential control. Integrated intensities of a particular crystal truncation rod at anti-Bragg conditions were used to trace the sulfur adsorption and desorption as a function of electrode potential in real time. The S desorption is a first order process and the adsorption follows a Langmuir isotherm. A weakly bound S layer is found on the surface before charge transfer, and then specific adsorption occurs. PMID- 12633246 TI - Vibrational phase relaxation along the critical isochore of nitrogen: the role of local density fluctuations in the rate enhancement. AB - Vibrational dephasing of the nitrogen molecule is known to show highly interesting anomalies near its gas-liquid critical point. Here we present theoretical and computational studies of the Raman linewidth of nitrogen along the critical isochore. The linewidth is found to have a lambda-shaped temperature dependence near the critical point. As observed in experimental studies, the calculated line shape becomes Gaussian as the critical temperature (T(c)) is approached. Both the present simulation and a mode coupling theory analysis show that the slow decay of the enhanced density fluctuations near the critical point, probed at the subpicosecond time scales by vibrational frequency modulation, along with an enhanced vibration-rotation coupling, are the main causes of the observed anomalies. PMID- 12633247 TI - Chain-length-dependent relaxation scenarios in an oligomeric glass-forming system: from merged to well-separated alpha and beta loss peaks. AB - We have studied the relaxation dynamics of a homologous series of propylene glycol based dimethyl ethers in the supercooled regime by means of broadband dielectric spectroscopy. The system is chosen in order to minimize changes of the intermolecular interactions with varying molecular weight, M. A gradual transformation from a scenario of well-separated to one of merged alpha and beta loss peaks was observed with decreasing M. The results give strong evidence for the currently debated excess wing being due to an underlying beta relaxation. The study suggests that the main difference between glass formers with and without excess wings is the relaxation time at the merging temperature. PMID- 12633248 TI - Mechanisms of diffusion of boron impurities in SiO2. AB - We report first-principle total-energy calculations that clarify mechanisms of boron diffusion in SiO2. We find that a B atom takes a variety of stable and metastable geometries depending on its charge state. We also find that atomic rearrangements during the diffusion manifest a wealth of bonding feasibility in SiO2 and that the calculated activation energy agrees with the experimental data available. Recombination enhanced diffusion is also proposed. PMID- 12633249 TI - Non-arrhenius behavior of the island density in metal heteroepitaxy: Co on Cu(001). AB - We present a combined theoretical and experimental study of island nucleation and growth in the deposition of Co on Cu(001)-a prototype for understanding heteroepitaxial growth involving intermixing. Experimentally, ion scattering is employed. Using density-functional theory, we obtain energy barriers for the various elementary processes and incorporate these into a kinetic Monte Carlo program to simulate the heteroepitaxial growth. Both the simulations and the experiments show a unique N-shape dependence of the island density on temperature that stems from the interplay and competition of the different processes involved. PMID- 12633250 TI - Metastability of ultradense arrays of quantum dots. AB - We present a linear stability analysis of ultradense arrays of coherently strained islands against Ostwald ripening. Surprisingly, short-range elastic interactions are found to overcome the destabilizing contribution of surface energy, leading to a metastable array of quantum dots. Simulations of Ostwald ripening kinetics directly verify the existence of this metastable regime and confirm the nature of the most unstable mode for subcritical island coverage. PMID- 12633251 TI - First-principle calculations of the experimental vibrational spectrum of a surface adsorbate: anharmonic resonance coupling between fundamental and binary modes. AB - By including relevant physical properties in our modeling of a surface adsorbate system we can, from first principles, correctly calculate all experimentally observed features in the vibrational spectrum of CH3CH2O- and CD3CH2O- adsorbed on Cu(100). That is, we reproduce the number of observed modes, the vibrational frequencies, and intensities of the modes including the presence/absence of binary overtone and combination modes. No scaling was performed. Our calculations show that the anharmonic terms of the potential energy surface of free ethanol are transferrable to the corresponding surface adsorbate, ethoxy, while the harmonic terms are not. PMID- 12633252 TI - Alternating layer and island growth of Pb on Si by spontaneous quantum phase separation. AB - Real-time in situ x-ray studies of continuous Pb deposition on Si(111)-(7x7) at 180 K reveal an unusual growth behavior. A wetting layer forms first to cover the entire surface. Then islands of a fairly uniform height of about five monolayers form on top of the wetting layer and grow to fill the surface. The growth then switches to a layer-by-layer mode upon further deposition. This behavior of alternating layer and island growth can be attributed to spontaneous quantum phase separation based on a first-principles calculation of the system energy. PMID- 12633253 TI - Excited-state forces within a first-principles Green's function formalism. AB - We present a new first-principles formalism for calculating forces for optically excited electronic states using the interacting Green's function approach with the GW Bethe-Salpeter-equation method. This advance allows for efficient computation of gradients of the excited-state Born-Oppenheimer energy, allowing for the study of relaxation, molecular dynamics, and photoluminescence of excited states. The approach is tested on photoexcited carbon dioxide and ammonia molecules, and the calculations accurately describe the excitation energies and photoinduced structural deformations. PMID- 12633254 TI - Resistance noise scaling in a dilute two-dimensional hole system in GaAs. AB - We have measured the resistance noise of a two-dimensional (2D) hole system in a high mobility GaAs quantum well, around the 2D metal-insulator transition (MIT) at zero magnetic field. The normalized noise power S(R)/R(2) increases strongly when the hole density p(s) is decreased, increases slightly with temperature (T) at the largest densities, and decreases strongly with T at low p(s). The noise scales with the resistance, S(R)/R(2) approximately R2.4, as for a second order phase transition such as a percolation transition. The p(s) dependence of the conductivity is consistent with a critical behavior for such a transition, near a density p(*) which is lower than the observed MIT critical density p(c). PMID- 12633255 TI - Electron emission from metal surfaces by ultrashort pulses: determination of the carrier-envelope phase. AB - The phase varphi of the field oscillations with respect to the peak of a laser pulse influences the light field evolution as the pulse length becomes comparable to the wave cycle and, hence, affects the interaction of intense few-cycle pulses with matter. We theoretically investigate photoelectron emission induced by an intense, few-cycle laser pulse from a metal surface (jellium) within the framework of time-dependent density functional theory and find a pronounced varphi dependence of the photocurrent. Our results reveal a promising route to measuring varphi of few-cycle light pulses (tau<6 fs at lambda=0.8 microm) at moderate intensity levels (I(p) approximately 10(12) W/cm(2)) using a solid-state device. PMID- 12633256 TI - Non-fermi-liquid behavior of quasi-one-dimensional pseudogap materials. AB - We study the spectral function of the pseudogap phase of quasi-one-dimensional charge density wave materials. Using a stochastic approach and emphasizing an exact treatment of non-Gaussian order parameter fluctuations we will go beyond a usual perturbative calculation. Our results give a good fit to angle-resolved photoemission spectroscopy data and explain the absence of the Fermi edge in charge density wave materials even above the Peierls transition, indicating non Fermi-liquid behavior. PMID- 12633257 TI - Interaction-induced magnetoresistance: from the diffusive to the ballistic regime. AB - We study interaction-induced quantum correction deltasigma(alphabeta) to the conductivity tensor of electrons in two dimensions for arbitrary Ttau, where T is the temperature and tau the transport mean free time. A general formula is derived, expressing deltasigma(alphabeta) in terms of classical propagators ("ballistic diffusons"). The formalism is used to calculate the interaction contribution to the magnetoresistance in a classically strong transverse field and smooth disorder in the whole range of temperatures from the diffusive (Ttau<<1) to the ballistic (Ttau greater, similar 1) regime. PMID- 12633258 TI - Magnetoresistance of a 2D electron gas caused by electron interactions in the transition from the diffusive to the ballistic regime. AB - On a high-mobility 2D electron gas we have observed, in strong magnetic fields (omega(c)tau>1), a parabolic negative magnetoresistance caused by electron electron interactions in the regime of k(B)Ttau/ variant Planck's over 2pi approximately 1, which is the transition from the diffusive to the ballistic regime. From the temperature dependence of this magnetoresistance the interaction correction to the conductivity deltasigma(ee)(xx)(T) is obtained in the situation of a long-range fluctuation potential and strong magnetic field. The results are compared with predictions of the new theory of interaction-induced magnetoresistance. PMID- 12633259 TI - Dimensionality-dependent self-energy corrections and exchange-correlation potential in semiconductor nanostructures. AB - We study the quasiparticle gap in semiconductor nanostructures versus dimensionality and compare it to the value obtained in the local density approximation. We first develop general arguments based on the GW approach which we then substantiate numerically by a tight binding version of this theory. We show that the gap correction is dominated by a macroscopic surface self polarization term and point out its nonmonotonic behavior versus dimensionality. PMID- 12633260 TI - Nonequilibrium transport through a Kondo dot in a magnetic field: perturbation theory and poor man's scaling. AB - We consider electron transport through a quantum dot described by the Kondo model in the regime of large transport voltage V in the presence of a magnetic field B with max((V,B)>>T(K). The electric current I and the local magnetization M are found to be universal functions of V/T(K) and B/T(K), where T(K) is the equilibrium Kondo temperature. We present a generalization of the perturbative renormali-zation group to frequency dependent coupling functions, as necessitated by the structure of bare perturbation theory. We calculate I and M within a poor man's scaling approach and find excellent agreement with experiment. PMID- 12633261 TI - Current collapse in tunneling transport through benzene. AB - We investigate the electrical transport through a system of benzene coupled to metal electrodes by electron tunneling. Using electronic structure calculations, a semiquantitative model for the pi electrons of the benzene is derived that includes general two-body interactions. After exact diagonalization of the benzene model the transport is computed using perturbation theory for weak electrode-benzene coupling (golden rule approximation). We include the effect of an applied electric field on the molecular states, as well as radiative relaxation. We predict a current collapse and strong negative differential conductance due to a "blocking" state when the electrode is coupled to the para position of benzene. In contrast, for coupling to the meta-position, a series of steps in the I-V curve is found. PMID- 12633262 TI - Magnetic-field-dependent quasiparticle energy relaxation in mesoscopic wires. AB - In order to find out if magnetic impurities can mediate interactions between quasiparticles in metals, we have measured the effect of a magnetic field B on the energy distribution function f(E) of quasiparticles in two silver wires driven out of equilibrium by a bias voltage U. In a sample showing sharp distributions at B=0, no magnetic field effect is found, whereas, in the other sample, rounded distributions at low magnetic field get sharper as B is increased, with a characteristic field proportional to U. Comparison is made with recent calculations of the effect of magnetic-impurities-mediated interactions taking into account Kondo physics. PMID- 12633263 TI - Gate-controlled spin-orbit quantum interference effects in lateral transport. AB - In situ control of spin-orbit coupling in coherent transport using a clean GaAs/AlGaAs two-dimensional electron gas is realized, leading to a gate-tunable crossover from weak localization to antilocalization. The necessary theory of 2D magnetotransport in the presence of spin-orbit coupling beyond the diffusive approximation is developed and used to analyze experimental data. With this theory the Rashba contribution and linear and cubic Dresselhaus contributions to spin-orbit coupling are separately estimated, allowing the angular dependence of spin-orbit precession to be extracted at various gate voltages. PMID- 12633264 TI - Raman-active c-axis plasma modes in multilayer high-Tc cuprate superconductors. AB - The spectacular changes of the A(1g)-polarized Raman spectra of trilayer and four layer high-T(c) cuprate superconductors below T(c) observed in recent experiments can be explained in terms of a new type of excitation that occurs in these systems, a Raman active c-axis plasmon. Its frequency, intensity, resonance properties, polarization properties, and coupling to phonon modes involving vibrations of the planar oxygens are estimated and shown to agree with those of the observed A(1g) superconductivity-induced electronic peak. PMID- 12633265 TI - First-order superconducting transition near a ferromagnetic quantum critical point. AB - We address the issue of how triplet superconductivity emerges in an electronic system near a ferromagnetic quantum critical point (FQCP). Previous studies found that the superconducting transition is of second order, and T(c) is strongly reduced near the FQCP due to pair-breaking effects from thermal spin fluctuations. In contrast, we demonstrate that near the FQCP, the system avoids pair-breaking effects by undergoing a first order transition at a much larger T(c). A second order superconducting transition emerges only at some distance from the FQCP. PMID- 12633266 TI - Friedel oscillation in charge profile and position dependent screening around a superconducting vortex core. AB - We calculate microscopically the charge distribution around a vortex in type II superconductors by solving the Bogoliubov-de Gennes equation and the Poisson equation simultaneously. Our calculations show that the charge density depletion occurs in the vortex center and the Friedel oscillation appears over the coherence length when k(F)xi is small. We also calculate the density-density correlation function K(r,r(')) as a function of two spatial variables, r and r('), and find that K(r,r(')) is strongly dependent on the distance from the vortex center. We clarify the spatial dependent screening properties on the basis of the correlation function in the core region. PMID- 12633267 TI - Coexistence of antiferromagnetism and superconductivity near the quantum criticality of the heavy-fermion compound CeRhIn5. AB - We report a study on the interplay between antiferromagnetism (AFM) and superconductivity (SC) in a heavy-fermion compound CeRhIn5 under pressure P=1.75 GPa. The onset of the magnetic order is evidenced from a clear split of 115In nuclear quadrupole resonance spectrum due to the spontaneous internal field below the Neel temperature T(N)=2.5 K. Simultaneously, bulk SC below T(c)=2.0 K is demonstrated by the observation of the Meissner diamagnetism signal whose size is the same as in the exclusively superconducting phase. These results indicate that the AFM coexists homogeneously with the SC at a microscopic level. PMID- 12633268 TI - Structural and magnetic properties of Co-Fe binary alloy monolayers on W(110). AB - We present an experimental investigation of CoxFe1-x monolayers grown on flat and stepped W(110) surfaces. Atomically resolved scanning tunneling microscopy and low energy electron diffraction reveal continuous miscibility and pseudomorphic growth of Co and Fe for 0(T) before x(t) first returns at time T to its initial value x(0). For large classes of stochastic processes, we find that a scaling law of the form (T) = T(alpha)f(t/T) is obeyed. The scaling function f(s) is, to a large extent, independent of the details of the single increment distribution, while it encodes relevant statistical information on the presence and nature of temporal correlations in the process. We discuss the relevance of these results for Barkhausen noise in magnetic systems. PMID- 12633284 TI - Weakly nonlinear model for oscillatory instability in Saturn's dense rings. AB - Viscous overstability (oscillatory instability) may play an important role in the formation of small scale structure in dense planetary rings such as Saturn's B ring. We investigate the growth and saturation of such modes in local particle simulations. Starting from a hydrodynamic model, we develop a set of ordinary differential equations to model the evolution of the amplitudes of the linearly overstable modes in the nonlinear regime. The NASA/ESA space probe Cassini can make direct observations of these modes in Saturn's rings, including their sizes and temporal development. PMID- 12633285 TI - Cosmological constraints on bulk neutrinos. AB - Recent models invoking extra space-like dimensions inhabited by (bulk) neutrinos are shown to have significant cosmological effects if the size of the largest extra dimension is R greater, similar 1 fm. We consider effects on cosmic microwave background anisotropies, big bang nucleosynthesis, deuterium and 6Li photoproduction, diffuse photon backgrounds, and structure formation. The resulting constraints can be stronger than either bulk graviton overproduction constraints or laboratory constraints. PMID- 12633286 TI - Triplication of SU(5) Monopoles. AB - We investigate all spherically symmetric fundamental monopole solutions with fixed topological charge in the SU(5)-->[SU(3) x SU(2) x U(1)]/Z(3) x Z(2) symmetry breaking. We find that there are three solutions that are gauge equivalent but, as we argue, would correspond to physically distinct degrees of freedom in the dualized version of the model. The triplication of monopoles could help us understand the observed family structure of standard model particles. PMID- 12633283 TI - Search for supernova relic neutrinos at Super-Kamiokande. AB - A search for the relic neutrinos from all past core-collapse supernovae was conducted using 1496 days of data from the Super-Kamiokande detector. This analysis looked for electron-type antineutrinos that had produced a positron with an energy greater than 18 MeV. In the absence of a signal, 90% C.L. upper limits on the total flux were set for several theoretical models; these limits ranged from 20 to 130 macro nu(e) cm(-2) s(-1). Additionally, an upper bound of 1.2 macro nu(e) cm(-2) s(-1) was set for the supernova relic neutrino flux in the energy region E(nu)>19.3 MeV. PMID- 12633287 TI - Weak phase gamma using isospin analysis and time-dependent asymmetry in B(d)- >K(S)pi(+)pi(-). AB - We present a method for measuring the weak phase gamma using isospin analysis of three body B decays into Kpipi channels. Differential decay widths and time dependent asymmetry in B(d)-->K(S)pi(+)pi(-) mode needs to be measured into even isospin pipi states. The method can be used to extract gamma, as well as the size of the electroweak penguin contributions. The technique is free from assumptions like SU(3) or neglect of any contributions to the decay amplitudes. By studying different regions of the Dalitz plot, it may be possible to reduce the ambiguity in the value of gamma. PMID- 12633288 TI - Exclusive measurement of coherent proton-deuteron bremsstrahlung. AB - For the first time a high-precision proton-deuteron bremsstrahlung experiment has been performed in which all the different exit channels have been distinguished separately. High-precision cross sections and analyzing powers in one of the outgoing channels, namely, the coherent bremsstrahlung with a proton and a deuteron in the final state, are presented at 190 MeV incoming proton beam energy and are compared to calculations based on the low-energy theorem. The results of the calculations vary considerably calling for a fully microscopic calculation. However, using a recipe including the initial- and final-state interactions, the predictions come close to the data. PMID- 12633289 TI - Determination of the subthreshold state contribution in 13C(alpha,n)16O, the main neutron-source reaction for the s process. AB - The reaction rate of the stellar reaction 13C(alpha,n)16O, which is currently considered to be the main neutron source for the slow (s) process at low energies, has been rederived using the direct alpha-transfer reaction 13C(6Li,d)17O leading to the subthreshold state at 6.356 MeV in 17O. The contribution of the subthreshold state is found to be much smaller than the currently accepted predictions for the main neutron source of the s process, indicating less of a role of this reaction as the neutron source for the s process scenario in low-mass stars at the asymptotic giant branch. PMID- 12633290 TI - Observation of selectivity of coherent population transfer induced by optical interference. AB - A fs time-resolved selective control of multilevel systems using superposition of two identical, frequency-chirped fields is proposed and demonstrated. By adjusting the delay between the pulses, a selected transition of the Rb doublet was brought into the "holes" of the interference pattern and remained nonexcited, thus allowing to manipulate another transition by the laser field as if it were an isolated two-level system. Based on light interference, this technique needs neither strong driving field intensities nor controlling the chirp direction to achieve the selectivity. PMID- 12633291 TI - Ultracold collision properties of metastable alkaline-earth atoms. AB - Ultracold collisions of spin-polarized 24Mg, 40Ca, and 88Sr in the metastable 3P2 excited state are investigated based on molecular potentials obtained from ab initio calculations. We calculate the long-range interaction potentials and estimate the scattering length and the collisional loss rate as a function of magnetic field. The scattering lengths show resonance behavior due to the appearance of a molecular bound state in a purely long-range interaction potential and are positive for magnetic fields below 50 mT. A loss-rate model shows that losses should be smallest near zero magnetic field and for fields slightly larger than the resonance field, where the scattering length is also positive. PMID- 12633293 TI - Time-resolved dynamics of thermal isomerization in cesium-halide cluster anions. AB - We have performed time-resolved studies of the dynamics of thermal isomerization occurring in certain cesium-halide cluster anions. Using a pump-probe technique, we have observed the repopulation of a photodepleted isomer within an ensemble as a function of time by monitoring the photoelectron spectrum. The rates of isomerization increase and the isomer lifetimes decrease as functions of temperature. The clusters undergo a gradual phase transition from solid-like to liquid-like states with liquid-like behavior obtained at approximately 500 K, much lower than the bulk melting temperatures of approximately 900 K. PMID- 12633292 TI - Observation of collective-emission-induced cooling of atoms in an optical cavity. AB - We report the observation of collective-emission-induced, velocity-dependent light forces. One-third of a falling sample containing 3 x 10(6) cesium atoms illuminated by a horizontal standing wave is stopped by cooperatively emitting light into a vertically oriented, confocal resonator. We observe decelerations up to 1500 m/s(2) and cooling to temperatures as low as 7 microK, well below the free-space Doppler limit. The measured forces substantially exceed those predicted for a single two-level atom. PMID- 12633294 TI - Stable oscillations of a spatially chaotic wave function in a microstadium laser. AB - Laser action on a single spatially chaotic wave function is obtained as a final stable state in a nonlinear dynamical model of a stadium shaped resonant cavity with an active medium. The stable single-mode lasing state corresponds to a particular metastable resonance of the cavity which wins a competition among multiple modes with positive net linear gain and has a distinct lasing threshold. PMID- 12633295 TI - Noise activated granular dynamics. AB - We study the behavior of two particles moving in a bistable potential, colliding inelastically with each other and driven by a stochastic heat bath. The system has the tendency to clusterize, placing the particles in the same well at low drivings, and to fill all of the available space at high temperatures. We show that the hopping over the potential barrier occurs following the Arrhenius rate, where the heat bath temperature is replaced by the granular temperature. Moreover, within the clusterized "phase" one encounters two different scenarios: For moderate inelasticity, the jumps from one well to the other involve one particle at a time, whereas for strong inelasticity the two particles hop simultaneously. PMID- 12633296 TI - Proton acceleration from high-intensity laser interactions with thin foil targets. AB - Measurements of energetic proton production resulting from the interaction of high-intensity laser pulses with foil targets are described. Through the use of layered foil targets and heating of the target material we are able to distinguish three distinct populations of protons. One high energy population is associated with a proton source near the front surface of the target and is observed to be emitted with a characteristic ring structure. A source of typically lower energy, lower divergence protons originates from the rear surface of the target. Finally, a qualitatively separate source of even lower energy protons and ions is observed with a large divergence. Acceleration mechanisms for these separate sources are discussed. PMID- 12633297 TI - Superfluidity in a doped helium droplet. AB - Path-integral Monte Carlo calculations of the superfluid density throughout 4He droplets doped with linear impurities are presented. After deriving a local estimator for the superfluid density distribution, we find a decreased superfluid response in the cylindrically symmetric region of the first solvation layer. The helium in this region has a superfluid transition temperature similar to that of a two-dimensional helium system and may be responsible for previously unexplained experimental Q-branch measurements. PMID- 12633298 TI - Dynamics of fullerene coalescence. AB - Fullerene coalescence experimentally found in fullerene-embedded single-wall nanotubes under electron-beam irradiation or heat treatment is simulated by minimizing the classical action for many atom systems. The dynamical trajectory for forming a (5,5) C120 nanocapsule from two C60 fullerene molecules consists of thermal motions around potential basins and ten successive Stone-Wales-type bond rotations after the initial cage-opening process for which energy cost is about 8 eV. Dynamical paths for forming large-diameter nanocapsules with (10,0), (6,6), and (12,0) chiral indexes have more bond rotations than 25 with the transition barriers in a range of 10-12 eV. PMID- 12633299 TI - Dynamics of capillary drying in water. AB - We use atomistic simulations to address the question when capillary evaporation of water confined in a hydrocarbonlike slit is kinetically viable. Activation barriers and absolute rates of evaporation are estimated using open ensemble Monte Carlo-umbrella sampling and molecular dynamics simulations. At ambient conditions, the evaporation rate in a water film four molecular diameters thick is found to be of the order 10(5)(nm(2) s)(-1), meaning that water readily evaporates. Films more than a few nanometers thick will persist in a metastable liquid state. Dissolved atmospheric gas molecules do not significantly decrease the activation barrier. PMID- 12633300 TI - Thermoelectric power of p-doped single-wall carbon nanotubes and the role of phonon drag. AB - We measured thermoelectric power S of bulk single-wall carbon nanotube materials p doped with acids. In contrast to oxygen-exposed or degassed samples, S is very small at the lowest temperatures, increases superlinearly above a characteristic and sample-dependent T, and then levels off. We attribute this unusual behavior to 1D phonon drag, in which the depression of the Fermi energy cuts off electron phonon scattering at temperatures below a characteristic T0. This idea is supported by a model calculation in which the low temperature behavior of phonon drag is specifically related to the one-dimensional character of the electronic spectrum. PMID- 12633301 TI - Surface contribution to the anisotropy of magnetic nanoparticles. AB - We calculate the contribution of the Neel surface anisotropy to the effective anisotropy of magnetic nanoparticles of spherical shape cut out of a simple cubic lattice. The effective anisotropy arises because deviations of atomic magnetizations from collinearity and thus the energy depends on the orientation of the global magnetization. The result is second order in the Neel surface anisotropy, scales with the particle's volume, and has cubic symmetry with preferred directions [+/- 1, +/-1 , +/-1]. PMID- 12633302 TI - Metal-nonmetal transition in the boron group elements. AB - Structural competition in boron group elements has been studied by means of ab initio calculations. For boron we predict a structural change alpha-B-->alpha-Ga accompanied by a nonmetal-metal transition at a pressure of about 74 GPa. For Al and Ga we find an icosahedron based elemental modification (alpha-B) 0.22 and 0.05 eV/atom, respectively, higher in energy than the corresponding metallic ground state structures. In particular, the low energy difference for Ga raises expectations into the experimental feasibility of new modifications for these elements, especially in nanosized systems. PMID- 12633303 TI - Using metallic interlayers to stabilize abrupt, epitaxial metal-metal interfaces. AB - An approach is described for stabilizing metal-metal epitaxial interfaces using a thin metallic interlayer. Rutherford backscattering and channeling techniques along with low-energy electron diffraction and keV He+ ion backscattering are used to demonstrate that an atomically thin layer of Ti deposited at the Fe-Al interface, a system well known for considerable intermixing at room temperature, forms a thin interface alloy that prevents interdiffusion and improves epitaxial growth of Fe on Al(100). The structure is stable up to about 200 degrees C. PMID- 12633304 TI - Adsorption of thiolates to singly coordinated sites on Au111 evidenced by photoelectron diffraction. AB - The adsorption structure of methylthiolate (CH3S) adsorbed on Au(111), a long standing controversial issue, has been unambiguously determined by scanned-energy and scanned-angle S 2p photoelectron diffraction. The methylthiolate molecules are found to occupy atop sites with a S-Au distance of 2.42 +/- 0.03 A. The angular distribution of the S 2p photoelectrons due to forward scattering reveals that the S-C bond is inclined by approximately 50 degrees from the surface normal towards both the [211] and [121] (nearest-neighbor thiolate) directions. PMID- 12633305 TI - Liquid order at the interface of KDP crystals with water: evidence for icelike layers. AB - We present a surface x-ray diffraction study on the KDP-water interface in which the structure of both the crystalline and liquid part of the interface has been measured. We have been able to determine the ordering components in the liquid in both the perpendicular and parallel directions. We find interface-induced ordering in the first four layers of water molecules. The first two layers behave icelike and are strongly bound to the surface. The next two layers are more diffuse and show only minor lateral and perpendicular ordering. Subsequent layers are found to behave similar to a bulk liquid. PMID- 12633306 TI - Adsorption of Xe atoms on metal surfaces: new insights from first-principles calculations. AB - The adsorption of rare gases on metal surfaces serves as the paradigm of weak adsorption where it is typically assumed that the adsorbate occupies maximally coordinated hollow sites. Density-functional theory calculations using the full potential linearized augmented plane wave method for Xe adatoms on Mg(0001), Al(111), Ti(0001), Cu(111), Pd(111), and Pt(111), show, however, that Xe prefers low coordination on-top sites in all cases. We identify the importance of polarization and a site-dependent Pauli repulsion in actuating the site preference and the principle nature of the rare-gas atom-metal surface interaction. PMID- 12633307 TI - Direct determination of strain and composition profiles in SiGe islands by anomalous x-Ray diffraction at high momentum transfer. AB - Anomalous x-ray scattering is employed for quantitative measurements of the Ge composition profile in islands on Si(001). The anomalous effect in SiGe is enhanced exploiting the dependence of the complex atomic form factors on the momentum transfer. Comparing the intensity ratios for x-ray energies below and close to the K edge of Ge at various Bragg reflections in the grazing incidence diffraction setup, the sensitivity for the Ge profile is considerably enhanced. The method is demonstrated for SiGe dome-shaped islands grown on Si(001). It is found that the composition inside the island changes rather abruptly, whereas the lattice parameter relaxes continuously. PMID- 12633308 TI - Mechanism and dynamics of the CO-induced lifting of the Pt(100) surface reconstruction. AB - The first atomistic simulations of the CO-induced lifting of the Pt(100)-hex reconstruction have been performed. During this phase transformation the surface changes back to bulk-terminated Pt(100)-(1 x 1), whereby the surface atom density decreases by approximately 20%. The simulations reveal a mechanism collective in nature, indicating that restructuring proceeds through ejection of chains of Pt atoms. These chains explain the anisotropy as seen in scanning tunneling microscopy experiments. The restructuring rate depends nonlinearly on the CO coverage, but the absence of local clustering of CO excludes an explanation in terms of elementary reaction kinetics as proposed previously. PMID- 12633309 TI - Direct determination of the energy required to operate a single molecule switch. AB - Using a noncontact atomic-force and scanning-tunneling microscope in ultrahigh vacuum, we have measured the switching energy of a single molecule switch based on the rotation of a di-butyl-phenyl leg in a Cu-tetra-3,5 di-tertiary-butyl phenyl porphyrin. The mechanics and intramolecular conformation of the switched leg is controlled by the tip apex of the noncontact atomic-force microscope. The comparison between experimental and calculated force curves shows that the rotation of the leg requires an energy less than 100 x 10(-21) J, which is 4 orders of magnitude lower than state-of-the-art transistors. PMID- 12633310 TI - Microscopic structure of Er-related optically active centers in crystalline silicon. AB - A successful observation and analysis of the Zeeman effect on the lambda approximately 1.54 microm photoluminescence band in Er-doped crystalline MBE grown silicon are presented. The symmetry of the dominant optically active centers is conclusively established as orthorhombic I(C(2v)) with g axially approximately 18.39 and g radially approximately 0. In this way the long standing puzzle as regards the paramagnetism of optically active Er-related centers in silicon is settled. Preferential generation of a single type of an optically active Er-related center confirmed in this study is essential for photonic applications of Si:Er. PMID- 12633311 TI - Solving the ultranonlocality problem in time-dependent spin-density-functional theory. AB - It has been known for some time that the exchange-correlation potential in time dependent density-functional theory is an intrinsically nonlocal functional of the density as soon as one goes beyond the adiabatic approximation. In this paper we show that a much more severe nonlocality problem, with a completely different physical origin, plagues the exchange-correlation potentials in time-dependent spin-density functional theory. We show how the use of the spin current density as a basic variable solves this problem, and we provide an explicit local expression for the exchange-correlation fields as functionals of the spin currents. PMID- 12633312 TI - Orbital-order-induced metal-insulator transition in La(1-x)Ca(x)MnO3. AB - We present evidence that the insulator-to-metal transition in La(1-x)Ca(x)MnO3 near x approximately 0.2 is driven by the suppression of coherent Jahn-Teller distortions, originating from d-type orbital ordering. The orbital-ordered state is characterized by large long-range Q2 distortions below T(O'- O*). Above T(O'- O*) we find evidence for coexistence between an orbital-ordered and an orbital disordered state. This behavior is discussed in terms of electronic phase separation in an orbital-ordered insulating and an orbital-disordered metallic state. PMID- 12633313 TI - Dynamics of metallic stripes in cuprates. AB - We study the dynamics of metallic vertical stripes in cuprates within the three band Hubbard model based on a recently developed time-dependent Gutzwiller approximation. As doping increases, the optical conductivity shows transfer of spectral weight from the charge-transfer band towards (i) an incoherent band centered at 1.3 eV, (ii) a Drude peak, due mainly to motion along the stripe, and (iii) a low-energy collective mode which softens with doping and merges with (ii) at optimum doping in good agreement with experiment. The softening is related to the quasidegeneracy between Cu-centered and O-centered mean-field stripe solutions close to optimal doping. PMID- 12633314 TI - Phase sensitive shot noise in an Andreev interferometer. AB - We investigate nonequilibrium noise in a diffusive Andreev interferometer, in which currents emerging from two normal metal/superconductor (N-S) interfaces can interfere. We observe a modulation of the shot noise when the phase difference between the two N-S interfaces is varied by a magnetic flux. This is the signature of phase-sensitive fluctuations in the normal metal. The effective charge inferred from the shot noise measurement is close to q(eff) = 2e but shows phase-dependent deviations from 2e at finite energy, which we interpret as being due to pair correlations. Experimental data are in good agreement with predictions based on an extended Keldysh Green's function approach. PMID- 12633315 TI - Quantum dots with disorder and interactions: a solvable large-g limit. AB - We analyze the problem of interacting electrons on a ballistic quantum dot with chaotic boundary conditions, where the effective interactions at low energies are characterized by Landau parameters. When the dimensionless conductance g of the dot is large, the disordered interacting problem can be solved in a saddle-point approximation which becomes exact as g --> infinity (as in a large-N theory), leading to a phase transition in each Landau interaction channel. In the weak coupling phase constant charging and exchange interactions dominate the low energy physics, while the strong-coupling phase displays a spontaneous distortion of the Fermi surface, smeared out by disorder. PMID- 12633316 TI - Spontaneous magnetism of quantum dot lattices. AB - The magnetism of square lattices of quantum dots with up to 12 electrons per dot is studied using the spin-density functional formalism. At small values of the lattice constant, all lattices are nonmagnetic and gapless. When the lattice constant is increased, the shell structure of the single dots governs the magnetism of the lattice. At closed shells, the lattices are nonmagnetic and have a gap at the Fermi level. At the beginning and at the end of a shell, they become ferromagnetic and stay gapless up to large values of the lattice constant. Antiferromagnetism was observed only at midshell after a band gap was opened. PMID- 12633317 TI - Theory for slightly doped antiferromagnetic mott insulators. AB - New trial wave functions, constructed explicitly from the unique Mott insulating state with antiferromagnetic order, are proposed to describe the ground state of a Mott insulator slightly doped with holes or electrons. A rigid band is observed as charged quasiparticles with well-defined momenta being realized in these states. These states have much less superconducting correlations than previously studied ones. Small Fermi patches obtained are consistent with recent experiments on high T(c) cuprates doped lightly with holes or electrons. PMID- 12633318 TI - Doubled full shot noise in quantum coherent superconductor-semiconductor junctions. AB - We performed low temperature shot noise measurements in superconductor (TiN) strongly disordered normal metal (heavily doped Si) weakly transparent junctions. We show that the conductance has a maximum due to coherent multiple Andreev reflections at low energy and that the shot noise is then twice the Poisson noise (S = 4eI). When the subgap conductance reaches its minimum at finite voltage the shot noise changes to the normal value (S = 2eI) due to a large quasiparticle contribution. PMID- 12633319 TI - Intrinsic pinning of vortices as a direct probe of the nonuniform Larkin Ovchinnikov-Fulde-Ferrell state in layered superconductors. AB - Previously the search for the modulated superconducting Larkin-Ovchinnikov-Fulde Ferrell (LOFF) state was performed by means of measurements which do not give direct information on spatial modulation of the superconducting state. We propose to measure interlayer conductivity in Josephson-coupled layered superconductors as a function of the strength and the orientation of the parallel magnetic field. We show that interlayer critical current and the conductivity have peaks when the magnetic field is perpendicular to the in-plane wave vector of the LOFF state and when the period of the Josephson vortex lattice induced by the magnetic field is commensurate with the LOFF period. PMID- 12633320 TI - Current in narrow channels of anisotropic superconductors. AB - We argue that, in channels cut out of anisotropic single crystal superconductors and narrow on the scale of London penetration depth, the persistent current must cause the transverse phase difference provided the current does not point in any of the principal crystal directions. The difference is proportional to the current value and depends on the anisotropy parameter, on the current direction relative to the crystal, and on the transverse channel dimension. An idea on how to measure the transverse phase is proposed. PMID- 12633322 TI - Length scale coupling for nonlinear dynamical problems in magnetism. AB - The dynamics of real magnets is often governed by several interacting processes taking place simultaneously at different length scales. For dynamical simulations, the relevant length scales should be coupled, and the energy transfer accurately described. We show that in this case the micromagnetic theory is not always reliable. We present a coarse-graining approach applicable to nonlinear problems, which provides a unified description of all relevant length scales, allowing a smooth, seamless coupling. The simulations performed on model systems show that the coarse-graining approach achieves nearly the precision of all-atom simulations. PMID- 12633321 TI - Impurities and quantum interference in the chains of YBa2Cu3O6+x. AB - Motivated by recent experiments, we study the electronic structure near impurities in the chains of YBa2Cu3O6+x. Using a model of proximity induced chain superconductivity, we show that a resonance state in the chain density of states is induced only by a magnetic impurity. The spatial form of the resonance reflects the particle-hole nature of chain superconductivity and therefore distinguishes it from other broken symmetry phases. Because of quantum interference effects between impurities, the chains can undergo a quantum phase transition into a polarized state. PMID- 12633323 TI - Nonlocal topological order in antiferromagnetic Heisenberg chains. AB - We demonstrate the existence of nonlocal topological (string) order in half integer-spin antiferromagnetic Heisenberg chains on macroscopic scale on the basis of analytical scaling analysis and density matrix renormalization group calculations. Strong numerical evidence leads to a conjecture that chains with S = (2m-1)/2 and m (m = integers) belong to the same topological class defined by the topological angle theta/pi = 1/m that plays a role similar to the fictitious gauge field in the fractional quantum Hall effect. PMID- 12633324 TI - Inflection point in the magnetic field dependence of the ordered moment of URu2Si2 observed by neutron scattering in fields up to 17 T. AB - We have measured the magnetic field dependence of the ordered antiferromagnetic moment and the magnetic excitations in the heavy-fermion superconductor URu2Si2 for fields up to 17 T applied along the tetragonal c axis, using neutron scattering. The decrease of the magnetic intensity of the tiny moment with increasing field does not follow a simple power law, but shows a clear inflection point, indicating that the moment disappears first at the metamagnetic transition at approximately 40 T. This suggests that the moment m is connected to a hidden order parameter psi which belongs to the same irreducible representation breaking time-reversal symmetry. The magnetic excitation gap at the antiferromagnetic zone center Q = (1,0,0) increases continuously with increasing field, while that at Q = (1.4,0,0) is nearly constant. This field dependence is opposite to that of the gap extracted from specific-heat data. PMID- 12633325 TI - Stability of ferromagnetism in the Hubbard model on the kagome lattice. AB - The Hubbard model on the kagome lattice has highly degenerate ground states (the flat lowest band) in the corresponding single-electron problem and exhibits the so-called flat-band ferromagnetism in the many-electron ground states as was found by Mielke [J. Phys. A 24, L73 (1991)]]. Here we study the model obtained by adding extra hopping terms to the above model. The lowest single-electron band becomes dispersive, and there is no band gap between the lowest band and the other band. We prove that, at half filling of the lowest band, the ground states of this perturbed model remain saturated ferromagnetic if the lowest band is nearly flat. PMID- 12633326 TI - Disentangling the exchange coupling of entangled donors in the Si quantum computer architecture. AB - We develop a theory for micro-Raman scattering by single and coupled two-donor states in silicon. We find the Raman spectra to have significant dependence on the donor exchange splitting and the relative spatial positions of the two-donor sites. In particular, we establish a strong correlation between the temperature dependence of the Raman peak intensity and the interdonor exchange coupling. Micro-Raman scattering can therefore potentially become a powerful tool to measure interqubit coupling in the development of a Si quantum computer architecture. PMID- 12633327 TI - Inelastic x-ray scattering in correlated Mott insulators. AB - We calculate the inelastic light scattering from x rays, which allows the photon to transfer both energy and momentum to the strongly correlated charge excitations. We find that the charge-transfer peak and the low-energy peak both broaden and disperse through the Brillouin zone similar to what is seen in experiments in materials such as Ca2CuO2Cl2. PMID- 12633328 TI - 63Cu nuclear relaxation in the quantum critical point system CeCu5.9Au0.1. AB - 63Cu NQR measurements of the 63Cu T1 are reported for the quantum critical point system CeCu5.9Au0.1 over temperatures ranging from 0.1 up to 4.2 K. Below approximately 1 K the magnetization recovery exhibits a stable, nonexponential decay function which we believe signals the onset of 2D quantum critical fluctuations, as has been noted in the literature. We find T1(-1) is proportional to T0.75 for the region T < 1 K. The observed temperature dependence is in agreement with a phenomenological model of non-Fermi liquid behavior based on the uniform susceptibility but is inconsistent with calculations based on susceptibility peaks identified via neutron scattering experiments. PMID- 12633329 TI - Entanglement purification for quantum computation. AB - We show that thresholds for fault-tolerant quantum computation are solely determined by the quality of single-system operations if one allows for d dimensional systems with 8 < or = d < or = 32. Each system serves to store one logical qubit and additional auxiliary dimensions are used to create and purify entanglement between systems. Physical, possibly probabilistic two-system operations with error rates up to 2/3 are still tolerable to realize deterministic high-quality two-qubit gates on the logical qubits. The achievable error rate is of the same order of magnitude as of the single-system operations. We investigate possible implementations of our scheme for several physical setups. PMID- 12633330 TI - Topological protection and quantum noiseless subsystems. AB - Encoding and manipulation of quantum information by means of topological degrees of freedom provides a promising way to achieve natural fault tolerance that is built in at the physical level. We show that this topological approach to quantum information processing is a particular instance of the notion of computation in a noiseless quantum subsystem. The latter then provides the most general conceptual framework for stabilizing quantum information and for preserving quantum coherence in topological and geometric systems. PMID- 12633331 TI - Experimental implementation of an adiabatic quantum optimization algorithm. AB - We report the realization of a nuclear magnetic resonance computer with three quantum bits that simulates an adiabatic quantum optimization algorithm. Adiabatic quantum algorithms offer new insight into how quantum resources can be used to solve hard problems. This experiment uses a particularly well-suited three quantum bit molecule and was made possible by introducing a technique that encodes general instances of the given optimization problem into an easily applicable Hamiltonian. Our results indicate an optimal run time of the adiabatic algorithm that agrees well with the prediction of a simple decoherence model. PMID- 12633332 TI - Unification of small and large time scales for biological evolution: deviations from power law. AB - We develop a unified model that describes both "micro" and "macro" evolutions within a single theoretical framework. The ecosystem is described as a dynamic network; the population dynamics at each node of this network describes the "microevolution" over ecological time scales (i.e., birth, ageing, and natural death of individual organisms), while the appearance of new nodes, the slow changes of the links, and the disappearance of existing nodes accounts for the "macroevolution" over geological time scales (i.e., the origination, evolution, and extinction of species). In contrast to several earlier claims in the literature, we observe strong deviations from power law in the regime of long lifetimes. PMID- 12633333 TI - Vortex glass and vortex liquid in oscillatory media. AB - We study the disordered, multispiral solutions of two-dimensional oscillatory media for parameter values at which the single-spiral/vortex solution is fully stable. Using the complex Ginzburg-Landau (CGLE) equation, we show that these states, heretofore believed to be static, actually evolve extremely slowly. This is achieved via a reduction of the CGLE to the evolution of the sole vortex coordinates. This true defect-mediated turbulence occurs in two distinct phases, a vortex liquid characterized by normal diffusion of spirals, and a slowly relaxing, intermittent, "vortex glass." PMID- 12633334 TI - Surface dynamics of polymer films. AB - The dynamics of supported polymer films were studied by probing the surface height fluctuations as a function of lateral length scale using x-ray photon correlation spectroscopy. Measurements were performed on polystyrene (PS) films of thicknesses varying from 84 to 333 nm at temperatures above the PS glass transition temperature. Within a range of wave vectors spanning 10(-3) to 10(-2) nm(-1), good agreement is found between the measured surface dynamics and the theory of overdamped thermal capillary waves on thin films. Quantitatively, the data can be accounted for using the viscosity of bulk PS. PMID- 12633335 TI - Glassy dynamics and flow properties of soft colloidal pastes. AB - The local dynamics and the nonlinear rheology of soft colloidal pastes are shown to exhibit a remarkable universal behavior in terms of a unique microscopic time scale. This variable is associated with structural relaxation under the combined action of local frictional forces and elastic driving forces. These results establish a link between the local dynamics of pastes and their nonlinear flow behavior and provide a unified description of paste rheology. PMID- 12633336 TI - Colloid-polymer mixtures in the protein limit. AB - We computed the phase-separation behavior and effective interactions of colloid polymer mixtures in the "protein limit," where the polymer radius of gyration is much larger than the colloid radius. For ideal polymers, the critical colloidal packing fraction tends to zero, whereas for interacting polymers in a good solvent the behavior is governed by a universal binodal, implying a constant critical colloid packing fraction. In both systems the depletion interaction is not well described by effective pair potentials but requires the incorporation of many-body contributions. PMID- 12633337 TI - Steric interactions in cellular structures formed in a water/oil/surfactant/cosurfactant mixture. AB - Dynamics of dense microemulsion droplets forming ordered cellular phases as temperature increases are investigated by neutron spin echo spectroscopy (NSE). The shape fluctuations of droplets are shown using a specific contrast. Their relaxation time tau(f) is obtained by analysis of the NSE curves, which also reveals the short-range Brownian motion (diffusion constant D0) of the "caged" droplets. The thermal dependence of D0 and tau(f) supports the notion of entropically driven interdroplet steric repulsion stabilizing the cellular phases. PMID- 12633338 TI - Optimization of robustness and connectivity in complex networks. AB - Scale-free networks rely on a relatively small number of highly connected nodes to achieve a high degree of interconnectivity and robustness to random failure, but suffer from a high sensitivity to directed attack. In this paper we describe a parameterized family of networks and analyze their connectivity and sensitivity, identifying a network that has an interconnectedness closer to that of a scale-free network, a robustness to attack closer to that of an exponential network, and a resistance to failure better than that of either of those networks. PMID- 12633339 TI - Scaling in ordered and critical random boolean networks. AB - Random Boolean networks, originally invented as models of genetic regulatory networks, are simple models for a broad class of complex systems that show rich dynamical structures. From a biological perspective, the most interesting networks lie at or near a critical point in parameter space that divides "ordered" from "chaotic" attractor dynamics. We study the scaling of the average number of dynamically relevant nodes and the median number of distinct attractors in such networks. Our calculations indicate that the correct asymptotic scalings emerge only for very large systems. PMID- 12633340 TI - Comment on "Anomalously broad Raman scattering spectrum due to two-magnon excitation in hexagonal YMnO3". PMID- 12633341 TI - Classical correlations and entanglement in quantum measurements. AB - We analyze a quantum measurement where the apparatus is initially in a mixed state. We show that the amount of information gained in a measurement is not equal to the amount of entanglement between the system and the apparatus, but is instead equal to the degree of classical correlations between the two. As a consequence, we derive an uncertainty-like expression relating the information gain in the measurement and the initial mixedness of the apparatus. Final entanglement between the environment and the apparatus is also shown to be relevant for the efficiency of the measurement. PMID- 12633342 TI - When is quantum decoherence dynamics classical? AB - A direct classical analog of quantum decoherence is introduced. Similarities and differences between decoherence dynamics examined quantum mechanically and classically are exposed via a second-order perturbative treatment and via a strong decoherence theory, showing a strong dependence on the nature of the system-environment coupling. For example, for the traditionally assumed linear coupling, the classical and quantum results are shown to be in exact agreement. PMID- 12633343 TI - Off-diagonal geometric phase for mixed states. AB - We extend the off-diagonal geometric phase [Phys. Rev. Lett. 85, 3067 (2000)]] to mixed quantal states. The nodal structure of this phase in the qubit (two-level) case is compared with that of the diagonal mixed state geometric phase [Phys. Rev. Lett. 85, 2845 (2000)]]. Extension to higher dimensional Hilbert spaces is delineated. A physical scenario for the off-diagonal mixed state geometric phase in polarization-entangled two-photon interferometry is proposed. PMID- 12633344 TI - Dynamical instability in a trimeric chain of interacting bose-einstein condensates. AB - We analyze thoroughly the mean-field dynamics of a linear chain of three coupled Bose-Einstein condensates, where both the tunneling and the central-well relative depth are adjustable parameters. Owing to its nonintegrability, entailing a complex dynamics with chaos occurrence, this system is a paradigm for longer arrays whose simplicity still allows a thorough analytical study. We identify the set of dynamics fixed points, along with the associated proper modes, and establish their stability character depending on the significant parameters. As an example of the remarkable operational value of our analysis, we point out some macroscopic effects that seem viable to experiments. PMID- 12633345 TI - B0-->D(*-)a(+)(1): chirality tests and resolving an ambiguity in the CP violation parameter 2beta+gamma. AB - We point out that the decays of B mesons into a vector meson and an axial-vector meson can distinguish between left and right-handed polarized mesons, in contrast to decays into two vector mesons. Measurements in B0-->D(*-)a(+)(1) are proposed for testing factorization and the V-A structure of the b-->c current, and for resolving a discrete ambiguity in 2beta+gamma. PMID- 12633346 TI - B-tau unification and large atmospheric mixing: a case for a noncanonical seesaw mechanism. AB - We study the second and third generation masses in the context of the minimal renormalizable SO(10) theory. We show that if the seesaw takes the noncanonical (type II) form, large atmospheric neutrino mixing angle requires b-tau unification. PMID- 12633348 TI - Experiments with a 3D double optical lattice. AB - We present a setup where we trap two different cesium hyperfine ground states in two different near-resonant optical lattices with identical topographies. We demonstrate that we can change the relative spatial phase between the lattices and we measure the equilibrium temperature as a function of the relative spatial phase. This provides a topographical chart of the optical potential. We also determine the rate at which atoms are transferred between the lattices and show that the setup is a promising candidate for implementing coherent quantum state manipulation. PMID- 12633349 TI - Above threshold ionization in tightly focused, strongly relativistic laser fields. AB - The dynamics of electrons ionized from high charge states by lasers with intensity >10(20) W/cm(2) have been studied. At these intensities vxB forces drive the electrons subsequent to ionization in a trajectory nearly parallel to the laser propagation direction. This gives rise to large energy gains as the electron rides in phase with the laser field over a long distance. Monte Carlo simulations illustrate that, unlike in case of ionization in sub- and near relativistic intensity fields (<10(19) W/cm(2)), the electron dynamics in the ultrarelativistic case are strongly influenced by the longitudinal electric fields found near the focus of a tightly focused laser. PMID- 12633350 TI - Complete two-electron spectra in double photoionization: the rare gases Ar, Kr, and Xe. AB - A new spectroscopic technique, giving complete two-dimensional e(-)-e(-) coincidence spectra in single photon double photoionization, is presented. The technique resolves the states of doubly charged ions and provides spectra of the individual electrons emitted in formation of each final dication state. Complete spectra for double photoionization of Xe, Kr, and Ar at photon energies up to 51 eV have been recorded. Overall and surprisingly, the np(4) 3P, 1D, and 1S states are populated according to their statistical weights. When the evident autoionization is excluded, the supposedly favored 3P states are in fact disfavored. Detailed information on the autoionization processes is also made available. PMID- 12633351 TI - Tuning p-wave interactions in an ultracold Fermi gas of atoms. AB - We have measured a p-wave Feshbach resonance in a single-component, ultracold Fermi gas of 40K atoms. We have used this resonance to enhance the normally suppressed p-wave collision cross section to values larger than the background s wave cross section between 40K atoms in different spin states. In addition to the modification of two-body elastic processes, the resonance dramatically enhances three-body inelastic collisional loss. PMID- 12633352 TI - Recombination of three ultracold fermionic atoms. AB - Three-body recombination of identical, spin-polarized fermionic atoms in the ultracold limit is investigated using model interactions. The mechanisms for recombination are parametrized by the "scattering volume" V(p) and described in the framework of the adiabatic hyperspherical representation. We have calculated the recombination rate K3 as a function of V(p) and have found K3 proportional, variant |V(p)|(8/3) for small |V(p)|. Recombination near a two-body Feshbach resonance can thus be significant. PMID- 12633347 TI - Q2 dependence of nuclear transparency for exclusive rho0 production. AB - Exclusive coherent and incoherent electroproduction of the rho(0) meson from 1H and 14N targets has been studied at the HERMES experiment as a function of coherence length (l(c)), corresponding to the lifetime of hadronic fluctuations of the virtual photon, and squared four-momentum of the virtual photon (-Q2). The ratio of 14N to 1H cross sections per nucleon, called nuclear transparency, was found to increase (decrease) with increasing l(c) for coherent (incoherent) rho(0) electroproduction. For fixed l(c), a rise of nuclear transparency with Q2 is observed for both coherent and incoherent rho(0) production, which is in agreement with theoretical calculations of color transparency. PMID- 12633353 TI - Cluster growing process and a sequence of magic numbers. AB - We present a new theoretical framework for modeling the cluster growing process. Starting from the initial tetrahedral cluster configuration, adding new atoms to the system, and absorbing its energy at each step, we find cluster growing paths up to the cluster sizes of more than 100 atoms. We demonstrate that in this way all known global minimum structures of the Lennard-Jones (LJ) clusters can be found. Our method provides an efficient tool for the calculation and analysis of atomic cluster structure. With its use we justify the magic number sequence for the clusters of noble gas atoms and compare it with experimental observations. PMID- 12633354 TI - Temporal shaping of femtosecond solitary pulses in photoionized media. AB - Femtosecond light pulses interacting with solids are studied. With a power close to the self-focusing threshold, an optical pulse evolves like a solitary wave with a temporal duration shrunk to a few femtoseconds through the defocusing action of multiphoton ionization. Self-steepening and space-time focusing are shown to form shock profiles, which do not prevent the pulse from keeping a shorter duration over one Rayleigh length. Theoretical estimates justify the key mechanisms leading to pulse compression, whose influence on the power spectra is finally discussed. PMID- 12633355 TI - Band-gap structure of waveguide arrays and excitation of Floquet-Bloch solitons. AB - Band-gap structure of periodic waveguide arrays is investigated in the linear and the nonlinear regimes. Excitation of array modes belonging to high bands is demonstrated. Floquet-Bloch solitons are demonstrated experimentally and shown to be a generalization of discrete solitons. PMID- 12633356 TI - New type of polariton in a piezoelectric superlattice. AB - We studied the propagation of an electromagnetic (EM) wave in a piezoelectric superlattice. Because of the piezoelectric effect, a transverse polarization can be induced by a longitudinal wave which couples strongly to the EM wave in some particular frequency regions, resulting in the creation of a new type of polariton that does not exist in ionic crystals. The forbidden band associated with the polariton is not due to the Bragg reflection, but rather to the coupling. PMID- 12633357 TI - Observation of vector vortex lattices in polarization states of an isotropic microcavity laser. AB - We experimentally investigate the formation of a vector polarization pattern from an isotropic microcavity laser. It is found that the orthogonal components of the observed pattern are localized on the geometrical rays. The connection between eigenfunctions and geometrical rays is analytically constructed by using the SU(2) coherent states. With the analytical function form, the observed vector pattern is completely reconstructed and the vector vortex lattice is apparent. PMID- 12633358 TI - Signatures of quantum stability in a classically chaotic system. AB - We experimentally and numerically investigate the quantum accelerator mode dynamics of an atom optical realization of the quantum delta-kicked accelerator, whose classical dynamics are chaotic. Using a Ramsey-type experiment, we observe interference, demonstrating that quantum accelerator modes are formed coherently. We construct a link between the behavior of the evolution's fidelity and the phase space structure of a recently proposed pseudoclassical map, and thus account for the observed interference visibilities. PMID- 12633359 TI - Coherent regimes of globally coupled dynamical systems. AB - This Letter presents a method by which the mean field dynamics of a population of dynamical systems with parameter diversity and global coupling can be described in terms of a few macroscopic degrees of freedom. The method applies to populations of any size and functional form in the region of coherence. It requires linear variation or a narrow distribution for the dispersed parameter. Although an approximation, the method allows us to quantitatively study the transitions among the collective regimes as bifurcations of the effective macroscopic degrees of freedom. To illustrate, the phenomenon of oscillator death and the route to full locking are examined for chaotic oscillators with time scale mismatch. PMID- 12633360 TI - Parametrically driven dark solitons. AB - We show that unlike the bright solitons, the parametrically driven kinks are immune from instabilities for all dampings and forcing amplitudes; they can also form stable bound states. In the undamped case, the two types of stable kinks and their complexes can travel with nonzero velocities. PMID- 12633361 TI - Shadowing high-dimensional hamiltonian systems: the gravitational N-body problem. AB - A shadow is an exact solution to a chaotic system of equations that remains close to a numerically computed solution for a long time. Using a variable-order, variable-time-step integrator, we numerically compute solutions to a gravitational N-body problem in which many particles move and interact in a fixed potential. We then search for shadows of these solutions with the longest possible duration. We find that in "softened" potentials, shadow durations are sufficiently long for significant evolution to occur. However, in unsoftened potentials, shadow durations are typically very short. PMID- 12633362 TI - Reconnection of colliding vortex rings. AB - We investigate numerically the Navier-Stokes dynamics of reconnecting vortex rings at small Reynolds number for a variety of configurations. We find that reconnections are dissipative due to the smoothing of vorticity gradients at reconnection kinks and to the formation of secondary structures of stretched antiparallel vorticity which transfer kinetic energy to small scales where it is subsequently dissipated efficiently. In addition, the relaxation of the reconnection kinks excites Kelvin waves which due to strong damping are of low wave number and affect directly only large scale properties of the flow. PMID- 12633363 TI - Barriers for transport in turbulent plasmas. AB - The control of transport due to electrostatic turbulence is investigated using test-particle simulations. We show that a barrier for the transport, that is, a region where transport is reduced, can be generated through the randomization of phases of the turbulent field. This corresponds to the annihilation of coherent structures which are present at all scales, without actually suppressing turbulence. When the barrier is active, a flux of particles towards the center of the simulation box is present inside the region where the barrier is located. PMID- 12633364 TI - Backscattering of an intense laser beam by an electron. AB - We present a novel, simple asymptotic expansion for the spectrum of radiation that is backscattered from a laser by a counterpropagating (or copropagating) electron. The solutions are presented in such a way that they explicitly show the relative merit of using an intense laser and of an energetic electron beam in x ray production in the single particle regime. Simple scaling laws are given. PMID- 12633365 TI - Decharging of complex plasmas: first kinetic observations. AB - The first experiment on the decharging of a complex plasma in microgravity conditions was conducted. After switching off the rf power, in the afterglow plasma, ions and electrons rapidly recombine and leave a cloud of charged microparticles. Because of microgravity, the particles remain suspended in the experimental chamber for a sufficiently long time, allowing precise measurements of the rest particle charge. A simple theoretical model for the decharging is proposed which agrees quite well with the experiment results and predicts the rest charge at lower gas pressures. PMID- 12633366 TI - Dynamics of a supersonic plume moving along a magnetized plasma. AB - A particle-in-cell code is used to investigate the evolution of a density plume moving through a background plasma with supersonic speed directed along the confinement magnetic field. For scale lengths representative of laboratory and auroral phenomena, the major nonlinear effects identified by the present simulations are the formation of a bipolar current system from the ballistic electrons, the appearance of transient potential layers, and the carving of deep density cavities. A 3D magnetic topology is generated by the self-consistent ballistic and diamagnetic currents that accompany highly localized potential layers. PMID- 12633367 TI - Light transport through the band-edge states of Fibonacci quasicrystals. AB - The propagation of light in nonperiodic quasicrystals is studied by ultrashort pulse interferometry. Samples consist of multilayer dielectric structures of the Fibonacci type and are realized from porous silicon. We observe mode beating and strong pulse stretching in the light transport through these systems, and a strongly suppressed group velocity for frequencies close to a Fibonacci band gap. A theoretical description based on transfer matrix theory allows us to interpret the results in terms of Fibonacci band-edge resonances. PMID- 12633368 TI - Hydrogenated silicon fullerenes: effects of H on the stability of metal encapsulated silicon clusters. AB - Ab initio calculations of H interaction on Si12M, Si18M2 (M=Cr, Mo, and W), and Zr@Si(16) fullerene (f) show relatively weak binding of H in agreement with experimental results of H free Si12M and Si18M2 clusters. Adsorption of H enhances sp(3) bonding between the Si atoms, weakens the M-Si cage interactions, and leads to distortions in the cages. Si12CrH12 has 4 mu(B) magnetic moment in contrast to zero for Si12Cr. Removal of the M atom leads to stable empty cages of Si12H12, f-Si16H16, and f-Si20H20 with large highest occupied-lowest unoccupied molecular orbital gaps of 2.5-3.0 eV, making them attractive for optoelectronic applications. PMID- 12633369 TI - Scaling relations for implantation of size-selected Au, Ag, and Si clusters into graphite. AB - The deposition of size-selected clusters represents a new route to the fabrication of truly nanometer-scale surface architectures, e.g., nanopores. We report a systematic experimental study, coupled with molecular dynamics simulations, of the implantation depths of size-selected Au7, Ag7, and Si7 clusters in the model graphite substrate. For impact energies between 1.0 and 5.5 keV, we find that the implantation depth scales linearly with the momentum of the clusters for all three types of cluster. This "universal" behavior is consistent with a (viscous) retarding force proportional to the velocity of the cluster, akin to Stokes's law. PMID- 12633370 TI - Molecular-dynamics simulations of carbon nanotubes as gigahertz oscillators. AB - Recently, Zheng and Jiang [Phys. Rev. Lett. 88, 045503 (2002)]] have proposed that multiwalled carbon nanotubes could be the basis for a new generation of nano oscillators in the several gigahertz range. In this Letter, we present the first molecular dynamics simulation for these systems. Different nanotube types were considered in order to verify the reliability of such devices as gigahertz oscillators. Our results show that these nano-oscillators are dynamically stable when the radii difference values between inner and outer tubes are of approximately 3.4 A. Frequencies as large as 38 GHz were observed, and the calculated force values are in good agreement with recent experimental investigations. PMID- 12633371 TI - Mechanisms of radiation-induced viscous flow: role of point defects. AB - Mechanisms of radiation-induced flow in amorphous solids have been investigated using molecular dynamics computer simulations. It is shown for a model glass system, CuTi, that the radiation-induced flow is independent of recoil energy between 100 eV and 10 keV when compared on the basis of defect production and that there is a threshold energy for flow of approximately 10 eV. Injection of interstitial- and vacancylike defects induces the same amount of flow as the recoil events, indicating that point-defect-like entities mediate the flow process, even at 10 K. Comparisons of these results with experiments and thermal spike models are made. PMID- 12633372 TI - Direct imaging of Sc2@C84 molecules encapsulated inside single-wall carbon nanotubes by high resolution electron microscopy with atomic sensitivity. AB - Intramolecular structure of the scandium dimetallofullerene (Sc(2)@C(84)) has been clearly revealed by high resolution transmission electron microscopy with the single-atom sensitivity. Direct observation of two Sc atoms inside each fullerene molecule has led to a successful determination of the molecular symmetry among the three possible structural isomers for the Sc(2)@C(84). The present work introduces a new electron microscopic approach to investigate individual molecular structures and demonstrates the possibility for determining the molecular isomer on a single-molecular basis. PMID- 12633373 TI - Photorefractivity in nanostructured tin-silicate glass ceramics: a radiation induced nanocluster size effect. AB - The possibility of obtaining permanent photoinduced refractive index changes, up to -10(-3), in nanostructured silica-based composites has been demonstrated in SiO(2):SnO(2) optical-grade glass ceramics exposed to ultraviolet radiation. Optical and electron paramagnetic resonance data suggest that the negative refractive index change involves a modification at the surface of the SnO2 nanoclusters, leading to a reduction of their crystalline size. PMID- 12633374 TI - Infrared-phonon-polariton resonance of the nonlinear susceptibility in GaAs. AB - Nonlinear probing of the fundamental lattice vibration of polar crystals is shown to reveal insight into higher-order cohesive lattice forces. With a free-electron laser tunable in the far infrared we experimentally investigate the dispersion of the second-order susceptibility due to the phonon resonance in GaAs. We observe a strong resonance enhancement of second harmonic light generation at half the optical phonon frequency, and in addition a minimum at a higher frequency below the phonon frequency. Measuring this frequency and comparison to a theoretical model allows the determination of competing higher-order lattice forces. PMID- 12633375 TI - C60 molecular bearings. AB - An ultralubricated system is reported which confines a C60 monolayer between graphite plates. C60 molecules act as molecular bearings, assisted by the nanogears of six-membered carbon rings between C60 molecules and graphite, in which the mean dynamical frictional forces are zero up to a high load of 100 nanonewtons. A stick-slip rolling model with a step rotation of a C60 molecule is proposed. This ultralubricated system, very promising for the realization of nano and micromachines, is expected to open a new field of molecular bearings. PMID- 12633376 TI - Dispersive phonon linewidths: the E2 phonons of ZnO. AB - Phonon linewidths can exhibit a large variation when either pressure or isotopic masses are changed. These effects yield detailed information about the mechanisms responsible for linewidths and lifetimes, e.g., anharmonicity or isotopic disorder. We report Raman measurements of the linewidth of the upper E2 phonons of ZnO crystals with several isotopic compositions and their dependence on pressure. Changes by a factor of 12 are observed at a given temperature. Comparison with calculated densities of one-phonon states, responsible for isotope scattering, and of two-phonon states, responsible for anharmonic decay, yields a consistent picture of these phenomena. Isotopic disorder broadening by 7 cm(-1) is found in samples with mixed 16O-18O content, whereas the anharmonic processes involve decay into sums and differences of two phonons. PMID- 12633377 TI - Relation between thermodynamics and kinetics of glass-forming liquids. AB - Vitrification of a supercooled liquid is often characterized by the hypothetical kinetic instability point, the Vogel-Fulcher temperature T0, and the thermodynamic one, the Kauzmann temperature T(K). The widely believed relation T0 congruent with T(K) is regarded as the supporting evidence of a direct connection between the thermodynamics and kinetics of glass-forming liquids. Here we demonstrate that T(K)/T(0) systematically increases from unity with a decrease in the fragility, contrary to the common belief. This systematic deviation may be explained by a synergistic effect between the weaker cooperativity and the stronger tendency of short-range ordering in stronger glass formers. PMID- 12633378 TI - Adatom kinetics on and below the surface: the existence of a new diffusion channel. AB - Employing density-functional theory in combination with scanning tunneling microscopy, we demonstrate that a thin metallic film on a semiconductor surface may open an efficient and hitherto not expected diffusion channel for lateral adatom transport: adatoms may prefer diffusion within this metallic layer rather than on top of the surface. Based on this concept, we interpret recent experiments: We explain why and when In acts as a surfactant on GaN surfaces, why Ga acts as an autosurfactant, and how this mechanism can be used to optimize group-III nitride growth. PMID- 12633379 TI - Potential-induced strain relaxation in au mono- and bilayer films on Pt(111) electrode surfaces. AB - In situ scanning tunneling microscopy observations of 1-2 monolayer thick Au films on Pt(111) electrodes are presented, which show a complex, potential dependent structural phase behavior of the Au film. Starting from a pseudomorphic structure at <0.35 V(Ag/AgCl), a sequence of transitions into dislocation network structures occurs with increasing potential: (1x1)--> "striped phase" --> "hex phase" in the Au bilayer, (1x1)--> striped phase in the Au monolayer. This is explained by a reduction of the in-plane stress in the Au surface layer(s) due to anion adsorption and a strain energy minimization as described by the Frenkel Kontorova model. PMID- 12633380 TI - Irreversibility and polymer adsorption. AB - Physisorption or chemisorption from dilute polymer solutions often entails irreversible polymer-surface bonding. We present a theory of the resultant nonequilibrium layers. While the density profile and loop distribution are the same as for equilibrium layers, the final layer comprises a tightly bound inner part plus an outer part whose chains make only fN surface contacts where N is chain length. The contact fractions f follow a broad distribution, P(f) approximately f(-4/5), in rather close agreement with strong physisorption experiments [H. M. Schneider, Langmuir 12, 994 (1996)]]. PMID- 12633381 TI - New (sqrt3 xsqrt 3)R30 degrees phase of Pb on Ge(111) and its consequence for the melting transition. AB - Depending on the preparation method, we find two different structures of the Pb/Ge(111) system at a nominal coverage of 4 / 3 monolayer that exhibit different melting points. One is the well studied beta phase that melts at 270 degrees C, but the other is a new and metastable phase that melts at 330 degrees C. Using surface x-ray diffraction the atomic structure of both phases is found to be surprisingly similar. The difference in melting points can be explained by the distribution of the excess Pb present on the surface, which has a direct effect on the vacancy density. We propose a modified phase diagram, in which the melting temperature of the beta phase depends strongly on coverage. PMID- 12633382 TI - Linear response calculations of lattice dynamics in strongly correlated systems. AB - We introduce a new linear response method to study the lattice dynamics of materials with strong correlations. It is based on a combination of dynamical mean field theory of strongly correlated electrons and the local density functional theory of electronic structure. We apply the method to study the phonon dispersions of Mott insulators NiO and MnO in their paramagnetic insulating state not accessible by local density functionals. Our results are in good agreement with experiment. PMID- 12633383 TI - Anomalous temperature dependence of the dephasing time in mesoscopic kondo wires. AB - We present measurements of the magnetoconductance of long and narrow quasi-one dimensional gold wires containing magnetic iron impurities in a temperature range extending from 15 mK to 4.2 K. The dephasing rate extracted from the weak antilocalization shows a pronounced plateau in a temperature region of 300-800 mK, associated with the phase breaking due to the Kondo effect. Below the Kondo temperature, the dephasing rate decreases linearly with temperature, in contradiction with standard Fermi-liquid theory. Our data suggest that the formation of a spin glass due to the interactions between the magnetic moments is responsible for the observed anomalous temperature dependence. PMID- 12633384 TI - Quantum ratchet effect for vortices. AB - We have measured a quantum ratchet effect for vortices moving in a quasi-one dimensional Josephson junction array. In this solid-state device the shape of the vortex potential energy, and consequently the band structure, can be accurately designed. This band structure determines the presence or absence of the quantum ratchet effect. In particular, asymmetric structures possessing only one band below the barrier do not exhibit current rectification at low temperatures and bias currents. The quantum nature of transport is also revealed in a universal/nonuniversal power-law dependence of the measured voltage-current characteristics for samples without/with rectification. PMID- 12633385 TI - Observation of spin-polarized surface states on ultrathin bct Mn(001) films by spin-polarized scanning tunneling spectroscopy. AB - We report the observation of a magnetic contrast of up to 20% in the scanning tunneling spectroscopy dI/dV maps obtained with Fe-coated tips on Mn(001) layers grown on an Fe(001) whisker at 370 K. These nanometer resolution microscopy results show that the layers couple antiferromagnetically. By normalizing the dI/dV curves by tunneling probability functions, we found a spin-dependent peak on the body-centered-tetragonal (bct) Mn(001) surface at +0.8 V, whose high spin polarization gives rise to the dI/dV map contrast. Band structure calculations allow one to identify the +0.8 V peak as due to two spin-polarized d(z(2)) surface states. PMID- 12633386 TI - Real-space observation of drift States in a two-dimensional electron system at high magnetic fields. AB - The local density of states of the adsorbate-induced two-dimensional electron system is studied in magnetic fields up to B=6 T. Landau quantization is observed and drift states with a width of about the magnetic length are found in agreement with theoretical predictions. At the tails of the Landau levels the states form closed paths indicating localization. These states show the expected energy dependence. A multifractal analysis applied to the data results in a nice parabolic shape of the characteristic f(alpha) spectra, but we find only a slight displacement of the origin from alpha=2.0 for the states in the center of the Landau level. PMID- 12633387 TI - Spin susceptibility of an ultra-low-density two-dimensional electron system. AB - We determine the spin susceptibility in a two-dimensional electron system in GaAs/AlGaAs over a wide range of low densities from 2x10(9) cm(-2) to 4x10(10) cm(-2). Our data can be fitted to an equation that describes the density dependence as well as the polarization dependence of the spin susceptibility. It can account for the anomalous g factors reported recently in GaAs electron and hole systems. The paramagnetic spin susceptibility increases with decreasing density as expected from theoretical calculations. PMID- 12633388 TI - Resistivity of dilute 2D electrons in an undoped GaAs heterostructure. AB - We report resistivity measurements from 0.03 to 10 K in a dilute high mobility 2D electron system. Using an undoped GaAs/AlGaAs heterojunction in a gated field effect transistor geometry, a wide range of densities, 0.16 x 10(10) to 7.5 x 10(10) cm(-2), are explored. For high densities, the results are quantitatively shown to be due to scattering by acoustic phonons and impurities. In an intermediate range of densities, a peak in the resistivity is observed for temperatures below 1 K. This nonmonotonic resistivity can be understood by considering the known scattering mechanisms of phonons, bulk, and interface ionized impurities. Still lower densities appear insulating to the lowest temperature measured. PMID- 12633389 TI - Low-temperature specific heat of the heavy-fermion superconductor PrOs4Sb12. AB - We report the magnetic field dependence of the specific-heat C of single crystals of the first Pr-based heavy-fermion superconductor Pr(Os4Sb12. The variation of C at low temperature and the magnetic phase diagram inferred from C, the resistivity and magnetization provide compelling evidence of a doublet ground state. Two distinct superconducting anomalies in C indicate an unconventional superconducting state, where the splitting may arise from a weak lifting of the ground state degeneracy. In combination this identifies Pr(Os4Sb12 as a strong contender for quadrupolar pairing, i.e., superconductivity that is neither electron-phonon nor magnetically mediated. PMID- 12633390 TI - Berry phase in magnetic superconductors. AB - In magnetic systems, electronic bands often acquire nontrivial topological structure characterized by gauge flux distribution in momentum k space. It sometimes follows that the phase of the wave functions cannot be defined uniquely over the whole Brillouin zone. In this Letter, we develop a theory of superconductivity in the presence of this gauge flux both in two- and three dimensional systems. It is found that the superconducting gap has "nodes" as a function of k where the Fermi surface is penetrated by a gauge string. PMID- 12633391 TI - Heavy quasiparticles in the ferromagnetic superconductor ZrZn2. AB - We report a study of the de Haas-van Alphen effect in the normal state of the ferromagnetic superconductor ZrZn2. Our results are generally consistent with a linear muffin-tin orbital band structure which predicts four exchange-split Fermi surface sheets. Quasiparticle effective masses are enhanced by a factor of 4.9 implying a strong coupling to magnetic excitations or phonons. ZrZn2 is unique among metallic ferromagnets in that it has a very large density of states in the ferromagnetic phase. PMID- 12633392 TI - Magnetic-field dependence of the maximum supercurrent of La2-xCexCuO4-y interferometers: evidence for a predominant dx2-y2 superconducting order parameter. AB - We performed a phase-sensitive test of the symmetry of the superconducting order parameter of the electron doped cuprate La(2-x)Ce(x)CuO(4-y) using a superconducting quantum interferometer with spatially distributed Josephson junctions. The studies were made on a thin film grown on a SrTiO3 tetracrystal substrate. The superconducting transition temperature was about 29 K which indicates that the sample is close to optimal doping. The magnetic field dependence of the critical current gives strong evidence for a predominant dx(2)( y(2)) order parameter symmetry of the sample measured. It also gives upper limits for the s-wave component in a mixed order parameter of the type s+idx(2)(-y(2)). PMID- 12633393 TI - Magnetic ground state of pure and doped CeFe2. AB - A combination of neutron elastic and inelastic, resonant x-ray scattering, and 57Fe Mossbauer experiments are used to determine the unusual magnetic ground state of CeFe2. The complementarities between different time-scale techniques may allow one to understand the dynamic features of the ground state in CeFe2 and its pseudobinary compounds, and how the frustration of Fe tetrahedra leads the appearance of antiferromagnetic fluctuations in the presence of ferrimagnetism. The resulting model can be used to rationalize many of the unusual and conflicting experimental results reported for this material in the literature. PMID- 12633394 TI - Optical properties of gold nanorings. AB - The optical response of ring-shaped gold nanoparticles prepared by colloidal lithography is investigated. Compared to solid gold particles of similar size, nanorings exhibit a redshifted localized surface plasmon that can be tuned over an extended wavelength range by varying the ratio of the ring thickness to its radius. The measured wavelength variation is well reproduced by numerical calculations and interpreted as originating from coupling of dipolar modes at the inner and outer surfaces of the nanorings. The electric field associated with these plasmons exhibits uniform enhancement and polarization in the ring cavity, suggesting applications in near-infrared surface-enhanced spectroscopy and sensing. PMID- 12633396 TI - Theoretical study of the optical manipulation of semiconductor nanoparticles under an excitonic resonance condition. AB - We theoretically study the mechanical interaction between radiation and a semiconductor nanoparticle, based on a microscopic model of confined excitons. The exerted force is calculated by using the Maxwell stress tensor expressed in terms of the microscopic response field. The numerical demonstrations clarify the following: (1) The enhancement of the force by using electronic resonance is remarkable for a particle with a radius of less than 100 nm for semiconductor materials. (2) The spectral peak position of the exerted force is considerably sensitive to nanoscale-size changes which would be useful for highly accurate size-selective manipulation. PMID- 12633395 TI - Spin-dependent electron-hole capture kinetics in luminescent conjugated polymers. AB - The recombination of electron-hole pairs injected in extended conjugated systems is modeled as a multistep interconversion relaxation in monoexcited electronic state space, mediated by electron-phonon coupling. The computed ratio of triplet to-singlet exciton formation times r=tau(T)/tau(S) increases from 0.9 for a model dimer to 2.5 for a 32-unit chain, in good agreement with recent experiments. We rationalize the conjugation-length dependence of r in terms of spin-specific energetics and mutual vibronic coupling of the excited states. PMID- 12633397 TI - Ultrafast carrier dynamics in single-wall carbon nanotubes. AB - Time-resolved carrier dynamics in single-wall carbon nanotubes is investigated by means of two-color pump-probe experiments. The recombination dynamics is monitored by probing the transient photobleaching observed on the interband transitions of the semiconducting tubes. This dynamics takes place on a 1 ps time scale which is 1 order of magnitude slower than in graphite. Transient photoinduced absorption is observed for nonresonant probing and is interpreted as a global redshift of the pi-plasmon resonance. We show that the opening of the band gap in semiconducting carbon nanotubes determines the nonlinear response dynamics over the whole visible and near-infrared spectrum. PMID- 12633398 TI - Optimal remote state preparation. AB - We prove that it is possible to remotely prepare an ensemble of noncommuting mixed states using communication equal to the Holevo information for this ensemble. This remote preparation scheme may be used to convert between different ensembles of mixed states in an asymptotically lossless way, analogous to concentration and dilution for entanglement. PMID- 12633399 TI - Secure quantum key distribution with an uncharacterized source. AB - We prove the security of the Bennett-Brassard (BB84) quantum key distribution protocol for an arbitrary source whose averaged states are basis independent, a condition that is automatically satisfied if the source is suitably designed. The proof is based on the observation that, to an adversary, the key extraction process is equivalent to a measurement in the sigma(x) basis performed on a pure sigma(z)-basis eigenstate. The dependence of the achievable key length on the bit error rate is the same as that established by Shor and Preskill [Phys. Rev. Lett. 85, 441 (2000)]] for a perfect source, indicating that the defects in the source are efficiently detected by the protocol. PMID- 12633400 TI - A wave traveling over a Hopf instability shapes the cochlear tuning curve. AB - The tuning curve of the cochlea measures how intense an input is required to elicit a given output level as a function of the frequency. It is a fundamental object of auditory theory, for it summarizes how to identify sounds on the basis of the cochlear output. A simple model is presented showing that only two elements are sufficient for establishing the cochlear tuning curve: a broadly tuned traveling wave, moving unidirectionally from high to low frequencies, and a set of mechanosensors poised at the threshold of an oscillatory (Hopf) instability. These two components generate the various frequency-response regimes needed for a cochlear tuning curve with a high slope. PMID- 12633401 TI - New interpretation of local dynamics of poly(dimethyl siloxane) observed by quasielastic neutron scattering. AB - Synthetic and natural polymers have complex dynamic behavior with distinct motions taking place on a wide range of time and length scales. For poly(dimethyl siloxane) we show that, at temperatures above the melting point, the reorientation of the CH3 groups provides a non-negligible contribution to the incoherent dynamic structure factor. Analysis of the quasielastic neutron scattering data is carried out using a model function that includes fast rotational motion of the CH3 groups and local conformational transitions between isomeric states. By using this model, detailed comparison between experimental data and theoretical predictions at distances where deviations from the traditional Rouse model are expected becomes possible. PMID- 12633402 TI - Direct observation of the transition from free to constrained single-segment motion in entangled polymer melts. AB - We report a direct determination of the time dependent mean-squared segment displacement of a polymer chain in the melt covering the transition from free to constraint Rouse relaxation along the virtual tube of the reptation model. This has been achieved by a neutron spin-echo (NSE) measurement of the segmental self correlation function as conveyed by the spin-incoherent scattering from two fully protonated polymer melts, polyethylene and polyethylene propylene. Within the scenario of de Gennes reptation model a transition of the time dependence of segmental mean-squared displacements from proportional, variant t(1/2) to proportional, variant t(1/4) is expected and clearly corroborated by the incoherent NSE results. PMID- 12633405 TI - Reevaluation of the crossover from elastic to capillary surface waves on soft gels. PMID- 12633404 TI - Scale-free networks are ultrasmall. AB - We study the diameter, or the mean distance between sites, in a scale-free network, having N sites and degree distribution p(k) proportional, variant k( lambda), i.e., the probability of having k links outgoing from a site. In contrast to the diameter of regular random networks or small-world networks, which is known to be d approximately ln(N, we show, using analytical arguments, that scale-free networks with 23, d approximately ln(N. We also show that, for any lambda>2, one can construct a deterministic scale-free network with d approximately ln(ln(N, which is the lowest possible diameter. PMID- 12633403 TI - Stochastic resonance in geomagnetic polarity reversals. AB - Among noise-induced cooperative phenomena a peculiar relevance is played by stochastic resonance. In this paper we offer evidence that geomagnetic polarity reversals may be due to a stochastic resonance process. In detail, analyzing the distribution function P(tau) of polarity residence times (chrons), we found the evidence of a stochastic synchronization process, i.e., a series of peaks in the P(tau) at T(n) approximately (2n+1)T(Omega)/2 with n=0,1,...,j and T(omega) approximately 0.1 Myr. This result is discussed in connection with both the typical time scale of Earth's orbit eccentricity variation and the recent results on the typical time scale of climatic long-term variation. PMID- 12633407 TI - Comment on "Symmetrical temperature-chaos effect with positive and negative temperature shifts in a spin glass". PMID- 12633409 TI - Comment on "Universal intermediate phases of dilute electronic and molecular glasses". PMID- 12633411 TI - Comment on "Magnetic relaxation of interacting Co clusters: crossover from two- to three-dimensional lattices". PMID- 12633413 TI - Classifying N-qubit entanglement via Bell's inequalities. AB - All the states of N qubits can be classified into N-1 entanglement classes from 2 entangled to N-entangled (fully entangled) states. Each class of entangled states is characterized by an entanglement index that depends on the partition of N. The larger the entanglement index of a state, the more entangled or the less separable is the state in the sense that a larger maximal violation of Bell's inequality is attainable for this class of state. PMID- 12633414 TI - Cyclization of a polymer: first-passage problem for a non-Markovian process. AB - We discuss a closure of a polymer chain released from a state with given end-to end distance. Considering the situation as a first passage problem for a non Markovian process, we express the survival probability through the three-time distribution of the end-to-end distance and evaluate it numerically by solving a Volterra equation. We also show how the Wilemski-Fixmann approximation emerges within this scheme and discuss its geometrical nature. PMID- 12633415 TI - Quasinormal modes, the area spectrum, and black hole entropy. AB - The results of loop quantum gravity concerning geometric operators and black hole entropy are beset by an ambiguity labeled by the Immirzi parameter. We use a result from classical gravity concerning the quasinormal mode spectrum of a black hole to fix this parameter in a new way. As a result we arrive at the Bekenstein Hawking expression of A/4l(2)(P) for the entropy of a black hole and in addition see an indication that the appropriate gauge group of quantum gravity is SO(3) and not its covering group SU(2). PMID- 12633416 TI - New experimental limit on the photon rest mass with a rotating torsion balance. AB - A rotating torsion balance method is used to detect the product of the photon mass squared and the ambient cosmic vector potential A(e). The signal is modulated by rotating the torsion balance to ensure the effectiveness of detection for all possible orientations of the vector potential. The influences of sidereal disturbances of environment are also removed by virtue of this modulation method. The experimental result shows micro (2)(gamma)A(e)<1.1 x 10( 11) T m/m(2), with micro (-1)(gamma) as the characteristic length associated with photon mass. If the ambient cosmic vector potential A(e) is 10(12) T m due to cluster level fields, we obtain a new upper limit on photon mass of 1.2 x 10(-51) g. PMID- 12633418 TI - Light from cascading partons in relativistic heavy-ion collisions. AB - We calculate the production of high energy photons from Compton and annihilation processes as well as fragmentation off quarks in the parton cascade model. The multiple scattering of partons is seen to lead to a substantial production of high energy photons, which rises further when parton multiplication due to final state radiation is included. The photon yield is found to be directly proportional to the number of hard collisions and thus provides valuable information on the preequilibrium reaction dynamics. PMID- 12633420 TI - Experimental evidence for the existence of 7H and for a specific structure of 8He. AB - Experimental search for the superheavy 7H isotope was performed in the reaction p(8He,pp)7H with the 8He beam at 61.3A MeV. The evidence for existence of the 7H state near the t+4n threshold was obtained. In the same experiment, the p(8He,t) reaction populating the ground and excited 2(+) state of 6He was investigated. The obtained results argue on a specific structure of the 8He ground state containing the 6He subsystem in the excited 2(+) state with a large weight. PMID- 12633421 TI - New shape isomer in the self-conjugate nucleus 72Kr. AB - A new isomeric 0(+) state was identified as the first excited state in the self conjugate (N=Z) nucleus 72Kr. By combining for the first time conversion-electron and gamma-ray spectroscopy with the production of metastable states in high energy fragmentation, the electric-monopole decay of the new isomer to the ground state was established. The new 0(+) state is understood as the band head of the known prolate rotational structure, which strongly supports the interpretation that 72Kr is one of the rare nuclei having an oblate-deformed ground state. This observation gives in fact the first evidence for a shape isomer in a N=Z nucleus. PMID- 12633423 TI - Reaction and scattering in cold electron collisions. AB - Experimental data are presented for the scattering of electrons by CCl4 between 8 and 200 meV impact energy. These results are used in conjunction with data for the reactive process, yielding Cl-, to study the low energy behavior of a system which simultaneously displays both reactive and elastic scattering channels. Phase shifts are derived and illustrate how channel competition develops as the energy falls. This behavior and the involvement of vibronic effects at impact energies above approximately 30 meV pose a challenge to theory. PMID- 12633422 TI - Controlling the orientation of polar molecules with combined electrostatic and pulsed, nonresonant laser fields. AB - We demonstrate that molecules with a moderate permanent dipole moment can be oriented with combined electrostatic and pulsed, nonresonant laser fields. We use OCS molecules as a sample. The degree of orientation can be increased by increasing the magnitude of electrostatic field and the peak intensity of the laser field or by decreasing the rotational temperature of the molecules. PMID- 12633424 TI - Ultracold atom-atom collisions in a nonresonant laser field. AB - Using a recently developed approach for treating the three-dimensional anisotropic scattering we find considerable influence of a nonresonant laser field with intensity I> or =10(5) W/cm(2) on the Cs-Cs ultracold collisions. Strong dependence on the laser wavelength lambda(L) is shown at the optical region as lambda(L) becomes shorter than the critical value lambda(0) approximately 3000 nm (of the atomic de Broglie wave lambda) defining the region lambda(0)< or =lambda of the s-wave domination in the absence of the external field. Dependence on the laser polarization is also essential. The found effect can be applicable for controlling atom-atom interactions at ultralow temperatures. PMID- 12633425 TI - Auger recombination and charge-carrier thermalization in Hg - n -cluster photoelectron studies. AB - Photoelectron spectra (PES) of Hg(-)(n) show strong dependence of spectral features on photon energy, i.e., peak tailing and band gap filling. This dependence suggests the existence of complex photoinduced processes in parallel with the direct photodetachment process. The "corrupted" PES, taken with intermediate photon energy, carry the signature of interband absorption followed by charge-carrier thermalization and Auger electron ejection in Hg(-)(n). These processes, so significant in the photophysics of bulk semiconductors and nanoparticles, have not yet been identified in clusters. PMID- 12633426 TI - Quantum locking of mirrors in interferometers. AB - We show that quantum noise in very sensitive interferometric measurements such as gravitational-wave detectors can be drastically modified by quantum feedback. We present a new scheme based on active control to lock the motion of a mirror to a reference mirror at the quantum level. This simple technique allows one to reduce quantum effects of radiation pressure and to greatly enhance the sensitivity of the detection. PMID- 12633427 TI - Photonic band gap from a stack of positive and negative index materials. AB - Layered heterostructures combining ordinary and negative refractive index materials are shown to display a new type of photonic band gap corresponding to zero (volume) averaged refractive index. Distinct from band gaps induced by Bragg scattering, the zero-n; gap is invariant upon a change of scale length and is insensitive to disorder that is symmetric in the random variable. A metallic structure that exhibits such a band gap is explicitly designed, and its properties are calculated with accurate finite difference time domain simulations. PMID- 12633428 TI - Defect mode emission of a dye doped cholesteric polymer network. AB - We have observed enhanced fluorescence and laser emission due to a photonic defect mode in a dye doped cholesteric polymer network. The defect is caused by a phase jump of the cholesteric helix at the interface of two stacked layers of a cholesteric polymer film. Fluorescence spectra show an additional resonant mode inside the photonic stop band. Pulsed excitation gives rise to laser emission of the defect mode, with an exceptionally low lasing threshold. The defect mode emission has a circular polarization whose sense of rotation is opposite to that of the cholesteric helix. PMID- 12633429 TI - Dynamic self-assembly of rings of charged metallic spheres. AB - This Letter describes dynamic self-assembly in a system of stainless steel spheres ( approximately 1 mm in diameter) rolling on a flat dielectric surface under the influence of an external magnetic field that rotates parallel to the plane of the surface. As the spheres move, they charge triboelectrically. Self assembly is mediated by two types of electrostatic interactions among these charges: (i) attraction between negatively charged regions of the surface and positively charged spheres and (ii) repulsion between the like-charged spheres. The spheres organize into highly ordered rings as a result of these electrostatic interactions. PMID- 12633430 TI - Fano resonances with discrete breathers. AB - A theoretical study of linear wave scattering by time-periodic spatially localized excitations (discrete breathers) is presented. A peculiar effect of total reflection occurs due to a Fano resonance when a localized state originating from closed channels resonates with the open channel. For the discrete nonlinear Schrodinger chain, we give an analytical result for the frequency dependence of the transmission coefficient, including the possibility of resonant reflection. We extend the analysis to chains of weakly coupled anharmonic oscillators and discuss the relevance of the effect for electronic transport spectroscopy of mesoscopic systems. PMID- 12633419 TI - Disappearance of back-to-back high-pT hadron correlations in central Au+Au collisions at sqrt[s NN ] =200 GeV. AB - Azimuthal correlations for large transverse momentum charged hadrons have been measured over a wide pseudorapidity range and full azimuth in Au+Au and p+p collisions at sqrt[s(NN)]=200 GeV. The small-angle correlations observed in p+p collisions and at all centralities of Au+Au collisions are characteristic of hard scattering processes previously observed in high-energy collisions. A strong back to-back correlation exists for p+p and peripheral Au+Au. In contrast, the back-to back correlations are reduced considerably in the most central Au+Au collisions, indicating substantial interaction as the hard-scattered partons or their fragmentation products traverse the medium. PMID- 12633431 TI - Statistics of Lagrangian velocities in turbulent flows. AB - We present a generalized Fokker-Planck equation for the joint position-velocity probability distribution of a single fluid particle in a turbulent flow. Based on a simple estimate, the diffusion term is related to the two-point two-time Eulerian acceleration-acceleration correlation. Dimensional analysis yields a velocity increment probability distribution with normal scaling v approximately t(1/2). However, the statistics need not be Gaussian. PMID- 12633432 TI - Particle energization in an expanding magnetized relativistic plasma. AB - Using a 21 / 2-dimensional particle-in-cell (PIC) code to simulate the relativistic expansion of a magnetized collisionless plasma into a vacuum, we report a new mechanism in which the magnetic energy is efficiently converted into the directed kinetic energy of a small fraction of surface particles. We study this mechanism for both electron-positron and electron-ion (m(i)/m(e)=100, m(e) is the electron rest mass) plasmas. For the electron-positron case, the pairs can be accelerated to ultrarelativistic energies. For electron-ion plasmas, most of the energy gain goes to the ions. PMID- 12633433 TI - Slow magnetosonic solitons detected by the cluster spacecraft. AB - Experimental evidence is provided for the existence of slow-mode magnetosonic solitons in the col-lisionless plasma at the magnetopause boundary layer. The solitons were detected by the fleet of Cluster spacecraft at the dusk flank of the magnetosphere as magnetic field depressions (up to 85%) accom-panied with enhancement of the plasma density and temperature by a factor of 2. The solitons propagate 250 km/s with respect to the satellites and have perpendicular size of 1000-2000 km, which is a few ion inertial scale lengths. The comparison with numerical solutions of a theoretical model shows quantitative agreement between the model and observations. PMID- 12633434 TI - Fluctuations of the phase difference across an array of Josephson junctions in superfluid 4He near the lambda Transition. AB - We present a formal thermodynamic treatment of superfluid flow in a Josephson junction. We show that the current i(s) and the phase difference phi are thermodynamic conjugate variables. We derive quantitative expressions for the rms fluctuations of i(s) and phi. Also, we discuss the thermodynamic stability and the thermal activation to the phase-slip region. We apply the developed formalism to show why an array of apertures in 4He can exhibit the Josephson effect near the lambda transition despite strong thermal fluctuations. PMID- 12633435 TI - Excitation energies and radiative lifetimes of Ge1-xSix nanocrystals: alloying versus confinement effects. AB - The composition dependence of the optical and structural properties of Ge1-xSix nanocrystals is investigated by means of ab initio total-energy and electronic structure calculations. A trimodal distribution of the Ge-Ge, Ge-Si, and Si-Si bond lengths is found. The pair-excitation energies and the Stokes shift are calculated taking into account many-body and alloying effects. They show a distinct nonlinear behavior with changing composition. The radiative lifetime decreases exponentially with increasing Ge molar fraction. The theoretical results explain recent photoluminescence measurements. They show that composition and confinement effects can be discussed nearly separately. PMID- 12633436 TI - Chemical- or radiation-assisted selective dealloying in bimetallic nanoclusters. AB - A selective dealloying in bimetallic nanoclusters prepared by ion implantation has been found upon thermal annealing in oxidizing atmosphere or irradiation with light ions. In the first process, the incoming oxygen interacts preferentially with copper promoting Cu2O formation, therefore extracting copper from the alloy. In the second process the irradiation with Ne ions promotes a preferential extraction of Au from the alloy, resulting in the formation of Au-enriched "satellite" nanoparticles around the original AuxCu1-x cluster. PMID- 12633437 TI - Anchoring-mediated interaction of edge dislocations with bounding surfaces in confined cholesteric liquid crystals. AB - We employ the fluorescent confocal polarizing microscopy to image edge dislocations in cholesteric liquid crystals. Surface anchoring at the bounding plates determines the structure and behavior of defects. Two types of plates set in-plane director orientation but differ in the type of associated anchoring potentials. Plates with strong polar anchoring and nonzero azimuthal anchoring repel the dislocations, while plates with weak polar anchoring and no azimuthal anchoring allow the dislocations to escape through the boundary. To explain the results, we propose a coarse-grained model of cholesteric anchoring. PMID- 12633417 TI - Search for a W' boson decaying to a top and bottom quark pair in 1.8 TeV pp collisions. AB - We report the results of a search for a W' boson produced in pp; collisions at a center-of-mass energy of 1.8 TeV using a 106 pb(-1) data sample recorded by the Collider Detector at Fermilab. We observe no significant excess of events above background for a W' boson decaying to a top and bottom quark pair. In a model where this boson would mediate interactions involving a massive right-handed neutrino (nu(R)) and have standard model strength couplings, we use these data to exclude a W' boson with mass between 225 and 536 GeV/c(2) at 95% confidence level for M(W')>>M(nu(R)) and between 225 and 566 GeV/c(2) at 95% confidence level for M(W')160% of pTEE (+/-2 SD) and <70% or >130% of pTEE (+/-1 SD), respectively. Use of only the +/-1 SD cut-offs, not the +/-2 SD cut-offs, resulted in a relationship between rEI and body weight similar to what was expected (based on an independently calculated relationship between rEI and measured TEE). Exclusion of rEI outside either the +/-2 SD (11% of subjects) or +/-1 SD (57% of subjects) cut-offs did not affect mean reported macronutrient intakes, but did markedly affect relationships between dietary composition and body mass index. CONCLUSIONS: When examining relationships between diet and health, use of +/-1 SD cut-offs may be preferable to +/-2 SD cut offs for excluding inaccurate dietary reports. PMID- 12633512 TI - Assessing the dietary environment: examples from school-based nutrition interventions. AB - OBJECTIVE: The objectives of this paper are to (1) introduce the concept of upstream and downstream public health approaches and discuss diet assessment issues in that context, and (2) provide examples of diet assessment methods and challenges in assessing environmental factors influencing eating patterns. DESIGN: Dietary assessment of environmental factors is discussed as they relate to nutrition interventions for school-aged children, although the issues transcend population characteristics. Examples of assessment challenges in measuring 'dietary environments' are drawn from the Child and Adolescent Trial for Cardiovascular Health (CATCH), Teens Eating for Energy and Nutrition at School (TEENS) and Trying Alternative Cafeteria Options in Schools (TACOS) studies. RESULTS/CONCLUSIONS: The healthfulness of our 'dietary environments' (which may include food availability, social norms around food choice and the effect of pricing, policy and promotion on food choice) may be more important in determining what people consume than their individual decision-making about food choice. There is a dearth of published information to inform us on how to assess these 'dietary environments'. PMID- 12633513 TI - The SENECA study: potentials and problems in relating diet to survival over 10 years. AB - OBJECTIVE: To give an overview of the evaluation of the modified diet history applied in the SENECA study (Survey in Europe on Nutrition and the Elderly; a Concerted Action). DESIGN: Nineteen centres in 12 countries participated at baseline. Nine of these SENECA centres conducted a repeat measurement scheme in elderly people born between 1913 and 1918. These longitudinal centres included 100 subjects per sex per site. METHODS: The relative validity of the method was tested by comparing results of the modified diet history with results obtained from a weighed record in 82 subjects. In the follow-up we compared changes in energy intake with changes in body weight and calculated the physical activity ratio in all longitudinal centres. In SENECA's finale we examined the predictive value of dietary patterns observed at baseline for survival 10 years later, making use of the original and an adapted Mediterranean Diet Score. RESULTS: The modified diet history overestimated intake, compared with the weighed record. However, the physical activity ratio and an in-depth study in a metabolic room indicated that the diet history rather underestimated energy intake. We did not find a relationship between changes in energy intake and changes in body weight, but this could be explained by the fact that most likely we did not measure intake in the dynamic phase of body weight change. Based on the adapted Mediterranean Diet Score, the study results showed a positive relation between a healthy diet and survival. CONCLUSION: We conclude that the modified diet history has given sufficiently reliable results for the purposes of the study. PMID- 12633514 TI - Measuring the intakes of foods and nutrients of marginal populations in north west Mexico. AB - OBJECTIVE: To describe methodological issues regarding the measurement of food intake of marginal populations in the north-west of Mexico. METHODS: Dietary data from three sources were analysed. The Sonora State Food Basket Study (505 individuals from all ages, both sexes, high, medium and low income, and from urban and rural settings), 144 individuals from rural Indian communities, and 152 low-income urban women. Methods of dietary data collection were single 24-hour recalls for the first two studies, and four non-consecutive 24-hour recalls as well as a food-frequency questionnaire (FFQ) for low-income urban women. RESULTS: The food patterns in the three studies showed similar core foods; however, food diversity resulted in 19, 13 and 57 different foods reported by 90% of the population in the state, Indian communities and urban women, respectively. Mean intakes of selected food components in the sample of urban women vs. a representative sample of the population of the state were similar, although smaller variation in consumption was observed in urban women with repeated 24 hour recalls. Inter-individual variation in intake of food components was similar in the state and urban women samples, but the Indian communities showed lower coefficients of variation (CVs). Use of four 24-hour recalls decreased the inter individual variation in food component intakes of low-income urban women, especially for vitamin intakes. The FFQ did not show an additional decrease in inter-individual variation for macronutrients, but reduced even more the CVs for vitamins. Intra-individual variation was higher than inter-individual variation for urban women's intakes when estimated by repeated 24-hour recalls. This effect was reversed when estimated by duplicate food frequencies. Ratios of intra individual to inter-individual variation in food intake were lower for Mexican when compared with US women. These results should be considered, especially for association analysis of low-income women's diets and health outcomes. PMID- 12633515 TI - 'Malnutrition' in the aged: the dietary assessment. AB - Changing demography towards older populations requires nutritionists to review food--health relationships in regard to older age groups, to the ageing process, to precursors of disability and survival, and to the changeability of health status through diet in later life. Nutritional status in later life may be complex with combined features of under-, over- and disordered nutrition -- dysnutrition. Further, beyond food components (essential nutrients and more), foods themselves, integrals of food intake and the socio-cultural aspects of food intake play a significant role in the health and well-being of the aged and require measurement. Ageing socio-biology provides opportunities and imposes limitations on the usual food intake instruments. However, rapid assessment procedures can be particularly valuable as highlighted in the Food Habits in Later Life project of the International Union of Nutritional Sciences. There are critical issues to which dietary methodology can be applied to the aged: a disappearing knowledge base, relevant health-enhancing strategies and the creation of new food--health opportunities. Many communities now wish to use such information to develop food-based dietary guidelines for their aged members. PMID- 12633516 TI - Bias in dietary-report instruments and its implications for nutritional epidemiology. AB - OBJECTIVE: To evaluate measurement error structure in dietary assessment instruments and to investigate its implications for nutritional studies, using urinary nitrogen excretion as a reference biomarker for protein intake. DESIGN: The dietary assessment methods included different food-frequency questionnaires (FFQs) and such conventional dietary-report reference instruments as a series of 24-hour recalls, 4-day weighed food records or 7-day diaries. SETTING: Six original pilot validation studies within the European Prospective Investigation of Cancer (EPIC), and two validation studies conducted by the British Medical Research Council (MRC) within the Norfolk cohort that later joined as a collaborative component cohort of EPIC. SUBJECTS: A sample of approximately 100 to 200 women and men, aged 35-74 years, from each of eight validation studies. RESULTS: In assessing protein intake, all conventional dietary-report reference methods violated the critical requirements for a valid reference instrument for evaluating, and adjusting for, dietary measurement error in an FFQ. They displayed systematic bias that depended partly on true intake and partly was person-specific, correlated with person-specific bias in the FFQ. Using the dietary-report methods as reference instruments produced substantial overestimation (up to 230%) of the FFQ correlation with true usual intake and serious underestimation (up to 240%) of the degree of attenuation of FFQ-based log relative risks. CONCLUSION: The impact of measurement error in dietary assessment instruments on the design, analysis and interpretation of nutritional studies may be much greater than has been previously estimated, at least regarding protein intake. PMID- 12633517 TI - Measuring flavonoid intake: need for advanced tools. AB - OBJECTIVE: Flavonoids are phytochemicals with potentially beneficial biological effects that are poorly characterised in existing tables of food composition. DESIGN: To describe new techniques for analysis, absorption, informatics and dietary assessment that are important in measuring the flavonoid content in foods and in developing a flavonoid food composition database. SETTING: Data on chemical analyses of the major flavonoid compounds that exist in the food science literature are being located, collated and compiled into a preliminary flavonoid food composition database. RESULTS: The analytical process begins with preparation of the food for flavonoid analysis using techniques to disrupt the food matrix, alcoholic extraction, and enzymatic or acidic hydrolysis to remove sugars. Separation is usually accomplished using high-performance liquid chromatography. Flavonoids are identified by diode array spectrometry, mass spectrometry or nuclear magnetic resonance spectroscopy. Quantification usually employs comparison of the sample with standards, if available, using the area under the curve of the chromatogram to estimate quantity. Absorption studies are useful since flavonoids vary in their absorption. Finally, information management technologies (informatics) are used to translate flavonoid data information into food composition databases. This process involves identification of foods containing the compounds of interest, collection and organisation of sources of existing analytical data, assignment of quality scores or aggregation of acceptable data for each component and food, calculation of appropriate statistics, assignment of food codes and verification, and finalization. The resulting food flavonoid database can be used with state-of-the-art dietary assessment methods to develop estimates of flavonoid intakes in foods and to correlate these with estimates of disease risk. CONCLUSIONS: A provisional flavonoid database, including at least two components from each of six classes of flavonoids, should be completed in 2002. PMID- 12633518 TI - Erythrocyte cell membrane phospholipid levels compared against reported dietary intakes of polyunsaturated fatty acids in pregnant Mexican women. AB - OBJECTIVE: To evaluate the validity of a food-frequency questionnaire (FFQ) for assessment of the dietary intakes of polyunsaturated fatty acids (PUFAs) against a biochemical marker of fat intake, erythrocyte cell membrane phospholipid levels, during pregnancy. DESIGN: Cross-sectional analysis. SETTING: Developmental Neurobiology Department, National Institute of Perinatology, Mexico City. SUBJECTS: One hundred forty-six healthy pregnant women during the last trimester of pregnancy. Among women enrolled, the first 35 pregnant women (24%) had their erythrocytes analysed for fatty acid status. METHODS: We administered an FFQ and compared intakes of PUFAs against their erythrocyte cell membrane concentrations, processed by gas chromatography. RESULTS: Pearson correlation coefficients among alpha-linolenic acid (ALN), docosahexaenoic acid (DHA) and eicosapentaenoic acid in erythrocyte cell membranes against their crude dietary counterparts were 0.32, 0.35 and 0.36 (each P < 0.05). In a simple linear regression, erythrocyte DHA and arachidonic acid (AA) were significantly related to their respective dietary intakes (beta = 0.30, 95% confidence interval (CI): 0.007-0.60, P=0.045 for DHA; beta = 0.49, 95% CI: 0.010-0.98, P=0.044 for AA). Erythrocyte cell membrane ALN concentration (%/total) was only marginally related to ALN dietary intake (mg day-1) (beta = 0.52, 95% CI: -0.020-1.10, P=0.061). However, after adjustment for long-chain n-3 PUFA/AA, this association reached significance (beta = 0.44, 95% CI: 0.026-0.825, P=0.038). Main dietary sources for n-3 PUFAs were canned tuna fish and fresh catfish; for n-6 these were eggs and cow's milk. The use of this FFQ in these pregnant Mexican women provided estimates of average long-term intakes of PUFAs and correlated reasonably well with their erythrocyte cell membrane phospholipid status. However, we need to consider that, during pregnancy, there is a faster turnover of PUFAs from fat storage that may modify the profile of erythrocyte PUFAs and lower the correlation between dietary intake and erythrocyte PUFAs. PMID- 12633519 TI - Development of a questionnaire and a database for assessing dietary d-limonene intake. AB - OBJECTIVE: Increasing recognition of the potential importance of phytochemicals in the aetiology of cancer and heart diseases has highlighted the need for methods to measure individual phytochemical consumption that are sufficiently simple to be used in large epidemiological studies and whose reproducibility and accuracy have been quantified. d-Limonene is a natural component of a variety of foods and beverages and is found mainly in citrus fruits. However, d-limonene is not assessed by any nationally available analysis database. DESIGN: We designed our study to assess the d-limonene content of different citrus juices and beverages and to develop a dietary assessment instrument to measure consumption of citrus foods (fruit, juice and peel) and d-limonene intake and test it for reliability. SUBJECTS AND METHODS: A total of 120 citrus juice samples were analysed and used to develop the preliminary d-limonene database. A self administered citrus food-frequency questionnaire was developed and administered twice to participants, separated by a 2-month interval. The questionnaire was tested for reproducibility of estimates of citrus food consumption and d-limonene intake among 120 participants. RESULTS: Correlation coefficients between the two administrations of the questionnaire ranged from 0.50 for citrus peel use to 0.82 for orange juice. Mean intakes (range) of d-limonene from citrus juices among consumers were 13.0 (0.24-141.9) mg day-1 and 13.2 (0.07-83.9) mg day-1 (r = 0.60, P < 0.001). CONCLUSION: The citrus frequency questionnaire developed in this study provided highly reproducible estimates of citrus foods, citrus peel and d-limonene intakes. This instrument may be a useful tool in studies of the associations between citrus peel use, d-limonene intake and risk for chronic disease. PMID- 12633520 TI - Understanding the nutrition transition: measuring rapid dietary changes in transitional countries. AB - OBJECTIVE: To understand methodological concerns related to dietary intake collection in transitional societies. DESIGN: Three days of household weighing and measurement of all food used and repeated 24-hour recalls. SETTING: Eight Chinese provinces. SUBJECTS: Five thousand nine hundred and fifty-two and 5152 adults aged 20-45 years in 1989 and 1997, respectively. RESULTS: Great variance exists in the types and quantity of animal products used in common recipes. For example, the proportion of pork from lean cuts in 'stir-fried fresh pepper and pork' varies between 14 and 24% in urban and rural areas and the total pork content for 100 g (dish) varies by 15 to 19 g between rural and urban areas in each of eight provinces. Another challenge relates to the variation in the edible vegetable oil content added during food preparation. Reliance on standard recipes for each fried dish would miss the variations in oil use over time, space and socio-economic status. CONCLUSIONS: Dietary change is rapid in transitional countries. Reliance on recipes standardised for animal food and edible oil contents will lead to very large systematic errors in the measurement of energy, fat and protein intakes. PMID- 12633521 TI - Dietary assessment tools for developing countries for use in multi-centric, collaborative protocols. AB - In recent years, increasing interest in the format of multi-centric studies among different populations in developing nations has evolved in the field of health surveys and epidemiology. Dietary intake data are most often part of these cross cultural and cross-national collaborative efforts. Various questions have been raised about the appropriate endeavours for dietetics and nutritional sciences in developing societies, the instruments available for application, and the pitfalls and caveats in their use. An important consideration is that studies be hypothesis-driven and not mere 'fishing expeditions' of unfocused data gathering. All known dietary intake measurement tools are within the preview of developing country research, but they often must be adapted individually and differentially to suit a given population. In a multi-centric context, this is complicated. The watchword should be collecting comparable information across sites, not using identical approaches. Choice of dietary intake measurement tools must be honed to the hypotheses and assumptions, on the one hand, and the exigencies and pitfalls of working in the developing country milieu, in which linguistics, seasonality, migration, uncommonness of food systems and ethical considerations present barriers and caveats, on the other. Within the hypotheses, the assumptions regarding the penetration of the measured exposures must be borne in mind. Multi centre studies in developing countries have relevance and importance in the context of food security, diet and disease, eating behaviour and satiety regulation, and nutritional anthropology. PMID- 12633522 TI - Research strategies and the use of nutrient biomarkers in studies of diet and chronic disease. AB - OBJECTIVE: To provide an account of the state of diet and chronic disease research designs and methods; to discuss the role and potential of aggregate and analytical observational studies and randomised controlled intervention trials; and to propose strategies for strengthening each type of study, with particular emphasis on the use of nutrient biomarkers in cohort study settings. DESIGN: Observations from diet and disease studies conducted over the past 25 years are used to identify the strengths and weaknesses of various study designs that have been used to associate nutrient consumption with chronic disease risk. It is argued that a varied research programme, employing multiple study designs, is needed in response to the widely different biases and constraints that attend aggregate and analytical epidemiological studies and controlled intervention trials. Study design modifications are considered that may be able to enhance the reliability of aggregate and analytical nutritional epidemiological studies. Specifically, the potential of nutrient biomarker measurements that provide an objective assessment of nutrient consumption to enhance analytical study reliability is emphasised. A statistical model for combining nutrient biomarker data with self-report nutrient consumption estimates is described, and related ongoing work on odds ratio parameter estimation is outlined briefly. Finally, a recently completed nutritional biomarker study among 102 postmenopausal women in Seattle is mentioned. The statistical model will be applied to biomarker data on energy expenditure, urinary nitrogen, selected blood fatty acid measurements and various blood micronutrient concentrations, and food frequency self-report data, to identify study subject characteristics, such as body mass, age or socio economic status, that may be associated with the measurement properties of food frequency nutrient consumption estimates. This information will be crucial for the design of a potential larger nutrient biomarker study within the cohort study component of the Women's Health Initiative. SETTING AND SUBJECTS: The methodology under study is expected to be pertinent to a wide variety of diet and chronic disease association studies in the general population. Ongoing work focuses on statistical methods developed using computer simulations motivated by studies of dietary fat in relation to breast and colon cancer among post-menopausal women, and ongoing pilot studies to be described in detail elsewhere, involving post menopausal women living in the Seattle area. RESULTS AND CONCLUSION: A varied research programme appears to be needed to make progress in the challenging diet and chronic disease research area. Such progress may include aggregate studies of diet and chronic disease that include sample surveys in diverse population groups world-wide, analytical epidemiological studies that use nutrient biomarker data to calibrate self-report nutrient consumption estimates, and randomised controlled intervention trials that arise from an enhanced infrastructure for intervention development. New innovative designs, models and methodologies are needed for each such research setting. PMID- 12633523 TI - Using national dietary data to measure dietary changes. AB - OBJECTIVE: To demonstrate that dietary datasets from the Continuing Survey of Food Intakes by Individuals, a US population survey, allow comparisons with national data and provide food composition datasets that can be used to generate similar dietary data. DESIGN: Two studies are described: the Lower Mississippi Delta Nutrition Intervention Research Initiative (Delta NIRI), which used a 24 hour recall, and a Department of Defense Military Nutrition Research Task, which used 3-day dietary records. Both studies used the same food composition tables. SETTING: Rural Lower Mississippi Delta and an Army post. SUBJECTS: Four hundred and nine residents (adults and children) from the rural Delta region of Arkansas, Louisiana and Mississippi, and 74 career soldiers from the Sergeants Major Academy, Fort Bliss, Texas. RESULTS: The Delta NIRI study found that fruit and vegetable consumption for these rural residents was lower than that found nationally. Additionally, the quality of vegetable servings is of concern since a large percentage came from french fries and potato chips. In the Sergeants Major Academy study, the national survey food composition tables allowed for easy analysis of intake data and comparisons with dietary recommendations. CONCLUSIONS: Strategies similar to those used for the Delta NIRI and Military Nutrition Research Task can be used widely, allowing comparisons of 'defined populations' with nationally distributed data. Additionally, measurement of dietary change is more efficient when the same protocol is used subsequently to collect more data, a method similar to that used by the US Department of Agriculture to describe food consumption patterns from one survey to another. PMID- 12633524 TI - Issues in the development and use of food composition databases. AB - OBJECTIVES: To share experience on the development of national and regional food composition tables (FCTs) and the issues and problems involved. DESIGN: Food composition data analysed at national and regional levels were compiled. Systematic development of FCTs detailing specific criteria and characteristics following the INFOODS and ASEANFOODS guidelines was conducted. SETTING: The national FCT was developed using the Thai FCT as a study model. For the regional level, the Association of South East Asian Nations (ASEAN) FCT was developed. SUBJECTS: About 3000 analysed datasets of about 1000 food items on proximate composition, minerals, vitamins and other available components from various sources in Thailand and about 4500 analysed datasets of 1700 food items on the same components from various countries in ASEAN were compiled. The national and regional FCTs were then developed systematically. RESULTS: The developed Thai FCT contains 24 nutrients of 1055 food items and the ASEAN FCT contains 21 nutrients of about 1740 food items. Issues and problems regarding the development and use of national and regional food composition data are listed. CONCLUSION: Food composition databases have become increasingly important in the international health arena. Systematic compilation of the data is necessary and the use of INFOODS tag names is strongly recommended. This is to indicate the methods of nutrient analysis and calculation, and to facilitate international and regional data interchange. Users are also recommended to read the Explanatory Notes and Information to the Users. Collaboration among data generators, users and compilers at national and regional levels, as well as support from international organisations, is essential. PMID- 12633526 TI - A systematic review and evaluation of the use of tumour markers in paediatric oncology: Ewing's sarcoma and neuroblastoma. PMID- 12633525 TI - A multicentre randomised controlled trial assessing the costs and benefits of using structured information and analysis of women's preferences in the management of menorrhagia. PMID- 12633527 TI - A review of the clinical effectiveness and cost-effectiveness of routine anti-D prophylaxis for pregnant women who are rhesus-negative. PMID- 12633528 TI - A systematic review of the costs and effectiveness of different models of paediatric home care. PMID- 12633529 TI - The effectiveness and cost-effectiveness of imatinib in chronic myeloid leukaemia: a systematic review. PMID- 12633530 TI - Bayesian estimation in animal breeding using the Dirichlet process prior for correlated random effects. AB - In the case of the mixed linear model the random effects are usually assumed to be normally distributed in both the Bayesian and classical frameworks. In this paper, the Dirichlet process prior was used to provide nonparametric Bayesian estimates for correlated random effects. This goal was achieved by providing a Gibbs sampler algorithm that allows these correlated random effects to have a nonparametric prior distribution. A sampling based method is illustrated. This method which is employed by transforming the genetic covariance matrix to an identity matrix so that the random effects are uncorrelated, is an extension of the theory and the results of previous researchers. Also by using Gibbs sampling and data augmentation a simulation procedure was derived for estimating the precision parameter M associated with the Dirichlet process prior. All needed conditional posterior distributions are given. To illustrate the application, data from the Elsenburg Dormer sheep stud were analysed. A total of 3325 weaning weight records from the progeny of 101 sires were used. PMID- 12633531 TI - Multivariate Bayesian analysis of Gaussian, right censored Gaussian, ordered categorical and binary traits using Gibbs sampling. AB - A fully Bayesian analysis using Gibbs sampling and data augmentation in a multivariate model of Gaussian, right censored, and grouped Gaussian traits is described. The grouped Gaussian traits are either ordered categorical traits (with more than two categories) or binary traits, where the grouping is determined via thresholds on the underlying Gaussian scale, the liability scale. Allowances are made for unequal models, unknown covariance matrices and missing data. Having outlined the theory, strategies for implementation are reviewed. These include joint sampling of location parameters; efficient sampling from the fully conditional posterior distribution of augmented data, a multivariate truncated normal distribution; and sampling from the conditional inverse Wishart distribution, the fully conditional posterior distribution of the residual covariance matrix. Finally, a simulated dataset was analysed to illustrate the methodology. This paper concentrates on a model where residuals associated with liabilities of the binary traits are assumed to be independent. A Bayesian analysis using Gibbs sampling is outlined for the model where this assumption is relaxed. PMID- 12633532 TI - Use of the score test as a goodness-of-fit measure of the covariance structure in genetic analysis of longitudinal data. AB - Model selection is an essential issue in longitudinal data analysis since many different models have been proposed to fit the covariance structure. The likelihood criterion is commonly used and allows to compare the fit of alternative models. Its value does not reflect, however, the potential improvement that can still be reached in fitting the data unless a reference model with the actual covariance structure is available. The score test approach does not require the knowledge of a reference model, and the score statistic has a meaningful interpretation in itself as a goodness-of-fit measure. The aim of this paper was to show how the score statistic may be separated into the genetic and environmental parts, which is difficult with the likelihood criterion, and how it can be used to check parametric assumptions made on variance and correlation parameters. Selection of models for genetic analysis was applied to a dairy cattle example for milk production. PMID- 12633533 TI - Genetic parameters for resistance to the Salmonella abortusovis vaccinal strain Rv6 in sheep. AB - An experimental population (1216 lambs from 30 sires) of the Inra401 sheep was created in an Inra flock to allow QTL detection for susceptibility to Salmonella infection, wool and carcass traits. The Inra401 is a sheep composite line developed from two breeds: Berrichon du Cher and Romanov. At 113 days of age on average, the lambs were inoculated intravenously with 10(8) Salmonella abortusovis Rv6 (vaccinal strain). They were slaughtered 10 days after the inoculation. Several traits were measured at inoculation and/or slaughtering to estimate the genetic resistance of the lambs to Salmonella infection: specific IgM and IgG1 antibody titres, body weight loss, spleen and pre-scapular node weights and counts of viable Salmonella persisting in these organs. This paper presents a quantitative analysis of the genetic variability of the traits related to salmonellosis susceptibility. The heritabilities of the traits varied between 0.10 and 0.64 (significantly different from zero). Thus, in sheep as well as in other species, the determinism of resistance to Salmonella infection is under genetic control. Moreover, the correlations between the traits are in agreement with the known immune mechanisms. The genetic variability observed should help QTL detection. PMID- 12633534 TI - Estimation of genetic variability and selection response for clutch length in dwarf brown-egg layers carrying or not the naked neck gene. AB - In order to investigate the possibility of using the dwarf gene for egg production, two dwarf brown-egg laying lines were selected for 16 generations on average clutch length; one line (L1) was normally feathered and the other (L2) was homozygous for the naked neck gene NA. A control line from the same base population, dwarf and segregating for the NA gene, was maintained during the selection experiment under random mating. The average clutch length was normalized using a Box-Cox transformation. Genetic variability and selection response were estimated either with the mixed model methodology, or with the classical methods for calculating genetic gain, as the deviation from the control line, and the realized heritability, as the ratio of the selection response on cumulative selection differentials. Heritability of average clutch length was estimated to be 0.42 +/- 0.02, with a multiple trait animal model, whereas the estimates of the realized heritability were lower, being 0.28 and 0.22 in lines L1 and L2, respectively. REML estimates of heritability were found to decline with generations of selection, suggesting a departure from the infinitesimal model, either because a limited number of genes was involved, or their frequencies were changed. The yearly genetic gains in average clutch length, after normalization, were estimated to be 0.37 +/- 0.02 and 0.33 +/- 0.04 with the classical methods, 0.46 +/- 0.02 and 0.43 +/- 0.01 with animal model methodology, for lines L1 and L2 respectively, which represented about 30% of the genetic standard deviation on the transformed scale. Selection response appeared to be faster in line L2, homozygous for the NA gene, but the final cumulated selection response for clutch length was not different between the L1 and L2 lines at generation 16. PMID- 12633535 TI - Substitution of the alpha-lactalbumin transcription unit by a CAT cDNA within a BAC clone silenced the locus in transgenic mice without affecting the physically linked Cyclin T1 gene. AB - We recently reported that a goat bacterial artificial chromosome (BAC) clone conferred site-independent expression in transgenic mice of the two loci present within its insert, the ubiquitously expressed Cyclin T1 and the mammary specific alpha-lactalbumin (alphalac) genes. To assess if this vector could target mammary restricted expression of cDNA, the CAT ORF was introduced by homologous recombination in Escherichia coli in place of the alphalac transcription unit. The insert of this modified BAC was injected into mice and three transgenic lines were derived. None of these lines expressed the CAT gene suggesting that the use of long genomic inserts is not sufficient to support the expression of intron less transgenes. The physically linked goat Cyclin T1 locus was found to be active in all three lines. This observation reinforced the hypothesis that the two loci are localised in two separate chromatin domains. PMID- 12633536 TI - A mutation in the LAMC2 gene causes the Herlitz junctional epidermolysis bullosa (H-JEB) in two French draft horse breeds. AB - Epidermolysis bullosa (EB) is a heterogeneous group of inherited diseases characterised by skin blistering and fragility. In humans, one of the most severe forms of EB known as Herlitz-junctional EB (H-JEB), is caused by mutations in the laminin 5 genes. EB has been described in several species, like cattle, sheep, dogs, cats and horses where the mutation, a cytosine insertion in exon 10 of the LAMC2 gene, was very recently identified in Belgian horses as the mutation responsible for JEB. In this study, the same mutation was found to be totally associated with the JEB phenotype in two French draft horse breeds, Trait Breton and Trait Comtois. This result provides breeders a molecular test to better manage their breeding strategies by genetic counselling. PMID- 12633537 TI - Oral contraceptives and the risk of myocardial infarction. PMID- 12633538 TI - Looking for people at high cardiovascular risk? Look at serum-creatinine. PMID- 12633539 TI - Multi-chamber pacing: a perfect solution for cardiac mechanical dyssynchrony? PMID- 12633540 TI - Blood pressure lowering and ACE inhibition for the avoidance of cardiac and cerebral events. PMID- 12633541 TI - Direct stenting. AB - Stents have ameliorated the outcome of percutaneous coronary interventions. Improved design, profile and flexibility of the currently available stents now permit to deliver the stent without pre-dilatation. In western European countries, stent implantation now occurs in up to 80% of all percutaneous revascularization procedures. In selected cases the safety and feasibility of direct stenting over conventional balloon-predilatation stenting has been demonstrated, in association with slightly superior outcome. This likely results from reduced vessel wall damage-dissections are sealed at the moment they are created-and distal embolization is reduced. In addition, a significant reduction in terms of costs and duration of procedures was demonstrated, while restenosis rate is not reduced with direct stenting. PMID- 12633542 TI - Progenitor and embryonic stem cell transplantation for myocardial angiogenesis and functional restoration. PMID- 12633543 TI - Accelerated decline and prognostic impact of renal function after myocardial infarction and the benefits of ACE inhibition: the CATS randomized trial. AB - AIMS: Information regarding the cardiorenal axis in patients after a myocardial infarction (MI) is limited. We examined the change in renal function after a first MI, the protective effect of angiotensin converting enzyme (ACE) inhibition and the prognostic value of baseline renal function. METHODS AND RESULTS: The study population consisted of 298 patients with a first anterior wall MI who were randomized to the ACE inhibitor captopril or placebo after completion of streptokinase infusion. Renal function, by means of glomerular filtration rate (GFR), was calculated using the Cockroft-Gault equation (GFR(c)). In the placebo group, renal function (GFR(c)) declined by 5.5 min(-1)within 1 year, vs only 0.5 ml min(-1)in the ACE inhibitor group (P<0.05). This beneficial effect of captopril was most pronounced in patients with the most compromised renal function at baseline. The incidence of chronic heart failure (CHF) within 1 year increased significantly with decreasing GFR(c)(divided into tertiles: 24.0, 28.9, and 41.2%; P<0.01). The risk-ratio for GFR(c)<81 ml min(-1)vs >103 mL min(-1)was 1.86 (95% CI 1.11-3.13; P=0.019). CONCLUSIONS: Renal function markedly deteriorates after a first MI, but is significantly preserved by ACE inhibition. Furthermore, an impaired baseline renal function adds to the prognostic risk of developing CHF in patients after a first anterior MI. PMID- 12633544 TI - Direct coronary stent implantation does not reduce the incidence of in-stent restenosis or major adverse cardiac events: six month results of a randomized trial. AB - STUDY OBJECTIVE: To compare the long-term angiographic, clinical and economic outcome of direct stenting vs stenting after balloon predilatation. PATIENT POPULATION AND METHODS: Four hundred patients with coronary stenoses in a single native vessel were randomized to direct stenting vs stenting after predilatation. A major adverse cardiac and cerebral event (MACCE) was defined as death, myocardial infarction, stent thrombosis, target restenosis, repeat target- and non-target vessel-related percutaneous coronary intervention, target lesion revascularization, coronary artery bypass surgery and stroke. RESULTS: Stents were successfully implanted in 98.3% of patients randomized to direct stenting vs 97.8% randomized to stenting preceded by predilatation. The primary success rate of direct stenting was 88.3%, vs 97.8% for stenting preceded by balloon dilatation (P=0.01). The angiographic follow-up at 6 months included 333 of the 400 patients (83%). The binary in-stent restenosis rate was 23.1% of 163 patients randomized to direct stenting vs 18.8% of 166 patients randomized to balloon predilatation (P=0.32). By 185+/-25 days, MACCE had occurred in 31 of 200 (15.5%) patients randomized to direct stenting, vs 33 of 200 (16.5%) randomized to predilatation (P=0.89). At 6 months, costs associated with the direct stenting strategy (Euros 3222/patient) were similar to those associated with predilatation (Euros 3428/patient, P=0.43). However, procedural costs were significantly lower. It is noteworthy that, on multivariate analysis, a baseline C-reactive protein level >10 mg l(-1)was a predictor of restenosis (odds ratio: 2.10, P=0.025) as well as of MACCE (odds ratio: 1.94, P=0.045). CONCLUSIONS: Compared to stenting preceded by balloon predilatation, direct stenting was associated with similar 6 month restenosis and MACCE rates. Procedural, but not overall 6-month costs, were reduced by direct stenting. An increased baseline CRP level was an independent predictor of adverse long-term outcome after coronary stent implantation. PMID- 12633545 TI - Left ventricular remodelling and haemodynamic effects of multisite biventricular pacing in patients with left ventricular systolic dysfunction and activation disturbances in sinus rhythm: sub-study of the MUSTIC (Multisite Stimulationin Cardiomyopathies) trial. AB - AIMS: To use echocardiography to determine early and late haemodynamic effects of atrio-biventricular (A-BiV) pacing on left ventricular (LV) function and their interrelations with exercise tolerance. METHODS: Thirty-four patients with ejection fraction <35% (18 idiopathic dilated cardiomyopathy (DCM) and 16 ischaemic cardiomyopathy, in sinus rhythm and with intra-ventricular conduction delay (IVCD)) were implanted with transvenous A-BiV pacemakers. Echocardiographic measurements were compared before implantation, after 3 months A-BiV pacing, and 3 months inactive pacing (ventricular inhibited pacing at 40 beats x min(-1)as part of a crossover design, and at 9- and 12-month longitudinal follow-up. RESULTS: Total isovolumic time (IVT) halved after 3 months A-BiV pacing (from 20.1+/-4.4 to 10.7+/-4.9s min(-1), P<0.001) and did not change thereafter. LV cavity size fell after 3 months (end-diastolic dimension (EDD) 7.3+/-0.8 to 6.8+/ 0.8 cm, P<0.001, and end-systolic dimension (ESD) 6.2+/-0.8 to 5.9+/-0.8 cm, P<0.05), with a further fall in EDD and ESD (by 8.4+/-7.8 and 8.8+/-7.8mm, respectively, both P<0.001) after 12 months. Although not a primary end-point of the study, the 12-month reduction in LVEDD and LVESD was greater in idiopathic DCM (by 8.9 and 9.8mm, P<0.01, respectively) compared with ischaemic cardiomyopathy. The 6-min walk rose by 15% (P<0.01) and peak VO(2)by 10% (P<0.05) after 3 months, with no further increase by 12 months, and no difference between idiopathic DCM and ischaemic cardiomyopathy. The increase in peak VO(2)at 12 months correlated with the fall in ESD (r=0.47, P=0.03). CONCLUSIONS: A-BiV pacing shortens total IVT, reduces LV cavity size, and increases exercise tolerance in patients with DCM and IVCD. Ischaemic cardiomyopathy does not affect the exercise response, although it does reduce the extent of reverse remodelling. PMID- 12633546 TI - The EuroHeart Failure survey programme-- a survey on the quality of care among patients with heart failure in Europe. Part 1: patient characteristics and diagnosis. AB - BACKGROUND: The European Society of Cardiology (ESC) has published guidelines for the investigation of patients with suspected heart failure and, if the diagnosis is proven, their subsequent management. Hospitalisation provides a key point of care at which time diagnosis and treatment may be refined to improve outcome for a group of patients with a high morbidity and mortality. However, little international data exists to describe the features and management of such patients. Accordingly, the EuroHeart Failure survey was conducted to ascertain if appropriate tests were being performed with which to confirm or refute a diagnosis of heart failure and how this influenced subsequent management. METHODS: The survey screened consecutive deaths and discharges during 2000-2001 predominantly from medical wards over a 6-week period in 115 hospitals from 24 countries belonging to the ESC, to identify patients with known or suspected heart failure. RESULTS: A total of 46788 deaths and discharges were screened from which 11327 (24%) patients were enrolled with suspected or confirmed heart failure. Forty-seven percent of those enrolled were women. Fifty-one percent of women and 30% of men were aged >75 years. Eighty-three percent of patients had a diagnosis of heart failure made on or prior to the index admission. Heart failure was the principal reason for admission in 40%. The great majority of patients (>90%) had had an ECG, chest X-ray, haemoglobin and electrolytes measured as recommended in ESC guidelines, but only 66% had ever had an echocardiogram. Left ventricular ejection fraction had been measured in 57% of men and 41% of women, usually by echocardiography (84%) and was <40% in 51% of men but only in 28% of women. Forty-five percent of women and 22% of men were reported to have normal left ventricular systolic function by qualitative echocardiographic assessment. A substantial proportion of patients had alternative explanations for heart failure other than left ventricular systolic or diastolic dysfunction, including valve disease. Within 12 weeks of discharge, 24% of patients had been readmitted. A total of 1408 of 10434 (13.5%) patients died between admission and 12 weeks follow-up. CONCLUSIONS: Known or suspected heart failure comprises a large proportion of admissions to medical wards and such patients are at high risk of early readmission and death. Many of the basic investigations recommended by the ESC were usually carried out, although it is not clear whether this was by design or part of a general routine for all patients being admitted regardless of diagnosis. The investigation most specific for patients with suspected heart failure (echocardiography) was performed less frequently, suggesting that the diagnosis of heart failure is still relatively neglected. Most men but a minority of women who underwent investigation of cardiac function had evidence of moderate or severe left ventricular dysfunction, the main target of current advances in the treatment of heart failure. Considerable diagnostic uncertainty remains for many patients with suspected heart failure, even after echocardiography, which must be resolved in order to target existing and new therapies and services effectively. PMID- 12633547 TI - The EuroHeart Failure Survey programme--a survey on the quality of care among patients with heart failure in Europe. Part 2: treatment. AB - BACKGROUND: National surveys suggest that treatment of heart failure in daily practice differs from guidelines and is characterized by underuse of recommended medications. Accordingly, the Euro Heart Failure Survey was conducted to ascertain how patients hospitalized for heart failure are managed in Europe and if national variations occur in the treatment of this condition. METHODS: The survey screened discharge summaries of 11304 patients over a 6-week period in 115 hospitals from 24 countries belonging to the ESC to study their medical treatment. RESULTS: Diuretics (mainly loop diuretics) were prescribed in 86.9% followed by ACE inhibitors (61.8%), beta-blockers (36.9%), cardiac glycosides (35.7%), nitrates (32.1%), calcium channel blockers (21.2%) and spironolactone (20.5%). 44.6% of the population used four or more different drugs. Only 17.2% were under the combination of diuretic, ACE inhibitors and beta-blockers. Important local variations were found in the rate of prescription of ACE inhibitors and particularly beta-blockers. Daily dosage of ACE inhibitors and particularly of beta-blockers was on average below the recommended target dose. Modelling-analysis of the prescription of treatments indicated that the aetiology of heart failure, age, co-morbid factors and type of hospital ward influenced the rate of prescription. Age <70 years, male gender and ischaemic aetiology were associated with an increased odds ratio for receiving an ACE inhibitor. Prescription of ACE inhibitors was also greater in diabetic patients and in patients with low ejection fraction (<40%) and lower in patients with renal dysfunction. The odds ratio for receiving a beta-blocker was reduced in patients >70 years, in patients with respiratory disease and increased in cardiology wards, in ischaemic heart failure and in male subjects. Prescription of cardiac glycosides was significantly increased in patients with supraventricular tachycardia/atrial fibrillation. Finally, the rate of prescription of antithrombotic agents was increased in the presence of supraventricular arrhythmia, ischaemic heart disease, male subjects but was decreased in patients over 70. CONCLUSION: Our results suggest that the prescription of recommended medications including ACE inhibitors and beta-blockers remains limited and that the daily dosage remains low, particularly for beta-blockers. The survey also identifies several important factors including age, gender, type of hospital ward, co morbid factors which influence the prescription of heart failure medication at discharge. PMID- 12633548 TI - Effects of a perindopril-based blood pressure lowering regimen on cardiac outcomes among patients with cerebrovascular disease. AB - AIM: To determine the effects of a perindopril-based blood pressure lowering regimen on major cardiac events among hypertensive and non-hypertensive patients with a history of cerebrovascular disease. METHODS AND RESULTS: A total of 6105 individuals with a history of stroke or transient ischaemic attack were randomly assigned active treatment (n=3051) or placebo (n=3054). Active treatment comprised the angiotensin-converting-enzyme inhibitor perindopril (4 mg daily), with the addition of the diuretic indapamide at the discretion of treating physicians. Over a mean of 3.9 years of follow-up, active treatment reduced blood pressure by 9/4 mm Hg compared with placebo and reduced the primary outcome, stroke, by 28%. Major coronary events occurred in 269 participants (active 3.8%, placebo 5.0%) and heart failure was diagnosed in 264 participants (active 3.7%, placebo 4.9%). Active treatment reduced the risk of major coronary events by 26% (95% CI: 6-42%; p=0.02) and the risk of congestive heart failure by 26% (5-42%; p=0.02). For each of these outcomes, there was no clear evidence of differences between the treatment effects in participants classified as hypertensive or non hypertensive, and those with or without a history of coronary heart disease. CONCLUSIONS: Among individuals with cerebrovascular disease, blood pressure lowering with a regimen involving perindopril and indapamide not only reduced the risk of stroke, but also substantially reduced the risks of cardiac outcomes. PMID- 12633549 TI - Axillary recurrence following sentinel node biopsy for breast cancer. PMID- 12633550 TI - Towards a scientific basis for oncoplastic breast surgery. PMID- 12633551 TI - APC, beta-Catenin and hTCF-4; an unholy trinity in the genesis of colorectal cancer. AB - Mutations in APC have been identified in up to 80% of 'classic' sporadic colorectal cancers. Although the APC gene was first sequenced over a decade ago, new functions are still being described and its importance in the genesis of colorectal cancer continues to increase. The current focus of attention is on the APC/beta-Catenin/TCF signal transduction pathway as the main effector mechanism, and recent work has also implicated this pathway in the aetiology of the minority of CRCs that develop through mismatch repair. At the same time, new evidence on the interactions of APC with the cytoskeleton and the demonstration of a nuclear export function in the protein have shown that it has multiple additional roles in colorectal carcinogenesis. Thus this is an area that benefits from further review of the ever expanding literature. PMID- 12633552 TI - Sentinel lymph node biopsy in breast cancer after neoadjuvant chemotherapy. A pilot study. AB - AIMS: Sentinel lymph node biopsy (SLN) seems to represent a reliable method for early breast cancer staging, offering an alternative to complete axillary dissection. METHODS: The identification of sentinel node(s), their localization and the predictive capacity of SLN were analysed in 42 patients who had neoadjuvant chemotherapy for breast cancer. Dextran labelled with (99m)Tc was injected close to the tumor. The radioactive uptake in the axilla was detected by scintigraphic images and by a hand-held probe guided the SLN. RESULTS: One or more sentinel nodes were identified in 41 patients (97.6%). We could accurately predict axillary lymph nodes status in 93% of the cases. CONCLUSIONS: SLN can predict the status of the axilla in patients who have received primary chemotherapy. PMID- 12633553 TI - Antigenic profiles of disseminated breast tumour cells and microenvironment in bone marrow. AB - AIMS: Thirty per cent of breast cancer patients with axillary lymph node negative at primary surgery will relapse within 10 years. This may be caused by disseminated tumour cells from the primary tumour. This study report the phenotypic profiles of disseminated tumour cells and microenvironmental characteristics in bone marrow of breast cancer. METHODS: We detected the biologic markers on the disseminated tumour cells with immunocytochemical staining, analysed the immunological changes through flow-cytometry, and investigated the u- PA activity in the plasma of bone marrow. RESULTS: With the immunocytochemical staining of EMA and CK19, we detected micrometastasis in thirty out of 72 (41.67%) breast cancer patients. Compared with the primary tumours, disseminated tumour cells expressed low protein cyclin D1, P53, Ki-67, EGFR, and high protein P21. The percentage of memory CD4+ T cells was significantly higher in the micrometastasis-positive group than in the micrometastasis-negative group. Tumour size and axillary lymph node status were found to be significantly correlated with the u- PA activity level. CONCLUSIONS: Immunophenotypic profiles of disseminated tumour cells could be measured by immunocytochemical staining and microenvironment can be analysed by flow cytometry. PMID- 12633554 TI - Endoscopic muscular latissimus dorsi flap harvesting for immediate breast reconstruction after skin sparing mastectomy. AB - AIM: Immediate breast reconstruction after mastectomy using the latissimus dorsi musculo-cutaneous flap is well recognized. It allows for satisfactory aesthetic results. To minimize scanning in skin sparing mastectomy patients, we used a surgical technique consisting of an endoscopic harvesting of the latissimus dorsi pure muscular flap with a virtual cavity created by CO(2) gas distention. METHOD: Between 9 April 2001 and 30 September 2001, 8 patients underwent latissimus dorsi endoscopic harvesting for an immediate breast reconstruction after skin sparing mastectomy. RESULT: The mean operating endoscopic time was 112 minutes. No open surgical conversion was necessary. The mean lymphatic drainage was 2720 ml with removal of the drainage on post operative day 15. CONCLUSION: The endoscopic harvesting of the latissimus dorsi muscular flap, using a mixed technique of dissection guided by the lighted cleaver forceps and a closed technique by CO(2) insufflation using an endoscope is feasible, reproducible and has acceptable morbidity. This technique brings encouraging aesthetic results after skin sparing mastectomy. PMID- 12633555 TI - The prognostic significance of total lymph node number in patients with axillary lymph node-negative breast cancer. AB - AIM: In node-negative breast cancer patients, several factors for survival have been evaluated and currently, some of them are accepted for their prognostic and/or predictive values after validation in the separate data sets. The prognostic significance of increases in the number of pathologically detectable axillary lymph nodes in the node-negative patients could not been established clearly. To address this question, we have reviewed our patients' records. METHODS: A retrospective cohort study was conducted in pathologically node negative patients who underwent modified radical mastectomy for stage I and II breast cancer. Survival and multivariate prognostic factor analyses were carried out to determine whether the number of tumour-free lymph nodes in complete axillary dissection material in addition to known factors was significant for the outcomes. RESULTS: Two hundred and seventy consecutive patients were eligible to enter the trial. The median observation time and the median number of tumour-free lymph nodes were 61 (from 30 to 120) months and 18 (from 10 to 44), respectively. The cohort was divided into the groups according to the number of nodes. The 5 year event-free and overall survivals were 92.5 and 98.3% for patients who had 18 lymph nodes or less, and 70 and 86.7% for those who had more than 18 negative nodes, respectively (P < 0.00001). Multivariate analysis for event-free survival demonstrated that the number of lymph nodes (Relative risk: 3.2 and 95% confidence interval: 1.7 to 5.9) in addition to the pathological tumour size and age was the most important independent prognosticator. In similar, multivariate analysis for overall survival showed that the number of lymph nodes together with the tumour size was the significant indicator (RR of cancer-specific dying in patients who had more than 18 nodes: 3.1 and 95% CI: 1.2 to 8.5). CONCLUSION: The increases in number of tumour-free lymph nodes are clinically important and this parameter should be taken into consideration in the breast cancer patients without metastatic lymph nodes. PMID- 12633556 TI - Surgeon-controlled ultrasound-guided core biopsies in the breast--a prospective study and a new use for surgeons in the clinic. AB - AIMS: This study was to assess the accuracy of a surgeon performing ultrasound guided core biopsies of the breast. METHODS: A prospective audit was carried out of 555 patients who underwent an ultrasound-guided core biopsy for a discrete solid mass [under 30 mm maximum diameter] by a single surgeon. The surgeon controlled the core biopsy needle and an ultrasonographer or radiologist provided the imaging with ultrasound. RESULTS: The accuracy of the surgeon in sampling the lesions [n = 555 core biopsies] was independent of the size of the lesion. This saved 272 patients having unnecessary surgery for a benign lesion. CONCLUSION: The practical involvement by the surgeon in breast ultrasound and performing core biopsies has reduced pressure on the breast radiologists, reduced the number of diagnostic surgical open biopsies and made the clinic more interesting for the surgeon. With increased surgical confidence in breast ultrasound, most small and impalpable cancers have pre-operative skin marking rather than X-ray wire localization prior to undergoing wide local excision. Trainee breast surgeons should be encouraged to learn breast ultrasound and core biopsies. PMID- 12633557 TI - Uncontrolled local disease after salvage treatment for ipsilateral breast tumour recurrence. AB - AIM: Uncontrolled local disease (ULD) following breast conservation constitutes a clinical problem with a major impact on quality of life. The current study analysed the outcome following treatment of ipsilateral breast tumour recurrence (IBTR) and the risk for ULD with the aim to identify risk factors for ULD. METHODS: In a cohort of 5502 patients treated for invasive breast cancer Stage I II with breast-conserving surgery 1976-1998 in Stockholm, 307 patients with subsequent IBTR were identified. The majority (n = 219) had received postoperative radiotherapy. Twenty-six per cent of the patients received adjuvant tamoxifen, for 2 or 5 years, and 9% received adjuvant polychemotherapy. Median follow-up time was 11(2-23) years. 50/307 patients developed ULD, defined as the appearance of clinically manifest invasive adenocarcinoma in the remaining breast or on the ipsilateral chest wall which could not be eradicated within 3 months of detection. Multivariate linear logistic regression was used in the statistical analysis to identify prognostic factors for ULD. RESULTS: Five years following the diagnosis of IBTR the cumulative incidence of ULD was 13%. Five independent risk factors for ULD were identified; non-surgical treatment of IBTR, disseminated disease concurrent with IBTR, axillary lymph node metastases (at primary breast conservation), time < 3 years between breast conservation and IBTR, no adjuvant endocrine therapy. Eighty-eight per cent of the patients were treated with salvage mastectomy (n = 207) or re-excision (n = 62). The cumulative incidence at 5 years of ULD following salvage mastectomy and salvage re-excision were 10% and 16% respectively compared to 32% among patients treated non surgically. Following IBTR, the 5-year overall survival among patients with local control was 78% in contrast to 21% among patients with ULD. CONCLUSION: Uncontrolled local disease is an infrequent but important outcome following breast-conserving surgery. Primary postoperative radiotherapy reduces the risk for IBTR and is therefore recommended as part of the primary treatment to avoid both IBTR and ULD. In addition to radiotherapy, adjuvant therapy reduces the risk for IBTR and thereby the risk for subsequent ULD. Patients with IBTR, independent of concurrent distant metastases, should when feasible be recommended for salvage surgery as it provides superior local control compared to salvage systemic therapy alone. PMID- 12633558 TI - Impaired postoperative neutrophil leucocytosis and acute complications following short course preoperative radiotherapy for operable rectal cancer. AB - AIM: Postoperative mortality following total mesorectal excision is increased if this is performed more than 3 days after the completion of short course preoperative radiotherapy. Suppression of neutrophil leucocytosis which is normally seen following surgery has been a suggested reason. This study was to determine the relationship between postoperative complications and perioperative neutrophil counts. METHOD: A database of 176 patients treated at a single radiotherapy centre in 1998 and 1999 was used. A two-sample Wilcoxon test was used to compare preoperative neutrophil counts, postoperative neutrophil counts and their ratio in patients with and without complications. RESULTS: There was no association between acute complications and preoperative (P = 0.25) or postoperative (P = 0.45) neutrophil count. The post/preoperative ratio was significantly higher in patients without complications (median 1.61 vs 1.16, P = 0.02). CONCLUSIONS: There appears to be an inverse relationship between the magnitude of postoperative neutrophil leucocytosis and the risk of acute complications following short course preoperative radiotherapy. PMID- 12633559 TI - Surgery after induction chemoradiotherapy for oesophageal cancer. AB - AIM: We discuss the role of surgery after induction chemoradiotherapy, when a complete response is apparent. METHODS: We enrolled 247 consecutive patients who underwent chemoradiotherapy and oesophagectomy for cancer. Patients were classified in two groups as to whether they had chemoradiotherapy (n = 60) or not (n = 187) before surgery. RESULTS: The 5-year survival rate of patients with complete response was 54% and was significantly higher than that of the other group (P = 0.018). Of the 60 patients responding, 34 (56.7%) were found to have a complete pathological tumour response (pT0). The overall sensitivity and specificity of the post-chemoradiotherapy restaging were 60.7% and 86.4%. CONCLUSIONS: Complete response after induction chemoradiotherapy is not a reliable tool in the management of oesophageal cancer. We need to improve postchemoradiotherapy restaging to be sure that chemoradiotherapy will provide the same long-term survival rates as surgery. PMID- 12633560 TI - Preoperative radiotherapy in combination with metronidazole for resectable gastric cancer: long-term results of a phase 2 study. AB - AIM: Surgery remains the main treatment of stomach carcinoma. Poor long-term survival mandates the study of other techniques. An original treatment scheme with preoperative radiotherapy and metronidazole as a radiosensitizer is reported here. METHODS: Between 1982 and 1988, MRRC RAMS carried out a prospective clinical trial of preoperative radiotherapy (20 Gy/5 days) in combination with metronidazole (given orally, 3 times, 5 gms per metre(2)). Of 91 patients who received preoperative radiotherapy, 67 patients were operated on with curative intent and were eligible for further analysis. RESULTS: Acute gastro-intestinal toxicity was significant but manageable without surgery delay in most cases. There were 4 postoperative deaths. Overall 5-year, 10-year survival and median survival were 46%, 36% and 46 months. Serosal and nodal involvement were the most significant adverse prognostic factors. Tumours confined to the gastric wall, node negative cases, middle and distal location, differentiated tumours, female sex and age more than 50 years were associated with relatively good long-term results: overall 5-year survival was 50% or better, overall 10-year survival was 40% or better. CONCLUSION: The combined treatment showed significant but manageable acute toxicity. The long-term results seem encouraging and support further investigations in multimodal treatment of gastric cancer. PMID- 12633561 TI - Cyclooxygenase-2 in human non-small cell lung cancer. AB - AIM: Recent studies report that the expression of cyclooxygenase (COX) in non small cell lung cancer (NSCLC) is increased, especially in adenocarcinoma. Platelet activating factor (PAF), n-sodium butyrate (n-BT), and phorbol myristate acetate (PMA) are important mediators of the inflammatory process. METHOD: Expression of COX-2 in 67 stage 1 NSCLC paraffin-embedded tumor samples was determined by immunohistochemistry (IHC). Four NSCL cell lines were incubated and stimulated by PAF, n-BT and PMA for 48 h. Expression of COX-2 was determined by IHC, immunoblotting, and reverse transcription-polymerase chain reaction (RT PCR). RESULT: IHC showed increasing immunoreactivity in 35 of 67 (52%) in stage I NSCLC, 31 of 53 (59%) in adenocarcinoma and 13 of 15 (87%) in bronchoalveolar cell carcinoma, but only 2 of 12 (17%) in epidermoid carcinoma. The COX-2 expression in NSCLC cells was 75% (3/4) and the COX-1 expression in NSCLC cells was 100% (4/4). After stimulation with PMA, n-BT, PAF and n-BT + PAF, the COX-2 expression in NSCLC cells was significantly increased in all cell lines. CONCLUSIONS: The expression of COX-2 in NSCLC cells is high and was up-regulated by PMA, n-BT and PAF. We consider that COX-2 inhibitors will play an important role in the therapy of NSCLC. PMID- 12633562 TI - Heterogeneity of stage IIIA non-small cell lung cancers (NSCLC) and evaluation of late results of surgical treatment. AB - AIMS: The aim of the study was assessment of the heterogeneity of stage IIIA non small cell lung cancers (NSCLC) and the late results of surgical treatment. METHODS: The study group consisted of 83 consecutive patients discharged between 1988 and 1992 undergoing radical operative treatment for stage IIIA NSCLC. Squamous cell carcinoma was diagnosed in 54 (65.1%) patients, adenocarcinoma in 23 (27.7%), large cell carcinoma in 2 (2.4%) and mixed (i.e. adenoid-squamous type) in 4 (4.8%). In respect of pTNM staging, 19 (22.9%) patients had T3N1M0, 35 (42.2%) had T2N2M0 and 29 (34.9%) had T3N2M0. RESULTS: Overall, 13.3% of patients with stage IIIA NSCLC survived 5 years following the operation and 8.7% survived 10 years. Analysis of follow-up study indicated that this group was heterogenic. In T3N1M0 group 26.3% survived 5 years following the operation, in T2N2M0 group 14.3%, in T3N2M0 group 3.5% (P = 0.015). Of 23 patients with N2 disease and no metastases in hilar lymph nodes ('skip' metastases), 26.1% survived 5 years, whereas none of 41 patients with metastases spreading by continuity survived (P = 0.0015). If mediastinal lymph node metastases were diagnosed in one level, 25% patients survived 5 years, but if two or more levels were affected, 2.3% only (P = 0.0214): 85.7% of patients with well-differentiated (G1) cancer survived 5 years and 62.0% 10 years, whereas among those with moderately differentiated (G2) tumours, 11.8% and 8.8%, respectively. No patient survived 5 years after resection of poorly differentiated (G3) cancer (P < 0.001). CONCLUSIONS: (1) Patients operated for stage IIIA NSCLC are a heterogeneous group, which makes it difficult to predict late results. (2) Patients operated for stage IIIA NSCLC have a better prognosis if metastases are discovered in level one mediastinal lymph nodes, particularly in the superior part of mediastinum, or if 'skip' metastases (pulmonary hilus unaffected) are discovered, as compared to those with N2 disease. (3) Poor histologic differentiation of the tumour is a bad prognostic factor. PMID- 12633564 TI - Minimally invasive video assisted parathyroidectomy (MIVAP). AB - The first endoscopic approach to parathyroid glands was reported by M. Gagner in 1996. Later, different accesses have been described using either CO(2) insufflation or external retraction. Other widespread procedures include the lateral access proposed by J.F. Henry and the central gas-less access proposed by P. Miccoli. We hereby describe this central access which allows a bilateral exploration of the neck when necessary. Our patient data base consists of 270 patients operated on since February 1997. The mean age was 56.5 years (20-87 years). The female to male ratio was 4:1. The mean operative time of the procedure was 41.1 min (range 15-180 min). In 13 cases, a video-associated thyroid resection was accomplished during the same operation for associated diseases. Conversion to traditional cervicotomy was required in 20 patients (8.09%). One laryngeal nerve palsy was confirmed 6 months after surgery. We registered one postoperative bleeding, which required us to reoperate on the patient 2 hours after first surgery. The mean operative time and complication rate clearly demonstrate that this approach, like other minimally invasive techniques, can successfully rival the results of traditional surgery for the treatment of primary hyperparathyroidism. PMID- 12633563 TI - A report of six cases of familial papillary thyroid cancer. AB - AIMS: Familial occurrence of papillary thyroid cancer is uncommon. The purpose of this study was review our own experience in a series of 267 papillary thyroid cancers. METHODS: We analysed the clinical records of 267 consecutive patients operated on for papillary thyroid cancer (PTC) in our hospital between June 1980 March 2000. RESULTS: We identified a family history in three families (6 patients), which results in a 2.25% familial papillary thyroid carcinoma (FPTC) rate. Pathology findings revealed that the tumour was multifocal and bilateral in 2 patients. Lymph-node metastases were found in 4 patients. They are all alive with a mean time of follow-up of 74.3 months (range 2-120). CONCLUSIONS: We recommend that patients with familial disease should be treated according to the disease stage and other risk factors, similar to those with sporadic differentiated papillary thyroid cancer. We encourage the further reporting and pedigree analysis to identify patients affected by FPTC. PMID- 12633566 TI - Release of cytokines and apoptosis in fetal rat Type II pneumocytes exposed to hyperoxia and nitric oxide: modulatory effects of dexamethasone and pentoxifylline. AB - The response of the fetal rat Type II pneumocyte (FTIIP), the stem cell of the alveolar epithelium, to hyperoxia would be helpful to understand the effects of oxygen-induced injury to the immature lung. In such a scenario, the presence of nitric oxide (NO) may have a protective or detrimental effect. Our goals were to evaluate the release of cytokines and apoptotic cell death in freshly isolated FTIIP (19-day) in the presence of 95% O(2) and/or NO. The effects of dexamethasone and pentoxifylline on the FTIIP cytokine response were also studied. There was no significant difference in the levels of IL-1beta and IL-10 from FTIIP, in room air, hyperoxia and/or NO at 2, 6 and 24 h. However, IL-6 release was significantly higher, when measured over time, after 2, 6 and 24 h of exposure to hyperoxia and NO. Dexamethasone in the presence of hyperoxia and/or NO increased the release of IL-10 at 24 h. There was increased apoptosis in FTIIP exposed to hyperoxia alone and in combination with NO; this was significantly attenuated in the presence of dexamethasone and pentoxifylline. We speculate that the cytoprotective effects of dexamethasone in the immature lung may, in part, be mediated via IL-10. PMID- 12633567 TI - Effects of interleukin-8 on the developing human intestine. AB - The human fetal/neonatal gastrointestinal tract is exposed to biologically significant concentrations of interleukin (IL)-8 swallowed with amniotic fluid and human milk. We hypothesized that IL-8 has a physiologic function in the developing human intestine. IL-8 was measured in preterm and term human milk, tested for stability under conditions simulating neonatal gastric and proximal small intestinal digestion, and its receptors were sought in human fetal bowel. The effect of IL-8 was then measured on intestinal cells in vitro. We observed that IL-8 is present in significant concentrations in human milk and that it is stable under conditions simulating digestion. Both IL-8 receptors, CXCR1 and CXCR2, are expressed extensively in the fetal intestine. When human fetal and adult intestinal cells are treated with rhIL-8 in vitro, there is a consistent increase in cell migration, proliferation, and differentiation. IL-8 also protects intestinal cells against chemical injury. These results suggest that besides its better-known role as a neutrophil chemoattractant, IL-8 has a trophic function in the developing human intestine. PMID- 12633568 TI - Increased expression of Vascular Endothelial Growth Factor-A in seasonal allergic rhinitis. AB - Increased vascular dilatation and permeability characterize allergic rhinitis. In this study oligonucleotide microarrays (Affymetrix HuGe95A) were used to identify differentially expressed vasoactive genes in nasal biopsies from 23 patients with symptomatic seasonal allergic rhinitis (SAR) and 12 healthy controls. RNA was extracted from the biopsies and pooled in three patient and three control pools. Out of 12,626 analysed transcripts, 39 were higher and 81 lower in the patients. Of these transcripts two have vasoactive effects: Vascular Endothelial Growth Factor-A (VEGF-A) and the Beta-1-Adrenergic Receptor. Both were higher in patients than in controls. The mean +/- SEM expression levels in arbitrary units of VEGF-A were 130 +/- 123 in the patients and 59 +/- 53 in the controls. The fold ratio in expression levels between patients/controls was 2.2. The corresponding values for the beta-1-adrenergic receptor were 129 +/- 123 in the patients and 40 +/- 31 in the controls. The fold ratio between patient/controls was 3.2. The role of VEGF-A was assessed by determining VEGF-A concentrations in nasal fluids from another 30 patients with SAR before and after allergen provocation. VEGF-A increased from 124.3 +/- 30.2 to 163.2 +/- 37.8 pg/ml after challenge, P < 0.05. In summary, oligonucleotide microarray analysis of nasal biopsies and protein analyses of nasal fluids indicate that VEGF-A may be an important mediator in SAR. PMID- 12633565 TI - Towards optimal management of ductal carcinoma in situ of the breast. AB - Ductal carcinoma in situ (DCIS) represents a spectrum of heterogenous disease that accounts for approximately one fifth of all screen-detected breast cancers and is considered as a precursor of invasive breast cancer if left untreated (35 50% risk). DCIS can be treated by total mastectomy with or without immediate breast reconstruction, local excision (LE) plus adjuvant radiotherapy (RT) or LE alone. Total mastectomy is associated with low rates of local recurrence (1.4%) and breast cancer-specific mortality (0.59%). Three recent randomized controlled trials (RCTs) have demonstrated that adjuvant RT after LE of localized DCIS significantly reduces the incidence of local recurrence. However these trials did not identify any subgroups of patients where RT could be safely omitted. Retrospective studies suggest that RT can be safely omitted after adequate LE (margin width > or =1 cm) of small (< 15 mm), non-high grade DCIS not associated with necrosis. Further RCTs are required to validate these retrospective findings, with an emphasis on standardized and meticulous tissue processing and pathological evaluation. The role of adjuvant tamoxifen in the management of DCIS continues to evolve. Formal axillary dissection is not appropriate for DCIS, however, the potential role of the sentinel node biopsy (SNB) in selected high risk cases requires further evaluation. The International Breast Cancer Intervention Study (IBIS-II) trial aims to evaluate the potential role of third generation aromatase inhibitors in postmenopausal women with hormone-sensitive DCIS.Future research will focus on the relevance of gene expression profiling, proteomics, Laser therapy and mammary ductoscopy to the management of DCIS. PMID- 12633569 TI - Cloning, sequencing and expression of porcine CD40 ligand in Escherichia coli and human and porcine cells. AB - The CD40L ligand (CD40L) plays an important role in the interaction between antigen-specific T lymphocytes and antigen-presenting cells. The porcine CD40L encoding gene was isolated from porcine peripheral blood mononuclear cells (PBMC) using RT-PCR. Sequence analysis of the cloned CD40L gene showed an open reading frame of 786 base pairs encoding a 262 amino acid protein with a predicted molecular mass of 29 kD. The deduced amino acid sequence of the porcine CD40L shared 82%, 88% and 93% similarity with the CD40L protein of mouse, human and cattle. The isolated CD40L sequence was expressed as a hexahistidine fusion protein in Escherichia coli and purified by affinity chromatography. The analysis of the CD40L-expression in human 293 and porcine MAX cells by immunofluorescence showed its location on the cell surface. PMID- 12633570 TI - Potentiation of Fas- and TRAIL-mediated apoptosis by IFN-gamma in A549 lung epithelial cells: enhancement of caspase-8 expression through IFN-response element. AB - Epithelial cell apoptosis triggered cooperatively by multiple cytokines contributes to the injury induced by inflammatory responses in the lung and elsewhere. Here we show that interferon-gamma (IFN-gamma) sensitizes A549 cells, human lung epithelial cells, to cytokine-mediated apoptosis by upregulating caspase-8 expression. Pretreating the cells with IFN-gamma potentiated Fas- and TNF-related apoptosis inducing ligand (TRAIL)-induced cell death, but other forms of apoptosis, not mediated via receptors, were unaffected. Western blotting and inhibitor assays showed that IFN-gamma selectively increased expression of caspases-7 and -8, but not caspases-2, -3, -9, or -10, as a necessary step leading to apoptosis. Assaying promoter activity using a luciferase reporter gene indicated that an IFN-gamma response element was located in the 5'-flanking region of the caspase-8 gene, spanning positions -227 to -219. Taken together, these findings suggest that IFN-gamma potentiates Fas- and TRAIL-mediated apoptosis by increasing caspase-8 expression via an IFN-gamma response element in A549 cells. PMID- 12633571 TI - Enhanced transcytosis of R5-tropic human immunodeficiency virus across tight monolayer of polarized human endometrial cells under pro-inflammatory conditions. AB - Most cases of human immunodeficiency virus (HIV) infection worldwide occur following sexual contact, implying that the virus may breach the protective epithelial barrier lining the genital tract. HIV infection is known to preferentially occur when the genital epithelial integrity is altered, particularly when epithelial micro-ulcerations occur during heterosexual intercourse or ulcerations appear, due to sexually transmitted infections or else in the context of ectopy of the endocervical mucosa, which may leave the genital tissue. We report that R5-tropic infectious HIV-1 isolates are capable of in vitro transcytosis through a tight and polarized monolayer of human endometrial HEC-1 cells. Transcytosis of HIV particles was increased 2-fold within a pro inflammatory micro-environment. Our findings suggest that transcytosis may be a relevant mechanism for the passage of virus through the genital mucosa in vivo, particularly when inflammatory cells and mediators are present in the vicinity of the mucosal surface. PMID- 12633572 TI - Contribution of circulating leukocytes to cytokine production in pancreatic duct obstruction-induced acute pancreatitis in rats. AB - Little information is available regarding the role of circulating leukocytes in the pathogenesis of acute pancreatitis (AP). Our aim was to explore the time course of the potential role of inflammatory peripheral blood (PB) cells during AP induced in rats by pancreatic duct obstruction (PDO). Flow cytometry immunophenotyping was used to analyse the distribution of the major circulating leukocyte subsets, the activation state of circulating monocytes as reflected by both CD11b expression and TNF-alpha production and the relative contribution of T cell derived pro- (TNF-alpha) and anti- (IL-10) inflammatory mediators at different stages of PDO-induced AP. A progressive increase in PB neutrophils and monocytes was observed up to 6h after PDO whereas lymphocytes, as well as CD4(+) and CD8(+) T-cell subsets, rose as early as 1.5 h after PDO and decreased thereafter. Monocytes were activated in PB from 6 h after inducing AP as reflected by increases in both CD11b expression and spontaneous TNF-alpha production; nevertheless, they showed the capability of producing TNF-alpha at earlier AP stages by lipopolysaccharide (LPS) stimulation. In contrast, T-cells were unable to produce TNF-alpha during AP neither spontaneously nor after stimulation with PMA/Ionomycin. Therefore, only PB monocytes contribute to increase TNF-alpha levels in plasma as observed from 12 h onwards after inducing AP. Interleukin-10 was produced by T-cells 6 h after PDO only after PMA/Ionomycin stimulation. We conclude that systemic inflammatory events are triggered off at early stages of PDO-induced AP, with the activation of circulating monocytes, though not T-cells, playing a central role. PMID- 12633573 TI - Anti-interleukin 6 (IL-6) receptor antibody suppresses Castleman's disease like symptoms emerged in IL-6 transgenic mice. AB - Transgenic mice carrying human IL-6 cDNA fused with a murine major histocompatibility class-I promoter (H-2L(d)) were serially administered with anti-interleukin-6 receptor (IL-6R) monoclonal antibody (mAb), MR16-1, from the age of 4 weeks to estimate its efficacy on a variety of disorders developed in these mice, most of which are similar to the disorders associated with Castleman's disease. In the control mice treated with isotype-matched mAb, a massive and multiple IgG1 plasmacytosis, mesangial proliferative glomerulonephritis, leukocytosis, thrombocytosis, anemia and abnormalities of blood chemical parameters have developed in accordance with the elevation of serum IL-6, and 50% of mice have died of renal failure by 18 weeks of age. In contrast, the treatment with MR16-1 prevented all these symptoms and prolonged the lifetime of the majority of the mice. Thus, the constitutive overexpression of IL-6 caused various disorders, and the treatment with anti-IL-6R mAb completely prevented from these symptoms. These results clearly confirm that IL-6 indeed plays an essential role in the pathogenesis of a variety of disorders. Furthermore, anti-IL-6R mAb could provide novel therapy for Castleman's disease and MR16-1 should be a useful tool to estimate therapeutic potential of IL-6 antagonists in a variety of murine models for human disease. PMID- 12633575 TI - Haemopoietic stem cell transplantation for acute lymphoblastic leukaemia. AB - The majority of children and some adults with acute lymphocytic leukaemia (ALL) can be cured with current intensive chemotherapy regimens. For those patients who relapse or who do not achieve remission, allogeneic haemopoietic stem cell transplantation (HSCT) offers the best chance for long-term disease control. Different sources of haemopoietic stem cells including marrow, peripheral blood, and cord blood are now available and the introduction of subablative regimens has increased the number of patients who are transplant candidates. Relapse remains the major cause of transplant failure and immunotherapy strategies post transplant to augment the graft versus leukaemia effect are being explored. PMID- 12633576 TI - Influence of differing radiotherapy strategies on treatment results in diffuse large-cell lymphoma: a review. AB - In the absence of evidence from randomised trials, radiation treatment of diffuse large-cell lymphoma of B-cell type represents an area of controversy with considerable differences in patterns of practice. The present literature survey aims at clarification of the role of radiotherapy in combined modality settings by identification of dose-response relationships, predictive factors for local control, and potential pitfalls in the interpretation of retrospective studies. Radiotherapy might increase local control in initially involved areas and is usually delivered to these sites (involved-field treatment). Combined modality treatment is currently recommended for patients in stage I or II if they are not treated in the context of prospective studies. Whether involved-field consolidation radiotherapy after systemic treatment in patients with bulky, stage III-IV lymphomas should be routinely recommended is presently unclear. Definition of bulky disease is arbitrary and varied between 6 and 10cm, reflecting a considerable difference in the number of clonogenic tumour cells. Several retrospective and one prospective randomised study suggest improved disease-free and overall survival by radiotherapy in advanced stages. The 5-year local control by radiotherapy was 93-98%. Currently, we recommend the following minimum doses for involved-field radiotherapy derived from this literature survey. Lymphomas with initial size <3.5 cm (possibly <6 cm) can be treated with 30 or 30.6Gy when a complete remission (CR) has been achieved by chemotherapy. The next group might be sufficiently controlled by 36Gy, but it remains unclear whether the cut-off should be 6cm or higher. Forty Gy appears to control tumours in the range of 7 10cm. Most likely, 45Gy does not have to be exceeded for larger lesions. Data on those with less than CR are contradictory. Judging the amount of viable tumour in these patients is problematic, but crucial to determine the intensity of further treatment. The value of positron emission tomography is still under investigation. Because the difference between doses of 30 and 40Gy might actually make a difference for the long-term toxicity of radiotherapy in some of the normal tissues and organs at risk (salivary glands, orbital structures, lung, heart, etc.), it appears prudent to resolve the open questions in prospective trials with careful documentation of side effects. PMID- 12633578 TI - Recent advances in the treatment and understanding of childhood acute lymphoblastic leukaemia. AB - Clinical trials have advanced the cure rate of childhood acute lymphoblastic leukaemia to near 80%. Treatment response, as measured by minimal residual disease, has allowed us to refine risk classification schemes and better tailor the intensity of therapy for each patient. More complete molecular analysis of leukaemia cells, pharmacodynamic and pharmacogenetic studies, and the development of targeted therapy should ultimately lead to further improvements in treatment. Pharmacogenetic studies should allow treatment refinements that will decrease the risk of complications while maintaining high cure rates. In addition, gene expression profiling may improve the genetic classification of leukaemia and identify clinically important subgroups. It may also lead to the identification of new targets for novel antileukaemic agents. PMID- 12633579 TI - Evidence-based oncology in cancer treatment reviews. PMID- 12633577 TI - Reirradiation of recurrent and second primary head and neck malignancies: a comprehensive review. AB - The management of locoregionally recurrent or second primary tumors in a previously irradiated head and neck volume presents a challenging clinical problem. Only a small subset of patients are candidates for potentially curative surgery. Chemotherapy alone provides only limited palliation with no long term survivors. Reirradiation, particularly with aggressive concomitant chemotherapy, results in prolonged median survival and long term survival for some patients. The toxicity of reirradiation, while greater than chemotherapy alone or primary irradiation, is lower than expected for the high cumulative radiation doses. The results of reirradiation in recurrent head and neck cancer and the prognostic factors predicting outcome in this patient population are reviewed. PMID- 12633580 TI - Evidence-based oncology section - aims and scope. PMID- 12633581 TI - Annual screening may not decrease breast cancer deaths among women aged 40 to 49 years. PMID- 12633582 TI - Uracil/tegafur plus oral leucovorin does not affect survival rates but may be better tolerated than standard fluorouracil and leucovorin for previously untreated metastatic colorectal cancer. PMID- 12633583 TI - Immediate thoracic radiotherapy may not improve symptom control over delayed radiotherapy in people with unresectable non-small cell lung cancer and minimal symptoms. PMID- 12633584 TI - Paclitaxel plus gemcitabine may be as active and well tolerated as paclitaxel plus carboplatin for advanced non-small-cell lung cancer. PMID- 12633585 TI - Immunochemotherapy with OK-432 may improve survival for people with curatively resected gastric cancer. PMID- 12633586 TI - Preserved ability to recognize keywords related to remote events in the absence of retrieval of relevant knowledge: a case of postencephalitic amnesia. AB - We describe a case of severe anterograde and retrograde amnesia resulting from herpes simplex encephalitis. Magnetic resonance imaging revealed pathological changes in the bilateral hippocampi, parahippocampal gyri, fusiform gyri, medial temporal poles, posterior part of the cingulate gyri, and insula. The patient showed severe amnesia for autobiographical episodic memory in relation to events that had occurred throughout her life, but temporally graded amnesia for autobiographical semantic memory, and severe amnesia without a temporal gradient for public events and famous people. However, using a multiple-choice method, she showed a high level of accuracy when choosing keywords related to public or personal events, although this did not prompt her recollection of the events. An important indication of these results is that, even with severe retrograde amnesia, memories of past events are not completely lost. We propose that an event may be stored in a fragmented form, consisting of many components, and that normal recall of an event may require recombination or reconstruction of these components. PMID- 12633587 TI - Developmental prosopagnosia: a study of three patients. AB - We studied perception in three patients with prosopagnosia of childhood onset. All had trouble with other 'within-category' judgments. All were deficient on face matching tests and severely impaired on tests of perception of the spatial relations of facial features and abstract designs, indicating a deficit in the encoding of coordinate relationships, similar to adult-onset prosopagnosia with lesions of the fusiform face area. Two had difficulty perceiving feature colour, which correlated with reduced luminance sensitivity. In contrast to adult-onset patients, saturation discrimination was spared in two and spatial resolution impaired in two. Curvature discrimination was relatively spared. Contrast sensitivity showed variable reductions at different spatial frequencies. We conclude that developmental prosopagnosia is similar to the adult-onset form in encoding deficits for the spatial arrangement of facial elements. Deficits in luminance perception and spatial resolution are more associated with defective encoding for basic object-level recognition, as shown on tests of object and spatial perception. PMID- 12633588 TI - Summation priming in aphasia: evidence for alterations in semantic integration and activation. AB - The present study used a lexical decision paradigm to study the summation of priming effects in normal and aphasic participants. The amount of priming produced by pairs of definitionally converging associative words was compared to the amount of priming produced by pairs of single associative words and non-words in two experiments in which the ISI between primes and targets varied from 200 ms (Experiment 1) to 600 ms (Experiment 2). Control subjects showed a pattern of additive summation priming at the short ISI and overadditive summation priming at the longer ISI. Broca's aphasics showed overadditive priming at the short ISI and no significant priming at the longer ISI; Wernicke's aphasics showed no significant priming at the short ISI and additive priming at the longer ISI. These results suggest that aphasics differ from normals in their ability to integrate the activation derived from multiple linguistic associations and may provide an account of some of the clinical phenomenology of these patients. PMID- 12633589 TI - Local-global processing in early-onset schizophrenia: evidence for an impairment in shifting the spatial scale of attention. AB - In this study we report the results of two experiments on visual attention conducted with patients with early-onset schizophrenia. These experiments investigated the effect of irrelevant spatial-scale information upon the processing of relevant spatial-scale information, and the ability to shift the spatial scale of attention, across consecutive trials, between different levels of the hierarchical stimulus. Twelve patients with early-onset schizophrenia and matched controls performed local-global tasks under: (1) directed attention conditions with a consistency manipulation and (2) divided-attention conditions. In the directed-attention paradigm, the early-onset patients exhibited the normal patterns of global advantage and interference, and were not unduly affected by the consistency manipulation. Under divided-attention conditions, however, the early-onset patients exhibited a local-processing deficit. The source of this local processing deficit lay in the prolonged reaction time to local targets, when these had been preceded by a global target, but not when preceded by a local target. These findings suggest an impaired ability to shift the spatial scale of attention from a global to a local spatial scale in early-onset schizophrenia. PMID- 12633590 TI - The vibrating nerve impulse in Newton, Willis and Gassendi: first steps in a mechanical theory of communication. AB - In later editions of his two major works, Isaac Newton proposed an electrical hypothesis of nervous transmission. According to this hypothesis, an electrical aether permeates the nerve and transmits vibrations along it. This implies that the nerve is a communication line, and potentially, an extension of the mind. The opposite view was held by Cartesian mechanists, who taught that the nerve is a power line, transmitting either pressure or tension, and that the mind is separate from the nervous system. The Newtonian model eventually supplanted the Cartesian model in the mid 18th century, and became a crucial part of the conceptual environment in which neuroscience originated. In this paper I examine the scientific origins of the Newtonian model of nervous transmission. I argue that Newton's model relies on prior work by Thomas Willis and Pierre Gassendi. Willis supplied the anatomical and physiological "hard data" upon which the model was built. But Gassendi, a generation before, laid out the conceptual foundations of the problem, including the principle of impulse-transmission, and the corrolary principle of the muscle as an autonomous generator of force. I conclude that Gassendi's work has been undeservedly neglected as a turning-point in the history of neuroscience. PMID- 12633591 TI - Representation of anatomical constraints in motor imagery: mental rotation of a body segment. AB - Classically, the mental rotation paradigm has shown that when subjects are asked to judge whether objects that differ in orientation are spatially congruent, reaction times increase with angular discrepancy, although some reports have shown that this is not always the case. Would similar results be obtained with realistic figures of body segments? In this work, the mental rotation of a hand attached to its forearm and arm in anatomically possible and impossible starting positions is compared with the mental rotation of a hammer. The main results show that reaction times increase monotonically with the angle of discrepancy for both stimuli and that the speed of rotation is higher for anatomically possible orientations in the case of the hand. Thus, mental rotation of body segments follows the same empirical rules as objects of another nature, and biomechanical constraints imposed to the motility of these segments can be considered as attributes of the mental representation. PMID- 12633592 TI - August Knoblauch and amusia: a nineteenth-century cognitive model of music. AB - Early models of human cognition can be traced to nineteenth-century investigations of brain and behavior. Influential neurologists such as Wernicke, Kussmaul, and Lichtheim constructed diagrammatic models to illustrate current theories of cognition. Language was the most commonly studied cognitive function during this time; however, investigators also studied other cognitive functions, such as music and visual processing. While a number of nineteenth-century neurologists made observations about music abilities in aphasic patients, August Knoblauch, a German physician and anatomist, was the first to propose a diagrammatic model of music (1888/1890). He described a detailed cognitive model of music processing, hypothesized the existence of nine disorders of music production and perception, and coined the term "amusia." Knoblauch's model is the earliest cognitive model of music and is largely unrecognized as an important part of the history of neurology, neuropsychology, and music cognition. PMID- 12633593 TI - Hemispheric asymmetries for global and local processing as a function of stimulus exposure duration. AB - The experiments assess the relationship between stimulus exposure duration and hemispheric asymmetries for global and local processing. Six durations (27, 40, 53, 67, 80, and 147 ms) were tested in a selective attention task in which hierarchical letterforms were presented unilaterally to the left or right visual field. The results indicated that left hemisphere-local asymmetries were demonstrated more often than right hemisphere-global asymmetries and that asymmetries were most commonly found in the middle range of durations tested. The pattern of results emphasizes the importance of further study into those conditions under which the predicted asymmetries can be reliably demonstrated in healthy individuals. PMID- 12633596 TI - A versatile assay for the accurate, time-resolved determination of cellular viability. AB - A convenient and versatile method for the accurate, time-resolved determination of cellular viability has been developed. The conventional viability indicator fluorescein diacetate (FDA), which is converted to the fluorescent compound fluorescein in living cells, was employed as a viability probe. Fluorescence emission from cells was measured using a spectrofluorimeter equipped with a magnetic stirrer. Using this assay cell suspensions exhibiting densities in the range 0.5 x 10(5) to 2.0 x 10(5) cells displayed a linear response when FDA concentrations less than 12 micro M were employed. To calibrate the method, viability standards were elaborated using different proportions of living and dead cells, and a correlation coefficient for the viability of tobacco BY-2 suspensions was calculated as 0.998. This viability assay was also found to be applicable to Chlamydomonas reinhardtii and Arabidopsis thaliana cultured cells. Using this cell viability assay, kinetic analyses of cell death could be performed. Using the proteinaceous elicitor from Phytophthora cryptogea, cryptogein, to induce cell death in tobacco cell suspensions, values for the maximum velocity of death induction rate (V(max)) and the LD50 (half-maximal velocity or k(1/2)) were calculated as 17.2 (% death/h) and 65 nM, respectively. PMID- 12633597 TI - Analysis of endocannabinoids by Ag+ coordination tandem mass spectrometry. AB - The neutral arachidonic acid derivatives N-arachidonylethanolamide (anandamide or AEA) and 2-arachidonylglycerol (2-AG) have been identified as endogenous ligands for the cannabinoid receptors. Quantitation of these endocannabinoids from various tissues has been shown to be essential in the elucidation of cannabinoid mediated processes in vivo. Here, we describe a novel method for the detection and quantitation of AEA and 2-AG from mammalian tissue. We exploit the ability of silver cation to bind to the polyunsaturated arachidonate backbone of both molecules to form the charged species [M+Ag](+). These complexes are amenable to liquid chromatography-electrospray ionization-tandem mass spectrometry analysis, resulting in the simple and specific quantitation of AEA and 2-AG. The limits of detection of 2-AG and AEA are 13 and 14fmol, respectively, on-column. This method provides an alternative to existing methods, which employ derivation and/or selected ion monitoring (when mass spectrometric detection is used), and may facilitate the understanding of the physiological roles of this new class of compounds. PMID- 12633594 TI - Effects of mental workload and caffeine on catecholamines and blood pressure compared to performance variations. AB - Caffeine is characterised as a central nervous system stimulant, also affecting metabolic and cardiovascular functions. A number of studies have demonstrated an effect of caffeine on the excretion of catecholamines and their metabolites. Urinary epinephrine and norepinephrine have been shown to increase after caffeine administration. Similar trends were observed in our study in adrenaline (ADR) and noradrenaline (NORADR) levels and additionally a dose dependent effect of caffeine. The effect of caffeine on cognitive performance, blood pressure, and catecholamines was tested under resting conditions and under mental workload. Each subject performed the test after oral administration of 1 cup and then 3 cups of coffee. Root mean square error (RMSE) for the tracking task was continuously monitored. Blood pressure was also recorded before and after each stage of the experiment. Catecholamines were collected and measured for three different conditions as: at rest, after mental stress alone, after one dose of caffeine under stress, and after triple dose of caffeine under stress. Comparison of the performance of each stage with the resting conditions revealed statistically significant differences between group of smokers/coffee drinkers compared with the other two groups of non-coffee drinkers/non-smokers and non smokers/coffee drinkers. There was no statistically significant difference between the last two groups. There was an increase of urine adrenaline with 1 cup of coffee and statistically significant increase of urine noradrenaline. Both catecholamines were significantly increased with triple dose of caffeine. Mental workload increased catecholamines. There was a dose dependent effect of caffeine on catecholamines. PMID- 12633598 TI - Development of an intact cell reporter gene beta-lactamase assay for G protein coupled receptors for high-throughput screening. AB - G protein-coupled receptors (GPCRs) are involved in a large variety of physiological disorders, and are thus important pharmaceutical drug targets. Here, we describe the development and characterization of a beta-lactamase reporter gene assay as a functional readout for the ligand-induced activation of the human bradykinin B1 receptor, expressed recombinantly in CHO cells. The beta lactamase reporter gene assay provides high sensitivity due to the absence of endogenous beta-lactamase activity in mammalian cells. The cell-permeable fluorogenic substrate allows single-cell cloning of cells expressing functional BK1 receptors. Pharmacological characterization reveals comparable sensitivity and potency of known BK1 receptor agonists and antagonists between the beta lactamase assay, competition-binding assay, and other direct measurements of second messengers. The beta-lactamase assay has been optimized for cell density, time of agonist stimulation, and DMSO sensitivity. This CHO-hBK1-beta-lactamase assay is well suited to automation and miniaturization required for high throughput screening. PMID- 12633599 TI - Classification of DNA-binding mode of antitumor and antiviral agents by the electrochemiluminescence of ruthenium complex. AB - The DNA-binding mode of antitumor and antiviral agents has been evaluated by electrochemiluminescence (ECL) of tris(1,10-phenanthroline)-ruthenium complex (Ru(phen)(3)(2+)) in the presence of oxalate ion in pH 7.3 Tris buffer solution. An emission of Ru(phen)(3)(2+) was observed repeatedly with a voltage above 1000mV subjected to a potential sweep from 0 to 1250mV. The addition of lambdaDNA into the solution containing 1 micro M of Ru(phen)(3)(2+) caused the decrease in the ECL intensity, which became half at a DNA concentration of 20 micro M. This is due to the binding of Delta-type of Ru(phen)(3)(2+) with DNA in the major groove of DNA. When the various concentrations of the drug were added to the solution containing 1& micro M Ru(phen)(3)(2+), the ECL intensity was not affected by the concentration of the drug in the absence of DNA. In the presence of DNA (10 micro M), however, two ECL emission patterns were observed when the concentration of the drug was varied. The pattern that the ECL intensity increased with increasing the drug concentration was observed for cisplatin, daunomycin, and DC92-B. This may have resulted from the DNA binding of the drug with a major groove site, where Ru(phen)(3)(2+) should bind. Ru(phen)(3)(2+) nonbinding to DNA might exist in the bulk solution and exhibits ECL emission. The drug exhibiting the drug-concentration-dependent ECL is classified as a drug with a major groove binding character. The addition of drugs, such as mitomycin C and duocarmycin SA, did not cause a change in the ECL intensity even in the presence of DNA. This result indicates that these drugs bind to DNA with minor groove binding. Since similar trends were observed for actinomycin D, distamycin A, doxorubicin, and chromomycin A3; these drugs are also considered as minor groove binding agents. All these results demonstrate that the DNA-binding mode of the drug can be evaluated easily by utilizing the ECL of Ru(phen)(3)(2+), which is used as the sensing probe. PMID- 12633600 TI - Development and characterization of simulant pancreatic islets. AB - Insulin is stored in pancreatic islets as a zinc-insulin complex, and stimulating the islets results in the release of insulin and zinc. Simulant pancreatic islet beads have been developed using agarose beads (50-250 micro m diameter) derivatized with iminodiacetic acid that have been loaded with zinc. A qualitative comparison of the simulant beads with pancreatic islets has been made by staining with dithizone and a zinc-binding fluorescent dye, TSQ. The binding capacity of simulant beads was determined to be 34 micro mol Zn(2+)/g of dried beads using anodic stripping voltammetry. Hydrochloric acid was used to release zinc from beads to mimic the secretion of insulin from pancreatic islets and a release profile was established. The simulant beads can be used to optimize the islet isolation process and reduce the use of real islets in method development. PMID- 12633601 TI - Downsizing improves sensitivity 100-fold for hydrogen exchange-mass spectrometry. AB - This paper describes a method that substantially improves the sensitivity of high performance liquid chromatography hydrogen exchange-mass spectrometry (HPLC HX MS). The success of this method relies on using a capillary HPLC column (0.1mm IDx5cmL) to increase the sensitivity of electrospray ionization, while keeping analysis times short to minimize hydrogen/deuterium (H/D) exchange. A small, immobilized pepsin column and a capillary C18 trap were included in the capillary HPLC MS system to provide rapid digestion, peptide concentration, and desalting while maintaining slow H/D exchange conditions. To minimize the analysis time, dead volumes and capacities of all components were optimized. Fully deuterated cytochrome c and its fully deuterated peptic peptides were used to evaluate deuterium recovery at amide linkages. The deuterium recovery measured at low flow rates using this system spanned a range of 66-77% (average of 71%), which was similar to the range measured for a much larger system (67-80%, average 75%). Signal levels of most peptides for the downsized system increased by about 100 fold compared with the signal for the larger system. These results greatly strengthen the HPLC HX MS technique for studies where the quantity of protein is small. PMID- 12633602 TI - Multiplex single-nucleotide polymorphism genotyping by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - A robust high-throughput single-nucleotide polymorphism (SNP) genotyping method is reported, which applies allele-specific extension to achieve allelic discrimination and uses matrix-assisted laser desorption/ionization time-of flight mass spectrometry to measure the natural molecular weight difference of oligonucleotides for determination of the base in a single-nucleotide polymorphic location. Tenfold PCR is performed successfully by carefully designing the primers and adjusting the conditions of PCR. In addition, two ways used for PCR product purification are compared and the matrix used in mass spectrometry for high-throughput oligonucleotide analysis is evaluated. The result here shows that the method is very effective and suitable for high-throughput genotyping of SNPs. PMID- 12633603 TI - Determination of sugar phosphates and nucleotides related to photosynthetic metabolism by high-performance anion-exchange liquid chromatography with fluorometric and ultraviolet detection. AB - A high-performance anion-exchange liquid chromatography system was constructed to identify sugar phosphates and nucleotides involved in photosynthetic metabolism. First sugar phosphates and nucleotides were separated by a gradient elution with boric acid and sodium phosphate, then they were detected by a fluorescence detector (as fluorescent derivatives with arginine) and UV detector, respectively. Eight authentic sugar phosphates and 11 authentic nucleotides could be analyzed using the system. The applicability of the system to the determination of the corresponding sugar phosphates and nucleotides in extracts from only five soybean leaf discs (8.95 cm2) was shown. PMID- 12633604 TI - Cationic electrophoresis and electrotransfer of membrane glycoproteins. AB - Sodium dodecyl sulfate electrophoresis of glycoproteins results in broad, fuzzy bands. The use of the positively charged detergent cetyltrimethylammonium bromide (CTAB) avoids this problem. Protocols for CTAB electrophoresis, electroblotting, and protein detection are presented. These methods were found useful in studies on the multiple drug resistance ATPase Mdr1. PMID- 12633605 TI - Intercalation activating fluorescence DNA probe and its application to homogeneous quantification of a target sequence by isothermal sequence amplification in a closed vessel. AB - We developed a completely homogeneous and isothermal method of detecting RNA sequences and demonstrated ultrarapid and direct quantification of pathogenic gene expression with high sensitivity. The assay is based on performing isothermal RNA sequence amplification in the presence of our novel DNA probe, an intercalation activating fluorescence DNA probe, and measuring the fluorescence intensity of the reaction mixture. When detecting mecA gene expression of methicillin-resistant Staphylococcus aureus, we quantified starting copies ranging from 10 to 10(7) copies within 10min. The primer sequences were designed to bind to secondary structure-free sites of the target RNA, which enabled a totally isothermal protocol to quantify mRNA specifically in a sample of existing genomic DNA. When we applied this to quantifying the expression of marker genes of Vibrio parahaemolyticus and Mycobacterium bovis BCG strain, the results correlated well with the viability of each bacterium. We also demonstrated monitoring Pab gene expression of M. bovis BCG during cultivation with antibiotics. The present method can potentially realize rapid antimicrobial susceptibility testing of slowly growing organisms, such as tuberculosis. PMID- 12633606 TI - Sensitive time-resolved fluoroimmunoassay for simultaneous detection of serum thyroid-stimulating hormone and total thyroxin with Eu and Sm as labels. AB - Based on a novel cocoating strategy and dissociation enhancement lanthanide fluorescence immunoassay technique, a sensitive time-resolved fluoroimmunoassay (TRFIA) has been developed for simultaneous quantification of human serum thyroid stimulating hormone (TSH) and thyroxin (T4) in a one-and-the-same assay procedure. The new cocoating strategy for preparing highly active surface anti TSH and anti-T4 monoclonal antibodies (McAbs) was performed by a three-step protocol. Namely, anti-TSH McAb at high concentration (10 micro g/ml) and extensively biotinylated bovine serum albumin (BSA) at low concentration (0.5 micro g/ml) were coated on microwells by passive adsorption, then streptavidin was captured by the surface BSA-biotin, and finally biotinylated anti-T4 McAb was immobilized by the remnant binding sites of the bound streptavidin. In the present TSH/T4 TRFIA, both sandwich- and competitive-type configurations were involved, and Eu(3+) and Sm(3+) were used as labels for TSH and T4 detection, respectively. The method showed rapid kinetics; the equilibrium was reached within 30min at 37 degrees C due to the use of high concentrations of reaction reagents, rapid agitation, and small reaction volume. The lower limits of detection of the method were 0.028 mIU/L for TSH and 4.1 nmol/L for T4 with 20 micro L of sample volume. The assay ranges for TSH and T4 were 0.21-80.00 mIU/L and 20-300 nmol/L, respectively. The correlation between the TSH/T4 values obtained by the present TSH/T4 TRFIA and those obtained by commercial chemiluminescence immunoassay was satisfactory. PMID- 12633607 TI - Separation of phospholipids in microfluidic chip device: application to high throughput screening assays for lipid-modifying enzymes. AB - Phospholipid molecules such as ceramide and phosphoinositides play crucial roles in signal transduction pathways. Lipid-modifying enzymes including sphingomyelinase and phosphoinositide kinases regulate the generation and degradation of these lipid-signaling molecules and are important therapeutic targets in drug discovery. We now report a sensitive and convenient method to separate these lipids using microfluidic chip-based technology. The method takes advantage of the high-separation power of the microchips that separate lipids based on micellar electrokinetic capillary chromatography (MEKC) and the high sensitivity of fluorescence detection. We further exploited the method to develop a homogenous assay to monitor activities of lipid-modifying enzymes. The assay format consists of two steps: an on-plate enzymatic reaction using fluorescently labeled substrates followed by an on-chip MEKC separation of the reaction products from the substrates. The utility of the assay format for high-throughput screening (HTS) is demonstrated using phospholipase A(2) on the Caliper 250 HTS system: throughput of 80min per 384-well plate can be achieved with unattended running time of 5.4h. This enabling technology for assaying lipid-modifying enzymes is ideal for HTS because it avoids the use of radioactive substrates and complicated separation/washing steps and detects both substrate and product simultaneously. PMID- 12633608 TI - Flow cytometric and immunoblot assays for cell surface ADP-ribosylation using a monoclonal antibody specific for ethenoadenosine. AB - NAD-dependent ADP-ribosylation is one of the posttranslational protein modifications. On mammalian cells, glycosylphosphatidylinositol-anchored cell surface ADP-ribosyltransferases (ARTs) ADP-ribosylate other cell surface proteins and thereby affect important cellular functions. Here we describe convenient flow cytometric and immunoblot assays for monitoring ADP-ribosylation of cell surface proteins on living cells by exploiting the capacity of ARTs to utilize etheno-NAD as substrate. Etheno-ADP-ribosylation of cell surface proteins can be detected by flow cytometry with 1G4, a monoclonal antibody specific for ethenoadenosine. Labeling of cells with 1G4 is dependent on the expression of cell surface ARTs and occurs only after incubation of ART-expressing cells with etheno-NAD and not with etheno-ADP-ribose. Dose-response analyses show efficient 1G4 staining of ART expressing cells at micromolar etheno-NAD concentrations. Half-maximal staining is obtained with 1-2 micro M etheno-NAD, saturation is reached at 5-20 micro M etheno-NAD. Immunoblot analyses confirm that ART-expressing cells incorporate ethenoadenosine covalently (i.e., SDS resistant) into several cell surface proteins. The flow-cytometric 1G4 staining assay can be used to identify subpopulations of cells expressing cell surface ART activity and to select ART(hi) cell variants. The immunoblot 1G4 staining assay can also be used to identify etheno-ADP-ribosylated target proteins. These new assays hold promise for many interesting applications in biochemistry and cell biology. PMID- 12633609 TI - A fluorescence-based, high-performance liquid chromatographic assay to determine acid sphingomyelinase activity and diagnose types A and B Niemann-Pick disease. AB - Acid sphingomyelinase (ASM; sphingomyelin phosphodiesterase, EC 3.1.4.12) is the lysosomal enzyme that hydrolyzes sphingomyelin (SPM) to phosphorylcholine and ceramide. An inherited deficiency of ASM activity results in Types A and B Niemann-Pick disease (NPD). In this study we report a new assay method to detect ASM activity and diagnose NPD using the fluorescent substrate BODIPY C12-SPM and reverse-phase high-performance liquid chromatography (HPLC). The reaction product, BODIPY C12-ceramide (B12Cer), could be clearly and efficiently separated from the substrate within 4 min using a reverse-phase column (Aquasil C18, Keystone Scientific). Femtomole quantities of B12Cer could be detected in as little as 1.0 micro l of human plasma, providing a sensitive measure of ASM activity. The mean ASM activity in human plasma from NPD patients (36 pmol/ml/h) was only 2.7% of that in normal plasma (1334 pmol/ml/h), confirming the specificity and diagnostic value of this new assay method. Importantly, the mean ASM activity in human plasma from NPD carriers (258.3 pmol/ml/h) also was significantly reduced (19.5% of normal). The ranges of ASM plasma activities in NPD patients (N=19), NPD carriers (N=11), and normal subjects (N=15) were 2.5 97.3, 108-551, and 1030-2124 pmol/ml/h, respectively. Based on these results, we suggest that this fluorescence-based HPLC assay method is a reliable, rapid, and highly sensitive technique to determine ASM activity and that plasma is a very reliable and simple source for the accurate diagnosis of NPD patients and carriers based on ASM activity. PMID- 12633610 TI - Immunocapture and microplate-based activity measurement of mammalian pyruvate dehydrogenase complex. AB - Altered pyruvate dehydrogenase (PDH) functioning occurs in primary PDH deficiencies and in diabetes, starvation, sepsis, and possibly Alzheimer's disease. Currently, the activity of the enzyme complex is difficult to measure in a rapid high-throughput format. Here we describe the use of a monoclonal antibody raised against the E2 subunit to immunocapture the intact PDH complex still active when bound to 96-well plates. Enzyme turnover was measured by following NADH production spectrophotometrically or by a fluorescence assay on mitochondrial protein preparations in the range of 0.4 to 5.0 micro g per well. Activity is sensitive to known PDH inhibitors and remains regulated by phosphorylation and dephosphorylation after immunopurification because of the presence of bound PDH kinase(s) and phosphatase(s). It is shown that the immunocapture assay can be used to detect PDH deficiency in cell extracts of cultured fibroblasts from patients, making it useful in patient screens, as well as in the high-throughput format for discovery of new modulators of PDH functioning. PMID- 12633611 TI - Separation of saccharides derivatized with 2-aminobenzoic acid by capillary electrophoresis and their structural consideration by nuclear magnetic resonance. AB - Saccharides including mono- and disaccharides were quantitatively derivatized with 2-aminobenzoic acid (2-AA). These derivatives were then separated by capillary zone electrophoresis with UV detection using 50mM sodium phosphate buffer as the running electrolyte solution. In particular, the saccharide derivatives with the same molecular weight as 2-AA aldohexoses (mannose and glucose) and 2-AA aldopentoses (ribose and xylose) were well separated. The underlying reasons for separation were explored by studying their structural data using 1H and 13C NMR. It was found that the configurational difference between their hydroxyl group at C2 or C3 could cause the difference in Stokes' radii between their molecules and thus lead to different electrophoretic mobilities. The correlation between the electrophoretic behavior of these carbohydrate derivatives and their structures was studied utilizing the calculated molecular models of the 2-AA-labeled mannose, glucose, ribose, and xylose. PMID- 12633612 TI - A spectrophotometric assay for measuring and detecting an epoxide hydrolase activity. AB - In this paper we report the development of a novel and simple spectrophotometric assay which allows one to achieve the continuous, rapid, sensitive, and accurate determination of an epoxide hydrolase activity. This assay is based on the elaboration of a coupled enzymatic/chemical methodology which allows quantification of the enzymatic activity within 3min, and offers good sensitivity of about 10 micro Mmin(-1). Applicability of this test to some other aromatic epoxides has been shown and some limitations have also been explored. This assay should be particularly useful for different applications, for example (a) activity localization during purification of such enzymes, (b) very rapid determination of kinetic constants, and (c) high-throughput screening experiments. PMID- 12633613 TI - Modified protocol for hepatitis B virus DNA isolation and detection. PMID- 12633614 TI - Rapid detection of CSRP2 mRNA in mouse, rat, and human using LightCycler-based quantitative real-time polymerase chain reaction. PMID- 12633615 TI - Electrospray ionization tandem mass spectrometry scan modes for plant chloroplast lipids. PMID- 12633616 TI - Application of electrochemical quartz crystal microbalance technique to direct monitoring of cytochrome c3 function as the electron pool during intermolecular electron transfer. PMID- 12633617 TI - Method enabling firefly luciferase-based bioluminometric assays at elevated temperatures. PMID- 12633618 TI - Receptor binding assay for nitric oxide- and heme-independent activators of soluble guanylate cyclase. PMID- 12633619 TI - 4-(Aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole is not specific for labeling of sulfhydryl groups in proteins as it may also react with phenolic hydroxyl groups and amino groups. PMID- 12633620 TI - A nurturing environment. PMID- 12633621 TI - Activated charcoal: the untold story. AB - Introduction. To identify the prevalence and appropriateness of prescribing activated charcoal in the management of acute poisoning and to document patient compliance with treatment.Methods. A prospective study was conducted, between October 1998 and September 1999, on patients attending our accident and emergency department, with a history of overdose. Overdoses were classified as potentially toxic or non-toxic according to the history and/or information received from the National Poisons Information Service.Results. Two hundred and seventy five patients presented following overdose; 17% within one hour, 102 were prescribed charcoal (37.1%) but of these, 40 (39%) refused it, and of the 62 patients (61%) who accepted charcoal only 15 (24.2%) took all that was prescribed. Patients were 5.4 times more likely to take charcoal if they had taken a potentially toxic overdose. Of those who presented within one hour and were judged to have taken a potentially toxic overdose, only three patients took the full-prescribed amount.Conclusion. We report a substantially greater proportion of patients (39%) refusing charcoal than previously reported (9.9%). The widespread availability of TOXBASE Copyright should help redress this discrepancy. PMID- 12633622 TI - The politics of beds. PMID- 12633623 TI - Adolescents accessing emergency contraception in the A&E department - a feminist analysis of the nursing experience. AB - This paper outlines the findings and discussion of a qualitative research study that focused on the experiences of seven qualified nurses working in three Accident and Emergency (A&E) departments in the North West of England. It was exploratory in nature, aiming to describe and explain the nurses' encounters with adolescents accessing emergency contraception (EC) in A&E. The study was carried out using a feminist methodology and a grounded theory method. The findings indicated that accident and emergency is a contradictory location for access to EC for adolescents, where the 'promise' of easy, confidential access contrasts sharply with the nurse's description of reality. The nurse's role is similarly contrasting, where their ideal is counter balanced by organisational limits, and is further shaped by both personal and professional guiding philosophies. The nurse's perceptions of the adolescents revealed the contradiction of both sympathetic and judgmental attitudes towards them, including an 'interpretation' of the reasons the adolescents gave for their attendance. These encounters led to a series of health, legal, and moral dilemmas for the nurses, and a strategy of referral of the adolescents to other agencies was used by them whenever possible. PMID- 12633624 TI - Where have all the bubbles gone? An ode to Hydrogen peroxide, the champagne of all wound cleaners. AB - Hydrogen peroxide solution can be very effective for cleaning grit filled wounds and grazes. Its use in Accident and Emergency (A&E) departments has gone out of vogue due to concerns about air emboli formation. This article discusses the truth behind these concerns and whether it is in fact safe to use hydrogen peroxide solution for wound cleaning in specific situations. PMID- 12633625 TI - Establishing a bereavement program: caring for bereaved families and staff in the emergency department. AB - The successful planning and implementation of a Bereavement Program in the Emergency Departments of two of Melbourne's major metropolitan teaching hospitals, Monash Medical Centre (MMC) and Dandenong Hospital (DH) was in response to staff dissatisfaction regarding their capacity to care for bereaved families and involved staff. Key influencing factors were identified and literature evidence was sought. A multidisciplinary team was brought together to form a Planning Group and guidelines were developed. The aim was to improve the care of all those involved with a death in the Emergency Department, both family and staff. Fundamental to facilitating these improvements was the need for staff education in the needs of families, self and staff and a grief education program was arranged, and provided through multiple sources, utilising the expertise of the Centre for Grief Education. The extremely positive feedback, which has been received from families and ED staff is testimony to the care delivered and the difference the Program is making at Southern Health, to those experiencing a death in the Emergency Department. PMID- 12633626 TI - Female catheterisation: what nurses need to know! AB - Urinary catheter insertion is a common procedure used in the management of hospitalised and community based patients. Nurses need to be aware of the associated complications of catheterisation in order to practice safely and minimise the risks to the patient. This article is a practical guide for nurses who will be involved in female catheterisation, from decision to catheterise, choosing the type and size of catheter, and tips on catheter care and how to avoid the most common problems associated with catheterisation. PMID- 12633627 TI - Developing expertise - the contribution of paediatric accident and emergency nurses to the care of children, and the implications for their continuing professional development. AB - AIM: The purpose of this paper is to explore the contribution of paediatric accident and emergency nurses to the care of children. METHOD: Using a case study approach, the paper focuses on the experiences of three nurses working in this setting, obtained through working alongside the nurses and discussing and observing their experiences. FINDINGS: It became apparent that the nurses were regarded as "experts" in the care of children in this setting. The nurses themselves however, did not share this perception. Their experiences are analysed with reference to literature on expertise, focussing specifically on published accounts of children's nurses' experiences, and research which has explored the concept of expertise within the context of accident and emergency work. The implications for continuing professional development needs are discussed within the context of recent government reports, which make recommendations in respect of access and delivery of on-going professional education. To this end it is advocated that a multi-professional approach is adopted, with children's nurses sharing learning and thus access to, courses customarily provided to adults nurses working in this setting. PMID- 12633628 TI - A case report and review of the locked metacarpophalangeal joint. AB - A 59-year-old female with metacarpal joint locking is presented. The successful closed manipulation is described with a review of the literature. PMID- 12633629 TI - Snake bite in Australia: first aid and envenomation management. AB - Australia is inhabited by a large variety of snakes, including some of the most poisonous in the world. Sightings are regular, and the incidence of snake bite is estimated to be several thousand a year. A bite does not necessarily result in envenomation occurring, however there are at least 300 snakebites a year requiring treatment of envenomation, with between 1 and 4 fatalities every year. The incidence of fatalities from snake bite has increased over recent years. The explanation for this is unclear, but possible reasons include the urban sprawl, and a delay in application of appropriate first aid and definitive treatment for envenomation. Emergency nurses in particular should be aware of the first aid techniques appropriate for Australian snake bite, as well as the recognition and management of envenomation. This article will outline the steps required and rationale for applying first aid techniques considered to be effective in retarding spread and circulation of snake venom. It will also discuss the manifestations that indicate systemic envenomation, and management considered to be responsible for reducing the incidence of death from snake envenomation in Australia. PMID- 12633631 TI - The introduction of the Manchester triage scale to an emergency department in the Republic of Ireland. AB - Triage is an integral part of the modern emergency department. The use of a recognised triage system has many advantages for the emergency department including reference to a recognised decision-making structure and support in the form of a professionally accepted and validated system. As part of a programme of internal change the Manchester triage system (MTS) was introduced to an emergency department in the Republic of Ireland. This article outlines the introduction of this method of triage and cites the domestic and international drivers of the change. PMID- 12633630 TI - Nursing issues in care for the elderly in the emergency department: an overview of the literature. AB - Emergency care is an important link in the chain of care for geriatric patients. Due to the specific physiological, psychological, and social needs of elderly people, the type and quantity of emergency care are different for this group of patients than for other age groups. Consequently, they have a significant impact on the health care system. Doctors and nurses working in an emergency department must be aware of this specific situation and adjust the care that they provide accordingly. Appropriate communication between health care providers and patients, a sound assessment of functional status and cognitive ability, and specific attention to verbal and written discharge guidelines are indispensable. PMID- 12633634 TI - International consensus on nomenclature and classification of atrial fibrillation; a collaborative project of the Working Group on Arrhythmias and the Working Group on Cardiac Pacing of the European Society of Cardiology and the North American Society of Pacing and Electrophysiology. PMID- 12633635 TI - Pacemaker prevention therapies for the control of drug-refractory paroxysmal atrial fibrillation. AB - AIMS: Atrial septal pacing has been shown to prevent paroxysmal atrial fibrillation (PAF) refractory to drugs in patients without inappropriate bradycardia. This study assesses the effects of atrial septal pacing using new pacing algorithms designed to prevent the initiation or maintenance of PAF. METHODS AND RESULTS: Eleven Medtronic AT500 and 6 Guidant Pulsar Max pacemakers were implanted. The incremental benefit of prevention pacing therapies was compared with DDDR pacing by analysis of pacemaker-stored electrograms, ambulatory electrocardiography, symptoms and quality of life questionnaires. RESULTS: Atrial septal pacing reduced AF burden by >50% in 13/17 patients (76.5%). Activation of a combined pacing algorithm (atrial pacing preference; atrial rate stabilization; and post mode-switch overdrive pacing) in patients with AT500 pacemakers produced a marginal reduction in AF burden (mean %AF 0.61 ON, 0.73 OFF, P=0.53 ns). Conversely in the Pulsar Max group when atrial pacing preference was activated, AF burden was slightly increased (mean %AF 5.84 ON, 3.73 OFF,P =0.13). Symptoms improved with atrial septal pacing but did not change when prevention algorithms were activated. CONCLUSION: Atrial septal pacing resulted in a marked improvement in AF burden and symptoms. Activation of specific prevention pacing algorithms provided more continuous atrial pacing but had limited and heterogeneous effects on AF burden. PMID- 12633636 TI - Supraventricular arrhythmia before and after surgical closure of atrial septal defects: spectrum, prognosis and management. AB - Supraventricular arrhythmias are often observed in patients before and after atrial septal defect repair. Although several papers report different incidences of sustained supraventricular arrhythmias, postoperative 'incisional' macroreentrant tachycardias have not been systematically investigated. METHODS: We reviewed 136 consecutive patients (79 female, 57 male, mean age 36.8+/-17.8 years) who underwent atrial septal defect repair at our institutions between January 1990 and January 1999. Coexisting valve disease requiring surgical intervention was noted in 13 patients (9.5%). The mean follow-up period was 78.8+/-30.1 months. RESULTS: Sustained supraventricular arrhythmias occurred in 12 patients (8.8%) before surgery (atrial fibrillation in 11 patients). Using multivariate analysis the occurrence of arrhythmia significantly correlated with the presence of coexisting heart disease (P< 0.001) and age at surgery (P=0.011) After surgery sustained supraventricular arrhythmias were recorded in 16 patients (11.7%). Eleven of them had atrial fibrillation, permanent in 8 cases, 4 'incisional' macroreentrant atrial tachycardia and 1 atrioventricular re-entry tachycardia. There was a significant correlation between pre and postoperative arrhythmia (P< 0.001). Two of the 4 patients with macroreentrant atrial tachycardia underwent successful radiofrequency catheter ablation, whereas the arrhythmia was controlled medically in the remaining 2 patients. CONCLUSIONS: Atrial fibrillation remains the most frequent form of arrhythmia before and after surgical closure of atrial septal defects in adulthood, and relates to age at the time of repair and coexisting heart disease. Incisional macroreentrant atrial tachycardia is an identifiable, albeit less common, form of tachycardia, which can be treated by transcatheter ablation. PMID- 12633637 TI - Home monitoring in ICD therapy: future perspectives. AB - The expanding indications for ICD therapy and the complexity of current devices will have impact on follow-up policy. The application of ICD therapy requires elaborate attention to technical aspects, arrhythmias, and the clinical course of the underlying disease. Currently, the quality of medical supervision is dependent on scheduled regular follow-up visits. A disadvantage of long intervals can be a delay in the physician's or patient's awareness of changes in the clinical status. Some patients will need more intensive follow-up while others will have the device as a stand-by and only need technical follow-up. A possibility to address this situation, is the transmission of data, already stored in the implanted device. This will guarantee continuous patient surveillance and could possibly help to avoid unnecessary follow-up visits. PMID- 12633639 TI - Glossopharyngeal neuralgia with cardiac syncope treated by glossopharyngeal rhizotomy and microvascular decompression. AB - A glossopharyngeal neuralgia case with cardiac asystole is presented. The sinus mode dysfunction and subsequent syncope with pain appears to be the most important life-threatening symptom in the late period of the disease. Because of cardiac symptoms induced by intense vagal stimulation, this case was considered to be vagoglossopharyngeal neuralgia. Several medical and surgical alternatives have been proposed for its treatment. In the presented case the cause of disease was compression of lower cranial nerves in the right cerebello-pontine angle (CPA) by the vertebro-basilar arterial system. Carbamazepine and pacing were determined ineffective, so the patient was treated surgically by cutting the glossopharyngeal nerve and by decompression of the vagal nerve at the CPA. The sinus arrest and paroxysmal pain attacks disappeared and the patient's life returned to normal. PMID- 12633638 TI - Sensing issues related to the clinical use of implantable loop recorders. AB - AIMS: The newer insertable loop recorder device (Reveal Plus 9526, Medtronic Inc, Minneapolis MN, U.S.A.) is equipped with auto-activation capabilities. In this study we investigated whether the new device encounters sensitivity problems in the form of oversensing or undersensing. We also tested whether body position changes influence the sensed electrogram's amplitude, therefore affecting the device's autosensing ability. METHODS AND RESULTS: We enrolled 32 patients aged 58.1+/-11.7 years with a loop recorder implanted either in the left parasternal or in the heart's apex area, to investigate undiagnosed syncopal episodes after initial assessment. During the follow-up period (5.7+/-2.7 months), that began 6 months after the device implantation, we analysed 284 auto-recorded episodes (103 undersensed and 181 correctly sensed). No oversensing episodes were recorded. THE RECORDED QRS amplitude in five different body positions: supine, left and right lateral, sitting and standing was measured. No statistically significant changes were observed in the different postures. Even when patients were divided into subgroups according to implant site or the occurrence of undersensing episodes, no significant inter- or intra-group differences of the sensed electrogram were observed. CONCLUSION: Undoubtedly this device is the current gold standard for the diagnosis of unexplained syncope. In the population we studied though, the device faced undersensing problems. Consequently, the device should be equipped with more advanced software for QRS morphology recognition, to improve the diagnostic accuracy of the recorder. PMID- 12633640 TI - The DDDR closed loop stimulation for the prevention of vasovagal syncope: results from the INVASY prospective feasibility registry. AB - BACKGROUND: The contraction dynamics of the ventricular myocardium are affected before and during vasovagal fainting suggesting that the Closed Loop Stimulation (CLS) pacemaker could be useful for the treatment of these patients. CLS is a new concept of heart rate modulation in cardiac pacing. The pacemaker INOS(2) CLS (Biotronik, Germany) derives its information for heart rate optimization from myocardial contraction dynamics, by measuring right ventricular intracardiac impedance. The pacemaker becomes an integral part of the circulatory regulation and, therefore, reacts appropriately to different cardiovascular demands. METHODS: In a prospective registry, 34 patients with a history of recurrent vasovagal syncopal events were implanted with INOS(2) DDDR CLS pacemakers. The aim of the study was to evaluate both long term clinical outcome, including the first recurrence of syncope, with DDDR-CLS pacing and acute precipitation of vasovagal fainting with DDDR-CLS mode compared with DDD during head up tilt testing. RESULTS: During a follow up period of 12-50 months, 30 patients experienced no further syncopal events in daily life; 1 patient had no syncope but night palpitations, which were eliminated by pacemaker reprogramming; 2 patients had presyncopal episodes but not syncopes; 3 syncopal recurrences occurred in one patient in chronic atrial fibrillation, possibly not an ideal candidate for implantation. CONCLUSIONS: Further studies for detailed understanding of the preventive mechanism of DDDR-CLS pacing in vasovagal syncope are warranted. A randomized multicentre prospective new study (INotropy controlled pacing in VAsovagal SYncope: INVASY) is now in progress to confirm the beneficial effect of DDDR-CLS pacing in a larger group of patients with recurrent vasovagal syncope. PMID- 12633641 TI - Characterization of the non-linear rate-dependency of QT interval in humans. AB - AIMS: Repolarization has rate-dependent and rate-independent components. A function considering such components separately was validated in canine Purkinje fibres and applied to the QT/RR relation in humans. METHODS AND RESULTS: Action potential duration (APD) was measured in Purkinje fibres during steady-state pacing at different cycle lengths (CL) and after prolonged quiescence (APD(0)). The APD/CL relationship was expressed by this function: APD=APD(max)(*)CL(S)/(CL(50)(S)+CL(S)), where APD(max) (APD extrapolated at infinite CL) is a rate-independent measure of repolarization, CL(50) (CL at which 50% of APD(max) is achieved) and S evaluates the rate dependency of APD. The same function was used to fit the QT/RR relation in 46 normal subjects (20 males, 26 females) and in 7 amiodarone-treated subjects undergoing a bicycle stress test. RR and QT (V(5)) were measured at the end of each load step; QT(c) (Bazett's formula) was obtained at rest. The APD/CL and QT/RR relations were equally well expressed by the function with high correlation coefficients (R>or=0.90). In Purkinje fibres, APD(max) was 461+/-37 ms, CL(50) was 394+/-54 ms and S was 0.98+/-0.11. APD(max) and APD(0) correlated (R=0.96) and were similar. The corresponding values in humans were: QT(max) 432+/-63 ms, RR(50) 345+/-60 ms and S 2.6+/-0.8. While QT(c) and QT(max) were longer in females, RR(50) and S were similar between genders. Amiodarone increased QT(c), QT(max) and RR(50) and decreased S. In QT(max) and QT(c) distributions generated by pooling data from treated and untreated subjects, 86% of treated subjects were correctly identified by QT(max) and 28% by QT(c). CONCLUSIONS: Canine and human repolarization showed a saturating dependency on cycle length, described by the proposed function. Gender and amiodarone independently affected QT(max), RR(50) and S: therefore they might reflect specific ionic mechanisms. Finally, QT(max) identified drug induced repolarization abnormalities in individual subjects better than QT(c). PMID- 12633642 TI - Slow pathway catheter ablation of atrioventricular nodal re-entrant tachycardia guided by electroanatomical mapping: a randomized comparison to the conventional approach. AB - BACKGROUND: Electroanatomical mapping may be expected to improve safety, efficiency and efficacy of selective slow pathway ablation for atrioventricular nodal re-entrant tachycardia (AVNRT). The goal of this prospective randomized study was to compare the efficiency of conventional fluoroscopic and electroanatomical mapping in guiding catheter ablation of AVNRT. METHODS AND RESULTS: Following induction of typical AVNRT, 20 consecutive patients were randomized to either conventional fluoroscopic or electroanatomical (CARTO) mapping to guide slow pathway ablation using a 4mm electrode. Endpoints for ablation were non-inducibility and no more than a single AV nodal echo on aggressive retesting. Acute procedural success was 100% in both groups, with no complications. Although there were no differences in time taken for pre- and post ablation electrophysiological evaluations, in the electroanatomical group the ablation portion of the procedure showed a substantial reduction in duration (12.6+/-6.8 vs 35.9+/-18.3 min; P< 0.001) and fluoroscopic exposure (0.7+/-0.5 vs 9.6+/-5.0 min; P< 0.001) compared with the fluoroscopic group, reflected in reduced total procedure time (83.6+/-23.6 vs 114+/-19.3 min; P=0.008) and total fluoroscopic exposure (4.2+/-1.4 vs 15.9+/-6.4 min; P< 0.001). Electroanatomical mapping was associated with a lower number (2.7+/-1.6 vs 5+/-2.8; P=0.018), duration (165.3+/-181.6 vs 341+/-177.7s; P=0.013), and total energy delivery (24.3+/-3.1 vs 28.7+/-4.5 watts; P=0.042) of RF applications. There were no acute or long-term (8.9+/-2.2 month) complications or arrhythmia recurrence in either group. CONCLUSIONS: While both conventional and non-fluoroscopic electroanatomical mapping are associated with excellent results in guiding ablation of typical AVNRT, the latter offers significantly shorter procedure and fluoroscopy times, improving the efficiency of the procedure and reducing X-ray exposure. PMID- 12633643 TI - Stability of far field R wave signals in different conditions. AB - AIMS: The presence of far field R wave sensing (FFRS) is usually evaluated in patients with dual chamber pacemakers in supine position. To check if this approach is valid, we tested whether FFRS is consistent both in terms of amplitude threshold and timing characteristics in different daily life conditions. METHODS AND RESULTS: In 42 patients with a DDD pacemaker, the presence, amplitude threshold and timing parameters of FFRS were therefore determined, with patients supine, standing and at peak exercise. Measurements were made of paced and sensed R waves, in unipolar and bipolar sensing configurations (at peak exercise only paced R waves and bipolar sensing). After paced R waves (bipolar sensing) amplitude thresholds/time of FFRS after V pace were 0.32+/-0.18 mV/119-139 ms (supine), 0.32+/-0.16 mV/114-130 ms (upright) and 0.27+/-0.13 mV/121-136 ms (exercise) - with unipolar sensing, this was 0.49+/ 0.27 mV/101-150 ms (supine), 0.51+/-0.29 mV/100-144 ms (upright). After sensed R waves (bipolar sensing) amplitude thresholds/time of FFRS after V sense were 0.27+/-0.18 mV/24-42 ms (supine), 0.29+/-0.16 mV/18 to 41 ms (upright) - with unipolar sensing, thresholds were 0.59+/-0.32 mV/3-50 ms (supine), 0.59+/-0.36 mV/2-58 ms (upright). CONCLUSION: given the lower FFRS thresholds with bipolar sensing, bipolar sensing is superior in avoiding FFRS compared with unipolar sensing. No differences were found in terms of amplitude thresholds and timing characteristics with patients supine, standing and at peak exercise. Thus, measurements made in the supine position are basically sufficient to predict the presence/absence of FFRS under different conditions. PMID- 12633644 TI - Unusual extraction of an active fixation ventricular pacing lead with outer coil fracture in a child. AB - AIM: In this report we describe an asymptomatic paced child with outer coil fracture of the bipolar screw-in steroid eluting ventricular lead without insulation defect, loss of ventricular capture and unexpected increase in lead impedance in the bipolar VVIR pace configuration. METHOD AND RESULT: A previously unpublished method was performed to retract the helix and the lead. As a result, the fractured lead was successfully retrieved. CONCLUSION: We suggest that, this unusual extraction method can be tried as an alternative approach in the removal of an active fixation ventricular pacing lead with impaired mechanical function possibly due to fracture. PMID- 12633645 TI - Pacing for adult patients with left atrial isomerism: efficacy and technical considerations. AB - AIM: Left atrial isomerism (LAI) is an uncommon congenital heart disease, which is combined with a range of intracardiac abnormalities. Atrial arrhythmia is common in LAI. We hypothesized that permanent pacing will decrease arrhythmia recurrence in adult patients with LAI. METHODS AND RESULTS: Patients with LAI and permanent pacing were identified from the Royal Brompton Adult Congenital Heart Database. Hospital records were reviewed, and patients' current status was assessed with clinic reviews including standard and Holter ECGs, and transthoracic echocardiograms. Four patients who underwent permanent pacing at the age of 27.5+/-11.7 years fulfilled inclusion criteria, namely permanent pacing in the setting of LAI. All 4 had sick sinus syndrome-with sustained supraventricular arrhythmia in 3-requiring several hospitalizations for arrhythmia management prior to pacing. All 4 patients remained free of clinical arrhythmia and hospital admission at a mean follow-up of 67.0+/-59.5 months from pacing. The latest ECG revealed paced rhythm in all. NYHA functional class remained stable. There were no pacemaker related complications at implantation or during follow-up. CONCLUSION: Adults with LAI benefit from pacemaker implantation, which reduces recurrence of supraventricular tachycardia and maintains atrioventricular synchrony. Future studies need to determine the preferred pacemaker mode and optimal timing for pacing. PMID- 12633646 TI - Successful percutaneous extraction of an inadvertently placed left ventricular pacing lead. AB - A 74-year-old patient was referred for a rapidly increasing pacing threshold 9 months after DDD pacemaker implantation because of symptomatic total atrioventricular (AV) block. She had a history of hypertension, diabetes with micro-angiopathy and a recent transient ischaemic attack. The paced electrocardiogram on admission had a right bundle branch block pattern and 3 dimensional transoesophageal echocardiography demonstrated passage of the lead through an atrial septal defect with a left ventricular position in addition to moderate atherosclerosis of the ascending aorta. No thrombus could be detected on the lead. Percutaneous extraction is usually not recommended because of the risk of mobilization of thrombus material. However, the risk of stroke during removal using cardiopulmonary bypass in this patient was considerably increased because of the presence of multiple independent risk factors. Therefore, percutaneous extraction using a locking device was selected and performed without complications: follow-up was uneventful. PMID- 12633647 TI - Antibradycardia pacing in patients with congenital heart disease: experience with automatic threshold determination and output regulation (Autocapture). AB - AIMS: The purpose of this study was to reevaluate whether St Jude Medical's Autocapture algorithm (AC) with beat-to-beat capture confirmation, automatic pacing threshold determination and output adjustment can be applied to paced patients with congenital heart disease (CHD). METHODS AND RESULTS: 30 patients with CHD received a transvenous (group A: n=22) or epicardial (group B: n=8) single (n=7) or dual chamber (n=23) AC pacemaker for antibradycardia pacing. As a safe AC function is ensured only if a sufficient evoked response (ER) and a low lead polarization amplitude (LPA) are present, these parameters were reevaluated from 112 follow-up studies with respect to AC-function. In all but one transvenous system AC correctly functioned. AC was recommended in 5/8 patients with epicardial leads but correct AC function was preserved in only 3 patients. CONCLUSION: These data suggest that the application of the AC algorithm is safe in patients with CHD when transvenous leads are used. Whether appropriate AC function is possible with epicardial leads needs individual verification. PMID- 12633648 TI - Short and long-term single-centre experience with an S-shaped unipolar lead for left ventricular pacing. AB - Left ventricular-based pacing is an established method for treatment of congestive heart failure in patients with ventricular dyssynchrony. The transvenous epicardial approach is the method of choice to pace the left ventricle. AIMS: To evaluate short and long-term stability and pacing and sensing performance of an S-shaped non-steroid unipolar lead. METHODS: Forty-eight procedures were performed in 43 consecutive patients (mean age: 70+/-8 years, 32 males) with severe congestive heart failure. The left ventricular lead was placed into a coronary sinus tributary. Pacing and sensing thresholds and pacing impedance were measured at implant, 1 and 6 months. RESULTS: The mean procedure time was 90.0+/-35.5 min. Pacing thresholds at implant, 1 and 6 months were 1.1+/ 0.8 V, 1.9+/-1.3 V and 1.9+/-1.5 V respectively. In 7 patients, lead implantation was unsuccessful. One of them had a successful second attempt. Lead revision was performed in 5 patients for loss of capture. CONCLUSION: The S-shaped unipolar lead evaluated in this study provides stable long-term position and pacing thresholds. Recent improvement of this S-shaped lead model will hopefully reduce the rate of implantation failures and acute dislodgements. PMID- 12633649 TI - Fish microsporidia: fine structural diversity and phylogeny. AB - Structural diversity of fish microsporidian life cycle stages and of the host parasite interface is reviewed. In the infected cell of the fish host, microsporidia may either cause serious degradation of the cytoplasm and demise of the cell, or they may elicit host cell hypertrophy, producing a parasite hypertrophic host cell complex, the xenoma. The structure of the xenoma and of its cell wall may differ according to the genus of the parasite, and seems to express properties of the parasite rather than those of the host. In merogony, the parasite cell surface interacts with the host cell in diverse ways, the most conspicuous being the production of thick envelopes of different types. Sporogony stages reveal different types of walls or membranes encasing the sporoblasts and later the spores and these envelopes may be of host or parasite origin. Nucleospora differs from all other fish microsporidia by its unique process of sporogony. Except for the formation of conspicuous xenomas, there are no essentially different structures in fish-infecting microsporidia compared with microsporidia from other hosts. Although the structures associated with the development of fish microsporidia cannot be attributed importance in tracing the phylogeny, they are relevant for practical determination and assessing the relation to the host. The possibility of the existence of an intermediate host is discussed. Higher-level classification of Microsporidia is briefly discussed and structure and evolutionary rates in microsporidian rDNA are reviewed. Discussion of rDNA molecular phylogeny of fish-infecting microsporidia is followed by classification of these parasites. Most form a rather cohesive clade. Outside this clade is the genus Nucleospora, separated at least at the level of Order. Within the main clade, however, there are six species infecting hosts other than fish. Based on data available for analysis, a tentative classification of fish infecting microsporidia into five groups is proposed. Morphologically defined groups represent families, others are referred to as clades. Group 1, represented by family Pleistophoridae, includes Pleistophora, Ovipleistophora and Heterosporis; Vavraia and Trachipleistophora infect non-fish hosts. Group 2, represented by family Glugeidae, is restricted to genus Glugea and Tuzetia weidneri from crustaceans. Group 3 comprises three clades: Loma and a hyperparasitic microsporidian from a myxosporean; Ichthyosporidium and Pseudoloma clade and the Loma acerinae clade. For the latter species a new genus has to be established. Group 4 contains two families, Spragueidae with the genus Spraguea and Tetramicridae with genera Microgemma and Tetramicra, and the Kabatana and Microsporidium seriolae clade. Group 5 is represented by the family Enterocytozoonidae with the genus Nucleospora and mammal-infecting genus Enterocytozoon. PMID- 12633651 TI - Parasite infectivity to hybridising host species: a link between hybrid resistance and allopolyploid speciation? AB - Variation in host-specific infectivity was studied in monogenean polystome parasites (Protopolystoma spp.) of the interfertile, parapatric anurans Xenopus laevis laevis and Xenopus muelleri. Laboratory-raised host F1 hybrids were resistant to parasites respectively specific to each parent taxon in nature. This resistance occurred against parasite isolates from both inside and outside a host hybrid/sympatric zone (and no isolate was compatible with the foreign host species under experimental conditions). Geographical Protopolystoma xenopodis isolates showed variable infectivity to a single full-sib group of their usual host, X. l. laevis, and strains with high or low infectivity to these sibs co occurred in spatially distant local areas (separated by 1,700 km). The host compatibility of P. xenopodis was also subject to host genotypexparasite genotype interactions. Refractoriness to some parasites or pathogens, as a consequence of hybridisation, may have conferred a selective advantage on the allopolyploid pathway by which most Xenopus spp. are believed to have evolved. PMID- 12633650 TI - Protein disulphide isomerase of Ostertagia ostertagi: an excretory-secretory product of L4 and adult worms? AB - A pepstatin A-agarose column was used in an attempt to purify a previously described antibody-degrading aspartyl proteinase from excretory-secretory material from the L4 and the adult stages of the bovine abomasal nematode Ostertagia ostertagi. However, no aspartyl proteinase activity was detected in the eluted fractions (L4Pepst and AdPepst). Screening of cDNA libraries with polyclonal antibodies raised against L4Pepst and AdPepst showed that a protein disulphide isomerase (Ost-PDI2) was present in both antigen fractions. This multifunctional enzyme was detected in extracts of L3, L4 and adult parasites and, interestingly, also in excretory-secretory material of L4 and adult O. ostertagi. By immunohistochemistry, the Ost-PDI2 enzyme was localised in some parts of the hypodermis of L4 and adult worms and in the intestinal cells of all three parasitic life stages. Two-dimensional Western blot analysis indicated that Ost-PDI2 is recognised by calves during a natural O. ostertagi infection, which suggests that Ost-PDI2 could be used for immunological control of ostertagiosis. PMID- 12633652 TI - Acclimation of entomopathogenic nematodes to novel temperatures: trehalose accumulation and the acquisition of thermotolerance. AB - The effect of thermal acclimation on trehalose accumulation and the acquisition of thermotolerance was studied in three species of entomopathogenic nematodes adapted to either cold or warm temperatures. All three Steinernema species accumulated trehalose when acclimated at either 5 or 35 degrees C, but the amount of trehalose accumulation differed by species and temperature. The trehalose content of the cold adapted Steinernema feltiae increased by 350 and 182%, of intermediate Steinernema carpocapsae by 146 and 122% and of warm adapted Steinernema riobrave by 30 and 87% over the initial level (18.25, 27.24 and 23.97 microg trehalose/mg dry weight, respectively) during acclimation at 5 and 35 degrees C, respectively. Warm and cold acclimation enhanced heat (40 degrees C for 8h) and freezing (-20 degrees C for 4h) tolerance of S. carpocapsae and the enhanced tolerance was positively correlated with the increased trehalose levels. Warm and cold acclimation also enhanced heat but not freezing tolerance of S. feltiae and the enhanced heat tolerance was positively correlated with the increased trehalose levels. In contrast, warm and cold acclimation enhanced the freezing but not heat tolerance of S. riobrave, and increased freezing tolerance of only warm acclimated S. riobrave was positively correlated with the increased trehalose levels. The effect of acclimation on maintenance of original virulence by either heat or freeze stressed nematodes against the wax moth Galleria mellonella larvae was temperature dependent and differed among species. During freezing stress, both cold and warm acclimated S. carpocapsae (84%) and during heat stress, only warm acclimated S. carpocapsae (95%) maintained significantly higher original virulence than the non-acclimated (36 and 47%, respectively) nematodes. Both cold and warm acclimated S. feltiae maintained significantly higher original virulence (69%) than the non-acclimated S. feltiae (0%) during heat but not freezing stress. In contrast, both warm and cold acclimated S. riobrave maintained significantly higher virulence (41%) than the non-acclimated (14%) nematodes during freezing, but not during heat stress. Our data indicate that trehalose accumulation is not only a cold associated phenomenon but is a general response of nematodes to thermal stress. However, the extent of enhanced thermal stress tolerance conferred by the accumulated trehalose differs with nematode species. PMID- 12633653 TI - Leishmania (Leishmania) chagasi infection alters the expression of cell adhesion and costimulatory molecules on human monocyte and macrophage. AB - The initial steps of Leishmania infection in humans are largely unknown. There is limited information on the Leishmania infected human monocytes, the first cells that the parasite lives in, particularly related to costimulatory molecules. We show here that Leishmania (L.) chagasi infection avoids inducing proinflammatory molecules and has striking down modulating effects on human monocytes or macrophages. It does not induce CD54, interleukin (IL)-12 or tumour necrosis factor-alpha, potent proinflammatory cytokines and down modulates CD11b expression in monocytes. Lipopolysaccharide stimulated IL-12 (p40) levels, CD54 and HLA-DR expression are diminished in infected monocytes as well as interferon gamma stimulated HLA-DR and HLA-ABC expression in infected macrophages. There is a negative correlation between CD54 and CD86 expression in both monocytes and macrophages. The depressed expression of class I and II molecules, absence of key proinflammatory cytokines and impaired expression of costimulatory molecules induced by L. chagasi could leave the immune system, at least in its initial phases in anergy or ignorance. PMID- 12633654 TI - Molecular dissection of the human B-cell response against Toxoplasma gondii infection by lambda display of cDNA libraries. AB - The disorders generated by Toxoplasma gondii infection are closely associated with the competence of the host immune system and both humoral and cell mediated immunity are involved in response to parasite invasion. To identify antigens implicated in human B-cell responses, we screened a phage-display library of T. gondii cDNA fragments with sera of infected individuals. This approach identified a panel of recombinant phage clones carrying B-cell epitopes. All the peptide sequences selected by this procedure are regions of T. gondii gene products. These regions contain epitopes of the T. gondii antigens SAG1, GRA1, GRA7, GRA8 and MIC5, which are recognised by human immunoglobulins. Moreover, we report the isolation and characterisation of two additional immunodominant regions encoded by GRA3 and MIC3 genes, whose products have never been described as antigens of the human B-cell response against T. gondii infection. These results demonstrate potential of lambda-display technology for antigen discovery and for the study of the human antibody response against infectious agents. PMID- 12633655 TI - Influence of CD4+CD25+ T cells on Plasmodium berghei NK65 infection in BALB/c mice. AB - CD4(+) T cells co-expressing CD25 (CD4(+)CD25(+) T cells) have been identified as immunoregulatory suppressors modulating autoimmune response. Beside that, autoimmune response was supposed to be associated with malaria infection. Based on these data, we hypothesised that CD4(+)CD25(+) T cells may influence protective immunity to malaria parasites, while suppressing autoimmune response arising throughout the course of malarial infection. To test this possibility, we evaluated the kinetics of CD4(+)CD25(+) T cells during malaria infection and investigated the influence of CD25 depletion by anti-mouse CD25 monoclonal antibody (PC61) on the infection, using a mouse model of premunition to Plasmodium berghei NK65 malaria. The results showed that, during exacerbation of P. berghei NK65 infection, the proportion of CD4(+)CD25(+) T cells among CD4(+) T cells decreased, although that of CD4(+) T cells increased. CD25 depletion clearly delayed the growth of parasitaemia during parasite challenge, particularly in immunised mice. These findings demonstrated that CD4(+)CD25(+) T cells are able to influence protective immunity underlying premunition to P. berghei NK65 parasites. PMID- 12633656 TI - Acidification modulates the traffic of Trypanosoma cruzi trypomastigotes in Vero cells harbouring Coxiella burnetii vacuoles. AB - We studied the fate of different Trypanosoma cruzi trypomastigote forms after they invade Vero cells persistently colonised with Coxiella burnetii. When the invasion step was examined we found that persistent C. burnetii infection per se reduced only tissue-culture trypomastigote invasion, whereas raising vacuolar pH with Bafilomycin A1 and related drugs, increased invasion of both metacyclic and tissue-culture trypomastigotes when compared with control Vero cells. Kinetic studies of trypomastigote transfer indicated that metacyclic trypomastigotes parasitophorous vacuoles are more efficiently fused to C. burnetii vacuoles. The higher tissue-culture trypomastigote hemolysin and transialidase activities appear to facilitate their faster escape from the parasitophorous vacuole. Sialic acid deficient Lec-2 cells facilitate the escape of both forms. Endosomal lysosomal sequential labelling with EEA1, LAMP-1, and Rab7 of the parasitophorous vacuoles formed during the entry of each infective form revealed that the phagosome maturation processes are also distinct. Measurements of C. burnetii vacuolar pH disclosed a marked preference for trypomastigote fusion with more acidic rickettsia vacuoles. Our results thus suggest that intravacuolar pH modulates the traffic of trypomastigote parasitophorous vacuoles in these doubly infected cells. PMID- 12633657 TI - Effects of KHEYLRFamide and KNEFIRFamide on cyclic adenosine monophosphate levels in Ascaris suum somatic muscle. AB - KHEYLRF-NH(2) (AF2) is a FMRFamide-related peptide (FaRP) present in parasitic and free-living nematodes. At concentrations as low as 10 pM, AF2 induces a biphasic tension response, consisting of a transient relaxation followed by profound excitation, in neuromuscular strips prepared from Ascaris suum. In the present study, the effects of AF2 on cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP) and inositol-1,4,5-triphosphate (IP(3)) levels were measured following muscle tension recordings from 2 cm neuromuscular strips prepared from adult A. suum. AF2 induced a concentration- and time dependent increase in cAMP, beginning at 1 nM; cAMP levels increased by 84-fold following 1 h exposure to 1 microM AF2. cGMP and IP(3) levels were unaffected by AF2 at concentrations 10 years; use of insulin; serum creatinine >2 mg/dl; urinary protein >300 mg per 24 hours; presence of peripheral vascular disease (ankle:brachial ratio <1.0); and documentation of other diabetic comorbidity (retinopathy, neuropathy, gastroparesis). RESULTS: During this time period, 374 adult cardiac transplants were performed. Seventy-six patients (20%) were diabetic with 33 patients (43%) requiring insulin. Forty-two of the patients had moderate disease. Survival of the diabetic and non-diabetic recipients was comparable (1- and 3-year survival of 86% and 85%. vs 87% and 84%, respectively, p = NS). No difference in survival between "medium-risk" and "low-risk" diabetics was observed. The incidence of acute rejection in the first year, graft vasculopathy and infection, was comparable between diabetic and non-diabetic patients. In both diabetic and non-diabetic patients, there was a similar and small insignificant increase in serum creatinine. CONCLUSIONS: More patients with advanced diabetes are undergoing cardiac transplantation and the early and mid term survival remains comparable to non-diabetic recipients. Future liberalization of transplantation in diabetics appears likely. PMID- 12633691 TI - Long-term results of CMV hyperimmune globulin prophylaxis in 377 heart transplant recipients. AB - BACKGROUND: Cytomegalovirus (CMV) has emerged as the most important pathogen to affect the post-operative course after heart transplantation. We performed a retrospective analysis to evaluate the efficiency of CMV hyperimmune globulin (CMVIG) prophylaxis in preventing CMV disease in aggressively immunosuppressed patients after heart transplantation. METHODS: We studied 377 heart transplant recipients who received quadruple-immunosuppressive therapy and CMVIG as sole CMV prophylaxis. The study population was categorized into 4 groups according to donor and recipient CMV serology at the time of transplantation (D+/R+, D+/R-, D /R+, D-/R-) and was monitored for CMV immediate early antigen in peripheral blood cells, in urine sediments, and in throat washings; for the presence of serum CMV immunoglobulin M and CMV immunoglobulin G; and for clinical evidence of CMV related symptoms. In addition, we compared the incidence of cardiac allograft vasculopathy and infection among the groups. RESULTS: During the first 5 years after transplantation, CMV disease developed in 79 patients (20.96%). Comparison among the groups showed significantly increased risk for CMV disease in allograft recipients of organs from seropositive donors (D+, 27.31%; D-, 11.33%; p = 0.0003). We observed 6 CMV-associated deaths, all in CMV-antibody-negative recipients. Additionally CMV-positive recipients had a greater incidence of cardiac allograft vasculopathy (p = 0.048), and a greater overall infection rate (p = 0.0034). CONCLUSIONS: Cytomegalovirus hyperimmune globulin administration prevents CMV disease and infection in aggressively immunosuppressed heart transplant recipients. Because fatal CMV disease in CMV-negative recipients of organs from seropositive donors could not be prevented with CMVIG alone, we recommend the additional use of prophylactic ganciclovir in this CMV-mismatched population. PMID- 12633692 TI - Aspergillus infections after lung transplantation: clinical differences in type of transplant and implications for management. AB - BACKGROUND: Invasive aspergillosis is a serious opportunistic infection in lung transplant recipients. It has not been fully discerned whether there are differences in the characteristics, risk factors and outcome of Aspergillus infection in single as compared with bilateral lung transplant recipients. METHODS: English-language articles identified by a MEDLINE search through December 2000 and bibliographies were used as data sources to identify cases of Aspergillus infections in lung transplant recipients. The studies selected had to have provided a definition of invasive aspergillosis to distinguish colonization from infection. RESULTS: The median incidence of Aspergillus infections in lung transplant recipients was 6.2%. In total, 58% (45 of 78) of the Aspergillus infections were tracheobronchitis or bronchial anastomotic infections, 32% (25 of 78) were invasive pulmonary, and 22% (25 of 78) were disseminated infections. Single lung transplant recipients with Aspergillus infections were significantly older (p = 0.006), more likely to have had chronic obstructive pulmonary disease as an underlying illness (p = 0.05), more likely to have developed Aspergillus infections later after transplantation (p = 0.019), and tended to have a higher incidence of invasive aspergillosis (p = 0.11) than all other lung transplant recipients. Overall mortality in lung transplant recipients with Aspergillus infections was 52%. Single lung transplant recipients (p = 0.03), and patients with late-onset infections (occurring at least 3 months after transplantation ([p = 0.045]) infections had significantly higher mortality. CONCLUSIONS: Single lung transplant recipients with Aspergillus infections had an overall greater morbidity and poorer outcome than other types of lung transplant recipients. Recognition of the unique characteristics of Aspergillus infections in single lung (vs bilateral or heart-lung) transplant recipients has implications relevant for the management of lung transplant recipients with aspergillosis. PMID- 12633693 TI - Diagnostic properties of transbronchial biopsy in lung transplant recipients who require mechanical ventilation. AB - INTRODUCTION: Bronchoscopy with transbronchial biopsy (TBBx) and bronchoalveolar lavage is useful and safe for diagnosing acute rejection and infection in lung transplant recipients. However, its role is less well defined in determining the etiology of allograft dysfunction in the setting of respiratory failure necessitating mechanical ventilation. METHODS: We retrospectively identified 41 mechanically ventilated patients with respiratory failure in whom 42 TBBx were followed within a 10 day period by surgical lung biopsy (SLBx) to determine the sensitivity, specificity, and positive and negative predictive values of TBBx compared with SLBx. RESULTS: The sensitivity, specificity, and positive and negative predictive values of TBBx for all episodes of acute rejection and for significant episodes of acute cellular rejection were 53.3% and 36.0%; 91.7% and 94.1%; 94.1% and 90.0%; 44.0% and 50.0%, respectively. A significantly higher histologic grade was noted on SLBx compared with TBBx specimens obtained within a 10-day period (2.39 +/- 1.02 vs 0.97 +/- 0.11, p 1 year; and (3). with a relatively lower cyclosporine level. CONCLUSIONS: These data suggest that continued surveillance of pediatric transplant patients for acute rejection is indicated for long-term follow-up. PMID- 12633695 TI - Low incidence and severity of transplant-associated coronary artery disease in heart transplants from live donors. AB - BACKGROUND: Hearts transplanted from patients undergoing heart-lung transplantation (domino hearts) are unique because they have not been subjected to the deleterious effects of brain-stem death. This study examines the incidence and severity of transplant-associated coronary artery disease in recipients of domino hearts. METHODS: We retrospectively reviewed angiographic and clinical data from 97 patients who survived more than 1 year after domino heart transplantation at our hospital. Duration of follow-up ranged from 1 to 11 years after transplantation. The diagnosis of coronary artery disease was based on angiographic criteria. RESULTS: At 1 year, freedom from angiographic coronary artery disease was 99% (70% confidence interval [CI], 97-100), at 5 years it was 83% (70% CI, 78-89), and at 10 years it was 77% (70% CI, 70-84). Donor age, cystic fibrosis in the donor, organ ischemia time during transplantation, and acute rejection after transplantation did not influence risk for the disease. We found an increased incidence of coronary disease in hearts from male donors compared with those from female donors: freedom from disease at 5 years was 72% (70% CI, 63-81) in men vs 93% (87-99) in women. Thirteen patients experienced coronary artery disease at a median of 3 years after transplantation; 4 patients died but most patients remained asymptomatic with angiographically mild disease at their last follow-up examination. CONCLUSIONS: We found decreased incidence and severity of transplant-associated coronary artery disease in recipients of domino hearts compared with that reported in recipients of cadaveric hearts. This data supports the continued practice of domino heart transplantation and also supports the hypothesis that brain death may contribute to the development of transplant coronary artery disease in recipients of hearts transplanted from cadaveric organ donors. PMID- 12633696 TI - Cytomegalovirus antibody status of donor/recipient does not influence the incidence of bronchiolitis obliterans syndrome in lung transplantation. AB - BACKGROUND: We have previously reported that prophylaxis for cytomegalovirus (CMV) infection does not influence the incidence of bronchiolitis obliterans syndrome (BOS) at 2 years. The effect of CMV infection (without evidence of disease) on BOS is still not well understood. Moreover, the incidence and risk factors for development of BOS in CMV-antibody-negative donor/recipient matches in lung transplantation have not been described. The aim of this study is to determine the incidence of BOS in lung transplant patients with CMV-antibody negative (-) donors (D) and recipients (R), and to evaluate the risk factors that predispose to BOS in this sub-group. METHOD: A retrospective study of data from the transplant database of our center was performed. All single-lung (SL), double lung (DL) and heart-lung block (HL) transplant patients who survived >2 years post-transplant were included in the study group. They were grouped as follows: D(-)/R(-), n = 102; D(-)/R(+), n = 70; D(+)/R(-), n = 33, and D(+)/R(+), n = 92. RESULTS: The 3-year BOS-free survival rates were 65%, 56%, 58% and 67%, respectively, and the incidence rates of BOS at 5 years post-transplant in the different groups were 57%, 62%, 78% and 55% (p > 0.05). In the D(-)/R(-) group, the significant risk factor for developing BOS was three or more episodes of acute rejection (p = 0.02). The mean numbers of acute rejection episodes per 100 patients-days within the first 6 months were 1.28, 1.06, 0.50 and 1.11 (p < 0.001 overall) for the four groups, respectively. CONCLUSION: Although CMV is believed to be a risk factor for BOS, its absence did not affect the occurrence or incidence of BOS in lung transplant patients. The main risk factor for BOS in the CMV-antibody-negative population remains the number of acute rejection episodes within the first 6 months after transplantation. PMID- 12633697 TI - Left ventricular function during support with an asynchronous pulsatile left ventricular assist device. AB - BACKGROUND: Left ventricular assist devices (LVADs) are frequently used to maintain patients with severe heart failure until heart transplantation becomes possible. Some patients may experience recovery of LV function during such support. Therefore, it is essential to be able to monitor changes in LV function in this setting. METHODS: We studied LV function in 10 patients (median age 34 years, 9 male) who had LVADs implanted because of severe heart failure due to dilated cardiomyopathy a median of 4 months previously. Median pre-implant ejection fraction was 27% and all patients had been on maximal medical therapy, including intravenous inotropic support, prior to insertion of the LVAD. RESULTS: During LVAD support there were cyclical variations in LV dimensions, fractional shortening (FS) and transmitral flow, related to changes in the phase relationship of the LV and the LVAD. The "best" FS occurred when LV systole coincided with device filling and the "worst" FS when LV systole coincided with device ejection. Median FS with the pump switched off was 18% (10% to 32%). Pump off FS was significantly greater than the "worst" FS with the pump on (5%, p = 0.002), and similar to the "best" pump-on FS (19%, p = NS). CONCLUSIONS: LV function could be studied echocardiographically during LV support and brief periods of interruption in support. Function varied according to the phase relationship of the LV and LVAD. The "best" FS measured during LVAD support was more closely related to the FS with the device switched off than the "worst" pump on FS. The "best" pump-on LV function is therefore most representative of intrinsic LV performance and can be used as a guide to recovery and the potential need for pump-off studies. PMID- 12633698 TI - Partial left ventriculectomy in modified adriamycin-induced cardiomyopathy in the dog. AB - BACKGROUND: The purpose of this study was to evaluate modified adriamycin-induced cardiomyopathy in the dog for research on partial left ventriculectomy (PLV). METHODS: An intracoronary catheter was introduced into the left main stem via the first marginal branch in a retrograde fashion in 12 adult foxhound dogs. The catheter was connected to a percutaneous access port that was used for weekly adriamycin administration (10 mg over a 1-hour period on 5 occasions). Follow-up examinations (transthoracic echocardiography, hemodynamic parameters, cardiopulmonary status, neurohormones) were done before, 1 week after the last adriamycin administration, and then 6 weeks later. This protocol was performed in 6 dogs (control group: Group 1). The other 6 dogs underwent PLV 1 week after the last adriamycin administration (Group 2). After the last measurements, all dogs were killed with saturated potassium chloride under general anesthesia and the hearts were excised for histologic examination. All data were calculated as mean and standard error of the mean. Differences were calculated by the Wilcoxon signed-rank test for paired and unpaired data. p < 0.05 was considered statistically significant. RESULTS: One dog from each group died suddenly during adriamycin administration (probably due to ventricular arrhythmia). In addition, 1 dog from Group 2 suffered from a severe systemic inflammatory response syndrome after PLV and died 36 hours after surgery. Thus, 5 dogs from Group 1 and 4 from Group 2 underwent the entire study protocol. Adriamycin administration resulted in a severe dilated cardiomyopathy that was comparable in both groups (significant increase of central venous pressure, mean pulmonary artery pressure, pulmonary wedge pressure, left ventricular end-systolic and end-diastolic diameters, oxygen extraction, troponin I and anti-diuretic hormone, whereas cardiac output, ejection fraction and venous oxygen saturation decreased significantly). Deterioration of cardiac function continued after termination of adriamycin administration in Group 1 dogs, albeit not as progressively as during adriamycin administration. In contrast, cardiac function improved in Group 2 dogs after PLV, but did not reach baseline values. Cardiac index increased and oxygen extraction (p = 0.03) decreased, resulting in an enhanced venous oxygen saturation (p = 0.02). In particular, the distance of the papillary muscles at end diastole (p = 0.02) and at end systole (p = 0.02) at the mid-papillary level decreased significantly after PLV, resulting in reduced left ventricular diameter and volume (statistically significant for left ventricular end-systolic diameter and volume). All hearts had severe histologic alterations characteristic of adriamycin-induced toxicity, including cytoplasmic vacuolation, myocyte degeneration and increased fibrosis. CONCLUSION: Modified adriamycin-induced cardiomyopathy in the dog may be suitable for research on PLV. PMID- 12633699 TI - Lifestyle and quality of life in long-term cardiac transplant recipients. AB - BACKGROUND: The purposes of this study were to describe long-term (>1 year) cardiac transplant recipients' perceptions of barriers to health-promoting behaviors; ability to manage their health, health-promoting lifestyle, health status and quality of life (QOL); and determine predictors of QOL. METHODS: Data were collected from 93 recipients who completed a self-report questionnaire incorporating: (1). patient characteristics; (2). barriers to health promotion, perceived health competence and health-promoting lifestyle; (3). perceived health status; and (4). QOL. All instruments used had acceptable reliability and validity. Objective measures of health status were retrieved from medical records. Data were analyzed using descriptive and comparative statistics, measures of association and multiple regression. RESULTS: Recipients were uncertain about their ability to manage their health; felt that barriers sometimes interfered with engaging in health-promoting behaviors; and often included health-promoting behaviors in their lifestyle. They rated their health as good and were moderately satisfied with life. Predictors of better perceptions of QOL included less education, longer time since transplant, ischemic etiology of heart failure, fewer barriers, higher perceived health competence and a health promoting lifestyle (R(2) = 0.51; F = 14.77; p = 0.001). CONCLUSIONS: The present results provide information regarding factors that contribute to perceptions of quality of life after cardiac transplantation, have implications for identifying patients at risk for poor QOL, and sensitize clinicians to the need for ongoing patient education. PMID- 12633700 TI - Change in quality of life from before to after discharge following left ventricular assist device implantation. AB - BACKGROUND: Quality of life (QOL) outcomes after left ventricular assist device (LVAD) implantation from before to after hospital discharge have been examined only in a very small sample of patients. The purposes of this study are to describe change in QOL from before to after hospital discharge in LVAD patients and to determine whether being discharged with an LVAD predicts better QOL than being hospitalized with an LVAD. METHODS: A non-random sample of 62 LVAD patients (approximately 50 years old, male, white, married, fairly well-educated) completed self-report questionnaires at >or=2 timepoints post-implant. The questionnaires (Quality of Life Index, Rating Question Form, Heart Failure Symptom Checklist, Sickness Impact Profile, LVAD Stressor Scale, Jalowiec Coping Scale), which were collated into booklets, had acceptable reliability and validity. Longitudinal analyses were performed in 2 steps using 1-sample t-tests and linear mixed effects modeling. RESULTS: Perception of QOL and health status were fairly good both before and after discharge of LVAD patients. Discharge predicted increased satisfaction with socioeconomic areas of life; decreased overall and psychologic stress and stress related to family and friends, self care and work/school/finances; and decreased physical and self-care disability. CONCLUSIONS: QOL outcomes improved from before to after hospital discharge in LVAD patients awaiting heart transplantation. As LVADs potentially become available as destination therapy, in addition to being successful bridges to heart transplantation, QOL outcomes will become more important to study. PMID- 12633701 TI - The impact of left ventricular dysfunction on cardiac donor transplant rates. AB - BACKGROUND: Because of the shortage of heart donors in the United States, efforts are necessary to maximize the yield of donor screening. The purpose of this study was to quantify the effects of left ventricular (LV) systolic dysfunction on heart donor use. METHODS: Using the California Transplant Donor Network database, the records of all potential organ donors screened between January 1997 and June 1998 were reviewed. After excluding subjects for whom family consent could not be obtained and subjects <13 or >or=60 years old, a study group of 223 potential heart donors was analyzed. The number of hearts not used because of LV dysfunction, coronary artery disease (CAD), valvular disease, and LV hypertrophy were quantified. A logistic regression model was developed to quantify the independent effect of LV dysfunction on donor use rates after adjustment for age, weight, and cause of death. RESULTS: Ninety-nine (44%) of the 223 potential donor hearts were not transplanted. Thirty-six of these hearts were not transplanted because of cardiac causes, primarily LV dysfunction (26 cases) and CAD (8 cases). The multivariable analysis showed that after adjusting for other donor variables, ejection fraction was the most significant predictor of non-use, with an odds ratio of 1.48 per 5-point decrease in ejection fraction. CONCLUSIONS: Left ventricular dysfunction is an important cause of failure to transplant adult donor hearts. Efforts to improve the yield of heart donor screening should focus on prevention or reversal of LV dysfunction. PMID- 12633702 TI - Ischemia-reperfusion decreases protein tyrosine phosphorylation and p38 mitogen activated protein kinase phosphorylation in rat lung transplants. AB - BACKGROUND: Dramatic alterations of protein tyrosine phosphorylation have been found during the ischemia-reperfusion (IR) period of human lung transplantation. IR also induces activation of p38 mitogen-activated protein kinase (p38) in the heart and kidney. The objective of the present study was to determine whether these changes exist in a rat single-lung transplant model for further mechanistic investigations. METHODS: Isogeneic lung transplantation was performed from Lewis (LEW) to LEW rats, whereas allogeneic transplantation was from LEW to Brown Norway (BN) rats. Blood gases and peak airway pressure were monitored. Lung tissues were collected after 6 hours of cold ischemic preservation, after 30 minutes of warm ischemia for lung implantation, and after 2 hours of reperfusion. Protein tyrosine kinase (PTK) and phosphatase (PTP) activities were measured. Protein tyrosine phosphorylation, Src PTK and p38 expression and p38 phosphorylation were examined by western blotting. RESULTS: In both iso- and allografts, the lung function of transplants was very well preserved. Protein tyrosine phosphorylation, PTK and PTP activities were decreased significantly after 2 hours of reperfusion. Src protein level and phosphorylation of p38 were reduced after 2 hours of reperfusion. CONCLUSIONS: During the early IR period of lung transplantation, decreased protein tyrosine phosphorylation may be involved in apoptosis and other biologic changes. The lack of p38 activation suggests that activity of mitogen-activated protein kinase pathways in the lung transplantation setting may be different from other IR processes. PMID- 12633703 TI - Lazaroid (U74389G)-supplemented cardioplegia: results of a double-blind, randomized, controlled trial in a porcine model of orthotopic heart transplantation. AB - BACKGROUND: U74389G (16-desmethyl tirilazad), a 21-aminosteroid or "lazaroid," inhibits lipid peroxidation, which is an important element of ischemia reperfusion injury. The aim of this study was to determine whether the addition of U74389G to the cardioplegic preservation solution could improve early cardiac allograft function. METHODS: A porcine model of donor brain death and orthotopic cardiac transplantation was used. Hearts were arrested and preserved for 6 hours in an aspartate-enriched extracellular cardioplegia that had been supplemented with either U74389G and its carrier (n = 7) or the carrier alone (n = 9). Epicardial sonomicrometry and transmyocardial micromanometry were used to obtain pressure-volume loops before and after transplantation. Left ventricular wall volume was measured by volume displacement. RESULTS: A higher proportion of U74389G-treated hearts were weaned successfully from cardiopulmonary bypass, but this difference did not achieve statistical significance (86% [6 of 7] vs 56% [5 of 9]; p = 0.308). In the hearts that were weaned successfully, preservation of left ventricular contractility, as judged by the pre-load recruitable stroke work relationship, was significantly better in the U74389G-treated hearts (p = 0.0271). In contrast, left ventricular compliance, as judged by the end-diastolic pressure-volume relationship, was significantly better preserved in the control group (p < 0.0001). U74389G-treated hearts developed less myocardial edema, as judged by the post-transplant left ventricular wall volume/baseline steady-state epicardial end-diastolic volume ratio (64 +/- 9% vs 76 +/- 11%; p = 0.045). CONCLUSIONS: The benefit obtained from U74389G-supplemented cardioplegic preservation solution was marginal for hearts stored for 6 hours. After longer ischemic times, the benefit may be clearer. PMID- 12633704 TI - Recipient cells form the proliferative lesion in the rat heterotopic tracheal allograft model of obliterative airway disease. AB - To determine the nature of the proliferative lesion in obliterative airway disease, heterotopic tracheal allograft transplantation was performed between fully disparate Brown Norway and Lewis rat strains. Four weeks after transplantation, the resulting lumenal occlusive lesion was stripped from the underlying tissue. The lesion was probed using immunohistochemical analysis with monoclonal antibodies and for DNA using strain-specific primers for Brown Norway or Lewis major histocompatibility complex Class I alleles. The lesions were alpha actin positive, and polymerase chain reaction probing revealed only recipient DNA in the lesion tissue, regardless of the direction of transplantation, with no amplification of donor DNA. From this, we conclude that the proliferative lesion in the rat heterotopic tracheal model is of recipient origin a finding with important implications for the pathobiology of obliterative airway disease. PMID- 12633705 TI - Beta-blocker treatment before high-risk cardiac surgery. PMID- 12633706 TI - Fourier transform Raman spectroscopy of Syncrude Sweet Blend distillation fractions: the 200-1800 cm-1 region. AB - The fingerprint (200-1800 cm(-1)) region in FT-Raman spectra of Syncrude Sweet Blend (SSB) and its three constituent distillation fractions (naphtha, light gas oil and heavy gas oil) was analyzed in detail in this study. Approximately 50 bands were observed and assigned to functional groups in saturated (alkanes) and unsaturated (aromatics) species. Characteristic bands for mono-, bi-, and tricyclic aromatics were identified and used to quantify these groups in SSB and the distillation fractions. Total aromatics content was determined using the carbon-carbon stretching bands in the 1600-cm(-1) region, and shown to agree with earlier results obtained from the C-H stretching region. The bands due to mono- and bicyclic aromatics permitted calculation of the relative abundances of these species with an accuracy equivalent to that obtained using NMR spectroscopy, a traditional method for measuring this quantity. PMID- 12633707 TI - Phosphorescence spectra and triplet state lifetimes of palladium octaethylporphyrin, palladium octaethylchlorin and palladium 2,3 dimethyloctaethylisobacteriochlorin at 77 K. AB - The phosphorescence spectra and triplet state lifetimes of palladium octaethylporphyrin (PdOEP), palladium octaethylchlorin (PdOEC) and palladium 2,3 dimethyloctaethylisobacteriochlorin (PdOEiBC) in n-octane Shpolskii matrices at 77 K are reported. The lifetime and T(1)/S(0) origin energy of each complex are: PdOEP, 1.90+/-0.04 ms, 15162 cm(-1); PdOEC, 0.43+/-0.03 ms, 12547 cm(-1) and PdOEiBC, 0.59+/-0.03 ms, 12863 cm(-1). PMID- 12633708 TI - Spectrofluorimetric study of host-guest complexation of ibuprofen with beta cyclodextrin and its analytical application. AB - The characteristics of host-guest complexation between beta-cyclodextrin (beta CD) and two forms of ibuprofen (protonated and deprotonated) were investigated by fluorescence spectrometry. 1:1 stoichiometries for both complexes were established and their association constants at different temperatures were calculated by applying a non-linear regression method to the change in the fluorescence of ibuprofen that brought about by the presence of beta-CD. The thermodynamic parameters (deltaH, deltaS and deltaG) associated with the inclusion process were also determined. Based on the obtained results, a sensitive spectrofluorimetric method for the determination of ibuprofen was developed with a linear range of 0.1-2 microg ml(-1) and a detection limit of 0.03 microg ml(-1). The method was applied satisfactorily to the determination of ibuprofen in pharmaceutical preparations. PMID- 12633709 TI - Optical properties of Sm3+ ions doped in tellurite glass. AB - Judd-Ofelt analysis of Sm(3+) ions doped in a tellurite glass has been accomplished. Fluorescence and life-time measurements have been carried out using Ar(+) laser and excimer laser. Stark splitting has been observed in the upper and lower levels in fluorescence. The life-time of 4G(5/2) level as a function of Sm(3+) concentration has been measured. Concentration quenching and the mechanism responsible for the same have been found. PMID- 12633710 TI - Spectroscopic and optical studies on pure and doped single crystals of sulphate mixed L-arginine phosphate monohydrate--a nonlinear optical crystal. AB - Single crystals of pure and transition metal ions (Cu(2+) and VO(2+))-doped L arginine sulpho phosphate monohydrate (sulphate-mixed L-arginine phosphate monohydrate, abbreviated as LASP) have been grown by solvent evaporation of the saturated aqueous solution at room temperature and characterized by single crystal XRD, FT-IR, UV-Vis-NIR and EPR (single-crystal rotation) spectral studies. Kurtz powder technique shows an enhanced second harmonic generation (SHG) efficiency for LASP and its doped analogues than pure LAP. The EPR studies on LASP:VO(2+) reveal that the in-plane sigma-bonding is moderately covalent and the out-of-plane pi-bonding is highly covalent, which could be attributed to the cause of enhanced powder efficiency. PMID- 12633711 TI - Vibrational frequencies and structural determinations of tetraisocyanatosilane. AB - The normal mode frequencies and corresponding vibrational assignments of Si(NCO)(4) are examined theoretically using the GAUSSIAN 98 set of quantum chemistry codes. Each of the vibrational modes was assigned to one of six types of motion predicted by a group theoretical analysis (Si-N stretch, N-C-O symmetric stretch, N-C-O asymmetric stretch, N-C-O bend, Si-N-C bend, and N-Si-N bend) utilizing the T(d) symmetry of the molecule. Uniform scaling factors were derived for each type of motion. Predicted infrared and Raman intensities are reported. PMID- 12633712 TI - A temperature-dependent interaction of neutral red with calf thymus DNA. AB - Neutral red (NR) is used as a probe to study the temperature and concentration dependent interaction of a cationic dye with nucleic acid. A temperature dependent interaction of NR with calf thymus DNA (CT DNA) has been studied by differential pulse voltammetry (DPV), UV-Visible absorption, circular dichroism (CD) and fluorescence spectroscopy. The experimental results of increasing peak current, changes in the UV-Visible absorption and fluorescence spectra of NR and decreasing the induced circular dichroism (ICD) intensity show that (i) the binding mode of NR molecules is changed from intercalating into DNA base pairs to aggregating along the DNA double helix and (ii) the orientation of NR chromophore in DNA double helix is also changed with the temperature. PMID- 12633713 TI - Site-selective nitrogen isotopic ratio measurement of nitrous oxide using 2 microm diode lasers. AB - We demonstrate a high-precision measurement of the isotopomer abundance ratio 14N(15)N(16)O/15N(14)N(16)O/14N(14)N(16)O (approximately 0.37/0.37/100) using three wavelength-modulated 2 microm diode lasers combined with a multipass cell which provides different optical pathlengths of 100 and 1 m to compensate the large abundance difference. A set of absorption lines for which the absorbances have almost the same temperature dependence are selected so that the effect of a change in gas temperature is minimized. The test experiment using pure nearly natural-abundance N(2)O samples showed that the site-selective 15N/14N ratios can be measured relative to a reference material with a precision of +/-3 x 10(-4) (+/-0.3 per thousand) in approximately 2 h. PMID- 12633714 TI - Polarised vibrational spectra of betaine ortho-phosphoric acid complex. Part II. Phase transitions studies. AB - Polarised infrared transmission and Raman spectra of betaine ortho-phosphoric acid crystal in temperature ranges 13-393 and 13-300 K, respectively are reported and discussed in relation to phase transitions: antiferrodistortive at T(c1) = 365 K and antiferroelectric at T(c3) = 81 K. The spectra are consistent with unit cell doubling below T(c3). The participation of all hydrogen bonds apparent in the crystal in the antiferroelectric phase transition was shown. Quite large freedom of -N(CH(3))(3) groups reorientation in the antiferroelectric phase was detected. No changes were found in the transmission spectra taken in the vicinity of the antiferrodistortive phase transition temperature. PMID- 12633715 TI - Spectra and structure of silicon containing compounds. XXXII. Raman and infrared spectra, conformational stability, vibrational assignment and ab initio calculations of n-propylsilane-d0 and Si-d3. AB - The infrared (3100-40 cm(-1)) and Raman (3100-20 cm(-1)) spectra of gaseous and solid n-propylsilane, CH(3)CH(2)CH(2)SiH(3) and the Si-d(3) isotopomer, CH(3)CH(2)CH(2)SiD(3), have been recorded. Additionally, the Raman spectra of the liquids have been recorded and qualitative depolarization values obtained. Both the anti and gauche conformers have been identified in the fluid phases but only the anti conformer remains in the solid. Variable temperature (-105 to -150 degrees C) studies of the infrared spectra of n-propylsilane dissolved in liquid krypton have been recorded and the enthalpy difference has been determined to be 220+/-22 cm(-1) (2.63+/-0.26 kJ mol(-1)) with the anti conformer the more stable form. A similar value of 234+/-23 cm(-1) (2.80+/-0.28 kJ mol(-1)) was obtained for deltaH for the Si-d(3) isotopomer. At ambient temperature it is estimated that there is 30+/-2% of the gauche conformer present. The potential function governing the conformation interchange has been estimated from the far infrared spectral data, the enthalpy difference, and the dihedral angle of the gauche conformer, which is compared to the one predicted from ab initio MP2/6-31G(d) calculations. The barriers to conformational interchange are: 942, 970 and 716 cm(-1) for the anti to gauche, gauche to gauche, and gauche to anti conformers, respectively. Relatively complete vibrational assignments are proposed for both the n-propylsilane-d(0) and Si-d(3) molecules based on the relative infrared and Raman spectral intensities, infrared band contours, depolarization ratios, and normal coordinate calculations. The geometrical parameters, harmonic force constants, vibrational frequencies, infrared intensities, Raman activities and depolarization ratios, and energy differences have been obtained for the anti and gauche conformers from ab initio MP2/6-31G(d) calculations. Structural parameters and energy differences have also been obtained utilizing the larger 6-311 + G(d,p) and 6-311 + G(2d,2p) basis sets. From the isolated Si-H stretching frequency from the Si-d(2) isotopomer the r(0) distances of 1.484 and 1.485 A have been determined for the SiH(s) and SiH(a) bonds, respectively, for the anti conformer, and 1.486 A for the SiH bond for the gauche conformer. Utilizing previously reported microwave rotational constants for the anti conformer and the determined SiH distances along with ab initio predicted parameters 'adjusted r(0)' parameters have been obtained for the anti conformer. The results are discussed and compared to those obtained for some similar molecules. PMID- 12633716 TI - Calculation of the Raman frequencies of the translational mode in ammonia solid II. AB - We report here our calculated Raman frequencies of the translational mode as a function of temperature for the fixed pressures of 3.65, 5.02 and 6.57 kbars in the ammonia solid II. They were calculated by means of our Gruneisen relation using the volume data from the literature for all the pressures indicated within the temperature regions close to the melting point in this system. Our calculated frequencies are in very good agreement with those observed experimentally for this translational mode of the ammonia solid II. This shows that the observed behaviour of ammonia solid II can be described adequately by means of the calculation employed here. PMID- 12633717 TI - Molecular complexes of some anthraquinone anti-cancer drugs: experimental and computational study. AB - It is known that anti-cancer drugs target DNA in the cell. The mechanism of interaction of anti-cancer drugs with DNA is not fully understood. It is thought that the forces of interaction have some contribution from charge-transfer (CT) binding. The ability of some anthraquinones (AQs) anti-cancer drugs to form CT complexes with well-known electron donor molecules was investigated by NMR. The NMR spectroscopy has indicated the formation of CT complexes between 1,4-bis[[2 (dimethylamino) ethyl]amino]-5,8-dihydroxyanthracene-9,10-dione, (AQ4), and its des-hydroxylated equivalent 1,4-bis[[2-(dimethylamino) ethyl]amino]anthracene 9,10-dione, (AQ4H), as electron acceptors and pyrene (PY) and hexamethylbenzene (HMB) as electron donors. Association constants of the formed CT complexes were determined from the NMR data. AQ4 showed weaker electron accepting power than AQ4H, which could be easily explained on the basis of the electron donating nature of the two-hydroxyl groups. AQ4 and AQ4H have higher stability constant with PY than with HMB. This reflects the weaker interaction of the AQs with the latter, which is a direct effect of the six bulky methyl groups. Electronic absorption spectroscopy of the studied system was performed in chloroform and showed the absence of new absorption bands. The extent of interaction between AQs and donors has been computed using molecular mechanics and quantum mechanics. The computed values were compared with the experimental results of association constants. PMID- 12633718 TI - Application of spectroscopic and computational methods in the study of the electronic excitations of partially silylated hydroxyquinones. AB - The silylation of hydroxynaphthoquinones and hydroxyanthraquinones with N-methyl N-(tert-butyldimethylsilyl)-2,2,2-trifluoroacetamide, in one step reaction, under mild conditions, has led, in nearly quantitative yields, to the isolation of hydrolytically stable partially silylated products (only of the hydroxyl groups), which differ dramatically in color from the starting materials. The spectroscopic investigation of the silylated products in solution as well as the correlation of the observed data with computationally derived results have been carried out within a study aiming at the understanding of the visible maxima shift to either the ultraviolet or near-infrared region of the electromagnetic spectrum, which would imply availability of the silylated products in a variety of related applications. PMID- 12633719 TI - Solid-phase spectrophotometric determination of trace amounts of vanadium using 2,3-dichloro-6(3-carboxy-2- hydroxynaphthylazo)quinoxaline. AB - Solid-phase spectrophotometry (SPS) has been applied to analysis for trace amounts of vanadium in several environmental water (potable and polluted), biological samples (human blood and urine), and soil samples. Vanadium was sorbed in a styrene-divinylbenzene-type anion-exchanger Dowex 1-X8 as a vanadium--2,3 dichloro-6(3-carboxy-2-hydroxynaphthylazo)quinoxaline. Resin phase absorbances at 606 and 800 nm were measured directly which allowed the determination of vanadium in the range 0.03-2.2 ng ml(-1) with a relative standard deviation (R.S.D.) of 1.4%. The comparison of the SPS method and the gallic acid persulphate method shows that the linearity, analytical sensitivity, and precision were better for the SPS method, and that the latter method has lower detection and quantification limits compared with the gallic acid persulphate method. PMID- 12633720 TI - Optical gain characteristics of C 460 and C 450. AB - Dye concentration dependent gain spectra for Coumarin 460 (C 460) and Coumarin 450 (C 450) in ethanol have been studied using Amplified Spontaneous Emission (ASE) technique under Nitrogen laser (337.1 nm) excitation in the concentration range 10(-2)-10(-5) m/l. The dependence of lasing wavelength and peak gain on concentration have been understood in terms of variation of fluorescence lifetime, which is due to photo-physical processes such as radiation trapping and concentration-quenching. Pump intensity dependence of efficiency is also explained in terms of fluorescence lifetime. A comparison of the stability of the two dyes has also been made on the basis of the functional groups at different positions of the basic coumarin. PMID- 12633721 TI - Molecular recognition of 7-(2-octadecyloxycarbonylethyl)guanine to cytidine at the air/water interface and LB film studied by Fourier transform infrared spectroscopy. AB - Monolayer behavior of a nucleolipid amphiphile, 7-(2 octadecyloxycarbonylethyl)guanine (ODCG), on aqueous cytidine solution was investigated by means of surface-molecular area (pi-A) isotherms. It indicates that molecular recognition by hydrogen bonding is present between ODCG monolayer and the cytidine in subphase. The Fourier transform infrared (FTIR) transmission spectroscopic result indicates that the cytidine molecules in the subphase can be transferred onto solid substrates by Langmuir-Blodgett (LB) technique as a result of the formation of Watson-Crick base-pairing at the air/water interface. Investigation by rotating polarized FTIR transmission also suggests that the headgroup recognition of this amphiphile to the dissolved cytidine influence the orientation of the tailchains. PMID- 12633722 TI - 4f-4f absorption spectra of nine-coordinate Pr (III) and Nd (III) complexes in different environments. AB - Absorption spectroscopic studies on the mixed-ligand complexes of Pr (III) and Nd (III) with 2,2'-bipyridyl and thiocyanate in pyridine, DMSO, DMF and methanol are presented. The oscillator strengths for the hypersensitive and non-hypersensitive transitions have been calculated and variation in the oscillator strength and band shapes with respect to solvent type is rationalized in terms of solvent structures and coordinating properties. A comparison is made with 1,10 phenanthroline complexes and has been shown that bpy is a weaker ligand than phen for these ions. Pyridine has been found most effective in promoting 4f-4f intensity and the increase in the oscillator strength in this solvent is due to dynamic ligand polarization mechanism. PMID- 12633723 TI - 4f-4f hypersensitivity in the absorption spectra and NMR studies on paramagnetic lanthanide chloride complexes with 1,10-phenanthroline in non-aqueous solutions. AB - The optical absorption and NMR studies of trivalent lanthanide chloride complexes with 1,10-phenanthroline (phen) are presented and discussed. The 1H NMR spectra of the complexes of La, Pr, Nd, Eu and Yb have been studied in methanol-d(4). The resonances of phen in the NMR spectra of the paramagnetic complexes have been shifted to lower as well as higher fields, which is a manifestation of dipolar interaction. The H (2) protons of the heterocyclic amine display broad resonances. The degree of broadening in Pr, Nd, and Yb complexes follows the order Pr < Nd < Yb. The inter- and intra-molecular shift ratios show that the paramagnetic shift is predominantly due to dipolar interaction. The electronic spectra of Pr, Nd, Ho and Er complexes have been investigated in methanol, pyridine, DMSO and DMF, which reveal that the hypersensitive transitions exhibit larger variation in oscillator strength values and band shapes. The change in the coordination geometry of the complexes and relative basicity of ligand are found responsible for oscillator strength and band shape variation. The interelectronic repulsion and covalency parameters show covalent nature of bonding between the metal and the ligand. PMID- 12633724 TI - Resolution of multicomponent NMR signals using wavelet compression and immune algorithm. AB - A novel algorithm for fast resolution of multicomponent overlapping nuclear magnetic resonance (NMR) spectrum using the wavelet compression and the immune algorithm (IA) is proposed. Taking advantage of the linear property of the wavelet transform (WT), the overlapping NMR spectrum (antigen) can be compressed by WT before it is input into the immune network, the antibodies are also compressed to the same scheme. After extraction of the compressed information of each component, the resolved NMR spectra can be restored by the WT reconstruction. It was proven that the result is almost the same with the result from an IA, but the calculation speed is much faster. PMID- 12633725 TI - Spectral properties and binding study of DNA complexes with a rigid bisintercalator 1,4-bis((N-methylquinolinium-4-yl)vinyl)benzene. AB - The DNA binding behavior of potentially bisintercalating ligand 1,4-bis((N-methyl quinolinium-4-yl)vinyl)benzene was studied by spectrophotometric titration, circular dichroism and matrix-assisted laser desorption-ionization-time-of-flight (MALDI-TOF) mass spectrometry. The formation of large DNA-ligand aggregates observed at low DNA concentration was ascribed to the interstrand cross-linking due to the bisintercalation and/or electrostatic interactions. On the other hand, a monointercalation was observed at higher DNA concentration and in the presence of higher content of NaCl. Intercalative DNA-ligand complex was featured by red shifted absorption band, modest hypochromicity and the presence of induced CD signal. MALDI-TOF mass spectra of short oligonucleotide-ligand systems revealed the formation of 1:1 complexes of the ligand with duplex and single-stranded oligonucleotides as well as a higher molecular weight species. PMID- 12633726 TI - A new approach to suppress nonlinearity-transparency trade-off through coordination chemistry: syntheses and spectroscopic study on second-order nonlinear optical properties of a series of square-pyramidal zinc(II) complexes. AB - Five new square-pyramidal coordination compounds L x Zn(acac)(2) (1-5) (acac = acetylacetonate; L is a variety of thiosemicarbazones: p dimethylaminobenzaldehyde thiosemicarbazone (1), p-hydroxy-o-methoxybenzaldehyde thiosemicarbazone (2), p-methoxybenzaldehyde thiosemicarbazone (3), p hydroxybenzaldehyde thiosemicarbazone (4), o-hydroxybenzaldehyde thiosemicarbazone (5)) have been synthesized and characterized by 1H NMR, IR, and elemental analysis. All of these compounds exhibit pretty wide transparent ranges in the visible region. Their electronic absorption spectra have been studied experimentally, and theoretically by ZINDO/S calculation. The latter has also been utilized to estimate the extent of intramolecular charge transfer. The MOPAC software package has been used to evaluate their first-order molecular hyperpolarizabilities (beta). All beta values of the five coordination compounds are larger than those of the corresponding thiosemicarbazones. And complex 1 shows the largest beta(0) (39.1 x 10(-30) esu) in the series. PMID- 12633727 TI - A novel vertical attenuated total reflectance photochemical flow-through reaction cell for Fourier transform infrared spectroscopy. AB - A unique photochemical cell design and two experiments are presented, which illustrate the usefulness of flow-through attenuated total reflectance (ATR) Fourier transform infrared (FT-IR) spectroscopy as a technique for investigating photochemical reactions at the mineral-water interface. The kinetics of the photolysis reaction of potassium oxalate (K(2)C(2)O(4)) in a ferric iron solution and oxalate adsorbed onto goethite (alpha-FeOOH) were investigated to show the capabilities of the cell. Due to complicated kinetics, the adsorption experiment demonstrates not only the types of complex problems, that may exist at the mineral-water interface, but also the ability for this novel cell design to address them. PMID- 12633728 TI - Luminescence behavior of a water soluble calix[4]arene derivative complex with terbium ion(III) in gelation solution. AB - The complexation luminescence behavior of a water soluble calix[4]arene derivative, 5,11,17,23-tetra-sulfonate-25,26,27,28-tetra carboxymethoxycalix[4]arene (L) with lanthanoid ion (Tb(3+)) has been investigated in gelation solution at 25 degrees C by using UV-vis and fluorescence spectra. The results obtained indicated that the water soluble calix[4]arene derivative can form an efficient energy transfer complex with terbium ion(III). The fluorescence of L x Tb(3+)complex is partially quenched by gelatin in gelation solution. The quenching intensity is related to the concentration and the hydrolysis degree of gelatin. Absorption and fluorescence spectra analysis show that the -COO(-) groups on gelatin have a definite binding ability to Tb(3+), and then, gelatin could compete binding with calix[4]arene derivative upon complexation with Tb(3+), leading to the relative fluorescence quenching of the formation complex of terbium(III) ion with calix[4]arene derivative. PMID- 12633729 TI - Vibrational spectra, assignments and normal coordinate analysis of 2-amino-5 bromopyridine. AB - The FTIR and laser Raman spectra of 2-amino-5-bromopyridine have been recorded. The observed frequencies were assigned to various modes of vibrations on the basis of normal coordinate calculations, assuming C(s) point group symmetry. The potential energy distribution associated with normal modes is also reported here. The assignment of fundamental vibrations agrees well with the calculated frequencies. PMID- 12633730 TI - Vibrational frequencies and structural determinations of maleonitrile. AB - The vibrational frequencies and corresponding normal mode assignments of maleonitrile are examined theoretically using the GAUSSIAN98 set of quantum chemistry codes. All normal modes were successfully assigned to one of eight types of motion predicted by a group theoretical analysis (C triple bond N stretch, C=C stretch, C-C stretch, C-H stretch, C-H bend, C-C triple bond N bend, C-C triple bond N bend, C-C=C-C torsion) utilizing the C(2v) symmetry of the molecule. The molecular orbitals of maleonitrile are also examined. PMID- 12633731 TI - Homo- and hetero-complexes of exchangeable apolipoproteins in solution and in lipid-bound form. AB - The self-association state of human plasma apolipoprotein E (apoE) in solution and in complexes with dimyristoylphosphatidylcholine (DMPC) varying in stoichiometry was studied in sub-micromolar concentration range by gel filtration, fluorescence anisotropy, fluorescence quenching and energy transfer measurements with apolipoprotein labeled with lysine-specific fluorescent dyes. Together, these results confirm the equilibrium scheme for various apoE structures in solution: oligomer (in aged preparations) <==> 'closed' tetramer <==> 'open' tetramer ('molten globule' state) <==> native or partially denatured monomer <==> fully denatured monomer. Within DMPC:apoE discoidal complex (125:1) the apolipoprotein association state seems to be intermediate between that in solution and in larger vesicular complex (1000:1); for both complexes, the degree of exposure of fluorescein chromophores into water phase decreased. Hetero associates of apoA-I and apoC-III-1 in solution and in the complexes with DMPC appear to behave similarly to apoE. When extrapolated to native HDL particles, 'molten globule' state seems to be a structure responsible for the interaction of exchangeable apolipoproteins with phospholipid. For a first time, the location of various apolipoprotein molecules on disc periphery was confirmed. The lysine residue(s) seems to locate closely to reacting residue(s) within apolipoprotein molecules in associates, however, with different package constraints for discoidal versus vesicular complexes with phospholipid. PMID- 12633732 TI - Antioxidants and fetal protection against ethanol teratogenicity. I. Review of the experimental data and implications to humans. AB - Ethanol is the most common human teratogen, and heavy drinking during pregnancy can result in serious adverse outcomes to the fetus. The cellular mechanisms by which ethanol induces damage in utero are not well understood, while induction of oxidative stress is believed to be one putative mechanism. Our objective is to review the data of antioxidant effects in experimental models of fetal alcohol syndrome. Prior to the description of the available experimental data, we will briefly review the mechanisms leading to ethanol-induced oxidative stress. Ethanol-induced oxidative damage to the fetus could be attenuated by a variety of antioxidants as was documented in whole animal and tissue culture studies. Experiments, retrieved from the literature search, are described and criticized. Although experimental data are still limited, the application of a treatment strategy that includes antioxidants is justified since antioxidant treatment in human pregnancy for pre-eclampsia was demonstrated to be safe and effective. The available experimental evidence and the safety of vitamins C and E in pregnancy suggest that experimental use of antioxidants in alcohol-consuming mothers should be seriously considered to reduce fetal alcohol damage. PMID- 12633733 TI - Cognitive development in preschool children prenatally exposed to PCBs and MeHg. AB - A number of epidemiological studies have shown predictive relationships between prenatal exposure to polychlorinated biphenyls (PCBs) and subtle deficits in cognitive development in infancy through the preschool years [Child Dev. 56 (1985) 853; J. Pediatr. 116 (1990) 38; J. Pediatr. 134 (1999) 33; Toxicol. Lett. 102-103 (1998) 423; Neurotox. 21 (6) (2000) 1029-1038]. However, since not all studies have demonstrated these relationships (J. Pediatr. 119 (1991) 58-63), debate regarding the role of prenatal PCB exposure in cognitive development continues. The current study was designed to provide additional data to assist in resolving this question. Two hundred twelve children enrolled in the Oswego Newborn and Infant Development Project were assessed using the McCarthy Scales of Children's Abilities at 38 months of age, followed by a reassessment at 54 months of age. The relationship between prenatal exposure to PCBs (cord blood PCBs) and McCarthy performance was assessed at both ages after first controlling for a wide range of important predictors of cognitive development, including socioeconomic status (SES), maternal IQ, maternal education, home environment, cigarette smoking, and many others. Cord blood PCBs were statistically significant predictors of small but measurable deficits in McCarthy performance at 38 months of age. Moreover, a significant interaction between cord blood PCBs and maternal hair mercury (MeHg) was found, such that negative associations between prenatal MeHg exposure and McCarthy performance were found in subjects with higher levels of prenatal PCB exposure. No relationship between PCBs and/or MeHg and McCarthy performance was observed when the children were reassessed almost 1.5 years later (54 months of age). Inspection of the age-related trajectory of McCarthy performance revealed that the more highly exposed children caught up with the least exposed children by 54 months. Although the current data partially replicate the findings of Jacobson et al., Patandin et al., and Walkowiak et al. [J. Pediatr. 116 (1990) 38; J. Pediatr. 134 (1999) 33; Lancet 358 (2001) 1602], results reported here suggest that functional recovery may occur. Moreover, the interaction between PCB and MeHg cannot be considered conclusive until it has been replicated in subsequent investigations. PMID- 12633734 TI - Level of prenatal cocaine exposure and infant-caregiver attachment behavior. AB - The objective of this longitudinal prospective cohort study was to determine whether level of prenatal cocaine exposure, or the interaction between level of prenatal cocaine exposure and contextual risk variables, was associated with a higher rate of infant-caregiver insecure attachment and disorganized attachment, or with alterations in infant crying or avoidant behavior, after controlling for prenatal exposure to alcohol, tobacco, and marijuana, the quality of the proximal caregiving environment, and other covariates. Subjects were 154 full-term 12 month-old infants (64 unexposed, 61 with lighter cocaine exposure, 29 with heavier cocaine exposure) and their primary caregivers from low-income, urban backgrounds. Exposure status was determined in the maternity ward by biologic assay (infant meconium and/or maternal or infant urine) and maternal self-report. At the 12-month follow-up visit, infants were videotaped with their primary caregiver in Ainsworth's Strange Situation. Reliable coders masked to exposure status scored videotapes for attachment variables, amount of crying, and level of avoidance. Contrary to popular perceptions, level of prenatal cocaine exposure was not significantly related to secure/insecure attachment status, disorganized attachment status, or rated level of felt security. Foster care status also was not associated with attachment status. However, heavier prenatal cocaine exposure, in interaction with maternal contextual variables (public assistance or multiparity) was associated with alterations in infant socio-affective behavior, including a higher level of behavioral disorganization, more avoidance of the caregiver, and less crying. PMID- 12633735 TI - Solvent ototoxicity in the rat and guinea pig. AB - There is clear evidence that aromatic solvents can disrupt the auditory system in humans and animals. As far as animal models are concerned, solvent-induced hearing loss seems to be species-dependent. Indeed, most published data have been obtained with the rat, which shows mid-frequency cochlear deficits, whereas the guinea pig does not show any permanent hearing loss after solvent exposure. In the current investigation, the effects of two solvents, toluene (600 ppm) and styrene (1000 ppm), were studied in both Long-Evans rats and pigmented guinea pigs exposed 6 h/day for 5 consecutive days. Cochlear function was tested by using distortion product otoacoustic emissions (DPOAE) measured prior to the solvent exposure, 20 min after the end of the exposure and successively at 2 and 4 weeks post-exposure. In addition to cochlear testing, solvent concentrations in blood and urinary metabolites were measured. A cochlear histological analysis was performed at the end of the experiment. No decrease in DPOAE amplitude was observed in the guinea pig, even immediately following the end of exposure. The rat model showed severe disruption of auditory function and cochlear pathology, whereas the guinea pig had no disruption of DPOAE or cochlear pathological alterations. Therefore, the vulnerability of the cochlear function was strictly dependent on the species. As expected, an important difference in the styrene concentration in blood was observed: the solvent concentrations were fourfold higher in the rat than in the guinea pig. Therefore, it is clear that a pharmacokinetic or an uptake difference might explain the difference in susceptibility observed between the two species. Moreover, the metabolism pathways of the solvents were different depending on the species. Attempts to explain differences of vulnerability between the rat and guinea pig are addressed in the present paper. PMID- 12633736 TI - Chlorpyrifos exposure of developing zebrafish: effects on survival and long-term effects on response latency and spatial discrimination. AB - Chlorpyrifos (CPF) is a widely used insecticide, which has been shown to interfere with neurobehavioral development. Rat models have been key in demonstrating that prenatal CPF exposure causes choice accuracy deficits and motor alterations, which persist into adulthood. Complementary nonmammalian models can be useful in determining the molecular mechanisms underlying the persisting behavioral effects of developmental CPF exposure. Zebrafish with their clear chorion and extensive developmental information base provide an excellent model for assessment of molecular processes of toxicant impacted neurodevelopment. To facilitate the use of the zebrafish model and to compare it to the more typical rodent models, the behavioral phenotype of CPF toxicity in zebrafish must be well characterized. Our laboratory has developed methods for assessing spatial discrimination learning in zebrafish, which can differentiate response latency from choice accuracy in a three chambered fish tank. Low and high doses of CPF (10 and 100 ng/ml on days 1-5 postfertilization) both had significant persisting effects on both spatial discrimination and response latency over 18 weeks of testing. The high, but not the low dose, significantly accelerated mortality rates of the fish during the study from 20-38 weeks of age. Developmental exposure to either 10 or 100 ng/ml of CPF caused significant spatial discrimination impairments in zebrafish when they were adults. The impairment caused by 10 ng/ml was seen during early but not later testing, while the impairment caused by 100 ng/ml became more pronounced with continued testing. The higher dose caused a more pervasive impairment. The 10 and 100 ng/ml doses had opposite effects on response latency. The low 10 ng/ml dose significantly slowed response latency, while the high 100 ng/ml dose significant increased response latency. Both of these effects diminished with continued testing. CPF exposure during early development caused clear behavioral impairments, which lasted throughout adulthood in zebrafish. The molecular mechanisms by which early developmental CPF exposure produces these behavioral impairments expressed in adulthood can now be studied in the zebrafish model. PMID- 12633737 TI - Effects of chronic prenatal ethanol exposure on cGMP content and glutamate release in the hippocampus of the neonatal guinea pig. AB - The glutamate-N-methyl-D-aspartate (NMDA) receptor-nitric oxide synthase (NOS) cGMP signal transduction system plays key neurotrophic and intercellular communication roles in the hippocampus. In the guinea pig, chronic prenatal ethanol exposure (CPEE), via maternal ethanol administration, suppresses the hippocampal glutamate-NMDA receptor-NOS pathway in the near-term fetus and decreases stimulated glutamate release in the hippocampus of young postnatal offspring, with no effect on NMDA receptor number or NOS activity. At present, the effect of CPEE on cGMP, a key second messenger of the glutamate signal transduction system, in the hippocampus is not known. The objective of this study was to test the hypothesis that CPEE suppresses the hippocampal glutamate signal transduction system in the neonatal guinea pig at the levels of cGMP content and glutamate release. Timed pregnant guinea pigs received chronic oral administration of 4 g ethanol/kg maternal body weight/day, isocaloric sucrose/pair-feeding, or water treatment throughout gestation. CPEE decreased brain and hippocampal weights at postnatal day (PD) 1 and PD 5 (P<.05). CPEE did not affect basal, NMDA (1, 10, or 100 microM)-stimulated, or K(+) (15 or 30 mM) stimulated cGMP content in transverse hippocampal slices at PD 1 or 5. At 60 mM K(+), however, CPEE decreased stimulated hippocampal cGMP content at PD 1 (P<.05) and increased stimulated cGMP content at PD 5 (P<.05). In transverse hippocampal slices, CPEE did not affect basal or K(+) (40 or 45 mM)-stimulated glutamate release at PD 1 or 5, or NMDA (50 microM)-stimulated glutamate release at PD 1, but did decrease NMDA (50 microM)-stimulated glutamate release at PD 5 (P<.05). The data demonstrate that the effects of CPEE on stimulated cGMP content and glutamate release in the hippocampus of the neonatal guinea pig are stimulating agent- and age-dependent. PMID- 12633738 TI - Neurotoxic effects of mercury on auditory cortex networks growing on microelectrode arrays: a preliminary analysis. AB - Mercury is known to cause sensorineural hearing loss and impaired speech perception. However, there is still a lack of a quantitative description of mercury toxicity on central auditory structures. This is a preliminary study using the novel technique of microelectrode array (MEA) recordings to evaluate acute and chronic neurotoxic effects of mercury on auditory cortex networks (ACNs) in vitro. Morphological and electrophysiological effects of mercuric chloride (HgCl(2)) were studied. Neurons dissociated from auditory cortices of 14 day-old mouse embryos were grown on photoetched MEAs containing 64 transparent indium-tin oxide (ITO) electrodes. For acute electrophysiological experiments, the spontaneous spiking and bursting activity from ACNs were compared before and after application of HgCl(2). For chronic electrophysiological experiments, auditory cortex cultures were treated with various concentrations of HgCl(2) from the day of seeding, and were tested 4 weeks later for the presence of spontaneous activity. Morphological analysis was conducted on 8-day-old ACNs treated with HgCl(2) for 3 days. Results of acute experiments indicated that <75 mM of HgCl(2) had an excitatory effect of variable magnitude on the spontaneous activity of ACNs; however, concentrations above 100 microM completely and irreversibly inhibited spike and burst activity. Chronic exposure of ACNs to 10 microM HgCl(2) completely blocked the spontaneous activity. Morphological analysis indicated that 10 microM HgCl(2) caused neuronal cell death in 3 days. It is concluded that HgCl(2) has a more toxic effect on auditory networks when exposed chronically, and the levels of mercury showing toxic effects on ACNs are within the dose range shown to cause neurologic symptoms in humans. PMID- 12633740 TI - Impairment of the EGF signaling pathway by the oxidative stress generated with UVA. AB - Ultraviolet A (UVA) is a component of sunlight reaching the surface of the earth and involved in photodegenerescence and photocarcinogenesis. The effect of UVA was investigated on the EGF-induced activation of the signaling kinase ERK and the transcription factors AP1, NFkappaB, and STAT1. UVA prevented the Epidermal Growth Factor (EGF)-induced stimulation of ERK in a dose-dependent manner within the range of 1.5-9 J/cm(2). Concomitantly, the DNA binding activity of AP1, NFkappaB, and STAT1 under EGF were markedly inhibited by UVA within the same dose range. UVA by itself induced an activation of ERK activity, and a stimulation of AP1, NFkappaB, and STAT1 binding activity. UVA decreased EGF binding in a dose dependent manner. Furthermore, the highest dose of UVA (9 J/cm(2)) prevented the EGF-induced Tyr-phosphorylation of the EGF-receptor (EGF-R). The generation of reactive oxygen species (ROS), as assessed by the fluorescent probe dichloro fluorescein, showed an additive effect of EGF and UVA, within the studied range of UVA doses. Finally, the antioxidant Vitamin E prevented the inhibitory effect of UVA on ERK, AP1, NFkappaB, and STAT1. These results demonstrate that an overproduction of ROS, initiated by two different and successive triggering agents such as UVA and EGF, leads to inactivation of the EGF signaling pathway. This inhibition of gene expression control by EGF might play a role in the photodegenerative processes observed after exposition of skin cells to solar radiation. PMID- 12633741 TI - Photo-oxidation of cells generates long-lived intracellular protein peroxides. AB - Singlet oxygen is generated by several cellular, enzymatic, and chemical reactions as well as by exposure to UV or visible light in the presence of a sensitizer. Consequently, this oxidant has been proposed to be a damaging agent many pathologies. Proteins are major targets for singlet oxygen as a result of their abundance and high rate constants for reaction. In this study, we show that illumination of viable rose bengal-loaded THP-1 (human monocyte-like) cells with visible light gives rise to intracellular protein-derived peroxides. The peroxide yield increases with illumination time, requires the presence of rose bengal, is enhanced in D(2)O, and is decreased by azide, consistent with the mediation of singlet oxygen. The concentration of peroxides detected, which is not affected by glucose or ascorbate loading of the cells, corresponds to about 1.5 nmoles peroxide per 10(6) cells, or 10 nmoles/mg cell protein, and account for up to approximately 15% of the O(2) consumed by the cells. Similar peroxides have been detected on isolated cellular proteins exposed to light in the presence of rose bengal and oxygen. After cessation of illumination, cellular protein peroxide levels decrease with t(1/2) about 4 h at 37 degrees C. Decomposition of protein peroxides formed within cells, or on isolated cellular proteins, by metal ions gives rise to radicals as detected by EPR spin trapping. These studies demonstrate that exposure of intact cells to visible light in the presence of a sensitizer leads to novel long-lived, but reactive, intracellular protein peroxides via singlet oxygen-mediated reactions. PMID- 12633742 TI - The relationship between the antioxidant and the antibacterial properties of galloylated catechins and the structure of phospholipid model membranes. AB - The effects of four catechins, (+)-catechin (C), (-)-epicatechin (EC), (-) epicatechin gallate (ECG), and (-)-epigallocatechin gallate (EGCG), on the physical properties of phospholipid model membranes and the correlation to their antioxidant and antibacterial capacities have been studied by using differential scanning calorimetry (DSC), fluorescence spectroscopy, infrared spectroscopy (IR), AAPH-induced oxidation, and leakage experiments. DSC data revealed that galloylated catechins, especially ECG, affected the physical properties of both the phosphatidylcholine (PC) and phosphatidylethanolamine (PE) bilayers dramatically. Galloylated catechins showed higher phospholipid/water partition coefficients than their homologues and were immersed in the phospholipid palisade intercalating within the hydrocarbon chains, ECG being at the deepest position. In contrast, nongalloylated catechins presented a shallow location close to the phospholipid/water interface. ECG also exhibited the highest antioxidant capacity against lipid peroxidation, which correlated with its strong effect on DPH fluorescence anisotropy (as observed by the increase of the lipid order of fluid PC bilayers) and with the presence of highly cooperative transitions as seen by DSC. We propose that the high antioxidant capacity of some galloylated catechins such as ECG could be partially due to the formation of membrane structures showing resistance to detergent solubilization and in which the phospholipids have tightly packed acyl chains and highly hydrated phosphate groups. Significantly, PE was found to be essential to the promotion of carboxyfluorescein leakage from bacterial model membranes by galloylated catechins, indicating that their bactericidal activity, at least at the membrane level, could be due to the specific effect of these catechins on PE. PMID- 12633743 TI - Vitamin E deficiency reduces surfactant lipid biosynthesis in alveolar type II cells. AB - Reactive oxygen species play an important role in development of lung injury. Neonates exhibit a high risk of developing acute and/or chronic lung disorder, often associated with surfactant deficiency, and in parallel they show low vitamin E concentration. We investigated whether the vitamin E status of adult rats affects the content of phospholipids (PL) in bronchoalveolar lavage and alveolar type II cells. Phosphatidylcholine (PtdCho) is the dominant and functional most important PL in lung surfactant. Therefore, we determined its formation via de novo synthesis and reacylation of lyso-PtdCho in type II cells. Vitamin E depletion caused a decrease of PL content in bronchoalveolar lavage and type II cells and decreased glycerol-3-phosphate O-acyltransferase (G3P-AT) activity, de novo synthesis of PtdCho, and reacylation of lyso-PtdCho in type II cells. Preincubation of type II cell homogenates with dithiothreitol restored the activity of G3P-AT and de novo synthesis but inhibited reacylation. Reacylation was strongly reduced by chelerythrine-mediated inhibition of protein kinase C. We conclude that antioxidant and PKC-modulating properties of vitamin E regulate de novo synthesis of PtdCho and reacylation of lyso-PtdCho in alveolar type II cells. Vitamin E depletion reduced the two pathways of PL synthesis and caused a decrease of PL content in alveolar surfactant of rats. PMID- 12633744 TI - Intracellular metal ion chelators inhibit TNFalpha-induced SP-1 activation and adhesion molecule expression in human aortic endothelial cells. AB - Endothelial adhesion molecule expression and monocyte recruitment are causal events in human atherosclerosis, and are believed to be caused, in part, by oxidative stress. Because redox-active transition metal ions, such as iron and copper, play an essential role in the generation of free radicals and the initiation and propagation of lipid peroxidation, we hypothesized that transition metal ions may also be involved in endothelial activation. Therefore, we investigated the effects of the intracellular iron-chelator, desferrioxamine (DFO), and the intracellular copper-chelator, neocuproine (NC), on TNFalpha induced expression of adhesion molecules in human aortic endothelial cells (HAEC). Treatment of HAEC with DFO (0.01-0.1 mM) or NC (0.1 and 0.5 mM) time- and dose-dependently inhibited TNFalpha-induced protein expression of E-selectin, vascular cell adhesion molecule-1 (VCAM-1), and intercellular adhesion molecule-1 (ICAM-1). In contrast, iron-saturated DFO and the extracellular copper chelator, bathocuproinedisulfonic acid, had no effect on adhesion molecule expression. DFO and NC also dose-dependently inhibited TNFalpha-induced upregulation of adhesion molecule mRNA levels. Furthermore, treatment of HAEC with 0.5 mM DFO or NC completely inhibited TNFalpha-induced activation of the transcription factor, specificity protein-1 (SP-1), but only partially inhibited or did not affect activation of other transcription factors known to regulate adhesion molecule expression, i.e., nuclear factor kappaB (NFkappaB), activator protein-1 (AP-1), and interferon regulatory factor-1 (IRF-1). Finally, inhibition of endothelial nitric oxide synthase with N-nitro-L-arginine methylester (0.5 mM) did not attenuate the inhibitory effects of the metal ion chelators on adhesion molecule expression. Our data suggest that intracellular, but not extracellular, transition metal ions mediate inflammatory cytokine-induced SP-1 activation and adhesion molecule expression in endothelial cells. PMID- 12633745 TI - Sequential inactivation of reactive oxygen species by combined overexpression of SOD isoforms and catalase in insulin-producing cells. AB - Insulin-producing cells show very low activity levels of the cytoprotective enzymes catalase, glutathione peroxidase, and superoxide dismutase. This weak antioxidative defense status has been considered a major feature of the poor resistance against oxidative stress. Therefore, we analyzed the protective effect of a combined overexpression of Cu,ZnSOD or MnSOD together with different levels of catalase. Catalase alone was able to increase the resistance of transfected RINm5F insulin-producing tissue culture cells against H(2)O(2) and HX/XO, but no protection was seen in the case of menadione. In combination with an increase of the MnSOD or Cu,ZnSOD expression, the protective action of catalase overexpression could be further increased and extended to the toxicity of menadione. Thus, optimal protection of insulin-producing cells against oxidative stress-mediated toxicity requires a combined overexpression of both superoxide- and hydrogen peroxide-inactivating enzymes. This treatment can compensate for the constitutively low level of antioxidant enzyme expression in insulin-producing cells and may provide an improved protection in situations of free radical mediated destruction of pancreatic beta cells in the process of autoimmune diabetes development. PMID- 12633746 TI - Antioxidant role of N-acetyl cysteine isomers following high dose irradiation. AB - High dose, acute radiation exposure, as in radiation accidents, induces three clinical syndromes that reflect consequences of oxidative protein, lipid, and DNA damage to tissues such as intestine, lung, and liver. In the present study, we irradiated C57BL/6 mice with 18 Gy whole-body radiation (XRT) and evaluated N acetyl cysteine (NAC) isomers LNAC and DNAC as potential radioprotectors under conditions that would model the gastrointestinal syndrome. We focused on tissues thought not immediately involved in the gastrointestinal syndrome. Both LNAC and DNAC protected the lung and red blood cells (RBC) from glutathione (GSH) depletion following radiation exposure. However, only LNAC also supplemented the spleen GSH levels following XRT. Protection from increased malondialdehyde (MDA) levels (lung) and increased 8-hydroxy-deoxyguanosine (8-oxo-dG) presence (liver) following XRT was observed with treatment by either isomer of NAC. These results imply that either NAC isomer can act as a radioprotectant against many aspects of oxidative damage; chirality is only important for certain aspects. This pattern would be consistent with direct action of NAC in many radioprotection and repair processes, with a delimited role for NAC in GSH synthesis in some aspects of the problem. PMID- 12633747 TI - NF-kappaB activation is detrimental in arginine-induced acute pancreatitis. AB - The transcription factor nuclear factor kappaB (NF-kappaB) has been shown to have a critical role in the pathogenesis of sodium taurocholate- and cerulein-induced acute pancreatitis by regulating the expression of many proinflammatory genes in the pancreas. Heat shock proteins (HSPs), on the other hand, protect the pancreas against cellular damage. The aims of the present study were: (i) to investigate pancreatic NF-kappaB activation, proinflammatory cytokine synthesis, and cytoprotective HSP induction during L-arginine- (Arg-) induced acute pancreatitis in rats, and (ii) to establish whether pretreatment with pyrrolidine dithiocarbamate (PDTC) or methylprednisolone (MP) can block the activation of pancreatic NF-kappaB and determine their effects on the severity of Arg-induced acute pancreatitis. The dose-response (3 or 4 g/kg) and time-effect (0.5-96 h) curves relating to the action of Arg on pancreatic NF-kappaB activation and IL 1beta, TNF-alpha, HSP60, and HSP72 synthesis were evaluated. Various doses of PDTC or MP were administered 1 h before the induction of pancreatitis. We demonstrated that Arg specifically and dose-dependently induces pancreatitis, activates NF-kappaB (only the 3 g/kg dose) and proinflammatory cytokine synthesis, and increases the expressions of HSP60 and HSP72 in the pancreas of rats. The lower dose of Arg induced a less severe pancreatitis, but larger increases in the levels of HSPs. The present work supports and extends earlier observations that NF-kappaB activation is a common mechanism in acute pancreatitis, although it is dose dependent and occurs at a later stage in Arg induced pancreatitis as compared with other models. PDTC and MP pretreatment dose dependently blocked NF-kappaB activation and proinflammatory cytokine expression and ameliorated many of the examined laboratory (the pancreatic weight/body weight ratio, the pancreatic myeloperoxidase activity, the pancreatic contents of protein, amylase and trypsinogen, the degrees of lipid peroxidation and protein oxidation, and the nonprotein sulfhydryl group content) and morphological parameters of the disease. These findings suggest that pretreatment with PDTC or MP has an anti-inflammatory effect during Arg-induced pancreatitis, which is at least partly mediated by the inhibition of NF-kappaB activation and proinflammatory cytokine synthesis. The increased levels of HSPs most probably act to limit the severity of the disease. PMID- 12633748 TI - Correlation between sonochemistry of surfactant solutions and human leukemia cell killing by ultrasound and porphyrins. AB - The synergistic effect of ultrasound and drugs on cells is known as sonodynamic therapy. The use of sonodynamic therapy for the potential clinical treatment of certain tumors is promising, however, the mechanism of sonodynamic therapy could be due to either sonomechanical and/or sonochemical effects on the cells. The aim of the current study is to determine the importance of the sonochemical mechanism for sonodynamic therapy. Sonochemical effects arise from the formation of radical species following collapse of cavitation bubbles. The synergistic effect of ultrasound (47 kHz) and analogues of a gallium-porphyrin derivative (ATX-70) on cytolysis of Human leukemia cells (HL-525 and HL-60) suspended in a cell culture medium were studied. Organic surfactants preferentially accumulate and subsequently decompose at the gas/solution interface of cavitation bubbles, producing secondary radicals that can diffuse to the bulk solution. The gallium porphyrin analogues used in the current study possess two n-alkyl side chains (ATX-C(x), where x = number of carbon atoms, ranging from x = 2 to x = 12). By varying the n-alkyl chain length, thereby modifying the surfactant properties of the ATX-C(x) derivatives, cell killing in relation to the accumulation of ATX C(x) derivatives at the gas/solution interface of cavitation bubbles was determined. Following sonolysis in the presence of ATX-C(x), a strong correlation for the yield of carbon-centered radicals and cell killing was observed. These results support the hypothesis that a sonochemical mechanism is responsible for the synergistic effect of ultrasound and ATX-C(x) on HL-525 and HL-60 cells. PMID- 12633749 TI - Influence of epithelial lining fluid lipids on NO(2)-induced membrane oxidation and nitration. AB - Within the pulmonary epithelial lining layer (ELF), antioxidants such as ascorbic acid (AH(2)) and glutathione (GSH) react with inhaled nitrogen dioxide ((*)NO(2)) to produce reactive oxygen species (ROS) that induce cellular oxidation. Because the ELF contains unsaturated fatty acids (UFA), which potentially react with (*)NO(2) and/or the antioxidant-derived ROS, we studied the influence of aqueous phase model UFA [egg phosphatidylcholine (EggPC) liposomes] on exposure-induced oxidation and nitration of membranes. Our lung surface model used gas phase (*)NO(2) exposures of immobilized red cell membranes (RCM) overlaid with defined aqueous phases. Acetyl cholinesterase (AChE) activity, TBARS, and 3-nitrotyrosine (3-NT) were used to assess protein and lipid oxidation and RCM nitration, respectively. During (*)NO(2) exposure, AH(2) and GSH induced AChE loss and TBARS, which were unchanged with buffer only. Exposures of EggPC generated extensive TBARS but not AChE loss; addition of AH(2)/GSH to EggPC resulted in smaller AChE declines and fewer TBARS. 3-NT formation occurred with or without EggPC, low concentration antioxidants, SOD, catalase, or DTPA, but was inhibitable by desferrioxamine or high antioxidant concentrations. The data suggest that reaction/diffusion limitations govern (*)NO(2) distribution, that (*)NO(2) per se directly nitrates tyrosine residues within hydrophobic regions, and that the induction of secondary oxidative processes is dependent on nonlinear relationships among (*)NO(2) flux rates, antioxidant concentrations, and diffusivity of secondary reactive species. PMID- 12633750 TI - Fluorobenzo[a]pyrenes as probes of the mechanism of cytochrome P450-catalyzed oxygen transfer in aromatic oxygenations. AB - Fluoro substitution of benzo[a]pyrene (BP) has been very useful in determining the mechanism of cytochrome P450-catalyzed oxygen transfer in the formation of 6 hydroxyBP (6-OHBP) and its resulting BP 1,6-, 3,6-, and 6,12-diones. We report here the metabolism of 1-FBP and 3-FBP, and PM3 calculations of charge densities and bond orders in the neutral molecules and radical cations of BP, 1-FBP, 3-FBP, and 6-FBP, to determine the mechanism of oxygen transfer for the formation of BP metabolites. 1-FBP and 3-FBP were metabolized by rat liver microsomes. The products were analyzed by HPLC and identified by NMR. Formation of BP 1,6-dione and BP 3,6-dione from 1-FBP and 3-FBP, respectively, can only occur by removal of the fluoro ion from C-1 and C-3, respectively, via one-electron oxidation of the substrate. The combined metabolic and theoretical studies reveal the mechanism of oxygen transfer in the P450-catalyzed formation of BP metabolites. Initial abstraction of a pi electron from BP by the [Fe(4+)=O](+)(*) of cytochrome P450 affords BP(+)(*). This is followed by oxygen transfer to the most electropositive carbon atoms, C-6, C-1, and C-3, with formation of 6-OHBP (and its quinones), 1 OHBP, and 3-OHBP, respectively, or the most electropositive 4,5-, 7,8-, and 9,10- double bonds, with formation of BP 4,5-, 7,8-, or 9,10-oxide. PMID- 12633751 TI - Correlation of iron and zinc levels with lesion depth in newly formed atherosclerotic lesions. AB - Several studies have indicated a relationship between body iron content and cardiovascular disease, although other studies have not. There are also suggestions that zinc has an antioxidant and antiatherosclerotic effect. We have used Nuclear Microscopy, using the combination of Scanning Transmission Ion Microscopy (STIM), Rutherford Backscattering Spectrometry (RBS), and Proton Induced X-ray Emission (PIXE) to map and quantify iron and zinc levels in newly formed atherosclerotic lesions. Sixteen New Zealand White rabbits fed on a high cholesterol diet were divided into four groups of 4 rabbits each. Six weeks into the high cholesterol diet, two groups were treated with the iron chelating agent desferrioxamine, for 2 weeks and 4 weeks, respectively, by surgically implanting with Alzet osmotic pumps (Alza Corporation, Palo Alto, CA, USA) containing desferal (0.5 g/ml). The other two groups served as controls, and were surgically implanted with osmotic pumps containing saline. Tissue sections were taken from the aortic arch, flash frozen, and air-dried. Analysis of atherosclerotic lesions indicated a trend (p =.07) to a reduction in the progression of the lesion after 4 weeks of desferrioxamine treatment. For each of the control and desferrioxamine treated animals however, the more extensive lesions contained a higher concentration of iron and a lower concentration of zinc. Our results are consistent with the view that early lesion formation may be accelerated by free radical production caused by increased iron levels, that zinc might antagonize such effects, and that more prolonged desferal treatment might have an antiatherosclerotic effect. PMID- 12633753 TI - EPR studies of in vivo radical production by lipopolysaccharide: potential role of iron mobilized from iron-nitrosyl complexes. AB - Although oxidative stress has been implicated in the pathogenesis of sepsis, there is little evidence for the formation of radicals other than nitric oxide in its experimental models. Here we used low temperature EPR and EPR spin trapping to monitor nitric oxide and secondary radical formation in blood, liver, and bile samples from rats treated with a low lipopolysaccharide (LPS) dose (0.25 mg) and with dimethyl sulfoxide (DMSO) and the spin trap alpha-(4-pyridyl 1-oxide)- N-t butylnitrone (POBN). The results showed that production of secondary radicals triggered by LPS is delayed in regard to maximum nitric oxide synthesis and is iron-dependent. One of the secondary produced radicals was identified as the hydroxyl radical. Its formation is proposed to occur because of the mobilization of redox-active iron required to repair the nitrosyl complexes produced by LPS. The results suggest that iron chelation may be a useful adjuvant therapy for treating sepsis. PMID- 12633752 TI - Attenuated mild colonic inflammation and improved survival from severe DSS colitis of transgenic Cu/Zn-SOD mice. AB - Mucosal tissue damage in chronic inflammatory bowel disease (IBD) is partly caused by an enduring exposure to excessive amounts of reactive oxygen metabolites (ROM). To protect themselves from the toxic effects of ROM, most intestinal cell types constitutively express the highly specific, key ROM neutralizing cytosolic enzyme Cu/Zn-superoxide dismutase (SOD). Under inflammatory conditions, however, its protein and activity levels have consistently been reported as being decreased. To elucidate a direct functional relationship between intracellular Cu/Zn-SOD expression and intestinal inflammation, we investigated the effects of transgenic human Cu/Zn-SOD overexpression in acute and chronic murine dextran sodium sulfate (DSS)-induced colitis. When subjected to a mild form of acute colitis, the Cu/Zn-SOD overexpressing mice showed a significantly lower colonic activity of neutrophilic myeloperoxidase (MPO) than their nontransgenic littermates. This difference was particularly evident in the male animals. In contrast, a severe acute colitis did not lead to any differences in MPO activity between both groups. Yet, when the animals were subsequently allowed to recover, MPO levels were again significantly lower in the transgenes, suggesting an involvement of Cu/Zn-SOD in, particularly, the clearance of neutrophils. Specific, immunohistochemical identification of neutrophils confirmed the validity of the MPO activity measurements. In addition, transgenic animals showed a remarkable survival benefit from severe DSS colitis over their nontransgenic littermates, particularly during or shortly after the acute inflammatory phase. During the chronic inflammatory phase, which was not characterized by massive neutrophil infiltration, no effects of Cu/Zn-SOD overexpression were noted. Paradoxically, overexpression of Cu/Zn-SOD did not obviously improve the colitis-related (oxidative) injury or symptoms at any stage of the experiment. Surprisingly, however, we did observe a pronounced male gender preference for DSS susceptibility that was reflected by increased male colitis mortality. Our findings provide direct in vivo evidence for a protective, neutrophil-related role for Cu/Zn-SOD in intestinal inflammation. As such, they support the concept of SOD-based (adjunct) antioxidant treatment strategies for inflammatory bowel disease. PMID- 12633754 TI - Paraoxonase (PON1) deficiency is associated with increased macrophage oxidative stress: studies in PON1-knockout mice. AB - Human serum paraoxonase (PON1), an HDL-associated esterase, protects lipoproteins against oxidation, probably by hydrolyzing specific lipid peroxides. As arterial macrophages play a key role in oxidative stress in early atherogenesis, the aim of the present study was to examine the effect of PON1 on macrophage oxidative stress. For this purpose we used mouse arterial and peritoneal macrophages (MPM) that were harvested from two populations of PON1 knockout (KO) mice: one on the genetic background of C57BL/6J (PON1(0)) and the other one on the genetic background of apolipoproteinE KO (PON1(0)/E(0)). Serum and LDL, but not HDL, lipids peroxidation was increased in PON1(0), compared to C57BL/6J mice, by 84% and by 220%, respectively. Increased oxidative stress was shown in peritoneal and in arterial macrophages derived from either PON1(0) or PON1(0)/E(0) mice, compared to their appropriate controls. Macrophage oxidative stress was expressed by increased lipid peroxides content in MPM from PON1(0) and from PON1(0)/E(0) mice by 48% and by 80%, respectively, and by decreased reduced glutathione (GSH) content, compared to the appropriate controls. Furthermore, increased capacity of MPM from PON1(0) and PON1(0)/E(0) mice to oxidize LDL (by 40% and by 19%, respectively) and to release superoxide anions was observed. In accordance with these results, PON1(0) mice MPM exhibited 130% increased translocation of the cytosolic p47phox component of NADPH-oxidase to the macrophage plasma membrane, suggesting increased activation of macrophage NADPH-oxidase in PON1(0) mice, compared to control mice MPM. The increase in oxidative stress in PON1-deficient mice was observed despite the presence of the two other members of the PON gene family. PON2 and PON3 activities and mRNA expression were both found to be present in PON1-deficient mice MPM. Upon incubation of PON1(0)/E(0) derived macrophages with human PON1 (7.5 arylesterase units/ml), cellular peroxides content was decreased by 18%, macrophage superoxide anion release was decreased by 33%, and macrophage-mediated oxidation of LDL was reduced by 22%. Finally, a 42% increase in the atherosclerotic lesion area was observed in PON1(0)/E(0) mice, in comparison to E(0) mice under regular chow diet. We thus concluded that PON1 can directly reduce oxidative stress in macrophages and in serum, and that PON1-deficiency results in increased oxidative stress not only in serum, but also in macrophages, a phenomenon that can contribute to the accelerated atherosclerosis shown in PON1-deficient mice. PMID- 12633756 TI - Breast cancer as a model of realistic challenges in pharmacogenomics. PMID- 12633755 TI - Fluorescence imaging of lipid peroxidation in isolated rat lungs during nonhypoxic lung ischemia. AB - We have shown previously that ischemia results in reactive oxygen species production by lung endothelium that occurs within 3-5 s after flow cessation and is followed by lipid peroxidation at 15-30 min as determined by assay of thiobarbituric acid-reactive substances, conjugated dienes, and protein carbonyls in lung homogenate. The present study evaluated membrane lipid peroxidation in isolated, ventilated rat lungs using a fluorescence imaging method that permits continuous observation of pulmonary subpleural microvascular endothelial cells in situ. Diphenyl-1-pyrenylphosphine (DPPP), a fluorescent probe which localizes in the plasma membrane and shows increased fluorescence emission after its oxidation by lipid hydroperoxides, was used for detection of membrane lipid peroxidation. Compared to continuously perfused control lungs, endothelial cell DPPP fluorescence increased significantly within 1 min of ischemia (i.e., flow cessation); these changes were prevented by pretreatment with 0.5 mM alpha tocopherol succinate (vitamin E) added to the perfusate. Increased DPPP fluorescence was confirmed by spectrofluorometry of lipid extracts of lung homogenates. These data indicate that DPPP can be used for the real-time detection of lipid peroxidation in an intact organ. Ischemia results in peroxidation of the pulmonary microvascular endothelial cell membrane and this insult can be detected as early as 1 min after the onset of ischemia compatible with a radical-mediated process. PMID- 12633757 TI - Dyslipidemia in pediatric nephrotic syndrome: causes revisited. PMID- 12633758 TI - Comparison of lactate, bilirubin and hemoglobin F concentrations obtained by the ABL 700 series blood gas analyzers with laboratory methods. AB - OBJECTIVE: To compare lactate, bilirubin and Hemoglobin F concentrations obtained on ABL 700 series blood gas analyzers with those from laboratory methods. DESIGN AND METHOD: Pooled neonatal plasma, cord blood and adult plasma samples were used for comparison of bilirubin, hemoglobin F and lactate concentrations respectively. RESULTS: Results obtained on the ABL 700 series compared favorably (Deming regression slopes 0.97-1.13) with those from laboratory methods. For lactate ABL (y) = 1.13 Vitros (x) -0.43 with a CI (slope) of 1.10 to 1.16, CI (int) of -0.61 to -0.28. For hemoglobin F ABL(y) = 1.11 Variant (x) -8.0 with a CI (slope) of 0.88 to 1.33, CI (int) of -25.3 to 9.3. The three bilirubin comparisons are as follows: 1) Unistat (y) = 1.10 Vitros (x) -16.12 with CI (slope) of 1.06 to 1.14 and CI (int) of -25.3 to -6.9. 2), ABL (y) = 0.97 Vitros(x) -10.16 with CI (slope) of 0.94 to 1.00 and CI (int) of -17.6 to 2.73) Unistat (y) = 1.14 (x)-4.58 with CI (slope) of 1.09 to 1.18 and CI (int) of -13.6 to 4.5. CONCLUSION: The ABL 700 series gave comparable results for lactate, bilirubin and hemoglobin F with laboratory methods and may be used in patient care. PMID- 12633759 TI - Hormone stability in human whole blood. AB - OBJECTIVE: To determine whether significant changes in the plasma concentrations of 17 hormones occur when human whole blood is held at 4 or 24 degrees C for up to 24 h before separation of the plasma fraction. DESIGN AND METHODS: Blood samples (EDTA) from healthy human volunteers were held at 4 degrees C or 24 degrees C for 0.5, 6 or 24 h before separation. Plasma concentrations of ACTH, aldosterone, gonadotrophin alpha-subunits, AVP, C-peptide, estradiol, FSH, GH, glucagon, IGF-1, IGFBP3, insulin, leptin, LH, prolactin, PTH and VIP were measured and the results compared to baseline values. Nonlinear regression was used to test for a significant mean rate of change. The time interval for median concentrations to change by 10% was determined. RESULTS: Significant changes were observed for ACTH (decrease at 18.6 hr, 4 degrees C; 17.5 hr, 24 degrees C); AVP (increase at 2.6 h, 24 degrees C); insulin (decrease at 16.8 hr, 4 degrees C; 16.9 hr, 24 degrees C) and VIP (increase at 18.6 h, 24 degrees C). No changes were detected for the remaining analytes.B CONCLUSIONS: The measurement of some hormones is compromised by a delay in plasma separation from normal human blood. While many hormones appear stable in normal whole blood, we recommend that processing occurs without delay. PMID- 12633760 TI - The susceptibility of lipoprotein(a) to copper oxidation is correlated with the susceptibility of autologous low density lipoprotein to oxidation. AB - OBJECTIVES: Lipoprotein(a) [Lp(a)] can be oxidized by copper in vitro in a way comparable to low-density lipoprotein (LDL). We sought to determine whether the susceptibility of Lp(a) to oxidation is correlated with the susceptibility of autologous heterogeneous LDL, with apolipoprotein(a) [apo(a)] molecular size, or with both factors. DESIGN AND METHODS: We examined shifts in electrophoretic mobility of Lp(a) and LDL caused by copper oxidation in plasma samples from 81 healthy men. The effect of copper oxidation on different-sized apo(a) was also evaluated. RESULTS: There was a close correlation between the relative electrophoretic mobilities of oxidized Lp(a) and oxidized LDL in subjects, especially with small-sized apo(a) (n = 25, r = 0.72, p < 0.0001). Oxidative processes in Lp(a) resulted in the degradation of large-, but not small-sized apo(a). CONCLUSIONS: The susceptibility of Lp(a) to oxidation is correlated with that of autologous LDL. Large-sized apo(a) may be involved in the Lp(a) oxidation. PMID- 12633761 TI - One-step quantitative thyrotropin assay for the detection of hypothyroidism in point-of-care conditions. AB - OBJECTIVES: Different screening strategies for early diagnosis of hypothyroidism have been discussed increasingly. We demonstrate the applicability of a miniaturized microparticle assay format for rapid and quantitative determination of increased thyrotropin (TSH) concentrations in serum. DESIGN AND METHODS: Porous microparticles were used as solid phase for a noncompetitive, one-step, kinetic immunoassay with varying incubation times and time-resolved fluorescence detection. RESULTS: The analytical (mean of zero + 3 SD) and functional (CV <15%) detection limits were 1.5 and 6.0 mIU/L for 2-min, 0.5 and 1.5 mIU/L for 7-min, and 0.2 and 0.5 mIU/L for 15-min assays, respectively. A good correlation was found with the Chiron Diagnostics ACS:180 assay (slopes 0.885-1.051, y-intercepts < +/- 0.20 mIU/L, S(y logical or, bar below x) 0.98, n = 20). CONCLUSION: The kinetic TSH assay provides reproducible and quantitative information on thyroid status within minutes and is applicable for the detection of hypothyroidism in point-of-care (POC) conditions. PMID- 12633762 TI - Evaluation of a fully-automated particle-enhanced turbidimetric immunoassay for the measurement of plasma lipoprotein(a). population-based reference values in an area with low incidence of cardiovascular disease. AB - OBJECTIVES: Lipoprotein(a) has been proposed as an independent risk factor for cardiovascular disease. This lipoprotein possesses a marked size polymorphism that makes difficult to measure accurately its concentration in plasma. The International Federation of Clinical Chemistry recently recommended to carefully evaluate new commercial methods for lipoprotein(a) measurement to discard the possible influence of lipoprotein(a) isoforms on immunoreactivity. They also recommended to perform population-based studies for different ethnic and geographic groups. Therefore, in the evaluation of a fully automated, particle enhanced turbidimetric immunoassay for the measurement of lipoprotein(a) we have determined its reference interval in the Spanish population, an area with the lowest incidence of cardiovascular disease in Europe. DESIGN AND METHOD: We evaluated a commercial kit of reagents calibrated against the Proposed Reference Material and determined the effect of lipoprotein(a) size polymorphism on the measurements. A population-based study was carried out in two different villages on the Mediterranean coast of Spain. RESULTS: Imprecision at different lipoprotein(a) concentrations ranged between 3.0 and 15.4%. Recovery was 98.5 +/- 2.1. Detection limit was 4.8 nmol/L. There were no significant interferences from lipemia, jaundice, hemolysis, paraproteinemia, apolipoprotein B or plasminogen. We did not observe any effect of the lipoprotein(a) size polymorphism on the measurements. Mean (and SD) values for plasma lipoprotein(a) (n = 369) were 53.6 (65.3) nmol/L, the median was = 25.3 nmol/L and range varied between <4.8 and 356.0 nmol/L. CONCLUSION: The present article presents an accurate and practical assay for measuring plasma lipoprotein(a) concentrations and describes its reference values in a population of Spanish Caucasians. Our results are similar to those obtained in other Caucasian populations (between 10 and 25% higher than in participants of the CARDIA study). PMID- 12633763 TI - Prognostic value of quantitatively assessed KLK7 expression in ovarian cancer. AB - BACKGROUND: Among females, ovarian cancer is the sixth most common malignancy. Women with ovarian cancer have poor overall survival rates, largely because the disease is often diagnosed at an advanced, less curable stage. Several lines of evidence suggest that members of the kallikrein family are involved in various malignancies such as prostate (PSA, KLK2, KLK15), ovarian (KLK4, KLK5, KLK6, KLK8, KLK10), and breast cancer (KLK10, KLK13, KLK14). Recent evidence has indicated that expression of KLK7 appears to be increased in ovarian cancer. We hypothesized that overexpression of the KLK7 gene in ovarian cancer may serve as a prognostic marker of the disease. METHODS: Using the LightCycler technology we quantified the level of KLK7 mRNA expression in 125 ovarian tumors. Different disease stages and tumor grades were analyzed. Univariate and multivariate analyses were performed to establish the associations between clinicopathological parameters and KLK7 expression. RESULTS: We here report that patients with KLK7 negative tumors have a significantly higher disease-free survival than patients with KLK7-positive tumors. KLK7 expression levels were significantly higher in patients with grade 3 than in patients with grade 1 to 2 tumors (p = 0.030). KLK7 status also correlated with size of residual tumor postsurgery. KLK7 expression is an independent predictor of both disease-free and overall survival for patients with low grade tumors. In this subgroup of patients the hazard ratios for disease-free and overall survival were 3.28 and 3.09, respectively. Similarly, patients who had undergone optimal debulking but harbored KLK7 positive tumors had a high hazard ratio (HR) for relapse (HR = 8.2) and death (HR = 4.6). CONCLUSIONS: We conclude that higher KLK7 expression in ovarian cancer tissue is associated with poorer prognosis of ovarian cancer patients, especially those with lower grade disease and those who have been optimally debulked. PMID- 12633764 TI - A genomic approach to mutation analysis of holocarboxylase synthetase gene in three Chinese patients with late-onset holocarboxylase synthetase deficiency. AB - OBJECTIVE: Multiple carboxylase deficiency (MCD, MIM:253270) is a common organic aciduria and caused by deficiency of either biotinidase or holocarboxylase synthetase (HLCS; EC 6.3.4.10). Patients commonly present during early infancy with acute metabolic derangements and severe metabolic acidosis. Recently, a late onset form of HLCS deficiency was also described. The different phenotypes (early and late presenting) may be related to a spectrum of mutations in HLCS gene. Applications of mutation analysis in HLCS had been limited previously by the requirement of cDNA from living tissue for study. We described here a genomic approach for molecular diagnosis of HLCS deficiency which we have used to detect mutations in Chinese patients who had the late-onset form of HLCS deficiency. In addition, a fibroblast cell line with MCD from Coriell Cell repositories was also studied. DESIGN AND METHODS: Three Chinese patients with late onset HLCS deficiency were studied. The genomic sequence of HLCS was retrieved and newly designed primers were used to cover all coding sequences of the gene. PCR products were analyzed by direct sequencing. Population allelic frequencies of mutations detected were determined by genotyping of control samples by restriction fragment length polymorphism. RESULTS: We found a recurrent mutation, R508W, in the three unrelated Chinese patients. Two were homozygous for this mutation. The other patient was a compound heterozygote of R508W and a novel mutation, D634N. The results suggest that R508W may be an important and relatively prevalent disease-causing mutation in Chinese MCD patients. A fibroblast cell-line from an African patient revealed an additional novel mutation, R565X and a known mutation, V550M. CONCLUSION: R508W is a recurrent mutation in Chinese MCD patients which is associated with the late onset phenotype. This new genomic approach for mutation analysis of HLCS gene provides new opportunities in studies of MCD. PMID- 12633765 TI - Improved diagnosis of a common mutation (R248C) in the human growth factor receptor 3 (FGFR3) gene that causes type I Thanatophoric dysplasia. PMID- 12633768 TI - The study of muscle action during single support and swing phase of gait: clinical relevance of forward simulation techniques. AB - Individual muscle function during single stance and swing phase of gait were analyzed using muscle driven forward simulation. The activation of each of 22 muscles in a musculoskeletal model with seven degrees of freedom were excluded from the forward simulation and the resulting changes in joint angles studied. A classification of muscle function during single support and swing phase of gait is presented. Altered joint kinematics due to the absence of individual muscle action is discussed in the light of pathological gait kinematics and clinical decision-making. PMID- 12633766 TI - A practical method to study functional impairment of proteins by glycation and effects of inhibitors using current coagulation/fibrinolysis reagent kits. AB - Objectives We undertook the present work to device a simple method to study the effects of inhibitors on functional impairment of proteins by the action of glycating agents. Design and methods For that purpose, we first tested the feasibility and optimized the conditions to employ glycation of human plasma coupled with AT III and plasminogen activity measurement, using coagulation test kits available in most clinical laboratories. Results Using D-BUT-CHT-lys-pNA as a plasmin-specific substrate, we show that incubation of plasma with fructose, glyceraldehyde or MG but not glucose decreases plasminogen activity reaching more than 40% in 16 h. A parallel dose-dependent decrease in heparin activation of AT III by up to a 50% was demonstrated using SAR-PRO-ARG-pNA as a specific thrombin substrate. We studied the effects of aminoguanidine, carnosine, quercetin aglycone, alpha tocopherol and ascorbic acid. Conclusion The methods afforded good discrimination between the known different reactivities of glycating sugars as well as the action of known antiglycation agents. They provide a practical system for monitoring the action of putative antiglycation agents. PMID- 12633769 TI - Predicting peak kinematic and kinetic parameters from gait speed. AB - OBJECTIVE: The aim of this study is to assess the predictability of the relationships between gait speed and common peak sagittal plane parameters in order to provide a set of reference parameter values. DESIGN: Lower extremity biomechanical data were collected in 64 healthy adults while walking barefoot at his/her comfortable walking speed, then at self-selected fast, slow and very slow speeds. Twenty seven peak joint parameter values were plotted and regressed as a function of gait speed. DISCUSSION: While most parameters change with increasing gait speed, in general, the kinetic parameters had better predictability than the kinematic parameters. Most of the power parameters were found to have a quadratic relationship with gait speed. Of the moment parameters, four had a linear relationship with gait speed, while four had a quadratic one. These relationships shown in the tables and graphs here can be used as a reference for 'normal' gait parameter values. PMID- 12633770 TI - The effect of tibial torsion on the dynamic function of the soleus during gait. AB - An induced acceleration analysis (IAA) model was used to investigate the effect of tibial torsion on the ability of the soleus to support and propel the body during gait. The IAA approach is well suited for this study because it allows soleus action to be quantified in the presence of varying amounts of tibial torsion, while other factors such as body configuration and muscle activation are held constant. The results of the analysis showed that excess tibial torsion shifts the induced knee joint accelerations toward flexion, valgus and external rotation, and diminishes body center-of-mass support and propulsion. This analysis supports the concept that bony mal-alignment can lead to 'lever arm dysfunction'. The objective data correlate with previous clinical observations related to valgus stress, crouch and the role of the soleus in level walking. The IAA model provides a tool for examining various aspects of abnormal gait independently and quantitatively. PMID- 12633771 TI - Accurate measurement of muscle belly length in the motion analysis laboratory: potential for the assessment of contracture. AB - Two-dimensional ultrasound imaging was combined with motion analysis technology to measure distances between remote anatomical landmarks. The length of the belly of the medial gastrocnemius muscle in five normal adults (nine limbs) was estimated using this technique. Our results in vivo were similar to the reported data for the lengths of muscles in cadavers, and were consistent with the expected relationship between muscle belly length and ankle joint angle. Experiments in vitro demonstrated that the accuracy of the device was better than 2 mm over 20 cm. Measurements on the same subject on different occasions showed that the results were repeatable in vivo. Rendering of the reconstructed volume of a foam phantom gave results comparable to photographic images. This validated technique could be used to measure muscle lengths in children with spastic cerebral palsy and indicate which muscles had fixed shortening, and to what extent. PMID- 12633772 TI - An in vitro study of individual ankle muscle actions on the center of pressure. AB - The objective of this study was to correlate the effects of muscle force on the movement of the center of pressure (COP) for increased clinical utility of the COP measurement. Five fresh frozen cadaveric specimens were used to apply a 49 N sinusoidal muscle force to isolated or grouped extrinsic ankle muscles, and a constant ankle joint reaction force at different tibial positions. The muscle force and the vertical ground reaction force (GRF) play the role of a mechanical lever system so that the differential COP movement can be interpreted as a moment arm for the vertical GRF. PMID- 12633773 TI - The spanning set indicates that variability during the stance period of running is affected by footwear. AB - Sensory information the foot receives appears to be related to kinematic variability. Since footwear material densities affect sensory information, footwear may be an important factor that dictates variability. This study hypothesized that modifications in footwear would result in changes in kinematic variability during the running stance period. Subjects ran on a treadmill for three conditions: hard shoe, soft shoe and barefoot. The spanning sets of the mean ensemble curves of the knee and ankle changes for each condition were used to define variability. Variability was significantly larger in the barefoot condition in comparison with the two footwear conditions for both joints. These results suggest that variability can be affected by peripheral sensory information. The spanning set methodology can be utilized to examine changes in variability. PMID- 12633774 TI - Dynamic and static tests of balance and postural sway in boys: effects of previous wrist bone fractures and high adiposity. AB - Ninety-three males aged 10-21 years undertook the Bruininks-Oseretsky balance test and two computerized posturography tests to evaluate the effects of (a) previous forearm fracture and (b) high body weight on balance and postural sway. Body composition was measured by dual energy X-ray absorptiometry. Fracture history did not affect balance measures. However, Bruininks-Oseretsky balance scores were negatively correlated with body weight, body mass index, percentage fat and total fat mass. Overweight subjects (n=25) had lower scores (P<0.05) than boys of healthy weight (n=47), supporting the view that overweight adolescents have poorer balance than those of healthy weight. PMID- 12633775 TI - Adjustments in gait symmetry with walking speed in trans-femoral and trans-tibial amputees. AB - The effect of increased walking speed on temporal and loading asymmetry was investigated in highly active trans-femoral and trans-tibial amputees. With increasing walking speed, temporal gait variables reduced in duration, particularly on the prosthetic limb, while vertical ground reaction force (vGRF) increased in magnitude, particularly on the intact limb. Thus, temporal asymmetry reduced and loading asymmetry increased with walking speed. The greater force on the intact limb may reflect the method by which the amputees achieve greater temporal symmetry in order to walk fast, and could possibly account for greater instances of joint degeneration in the intact limb reported in the literature. PMID- 12633776 TI - The influence of external orthotic support on the adaptive gait characteristics of individuals with chronically unstable ankles. AB - External orthotic supports or braces are used clinically to treat individuals with chronically unstable ankles. Braces have efficacious effects on dynamic strength, passive tissue tension, and proprioception, but their effects on gait remains undetermined. The purpose of this study was to determine if overground locomotion, including walking up a step and a curb are influenced by orthotic device use. Twenty subjects, 10 with no previous injury and 10 with chronically unstable ankles, performed with no brace, a flexible brace and a semi-rigid brace while walking on a level surface, up a step and on a ramp. Differences were noted in kinematic and kinetic gait parameters between groups and between step and ramp conditions, suggesting that gait patterns vary between individuals who have had ankle sprain and those who have not. Few differences were noted between the brace and no brace conditions for both groups. Adding to the previously described efficacious benefits, we conclude that bracing does not alter selected gait parameters in individuals who have chronic ankle instability. PMID- 12633777 TI - Individual muscle contributions to support in normal walking. AB - The purpose of this study was to quantify the contributions made by individual muscles to support of the whole body during normal gait. A muscle's contribution to support was described by its contribution to the time history of the vertical force exerted by the ground. The analysis was based on a three-dimensional, muscle-actuated model of the body and a dynamic optimization solution for normal walking. The results showed that, in early stance, before the foot was placed flat on the ground, support was provided mainly by the ankle dorsiflexors. After foot-flat, but before contralateral toe-off, support was generated primarily by gluteus maximus, vasti, and posterior gluteus medius/minimus; these muscles were responsible for the first peak seen in the vertical ground-reaction force. The majority of support in midstance was provided by gluteus medius/minimus, with gravity assisting significantly as well. The ankle plantarflexors generated nearly all support in late stance; these muscles were responsible for the second peak in the vertical ground-reaction force. The results showed also that centrifugal forces act to decrease the vertical ground-reaction force, but only by minor amounts, and that resistance of the skeleton to the force of gravity is no larger than 1/2 body weight throughout the gait cycle. PMID- 12633778 TI - Kinetic analysis of planned gait termination in healthy subjects and patients with balance disorders. AB - We examined the stability and strategies used by balance impaired (vestibular hypofunction, VH: n=25; cerebellar damage, CB: n=20) patients and healthy (HE: n=52) controls during planned gait termination. Upper body strategies (during final stride and final step) were investigated using peak positive kinetic power (KP((+)): kinetic energy increasing), and peak negative kinetic power (KP((-)): kinetic energy decreasing) of the head-arms-trunk segment. Compared to HE controls (P<0.05): CB patients' medio-lateral KP((+)) and KP((-)) and were 53 and 71% higher during final stride, respectively; VH patients medio-lateral KP((+)) and KP((-)) was 78 and 57% higher during final step, respectively, and; during the final, standing stage VH patients were 32% less stable (from phase plane analysis) in the frontal plane. The excessive energy transfers in final stride for CB patients was likely due to poor eccentric muscle control when preparing for the stop. VH patients had difficulty controlling lateral stability during final step and once they had stopped walking, probably due to the lack of vestibular feedback regarding forward velocity changes. A better understanding of these abnormal movement patterns or compensatory strategies may assist in rehabilitation of patients with balance dysfunction. PMID- 12633779 TI - Effect of foot orthoses on the kinematics and kinetics of normal walking gait. AB - Despite their wide clinical application and success, our understanding of the biomechanical effects of foot orthoses is relatively limited. The aim of this study was to assess the effect of medially wedged and laterally wedged foot orthoses on the kinematics and joint moments of the rearfoot complex, knee, hip and pelvis and the ground reaction forces. The principal effect of the foot orthoses was on the rearfoot complex, where significant changes in joint rotations and moments were observed. Medially wedged orthoses decreased rearfoot pronation and increased the laterally directed ground reaction force during the contact phase, suggesting reduced shock attenuation. The laterally wedged orthoses increased rearfoot pronation and decreased the laterally directed ground reaction force during the contact phase, suggesting increased shock attenuation. The effects of the orthoses on knee, hip and pelvis kinematics were generally minimal. In view of the minimal effect the orthoses had on joints proximal to the foot, it is suggested that the orthoses may have additional effects on the passive and active soft tissues of the lower limb and it is these changes that result in the documented clinical success. PMID- 12633782 TI - Noggin arrests stromal cell differentiation in vitro. AB - Noggin is a glycoprotein that binds bone morphogenetic proteins (BMPs) selectively and, when added to osteoblasts, it opposes the effects of BMPs. However, the consequences of its continued expression in stromal cells are not known. We investigated the effects of noggin overexpression under the control of a constitutive promoter, on murine ST-2 stromal cells, and its impact on stromal cells from transgenic mice overexpressing noggin under the control of the osteocalcin promoter. ST-2 cells were transduced with a retroviral vector (pLPCX) or a vector driving noggin (pLPCX noggin). Untreated (pLPCX) ST-2 cells developed the appearance of mineralized nodules and expressed osteocalcin. pLPCX noggin delayed the appearance of mineralized nodules and prevented the expression of osteocalcin. Noggin also prevented the cortisol-dependent induction of peroxisome proliferator-activated receptor gamma2 and adipsin transcripts, indicating a generalized inhibition of cell differentiation. Primary stromal cells from noggin transgenic mice displayed impaired differentiation when compared to cells from wild-type animals and did not express osteocalcin mRNA. In conclusion, noggin arrests the differentiation of stromal cells, preventing cellular maturation. PMID- 12633783 TI - Long-term efficacy of risedronate: a 5-year placebo-controlled clinical experience. AB - Limited placebo-controlled data are available to assess the long-term fracture efficacy of bisphosphonates. In order to determine the effects of 5 years of risedronate treatment, we extended a 3-year, placebo-controlled vertebral fracture study in osteoporotic women for an additional 2 years; women who entered the extension study continued to receive 5 mg risedronate or placebo according to the original randomization, with maintenance of blinding. End points included vertebral and nonvertebral fracture assessments, bone mineral density measurements, and changes in biochemical markers of bone turnover. A total of 265 women (placebo, 130; 5 mg risedronate, 135) entered the study extension and 220 (83%) completed the additional 2 years. Fracture results observed in the study extension were consistent with those observed in the first 3 years. The risk of new vertebral fractures was significantly reduced with risedronate treatment in years 4 and 5 by 59% (95% confidence interval, 19 to 79%, P = 0.01) compared with a 49% reduction in the first 3 years. Rapid and significant decreases in markers of bone turnover observed in the first 3 years were similarly maintained in the next 2 years of treatment. Increases in spine and hip bone mineral density that occurred in the risedronate group during the first 3 years were maintained or increased with a further 2 years of treatment. The mean increase from baseline in lumbar spine BMD over 5 years was 9.3% (P < 0.001). This study demonstrates that the effects of risedronate over 3 years on vertebral fracture and BMD are maintained with a further 2 years of treatment. PMID- 12633784 TI - Orthosilicic acid stimulates collagen type 1 synthesis and osteoblastic differentiation in human osteoblast-like cells in vitro. AB - Silicon deficiency in animals leads to bone defects. This element may therefore play an important role in bone metabolism. Silicon is absorbed from the diet as orthosilicic acid and concentrations in plasma are 5-20 microM. The in vitro effects of orthosilicic acid (0-50 microM) on collagen type 1 synthesis was investigated using the human osteosarcoma cell line (MG-63), primary osteoblast like cells derived from human bone marrow stromal cells, and an immortalized human early osteoblastic cell line (HCC1). Collagen type 1 mRNA expression and prolyl hydroxylase activity were also determined in the MG-63 cells. Alkaline phosphatase and osteocalcin (osteoblastic differentiation) were assessed both at the protein and the mRNA level in MG-63 cells treated with orthosilicic acid. Collagen type 1 synthesis increased in all treated cells at orthosilicic acid concentrations of 10 and 20 microM, although the effects were more marked in the clonal cell lines (MG-63, HCCl 1.75- and 1.8-fold, respectively, P < 0.001, compared to 1.45-fold in the primary cell lines). Treatment at 50 microM resulted in a smaller increase in collagen type 1 synthesis (MG-63 1.45-fold, P = 0.004). The effect of orthosilicic acid was abolished in the presence of prolyl hydroxylase inhibitors. No change in collagen type 1 mRNA level was seen in treated MG-63 cells. Alkaline phosphatase activity and osteocalcin were significantly increased (1.5, 1.2-fold at concentrations of 10 and 20 microM, respectively, P < 0.05). Gene expression of alkaline phosphatase and osteocalcin also increased significantly following treatment. In conclusion, orthosilicic acid at physiological concentrations stimulates collagen type 1 synthesis in human osteoblast-like cells and enhances osteoblastic differentiation. PMID- 12633785 TI - The effects of estrogen on osteoprotegerin, RANKL, and estrogen receptor expression in human osteoblasts. AB - Estrogen is essential for bone growth and development and for the maintenance of bone health in adulthood. The cellular responses of osteoblasts and osteoclasts to estrogen are initiated via two high-affinity receptors (ERs). Osteoblasts synthesize RANKL (receptor activator of NF-kappaB ligand), necessary for osteoclast formation and function, and osteoprotegerin (OPG), its decoy receptor. To investigate the effects of estrogen on the expression of OPG, RANKL, and ERs in human osteoblasts, cells were cultured with physiological (10(-10) M) and high dose (10(-7) M) 17beta-estradiol for 24 and 48 h. Proteins and corresponding mRNA levels were quantitatively determined by immunocytochemistry and RT-PCR. OPG expression was significantly increased three- and sevenfold at 24 h with 10(-10) M (P < 0.05) and 10(-7) M (P < 0.01) estradiol, respectively, compared to untreated cells. Similar but smaller increases were seen at 48 h (P < 0.05). Osteoblasts treated with estradiol demonstrated increased RANKL protein expression at 24 h (P < 0.05), but this was not maintained at 48 h. ERalpha expression was significantly increased by high-dose estradiol (P < 0.01) at 24 h and dose-dependently increased at 48 h (P < 0.01), while ERbeta was only increased at 24 h (P < 0.01). The estrogen-induced protein expression of ER, OPG, and RANKL was abrogated when cells were cultured in the presence of the estrogen antagonist ICI 182780. mRNA levels at 24 h demonstrated a significant suppression of RANKL with the low-dose but not the high dose. ERalpha mRNA but not ERbeta expression was up-regulated by estrogen. Our results suggest that estrogen may exert its anti-resorptive effects on bone, at least in part, by stimulating ER and OPG expression in osteoblasts. PMID- 12633787 TI - Organization of apatite crystals in human woven bone. AB - The organization of collagen fibrils differs in woven bone and lamellar bone, and it reflects certain aspects of the nature of the mineral crystals associated with them. In order to investigate the morphology and distribution of apatite crystals in woven bone, mineralized collagen fibrils and isolated crystals from the mid diaphyses of human fetal femurs were observed with scanning and transmission electron microscopy and high-resolution electron microscopy. A number of features of woven bone were observed for the first time by these means. Similar to mature crystals from lamellar bone, the apatite crystals in woven bone are also platelet shaped. However, most likely because of a high rate of old bone resorption and new bone formation in woven material, the average crystal dimensions are considerably smaller than those of mature crystals in lamellar bone. Apatite crystals were noted on the surface of collagen fibrils in woven bone. In densely packed woven bone, the periodicity of mineral deposited on individual fibrils is in registration over many fibrils. In addition to their association with collagen surfaces, crystals also appear distributed in both extrafibrillar and intrafibrillar collagen regions. In both cases, the minerals are crystalline and defect-free. These characteristics provide insight into the spatial and temporal relation between collagen and mineral that is the basis for the structure and organization of the mineral comprising human woven bone. PMID- 12633786 TI - Effect of nitric oxide donor nitroglycerin on bone mineral density in a rat model of estrogen deficiency-induced osteopenia. AB - Nitric oxide (NO) may modulate estrogen's anabolic effects on bone homeostasis by restraining osteoclast-mediated bone resorption and stimulation of osteoblast activity. Accordingly, NO donated by organic nitrates, including nitroglycerin, is thought to protect against bone loss associated with estrogen deficiency. In this study, we have explored this phenomenon. Thirty-two 12-week-old female Wistar rats were divided into four groups prior to bilateral ovariectomy or a sham operation. The ovariectomised rats received (1). vehicle control (OVX control), (2). 17-beta-estradiol (OVX+E2), or (3). transdermal nitroglycerin (OVX+NG) for 4 weeks. Femoral and tibial bone mineral density (BMD), serum alkaline phosphatase and urine deoxypyridinoline and NO metabolites were analysed at the end of the study period together with failure torque and torsional rigidity of the tibiae and cellular localisation of the NO-synthase (NOS) isoforms. In OVX+E2 group, proximal and distal femoral and proximal tibial BMD exceeded that of the Sham controls. Nitroglycerin prevented BMD loss at these three sites at levels comparable to that of the Sham controls. Deoxypyridinoline excretion did not change except in the OVX-E2 group that showed an expected reduction when compared to the Sham and OVX controls. There were no treatment related differences in total alkaline phosphatase or urinary NO metabolites. Tibial failure torque was comparable between the groups but both OVX+E2 and OVX+NG groups showed decreased torsional rigidity compared with the OVX controls. Endothelial and inducible NOS were found in osteoblast-like cells associated with calcifying cartilage spicules in the distal femoral metaphysis. These data confirm previous findings and show that nitroglycerin counteracts the estrogen deficiency-induced osteopenia in the ovariectomised rat model. Organic nitrates may thus be beneficial in conditions where bone turnover is compromised such as in osteoporosis. PMID- 12633788 TI - Trabecular minimodeling in human iliac bone. AB - In adult human beings, remodeling creates nearly all of new bone tissue. However, Frost hypothesized that modeling can go on in trabeculae throughout life. As this hypothesis has not been verified, we looked for histologic evidence of trabecular modeling (minimodeling) during bone histomorphometry of transiliac bone biopsy specimens obtained from 34 patients (age range, 38-81 years; mean age, 58.4 years; female, 31/34) at the time of total hip arthroplasty. Before the bone biopsy study, we performed quantitative bone scintigraphy of bilateral hip joints and bilateral iliac crests in 10 other patients with unilateral hip disease and confirmed that the bone biopsy site was not affected by ipsilateral hip joint disease. Patients who had metabolic bone diseases or who had taken medications known to affect bone metabolism were excluded from the study. During modeling where bone formation and bone resorption are not coupled, bone formation can occur on quiescent bone surfaces without preceding bone resorption and create smooth cement lines. Therefore, the combination of fluorochrome labeling and a smooth cement line without interruption of surrounding collagen fibers was regarded as evidence of minimodeling. Histologic evidence of minimodeling was detected in 21 of the entire 34 specimens (62%) and 17 of 27 specimens obtained from postmenopausal patients (63%). Bone volume of minimodeling sites was less than 1% of the trabecular bone volume, and these sites accounted for less than 2% of the entire bone surface on average. However, osteoid volume of minimodeling sites comprised approximately one-tenth of the entire osteoid volume, and their labeled surface constituted one-fourth to half of the entire labeled surface on average. Therefore, when performing bone histomorphometry of adult cancellous bone, minimodeling should be taken into account when dealing with parameters related to osteoid volume and mineralization. A comparison of specimens with and without minimodeling demonstrated that the presence of minimodeling was correlated with smaller physique of patients, accelerated mineralization (as indicated by the higher mean MS/BS and MAR values and the shorter mean Omt), and higher metabolic turn-over of bone (as indicated by the higher mean BFR/BV value). Although the findings still need to be verified in a larger number of normal subjects without hip joint disease, they support Frost's hypothesis that minimodeling can continue throughout human life. PMID- 12633789 TI - Parathyroid hormone secretory pattern, circulating activity, and effect on bone turnover in adult growth hormone deficiency. AB - Adult growth hormone deficiency (AGHD) is associated with osteoporosis. Reports have associated parathyroid hormone (PTH) circadian rhythm abnormalities with osteoporosis. Furthermore, there is evidence of relative PTH insensitivity in AGHD patients. Factors regulating PTH circadian rhythm are not fully understood. There is evidence that serum phosphate is a likely determinant of PTH rhythm. The aim of this study was to investigate PTH circadian rhythm and its circulating activity and association with bone turnover in untreated AGHD patients compared to healthy individuals. We sampled peripheral venous blood at 30-min and urine at 3-h intervals during the day over a 24-h period from 1400 h in 14 untreated AGHD patients (7 M, 7 W; mean age, 49.5 +/- 10.7 years) and 14 age (48.6 +/- 11.4 years; P = NS) and gender-matched controls. Cosinor analysis was performed to analyze rhythm parameters. Cross-correlational analysis was used to determine the relationship between variables. Serum PTH (1-84), phosphate, total calcium, urea, creatinine, albumin, type I collagen C-telopeptides (CT(x)), a bone resorption marker, and procollagen type I amino-terminal propeptide (PINP), a bone formation marker, were measured on all samples. Nephrogenous cyclic adenosine monophosphate (NcAMP), which reflects the renal activity of PTH, was calculated from plasma and urinary cAMP. Urinary calcium and phosphate were measured on all urine samples. Significant circadian rhythms were observed for serum PTH, phosphate, CT(x), and PINP in AGHD and healthy subjects (P < 0.001). No significant rhythm was observed for serum-adjusted calcium. PTH MESOR (rhythm-adjusted mean) was significantly higher (P < 0.05), whereas the MESOR values for phosphate, CT(x) (P < 0.05), and PINP (P < 0.001) were lower in AGHD patients than in controls. AGHD patients had significantly lower 24-h NcAMP (P < 0.001) and higher urinary calcium excretion (P < 0.05). Maximum cross-correlation between PTH and phosphate (r = 0.75) was observed when PTH was lagged by 1.5 h in healthy individuals, suggesting that changes in phosphate precede changes in PTH concentration. PTH/CT(x) and PTH/PINP showed maximum correlation when CT(x) (r = 0.68) and PINP (r = 0.71) were lagged by 3 h. In AGHD patients, compared to controls the maximum correlation between PTH/phosphate (r = 0.88, P = 0.007), PTH/CTx (r = 0.61, P = 0.027), and PTH/PINP (r = 0.65, P = 0.028) was observed when the lag time was reduced by 1.5 h in all variables, with changes in PTH and phosphate occurring at concurrent time points. Our data suggest decreased end-organ sensitivity to the effects of PTH in AGHD patients, resulting in a significantly lower NcAMP, low bone turnover, and higher calcium excretion in the presence of significantly higher PTH concentrations. We have also demonstrated that changes in serum phosphate precede those of PTH, which in turn precede changes in bone resorption and formation in healthy individuals. This relationship was altered in AGHD patients. These results suggest a possible role for GH in regulating PTH secretion and the bone remodeling process. PMID- 12633790 TI - Hip fracture patients have more vertebral deformities than subjects in population based studies. AB - The prevalence of vertebral deformity, estimated in lateral spine radiographs (Th4-L4) using quantitative morphometry, in 64 men and 132 women with hip fractures was compared with the prevalence of vertebral deformity in individuals in two population-based studies. A vertebral deformity of a specific vertebra was defined as a 3, 4, or 5 SD reduction from the normal mean of any of three ratios describing that specific vertebra. The age-adjusted prevalence of individuals with vertebral deformities was higher among the hip fracture patients than among the reference subjects in both genders, with an odds ratio of 3.6 [95% confidence interval (CI) 1.9-6.6] in men and 2.6 (95% CI 1.7-4.1) in women using deformity criterion -3 SD. Also, the number of vertebrae with deformities (-3 SD) in individuals with one deformity or more was greater among the hip fracture patients than among the reference subjects (in men mean 2.3 versus 1.8, P = 0.007; in women mean 3.3 versus 2.0, P < 0.001). Adjusted for age there were more vertebrae with deformities (-3 SD) among female than among male hip fracture patients (mean 3.3 versus 2.3, P = 0.01). We found no differences in the vertebral deformity rates when comparing patients with a per- or subtrochanteric hip fracture with patients with a cervical hip fracture. In conclusion, there is a relationship between vertebral deformities and hip fractures suggesting that a prevalent vertebral deformity could predict an increased hip fracture risk. PMID- 12633791 TI - Association of a Trp16Ser variation in the gonadotropin releasing hormone signal peptide with bone mineral density, revealed by SNP-dependent PCR typing. AB - Osteoporosis is believed to result from interplay among multiple environmental and genetic determinants, including factors that regulate bone mineral density (BMD). Among those factors, adequate estrogen is essential for achievement of peak bone mass as well as for postmenopausal maintenance of skeletal homeostasis. Gonadotropin-releasing hormone (GnRH) from the hypothalamus is the primary determinant in the hypothalamic-pituitary-gonadal feedback system. In genetic studies of 384 postmenopausal Japanese women, we found a significant association between BMD and an amino acid variation (Trp16Ser) located within the signal peptide of GnRH (r = 0.143, P = 0.005). These results were achieved by genotyping all subjects using a newly developed SNP-dependent PCR method. This automated, high-throughput, and inexpensive procedure is suitable for typing large numbers of samples. BMD was lowest among 16Ser/Ser homozygotes, highest among 16Trp/Trp homozygotes, and intermediate among heterozygotes. A case-control study involving 125 osteoporosis patients and 92 healthy controls revealed a significant association between the presence of a 16Ser GnRH allele and affected status (chi(2) = 4.74, P = 0.041). The results suggested that variation of the GnRH signal peptide may be an important risk factor for postmenopausal osteoporosis. PMID- 12633792 TI - Quantitative genetic study of radiographic hand bone size and geometry. AB - Despite the obvious epidemiological significance of bone size (BS) and geometry (BG) traits as risk factors for osteoporotic fracture, very little is still known concerning the extent of their genetic determination. In the present paper we report the results of quantitative genetic analysis of a number of BG and BS indices, as well as of BMD measurements, obtained on a large pedigree-based sample (296 nuclear families, 1208 individuals) of plain hand radiographs. The families studied were all ethnically Caucasians (Chuvasha) living in small villages along the Volga River (Russia). The sample consisted of 636 men and 572 women, aged 18-91 years. To assess hand bone size we used the outcome of principal component analysis conducted on 48 measurements of metacarpal bones and proximal phalanges (PC-BS). Two BG indices, average metacarpal cortical index and breaking bending resistance index (BBRI), also measured on metacarpal and proximal phalanges were used. Again the outcome of the principal component PC BBRI was examined in the genetic analysis. PC-BS measurements strongly correlated with body length (r = 0.75, P < 0.001) and weight (r = 0.39, P < 0.001), suggesting that they indeed reflected hand skeleton size. Familial correlations for all studied traits, adjusted for covariates (sex, age, etc.), were all highly significant statistically. For example, parent/offspring correlations ranged between 0.248 (P < 0.001) for phalangeal BMD and 0.385 (P < 0.001) for PC-BBRI. Maximum likelihood estimates of the variance component analysis confirmed these results, indicating that approximately 58 to 66% of the residual variance of the studied traits was attributable to genetic effects. Bivariate analysis clearly revealed that while genetic variation of the phalangeal BMD was independent of the genetic effects influencing hand BS and BG, the latter two were strongly interrelated. A substantial proportion of PC-BS and PC-BBRI variation was due to shared genetic (r(G) = 0.468 +/- 0.063) and environmental (r(E) = 0.704 +/- 0.052) factors. PMID- 12633793 TI - Effects of prior beta-blocker therapy on clinical outcomes after primary coronary angioplasty for acute myocardial infarction. AB - We hypothesized that pretreatment with beta blockers may improve clinical outcomes after primary angioplasty for acute myocardial infarction. We pooled clinical, angiographic, and outcomes data on 2,537 patients enrolled in the Primary Angioplasty in Myocardial Infarction (PAMI), PAMI-2, and Stent PAMI trials. We classified patients into a beta group (n = 1,132) if they received beta-blocker therapy before primary angioplasty or a no-beta group (n = 1,405) if they did not. We evaluated procedural complications and in-hospital and 1-year outcomes (death and major adverse cardiac events [death, reinfarction, target vessel revascularization, or stroke]) between groups. Beta patients were younger, had higher systolic blood pressure and heart rate, and were more likely to be in Killip class I at admission. They had lower left ventricular ejection fraction, greater door-to-balloon time, greater likelihood of having a left anterior descending artery culprit lesion, but a similar incidence of Thrombolysis In Myocardial Infarction 3 flow after angioplasty (92.6% vs 92.7%, p = 0.91). The beta group had less procedural complications (23% vs 34%, p <0.0001) and a lower incidence of death (1.8% vs 3.7%, p = 0.0035) and major adverse cardiac events (5.5% vs 7.8%, p = 0.027) during hospitalization. At 1 year, mortality remained lower in beta patients (4.9% vs 6.7%, log-rank p = 0.055). After adjustment for baseline differences, beta patients had significantly lower in-hospital mortality (odds ratio 0.41; 95% confidence interval 0.20 to 0.84; p <0.0148) and nonsignificantly lower 1-year mortality (odds ratio 0.72; 95% confidence interval 0.47 to 1.08; p = 0.11). Thus, pretreatment with beta blockers has an independent beneficial effect on short-term clinical outcomes in patients undergoing primary angioplasty for acute myocardial infarction. PMID- 12633794 TI - Percutaneous myocardial laser revascularization in patients with refractory angina pectoris. AB - This study aimed to determine the safety and efficacy of percutaneous myocardial laser revascularization (PMLR). Seventy-three patients with stable angina pectoris (class III or IV) who were unsuitable for conventional revascularization and had evidence of reversible ischemia by thallium-201 scintigraphy, ejection fraction of > or =25%, and myocardial wall thickness > or =8 mm were randomized to optimal medical therapy alone (n = 37) or PMLR with optimal medical therapy (n = 36). Patients were followed up at 3, 6, and 12 months. The primary end point was exercise time. Secondary end points included angina scores, left ventricular ejection fraction, quality of life, changes in medical therapy, and hospitalizations. All 36 patients randomized to PMLR underwent the procedure successfully with no periprocedure deaths. One patient developed sustained ventricular tachycardia that required electrical cardioversion, and 1 patient developed cardiac tamponade that required surgical drainage. At 12 months, exercise times improved by 109 seconds in the PMLR group but decreased by 62 seconds in the control group (p <0.01). Angina scores improved by 2 classes in 36% of PMLR-treated patients at 12 months compared with 0% of the control patients (p <0.01). We conclude that PMLR is a relatively safe procedure that provides patients with symptomatic angina relief and improvement in exercise capacity and quality of life. PMID- 12633795 TI - Comparison of once-daily, niacin extended-release/lovastatin with standard doses of atorvastatin and simvastatin (the ADvicor Versus Other Cholesterol-Modulating Agents Trial Evaluation [ADVOCATE]). AB - This study compared the relative efficacy of a once-daily niacin extended-release (ER)/lovastatin fixed-dose combination with standard doses of atorvastatin or simvastatin, with a special emphasis on relative starting doses. Subjects (n = 315) with elevated low-density lipoprotein (LDL) cholesterol and decreased high density lipoprotein (HDL) cholesterol blood levels (defined as LDL cholesterol blood levels > or =160 mg/dl without coronary artery disease, or > or =130 mg/dl if coronary artery disease was present, and HDL cholesterol <45 mg/dl in men and <50 mg/dl in women) were randomized to atorvastatin, simvastatin, or niacin ER/lovastatin for 16 weeks. The primary efficacy variables were the mean percent change in LDL cholesterol and HDL cholesterol levels from baseline. After 8 weeks, the starting dose niacin ER/lovastatin 1,000/40 mg and the 10-mg starting dose atorvastatin both lowered mean LDL cholesterol by 38%. After 12 weeks, niacin ER/lovastatin 1,000/40 mg lowered LDL cholesterol by 42% versus 34% with the 20-mg starting dose of simvastatin (p <0.001). Niacin ER/lovastatin increased HDL cholesterol significantly more than atorvastatin or simvastatin at all compared doses (p <0.001). Niacin ER/lovastatin also provided significant improvements in triglycerides, lipoprotein(a), apolipoprotein A-1, apolipoprotein B, and HDL subfractions. A total of 6% of study subjects receiving niacin ER/lovastatin withdrew because of flushing. No significant differences were seen among study groups in discontinuance due to elevated liver enzymes. No drug induced myopathy was observed. Niacin ER/lovastatin was comparable to atorvastatin 10 mg and more effective than simvastatin 20 mg in reducing LDL cholesterol, was more effective in increasing HDL cholesterol than either atorvastatin or simvastatin, and provided greater global improvements in non-HDL cholesterol, triglycerides, and lipoprotein(a). PMID- 12633796 TI - Effects of operator experience on the outcome and duration of pulmonary vein isolation procedures for atrial fibrillation. AB - Segmental, ostial ablation to isolate the pulmonary veins has been shown to be effective for the treatment of atrial fibrillation (AF). The purpose of this study was to determine the effects of operator experience on the outcome and duration of pulmonary vein isolation procedures for AF. One hundred three consecutive patients with AF underwent segmental, ostial ablation to isolate pulmonary veins. The effect of operator experience on the outcome and duration of the procedure was measured. The mean procedure duration was 232 +/- 70 minutes (range 50 to 460). There was an indirect linear relation between the total procedure time and the number of procedures performed (r = -0.68; p <0.001), and between the total fluoroscopic time and the number of procedures performed (r = 0.71; p <0.001). The percentage of patients in whom complete isolation of the pulmonary veins was achieved and the total duration of radiofrequency energy delivered during the procedures remained stable as operator experience increased. Operator experience was not an independent predictor of recurrent AF. The primary effect of operator experience on pulmonary vein isolation procedures for AF was a marked reduction in the duration of the procedure and fluoroscopic time. Long procedure durations and large amounts of radiation exposure may currently limit the widespread application of this therapy. The results of this study suggest that after >75 cases, segmental ablation to isolate the pulmonary veins using fluoroscopy and conventional radiofrequency ablation usually is feasible in <3 hours. PMID- 12633797 TI - Comparison of expression of connexin in right atrial myocardium in patients with chronic atrial fibrillation versus those in sinus rhythm. AB - An abnormal distribution of the gap junction occurs in chronic atrial fibrillation (AF). There are conflicting data regarding changes in connexins (Cxs) in experimental models of AF. We examined whether patients with chronic AF have alterations in atrial Cxs. We analyzed the expression of Cx40 and Cx43 in the right atrial myocardium from 10 patients with mitral valvular disease (MVD) who had AF (MVD/AF), 10 patients with MVD who were in normal sinus rhythm (MVD/NSR), and 10 control patients in NSR (tissue obtained during coronary artery bypass surgery). Hemodynamic and echocardiographic data were obtained before surgery, and an electrophysiologic examination was performed during the operation. An immunohistochemical study was performed on atrial tissue. The relative expression level of Cx40 protein was significantly lower in MVD/AF patients (6.5 +/- 4.6) than in either MVD/NSR patients (17.7 +/- 8.9, p <0.05) or controls (24.7 +/- 11.1, p <0.01). The relative expression level of Cx40 messenger ribonucleic acid was also significantly lower in MVD/AF patients (0.23 +/- 0.13) than in MVD/NSR patients (0.47 +/- 0.26, p <0.01) or controls (0.47 +/- 0.17, p <0.01). For Cx43 protein and messenger ribonucleic acid, there was no significant difference in relative expression levels among the 3 groups. Interestingly, the level of serine-phosphorylated Cx40 was approximately 52% greater in MVD/AF patients than in controls. In MVD/AF patients, the immunoreactive signal of Cx40 was significantly lower than in controls. There was no significant difference in the connective tissue-volume fraction among the groups. Thus, downregulation of Cx40 and abnormal phosphorylation of Cx40 may result in abnormal cell-to-cell communication and alteration in the electrophysiologic properties of the atrium, leading to the initiation and/or perpetuation of AF. PMID- 12633798 TI - Predictors of left ventricular reverse remodeling after cardiac resynchronization therapy for heart failure secondary to idiopathic dilated or ischemic cardiomyopathy. AB - Biventricular pacing results in left ventricular (LV) reverse remodeling in heart failure patients with wide QRS complexes. This study examines potential predictors of reverse remodeling. Echocardiography with tissue Doppler imaging was performed at baseline and 3 months after biventricular pacing in 30 patients (21 men and 9 women, mean age 62 +/- 14 years). There were 17 responders to reverse remodeling (defined as a reduction in LV end-systolic volume by >15%) and 13 nonresponders. Responders had significant improvement in 6-minute hall-walking distance (p = 0.006), metabolic equivalents (p = 0.02), peak oxygen uptake (p = 0.02), New York Heart Association functional class (p <0.001), and quality of life (p <0.001); an increase in the sphericity index (p = 0.007), ejection fraction (p <0.001), and diastolic filling time (p = 0.03); a decrease in myocardial performance index (p = 0.02), isovolumic relaxation time (p = 0.004), and mitral regurgitation (p = 0.007); and an improvement in systolic dyssynchrony (SD of the time to peak myocardial systolic contraction of the 12 LV segments as dyssynchrony index) (45.0 +/- 8.3 vs 32.5 +/- 14.5 ms, p = 0.003). In contrast, nonresponders only had a small degree of clinical improvement in New York Heart Association class (p = 0.03) and quality-of-life scores (p = 0.03), without any change in cardiac function, and worsening of systolic dyssynchrony (24.8 +/- 4.5 vs 34.1 +/- 13.5 ms, p = 0.02). When all the above factors were put into univariate and multivariate analyses models, systolic dyssynchrony was the only independent predictor of reverse remodeling (r = -0.76, p <0.001) (beta = -1.54, p = 0.007). A preimplant dyssynchrony index of 32.6 ms (+2 SDs from mean of 88 normal controls) was able to totally segregate responders from nonresponders of biventricular pacing. Thus, responders of LV reverse remodeling were associated with improvement in clinical status, cardiac function, and systolic synchronicity. Direct assessment of systolic synchronicity by tissue Doppler imaging is highly accurate in predicting responders to therapy. PMID- 12633799 TI - Effect of infective endocarditis on blood coagulation and platelet activation and comparison of patients with to those without embolic events. AB - Inflammation-induced procoagulant changes and alterations in platelet activity appear to play an important role in thromboembolic complications of infective endocarditis (IE). The aim of this study was to investigate systemic coagulation activity, fibrinolytic capacity, and platelet activation in IE patients with and without embolic events by measuring the plasma levels of prothrombin fragment 1 + 2, thrombin-antithrombin III complex, plasminogen activator inhibitor-1, beta thromboglobulin, and platelet factor 4. The study included 76 consecutive patients with definite IE according to the Duke criteria. Among them, 13 (17.1%) had major embolic events. Plasma concentrations of prothrombin fragment 1 + 2 (3.2 +/- 1.3 vs 1.7 +/- 0.7 and 1.4 +/- 0.7 nmol/L, p <0.001, respectively) and thrombin-antithrombin (7.3 +/- 1.5 vs 2.9 +/- 1.2 and 2.2 +/- 1.1 ng/ml, p <0.001, respectively) were elevated in patients with embolic events compared with both patients without embolic events and control subjects. Similarly, patients with embolic events had increased plasma levels of beta-thromboglobulin (63.3 +/- 10.9 vs 33.1 +/- 11.6 and 19.1 +/- 10.6 ng/ml, p <0.001, respectively) and platelet factor 4 (106.0 +/- 28.7 vs 50.3 +/- 16.7 and 43.0 +/- 15.8 ng/ml, p <0.001, respectively) compared with those without embolic events and the control group. Embolic patients also had higher plasminogen activator inhibitor-1 levels than both nonembolic patients and healthy subjects (14.4 +/- 6.4 vs 8.6 +/- 5.9 and 5.4 +/- 4.3 ng/ml, p = 0.002, respectively). In conclusion, IE patients with subsequent thromboembolism have increased systemic coagulation activation, enhanced platelet activity/damage, and impaired fibrinolysis. The resulting imbalance produces a sustained hypercoagulable state that may contribute to the increased risk of thromboembolic events in this particular group. PMID- 12633800 TI - Growth of stenotic lesions after balloon angioplasty for pulmonary artery stenosis after arterial switch operation. AB - Little is known about the growth potential of pulmonary stenotic lesions after balloon angioplasty (BA) in patients after the arterial switch operation. The aim of this study was to evaluate the growth potential of pulmonary stenotic lesions after BA and assess the midterm results of BA for pulmonary artery stenosis after the arterial switch operation. Thirty-seven patients who had undergone 52 procedures had repeat catheterization at a median of 43 years (range 1.2 to 9.3 ys) after BA. To adjust growth-related changes in the size of the pulmonary artery, the stenotic diameter was expressed as a percentage of normal (%N). An immediate increase of 63 +/- 45% in the stenotic diameter and a reduction of 51 +/- 33% in the pressure gradient occurred across the stenotic lesions after BA. The right ventricular-aortic systolic pressure ratio decreased from 0.67 +/- 0.24 to 0.51 +/- 0.12 after BA (p <0.0001). Compared with immediate data after BA, there was no significant change in the growth-adjusted diameter of the stenotic lesions (68 +/- 26 %N after BA vs 65 +/- 25 %N at follow-up, p = 0.08), and the pressure gradient (16 +/- 13 mm Hg after BA vs 20 +/- 21 mm Hg at follow-up, p = 0.10). The ventricular-aortic systolic pressure ratio also did not change (0.51 +/- 0.12 after BA vs 0.50 +/- 0.21 at follow-up, p = 0.57). Restenosis occurred in 3 of 26 vessels (12%) after successful BA in which the diameter increased >50% after BA. Our data suggest that pulmonary stenotic lesions after BA develop with age in growing children after the arterial switch operation, and the efficacy of the BA may be long lasting. PMID- 12633801 TI - Unusual left ventricular dilatation without functional or biochemical impairment in normotensive extremely overweight Japanese professional sumo wrestlers. AB - To explore the physiologic limit of left ventricular (LV) enlargement, we performed echocardiography and air displacement plethysmography to respectively assess LV dimension and function and the body composition of Japanese professional sumo wrestlers. After excluding subjects with cardiovascular disease, hypertension, plasma brain natriuretic peptide (BNP) > or =17.9 pg/ml, diabetes mellitus, or asthma, 331 subjects (mean +/- SD age, 21.6 +/- 3.7 years; height 179.2 +/- 5.3 cm; weight 1,17.9 +/- 21.5 kg; percent fat, 29.6 +/- 6.6%) were analyzed. LV end-diastolic dimension averaged 58.4 +/- 3.7 mm and was within the generally regarded normal limit (< or =54 mm) in 14.5% of subjects, but was > or =60 mm in 41.1% of subjects. LV septal and posterior wall thicknesses were 10.3 +/- 0.9 and 10.2 +/- 0.9 mm, respectively. Peak E- and A-wave velocities, E/A ratio, LV fractional shortening, and BNP were 96 +/- 16 and 51 +/- 13 cm/s, 2.0 +/- 0.7, 33.5 +/- 4.5%, and 3.1 +/- 3.7 pg/ml, respectively. LV end-diastolic dimension was not correlated with these indexes of LV function or with plasma BNP levels, but was significantly correlated with height, weight, body surface area, fat-free mass, and fat mass. These results show that among very large, highly trained, professional athletes, LV end-diastolic dimension frequently exceeds the traditionally accepted upper limit of normal for the general population. This increase in LV end-diastolic dimension may thus represent an extreme example of the physiologic adaptation of the athlete's heart. PMID- 12633802 TI - Outcomes of clinically significant idiopathic pericardial effusion requiring intervention. AB - There is a paucity of outcome data on patients with idiopathic pericardial effusion requiring intervention. All patients who had clinically significant pericardial effusion confirmed by echocardiography and requiring interventions between 1979 and 2000 were identified through the Echo-guided Pericardiocentesis Registry and Echocardiography and Surgical Databases. Clinical data and outcomes were obtained by review of medical records and surveys. The study population consisted of 92 patients (mean age 59 +/- 15 years). Five patients were referred directly for pericardiectomy (3 had effusion in the context of chronic relapsing pericarditis, 2 had effusive constrictive disease), and 87 underwent echo-guided pericardiocentesis as their initial treatment. In 47 of these patients, primary management involved extended pericardial catheter drainage, which was associated with a trend to lower recurrence rates than in those without catheter drainage (p = 0.052). Three patients had transient right ventricular entry with no sequelae, and 7 patients (8%) later had surgical pericardiectomy because of the recurrence of effusion, 2 of whom were also found to have evidence of effusive constrictive disease during surgery. One patient had bleeding after pericardiectomy that required repeat thoracotomy. Mean follow-up of the cohort was 3.8 +/- 4.3 years. For most patients with clinically significant idiopathic pericardial effusion requiring intervention, echo-guided pericardiocentesis was the definitive treatment. Pericardiectomy was necessary for patients in whom effusion occurred in the context of effusive constrictive disease, chronic relapsing pericarditis, or recurrent effusion despite pericardiocentesis. The prognosis for the cohort was favorable, and survival did not appear to differ from that of the general population (p = 0.372). PMID- 12633803 TI - Frequency and sequelae of ST elevation acute myocardial infarction caused by spontaneous distal embolization from unstable coronary lesions. PMID- 12633804 TI - Heart rate recovery after exercise as a predictor of mortality among survivors of acute myocardial infarction. PMID- 12633805 TI - Composition of coronary atherosclerotic plaques in patients with acute myocardial infarction and stable angina pectoris determined by contrast-enhanced multislice computed tomography. PMID- 12633806 TI - Relation between pet ownership and heart rate variability in patients with healed myocardial infarcts. PMID- 12633807 TI - Relation of insurance status to performance of coronary angiography in patients with unstable angina pectoris or acute myocardial infarction. PMID- 12633808 TI - Left ventricular hypertrophy and the risk of subsequent left ventricular remodeling following myocardial infarction. PMID- 12633809 TI - Relation of total annual coronary angioplasty volume of physicians and hospitals on outcomes of primary angioplasty for acute myocardial infarction (data from the 1995 Coronary Angioplasty Reporting System of the New York State Department of Health). PMID- 12633810 TI - Relation of admission white blood cell count to long-term outcomes after primary coronary angioplasty for acute myocardial infarction (The Stent PAMI Trial). PMID- 12633811 TI - Effect of short-term antibiotic treatment on Chlamydia pneumoniae and peripheral endothelial function. PMID- 12633812 TI - Saphenous vein graft stenting using a novel filter device for distal protection. PMID- 12633813 TI - Serial volumetric intravascular ultrasound assessment of native coronary artery versus saphenous vein grafts in-stent restenosis lesions after conventional catheter-based treatment. PMID- 12633814 TI - Safe and efficacious use of bivalirudin for percutaneous coronary intervention with adjunctive platelet glycoprotein IIb/IIIa receptor inhibition. PMID- 12633815 TI - Predictors of aortic and coronary artery calcium on a screening electron beam tomographic scan. PMID- 12633816 TI - Adrenal/renal contribution to circulating norepinephrine in posturally induced neurally mediated reflex syncope. PMID- 12633818 TI - The R-on-T phenomenon in patients with implantable cardioverter-defibrillators. PMID- 12633817 TI - Effects of calcium treatment on QT interval and QT dispersion in hypocalcemia. PMID- 12633819 TI - Usefulness of brain natriuretic peptide release as a surrogate marker of the efficacy of long-term cardiac resynchronization therapy in patients with heart failure. PMID- 12633820 TI - Effect of lower body negative pressure in patients with mild congestive heart failure. PMID- 12633821 TI - Anthropometric normalization of left ventricular size in chronic mitral regurgitation. PMID- 12633822 TI - Location, size, and morphologic characteristics of left atrial thrombi as assessed by transesophageal echocardiography in relation to systemic embolism in patients with rheumatic mitral valve disease. PMID- 12633823 TI - Effects of deflazacort on left ventricular function in patients with Duchenne muscular dystrophy. PMID- 12633824 TI - Relation of pulmonary venous wedge pressures to pulmonary artery pressures in patients with single ventricle physiology. PMID- 12633825 TI - Electrocardiographic changes in thyrotoxic periodic paralysis. PMID- 12633826 TI - Natural history of intramural hematoma of the descending thoracic aorta. PMID- 12633827 TI - Impact of left ventricular ejection fraction on estimation of left ventricular filling pressures using tissue Doppler and flow propagation velocity. PMID- 12633829 TI - 35Cl quadrupolar constants obtained by solid-state NMR: study of chlorinated Al-O P clusters, involving OH...Cl hydrogen bonds. AB - Simulation of static and MAS NMR spectra obtained at high field (B(0) = 14.08 T) and high spinning frequency (up to nu(rot) = 22 kHz) allows the determination of 35Cl quadrupolar parameters for various cubane-shaped, cage-like and cyclic Al-O P clusters; the role of OH...Cl contacts is emphasized. PMID- 12633828 TI - Does phase cycling work for nuclei experiencing strong quadrupolar couplings? AB - The question of whether the phase cycling can still be used to select coherence transfer pathways in spin systems experiencing a tilting of its eigenstates away from the Zeeman eigenstates due to strong couplings was investigated theoretically. Based on the analysis presented it is concluded that conventional phase cycling is still a valid approach for selecting a coherence transfer pathway signal, although changes in pathway efficiencies can occur as the mechanisms for excitation and detection of coherences are affected by the tilting of the eigenstates. PMID- 12633830 TI - An analytical formulation of CPMAS. AB - An exact expression for the cross polarization between two spin-1/2 particles is derived from the quantum Liouville equation. This is given in the form of two integrodifferential equations. These can be solved exactly in the static case (no sample spinning) and a powder average easily performed numerically. With magic angle spinning, the neglect of certain interference terms simplifies the numerical calculation. A further assumption decoupling the calculation of the sidebands gives a very simple formula that is capable of giving a qualitative interpretation of all experimental observations. Examples are given illustrating typical buildup curves and CPMAS matching profiles. PMID- 12633831 TI - X-[1H, 19F] triple resonance with a X-[1H] CP MAS probe and characterisation of a 29Si-19F spin pair. AB - An economic approach for implementing X-[1H,19F] double-decoupling MAS NMR experiments with a conventional X-[1H] dual-channel CP MAS probe is demonstrated. The parameters characterising the isolated 29Si-19F spin pair in an organosilicon compound R(3)SiF (R = 9-anthryl) are determined. In addition, we discuss the optimum choice of experimental parameters for determining all 29Si-19F spin-pair parameters from straightforward 29Si MAS NMR spectra with only 1H decoupling applied during acquisition. PMID- 12633832 TI - A combined 14N/27Al nuclear magnetic resonance and powder X-ray diffraction study of impurity phases in beta-sialon ceramics. AB - Beta-sialons are ceramic phases occurring in the SiO(2)-Si(3)N(4)-AlN-Al(2)O(3) system. A series of samples with differing compositions has been investigated by magic-angle spinning nuclear magnetic resonance (NMR) spectroscopy and powder X ray diffraction (XRD). Although the constituent nitrogen nuclei occupy positions of low symmetry in the beta-sialon structure, 14N NMR spectra could be recorded for the samples examined. The origin of the 14N signal could be traced to the presence of an aluminium nitride (AlN) impurity phase with the help of 27Al NMR and XRD results. Similarly, the existence of Al(2)O(3) grains could be readily detected for a number of samples. Thus, the combination of 14N and 27Al NMR is shown to be an especially effective tool in identifying and characterizing impurity phases in sialon ceramics, complementing the results obtained from standard XRD analysis. PMID- 12633833 TI - 13C CP MAS NMR and GIAO-CHF calculations of coumarins. AB - 13C cross-polarization magic-angle spinning NMR spectra were recorded for a series of solid coumarins. Ab initio calculations of shielding constants were performed with the use of GIAO-CHF method. The combined CPMAS NMR and theoretical approach was successful in characterizing solid-state conformations of coumarins; a relationship sigma (ppm) = -1.032 xdelta + 205.28 (R(2) = 0.9845) can be used to obtain structural information for coumarins, for which solid-state NMR or crystal structure data are not available. PMID- 12633834 TI - The effects of different heat treatment and atmospheres on the NMR signal and structure of TiO2-ZrO2-SiO2 sol-gel materials. AB - The effects of different heat treatment schemes (i.e. successively or directly heated to particular temperatures) and atmospheres (air or nitrogen) on the solid state NMR spectra obtained from (TiO(2))(0.15)(ZrO(2))(0.05)(SiO(2))(0.80) sol gel materials are investigated. A combination of 1H, 13C, 17O and 29Si NMR is used. 29Si MAS NMR indicates that the extent of condensation of the silica-based network strongly depends on the maximum temperature the sample has experienced, but the condensation is largely independent of the details of the heat treatment scheme and atmosphere used. For sol-gel produced silicate-based materials the results show that the equilibrium structure at each temperature is reached rapidly compared to the time (2h) spent at that temperature. The 17O NMR results confirm that a nitrogen atmosphere does significantly reduce loss of 17O from the structure but care must be taken since there could be differential loss of 17O from the regions having different local structural characteristics. PMID- 12633835 TI - Small 51V chemical shift anisotropy for LaVO4 from MQMAS and MAS NMR spectroscopy. AB - 51V MQMAS NMR of the triple-quantum transitions is shown to be particularly useful in the determination of the sign and magnitude of the chemical shift anisotropy (CSA) parameter delta(sigma)(= delta(iso)-delta(zz)) along with the asymmetry parameter (eta(sigma)) for a vanadium environment with a small CSA and a rather strong quadrupole coupling. This is demonstrated for the orthovanadate LaVO(4) for which 51V magic-angle spinning (MAS) NMR of the central and satellite transitions at 14.1T gives precise values for the quadrupole coupling parameters, however, an ambiguous sign for delta(sigma). The CSA parameters are reliably obtained from analysis of the spinning sidebands observed in a 51V triple-quantum MAS experiment. Combining these data with least-squares analysis of the manifold of spinning sidebands in the single-pulse MAS NMR spectrum results in a precise determination of the magnitudes and relative orientation of the 51V quadrupole coupling and CSA tensors for LaVO(4). PMID- 12633836 TI - 95Mo NMR in Mo3 Se4 [Addendum to Solid State Nucl. Magn. Reson. 12 (1998) 191]. PMID- 12633837 TI - Angiogenesis in hematologic malignancies. AB - Angiogenesis is defined as the formation of new capillaries from preexisting blood vessels and plays an important role in the progression of solid tumors. Recently a similar relationship has been described in several hematologic malignancies. Expression of the angiogenic peptides vascular endothelial growth factor (VEGF) and basic fibroblast growth factor correlates with clinical characteristics in leukemia and non-Hodgkin's-lymphoma and the serum/plasma concentrations serve as predictors of poor prognosis. Increased bone marrow microvessels in multiple myeloma (MM) are correlated with decreased overall survival. Thalidomide which has antiangiogenic effects and direct cytotoxic effects was found to be effective in MM, myelodysplastic syndrome and acute myeloid leukemia (AML). Preliminary data indicate activity of VEGF-tyrosine kinase inhibitors in AML. Clinical research is now aimed at testing antiangiogenic treatment strategies in several hematologic neoplasms as well as identifying the best candidate patients for specific approaches. PMID- 12633838 TI - Malignant melanoma: current state of primary and adjuvant treatment. AB - Metastatic malignant melanoma remains a highly lethal disease with an incidence that continues to rise. Management of melanoma includes definitive local, regional and distant control. There is substantial prospective and retrospective data to base the extent of both primary as well as adjuvant therapy. The results of these trials have on occasion been at odds. A critical assessment of the available information pertaining to the adjuvant treatment of cutaneous melanoma is needed. This review provides a critical assessment of the current data that is available to guide both primary resection as well as adjuvant therapy. To date, current trials have shown little promise with nonspecific immunostimulants and cytotoxic chemotherapy. In contrast, dose interferon-alpha2b has been shown to improve relapse-free survival and likely improves melanoma-specific survival as well. Based on the available data, interferon-alpha2b remains the adjuvant therapy of choice for high-risk patients treated outside clinical trials, and the appropriate control arm for clinical trials evaluating new or modified adjuvant regimens. PMID- 12633839 TI - An overview of current results with the gemcitabine and docetaxel combination as initial and salvage chemotherapy regimen in advanced non-small cell lung cancer. AB - The activity of single-agent docetaxel in patients with platinum pretreated non small cell lung cancer (NSCLC) has been established in two recent large randomized phase III trials, either against best supportive care or versus single agent vinorelbine/ifosfamide. Moreover, single-agent gemcitabine has demonstrated significant activity and clinical benefit in platinum- and paclitaxel-pretreated advanced NSCLC. Combination regimens employing these two agents in various doses and schedules have recently been initiated. The gemcitabine/docetaxel combination with or without G-CSF support as salvage therapy of NSCLC pre-treated with platinum+/-paclitaxel-based regimens has been evaluated in four recently published phase II clinical studies and has been shown to represent a tolerable and active regimen in this setting, yielding a 10-33% response rate, thus, warranting randomized comparisons to single-agent gemcitabine or docetaxel, drugs currently recommended in second-line treatment of advanced NSCLC. PMID- 12633840 TI - The relationship between aging and carcinogenesis: a critical appraisal. AB - The incidence of cancer increases with age in humans and in laboratory animals alike. There are different patterns of age-related distribution of tumors in different organs and tissues. Aging may increase or decrease the susceptibility of various tissues to initiation of carcinogenesis and usually facilitates promotion and progression of carcinogenesis. Aging may predispose to cancer by several mechanisms: (1) tissue accumulation of cells in late stages of carcinogenesis; (2) alterations in homeostasis, in particular, alterations in immune and endocrine system and (3) telomere instability linking aging and increased cancer risk. Increased susceptibility to the effects of tumor promoters is found both in aged animals and aged humans, as predicted by the multistage model of carcinogenesis. Available evidence supporting the relevance of replicative senescence of human cells and telomere biology to human cancer seems quite strong, however, the evidence linking cellular senescence to human aging is controversial and required additional studies. Data on the acceleration of aging by carcinogenic agents as well as on increased cancer risk in patients with premature aging are critically discussed. In genetically modified mouse models (transgenic, knockout or mutant) characterized by the aging delay, the incidence of tumors usually similar to those in controls, whereas the latent period of tumor development is increased. Practically all models of accelerated of aging in genetically modified animals show the increase in the incidence and the reduction in the latency of tumors. Strategies for cancer prevention must include not only measures to minimize exposure to exogenous carcinogenic agents, but also measures to normalize the age-related alterations in internal milieu. Life-span prolonging drugs (geroprotectors) may either postpone population aging and increase of tumor latency or decrease the mortality in long-living individuals in populations and inhibit carcinogenesis. At least some geroprotectors may increase the survival of a short-living individuals in populations but increase the incidence of malignancy. PMID- 12633841 TI - Short course radiation therapy for elderly cancer patients. Evidences from the literature review. AB - The choice of the appropriate treatment strategy for elderly cancer patients may be a difficult challenge. Radiation therapy is commonly offered to these patients, but treatment duration may represent a limiting factor, as many patients cannot tolerate a conventional course of radiotherapy (RT) due to age related medical or logistic problems. Hypofractionated RT may represent a very convenient choice, but it entails an increased risk of late toxicity occurrence. We made a literature review to define the possible role of hypofractionated RT for elderly cancer patients. As expected, we found out that short irradiation schedules are more commonly employed for treatments with palliative aims but a more widespread use of these regimes is still controversial. The lack of prospective trials tailored for these patients makes even more difficult to tailor the choice of treatment on standardised treatment guidelines. Nevertheless our review highlights that for several tumour types RT can be scheduled conveniently and effectively in order to achieve local disease control and/or symptom relief with the least discomfort and treatment-related morbidity for elderly patients. PMID- 12633842 TI - Primary breast cancer in elderly women: biological profile and relation with clinical outcome. AB - A biological characterisation carried out on 14,007 primary breast cancers provided evidence in favour of a relation between advanced age and favourable features (positivity for oestrogen/progesterone receptors, low proliferative rate, absence of p53 accumulation, bcl-2 overexpression, diploid DNA content), showed a similar pattern of association between patho-biological variables regardless of patient age, and demonstrated a relation between biological variables and disease outcome in the elderly, comparable to that already reported for younger patients. In fact, oestrogen receptor and proliferative activity provided independent prognostic information either in node-negative or in node positive tumours treated with radical or conservative surgery plus radiotherapy, alone or followed by adjuvant hormonal therapy. It would be thus reasonable to use biomarkers as a complement to clinico-pathological features in a 'risk-factor profile system' even for elderly patients, upon their validation in prospective studies and after assessing the cost-benefit of treatments planned on the basis of biological information. PMID- 12633843 TI - A hydrophobic platform as a mechanistically relevant transition state stabilising factor appears to be present in the active centre of all glycoside hydrolases. AB - An in silico survey of the -1 subsite of all known 3D-structures of O-glycoside hydrolases containing a suitably positioned ligand has led to the recognition -- apparently without exceptions -- of a transition state stabilising hydrophobic platform which is complementary to a crucial hydrophobic patch of the ligand. This platform is family-specific and highly conserved. A comprehensive list is given with examples of enzymes belonging to 33 different families. Several typical constellations of platform - protein residues are described. PMID- 12633844 TI - Structure-function relationships of the extracellular domain of the autosomal dominant polycystic kidney disease-associated protein, polycystin-1. AB - Polycystin-1 (PC-1) is a member of a novel family of proteins that have a multidomain structure. Although the C-terminal intracellular segments have been extensively studied, mainly with respect to their putative involvement in cell signalling, the potential function of the extracellular domains has received less attention. Mutations in PC-1 result in autosomal dominant polycystic kidney disease (ADPKD) which is characterised by perturbation of transport resulting in fluid accumulation, cell proliferation and modification of the extracellular matrix. The possibility that the interaction of a component of the extracellular matrix or some external factor with PC-1 may be important in the initiation or progression of ADPKD cannot currently be ruled out. The purpose of this review is to assess current evidence for the function of the PC-1 extracellular domains, and their potential implications for ADPKD. PMID- 12633845 TI - Functional role of sperm surface glutathione S-transferases and extracellular glutathione in the haploid spermatozoa under oxidative stress. AB - On the sperm surface, glutathione S-transferases (GSTs) exist as oocyte binding proteins but their detoxification function in this unique cell type is not known. Using H(2)O(2)- and 4-hydroxynonenal-induced sperm dysfunction models, this study demonstrates that the sperm surface GSTs are able to use extracellular reduced glutathione to inhibit the loss of functional competence of goat spermatozoa; however, in the presence of GST inhibitors, they are unable to do so. In the context of susceptibility of spermatozoa to oxidative stress, this finding that strategically located sperm surface GSTs are important for maintaining the functional competence of sperm is relevant to studies on male infertility. PMID- 12633846 TI - Human colon cancer cells differ in their sensitivity to curcumin-induced apoptosis and heat shock protects them by inhibiting the release of apoptosis inducing factor and caspases. AB - Mild heat treatment induced the expression of heat shock protein-70 (hsp70), hsp90 and hsp27 in two human colon cancer cell lines, one derived from primary tumor, SW480, and the other derived from the secondary lymph node tissue, SW620, of the same patient. SW620 cells appear to be more sensitive to curcumin-induced apoptosis. Heat shock protects both the human colon cancer cells from curcumin induced apoptosis. Heat shock prevented, at least in part, the release of apoptosis inducing factor from mitochondria induced by curcumin although the release of second mitochondria derived activator of caspase and cytochrome c was unaffected in both the cells. Moreover, heat shock reduced curcumin-induced activation of caspases 9 and 3 but not 8. PMID- 12633847 TI - Novel two-color fluorescence probe with extreme specificity to bovine serum albumin. AB - We report on strong, highly specific and stochiometric binding to bovine serum albumin of novel fluorescence probe FA, 2-(6-diethylaminobenzo[b]furan-2-yl)-3 hydroxychromone, that exhibits a very characteristic two-band fluorescence spectrum. Both absorption band and two fluorescence bands of FA are very sensitive to non-covalent interactions in the immediate environment of the probe. Multiparametric analysis of this spectroscopic information allows us to conclude that the binding site is characterized by very low polarity, high extent of screening from aqueous environment and unusually high electronic polarizability. The latter suggests the proximal location of probe FA to the aromatic amino acid residues in the binding site. The new probe can be proposed for the study of interaction of ligands and drugs of different nature with serum albumins. PMID- 12633848 TI - A cadherin-like protein functions as a receptor for Bacillus thuringiensis Cry1Aa and Cry1Ac toxins on midgut epithelial cells of Bombyx mori larvae. AB - Aminopeptidase N (APN) and cadherin-like protein (BtR175) from Bombyx mori larvae were examined for their roles in Cry1Aa- and Cry1Ac-induced lysis of B. mori midgut epithelial cells (MECs). APNs and BtR175 were present in all areas of the midgut, were particularly abundant in the posterior region, and were found only on columnar cell microvilli and not on the lateral membrane that makes cell-cell contacts. This distribution was in accordance with the distribution of Cry1A susceptible MECs in the midgut. The lytic activity of Cry1Aa and Cry1Ac on collagenase-dissociated MECs was linearly dependent on toxin concentration. Although pre-treatment of MECs with anti-BtR175 antibody was observed to partially inhibit the lytic activity exerted by 0.1-1 nM Cry1Aa toxin or 5 nM Cry1Ac toxin, no significant inhibition was observed when MECs were pre-treated with anti-APN antibody. These results suggest that BtR175 functions as a major receptor for Cry1A toxins in the midgut of B. mori larvae. PMID- 12633849 TI - Purification of active recombinant trypanosome alternative oxidase. AB - Trypanosome alternative oxidase (TAO) is the terminal oxidase of the respiratory chain in long slender bloodstream forms of African trypanosomes. TAO is a cytochrome-independent, cyanide-insensitive quinol oxidase. These characteristics are distinct from those of the bacterial quinol oxidases, proteins that belong to the heme-copper terminal oxidase superfamily. The inability to purify stable TAO has severely hampered biochemical studies of the alternative oxidase family. In the present study, we were able to purify recombinant TAO to homogeneity from Escherichia coli membranes using the detergent digitonin. Kinetic analysis of the purified TAO revealed that the specific inhibitor ascofuranone is a competitive inhibitor of ubiquinol oxidase activity. PMID- 12633850 TI - Identification of a novel Bcl-xL phosphorylation site regulating the sensitivity of taxol- or 2-methoxyestradiol-induced apoptosis. AB - Bcl-xL, a close homolog of Bcl2, is an important regulator of apoptosis and is overexpressed in human cancer. Phosphorylation of Bcl-xL can be induced by microtubule-damaging drugs such as taxol or 2-methoxyestradiol (2-ME). By site directed mutagenesis studies, we have identified that serine 62 is the necessary site for taxol- or 2-ME-induced Bcl-xL phosphorylation in prostate cancer cells. Further studies with the inhibitor of Jun kinase (JNK) and phosphorylation null mutant of Bcl-xL reveal the augmentative role of JNK-mediated Bcl-xL phosphorylation in apoptosis of prostate cancer cells. In summary, our studies suggest that the phosphorylation of Bcl-xL by stress response kinase signaling might oppose the anti-apoptotic function of Bcl-xL to permit prostate cancer cells to die by apoptosis. PMID- 12633851 TI - Specific conjugation of DNA binding proteins to DNA templates through thiol disulfide exchange. AB - The double-stranded oligodeoxyribonucleotides with single internucleotide disulfide linkages were successfully used for covalent trapping of cysteine containing protein. In particular, an efficient conjugation of DNA methyltransferase SsoII to sequence-specific decoys was demonstrated. The obtained results assume that synthetic oligodeoxyribonucleotides bearing a new trapping site can be used as new tools to study and manipulate biological systems. PMID- 12633852 TI - Fusion of mitochondria in mammalian cells is dependent on the mitochondrial inner membrane potential and independent of microtubules or actin. AB - Mitochondrial fusion is a poorly characterized process which has mainly been studied in yeast and Drosophila but is thought to occur in all eukaryotes. Until now, there was only indirect evidence to support such a process in mammalian cells. In this study, using a cell fusion system, we found that mitochondrial fusion occurs rapidly in mammalian cells and is completed in less than 24 h. We report that the fusion of mitochondria requires an intact mitochondrial inner membrane potential but is independent of a functional cytoskeleton. PMID- 12633853 TI - The hydrolysis of lysophospholipids and nucleotides by autotaxin (NPP2) involves a single catalytic site. AB - Autotaxin (NPP2) is a tumor cell motility-stimulating factor that displays both a nucleotide pyrophosphatase/phosphodiesterase activity and a recently described lysophospholipase D activity. The hydrolysis of nucleotides is a metal-assisted reaction that occurs via a nucleotidylated threonine in the catalytic site. We show here that the catalytic site threonine and the metal-coordinating residues are also essential for the hydrolysis of lysophospholipids. In comparing the substrate specificity of NPP2 and the closely related NPP1 and NPP3, we found that only NPP2 displayed a lysophospholipase D activity, whereas NPP1 and NPP3 had a much higher nucleotide pyrophosphatase activity. PMID- 12633854 TI - Crystal structure of Drosophila angiotensin I-converting enzyme bound to captopril and lisinopril. AB - Angiotensin I-converting enzymes (ACEs) are zinc metallopeptidases that cleave carboxy-terminal dipeptides from short peptide hormones. We have determined the crystal structures of AnCE, a Drosophila homolog of ACE, with and without bound inhibitors to 2.4 A resolution. AnCE contains a large internal channel encompassing the entire protein molecule. This substrate-binding channel is composed of two chambers, reminiscent of a peanut shell. The inhibitor and zinc binding sites are located in the narrow bottleneck connecting the two chambers. The substrate and inhibitor specificity of AnCE appears to be determined by extensive hydrogen-bonding networks and ionic interactions in the active site channel. The catalytically important zinc ion is coordinated by the conserved Glu395 and histidine residues from a HExxH motif. PMID- 12633855 TI - ATP-induced, sustained calcium signalling in cultured rat cortical astrocytes: evidence for a non-capacitative, P2X7-like-mediated calcium entry. AB - The receptor mechanisms regulating the ATP-induced free cytosolic Ca(2+) concentration ([Ca(2+)](i)) changes in cultured rat cortical type-1 astrocytes were analyzed using fura-2-based Ca(2+) imaging microscopy. Upon prolonged ATP challenge (1-100 microM), astroglial cells displayed a biphasic [Ca(2+)](i) response consisting of an initial peak followed by a sustained elevation. Suramin and pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid blocked both components, albeit to a different extent. By contrast, the selective P2X7 antagonist oxidized ATP irreversibly abrogated the sustained [Ca(2+)](i) signal without affecting the transient phase. Finally, astrocyte challenge with the selective P2X7 agonist 3' O-(4-benzoyl)benzoyl-ATP evoked a sustained [Ca(2+)](i) elevation, which occluded that induced by ATP. We can conclude that in cultured cortical astrocytes the ATP mediated sustained [Ca(2+)](i) rise does not implicate capacitative Ca(2+) entry but involves Ca(2+) influx through P2X7-like receptors. PMID- 12633856 TI - Intramolecular rearrangements as a consequence of the dephosphorylation of phosphoaspartate residues in proteins. AB - Aspartate phosphorylation induces changes in protein conformation that are used to regulate processes ranging from gene expression and cell differentiation to cell motility and the generation of electrochemical gradients across membranes. We show here that dephosphorylation of the phosphoaspartate in the chemotaxis response regulator CheY can result in the loss of a water molecule that may be due to formation of a succinimide intermediate. PMID- 12633857 TI - Coenzyme M binds to a [4Fe-4S] cluster in the active site of heterodisulfide reductase as deduced from EPR studies with the [33S]coenzyme M-treated enzyme. AB - Heterodisulfide reductase (Hdr) from methanogenic Archaea catalyzes the reversible reduction of the heterodisulfide (CoM-S-S-CoB) of the methanogenic thiol coenzymes, coenzyme M (CoM-SH) and coenzyme B (CoB-SH). Upon reaction of the oxidized enzyme with CoM-SH a unique paramagnetic species is formed, which has been shown to be due to a novel type of [4Fe-4S](3+) cluster. In this work, it was addressed whether CoM-SH is directly attached to this [4Fe-4S] cluster using CoM-(33)SH as substrate and purified Hdr from Methanothermobacter marburgensis and Methanosarcina barkeri. With both enzymes treatment with CoM (33)SH in the presence of duroquinone as an oxidant resulted in a significant broadening of the electron paramagnetic resonance spectrum as compared to CoM-SH as substrate. The signal broadening resulted from an unresolved anisotropic hyperfine coupling between the (33)S nucleus and the paramagnetic center. The results provide compelling evidence for a direct binding of CoM-SH to the [4Fe 4S] cluster in the active site of the enzyme. PMID- 12633858 TI - Involvement of caveolin-2 in caveolar biogenesis in MDCK cells. AB - Caveolins have been identified as key components of caveolae, specialized cholesterol-enriched raft domains visible as small flask-shaped invaginations of the plasma membrane. In polarized MDCK cells caveolin-1 and -2 are found together on basolateral caveolae whereas the apical membrane, where only caveolin-1 is present, lacks caveolae. Expression of a caveolin mutant prevented the formation of the large caveolin-1/-2 hetero-oligomeric complexes, and led to intracellular retention of caveolin-2 and disappearance of caveolae from the basolateral membrane. Correspondingly, in MDCK cells over-expressing caveolin-2 the basolateral membrane exhibited an increased number of caveolae. These results indicate the involvement of caveolin-2 in caveolar biogenesis. PMID- 12633859 TI - The active site of pepsin is formed in the intermediate conformation dominant at mildly acidic pH. AB - Pepsin is an aspartic protease that acts in food digestion in the mammal stomach. An optimal pH of around 2 allows pepsin to operate in its natural acidic environment, while at neutral pH the protein is denatured. Although the pH dependence of pepsin activity has been widely investigated since the 40s, a renewed interest in this protein has been fueled by its homology to the HIV and other aspartic proteases. Recently, an inactive pepsin conformation has been identified that accumulates at mildly acidic pH, whose structure and properties are largely unknown. In this paper, we analyse the conformation of pepsin at different pHs by a combination of spectroscopic techniques, and obtain a detailed characterisation of the intermediate. Our analysis indicates that it is the dominant conformation from pH 4 to 6.5. Interestingly, its near UV circular dichroism spectrum is identical to that of the native conformation that appears at lower pH values. In addition, we show that the intermediate binds the active site inhibitor pepstatin with a strength similar to that of the native conformation. Pepsin thus adopts, in the 6.5-4.0 pH interval, a native-like although catalytically inactive conformation. The possible role of this intermediate during pepsin transportation to the stomach lumen is discussed. PMID- 12633860 TI - Identification of cDNAs from Japanese pufferfish (Fugu rubripes) and Atlantic salmon (Salmo salar) coding for homologues to tetrapod prion proteins. AB - We identified cDNAs coding for homologues to tetrapod prion proteins (PrPs) in Atlantic salmon (Salmo salar) and Japanese pufferfish (Fugu rubripes), which were termed 'similar to PrPs' (stPrPs). Besides significant sequence homologies the fish stPrPs display characteristic structural features in common with tetrapod PrPs. In addition, two stPrPs were shown to be highly expressed in brain tissue. None of the so far identified PrP-homologues of fish resembles doppel. Hence, the duplication of the PrP gene, which generated doppel, may have occurred not in fish but later in the tetrapod lineage. The identification of fish PrPs provides a basis to address concerns about a possible susceptibility of fish to prion infections. PMID- 12633861 TI - Osmomechanical stress selectively regulates translocation of protein kinase C isoforms. AB - Osmomechanical stress, resulting in cell swelling and activation/regulation of numerous cellular processes, may play a critical role in cell signaling by selectively regulating translocation of protein kinase C (PKC) isoforms from cytosol to membrane compartments. Western blotting of renal epithelial cell fractions demonstrated the expression of five PKC isoforms. Three of these isoforms (PKCalpha, PKCepsilon, PKCzeta) translocated to the membrane fraction upon exposure of cells to osmomechanical stress (hypotonic medium). Immunohistochemical staining of cells using isoform-specific antibodies further demonstrated translocation of the phorbol ester-sensitive isoforms, PKCalpha and PKCepsilon, to both the plasma membrane and perinuclear sites, reflecting potential initial steps in regulation of specific effector pathways. Indeed, selective inhibition of PKCs indicates a potential role for PKCalpha in modulating a calcium influx channel. It is concluded that osmomechanical stress induces selective translocation of specific PKC isoforms, demonstrating a key role of osmomechanical stress in selectively regulating PKC-dependent signaling pathways. PMID- 12633862 TI - Biosynthesis of hepoxilins: evidence for the presence of a hepoxilin synthase activity in rat insulinoma cells. AB - The 12(S)-lipoxygenase (12-LOX) pathway of arachidonic acid (AA) metabolism after dioxygenation to 12(S)-hydroperoxy-eicosatetraenoic acid is bifurcated in a reduction route to formation of 12(S)-hydroxy-eicosatetraenoic acid (12-HpETE) and an isomerization route to formation of hepoxilins. Interestingly, we found that the rat insulinoma RINm5F cells, which are devoid of cytoplasmic glutathione peroxidase (cGPx)/phospholipid hydroperoxide glutathione peroxidase (PHGPx), produce solely hepoxilin A(3) (HXA(3)). Since HXA(3) synthesis was abolished in heat-denatured or cGPx- or PHGPx-transfected cells, it was tempting to speculate that a HXA(3) synthase activity regulated by cGPx/PHGPx is present. To confirm this assumption we incubated AA with HeLa cells overexpressing the rat leukocyte type 12-LOX. Neither HXA(3) nor 12(S)-HETE were detected due to abundance of cGPx/PHGPx. But, pretreatment of transfected cells with diethyl maleate, an inhibitor of glutathione and PHGPx, restored HXA(3) synthase and 12-LOX activities. Thus, we conclude, that cells containing rat leukocyte-type 12-LOX also possess an intrinsic HXA(3) synthase activity, which is activated by inhibition of cGPx/PHGPx. In normal cells HXA(3) is down-regulated by cGPx/PHGPx, but, it is persistently activated in oxidatively stressed cells deficient in cGPx/PHGPx, such as RINm5F. PMID- 12633863 TI - The human interleukin-6 (IL-6) receptor exists as a preformed dimer in the plasma membrane. AB - The recently solved X-ray structure of the extracellular portion of the interleukin-6 (IL-6) receptor (IL-6R) revealed an IL-6R dimer in the crystal lattice which probably represents a physiological dimer. Performing coprecipitation experiments with two differently tagged IL-6R variants expressed in COS-7 cells, we show that an IL-6R dimer exists in the plasma membrane in the absence of IL-6. Ligand binding does not seem to affect the dimerization status. When lysates of COS-7 cells expressing only one of the IL-6R variants are mixed, spontaneous dimerization occurs. Thus, the IL-6R dimer observed in the crystal structure represents a physiologically occurring phenomenon. PMID- 12633864 TI - Component plane presentation integrated self-organizing map for microarray data analysis. AB - We describe a powerful approach, component plane presentation integrated self organizing map (SOM), for the analysis of microarray data. This approach allows the display of multi-dimensional SOM outputs of microarray data in multiple sample specific presentations, providing distinct advantages in visual inspection of biological significances of genes clustered in each map unit with respect to each RNA sample. Beneficial potentials of the approach are highlighted by processing microarray data from yeast cells as well as human breast malignancies. PMID- 12633865 TI - In vivo studies of liver-type fatty acid binding protein (L-FABP) gene expression in liver of transgenic zebrafish (Danio rerio). AB - Mammalian liver fatty acid binding protein (L-FABP) is a small cytosolic protein in various tissues including liver, small intestine and kidney and is thought to play a crucial role in intracellular fatty acid trafficking and metabolism. To better understand its tissue-specific regulation during zebrafish hepatogenesis, we isolated 5'-flanking sequences of the zebrafish L-FABP gene and used a green fluorescent protein (GFP) transgenic strategy to generate liver-specific transgenic zebrafish. The 2.8-kb 5'-flanking sequence of zebrafish L-FABP gene was sufficient to direct GFP expression in liver primordia, first observed in 2 dpf embryos and then continuously to the adult stage. This pattern of transgenic expression is consistent with the expression pattern of the endogenous gene. F2 inheritance rates of 42-51% in all the seven transgenic lines were consistent with the ratio of Mendelian segregation. Further, hhex and zXbp-1 morphants displayed a visible liver defect, which suggests that it is possible to establish an in vivo system for screening genes required for liver development. PMID- 12633867 TI - Functional EF-Tu with large C-terminal extensions in an E coli strain with a precise deletion of both chromosomal tuf genes. AB - An Escherichia coli strain was constructed in which both chromosomal genes encoding elongation factor (EF)-Tu (tufA and tufB) have been inactivated with precise coding sequence replacements. A tufA gene in an expression vector is supplied as the sole EF-Tu source. By using plasmid replacement, based on plasmid incompatibility, mutant EF-Tu variants with a large C'-terminal extension up to 270 amino acids were studied and proved to be functional in a strain lacking the chromosomal tufA and tufB genes. PMID- 12633866 TI - Low affinity cholecystokinin receptor inhibits cholecystokinin- and bombesin induced oscillations of cytosolic Ca2+ concentration. AB - We have investigated whether low affinity cholecystokinin (CCK) receptors suppress agonist-induced rises of cytosolic free Ca(2+) concentration ([Ca(2+)]c) in pancreatic acinar cells by using properties of caffeine. A high concentration of caffeine (20 mM) completely blocked inositol 1,4,5-trisphosphate (InsP(3)) induced [Ca(2+)]c rises but spared the InsP(3)-independent long-lasting [Ca(2+)]c oscillations. In the presence of 20 mM caffeine, only high concentrations of CCK, but not bombesin or JMV-180, suppressed the caffeine-resistant CCK or bombesin induced [Ca(2+)]c oscillations, indicating that low affinity CCK receptors inhibit agonist-induced [Ca(2+)]c oscillations. It could be one of the underlying mechanisms by which low affinity CCK receptors suppress secretion in pancreatic acinar cells. PMID- 12633868 TI - Lithium-induced inhibition of Src tyrosine kinase in rat cerebral cortical neurons: a role in neuroprotection against N-methyl-D-aspartate receptor-mediated excitotoxicity. AB - The neuroprotective effects of lithium, a mood stabilizer, against glutamate induced excitotoxicity in rat cortical neurons were associated with a decrease in Tyr1472 phosphorylation of the N-methyl-D-aspartate (NMDA) receptor NR2B subunit and a loss of receptor activity. Since this receptor tyrosine phosphorylation is mediated by the Src-family tyrosine kinases, we investigated the effects of lithium on the Src kinase activity. Levels of phosphorylated Src kinase at Tyr416, an index of Src activation, were reduced after treatment with LiCl (1 mM) for more than 3 days. Protein levels of Src-family kinases such as Src, Fyn, and Yes were unchanged by lithium treatment. The activities of cytosolic protein tyrosine kinase and protein phosphatase were also unchanged by lithium treatment, indicating the selectivity and the modulation. Moreover, the levels of postsynaptic densities (PSD) and SynGAP, the scaffolding proteins of the NMDA receptor complex, were unaltered by lithium. A Src kinase inhibitor, SU6656, and an NR2B antagonist, ifenprodil, partially blocked glutamate excitotoxicity. Our results suggest that lithium-induced inactivation of Src kinase contributes to this drug-induced NMDA receptor inhibition and neuroprotection against excitotoxicity. PMID- 12633869 TI - Nucleoporation of dendritic cells: efficient gene transfer by electroporation into human monocyte-derived dendritic cells. AB - Dendritic cells (DCs) are ideal accessory cells in the developing field of gene therapy. Although viral transfection of DCs has become widespread, non-viral transfection of DCs has shown disappointing results. Recently, a new technique for transfecting primary cells has become available -- the Amaxa Nucleofector. Here, we describe the use of this device in the successful non-viral transfection of human monocyte-derived DCs. Using enhanced green fluorescent protein as a reporter gene DCs were transfectable with efficiencies approaching 60%, remaining responsive to lipopolysaccharide-stimulated cytokine production in short-term experiments (though long-term functional assays were hampered by loss of viability). Although these data demonstrate the ease and efficiency with which human monocyte-derived DCs can now be non-virally transfected, they also suggest the limitations of this technology due to the gradual loss of cell viability. The potential use of this system in the development of DC-based cell and gene therapies will be hampered until cell viability can be maintained. PMID- 12633870 TI - Potato tuber phospholipids contain colneleic acid in the 2-position. AB - Colneleic acid (9-[1'(E),3'(Z)-nonadienyloxy]-8(E)-nonenoic acid) is produced from linoleic acid by the sequential action of 9-lipoxygenase and divinyl ether synthase. We demonstrate that a small fraction of the colneleic acid in potato tubers is esterified in phospholipids. This colneleic acid was released by chemical hydrolysis and a phospholipase A(2), but not by a lipase with 1-acyl specificity. Phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol contain molecular species with nominal masses consistent with identification as palmitoyl,colneleoyl species. Exact mass analysis of its fragments confirmed the identity of palmitoyl,colneloyl phosphatidylinositol. To our knowledge, this work represents the first identification of a colneleoyl phospholipid. PMID- 12633871 TI - Specific detection of non-functional human P2X(7) receptors in HEK293 cells and B lymphocytes. AB - P2X(7) receptor/channels mediate ATP-induced apoptosis in a range of cells including lymphocytes. HEK293 cells were transfected with wild-type human P2X(7) receptor or site-directed mutant constructs (K193A, K311A and E496A) known to be non-functional from measurements of barium/ethidium influx in the presence of ATP or 2',3'-O-(4-benzoylbenzoyl)-ATP. An antibody was designed against an epitope from a loop adjacent to the extracellular ATP site. The epitope was unavailable in cells expressing normal functional surface receptors. Non-functional surface receptors as well as intracellular receptors selectively bound the antibody. So did B-lymphocytes from chronic lymphocytic leukemia patients expressing non functional (E496A) mutant receptor. PMID- 12633872 TI - Developmental regulation of beta-1,3-galactosyltransferase-1 gene expression in mouse brain. AB - beta-1,3-galactosyltransferase-1 (beta3GalT-1) is the key enzyme to form the type 1 chain structure. Northern blot analysis indicated that beta3GalT-1 was expressed predominantly in the brain. In the present study, it was revealed that the gene expression of beta3GalT-1 in mouse brain was developmentally decreased. High expression levels of beta3GalT-1 were found in cerebral cortex and hippocampus in both newborn and adult mice, while in cerebellum, the expression levels decreased markedly during development. In situ hybridization revealed that the absence of expression in cerebellar granual cell layers contributed to the main loss of beta3GalT-1 expression in adult mouse cerebellum. Moreover, the decreased levels of beta3GalT-1 could affect the synthesis of type 1 chain oligosaccharides, as revealed by immunohistochemistry analysis. PMID- 12633873 TI - Ets, Ap-1 and GATA factor families regulate the utrophin B promoter: potential regulatory mechanisms for endothelial-specific expression. AB - Duchenne muscular dystrophy is caused by dystrophin deficiency, which can be prevented in the mdx mouse model by over-expression of an autosomal homologue, utrophin. Utrophin has two characterised full-length promoters, A and B. No data are available on the transcriptional regulation of B utrophin, which has been recently localised to the endothelium. Similar to characterised endothelial promoters, Ets and Ap-1 individually trans-activate the human B core promoter. Synergistic activation by GATA-2 and c-jun to the order of 20-fold was observed. PMID- 12633874 TI - Identification of mammalian orthologs associates PYPAF5 with distinct functional roles. AB - PYRIN- and CARD-containing proteins belong to a recently identified protein family involved in the regulation of apoptosis and inflammatory processes. Variations in the gene products of the family members PYPAF1 and NOD2/CARD15 have been associated with several autoinflammatory diseases. We could identify the mouse orthologs of PYPAF1, PYPAF5, NOD1, NOD2 and the rat ortholog of PYPAF5. Intriguingly, we found that PYPAF5 has been reported previously not only as regulator of NF-kappaB and caspase-1, but also as angiotensin II and vasopressin receptor. In particular, based on a comprehensive sequence analysis, we propose a structural model for this hormone receptor that is different from the model suggested previously. PMID- 12633875 TI - Ribosome-inactivating proteins depurinate poly(ADP-ribosyl)ated poly(ADP-ribose) polymerase and have transforming activity for 3T3 fibroblasts. AB - It has been known that ribosome-inactivating proteins (RIPs) from plants damage ribosomes by removing adenine from a precise position of rRNA. Subsequently it was observed that all tested RIPs depurinate DNA, and some of them also non ribosomal RNAs and poly(A), hence the denomination of adenine polynucleotide glycosylases was proposed. We report now that ricin, saporin-L2, saporin-S6, gelonin and momordin depurinate also poly(ADP-ribosyl)ated poly(ADP-ribose) polymerase (auto modified enzyme), an enzyme involved in DNA repair. We observed also that all RIPs but gelonin induce transformation of fibroblasts, possibly as a consequence of damage to DNA and of the altered DNA repair system. PMID- 12633876 TI - EST analysis of gene expression in the tentacle of Cyanea capillata. AB - Jellyfish, Cyanea capillata, has an important position in head patterning and ion channel evolution, in addition to containing a rich source of toxins. In the present study, 2153 expressed sequence tags (ESTs) from the tentacle cDNA library of C. capillata were analyzed. The initial ESTs consisted of 198 clusters and 818 singletons, which revealed approximately 1016 unique genes in the data set. Among these sequences, we identified several genes related to head and foot patterning, voltage-dependent anion channel gene and genes related to biological activities of venom. Five kinds of proteinase inhibitor genes were found in jellyfish for the first time, and some of them were highly expressed with unknown functions. PMID- 12633877 TI - The dihydroceramide desaturase is not essential for cell viability in Schizosaccharomyces pombe. AB - Recent studies have identified a new family of desaturase-like polypeptide sequences in many higher eukaryotes. Functional characterisation of one member of this family, from Schizosaccharomyces pombe, revealed the enzyme to be a sphingolipid desaturase. This S. pombe gene designated SDCB3b8.07c was identified as the dihydroceramide Delta(4)-desaturase, responsible for the synthesis of sphingosine. Homologous recombination was used to disrupt the endogenous S. pombe dihydroceramide Delta(4)-desaturase. Surprisingly, this had no effect on cell viability, indicating that sphingosine may not be crucial for normal S. pombe functions. This observation has implications for our understanding of the role of sphingosine and its phosphorylated metabolite sphingosine-1-phosphate in lower eukaryotes. PMID- 12633879 TI - Signal peptide of Lassa virus glycoprotein GP-C exhibits an unusual length. AB - Lassa virus glycoprotein is synthesized as precursor GP-C into the lumen of the endoplasmic reticulum and cleaved posttranslationally into the N-terminal subunit GP-1 and the C-terminal subunit GP-2 by subtilase SKI-1/S1P. The N-terminal portion of the primary translation product preGP-C contains a signal peptide of unknown length. In order to demonstrate the signal peptide cleavage site, purified viral GP-1 isolated from Lassa virus particles was N-terminally sequenced as TSLYKGV, identical to amino acids 59-65 of GP-C. Mutational analysis of the amino acid residues flanking the putative cleavage site led to non cleavable preGP-C indicating that no other signal peptide cleavage site exists. Interestingly, GP-C mutants with a non-cleavable signal peptide were not further processed by SKI-1/S1P. This observation suggests that the signal peptide cleavage is necessary for GP-C maturation and hence for Lassa virus replication. PMID- 12633878 TI - A novel protein phosphatase 2C family member (PP2Czeta) is able to associate with ubiquitin conjugating enzyme 9. AB - In this study we have cloned a novel member of mouse protein phosphatase 2C family, PP2Czeta, which is composed of 507 amino acids and has a unique N terminal region. The overall similarity of the amino acid sequence between PP2Czeta and PP2Calpha was 22%. On Northern blot analysis PP2Czeta was found to be expressed specifically in the testicular germ cells. PP2Czeta expressed in COS7 cells was able to associate with ubiquitin conjugating enzyme 9 (UBC9) and the association was enhanced by co-expression of small ubiquitin-related modifier 1 (SUMO-1), suggesting that PP2Czeta exhibits its specific role through its SUMO induced recruitment to UBC9. PMID- 12633880 TI - Early changes in gene expression in two models of Batten disease. AB - Infantile and juvenile neuronal ceroid lipofuscinosis (NCLs) are progressive neurodegenerative disorders of childhood with distinct ages of clinical onset, but with a similar pathological outcome. Infantile and juvenile NCL are inherited in an autosomal recessive manner due to mutations in the CLN1 and CLN3 genes, respectively. Recently developed Cln1- and Cln3-knockout mouse models share similarities in pathology with the respective human disease. Using oligonucleotide arrays we identified reproducible changes in gene expression in the brains of both 10-week-old Cln1- and Cln3-knockout mice as compared to wild type controls, and confirmed changes in levels of several of the cognate proteins by immunoblotting. Despite the similarities in pathology, the two mutations affect the expression of different, non-overlapping sets of genes. The possible significance of these changes and the pathological mechanisms underlying NCL diseases are discussed. PMID- 12633881 TI - Slowing of rat diaphragm action potential depolarization by endurance treadmill training. AB - This study tested the hypothesis that the action potential properties of the diaphragm muscle are altered by endurance exercise treadmill training. Rats underwent treadmill running or sham training for 8 weeks, and intracellular electrophysiological recordings were subsequently performed in vitro. Diaphragm resting membrane potential was not altered by training. The maximal rate of action potential depolarization was reduced significantly by exercise training, from 551+/-16 to 445+/-15 mV/ms (P<0.00002). In contrast the rate of action potential repolarization was not significantly different between the two groups (P=0.25). Action potential height was significantly higher in control compared with trained muscle (84.5+/-1.0 vs. 78.4+/-1.2 mV, P<0.0005). The combination of slowed action depolarization and decreased peak action potential height resulted in no net change in action potential area. Thus treadmill running endurance exercise training slows rat diaphragm action potential depolarization but not repolarization, suggestive of altered Na+ but not K+ channel function. PMID- 12633882 TI - Aging and saccade-stepping interactions in humans. AB - Saccade-stepping interactions were compared in older and young persons to determine if visual anchoring theory could be applied to stepping behavior in both groups. Subjects rapidly stepped onto a platform. In some trials, a translucent shield was used to prevent visual fixation. Both elderly and younger subjects initiated downward saccades prior to footlift, but saccade-step latency was significantly longer for the elderly. Regardless of age, working memory was not sufficient to guide stepping in the absence of visual fixation. The findings support visual anchoring theory by demonstrating that older and young persons generate anticipatory downward saccades which precede the stepping action. Changes in the latency of saccade-step coupling with aging were attributed to slowed processing of visual anchors for obstacle avoidance. The central nervous system appears to accommodate age-related processing delays by initiating the down saccade well in advance of platform footlift, independently from stepping velocity. PMID- 12633883 TI - Aging-related down-regulation of neprilysin, a putative beta-amyloid-degrading enzyme, in transgenic Tg2576 Alzheimer-like mouse brain is accompanied by an astroglial upregulation in the vicinity of beta-amyloid plaques. AB - Pathological accumulation of cortical beta-amyloid is an early and consistent feature of Alzheimer's disease. Brain level of beta-amyloid is determined both by its production and by its catabolism. Neprilysin, a zinc metalloproteinase has been suggested as potential candidate of beta-amyloid-degrading enzyme in vivo. To address the question whether pathological accumulation of beta-amyloid peptides in transgenic Tg2576 mice with Alzheimer-like pathology may affect beta amyloid catabolism, the expression of neprilysin was studied during postnatal maturation and aging. Neprilysin protein but mRNA levels decreased in mouse cerebral cortex with age (2-22 months), independently of transgene status. Immunocytochemistry revealed few neprilysin-positive dystrophic neurites around beta-amyloid plaques and an upregulation of neprilysin in plaque-surrounding reactive astrocytes which may suggest a role of plaque-mediated astrogliosis in beta-amyloid degradation. PMID- 12633884 TI - Effects of aniracetam on extracellular levels of transmitter amino acids in the hippocampus of the conscious gerbils: an intracranial microdialysis study. AB - The effects of aniracetam on extracellular amino acid levels in the hippocampus of conscious gerbils, with or without transient cerebral ischemia/reperfusion, were measured by microdialysis and reverse phase-high performance liquid chromatography. Increased extracellular levels of aspartate and glutamate that were observed in the hippocampus of conscious gerbils during transient global forebrain ischemia were reversed by aniracetam. In contrast, the level of extracellular gamma-aminobutyric acid was increased, while taurine was maintained at a higher level than other amino acids by administration of aniracetam (100 mg/kg, p.o.) 60 min before ischemia. Further, in contrast to ischemic animals, administration of aniracetam (100 mg/kg, p.o.) enhanced the release of glutamate and aspartate in the normal gerbil hippocampus. The results suggest that these effects might be due to a partial calcium agonist activity of aniracetam, and that the effects of aniracetam on amino acid levels might be a mechanism of protection against delayed neuronal death in the ischemic hippocampus, thereby improving memory dysfunction induced by ischemia/reperfusion. PMID- 12633885 TI - Talker's voice and gender stereotype in human auditory sentence processing- evidence from event-related brain potentials. AB - The present study investigated the influence of implicit speaker information on the sentence interpretation. We auditorily presented sentences that comprised of either stereotypically male or stereotypically female self-referent utterances. In the congruent conditions, these utterances were produced by speakers whose gender matched the semantic content. In the incongruent condition, stereotypically male utterances were produced by female speakers and vice versa. The event-related brain potentials (ERP) of 32 listeners exhibited a late positivity (P600) for the incongruent condition. No significant differences were observed between male and female listeners. In the absence of any ERP effect in the earlier time range, it was concluded that the access of the semantic information as such is independent of the speaker's voice, but that speaker property, semantic content and stereotypical knowledge are integrated in a later processing stage. PMID- 12633886 TI - Serum free and phospholipid-bound choline decrease after surgery and methylprednisolone administration in dogs. AB - We designed this study to determine whether serum free and phospholipid-bound choline concentrations change after surgery or methylprednisolone treatment in dogs and rats. In dogs, serum free and phospholipid-bound choline concentrations were decreased by 29% and 17% immediately after abdominal-pelvic surgery under xylasine+ketamine anesthesia, respectively, and both remained low for 24 h. Serum cortisol was elevated after surgery. The elevation in serum cortisol was inversely correlated with the decreases in free (r=-0.737; P<0.001) and phospholipid-bound (r=-0.771; P<0.001) choline concentrations. After methyprednisolone administration (5-20 mg/kg) free and phospholipid-bound choline concentrations decreased in a dose- and time-dependent manner. In rats, either surgery or methylprednisolone failed to alter serum free choline concentrations while phospholipid-bound choline decreased after surgery. These data show that the decrease in serum choline after surgery results from the increase in circulating glucocorticoids. PMID- 12633887 TI - An integrin binding peptide reduces rat CA1 hippocampal long-term potentiation during the first few minutes following theta burst stimulation. AB - The integrin binding peptide GRGDSP was tested on Schaffer to CA1 (Sch-CA1) long term potentiation (LTP) in the rat hippocampal slice. Experiments in which GRGDSP was bath applied for 50 min, beginning 10 min before theta burst stimulation (TBS), reduced LTP of the field excitatory post synaptic potential in a concentration dependent manner, with 250 microM producing a significant decrease. However, LTP was not affected when 250 microM GRGDSP was applied 30 min post-TBS, nor when applied as soon as 5 min post-TBS. When 250 microM GRGDSP was applied for only 10 min pre- to 5 min post-TBS, this brief application was sufficient in reducing LTP similar to the extended 50 min application. We conclude that RGD binding integrins involved in LTP are only momentarily responsive to peptide mediated antagonism in the first few minutes following TBS. PMID- 12633888 TI - Intracerebroventricular injection of vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide inhibits feeding in chicks. AB - Previous research has indicated an involvement of glucagon superfamily peptides in the regulation of feeding in the domestic chick brain. However the possible roles of vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase activating polypeptide-38 (PACAP) have not yet been investigated. We therefore examined the effect of intracerebroventricular (ICV) injections of VIP or PACAP on food intake in chicks. ICV injection of both VIP and PACAP significantly inhibited food intake over 4 h at doses ranging from 12 to 188 pmol. Subsequently, we compared the anorexic effect the glucagon superfamily peptides VIP, PACAP, growth hormone-releasing factor (GRF) and glucagon-like peptide-1 (GLP-1) after ICV injection at an equimolar dose (12 pmol). All four peptides significantly inhibited food intake, although the anorexic effects of VIP and PACAP were weaker than those of GRF and GLP-1. These findings support the hypothesis that glucagon superfamily peptides play an important role in the regulation of appetite in the chick brain. PMID- 12633889 TI - Activation of brown adipose tissue thermogenesis increases slow wave sleep in rat. AB - Considering the thermoregulatory role of slow wave sleep (SWS), we wondered whether the sole increase of brown adipose tissue (BAT) thermogenesis could enhance this sleep state. We tested this hypothesis by administering to rats an agonist (BRL 37,344) of the beta-3 adrenoceptor subtype that is massively localized in BAT cell membrane and that is known to activate BAT thermogenesis. Sleep was electrographically characterized. The temperature of interscapular BAT (Tibat) and cortex (Tco) were also assessed. Tibat significantly increased 2-3 h after BRL injection (but not Tco), concomitantly with SWS (+56-57%). At the maximum of Tibat, a significant positive correlation was found between their changes and those of SWS. We demonstrated for the first time that sleep (and especially SWS) can be affected by the specific activation of BAT. PMID- 12633890 TI - Direct synaptic connections between corticothalamic axon terminals and interneurons in the cat ventrobasal complex. AB - Lesion-induced degeneration was combined with immunocytochemistry to study, with electron microscopy, the synaptic connectivity between corticothalamic axon terminals from the first and second somatosensory areas and local circuit neurons of the ipsilateral ventrobasal complex (VB), selectively labelled with an antibody raised against gamma-aminobutyric acid (GABA). Four days from the cortical ablation many degenerating axon terminals, forming asymmetric synapses, were found on dendritic trees of both labelled and unlabelled neurons of VB and occasionally on presynaptic dendrites. The main finding of the present paper is that 64.01% of degenerating axon terminals synapsed with GABA-immunopositive dendrites, suggesting that the principal target of the cortical projection to VB are local circuit neurons. PMID- 12633891 TI - Accumulation of the proinflammatory cytokine endothelial-monocyte-activating polypeptide II in ramified microglial cells in brains of Borna virus infected Lewis rats. AB - Borna disease virus (BDV) infection of adult Lewis rats induces a severe and often fatal neurologic disease characterized by a massive mononuclear meningo encephalitis, and activation of microglial cells. Therefore, we analyzed expression of endothelial monocyte activating polypeptide II (EMAP II) by immunohistology as a marker for activation of microglial cells in BDV infected rat brains. EMAP II is a chemotactic peptide, inducing activation of macrophages and endothelial cells, and is considered a proinflammatory mediator of the innate immune system. An up to 30-fold increase in numbers of EMAP II+ microglial cells and a massive expression by infiltrating macrophages at perivascular inflammatory foci was observed in infected brains, with a maximum on day 25 after infection. These results provide evidence that EMAP II contributes to immune responses in inflammatory processes caused by viral infections. PMID- 12633892 TI - Hippocampal CA1 neuron survival and cytosolic FKBP12, the 12 kDa FK506-binding protein, after ischemia and tacrolimus treatment in gerbils. AB - To assess the neuroprotective effect of tacrolimus (FK506) in transient forebrain ischemia models in gerbils, 10.0 mg/kg of FK506 was injected intraperitoneally immediately following reperfusion and at intervals of 1, 3, 6, 9, and 12 h after reperfusion. FK506 produced a significant neuroprotective effect for up to 6 h after 5 min of ischemia. Immunoblot and immunohistochemistry revealed that the amount of FKBP12, the 12-kDa FK506-binding protein, in the cytosol remained unchanged until 12 h after reperfusion. Translocation of FKBP12 from the nucleus to the cytosol was not observed until 24 h after reperfusion. Administration of FK506 did not appear to induce the cytosolic increase in FKBP12. In this study, no correlation was apparent between the post-ischemic therapeutic efficacy of FK506 and the post-ischemic changes in the cytosolic FKBP12. PMID- 12633893 TI - Influence of a functional polymorphism within the angiotensin I-converting enzyme gene on partial sleep deprivation in patients with major depression. AB - Partial sleep deprivation (PSD) exerts transient antidepressant efficacy. As a potential mechanism of action an enhancement of dopaminergic neurotransmission within the CNS is discussed. Because genetic variations influencing neurotransmission could have an impact on therapeutic outcome and stability of improvement, we investigated the functional insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme (ACE) gene to examine a possible influence on the dopaminergic pathway. We included 56 patients with major depression (DSM IV). Psychiatric ratings including the HAM-D6 scale were assessed prior to and after PSD and related to the different genotypes. The ACE I/D polymorphism was determined following PCR amplification using genomic DNA. A total of 58.1% of the patients were PSD responders. As expected, the therapeutical effect of PSD was transient and most patients experienced an exacerbation of depressive symptoms on day 2. Subdivision according ACE gene variants showed a significantly less pronounced relapse of symptoms in ACE gene D-allele carriers (P=0.02). Our results give first hints that the ACE I/I genotype, probably influencing dopaminergic neurotransmission, could be an indicator for relapse after PSD. This should result in earlier and more intense additional therapeutic interventions in this group of patients. PMID- 12633894 TI - Effects of repeated stress on regeneration of serotonergic and noradrenergic axons in the cerebral cortex of adult rats. AB - We examined whether regeneration of serotonergic (5-HT) and noradrenergic (NA) axons might be affected by stress. Neurotoxins to 5-HT or NA axons were injected into the frontal cortex to cause partial denervation. Mild restraint stress (40 min/day) was started 16 days later and given for 14 consecutive days (30-day stressed group). Non-stressed animals were divided into two groups, animals sacrificed at 14 days (14-day control group) and those sacrificed at 30 days (30 day control group) after the toxin injection. In immunohistochemical study, the denervation area of 5-HT axons but not that of NA axons was significantly smaller in the 30-day control group than in the 14-day control group. However, there was no significant difference between the 14-day control and 30-day stressed groups. These findings suggested that regeneration of 5-HT axons occurred earlier than that of NA axons, and that stress exerted inhibitory influence on regeneration of 5-HT axons. PMID- 12633895 TI - Neurotrophins facilitate synthesis of choline acetyltransferase and tyrosine hydroxylase in cultured mouse neural stem cells independently of their neuronal differentiation. AB - Effects of three neurotrophins, i.e., nerve growth factor, brain-derived neurotrophic factor, and neurotrophin-3, on the expression of four neurotransmitter-synthesizing enzymes, i.e. choline acetyltransferase (ChAT), tyrosine hydroxylase (TH), dopamine beta hydroxylase (DBH), and glutamate decarboxylase 65 were investigated in cultured mouse neural stem cells. All three neurotrophins enhanced the mRNA expression of ChAT, TH, or DBH of the cells caused to differentiate by the removal of fibroblast growth factor (FGF)-2 from the culture medium, and increased the protein and mRNA levels of ChAT and TH of even the undifferentiated proliferating neural stem cells due to the presence of FGF-2. These results demonstrate that neurotrophins stimulate the synthesis of ChAT and TH of the neural stem cells prior to neuronal differentiation, and suggest that neurotrophins may play roles in the commitment to neuronal cells and choice of specific neurotransmitter phenotypes in early stages of neurogenesis. PMID- 12633896 TI - Serotonin produces an enhanced outward current recorded at rat dorsal lateral septal neurons from the Flinders Sensitive Line of rats, a genetically-selected animal model of depression. AB - Abnormalities in serotonin (5-HT), serotonin receptors, and serotonergic neurons have been reported in studies of brains from patients diagnosed clinically with depression. In this study, we examined a known cellular function of 5-HT(1A) receptor activation in dorsolateral septal nucleus (DLSN) neurons, namely, a concentration dependent 5-HT-induced outward current, and compared basic neuronal membrane properties and activities of DLSN neurons from two known genetic lines of rats. As compared to "control" rats (Flinders Resistant Line, FRL), DLSN neurons from Flinders Sensitive Line of rats (FSL) did not exhibit significant differences in resting membrane potential, membrane input resistance, or changes in typical spontaneous inhibitory or excitatory post-synaptic currents. FSL-rats exhibit a depressive phenotype and have been suggested to be rats with a genetic susceptibility to exhibit depressive behaviors. Exogenous application of 5-HT resulted in expected concentration-dependent outward currents; however, the amplitudes of these currents were enhanced significantly in 50% of DLSN neurons recorded from FSL rats compared to similar results recorded from FRL rats. Our results suggest that within a particular population of DLSN neurons from rats exhibiting a known phenotype of depression a post-synaptic 5-HT(1A) receptor is functionally hyper-responsive compared to controls. PMID- 12633897 TI - Galanin attenuates basal and antidepressant drug-induced increase of extracellular serotonin and noradrenaline levels in the rat hippocampus. AB - Galanin is co-localized with classical neurotransmitters, such as acetylcholine, serotonin (5-HT) and noradrenaline (NA) in neurons or in brain regions implicated in cognitive and affective behaviour. In the present study, the effects of galanin on extracellular 5-HT and NA levels in the rat hippocampus were measured by in vivo microdialysis under basal conditions and following systemic administration of antidepressant drugs. Galanin (1.5 nmol i.c.v.) reduced basal 5 HT and NA levels to 65% and 86% of controls, respectively. Galanin (0.5 and 1.5 nmol i.c.v.) dose-dependently attenuated the elevation of 5-HT concentrations induced by imipramine and citalopram (10 mg/kg i.p., each) from 350% to 312% and from 230% to 160%, respectively. Galanin at 1.5 nmol transiently attenuated the effect of desipramine-induced (10 mg/kg i.p.) increase in extracellular NA levels from a maximal increase of 389-296% of the predrug levels. It is concluded that intraventricularly administered galanin attenuated both basal 5-HT and NA release and antidepressant drug-induced accumulation of extracellular 5-HT and NA levels most likely via a predominant inhibitory action on serotonergic and noradrenergic neurons in the raphe and locus coeruleus, respectively. These results further emphasize a possible role of galanin in regulation of 5-HT and NA neurotransmission in depressive states and during the course of antidepressant therapy. PMID- 12633898 TI - Spatial invariance in anticipatory orienting behaviour during human navigation. AB - We have recently reported that the head systematically deviates toward the future direction of the trajectory about 500 ms before attaining a turning point of 90 degrees corner trajectories both in light and in darkness. Here, we investigated how this anticipatory strategy is modified whilst varying visual conditions (Experiment 1) and walking speed (Experiment 2). Exp. 1 showed similar anticipatory behaviour when walking with or without vision. Exp. 2 (that varied walking speed; eyes open) showed that the head started to deviate at a constant distance rather than at a constant time to the corner. The results appear inconsistent with optic flow theories of the guidance of walking direction and might highlight the role of landmarks and/or egocentric direction in anticipatory orienting behaviour. PMID- 12633899 TI - Neuroprotective effects of aspirin in patients with acute cerebral infarction. AB - Aspirin may reduce ischemic brain injury. The aim of this study was to explore the effect of aspirin on glutamate release after acute stroke. We studied 238 patients with a first episode of hemispheric ischemic stroke of less than 24 h duration. Early neurological deterioration was diagnosed when the Canadian Stroke Scale dropped 1 or more points between admission and 48 h. Glutamate was determined on cerebrospinal fluid (CSF) samples obtained at admission. Sixty three patients were undergoing treatment with 75-500 mg/day of aspirin at the time of stroke onset. CSF glutamate concentrations were higher in the group of patients not taking aspirin (8.9+/-5.2 vs. 4.9+/-3.1 microM/l, P< 0.0001). Aspirin treatment at stroke onset had a 97% risk reduction of early neurological deterioration, and this effect remained unchanged after a further adjustment for glutamate concentrations. These findings suggest that low doses of aspirin may be useful in the management of patients with cerebral ischemia, not only for its antithrombotic properties, but also by direct neuroprotective effects. PMID- 12633900 TI - Cyclosporine A inhibits acetylcholinesterase activity in selected parts of the rat brain. AB - Cyclosporine A (CsA) is the major immunosuppressive drug used for organ and neural transplantation and the therapy of selected autoimmune diseases. We investigated the effect of CsA on the activity of acetylcholinesterase (AChE) in the frontal cortex, hippocampus, septum, and basal ganglia. AChE was determined spectrophotometrically with acetylthiocholine as substrate and 5,5-bis-2 nitrobenzoic acid as chromogen. CsA was administered in single doses of 20 or 45 mg/kg perorally; in the case of the higher dose we also performed a repeated administration of CsA in three consecutive doses separated by 24 h intervals. Both lower and higher doses of CsA decreased AChE activity in the frontal cortex and hippocampus to practically the same extent. On the contrary, AChE activity was more diminished in the case of the higher dose of CsA used in the septum and basal ganglia. Repeated administration of the higher dose of CsA did not lead, with the exception of the hippocampus, to a further decrease in AChE activity in the brain structures observed. These findings contribute to rare evidence concerning the interaction of CsA and the cholinergic system in the brain. PMID- 12633901 TI - Anti-apoptotic proteins are oxidized by Abeta25-35 in Alzheimer's fibroblasts. AB - We have examined the effects of the beta-amyloid peptide (Abeta(25-35)) on fibroblasts derived from subjects with Alzheimer's disease (AD) and from age matched controls. The peptide was significantly more cytotoxic to the AD-derived fibroblasts. The level of protein oxidation was also greater in the cells from AD subjects. Two-dimensional electrophoresis (2-DE) coupled with immunostaining for protein carbonylation revealed specific oxidation-sensitive proteins (OSPs) in both the control and AD-derived cells. Two specific OSPs were identified by mass spectrometry as heat shock protein 60 (HSP 60) and vimentin. Exposure of the cells to Abeta(25-35) resulted in a twofold increase in the level of oxidation of these two OSPs in the cells derived from controls, but a ninefold increase in their level of oxidation in the fibroblasts from AD subjects. These observations are of particular interest because of the proposed anti-apoptotic roles of both HSP 60 and vimentin and our recent observation that these same two proteins are particularly susceptible to oxidation in neuronally derived cells. PMID- 12633902 TI - Oxidant stress is a significant feature of primary biliary cirrhosis. AB - Primary biliary cirrhosis (PBC) is a chronic cholestatic disorder characterised by an immunological, and often granulomatous, attack on bile ducts leading to fibrosis, cirrhosis, liver failure and death. Animal and human studies suggest that oxidant stress plays a key role in progression of other liver diseases, but no comprehensive investigation has been performed previously in PBC. A wide range of lipid peroxidation and antioxidant markers were measured in the blood and urine of 41 patients with histologically confirmed PBC. Lipid peroxidation markers were significantly elevated [plasma and urinary 8-isoprostane, P<0.001; plasma malondialdehyde (MDA), P=0.007] compared to age- and sex-matched controls. The most striking antioxidant depletion occurred with plasma total glutathione where levels were significantly reduced (30% of controls). Total serum antioxidant levels were decreased (P=0.013) and serum selenium and vitamin A were also lower (both P<0.001); vitamins C and E were normal. Most patients had early disease biochemically and were Child-Pugh grade A. Urinary 8-isoprostane correlated positively with Ludwig stage and markers of hepatic injury and cholestasis. This study clearly demonstrates that oxidant stress, as reflected in a comprehensive spectrum of lipid peroxidation and antioxidant markers, is a significant feature of early-stage PBC. PMID- 12633903 TI - Oral administration is as effective as intraperitoneal administration of amifostine in decreasing nitroxide EPR signal decay in vivo. AB - A rapid method to determine the systemic incorporation of amifostine has been sought in order to determine the effectiveness of different administration routes without the delay inherent in awaiting therapeutic results. Consistent changes in animal measurements of nitroxide signal decay were monitored using in vivo EPR at frequencies low enough to ensure uniform sensitivity to organs deep in 20-g C3H mice. Conditions included both co-administration of the amifostine with the carbamoyl-proxyl spin probe (CP) via i.p. injection (n=6) and oral administration (n=8) of the amifostine. These decreased the first order rate of decay of the CP EPR signal after a dose of 13.5 Gy radiation, by 23% and 18%, respectively. These changes were significantly different from the rate of decay of the CP EPR signal without amifostine, but were statistically indistinguishable from each other. These data demonstrate: (1) condition-dependent exponential decay of CP EPR signal allowing its use to determine systemic availability of a drug, and (2) that oral administration and i.p. injection of amifostine are both effective in affecting the CP EPR signal decay rate in a mouse model. This is a strong indicator of similar bioavailability in mice from both routes of administration. PMID- 12633904 TI - Sources of glucose production in cirrhosis by 2H2O ingestion and 2H NMR analysis of plasma glucose. AB - Plasma glucose 2H enrichment was quantified by 2H NMR in patients with cirrhosis (n=6) and healthy subjects (n=5) fasted for 16 h and given 2H(2)O to approximately 0.5% body water. The percent contribution of glycogenolysis and gluconeogenesis to glucose production (GP) was estimated from the relative enrichments of hydrogen 5 and hydrogen 2 of plasma glucose. Fasting plasma glucose levels were normal in both groups (87+/-7 and 87+/-24 mg/dl for healthy and cirrhotic subjects, respectively). The percent contribution of glycogen to GP was smaller in cirrhotics than controls (22+/-7% versus 46+/-4%, P<0.001), while the contribution from gluconeogenesis was larger (78+/-7% versus 54+/-4%, P<0.001). In all subjects, glucose 6R and 6S hydrogens had similar enrichments, consistent with extensive exchange of 2H between body water and the hydrogens of gluconeogenic oxaloacetate (OAA). The difference in 2H-enrichment between hydrogen 5 and hydrogen 6S was significantly larger in cirrhotics, suggesting that the fractional contribution of glycerol to the glyceraldehyde-3-phosphate (G3P)-moiety of plasma glucose was higher compared to controls (19+/-6% versus 7+/-6%, P<0.01). In all subjects, hydrogens 4 and 5 of glucose had identical enrichments while hydrogen 3 enrichments were systematically lower. This reflects incomplete exchange between the hydrogen of water and that of 1-R dihydroxyacetone phosphate (DHAP) or incomplete exchange of DHAP and G3P pools via triose phosphate isomerase. PMID- 12633905 TI - Lysosomal dysfunction in muscle with special reference to glycogen storage disease type II. AB - The importance of proper lysosomal activity in cell and tissue homeostasis is underlined by "experiments of nature", i.e. genetic defects in one of the at least 40 lysosomal enzymes/proteins present in the human cell. The complete lack of 1-4 alpha-glucosidase (glycogen storage disease type II (GSD II) or Pompe disease) is life-threatening. Patients suffering from GSD II commonly die before the age of 2 years because of cardiorespiratory insufficiency. Striated muscle cells appear to be particularly vulnerable in GSD II. The high cytoplasmic glycogen content in muscle cells most likely gives rise to a high rate of glycogen engulfment by the lysosomes. The polysaccharides become subsequently trapped in these organelles when 1-4 alpha-glucosidase activity is absent. During the course of the disease, muscle wasting occurs. It is hypothesised that the gradual loss of muscle mass is caused by a combination of disuse atrophy and lipofuscine-mediated apoptosis of myocytes. Moreover, we hypothesise that in the remaining skeletal muscle cells, longitudinal transmission of force is hampered by swollen lysosomes, clustering of non-contractile material and focal regions with degraded contractile proteins, which results in muscle weakness. PMID- 12633906 TI - Polyethylene glycol-conjugated adenosine phosphorylase: development of alternative enzyme therapy for adenosine deaminase deficiency. AB - Purine nucleoside phosphorylase had previously been engineered to accept 6-amino substituted purine nucleosides by two active site substitutions, Asn243Asp; Lys244Gln. In the present study, recombinant adenosine phosphorylase (AP) has been conjugated to branched polyethylene glycol (PEG) polymers of approximately 42.5 kDa. Matrix-assisted laser desorption/ionization analysis and SDS acrylamide electrophoresis analysis indicated a subunit composition of greater than 205 kDa consistent with the conjugation of as many as four PEG molecules per AP subunit. The PEG-conjugated enzyme retained greater than 90% of the native catalytic activity. Administration of the enzyme to mice demonstrated the PEG-AP to have a 67-fold increased plasma half-life compared to the native enzyme, 65.1+/-2.9 h versus 57.8+/-1.1 min, respectively. PEG-AP was principally confined to the plasma with minimal activity detected in tissues and of these spleen had the greatest activity and essentially no activity was found in urine. PEG-AP has retained activity with inosine and its injection into PNP-deficient mice resulted in a 2.7-fold increase in urine urate. AP was also shown to protect human CEM cells in culture from the toxic effects of 2'-deoxyadenosine. These studies provide evidence for consideration of PEG-AP as an alternative enzyme therapy for the inherited deficiency of adenosine deaminase. PMID- 12633907 TI - Cloning and expression of the dihydroorotate dehydrogenase from Toxoplasma gondii. AB - A full-length dihydroorotate dehydrogenase (DHODase) sequence was cloned from a Toxoplasma gondii tachyzoite cDNA library. The sequence was most similar to family 2 DHODases, and had a calculated molecular mass of 65.1 kDa. The full length and two N-terminally truncated T. gondii DHODase sequences were expressed as recombinant proteins. One of the truncated sequences complemented a DHODase deficient bacterial host. PMID- 12633908 TI - College on Problems of Drug Dependence taskforce on prescription opioid non medical use and abuse: position statement. AB - This position paper from the College on Problems of Drug Dependence addresses the issues related to non-medical use and abuse of prescription opioids. A central theme throughout is the need to strike a balance between risk management strategies to prevent and deter prescription opioid abuse and the need for physicians and patients to have appropriate access to opioid pharmaceuticals for the treatment of pain. The epidemiology of prescription opioid use and abuse is reviewed. Non-medical use and abuse of prescription opioids are on the rise in the United States, illicit use of several widely prescribed opioids has increased disproportionately more than illicit use, and the prevalence of prescription opioid abuse appears to be similar to that of heroin and cocaine abuse. There is a paucity of abuse liability testing of prescription opioids, and methods should be developed to fill critical gaps in our knowledge in this area. The role of regulatory agencies in preventing diversion of prescription opioids and identifying potential sources of diversion are discussed. More research is needed to identify those populations most at risk for abusing prescription opioids, and then to develop appropriately targeted prevention programs. Treatment options are discussed; these depend on whether or not an abuser is in pain. Prescription opioid abuse has harmful ramifications for the legitimate and appropriate use of opioids, including stigmatization, opiophobia, and undertreatment of pain. Recommended steps to take include further epidemiological research, laboratory testing of prescription opioids to determine abuse liability, and clinical trials to determine the efficacy of different approaches to the prevention and treatment of prescription opioid abuse. PMID- 12633909 TI - Physical dependence on Ultram (tramadol hydrochloride): both opioid-like and atypical withdrawal symptoms occur. AB - In 1994, the Drug Abuse Advisory Committee (DAAC) of the Food and Drug Administration (FDA) concluded that Ultram (tramadol hydrochloride) could be marketed as an analgesic drug without scheduling under the Controlled Substances Act based upon extensive pre-clinical, clinical and European epidemiological data. However, to guard against unexpectedly high levels of abuse in the United States, the DAAC recommended that an independent steering committee (ISC) be appointed to proactively monitor abuse/dependence. In the event that high rates of abuse were found, this ISC was given the authority to immediately recommend to the FDA that Ultram be scheduled. In the course of the surveillance project, the ISC received reports of withdrawal following abrupt discontinuation of Ultram and in some instances, following dose reductions. In most cases, the withdrawal symptoms consisted of classical opioid withdrawal, but in some cases were accompanied by withdrawal symptoms not normally observed in opiate withdrawal, such as hallucinations, paranoia, extreme anxiety, panic attacks, confusion and unusual sensory experiences such as numbness and tingling in one or more extremities. Withdrawal symptoms of either type were one of the more prevalent adverse events associated with chronic Ultram use, comprising nearly 40% of all adverse events reported with Ultram. Most of these consisted of typical opiate withdrawal symptoms, but 1 in 8 cases presented as atypical. These results indicate that physicians and other healthcare professionals need to be aware of the potential of Ultram to induce withdrawal of the classical opioid type, and that atypical withdrawal may also occur. PMID- 12633910 TI - Predictors of follow-up health status in the Drug Abuse Treatment Outcome Study (DATOS). AB - This study examined the predictors of self-reported health status at follow-up in the Drug Abuse Treatment Outcomes Study (DATOS), a longitudinal study of drug abuse treatment programs and patients in 1991-1993. Baseline and follow-up interviews of 2966 patients in 75 programs were performed. The follow-up assessment was targeted to occur 12 months after treatment terminated; long-term methadone patients in treatment for the entire 12-month period were interviewed 24 months after intake. A composite measure, developed through principal component analysis, assessed health status. A multivariate hierarchical linear regression model adjusted for identified independent baseline predictors of health status at follow-up. Poor physical health status (including the composite measure, comorbid conditions and pain) and greater severity of psychiatric symptoms at baseline were the strongest predictors of poor health status at follow-up. Other predictors of worse health status included older age, public insurance coverage and unemployment. We conclude that baseline health status and psychiatric symptoms predict the subsequent health status of patients in substance abuse treatment patients as in other clinical populations. Future research should examine whether early identification and treatment of physical and mental health problems among patients in addiction treatment programs might remediate their adverse effects on long-term health status outcomes. PMID- 12633911 TI - The association between parental alcohol-related behaviors and children's drinking. AB - Past literature has established an association between children's alcohol-related behaviors and parents' use of and attitudes toward alcohol. However, most studies tend to measure parental use and attitudes through proxy, i.e. children's perception. It is not clear the extent to which actual parental use and attitudes influence children's alcohol behaviors. The current study directly used parents' reports on alcohol use and attitudes toward alcohol and examined their impact on children's alcohol use. Based on a three-stage random sampling design, 642 dyads of parents and children (ages 15-18) were interviewed by telephone in New York State. Study variables include parental alcohol use, children's alcohol use, parental attitudes toward underage drinking and parent-child interaction. While parental use and attitudes do not seem to significantly affect children's alcohol use, the extent to which parents prohibit children from using alcohol at home tends to reduce children's alcohol involvement. In addition, the greater the amount of time spent with alcohol-using parents, the more likely the children are to use alcohol. Findings suggest that, while parents' alcohol use influences children's alcohol use through extensive interaction, parental control of underage alcohol use in the household appears to reduce children's involvement in underage alcohol use. PMID- 12633913 TI - Association of cocaine withdrawal symptoms with more severe dependence and enhanced subjective response to cocaine. AB - The purpose of this two part study was to better characterize cocaine users based on self-reported cocaine withdrawal symptoms by examining screening data and response to smoked cocaine in the human laboratory. The first study sample included male and female non-treatment seeking cocaine users who were screened as potential subjects for inpatient studies. Of the 555 subjects, 462 (82%) endorsed symptoms consistent with DSM-IV criteria for cocaine withdrawal. Cocaine users who met criteria for cocaine withdrawal, compared with those who did not, reported a significantly higher amount of cocaine use and a history of medical and psychosocial problems. Cocaine users meeting DSM-IV withdrawal criteria, which included endorsement of depression, were also more likely to have a history of depression, to have seriously considered suicide, and to have had chemical dependency treatment even when amount spent on cocaine was covaried. The second study sample, which was a subset of Study I, included those who participated in human cocaine studies following the phone screening. Cocaine users who met criteria for cocaine withdrawal (n=34), compared with those who did not (n=10), had enhanced subjective ratings of 'high' and 'feel the effect of last dose' in response to a single delivery of 0.4 mg/kg of smoked cocaine. These results suggest that history of cocaine withdrawal symptoms may be associated with enhanced cocaine responses and greater severity of cocaine dependence. PMID- 12633912 TI - Opioid detoxification with buprenorphine, clonidine, or methadone in hospitalized heroin-dependent patients with HIV infection. AB - With the growing role of intravenous drug use in the transmission of HIV infection, HIV-infected patients frequently present with comorbid opioid dependence. Yet, few empirical evaluations of the efficacy and consequences of opioid detoxification medications in medically ill HIV-infected patients have been reported. In a randomized, double-blind clinical trial, we evaluated the impact of three medications on the signs and symptoms of withdrawal and on the pain severity in heroin-dependent HIV-infected patients (N=55) hospitalized for medical reasons on an inpatient AIDS service. Patients received a 3-day pharmacologic taper with intramuscular buprenorphine (n=21), oral clonidine (n=16), or oral methadone (n=18), followed by a clonidine transdermal patch on the fourth day. Observed and self-reported measures of opioid withdrawal and pain were taken 1-3 times daily for up to 4 days. Opiate administration used as medically indicated for pain was also recorded. Observer- and subject-rated opiate withdrawal scores decreased significantly following the first dose of medication and overall during treatment. Among all 55 subjects, self-reported and observer-reported pain decreased after treatment (on average observer-rated opioid withdrawal scale (OOWS) scores declined 5.6 units and short opioid withdrawal scale (SOWS) declined 4.8 units, P<0.001, for both) with no indication of increased pain during medication taper. There were no significant differences of pain decline and other measures of withdrawal between the three treatment groups. During the intervention period, supplemental opiates were administered as medically indicated for pain to 45% of the patients; only 34% of men versus 62% of women received morphine (P<0.05). These findings suggest buprenorphine, clonidine, and methadone regimens each decrease opioid withdrawal in medically ill HIV-infected patients. PMID- 12633914 TI - Ultra-rapid opiate detoxification using deep sedation and prior oral buprenorphine preparation: long-term results. AB - BACKGROUND: New methods of ultra-rapid opiate detoxification (URD) under intravenous sedation have been criticized because of limited data on safety and long-term follow-up. Premedication with buprenorphine has been advocated to improve safety by decreasing vomiting. Prior research has not explored URD in socially impaired patients. METHOD: Sixteen patients were detoxified with URD and prospectively evaluated over at least 30 months. Data of this procedure were compared with those of our previous study without buprenorphine preparation (Drug Alcohol Depend. 52(3) (1998) 243). The 16 patients were followed up by a general practitioner (GP) before and after URD. The GPs also supervised the 7-day course of buprenorphine treatment prescribed for the 16 patients prior to URD. RESULTS: During the procedure, only one episode of vomiting occurred instead of 13 out of 20 in our previous study. Post-procedure, only two patients experienced moderate withdrawal symptoms, such as persistent nausea, abdominal cramps and vomiting lasting from 24 to 48 h, in comparison with most patients in the previous study without buprenorphine. After a period of at least 30 months (36.0+/-6.38), the 16 patients were still alive and were regularly monitored by their GP. Only two of the 16 never relapsed after URD and reported total opiate abstinence. Fourteen patients relapsed; 12 of these were prescribed a licensed methadone substitution program and two were still using heroin. CONCLUSION: In this small sample, the data indicated that URD with buprenorphine preparation was safe and that it markedly decreased post-procedure morbidity. No patient died over a minimum 30 month follow-up period. Furthermore, the procedure was employed with socially impaired patients. In the long term, a few patients were still free of opiates, while the majority opted for a methadone maintenance program, showing that URD can serve as one possible step in a long-term treatment program. PMID- 12633915 TI - Choosing to take cocaine in the human laboratory: effects of cocaine dose, inter choice interval, and magnitude of alternative reinforcement. AB - Cocaine abuse involves a variety of behaviors including the initiation of cocaine seeking, the self-selected patterning of cocaine administrations, and the cessation of cocaine-taking. To date, most human laboratory models of cocaine self-administration have only assessed the amount of cocaine consumed under a fixed set of conditions. This double-blind, randomized, within-subject, inpatient study evaluated a novel model of human cocaine self-administration that aimed to quantify the reinforcing value of cocaine after cocaine-taking was initiated. Cocaine-dependent volunteers (n=8) sampled cocaine (12.5, 25 or 50 mg per 70 kg i.v.) or placebo and were subsequently allowed to choose between another injection of the same dose or money over six trials during 12 experimental sessions. The value of the monetary alternative increased with each trial from US dollars 1 to 16. Each cocaine dose was assessed under three inter-choice intervals: 15 min, 30 min, and an interval selected by the volunteer. Injection choices increased dose dependently; however, there was little relationship between the value of the alternative reinforcer and the choice to take cocaine. Most volunteers exclusively chose injections when active cocaine was available and money when placebo was available. Inter-choice interval did not affect cocaine choices. These results illustrate the persistence of cocaine self administration once cocaine-taking has been initiated. PMID- 12633916 TI - Early-onset cannabis use and cognitive deficits: what is the nature of the association? AB - BACKGROUND: Individuals who initiate cannabis use at an early age, when the brain is still developing, might be more vulnerable to lasting neuropsychological deficits than individuals who begin use later in life. METHODS: We analyzed neuropsychological test results from 122 long-term heavy cannabis users and 87 comparison subjects with minimal cannabis exposure, all of whom had undergone a 28-day period of abstinence from cannabis, monitored by daily or every-other-day observed urine samples. We compared early-onset cannabis users with late-onset users and with controls, using linear regression controlling for age, sex, ethnicity, and attributes of family of origin. RESULTS: The 69 early-onset users (who began smoking before age 17) differed significantly from both the 53 late onset users (who began smoking at age 17 or later) and from the 87 controls on several measures, most notably verbal IQ (VIQ). Few differences were found between late-onset users and controls on the test battery. However, when we adjusted for VIQ, virtually all differences between early-onset users and controls on test measures ceased to be significant. CONCLUSIONS: Early-onset cannabis users exhibit poorer cognitive performance than late-onset users or control subjects, especially in VIQ, but the cause of this difference cannot be determined from our data. The difference may reflect (1). innate differences between groups in cognitive ability, antedating first cannabis use; (2). an actual neurotoxic effect of cannabis on the developing brain; or (3). poorer learning of conventional cognitive skills by young cannabis users who have eschewed academics and diverged from the mainstream culture. PMID- 12633917 TI - Prevalence and correlates of areca nut use among psychiatric patients in India. AB - OBJECTIVES: To estimate the prevalence and identify the correlates of areca nut use among men and women being treated for a major psychiatric disorder in India. METHODS: Inpatients (N=988) admitted to the adult psychiatry department of the National Institute of Mental Health and Neuro Sciences in India were interviewed regarding their use of the areca nut, tobacco, alcohol, and other drugs. Medical records were reviewed to obtain psychiatric diagnosis and history. RESULTS: About 24% of the sample reported recent areca nut use, and 10% reported severe use suggesting dependence. Common reasons for use include to improve mood (31% of users), socialization (31%), digestion (22%), or performance (7%) and to decrease aches and pains (6%). Predictors of current areca nut use included less education, diagnosis of bipolar disorder, and current tobacco use. Predictors of severe use were older age, female gender, less education, and current tobacco use. CONCLUSION: Areca nut use occurs commonly among Indian psychiatric patients, and deserves further investigation. PMID- 12633919 TI - Embryonic stem cells enhance the healing of tympanic membrane perforations. AB - OBJECTIVE: Tympanic membrane perforations may cause hearing impairment and otorhea. It is a common indication for ear surgery. The aim of the study was to test whether stem cells may enhance the healing of fresh tympanic membrane perforations. METHODS: In a first assay, the status of the tympanic membrane at 5 days after myringotomy was tested in five Mongolian gerbils that were treated on one side with embryonic stem cells and on the other side with control substance. In a second assay, nine gerbils were treated in the same way, except that fluorescent-labeled embryonic stem cells were used. The integration of the stem cells into the surface layer of the healing tympanic membrane was assessed with fluorescence microscopy, as well as the differentiation of these cells. RESULTS: In the first assay, all perforations in the treated ears were closed, whereas only two of the untreated ears were closed. The strength of the healed perforation was greater in the stem cell treated tympanic membranes (mean rupture pressure 120 daPa in three treated ears compared to 60 daPa in the one control ear). Two stem cell-treated tympanic membranes remained intact throughout the whole sequence of pressures, whereas only one control tympanic membrane remained intact. In three tympanic membranes in the second assay, a group of fluorescence doped cells was detected in the region of the perforation. CONCLUSION: These findings indicate that stem cells enhance the healing of tympanic membrane perforations, possibly by differentiation and integration into the tympanic membrane tissue. PMID- 12633918 TI - Opioid effects and opioid withdrawal during a 24 h dosing interval in patients maintained on buprenorphine. AB - In maintenance patients methadone has been shown to produce considerable changes in opioid effects and withdrawal over the dosing interval. As a partial agonist buprenorphine may be expected to produce smaller changes, but the nature and magnitude of these changes have only been described for single doses. In the present study opioid effects and withdrawal were described in patients maintained on buprenorphine. Twenty four opioid dependent subjects were administered 16 mg buprenorphine tablets sublingually for 10 days. On day 10 plasma samples were collected and physiological, subjective and observer-rated measures collected pre dose and at 14 time points during the dosing interval. No significant respiratory depression was observed. Consistent with the partial agonist properties of buprenorphine, other physiological and subjective changes were also of small magnitude. However, even at a once daily dose of 16 mg some patients experienced significant opioid withdrawal that was maximal at the end of the dosing interval. Buprenorphine maintenance should be associated with a high level of safety and a low level of disruption caused by changing opioid effects over the dosing interval, but some patients may require high doses or other strategies to completely suppress withdrawal. PMID- 12633920 TI - Middle ear fluid histamine and leukotriene B4 in acute otitis media: effect of antihistamine or corticosteroid treatment. AB - OBJECTIVE: Two potent mediators of acute inflammation, histamine and leukotriene B4 (LTB4), have been shown to play important roles in the pathogenesis and clinical course of acute otitis media (AOM) in children. The purpose of this study was to evaluate the ability of adjuvant drugs, antihistamine and corticosteroid, in reduction of the levels of histamine and LTB4 in the middle ear and their ability to improve outcomes of AOM. METHODS: Eighty children with AOM (aged 3 months to 6 years) were enrolled in a prospective, randomized, double blind, placebo controlled study. All children received one dose of intramuscular ceftriaxone and were randomly assigned to receive either chlorpheniramine maleate (0.35 mg/kg per day) and/or prednisolone (2 mg/kg per day) or placebos three times a day for 5 days. Tympanocentesis was performed at enrollment and after 5 days of adjuvant drug treatment. MEFs were collected for bacterial and viral studies and histamine and LTB4 levels. The subjects were followed for the duration of middle ear effusion or up to 3 months. RESULTS: Histamine or LTB4 levels in the MEF after 5 days of treatment were not significantly reduced by adjuvant drug treatment. However, subjects receiving corticosteroid had a lower rate of treatment failure during the first 2 weeks and shorter duration of middle ear effusion. CONCLUSIONS: Five day of antihistamine or corticosteroid treatment does not reduce the levels of histamine or leukotriene B4 in the MEF of children with AOM. Positive clinical outcomes of AOM cases associated with corticosteroid treatment needs to be confirmed in a larger clinical trial of children with intact tympanic membranes, who do not receive tympanocentesis. PMID- 12633921 TI - Congenital middle ear cholesteatoma: need for early recognition--role of computed tomography scan. AB - BACKGROUND: Congenital cholesteatoma (CC) of middle ear is a rare entity that may go undiagnosed for years. Patients with CC who are diagnosed at a later stage of disease have poor outcome. There is controversy regarding the best way to delineate CC preoperatively. More specifically, the need to obtain preoperative computed tomography (CT) scan in all cases of CC is debated. OBJECTIVES: This study was conducted to determine factors that may influence the outcome of surgery in CC as well as the value of obtaining preoperative CT scan in CC. METHOD: A retrospective chart review of all patients with a diagnosis of middle ear cholesteatoma operated on between 1994 and 2000 was carried out. Patients with CC were identified using the criteria proposed by Levenson and Parisier. RESULTS: Thirty-five patients with CC were identified. In 30 (86%) patients, the diagnosis was made during ear examination and the remaining five (14%) patients were diagnosed during myringotomies. Preoperative CT scans were available in 17 patients. The findings on CT scans were classified into four categories based on the ossicular chain and mastoid septae status as well as the presence or absence of middle ear and mastoid opacification. Intraoperatively, 22 (63%) patients were found to have extensive cholesteatomas with or without extension beyond the mesotympanum. Eleven of these 22 patients had ossicular chain erosion and five were later found to have recidivism. Preoperative CT scan accurately predicted the extent of the cholesteatoma seen during surgery in 14/17 (82%) and ossicular chain status in 15/17 (88%), while micro-otoscopy predicted the extent of the existing pathology in only 10/35 (29%). Intraoperative location and size of CC influenced the type of surgical approach, status of ossicular chain, postoperative hearing level and rate of recidivism. CONCLUSIONS: Children still present with late stage CC. Micro-otoscopy is insufficient to clearly delineate the extension of CC. Preoperative CT scan is essential in defining the extent of existing pathology. The intraoperative CC size and location influence the outcome of surgery. Early surgical intervention and long-term follow-up are essential. PMID- 12633922 TI - Naive and memory T cells in hypertrophied adenoids in children according to age. AB - The anatomic location of the adenoid implies that this organ is the first site of contact with inhaled antigens. Depending on the expression of different isoforms of the CD45 molecules, T cells can be divided into naive (CD45RA(+)) and memory (CD45R0(+)) cells, the latter representing T cells that have already been exposed to antigens. The purpose of this study was to analyse the lymphoid cells' subsets in adenoids and relate the findings to the age. The analysed material was adenoid tissue removed on the grounds of hypertrophy from 22 children. The patients were divided into two groups: up to 5 and above 5 years of age. The analyses of the lymphocytes subpopulations in the adenoid were performed in an EPICX XL (Coulter) flow cytometry. The results are expressed as the percentage of positively labeled cells (CD4(+), CD8(+), CD4(+)/CDB(+), CD4(+)CD45RA(+), CD8(+)CD45RA(+), CD4(+)CD45R0(+), CD8(+)CD45R0(+)). The percentage of CD4(+)/CD45R0(+) in children up to 5 years of age was significantly lower than in older children. We found the positive regression between age and the percentage of CD4(+) cells was CD45R0(+) (r=0.64). There were no statistically significant differences between study subgroups for the other parameters. The positive regression for CD4(+)/CD45R0(+) cells and age may result from increased stimulation by bacterial, viral and other antigens. Our results indicate that the adenoid have an important role in the development of an immunological memory among younger children. PMID- 12633923 TI - Video-optic cable endoscopy forceps. AB - We present a video-endoscopic technology for foreign body extractions with forceps. This can easily be followed and is recordable on video monitor in excellent quality. Instead of a rigid rod lens telescope, a fiberoptic video endoscope is inserted into the optical channel of a bronchoscopic forceps. This keeps the endoscopic device lightweight and easily maneuverable in different angles. The principle can be applied to other rigid diagnostic and therapeutic endoscopic equipment and represents a potential alternative technology to the use of cameras attached to rigid telescopes. PMID- 12633924 TI - Speech perception of Mandarin Chinese speaking young children after cochlear implant use: effect of age at implantation. AB - OBJECTIVE: The purposes of this study were to examine (1). whether the benefit of cochlear implants in Mandarin-speaking congenitally deaf children persists over a 2-year follow-up period, and (2). whether the age of the child at implantation affects the acquisition of speech perception by young deafened children. METHOD: Sixteen congenitally deaf children received implants of Nucleus CI24M devices. Speech perception tests were conducted using the Mandarin Auditory Perception Test Battery at 6-month intervals after connection of the device. We determined the benefit of cochlear implants by monitoring percent correct scores between 12 and 24 months after implantation. Regression analysis was used to correlate the measures of progress and the age at implantation. RESULTS: In the comparison of performance at 12 and 24 months post-connection, subjects performed significantly better at 24 than at 12 months in spondee, vowel, and consonant tests (P<0.05). In phrase and sentence tests, the mean score improved over time with device use, but the difference was not significant (P=0.066 and 0.067, respectively). However, no obvious improvement in average score was found in tone tests (P=0.386). Progress (improvement in speech perception) at 12 and 24 months had moderate negative correlations with age at implantation in spondee, vowel, phrase and sentence tests (P<0.05), but no correlation with age in consonant (P=0.20) and tone tests (P=0.26). CONCLUSION: The result, in accordance with the reports on Western language-speaking children, showed cochlear implants increasingly benefit Mandarin-speaking congenitally deaf children over a 2-year post implantation period. Because age at implantation negatively correlates with improved speech perception after implantation, congenitally affected children with profound deafness (if they cannot obtain enough benefit from hearing aids) should receive implants as early as possible. PMID- 12633925 TI - Nasal endoscopy in the treatment of congenital lacrimal sac mucoceles. AB - PURPOSE: To determine the incidence of intranasal cysts associated with lacrimal sac mucoceles and the cure rate with nasal endoscopic cyst marsupialization. DESIGN: Interventional case series. METHODS: SETTING: University-affiliated teaching hospital. PATIENT POPULATION: Twenty-five infants with non infected or infected lacrimal sac mucoceles or dacrocystitis without obvious mucocele were consecutively enrolled. INTERVENTION PROCEDURES: Management included local lacrimal massage, parenteral antibiotics, and when still symptomatic, nasolacrimal duct probing with concomitant nasal endoscopy. Intranasal cysts identified were marsupialized until the distal end of the nasolacrimal duct probe was visualized. MAIN OUTCOME MEASURES: Presence of intranasal cyst identification and cure rate. RESULTS: Infants were 4 days to 10 weeks old (mean 19 days). Forty eight percent had a bluish cutaneous mass inferior and lateral to the lacrimal sac. Twenty percent were bilateral. At presentation, 76 percent had dacrocystitis. Fourteen percent had respiratory distress. Only one child responded to medical management. At endoscopy, 23 of 24 infants had ipsilateral intranasal cysts. The one child without nasal cyst had recurrent dacrocystitis and no mucocele. All children with mucocele were cured except one child with residual nasolacrimal duct obstruction. CONCLUSIONS: Lacrimal sac mucoceles were almost always associated with intranasal cysts. Nasal endoscopy is a valuable addition to the treatment plan for lacrimal sac mucoceles not responding to a brief trial of massage or infantile dacrocystitis. To avoid potential complications, we recommend against waiting until infection occurs before proceeding with surgery. PMID- 12633926 TI - Surgery of the lateral nasal wall and ethmoid: effects on sinonasal growth: an experimental study in rabbits. AB - OBJECTIVE: To assess the impact of lateral nasal wall surgery on sinonasal growth METHODS: Twenty young New Zealand White rabbits, 6 weeks of age, were included in this experimental study. Surgery was performed on two groups of ten animals each (series I and II). Entrance to the left nasal cavity is achieved through the nasal dorsum via mobilization and rotation of the left nasal bone. Series I: partial resection of the lateral nasal wall (including the ostium to the maxillary sinus) on the left side. Series II: partial resection of the lateral nasal wall and anterior ethmoid. Follow-up period was 20 weeks. Twenty rabbits served as controls. RESULTS: In series I, all skulls have grown normally. In series II the nasal dorsum has also developed symmetrically. Snout length and growth of upper jaw are normal; there is no malocclusion. Three skulls show a slight deviation of the nasal dorsum (two to the left, one to the right). Morphometric measurements of 20 points on the skulls show no significant difference between the control group and the experimental series I and II. CONCLUSION: This experimental study demonstrates that visually controlled partial resection of the bony sinonasal wall, with or without resection of the anterior ethmoid does not affect later development of nose and upper jaw on condition that eventually underlying cartilage is preserved. Contradictory results from other experimental studies, previously published and concerning negative effects of sinus surgery, might be attributed to surgical traumatization of intranasal cartilage structures, in particular, the upper lateral cartilages. PMID- 12633927 TI - Endoscopic treatment of congenital choanal atresia. AB - This report outlines the effectiveness of the transnasal endoscopic approach for the treatment of congenital choanal atresia. Fourteen patients with age between 5 days and 15 years were treated using this approach by means of 0 and 30 degrees 4 or 2.7 mm telescopes: three cases were bilateral while 11 were unilateral. The time of stenting ranged from 3 to 6 days (mean 4 days) in the three newborn patients and between 1 and 2 weeks (mean 1 week) in the other nine patients. In the last two cases, stenting was not carried out. The overall follow-up period ranged from 2 to 64 months (mean 31 months). Only one restenosis was observed in this series 2 months after surgery. This restenosis was successfully treated by endoscopic revision. The authors believe that a correct repositioning of mucosal flaps after the resection of the atretic plate is of paramount importance in avoiding restenosis of the neochoana and in reducing the time and/or the use of stenting. PMID- 12633928 TI - Acute mastoiditis in children: Pseudomonas aeruginosa as a leading pathogen. AB - OBJECTIVE: Acute mastoiditis is a serious bacterial infection of the temporal bone and is the most common complication of otitis media. The goal of this study is to assess the clinical features, pathogens, management, and outcome of acute mastoiditis in children in northern Israel. METHODS: A systematic review of medical records of all children who were admitted with acute mastoiditis from January 1990 through December 2000. RESULTS: Fifty-seven children were included. Median was age: 36 months. In 26 patients (45.6%) mastoiditis complicated the first episode of acute otitis media (AOM). Twenty-five children (44%) received antibiotic treatment prior to admission. Frequent symptoms included mastoid area erythema in 54 children (94.7%), proptosis of the auricle in 52 children (91.2%) and fever in 43 children (75.4%). Middle ear, and subperiostal culture yielded growth of pathogen in 30 children (75%), two cultures yielded more then one pathogen. The most frequent pathogens were: Pseudomonas aeruginosa in 10 children (25%), Streptococcus pneumoniae in eight children (20%), Group A streptococcus in six children (15%). The highest incidence of Streptococcus pneumoniae was found in children who did not suffer from AOM before admission (35 vs. 5%). Fifty-two (91.2%) children were cured with antibiotic treatment alone. Seventeen children underwent computed tomography (CT) of the mastoid. Mastoid bone destruction was demonstrated in six children and subperiostal abscess in eight. Mastoidectomy was performed in five children. CONCLUSIONS: The diagnosis of acute mastoiditis can be made on clinical basis alone requiring CT only when complications are suspected. Half of the children admitted with acute mastoiditis had no previous history of recurrent AOM. In those children S. pneumoniae was the leading pathogen while P. aeruginosa was more prevalent in children with recurrent AOM. Most of the children recovered with medical therapy alone, without surgical intervention. PMID- 12633929 TI - Congenital laryngeal cyst: one or two cysts. AB - The case presented is of a neonate with extreme respiratory distress immediately after birth, caused by a large laryngeal cyst. The vertical diameter of the cyst was larger than the height of the neonatal larynx. First excision of the cyst was performed in the region of the prominent aryepiglottic fold. Three weeks later, because of a recurrence of dyspnea, excision of the prominent wall of an obstructing cystic lesion was performed in the ventriculus Morgagni. PMID- 12633930 TI - Hamartoma of the middle ear: a case study. AB - Hamartomas and teratomas are rare tumors in children. Its location in head and neck is quite exceptional. An unusual case of hamartoma in the middle ear is reported in a 5-month-old child. The tumor was isolated without associated malformation. Definitive diagnosis was made after surgery and based on histological results. At 2 years follow-up, no recurrence was observed. CT-scan is useful in order to reach a final diagnosis. Although rare, this type of tumor should be considered in the differential diagnosis of a middle ear mass during childhood. PMID- 12633931 TI - Occult supraglottic lymphatic malformation presenting as obstructive sleep apnea. AB - Sleep disordered breathing and obstructive sleep apnea is commonly encountered in the pediatric population. In many cases, it is the result of oropharyngeal obstruction secondary to adenoidal or adenotonsillar hypertrophy. We describe an unusual case of a child with adenoidal hypertrophy who had an occult supraglottic lymphatic malformation that manifested as obstructive sleep apnea. The management of this lesion is discussed including the use of endoscopy, carbon-dioxide laser, and the decision to avoid a tracheotomy. Occult supraglottic lymphatic malformations (LMs) are a rare cause of obstructive sleep apnea, the diagnosis of which will be missed without fiberoptic laryngeal examination. They are challenging to manage because of the airway involvement and propensity to recur. PMID- 12633932 TI - Juvenile xanthogranuloma of the nasal cavity. AB - Juvenile xanthogranuloma (JXG) is the most common non-Langerhans histiocytosis, but it rarely occurs extracutaneously. It sometimes presents with associated neurofibromatosis and Juvenile chronic myalogenous leukemia. We present a case of nasal JXG and discuss the histological characteristics, the classification, and the management. PMID- 12633933 TI - Age-related changes in rat cerebral occludin and zonula occludens-1 (ZO-1). AB - The endothelial or epithelial tight junctions create a rate-limiting barrier to diffusion of solutes. A major determinant of the barrier function is the density of tight junction proteins. Since aging is associated with significant alterations in the blood-brain barrier (BBB) it is possible that specific tight junction proteins may be altered in the cerebrum of aging rats. To test this hypothesis, Western and Northern blot analysis were carried out to measure the steady-state level of occludin and zonula occludens-one (ZO-1) proteins and their mRNA in cerebral tissue of 3-, 12- and 24-month-old rats. The cerebral occludin content in 24-month-old rats (732.5+/-99.9 arbitrary units) was significantly reduced compared to 12-month-old rats (1043.4+/-131.8) or 3-month-old rats (1021.4+/-62.8), P<0.01. The cerebral ZO-1 protein content in 24-month-old rats (161.7+/-8.1 arbitrary units) and 12-month-old rats (144.3+/-35.9) were not significantly reduced compared to 4-month-old rats (189.0+/-27.2). The occludin mRNA content relative to G3PDH mRNA was 1.11+/-0.05, 1.11+/-0.07 and 1.00+/-0.05 in 3-, 12- and 24-month-old rats, respectively. The differences did not achieve statistical significance. The ZO-1 mRNA content of cerebral tissue relative to G3PDH mRNA was significantly increased in 24-month-old rats compared to 3-month old rats (1.280+/-0.030 vs. 0.956+/-0.038), P<0.001. It is concluded that aging in rats may alter the molecular anatomy of the BBB by altering the content of select structural proteins of tight junctions. PMID- 12633934 TI - B cell precursors in senescent mice exhibit decreased recruitment into proliferative compartments and altered expression of Bcl-2 family members. AB - In vitro, aged pro-B cells generally exhibit limited expansion in response to IL 7 when compared to young pro-B cells. CFSE-labeling in vitro indicated that aged mice have a lower frequency of pro-B cells which are capable of undergoing extensive proliferation upon stimulation with exogenous IL-7. Protein levels of the survival molecule Bcl-x(L) were consistently reduced in IL-7 expanded aged pro-B cells. Levels of both Bcl-2 and Baxalpha proteins were variable in aged B cell precursors. The expansion of aged pro-B cells in response to IL-7 in vitro correlated inversely with the ratio of the pro-apoptotic protein Baxalpha to the survival protein Bcl-2. Expansion of aged pro-B cells in vitro is likely dictated by both recruitment of pro-B cells into proliferative compartments and their survival; in aged B cell precursors, the latter is favored by low Baxalpha to Bcl 2 protein ratios. PMID- 12633935 TI - Germinal center B cells in Peyer's patches of aged mice exhibit a normal activation phenotype and highly mutated IgM genes. AB - Systemic and mucosal humoral immune responses have been reported to exhibit an age related decline. However, differences in primary and memory responses at mucosal sites in aged mice have been noted. In an effort to begin characterizing deficiencies in the mucosal system of aged mice, we examined the B cell compartment of gut associated Peyer's patch lymphoid tissue. To our surprise, we found that germinal center (GC) B cells from aged B6D2F1 mice (24-26 months) were present at similar frequencies and exhibited a normal activation phenotype such as upregulation of B7.1, B7.2 and CD44, and downregulation of CD23, CD62L and CD38 as that observed in younger mice (2.5-4 months). As expected, Peyer's patch GC B cells from aged mice expressing V(H)X24 genes displayed higher somatic mutation frequencies compared with younger mice. However, this was particularly striking in IgM sequences where high mutational loads suggested we were sampling memory cells. It is conceivable that B-cells expressing these genes reflect the presence of a mucosal memory compartment in aged mice that either retains flexibility in effector function or is committed to the secretion of IgM antibody. PMID- 12633936 TI - Characterisation of the interaction between WRN, the helicase/exonuclease defective in progeroid Werner's syndrome, and an essential replication factor, PCNA. AB - Ageing is linked to the accumulation of replicatively senescent cells. The best model system to date for studying human cellular ageing is the progeroid Werner's syndrome (WS), caused by a defect in WRN, a recQ-like helicase that also possesses exonuclease activity. In this paper, we characterise the interaction between WRN and an essential replication factor, PCNA. We show that wild-type WRN protein physically associates with PCNA at physiological protein concentrations in normal cells, while no association is seen in cells from patients with WS. We demonstrate co-localisation of WRN and PCNA at replication factories, show that PCNA binds to two distinct functional sites on WRN, and suggest a mechanism by which association between WRN and PCNA may be regulated in cells on DNA damage and during DNA replication. PMID- 12633937 TI - Altered RNA structural constituents in aging and vitamin E deficiency. AB - Ribonucleoprotein (RNP) containing structural constituents in hepatocyte nuclei of adult, old and adult, vitamin E-deficient rats were investigated to assess the effect of aging and increased oxidative stress on nuclear functions. Fibrillar centres (FCs), dense fibrillar (DFC) and granular (GC) components of nucleoli as well as perichromatin granules (PGs) in the nucleoplasm were preferentially evidenced by the ethylenediaminetetracetic acid (EDTA) method and measured by computer-assisted morphometric procedures. FCs size and the percentage of nucleolar surface occupied by FCs significantly decreased during aging and vitamin E-deficiency. The percentage of nucleolar surface occupied by GC and DFC remained unchanged in adult and old rats, but in vitamin E-deficient animals GC increased and DFC decreased significantly. PG density significantly changed in aging and vitamin E-deficiency. Functionally, FCs, DFC and GC constitute sites of transcription and processing of ribosomal RNA while PGs are involved in intranuclear storage and transport of messenger RNA. Thus, the present structural changes during aging and vitamin E-deficiency correlate with a decay of nuclear responsiveness to cellular metabolic needs. Considering the antioxidant action of alpha-tocopherol, our data lend further support to the importance of free radical production and control in the aging process. PMID- 12633938 TI - Physical activity prevented age-related decline in energy metabolism in genetically obese and diabetic rats, but not in control rats. AB - Laboratory rats are normally confined to cages that markedly restrict their physical activity. In these rats, the resting energy expenditure accounts for 90% of the total daily energy expenditure, while the daily physical activity in humans consumes 30% of the total daily energy expenditure. Otsuka Long Evans Tokushima Fatty (OLETF) rats have been developed as a model of non-insulin dependent diabetes mellitus (NIDDM) with mild obesity, and obesity is an important factor that induces diabetes in this strain. We implemented a running wheel exercise regimen that was the equivalent of normal physical activity to provide light exercise for OLETF rats. The purpose of the study was to determine if light exercise improves the age-related decline in energy metabolism and glucose intolerance in OLETF rats. The effects were also compared in control Long Evans Tokushima (LETO) rats. From 12 to 46 weeks of age, the rats performed a running-wheel exercise (3000 m/day). Energy metabolism was determined at 8-week intervals. The typical increase in body weight was significantly decreased in OLETF rats in response to exercise, while no significant effect was observed in LETO rats. Energy expenditure and basal metabolic rate (BMR) per kilogram body weight (not whole-body weight) were increased by exercise in OLETF rats, but not in LETO rats. At 46 weeks of age, after exercise, the blood glucose and hemoglobin (Hb)A1c levels, as well as the plasma levels of insulin, triglyceride, cholesterol, and leptin significantly decreased in OLETF rats, while only the plasma levels of cholesterol and leptin significantly decreased in LETO rats. Light exercise thus appears to be beneficial for preventing age-related decline in energy metabolism and glucose intolerance in OLETF rats. PMID- 12633939 TI - Oxidative inactivation of CD45 protein tyrosine phosphatase may contribute to T lymphocyte dysfunction in the elderly. AB - The decline in T lymphocyte function during ageing has been linked to changes in intracellular signalling pathways. Oxidative damage has long been thought to be involved in the ageing process and we investigated a link between ageing, oxidation and T cell signalling focusing on the membrane phosphatase CD45. We investigated the relative sensitivity of CD45 to oxidative inactivation and then compared the phosphatase activity of CD45 in blood lymphocytes from elderly and young volunteers and related this to intracellular levels of the antioxidant glutathione. The CD45 phosphatase was particularly sensitive to oxidative inactivation compared to total Jurkat T cell PTP activity. The IC50 with H(2)O(2) was 3 mM for CD45 at which concentration there was a minimal decrease in global PTP activity. In normal peripheral blood CD4(+) T cells the IC50 was much lower at 54 microM. In a group of elderly healthy individuals, whose T cell responses to mitogen were depressed, PB lymphocyte CD45 phosphatase activity was decreased by about 60% compared to young controls. There was no difference in intracellular levels of glutathione. The loss of CD45 activity in lymphocytes from the elderly may underlie poor T cell function associated with ageing. The relative sensitivity of CD45 to oxidative damage may result from its location in the plasma membrane, where it might be more accessible to extracellular oxidants. PMID- 12633940 TI - Study of age-association with cytokine gene polymorphisms in an aged Irish population. AB - The release of cytokines is of crucial importance in the regulation of the type and magnitude of the immune response in the elderly. A number of studies have shown different levels of cytokine production in the elderly. In the present study, a range of polymorphisms were chosen within the genes of cytokines (IL-2, IL-6, IL-8, IL-10, IL-12 and IFN-gamma) that have been observed at different levels within the elderly and analysed for age-association. No association was observed for the polymorphic cytokine markers and the healthy aged Irish population (or with respect to gender) examined in this study. These findings would suggest that polymorphism of cytokine genes may not play as crucial a role in healthy ageing as previously believed. PMID- 12633941 TI - Role of stromal cells and their products in protecting young and aged B-lineage precursors from dexamethasone-induced apoptosis. AB - Bone marrow stromal cells are potent providers of stimuli that induce proliferation of B-cell precursors. We proposed that stromal cells play a role in protecting B-lineage cells from corticosteroid-induced apoptosis. We found that stromal cells protected B-cell precursors from dexamethasone-induced apoptosis, but this did not strictly correlate with interleukin-7 (IL-7) production. To determine if stromal-derived factors were involved in protection of B-cell precursors from apoptosis, we examined the activity of three lymphopoietic growth factors: IL-7, stem cell factor (SCF), and insulin-like growth factor-1 (IGF-1). Either IL-7 or IGF-1 alone protected B-cell precursors from dexamethasone-induced apoptosis. The combined activities of IGF-1 and IL-7 were additive rather than synergistic. SCF did not protect B-cell precursors from apoptosis. Aging altered the ability of B-cell precursors to respond to protective stimuli induced by IL-7 and IGF-1. Precursors from aged animals were deficient in ability to modulate expression of apoptosis regulatory genes Bax, Bcl-2, and Bcl-x in comparison to B cell precursors from young animals. Taken together, these results suggest that stromal cells can protect B-lineage precursors from a corticosteroid-induced apoptotic signal, protection is mediated by stromal-derived cytokines, and aging decreases the ability of B-cell precursors to respond efficiently to protective stimuli. PMID- 12633942 TI - A strategy for the ubiquitous overexpression of human catalase and CuZn superoxide dismutase genes in transgenic mice. AB - In the present study, we generated transgenic mice that overexpress catalase or CuZn superoxide dismutase (CuZnSOD) in all tissues using large genomic DNA fragments. An 80 kb human genomic DNA, containing the 33 kb human CAT gene as well as the 41 kb of 5' and the 6 kb of 3' flanking regions, was obtained by screening a human P1 library and was used to produce transgenic mice Tg(CAT). Transgenic mice Tg(SOD1) were produced by a similar strategy using a 64 kb human genomic DNA containing the 10 kb human SOD1 gene and the 27 kb of both 5' and 3' flanking regions. Catalase mRNA levels were 2-6- fold higher and catalase activity levels were 2-4- fold higher in the various tissues of the hemizygous Tg(CAT) mice compared with wild type mice. The mRNA levels for CuZnSOD were 2-12- fold higher and the CuZnSOD activity levels were 2-5- fold higher in the hemizygous Tg(SOD1) mice compared with wild type mice. In summary, our study demonstrates that a strategy of using large genomic DNA containing either the entire human CAT or SOD1 gene with large flanking regions gives ubiquitous increased expression of CuZnSOD and catalase. In addition, the expression of catalase closely reflects the tissue specific pattern found in the endogenous gene. These transgenic mice will be useful in studying the role of oxidative stress/damage in aging and age-related pathologies. PMID- 12633943 TI - Ageing exacerbates the cardiotoxicity of hydrogen peroxide through the Fenton reaction in rats. AB - Reactive oxygen species (ROS) are involved in the post-ischemic reperfusion syndrome of the myocardium. Moreover, ageing is associated with an increased cardiac sensitivity to both ischemia and reperfusion. The aim of the present study was to determine whether the lower tolerance of aged hearts to reperfusion could be due to an increased sensitivity to the ROS that are produced during the early phase of reperfusion. For this purpose isolated perfused hearts from adult (4 months) and aged (24 months) rats were perfused with a buffer containing 150 microM of hydrogen peroxide (H(2)O(2)) in presence or absence of deferoxamine mesylate (150 microM), an iron chelator. H(2)O(2) perfusion was continued until left ventricular developed pressure had decreased up to 20% of its initial value. Ageing led to a significant reduction of the duration of the H(2)O(2) perfusion required for inducing a 80% functional alteration. Although deferoxamine did not affect this parameter in adult rats, it significantly increased the duration of H(2)O(2)-perfusion in senescent hearts (control: 14.0+/-0.9 min vs. deferoxamine: 18.1+/-1.0, P<0.05). Similarly, ageing aggravated cardiac contracture induced by H(2)O(2)-perfusion. Again, deferoxamine, which had no effect on this parameter in young adult hearts, significantly reduced the contracture of senescent rat hearts. To conclude, our data clearly show that ageing is associated with an increased sensitivity of the myocardium to hydrogen peroxide, which is partly reversed by iron chelation. These results suggest that the iron-catalyzed Fenton reaction producing hydroxyl radicals might be greater in hearts from senescent rats than in hearts from young adults. PMID- 12633944 TI - In vitro aging of macaque adherent cells: similar pattern of cellular aging between human and macaque. AB - To explore new models for human cellular aging as well as to evaluate aging of the macaques, profiles of cellular aging in macaques were studied. Adherent cells were obtained from five Japanese macaques (Macaca fuscata), 14 long-tailed macaques (Macaca fascicularis), two bonnet monkeys (Macaca radiata) and a rhesus monkey (Macaca mulatta). A total of 35 cultures were performed and cell morphology, doubling time, telomere length and telomerase activity were studied. They were classified into three groups; group I: cell strains with a definite replicative life-span (-41 PDLs) (presence of M1), group II: cell strains with a limited extension of replicative life-span (79-106 PDLs) with p53 mutation(s) (presence of M2), and group III: a cell strain with an indefinite replicative life-span (>150 PDLs) with characteristics of transformation. Except for the last group, telomerase activity was not observed. Macaque cells demonstrated three chronological patterns comprising both human and rodent patterns, however, presence of the two limits of proliferation in vitro grants macaque cells to be more appropriate than rodents in both studying human aging and oncogenesis. PMID- 12633946 TI - 30th anniversary year of Surgical Neurology. PMID- 12633948 TI - The effect of short-term antifibrinolytic therapy on experimental vasospasm. AB - BACKGROUND: Antifibrinolytic therapy is effective in preventing rebleeding in cases of aneurysmal subarachnoid hemorrhage (SAH). The major disadvantage of this therapy is the increase in ischemic complications, which is supposed to be due to cerebral vasospasm. In this study the effect of short-term antifibrinolytic therapy on arterial vessel narrowing after SAH was investigated utilizing the rat femoral artery vasospasm model. METHODS: Twenty-four rats were divided into four groups of six animals each. Autologous blood (0.1 mL) was applied to the 1-cm segment of right femoral artery wrapped with a silicone cuff. In Group 1 the animals did not receive any treatment. In Groups 2, 3, and 4 150 mg/kg tranexamic acid (AMCA) was given orally for 3, 5, or 7 days respectively, starting from postoperative day 1. A 1 cm segment of each femoral artery was harvested on the 8(th) postoperative day. Morphologic analyses were performed using the parameters, radial wall thickness and cross luminal area under the light microscope. In addition, two samples from each group were examined by transmission electron microscope (TEM) to confirm the morphologic changes. RESULTS: There was a gradual decrease in cross luminal area and gradual increase in vessel wall thickness directly proportional with time. However, the vasospastic changes that occurred in Group 2 (received AMCA for 3 days) were not significantly different from those of Group 1 (nontreated). CONCLUSION: It was concluded that antifibrinolytic treatment for the first 3 days may prove useful in cases of clinical aneurysmal SAH. However, if this treatment is used for more than 3 days, arterial vessel narrowing is significantly increased. PMID- 12633950 TI - Vertebrobasilar artery dissection presenting with simultaneous subarachnoid hemorrhage and brain stem infarction: case report. AB - BACKGROUND: Intracranial dissecting aneurysms have been associated with subarachnoid hemorrhage (SAH) or cerebral ischemia. We encountered a patient presenting with simultaneous subarachnoid hemorrhage and brainstem infarction caused by a dissecting aneurysm of the vertebrobasilar artery, which was diagnosed by magnetic resonance imaging (MRI) but did not show abnormal findings on cerebral angiography. CASE DESCRIPTION: A 55-year-old man had sudden onset of headache and left abducens palsy. Computed tomography revealed a subarachnoid hemorrhage localized in the left prepontine cistern and the left cerebellomedullary fissure. Cerebral angiography showed neither a saccular aneurysm nor fusiform dilatation causing the subarachnoid hemorrhage. MRI demonstrated a small infarction in the left dorsal pons, and an intramural hematoma of the left vertebral artery and lower basilar artery. CONCLUSION: This is a rare case of a vertebrobasilar dissecting aneurysm that simultaneously caused both SAH and brain stem infarction. MRI should be performed in the acute phase of SAH of unknown origin to determine the possible coexistence of a dissecting aneurysm, as occurred in this case. PMID- 12633951 TI - Chronic intraventricular encapsulated hematoma presenting unique radiological features: a case report. AB - BACKGROUND: Chronic encapsulated hematoma is relatively rare and the exact mechanisms of this condition remain uncertain. In this report, we present a case of chronic encapsulated hematoma in the lateral ventricle and discuss the pathophysiology of this condition. CASE DESCRIPTION: A 68-year-old-woman presented with progressive gait disturbance. Magnetic resonance imaging (MRI) showed a dumbbell-shaped mass arising from the wall of the right lateral ventricle. The mass connecting to the ventricle wall mimicked a hemorrhagic tumor and the other part mimicked a cystic mass. Cerebral angiography demonstrated arteriovenous shunt in the right lateral ventricle wall. The operative findings revealed that the part arising from the ventricle wall contained hematoma showing various stages of coagulation and that the cystic mass visible on MRI was a granulomatous solid mass. Histologic examination revealed that the capsules of both were compatible with chronic encapsulated intracerebral hematomas. CONCLUSION: In the present case, one part of the mass contained only granulomatous tissue, indicating the possibility that the mass was undergoing spontaneous resolution. This fact might indicate that the presence of neo vascularization in the capsule is not the sole factor in the expansion of hematoma. PMID- 12633952 TI - Title comparison of primary and secondary spinal epidural abscesses: a retrospective analysis of 29 cases. AB - BACKGROUND: Spinal epidural abscesses (SEA) are uncommon. In certain regions their incidence is rising. Vague initial presentation may result in delayed diagnosis. Familiarity with SEA is imperative because if not treated expeditiously, they can have devastating neurologic sequelae. METHODS: A retrospective analysis using analysis of variance (ANOVA) was performed on patients diagnosed with SEA between 1980 through 2000. The patients were assigned to one of the two defined groups: primary (PSEA) consisted of SEA alone; secondary (SSEA) included SEAs with vertebral osteomyelitis. Both groups were compared for factors including survival, age, diagnostic accuracy, etiology, hospitalization, management, and outcome. RESULTS: Twenty-nine cases were identified. Four (23.7%) were PSEAs and 25 (86.2%) were SSEAs. Mean age (52.7 vs. 53.2 years) and mean duration of hospitalization (39.2 vs. 38.6 days) were comparable in both groups (p = 0.9). The admitting diagnosis was correct in 75% of PSEA and 20% of SSEA cases (p = 0.1). Staphylococcus aureus was present in 75% and 68%, respectively. In SSEA cases, 24% (n = 6) of the infections were consequent to spinal surgery. PSEA did not show a predilection for any level. Most (56%) cases of SSEA occurred at the lumbar levels. The entire PSEA group and 58.3% of the SSEA group underwent surgery (p = 0.06), 75% versus 40.9% had a good outcome for PSEA and SSEA, respectively. CONCLUSIONS: PSEAs are very rare. Both groups have similar characteristics. Staphylococci remain the predominant etiologic agent. PSEAs are treated by surgery; SSEAs are managed surgically or conservatively. PMID- 12633954 TI - A recurrent intradural cervical neurenteric cyst operated on using an anterior approach: a case report. AB - The neurenteric cyst is an uncommon congenital lesion. In most reported cases, it has been operated on via a posterior approach using a laminectomy, despite the fact that the cyst is usually located ventral to the spinal cord. Reports have shown that early postoperative results have been good with the posterior approach, but very few studies of the long-term postoperative recurrence of neurenteric cysts have been conducted. Here, we report on a case of recurrent neurenteric cyst that was operated on using an anterior approach.A 42-year-old woman presented with a cervical neurenteric cyst that had recurred eight years after its partial removal via a posterior approach. The patient complained of pain on the lateral side of her upper arms, and an magnetic resonance imaging showed that the recurrent cyst was located ventral to the spinal cord and compressed the cord dorsally at the C4-6 level. The patient was operated on via an anterior approach using a vertebrotomy at the lower half of C5 and the upper half of C6. The cyst was attached to the spinal cord firmly and was subtotally removed, with the thickest portion adhering to the cord not being removed. The caudal end of the cyst was observed with the assistance of a rigid endoscope.A neurenteric cyst may recur after partial removal, and the patient's condition may deteriorate during postoperative follow-up. The anterior surgical approach provides good visualization and facilitates safe removal of the lesion. PMID- 12633956 TI - Combining MEG and MRI with neuronavigation for treatment of an epileptiform spike focus in the precentral region: a technical case report. AB - BACKGROUND: Epileptic foci are often located in the vicinity but not necessarily within the boundaries of intra-axial brain tumors. Resection of these tumors is based on two major goals: first, maximizing tumor removal without provoking new neurologic deficits, and second, minimizing epileptic seizure activity. Magnetic source imaging (MSI) depicts the generators of magnetic fields overlaid on individual magnetic resonance (MR) images. Established application areas are lesions located adjacent to or partly within the sensory and motor cortex, or in the depth of the brain, necessitating a surgical approach through functionally highly relevant cortical regions. Magnetoencephalography (MEG) is also applicable for epileptiform spike foci recording during interictal activity. CASE DESCRIPTION: A patient with a recurrent glioma close to the Rolandic cortex scheduled for epilepsy and tumor surgery was investigated with MSI. The MSI data showed an epileptiform spike focus outside the tumor boundaries. The resulting MSI images were integrated into our neuronavigation system. This procedure allowed for the preoperative identification of the sensory and motor cortex, the precise localization of the epileptiform spike focus, and careful planning of the surgical procedure. In this case, we were able to safely resect the recurrent tumor and the epileptiform spike focus under general anesthesia using MSI-based neuronavigational guidance but no conventional intraoperative mapping techniques. CONCLUSION: Magnetic source imaging can be a valuable, noninvasive method for planning and performing tumor resections in high-risk brain regions, especially if an epileptiform spike focus has to be localized and included into the resection strategy. PMID- 12633961 TI - Meningeal hemangiopericytomas: long-term outcome and biological behavior. AB - BACKGROUND: The authors present a retrospective clinical analysis of meningeal hemangiopericytomas. The long-term outcome and biologic behavior, including local recurrence and extraneural distant metastasis, are elucidated. METHOD: Clinical records and radiologic findings of 31 cases with meningeal hemangiopericytoma treated at Seoul National University Hospital and Asian Medical Center between 1982 and 1999 were carefully reviewed. The final outcome was determined by direct or phone contact and questionnaire by mail. The duration of follow-up was from 1 to 216 months (mean 77). All patients underwent craniotomy and Simpson Grade I or II resection was possible in 24 patients. Conventional radiotherapy was given in 11 patients (in 5 after total excision and in 6 for residual mass) and stereotactic radiosurgery was conducted in 6 cases. RESULTS: Intracranial recurrence was seen in 12 cases (38.7%) and the mean period before the first recurrence was 104 months. The recurrence was at the primary site in 11 cases and diffuse leptomeningeal seeding occurred in the remaining case. The 5-year recurrence free rate was 59.2% and the extent of excision was the significant factor (72.7% in total excision group and 20.8% in the incomplete excision group, p = 0.006). In four patients (12.9%), extraneural metastases developed at an average of 107 months. Six patients died during the follow-up period; however, 2 of these died of unrelated causes. CONCLUSIONS: Complete excision is the most important factor in reducing recurrence. However, recurrence may occur even after complete excision. Careful long-term follow-up is mandatory because of the long disease-free interval. PMID- 12633964 TI - Intracranial chloroma in hypereosinophilic myelofibrosis. AB - BACKGROUND: Chloroma, also called granulocytic sarcoma, is a localized extramedullary tumor composed of leukemic myeloid cells. It is a rare tumor that can occur in various locations. The association of chloroma with leukemic disease or myeloproliferative disorders is limited to isolated case reports. CASE DESCRIPTION: We report a case of intracranial chloroma in an elderly man with myelofibrosis and progressive hypereosinophilia. The presence of leukemic cells within his pleural aspirate suggested an incipient acute leukemic state. CONCLUSION: We report the first case of intracranial chloroma associated with hypereosinophilia developing in the course of myelofibrosis. The significance of hypereosinophilia in predicting the likelihood of development of central nervous system chloroma and acute leukemia in a patient with myelofibrosis needs further evaluation. PMID- 12633968 TI - Percutaneous trigeminal ganglion balloon compression for treatment of trigeminal neuralgia--part I: pressure recordings. AB - BACKGROUND: The purpose of this study was to establish standards for the pressure monitoring system and to define the pressure pattern during percutaneous trigeminal ganglion compression for treatment of trigeminal neuralgia. METHODS: Seventy-five patients with intractable trigeminal neuralgia who underwent percutaneous trigeminal ganglion balloon compression were included in this study. A computerized pressure system was used for pressure monitoring and analysis. RESULTS: The procedural pressure patterns of the balloon opening pressure and the initial compression pressure were identified. On average, the balloon opening pressure was 2956 +/- 185 mm Hg in Meckel's cave (area 2) and it was much higher than that outside the foramen ovale (area 1, 2402 +/- 172 mm g), or in the posterior fossa (area 3, 2120 +/- 127 mm Hg) (p < 0.05). The average initial compression pressure in area 2 was 1204 +/- 105 mm Hg, and it was also significantly higher than those in area 1 (728 +/- 42 mm Hg) and area 3 (458 +/- 72 mm Hg) (p < 0.05). CONCLUSIONS: The pressure monitoring system has proven to be accurate, reliable, and extremely useful for monitoring the percutaneous trigeminal ganglion balloon compression procedure. PMID- 12633965 TI - Successful aqueductal plasty and stenting for tectal plate tumor after failed third ventriculostomy: a case report. AB - BACKGROUND: Tectal region tumors can lead to hydrocephalus secondary to aqueductal compression. Surgical options for these patients include extracranial cerebrospinal fluid (CSF) shunts, third ventriculostomy, and/or aqueductal plasty. In cases of third ventriculostomy failure, the accepted alternative is an extracranial CSF shunt. We report a patient in whom a repeat third ventriculostomy with aqueductal plasty and stenting was successful after a failed initial third ventriculostomy. CASE PRESENTATION: A 12-year-old patient with hydrocephalus secondary to a tectal tumor presented with headaches and blurry vision. She had no focal neurologic findings. She underwent a third ventriculostomy. Five months after the procedure she had recurrence of her symptoms. Therefore, she underwent a secondary third ventriculostomy with aqueductal plasty and stenting. She has been symptom-free for 1 year. CONCLUSION: Aqueductal plasty with stenting may be an alternative to CSF shunts in some patients with hydrocephalus because of aqueductal compression resulting from tectal tumors. PMID- 12633971 TI - Interhemispheric arachnoid cyst in the elderly: case report and review of the literature. AB - BACKGROUND: Preoperative differential diagnosis of interhemispheric cysts is sometimes difficult. CASE DESCRIPTION: We recently experienced a case of symptomatic interhemispheric arachnoid cyst in a 62-year-old woman. We reviewed interhemispheric arachnoid cysts in the elderly and the management of symptomatic interhemispheric arachnoid cysts in elderly patients. Symptomatic interhemispheric arachnoid cysts in the elderly are predominantly located on the right side, have a long history of progressive symptomology, occur predominantly in females, and have no communication with the subarachnoid space. Interhemispheric arachnoid cysts are usually not associated with agenesis of the corpus callosum in elderly patients, whereas interhemispheric nonarachnoid cysts are usually associated with agenesis of the corpus callosum, which will be clearly demonstrated on magnetic resonance imaging. CONCLUSIONS: It is highly possible that an interhemispheric cyst without agenesis of the corpus callosum in an adult is an arachnoid cyst. PMID- 12633972 TI - Practice patterns in the U.S. PMID- 12633976 TI - Research. PMID- 12633977 TI - The relationship of age to prepulse inhibition and habituation of the acoustic startle response. AB - Prepulse inhibition (PPI) of the startle response reflects an early stage of information processing that is abnormal in schizophrenia and certain other specific neuropsychiatric disorders that are distinguished by the inability to inhibit redundant or relatively irrelevant sensory, cognitive, or motor information. The goal of the present study was to characterize the effect of normal aging on PPI and habituation of the startle response and to examine the hypothesis that normal aging is characterized by a global decline in inhibitory function. Ninety-seven non-psychiatric controls (age range 18-88) were tested for startle eyeblink response using electromyogram (EMG) recording. Startle magnitude decreased and startle latency increased with aging. PPI demonstrated an inverted U-shaped function with age (greatest PPI at intermediate ages) while there was no significant effect of age on startle habituation. The results do not support the theory that aging is associated with a general decline in inhibitory function and contrast with previous studies that have compared only extreme age groups and have found no effects of age on PPI. PMID- 12633978 TI - Signs of REM sleep dependent enhancement of implicit face memory: a repetition priming study. AB - Faces are processed and stored in distinct neuroanatomical systems. Based on evidence of a critical role of sleep in memory processes, we investigated the impact of nocturnal sleep on implicit memories for faces in healthy men. Face repetition effects in reaction times were compared across sleep periods early in the night, which are dominated by slow wave sleep (SWS), and late in the night, where rapid eye movement (REM) sleep prevails, as well as across corresponding nocturnal intervals of wakefulness. An inverse priming effect was found selectively across REM sleep rich late sleep, as indicated by distinctly prolonged response latencies to previously presented faces compared with novel faces after this period of sleep (P<0.05). We assumed this inverse priming to reflect a facilitated identification of previously presented faces after extended REM sleep periods, thereby producing interference with the response generation in our task which did not require face identification but rather required recognizing formal features of the faces. This interpretation was supported by a supplementary experiment where enhanced positive repetition priming was found across late, REM sleep dominated sleep in a task requiring face identification. Together, these findings indicate that implicit face memories particularly benefit from REM sleep associated brain mechanisms. PMID- 12633980 TI - Mitochondrial and endoplasmic reticulum stress-induced apoptotic pathways are activated by 5-aminolevulinic acid-based photodynamic therapy in HL60 leukemia cells. AB - We studied the mechanism of the cytotoxic effects of 5-aminolevulinic acid-based photodynamic therapy (ALA-PDT; induction with 1 mM ALA for 4 h followed by a blue light dose of 18 J/cm(2)) on the human promyelocytic leukemia cell line HL60 using biochemical and electron microscopy methods. The disruption of mitochondrial membrane potential, deltapsi(m), was paralleled by a decrease in ATP level, unmasking of the mitochondrial antigen 7A6, release of cytochrome c into the cytoplasm, activation of caspases 9 and 3 and cleavage of poly(ADP ribose) polymerase (PARP). This was followed by DNA fragmentation. These data suggest that ALA-PDT activates the mitochondrial apoptotic pathway. The level of endoplasmic reticulum Ca(2+)-binding chaperones ERp57 and ERp72 and of anti apoptotic proteins Bcl-2 and Bcl-x(L) was decreased whereas that of Ca(2+) binding protein calmodulin and the stress protein HSP60 was elevated following ALA-PDT. Inhibition of the initiator caspase 9, execution caspase 3 and Ca(2+) dependent protease m-calpain, did not prevent DNA fragmentation. We conclude that, in our in vitro model, ALA-based photodynamic treatment initiates several signaling processes in HL60 cells that lead to rapidly progressing apoptosis, which is followed by slow necrosis. Two apoptotic processes proceed in parallel, one representing the mitochondrial pathway, the other involving disruption of calcium homeostasis and activation of the endoplasmic reticulum stress-mediated pathway. PMID- 12633981 TI - Preferential inactivation of paediatric solid tumour cells by sequential exposure to Merocyanine 540-mediated photodynamic therapy and Edelfosine: implications for the ex vivo purging of autologous haematopoietic stem cell grafts. AB - Paediatric solid tumours exhibit steep dose-response curves to alkylating agents and are therefore considered candidates for high-dose chemotherapy and autologous stem cell support. There is growing evidence that autologous stem cell grafts from patients with solid tumours are frequently contaminated with live tumour cells. The objective of this study was to perform, in a preclinical purging model, an initial assessment of the safety and efficacy of a two-step purging procedure that combined Merocyanine 540-mediated photodynamic therapy (MC540-PDT) with a brief exposure to the alkyl-lysophospholipid, Edelfosine. Human and murine bone marrow cells and Neuro-2a murine neuroblastoma, SK-N-SH human neuroblastoma, SK-ES-1 and U-2 OS human osteosarcoma, G-401 and SK-NEP-1 human Wilms' tumour, and A-204 human rhabdomyosarcoma cells were exposed to a fixed dose of MC540-PDT followed by a brief incubation with graded concentrations of Edelfosine. Survival was subsequently assessed by in vitro clonal assay or, in the case of CD34 positive haematopoietic stem cells, by an immunohistochemical method. Combination purging with MC540-PDT and Edelfosine depleted all tumour cells by >4 log while preserving at least 15% of murine granulocyte/macrophage progenitors (CFU-GM), 34% of human CFU-GM, and 31% of human CD34-positive cells. The data suggest that combination purging with MC540-PDT and Edelfosine may be useful for the ex vivo purging of autologous stem cell grafts from patients with paediatric solid tumours. PMID- 12633982 TI - Yellowing and IR-changes of spruce wood as result of UV-irradiation. AB - The yellowing and IR-changes of spruce wood as a result of UV-irradiation were studied using two different types of xenon lamps (lambda>300 nm; I(o)=50 mW cm( 2) and lambda>280 nm; I(o)=17.5 mW cm(-2)). Changes in the IR spectra as well as the yellowing of the irradiated wood surfaces show the influence of UV light on the wood modules. The UV-irradiation (72 h; lambda>300 nm; I(o)=50 mW cm(-2)) decreased the lignin content on the surface by up to 20% of the original values. The colour difference of yellowing (deltaE) exhibited a systematic trend to higher values with increasing irradiation time. Our results show that the photoyellowing (UV-Vis detection) correlates very well with lignin degradation (IR detection). This result is in agreement with the quinone formation as the chromophoric reaction product of lignin decay. The degradation, yellowing, and oxidation kinetics differed only little using different light sources. The absorbed light intensity, which depends on wavelength, the intensity distribution of the light source and the absorption spectrum of lignin, influenced the degradation rate. Under the current experimental conditions, the absorption spectrum of lignin was the most important factor. Therefore, irradiation with lambda>280 nm is useful for rapidly monitoring the UV-degradation of wood PMID- 12633983 TI - Cellular uptake, localization and photodynamic effects of haematoporphyrin derivative in human glioma and squamous carcinoma cell lines. AB - Uptake, intracellular concentration, localization and photodynamic effects of a haematoporphyrin derivative (HpD, Photosan-3) were compared in human glioma (BMG 1, wild-type p53) and squamous carcinoma (4451, mutated p53) cell lines. Concentration and time dependence of cellular uptake of HpD was assayed from methanol extracts and whole cell suspension spectroscopy, while localization was studied by fluorescence microscopy-based image analysis. Colony-forming ability, apoptosis, cell-cycle progression and cytogenetic damage (micronuclei formation) were investigated as parameters of photodynamic response following irradiation with red light. BMG-1 cells were more sensitive to the photodynamic treatment than 4451 cells, although the 4451 cells accumulated a higher amount of HpD and did not differ significantly from BMG-1 cells with respect to intracellular localization. Photodynamically-induced cytogenetic damage and apoptosis were considerably higher in BMG-1 cells as compared to 4451 cells. The present results strongly suggest that manifestation of the photodynamically-induced lesions in the form of cytogenetic damage and apoptosis are among the important determinants of cellular sensitivity to HpD-PDT besides the photodynamic dose (intracellular concentration of the photosensitizer and the light dose). PMID- 12633984 TI - Fluorescence contrast and threshold limit: implications for photodynamic diagnosis of basal cell carcinoma. AB - This study was designed to evaluate what application time of delta-5 aminolaevulinic acid (ALA) results in highest contrast between tumour and normal skin, in the interval 1-4 h, when using photodynamic diagnosis (PDD) of basal cell carcinomas (BCC) located on the face. Moreover, a value of the demarcation limit has been derived based on the fluorescence variation in normal skin adjacent to the tumour. Forty patients were included in the study, randomly allocated to four different groups with varying ALA application time in the range 1-4 h. The contrast, defined as the ratio between the fluorescence intensity in ALA-treated tumour tissue and normal skin, was calculated for each patient, and the mean values in each group were evaluated as a function of ALA application time. In addition, the fluorescence intensity variation in ALA-treated normal skin adjacent to the tumour was assessed. The results from this study show a peak of the mean contrast values after 3 h ALA application, but due to large interpatient variation, the mean contrast did not differ significantly in the interval 2-4 h. After 2 h ALA application, the fluorescence intensity variation in the normal ALA-treated skin was found to be at a maximum, which suggests that 2 h ALA application is not preferable when using PDD. Based on data of the fluorescence variation in ALA-treated normal skin after 3 and 4 h ALA application, a tolerance interval was calculated implying that values above 1.4 times the mean normal fluorescence indicate an abnormal condition. This tolerance limit agrees well with results obtained in a former study. PMID- 12633985 TI - Treatment of cutaneous T cell lymphoma with extracorporeal photopheresis induces Fas-ligand expression on treated T cells, but does not suppress the expression of co-stimulatory molecules on monocytes. AB - Following extracorporeal photopheresis (ECP), lymphocytes become apoptotic and upregulate class I MHC antigenic peptides. Conversely, ECP treated monocytes demonstrate activation markers and have an increased avidity for the phagocytosis of apoptotic T cells. Processing of apoptotic T cells by monocytes, following ECP, is thought to induce an immunomodulatory response, which targets untreated, but clonal T cells. Recently we detected apoptotic lymphocytes immediately post ECP. Although enhanced CD95 (Fas) expression has been observed 24 h post ECP, CD95 and Fas-ligand (Fas-L) expression have not been determined at this very early apoptotic stage. Exposure of monocytes to UV has previously suppressed expression of the co-stimulatory molecules required for the presentation of processed antigens to T cells. Our data demonstrate no increase in CD95 or Fas-L expression on T cells tested immediately following ECP. However, the number of T cells expressing Fas-L significantly increased 24 h post ECP (P<0.005). The expression of the co-stimulatory molecules, CD54, CD80 and CD86, remained unaltered on monocytes treated by ECP. Although the mechanism responsible for early induction of lymphocyte apoptosis remains unclear, the later apoptosis involves Fas-L expression. The maintenance of co-stimulatory molecules, on treated monocytes, indicates that they retain the ability to induce an immunomodulatory response. PMID- 12633986 TI - Application of the biotin-dUTP chromosome labelling technique to study the role of 5-bromo-2'-deoxyuridine in the formation of UV-induced sister chromatid exchanges in CHO cells. AB - The role of 5-bromo-2'-deoxyuridine (BrdU) in the formation of sister chromatid exchanges (SCEs) in cells exposed to UV radiation was studied. Cells were unifilarily labelled (labelling of one strand of chromosomal DNA) with BrdU or biotin-16-2'-deoxyuridine (biotin-dU) and irradiated in G(1) phase of the cell cycle either with 254 nm, which is absorbed by all nucleobases including bromouracil (BrU) or with 313 nm radiation, which is predominantly absorbed by the BrU moiety. Elevated SCE frequencies were observed in cells irradiated at 254 nm (1.2 and 3.0 J m(-2)) which were pre-labelled with BrdU or biotin-dU. Following irradiation at 313 nm (38 and 96 J m(-2)) a statistically elevated SCE frequency was observed in cells pre-labelled with BrdU but not with biotin-dU. In cells pre-labelled with BrdU, UV-radiation at 254 nm was 50-80 times more effective in inducing SCEs than that at 313 nm. This result can be accounted for by the fact that in BrdU-DNA the cross-section for uracilyl radical and bromine atom formation is approximately 100-fold higher at 254 nm than that at 313 nm. Upon irradiation at 254 nm, BrdU had a strong sensitising effect on SCE induction: the SCE frequencies observed in cells pre-labelled with BrdU are approximately 6 times higher than in cells pre-labelled with biotin-dU. From this it follows that BrdU-induced damage is responsible for more than 80% of the SCEs formed in UV irradiated cells unifilarily labelled with BrdU. Based on photochemical considerations and the fact that chemical agents which form DNA interstrand cross-links are among the most potent inducers of SCEs, we propose that an interstrand cross-link may be the major lesion leading to SCEs in BrdU labelled cells. PMID- 12633987 TI - The riddle of the Sec1/Munc-18 proteins - new twists added to their interactions with SNAREs. AB - Sec1/Munc-18 (SM) proteins are essential for intracellular membrane fusion reactions. Most of them bind to membrane-associated soluble N-ethylmaleimide sensitive fusion (NSF)-attachment protein receptors (SNAREs) of the syntaxin subfamily but it is unclear whether regulating syntaxins is their primary role. Recent studies now have shown that the mechanism of syntaxin binding is not conserved, even though the structures of both protein families are. Amazing as this might be for those considering the evolution of conserved folds, it leaves the question of how SM proteins operate in membrane fusion unanswered. PMID- 12633988 TI - Expanding the physiological role of the hexameric DnaB helicase. AB - The hexameric DnaB protein is the Escherichia coli primary replicative helicase. However, recent studies indicate that this enzyme is capable of driving branch migration of a Holliday junction, acting more like an ATP-driven pump than a helicase. This offers an attractive possibility that the protein is directly involved in DNA recombination in vivo. Thus, the physiological role of the DnaB protein in DNA metabolism, and its modes of action, might be more complex than previously thought. PMID- 12633989 TI - An eight-cysteine-containing CFEM domain unique to a group of fungal membrane proteins. AB - CFEM, an eight cysteine-containing domain, has been identified by analyzing over 25 fungal sequences selected from database sequence searches. Features of CFEM suggest that it is a novel domain with characteristics distinct from known cysteine-rich domains. Some CFEM-containing proteins (e.g. Pth11 from Magnaporthe grisea) are proposed to have important roles in fungal pathogenesis. PMID- 12633990 TI - The NIT domain: a predicted nitrate-responsive module in bacterial sensory receptors. AB - A nitrate- and nitrite-sensing (NIT) domain as found in the NasR protein, has been detected in various receptor components of signal transduction pathways in different bacterial lineages. Cellular functions controlled by receptors that contain this novel domain include regulation of gene expression (transcription anti-terminators and histidine kinases), cell motility (chemotaxis receptors) and enzyme activity (diguanylate cyclases and phosphodiesterases). PMID- 12633991 TI - Historical review: an energy reservoir for mitosis, and its productive wake. AB - The realization that DNA is replicated during only a discrete part of interphase in most cells gave birth to the cell-cycle concept exactly half a century ago. But soon thereafter, a controversy arose as to whether a special reservoir of metabolic energy, built up at the end of interphase, was essential for the entry of cells into mitosis. After a period of intense debate, the 'energy reservoir' concept came to a sudden demise. But the notion that mitotic progression involves the cyclic build-up and consumption of key factors endured and, as we all now know so well, eventually triumphed. PMID- 12633992 TI - Translation initiation and viral tricks. AB - A variety of viral strategies are utilized for dominance of the host-cell protein synthetic machinery, optimization of viral mRNA translation and evasion of host cell antiviral responses that act at the translational level. Many viruses exploit regulated steps in the initiation of cellular protein synthesis to their own advantage. They have developed some rather unconventional means for mRNA translation, which were probably adapted from specialized cellular mRNA translation systems. Regardless of the type of translational tricks exploited, viruses typically ensure efficient viral translation, often at the expense of host-cell protein synthesis. PMID- 12633993 TI - Membrane proteins: the 'Wild West' of structural biology. AB - Historically, the task of determining the structure of membrane proteins has been hindered by experimental difficulties associated with their lipid-embedded domains. Here, we provide an overview of recently developed experimental and predictive tools that are changing our view of this largely unexplored territory the 'Wild West' of structural biology. Crystallography, single-particle methods and atomic force microscopy are being used to study huge membrane proteins with increasing detail. Solid-state nuclear magnetic resonance strategies provide orientational constraints for structure determination of transmembrane (TM) alpha helices and accurate measurements of intramolecular distances, even in very complex systems. Longer distance constraints are determined by site-directed spin labelling electron paramagnetic resonance, but current labelling strategies still constitute some limitation. Other methods, such as site-specific infrared dichroism, enable orientational analysis of TM alpha-helices in aligned bilayers and, combined with novel computational and predictive tools that use evolutionary conservation data, are being used to analyze TM alpha-helical bundles. PMID- 12633994 TI - Heparan sulfate proteoglycan as a plasma membrane carrier. AB - The plasma membrane defines the border of living cells and provides a barrier to extracellular components. Advances in molecular biology have resulted in the development of novel therapeutic strategies (e.g. gene therapy and cellular protein delivery) which rely on the entry of charged macromolecules into the intracellular compartment. Recent reports demonstrate an intriguing role for heparan sulfate proteoglycans in cellular internalization of viruses, basic peptides and polycation-nucleic-acid complexes and the possibility that they have important implications for gene transfer and protein delivery to mammalian cells. This review focuses on heparan sulfate proteoglycan as a plasma membrane carrier. PMID- 12633995 TI - New insights into fragile X syndrome: from molecules to neurobehaviors. AB - Fragile X syndrome - a common form of inherited mental retardation - is caused by the loss of the fragile X mental retardation 1 protein (FMRP). FMRP is an RNA binding protein which forms a messenger ribonucleoprotein (mRNP) complex that associates with translating polyribosomes. It has been proposed that FMRP is involved in synaptic plasticity through the regulation of mRNA transportation and translation. Recent advances in the identification of the mRNA ligands that are bound by FMRP, the RNA sequence and structure required for FMRP-RNA interaction, and the physiological consequences of FMRP deficiency in the brain are important steps towards understanding the molecular pathogenesis of fragile X syndrome, and learning and memory in general. PMID- 12633996 TI - mRNA display: ligand discovery, interaction analysis and beyond. AB - In vitro peptide and protein selection using mRNA display enables the discovery and directed evolution of new molecules from combinatorial libraries. These selected molecules can serve as tools to control and understand biological processes, enhance our understanding of molecular interactions and potentially treat disease in therapeutic applications. In mRNA display, mRNA molecules are covalently attached to the peptide or protein they encode. These mRNA-protein fusions enable in vitro selection of peptide and protein libraries of >10(13) different sequences. mRNA display has been used to discover novel peptide and protein ligands for RNA, small molecules and proteins, as well as to define cellular interaction partners of proteins and drugs. In addition, several unique applications are possible with mRNA display, including self-assembling protein chips and library construction with unnatural amino acids and chemically modified peptides. PMID- 12633997 TI - Ecological effects of various toxic agents on the aquatic microcosm in comparison with acute ionizing radiation. AB - The purpose of this study was an evaluation of the effect levels of various toxic agents compared with acute doses of ionizing radiation for the experimental model ecosystem, i.e., microcosm mimicking aquatic microbial communities. For this purpose, the authors used the microcosm consisting of populations of the flagellate alga Euglena gracilis as a producer, the ciliate protozoan Tetrahymena thermophila as a consumer and the bacterium Escherichia coli as a decomposer. Effects of aluminum and copper on the microcosm were investigated in this study, while effects of gamma-rays, ultraviolet radiation, acidification, manganese, nickel and gadolinium were reported in previous studies. The microcosm could detect not only the direct effects of these agents but also the community-level effects due to the interspecies interactions or the interactions between organisms and toxic agents. The authors evaluated doses or concentrations of each toxic agent which had the following effects on the microcosm: (1) no effects; (2) recognizable effects, i.e., decrease or increase in the cell densities of at least one species; (3) severe effects, i.e., extinction of one or two species; and (4) destructive effects, i.e., extinction of all species. The resulting effects data will contribute to an ecological risk assessment of the toxic agents compared with acute doses of ionizing radiation. PMID- 12633998 TI - Monitoring and modelling of indoor radon concentrations in a multi-storey building at Mumbai, India. AB - Radon (Rn(222)) levels in an indoor atmosphere of a multi-storey building at Mumbai have been measured for one year covering all the four seasons. Monitoring was carried out using the time-integrated passive detector technique, using Kodak 115 type Solid State Nuclear Track Detector (SSNTD) films of 2.5x2.5 cm size. Measured indoor radon levels showed a decreasing trend with height with concentration ranging from 41 Bq m(-3) at ground floor level to 15 Bq m(-3) at 19th floor level. Using the dose conversion factors, the inhalation dose due to breathing of radon gas is estimated to be 1.03 mSv y(-1) at the ground floor to 0.38 mSv y(-1) at the 19th floor level. Measured indoor radon concentrations on each floor were compared with the computed values using a mathematical model. The agreement between measured values and calculated values of indoor concentrations at different floors was very good within the limitations of various field parameter values. PMID- 12633999 TI - Detection of 210Po on filter papers 16 years after use for the collection of short-lived radon progeny in a room. AB - Radon gas was allowed to accumulate in its radium source and then injected into a 36 m(3) test room, resulting in an initial radon concentration of 15 kBq m(-3). Filter papers were used to collect the short-lived radon progeny and thus to measure the Potential Alpha Energy Concentration (PAEC) in-situ in the year 1984 at different times and conditions according to the experimental design. The radon progeny collected on the filter papers were studied as a function of aerosol particle concentration ranging from 10(2)-10(5) particles cm(-3) in three different experiments. The highest aerosol particle concentration was generated by indoor cigarette smoking. Those filters were stored after the experiment, and were used after 16 years to study the activity of the radon long-lived alpha emitter progeny, (210)Po (T(1/2)=138 days). This isotope is separated from the short-lived progeny by (210)Pb beta emitter with 22.3 years half-life. After 16 years' storage of these filters, each filter paper was sandwiched and wrapped between two CR-39 nuclear track detectors, to put the detectors in contact with the surfaces of different filters, for 337 days. Correlation between the PAEC measured using filter papers in the year 1984 and the activity of long-lived alpha emitter (210)Po on the same filter papers measured in the year 2000 were studied. The results of the (210)Po activity showed a very good correlation of 0.92 with the PAEC 16 years ago. The results also depict that the PAEC and (210)Po activity in indoor air increased with the increase of aerosol particle concentration, which shows the attachment of short-lived radon progeny with the aerosol particles. The experiment proves that indoor cigarette smoking is a major source of aerosol particles carrying radon progeny and, thus, indoor cigarette smoking is an additional source of internal radiation hazard to the occupants whether smoker or non-smoker. PMID- 12634000 TI - An inventory of long-lived radionuclides residual from underground nuclear testing at the Nevada test site, 1951-1992. AB - An inventory of long-lived radionuclides produced by 828 underground nuclear tests conducted at the Nevada test site (NTS) from 1951 to 1992 includes residual tritium, fission products, actinides, and activation products. Recently, the US Department of Energy approved the declassification of the NTS radionuclide inventory by principal geographic test centers. This permits unclassified publication of radionuclide totals for the Yucca Flat, Pahute Mesa-Area 19, Pahute Mesa-Area 20, Frenchman Flat, and Rainier Mesa/Shoshone Mountain testing locations. Activities are reported as of September 23, 1992, the date of the last underground nuclear test conducted at the NTS, and September 23, 2492, after 500 years of radioactive decay. The availability of these data affords an opportunity for the analysis of the radiologic source term within the boundaries of local hydrogeologic units and provides insight to where radionuclides are sited relative to potential exposure pathways. PMID- 12634001 TI - Radioactivity concentrations of 40K, 134Cs, 137Cs, 90Sr, 241Am, 238Pu and 239+240Pu radionuclides in Jordanian soil samples. AB - In November 2000, surface and core soil samples were collected from different regions of Jordan. The samples were analyzed by direct gamma spectrometry and combined radiochemical separation procedure to quantify (40)K, (134)Cs, (137)Cs, (90)Sr, (241)Am, (238)Pu and (239+240)Pu radioactivity. Concentrations (Bq.kg(-1) dry weight) have been observed to vary in the range 1.5-2.6 for (134)Cs, 2.8-11.4 for (90)Sr, and 0.13-0.48 for (241)Am, 0.016-0.062 for (238)Pu, 0.28-1.01 for (239+240)Pu and 155-543 for (40)K. The typical concentration of (137)Cs found in topsoils (0-2 cm) ranged in 7.5-576 Bq.kg(-1), dry weight. These values were greater than those observed in samples taken at greater depths (up to 32 cm). Activity ratios of (134)Cs/(137)Cs, (90)Sr/(137)Cs, (239+240)Pu/(137)Cs, (238)Pu/(137)Cs, (241)Am/(137)Cs, (239+240)Pu/(238)Pu and (241)Am /(238)Pu have mean values of 0.0049 (R=1), 0.29 (R=0.76), 0.41 (R=0.90), 0.39 (R=0.85), 0.41 (R=0.88), 7.72 (R=0.97) and 16.66 (R=0.98), respectively. The underlying concentrations were correlated and relatively higher than those reported in neighboring countries. One moss sample, as a biomonitor indicator, was measured and evaluated along with the soil samples. Its data showed higher concentrations of all measured radionuclides due to accumulations over years. The depth distribution of the fission product (137)Cs and the total deposition (Bq.m(-2)) were also studied in selected samples. Estimations of the annual effective dose equivalent due to (137)Cs-soil contamination showed values up to more than 200 microSv. PMID- 12634002 TI - 238U series isotopes and 232Th in carbonates and black shales from the Lesser Himalaya: implications to dissolved uranium abundances in Ganga-Indus source waters. AB - 238U and (232)Th concentrations and the extent of (238)U-(234)U-(230)Th radioactive equilibrium have been measured in a suite of Precambrian carbonates and black shales from the Lesser Himalaya. These measurements were made to determine their abundances in these deposits, their contributions to dissolved uranium budget of the headwaters of the Ganga and the Indus in the Himalaya and to assess the impact of weathering on (238)U-(234)U-(230)Th radioactive equilibrium in them. (238)U concentrations in Precambrian carbonates range from 0.06 to 2.07 microg g(-1). The 'mean' U/Ca in these carbonates is 2.9 ng U mg(-1) Ca. This ratio, coupled with the assumption that all Ca in the Ganga-Indus headwaters is of carbonate origin and that U and Ca behave conservatively in rivers after their release from carbonates, provides an upper limit on the U contribution from these carbonates, to be a few percent of dissolved uranium in rivers. There are, however, a few streams with low uranium concentrations, for which the carbonate contribution could be much higher. These results suggest that Precambrian carbonates make only minor contributions to the uranium budget of the Ganga-Indus headwaters in the Himalaya on a basin wide scale, however, they could be important for particular streams. Similar estimates of silicate contribution to uranium budget of these rivers using U/Na in silicates and Na* (Na corrected for cyclic and halite contributions) in river waters show that silicates can contribute significantly (approximately 40% on average) to their U balance. If, however, much of the uranium in these silicates is associated with weathering resistant minerals, then the estimated silicate uranium component would be upper limits. Uranium concentration in black shales averages about 37 microg g(-1). Based on this concentration, supply of U from at least approximately 50 mg of black shales per liter of river water is needed to balance the average river water U concentration, 1.7 microg L(-1) in the Ganga-Indus headwaters. Data on the abundance and distribution of black shales in their drainage basin are needed to test if this requirement can be met. (234)U/(238)U activity ratios in both carbonates and black shales are at or near equilibrium, thus preferential mobilization of (234)U from these deposits, if any, is within analytical uncertainties. (230)Th is equivalent to or in excess of (238)U in most of the carbonates. (230)Th/(238)U>1 indicates that during weathering, uranium is lost preferentially over Th. (232)Th concentrations in carbonates are generally quite low, <0.5 microg g(-1), though with a wide range, 0.01-4.8 microg g(-1). The variation in its concentrations seem to be regulated by aluminosilicate content of the carbonates as evident from the strong positive correlation between (232)Th and Al. PMID- 12634003 TI - Don't shoot the messenger. PMID- 12634004 TI - Gleevec: tailoring to fit. PMID- 12634005 TI - Smart bombs: the next generation of PDT. PMID- 12634008 TI - Pat Price discusses the potential of molecular imaging for drug development. Interview by Joanna Owens. PMID- 12634009 TI - Drug discovery perspective of chemical-driven mouse mutagenesis. PMID- 12634010 TI - ENU mutagenesis for pharma. PMID- 12634011 TI - Advancing technologies for accelerated drug development. AB - The 1st International Drug Discovery and Development Summit--From Lead to Drug in Five Years (December 2-5, 2002, Honolulu, HI, USA) focused on technologies with potential to accelerate the drug development process. PMID- 12634012 TI - Delivering the power of discovery in large pharmaceutical organizations. AB - Increasing the output from discovery is currently a major objective for the pharmaceutical industry aimed at reversing recent downward trends in productivity. Although significant attention has been focused on innovative assay and process technologies, these only address specific points in the discovery process. Little effort has been made to manage the multiple interconnected steps within discovery effectively. This manifests itself in low utilization rates of component elements and low expectations of delivery to any agreed timescale. PMID- 12634013 TI - Rational design and engineering of therapeutic proteins. AB - An increasing number of engineered protein therapeutics are currently being developed, tested in clinical trials and marketed for use. Many of these proteins arose out of hit-and-miss efforts to discover specific mutations, fusion partners or chemical modifications that confer desired properties. Through these efforts, several useful strategies have emerged for rational optimization of therapeutic candidates. The controlled manipulation of the physical, chemical and biological properties of proteins enabled by structure-based simulation is now being used to refine established rational engineering approaches and to advance new strategies. These methods provide clear, hypothesis-driven routes to solve problems that plague many proteins and to create novel mechanisms of action. We anticipate that rational protein engineering will shape the field of protein therapeutics dramatically by improving existing products and enabling the development of novel therapeutic agents. PMID- 12634014 TI - The impact of nucleic acid secondary structure on PNA hybridization. AB - Hybridization of oligonucleotides and their analogues to complementary DNA or RNA sequences is complicated by the presence of secondary and tertiary structure in the target. In particular, folding of the target nucleic acid imposes substantial thermodynamic penalties to hybridization. Slower kinetics for hybridization can also be observed, relative to an unstructured target. The development of high affinity oligonucleotide analogues such as peptide nucleic acid (PNA) can compensate for the thermodynamic and kinetic barriers to hybridization. Examples of structured targets successfully hybridized by PNA oligomers include DNA duplexes, DNA hairpins, DNA quadruplexes and an RNA hairpin embedded within a mRNA. PMID- 12634016 TI - Methods to assess and improve the physical parameters associated with fall risk in older adults. AB - BACKGROUND: Falls are common among older adults. Many physical parameters including reduced postural stability, decreased dynamic balance, gait disorders, strength deficits, difficulty standing from a chair, and other impairments have been shown to be strongly associated with fall risk in the elderly. ASSESSMENTS: To identify those at risk for falls, tools that accurately measure physical performance parameters associated with falls are essential. Several tools are available to measure these parameters including clinical evaluations, functional performance tests, and questionnaires. The article describes many of the tools that can be used to evaluate the physical parameters associated with fall risk in older adults. CONCLUSIONS: The described instruments can help in identifying those who are most likely to fall, and those who would benefit from targeted interventions. The final part of the article includes a brief discussion of the potential role of exercise training interventions to improve these physical parameters and prevent falls. PMID- 12634017 TI - Effectiveness of a nationally implemented smoking cessation guideline on provider and patient practices. AB - BACKGROUND: The Agency for Health Care Policy and Research (AHCPR) smoking cessation guideline outlines a set of recommendations for physicians to follow in daily practice. However, the effectiveness of this guideline has not been reported. The goal of this project was to evaluate the effect of the AHCPR smoking cessation guideline on provider practices with smokers and on patient smoking rates. METHODS: Patient survey and chart review data from 138 Veterans Administration (VA) acute care medical centers with outpatient facilities were examined. Data were available from both sources in 1996, 1997, and 1998. At the midpoint of this period (1997), the VA recommended the AHCPR smoking cessation clinical practice guideline for implementation throughout the VA healthcare system. RESULTS: From 1996 to 1998, both the chart audit and the patient survey showed a significant increase in the percentage of patients in the VA who were counseled about smoking and a significant decrease in the percentage of patients who smoke. CONCLUSIONS: Because the VA tied the guideline implementation to report cards and other performance-enhancing measures, guideline adherence may have been maximized in this setting. These findings suggest that healthcare systems should take an integrated approach to guideline implementation. PMID- 12634018 TI - Smoking visibility, perceived acceptability, and frequency in various locations among youth and adults. AB - OBJECTIVES: This study tests whether there is an association between high visibility of smoking, perceived acceptability of smoking, and where youth smoke. METHODS: Surveys of 9,762 students in grades 8-10 and 1,586 parents in 15 Minnesota communities asked about the frequency of and opinions of adult and youth smoking in various indoor and outdoor public places. Chi-square analysis and ANOVAs compared smokers and nonsmokers. RESULTS: More smoking than nonsmoking youth reported often seeing adults and teens smoking in all locations. Forty-two percent of students often noticed adults and 35% often noticed teens smoking outdoors, also the most frequent location where teens report smoking. Students perceived adult and teen smoking as more acceptable in restaurants, recreation centers, and outdoor gathering places. More student smokers than nonsmokers believe that smoking is acceptable for both adults and teens. Of the parent sample, 52% often observed adults and 36% often observed youth smoking outdoors. Nonsmoking parents observed adult and teen smoking more often than parents who smoke. CONCLUSIONS: The data support an association between the frequency that youth observe smoking in various locations and the perception that smoking is socially acceptable by smoking status. Policies that restrict smoking in various locations will reduce both visibility and perceived acceptability of smoking in those locations. PMID- 12634019 TI - Determinants of salivary cotinine concentrations in Chinese male smokers. AB - BACKGROUND: Identifying factors that affect cotinine levels in smokers may be useful for smoking cessation programs. Our aims were to characterize the distribution of salivary cotinine levels in Chinese smokers and to investigate factors that influence cotinine concentrations. METHODS: In a cross-sectional study, 600 Chinese adult smokers answered a questionnaire on smoking habits and provided a saliva sample for cotinine analysis. Modification of the relation between number of cigarettes smoked and cotinine concentration by individual characteristics, smoking behavior, and type of tobacco was evaluated. RESULTS: Quadratic model provided the best fit for the relation between number of cigarettes smoked in the previous 24 hours and salivary cotinine concentration. Among those smoking up to 20 cigarettes, the median cotinine concentration was higher among younger subjects, those smoking cigarettes without filter and regular rather than light cigarettes, and those inhaling frequently and deeply. Such trends were not observed among heavier smokers. The increase in cotinine per cigarette tended to be larger in those with lower median cotinine level. CONCLUSIONS: Our findings show that smoking behavior-related factors modify the relation between number of cigarettes smoked and salivary cotinine concentration. This suggests that smokers may regulate their smoking behavior to achieve a certain optimum nicotine level. PMID- 12634021 TI - The public health burden of material deprivation: excess mortality in leading causes of death in Spain. AB - OBJECTIVE: The aim of the study is to investigate the burden of deprivation associated excess of deaths by 10 leading causes of death in each gender and two age groups. DESIGN: A small-area ecological study using two indices of material deprivation drawn from 1991 census and mortality data aggregated for 1987-1995 was undertaken. SETTING: 2218 small areas in Spain. Data are presented by region. MAIN OUTCOME MEASURES: The 10 leading causes of death in each gender and two age groups were determined. RESULTS: The deprivation-associated excess mortality in the top 10 leading causes of death was 14.4 and 11% for males and females, respectively. Excess mortality in the 9-year period studied accounted for more than 222,000 deaths. A large proportion of excess deaths was found in causes related to smoking and alcohol consumption among males and diet-related causes of death among females. The most deprived regions within the country showed the highest mortality risk. CONCLUSIONS: Material deprivation appears to be manifested in high mortality rates differently by gender and region. It imposes a significant public health burden that demands urgently attention and action by health policy-makers. PMID- 12634020 TI - The impact of financial incentives and a patient registry on preventive care quality: increasing provider adherence to evidence-based smoking cessation practice guidelines. AB - BACKGROUND: This study tested the effects of two organizational support processes, the provision of financial incentives for superior clinical performance and the availability of a patient (smoker) registry and proactive telephone support system for smoking cessation, on provider adherence to accepted practice guidelines and associated patient outcomes. METHODS: Forty clinics of a large multispecialty medical group practice providing primary care services were randomly allocated to study conditions. Fifteen clinics each were assigned to the experimental conditions "control" (distribution of printed versions of smoking cessation guidelines) and "incentive" (financial incentive pay-out for reaching preset clinical performance targets). Ten clinics were randomized to receive financial incentives combined with access to a centralized patient registry and intervention system ("registry"). Main outcome measures were adherence to smoking cessation clinical practice guidelines and patients' smoking cessation behaviors. RESULTS: Patients' tobacco use status was statistically significant (P < 0.01) more frequently identified in clinics with the opportunity for incentives and access to a registry than in clinics in the control condition. Patients visiting registry clinics accessed counseling programs statistically significantly more often (P < 0.001) than patients receiving care in the control condition. Other endpoints did not statistically significantly differ between the experimental conditions. CONCLUSIONS: The impact of financial incentives and a patient registry/intervention system in improving smoking cessation clinical practices and patient behaviors was mixed. Additional research is needed to identify conditions under which such organizational support processes result in significant health care quality improvement and warrant the investment. PMID- 12634022 TI - A lifestyle intervention improves plasma insulin levels among Native American high school youth. AB - BACKGROUND: Worldwide, type 2 diabetes prevalence is increasing, with Native American populations particularly at risk. The Zuni Pueblo, with a history of wellness activities, volunteered to test the feasibility and efficacy of a high school-based diabetes prevention intervention. METHODS: This school-based intervention used a multiple cross-sectional design to evaluate outcome measures at 0, 1.5, and 3 years against an Anglo comparison group. The Zuni high school diabetes prevention program included an educational component targeting decreased consumption of sugared beverages, knowledge of diabetes risk factors, and a youth oriented fitness center. Main outcome measures were plasma glucose and insulin measured fasting and 30 min after a 75-g glucose challenge. RESULTS: Plasma glucose levels were normal at baseline for Zuni (n = 72) and Anglo (n = 37) youth and did not significantly change throughout the study. At baseline, fasting and 30-min plasma insulin levels were significantly elevated for Zuni youth; they showed significant steady declines for both males and females throughout the study (P = 0.06 to P = 0.000 for trends using quantile regression). By Year 3, values for Zuni males (n = 29) equaled Anglo comparison values, while Zuni female (n = 26) values had declined but were still higher than Anglo comparison values. CONCLUSIONS: Among at-risk youth, an environmentally based lifestyle intervention may significantly suppress markers of type 2 diabetes risk. PMID- 12634023 TI - Hip-Hop to Health Jr., an obesity prevention program for minority preschool children: baseline characteristics of participants. AB - BACKGROUND: The prevalence of obesity in the United States is a significant public health problem. Many obesity-related risk factors are more prevalent in minority populations. Given the recalcitrant nature of weight loss interventions for adults, prevention of overweight and obesity has become a high priority. The present study reports baseline data from an obesity prevention intervention developed for minority preschool children. METHODS: Hip-Hop to Health Jr. is a 5 year randomized controlled intervention that targets 3- to 5-year-old minority children enrolled in 24 Head Start programs. Our primary aim is to test the effect of the intervention on change in body mass index. Data were collected on sociodemographic, anthropometric, behavioral, and cognitive variables for the children and parents at baseline. RESULTS: Participants included 416 black children, 337 black parents, 362 Latino children, and 309 Latino parents. Using body mass index for age and sex > or = the 95th percentile as the definition of overweight, 15% of the black children and 28% of the Latino children were overweight. More than 75% of the parents were either overweight or obese. DISCUSSION: The development of interventions to effectively prevent or control obesity early in life is crucial. These data highlight the escalating problem of weight control in minority populations. PMID- 12634024 TI - Associations of weight loss and changes in fat distribution with the remission of hypertension in a bi-ethnic cohort: the Atherosclerosis Risk in Communities Study. AB - BACKGROUND: To examine associations of weight loss and changes in fat distribution with changes in blood pressure and the remission of hypertension in a community-based sample. METHODS: Participants were 3245 white and African American men and women, 45-64 years of age, who participated in the Atherosclerosis Risk in Communities Study over an average of 9 years. Mixed models analyses were used to examine the associations of weight loss and changes in fat distribution with changes in blood pressure. Proportional hazard models with time-dependent covariates were used to examine the associations of weight loss and changes in fat distribution with the remission of hypertension. RESULTS: Weight loss was associated with a decrease in systolic blood pressure and diastolic blood pressure and with an increased rate of remission of hypertension. Hazard ratios of the remission of hypertension associated with 1-kg increment in annual weight loss were 2.04 (95% confidence interval [CI]: 1.62-2.59), 1.38 (95% CI: 1.14-1.67), 1.84 (95% CI: 1.47-2.29), and 1.53 (95% CI: 1.14-2.05) for white women, African-American women, white men, and African-American men, respectively. Changes in fat distribution were associated with the remission of hypertension in younger (45-54 years) participants. CONCLUSIONS: Weight loss was associated with a decrease in blood pressure and with remission of hypertension in white and African-American men and women. PMID- 12634025 TI - A randomized controlled trial of a public health nurse directed treatment program for rural patients with high blood cholesterol. AB - BACKGROUND: Many rural residents do not have access to high-quality nutrition counseling for high blood cholesterol. The objective of this study was to assess the effectiveness of an intervention program designed to facilitate dietary counseling for hypercholesterolemia by rural public health nurses. METHODS: Eight health departments (216 participants) were randomized to give the special intervention (SI) and nine (252 participants) to give the minimal intervention (MI). The SI consisted of three individual diet counseling sessions given by a public health nurse, using a structured dietary intervention (Food for Heart Program), referral to a nutritionist if lipid goals were not achieved at 3-month follow-up, and a reinforcement phone call and newsletters. Diet was assessed by the Dietary Risk Assessment (DRA), a validated food frequency questionnaire, at baseline, 3-, and 12-month follow-up; blood lipids and weight were assessed at baseline, 3-, 6-, and 12-month follow-up. RESULTS: Participants were largely female (71%), older (mean age 55), and white (80%). At 3-month follow-up, the average reduction (indicating dietary improvement) in total Dietary Risk Assessment score was 3.7 units greater in the SI group (95% confidence interval [CI] 1.9 to 5.5, P = 0.0006), while both groups experienced a similar reduction in blood cholesterol, 14.1 mg/dL (0.37 mmol/L) for SI and 14.5 mg/dL (0.38 mmol/L) for minimal intervention group (difference -0.4 mg/dL [-0.010 mmol/L], 95% CI -12.5 to 11.7 [-0.32 to 0.30], P = 0.9). At 12-month follow-up, the reduction in total Dietary Risk Assessment score was 2.1 units greater in the SI group (95% CI 0.8 to 3.5, P = 0.005), while the reduction in blood cholesterol was similar in both groups, 18.4 mg/dL (0.48 mmol/L) for SI and 15.6 mg/dL (0.40 mmol/L) for minimal intervention group (difference 2.8 mg/dL [0.07 mmol/L], 95% CI -7.5 to 13.1 [-0.19 to 0.34], P = 0.6). During follow-up, weight loss was greater in the SI group; the difference between groups was statistically significant at 3 (1.9 lb [0.86 kg], 95% CI 0.3 to 3.4 [0.14 to 1.55], P = 0.022) and 6 months (2.1 lb [0.95 kg], 95% CI 0.1 to 4.1 [0.04 to 1.86], P = 0.04). At 12 months, the difference was not significant (1.6 lb [0.73 kg], 95% CI -0.05 to 3.7 [-0.02 to 1.68], P = 0.13). CONCLUSIONS: Improvement in self-reported dietary intake was significantly greater in the SI group, while reduction in blood cholesterol was similar in both groups. PMID- 12634026 TI - A national survey of primary care physicians' colorectal cancer screening recommendations and practices. AB - BACKGROUND: National data on providers' colorectal cancer (CRC) screening knowledge, attitudes, and practices are sparse. This study assessed primary care physicians' (PCPs') beliefs about the effectiveness of CRC screening, their recommendations for screening, their perceptions of the influence of published guidelines on their CRC screening recommendations, and how they conduct CRC screening in their clinical practices. METHODS: A questionnaire was administered to a nationally representative sample of practicing PCPs. Of 1718 eligible physicians, 1235 (72%) responded. RESULTS: Only 2% of PCPs said they did not recommend CRC screening. Over 80% indicated that they most often recommend CRC screening with fecal occult blood testing and/or flexible sigmoidoscopy, although colonoscopy was perceived as the more effective screening modality. Nearly two thirds of obstetrician/gynecologists and one-fourth of other practitioners reported conducting fecal occult blood testing exclusively by digital rectal exam. Only 29% of PCPs said they perform sigmoidoscopy. Estimated volumes of ordering, performing, or referring for CRC screening were low, and <20% reported that three-fourths or more of their older patients were up to date with CRC screening as recommended by the physician. Many PCPs reported recommending CRC screening at nonstandard starting ages or too-frequent intervals. CONCLUSIONS: Awareness of CRC screening among PCPs in the United States is high. However, knowledge gaps about the timing and frequency of screening and suboptimal screening delivery were evident. PMID- 12634027 TI - Cognitive motivations associated with screening mammography in Cyprus. AB - BACKGROUND: This study examined the relationship between cognitive motivations, demographic characteristics, related preventive health behaviors, and intention to obtain a first screening mammogram among Greek Cypriot women. METHODS: This cross-sectional study took place at the outpatient clinics of the General Hospital of Nicosia during the fall of 1999. The participants were 293 women asymptomatic of breast cancer and with no previous mammographic experience. The Theory of Planned Behavior in conjunction with the construct of self-efficacy was used in the development of the questionnaire. The assessment also included demographic information, practice of other preventive health behaviors, and knowledge of breast cancer screening. RESULTS: Self-efficacy was the most significant determinant of initial screening mammography, followed by normative beliefs associated with the physician, family and close friends, and perceived behavioral control beliefs related to barriers to obtaining a mammogram. Educational level, time of last clinical breast examination, and status of breast self-examination were also strongly implicated in the explanation of initial screening mammography. CONCLUSIONS: Insights into specific salient motivational cognitions and certain demographic characteristics or related preventive health behaviors are crucial to the development and implementation of effective intervention strategies. PMID- 12634028 TI - The relationship of symptoms and psychological factors to delay in seeking medical care for breast symptoms. AB - BACKGROUND: The psychological processes involved in the delay between noticing breast symptoms and seeking medical care are not well understood. METHODS: We evaluated 85 women referred to a specialist breast clinic prior to their clinic appointment. We assessed the relationship between delay and the type of breast symptom, immediate emotional response to the symptom, perceived risk of breast cancer, fear of breast cancer treatment, and disclosure of the breast symptom to others. RESULTS: Delay was unrelated to demographic factors but was related to the type of breast symptom; women who had a breast lump waited a significantly shorter time period before visiting the doctor than those without a breast lump. Initial symptom distress on the discovery of the breast symptom was also significantly related to delay. Knowledge of a friend or family member with breast cancer, perceived risk of breast cancer and fear of breast cancer treatment, and disclosure of the symptom to a partner or other person were all unrelated to delay. CONCLUSIONS: The results show the importance of the type of symptom and initial emotional distress in delay and highlight the importance of widening public perceptions of breast symptoms other than breast lumps in order to reduce delay times. PMID- 12634029 TI - Antioxidant supplement use in Women's Health Initiative participants. AB - BACKGROUND: Although antioxidant supplements are widely available and commonly used in the United States, there is a lack of detailed information on their use, including types of supplements used, doses, duration of use, and changes in use over time. METHODS: Antioxidant supplement use was assessed in participants in the Clinical Trial (n = 68,133) and Observational Study (n = 93,676) of the Women's Health Initiative. In-person interviews and a computerized inventory procedure were used to collect data on supplement use during baseline clinic visits from 1993 through 1998. RESULTS: Antioxidant supplements were widely consumed. For example, 55.5% of participants reported taking supplemental vitamin C in some form. Supplement use was positively associated with age, education, and physical activity. Most antioxidants were consumed through multivitamins. However, high doses were commonly consumed from single supplements, with 43.9% using single vitamin C supplements taking >500 mg and 86.1% using single vitamin E supplements taking >200 IU daily. Except for beta-carotene, there were increases in the use of all supplements from 1993 to 1998. CONCLUSIONS: This report demonstrates the feasibility of collecting comprehensive dietary supplement use data in large studies. These data may aid in the design of supplement use questionnaires, which could help to prevent misclassification error in epidemiologic studies of diet and disease. PMID- 12634030 TI - Where has evidence come from, what do we have now and what will we have in the future? PMID- 12634031 TI - The future of midwifery education. AB - In this paper I will examine the history shared by our profession and review social changes and reproductive health needs that have had an effect on our current practice. I will propose a model that meets current needs and will build the vision for our future education. In conclusion I will focus on the strategies, tasks and commitments needed to build a common future for midwifery education throughout the world. PMID- 12634032 TI - Midwives and women together for the family of the world: the place of research. AB - In this paper, I explore the place of research in the theme 'Midwives and women: together for the family of the world'. I begin by examining each of the concepts involved. What is the family of the world? Midwives: what do they contribute to the family of the world? Women: what do they contribute to the family of the world? Putting midwives and women together what is the impact on the family of the world. What is the place of research and what is its impact? I build an argument to show that midwifery research covers social, cultural and political spheres. It breaks the barriers between midwives and women and helps create a true partnership where there is interactivity rather than just one-way information giving from midwife to woman. Once midwives interact with women, they are more likely to persuade them and the world around the women to change their way of doing things. By evaluating what they do using research, midwives are able to capture the safe motherhood success stories and effect replication where needed. With research, midwives become unified, assertive, and mutually supportive in order to reinforce professional identity, accountability and autonomy in the delivery of quality care for women and their families. The family of the world therefore has a ready source of care and support, leadership, knowledge and policies that have been tried and tested to be effective in contributing to the health of women and their families. PMID- 12634033 TI - A case/comparison study in the Eastern Region of Ghana on the effects of incorporating selected reproductive health services on family planning services. AB - OBJECTIVE: to assess the impact on the provision of family planning (FP) services when FP providers were also trained to provide additional, selected, reproductive health services. DESIGN: case/comparison study. PARTICIPANTS AND SETTINGS: twenty four FP service delivery points in which training in sexually transmitted infection prevention and control services or post-abortion care services had been initiated (case facilities), were compared to 19 control facilities in which similar provider training had not yet been targeted. All settings were located in the Eastern Region of Ghana. MEASUREMENTS: service statistics for three study years (1996-1998) were reviewed. Structured interviews with providers, managers and clients provided qualitative data concerning impact and satisfaction. FINDINGS: case facilities which had integrated these additional reproductive health (RH) services experienced consistently higher numbers of clients and the total number of clients receiving FP services increased over time. There was also a statistically significant increase in continuing FP clients within case facilities. In contrast, the number of FP clients serviced in the comparison area remained basically unchanged over time. KEY CONCLUSIONS: interviews conducted with providers and managers in both types of settings indicated strong support for receipt of training to provide these integrated services and a request for additional training in an even broader array of RH and adult/child services. Clients also perceived the benefit of additional RH services and perceived these services to be of high quality. IMPLICATIONS FOR PRACTICE: expanding the repertoire of clinical skills of FP providers, enabling these practitioners to render RH services that augment basic FP services, has the potential to increase the number of new and continuing FP clients, and increases the satisfaction of both providers and consumers with respect to these services. PMID- 12634034 TI - Perceptions and experiences of motherhood, health and the husband's role among Thai women in Australia. AB - OBJECTIVE: to discuss the perceptions and experiences of motherhood among Thai immigrant women in Australia. DESIGN: ethnographic interviews and participant observation were used to elicit information. SETTING: Melbourne Metropolitan Area, Victoria, Australia. PARTICIPANTS: 30 Thai women who are now living in Australia. FINDINGS: becoming a mother meant different things to these women. Motherhood had both positive and negative aspects. Women believed that their health became worse as a result of becoming a mother. The contributing causes of their health status were multi-faceted, including their age, the demanding tasks of motherhood, the depletion of their strength and energy from childbirth, and in not following Thai traditional confinement practices. The findings in this paper support previous literature that motherhood has a profound effect on the life of many women. We, however, found that cultural beliefs and practices have an effect on women's perceptions and experience of motherhood. We also found that the 'presence' of the husband when a woman becomes a mother is important. The ethnicity of their husbands also played a significant role in their motherhood role and the ways in which they mothered their children. IMPLICATIONS FOR PRACTICE: understanding the social and cultural environment in which immigrant women try to be a mother is important if we are to provide culturally sensitive assistance and care to women who choose to become a mother in their new homeland. PMID- 12634035 TI - A qualitative study exploring junior paediatricians', midwives', GPs' and mothers' experiences and views of the examination of the newborn baby. AB - OBJECTIVE: to explore the experiences and attitudes of midwives, junior paediatricians (SHOs), GPs, and mothers to the examination of the newborn baby. To provide an appreciation of their views on several issues, in particular the purpose and value of the examination, who is thought to be appropriate to carry it out and when and where it should take place. DESIGN: qualitative using semi structured interviews, which were exploratory and interactive, in order to examine the range and diversity of experiences and attitudes to the neonatal examination. SETTING: South-east England. PARTICIPANTS: four samples were purposefully selected to include ten each of midwives, SHOs, GPs and recently delivered mothers. SHOs were currently working in paediatric departments of a district general hospital or teaching hospital and their experience of conducting examinations of the newborn baby ranged from several months to several years. Midwives included both those trained in the examination and currently conducting examinations, and those not so trained and not carrying out the examination. Most of the midwives had been qualified for over ten years and had a wide range of clinical experience in hospital and community settings. The GPs were from ten practices in two Health Authorities and all had some experience of conducting neonatal examinations. Of the mothers, a few had had their babies examined at home by midwives, others in hospital by an SHO. Mothers included those with a family history of problems relevant to the examination, those with previous pregnancy complications and others with no problems or complications. Some were first-time mothers. FINDINGS: all groups perceived the examination to be a useful screening tool providing reassurance to parents. They considered both midwives and SHOs to be appropriate professionals to carry out the examination, if adequately trained. Most thought that midwives have a better rapport with mothers, are able to provide continuity of care and more often discuss health care issues than do SHOs. Few SHOs reported receiving any formal training in the examination of the newborn baby. IMPLICATIONS FOR PRACTICE: the extension of the practice of midwives examining the newborn baby following relevant training would be acceptable to all stakeholders. The implications of increased demands on the midwives' workload may need to be considered. PMID- 12634036 TI - An analysis of Simpson's notebook data on the wet nurse. AB - OBJECTIVE: to understand the meaning of the qualitative data included in the Notebook of wet nurses kept by James Young Simpson. DESIGN: quantitative and qualitative analysis of data in an historical document. SETTING: a list of wet nurses kept by a 'Professor of Midwifery' in mid-19th century Edinburgh. PARTICIPANTS: the Notebook lists the names and other details of 749 women. FINDINGS: the Notebook indicates how the wet nurse was recruited, the implications for her baby, how she negotiated her role and the decision-making around her recruitment. KEY CONCLUSIONS: the ambiguity of this medical pioneer's decision-making is demonstrated. Simpson's scientific credentials may have featured much rhetoric. While in the forefront of many obstetric and medical developments, Simpson was regressive in his support for wet nursing. The social input into the selection of the wet nurse has not been identified previously. IMPLICATIONS FOR PRACTICE: the woman's ability to negotiate her terms of employment emerges. The social determinants of baby feeding decisions, identified in this document, have assumed greater significance since the time that this Notebook was written. PMID- 12634037 TI - A qualitative study exploring midwives' perceptions and views of extending their role to the examination of the newborn baby. AB - OBJECTIVES: to explore midwives' attitudes and perceptions about extending their role to the examination of the newborn baby, as well as their general perceptions and attitudes to new role developments. DESIGN: qualitative, data collected using semi-structured interviews, which were exploratory and interactive in form. SETTING: six maternity hospitals in South-east England. PARTICIPANTS: ten midwives were purposefully selected, including five trained in the examination of the newborn baby and currently conducting examinations and five who had not. Most of the midwives had been qualified for over ten years and had a wide range of clinical experience in hospital and community settings. FINDINGS: midwives identified many benefits to themselves, to their profession and to the mothers as a result of developing their role into the examination of the newborn baby. The major benefit cited was improved job satisfaction, which was directly related to their ability to give continuity and total care to mothers and babies. Midwives also perceived that undertaking the examination strengthened their position as autonomous practitioners, by enabling them to provide total care to mothers and babies who fitted their criteria of normality. Moreover, midwives thought that improvements in the overall quality of care to mothers would result from them performing the examination, including improved communication, greater continuity of care and a more holistic examination. Although midwives were concerned about possible increase in workloads and pressure to take on new roles, the examination was generally perceived as being easily incorporated into their current practice without compromising overall standards in midwifery care. Midwives expressed concern about 'extending' practice into areas that did not fit their perceptions of normality and about being 'pressurised' into taking on new roles. IMPLICATIONS FOR PRACTICE: it would appear from this study that an important consideration for midwives in their acceptance of new roles, is the relationship of that role to their position as experts in normality. Midwives in this study viewed the examination of the newborn baby as 'fitting in' with their perceptions of the core values of midwifery. PMID- 12634038 TI - Midwives' knowledge of, attitudes to and practice with baby walkers. AB - OBJECTIVE: to determine community midwives' knowledge, attitudes and practice with regard to baby walkers. DESIGN: survey, using questionnaires. SETTING: primary care, East Midlands, UK. PARTICIPANTS: Sixty-five community midwives participating in a cluster randomised controlled trial to reduce baby walker use. FINDINGS: there was a 94% response. Seventy per cent of the midwives, felt that it was part of their role to discuss baby walkers, and 62% felt that giving advice about walkers before birth acceptable. The midwives had a limited knowledge of baby walker injury risk and only one midwife had any baby walker related health education material. Three attitude scales were computed and indicated that the midwives held a negative view of baby walkers and were positive about baby walker health education, believing that parents hold a positive attitude to baby walkers. Those midwives who had used a walker with their own children were more positive about baby walkers and less positive about baby walker health education. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: midwives are willing to be involved in baby walker health education during pregnancy. However, they require evidence-based knowledge and materials rather than relying on personal experience. PMID- 12634039 TI - Self-organized criticality in ant brood tending. AB - A new stochastic lattice gas model of ant brood tending is formulated to examine the role played by repulsive ant-ant interactions in the even distribution of care among brood members. The deterministic limit of the model is known to be self-organized critical. Numerical simulations of the model show that the ant-ant repulsion facilitates an even distribution of brood care in the middle of the brood. This provides a possible explanation for the fact that ants sort their brood so that the youngest brood (which are most in need of care) are placed in the middle. Simulations show that the uniformity of brood care distribution is optimal when ants operate in a regime intermediate between completely random and completely deterministic. A certain degree of randomness helps ants to avoid becoming trapped in suboptimal configurations but does not destroy the long-range correlations that are inherent to self-organized critical systems. PMID- 12634040 TI - Self-organized criticality and emergent oscillations in models of termite architecture with crowding. AB - The termite architecture model of O'Toole et'al. (1999) is extended to incorporate arbitrary halting time-scales. It is shown that this also means that the assumption of synchronous building must be relaxed. Numerical simulations show that ordered nest architecture emerges under a wide range of time-scales but also that there is an optimal region of halting times. This optimal region is explained by the emergence of synchronized periods of termite activity. The correlation length of the building distribution is shown to diverge providing strong evidence that the model is self-organized critical. PMID- 12634042 TI - Probabilities of encounters between objects in biological systems 2: cognizer view. AB - The same event may not necessarily occur against a given cognition (action) upon repetition. The degree of certainty in which a particular event actually occurs following a current cognition is the probability of the event viewed (experienced) by the focal cognizer. This is the internal concept of probability, which is contrasted with the external probability concept, the probability viewed by the meta-observer located outside the system, expressed as the relative frequencies of events. The internal probability may be a measure of the adaptive cognitions or actions of a cognizer by which it generates or maintains a particular relation to its surroundings. In this paper, the quantitative description of encounter probabilities based on the internal concept is explored in the framework of the cognizers system model. The internal analysis derives the same formulation as that derived by the meta-observer concept of probability reported in a previous paper (Nakajima, 2001a), indicating the theoretical consistency of encounter probabilities between the analyses by the two contrasting probability concepts. The formulation suggests that a cognizer can raise (or reduce) the encounter probability with targets by a discriminative and selective response to a given situation, which describes a quantitative aspect of biological adaptation to the environment. PMID- 12634041 TI - Key-string segmentation algorithm and higher-order repeat 16mer (54 copies) in human alpha satellite DNA in chromosome 7. AB - A new key-string segmentation algorithm for identification of alpha satellite DNAs and higher-order repeat (HOR) units was introduced and exemplified. Starting with an initial key string, we determine the dominant key string and HOR. Our key string algorithm was used to scan the recent GenBank data for human alpha satellite DNA sequence AC017075.8 (193 277 bp) from the centromeric region of chromosome 7. The sequence was computationally segmented into one HOR domain (super-repeat domain) and two non-HOR domains. Dominant key-string GTTTCT provided segmentation in terms of alpha monomers. The HOR is tandemly repeated in 54 copies in the super-repeat (HOR) domain. Five insertions and three deletions in the HOR structure associated with a dominant key string were identified. Concensus HOR was constructed. Divergence of individual HOR copies from concensus amounts to 0.7% on the average, while divergence between 16 monomer variants within each HOR is on the average 20%. In the front and back domain, 199 monomer variants were identified that are not organized in HOR and diverge by 20-40%. PMID- 12634043 TI - Strategic choice handicaps when females seek high male net viability. AB - We examine a strategic-choice handicap model in which males send costly signals to advertise their quality to females. Females are concerned with the net viability of the male with whom they mate, where net viability is a function of the male's quality and signal. We identify circumstances in which a signaling equilibrium would require high-quality males to send signals so much larger than those of lower-quality males (to deter mimicry by the latter) as to yield lower net viabilities for the former. This causes females to shun males who send large signals, ensuring that there is no signaling equilibrium. PMID- 12634044 TI - Coexistence in a competitive parasitoid-host system. AB - The main objective of this work is to determine the conditions for coexistence and competitive exclusion in a discrete model for a community of three species: a stage-structured host and two competing parasitoids sharing the same host developmental stage. Coexistence of the community of the species is found to depend on the host life history parameters in the first place, and on competitive ability and parasitoid efficiency in the second place. In particular, parasitoids equilibrium densities are defined by the size of the refuge. Extinction is expected with low growth rate and with low adult survival. Host life histories are also associated with oscillations in population density, and depending on the combination of host adult survival from one generation to the next and host growth rate, the minimum of fluctuations approaches zero, implying a higher potential risk of extinction because of stochastic factors. Our results suggest that equally reduced survival of parasitoids in hosts parasitized by both species determines extinction of the parasitoid with lower population density, in contrast to the case when both parasitoids benefit with 50% of all doubly parasitized hosts, leading to the hypothesis that a community where competitors in multiparasitized hosts die, easily becomes extinct. Competitive exclusion is expected for highly asymmetric competitive interactions, independent of population densities, allowing us to hypothesize that coexistence of competitors in systems with limited resources and refuges is associated with a clearly defined competitive hierarchy. PMID- 12634045 TI - A mathematical model of single target site location by Brownian movement in subcellular compartments. AB - The location of distinct sites is mandatory for many cellular processes. In the subcompartments of the cell nucleus, only very small numbers of diffusing macromolecules and specific target sites of some types may be present. In this case, we are faced with the Brownian movement of individual macromolecules and their "random search" for single/few specific target sites, rather than bulk averaged diffusion and multiple sites. In this article, I consider the location of a distant central target site, e.g. a globular protein, by individual macromolecules executing unbiased (i.e. drift-free) random walks in a spherical compartment. For this walk-and-capture model, the closed-form analytic solution of the first passage time probability density function (p.d.f.) has been obtained as well as the first and second moment. In the limit of a large ratio of the radii of the spherical diffusion space and central target, well-known relations for the variance and the first two moments for the exponential p.d.f. were found to hold with high accuracy. These calculations reinforce earlier numerical results and Monte Carlo simulations. A major implication derivable from the model is that non-directed random movement is an effective means for locating single sites in submicron-sized compartments, even when the diffusion coefficients are comparatively small and the diffusing species are present in one copy only. These theoretical conclusions are underscored numerically for effective diffusion constants ranging from 0.5 to 10.0 microm(2) s(-1), which have been reported for a couple of nuclear proteins in their physiological environment. Spherical compartments of submicron size are, for example, the Cajal bodies (size: 0.1-1.0 microm), which are present in 1-5 copies in the cell nucleus. Within a small Cajal body of radius 0.1 microm a single diffusing protein molecule (with D=0.5 microm(2) s(-1)) would encounter a medium-sized protein of radius 2.5 nm within 1 s with a probability near certainty (p=0.98). PMID- 12634046 TI - A mechanics model for the compression of plant and vegetative tissues. AB - The mechanics analysis of plant or vegetable tissue under a compressive stress has been developed based on large deformation elasticity theory. The tissue was treated as a lattice of regular perfect three-dimensional hexagonal cells. The cell walls were assumed to be impermeable under the time-scale of the loading. The cell walls of plants and vegetables are polymeric composite materials, consisting of a relatively amorphous matrix and a highly structured network of microfibrils embedded in the cell wall matrix. The micromechanical features of the individual cells have been related to the macroscopic properties of the whole tissue. The effects of microfibrillar stiffening factors k(1) and k(2), the cell wall matrix property alpha and the initial cell expansion ratio nu(i) on the compressive behaviour of a plant or vegetable tissue have been investigated. The predicted results have also been related to some experimental evidence. PMID- 12634047 TI - Fertility assurance through extrapair fertilizations and male paternity defense. AB - Extrapair paternity has been observed in many formally monogamous species. Male pursuit of extrapair fertilizations is explained by the advantages of having offspring that receive essential paternal care from other males. Since females are capable of exercising a degree of control over the post-copulatory sperm competition, extrapair paternity cannot persist unless it confers fitness benefits on cuckolding females. Thus, extrapair paternity involves cooperation between mated females and extrapair males. On the other hand, paired males frequently exhibit strategies that minimize their loss of paternity and/or conserve paternal investment if paternity is lost. Hence, extrapair attributes of diverse species and populations reported in the literature are particular solutions of evolutionary games involving gender-specific cuckolding/anti cuckolding strategies. Here we use methods of evolutionary game theory to study the role of male paternity guarding strategies in situations where females seek extrapair fertilizations for reasons of genetic compatibility and/or in pursuit of genetic diversity for their offspring. Our results indicate that in these circumstances pursuit of extrapair fertilizations is the only evolutionary stable female strategy. Males, on the other hand, have two, mutually exclusive, evolutionary stable strategies: full time pursuit of extrapair fertilizations and a compromise strategy wherein they protect in-pair paternity during their mate's fertile periods and pursue extrapair paternity the rest of the time. The relative merits of these two strategies are determined by the efficiency of male in-pair paternity defense, breeding synchrony, fitness advantages of extrapair over in pair offspring, and the intensity of competition for extrapair fertilizations from floater males. PMID- 12634048 TI - Support vector machines for prediction of protein domain structural class. AB - The support vector machines (SVMs) method was introduced for predicting the structural class of protein domains. The results obtained through the self consistency test, jack-knife test, and independent dataset test have indicated that the current method and the elegant component-coupled algorithm developed by Chou and co-workers, if effectively complemented with each other, may become a powerful tool for predicting the structural class of protein domains. PMID- 12634050 TI - Some unusual considerations about vessel walls and wall stresses. AB - The "zero-stress state" of blood vessels is usually defined with respect to the atmospheric pressure p(a) ( approximately 750 mmHg). As a consequence, circumferential and axial wall stresses due to a positive transmural pressure can only be positive and thus, by definition, only tensile. If the zero-stress state were defined with respect to vacuum pressure (0 mmHg), the compressive stress p(a) generated by p(a) everywhere in the wall would, however, be included so that negative (=compressive) wall stresses would formally become possible. In order to examine the consequences this alternative definition would have for arteries, we have compared radial, circumferential, and axial stresses calculated "conventionally" to the values they take when the zero-stress state is defined "correctly" by reference to the vacuum pressure. It turns out that, under normal physiologic conditions, axial stress and perhaps also circumferential stress might well be compressive in many elastic and conductance arteries, contrary to the intuitive conviction of many people. Since the type of stresses a vessel wall is submitted to may be highly relevant for its structure and mechanical properties, this unconventional way of considering wall stresses may reveal unsuspected relationships between wall stresses on one side, and wall structure, vessel growth, adaptation and repair processes, atherosclerosis, angioplasty or stenting on the other side. Similar considerations might also prove useful with regard to cardiac hypertrophy. PMID- 12634049 TI - Antibody dynamics in childhood diseases: waning and boosting of immunity and the impact of vaccination. AB - The introduction of vaccination against acute diseases such as measles induced a dramatic decline in the prevalence of the disease, and a more gradual rise in the proportion of the population whose immunity is derived solely from vaccination. These two factors combine to constitute an important shift in the dynamics of immunity, especially in highly vaccinated populations. We develop a general model to describe both loss of immunity in the absence of disease, and boosting of immunity corresponding to subclinical infection in individuals whose immunity has waned. We consider the interaction between infection and immunity and identify the key parameters that determine the eradication threshold. We explore the dynamics in the years following the introduction of vaccination using a stochastic version of the model, and consider the effect of different assumptions concerning the nature of immunity. A comparison of the model results with recently published data suggests that heterogeneity in host immune response is an important feature of the antibody dynamics. PMID- 12634051 TI - The thrifty phenotype hypothesis: thrifty offspring or thrifty mother? AB - Medical research is increasingly focusing on the contribution of nutritional programming to disease in later life. Programming is a process whereby a stimulus during a critical window of time permanently affects subsequent structure, function or developmental schedule of the organism. The thrifty phenotype hypothesis is widely used to interpret such studies, with early growth restriction seen as adaptation to environmental deprivation. However, such permanent adjustment is less beneficial than maintaining flexibility so as to recover from early growth deficits if the environment improves. Thus, the existing thrifty phenotype hypothesis fails to explain why plasticity is lost so early in development in species with extended growth. One explanation is that the developing organism simply cannot maintain phenotypic plasticity throughout the period of organ growth. This article adds a life history perspective, arguing that programming of the offspring may in some species benefit maternal fitness more than it does that of individual offspring. Closing the critical window early in development allows the preservation of maternal strategy in offspring phenotype, which in humans benefits the mother by constraining offspring demand after weaning. The offspring gains by being buffered against environmental fluctuations during the most sensitive period of development, allowing coherent adaptation of organ growth to the state of the environment. The critical window is predicted to close when offspring physiology becomes independent of maternal physiology, the timing of which depends on offspring trait. Because placental nutrition and lactation buffer against short-term environmental fluctuations, maternal strategy is predicted to derive from long-term experience, encapsulated in maternal size and nutritional status. Such an approach implies that public health programmes for improving birth weight may be more effective if they target maternal development rather than nutrition during pregnancy. Equally, aggressive nutritional management of infants born small or pre-term may induce the very environmental fluctuations that are naturally softened by maternal nutrition. PMID- 12634052 TI - Specific inhibition of the translocation of a subset of Escherichia coli TAT substrates by the TorA signal peptide. AB - The SufI protein and the trimethylamine N-oxide reductase (TorA) are the two best characterized prototype proteins exported by the Escherichia coli TAT system. Whereas SufI does not contain cofactors, TorA is a molybdo-enzyme and the acquisition of the molybdo-cofactor is a prerequisite for its translocation. The overproduction of each protein leads to the saturation of its translocation, but it was unknown if the overproduction of one substrate could saturate the TAT apparatus and block thus the translocation of other TAT substrates. Here, we showed that the overproduction of SufI saturated only its own translocation, but had no effect of the translocation of TorA and other TAT substrate analyzed. To dissect the saturation mechanism of TorA translocation, we shortened by about one third of the TorA protein and removed nine consensus molybdo-cofactor-binding ligands. Like SufI, the truncated TorA (TorA502) did not contain cofactor and would not compete with the full length TorA for molybdo-cofactor acquisition. The overproduction of TorA502 completely inhibited the export of the full length TorA and dimethyl sulfoxide (DMSO) reductase, but had no effect on the translocation of SufI, nitrate-induced formate dehydrogenase and hydrogenase-2. Importantly, deletion of the twin-arginine signal peptide of TorA502 abolished the inhibitory effect. Moreover, the overproduction of the TorA signal peptide fused to the green fluorescence protein (GFP) was sufficient to block the TorA translocation. These results demonstrated that the twin-arginine signal peptide of the TorA protein specifically inhibits the translocation of a subset of TAT substrates, probably at the step of their targeting to the TAT apparatus. PMID- 12634053 TI - Kinetic regulation of single DNA molecule denaturation by T4 gene 32 protein structural domains. AB - Bacteriophage T4 gene 32 protein (gp32) specifically binds single-stranded DNA, a property essential for its role in DNA replication, recombination, and repair. Although on a thermodynamic basis, single-stranded DNA binding proteins should lower the thermal melting temperature of double-stranded DNA (dsDNA), gp32 does not. Using single molecule force spectroscopy, we show for the first time that gp32 is capable of slowly destabilizing natural dsDNA. Direct measurements of single DNA molecule denaturation and renaturation kinetics in the presence of gp32 and its proteolytic fragments reveal three types of kinetic behavior, attributable to specific protein structural domains, which regulate gp32's helix destabilizing capabilities. Whereas the full-length protein exhibits very slow denaturation kinetics, a truncate lacking the acidic C-domain exhibits much faster kinetics. This may reflect a steric blockage of the DNA binding site and/or a conformational change associated with this domain. Additional removal of the N-domain, which is needed for binding cooperativity, further increases the DNA denaturation rate, suggesting that both of these domains are critical to the regulation of gp32's helix-destabilization capabilities. This regulation is potentially biologically significant because uncontrolled helix-destabilization would be lethal to the cell. We also obtain equilibrium measurements of the helix coil transition free energy in the presence of these proteins for the first time. PMID- 12634054 TI - Subunit assembly for DNA cleavage by restriction endonuclease SgrAI. AB - The SgrAI endonuclease usually cleaves DNA with two recognition sites more rapidly than DNA with one site, often converting the former directly to the products cut at both sites. In this respect, SgrAI acts like the tetrameric restriction enzymes that bind two copies of their target sites before cleaving both sites concertedly. However, by analytical ultracentrifugation, SgrAI is a dimer in solution though it aggregates to high molecular mass species when bound to its specific DNA sequence. Its reaction kinetics indicate that it uses different mechanisms to cleave DNA with one and with two SgrAI sites. It cleaves the one-site DNA in the style of a dimeric restriction enzyme acting at an individual site, mediating neither interactions in trans, as seen with the tetrameric enzymes, nor subunit associations, as seen with the monomeric enzymes. In contrast, its optimal reaction on DNA with two sites involves an association of protein subunits: two dimers bound to sites in cis may associate to form a tetramer that has enhanced activity, which then cleaves both sites concurrently. The mode of action of SgrAI differs from all restriction enzymes characterised previously, so this study extends the range of mechanisms known for restriction endonucleases. PMID- 12634055 TI - How does an enzyme evolved in vitro compare to naturally occurring homologs possessing the targeted function? Tyrosine aminotransferase from aspartate aminotransferase. AB - Aspartate aminotransferase (AATase) and tyrosine aminotransferase (TATase) are Escherichia coli paralogs that share 43% sequence identity. A plausible model posits that TATase arose from a duplication of an ancestral AATase-like enzyme. Directed evolution of AATase to an enzyme having TATase activity was undertaken in order to compare the evolved AATase variants with homologous TATases. Eight rounds of DNA shuffling and in vivo selection followed by a backcross with WT AATase produced enzymes that exhibited 100-270-fold increases in k(cat)/K(m)(Phe) and had as much as 11% of the tyrosine aminotransferase activity of WT E.coli TATase. Amino acid substitutions in 11 clones from rounds 7 and 8 were compared with conserved residues in AATases and TATases. The findings are conveniently and compactly illustrated by the use of Venn diagrams and set theory notation. A statistically significant (0.001or=75% identical) in AATases and variable (<75% identical) in TATases. Very few mutations occur in the intersection (set AAT intersection TAT) of amino acid residues that are conserved in both enzyme types. Seven mutations from set AAT-TAT were combined by site directed mutagenesis to give a construct that is 60% as active as the best round 8 enzyme, which has 13 amino acid replacements. The Venn diagrams may provide a generally useful tool to highlight the most important specificity determinants for rational redesign. Amino acid replacements were mapped onto the crystal structure of a hydrocinnamate complex of a designed TATase. Five of the seven positions most frequently substituted in the evolved clones are within 15 A of the phenyl side-chain, but only six of the 48 positions that were mutated once or twice are within that radius. Context dependence, neutral mutations, different selective pressures, and stochastic components provide explanations for the observation that many of the substitutions found in the directly evolved enzymes differ from the corresponding amino acids found in the modern natural TATases. PMID- 12634056 TI - A kinase anchoring protein (AKAP) interaction and dimerization of the RIalpha and RIbeta regulatory subunits of protein kinase a in vivo by the yeast two hybrid system. AB - Protein kinase A (PKA) regulatory (R) subunits dimerize through an N-terminal motif. Such dimerization is necessary for binding to PKA anchoring proteins (AKAPs) and targeting of PKA to its site of action. In the present study, we used the yeast two-hybrid system as an in vivo bio-reporter assay and analyzed the formation of homo- and heterodimeric complexes of RIalpha and RIbeta as well as AKAP binding of RI dimers. Native polyacrylamide gel electrophoresis (PAGE) of yeast extracts confirmed the two-hybrid data. Both RIalpha- and RIbeta homodimers as well as an RIalpha:RIbeta heterodimer were observed. Single, double and one triple mutation were introduced into the RIalpha and RIbeta subunits and dimerization properties of the mutants were analyzed. Consistent with previous reports, RIalpha(C37H) dimerized, although the disulfide bridges were disrupted, whereas the additional mutation of F47 or F52 abolished the dimerization. Corresponding mutations (C38H, F48A, F53A) in RIbeta were not sufficient to abolish the RIbeta dimerization, indicating that additional or other amino acids are important. RIalpha:RIbeta heterodimers of the mutants were formed at intermediate stringency. Analysis of ternary complexes by the yeast two-hybrid system revealed that RIalpha and RIbeta homodimers as well as an RIalpha:RIbeta heterodimer and several of the mutants were able to bind to the R-binding domain of AKAP149/D-AKAP1. Furthermore, an RIbeta:AKAP149 complex was identified following introduction of RIbeta into HEK293 cells. Importantly, RIbeta revealed AKAP binding properties similar to those of RIalpha, indicating that RIbeta holoenzymes may be anchored. PMID- 12634057 TI - Motions and negative cooperativity between p97 domains revealed by cryo-electron microscopy and quantised elastic deformational model. AB - p97, a Mg-ATPase belonging to the AAA (ATPase associated with various cellular activities) super family of proteins, has been proposed to function in two distinct cellular pathways, namely homotypic membrane fusion and ubiquitin protein degradation by utilizing differing adaptor complexes. We present the cryo electron microscopy three-dimensional reconstruction of endogenous p97 in an AMP PNP bound state at 24 A resolution. It reveals clear nucleotide-dependent differences when compared to our previously published "p97-ADP" reconstruction, including a striking rearrangement of N domains and a positional change of the two ATPase domains, D1 and D2, with respect to each other. The docking of the X ray structure of N-D1 domains in an ADP bound state indicates that an upward repositioning of N domain is necessary to accommodate the cryo-EM map of "p97-AMP PNP", suggesting a change in the orientation of N domains upon nucleotide hydrolysis. Furthermore, computational analysis of the deformational motions of p97, performed on the cryo-EM density map and the atomic structure of the N-D1 domains independently, shows the existence of a negative cooperativity between the D1 and D2 rings and the flexibility of the N domains. Together these results allow the identification of functionally important features that offer molecular insights into the dynamics of the proposed p97 chaperone function. PMID- 12634058 TI - The structure of bovine lysosomal alpha-mannosidase suggests a novel mechanism for low-pH activation. AB - Lysosomal alpha-mannosidase (LAM: EC 3.2.1.24) belongs to the sequence-based glycoside hydrolase family 38 (GH38). Two other mammalian GH38 members, Golgi alpha-mannosidase II (GIIAM) and cytosolic alpha-mannosidase, are expressed in all tissues. In humans, cattle, cat and guinea pig, lack of lysosomal alpha mannosidase activity causes the autosomal recessive disease alpha-mannosidosis. Here, we describe the three-dimensional structure of bovine lysosomal alpha mannosidase (bLAM) at 2.7A resolution and confirm the solution state dimer by electron microscopy. We present the first structure of a mammalian GH38 enzyme that offers indications for the signal areas for mannose phosphorylation, suggests a previously undetected mechanism of low-pH activation and provides a template for further biochemical studies of the family 38 glycoside hydrolases as well as lysosomal transport. Furthermore, it provides a basis for understanding the human form of alpha-mannosidosis at the atomic level. The atomic coordinates and structure factors have been deposited in the Protein Data Bank (accession codes 1o7d and r1o7dsf). PMID- 12634059 TI - Bivalent binding of IgA1 to FcalphaRI suggests a mechanism for cytokine activation of IgA phagocytosis. AB - FcalphaRI, the receptor specific for the Fc region of immunoglobulin A (IgA), is responsible for IgA-mediated phagocytosis, oxidative burst, and antibody dependent cellular cytotoxicity. Using the techniques of analytical ultracentrifugation and equilibrium gel-filtration, we show that two FcalphaRI molecules bind to a single Fcalpha homodimer. Surface plasmon resonance studies confirm the 2:1 stoichiometry of binding, with equilibrium dissociation constants of 176 nM and 431 nM for the first and second binding events, respectively. The binding affinity decreases at acidic pH in a manner consistent with protonation of a single histidine residue in the binding site. A thermodynamic analysis indicates that the histidine residue does not participate in a salt-bridge in the complex; in fact, less than 10% of the free energy of binding was contributed by electrostatic interactions. The bivalent, pH-dependent interaction between FcalphaRI and IgA has important implications for cytokine-dependent phagocytosis of IgA and the FcalphaRI-mediated degradation or recycling of IgA. PMID- 12634060 TI - Structure and ligand binding of carbohydrate-binding module CsCBM6-3 reveals similarities with fucose-specific lectins and "galactose-binding" domains. AB - Carbohydrate-binding polypeptides, including carbohydrate-binding modules (CBMs) from polysaccharidases, and lectins, are widespread in nature. Whilst CBMs are classically considered distinct from lectins, in that they are found appended to polysaccharide-degrading enzymes, this distinction is blurring. The crystal structure of CsCBM6-3, a "sequence-family 6" CBM in a xylanase from Clostridium stercorarium, at 2.3 A reveals a similar, all beta-sheet fold to that from MvX56, a module found in a family 33 glycoside hydrolase sialidase from Micromonospora viridifaciens, and the lectin AAA from Anguilla anguilla. Sequence analysis leads to the classification of MvX56 and AAA into a family distinct from that containing CsCBM6-3. Whilst these polypeptides are similar in structure they have quite different carbohydrate-binding specificities. AAA is known to bind fucose; CsCBM6-3 binds cellulose, xylan and other beta-glucans. Here we demonstrate that MvX56 binds galactose, lactose and sialic acid. Crystal structures of CsCBM6-3 in complex with xylotriose, cellobiose, and laminaribiose, 2.0 A, 1.35 A, and 1.0 A resolution, respectively, reveal that the binding site of CsCBM6-3 resides on the same polypeptide face as for MvX56 and AAA. Subtle differences in the ligand binding surface give rise to the different specificities and biological activities, further blurring the distinction between classical lectins and CBMs. PMID- 12634061 TI - Assembly of protein subunits within the stromal ridge of photosystem I. Structural changes between unbound and sequentially PS I-bound polypeptides and correlated changes of the magnetic properties of the terminal iron sulfur clusters. AB - The X-ray structure of Photosystem I (PS I) from Synechococcus elongatus was recently solved at 2.5A resolution (PDB entry 1JB0). It provides a structural model for the stromal subunits PsaC, PsaD and PsaE, which comprise the "stromal ridge" of PS I. In a separate set of studies the three-dimensional solution structures of the unbound, recombinant PsaC (PDB entry 1K0T) and PsaE (PDB entries 1PSF, 1QP2 and 1GXI) subunits were solved by NMR. The PsaC subunit of PS I is a small (9.3 kDa) protein that harbors binding sites for two [4Fe-4S] clusters F(A) and F(B), which are the terminal electron acceptors in PS I. Comparison of the PsaC structure in solution with that in the X-ray structure of PS I reveals significant differences between them which are summarized and evaluated here. Changes in the magnetic properties of [4Fe-4S] centers F(A) and F(B) are related to changes in the protein structure of PsaC, and they are further influenced by the presence of PsaD. Based on experimental evidence, three assembly stages are analyzed: PsaC(free), PsaC(only), PsaC(PS I). Unbound, recombinant PsaD, studied by NMR, has only a few elements of secondary structure and no stable three-dimensional structure in solution. When PsaD is bound in PS I, it has a well-defined three-dimensional structure. For PsaE the three dimensional structure is very similar in solution and in the PS I-bound form, with the exception of two loop regions. We suggest that the changes in the structures of PsaC and PsaD are caused by the sequential formation of multiple networks of contacts between the polypeptides of the stromal ridge and between those polypeptides and the PsaA/PsaB core polypeptides. The three-dimensional structure of the C(2)-symmetric F(X)-binding loops on PsaA and PsaB were also analyzed and found to be significantly different from the binding sites of other proteins that contain interpolypeptide [4Fe-4S] clusters. The aim of this work is to relate contact information to structural changes in the proteins and to propose a model for the assembly of the stromal ridge of PS I based on this analysis. PMID- 12634062 TI - Reversible amyloid formation by the p53 tetramerization domain and a cancer associated mutant. AB - The tetramerization domain for wild-type p53 (p53tet-wt) and a p53 mutant, R337H (p53tet-R337H), associated with adrenocortical carcinoma (ACC) in children, can be converted from the soluble native state to amyloid-like fibrils under certain conditions. Circular dichroism, Fourier transform infrared spectroscopy and staining with Congo red and thioflavin T showed that p53tet-wt and p53tet-R337H adopt an alternative beta-sheet conformation (p53tet-wt-beta and p53tet-R337H beta, respectively), characteristic of amyloid-like fibrils, when incubated at pH 4.0 and elevated temperatures. Electron micrographs showed that the alternative conformations for p53tet-wt (p53tet-wt-beta) and p53tet-R337H (p53tet-R337H-beta) were supramolecular structures best described as "molecular ribbons". FT-IR analysis demonstrated that the mechanism of amyloid-like fibril formation involved unfolding of the p53tet-wt beta-strands, followed by unfolding of the alpha-helices, followed finally by formation of beta-strand-containing structures that other methods showed were amyloid-like ribbons. The mutant, p53tet-R337H, had a significantly higher propensity to form amyloid-like fibrils. Both p53tet wt (pH 4.0) and p53tet-R337H (pH 4.0 and 5.0), when incubated at room temperature (22 degrees C) for one month, were converted to molecular ribbons. In addition, p53tet-R337H, and not p53tet-wt, readily formed ribbons at pH 4.0 and 37 degrees C over 20 hours. Interestingly, unlike other amyloid-forming proteins, p53tet-wt beta and p53tet-R337H-beta disassembled and refolded to the native tetramer conformation when the solution pH was raised from 4.0 to 8.5. Although fibril formation at pH 4.0 was concentration and temperature-dependent, fibril disassembly at pH 8.5 was independent of both. Finally, we propose that the significantly higher propensity of the mutant to form ribbons, compared to the wild-type, may provide a possible mechanism for the observed nuclear accumulation of p53 in ACC cells and other cancerous cells. PMID- 12634063 TI - Ab initio folding simulation of the Trp-cage mini-protein approaches NMR resolution. AB - Here, we report a 100 ns molecular dynamics simulation of the folding process of a recently designed autonomous-folding mini-protein designated as tc5b with a new AMBER force field parameter set developed based on condensed-phase quantum mechanical calculations and a Generalized Born continuum solvent model. Starting from its fully extended conformation, our simulation has produced a final structure resembling that of NMR native structure to within 1A main-chain root mean square deviation. Remarkably, the simulated structure stayed in the native state for most part of the simulation after it reached the state. Of greater significance is that our simulation has not only reached the correct main-chain conformation, but also a very high degree of accuracy in side-chain packing conformation. This feat has traditionally been a challenge for ab initio simulation studies. In addition to characterization of the trajectory, comparison of our results to experimental data is also presented. Analysis of the trajectory suggests that the rate-limiting step of folding of this mini-protein is the packing of the Trp side-chain. PMID- 12634064 TI - Increased backbone mobility in beta-barrel enhances entropy gain driving binding of N-TIMP-1 to MMP-3. AB - The high-affinity inhibition of stromelysin 1 (MMP-3) by tissue inhibitor of metalloproteinases 1 (TIMP-1) helps control tissue remodeling and tumor development. The interaction of N-TIMP-1 with the catalytic domain of MMP-3 has been investigated by titration calorimetry and 15N NMR. Their unfavorable enthalpy of binding of +6.5 kcal mol(-1) is unusual among protein-protein associations, deviates from structure-based prediction, and is compensated by a net entropy increase providing at least 18 kcal mol(-1) of favorable free energy of binding at a 1M reference state. The small heat capacity of binding agrees well with the heat capacity predicted from 65% of the surface buried on binding being polar, and suggests that the hydrophobic effect can account for only part of the entropy of binding. Using NMR, binding-induced changes in the backbone of N-TIMP-1 were checked as one possible source of conformational entropy changes. MMP binding slightly increases rigidity in some contact sites in TIMP-1 but increases mobility remotely in the otherwise rigid beta-barrel core of N-TIMP-1, increasing 15N relaxation evidence of pico- to nanosecond and micro- to millisecond fluctuations of beta-strands A-F. Residual dipolar couplings suggest dynamic deviations from X-ray coordinates of the complex. These suggest that the beta-barrel has small backbone conformational fluctuations, while segments of strands betaB, betaE and betaF might experience fluctuations only in their backbone environment. This is a distinctive example of affinity between two well structured proteins being enhanced by increased conformational entropy in the reservoir of a folding core. PMID- 12634065 TI - Reliability measures for membrane protein topology prediction algorithms. AB - We have developed reliability scores for five widely used membrane protein topology prediction methods, and have applied them both on a test set of 92 bacterial plasma membrane proteins with experimentally determined topologies and on all predicted helix bundle membrane proteins in three fully sequenced genomes: Escherichia coli, Saccharomyces cerevisiae and Caenorhabditis elegans. We show that the reliability scores work well for the TMHMM and MEMSAT methods, and that they allow the probability that the predicted topology is correct to be estimated for any protein. We further show that the available test set is biased towards high-scoring proteins when compared to the genome-wide data sets, and provide estimates for the expected prediction accuracy of TMHMM across the three genomes. Finally, we show that the performance of TMHMM is considerably better when limited experimental information (such as the in/out location of a protein's C terminus) is available, and estimate that at least ten percentage points in overall accuracy in whole-genome predictions can be gained in this way. PMID- 12634066 TI - Protein dynamics in a family of laboratory evolved thermophilic enzymes. AB - Molecular dynamics simulations were employed to study how protein solution structure and dynamics are affected by adaptation to high temperature. Simulations were carried out on a para-nitrobenzyl esterase (484 residues) and two thermostable variants that were generated by laboratory evolution. Although these variants display much higher melting temperatures than wild-type (up to 18 degrees C higher) they are both >97% identical in sequence to the wild-type. In simulations at 300 K the thermostable variants remain closer to their crystal structures than wild-type. However, they also display increased fluctuations about their time-averaged structures. Additionally, both variants show a small but significant increase in radius of gyration relative to wild-type. The vibrational density of states was calculated for each of the esterases. While the density of states profiles are similar overall, both thermostable mutants show increased populations of the very lowest frequency modes (<10 cm(-1)), with the more stable mutant showing the larger increase. This indicates that the thermally stable variants experience increased concerted motions relative to wild-type. Taken together, these data suggest that adaptation for high temperature stability has resulted in a restriction of large deviations from the native state and a corresponding increase in smaller scale fluctuations about the native state. These fluctuations contribute to entropy and hence to the stability of the native state. The largest changes in localized dynamics occur in surface loops, while other regions, particularly the active site residues, remain essentially unchanged. Several mutations, most notably L313F and H322Y in variant 8G8, are in the region showing the largest increase in fluctuations, suggesting that these mutations confer more flexibility to the loops. As a validation of our simulations, the fluctuations of Trp102 were examined in detail, and compared with Trp102 phosphorescence lifetimes that were previously measured. Consistent with expectations from the theory of phosphorescence, an inverse correlation between out-of-plane fluctuations on the picosecond time scale and phosphorescence lifetime was observed. PMID- 12634067 TI - Early management of suspected bacterial meningitis and meningococcal septicaemia in adults. PMID- 12634068 TI - Use of pneumococcal polysaccharide vaccines: no simple answers. AB - Control of pneumococcal disease by vaccination with capsular polysaccharide has been investigated and promoted with variable enthusiasm for the past 90 years. Despite this the true value of the 23-valent pneumococcal polysaccharide vaccine remains unclear. In particular the ability of this vaccine to provide durable protection in those who require it most is in greatest doubt. With the introduction of the protein-conjugated pneumococcal polysaccharide vaccines new opportunities for prevention exist, but it will be import to rigorously assess their role in at-risk groups to avoid the uncertainty that surrounds the older simple polysaccharide products. PMID- 12634069 TI - HIV-associated lipodystrophy syndrome: assessment and management. PMID- 12634071 TI - What can analysis of calls to NHS direct tell us about the epidemiology of gastrointestinal infections in the community? AB - OBJECTIVES: Most gastrointestinal (GI) illness within the UK goes undetected by routine surveillance. A national telephone helpline for health advice (NHS Direct) offers a new source of data on GI infection. We aim to describe NHS Direct calls suggestive of GI infection and the outcome of these calls. METHODS: Details of over 150000 telephone calls were collected from NHS Direct over a 6 month period. Calls about 'diarrhoea', 'vomiting' or 'food poisoning' were defined as GI calls and described according to the age of the patient and call outcome. RESULTS: Gastrointestinal calls accounted for 10.3% of total calls ('diarrhoea'=4.9%, 'vomiting'=5.1%). GI calls as a proportion of total calls were significantly high in children under 1 year (23.5%) and aged 1-4 years (21.5%). Call outcomes which resulted in further NHS care being recommended accounted for 72.3% of total calls and 54.5% of GI calls. CONCLUSIONS: A high proportion of NHS Direct calls were about GI symptoms especially for children under 5 years. When compared with all NHS Direct calls, GI calls were less likely to result in further NHS care being recommended. Analysis of NHS Direct calls provides further insight into GI infection in the community. PMID- 12634070 TI - A UK multicentre study of the antimicrobial susceptibility of bacterial pathogens causing urinary tract infection. AB - OBJECTIVES: To determine the prevalence of resistance amongst urinary tract pathogens against antimicrobials used to treat urinary tract infections (UTIs) in the UK to provide data to help direct empirical therapy. METHOD: During 1999 2000, a total of 1291 bacterial isolates causing UTI were collected from 8 centres in the UK. Isolates were cultured from patients with (1). community acquired UTI in those less than 65 years old (397), (2). hospital-acquired UTI other than those admitted with pyelonephritis (394), (3). pyelonephritis (108) and (4). community-acquired UTI in those greater than 65 years old (392). After re-identification, MICs for a range of antimicrobials were determined and interpreted using NCCLS procedures and interpretive guidelines. RESULTS: Escherichia coli was the predominant pathogen in all categories but the total percentage for each category varied (56.3-77.3%). The next three pathogens of importance were Enterococcus faecalis, Klebsiella pneumoniae and Proteus mirabilis which varied in prevalence slightly from category to category. The activity of amoxycillin against E. coli (51.3% susceptible) was greatly reduced as a result of beta-lactamase production and only partially restored by the addition of clavulanic acid (78.8% susceptible). Cefuroxime was very active against E. coli using parenteral form breakpoints (97.1% susceptible) but less so using oral form breakpoints (68.6% susceptible). Cefuroxime was inactive against Enterococcus spp. and Pseudomonas spp. Nitrofurantoin was very active against isolates of E. coli (96.3% susceptible) and E. faecalis but not against K. pneumoniae, P. mirabilis or Pseudomonas aeruginosa. Overall susceptibility to trimethoprim ranged from 58.1% to 84.5% for the most prevalent pathogens. Ciprofloxacin was highly active against the UTI pathogens examined in this study with susceptibilities of between 88.6% and 97.7% for the most prevalent pathogens (E. coli, n=864, 97.7% susceptible) and was the only oral agent tested with activity against Pseudomonas spp. CONCLUSION: These data provide much needed information on the prevalence of antimicrobial resistance amongst pathogens currently causing UTI in the UK. PMID- 12634072 TI - Soft tissue infections of the extremities in an orthopaedic centre in the UK. AB - OBJECTIVES: To determine the spectrum, and clinical impact of acute extremity soft tissue infections, encountered in the Orthopaedic service of an inner city hospital in UK. METHODS: Patients requiring admission for an acute limb soft tissue infection to the Orthopaedic unit of the Manchester Royal Infirmary, UK, between July 1996 and 2001 were identified from our database. Infections involving the groin and axilla, those developing within 30 days of a surgical procedure, and patients with chronic soft tissue ulcers or infections were not considered. RESULTS: Of 142 infections the majority were cellulitis (50%) and superficial abscesses (34.5%). Most were secondary to trauma (31.6%), human or animal bites (20%) and intravenous drug abuse (17.6%). Although most patients were young and otherwise healthy, ten developed significant complications: myonecrosis requiring below knee amputation (1), acute carpal tunnel syndrome (1), osteomyelitis (6), extensive skin loss requiring reconstruction (1), deep vein thrombosis (1). Seven hundred and eighty four hospital inpatient days and 143 operative interventions were devoted to these patients. The estimated cost for each episode of soft tissue infection was pound 1011. In 25% of cases earlier referral to a surgical service would have been more appropriate. CONCLUSIONS: Soft tissue infections of the extremities confer significant morbidity and impose an important burden on medical resources. PMID- 12634073 TI - Monocytic activation of protein tyrosine kinase, protein kinase A and protein kinase C induced by porins isolated from Salmonella enterica serovar Typhimurium. AB - OBJECTIVES: In the present study a monocytic cell line, U937, was used to investigate the possible involvement of protein tyrosine kinases (NT-PTKs), protein kinase A (PKA) and protein kinase C (PKC) in cell signaling pathways following Salmonella enterica serovar Typhimurium porin stimulation. METHODS: Different concentrations of porins and lipopolysaccharide (LPS) were analysed to evaluate changes in PTK activity by a non radioactive tyrosine kinase assay and in PKA and PKC phosphorylation by Western blotting analysis. The inhibitors of PTK, PKA and PKC activation used, were: 3,4-dihydroxybenzylidene-malononitrile (tyrphostin 23), inhibitor of epidermal growth factor (EGF) receptor tyrosine kinase activity; dihychloride (H-89), a selective inhibitor of PKA which is useful to discriminate between the effects of PKC and PKA; diacylglycerol kinase inhibitor II (R59949), which is useful for elucidating roles of PKC; calphostin C, a specific inhibitor of PKC. RESULTS: Porins of the outer membrane of the ST were isolated to be used as a stimulus in the performed experiments. Following porin treatment, a dose-dependent increase in PTK, PKA and PKC activation was observed. U937 monocytes pretreated with inhibitors induced an evident decrease in PTK activity and PKA and PKC phosphorylation pattern in porin stimulated monocytes. CONCLUSIONS: Our data support the important role played by NT-PTK, PKA and PKC in transducing the activating signal in macrophages stimulated with porins through the activation of the mitogen-activated protein kinase (MAPK) pathway that participate in the regulation of gene expression. PMID- 12634074 TI - Chlamydia pneumoniae survival in macrophages is regulated by free Ca2+ dependent reactive nitrogen and oxygen species. AB - OBJECTIVES: Despite an efficient macrophage immune capability, Chlamydia pneumoniae infects host cells and causes chronic diseases. To gain better insights into C. pneumoniae survival mechanisms in macrophages, its growth in regular RAW-264.7 cells (nitric oxide sufficient NO (+)) and RAW-264.7 cells (nitric oxide insufficient NO (-)) were studied. METHODS: Role of Ca(2+), NO and reactive oxygen species (ROS) during C. pneumoniae infection in macrophages were determined. RESULTS: RAW-264.7 NO (-) cells supported significantly Chlamydia growth, showing an upregulation of ROS, superoxide dismutase (SOD) and catalase activities as compared with RAW-264.7 NO (+) cell. Ascorbic acid, inducible nitric oxide synthase inhibitor and glutathione significantly prompted Chlamydia inclusion formation. Cytosolic Ca(2+) had regulatory effect on organism growth, NO generation, SOD and catalase activities in both cell types. CONCLUSIONS: These findings suggest that minimal Ca(2+) signaling in macrophages at early stages of infection, NO and ROS release have modulatory effects onC. pneumoniae survival, onset of persistence and chronicity, processes which are needed for the initiation of diseases in which C. pneumoniae has been implicated as a possible etiologic agent. PMID- 12634075 TI - Nosocomial influenza infection among post-influenza-vaccinated patients with severe pulmonary diseases. AB - OBJECTIVES: To report an outbreak of nosocomial influenza in thirteen out of twenty-two admitted patients suffering from severe lung emphysema. METHODS: Acute phase and convalescent serum samples of nine patients were collected. An antihaemagglutinin assay was performed to detect a rise in antibodies against influenza A virus. Further information about vaccination history of the patients and healthcare workers was included. RESULTS: The majority of these twenty-two patients was vaccinated with a trivalent vaccine six months earlier. The immunological response showed that the influenza A (H3N2) strain which caused these infections is similar to the vaccine strain A/Sydney/5/97. CONCLUSIONS: The staff of our institute which was not systematically vaccinated may have been the source of infection. The time elapsed between the vaccination and the infection is the probable explanation of this event. PMID- 12634076 TI - Acute haemorrhagic leukoencephalopathy: two case reports and review of the literature. AB - Acute haemorrhagic leukoencephalopathy is a rare, rapidly progressive disease of cerebral white matter associated with high fatality. The few known patients surviving have all been treated with early instigation of immuno-modulatory therapy. Here we describe two cases of this disease and underscore the importance of considering this diagnosis in the differential for any individual presenting with progressive neurological signs, an antecedent flu-like illness, and early changes on CT showing widespread disease of white matter. PMID- 12634077 TI - SPECT scans for monitoring response to pleconaril therapy in chronic enteroviral meningoencephalitis. AB - Chronic enteroviral meningoencephalitis (CEMA) is a rare complication of immunodeficient individuals and may present as insidious intellectual deterioration. Diagnosis requires isolation or PCR identification of enterovirus from the CSF. Pleconaril, a novel anti-picornaviral compound is available on a compassionate release basis to treat patients with potentially life threatening enteroviral infection. Non-invasive neuroimaging is an important new technique for both the diagnosis of encephalitis and as an objective assessment of response to treatment. We report two immunodeficient patients, one with common variable immunodeficiency and one with HIV, with an insidious presentation of CEMA. In both patients, perfusion single photon emission tomography scans were effective in monitoring treatment, correlating with clinical and virological response to pleconaril. PMID- 12634078 TI - Cytomegalovirus infection and thrombosis: a causative association? AB - Venous thrombosis is a well-organised complication of hospitalisation and has often been reported in the context of HIV infection. Less frequently cytomegalovirus (CMV) has been implicated in the development of thrombosis, although usually in the context of immunosuppression. We report the case of a young, immunocompetent individual who developed pulmonary emboli as a complication of CMV infection and describe the three proposed mechanisms by which this is thought to occur. The probable mechanism in our patient is discussed. PMID- 12634079 TI - Anterior chest wall abscess caused by Salmonella enteritidis in a healthy adult. AB - We report a case of anterior chest wall abscess in an immunocompetent adult by Salmonella enteritidis, whose food was contaminated by bird droppings. The patient did not have any gastrointestinal symptoms. Surgical excision followed by antibiotics (cefuroxime and ciprofloxacin) successfully treated the condition. To our knowledge, this is the first reported case of anterior chest wall abscess caused by S. enteritidis in an immunocompetent adult without any preceding gastrointestinal symptoms. We feel that the contamination of his food with the bird droppings was a risk factor. PMID- 12634080 TI - Distribution of HBV DNA according to age group in the Diyarbakir region (southeast Turkey). PMID- 12634081 TI - Notice on guidelines for general practitioners: for laboratory investigation of infections and antibiotic usage. PMID- 12634083 TI - Sheep scab: the disease, pathogenesis and control. PMID- 12634084 TI - Susceptibility of three genetic lines of chicks to infection with a nephropathogenic T strain of avian infectious bronchitis virus. AB - Mortality rates were compared in three genetic lines of specific pathogen-free chicks inoculated with one of two doses of a nephropathogenic strain of avian infectious bronchitis (IB) virus. The mortality rates were influenced primarily by the chick strain, but also by age and dose of virus. Chicks of the inbred S line were highly susceptible. After inoculation with a low dose of virus at 2 and 4 weeks of age, mortality was 90 and 45%, respectively. Chicks of the HWL non inbred line were also susceptible, with mortality rates after inoculation at 2 and 4 weeks of age of 70 and 25%, respectively. Chicks of the inbred W line were resistant and non-significant mortality of 10% occurred only in 2-week-old chicks inoculated with a high dose of virus. Viral distribution in tissues of susceptible S and resistant W chicks did not differ, and virus was present in the trachea, lung and kidney of chicks from both lines throughout the acute phase (between days 3 and 7) of infection. Viral titres in the trachea and kidney in susceptible S chicks were slightly but not significantly higher than in the other chicks during the acute phase of infection. Histopathological assessment indicated an earlier onset of a regenerative phase in the trachea of W chicks than in S chicks. S chicks, in contrast to W chicks, showed no signs of renal regeneration. Additionally, the kidneys of S chicks differed from those of W chicks in showing more severe nephritis, more tubular necrosis and less heterophil infiltration and lymphocytic response throughout the acute phase of infection. The results indicate that chicken lines may differ greatly in their susceptibility to fatal IB nephritis and that resistance is likely to be under the control of immune responses to viral infection. PMID- 12634085 TI - Uterine oestrogen and progesterone receptor expression in experimental pyometra in the bitch. AB - Pyometra was induced in five bitches by the intraluminal inoculation of a ligated uterine horn in metoestrus with an Escherichia coli suspension, the other horn serving as an uninoculated control. Histologically, the inoculated horns resembled those with naturally occurring pyometra, while the uninoculated horns had an inactive appearance instead of the normal metoestrus appearance. Immunohistochemically, the expression of sex hormone receptors in the inoculated horns corresponded with that in natural cases of pyometra. In the uninoculated horns, virtually no expression of sex hormone receptors was observed, in contrast to such expression in normal metoestrus. Bacteria-associated ovario-uterine interactions may have been responsible for the hyperplastic (inoculated horn) and inactive (uninoculated horn) uterine changes observed in this experiment. PMID- 12634086 TI - Embolized mesothelial cells within mediastinal lymph nodes of three dogs with idiopathic haemorrhagic pericardial effusion. AB - Embolized mesothelial-like cells were detected within pericardial lymphatics and mediastinal lymph nodes of three golden retrievers with idiopathic haemorrhagic pericardial effusion. Morphological, cytochemical, and immunohistochemical investigations indicated that the embolized cells most likely originated from the pericardial mesothelium. None of the dogs showed evidence of an underlying neoplastic disorder. Such findings have not been reported previously in animals, but so-called "benign mesothelial cell inclusions" have been reported in mediastinal lymph nodes of human patients with pleuritis and pericarditis but no history of neoplasia. The present findings in dogs indicate the need to distinguish between lymphatic emboli arising from reactive mesothelial cells and metastases arising from a mesothelioma. PMID- 12634087 TI - Experimental infection with equine herpesvirus 9 (EHV-9) in cats. AB - The pathogenicity for cats of EHV-9, a new neurotropic equine herpesvirus, was assessed by intranasal inoculation with 10(6) plaque-forming units. Four cats killed 4, 5, 6 or 10 days after inoculation showed neurological signs consisting of hyper-excitability and aggressiveness, followed by tremors, occasional convulsions, and depression. Histologically, the cats showed severe encephalitis characterized by neuronal degeneration and loss, intranuclear inclusions, perivascular cuffing and gliosis in the cerebrum. A positive immunohistochemical reaction for EHV-9 antigen was seen in degenerating neuronal cells. The lesions extended from the olfactory bulb to the rhinencephalon and hippocampus. All cats had rhinitis, with or without intranuclear inclusion bodies in the nasal mucosa, and interstitial pneumonia. These findings indicate that the cat, like certain other species such as the goat, is susceptible to experimental infection with EHV 9, and may be at risk from natural infection. PMID- 12634088 TI - Interactions of Salmonella enterica subsp. enterica serovar Muenchen with intestinal explants of the turtle Trachemys scripta scripta. AB - Salmonella infections in reptiles, in contrast to those in birds and mammals, are limited to the intestinal tract. In this study, interactions of a strain of Salmonella enterica subsp. enterica serovar Muenchen (SEEM) with intestinal explants of the turtle Trachemys scripta scripta were examined by scanning electron microscopy (SEM). Adhesion and invasion in the chelonian intestinal explants at 30 degrees C and 37 degrees C were evaluated quantitatively. For purposes of comparison, the invasive capacity of SEEM in the continuous avian epithelial cell line DIV-1 at 30 degrees C and 37 degrees C was determined. Small numbers of M-like cells were found in the ileum of the turtles. The bacteria adhered mainly to the mucus of the intestinal explants. Only small numbers of salmonellae were associated with epithelial cells. Higher numbers of bacteria adhered at 30 degrees C than at 37 degrees C. Epithelial damage, embedding of bacteria in the epithelial surface and a ruffling-like process were noted only at 37 degrees C. Minimal numbers of salmonellae invaded the explants at 30 degrees C and 37 degrees C. Invasion of DIV-1 cells was greater at 37 degrees C than at 30 degrees C. The study suggested that the intestinal mucous layer provides an important site of colonization for salmonellae in the chelonian host and protects the underlying epithelial cells. PMID- 12634089 TI - Adhesive properties of platelets from different animal species. AB - The use of large animals (e.g., pig and sheep) in human medicine, and the need to develop new therapeutic strategies for domestic animal diseases related to platelet disorders, require better characterization of the physiology of animal platelets. In this study, the ability of platelets from buffaloes, horses, pigs and sheep to adhere to immobilized autologous fibrinogen was compared with that of human platelets. Blood samples were collected from the jugular vein of six healthy subjects of each species and platelet-rich plasma (PRP) was obtained by centrifugation. Platelets, isolated by further centrifugation of PRP, were washed by gel-filtration on Sepharose-2B, counted and added to the wells of 96-well plates pre-coated with autologous fibrinogen. After different times of incubation, non-adherent platelets were removed, and the number of adherent platelets was assessed by measuring endogenous acid phosphatase activity. Horse platelets showed the strongest ability to adhere to autologous immobilized fibrinogen, being 1.7-, 3.1- and 2.3-fold more active than human, buffalo and porcine platelets, respectively. Sheep platelets were unable to adhere to autologous immobilized fibrinogen. Platelet activation by adenosine 5-diphosphate (ADP) increased both human and animal platelet adhesive response. ADP-stimulated sheep platelets were able to adhere to autologous immobilized fibrinogen, albeit to a lesser extent than platelets from the other animal species. The observed interspecies variability in adhesive properties of platelets may reflect structural differences, or differences in the availability of the fibrinogen receptor (glycoprotein IIb/IIIa) on the platelet surface. PMID- 12634090 TI - Cell death and decreased synaptic protein expression in the ventral horn of Holstein-Friesian calves with spinal muscular atrophy. AB - A neuropathological study of Holstein-Friesian calves with spinal muscular atrophy (SMA) demonstrated decreased numbers of motor neurons in the brachial and lumbo-sacral regions of the spinal cord, together with swelling and accumulation of phosphorylated neurofilaments, and neuronophagia in most of the remaining motor neurons. The pyramidal tracts, motor cortex and thalamus were not affected. Synaptophysin immunohistochemistry revealed a marked reduction of punctate terminals but only around swollen neurones, suggesting loss of terminal afferents on motor neurons at advanced stages of the degenerative process. An immunohistochemical study of proteins linked with cell death and cell survival demonstrated reduced expression of Fas, Fas-L, Bcl-2 and Bax in swollen motor neurons. Punctate cytochrome C immunoreactivity, consistent with mitochondrial localization, was detected in the soma of normal motor neurons, but not in swollen motor neurons. Finally, no labelling of motor neurons with antibodies to cleaved (active) caspase-3 (17kD) was detected, suggesting a lack of involvement of the apoptotic pathways in motor neuron death. Taken together, the present findings point to necrosis as a major cause of motor neuron death in the advanced stages of SMA in Holstein-Friesian calves. PMID- 12634091 TI - Apoptosis in calf pneumonia induced by endobronchial inoculation with bovine adenovirus type 3 (BAV-3). AB - Three calves aged 1 week (group 1), three aged 6 weeks (group 2) and three aged 6 weeks (having been pretreated with dexamethasone) (group 3) were infected endobronchially with bovine adenovirus type 3 (BAV-3). All calves had received colostrum. The histopathological, immunohistochemical, ultrastructural and TUNEL features were examined on post-inoculation day (PID) 3, 5 and 7. Viral replication and intranuclear inclusions were frequently observed in groups 1 and 3, but not in group 2. The lesions became progressively severe on PID 5 and 7 in group 1. In group 3, however, the cellular injury caused by BAV-3 was of short duration and the lesions began to resolve at PID 7. Numerous apoptotic cells were seen in the PID 3 calves of all three groups, and in the PID 7 calves of groups 2 and 3; however, the PID 5 and 7 calves of group 1 showed only a few apoptotic cells in the alveolar septa. The results indicated that (1) the durability of BAV 3 infection in the lung was closely related to apoptosis, and (2) the host defence mechanism that induced apoptosis in infected cells was age-related. PMID- 12634092 TI - Characterization of immune cell populations in oral mucosal tissue of healthy adult cats. AB - The aim of this study was to characterize the leucocyte subsets present in the oral mucosa of healthy cats. Immunohistochemical labelling and computer-assisted morphometric analysis was used to identify expression of MHC class II, CD3, CD79a, IgG, IgM, IgA, and leucocyte antigen L1 (L1) by cells in sections from 19 cats, and expression of CD4 and CD8 by cells in sections from 17 cats. Mast cells were detected by toluidine blue staining. In the epithelial compartment, CD3(+) intraepithelial lymphocytes were detected, and CD8(+) cells were more common than CD4(+) cells. MHC class II labelling revealed intraepithelial and subepithelial cells with a characteristic dendritic morphology. In some sections these dendritic cells were closely associated with subepithelial clusters of CD3(+) T cells containing both CD4(+) and CD8(+) cells. In the lamina propria and submucosal compartments, the cells most commonly identified were mast cells. CD3(+) T-lymphocytes were also observed, and CD4(+) and CD8(+) cells were detected in similar numbers. L1(+) and CD79(+) cells were detected least frequently. The few plasma cells present were generally found to be either IgG(+) or IgA(+). Within the stroma surrounding the salivary glands, CD79a(+) and IgA(+) cells predominated. Slight epithelial labelling for L1 was seen in some sections. The normal feline oral mucosa clearly contains a range of immune cell populations. PMID- 12634093 TI - Early pathogenesis and inflammatory response in experimental bovine mastitis due to Streptococcus uberis. AB - A generally similar clinical response was observed in six lactating Holstein Friesian cows after intramammary inoculation with approximately 10(7) colony forming units of Streptococcus uberis. Increased concentrations of serum amyloid A (SAA) were measured in both milk and serum taken 6 and 11h after inoculation, respectively. In contrast, increased concentrations of haptoglobin were detected after 10h of infection, in milk only. In the blood, tumour necrosis factor-alpha (TFN-alpha) was detected (0.503 ng/ml) in only one animal, at the time of euthanasia (10h after infection). Interferon-gamma (IFN-gamma), like haptoglobin, was not detected in blood. Parallel to the development of inflammation and influx of inflammatory cells into the udder tissue, a marked decrease in the number of monocytes and neutrophils in blood was observed. Bacteria were found both intracellularly (macrophages and neutrophils) and within the lumen of ducts and alveoli. Lesions developed progressively in an ascending manner and became widespread throughout the mammary gland in less than 8h. The parallel development of inflammation and increased concentrations of SAA and haptoglobin in milk points to these acute phase proteins as potential diagnostic markers for the early detection of S. uberis -associated mastitis. PMID- 12634094 TI - Collateral circulation in experimental coarctation of the aorta in minipigs: a possible association with hypertrophied vasa vasorum. AB - Experimental coarctation of the thoracic aorta was produced in 13 adult Yucatan minipigs by placing a gradually expanding "C-shaped" occluder around the aorta. The aortic constriction was standardized by measuring the blood pressure above the coarctation, with a mean arterial pressure of 165-170mmHg as the target. The pigs were humanely killed after 8 weeks of hypertension. No deaths were associated with constriction of the aorta, and neither rear limb weakness nor neurological dysfunction was noted. The lack of clinical signs in these animals suggested the development of an extensive collateral circulation. Collateral vessels, which were verified angiographically, included intercostal arteries, muscular intercostal arteries, and the internal thoracic artery. In two of the animals in which the thoracic aorta was completely occluded, revascularization also occurred via small arterial channels. Histologically, there was a marked increase in the number of vessels as well as significant hyperplasia and hypertrophy of smooth muscle of adventitial vasa vasorum. These findings support the hypothesis that collateralization of occluded thoracic aortas may be aided by hypertrophy of the vasa vasorum. To our knowledge, these vessels have not previously been shown convincingly by histopathological methods to revascularize occluded large arteries such as the aorta. PMID- 12634095 TI - Detection of PrPSc in lymphoid tissues of lambs experimentally exposed to the scrapie agent. AB - Histoblotting and immunohistochemistry were used to detect disease-associated prion protein (PrP(Sc)) in lymphoid tissues of lambs of known PrP genotype infected with the scrapie agent by stomach tube at the age of 2 months. The ileal and jejunal Peyer's patches and retropharyngeal and distal jejunal lymph nodes were studied 1 week, 5 weeks, 5 months and 11 months after inoculation. Other lymphoid tissues examined included superficial cervical lymph node, tonsil and spleen. PrP(Sc) was not detected in any tissue of any lamb at 1 week post inoculation. At 5 weeks, PrP(Sc) was detected in tissues of lambs of susceptible PrP genotypes (AV(136)QQ(171) and VV(136)QQ(171)), but not lambs of other PrP genotypes (AA(136)QQ(171), AA(136)QR(171) and AV(136)QR(171)). PrP(Sc) was present in the germinal centres of tonsils, distal jejunal and retropharyngeal lymph nodes, and spleen. In the nodules of ileal and jejunal Peyer's patches, only occasional solitary cells showed the presence of PrP(Sc). At 5 months post inoculation, increased accumulations of PrP(Sc) were detected in ileal and jejunal Peyer's patches, as well as in the retropharyngeal and distal jejunal lymph nodes of a single lamb inoculated with the agent from a sheep of the same susceptible PrP genotype. Eleven months after exposure to the scrapie agent, PrP(Sc) was detected in all lymphoid tissues examined from sheep of susceptible PrP genotypes. These studies show that PrP(Sc) was detectable in lymphoid tissues 5 weeks after exposure to the scrapie agent by stomach tube in lambs as young as 3 months of age and indicate that the PrP genotype is a significant factor for the rapid uptake and spread of the agent through lymphoid tissues. PMID- 12634096 TI - Distribution of ehrlichiae in tissues as determined by in-situ hybridization. AB - Specific identification of ehrlichiae in the tissues and determination of their distribution is difficult. In this study, an in-situ hybridization method was developed to detect ehrlichial 16S rRNA in tissue specimens from mice experimentally infected with the HF strain. This strain is closely related to Ehrlichia chaffeensis, the agent of human monocytic ehrlichiosis. HF strain specific 16S rRNA was detected in endothelial cells and monocyte-macrophages in the liver, lungs, bone marrow, spleen, lymph nodes, and large and small intestinal tissues. The results suggest that the in-situ hybridization method with a digoxigenin-labelled RNA probe specific to ehrlichial 16S rRNA will be useful for post-mortem diagnosis and for the histopathological investigation of ehrlichial infection. PMID- 12634097 TI - The pathogenicity of diminazene aceturate-resistant Trypanosoma brucei in rats after treatment with the drug. AB - Four groups (A, B, C and D) of 10 rats were used to determine the effect of comparatively high doses of diminazene aceturate on diminazene aceturate resistant Trypanosoma brucei and the pathogenic effect of relapse infection. Group A rats were uninfected (controls) while group B, C and D rats were inoculated intraperitoneally with 0.5 x 10 (6) diminazene aceturate-resistant T. brucei and treated with diminazene aceturate at 14.0, 17.5 and 21.0mg/kg body weight, respectively, on day 14 post-infection (PI) as a single intraperitoneal injection. Prepatent periods and also levels of parasitaemia were comparable in groups B, C and D. Packed cell volume (PCV) decreased in the infected groups by day 14 PI and returned to pre-infection values by day 63 post-treatment (PT). Anaemia was comparable in groups B, C and D. Relapse parasitaemia occurred in six rats in group B on day 70 PT and in five rats in each of groups C and D on day 77 PT. The PCV of the rats with relapse infection decreased progressively up to day 105 PT, when the experiment was terminated, whereas the PCV of rats without relapse did not. The levels of anaemia and parasitaemia on day 14 post-relapse were significantly higher (P<0.05) than the levels obtained on day 14 PI in the same animals. Thus, comparatively high doses of diminazene aceturate failed to cure drug-resistant T. brucei infection in 50-60% of infected rats and relapse infections were more severe than the primary infections before treatment. PMID- 12634098 TI - Hepatocellular endoplasmic reticulum storage disease in an African elephant (Loxodonta africana). AB - Large intracytoplasmic inclusions were observed in hepatocytes of a 7-year-old African elephant (Loxodonta africana). The inclusions were oval to polyhedral with either a homogeneous glassy or a granular appearance. They were positive for the periodic acid-Schiff (PAS) reaction. Electron microscopical examination revealed that the inclusions consisted of granular material with moderate electron-density and were membrane-bounded. The findings suggested that the inclusions were derived from endoplasmic reticulum. The light and electron microscopical features were similar to those of endoplasmic reticulum storage disease of the liver in man. Such inclusions have not previously been reported in animals. PMID- 12634099 TI - Cerebral primitive neuroectodermal tumour in a heifer. AB - A primitive neuroectodermal tumour (PNET) replacing the thalamus was discovered in an 18-month-old Prim'Holstein heifer. Microscopical examination of the tumour showed large sheets of densely packed cells with occasional Homer-Wright and perivascular rosettes. Neoplastic cells were small with ill-defined borders, scant cytoplasm and ovoid, irregularly shaped nuclei. Immunolabelling was positive for vimentin and neuron-specific enolase, in agreement with previous reports of PNETs in human beings and animals. This appears to be the first report of cerebral PNET in cattle. PMID- 12634100 TI - Glomus tumour in the digit of a dog. AB - The gross and microscopical features of a glomus tumour in the digit of a 9-year old dog are described. The tumour consisted of a red nodule near the nail of the third digit of the right forelimb and appeared painful. The tumour cells, which had round to oval hyperchromatic nuclei and scant cytoplasm, were arranged in sheets around blood vessels, or in nests or duct-like structures. This pattern has not been described previously in canine glomus tumours. Mitotic figures were seen only occasionally. Tumour cells were strongly immunolabelled for vimentin and some expressed smooth-muscle actin and desmin. They were negative for cytokeratins, neuron-specific enolase and CD34. Silver impregnation (reticulin method) stained the reticulum around blood vessels, nests of tumour cells and duct-like structures, and a delicate reticulum was seen around each tumour cell. The morphological, immunohistochemical and histochemical patterns helped in the diagnosis of this glomus tumour. PMID- 12634101 TI - Bovine papillomavirus type 4 in oesophageal papillomas of cattle from the south of Italy. AB - Oesophageal papillomas are known to occur in cattle infected with bovine papillomavirus type 4 (BPV-4), and BPV-4 papillomas may undergo malignant progression in cattle that feed on bracken fern. In the south of Italy, where bracken fern is common, examination of 1133 slaughterhouse cattle aged 4-12 years revealed oesophageal lesions (single or multiple peduncuolated proliferations, or mucosal thickening) in 147 (13%). These two types of lesion were consistent with exophytic and inverted papilloma, respectively. BPV-4 was detected by polymerase chain reaction (PCR) analysis in >60% of the samples in which oesophageal papilloma was diagnosed histopathologically. Nucleotide sequencing of the PCR amplicons confirmed the presence of BPV-4 in the papillomas. This is the first report of such infections in a European country other than Britain. PMID- 12634102 TI - Emboli in bulls killed in Spanish traditional bullfighting. AB - The finding of brain tissue fragments in blood and lungs of cattle stunned in slaughterhouses has raised concerns about food safety in the context of the bovine spongiform encephalopathy epidemic. In the present study, the possible occurrence of brain tissue emboli in animals killed in traditional Spanish bullfighting was investigated. Thorough histological analysis of multiple possible target organs was carried out in 434 bulls. No evidence of brain tissue embolism was obtained, but emboli from diverse sources were detected in pulmonary and hepatic tissue of a significant number of animals. These emboli seem to have been caused by the use of a long sword, which extensively disrupts intra-thoracic and intra-abdominal organs and vascular structures. PMID- 12634103 TI - Ameloblastic carcinoma in a horse. AB - The clinical, gross morphological, histological and immunohistochemical characteristics of an ameloblastic carcinoma in a 30-year-old Quarter Horse mare are reported. This tumour was fast growing, locally invasive and destructive. Histologically, it showed an infiltrative pattern of large islands, broad sheets and, at the periphery, small cords of moderately pleomorphic round, oval to spindle-shaped cells. Immunohistochemical evaluation revealed positive labelling for vimentin, cytokeratin 5/6 and cytokeratin 14. In the oral cavity of human beings, this immunolabelling pattern is unique for the embryonal enamel organ and tumours of ameloblastomatous epithelial origin, which strongly supports the diagnosis of equine ameloblastic carcinoma. PMID- 12634104 TI - Oxidative stress-induced mitochondrial DNA damage in human retinal pigment epithelial cells: a possible mechanism for RPE aging and age-related macular degeneration. AB - Oxidative stress is believed to contribute to the pathogenesis of many diseases, including age-related macular degeneration (AMD). Although the vision loss of AMD results from photoreceptor damage in the central retina, the initial pathogenesis involves degeneration of RPE cells. Evidence from a variety of studies suggests that RPE cells are susceptible to oxidative damage. Mitochondrial DNA (mtDNA) is particularly prone to oxidative damage compared to nuclear DNA (nDNA). Using the quantitative PCR assay, a powerful tool to measure oxidative DNA damage and repair, we have shown that human RPE cells treated with H(2)O(2) or rod outer segments resulted in preferential damage to mtDNA, but not nDNA; and damaged mtDNA is not efficiently repaired, leading to compromised mitochondrial redox function as indicated by the MTT assay. Thus, the susceptibility of mtDNA to oxidative damage in human RPE cells, together with the age-related decrease of cellular anti-oxidant system, provides the rationale for a mitochondria-based model of AMD. PMID- 12634105 TI - Ultrastructural characterization and Fourier analysis of fiber cell cytoplasm in the hyperbaric oxygen treated guinea pig lens opacification model. AB - The structural characteristics of differentiated fiber cells in control and hyperbaric oxygen (HBO)-treated guinea pig lenses were examined by transmission electron microscopy (TEM). Emphasis was placed on cell damage, membrane integrity, and cytoplasmic texture. Given the faint gross opacities observed in HBO-treated lenses in previous studies, it was hypothesized that subtle but significant morphological differences due to oxidative damage exist between control and treated animals. Experimental animals received either 70 or 85 treatments with HBO (2.5 atm of 100% O(2) for 2.5 hr, 3 times per week for 5-7 months). All specimens were obtained within 24 hr of death. Freshly cut Vibratome lens sections were fixed and processed for low and high-magnification thin section TEM analysis. Cytoplasmic texture was analyzed using Fourier and autocorrelation image processing techniques. Low-magnification analysis revealed relatively insignificant differences in general appearance between the fiber cells of the inner fetal and embryonic nuclei in control and HBO-treated guinea pigs. Both groups demonstrated cells of similar morphology with equivalent membrane complexity and homogeneous cytoplasmic texture. Evidence of any major cellular damage or extracellular space debris was not obvious. High-magnification analysis of the cytoplasm of the treated lenses exhibited a mild, yet detectable increase in texture compared with controls and was confirmed by Fourier analysis. Cytoplasmic texture increased in complexity with additional treatments. The absence of major cellular damage in the lenses of HBO-treated animals suggests a less conspicuous source of light scattering. The small changes in cytoplasmic organization observed between treated and control animals may entirely account for the increase in nuclear light scattering observed by slit lamp. The results obtained with this guinea pig/HBO model parallel many of the morphological data associated with human nuclear cataracts. The high-angle scattering observed in the lens of the HBO-treated guinea pig may represent the type of cytoplasmic reorganization that occurs with mild oxidation, effectively making it a valuable model for human lens aging. PMID- 12634106 TI - Minocycline effect on meibomian gland lipids in meibomianitis patients. AB - The objective of this research was to determine the effect of oral minocycline on the meibomian gland nonpolar and free fatty acid lipids of chronic blepharitis patients. Patients--seborrheic blepharitis (SBBL), acne rosacea (AR) without ocular involvement, and acne rosacea with meibomianitis (AR-MKC). Minocycline treatment--50mg orally for 2 weeks followed by 100mg to the end of 3 months; this was followed by 3 more months with no treatment. Meibomian gland secretions (meibum) were collected before treatment, at the end of the 3 months on treatment, and 3 months after stopping treatment. Lipids were separated and analyzed for wax and sterol esters, triglycerides, diglycerides, free cholesterol and free fatty acids. Data were analyzed statistically by ANOVA. Minocycline treatment resulted in decreased diglycerides and free fatty acids in the group AR MKC, which continued into the second 3 months (off treatment) and was significant. Cholesterol decreased, but triglycerides initially decreased with treatment and then increased when treatment in the group was discontinued (second 3 months); these results, however, were not significant. Thus, minocycline has its greatest effect on lipid types, which result from degradation (lipase) reactions, suggesting a lipase inhibition effect and/or direct effect on ocular flora. This minocycline effect continues even after treatment is discontinued, suggesting a more lasting effect on ocular microflora. Minocycline may be most effective when the treatment period is longer than 3 months. These results give insight into disease mechanisms associated with chronic blepharitis. PMID- 12634107 TI - Expression and activation of STAT proteins during mouse retina development. AB - Cytokines and growth factors play important roles in mammalian ocular development and maintenance. Recent studies have indicated that some of these ligands can activate signal transducer and activator of transcription factors (STATs) and modulate gene transcription. The purpose of this study was to investigate the expression and activation of STAT proteins in the developing mouse retina. Anti STAT and anti-phosphorylated STAT antibodies were used to detect the expression and activation of STATs in embryonic and postnatal neuronal retina, ciliary margin, and retinal pigment epithelium (RPE). In situ hybridization and Western blot were also employed. In embryonic stages, all STAT proteins were expressed in the neuronal retina in distinct cell populations at different embryonic stages. For example, Stat3 expression and activation gradually increased in the inner neuroblast layer and ciliary margin during development. In adult retina, Stat3 was detected in the inner nuclear layer and ganglion cells layers. Stat1 was strongly expressed in both outer and inner plexiform layers. Stat5a was clearly expressed in the outer/inner nuclear layer, the ganglion cell layer, and the inner plexiform layer. Strong expression of Stat3, Stat5a, and Stat6 was observed in the RPE. Activated Stat3 and Stat5a were found in the neural retina and the RPE. Distinct STAT proteins were present in different cell populations in neuronal retina and RPE suggesting multiple functions of STATs in mammalian eye development. Studies of STAT signal pathways in the eye may contribute to the understanding of molecular mechanisms in control of ocular development and pathogenesis. PMID- 12634108 TI - Protein expression patterns for ubiquitous and tissue specific calpains in the developing mouse lens. AB - Calcium activated proteases (calpains) have been implicated in the processing of lens crystallins during lens maturation and cataract formation. Ubiquitous type calpain 2 and calpain 10 and lens specific Lp82 and Lp85 protein distribution were determined using immunohistochemistry and immunoblotting in embryonic and post-natal mouse eyes. Calpain 2 was first expressed late in embryonic development and localized to the lens epithelium and transition zone. Lp82 was expressed at E9.5 in the lens placode, head ectoderm, and throughout the fiber cells during embryonic lens maturation. Lp82 co-localized at sites of crystallin modification in the juvenile lens. In the adult lens, Lp82 protein was maintained in cortical fibers but could not be detected in the lens nucleus. Lp85, the slightly larger splice variant of Lp82, was first observed at E9.5 and throughout early embryonic lens development. Abundant localization of this enzyme was observed in the cell nuclei of lens epithelium, elongating fibers, and undifferentiated mesoderm. Robust peri-nuclear localization of calpain 10 was observed in the head ectoderm, lens placode, and optic vesicle during early eye induction. Further, calpain 10 protein was maintained in the lens epithelium of pre- and post-natal lens. These data support the hypothesis that Lp82 in rodent lens has an important role in crystallin proteolysis during normal lens maturation. In contrast, calpain 2, Lp85, and calpain 10 may have roles in cell signaling pathways. PMID- 12634109 TI - Effect of MCI-9042, a 5-HT2 receptor antagonist, on retinal ganglion cell death and retinal ischemia. AB - The neuroprotective effect of MCI-9042 (Mitsubishi Pharma Corporation) was investigated on glutamate-induced retinal ganglion cell (RGC) death in vitro and on rat retinal ischemia in vivo. RGCs were purified from retinal cells isolated from 6-day-old Wistar rats and cultured in serum-free media. After application of 25 microM glutamate, the viability of RGCs treated with or without several serotonin 2 (5-HT(2)) receptor antagonists: MCI-9042, M-1 (a major metabolite of MCI-9042), ketanserin, and LY-53857; was evaluated by calcein-acetoxymethyl ester staining. Retinal ischemia was induced by intraocular pressure (IOP) elevation (130 mmHg, 50 min). Rats were intraperitoneally injected with MCI-9042 at a dose of 3, 30 mg/kg or base at 30 min before and just after ischemia-reperfusion. Retinal damages were evaluated by histology, morphometric analysis and electroretinograms (ERGs) recordings at 7 days after ischemia-reperfusion. 25 microM glutamate decreased the number of viable RGCs to about 60 to 65% of untreated RGCs. MCI-9042, M-1, ketanserin, and LY-53857 significantly reduced glutamate-induced RGC death at concentrations of more than 100 nM, 1 nM, 1 microM and 100 nM, respectively. Ischemia-reperfusion caused thinning of the thickness between the inner plexiform layer and the outer plexiform layer and attenuation of a-and b-waves in ERG recordings. The intraperitoneal injection of MCI-9042 significantly reduced morphological and functional damages in retinal ischemia. Our data demonstrate that 5-HT(2) receptor antagonists including MCI-9042 and M-1 have the neuroprotective effects in cultured RGCs and that MCI-9042 protects against ischemic retinal diseases. PMID- 12634110 TI - L-NAME protects against acute light damage in albino rats, but not against retinal degeneration in P23H and S334ter transgenic rats. AB - Two previous studies have shown that N(G)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of neuronal nitric oxide synthase, protects retinas of albino rats and mice from damaging levels of light. The aims of the present study were two fold: (1) to confirm the protective effect of L-NAME on wild type albino rats and (2) to determine if L-NAME protects the retinas of transgenic rats with P23H and S334ter rhodopsin mutations. In the first study, albino rats born and raised in 5 10 lux cyclic light were injected intraperitoneally with either L-NAME or its inactive isomer D-NAME 30 min before being placed in bright light (2700 lux) for 24hr. Electroretinograms (ERGs) were recorded before light treatment and 2 days after cessation of exposure, and eyes were enucleated for morphologic evaluation. L-NAME, but not D-NAME provided structural protection of photoreceptor cells from light damage. The functional rescue was not statistically significant between the drug treated groups. In the second study, albino WT, P23H transgenic, and S334ter transgenic rats were born and raised in 400 lux cyclic light. Three week old animals received daily intraperitoneal injections of L-NAME or D-NAME for 4 weeks, and the same drugs were added to their drinking water. At 7 weeks of age, the ERG sensitivity curves and the outer nuclear layer thickness of both transgenic groups were significantly reduced compared to WT controls. However, administration of L-NAME did not protect against retinal degeneration caused by the rhodopsin mutation in either strain of transgenic (P23H and S334ter) rats. Thus, although photoreceptor cell death in light damage and inherited retinal degenerations share a common apoptotic mechanism, there must be significant 'up stream' differences that allow selective neuroprotection by L-NAME. PMID- 12634111 TI - Activated microglia in human retinitis pigmentosa, late-onset retinal degeneration, and age-related macular degeneration. AB - Many gaps exist in our knowledge of human retinal microglia in health and disease. We address the hypothesis that primary death of rod photoreceptors leads to activation of resident microglia in human retinas with retinitis pigmentosa (RP), late-onset retinal degeneration (L-ORD), or age-related macular degeneration (AMD). Regions of ongoing photoreceptor cell death were studied by immunocytochemistry with microglia- and other retinal cell-specific markers. In normal human retinas, quiescent microglia were small, stellate cells associated with inner retinal blood vessels. In retinas with RP, L-ORD, or AMD, numerous activated microglia were present in the outer nuclear layer in regions of ongoing rod cell death. These microglia were enlarged, amoeboid cells that contained rhodopsin-positive cytoplasmic inclusions. We conclude that activated microglia migrate to the outer nuclear layer and remove rod cell debris. In other central nervous system diseases such as stroke, activated microglia phagocytose debris from the primary injury and also secrete molecules that kill nearby normal neurons. By analogy with these diseases, we suggest that microglia activated by primary rod cell death may kill adjacent photoreceptors. Activated microglia may be a missing link in understanding why initial rod cell death in the human diseases RP, L-ORD, and AMD leads to death of the cones that are critical for high acuity daytime vision. PMID- 12634112 TI - Ultrastructural analysis of hydraulic and abrasive retinal pigment epithelial cell debridements. AB - Differential changes in Bruch's membrane, choriocapillaris, retinal pigment epithelium, retina, and tapetum after hydraulic or abrasive debridement of the retinal pigment epithelium in the cat area centralis were documented by fluorescein angiography, histology, and transmission electron microscopy at 1 hour, 1-day, 3-day, 1-week, or 4-week time points. Abrasive debridement is associated with abnormal fluorescein angiography and incomplete ingrowth of retinal pigment epithelial cells. Transmission electron microscopy shows that abrasive debridement inflicts more long-lasting ultrastructural damage to Bruch's membrane, the choriocapillaris, tapetum, and retina than does hydraulic debridement. Because the retinal pigment epithelium can resurface abrasively debrided Bruch's membrane that is disorganized, split, reduplicated, or missing, we cannot correlate the ultrastructural appearance of Bruch's membrane with the likelihood of complete resurfacing of the debrided area. Primary choriocapillary or retinal damage in abrasive debridements may contribute to the poor outcome. Regions of retinal degeneration with no underlying retinal pigment epithelial cell monolayer were significantly larger in abrasive debridements at the 4-week than at the 1-week time point. Reduced resurfacing at the later time point suggests that not all cells resurfacing abrasively debrided areas survived over the longer term. This finding may mean that retinal pigment epithelial cells are not able to resurface completely and permanently areas showing geographic atrophy of the choriocapillaris. PMID- 12634113 TI - Cotransport of H+, lactate, and H2O in porcine retinal pigment epithelial cells. AB - The retinal pigment epithelium (RPE) of the eye transports water and lactate ions in the direction from retina to choroid. The water transport is important in maintenance of retinal adhesion and the transport of lactate ions serves to regulate the lactate levels and pH of the subretinal space. This study investigates by means of a non-invasive technique the mechanism of coupling between transport of H(+), lactate ion, and water in the monocarboxylate transporter (MCT1) located in the apical (retinal) membrane of a mammalian RPE. Primary cultures of porcine RPE cells were grown to confluence and placed in a perfusion chamber in which the solution facing the retinal membrane could be changed rapidly. Two types of experiments were performed: Changes in cell water volume were measured by self-quenching of the fluorescent dye Calcein, and changes in intracellular pH were measured ratiometrically using the fluorescent dye BCECF. In lactate-free solutions, mannitol addition to the retinal bath caused intracellular acidification and cell shrinkage, given by a single osmotic water permeability of 1.2+/-0.1 x 10(-4)cmsec(-1) (osmoll(-1))(-1). In solutions containing 50 mmoll(-1) lactate, however, the mannitol-induced cell shrinkage was faster and the cells alkalinized. These effects were not linear functions of the magnitude of the imposed osmotic gradients: Both volume effects and changes in intracellular pH showed apparent saturation with increasing gradients. Abrupt isosmotic replacement of Cl(-) with lactate in the concentration range from 3 to 50 mmoll(-1) caused an immediate cell swelling as well as an immediate intracellular acidification; both effects showed apparent saturation with increasing lactate concentration. The K(m) values were: 11+/-2 mmoll(-1) for the water fluxes and 13+/-4 mmoll(-1) for the H(+) and lactate fluxes. The data suggest that H(2)O is cotransported along with H(+) and lactate ions in MCT1 localized to the retinal membrane. The study emphasizes the importance of this cotransporter in the maintenance of water homeostasis and pH in the subretinal space of a mammalian tissue and supports our previous study performed by an invasive technique in an amphibian tissue. PMID- 12634114 TI - The beta-adrenoceptor antagonists metipranolol and timolol are retinal neuroprotectants: comparison with betaxolol. AB - beta-adrenoceptor antagonists are used clinically to reduce elevated intraocular pressure in glaucoma which is characterised by a loss of retinal ganglion cells. Previous studies have shown that the beta(1)-selective adrenoceptor antagonist, betaxolol, is additionally able to protect retinal neurones in vitro and ganglion cells in vivo from the detrimental effects of either ischemia-reperfusion or from excitotoxicity, after topical application. The neuroprotective effect of betaxolol is thought not to be elicited through an interaction with beta adrenoceptors, but by its ability to reduce influx of sodium and calcium through voltage-sensitive calcium and sodium channels. In the present study it is shown that the non-selective beta-adrenoceptor antagonists, metipranolol and timolol behave like betaxolol. When topically applied they all attenuate the detrimental effect of ischemia-reperfusion. Protection of the retina was determined by evaluating changes in the electroretinogram and by assessing the loss of mRNA for Thy-1, which is expressed in retinal ganglion cells. In addition, studies conducted on neurones in mixed retinal cultures demonstrated that metipranolol, betaxolol and timolol were all able to partially counteract anoxia-induced cell loss and viability reduction. The influence of timolol was, however, not significant. Within the confines of these investigations, an order of neuroprotective efficacy was delineated for the three beta-adrenoceptor antagonists: betaxolol>metipranolol>timolol. The ability of the beta-adrenoceptor antagonists to attenuate ligand-induced stimulation of calcium and sodium entry into neuronal preparations showed a similar order of effectiveness. In conclusion, the ability to confer neuroprotection to retinal neurones is a common feature of three ophthalmic beta-adrenoceptor antagonists (betaxolol, metipranolol and timolol). A comparison of the effectiveness of the individual compounds in protecting retinal cells in vivo was not possible in these studies. However, in vitro studies show that the capacity of the individual beta adrenoceptor antagonists to act as neuroprotectants appears to relate to their capacity to attenuate neuronal calcium and sodium influx. PMID- 12634115 TI - Chemical mixtures research: significance and future perspectives. PMID- 12634116 TI - International issues on human health effects of exposure to chemical mixtures. AB - In this article, we highlight new developments and recent studies concerning adverse human health effects related to chemical mixtures. One group of activities comprises the development of a new computer program for analyzing mixture studies and a mathematical model as a basis for combination rules that predict the toxicity of mixtures. Other new activities in the area of experimental studies are the application of gene expression technologies in mixture research, and pattern recognition as a tool in safety evaluation of complex mixtures. A "bottom-up" approach for chemosensory detection of mixtures has recently been presented. Other topics include a method for the safety evaluation of natural flavoring complexes, and an evaluation of the possible health effects of the simultaneous intake of food additives. Examples of issues related to mixtures of airborne chemicals are potential interaction of fine particles and gaseous pollutants in ambient air, nasal cancer associated with inhaled chemical mixtures, and the recommendation of a limit value for volatile organic compounds. Topics of a more strategic nature include studies concerning the public health effects of large airports, and the development of criteria for a harmonized classification of chemical mixtures. This overview illustrates that strategies to tackle the safety evaluation of combined exposures and complex mixtures as well as models facilitating the interpretation of findings in the context of risk assessment of mixtures have become increasingly important. It is true that exposure of humans to chemical mixtures is the rule rather than the exception, and therefore health risk assessments should focus on mixtures and not on single chemicals. It is also true, however, that humans have learned to cope with exposure to huge numbers of chemicals simultaneously (food, water, air, soil, and consumer products). Therefore, in view of limited resources for toxicological research, the focus in toxicology should be on priority mixtures- priority being determined by (estimated) health risk (= toxicity and exposure). PMID- 12634118 TI - The midgut epithelium of aquatic arthropods: a critical target organ in environmental toxicology. AB - The midgut epithelium of aquatic arthropods is emerging as an important and toxicologically relevant organ system for monitoring environmental pollution. The peritrophic matrix of aquatic arthropods, which is secreted by the midgut epithelium cells, is perturbed by copper or cadmium. Molecular biological studies have identified and characterized two midgut genes induced by heavy metals in the midgut epithelium. Many other metal-responsive genes (MRGs) await characterization. One of the MRGs codes for an intestinal mucin, which is critical for protecting the midgut from toxins and pathogens. Another codes for a tubulin gene, which is critical for structure and function of the midgut epithelial cells. Perturbation of expression of either gene could condition aquatic arthropod survivorship. Induction of these MRGs is a more sensitive and rapid indicator of heavy-metal pollution than biological assays. Characterization of genes induced by pollutants could provide mechanistic understanding of fundamental cellular responses to pollutants and insight into determinants of aquatic arthropod population genetic structure and survivorship in nature. PMID- 12634119 TI - Deviation from additivity in mixture toxicity: relevance of nonlinear dose response relationships and cell line differences in genotoxicity assays with combinations of chemical mutagens and gamma-radiation. AB - Sublinear dose-response relationships are often seen in toxicity testing, particularly with bioassays for carcinogenicity. This is the result of a superimposition of various effects that modulate and contribute to the process of cancer formation. Examples are saturation of detoxification pathways or DNA repair with increasing dose, or regenerative hyperplasia and indirect DNA damage as a consequence of high-dose cytotoxicity and cell death. The response to a combination treatment can appear to be supra-additive, although it is in fact dose-additive along a sublinear dose-response curve for the single agents. Because environmental exposure of humans is usually in a low-dose range and deviation from linearity is less likely at the low-dose end, combination effects should be tested at the lowest observable effect levels (LOEL) of the components. This principle has been applied to combinations of genotoxic agents in various cellular models. For statistical analysis, all experiments were analyzed for deviation from additivity with an n-factor analysis of variance with an interaction term, n being the number of components tested in combination. Benzo[a]pyrene, benz[a]anthracene, and dibenz[a,c]anthracene were tested at the LOEL, separately and in combination, for the induction of revertants in the Ames test, using Salmonella typhimurium TA100 and rat liver S9 fraction. Combined treatment produced no deviation from additivity. The induction of micronuclei in vitro was investigated with ionizing radiation from a 137Cs source and ethyl methanesulfonate. Mouse lymphoma L5178Y cells revealed a significant 40% supra additive combination effect in an experiment based on three independent replicates for controls and single and combination treatments. On the other hand, two human lymphoblastoid cell lines (TK6 and WTK1) as well as a pilot study with human primary fibroblasts from fetal lung did not show deviation from additivity. Data derived from one cell line should therefore not be generalized. Regarding the testing of mixtures for deviation from additive toxicity, the suggested experimental protocol is easily followed by toxicologists. PMID- 12634117 TI - Overview of human health and chemical mixtures: problems facing developing countries. AB - In developing countries, chemical mixtures within the vicinity of small-scale enterprises, smelters, mines, agricultural areas, toxic waste disposal sites, etc., often present a health hazard to the populations within those vicinities. Therefore, in these countries, there is a need to study the toxicological effects of mixtures of metals, pesticides, and organic compounds. However, the study of mixtures containing substances such as DDT (dichlorodiphenyltrichloroethane, an insecticide banned in developed nations), and mixtures containing contaminants such as fluoride (of concern only in developing countries) merit special attention. Although the studies may have to take into account simultaneous exposures to metals and organic compounds, there is also a need to consider the interaction between chemicals and other specific factors such as nutritional conditions, alcoholism, smoking, infectious diseases, and ethnicity. PMID- 12634120 TI - Application of genomics to toxicology research. AB - Traditional models of toxicity have relied on dissecting chemical action into pharmacokinetic and pharmacodynamic processes. However, the integration of genomic information with toxicology will enhance our basic understanding of these processes and significantly change the way we apply toxicological information to risk assessment and regulatory problems. In this article, we summarize the application of gene expression information and polymorphism discovery to four areas in toxicology: toxicity testing, cross-species extrapolation, understanding mechanism of action, and susceptibility. PMID- 12634123 TI - The role of cell bioaugmentation and gene bioaugmentation in the remediation of co-contaminated soils. AB - Soils co-contaminated with metals and organics present special problems for remediation. Metal contamination can delay or inhibit microbial degradation of organic pollutants such that for effective in situ biodegradation, bioaugmentation is necessary. We monitored the degradation of 2,4 dichlorophenoxyacetic acid (2,4-D) or 3-chlorobenzoate (3-CB) in two different soils with and without cadmium (Cd) contamination. Additionally, we evaluated the ability of bioaugmentation to enhance organic degradation in these co contaminated soils. Finally, we determined whether enhanced degradation was due to survival of the introduced organism (cell bioaugmentation) or plasmid transfer to indigenous microbial populations (gene bioaugmentation). In Brazito soil, dual inoculation with a Cd-resistant bacterium plus a known 2,4-D-degrading bacterium, Ralstonia eutropha JMP134, enhanced 2,4-D degradation. Escherichia coli D11, which lacks chromosomal genes necessary for complete 2,4-D mineralization, was used for gene bioaugmentation in Madera soil. Significant gene transfer of the plasmid to the indigenous populations was observed, and the rate of 2,4-D degradation was enhanced relative to that of controls. Cell bioaugmentation was further demonstrated when (Comamonas testosteroni was used to enhance biodegradation of 3-CB in Madera soil. In this case no transfer of plasmid pBRC60 to indigenous soil recipients was observed. For the Madera soil, nonbioaugmented samples ultimately showed complete 2,4-D degradation. In contrast, nonbioaugmented Brazito soils showed incomplete 2,4-D degradation. These studies are unique in showing that both cell bioaugmentation and gene bioaugmentation can be effective in enhancing organic degradation in co-contaminated soils. Ultimately, the bioaugmentation strategy may depend on the degree of contamination and the time frame available for remediation. PMID- 12634121 TI - Problems for risk assessment of endocrine-active estrogenic compounds. AB - Estrogenic industrial compounds such as bisphenol A (BPA) and nonylphenol typically bind estrogen receptor (ER) alpha and ERBeta and induce transactivation of estrogen-responsive genes/reporter genes, but their potencies are usually greater than or equal to 1,000-fold lower than observed for 17Beta-estradiol. Risk assessment of estrogenic compounds on the basis of their potencies in simple reporter gene or binding assays may be inappropriate. For example, selective ER modulators (SERMs) represent another class of synthetic estrogens being developed for treatment of hormone-dependent problems. SERMs differentially activate wild type ERalpha and variant forms expressing activation function 1 (ER-AF1) and AF2 (ER-AF2) in human HepG2 hepatoma cells transfected with an estrogen-responsive complement C3 promoter-luciferase construct, and these in vitro differences reflect their unique in vivo biologies. The HepG2 cell assay has also been used in our laboratories to investigate the estrogenic activities of the following structurally diverse synthetic and phytoestrogens: 4 -hydroxytamoxifen; BPA; 2 ,4 ,6 -trichloro-4-biphenylol; 2 ,3 ,4 ,5 -tetrachloro-4-biphenylol; p-t octylphenol; p-nonylphenol; naringenin; kepone; resveratrol; and 2,2-bis(p hydroxyphenyl)-1,1,1-trichloroethane. The results show that synthetic and phytoestrogens are weakly estrogenic but induce distinct patterns of ER agonist/antagonist activities that are cell context- and promoter-dependent, suggesting that these compounds will induce tissue-specific (in vivo(ER agonist or antagonist activities. These results suggest that other receptors, such as the aryl hydrocarbon receptor, that also bind structurally diverse ligands may exhibit unique responses in vivo that are not predicted by standard in vitro bioassays. PMID- 12634122 TI - Characterization of gene expression changes associated with MNNG, arsenic, or metal mixture treatment in human keratinocytes: application of cDNA microarray technology. AB - The identification of molecular markers related to critical biological processes during carcinogenesis may aid in the evaluation of carcinogenic potentials of chemicals and chemical mixtures. Work from our laboratory demonstrated that a single treatment with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) enhanced spontaneous malignant transformation of the human keratinocyte cell line RHEK-1. In contrast, chronic low-level exposure of cells to arsenic alone or in a mixture containing arsenic, cadmium, chromium, and lead inhibited malignant conversion. To identify changes in gene expression that influence these different outcomes, cDNA microarray technology was used. Analysis of multiple human arrays in MNNG transformed RHEK-1 cells, designated OM3, and those treated with arsenic or the arsenic-containing metal mixture showed unique patterns of gene expression. Genes that were overexpressed in OM3 included oncogenes, cell cycle regulators, and those involved in signal transduction, whereas genes for DNA repair enzymes and inhibitors of transformation and metastasis were suppressed. In arsenic-treated cells, multiple DNA repair proteins were overexpressed. Mixture-treated cells showed increased expression of a variety of genes including metallothioneins and integrin 4. These cells showed decreased expression of oncogenes, DNA repair proteins, and genes involved in the mitogen-activated protein kinase pathway. For comparison we are currently analyzing gene expression changes in RHEK-1 cells transformed by other means. The goal of these studies is to identify common batteries of genes affected by chemical modulators of the carcinogenic process. Mechanistic studies may allow us to correlate alterations in their expression with sequential stages in the carcinogenic process and may aid in the risk assessment of other xenobiotics. PMID- 12634124 TI - Gene induction studies and toxicity of chemical mixtures. AB - As part of its mixtures program, the Agency for Toxic Substances and Disease Registry (ATSDR) supports in vitro and limited in vivo toxicity testing to further our understanding of the toxicity and health effects of chemical mixtures. There are increasing concerns that environmental chemicals adversely affect the health of humans and wildlife. These concerns have been augmented by the realization that exposure to chemicals often occurs to mixtures of these chemicals that may exhibit complex synergistic or antagonistic interactions. To address such concerns, we have conducted two studies with techniques that are being used increasingly in experimental toxicology. In the first study, six organochlorine pesticides (4,4 -DDT, 4,4 -DDD, 4,4 -DDE, aldrin, dieldrin, or endrin) were selected from the ATSDR Comprehensive Environmental Response, Compensation and Liability Act of 1980 (or Superfund) priority list and tested for their ability to modulate transcriptional activation of an estrogen responsive reporter gene in transfected HeLa cells. In these assays, HeLa cells cotransfected with an expression vector encoding estrogen receptor and an estrogen-responsive chloramphenicol acetyltransferase (CAT) reporter plasmid were dosed with and without selected environmental chemicals either individually or in defined combinations. Estradiol consistently elicited 10- to 23-fold dose dependent inductions in this assay. By contrast, all six of the organochlorine pesticides showed no detectable dose-related response when tested either individually or in binary combinations. Thus, these chemicals as binary mixtures do not exhibit any additional estrogenicity at the levels tested in these assays. In the second study, arsenic [As(V)], cadmium [Cd(II)], chromium [Cr(III, VI)], and lead [Pb(II)] were tested in a commercially developed assay system, CAT-Tox (L), to identify metal-responsive promoters and to determine whether the pattern of gene expression changed with a mixture of these metals. This assay employs a battery of recombinant HepG2 cell lines to test the transcriptional activation capacity of xenobiotics in any of 13 different signal-transduction pathways. Singly, As(V), Cd(II), Cr(III, VI), and Pb(II) produced complex induction profiles in these assays. However, no evidence of synergistic activity was detected with a mixture of Cd(II), Cr(III), and Pb(II). These results have shown metal activation of gene expression through several previously unreported signal transduction pathways and thus suggest new directions for future studies into their biochemical mechanisms of toxicity. In conclusion, the (italic)in vitro(/italic) methods used in these studies provide insights into complex interactions that occur in cellular systems and could be used to identify biomarkers of exposure to other environmental chemical mixtures. PMID- 12634125 TI - Application of biologically based computer modeling to simple or complex mixtures. AB - The complexity and the astronomic number of possible chemical mixtures preclude any systematic experimental assessment of toxicology of all potentially troublesome chemical mixtures. Thus, the use of computer modeling and mechanistic toxicology for the development of a predictive tool is a promising approach to deal with chemical mixtures. In the past 15 years or so, physiologically based pharmacokinetic/pharmacodynamic (PBPK/PD) modeling has been applied to the toxicologic interactions of chemical mixtures. This approach is promising for relatively simple chemical mixtures; the most complicated chemical mixtures studied so far using this approach contained five or fewer component chemicals. In this presentation we provide some examples of the utility of PBPK/PD modeling for toxicologic interactions in chemical mixtures. The probability of developing predictive tools for simple mixtures using PBPK/PD modeling is high. Unfortunately, relatively few attempts have been made to develop paradigms to consider the risks posed by very complex chemical mixtures such as gasoline, diesel, tobacco smoke, etc. However, recent collaboration between scientists at Colorado State University and engineers at Rutgers University attempting to use reaction network modeling has created hope for the possible development of a modeling approach with the potential of predicting the outcome of toxicology of complex chemical mixtures. We discuss the applications of reaction network modeling in the context of petroleum refining and its potential for elucidating toxic interactions with mixtures. PMID- 12634126 TI - Evaluating quantitative formulas for dose-response assessment of chemical mixtures. AB - Risk assessment formulas are often distinguished from dose-response models by being rough but necessary. The evaluation of these rough formulas is described here, using the example of mixture risk assessment. Two conditions make the dose response part of mixture risk assessment difficult, lack of data on mixture dose response relationships, and the need to address risk from combinations of chemicals because of public demands and statutory requirements. Consequently, the U.S. Environmental Protection Agency has developed methods for carrying out quantitative dose-response assessment for chemical mixtures that require information only on the toxicity of single chemicals and of chemical pair interactions. These formulas are based on plausible ideas and default parameters but minimal supporting data on whole mixtures. Because of this lack of mixture data, the usual evaluation of accuracy (predicted vs. observed) cannot be performed. Two approaches to the evaluation of such formulas are to consider fundamental biological concepts that support the quantitative formulas (e.g., toxicologic similarity) and to determine how well the proposed method performs under simplifying constraints (e.g., as the toxicologic interactions disappear). These ideas are illustrated using dose addition and two weight-of-evidence formulas for incorporating toxicologic interactions. PMID- 12634128 TI - Experimental designs for mixtures of chemicals along fixed ratio rays. AB - Experimental design is important when studying mixtures/combinations of chemicals. The traditional approach for studying mixtures/combinations of multiple chemicals involves response surface methodology, often supported by factorial designs. Although such an approach permits the investigation of both the effects of individual chemicals and their interactions, the number of design points needed to study the chemical mixtures becomes prohibitive when the number of compounds increases. Fixed ratio ray designs have been developed to reduce the amount of experimental effort when interest can be restricted to a specific ray. We focus on the design and analysis issues involved in studying mixtures/combinations of compounds along fixed ratio rays of the compounds. To obtain the inference regarding the interactions among the compounds, we show that the only data required are those along the fixed ratio ray. PMID- 12634127 TI - Molecular circuits, biological switches, and nonlinear dose-response relationships. AB - Signaling motifs (nuclear transcriptional receptors, kinase/phosphatase cascades, G-coupled protein receptors, etc.) have composite dose-response behaviors in relation to concentrations of protein receptors and endogenous signaling molecules. "Molecular circuits" include the biological components and their interactions that comprise the workings of these signaling motifs. Many of these molecular circuits have nonlinear dose-response behaviors for endogenous ligands and for exogenous toxicants, acting as switches with "all-or-none" responses over a narrow range of concentration. In turn, these biological switches regulate large-scale cellular processes, e.g., commitment to cell division, cell differentiation, and phenotypic alterations. Biologically based dose-response (BBDR) models accounting for these biological switches would improve risk assessment for many nonlinear processes in toxicology. These BBDR models must account for normal control of the signaling motifs and for perturbations by toxic compounds. We describe several of these biological switches, current tools available for constructing BBDR models of these processes, and the potential value of these models in risk assessment. PMID- 12634129 TI - Toxicological evaluation of complex mixtures by pattern recognition: correlating chemical fingerprints to mutagenicity. AB - We describe the use of pattern recognition and multivariate regression in the assessment of complex mixtures by correlating chemical fingerprints to the mutagenicity of the mixtures. Mixtures were 20 organic extracts of exhaust particles, each containing 102-170 individual compounds such as polycyclic aromatic hydrocarbons (PAHs), nitro-PAHs, oxy-PAHs, and saturated hydrocarbons. Mixtures were characterized by full-scan GC-MS (gas chromatography-mass spectrometry). Data were resolved into peaks and spectra for individual compounds by an automated curve resolution procedure. Resolved chromatograms were integrated, resulting in a predictor matrix that was used as input to a principal component analysis to evaluate similarities between mixtures (i.e., classification). Furthermore, partial least-squares projections to latent structures were used to correlate the GC-MS data to mutagenicity, as measured in the Ames Salmonella assay (i.e., calibration). The best model (high r2 and Q2) identifies the variables that co-vary with the observed mutagenicity. These variables may subsequently be identified in more detail. Furthermore, the regression model can be used to predict mutagenicity from GC-MS chromatograms of other organic extracts. We emphasize that both chemical fingerprints as well as detailed data on composition can be used in pattern recognition. PMID- 12634130 TI - Physiological modeling and extrapolation of pharmacokinetic interactions from binary to more complex chemical mixtures. AB - The available data on binary interactions are yet to be considered within the context of mixture risk assessment because of our inability to predict the effect of a third or a fourth chemical in the mixture on the interacting binary pairs. Physiologically based pharmacokinetic (PBPK) models represent a potentially useful framework for predicting the consequences of interactions in mixtures of increasing complexity. This article highlights the conceptual basis and validity of PBPK models for extrapolating the occurrence and magnitude of interactions from binary to more complex chemical mixtures. The methodology involves the development of PBPK models for all mixture components and interconnecting them at the level of the tissue where the interaction is occurring. Once all component models are interconnected at the binary level, the PBPK framework simulates the kinetics of all mixture components, accounting for the interactions occurring at various levels in more complex mixtures. This aspect was validated by comparing the simulations of a binary interaction-based PBPK model with experimental data on the inhalation kinetics of m-xylene, toluene, ethyl benzene, dichloromethane, and benzene in mixtures of varying composition and complexity. The ability to predict the kinetics of chemicals in complex mixtures by accounting for binary interactions alone within a PBPK model is a significant step toward the development of interaction-based risk assessment for chemical mixtures. PMID- 12634131 TI - Molecular identification of organic compounds in atmospheric complex mixtures and relationship to atmospheric chemistry and sources. AB - This article describes a chemical characterization approach for complex organic compound mixtures associated with fine atmospheric particles of diameters less than 2.5 m (PM2.5). It relates molecular- and bulk-level chemical characteristics of the complex mixture to atmospheric chemistry and to emission sources. Overall, the analytical approach describes the organic complex mixtures in terms of a chemical mass balance (CMB). Here, the complex mixture is related to a bulk elemental measurement (total carbon) and is broken down systematically into functional groups and molecular compositions. The CMB and molecular-level information can be used to understand the sources of the atmospheric fine particles through conversion of chromatographic data and by incorporation into receptor-based CMB models. Once described and quantified within a mass balance framework, the chemical profiles for aerosol organic matter can be applied to existing air quality issues. Examples include understanding health effects of PM2.5 and defining and controlling key sources of anthropogenic fine particles. Overall, the organic aerosol compositional data provide chemical information needed for effective PM2.5 management. PMID- 12634132 TI - Biodegradation kinetics of aromatic hydrocarbon mixtures by pure and mixed bacterial cultures. AB - Microbial growth on pollutant mixtures is an important aspect of bioremediation and wastewater treatment. However, efforts to develop mathematical models for mixed substrate kinetics have been limited. Nearly all models group either the microbial population (as "biomass") or the chemical species (e.g., as biological oxygen demand). When individual chemical species are considered, most models assume either no interaction or that the nature of the interaction is competition for the same rate-limiting enzyme. And when individual microbial species are considered, simple competition for the growth substrate is the only interaction included. Here, we present results using Pseudomonas putida F1 and Burkholderia sp. strain JS150 growing individually and together on benzene, toluene, phenol, and their mixtures and compare mathematical models to describe these results. We demonstrate that the simple models do not accurately predict the outcome of these biodegradation experiments, and we describe the development of a new model for substrate mixtures, the sum kinetics with interaction parameters (SKIP) model. In mixed-culture experiments, the interactions between species were substrate dependent and could not be predicted by simple competition models. Together, this set of experimental and modeling results presents our current state of work in this area and identifies challenges for future modeling efforts. PMID- 12634133 TI - Development of a research strategy for integrated technology-based toxicological and chemical evaluation of complex mixtures of drinking water disinfection byproducts. AB - Chemical disinfection of water is a major public health triumph of the 20th century. Dramatic decreases in both morbidity and mortality of waterborne diseases are a direct result of water disinfection. With these important public health benefits comes low-level, chronic exposure to a very large number of disinfection byproducts (DBPs), chemicals formed through reaction of the chemical disinfectant with naturally occurring inorganic and organic material in the source water. This article provides an overview of joint research planning by scientists residing within the various organizations of the U.S. Environmental Protection Agency Office of Research and Development. The purpose is to address concerns related to potential health effects from exposure to DBPs that cannot be addressed directly from toxicological studies of individual DBPs or simple DBP mixtures. Two factors motivate the need for such an investigation of complex mixtures of DBPs: a) a significant amount of the material that makes up the total organic halide and total organic carbon portions of the DBPs has not been identified; and b) epidemiologic data, although not conclusive, are suggestive of potential developmental, reproductive, or carcinogenic health effects in humans exposed to DBPs. The plan is being developed and the experiments necessary to determine the feasibility of its implementation are being conducted by scientists from the National Health and Environmental Effects Research Laboratory, the National Risk Management Research Laboratory, the National Exposure Research Laboratory, and the National Center for Environmental Assessment. PMID- 12634134 TI - BioMOL: a computer-assisted biological modeling tool for complex chemical mixtures and biological processes at the molecular level. AB - A chemical engineering approach for the rigorous construction, solution, and optimization of detailed kinetic models for biological processes is described. This modeling capability addresses the required technical components of detailed kinetic modeling, namely, the modeling of reactant structure and composition, the building of the reaction network, the organization of model parameters, the solution of the kinetic model, and the optimization of the model. Even though this modeling approach has enjoyed successful application in the petroleum industry, its application to biomedical research has just begun. We propose to expand the horizons on classic pharmacokinetics and physiologically based pharmacokinetics (PBPK), where human or animal bodies were often described by a few compartments, by integrating PBPK with reaction network modeling described in this article. If one draws a parallel between an oil refinery, where the application of this modeling approach has been very successful, and a human body, the individual processing units in the oil refinery may be considered equivalent to the vital organs of the human body. Even though the cell or organ may be much more complicated, the complex biochemical reaction networks in each organ may be similarly modeled and linked in much the same way as the modeling of the entire oil refinery through linkage of the individual processing units. The integrated chemical engineering software package described in this article, BioMOL, denotes the biological application of molecular-oriented lumping. BioMOL can build a detailed model in 1-1,000 CPU sec using standard desktop hardware. The models solve and optimize using standard and widely available hardware and software and can be presented in the context of a user-friendly interface. We believe this is an engineering tool with great promise in its application to complex biological reaction networks. PMID- 12634135 TI - Protein binding of isofluorophate in vivo after coexposure to multiple chemicals. AB - Full toxicologic profiles of chemical mixtures, including dose-response extrapolations to realistic exposures, is a prohibitive analytical problem, even for a restricted class of chemicals. We present an approach to probing in vivo interactions of pesticide mixtures at relevant low doses using a monitor compound to report the response of biochemical pathways shared by mixture components. We use accelerator mass spectrometry (AMS) to quantify [14C] diisopropylfluorophosphate as a tracer at attomole levels with 1-5% precision after coexposures to parathion (PTN), permethrin (PER), and pyridostigmine bromide separately and in conjunction. Pyridostigmine shows an overall protective effect against tracer binding in plasma, red blood cells, muscle, and brain that is not explained as competitive protein binding. PTN and PER induce a significant 25-30% increase in the amount of tracer reaching the brain with or without pyridostigmine. The sensitivity of AMS for isotope-labeled tracer compounds can be used to probe the physiologic responses of specific biochemical pathways to multiple compound exposures. PMID- 12634136 TI - Introduction--the methyl parathion story: a chronicle of misuse and preventable human exposure. AB - In the fall of 1994, Lorain County, Ohio, became the site of the first investigation of several large-scale incidences in which the organophosphate pesticide methyl parathion was illegally applied to private residences. The extent of potential human exposure to this pesticide led the Ohio Department of Health to formally request technical assistance from the Centers for Disease Control and Prevention (CDC). This article describes the initial investigation of 64 homes in Ohio and introduces the method of using both biological markers of exposure (p-nitrophenol levels in human urine samples) and environmental markers of contamination in dust and air samples when making public health decisions about the cleanup of homes sprayed with methyl parathion. The results of the CDC rapid investigation led the U.S. Environmental Protection Agency to declare the contaminated homes in Lorain County a Superfund cleanup site. Seven years after the Lorain incident, and after subsequent Superfund actions had been implemented in Illinois and Mississippi, researchers participated in an expanded session devoted to methyl parathion at the 11th Annual Meeting of the International Society of Exposure Analysis held in Charleston, South Carolina, in the fall of 2001. The articles included in this monograph are based on presentations at that meeting. They report previously unpublished data that tell the methyl parathion story from different perspectives, each providing in-depth information about separate aspects of this multistate, multiagency, and multimillion dollar chemical exposure. This monograph is the methyl parathion story. PMID- 12634138 TI - Assessment of human exposure and human health effects after indoor application of methyl parathion in Lorain County, Ohio, 1995-1996. AB - In January 1995 the U.S. Environmental Protection Agency declared methyl parathion-contaminated homes in Lorain County, Ohio, as a Superfund cleanup site. During the 2-year cleanup, the Centers for Disease Control and Prevention in collaboration with county and city health officials conducted a study of exposure and health effects among residents. We administered 254 household and 747 individual questionnaires; urine analysis for p-nitrophenol (PNP, a metabolite of methyl parathion) was available for 626 participants. We also reviewed medical records of 49 people who were hospitalized or died after their homes were sprayed. People living in homes sprayed <180 days previously were most likely to have the highest PNP levels (22.9% > 100 ppb PNP), but even people living in homes sprayed more than a year previously appeared to be highly exposed (8.5% > 100 ppb PNP). The National Health and Nutrition Examination Survey reference range is 0-63 ppb. Median detectable PNP levels among children younger than 3 years of age were 93.9 ppb compared with 41.6 ppb among people older than 3 years. Younger children appeared to be at greatest risk of exposure. In none of the medical records that we reviewed did a health care provider consider pesticide poisoning as a potential etiology. PMID- 12634137 TI - Urinary p-nitrophenol as a biomarker of household exposure to methyl parathion. AB - Methyl parathion (MP) is an organophosphate pesticide illegally applied to the interiors of many hundreds of homes throughout the United States by unlicensed pesticide applicators. Public health authorities developed a protocol for investigating contaminated homes and classifying their need for public health interventions. This protocol included environmental screening for MP contamination and 1-day biomonitoring (a.m. and p.m. spot urine samples) of household members for p-nitrophenol (PNP), a metabolite of MP. The variability of urinary PNP excretion under these exposure conditions was unknown. We collected a.m. and p.m. spot urine samples for 7 consecutive days from 75 individuals, who were members of 20 MP-contaminated households in the greater Chicago, Illinois, area, and analyzed them for PNP. We also assessed the ability of the 1-day sampling protocol to correctly classify exposed individuals and households according to their need for public health interventions, assuming that 1 week of sampling (14 urinary PNPs) represented their true exposure condition. The coefficient of variation of log urinary PNPs for individuals over the course of 7 days of a.m. and p.m. sampling averaged about 15%. Adjusting for urinary excretion of creatinine improved reproducibility of urinary PNPs among children but not among adults. The 1-day protocol correctly classified true risk category in 92% of individuals and 85% of households. The data contained in this study can be used to refine what is already a reasonable and effective approach to identifying MP-exposed households and determining the appropriate public health intervention. PMID- 12634139 TI - An investigation of unexplained infant deaths in houses contaminated with methyl parathion. AB - In Lorain County, Ohio, unexplained infant deaths in homes sprayed with methyl parathion (MP), an organophosphate (OP) pesticide, prompted an investigation to determine whether infants living in treated homes are at higher risk for unexplained death. A case was defined as any death of an infant (12 months of age) in Lorain County between 1 January 1990 and 31 December 1994, attributed to sudden infant death syndrome (SIDS) or other unknown natural causes. For each case infant, birth certificate data were used to identify two control infants matched with regard to date of birth, sex, city of residence, and maternal race and educational level. Wipe samples from the home address listed on the birth certificate of control infants or the death certificate of case infants were analyzed for MP. Birth certificates provided additional risk factor information. The relationship between MP contamination and unexplained death was analyzed by exact conditional logistic regression. Wipe samples were collected from the residences of 34 case infants and 72 control infants. MP (>0.02 mg/100 cm2) was detected in five homes, three of which had been occupied by case infants. Case infants were 4.6 times more likely than control infants to have lived in MP treated homes, but the confidence interval (CI) was wide (95% CI: 0.2, 274.7) and included 1. Maternal smoking, young maternal age, and the presence of other siblings in the family were each independently predictive of case status. In a multivariate model adjusting for these other variables and the matching variables, the estimated risk associated with MP exposure was 13.0 (95% CI: 0.2, 2,685.0). Although this association was not statistically significant and should be interpreted cautiously, it suggests an increased risk for unexplained death among infants living in MP-contaminated homes. The relationship between children's health and exposure to OP pesticides including MP should be evaluated further. PMID- 12634140 TI - Public health decisions: the laboratory's role in the Lorain County, Ohio, investigation. AB - In 1994 officials from the Ohio Department of Health reported that some residents of Lorain County, Ohio, possibly had been exposed to methyl parathion (MP), a highly toxic restricted-use pesticide. The U.S. Centers for Disease Control and Prevention (CDC) assisted in the investigation by providing epidemiologic and laboratory support to the state and local health departments. Although the initial investigation found MP inside the homes, it was unclear if the residents were exposed. CDC used a new biological monitoring method to measure urinary p nitrophenol (PNP), the metabolite of MP. This biological monitoring measures the internal dose from exposure to toxic chemicals from all routes. Laboratory analyses demonstrated that the urine of residents contained moderate to high levels of PNP, with median, mean, and highest reported concentrations of 28, 240, and 4,800 g/L, respectively, thus confirming exposure of the residents. Almost 80% of the residents had urinary PNP concentrations above the 95th percentile of the reference range concentrations. This information, combined with other analytical results of air and wipe tests, guided public health officials' decisions about the potential risk in each household. In this article we illustrate the laboratory's role in providing information to assist in making these public health decisions. Furthermore, it illustrates how a multidisciplinary team from various governmental agencies worked together to protect the public's health. PMID- 12634141 TI - Methyl parathion in residential properties: relocation and decontamination methodology. AB - In November 1994 methyl parathion (MP), a restricted agricultural pesticide, was discovered to have been illegally sprayed within hundreds of residences in Lorain County, Ohio. Surface levels and air concentrations of MP revealed detectable levels of the pesticide 3 years after spraying. Because of the high toxicity of MP (lethal dose to 50% of rats tested [LD50] = 15 mg/kg) and long half-life indoors, risk-based relocation and decontamination criteria were created. Relocation criteria were derived based on levels of p-nitrophenol in urine, a metabolic byproduct of MP exposure. In Ohio, concentrations of MP on surfaces and in the air were also used to trigger relocations. The criteria applied in Ohio underwent refinement as cases of MP misuse were found in Mississippi and then in several other states. The MP investigation (1994-1997) was the largest pesticide misuse case in the nation, ultimately involving the sampling of 9,000 residences and the decontamination of 1,000 properties. This article describes the methodology used for relocation of residents and decontamination of properties having MP. PMID- 12634142 TI - Analysis of environmental and biologic methyl parathion data to improve future data collection. AB - The Agency for Toxic Substances and Disease Registry analyzed concurrently collected data on environmental methyl parathion (MP) and urinary p-nitrophenol (PNP) at the request of the U.S. Environmental Protection Agency (U.S. EPA). The purpose of the analysis was to assess whether individuals' age or level of residential MP contamination might predict their urinary PNP level. Unlicensed pesticide applicators had sprayed residences in Mississippi with MP, which is approved as a pesticide only for outdoor agricultural use. Data were received from Mississippi for MP wipe sample levels for 409 homes and urinary PNP levels for 929 residents of the residences sampled. In addition to descriptive and bivariate analyses, ordinal logistic regression was performed after categorizing the data. Interpretation of results was limited by several identified data gaps and pre-existing data-quality issues. On the basis of the lessons learned from identified data gaps, specific recommendations were made to the U.S. EPA for improving future data collection methods for more meaningful exposure assessment in similar environmental contaminations. The recommended changes were successfully incorporated in subsequent data collected by other states that had experienced similar residential MP spraying. PMID- 12634143 TI - Chicago area methyl parathion response. AB - The Illinois Department of Public Health participated in the Chicago, Illinois, area methyl parathion (MP) response with several other federal, state, and local government agencies beginning in April 1997. This response was initiated on evidence that hundreds of homes in the Chicago area were illegally treated for cockroaches with MP over a period of several years. Through applicator receipt books and information reported by property owners and tenants, 968 homes were identified as having been treated with MP. Upon implementation of a response plan developed by the Methyl Parathion Health Sciences Steering Committee, environmental sampling and urine monitoring were provided for eligible households. Environmental sampling was conducted in 903 homes, with MP detected above levels of concern in 596 residences. Residents of these homes were offered urine sampling to determine the extent of exposure to MP. Urine samples were collected and analyzed for p-nitrophenol in 1,913 individuals. Implementation of the protocol resulted in 550 residents being relocated during the remediation of 100 households. PMID- 12634144 TI - Methods of assessing neurobehavioral development in children exposed to methyl parathion in Mississippi and Ohio. AB - Methyl parathion (MP), an organophosphate pesticide, was sprayed illegally for pest control in U.S. residences and businesses in Mississippi and Ohio. To evaluate the association between MP exposure and neurobehavioral development, children 6 years of age or younger at the time of the spraying and local comparison groups of unexposed children were assessed using the pediatric environmental neurobehavioral test battery (PENTB). The PENTB is composed of informant-based procedures (parent interview and questionnaires) and performance based procedures (neurobehavioral tests for children 4 years of age or older) that evaluate each of the four broad domains (cognitive, motor, sensory, and affect) essential to neurobehavioral assessment. Children were classified as exposed or unexposed using urinary p-nitrophenol (PNP) levels and environmental wipe samples for MP. Exposure was defined as a urinary PNP level of greater than or equal to 100 ppb for the child or any other individual living in the household. Environmental wipe sample levels of greater than or equal to 150 g MP/100 cm2 and greater than or equal to 132.9 g MP/100 cm2 were used to define MP exposure for children living in Mississippi and Ohio, respectively. The PENTB was first administered in summer 1999 (year 1). The PENTB was readministered in summer 2000 (year 2) to children who participated in year 1 of the study. A description of the methods used in the study are presented. Results of data analyses for both years of the study will be presented in a separate publication. PMID- 12634200 TI - The information technology age is dawning for medical education. PMID- 12634145 TI - Measurement of p-nitrophenol in the urine of residents whose homes were contaminated with methyl parathion. AB - During the last several years, illegal commercial application of methyl parathion (MP) in domestic settings in several U.S. Southeastern and Midwestern States has affected largely inner-city residents. As part of a multiagency response involving the U.S. Environmental Protection Agency (U.S. EPA), the Agency for Toxic Substances and Disease Registry (ATSDR), and state and local health departments, our laboratory developed a rapid, high-throughput, selective method for quantifying p-nitrophenol (PNP), a biomarker of MP exposure, using isotope dilution high-performance liquid chromatography-tandem mass spectrometry. We measured PNP in approximately 16,000 samples collected from residents of seven different states. Using this method, we were able to receive sample batches from each state; prepare, analyze, and quantify the samples for PNP; verify the results; and report the data to the health departments and ATSDR in about 48 hr. These data indicate that many residents had urinary PNP concentrations well in excess of those of the general U.S. population. In fact, their urinary PNP concentrations were more consistent with those seen in occupational settings or in poisoning cases. Although these data, when coupled with other MP metabolite data, suggest that many residents with the highest concentrations of urinary PNP had significant exposure to MP, they do not unequivocally rule out exposure to PNP resulting from environmental degradation of MP. Even with their limitations, these data were used with the assumption that all PNP was derived from MP exposure, which enabled the U.S. EPA and ATSDR to develop a comprehensive, biologically driven response that was protective of human health, especially susceptible populations, and included clinical evaluations, outreach activities, community education, integrated pest management, and decontamination of homes. PMID- 12634201 TI - Introducing HEAL: the Health Education Assets Library. AB - Digital multimedia, such as images and videos, are playing an increasingly important role in health sciences education. Educators, however, often do not have the time or resources to create high-quality materials. The authors describe the development of a new Health Education Assets Library (HEAL), a freely accessible, national library of high-quality digital multimedia to support all levels of health sciences education. HEAL's primary mission is to provide educators with high-quality and free multimedia materials (such as images and videos) to augment health science education. In addition, HEAL is working with other organizations to establish a network of distributed databases of high quality teaching resources. By using state-of-the-art Internet technologies HEAL enables educators across the country to efficiently search and retrieve teaching materials from a variety of sources. PMID- 12634202 TI - Tufts Health Sciences Database: lessons, issues, and opportunities. AB - The authors present their seven-year experience with developing the Tufts Health Sciences Database (Tufts HSDB), a database-driven information management system that combines the strengths of a digital library, content delivery tools, and curriculum management. They describe a future where online tools will provide a health sciences learning infrastructure that fosters the work of an increasingly interdisciplinary community of learners and allows content to be shared across institutions as well as with academic and commercial information repositories. The authors note the key partners in Tufts HSDB's success--the close collaboration of the health sciences library, educational affairs, and information technology staff. Tufts HSDB moved quickly from serving the medical curriculum to supporting Tufts' veterinary, dental, biomedical sciences, and nutrition schools, thus leveraging Tufts HSDB research and development with university-wide efforts including Internet2 middleware, wireless access, information security, and digital libraries. The authors identify major effects on teaching and learning, e.g., what is better taught with multimedia, how faculty preparation and student learning time can be more efficient and effective, how content integration for interdisciplinary teaching and learning is promoted, and how continuous improvement methods can be integrated. Also addressed are issues of faculty development, copyright and intellectual property, budgetary concerns, and coordinating IT across schools and hospitals. The authors describe Tufts' recent experience with sharing its infrastructure with other schools, and welcome inquiries from those wishing to explore national and international partnerships to create a truly open and integrated infrastructure for education across the health sciences. PMID- 12634204 TI - Development of an undergraduate medical curriculum: the McGill experience. AB - In 1997 the Faculty of Medicine at McGill University received a grant from the Molson Foundation. The primary project deliverable, which the authors describe, was an online, multimedia-enhanced, undergraduate medical curriculum. The decision to develop an electronic curriculum was predicated on the belief that the integration of educational technology within mainstream material delivered a "value added" component to both students and faculty, which would, in turn, facilitate teaching and learning. Pedagogical values were deemed to include: (1) the ability to use the media to implement adult learning principles such as learner-centered, self-directed and guided learning, (2) the inherent interactivity of the technology, (3) the potential of the technology to provide a powerful means for fostering forms of "termless" learning that students will need to practice medicine, (4) recognition that use of multimedia can address, in part, the variety of learning styles evidenced by students in the lecture hall and classroom, and (5) the provision of opportunities for horizontal and vertical curricular integration. In addition, it was anticipated that an electronic curriculum would permit: (1) easy incorporation of informatics within mainstream curricula, (2) centralization and standardization of curricular material, (3) editorial functionality for revisions and updates, (4) wide accessibility of material irrespective of venue, (5) search functionality for faculty and students, (6) the ability to perform curriculum inventory, and (7) the potential for use to compensate for decreased faculty time. The ongoing experience at McGill has shown that the merging of technology and pedagogy requires a substantial commitment of resources and recognition of faculty time and change management issues. PMID- 12634206 TI - A Web-based compendium of clinical questions and medical evidence to educate internal medicine residents. AB - The authors designed an electronic database of clinical questions (CQs) and medical evidence and implemented it in 2001-02 at Duke University Medical Center and the Veterans Administration Medical Center in Durham, North Carolina. This Web-based data collection system is called the Critical Appraisal Resource (CAR) and is still in operation. This report is of ten months of the system's operation. During their medicine ward rotations, residents entered CQs into the CAR; they also entered Medline reference links and validated article summaries. Residents' utilization of the CAR database, Medline, and other electronic resources was prospectively measured. In addition, residents were prospectively surveyed regarding the impact of each question and associated reference on medical decision making for individual patients. Over ten months, residents entered 625 patient-based CQs into the CAR and were able to obtain useful information from the medical literature on 82% of the CQs they searched. The two most prevalent CQ types were therapy and diagnosis questions (53% and 22%). Sixty percent of the therapy articles considered useful were reports of randomized controlled trials. Residents obtained 77% of their useful data from Medline. They reported that obtaining useful data altered patient management 47% of the time. Residents used the CAR as a resource, searching the database for information 1,035 times over the study period. In summary, the use of an evidence-based critical appraisal resource led residents to engage the medical literature on behalf of their patients and influenced approximately half of their patient-care decisions. Residents benefited from questions previously searched by other residents, allowing them to address a wider spectrum of CQs during ward rotations. PMID- 12634207 TI - "You're not doing my surgery, are you?". PMID- 12634208 TI - CurrMIT: a tool for managing medical school curricula. AB - The AAMC Curriculum Management & Information Tool (CurrMIT) is a relational database containing curriculum information from medical schools throughout the United States and Canada. CurrMIT can be used to document details of instruction, such as outcome objectives, resources, content, educational methods, assessment methods, and educational sites, which are being employed in curricula. CurrMIT contains basic information about nearly all required courses and clerkships being offered in the United States and Canada. The database contains descriptions of more than 15,000 courses and clerkships; approximately 115,000 "sessions"--e.g., lectures, labs, small-group discussions--and more than 400,000 keywords and word strings documenting the specific details of instruction associated with the courses, clerkships, and sessions. Some specific uses that schools have made of CurrMIT include review of demographics among patient cases being used in a case based curriculum; comparisons of educational experiences between two geographically separate clinical campuses; and identification of unplanned redundancies and gaps in curricular content. CurrMIT has been designed to accommodate data from virtually any medical school curriculum; "traditional 2+2" curricula, problem-based curricula, and systems-based curricula, and variations of each of these, have been entered in CurrMIT by medical schools. The authors give an overview of the technology upon which the system is built and the training materials and workshops that the AAMC provides to faculty to support CurrMIT's use, and end by describing enhancements being planned for the system. PMID- 12634209 TI - Exploring and embracing complexity in a distance-learning curriculum for physicians. AB - The recent pressures on clinical medicine such as the attention to medical error and the challenges of interdisciplinary care have also exerted pressure on health professions education. Educators must now gauge how to redesign education systems to adapt quickly to these disruptions. Sometimes disruptions can be self inflicted, such as the VA National Quality Scholars Fellowship's decision to use interactive video (IV) as its primary medium for delivering the curriculum to its six sites around the nation. The authors describe how this disruption to their education system helped to fashion a learning environment that is adaptable. Along the journey from a classroom-based curriculum to an IV-based curriculum, the authors and others involved in the program learned the basic tenets of IV sessions, redefined the roles of the teachers and learners, and discovered an IV environment that functions as a complex adaptive learning system. This distance learning curriculum can be a model for other health professions education, since it starts with simple rules, changes from within, has a tolerance for unpredictability, and continually moves forward and transforms itself despite tension. PMID- 12634210 TI - Sharing digital teaching resources: breaking down barriers by addressing the concerns of faculty members. AB - PURPOSE: Regional and institutional databases have been created to improve access to educational resources and to avert unnecessary duplication. The growth and success of these databases depend upon the willingness of faculty members to contribute their materials. This qualitative study seeks to identify the barriers that block the free exchange of educational multimedia and the incentives that could be created to overcome educators' concerns. METHOD: In 2001, 34 faculty members from 13 U.S. medical schools each participated in one of five focus groups. They responded to three hypothetical scenarios depicting opportunities for sharing educational resources. Participants' responses were categorized hierarchically and sent back to them for feedback. RESULTS: Participants strongly supported multimedia databases, particularly those that serve a national audience. Obstacles for contributing materials included the lack of institutional recognition for educational innovation, confusing intellectual property policies, the hassle involved in sharing materials, and the perceived commercial potential of the materials. Peer review of educational materials was seen as an important incentive. CONCLUSIONS: Medical schools could benefit from the free exchange of high-quality educational multimedia but need to address the concerns of faculty by clarifying institutional copyright rules, streamlining the donation process, and providing assistance with cataloging assets. Removing departmental pressures to commercialize materials and recognizing peer-reviewed donations as academic achievement could foster a culture of sharing. PMID- 12634211 TI - Integrating Web-based computer cases into a required clerkship: development and evaluation. AB - PURPOSE: To evaluate a five-year experience (1995-2000) developing and integrating computer cases into a required clerkship. METHOD: In Study 1, 54 volunteer students were randomly assigned to study articles, a paper case, or a computer case on low back pain/kidney stones. Students were given an exam immediately after the exercise and one week later. In Study 2, 325 clerkship students were asked to select and complete two computer cases or to prepare assignments on unrelated topics. Among the cases offered were two test cases on low back pain/kidney stones and pneumonia. Questions specific to the computer test cases (CC) and other noncomputer cases (NCC) were in the final exam. Exam scores related to CC questions and NCC questions were compared between the groups of students who did and did not complete the computer cases. Students also rated the computer cases on a questionnaire. RESULTS: In Study 1, reading articles required the most time and received the most negative comments. The students who completed the computer case scored the best on the exam one week later. In Study 2, the students who completed the two computer cases scored significantly higher on the CC questions than did students who studied only one or none of the computer cases (p <.001). There was no difference among groups for the NCC scores (p =.76). Students rated the computer cases with a mean of 6 on a seven-point scale (7 = strongly agree). CONCLUSION: Computer cases are effective learning tools, are well-received by students, and can be successfully integrated into existing clerkships. PMID- 12634213 TI - The reorganization of basic science departments in U.S. medical schools, 1980 1999. AB - The evolution of biomedical science and technology over the last 50 years has made biomedical research inherently interdisciplinary. Such changes have led observers to speculate about the ways in which traditional basic science departments in U.S. medical schools are being changed or consolidated. The authors describe their findings from a study that constructed a 20-year longitudinal database (1980-1999) to examine how basic science departments have been reorganized at U.S. medical schools. The data reveal that, in fact, there were fewer basic science departments in the traditional disciplines of anatomy, biochemistry, microbiology, pharmacology, and physiology in 1999 than in 1980. But as biomedical science has developed in an interdisciplinary manner, new basic science departments have been added. The most frequent type of change, however, has been in the renaming of existing departments. Overall, there were more, not fewer, basic science departments and more, not fewer, faculty members in these departments. These changes, taken together with the growth of interdisciplinary research centers and institutes and changing patterns of biomedical PhD training, affect both teaching and research in academic medicine. First, basic scientists are becoming increasingly dissociated from the traditional disciplines around which medical students' education is often organized. Second, the organization of biomedical research is in a state of transition that is responding to advances in scientific knowledge, technology, and targets of opportunity. PMID- 12634214 TI - Reflective practice and palliative care education: a clerkship responds to the informal and hidden curricula. AB - The authors discuss the damaging influence of informal and hidden curricula on medical students and describe a two-week clerkship in palliative care and clinical ethics at their school (Weill Medical College of Cornell University). This required clerkship, begun in 1999, uses reflective practice and a special pedagogic technique, participant observation, to counteract the influences of the informal and hidden curricula. This technique seeks to immerse the participant observer in the context of care. In their role as participant observers, students are relieved of any direct clinical responsibilities for two weeks so they have time for the careful observation and reflection required and also can consider the humanistic dimensions of practice, which are often displaced by the need to master diagnostic and therapeutic skills. Course objectives include identifying psychosocial and contextual factors that influence care, principles of pain and symptom management, and ethical and legal issues at the end of life. Students are expected to learn how to apply ethical norms to patient care, describe methods of pain and symptom management, communicate in an effective and humanistic manner, and articulate models of patient-centered advocacy. The clerkship fosters professionalism in patient care, appreciation of cultural diversity, and the student's ability to assume responsibility for developing competency in these areas. Although it is too early to know whether this clerkship will ultimately affect the practice patterns of students who experience it, short-term evaluation has been very favorable. PMID- 12634215 TI - Assessing personal qualities in medical school admissions. AB - The authors analyze the challenges to using academic measures (MCAT scores and GPAs) as thresholds for admissions and, for applicants exceeding the threshold, using personal qualities for admission decisions; review the literature on using the medical school interview and other admission data to assess personal qualities of applicants; identify challenges of developing better methods of assessing personal qualities; and propose a unified system for assessment. The authors discuss three challenges to using the threshold approach: institutional self-interest, inertia, and philosophical and historical factors. Institutional self-interest arises from the potential for admitting students with lower academic credentials, which could negatively influence indicators used to rank medical schools. Inertia can make introducing a new system complex. Philosophical and historical factors are those that tend to value maximizing academic measures. The literature identifies up to 87 different personal qualities relevant to the practice of medicine, and selecting the most salient of these that can be practically measured is a challenging task. The challenges to developing better personal quality measures include selecting and operationally defining the most important qualities, measuring the qualities in a cost-effective manner, and overcoming "cunning" adversaries who, with the incentive and resourcefulness, can potentially invalidate such measures. The authors discuss potential methods of measuring personal qualities and propose a unified system of assessment that would pool resources from certification and recertification efforts to develop competencies across the continuum with a dynamic, integrated approach to assessment. PMID- 12634216 TI - "Clinical reasoning theater": a new approach to clinical reasoning education. AB - Recently the authors developed and implemented a new approach to clinical reasoning education called clinical reasoning theater (CRT). The purpose of CRT is to demonstrate the process of clinical reasoning to students through the conversation of a doctor with his or her patient. With students as the audience, the doctor's clinical reasoning skills are modeled in CRT when he or she thinks aloud during conversations with the patient, which makes clear why he or she asks particular questions. In CRT the interaction between doctor and patient is analyzed in such a way that the repeated cycles of questions and answers are revealed to the students. CRT's format and the activities of the three "actors,"(the doctor, the patient, and the audience of students) are described. Two "acts" are repeatedly performed. The first act (the communication between the doctor and the patient) deals with the history taking, physical examination, etc. The second act, the time-out period, is the most important component of CRT. In this act, the doctor explains his or her reflections and discusses suggestions from the audience for additional questions. The preliminary results of students' evaluations of the relevance of CRT reveal that they appreciate CRT. Further, the CRT format offers instructors with previous experience as lecturers only sufficient tools to apply the new approach successfully. PMID- 12634217 TI - Heroics. PMID- 12634219 TI - "Having the right chemistry": a qualitative study of mentoring in academic medicine. AB - PURPOSE: To develop a deeper understanding of mentoring by exploring lived experiences of academic medicine faculty members. Mentoring relationships are key to developing productive careers in academic medicine, but such alliances hold a certain "mystery." METHOD: Using qualitative techniques, between November 1999 and March 2000, the authors conducted individual telephone interviews of 16 faculty members about their experiences with mentoring. Interviews were taped and transcribed and authors identified major themes through multiple readings. A consensus taxonomy for classifying content evolved from comparisons of coding by four reviewers. Themes expressed by participants were studied for patterns of connection and grouped into broader categories. RESULTS: Almost 98% of participants identified lack of mentoring as the first (42%) or second (56%) most important factor hindering career progress in academic medicine. Finding a suitable mentor requires effort and persistence. Effective mentoring necessitates a certain chemistry for an appropriate interpersonal match. Prized mentors have "clout," knowledge, and interest in the mentees, and provide both professional and personal support. In cross-gender mentoring, maintaining clear boundaries is essential for an effective relationship. Same-gender or same-race matches between mentor and mentee were not felt to be essential. CONCLUSIONS: Having a mentor is critical to having a successful career in academic medicine. Mentees need to be diligent in seeking out these relationships and institutions need to encourage and value the work of mentors. Participants without formalized mentoring relationships should look to peers and colleagues for assistance in navigating the academic system. PMID- 12634220 TI - MD/MBA programs in the United States: evidence of a change in health care leadership. AB - PURPOSE: Managerial sciences are playing an increasingly prominent role in the organization and delivery of health care. Despite popular media reports that a rising number of physicians are acquiring a background in this discipline through MD/MBA (medical and master of business administration) programs, no recent study has verified this. This study measured changes in the number and nature of the affiliations between management and medicine in the form of MD/MBA programs in the United States. METHOD: Surveys of admission officers of 125 U.S. allopathic medical schools and of the overseers of each joint MD/MBA degree program were administered in May-October 2001. Main outcome measures included program growth, curriculum and degree requirements, application and admission requirements, and program leadership and organization. RESULTS: The number of MD/MBA programs grew from six to 33 between 1993 and 2001, and 17 more medical schools were considering establishing the joint-degree program. Ten, 15, and 20 programs produced 27, 42, and 61 graduates in 1999, 2000, and 2001, respectively, and over 100 students were expected to graduate per year when all 33 programs matured. Program structures and oversight indicate a spectrum of philosophies regarding the appropriate level of integration of the two degrees. MD/MBA programs apparently attempt to complement medical education with management education rather than the converse. CONCLUSIONS: The growth in the numbers of MD/MBA programs and participants indicates rising cooperation between medical and business schools and increasing interest in management education early in the careers of graduating physicians. PMID- 12634222 TI - Educational effects of international health electives on U.S. and Canadian medical students and residents: a literature review. AB - PURPOSE: To evaluate the educational effects of international health electives (IHEs) on participants. IHEs are a popular component of many medical school and residency program curricula, and are reported to provide benefits in knowledge, skills, and attitudes. METHOD: The authors reviewed all studies reported in Medline and ERIC databases that have assessed the educational effects of IHEs on U.S. and Canadian medical students and residents. Data extracted from eligible studies included type and duration of IHE, details of survey instrument, response rate, comparison group, and outcomes. Seven of the eight eligible studies assessed educational effects on participants using self-reported questionnaires; a single study used an objective measurement of knowledge. RESULTS: Eight studies involving 522 medical students and 166 residents met inclusion criteria. IHEs appear to be associated with career choices in underserved or primary care settings and recruitment to residency programs. They also appear to have positive effects on participants' clinical skills, certain attitudes, and knowledge of tropical medicine. CONCLUSION: IHEs appear to have positive educational influences on participants' knowledge, skills, and attitudes. Furthermore, IHEs may play some role both in recruiting residents and in their choices of careers in primary care and underserved settings. Future directions for research in this field are discussed. PMID- 12634223 TI - Understanding disability. PMID- 12634224 TI - Cyclosporin A monotherapy versus cyclosporin A and methotrexate combination therapy in patients with early rheumatoid arthritis: a double blind randomised placebo controlled trial. AB - OBJECTIVE: To compare the efficacy and toxicity of cyclosporin A (CsA) monotherapy with CsA plus methotrexate (MTX) combination therapy in patients with early rheumatoid arthritis (RA). PATIENTS AND METHODS: 120 patients with active RA, rheumatoid factor positive and/or erosive, were randomly allocated to receive CsA with MTX (n=60) or CsA with placebo (n=60). Treatment with CsA was started in all patients at 2.5 mg/kg/day and increased to a maximum of 5 mg/kg/day in 16 weeks. MTX was started at 7.5 mg/week and increased to a maximal dose of 15 mg/week at week 16. Primary outcomes were clinical remission (Pinals criteria) and radiological damage (Larsen score), at week 48. RESULTS: Treatment was discontinued prematurely in 27 patients in the monotherapy group (21 because of inefficacy, and six because of toxicity) and in 26 patients in the combination therapy group (14 and 12, respectively). At week 48, clinical remission was achieved in four patients in the monotherapy group and in six patients in the combination therapy group (p=0.5). The median Larsen score increased to 10 (25th, 75th centiles: 3.5; 13.3) points in the monotherapy group and to 4 (1.0; 10.5) points in the combination therapy group (p=0.004). 28/60 (47%) of patients in the monotherapy group v 34/60 (57%) of patients in the combination therapy group had reached an American college of Rheumatology 20% (ACR20) response (p=0.36) at week 48; 15/60 (25%) v 29/60 (48%) of patients had reached an ACR50 response (p=0.013); and 7 (12%) v 12 (20%) of patients had reached an ACR70 response (p=0.11). Their was a tendency towards more toxicity in the combination therapy group. CONCLUSIONS: In patients with early RA, neither CsA plus MTX combination therapy nor CsA monotherapy is very effective in inducing clinical remission. Combination therapy is probably better at improving clinical disease activity, and definitely better at slowing radiological progression. Combination therapy should still be compared with methotrexate monotherapy. PMID- 12634226 TI - Microarchitecture and protective mechanisms in synovial tissue from clinically and arthroscopically normal knee joints. AB - BACKGROUND: Synovial biopsies are used to study synovial immunopathology and are increasingly applied for the evaluation of new therapeutic strategies in chronic arthritis. Therefore, it is essential to be informed on the complete spectrum of synovial immunopathology. OBJECTIVE: To describe the cellular content, cytokine and cell adhesion molecule expression in synovial tissue from clinically and arthroscopically normal knees. METHODS: Synovial tissue was obtained from 20 normal subjects at the time of knee joint arthroscopy for unexplained knee pain. Tissue sections were studied for basic histopathology and for a range of cell surface markers, cytokines, and cell adhesion molecules by immunoperoxidase staining. Stained sections were evaluated by semiquantitative scoring and digital image analysis. RESULTS: Normal synovial tissue is composed predominantly of fibrofatty areolar tissue, with a variable thickness of intimal lining, composed of both CD68 positive macrophages and CD55 positive fibroblast-like synoviocytes. Interleukin 1 receptor antagonist (IL1Ra) was frequently detected in the synovial membrane of normal subjects (mean (SD) integrated optical density (IOD)=3809.6 (3893.9)), but both tumour necrosis factor alpha (TNFalpha) and interleukin 1beta (IL1beta) were rarely detected. In addition, cell adhesion molecules were rarely detected in the normal synovial membrane, with the exception of intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Osteoprotegerin (OPG) expression was abundant on synovial lining macrophages (mean (SD) IOD=5276 (4716) as well as endothelial cells (mean (SD) IOD=557 (226)), but receptor activator of nuclear factor kappa ligand (RANKL) expression was rarely seen. CONCLUSIONS: The normal synovial membrane has a variable architecture, including thickness of the lining and the subintimal cell infiltrate, with little inflammatory cytokine production or expression of cell adhesion molecules. The excess of OPG expression over RANKL and IL1Ra over IL1 may be important for protection against joint damage PMID- 12634225 TI - Anti-inflammatory effects of leflunomide on cultured synovial macrophages from patients with rheumatoid arthritis. AB - BACKGROUND: Leflunomide and its active metabolite A77 1726 reversibly inhibits the enzyme dihydro-orotate dehydrogenase, the rate limiting step in de novo synthesis of pyrimidines and progression of the cell cycle in different cell lines, mainly activated T lymphocytes. OBJECTIVE: To analyse in vitro the possible anti-inflammatory effects exerted by A77 1726, on cultured macrophages, obtained from the synovial tissues of patients with rheumatoid arthritis (RA). METHODS: The effects of different doses of A77 1726 on intracytoplasmic expression and extracellular concentration of inflammatory cytokines (tumour necrosis factor alpha (TNFalpha), interleukin (IL) 1beta, IL6), as well as the influence on production and expression of intercellular adhesion molecule-1 (ICAM 1) and cyclo-oxygenase 2 (COX-2) by primary cultures of synovial macrophages from patients with RA, were evaluated by immunocytochemistry and western blot analysis. The observations were made at four and 24 hours. RESULTS: A progressive and significant time and dose dependent decrease of the number of positive macrophages for intracellular TNFalpha and IL1beta, treated with different doses of A77 1726, was found in comparison with untreated cells. The extracellular concentration of TNFalpha was found to be significantly decreased in media containing cultured macrophages at 24 hours for all tested doses of A77 1726. At 24 hours, a significant time and dose dependent decrease of ICAM-1 and COX-2 expression by cultured macrophages after A77 1726 treatment was found. CONCLUSIONS: In conclusion, the mechanism of antiproliferative activity exerted by leflunomide on activated T lymphocytes seems to be the same mechanism (alteration of the cell cycle progression) which interferes with the functions of other activated cells-namely, the monocytes/macrophages, which are strongly involved in the inflammatory reaction in RA synovial tissue. The positive clinical results seem to confirm that leflunomide exerts an anti-inflammatory action on phagocytic cells in short and long term treatment of RA. PMID- 12634227 TI - Cost effectiveness and cost utility analysis of multidisciplinary care in patients with rheumatoid arthritis: a randomised comparison of clinical nurse specialist care, inpatient team care, and day patient team care. AB - OBJECTIVE: To assess the relative cost effectiveness of clinical nurse specialist care, inpatient team care, and day patient team care. METHODS: Incremental cost effectiveness analysis and cost utility analysis, alongside a prospective randomised controlled trial with two year follow up. Included were patients with rheumatoid arthritis (RA) with increasing difficulty in performing activities of daily living over the previous six weeks. Quality of life and utility were assessed by the Rheumatoid Arthritis Quality of Life questionnaire, the Short Form-6D, a transformed rating scale, and the time tradeoff. A cost-price analysis was conducted to estimate the costs of inpatient and day patient hospitalisations. Other healthcare and non-healthcare costs were estimated from cost questionnaires. RESULTS: 210 patients with RA (75% female, median age 59 years) were included. Aggregated over the two year follow up period, no significant differences were found on the quality of life and utility instruments. The costs of the initial treatment were estimated at euro 200 for clinical nurse specialist care, euro 5000 for inpatient team care, and euro 4100 for day patient team care. Other healthcare costs and non-healthcare costs were not significantly different. The total societal costs did not differ significantly between inpatients and day patients, but were significantly lower for the clinical nurse specialist patients by at least euro 5400. CONCLUSIONS: Compared with inpatient and day patient team care, clinical nurse specialist care was shown to provide equivalent quality of life and utility, at lower costs. Therefore, for patients with health conditions that allow for any of the three types of care, the preferred treatment from a health-economic perspective is the care provided by the clinical nurse specialist. PMID- 12634229 TI - Number needed to treat (NNT): implication in rheumatology clinical practice. AB - OBJECTIVE: To calculate the number needed to treat (NNT) and number needed to harm (NNH) from the data in rheumatology clinical trials and systematic reviews. METHODS: The NNTs for the clinically important outcome measures in the rheumatology systematic reviews from the Cochrane Library, issue 2, 2000 and in the original randomised, double blind, controlled trials were calculated. The measure used for calculating the NNT in rheumatoid arthritis (RA) interventions was the American College of Rheumatology 20% improvement or Paulus criteria; in osteoarthritis (OA) interventions, the improvement of pain; and in systemic sclerosis (SSc) interventions, the improvement of Raynaud's phenomenon. The NNH was calculated from the rate of withdrawals due to adverse events from the treatment. RESULTS: The data required for the calculation of the NNT were available in 15 systematic reviews and 11 original articles. For RA interventions, etanercept treatment for six months had the smallest NNT (1.6; 95% confidence interval (CI) 1.4 to 2.0), whereas leflunomide had the largest NNH (9.6; 95% CI 6.8 to 16.7). For OA treatment options, only etodolac and tenoxicam produced significant pain relief compared with placebo (NNT=4.4; 95% CI 2.4 to 24.4 and 3.8; 95% CI 2.5 to 7.3, respectively). For SSc interventions, none were shown to be efficacious in improving Raynaud's phenomenon because the 95% CI of the NNT was infinite. CONCLUSIONS: The NNT and NNH are helpful for clinicians, enabling them to translate the results from clinical trials and systematic reviews to use in routine clinical practice. Both NNT and NNH should be accompanied by a limited 95% CI and adjusted for the individual subject's baseline risk. PMID- 12634230 TI - Hip pain onset in relation to cumulative workplace and leisure time mechanical load: a population based case-control study. AB - OBJECTIVE: In an unselected community sample of adults, to assess the role and importance of exposure to mechanical factors both at work and leisure in the aetiology of hip pain. METHOD: A population based prevalence case-control study. Cases and controls were identified from a population survey of 3847 subjects registered with two general practices in Cheshire, United Kingdom. All subjects received a postal questionnaire which inquired about hip pain during the past month. An occupational history was obtained, including exposure to each of seven physical demands. Information was also obtained on history of participation in eight common sporting activities. RESULTS: 88% of those invited to participate returned a completed questionnaire. The 352 subjects with hip pain were designated as cases, and the remaining 3002 subjects as controls. In people ever employed, hip pain was significantly associated with high cumulative workplace exposure (before onset) of walking long distances over rough ground, lifting/moving heavy weights, sitting for prolonged periods, walking long distances, frequent jumping between different levels, and standing for prolonged periods. Odds ratios (ORs) in the higher exposure categories ranged from 1.46 to 2.65. Cumulative exposure to three sporting activities was significantly associated with hip pain: track and field sports, jogging, and walking, with odds ratios varying between 1.57 to 1.94. On multivariate analysis three factors were independent predictors of hip pain onset: cumulative exposure of sitting for prolonged periods (higher exposure v not exposed: OR=1.82, 95% confidence interval (CI) 1.13 to 2.92), lifting weights >50 lb (23 kg) (OR=1.74, 95% CI 1.06 to 2.86) (both relating to the workplace), and walking as a leisure activity (OR=1.97, 95% CI 1.32 to 2.94). The population attributable risk associated with each of these activities was 21%, 13%, and 16%, respectively CONCLUSIONS: Cumulative exposure to some workplace and sporting "mechanical" risk factors for hip osteoarthritis (OA) appear to be related to hip pain in general-some (but not all) have previously been related to hip OA. Because these are common workplace or leisure time activities their attributable risk is high. PMID- 12634231 TI - Prospective comparative study of patients with culture proven and high suspicion of adult onset septic arthritis. AB - OBJECTIVE: To investigate whether patients with acute septic arthritis (SA) diagnosed by positive synovial fluid (SF) culture (Newman grade A) have different clinical and serological features from those with sterile SF in whom there is nonetheless a high suspicion of SA (Newman grades B and C). PATIENTS AND METHODS: A prospective 12 month multicentre hospital based study of adult patients with SA recruited 47 patients with culture positive SA and 35 patients with clinically suspected SA but sterile SF. RESULTS: Patient demography, clinical and laboratory features at presentation were similar irrespective of the underlying diagnosis, SF culture, and the presence of prosthetic joints. Medical and surgical treatment and outcome were comparable in the two patient groups. Patients with both suspected and proven SA were more likely to be from the more socially deprived areas of our community (p<0.0001). CONCLUSION: Patients in whom there is a high clinical suspicion of SA are comparable to those patients with SA with a positive SF culture and have similar morbidity and mortality on follow up. Therefore, if clinical suspicion of SA is high then it is correct to treat as SA in the absence of bacterial proof. PMID- 12634232 TI - Cartilage turnover assessed with a newly developed assay measuring collagen type II degradation products: influence of age, sex, menopause, hormone replacement therapy, and body mass index. AB - BACKGROUND: Cartilage normally has a slow turnover but in arthritis increased metabolism results in degradation of the tissue. OBJECTIVE: To assess cartilage turnover in a sample of the general population by an assay measuring cartilage derived urinary collagen type II (CTX-II) C-telopeptide degradation products. METHODS: CTX-II concentrations were measured in urine samples from 615 healthy men and women aged 20-87 years, and the influence of age, sex, menopause, hormone replacement therapy (HRT), and body mass index (BMI) was assessed. RESULTS: CTX II concentrations showed age dependent variations, with notable differences between men and women. Mean (SD) CTX-II concentration in postmenopausal women (220 (118) ng/mmol, n=25) was significantly higher than in an age matched group of premenopausal women (112 (79) ng/mmol, n=26, p<0.001). CTX-II concentration in women using HRT (118 (57) ng/mmol, n=50) was significantly lower than in an age and BMI matched group of women not receiving HRT (215 (99) ng/mmol, n=50, p<0.001). In subjects with a BMI >or=25 kg/m(2), CTX-II concentrations were significantly higher than in those with a BMI <25 kg/m(2) (185 (114) v 148 (91) ng/mmol, p<0.001). CONCLUSIONS: Cartilage turnover, as assessed by measuring urinary degradation products of CTX-II varies considerably with age, and significant differences between CTX-II levels in men and women as well as in pre- and postmenopausal women are found. Further studies are required to validate the marker for assessing cartilage degradation in arthritis. PMID- 12634233 TI - Can procalcitonin measurement help in differentiating between bacterial infection and other kinds of inflammatory processes? AB - OBJECTIVE: To study the levels of procalcitonin (PCT) in various inflammatory states seen in an internal medicine department and to evaluate the possible discriminative role of PCT in differentiating bacterial infection from other inflammatory processes. METHODS: PCT, C reactive protein (CRP), and white blood cell count (WBC) were measured in patients admitted to the department for fever or biological inflammatory syndrome, or both. The serum of 173 consecutive patients was analysed according to the aetiological diagnosis. The patients were divided into two groups: group I (n=60) with documented bacterial or fungal infection; group II (n=113) with abacterial inflammatory disease. RESULTS: PCT levels were >0.5 ng/ml in 39/60 (65%) patients in group I. In group II, three patients with a viral infection had slightly increased PCT levels (0.7, 0.8, and 1.1 ng/ml) as did two others, one with crystal arthritis and the other with vasculitis (0.7 ng/ml in both cases). All other patients in group II had PCT levels <0.5 ng/ml. In this study a value of PCT >0.5 ng/ml was taken as the marker of bacterial infection (sensitivity 65%, specificity 96%). PCT values were more discriminative than WBC and CRP in distinguishing a bacterial infection from another inflammatory process. CONCLUSION: PCT levels only rose significantly during bacterial infections. In this study PCT levels >1.2 ng/ml were always evidence of bacterial infection and the cue for starting antibiotic treatment. PMID- 12634235 TI - Increase in bone mineral density of patients with spondyloarthropathy treated with anti-tumour necrosis factor alpha. AB - OBJECTIVE: To determine the changes in bone mineral density (BMD) in patients with spondyloarthropathy (SpA) treated with infliximab. PATIENTS AND METHODS: 29 patients (six women; 23 men) aged 22-68 years, with persistently active SpA despite a high dose of non-steroidal anti-inflammatory drug and/or treatment with methotrexate or sulfasalazine, were studied. Median duration of disease was 13 years (range 3-30). Twenty five patients were treated with 5 mg/kg and four with 3 mg/kg of infliximab at weeks 0, 2, 6 and then received either no infusion (n=3), or additional infusion of infliximab every other month (n=6), and the remainder received one infusion only in the case of a relapse. Lumbar and femoral BMD was measured by dual energy x ray absorptiometry at baseline and six months later. Serum osteocalcin and urinary deoxypyridinoline were measured in 19 patients at weeks 0, 2, 24, and in 13 patients at all visits. RESULTS: In six months there was a significant increase in BMD at the spine (3.6%, p=0.001), total hip (2.2%, p=0.0012), and trochanter (2.3%, p=0.0012). A trend for increase (1.1%) was observed at the femoral neck. There was an increase in osteocalcin between baseline and week 6 (third infusion)-median 1.45 micro g/l (p=0.013). No change in marker of bone resorption was observed at the same time. There was no change in biochemical markers between baseline and final visits. There was a trend for a correlation between the decrease at six months in erythrocyte sedimentation rate, and lumbar spine BMD change (r(s)=-0.35, p=0.06). CONCLUSION: These data suggest that a benefit of anti-tumour necrosis factor alpha therapy on BMD in patients with SpA may be through an uncoupling effect on bone cells. PMID- 12634234 TI - Beneficial effects of the anti-oestrogen tamoxifen on systemic lupus erythematosus of (NZBxNZW)F1 female mice are associated with specific reduction of IgG3 autoantibodies. AB - BACKGROUND: Sex hormones have been shown to influence the immune system and to modify the course of autoimmune disorders. OBJECTIVE: To examine the effects of the oestrogen antagonist tamoxifen on the course of systemic lupus erythematosus (SLE) in (NZBxNZW)F1 mice. METHODS: Groups of 8 week old (NZBxNZW)F1 female mice were treated with tamoxifen (800 micro g/mouse; twice a week) or with double distilled water for four months. Mice were evaluated monthly for the presence of autoantibodies directed against DNA and nuclear extract (NE) by enzyme linked immunosorbent assay (ELISA). White blood cells and thrombocytes were quantified by a cell counter and proteinuria by combistix kit. At 6 months of age, all mice that did not die spontaneously were killed and evaluated for the presence of glomerular immune deposits by indirect immunofluorescence assay. IgG isotypes of autoantibodies in the mouse sera and glomeruli were determined by gamma chain specific antibodies. RESULTS: Tamoxifen treatment significantly reduced autoantibody production directed against either NE or DNA. The latter reduction was mainly in autoantibodies of the IgG3 isotype. Furthermore, tamoxifen had significant beneficial effects on the course of SLE in (NZBxNZW)F1 mice. At 6 months of age, 40% of the untreated mice died spontaneously, whereas all the tamoxifen treated mice were still alive. All untreated mice showed severe thrombocytopenia and persistent proteinuria, with diffuse glomerular immune deposits of IgG2a and IgG3 isotypes in their kidneys. In contrast, the tamoxifen treated mice had a normal number of thrombocytes and only minimal proteinuria. Moreover, glomerular immune deposits were detected in <40% of the tamoxifen treated mice. The latter were mainly of the IgG2a but not of the IgG3 isotype. CONCLUSION: The results clearly show the remarkable therapeutic effects of tamoxifen on SLE of (NZBxNZW)F1 female mice and suggest that these beneficial effects are related to the specific reduction of IgG3 autoantibodies. PMID- 12634236 TI - Oral corticosteroid prescribing in women over 50, use of fracture prevention therapy, and bone densitometry service. AB - OBJECTIVE: To identify the most common diseases and age of corticosteroid use in women over 50, dosage in last year, duration of oral corticosteroid use, prescription for fracture prevention (drug used), and referrals for bone densitometry. METHODS: General practice records from 41 practices in Shropshire identified 62,230 women aged >50 from a population of 80,082. Data on fractures, duration of corticosteroid use, dose in the study year (1 April 1997-31 March 1998), use of fracture prevention therapy and bone densitometry were sampled from one out of three records. RESULTS: 3.2% were prescribed corticosteroids; 633 patients investigated in detail aged 70.1 (SD 10.5) years, had been prescribed 1526 (SD 1727) mg prednisolone (median 1040 mg) for 3.31 (SD 3.20) years (median 2.0 years). Patients with asthma/lung disease, most common in the younger group, had the lowest annual corticosteroid use; patients with rheumatoid arthritis (RA), polymyalgia rheumatica/temporal arteritis (PMR/TA), who were more likely to be elderly, had the highest annual use. Between the age of 70 and 79 years patients with RA had significantly more hip fractures than the other groups, and corticosteroid prescribing was most common. Bisphosphonates or hormone replacement therapy were prescribed for 48% aged 50-59 years but only 32% at 70 79 years (p<0.01); patients with asthma and RA being less likely recipients (p<0.01). Referrals for bone densitometry had occurred in 20.2%,with 60.2% having osteoporosis. Referrals were more common in those taking corticosteroids for longer periods (p<0.01). CONCLUSIONS: The elderly had the most prescriptions for corticosteroid treatment but the fewest for effective fracture prevention therapy. Patients with RA, PMR/TA had the greatest corticosteroid dosage, for the longest time. Patients with RA sustained more hip fractures than other groups but were least likely to have effective fracture prevention therapy prescribed. PMID- 12634237 TI - Regional differences in Finland in the prevalence of rheumatoid factor in the presence and absence of arthritis. AB - OBJECTIVES: To look for possible regional differences in the prevalence of rheumatoid factor (RF) in the presence and absence of arthritis. METHODS: The study covered a representative sample of the Finnish population aged 30 years or over, primarily comprising 8000 people, of whom 7217 participated in the field survey carried out in 1978-80. RF from serum samples from 7116 subjects was determined by the Waaler-Rose (sensitised sheep cell agglutination) test. Titres >or=32 were regarded as positive and titres >or=128 as strongly positive. Arthritis was diagnosed by a thorough clinical examination. RESULTS: In the absence of arthritis the prevalence of positive and strongly positive RF reactions was 2.1% and 1.0%, respectively. The lowest prevalence of strongly "false positive" RF occurred in south western Finland. After adjustment for age, sex, smoking, and coffee consumption the odds ratio of having a strongly "false positive" RF reaction in eastern Finland was 3.16 (95% confidence interval 1.29 to 7.72) and in northern Finland 2.94 (1.13 to 7.64) compared with south western Finland. The corresponding odds ratio of strongly RF positive arthritis in eastern Finland was 5.08 (1.41 to 18.27). CONCLUSION: Regional differences are found in the prevalence of a strongly positive RF reaction in the Finnish population. The findings are in accordance with recent results from another study concerning regional differences in the incidence of rheumatoid arthritis in Finland. PMID- 12634238 TI - Syndrome of symptomatic adult acetabular dysplasia (SAAD syndrome). AB - OBJECTIVES: To determine the prevalence of acetabular dysplasia in subjects presenting with hip pain to primary care and its relationship with radiographic osteoarthritis (OA) of the hip. DESIGN: Cross sectional analysis of a prospective cohort. SETTING: 35 general practices across the UK. SUBJECTS: 195 patients (63 male, 132 female) aged 40 years and over presenting with a new episode of hip pain RESULTS: The prevalence of acetabular dysplasia in this study of new presenters with hip pain was high (32%). There was no significant relationship between acetabular dysplasia and radiographic OA overall. CONCLUSIONS: The high prevalence of acetabular dysplasia across all grades of OA severity suggests that dysplasia itself may be an important cause of hip pain ("symptomatic adult acetabular dysplasia"). PMID- 12634239 TI - Interleukin 1beta and tumour necrosis factor alpha secreting cells are increased in the peripheral blood of patients with primary Sjogren's syndrome. AB - OBJECTIVE: To study systemic alterations of cytokine secreting peripheral blood mononuclear cells (PBMC) in primary Sjogren's syndrome (pSS) and their relation to common clinical and immunological manifestations of this disease. METHODS: PBMC spontaneously secreting tumour necrosis factor alpha (TNFalpha), interleukin 1beta (IL1beta), and interleukin 6 (IL6) were assessed by enzyme linked immunospot (ELISPOT) analysis in a cohort of 31 patients with pSS fulfilling the modified European classification criteria. Nineteen healthy volunteers served as controls. ELISPOT results were correlated with glandular and extraglandular manifestations and autoantibody titres-that is, rheumatoid factor (RF) isotypes, anti-Ro/SS-A, anti-La/SS-B as determined by an enzyme linked immunosorbent assay (ELISA) technique. RESULTS: The number of TNFalpha and IL1beta secreting cells was significantly higher in patients with pSS than in controls. No differences were detected in the number of IL6 secreting PBMC. Patients with recurrent parotid swelling (RPS) had a significantly increased number of IL1beta secreting PBMC. Moreover, the number of IL1beta secreting PBMC correlated with the disease duration (r(s)=0.479; p<0.01) and with the concentration of IgM RF (r(s)=0.63; p<0.01) and IgG RF (r(s)=0.42; p<0.05). Other autoantibodies did not correlate with cytokine secreting PBMC. CONCLUSION: The increased systemic secretion of IL1beta and TNFalpha in patients with pSS points to a pathogenic impact of these cytokines in this autoimmune disease. In particular the correlation of IL1beta secreting PBMC with RPS and RF production indicates that IL1beta is a crucial regulator in the development of local and systemic disease manifestations. PMID- 12634240 TI - TAP1 and TAP2 polymorphisms analysis in northwestern Colombian patients with systemic lupus erythematosus. AB - OBJECTIVE: To determine the influence of TAP1 and TAP2 alleles in northwestern Colombian patients with systemic lupus erythematosus (SLE). METHODS: Unselected patients with SLE (n=140) and controls (n=120) matched for sex, age, and ethnicity were analysed. Clinical manifestations, clinical activity, and severity of disease were recorded. Autoantibodies were detected by enzyme linked immunosorbent assay (ELISA). TAP1 and TAP2 polymorphisms were determined by amplification refractory mutation system-polymerase chain reaction. A Hardy Weinberg equilibrium test, microdifferentiation analysis, linkage disequilibrium analysis, and haplotype and allele frequency comparisons were performed. RESULTS: The TAP2 variant Val379/Ala565/Ala665 (allele TAP2*0201) was associated with SLE (56% v 39%; odds ratio=2, 95% confidence interval 1.22 to 3.30, p(c)=0.03). There was no stratification between patient and control samples. Linkage disequilibrium between TAP1 and TAP2 loci was found in controls but not in patients. An excess in the number of heterozygotes in the TAP2 locus was found in patients. No association between TAP1 and TAP2 variants and the presence of autoantibodies, clinical expression, or severity of disease was found. CONCLUSIONS: The TAP2 locus influences susceptibility to SLE in our patient group; however, it has no significant effect on the immune response or on the clinical course of the disease. PMID- 12634241 TI - Abrupt condylar destruction of the mandibula in juvenile idiopathic arthritis. PMID- 12634242 TI - Benefit of anti-TNFalpha treatment for nephrotic syndrome in a patient with juvenile inflammatory bowel disease associated spondyloarthropathy complicated with amyloidosis and glomerulonephritis. PMID- 12634243 TI - A unique autoantibody pattern of positive anti-Jo-1, anti-U1RNP, and antiproteasome antibodies in autoimmune myositis as a diagnostic challenge. PMID- 12634244 TI - Degenerative disc disease and pre-existing spinal pain. PMID- 12634245 TI - Tuberculous tonsillitis in a patient receiving etanercept treatment. PMID- 12634246 TI - Successful treatment of resistant giant cell arteritis with etanercept. PMID- 12634247 TI - Bilateral subdural effusion in a patient with neuro-Behcet's disease. PMID- 12634248 TI - A case of multiple sclerosis associated with rheumatoid arthritis and positive anticardiolipin antibodies. PMID- 12634249 TI - Observation on serum anti-double stranded DNA antibodies of tripterine in systemic lupus erythematosus of (NZBxW)F1 mice. PMID- 12634250 TI - Clodronate induced uveitis. PMID- 12634251 TI - Aminotransferase levels during treatment of rheumatoid arthritis with leflunomide in clinical practice. PMID- 12634252 TI - Magnetic resonance imaging of the hand in mixed connective tissue disease. PMID- 12634253 TI - Power Doppler sonography: synovial tissue assessment in RA. PMID- 12634254 TI - A case of orbital myositis associated with rheumatoid arthritis. PMID- 12634255 TI - Murine retrovirus infection and the effect of chronic alcohol consumption: proteomic analysis of cardiac protein expression. AB - AIMS: The cardiovascular complications of acquired immunodeficiency syndrome (AIDS) are serious, including the occurrence of pathological heart conditions such as cardiomyopathy. Chronic alcohol consumption accentuates the severity of AIDS and may contribute to the development of cardiomyopathy. The aim of this work was to use a proteomics approach to investigate global alterations in protein expression in a mouse model of AIDS in the presence or absence of chronic alcohol consumption. METHODS: Cardiac proteins were separated by two-dimensional polyacrylamide gel electrophoresis and quantitative computer analysis was used to evaluate the resulting two-dimensional protein profiles. Proteins that were differentially expressed in the hearts of mice from the different experimental groups were identified by peptide mass finger-printing by matrix-assisted laser desorption/ionization mass spectrometry. RESULTS: A number of specific proteins were observed to be differentially expressed in the mouse heart due to the effect of ethanol feeding alone. Differentially expressed proteins were also observed that were due to viral infection alone. Ethanol feeding and viral infection appeared to have similar effects on the expression of a number of proteins. A total of 24 proteins were altered by infection alone. Of these 24 proteins, eight were affected by alcohol, with six alterations being ameliorated and two being exacerbated by alcohol. Two of these proteins have been identified as the 27 kDa heat-shock protein and mitochondrial long-chain acyl-CoA thioesterase 1. CONCLUSIONS: These results suggest that chronic alcohol consumption may exacerbate the effects of viral infection on the heart by lowering the stress response leading to de-protection and further cytotoxic effects. PMID- 12634256 TI - Neutrophil activation by murine retroviral infection during chronic ethanol consumption. AB - AIMS: Neutrophil adhesion molecule CD11b and reactive oxygen species (ROS) are neutrophil activation markers for evaluating the functional activity of neutrophils. The aim of this study was to determine if neutrophils are activated in murine AIDS and/or chronic ethanol consumption and if neutrophil CD11b expression and ROS production vary when progressive retrovirus infection occurs. METHODS: Four groups were studied: control, murine AIDS, ethanol and ethanol plus murine AIDS. Neutrophil activation was assessed by CD11b expression and ROS production using flow cytometry. RESULTS: We found that neutrophils lost their responsiveness to fMLP due to retrovirus or ethanol exposure. In the murine AIDS group, neutrophil CD11b expression was up-regulated along with a significant increase in ROS after 1 month of retroviral infection. After 2 months, neutrophil CD11b and ROS decreased. However, neutrophil CD11b expression further increased after 3 months. In the ethanol consumption group, neutrophil CD11b expression was down-regulated after 2 months, whereas ROS production increased in the first and third months. In the murine AIDS plus ethanol group, there were significant increases in both ROS and CD11b expression during the 3-month observation period. CONCLUSIONS: These findings suggest that neutrophil function is impaired by LP BM5 retrovirus infection and/or chronic ethanol consumption. The pattern of neutrophil CD11b expression and ROS production might help to predict the stage of murine AIDS. Ethanol may further compromise neutrophil function in AIDS. PMID- 12634257 TI - Continuous ethanol administration influences rat brain 5-hyroxytrytamine synthesis non-umiformly: alpha-[14C]methyl-L-trytophan autoradiographic measurements. AB - AIMS: The influence of alcohol on the brain serotonergic system has been studied for several decades with some discordant results. The effects of continuous and constant treatment with ethanol on the rates of serotonin [5-hydroxytryptamine (5 HT)] synthesis in discrete regions of the rat brain were studied. METHODS: 5-HT synthesis rates were measured using the alpha-[(14)C]methyl-l-tryptophan autoradiographic method. The rats in the experimental group were treated with 50% ethyl alcohol and those in the control group received distilled water. The fluid was delivered subcutaneously by implanted osmotic mini-pumps for 14 days at the rate of 5 micro l/h or 0.12 ml/day (0.06 ml of alcohol per day). RESULTS: Chronic ethanol treatment, as delivered in the present experiment, induced a significant increase in the rate of 5-HT synthesis in descending serotonergic cell bodies (raphe pallidum, raphe obscurus, raphe magnus), nigrostriatal structures, the hippocampus and cortices. No significant changes were observed in the dorsal and median raphe nuclei or pineal body. The results suggest that there may be differences in the regulation of 5-HT synthesis in different brain structures after 14 days of continuous (subcutaneous) injection of 50% alcohol. CONCLUSIONS: Chronic ethanol treatments using osmotic mini-pumps induce non-uniform increases in 5-HT synthesis in the rat brain. PMID- 12634258 TI - Joint use of clinical parameters, biological markers and CAGE questionnaire for the identification of heavy drinkers in a large population-based sample. AB - AIMS: Alcohol consumption in France is one of the highest in the world. Factors associated with excessive alcohol drinking are numerous. However, taken separately, none of the existing clinical or biological markers of excessive alcohol intake enables an adequate identification of heavy drinkers. The aim of this cross-sectional survey was to identify socio-demographic, clinical and biological factors associated with excessive alcohol drinking, to develop a model and to assess its reliability, thus enabling the detection of heavy drinkers. METHODS: Subjects were 1619 men and 1559 women, aged 35-64 years, living in three French areas (Lille, Strasbourg and Toulouse) and randomly selected from polling lists. Socio-demographic status, lifestyle, reported alcohol intake and answers to the CAGE questionnaire (alcohol dependence) were obtained by questionnaire. A blood sample was taken for quantification of biological parameters. Men who drank 60 g of ethanol a day (g/day) or above and women who drank 30 g/day or above were classified as heavy drinkers. The reference class (RC) gathered non-drinkers and moderate drinkers together. The sample was divided into two sub-samples: the first was used to estimate the parameters of a logistic regression model (heavy drinkers vs others), and the second to assess the accuracy of this model for the identification of heavy drinkers, using receiver operating characteristic (ROC) curves. A specific analysis was performed for each gender. RESULTS: Fourteen per cent of men and 40.8% of women were non-drinkers. Nine per cent of women and 14.4% of men were heavy drinkers. Wine was the most consumed alcoholic beverage. In the univariate analyses, differences were observed between the two groups of alcohol consumers for most of the socio-demographic, clinical and biological variables considered. In the multivariate analyses, low educational level, smoking, apoprotein B, high density lipoprotein cholesterol, mean corpuscular volume (MCV), gamma-glutamyl-transferase (GGT) and the CAGE score for men, and living area, age, MCV, GGT and the CAGE score for women remained independently and significantly associated with heavy drinking. In the validation sub-sample, these models combining different types of markers enabled a good discrimination between heavy drinkers and the RC, with an area under the ROC curve of 82% for men and of 79% for women. CONCLUSIONS: In this study, socio-demographic, clinical and biological factors and the CAGE score were independently related to excessive alcohol drinking and their joint utilization in a screening model enabled a good recognition of heavy drinkers. PMID- 12634259 TI - Estimating non-response bias in a survey on alcohol consumption: comparison of response waves. AB - AIMS: According to 'the continuum of resistance model' late respondents can be used as a proxy for non-respondents in estimating non-response bias. In the present study, the validity of this model was explored and tested in three surveys on alcohol consumption. METHODS: The three studies collected their data by means of mailed questionnaires on alcohol consumption whereby two studies also performed a non-response follow-up. RESULTS: Comparisons of early respondents, late respondents and non-respondents in one study showed some support for 'the continuum of resistance model', although another study could not confirm this result. Comparison of alcohol consumption between three time response groups showed no significant linear pattern of differences between response waves. CONCLUSIONS: The hypothesis that late respondents are more similar to non respondents than early respondents, could not be confirmed or rejected. Repeated mailings are effective in obtaining a greater sample size, but seem ineffective in improving the representativeness of alcohol consumption surveys. PMID- 12634260 TI - Acamprosate in Korean alcohol-dependent patients: a multi-centre, randomized, double-blind, placebo-controlled study. AB - AIMS: A multi-centre, randomized, double-blind, placebo-controlled trial was conducted to evaluate the efficacy and the safety of acamprosate over 8 weeks in Korean alcohol-dependent patients. METHODS: One hundred and forty-two alcohol dependent patients in 12 centres were randomized to 8 weeks treatment with either acamprosate (n = 72) or a placebo (n = 70) in combination with out-patient psychosocial intervention. They were predominantly male (95.8%), with a mean age of 44.3 +/- 8.3 years; 76.1% were married; 59.9% were employed; 58.5% had received previous alcoholism treatment (previous mean number of admissions in alcoholism in-patient programmes 4.6 +/- 6.9). At visits to the clinic (weekly for 4 weeks, then biweekly for 4 weeks), a record was made of alcohol use (Time Line Follow-Back), alcohol craving using a Korean version of the Obsessive Compulsive Drinking Scale and a visual analogue scale, and adverse events. Serum aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltransferase (GGT), blood urea nitrogen and creatinine levels were measured on weeks 0, 2, 4 and 8. RESULTS: In the acamprosate group (A), 71.4% had had alcohol within the 2 days prior to starting medication, against 65.2% of patients in the placebo group (P); (P > 0.05). One hundred and one subjects (71.1%) completed 8-weeks of treatment (A, 73.6%; P, 68.6%; P > 0.05). During the 8-week treatment period, 37, (A) (n = 72) and 32% (P) (n = 70) achieved continuous abstinence (P > 0.05), and 40, (A) and 39% (P) remained without relapse (P > 0.05) (defined as a day when a man consumed five or more drinks or a woman four or more drinks). The percentage of days abstinent during the 8-week treatment period was 81.2, (A) and 78.5% (P) (P > 0.05), and the percentage of days without heavy drinking 86.1 (A) and 84.9% (P) (P > 0.05). The mean amount drunk per drinking occasion was 7.2, (A) and 8.6 standard drinks (P) (P > 0.05). No statistically significant differences in changes in the serum GGT level or craving scores from baseline to the end-point of treatment were found between the two groups. Recency of drinking prior to commencing study drug predicted percentage of days abstinent in the first 2 weeks on treatment; however, when ANOVAs were conducted using treatment outcomes as a dependent variable, medication condition as an independent variable and the period of abstinence prior to treatment as a covariate, a significant effect of medication condition was still not seen. CONCLUSIONS: Acamprosate was ineffective in reducing drinking in this Korean sample. The result differs from that of most European acamprosate trials. This might be explained by our sample's relatively severe alcohol dependence, and low social support, or the fact that many patients were still drinking near to their first medication. The variability of the psychosocial support, ethnicity (which might also affect acamprosate pharmacokinetics) and the Korean drinking style, which differs from that of Europeans, might have contributed to our negative result. PMID- 12634261 TI - An analysis by birth cohort of alcohol consumption by adults in Great Britain 1978-1998. AB - AIMS AND METHODS: The General Household Survey for Great Britain has, since 1978, in even numbered years, included questions on alcohol consumption. A cohort analysis was applied to these sequential cross-sectional data. RESULTS AND CONCLUSIONS: Analysis confirms that, after about age 20 years, both men and women, as they grow older, become more likely to be non- or very light drinkers. Men are also less likely to be heavy drinkers as they grow older, but for women the picture is less clear-cut. Cohort effects can also be discerned, but are much less obvious than for smoking. There is a general trend for members of later cohorts at any age to be less likely to be non- or very light drinkers and more likely to be heavy drinkers, than members of earlier cohorts. However, for men, this trend seems to have ended with those born from about 1927 to 1931, while for women the trend has continued and possibly become stronger in cohorts born after 1927-1931. PMID- 12634262 TI - Ethnic differences in cirrhosis of the liver in a British city: alcoholic cirrhosis in South Asian men. AB - AIMS AND METHODS: We studied the ethnic origin of cirrhotic patients retrospectively over the 14-year period 1987-2000 and compared the ethnic make-up of the cirrhotic patients with the ethnic make-up of the local catchment population. RESULTS AND CONCLUSIONS: Of 381 cirrhotics, 64.1% were white, 29.1% South Asian, 4.7% Afro-Caribbeans and 2.1% other races. These proportions were different from those of the local community in that South Asians were over represented and Afro-Caribbeans were under-represented. Alcohol was the commonest cause of cirrhosis (60.9%) and South Asian non-Moslem males with alcoholic cirrhosis were over-represented and were younger at diagnosis than white alcoholic cirrhotics. PMID- 12634263 TI - Alcohol intoxication reduces impulsivity in the delay-discounting paradigm. AB - AIMS: To examine the moderating effects of alcohol myopia on cognitive impulsivity in humans using the delay-discounting paradigm. METHODS: Seventy-six male undergraduate students were randomly assigned to sober, placebo or alcohol conditions. In the delay-discounting task, participants made a series of hypothetical choices between a small, immediate reward and a large, delayed reward. To test the predictions of alcohol myopia theory, participants completed a standard version of the task or one containing cues which impelled the impulsive choice (i.e. preference for the small, immediate reward). Participants also completed a personality measure of impulsivity and the go/no-go task, which assesses motor impulsivity. RESULTS: Intoxicated participants tended to discount delayed rewards at lower rates than sober participants, and blood alcohol level was inversely correlated with delay discounting. The impelling cues did not moderate the effects of alcohol on delay discounting. CONCLUSIONS: Alcohol intoxication does not always increase cognitive impulsivity and may lead to more cautious decision-making under certain conditions. PMID- 12634264 TI - Lipoprotein(a) in alcohol-dependent male patients during a six-month abstinence period. AB - AIMS: The best known and probably most important mechanism of health-protective moderate alcohol drinking is beneficial changes in plasma lipid levels. We determined changes in main plasma lipid levels in alcohol-dependent patients over a 6-month abstinence period. METHODS: Fifty-four alcohol-dependent male patients, who were abstinent for less than 14 days, and 20 non-alcoholic males, who had not drunk alcohol for the last month, were studied. In all patients at the study start and after 4 weeks and 6 months observation, lipoprotein(a) [Lp(a)], total cholesterol (TC), HDL cholesterol (HDL), LDL cholesterol (LDL) and triglyceride concentrations, both fasting and 5 h after a fatty meal, were determined. RESULTS: Alcohol-dependent patients had similar mean fasting and post-prandial plasma lipid levels as the control group, both at the study start and after 4 weeks of abstinence. Whereas, in alcohol-dependent patients after 4 weeks of abstinence, a significant decrease in Lp(a) and fasting HDL levels, as well as a significant increase in fasting LDL level and pro-atherogenic indices of plasma lipids [TC/HDL, (TC-HDL)/HDL, LDL/HDL, Lp(a)/HDL] were observed. Post-prandial levels of studied plasma lipids, except HDL, did not change over the 6-month observation period. In patients who did not remain abstinent for the whole observation period (n = 9), in comparison to abstinent patients, significantly higher HDL levels and a tendency to higher values of LDL, LDL/HDL, Lp(a) and Lp(a)/HDL were found. CONCLUSIONS: (1) Higher Lp(a) levels soon after alcohol withdrawal may be a factor potentially responsible for the increase of acute cardiac syndromes prevalent in the drinking and early abstinence period, in spite of high HDL concentration; (2) in alcohol-dependent male patients, after a 6 month abstinence period, pro-atherogenic plasma cholesterol fraction changes occurred, expressed by a decrease in HDL level and an increase in LDL concentration. PMID- 12634265 TI - Fatty Acid ethyl esters in scalp, pubic, axillary, beard and body hair as markers for alcohol misuse. AB - AIMS: This study examined the possibility of using hair samples other than scalp hair for analysis of fatty acid ethyl esters (FAEE) as markers for alcohol misuse. METHODS: Samples of scalp and pubic hair and, if available, axillary, beard and body hair of one teetotaller, five moderate social drinkers and 22 fatalities were analysed for FAEE by head-space solid-phase microextraction and gas chromatography-mass spectrometry. The sum of the concentrations of ethyl myristate, ethyl palmitate, ethyl oleate and ethyl stearate (C(FAEE)) in the hair samples was compared with information about the drinking behaviour of the individuals. RESULTS: Although there were large differences in the analyte concentrations in hair from different sites in the same individual, cases of chronic excessive alcohol consumption were characterized by C(FAEE) > 1.0 ng/mg in almost all samples. CONCLUSION: FAEE concentrations in hair other than scalp hair can be used as markers of chronic alcohol misuse. PMID- 12634267 TI - Impact of an alcohol misuse intervention for health care workers--1: Frequency of binge drinking and desire to reduce alcohol use. AB - AIMS: This report presents results of a 3-year study to evaluate the effects of an enhanced substance misuse prevention/early intervention programme on binge drinking and desire to reduce alcohol use among health care professionals employed in a managed care organization. METHODS: The intervention was implemented at one site, but not at satellite locations, which were used for comparison. The intervention included relatively low-cost elements, such as substance misuse awareness training for managers and the use of health risk appraisals (HRAs) and educational videos on how to reduce stress, depression and binge drinking. We evaluated intervention effects by comparing HRA scores of employees at the intervention site to all other employees who completed the HRA, while adjusting for demographic factors, reported stress levels, employment site and the effects of time. Outcomes analysed included number of days binge drinking (drinking 5 or more drinks per occasion in the past 30 days) and desire to reduce alcohol use. RESULTS: Binge drinking rates were not affected by the intervention. Among those who binge-drank, however, employees who completed the HRA at the intervention site in the post-intervention period were 2.59 times more likely to report a desire to cut down on alcohol use, compared with the pre-intervention time period and with both time periods in the comparison site (P < 0.05). CONCLUSIONS: We conclude that while the intervention did not significantly affect reported alcohol use, it did increase motivation to reduce alcohol use among binge drinkers. PMID- 12634266 TI - Alcohol withdrawal treatment in intoxicated vs non-intoxicated patients: a controlled open-label study with tiapride/carbamazepine, clomethiazole and diazepam. AB - AIMS AND METHODS: Alcohol withdrawal treatment efficacy of tiapride/carbamazepine (A) vs clomethiazole (B) vs diazepam (C) in non-intoxicated patients and vs tiapride/carbamazepine in intoxicated patients (D; breath alcohol concentration > or = 1 g/l) was tested (n = 127) in a controlled randomized open-label study. RESULTS: Efficacy and safety were not different between groups (total group: delirium, 3.9%; seizure, 0.8%), except for a lack of efficacy in 18% of intoxicated tiapride/carbamazepine patients. A change of medication in this group was necessary only when primarily intoxicated patients had reached the non intoxicated range. CONCLUSIONS: Treatment with tiapride/carbamazepine in alcohol intoxicated patients proved to be safe. PMID- 12634268 TI - Impact of an alcohol misuse intervention for health care workers --2: Employee assistance programme utilization, on-the-job injuries, job loss and health services utilization. AB - AIMS: We evaluated the effects of an enhanced substance misuse (SM) prevention/early intervention programme on referrals to an employee assistance programme, health care utilization rates, on-the-job injury rates and job termination rates among health care professionals employed in a managed care organization. METHODS: The intervention was implemented at one site, with the remaining sites serving as the comparison group. Existing data from hospital databases were used to compare events occurring in the periods before and after initiation of the intervention. To account for baseline differences in age, gender and job class, logistic regression models produced adjusted means for events per employee month-at-risk. RESULTS: We found that employee assistance referrals and non-SM-related in-patient hospitalizations increased significantly post-intervention, while rates of total out-patient SM-related visits decreased at both the intervention and comparison sites post-intervention. There was a small, statistically significant decrease in the monthly rate (OR = 0.92) of non SM out-patient utilization at the intervention site, once the intervention was in place. No differences potentially attributable to the intervention were detected in job turnover or injury rates. CONCLUSIONS: We conclude that, while the intervention did not appear to affect health care utilization for SM-related problems, it was associated with increased referrals for employee assistance. PMID- 12634269 TI - Impaired serum cortisol stress response is a predictor of early relapse. AB - AIMS: to investigate a possible association of cortisol stress response during early abstention with relapse. METHODS: Thirty-six alcohol-dependent males, half of them with a comorbid anxiety disorder, and 15 healthy controls were exposed to a standardized psychosocial stress test. Thirty-one of the patients were assessed for relapse 6 weeks after discharge. RESULTS: The relapsers showed almost no cortisol responses in the stress test. Comorbid anxiety disorder influenced neither stress response nor relapse. CONCLUSIONS: During early abstention from alcohol, reduced stress-responsivity of the hypothalamo-pituitary-adrenocortical axis seems to be connected to early relapse. PMID- 12634271 TI - One hundred alcoholic doctors: a 21-year follow-up. PMID- 12634272 TI - Visceromotor and spinal neuronal responses to colorectal distension in rats with aldosterone onto the amygdala. AB - Stereotaxic delivery of corticosterone onto the amygdala produces colorectal hypersensitivity through activation of lumbosacral spinal neurons. Since corticosterone activates both the mineralocorticoid (MR) and glucocorticoid (GR) receptors, the aim of this study was to determine the importance of MRs in the regulation of colorectal hypersensitivity through the use of aldosterone that preferentially binds to MRs. Fischer-344 rats received either aldosterone (n = 18)- or cholesterol (control, n = 18)-containing micropellets bilaterally placed stereotaxically on the dorsal margin of the amygdala. After 1 wk, colorectal sensitivity to distension (30 mmHg) was measured in a subgroup of rats (n = 8/group). In other rats (n = 10/group), extracellular potentials of single L6-S1 spinal neurons in response to colorectal distension (CRD; 10-80 mmHg) were recorded. In aldosterone-implanted rats, CRD produced a greater visceromotor behavioral response compared with cholesterol controls (19 +/- 0.5 vs. 11.5 +/- 2.7; P < 0.01). A total of 68/182 (37%) and 56/167 (34%) of spinal neurons responded to noxious CRD in aldosterone-implanted and control groups, respectively. A total of 36/42 (86%) neurons excited by CRD had spontaneous activity in aldosterone-implanted groups compared with control (19/33, 58%, P < 0.01). Neurons with low thresholds for excitatory responses to CRD were seen more frequently in aldosterone-implanted rats than those in the control group (35/39 vs. 18/31, P < 0.05). Maximal excitatory responses of neurons to CRD in aldosterone-implanted rats were significantly greater (23.9 +/- 2.2 vs. 16.4 +/- 2.0 imp/s, P < 0.05), and the durations were longer (34.3 +/- 2.7 vs. 24.9 +/- 1.4 s, P < 0.05) than those in control group. Finally, a greater number of neurons had wide dynamic range responses to somatic stimulation in aldosterone treated rats compared with cholesterol controls. Our findings suggest that, in the amygdala, MR receptor-mediated mechanisms are likely involved in descending pathways onto lumbosacral spinal neurons that induce colorectal hypersensitivity to luminal distension. PMID- 12634273 TI - Cyclic h-reflex modulation in resting forearm related to contractions of foot movers, not to foot movement. AB - During rhythmic voluntary oscillations of the foot, the excitability of the H reflex in the Flexor Carpi Radialis (FCR) muscle of the resting prone forearm increases during the foot plantar-flexion and decreases during dorsiflexion. It is known that, when the two extremities are moved together, isodirectional (in phase) coupling is the preferred form of movement association. Thus the above pattern of the H-reflex excitability modulation may favor the preferred coupling between the two limbs. To gain some clues about its origin, FCR H-reflex excitability was tested before and after modifying the phase relations between the activation [electromyogram (EMG)] of foot movers and foot movement, either by loading of the foot or by changing the movement frequency. After foot loading, the movement cycle was consistently delayed with respect to the onset of the EMG in Soleus (Sol) or Tibialis Anterior (TA) muscles. Simultaneously, the FCR H reflex modulation advanced by that same amount with respect to the foot movement, thus remaining phase-locked to the EMG onsets. Similarly, when movement frequency was varied step-wise between 1.0 and 2.0 Hz, the foot movement was progressively delayed with respect to both the EMG onset (Sol and TA) and the FCR H-reflex modulation, so that the phase relation between the motor command to the foot and the H-modulation in the forearm remained constant. These results suggest that modulation of H-reflex in the forearm is tied to leg muscle contraction, rather than to foot kinematics, and point to a central, rather than kinesthetic, origin for the modulation. PMID- 12634274 TI - Visual-ocular motor activity in the macaque pregeniculate complex. AB - The anatomical connections of the pregeniculate complex (PrGC) with components of the visual-ocular motor system suggested its contribution to ocular motor behavior. Subsequent studies reported saccade-related activity in the primate PrGC. To determine its contribution, we characterized pregeniculate units (n = 128) in alert macaques during ocular motor tasks and visual stimulation. We found that 36/109 saccade-related units exhibited postsaccadic bursts or pauses in tonic discharge for saccades of any amplitude or direction. In contrast to previous results, 46/109 responses preceded or coincided with the saccade, while 47/109 responses were directionally tuned. Pregeniculate units were modulated not only in association with saccades (109/128) but also with smooth eye movements and visual motion (20/128) or eye position (23/128). Multiple ocular motor signals were recorded from 19% of the units, indicating signal convergence on individual neurons. Visual responses were demonstrated in 51% of PrGC units: visual field illumination modulated the resting discharge of 33 units; the responses of 37 saccade-related units and all 23 position-dependent units were modulated by visual stimulation. Early saccadic activity in the PrGC suggests that it contributes more to gaze than postsaccadic modulation of visual or ocular motor activity. The patterns of saccadic responses and the modulation of PrGC activity in association with a variety of visual-ocular motor behaviors suggest its potential role as a relay between the parietal cortex and elements of the brain stem ocular motor pathways, such as the superior colliculus and pretectal nucleus of the optic tract. PMID- 12634275 TI - Mechanism for activation of locomotor centers in the spinal cord by stimulation of the mesencephalic locomotor region. AB - The synaptic pathways of mesencephalic locomotor region (MLR)-evoked excitatory and inhibitory postsynaptic potentials (EPSPs and IPSPs) recorded from lumbar motoneurons of unanesthetized decerebrate cats during fictive locomotion were analyzed prior to, during, and after cold block of the medial reticular formation (MedRF) or the low thoracic ventral funiculus (VF). As others have shown, electrical stimulation of the MLR typically evoked short-latency excitatory or mixed excitatory/inhibitory PSPs in flexor and extensor motoneurons. The bulbospinal conduction velocities averaged approximately 88 m/s (range: 62-145 m/s) and segmental latencies for EPSPs ranged from 1.2 to 10.9 ms. The histogram of segmental latencies showed three peaks, suggesting di-, tri-, and polysynaptic linkages. Segmental latencies for IPSPs suggested trisynaptic or polysynaptic transmission. Most EPSPs (69/77) were significantly larger during the depolarized phase of the intracellular locomotor drive potential (LDP), and most IPSPs (35/46) were larger during the corresponding hyperpolarized phase. Bilateral cooling of the MedRF reversibly abolished locomotion of both hindlimbs as measured from the electroneurogram (ENG) activity of muscle nerves and simultaneously abolished or diminished the motoneuron PSPs and LDPs. Unilateral cooling of the VF blocked locomotion ipsilaterally and diminished it contralaterally with concomitant loss or decrease the motoneuron PSPs and LDPs. Relative to the side of motoneuron recording, cooling of the ipsilateral VF sometimes uncovered longer-latency EPSPs, whereas cooling of the contralateral VF abolished longer-latency EPSPs. It is concluded that MLR stimulation activates a pathway that relays in the MedRF and descends bilaterally in the VF to contact spinal interneurons that project to motoneurons. Local segmental pathways that activate or inhibit motoneurons during MLR-evoked fictive locomotion appear to be both ipsilateral and contralateral. PMID- 12634276 TI - Amplitude and frequency dependence of spike timing: implications for dynamic regulation. AB - The spike-time reliability of motoneurons in the Aplysia buccal motor ganglion was studied as a function of the frequency content and the relative amplitude of the fluctuations in the neuronal input, calculated as the coefficient of variation (CV). Measurements of spike-time reliability to sinusoidal and aperiodic inputs, as well as simulations of a noisy leaky integrate-and-fire neuron stimulated by spike trains drawn from a periodically modulated process, demonstrate that there are three qualitatively different CV-dependent mechanisms that determine reliability: noise-dominated (CV < 0.05 for Aplysia motoneurons) where spike timing is unreliable regardless of frequency content; resonance dominated (CV approximately 0.05-0.25) where reliability is reduced by removal of input frequencies equal to motoneuron firing rate; and amplitude-dominated (CV >0.35) where reliability depends on input frequencies greater than motoneuron firing rate. In the resonance-dominated regime, changes in the activity of the presynaptic inhibitory interneuron B4/5 alter motoneuron spike-time reliability. The increases or decreases in reliability occur coincident with small changes in motoneuron spiking rate due to changes in interneuron activity. Injection of a hyperpolarizing current into the motoneuron reproduces the interneuron-induced changes in reliability. The rate-dependent changes in reliability can be understood from the phase-locking properties of regularly spiking motoneurons to periodic inputs. Our observations demonstrate that the ability of a neuron to support a spike-time code can be actively controlled by varying the properties of the neuron and its input. PMID- 12634277 TI - Reduced synaptic plasticity in the lateral perforant path input to the dentate gyrus of aged C57BL/6 mice. AB - Hippocampal slices obtained from C57BL/6 mice (3-25 mo) were used to investigate the effects of aging on excitatory postsynaptic potentials (EPSPs) elicited in dentate gyrus with lateral perforant path stimulation. The maximal amplitude of the EPSP, as well as the degree of paired-pulse facilitation, was significantly reduced in animals aged 12 mo or more compared with younger animals (<12 mo). Although all animals showed equivalent short-term potentiation (STP) in response to high-frequency stimulation, this did not translate into a long-lasting increase in synaptic efficacy in the older animals. A significant degree of long term potentiation (LTP) of synaptic efficacy was only observed in animals <12 mo of age when measured 30 min after induction. Blocking GABAA-mediated inhibition significantly enhanced STP in younger and older animals; however, a significant degree of LTP was again only observed in slices taken from younger animals. These data indicate that the lateral perforant path input to the dentate gyrus is altered by the aging process, and that this results in a reduction in the capacity of this input to exhibit long-lasting synaptic plasticity. PMID- 12634278 TI - Human brain activation during sustained and intermittent submaximal fatigue muscle contractions: an FMRI study. AB - During prolonged submaximal muscle contractions, electromyographic (EMG) signals typically increase as a result of increasing motor unit activities to compensate for fatigue-induced force loss in the muscle. It is thought that cortical signals driving the muscle to higher activation levels also increases, but this has never been experimentally demonstrated. The purpose of this study was to quantify brain activation during submaximal fatigue muscle contractions using functional magnetic resonance imaging (fMRI). Twelve volunteers performed a sustained handgrip contraction for 225 s and 320 intermittent handgrip contractions ( approximately 960 s) at 30% maximal level while their brain was imaged. For the sustained contraction, EMG signals of the finger flexor muscles increased linearly while the target force was maintained. The fMRI-measured cortical activities in the contralateral sensorimotor cortex increased sharply during the first 150 s, then plateaued during the last 75 s. For the intermittent contractions, the EMG signals increased during the first 660 s and then began to decline, while the handgrip force also showed a sign of decrease despite maximal effort to maintain the force. The fMRI signal of the contralateral sensorimotor area showed a linear rise for most part of the task and plateaued at the end. For both the tasks, the fMRI signals in the ipsilateral sensorimotor cortex, prefrontal cortex, cingulate gyrus, supplementary motor area, and cerebellum exhibited steady increases. These results showed that the brain increased its output to reinforce the muscle for the continuation of the performance and possibly to process additional sensory information. PMID- 12634280 TI - Spike frequency adaptation and signaling properties of identified neurons in rodent deep spinal dorsal horn. AB - Using whole cell recordings, I analyzed the intrinsic discharge properties for 285 neurons in Rexed's laminae III-V of isolated hamster spinal cord preparations. Neurons were characterized by their responses to step-wise and ramp hold depolarizing current applied through the recording pipettes. Tonic cells (133/285; 47%) fired repetitively during step-wise current application. Firing decayed linearly (-0.14 to -4.3 imp . s(-1) . s(-1)) or was bimodal, with an initial exponential phase (tau approximately 450 ms) followed by a linear decline (-0.02 to -6.3 imp . s(-1) . s(-1)); discharge frequency was unrelated to current trajectory. Phasic-firing cells (108/285; 38%) responded with a burst discharge having an initial rapid, exponential decrease (tau approximately 30 ms) and subsequent linear decline (-1 to -78 imp . s(-1) . s(-1)). Phasic cells were activated preferentially by fast current ramps (slope, 70 pA/s-2.2 nA/s) with the number and frequency of impulses increasing with current slope. Delayed-firing cells (44/285; 15%), responded to current steps with an accelerating firing following a substantial latent period (0.5-4 s) and discharged during current ramps with slopes less than approximately 100 pA/s. Intracellular staining revealed a significant association between electrophysiological profile and neuronal morphology. A majority of presumed projection cells (22/30; 73%) exhibited tonic firing to step-wise activation. The preponderance of phasic and delayed firing cells, 93% (42/45) and 71% (12/17), respectively, were interneurons with local or intersegmental terminations. Differential sensitivity to static and time-varying components of membrane current suggest differences in neuronal signaling properties that may have important implications for integration of mechanosensory information in the deep spinal dorsal horn. PMID- 12634279 TI - TRPM8 mRNA is expressed in a subset of cold-responsive trigeminal neurons from rat. AB - Recent electrophysiological studies of cultured dorsal root and trigeminal ganglion neurons have suggested that multiple ionic mechanisms underlie the peripheral detection of cold temperatures. Several candidate "cold receptors," all of them ion channel proteins, have been implicated in this process. One of the most promising candidates is TRPM8, a nonselective cationic channel expressed in a subpopulation of sensory neurons that is activated both by decreases in temperature and the cooling compound menthol. However, evidence for the expression of TRPM8 in functionally defined cold-sensitive neurons has been lacking. Here, we combine fluorometric calcium imaging of cultured rat trigeminal neurons with single-cell RT-PCR to demonstrate that there are distinct subpopulations of cold responsive neurons and that TRPM8 likely contributes to cold transduction in one of them. TRPM8 is preferentially expressed within a subset of rapidly responsive, low-threshold (approximately 30 degrees C), cold sensitive neurons. A distinct class of slowly responsive cold-sensitive neurons that is activated at lower temperatures (approximately 20 degrees C) generally lacks detectable TRPM8 mRNA. Together with previous findings, our data support the notion that cold responsive neurons are functionally heterogeneous. PMID- 12634281 TI - Electrophysiological changes in adult rat dorsal horn neurons after neonatal peripheral inflammation. AB - Neonatal peripheral inflammation has been shown to produce profound anatomical changes in the dorsal horn of adult rats. In this study, we explored whether parallel physiological changes exist. Neonatal rats were injected with complete Freund's adjuvant (CFA) into the left hind paw. At 8-10 wk of age, single dorsal horn neurons were recorded in response to graded intensities of mechanical stimuli delivered to the receptive field. In addition, cord dorsum potentials, produced by electrical stimuli delivered to the left sciatic nerve at 2.5x threshold, were recorded bilaterally from L2 to S3. There were significant increases in background activity and responses to brush and pinch in neonatal rats that were treated with CFA, as compared with control rats. Further analysis showed similar significant changes when dorsal horn neurons were categorized into wide dynamic range (WDR), high-threshold (HT), and low-threshold (LT) groups. The receptive field was significantly larger in neonatally treated rats as compared with control rats. Additionally, there was a significant increase in the response to a 49 degrees C heat stimulus in neonatally treated rats as compared with control rats. There was also a trend for the amplitudes of N1, N2, and P waves of the cord dorsum potential to increase and latencies to decrease in neonatally treated rats, but no significant differences were detected between different levels of the spinal cord (L2 to S3). These data further support the notion that anatomical and physiological plasticity changes occurred in the spinal cord following early neonatal CFA treatment. PMID- 12634282 TI - Distinct firing properties of higher order thalamic relay neurons. AB - It has been proposed that the thalamus is composed of at least two types of nuclei. First-order relay nuclei transmit signals from the periphery to the cortex while higher order nuclei may route information from one cortical area to another. Although much is known about the functional properties of relay neurons in first-order nuclei, little is known about relay neurons belonging to higher order nuclei. We investigated the electrophysiological properties of relay cells in a higher-order thalamic nucleus using in vitro intracellular recordings from thalamic slices of the rat's lateral posterior nucleus (LPN). We found neurons of the LPN possess many of the same membrane properties as first-order relay neurons. These included low-threshold calcium spikes (IT) and burst firing, a mixed cation conductance (IH) that prevented membrane hyperpolarization, and a transient K+ conductance that delayed spike firing (IA). The repetitive firing characteristics of LPN neurons were more distinct. One group of cells, located in the more caudal regions of the LPN responded to depolarizing current pulses with a train of action potentials or in a regular spiking (RS) mode. This form of firing showed a steep but highly linear increase in firing frequency with increasing levels of membrane depolarization. Another group of cells, located in the more rostral regions of the LPN, responded to depolarizing current pulses with clusters of high-frequency bursts or in a clustered spiking (CS) mode. The overall firing frequency rose nonlinearly with membrane depolarization, but the frequency of a given burst remained relatively constant. The caudal LPN receives input from the superior colliculus, whereas the rostral LPN receives input from layers V and VI of the visual cortex. Thus the RS and CS cells may be driven by subcortical and cortical inputs respectively, and the distinct temporal properties of their response modes may be a necessary component of the LPN circuitry. PMID- 12634283 TI - Enhanced responses of the chorda tympani nerve to sugars in the ventromedial hypothalamic obese rat. AB - Sweet taste sensitivity in obese rats with lesions of the ventromedial hypothalamus (VMH) was studied by examining chorda tympani nerve responses to various taste stimuli including sugars. In the early progressive phase of obesity (2 wk after creating VMH lesions), there was no significant difference in the nerve responses to any taste stimulus between sham-operated and VMH-lesioned rats. In contrast, in the late phase of obesity (15-18 wk after VMH lesions), the magnitude of responses to sugars (except for fructose) was prominently greater than that in age-matched controls. High-fat diet-induced obese rats and streptozotocin-diabetic rats also showed greater chorda tympani nerve responses to sugars as was observed in VMH-lesioned obese rats, indicating that VMH lesions might not be specifically related to the enhanced gustatory neural responses to sugars. Although it has been demonstrated that the enhanced responses of the chorda tympani nerve to sugars in genetically diabetic db/db mice is largely attributable to the lack of the direct suppressive effect of leptin on the taste receptor cells, plasma leptin levels were not correlated with the changes in chorda tympani responsiveness to sugars in these models of obesity and diabetes. Accordingly, our results suggest that some chronic factors, including high blood glucose, inefficiency of insulin action, or leptin resistance may be related to the enhancement of chorda tympani nerve responses to sugars. PMID- 12634284 TI - Further evaluation of the basic nature of the human biological aging process based on a factor analysis of age-related physiological variables. AB - This study aimed to reexamine whether there exists a primary aging process that controls the rate of aging in a number of different functions. Eighty-six adult males who successively received a 2-day routine health checkup test for 7 years from 1992 to 1998 at the Kyoto Red Cross Hospital were selected as subjects. Nine candidate biomarkers of aging were selected from the 25 physiological variables based on the investigation of age-related changes. A principal factor analysis was applied to the partial correlation matrix for 9 selected biomarkers calculated by controlling for age. Furthermore, a confirmatory factor analysis in testing first- and second-order factor models was applied to the covariance matrix for 9 biomarkers. The results of these factor analyses revealed that there existed one general factor and three system-specific factors. Therefore, biological age changes can be viewed as a time-dependent complex integration of the primary and secondary aging processes. PMID- 12634285 TI - Age-related base excision repair activity in mouse brain and liver nuclear extracts. AB - To assess DNA repair activity relative to age, in vitro base excision repair assays were performed using brain and liver nuclear extracts prepared from mice of various ages. An 85% decline in repair activity was observed in brain nuclear extracts and a 50% decrease in liver nuclear extracts prepared from old mice compared with 6-day-old mice. Brain nuclear extracts prepared from old mice showed a decreased abundance of DNA polymerase-beta, but the addition of purified protein did not restore base excision repair activity. Abundances of other tested base excision repair proteins did not change relative to age. The conclusion is that, during aging, a decline in DNA repair could contribute to increased levels of DNA damage and mutagenesis. PMID- 12634287 TI - Discovering the secrets of successful longevity. PMID- 12634286 TI - Mortality and morbidity in laboratory-maintained Rhesus monkeys and effects of long-term dietary restriction. AB - Mortality and morbidity were examined in 117 laboratory-maintained rhesus monkeys studied over approximately 25 years (8 dietary-restricted [DR] and 109 ad libitum fed [AL] monkeys). During the study, 49 AL monkeys and 3 DR monkeys died. Compared with the DR monkeys, the AL monkeys had a 2.6-fold increased risk of death. Hyperinsulinemia led to a 3.7-fold increased risk of death (p <.05); concordantly, the risk of death decreased by 7%, per unit increase in insulin sensitivity (M). There was significant organ pathology in the AL at death. The age at median survival in the AL was approximately 25 years compared with 32 years in the DR. The oldest monkey was a diabetic female (AL) that lived to be 40 years of age. These results suggest that dietary restriction leads to an increased average age of death in primates, associated with the prevention of hyperinsulinemia and the mitigation of age-related disease. PMID- 12634288 TI - Distributions of ACE and APOE polymorphisms and their relations with dementia status in Korean centenarians. AB - BACKGROUND: Genetic polymorphisms of angiotensin-converting enzyme (ACE) and apolipoprotein E (APOE) have been reported to be associated with human longevity and dementia in the elderly. However, whether such putative longevity genes exert the same effects on different ethnic groups living in different environments is not well known. METHODS: We investigated the distributions of the ACE and APOE genotypes and their relations with dementia status in Korean centenarians by cross-sectional study. A total of 103 centenarians (13 men and 90 women, mean age 102.4 +/- 2.6 years) were included in this study. The allele frequencies of the genes were compared with those of two control groups: 7232 apparently healthy adults (4100 men and 3132 women) of mean age 48.5 +/- 9.6 years for the ACE genotyping, and 6435 adults (5008 men and 1427 women) of mean age 50.7 +/- 7.9 years for the APOE genotyping. The dementia status of the centenarians was assessed by clinical psychologist using the Clinical Dementia Rating (CDR) score. RESULTS: The frequencies of genotypes and alleles of the ACE and APOE genes of the centenarians were not significantly different from those of the control groups. There was a lack of association between presence of the D allele on the ACE gene and dementia status. However, the frequency of the epsilon4 allele of the APOE gene was significantly higher in centenarians with dementia than in centenarians without definitive dementia (9.1% versus 1.5%, p <.05). CONCLUSIONS: These results suggest that neither the ACE nor the APOE gene is significantly associated with longevity in the Korean population, but that the APOE epsilon4 allele is still related with dementia even at age 100 and older. PMID- 12634290 TI - Improving nursing home quality assessment: capturing the voice of cognitively impaired elders. PMID- 12634289 TI - Morbidity profiles of centenarians: survivors, delayers, and escapers. AB - BACKGROUND: The compression of morbidity hypothesis predicts that, in order to achieve their extreme old age, centenarians markedly delay or even escape diseases that would otherwise be lethal at younger ages. Phenotypic studies have not adequately characterized the prevalence and timing of age-related illnesses among those who achieve exceptional old age. Thus, we conducted a retrospective cohort study of centenarians to explore the timing of such diseases among centenarians. METHODS: Health history questionnaires were completed by 424 centenarians (aged 97-119 years) or their proxies. Lifetime (to-date) diagnoses of 10 major lethal illnesses (hypertension, heart disease, diabetes, stroke, nonskin cancer, skin cancer, osteoporosis, thyroid condition, Parkinson's disease, and chronic obstructive pulmonary disease) and one ocular disease (cataracts) that befall the elderly population, approximate age of diagnosis, level of alcohol and tobacco use, and presence or absence of cognitive impairment were assessed. Because of the retrospective nature of the study, the typically imprecise age of onset of cognitive impairment negated the ability to include age of onset of cognitive impairment in this aspect of the analyses. RESULTS: Examining the ages of onset for the 10 age-associated diseases and excluding cognitive impairment, we found that the centenarians fit into three morbidity profiles-Survivors, Delayers, and Escapers. 24% of male subjects and 43% of female subjects fit the Survivor profile, or those who had a diagnosis of an age associated illness prior to the age of 80. Delayers were individuals who delayed the onset of age-associated illness until at least the age of 80, and 44% of male and 42% of female subjects fit this profile. Escapers were individuals who attained their 100th year of life without the diagnosis of common age-associated illnesses, and 32% of male and 15% of female subjects fit the Escaper profile. When examining only the most lethal diseases of the elderly population, heart disease, nonskin cancer, and stroke, we found that 87% of male and 83% of female subjects delayed or escaped these diseases. Subjects with and without cognitive impairment did not differ in terms of the profile to which they belonged. CONCLUSIONS: These results suggest there may be multiple routes to achieving exceptional longevity and that there are sex differences according to which route is taken. These routes represent different phenotypes and thus likely different genotypes of centenarians. The identification of three types of centenarians, Survivors, Delayers, and Escapers, provides direction for future study into the factors that determine exceptional longevity. PMID- 12634292 TI - Depression in late life: review and commentary. AB - Depression is perhaps the most frequent cause of emotional suffering in later life and significantly decreases quality of life in older adults. In recent years, the literature on late-life depression has exploded. Many gaps in our understanding of the outcome of late-life depression have been filled. Intriguing findings have emerged regarding the etiology of late-onset depression. The number of studies documenting the evidence base for therapy has increased dramatically. Here, I first address case definition, and then I review the current community- and clinic-based epidemiological studies. Next I address the outcome of late-life depression, including morbidity and mortality studies. Then I present the extant evidence regarding the etiology of depression in late life from a biopsychosocial perspective. Finally, I present evidence for the current therapies prescribed for depressed elders, ranging from medications to group therapy. PMID- 12634291 TI - Quality of life measures for nursing home residents. AB - BACKGROUND: Quality of life (QOL) is a goal for nursing home residents, but measures are needed to tap this phenomenon. METHODS: In-person QOL interviews were attempted for 1988 residents, stratified by cognitive functioning, from 40 nursing homes in five states. Likert-type response options were used with reversion to dichotomous responses when necessary; z-score transformations were used to combine the formats. Tests of internal consistency and confirmatory factor analysis were performed; cluster analysis was used to shorten the scales. Correlations between domain scores were examined, and tests of convergent validity performed. Analyses were repeated for subgroups based on cognitive functioning levels. RESULTS: Long QOL scales were constructed for 1316 of the 1988 residents, including many with substantial cognitive impairment. Confirmatory factor analysis confirmed 10 QOL domains. Cronbach alphas ranged from.76 to.52. The majority (93%) of the 45 possible interscale correlations among domains were below.l4 and the rest were between.4 and.5. QOL scales were correlated with, but distinct from, residents' emotions ratings and overall satisfaction, and each was correlated with a corresponding summary rating for the domain. CONCLUSIONS: QOL can be feasibly measured from resident self-report for much of the nursing home population, including cognitively impaired residents. Additional research is suggested on the measures, but the approach has promise for regulation, continuous quality improvement, and public information. PMID- 12634293 TI - Effects of exercise training and hormone replacement therapy on lean and fat mass in postmenopausal women. AB - BACKGROUND: Menopause is associated with decreases in lean mass and increases in fat mass. Serum hormone levels and hormone replacement therapy (HRT) may modify the effects of exercise training on body composition in postmenopausal women. METHODS: We assessed the changes in total body and regional lean soft tissue and fat mass (using dual-energy x-ray absorptiometry) in 94 sedentary postmenopausal women, aged 40-65 years, after 12 months of resistance and weight-bearing aerobic exercise training. Women currently on oral HRT (n = 39) and not on HRT (n = 55) were randomized within groups to exercise and no exercise, resulting in four groups: exercise + HRT (n = 20), HRT (n = 22), exercise (n = 24), and control (n = 28). Fasting blood samples were measured for resting serum total levels of estrone, estradiol, cortisol, androstenedione, growth hormone, and insulin-like growth factor 1 at baseline and 12 months. RESULTS: We found significant effects of exercise on increases in total body, arm, and leg lean soft tissue mass, and decreases in leg fat mass and percentage of body fat. There were no interaction effects of exercise and HRT on the changes in muscle strength and body composition. No significant changes in total hormone levels were found after 12 months. CONCLUSIONS: Exercise training resulted in significant beneficial changes in lean soft tissue and fat mass in early postmenopausal women. These changes in body composition were neither influenced by prolonged HRT use nor accompanied by changes in total levels of the hormones determined in this study. PMID- 12634294 TI - Medical staff's decision-making process in the nursing home. AB - BACKGROUND: This paper describes the medical decision-making process at the time of status change events in the nursing home. METHODS: Six male physicians and 3 female nurse practitioners completed questionnaires that described the medical decision-making process for 70 residents of a large nonprofit nursing home. RESULTS: Hospitalization was the most frequently cited treatment considered and chosen; family members were involved in 39% of decisions, and nurses were involved in 34%. The most important considerations in making a decision were reported to be the resident's quality of life, the relative effectiveness of the treatment options, and the family's wishes. The levels of importance ascribed to the considerations were related to the physician's identity, specific resident characteristics (such as estimated life expectancy), and communication between the physician and resident (such as sharing knowledge of family wishes). CONCLUSIONS: The decision at the time of a status change event involves multiple conditions, multiple considerations, and multiple treatment options, and tends to result in either an active route, such as hospitalization, or a passive one, such as comfort care. The impact of the individual physician and the physician resident relationship on this process deserves further investigation. PMID- 12634295 TI - Embryonic stem cells: new possible therapy for degenerative diseases that affect elderly people. AB - The capacity of embryonic stem (ES) cells for virtually unlimited self renewal and differentiation has opened up the prospect of widespread applications in biomedical research and regenerative medicine. The use of these cells would overcome the problems of donor tissue shortage and implant rejection, if the cells are made immunocompatible with the recipient. Since the derivation in 1998 of human ES cell lines from preimplantation embryos, considerable research is centered on their biology, on how differentiation can be encouraged toward particular cell lineages, and also on the means to enrich and purify derivative cell types. In addition, ES cells may be used as an in vitro system not only to study cell differentiation but also to evaluate the effects of new drugs and the identification of genes as potential therapeutic targets. This review will summarize what is known about animal and human ES cells with particular emphasis on their application in four animal models of human diseases. Present studies of mouse ES cell transplantation reveal encouraging results but also technical barriers that have to be overcome before clinical trials can be considered. PMID- 12634296 TI - Inhibition of spermatogonial differentiation by testosterone. PMID- 12634297 TI - Epididymosomes and prostasomes: their roles in posttesticular maturation of the sperm cells. PMID- 12634298 TI - Cryptorchidism: incidence, risk factors, and potential role of environment; an update. PMID- 12634300 TI - Cloning--a matter of life. PMID- 12634299 TI - Comment on: Cryptorchidism: incidence, risk factors, and potential role of environment; an update. PMID- 12634301 TI - Diagnosis of nonobstructive azoospermia: the laboratory perspective. PMID- 12634302 TI - Management of psychogenic anejaculation. PMID- 12634303 TI - Expression of testicular germ cell genes identified by differential display analysis. AB - Using differential display reverse transcriptase-polymerase chain reaction (DDRT PCR) we identified transcripts encoding for the RNA helicase mDEAH9, Ran binding protein 5 (RanBP5), and 3 novel complementary DNAs designated GC3, GC12, and GC14 in developing testicular germ cells. Sources of RNA for the initial DDRT-PCR screen were purified mouse type A spermatogonia, adult mouse wild-type testis, and W/W(v) mutant mouse testis. We identified cDNA fragments for mDEAH9, RanBP5, GC3, GC12, and GC14 in testis and type A spermatogonia samples from wild-type mice, but not in samples from the W/W(v) mouse testis. These same transcripts were absent in Northern blots of testis RNA from mice treated with busulfan 30 days prior, but were present in testis RNA from wild-type mice at 5, 15, 25, and 40 days of age. The mDEAH9 gene was expressed in many tissues, whereas RanBP5 and GC12 genes were expressed predominantly in the testis with much lower expression in other tissues. The expression of GC3 and GC14 were limited to the testis as evidenced by Northern blot and RT-PCR analyses. The mDEAH9 transcript was not detected in cultured interstitial cells but was found at low levels in cultured immature Sertoli cells, whereas the RanBP5, GC3, GC12, and GC14 transcripts were not detected in either cultured testicular interstitial cells or cultured Sertoli cells. RT-PCR analyses of isolated spermatogonia, pachytene spermatocytes, and round spermatids revealed that mDEAH9, RanBP5, GC3, GC12, and GC14 genes were expressed in all 3 cellular populations. In situ hybridization analyses of testis samples from 40-day-old mice localized expression of mDEAH9, RanBP5, GC3, GC12, and GC14 to the seminiferous tubules. RanBP5 expression appeared to be regulated during the cycle of the seminiferous epithelium, with the highest expression in stages III through VII. Expression of GC14 was greatest in the meiotic germ cell populations. PMID- 12634304 TI - Balance of apoptosis and proliferation of germ cells related to spermatogenesis in aged men. AB - To clarify whether germ cell apoptosis is related to a decrease of germ cells in the aged testis with impaired spermatogenesis, we investigated the apoptotic rate of each germ cell type. Testicular specimens were obtained by orchiectomy from 36 men with advanced prostate cancer and by testicular biopsy from 21 men with obstructive azoospermia, which served as controls. The terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) technique was used to identify apoptosis. As a marker of cell proliferation activity, the expression of Ki-67 was immunohistochemically evaluated. Expression of Bcl-xl, which regulates apoptosis of germ cells, was also immunohistochemically examined. Histologically, except for spermatogonia, the ratios of primary spermatocytes, round spermatids, and elongated spermatids to Sertoli cells were significantly decreased in aged testes. The apoptotic rate in spermatogonia was significantly lower in aged men than it was in controls (0.11% +/- 0.06% vs 0.34% +/- 0.21%). Expression of Ki-67 in spermatogonia was decreased in aged men (18.6% +/- 6.0%) compared with that of controls (24.9% +/- 3.3%), suggesting that germ cell proliferation diminished with aging. Consequently, the balance of spermatogonial proliferation and apoptosis showed no difference between the two groups. This was believed to be one of reasons why spermatogonial numbers in aged testes was similar to those of controls. The apoptotic rate of primary spermatocytes in aged men was significantly elevated compared with that of controls (0.60% +/- 0.54% vs 0.22% +/-0.12%), resulting in a decrease of the number of primary spermatocytes per Sertoli cell. The expression of Bcl-xl was inversely correlated with the apoptotic rate in primary spermatocytes, suggesting that Bcl-xl may be related to the regulation of primary spermatocyte apoptosis. Based on these findings, we conclude that accelerated apoptosis of primary spermatocytes might account for a part of the mechanism of germ cell loss in aging men. PMID- 12634305 TI - Developmental and stage-specific expression of Smad2 and Smad3 in rat testis. AB - Members of the transforming growth factor beta type (TGFbeta) superfamily and their receptors are expressed in the testis, and are believed to play important paracrine and autocrine roles during testicular development and spermatogenesis. The Smad proteins are downstream mediators for the family of TGFbeta growth factors. Smad2 and Smad3 are associated with both TGFbeta and activin signaling. However, very little is known about the expression and regulation of the Smad signaling proteins in the testis. In the present study, we have determined that Smad2 and Smad3 proteins are expressed in the postnatal testes of rats from 5 days to 60 days of age. Expression levels for both proteins are higher in young rats than in sexually mature rats. Smad2 and Smad3 messenger RNA levels parallel protein expression. Smad2 and Smad3 proteins are mainly localized in the cytoplasm of meiotic germ cells, Sertoli cells, and Leydig cells. Smad3 protein is localized to the nucleus of preleptotene to zygotene primary spermatocytes in young rats. Both proteins are expressed throughout all stages of the cycle of seminiferous tubules but are expressed at their lowest levels at stages VII-VIII in the seminiferous epithelium of adult rats. The presence of these downstream mediators in these cell types supports a role for TGFbeta and activin during spermatogenesis. The difference between the expression of Smad2 and Smad3 suggests that they may have different functions within the testis. PMID- 12634306 TI - Editorial commentary: SMAD expression in the testis predicts age- and cell specific responses to activin and TGFbeta. PMID- 12634307 TI - Spermatogenetic expression of RNA-binding motif protein 7, a protein that interacts with splicing factors. AB - We have previously shown that a ubiquitously expressed RNA splicing factor, RNA binding motif 7 (RBM7), cloned from a testis complementary DNA library, enhances messenger RNA (mRNA) splicing in vitro and is expressed in a cell-restricted fashion. Herein, we detail its mRNA and protein expression in the rodent testis. RNA in situ hybridization shows that Rbm7 expression in rat germ cells closely parallels the entry and progression of meiosis. The expression commences in type B spermatogonia, it rises during the preleptotene stage, peaks in leptotene spermatocytes, and declines afterward, but increases again in stage-associated pachytene spermatocytes. An affinity-purified polyclonal antibody raised against a peptide corresponding to amino acids 202-224 of the mouse RBM7 recognized the predicted 35 kd protein both in testicular lysates and in in vitro translation reactions. Consistent with the in situ hybridization results, RBM7 immunoreactivity was also detected in type B spermatogonia, spanned the entire period of spermatocyte development, and extended to round and early elongated spermatids. Moreover, RBM7 appeared nuclear up to the mid pachytene stage and became cytoplasmic thereafter. Consistent with its role in RNA splicing, yeast 2 hybrid and glutathione S-transferase pull-down assays show that RBM7 interacts with splicing factor 3b subunit 2 (SAP145), and with the splicing regulator, SRp20. These interactions and the nuclear localization of RBM7 provide insights into its function in pre-mRNA processing in developing spermatocytes during entry into meiosis and progression through the meiotic prophase. PMID- 12634308 TI - Molecular analysis of the androgen receptor in ten prostate cancer specimens obtained before and after androgen ablation. AB - Hormonal or androgen-ablation (AA) therapy is the predominant form of systemic treatment for metastatic prostate cancer. Although an initial response to AA is observed in 70%-80% of patients with advanced disease, most tumors eventually progress to androgen-independent growth, and only a minority of affected individuals are alive 5 years following initiation of treatment. Because AA induces a dramatic change in the hormonal milieu of the patient and because these tumors maintain the ability to proliferate, it is possible that this treatment selects a population of cells with mutated androgen receptors (ARs) that sustain growth despite the absence of circulating androgen. To test this hypothesis we investigated the molecular structure of the AR in 10 prostate cancer specimens obtained before and after AA. Tumors (coded A through L) were microdissected to uniquely enrich genomic DNA from cancer cells. Exons 1-8 of the AR were screened by polymerase chain reaction, single-stranded conformational polymorphism, and sequence analysis. A mutation consisting of an expansion of the polyglutamine (poly-Q) repeat from 20 (found in 100% of the sequences of specimens obtained before AA) to 26 (found in 70% of the sequences of specimens obtained after AA) was detected in patient F. The 26 glutamine (Q26) AR readily translocated to the nucleus upon addition of androgen, and did not show significant differences in its ability to bind (3)[H]-dihydrotestosterone compared to its wild-type counterpart. Nevertheless, analysis of transcriptional activity showed that the Q66 AR was transcriptionally 30%-50% less active than the wild-type molecule. Because clones of AR with an expanded poly-Q tract were detected only in the specimen from patient F obtained after AA, we conclude that in specific circumstances, AA treatments can select variant forms of the AR in the prostate of patients affected by prostate cancer. Further experiments are needed to conclusively determine whether the Q26 clone was responsible for sustaining survival of prostate cancer cells in the androgen-depleted milieu of this patient. PMID- 12634309 TI - Seasonal variation and age-related changes in human semen parameters. AB - Although semen quality has been discussed extensively with regard to age and season in the andrology literature, the results vary and firm conclusions are still outstanding. To investigate seasonal and age-related variations in human semen parameters, we analyzed data that were collected from an andrology clinic population. We performed a retrospective review of 551 semen analysis records collected from 1989 to 2000 from the Vincent Memorial Andrology Laboratory at Massachusetts General Hospital. Semen volume, sperm concentration, total sperm count, motility, total motile sperm, and morphology significantly decreased as age increased. In addition, as age increased, the percentage of sperm with tail defects increased. Sperm concentration was significantly higher in winter (mean 157.9 million/mL) than in fall (mean 119.1 million/mL) (P <.05). The mean percentage of sperm with normal morphology was significantly higher in winter (9.2%) than in summer and spring (7.0% and 7.5%, respectively; P <.05). The mean percentage of sperm with head defects was significantly higher in fall and summer (74.0% and 72.3%, respectively) than in winter (68.6%; P <.05). Seasonal variations were found in sperm concentration and morphology, with higher sperm concentrations in winter than in fall, and a greater percentage of sperm with normal morphology in winter than in spring and summer. Sperm concentration was lowest in the fall, whereas the percentage of sperm with normal morphology was lowest in summer. Semen volume, sperm concentration, total sperm count, motility, total motile sperm, and morphology decreased as age increased. PMID- 12634310 TI - The human sperm head: a key for successful fertilization. AB - In order to examine the predictive value of determining the sperm head shape, the acrosomal size, the presence of acrosomal vacuoles, and the challenged acrosome reaction (AR) on the outcome of a standard in vitro fertilization (IVF) program, a prospective study was conducted that included 75 couples undergoing IVF treatment. An assessment of sperm morphology was performed using the Hobson Sperm Tracker (Hobson Tracker Limited, Sheffield, United Kingdom). The assessment of the AR was performed before and after adding pooled undiluted human follicular fluid (FF). The outcome measure was an IVF rate of inseminated oocytes. A positive correlation was found between the fertilization rate (FR%) and the proportion of the sperm with a normal (oval) head shape (P <.001), the sperm exhibiting acrosomal vacuoles (P <.003), the sperm with a normal acrosomal size (40%-70% of total head area, P <.025), and the sperm undergoing AR after adding FF (P <.001). Multiple logistic regression analysis revealed that by incorporating the above 4 parameters, the sensitivity of prediction of IVF FR% values was 79%, and the specificity was 93%, with a positive predictive value of 96%. This study shows that the multiparametric assessment of the sperm head is useful in predicting the FR% values of a standard IVF treatment. The automated analysis used in this study is shown to maintain a level of precision and accuracy acceptable for application in a routine semen analysis situation. PMID- 12634311 TI - Cavernous neurotomy causes hypoxia and fibrosis in rat corpus cavernosum. AB - The etiologies of erectile dysfunction (ED) after nerve-sparing radical prostatectomy have not been clearly elucidated. The aim of this study was to evaluate the effects of cavernous nerve injury on cavernous fibrosis, and to consider measures to prevent irreversible damage to the cavernous tissues. Twenty male Sprague-Dawley rats constituted the study population. The animals were divided into 2 groups; group 1 consisted of sham-operated rats (n = 10), and group 2 consisted of rats that underwent incision of both cavernous nerves (n = 10). Three months later, all rats underwent intracavernous papaverine injection (300 and 600 mg), and intracorporal pressures were recorded. Transforming growth factor-beta(1) (TGF-beta(1)) messenger RNA (mRNA) expression from rat penile tissue was measured using reverse transcriptase-polymerase chain reaction. Hypoxia-inducible factor-1alpha (HIF-1alpha), TGF-beta(1), and collagen I and III protein expressions were determined by Western blot analysis and immunohistochemical staining. Erectile function as studied with intracavernosal papaverine injection and histological analysis of penile cross-sections at 3 months was similar in both groups. TGF-beta(1) mRNA expression, HIF-1alpha, TGF beta(1), and collagen I and III protein expressions were significantly greater in the neurotomy group. Immunohistochemical staining for TGF-beta(1), HIF-1alpha, and collagen III were qualitatively more positive in the neurotomy group, whereas collagen I staining was similar. This study demonstrates an increase in TGF beta(1), HIF-1alpha, and collagen III synthesis in rat cavernosal smooth musculature after cavernous neurotomies. In theory, cavernous fibrosis may be reduced by employing various vasoactive agents or interventions that increase oxygenation to the corporal tissues during the postoperative period. PMID- 12634312 TI - Deterioration of plasma membrane is associated with activated caspases in human spermatozoa. AB - Spermatozoa with deteriorated plasma membranes can be separated by magnetic activated cell sorting (MACS) after binding superparamagnetic annexin V conjugated microbeads (ANMBs) to membrane phosphatidylserine (PS). Semen samples from 15 donors and 25 infertile patients were divided into 2 spermatozoal fractions by annexin V-MACS. Activated caspases (aCPs), which mediate degradations of cell quality, were determined by CaspaTag in the 2 subpopulations. Spermatozoa from donors showed lower levels of bound annexin V (3.6% +/- 0.5% vs 11.9% +/- 1.1%; P <.01) and aCPs (21.8% +/- 2.6% vs 43.2% +/- 2.1%; P <.01) than did spermatozoa from infertile patients. MACS resulted in a decrease of spermatozoa with aCPs from 21.8% +/- 2.6% (before separation) to 9.2% +/- 1.4% (in the ANMB-negative fraction) in donors and from 43.2% +/- 2.1% to 18.8% +/- 2.6% in infertile patients (mean +/- SEM; P <.01). Separation effects of the MACS technique were confirmed with flow cytometry using anti-annexin V antibodies and with electron microscopy. ANMB-MACS removes spermatozoa with PS bound annexin V and produces a higher quality spermatozoal fraction. Spermatozoa with a deteriorated membrane are characterized by an increase in aCPs. A higher percentage of spermatozoa with ANMBs bound to PS and with aCPs were found in infertile patients. PMID- 12634313 TI - Spontaneous DNA fragmentation in swim-up selected human spermatozoa during long term incubation. AB - The origin and the meaning of DNA fragmentation in ejaculated human spermatozoa are not yet clear, although some hypotheses have been proposed. In the present study, we used investigated sperm DNA fragmentation by terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end labeling (TUNEL)-coupled flow cytometry to investigate DNA fragmentation in spermatozoa that were selected by the swim-up procedure and incubated long-term. In addition, using flow cytometry we detected annexin V binding assay and propidium iodide staining, and we also studied membrane phosphatidylserine translocation and the loss of membrane integrity in the same sperm populations that we used in the TUNEL investigation. We found that in vitro sperm DNA fragmentation 1) occurs after ejaculation under experimental conditions without the involvement of any external factor, 2) is not affected by treatment with the nuclease inhibitor aurintricarboxylic acid, 3) is increased by treatment with the glutathione peroxidase inhibitor mercaptosuccinate, 4) correlates with basal values (ie, just after swim-up selection) of DNA fragmentation in teratozoospermic but not in normospermic semen samples, 5) develops in a sharply associated manner with the in vitro occurrence of sperm necrosis, and 6) is predicted by the basal value of annexin V binding in viable spermatozoa. These findings suggest an involvement of endogenously produced reactive oxygen species as the possible cause of in vitro sperm DNA fragmentation. PMID- 12634314 TI - Rapid proliferation of prostatic epithelial cells in spontaneously hypertensive rats: a model of spontaneous hypertension and prostate hyperplasia. AB - Spontaneously hypertensive rats (SHRs), a commonly used model of genetic hypertension, exhibit features of glandular hyperplasia of the ventral prostate, including the narrowing of acini with epithelial protrusions into the lumen and the piling up of epithelial cells. These rats also have frequent urinary voiding. In order to define the fundamental processes that lead to prostatic hyperplasia in SHRs, we compared the proliferation rate of their prostatic epithelial cells (PECs) in primary culture and in vivo to that of Wistar-Kyoto rats (WKYs), their normotensive controls. In vitro, primary cultures of SHR PECs had a shorter doubling time than those of WKY (3.3 vs 8.0 days) and showed higher levels of bromodeoxyuridine (BrdU) incorporation into DNA. In vivo, the BrdU incorporation seen 48 hours following injection was observed primarily in areas of epithelial piling up, which are seen in SHRs but not in WKYs. We concluded that prostate hyperplasia in SHRs results from a genuine increase in the proliferation rate of PECs and that this rapid proliferation is a fundamental feature of SHR PECs, maintained both in vivo and in vitro. Thus, SHRs can serve as a model for glandular hyperplasia of the prostate, resulting from a genetic tendency for an increased rate of cell proliferation. PMID- 12634316 TI - An increased CAG repeat length in the androgen receptor gene in azoospermic ICSI candidates. AB - The androgen receptor gene has a polymorphic trinucleotide repeat that encodes a polyglutamine tract in its N-terminal transactivation domain. We started this study in order to find out whether a correlation existed between the length of this polymorphic tract and the presence of azoospermia in candidates for intracytoplasmic sperm injection (ICSI). The CAG repeat length in exon 1 of the androgen receptor (AR) gene was directly sequenced in 102 patients with azoospermia and in 96 fertile controls. Hormone levels were also measured in patients with azoospermia. The mean AR gene CAG repeat length was significantly larger in azoospermic subjects than it was in control fertile men (23.25 +/- 2.7 versus 22.42 +/- 2.8; P =.033). A receiver operating characteristic analysis evidenced a cutoff point at 22/23 CAG repeats at which the probability of being azoospermic increased 2.2 times. Subsequent logistic regression analysis of the data showed that the odds for azoospermia increased with the number of CAG repeats. Men with more than 26 CAG repeats have a 4.09 greater risk of being azoospermic. Therefore, in our candidates for ICSI, a direct correlation exists between the CAG repeat length in the exon 1 of the AR gene and the risk of being azoospermic. PMID- 12634315 TI - Neoplastic potential of germ cells in relation to disturbances of gonadal organogenesis and changes in karyotype. AB - The study consisted of 46 intersexual patients who underwent gonadectomy at the age of 3 months to 19 years because of gonadal dysgenesis (GD; 40 cases) or true hermaphroditism (bisexual gonads; 6 cases). In patients with GD, the incidence of the 46,XY karyotype was 67.5%, whereas the remaining patients exhibited numerical and structural aberrations of sex chromosomes (NSASs), and all patients with bisexual gonads revealed NSAS. Seminoma was diagnosed in 1 patient with the 46,XY karyotype and pure GD (streak gonads). Intratubular carcinoma in situ (CIS) appeared as an exclusive lesion in 61.5% of 13 patients with mixed GD, in 54% of 11 patients with partial GD (bilateral testes), in 16.7% of 6 patients with bisexual gonads, and in none of 13 patients with pure GD. CIS also appeared in tubules in the vicinity of sex cord-derived tumors (gonadoblastoma nests and unclassified mixed germ cell-sex cord-stromal tumor; MGCSCST) and within the tumors. In 3 patients, gonadoblastoma replaced the whole bilateral gonads and is referred to as gonadoblastoma-only GD. The incidence of neoplastic lesions (mostly bilateral) was 90.9% in patients with partial GD, 76.9% (mostly unilateral) in patients with mixed GD, 23.1% (unilateral) in patients with pure GD, and 16.7% (unilateral) in patients with bisexual gonads. Disregarding types of disturbances of gonadal organogenesis, the incidence of lesions was 71.4% in 28 patients with the 46,XY karyotype and 35.3% in 17 patients with NSAS. We conclude, first, that NSAS is not a prerequisite for the appearance of GD and GD is more frequently associated with the 46,XY karyotype. Second, the spectrum of germ cell neoplastic lesions in GD is wider than reported. Besides germ cell carcinoma, CIS, and gonadoblastoma nests, the spectrum also includes a tumor of gonadoblastoma-only in cases of GD and MGCSCST. Third, the incidence of neoplastic lesions is related more to the severity of the disturbances of gonadal organogenesis than it is to aberrations in sex chromosomes. Fourth, less disturbed testicular organogenesis predisposes these patients more toward germ cell neoplastic lesions, which suggests that the testicular environment of a dysgenetic gonad plays an important role in germ cell neoplasia initiation, maintenance, or both. PMID- 12634317 TI - Underestimation of subfertility among relatives when using a family history: taboo bias. AB - Family history is widely used in clinical practice and research in order to study genetic aspects of disorders in general, and is recommended as a tool in the assessment of male subfertility. Unfortunately, little is known about the validity of this tool. In this survey, we sent questionnaires to 474 randomly selected men aged 25-40 years in order to collect data on subfertility among them and their relatives. A nonresponder study was also conducted in order to evaluate selection bias. A personal interview was also performed with some respondents in order to gauge how well the data corresponded with questionnaires that were returned. Two hundred forty-three men (51.3%) completed the questionnaire. The responders reported a significantly lower prevalence of subfertility among their relatives than among themselves. Among brothers, the reported prevalence was about 5 times lower (ie, 3.6%) than among responders (15.3%). The nonresponder study and personal interviews showed that these differences were not caused by a selective response to the survey or by the use of a questionnaire instead of a personal interview. We conclude that subfertility among relatives is severely underestimated through the use of family history, probably because of the taboo of discussing subfertility. Knowledge of subfertility may spread selectively within families, causing substantial misclassification. Therefore, researchers and clinicians should be aware that an inquiry of family history is likely to lead to underestimation of subfertility among relatives. PMID- 12634318 TI - Antibody-based assay for N-deacetylase activity of heparan sulfate/heparin N deacetylase/N-sulfotransferase (NDST): novel characteristics of NDST-1 and -2. AB - A new assay was developed to measure the N-deacetylase activity of the glucosaminyl N-deacetylase/N-sulfotransferases (NDSTs), which are key enzymes in sulfation of heparan sulfate (HS)/heparin. The assay is based on the recognition of NDST-generated N-unsubstituted glucosamine units in Escherichia coli K5 capsular polysaccharide or in HSs by monoclonal antibody JM-403. Substrate specificity and potential product inhibition of the NDST isoforms 1 and 2 were analyzed by comparing lysates of human 293 kidney cells stably transfected with mouse NDST-1 or -2. We found HSs to be excellent substrates for both NDST enzymes. Both NDST-1 and -2 N-deacetylate heparan sulfate from human aorta ( approximately 0.6 sulfate groups/disaccharide) with comparable high efficiency, apparent Km values of 0.35 and 0.76 microM (calculation based on [HexA]) being lower (representing a higher affinity) than those for K5 polysaccharide (13.3 and 4.7 microM, respectively). Comparison of various HS preparations and the unsulfated K5 polysaccharide as substrates indicate that both NDST-1 and -2 can differentially N-sulfate polysaccharides already modified to some extent by various other enzymes involved in HS/heparin synthesis. Both enzymes were equally inhibited by N-sulfated sequences (>or=6 sugar residues) present in N-sulfated K5, N-deacetylated N-resulfated HS, and heparin. Our primary findings were confirmed in the conventional N-deacetylase assay measuring the release of 3H acetate of radiolabeled K5 or HS as substrates. We furthermore showed that NDST N deacetylase activity in crude cell/tissue lysates can be partially blocked by endogenous HS/heparin. We speculate that in HS biosynthesis, some NDST variants initiate HS modification/sulfation reactions, whereas other (or the same) NDST isoforms later on fill in or extend already modified HS sequences. PMID- 12634319 TI - All in the family: the UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferases. AB - Mucin-type linkages (GalNAcalpha1-O-Ser/Thr) are initiated by a family of glycosyltransferases known as the UDP-N-acetylgalactosamine:polypeptide N acetylgalactosaminyltransferases (ppGaNTases, EC 2.4.1.41). These enzymes transfer GalNAc from the sugar donor UDP-GalNAc to serine and threonine residues, forming an alpha anomeric linkage. Despite the seeming simplicity of ppGaNTase catalytic function, it is estimated on the basis of in silico analysis that there are 24 unique ppGaNTase human genes. ppGaNTase isoforms display tissue-specific expression in adult mammals as well as unique spatial and temporal patterns of expression during murine development. In vitro assays suggest that a subset of the ppGaNTases have overlapping substrate specificities, but at least two ppGaNTases (ppGaNTase-T7 and -T9 [now designated -T10]) appear to require the prior addition of GalNAc to a synthetic peptide before they can catalyze sugar transfer to this substrate. Site-specific O-glycosylation by several ppGaNTases is influenced by the position and structure of previously added O-glycans. Collectively, these observations argue in favor of a hierarchical addition of core GalNAc residues to the apomucin. Various forms of O-glycan pathobiology may be reexamined in light of the existence of an extensive ppGaNTase family of enzymes. Recent work has demonstrated that at least one ppGaNTase isoform is required for normal development in Drosophila melanogaster. Structural insights will no doubt lead to the development of isoform-specific inhibitors. Such tools will prove valuable to furthering our understanding of the functional roles played by O-glycans. PMID- 12634320 TI - Carbohydrate-binding properties of human neo-CRP and its relationship to phosphorylcholine-binding site. AB - Binding characteristics of two types of ligands for human neo-C-reactive protein (neo-CRP), which is a conformationally altered but physiologically relevant form of CRP, were studied fluorometrically by probing CRP immobilized on a polystyrene surface with europium-labeled ligands. Two Eu-ligands used were bovine serum albumin derivatives that contain on average 40 residues of ligand structures, one derivative containing phosphorylcholine (PC) and the other lactosyl residues. The PC-containing ligands required the presence of calcium for binding, whereas galactose-containing derivatives bound in the absence of calcium. The optimal pH for the PC-dependent binding was broad (pH 6-8), whereas the best binding pH for the galactose-dependent binding was around 6. The carbohydrate-mediated binding is rather nonspecific: the binding site prefers galactose configuration, but other hexoses can be accommodated. The two best monosaccharide inhibitors at this site were galactose-6-phosphate and galacturonic acid, suggesting the importance of having a negatively charged group at C-6 position of galactose. In fact, the phosphate-binding site is common to both PC and sugar phosphates, and the choline and the sugar-binding sites are probably located on either side of the phosphate binding site. Binding characteristics of Eu-labeled PC-BSA to neo-CRP are quite similar to that found for native CRP in solution phase [Lee et al. (2002) J. Biol. Chem., 277, 225-232], whereas binding of sugar phosphates by neo-CRP shows considerably less stringent requirements compared to native CRP. For instance, galactose-alpha1-phosphate was not inhibitory at all in the native CRP binding assay, whereas it was a good inhibitor in the neo-CRP assay. PMID- 12634321 TI - Complex-type biantennary N-glycans of recombinant human transferrin from Trichoplusia ni insect cells expressing mammalian [beta]-1,4 galactosyltransferase and [beta]-1,2-N-acetylglucosaminyltransferase II. AB - A novel recombinant baculovirus expression vector was used to produce His-tagged human transferrin in a transformed insect cell line (Tn5beta4GalT) that constitutively expresses a mammalian beta-1,4-galactosyltransferase. This virus encoded the His-tagged human transferrin protein in conventional fashion under the control of the very late polyhedrin promoter. In addition, to enhance the synthesis of galactosylated biantennary N-glycans, this virus encoded human beta 1,2- N-acetylglucosaminyltransferase II under the control of an immediate-early (ie1) promoter. Detailed analyses by MALDI-TOF MS, exoglycosidase digestion, and two-dimensional HPLC revealed that the N-glycans on the purified recombinant human transferrin produced by this virus-host system included four different fully galactosylated, biantennary, complex-type glycans. Thus, this study describes a novel baculovirus-host system, which can be used to produce a recombinant glycoprotein with fully galactosylated, biantennary N-glycans. PMID- 12634322 TI - Glycosaminoglycan structures required for strong binding to midkine, a heparin binding growth factor. AB - Midkine (MK), a heparin-binding growth factor, binds strongly to oversulfated structures in chondroitin sulfates (CSs) and heparan sulfate. To elucidate the carbohydrate structure actually involved in the strong binding, dissected brains from 13-day mouse embryos were incubated with [14C]-glucosamine. The labeled glycosaminoglycans were fractionated by MK-agarose affinity chromatography to a weakly binding fraction, which was eluted by 0.5 M NaCl, and a strongly binding fraction, which was eluted by higher NaCl concentrations. Among the unsaturated disaccharides released from the strongly binding fraction by chondroitinase ABC, DeltaDi-diSE with 4,6-disulfated N-acetylgalactosamine accounted for 32.3%, whereas its content was lower in the weakly binding fraction. Artificial CS-E structure was formed using N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase purified from squid or recombinant human enzyme. Analysis of the products and their interaction with MK revealed that E units without 3-O-sulfation of glucuronic acid are sufficient for strong binding, provided that they are present as a dense cluster. Among the sulfated disaccharides released by heparitinase digestion, the trisulfated one, DeltaDiHS-triS, was the most abundant in the strongly binding fraction and was lower in the weakly binding fraction. Together with results of previous studies, we concluded that the multivalent trisulfated heparin-like unit is another structure involved in strong binding to MK. PMID- 12634323 TI - Five Lec1 CHO cell mutants have distinct Mgat1 gene mutations that encode truncated N-acetylglucosaminyltransferase I. AB - Lec1 CHO cell mutants lack N-acetylglucosaminyltransferase I (GlcNAc-TI) activity and do not synthesize complex or hybrid N-glycans. The origins of six independent lec1 mutations are shown to reside in the coding region of the Mgat1 gene, proving that GlcNAc-TI is mutated in Lec1 mutants. One mutant has Mgat1 gene transcripts of reduced size, whereas the others possess transcripts of approximately normal size and amount containing a unique insertion or transition mutation that leads to a premature stop codon in the Mgat1 gene coding region. The lec1 mutation in the Lec3.2.8.1 mutant, a line used to generate minimally glycosylated membrane glycoproteins for X-ray crystallography, is a G insertion that leads to a nonsense codon after amino acid 391. The Pro-Lec1.3C line from the ATCC and in laboratory stocks, a line used widely for diverse purposes, possesses a C insertion in the Mgat1 gene coding exon, causing a frame shift and producing a stable, truncated approximately 24-kDa product. Mgat1 gene mutations were confirmed by sequencing genomic DNA PCR products. Mutant cDNAs were reverted by site-directed mutagenesis and shown to confer wild-type lectin binding and GlcNAc-TI activity on Lec1 transfectants. Surprisingly, three Mgat1 gene nucleotide changes previously reported in Pro-Lec1.3C cells (Puthalakath et al. [1996] J. Biol. Chem., 271, 27818-27822) were not detected in this study. These Lec1 mutants provide a novel cohort for investigating the effects on Golgi trafficking and kin recognition of deletion mutants of GlcNAc-TI expressed at endogenous rather than nonphysiological levels. PMID- 12634324 TI - Moving folded proteins across the bacterial cell membrane. AB - The Tat protein export system is located in the bacterial cytoplasmic membrane and operates in parallel to the well-known Sec pathway. While the Sec system only transports unstructured substrates, the function of the Tat pathway is to translocate folded proteins. The Tat translocase thus faces the formidable challenge of moving structured macromolecular substrates across the bacterial cytoplasmic membrane without rendering the membrane freely permeable to protons and other ions. The substrates of the Tat pathway are often proteins that bind cofactor molecules in the cytoplasm, and are thus folded, prior to export. Such periplasmic cofactor-containing proteins are essential for most types of bacterial respiratory and photosynthetic energy metabolism. In addition, the Tat pathway is involved in outer membrane biosynthesis and in bacterial pathogenesis. Substrates are targeted to the Tat pathway by amino-terminal signal sequences harbouring consecutive, essentially invariant, arginine residues, and movement of proteins through the Tat system is energized by the transmembrane proton electrochemical gradient. The TatA protein probably forms the transport channel while the TatBC proteins act as a receptor complex that recognizes the signal peptide of the substrate protein. PMID- 12634325 TI - Beginnings of microbiology and biochemistry: the contribution of yeast research. AB - With improvements in microscopes early in the nineteenth century, yeasts were seen to be living organisms, although some famous scientists ridiculed the idea and their influence held back the development of microbiology. In the 1850s and 1860s, yeasts were established as microbes and responsible for alcoholic fermentation, and this led to the study of the role of bacteria in lactic and other fermentations, as well as bacterial pathogenicity. At this time, there were difficulties in distinguishing between the activities of microbes and of extracellular enzymes. Between 1884 and 1894, Emil Fischer's study of sugar utilization by yeasts generated an understanding of enzymic specificity and the nature of enzyme-substrate complexes. PMID- 12634326 TI - The extracytoplasmic folding factor PrsA is required for protein secretion only in the presence of the cell wall in Bacillus subtilis. AB - Pulse-chase labelling was used to study the role of the cell wall microenvironment in the functioning of Bacillus subtilis PrsA, an extracellular lipoprotein and member of the parvulin family of peptidylprolyl cis/trans isomerases. It was found that in protoplasts, and thus in the absence of a cell wall matrix, the post-translocational folding, stability and secretion of the AmyQ alpha-amylase were independent of PrsA, in contrast to the strict dependency found in rods. The results indicate that PrsA is dedicated to assisting the folding and stability of exported proteins in the particular microenvironment of the cytoplasmic membrane-cell wall interface, possibly as a chaperone preventing unproductive interactions with the wall. The data also provide evidence for a crucial role of the wall in protein secretion. The presence of the wall directly or indirectly facilitates the release of AmyQ from the cell membrane and affects the rate of the signal peptide processing. PMID- 12634327 TI - Haemin uptake and use as an iron source by Candida albicans: role of CaHMX1 encoded haem oxygenase. AB - Candida albicans, unlike Saccharomyces cerevisiae, was able to use extracellular haemin as an iron source. Haemin uptake kinetics by C. albicans cells showed two phases: a rapid phase of haemin binding (with a K(d) of about 0.2 microM) followed by a slower uptake phase. Both phases were strongly induced in iron deficient cells compared to iron-rich cells. Haemin uptake did not depend on the previously characterized reductive iron uptake system and siderophore uptake system. CaHMX1, encoding a putative haem oxygenase, was shown to be required for iron assimilation from haemin. A double DeltaCahmx1 mutant was constructed. This mutant could not grow with haemin as the sole iron source, although haemin uptake was not affected. The three different iron uptake systems (reductive, siderophore and haemin) were regulated independently and in a complex manner. CaHMX1 expression was induced by iron deprivation, by haemin and by a shift of temperature from 30 to 37 degrees C. CaHMX1 expression was strongly deregulated in a Deltaefg1 mutant but not in a Deltatup1 mutant. C. albicans colonies forming on agar plates with haemin as the sole iron source showed a very unusual morphology. Colonies were made up of tubular structures that were organized into a complex network. The effect of haemin on filamentation was increased in the double DeltaCahmx1 mutant. This study provides the first experimental evidence that haem oxygenase is required for iron assimilation from haem by a pathogenic fungus. PMID- 12634328 TI - Characterization, localization and functional analysis of Gpr1p, a protein affecting sensitivity to acetic acid in the yeast Yarrowia lipolytica. AB - Adaptation of cells to acetic acid requires a hitherto unknown number of proteins. Studies on the GPR1 gene and its encoded protein in the ascomycetous fungus Yarrowia lipolytica have revealed an involvement of this protein in the molecular processes of adaptation to acetic acid. Gpr1p belongs to a novel family of conserved proteins in prokaryotic and eukaryotic organisms that is characterized by the two motifs (A/G)NPAPLGL and SYG(X)FW (GPR1_FUN34_YaaH protein family). Analysis of four trans-dominant mutations and N-terminal deletion analysis of Gpr1p identified the amino acid sequence FGGTLN important for function of this protein in Y. lipolytica. Deletion of GPR1 slowed down adaptation to acetic acid, but had no effect on growth in the presence of acetic acid. Expression of GPR1 is induced by acetic acid and moderately repressed by glucose. It was shown by subcellular fractionation that Gpr1p is an integral membrane protein, which is also suggested by the presence of five to six putative transmembrane spanning regions. Fluorescence microscopy confirmed a localization to the plasma membrane. A model is presented describing a hypothetical function of Gpr1p during adaptation to acetic acid. PMID- 12634329 TI - Reconstruction of C(3) and C(4) metabolism in Methylobacterium extorquens AM1 using transposon mutagenesis. AB - The growth of Methylobacterium extorquens AM1 on C(1) compounds has been well studied, but little is known about how this methylotroph grows on multicarbon compounds. A Tn5 transposon mutagenesis procedure was performed to identify genes involved in the growth of M. extorquens AM1 on succinate and pyruvate. Of the 15000 insertion colonies screened, 71 mutants were found that grew on methanol but either grew slowly or were unable to grow on one or both of the multicarbon substrates. For each of these mutants, the chromosomal region adjacent to the insertion site was sequenced, and 55 different genes were identified and assigned putative functions. These genes fell into a number of predicted categories, including central carbon metabolism, carbohydrate metabolism, regulation, transport and non-essential housekeeping functions. This study focused on genes predicted to encode enzymes of central heterotrophic metabolism: 2-oxoglutarate dehydrogenase, pyruvate dehydrogenase and NADH : ubiquinone oxidoreductase. In each case, the mutants showed normal growth on methanol and impaired growth on pyruvate and succinate, consistent with a role specific to heterotrophic metabolism. For the first two cases, no detectable activity of the corresponding enzyme was found in the mutant, verifying the predictions. The results of this study were used to reconstruct multicarbon metabolism of M. extorquens AM1 during growth on methanol, succinate and pyruvate. PMID- 12634330 TI - Capacity of ivanolysin O to replace listeriolysin O in phagosomal escape and in vivo survival of Listeria monocytogenes. AB - Listeriolysin O (LLO, hly-encoded) is a major virulence factor secreted by the pathogen Listeria monocytogenes. The amino acid sequence of LLO shows a high degree of similarity with that of ivanolysin O (ILO), the cytolysin secreted by the ruminant pathogen Listeria ivanovii. Here, it was tested whether ILO could functionally replace LLO by expressing the gene encoding ILO under the control of the hly promoter, in an hly-deleted strain of L. monocytogenes. It is shown that ILO allows efficient phagosomal escape of L. monocytogenes in both macrophages and hepatocytes. Moreover, expression of ILO is not cytotoxic and promotes normal intracellular multiplication. In vivo, the ILO-expressing strain can multiply and persist for several days in the liver of infected mice but is unable to survive in the spleen. This work underscores the key role played by the cytolysin in the virulence of pathogenic Listeria. PMID- 12634331 TI - Phase variation in Bartonella henselae. AB - Bartonella henselae is a fastidious, Gram-negative bacterial pathogen of cats and humans. Previous workers have shown that serial passage in vitro leads to attenuation of virulence-associated attributes such as expression of pili, invasion of human epithelial cell lines and the stimulation of endothelial cell proliferation. In contrast to the published data, it was found that pilin expression is frequently preserved in organisms which have undergone phase variation in vitro. Transition from a slow-growing, dry agar-pitting (DAP) to a faster-growing, smooth non-agar-pitting (SNP) form appears to occur predictably and may reflect competition between two populations growing at different rates. Better survival of the slower-growing (DAP) form may explain its relatively easy retrieval from piliated SNP populations allowed to age on solid media. Pilin expression is associated with auto-agglutination in liquid suspension or broth cultures, and appears to be necessary but not sufficient for expression of the agar-pitting phenotype and for the formation of biofilms. Outer-membrane protein variation is seen in association with phase variation, but lipopolysaccharide expression is preserved in piliated as well as extensively passaged non-piliated isolates. The EagI/HhaI infrequent restriction site-PCR fingerprint, which has been previously used to discriminate between serotypes Marseille and Houston, is shown to alter with phase variation in vitro, and there is evidence that genetic change accompanies these events. The extent of genetic and phenotypic variability of phase-variant B. henselae has previously been underestimated. It may lead to new insights into the pathogenicity of this organism, and must be considered when interpreting data arising from such studies. PMID- 12634332 TI - Comparative assessment of virulence traits in Legionella spp. AB - Legionella pneumophila is a facultative intracellular pathogen that accounts for the majority of cases of Legionnaires' disease in the USA and Europe, but other Legionella spp. have been shown to cause disease. In contrast, Legionella longbeachae is the leading cause of Legionnaires' disease in Australia. The hallmark of Legionnaires' disease caused by L. pneumophila is the intracellular replication within phagocytes in the alveolar spaces, and the Dot/Icm type IV secretion system is essential for intracellular replication. Although it has been presumed that intracellular replication within phagocytes is the hallmark of other virulent legionellae, the virulence traits of Legionella spp. apart from L. pneumophila are not well defined. In this study, 27 strains of Legionella spp. belonging to 16 species that have been isolated from humans or from the environment were examined for five virulence traits exhibited by L. pneumophila: cytopathogenicity, intracellular replication within macrophages, induction of apoptosis/DNA fragmentation, pore-formation-mediated cytolysis of the host cell, and the presence of the dot/icm loci. The strains were divided into two broad groups (low and high cytopathogenic groups) based on cytopathogenicity assays using U937 human-derived macrophages. The other four virulence traits were evaluated in the low and high cytopathogenic groups of Legionella species. Most L. pneumophila serogroup 1 strains were highly cytopathogenic after 72 h, manifested high levels of intracellular growth, induced apoptosis/DNA fragmentation, and exhibited pore-forming activity. The majority of the other species were the low cytopathogenic group that did not induce apoptosis, neither did they exhibit pore-forming activity. All the species of legionellae tested have all the dot/icm loci, when examined by DNA hybridization. No correlation was found between cytopathogenicity and the other four pathogenic traits. PMID- 12634333 TI - Characterization of novel LPXTG-containing proteins of Staphylococcus aureus identified from genome sequences. AB - Twenty-one genes encoding surface proteins belonging to the LPXTG family have been identified by in silico analysis of six Staphylococcus aureus genome sequences. Eleven genes encode previously described proteins, while 10 have not yet been characterized. Of these, eight contain the cell-wall sorting signal LPXTG responsible for covalently anchoring proteins to the cell-wall peptidoglycan. The remaining two, SasF and SasD, harbour a single residue variation in the fourth position of the LPXTG motif (LPXAG). Western blotting of lysostaphin-solubilized S. aureus cell-wall proteins demonstrated the release of SasF in the cell-wall fraction, indicating that proteins carrying LPXAG are sorted normally. Analysis of primary sequences of the Staphylococcus aureus surface (Sas) proteins indicated that several share a similar structural organization and a common signal sequence with previously characterized LPXTG proteins of S. aureus and other Gram-positive cocci. Protein SasG has 128 residue B repeats that are almost identical at the DNA level. PCR analysis indicated that recombinants with repeat length variations are present in the bacterial population whereas they are not detectable in the B-repeat-encoding region of sdrD. The sasG and sasH genes are significantly associated with invasive disease isolates compared to nasal carriage isolates. Several IgG samples purified from patients recovering from S. aureus infections had higher titres against Sas proteins than control IgG, suggesting that expression occurred during infection in some patients. PMID- 12634334 TI - Production of enterolysin A by a raw milk enterococcal isolate exhibiting multiple virulence factors. AB - Even though enterococci are a common cause of human infection they can readily be isolated from a range of food sources, including various meat and dairy products. An enterococcal strain, DPC5280, which exhibits a broad spectrum of inhibition against many Gram-positive bacteria was recently isolated from an Irish raw milk sample. Characterization of the inhibition revealed that the strain exhibits haemolytic activity characteristic of the two-component lantibiotic cytolysin and also produces a heat-labile antimicrobial protein of 34 kDa. The latter protein displayed cell wall hydrolytic activity, as evidenced by zymogram gels containing autoclaved lactococcal cells. N-terminal sequencing of the purified protein yielded the sequence ASNEWS which is 100 % identical to enterolysin A (accession no. AF249740), a protein which shares 28 and 29 % identity to the Gly-Gly endopeptidases, lysostaphin and zoocin A, respectively. Indeed, amplification of entL from DPC5280 and sequencing revealed that the protein is 100 % identical to enterolysin A. The DPC5280 strain also contained the determinants associated with multiple virulence factors, including gelatinase, aggregation substance and multiple antibiotic resistance. The linkage of this cell-wall-degrading enzyme to other virulence factors in enterococci may contribute to the competitiveness of pathogenic enterococci when found in complex microbial environments such as food and the gastrointestinal tract. PMID- 12634335 TI - Catalase (KatA) and KatA-associated protein (KapA) are essential to persistent colonization in the Helicobacter pylori SS1 mouse model. AB - Helicobacter pylori infects the human gastric mucosa and elicits an aggressive inflammatory response. Despite the severity of the inflammatory response, the bacterium is able to persist and cause a chronic infection. It is believed that antioxidant defence mechanisms enable this organism to persist. Wild-type H. pylori strain SS1, and KatA- and KapA-deficient mutants, were used to infect C57/BL6 mice to test this hypothesis. Neither KatA nor KapA was essential for the initial colonization of H. pylori SS1 in the murine model of infection. The wild type SS1 colonized the gastric mucosa at significantly higher levels than both mutants throughout the 24-week experiment. Neither KatA- nor KapA-deficient mutants were able to maintain consistent ongoing colonization for the 24-week period, indicating the necessity of both KapA and KatA in sustaining a long-term infection. At 24 weeks, 5/10 mice inoculated with the KatA mutant and 2/10 mice inoculated with the KapA mutant were colonized, compared with 10/10 of the mice inoculated with the wild-type SS1. An increase in the severity of inflammation in the wild-type-inoculated mice appeared to correlate with the decline in colonization of animals inoculated with the mutants, suggesting that increased oxidative stress militated against continued infection by the mutants. These data indicate that KapA may be of equal or greater importance than KatA in terms of sustained infection on inflamed gastric mucosae. PMID- 12634336 TI - Regulation of the aerobic respiratory chain in the facultatively aerobic and hyperthermophilic archaeon Pyrobaculum oguniense. AB - The aerobic respiratory chain of Pyrobaculum oguniense is expressed constitutively even under anaerobic conditions. The membranes of both aerobically and anaerobically grown cells show oxygen consumption activity with NADH as substrate, bovine cytochrome c oxidase activity and TMPD oxidase activity. Spectroscopic analysis and haem analysis of membranes of aerobically grown cells show the presence of cytochrome b(559), cytochrome c(551) and haem Op1 containing cytochrome c oxidase in aerobically and anaerobically grown cells, and haem As containing cytochrome c oxidase in aerobically grown cells. The gene clusters of SoxB-type and SoxM-type haem copper oxidase and cytochrome bc complex have been cloned and sequenced and the regulation of these genes was analysed. The Northern blot analysis indicated that the constitutive transcription of the gene cluster of SoxB-type haem-copper oxidase and cytochrome bc complex is observed under both aerobic and anaerobic conditions, and the transcription of the operon of SoxM type haem-copper oxidase was stimulated under aerobic conditions. Furthermore, the presence of the binding residues for CuA in subunit II of both SoxB- and SoxM type haem-copper oxidase suggests that these haem-copper oxidases are cytochrome c oxidases. PMID- 12634337 TI - Construction and complementation of the first auxotrophic mutant in the spirochaete Leptospira meyeri. AB - In bacteria, the first reaction of the tryptophan biosynthetic pathway involves the conversion of chorismate and glutamine to anthranilate by the action of anthranilate synthase, which is composed of the alpha (trpE gene product) and beta (trpG gene product) subunits. In this study, the tryptophan biosynthetic gene trpE of the spirochaete Leptospira meyeri was interrupted by a kanamycin resistance cassette by homologous recombination. The trpE double cross-over mutant was not able to grow on solid or in liquid EMJH medium. In contrast, the trpE mutant showed a wild-type phenotype when tryptophan or anthranilate was added to the media, therefore showing that disruption of the L. meyeri trpE gene resulted in tryptophan auxotrophy. The authors have also characterized a second selectable marker that allows the construction of a spectinomycin-resistant L. meyeri-E. coli shuttle vector and the functional complementation of the L. meyeri trpE mutant. PMID- 12634338 TI - Characterization of AcmB, an N-acetylglucosaminidase autolysin from Lactococcus lactis. AB - A gene encoding a putative peptidoglycan hydrolase, named acmB, which is a paralogue of the major autolysin acmA gene, was identified in the Lactococcus lactis genome sequence. The acmB gene is transcribed in L. lactis MG1363 and its expression is modulated during cellular growth. The encoded AcmB protein has a modular structure with three domains: an N-terminal domain, especially rich in Ser, Thr, Pro and Asn residues, resembling a cell-wall-associated domain; a central domain homologous to the Enterococcus hirae muramidase catalytic domain; and a C-terminal domain of unknown function. A recombinant AcmB derivative, devoid of its N-terminal domain, was expressed in Escherichia coli. It exhibited hydrolysing activity on the peptidoglycan of several Gram-positive bacteria, including L. lactis. Though showing sequence similarity with enterococcal muramidase, AcmB has N-acetylglucosaminidase specificity. The acmB gene was inactivated in order to evaluate the role of the enzyme. AcmB does not appear to be involved in cell separation but contributes to cellular autolysis. PMID- 12634339 TI - Identification of the putrescine biosynthetic genes in Pseudomonas aeruginosa and characterization of agmatine deiminase and N-carbamoylputrescine amidohydrolase of the arginine decarboxylase pathway. AB - Putrescine can be synthesized either directly from ornithine by ornithine decarboxylase (ODC; the speC product) or indirectly from arginine via arginine decarboxylase (ADC; the speA product). The authors identified the speA and speC genes in Pseudomonas aeruginosa PAO1. The activities of the two decarboxylases were similar and each enzyme alone appeared to direct sufficient formation of the polyamine for normal growth. A mutant defective in both speA and speC was a putrescine auxotroph. In this strain, agmatine deiminase (the aguA product) and N carbamoylputrescine amidohydrolase (the aguB product), which were initially identified as the catabolic enzymes of agmatine, biosynthetically convert agmatine to putrescine in the ADC pathway: a double mutant of aguAB and speC was a putrescine auxotroph. AguA was purified as a homodimer of 43 kDa subunits and AguB as a homohexamer of 33 kDa subunits. AguA specifically deiminated agmatine with K(m) and K(cat) values of 0.6 mM and 4.2 s(-1), respectively. AguB was specific to N-carbamoylputrescine and the K(m) and K(cat) values of the enzyme for the substrate were 0.5 mM and 3.3 s(-1), respectively. Whereas AguA has no structural relationship to any known C-N hydrolases, AguB is a protein of the nitrilase family that performs thiol-assisted catalysis. Inhibition by SH reagents and the conserved cysteine residue in AguA and its homologues suggested that this enzyme is also involved in thiol-mediated catalysis. PMID- 12634341 TI - Correlation of the rate of protein synthesis and the third power of the RNA : protein ratio in Escherichia coli and Mycobacterium tuberculosis. AB - In order to further understand the different physiological states of the tubercle bacillus, a frame of reference was sought by first correlating the macromolecular compositions of Escherichia coli with specific growth rates and also with the rates of protein synthesis. Data for DNA : protein : RNA were converted to the average amounts of DNA [m(DNA(av))], protein [m(p(av))] and RNA [m(RNA(av))] per cell. The specific growth rate micro was found to be directly proportional to m(RNA(av))/m(p(av)). The specific protein synthesis rate per average cell [omega(p(av))] was shown to be directly proportional to the third power of the ratio m(RNA(av))/m(p(av)) which reflects the ribosome concentration. The equations derived were shown apply to both E. coli ( micro =1.73 h(-1)) and Mycobacterium bovis BCG ( micro =0.029 h(-1)). PMID- 12634340 TI - Autoinducer 2 activity in Escherichia coli culture supernatants can be actively reduced despite maintenance of an active synthase, LuxS. AB - Production of the signalling molecule (autoinducer-2) synthesized by LuxS has been proposed to be pivotal to a universal mechanism of inter-species bacterial cell-cell communication (quorum sensing); however recently the function of LuxS has been noted to be integral to central metabolism since it contributes to the activated methyl cycle. This paper shows that when Helicobacter pylori LuxS is overproduced in Escherichia coli, it forms cross-linkable multimers. These multimers persist at comparable levels after 24 h of growth if glucose is omitted from the growth medium; however, the levels of extracellular autoinducer-2 decline (Glucose Retention of AI-2 Levels: GRAIL). Glycerol, maltose, galactose, ribose and L-arabinose could substitute for glucose, but lactose, D-arabinose, acetate, citrate and pyruvate could not. Mutations in (i). metabolic pathways (glycolytic enzymes eno, pgk, pgm; galactose epimerase; the Pta-AckA pathway), (ii). sugar transport (pts components, rbs operon, mgl, trg), and (iii). regulators involved in conventional catabolic repression (crp, cya), cAMP independent catabolite repression (creC, fruR, rpoS,) the stringent response (relA, spoT) and the global carbon storage regulator (csrA) did not prevent GRAIL. Although the basis of GRAIL remains uncertain, it is clear that the mechanism is distinct from conventional catabolite repression. Moreover, GRAIL is not due to inactivation of the enzymic activity of LuxS, since in E. coli, LuxS contained within stationary-phase cells grown in the absence of glucose maintains its activity in vitro. PMID- 12634342 TI - Specificity of the interaction of RocR with the rocG-rocA intergenic region in Bacillus subtilis. AB - In Bacillus subtilis, expression of the rocG gene, encoding glutamate dehydrogenase, and the rocABC operon, involved in arginine catabolism, requires SigL (sigma(54))-containing RNA polymerase as well as RocR, a positive regulator of the NtrC/NifA family. The RocR protein was purified and shown to bind specifically to the intergenic region located between rocG and the rocABC operon. DNaseI footprinting experiments were used to define the RocR-binding site as an 8 bp inverted repeat, separated by one base pair, forming an imperfect palindrome which is present twice within the rocG-rocABC intergenic region, acting as both a downstream activating sequence (DAS) and an upstream activating sequence (UAS). Point mutations in either of these two sequences significantly lowered expression of both rocG and rocABC. This bidirectional enhancer element retained partial activity even when moved 9 kb downstream of the rocA promoter. Electron microscopy experiments indicated that an intrinsically curved region is located between the UAS/DAS region and the promoter of the rocABC operon. This curvature could facilitate interaction of RocR with sigma(54)-RNA polymerase at the rocABC promoter. PMID- 12634343 TI - Expression of the glycolytic gapA operon in Bacillus subtilis: differential syntheses of proteins encoded by the operon. AB - Glycolysis is one of the central routes of carbon catabolism in Bacillus subtilis. Several glycolytic enzymes, including the key enzyme glyceraldehyde-3 phosphate dehydrogenase, are encoded in the hexacistronic gapA operon. Expression of this operon is induced by a variety of sugars and amino acids. Under non inducing conditions, expression is repressed by the CggR repressor protein, the product of the promoter-proximal gene of the operon. Here, it is shown that the amount of glyceraldehyde-3-phosphate dehydrogenase encoded by the second gene of the operon exceeds that of the CggR repressor by about 100-fold. This differential synthesis was attributed to an mRNA processing event that takes place at the 3' end of the cggR open reading frame and to differential segmental stabilities of the resulting cleavage products. The mRNA specifying the truncated cggR gene is quickly degraded, whereas the downstream processing products encompassing gapA are quite stable. This increased stability is conferred by the presence of a stem-loop structure at the 5' end of the processed mRNAs. Mutations were introduced in the region of the cleavage site. A mutation affecting the stability of the stem-loop structure immediately downstream of the processing site had two effects. First, the steady-state transcript pattern was drastically shifted towards the primary transcripts; second, the stability of the processed mRNA containing the destabilized stem-loop structure was strongly decreased. This results in a reduction of the amount of glyceraldehyde-3-phosphate dehydrogenase in the cell. It is concluded that mRNA processing is involved in differential syntheses of the proteins encoded by the gapA operon. PMID- 12634344 TI - Studies on the Escherichia coli glucose-specific permease, PtsG, with a point mutation in its N-terminal amphipathic leader sequence. AB - Previous work has resulted in the isolation of several mutant glucose permeases (II(Glc) or PtsG) of the Escherichia coli phosphotransferase system (PTS) with altered N-terminal amphipathic leader sequences. The mutations were reported to (1). broaden permease substrate specificity, (2). promote facilitated diffusion of some sugars and (3). increase ptsG gene transcription. Detailed biochemical analyses were conducted, showing that one such mutant (V12F-II(Glc)) (1). contains dramatically increased amounts of II(Glc), (2). displays correspondingly increased in vitro phosphorylation and in vivo transport activities, (3). shows increased utilization of several metabolizable sugars and (4). shows decreased susceptibility to detergent activation. These results are interpreted as suggesting that the V12F substitution in the N-terminal amphipathic leader sequence of II(Glc) alters the facility with which the permease is integrated into the membrane. Consequent changes in conformation alter its catalytic properties and increase its affinity for the pleiotropic transcriptional repressor, Mlc. These changes together are proposed to promote transcription of the ptsG gene and account for the observed phenotypic changes. PMID- 12634345 TI - Transcription analysis of the dnaA gene and oriC region of the chromosome of Mycobacterium smegmatis and Mycobacterium bovis BCG, and its regulation by the DnaA protein. AB - The regions flanking the Mycobacterium dnaA gene have extensive sequence conservation, and comprise various DnaA boxes. Comparative analysis of the dnaA promoter and oriC region from several mycobacterial species revealed that the localization, spacing and orientation of the DnaA boxes are conserved. Detailed transcriptional analysis in M. smegmatis and M. bovis BCG shows that the dnaN gene of both species and the dnaA gene of M. bovis BCG are transcribed from two promoters, whereas the dnaA gene of M. smegmatis is transcribed from a single promoter. RT-PCR with total RNA showed that dnaA and dnaN were expressed in both species at all growth stages. Analysis of the promoter activity using dnaA-gfp fusion plasmids and DnaA expression plasmids indicates that the dnaA gene is autoregulated, although the degree of transcriptional autorepression was moderate. Transcription was also detected in the vicinity of oriC of M. bovis BCG, but not of M. smegmatis. These results suggest that a more complex transcriptional mechanism may be involved in the slow-growing mycobacteria, which regulates the expression of dnaA and initiation of chromosomal DNA replication. PMID- 12634346 TI - Stable low-copy-number Staphylococcus aureus shuttle vectors. AB - A series of Staphylococcus aureus-Escherichia coli shuttle vectors were constructed which contained the replication and maintenance functions of the S. aureus theta-mode multiresistance plasmid pSK1. The utility of the newly constructed vectors was demonstrated by the successful cloning and expression of several genes that had previously proven difficult to express in S. aureus. Additional vectors which permit the generation of transcriptional and translational fusions to an S. aureus blaZ reporter gene were also produced and subsequently employed to determine the relative strengths in S. aureus of a number of promoters. By utilizing the theta-mode replication functions of pSK1, the shuttle vectors described largely avoided the segregational and structural stability problems frequently encountered with Gram-positive rolling-circle-based vectors. In addition, these plasmids represent vectors which are suitable for the analysis of genes in S. aureus at low copy number. PMID- 12634347 TI - Expansion of growth substrate range in Pseudomonas putida F1 by mutations in both cymR and todS, which recruit a ring-fission hydrolase CmtE and induce the tod catabolic operon, respectively. AB - Pseudomonas putida F1 can assimilate benzene, toluene and ethylbenzene using the toluene degradation pathway, and can also utilize p-cymene via p-cumate using the p-cymene and p-cumate catabolic pathways. In the present study, P. putida F1 strains were isolated that were adapted to assimilate new substrates such as n propylbenzene, n-butylbenzene, cumene and biphenyl, and the molecular mechanisms of genetic adaptation to an expanded range of aromatic hydrocarbons were determined. Nucleotide sequence analyses showed that the selected strains have mutations in the cymR gene but not in todF gene. The impairment of the repressor CymR by mutation led to the constitutive expression of CmtE, a meta-cleavage product hydrolase from the cmt operon. This study also showed that CmtE has a broad range of substrates and can hydrolyse meta-cleavage products formed from biphenyl and other new growth substrates via the toluene degradation pathway. However, the artificially constructed strain P. putida F1(cymR : : Tc(r)) and a recombinant P. putida F1, which expressed CmtE constitutively, could not grow on the new substrates. The adapted strains possess the tod operon, which is induced by new growth substrates that are poor inducers of wild-type P. putida F1. When the todS gene from the adapted strains was introduced in a trans manner to P. putida F1(cymR : : Tc(r)), the resulting recombinant strains were able to grow on biphenyl and other new substrates. This finding indicates that the TodS sensor was altered to recognize these substrates and this conclusion was confirmed by nucleotide sequence analyses. Amino acid substitutions were found in the regions corresponding to the receiver domain and the second PAS domain and their boundaries in the TodS protein. These results showed that P. putida F1 adapted strains capable of growth on n-propylbenzene, n-butylbenzene, cumene and biphenyl possess mutations to employ CmtE and to induce the tod catabolic operon by the new growth substrates. PMID- 12634348 TI - 16S-23S rRNA intergenic spacer region sequence variation in Streptococcus thermophilus and related dairy streptococci and development of a multiplex ITS SSCP analysis for their identification. AB - The 16S-23S rRNA internal transcribed spacer (ITS) region of several Streptococcus thermophilus strains and some related dairy streptococci, S. macedonicus, S. salivarius and S. bovis, was analysed by sequence analysis. All the Streptococcus species were easily discriminated on the basis of sequence variations principally located upstream and downstream of the region encompassing the double-stranded processing sites and the tRNA(Ala) gene. Comparison between tRNA(Ala) gene sequences highlighted a high level of sequence conservation among the Streptococcus species investigated despite their belonging to separated phylogenetic clusters, i.e. the S. salivarius and S. bovis rRNA groups. A low but significant degree of variability was detected among the S. thermophilus strains, allowing the identification of four different ITS sequences. Similarity analysis of the ITS sequences showed that the Streptococcus species were clustered in two main branches, one containing S. macedonicus and S. bovis strains, and one containing S. thermophilus and S. salivarius strains. With the aim of developing a rapid tool for the identification of the dairy streptococci species a multiplex ITS-SSCP analysis of two discrete regions within the ITS locus was carried out. PMID- 12634349 TI - Molecular clocks and the puzzle of RNA virus origins. PMID- 12634351 TI - Alpha4beta1 integrin acts as a cell receptor for murine polyomavirus at the postattachment level. AB - The initial interaction of murine polyomavirus (Py) with host cells occurs through direct binding of the major capsid protein VP1 with cell membrane molecules containing terminal sialic acids; however, these Py receptor molecules have not yet been identified. Analysis of the capsid protein primary sequences of all murine strains revealed the presence of integrin ligand motifs in the DE and EF loops of VP1 (LDV and DLXXL, respectively) and at the N terminus of VP2 (DGE). We show that infectivity of the Py A2 strain in mouse Swiss 3T3 fibroblasts is significantly reduced only in the presence of natural integrin ligands carrying an LDV motif or antibodies directed against the alpha4 and beta1 integrin subunits. Furthermore, we demonstrate that expression of the alpha4 subunit in the alpha4-deficient BALB/c 3T3 cells increases viral infectivity. Addition of alpha4 function-blocking antibodies, prior to or after virus adsorption, blocks this increased infectivity without affecting virus binding to cells. Taken together, these data indicate that expression of alpha4 integrin enhances permissivity to Py, probably by acting as one of the postattachment receptors. PMID- 12634352 TI - Evidence that binding of cucumber necrosis virus to vector zoospores involves recognition of oligosaccharides. AB - Despite the importance of vectors in natural dissemination of plant viruses, relatively little is known about the molecular features of viruses and vectors that permit their interaction in nature. Cucumber necrosis virus (CNV) is a small spherical virus whose transmission in nature is facilitated by zoospores of the fungus Olpidium bornovanus. Previous studies have shown that specific regions of the CNV capsid are involved in transmission and that transmission defects in several CNV transmission mutants are due to inefficient attachment of virions to the zoospore surface. In this study, we have undertaken to determine if zoospores contain specific receptors for CNV. We show that in vitro binding of CNV to zoospores is saturable and that vector zoospores bind CNV more efficiently than nonvector zoospores. Further studies show that treatment of zoospores with periodate and trypsin reduces CNV binding, suggesting the involvement of glycoproteins in zoospore attachment. In virus overlay assays, CNV binds to several proteins, whereas CNV transmission mutants either fail to bind or bind at significantly reduced levels. The possible involvement of specific sugars in attachment was investigated by incubating CNV with zoospores in the presence of various sugars. Two mannose derivatives (methyl alpha-D-mannopyranoside and D mannosamine), as well as three mannose-containing oligosaccharides (mannotriose, alpha3,alpha6-mannopentaose, and yeast mannan) and L-(-)-fucose, all inhibited CNV binding at relatively low concentrations. Taken together, our studies suggest that binding of CNV to zoospores is mediated by specific mannose and/or fucose containing oligosaccharides. This is the first time sugars have been implicated in transmission of a plant virus. PMID- 12634350 TI - Alpha interferon induces distinct translational control programs to suppress hepatitis C virus RNA replication. AB - Hepatitis C virus (HCV) infection is treated with interferon (IFN)-based therapy. The mechanisms by which IFN suppresses HCV replication are not known, and only limited efficacy is achieved with therapy because the virus directs mechanisms to resist the host IFN response. In the present study we characterized the effects of IFN action upon the replication of two distinct quasispecies of an HCV replicon whose encoded NS5A protein exhibited differential abilities to bind and inhibit protein kinase R (PKR). Metabolic labeling experiments revealed that IFN had little overall effect upon HCV protein stability or polyprotein processing but specifically blocked translation of the HCV RNA, such that the replication of both viral quasispecies was suppressed by IFN treatment of the Huh7 host cells. However, within cells expressing an NS5A variant that inhibited PKR, we observed a reduced level of eukaryotic initiation factor 2 alpha subunit (eIF2alpha) phosphorylation and a concomitant increase in HCV protein synthetic rates, enhancement of viral RNA replication, and a partial rescue of viral internal ribosome entry site (IRES) function from IFN suppression. Assessment of the ribosome distribution of the HCV replicon RNA demonstrated that the NS5A-mediated block in eIF2alpha phosphorylation resulted in enhanced recruitment of the HCV RNA into polyribosome complexes in vivo but only partially rescued the RNA from polyribosome dissociation induced by IFN treatment. Examination of cellular proteins associated with HCV-translation complexes in IFN-treated cells identified the P56 protein as an eIF3-associated factor that fractionated with the initiator ribosome-HCV RNA complex. Importantly, we found that P56 could independently suppress HCV IRES function both in vitro and in vivo, but a mutant P56 that was unable to bind eIF3 had no suppressive action. We conclude that IFN blocks HCV replication through translational control programs involving PKR and P56 to, respectively, target eIF2- and eIF3-dependent steps in the viral RNA translation initiation process. PMID- 12634353 TI - American genotype structures decrease dengue virus output from human monocytes and dendritic cells. AB - The dengue virus type 2 structures probably involved in human virulence were previously defined by sequencing the complete genome of both American and Southeast (SE) Asian genotype templates in patient serum (K. C. Leitmeyer et al., J. Virol. 73:4738-4747, 1999). We have now evaluated the effects of introducing a mutation in the envelope glycoprotein (E) gene and/or replacement of 5'- and 3' nontranslated regions on dengue virus replication in human primary cell cultures. A series of chimeric infectious clones were generated containing different combinations of American and SE Asian genotype sequences. Some of the chimeric viruses had altered plaque morphology in mammalian cells; however, they replicated at similar rates in mosquito cells as measured by quantitative reverse transcription-PCR and plaque assay. Although susceptibility to virus infection varied from donor to donor in experiments using human macrophage and dendritic cells, we were able to measure consistent differences in viral RNA output per infected cell. Using this measurement, we demonstrated that the chimeric virus containing the E mutation had a lower virus output compared to the parental infectious clone. A larger reduction in virus output was observed for the triple mutant and the wild-type, American genotype virus from which chimeric inserts were derived. It appears that the three changes function synergistically, although the E mutation alone gives a lower output compared to the 5'- and 3' terminal mutations. The data suggest that these changes may be responsible for decreased dengue virus replication in human target cells and for virulence characteristics during infection. PMID- 12634354 TI - The leader of human immunodeficiency virus type 1 genomic RNA harbors an internal ribosome entry segment that is active during the G2/M phase of the cell cycle. AB - The 5' leader of the human immunodeficiency virus type 1 (HIV-1) genomic RNA contains highly structured domains involved in key steps of the viral life cycle. These RNA domains inhibit cap-dependent protein synthesis. Here we report that the HIV-1 5' leader harbors an internal ribosome entry site (IRES) capable of driving protein synthesis during the G(2)/M cell cycle phase in which cap dependent initiation is inhibited. The HIV-1 IRES was delineated with bicistronic mRNAs in in vitro and ex vivo assays. The HIV-1 leader IRES spans nucleotides 104 to 336 and partially overlaps the major determinants of genomic RNA packaging. These data strongly suggest that, as for HIV-1 transcription, IRES-mediated translation initiation could play an important role in virus replication during virus-induced G(2)/M cell cycle arrest. PMID- 12634355 TI - Hepatitis C virus NS3 ATPases/helicases from different genotypes exhibit variations in enzymatic properties. AB - The NS3 ATPase/helicase was isolated and characterized from three different infectious clones of hepatitis C virus (HCV). One helicase was from a genotype that normally responds to therapy (Hel-2a), and the other two were from more resistant genotypes, 1a (Hel-1a) and 1b (Hel-1b). Although the differences among these helicases are generally minor, all three enzymes have distinct properties. Hel-1a is less selective for nucleoside triphosphates, Hel-1b hydrolyzes nucleoside triphosphates less rapidly, and Hel-2a unwinds DNA more rapidly and binds DNA more tightly than the other two enzymes. Unlike related proteins, different nucleic acid sequences stimulate ATP hydrolysis by HCV helicase at different maximum rates and with different apparent efficiencies. This nucleic acid stimulation profile is conserved among the enzymes, but it does not result entirely from differential DNA-binding affinities. Although the amino acid sequences of the three proteins differ by up to 15%, one variant amino acid that is critical for helicase action was identified. NS3 residue 450 is a threonine in Hel-1a and Hel-1b and is an isoleucine in Hel-2a. A mutant Hel-1a with an isoleucine substituted for threonine 450 unwinds DNA more rapidly and binds DNA more tightly than the parent protein. PMID- 12634357 TI - Rapid localization of Gag/GagPol complexes to detergent-resistant membrane during the assembly of human immunodeficiency virus type 1. AB - During human immunodeficiency virus type 1 (HIV-1) assembly in HIV-1-transfected COS7 cells, almost all steady-state Gag/Gag and Gag/GagPol complexes are membrane bound. However, exposure to 1% Triton X-100 gives results indicating that while all Gag/GagPol complexes remain associated with the detergent-resistant membrane (DRM), only 30% of Gag/Gag complexes are associated with the DRM. Analysis of the localization of newly synthesized Gag/Gag and Gag/GagPol to the membrane indicates that after a 10-min pulse with radioactive [(35)S]Cys-[(35)S]Met, all newly synthesized Gag/GagPol is found at the DRM. Only 30% of newly synthesized Gag/Gag moves to the membrane, and at 0 min of chase, only 38% of this membrane bound Gag/Gag is associated with the DRM. During the first 30 min of chase, most membrane-bound Gag/Gag moves to the DRM, while between 30 and 60 min of chase, there is a significant decrease in membrane-bound Gag/Gag and Gag/GagPol. Since the localization of newly synthesized Gag/Gag to the DRM and the interaction of GagPol with Gag both depend upon Gag multimerization, the rapid localization of GagPol to the DRM probably reflects the interaction of all newly synthesized GagPol with the first newly synthesized polymeric Gag to associate with the DRM. PMID- 12634356 TI - Human immunodeficiency virus type 1 (HIV-1) Vpr enhances expression from unintegrated HIV-1 DNA. AB - Retroviral DNA synthesized prior to integration, termed unintegrated viral DNA, is classically believed to be transcriptionally inert and to serve only as a precursor to the transcriptionally active integrated proviral DNA form. However, it has recently been found to be expressed under some circumstances during human immunodeficiency virus type 1 (HIV-1) replication and may play a significant role in HIV-1 pathogenesis. HIV-1 Vpr is a virion-associated accessory protein that is critical for HIV-1 replication in nondividing cells and induces cell cycle arrest and apoptosis. We find that Vpr, either expressed de novo or released from virions following viral entry, is essential for unintegrated viral DNA expression. HIV-1 mutants defective for integration in either the integrase catalytic domain or the cis-acting att sites can express unintegrated viral DNA at levels similar to that of wild-type HIV-1, but only in the presence of Vpr. In the absence of Vpr, the expression of unintegrated viral DNA decreases 10- to 20 fold. Vpr does not affect the efficiency of integration from integrase-defective HIV-1. Vpr-mediated enhancement of expression from integrase-defective HIV-1 requires that the viral DNA be generated in cells through infection and is mediated via a template that declines over time. Vpr activation of expression does not require exclusive nuclear localization of Vpr nor does it correlate with Vpr-mediated cell cycle arrest. These results attribute a new function to HIV-1 Vpr and implicate Vpr as a critical component in expression from unintegrated HIV 1 DNA. PMID- 12634358 TI - Organization of the vesicular stomatitis virus glycoprotein into membrane microdomains occurs independently of intracellular viral components. AB - The glycoprotein (G protein) of vesicular stomatitis virus (VSV) is primarily organized in plasma membranes of infected cells into membrane microdomains with diameters of 100 to 150 nm, with smaller amounts organized into microdomains of larger sizes. This organization has been observed in areas of the infected-cell plasma membrane that are outside of virus budding sites as well as in the envelopes of budding virions. These observations raise the question of whether the intracellular virion components play a role in organizing the G protein into membrane microdomains. Immunogold-labeling electron microscopy was used to analyze the distribution of the G protein in arbitrarily chosen areas of plasma membranes of transfected cells that expressed the G protein in the absence of other viral components. Similar to the results with virus-infected cells, the G protein was organized predominantly into membrane microdomains with diameters of approximately 100 to 150 nm. These results indicate that internal virion components are not required to concentrate the G protein into membrane microdomains with a density similar to that of virus envelopes. To determine if interactions between the G protein cytoplasmic domain and internal virion components were required to create a virus budding site, cells infected with recombinant VSVs encoding truncation mutations of the G protein cytoplasmic domain were analyzed by immunogold-labeling electron microscopy. Deletion of the cytoplasmic domain of the G protein did not alter its partitioning into the 100- to 150-nm microdomains, nor did it affect the incorporation of the G protein into virus envelopes. These data support a model for virus assembly in which the G protein has the inherent property of partitioning into membrane microdomains that then serve as the sites of assembly of internal virion components. PMID- 12634359 TI - Distinct mechanisms of neutralization by monoclonal antibodies specific for sites in the N-terminal or C-terminal domain of murine leukemia virus SU. AB - The epitope specificities and functional activities of monoclonal antibodies (MAbs) specific for the murine leukemia virus (MuLV) SU envelope protein subunit were determined. Neutralizing antibodies were directed towards two distinct sites in MuLV SU: one overlapping the major receptor-binding pocket in the N-terminal domain and the other involving a region that includes the most C-terminal disulfide-bonded loop. Two other groups of MAbs, reactive with distinct sites in the N-terminal domain or in the proline-rich region (PRR), did not neutralize MuLV infectivity. Only the neutralizing MAbs specific for the receptor-binding pocket were able to block binding of purified SU and MuLV virions to cells expressing the ecotropic MuLV receptor, mCAT-1. Whereas the neutralizing MAbs specific for the C-terminal domain did not interfere with the SU-mCAT-1 interaction, they efficiently inhibited cell-to-cell fusion mediated by MuLV Env, indicating that they interfered with a postattachment event necessary for fusion. The C-terminal domain MAbs displayed the highest neutralization titers and binding activities. However, the nonneutralizing PRR-specific MAbs bound to intact virions with affinities similar to those of the neutralizing receptor binding pocket-specific MAbs, indicating that epitope exposure, while necessary, is not sufficient for viral neutralization by MAbs. These results identify two separate neutralization domains in MuLV SU and suggest a role for the C-terminal domain in a postattachment step necessary for viral fusion. PMID- 12634360 TI - CC chemokine ligand 3 (CCL3) regulates CD8(+)-T-cell effector function and migration following viral infection. AB - Chemokines induce the directional migration of targeted populations of leukocytes during periods of inflammation. Moreover, these molecules also regulate T-cell activation and differentiation following antigenic stimulation. In the present study, the contributions of the CC chemokine ligand 3 (CCL3) to the differentiation and migration of effector T cells in response to viral infection of the central nervous system (CNS) were analyzed. CCL3(-/-) mice infected with mouse hepatitis virus exhibited a significant reduction of virus-specific CD8(+) T cells within the CNS, correlating with delayed viral clearance. Decreased infiltration of CD8(+) T cells into infected CCL3(-/-) mice was associated with enhanced accumulation of primed CD8(+) T cells in cervical lymph nodes. Although virus-specific CD8(+) T cells from CCL3(-/-) mice were CD44(high), they remained CD62L(high) and CD25(low), retained CCR7 expression, and contained limited transcripts of the proinflammatory chemokine receptors CCR5 and CXCR3 compared with virus-specific CD8(+) T cells from CCL3(+/+) mice. Furthermore, the absence of CCL3 impaired the cytokine production and cytolytic activity of CD8(+) T cells. In addition, macrophage accumulation within the CNS was significantly decreased in infected CCL3(-/-) mice, correlating with reduced demyelination. These results suggest that CCL3 not only mediates macrophage chemotaxis but also significantly enhances differentiation of primed CD8(+) T cells into effector cells and their release into circulation, thus potentiating effective migration to the site of infection. PMID- 12634361 TI - Unusual properties of adenovirus E2E transcription by RNA polymerase III. AB - In adenovirus type 5-infected cells, RNA polymerase III transcription of a gene superimposed on the 5' end of the E2E RNA polymerase II transcription unit produces two small (<100-nucleotide) RNAs that accumulate to low steady-state concentrations (W. Huang, R. Pruzan, and S. J. Flint, Proc. Natl. Acad. Sci. USA 91:1265-1269, 1984). To gain a better understanding of the function of this RNA polymerase III transcription, we have examined the properties of the small E2E RNAs and E2E RNA polymerase III transcription in more detail. The accumulation of cytoplasmic E2E RNAs and the rates of E2E transcription by the two RNA polymerases during the infectious cycle were analyzed by using RNase T(1) protection and run-on transcription assays, respectively. Although the RNA polymerase III transcripts were present at significantly lower concentrations than E2E mRNA throughout the period examined, E2E transcription by RNA polymerase III was found to be at least as efficient as that by RNA polymerase II. The short half-lifes of the small E2E RNAs estimated by using the actinomycin D chase method appear to account for their limited accumulation. The transcription of E2E sequences by RNA polymerase II and III in cells infected by recombinant adenoviruses carrying ectopic E2E-CAT (chloramphenicol transferase) reporter genes with mutations in E2E promoter sequences was also examined. The results of these experiments indicate that recognition of the E2E promoter by the RNA polymerase II transcriptional machinery in infected cells limits transcription by RNA polymerase III, and vice versa. Such transcriptional competition and the properties of E2E RNAs made by RNA polymerase III suggest that the function of this viral RNA polymerase III transcription unit is unusual. PMID- 12634362 TI - Transcriptional suppression of in vitro-integrated human immunodeficiency virus type 1 does not correlate with proviral DNA methylation. AB - Persistence of human immunodeficiency virus type 1 (HIV-1) constitutes a major obstacle in the control of HIV-1 infection. Here we investigated whether the CpG methylation of the HIV-1 promoter can directly influence the expression of the HIV-1 genome and thereby contribute to the persistence and latency of HIV-1. The levels of CpG methylation in the promoter of HIV-1 were studied after bisulfite induced modification of DNA in five Jurkat clonal cell lines transduced by an HIV 1 long terminal repeat (LTR)-driven retroviral vector and expressing enhanced green fluorescent protein (GFP) and in primary resting memory T cells challenged with HIV-1 or with an HIV-1-derived retroviral vector. Basal HIV-1 promoter activities were low or undetectable in three tested HIV-1 LTR-GFP clones, one of which encoded the Tat protein, and they reached medium or high levels in two other clones. The CpG dinucleotide that occurred in a latently infected clonal cell line 240 nucleotides upstream of the transcription start remained methylated after reactivation of HIV-1 transcription with 10 nM phorbol-12-myristate-13 acetate. In two clones showing a medium promoter activity and in resting memory T cells, the HIV-1 LTR was generally not methylated. Our results show that the methylation profiles of the HIV-1 LTR, including those present in latently infected cells, are low and do not correlate with the transcriptional activity. We suggest that, in a noncloned cellular population in which the HIV-1 proviruses are randomly integrated in the human genome, HIV-1 latency is imperfectly controlled by CpG methylation and is inherently accompanied by residual replication. PMID- 12634363 TI - Tumor necrosis factor alpha inhibition of hepatitis B virus replication involves disruption of capsid Integrity through activation of NF-kappaB. AB - Chronic infection by hepatitis B virus results from an inability to clear the virus, which is associated with liver disease and liver cancer. Tumor necrosis factor alpha (TNF-alpha) is associated with noncytopathic clearance of hepatitis B virus in animal models. Here we demonstrate that the nuclear factor kappaB (NF kappaB) signaling pathway is a central mediator of inhibition of hepatitis B virus by TNF-alpha and we describe the molecular mechanism. TNF-alpha is shown to suppress hepatitis B virus DNA replication without cell killing by disrupting the formation or stability of cytoplasmic viral capsids through a pathway requiring the NF-kappaB-activating inhibitor of kappaB kinase complex IKK-alpha/beta and active transcription factor NF-kappaB. Hepatitis B virus replication could also be inhibited and viral capsid formation could be disrupted in the absence of TNF alpha solely by overexpression of IKK-alpha/beta or strong activation of NF kappaB. In contrast, inhibition of NF-kappaB signaling stimulated viral replication, demonstrating that HBV replication is both positively and negatively regulated by the level of activity of the NF-kappaB pathway. Studies are presented that exclude the possibility that HBV inhibition by NF-kappaB is carried out by secondary production of gamma interferon or alpha/beta interferon. These results identify a novel mechanism for noncytopathic suppression of hepatitis B virus replication that is mediated by the NF-kappaB signaling pathway and activated by TNF-alpha. PMID- 12634364 TI - Mutation of single hydrophobic residue I27, L35, F39, L58, L65, L67, or L71 in the N terminus of VP5 abolishes interaction with the scaffold protein and prevents closure of herpes simplex virus type 1 capsid shells. AB - Protein-protein interactions drive the assembly of the herpes simplex virus type 1 (HSV-1) capsid. A key interaction occurs between the C-terminal tail of the scaffold protein (pre-22a) and the major capsid protein (VP5). Previously (Z. Hong, M. Beaudet-Miller, J. Durkin, R. Zhang, and A. D. Kwong, J. Virol. 70:533 540, 1996) it was shown that the minimal domain in the scaffold protein necessary for this interaction was composed of a hydrophobic amphipathic helix. The goal of this study was to identify the hydrophobic residues in VP5 important for this bimolecular interaction. Results from the genetic analysis of second-site revertant virus mutants identified the importance of the N terminus of VP5 for the interaction with the scaffold protein. This allowed us to focus our efforts on a small region of this large polypeptide. Twenty-four hydrophobic residues, starting at L23 and ending at F84, were mutated to alanine. All the mutants were first screened for interaction with pre-22a in the yeast two-hybrid assay. From this in vitro assay, seven residues, I27, L35, F39, L58, L65, L67, and L71, that eliminated the interaction when mutated were identified. All 24 mutants were introduced into the virus genome with a genetic marker rescue/marker transfer system. For this system, viruses and cell lines that greatly facilitated the introduction of the mutants into the genome were made. The same seven mutants that abolished interaction of VP5 with pre-22a resulted in an absolute requirement for wild-type VP5 for growth of the viruses. The viruses encoding these mutations in VP5 were capable of forming capsid shells comprised of VP5, VP19C, VP23, and VP26, but the closure of these shells into an icosahedral structure was prevented. Mutation at L75 did not affect the ability of this protein to interact with pre-22a, as judged from the in vitro assay, but this mutation specified a lethal effect for virus growth and abolished the formation of any detectable assembled structure. Thus, it appears that the L75 residue is important for another essential interaction of VP5 with the capsid shell proteins. The congruence of the data from the previous and present studies demonstrates the key roles of two regions in the N terminus of this large protein that are crucial for this bimolecular interaction. Thus, residues I27, L35, and F39 comprise the first subdomain and residues L58, L65, L67 and L71 comprise a second subdomain of VP5. These seven hydrophobic residues are important for the interaction of VP5 with the scaffold protein and consequently the formation of an icosahedral shell structure that encloses the viral genome. PMID- 12634365 TI - Possible role of dimerization in human immunodeficiency virus type 1 genome RNA packaging. AB - The dimer initiation site/dimer linkage sequence (DIS/DLS) region in the human immunodeficiency virus type 1 (HIV-1) RNA genome is suggested to play important roles in various steps of the virus life cycle. However, due to the presence of a putative DIS/DLS region located within the encapsidation signal region (E/psi), it is difficult to perform a mutational analysis of DIS/DLS without affecting the packaging of RNA into virions. Recently, we demonstrated that duplication of the DIS/DLS region in viral RNA caused the production of partially monomeric RNAs in virions, indicating that the region indeed mediated RNA-RNA interaction. We utilized this system to assess the precise location of DIS/DLS in the 5' region of the HIV-1 genome with minimum effect on RNA packaging. We found that the entire lower stem of the U5/L stem-loop was required for packaging, whereas the region important for dimer formation was only 10 bases long within the lower stem of the U5/L stem-loop. The R/U5 stem-loop was required for RNA packaging but was completely dispensable for dimer formation. The SL1 lower stem was important for both dimerization and packaging, but surprisingly, deletion of the palindromic sequence at the top of the loop only partially affected dimerization. These results clearly indicated that the E/psi of HIV-1 is much larger than the DIS/DLS and that the primary DIS/DLS is completely included in the E/psi. Therefore, it is suggested that RNA dimerization is a part of RNA packaging, which requires multiple steps. PMID- 12634367 TI - Fcgamma receptor-mediated suppression of human immunodeficiency virus type 1 replication in primary human macrophages. AB - Permissiveness of monocytes and macrophages to human immunodeficiency virus (HIV) infection is modulated by various stimuli. In this study we demonstrate that stimulation of primary monocytes and monocyte-derived macrophages (MDM) through the receptors for the Fc portion of immunoglobulin G (IgG) (FcgammaR) inhibits HIV type 1 (HIV-1) replication. Viral p24 production was decreased by 1.5 to 3 log units in MDM infected with both R5 and X4 HIV-1 strains upon stimulation by immobilized IgG but not upon stimulation by soluble IgG or by F(ab')(2) IgG fragments. Although MDM activation by immobilized IgG induced high levels of macrophage-derived chemokine secretion as well as a sustained down-regulation of CD4 and a transient decrease in CCR5 expression, these factors did not appear to play a major role in the suppression of HIV-1 replication. Single-cycle infection of FcgammaR-stimulated MDM with HIV-1 virions pseudotyped with either HIV-1 R5 or vesicular stomatitis virus G envelopes was inhibited, suggesting a postentry restriction of viral replication. PCR analyses of HIV-1 DNA intermediate replication forms suggested that reverse transcription is not affected by stimulation with immobilized human IgG, at least during the first replication cycle. The accumulation of PCR products corresponding to nuclear unintegrated two long-terminal-repeat circles and the relative decrease of integrated HIV-1 DNA signals suggest an inhibition of proviral integration. Our data, showing that FcgammaR-mediated activation of MDM is a potent mechanism of HIV-1 suppression, raise the possibility that FcgammaR cross-linking by immune complexes may contribute to the control of viral replication in macrophages. PMID- 12634366 TI - Hepatitis C virus glycoproteins interact with DC-SIGN and DC-SIGNR. AB - DC-SIGN and DC-SIGNR are two closely related membrane-associated C-type lectins that bind human immunodeficiency virus (HIV) envelope glycoprotein with high affinity. Binding of HIV to cells expressing DC-SIGN or DC-SIGNR can enhance the efficiency of infection of cells coexpressing the specific HIV receptors. DC-SIGN is expressed on some dendritic cells, while DC-SIGNR is localized to certain endothelial cell populations, including hepatic sinusoidal endothelial cells. We found that soluble versions of the hepatitis C virus (HCV) E2 glycoprotein and retrovirus pseudotypes expressing chimeric forms of both HCV E1 and E2 glycoproteins bound efficiently to DC-SIGN and DC-SIGNR expressed on cell lines and primary human endothelial cells but not to other C-type lectins tested. Soluble E2 bound to immature and mature human monocyte-derived dendritic cells (MDDCs). Binding of E2 to immature MDDCs was dependent on DC-SIGN interactions, while binding to mature MDDCs was partly independent of DC-SIGN, suggesting that other cell surface molecules may mediate HCV glycoprotein interactions. HCV interactions with DC-SIGN and DC-SIGNR may contribute to the establishment or persistence of infection both by the capture and delivery of virus to the liver and by modulating dendritic cell function. PMID- 12634369 TI - Influenza A virus-infected hosts boost an invasive type of Streptococcus pyogenes infection in mice. AB - The apparent worldwide resurgence of invasive Streptococcus pyogenes infection in the last two decades remains unexplained. At present, animal models in which toxic shock-like syndrome or necrotizing fasciitis is induced after S. pyogenes infection are not well developed. We demonstrate here that infection with a nonlethal dose of influenza A virus 2 days before intranasal infection with a nonlethal dose of S. pyogenes strains led to a death rate of more than 90% in mice, 10% of which showed necrotizing fasciitis. Infection of lung alveolar epithelial cells by the influenza A virus resulted in viral hemagglutinin expression on the cell surface and promoted internalization of S. pyogenes. However, treatment with monoclonal antibodies to hemagglutinin markedly decreased this internalization. Our results indicate that prior infection with influenza A virus induces a lethal synergism, resulting in the induction of invasive S. pyogenes infection in mice. PMID- 12634368 TI - Characterization of human class-switched polymeric (immunoglobulin M [IgM] and IgA) anti-human immunodeficiency virus type 1 antibodies 2F5 and 2G12. AB - We have previously generated human monoclonal anti-human immunodeficiency virus type 1 (anti-HIV-1) antibodies 2F5IgG and 2G12IgG with an exceptional cross-clade neutralizing potential. 2F5IgG and 2G12IgG passively administrated to macaques were able to confer complete protection from both intravenous and mucosal challenge with pathogenic HIV-simian immunodeficiency virus chimeric strains and have shown beneficial effects in a phase-1 clinical trial. We now class-switched 2F5 and 2G12 to the immunoglobulin M (IgM) or IgA isotype, to enforce features like avidity, complement activation, or the potential to neutralize mucosal transmission. For this purpose we expressed functional polymeric 2F5 and 2G12 antibodies in CHO cells and evaluated their anti-HIV-1 activity in vitro. The class switch had a strong impact on the protective potential of 2F5 and 2G12. 2G12IgM inhibited HIV-1 infection of peripheral blood mononuclear cell cultures up to 28-fold-more efficiently than the corresponding IgG and neutralized all of the primary isolates tested. The 2F5 and 2G12 antibodies of all isotypes were able to interact with active human serum to inhibit viral infection. Furthermore, we demonstrated that polymeric 2F5 and 2G12 antibodies but not the corresponding IgGs could interfere with HIV-1 entry across a mucosal epithelial layer in vitro. Although polymeric 2F5 antibodies had only limited potential in the standard neutralization assay, the results from the mucosal assay suggest that 2F5 and 2G12 antibodies may have a high potential to prevent natural HIV-1 transmission in vivo. PMID- 12634371 TI - Quantitative analysis of herpes simplex virus reactivation in vivo demonstrates that reactivation in the nervous system is not inhibited at early times postinoculation. AB - Recent studies utilizing an ex vivo mouse model of herpes simplex virus (HSV) reactivation have led to the hypothesis that, under physiologic conditions inducing viral reactivation, the immune cells within the infected ganglion block the viral replication cycle and maintain the viral genome in a latent state. One prediction from the ex vivo study is that reactivation in ganglia in vivo would be inhibited at early times postinoculation, when the numbers of inflammatory cells in the ganglia are greatest. To distinguish between an effect of the immune infiltrates on (i) infectious virus produced and/or recovered in the ganglion and (ii) the number of neurons undergoing lytic transcriptional activity (reactivating), an assay to quantify the number of neurons expressing lytic viral protein in ganglia in vivo was developed. Infectious virus and HSV protein positive neurons were quantified from days 9 through 240 postinoculation in latently infected trigeminal ganglia before and at 22 h after hyperthermic-stress induced reactivation. Significant increases in the amount of virus and the number of positive neurons were detected poststress in ganglia at all times examined. Unexpectedly, the greatest levels of reactivation occurred at the times examined most proximal to inoculation. Acyclovir was utilized to stop residual acute-phase virus production, and this treatment did not reduce the level of reactivation on day 14. Thus, the virus measured after induction was a product of reactivation. These data indicate that, in contrast to observations in the ex vivo model, immune cells in the ganglia during the resolution of acute infection do not inhibit reactivation of the virus in ganglia in vivo. PMID- 12634370 TI - Mapping and functional analysis of interaction sites within the cytoplasmic domains of the vaccinia virus A33R and A36R envelope proteins. AB - Incorporation of the vaccinia virus A36R protein into the outer membrane of intracellular enveloped virions (IEV) is dependent on expression of the A33R protein. Possible interactions of the 200-amino-acid cytoplasmic domain of the A36R protein with itself or with the cytoplasmic domain of the A33R, A34R, B5R, or F12L IEV membrane protein was investigated by using the yeast two-hybrid system. A strong interaction was detected only between the cytoplasmic domains of the A36R and A33R proteins. Upon further analyses, the interaction site was mapped to residues 91 to 111 of the A36R protein. To investigate the role of the A36R:A33R interaction during viral infection, five recombinant vaccinia viruses containing B5R-GFP as a marker were constructed. Four had the full-length A36R gene replaced with various-length C-terminal truncations of A36R, of which two contained residues 91 to 111 and two were missing this region. The fifth recombinant virus had an A33R gene with most of the 40-amino-acid cytoplasmic tail deleted. Residues 91 to 111 of A36R and the cytoplasmic tail of A33R were required for a strong interaction between the two proteins during viral infection and for maximal amounts of A36R protein on IEV. Mutants lacking these regions of A33R or A36R formed IEV that exhibited only short sporadic intracellular movement, displayed no actin tails, and formed small plaques on cell monolayers equivalent to those of an A36R deletion mutant and smaller than those formed by point mutations that specifically abrogate actin tail formation. The A33R interaction site of the A36R protein is highly conserved among orthopoxviruses and may overlap binding sites for cellular proteins needed for microtubular movement and actin tail formation. PMID- 12634372 TI - A positive autoregulatory loop of LMP1 expression and STAT activation in epithelial cells latently infected with Epstein-Barr virus. AB - STAT3 and STAT5 are constitutively activated and nuclear in nasopharyngeal carcinoma (NPC) cells. In normal signaling, STATs are only transiently activated. To investigate whether Epstein-Barr virus (EBV), and in particular the protein LMP1, contributes to sustained STAT phosphorylation and activation in epithelial cells, we examined STAT activity in two sets of paired cell lines, HeLa, an EBV converted HeLa cell line, HeLa-Bx1, the NPC-derived cell line CNE2-LNSX, and an LMP1-expressing derivative, CNE2-LMP1. EBV infection was associated with a significant increase in the tyrosine-phosphorylated forms of STAT3 and STAT5 in HeLa-Bx1 cells. This effect correlated with LMP1 expression, since phosphorylated STAT3 and STAT5 levels were also increased in CNE2-LMP1 cells relative to the control CNE2-LNSX cells. No change was observed in STAT1 or STAT6 phosphorylation in these cell lines, nor was there a significant change in the levels of total STAT3, STAT5, STAT1, or STAT6 protein. Tyrosine phosphorylation allows the normally cytoplasmic STAT proteins to enter the nucleus and bind to their recognition sequences in responsive promoters. The ability of LMP1 to activate STAT3 was further established by immunofluorescence assays in which coexpression of LMP1 in transfected cells was sufficient to mediate nuclear relocalization of Flag-STAT3 and by an electrophoretic mobility shift assay which showed that LMP1 expression in CNE2-LNSX cells was associated with increased endogenous STAT3 DNA binding activity. In addition, the activity of a downstream target of STAT3, c Myc, was upregulated in HeLa-Bx1 and CNE2-LMP1 cells. A linkage was established between interleukin-6 (IL-6)- and LMP1-mediated STAT3 activation. Treatment with IL-6 increased phosphorylated STAT3 levels in CNE2-LNSX cells, and conversely, treatment of CNE2-LMP1 cells with IL-6 neutralizing antibody ablated STAT3 activation and c-Myc upregulation. The previous observation that STAT3 activated the LMP1 terminal repeat promoter in reporter assays was extended to show upregulated expression of endogenous LMP1 mRNA and protein in HeLa-Bx1 cells transfected with a constitutively activated STAT3. A model is proposed in which EBV infection of an epithelial cell containing activated STATs would permit LMP1 expression. This in turn would establish a positive feedback loop of IL-6-induced STAT activation, LMP1 and Qp-EBNA1 expression, and viral genome persistence. PMID- 12634373 TI - Interaction with a ubiquitin-like protein enhances the ubiquitination and degradation of hepatitis C virus RNA-dependent RNA polymerase. AB - To identify potential cellular regulators of hepatitis C virus (HCV) RNA dependent RNA polymerase (NS5B), we searched for cellular proteins interacting with NS5B protein by yeast two-hybrid screening of a human hepatocyte cDNA library. We identified a ubiquitin-like protein, hPLIC1 (for human homolog 1 of protein linking intergrin-associated protein and cytoskeleton), which is expressed in the liver (M. F. Kleijnen, A. H. Shih, P. Zhou, S. Kumar, R. E. Soccio, N. L. Kedersha, G. Gill, and P. M. Howley, Mol. Cell 6: 409-419, 2000). In vitro binding assays and in vivo coimmunoprecipitation studies confirmed the interaction between hPLIC1 and NS5B, which occurred through the ubiquitin associated domain at the C terminus of the hPLIC1 protein. As hPLICs have been shown to physically associate with two E3 ubiquitin protein ligases as well as proteasomes (Kleijnen et al., Mol. Cell 6: 409-419, 2000), we investigated whether the stability and posttranslational modification of NS5B were affected by hPLIC1. A pulse-chase labeling experiment revealed that overexpression of hPLIC1, but not the mutant lacking the NS5B-binding domain, significantly shortened the half-life of NS5B and enhanced the polyubiquitination of NS5B. Furthermore, in Huh7 cells that express an HCV subgenomic replicon, the amounts of both NS5B and the replicon RNA were reduced by overexpression of hPLIC1. Thus, hPLIC1 may be a regulator of HCV RNA replication through interaction with NS5B. PMID- 12634374 TI - Hepatitis C virus RNA replication occurs on a detergent-resistant membrane that cofractionates with caveolin-2. AB - The mechanism and machinery of hepatitis C virus (HCV) RNA replication are still poorly understood. In this study, we labeled de novo-synthesized viral RNA in situ with bromouridine triphosphate (BrUTP) in Huh7 cells expressing an HCV subgenomic replicon. By immunofluorescence staining using an anti-BrUTP antibody and confocal microscopy, we showed that the newly synthesized HCV RNA was localized to distinct speckle-like structures, which also contain all of the HCV nonstructural (NS) proteins. These speckles are distinct from lipid droplets and are separated from the endoplasmic reticulum (ER), where some HCV NS proteins also reside. Membrane flotation analysis demonstrated that almost all of the NS5A and part of the NS5B proteins and all of the viral RNA were present in membrane fractions which are resistant to treatment with 1% NP-40 at 4 degrees C. They were cofractionated with caveolin-2, a lipid-raft-associated intracellular membrane protein, in the presence or absence of the detergent. In contrast, the ER-resident proteins were detergent soluble. These properties suggest that the membranes on which HCV RNA replication occurs are lipid rafts recruited from the intracellular membranes. The protein synthesis inhibitors cycloheximide and puromycin did not inhibit viral RNA synthesis, indicating that HCV RNA replication does not require continuous protein synthesis. We suggest that HCV RNA synthesis occurs on a lipid raft membrane structure. PMID- 12634375 TI - nef gene is required for robust productive infection by simian immunodeficiency virus of T-cell-rich paracortex in lymph nodes. AB - The pathogenesis of AIDS virus infection in a nonhuman primate AIDS model was studied by comparing plasma viral loads, CD4(+) T-cell subpopulations in peripheral blood mononuclear cells, and simian immunodeficiency virus (SIV) infection in lymph nodes for rhesus macaques infected with a pathogenic molecularly cloned SIVmac239 strain and those infected with its nef deletion mutant (Deltanef). In agreement with many reports, whereas SIVmac239 infection induced AIDS and depletion of memory CD4(+) T cells in 2 to 3 years postinfection (p.i.), Deltanef infection did not induce any manifestation associated with AIDS up to 6.5 years p.i. To explore the difference in SIV infection in lymphoid tissues, we biopsied lymph nodes at 2, 8, 72, and 82 weeks p.i. and analyzed them by pathological techniques. Maximal numbers of SIV-infected cells (SIV Gag(+), Env(+), and RNA(+)) were detected at 2 weeks p.i. in both the SIVmac239-infected animals and the Deltanef-infected animals. In the SIVmac239-infected animals, most of the infected cells were localized in the T-cell-rich paracortex, whereas in the Deltanef-infected animals, most were localized in B-cell-rich follicles and in the border region between the paracortex and the follicles. Analyses by double staining of CD68(+) macrophages and SIV Gag(+) cells and by double staining of CD3(+) T cells and SIV Env(+) cells revealed that SIV-infected cells were identified as CD4(+) T cells in either the SIVmac239 or the Deltanef infection. Whereas the many functions of Nef protein were reported from in vitro studies, our finding of SIVmac239 replication in the T-cell-rich paracortex in the lymph nodes supports the reported roles of Nef protein in T-cell activation and enhancement of viral infectivity. Furthermore, the abundance of SIVmac239 infection and the paucity of Deltanef infection in the T-cell-rich paracortex accounted for the differences in viral replication and pathogenicity between SIVmac239 and the Deltanef mutant. Thus, our in vivo study indicated that the nef gene enhances SIV replication by robust productive infection in memory CD4(+) T cells in the T-cell-rich region in lymphoid tissues. PMID- 12634376 TI - Mutations in the putative HR-C region of the measles virus F2 glycoprotein modulate syncytium formation. AB - The fusion (F) glycoproteins of measles virus strains Edmonston (MV-Edm) and wtF (MV-wtF) confer distinct cytopathic effects and strengths of hemagglutinin (H) interaction on a recombinant MV-Edm virus. They differ in just two amino acids, V94 and V101 in F-Edm versus M94 and F101 in F-wtF, both of which lie in the relatively uncharacterized F(2) domain. By comparing the sequence of MV F with those of the parainfluenza virus SV5 and Newcastle disease virus (NDV) F proteins, the structures of which are known, we show that MV F(2) also possesses a potential heptad repeat (HR) C domain. In NDV, the N-terminal half of HR-C interacts with HR-A in F(1) while the C-terminal half is induced to kink outward by a central proline residue. We found that this proline is part of an LXP motif conserved in all three viruses. Folding and transport of MV F require this motif to be intact and also require covalent interaction of cysteine residues that probably support the potential HR-A-HR-C interaction. Amino acids 94 and 101, both located in "d" positions of the HR-C helical wheel, lie in the potentially outwardly kinked region. We demonstrate that their effect on MV fusogenicity and glycoprotein interaction is mediated solely by amino acid 94. Substitutions at position 94 with polar or charged amino acids are tolerated poorly or not at all, while changes to smaller and more hydrophilic amino acids are tolerated in both transiently expressed F protein and recombinant virus. MV F V94A and MV F V94G viruses induce extensive syncytium formation and are relatively, or almost completely, resistant to a known inhibitor of MV glycoprotein-induced fusion. We propose that the conformational changes in MV F protein required to expose the fusion peptide involve the C-terminal half of the HR-C helix, specifically amino acid 94. PMID- 12634378 TI - Global changes in Kaposi's sarcoma-associated virus gene expression patterns following expression of a tetracycline-inducible Rta transactivator. AB - An important step in the herpesvirus life cycle is the switch from latency to lytic reactivation. In order to study the life cycle of Kaposi's sarcoma associated herpesvirus (KSHV), we developed a gene expression system in KSHV infected primary effusion lymphoma cells. This system uses Flp-mediated efficient recombination and tetracycline-inducible expression. The Rta transcriptional activator, which acts as a molecular switch for lytic reactivation of KSHV, was efficiently integrated downstream of the Flp recombination target site, and its expression was tightly controlled by tetracycline. Like stimulation with tetradecanoyl phorbol acetate (TPA), the ectopic expression of Rta efficiently induced a complete cycle of viral replication, including a well-ordered program of KSHV gene expression and production of infectious viral progeny. A striking feature of Rta-mediated lytic gene expression was that Rta induced KSHV gene expression in a more powerful and efficient manner than TPA stimulation, indicating that Rta plays a central, leading role in KSHV lytic gene expression. Thus, our streamlined gene expression system provides a novel means not only to study the effects of viral gene products on overall KSHV gene expression and replication, but also to understand the natural viral reactivation process. PMID- 12634379 TI - The ORF45 protein of Kaposi's sarcoma-associated herpesvirus is associated with purified virions. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) ORF45 is encoded by an immediate early gene in the KSHV genome. This protein was recently shown to interact with interferon regulatory factor 7 and inhibit virus-mediated alpha/beta interferon induction (Zhu et al., Proc. Natl. Acad. Sci. USA 99:5573-5578, 2002). ORF45 was characterized as a phosphorylated protein, and it is localized in the cytoplasm of infected cells. In this report, we provide evidence that ORF45 is associated with KSHV virions. (i) ORF45 was detected in gradient-purified virions by Western blotting along with known structural proteins of KSHV including gB, K8.1, and major capsid protein. In contrast, ORF50/Rta, K8alpha, and ORF59/PF8 were not detected in the same virion preparation. (ii) ORF45 comigrates with KSHV virions in sucrose gradient ultracentrifugation. (iii) Virion-associated ORF45 was resistant to trypsin digestion but became sensitive after the virions were treated with detergent which destroys the viral envelope. (iv) ORF45 remained associated with tegument-nucleocapsid complex when virion-specific glycoproteins were removed after detergent treatment. (v) An ORF45 protein band was visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of extensively purified KSHV virions and identified by mass spectrometry. (vi) By immunoelectron microscopy, virus-like structures were specifically stained by anti-ORF45 antibody. Based on the evidence, we conclude that ORF45 is associated with purified KSHV virions and appears to be a tegument protein. The presence of ORF45 in KSHV virions raised the possibility that this protein may be delivered to host cells at the start of infection and therefore have the opportunity to act at the very early stage of the infection, suggesting an important role of ORF45 in KSHV primary infection. PMID- 12634377 TI - A functional YNKI motif in the short cytoplasmic tail of varicella-zoster virus glycoprotein gH mediates clathrin-dependent and antibody-independent endocytosis. AB - The trafficking of varicella-zoster virus (VZV) gH was investigated under both infection and transfection conditions. In initial endocytosis assays performed in infected cells, the three glycoproteins gE, gI, and gB served as positive controls for internalization from the plasma membrane. Subsequently, we discovered that gH in VZV-infected cells was also internalized and followed a similar trafficking pattern. This observation was unexpected because all herpesvirus gH homologues have short endodomains not known to contain trafficking motifs. Further investigation demonstrated that VZV gH, when expressed alone with its chaperone gL, was capable of endocytosis in a clathrin-dependent manner, independent of gE, gI, or gB. Upon inspection of the short gH cytoplasmic tail, we discovered a putative tyrosine-based endocytosis motif (YNKI). When the tyrosine was replaced with an alanine, endocytosis of gH was blocked. Utilizing an endocytosis assay dependent on biotin labeling, we further documented that endocytosis of VZV gH was antibody independent. In control experiments, we showed that gE, gI, and gB also internalized in an antibody-independent manner. Alignment analysis of the VZV gH cytoplasmic tail to other herpesvirus gH homologues revealed two important findings: (i) herpes simplex virus type 1 and 2 homologues lacked an endocytosis motif, while all other alphaherpesvirus gH homologues contained a potential motif, and (ii) the VZV gH and simian varicella virus gH cytoplasmic tails were likely longer in length (18 amino acids) than predicted in the original sequence analyses (12 and 16 amino acids, respectively). The longer tails provided the proper context for a functional endocytosis motif. PMID- 12634380 TI - Limited breadth of a T-helper cell response to a human immunodeficiency virus envelope protein. AB - Single-envelope human immunodeficiency virus (HIV) vaccines have been studied for more than a decade, with some successes in homologous challenge experiments in nonhuman primates but with no clear successes in clinical trials. To gain insight into the breadth of the immunity elicited by such vaccines, we have dissected the T-helper cell response of C57BL/6 mice to an individual, molecularly cloned envelope protein. Here, we report that T-helper cells responsive to HIV type 1 1035 envelope are very highly restricted in C57BL/6 animals: seven different hybridomas recovered from five separate mice recognized the same peptide, PKVSFEPIPIHYCAP, located in the C2 region of gp120. Three of these hybridomas were tested on a natural variant of the peptide but failed to respond. A more extensive analysis of whole splenic populations from other C57BL/6 mice immunized with the 1035 envelope reproducibly confirmed that the gp120-specific T-helper response was almost exclusively focused on a single epitope. We conclude that single-envelope vaccines may frequently fail to provoke an immune response sufficiently diverse to recognize variant sequences among circulating HIV. The results encourage the inclusion of more than one envelope in future vaccines to enhance the potential diversity and respective surveillance capacities of responding T-helper cell populations. PMID- 12634381 TI - Recruitment of polymerase to herpes simplex virus type 1 replication foci in cells expressing mutant primase (UL52) proteins. AB - The ordered assembly of the herpes simplex virus (HSV) type 1 replication apparatus leading to replication compartments likely involves the initial assembly of five viral replication proteins, ICP8, UL9, and the heterotrimeric helicase-primase complex (UL5-UL8-UL52), into replication foci. The polymerase and polymerase accessory protein are subsequently recruited to these foci. Four stages of viral infection (stages I to IV) have been described previously (J. Burkham, D. M. Coen, and S. K. Weller, J. Virol. 72:10100-10107, 1998). Of these, stage III foci are equivalent to the previously described promyelocytic leukemia protein (PML)-associated prereplicative sites and contain all seven replication proteins. We constructed a series of mutations in the putative primase subunit, UL52, of the helicase-primase and have analyzed the mutant proteins for their abilities to form intermediates leading to the formation of replication compartments. The results shown in this paper are consistent with the model that the five proteins, ICP8, UL5, UL8, UL9, and UL52, form a scaffold and that formation of this scaffold does not rely on enzymatic functions of the helicase and primase. Furthermore, we demonstrate that recruitment of polymerase to this scaffold requires the presence of an active primase subunit. These results suggest that polymerase recruitment to replication foci requires primer synthesis. Furthermore, they support the existence of two types of stage III intermediates in the formation of replication compartments: stage IIIa foci, which form the scaffold, and stage IIIb foci, which contain, in addition, HSV polymerase, the polymerase accessory subunit, and cellular factors such as PML. PMID- 12634382 TI - Inoculation of plasmids encoding Japanese encephalitis virus PrM-E proteins with colloidal gold elicits a protective immune response in BALB/c mice. AB - We established a simple and effective method for DNA immunization against Japanese encephalitis virus (JEV) infection with plasmids encoding the viral PrM and E proteins and colloidal gold. Inoculation of plasmids mixed with colloidal gold induced the production of specific anti-JEV antibodies and a protective response against JEV challenge in BALB/c mice. When we compared the efficacy of different inoculation routes, the intravenous and intradermal inoculation routes were found to elicit stronger and more sustained neutralizing immune responses than intramuscular or intraperitoneal injection. After being inoculated twice, mice were found to resist challenge with 100,000 times the 50% lethal dose (LD(50)) of JEV (Beijing-1 strain) even when immunized with a relatively small dose of 0.5 micro g of plasmid DNA. Protective passive immunity was also observed in SCID mice following transfer of splenocytes or serum from plasmid DNA- and colloidal gold-immunized BALB/c mice. The SCID mice resisted challenge with 100 times the LD(50) of JEV. Analysis of histological sections detected expression of proteins encoded by plasmid DNA in the tissues of intravenously, intradermally, and intramuscularly inoculated mice 3 days after inoculation. DNA immunization with colloidal gold elicited encoded protein expression in splenocytes and might enhance immune responses in intravenously inoculated mice. This approach could be exploited to develop a novel DNA vaccine. PMID- 12634383 TI - Epstein-Barr virus nuclear antigen 3C recruits histone deacetylase activity and associates with the corepressors mSin3A and NCoR in human B-cell lines. AB - Epstein-Barr virus (EBV) nuclear antigen 3C (EBNA3C) is a known regulatory transcription factor that has been shown to interact with histone deacetylase 1 (HDAC1) when cotransfected in human cell lines and by in vitro binding experiments. Previous studies have shown that EBNA3C interacts with p300 and prothymosin alpha (ProTalpha) in EBV-infected cells and may be involved in recruiting acetyltransferases to the chromatin for acetylation of histones and transcriptional activation. EBNA3C has also been shown to function as a repressor of transcription when directed to promoters. In this report, we show that EBNA3C complexed with ProTalpha can also recruit deacetylase activity and associates in a complex that includes HDAC1 and HDAC2 in human B cells. A complex of EBNA3C and ProTalpha coimmunoprecipitated with HDAC1 and HDAC2 in cell lines stably expressing EBNA3C. Additionally, this complex associated with the mSin3A and NCoR corepressors in EBNA3C-expressing cell lines and may function in a complex with additional transcription factors known to be repressors of transcription. EBNA3C in complex with ProTalpha recruited deacetylase activity in cell lines stably expressing EBNA3C, and this activity was shown to be partially sensitive to trichostatin A (TSA). This suggests an association with other deacetylases that are insensitive to the general inhibitory effects of TSA, as the entire activity was not abolished in multiple assays. The association between EBNA3C and the corepressors as well as HDACs is likely to depend on the presence of ProTalpha in the complex. Immunoprecipitation with anti-ProTalpha antibody immunoprecipitated EBNA3C and the other repressors, whereas immunoprecipitation with anti-EBNA3C antibody resulted in little or no association with these molecules associated with transcription repression. Clearly, EBNA3C functions as a component of a number of dynamic complexes which function in repression and activation of transcription. PMID- 12634384 TI - The abundant nuclear enzyme PARP participates in the life cycle of simian virus 40 and is stimulated by minor capsid protein VP3. AB - The abundant nuclear enzyme poly(ADP-ribose) polymerase (PARP) functions in DNA damage surveillance and repair and at the decision between apoptosis and necrosis. Here we show that PARP binds to simian virus 40 (SV40) capsid proteins VP1 and VP3. Furthermore, its enzymatic activity is stimulated by VP3 but not by VP1. Experiments with purified mutant proteins demonstrated that the PARP binding domain in VP3 is localized to the 35 carboxy-terminal amino acids, while a larger peptide of 49 amino acids was required for full stimulation of its activity. The addition of 3-aminobenzamide (3-AB), a known competitive inhibitor of PARP, demonstrated that PARP participates in the SV40 life cycle. The titer of SV40 propagated on CV-1 cells was reduced by 3-AB in a dose-dependent manner. Additional experiments showed that 3-AB did not affect viral DNA replication or capsid protein production. PARP did not modify the viral capsid proteins in in vitro poly(ADP-ribosylation) assays, implying that it does not affect SV40 infectivity. On the other hand, it greatly reduced the magnitude of the host cytopathic effects, a hallmark of SV40 infection. Additional experiments suggested that the stimulation of PARP activity by VP3 leads the infected cell to a necrotic pathway, characterized by the loss of membrane integrity, thus facilitating the release of mature SV40 virions from the cells. Our studies identified a novel function of the minor capsid protein VP3 in the recruitment of PARP for the SV40 lytic process. PMID- 12634385 TI - Selective virus resistance conferred by expression of Borna disease virus nucleocapsid components. AB - Persistent viral infections can render host cells resistant to superinfection with closely related viruses by largely uncharacterized mechanisms. We present evidence for superinfection exclusion in brains of Borna disease virus (BDV) infected rats and in persistently infected Vero cells, and we suggest that acquired resistance to BDV is due to unbalanced intracellular levels of viral nucleocapsid components. We observed that expression of BDV protein P, N, or X rendered human cells resistant to subsequent challenge with BDV but not with other RNA viruses, indicating that incorrect stoichiometry of nucleocapsid components selectively blocked the polymerase activity of incoming viruses. Vero cells containing high levels of an untranslatable BDV-N transcript remained virus susceptible, demonstrating that viral protein rather than RNA mediated resistance. Transient overexpression of BDV-P in persistently infected Vero cells was also remarkably effective against BDV, indicating that the intracellular balance of viral nucleocapsid components could serve as a target for future therapeutic antiviral strategies. PMID- 12634386 TI - Three-dimensional localization of pORF65 in Kaposi's sarcoma-associated herpesvirus capsid. AB - Of the six herpesvirus capsid proteins, the smallest capsid proteins (SCPs) share the least sequence homology among herpesvirus family members and have been implicated in virus specificity during infection. The herpes simplex virus-1 (HSV 1) SCP was shown to be horn shaped and to specifically bind the upper domain of each major capsid protein in hexons but not in pentons. In Kaposi's sarcoma associated herpesvirus (KSHV), the protein encoded by the ORF65 gene (pORF65) is the putative SCP but its location remains controversial due to the absence of such horn-shaped densities from both the pentons and hexons of the KSHV capsid reconstructions. To directly locate the KSHV SCP, we have used electron cryomicroscopy and three-dimensional reconstruction techniques to compare the three-dimensional structure of KSHV capsids to that of anti-pORF65 antibody labeled capsids. Our difference map shows prominent antibody densities bound to the tips of the hexons but not to pentons, indicating that KSHV SCP is attached to the upper domain of the major capsid protein in hexons but not to that in pentons, similar to HSV-1 SCP. The lack of horn-shaped densities on the hexons indicates that KSHV SCP exhibits structural features that are substantially different from those of HSV-1 SCP. The location of SCP at the outermost regions of the capsid suggests a possible role in mediating capsid interactions with the tegument and cytoskeletal proteins during infection. PMID- 12634388 TI - Kaposi's sarcoma-associated herpesvirus cytotoxic T lymphocytes recognize and target Darwinian positively selected autologous K1 epitopes. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is the infectious cause of Kaposi's sarcoma (KS) and certain lymphoproliferations particularly in the context of human immunodeficiency virus (HIV) type 1-induced immunosuppression. The introduction of effective therapies to treat HIV has led to a decline in the incidence of KS, suggesting that immune responses may play a role in controlling KSHV infection and pathogenesis. Cytotoxic-T-lymphocyte (CTL) activity against KSHV proteins has been demonstrated; however, the identification of KSHV CTL epitopes remains elusive and problematic. Although the herpesvirus genomic layout is generally conserved, KSHV encodes a unique hypervariable protein, K1, with intense biological selection pressure at specific amino acid sites. To investigate whether this variability is partly driven by cellular immunity, we designed K1 peptides that match only the unique viral sequence for every individual studied here (autologous peptides). We identified functional CTL epitopes within K1's most variable areas, and we show that a given individual responds only to autologous peptides and not to peptides from other individuals. Furthermore, these epitopes are highly conserved sequences within KSHV isolates from a specific strain but are not conserved between different strains. We conclude that CTL recognition contributes to K1, and therefore to KSHV, evolution. PMID- 12634387 TI - The B subunit of Escherichia coli heat-labile enterotoxin enhances CD8+ cytotoxic T-lymphocyte killing of Epstein-Barr virus-infected cell lines. AB - Epstein-Barr virus (EBV) is associated with a number of important human cancers, including nasopharyngeal carcinoma, gastric carcinoma, and Hodgkin's lymphoma. These tumors express a viral nuclear antigen, EBV nuclear antigen 1 (EBNA1), which cannot be presented to T cells in a major histocompatibility complex class I context, and the viral latent membrane proteins (LMPs). Although the LMPs are expressed in these tumors, no effective immune response is made. We report here that exposure to the cholera-like enterotoxin B subunit (EtxB) in EBV-infected lymphoblastoid cell lines (LCLs) enhances their susceptibility to killing by LMP specific CD8(+) cytotoxic T lymphocytes (CTLs) in a HLA class I-restricted manner. CTL killing of LCLs is dramatically increased through both transporter associated protein-dependent and -independent epitopes after EtxB treatment. The use of mutant B subunits revealed that the enhanced susceptibility of LCLs to CTL killing is dependent on the B subunit's interaction with GM(1) but not its signaling properties. These important findings could underpin the development of novel approaches to treating EBV-associated malignancies and may offer a general approach to increasing the presentation of other tumor and viral antigens. PMID- 12634390 TI - In silico pattern-based analysis of the human cytomegalovirus genome. AB - More than 200 open reading frames (ORFs) from the human cytomegalovirus genome have been reported as potentially coding for proteins. We have used two pattern based in silico approaches to analyze this set of putative viral genes. With the help of an objective annotation method that is based on the Bio-Dictionary, a comprehensive collection of amino acid patterns that describes the currently known natural sequence space of proteins, we have reannotated all of the previously reported putative genes of the human cytomegalovirus. Also, with the help of MUSCA, a pattern-based multiple sequence alignment algorithm, we have reexamined the original human cytomegalovirus gene family definitions. Our analysis of the genome shows that many of the coded proteins comprise amino acid combinations that are unique to either the human cytomegalovirus or the larger group of herpesviruses. We have confirmed that a surprisingly large portion of the analyzed ORFs encode membrane proteins, and we have discovered a significant number of previously uncharacterized proteins that are predicted to be G-protein coupled receptor homologues. The analysis also indicates that many of the encoded proteins undergo posttranslational modifications such as hydroxylation, phosphorylation, and glycosylation. ORFs encoding proteins with similar functional behavior appear in neighboring regions of the human cytomegalovirus genome. All of the results of the present study can be found and interactively explored online (http://cbcsrv.watson.ibm.com/virus/). PMID- 12634389 TI - CK2 protein kinase is stimulated and redistributed by functional herpes simplex virus ICP27 protein. AB - It has been shown previously (S. Wadd, H. Bryant, O. Filhol, J. E. Scott, T.-T. Hsieh, R. D. Everett, and J. B. Clements, J. Biol. Chem. 274:28991-28998, 2000) that ICP27, an essential and multifunctional herpes simplex virus type 1 (HSV-1) protein, interacts with CK2 and with heterogeneous ribonucleoprotein K (hnRNP K). CK2 is a pleiotropic and ubiquitous protein kinase, and the tetrameric holoenzyme consists of two catalytic alpha or alpha' subunits and two regulatory beta subunits. We show here that HSV-1 infection stimulates CK2 activity. CK2 stimulation occurs at early times after infection and correlates with redistribution of the holoenzyme from the nucleus to the cytoplasm. Both CK2 stimulation and redistribution require expression and cytoplasmic accumulation of ICP27. In HSV-1-infected cells, CK2 phosphorylates ICP27 and affects its cytoplasmic accumulation while it also phosphorylates hnRNP K, which is not ordinarily phosphorylated by this kinase, suggesting an alteration of hnRNP K activities. This is the first example of CK2 stimulation by a viral protein in vivo, and we propose that it might facilitate the HSV-1 lytic cycle by, for example, regulating trafficking of ICP27 protein and/or viral RNAs. PMID- 12634391 TI - Characterization of the complete genome of the Tupaia (tree shrew) adenovirus. AB - The members of the family Adenoviridae are widely spread among vertebrate host species and normally cause acute but innocuous infections. Special attention is focused on adenoviruses because of their ability to transform host cells, their possible application in vector technology, and their phylogeny. The primary structure of the genome of Tupaia adenovirus (TAV), which infects Tupaia spp. (tree shrew) was determined. Tree shrews are taxonomically assumed to be at the base of the phylogenetic tree of mammals and are frequently used as laboratory animals in neurological and behavior research. The TAV genome is 33,501 bp in length with a G+C content of 49.96% and has 166-bp inverted terminal repeats. Analysis of the complete nucleotide sequence resulted in the identification of 109 open reading frames (ORFs) with a coding capacity of at least 40 amino acid residues. Thirty-eight of them are predicted to encode viral proteins based on the presence of transcription and translation signals and sequence and positional conservation. Thirty viral ORFs were found to show significant similarities to known adenoviral genes, arranged into discrete early and late genome regions as they are known from mastadenoviruses. Analysis of the nucleotide content of the TAV genome revealed a significant CG dinucleotide depletion at the genome ends that suggests methylation of these genomic regions during the viral life cycle. Phylogenetic analysis of the viral gene products, including penton and hexon proteins, viral protease, terminal protein, protein VIII, DNA polymerase, protein IVa2, and 100,000-molecular-weight protein, revealed that the evolutionary lineage of TAV forms a separate branch within the phylogenetic tree of the Mastadenovirus genus. PMID- 12634392 TI - Engineering the transmissible gastroenteritis virus genome as an expression vector inducing lactogenic immunity. AB - The genome of the coronavirus transmissible gastroenteritis virus (TGEV) has been engineered as an expression vector with an infectious cDNA. The vector led to the efficient (>40 micro g/10(6) cells) and stable (>20 passages) expression of a heterologous gene (green fluorescent protein [GFP]), driven by the transcription regulating sequences (TRS) of open reading frame (ORF) 3a inserted in the site previously occupied by the nonessential ORFs 3a and 3b. Expression levels driven by this TRS were higher than those of an expression cassette under the control of regulating sequences engineered with the N gene TRS. The recombinant TGEV including the GFP gene was still enteropathogenic, albeit with a 10- to 10(2) fold reduction in enteric tissue growth. Interestingly, a specific lactogenic immune response against the heterologous protein has been elicited in sows and their progeny. The engineering of an additional insertion site for the heterologous gene between viral genes N and 7 led to instability and to a new genetic organization of the 3' end of the recombinant viruses. As a consequence, a major species of subgenomic mRNA was generated from a TRS with the noncanonical core sequence 5'-CUAAAA-3'. Extension of the complementarity between the TRS and sequences at the 3' end of the viral leader was associated with transcriptional activation of noncanonical core sequences. The engineered vector led to expression levels as high as those of well-established vectors and seems very promising for the development of vaccines and, possibly, for gene therapy. PMID- 12634393 TI - Intracellular assembly and secretion of recombinant subviral particles from tick borne encephalitis virus. AB - It is believed that flavivirus assembly occurs by intracellular budding of the nucleocapsid into the lumen of the endoplasmic reticulum (ER). Recombinant expression of tick-borne encephalitis (TBE) virus envelope proteins prM and E in mammalian cells leads to their incorporation into enveloped recombinant subviral particles (RSPs), which have been used as a model system for studying assembly and entry processes and are also promising vaccine candidates. In this study, we analyzed the formation and secretion of TBE virus RSPs and of a membrane anchor free E homodimer in mammalian cells. Immunofluorescence microscopy showed that E was accumulated in the lumen of the ER. RSPs were observed by electron microscopy in the rough and smooth ER and in downstream compartments of the secretory pathway. About 75% of the particles appeared to be of the size expected for RSPs (about 30 nm in diameter), but a number of larger particles and tubular structures were also observed in these compartments. Secretion of membrane anchor free E dimers was detected 30 min after synthesis of prM and E, and secretion of RSPs was detected 1 h after synthesis of prM and E. We also found that the presence of the single N-linked oligosaccharide side chain on the E protein and its trimming by glucosidases was necessary for secretion of RSPs and truncated E dimers. Our results suggest that incorporation of prM and E into RSPs occurs at the ER membrane without other viral elements being required, followed by rapid transport along the compartments of the secretory pathway and secretion. Moreover, the carbohydrate side chain of E is involved in at least one assembly or transport step. PMID- 12634394 TI - Apoptotic cells, including macrophages, are prominent in Theiler's virus-induced inflammatory, demyelinating lesions. AB - Theiler's murine encephalomyelitis virus (TMEV) persists in the mouse central nervous system principally in macrophages, and infected macrophages in culture undergo apoptosis. We have detected abundant apoptotic cells in perivascular cuffs and inflammatory, demyelinating lesions of SJL mice chronically infected with TMEV. T cells comprised 74% of apoptotic cells, while 8% were macrophages, 0.6% were astrocytes, and approximately 17% remained unidentified. In situ hybridization revealed viral RNA in approximately 1% of apoptotic cells. PMID- 12634395 TI - Interleukin-7-dependent production of RANTES that correlates with human immunodeficiency virus disease progression. AB - There is a relationship between CD4-T-cell number and circulating interleukin 7 (IL-7) levels in human immunodeficiency virus (HIV)-positive individuals. Here, we show that IL-7 induced a dose-dependent production of CCL3 (MIP-1alpha), CCL4 (MIP-1beta), and CCL5 (RANTES) in peripheral blood mononuclear cells (PBMC), ex vivo tonsil lymphoid tissue of HIV(-) individuals, and PBMC from HIV(+) individuals, suggesting that IL-7 may regulate beta-chemokine production in vivo. In a cross-sectional study of HIV(+) individuals (n = 130), a weak but significant correlation between IL-7 and RANTES was noted (r = 0.379; P < 0.001). Remarkably, the correlation between IL-7 and RANTES increased to an r value of 0.798 (P < 0.001) if individuals with low CD4 cell counts (<200 cells/ micro l) were excluded from the analysis. Our results suggest that there is a relationship between IL-7 and the production of RANTES both in vitro and in vivo that is lost in immune-compromised patients (CD4 count of <200 cells/ micro l) but that could be restored by antiretroviral therapy. Unlike the case for IL-7, high levels of RANTES suggest an intermediate stage of HIV disease progression. PMID- 12634396 TI - Role of CD8(+) lymphocytes in control and clearance of measles virus infection of rhesus monkeys. AB - The creation of an improved vaccine for global measles control will require an understanding of the immune mechanisms of measles virus containment. To assess the role of CD8(+) cytotoxic T lymphocytes in measles virus clearance, rhesus monkeys were depleted of CD8(+) lymphocytes by monoclonal anti-CD8 antibody infusion and challenged with wild-type measles virus. The CD8(+) lymphocyte depleted animals exhibited a more extensive rash, higher viral loads at the peak of virus replication, and a longer duration of viremia than did the control antibody-treated animals. These findings indicate a central role for CD8(+) lymphocytes in the control of measles virus infections and the importance of eliciting a cell-mediated immune response in new measles vaccine strategies. PMID- 12634397 TI - Cytoskeletal requirements for hepatitis C virus (HCV) RNA synthesis in the HCV replicon cell culture system. AB - Hepatitis C virus (HCV) induces microtubule aggregates in infected hepatocytes. To determine if cytoskeletal elements are important for HCV RNA synthesis, we examined the effect of cytoskeleton inhibitors on HCV replicon transcription in Huh7 cells. The data demonstrate that HCV replication complex-mediated RNA synthesis requires microtubule and actin polymerization. PMID- 12634398 TI - Disassembly of human immunodeficiency virus type 1 cores in vitro reveals association of Nef with the subviral ribonucleoprotein complex. AB - The human immunodeficiency virus type 1 (HIV-1) virulence factor Nef enhances viral infectivity in single-cycle infection assays and accelerates HIV-1 replication in vitro. It has been reported that the effects of Nef are mediated early after viral entry and before the completion of reverse transcription, as viral DNA synthesis is strongly attenuated during infection by Nef-defective virions. Our previous work has demonstrated that Nef is associated with mature HIV-1 cores, implicating Nef in the regulation of HIV-1 core stability. Here we report a comparative analysis of HIV-1 cores isolated from wild-type and Nef defective particles. We observed no effect of Nef on HIV-1 core structure or stability; however, Nef cosedimented with a subviral ribonucleoprotein complex following dissociation of CA. These results indicate that Nef interacts tightly with an internal component of the HIV-1 core. They further suggest that virion associated Nef may facilitate an early step in HIV-1 infection following dissociation of the viral capsid in the target cell. PMID- 12634399 TI - Unexpected absence of the Epstein-Barr virus (EBV) lyLMP-1 open reading frame in tumor virus isolates: lack of correlation between Met129 status and EBV strain identity. AB - The lytic cycle-associated lytic latent membrane protein-1 (lyLMP-1) of Epstein Barr virus (EBV) is an amino-terminally truncated form of the oncogenic LMP-1. Although lyLMP-1 shares none of LMP-1's transforming and signal transducing activities, we recently reported that lyLMP-1 can negatively regulate LMP-1 stimulated NF-kappaB activation. The lyLMP-1 protein encoded by the B95-8 strain of EBV initiates from methionine 129 (Met129) of the LMP-1 open reading frame (ORF). The recent report that Met129 in the B95-8 LMP-1 ORF is not conserved in the Akata strain of EBV prompted us to screen a panel of EBV-positive cell lines for conservation of Met129 and lyLMP-1 expression. We found that 15 out of 16 tumor-associated virus isolates sequenced encoded an ATT or ACC codon in place of ATG in the LMP-1 ORF at position 129, and tumor cell lines harboring isolates lacking an ATG at codon 129 did not express the lyLMP-1 protein. In contrast, we found that EBV DNA from 22 out of 37 healthy seropositive donors retained the Met129 codon. Finally, the lyLMP-1 initiator occurs variably within distinct EBV strains and its presence cannot be predicted by EBV strain identity. Thus, Met129 is not peculiar to the B95-8 strain of EBV, but rather can be found in the background of several evolutionarily distinct EBV strains. Its absence from EBV isolates from tumors raises the possibility of selective pressure on Met129 in EBV-dependent tumors. PMID- 12634400 TI - Latency of viral expression in vivo is not related to CpG methylation in the U3 region and part of the R region of the long terminal repeat of bovine leukemia virus. AB - Bovine leukemia virus (BLV) is silent in most cells detectable in vivo, and the repression of its expression allows BLV to evade the host's immune response. In this study, we examined whether CpG methylation of DNA might be involved in the regulation of the expression of BLV in vivo. To investigate the effects of CpG methylation on the activity of the long terminal repeat (LTR) of BLV, we measured the transactivation activity of this region after treatment with the CpG methyltransferase SssI by using a luciferase reporter system. The activity of methylated LTR was significantly lower than that of nonmethylated LTR. Therefore, we examined the extent of CpG methylation of the U3 region and part of the R region of the LTR in BLV-infected cattle and in experimentally BLV-infected sheep at various clinical stages by the bisulfite genomic sequencing method. We detected no or minimal CpG methylation at all stages examined in cattle and sheep, and our results indicate that CpG methylation probably does not participate in the silencing of BLV in vivo. PMID- 12634401 TI - Treatment of human immunodeficiency virus type 1 virions depleted of cyclophilin A by natural endogenous reverse transcription restores infectivity. AB - We have previously shown that virions with nef deleted can be restored to wild type infectivity by treatment to induce natural endogenous reverse transcription (NERT). Since Nef and cyclophilin A (CyPA) appear to act in similar ways on postentry events, we determined whether NERT treatment would restore infectivity to virions depleted of CyPA. Our results show that the infectivity of virions depleted of CyPA by treatment with cyclosporine A could be restored by NERT treatment, while mutants in the CyPA binding loop of capsid could only be partially restored. These results suggest that CyPA is involved in some aspect of the uncoating process. PMID- 12634402 TI - Human coronavirus 229E: receptor binding domain and neutralization by soluble receptor at 37 degrees C. AB - Truncated human coronavirus HCoV-229E spike glycoproteins containing amino acids 407 to 547 bound to purified, soluble virus receptor, human aminopeptidase N (hAPN). Soluble hAPN neutralized the infectivity of HCoV-229E virions at 37 degrees C, but not 4 degrees C. Binding of hAPN may therefore trigger conformational changes in the viral spike protein at 37 degrees C that facilitate virus entry. PMID- 12634403 TI - Epstein-Barr virus regulates STAT1 through latent membrane protein 1. AB - This study shows a mechanism for the increase of signal transducer and activator of transcription 1 (STAT1) in Epstein-Barr virus-immortalized cells. Latent membrane protein 1 (LMP-1) expression was sufficient to induce STAT1 expression, DNA binding, and transcriptional activity. LMP-1-expressing cells can induce an increase in STAT1 expression in LMP-negative cells in the same culture, suggesting an indirect regulation of STAT1 expression. The increase in STAT1 expression is mediated by the C-terminal activating region 1 (CTAR-1) and/or CTAR 2 domains of LMP-1 and is inhibited by mutant IkappaB, demonstrating a role for NFkappaB in LMP-1-mediated STAT1 expression. PMID- 12634404 TI - Polyprotein processing protease of African swine fever virus: purification and biochemical characterization. AB - The purified recombinant African swine fever virus polyprotein processing protease cleaves the two GG-X sites in polyprotein pp62 with the same efficiency. Cleavage at the site that is first recognized in vivo is not a requisite for cleavage at the second site, suggesting the existence of mechanisms that control the ordered processing of the polyprotein during infection. PMID- 12634406 TI - Use of transgenic HLA A*0201/Kb and HHD II mice to evaluate frequency of cytomegalovirus IE1-derived peptide usage in eliciting human CD8 cytokine response. AB - Unlike the pp65 protein of human cytomegalovirus (CMV), which has an immunodominant peptide, pp65(495-503), recognized by human CD8(+) cells in the context of HLA A*0201, the fine peptide specificity for CMV IE1 has shown no such immunodominance. With the use of transgenic HLA A*0201/Kb and HHD II mice, a selected pool of IE1 peptides, including IE1(p256-264), IE1(p297-304), and IE1(p316-324), were shown to stimulate cytolytic T-lymphocyte lysis in the context of HLA A*0201. Based on an intracellular gamma interferon response, IE1(p297-304), a previously unrecognized CD8 epitope, triggered a prominent response to CMV IE1 in HLA A*0201 subjects. PMID- 12634407 TI - Diverse dengue type 2 virus populations contain recombinant and both parental viruses in a single mosquito host. AB - Envelope (E) protein genes sampled from populations of dengue 2 (DEN-2) virus in individual Aedes aegypti mosquitoes and in serum from dengue patients were copied to cDNA, cloned, and sequenced. The nucleotide sequences of the E genes in more than 70% of the clones differed from the consensus sequence for the corresponding virus population at up to 11 sites, and 24 of the 94 clones contained at least one stop codon. Virus populations recovered up to 2 years apart yielded clones with similar polymorphisms in the E gene. For one mosquito, the clones obtained fell into two genotypes. One group of sequences was closely related to those of viruses recovered from dengue patients in the same locality (Yangon, Myanmar) since 1995 and were classified as Asian 1 genotype. The second group were Cosmopolitan genotype viruses which were also circulating in Yangon in 2000 and which were related to DEN-2 viruses sampled from southern China in 1999. Finally, one clone was identified as a recombinant genome composed of portions of these two "parental" genotypes. This is the first report of recombinant and parental dengue viruses in a single host. PMID- 12634408 TI - Transcriptional regulators and myelopoiesis: the role of serum response factor and CREB as targets of cytokine signaling. AB - Hematopoiesis is a complex process in which mature myeloid and lymphoid cells are produced from a small population of pluripotent stem cells within the bone marrow. Blood cell formation occurs, in part, by progenitor cell exposure to humoral growth regulators, known as hematopoietic cytokines, as well as by the regulated expression of genes by transcription factors. In this paper, we review two important nuclear proteins, the serum response factor and the cyclic adenosine monophosphate response element-binding protein, as downstream targets of mitogens, with a specific focus on hematopoietic cytokine signaling and the role these proteins play in gene regulation. PMID- 12634409 TI - Human adult marrow cells support prolonged expansion of human embryonic stem cells in culture. AB - Prolonged propagation of human embryonic stem (hES) cells is currently achieved by coculture with primary mouse embryonic fibroblasts (MEFs) serving as feeder cells. Unlike mouse ES cells, adding growth factors such as leukemia inhibitory factor is insufficient to maintain undifferentiated hES cells without feeder cells. The presence of uncharacterized rodent cells or crude extracts imposes a risk to the clinical applications of hES cells. While others looked for a replacement of MEFs with human fetal cells, we attempted to use easily accessible postnatal human cells such as human marrow stromal cells (hMSCs). Culture expanded hMSCs from multiple donors were used as feeder cells to support growth of the H1 hES cell line under a serum-free culture condition. Human ES cell colonies cultured on irradiated hMSCs amplified >100-fold during the 30-day continuous culture (in five passages). The longest continuous expansion of hES cells on hMSCs tested to date is 13 passages. The expanded hES cells displayed the unique morphology and molecular markers characteristic of undifferentiated hES cells as observed when they were cultured on MEFs. They expressed the transcription factor Oct-4, a membrane alkaline phosphatase, and the stage specific embryonic antigen (SSEA)-4, but not the SSEA-1 marker. Expanded hES cells on hMSCs retained unique differentiation potentials in culture and a normal diploid karyotype. The well-studied hMSCs (and this animal cell- and serum-free system) may provide a clinically and ethically feasible method to expand hES cells for novel cell therapies. In addition, this system may help to identify cytokines and adhesion molecules that are required for the self-renewal of hES cells. PMID- 12634405 TI - The CCR5 and CXCR4 coreceptors are both used by human immunodeficiency virus type 1 primary isolates from subtype C. AB - Human immunodeficiency virus type 1 (HIV-1) subtype C viruses with different coreceptor usage profiles were isolated from 29 South African patients with advanced AIDS. All 24 R5 isolates were inhibited by the CCR5-specific agents, PRO 140 and RANTES, while the two X4 viruses and the three R5X4 viruses were sensitive to the CXCR4-specific inhibitor, AMD3100. The five X4 or R5X4 viruses were all able to replicate in peripheral blood mononuclear cells that did not express CCR5. When tested using coreceptor-transfected cell lines, one R5 virus was also able to use CXCR6, and another R5X4 virus could use CCR3, BOB/GPR15, and CXCR6. The R5X4 and X4 viruses contained more-diverse V3 loop sequences, with a higher overall positive charge, than the R5 viruses. Hence, some HIV-1 subtype C viruses are able to use CCR5, CXCR4, or both CXCR4 and CCR5 for entry, and they are sensitive to specific inhibitors of entry via these coreceptors. These observations are relevant to understanding the rapid spread of HIV-1 subtype C in the developing world and to the design of intervention and treatment strategies. PMID- 12634410 TI - Differences between peripheral blood and cord blood in the kinetics of lineage restricted hematopoietic cells: implications for delayed platelet recovery following cord blood transplantation. AB - Cord blood (CB) cells are a useful source of hematopoietic cells for transplantation. The hematopoietic activities of CB cells are different from those of bone marrow and peripheral blood (PB) cells. Platelet recovery is significantly slower after transplantation with CB cells than with cells from other sources. However, the cellular mechanisms underlying these differences have not been elucidated. We compared the surface marker expression profiles of PB and CB hematopoietic cells. We focused on two surface markers of hematopoietic cell immaturity, i.e., CD34 and AC133. In addition to differences in surface marker expression, the PB and CB cells showed nonidentical differentiation pathways from AC133(+)CD34(+) (immature) hematopoietic cells to terminally differentiated cells. The majority of the AC133(+)CD34(+) PB cells initially lost AC133 expression and eventually became AC133(-)CD34(-) cells. In contrast, the AC133(+)CD34(+) CB cells did not go through the intermediate AC133(-)CD34(+) stage and lost both markers simultaneously. Meanwhile, the vast majority of megakaryocyte progenitors were of the AC133(-)CD34(+) phenotype. We conclude that the delayed recovery of platelets after CB transplantation is due to both subpopulation distribution and the process of differentiation from AC133(+)CD34(+) cells. PMID- 12634411 TI - Multilineage potential of homozygous stem cells derived from metaphase II oocytes. AB - Human stem cells derived from human fertilized oocytes, fetal primordial germ cells, umbilical cord blood, and adult tissues provide potential cell-based therapies for repair of degenerating or damaged tissues. However, the diversity of major histocompatibility complex (MHC) antigens in the general population and the resultant risk of immune-mediated rejection complicates the allogenic use of established stem cells. We assessed an alternative approach, employing chemical activation of nonfertilized metaphase II oocytes for producing stem cells homozygous for MHC. By using F1 hybrid mice (H-2-B/D), we established stem cell lines homozygous for H-2-B and H-2-D, respectively. The undifferentiated cells retained a normal karyotype, expressed stage-specific embryonic antigen-1 and Oct4, and were positive for alkaline phosphatase and telomerase. Teratomatous growth of these cells displayed the development of a variety of tissue types encompassing all three germ layers. In addition, these cells demonstrated the potential for in vitro differentiation into endoderm, neuronal, and hematopoietic lineages. We also evaluated this homozygous stem cell approach in human tissue. Five unfertilized blastocysts were derived from a total of 25 human oocytes, and cells from one of the five hatched blastocysts proliferated and survived beyond two passages. Our studies demonstrate a plausible "homozygous stem cell" approach for deriving pluripotent stem cells that can overcome the immune-mediated rejection response common in allotransplantation, while decreasing the ethical concerns surrounding human embryonic stem cell research. PMID- 12634412 TI - Colocalization analysis of sialomucins CD34 and CD164. AB - Flow cytometric protocols are employed to identify and characterize hemopoietic stem/progenitor populations before transplantation. Cell surface antigens, including CD34, are employed in this process and widely used in harvest protocols, which largely ignores the potential functional role of such antigens. Transmembrane glycoprotein sialomucins, including CD34 and CD164, have been implicated in cell-to-cell interactions and activation. CD164, also expressed on early hemopoietic populations, was reported to have a possible function facilitating CD34(+) cells to adhere to bone marrow stroma. In this study, we employed high-definition laser-scanning confocal microscopy to investigate CD34 and CD164 surface co-localization patterns on bone marrow and cord blood cells and to compare the expression patterns using a three-dimensional computer generated method developed in house. Differential interference microscopy analysis revealed bone marrow membrane activity was higher than the corresponding cord blood counterpart, perhaps indicating the marrow microenvironmental nature. Fluorescence analysis of CD34 and CD164 antigens showed both were expressed first in a halo-like pattern and second in antigen-dense pockets. Three-dimensional computer analyses further revealed that this pocketing corresponded to dense crest-like surface structures appearing to rise from the point of adherence on the slide. Further, it was found that CD34 and CD164 display strong colocalization patterns on cells expressing both antigens. The dual nature of the CD34 and CD164 antigens discovered here lends further evidence to the previous literature implicating a strong functional link between these two sialomucins, which should be considered in the transplantation arena and in the function of such sialomucins as negative regulators of cell proliferation. PMID- 12634413 TI - Potential use of embryonic stem cells for the treatment of mouse parkinsonian models: improved behavior by transplantation of in vitro differentiated dopaminergic neurons from embryonic stem cells. AB - BACKGROUND: and Aims. The purpose of the present study was to examine the efficacy of transplantation of mouse embryonic-stem-(ES)-cell-derived tyrosine hydroxylase-positive (TH(+)) cells into Parkinsonian mice using behavioral tests and immunohistochemical evaluation. METHODS: Undifferentiated ES cells carrying the enhanced green fluorescent protein (EGFP) gene were differentiated into a cell population containing TH(+) neurons using a five-step in vitro differentiation method. These ES-cell-derived cells were used as allografts in Parkinsonian mice, made by administering injections of 6-hydroxydopamine (6 OHDA). Fifteen hemiparkinsonian mice were divided into three groups. Four weeks after 6-OHDA injection, mice in groups 1, 2, and 3 received phosphate-buffered saline, 1 x 10(4) graft cells, and 1 x 10(5) graft cells, respectively, into their dopamine-denervated striata. RESULTS: Improved rotational behavior was observed in the graft-transplanted groups (groups 2 and 3) 2 weeks after transplantation. Mice in group 2 displayed a continuous maintenance of reduced rotational behavior, while those in group 3 showed ipsilateral rotation toward the lesioned side at 4, 6, and 8 weeks after transplantation. Tumor formation was observed in one mouse in group 3. TH(+) cells were found at the grafted sites 8 weeks after transplantation in mice in groups 2 and 3, some of which were immunopositive to GFP, demonstrating the presence of dopaminergic neurons derived from the ES cells. CONCLUSION: Transplantation of in vitro differentiated ES cells changed rotational behavior in Parkinsonian mice. Our results suggest the potential availability of ES cells for Parkinson's disease. PMID- 12634414 TI - Migration and differentiation of myogenic precursors following transplantation into the developing rat brain. AB - There is increasing evidence that muscle-derived precursor cells can, under appropriate conditions, give rise to other than myogenic cell types. Transplantation into the embryonic ventricular zone provides a unique opportunity to study the migration and differentiation of non-neural somatic progenitor cells in response to instructive cues within the developing neuroepithelium. Here, we demonstrate that myogenic cell lines grafted into the ventricles of rat embryos showed widespread migration into several host brain compartments. In contrast to incorporation patterns observed after transplantation of neural cells, grafted myoblasts incorporated virtually exclusively along endogenous blood vessels. Preferential incorporation sites included cortex, olfactory bulb, hippocampus, striatum, thalamus, hypothalamus, and tectum. While the engrafted myoblasts showed no evidence of neural differentiation, a fraction exhibited pronounced coexpression of endothelial marker antigens. These findings support the concept of a close developmental relationship between the myogenic and the endothelial lineages. Used as a delivery system, transfected myoblasts may be exploited for widespread gene transfer to the perivascular compartment of the perinatal central nervous system. PMID- 12634415 TI - Multipotential mesenchymal stem cells from femoral bone marrow near the site of osteonecrosis. AB - Stem cell-based therapies for degenerative disorders and injuries are promising in the new era. Multipotential mesenchymal stem cells (MSCs) from bone marrow (BM) are on the leading edge because they are easy to expand in culture while maintaining their multilineage potential. In vitro assessment of the chondrogenic and osteogenic potentials of cultured MSCs has been established, and the BM used in those experiments was exclusively from healthy donors via iliac crest aspiration. It is unknown whether human marrow obtained from femurs also contains these multipotential MSCs. We collected marrow from proximal femurs of two patients undergoing total hip replacement surgery for femoral head osteonecrosis and isolated and culture expanded MSCs to about 20 population doublings. These cells were homogeneously positive for beta1-integrin. When pelleted into aggregates and cultured in a medium containing transforming growth factor-beta3 for 14 days, the cells began to express mRNA for aggrecan and collagen type II and to deposit immunoreactive collagen type II and sulfated proteoglycans in the matrix, hallmarks of chondrogenic differentiation. These MSCs could also be differentiated into osteocytic lineage in vitro, as shown by increased expression of alkaline phosphatase activity and deposition of mineral content onto culture plates. These results indicate that femoral BM obtained during hip surgeries also contained multipotential MSCs. These data imply that direct replacement therapy using MSCs from in situ marrow may be possible in the future and that an MSC bank may be established by using marrow from this approach, bypassing the necessity for iliac marrow aspiration from healthy donors. PMID- 12634416 TI - Cardiac allograft acceptance after localized bone marrow transplantation by isolated limb perfusion in nonmyeloablated recipients. AB - Donor-specific tolerance to cardiac grafts may be induced by hematopoietic chimerism. This study evaluates the potential of localized bone marrow transplantation (BMT) performed by isolated limb (IL) perfusion to induce tolerance to secondary cardiac grafts without myeloablative conditioning. BALB/c recipients (H2d) preconditioned with lethal and sublethal doses of busulfan were injected i.v. and IL with 10(7) whole bone marrow cells (wBMCs) from B10 donors (H2(b)). Two hours after IL infusion of PKH-labeled wBMCs into myeloablated hosts, there were few labeled cells in the host peripheral blood (p < 0.001 versus i.v.) and femurs of the infused limb contained 57% +/- 7% PKH-labeled blasts (p < 0.001 versus 8% +/- 0.6% after i.v.). Femurs of the noninfused limbs contained 60-70 PKH-labeled blasts (p < 0.001 versus i.v.-BMT) after 2 days and 47% +/- 5% of 0.32 x 10(7) donor cells (p < 0.001 versus 78% +/- 4% of 1.2 x 10(7) donor cells in infused femurs) after 4 weeks. The survival rates of myeloablated hosts were 90% and 80% after i.v. and IL infusion, respectively, and the chimeras had 78%-84% donor peripheral blood cells. In recipients conditioned with 35 mg/g busulfan, the levels of donor chimerism in peripheral blood were 33% +/- 4% and 21% +/- 4% at 3 weeks after i.v.- and IL-BMT, respectively. Transplantation of donor-matched (H2(b)) secondary vascularized hearts in these chimeras after 3 weeks resulted in graft survival for periods exceeding 8 weeks, while third-party (H2(k)) allografts were acutely rejected (p < 0.001 versus H2(b)). These data indicate that IL perfusion is a reliable alternative procedure for establishment of hematopoietic chimerism and donor-specific tolerance without myeloablative conditioning. PMID- 12634417 TI - Engraftment capacity of umbilical cord blood cells processed by either whole blood preparation or filtration. AB - Umbilical cord blood (UCB) preparation needs to be optimized in order to develop more simplified procedures for volume reduction, as well as to reduce the amount of contaminating cells within the final stem cell transplant. We evaluated a novel filter device (StemQuick((TM))E) and compared it with our routine buffy coat (BC) preparation procedure for the enrichment of hematopoietic progenitor cells (HPCs). Two groups of single or pooled UCB units were filtered (each n = 6), or equally divided in two halves and processed by filtration and BC preparation in parallel (n = 10). The engraftment capacity of UCB samples processed by whole blood (WB) preparation was compared with paired samples processed by filtration in the nonobese diabetic/severe combined immunodeficient (NOD/SCID) mouse animal model. Filtration of UCB units in the two groups with a mean volume of 87.8 and 120.7 ml, respectively, and nucleated cell (NC) content of 9.7 and 23.8 x 10(8) resulted in a sufficient mean cell recovery for mononucleated cells ([MNCs] 74.2%-77.5%), CD34(+) cells (76.3%-79.0%), and colony forming cells (64.1%-86.3%). Moreover, we detected a relevant depletion of the transplants for RBCs (89.2%-90.0%) and platelets ([PLTs] 77.5%-86.1%). In contrast, the mean depletion rate using BC processing proved to be significantly different for PLTs (10%, p = 0.03) and RBCs (39.6%, p < 0.01). The NC composition showed a highly significant increase in MNCs and a decrease in granulocytes after filtration (p < 0.01), compared with a less significant MNC increase in the BC group (p < 0.05). For mice transplanted with WB-derived progenitors, we observed a mean of 15.3% +/- 15.5% of human CD45(+) cells within the BM compared with 19.9% +/- 16.8% for mice transplanted with filter samples (p = 0.03). The mean percentage of human CD34(+) cells was 4.2% +/- 3.1% for WB samples and 4.5% +/- 3.2% for filter samples (p = 0.68). As the data of NOD/SCID mice transplantation demonstrated a significant engraftment capacity of HPCs processed by filtration, no negative effect on the engraftment potential of filtered UCB cells versus non volume-reduced cells from WB transplants was found. The StemQuick((TM))E filter devices proved to be a useful tool for Good Manufacturing Practices conform enrichment of HPCs and MNCs out of UCB. Filtration enables a quick and standardized preparation of a volume-reduced UCB transplant, including a partial depletion of granulocytes, RBCs, and PLTs without the need for centrifugation. Therefore, it seems very probable that filter-processed UCB transplants will also result in sufficient hematopoietic reconstitution in humans. PMID- 12634418 TI - Human umbilical cord blood as a source of transplantable hepatic progenitor cells. AB - Human umbilical cord blood (UCB) cells have many advantages as grafts for cell transplantation because of the immaturity of newborn cells compared with adult cells. In contrast to their hematopoietic and mesenchymal potential, it remains unclear whether UCB cells have endodermal competence. Here, with a view to utilize UCB cells for cell transplantation into injured liver, we investigated the hepatic potential of UCB cells both in vitro and in vivo. We determined the most efficient conditions leading UCB cells to produce albumin (ALB). In a novel primary culture system supplemented with a combination of growth/differentiation factors, about 50% of UCB cells in 21-day cultures expressed ALB, and the ALB(+) cells coexpressed hepatocyte lineage markers. The ALB-expressing cells were able to proliferate in the culture system. Moreover, in the cell-transplantation model into liver-injured severe combined immunodeficient mice, inoculated UCB cells developed into functional hepatocytes in the liver, which released human ALB into the sera of the recipient mice. In conclusion, this study demonstrates that human UCB is a source of transplantable hepatic progenitor cells. Our findings may have relevance to clinical application of UCB-derived cell transplantation as a novel therapeutic option for liver failure. PMID- 12634419 TI - Adherent cells generated during long-term culture of human umbilical cord blood CD34+ cells have characteristics of endothelial cells and beneficial effect on cord blood ex vivo expansion. AB - Hematopoiesis depends on the association of hematopoietic stem cells with stromal cells that constitute the hematopoietic microenvironment. The in vitro development of the endothelial cell from umbilical cord blood (UCB) is not well established and has met very limited success. In this study, UCB CD34(+) cells were cultured for 5 weeks in a stroma-free liquid culture system using thrombopoietin, flt3 ligand, and granulocyte-colony stimulating factor. By week 4 5, we found that firmly adherent fibroblast-like cells were established. These cells showed characteristics of endothelial cells expressing von Willebrand factor, human vascular cell adhesion molecule-1, human intracellular adhesion molecule-1, human CD31, E-selectin, and human macrophage. Furthermore, when comparing an ex vivo system without an established endothelial monolayer to an ex vivo system with an established endothelial monolayer, better expansion of total nucleated cells, CD34(+) cells, and colony-forming units (CFUs)-granulocyte macrophage and CFUs-granulocyte-erythroid-megakaryocyte-macrophage were found during culture. This phenomenon was in part due to the fact that a significant reduction of apoptotic fractions was found in the CD34(+) cells, which were cultured on the adherent monolayer for up to 5 weeks. To gather quantitative data on the number of endothelial cells derived from a given number of CD34 cells, we performed limiting dilution assay by using Poisson distribution: the number of tested cells (linear scale) producing a 37% negative culture (logarithmic scale) is the number of cells containing one endothelial cell. By this method, one endothelial cell may be found from 314 CD34(+) cells after 5 weeks of culture. These results suggest that the UCB CD34(+) cell fraction contains endothelial cell precursors, establishing the hematopoietic microenvironment and providing the beneficial effects through downregulating apoptosis on UCB expansion protocols. These observations may provide insight for future cellular therapy or graft engineering. PMID- 12634420 TI - Interleukin-3 promotes proliferation and differentiation of human hematopoietic stem cells but reduces their repopulation potential in NOD/SCID mice. AB - In the present study we explored systematically the influence of human interleukin-3 (IL-3) on the cord blood (CB) cell-derived production of human hematopoietic cells in the bone marrow, blood, and spleen of chimeric nonobese/severe combined immunodeficient mice ((NOD/SCID) mice. CB mononuclear cells and MACS-enriched CB CD34(+) cells were injected into irradiated NOD/SCID mice. The mice were additionally transplanted with a stably transfected rat fibroblast cell line expressing the human IL-3 gene (Rat-IL-3) constitutively, or with the nontransfected rat fibroblast cell line as a control (Rat-1). Rat-IL-3 mice displayed a higher engraftment of human hematopoietic cells in bone marrow, spleen, and peripheral blood compared with mice with Rat-1 cotransplantation. When we transplanted their total bone marrow cell population into secondary mice, surprisingly, mice transplanted with bone marrow cells from Rat-1 mice displayed a higher proportion of human hematopoietic cells compared with Rat-IL-3 mice. As expected, bone marrow cultures (BMCs) from Rat-IL-3 mice contained a higher proportion of human cells than Rat-1 bone marrow cells. However, when BMCs were passaged to new flasks, we observed a higher proportion of human cells in BMCs from Rat-1 mice compared with BMCs from Rat-IL-3 mice. IL-3 promotes the proliferation and differentiation of hematopoietic stem cells in chimeric bone marrow. In addition, IL-3 may play a role in the depletion of hematopoietic stem cells in chimeric bone marrow. In the absence of IL-3, the hematopoietic stem cells may remain in a quiescent state and proliferation can be induced by stimuli, including secondary transplantation or cell passage. PMID- 12634422 TI - Variable character of O-O and M-O bonding in side-on (eta(2)) 1:1 metal complexes of O2. AB - The structures and the O-O and M-O bonding characters of a series of reported side-on (eta(2)) 1:1 metal complexes of O(2) are analyzed by using density functional theory calculations. Comparison of the calculated and experimental systems with respect to O-O bond distance, O-O stretching frequency, and O-O and M-O bond orders provides new insights into subtle influences relevant to O(2) activation processes in biology and catalysis. The degree of charge transfer from the generally electron-rich metals to the dioxygen fragment is found to be variable, such that there are species well described as superoxides, others well described as peroxides, and several cases having intermediate character. Increased charge transfer to dioxygen takes place via overlap of the metal d(xy) orbital with the in-plane pi* orbital of O(2) and results in increased M-O bond orders and decreased O-O bond orders. Comparison of theory and experiment over the full range of compounds studied suggests that reevaluation of the O-O bond lengths determined from certain x-ray crystal structures is warranted; in one instance, an x-ray crystal structure redetermination was performed at low temperature, confirming the theoretical prediction. Librational motion of the coordinated O(2) is identified as a basis for significant underestimation of the O-O distance at high temperature. PMID- 12634421 TI - Insulin-degrading enzyme regulates the levels of insulin, amyloid beta-protein, and the beta-amyloid precursor protein intracellular domain in vivo. AB - Two substrates of insulin-degrading enzyme (IDE), amyloid beta-protein (Abeta) and insulin, are critically important in the pathogenesis of Alzheimer's disease (AD) and type 2 diabetes mellitus (DM2), respectively. We previously identified IDE as a principal regulator of Abeta levels in neuronal and microglial cells. A small chromosomal region containing a mutant IDE allele has been associated with hyperinsulinemia and glucose intolerance in a rat model of DM2. Human genetic studies have implicated the IDE region of chromosome 10 in both AD and DM2. To establish whether IDE hypofunction decreases Abeta and insulin degradation in vivo and chronically increases their levels, we characterized mice with homozygous deletions of the IDE gene (IDE --). IDE deficiency resulted in a >50% decrease in Abeta degradation in both brain membrane fractions and primary neuronal cultures and a similar deficit in insulin degradation in liver. The IDE - mice showed increased cerebral accumulation of endogenous Abeta, a hallmark of AD, and had hyperinsulinemia and glucose intolerance, hallmarks of DM2. Moreover, the mice had elevated levels of the intracellular signaling domain of the beta amyloid precursor protein, which was recently found to be degraded by IDE in vitro. Together with emerging genetic evidence, our in vivo findings suggest that IDE hypofunction may underlie or contribute to some forms of AD and DM2 and provide a mechanism for the recently recognized association among hyperinsulinemia, diabetes, and AD. PMID- 12634423 TI - Purified particulate methane monooxygenase from Methylococcus capsulatus (Bath) is a dimer with both mononuclear copper and a copper-containing cluster. AB - Particulate methane monooxygenase (pMMO) is a membrane-bound enzyme that catalyzes the oxidation of methane to methanol in methanotropic bacteria. Understanding how this enzyme hydroxylates methane at ambient temperature and pressure is of fundamental chemical and potential commercial importance. Difficulties in solubilizing and purifying active pMMO have led to conflicting reports regarding its biochemical and biophysical properties, however. We have purified pMMO from Methylococcus capsulatus (Bath) and detected activity. The purified enzyme has a molecular mass of approximately 200 kDa, probably corresponding to an alpha(2)beta(2)gamma(2) polypeptide arrangement. Each 200-kDa pMMO complex contains 4.8 +/- 0.8 copper ions and 1.5 +/- 0.7 iron ions. Electron paramagnetic resonance spectroscopic parameters corresponding to 40-60% of the total copper are consistent with the presence of a mononuclear type 2 copper site. X-ray absorption near edge spectra indicate that purified pMMO is a mixture of Cu(I) and Cu(II) oxidation states. Finally, extended x-ray absorption fine structure data are best fit with oxygennitrogen ligands and a 2.57-A Cu-Cu interaction, providing direct evidence for a copper-containing cluster in pMMO. PMID- 12634424 TI - A gain-of-function mutation in ftsA bypasses the requirement for the essential cell division gene zipA in Escherichia coli. AB - ZipA and FtsA are recruited independently to the FtsZ cytokinetic ring (Z ring) and are essential for cell division of Escherichia coli. The molecular role of FtsA in cell division is unknown; however, ZipA is thought to stabilize the Z ring, anchor it to the membrane, and recruit downstream cell division proteins. Here we demonstrate that the requirement for ZipA can be bypassed completely by a single alteration in a conserved residue of FtsA (FtsA*). Cells with ftsA* in single copy in place of WT ftsA or with ftsA* alone on a multicopy plasmid divide mostly normally, whether they are zipA+ or zipA-. Experiments with ftsQAZ and ftsQA*Z on multicopy plasmids indicate that ftsQAZzipA+ and ftsQA*ZzipA- cells divide fairly normally, whereas ftsQAZzipA- cells divide poorly and ftsQA*ZzipA+ cells display a phenotype that suggests their septa are unusually stable. In support of the idea that ftsA* stabilizes Z rings, single-copy ftsA* confers resistance to excess MinC, which destabilizes Z rings. The inhibitory effect of excess ZipA on division is also suppressed by ftsA*. These results suggest that the molecular mechanism of the FtsA* bypass is to stabilize FtsZ assembly via a parallel pathway and that FtsA* can replace the multiple functions of ZipA. This is an example of a complete functional replacement of an essential prokaryotic cell division protein by another and may explain why most bacteria can divide without an obvious ZipA homolog. PMID- 12634425 TI - Opening protein pores with chaotropes enhances Fe reduction and chelation of Fe from the ferritin biomineral. AB - Iron is concentrated in ferritin, a spherical protein with a capacious cavity for ferric nanominerals of <4,500 Fe atoms. Global ferritin structure is very stable, resisting 6 M urea and heat (85 degrees C) at neutral pH. Eight pores, each formed by six helices from 3 of the 24 polypeptide subunits, restrict mineral access to reductant, protons, or chelators. Protein-directed transport of Fe and aqueous Fe(3+) chemistry (solubility approximately 10(-18) M) drive mineralization. Ferritin pores are "gated" based on protein crystals and Fe chelation rates of wild-type (WT) and engineered proteins. Pore structure and gate residues, which are highly conserved, thus should be sensitive to environmental changes such as low concentrations of chaotropes. We now demonstrate that urea or guanidine (1-10 mM), far below concentrations for global unfolding, induced multiphasic rate increases in Fe(2+)-bipyridyl formation similar to conservative substitutions of pore residues. Urea (1 M) or the nonconservative LeuPro substitution that fully unfolded pores without urea both induced monophasic rate increases in Fe(2+) chelation rates, indicating unrestricted access between mineral and reductantchelator. The observation of low melting ferritin subdomains by CD spectroscopy (melting midpoint 53 degrees C), accounting for 10% of ferritin alpha-helices, is unprecedented. The low-melting ferritin subdomains are pores, based on percentage helix and destabilization by either very dilute urea solutions (1 mM) or LeuPro substitution, which both increased Fe(2+) chelation. Biological molecules may have evolved to control gating of ferritin pores in response to cell iron need and, if mimicked by designer drugs, could impact chelation therapies in iron-overload diseases. PMID- 12634426 TI - A solution to the worn tooth conundrum in primate functional anatomy. AB - Worn teeth are a bane to paleobiologists interested in the diets of human ancestors and other fossil primates. Although worn teeth dominate fossil assemblages, their shapes are usually not used to reconstruct the diets of extinct species. The problem is that traditional studies of primate dental functional anatomy have focused on unworn morphology. This has limited most functional analyses to only a few well-represented fossil species. This paper introduces a method to characterize and compare worn occlusal morphology in primates using laser scanning and geographic information systems technologies. A study of variably worn chimpanzee and gorilla molars indicates that differences between these species in tooth shape remain consistent at given stages of wear. Although cusp slope decreases with wear in both taxa, angularity values remain unchanged. These results indicate that African ape teeth wear in a manner that keeps them mechanically efficient for fracturing specific foods. Studies of changes in tooth shape with wear add a new dimension to dental functional anatomy, and offer a more complete picture of dental-dietary adaptations. Also, given how rare unworn teeth are in the fossil record, the ability to include worn specimens in analyses opens the door to reconstructing the diets of many more extinct primate groups, allowing us to better understand the adaptive radiation of our order. PMID- 12634427 TI - Demonstration of functional coupling between gamma -aminobutyric acid (GABA) synthesis and vesicular GABA transport into synaptic vesicles. AB - l-Glutamic acid decarboxylase (GAD) exists as both membrane-associated and soluble forms in the mammalian brain. Here, we propose that there is a functional and structural coupling between the synthesis of gamma-aminobutyric acid (GABA) by membrane-associated GAD and its packaging into synaptic vesicles (SVs) by vesicular GABA transporter (VGAT). This notion is supported by the following observations. First, newly synthesized [(3)H]GABA from [(3)H]l-glutamate by membrane-associated GAD is taken up preferentially over preexisting GABA by using immunoaffinity-purified GABAergic SVs. Second, the activity of SV-associated GAD and VGAT seems to be coupled because inhibition of GAD also decreases VGAT activity. Third, VGAT and SV-associated Ca(2+)calmodulin-dependent kinase II have been found to form a protein complex with GAD. A model is also proposed to link the neuronal stimulation to enhanced synthesis and packaging of GABA into SVs. PMID- 12634429 TI - Floating plant dominance as a stable state. AB - Invasion by mats of free-floating plants is among the most important threats to the functioning and biodiversity of freshwater ecosystems ranging from temperate ponds and ditches to tropical lakes. Dark, anoxic conditions under thick floating plant cover leave little opportunity for animal or plant life, and they can have large negative impacts on fisheries and navigation in tropical lakes. Here, we demonstrate that floating-plant dominance can be a self-stabilizing ecosystem state, which may explain its notorious persistence in many situations. Our results, based on experiments, field data, and models, represent evidence for alternative domains of attraction in ecosystems. An implication of our findings is that nutrient enrichment reduces the resilience of freshwater systems against a shift to floating-plant dominance. On the other hand, our results also suggest that a single drastic harvest of floating plants can induce a permanent shift to an alternative state dominated by rooted, submerged growth forms. PMID- 12634428 TI - Promyogenic members of the Ig and cadherin families associate to positively regulate differentiation. AB - Determination and differentiation of cells in the skeletal muscle lineage is positively regulated by cell-cell contact. Cell-surface proteins proposed to mediate this effect include both classical cadherins and Ig superfamily members; potential interactions between the promyogenic activities of these classes of protein, however, are unknown. We show here that CDO and BOC, two promyogenic Ig superfamily members that bind to each other in a cis fashion, form complexes with N- and M-cadherin. These complexes contain beta-catenin and are enriched at sites of cell-cell contact between myoblasts. In transient expression assays, the ectodomains and intracellular regions of CDO, BOC, and N-cadherin each interact independently, suggesting that the interactions occur in a cis fashion; consistent with this conclusion, cadherin-mediated cell adhesion is not required for them to occur. Stable expression in myoblasts of a CDO deletion mutant deficient in its ability to associate with N-cadherin interferes with differentiation as assessed by biochemical, morphological, and reporter gene assays, suggesting that this interaction is functionally important in myogenesis. Thus, some of the cell-cell contact-mediated activities that are required for myogenesis seem to be based on interdependent activities of promyogenic classical cadherins and Ig superfamily members. PMID- 12634430 TI - The human obesity gene map: the 2002 update. AB - This is the ninth update of the human obesity gene map, incorporating published results through October 2002 and continuing the previous format. Evidence from single-gene mutation obesity cases, Mendelian disorders exhibiting obesity as a clinical feature, quantitative trait loci (QTLs) from human genome-wide scans and various animal crossbreeding experiments, and association and linkage studies with candidate genes and other markers is reviewed. For the first time, transgenic and knockout murine models exhibiting obesity as a phenotype are incorporated (N = 38). As of October 2002, 33 Mendelian syndromes relevant to human obesity have been mapped to a genomic region, and the causal genes or strong candidates have been identified for 23 of these syndromes. QTLs reported from animal models currently number 168; there are 68 human QTLs for obesity phenotypes from genome-wide scans. Additionally, significant linkage peaks with candidate genes have been identified in targeted studies. Seven genomic regions harbor QTLs replicated among two to five studies. Attempts to relate DNA sequence variation in specific genes to obesity phenotypes continue to grow, with 222 studies reporting positive associations with 71 candidate genes. Fifteen such candidate genes are supported by at least five positive studies. The obesity gene map shows putative loci on all chromosomes except Y. More than 300 genes, markers, and chromosomal regions have been associated or linked with human obesity phenotypes. The electronic version of the map with links to useful sites can be found at http://obesitygene.pbrc.edu. PMID- 12634431 TI - Relationship of serum adiponectin and leptin concentrations with body fat distribution in humans. AB - OBJECTIVE: We investigated whether serum concentrations of adiponectin are determined by body fat distribution and compared the findings with leptin. RESEARCH METHODS AND PROCEDURES: Serum concentrations of adiponectin and leptin were measured by radioimmunoassay (n = 394) and analyzed for correlation with sex, age, and body fat distribution, i.e., waist-to-hip ratio, waist and hip circumference, and subcutaneous adipose tissue area of the lower leg as assessed by magnetic resonance imaging. RESULTS: After adjusting for sex and percentage of body fat, adiponectin was negatively (r = -0.17, p < 0.001) and leptin was positively (r = 0.22, p < 0.001) correlated with waist-to-hip ratio. Leptin, but not adiponectin, correlated with both waist (r = 0.49, p < 0.001) and hip circumference (r = 0.46, p < 0.001). Furthermore, leptin, but not adiponectin, correlated with the proportion of subcutaneous fat of the lower leg cross sectional area (r = 0.37, p < 0.001). DISCUSSION: These data suggest that both adipocytokines are associated with central body fat distribution, and serum adiponectin concentrations are determined predominantly by the visceral fat compartment. PMID- 12634432 TI - Does relative leptinemia predict weight gain in humans? PMID- 12634433 TI - Calcium modulation of adiposity. PMID- 12634434 TI - Genetic variation in the leptin receptor gene, leptin, and weight gain in young Dutch adults. AB - OBJECTIVE: To investigate the association between leptin levels, polymorphisms in the leptin receptor (LEPR) gene, and weight gain. RESEARCH METHODS AND PROCEDURES: From two large prospective cohorts in The Netherlands (n = 17,500), we compared the baseline leptin of 259 subjects who had gained an average of 12.6 kg (range 5.5 to 33 kg) with 277 subjects who kept stable weight (range -2.6 to 3.1 kg) after a mean follow-up of 6.8 years. Three polymorphisms in the LEPR gene (Lys109Arg, Gln223Arg, and Lys656Asn) were determined. RESULTS: Weight gainers had significantly higher baseline leptin levels than those who kept stable weight (odds ratio = 1.27, 95% confidence interval 1.1 to 1.5, per SD increase in log(e) transformed leptin). Weight gainers with the Arg109 or the Arg223 alleles had higher leptin levels compared with the noncarriers of these alleles. Only among men, the association between leptin and weight gain tended to be stronger among those with an Arg223 allele compared with those without this mutation. DISCUSSION: Relatively high leptin levels predict weight gain, suggesting that leptin resistance plays a role in the development of obesity in the general population. Higher leptin levels for those with a Lys109Arg or Gln223Arg mutation (or a linked other marker) may imply that these subjects have a modified functional leptin receptor. However, the role of these mutations on weight gain is limited. PMID- 12634435 TI - High dietary calcium reduces body fat content, digestibility of fat, and serum vitamin D in rats. AB - OBJECTIVE: This study investigated which aspect of energy balance was responsible for the decrease in body fat content of rats fed a high-calcium, high-dairy protein diet. RESEARCH METHODS AND PROCEDURES: Male Wistar rats were fed a control diet (25% kcal fat, 14% kcal protein from casein, 0.4% by weight calcium) or high-calcium diet (25% kcal fat, 7% kcal protein from nonfat dry milk, 7% kcal protein from casein, 2.4% calcium) for 85 days. Body weights, digestible energy intakes, energy expenditures, rectal temperatures, body composition, and serum glucose, insulin, free fatty acids, triglycerides, and 1,25-dihydroxyvitamin D were measured. RESULTS: Rats fed high-calcium diet gained significantly less weight than controls and had 29% less carcass fat. Gross energy intake was not significantly different between groups, but digestible energy was 90% of gross energy in the high-calcium diet compared with 94% in the control diet because of increased fecal excretion of dietary lipid. The difference in digestible energy intake accounted for differences in carcass energy. Body temperatures and energy expenditures of the rats were not different. The high-calcium diet reduced serum triglycerides by 23% and serum 1,25-dihydroxyvitamin D by 86%. DISCUSSION: These results confirm that a high-calcium diet decreases body weight and fat content due to a lower digestible energy intake caused by increased fecal lipid and a nonsignificant reduction in gross energy intake. PMID- 12634437 TI - Enhanced erythrocyte adhesiveness/aggregation in obesity corresponds to low-grade inflammation. AB - OBJECTIVE: Previous studies have suggested that obesity enhances the inflammatory response, producing macromolecules involved in the induction and/or maintenance of increased erythrocyte aggregation. The objectives of this study were to evaluate the correlation between inflammation markers, erythrocyte adhesiveness/aggregation, and the degree of obesity and to assess phosphatidylserine expression on erythrocyte surface membrane of obese vs. nonobese individuals. RESEARCH METHODS AND PROCEDURES: Erythrocyte adhesiveness/aggregation in the peripheral venous blood was evaluated by using a new biomarker, phosphatidylserine expression was assessed by means of flow cytometry, and markers of inflammation were measured in 65 subjects: 30 obese [body mass index (BMI) = 41 +/- 7.7 kg/m(2)] and 35 nonobese (BMI = 24 +/- 2.7 kg/m(2)) individuals. Pearson correlations and Student's t test were performed. RESULTS: A highly significant difference was noted in the degree of erythrocyte adhesiveness/aggregation and markers of inflammation between the study groups. BMI correlated with erythrocyte adhesiveness/aggregation (r = 0.42, p = 0.001), erythrocyte sedimentation rate (r = 0.42, p = 0.001), high-sensitive C-reactive protein (r = 0.55, p < 10(-4)), fibrinogen (r = 0.37, p = 0.004), and white blood cell count (r = 0.45, p < 10(-4)). The degree of erythrocyte adhesiveness/aggregation correlated with erythrocyte sedimentation rate (r = 0.5, p < 10(-4)), high-sensitive C-reactive protein (r = 0.56, p < 10(-4)), fibrinogen (r = 0.54, p < 10(-4)), and white blood cell count (r = 0.32, p = 0.01). DISCUSSION: Our results suggest that obesity-related erythrocyte adhesiveness/aggregation is probably mediated through increased concentrations of adhesive macromolecules in the circulation and not necessarily through hyperlipidemia or phosphatidylserine exposure on erythrocyte's membrane. PMID- 12634436 TI - Medium-chain triglycerides increase energy expenditure and decrease adiposity in overweight men. AB - OBJECTIVE: The objectives of this study were to compare the effects of diets rich in medium-chain triglycerides (MCTs) or long-chain triglycerides (LCTs) on body composition, energy expenditure, substrate oxidation, subjective appetite, and ad libitum energy intake in overweight men. RESEARCH METHODS AND PROCEDURES: Twenty four healthy, overweight men with body mass indexes between 25 and 31 kg/m(2) consumed diets rich in MCT or LCT for 28 days each in a crossover randomized controlled trial. At baseline and after 4 weeks of each dietary intervention, energy expenditure was measured using indirect calorimetry, and body composition was analyzed using magnetic resonance imaging. RESULTS: Upper body adipose tissue (AT) decreased to a greater extent (p < 0.05) with functional oil (FctO) compared with olive oil (OL) consumption (-0.67 +/- 0.26 kg and -0.02 +/- 0.19 kg, respectively). There was a trend toward greater loss of whole-body subcutaneous AT volume (p = 0.087) with FctO compared with OL consumption. Average energy expenditure was 0.04 +/- 0.02 kcal/min greater (p < 0.05) on day 2 and 0.03 +/- 0.02 kcal/min (not significant) on day 28 with FctO compared with OL consumption. Similarly, average fat oxidation was greater (p = 0.052) with FctO compared with OL intake on day 2 but not day 28. DISCUSSION: Consumption of a diet rich in MCTs results in greater loss of AT compared with LCTs, perhaps due to increased energy expenditure and fat oxidation observed with MCT intake. Thus, MCTs may be considered as agents that aid in the prevention of obesity or potentially stimulate weight loss. PMID- 12634438 TI - Regulation of resistin by gonadal, thyroid hormone, and nutritional status. AB - OBJECTIVE: Resistin was recently identified as a hormone secreted by adipocytes that is under hormonal and nutritional control. This hormone has been suggested to be the link between obesity and type 2 diabetes. The aim of this study was to assess the influence of gender, gonadal status, thyroid hormones, pregnancy, and food restriction on resistin mRNA levels in adipose tissue of rats. RESEARCH METHODS AND PROCEDURES: We have determined resistin mRNA expression by Northern blot analysis in all experimental sets. RESULTS: Resistin mRNA expression is influenced by age, with the highest hormone levels existing at 45 days after birth and decreasing thereafter. Resistin mRNA expression is higher in men than in women. Moreover, we studied the effect of orchidectomy and ovariectomy in rats of different ages and showed that gonadal hormones increase adipose tissue resistin mRNA expression in male rats. Resistin is also regulated by thyroid hormones; it is severely decreased in hyperthyroid rats. Our results clearly show that chronic food restriction (30% of ad libitum food intake) led to a decrease in adipose tissue mRNA levels in normal cycling female rats and pregnant rats. In pregnancy, resistin mRNA levels were enhanced particularly at midgestation. DISCUSSION: Our observations indicate that resistin is influenced by gender, gonadal status, thyroid hormones, and pregnancy. These findings suggest that resistin could explain the decreased insulin sensitivity during puberty and could be the link between sex steroids and insulin sensitivity. Moreover, resistin could mediate the effect of thyroid hormones on insulin resistance and the state of insulin resistance present during pregnancy. PMID- 12634439 TI - Phenylethanolamine N-methyltransferase G-148A genetic variant and weight loss in obese women. AB - OBJECTIVE: To understand the impact of the phenylethanolamine N-methyltransferase (PNMT) G-148A gene and nutritional variables on weight loss in obese women. RESEARCH METHODS AND PROCEDURES: One hundred forty-nine women, ages 45 to 65 with a body mass index of >30 kg/m(2), participated in a 6-month, open-label intervention that included sibutramine (15 mg/d) and a monthly health-education class. Anthropometric measurements, vital signs, food frequency, exercise log, medication compliance, and psychological and sociological questionnaires were completed each month. Genetic polymorphisms of PNMT were determined. RESULTS: Univariate analysis of G/G, G/A, and A/A genotypes against tertiles of percentage of weight loss were significant at 3 but not at 6 months (Pearson chi(2): p < 0.006; homozygous/heterozygosity: p < 0.002, p < 0.253, and p < 0.122, respectively). A regression model that included the PNMT genetic variation and certain nutrition and exercise variables demonstrated that only the PNMT gene (beta = 0.360, SE 0.585, and p = 0.003) was statistically significant at 6 months, and the total calories (beta = -0.925, SE = 0.004, and p = 0.009), fiber intake (beta = 0.621, SE = 0.124, and p = 0.000), and PNMT (beta = 0.262, SE = 1.415, and p = 0.024) were significant. DISCUSSION: The homozygosity/heterozygosity of the PNMT gene was highly predictive of significant weight loss with sibutramine during the first 3 months, which highlights the need for specific pharmacotherapy. The early weight-loss success of those subjects who were homozygous for PNMT may have motivated and selected those that would make further dietary changes, which then augmented their final weight loss. PMID- 12634440 TI - Objectively measured physical activity in a representative sample of 3- to 4-year old children. AB - OBJECTIVE: This study aimed to describe levels of physical activity in a representative sample of preschool children and to quantify tracking of activity over 1 year. RESEARCH METHODS AND PROCEDURES: Physical activity (mean accelerometry counts/minute) was assessed over 3 days using the Computer Science and Applications accelerometer in 3- to 4-year-old children (n = 104; 52 boys; mean age, 3.7 +/- 0.4 years). In 60 children (30 boys), measurements were repeated 1 year later. RESULTS: Mean total activity at baseline was 777 +/- 207 counts/minute in boys and 657 +/- 172 counts/minute for girls; this gender difference was significant (p < 0.001). In the cross-sectional analysis, total activity was significantly positively related to age (r = 0.37, p = 0.007). In the sample followed up for 1 year, mean total activity was 849 +/- 252. The longitudinal analysis confirmed that total physical activity increased over the 1 year period (paired Student's t test, p < 0.001). The tracking rank order correlation coefficient of total activity count over 1 year was r = 0.40 (p < 0.001). DISCUSSION: This study suggests that total activity increases during the preschool period in Scottish children and that gender differences in total activity are present early in life. Tracking of total activity was only modest, but adequate assessment of tracking requires methodological research aimed at elucidating the biological meaning of accelerometer output. PMID- 12634441 TI - Associations of aging and birth cohort with body mass index in a biethnic cohort. AB - OBJECTIVE: To examine associations of aging and birth cohort with body mass index (BMI) in a biethnic cohort. RESEARCH METHODS AND PROCEDURES: This was a longitudinal closed cohort study of 14,500 white and African-American men and women, 45 to 64 years of age, followed for 9 years. Aging was defined as the length of the interval in years between baseline and following visits. Birth cohort was defined by the year in which participants were born. Mixed model analyses were used to examine associations of aging, birth cohort, and BMI in four ethnicity-gender groups. RESULTS: We found that aging was associated with an increase in BMI in white and African-American men and women. The associations between aging and BMI were stronger in the younger birth cohorts. Except for white women, younger birth cohort was associated with a higher BMI. After adjusting for aging, birth cohort was associated with an increase in BMI of 0.1 kg/m(2) [95% confidence interval (95% CI): -0.1, 0.3] among white women. The corresponding values for African-American women, white men, and African-American men are 0.5 kg/m(2) (95% CI: 0.1, 0.9), 0.6 kg/m(2) (95% CI: 0.4, 0.8), and 0.6 kg/m(2) (95% CI: 0.2, 1.0), respectively. DISCUSSION: Our analyses show that, in all except white women, people in this age range who were born later have a higher BMI at the same attained age. In all groups, people who are born later gained more weight as they aged. In general, subjects ages 45 to 64 years gained weight as they aged 9 years. PMID- 12634442 TI - Physical fitness and physical activity in obese and nonobese Flemish youth. AB - OBJECTIVE: To assess different aspects of physical fitness and physical activity in obese and nonobese Flemish youth. RESEARCH METHODS AND PROCEDURES: A random sample of 3214 Flemish schoolchildren was selected and divided into an "obese" and "nonobese" group based on body mass index and sum of skinfolds. Physical fitness was assessed by the European physical fitness test battery. Physical activity was estimated by a modified version of the Baecke Questionnaire. RESULTS: Obese subjects had inferior performances on all tests requiring propulsion or lifting of the body mass (standing-broad jump, sit-ups, bent-arm hang, speed shuttle run, and endurance shuttle run) compared with their nonobese counterparts (p < 0.001). In contrast, the obese subjects showed greater strength on handgrip (p < 0.001). Both groups had similar levels of leisure-time physical activity; however, nonobese boys had a higher sport index than their obese counterparts (p < 0.05). DISCUSSION: Results of this study show that obese subjects had poorer performances on weight-bearing tasks, but did not have lower scores on all fitness components. To encourage adherence to physical activity in obese youth, it is important that activities are tailored to their capabilities. Results suggest that weight-bearing activities should be limited at the start of an intervention with obese participants and alternative activities that rely more on static strength used. PMID- 12634443 TI - Anthropometric cutoff points for predicting chronic diseases in the Mexican National Health Survey 2000. AB - OBJECTIVE: To determine optimum anthropometric cutoffs for predicting the likelihood ratios of type 2 diabetes mellitus (DM) and hypertension (HT) in Mexicans. RESEARCH METHODS AND PROCEDURES: Data from a randomly selected, nationally representative health survey (2000) with 11,730 men [37.4 (+/- 12.9) years] and 26,647 women [37.3(+/- 12.9) years] were assessed for values of body mass index (BMI) and waist circumference (WC) for predicting DM or HT by receiver operating characteristic curve analyses. Likelihood ratios for DM and HT were calculated, and BMIs or WCs for public-health screening were developed. Subanalyses included regional data. RESULTS: Likelihood ratios of DM and HT increased from BMI values of 22 to 24 kg/m(2) in both sexes and with WC values of 75 to 80 cm in men and 70 to 80 cm in women. The best BMI cutoffs for predicting DM were 26.3 to 27.4 kg/m(2) in men and 27.7 to 28.9 kg/m(2) in women, with similar values for HT, i.e., 26.2 to 27.0 kg/m(2) and 27.7 to 28.5 kg/m(2), for men and women, respectively; WC cutoffs for DM were 93 to 98 cm in men and 94 to 99 cm in women, and cutoffs for HT were 92 to 96 cm and 93 to 96 cm for men and women, respectively. The WC cutoffs had higher sensitivity and specificity than those of BMI. DISCUSSION: The risk for DM and HT starts at lower levels of BMI and WC than those suggested by WHO. WC is a better discriminator than BMI measures for use in public health. PMID- 12634444 TI - Getting worse: the stigmatization of obese children. AB - OBJECTIVE: The prevalence of childhood obesity more than doubled in the period from 1961 to 2001. We replicated a 1961 study of stigma in childhood obesity to see what effect this increased prevalence has had on this stigma. RESEARCH METHODS AND PROCEDURES: Participants included 458 5th- and 6th-grade children attending upper-middle and lower-middle income U.S. public schools. Children ranked six drawings of same-sex children with obesity, various disabilities, or no disability ("healthy"), in order of how well they liked each child. RESULTS: Children in both the present and the 1961 study liked the drawing of the obese child least. The obese child was liked significantly less in the present study than in 1961 [Kruskal-Wallis H(1) = 130.53, p < 0.001]. Girls liked the obese child less than boys did [H(1) = 5.23, p < 0.02]. Children ranked the healthy child highest and significantly higher than in 1961 [H(1) = 245.40, p < 0.001]. The difference in liking between the healthy and obese child was currently 40.8% greater than in 1961. DISCUSSION: Stigmatization of obesity by children appears to have increased over the last 40 years. PMID- 12634445 TI - The relationship of rapid weight gain in infancy to obesity and skeletal maturity in childhood. AB - OBJECTIVE: Children with birth weight appropriate for gestational age (AGA) who also demonstrate rapid weight gain in infancy have a greater risk of being overweight or obese during childhood. A concurrent advancement in skeletal maturity would account for their greater size and would, therefore, not necessarily pose a threat of greater risk during adolescence and early adulthood. This study aims to determine whether children with rapid weight gain during infancy have advanced skeletal maturity during childhood. RESEARCH METHODS AND PROCEDURES: One hundred and ninety-three African children (boys = 108; girls = 85) of normal birth weight and gestational age were assessed from birth to 9 years. Body composition was assessed at 9 years of age by whole-body DXA, and skeletal maturity was assessed using the Tanner-Whitehouse II technique. Rapid weight gain in infancy was defined as a +0.67 change in weight-for-age Z-score between birth and 2 years. RESULTS: Rapid weight gain was experienced by over 20% of the sample. Children with rapid weight gain were significantly lighter at birth and significantly taller, heavier, and fatter throughout childhood. Chronological age and Tanner-Whitehouse II technique skeletal ages at 9 years were not significantly different between groups or between sexes within groups. DISCUSSION: Because AGA children with rapid weight gain have a greater risk of overweight and obesity but are not advanced in skeletal maturity, later adolescent adjustments toward average weight and fatness values are unlikely. The identification and monitoring of such children is of importance in reducing their risk of morbidity. PMID- 12634446 TI - Site-specific changes in the expression of fat-partitioning genes in weanling rats exposed to a low-protein diet in utero. AB - OBJECTIVE: Intrauterine growth restriction is associated with increased prevalence of the metabolic syndrome in adult life, including increased adiposity. The aim of this study was to investigate if maternal protein energy malnutrition is associated with changes in expression of genes involved in fat partitioning in weanling rats. RESEARCH METHODS AND PROCEDURES: Time-mated mothers were placed on one of two isocaloric diets, low protein [(LP), 8% protein] or control (20% protein). All mothers remained on the diet throughout pregnancy and lactation. A third group received control for 2 weeks and was switched to LP for the last week of pregnancy and lactation [late low protein (LLP) group]. Offspring were analyzed at weaning for serum glucose, nonesterified fatty acids, triglyceride, and insulin. Expression of the genes acetyl-coenzyme A carboxylase, fatty acid synthase, and carnitine palmitoyl transferase 1 were measured in liver, quadriceps muscle, and subcutaneous white adipose tissue using semiquantitative reverse transcription-polymerase chain reaction. RESULTS: LLP and LP offspring were shorter, weighed less, had reduced serum insulin and nonesterified fatty acids, and had increased serum glucose, serum triglycerides, and hepatic triglycerides. Hepatic gene expression of acetyl-coenzyme A carboxylase and fatty acid synthase was increased 2-fold in LLP and LP offspring (p < 0.001). These changes were not seen in muscle or subcutaneous white adipose tissue. CPT-1 gene expression was unaltered in all tissues examined. DISCUSSION: Maternal protein energy malnutrition programs gene expression of lipogenic enzymes in the liver of weanling offspring in a manner favoring fat synthesis that may predispose these offspring to fat accumulation and insulin resistance later in life. PMID- 12634447 TI - Subdivision of the subcutaneous adipose tissue compartment and lipid-lipoprotein levels in women. AB - OBJECTIVE: The aim of this study was to compare the relative importance of computed tomography-measured abdominal fat compartment areas, including adipose tissue located posterior to the subcutaneous Fascia, in predicting plasma lipid lipoprotein alterations. RESEARCH METHODS AND PROCEDURES: Areas of visceral as well as subcutaneous deep and superficial abdominal adipose tissue were measured by computed tomography in a sample of 66 healthy women, ages 37 to 60 years, for whom a detailed lipid-lipoprotein profile was available. RESULTS: Strong significant associations were observed between visceral adipose tissue area and most variables of the lipid-lipoprotein profile (r = -0.25, p < 0.05 to 0.62, p < 0.0001). Measures of hepatic lipoprotein synthesis such as very-low-density lipoprotein-triglyceride and cholesterol content as well as total and very-low density lipoprotein-apolipoprotein B levels were also strongly associated with visceral adipose tissue area (r = 0.57, 0.57, 0.61, and 0.62, respectively, p < 0.0001). Significant associations were found between these variables and the deep subcutaneous adipose tissue area or DXA-measured total body fat mass. However, the correlation coefficients were of lower magnitude compared to those with visceral adipose tissue area. Multivariate regression analyses demonstrated that visceral adipose tissue area was the strongest predictor of lipid-lipoprotein profile variables (7% to 48% explained variance, 0.02 > or = p < or = 0.0001). DISCUSSION: Although previous studies have generated controversial data as to which abdominal adipose tissue compartment was more closely associated with insulin resistance, our results suggest that visceral adipose tissue area is a stronger correlate of other obesity-related outcomes such as lipid-lipoprotein alterations. PMID- 12634448 TI - The estrogen antagonist EM-652 and dehydroepiandrosterone prevent diet- and ovariectomy-induced obesity. AB - OBJECTIVE: EM-652 is a pure antiestrogen in human breast and uterine cancer cells that also reduces bone loss and plasma lipid levels in the rat. This study aimed to assess the ability of EM-652, alone or with dehydroepiandrosterone (DHEA), to prevent obesity and related metabolic abnormalities induced by an obesity promoting diet and ovariectomy. RESEARCH METHODS AND PROCEDURES: Female rats were fed a high-sucrose, high-fat (HSHF) diet, were left intact or ovariectomized (OVX), and were treated with EM-652, DHEA, or both for 20 days. Variables of energy balance and determinants of lipid metabolism and insulin sensitivity were assessed. RESULTS: The HSHF diet (vs. chow) and OVX both increased energy intake and gain, as well as energetic efficiency. Both EM-652 and DHEA prevented diet- and OVX-induced energy gain mainly by decreasing fat deposition, without being additive. The modest EM-652-induced increase in liver triglycerides of intact rats was prevented by its combination with DHEA. EM-652, but not DHEA, decreased cholesterolemia. The HSHF diet and OVX reduced insulin sensitivity, an effect that was attenuated by EM-652 and abrogated by DHEA and EM-652+DHEA. Treatment with EM-652, DHEA, or their combination abolished the diet- and OVX-induced increase in adipose lipoprotein lipase activity that accompanied fat gain. DISCUSSION: EM-652 is an effective agent to prevent diet- and OVX-induced obesity and its associated cardiovascular risk factors such as insulin resistance. The addition of DHEA prevents hepatic lipid accumulation and further ameliorates insulin sensitivity. The beneficial metabolic effects of such combined steroid therapy may, therefore, eventually prove to be clinically relevant. PMID- 12634450 TI - Suspected involvement of the X chromosome in placental mesenchymal dysplasia. AB - So far, 46 cases of placental mesenchymal dysplasia have been reported worldwide. We encountered 15 cases of placental mesenchymal dysplasia (PMD) including 7 cases delivered in our hospital. The incidence of PMD in our hospital was therefore, 7/30,758 (0.02%). The PMD had a peculiar appearance. In the gross findings, large placenta with intestine-like dilatation of the vessels on the fetal side was reported. Microscopically, cistern-like dilatation of the stem villi, fetal artery thrombosis, and villous hemorrhage were reported. However, we believe most of these findings are secondary rather than the primary of mesenchymal dysplasia. Therefore, we investigated 15 other cases of mesenchymal dysplasia, and found including vascular abnormality of the stem, intermediate and terminal villi in all case of PMD. The abnormality was observed in the vessels of the periphery of the stem villi and their vessel walls were thin and appeared weak. The intermediate villous vessels were unusual, tangled. The terminal villous abnormalities showed chorangiosis and stromal hyperplasia. These findings are mesenchymal dysplasia origin. Moreover, PMD showed female-predominant. 14/15 was female among our cases, We discuss the relationship between mesenchymal dysplasia and the X chromosome in this paper. PMID- 12634449 TI - Antiandrogenic effects of dibutyl phthalate and its metabolite, monobutyl phthalate, in rats. AB - Developmental toxicity following administration of dibutyl phthalate (DBP) and its major metabolite, monobutyl phthalate (MBuP), by gavage was determined in Wistar rats. DBP on days 0-8 of pregnancy induced an increase in the incidence of preimplantation loss at 1250 mg/kg and higher and postimplantation loss at 750 mg/kg and higher. MBuP on days 0-8 of pregnancy produced an increase in the incidence of pre- and postimplantation loss at 1000 mg/kg. DBP on days 7-15 of pregnancy caused an increase in the incidence of fetuses with malformations at 750 mg/kg. MBuP on days 7-15 of pregnancy produced an increased incidence of fetuses with malformations at 500 mg/kg and higher. DBP on days 15-17 of pregnancy resulted in a decrease in the anogenital distance (AGD) of male fetuses and increase in the incidence of fetuses with undescended testes at 500 mg/kg and higher. MBuP on days 15-17 of pregnancy caused a decreased male AGD and increased incidence of fetuses with undescended testes at 250 mg/kg and higher. No effect of DBP and MBuP on the AGD was found in female offspring. The spectrum of fetal malformations, dependence of gestational days of treatment on the manifestation of teratogenicity, and alterations in development of the male reproductive system observed after administration of DBP were in good agreement with those observed after administration of MBuP. These findings suggest that MBuP may be responsible for the induction of developmental toxic effects of DBP. The doses that produced a decrease in the AGD and undescended testes in male offspring were lower than those producing maternal toxicity, fetal malformations after administration during major organogenesis, and embryonic loss. The male reproductive system may be more susceptible than other organ systems to DBP and MBuP toxicity after maternal exposure. PMID- 12634451 TI - Integration of a transposon into the Gli3 gene in the Pdn mouse. AB - The phenotype of the genetic polydactyly/ arhinencephaly mouse (Pdn/Pdn) is similar to Greig cephalopolysyndactyly syndrome (GCPS), whose responsible gene is GLI3. Suppression of Gli3 gene expression has been observed in the Pdn/Pdn and integration of retrotransposon in Gli3 gene in the Pdn mouse has been reported. Thus, the responsible gene for Pdn/Pdn is thought to be Gli3, but the site of mutation within the gene has not been demarcated. In the present study, we demonstrated that 5442 bp of early retrotransposon was inserted into intron 3 of Gli3 gene in the Pdn mouse (Gli3 (Pdn) ). This transposon had almost the same sequence as MMY17106 (EMBL). It had 317-bp long terminal repeat at both ends followed by the identical 6-bp target duplication sequence, GAGACT. Forward and reverse PCR primers were constructed in intron 3 near the insertion point, and a forward primer in the transposon was also constructed. These primers allowed us to discriminate +/+, Pdn/+ and Pdn/Pdn embryos by the PCR products. Morphological determination of the genotypes in the Pdn mouse embryos is impossible before day 12 of gestation. Quick discrimination method of genotypes developed in the present study allows us to investigate the early dysmorphogenetic mechanisms in the brain and limbs in the Pdn/Pdn embryos. Then, the dysmorphogenetic mechanisms in the Pdn/Pdn may be extrapolated to those in GCPS. PMID- 12634452 TI - An experimental model using guinea pigs to reduce accumulated dioxins in the body. AB - Dioxins are one of the accumulative endocrine disruptors that are detected from the human body. They are known to induce a wide range of adverse effects, however there are only a few methods that have been reported to reduce them. Consequently, a new effective method to reduce the dioxins in our body needs to be developed. To develop such a method, animal models are needed to be experimented on in order to evaluate each method required. In this study, an animal model using guinea pigs was made to evaluate the methods. Three weeks after the administration of 0.05 microg/kg/day of TCDD for 5 days, the serum TCDD level in the guinea pigs was almost equal to the relatively high dioxin level among human serum. Our present study suggests that the guinea pig model used in this research can be effective for further study in the method to reduce accumulated dioxins in the body. PMID- 12634453 TI - Observation of X-Y chromosomal configuration in spermatocytes of infertile PD strain male rats. AB - PD strain male rats that carry an autosomal recessive gene, preaxial duplication (gene symbol: pd), are sterile in the homozygous condition (pd/pd) due to a spermatogenic breakdown in the process of spermatogenesis at the spermatocyte and/or spermatid stage(s), although heterozygotes (pd/+) are normal. In this study, pd/pd males were examined for the presence of abnormal association of the sex chromosomes that might lead to spermatogenic breakdown. Light and electron microscopic observations of the chromosomes at meiotic prophase and metaphase in primary spermatocytes revealed several types of abnormal X-Y association and configurations in pd/pd males. However, the incidences of the abnormal configuration were comparable to those in pd/+ males. These results suggest that abnormal X-Y chromosome association in the germ cells is not a significant cause of spermatogenic breakdown in pd/pd males. PMID- 12634454 TI - Micro-lesions in Reissner's membrane evoked by acute hydrops. AB - In some pathological conditions excessive amounts of endolymph can accumulate and cause swelling, hydrops, of the membranous labyrinth. Reissner's membrane in the cochlea will distend and may even rupture. We have studied the effects of acute hydrops, followed for up to 5-6 h, in a preparation that allows continuous monitoring of structural alterations in individual cells in Reissner's membrane. This is accomplished by using laser confocal microscopy on the membrane visualized by labeling its cells with fluorescent dyes. In specimens subjected to hydrops it was observed that discrete structural defects developed in Reissner's membrane. These were seen as lesions in single cells or in groups of cells in the epithelial layer. It is suggested that through these micro-lesions the electro chemical environment of the organ of Corti can be altered causing hearing loss and tinnitus during hydrops. PMID- 12634455 TI - A glucocorticoid reduces adverse effects of adenovirus vectors in the cochlea. AB - Gene transfer using a recombinant adenovirus is a powerful tool for research and clinical applications, but its cytotoxicity and immune response limit its use, especially when repeated application of the vector is necessary. This study investigated the effects of dexamethasone (DEX)-induced immunosuppression on the outcome of adenovirus gene transfer in guinea pig inner ears. Animals received DEX for 29 days. Their inner ear was inoculated with 5 micro l of adenovirus vector twice, on days 5 and 26. Auditory brainstem response was measured on days 1, 8 and 29. The animals were sacrificed on day 29, and reporter gene expression was evaluated. In control animals that received no DEX, postinoculation threshold shifts and lesions in the organ of Corti were observed and reporter gene expression was absent. In contrast, DEX-treated ears were largely protected, and transduction of inner ear cells was readily apparent. These data demonstrate that immunosuppressive treatment can reduce the negative consequences of repeated adenovirus-mediated gene therapy. PMID- 12634456 TI - Synaptophysin immunohistochemistry during vestibular hair cell recovery after gentamicin treatment. AB - In the present study, morphometric and immunohistochemical techniques were used to evaluate the degree of synaptic recovery in the chinchilla crista sensory epithelia during various post-gentamicin-treatment periods of hair cell loss and recovery. For this purpose, two groups of animals were treated with Gelfoam pellets impregnated with 50 micro g of gentamicin implanted in the perilymphatic space within the otic capsule of the superior semicircular canal. Animals were sacrificed 1, 2 and 4 weeks after treatment. The degree of synaptic reinnervation was evaluated in the horizontal crista of the first group of animals using immunohistochemical techniques and antibodies against synaptophysin, a marker for synaptic reinnervation and synaptogenesis. Quantification of immunoreactivity in this group was made in the mid-region of the crista using the NIH 'Image' program. The second group of animals was used for quantification of the number of hair cells and supporting cells in the horizontal crista. In the normal sensory epithelium, synaptophysin immunoreactivity was found in the areas corresponding to the known distribution of afferent and efferent nerve terminals. Immunoreactivity was predominantly located within the afferent calyces of type I hair cells. No immunoreactivity was found in the supporting cells. Seven days after treatment there was a significant loss of hair cells and synaptophysin stained area (SSA). In the mid-region of the crista the loss of synaptophysin immunoreactivity was quantitatively the greatest within the central zone of this region (93%) while the loss of hair cells was the smallest. These results suggest that afferent and efferent nerve terminals were also severely affected by the ototoxic treatment. Four weeks after treatment corresponding to the end of the recovery phase of gentamicin ototoxicity, there was a proportional increase in the number of hair cells and of the degree of SSA in the mid-region of the crista. The number of hair cells recovered to 58% with a recovery of SSA to 54% of normal. These results suggest that a greater fraction of synaptophysin expression within the sensory epithelium depends on the presence of afferent calyceal endings, which are greatly affected by gentamicin. Also, these results demonstrate a significant level of reinnervation of the newly regenerated hair cells, forecasting the potential for functionality of the regenerated hair cells. PMID- 12634457 TI - Theoretical models for the mechanisms of benign paroxysmal positional vertigo. AB - Benign paroxysmal positional vertigo (BPPV) is a common vestibular disorder resulting from a malfunction of the semicircular canal. Prior studies attempting to elucidate the mechanics of BPPV have focused on clinical and laboratory findings, and have discussed theoretical aspects only in qualitative terms. The goal of this study is to create a mathematical description of BPPV mechanics based on biophysical principles, in order to improve the physiopathological understanding of the most common varieties of the disorder, canalithiasis and cupulolithiasis. First, the abnormal stimulus being applied to the vestibular system during BPPV episodes is estimated by an analysis of the vestibulo-ocular reflex (VOR) response of clinical BPPV patients. The magnitude and time course of the head motion stimulus that would, in a normal test subject, produce a VOR response similar to that observed during BPPV nystagmus are inferred from eye motion data using an inverse transfer function approach. Next, physicomathematical models to describe the mechanics of the abnormal stimulations of the crista are created: the cupulolithiasis model is based on the principle that dense particulate matter becomes attached to the cupula of affected patients and effectively changes the specific gravity of the cupula; the canalithiasis model is based on the hydrodynamic effects from a cluster of particles falling within the canal endolymph. The stimuli predicted by these two models are compared to the stimuli estimated from physiological and clinical studies. The model predictions are found to be consistent with the empirical evidence for a reasonable set of model parameters. The cupulolithiasis model predicts a value of 0.69 micro g as the mass of particles contributing to the disorder. The canalithiasis model predicts a value of 0.087 micro g. These results support and expand our understanding of the mechanisms underlying the production of the cupulolithiasis and canalithiasis varieties of vertigo in humans. On the basis of these models, several predictions that can help the clinician improve the evaluation of BPPV patients are discussed. PMID- 12634458 TI - The neonate has a temporary conductive hearing loss due to fluid in the middle ear. AB - Postnatal functional changes in the activity of the ear and auditory pathway in neonatal guinea pigs [from day of birth (postnatal day, PND = 0), PNDs 1-4, 7 and then weekly up to 7 weeks] were studied as a model of maturation of hearing in human neonates. On the day of birth there were signs of a conductive hearing loss: negative middle ear pressure, auditory nerve brainstem evoked response (ABR) threshold elevation, ABR wave 1 latency prolongation and low amplitude otoacoustic emissions. The conductive hearing loss is probably a result of the (amniotic) fluid found in the neonatal middle-ear cavity. Over the next PNDs, this conductive hearing loss was resolved. In order to confirm this neonatal conductive hearing loss and its resolution, saline was instilled into the middle ear of guinea pigs. This induced signs of a conductive hearing loss similar to those seen in the neonatal guinea pigs which disappeared with clearance of this fluid. Therefore it may be concluded that most of the changes in auditory function seen over the first PNDs are due to absorption of amniotic fluid from the middle-ear cavity. PMID- 12634459 TI - Central hearing loss with a bilateral inferior colliculus lesion. AB - A case is presented of a 43-year-old man with bilateral lesions of the inferior colliculus (IC) due to central nervous system lymphoma. Our patient was found to have relatively normal pure-tone averages with severely reduced bilateral word recognition scores. This case illustrates that the presentation of a partial bilateral IC lesion may be of severely reduced word recognition accompanied by nearly normal pure-tone hearing. PMID- 12634460 TI - Air travel-associated venous thromboembolism. AB - Long-distance air travel is increasing and cases of venous thromboembolism (VTE) following air travel have attracted both considerable public attention and legal claims against airlines. VTE is a common disorder worldwide with a notably high incidence in older individuals. Many biochemical factors that lead to, or accentuate, thrombus formation are associated with increased risk of VTE. These factors include thrombophilia, activated protein C resistance and factor V Leiden, prothrombin gene mutation, antiphospholipid antibodies, protein S and protein C deficiencies, and methylene tetrahydrofolate reductase polymorphism and homocysteinemia. Individual physical characteristics including age, weight and height are significant for personal risk of VTE as are other factors such as use of oral contraceptives in women. In the case of air travel-related venous thrombosis, superimposed upon these individual factors are the environmental factors directly related to air travel. Travel-related factors include stasis associated with prolonged periods of immobility, physiological stresses resulting from exposure to the cabin environment (low humidity and hypoxia) in long-haul flight and other in-flight factors. It is suggested that passenger behavior (movement, avoidance of dehydration and of alcohol) and appropriate pharmacological prophylaxis for high-risk travelers can reduce the likelihood of VTE. Physical prophylaxis (use of compression stockings or in-flight exercise devices) may also be of general benefit to passengers. It is recommended that airlines become more proactive in educating passengers concerning the dangers of VTE and in promoting passenger actions that can reduce risk. Airlines should also work to avoid cramped seating conditions (seat size and pitch) that contribute to prolonged immobility. Governments and regulatory authorities should mandate the provision of adequate seating conditions and a good cabin environment and should support studies that will define risks and determine the efficacy of protocols to minimize dangers of VTE. Increased long-haul air traffic and an aging population suggest that travel-related VTE may present a growing healthcare threat and has highlighted a need for additional biomedical research into the causes and potential solutions to this problem. PMID- 12634461 TI - Comparative evaluation of anoxomat and conventional anaerobic GasPak jar systems for the isolation of anaerobic bacteria. AB - OBJECTIVES: To evaluate the performance of the Anoxomat, in comparison with the conventional anaerobic GasPak jar system, for the isolation of obligate anaerobes. METHOD: Anoxomat, model WS800, and anaerobic GasPak jar system (Oxoid) were evaluated. Anoxomat system utilized a gas mixture of 80% N(2), 10% CO(2) and 10% H(2), while the GasPak used a gas mixture of 90% H(2) and 10% CO(2). An anaerobic indicator within the jars monitored anaerobiosis. A total of 227 obligate anaerobic bacteria comprising 116 stock strains, 5 ATCC reference strains and 106 fresh strains, representing different genera, were investigated for growth on anaerobic agar plates and scored for density, colony sizes, susceptibility zones of antibiotic inhibition and the speed of anaerobiosis (reducing the indicator). RESULTS: The results demonstrate that the growth of anaerobic bacteria is faster inside the Anoxomat jar than in the anaerobic GasPak jar system. Of the 227 strains tested, the colonies of 152 (67%) were larger (by size range of 0.2-2.4 mm) in the Anoxomat at 48 h than in the GasPak jar compared with only 21% (range 0.1-0.3 mm) that were larger in the GasPak than in the Anoxomat. The remaining 12% were equal in their sizes. There was no measurable difference in the colony sizes of the reference strains. The Porphyromonas asaccharolytica strains failed to grow within the GasPak system but grew inside the Anoxomat. With the Anoxomat, anaerobiosis was achieved about 35 min faster than in the GasPak system. The density of growth recorded for 177 (78%) strains was heavier in the Anoxomat than in the GasPak jar. The zones of inhibition of the antibiotics tested were not different in the two systems. CONCLUSION: The Anoxomat system provided superior growth, in terms of density and colony size, and achieved anaerobiosis more rapidly. Evidently, the Anoxomat method is more reliable and appears to support the growth of strict anaerobes better. PMID- 12634462 TI - Diabetes mellitus and lung function. AB - OBJECTIVES: To assess the nature of pulmonary dysfunction in type 1 diabetes and the relationship of pulmonary function tests to diabetic factors and complication. SUBJECTS AND METHODS: Sixteen type 1 diabetic patients and 26 control subjects matched for age and sex were studied. We performed spirometry measurements and measured pulmonary diffusing capacity (DL(CO)) in sitting and supine position by the single-breath method corrected by alveolar volume (VA). Glycosylated hemoglobin (HbA(Ic)), retinopathy and microalbuminuria were included as parameters of metabolic control and diabetic complications. RESULTS: Diabetic patients showed a significant reduction of the following pulmonary function tests (% predicted value) as compared with control subjects: total lung capacity (TLC, 92.6 +/- 14.5 vs. 113.9 +/- 17.5, p < 0.001), lung diffusing capacity in sitting position (DL(CO), 90.4 +/- 21.1 vs. 107.7 +/- 15.6, p = 0.004), lung diffusing capacity in supine position (DL(CO), 88.3 +/- 19.3 vs. 111.9 +/- 19.9, p = 0.001). The differences in diffusing capacity corrected by alveolar volume in sitting and supine position (DL(CO)/VA) were not significant. By changing the posture from sitting to supine position both diabetic patients and control subjects significantly increased DL(CO)/VA (103.4 +/- 17.7 vs. 112.7 +/- 22.3, p = 0.046 and 99.5 +/- 13.4 vs. 114.4 +/- 13, p < 0.001, respectively). There was no correlation between pulmonary function tests and diabetic complications. CONCLUSION: These data indicate that type 1 diabetic patients showed reduced TLC and DL(CO), features of pulmonary restrictive dysfunction. There was no correlation between abnormal pulmonary function and the presence of other diabetic complications. PMID- 12634463 TI - Surgical treatment of thoracic and lumbar tuberculosis by anterior interbody fusion and posterior instrumentation. AB - OBJECTIVE: To evaluate the efficacy and clinical outcome of simultaneous or sequential anterior and posterior surgical approaches in the management of spinal tuberculosis in the form of anterior extirpation of the tuberculous lesion, strut bone grafting of the defect produced and posterior instrumentation for spinal fixation. SUBJECTS AND METHODS: Twenty-two patients who had tuberculosis of the thoracic and lumbar spine with moderate to severe localized kyphosis and variable degrees of neurological deficits were treated at Al Razi Hospital (Kuwait) in the period from 1998 to 2000 by anterior debridement and autogenous strut bone grafting with simultaneous or staged posterior spinal fixation using either USS or SOCON spinal instrumentation. Appropriate antituberculosis treatment was given to all patients for 9-12 months. The postoperative follow-up period was 18 months. RESULTS: Of the 22 cases the average of preoperative kyphosis was 42 degrees. The average of immediate postoperative correction was 27 degrees. At the last follow-up the average correction was 24 degrees and the loss of correction did not exceed 3 degrees. Average fusion times were 5 months for one-segment fusions and 8 months for two-segment fusions. There was no recurrence of the disease in any of the cases. CONCLUSION: Posterior instrumental stabilization and anterior interbody fusion were found to be effective in arresting the disease, correcting kyphotic deformity and maintaining correction until solid spinal fusion. PMID- 12634466 TI - The relationship of recurrent spontaneous miscarriage with reproductive failure. AB - OBJECTIVE: To investigate the reproductive performance of non-pregnant women with recurrent spontaneous miscarriage (RSM) and the association between RSM and polycystic ovary syndrome (PCOS) in reproductive failure. SUBJECTS AND METHODS: Fifty non-pregnant women with 3 or more consecutive RSM attending our RSM clinic were evaluated prospectively. Detailed history and physical examination were completed. Extensive laboratory investigations, ultrasonographic, radiological and specialized procedures/tests were performed. Those patients who did not achieve pregnancy within the first 6 months of follow-up had appropriate treatment options implemented. The outcome of any subsequent pregnancies was analyzed. RESULTS: The mean age of the patients was 33.8 +/- 4.6 years (range 22 43 years). The aetiological factors associated with RSM were chromosome anomaly, uterine abnormality, antiphospholipid syndrome, PCOS and infections; 40% were 'unexplained'. Thirty patients (60%) were able to achieve viable pregnancies which progressed to term, resulting in live births. Fourteen of the other 20 patients had high body mass index (BMI) >30 and evidence of PCOS. Of these 20 patients, 13 eventually achieved pregnancies which ended again in first-second trimester abortions (cause undetermined). The remaining 7 patients (4 with high BMI) who had a combination of male factor/protracted PCOS/ovarian failure did not achieve pregnancies. CONCLUSION: The association of RSM with PCOS resulted in poor reproductive performance, which may progress to reproductive failure. PMID- 12634464 TI - A comparative study of samarium-153-ethylenediaminetetramethylene phosphonic acid with pamidronate disodium in the treatment of patients with painful metastatic bone cancer. AB - OBJECTIVE: To assess the therapeutic efficacy and toxicity of samarium-153 ethylenediaminetetramethylene phosphonic acid ((153)Sm-EDTMP) and pamidronate disodium in patients with painful metastatic bone cancer. SUBJECTS AND METHODS: Eighteen patients with histopathologically confirmed malignancy and multifocal bone metastases were randomized into two equal groups of 9 patients each. Group A was treated with (153)Sm-EDTMP, while group B was treated with pamidronate disodium. The pain score for each patient was recorded before and after therapy using visual analogue scales that graded both the intensity and frequency of the bone pain. Therapeutic response was classified as inefficient, mild, effective and excellent. RESULTS: Pain score in each group prior to therapy was more than 6. In group A, 2 (22.2%) and 7 (77.8%) cases showed mild and effective response, respectively. The therapeutic efficacy of (153)Sm-EDTMP was adjudged to be 77.8%. Transient myelosuppression was generally mild and reversible with white blood cells and platelets recovering after 6 weeks. In group B, palliative response in 4 cases (44.4%) was inefficient, in 1 case (11.1%) mild, in 3 cases (33.3%) effective and in 1 case (11.1%) excellent, with a therapeutic efficacy of 44.4% for pamidronate disodium. No hematological toxicity was noted. CONCLUSION: The data showed that the therapeutic efficacy of (153)Sm-EDTPM was higher than that of pamidronate disodium (for pain relief maintained more than 3 weeks) and its incidence of blood toxicity was also higher than that of pamidronate disodium. PMID- 12634465 TI - Prevalence of caries and lesions treatable through the ART approach in high caries risk schoolchildren in Damascus, Syria. AB - OBJECTIVE: To determine the prevalence of dentinal lesions treatable through the atraumatic restorative treatment (ART) approach for high caries risk schoolchildren. SUBJECTS AND METHODS: A convenience sample was taken of 1,542 grade 1 and 931 grade 2 children from 21 primary schools in a low socio-economic area in Damascus. Dentinal lesions in which the smallest excavator (diameter of 0.9 mm) could enter the cavitated lesion were considered treatable using the ART approach. Mann-Whitney U and ANOVA tests were used to analyse the data. RESULTS: The mean dmfs and dmft scores of the 6- to 7-year-olds (grade 1) were 9.0 and 4.4, respectively. The mean DMFS and DMFT scores of the 7- to 8-year-olds (grade 2) were 1.6 and 1.4, respectively. At least 89.9 and 54.9% of the dentinal lesions in the deciduous and permanent dentition that were in need of a restoration, respectively, were diagnosed as treatable using ART. CONCLUSION: A high proportion of dentinal lesions in need of restorations in both deciduous and permanent dentitions in this high caries risk group of 6- to 8-year-olds were diagnosed as being treatable using the ART approach. PMID- 12634467 TI - Surgical treatment and follow-up of pulmonary hydatid cyst. AB - OBJECTIVE: To report our experience of surgical procedures in the management of hydatid cysts of the lung and to assess the effect of postoperative chemotherapy. SUBJECTS AND METHODS: In a longitudinal cohort study, 64 consecutive patients who presented with hydatid cysts of the lung from 1994 to 1998 were included. The main measures were: characteristics on presentation, operative techniques, postoperative morbidity, and the outcome of treatment. The mean age was 28 years (range 4-65 years). The most common symptoms were cough, chest pain, fever, and hemoptysis. Chest radiographs and computed tomograms were the main method of diagnosis. Pulmonary cystotomy and capitonnage were performed in 46 patients. Pulmonary resection was needed in 8 of 64 patients. Simultaneous combined resection of hydatid cysts through thoracotomy with transdiaphragmatic removal of liver cysts was performed in one stage in 13 patients. RESULTS: Chest radiographs and CT scans showed a smoothly outlined spherical opacity in 45 patients. Other radiographic findings included ill-defined shadow (in 8 patients), pleural effusion (7 patients), air fluid level (3 patients), and hydropneumothorax in a single patient. There were 8 (12.5%) cases of immediate postoperative complications. These occurred mostly in patients who had cysts larger than 10 cm (p = 0.003). Thirty-four patients were treated by a 3-month course of albendazole chemotherapy. Four patients (6%) had recurrences of the disease during the follow up period. These recurrences occurred in patients with large cysts >10 cm (p = 0.001). CONCLUSION: Conservative surgical methods are the preferred surgical techniques. Postoperative chemotherapy with albendazole for 3 months is recommended. PMID- 12634468 TI - Response to cisplatin-etoposide treatment and survival in patients with small cell lung cancer in North Lebanon. AB - OBJECTIVE: To study the clinical features of small-cell lung cancer (SCLC) cases diagnosed in adults in North Lebanon. PATIENTS AND METHODS: Descriptive analysis of 125 SCLC cases diagnosed in a tertiary care hospital in North Lebanon. RESULTS: One hundred and twenty-five patients diagnosed with SCLC comprised the study group; 109 male and 16 female patients aged from 35 to 84 years with a mean age of 56 +/- 7.4 years. 95.4% of male and 31.2% of female patients were current smokers. The most frequent symptoms were cough, dyspnea, and chest pain in 24.8, 18.4, and 16.8%, respectively. Staging procedures were performed in all patients: 49 (39.2%) patients had limited stage (LS) and 76 (60.8%) patients had extensive stage (ES) disease. Metastasis was most frequent to bones (40.8%) followed by liver (24.8%) and brain (14.4%). The response rates, including both complete and partial responses to therapy, were 80, 41.5, and 52% for LS, ES, and overall patients, respectively. Median survival was 311, 163, and 201 days for LS, ES, and overall patients, respectively. CONCLUSION: Clinical features of the patients were very similar to those reported in the literature except that there were lower median survival values and response rates to treatment, including both complete and partial responses for both LS and ES disease. The results should be considered preliminary with regard to the activity of the chemotherapy regimen, given the statistical limitation of the small number of patients. PMID- 12634469 TI - Female genital cutting: types, motives and perineal damage in laboring Egyptian women. AB - OBJECTIVES: To study the prevalence of female genital cutting (FGC) in a nulliparous population admitted for childbirth. In addition, the type of FGC, the motives supporting FGC and perineal damage associated with this practice were evaluated. SUBJECTS AND METHODS: A prospective cross-sectional study was conducted in the labor ward, Department of Obstetrics and Gynecology, Assiut University Hospital, Assiut, Egypt. Two hundred and fifty-four primigravid women in active labor were recruited over a period of 20 months. The subjects responded to a questionnaire and obstetric and pelvic examinations were carried out. The type of cut (circumcision) and extent of tissue removal were recorded. Data was gathered concerning possible motives for FGC, rates of episiotomy and incidence of perineal tears. RESULTS: All women recruited had been circumcised; 51% had type I cut and 49% had type II. Adherence to tradition was the most common motive for the operation (46.5%). Ninety-five percent of the study population had an episiotomy. The incidence of perineal tears was 1.6%. CONCLUSIONS: Female genital cutting (only types I and II) was confirmed in all patients in the study. There was a low incidence of perineal tears and a high episiotomy rate (95%). Episiotomy should be performed in all cases where FGC has made the vulva/vagina inelastic. PMID- 12634470 TI - Pediatric idiopathic pulmonary hemosiderosis diagnosed by sputum analysis: plain radiography and computed tomography findings. AB - OBJECTIVE: Idiopathic pulmonary hemosiderosis (IPH) is an uncommon disorder, which is characterized by recurrent hemoptysis, iron deficiency anemia and diffuse parenchymal infiltration on chest radiographs in pediatric patients. We wish to present clinical and radiological (plain radiography and CT) findings of this rare pathology. CLINICAL PRESENTATION AND INTERVENTION: A 14-year-old girl was admitted to the pediatric emergency department with complaints of cough, dyspnea, fatigue and bloody sputum for 6 months. She had been hospitalized 3 times during this period and received antibiotics and blood transfusion. Chest X rays revealed prominent perihilar and bibasilar consolidation. CT showed a ground glass pattern and consolidated areas with increased density. Sputum analysis yielded hemosiderin-laden macrophages. With presumptive diagnosis of IPH, prednisolone was administered. Her symptoms improved on the 5th day of treatment and 1 month later, plain chest radiography demonstrated marked improvement. CONCLUSION: Although IPH is a rare condition, the diagnosis of IPH should be considered, among others, in a patient with hemoptysis and bilateral infiltration in the chest X-ray. This may prevent antibiotic misuse and risk of death due to severe hemorrhage. PMID- 12634471 TI - Wegener's granulomatosis with parotid gland involvement and pneumothorax. AB - OBJECTIVE: Wegener's granulomatosis is a systemic vasculitis characterized by necrotizing granulomatous lesions mostly involving the upper and lower respiratory tract. The disease rarely causes parotid gland involvement and pneumothorax. We report a case of Wegener's granulomatosis involving parotid gland, and complicated with a pneumothorax. CLINICAL PRESENTATION: A 45-year-old man admitted with a 3-week history of painful left parotid gland enlargement and hemoptysis. On physical examination a painful and hard mass was detected on the left pre-auricular area. Cervical CT revealed a 2 x 1.5 cm hypodense lesion mimicking an abscess on the left parotid gland. Chest radiograph and thorax CT demonstrated nodular and cavitating opacities on the right and left upper zones. There were numerous erythrocytes in urine sediment. The drained pus material from the parotid abscess demonstrated only gram-positive cocci (Staphylococcus aureus). Two weeks treatment with teicoplanin resulted in no improvement. Meanwhile, parotid gland biopsy revealed necrotizing granulomatous inflammation. There was a sixfold increase in serum cANCA levels. With the diagnosis of Wegener's granulomatosis, cyclophosphamide and prednisolone were initiated. However, 1 month later, pneumothorax developed as a complication of rupture of a cavitary lesion. CONCLUSION: Parotid gland swelling may be the initial presenting symptom of Wegener's granulomatosis. It can be confused with infectious or malignant diseases of the gland, and the lung involvement may be complicated with pneumothorax. PMID- 12634472 TI - Morals and the human brain: a working model. AB - Morality has been at the center of informal talks and metaphysical discussions since the beginning of history. Recently, converging lines of evidence from evolutionary biology, neuroscience and experimental psychology have shown that morality is grounded in the brain. This article reviews the main lines of investigation indicating that moral behavior is a product of evolutionary pressures that shaped the neurobehavioral processes related to the selective perception of social cues, the experience of moral emotions and the adaptation of behavioral responses to the social milieu. These processes draw upon specific cortical-subcortical loops that organize social cognition, emotion and motivation into uniquely human forms of experience and behavior. We put forth a model of brain-behavior relationships underlying moral reasoning and emotion that accommodates the impairments of moral behavior observed in neuropsychiatric disorders. This model provides a framework for empirical testing with current methods of neurobehavioral analysis. PMID- 12634473 TI - The right supratemporal plane hears the distance of objects: neuromagnetic correlates of virtual reality. AB - The neural mechanisms of auditory distance perception, a function of great biological importance, are poorly understood. Where not overruled by conflicting factors such as echoes or visual input, sound intensity is perceived as conveying distance information. We recorded neuromagnetic responses to amplitude variations over both supratemporal planes, with and without auditory spatial simulations. In the absence of other cues for distance, including those provided by auditory virtual reality, amplitude changes elicited enhanced preattentive responses over the right temporal lobe, indicating hemispheric lateralization of the 'where' pathway in the human. Lesion studies in monkeys and humans have shown that the rostral part of the right superior temporal cortex contributes to spatial awareness in the visual domain. Our data indicate that the distance to a sound source is processed within the adjacent right auditory cortex, thus extending the recent model of a right-hemisphere temporal multisensory matrix that subserves the integration of space-related data across visual and auditory modalities. PMID- 12634474 TI - Up-regulation of striatal adenosine A(2A) receptors in schizophrenia. AB - Adenosine A (2A) receptors have been implicated in the pathophysiology of schizophrenia by clinical, anatomical, biochemical and genetic studies. We hypothesized that a genetically determined low number of adenosine A (2A) receptors could be a vulnerability factor for the development of the disease. The density of adenosine A (2A) receptors was investigated in human postmortem striatum of patients with schizophrenia (n = 9) and matched controls ( n= 9) using [ H)CGS 21680 as a radioligand probe. The maximum number of binding sites (B) (max) was 70% higher in patients with schizophrenia than in matched controls (609.4 +/- 259.1 354.0 +/- 156.4 fmol/mg protein, p=0.04). No significant difference could be discerned for the affinity of caffeine for adenosine A receptors between patients and controls. The increase in receptor density correlated with the dose of antipsychotic medication in chlorpromazine equivalents (r =0.61, = 0.014). We failed to provide evidence for a genetically determined reduction of adenosine A 2(A) receptors in schizophrenia. Instead, consistent with findings from animal experiments, our observation supports a role of adenosine A (2A) receptors in the molecular effects of antipsychotic drugs. PMID- 12634475 TI - Myelin basic protein-reactive T cells induce conduction failure in vivo but not in vitro. AB - The ability of myelin basic protein (MBP)-reactive T cells to induce conduction failure was investigated and. With the model, somatosensory evoked potentials (SEP) were recorded before and during adoptively transferred experimental autoimmune encephalomyelitis (EAE) in Lewis rats. Maximum amplitude SEP were reached within 15 min of anesthesia. During EAE, the SEP decreased considerably and their onset was delayed. However, the compound action potentials (CAPs) recorded from Lewis rat optic nerves incubated with encephalitogenic T cells were not affected, emphasizing the importance of environmental factors. This study shows that the model described here is an useful means of investigating the neurological disorders associated with EAE. PMID- 12634476 TI - Modulation of cortex-muscle oscillatory interaction by ischaemia-induced deafferentation. AB - We studied the effect of sensory feedback on the oscillatory interaction between activity of the motor cortex and the spinal motoneuron pool during isometric contraction. After inducing ischaemic sensory deafferentation in the upper limb in six subjects, we calculated coherences between simultaneously recorded whole scalp magnetoencephalographic (MEG) signals and electromyographic (EMG) signals from the first dorsal interosseus muscles. We expected that the dominant frequency of coherence would change if there were interaction through a sensory feedback loop. However, the MEG-EMG coherence frequency did not change significantly during ischaemia. The strength of the coherence was reduced during ischaemia, but returned to the pre-ischaemic level after ischaemia had ended. Reduction of sensory feedback may thus indirectly reduce the amount of corticomuscular coherence, but the lack of change in the dominant coherent frequency suggests that a sensory feedback loop is not essential for the generation of this coherence. PMID- 12634477 TI - Brain regions showing increased activation by threat-related words in panic disorder. AB - Threat-related stimuli consistently activate the posterior cingulate cortex in normal subjects and have exaggerated effects on memory in patients with panic disorder. We hypothesized that panic patients would show increased response to threat-related stimuli in the posterior cingulate cortex. While undergoing fMRI, six panic patients and eight healthy volunteers made valence judgements of threat related and neutral words. Both groups showed threat-related activation in the left posterior cingulate and left middle frontal cortices, but the activation was significantly greater in panic patients. Panic patients also had more right>left asymmetry of activation in the mid-parahippocampal region. The increased responsivity observed in the posterior cingulate and dorsolateral prefrontal cortices is consistent with the hypothesis that panic disorder patients engage in more extensive memory processing of threat-related stimuli. PMID- 12634478 TI - Change in endothelial nitric oxide synthase in the rat retina following transient ischemia. AB - Patterns of endothelial nitric oxide synthase (eNOS) expression in retinal ischemia were studied utilizing a transient high intraocular pressure (HIOP) model. We investigated neuronal cell damage and changes in eNOS immunoreactive expression in the ischemic retina, and its relationship to the neuroprotection of betaxolol treatment after ischemic injury. Immunohistochemical staining for eNOS was performed at 3, 7, 14 and 28 days after ischemia/reperfusion. In controls, eNOS immunoreactivity was detected in retinal vessels, but was not detected in neurons. After ischemia/reperfusion, the intensity of eNOS immunoreactivity increased in both retinal vessels and the ganglion cell layer (GCL) compared with controls. eNOS-positive neurons were induced first in the inner nuclear layer (INL) 7 days after reperfusion. However, when experiments were carried out on animals that had been treated with betaxolol after ischemia/reperfusion, the intensity of eNOS immunoreactivity decreased compared to the untreated ischemic retinas. These results suggest that an increase in eNOS expression could be associated with the degenerative changes in the ischemic retina, and that betaxolol treatment appears to protect retinal tissue from ischemic damage. PMID- 12634479 TI - Selective caudate atrophy in multiple sclerosis: a 3D MRI parcellation study. AB - Deep gray matter damage may be an important component of the multiple sclerosis (MS) disease process. We tested whether caudate atrophy occurs in MS, and whether it correlates with conventional MRI or clinical markers of disease progression. Caudate nuclei of 24 MS patients and 10 age-matched healthy controls were traced, normalized, reconstructed and visualized from MRI scans. Normalized bicaudate volume was 19% lower in MS controls ( p< 0.001), an effect that persisted after adjusting for whole-brain atrophy ( p< 0.008). Caudate volume did not correlate with disease duration, physical disability score, whole-brain atrophy, or total T2 hyperintense or T1 hypointense lesion load (all p > 0.05). We conclude that selective caudate atrophy is associated with MS and may occur through direct mechanisms. PMID- 12634480 TI - Reduced programmed cell death in brains of serotonin transporter knockout mice. AB - Serotonin (5-HT) is known to reduce apoptosis and in rodent models of brain ischemia. Modulation of programmed cell death during neural development was assessed in early postnatal brains of serotonin transporter (5-HTT) knockout mice, characterized by elevated extracellular 5-HT levels. The number of apoptotic cells visualized at postnatal day-1 (P1) by ISEL+ or TUNEL staining was significantly reduced in the striatum, thalamus/hypothalamus, cerebral cortex and hippocampus of 5-HTT knockout mice, compared to wild type and heterozygote mice, with differences displaying an increasing fronto-caudal gradient and regional specificity. These findings underscore 5-HT roles in the regulation of programmed cell death during brain development, and spur interest into pharmacological interventions aimed at relieving pathological apoptosis by potentiating serotoninergic neurotransmission. PMID- 12634481 TI - Childhood and current cognitive function in healthy 80-year-olds: a DT-MRI study. AB - Diffusion tensor MR imaging (DT-MRI) yields information on early pathological changes in white matter of the ageing brain which may correlate with cognitive function. However, because individuals vary in their cognitive ability, a measurement of prior cognition from youth is required to understand fully the significance of MR imaging changes associated with ageing. Here, diffusion tensor parameters and cognitive function were measured in a cohort of 30 older subjects whose cognitive ability was measured at age 11 and 80. There was a significant correlation between diffusion anisotropy measured in the centrum semiovale at age 80 and mental ability determined at age 11 and 80. These novel results suggest, that MR imaging studies of white matter structure and its relationship to mental ability in ageing should control for early life cognition. PMID- 12634482 TI - cDNA encodes Xenopus P2Y(1) nucleotide receptor: expression at the neuromuscular junctions. AB - A cDNA encoding P2Y(1) receptor was isolated by cross-hybridization with chicken homolog. The deduced amino acid sequence of P2Y(1) receptor with 361 amino residues is 80-85% identical to human, rodent and avian homologs. When the cDNA was expressed in mammalian cells, the activation of P2Y(1) receptor by adenine nucleotides stimulated the accumulation of inositol phosphate, and adenosine 3',5'-bismonophosphate (A3P5P) or other antagonists blocked its action; these pharmacological properties showed resemblance of P2Y(1) receptor family in higher vertebrate. A transcript encoding P2Y(1) receptor at approximately 3.2 kb was revealed in the brain, spinal cord and muscle of adult, and it is strongly expressed in developing brain, spinal cord and myotomal muscles of the embryos by hybridization. P2Y(1) receptor was shown to be restricted to the neuromuscular junctions and co-localized with AChRs in adult muscle. These results support the notion that ATP and its P2Y(1) receptor subtype are effectors in organizing the post-synaptic apparatus. PMID- 12634484 TI - NADPH-diaphorase-positive neurons in the auditory cortex of young and old rats. AB - Age-related changes in NADPH-diaphorase (NADPH-d)-positive neurons were examined in the auditory cortex of young (3 months old) and very old (36 months old) rats (strain Long Evans). In very old rats a significant reduction was found in the thickness of the auditory cortex, to 54% of that in young animals, as well as changes in the shape and configuration of nerve cell bodies and dendrites. Quantitative analysis demonstrated an age-related increase in the number of dendritic segments and dendritic branching points. The length of dendrites in NADPH-d-positive neurons and their density increased in very old rats. The total number of NADPH-d-positive neurons within the Te 1 and Te 3 fields was 13% lower in the old rats than in the young. PMID- 12634485 TI - A neural model for visual selection of grouped spatial arrays. AB - Psychophysical and electrophysiological studies indicate that visual attention operates on early retinotopic maps and selects spatially grouped arrays of locations which correspond to objects or perceptual groups. A neural model is proposed which is able to select an array of locations labelled by the same activity level and suppress all other regions that are not in the focus of attention. The model uses self-recurrent dendritic inhibition to compute the maximum activity level in the input and suppress a feedforward flow of activity from input to selection layer at unattended locations. Computer simulations also illustrate the model's ability to detect abrupt onsets of new objects in the visual scene, perform visual search and track moving objects. PMID- 12634483 TI - Galanin expression in sensory neurons after nerve compression or transection. AB - Galanin is probably involved in nociceptive sensory processing in spinal cord. We investigated whether a common injury, peripheral nerve compression, induced up regulation of galanin (immunocytochemistry) in sensory neurons in rats 6 or 14 days post-injury and compared the response with other nerve injuries. Sciatic nerve compression increased the number of galanin positive sensory neurons as compared to uninjured and contralateral dorsal root ganglia. Complete transection was more efficient than a partial transection and a slight compression injury as an inducer of galanin. Mainly small diameter sensory neurons became positive but also some large diameter neurons. We conclude that nerve compression up-regulates galanin in sensory neurons. The extent of the induction could be related to the severity of nerve injury. PMID- 12634486 TI - Memory impairment in humans after bilateral damage to lateral temporal neocortex. AB - The hippocampus and its adjacent medial temporal regions are crucial for episodic memory. However, it is far less obvious whether other temporal regions beyond the medial part are important for memory processing. The memory performance of a group of patients with bilateral lesions over the lateral temporal lobe sparing the hippocampus was assessed and compared with that of patients with relatively spared cortex and patients with bilateral damage to the lateral temporal lobe and the hippocampus. The results demonstrate that bilateral damage to the lateral temporal lobe results in memory impairment across verbal and visual modalities. PMID- 12634487 TI - Premovement gating of somatosensory evoked potentials after tibial nerve stimulation. AB - Somatosensory evoked potentials (SEPs) are attenuated or gated during movement. The mechanism for this includes both centrifugal gating of afferent input and competition with other afferents caused by the movement (peripheral gating). Using a paradigm in which the signal for triggering movement is the electric stimulus for SEPs, we studied the gating of SEPs after tibial nerve stimulation prior to foot movement, and compared it with that during counting task. Significant gating was found for P40 component, which distributed centrally and ipsilaterally to the side of the stimulation, whereas the contralateral N40 component showed no changes. Dissociated gating of P40 and N40 indicates multiple generators of these components, in contrast to the previous view of a single generator dipole projecting tangentially. Together with the previous findings in median SEPs, these gating phenomena should represent a general mechanism for sensori-motor integration in preparation for limb movement. PMID- 12634488 TI - Alpha and beta oscillatory changes during stimulus-induced movement paradigms: effect of stimulus predictability. AB - We studied the effect of stimulus predictability on the alpha and beta changes observed in central regions during stimulus-induced movement paradigms. Six young volunteers were instructed to extend briskly their dominant wrist as soon as possible after hearing a 2000 Hz sound. Two sequences of stimuli were presented to each subject, the first rhythmic at 1/6 s and the second with random intervals between 5 and 13s. A time-frequency analysis of nonphase-locked activity in the 7 37 Hz range was performed on stimulus-centred EEG sweeps using wavelet filters and Gabor transforms. During the sequence of predictable rhythmic stimuli, stimulus-induced movements were accompanied by a decrease in beta activity that began contralaterally about 1 s prior to the stimulus and extended to both sides later on. This decrease was followed by a rebound after the end of the movement. In the alpha band, the decrease observed started just after the sound. During the sequence of non-predictable, random stimuli, stimulus-induced movements were accompanied by a shorter and smaller alpha and beta-ERD, that started after the stimulus. The presence of a pre-stimulus beta ERD only in the rhythmic predictable paradigm suggests that central pre-movement beta ERD may be an indicator of motor preparation, and could be used for objective evaluation of time estimation and motor timing. The minimal differences observed in the alpha changes in both paradigms suggest that alpha-ERD may not be linked to motor preparation. PMID- 12634489 TI - Ketamine alters neural processing of facial emotion recognition in healthy men: an fMRI study. AB - Disruption of facial emotion perception occurs in neuropsychiatric disorders where the expression of emotion is dulled or blunted, for example depersonalization disorder and schizophrenia. It has been suggested that, in the clinical context of emotional blunting, there is a shift in the relative contribution of brain regions subserving cognitive and emotional processing. The non-competitive glutamate receptor antagonist ketamine produces such emotional blunting in healthy subjects. Therefore, we hypothesised that in healthy subjects ketamine would elicit neural responses to emotional stimuli which mimicked those reported in depersonalization disorder and schizophrenia. Thus, we predicted that ketamine would produce reduced activity in limbic and visual brain regions involved in emotion processing, and increased activity in dorsal regions of the prefrontal cortex and cingulate gyrus, both associated with cognitive processing and, putatively, with emotion regulation. Measuring BOLD signal change in fMRI, we examined the neural correlates of ketamine-induced emotional blunting in eight young right-handed healthy men receiving an infusion of ketamine or saline placebo while viewing alternating 30 s blocks of faces displaying fear versus neutral expressions. The normal pattern of neural response occurred in limbic and visual cortex to fearful faces during the placebo infusion. Ketamine abolished this: significant BOLD signal change was demonstrated only in left visual cortex. However, with ketamine, neural responses were demonstrated to neutral expressions in visual cortex, cerebellum and left posterior cingulate gyrus. Emotional blunting may be associated with reduced limbic responses to emotional stimuli and a relative increase in the visual cortical response to neutral stimuli. PMID- 12634490 TI - Event-related potential evidence for attentional inhibition of return in audition. AB - Orienting attention to a spatial location facilitates responding to a subsequent target at that location, but inhibits the response if attention is oriented away from that location before the target appears there. This inhibitory effect of attention re-orienting, called inhibition of return (IOR), occurs in vision, hearing, touch, and cross-modally, and has been well studied behaviorally. However, little is known about its underlying neural mechanism(s). We report a study of the neural mechanism of auditory IOR using event-related potentials (ERPs). Auditory IOR was associated with elimination, but not reversal, of the Nd1 difference wave. Previous research indicates that the Nd1 represents an enhanced neural response to attended sounds. Also, auditory IOR was associated with a delay in the latency of the peak of the N1 component of the ERP at parietal sites. These effects are consistent with the accounts if inhibition of attention return that have been proposed for IOR, but are somewhat different from effects found in analogous ERP studies of visual IOR. PMID- 12634491 TI - Long-lasting facilitation by dehydroevodiamine. HClof synaptic responses evoked in the CA1 region of rat hippocampal slices. AB - We tested the effects of dehydroevodiamine.Cl (DHED) on field excitatory postsynaptic potentials (fEPSPs) evoked by the electrical stimulation of Schaffer collaterals-commissural fibres in the CA1 region of rat hippocampal slices. Bath applications of 10 microM DHED for 20 or 40 min induced long-lasting facilitation of fEPSPs, which outlasted the presence of DHED. A 10 min treatment with a higher concentration (100 microM) also induced long-lasting facilitation. The long lasting facilitation was blocked either by 10 microM atropine, the muscarinic receptor antagonist, or by 50 microM D-2-amino-5-phosphonopentanoic acid (D-AP5), an NMDA receptor antagonist. These results show that DHED produces long-lasting facilitation of synaptic transmission, and that this facilitation depends upon the activation of both the muscarinic and NMDA receptors. PMID- 12634492 TI - Inhibition of protein kinase A activity during conditioned taste aversion retrieval: interference with extinction or reconsolidation of a memory? AB - The involvement of the cAMP-dependent protein kinase A (PKA) signaling pathway in protein synthesis-dependent memory consolidation has been supported by studies of fear conditioning and conditioned taste aversion (CTA). The present experiment examined whether inhibition of PKA activity at the time of memory retrieval impedes or promotes subsequent extinction. When Rp-cAMPS was infused into the amygdala at the time of CTA testing (retrieval), extinction was accelerated. Results confirm recent findings that stored memories become more labile when they are retrieved and extend these findings to CTA memories. PMID- 12634493 TI - The eloquence of silent cortex: analysis of afferent input to deafferented cortex in arm amputees. AB - Cortical reorganisation after limb amputation includes topographic displacements of body representation areas and changes of areal extent. Remarkably, truncated nerves, which had innervated amputated limb parts and remained in the residual limbs, can retain access to the deafferented somatosensory cortex. Using somatosensory evoked potentials (SEP) we characterized afferences from electrically stimulated truncated nerves to the brachial plexus and cortex in 12 arm amputees. While peripheral responses were highly variable, thalamocortical input to S-1, as reflected by the primary cortical SEP component, was present in 11 of 12 patients. Despite long-term deafferentation, macroscopic phenomena of inhibition/refractoriness, as assessed by stimulus rate variations, appeared to be changed only marginally. Thus, deafferented cortex remains responsive when given artificial phantom input and could provide a neuronal substrate for spontaneous phantom limb sensations, including phantom pain. PMID- 12634494 TI - Interleukin-1 induces tau phosphorylation and morphological changes in cultured human astrocytes. AB - Tau phosphorylation is associated with neurite outgrowth and morphogenesis in neurons. Since inflammatory stimuli induce marked morphological changes in astrocytes, we examined the effect of interleukin 1alpha (IL-1alpha), an inflammatory cytokine, on the expression and phosphorylation of tau protein in cultured human astrocytes. Western blot analysis showed a rapid and transient increase (3-30 min) of tau phosphorylation at the AT8 epitope. The cells began to extend processes from 24 h after IL-1alpha stimulation. It is known that tau phosphorylation at the AT8 epitope reduces its ability to bind to, and stabilize, microtubules. The early tau phosphorylation induced by IL-1alpha might create a plastic environment for morphological changes in astrocytes. PMID- 12634495 TI - Stimulus frequency, calcium levels and striatal synaptic plasticity. AB - Electrophysiological and microfluorimetric measurements were combined to correlate the changes in intracellular calcium concentration to synaptic plasticity in striatal medium spiny projection neurons, during three different protocols of synaptic stimulation (1, 10, and 100 Hz). The 1 Hz stimulation protocol did not cause significant changes either in excitatory postsynaptic potential amplitude or in intracellular calcium concentration. The 10 Hz stimulation protocol induced a moderate increase of intracellular calcium without significantly affecting the excitatory postsynaptic potential amplitude. During the high frequency stimulation large, transient intracellular calcium elevations were recorded, and a significant long-term depression of excitatory postsynaptic potential was achieved. These results suggest that the induction of long-term depression required large, transient increases in intracellular calcium concentration. PMID- 12634496 TI - Expression of mRNA encoding the H1, H2, and H3 histamine receptors in the rat cochlea. AB - Histamine may have physiological functions in the inner ear. Histamine receptors, however, have not yet been identified in the mammalian cochlea. We determined, using reverse transcription polymerase chain reaction (RT-PCR), that H1, H2, and H3 histamine receptor mRNA is expressed in the rat cochlea. To investigate the distribution of each receptor, the cochlea was divided into three parts: the lateral portion, the medial portion, and the modiolus. The mRNA for each receptor subtype was detected in the modiolus but not in the lateral and medial portions of the cochlea. This is the first evidence of H1, H2, and H3 histamine receptor mRNA in the rat cochlea. These findings support the suggestion that histamine may play a physiological role in the cochlea. PMID- 12634497 TI - Age-related brain parenchymal fraction is significantly decreased in young multiple sclerosis patients: a quantitative MRI study. AB - The extent of brain atrophy was determined in 33 patients with multiple sclerosis (MS) and in 60 healthy subjects (21-76 years) by calculating brain parenchymal fractions (BPF, the ratio of brain parenchymal to intracranial volume) from 3D MRI. Within the normal data base, subjects at higher ages showed significantly lower BPF values. In younger MS patients, BPF was significantly decreased compared with age-matched controls (20-29 years, p= 0.0022; 30-39 years, =p 0.0001; 40-49 years,p = 0.0444) and was significantly correlated with disease duration and disease severity, but not with the number of detectable MS lesions. Determination of age-related BPF demonstrated significant brain atrophy in early MS and can be considered as a useful biological marker for monitoring MS. PMID- 12634498 TI - Amelioration of retarded neurite outgrowth of dorsal root ganglion neurons by overexpression of PKCdelta in diabetic rats. AB - To examine which isoform of protein kinase C (PKC) may be associated with impaired nerve regeneration in diabetes, we compared neurite outgrowth of isolated dorsal root ganglion (DRG) neurons in streptozocin (STZ)-induced diabetic and control rats. Neurite outgrowth was significantly retarded in diabetic neurons. Rottlerin, a PKCdelta specific inhibitor, significantly retracted neurite outgrowth whereas Go6976, an inhibitor specific for classical PKCs, had no effect, suggesting a significant role of PKCdelta in neurite outgrowth of DRG neurons. The expression of phosphorylated PKCdelta, but not total PKCdelta, in DRGs was decreased in diabetic rats. When this reduced expression was restored by overexpressing the PKCdelta in isolated DRG neurons, retardation of neurite outgrowth was significantly reversed in diabetic rats. These results suggest that a decrease in phosphorylated PKCdelta is at least in part responsible for impaired neurite outgrowth in diabetes, and that PKCdelta plays a significant role in the pathogenesis of diabetic neuropathy. This observation provides a useful clue for the treatment of diabetic neuropathy. PMID- 12634499 TI - The cortical substrate of visual extinction. AB - Neuroimaging studies investigated the attentional systems of the human brain revealing two networks, one for voluntary allocation of attention and another for stimulus-driven attentional processes. Whereas lesions of the latter system were supposed to lead to spatial neglect, we show that such lesions rather are typical for the occurrence of visual extinction. Extinction describes the inability of brain-damaged patients to detect a contralesional target in the presence of a competing ipsilesional stimulus. In a sample of consecutively admitted patients with right hemisphere stroke, we found dissociable cortical substrates for spatial neglect and visual extinction. There was a surprising congruency between the typical lesion site in patients with extinction and the activation clusters found in previous neuroimaging studies of healthy subjects. The results show that the temporo-parietal junction (TPJ), considered to be a crucial part of the stimulus-driven attentional network, is the neural substrate of visual extinction. PMID- 12634500 TI - Luciferase expression controlled by a viral gene promoter in a mammalian circadian pacemaker. AB - Retinal light exposure induces several immediate-early genes in the hypothalamic suprachiasmatic nucleus (SCN), which contains the major circadian pacemaker of mammals. Clock-controlled and light-induced genes expressed in the SCN such as c- and contain upstream regulatory elements similar to those of the major immediate early gene (IE-1) of the human cytomegalovirus. IE-1 expression is critical for viral reactivation from latency and increases in response to agents acting through depolarization or the cAMP response element. To test whether IE-1 could be under circadian control, bioluminescence was imaged in individual SCN cells of brain slice cultures from transgenic mice containing the IE-1 enhancer/promoter upstream from the firefly luciferase gene. A small percentage of the cells in neonatal and adult cultures displayed circadian transgene expression, particularly ones near the dorsomedial edge of the SCN. Single-cell bioluminescence imaging revealed that the circadian pacemaker can regulate exogenous viral genes and could play a role in viral diseases. PMID- 12634501 TI - Altered expression and distribution of FAC1 during NGF-induced neurite outgrowth of PC12 cells. AB - The molecular mechanisms by which neurotrophins such as nerve growth factor (NGF) induce neurite outgrowth and differentiation remain unclear, although multiple intracellular signaling pathways are known to participate. Recent studies have shown that nuclear transcription factors play an important role in NGF-stimulated neuritic outgrowth in PC12 cells. We investigated whether FAC1, a novel transcriptional regulator that exhibits altered subcellular distribution during brain development, is responsive to NGF-induced neurite outgrowth of PC12 cells. Our studies demonstrate that NGF induces a rapid, transient increase in FACI mRNA that is dependent upon ERK activation, and that FAC1 protein exhibits altered subcellular distribution during neurite outgrowth. These findings suggest that FAC1 expression and subcellular localization are regulated by NGF signaling pathways during neurite outgrowth. PMID- 12634502 TI - Coding of visual patterns and textures in monkey inferior temporal cortex. AB - Neural coding for texture features of visual objects was investigated in monkey inferior temporal cortex by inactivating intrinsic GABAergic inhibition. The inactivation enabled a substantial number of cells to respond to originally ineffective texture pattern that had a particular feature distinct from the originally effective pattern, or to ineffective texture and non-texture stimuli that possessed a component feature of the originally effective texture. Cells that showed selectivity changes related to a texture feature were often met along a vertical recording track. We suggest that a texture feature is coded by a group of cells which signal different aspects of the texture. The coding occurs at different processing levels, from extracting a particular feature within patterns to detecting complex texture features combined with color and shape of natural objects. PMID- 12634503 TI - Subdivisions of the dorsal raphe nucleus projecting to the lateral geniculate nucleus and primary visual cortex in the Mongolian gerbil. AB - The mammalian dorsal raphe nucleus (DRN) is composed of sub-divisions with different anatomical and functional properties. Using cholera toxin subunit B as a retrograde tracer, DRN subdivisions projecting to the lateral geniculate nucleus and to the primary visual cortex were examined in the Mongolian gerbil. DRN neurons projecting to the lateral geniculate nucleus were observed in the lateral DRN (rostrally) and in the ventromedial DRN (caudally), while DRN cells projecting to the primary visual cortex were observed at all rostral-caudal levels in the ventromedial DRN. These results demonstrate a significant overlap between the DRN projections to the lateral geniculate and superior colliculus, and show that only the caudal ventromedial DRN projects to all three major visual targets: the lateral geniculate nucleus, primary visual cortex, and superior colliculus. Since the DRN is involved in depression and other neuropsychiatric disorders, as well as is affected by many psychotropic substances, these data may help to develop new treatments and therapies targeting specific DRN subdivisions. PMID- 12634504 TI - Dose-response of cyclosporin A in attenuating traumatic axonal injury in rat. AB - Cyclosporin A has emerged as a promising therapeutic agent in traumatic brain injury (TBI), although its precise neuroprotective mechanism is unclear. Cyclosporin A, given as a single-dose intrathecal bolus, has previously been shown to attenuate mitochondrial damage and reduce axonal injury in experimental TBI. We assessed the effect of a range of intravenous cyclosporin A doses upon axonal injury attenuation to determine the ideal dose. Rats were subjected to experimental TBI and given one of five intravenous doses of cyclosporin A. At 3 h post-injury, brains were processed for brain tissue cyclosporin A concentration. In a second set of animals, at 24 h postinjury, brains were processed for amyloid precursor protein immunoreactivity, a widely used marker of axonal injury. Intravenous administration produced therapeutic levels of cyclosporin A in brain parenchyma. Higher concentrations were achieved with equivalent doses given intrathecally; this is consistent with the reported poor blood-brain barrier permeability of cyclosporin A. Cyclosporin A 10 mg/kg i.v. produced the greatest degree of neuroprotection against diffuse axonal injury; cyclosporin A 50 mg/kg i.v. was toxic. Intravenous cyclosporin A administration achieves therapeutic levels in brain parenchyma and 10 mg/kg is the most effective dose in attenuating axonal damage after traumatic brain injury. PMID- 12634505 TI - Haptic texture affects the kinematics of pointing movements, but not of eye movements. AB - Discrepant findings on the degree of eye-hand coupling suggest its dependence on the task. One task characteristic modulating this coupling may be the relevance of certain target attributes for each motor system. We tested this assumption by comparing eye and hand movements towards targets of different haptic texture, a target attribute which is behaviourally relevant only to the hand, not the eye. Pointing to a slippery target (fur) resulted in longer hand movement time than to a rougher target (sandpaper). This effect was due to an increased ratio of time spent in deceleration. In contrast, eye movement time was invariant across different haptic target textures. Thus, information about target texture is used differently by eye and hand. PMID- 12634506 TI - Levetiracetam reduces caffeine-induced Ca2+ transients and epileptiform potentials in hippocampal neurons. AB - The effect of the novel antiepileptic drug levetiracetam on caffeine (10 mM) induced intracellular calcium ([Ca2+]i) response was investigated in rat hippocampal neurons in culture, with the aim of exploring the cellular mechanisms of this new drug. Levetiracetam significantly reduced caffeine-induced [Ca2+]i) response, with maximum inhibition at 32 microM. The R-enantiomer of levetiracetam, ucb L060, which is devoid of anticonvulsant activity, at 32 microM had no effect on caffeine-induced [Ca2+]i) response. Caffeine 10 mM also induced epileptiform field potentials in rat hippocampal slices : single stimuli evoked repetitive population spikes and spontaneous field bursts regularly occurred. Levetiracetam (32 microM) significantly inhibited the amplitudes and the number of caffeine-induced repeated population spikes and delayed the appearance of spontaneous bursts, while ucb L060 (32 microM) completely lacked anti-caffeine activity. These results suggest that the inhibition of caffeine-induced Ca release from intra-neuronal stores might be an excitability-reducing effect of levetiracetam, contributing to its antiepileptic activity. PMID- 12634507 TI - Gamma-band EEGs predict autonomic responses during mental arithmetic. AB - Relationship between gamma-band (20-80 Hz) EEGs and autonomic functions (heart rate, blood pressure, instantaneous lung flow) in a mental arithmetic task (MAT) were investigated. Subjects were instructed to add 2 integers after presentation of audio voices indicating the integers in the MAT, and to listen to the same voices without mental arithmetic in the control task. The gamma-EEGs and autonomic functions coherently changed in response to the voices before mental arithmetic in the MAT. Furthermore, an increase in gamma-EEGs went ahead of the autonomic fluctuation. The results suggest that the gamma-band activity modulates and adjusts peripheral autonomic functions in advance before actual mental activity. This is the first demonstration that cortical gamma-band activity triggers peripheral autonomic responses during mental activity. PMID- 12634508 TI - Injections of the neuroprotective peptide NAP to newborn mice attenuate head injury-related dysfunction in adults. AB - The prophylactic neuroprotective effects of NAP, a femtomolar-acting neuroprotective peptide were tested in a mouse model of head trauma. NAP was injected for the first 3 weeks of life and head injury was initiated at 4 months. After trauma, mice were tested for their performance by evaluating damaged motor ability, balance and alertness. Comparison of the performance 1 h and 1 week after injury indicated that NAP treatment resulted in faster and enhanced recovery. In a 5-day Morris water maze test with mice suffering moderate to severe injuries, only the NAP-treated group learned to find the hidden platform in the maze. Furthermore, NAP treatment resulted in decreased mRNA expression of the inflammation marker, Mac-1. Thus, a potentially new prophylactic treatment against neurodegeneration is suggested. PMID- 12634509 TI - Investigation of mitochondrial function in hereditary spastic paraparesis. AB - Following the association of hereditary spastic paraparesis (HSP) with mutation in the paraplegin gene (SPG7) and mitochondrial dysfunction, we wished to investigate whether mitochondrial dysfunction might be associated with other forms of HSP. Five cases of HSP caused by mutation in the spastin gene (SPG4) and nine cases with HSP with mutation in the spastin and paraplegin genes excluded (non-SPG4/SPG7), were investigated for mitochondrial dysfunction. Muscle tissue from the HSP groups and a control group was analysed histochemically and spectrophotometrically for mitochondrial dysfunction. A significant decrease in mitochondrial respiratory chain complexes I and IV was demonstrated in the non SPG4/SPG7 group. No abnormality was detected in the SPG4 group. We therefore conclude that there is evidence for mitochondrial dysfunction in non-SPG4/SPG7 HSP. There is no evidence for mitochondrial dysfunction in the pathogenesis of spastin-related HSP. PMID- 12634510 TI - Covert and overt orienting to gaze direction cues and the effects of fixation offset. AB - We examined covert and overt orienting in response to non-predictive gaze direction cues and investigated whether the subcortical superior colliculus (SC) plays a role in this type of orienting. Participants viewed a centrally presented gazing schematic face and responded to targets appearing at gazed-at or non-gazed at locations either by making a keypress response while maintaining central fixation or by making an eye movement to the target. For both response conditions, the fixation stimulus (the gazing face) either remained on the screen or was extinguished at the time of target presentation, a manipulation known to engage and disengage the SC. Results revealed that participants making manual responses oriented covertly to the gazed-at location regardless of the fixation condition, and that participants making eye movements oriented overtly only if the fixation stimulus remained on the screen. Overt gaze-triggered orienting was not enhanced relative to covert orienting, and the fixation offset effect was not reduced for averted gaze cues relative to straight gaze cues. These findings suggest that gaze direction cues do not activate or predisengage the oculomotor system, and thus that orienting to gaze direction does not engage the SC. This is consistent with the view that gaze-triggered orienting is a unique form of reflexive orienting that depends crucially on cortical processes. PMID- 12634511 TI - Training with cognitive sequences improves syntactic comprehension in agrammatic aphasics. AB - A major open question in cognitive neuroscience concerns the modularity of language: does human language rely, in part, on neural processes that are not language specific? Such reliance would predict that learning should transfer between non-linguistic and linguistic domains via this common neural basis. To test this prediction, we studied effects of non-linguistic cognitive sequence training on syntactic comprehension of six left-hemisphere damaged aphasic patients. Syntactic comprehension impairment was quantified before and after 10 weeks of training on a non-linguistic sequence processing task. This task used a transformational rule specifically corresponding to the transformation required for understanding a particular type of sentence referred to as relativised. Non linguistic sequencing improved significantly with training (day effect: F(9,45)=3.7, <0.005). Moreover, a significant transfer of this improvement was observed for relativised, but not active nor passive sentences (pre-post x type interaction: F(2,10)=4.72, <0.05). The specificity of this transfer indicates that language relies partially on functional and neural processes that are not language specific. PMID- 12634512 TI - Altered hemispheric asymmetry of auditory N100m in adults with developmental dyslexia. AB - The current study aimed at determining whether the deviance of hemispheric asymmetry in the auditory cortex of children with dyslexia is also evident in dyslexic adults. Ten adult dyslexic subjects and 10 normally literate controls were presented with the syllable [ba:] while event-related brain activity was recorded from both hemispheres using whole-head magnetoencephalography. In control subjects, the auditory N100m source was found to be asymmetrical with a more anterior localization in the right than in the left perisylvian cortex (difference = 0.78 cm). The N100 m dipoles in dyslexic adults did not exhibit the same interhemispheric asymmetry (difference = -0.06 cm). The results indicate that reduced hemispheric laterality of perisylvian regions in dyslexia persists into adulthood. PMID- 12634513 TI - Do atypical antipsychotics fail to exert cognitive sparing effects? AB - The aim of the study was to investigate patterns of the P600 component of event related potentials (ERPs) elicited during a working memory test in 16 male schizophrenic patients experiencing auditory hallucinations before and after treatment with clozapine and olanzapine, and 13 male normal subjects matched for age and educational level. Before treatment, patients showed significantly reduced P600 amplitudes on the right parietal region compared with controls, and when in remission also showed significantly reduced P600 amplitudes located on the right parietal and temporofrontal areas, compared both to themselves before treatment and to normal controls. The patient's memory performance before and after treatment remained significantly lower than that of healthy controls. These findings may indicate that auditory hallucinations in schizophrenia are associated with impaired synchronization of the processes related to target detection, as reflected by the P600. The present study also casts doubts regarding the cognitive sparing effect of atypical antipsychotics, despite the fact that they mediate symptom improvement. PMID- 12634514 TI - Language recovery following subthalamic nucleus stimulation in Parkinson's disease. AB - Neuropsychological consequences of stimulation of the subthalamic nucleus for treatment of drug-resistant Parkinson's disease (PD) have been studied previously. However, no detailed investigations of linguistic function modifications have been carried out. We studied four consecutive patients with PD who underwent chronic bilateral stimulation of the subthalamic nuclei. Neuropsychological and linguistic evaluations were performed before and 2 weeks after surgery. Linguistic abilities were studied also 1 year after surgery with stimulators both off and on. Intraphrasal hesitation pauses, phonemic paraphasias and morpho-syntactic errors were significantly reduced and lexical retrieval improved with stimulation of the subthalamic nuclei. Implicit linguistic phenomena, mainly occurring within basal ganglia circuitry, benefited by recovery of functional equilibrium within basal nuclei and between overall basal ganglia circuitry and cerebral cortex. PMID- 12634515 TI - Sipatrigine and oligodendrocyte and axonal pathology following transient focal cerebral ischaemia in the rat. AB - The ability of Sipatrigine (BW619c89), a sodium and calcium channel blocker to protect neuronal perikarya, axons and oligodendrocytes from ischaemia has been evaluated in the rat. Sipatrigine (30 mg/kg) was administered 15 min prior to transient (2 h) occlusion of the middle cerebral artery with animals being sacrificed 24 h after induction of ischaemia. Sipatrigine significantly reduced the volumes of neuronal perikarya damage (by 35%) and oligodendrocyte damage (by 54%). Sipatrigine minimally altered axonal damage (median scores: vehicle approximately 30, Sipatrigine approximately 26). The present report provides evidence that neuronal perikarya, axons and oligodendrocytes may be differentially protected by pharmacological intervention in focal cerebral ischaemia. PMID- 12634516 TI - Context-dependent song amplitude control in Bengalese finches. AB - Estrildid finches have two song types: directed (courtship) and undirected (solo). These are acoustically identical and differ only in social context. Recent studies have shown that undirected singing is accompanied by strong activation of the basal ganglia pathway, whereas directed singing is not, which suggests a different degree of feedback control between the two. We examined whether the magnitude of the Lombard effect, i.e., vocal amplitude regulation in response to environmental noise, differed based on the song context. Our results indicate that Bengalese finches change the amplitude of undirected song based on background noise levels, but do not do so for directed song. This is the first behavioral evidence suggesting that feedback control of song output vary by social context. PMID- 12634517 TI - Differential involvement of episodic and face representations in ERP repetition effects. AB - The purpose of this study was to disentangle the contribution of episodic perceptual from pre-existing memory representations of faces to repetition effects. ERPs were recorded to first and second presentations of same and different photos of famous and unfamiliar faces, in an incidental task where occasional non-targets had to be detected. Repetition of same and different photos of famous faces resulted in an N400 amplitude decrement. No such N400 repetition-induced attenuation was observed for unfamiliar faces. In addition, repetition of same photos of faces, and not different ones, gave rise to an early ERP repetition effect (starting at approximately 350 ms) with an occipito temporal scalp distribution. Together, these results suggest that repetition effects depend on two temporally and may be neuro-functionally distinct loci, episode-based representation and face recognition units stored in long-term memory. PMID- 12634518 TI - G protein beta3 subunit 825T allele is associated with depression in young, healthy subjects. AB - This study investigated the relationship between the G protein beta3 subunit 825T allele and depression in young, healthy subjects. 825T and 825C allele carriers were characterized among 190 healthy medical students. State depression was assessed by the Becks Depression Inventory, supplemented by subscales of the Freiburg Personality Inventory, and Questionnaire for Measuring Factors of Aggression which assess trait depression. Depression of homo- and heterozygous 825T allele carriers was compared to 825C allele carriers via multi-level statistical analysis. 825T allele carriers displayed higher levels of depression, as measured by each questionnaire. Regression analysis demonstrated that allele type was the single predictor of depression. The predictive capacity of the 825T allele in depression was further supported by odds-ratio analysis. We conclude that the G protein beta3 subunit 825T allele is predictive of depressive mood in a young, healthy population. PMID- 12634519 TI - Renal injury and operative management in the United States: results of a population-based study. AB - BACKGROUND: To evaluate the extent to which nonoperative renal trauma management has been adopted, we determined the incidence of renal injury and the rate of operative management across the United States. METHODS: International Classification of Diseases, Ninth Revision diagnosis and procedure codes identified patients with renal injuries in an 18-state administrative database representing 62% of the U.S. population. RESULTS: Of 523,870 patients hospitalized for trauma in 1997 or 1998, 6,231 (1.2%) had renal injuries (4.89 per 100,000 population). Sixty-four percent of patients with injuries that were classified had contusions/hematomas, 26.3% had lacerations, 5.3% had parenchymal disruption, and 4% had vascular injuries. Eleven percent of renal trauma patients required surgical management of their kidney injuries, of whom 61%, or 7% of patients with renal injuries overall, underwent nephrectomy. Injury Severity Score, mechanism, and renal injury severity were independent predictors of nephrectomy. CONCLUSION: The nephrectomy rate in community and academic centers reflects renal and global injury severity. Prospective trials are indicated to determine whether, in the traumatized patient with severe kidney injury, renal preservation could lead to improved outcomes compared with nephrectomy. PMID- 12634520 TI - Traumatic rupture of the urinary bladder: is the suprapubic tube necessary? AB - BACKGROUND: Although surgical principles are well accepted for the treatment of an intraperitoneal or extraperitoneal rupture of the urinary bladder, the type and number of drainage catheters needed to obtain a satisfactory outcome with minimal patient morbidity have yet to be determined. METHODS: This was a retrospective review of data on injured patients with the diagnosis of an intraperitoneal or extraperitoneal rupture of the urinary bladder from penetrating or blunt trauma. RESULTS: Of the 51 patients identified, 28 were treated with suprapubic and transurethral catheters, whereas 23 received a transurethral catheter only. Complications and catheter duration times were similar regardless of type of bladder injury or drainage catheter used (p > 0.5). CONCLUSION: These data suggest that there are similar outcomes and complication rates for patients treated with suprapubic and transurethral catheters versus transurethral catheter only. Transurethral catheters alone seem effective in draining all types of bladder injuries. PMID- 12634521 TI - External fixation or arteriogram in bleeding pelvic fracture: initial therapy guided by markers of arterial hemorrhage. AB - BACKGROUND: Bleeding pelvic fractures (BPF) carry mortality as high as 60%, yet controversy remains over optimal initial management. Some base initial intervention on fracture pattern, with immediate external fixation (EX FIX) in amenable fractures aimed at controlling venous bleeding. Others feel ongoing hemodynamic instability indicates arterial bleeding, and prefer early angiography (ANGIO) before EX-FIX. Our aim was to evaluate markers of arterial bleeding in patients with BPF, thus identifying patients requiring early ANGIO regardless of fracture pattern. METHODS: Patients with pelvis fracture were identified from a Level I trauma center registry over a 7-year period and records reviewed. From this group, two subsets were analyzed: those with initial hypotension related to pelvic fracture, and those without hypotension who underwent pelvic ANGIO. Data included hemodynamics, response to resuscitation, presence of contrast blush on CT, fracture treatment and outcome. Adequate response to initial resuscitation (R) was defined as a sustained (>2 hours) improvement of systolic blood pressure to >90 mm Hg systolic after the administration of < or = 2 units packed red blood cells. Those with repeated episodes of hypotension despite resuscitation were classified as non-responders (NR) RESULTS: From 1/94-1/01, 1171 patients were admitted with pelvic ring fracture. Thirty-five (0.3%) had hypotension attributable to pelvis fracture. 28 fell into the NR group, and 26 of these underwent ANGIO. Nineteen (73%) showed arterial bleeding while 3 resuscitation response patients underwent ANGIO with none demonstrating bleeding (p = 0.03). Sensitivity and specificity of inadequate response to initial resuscitation for predicting the presence of arterial bleeding on ANGIO were 100% and 30% respectively while negative and positive predictive value were 100% and 73%. In patients with fractures amenable to external fixation (n = 16), 44% had arterial bleeding on ANGIO, and all were in the NR group. An additional 17 patients without hypotension also underwent ANGIO. Contrast blush on admission CT was seen in 4, 3 of which had arterial bleeding seen on ANGIO (75%). Sensitivity and specificity for contrast blush in predicting bleeding on ANGIO were 60% and 92% with positive and negative predictive value being 75% and 85%. CONCLUSIONS: In patients with hypotension and pelvic fracture, therapy selection based on initial response to resuscitation in BPF yields a 73% positive ANGIO rate in NR patients. Delay in ANGIO for EX FIX in patients with amenable fractures would have delayed embolization in the face of ongoing arterial bleeding in 44% of patients. In stable patients with pelvic fracture, contrast blush also indicates a high likelihood of arterial injury and ANGIO is indicated. Optimal therapy in the face of BPF requires early determination of the presence of arterial bleeding so that ANGIO can be rapidly obtained, and response to initial resuscitation as well as the presence of contrast blush aid in this decision. PMID- 12634522 TI - The effect of paramedic rapid sequence intubation on outcome in patients with severe traumatic brain injury. AB - OBJECTIVE: To evaluate the effect of paramedic rapid sequence intubation (RSI) on outcome in patients with severe traumatic brain injury. METHODS: Adult major trauma victims were prospectively enrolled over two years using the following inclusion criteria: Glasgow Coma Scale (GCS) 3-8, suspected head injury by mechanism or physical examination, transport time > 10," and inability to intubate without RSI. Midazolam and succinylcholine were administered before laryngoscopy; rocuronium was given after tube placement was confirmed using physical examination, capnometry, syringe aspiration, and pulse oximetry. The Combitube was used as a salvage airway device. For this analysis, trial patients were excluded for absence of a head injury (Head/Neck AIS score < 2), failure to fulfill major trauma outcome study criteria, unsuccessful intubation or Combitube insertion, or death in the field or in the resuscitation suite within 30" of arrival. Each study patient was hand matched to three nonintubated historical controls from our trauma registry using the following parameters: age, sex, mechanism of injury, trauma center, and AIS score for each body system. Controls were excluded for Head/Neck AIS defined by a c-spine injury or death in the field or in the resuscitation suite within 30" of arrival. chi 2, odds ratios, and logistic regression were used to investigate the impact of RSI on the primary outcome measures of mortality and incidence of a "good outcome," defined as discharge to home, rehabilitation, psychiatric facility, jail, or signing out against medical advice. RESULTS: A total of 209 trial patients were hand matched to 627 controls. The groups were similar with regard to all matching parameters, admission vital signs, frequency of specific head injury diagnoses, and incidence of invasive procedures. Mortality was significantly increased in the trial cohort versus controls for all patients (33.0% versus 24.2%, p < 0.05) and in those with Head/Neck AIS scores of 3 or greater (41.1% versus 30.3%, p < 0.05). The incidence of a "good outcome" was lower in the trial cohort versus controls (45.5% versus 57.9%, p < 0.01). Factors that may have contributed to the increase in mortality include transient hypoxia, inadvertent hyperventilation, and longer scene times associated with the RSI procedure. CONCLUSION: Paramedic RSI protocols to facilitate intubation of head-injured patients were associated with an increase in mortality and decrease in good outcomes versus matched historical controls. PMID- 12634523 TI - Minimizing dilutional coagulopathy in exsanguinating hemorrhage: a computer simulation. AB - BACKGROUND: Current massive transfusion guidelines are derived from washout equations that may not apply to bleeding trauma patients. Our aim was to analyze these guidelines using a computer simulation. METHODS: A combined hemodilution and hemodynamic model of an exsanguinating patient was developed to calculate the changes in prothrombin time (PT), fibrinogen, and platelets with bleeding. The model was calibrated to data from 44 patients. Time intervals to subhemostatic values of each coagulation test were calculated for a range of replacement options. RESULTS: Prolongation of PT is the sentinel event of dilutional coagulopathy and occurs early in the operation. The key to preventing coagulopathy is plasma infusion before PT becomes subhemostatic. The optimal replacement ratios were 2:3 for plasma and 8:10 for platelets. Concurrent transfusion of plasma with blood is another effective strategy for minimizing coagulopathy. CONCLUSION: Existing protocols underestimate the dilution of clotting factors in severely bleeding patients. The model presents an innovative approach to optimizing component replacement in exsanguinating hemorrhage. PMID- 12634524 TI - The association between gender and mortality among trauma patients as modified by age. AB - BACKGROUND: Several studies have reported a null association between gender and mortality after traumatic injury, whereas others found an age-specific association between male gender and increased mortality. Relatively small sample sizes may have contributed to the heterogeneity among existing studies; therefore, a large-sample-size study was undertaken. METHODS: The National Trauma Data Bank was queried, yielding data for over 150,000 patients involved in blunt or penetrating trauma. Crude and adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were calculated for the association between gender and mortality, both overall and according to mechanism of injury and age categories. RESULTS: Among those who sustained blunt trauma, male patients had a significant increase in the risk of death compared with female patients (OR, 1.49; 95% CI, 1.39-1.59) that was most apparent for those > or = 50 years of age (OR, 1.97; 95% CI, 1.84 2.11). For penetrating trauma patients, essentially no significant association, either overall (OR, 1.03; 95% CI, 0.91-1.17) or by age category, was apparent. CONCLUSION: This study found an association between gender and mortality among blunt trauma patients, particularly those aged > or = 50 years. Animal studies demonstrate that the sex hormones influence the inflammatory response to injury. These results may highlight the importance of sex hormones in traumatic injury outcomes. PMID- 12634525 TI - Diagnosis of posttraumatic pulmonary embolism: is chest computed tomographic angiography acceptable? AB - BACKGROUND: Pulmonary angiography (PA-gram) has long been the accepted criterion standard for diagnosing pulmonary embolism (PE). Computed tomographic angiography has recently been advocated as an equivalent alternative to PA-gram. CT angiography is known to be insensitive for peripheral (segmental and subsegmental) emboli. We have previously found that a significant number of posttraumatic PEs occur early. We therefore hypothesized that because of the fragmentation of these early (soft) clots, posttraumatic PEs would be found disproportionately in the lung periphery. METHODS: Trauma patients with PE confirmed by PA-gram were identified from our trauma database and medical records. PA-grams and reports were re-reviewed and the location of all emboli was documented. RESULTS: We identified 45 patients, with an average age of 46 +/- 19 years; two thirds of the patients were men and 82% had a blunt mechanism of injury. Patients had PE diagnosed between days 0 and 57. Overall, PE was confined to segmental or smaller vessels in 27 (60%) patients and to subsegmental vessels in 7 (16%) patients. Twelve patients (27%) had a PE within the first 4 days. Furthermore, 32 patients (71%) had unilateral clot and 22 patients (48.9%) had clot confined to one region. CONCLUSION: PE frequently occurs soon after injury. The majority of PEs after trauma are found peripherally (in segmental or subsegmental vessels). Right/left pulmonary artery embolisms are likely to be found only later in a trauma patient's course. Any diagnostic study used to diagnose pulmonary embolism in trauma patients must have sufficient resolution capacity to reliably detect segmental and subsegmental clot. A diagnostic modality such as CT scanning that is insensitive to peripheral embolisms may miss a significant number of posttraumatic PEs. PMID- 12634526 TI - Elderly trauma patients with rib fractures are at greater risk of death and pneumonia. AB - BACKGROUND: The purpose of this study was to show that elderly patients admitted with rib fractures after blunt trauma have increased mortality. METHODS: Demographic, injury severity, and outcome data on a cohort of consecutive adult trauma admissions with rib fractures to a tertiary care trauma center from April 1, 1993, to March 31, 2000, were extracted from our trauma registry. RESULTS: Among 4,325 blunt trauma admissions, there were 405 (9.4%) patients with rib fractures; 113 were aged > or = 65. Injuries were severe, with Injury Severity Score (ISS) > or = 16 in 54.8% of cases, a mean hospital stay of 26.8 +/- 43.7 days, and 28.6% of patients requiring mechanical ventilation. Mortality (19.5% vs. 9.3%; p < 0.05), presence of comorbidity (61.1% vs. 8.6%; p < 0.0001), and falls (14.6% vs. 0.7%; p < 0.0001) were significantly higher in patients aged > or = 65 despite significantly lower ISS (p = 0.031), higher Glasgow Coma Scale score (p = 0.0003), and higher Revised Trauma Score (p < 0.0001). After adjusting for severity (i.e., ISS and Revised Trauma Score), comorbidity, and multiple rib fractures, patients aged > or = 65 had five times the odds of dying when compared with those < 65 years old. CONCLUSION: Despite lower indices of injury severity, even after taking account of comorbidities, mortality was significantly increased in elderly patients admitted to a trauma center with rib fractures. PMID- 12634527 TI - Long-term outcomes after injury in the elderly. AB - BACKGROUND: The lasting impact of injury on lifestyle in the elderly remains poorly defined. The purpose of this study was to determine the long-term quality of-life outcomes in elderly trauma patients. METHODS: The trauma registry at a regional trauma center was used to identify hospital survivors of injury > or = 65 years old discharged from April 1996 to March 1999. The 36-Item Short Form (SF 36) Health Survey was administered to this group by telephone interview and the scores compared with age-adjusted Canadian norms. Comparisons with test were made for continuous data. RESULTS: Complete data collection was achieved in 128 of 171 (75%) study patients. The mean Injury Severity Score was 21, the mean initial Glasgow Coma Scale score was 13, and the mean age was 74. Most (97%) were victims of blunt trauma. Compared with Canadian age-adjusted norms, there was a significant (p < 0.05) decrease in seven of eight SF-36 domains: Physical Functioning, Role-Physical and Role-Emotional (limitations secondary to physical and emotional health), Social Functioning, Mental Health, Vitality, and General Health. Before injury, most (98%) were living independently at home. However, at long-term follow-up (mean, 2.8 years; range, 1.5-4.5 years), only 63% were living independently and 20% still required home care. CONCLUSION: Although the majority of elderly injury survivors achieve independent living, long-term follow-up indicates significant residual disability in quality of life as measured by the SF-36. PMID- 12634528 TI - Time to deterioration of the elderly, anticoagulated, minor head injury patient who presents without evidence of neurologic abnormality. AB - BACKGROUND: Generally accepted guidelines regarding the care of the elderly, anticoagulated minor head injury patient do not exist within the trauma literature. METHODS: Charts were reviewed on all anticoagulated, minor head injury patients older than 65 years between January 1993 and May 2000. Postinjury course was examined for neurologic changes, times, coagulation/radiographic studies, reversal, operative intervention, and outcome. RESULTS: Thirty-two patients were identified. Twenty-four patients were discharged from the Emergency Department. Three of the remaining eight patients had initial Glasgow Coma Scale scores of 15, 15, and 14 but became comatose over a mean course of 3.83 hours. A fourth patient presented comatose 6 hours postinjury, down from "acting normal." Three of these four patients died. CONCLUSION: Elderly, anticoagulated patients with minor head trauma risk neurologic deterioration within 6 hours of injury, despite an initially normal neurologic examination. Early cranial computed tomographic scanning and close observation for a minimum of 6 hours are indicated. PMID- 12634529 TI - Patients with a head injury who "talk and die" in the 1990s. AB - BACKGROUND: Patients who "talk and die" after head injury may represent a group who suffer delayed and therefore potentially preventable complications after injury. We have compared the clinical and pathologic features of patients who talk and die with those who "talk and live" after head injury. METHODS: Data collected prospectively by the Scottish Trauma Audit Group were used to identify patients with a head injury and classify them according to verbal response at admission to hospital. All "talking" patients in the catchment area of a regional neurosurgical center were selected and those who died were compared with those who survived. RESULTS: Seven hundred eighty-nine talking patients were identified. Seven hundred twenty-seven patients survived and 62 died. Patients who talked and died were older, had more severe extracranial injuries, had lower consciousness levels, and reached theater more quickly than those who talked and lived. Thirty-one of the patients that died had extra-axial hematomas. CONCLUSION: Even with increased availability of computed tomographic scanning, some patients still talk and die after head injury. PMID- 12634530 TI - Predicting the need for laparotomy in pediatric trauma patients on the basis of the ultrasound score. AB - BACKGROUND: It is possible to quantify the amount of hemoperitoneum seen on focused assessment with sonography for trauma (FAST) using a simple scoring system that had previously been shown to correlate with the need for subsequent laparotomy in adults. A score of 3 or greater was shown to be highly accurate in predicting the need for laparotomy. We hypothesized that this scoring system might also predict the need for laparotomy in pediatric trauma patients. METHODS: We retrospectively reviewed all records for patients 15 years and younger who underwent FAST after blunt trauma. A "positive" ultrasound examination was defined as one containing free intraperitoneal fluid with or without solid organ injury. The ultrasound score (USS) was defined as the depth of the deepest pocket of fluid collection measured in centimeters plus the number of additional spaces where fluid was seen. RESULTS: Thirty-eight (19.6%) of 193 patients who had FAST performed had positive ultrasound examinations. Thirty-seven patients with complete records were analyzed. There were no differences between patients with a USS < or = 3.0 and those with a USS > 3.0 in terms of admission pulse, Glasgow Coma Scale score, Injury Severity Score, or the proportion of patients who were initially hypotensive. One of 22 patients with a USS < or = 3.0 required therapeutic laparotomy versus 8 of 15 patients with a USS > 3.0 ( = 0.002). For a USS > 3.0, sensitivity, specificity, and accuracy in predicting therapeutic laparotomy were 89%, 75%, and 78%, respectively. CONCLUSION: Ultrasound quantification of hemoperitoneum by a simple scoring system may serve as a useful adjunct to traditional clinical parameters in predicting the need for subsequent laparotomy in pediatric patients. Prospective validation with a larger study is required. PMID- 12634531 TI - Correlation of clinical characteristics and outcomes with injury scoring in blunt cardiac trauma. AB - BACKGROUND: Clinical sequelae from blunt cardiac trauma (BCT) may range from minor electrocardiographic abnormalities to death from free-wall rupture. There are no established clinical characteristics or injury scoring systems that are able to predict survival in these patients. METHODS: A retrospective review of medical records from a Level I trauma center identified 47 patients with BCT. A grade assigned on the basis of the American Association for the Surgery of Trauma Organ Injury Scale (OIS) was assigned to each case studied. Clinical data, including the Injury Severity Score (ISS), and outcomes were analyzed for association with OIS grade. RESULTS: The average ISS was 27.9, and the overall mortality rate was 31.9%. The majority of patients were either grade II or IV, with the latter having the highest mortality. Hypotension at admission, cardiac arrest, lack of vital signs at admission, ISS, hours to diagnosis, and death all had significant association with assigned OIS grade. Factors associated with mortality included ISS; OIS grade; shorter time to diagnosis; cardiac tamponade; cardiac rupture; lack of vital signs at admission; and concomitant injury to either the thoracic aorta or to the liver, spleen, or kidneys. CONCLUSION: The OIS grade, assigned on the basis of anatomic site of injury and electrocardiographic abnormalities, appears to correlate with severity of injury and survival. Although injury scoring should not be used exclusively to guide management in trauma patients, the grading system studied may be useful in predicting outcomes in patients with BCT. PMID- 12634532 TI - Surgical treatment of avulsion fracture of the posterior cruciate ligament and postoperative management. AB - BACKGROUND: Avulsion fractures of the posterior cruciate ligament have long been regarded as rare injuries. In the past, it was common practice to use cast immobilization as an external adjunct after open reduction and internal fixation of fractures. METHODS: Sixteen patients with displaced avulsion fractures of the posterior cruciate ligament were treated with open reduction and internal fixation between August 1989 and July 1993. Malleolar screws were chosen as fixation devices in 14 patients. In the other two, pull-through sutures were used because the size of the fractured fragments was too small to obtain purchase of screws. The postoperative management protocol evolved from an initial regimen of 6 weeks' immobilization in a cast with the knee flexed to 40 degrees for the first five patients (group I), to 4 weeks' immobilization in a cast for the next six patients (group II), to the present protocol of immediate postoperative range of motion (40-70 degrees) with muscle-strengthening exercises in a functional brace for the last five patients (group III). The average follow-up period was 36 months (range, 24-58 months). Hughston's criteria were used to assess the clinical results. RESULTS: Overall, there were 12 (75%) good and 4 fair (25%) results. There was no poor result. CONCLUSION: Avulsion fractures of the posterior cruciate ligament should be treated with open reduction and stable internal fixation if any displacement is seen on initial radiographs at presentation. With the use of functional brace and aggressive postoperative rehabilitation program (i.e., immediate range of motion of 40-70 degrees with muscle-strengthening exercises), satisfactory results can be expected and achieved. PMID- 12634533 TI - Traumatic dislocation and fracture-dislocation of the hip: a long-term follow-up study. AB - OBJECTIVE: Traumatic dislocation and fracture-dislocation of the hip is an absolute orthopedic emergency that is steadily increasing in incidence. Early recognition and prompt, stable reduction is the essence of successful management. A delay in recognition and reduction leads to preventable complications and morbidity. The purpose of this retrospective study is to identify prognostic factors that predict long-term outcome after hip dislocation. METHODS: Between 1980 and 1994, 107 patients with traumatic dislocation of the hips were treated, and 62 are reviewed in this study. There were 57 posterior fracture-dislocations and 5 anterior-obturator dislocations. All of the patients' charts were reviewed. The physical examinations and radiologic controls of the patients who were called for last follow-up examination were performed by the first two authors (V.S. and E.K.). Anterior and posterior fracture-dislocations were classified according to the classification system developed by Steward and Milford and femoral head fractures were classified according to the Pipkin classification. All of the hips were classified as very good, good, medium, fair, and poor according to the functional evaluation system described by Merle d'Aubigne. Statistical analysis of the results was performed. RESULTS: There were 47 male patients and 15 female patients, with ages ranging from 14 to 72 years (mean, 34.5 years). Traffic accidents constituted the leading cause of traumatic dislocation in this series (52 cases [83.9%]). Associated injuries were found in 44 cases (71%). Fifty patients were treated with closed reduction, and 12 patients were treated with open reduction. Thirty-five hips (56.5%) were reduced within 12 hours. Full weight-bearing was resumed between 2 and 10 weeks (average, 8 weeks) after injury. In follow-up periods ranging from 3.6 years to 18.4 years (mean, 9.6 years), 44 patients (71%) had very good or good to medium results. Ten patients (16.1%) developed late posttraumatic osteoarthritis of the hip, and 5 patients (9.6%) developed osteonecrosis of the femoral head. In this study, it is found that the time between injury and reduction and the associated injuries are the most important factors in long-term prognosis. CONCLUSION: We believe that good results were obtained in patients with early, stable, and accurate reductions by either closed or open methods. Concentric reduction absolutely should be confirmed by radiographs of the pelvis and, if necessary, by computed tomographic scan. The routine use of seat belts could have prevented many of these injuries. PMID- 12634534 TI - Locked-nail treatment of humeral surgical neck nonunions. AB - OBJECTIVE: The authors' purpose was to report their experience with the use of locked nails in treating humeral surgical neck nonunions. Locked nailing has the advantages of minimal soft tissue trauma, automatic fracture reduction, and stable fracture fixation. METHODS: Excluding 3 patients with severe medical illness, a total of 15 consecutive patients with 15 nonunions were treated by humeral locked nailing and bone grafting. Delay from trauma to surgery averaged 13.8 months, and 10 patients had previous operations. The average age of patients was 55.1 years; average follow-up time was 22.4 months. There were two hypertrophic and 13 atrophic nonunions. The average preoperative Neer score for shoulder joints was 53.5. All but three cases had varus deformity, with an average angle of 32.3 degrees. RESULTS: Seven nonunions were nailed with 8-mm nails and eight with 7-mm nails. With a single operation, all but one patient achieved osseous union in an average of 5.4 months. This one patient, who had a Neer score of 72, refused further surgery. At follow-up, 13 of the 15 patients had excellent or good functional recovery of shoulder joints, with an average Neer score of 86.7. Four patients had residual varus angulation averaging 15.5 degrees. For patients with solid union, the average range of shoulder motion was 152.3 degrees in flexion and 146.4 degrees in abduction. No patients had significant limitation of elbow motion. Three patients had occasional shoulder pain and no patients had evident shoulder impingement. CONCLUSION: For humeral surgical neck nonunions in which the head fragment still preserves the greater tuberosity, locked nailing with transfixing screws appeared to be a useful alternative, especially in fractures with osteoporosis, severe varus deformity, or severe soft tissue scarring. PMID- 12634535 TI - Internal fixation of displaced proximal humeral fractures with two one-third tubular plates. AB - BACKGROUND: Operative treatment of displaced proximal humeral fractures often results in poor functional outcome. We report a technique that provides improved rotational stability of the fracture and thus allows early functional treatment. METHODS: Seventy-one consecutive patients (67 +/- 18 years) with displaced fractures of the proximal humerus (two-part, n = 10; three-part, n = 41; four part, n = 20) were treated by open reduction and internal fixation using two one third tubular plates on the anterior and lateral aspects of the proximal humerus. Passive motion was started on the third postoperative day, followed by actively assisted exercises on day 6. RESULTS: Seven patients (12%) had complications (fracture redisplacement, avascular necrosis of the humeral head, frozen shoulder, subacromial impingement, and implant loosening) that required further surgical intervention. Sixty patients (85%) were available for follow-up evaluation 17 +/- 10 months after the injury. Using the Constant score, 34% of the patients had very good results, 29% had good results, 25% had fair results, and 12% had poor results. Age (< 60 years or > or = 60 years) and fracture type had no influence on functional outcome. CONCLUSION: These results demonstrate the high stability of internal fixation with two one-third tubular plates that allowed early mobilization of the shoulder in all patients and emphasize this technique as a preferred treatment option for displaced fractures of the proximal humerus. PMID- 12634536 TI - Acute complications associated with greenfield filter insertion in high-risk trauma patients. AB - BACKGROUND: Use of Greenfield filters (GFs) to prevent fatal pulmonary embolism (PE) in trauma patients is generally well accepted. Nonetheless, a surprisingly small number of trauma surgeons insert filters in their patients. Among the reasons cited is fear of complications. METHODS: We observed three femoral arteriovenous fistulae (AVF) in trauma patients who had inferior vena caval placement of filters for PE prophylaxis in one 12-month period (academic year 1999). In an effort to document the magnitude of this problem, we evaluated trauma patients who had a GF inserted in academic year 2000. RESULTS: During that year, 133 consecutive patients (8.6% of trauma admissions) received 133 GFs through a percutaneous approach. The most common isolated indications for GF insertion included closed head injuries (n = 28), multiple long bone fractures (n = 27), pelvic and acetabular fractures (n = 6), spinal cord injuries (n = 16), and vertebral fractures (n = 3). Five patients had documented deep venous thrombosis (DVT) diagnosed by duplex ultrasonography before GF placement, and 11 patients had other indications requiring a filter. There were 37 patients with more than one indication requiring filter placement. Most patients (57%) underwent preinsertion duplex scanning of their lower extremity veins; 77% of patients underwent postinsertion scanning. Filters were inserted an average of 6.8 +/- 0.6 (SE) days after trauma. No AVF were suspected clinically or detected ultrasonographically. No operative or postoperative complications occurred. DVT was observed in 30% of patients despite 92% prophylaxis; there was a 26% incidence of de novo thrombi detected. None of the patients evidenced DVT clinically. CONCLUSION: Our data indicate that complications of GF insertion for prophylaxis against PE from DVT complicating trauma patients continue to be negligible. In addition, the incidence of insertion-site thrombosis may be lower than expected. Moreover, femoral AVF is a rare complication of this procedure. PMID- 12634537 TI - Intracranial pressure response to severe head injury induced apnea and catecholamine surge. AB - BACKGROUND: Apnea and catecholamine surge have been known sequelae in the first few minutes of postexperimentally induced severe head injury for over a century. However, the intracranial pressure (ICP) response to these two pathophysiologic processes is poorly understood. METHODS: We used the rat fluid percussion head injury model to study apnea and catecholamine surge separately and in combination on measured ICP response RESULTS: The three experimental groups of apnea, hypertensive surge, and both combined revealed significantly different ICP responses with markedly elevated pressures correlating closely with mean arterial blood pressure. CONCLUSION: ICP and mean arterial blood pressure correlate closely in the first few minutes after head injury in the absence of space occupying hematomas, and may initiate pathophysiologic sequelae that can only be treated by earlier medical intervention at the scene. PMID- 12634538 TI - Metformin blunts stress-induced hyperglycemia after thermal injury. AB - BACKGROUND: Hyperglycemia is associated with detriments in immune function and impaired wound healing. The purpose of this study was to assess the effect of metformin, an oral antihyperglycemic agent approved for patients with diabetes mellitus, on glucose metabolism in severely burned patients. METHODS: Metformin was given in a double-blind, placebo-controlled fashion to 10 patients, all with burns > 60% body surface area (age, 36 +/- 4 years; weight, 92 +/- 3 kg; mean +/- SEM). After 8 days of metformin or placebo, glucose kinetics were quantitated using isotopic dilution with 6,6-d glucose and indirect calorimetry. Measurements were made during fasting; during an intravenous glucose infusion (30 micromol/kg/min); and during a hyperinsulinemic (500 mIU/m2/h), euglycemic clamp (mean plasma glucose concentration, 6.5 +/- 0.3 mmol/L). RESULTS: During fasting, metformin-treated subjects had a significantly lower rate of endogenous glucose production (met. 9.6) and glucose oxidation than placebo control subjects. With the administration of intravenous glucose, metformin treatment significantly accelerated glucose clearance, thereby attenuating hyperglycemia. During hyperinsulinemia, glucose uptake was significantly greater in metformin-treated patients. Patients receiving metformin also had a significantly higher plasma concentration of insulin. CONCLUSION: These findings suggest a potential clinical efficacy of metformin to reduce stress-induced hyperglycemia by increasing glucose clearance. This effect may be mediated by either a metformin-induced augmentation of insulin sensitivity or by increasing insulin availability. PMID- 12634539 TI - Factors affecting success of split-thickness skin grafts in the modern burn unit. AB - BACKGROUND: The use of early tangential excision or excision to fascia of burn wounds has led to the application of split-thickness skin grafts (STSGs) to a variety of graft beds, including dermis, granulation tissue, fat, and fascia. Because insufficient objective data are available on the effect of the graft bed on survival of an STSG, a 2-year review of success rates of STSGs on a variety of graft beds was completed. METHODS: The success rates of all 599 STSGs applied to dermis, granulation tissue, fat, and fascia in 233 consecutive burn patients (mean total body surface area [TBSA] burned, 14.5%) by one surgeon at a regional burn center over a 2-year period were reviewed. Data were analyzed to compare outcomes of STSGs on the graft beds listed and in low-risk versus high-risk groups of patients (TBSA burned < or = 35% and > 35%; age < or = 18 years and > 18 years; age < or = 55 years and > 55 years; and diabetes mellitus). One-way analysis of variance was used to compare results of STSGs on different graft beds, and tests were used to analyze differences in results of STSGs in low-risk versus high-risk groups (p < 0.05, significant). RESULTS: The mean success rate at 14 days for all 599 STSGs applied in the 233 patients was 90 +/- 22%. The success rate of STSG on the various surfaces ranged from 85% (fascia) to 93% (dermis; granulation), but the differences among the four graft beds were not significant. Total body surface area burned (> 35%), older age (> 55 years), and the presence of diabetes mellitus each had a significant impact on the percentage take of STSGs at 14 days after application. CONCLUSION: In the hands of an experienced burn surgeon, the recipient bed has no significant impact on the success rate of STSGs at 14 days postgrafting, except in those patients 18 years or younger, in which the mean STSG success rate was significantly greater on granulation tissue compared with fat. TBSA burned > 35%, age > 55 years, and the presence of diabetes mellitus continue to have an adverse impact on the success rate of STSGs at 14 days. PMID- 12634540 TI - Gene transfer into human keloid tissue with adeno-associated virus vector. AB - BACKGROUND Gene transfer is a new territory for clinicians. Intractable disorders might be approached in such a way. Adeno-associated virus (AAV) vector has been transfected successfully into a variety of tissues including skin. We evaluated the ability of this vector to transfer and cause expression of the reporter gene in human keloid tissue. METHODS Human keloid specimens were injected with an AAV vector encoding beta-galactosidase and incubated for 4 weeks after injection. The presence of mRNA and beta-galactosidase enzymatic activity were assayed by reverse-transcriptase polymerase chain reaction and the X-gal technique. RESULTS Gene expression shown by reverse-transcriptase polymerase chain reaction was observed in keloid tissue 4 weeks after injection, and so was the positive X-gal staining. CONCLUSION Our results showed that AAV vector could transduce human keloid tissue effectively. Replacement of the reporter gene with a functioning gene might be feasible for keloid treatment. PMID- 12634541 TI - Electron paramagnetic resonance analysis of transferrin-bound iron in animal models of blunt trauma. AB - BACKGROUND: Blood iron sequestration is known to be implicated in the systemic acute-phase response to trauma injury. The objective of the present research was to assess the effect of iron sequestration in animal models of blunt trauma by means of electron paramagnetic resonance spectroscopy of iron in complex with transferrin, a main iron-transporting protein in blood, and to correlate this effect with the extent of induced injury. METHODS: Two animal models of blunt trauma were explored in the present study. Blunt trauma in the rat model was produced by exposure of 14 animals to blast overpressure (BOP) (at peak BOP of either 86 +/- 5 kPa or 112 +/- 2 kPa) generated in a shock tube. Blunt trauma in the porcine model was produced by impact of high-speed projectiles made from a rubber-tipped, plastic composite weighing 28.64 +/- 0.12 g (mean +/- SEM, n = 8) with a length of approximately 6 cm and a diameter of approximately 4 cm. The projectiles were propelled by compressed helium onto eight animals at a velocity of 101.8 +/- 3.8 m/s (mean +/- SEM, n = 8) at the point of impact. Each experiment was accompanied by a pathology assessment using an injury scoring system developed for blunt trauma injuries to derive a severity score for whole body involvement. Amounts of transferrin-bound iron (TRF-[Fe3+]) in whole blood and blood plasma samples were measured using quantitative electron paramagnetic resonance spectroscopy. The observed alterations in the amounts of blood TRF [Fe3+] were correlated with estimated injury score ratios in each animal. RESULTS: Blunt trauma produced by either BOP exposure of rats or projectile impacts in pigs was accompanied by TRF-[Fe3+] sequestration observed in both blood and blood plasma. The amount of TRF-[Fe3+] in blood was shown to have inverse correlation with the extent of injury (Pearson r = -0.90 in the rat model and r = -0.93 in the porcine model) estimated by injury score ratios and was not dependent on location of the injury (lung, liver, spleen, or jejunum). CONCLUSION: The presented data suggest that assessment of TRF-[Fe3+] in blunt trauma can provide a good deal of information on severity of injury. The response of TRF-[Fe3+] can be considered as a potential surrogate marker of the systemic alterations in blunt trauma and, therefore, warrants further investigation in a human pilot study. PMID- 12634543 TI - Abdominal insufflation for prevention of exsanguination. AB - BACKGROUND: Currently, traumatic intra-abdominal hemorrhage continues unchecked during transport and triage, and a simple technique of prehospital hemostasis might improve outcomes. The hemostatic effect of abdominal hypertension has not been studied. PURPOSE: To examine the effect of iatrogenic abdominal insufflation on blood loss and hemodynamic performance after major abdominal vascular injury. METHODS: Following laparotomy, a 2.7 mm hole was created in the inferior vena cava of 10 anticoagulated pigs and controlled with a partially occlusive, laparoscopic vascular clamp. After abdominal closure the clamp was released and the pig was randomized to either control (n = 5) or immediate abdominal CO2 insufflation at 20 cm H2O pressure (n = 5). Lactated Ringer's solution was used as needed to maintain a mean arterial pressure of 60 mm Hg. After 15 minutes of hemorrhage and hemodynamic monitoring, the animals were killed and blood loss measured. Mean blood loss was compared between groups using the Student test, as were final values for physiologic variables. Temporal changes in physiologic parameters were compared using analysis of variance. RESULTS: Mean blood loss was reduced by 61% in insufflated pigs versus controls (695 +/- 244 versus 1764 +/- 328 cc, p < 0.001). Compared with controls, insufflated pigs had significantly higher mean arterial pressure (64 versus 25 mm Hg, p < 0.001), end-tidal CO2 (40.8 versus 17.8 mm Hg, p < 0.001), and pulmonary capillary wedge pressure (10.2 versus 5.8 mm Hg, p = 0.026) immediately before the pigs were killed. CONCLUSION: Iatrogenic abdominal insufflation significantly decreased blood loss and improved hemodynamics in a porcine model of traumatic venous hemorrhage. Iatrogenic abdominal insufflation may be useful in the prehospital management of abdominal injury. PMID- 12634542 TI - Sublethal endotoxin administration evokes super-resistance to systemic hypoxia in rats. AB - BACKGROUND: Systemic hypoxia following surgical injury modulates cytokine and catecholamine responses. Endotoxin tolerance develops after pretreatment of animals with sublethal endotoxin doses and is characterized by a reduced inflammatory cytokine response to subsequent endotoxin challenges. The administration of endotoxin also attenuates ischemic injury of rat myocardial tissue following hypoxia, a phenomenon described as cross-tolerance. The objectives of this study were: 1) to determine whether endotoxin evokes a cross tolerance to systemic hypoxia in rats; and 2) to estimate circulatory and pulmonary performance in rats with systemic hypoxia after endotoxin pretreatment. METHODS: Seventy-two hours before the experiment, Wistar rats were given an intraperitoneal injection of endotoxin at a dose of 10 microg/kg. Polyethylene catheters were inserted into the femoral vein for infusion, and into the femoral artery for blood sampling and blood pressure monitoring. Systemic hypoxia was achieved by continuous inhalation of a modified gas mixture (9% oxygen+ 91% N2) for 4 hours. Plasma TNF-alpha and IL-6 were measured by ELISA, and norepinephrine (NE) by HPLC. RESULTS: The hypoxic rats that were pretreated with saline showed a significant decrease in mean arterial blood and base excess, as compared with the normoxic rats. Endotoxin pretreatment prevented the drop in mean arterial pressure during hypoxia and reduced the decrease in base excess. Hypoxic conditions markedly stimulated TNF-alpha and IL-6 release and increased NE levels, compared to the normoxic rats. Pretreatment with endotoxin suppressed the hypoxia-induced cytokine production as well as attenuating the increase in NE levels CONCLUSIONS: In this rat hypoxia model, endotoxin pretreatment ameliorated the hypoxia-induced inflammatory response as well as suppressing the effects on arterial oxygenation, anaerobic metabolism and NE stimulation. PMID- 12634544 TI - Management of the injured patient: identification of research topics for systematic review using the delphi technique. AB - BACKGROUND: Systematic reviews of controlled clinical trials in the form of meta analyses can serve as an important guide to direct clinical practice. This study identifies the most important research questions pertaining to the acute care of the injured patient using a Web-based Delphi technique to achieve consensus of expert opinion. METHODS: Experts in trauma care from the United States and Canada (n = 68) were asked to generate structured research questions and were then required to rank these questions in order of importance and estimate the amount of research currently published. RESULTS: The questions ranking in the highest tertile are presented along with an estimate of their importance and the amount of research published using an ordinal scale. Only 9 of 16 (56%) questions had some or a substantial amount of research available on which to perform a systematic review. CONCLUSION: This study identifies the areas of trauma care in which research efforts might best be directed. In the absence of sufficient data for systematic reviews, these research topics represent important areas for the design and implementation of clinical trials. PMID- 12634545 TI - Interprosthetic fracture of the femoral shaft treated with a percutaneously inserted dynamic condylar screw: case report. PMID- 12634546 TI - Ex vivo repair and renal autotransplantation after penetrating trauma: is there an upper limit of ischemic/traumatic injury beyond which a kidney is unsalvageable? PMID- 12634547 TI - Pulmonary artery injury and cardiac tamponade after needle decompression of a suspected tension pneumothorax. PMID- 12634548 TI - Stab wounds to the neck with partial transsection of the spinal cord and penetrating injury to the esophagus. PMID- 12634549 TI - Acute pain management of patients with multiple fractured ribs. AB - BACKGROUND: Multiple rib fracture causes severe pain that can seriously compromise respiratory mechanics and exacerbate underlying lung injury and pre existing respiratory disease, predisposing to respiratory failure. The cornerstone of management is early institution of effective pain relief, the subject of this review. METHODS: A MEDLINE search was conducted for the years 1966 through and up to December 2002 for human studies written in English using the keywords "rib fractures", "analgesia", "blunt chest trauma", "thoracic injury", and "nerve block". The reference list of key articles was also searched for relevant articles. The various analgesic techniques used in patients with multiple fractured ribs were summarized. RESULTS: Analgesia could be provided using systemic opioids, transcutaneous electrical nerve stimulation or non steroidal anti-inflammatory drugs. Alternatively, regional analgesic techniques such as intercostal nerve block, epidural analgesia, intrathecal opioids, interpleural analgesia and thoracic paravertebral block have been used effectively. Although invasive, in general, regional blocks tend to be more effective than systemic opioids, and produce less systemic side effects. CONCLUSION: Based on current evidence it is difficult to recommend a single method that can be safely and effectively used for analgesia in all circumstances in patients with multiple fractured ribs. By understanding the strengths and weaknesses of each analgesic technique, the clinician can weigh the risks and benefits and individualize pain management based on the clinical setting and the extent of trauma. PMID- 12634550 TI - Missed traumatic hernia of the abdominal wall after contralateral pelvic and acetabular fracture. PMID- 12634551 TI - Disappearing thrombus on duplex resulting in a CT-proven pulmonary embolus. PMID- 12634552 TI - The value of prehospital Glasgow Coma Scale (GCS) to predict the need for patient hospitalization. PMID- 12634553 TI - Patient trauma outcomes after discontinuation of hospital-based air transport service. PMID- 12634554 TI - Strategic directions: the scoliosis research society initiatives for change. PMID- 12634555 TI - Food for thought. PMID- 12634556 TI - The effect of titanium particulate on development and maintenance of a posterolateral spinal arthrodesis: an in vivo rabbit model. AB - STUDY DESIGN: The current study was undertaken to determine if the presence of titanium wear particulate deleteriously influences early osseointegration of posterolateral bone graft or disrupts an established posterolateral fusion mass. OBJECTIVES: Using an in vivo animal model to evaluate the effect(s) of titanium wear particulate on a posterolateral spinal arthrodesis based on serologic, histologic, and immunocytochemical analyses. SUMMARY OF BACKGROUND DATA: The effect of unintended wear particulate resulting from micromotion between the interconnection mechanisms in spinal instrumentation remains a clinical concern. METHODS: Thirty-four New Zealand White rabbits were randomized into two groups based on postoperative time periods of 2 months (Group 1, n = 14) and 4 months (Group 2, n = 20). Group 1 underwent a posterolateral arthrodesis at L5-L6 using tricortical iliac autograft or tricortical iliac autograft + titanium particulate. Group 2 received iliac autograft at the initial surgery and were reoperated on after 8 weeks and treated with posterolateral arthrodesis exposure alone or titanium particulate. Postoperative analysis included serologic quantification of systemic cytokines. Postmortem microradiographic, immunocytochemical, and histopathologic assessment of the intertransverse fusion mass quantified the extent of osteolysis, local pro-inflammatory cytokines, osteoclasts, and inflammatory infiltrates. RESULTS: Serologic analysis of systemic cytokines indicated no significant differences in cytokine levels (P > 0.05) between the titanium or autograft treatments. Immunocytochemistry indicated increased levels of local cytokines (tumor necrosis factor-alpha) at the titanium treated posterolateral arthrodesis sites at both time periods (P < 0.05). Osteoclast cell counts and regions of osteolytic resorption lacunas were higher in the titanium-treated versus autograft-alone groups (P < 0.05), and the extent of cellular apoptosis was markedly higher in the titanium-treated sites at both time intervals. Electron microscopy indicated definitive evidence of phagocytized titanium particles and foci of local, chronic, inflammatory changes in the titanium-treated sites. CONCLUSION: Titanium particulate debris introduced at the level of a spinal arthrodesis elicits a cytokine-mediated particulate-induced response favoring pro-inflammatory infiltrates, increased expression of intracellular tumor necrosis factor-alpha, increased osteoclastic activity, and cellular apoptosis. The presence of titanium particulate debris, secondary to motion between spinal implants, may serve as the impetus for late-onset inflammatory-infectious complications and long-term osteolysis of an established posterolateral fusion mass in the clinical setting. PMID- 12634557 TI - Dowel fibular strut grafts for high-grade dysplastic isthmic spondylolisthesis. AB - STUDY DESIGN: This is a clinical study that examines the results of partial reduction and fibular dowel graft placement for high-grade isthmic spondylolisthesis. OBJECTIVES: To demonstrate the efficacy of partial reduction and fibular dowel graft placement in the treatment of high-grade isthmic spondylolisthesis. SUMMARY OF BACKGROUND DATA: Previous literature has demonstrated difficulty in treating high-grade isthmic spondylolisthesis both with high rates of pseudarthrosis as well as neurologic complications if a complete reduction is attempted. There are no published data examining partial reduction with dowel graft placement. METHODS: Seventeen consecutive patients (mean age 20.3 years) with high-grade isthmic spondylolisthesis who were treated with posterior fusion and fibular strut grafts were studied (mean follow-up 4.6 years). Radiographs were reviewed at preoperative, immediate (within 3 months) postoperative, and ultimate (>2 years) follow-up. Parameters measured included lumbar lordosis, slip angle, Meyerding-Newman scores, and pelvic incidence. The anterior and posterior fusions were graded on a I-IV scale, and the implants (if used) were examined for failure. Clinical outcomes were measured with Oswestry and Scoliosis Research Society outcomes tools. RESULTS: There were 17 patients treated: 10 primary and 7 revision patients. All patients had posterior fusion with fibular dowel grafts (11 allograft, 6 autograft). Meyerding grade improved 1.3 grades and slip angle improved 14 degrees with no loss of correction at ultimate follow-up. Sixteen of 17 patients had solid fusions on ultimate follow up. Clinical evaluation with Scoliosis Research Society and Oswestry tools showed high patient function and satisfaction. Complications included one case of a broken strut in a revision patient; this was then revised to an instrumented circumferential fusion. There were no cases of deep or superficial infection. There were no neurologic deficits at ultimate follow-up. CONCLUSION: Fibular strut grafting is a useful surgical adjunct in high-grade spondylolisthesis that is partially reduced. Clinical and radiographic outcomes were satisfactory. Our experience shows that there is no significant difference between allograft and autograft. All struts healed and remodeled by the ultimate follow-up, and there was only one instance of fibula fracture. PMID- 12634558 TI - Treatment of spinal fractures in children and adolescents: long-term results in 44 patients. AB - STUDY DESIGN: The authors examined a case series of patients younger than 16 years who had sustained a traumatic spine injury. OBJECTIVES: To evaluate clinical and radiologic findings and the effectiveness of conservative versus surgical treatment at long-term follow-up. SUMMARY OF BACKGROUND DATA: Although injuries to children have received increasing coverage in the literature over the last several years, few reports have focused on the long-term results of conservative versus surgical treatment. METHODS: Forty-four patients who had sustained a traumatic spine injury at the average age of 14 years (range 3-16 years) were clinically and radiographically reviewed. The fractures were separated into three groups: stable (n = 20) and unstable (n = 13) injuries without cord lesion and fractures with spinal cord lesion (n = 11). Mean follow up was 18 years (range 9-23 years). RESULTS: Conservative treatment was successful in all stable fractures, whereas it failed in the unstable injuries. The surgical treatment stabilized without significant deformity in five of the seven unstable fractures. Of 11 with spinal cord injuries, the 4 children conservatively treated developed a severely progressive, paralytic scoliosis. Only three of the seven surgically treated patients were stabilized without any deformity at follow-up. CONCLUSION: In children and adolescents, conservative treatment is an available option for stable fractures without neurologic lesion. Early surgical treatment (instrumentation and fusion) is mandatory for unstable fractures and injuries associated with spinal cord lesion. In children, a traumatic spinal cord lesion may develop a deformity that is mainly scoliotic, kyphotic, or lordotic in >90% of the cases. PMID- 12634559 TI - Spinal deformity in chondrodysplasia punctata. AB - BACKGROUND DATA: Our purpose was to describe the deformities, natural history, and course of treatment in chondrodysplasia punctata, a skeletal dysplasia characterized by stippled epiphysis in infancy, short stature, and progressive kyphoscoliosis. STUDY DESIGN: We identified 20 patients (15 female, 5 male) with chondrodysplasia punctata and spinal abnormalities. RESULTS: The spinal deformity was rarely present in the neonatal radiographs but typically appeared during the first year. We identified three patterns of spinal deformity. The first was a C2 dentocentral synchondrosis separation, which was successfully fused. Scoliosis had two patterns: one slowly progressive without dysplastic vertebrae or significant kyphosis (two patients) and the other rapidly progressive with dysplastic vertebral bodies often appearing similar to a congenital kyphoscoliosis (17 patients). Follow-up averaged 7.9 years (range 2-20 years). The slowly progressive type was fused successfully with standard posterior techniques. The dysplastic type, however, required on average 2.5 procedures, including anteroposterior fusions, repeat fusions for pseudarthrosis repair, extension of the fusions for progression beyond the curves, and osteotomies for progression through the fusion mass. Unlike congenital deformities, they did not respond well to hemiepiphysiodesis. CONCLUSION: Three types of spinal deformities were identified in children with chondrodysplasia punctata, including cervical bony disruption, a slowly progressive, nondysplastic scoliosis responding well to standard fusion techniques and a dysplastic kyphoscoliosis, which is rapidly progressive and resistant to fusion. The best results in dysplastic curves are obtained with an anterior strut graft and a posterior fusion. The patients must be observed long-term for further vertebral dysplasia and progressive kyphoscoliosis. PMID- 12634560 TI - Spine deformity correction in Marfan syndrome. AB - STUDY DESIGN: Retrospective review of records and radiographs with postoperative questionnaire of patients with Marfan syndrome receiving surgical correction for spinal deformity. OBJECTIVE: To characterize and analyze the challenges of surgical correction for spinal deformity in Marfan syndrome and to present recommendations that may reduce morbidity and increase success for these procedures. SUMMARY OF BACKGROUND DATA: Spinal deformities are common in Marfan syndrome and usually refractory to conservative management. The few, smaller studies of surgical intervention reported on earlier surgical techniques but suggested increased complication rates. METHODS: Records and radiographs of 39 patients with confirmed Marfan syndrome who underwent surgical management at either of two institutions for primary scoliosis (n = 26), kyphosis (n = 7), or deformity secondary to previous surgery elsewhere (n = 6) were reviewed. Presentation features, complications, and therapeutic results were analyzed. Low back outcome scores were generated from a questionnaire given to patients after surgery. RESULTS: Increased blood loss and rates of infection (10%), dural tear (8%), instrumentation fixation failure (21%), pseudarthrosis (10%), and coronal (8%) and sagittal (21%) curve decompensation were noted. Infection was often associated with dural tear and decompensation with extreme correction. One patient died of valvular insufficiency 11 weeks after surgery. CONCLUSION: The cardiopulmonary condition of patients with Marfan syndrome should be evaluated and planned for before surgery. Preoperative computed tomograph to assess bony adequacy for fixation and magnetic resonance imaging to evaluate dural ectasia are indicated. Attention paid to the sagittal profile, extension of fusion to vertebrae neutral and stable in both planes, minimization of soft tissue dissection, and avoidance of extreme correction may prevent curve decompensation. PMID- 12634561 TI - Correlation of radiographic, clinical, and patient assessment of shoulder balance following fusion versus nonfusion of the proximal thoracic curve in adolescent idiopathic scoliosis. AB - STUDY DESIGN: Retrospective clinical, radiographic, and patient outcome review of surgically treated adolescent idiopathic scoliosis. OBJECTIVES: To correlate radiographic and clinical features of shoulder balance and the proximal thoracic curve with patient satisfaction outcomes at a minimum 2-year follow-up. SUMMARY OF BACKGROUND DATA: Traditionally, radiographic features of a structural proximal thoracic curve have been T1 tilt, proximal thoracic Cobb angle, and proximal thoracic side-bending Cobb; however, these do not always correlate with clinical shoulder balance. METHODS: A total of 112 patients (single surgeon) with adolescent idiopathic scoliosis and a proximal thoracic curve >or=20 degrees (average 32 degrees, range 20-78 degrees) were evaluated in terms of shoulder balance and curve flexibility/correction. Four groups were analyzed: Group 1, posterior spinal fusion to T2 (proximal thoracic curve included, n = 24); Group 2, posterior spinal fusion to T3 (proximal thoracic curve partially included, n = 23); Group 3, posterior spinal fusion to T4 or T5 (proximal thoracic curve not included, n = 21); and Group 4, anterior spinal fusion to T4 or below (proximal thoracic not included, n = 44). Proximal thoracic, main thoracic, and thoracolumbar-lumbar upright coronal, side-bending, and sagittal Cobb measurements were assessed before surgery, 1 week after surgery, and at a minimum 2-year postoperative follow-up (average 3.8 years, range 2.0-7.6 years). In addition to T1 tilt, clavicle angle (intersection of a horizontal line and the tangential line connecting the highest two points of each clavicle), coracoid height difference, trapezius length (horizontal distance of the T2 pedicle to second rib-clavicle intersection), first rib-clavicle height difference (vertical distance of first rib apex to superior clavicle), and proximal thoracic, main thoracic, and thoracolumbar-lumbar apical vertical translation were determined. Shoulder asymmetry as measured by the radiographic soft tissue shadow was graded as balanced (<1 cm), slight (1-2 cm), moderate (2-3 cm), or significant (>3 cm). A postoperative patient questionnaire addressed shoulder balance and overall appearance at most recent follow-up. RESULTS: The four groups were found to be statistically equivalent in terms of preoperative proximal thoracic curve (P = 0.4146), proximal thoracic side-bending Cobb (P = 0.2199), main thoracic curve (P = 0.6999), and main thoracic side-bending curves (P = 0.7307). Radiographic: Preoperative proximal thoracic measurements correlating with postoperative shoulder balance (P < 0.05) included the clavicle angle (three of four groups with a trend toward statistical significance in the fourth group, P = 0.07) and coracoid height (two of four groups). No other measurement, including T1 tilt and proximal thoracic side-bending Cobb, correlated in more than one group. Proximal thoracic curve correction was greatest in Group 1 (posterior spinal fusion to T2; average 12 degrees) and Group 4 (anterior spinal fusion to T4 or below; average 12 degrees). Clinical: Shoulder balance improved in all four groups (range 0.38 1.00 grades). There was no difference in shoulder balance between groups (P = 0.2723). Patient assessment: All four groups also reported improvement in self perceived shoulder balance (63% up to one grade, 37% over two-grade improvement), whereas no patient reported worsening of shoulder balance. There was no significant difference in patient outcomes between the four groups (P = 0.3654). CONCLUSION: The clavicle angle, not T1 tilt, upright proximal thoracic, or side bending proximal thoracic Cobb, provided the best preoperative radiographic prediction of postoperative shoulder balance. In each of the four groups, postoperative shoulder balance and clinical appearance also improved and correlated with patient postoperative assessments. PMID- 12634562 TI - Radiographic markers in spondyloptosis: implications for spondylolisthesis progression. AB - STUDY DESIGN: Radiographic analysis of spinopelvic morphology and posterior element dysplasia in spondyloptosis. SUMMARY OF BACKGROUND DATA: Spondylolisthesis treatment protocols are based on age, symptomatology, and slippage degree. Spinopelvic morphology and dysplasia can determine progression. Frequency of two denominators of high-grade spondylolisthesis-degree of dysplasia and spinopelvic morphology-is unknown. OBJECTIVES: To determine common radiographic denominators of spondyloptosis-degree of posterior bony hook dysplasia and spinopelvic morphology-as prognostic factors for spondylolisthesis progression. METHODS: Patients with spondyloptosis were reviewed. Bony dysplasia at lumbosacral junction was graded. Pelvic incidence and sacral kyphosis were measured. RESULTS: A total of 53 patients had a mean sacral kyphosis of 56 degrees and pelvic incidence of 76 degrees; 62% of patients had posterior element dysplasia. CONCLUSION: Prognostic factors for spondylolisthesis progression, such as percent of slippage, do not identify lower-grade slips at risk for progression. Progression is linked to increased shear stress across the lumbosacral junction and inability to resist it. Increased stress is related to increased verticality of the lumbosacral joint, which is individually predetermined by pelvic incidence and sacral anatomy. Pelvic incidence is fundamental in determining sagittal spine curvature required for economic spinopelvic balance. Pelvic incidence is independent of adaptive changes in higher-grade spondylolisthesis. Pelvic incidence in our spondyloptosis series (76 degrees) is higher than in normal (48.2-53.2 degrees ) and low-grade spondylolisthesis (64.5 degrees). Posterior element dysplasia decreases mechanical resistance to lumbosacral shear stress. Incidence of dysplasia in our series (62%) is higher than that reported in low-grade spondylolisthesis. Analysis of pelvic incidence and posterior element dysplasia may aid in estimation of risk for progression of spondylolisthesis. PMID- 12634563 TI - Correlation of pelvic incidence with low- and high-grade isthmic spondylolisthesis. AB - PURPOSE: The development of isthmic spondylolisthesis is influenced by forces across the lumbosacral region of the spine. Pelvic incidence is a radiographic parameter that has been shown to be an independent parameter that influences both sagittal spinal balance and pelvic orientation. Our hypothesis then is that there is a positive correlation between pelvic incidence and spondylolisthesis. STUDY DESIGN: A radiographic analysis of cases with spondylolisthesis. OBJECTIVES: To try to assess the correlation between pelvic incidence in both low-grade and high grade spondylolisthesis in both a pediatric and an adult population. SUMMARY OF BACKGROUND DATA: The concept of pelvic incidence has been introduced into the literature. Its exact association with spondylolisthesis has not yet been clarified. METHODS: Forty patients with spondylolisthesis were identified and divided into two groups: low-grade (Meyerding I-II) and high-grade (Meyerding III and higher). Radiographic parameters measured included lumbar sagittal alignment (T12-S1), sacral inclination, slip angle, and pelvic incidence. The spondylolisthesis was classified according to the Meyerding-Newman classifications and the slip angle. Radiographic measurements were also done in two control groups; there were 20 pediatric and 20 adult controls (mean age 11.8 years and 60.0 years, respectively). Unpaired t test analysis and Pearson correlation analysis were then done. RESULTS: Mean pelvic incidence was 47.4 degrees in the pediatric control group, 57 degrees in the adult control group, 68.5 degrees in the low-grade isthmic spondylolisthesis group, and 79.0 degrees in the high-grade isthmic spondylolisthesis group. Pelvic incidence was found to be significantly higher in the high- and low-grade spondylolisthesis groups compared with both control groups (P = 0.0001). Pelvic incidence was significantly higher in the high-grade isthmic spondylolisthesis group than in the low-grade isthmic spondylolisthesis group (P = 0.007). A significant correlation existed between pelvic incidence and Meyerding-Newman scores (P = 0.03). CONCLUSIONS: Pelvic incidence was significantly higher in patients with low- and high-grade isthmic spondylolisthesis as compared with controls and had significant correlation with the Meyerding-Newman grades (P = 0.03). PMID- 12634564 TI - Can triggered electromyograph thresholds predict safe thoracic pedicle screw placement? AB - STUDY DESIGN: A prospective clinical study of thoracic pedicle screws monitored with triggered electromyographic testing. OBJECTIVE: To evaluate the sensitivity of recording rectus abdominis triggered electromyographs to assess thoracic screw placement. SUMMARY OF BACKGROUND DATA: Triggered electromyographic testing from lower extremity myotomes has identified medially placed lumbar pedicle screws. Higher thresholds indicate intraosseous placement because of increased resistance to current flow. Lower thresholds correspond to compromised pedicles with potential for nerve impingement. No clinical study has correlated an identical technique with rectus muscle recordings, which are innervated from T6 to T12. METHODS: A total of 677 thoracic screws were placed in 92 consecutive patients. Screws placed from T6 and T12 were evaluated using an ascending method of stimulation until a compound muscle action potential was obtained from the rectus abdominis. Threshold values were compared both in absolute terms and also in relation to other intrapatient values. RESULTS: Screws were separated into three groups: Group A (n = 650 screws) had thresholds >6.0 mA and intraosseus placement. Group B (n = 21) had thresholds <6.0 mA but an intact medial pedicle border on reexamination and radiographic confirmation. Group C (n = 6) had thresholds <6.0 mA and medial wall perforations confirmed by tactile and/or visual inspection. Thus, 3.9% (27 of 677) of all screws had thresholds <6.0 mA. Only 22% (6 of 27) had medial perforation. Group B screws averaged a 54% decrease from the mean as compared with a 69% decrease for Group C screws (P = 0.0160). There were no postoperative neurologic deficits or radicular chest wall complaints. CONCLUSION: To assess thoracic pedicle screw placement, triggered electromyographic thresholds <6.0 mA, coupled with values 60-65% decreased from the mean of all other thresholds in a given patient, should alert the surgeon to suspect a medial pedicle wall breach. PMID- 12634566 TI - Evaluation of ventilatory efficiency during exercise in patients with idiopathic scoliosis undergoing spinal fusion. AB - STUDY DESIGN: A prospective evaluation of ventilatory function following spinal fusion in adolescent idiopathic scoliosis. OBJECTIVES: To prospectively evaluate pulmonary function, maximal oxygen uptake, and ventilatory efficiency during exercise in patients with adolescent idiopathic scoliosis before surgery and a minimum of 2 years postoperation. SUMMARY OF BACKGROUND DATA: For reasons that are unclear, patients with untreated adolescent idiopathic scoliosis tend to avoid aerobic exercise. Their reluctance may be the result of low ventilatory efficiency, as they often approach their ventilatory ceiling at maximum oxygen uptake despite forced vital capacities that are near normal. This inefficiency of ventilation with exercise may explain the reluctance of patients with scoliosis to pursue aerobic fitness. No study has evaluated the effect spinal fusion has on the ventilatory function of patients with scoliosis during exercise. METHODS: Forty-two patients with adolescent idiopathic scoliosis (36 female and 6 male) at an average age of 14 +/- 3 years (range 10-18 years) underwent spinal fusion. Twenty patients underwent a posterior spinal fusion alone, 20 an anterior spinal fusion alone, and 2 an anterior spinal fusion and posterior spinal fusion. The average Cobb measurement was 55 degrees (range 40-85 degrees). Pulmonary function values (forced vital capacity, total lung capacity, maximum voluntary ventilation), maximum oxygen uptake (VO2max), and ventilatory efficiency were obtained before surgery and a minimum of 2 years postoperation. RESULTS: For all patients, forced vital capacity percent predicted decreased from 88.1% to 81.4% (P < 0.0001). Total lung capacity also declined from 90.5% to 88.5% but was not statistically significant (P = 0.189). Percent predicted maximum oxygen uptake (VO2max) declined from 93.6% to 85.1% (P = 0.00029). Ventilatory efficiency, as measured by VEmax/maximum voluntary ventilation, improved from 0.76 to 0.68 (P = 0.005), whereas measured by VEmax/FEV1 x 40 was unchanged from 0.69 to 0.70 (P = 0.172) postoperation. The choice of operative approach [anterior (n = 20) versus posterior (n = 20)] or whether rib graft was harvested (n = 33) versus iliac crest graft (n = 7) did not change these results. CONCLUSION: Improvement in ventilatory efficiency during exercise does not occur in the majority of patients with adolescent idiopathic scoliosis following spinal fusion and thus cannot be relied on to foster increases in aerobic activity. PMID- 12634565 TI - A multicenter study analyzing the relationship of a standardized radiographic scoring system of adolescent idiopathic scoliosis and the Scoliosis Research Society outcomes instrument. AB - STUDY DESIGN: A multicenter study examining the association between radiographic and outcomes measures in adolescent idiopathic scoliosis. OBJECTIVES: To evaluate the association between an objective radiographic scoring system and patient quality of life measures as determined by the Scoliosis Research Society outcomes instrument. SUMMARY OF BACKGROUND DATA: Although surgical correction of scoliosis has been reported to be positively correlated with patient outcomes, studies to date have been unable to demonstrate an association between radiographic measures of deformity and outcomes measures in patients with adolescent idiopathic scoliosis. METHODS: A standardized radiographic deformity scoring system and the Scoliosis Research Society outcome tool were used prospectively in seven scoliosis centers to collect data on patients with adolescent idiopathic scoliosis. A total of 354 data points for 265 patients consisting of those with nonoperative or preoperative curves >or=10 degrees, as well as those with surgically treated curves, were analyzed. Correlation analysis was performed to identify significant relationships between any of the radiographic measures, the Harms Study Group radiographic deformity scores (total, sagittal, coronal), and the seven Scoliosis Research Society outcome domains (Total Pain, General Self Image, General Function, Activity, Postoperative Self-Image, Postoperative Function, and Satisfaction) as well as Scoliosis Research Society outcomes instrument total scores. Radiographic measures that were identified as significantly correlated with Scoliosis Research Society outcome scores were then entered into a stepwise regression analysis. RESULTS: The coronal measures of thoracic curve and lumbar curve magnitude were found to be significantly correlated with the Total Pain, General Self-Image, and total Scoliosis Research Society scores (P < 0.0001). The thoracic and upper thoracic curve magnitudes were also correlated with General Function (P < 0.002). The "coronal" subscore as well as the "total" score of the Harms Study Group radiographic scoring system were also significantly correlated with these Scoliosis Research Society domain and total scores. No radiographic measures taken after surgery were significantly correlated with the postoperative domains of the Scoliosis Research Society outcomes instrument. Stepwise regression analysis of these radiographic measures as predictors of Scoliosis Research Society scores resulted in adjusted R2 values of 0.03-0.07 (P < 0.0001). Although these results show that a significant association exists between the radiographic Cobb angle measure of the scoliosis and the Scoliosis Research Society outcomes scores, the low R2 values indicate that variables other than the radiographic appearance of the deformity (e.g., psychosocial, functional) must also be affecting these scores. CONCLUSION: The Cobb angle measure of the major deformity has a small, but statistically significant, correlation with the reported Total Pain, General Self-Image, and General Function as measured by the Scoliosis Research Society outcomes instrument. None of the radiographic measures in this population correlated with postoperative domain scores of the Scoliosis Research Society outcomes tool. PMID- 12634567 TI - A multicenter study of the outcomes of the surgical treatment of adolescent idiopathic scoliosis using the Scoliosis Research Society (SRS) outcome instrument. AB - STUDY DESIGN: A multicenter study of the outcomes of the surgical treatment of adolescent idiopathic scoliosis using the Scoliosis Research Society Questionnaire (SRS 24). OBJECTIVE: To evaluate the patient based outcome of the surgical treatment of adolescent idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: A paucity of information exists with respect to patient measures of outcome regarding the surgical treatment of adolescent idiopathic scoliosis. To our knowledge, no prospective outcome study on this topic thus far exists. METHODS: Using the SRS 24 questionnaire, seven scoliosis centers agreed to prospectively assess outcome for surgically treated patients with adolescent idiopathic scoliosis. Data were collected before surgery and at 24 months after surgery. Data were analyzed using paired and independent samples t test for all seven SRS 24 questionnaire domains (Pain, General Self-Image, Postoperative Self-Image, Postoperative Function, Function From Back Condition, General Level of Activity, and Satisfaction) using Statistical Package for Social Science. The domains were analyzed with respect to the total cohort, gender, curve magnitude, and type of surgery using independent-samples t tests. RESULTS: A total of 242 patients were included in our analysis. A baseline preoperative pain level of 3.68 of 5 was found. This improved to 4.63 after surgery, representing an improvement of 0.95 points. Surgical intervention was associated with improving outcome when compared with preoperative status. Pain, General Self-Image, Function From Back Condition, and Level of Activity all demonstrated statistically significant improvement as compared with preoperative status (P < 0. 001). Overall, patients were highly satisfied with the results of surgery. CONCLUSION: Preoperative pain exists in our adolescent scoliosis population. Pain scores were improved in our study population at the 2-year postsurgical follow-up. Statistically significant improvements were likewise seen in the General Self-Image, Function From Back Condition, and Level of Activity domains. The present study demonstrates the ability of surgery to improve the outcome of patients afflicted with adolescent idiopathic scoliosis. PMID- 12634568 TI - The Pediatric Outcomes Data Collection Instrument (PODCI) and functional assessment in patients with adolescent or juvenile idiopathic scoliosis and congenital scoliosis or kyphosis. AB - STUDY DESIGN: An initial examination of functional assessment scores in scoliosis and kyphosis populations. OBJECTIVE: Examination of scores from the Pediatric Outcomes Data Collection Instrument for patients with idiopathic scoliosis, congenital scoliosis, and congenital kyphosis, comparing scores with those of children without orthopedic disabilities. SUMMARY OF BACKGROUND DATA: Little information has been presented regarding performance of scoliosis patients on the Pediatric Outcomes Data Collection Instrument. METHODS: A total of 102 patients with adolescent idiopathic scoliosis, 47 with congenital scoliosis without kyphosis, and 9 with congenital kyphosis completed the Pediatric Outcomes Data Collection Instrument. Responses were compared with those from a "normal" population. Subgroup analyses were performed for patients with adolescent idiopathic scoliosis. A P value <0.05 was considered statistically significant. RESULTS: Scores in Transfers, Sports, Comfort, and Happiness were significantly lower than "normal" in adolescent idiopathic scoliosis. In congenital scoliosis without kyphosis, scores in all categories except Happiness were significantly lower than "normal." All category scores were significantly lower than "normal" in congenital kyphosis. In adolescent idiopathic scoliosis, age and curve location did not influence Comfort scores. Comfort scores were significantly lower than "normal" for all curve locations and for all ranges of Cobb angle. Happiness scores were significantly lower in adolescent idiopathic scoliosis patients with Cobb angles >50 degrees who had not received surgery when compared with either patients who had received surgery or patients with Cobb angles <50 degrees. CONCLUSION: These findings provide some normative values for the Pediatric Outcomes Data Collection Instrument for three specific diagnoses. Patients with adolescent idiopathic scoliosis, congenital scoliosis, and congenital kyphosis gave responses significantly different from "normal" children. Pain appeared to be a common finding with these diagnoses. PMID- 12634570 TI - Correction of thoracic adolescent idiopathic scoliosis with segmental hooks, rods, and Wisconsin wires posteriorly: it's bad and obsolete, correct? AB - STUDY DESIGN: Forty-four consecutive thoracic idiopathic adolescent curves surgically treated between 1995 and 1999 at one institution were analyzed. All were thoracic curves; there were no lumbar curves, double major curves, or triple major curves. OBJECTIVES: To study the clinical and radiographic results for these 44 patients. SUMMARY OF BACKGROUND DATA: There is currently ongoing discussion regarding the "best treatment" for right thoracic idiopathic scoliosis. METHODS: Patients were administered preoperative and postoperative Scoliosis Research Society questionnaires. Radiographs were studied for coronal correction, sagittal correction, junctional deformities two segments above/two segments below, fusion status, loss of correction status, complications, and imbalance of curves above and below. Radiographic analysis was by two independent observers. All patients were treated with bilateral 5.5-mm CD Horizon (M-10) instrumentation. Corrective forces were translational and in situ contouring. An average of 10 hooks and 4 Wisconsin wires were used for an average of 14 fixation points per 11 segments. Follow-up was minimum 2 years to maximum 5 years, with an average of 3-year, 10-month follow-up. RESULTS: All were out of bed on postoperative day 1; none was braced after surgery. The average preoperative thoracic curve was 56 degrees; the average ultimate postoperative curve was 31 degrees (P = 0.0001). In the sagittal plane T5-T12 averaged 22 degrees preoperation and 20 degrees postoperation. For the complete study group (n = 44), the Cobb measurement (P = 0.001) and apical vertebral translation of the lumbar curve (P = 0.001) below improved. For the false double major curve (n = 11) patterns, the unfused lumbar curve averaged preoperation 46 degrees and postoperation 27 degrees (P = 0.0001). There were no neurologic deficits, no wound infections, and no respiratory complications. In regards to the Scoliosis Research Society questionnaires, significant (P < 0.05) changes preoperative to ultimate postoperative were found for questions 3, 5, 11, and 14. CONCLUSION: "Satisfactory" results were obtained in 44 patients treated with this technique. The complication rate was low and the fusion rate was high. Posterior treatment as described above produces reasonable and dependable results. PMID- 12634571 TI - The use of Epoetin alfa in complex spine deformity surgery. AB - STUDY DESIGN: A prospective, randomized trial comparing Epoetin alfa (Procrit) with placebo saline injection to determine effectiveness in increasing erythropoietic recovery in complex spine deformity surgery. OBJECTIVES: To determine if Epoetin alfa can allow preoperative autologous donation completion more effectively and reduce perioperative homologous blood transfusion. SUMMARY OF BACKGROUND DATA: The use of Epoetin alfa has been studied, primarily in the arthroplasty literature, for its effectiveness in decreasing transfusion requirements and increasing hemoglobin levels. It has not been studied in patients undergoing complex spine deformity surgery. METHODS: A total of 48 patients were prospectively randomized into an Epoetin alfa group and a control group. All patients attempted to donate 4 units of preoperative autologous donation at weekly intervals; 40,000 units of Epoetin alfa were injected subcutaneously at the time of preoperative autologous donation in the Epoetin alfa group. Hematocrit levels were recorded weekly during the donation process and daily in the preoperative period. RESULTS: Preoperative autologous donation was completed more effectively in the patients receiving Epoetin alfa. Epoetin alfa resulted in statistically higher hematocrit levels during preoperative autologous donation and perioperatively (P < 0.005). Homologous transfusion was decreased by 2.4 units and hospital stay was 1.8 days shorter in patients receiving Epoetin alfa. CONCLUSION: Patients who received Epoetin alfa were able to complete preoperative autologous donation more effectively, increase erythropoietic recovery, decrease homologous transfusion requirements, and had shorter hospital stays. PMID- 12634572 TI - Pelvic or lumbar fixation for the surgical management of scoliosis in duchenne muscular dystrophy. AB - STUDY DESIGN: This retrospective study evaluates two groups of patients with scoliosis and Duchenne muscular dystrophy, treated with two different surgical stabilization methods. OBJECTIVE: To determine whether fixation to the sacropelvis is always necessary for adequate stabilization of scoliosis in Duchenne muscular dystrophy. SUMMARY OF BACKGROUND DATA: Pelvic fixation is generally recommended for scoliosis in Duchenne muscular dystrophy. Recent studies describe a more selective approach toward lumbar or pelvic fixation. Pelvic fixation is reserved for larger curves and established pelvic tilt. METHODS: Fifty cases of Duchenne muscular dystrophy, operated in two different centers and followed up for a minimum of 3 years, were reviewed. In the first group (Oswestry), 31 patients had fixation to the pelvis, using standard Luque instrumentation and pelvic fixation. The Galveston technique was used in 9 cases and L-rod configuration in 22 cases. In the second group (Nottingham), 19 cases had fixation to L5 using pedicle screws in the lumbar spine and sublaminar wires in the thoracic spine. These cases were operated on early, usually shortly after becoming wheelchair dependent. RESULTS: In the pelvic fixation group, the mean age at the time of surgery was 14 years, and forced vital capacity was 44%. The mean Cobb angle and pelvic obliquity were 48 degrees and 19.8 degrees at the time of surgery, 16.7 degrees and 7.2 degrees immediately after surgery, and 22 degrees and 11.6 degrees at the final follow-up (mean 4.6 years), respectively. The mean blood loss was 4.1 L, and the average hospital stay was 17 days. There were five major complications, including a deep wound infection in one case, revision of instrumentation prominence at the proximal end in two cases, and loosening of pelvic fixation in two cases. In the lumbar fixation group, the mean age at the time of surgery was 11.7 years, and forced vital capacity was 58%. The mean Cobb angle and pelvic obliquity were 19.8 degrees and 9 degrees at the time of surgery, 3.2 degrees and 2.2 degrees immediately after surgery, and 5.2 degrees and 2.9 degrees at the final follow-up (mean 3.5 years), respectively. The mean estimated blood loss (3.3 L) and mean hospital stay (7.7 days) were much less compared with the pelvic fixation group. Pelvic obliquity was corrected and maintained below 10 degrees in all but two cases, who had an initial pelvic obliquity exceeding 20 degrees. One patient had instrumentation failure at the proximal end, and one had a deep wound infection. CONCLUSION: Lumbar fixation to L5 is adequate if the surgery is performed early, soon after becoming wheelchair bound, and with smaller curves and minimal pelvic obliquity. Use of pedicle screws in lumbar spine provides a solid foundation to maintain the correction over the period of relatively short life expectancy of these children. Pelvic fixation may be necessary in older children, who have larger curves and established pelvic obliquity. In the presence of deteriorating lung function, this is associated with a greater morbidity and higher complication rate. PMID- 12634573 TI - The relation between vertebral endplate shape and lumbar disc herniations. PMID- 12634574 TI - Posterolateral endoscopic excision for lumbar disc herniation: surgical technique, outcome, and complications in 307 consecutive cases. PMID- 12634575 TI - Clinical practice guideline for the physiotherapy of patients with whiplash associated disorders. PMID- 12634576 TI - Purely epidural spinal meningioma mimicking metastatic tumor: case report and review of the literature. AB - STUDY DESIGN: Case report. OBJECTIVES: To report a rare case of purely extradural spinal meningioma in a patient with previous history of malignant disease and to discuss the potential pitfalls in differential diagnosis and treatment of intraspinal extramedullary masses. SUMMARY AND BACKGROUND DATA: Spinal meningiomas located purely in the extradural space are rare, and they may easily be confused with malignant neoplasms that are much more common in this location. METHODS: An asymptomatic intraspinal extramedullary mass in the thoracolumbar region was detected by magnetic resonance imaging in a 75-year-old woman with previous history of malignant disease. RESULTS: The purely extradural lesion was completely excised via a dorsal approach. The intraoperative and postoperative histologic examination revealed a meningioma. The patient is symptom free 18 months after surgery without radiologic recurrence. CONCLUSION: Extradural spinal meningiomas may mimic metastatic disease. Attention needs to be drawn to this dangerous preoperative and intraoperative misinterpretation, which may easily entail an inadequate therapy. Intraoperative histology is mandatory for optimal surgical decision-making. PMID- 12634577 TI - Unilateral calf hypertrophy seen in lumbosacral stenosis: case report and review of the literature. AB - STUDY DESIGN: A case report of a patient with neurogenic unilateral calf hypertrophy and review of the literature are reported. OBJECTIVES: To provide further evidence that S1 radiculopathy is predisposed to develop neurogenic muscle hypertrophy. SUMMARY OF BACKGROUND DATA: Calf hypertrophy, specifically hypertrophy of the gastrocnemius muscle, is a rare but recognized presentation of S1 and less commonly L5 radiculopathies. The pathophysiology of this is incompletely understood. METHODS: We present a 59-year-old patient with painless progressive distal right leg weakness and calf enlargement. Electrodiagnostic studies and MAGNETIC RESONANCE IMAGING scanning were performed to evaluate the extent and cause of radicular damage as the etiology for unilateral calf hypertrophy. RESULTS: Examination and electrodiagnostic studies revealed right L5, right S1, and left L5 radiculopathies. Imaging studies demonstrated lumbar stenosis at L3-L4, L4-L5, and L5-S1 vertebral levels as well as L4-L5 and L5-S1 foraminal stenosis. After decompressive surgery the progressive nature of the patient's symptomatology halted, and he had partial resolution of his deficits. CONCLUSION: Although the patient had bilateral L5 radiculopathies, he only had hypertrophy in the distribution of his right S1 radiculopathy. This supports the hypothesis that dysfunction of the S1 nerve root or its distribution is a predisposing factor to develop neurogenic muscle hypertrophy. Furthermore, patients presenting with unilateral calf hypertrophy need a careful diagnostic evaluation for S1 radiculopathy as well as to exclude asymmetric presentation of systemic neuromuscular conditions. PMID- 12634578 TI - Supratentorial and infratentorial intraparenchymal hemorrhage secondary to intracranial CSF hypotension following spinal surgery. AB - STUDY DESIGN: A single case study was conducted. OBJECTIVES: To detail the occurrence of multiple intracranial, intraparenchymal hemorrhages following an iatrogenic acute reduction in cerebral spinal fluid pressure following excision of an intradural extramedullary spinal tumor. SUMMARY OF BACKGROUND DATA: Multiple supratentorial, intraparenchymal, intracranial hemorrhages following an acute reduction in cerebral spinal fluid pressure have not been previously reported in the literature. METHODS: A case report and literature review are presented. RESULTS: The patient made an uneventful full recovery. CONCLUSION: Intracranial hemorrhage must be considered in the differential diagnosis of patients presenting with persistent headache following spinal surgery when the dura has been breached and is associated with significant cerebrospinal fluid loss. PMID- 12634579 TI - Spinal cord injury following osteoporotic vertebral fracture: case report. AB - STUDY DESIGN: This report details the sudden catastrophic neurologic deterioration concerning a patient with an acute fracture of T5 associated with osteoporosis. OBJECTIVE: To describe a unique presentation of abrupt spinal cord injury in this condition. SUMMARY OF BACKGROUND DATA: Vertebral fractures occur frequently in osteoporosis, but associated spinal cord injury is a rare occurrence. When neurologic injury occurs it is generally of a very low incidence and with a gradual onset, and spontaneous recovery can be expected. Most reports in the literature are of delayed-onset paraplegia after vertebral collapse. We report a unique case of a sudden and catastrophic thoracic spinal cord injury occurring spontaneously. METHODS: An 88-year-old woman had a sudden thoracic vertebral fracture with complete spinal cord injury while attempting to get out of bed. She was treated with continuous oral corticosteroids for >40 years for both rheumatoid arthritis and restrictive lung disease. She was never investigated or treated for osteoporosis. RESULTS: Plain radiographs indicated multilevel osteoporotic fractures throughout the spine with diffuse, marked osteopenia. Magnetic resonance imaging scans showed changes in keeping with severe vertebral osteoporosis and spinal cord injury. She was treated nonoperatively in view of the extremely osteoporotic spine and her overall poor medical condition. Despite an initial improvement in her respiratory condition with medical therapy, she died 72 hours following admission. CONCLUSION: With increasing awareness of the morbidity attached to the osteoporotic spine, investigation and pharmacologic treatment are warranted and may reduce the possibility of catastrophic neurologic impairment as occurred in this case. PMID- 12634580 TI - Recurrent acute otitis media occurring within one month from completion of antibiotic therapy: relationship to the original pathogen. AB - OBJECTIVES: (1) To determine the relationship between acute otitis media (AOM) pathogens isolated in cases of early clinical recurrence of AOM (occurring within 1 month from completion of therapy) to the original pathogens causing the initial AOM episode; and (2) To determine whether shorter time intervals between completion of antibiotic therapy and clinical recurrences of AOM are associated with higher rates of true bacteriologic relapse. PATIENTS AND METHODS: From 1995 through 2000, 1077 infants and young children ages 3 to 36 months with AOM were enrolled in double tympanocentesis (performed on Day 1 in all patients and Days 4 to 6 in those initially culture-positive) studies. Of these, 834 (77%) completed successfully the antibiotic treatment [pathogen eradication on Days 4 to 6 of therapy or no pathogen on middle ear fluid (MEF) culture on Day 1 and clinical improvement at end of therapy]. Patients were followed for 3 to 4 weeks after completion of therapy, and additional MEF cultures were obtained if clinical recurrence occurred. True bacteriologic relapse was defined as the presence of a pathogen identical with that isolated before therapy by serotype and pulsed field gel electrophoresis for and by pulsed field gel electrophoresis for Streptococcus pneumoniae and beta-lactamase production for Haemophilus influenzae. RESULTS: MEF cultures were performed in 108 consecutive patients with early recurrent AOM. One hundred pathogens were isolated at recurrence in 88 of 108 (81%) patients: 54 H. influenzae; 45 S. pneumoniae; and 1 Moraxella catarrhalis. Most recurrent AOM episodes developed during the first 2 weeks of follow-up; 39 (36%), 38 (35%), 21 (19%) and 10 (9%) recurrent AOM episodes occurred on Days 1 to 7, 8 to 14, 15 to 21 and 22 to 28 after completion of therapy, respectively. In most patients these episodes were caused by a new pathogen. True bacteriologic relapses were found in 30 (28%) of 108 patients whose MEF cultures were positive for 35 pathogens: 13 of 108 (12%) S. pneumoniae; 12 of 108 (11%) H. influenzae; and 5 of 108 (5%) both. When timing of recurrent AOM after completion of therapy was analyzed, true bacteriologic relapses were found in 16 of 39 (41%), 10 of 38 (26%), 3 of 21 (14%) and 1 of 10 (10%) of all episodes on Days 1 to 7, 8 to 14, 15 to 21 and 22 to 28 after completion of therapy, respectively (P = 0.01). The respective rates for were 11 of 17 (65%), 3 of 10 (30%), 3 of 13 (23%) and 1 of 5 (20%) (P = 0.02). For H. influenzae the respective rates were 8 of 19 (42%), 9 of 23 (39%), 0 of 8 (0%) and 0 of 4 (0%) (P = 0.02). CONCLUSIONS: Most recurrent AOM episodes occurring within 1 month from completion of antibiotic therapy are in fact new infections. Most of the true bacteriologic AOM relapses occur within 14 days after completion of therapy, but even during this time interval most of the recurrences are caused by new pathogens. H. influenzae is very unlikely to cause true bacteriologic AOM relapses 14 days or later after completion of therapy. PMID- 12634581 TI - Forty-eight-week evaluation of lopinavir/ritonavir, a new protease inhibitor, in human immunodeficiency virus-infected children. AB - BACKGROUND: Lopinavir/ritonavir has demonstrated antiviral activity in the HIV infected adult. SUBJECTS: The objective of this study was to investigate a liquid coformulation of lopinavir/ritonavir, in combination with reverse transcriptase inhibitors, in HIV-infected children. METHODS: One hundred antiretroviral (ARV) naive and ARV-experienced, nonnucleoside reverse transcriptase inhibitor-naive children between 6 months and 12 years of age participated in this Phase I/II, open label, multicenter trial. Subjects initially received either 230/57.5 mg/m(2) or 300/75 mg/m(2) lopinavir/ritonavir twice daily; ARV-naive subjects also received stavudine and lamivudine, whereas ARV-experienced subjects also received nevirapine and one or two nucleoside reverse transcriptase inhibitors. Lopinavir/ritonavir pharmacokinetics, safety and efficacy were evaluated. RESULTS: All subjects were escalated to the 300/75 mg/m(2) twice daily dose based on results from an interim pharmacokinetic and safety evaluation. The pharmacokinetics of lopinavir did not appear to be dependent on age when dosing was based on body surface area but were decreased on coadministration with nevirapine. Overall 79% of subjects had HIV RNA levels <400 copies/ml at Week 48 (intent-to-treat: missing = failure). Mean increases in absolute and relative (percent) CD4 counts from baseline to Week 48 were observed in both ARV-naive subjects (404 cells/mm(3); 10.3%) and ARV-experienced subjects (284 cells/mm(3); 5.9%). Only one subject prematurely discontinued the study because of a study drug-related adverse event. CONCLUSIONS: The liquid coformulation of lopinavir/ritonavir demonstrated durable antiviral activity and was safe and well tolerated after 48 weeks of treatment in HIV-infected children. PMID- 12634582 TI - Lamivudine treatment for chronic hepatitis B infection in children unresponsive to interferon. AB - BACKGROUND/AIMS: Lamivudine is a potent inhibitor of hepatitis B virus (HBV) replication. This prospective open study reports the results of lamivudine treatment in children with chronic hepatitis B infection who did not respond to previous interferon treatment. PATIENTS AND METHODS: Lamivudine, 3 mg/kg/day (maximum, 100 mg/day), was given for 52 weeks to 20 children and adolescents, ages 8.5 to 19 years, with chronic hepatitis B infection who had been treated with interferon 2 to 5 years earlier. We evaluated virologic and biochemical responses, the occurrence of YMDD mutants and adverse effects. RESULTS: All children were HBV DNA+, hepatitis B e antigen (HBeAg) /anti-hepatitis B e antibody- at start of treatment. At the end of 1 year, HBV DNA declined by 95% in all patients, and 8 of 18 (44%) had sustained undetectable HBV DNA by hybridization assay. Median pretreatment alanine aminotransferase (ALT) x1.5 upper limit of normal decreased to ALT x0.9 upper limit of normal after 1 year. One child became HBeAg-negative. YMDD mutants were detected in 11 of 17 (65%) children after 1 year of lamivudine treatment. Among children with YMDD mutant variants, 54% maintained normal ALT values and 45% had undetectable HBV DNA by hybridization assay. No adverse effects were observed. CONCLUSIONS: Children with chronic hepatitis B infection treated with lamivudine after failure of interferon therapy had decreased HBV replication and improved ALT values. However, lamivudine treatment resulted in an exceptionally high rate of lamivudine resistant mutants and low HBeAg seroconversion rate. PMID- 12634583 TI - Homeopathy for childhood diarrhea: combined results and metaanalysis from three randomized, controlled clinical trials. AB - BACKGROUND: Previous studies have shown a positive treatment effect of individualized homeopathic treatment for acute childhood diarrhea, but sample sizes were small and results were just at or near the level of statistical significance. Because all three studies followed the same basic study design, the combined data from these three studies were analyzed to obtain greater statistical power. METHODS: Three double blind clinical trials of diarrhea in 242 children ages 6 months to 5 years were analyzed as 1 group. Children were randomized to receive either an individualized homeopathic medicine or placebo to be taken as a single dose after each unformed stool for 5 days. Parents recorded daily stools on diary cards, and health workers made home visits daily to monitor children. The duration of diarrhea was defined as the time until there were less than 3 unformed stools per day for 2 consecutive days. A metaanalysis of the effect-size difference of the three studies was also conducted. RESULTS: Combined analysis shows a duration of diarrhea of 3.3 days in the homeopathy group compared with 4.1 in the placebo group (P = 0.008). The metaanalysis shows a consistent effect-size difference of approximately 0.66 day (P = 0.008). CONCLUSIONS: The results from these studies confirm that individualized homeopathic treatment decreases the duration of acute childhood diarrhea and suggest that larger sample sizes be used in future homeopathic research to ensure adequate statistical power. Homeopathy should be considered for use as an adjunct to oral rehydration for this illness. PMID- 12634584 TI - Intussusception-associated hospitalization among Venezuelan infants during 1998 through 2001: anticipating rotavirus vaccines. AB - BACKGROUND: The first licensed rotavirus vaccine was withdrawn from use in the United States because of a low risk of intussusception. Consequently tests of new rotavirus vaccines will require some baseline knowledge of the rates and treatment of intussusception in countries where these vaccines will be tested. Therefore the objective of this study was to assess hospitalization rates and describe the epidemiologic and clinical characteristics of intussusception in Carabobo, Venezuela. METHODS: This study reviewed hospital data and clinical records of pediatric patients with intussusception admitted to eight hospitals in Carabobo between January 1, 1998 and December 31, 2001. RESULTS: For the 4-year period the average annual hospitalization rate for intussusception among infants (<1 year old) in Carabobo was 35 per 100 000 infants per year (range, 22 to 44), and intussusception was more common among boys (58 per 100 000 infants per year) than girls (29 per 100 000 infants per year) (P = 0.006). One of 67 cases occurred in the first 3 months of life (0 to 2), 55% (37) patients were 3 to 5 months old, 31% (21) were 6 to 11 months old and 12% (8) were > or =12 months old. Surgical reduction was required in 88% (52 of 59) of infants but only 1 required bowel resection. The monthly distribution of intussusception-associated hospitalizations showed no consistent seasonal pattern with the seasonality observed for patients hospitalized with rotavirus disease. CONCLUSIONS: This study provides some data about the incidence of intussusception and its epidemiology in a developing country where current or future field trials with rotavirus vaccine will be conducted. PMID- 12634585 TI - Evaluation of multiple drug therapy in human immunodeficiency virus-infected pediatric patients. AB - BACKGROUND: An aggressive therapeutic approach for treatment of HIV in adults consists of combining five or more concurrent antiretrovirals. The clinical benefits of this regimen are often accompanied by increased toxicities. We report the safety and tolerance of multiple drug therapy in HIV-infected children. METHODS: A retrospective chart review was performed to identify HIV-infected children who received > or =5 concurrent antiretrovirals or 4 antiretrovirals plus hydroxyurea. Treatment success was defined as > or =1 log(10) decrease in plasma HIV RNA from baseline any time during multiple drug therapy. Toxicities were defined as a >Grade 2 change from baseline in laboratory values. RESULTS: Twelve patients received multiple drug therapy for 6 months, and 42% of patients continued to receive therapy for at least 1 year. No Grade 3 or 4 toxicities or laboratory abnormalities were reported. Treatment success occurred in 8 (83%) of 12 patients. Adherence was a determining factor in treatment success or failure. CONCLUSIONS: Treatment of HIV-infected children with multiple drug therapy was well-tolerated in this cohort. Treatment success occurred in most patients, with adherence affecting patients' likelihood of success. Larger controlled clinical trials in this patient population are necessary to determine whether the benefit of this therapeutic approach outweighs potential risks. PMID- 12634586 TI - Epidemiology of neonatal infections: experience during and after hospitalization. AB - OBJECTIVE: We assessed the epidemiology of neonatal infections coming to medical attention among inpatient and outpatient newborn infants within a defined health care maintenance organization (HMO) population. DESIGN AND METHODS: This was a retrospective cohort study, using automated data from a large health maintenance organization and the hospital where the majority of HMO patients delivered. All infants delivered between October 1, 1990 and March 31, 1998 at the study hospital and receiving postdischarge care at the study HMO for the first 30 days of life were included. The outcomes assessed were 10 neonatal infection syndromes defined according to modified National Nosocomial Infection System criteria. RESULTS: There were 13 224 infants in the study cohort. Of these, 559 infants (4.2%) had 574 infections. Nonpneumonia respiratory infections were most common, accounting for 43% of all infections. Infections diagnosed in the outpatient setting comprised 63% of all infections. More serious infections (bloodstream infection, clinical sepsis and pneumonia) were typically diagnosed in the first few days of life and before nursery discharge. Infants with an infection had a significantly longer length of nursery stay (4.7 excess days among full term infants, P < 0.001), more office visits (397.3 vs. 273.3 per 100, P < 0.001) and more hospital admissions (16.7 vs. 3.6 per 100, P = 0.001) than uninfected infants. CONCLUSION: Infections were common during the first 30 days of life. The majority were noninvasive, were diagnosed in the outpatient setting and were most likely community-acquired. Serious infections were rare, occurred early in the neonatal period and were usually diagnosed before nursery discharge. PMID- 12634587 TI - Safety and therapeutic efficacy of artesunate suppositories for treatment of malaria in children in Papua New Guinea. AB - BACKGROUND: Although suppositories of artemisinin derivatives may be a valuable option for treatment of malaria in children when circumstances prevent oral and parenteral therapy, few confirmatory data have been published. METHODS: We assessed the safety and efficacy of rectal artesunate in 47 children ages 5 to 10 years with uncomplicated malaria acquired in a hyperendemic area of Papua New Guinea. Thirty were symptomatic and had Plasmodium falciparum parasitemia >2000/microl (Group 1), 12 had and either a parasitemia <2000/microl or minimal/no symptoms (Group 2) and 5 had Plasmodium vivax (Group 3). Each child received rectal artesunate 10 to 15 mg/kg at 0 and 12 h. After monitoring for 24 h, chloroquine plus sulfadoxine/pyrimethamine was given, and the patient discharged. RESULTS: Artesunate suppositories were well-tolerated. After 24 h only one child (from Group 1) had persistent parasitemia, and only one (from Group 3) had not defervesced. These two children received intramuscular quinine and recovered uneventfully. Three Group 2 children redeveloped fever and tachycardia at 24 h, but each responded to simple supportive measures and remained aparasitemic. CONCLUSIONS: Intrarectal artesunate is safe, effective initial treatment for uncomplicated malaria in children. A transient fever spike can sometimes occur after parasite clearance. We recommend that children with uncomplicated malaria receive two doses of > or =10 mg/kg rectal artesunate within the first 24 h. PMID- 12634588 TI - Mucosal immune response to specific outer membrane proteins of Moraxella catarrhalis in young children. AB - BACKGROUND: Moraxella catarrhalis is an important cause of otitis media. A number of candidate antigens for a future infant otitis media vaccine have been identified, but their mucosal immunogenicity induced by nasopharyngeal M. catarrhalis colonization has not been characterized. The aim of this study was to determine the salivary IgA response to M. catarrhalis outer membrane proteins (OMP) in young children. METHODS: Children ages 1 to 24 months evaluated for acute respiratory tract infection were prospectively enrolled. M. catarrhalis nasopharyngeal colonization was determined by (1) selective culture and (2) detection by reverse transcription-PCR of messenger RNA specific for the OMP UspA1 and UspA2. Salivary IgA responses were detected by immunoblot analysis of M. catarrhalis OMP. Isogenic knockout mutants for UspA1, UspA2, hemagglutinin (Hag), transferrin-binding protein B (TbpB) and CopB were constructed for identification of specific target OMP. RESULTS: Sixty-six patients were studied. The rates of M. catarrhalis colonization by culture, reverse transcription-PCR for messenger RNA and mRNA were 40, 94 and 58%, respectively. Anti-M. catarrhalis salivary IgA was detected in 62 patients (94%). IgA directed against a >250-kDa antigen (assigned to UspA1/UspA2 by mutant analysis) and a 200-kDa antigen (Hag) were detected in 65 and 70% of patients, respectively. Bands at 80 to 85 kDa (82%) consisted of IgA directed against monomeric UspA2, TbpB and CopB. CONCLUSIONS: colonization occurring in early infancy is associated with a consistent mucosal immune response directed against the UspA proteins, Hag and other OMP. The data suggest that several M. catarrhalis OMP are immunogens of the nasopharyngeal mucosal immune system of infants. PMID- 12634590 TI - One week versus four weeks of albendazole therapy for neurocysticercosis in children: a randomized, placebo-controlled double blind trial. AB - BACKGROUND: The appropriate duration of albendazole therapy in neurocysticercosis remains to be determined. Observations in small, uncontrolled clinical trials that short course therapy is as effective as longer regimens must be tested by proper trials. OBJECTIVE: To compare efficacy of 1 and 4 weeks of albendazole therapy in children with neurocysticercosis. STUDY DESIGN AND SETTING: A randomized, placebo-controlled, double blind clinical trial at the Pediatric Neurocysticercosis Clinic of Advanced Pediatric Centre, a tertiary care teaching hospital. SUBJECTS: We observed 122 consecutive children with seizures and either a single small enhancing computerized tomographic lesion or up to 3 lesions on head computerized tomography (CT) examination. INTERVENTION: All children received albendazole (15 mg/kg/day) for 7 days followed by either albendazole or placebo for the following 21 days according to their random number allocation. CT scans were repeated at 1 and 3 months after completion of therapy. Codes opened at the completion of study revealed that 60 children had received albendazole for 28 days (Group A) and 62 received albendazole for 7 days (Group B). All children were followed for at least 2 years. RESULTS: Complete resolution of lesions was similar in the two therapy groups on the first (42% vs. 39%) and second (77% vs. 79%) follow-up CT. Reduction in total number and size was also similar. Also the proportion of lesions that calcified (5% vs. 10%) did not differ significantly. Seizure control at 2 years was similar in the 2 groups. CONCLUSIONS: In this clinical trial 1 week of albendazole therapy was as effective as 4 weeks of therapy in children with neurocysticercosis having one to three lesions. PMID- 12634589 TI - Differences in nasopharyngeal bacterial flora in children with nonsevere recurrent acute otitis media and chronic otitis media with effusion: implications for management. AB - BACKGROUND: The interactions between nasopharyngeal flora and the individual entities covered by the broad term otitis media have not been completely elucidated. We investigated in infants and children ages 6 months to 7 years with nonsevere recurrent acute otitis media (rAOM) or with chronic otitis media with effusion (cOME): (1) the nasopharyngeal carriage rate and bacterial density of respiratory pathogens and alpha-hemolytic streptococci in comparison with healthy children; (2) the resistance pattern of respiratory pathogens; and (3) the relationship between the type of nasopharyngeal colonization and long term outcome. METHODS: Nasopharyngeal cultures were obtained from 85 children with rAOM,113 children with cOME and 55 controls. A semiquantitative analysis was used in the reading of cultures. A 12-week follow-up without treatment was planned. RESULTS: The carrier rate of respiratory pathogens was significantly greater in cOME (70%) than in rAOM (45%) (P = 0.0006) or controls (31%) (P < 0.0001). Similarly colonization density was significantly greater in cOME than in rAOM. The carriage rate and the colonization density of alpha-hemolytic streptococci were significantly lower in rAOM than in cOME or controls. The incidence of resistant (R) strains was greater in rAOM (Streptococcus pneumoniae penicillin-R, 24%; macrolide-R, 64%; Haemophilus influenzae amoxicillin-R, 24%) compared with cOME (S. pneumoniae penicillin-R,18%; macrolide-R, 44%; H. influenzae amoxicillin R, 5%) or controls (S. pneumoniae penicillin-R, 8%; macrolide-R, 23%; H. influenzae amoxicillin-R, 10%). During the follow-up period persistence of OME and occurrence of AOM were greater among carriers of respiratory pathogens at baseline. CONCLUSIONS: There are substantial differences in nasopharyngeal flora between children with nonsevere rAOM and children with cOME. The results of nasopharyngeal cultures should be taken into account to avoid treatment with drugs that are ineffective and likely to select resistant organisms. PMID- 12634591 TI - Cold-adapted, live attenuated intranasal influenza virus vaccine. PMID- 12634592 TI - Traveling with children. PMID- 12634593 TI - Metastatic intracranial abscesses of bronchopulmonary origin. AB - Intracranialabscesses are serious, life-threatening infections despite recent advances in diagnosis and treatment. We report the case of a child with metastatic brain abscesses and a chronic, sharp foreign body of the bronchus. Its presentation and endoscopic appearance and the microbiology of the abscesses are detailed. The detection of bacteria of respiratory origin should lead to a search for a bronchopulmonary source of contamination. PMID- 12634594 TI - High fevers as a cause of central nervous system sequelae. PMID- 12634595 TI - Polymerase chain reaction diagnosis of human immunodeficiency virus infection. PMID- 12634596 TI - Varicella, herpes zoster and dissemination. PMID- 12634597 TI - Supplementary table: epidemiology of neonatal infections: experience during and after hospitalization. PMID- 12634598 TI - Haematological problems in pregnancy. AB - PURPOSE OF REVIEW: Haematological disorders of pregnancy are common reasons for referral. It is reasonable then to devote a review to recent publications on these issues. RECENT FINDINGS: Several narrative reviews on the management of venous thromboembolic disease in pregnancy provide exceptional guidance. They highlight, however, that most of what is done is still based on opinions from highly qualified people, extrapolated from the non-pregnant literature but not involving randomized trials on pregnant patients. It is apparent that a consensus is nearing on the management of both pregnancy-related thrombocytopenia and idiopathic (immune) thrombocytopenic purpura. Several randomized controlled trials are reported on the treatment of non-deficiency anaemia, using oral iron, intravenous iron and erythropoietin, which contribute to the debate on the appropriate and best intervention for this common problematic issue. SUMMARY: This review demonstrates the need for appropriately sized randomized trials on haematological issues in pregnancy. With heparin becoming the most common medication being administered prophylactically and therapeutically in pregnancy, it is important that quality trials are performed to guide its sensible utilization. PMID- 12634599 TI - Recent evidence associated with the condition of preterm prelabour rupture of the membranes. AB - PURPOSE OF REVIEW: The published literature on preterm prelabour rupture of the membranes is voluminous yet despite advances in obstetric and neonatal care, the problem remains a major cause of perinatal mortality and morbidity. The purpose of this review is to present recent evidence pertaining to the role of inflammatory mediators such as cytokines and the tissue damage and long-term handicap they cause, the molecular biology and physiology of membrane structure, the role of host susceptibility and the genetics of preterm birth and therapeutic options for the management of preterm prelabour rupture, including antibiotics, amnioinfusion and special situations. RECENT FINDINGS: Neonatal morbidity from preterm prelabour rupture of the membranes is mainly related to oligohydramnios and pulmonary hypoplasia. Occupational factors have a significant effect on the occurrence and outcome following rupture. Matrix metalloproteinases control growth and remodelling of the pregnant uterus, placenta and membranes and are linked to a genetic predisposition to preterm birth through gene expression and variation. Transvaginal ultrasound scan, oncofetal fibronectin and the presence of abnormal genital tract flora (bacterial vaginosis) in pregnancy may help in the prediction of preterm birth. SUMMARY: Preterm prelabour membrane rupture remains a management problem, particularly at very early gestations, yet obstetric and neonatal care can make a difference to outcome. While at early gestations the prognosis is poor, it is not hopeless. Careful selection of the recent literature on the subject might interest and inform those faced regularly with the problem, prevent therapeutic nihilism, promote confidence in our ability to make a difference and realise that we are not alone when faced with the therapeutic dilemma that is this condition. PMID- 12634600 TI - Pituitary and adrenal disorders complicating pregnancy. AB - PURPOSE OF REVIEW: Confidential enquiries of maternal deaths in the UK for 1997 1999 revealed that the number of indirect deaths from medical conditions exacerbated by pregnancy was greater than deaths from conditions directly caused by pregnancy. Indirect maternal deaths accounted for 36% of all deaths. Although neoplastic disorders of the pituitary and adrenal glands are very rare during pregnancy, they can be potentially fatal. The rarity of these disorders may pose difficulty in formulating an optimum plan of management. Biochemical tests that can be performed in the non-pregnant state may be of little value during pregnancy. The presence of a fetus may also influence the type and timing of intervention. RECENT FINDINGS: A few case reports have been reported in the literature, but unfortunately no large series to develop guidelines or protocols. However, endoscopic surgical procedures have recently been used in their management with good outcomes. New drugs such as octreotide and lanreotide are increasingly being used during pregnancy. Although too soon to assess the safety of these new treatments, results so far have been very encouraging. SUMMARY: Awareness of these potentially fatal neoplastic disorders of the pituitary and adrenal glands, coupled with early diagnosis and appropriate treatment, may help improve maternal and fetal outcomes. Advances in imaging techniques and endoscopic procedures are also likely to change clinical practice. Various new treatment options are being reported in the literature, but they are not supported by randomized, controlled trials. Further research is needed to determine their safety and efficacy during pregnancy. PMID- 12634601 TI - Psychological aspects of perinatal loss. AB - PURPOSE OF REVIEW: Parents are attached to their unborn children, and loss around the time of birth is a serious trauma. Parental grief is a normal response, and may last for many months. Clinicians have always sought to implement practices that will help recovery, and for a generation, have advocated that parents have contact with the body of their dead infant, believing that this will facilitate mourning. Review of the literature shows that no previous systematic evidence has ever been offered to support this practice. RECENT FINDINGS: Recent research throws doubt on the therapeutic value of the practice of encouraging contact with the dead infant. These findings are outlined and discussed. SUMMARY: Following an outline of what is already known about the psychological effects of perinatal loss, the paper focuses on the evidence that aspects of psychosocial management are associated with better outcomes. It goes on to describe the cultural context in which psychosocial management changed and parents were first encouraged to see and handle their dead infant, and explores the distinction between the medical and cultural models which may frame this behaviour in very different ways. Finally, it briefly discusses the ethical position of staff who advise parents about their choice to see or not see their infant's body. PMID- 12634602 TI - Outcome of triplets and high-order multiple pregnancies. AB - PURPOSE OF REVIEW: To present data related to the outcome of triplets and high order multiple pregnancies. RECENT FINDINGS: Current frequencies of high-order multiple pregnancies in most developed countries range between 400 and 800% above the rates observed in the late 1970s. Of particular importance is the striking increase in pregnancies in older mothers. These epidemiological trends are the result of modern infertility treatments. The improved outcome of triplets may be attributed to close antenatal and perinatal care, both of which are more likely to be implemented in patients who can afford treatment for infertility. The overall odds of delivering at least one triplet infant weighing less than 1000 g is approximately 10%. The neonatal complications among triplets weighing less than 1500 g at birth are not much different from those among twins or singletons, except for a higher neonatal mortality rate. Despite this, the frequency of cerebral palsy bears a significant exponential relationship to the number of fetuses, and iatrogenic multiple births are clearly implicated in the increased cerebral palsy rate. In the past few years the perinatal mortality rate for triplets has been approximately 110/1000, three-quarters of which represent neonatal deaths. SUMMARY: Available data imply that in order to improve outcomes further, all multiple pregnancies deserve optimal perinatal care, something that is not always obtainable. PMID- 12634603 TI - Amniotic fluid embolism. AB - PURPOSE OF REVIEW: To review the recent literature on amniotic fluid embolism and how it may influence the clinical management and further study of the condition. Morbidity and mortality from amniotic fluid embolism in the international context will be described, given the recent Confidential Enquiries into Maternal Deaths in the United Kingdom and other studies. With this rare condition we need to look for clues as to how to facilitate diagnosis and improve outcomes. RECENT FINDINGS: Amniotic fluid embolism continues to be a leading cause of maternal death. There has been a decrease in mortality from amniotic fluid embolism in the UK from 5.1 to 3.7 per million maternities, but it is still the fifth greatest cause of direct maternal death. In France, 13% of deaths are caused by amniotic fluid embolism, the third highest cause. In Singapore, a study of postmortems found that over 30% of direct maternal deaths were caused by amniotic fluid embolism, the most common cause. Case-specific mortality may not be as high as previously thought. The early data from the UK Register of cases show only 16% mortality, although there is significant maternal and neonatal morbidity. Early diagnosis may be the best way to improve outcomes. Case reports suggest that plasma exchange techniques may be helpful after initial resuscitation. SUMMARY: With a rare condition additions to the literature are sparse. Early consideration of the diagnosis after prompt resuscitation is needed. Further data are needed to advance beyond this. PMID- 12634604 TI - Management of previous cesarean section. AB - PURPOSE OF REVIEW: The management of cesarean sections causes much controversy among healthcare providers, patients, and insurers. A trial of vaginal birth after previous cesarean is reported to be a safe and practical method to reduce the rate of cesarean sections. The popularity of vaginal birth after previous cesarean has increased over the past two decades, but rates have recently started to decline again. This review will evaluate recent literature that might be responsible for this reversal in trend. RECENT FINDINGS: Earlier studies on previous cesarean section pregnancies focused primarily on the success rate of vaginal birth after previous cesarean, which is reported to be 60-80%. Recent large, retrospective, population-based cohort studies examined the maternal and neonatal safety of trial of labour compared with elective repeat cesarean delivery, and confirmed that the risks of uterine rupture and neonatal mortality were significantly increased after trial of labour, particularly when induced with prostaglandins. However, the absolute risk of adverse events remains small. The maternal and neonatal morbidity risk increases when vaginal birth after previous cesarean attempts fails, which emphasizes the importance of careful case selection. SUMMARY: Recent studies highlighted the risks of attempted vaginal birth after previous cesareans, especially when trials fail, but have not addressed the long-term risks of an elective repeat cesarean delivery. The assessment of treatment risks by observational studies is subject to bias, because the different treatment groups may not be comparable at the outset. In the absence of better data, the counselling of such women must currently be based on this evidence. PMID- 12634605 TI - First trimester anatomy scan: pushing the limits. What can we see now? AB - PURPOSE OF REVIEW: The aim of this review article is to familiarize the reader with the current developments dealing with ultrasound during the first trimester of pregnancy. RECENT FINDINGS: The main emphasis of this article is to review the anatomy seen during the first trimester and to bring to the reader the fact that the first anatomical survey of the fetus does not have to wait until the 18th to 20th week of gestation. The fetus can be surveyed adequately as early as the 12th week of gestation and ideally between the 13th and 14th weeks. Of course, not all malformations will be detected during the first trimester, and a follow-up scan between 20 and 23 weeks is indicated. SUMMARY: The ongoing improvements in ultrasound technology allow us to see more details of the embryo or fetus at earlier gestational ages than ever before. This is resulting in a rethinking of when the first anatomical survey should be performed. PMID- 12634606 TI - Cardiac anatomy screening: what is the best time for screening in pregnancy? AB - PURPOSE OF REVIEW: To consider the ideal gestational age for cardiac evaluation during pregnancy. RECENT FINDINGS: Screening the heart during routine obstetric ultrasound has become well established and is increasingly successful in the initial detection of major congenital heart disease. When the option of termination of pregnancy is available, the earlier the diagnosis of any major fetal malformation is made, the better for the patient. An important group who have recently been found to be at increased risk of fetal heart malformation are those with increased nuchal translucency measurements, who are identified between 11 and 14 weeks. Thus, screening for fetal anomalies is being attempted much earlier in pregnancy, aided by advances in technology. However, some forms of cardiac malformations do not become evident until the third trimester of pregnancy; as a result, some of the late-developing lesions may go undetected during very early evaluation. SUMMARY: The ideal timing for screening is a compromise between obtaining adequate images for diagnosis in the majority of routine patients, scanning sufficiently late not to miss late-developing lesions and yet offering diagnosis as early as possible for parents to consider their options, if there are any applicable to their particular diagnosis. For low-risk patients, the best compromise appears to be at around 20 weeks of gestation. For patients at increased risk of congenital heart disease, such as those found to have substantially increased nuchal translucency or those with a family history of the disease, an initial scan to exclude major malformations should be performed by the fetal cardiology expert at 12-14 weeks, with follow-up at around 20 weeks to exclude more minor defects and those lesions which may become evident later. PMID- 12634607 TI - Comprehensive assessment of fetal wellbeing: which Doppler tests should be performed? AB - PURPOSE OF REVIEW: Doppler applications in pregnancy are expanding exponentially. Flow velocity waveforms provide important information 12 weeks to term, from maternal vessels, placental circulation and fetal systemic vessels, with implications for both mother and fetus. As applications proliferate, awareness of the complexity of fetal and placental circulations, in normal pregnancy and in sequential responses to compromise, has also grown. The necessary data are now available to establish core values in Doppler evaluation for at-risk pregnancies. RECENT FINDINGS: Uterine arteries depict maternal vascular effects of the invading placenta, predicting the frequency and severity of pre-eclampsia and intrauterine growth restriction. New evidence suggests early treatment based on this principle, significantly reduces these impacts. Umbilical artery Doppler reflects downstream placental vascular resistance, strongly correlated with intrauterine growth restriction and the multisystem effects of placental deficiency. Abnormalities are progressive, with reduction, loss, and finally a reversal of diastolic flow. When umbilical arteries become abnormal, the differentiation of fetal status requires Doppler information from systemic vessels. Middle cerebral artery changes begin when the redistribution of cardiac output reflects rising placental resistance, demonstrating 'brain sparing' when cerebrovascular dilation occurs. In the compromised intrauterine growth retarded fetus, precordial veins illustrate fetal cardiac function, changing as the respiratory status declines. This Doppler information is combined with biophysical profile scoring to determine the need for and timing of intervention. SUMMARY: Doppler evaluation of at-risk pregnancies provides crucial prognostic and diagnostic detail about placentation and fetal adaptation. What has been research detail is now becoming the standard of care, in comprehensive fetal maternal assessment. PMID- 12634608 TI - Chorionic villus sampling and amniocentesis: what are the risks in current practice? AB - PURPOSE OF REVIEW: Public demand for genetic counselling and prenatal diagnosis has increased during the past decade. As invasive diagnostic methods, such as chorionic villus sampling and amniocentesis, still have an important role to play in evaluating the fetus, one of the most important questions to address during genetic counselling is the procedure-related risk of these techniques. RECENT FINDINGS: The possible factors modifying the specific risk of the actual fetus are discussed, together with factors that have an impact on procedure-related fetal loss and other complications. Risk factors regarding twin pregnancies, first and second-trimester chorionic villus sampling, early and mid-trimester amniocentesis are discussed separately. New developments have recently occurred in the laboratory techniques used in prenatal diagnosis. Their impact on genetic counselling and the employment of invasive techniques are also addressed. SUMMARY: During genetic counselling, an individually tailored risk assessment needs to be established before any invasive procedure. This should take into account all the factors modifying the specific risk for aneuploidy or other disorders of the fetus, as well as the actual procedure-related risks. PMID- 12634609 TI - Invasive procedures in multiple gestations. AB - PURPOSE OF REVIEW: The incidence of multiple gestations has increased significantly with advances in assisted reproductive techniques resulting in a concomitant increase in invasive procedures in these pregnancies. Commonly performed invasive procedures include amniocentesis, chorionic villus sampling, multifetal pregnancy reduction, and selective termination. Amniocentesis and chorionic villus sampling are also performed in singleton pregnancies, while multifetal pregnancy reduction and selective termination are procedures that are unique to multiple gestations. RECENT FINDINGS: With increased operator experience, pregnancy loss rates after chorionic villus sampling, multifetal pregnancy reduction, and selective termination have decreased to acceptably low levels. Amniocentesis and chorionic villus sampling continue to have similar loss rates in experienced hands. A recent study suggests that amniocentesis in twins may have a higher post-procedural loss rate than in singletons; this may be due to the higher background loss rate of twins. There has been a recent increase in multifetal pregnancy reduction to a singleton with a trend towards improved outcomes over reduction to twins; future studies should focus on whether this confers a definitive advantage. Newer data suggests that selective termination after 20 weeks gestation in experienced hands does not increase loss rates over those procedures performed before 20 weeks. Newer techniques, such as cord coagulation, continue to be developed for selective termination in monochorionic pregnancies, though still with considerable morbidity and mortality. SUMMARY: In summary, invasive procedures in multiple gestations are now commonly performed with. It is our hope that primary prevention of high order multiple pregnancies by optimization of assisted reproductive techniques will decrease the need for these procedures. PMID- 12634610 TI - Monochorionic diamniotic twins: complications and management options. AB - PURPOSE OF REVIEW: Monochorionic compared with dichorionic twins have disproportionately high fetal loss rates, perinatal mortality and morbidity. This is because of the unpredictable vascular anastomoses and the often asymmetrical distribution of the single placenta between both twins. RECENT FINDINGS: The pathophysiology of twin-to-twin transfusion syndrome is usually explained on an angioarchitectural basis, although certain hemodynamic and hormonal factors also may be involved. The results of the large randomized trials on amnioreduction, fetoscopic laser coagulation and septostomy are still awaited. An update is given on hardware and instruments required for fetoscopic laser. Subsequently, the problem of the monochorionic twin set with severe early discordant growth is addressed. Several etiological mechanisms have been proposed, but little is known of its natural history. Also, umbilical artery Doppler waveforms may not have the same predictive value as in singletons. Prophylactic laser coagulation of the vascular anastomoses to protect against the adverse effects of single intrauterine demise, has so far not been shown to confer any benefit in outcome. Finally, pathophysiology and management of discordant structural and chromosomal anomalies in monochorionic twins are discussed. Laser and monopolar coagulation, which can be introduced through a needle, may be used for selective feticide in early pregnancy or low hemodynamic conditions. Bipolar coagulation seems more effective at later gestational ages and normal hemodynamic conditions. SUMMARY: Our insight into the complications associated with monochorionic twins has increased in recent years. It is hoped that this will lead to better surveillance and ultimately an improved outcome for these high-risk pregnancies. PMID- 12634611 TI - Fetal cell microchimerism: helpful or harmful to the parous woman? AB - PURPOSE OF REVIEW: Fetal cells enter the maternal circulation during most pregnancies and can persist in maternal blood and tissues after delivery. Concerns with regard to the histocompatibility of these fetal cells have raised the question of the long-term consequences of an immune response on maternal health. In the past few years, many investigators have demonstrated an association between the persistence of fetal cells in maternal tissues and blood and maternal autoimmune disease, especially systemic sclerosis. In this review we will summarize more recent data that provide a new insight into bi-directional feto-maternal cell trafficking. RECENT FINDINGS: Persisting fetal cells have been found in the tissue of women affected with endocrine or infectious disease as well as healthy parous women. SUMMARY: These data suggest the possibility that fetal microchimeric cells may also participate in the maternal physiological response to tissue injury. The medical consequences of pregnancy, therefore, appear to extend well beyond delivery. PMID- 12634612 TI - Toxicity and disposition of TLC ELL-12 (liposomal antitumor ether lipid) in Sprague-Dawley rats. AB - TLC ELL-12 is a liposomal formulation of the antineoplastic L-O-octadecyl-2-O methyl-sn-glycero-3-phosphocholine [L-ET-18-OCH3 (EL)]. The purpose of these studies was to characterize the toxicity and disposition of [N-methyl-14C] L-ET 18-OCH3 administered as TLC ELL-12. Rats received TLC ELL-12 by i.v. infusion into a tail vein as a single 12.5 or 62.5 mg/kg dose or as five daily doses at 12.5 mg/kg (cumulative dose of 62.5 mg/kg). Whole blood and tissue samples were collected over 24 h, and assayed for total and EL-specific radioactivity. The amounts of radioactivity in urine, bile, injection site and carcass were determined for up to 48 h. TLC ELL-12 was well tolerated in male and female rats in single doses up to 37.5 mg/kg. The minimum lethal dose was 112.5 mg/kg. Doses of 12.5 mg/kg (no observed adverse effects) and 62.5 mg/kg (approximate maximum tolerated dose) were chosen for further study. The pharmacokinetics of EL given as TLC ELL-12 were non-linear with a disproportionate increase in AUC at the higher dose. Daily dosing at 12.5 mg/kg did not result in accumulation in the blood. The highest concentrations of EL at 24 h after dosing were in the spleen and liver. Virtually no radioactivity was recovered in the urine or bile of rats, most remaining in the carcass and injection site (tail). After a 12.5 mg/kg dose of TLC ELL-12, the levels of EL in the blood and most tissues examined were well above the levels that inhibit tumor growth and may therefore be therapeutically active. PMID- 12634613 TI - Interaction of 5-aza-2'-deoxycytidine and depsipeptide on antineoplastic activity and activation of 14-3-3sigma, E-cadherin and tissue inhibitor of metalloproteinase 3 expression in human breast carcinoma cells. AB - Genes that suppress tumorigenesis can be silenced by epigenetic events, such as aberrant DNA methylation and modification of chromatin structure. Inhibitors of DNA methylase and histone deacetylase (HDAC) can potentially reverse these events. The aim of this study was to determine the in vitro antineoplastic activity of 5-aza-2'-deoxycytidine (5-AZA-CdR), a potent inhibitor of DNA methylase, in combination with depsipeptide (depsi), an inhibitor of HDAC, on human breast carcinoma cells. We observed a synergistic antineoplastic interaction between 5-AZA-CdR and depsi in their capacity to inhibit colony formation of Hs578T and MCF-7 breast carcinoma cells. In order to understand the molecular mechanism of this interaction, we investigated the effect of these drugs on the activation of the 14-3-3sigma, E-cadherin and tissue inhibitor of metalloproteinase 3 (TIMP3) cancer-related genes, which were reported to be silenced by aberrant methylation in many breast tumor cell lines. 14-3-3sigma was reported to produce G cell cycle arrest following DNA damage. E-cadherin and TIMP3 function as suppressors of tumor metastasis. Semi-quantitative RT-PCR was used to determine the effect of the co-administration of 5-AZA-CdR and depsi on four breast carcinoma cell lines for the reactivation of these genes. We observed a synergistic activation of E-cadherin by the combination in Hs578T, MDA-MB-231 and MDA-MB-435 tumor cells. For 14-3-3sigma, we demonstrated an additive to synergistic activation by the combination for Hs578T and MDA-MB-435 tumor cells, respectively. In the MCF-7 tumor cells, the drug combination produced a synergistic activation of TIMP3. The association between the synergistic antineoplastic activity and the synergistic activation of the target genes in this study suggests that the mechanism of anticancer activity of 5-AZA-CdR, in combination with depsi, is probably related to their enhanced activation of different types of tumor suppressor genes that have been silenced by epigenetic events.(2) PMID- 12634614 TI - Metabolism of paclitaxel in mice. AB - Previous mass balance studies in humans and mice have shown that the fecal and urinary recovery of paclitaxel and known metabolites (3' -hydroxypaclitaxel, 6alpha-hydroxypaclitaxel and 3',6alpha-dihydroxypaclitaxel) was not complete. Obviously this discrepancy is caused by the existence of other yet unknown metabolites. Mdr1a/1b(-/-) mice excrete very low quantities of unchanged paclitaxel. We have therefore used these mice receiving i.v. [3H]paclitaxel to further study the metabolic fate of paclitaxel. The major part of the radiolabel, being 70%, was excreted in the feces. A lipophilic sample, containing about 70% of the radioactivity present in the feces sample, was obtained by diethyl ether extraction. The aqueous residue containing about 30% of the radioactivity was further extracted using methanol. The high-performance liquid chromatography (HPLC) chromatograms of the lipophilic and aqueous sample revealed two and five putative new metabolites of paclitaxel, respectively. The HPLC fractions containing substantial amounts of radioactivity were subjected to tandem mass spectrometry. Two novel monohydroxylated paclitaxel structures were identified, which are probably 2m-hydroxypaclitaxel and 19-hydroxypaclitaxel, structures previously identified in rats. Including these metabolites, about 60% of the mass balance of paclitaxel could be quantified. PMID- 12634615 TI - Magnolol induces apoptosis in human leukemia cells via cytochrome c release and caspase activation. AB - Magnolol, isolated from the stem bark of Magnolia officnalis, was found to inhibit proliferation of human HL-60 cells and Jurkat T leukemia cells via inducing apoptosis in a dose- and time-dependent manner. By contrast, magnolol did not cause apoptosis in neutrophils and peripheral blood mononuclear cells of healthy donors. Apoptosis was determined by detection of DNA fragmentation in gel electrophoresis, morphological alternations by flow cytometry, quantification of phosphatidylserine externalization by Annexin V labeling and oligonucleosomal DNA content by TUNEL labeling. Activation of caspase-9, -3 and -2, and the proteolytic cleavage of poly(ADP-ribose) polymerase were found during apoptosis induced by magnolol. In addition, both pan-caspase and selective caspase-9 inhibitor blocked magnolol-induced apoptosis. The apoptosis could also be partially attenuated by caspase-3 and -2 inhibitors. Magnolol induced the reduction of mitochondrial transmembrane potential and the release of cytochrome c into cytoplasm. In conclusion, our findings indicate that magnolol-induced apoptotic signaling is carried out through mitochondria alternations to caspase-9 and that then the downstream effector caspases are activated sequentially. Magnolol could be a potentially effective drug for leukemia with low toxicity to normal blood cells and it merits further investigation. PMID- 12634616 TI - Synthetic chenodeoxycholic acid derivative HS-1200-induced apoptosis of p815 mastocytoma cells is augmented by co-treatment with lactacystin. AB - The antitumor activity of a synthetic chenodeoxycholic acid derivative, HS-1200, on the p815 mastocytoma cell line was investigated. We present several lines of evidence indicating that HS-1200 at 35 microM induced apoptosis of p815 cells. Reduction of mitochondrial membrane potential, the release of cytochrome to cytosol, activation of caspase-3, nuclear condensation, production of poly(ADP ribose) polymerase cleavage, generation of DNA fragmentation and nuclear condensation were demonstrated. Importantly, HS-1200 inhibited proteasome activity. Next, the combination treatment of HS-1200 or a proteasome inhibitor lactacystin was undertaken. Although the single treatment of 20 microM HS-1200 or 1 microM lactacystin induced apoptosis slightly, the combination treatment of them augmented prominently the extent of apoptosis. The combination therapy of HS 1200 and lactacystin could be potentially a therapeutic strategy reducing the extent and severity of treatment-related toxicity. PMID- 12634617 TI - Red blood cells: a neglected compartment in topotecan pharmacokinetic analysis. AB - Previously, a gender dependency of topotecan was found in the pharmacokinetics in the plasma compartment. Here, we prospectively studied the red blood cell (RBC) partitioning of topotecan and evaluated its consequences for overall drug disposition. Blood samples were obtained from 12 patients receiving cisplatin followed by i.v. topotecan. Topotecan pharmacokinetic analysis was performed in whole blood, plasma and RBCs. Significantly slower clearance was noted in females (n=7) compared to males (n=5) for lactone and total topotecan in plasma (p<0.0001), and for total drug in RBCs (p=0.027), but not in whole blood. In addition, no gender-dependent differences were observed in the terminal half lives of topotecan in any of the compartments. The area under the curve ratios for RBC total to plasma lactone were 2.53+/-0.0640 and 2.13+/-0.442 in males and females, respectively. Hence, topotecan displays preferential affinity for RBCs compared to plasma, although these cells do not act as a depot in which drug accumulates over time. RBCs thus play a principal role in the distribution kinetics of topotecan and have a major impact on its plasma pharmacokinetics. The data warrant a change from current practice in pharmacokinetic studies with this agent and provide further evidence that, in general, the choice of the appropriate assay matrix should be rationally based. PMID- 12634618 TI - Weekly docetaxel in patients with pretreated metastatic breast cancer: a phase II trial. AB - Docetaxel has consistently demonstrated its high activity as an antineoplastic agent in the treatment of metastatic breast cancer. However, 90% of patients receiving the recommended dose of 100 mg/m2 every 3 weeks will develop grade 3 or 4 neutropenia. Recent data suggest that the safety profile of a weekly docetaxel regimen compared favorably with the standard 3-week schedule. Thus, we initiated a phase II study to assess the efficacy and toxicity of weekly docetaxel in pretreated patients with metastatic breast cancer. Twenty patients with advanced, anthra-cycline-refractory breast cancer were included in this phase II trial. Docetaxel was administered at a starting dose of 40 mg/m2, repeated once a week for 3 consecutive weeks followed by a 1-week rest period (1 cycle). Patients were evaluated for tumor response every 8 weeks (after every other cycle). Therapy was continued for a maximum of six courses in patients showing tumor response or stable disease. Twenty patients received a total of 204 weekly infusions of docetaxel. The mean number of treatments was 10.2 (range 1-18). Eighteen patients were assessable for response. Five patients achieved a partial response and six patients showed either stable disease or a minor response. Seven patients had disease progression. The median survival was 7.8 months. Grade 3/4 leukopenia occurred in two patients. No other grade 3 or 4 hematologic toxicities were observed. The following grade 3/4 non-hematologic toxicities were seen: nausea/vomiting (one patient), infection (one patient), mucositis (two patients) and diarrhea (one patient). Three patients withdrew from the study due to dose limiting toxicities (one due to severe neutropenia and two due to mucositis). We conclude that administration of docetaxel at a dose of 40 mg/m2 was effective and well tolerated even in heavily pretreated patients with metastatic breast cancer. This regimen is associated with only mild myelosuppression. PMID- 12634619 TI - Pharmacokinetics of doxorubicin administered i.v. as Myocet (TLC D-99; liposome encapsulated doxorubicin citrate) compared with conventional doxorubicin when given in combination with cyclophosphamide in patients with metastatic breast cancer. AB - Myocet (TLC D-99) is a liposomal formulation of the anti-neoplastic drug doxorubicin with an improved therapeutic index compared with conventional doxorubicin. The objective of this study was to assess the plasma disposition of doxorubicin when administered i.v. as TLC D-99 and to compare this to conventional drug. Metabolite (doxorubicinol) plasma levels were also quantitated in both treatment groups. Plasma was collected during the first course of treatment from 10 patients receiving TLC D-99 60 mg/m and 10 receiving conventional doxorubicin 60 mg/m2, each with cyclophosphamide 600 mg/m2. Samples were assayed for total doxorubicin (all doxorubicin regardless of whether it is encapsulated or not), encapsulated doxorubicin (TLC D-99 group only) and doxorubicinol using high-performance liquid chromatography. Plasma concentrations of total doxorubicin were higher in patients receiving TLC D-99 than in patients receiving conventional doxorubicin. The clearance of total doxorubicin after administration of TLC D-99 was lower (approximately 9-fold) and the volume of distribution at steady state was less (25-fold) than that of doxorubicin after conventional drug. Doxorubicinol was detected in the plasma of all patients in both treatment groups. The mean AUC(0-infinity) of doxorubicinol for patients receiving TLC D-99 (1.5+/-0.4 M x h) was not statistically different than that in patients receiving conventional doxorubicin (1.8+/-0.4 M x h), although the appearance of the peak doxorubicinol concentration occurred later and was lower in patients receiving TLC D-99. There was a correlation between the plasma AUC(0 infinity) of total doxorubicin and the degree of myelosuppression in patients receiving conventional doxorubicin, but this correlation was not found in patients receiving TLC D-99. PMID- 12634620 TI - Doxil-induced regression of pleuro-pulmonary metastases in a patient with malignant meningioma. AB - Metastatic meningioma is a rare disease, which has no effective chemotherapy. We report on a treatment of this condition with Doxil, a liposomal doxorubicin formulation. A 60-year-old woman with massive pleuro-pulmonary metastases from recurrent cranial meningioma was treated with Doxil (50-37.5 mg/m2) for 18 months with near-complete resolution of metastases and disappearance of pleural fluid. The only significant toxicities observed were stomatitis and hand-foot syndrome, which resolved with dose reduction and increase of dosing intervals. Doxil was cleared very slowly in this patient with a monoexponential half-life of 108 h. The patient remains in near-complete response for 6 months after treatment discontinuation. This is the first report on an effective chemotherapy in a patient with typical metastatic meningioma. The exact mechanism accounting for such an effective drug action is not clear, but may be related to a particularly high microvascular permeability to the liposome carriers in these metastatic lesions. PMID- 12634621 TI - Merkel cell carcinoma developing after antithymocyte globulin and cyclosporine therapy for aplastic anemia. AB - We report a patient who developed Merkel cell carcinoma (MCC) after treatment with antithymocyte globulin and cyclosporine for aplastic anemia. The clinical course was progressive and poor prognosis. Although MCC is relatively rare in second cancers arising after immunosuppressive therapy, patients should be closely monitored for the development of this complication as well as other second malignancies. PMID- 12634622 TI - Shedding a new light on Bethesda III: new technologies in cervical screening. PMID- 12634623 TI - Simplification of the new Bethesda 2001 classification system. AB - In 1989, a National Cancer Institute workshop resulted in the development of the Bethesda System for cytologic reporting of Papanicolaou smears. In the Bethesda III System (2001), potentially premalignant squamous lesions fall into 3 categories: Atypical squamous cells (ASC), low-grade squamous intraepithelial lesions (LGSIL), and high-grade intraepithelial lesions (HGSIL). The ASC category is subdivided into 2 categories: Those of unknown significance (ASC-US) and those in which high-grade lesions must be excluded (ASC-H). Further revision included the elimination of the category "benign cellular changes," which is now referred to as "negative for intraepithelial lesion or malignancy." The revision also refined the criteria for abnormal glandular epithelium, including atypical glandular cells (AGC). The principal modification in the updated system is the revision of the ASC category, which was done to facilitate triage of women for more intense screening when a true suspicion of a high-grade lesion was present. PMID- 12634625 TI - The science behind the effectiveness of in vivo screening. AB - Meta-analyses of studies examining the sensitivity of the Papanicolaou (Pap) smear have revealed that up to 50% evade detection by a single Pap test. Liquid based preparation techniques have improved the false-negative rate; however, an apparent irreducible 20% false-negative rate still exists. Emerging data now show that aberrant expression of adhesion molecules is seen in a subset of dysplastic lesions of the cervix that fail to be detected by the Pap test, whereas normal patterns were observed in lesions that were Pap positive. These data have led to the hypothesis that there is a subset of lesions where dysplastic cells fail to exfoliate. In this model, the Pap test is extremely unlikely to detect such "nonshedding" lesions as the cells resist being harvested by the collection devices. However, despite their nonshedding behavior, these lesions remain readily identifiable by visual inspection of the cervix after application of acetic acid. PMID- 12634626 TI - Comparison of cell collection and direct visualization cervical cancer screening adjuncts. AB - Worldwide, invasive cancer of the cervix is the second most common malignancy in women. Each year, approximately 400,000 women are diagnosed with invasive cancer of the cervix, whereas 200,000 women die from the disease. In the United States, in 2002, there were 12,900 newly diagnosed cases of invasive cervical cancer and 4,400 cervical cancer-related deaths. Data from a recent meta-analysis found that the mean sensitivity of the conventional Papanicolaou (Pap) smear in screening populations was only 47%. These data, and others, underscore the need for more reliable and sensitive methods to screen for cervical cancer precursors. One example is the PapSure screening examination, at this point the only in-office direct visual cervical screening examination cleared by the US Food and Drug Administration. This cervical screening examination combines the traditional Pap smear with a direct visual screening test, speculoscopy (ie, visual inspection of the cervix with a chemiluminescent light, the Speculite). PMID- 12634628 TI - Coagulation or excision of ovarian endometriomas? AB - A systematic review was undertaken to determine whether coagulation or laser vaporization of endometriomas is associated with an increase in the risk of cyst recurrence compared with excision of the pseudocapsule. In the four comparative trials identified, endometrioma recurrence was reported in 39 of 212 (18.4%) women treated with coagulation or laser vaporization compared with 19 of 295 (6.4%) in those who underwent cystectomy. The odds ratios of endometrioma recurrence ranged from 1.41 to 9.38 with 95% CIs including unity in two studies. The common odds ratio was 3.09 (95% CI 1.78-5.36). Coagulation or laser vaporization of endometriomas without excision of the pseudocapsule seems to be associated with a significant increase in risk of cyst recurrence. PMID- 12634629 TI - Ovulation induction/intrauterine insemination in infertile couples is associated with low-birth-weight infants. AB - OBJECTIVE: The study was undertaken to determine the outcome of singleton pregnancies conceived through ovulation induction and intrauterine insemination using a partner's (OI/IUI) or donor sperm (OI/IUID) by comparison with naturally conceived singletons within a national cohort. STUDY DESIGN: Retrospective cohort study in primigravid women who were treated with OI/IUI (n = 97) or OI/IUID (n = 35) at Glasgow Royal Infirmary between March 1993 and March 1997 and identified within the Scottish national cohort (n = 109,443) delivering during the same period. Univariate and multivariate logistic regression models were used to examine outcomes and to determine the factors associated with infertility treatment and low birth weight. RESULTS: OI/IUI singletons were more likely to be lighter, of clinically low birth weight, and born more prematurely than natural conceptions. After adjustment for associated factors, these infants were 4.85 times (95% CI 2.25-10.48) more likely to be of low birth weight. We could not demonstrate any differences between OI/IUID and natural conceptions. CONCLUSION: The perinatal outcome of singletons born to subfertile mothers conceived through OI/IUI is poorer than that of matched natural conceptions. We suggest that intrinsic factors in subfertile couples predispose them to having smaller infants and that "infertility" should be added to the list of recognized factors associated with low birth weight. PMID- 12634630 TI - The benefit of preterm birth at tertiary care centers is related to gestational age. AB - OBJECTIVE: The purpose of this study was to examine the relationship between gestational age and outcomes of outborn versus inborn preterm infants. STUDY DESIGN: Multivariable logistic regression analysis was used to examine gestational age-specific, risk-adjusted outcomes of 2962 singleton infants who were born at <32 weeks of gestation who were admitted to 17 Canadian neonatal intensive care units from 1996 through 1997. RESULTS: The risk-adjusted incidence was significantly (P <.05) higher among outborn versus inborn infants for mortality rates (odds ratio, 2.2) and > or =grade 3 intraventricular hemorrhage (odds ratio, 2.1) at < or =26 weeks of gestation and for chronic lung disease (odds ratio, 1.7) at 27 to 29 weeks of gestation. Outcomes of outborn and inborn infants at 30 to 31 weeks of gestation were not significantly different. CONCLUSION: The short-term benefit of preterm birth at tertiary centers is related inversely to gestational age and may not extend beyond 29 weeks of gestation. PMID- 12634631 TI - Is Rh immune globulin needed in early first-trimester abortion? A review. AB - The prophylactic use of Rh immune globulin has been a medical success, protecting women who could be at risk from exposure to the Rh(D) antigen. Thus, it is not surprising that Rh(D) immunoprophylaxis has been extended from women with term pregnancies to all women with miscarriages, abortions, and ectopic pregnancies. In this article we review the existing medical literature to assess the risks of fetomaternal hemorrhage and Rh isoimmunization after complications of a first trimester pregnancy, induced abortion, or ectopic pregnancy. The evidence to support the use of Rh immune globulin in the first trimester is sparse, but there is theoretic evidence of its necessity. Despite weak evidence to support its use, there is little risk. PMID- 12634632 TI - Temporal and demographic trends in cerebral palsy--fact and fiction. AB - The rate of cerebral palsy has not decreased in developed countries over the past 30 years, despite the widespread use of electronic fetal heart rate monitoring and a 5-fold increase in the cesarean delivery rate over the same period of time. However, neonatal survival has improved during these decades. These observations have lead to the hypothesis that increased survival of premature, neurologically impaired infants may have masked an actual reduction in cerebral palsy among term infants as a result of the use of electronic monitoring and the avoidance of intrapartum asphyxia. A review of the medical literature, as well as a demographic analysis of term and preterm birth rates in the United States, refutes this hypothesis on four grounds. First, cerebral palsy prevalence has been separately analyzed in term infants and shows no change over 30 years. Second, the prevalence of cerebral palsy is the same or lower in underdeveloped countries than in developed nations; in the former, the availability of emergency cesarean delivery based on electronic monitor data is limited or absent. Third, the increase in prevalence of cerebral palsy among low-birth-weight infants and the increase in cesarean sections based on presumed fetal distress were not simultaneous events-the former preceded the latter by a decade. Improved neonatal survival since the 1980s has been associated with a stable or decreasing rate of neurologic impairment and thus could not have obscured improvement from reduced term asphyxia. Finally, compared with the number of infants born by cesarean section for fetal distress, there are simply not enough infants born in the most vulnerable weight groups to make any impact on even a minimal improvement of outcome in the group delivered by cesarean section for presumed fetal distress. Except in rare instances, cerebral palsy is a developmental event that is unpreventable given our current state of technology. PMID- 12634633 TI - Increased expression of cyclin G1 in leiomyoma compared with normal myometrium. AB - OBJECTIVE: The purpose of this study was to detect the expression of cyclin G1 in leiomyoma and to investigate the alteration of its expression compared with normal myometrial tissue that was obtained from the same patient. STUDY DESIGN: With the use of Northern blot analysis, Western blot analysis, and immunohistochemistry, we analyzed the expression of cyclin G1 in 24 patients who underwent hysterectomies. RESULTS: We found that messenger RNA levels of cyclin G1 were elevated in human leiomyomas compared with their adjacent normal myometrial tissues. Consistent with elevated messenger RNA levels, high levels of cyclin G1 protein expression were detected by immunoblot analysis in all leiomyoma samples. Immunohistochemistry revealed that cyclin G1 is located mainly in the nucleus in both normal myometrium and leiomyoma. However, higher levels of cyclin G1 were apparent in tumor regions compared with adjacent normal myometrial regions. In addition, we found the expression levels of other cyclins (A and E) and CDK2 were elevated in leiomyomas compared with normal myometrium. Because cyclin G1 is a transcriptional target of the p53 tumor suppressor, we examined the p53 status of all eight leiomyoma samples and found no p53 mutations. CONCLUSION: These results suggest that cyclin G1 is frequently overexpressed in uterine leiomyoma in a p53-independent manner and that this abnormality could be attributed to the severe proliferation of human uterine leiomyomas. PMID- 12634634 TI - Cervical adenocarcinoma survival among Hispanic and white women: a multicenter cohort study. AB - OBJECTIVE: We compared the clinical outcome of cervical adenocarcinoma in Hispanic and white women to determine whether race was an independent predictor of survival. STUDY DESIGN: All women who were diagnosed with cervical adenocarcinoma at three institutions between 1982 and 2000 were identified. Medical records were reviewed retrospectively. Hispanic and white cohorts were matched 1:2 for age, stage of disease, date of diagnosis, tumor size, histologic subtype, grade, and invasive depth. RESULTS: The 65 Hispanic patients were more likely to be treated at the public hospital (71% vs 14%; P <.001) than the 122 matched white patients. Most Hispanic patients (72%) and white patients (76%) presented with early (stage IA-IIA), not advanced (IIB-IVB), disease. Early (81% vs 81%, P =.65), advanced (37% vs 26%, P =.21), and overall 5-year survival rates (67% vs 68%, P =.57) were similar among Hispanic and white patients, respectively. The relative risk of race on recurrence was 1.22 (95% CI, 0.56 2.42) and on survival was 0.72 (95% CI, 0.36-1.44). CONCLUSION: Hispanic race is not an independent predictor of survival in cervical adenocarcinoma. PMID- 12634635 TI - Sacrospinous cervicocolpopexy with uterine conservation for uterovaginal prolapse in elderly women: an evolving concept. AB - OBJECTIVE: The purpose of this study was to evaluate the efficacy of the performance of sacrospinous cervicocolpopexy with uterine conservation in the treatment of uterovaginal prolapse in women over the age of 60 years. STUDY DESIGN: This was a nonrandomized prospective controlled study. Between February 1996 and January 2001, 109 women above the age of 60 years with a complaint of symptomatic uterovaginal prolapse were treated with either sacrospinous cervicocolpopexy with uterine conservation or vaginal hysterectomy concomitant with sacrospinous colpopexy. Patients were reviewed at 6 weeks and 6 and 12 months after operation and then yearly thereafter. Preoperative patient characteristics, operative, and postoperative events and follow-up results were recorded. RESULTS: During the study period, 61 patients (56%) underwent sacrospinous cervicocolpopexy with uterine conservation (group A), whereas 48 patients (44%) had vaginal hysterectomy performed concomitantly with sacrospinous colpopexy (group B). The mean age for the two groups was comparable (70.1 +/- 6 years vs 69.4 +/- 4.6 years, respectively; P =.8). Group A had significantly less blood loss (P <.01), shorter operating time (P <.01), and fewer complications after surgery (P =.01) compared with group B. After a mean follow-up period of 33 and 34 months, respectively, the two groups had comparable success rates with regard to uterine and upper vaginal support (93.5% and 95.9%, respectively; P =.6). Recurrent cystocoele developed in 11.4% and 10.4% of groups A and B, respectively (P =.9). Within the follow-up period, 3 patients (5%) in group A and 2 patients (4.2%) in group B underwent repeat operation for recurrent uterovaginal or vault prolapse. CONCLUSION: Sacrospinous cervicocolpopexy with uterine conservation is a safe and effective surgical option that could benefit elderly patients with uterovaginal prolapse. It avoids the potential morbidity of vaginal hysterectomy and is associated with a high success rate. PMID- 12634636 TI - Expression and regulation of interleukin-8 in human fallopian tubal cells. AB - OBJECTIVE: The human fallopian tube creates the microenvironment for fertilization and early embryogenesis. Salpingitis may result in infertility and ectopic pregnancy by causing tubal blockage and hydrosalpinx. To better understand the relationship between infectious inflammation and tubal damage, we investigated the expression and regulation of interleukin-8 in human tubal epithelial and stromal cells in culture. STUDY DESIGN: Human fallopian tube epithelial and stromal cell cultures were used to measure interleukin-8 messenger RNA and interleukin-8 protein levels at basal conditions and after stimulation with interleukin-1alpha and tumor necrosis factor-alpha. Northern blot analysis and enzyme-linked immunosorbent assay were used to evaluate messenger RNA and protein levels, respectively. RESULTS: Tubal epithelial cells expressed high levels of interleukin-8 messenger RNA and secreted significantly more immunoreactive interleukin-8 into culture medium than did tubal stromal cells (2065 +/- 153 pg/mg vs 530 +/- 56 pg/mg of total protein, P <.01). Interleukin 1alpha and TNF-alpha treatments induced a concentration-dependent increase in interleukin-8 messenger RNA expression in both epithelial and stromal cells. However, at the protein level, although interleukin-1alpha and tumor necrosis factor-alpha treatments increased the secretion of interleukin-8 from stromal cells significantly, similar treatments had no effect on interleukin-8 secretion from epithelial cells. CONCLUSION: The expression of interleukin-8 in human tubal epithelial and stromal cells is different. Interleukin-8 expression of tubal epithelial and stromal cells in response to inflammatory cytokines such as interleukin-1alpha and tumor necrosis factor-alpha also varies. This may be important in the pathogenesis of salpingitis. PMID- 12634637 TI - Comparison of human papillomavirus genotypes, sexual, and reproductive risk factors of cervical adenocarcinoma and squamous cell carcinoma: Northeastern United States. AB - OBJECTIVE: Although human papillomavirus causes essentially all cervical carcinoma, cofactors may differ by cancer histologic type. We examined human papillomavirus genotypes and sexual and reproductive risk factors for cervical adenocarcinoma and squamous cell carcinoma. STUDY DESIGN: One hundred twenty-four women with adenocarcinoma, 139 women with squamous cell carcinoma, and 307 control subjects participated in this case-control study. Logistic regression analyses were performed to calculate odds ratios and CIs. RESULTS: Human papillomavirus 18 was associated most strongly with adenocarcinoma (odds ratio, 105; 95% CI, 23-487). Human papillomavirus 16 was associated most strongly with squamous cell carcinoma (odds ratio, 30; 95% CI, 12-77). More than three lifetime sexual partners was a risk factor for adenocarcinoma (odds ratio, 2.1; 95% CI, 1.1-4.0) and squamous cell carcinoma (odds ratio, 3.0; 95% CI, 1.6-5.9). Even being pregnant was associated inversely with adenocarcinoma (odds ratio, 0.4; 95% CI, 0.2-0.8). Five or more pregnancies was associated with squamous cell carcinoma (odds ratio, 2.2; 95% CI, 0.9-5.4). CONCLUSION: The relative importance of human papillomavirus genotypes 16 and 18 and the reproductive co-factor differences suggest distinct causes for cervical adenocarcinoma and squamous cell carcinoma. PMID- 12634638 TI - Mifepristone and misoprostol and methotrexate/misoprostol in clinical practice for abortion. AB - OBJECTIVE: The purpose of this study was to evaluate the efficacy, side-effect profile, and follow-up rates in women who obtain a medical abortion in a nonresearch setting. STUDY DESIGN: From December 1, 2000, to June 30, 2001, we prospectively followed 218 women who had been evaluated in our private office for medical abortion. Women received either mifepristone 200 mg orally followed 1 to 2 days later by self-administered misoprostol 800 microg vaginally or methotrexate 50 mg/m(2) intramuscularly followed 3 to 7 days later by self administered misoprostol 800 microg vaginally. RESULTS: Of the 174 women who had a medical abortion, 148 women (85%) chose mifepristone/misoprostol, and 26 women (15%) chose methotrexate/misoprostol. In women up to 49 days of gestation, complete abortion occurred by the first follow-up visit in 82 of 86 women (95%; 95% CI, 89-99) and in 21 of 25 women (84%; 95% CI, 64-95) women, respectively. In women who used mifepristone/misoprostol from 50 to 63 days of gestation, complete abortions occurred in 56 of 59 women (95%; 95% CI, 86-99) women. Four women (2%; 95% CI, 1-6) were lost to follow-up. CONCLUSION: Medical abortion with mifepristone/misoprostol and with methotrexate/misoprostol can be provided in a nonresearch setting with efficacy similar to that reported in the medical literature for research protocols. PMID- 12634639 TI - Dipeptidyl peptidase IV expression in endometrial endometrioid adenocarcinoma and its inverse correlation with tumor grade. AB - OBJECTIVES: Dipeptidyl peptidase IV (DPPIV)/CD26 is a cell surface aminopeptidase. This study investigated the expression and localization of DPPIV in endometrial endometrioid adenocarcinomas of different grades. STUDY DESIGN: Immunohistochemical analysis was performed by using DPPIV and regulated on activation, normal T-cell expressed and secreted (RANTES) specific monoclonal antibodies. Cell proliferation was evaluated by bromodeoxyuridine (BrdU) uptake assay. RESULTS: Immunohistochemical analyses showed that DPPIV was strongly or moderately stained in glandular cells of the normal secretory phase. In endometrial adenocarcinoma, the DPPIV expression decreased with advancing grade (P <.01). Furthermore, RANTES, one of the possible DPPIV substrates, was highly expressed in all grades of endometrial adenocarcinoma cells. The addition of RANTES to endometrial adenocarcinoma cells increased proliferation in a concentration-dependent manner. CONCLUSION: DPPIV is expressed in normal endometrial glandular cells, but its expression in endometrial adenocarcinoma is down-regulated with increasing grade. Our data also suggest a regulatory role of this ectoenzyme in neoplastic transformation and progression of endometrial adenocarcinomas possibly by degrading certain bioactive peptides such as RANTES. PMID- 12634640 TI - Clinical findings among young women with genital human papillomavirus infection. AB - OBJECTIVE: The purpose of this study was to identify clinical signs and symptoms associated with detection of human papillomavirus (HPV) DNA in the female genital tract. STUDY DESIGN: A total of 516 university students (18 to 24 years old) enrolled in a cohort study that included the collection of genital specimens for HPV DNA testing every 4 months for up to 4 years. Reported symptoms and objective clinical findings of women with and without HPV DNA were compared by multivariate analysis. RESULTS: Acute and persisting HPV infections were not associated with discharge, itching, burning, soreness, or fissures. Clinical evidence of genital warts was statistically associated only with HPV types 6 and 11. Detection of any HPV DNA was associated with bacterial vaginosis (BV). Furthermore, a time lag analysis suggests that HPV infection usually precedes detection of BV. CONCLUSION: Most women who acquire genital HPV infection are asymptomatic; some, however, are at increased risk for BV. PMID- 12634641 TI - A new scoring system to differentiate benign from malignant adnexal masses. AB - OBJECTIVE: The purpose of this study was to develop and cross-validate a new sonographic scoring system for differentiation between benign and malignant adnexal masses. STUDY DESIGN: This study was conducted in a tertiary care university hospital. In the first part of the study, we used a multivariate logistic regression analysis to develop a scoring system that was based on morphologic and Doppler sonographic data for 705 adnexal masses in 665 patients who were diagnosed and treated at our institution from January 1995 to June 2001. The scoring system was designed to use only those parameters that are found to be independent predictors of malignancy. In the second part of the study, we prospectively cross-validated this scoring system in a series of 90 adnexal masses in 86 patients between July 2001 and March 2002. With the use of the area under the curve of the respective ROC curves, we compared the new scoring system with other scoring systems. RESULTS: Multivariate logistic regression analysis revealed that the only independent predictor parameters were thick papillary projections, solid areas, central flow, and velocimetric features of high velocity and low resistance. In the prospective cross-validation study, our scoring system had the best diagnostic performance (area under the curve, 0.98) compared with Sassone (area under the curve, 0.89; P =.017), De Priest (area under the curve, 0.92; P =.048), and Ferrazzi (area under the curve, 0.90; P =.013) scoring systems. CONCLUSION: Our new sonographic scoring system had a better diagnostic performance than three previously published scoring systems. PMID- 12634642 TI - Treatment of women with low-grade squamous intraepithelial lesions on cytologic evidence or biopsy results by board-certified gynecologists. AB - OBJECTIVE: The Bethesda system combined human papillomavirus effects and mild dysplasia/cervical intraepithelial neoplasia grade 1 into a single category called low-grade squamous intraepithelial lesion. We sought to determine whether the treatment of women with low-grade squamous intraepithelial lesions by board certified gynecologists was different for human papillomavirus effect versus mild dysplasia/cervical intraepithelial neoplasia grade 1. In addition, we sought to document the prevalence of human papillomavirus testing and counseling regarding sexual partners by this group of providers. STUDY DESIGN: A 2-page survey was mailed in April 1998 to a random sample of 491 fellows of the American College of Obstetrics and Gynecology. The response rate was 50.6%. RESULTS: For a Papanicolaou test with human papillomavirus effects, 52% of the respondents would perform colposcopy. For a Papanicolaou test with mild dysplasia/cervical intraepithelial neoplasia 1, 86% of the respondents would perform colposcopy (P <.0001). For biopsy-proved condyloma, 33% of the respondents would treat the patient. For biopsy-proved mild dysplasia/cervical intraepithelial neoplasia grade 1, significantly more respondents would treat the patient (45%, P =.02). Respondents rarely offered routine human papillomavirus testing and frequently made recommendations regarding partner examination and condom use. CONCLUSION: Board-certified gynecologists commonly treat women with a newly diagnosed low grade squamous intraepithelial lesion Papanicolaou test with colposcopy and frequently differentiate between human papillomavirus effects and mild dysplasia/cervical intraepithelial neoplasia grade 1 in the treatment. PMID- 12634643 TI - Durability of treatment effects after endometrial cryoablation versus rollerball electroablation for abnormal uterine bleeding: two-year results of a multicenter randomized trial. AB - In a randomized trial, 93% of cryoablation patients who were evaluated at both 12 and 24 months and not retreated were free of abnormal uterine bleeding at 12 months and 94% at 24 months versus 92% of electroablation patients at both times. The retreatment rate was similar after cryoablation (12.9%) and electroablation (14.0%). PMID- 12634644 TI - Immunohistochemical detection of insulin-like growth factor type I receptor and uterine volume changes in gonadotropin-releasing hormone analog-treated uterine leiomyomas. AB - OBJECTIVE: This study was undertaken to assess the relationship between insulin like growth factor (IGF) type I receptor (IGF-I-R) expression in uterine leiomyomas after gonadotropin-releasing hormone (GnRH) analog administration and modifications in uterine size. STUDY DESIGN: Forty-six women with menorrhagia for uterine leiomyomatosis were treated monthly with leuprolide acetate depot 3.75 mg before undergoing surgery. The uterine volume before and after therapy was assessed by transabdominal ultrasonography. Immunohistochemical detection of IGF I-R was performed on leiomyoma tissue samples. The relationship between IGF-I-R levels and uterine volume changes was analyzed. RESULTS: Uterine volume decreased after therapy. Patients with a lower immunohistochemical expression of IGF-I-R showed a larger decrease in uterine size. CONCLUSION: The shrinkage in uterine volume induced by GnRH analogs seems to be related to the observed reduction in IGF-I-R levels. So, the IGF-I/IGF-I-R system might be involved in leiomyoma growth, and the pharmacologic action of GnRH analogs on uterine leiomyomas might also be related to the effects on IGF-I-R expression. PMID- 12634645 TI - Differing symptoms of abused versus nonabused women in obstetric-gynecology settings. AB - OBJECTIVE: The purpose of this study was to document the prevalence rates of reported abuse in obstetric/gynecology settings and to characterize physical symptoms, function, and psychologic distress of women who report abuse. STUDY DESIGN: This was a descriptive study of 118 patients who reported physical or sexual abuse in the past year and 2800 patients who reported no abuse at seven obstetric/gynecology practices. The Patient Health Questionnaire measured psychiatric diagnoses, somatic complaints, and psychosocial stressors. The Medical Outcomes Study Short Form 20 assessed health status, and self-reported health care use was measured. RESULTS: Four percent of women reported recent abuse. Patients who reported abuse had more psychiatric diagnoses, more severe psychologic distress, more somatic symptoms, more psychosocial stressors, more physician visits, and poorer reported health status compared with nonreporters of abuse. CONCLUSION: Enhanced recognition of possible abuse through screening may identify women in need of intervention and help to reduce the need for continued medical intervention. PMID- 12634646 TI - Maternal and transplacental pharmacokinetics of azithromycin. AB - OBJECTIVE: The purpose of this study was to characterize the pharmacokinetics of orally administered azithromycin in the term gravid woman. STUDY DESIGN: Twenty women who were scheduled for elective cesarean delivery were enrolled prospectively and received 1 g of oral azithromycin at either 6, 12, 24, 72, or 168 hours before the operation. All women received spinal anesthesia, at which time a sample of cerebrospinal fluid was obtained for analysis. Maternal serum and urine were obtained immediately before the operation. Intraoperatively, samples of myometrium, maternal adipose tissue, placenta, amniotic fluid, and umbilical arterial and venous cord blood were obtained. Azithromycin levels were determined quantitatively with high-pressure liquid chromatography with electrochemical detection. RESULTS: All participants tolerated the preoperative azithromycin without significant adverse reactions. Peak maternal serum azithromycin levels occurred within 6 hours of drug administration. Although high serum levels of azithromycin were reached early, a rapid decline in drug concentration was noted over the initial 24 hours after the drug administration (6-hour: 311 ng/mL; 24-hour: 63 ng/mL). In contrast, azithromycin levels in myometrial, adipose, and placental tissue were higher (>500 ng/mL) and sustained for up to 72 hours after administration. High urine levels of azithromycin (>5000 ng/mL) were noted similarly during the initial 72 hours after drug administration. Umbilical arterial and venous serum azithromycin levels were low (19-38 ng/mL) during the first 72 hours. Amniotic fluid levels were highest at 6 hours (151 ng/mL) and declined rapidly. Maternal cerebrospinal azithromycin concentrations were undetectable for all time points. CONCLUSION: Azithromycin has a rapid serum half-life in the term gravid woman with a prolonged half-life and high-sustained antibiotic levels noted within myometrium, adipose, and placental tissue. Given the broad antimicrobial spectrum and placental penetration, azithromycin may have potential use for the treatment of perinatal infections. PMID- 12634647 TI - Decreased tryptophan catabolism by placental indoleamine 2,3-dioxygenase in preeclampsia. AB - OBJECTIVE: Tryptophan degradation and depletion resulting from activation of indoleamine 2,3-dioxygenase is characteristic of inflammatory reactions and may control their intensity. Normal third-trimester pregnancy is characterized by a maternal systemic inflammatory response, which is more intense in preeclampsia. Therefore, we studied tryptophan metabolism in pregnant women, with or without preeclampsia, as well as expression and function of placental indoleamine 2,3 dioxygenase. STUDY DESIGN: Plasma concentrations of tryptophan and kynurenine in women with preeclampsia, appropriately matched women with normal pregnancy, and healthy nonpregnant women were measured. Placental enzymatic activity and messenger RNA (mRNA) expression level of indoleamine 2,3-dioxygenase were determined from the same placental material. Peripheral blood mononuclear cell proliferation was determined in medium conditioned by prior culture with villous tissue. RESULTS: The plasma ratio of kynurenine to tryptophan, an in vivo index of enzyme activity, was significantly increased compared with nonpregnant controls in normal pregnancy but not in preeclampsia. The activity and mRNA expression level of indoleamine 2,3-dioxygenase in term placentas were significantly lower in preeclampsia. Medium conditioned by culture of villous tissue explants of preeclampsia was less effective in inhibiting peripheral blood mononuclear cell proliferation compared with that of normal pregnancy. CONCLUSION: These observations suggest that in preeclampsia, reduced placental indoleamine 2,3-dioxygenase activity (and relatively elevated plasma tryptophan) could cause dysregulation of the inflammatory response that is intrinsic to normal pregnancy. This may contribute to the pathogenesis of the maternal syndrome of preeclampsia. PMID- 12634648 TI - Ductus venosus flow velocities in relation to the cardiac defects in first trimester fetuses with enlarged nuchal translucency. AB - OBJECTIVE: Our aim is to study first-trimester ductus venosus (DV) velocities in relation to cardiac abnormalities. STUDY DESIGN: Ultrasound examination was performed in 85 normal fetuses and 45 fetuses with a nuchal translucency (NT) >95th percentile. Pulsatility index for veins (PIV) and velocity during late diastole (a-V) of the DV were measured and compared with the use of multilevel analysis, between fetuses with a heart defect and those without. RESULTS: Compared with 85 normal fetuses, the a-V and PIV of the fetuses with NT>95th percentile and normal hearts were decreased and increased, respectively. The a-V and PIV of 11 fetuses with NT>95th percentile and cardiac defects were decreased and increased compared with the 25 fetuses with normal hearts, irrespective of the karyotype. The most common cardiac malformations were septal defects. CONCLUSION: Fetuses with NT >95th percentile and cardiac defects show markedly changed DV velocities. Because the type of cardiac defects cannot always explain the hemodynamic changes found in these fetuses, some other mechanisms seem to be involved. PMID- 12634649 TI - Uterine contractility in women whose fetus is delivered in the occipitoposterior position. AB - OBJECTIVE: Most fetuses in the occipitoposterior position rotate spontaneously after striking the pelvic floor. The increased prevalence of prolonged labor, operative delivery, and oxytocin augmentation in women with an occipitoposterior fetal position seems consistent with decreased uterine contractility. We sought to test the hypothesis that women with a persistent occipitoposterior fetal position have inadequate intrauterine pressure. STUDY DESIGN: Intrauterine pressure was measured prospectively electronically in 94 women whose labor pain was controlled by patient-requested epidural analgesia. Eleven women (12%) were delivered as a persistent occipitoposterior fetal position. In a nested case control study, these women were compared with 22 women who were delivered as an occipitoanterior fetal position who were matched for age, parity, gestational age, cervical examination at study enrollment, and body mass index. The intrauterine pressure measurements were initiated during the first stage of labor and continued throughout the entire labor process. Women were encouraged in the second stage of labor, after a period of recording baseline contractility, to push using a standardized Valsalva maneuver once the vertex reached the +2 station. The area under the intrauterine pressure curve (integral) was used to estimate uterine contractility and expulsive performances. RESULTS: Five women (45%) in the occipitoposterior group required operative delivery. The average duration of the second stage of labor in the occipitoposterior group was 91.4 +/- 23.2 minutes compared with 51.7 +/- 6.6 minutes in the occipitoanterior fetal position (P =.04). Ninety percent of women in the occipitoposterior group required oxytocin, compared with 59% of the women in the occipitoanterior group (P =.11). There were no differences in uterine contractility between occipitoposterior and occipitoanterior groups during either the first stage of labor (integral mean +/- SEM: occipitoposterior [1685.3 +/- 194.6 mm Hg. s] vs occipitoanterior fetal position [1700.8 +/- 128.9 mm Hg. s, P =.98]) or second stages of labor (occipitoposterior [1952.6 +/- 186.5 mm Hg. s] vs occipitoanterior fetal position [1740.8 +/- 104.3 mm Hg. s, P =.46]). Further, there were no significant differences in pushing performances between the occipitoposterior and occipitoanterior groups (Valsalva maneuver: occipitoposterior 2864.9 +/- 328.8 mm Hg. s] vs occipitoanterior [2898.6 +/- 222.2 mm Hg. s, P =.90]). CONCLUSION: Women who were delivered as a persistent occipitoposterior fetal position do not have lower intrauterine pressure levels immediately before or during the second stage of labor. PMID- 12634650 TI - Increased vascular endothelial cell production of interleukin-6 in severe preeclampsia. AB - OBJECTIVE: The purpose of this study was to determine whether plasma from women with severe preeclampsia stimulates the production of endothelial cell interleukin-6 production and whether vitamin E could inhibit this process. STUDY DESIGN: Human umbilical vein endothelial cells were cultured in the presence of 5% plasma from women with severe preeclampsia (n = 12) or healthy pregnant women at term (n = 12), with or without 50 micromol/L vitamin E. Levels of interleukin 6 in plasma and human umbilical vein endothelial cell-conditioned media were measured by enzyme-linked immunosorbent assay. RESULTS: Interleukin-6 levels were elevated 5-fold in preeclamptic plasma compared with normal pregnant plasma (P <.05). Human umbilical vein endothelial cell interleukin-6 production was increased 25% by preeclamptic plasma compared with normal pregnant plasma (P <.005), and increased interleukin-6 production by preeclamptic plasma was inhibited by vitamin E. CONCLUSION: Endothelial cell activation by preeclamptic plasma stimulates interleukin-6 production, which is inhibited by vitamin E. These findings provide a potential cellular mechanism for the beneficial effects of antioxidant therapy in preeclampsia. PMID- 12634651 TI - Combined first-trimester versus second-trimester serum screening for Down syndrome: a cost analysis. AB - OBJECTIVE: The purpose of this study was to compare the cost-effectiveness of combined first-trimester screening for fetal Down syndrome with second-trimester maternal serum triple screening. STUDY DESIGN: A first-trimester screening approach that used nuchal translucency measurement and maternal serum screening was evaluated against second-trimester maternal serum triple screening in a hypothetic population. Screening sensitivities and screen-positive rates were 91% and 5% for the first-trimester approach and 70% and 7.5% for the second-trimester approach, respectively. The costs of fetal Down syndrome, live-born Down syndrome cost, and total costs (screening plus live-born costs) were calculated for each screening program. RESULTS: First-trimester screening was associated with lower screening and live-born Down syndrome costs versus second-trimester serum screening. Total Down syndrome screening costs were 29.1% lower with first trimester screening. CONCLUSION: In this hypothetic model, combined first trimester screening for fetal Down syndrome was more cost-effective than universal second-trimester triple serum screening. PMID- 12634652 TI - Antibiotic treatment of bacterial vaginosis in pregnancy: a meta-analysis. AB - OBJECTIVE: The purpose of this study was to evaluate the effectiveness of antibiotic treatment of bacterial vaginosis in pregnancy to reduce preterm delivery. STUDY DESIGN: We performed a meta-analysis of published, English language, randomized, placebo-controlled clinical trials of antibiotic treatment of bacterial vaginosis in pregnant women with intact amniotic membranes at <37 weeks of gestation. Primary outcomes included preterm delivery, perinatal or neonatal death, and neonatal morbidity. RESULTS: Ten studies with results for 3969 patients were included. In patients without preterm labor, antibiotic treatment did not significantly decrease preterm delivery at <37 weeks of gestation, in all patients combined (odds ratio, 0.83; 95% CI, 0.57-1.21) nor in high-risk patients with a previous preterm delivery (odds ratio, 0.50; 95% CI, 0.22-1.12). In both groups, significant statistical heterogeneity was observed. A significant reduction in preterm delivery and no statistical heterogeneity were observed in 338 high-risk patients who received oral regimens with treatment durations of > or =7 days (odds ratio, 0.42; 95% CI, 0.27-0.67). Nonsignificant effects and no statistical heterogeneity were observed in low-risk patients (odds ratio, 0.94; 95% CI, 0.71-1.25) and with vaginal regimens (odds ratio, 1.25; 95% CI: 0.86-1.81). In one study antibiotic treatment in patients with preterm labor led to a nonsignificant decrease in the rate of preterm deliveries (odds ratio, 0.31; 95% CI, 0.03-3.24). CONCLUSION: The screening of pregnant women who have bacterial vaginosis and who have had a previous preterm delivery and treatment with an oral regimen of longer duration can be justified on the basis of current evidence. More studies are needed to confirm the effectiveness of this strategy, both in high-risk patients without preterm labor and in patients with preterm labor. PMID- 12634654 TI - Fused umbilical arteries near placental cord insertion. AB - OBJECTIVE: This study was undertaken to study the frequency and cause of fused umbilical arteries (FUAs). STUDY DESIGN: Umbilical cord on the placental side was carefully examined for FUAs by continuous interval sections. RESULTS: The frequency was 3.1% or 22 in 702 infants with the frequency in female infants of 4.1% being significantly higher than that in male infants of 2.0%. The mean distance of fused points was 2.3 +/- 1.6 cm from placental cord insertion. Five of 22 cases showed both arterial fusion and division in the same umbilical cord. Marginal and velamentous insertions were significantly associated with FUAs. Gestational age, birth weight, and placental weight of 22 cases were within normal range. CONCLUSION: Single umbilical artery should be confirmed by 2 or 3 sections of umbilical cord because of the high frequency of FUA on the placental side. PMID- 12634653 TI - Changes in the isoforms of the sodium pump in the placenta and myometrium of women in labor. AB - OBJECTIVE: We determined whether changes in sodium pump isoform abundance accompanied active human labor. STUDY DESIGN: Specimens of placenta, amniochorion, and myometrium were collected from women in active spontaneous labor and from those not in labor. The abundance of the three sodium pump alpha isoforms was determined by Western blot analysis. RESULTS: Levels of the alpha1 and alpha2 isoforms were comparable in the three tissues for women in labor and not in labor. However, alpha3 isoform abundance in placenta and myometrium (but not amniochorion) was significantly decreased in women in active labor compared with women not in labor (sodium pump alpha3 in placenta: no labor 91.2 +/- 27.6 vs labor 46.9 +/- 3.6 density units, P =.002. Sodium pump alpha3 in myometrium: no labor 52.3 +/- 7.7 vs labor 19.8 +/- 1.6 density units, P =.0002). CONCLUSION: Because reductions in sodium pump number can result in hormone release from secretory tissues and in contraction of muscle, this suggests that the sodium pump may play a significant role in the initiation or maintenance of human labor. PMID- 12634655 TI - Effects of hyperthermia on hypoxic-ischemic brain damage in the immature rat: its influence on caspase-3-like protease. AB - OBJECTIVE: Recent clinical studies suggested that intrapartum maternal fever is a strong independent risk factor for neonatal encephalopathy. With use of a well studied rat model of neonatal hypoxic-ischemic encepalopathy, this study investigated the hypothesis that intraischemic hyperthermia accelerates and worsens brain injury in immature animals and examined whether apoptotic cell death machinery is involved in the underlying mechanisms. STUDY DESIGN: Seven-day old rats underwent a combination of left common carotid artery ligation and exposure to 8% oxygen for 15 minutes (n = 32 rats). During the 15-minute hypoxic insult, body temperature was elevated to 40 degrees C in 16 animals (hyperthermic hypoxic insult group), and was maintained at 37 degrees C in 16 animals (normothermic hypoxic insult group). Then both groups were placed in the same chamber in a water bath at 37 degrees C for 24 hours and finally returned to the mothers. Caspase-3-like activity was assessed 36 hours after the hypoxic-ischemic insult. One week later, microtubule-associated protein-2 immunostaining was used to examine neuronal damage. RESULTS: Intraischemic hyperthermia was shown to activate the caspase-3 activity 36 hours after hypoxia-ischemia while caspase-3 was activated insignificantly in the normothermic hypoxic insult group at that time. The hyperthermic hypoxic insult group also showed a reduced microtubule associated protein-2-positive area 7 days after hypoxia-ischemia compared with that in the normothermia group. CONCLUSION: Hyperthermia during hypoxia-ischemia makes the immature brain inordinately susceptible to hypoxic-ischemic insult and causes brain injury, even if hypoxic-ischemic insult is so mild that it causes no or little injury by itself. This effect may be mediated by the escalation of the apoptotic cell death pathway in the immature animal. PMID- 12634657 TI - Nitric oxide metabolites in cervical fluid during pregnancy: further evidence for the role of cervical nitric oxide in cervical ripening. AB - OBJECTIVE: Cervical tissue expresses all the isoenzymes of nitric oxide synthase. We studied the concentrations of nitric oxide metabolites in the cervical fluid in nonpregnant (n = 11) and pregnant women (n = 106). STUDY DESIGN: Cervical fluid was collected into a Dacron polyester swab, and nitric oxide metabolites were eluted into physiologic saline solution, which was assayed for nitric oxide metabolites with the Griess reaction. The detection limit of the method is 0.2 micromol/L. RESULTS: Cervical fluid nitric oxide metabolite was detectable in 46% of nonpregnant women (median, <0.2 micromol/L; 95% CI, 0-49), in 63% of women in early pregnancy (median, 11 micromol/L; 95% CI, 0-23) and in 82% of women in late pregnancy (median, 128 micromol/L; 95% CI, 21-276). In late pregnancy, the cervical fluid nitric oxide metabolite level was higher in women with Bishop score of > or =6 (median, 163 micromol/L; 95% CI, 105-276) than in women with Bishop score of <6 (median, 86 micromol/L; 95% CI, 21-99). Cervical fluid nitric oxide metabolite concentration before the onset of labor in parous women (median, 97 micromol/L; 95% CI, 78-283) was higher (P =.008) than that in nulliparous women (median, 28 micromol/L; 95% CI, 0-95). Cervical fluid nitric oxide metabolites before the initiation of labor (median, 33 micromol/L; 95% CI, 0-95) rose to 3.5-fold (median, 115 micromol/L; 95% CI, 78-284) after the commencement of uterine contractions and showed a significant relationship to Bishop score (r = 0.39, P =.01). Cervical fluid nitric oxide metabolite concentrations were not relative to simultaneous plasma nitric oxide metabolite levels (n = 41 women, r = 0.14, P =.41). Rupture of fetal membranes tended to decrease cervical fluid nitric oxide metabolite levels, whereas gentle cervical manipulation elevated it 6.6-fold in 1 minute. The administration of glyceryl trinitrate (0.5 mg, nitric oxide donor) intracervically resulted in a significant rise in the cervical fluid nitric oxide metabolite level in 2 minutes. CONCLUSION: Cervical fluid nitric oxide metabolite level rises after cervical ripening, nitric oxide donor administration, or cervical manipulation, which supports a role for cervical nitric oxide in cervical ripening. PMID- 12634656 TI - Effect of histamine on phasic and tonic contractions of isolated uterine tissue from pregnant women. AB - OBJECTIVE: To enhance our understanding of the uterotonic effect of histamine, we compared the effects of histamine on spontaneous phasic and tonic contractile activity of uterine strips from term pregnant nonlaboring women. STUDY DESIGN: Longitudinal uterine strips were used from the lower uterine segment of term pregnant nonlaboring women undergoing elective cesarean section. The concentration-response relationship to histamine (10(-8) to 10(-4) mol/L) was determined in spontaneously contracting strips or in strips contracted tonically with a protein kinase C activator (-)-indolactam V in the presence of H(1) receptor antagonist (S[+]-chlorpheniramine maleate), H(2) receptor antagonist (cimetidine), or solvent. RESULTS: Histamine increased spontaneous phasic myometrial contractions in a concentration-dependent manner. H(1), but not H(2), receptor antagonist significantly attenuated the response to histamine. Histamine significantly reduced tonic contractions of uterine strips induced by indolactam V. H(1) histamine receptor antagonist abolished relaxation, whereas H(2) histamine receptor antagonist had no effect. CONCLUSION: Histamine increases spontaneous, but inhibits tonic, contractions of uterine strips from term pregnant nonlaboring women. Both effects are mediated through activation of H(1) receptors. PMID- 12634659 TI - Doppler assessment of the fetal cerebral hemodynamic response to moderate or severe maternal anemia. AB - OBJECTIVE: The purpose of this study was to evaluate the fetal vascular adaptation to moderate and severe maternal anemia. STUDY DESIGN: Biometry; amniotic fluid index; uterine, cerebral, and umbilical Doppler; and maternal hemoglobin level were measured at admission and 8 days after treatment. RESULTS: Group 1 consisted of 16 pregnancies (maternal hemoglobin level, 6.9 +/- 0.6 g/100 mL); group 2 consisted of 23 pregnancies (maternal hemoglobin level, 5 +/- 0.6 g/100 mL). At admission the cerebral and cerebral/umbilical Doppler indexes, amniotic index, and biometry were lower in group 2. The uterine index was normal in both groups. An abnormal fetal heart rate was found in group 2 only (48%). At day 8, maternal hemoglobin level and amniotic index increased more in group 2 than in group 1. The cerebral index and the cerebral-to-umbilical resistance ratio increased only in group 2. The abnormal fetal heart rate disappeared in group 2. CONCLUSION: Only severe maternal anemia (maternal hemoglobin level, <6 mg/L) triggered fetal cerebral vasodilation and reduced amniotic volume. PMID- 12634658 TI - Regulation of amniotic fluid volume by intramembranous absorption in sheep: role of passive permeability and vascular endothelial growth factor. AB - OBJECTIVE: During long-term intravascular fluid infusion in the ovine fetus, a large increase in fetal urinary flow rate occurs while amniotic fluid volume increases only slightly because of increased intramembranous absorption. The current study tested the hypotheses that passive intramembranous permeability increases in response to fetal intravascular saline solution infusion and that the increased intramembranous absorption occurs in parallel with an increase in vascular endothelial growth factor gene expression in the amnion, chorion, and placenta. STUDY DESIGN: Chronically catheterized fetal sheep that average 126 +/- 1 (SE) days of gestation either were infused intravascularly with 7 L of normal saline solution over 3 days (n = 8 sheep) or served as time controls (n = 6 sheep). Amniotic fluid volume and fetal urinary flow rate were measured daily. Intramembranous diffusional permeability was estimated daily as being equal to the clearance of intra-amniotically injected technetium 99m. Vascular endothelial growth factor messenger RNA abundance in the amnion, chorion, and placenta was determined by Northern blot analysis. Statistical analyses included analysis of variance. RESULTS: In the infused fetuses, amniotic fluid volume and urinary flow increased (P <.01) by 891 +/- 144 mL and 3488 +/- 487 mL per day, respectively, on infusion day 3 compared with no changes over time in the control fetuses. In the infused fetuses, estimated intramembranous absorption increased by 4276 +/- 499 mL during the 3-day infusion. Intramembranous technetium 99m permeability was similar over time in the two groups. In the infused group, vascular endothelial growth factor messenger RNA levels in the amnion, chorion, and placenta increased 2- to 4-fold compared with the control group (P <.001). CONCLUSION: The up regulation of vascular endothelial growth gene expression may mediate the increase in the intramembranous absorption that is induced by volume-loading diuresis; however, this does not occur by passive mechanisms. We speculate that vascular endothelial growth mediates the increased intramembranous absorption by increasing vesicular transport. PMID- 12634660 TI - The effects of community-acquired pneumonia during pregnancy ending with a live birth. AB - OBJECTIVE: The incidence and outcomes for community-acquired pneumonia during a pregnancy ending with a live birth in the province of Alberta, Canada, were analyzed. STUDY DESIGN: Hospital abstracts and vital statistics data between April 1, 1994, to March 31, 1999, were used. RESULTS: The incidence of hospitalization for community-acquired pneumonia was 1.51 (95% CI 1.35-1.67) per 1000 pregnancies in mothers 18 years and older compared with 1.47 (95% CI 1.42 1.51) per 1000 women (18 to 49 years) in the general population. The relative risk of staying longer at hospital for pneumonia was 0.44 during pregnancy compared with nonpregnant mothers with pneumonia. The relative risk of staying longer at hospital during delivery was 1.88, higher in those with a past hospitalization for community-acquired pneumonia. The relative risk of a small for-gestational-age newborn infant was 1.86 for those with a past hospitalization for community-acquired pneumonia compared with those without a past hospitalization. CONCLUSION: Community-acquired pneumonia during most pregnancies ending with a live birth appears to be well tolerated by mothers and newborn infants. PMID- 12634661 TI - Maternal serum activin A is not elevated before preeclampsia in women who are at high risk. AB - OBJECTIVE: The purpose of this study was to determine whether, in women who are at high risk of the development of preeclampsia, serum activin A concentrations are elevated before the disease and whether activin A is a useful predictor of preeclampsia. STUDY DESIGN: Sera were collected on five occasions throughout pregnancy from women with chronic hypertension, renal disease, or previous early onset preeclampsia (n = 80 women). Women were classified as control subjects (normotensive or stable chronic hypertension), gestational hypertensive, or preeclamptic (de novo or superimposed). Serum activin A concentrations were measured by immunoassay. Differences in activin A concentrations between groups were analyzed with the use of a mixed-models procedure; screening test characteristics were calculated. RESULTS: Twenty-six women (33%) had gestational hypertension, and 17 women (21%) had preeclampsia or superimposed preeclampsia. Serum activin A levels increased with gestation in all groups (P =.0001), but there were no significant difference in activin A levels between groups (P =.75). CONCLUSION: In women who were at high risk of the development of preeclampsia, serum activin A levels are not elevated with preeclampsia. Activin A is not a useful predictor of preeclampsia in this setting. PMID- 12634662 TI - Birth outcomes after prenatal exposure to antidepressant medication. AB - OBJECTIVE: The purpose of this study was to examine prospectively the incidence of congenital anomalies and neonatal complications after prenatal exposure to antidepressant medication. STUDY DESIGN: Birth outcomes were obtained from a review of obstetric and neonatal records of 138 women who were treated with selective serotonin reuptake inhibitor antidepressant medications (SSRIs) during pregnancy. RESULTS: The incidence of congenital anomalies in this study was 1.4%, comparable to general population rates. Rates of low birth weight and preterm births were low, occurring in 2.9% and 6.5% of births, respectively. The low birth weight infants had been exposed to relatively high doses of fluoxetine (40 80 mg/d) throughout pregnancy. Average maternal weight gain in pregnancy was comparable across the three major medication categories (fluoxetine, paroxetine, sertraline). CONCLUSION: After prenatal use of selective serotonin reuptake inhibitor antidepressant medications, neonatal complications and congenital anomalies appear to occur within general population rates. However, maternal use of high doses of fluoxetine throughout pregnancy may be associated with a risk for low birth weight. PMID- 12634663 TI - Placental passage of clarithromycin surpasses other macrolide antibiotics. AB - OBJECTIVE: Infection of the amnion cavity with Ureaplasma urealyticum continues to be a therapeutic challenge. The transplacental transfer rates of macrolide antibiotics are low, and tetracyclines and quinolones are contraindicated in pregnancy. The aim of this study was to investigate placental transfer of clarithromycin in a well-studied placental perfusion model to determine whether clarithromycin surpasses the transfer rate of other macrolide antibiotics in similar models. STUDY DESIGN: Ten placentas that were obtained immediately after delivery were perfused with clarithromycin (3 microg/mL) plus a reference substance (antipyrine). Open circulation placental preparations were used to evaluate steady-state pharmacodynamics and transplacental gradient formation. Drug concentrations were measured by high-performance liquid chromatography. RESULTS: The mean transplacental transfer of clarithromycin was 6.1% (95% CI, 1.8%). CONCLUSION: Because of its enhanced placental passage compared with other macrolide antibiotics, clarithromycin that is given after the first trimester (after embryogenesis) may be an appropriate candidate in treatment trials of genital mycoplasma and ureaplasma infections during pregnancy. PMID- 12634664 TI - Intrapartum fetal heart rate patterns in the prediction of neonatal acidemia. AB - OBJECTIVE: This study was undertaken to correlate changes in the intrapartum electronic fetal heart rate patterns with the development of significant neonatal acidemia. STUDY DESIGN: We identified 488 fetuses at a gestational age of >37 weeks' gestation who had continuous electronic fetal monitoring during labor for the last 2 hours and umbilical artery cord gas analysis performed at delivery. One investigator blinded to the cord gas outcome reviewed all 488 tracings using the National Institute of Child Health and Human Development guidelines for fetal heart rate monitoring. All fetal heart rate tracings with bradycardia were removed from further analysis. The patients were placed in six groups, depending on the absence or presence of normal variability (amplitude >5 beats) during the last hour of monitoring combined with the absence of decelerations or the presence of variable or late decelerations. The relationship between changes in variability and the outcome variables of pH and base deficit in the six groups was assessed with analysis of variance and chi(2) test. Significance was set at the P <.05 level. RESULTS: Patients with normal variability and accelerations, even in the presence of late decelerations or variable decelerations, maintained an umbilical artery pH 7.0 or greater in more than 97% of cases. In the presence of minimal/absent variability (amplitude <5) for at least an hour, the incidence of significant acidemia (pH <7.0) ranged from (12%-31%). CONCLUSION: The most significant intrapartum fetal heart rate parameter to predict the development of significant acidemia is the presence of minimal/absent variability for at least 1 hour as a solitary abnormal finding or in conjunction with late decelerations in the absence of accelerations. Urgent delivery should be considered in these cases after appropriate ancillary testing. PMID- 12634665 TI - The effect of birth weight on vaginal birth after cesarean delivery success rates. AB - OBJECTIVE: The purpose of this study was to evaluate the effect of increasing birth weight on the success rates for a trial of labor in women with one previous cesarean delivery. STUDY DESIGN: To evaluate the effect of increasing birth weight for women who undergo a trial of labor, the medical records of women who had attempted a vaginal birth after cesarean delivery (VBAC) from 1995 through 1999 in 16 community and university hospitals were reviewed retrospectively by trained abstractors. Information was collected about demographics, medical history, obstetric history, neonatal birth weight, complications, treatment, and outcome of the index pregnancy. The analysis was limited to women with singleton gestations with a history of 1 previous cesarean delivery. Because women with previous vaginal deliveries have higher vaginal birth after cesarean delivery success rates, the women were divided into four risk groups on the basis of their birth history. Groups were defined as women with no previous vaginal deliveries (group 1), women with a history of a previous vaginal birth after cesarean delivery (group 2), women with a history of a vaginal delivery before their cesarean delivery (group 3), and a group of women with a vaginal delivery both before and after the previous cesarean delivery (group 4). RESULTS: There were 9960 women with a singleton gestation and a history of one previous cesarean delivery. The overall vaginal birth after cesarean delivery success rate for the cohort was 74%. The overall vaginal birth after cesarean delivery success rates for groups 1, 2, 3, and 4 were 65%, 94%, 83%, and 93%, respectively. An analysis of neonatal birth weights of <4000 g, 4000 to 4249 g, 4250 to 4500 g, and >4500 g in group 1 showed a reduction in vaginal birth after cesarean delivery success rates from 68%, 52%, 45%, and 38%, respectively. In the remaining groups, there was no success rate below 63% for any of the birth weight strata. For group 1, vaginal birth after cesarean delivery success rates were decreased when the indication for the previous cesarean delivery was cephalopelvic disproportion or failure to progress or when the treatment was either an induction or augmentation of labor. The uterine rupture rate was higher in women for group 1 with birth weights of > or =4000 g (relative risk, 2.3; P <.001). CONCLUSION: Women with macrosomic fetuses and a history of a previous vaginal delivery should be informed of the favorable vaginal birth after cesarean delivery success rates. Given the risks of vaginal birth after cesarean delivery, those women with no history of a vaginal delivery should be counseled that the success rates may be <50% when the neonatal birth weight exceeds 4000 g and that the success rates may be even lower if the indication for the previous cesarean delivery was cephalopelvic disproportion or failure to progress or if the treatment requires either induction or augmentation of labor. The uterine rupture rate was 3.6% in women for group 1 with a birth weight > or =4000 g. PMID- 12634666 TI - Early pregnancy threshold vaginal pH and Gram stain scores predictive of subsequent preterm birth in asymptomatic women. AB - OBJECTIVE: The study was undertaken to identify early pregnancy vaginal markers predictive of subsequent preterm birth. STUDY DESIGN: In a multicenter Bacterial Vaginosis (BV) Trial, 21,554 women were screened with a vaginal pH and of these, two populations were studied. These included 12,041 who had a pregnancy outcome in the database and 6838 women who had a vaginal pH of 4.5 or greater and a Gram stain score and a pregnancy outcome in the database. ColorpHast Indicator Strips were used to determine the vaginal pH and the Nugent criteria were used to determine a vaginal Gram stain score of 0 to 10. RESULTS: Delivery at <37, <35, or <32 weeks' gestation was similar for women with a vaginal pH of less than 4.4 or 4.7 (P not significant) but was increased in women with a pH of 5.0 (P =.04,.02,.03, respectively) or with a pH of 5.0 or greater (at each gestational age P <.0001). The effect of pH of 5.0 or greater was similar for women who had a spontaneous preterm birth at each gestational age (P <.0001) or birth weight of less than 2500 g or less than 1500 g (P <.0005). Women with a vaginal pH of 4.5 or greater and a Gram stain score of 9 to 10 (compared with 0-8) had increased preterm births at <37, <35, and <32 weeks' gestation (P <.01), and birth weights less than 2500 g (P <.0001) or less than 1500 g (P <.01). Women whose vaginal pH was 5.0 or greater had a higher prevalence of vaginal fetal fibronectin > or =50 ng/mL (P <.0001), but the proportion of women with a vaginal fetal fibronectin > or =50 mg/mL did not differ by Gram stain score. CONCLUSION: Women with a vaginal pH of 5.0 or greater or a vaginal pH of 4.5 or greater and a Gram stain score of 9 to 10 had significantly increased preterm births at <37, <35, and 32 weeks' gestation and/or a birth weight less than 2500 g or less than 1500 g. PMID- 12634667 TI - A double-blind, randomized, placebo-controlled trial of acyclovir in late pregnancy for the reduction of herpes simplex virus shedding and cesarean delivery. AB - OBJECTIVE: The purpose of this study was to assess the efficacy of acyclovir in the reduction of herpes simplex virus culture and polymerase chain reaction positivity and cesarean delivery. STUDY DESIGN: Women with recurrent genital herpes simplex virus were randomized to acyclovir 400 mg three times daily or placebo from 36 weeks of gestation until delivery. A subset of daily specimens for herpes simplex virus culture and DNA polymerase chain reaction was self collected. Analyses used chi(2), Fisher exact, and Mann-Whitney U tests. RESULTS: Lesions occurred at delivery among 11 of 78 women (14%) who received placebo and 4 of 84 women (5%) who received acyclovir (P =.08). Herpes simplex virus culture and polymerase chain reaction positivity near delivery occurred in 7% and 34% women in the placebo group and 0 and 2% in the acyclovir group (P =.03 and <.01, respectively). Cesarean delivery for herpes simplex virus occurred in 8 of the women (10%) in the placebo group and in 3 of the women (4%) in the acyclovir group (P =.17). Despite reductions in herpes simplex virus detection, 6% of the women who received acyclovir had herpes simplex virus detected by polymerase chain reaction on >20% of days. Neonatal outcomes were similar between groups. CONCLUSION: Acyclovir significantly reduced, but did not eliminate, herpes simplex virus lesions and detection in late pregnancy. PMID- 12634668 TI - Increased polymorphonuclear infiltration and iatrogenic amniotic band after closure of fetoscopic access sites with a bioactive membrane in the rabbit at midgestation. AB - OBJECTIVE: This study was undertaken to evaluate the efficacy and safety of closing the fetoscopy access site in a midgestational rabbit model by using a commercially available bioactive membrane. STUDY DESIGN: Fetoscopy was performed in a total of 100 gestational sacs in 20 does at midgestation (23 days, term = 31 days). In 50 cases (group 1), the fetoscopic access port was closed with a 5-mm patch of biocompatible matrix derived from porcine small intestine containing growth factors (transforming growth factor-beta and fibroblast growth factor beta). Fifty sacs served as positive controls (group 2) and 55 unoperated fetuses were used as negative controls (group 3). At 30 days of gestation, a second-look laparotomy was performed. Outcome parameters were fetal weight, fetal lung weight, fetal lung-to-body weight ratio, and microscopy of the plugging site. RESULTS: Membrane integrity after fetoscopy was restored in 28 of the 40 (70%) of cases in group 1 versus 13 of the 32 (41%) in group 2 (P =.012). Birth weights were comparable (group 1: 30.65 +/- 5.68 g; group 2: 29.70 +/- 5.05 g; group 3: 29.52 +/- 6.25 g; NS), but fetal lung weight (group 1: 0.964 +/- 0.20 g; group 2: 0.798 +/- 0.17 g; P <.01) and fetal lung-to-body weight ratio (group 1: 0.032 +/- 0.0067; group 2: 0.027 +/- 0.0082; P <.05) were significantly higher in the study group. In group 1, cellular proliferation was significantly increased. Polymorphonuclear infiltration was observed in 19 of the 40 (48%) cases in group 1 versus 5 of the 32 (16%) cases in group 2 (P <.05). In one treated sac, a fibrous band joining the two fetal legs without constriction was present. CONCLUSION: The use of a bioactive membrane improved fetal membrane repair rates and decreased incidence of pulmonary hypoplasia in the rabbit but increased polymorphonuclear infiltration. In one amniotic sac, a situation comparable to amniotic band syndrome was documented. PMID- 12634669 TI - Medical education as a science: the quality of evidence for computer-assisted instruction. AB - OBJECTIVE: A marked increase in the number of computer programs for computer assisted instruction in the medical sciences has occurred over the past 10 years. The quality of both the programs and the literature that describe these programs has varied considerably. The purposes of this study were to evaluate the published literature that described computer-assisted instruction in medical education and to assess the quality of evidence for its implementation, with particular emphasis on obstetrics and gynecology. STUDY DESIGN: Reports published between 1988 and 2000 on computer-assisted instruction in medical education were identified through a search of MEDLINE and Educational Resource Identification Center and a review of the bibliographies of the articles that were identified. Studies were selected if they included a description of computer-assisted instruction in medical education, regardless of the type of computer program. Data were extracted with a content analysis of 210 reports. The reports were categorized according to study design (comparative, prospective, descriptive, review, or editorial), type of computer-assisted instruction, medical specialty, and measures of effectiveness. RESULTS: Computer-assisted instruction programs included online technologies, CD-ROMs, video laser disks, multimedia work stations, virtual reality, and simulation testing. Studies were identified in all medical specialties, with a preponderance in internal medicine, general surgery, radiology, obstetrics and gynecology, pediatrics, and pathology. Ninety-six percent of the articles described a favorable impact of computer-assisted instruction in medical education, regardless of the quality of the evidence. Of the 210 reports that were identified, 60% were noncomparative, descriptive reports of new techniques in computer-assisted instruction, and 15% and 14% were reviews and editorials, respectively, of existing technology. Eleven percent of studies were comparative and included some form of assessment of the effectiveness of the computer program. These assessments included pre- and posttesting and questionnaires to score program quality, perceptions of the medical students and/or residents regarding the program, and impact on learning. In one half of these comparative studies, computer-assisted instruction was compared with traditional modes of teaching, such as text and lectures. Six studies compared performance before and after the computer-assisted instruction. Improvements were shown in 5 of the studies. In the remainder of the studies, computer-assisted instruction appeared to result in similar test performance. Despite study design or outcome, most articles described enthusiastic endorsement of the programs by the participants, including medical students, residents, and practicing physicians. Only 1 study included cost analysis. Thirteen of the articles were in obstetrics and gynecology. CONCLUSION: Computer-assisted instruction has assumed to have an increasing role in medical education. In spite of enthusiastic endorsement and continued improvements in software, few studies of good design clearly demonstrate improvement in medical education over traditional modalities. There are no comparative studies in obstetrics and gynecology that demonstrate a clear-cut advantage. Future studies of computer assisted instruction that include comparisons and cost assessments to gauge their effectiveness over traditional methods may better define their precise role. PMID- 12634670 TI - Achieving equipoise in maternal-fetal surgery. PMID- 12634672 TI - Can cervicography be improved? PMID- 12634674 TI - Antenatal steroids in twins. PMID- 12634676 TI - The endometrial-myometrial interface. PMID- 12634678 TI - Complete brachial plexus impairment: a traction-related injury. PMID- 12634679 TI - Interstitial laser coagulation for uterine myomas. PMID- 12634682 TI - Cerebral perfusion pressure, and not cerebral blood flow, may be the critical determinant of intracranial injury in preeclampsia: a new hypothesis. PMID- 12634684 TI - Subcutaneous stitch closure versus subcutaneous drain to prevent wound disruption after cesarean delivery: a randomized clinical trial. PMID- 12634722 TI - The whys and hows of hematopoietic progenitor and stem cell mobilization. AB - Intentional mobilization of hematopoietic/stem cells into the circulation has improved the efficiency of their collection. Transplantation of mobilized blood stem cells to patients with marrow aplasia results in a faster pace of hematopoietic recovery than transplantation of marrow-derived stem cells. Autologous and allogeneic hematopoietic stem cell transplantation are increasingly performed with blood-derived cells. Donors of both autologous and allogeneic blood stem cells do not always respond well to therapies designed to produce mobilization. Autologous donors may respond poorly as a result of myelotoxic damage inflicted by prior antitumor therapy, but this explanation is not valid for allogeneic donors. The mechanism(s) involved in the process of mobilization are incompletely understood. Until these mechanisms are elucidated, methods to improve mobilization vigor on a rational basis will not be obvious. In the meanwhile, clinical observations may provide some hints regarding the whys and hows of mobilization and permit incremental improvements in this process. PMID- 12634723 TI - Total energy expenditure and body composition changes following peripheral blood stem cell transplantation and participation in an exercise programme. AB - The purpose of this investigation was to assess changes in total energy expenditure (TEE), body weight (BW) and body composition following a peripheral blood stem cell transplant and following participation in a 3-month duration, moderate-intensity, mixed-type exercise programme. The doubly labelled and singly labelled water methods were used to measure TEE and total body water (TBW). Body weight and TBW were then used to calculate percentage body fat (%BF), and fat and fat-free mass (FFM). TEE and body composition measures were assessed pretransplant (PI), immediately post-transplant (PII) and 3 months post-PII (PIII). Following PII, 12 patients were divided equally into a control group (CG) or exercise intervention group (EG). While there was no change in TEE between pre and post-transplant, BW (P<0.01) and FFM (P<0.05) significantly decreased during the same period. Participation in the exercise programme led to increases in TEE to levels that were both higher than pre- and post-transplant measures (P<0.01). By PIII, the exercising patients also showed gains in FFM (P<0.01) in association with a reduction in %BF (P<0.05). Exercise has a functionally important role in preserving and increasing skeletal mass in the rehabilitation phase of cancer patients. PMID- 12634724 TI - Reduced intensity conditioning and prophylactic DLI can cure patients with high risk acute leukaemias if complete donor chimerism can be achieved. AB - 23 patients with ALL (n=9) and AML (n=14) underwent nonmyeloablative stem cell transplantation (NST) from an HLA-identical donor after conditioning with fludarabine (180 mg/m(2)), busulfan (8 mg/kg) and anti-T-lymphocyte globulin (40 mg/kg). After NST, 20/23 patients engrafted. Ten out of 14 patients with uncontrolled disease reached complete remission. A multiplex-PCR using short tandem repeats was used for chimerism analysis and detected mixed chimerism (MC) in 14/22 evaluable patients (64%) after NST. Prophylactic donor lymphocyte infusions (DLI) were given to 11/14 patients with MC; MC converted to complete donor chimerism (CC) in 6/11 patients within 2-6 weeks. All patients with persistent MC with or without DLI relapsed during further follow-up. MC predicted impending relapse 4-52 weeks before clinical diagnosis. Ten of 23 patients (43%) are alive 2-34 months after stem cell transplantation. 12 of 23 patients (52%), have died from leukaemia after NST. One out of 23 patients has died from severe sepsis. In conclusion, NST leads to stable engraftment and complete remission in patients with advanced acute leukaemias. NST can cure a substantial proportion of these patients, but the relapse rate is still high. Repeated chimerism analysis is a useful tool to detect recipient cells, especially in patients without molecular markers of disease and can be used to monitor immunomodulatory therapies. MC is unstable in these patients and predicts impending relapse. Prophylactic DLI can convert MC to CC, which seemed to lower relapse risk. PMID- 12634725 TI - Low-dose or intermediate-dose cyclophosphamide plus granulocyte colony stimulating factor for progenitor cell mobilisation in patients with multiple myeloma. AB - Cyclophosphamide (CY) combined with granulocyte colony-stimulating factor (G-CSF) is commonly used to mobilise blood progenitor cells to support high-dose therapy in patients with multiple myeloma (MM). The optimal dose of CY in this setting is unknown. We have retrospectively analysed mobilisation efficiency and need for supportive care in 57 patients with newly diagnosed myeloma previously treated with VAD+/-local radiotherapy. The patients were mobilised either with low-dose CY (LD-CY, 1.2-2 g/m(2)) (n=25) or intermediate-dose CY (ID-CY, 4 g/m(2)) (n=32) plus G-CSF. Both regimens proved to be effective in the progenitor cell mobilisation. At least 2 x 10(6)/kg CD34+ cells were collected from 88% and 84% of the patients with a single apheresis, respectively. Only one patient in the LD CY group (4%) failed to mobilise vs none in the ID-CY group. Patients mobilised with LD-CY plus G-CSF had less toxicity: fewer hospital days during the mobilisation and apheresis procedures (5 vs 9 days, P<0.001), lower frequency of fever (20 vs 73%, P<0.001) and less need for supportive care including platelet transfusions (0 vs 24%, P=0.004) and days on parenteral antibiotics (0 vs 4 days, P<0.001). While these regimens seem to be equally effective in terms of progenitor cell mobilisation in newly diagnosed patients with MM, LD-CY+G-CSF is preferential because of more optimal resource utilisation and more favourable toxicity profile. PMID- 12634726 TI - Osteosarcoma after allogeneic bone marrow transplantation. A report of four cases from the Cooperative Osteosarcoma Study Group (COSS). AB - Osteosarcoma, one of the most frequent secondary malignancies after the treatment of young patients with cancer, has only very rarely been observed in association with hematopoietic stem cell transplantation (HSCT). We report four patients who were identified by searching the database of the Cooperative Osteosarcoma Study Group (COSS) for patients whose osteosarcoma arose following HSCT. Transplant indications had been acute lymphoblastic leukemia (3). and sickle cell disease (1). and the stem cell source was bone marrow in all cases (three allogeneic, one syngeneic). All four had received chemotherapy with alkylators as part of their conditioning regimen and/or first line therapy. The conditioning regimen included total body irradiation in three patients. The osteosarcomas arose at the age (adolescence) and sites (around the knee) typical for the disease. All four patients received chemotherapy as part of multimodal osteosarcoma treatment, and all four are currently alive, three in continuous remission at 5 7/12, 2 11/12, and 0 6/12 years and one with relapsed osteosarcoma at 4 1/12 years. One of the osteosarcoma-free survivors suffered a third malignancy, myelodysplastic syndrome. Osteosarcoma should be included among the secondary malignancies that can arise following HSCT. Multi-modal therapy according to guidelines for de novo osteosarcoma can lead to long-term survival in selected patients. PMID- 12634727 TI - Delivery of Flt3 ligand (Flt3L) using a poloxamer-based formulation increases biological activity in mice. AB - Fms-like tyrosine kinase (Flt3L) is a potent stimulator of hematopoietic progenitor cell (HPC) expansion and mobilization; however, this requires 7-10 days of administration. We investigated whether sustained delivery of Flt3L using a poloxamer-based matrix (PG) could accelerate and/or improve the hematopoietic activity of Flt3L in mice. A single injection of PG-Flt3L stimulated significantly more rapid and greater HPC mobilization to the spleen and peripheral blood than the daily injection of Flt3L formulated in saline. Pharmacokinetic analysis demonstrated that the formulation of Flt3L in PG prolonged its elimination (Tbeta) half-life (2.3-fold) and increased its bioavailability (>two fold) and the time to maximum serum concentration (T(max)) (2.7-fold). Further, coadministration of G-CSF and PG-Flt3L allowed lower doses of Flt3L to be active, with significantly greater hematopoietic and mobilization activity, compared to the same total dose of G-CSF, Flt3L or G-CSF and Flt3L formulated in saline. These data demonstrate that formulation of Flt3L in PG significantly accelerates and increases HPC expansion and mobilization. The observation of increased bioactivity by PG-Flt3L in rodents suggests the potential for improved clinical efficacy of Flt3L by reducing the time required for HPC mobilization. PMID- 12634728 TI - Successful mobilization of peripheral blood stem cells after addition of ancestim (stem cell factor) in patients who had failed a prior mobilization with filgrastim (granulocyte colony-stimulating factor) alone or with chemotherapy plus filgrastim. AB - This study assessed the ability of recombinant human stem cell factor (rHuSCF) to mobilize stem cells in 44 patients who had failed a prior mobilization (CD34(+) yield 0.5-1.9 x 10(6)/kg BW) with filgrastim-alone or chemotherapy-plus filgrastim. The same mobilization regimen was used with the addition of rHuSCF. In the filgrastim-alone group (n=13), rHuSCF 20 microg/kg was started 3 days before filgrastim and continued for the duration of filgrastim. In the chemotherapy-plus-filgrastim group (n=31), rHuSCF 20 microg/kg/day plus filgrastim 5-10 microg/kg/day were administered concurrently. Leukaphereses were continued to a maximum of four procedures or a target of >or=3 x 10(6) CD34(+) cells/kg. In both groups, CD34(+) yield (x 10(6)/kg BW) of the study mobilization was higher than that of the prior mobilization (median: 2.42 vs 0.84 P=0.002 and 1.64 vs 0.99 P=<0.001, respectively). In all 54 and 45% of patients in the filgrastim-alone group and chemotherapy-plus-filgrastim group, respectively, reached the threshold yield of 2 x 10(6)/kg. The probability of a successful mobilization was the same in those with a CD34+ yield of 0.5-0.75 x 10(6)/kg BW in the prior mobilization as in those with 0.76-1.99 x 10(6)/kg BW. Downmodulation of c-kit expression and a lower percentage of Thy-1 positivity in the mobilized CD34(+) cells were noted in the successful mobilizers compared with those in the poor mobilizers. This study shows that rhuSCF is effective in approximately half the patients who had failed a prior mobilization and allows them to proceed to transplant. It also points to the likely role of the SCF/c-kit ligand pair in mobilization. PMID- 12634729 TI - Polyoma viruria following T-cell-depleted allogeneic transplants using Campath 1H: incidence and outcome in relation to graft manipulation, donor type and conditioning. AB - Haemorrhagic cystitis (HC) is an important cause of morbidity following stem cell transplantation (SCT) and has been associated with polyoma virus infection. We studied the incidence and outcome of polyoma virus infection in 58 T-cell depleted SCT patients. T-cell depletion was carried out using Campath-1H, either 10 or 20 mg in vitro (n=33) or 50 or 100 mg in vivo (n=25) following conventional (n=35) or nonmyeloablative conditioning (n=23). A total of 21 patients (36%) had polyoma viruria at a median of 35 days (5-114); 30% among patients receiving Campath in vitro and 44% among those given in vivo. The only risk factor for polyoma viruria was graft-versus-host disease GVHD grade >or=2. The onset of polyoma viruria coincided with Cytomegalovirus (CMV) reactivation in all six patients who reactivated both viruses. Prolonged viruria (defined as polyoma viruria >2 weeks) was documented in 10 patients (17%) and this was associated with GVHD >or=grade 2. HC occurred in four patients. Prolonged viruria was associated with HC only in patients receiving unrelated donor grafts following conventional conditioning. HC was not observed following nonmyeloablative conditioning despite a higher incidence of prolonged viruria. Thus, HC was uncommon in patients with polyoma viruria following T-cell depletion with Campath, particularly after reduced intensity conditioning. PMID- 12634730 TI - Chimerism analysis following allogeneic peripheral blood stem cell transplantation with reduced-intensity conditioning. AB - We have performed a prospective study to evaluate early chimerism and its kinetics after allogeneic peripheral blood stem cell transplantation among 68 patients who received a reduced-intensity conditioning (RIC) regimen with fludarabine plus melphalan (n=40) or busulphan (n=28). Chimerism was analyzed by polymerase chain reaction amplification of short tandem repeats in unfractionated (UF) and/or fractionated nucleated cells from bone marrow and peripheral blood (PB). All of the patients showed initial donor engraftment and no patient presented primary or secondary graft failure. In UF samples, the probability of achieving stable complete donor chimerism (CDC) in PB within the first 6 months was 70% on day +30, 85% on day +100 and 95% on day +180. CDC in granulocytes was observed in nearly all cases from day +30 onwards. CDC in T cells, however, differed among melphalan and busulphan recipients during the first 3 months (100 vs 0% on day +30 and 93 vs 20% on day +90, respectively). In multivariate analysis, the only significant variable associated with the achievement of early CDC was having received more than two lines of chemotherapy pretransplant (P<0.02). No correlation was found between the rate of achieving early CDC and the occurrence of acute graft-versus-host disease (GVHD) or disease progression post-transplant. In multivariate analysis, the only variable that influenced the incidence of disease progression post-transplant was the development of chronic extensive GVHD (P<0.05). In conclusion, a state of CDC is readily obtained within the first 6 months after our RIC protocols. Donor myeloid engraftment occurs rapidly in all cases, while early T-cell CDC is more common in more immunosuppressed hosts and, perhaps, in melphalan recipients. PMID- 12634731 TI - Engraftment syndrome following autologous hematopoietic stem cell transplantation: definition of diagnostic criteria. AB - Engraftment syndrome (ES) is an increasingly reported complication of hematopoietic stem cell transplantation (HSCT). In order to better characterize the clinical criteria for the diagnosis of ES, we retrospectively analyzed 125 autologous HSCT recipients. ES was first defined as the presence of noninfectious fever plus skin rash. Patients with and without these findings were compared (univariate and multivariate analyses) regarding the presence of weight gain, hypoalbuminemia, pulmonary infiltrates, diarrhea, neurological manifestations and jaundice. The variables that are significantly more frequent in patients with fever and skin rash were incorporated in the definition criteria. The final diagnostic criteria were noninfectious fever plus any of the following: skin rash, pulmonary infiltrates or diarrhea. The incidence of ES was 20%. The single risk factor for ES by multivariate analysis was a diagnosis other than Hodgkin's disease (odds ratio 6.17, 95% confidence interval 1.38-27.78). Patients with ES received empirical antifungal therapy more frequently than patients without the syndrome (40 vs 19%, P=0.03), and had a longer duration of hospitalization (P=0.0007). The prospective application of these diagnostic criteria may have a favorable impact on the early diagnosis of the syndrome, with the initiation of corticosteroids and a reduction in the unnecessary use of antimicrobial agents. PMID- 12634732 TI - Refractory Evans' syndrome treated with allogeneic SCT followed by DLI. Demonstration of a graft-versus-autoimmunity effect. AB - Evans' syndrome, a combination of autoimmune haemolytic anaemia and autoimmune (idiopathic) thrombocytopenic purpura, is generally harder to treat and more refractory than the single entities. In a male patient with refractory disease, predominantly thrombocytopenic, an allogeneic reduced intensity BMT from his human leukocyte antigen (HLA)-identical sister was followed by a dramatic platelet peak while he was still experiencing initial engraftment (presumably of autologous origin), but subsequently by progressive relapse associated with mixed chimerism. Five gradually incremental DLI achieved complete donor chimerism, which was associated not only with grade II graft-versus-host disease (GVHD), but also with complete clinical and biological remission for 2 years post-transplant. Long-term FU is necessary before claiming that allogeneic stem cell transplantation (SCT) is capable of curing an autoimmune blood disease. However, there is evidence for a graft-versus-autoimmunity effect in this case. PMID- 12634733 TI - Successful umbilical cord blood stem cell transplantation for chronic granulomatous disease. AB - Chronic granulomatous disease (CGD) causes growth failure, inflammatory lung damage and often early death. Prophylactic cotrimoxazole improves medium-term survival, but cannot prevent inflammatory sequelae. We report the first patient with CGD who underwent successful HLA identical sibling umbilical cord stem cell transplantation (UCSCT) after myeloablative conditioning. The patient presented with colitis, confirmed as CGD at 2 years of age. Following BU16/CY200 conditioning, he had UCSCT from his unaffected HLA identical sister. A year post transplant, his colitis had resolved clinically and on radioisotope scan growth has improved. Neutrophil oxidative burst was 92% normal with full donor lymphocyte reconstitution. PMID- 12634734 TI - Nonmyeloablative allogeneic hematopoietic stem cell transplantation for treatment of Dyskeratosis congenita. AB - We describe the treatment of a 10-year-old girl with autosomal recessive Dyskeratosis congenita (DC), neutropenia, thrombocytopenia and combined immunodeficiency by nonmyeloablative hematopoietic stem cell transplantation. The conditioning regimen consisted of fludarabine 30 mg/m(2)/day (days -5, -4, -3) and 2 Gy TBI (0.07 Gy/min; day 0). For graft-versus-host disease (GVHD) prophylaxis a course of intravenous MMF and CSA was administered. At 2 years after transplantation of granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood stem cells from a healthy 11-year-old HLA-identical brother, peripheral blood counts and T- and B-cell functions have completely normalized and donor chimerism was 100% in all cell lineages. No GVHD occurred. Neurological examination and lung function remained normal. The current transplantation regimen appears suitable, safe and efficacious in patients with DC. PMID- 12634735 TI - Successful treatment of disseminated fusariosis. PMID- 12634738 TI - Time to unite Islam and science. PMID- 12634736 TI - Metastatic gastric adenocarcinoma following allogeneic stem cell transplantation in a patient with acute myelogenous leukemia. PMID- 12634739 TI - A little protectionism goes a long way. PMID- 12634740 TI - Academies wrestle with issue of Islam's flagging science base. PMID- 12634741 TI - Bush seeks to beef up US nuclear-weapons arsenal. PMID- 12634743 TI - Auction of DNA archive cancelled. PMID- 12634742 TI - Biotech project in turmoil as Michigan balances books. PMID- 12634744 TI - Tougher rules aim to prevent gene flow into crops. PMID- 12634745 TI - Senator rebuffs academy over Arctic oil. PMID- 12634747 TI - Russia pulls out of Antarctic station. PMID- 12634746 TI - Hydrogen cars fuel debate on basic research. PMID- 12634748 TI - Max Planck plans double blow to chemistry. PMID- 12634749 TI - Danish biotech centre faces axe. PMID- 12634751 TI - Cosmology gets real. PMID- 12634752 TI - Chinese agribiotech: Against the grain. PMID- 12634753 TI - Universities could gain from backing biotech. PMID- 12634754 TI - Work on 'non-lethal' weapons should be limited too. PMID- 12634755 TI - Constellations in a cellular universe. PMID- 12634761 TI - Hormone evolution: The key to signalling. PMID- 12634762 TI - Developmental biology: Flowers' wings, fruitflies' petals. PMID- 12634763 TI - Astronomy: Atmosphere out of that world. PMID- 12634764 TI - Developmental biology: Guidance molecule goes global. PMID- 12634765 TI - Behavioural science: Fair's fair. PMID- 12634766 TI - Planetary science: The core of planet formation. PMID- 12634769 TI - Surface science: View from the edge. PMID- 12634768 TI - Evolutionary biology: Teeth as tools. PMID- 12634770 TI - Solar System: Close encounter of the cometary kind. PMID- 12634771 TI - Medicine: Smoke signals for lung disease. PMID- 12634773 TI - Palaeontology: Human footprints in Pleistocene volcanic ash. PMID- 12634774 TI - Atmospheric science: Ultraviolet light and leaf emission of NO(x). PMID- 12634775 TI - Conservation: Reproductive collapse in saiga antelope harems. PMID- 12634776 TI - Earth science: Mantle deformation or processing artefact? PMID- 12634779 TI - Nanosecond radio bursts from strong plasma turbulence in the Crab pulsar. AB - The Crab pulsar was discovered by the occasional exceptionally bright radio pulses it emits, subsequently dubbed 'giant' pulses. Only two other pulsars are known to emit giant pulses. There is no satisfactory explanation for the occurrence of giant pulses, nor is there a complete theory of the pulsar emission mechanism in general. Competing models for the radio emission mechanism can be distinguished by the temporal structure of their coherent emission. Here we report the discovery of isolated, highly polarized, two-nanosecond subpulses within the giant radio pulses from the Crab pulsar. The plasma structures responsible for these emissions must be smaller than one metre in size, making them by far the smallest objects ever detected and resolved outside the Solar System, and the brightest transient radio sources in the sky. Only one of the current models--the collapse of plasma-turbulent wave packets in the pulsar magnetosphere--can account for the nanopulses we observe. PMID- 12634778 TI - Detrimental effects of sanctions on human altruism. AB - The existence of cooperation and social order among genetically unrelated individuals is a fundamental problem in the behavioural sciences. The prevailing approaches in biology and economics view cooperation exclusively as self interested behaviour--unrelated individuals cooperate only if they face economic rewards or sanctions rendering cooperation a self-interested choice. Whether economic incentives are perceived as just or legitimate does not matter in these theories. Fairness-based altruism is, however, a powerful source of human cooperation. Here we show experimentally that the prevailing self-interest approach has serious shortcomings because it overlooks negative effects of sanctions on human altruism. Sanctions revealing selfish or greedy intentions destroy altruistic cooperation almost completely, whereas sanctions perceived as fair leave altruism intact. These findings challenge proximate and ultimate theories of human cooperation that neglect the distinction between fair and unfair sanctions, and they are probably relevant in all domains in which voluntary compliance matters--in relations between spouses, in the education of children, in business relations and organizations as well as in markets. PMID- 12634780 TI - An extended upper atmosphere around the extrasolar planet HD209458b. AB - The planet in the system HD209458 is the first one for which repeated transits across the stellar disk have been observed. Together with radial velocity measurements, this has led to a determination of the planet's radius and mass, confirming it to be a gas giant. But despite numerous searches for an atmospheric signature, only the dense lower atmosphere of HD209458b has been observed, through the detection of neutral sodium absorption. Here we report the detection of atomic hydrogen absorption in the stellar Lyman alpha line during three transits of HD209458b. An absorption of 15 +/- 4% (1sigma) is observed. Comparison with models shows that this absorption should take place beyond the Roche limit and therefore can be understood in terms of escaping hydrogen atoms. PMID- 12634781 TI - Observation of two-dimensional discrete solitons in optically induced nonlinear photonic lattices. AB - Nonlinear periodic lattices occur in a large variety of systems, such as biological molecules, nonlinear optical waveguides, solid-state systems and Bose Einstein condensates. The underlying dynamics in these systems is dominated by the interplay between tunnelling between adjacent potential wells and nonlinearity. A balance between these two effects can result in a self-localized state: a lattice or 'discrete' soliton. Direct observation of lattice solitons has so far been limited to one-dimensional systems, namely in arrays of nonlinear optical waveguides. However, many fundamental features are expected to occur in higher dimensions, such as vortex lattice solitons, bright lattice solitons that carry angular momentum, and three-dimensional collisions between lattice solitons. Here, we report the experimental observation of two-dimensional (2D) lattice solitons. We use optical induction, the interference of two or more plane waves in a photosensitive material, to create a 2D photonic lattice in which the solitons form. Our results pave the way for the realization of a variety of nonlinear localization phenomena in photonic lattices and crystals. Finally, our observation directly relates to the proposed lattice solitons in Bose-Einstein condensates, which can be observed in optically induced periodic potentials. PMID- 12634783 TI - Core formation in planetesimals triggered by permeable flow. AB - The tungsten isotope composition of meteorites indicates that core formation in planetesimals occurred within a few million years of Solar System formation. But core formation requires a mechanism for segregating metal, and the 'wetting' properties of molten iron alloy in an olivine-rich matrix is thought to preclude segregation by permeable flow unless the silicate itself is partially molten. Excess liquid metal over a percolation threshold, however, can potentially create permeability in a solid matrix, thereby permitting segregation. Here we report the percolation threshold for molten iron-sulphur compounds of approximately 5 vol.% in solid olivine, based on electrical conductivity measurements made in situ at high pressure and temperature. We conclude that heating within planetesimals by decay of short-lived radionuclides can increase temperature sufficiently above the iron-sulphur melting point (approximately 1,000 degrees C) to trigger segregation of iron alloy by permeable flow within the short timeframe indicated by tungsten isotopes. We infer that planetesimals with radii greater than about 30 km and larger planetary embryos are expected to have formed cores very early, and these objects would have contained much of the mass in the terrestrial region of the protoplanetary nebula. The Earth and other terrestrial planets are likely therefore to have formed by accretion of previously differentiated planetesimals, and Earth's core may accordingly be viewed as a blended composite of pre-formed cores. PMID- 12634782 TI - Self-organization of dissolved organic matter to micelle-like microparticles in river water. AB - In aquatic systems, the concept of the 'microbial loop' is invoked to describe the conversion of dissolved organic matter to particulate organic matter by bacteria. This process mediates the transfer of energy and matter from dissolved organic matter to higher trophic levels, and therefore controls (together with primary production) the productivity of aquatic systems. Here we report experiments on laboratory incubations of sterile filtered river water in which we find that up to 25% of the dissolved organic carbon (DOC) aggregates abiotically to particles of diameter 0.4-0.8 micrometres, at rates similar to bacterial growth. Diffusion drives aggregation of low- to high-molecular-mass DOC and further to larger micelle-like microparticles. The chemical composition of these microparticles suggests their potential use as food by planktonic bacterivores. This pathway is apparent from differences in the stable carbon isotope compositions of picoplankton and the microparticles. A large fraction of dissolved organic matter might therefore be channelled through microparticles directly to higher trophic levels--bypassing the microbial loop--suggesting that current concepts of carbon conversion in aquatic systems require revision. PMID- 12634784 TI - Escalation of a coevolutionary arms race through host rejection of brood parasitic young. AB - Cuckoo nestlings that evict all other young from the nest soon after hatching impose a high reproductive cost on their hosts. In defence, hosts have coevolved strategies to prevent brood parasitism. Puzzlingly, they do not extend beyond the egg stage. Thus, hosts adept at recognizing foreign eggs remain vulnerable to exploitation by cuckoo nestlings. Here we show that the breach of host egg defences by cuckoos creates a new stage in the coevolutionary cycle. We found that defences used during the egg-laying period by host superb fairy-wrens (Malurus cyaneus) are easily evaded by the Horsfield's bronze-cuckoo (Chrysococcyx basalis), a specialist fairy-wren brood parasite. However, although hosts never deserted their own broods, they later abandoned 40% of nests containing a lone Horsfield's bronze-cuckoo nestling, and 100% of nests with a lone shining bronze-cuckoo nestling (Chrysococcyx lucidus), an occasional fairy wren brood parasite. Our experiments demonstrate that host discrimination against evictor-cuckoo nestlings is possible, and suggest that it has selected for the evolution of nestling mimicry in bronze-cuckoos. PMID- 12634785 TI - Growth dynamics underlying petal shape and asymmetry. AB - Development commonly involves the generation of complex shapes from simpler ones. One way of following this process is to use landmarks to track the fate of particular points in a developing organ, but this is limited by the time over which it can be monitored. Here we use an alternative method, clonal analysis, whereby dividing cells are genetically marked and their descendants identified visually, to observe the development of Antirrhinum (snapdragon) petals. Clonal analysis has previously been used to estimate growth parameters of leaves and Drosophila wings but these results were not integrated within a dynamic growth model. Here we develop such a model and use it to show that a key aspect of shape -petal asymmetry--in the petal lobe of Antirrhinum depends on the direction of growth rather than regional differences in growth rate. The direction of growth is maintained parallel to the proximodistal axis of the flower, irrespective of changes in shape, implying that long-range signals orient growth along the petal as a whole. Such signals may provide a general mechanism for orienting growth in other growing structures. PMID- 12634786 TI - Selection of evolutionarily conserved mucosal-associated invariant T cells by MR1. AB - The evolutionary conservation of T lymphocyte subsets bearing T-cell receptors (TCRs) using invariant alpha-chains is indicative of unique functions. CD1d restricted natural killer T (NK-T) cells that express an invariant Valpha14 TCRalpha chain have been implicated in microbial and tumour responses, as well as in auto-immunity. Here we show that T cells that express the canonical hValpha7.2 Jalpha33 or mValpha19-Jalpha33 TCR rearrangement are preferentially located in the gut lamina propria of humans and mice, respectively, and are therefore genuine mucosal-associated invariant T (MAIT) cells. Selection and/or expansion of this population requires B lymphocytes, as MAIT cells are absent in B-cell deficient patients and mice. In addition, we show that MAIT cells are selected and/or restricted by MR1, a monomorphic major histocompatibility complex class I related molecule that is markedly conserved in diverse mammalian species. MAIT cells are not present in germ-free mice, indicating that commensal flora is required for their expansion in the gut lamina propria. This indicates that MAIT cells are probably involved in the host response at the site of pathogen entry, and may regulate intestinal B-cell activity. PMID- 12634787 TI - Loss of integrin alpha(v)beta6-mediated TGF-beta activation causes Mmp12 dependent emphysema. AB - Integrins are heterodimeric cell-surface proteins that regulate cell growth, migration and survival. We have shown previously that the epithelial-restricted integrin alpha(v)beta6 has another critical function; that is, it binds and activates latent transforming growth factor-beta (TGF-beta). Through a global analysis of pulmonary gene expression in the lungs of mice lacking this integrin (Itgb6 null mice) we have identified a marked induction of macrophage metalloelastase (Mmp12)--a metalloproteinase that preferentially degrades elastin and has been implicated in the chronic lung disease emphysema. Here we report that Itgb6-null mice develop age-related emphysema that is completely abrogated either by transgenic expression of versions of the beta6 integrin subunit that support TGF-beta activation, or by the loss of Mmp12. Furthermore, we show that the effects of Itgb6 deletion are overcome by simultaneous transgenic expression of active TGF-beta1. We have uncovered a pathway in which the loss of integrin mediated activation of latent TGF-beta causes age-dependent pulmonary emphysema through alterations of macrophage Mmp12 expression. Furthermore, we show that a functional alteration in the TGF-beta activation pathway affects susceptibility to this disease. PMID- 12634788 TI - Functional and spatial segregation of secretory vesicle pools according to vesicle age. AB - Synaptic terminals and neuroendocrine cells are packed with secretory vesicles, only a few of which are docked at the plasma membrane and readily releasable. The remainder are thought to constitute a large cytoplasmic reserve pool awaiting recruitment into the readily releasable pool (RRP) for exocytosis. How vesicles are prioritized in recruitment is still unknown: the choice could be random, or else the oldest or the newest ones might be favoured. Here we show, using a fluorescent cargo protein that changes colour with time, that vesicles in bovine adrenal chromaffin cells segregate into distinct populations, based on age. Newly assembled vesicles are immobile (morphologically docked) at the plasma membrane shortly after biogenesis, whereas older vesicles are mobile and located deeper in the cell. Different secretagogues selectively release vesicles from the RRP or, surprisingly, selectively from the deeper cytoplasmic pool. Thus, far from being equal, vesicles are segregated functionally and spatially according to age. PMID- 12634789 TI - Insight into a natural Diels-Alder reaction from the structure of macrophomate synthase. AB - The Diels-Alder reaction, which forms a six-membered ring from an alkene (dienophile) and a 1,3-diene, is synthetically very useful for construction of cyclic products with high regio- and stereoselectivity under mild conditions. It has been applied to the synthesis of complex pharmaceutical and biologically active compounds. Although evidence on natural Diels-Alderases has been accumulated in the biosynthesis of secondary metabolites, there has been no report on the structural details of the natural Diels-Alderases. The function and catalytic mechanism of the natural Diels-Alderase are of great interest owing to the diversity of molecular skeletons in natural Diels-Alder adducts. Here we present the 1.70 A resolution crystal structure of the natural Diels-Alderase, fungal macrophomate synthase (MPS), in complex with pyruvate. The active site of the enzyme is large and hydrophobic, contributing amino acid residues that can hydrogen-bond to the substrate 2-pyrone. These data provide information on the catalytic mechanism of MPS, and suggest that the reaction proceeds via a large scale structural reorganization of the product. PMID- 12634792 TI - From genomics to proteomics. AB - Proteomics is the study of the function of all expressed proteins. Tremendous progress has been made in the past few years in generating large-scale data sets for protein-protein interactions, organelle composition, protein activity patterns and protein profiles in cancer patients. But further technological improvements, organization of international proteomics projects and open access to results are needed for proteomics to fulfil its potential. PMID- 12634793 TI - Mass spectrometry-based proteomics. AB - Recent successes illustrate the role of mass spectrometry-based proteomics as an indispensable tool for molecular and cellular biology and for the emerging field of systems biology. These include the study of protein-protein interactions via affinity-based isolations on a small and proteome-wide scale, the mapping of numerous organelles, the concurrent description of the malaria parasite genome and proteome, and the generation of quantitative protein profiles from diverse species. The ability of mass spectrometry to identify and, increasingly, to precisely quantify thousands of proteins from complex samples can be expected to impact broadly on biology and medicine. PMID- 12634794 TI - Protein analysis on a proteomic scale. AB - The long-term challenge of proteomics is enormous: to define the identities, quantities, structures and functions of complete complements of proteins, and to characterize how these properties vary in different cellular contexts. One critical step in tackling this goal is the generation of sets of clones that express a representative of each protein of a proteome in a useful format, followed by the analysis of these sets on a genome-wide basis. Such studies enable genetic, biochemical and cell biological technologies to be applied on a systematic level, leading to the assignment of biochemical activities, the construction of protein arrays, the identification of interactions, and the localization of proteins within cellular compartments. PMID- 12634795 TI - From words to literature in structural proteomics. AB - Technical advances on several frontiers have expanded the applicability of existing methods in structural biology and helped close the resolution gaps between them. As a result, we are now poised to integrate structural information gathered at multiple levels of the biological hierarchy - from atoms to cells - into a common framework. The goal is a comprehensive description of the multitude of interactions between molecular entities, which in turn is a prerequisite for the discovery of general structural principles that underlie all cellular processes. PMID- 12634796 TI - Disease proteomics. AB - The sequencing of the human genome and that of numerous pathogens has opened the door for proteomics by providing a sequence-based framework for mining proteomes. As a result, there is intense interest in applying proteomics to foster a better understanding of disease processes, develop new biomarkers for diagnosis and early detection of disease, and accelerate drug development. This interest creates numerous opportunities as well as challenges to meet the needs for high sensitivity and high throughput required for disease-related investigations. PMID- 12634797 TI - Biomedical informatics for proteomics. AB - Success in proteomics depends upon careful study design and high-quality biological samples. Advanced information technologies, and also an ability to use existing knowledge to the full, will be crucial in making sense of the data. Despite its genome-scale potential, proteome analysis is at a much earlier stage of development than genomics and gene expression (microarray) studies. Fundamental issues involving biological variability, pre-analytic factors and analytical reproducibility remain to be resolved. Consequently, the analysis of proteomics data is currently informal and relies heavily on expert opinion. Databases and software tools developed for the analysis of molecular sequences and microarrays are helpful, but are limited owing to the unique attributes of proteomics data and differing research goals. PMID- 12634798 TI - Eastern promises. PMID- 12634799 TI - California dreaming: San Diego. PMID- 12634800 TI - The dog that came in from the cold. PMID- 12634803 TI - The FMR1 CGG repeat and linked microsatellite markers in two Basque valleys. AB - Fragile X syndrome is associated with an unstable CGG repeat sequence in the 5' untranslated region of the first exon of the FMR1 gene. The present study involved the evaluation of factors implicated in CGG repeat stability in a normal sample from two Basque valleys (Markina and Arratia), to discover whether the Basque population shows allelic diversity and to identify factors involved, by using the data in conjunction with previous findings. The study was based on a sample of 204 and 58 X chromosomes from the Markina and Arratia valleys, respectively. The CGG repeat, the AGG interspersion and two flanking microsatellite markers, FRAXAC1 and DXS548, were examined. In the Markina valley, gray zone alleles (> or =35 CGG repeats) were associated with anchoring AGGs, with the longest 3' pure CGG repeats of the valley (=15), with the 5' instability structure 9+n and with one principal fragile X FRAXAC1-DXS548 haplotype 42-50. In the Arratia valley, gray zone alleles (> or =35 CGG repeats) showed the highest frequency among the Basque samples analyzed, and were associated with anchoring AGGs, with the longest 3' pure repeats (> or =20), with the 5' instability structure 9+n and with one "normal" FRAXAC1-DXS548 haplotype 38-40 (these data from Arratia suggest the existence of a "protective" haplotype). The results showed, on the one hand, differences between Markina and Arratia in factors implicated in CGG repeat instability and, on the other hand, a great similarity between the general Basque sample from Biscay and the Markina valley. PMID- 12634804 TI - The Great Wall of China: a physical barrier to gene flow? AB - One population from each of six plant species along both sides of the Juyong-guan Great Wall, together with one population from each of five species along both sides of a path on a mountain top near Juyong-guan, were selected to study the effect of the Great Wall as a barrier on genetic differentiation between two subpopulations using RAPD markers. Significant genetic differentiation was found between the subpopulations on both sides of the Great Wall. A wind-pollinated woody species, Ulmus pumila, showed less genetic differentiation than four insect pollinated species: Prunus armeniaca, Ziziphus jujuba, Vitex negundo, and Heteropappus hispidus. Cleistogenes caespitosa, a wind-pollinated perennial herb, displayed more genetic differentiation between subpopulations than the insect pollinated species because of its propagation strategy. Although AMOVA analysis showed that subpopulations divided by a mountain path had diverged genetically, the variance component between the subpopulations on both sides of the Great Wall was significantly larger than that between the subpopulations at the control site. Therefore, it is reasonable to deduce that the Juyong-guan Great Wall has served as a physical barrier to gene flow between subpopulations separated for more than 600 years. PMID- 12634805 TI - Genetic differentiation in the Agave deserti (Agavaceae) complex of the Sonoran desert. AB - The Agave deserti complex, comprising A. deserti, A. cerulata and A. subsimplex, represents a group of species and subspecies with a near allopatric distribution and clear differences in morphology. Genetic differentiation and taxonomic status with respect to spatial distribution of 14 populations of the complex were analyzed in an effort to understand the evolution and speciation process within the genus. Allelic frequencies, levels of genetic variation, expected heterozygosity (H(S)), proportion of polymorphic loci (P), and genetic differentiation (theta and Nei's genetic distance) were estimated using 41 putative RAPD loci. All three species show high levels of genetic variation (H(S)=0.12-0.29, P=63.4-95.1), and low genetic differentiation between populations and species (theta populations=0.14+/-0.02 (SE); G(st)=0.11+/-0.02). Accordingly, gene flow among populations was estimated as high by three different methods (N(m)=2.91-6.14). Nei's genetic distances between the three species were low compared to the values obtained from other Agavaceae, and there was no clear correlation with taxonomic divisions. In a UPGMA analysis, A. subsimplex and A. cerulata formed exclusive monospecific clusters, whereas the A. deserti populations appear in more than one cluster together with other species. The results were consistent with a pattern of genetic isolation by distance. PMID- 12634806 TI - Mapping quantitative trait loci underlying triploid endosperm traits. AB - Endosperm, which is derived from two polar nuclei fusing with one sperm, is a triploid tissue in cereals. Endosperm tissue determines the grain quality of cereals. Improving grain quality is one of the important breeding objectives in cereals. However, current statistical methods for mapping quantitative trait loci (QTL) under diploid genetic control have not been effective for dealing with endosperm traits because of the complexity of their triploid inheritance. In this paper, we derive for the first time the conditional probabilities of F(3) endosperm QTL genotypes given different flanking marker genotypes in F(2) plants. Using these probabilities, we develop a multiple linear regression method implemented via the iteratively reweighted least-squares (IRWLS) algorithm and a maximum likelihood method (ML) implemented via the expectation-maximization (EM) algorithm to map QTL underlying endosperm traits. We use the mean value of endosperm traits of F(3) seeds as the dependent variable and the expectations of genotypic indicators for additive and dominance effect of a putative QTL flanked by a pair of markers as independent variables for IRWLS mapping. However, if an endosperm trait is measured quantitatively using a single endosperm sample, the ML mapping method can be used to separate the two dominance effects. Efficiency of the methods is verified through extensive Monte Carlo simulation studies. Results of simulation show that the proposed methods provide accurate estimates of both the QTL effects and locations with very high statistical power. With these methods, we are now ready to map endosperm traits, as we can for regular quantitative trait under diploid control. PMID- 12634807 TI - Incomplete sexual isolation in sympatry between subspecies of the butterfly Danaus chrysippus (L.) and the creation of a hybrid zone. AB - Subspecies chrysippus, dorippus and alcippus of the butterfly Danaus chrysippus differ at three biallelic colour gene loci. They have partially vicariant distributions, but their ranges overlap over a substantial part of central and East Africa, where hybridism is commonplace. We now report that the West African subspecies alcippus differs from other subspecies, not only in nuclear genotype but also in mitochondrial haplotype in both allopatry and sympatry. The maintenance of concordant nuclear and cytoplasmic genetic differences in sympatry, and in the face of hybridisation, is prima facie evidence for sexual isolation. Other evidence that suggests alcippus may be isolated from chrysippus and dorippus include differences in sex ratio (SR), heterozygote deficiency at one site and deduced differences in patterns of migration. We suggest that, within the hybrid zone, differential infection of subspecies by a male-killing Spiroplasma bacterium causes SR differences that restrict female choice, triggering rounds of heterotypic mating and consequent heterozygote excess that is largely confined to females. The absence of these phenomena from hybrid populations that test negative for Spiroplasma supports the hypothesis. The incomplete sexual isolation and partial vicariance of alcippus suggests that it is a nascent species. PMID- 12634808 TI - Somatic stability of microsatellite loci in Eastern white pine, Pinus strobus L. AB - Variation at nuclear- and chloroplast-encoded microsatellite loci was studied among and within clonally propagated individuals of Eastern white pine. Total DNA was extracted and assayed from gamete-bearing tissue (megagametophytes) located on six different branch positions on each of 12 individual genets. No within individual variation was observed among 12 loci studied. Estimates of numbers of mitotic cell divisions required to produce the tissue used as the source of genomic DNA were obtained by combining tree growth and anatomical data. This allowed for the calculation of upper bound estimates of numbers of mutations per locus per somatic cell division. The estimated somatic mutation rate was found to be substantially lower than those published for genomic microsatellite mutation rates in other plant species. PMID- 12634809 TI - Genetic diversity and population structure of the Italian fungi belonging to the taxa Pleurotus eryngii (DC.:Fr.) Quel and P. ferulae (DC.:Fr.) Quel. AB - A study using allozymes and PCR fingerprinting was conducted to estimate the genetic diversity of Italian populations of two economically important cultivated fungal taxa, Pleurotus eryngii and P. ferulae. Very little is known about the genetic diversity distribution pattern of these taxa. Heterozygote deficiency was observed at few loci; in fact the inbreeding coefficients were not high, which demonstrates that mechanisms restrain the inbreeding act at the local level. Estimates of genetic differentiation indicated a pattern of greater variation within, rather than between, populations. These results were supported by AMOVA analysis, which attributed a low proportion of the total genetic variation to large geographical scale divergence, and indicated that most of the genetic diversity was because of differences within populations. This distribution pattern of genetic variation of P. eryngii and P. ferulae populations seems to be the result of high gene flow, by efficient basidiospore dispersal, and outcrossing mechanisms, which restrain inbreeding within populations. PMID- 12634810 TI - Gene flow between arrhenotokous and thelytokous populations of Venturia canescens (Hymenoptera). AB - In the solitary parasitoid wasp Venturia canescens both arrhenotokously (sexual) and thelytokously (parthenogenetical) reproducing individuals occur sympatrically. We found in the laboratory that thelytokous wasps are able to mate, receive and use sperm of arrhenotokous males. Using nuclear (amplified fragment length polymorphism, virus-like protein) and mitochondrial (restriction fragment length polymorphism) markers, we show the occurrence of gene flow from the arrhenotokous to the thelytokous mode in the field. Our results reinforce the paradox of sex in this species. PMID- 12634811 TI - Fine-scale genetic structure and gene flow within Costa Rican populations of mahogany (Swietenia macrophylla). AB - Fine-scale structure of genetic diversity and gene flow were analysed in three Costa Rican populations of mahogany, Swietenia macrophylla. Population differentiation estimated using AFLPs and SSRs was low (38.3 and 24%) and only slightly higher than previous estimates for Central American populations based on RAPD variation (20%). Significant fine-scale spatial structure was found in all of the surveyed mahogany populations and is probably strongly influenced by the limited seed dispersal range of the species. Furthermore, a survey of progeny arrays from selected mother trees in two of the plots indicated that most pollinations involved proximate trees. These data indicate that very little gene flow, via either pollen or seed, is occurring between blocks of mahogany within a continuous or disturbed forest landscape. Thus, once diversity is removed from a forest population of mahogany, these data suggest that recovery would be difficult via seed or pollen dispersal, and provides an explanation for mahogany's apparent susceptibility to the pressures of logging. Evidence is reviewed from other studies of gene flow and seedling regeneration to discuss alternative extraction strategies that may maintain diversity or allow recovery of genetic resources. PMID- 12634812 TI - Genomes of mice and men. PMID- 12634813 TI - Rose-colored goggles. PMID- 12634817 TI - One gene determines maize B chromosome accumulation by preferential fertilisation; another gene(s) determines their meiotic loss. AB - Genotypes of high (H(m)) and low (L(m)) male B transmission rate (B-TR) were obtained. B-TR segregation in the F2 is reported, showing that the H(m) and L(m) lines differ in a single locus we call mBt (male B transmission), controlling B preferential fertilisation in maize. The egg cells control which one of the sperm nuclei is going to fertilise them, mBt(h) egg cells being preferentially fertilised by the sperm nucleus carrying the supernumerary B chromosomes (Bs). It is hypothesised that the mBt gene is involved in the normal fertilisation of maize but the parasitic Bs take advantage of the mBt(h) allele to increase their own transmission. Selection was also carried out when the Bs were transmitted on the female side (H(f) and L(f) lines). The F1 hybrids show that the gene(s) that we call fBt (female B transmission), controlling female B-TR, is located on the A chromosomes acting at diploid level, the fBt(l) allele(s) for low transmission being dominant. This allele causes the loss of Bs at meiosis, which is shown using a specific B molecular probe to determine B presence/absence in microspores of both lines and hybrids. Maize Bs are a nice example of intragenome conflict, because the mBt and fBt loci are a polymorphic system of attack and defence between A and B chromosomes. PMID- 12634818 TI - Molecular genetic evidence for parthenogenesis in the Burmese python, Python molurus bivittatus. AB - Parthenogenesis among reptiles is rare. Only a few species have the ability to reproduce asexually. Most of these are obligate parthenogenetic species that consist (almost) entirely of females, which can reproduce solely through parthenogenesis. Rarer are sexual species that only sporadically reproduce through parthenogenesis. A female Python molurus bivittatus (Reptilia, Boidae) from the Artis Zoo, Amsterdam, produced eggs in five consecutive years that contained embryos while she was isolated from males. These eggs might be fertilized with stored sperm, or might be the product of parthenogenesis. Parthenogenesis has not been shown for the Boidae family before. We performed parentship analyses on the snake and seven of her embryos using microsatellites and AFLP. Four microsatellite loci developed for this species combined with three loci developed previously for different snake species revealed too little variation to discriminate between sperm retention and parthenogenesis. With AFLP we were able to confirm that the Artis Zoo female reproduced parthenogenetically. Because the offspring are genetically identical to their mother, whereas in previous studies on sporadic parthenogenesis in snakes a loss of genetic information was reported, we conclude that the meiotic pathways that produce the diploid egg cells are different. PMID- 12634819 TI - Synaptonemal complex karyotype of zebrafish. AB - Meiotic cells of zebrafish have been prepared to show synaptonemal complexes (SCs) by light and electron microscopy. Completely paired SCs from both spermatocytes and oocytes were measured to produce an SC karyotype. The SC karyotype resembles the somatic karyotype of zebrafish and has no recognisable sex bivalent. Measurements of total SC length indicate that SCs grow longer and develop centromeres during pachytene. Oocytes consistently have longer SCs than spermatocytes, presumably correlated with the reported higher recombination frequency in females than in males. PMID- 12634820 TI - Host-plant-associated genetic differentiation in Northern French populations of the European corn borer. AB - The phytophagous insects that damage crops are often polyphagous, feeding on several types of crop and on weeds. The refuges constituted by noncrop host plants may be useful in managing the evolution in pest species of resistance to the Bacillus thuringiensis toxins produced by transgenic crops. However, the benefits of these refuges may be limited because host-plant diversity may drive genetic divergence and possibly even host-plant-mediated sympatric speciation. The European corn borer, Ostrinia nubilalis Hubner (Lepidoptera: Crambidae), is the main pest of maize in Europe and North America, where it was introduced early in the 20th century. It has a wide host range but feeds principally on mugwort (Artemisia vulgaris L.) and maize (Zea mays L.). O. nubilalis is found on mugwort only in the northern part of France, whereas it is found on maize throughout France. The extent of genetic variation at allozyme markers was investigated in populations collected from the two host plants over the entire geographical distribution of the European corn borer on mugwort in France. Allelic differentiation between pairs of populations and hierarchical analyses of pools of samples from each host plant indicate that the group of populations feeding on maize differed from the group of populations feeding on mugwort. Our results suggest (1) host-plant-related divergent selection at the genomic region surrounding the Mpi locus and (2) limited gene flow between the populations feeding on mugwort and those infesting maize fields. These data indicate that adults emerging from mugwort would not be useful for managing the evolution of resistance to the B. thuringiensis toxins in European corn borer populations. PMID- 12634821 TI - Genetic effects on microsatellite diversity in wild emmer wheat (Triticum dicoccoides) at the Yehudiyya microsite, Israel. AB - This study investigated allele size constraints and clustering, and genetic effects on microsatellite (simple sequence repeat, SSR) diversity at 28 loci comprising seven types of tandem repeated dinucleotide motifs in a natural population of wild emmer wheat, Triticum dicoccoides, from a shade vs sun microsite in Yehudiyya, northeast of the Sea of Galilee, Israel. It was found that allele distribution at SSR loci is clustered and constrained with lower or higher boundary. This may imply that SSR have functional significance and natural constraints. Genetic factors, involving genome, chromosome, motif, and locus significantly affected SSR diversity. Genome B appeared to have a larger average repeat number (ARN), but lower variance in repeat number (sigma(ARN)(2)), and smaller number of alleles per locus than genome A. SSRs with compound motifs showed larger ARN than those with perfect motifs. The effects of replication slippage and recombinational effects (eg, unequal crossing over) on SSR diversity varied with SSR motifs. Ecological stresses (sun vs shade) may affect mutational mechanisms, influencing the level of SSR diversity by both processes. PMID- 12634822 TI - Wolbachia segregation dynamics and levels of cytoplasmic incompatibility in Drosophila sechellia. AB - In Drosophila sechellia, the endocellular bacterium Wolbachia induces cytoplasmic incompatibility (CI): in crosses involving infected males, a partial or complete embryonic mortality occurs unless the female bears the same Wolbachia. D. sechellia is known to harbour two Wolbachia variants, namely wSh and wSn, closely related to wHa and wNo, respectively, two strains infecting the populations of D. simulans from the Seychelles archipelago and New Caledonia. Strikingly, the two species show similar infection patterns: in D. sechellia, wSh can be present on its own or in double infection with wSn, but individuals carrying wSn only do not occur; in D. simulans, wHa can be present on its own or in double infection with wNo, but individuals carrying wNo only do not occur, or occur at very low frequency. Previous experiments on D. simulans showed that lines singly infected by wNo can be obtained by segregation, and stably maintained. Here we investigate this issue in D. sechellia through an 18 generation experiment, and show that wSn and wSh singly infected lines can arise by segregation. Using singly infected lines obtained in this experiment, we estimate the CI intensities of wSh and wSn in D. sechellia, and compare these to the CI intensities of the same Wolbachia injected into D. simulans. Our results do not suggest any consistent effect of the host species on the CI induced by wSh. On the contrary, it seems that wSn expression is repressed by host factors in D. sechellia. PMID- 12634823 TI - On migration load of seeds and pollen grains in a local population. AB - We have extended Wright's model of migration load to hermaphrodite plants showing variation at a single locus with two alleles. The model incorporates independent migration of seeds and pollen grains, the selection at both the haploid gametophyte and the diploid sporophyte stages, and a mixed mating system. The analytical relations between migration load and migration rate of seeds and pollen grains are explicitly formulated. The results show that under certain conditions, seed flow can have a more effect on migration load than pollen flow. Pollen selection at the gametophyte stage cannot substantially affect the migration load at the sporophyte stage. Selection at the diploid sporophyte stage is critical in determining the migration load of pollen grains. The relative migration loads of pollen versus seeds can be approximately estimated in predominantly outcrossing populations by the ratio of pollen flow to twice the seed flow, when the selection coefficient (s(T)) is greater than, or approximately equal to, the migration rate (m). PMID- 12634824 TI - Linkage mapping in tetraploid willows: segregation of molecular markers and estimation of linkage phases support an allotetraploid structure for Salix alba x Salix fragilis interspecific hybrids. AB - Salix alba-Salix fragilis complex includes closely related dioecious polyploid species, which are obligate outcrossers. Natural populations of these willows and their hybrids are represented by a mixture of highly heterozygous genotypes sharing a common gene pool. Since nothing is known about their genomic constitution, tetraploidy (2n=4x=76) in willow species makes basic and applied genetic studies difficult. We have used a two-way pseudotestcross strategy and single-dose markers (SDMs) to construct the first linkage maps for both pistillate and staminate willows. A total of 242 amplified fragment length polymorphisms (AFLPs) and 50 selective amplifications of microsatellite polymorphic loci (SAMPL) markers, which showed 1:1 segregation in the F(1) mapping populations, were used in linkage analysis. In S. alba, 73 maternal and 48 paternal SDMs were mapped to 19 and 16 linkage groups covering 708 and 339 cM, respectively. In S. fragilis, 13 maternal and 33 paternal SDMs were mapped in six and 14 linkage groups covering 98 and 321 cM, respectively. For most cosegregation groups, a comparable number of markers linked in coupling and repulsion was identified. This finding suggests that most of chromosomes pair preferentially as occurs in allotetraploid species exhibiting disomic inheritance. The detection of 10 pairs of marker alleles from single parents showing codominant inheritance strengthens this hypothesis. The fact that, of the 1122 marker loci identified in the two male and female parents, the vast majority (77.5%) were polymorphic and as few as 22.5% were shared between parental species highlight that S. alba and S. fragilis genotypes are differentiated. The highly difference between S. alba- and S. fragilis-specific markers found in both parental combinations (on average, 65.3 vs 34.7%, respectively) supports the (phylogenetic) hypothesis that S. fragilis is derived from S. alba-like progenitors. PMID- 12634825 TI - Genetic and demographic dynamics of small populations of Silene latifolia. AB - Small local populations of Silene alba, a short-lived herbaceous plant, were sampled in 1994 and again in 1999. Sampling included estimates of population size and genetic diversity, as measured at six polymorphic allozyme loci. When averaged across populations, there was very little change between samples (about three generations) in population size, measures of within-population genetic diversity such as number of alleles or expected heterozygosity, or in the apportionment of genetic diversity within and among populations as measured by F(st). However, individual populations changed considerably, both in terms of numbers of individuals and genetic composition. Some populations doubled in size between samples, while others shrank by more than 75%. Similarly, expected heterozygosity and allele number increased by more than two-fold in individual populations and decreased by more than three-fold in others. When population specific change in number and change in measures of genetic diversity were considered together, significant positive correlations were found between the demographic and genetic variables. It is speculated that some populations were released from the demographic consequences of inbreeding depression by gene flow. PMID- 12634826 TI - A multiyear estimate of the effective pollen donor pool for Albizia julibrissin. AB - Studies of pollen movement in plant populations are often limited to a single reproductive event, despite concerns about the adequacy of single-year measures for perennial organisms. In this study, we estimate the effective number of pollen donors per tree from a multiyear study of Albizia julibrissin Durazz (mimosa, Fabaceae), an outcrossing, insect-pollinated tree. We determined 40 seedling genotypes for each of 15 seed trees during 4 successive years. A molecular analysis of variance of the pollen gametes fertilizing the sampled seeds was used to partition variation in pollen pools among seed trees, among years, and within single tree-year collections. Using these variance components, we demonstrate significant male gametic variability among years for individual trees. However, results indicate that yearly variation in the 'global pollen pool', averaged over all 15 seed trees for these 4 years, is effectively zero. We estimate the effective number of pollen donors for a single mimosa tree (N(ep)) to be 2.87. Single season analyses yield N(ep) approximately 2.05, which is 40% less than the value of N(ep) estimated from 4 years of data. We discuss optimal sampling for future studies designed to estimate N(ep). Studies should include more trees, each sampled over at least a few years, with fewer seeds per tree per year than are needed for a traditional parentage study. PMID- 12634827 TI - Thermal tolerance trade-offs associated with the right arm of chromosome 3 and marked by the hsr-omega gene in Drosophila melanogaster. AB - Drosophila melanogaster occurs in diverse climatic regions and shows opposing clinal changes in resistance to heat and resistance to cold along a 3000 km latitudinal transect on the eastern coast of Australia. We report here on variation at a polymorphic 8 bp-indel site in the heat shock hsr-omega gene that maps to the right arm of chromosome 3. The frequency of the genetic element marked by the L form of the gene was strongly and positively associated with latitude along this transect, and latitudinal differences in L frequency were robustly associated with latitudinal differences in maximum temperature for the hottest month. On a genetic background mixed for genes from each end of the cline a set of 10 lines was derived, five of which were fixed for the L marker, the absence of In(3R)P and 12 kb of repeats at a second polymorphic site at the 3' end of hsr-omega, and five that were fixed for the S marker, In(3R)P and 15 kb of hsr-omega repeats. For two different measures of heat tolerance S lines outperformed L lines, and for two different measures of cold tolerance L lines outperformed S lines. These data suggest that an element on the right arm of chromosome 3, possibly In(3R)P, confers heat resistance but carries the trade-off of also conferring susceptibility to cold. This element occurs at high frequency near the equator. The alternate element on the other hand, at high frequency at temperate latitudes, confers cold resistance at the cost of heat susceptibility. PMID- 12634830 TI - A Faustian bargain for universities? PMID- 12634831 TI - Science and culture. Western science could learn a thing or two from the way science is done in other cultures. PMID- 12634832 TI - Worms and science. An interview with Sydney Brenner, distinguished research professor at The Salk Institute, La Jolla, USA, and one of the winners of 2002 Nobel Prize for Physiology and Medicine. PMID- 12634833 TI - Biotechnology to fight bioterrorism. While the fight against terrorism is helping to revitalize the biotech market, academic scientists are starting to worry about limitations on research. PMID- 12634834 TI - Feeding prejudice. Reluctance within the European Union to accept genetically modified crops may hinder the benefits of this technology reaching the developing world. PMID- 12634835 TI - DNA and consumer confidence. DNA fingerprinting and DNA-based labelling systems are gaining importance in the security market to verify the authenticity of products. PMID- 12634838 TI - Mont Sainte-Odile: a sanctuary for GPCRs. Confidence on signal transduction of G protein-couple receptors. PMID- 12634839 TI - Coupling adhesion to actin bundles in the inner ear. Novel functions for novel cadherins. PMID- 12634840 TI - The Ark1/Prk1 family of protein kinases. Regulators of endocytosis and the actin skeleton. AB - The Ark/Prk serine/threonine kinases initiate phosphorylation cycles that control the endocytic machinery in mammalian cells and in yeast, and the actin cytoskeleton in yeast. The members of this protein family are unified by homologies in their kinase domain, but are generally diverse in their other domains. The evolution of Ark/Prk family members in different organisms may have allowed the conserved role of the kinase domain, which is required for the phosphorylation of both endocytic and cytoskeletal components, to be coupled to other functional domains. PMID- 12634841 TI - Multiple roles for kinases in DNA replication. AB - DNA replication is carried out by the replisome, which includes several proteins that are targets of cell-cycle-regulated kinases. The phosphorylation of proteins such as replication protein A, DNA polymerase-alpha and -delta, replication factor C, flap endonuclease 1 and DNA ligase I leads to their inactivation, suggesting that phosphorylation is important in the prevention of re-replication. Moreover, the phosphorylation of several of these replication proteins has been shown to block their association with the 'moving platform'-proliferating cell nuclear antigen. Therefore, phosphorylation seems to be a crucial regulator of replisome assembly and DNA replication, although its precise role in these processes remains to be clarified. PMID- 12634842 TI - Segments missing from the draft human genome sequence can be isolated by transformation-associated recombination cloning in yeast. AB - The reported draft human genome sequence includes many contigs that are separated by gaps of unknown sequence. These gaps may be due to chromosomal regions that are not present in the Escherichia coli libraries used for DNA sequencing because they cannot be cloned efficiently, if at all, in bacteria. Using a yeast artificial chromosome (YAC)/ bacterial artificial chromosome (BAC) library generated in yeast, we found that approximately 6% of human DNA sequences tested transformed E. coli cells less efficiently than yeast cells, and were less stable in E. coli than in yeast. When the ends of several YAC/BAC isolates cloned in yeast were sequenced and compared with the reported draft sequence, major inconsistencies were found with the sequences of those YAC/BAC isolates that transformed E. coli cells inefficiently. Two human genomic fragments were re isolated from human DNA by transformation-associated recombination (TAR) cloning. Re-sequencing of these regions showed that the errors in the draft are the results of both missassembly and loss of specific DNA sequences during cloning in E. coli. These results show that TAR cloning might be a valuable method that could be widely used during the final stages of the Human Genome Project. PMID- 12634843 TI - Bidirectional DNA unwinding by a ternary complex of T antigen, nucleolin and topoisomerase I. AB - The simian virus 40 large tumour-antigen (T antigen) DNA helicase is a hexameric structure; it has been proposed that, in viral DNA replication, two of these hexamers are combined to form a bipartite holoenzyme that acts concurrently at both forks of a replication bubble. In a search for structural components of this helicase complex, we have identified nucleolin as a specific binding protein for the T-antigen hexamer. We show that nucleolin, in co-operation with human topoisomerase I, mediates the cohesion of the T-antigen helicase holoenzyme during plasmid unwinding. Our results provide biochemical evidence for a direct role of nucleolin in DNA replication, in addition to its known function in ribosome biogenesis. The data presented here suggest that nucleolin enables the formation of a functional 'helicase-swivelase' complex at the replication fork. PMID- 12634844 TI - Erroneous incorporation of oxidized DNA precursors by Y-family DNA polymerases. AB - Deranged oxidative metabolism is a property of many tumour cells. Oxidation of the deoxynucleotide triphosphate (dNTP) pool, as well as DNA, is a major cause of genome instability. Here, we report that two Y-family DNA polymerases of the archaeon Sulfolobus solfataricus strains P1 and P2 incorporate oxidized dNTPs into nascent DNA in an erroneous manner: the polymerases exclusively incorporate 8-OH-dGTP opposite adenine in the template, and incorporate 2-OH-dATP opposite guanine more efficiently than opposite thymine. The rate of extension of the nascent DNA chain following on from these incorporated analogues is only slightly reduced. These DNA polymerases have been shown to bypass a variety of DNA lesions. Thus, our results suggest that the Y-family DNA polymerases promote mutagenesis through the erroneous incorporation of oxidized dNTPs during DNA synthesis, in addition to facilitating translesion DNA synthesis. We also report that human DNA polymerase eta, a human Y-family DNA polymerase, incorporates the oxidized dNTPs in a similar erroneous manner. PMID- 12634845 TI - The soybean retroelement SIRE1 uses stop codon suppression to express its envelope-like protein. AB - The soybean SIRE1 family of Ty1/copia retrotransposons encodes an envelope-like gene (env-like). We analysed the DNA sequences of nine SIRE1 insertions and observed that the gag/pol and env-like genes are in the same reading frame and separated by a single UAG stop codon. The six nucleotides immediately downstream of the stop codon conform to a degenerate nucleotide motif, CARYYA, which is sufficient to facilitate stop codon suppression in tobacco mosaic virus. In vivo stop codon suppression assays indicate that SIRE1 sequences confer leakiness to the UAG stop codon at an efficiency of 5%. These data suggest that SIRE1 retro elements use translational suppression to express their envelope-like protein; this is in contrast with all characterized retroviruses, which express the envelope protein from a spliced genomic messenger RNA. PMID- 12634846 TI - Identification of Gbp2 as a novel poly(A)+ RNA-binding protein involved in the cytoplasmic delivery of messenger RNAs in yeast. AB - Important progress in understanding messenger RNA export from the nucleus could be achieved by increasing the list of proteins that are involved in this process. Here, we present the identification of Gbp2 as a novel shuttling RNA-binding protein in Saccharomyces cerevisiae. Nuclear import of Gbp2 is dependent on the receptor Mtr10 and the serine/arginine-specific protein kinase Sky1. Deletion of the genes encoding both of these proteins or disruption of two of the arginine/serine repeats each shifts the steady-state localization of Gbp2 to the cytoplasm. Interestingly, deletion of MTR10 only also causes an increase in poly(A)(+) RNA binding by Gbp2, suggesting a role of Mtr10 in the dissociation of Gbp2 from mRNA in the cytoplasm. The nuclear export of Gbp2 is always coupled to mRNA export and is dependent on continuous RNA polymerase II transcription and mRNA-export factors. Although GBP2 is not essential for normal cell growth, overexpression of this gene is toxic and causes a nuclear retention of bulk poly(A)(+) RNA. Together, our findings clearly show an involvement of Gbp2 in mRNA transport. In addition, as a non-essential protein, Gbp2 also has the interesting potential to be spatially or temporally regulated. PMID- 12634847 TI - RNA chaperone activity of the Sm-like Hfq protein. AB - The Escherichia coli Sm-like host factor I (Hfq) protein is thought to function in post-transcriptional regulation by modulating the function of small regulatory RNAs. Hfq also interferes with ribosome binding on E. coli ompA messenger RNA, indicating that Hfq also interacts with mRNAs. In this study, we have used stimulation of group I intron splicing in vivo and a modified in vitro toeprinting assay to determine whether Hfq acts as an RNA chaperone. Hfq was able to rescue an RNA 'folding trap' in a splicing defective T4 bacteriophage td gene in vivo. Enzymatic analysis showed that Hfq affects the accessibility of the ompA start codon, as well as other bases within the ribosome-binding site, explaining its negative effect on ribosome binding. We also show that the Hfq-induced structural changes in ompA mRNA are maintained after proteolytic digestion of the protein, which classifies Hfq as an RNA chaperone. PMID- 12634848 TI - The 37 kDa/67 kDa laminin receptor is required for PrP(Sc) propagation in scrapie infected neuronal cells. AB - The accumulation of PrP(Sc) in scrapie-infected neuronal cells has been prevented by three approaches: (i) transfection of ScMNB cells with an antisense laminin receptor precursor (LRP) RNA-expression plasmid, (ii) transfection of ScN2a cells and ScGT1 cells with small interfering RNAs (siRNAs) specific for the LRP mRNA, and (iii) incubation of ScN2a cells with an anti-LRP/LR antibody. LRP antisense RNA and LRP siRNAs reduced LRP/LR expression and inhibited the accumulation of PrP(Sc) in these cells. The treatments also reduced PrP(c) levels. The anti LRP/LR antibody, W3, abolished PrP(Sc) accumulation and reduced PrP(c) levels after seven days of incubation. Cells remained free of PrP(Sc) after being cultured for 14 additional days without the antibody, whereas the PrP(c) level was restored. Our results demonstrate the necessity of the laminin receptor (LRP/LR) for PrP(Sc) propagation in cultured cells and suggest that LRP/LR specific antibodies could be used as powerful therapeutic tools in the treatment of transmissible spongiform encephalopathies. PMID- 12634849 TI - Occurrence of a putative ancient-like isomerase involved in histidine and tryptophan biosynthesis. AB - We report the occurrence of an isomerase with a putative (betaalpha)(8)-barrel structure involved in both histidine and trypto-phan biosynthesis in Streptomyces coelicolor A3(2) and Mycobacterium tuberculosis HR37Rv. Deletion of a hisA homologue (SCO2050) putatively encoding N'-[(5'-phosphoribosyl)-formimino]-5 amino-imidazole-4-carboxamide ribonucleotide isomerase from the chromosome of S. coelicolor A3(2) generated a double auxotrophic mutant for histidine and tryptophan. The bifunctional gene SCO2050 and its orthologue Rv1603 from M. tuberculosis complemented both hisA and trpF mutants of Escherichia coli. Expression of the E. coli trpF gene in the S. coelicolor mutant only complemented the tryptophan auxo-trophy, and the hisA gene only complemented the histidine auxotrophy. The discovery of this enzyme, which has a broad-substrate specificity, has implications for the evolution of metabolic pathways and may prove to be important for understanding the evolution of the (betaalpha)(8) barrels. PMID- 12634850 TI - Parkin binds the Rpn10 subunit of 26S proteasomes through its ubiquitin-like domain. AB - Parkin, a product of the causative gene of autosomal-recessive juvenile parkinsonism (AR-JP), is a RING-type E3 ubiquitin ligase and has an amino terminal ubiquitin-like (Ubl) domain. Although a single mutation that causes an Arg to Pro substitution at position 42 of the Ubl domain (the Arg 42 mutation) has been identified in AR-JP patients, the function of this domain is not clear. In this study, we determined the three-dimensional structure of the Ubl domain of parkin by NMR, in particular by extensive use of backbone (15)N-(1)H residual dipolar-coupling data. Inspection of chemical-shift-perturbation data showed that the parkin Ubl domain binds the Rpn10 subunit of 26S proteasomes via the region of parkin that includes position 42. Our findings suggest that the Arg 42 mutation induces a conformational change in the Rpn10-binding site of Ubl, resulting in impaired proteasomal binding of parkin, which could be the cause of AR-JP. PMID- 12634851 TI - Genetic inactivation of Par4 results in hyperactivation of NF-kappaB and impairment of JNK and p38. AB - The Par4 gene was first identified in prostate cells undergoing apoptosis after androgen withdrawal. PAR4 was subsequently shown to interact with, and inhibit, atypical protein kinase C isoforms, functioning as a negative regulator of the NF kappaB pathway. This may explain its pro-apoptotic function in overexpression experiments. To determine the physiological role of PAR4, we have derived primary embryonic fibroblasts (EFs) from Par4(-/-) mice. We show here that loss of PAR4 leads to a reduction in the ability of tumour necrosis factor-alpha (TNF-alpha) to induce apoptosis by increased activation of NF-kappaB. Consistent with recent reports demonstrating the antagonistic actions of NF-kappaB and c-Jun amino terminal kinase (JNK) signalling, we have found that Par4(-/-) cells show a reduced activation of the sustained phase of JNK and p38 stimulation by TNF-alpha and interleukin 1. Higher levels of an anti-apoptotic JNK-inhibitor protein, X chromosome-linked inhibitor of apoptosis, in Par4(-/-) EFs might explain the inhibition of JNK activation in these cells. PMID- 12634852 TI - Dynamics of the interaction between the insulin receptor and protein tyrosine phosphatase 1B in living cells. AB - The dynamics of the interaction of the insulin receptor with a substrate-trapping mutant of protein-tyrosine phosphatase 1B (PTP1B) were monitored in living human embryonic kidney cells using bioluminescence resonance energy transfer (BRET). Insulin dose-dependently stimulates this interaction, which could be followed in real time for more than 30 minutes. The effect of insulin on the BRET signal could be detected at early time-points (30 seconds), suggesting that in intact cells the tyrosine-kinase activity of the insulin receptor is tightly controlled by PTP1B. Interestingly, the basal (insulin-independent) interaction of the insulin receptor with PTP1B was much weaker with a soluble form of the tyrosine phosphatase than with the endoplasmic reticulum (ER)-targeted form. Inhibition of insulin-receptor processing using tunicamycin suggests that the basal interaction occurs during insulin-receptor biosynthesis in the ER. Therefore, localization of PTP1B in this compartment might be important for the regulation of insulin receptors during their biosynthesis. PMID- 12634853 TI - The membrane-tethering protein p115 interacts with GBF1, an ARF guanine nucleotide-exchange factor. AB - The membrane-transport factor p115 interacts with diverse components of the membrane-transport machinery. It binds two Golgi matrix proteins, a Rab GTPase, and various members of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) family. Here, we describe a novel interaction between p115 and Golgi-specific brefeldin-A-resistant factor 1 (GBF1), a guanine nucleotide exchange factor for ADP ribosylation factor (ARF). GBF1 was identified in a yeast two-hybrid screen, using full-length p115 as bait. The interaction was confirmed biochemically, using in vitro and in vivo assays. The interacting domains were mapped to the proline-rich region of GBF1 and the head region of p115. These proteins colocalize extensively in the Golgi and in peripheral vesicular tubular clusters. Mutagenesis analysis indicates that the interaction is not required for targeting GBF1 or p115 to membranes. Expression of the p115 binding (pro-rich) region of GBF1 leads to Golgi disruption, indicating that the interaction between p115 and GBF1 is functionally relevant. PMID- 12634855 TI - On the role of CFTR, PSSR1 and PST1/SPINK1 in idiopathic chronic pancreatitis. PMID- 12634857 TI - No evidence for a parental inversion polymorphism predisposing to rearrangements at 22q11.2 in the DiGeorge/Velocardiofacial syndrome. PMID- 12634858 TI - Genome-wide search in Finnish families with inflammatory bowel disease provides evidence for novel susceptibility loci. AB - Epidemiological and genetic linkage studies have indicated a strong genetic basis for development of inflammatory bowel disease (IBD) which was recently supported by discovery of the Crohn's disease (CD) susceptibility gene termed NOD2/CARD15. We carried out a genome-wide linkage study in Finnish IBD families, providing a particular advantage to map susceptibility genes for ulcerative colitis (UC) within a genetic isolate. Initially, 92 IBD families with 138 affected sib-pairs (ASPs), were genotyped for 429 markers spaced at approximately 10 cM intervals. Next, the loci on chromosomes 2p13-11, 11p12-q13, and 12p13-12 were high-density mapped in the extended family cohort of 130 families with 173 ASPs. In this study, the most significant lod scores were observed for the UC families on chromosome 2p11 (D2S2333), in the vicinity of the REG gene cluster which is strikingly overexpressed in the IBD mucosa. The maximum two-point lod score was 3.34 (dominant model), and the corresponding NPL score 2.61. For UC, the second highest two-point NPL score of 2.00 was observed at proximal 12p13, where also some evidence for linkage disequilibrium emerged (P=0.07 and P=0.007 for the basic and extended IBD cohorts, respectively). The highest two-point NPL score for the CD families was 2.34 at D12S78 (12q23) with significant evidence for linkage disequilibrium (P=0.004), and for the mixed (MX) families 2.07 at D4S406 near the linkage peak reported previously. This study confirmed several of the IBD loci that have previously been reported and gives evidence for new IBD loci on chromosomes 2p11, 11p12-q13, 12p13-12, 12q23, and 19q13. PMID- 12634859 TI - A new locus (DFNA47) for autosomal dominant non-syndromic inherited hearing loss maps to 9p21-22 in a large Italian family. AB - Hearing loss is the most common sensory disorder in humans, and genetic factors are a major cause. Approximately 15-20% of genetic cases exhibit an autosomal dominant pattern of transmission. So far, 41 autosomal dominant loci have been mapped and 17 genes have been identified. Here we report the mapping of a novel locus for autosomal dominant non-syndromic hearing loss, DFNA47, to chromosome 9p21-22 in a large multigenerational Italian family with progressive hearing impairment. Most affected individuals noticed hearing impairment after their teens with subsequent gradual progression to a moderate-severe loss. There were no obvious vestibular dysfunction and other associated abnormalities. A maximum lod score of 3.14 was obtained with marker D9S157 (at theta=0) after a genome wide search. The study of additional markers allowed us to confirm this region with positive lod scores of 3.58 (at theta=0 from D9S285) and of 3.67 (at theta=0 from D9S162). Recombinants define a region of approximately 9 cM flanked by markers D9S268 and D9S942. Multipoint linkage analysis showed a Lod score of 4.26. Few known genes map to the region, and those possibly related by function to hearing are being screened for disease-causing mutations. PMID- 12634860 TI - Combined high resolution linkage and association mapping of quantitative trait loci. AB - In this paper, we investigate variance component models of both linkage analysis and high resolution linkage disequilibrium (LD) mapping for quantitative trait loci (QTL). The models are based on both family pedigree and population data. We consider likelihoods which utilize flanking marker information, and carry out an analysis of model building and parameter estimations. The likelihoods jointly include recombination fractions, LD coefficients, the average allele substitution effect and allele dominant effect as parameters. Hence, the model simultaneously takes care of the linkage, LD or association and the effects of the putative trait locus. The models clearly demonstrate that linkage analysis and LD mapping are complementary, not exclusive, methods for QTL mapping. By power calculations and comparisons, we show the advantages of the proposed method: (1) population data can provide information for LD mapping, and family pedigree data can provide information for both linkage analysis and LD mapping; (2) using family pedigree data and a sparse marker map, one may investigate the prior suggestive linkage between trait locus and markers to obtain low resolution of the trait loci, because linkage analysis can locate a broad candidate region; (3) with the prior knowledge of suggestive linkage from linkage analysis, both population and family pedigree data can be used simultaneously in high resolution LD mapping based on a dense marker map, since LD mapping can increase the resolution for candidate regions; (4) models of high resolution LD mappings using two flanking markers have higher power than that of models of using only one marker in the analysis; (5) excluding the dominant variance from the analysis when it does exist would lose power; (6) by performing linkage interval mappings, one may get higher power than by using only one marker in the analysis. PMID- 12634861 TI - The epsilon-sarcoglycan gene (SGCE), mutated in myoclonus-dystonia syndrome, is maternally imprinted. AB - Myoclonus-dystonia syndrome (MDS) is a non-degenerative neurological disorder that has been described to be inherited in an autosomal dominant mode with incomplete penetrance. MDS is caused by loss of function mutations in the epsilon sarcoglycan gene. Reinvestigation of MDS pedigrees provided evidence for a maternal imprinting mechanism. As differential methylated regions (DMRs) are a characteristic feature of imprinted genes, we studied the methylation pattern of CpG dinucleotides within the CpG island containing the promoter region and the first exon of the SGCE gene by bisulphite genomic sequencing. Our findings revealed that in peripheral blood leukocytes the maternal allele is methylated, while the paternal allele is unmethylated. We also showed that most likely the maternal allele is completely methylated in brain tissue. Furthermore, CpG dinucleotides in maternal and paternal uniparental disomy 7 (UPD7) lymphoblastoid cell lines show a corresponding parent-of-origin specific methylation pattern. The effect of differential methylation on the expression of the SGCE gene was tested in UPD7 cell lines with only a weak RT-PCR signal observed in matUPD7 and a strong signal in patUPD7. These results provide strong evidence for a maternal imprinting of the SGCE gene. The inheritance pattern in MDS families is in agreement with such an imprinting mechanism with the exception of a few cases. We investigated one affected female that inherited the mutated allele from her mother. Surprisingly, we found the paternal wild type allele expressed whereas the mutated maternal allele was not detectable in peripheral blood cDNA. PMID- 12634862 TI - Familial juvenile hyperuricaemic nephropathy (FJHN): linkage analysis in 15 families, physical and transcriptional characterisation of the FJHN critical region on chromosome 16p11.2 and the analysis of seven candidate genes. AB - Familial juvenile hyperuricaemic nephropathy (FJHN) is an autosomal dominant renal disease characterised by juvenile onset of hyperuricaemia, gouty arthritis, and progressive renal failure at an early age. Recent studies in four kindreds showed linkage of a gene for FJHN to the same genomic interval on chromosome 16p11.2, where the gene for the phenotypically similar medullary cystic disease type 2 (MCKD2) has been localised. In this study we performed linkage analysis in additional 15 FJHN families. Linkage of FJHN to 16p11.2 was confirmed in six families, which suggests that, in a large proportion of FJHN kindreds, the disease is likely to be caused by a gene or genes located outside of 16p11.2. Haplotype analysis of the new and previously analysed families provided two non overlapping critical regions on 16p11.2-FJHN1, delimited by markers D16S499 D16S3036 and FJHN2, delimited by markers D16S412-D16S3116. Considering MCKD2 to be a distinct molecular entity, the analysis suggests that as many as three kidney disease genes may be located in close proximity on 16p11.2. From genomic databases we compiled integrated physical and transcription maps of whole critical genomic region in which 45 known genes and 129 predicted loci have been localised. We selected, analysed and found no pathogenic mutations in seven candidate genes. The linkage and haplotype analysis reported here demonstrates the genetic heterogeneity of FJHN. The report of integrated physical and mostly in-silico predicted transcription maps of the FJHN critical region provides a basis for precise experimental annotation of the current transcript map, which is essential for final identification of the FJHN gene(s). PMID- 12634863 TI - Identification and characterisation of the retinitis pigmentosa 1-like1 gene (RP1L1): a novel candidate for retinal degenerations. AB - Retinitis pigmentosa (RP) is the most common form of inherited retinopathy, with an approximate incidence of 1 in 3700 individuals worldwide. Mutations in the retinitis pigmentosa 1 (RP1) gene are responsible for about 5-10% cases of autosomal dominant RP. The RP1 gene is specifically expressed in the photoreceptor layers of the postnatal retina and encodes a predicted protein characterised by the presence of two doublecortin (DC) domains, known to be implicated in microtubule binding. We identified and characterised, both in human and in mouse, a novel mammalian gene, termed Retinitis Pigmentosa1-like1 (RP1L1), because of its significant sequence similarity to the RP1 gene product. The sequence homology between RP1 and RP1L1 was found to be mostly restricted to the DC domains and to the N-terminal region, including the first 350 amino acids. The RP1L1 gene was also found to be conserved in distant vertebrates, since we identified a homologue in Fugu rubripes (pufferfish). Similar to RP1, RP1L1 expression is restricted to the postnatal retina, as determined by semiquantitative reverse transcriptase-PCR and Northern analysis. The retina specific expression and the sequence similarity to RP1 render RP1L1 a potential candidate for inherited retinal disorders. PMID- 12634864 TI - Screening for PAX6 gene mutations is consistent with haploinsufficiency as the main mechanism leading to various ocular defects. AB - PAX6, a paired box transcriptional factor, is considered as the master control gene for morphogenesis of the eye. Human PAX6 mutations have been associated with a range of eye abnormalities, including aniridia, various anterior segment defects and foveal hypoplasia. We carried out a mutational analysis of the PAX6 gene in 54 unrelated patients with aniridia or related syndromes. A deleterious variation was evidenced in 17 sporadic cases (50%) and in 13 (72%) familial cases. Twenty-four different mutations, 17 of which are novel, were found. The spectrum of PAX6 mutations was highly homogeneous: 23 mutations (96%) leading to premature stop codons (eight nonsense and four splice site mutations, 11 insertions and deletions) and only one (4%) missense mutation. Twenty-two mutations were associated with aniridia phenotypes whereas two were associated with atypical phenotypes. These latter encompassed a missense mutation (R19P) in an individual with a microphthalmia-sclerocornea and a splice site mutation (IVS4+5G > C) in a family presenting with a congenital nystagmus. Both represented the most probably hypomorphic alleles. Aniridia cases were associated with nonsense or frameshifting mutations. A careful examination of the phenotypes did not make it possible to recognise significant differences whenever the predicted protein was deprived of one or another of its functional domains. This strongly suggested that most of the truncating mutations generated null alleles by nonsense mediated mRNA decay. Our observations support the concept of dosage effects of the PAX6 mutations as well as presenting evidence for variable expressivity. PMID- 12634865 TI - A new quantitative PCR multiplex assay for rapid analysis of chromosome 17p11.2 12 duplications and deletions leading to HMSN/HNPP. AB - A 1.4-Mb tandem duplication, including the gene for peripheral myelin protein 22 (PMP22) in chromosome 17p11.2-12 is responsible for 70% of the cases of the demyelinating type 1 of Charcot-Marie-Tooth disease or hereditary motor and sensory neuropathy I (CMT1A/HMSN I). A reciprocal deletion of this CMT1A region causes the hereditary neuropathy with liability to pressure palsies (HNPP). The CMT1A duplication increases the PMP22 gene dosage from two to three, the HNPP deletion reduces the gene dosage from two to one. Currently, routine diagnosis of HMSN/HNPP patients is mainly performed with polymorphic markers in-between the repetitive elements flanking the CMT1A region. These show quantitative and/or qualitative changes in case of a CMT1A duplication and a homozygous allele pattern in case of HNPP deletion. In HNPP patients the deletion is usually confirmed by fluorescence in situ hybridisation (FISH). We now developed a reliable, single tube real-time quantitative PCR assay for rapid determination of PMP22 gene dosage directly. This method involves a multiplex reaction using FAM labelled Taqman-probe with TAMRA quencher derived from PMP22 exon 3 and a VIC labelled probe with non-fluorescent quencher from exon 12 of the albumin gene as internal reference. Copy number of the PMP22 gene was determined by the comparative threshold cycle method (deltadeltaCt). Each sample was run in quadruplicate and analysed at two different threshold levels. The level giving the smallest standard deviation was scored. We evaluated this method through the retrospective analysis of 252 HMSN patients with known genotype and could confirm the previous findings in 99% of cases. Two patients were wrongly diagnosed with microsatellite analysis while quantitative real-time PCR identified the correct genotype, as confirmed by FISH. Thus, this method shows superior sensitivity to microsatellite analysis and has the additional advantage of being a fast and uniform assay for quantitative analysis of both CMT1A and HNPP. PMID- 12634866 TI - Clustering of Crohn's disease within affected sibships. AB - Crohn's disease (CD) is a complex genetic disorder for which aetiology is unknown. Recently, genetic factors for susceptibility have been described. Several genetic loci have been mapped and partially explain the familial aggregations of the disease. However, environmental factors may also contribute to these aggregations. We considered that if the role of non-genetic factors was negligible, CD patients would be randomly distributed in sibships with multiple affected siblings. On the other hand if there was a significant environmental contribution, the siblings would be affected non-randomly over exposure status. In order to test this hypothesis, we studied 102 sibships with two or more affected siblings. A statistical test, named Cluster of Affected Sibling Test or CAST, was developed, based on the exact calculation of the probability of observing a given number of clusters of affected siblings in multiplex families. The null hypothesis of a random distribution of affected siblings was rejected (P=0.005). The observed excess of affected sibling clusters indicates that birth order influences the disease status. Considering that an adjacent order of birth is a global estimate of environmental sharing, this observation strongly suggests that environmental factors contribute to the observed familial aggregations of the disease. This observation provides evidence that familial CD is a relevant tool for further studies of environmental factors and gene-environment interaction. More generally, the CAST statistics may be widely applicable to estimate the involvement of environmental factors in the aetiology of other binary traits which may be observed in multiple members of the same sibship. PMID- 12634867 TI - Mapping of a new autosomal recessive nonsyndromic hearing loss locus (DFNB32) to chromosome 1p13.3-22.1. AB - Approximately 80% of the hereditary hearing loss is nonsyndromic. Isolated deafness is the most genetically heterogeneous trait. We have ascertained 10 individuals from a large consanguineous Tunisian family with congenital profound autosomal recessive deafness. All affected individuals are otherwise healthy. Genotype analysis excluded linkage to known recessive deafness loci in this family. Following a genome wide screening, a linkage was detected only with locus D1S206 on chromosome 1, thereby defining a novel deafness locus, DFNB32. In order to confirm linkage and for fine mapping the genetic interval, 12 individuals belonging to this family were added and 19 microsatellite markers were tested. A maximum two-point lodscore of 4.96 was obtained at a new polymorphic marker D1S21401. Haplotype analysis defined a 16 Mb critical region between D1S2868 and afmb014zb9. The interval of DFNB32 locus overlap with DFNA37 locus and the Marshall and Stickler syndromes locus. The entire coding region of COL11A1, responsible of the later syndromes, was screened and no mutation was observed. Towards the identification of the DFNB32 gene, a search on the Human Cochlear cDNA Library and EST Database was done. The genes corresponding to the ESTs found in the DFNB32 interval are being screened for deafness-causing mutations. PMID- 12634868 TI - A candidate region for Asperger syndrome defined by two 17p breakpoints. AB - Asperger syndrome (AS) is a mild form of autistic disorder characterised by impairment in social interaction as well as a restricted pattern of behaviour, interests, and activities. Two patients with AS and balanced translocations t(13;17) and t(17;19), respectively, were identified. Fluorescent in situ hybridisation (FISH) analysis with chromosome 17 specific clones to metaphase chromosomes from both patients showed that the chromosome 17 breakpoints are located within a 300 kb region at 17p13. The region spans 14 known genes. The expression of these genes was analysed in lymphoblastoid RNA derived from the patients and healthy control individuals. The CHRNE, DKFZP566H073, LOC90048, PFN1, SPAG7, KIAA0909, ZNF232 and KIF1C genes showed similar levels of expression in cell lines with the translocations when compared with cell lines with normal karyotype. No expression was detected for the MINK, GP1BA, SLC25A11, ENO3, FLJ10060 and USP6 genes in any of the cell lines. The close physical relation of the two 17p breakpoints suggest a common genetic aetiology for the phenotype in the patients. Structural and functional analysis of the genes located around the two 17p breakpoints in t(13;17) and t(17;19) patients may reveal candidate sequences for the AS phenotype. PMID- 12634869 TI - Carrier frequency of the V377I (1129G>A) MVK mutation, associated with Hyper-IgD and periodic fever syndrome, in the Netherlands. AB - Hyper-IgD and periodic fever syndrome (HIDS) and mevalonic aciduria (MA) are two autosomal recessive disorders that both are caused by a deficient activity of the enzyme mevalonate kinase (MK) due to mutations in the encoding gene (MVK). The most frequently occurring MVK mutation, V377I (1129G>A), has been identified exclusively in HIDS patients. Other common mutations have been associated with both HIDS and MA. To estimate the incidence of MK deficiency in the Netherlands, we determined the carrier frequency of the V377I mutation in genomic DNA extracted from anonymised newborn screening cards by PCR-RFLP. We found 14 carriers among 2138 analysed samples (1 : 153). Based on the V377I allele frequency of 42% in patients diagnosed with MK deficiency, the carrier frequency of any MVK mutation in the Dutch population can be calculated as 1 : 65. This predicts a disease incidence between 1 in 5196 and 1 in 53 656, which is far more than actually observed. Although under-diagnosis of patients with MK deficiency remains possible, this discrepancy probably is due to a reduced penetrance of V377I homozygosity. Analysis of the distribution of the V377I allele within patients carrying MVK mutations revealed that this was not according to the Hardy Weinberg equilibrium principle, most probably due to an under-representation of V377I homozygotes in HIDS. Homozygotes for V377I might exhibit a much milder phenotype of MK deficiency or no disease-phenotype at all. PMID- 12634870 TI - Spectrum of mutations in PTPN11 and genotype-phenotype correlation in 96 patients with Noonan syndrome and five patients with cardio-facio-cutaneous syndrome. AB - Noonan syndrome (NS) is a relatively common, but genetically heterogeneous autosomal dominant malformation syndrome. Characteristic features are proportionate short stature, dysmorphic face, and congenital heart defects. Only recently, a gene involved in NS could be identified. It encodes the non-receptor protein tyrosine phosphatase SHP-2, which is an important molecule in several intracellular signal transduction pathways that control diverse developmental processes, most importantly cardiac semilunar valvulogenesis. We have screened this gene for mutations in 96 familial and sporadic, well-characterised NS patients and identified 15 different missense mutations in a total of 32 patients (33%), including 23 index patients. Most changes clustered in one exon which encodes parts of the N-SH2 domain. Five of the mutations were recurrent. Interestingly, no mutations in the PTPN11 gene were detected in five additional patients with cardio-facio-cutaneous (CFC) syndrome, which shows clinical similarities to NS. PMID- 12634871 TI - Endothelial dysfunction in diabetes mellitus: role in cardiovascular disease. AB - Diabetes mellitus is associated with an increased risk of cardiovascular disease (CVD), even in the presence of intensive glycemic control. Substantial clinical and experimental evidence suggests that both diabetes and insulin resistance cause a combination of endothelial dysfunctions, which may diminish the anti atherogenic role of the vascular endothelium. Endothelial dysfunctions that have been described include decreased endothelium-dependent vasorelaxation, increased leukocyte-endothelial cell adhesion and vascular permeability, and the altered production of a variety of vasoactive substances, which affect coagulation, extracellular matrix homeostasis, and smooth muscle physiology. The primary mechanisms that contribute to these endothelial dysfunctions in diabetes appear to involve the activation of protein kinase C (PKC) pathways, increased non enzymatic glycation, increased oxidant stress, and reduced endothelial insulin action. In addition, many of the adverse effects of these abnormalities associated with hyperglycemia and insulin resistance are mediated and amplified by potent vasoactive hormones including angiotensin II, transforming growth factor-beta, and vascular endothelial growth factor. Multiple interventions have been shown to improve endothelial dysfunction in diabetes, including PKC inhibition, infusion of soluble receptors for advanced glycation end-products, antioxidant and insulin supplementation, and angiotensin-converting enzyme inhibition. These findings are consistent with a model involving a combination of factors contributing to the etiology of the endothelial dysfunctions in diabetes. Further work is needed to determine whether endothelial function can be used as a therapeutic target to reduce CVD and improve clinical outcomes. PMID- 12634872 TI - Systolic and diastolic heart failure--diagnostic and therapeutic dilemmas. AB - Clinical suspicion of congestive heart failure (CHF) always requires a careful diagnostic workup. This comprises the verification of the presence of CHF (in contrast to other conditions that cause nonspecific phenomena such as shortness of breath and edema), evaluation of the underlying cause of heart failure, and assessment of left ventricular (LV) systolic function. In addition to clinical examination, echocardiography is warranted in most cases. On the basis of this information, patients can be selected for further studies, such as exercise testing, cardiac catheterization and coronary angiography. In view of the serious prognosis of heart failure, especially systolic CHF, the threshold for specialist consultation should be low. Although the classification of CHF into systolic and diastolic forms is complex, clinically meaningful data can be derived simply by determining whether LV systolic function is impaired (predominantly systolic CHF) or not (probable diastolic CHF). In the latter case, treatment is mainly symptomatic in addition to the management of the underlying condition (e.g. hypertension). In systolic CHF, considerable therapeutic advances have recently been made and it is important that patients receive appropriate care to improve their prognosis. These measures include angiotensin-converting enzyme inhibitors, beta-blockers and spironolactone. PMID- 12634873 TI - Chronic heart failure as a metabolic disorder. AB - Congestive chronic heart failure (CHF) is a progressive disorder in which a complex interaction of haemodynamic, neurohormonal and metabolic disturbances leads to subsequent immune activation. The greatest attention has been given to the concept that the progression of heart failure is due to neurohormonal abnormalities and this has led to substantial therapeutic benefits for CHF. The aim of this review is to describe a number of the interactions between neurohormonal pathways and metabolic problems relevant in CHF. Besides the renin angiotensin-aldosterone-system, steroid and thyroid hormones, growth factors, insulin and inflammatory cytokines (e.g. tumour necrosis factor-alpha [TNF alpha]) are considered. TNF-alpha is potentially a key molecule with enormous interactive opportunities within a regulatory network of energy metabolism, immune function and neuroendocrine and hormonal function. The most dramatic metabolic problem in heart failure patients is the development of cardiac cachexia. Currently, no specific therapy exists and the prognosis is poor. There are promising approaches (counteracting TNF-alpha or applying anabolic growth factors) but these are not without risk and are expensive, and their application may, therefore, be limited to certain subgroups of patients. In the future, it will not be enough to monitor cardiac function and symptomatic status in heart failure patients. Rather, the patients' metabolic status may need to be taken, as well as an assessment of peak oxygen consumption, body composition and hormonal status. PMID- 12634874 TI - Cytokines and heart failure. AB - In many forms of cardiomyopathic left ventricular (LV) dysfunction, there is a rapid myocardial expression of pro-inflammatory cytokines such as interleukin 1, interleukin 6 and tumour necrosis factor-alpha (TNF-alpha) which mediate, via specific receptors, various processes such as gene expression, cell growth or apoptosis. In the initial stages of myocarditis, the myocardial expression of proinflammatory cytokines appears to be part of an inflammatory process. In many other conditions such as ischaemic cardiomyopathy and chronic LV pressure or volume overload, myocardial expression of proinflammatory cytokines is triggered by an elevation of LV wall stress. Myocardial expression of cytokines contributes to depression of contractile performance and adverse LV remodelling. Cytokine induced depression of contractile performance appears to result from sphingosine production, which interferes with myocardial calcium handling. In transgenic mice, the rate of progression of LV dilatation appears to correlate with the intensity of myocardial TNF-alpha overexpression. In heart failure patients, cytokine concentrations are elevated not only in the myocardium but also in plasma. Cytokines are, therefore, responsible not only for autocrine and paracrine signalling within the myocardium but also for endocrine signalling throughout the body, especially affecting striated muscle mass with induction of muscle wasting and cachexia. The source of cytokine production in heart failure remains uncertain and several mechanisms have been proposed including endotoxin induced immune activation due to bowel oedema, myocardial production due to haemodynamic overload and peripheral extramyocardial production due to tissue hypoperfusion and hypoxia. The latter seems to be the most likely mechanism, possibly modulated by the presence of bacterial endotoxins released from the gut. Numerous drugs have meanwhile been shown to influence this cardioinflammatory response to heart failure either by reducing basal levels of cytokines (e.g. amlodipine, pentoxifylline, beta-blockers) or by reducing endotoxin-induced cytokine gene expression (e.g. ouabain, amiodarone, adenosine, angiotensin converting enzyme inhibitors, angiotensin II-receptor blockers). Direct blockade of the deleterious actions of elevated plasma levels of cytokines recently became possible through intravenous infusion of a soluble TNF-alpha receptor fusion protein, which resulted in an increase in exercise tolerance and LV performance. PMID- 12634875 TI - beta-Blocker therapy in heart failure. AB - Heart failure is an important public health problem and one for which morbidity and mortality remain high despite treatment with angiotensin converting enzyme (ACE) inhibitors. A large number of clinical trials examining the effects of beta blockers in the treatment of heart failure have now been performed. Two large scale clinical trials have recently confirmed significant survival benefits with these agents, with effects that are additive to those achieved with ACE inhibitor therapy. These trials have now established beta-blocker therapy as an important part of standard heart failure treatment. The clinical use of beta-blockers in patients with heart failure requires careful translation of the randomized controlled trials into everyday clinical practice. Patient selection is key to the safe use of beta-blockers. Patients who may be suitable for beta-blockade therapy include those with mild-moderate heart failure due to left ventricular systolic impairment, those who are receiving adequate dose of diuretics and ACE inhibitors and those whose clinical condition is stable at the time of initiation of the beta-blocker. Survival benefits have been demonstrated with bisoprolol, carvedilol and metoprolol. Whether different beta-blockers have important clinical differences with regard to clinical end-points is as yet uncertain. beta Blockers should be initiated at low dose, with titration of dose over several weeks and careful clinical monitoring for potential adverse effects, such as hypotension or worsening congestion. This careful application of the clinical trials into clinical practice will allow the safe use of this effective treatment for patients with chronic heart failure. PMID- 12634876 TI - Polypharmacy (or polytherapy) in the treatment of heart failure. AB - There is now conclusive evidence that most patients with heart failure due to left ventricular systolic dysfunction should be treated with angiotensin converting enzyme (ACE) inhibitors and beta-blockers. They will also need diuretics for the control of fluid retention. There is also a powerful case for adding spironolactone to the treatment of patients with more severe symptoms. Many doctors would also use digoxin and, especially if coronary disease is present, aspirin or warfarin. Most patients also have other chronic diseases, such as diabetes, arthritis, depression and dyspepsia, and each of these may provoke the prescription of yet another agent. Many patients will receive prescriptions to treat the side-effects of their therapy. Finding a sure path through the morass of pharmacotherapy is a daunting task. Polypharmacy is having a negative impact on new drug research in an area where there are in fact remarkably few really effective treatments and the therapeutic problem is only partially solved. This paper discusses some of the issues surrounding polypharmacy in heart failure and how to resolve them, using an illustrative case history. It highlights the potential benefits of polypharmacy with effective drugs and the gross over-use of ineffective treatments in heart failure. The major problem with polypharmacy in heart failure is not the heart failure treatment itself, but the drugs for other concomitant conditions, the effectiveness of which is often not supported by an appropriate evidence base and for which alternative, less noxious management strategies often exist. Polypharmacy may be deleterious not only because of the increased potential for side-effects and drug interactions but also because taking unnecessary therapy reduces compliance with effective drugs. PMID- 12634877 TI - Should patients with chronic heart failure be treated with "statins"? AB - There are a number of theoretical reasons as to why 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) should be prescribed to patients with chronic heart failure (CHF). These agents are proven to prevent coronary heart disease, the major etiological factor in the development of CHF. Potential additional effects of these agents include inhibition of proinflammatory cytokine activity and other potential beneficial effects on cardiac remodeling. However, there are also possible adverse effects of this strategy, supported by the overriding observation that low plasma lipid levels portend a poorer prognosis in patients with established CHF. Potential mechanisms by which statins may directly confer adverse effects include a reduction in levels of the antioxidant ubiquinone and an increase in blood endotoxin levels, both of which may contribute to CHF disease progression. Given these uncertainties, an answer to the question of whether or not therapy for CHF should include statins requires a definitive clinical trial. The importance of such a trial is further highlighted by the already commonplace usage of statins amongst patients with CHF. PMID- 12634878 TI - Prognosis of patients with heart failure compared with common types of cancer. AB - Heart failure (HF) is a growing epidemic that not only exerts an enormous burden on the healthcare system but also imparts extremely poor quality of life in the increasing number of patients dying from this fatal syndrome. For the majority of these patients, end-of-life care is clearly suboptimal. It is within this context that this article reviews the case for comparing the prognosis of HF with that of the most common types of cancer. It also summarizes the numerous difficulties in trying to make such comparisons and highlights key comparative data--the majority of which indicate that the prognosis of HF is comparable with that of cancer. Finally, it discusses how these comparisons can highlight the need to extend the type of public health response to HF usually reserved for high profile disease states, such as cancer. PMID- 12634879 TI - Asymptomatic left ventricular dysfunction: an overlooked part of the continuum of heart failure. AB - Over 4.7 million Americans have heart failure, and 50000 new cases of heart failure are diagnosed every year. In addition, a significant proportion of the population may have asymptomatic left ventricular (LV) systolic dysfunction. Population studies estimate that the prevalence of asymptomatic LV dysfunction ranges from 0.9-5.9%. Of these, a substantial number will eventually develop symptomatic heart failure. Diagnosing and treating these patients before they develop symptoms may help to delay or prevent the development of symptomatic heart failure. This review highlights the currently available data on the epidemiology, pathophysiology, diagnosis, and treatment of asymptomatic LV systolic dysfunction. PMID- 12634880 TI - The role of invasive electrophysiology in the management of patients with chronic heart failure. AB - It is well known that both atrial and ventricular arrhythmias play a key role in morbidity and overall mortality among patients with heart failure. In addition to pharmacological treatment, up-to-date and evidence-based use of invasive electrophysiology, including implantable cardioverter defibrillator implantation, is recommended in the global management of patients with heart failure. This article will review current clinical indications for invasive electrophysiology, either acknowledged or under evaluation, focusing on the scientific background and some technical and practical aspects. The discussion is organized in an arrhythmia-based manner so that ventricular-, atrial-, and heart transplant related arrhythmias will be discussed separately. PMID- 12634881 TI - Does growth hormone play a role in chronic heart failure? AB - Experimental and clinical studies have recently demonstrated that the growth hormone-insulin-like growth factor-I (GH-IGF-I) system is involved in the regulation of cardiac structure and function. Patients with acromegaly have an increased propensity of developing cardiovascular complications, such as ventricular hypertrophy with interstitial fibrosis. Conversely, patients with GH deficiency can exhibit ventricular dysfunction, increased vascular thickness, and an increased number of atheromatous plaques. In both groups of patients these abnormalities may be partially reverted by normalizing GH-IGF-I levels. In experimental or human chronic heart failure (CHF), GH administration increases ventricular mass and cardiac performance and reduces pulmonary vascular resistance. The mechanism by which this occurs is still unclear, but seems to involve calcium channels and non-endothelium-mediated vasodilatation. Randomized trials studying CHF patients contradict these results, highlighting that, in patients with heart failure, the response to GH therapy appears to be variable, and is probably influenced either by acquired GH resistance or by baseline levels of hormones. Due to the small number of patients examined to date, larger, randomized, controlled studies are needed. PMID- 12634882 TI - Novel mechanisms of sympatho-excitation in chronic heart failure. AB - Chronic heart failure (CHF) is associated with an increase in the production and secretion of various regulatory hormones that are initially beneficial, but become deleterious when elevated for prolonged periods. The neurohumoral excitation that occurs in the CHF state is mediated, in part, by abnormal inhibitory cardiovascular reflexes, such as the arterial baroreflex and the cardiopulmonary reflex. In addition, two sympatho-excitatory reflexes have been shown to be enhanced in CHF: the arterial chemoreflex and the cardiac sympathetic afferent reflex. While these reflexes may play a role in the sympatho-excitation of the CHF state, there is an important central modulation of sympathetic outflow by a variety of hormones that are elevated in CHF and have been shown to have neural effects. These include angiotensin II (Ang II), nitric oxide (NO), and endothelin-1. In fact, experimental animal data suggest that a central reciprocal relationship exists between Ang II and NO in their ability to modulate sympathetic outflow. These substances may also participate in the beneficial effects of exercise training in the CHF state. Exercise training lowers sympathetic nerve activity and plasma Ang II, and enhances arterial baroreflex function. This review emphasizes the neurohormonal and reflex regulation of sympathetic outflow in heart failure. While abnormal reflex regulation may predict a poor outcome, new treatment options may emerge from a better understanding of reflex regulation in CHF. PMID- 12634883 TI - The impact of obesity on survival in patients with heart failure. AB - Heart failure (HF) is an important cause of morbidity and mortality. Obesity is an increasingly prevalent condition that has been associated with increased cardiovascular risk, including increased risk of developing HF. Based on the associations of obesity with cardiac structural and hemodynamic alterations, as well as case reports of reversal of cardiomyopathy with weight loss, obesity has been presumed to have a deleterious effect in patients with HF. However, several recent studies have shown that in patients with established HF, obesity is not associated with increased mortality, but rather is associated with improved survival. Potential mechanisms for cardioprotection in obesity include a diminished activation of the neurohumoral system, an enhanced protection against endotoxin/inflammatory cytokines, and an increased nutritional and metabolic reserve. Further investigations into the relationship between obesity and the progression of HF are necessary. Ultimately, clinical trials are needed to provide definitive guidance to the management of obese and overweight HF patients. PMID- 12634884 TI - Who and how to treat with non-surgical myocardial reduction therapy in hypertrophic cardiomyopathy: long-term outcomes. AB - Iatrogenic infarction of the hypertrophic intraventricular septum in hypertrophic obstructive cardiomyopathy has become an accepted treatment for patients refractory to medical treatment and/or pacemaker therapy. During the 8 years of its existence, non-surgical myocardial reduction (NSMR), which is based on the injection of absolute alcohol into the first or second septal perforator originating from the left anterior descending coronary artery, has been used more frequently than surgical myectomy. The advantages of this catheter procedure are local anesthesia, short hospital stay, and less morbidity as compared with open heart surgery. Patients with an interventricular septal thickness of at least 18 mm, left ventricular outflow tract gradient at rest of at least 30 mmHg, and an intraventricular gradient during provocation (such as isoproterenol, dobutamine, amylnitrate, and postextrasystolic potentiation) are potential candidates for this procedure. Important mitral valve abnormalities must be excluded prior to septal ablation with alcohol. In some patients, the coronary anatomy is unsuitable for this procedure. Mid-to-long-term results have shown very significant intraventricular gradient reduction, symptom improvement, reduction in left ventricular filling pressure and pulmonary artery pressure, and increase in exercise capacity. Complications, such as the need for long-term pacing, have fallen with the reduction of the total amount of injected alcohol and the use of contrast echocardiography. NSMR appears to be effective, and can be used as an alternative to classical surgical myectomy in symptomatic patients resistant to conservative treatment. PMID- 12634885 TI - Antiarrhythmic therapy in heart failure. AB - Heart failure is the term used for a cardiovascular syndrome whose definition lacks uniform criteria. It is associated with a very high mortality rate. Approximately 50% of deaths in patients with heart failure are sudden, mostly due to ventricular tachycardia (VT). In severe heart failure, death may also occur due to bradyarrhythmias. Other arrhythmias complicating heart failure include atrial and ventricular extrasystoles, atrial fibrillation, and sustained or non sustained VT. Depending on the etiology of heart failure, different preconditions, including ischemia or structural alterations (such as fibrosis) may be prominent. Re-entrant mechanisms around scar tissue, afterdepolarizations, and triggered activity due to changes in calcium metabolism significantly contribute to arrhythmogenesis. The treatment of the underlying disease process and optimal management of heart failure is of major importance. Revascularization, beta-blocker therapy, and angiotensin converting enzyme inhibitors are all essential to appropriate therapy. Treatment of arrhythmias is performed either because patients are symptomatic or to reduce the risk of sudden cardiac death. The implantable cardioverter-defibrillator (ICD) is the best available therapy to prevent sudden cardiac death from VT. Devices with back-up pacing also offer protection against bradyarrhythmias. There is evidence that patients with sustained VT or a history of resuscitation have the best outcome with ICD therapy regardless of the degree of heart failure. Many of these patients require additional antiarrhythmic therapy (e.g. amiodarone) because of atrial fibrillation or non-sustained VT that may activate the device. PMID- 12634886 TI - Mechanical circulatory support devices--state of the art. AB - Mechanical circulatory support devices (MCSDs) have been developed with the aim of bridging patients with advanced heart failure to cardiac transplantation, to recovery, and to serve as permanent support devices. The current generation of devices provides a differentiated spectrum of support, ranging from short-term to intermediate- and long-term duration, partial left ventricular (LV), complete LV, right ventricular, and biventricular options which can be tailored individually. The device positions range from paracorporeal to intracorporeal pumps with transcutaneous drivelines, to completely implantable systems. Major limitations are infection, coagulopathies, and device dysfunction. In this review, we discuss the history and experience with currently available MCSD options capable of supporting the circulation for 30 days. PMID- 12634887 TI - The development of new medical treatments for acute decompensated heart failure. AB - There were almost 1 million hospitalizations for heart failure in 1999, representing a 155% increase over the last 20 years, and the treatment of these patients is an important and growing problem. However, currently available therapies, which are based on three basic mechanisms of action (diuresis, exogenous vasodilators, and cyclic adenosine monophosphate-dependent positive inotropes), have significant limitations that have encouraged the development of newer agents. The leading medications for this indication are representatives of three different therapeutic approaches, which include endogenous vasodilatory neurohormones (nesiritide), calcium sensitizers (levosimendan), and neurohormonal antagonists (tezosentan). These three agents represent a new generation of therapeutics for this important medical problem and may provide the means not only to treat symptoms, but also to improve longer-term clinical outcomes. PMID- 12634889 TI - Are angiotensin II receptor antagonists indicated in chronic heart failure? AB - Angiotensin II type I receptor antagonists (ARAs) have been tested in two large randomized trials, Evaluation of Losartan in the Elderly II (ELITE II) and the Valsartan Heart Failure Trial (Val-HeFT), and several other large trials are ongoing in the indication of chronic heart failure (CHF). Based on the available evidence, angiotensin converting enzyme inhibitors remain the cornerstone in the treatment of CHF. However, much more information should be available in the next few years, which will provide more evidence on how to use ARAs and in which patients. PMID- 12634888 TI - Renal function as a predictor of prognosis in chronic heart failure. AB - The incidence of chronic heart failure (CHF) has been increasing, particularly because of the aging of the population and the improved survival of patients with coronary artery disease. Therefore, the current pathophysiological and clinical considerations in the diagnosis and treatment of CHF will need further improvement in terms of cardiovascular risk profiling, preventive measures, earlier intervention, and patient-tailored disease management. To date, the role of the kidney in CHF is mainly considered within the context of excessive salt and water retention, due to reduced renal blood flow. However, recent data indicate that the kidney may play a more decisive role in the progression and prognosis of the disease. It has been demonstrated that renal function is independently associated with an increased risk for all-cause mortality and cardiovascular morbidity. Furthermore, moderate renal insufficiency is a common phenomenon in this patient population and, for example, left ventricular ejection fraction, glomerular filtration rate, and New York Health Association class are not only prognostically important but are also acting independently, and support the hypothesis that cardiac function, clinical status, and renal function represent, in part, different prognostic entities of CHF. It could be questioned why an impaired renal function adds prognostic risk to develop CHF? A subclinically decreased renal function is unlikely to be the direct cause. Renal function is known to correlate with a variety of cardiovascular risk factors. Similar risk factors could contribute to the pathogenesis of intrarenal disease. Furthermore, a large number of metabolic abnormalities are related to impaired renal function and induce myocardial dysfunction and damage. Finally, neurohormonal activation is apparent in patients with chronic heart failure. Angiotensin II, the central product of the renin-angiotensin system, may play a central role in the pathophysiology and progression of cardiovascular and renal diseases. In conclusion, to prevent cardiovascular morbidity and mortality, new therapeutic strategies might be triggered by focussing on increasing our knowledge concerning adaptive and maladaptive mechanisms of the kidney involved in CHF. PMID- 12634890 TI - The importance of tumor necrosis factor and lipoproteins in the pathogenesis of chronic heart failure. AB - Elevated levels of proinflammatory cytokines, such as tumor necrosis factor (TNF) and interleukin-6 (IL-6), have been demonstrated in patients with chronic heart failure (CHF). Evidence suggests that cytokines such as these may play a central role in the pathogenesis of this syndrome. TNF has several properties that are particularly detrimental in CHF, such as negatively inotropic effects, the promotion of left ventricular remodelling, and the induction of dilated cardiomyopathy in humans. Furthermore, TNF can cause skeletal muscle wasting and apoptosis, and, therefore, may be important in the development of cardiac cachexia. Although the precise stimulus for immune activation in CHF is unknown, one hypothesis is that endotoxin may be a significant trigger for cytokine release. This is supported by the finding that decompensated CHF patients have elevated endotoxin levels that normalize on diuretic therapy. The factors that influence endotoxin responsiveness in patients with CHF, in particular the potential importance of serum lipoproteins, will be discussed in this review. PMID- 12634891 TI - Biventricular pacing in patients with heart failure and intraventricular conduction delay. AB - In patients with advanced chronic heart failure, characterized by prolonged QRS duration and by decreased cardiac contractility, decreasing dysynchrony by biventricular pacing seems to improve exercise tolerance (6-min walk distance), symptoms (New York Health Association class), and quality of-life scores. Although the results of several reports were consistent, the numbers of patients studied were small, and many of the changes were trends that did not reach statistical significance. The availability of a non-pharmacological treatment that improves exercise capacity and quality-of-life would be a major advance. However, further studies will need to address the question of mortality and morbidity benefits of such intervention. PMID- 12634892 TI - Vasopeptidase inhibitors: potential role in the treatment of heart failure. AB - Current thinking views the progression of heart failure as the result of sustained activation of vasoconstrictor neurohormones. In this model, the sustained synthesis of vasoconstrictor neurohormones leads to disease progression through alterations in cardiomyocyte structure and function, which affects myocardial contractility, cardiac metabolism, and cellular growth. Ultimately, these events induce irreversible adverse ventricular remodeling through myocyte cell loss and progressive myocardial fibrosis. In the past decade, several landmark clinical trials tested the neurohormonal hypothesis, by targeting the activation of both the beta-adrenergic and the renin-angiotensin-aldosterone systems. Although the observed decrease in mortality using this strategy in heart failure populations was encouraging, morbidity and mortality levels remained elevated, and it has now been shown that several other humoral interactions are at play and potentially deserve antagonizing, or in the case of vasodilator neurohormones, deserve stimulation. It is known a family of vasodilator neurohormones - the natriuretic peptides - that have natriuretic, vasodilatory, and antiproliferative effects, endogenously inhibit the renin-angiotensin system. These peptides are degraded primarily by a neutral endopeptidase (NEP), an endothelial cell-surface zinc metallopeptidase, which shares a similar structure and catalytic site with the angiotensin converting enzyme (ACE). NEPs have broad substrate specificity, encompassing atrial natriuretic peptide, brain natriuretic peptide, and C-type natriuretic peptide, but also bradykinin and adrenomedullin. The recognition that ACE and NEP enzymes had related structures, led to the design and development of a class of molecules with a dual inhibitory effect on ACE and NEP, referred to as vasopeptidase inhibitors. Preliminary clinical trials in heart failure with vasopeptidase inhibitors have become available and show promising results. Thus, the combined inhibition of ACE and NEP, by attenuating excessive vasoconstriction and enhancing vasodilator substances, holds promise as a valuable option in heart failure treatment for the near future. PMID- 12634893 TI - The cytokine network in heart failure: pathogenetic importance and potential therapeutic targets. AB - Accumulating evidence indicates that inflammatory mediators are important in the pathogenesis of chronic heart failure. Several studies have shown raised levels of inflammatory cytokines in patients with congestive heart failure (CHF), in both plasma and circulating leukocytes, as well as in the failing myocardium itself. Importantly, many of the inflammatory cytokines (e.g. tumor necrosis factor-a and interleukin-6) have the potential to negatively influence heart contractility, induce hypertrophy, and promote apoptosis or fibrosis, thereby contributing to the continuous remodeling process in CHF. Traditional cardiovascular drugs seem to have little influence on the cytokine network in CHF patients, and immunomodulatory therapy, in addition to 'optimal' cardiovascular treatment regimens, has emerged as an option. Thus, several small studies with therapy targeted against inflammatory mediators have shown promising effects on functional capacity and myocardial performance. These studies suggest a potential for immunomodulating therapy, in addition to optimal conventional cardiovascular treatment regimens in CHF patients. However, the results in these small studies will have to be confirmed in larger placebo-controlled mortality studies. More importantly, further research in this area will have to precisely identify the most important components in the immunopathogenesis of chronic heart failure, in order to develop more specific immunomodulating agents in this disorder. PMID- 12634894 TI - Diabetes and dyslipidemia. AB - Numerous prospective cohort studies have indicated that diabetes mellitus (DM), particularly type-2 DM (the type of diabetes associated with insulin resistance that usually strikes adults), is associated with a 3-4-fold increase in risk for coronary heart disease (CHD) [1-3]. The increase in risk is particularly evident in younger-age groups, and in women: females with type-2 DM appear to lose a great deal of the protection that characterizes non-diabetic females. Furthermore, patients with DM have a 50% greater in-hospital mortality, and a 2 fold increased rate of death within 2 years of surviving a myocardial infarction. Overall, CHD is the leading cause of death in individuals with DM who are >35 years old. Although a significant portion of this increased risk is associated with the presence of well-characterized risk factors for CHD, a significant proportion remains unexplained. Patients with DM, particularly those with type-2 DM, have abnormal plasma lipid and lipoprotein concentrations that are less commonly present in non-diabetics [4-6]. Patients with poorly controlled type-1 DM can also have a dyslipidemic pattern, but, in this review, we will focus on the dyslipidemia seen commonly in patients with type-2 DM. In particular, we will describe the pathophysiology underlying the increase in plasma very low-density lipoprotein triglyceride levels, the reductions in plasma high-density lipoprotein cholesterol levels, and the abnormal, small, dense low-density lipoproteins that are the central components of diabetic dyslipidemia. The dyslipidemia of DM clearly adds significantly to the high risk for CHD in this group, and must be treated aggressively with diet, weight loss and lipid-altering medications. Combinations of lipid-altering medications, particularly statins and fibrates, can markedly change plasma lipid levels, often bringing them all into the normal range. PMID- 12634895 TI - Experimental options in the treatment of heart failure: the role of cytokine antagonism. AB - Recent studies have identified the importance of biologically active molecules, such as neurohormones, as mediators of disease progression in heart failure. More recently, it has become apparent that, in addition to neurohormones, another portfolio of biologically active molecules, termed cytokines, are also expressed in the setting of heart failure. This article will review recent clinical material that suggests that tumor necrosis factor, a pro-inflammatory cytokine, may contribute to disease progression in heart failure by virtue of the direct toxic effects that this molecule exerts on the heart and circulation. In addition, this article reviews the existing clinical literature, which suggests that cytokine antagonism is safe and potentially effective in patients with heart failure. PMID- 12634896 TI - Ischemic versus non-ischemic heart failure: should the etiology be determined? AB - In epidemiological surveys and in large-scale therapeutic trials, the prognosis of patients with ischemic heart failure is worse than in patients with a non ischemic etiology. Even heart transplant candidates may respond better to intensified therapy if they have non-ischemic heart failure. The term 'non ischemic heart failure' includes various subgroups such as hypertensive heart disease, myocarditis, alcoholic cardiomyopathy and cardiac dysfunction due to rapid atrial fibrillation. Some of these causes are reversible. The therapeutic effect of essential drugs such as angiotensin-converting enzyme inhibitors, beta blockers and diuretics does not, in general, significantly differ between ischemic and non-ischemic heart failure. However, in some trials, response to certain drugs (digoxin, tumor necrosis factor-alpha, inhibition with pentoxifylline, growth hormone and amiodarone) was found to be better in non ischemic patients. Patients with ischemic heart failure and non-contracting ischemic viable myocardium may, on the other hand, considerably improve following revascularization. In view of prognostic and possible therapeutic differences, the etiology of heart failure should be determined routinely in all patients. PMID- 12634897 TI - Chemosensitivity in chronic heart failure. AB - Augmented peripheral and central chemoreceptor sensitivity has recently been demonstrated in both experimental and clinical settings of chronic heart failure (CHF). As a result of the effects of chemoreflexes on the respiratory, circulatory and neurohormonal systems, changes in their activity may account for several pathophysiological features of CHF--predominantly augmented ventilation, abnormal cyclic respiratory pattern and sympathetic overactivity. Although the precise underlying mechanisms are not known, a heightened chemoreflex drive may constitute an ominous sign in CHF. Patients with abnormally elevated chemosensitivity demonstrate an augmented ventilatory response to exercise, a severely impaired autonomic regulation and suppression of baroreceptor function, and a higher prevalence of ventricular arrhythmias. All these factors may unfavorably influence the prognosis of CHF. In fact, we have recently confirmed in a group of patients with advanced CHF that high peripheral chemosensitivity is an independent predictor of death. New therapies are needed in CHF to improve prognosis and quality of life. Drugs, such as opiates, and oxygen administration have been shown to suppress chemosensitivity, which may further favorably influence exercise tolerance and modify periodic breathing in CHF patients. Treatment strategies targeted at peripheral and central chemoreceptors may be a promising option for further evaluation. PMID- 12634898 TI - Elder abuse: is every bruise a sign of abuse? AB - Each year, between 1 and 2 million older adults in the United States are abused. Physicians are in an ideal position to recognize, manage and prevent elder mistreatment. This article uses a problem-solving format to discuss a case report. This case, which spanned two years, highlights the true difficulty in recognizing and managing elder mistreatment. PMID- 12634899 TI - Overview of geriatric emergencies. AB - The elderly, aged 65 or older, are more susceptible to illness or injury due to environmental factors, physiologic deterioration and co-morbid disease. Prompt referral of patients with acute decompensation to the Emergency Department promotes resuscitation, rapid evaluation and interventions where appropriate, to improve outcomes. Common life- or limb-threatening situations are reviewed as well as co-morbidities and social issues that may complicate decision making. PMID- 12634900 TI - Stability of preferences regarding life-sustaining treatment: a two-year prospective study of nursing home residents. AB - BACKGROUND: The use of advance directives is based on the consensus that physicians should respect preferences expressed by competent patients about future treatments. Patient preferences are, however, subject to change and may be influenced by a number of factors. The purpose of our study was to evaluate the durability over time of decisions made regarding terminal care of mentally intact nursing home patients and the influence of such factors as intervening illness, loss of significant others, and cognitive, emotional and functional decline. METHODS: We undertook a longitudinal prospective cohort study in which 65 mentally competent nursing home patients were interviewed at three intervals (at baseline and after one and two years). For each patient, demographic, medical and socioeconomic data were collected and assessment of mood, function, cognition and preference for life-sustaining therapies (including cardiopulmonary resuscitation and parenteral and enteral nutrition) was obtained. RESULTS: Preferences regarding cardiopulmonary resuscitation and parenteral and enteral nutrition changed over both the 12- and 24-month study periods. Only degree of change in cognitive status proved to be predictive of changes in decision. Gender, presence or absence of depression, change in level of functional abilities and intercurrent illness or stressor did not influence change regarding life sustaining therapy. CONCLUSIONS: In light of our findings, we suggest that periodic re-evaluation of these advance directives be performed and that ongoing discussions be initiated with their patients by health care professionals. PMID- 12634901 TI - Neurotrophins, growth-factor-regulated genes and the control of energy balance. AB - Neurotrophic growth factors are proteins that control neuronal differentiation and survival, and consequently play important roles in the developing and adult stages of the nervous system. Study of the genes that are regulated by these growth factors has provided insight into the proteins that are critical to the maturation of the nervous system, suggesting that select neurotrophins may play a role in the control of body homeostasis by the brain and peripheral nervous system. Our understanding of the mechanisms of action of neurotrophic growth factors has increased through experimental manipulation of cultured neurons and neuronal cell lines. In particular, the PC12 pheochromocytoma cell line, which displays many properties of adrenal chromaffin cells and undergoes differentiation into sympathetic neuron-like cells when treated with nerve growth factor, has been extensively investigated to identify components of neurotrophin signaling pathways as well as the genes that they regulate. VGF was one of the first neurotrophin-regulated clones identified in NGF-treated PC12 cells. Subsequent studies indicate that the vgf gene is regulated in vivo in the nervous system by neurotrophins, by electrical activity, in response to injury or seizure, and by feeding and the circadian clock. The vgf gene encodes a polypeptide rich in paired basic amino acids; this polypeptide is differentially processed in neuronal and neuroendocrine cells and is released via the regulated secretory pathway. Generation and analysis of knockout mice that fail to synthesize VGF indicate that this protein plays a critical, non-redundant role in the regulation of energy homeostasis, providing a possible link between neurotrophin function in the nervous system and the peripheral control of feeding and metabolic activity. Future experiments should clarify the sites and mechanisms of action of this neurotrophin-regulated neuronal and neuroendocrine protein. PMID- 12634902 TI - Behavioral cardiology --has its time finally arrived? AB - Traditional cardiology has taken a mechanistic approach to heart disease. But the new discipline of behavioral cardiology takes a broader view, concluding that heart disease is not inevitable, but develops largely from unhealthy lifestyles, such as smoking, overeating and physical inactivity, and from psychosocial stress. Physical inactivity and excessive caloric intake are also responsible for the epidemic of obesity, which is associated with a dramatic increase in the prevalence of diabetes. This increase in the incidence of diabetes may, in turn, reverse the recent decline of cardiovascular deaths in the US. A variety of psychosocial stressors have been implicated in the development of cardiovascular disease. These include occupational stress, anxiety, social isolation, hostility, anger, and type A behavior. There is clearly some overlap between these stressors, all of which may affect the heart adversely. Both the lifestyle and psychosocial factors can be altered by behavioral treatment, in which the patient and the practitioner work together. Unfortunately, various barriers can impair the successful implementation of behavioral treatment. These barriers include poor compliance by the patient, lack of skill in providing effective interventions by the health care provider, and lack of incentives within the health care system, particularly reimbursement. PMID- 12634903 TI - Hypersensitivity reactions to drugs: evaluation and management. AB - Most hypersensitivity reactions to drugs occur within several weeks of administration; signs and symptoms are often consistent with known immune mediated reactions, including anaphylaxis, rashes, fever, cytopenias and vasculitis. The culprit immune mechanisms range from immunoglobulin E antibody to T cells inducing apoptosis of keratinocytes, in the case of bullous exfoliative rashes. Many drugs induce reactions via altered hepatic metabolism, with production of reactive intermediates which induce a common syndrome of rash and fever plus variable types of other signs. Examples of this reactive metabolite syndrome include the rash and fever in HIV-positive patients given sulfamethoxazole and reactions to the aromatic anticonvulsants. With the notable exception of anaphylaxis and severe bullous exfoliative rashes, most immune reactions to drugs are not life-threatening and generally resolve once the drug is discontinued. The key is prevention. Specific immune testing is standardized only for penicillin. If test results are negative, however, the patient can tolerate all beta-lactam antibiotics. Of those patients with a positive penicillin skin test, only 2% develop reactions when given cephalosporins. Sulfa and quinolone antibiotics, and muscle relaxants, also frequently induce reactions. If there is a history of bullous rash, the patient should never again receive sulfa or quinolone, or related drugs. In other cases, a cautious graded challenge or desensitization can be done. Vancomycin, protamine, and radiocontrast media induce non-immune reactions secondary to their irritant effects on vascular endothelium. Narcotic pain medications cause histamine release by binding to a specific receptor on mast cells in sensitive patients. In contrast to true immune reactions, most patients can receive these medications again, if they are pretreated and the drugs are given slowly. Angiotensin converting enzymes, aspirin, and non-steroidal anti-inflammatory drugs induce adverse reactions by their effect on enzymes. Readministration usually results in repeat symptoms. It is possible to desensitize patients to aspirin. Some patients appear to develop similar adverse symptoms with multiple unrelated drugs. Although these cases present management problems, most patients can complete a therapeutic course of a vital drug, after careful review of the history, immune testing when possible, and graded challenge or desensitization. PMID- 12634904 TI - Meconium enhances the growth of perinatal bacterial pathogens. AB - OBJECTIVE: To demonstrate the effects of meconium on growth of bacterial pathogens, which are common causes of intra-amniotic infection and neonatal sepsis. METHODS: Meconium collected from 9 healthy neonates was suspended as a 20% solution using sterile saline. In experiment 1, separate test tubes of meconium solution and sterile saline (the control) were individually inoculated with 10(6) colony-forming units of a single species of the following test pathogens: Escherichia coli, Enterococcus faecalis, Group B Streptococcus, Staphylococcus aureus, Pseudomonas aeruginosa, and Listeria monocytogenes. After incubation at 37 degree C for 24 hours, 1 L each of the bacterial-meconium and bacterial-saline solutions was inoculated onto 5% sheep blood agar. After 24 hours of incubation, the number of developing colonies was counted. In experiment 2, equal volumes of meconium and saline solutions were inoculated with 10(5) colony-forming units of either E. coli or Group B Streptococcus. At intervals of 6, 9, and 24 hours post-incubation, 1 L each of the bacterial-meconium and bacterial-saline solutions was inoculated onto 5% sheep blood agar plates, and colonies were counted after overnight incubation. RESULTS: In the first experiment, 24 hours of incubation resulted in bacterial amplification in the meconium solution from an initial inoculum of 10(6) colony-forming units/mL to 10(9) colony-forming units/mL. In contrast, the same inoculation of saline solution (control) showed no increase in colony counts over the same time interval. For E. coli and Group B Streptococcus in experiment 2, growth enhancement in meconium was seen as early as 6 hours, as colony counts of a test species increased from 105 colony-forming units/mL to 10(9)-10(10) colony-forming units/mL. CONCLUSION: Enhanced growth of perinatal pathogens in meconium was constantly observed, and can occur as early as 6 hours after bacterial interaction of meconium. PMID- 12634905 TI - Small bowel perforation secondary to metastatic lung cancer: a case report and review of the literature. AB - Small bowel perforation is a rare complication of metastatic lung cancer. We were able to document 35 such cases in the literature in English. Aggressive investigation and early surgery is the only method for providing palliation to these patients. However, morbidity and mortality remain high and the prognosis is dire. PMID- 12634906 TI - Primary bone marrow B-cell lymphoma: report of four cases. AB - Bone marrow involvement is infrequent at presentation in cases of diffuse large B cell lymphoma. We report four adult patients with diffuse large B-cell lymphoma in whom bone marrow involvement with hematologic manifestations was the predominant clinical feature at presentation. Three patients presented with a leukoerythroblastic blood picture and one with pancytopenia. In each case, the unusual hematologic manifestations, with bone marrow replacement and the presence of immature forms in the peripheral blood, led to consideration of alternative hematologic diagnoses, including acute granulocytic leukemia in three cases and a myelodysplastic syndrome in one. The correct diagnoses were established by immunohistochemistry on formalin-fixed, paraffin-embedded bone marrow for two cases and by flow cytometry on aspirated bone marrow or peripheral blood lymphocytes for the other two. Diffuse large B-cell lymphoma should be considered in the differential diagnosis of unusual hematologic presentations, particularly in the elderly. PMID- 12634907 TI - New cytotoxic agents and molecular-targeted therapies in the treatment of metastatic breast cancer. AB - Cytotoxic chemotherapy is important for treatment of patients with hormone insensitive advanced breast cancer. A variety of new cytotoxic agents are promising alone or in combination. The originality, the clinical activity and side effects, as well as the current development status of these agents are reviewed. These agents include the new antimicrotubules (analogues of taxanes and vinorelbine; epothilone derivatives), oral formulations of 5-fluorouracil and other antimetabolites (tomudex, alimta, gemcitabine), liposomal anthracyclines, platinum analogues, topoisomerase I inhibitors and other compounds such as ET 743. Finally, new molecular-targeted therapies of potential interest in the treatment of metastatic breast cancer are reviewed. The growing availability of such biological therapies given alone and mainly in combination with hormonal and chemotherapeutic agents may improve in the near future the outcome of patients with metastatic breast cancer. PMID- 12634908 TI - Predictors of response to systemic therapy in breast cancer. AB - The most appropriate systemic therapy for a population of patients with breast cancer is determined from clinical trial data. However, the heterogeneity of breast cancer is such that within a population individual patients derive variable benefit. There is therefore a need for predictive molecular factors in order that treatment can be individualised. This review describes the roles of HER-2, epidermal growth factor receptor (EGFR), oestrogen receptor (ER)/progesterone receptor (PgR), Ki67, Bcl-2, p53 and gene expression profiling in predicting responses to endocrine, cytotoxic and biological therapies. ER and PgR remain the only well-established predictive markers of responses to endocrine therapy, although HER-2/neu has an emerging role in this area and in choice of adjuvant chemotherapy. There are considerable methodological difficulties in identifying useful predictive factors but on the basis of current evidence other biomarkers add little additional information. The development of targeted therapies means that the molecular targets themselves may become useful predictive factors for directing use of these therapies. HER-2 already has an established role in this area, but the role of EGFR requires further elaboration. The use of DNA microarrays to assess gene expression profiles may revolutionise our ability to predict responses to therapy. PMID- 12634909 TI - High-dose adjuvant chemotherapy for breast cancer: state of the art. AB - To date, seven randomised trials of high-dose chemotherapy for high-risk breast cancer have been published, both in Europe and in the US. We will not comment on the discredited Bezwoda trials. The main inclusion criteria for entry into these trials was an involvement of 4 or more axillary lymph nodes. Only two of these trials showed a trend in event-free survival favouring the high-dose arm. However, many differences in the designs of the trials were observed and general conclusions cannot be drawn as yet. The number of randomised patients spanned from 78 to 885, the scheduling and the drug combination of high-dose regimens varied notably and finally the issue of the contamination of the grafts by tumour cells still needs to be correctly addressed. We certainly are only at the end of the beginning. PMID- 12634911 TI - Chemotherapy in the elderly with breast cancer. AB - The number of elderly women diagnosed with breast cancer by 2025, will increase by 72%. These elderly women do not yet participate in most screening programmes. One notes more favourable biological characteristics of the tumour, including more expression of steroid receptors (oestrogen receptor, progesterone receptor), low proliferative rate, good differentiation, normal p53 and low expression of epidermal growth factor. Elderly breast cancer patients are frequently treated with breast conservation, omitting axillary dissection, radiation therapy and chemotherapy. There is a paucity of data to substantiate that such an approach is better than a more radical one. The efficacy of chemotherapy and guidelines for its use vary by tumour stage and patient age. Drugs like modified anthracyclines, vinorelbine, the taxanes and capecitabine may be changing the paradigm of combination therapy superiority. PMID- 12634910 TI - Interactions of antioestrogens and aromatase inhibitors. AB - Aromatase inhibitors and antioestrogens have shown substantial activity in primary and advanced breast cancer. Since they exhibit different modes of action, attempts have been made to combine them or to use them sequentially in order to potentially increase their efficacy. In preclinical studies, combined, sequential or alternating treatments with aromatase inhibitors and antioestrogens have failed to provide higher antitumoural activity. There are relevant pharmacokinetic interactions resulting in decreased plasma concentrations of third generation aromatase inhibitors when combined with tamoxifen. Several randomised clinical trials comparing single agent and combined treatment with tamoxifen and aminoglutethimide failed to show any benefit for the combination. Early results of the adjuvant ATAC trial indicate that single agent anastrozole is superior to tamoxifen or the combination of both. Several trials are ongoing which might help to further define the role of sequential or combined treatment with aromatase inhibitors and antioestrogens. However, to date, looking at the current evidence, combined treatment with aromatase inhibitors and antioestrogens does not appear to provide additional benefit compared to single agent treatment. PMID- 12634912 TI - The catalytic cycle of tyrosinase: peroxide attack on the phenolate ring followed by O[bond]O cleavage. AB - The oxidation of phenols to ortho-quinones, catalyzed by tyrosinase, has been studied using the hybrid DFT method B3LYP. Since no X-ray structure exists for tyrosinase, information from the related enzymes hemocyanin and catechol oxidase were used to set up a chemical model for the calculations. Previous studies have indicated that the direct cleavage of O(2) forming a Cu(2)(III,III) state is energetically very unlikely. The present study therefore followed another mechanism previously suggested. In this mechanism, dioxygen attacks the phenolate ring which is then followed by O[bond]O cleavage. The calculations give a reasonable barrier for the O(2) attack of only 12.3 kcal/mol, provided one of the copper ligands is able to move substantially away from its direct copper coordination. This can be achieved with six histidine ligands even if these ligands are held in their positions by the enzyme, but can also be achieved if one of the coppers only has two histidine ligands and the third ligand is water. The next step of O[bond]O cleavage has a computed barrier of 14.4 kcal/mol, in reasonable agreement with the experimental overall rate for the catalytic cycle. For the other steps of the mechanism, only a preliminary investigation was made, indicating a few problems which require future QM/MM studies. PMID- 12634913 TI - Effect of mutations in the pea genes Sym33 and Sym40. I. Arbuscular mycorrhiza formation and function. AB - Two pea (Pisum sativum L.) symbiotic mutants SGEFix(-)-1 (sym40) and SGEFix(-)-2 (sym33) with abnormalities in infection thread development and function in symbiotic root nodules have been characterised in terms of mycorrhizal colonisation of roots, shoot and root biomass accumulation and shoot and root phosphorus (P) content. The mutation in gene sym33 decreased mycorrhizal colonisation of roots (except arbuscule abundance in mycorrhizal root fragments, which increased) but did not change the effectiveness of mycorrhiza function. The mutation in sym40 did not affect either of these processes. Both mutants showed differences in plant development compared with the wild-type line SGE. The mutants had delayed flowering and pod ripening, and shoot/root biomass ratios and P accumulation also differed from those of SGE. These observations suggest that the gene mutations cause systemic changes in plant development. PMID- 12634914 TI - Effect of mutations in the pea genes Sym33 and Sym40. II. Dynamics of arbuscule development and turnover. AB - Two symbiotic pea (Pisum sativum L.) mutants SGEFix(-)-1 (sym40) and SGEFix(-)-2 (sym33) with abnormalities in infection thread formation in symbiotic root nodules were characterised with respect to dynamics of arbuscule development at 15 degrees C and 24 degrees C. Mutation of sym33 decreased mycorrhiza colonisation at both temperatures and delayed arbuscule development at 15 degrees C, whereas mutation of sym40 accelerated mycorrhiza colonisation and arbuscule senescence at 24 degrees C. The differences between the mutants and the wild-type were more pronounced at 24 degrees C, a temperature close to the optimum for pea growth. The results demonstrate that both pea genes are important in the control of arbuscular mycorrhiza development and can be considered necessary for the tripartite symbiosis in pea. PMID- 12634915 TI - The ectomycorrhizal symbiosis between Lactarius deliciosus and Pinus sylvestris in forest soil samples: symbiotic efficiency and development on roots of a rDNA internal transcribed spacer-selected isolate of L. deliciosus. AB - The effect on plant growth of pre-inoculation of Pinus sylvestris with the ectomycorrhizal (ECM) edible basidiomycete Lactarius deliciosus (isolate D45) under controlled conditions, and the development on roots of this basidiomycete, were investigated in gamma-irradiated and unsterilized containers containing different forest soil cores or a perlite-vermiculite mixture. Five months after planting, L. deliciosus mycorrhizal plants exhibited greater growth than the non mycorrhizal ones in all soil types, i.e. up to a 325% increase in shoot height in the sterilized soils. The experiment demonstrated the dependency of P. sylvestris seedlings upon ECM symbiosis for their survival in gamma-irradiated, microbiologically disturbed soil samples. Furthermore, in two soils, the growth of L. deliciosus-inoculated seedlings was greater in the sterilized soil samples than in the non-sterilized ones, i.e. 46% and 132% increase in shoot height under sterilized soil conditions. In containers randomly sampled from each soil type, the degree of root colonization by the inoculated isolate, calculated as the number of mycorrhizal root tips divided by the total number of root tips x100, ranged from 80% to 35%. Within the short term, the inoculated isolate developed rapidly on roots, dominated, and hampered ectomycorrhiza formation by various unidentified (but not Lactarius) resident ECM fungi in unsterilized soil types. Results indicate that the ECM species L. deliciosus is worth investigating to ascertain if other isolates benefit pine growth like the isolate D45, and are therefore also attractive candidates for forestry applications in the Mediterranean area. PMID- 12634916 TI - Development of microsatellite markers from an ectomycorrhizal fungus, Tricholoma matsutake, by an ISSR-suppression-PCR method. AB - An inter-simple sequence repeat (ISSR)-suppression-PCR technique established to develop microsatellite markers of plant species was applied to an ectomycorrhizal fungus, Tricholoma matsutake. Six polymorphic SSR markers were developed. All six polymorphic SSR markers were single-locused and co-dominant. Alleles produced by these six single-locused markers ranged from two to nine per locus and the expected heterozygosities were calculated as values from 0.098 to 0.803. The results indicated that the ISSR-suppression-PCR technique was effective and applicable to the development of microsatellite markers from ectomycorrhizal fungi. Furthermore, the six microsatellite loci did not amplify DNA from any other ectomycorrhizal species investigated, except for Tricholoma nauseosum (Swedish matsutake) and Tricholoma fulvocastaneum, suggesting that population genetics and reproduction of T. matsutake could be investigated by the SSR markers developed in the present study. PMID- 12634917 TI - Type of mycorrhizal associations in two coastal nature reserves of the Mediterranean basin. AB - We analysed the mycorrhizal types of two coastal Mediterranean plant communities. Plants belonging to 82 species of 46 families growing in two Mediterranean study sites located within a Regional Natural Park in Italy (Macchia Lucchese) and in a National Park in Croatia (Brijuni) were assessed for the type of their mycorrhizal associations. Mycorrhizas occurred in 83% and 90% of the plant species surveyed in Macchia Lucchese and in Brijuni, respectively. On the basis of macroscopic and microscopic characteristics, six different mycorrhizal types were found, and their co-occurrence in the same plant community showed the large mycorrhizal diversity in Mediterranean vegetation. Different occurrence patterns of mycorrhizal types have been recognised in diverse habitats within Macchia Lucchese, from sand dune plant communities to sclerophyllous woodland. Mycorrhizal type abundance calculated on a floristic basis was very different from that obtained using a vegetation cover index. PMID- 12634918 TI - In vitro evidence of mycoparasitism of the ectomycorrhizal fungus Laccaria laccata against Mucor hiemalis in the rhizosphere of Pinus sylvestris. AB - Interactions between the ectomycorrhizal fungus Laccaria laccata and the soil fungus Mucor hiemalis f. hiemalis in co-culture, and in the rhizosphere of in vitro-grown Pinus sylvestris seedlings were investigated by light- and scanning electron-microscopy. In co-culture, mycelial growth away from the L. laccata colony reduced the number of aerial hyphae at the contact zone and increased the density and compactness of the mycelium-characterized gross morphology of the saprobic fungus. Although the growth of M. hiemalis was suppressed, no penetration of M. hiemalis hyphae after the colony was entered by L. laccata was observed. Instead, dense coiling of L. laccata hyphae around sporangiophores, overpowering them and causing them to disappear, was quite common. On nonmycorrhizal roots, sporangiospores germinated heavily and formed long hyphae for 2 days post inoculation, whereas their germination was totally inhibited on mycorrhizal roots. At 3 days after inoculation, only sporangia were seen with mycelial mats firmly attached to the roots by the mantle hyphae, whereas some remnants of sporangiophores, ruptured sporangial walls and degraded hyphae of M. hiemalis were overgrown by the mantle hyphae. During the next 3 days, the mantle hyphae-invading sporangia formed short, thin branches that grew directly towards individual spores, tapering off upon contact. PMID- 12634919 TI - Changes in the concentration of trigonelline in a semi-arid leguminous plant (Prosopis laevigata) induced by an arbuscular mycorrhizal fungus during the presymbiotic phase. AB - An in vitro presymbiotic system between mesquite [Prosopis laevigata(Willd.) M.C. Johnst], a semi-arid leguminous plant, and pregerminated spores of Gigaspora rosea Nicol. & Schenck was established. After characteristic hyphal branching, high performance liquid chromatographic analyses of methanol extracts from P. laevigata roots revealed a concentration change in one ultraviolet-detectable product. This product was identified by nuclear magnetic resonance and mass spectrometry as trigonelline, a pyridine alkaloid. The concentration of trigonelline was constant in the aerial parts of the plant with or without G. rosea, but its concentration in the roots increased 1.8-fold when G. roseawas present. Trigonelline may be a regulatory factor during early signal events in the establishment of the arbuscular mycorrhizal symbiosis in P. laevigata. PMID- 12634920 TI - Variation in the FABP2 promoter alters transcriptional activity and is associated with body composition and plasma lipid levels. AB - The fatty acid-binding proteins (FABPs) are cytoplasmic proteins involved in intracellular fatty acid transport and metabolism. FABP2, the intestinal-type FABP, is expressed exclusively in enterocytes in the small intestine. In previous studies of an Ala54Thr substitution in FABP2, the Thr-allele showed association with increased lipid oxidation, elevated plasma lipids, and impaired insulin sensitivity. We screened roughly 1 kb 5' of the FABP2 initiation codon and identified three insertion/deletion polymorphisms and four single nucleotide polymorphisms (SNPs). Three of the SNPs were in complete linkage disequilibrium with the three insertion/deletion polymorphisms, defining exactly two haplotypes (FABP2p-ID). We tested the hypothesis that this variation alters gene expression by transfecting Caco-2 cells with pGL3-Basic constructs containing opposite FABP2p-ID haplotypes. Luciferase assays showed a statistically significant two fold increase in gene expression of the pGL3-insertion construct over the pGL3 deletion construct (P<0.001; n=5). We also tested for association between three FABP2 variants and measurements of body composition, plasma lipids, and insulin sensitivity in non-diabetic control subjects from the San Luis Valley Diabetes Study (n=714). The only informative variant, FABP2p-ID, was statistically significantly associated with body mass index (P=0.042) and marginally associated with fat mass (P=0.084), cholesterol (P=0.066), and HOMA IR (a derived measure of insulin resistance; P=0.062) in the entire cohort. Similar associations were seen only in non-Hispanics when the analysis was stratified by ethnicity. Within the non-Hispanic subgroup, the effects of FABP2p-ID on plasma lipids were sex specific. These results suggest that genetic variation in the 5' region of FABP2 affects transcriptional activity, presumably leading to alterations in body composition and lipid processing. PMID- 12634921 TI - System L: heteromeric exchangers of large, neutral amino acids involved in directional transport. AB - The plasma membrane transport system L is in many cells the only (efficient) pathway for the import of large branched and aromatic neutral amino acids. The corresponding transporters are hetero(di)mers composed of a catalytic subunit (LAT1 or LAT2=light chain=glycoprotein-associated amino acid transporter) associated covalently with the glycoprotein 4F2hc/CD98 (heavy chain). The tissue distribution of LAT1 suggests that it is involved mainly in transporting amino acids into growing cells and across some endothelial/epithelial secretory barriers, whereas the localization of LAT2 indicates that it is mainly involved in the basolateral efflux step of transepithelial (re)absorptive amino acid transport. However, system L transporters are obligatory amino acid exchangers with 1:1 stoichiometry, with similar (but not identical) intra- and extracellular substrate selectivities and with highly asymmetrical apparent affinities (low affinity inside). Therefore, net directional transport of large, neutral amino acids by system L depends on the parallel expression of a unidirectional transporter with overlapping selectivity (for instance systems A or N) that provides/recycles amino acids that drive system L exchange function. By mediating the regulated flux of these exchange substrates, unidirectional transporters control the activity of system L. PMID- 12634922 TI - Recruitment of renal dopamine 1 receptors requires an intact microtubulin network. AB - Renal dopamine1 receptor (D1R) can be recruited from intracellular compartments to the plasma membrane by D1R agonists and endogenous dopamine. This study examines the role of the cytoskeleton for renal D1R recruitment. The studies were performed in LLCPK-1 cells that have the capacity to form dopamine from L-dopa. In approximately 50% of the cells treated with L-dopa the D1R was found to be translocated from intracellular compartments towards the plasma membrane. Disruption of the microtubulin network by nocodazole significantly prevented translocation. In contrast, depolymerization of actin had no effect. In control cells D1R colocalized with NBD-C(6)-ceramide, a trans-Golgi fluorescent marker. This colocalization was disrupted in L-dopa-treated cells. Tetanus toxin, an inhibitor of exocytosis, prevented L-dopa-induced receptor recruitment. L-Dopa treatment resulted in activation of protein kinase C (PKC). To test the functional effect of D1R recruitment, the capacity of D1R agonists to activate PKC was studied. Activation of D1R significantly translocated PKC-alpha from intracellular compartments to the plasma membrane. Disruption of microtubules abolished D1R-mediated - but not phorbol-ester-mediated - translocation of PKC. We conclude that renal D1R recruitment requires an intact microtubulin network and occurs via Golgi-derived vesicles. These newly recruited receptors couple to the PKC signaling pathway. PMID- 12634923 TI - Calcium dependence of action potential-induced endocytosis in chromaffin cells. AB - Exocytosis occurs via fusion of transmitter-containing granules with the cell membrane, whereupon the granule contents are released and the cell membrane surface area increases. Exocytosis is followed by endocytosis to maintain proper cell membrane surface area and composition. We have shown that adrenal chromaffin cells internalize membrane in a biphasic manner following action potential stimulation. At basal firing rates (single - 0.5 Hz trains) endocytosis occurs by a rapid retrieval of membrane (termed Phase I) that is independent of the activity of the protein phosphatase calcineurin and wanes in efficiency with cell activity. At intermediate firing frequencies (>6 Hz) a second, calcineurin sensitive, form of activity-enhanced endocytosis emerges (Phase II). In this study, we employ electrophysiological, electrochemical, and computational techniques to estimate intracellular Ca(2+) at the site of endocytosis by measuring secretion rates. The measured rates of secretion yield estimates of [Ca(2+)](i) based on a kinetic scheme for exocytosis calibrated under highly controlled [Ca(2+)](i). Based on this analysis, we propose that Phase I endocytosis is inhibited by cytosolic Ca(2+) with a K(inh)=605 nM, while Phase II endocytosis is activated by Ca(2+) with a K(act)=1.46 micro M. Molecular processes that may be consistent with the measured behaviors are discussed. PMID- 12634925 TI - Cell ageing for 1 day alters both membrane elasticity and viscosity. AB - This study was designed to focus on the influences of ageing on membrane elasticity and membrane viscosity. The method used was that of postfusion red blood cell oscillation, since this does not require contact between the cells and a mechanical device to apply reproducible forces to the membrane. Freshly drawn human red blood cells were compared with cells from the same donors drawn 24 h earlier and stored at 20 degrees C. The measurements of the oscillation's first pump event time constant and the first swell phase duration revealed no significant changes between fresh and aged cells. Geometrical cell parameters alone were insufficient to characterise changes in the mechanical membrane properties, since some did not vary significantly whereas others did. On the other hand, measurement of the rates of change of geometrical parameters showed that both the membrane elasticity and viscosity modules were increased after the ageing period. Elasticity and viscosity influences could be separated because the durations of the two phases of the cell oscillations differ by three orders of magnitude. Further evidence is provided that the measurement of mechanical membrane characteristics of aged red blood cell must incorporate measurements of membrane surface properties rather than cell volume properties. PMID- 12634924 TI - Increased nitric oxide synthesis in uraemic platelets is dependent on L-arginine transport via system y(+)L. AB - Bleeding tendency in uraemic patients seems to be related to alterations in the activity of the L-arginine-nitric oxide (NO) signalling pathway in platelets. We have reported previously that L-arginine influx into human platelets is mediated by the high-affinity cationic amino acid transport system y(+)L. In the present study we examined the dependency of nitric oxide synthase (NOS) activity on L arginine transport in platelets isolated from healthy controls and uraemic patients on haemodialysis. We investigated basal and ADP-stimulated NOS activity, as reflected by the conversion of L-[(3)H]arginine to L-[(3)H]citrulline, in platelets obtained from healthy controls and uraemic patients on haemodialysis. To determine whether NOS activity depended on L-arginine transport, we analysed the effects of competitive inhibitors of L-arginine transport via system y(+)L on NOS activity. Basal NOS activity was increased from 0.21+/-0.06 to 0.7+/-0.2 pmol/10(8) platelets ( n=9, P<0.05) in uraemic patients. Stimulation by ADP (10 micro M) significantly increased NOS activity (inhibitable by L-NAME) in control platelets (252%) but failed to increase further the elevated NOS activity in uraemic platelets. Homocysteine and L-leucine, competitive inhibitors of system y(+)L, markedly inhibited NOS activity in uraemic platelets. These observations indicate that platelets from uraemic patients on haemodialysis generate more NO than control platelets and that entry of L-arginine via system y(+)L is most likely rate-limiting for platelet NO production in chronic renal failure. PMID- 12634926 TI - Phosphocreatine as a determinant of K(ATP) channel activity in pancreatic beta cells. AB - The aim of the present study was to test the hypothesis that a creatine kinase/phosphocreatine system is involved in the regulation of K(ATP) channels in pancreatic beta-cells. The phosphocreatine concentration in isolated mouse islets clearly increased in parallel with the ATP/ADP ratio in response to a rise of the glucose concentration from 0.5 mM to 15 mM. The currents through K(ATP) channels expressed in oocytes of Xenopus laevis were inhibited by injection of phosphocreatine or ATP but not by phosphate or creatine alone. In inside-out patches of beta-cell membranes obtained from native beta-cells, phosphocreatine reduced the open probability of single K(ATP) channels in the presence of ADP but not in the absence of the nucleotide. These experiments suggest the existence of a K(ATP) channel-associated creatine kinase that phosphorylates ADP. The creatine kinase inhibitor iodoacetamide suppressed the glucose-induced oscillations of the cytoplasmic Ca(2+) concentration, [Ca(2+)](c). It is concluded that phosphocreatine serves as a shuttle for energy-rich phosphate from the mitochondria to the plasma membrane. The data provide a novel model for signal transduction to K(ATP) channels in pancreatic beta-cells. PMID- 12634927 TI - UCP3 protein expression is lower in type I, IIa and IIx muscle fiber types of endurance-trained compared to untrained subjects. AB - Uncoupling protein 3 (UCP3) is a muscle mitochondrial protein believed to uncouple the respiratory chain, producing heat and reducing aerobic ATP production. Our aim was to quantify and compare the UCP3 protein levels in type I, IIa and IIx skeletal muscle fibers of endurance-trained (Tr) and healthy untrained (UTr) individuals. UCP3 protein content was quantified using Western blot and immunofluorescence. Skeletal muscle fiber type was determined by both an enzymatic ATPase stain and immunofluorescence. UCP3 protein expression measured in skeletal muscle biopsies was 46% lower ( P=0.01) in the Tr compared to the UTr group. UCP3 protein expression in the different muscle fibers was expressed as follows; IIx>IIa>I in the fibers for both groups ( P<0.0167) but was lower in all fiber types of the Tr when compared to the UTr subjects ( P<0.001). Our results show that training status did not change the skeletal muscle fiber hierarchical UCP3 protein expression in the different fiber types. However, it affected UCP3 content more in type I and type IIa than in the type IIx muscle fibers. We suggest that this decrease may be in relation to the relative improvement in the antioxidant defense systems of the skeletal muscle fibers and that it might, as a consequence, participate in the training induced improvement in mechanical efficiency. PMID- 12634928 TI - Histaminergic modulation of the intact respiratory network of adult mice. AB - Histaminergic modulation of neuronal activity in the respiratory network was investigated under normoxic and hypoxic conditions in the working heart-brainstem preparation of adult mice. Systemic application of histamine, as well as the H-1 and H-3 receptor agonists 6-[2-(4-imidazolyl)ethylamino]- N-(4 trifluoromethylphenyl) heptanecarboxamide (HTMT) and imetit, 0.5-10 micro M, significantly increased the frequency of respiratory burst discharges. Dimaprit, an H-2 receptor agonist, had no effect on respiratory activity. To test for ongoing histaminergic modulation we applied the histamine receptor antagonists pyrilamine (H-1); cimetidine (H-2) and thioperamide (H-3), each 0.5-10 micro M. Only the H-1 receptor antagonist had significant effects, viz. reduction of respiratory frequency and depression of burst amplitude. Underlying effects of histamine receptor activation were identified at the cellular level. Intracellular recordings showed that histamine mediated an increase in synaptic drive potentials in inspiratory neurones while augmentation of inhibitory and excitatory synaptic activity was observed in expiratory neurones. The augmented synaptic depolarisation of inspiratory neurones was blocked by the H-1 receptor antagonist. Histaminergic modulation is also involved in the hypoxic response of the respiratory network. Blockade of H-1 receptors significantly attenuated secondary depression of the biphasic hypoxic responses, while hypoxic augmentation was not affected. We conclude that histamine is a functional neuromodulator, which is tonically active in the respiratory network and is activated further during hypoxia. The data indicate that histaminergic neuromodulation acts predominantly via H-1 receptors. PMID- 12634929 TI - Determinants of pH sensing in the two-pore domain K(+) channels TASK-1 and -2. AB - TASK-1 and -2 are members of the two-pore domain potassium (K(+)) channel family and are sensitive to changes in extracellular pH. The effects of mutating charged, extracellular-facing residues in TASK-1 and -2 were studied in Xenopusoocytes by two-electrode voltage clamp. Hydrogen ion block was independent of voltage with K(d) values of 149+/-17.9 nM [H(+)] ( n=6) and 5.76+/-1.23 nM [H(+)] ( n=7) for TASK-1 and -2, respectively. Compared to wild-type TASK-1, H72N, H98N, H98D and K210N displayed significant shifts in their K(d) values for hydrogen ion block ([H(+)]; 110+/-9.80 nM, 737+/-170 nM, 321+/-85.9 nM and 267+/ 9.92 nM, respectively, n=6 each, P<0.05). Although significantly reducing its pH sensitivity, mutation of H98 in TASK-1 did not abolish pH sensitivity; this implies that H98 is not the only residue or domain involved in pH sensing of TASK 1. TASK-2 does not possess a histidine residue at the homologous position. However, the inclusion of such a residue failed to produce the expected increase in pH sensitivity; instead, a slight decrease was observed. Despite their structural homology and common sensitivity to pH, the TASK family of K(+) channels apparently has diverse pH-sensing mechanisms. PMID- 12634930 TI - The carboxyl terminus of the epithelial Ca(2+) channel ECaC1 is involved in Ca(2+)-dependent inactivation. AB - The family of epithelial Ca(2+) channels (ECaC) is a unique group of highly Ca(2+)-selective channels consisting of two members, ECaC1 and ECaC2. We used carboxyl terminal truncations and mutants to delineate the molecular determinants of the Ca(2+)-dependent inhibition of ECaC. To this end, rabbit ECaC1 was expressed heterologously with green fluorescent protein (GFP) in human embryonic kidney 293 (HEK293) cells using a bicistronic vector. Deletion of the last 30 amino acids of the carboxyl terminus of ECaC1 (G701X) decreased the Ca(2+) sensitivity significantly. Another critical sequence for Ca(2+)-dependent inactivation of ECaC1 was found upstream in the carboxyl terminus. Analysis of truncations at amino acid 635, 639, 646, 649 and 653 disclosed a critical sequence involved in Ca(2+)-dependent inactivation at positions 650-653. C653X showed decreased Ca(2+) sensitivity, comparable to G701X, while E649X lacked Ca(2+)-dependent inactivation. Interestingly, the number of green fluorescent cells, which is an index of the number of transfected cells, was significantly smaller for cells transfected with truncations shorter than E649 than for cells transfected with wild-type ECaC. However, the expression level of GFP was restored in the presence of the ECaC blocker ruthenium red, suggesting that these truncations resulted in deleterious Ca(2+) influx. In conclusion, we have identified two domains in the carboxyl terminus of ECaC1 that control Ca(2+) dependent inactivation. PMID- 12634931 TI - HERG K(+) currents in human prolactin-secreting adenoma cells. AB - To investigate the presence and possible function of ether-a-go-go-related gene (erg) K(+) channels in human lactotroph cells (HERG channels), primary cultures were prepared from human prolactinoma tissue. In almost all primary cultures, HERG currents could be recorded in identified prolactin cells using an external high-K(+) solution. The antiarrhythmic agent E-4031, a specific blocker of erg channels, served to isolate HERG currents as the drug-sensitive currents. In cells of two tumours tested, thyrotropin-releasing hormone significantly reduced the amplitude of the HERG currents. The potential dependence of HERG current availability and the deactivation kinetics differed significantly even between prolactin cells derived from one adenoma. For comparison, corresponding values were obtained for heterologously expressed rat erg1, erg2 and erg3 channels. The expression of the three HERG channel subunits was investigated in nine human adenomas using RT-PCR. Transcripts for HERG1 were present in all adenomas and although transcripts for HERG2 and HERG3 were also detected, their expression level was more variable. The results demonstrate the functional expression of HERG channels in human prolactin-secreting tumours and are compatible with a physiological role for these channels in the control of prolactin secretion, as has been shown in normal rat lactotroph cells. PMID- 12634933 TI - Tetrodotoxin block of A-fibre afferents from skin and muscle - a tool to study pure C-fibre effects in the spinal cord. AB - The properties of tetrodotoxin (TTX)-resistant C-fibre afferents of the dorsal roots were tested in Sprague-Dawley rats. Dorsal roots (L4-L6) were blocked with TTX (0.5-1 micro M) and the amplitude of the first response of the dorsal horn superficial interneurones (cord dorsum potential, CDP) to electrical stimulation of peripheral C-fibres in combination with natural noxious stimulation was taken as measure for intact conductivity of different kinds of noxious input by means of the C-fibre refractory period. After blockade of dorsal roots with TTX, formerly masked CDPs from muscle C-fibre afferents were uncovered. Noxious pressure to the gastrocnemius soleus muscle belly and noxious pinch to the calcanean tendon proved to be TTX resistant and therefore was propagated centrally. For cutaneous heat nociceptors it could also be shown that conductivity was intact after blockade of the dorsal roots with TTX. However, we could not exclude the TTX resistance of non-nociceptive receptors of muscle or skin. Nevertheless, blockade of afferents with TTX together with suitable stimulation techniques proves to be a reliable method to investigate central effects from C-fibre afferents without contaminating effects from A-fibres in the rat. PMID- 12634932 TI - Regulation of KCNE1-dependent K(+) current by the serum and glucocorticoid inducible kinase (SGK) isoforms. AB - The slowly activating K(+) channel subunit KCNE1 is expressed in a variety of tissues including proximal renal tubules, cardiac myocytes and stria vascularis of inner ear. The present study has been performed to explore whether the serum- and glucocorticoid-inducible kinase family members SGK1, SGK2, or SGK3 and/or protein kinase B (PKB) influence K(+) channel activity in Xenopus oocytes expressing KCNE1. cRNA encoding KCNE1 was injected with or without cRNA encoding wild-type SGK1, constitutively active (S422D)SGK1, inactive (K127 N)SGK1, wild type SGK2, wild-type SGK3 or constitutively active (T308D,S473D)PKB. In oocytes injected with KCNE1 cRNA but not in water-injected oocytes a depolarization from 80 mV to -10 mV led to the appearance of a slowly activating K(+) current. Coexpression of SGK1,( S422D)SGK1, SGK2, SGK3 or (T308D,S473D)PKB but not (K127 N)SGK1 significantly stimulated KCNE1-induced current. The effect did not depend on Na(+)/K(+)-ATPase activity. KCNE1-induced current was markedly upregulated by coexpression of KCNQ1 and further increased by additional expression of (S422D)SGK1, SGK2, SGK3 or (T308D,S473D)PKB. In conclusion, all three members of the SGK family of kinases SGK1-3 and protein kinase B stimulate the slowly activating K(+) channel KCNE1/KCNQ1. The kinases may thus participate in the regulation of KCNE1-dependent transport and excitability. PMID- 12634934 TI - Modulation of IP(3)-sensitive Ca(2+) release by 2,3-butanedione monoxime. AB - We describe the actions of 2,3-butanedione monoxime (BDM) on calcium responses in secretory cells. Our studies were prompted by the widespread use of BDM as a myosin-ATPase inhibitor. Application of 10 mM BDM almost completely inhibited agonist-evoked amylase secretion from mouse pancreatic acinar cells. This action might be interpreted as indicating a role for myosin in secretion. However, BDM alone elicited a calcium response in single cells and this calcium signal was sufficient to activate calcium-dependent chloride currents. Furthermore, in some cases, BDM potentiated agonist-evoked calcium signals but almost always blocked agonist-evoked calcium oscillations. These effects of BDM were not due to an action on calcium influx pathways but rather to direct effects on IP(3)-sensitive stores. We conclude that BDM cannot be used for unequivocal identification of the involvement of myosin motors in a cellular response. Further, our evidence suggests that BDM can act directly to modify the opening of IP(3) receptors. PMID- 12634935 TI - Relationships between mechanical power, O(2) consumption, O(2) deficit and high energy phosphates during calf exercise in humans. AB - Whole-body O(2) uptake ( VO(2)), O(2) deficit and the concentration of high energy phosphates (determined by (31)P spectroscopy) in human calf muscle were measured during moderate aerobic square-wave exercise of increasing intensity in ten volunteers. Net VO(2) (above resting) increased linearly with mechanical power, yielding a delta efficiency of 13.1%. "Gross" O(2) deficit increased linearly with net VO(2). The fraction of phosphocreatine (PC) split at steady state increased linearly with the mechanical power and with the O(2) deficit. If the [PC] in resting muscle is known, the slope of the regression between PC split and O(2) deficit (in millimoles) yields the P/O(2) ratio. To calculate this, the O(2) deficit was corrected for the amount of O(2) derived from the body stores, as obtained from literature data. The value so obtained, for a resting [PC] of 30 mM was 5.9, consistent with canonical textbook values. Furthermore, the ratio of "true" O(2) deficit to steady-state VO(2) is a measure of the time constant of VO(2) kinetics at work onset at the muscle level: assuming a monoexponential time course without time delays it amounted to about 17 s, close to the value that can be expected in mammalian muscle at 37 degrees C. PMID- 12634937 TI - Age at disease onset and diagnosis delay in HLA-B27 negative vs. positive patients with ankylosing spondylitis. AB - OBJECTIVE: To investigate differences between HLA-B27(-) and HLA-B27(+) patients with ankylosing spondylitis (AS). METHODS: A total of 1080 patients with AS responded to a questionnaire containing 30 questions; 945 (87.5%) knew their HLA B27 status, 10% of them being B27(-). RESULTS: The average age at disease onset was 27.7 years in B27(-) and 24.8 years in B27(+) AS (P < 0.01). The average age at diagnosis was 39.1 and 33.2 years and the average diagnosis delay 11.4 and 8.5 years, respectively. The distribution in age at disease onset was significantly wider in B27(-) (standard deviation 10.0 years) than in B27(+) AS (8.3 years). The percentages with childhood (age < 16 years) disease onset did not differ significantly (7.6% vs. 6.2%, respectively), whereas the percentage of late onset (age > 40 years) was significantly greater among B27(-) (13%) than among B27(+) (5%) patients with AS. There is a difference in average age at disease onset between male (25.7 years) and female (24.2 years) AS patients, and no difference between patients with primary AS and AS associated with psoriasis, inflammatory bowel disease, or reactive arthritis. Acute anterior uveitis was significantly less frequent in B27(-) (26%) than in B27(+) (41%) patients with AS. CONCLUSIONS. This study of a much larger number of B27(-) AS patients than have been studied previously confirms earlier reports indicating a significantly older average age at disease onset and a less frequent prevalence of acute anterior uveitis in B27( ) than in B27(+) AS. The frequency of late disease onset (after 40 years of age) is significantly higher in B27(-) AS. We provide the first report on significant differences in the distribution curves for the age at disease onset and for the age at diagnosis between B27(-)and B27(+) patients with AS. The average delay between the first spondyloarthritic symptoms and the diagnosis is significantly longer in B27(-) than in B27(+) AS. The frequency of juvenile disease onset (before age 16 years) is nearly the same, irrespective of the B27 status. PMID- 12634938 TI - Predictors of bone mineral density and osteoporosis in patients attending a rheumatology outpatient clinic. AB - The aim of this study was to evaluate determinants of bone mineral density (BMD) and osteoporosis in patients attending our rheumatology outpatient clinic. A cross-sectional analysis of 320 patients with rheumatism (248 noninflammatory and 72 inflammatory arthropathies) was done. Bone mineral density was measured in the lumbar spine (posteroanterior and lateral views) using dual-energy X-ray absorptiometry. Single and multiple analyses were used to assess associations between BMD and potential risk factors including age, height, weight, body mass index (BMI), alkaline phosphatase (ALP), calcium (Ca), phosphorous (P), magnesium (Mg), spot urine calcium/creatinine (Ca/Cr) ratio, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), total cumulative dose and mean daily dosage of prednisolone, and duration of therapy. In these patients, there were significant correlations between BMD and age (r = -0.372, P = 0.000), height (r = 0.308, P = 0.000), weight (r = 0.145, P = 0.011), and ALP (r = -0.262, P = 0.000). By multiple regression analysis, age and ALP were found to be significant predictors of BMD (r(2) = 0.253, P < 0.01). Two hundred eighty-three of the patients had osteoporosis. Multiple logistic regression analysis showed that age and ALP were significant determinants for osteoporosis. These observations suggest that age and ALP were the major determinants of BMD and osteoporosis in these patients, suggesting a contribution of increased bone metabolism to loss of bone in cases of rheumatism. PMID- 12634936 TI - Pathologic thrombopoiesis of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is frequently complicated by thrombocytosis correlated with disease activity. The exact pathogenetic mechanism(s) that cause increased platelet counts in RA are still unknown. Recent investigations indicate that proinflammatory pleiotropic cytokines of RA also have megakaryocytopoietic/thrombopoietic properties. Moreover, several lineage dominant hematopoietic cytokines can also act as acute phase responders and contribute to the inflammation. This review focuses on the current literature and our experience regarding the dual relationships of the pathologic thrombopoiesis of RA. Growth factors contributing to it, namely interleukin (IL)-6, IL-11, stem cell factor, leukemia inhibitory factor, granulocyte colony stimulating factor, thrombopoietin (TPO), and the regulation of megakaryocytopoiesis during the inflammatory cascade are reviewed. Some data indicate that thrombopoietin could contribute to the reactive thrombocytosis of RA. In the non-lineage-specific gp130 cytokine family, IL-6 appears to predominate for the induction of megakaryopoiesis. However, other cytokines and growth factors may also contribute to the pathologic megakaryocytopoiesis of RA. Those pleiotropic mediators seem to act in concert to regulate this enigmatic process. Clarification of the pathobiologic basis of thrombopoiesis in RA may improve understanding of the disease pathogenesis and management of the inflammatory thrombocytosis. PMID- 12634939 TI - Early rheumatoid arthritis patients: relationship of age. AB - The aim of this study was to investigate if age at disease onset comprises a separate parameter for disease expression, prognosis, and outcome in early rheumatoid arthritis (RA) patients. Four hundred thirty-eight patients with early RA (disease duration less than 1 year) were studied. All of them fulfilled the American College of Rheumatology criteria for RA. The demographic, clinical, laboratory, radiologic, and therapeutic characteristics of the disease at diagnosis and during and at the end of follow-up (time period 1981-2000) were analyzed according to age at disease onset (young patients aged less than 60 years at disease onset vs elderly patients aged more than 60 years at disease onset). We found 317 young and 121 elderly patients with early RA. The male:female ratio, which was 1:3.2 in the young patients, was nearly equal in the elderly (1:1.4). In addition, at disease onset elderly patients showed more severe joint involvement (decreased grip strength) associated with high titers of acute phase response (erythrocyte sedimentation rate and C-reactive protein) than the younger patients. However, there were no differences between the two groups in the numbers of tender and swollen joints or acute phase response at the end of the study period. Furthermore, no differences were seen between the two groups concerning the presence of rheumatoid factor. Finally, the two patient groups showed the same degree of radiological changes and functional ability and were treated similarly, except for more frequent corticosteroid use in the elderly. We conclude that elderly patients present with more severe joint involvement at disease onset. However, at the end of the study, no differences were seen concerning radiological changes and functional ability. It seems that age at disease onset does not influence the clinical course and outcome of early RA patients. PMID- 12634940 TI - Expression of the apoptosis accelerator Bax in rheumatoid arthritis synovium. AB - OBJECTIVE: The aim of this study was to analyze the expression of apoptosis related molecules in rheumatoid arthritis (RA) synovium, with special emphasis on the apoptosis accelerator Bax. METHODS: Immunohistochemical analysis of Bax, Bcl 2, and Bcl-x(L) was performed in tissue specimens of patients with RA and compared to normal synovial tissue. Expression of Bax was additionally determined by double labeling with CD68, p53, and Ki-67 (clone MIB-1). Apoptotic cells were further identified by the terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) method. RESULTS: In RA, expression of Bax was higher than in healthy controls and occurred in CD68-positive and -negative synoviocytes. Strong Bax staining was also found in chondrocytes at sites of cartilage degradation. Bax-positive synoviocytes could be detected with p53 and also with Ki-67. Bax and Bcl-x(L) were markedly colocalized in synovium. The TUNEL method revealed only few positive synoviocytes. CONCLUSIONS: The marked colocalization of Bax and antiapoptotic Bcl-x(L) as well as the low frequency of TUNEL-positive cells in RA synovium suggest that Bax activity is not sufficient to decrease synovial hyperplasia in RA. Apoptotic mechanisms in RA chondrocytes might also be important for the pathogenesis of joint damage. PMID- 12634941 TI - The efficacy of lumbar traction in the management of patients with low back pain. AB - The literature on the efficacy of traction in the treatment of low back pain (LBP) is conflicting. The aim of this study was to examine its efficacy in this disorder. Forty-two patients with at least 6 weeks of nonspecific LBP were selected. Demographic data were obtained. All patients completed the Oswestry disability index (ODI) to assess disability and the 10-cm visual analog scale (VAS) for evaluation of pain. Subjects were randomly assigned into group 1, receiving only standard physical therapy, or group 2, receiving standard physical therapy with conventional lumbar traction. Standard physical therapy consisted of local heat, ultrasound for the lumbar region, and an active exercise program, given for ten sessions in all. The subjects received instruction on correct posture and recommended therapeutic exercises. They were reevaluated at the end of treatment and at 3-month follow-up. The mean outcome measures were global improvement and satisfaction with the therapy, as well as disability by ODI and pain by VAS. There were no group differences in terms of demographic and baseline clinical characteristics. There was a significant reduction in pain intensity and disability at the end of treatment in both groups. There was complete or mild improvement in 47.6% of group 1 and 40% of group 2. The satisfaction rate with both treatments was more than 70% immediately after the therapies. During the 3 month period, the outcome measures except disability remained statistically stable, with no difference amongst groups. Disability was significantly reduced at follow-up in both groups. Of the patients, 51% continued with the recommended exercises and had significantly lower disability scores than those who did not continue with the exercises. Pain and global improvement were also better in this group, but the difference was not statistically significant. In conclusion, no specific effect of traction on standard physical therapy was observed in our study group. We suggest focusing on back education and exercise therapy in the management of patients suffering from this chronic condition. PMID- 12634942 TI - Body composition, bone mineral density, and circulating leptin levels in postmenopausal Turkish women. AB - OBJECTIVE: We analyzed the relationship between serum leptin levels and bone mineral density (BMD) values as well as the relationship between serum leptin levels and whole body composition, whether or not they were associated. In addition, we also investigated whether lean mass or fat mass is a better predictor of BMD in postmenopausal Turkish women. DESIGN AND MEASUREMENTS: One hundred consecutive postmenopausal women with a mean age of 55.1 +/- 6.3 years who visited our outpatient clinic for the evaluation of osteoporosis were recruited. Skin fold thickness at four sites and waist:hip ratio were measured. Body mass index (BMI) was calculated in kg/m(2). Serum concentrations of leptin, insulin, and estradiol were evaluated. Bone formation and resorption markers were also determined. The BMD values were measured by dual energy X-ray absorptiometry (DEXA) at the lumbar spine and femoral neck. Whole body composition (lean mass, fat mass, and percentage of fat), total bone mineral content (BMC) in g, and total BMD were also measured by DEXA. RESULTS: Serum leptin levels did not correlate with BMD values at all skeleton sites measured. Leptin correlated positively with fat mass, percentage of fat, and BMI (r = 0.738 and P = 0.00, r = 0.536 and P = 0.00, r = 0.356 and P = 0.00, respectively). Lean mass correlated with BMD at all sites measured (r = 0.339 and P = 0.00, r = 0.312 and P = 0.01, r = 0.523 and P = 0.00, r = 0.636 and P = 0.00). Lean mass correlated with BMI (r = 0.636, P = 0.00) but not with serum leptin (r = -0.021, P = 0.881), and it was an independent determinant of BMD at all skeleton sites measured. CONCLUSION: Our study showed that lean mass is a better predictor than fat mass of bone mineral density and that serum leptin levels are not associated with BMD. PMID- 12634943 TI - Life-threatening extrarenal lupus in children despite improvement in serologic findings. AB - Systemic lupus erythematosus (SLE) is an autoimmune disorder with the potential for multiorgan involvement. Serologic tests are helpful in establishing the diagnosis of SLE and predicting disease flares. However, there are few data on the relationship between the onset of new organ involvement and lupus serologies, especially in children. This report details our experience in managing two children with lupus nephritis. Both developed life-threatening extrarenal complications (cerebritis and carditis) soon after receiving high-dose immunosuppressive therapy and despite normalizing serologies. This lack of concordance between serologies and the development of carditis and cerebritis needs to be recognized so that health care professionals treating children with SLE can promptly intensify immunosuppressive medications and avoid life threatening delays from seeking alternative explanations for symptomatology. PMID- 12634944 TI - Severe livedo vasculitis treated with potassium iodide. AB - A young woman presenting severe livedo vasculitis (LV) was successfully treated with potassium iodide (KI). Previous attempts to cure her disorder with corticosteroids, low-dose aspirin, and azathioprine in various combinations had failed. To our knowledge, this is the first report of LV treated with KI. PMID- 12634945 TI - Monitoring minimal residual disease in AML: the right time for real time. AB - Detection of minimal residual disease (MRD) by polymerase chain reaction (PCR) has become an essential tool for molecular monitoring of acute myeloid leukemia (AML). Currently, specific translocation markers are available for 40-50% of AMLs. Expression markers may widen this spectrum to 70-90%. Quantitative PCR (Q PCR, real-time PCR) is now as sensitive as conventional two-step PCR and could improve as well as facilitate clinical decision-making. Q-PCR has been applied to a variety of molecular markers, delineating threshold levels early after induction therapy, for postinduction monitoring, as well as for the detection of relapse. For most markers, lack of decline of transcript levels by less than 2 logs after chemotherapy has been established as a poor prognostic sign. Moreover, increases in transcript levels are almost invariably associated with relapse. However, the predictive value of PCR negativity after chemotherapy is not as clear. The major tasks for the future will be standardization of Q-PCR techniques, exact definition of threshold levels, and monitoring schedules in bone marrow (BM) and peripheral blood (PB), as well as investigation of novel markers found by microarray analysis. PMID- 12634946 TI - Chronic myeloproliferative disorders with thrombocythemia: a comparative study of two classification systems (PVSG, WHO) on 839 patients. AB - A multicenter observational study was performed on 839 adult patients with a chronic myeloproliferative disorder and a platelet count in excess of 600 x 10(9)/l to compare the updated criteria of the Polycythemia Vera Study Group (PVSG) with the recently published WHO classification. Essential thrombocythemia (ET) was diagnosed in 483 patients according to the PVSG; however, when considering histopathology as a major diagnostic feature of the WHO criteria, (true) ET could be established in only 162 patients. The remaining cases were found to represent either initially prefibrotic (184 patients) or early fibrotic (137 patients) chronic idiopathic myelofibrosis (IMF). On the other hand, both classification systems enabled a clear-cut distinction of patients showing overt IMF and polycythemia vera. Follow-up examinations in 140 patients with ET according to the PVSG criteria included also sequential bone marrow biopsies (interval: 38+/-30 months). A transition into mild reticulin fibrosis occurred in only 2 of 49 patients with (true) ET in contrast to 45 of 91 patients with initial and early IMF where a progression into overt myelofibrosis was encountered. Survival patterns for ET displayed significant differences because according to the PVSG a 16.5% disease-specific loss of life expectancy was calculable compared to a value of only 8.9% when following the WHO criteria. Contrasting this finding, initial and early IMF mimicking ET was characteriZed by a reduction of life expectancy ranging between 21.6% and 32.3 %. In conclusion, a more accurate classification of ET is warranted by regarding the WHO criteria that include histopathology as a major feature for diagnosis. PMID- 12634947 TI - Aspirin-responsive painful red, blue, black toe, or finger syndrome in polycythemia vera associated with thrombocythemia. AB - Five patients with red, purple blue, or black toes or fingers due to thrombocythemia associated with polycythemia vera (polycythemia and thrombocythemia vera) in four and essential thrombocythemia (thrombocythemia vera) in one are described. The microvascular erythromelalgic syndrome of thrombocythemia was overlooked and progressed to cold blue swollen and painful fingers or black toes in three patients with polycythemia and thrombocythemia vera due to arteriographically documented occlusions of digital or large peripheral arteries with no evidence of preexistent atherosclerotic vascular disease. Concomitant erythromelalgia of the hand palm could be confirmed by the histopathological findings of arteriolar thrombotic lesions in the reticular dermis in two patients with polycythemia and thrombocythemia vera. The increased hematocrit in the presented patients with polycythemia and thrombocythemia vera contributed to the progression of the microvascular syndrome of thrombocythemia to major occlusive ischemic events of the extremities. Standard therapy with oral anticoagulants and reduction of the hematocrit to normal by bloodletting did not affect the platelet-mediated microvascular erythromelalgic, ischemic symptoms in the patients with polycythemia vera because thrombocythemia vera persisted. Complete relief of pain and restoration of the ischemic acral circulation disturbances in patients with thrombocythemia vera or thrombocythemia associated with polycythemia vera in maintained remission by bloodletting could be obtained by long-term treatment with low-dose aspirin. PMID- 12634948 TI - Incidence of tumor lysis syndrome in children with advanced stage Burkitt's lymphoma/leukemia before and after introduction of prophylactic use of urate oxidase. AB - To evaluate the clinical benefit of the prophylactic use of urate oxidase in children with non-Hodgkin's lymphoma (NHL), we analyzed the incidence and complications of tumor lysis syndrome (TLS) in children with B-cell acute lymphoblastic leukemia (B-ALL) or stage III/IV Burkitt's lymphoma and a lactate dehydrogenase (LDH) level > or =500 U/l before and after the introduction of a protocol amendment to use urate oxidase for the prophylaxis of TLS. Data from 1791 children with NHL enrolled in the two subsequent multicenter studies NHL-BFM 90 and 95 were evaluated. The presence of the side effects TLS, anuria, sepsis, and other complications during the first 2 weeks after admission were registered. Until March 1996, no urate oxidase was used (period 1). From November 1997 all children with B-ALL or stage III and IV B-NHL and LDH > or =500 U/l should receive urate oxidase prophylactically (period 3). In between (period 2), urate oxidase was given in a minority of hospitals therapeutically. Initial chemotherapy was identical. Altogether, 78 children (4.4%) developed a TLS. Patients with B-ALL had the highest risk to develop a TLS (26.4%) followed by B ALL/Burkitt's lymphoma and a LDH > or =500 U/l (14.9%). In period 1, 16.1% and 9.2% of the latter children developed a TLS or anuria, respectively, compared to 12.3% and 6.2% in period 3 ( p=NS). The incidence of sepsis remained unchanged (5.0% vs 4.6%). In children with B-ALL the differences in the incidence of TLS and anuria between period 3 and period 1 were more pronounced, reaching significance for anuria (15.4% vs 3.8%, p=0.03). Our results suggest that patients with the highest risk to develop a TLS might benefit from the prophylactic use of urate oxidase. PMID- 12634949 TI - Increased frequency of CD3/8/56-positive umbilical cord blood T lymphocytes after allo-priming in vitro. AB - Umbilical cord blood (UCB) or adult peripheral blood mononuclear cells (PBMC) were repeatedly stimulated by HLA-mismatched allogeneic Epstein-Barr virus (EBV) transformed cell lines, and the bulk responders or single-cloned cells were immunophenotypically analyzed by flow cytometry. One month after the allo stimulation, not in PBMC but in UCB, the proportion of CD3/8/56 triple positive T lymphocytes significantly increased. Furthermore, UCB clones exhibited those unique CD3/8/56 markers at an extremely higher frequency than PB clones. They showed as much strong killing activity against allo-stimulators as conventional PB CD56-negative, CD8 T-cell clones, whereas they did not kill the other target, Raji cells. UCB CD3/8/56 T cell clones produced a smaller amount of interferon gamma compared with PB CD4 or PB CD8 T cell clones. We concluded that CD8 T cells coexpressed with CD56 marker expanded after allo-priming in vitro and would become one of the graft-versus-host (GVH) effectors after UCB transplantation. PMID- 12634950 TI - Erythropoiesis and serum sVCAM-1 levels in adults with sickle cell disease. AB - Sickle cell patients are characterized by stress erythropoiesis involving cytokines, growth factors, and adhesion molecules. We set out to determine whether serum soluble vascular cell adhesion molecule-1 (sVCAM-1) levels, which are inversely related to red blood cell counts in sickle cell disease (SCD), reflect erythropoietic activity in adult HbSS patients. Serum levels of sVCAM-1 were compared to erythropoietin (EPO), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), and soluble transferrin receptor (sTfR) levels in 29 adults with HbSS, and their respective levels were also compared to 29 race- and age-matched HbAA controls. EPO and sTfR levels were increased as compared to healthy controls, whereas IL-3 and GM-CSF were not. No significant correlation of sVCAM-1 levels could be detected with any of the measured erythropoietic markers. Patients, but not controls, with detectable IL-3 levels had lower sTfR and GM-CSF levels as compared to patients with undetectable IL-3 levels. Even though a link of sVCAM-1 to erythropoiesis could not be established, it cannot be ruled out that sVCAM-1 levels reflect the release of young red blood cells into the circulation. IL-3 and GM-CSF levels suggest that different rates of erythropoiesis may be characterized by specific cytokine profiles in SCD. Further research should focus on the potential cytokines and adhesion molecules involved in sickle cell erythropoiesis, as this may increase our understanding of sickle cell complications and may provide us with potential markers for risk assessment in sickle cell disease as well. PMID- 12634951 TI - A novel splicing acceptor mutation of the factor VIII gene producing skipping of exon 25. AB - A gross deletion in the factor VIII (FVIII) mRNA was determined by reverse transcriptase polymerase chain reaction (RT-PCR) for a patient with moderately severe hemophilia A. Sequencing of the RT-PCR product depicted a 177-bp deletion ranging from nucleotide (nt) 6724 to nt 6900 of FVIII cDNA, exactly corresponding to the whole exon 25. Further study of the genomic DNA revealed the presence of a single base pair substitution (G >A) at position -1 of intron 24. The absolute consensus AG doublet of the intron 24 splicing acceptor changed to AA. In the novel splice site mutation, exon 24 was erroneously spliced to exon 26, skipping exon 25. The FVIII antigen level was normal despite the markedly reduced functional activity. Since exon 25 corresponds to part of the C2 domain, we speculate that for this patient the aberrant C2 domain markedly reduces binding affinity of FVIII protein to the phospholipid membrane, thus severely impairing the protein function. PMID- 12634952 TI - Familial hyperhomocysteinemia: multiple venous thrombosis in four generations of a family. AB - We describe a family in which four generations (eight members) had deep vein thrombosis of the lower limb and three of the alive members had documented hyperhomocysteinemia. In addition, one of the family members had evidence of arterial thrombosis in the form of cerebral infarcts. Interestingly, all affected members in the family were males. PMID- 12634953 TI - Two new mutations in cis on (G)gamma chain of fetal hemoglobin: Hb F-Madrid [G gamma 50(D1)Ser-->Cys] and [G gamma 75(E19)Ile-->Thr]. AB - We describe a new structural hemoglobin variant of (G)gamma with two amino acid replacements in cis found in the umbilical cord blood of a neonate in Madrid, Spain. The substitutions were identified on exon 2 of the (G)gamma globin gene, at codon 50 (T CT-->T GT) and at codon 75 (A TA-->A CA). We have named it Hb F Madrid. The father of the propositus was the carrier of the same (G)gamma chain variant and, moreover, molecular study of alpha genes revealed the loss of an alpha gene (-alpha(3.7)/alpha alpha) both in the propositus and his mother. PMID- 12634954 TI - Zidovudine-related anemia with reticulocytosis. AB - In patients infected with human immunodeficiency virus (HIV), zidovudine has been known to cause a severe hypoproliferative anemia that resolves promptly when the drug is stopped. Some patients, however, may come to medical attention after having stopped taking the drug for 1-2 weeks and may already be in the recovery phase and present with severe anemia, normoblastemia, and reticulocytosis. These features can be mistaken for hemolytic disease, as shown by our experience with three such patients. PMID- 12634955 TI - Fatal Kikuchi-Fujimoto disease: the lupus connection. AB - Kikuchi-Fujimoto disease (KFD) is a histiocytic necrotizing lymphadenitis found mainly in young women. Patients typically present with cervical lymphadenopathy and fever and follow almost always a benign course with excellent outcomes. The etiology of KFD remains unknown and controversial although several viruses have been associated with this disease. One theory proposes that KFD may be a self limiting form of systemic lupus erythematosus (SLE). This theory is strongly supported by the fact that microscopic features of KFD can be very similar to those found in lupus lymphadenitis. Despite its usually benign course, several cases with fatal outcomes have been reported. We report here a case of clinically aggressive KFD, which featured several autoimmune-related events and resulted in a fatal outcome. Autopsy studies showed characteristic findings of SLE, which suggests a remarkable link between SLE and KFD. Early and intensive immunosuppressive treatment may be the only option for patients who develop very aggressive forms of KFD in order to avoid a fatal outcome. PMID- 12634956 TI - Allogeneic stem cell transplant from HLA-identical sibling for chronic granulomatous disease and review of the literature. AB - Chronic granulomatous disease (CGD) is a rare primary immunodeficiency caused by an abnormal function of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in the phagocytic cells, which results in an increased susceptibility to severe bacterial and fungal infections. We report on a 12-year-old boy with X linked CGD who was successfully treated with allogeneic bone marrow transplantation from an HLA-identical sibling following a conditioning regimen consisting of busulphan (BU, 16 mg/kg) and cyclophosphamide (CY, 200 mg/kg). At >2 years from transplant, the boy is in excellent clinical and hematological condition with full chimerism. Our patient is the 24th case of CGD transplanted from an HLA-identical sibling. A review of the literature revealed that 20 of 24 CGD patients are alive and disease free 1-7 years after transplant. Most of these patients were conditioned with the BUCY combination, which should be considered the recommended regimen. PMID- 12634957 TI - Successful reintroduction of methotrexate after acute pneumonitis in a patient with acute lymphoblastic leukemia. AB - Low-dose methotrexate (MTX) is used as disease-modifying therapy in severe rheumatoid arthritis and as maintenance treatment in patients with complete remission of acute lymphoblastic leukemia (ALL). It is generally well tolerated, but in 27% of patients acute pneumonitis leads to discontinuation of treatment. We describe a 56-year-old female patient with newly diagnosed pre-B-ALL. She was treated with induction chemotherapy in July 1999 which lead to complete remission. Maintenance treatment with low-dose MTX and 6-mercaptopurine (6-MP) was started in December 1999. In April 2000 she was hospitalized because of fever, cough, and rapidly progressive dyspnea. No pathogens could be cultured from blood or bronchoalveolar lavage fluid. Computed tomography of the lungs revealed interstitial infiltration and ground-glass opacities. Acute pneumonitis was diagnosed, and MTX was stopped. Prednisone therapy lead to rapid clinical amelioration of dyspnea and hypoxemia. Since for this patient there was no alternative leukemia therapy, MTX was successfully reintroduced in August 2000 without reappearance of any respiratory symptoms. We discuss risk profile, clinical and histological presentation, and therapy of MTX-induced pneumonitis. To our knowledge, this is the first patient with ALL in whom successful reintroduction of MTX after severe pneumonitis has been reported. PMID- 12634958 TI - Primary gastric T-cell lymphoma not associated with human T-lymphotropic virus type I: a case report and review of the literature. AB - Primary gastric T-cell lymphoma (PGTL) not associated with human T-lymphotropic virus type I (HTLV-I) is extremely rare and such a case is reported herein. The patient was a 58-year-old Japanese male presenting with submucosal tumor of the stomach identified on endoscopic examination. The lesion was diagnosed as non Hodgkin's lymphoma by endoscopic biopsy and classified as peripheral T-cell lymphoma, unspecified, due to clonal rearrangement of the T-cell receptor beta (TCR) gene and expression of TCR beta protein in the absence of B-cell genotypes and phenotypes. Unlike previously reported cases of HTLV-I-unassociated PGTL, lymphoma in the current case was characterized histologically as "low grade" and phenotypically as CD4+, TIA-1+, granzyme B+, and CD103-. The lymphoma responded well to chemotherapy and radiation, and the patient was well with no detectable disease 10 months after initiation of therapy. A review of patients with PGTL in the literature revealed a few long-term survivors, and the investigation of therapeutic strategies for PGTL is, therefore, necessary. PMID- 12634960 TI - A hand-held imaging probe for radio-guided surgery: physical performance and preliminary clinical experience. AB - Improvements in the specificity of radiopharmaceutical compounds have been paralleled by an upsurge of interest in developing small detectors to assist surgeons in localizing tumour tissue during surgery. This study reports the main technical features and physical characteristics of a new hand-held gamma camera dedicated to accurate and real-time intra-operative imaging. First clinical experience is also reported. The POCI (Per-operative Compact Imager) camera consists of a head module composed of a high-resolution interchangeable lead collimator and a CsI(Na) crystal plate optically coupled to an intensified position-sensitive diode. The current prototype has a 40-mm diameter field of view, an outer diameter of 9.5 cm, a length of 9 cm and a weight of 1.2 kg. Overall detector imaging characteristics were evaluated by technetium-99m phantom measurements. Three patients with breast cancer previously scheduled to undergo sentinel lymph node detection were selected for the preliminary clinical experience. Preoperative images of the lymphatic basin obtained using the POCI camera were compared with conventional transcutaneous explorations using a non imaging gamma probe. The full-width at half-maximum (FWHM) spatial resolution was investigated in both air and scattering medium; when the phantom was placed in contact with the collimator, the POCI camera exhibited a 3.2 mm FWHM. The corresponding sensitivity was 290 cps/MBq. The preliminary clinical results showed that POCI was able to predict the number and location of all SLNs. In one case, two deep radioactive nodes missed by the gamma probe were detected on the intra-operative images. This very initial experience demonstrates that the physical performance of the POCI camera is adequate for radio-guided surgery. These results are sufficiently encouraging to prompt further evaluation studies designed to determine the specific and optimal clinical role of intra-operative imaging devices. PMID- 12634961 TI - CT-based attenuation correction in the calculation of semi-quantitative indices of [18F]FDG uptake in PET. AB - The introduction of combined PET/CT systems has a number of advantages, including the utilisation of CT images for PET attenuation correction (AC). The potential advantage compared with existing methodology is less noisy transmission maps within shorter times of acquisition. The objective of our investigation was to assess the accuracy of CT attenuation correction (CTAC) and to study resulting bias and signal to noise ratio (SNR) in image-derived semi-quantitative uptake indices. A combined PET/CT system (GE Discovery LS) was used. Different size phantoms containing variable density components were used to assess the inherent accuracy of a bilinear transformation in the conversion of CT images to 511 keV attenuation maps. This was followed by a phantom study simulating tumour imaging conditions, with a tumour to background ratio of 5:1. An additional variable was the inclusion of contrast agent at different concentration levels. A CT scan was carried out followed by 5 min emission with 1-h and 3-min transmission frames. Clinical data were acquired in 50 patients, who had a CT scan under normal breathing conditions (CTAC(nb)) or under breath-hold with inspiration (CTAC(insp)) or expiration (CTAC(exp)), followed by a PET scan of 5 and 3 min per bed position for the emission and transmission scans respectively. Phantom and patient studies were reconstructed using segmented AC (SAC) and CTAC. In addition, measured AC (MAC) was performed for the phantom study using the 1-h transmission frame. Comparing the attenuation coefficients obtained using the CT- and the rod source-based attenuation maps, differences of 3% and <6% were recorded before and after segmentation of the measured transmission maps. Differences of up to 6% and 8% were found in the average count density (SUV(avg)) between the phantom images reconstructed with MAC and those reconstructed with CTAC and SAC respectively. In the case of CTAC, the difference increased up to 27% with the presence of contrast agent. The presence of metallic implants led to underestimation in the surrounding SUV(avg) and increasing non-uniformity in the proximity of the implant. The patient study revealed no statistically significant differences in the SUV(avg) between either CTAC(nb) or CTAC(exp) and SAC reconstructed images. The larger differences were recorded in the lung. Both the phantom and the patient studies revealed an average increase of approximately 25% in the SNR for the CTAC-reconstructed emission images compared with the SAC reconstructed images. In conclusion, CTAC(nb) or CTAC(exp) is a viable alternative to SAC for whole-body studies. With CTAC, careful consideration should be given to interpretation of images and use of SUVs in the presence of oral contrast and in the proximity of metallic implants. PMID- 12634962 TI - Quantitation in PET using isotopes emitting prompt single gammas: application to yttrium-86. AB - Several yttrium-90 labelled somatostatin analogues are now available for cancer radiotherapy. After injection, a large amount of the compound is excreted via the urinary tract, while a variable part is trapped in the tumour(s), allowing the curative effect. Unfortunately, the compound may also be trapped in critical tissues such as kidney or bone marrow. As a consequence, a method for assessment of individual biodistribution and pharmacokinetics is required to predict the maximum dose that can be safely injected into patients. However, (90)Y, a pure beta(-)particle emitter, cannot be used for quantitative imaging. Yttrium-86 is a positron emitter that allows imaging of tissue uptake using a PET camera. In addition to the positron, (86)Y also emits a multitude of prompt single gamma rays, leading to significant overestimation of uptake when using classical reconstruction methods. We propose a patient-dependent correction method based on sinogram tail fitting using an (86)Y point spread function library. When applied to abdominal phantom acquisition data, the proposed correction method significantly improved the accuracy of the quantification: the initial overestimation of background activity by 117% was reduced to 9%, while the initial error in respect of kidney uptake by 84% was reduced to 5%. In patient studies, the mean discrepancy between PET total body activity and the activity expected from urinary collections was reduced from 92% to 7%, showing the benefit of the proposed correction method. PMID- 12634963 TI - Identification of the sentinel lymph node in patients with malignant melanoma: what are the reasons for mistakes? AB - Scintigraphic identification of the sentinel lymph node is achievable in nearly all patients with malignant melanoma. However, in a very small number of cases the sentinel node fails to be detected, and sometimes recurrence appears during follow-up in patients who had previously tested negative for metastatic disease. The purpose of this study was to review our experience in order to isolate the reasons for erroneous sentinel lymph node identification. The evaluation involved 435 consecutive malignant melanoma patients with AJCC stages I and II (clinically negative nodes) and Breslow thickness >0.76 mm. Lymphoscintigraphy was performed the day before surgery by intradermal administration of technetium-99m labelled nanocolloid. Dynamic and static images were obtained. The sentinel node was intraoperatively identified with the aid of patent blue dye and a hand-held gamma probe. After removal, routine histopathological examination with haematoxylin eosin (H-E) and immunohistochemistry with S 100 and HMB45 (IHC) were performed. In those patients who developed regional recurrences during follow-up, sentinel nodes were further evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR). Lymphoscintigraphy visualised at least one sentinel node in 434 out of 435 patients (99.8%). Uptake in in-transit sentinel lymph nodes was observed in 32 patients (7.4%). During surgery, localisation and removal of sentinel nodes was successful in 430/435 patients (98.8%). A total of 790 sentinel lymph nodes were harvested, with a mean of 1.8 per patient. Routine histopathological examination with H-E or IHC revealed metastatic disease in 72 patients (16.8%). During a mean follow-up of 26 months, seven of those patients with a negative sentinel node developed regional lymph node metastases. In five of them RT-PCR was positive for micrometastases within the sentinel node. In conclusion, erroneous sentinel lymph node identification can be due to changes in the surgical team, difficult lymph node location or absence of a thorough histological study. Nevertheless, it is not possible to explain completely why, in a very small percentage of cases, the sentinel node is erroneously identified. PMID- 12634964 TI - Tumour dosimetry and response in patients with metastatic differentiated thyroid cancer using recombinant human thyrotropin before radioiodine therapy. AB - The development of recombinant human thyrotropin (rhTSH) has given clinicians new options for diagnostic follow-up and treatment of patients with differentiated thyroid cancer (DTC). This paper evaluates the tumour dosimetry and response following -iodine-131 treatment of metastatic thyroid cancer patients after rhTSH stimulation instead of classical hormone withdrawal-induced hypothyroidism. Nineteen consecutive (131)I treatments in 16 patients were performed after rhTSH stimulation. All patients had undergone a near-total thyroidectomy followed by an ablative dosage of (131)I. They all suffered from metastatic or recurrent disease showing tumoral (131)I uptake on previous post-treatment scintigraphy. Dosimetric calculations were performed using (131)I tumour uptake measurements from post treatment (131)I scintigrams and tumour volume estimations from radiological images. Response was assessed by comparing pre-treatment serum thyroglobulin (Tg) level with the Tg level 3 months post treatment. In 18 out of 19 treatments, uptake of (131)I in metastatic or recurrent lesions was seen. The median tumour radiation dose was 26.3 Gy (range 1.3-368 Gy), and the median effective half-life was 2.7 days (range 0.5-6.5 days). Eleven of 19 treatments (10/16 patients) were evaluable for response after 3 months. (131)I therapy with rhTSH resulted in a biochemical partial response in 3/11 or 27% of treatments (two patients), biochemical stable disease in 2/11 or 18% of treatments and biochemical progressive disease in 6/11 or 55% of treatments. Our study showed that although tumour doses in DTC patients treated with (131)I after rhTSH were highly variable, 45% of treatments led to disease stabilisation or partial remission when using rhTSH in conjunction with (131)I therapy, without serious side-effects and with minimal impact on quality of life. RhTSH is therefore adequately satisfactory as an adjuvant tool in therapeutic settings and is especially suitable in advanced recurrent or metastatic DTC patients who may be intolerant to TSH stimulation by levothyroxine withdrawal. PMID- 12634965 TI - 99mTc-interleukin-2 and (99m)Tc-HMPAO granulocyte scintigraphy in patients with inactive Crohn's disease. AB - Crohn's disease (CD) is a chronic inflammatory bowel disease that may involve the whole gut. Marked intestinal T cell and macrophage activation is a key feature of the disease. Polymorphonuclear cell infiltration is also observed in the diseased gut, mainly during active inflammation. Scintigraphic detection of granulocytes and activated lymphocytes infiltrating the gut wall may be useful in identifying a subgroup of patients with clinically inactive CD who are undergoing early clinical relapse. The aims of the present study were (a) to compare the effectiveness of scintigraphy with (99m)Tc-labelled interleukin-2 ((99m)Tc-IL2) and with (99m)Tc-HMPAO labelled granulocytes ((99m)Tc-WBC) in detecting the presence and extent of bowel inflammation in patients with long-term inactive CD (>12 months) and (b) to assess the accuracy of these techniques in predicting future disease relapse. We studied 29 patients with ileal and/or colonic CD in stable clinical remission (Crohn's Disease Activity Index <150 for at least 12 months) using both (99m)Tc-IL2 and (99m)Tc-WBC scintigraphy in order to evaluate the extent of acute and chronic inflammation in the bowel. Planar and single photon emission tomography images were acquired in each patient at 1 h p.i. For quantitative analysis of (99m)Tc-IL2 uptake, the abdomen was divided into 32 regions of interest. Despite the absence of symptoms, 18 patients (62%) showed a positive (99m)Tc-IL2 and 18 (62%) a positive (99m)Tc-WBC scan. Only 12 patients (41.4% of the total group) were positive on both scans, and the sites of IL2 and granulocyte bowel uptake were usually located in different segments, indicating that in CD, acute and chronic inflammation can be present in different sites. As far as the prognostic role of the two scans in predicting future disease relapse is concerned, both (99m)Tc-IL2 and (99m)Tc-WBC scintigraphy showed a high negative predictive value (1.00 and 0.91, respectively) but a weak positive predictive value (0.44 and 0.39, respectively). Nevertheless, Kaplan-Meier curves generated between scintigraphic findings and time free from disease relapse were statistically different only for (99m)Tc-IL2 scintigraphy (log-rank test, P=0.013). These results indicate that (99m)Tc-IL2 scintigraphy can be useful in selecting CD patients in clinical remission who could benefit from preventive therapy to avoid disease relapse. PMID- 12634966 TI - Quantitative estimation of myocardial salvage after primary percutaneous transluminal coronary angioplasty in patients with angiographic no reflow. AB - Angiographic Thrombolysis in Myocardial Infarction (TIMI) flow grade 0.75) while the correlations between PM and Cu/Zn/V were modest (R = 0.2-0.6). These results from controlled human exposure linked specific PM components to pulmonary neutrolphil influx and blood fibrinogen increase, and indicated the soluble components of pollutant particles may differentially affect pulmonary and hematological systems in humans exposed to PM. PMID- 12635003 TI - Effects of inhaled CdO particles on the sphingolipid synthesis of rat lungs. AB - Surfactant lipids of the alveolar space protect the lung from various environmental stimuli. We investigated the influence of ultrafine (UF) CdO particles inhalation on two key enzymes involved in lung sphingolipid metabolism, serine palmitoyltransferase (SPT), and sphingomyelinase (SMase). Rats inhaled either 0.63 mg UF-CdO/m(3) for 6 h (group 1), or 1.08 mg UF-CdO/m(3) 12 h/day for 10 days (group 2). Two corresponding control groups inhaled filtered clean air. Additional rats intratracheally instilled with lipopolysaccharide (LPS) were used as positive controls. Semiquantitative reverse-transcription polymerase chain reaction (RT-PCR) of lung tissue showed a significant increase in the level of SPT mRNA (LCB2 subunit) expression in group 2 compared to the corresponding controls (p <.01). Group 1 and LPS were not statistically different from control. No alteration in the mRNA level of SMase was detected in any exposure group. The immunohistochemical analysis showed that SPT (LCB2 subunit) localization was stronger in the alveolar type II cells of group 2 lungs compared to the corresponding controls. These results were correlated with alterations in BALF cellular and biochemical parameters and lung morphology. Since SPT is the key enzyme for de novo sphingolipid synthesis in lung surfactant and SMase is responsible for sphingomyelin catabolism, we can postulate that high-dose UF-CdO exposure for 10 days induces an increase in sphingolipid synthesis in the type II cells of rat lungs that would not be promptly followed by its degradation. PMID- 12635004 TI - Short-term inhalation of particulate transition metals has little effect on the electrocardiograms of dogs having preexisting cardiac abnormalities. AB - There is growing epidemiological evidence for statistical associations between increases in air pollution, especially particulate matter, and increases in cardiovascular morbidity and mortality. Laboratory studies have shown that transition metals contribute strongly to the effects of high lung doses of model particles on changes in the electrocardiograms of animals. The present study evaluated the effects of short-term inhalation exposure to respirable particles of specific oxide and sulfate forms of transition metals on heart rate and the electrocardiogram of old dogs having preexisting cardiac abnormalities. Conscious beagle dogs were exposed by oral inhalation for 3 h on each of 3 successive days to aerosols of manganese, nickel, vanadium, iron, and copper oxides, and nickel and vanadium sulfates as single compounds at concentrations of 0.05 mg/m(3). Electrocardiograms were recorded and evaluated for exposure-related changes in heart rate, heart rate variability, and abnormalities of waveforms. Although the electrocardiograms of this population of dogs having potential age and cardiovascular susceptibility factors reflected their underlying clinical abnormalities, no significant effect of exposure to the transition metal aerosols was observed. PMID- 12635006 TI - Oxidative damage of sulfur dioxide inhalation on stomachs and intestines of mice. AB - Effects of exposure to sulfur dioxide (SO(2)) on levels of thiobarbituric acid reactive substances (TBARS), levels of reduced glutathione (GSH), and the activities of Cu,Zn-superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) were investigated in stomachs and intestines of Kunming albino mice of both sexes. SO(2) exposure at different concentrations (22, 56, and 112 mg/m(3)) was administered to the animals of SO(2) groups in the exposure chambers for 6 h/day for 7 days, while control groups were exposed to filtered air under the same condition. Our results show that SO(2) caused lipid peroxidation and changes of antioxidative status in stomachs and intestines of mice. Exposure to SO(2) at all concentrations tested caused significantly the increase of TBARS levels in stomachs and intestines of mice. For the stomachs, activities of these antioxidant enzymes and levels of GSH were significantly unaltered by SO(2) at low concentrations, except significant increase of SOD activity in the stomachs of male mice. However, higher SO(2) caused the significant increases of CAT activities and the significant decreases of GSH levels and activities of SOD and GPx. For intestines, SO(2) at all concentrations tested decreased significantly activities of SOD relative to control animals; SO(2) caused the decreases of GPx activities, but only the decreases of GPx activities caused by SO(2) exposures at higher concentrations (56 and 112 mg/m(3)) were statistically significant. SO(2) at all concentrations tested tended to increase the CAT activities in a dose dependent manner, but the decreases of CAT activities caused by higher SO(2) were significant. SO(2) at all concentrations tested decreased significantly levels of GSH in intestines of both sexual mice. These results lead to conclusion that SO(2) exposure can caused oxidative damage to stomachs and intestines of mice, and SO(2) is a toxic agent to stomachs and intestines of mammals, not only to respiratory system. Further work is required to understand toxicological role of SO(2) on multiple or even all organs in human and animal. PMID- 12635005 TI - Evaluation of relationships between mainstream smoke acetaldehyde and "tar" and carbon monoxide yields in tobacco smoke and reducing sugars in tobacco blends of U.S. commercial cigarettes. AB - Mainstream (MS) smoke acetaldehyde (AA), carbon monoxide (CO), and "tar" yields and percentage total reducing sugars in the tobacco blends from a Philip Morris USA database of a large number of commercial U.S. cigarettes were analyzed. MS smoke acetaldehyde is significantly correlated with "tar" yield and also with MS smoke carbon monoxide. These correlations are consistent with recently available data, including the 1999 Massachusetts Benchmark Study. MS smoke acetaldehyde yield is not correlated with reducing sugar (RS) concentration in the tobacco blend. For the available data over the time period 1985-1993, the concentration of reducing sugars (mean values for the brands tested) decreased. The correlations found between MS smoke acetaldehyde yield and "tar" and MS smoke carbon monoxide, and the percentage of the variance in AA yield explained by "tar" and MS smoke CO, coupled with the lack of correlation between "tar" normalized MS smoke AA and RS, plus related pyrolysis studies, support the hypothesis that MS smoke acetaldehyde yields are affected more by cigarette design characteristics influencing total smoke production than by tobacco reducing sugars. PMID- 12635007 TI - [Medicine and natural science in the profession of the diagnostic radiologist]. PMID- 12635008 TI - [Diagnostic radiology--let it drift or find new concepts?]. PMID- 12635010 TI - Use of iodine-based contrast media in digital full-field mammography--initial experience. AB - AIM: To investigate the use of iodine-based contrast media in digital full-field mammography. METHODS: After performing initial phantom studies, seven patients underwent digital mammography (Senographe 2000D, GE Medical Systems, Milwaukee, USA) using a specially filtered beam before as well as 60, 120, and 180 seconds after injection of 80 ml of iodine contrast medium (Ultravist 370, Schering AG, Germany). The precontrast mammograms were then subtracted from the postcontrast mammograms and the resulting images compared with a contrast-enhanced dynamic MRI study, performed on all women. RESULTS: Contrast medium accumulation within the tumors was visualized with a good quality in all cases. The conditions under which successful contrast-enhanced digital mammography can be performed were determined in phantom studies. CONCLUSIONS: Contrast-enhanced digital mammography has a potential for improving the visualization of breast tumors in mammography using special beam filtering, adjusted x-ray parameters, proper timing, and suitable subtraction software. PMID- 12635009 TI - [State of the art of MR colonoscopy]. AB - MR colonoscopy is suitable for colorectal cancer screening. This imaging modality is non-invasive, has no harmful side effects and provides a high diagnostic accuracy. MR colonoscopy has been found to be highly sensitive and specific in detecting colorectal lesions > 10 mm in size. Since colonic polyps are difficult to differentiate from residual stool, colonic cleansing has been unavoidable but reduces patient acceptance. Cleansing of the bowel can be avoided if the signal of stool is rendered different from its surrounding. This can be achieved by adding an agent to the food that tags the feces (fecal tagging). This article describes current techniques of MR colonoscopy as well as first results with fecal tagging. PMID- 12635011 TI - [Quality control of outpatient imaging examinations in North Rhine-Westphalia, Part II]. AB - PURPOSE: In the state of North Rhine-Westphalia (NRW), Germany, a survey was conducted on radiologic examinations ordered by general practitioners (GPs). Part II of this study aims to determine the quality of the process and outcome. The reference standard is the assessment of both radiologists and physicians without board certification in radiology working at a university hospital and in outpatient facilities. MATERIALS AND METHODS: AllGPs in NRW were asked to cooperate. Participating GPs filled out a questionnaire for each patient. The patients recorded the symptoms prompting the imaging examinations. The radiologists or other physicians performing the examinations were asked to provide the images and written reports and to complete a questionnaire. A file was created for each of the 394 patients with image documentation of at least one examination. Each file, which included medical history, physical findings, imaging documentation and written report, was sequentially forwarded to a board certified radiologist and to a physician without board certification in radiology working in a university hospital and in an outpatient facility. All physicians were requested to complete a structured questionnaire for each file. RESULTS: The referral diagnoses were rated as medically plausible in 81%, the indications for imaging found correct in 76%, the examination techniques considered appropriate in 69%, the clinical question answered in 63%, the interpretation judged medically correct in 50% and all incidental findings documented in 49%. In retrospect, 32 % of the examinations were judged superfluous. The sequence of multiple examinations performed on a particular patient was rated as appropriate in 51%. The interpretation revealed specialty-related differences. The plausibility of the referral diagnoses had a significant impact on the appropriateness of subsequent diagnostic investigations. Marked deficits showed sonography, performance by non-radiologists, self-referrals by GPs, gastroenterologic radiology and the ICD-10 coding (suspicion of cardiovascular disease). CONCLUSION: In the "best-case" scenario, the process quality proved to have moderate deficiencies and the outcome quality severe deficiencies. In consequence, GPs and radiologists should be more communicative by sharing information and exchanging opinions. GP self-referrals should be restricted. Sonography and examinations performed by physicians without board certification in radiology should undergo stricter quality controls. A more intensive interdisciplinary collaboration is needed to determine the optimum implementation of diagnostic imaging of gastroenterologic and cardiovascular diseases. PMID- 12635012 TI - [Recycling and reuse of disposable products in radiology]. AB - Legitimacy and appropriateness of recycling and reuse of single-use disposable medical devices is a common issue in many areas of medicine. Advocates of reprocessing claim economic savings and environmental benefits. From the manufacturers' point of view, the single use is a characteristic property of their products. Moreover, they warn that using of recycled instruments makes the procedure more difficult and increases the rate of adverse events. As comparative tests demonstrate, reused products are sometimes inferior to the genuine ones with respect to mechanical properties, but these differences may have relatively little consequence for the performance of the products and the success rate of the interventions. With rigorous cleaning and sterilizing of the instruments, patients do not run an increased risk of infection. Sterile single-use products are subject to the German Medical Products Act (1994) and the German Medical Devices Ordinance (1998). New legal provisions (in force since January 1st 2002) have restricted professional recycling of used devices further. Companies reprocessing medical products for the original users are free from certification liability. CE can no longer be issued for devices newly placed on the market. Lawyers controversially discuss whether patients must be informed about the use of recycled products. In view of the otherwise strict regulations concerning the information given to patients, it seems imperative to obtain a written informed consent when reprocessed devices are used. This might interfere with the confidential relationship of patients and physicians. The combination of increased risk, strict legal regulations, ethical concern and adverse publicity demand to refrain from reprocessing and reusing disposable equipment in diagnostic and interventional radiology. PMID- 12635013 TI - [Early results of ultrasound based calculation of broadband ultrasound attenuation and speed of sound in children and adolescents suffering from asthma]. AB - PURPOSE: To determine broadband ultrasound attenuation (BUA) and speed of sound (SOS) on the os caicis in asthmatic children. To correlate these findings with sex, age, weight and height, topical steroid intake, and asthma severity grade (ASG). PATIENTS AND METHODS: 178 children (ASG 1 - 3)/(98 m, 80 f; mean age 11.9 +/- 3.1 y) were consecutively chosen from 4/00 to 9/01. Children with any other chronic disease were excluded. BUA and SOS were measured using SAHARA (Hologic lnc. Waltham, USA). Regional normative BUA and SOS data of 3 299 children (obtained with the same system), were used to calculate age-, weight- and height matched standard-deviation-scores (SDS) for both sexes. Asthma severity grade and steroidal intake were determined. The highest topical steroid dosage was 500 micro g Fluticasone or 800 micro g Budesonide per day. RESULTS: 10/178 children were small and 7/178 tall per age (5.6 %/3.9 %), 11/178 children were light (6.2 %) and 9 heavy per age (5.0 %). 19 and 45 children had reduced BUA and SOS values, respectively. The following rates of reduced values were observed: girls: BUA 15.0 % (12/80), SOS 25.0 % (20/80); boys: BUA 7.1 %, SOS 25.5 % (7/98 and 25/98). Sexual differences were not significant. Reduced SOS-values were associated with higher severity and occurred significantly more frequent at children under steroidal intake (0.09 vs. 0.25 [BUA] and - 0.37 vs. - 0.07 [SOS]). CONCLUSION: Following our results an increase incidence of reduced speed of sound occurs in asthmatic children which is attributed to asthma severity and seems to be negatively influenced even by topically applied low dose steroids. This could be attributed to a steroid induced collagen synthesis deficiency followed by a reduced bone elasticity. Further studies, especially using a longitudinal study design are required to verify these findings. PMID- 12635014 TI - [Value of 14-gauge ultrasound-guided large-core needle biopsy of breast lesions: own results in comparison with the literature]. AB - PURPOSE: To report the false-negative rate, sensitivity, and specificity of 14 gauge ultrasound-guided large-core needle biopsy (14-G USSB) performed on breast lesions at our institution and, furthermore, to compare and discuss our own results with those reported in the literature. MATERIALS AND METHODS: This study includes 399 lesions examined by 14-G USSB. The results of the 14-G USSB were compared with the results of the surgical biopsy or, in cases of benign histology, were followed clinically. A key word search in two medical databases was undertaken to compare our data with those reported in the literature. The search was limited to the period from January 1990 to February 2002 and only original investigations published in English and German were included in our comparison. RESULTS: At our institution, 238 (59.6 %) benign and 161 (40.4 %) malignant lesions were diagnosed by 14-G USSB. The 14-G USSB was false-negative in 5 (1.25 %) of 399 cases. It has a sensitivity of 95.7 %, and specificity 100%. The literature reports false-negative rates between 0 % and 1.26 %, sensitivities between 86% and 100%, and specificities between 99.7 % and 100 %. On the basis of 3880 results from seven selected original papers and our own study, the false negative rate for 14-G USSB was calculated to be 0.4 % (16 of 3880). CONCLUSION: Based on our results and those reported in the literature, 14-G USSB can be considered safe and reliable in the assessment of breast lesions. PMID- 12635015 TI - [Diffusion-weighted MRI--a new parameter for advanced rectal carcinoma?]. AB - PURPOSE: To evaluate the predictive value of apparent diffusion coefficient (ADC) on therapy outcome of combined chemoradiation in patients with primary carcinoma of the rectum. MATERIALS AND METHOD: Prior to standardized, combined, neoadjuvant chemoradiation, 16 patients with primary carcinoma of the rectum (cT3) were examined with magnetic resonance imaging (MRI). Diffusion-weighted spin echo echo planar images (SE-EPI) and contrast-enhanced T 1 -weighted spin echo (SE) images at 1.5 Tesla were obtained. The mean ADC of the tumor region was calculated and correlated with the therapy outcome substantiated by postsurgical histopathologic staging. RESULTS: Tumor down-staging (pT0-2) occurred in 9 patients (therapy responders) and no down-staging (pT3) in 7 patients (therapy non-responders). The mean ADC measured 0.476 +/- 0.114 x 10(-3) mm 2/s in the responder group and 0.703 +/- 0.085 x 10(-3) mm 2/s in the non-responder group. Comparison of the mean ADC between the groups reached statistical significance (p = 0.001). CONCLUSION: The mean ADC might be a new quantitative parameter to predict therapy outcome of combined preoperative chemoradiation in patients with primary carcinoma of the rectum. PMID- 12635016 TI - [Improved evaluation of stenoses of hemodialysis fistulas by B-flow ultrasound]. AB - PURPOSE: To evaluate the new technique of B-flow ultrasound in assessing stenoses of hemodialysis fistulas. MATERIALS AND METHODS: 50 patients (mean age 58 years) with Brescia-Cimino-Shunts (27 shunts of the radial and 23 of the cubital artery) were prospectively assessed with intraarterial DSA and vascular ultrasound by independent examiners. Eligibility for the study was a shunt-volume of less than 400 ml/min and an angiographically suspected hemodynamically significant stenosis of the anastomosis or of shunt veins. Sonography was performed with a multifrequency ultrasound probe (5 to 10 MHz, Logic 700, GE) using B-mode, color coded Doppler sonography (CCDS) and B-flow technique. RESULTS: Anastomotic stenosis and stenosis of the shunt veins were equally distributed, found in 25 patients each. The measurements of the residual lumen of the 25 anastomotic stenoses were 1.47 to 3.43 mm (average: 2.3 mm) for intraarterial DSA, 1.57 to 3.73 mm (average: 2.6 mm) for B-mode ultrasound, 1.97 to 4.17 mm (average: 2.9 mm) for CCDS, 1.43 to 3.47 mm (average: 2.3 mm) for B-flow technique in the brightness mode and 1.6 to 3.47 mm (average: 2.4 mm) for B-flow technique in the B-mode. The brightness mode of the B-flow correlated best with intraarterial DSA (r=0.994), with a significantly lower correlation between CCDS and intraarterial DSA. B-flow displays less vascular distortion within the stenosis and fewer flow artifacts. Even in angulated stenoses, the detected intra- and poststenotic flow was markedly less angle-dependent in comparison with CCDS. B-flow clearly facilitates the visualization of hypoechoic plaques and intima proliferation. Furthermore, eccentric cicatricial stenoses, intima flaps or hypoechoic thrombi, which were not seen with DSA or B-mode, showed improved visualization in comparison with CCDS. CONCLUSION: Ultrasound B-flow makes it easier to assess the morphology and the local degree of stenotic hemodialysis fistulas. Flow detection is achieved with fewer artifacts and reduced angle dependence. This opens the way for better planning of interventional therapy. PMID- 12635017 TI - [Laser-induced thermotherapy (LITT) of lung metastases: description of a miniaturized applicator, optimization, and initial treatment of patients]. AB - PURPOSE: A thin-caliber applicator system was developed for introducing a laser fiber under CT guidance into lung metastases with only minimal complications. MATERIALS AND METHODS: A space-saving 5.5 French Teflon cannula with a titanium trocar and connectors for a laser light guide (2 or 3 cm Dornier Diffusor-Tip H 6111-T2 or H-6111-T3 coupled to a Dornier Medilas Fibertom 5100 laser, wavelength of 1064 nm) and a perfusion line for physiologic saline solution were developed. After puncture the laser Diffusor-Tip remains in the cannula and is cooled during its tissue passage by slowly flowing saline solution. The miniaturized applicator system (Monocath) was calibrated in nonperfused bovine liver for maximum energy supply and necessary flow of the cooling saline solution in reference to a commercially available 9 French laser catheter with an 11.5 French inducer sheath (Power-Applicator). The new applicator system was used for treating lung metastases in 10 patients over a period of 21 months. RESULTS: The size of heat coagulation in bovine liver was 24 +/- 2 ml using the miniaturized system with application of 15 W for 20 min and a saline flow of 0.75 ml/min, in comparison to a size of 29 +/- 7 ml for the commercial applicator (30 W, 20 min, 60 ml/min). All metastases could be safely approached with the miniaturized applicator, except for two metastatic lesions at the lung base in two patients. A minor pneumothorax developed in three patients and intrapulmonary bleeding in two. Contrast-enhanced CT demonstrated necrosis of the treated metastatic areas in 6 patients. Follow-up of three patients after 5, 6, and 8 months showed complete tumor regression with minimal scarring in one patient. CONCLUSION: The miniaturized applicator system enables the introduction of a laser fiber into pulmonary metastases with only minor complications. Complete ablation seems to be achievable in suitable patients with the applied laser energy and a slow cooling fluid flow rate. PMID- 12635018 TI - [Absorption of L-lysine diatrizoate from the gastrointestinal tract: the influence of surgery, inflammation and neoplasia]. AB - PURPOSE: To ascertain whether the absorption of L-lysine diatrizoate, a sodium free salt of the contrast-giving diatrizoic acid, from the gastrointestinal tract is increased by surgery, inflammation or neoplasia. MATERIAL AND METHODS: Using contrast medium containing L-lysine diatrizoate for intestinal opacification, this prospective study compared 32 radiographic examinations of the upper gastrointestinal tract with 52 radiographic examination of the lower gastrointestinal tract. In blood samples taken from the patients immediately after the radiographic examinations, the concentration of diatrizoic acid was determined by high pressure liquid chromatography. The results were correlated with sex, age, surgical history and any evidence of inflammatory or neoplastic diseases. RESULTS: The serum diatrizoic acid concentration in patients after oral administration was 3.62 (95% CI, 2.86 - 10.17) microg/ml. The titer was lower in patients who had undergone abdominal surgery than in patients without surgery. Serum diatrizoic acid concentration in patients after rectal administration was 0.30 (95% CI, 0.13 - 0.60) microg/ml. The titer was significantly higher (p < 0.05) in patients suffering from inflammatory conditions or neoplasms than in the other patients. CONCLUSION: The L-lysine salt of diatrizoic acid is absorbed in larger amounts from the upper than from the lower gastrointestinal tract. Absorption is not increased after abdominal surgery. However, inflammatory conditions and neoplasms of the large bowel increase the uptake of contrast medium from the intestine. PMID- 12635020 TI - [Compression-free biplanar radiography of the breast specimen with a specimen device]. AB - PURPOSE: To develop a device for specimen radiography of impalpable breast lesions with microcalcifications in order to perform compression-free radiography in two planes and to document the original orientation of the specimen on the radiographic film. MATERIALS AND METHODS: A curved plate with its radius equal to the film-focus-distance enables specimen radiography without compression and with minimizing artifacts induced by beam divergency. Adjustable markers can document the topographic orientation of the specimen on the radiographic film. RESULTS: The presented specimen device for biplanar radiography avoids interfering compression artifacts for both radiologist and pathologist. Documenting the original topographic orientation of the specimen on the film helps to guide the surgeon in cases requiring further resection. DISCUSSION: Biplanar specimen radiography without compression of the specimen measures the distance of microcalcifications to the margin of the resected tissue more accurately and thus provides a better basis for planning any necessary additional resection. PMID- 12635019 TI - [In-vitro comparison of the effectiveness of different high-speed rotatory catheters]. AB - OBJECTIVE: To determine the in vitro effectiveness of the Amplatz and Rotarex catheters for thrombus fragmentation and combined thrombus fragmentation and aspiration. MATERIALS AND METHODS: Clots (n = 100) from 5-day-old porcine blood (8.5 g) were fragmented with the 8F Rotarex trade mark catheter (RR; 40.000 rpm), which was directed with or without a coaxial guide wire (GW 0.020 inch [8 F RR(GW)], GW 0.014 inch [6 F RR(GW)]), and with the 7 F Amplatz catheter (ATD; 150.000 rpm), in an artery flow model (pulsed flow of 700 ml/min) simulating the superficial femoral artery. To increase the wall adherence of the thrombus, an additional equal number of silicon tubes (with an inner diameter of 7 mm) were fitted inside with a metallic mesh (wall-adherent thrombus [WAT]; 6 F RR(WAT), 8 F RR(WAT), 6 F R(WAT-GW), 8 F RR(WAT-GW), ATD(WAT) ). The effluent was passed through a three-step filter system (10 to 1000 micro m; pressure gradient 35 mmHg). RESULTS: Highest effectiveness was found for 8 F RR (no remaining thrombus detectable) compared to 6 F RR (0.08 g) and ATD (0.07 g), with a p < 0.001 for both methods. The aspirated volume was significantly lower when 6 F and 8 F RR were used with a coaxial guide wire (p < 0.001). The total emboli rate for 8 F RR was significantly lower compared to ATD (p < 0.001). Even when working without a guide wire, 8 F RR caused the lowest proportion of emboli (depending on the modification 0.12 % to 1.76 %; compared to all p < 0.05). Emboli rates reached 7.99 % for 6 F RR (compared to all p < 0.001) and were maximal 5.61 % for ATD. CONCLUSIONS: The tested mechanical thrombectomy catheters have a high potential for in vitro thrombectomy with moderate performance differences. In contrast to the guidewire-directed Rotarex fragmentation-aspiration device, the ATD trade mark fragmentation catheter and prototype Rotarex 6 F catheter have a considerable peripheral embolization rate of particles larger than 1000 micro m, probably requiring additional measures for in vivo thrombectomy. PMID- 12635021 TI - [A head-mounted display system for augmented reality: initial evaluation for interventional MRI]. AB - PURPOSE: To discuss the technical details of a head mounted display with an augmented reality (AR) system and to describe a first pre-clinical evaluation in interventional MRI. METHOD: The AR system consists of a video-see-through head mounted display (HMD), mounted with a mini video camera for tracking and a stereo pair of mini cameras that capture live images of the scene. The live video view of the phantom/patient is augmented with graphical representations of anatomical structures from MRI image data and is displayed on the HMD. The application of the AR system with interventional MRI was tested using a MRI data set of the head and a head phantom. RESULTS: The HMD enables the user to move around and observe the scene dynamically from various viewpoints. Within a short time the natural hand-eye coordination can easily be adapted to the slightly different view. The 3D perception is based on stereo and kinetic depth cues. A circular target with a diameter of 0.5 square centimeter was hit in 19 of 20 attempts. In a first evaluation the MRI image data augmented reality scene of a head phantom allowed good planning and precise simulation of a puncture. CONCLUSION: The HMD in combination with AR provides a direct, intuitive guidance for interventional MR procedures. PMID- 12635023 TI - [Iatrogenic aneurysma spurium of the pulmonary artery after insertion of Swan Ganz catheter: minimally invasive therapy using coil embolization]. PMID- 12635022 TI - [Primary lymphoma of the small intestine: MR diagnosis]. PMID- 12635024 TI - [Possibilities of diagnostic errors in space-occupying lesions of thoracic spinal canal]. PMID- 12635026 TI - [Severe allergic contact dermatitis with generalized spread due to bufexamac presenting as the "baboon" syndrome]. AB - HISTORY AND CLINICAL FINDINGS: A 48-year-old woman presented with acute, pruritic, sharply demarcated, erythematous, maculopapulous exanthem in the anogenital area with disseminated maculae over the back. Several days before, the patient had applied as topical treatment a bufexamac-containing ointment to the anal region. INVESTIGATIONS: The patch test showed an allergic test reaction to bufexamac. DIAGNOSIS: The case presents a serious allergic contact dermatitis with generalization, imitating a baboon syndrome, unequivocally linked to the previous topical treatment. CONCLUSION: During the last 10 years allergic reactions to bufexamac have increasingly been reported, sometimes with erythema multiforme-like reactions. Because of the high rate of sensitization, the serious clinical course of bufexamac allergy and the insidious symptoms of this side effect, sometimes mimicking the disease to be treated, the substance should be used neither for proctological nor for dermatological diseases, even more as these patients are considered to be at high risk of developing allergic contact dermatitis because of the abnormal skin barrier. Considering the data presented, the use of bufexamac should be critically reassessed. PMID- 12635025 TI - [Lowering of blood pressure, blood pressure amplitude and heart rate by treatment with valsartan or valsartan/hydrochlorothiazide. Results of an open observation study of 11,447 hypertensives]. AB - OBJECTIVE: This trial investigated the decrease of blood pressure, safety and tolerability of valsartan, an angiotensin-receptor blocker, alone or combined with the diuretic hydrochlorothiazide. In addition, the effect on pulse pressure, heart rate and mean blood pressure were recorded as independent markers of cardiovascular risk. PATIENTS AND METHODS: 12278 hypertensive patients were treated for 12 weeks with valsartan alone or combined with hydrochlorothiazide in a multi-centre open trial. Previously established antihypertensive treatment was continued. Systolic and diastolic pressure, pulse pressure and resting heart rate were recorded at the start of the trial and at 1, 4 and 12 weeks thereafter. RESULTS: Data on 11447 were complete enough to be evaluated (54% males, 46% females; mean age 60.8 years). 44% of the patients were at the same time receiving other antihypertensive drugs at the start of the trial, but the numbers so treated decreased during the trial period. Systolic pressure was reduced by 27.8 mmHg. diastolic pressure by 14.7 mmHg, with a calculated reduction of pulse pressure from 71.5 to 58.7 mmHg. Heart rate was reduced by 3.4 beats. The treatment had to be stopped prematurely in 2.3% of patients because of side effects. There were no severe or life-threatening side effects. CONCLUSION: Treatment with valsartan alone or combined with hydrochlorothiazide provides effective blood pressure reduction and is well tolerated. This trial suggests that, in addition to the fall in blood pressure, the independent risk factors of cardiovascular disease, pulse pressure and resting heart rate, were also reduced during administration of valsartan. PMID- 12635027 TI - [Adoptive transfer of Epstein-Barr virus-specific T-lymphocytes in chronic active Epstein-Barr infection]. AB - HISTORY: A 27-year-old man was admitted because of intermittent fever, fatigue, lymphadenopathy and splenomegaly for 20 years. Chronic administration of 6 - 8 g aspirin per day (self-prescribed) resulted in limited control of symptoms and in the development of analgesic nephropathy. INVESTIGATIONS: The patient had prominent splenomegaly and lymphadenopathy without histological signs of malignancy. Monocytosis and T-lymphopenia were also present. Infectious disease testing revealed IgG+/IgM- EBV serology and EA-EBV-mRNA nested PCR clearly demonstrated the presence of lytic EBV-proteins in PBMCs. TREATMENT AND COURSE: As chronic active Epstein-Barr virus infection (CAEBV) was highly probable, treatment with aciclovir, gancilovir and steroids was started. Because treatment failed adoptive T-cell transfer with autologous EBV-specific T-cells was performed. After three consecutive infusions the patient responded with a complete remission of fever, fatigue, lymphadenopathy and splenomegaly without adverse effects. Retrospective real-time PCR analysis showed a decrease in viral load from 62847 copies/ microg DNA to 45 - 250 copies after treatment. The patient remains in stable remission without signs of CAEBV (> 4 years). CONCLUSION: Adoptive transfer of autologous, EBV-specific T-lymphocytes is a promising treatment in CAEBV. PMID- 12635028 TI - [RNA interference--a new biological mechanism with significance for gene therapy]. PMID- 12635030 TI - [Isolated noncompaction of ventricular myocardium syndrome--a rare structural heart disease]. PMID- 12635029 TI - [Diagnosis of adrenal cortex insufficiency]. PMID- 12635031 TI - [DRG's: diagnoses rewarded, not therapy?]. PMID- 12635032 TI - [Value of full heparinization in venoatrial acute thromboembolic insult]. PMID- 12635033 TI - [What significance does the blood cell sedimentation rate have?]. PMID- 12635034 TI - [Prevention using vitamins and trace elements]. PMID- 12635035 TI - [Polyneuropathy--therapy]. PMID- 12635036 TI - [Which studies, which defects?]. PMID- 12635038 TI - [Predicting outcomes of burned patients]. PMID- 12635039 TI - [Prehospital use of hypertonic solutions for traumatic brain injury]. AB - Hypertonic solutions efficaciously lower increased intracranial pressure and improve cerebral hemodynamics in particular at the level of microcirculation. Therefore hypertonic solutions have been introduced to the in-hospital intensive care treatment of brain-injured patients. In view of the disturbed cerebral perfusion and oxygenation in the acute phase following traumatic brain injury, early initiation of treatment is desirable to improve long-term outcome and to minimize secondary brain damage. Two different hypertonic isooncotic solutions are licensed in Germany for prehospital and in-hospital treatment of acute hypotension due to hypovolaemia and haemorrhagic shock. Both solutions offer interesting therapeutic options for treatment of patients with increased intracranial pressure. Limitation of license to the treatment of shock in Germany does not allow routine prehospital use in isolated head trauma at the moment. In other European countries, the approval for the prehospital use of hypertonic isooncotic solutions has been modified. PMID- 12635040 TI - [Integration of preexisting diseases and risk factors in the Abbreviated Burn Severity Index (ABSI)]. AB - OBJECTIVE: There is no doubt that underlying medical problems such as concomitant diseases or risk factors play a role in increasing patient morbidity and mortality. These factors are already integrated in trauma scores but preexisting diseases have no impact on burn scores yet. This study was performed to examine the predictive value of the classical burn variables that are integrated in the Abbreviated Burn Severity Index (ABSI). The preexisting diseases and risk factors in burn patients within our burn center were evaluated, with the aim of incorporating these evaluations into a new burn score. This modified burn score was used to optimize the predictive value of burn mortality. METHODS: This study included 443 intensive care burn patients. Demographic, injury, age, total body surface area burned (TBSAB), full thickness burn (FTB), inhalation injury (IHT), sex, medical comorbidities, intensive care and outcome data were documented. Univariate analyses, stepwise logistic regression and the Receiver Operating Curve were used to generate values for the probability of death. RESULTS: Univariate analyses identified the following risk factors for their relationship with mortality: TBSAB, age, IHT, FTB, sex and medical comorbidities (cardiovascular, pulmonary, renal and endocrinological). Logistic Regression showed that total body surface area burned and age correlated most significantly with the probability of poor outcome. There were weaker correlations between IHT and FTB. No main effect was registered for gender and preexisting medical problems. The greatest area under the ROC curve was registered for our modified ABSI when comorbidities and risk factors were integrated. CONCLUSION: The results of this study show that the Abbreviated Burn Severity Index is an appropriate burn score for estimating the risk of mortality after burn trauma. However, in addition to the classical variables, preexisting diseases and risk factors have a significant influence on the outcome and therefore should be incorporated into a new burn score to predict mortality more accurately. PMID- 12635041 TI - [Point of care (POC) monitoring in anesthesia and intensive care--an overview of available POC systems]. AB - There is an increasing trend to have data rapidly ready at the patient's bedside. The general principle behind point-of-care (POC) testing is that "faster is better" ('from vein to brain'). The entire process for laboratory testing includes withdrawal of blood into special (pre-labelled) tubes, transportation of the sample to the central laboratory where the plasma is separated from the blood cells by centrifugation, carefully pippeting of a defined volume of plasma that is placed in the analyzer. POC instruments provide us with the potential to do old things in new ways. With the help of portable POC analyzers a variety of laboratory parameters including coagulation parameters, blood gas analysis, electrolytes, markers of organ function can be measured next to the patient's bed thus significantly shorting turnaround-time (TAT). Cost analyses of new monitoring devices are necessary in today's climate of cost savings. It is important to capture all costs and not only costs of the test kits. Direct costs (e. g. test cartridges, costs for the analyzers, cost for quality control) may constitute only a small percentage of the true costs. Hidden costs consist of overhead costs (e. g. transportation) and the consequences of delayed results. The present overview summarizes the available POC systems in Germany and may serve as a decision maker for those who are interested in introducing POC monitoring systems. PMID- 12635042 TI - [Mydriasis not reacting to light during an uneventful esophagectomy--drug-related side effect of Akrinor?]. AB - We present a 72 year old patient who underwent an esophagectomy uneventfully. At the end of the procedure the anaesthetist diagnosed a mydriasis not reacting to light. The mydriasis lasted for 3 hours. 30 minutes after diagnosis a cranial computed tomography scan showed no abnormalities. The tracheal tube was removed after 16 h of artificial ventilation, the patient was awake and responded to questions. The neurological examination was normal, as well as the control CT scan. With regard to a mydriasis the following differential diagnosis have to be considered: cerebral ischemia, cerebral hematoma, metabolic encephalopathy, shock, and a drug side effect. We state that an alpha-mimetic effect of the drug mixture Akrinor, the patient had received shortly before diagnosis of the mydriasis, is the most likely explanation. One 2 mls syringe contents of 200 mg norephedrine-ethyl-theophylline and 10 mg noradrenaline-ethyl-theophylline. PMID- 12635043 TI - [Thoracic epidural anaesthesia]. PMID- 12635044 TI - Emergency care in the new millennium: current developments and perspectives. PMID- 12635046 TI - [Ketamine in emergency care: new standard or exclusive alternative?]. PMID- 12635047 TI - [Preclinical care of polytrauma: rush or stay?]. PMID- 12635048 TI - [Reanimation in the new millennium--new standard or proven treatment?]. PMID- 12635049 TI - [Small volume resuscitation--solution for the routine or ultima ratio?]. PMID- 12635050 TI - [Avalanche accident--ways to safety or hopeless case]. PMID- 12635051 TI - [Dangers in diving and typical diving accidents]. PMID- 12635052 TI - S100beta and NSE: stroke surrogate signals or natal neural noise? PMID- 12635053 TI - Are serum S100beta proteins and neuron-specific enolase predictors of cerebral damage in cardiovascular surgery? AB - OBJECTIVE: To examine whether serum concentrations of S100beta protein and neuron specific enolase (NSE) are predictors of cerebral damage in cardiovascular surgery. DESIGN: Prospective clinical study. SETTING: University hospital. PARTICIPANTS: Eighteen patients with conventional cardiopulmonary bypass (CPB), 7 with selective cerebral perfusion (SCP), and 3 volunteers (blood samples). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: S100beta and NSE were measured in the blood obtained at 7 time points during and after operation. The concentrations of these markers in the blood from the surgical field and the cell saver device, and the influence of graded hemolysis (in vitro) on the concentrations of these proteins were also examined. The mean values of S100beta in the CPB group (2.08 +/- 2.00 ng/mL) and the SCP group (1.46 +/-0.77 ng/mL) were highest after aortic declamping and after termination of SCP, respectively. The mean values of NSE in the CPB group (29.1 +/- 14.0 ng/mL) and the SCP group (31.2 +/- 13.6 ng/mL) were highest after termination of CPB and at the end of the operation, respectively. Three patients suffered from cerebral complications, but the elevation of these markers during operation was indistinguishable from those in the other patients. Peak concentrations of S100beta protein in the CPB group and NSE in the SCP group were correlated with the duration of aortic cross clamping and CPB, respectively. S100beta protein and NSE concentrations in the blood from the surgical field were significantly larger than those in arterial blood, whereas the concentrations in the blood in the cell-saving device were not elevated. The concentration of S100beta protein was not influenced by the extent of hemolysis, whereas NSE concentration was markedly elevated by hemolysis. CONCLUSION: A large part of the increases in S100beta protein and NSE during CPB and SCP is not attributed to neuronal damage, but to contamination with the blood from the surgical field. To determine whether these markers are useful to predict neurologic complications, it will be necessary to exclude contamination from the surgical field as observed in the present study. PMID- 12635054 TI - Association of sex with perioperative mortality and morbidity after carotid endarterectomy for asymptomatic carotid stenosis. AB - OBJECTIVE: To examine whether perioperative morbidity and mortality after carotid endarterectomy depend on the sex and the presence of symptoms on presentation. DESIGN: Retrospective review of quality assurance database prospectively collected. SETTING: A university teaching hospital. PARTICIPANTS: One thousand two hundred eighty-seven patients who had 1,503 carotid endarterectomies from 1990 to 1999 from a quality assurance database. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The cases were divided into 4 groups by sex and symptoms on presentation: male-symptomatic (MS), male-asymptomatic (MA), female-symptomatic (FS), and female-asymptomatic (FA). The 4 groups were compared for preoperative demographic and comorbidity profiles, carotid plaque characteristics, and outcome. Outcome measures included in-hospital stroke, myocardial infarction (MI), congestive heart failure (CHF), and death. There were 496 cases in the MS group, 407 in the MA group, 315 in the FS group, and 285 in the FA group. Women were less likely to have a history of coronary artery disease, prior MI, or smoking, and their carotid plaques were less likely to be ulcerated or contain intraplaque hemorrhage. Even when controlling for the comorbidities and plaque characteristics, the incidence of each of the complications examined was low and not significantly different between the sexes in both the symptomatic and asymptomatic groups. The rate of stroke or death was 3.0% (MS) versus 1.9% (FS) (p = NS) and 1.2% (MA) versus 1.8% (FA) (p = NS). CONCLUSION: There is no significant sex difference in perioperative cardiac or cerebrovascular complications. Women with symptomatic or asymptomatic carotid stenosis can have acceptably low complication rates from carotid endarterectomy and may benefit from the surgery as much as men. PMID- 12635055 TI - A case series of low-dose fenoldopam in seventy cardiac surgical patients at increased risk of renal dysfunction. AB - OBJECTIVE: To evaluate the usefulness of low-dose fenoldopam mesylate in patients at risk of developing renal dysfunction after cardiac surgery requiring cardiopulmonary bypass. DESIGN: A prospective, single-center, observational study. SETTING: University teaching hospital. PARTICIPANTS: Seventy patients scheduled for elective cardiac surgery with one or more predefined risk factors for renal dysfunction. INTERVENTIONS: After induction of anesthesia, fenoldopam (0.03 microg/kg/min) was administered throughout surgery and into the postoperative period, until the patient was stable and weaned from all other vasoactive agents. Perioperatively, fenoldopam was also used as a second-line antihypertensive agent as required. MEASUREMENTS AND MAIN RESULTS: No patient developed renal failure that required dialysis, whereas 7.1% (5/70) developed non dialysis-dependent renal dysfunction. Four out of these 5 patients had 2 or more risk factors (9.5%). Higher preoperative creatinine levels, a history of hypertension, myocardial infarction within 5 days of surgery, and a preoperative diagnosis of chronic renal insufficiency were all good predictors of postoperative non-dialysis-dependent renal dysfunction. Discharge serum creatinine levels were lower than preoperative levels (1.16 +/- 0.36 mg/dL v 1.26 +/- 0.34 mg/dL, p < 0.05). CONCLUSION: These findings suggest that renal function was preserved in patients at increased risk for renal dysfunction after cardiac surgery when low-dose fenoldopam was used in the perioperative period. However, a randomized, controlled trial is required to establish efficacy. PMID- 12635056 TI - Ventilator-associated pneumonia: Incidence, risk factors, outcome, and microbiology. AB - OBJECTIVE: To determine the incidence, risk factors, outcome, and pathogens of ventilator-associated pneumonia (VAP) in a cardiac surgical intensive care unit (ICU). DESIGN: Prospective study. SETTING: Escorts Heart Institute and Research Centre, New Delhi, India. PARTICIPANTS: Nine hundred fifty-two consecutive patients undergoing cardiac operations who received intermittent positive pressure ventilation (IPPV). INTERVENTIONS: All patients were assigned into VAP (n = 25) and non-VAP (n = 927) groups. MEASUREMENTS AND MAIN RESULTS: Risk factors and other variables were analyzed with univariate and multivariate analysis. Of the 952 patients studied, 25 (2.6%) had VAP. On univariate analysis, significant risk factors were emergency surgery, chronic obstructive pulmonary disease (COPD), reintubation, coma, steroid treatment, intra-aortic balloon counterpulsation (IABC), enteral feedings, tracheostomy, acute physiology, age, and chronic health evaluation (APACHE II) score, prior antibiotics, and IPPV hours. On multivariate analysis, IPPV hours (153.75 +/- 114.44 v 19.65 +/- 7.99; p < 0.001) and steroids (20% v 0%; p < 0.001) were independent predictors of VAP. The most common pathogens isolated were Pseudomonas aeruginosa (22), Escherichia coli (10), Klebsiella pneumoniae (4), Staphylococcus species (4), and Acinetobacter species (2). The mortality rate in VAP was 16% as compared with 0.2% in non-VAP cases (p < 0.001). CONCLUSION: These data suggest that by univariate analysis the risk factors for VAP were emergency surgery, COPD, reintubation, coma, steroid treatment, IABC, enteral feedings, tracheostomy, APACHE II score, prior antibiotics, and IPPV hours. On multivariate analysis, only IPPV hours and steroids were independent predictors of VAP. Pseudomonas aeruginosa is the most common pathogen associated with VAP, and the mortality is increased with VAP. PMID- 12635057 TI - Is very early extubation after lung transplantation feasible? AB - OBJECTIVE: To evaluate donor graft function, intraoperative blood consumption, and oxygenation and hemodynamic stability in patients undergoing lung transplantation. DESIGN: Prospective pilot study. SETTING: University hospital. PARTICIPANTS: Forty-three patients undergoing lung transplantation from January 1999 to June 2001. INTERVENTIONS: Hemodynamic monitoring, early extubation, and noninvasive ventilation criteria. MEASUREMENTS AND MAIN RESULTS: The 31 nonearly extubated patients showed a lower PaO(2)/fraction of inspired oxygen (F(I)O(2)), a higher mean pulmonary arterial pressure, extravascular lung-water index (EVLWI) and vasoactive drug support (norepinephrine), and more blood products consumption than 12 early extubated patients at the end of surgery. Seven of 12 early extubated patients did not show any signs of respiratory failure after tracheal extubation; they were alert and able to perform deep breathing exercise and coughing. In the other 5 patients, hypoxemia, hypercapnia, and an increase of respiratory rate >30 breaths/min were observed. The intermittent application of noninvasive pressure ventilation by face mask avoided endotracheal intubation. CONCLUSION: The use of a short-acting anesthetic drug, appropriate intraoperative extubation criteria, epidural analgesia, and postoperative noninvasive ventilation make early extubation of lung-transplanted patients possible and effective. PMID- 12635058 TI - Early extubation after single-lung transplantation: analysis of the first 106 cases. AB - OBJECTIVE: To determine if a modern anesthetic approach permits extubation immediately after surgery for single-lung transplantation. DESIGN: A retrospective study of all patients undergoing single-lung transplantation from June 1993 to December 1999 in Denmark. SETTING: Rigshospitalet, Copenhagen University hospital. PARTICIPANTS: One hundred six consecutive patients scheduled for single-lung transplantation. INTERVENTIONS: From July 1997, the anesthetic approach was changed to facilitate early extubation. The changes included epidural analgesia and short-acting anesthetic drugs. MEASUREMENTS AND MAIN RESULTS: One hundred six patients were anesthetized for single-lung transplantation. The first 33 patients were moved to the intensive care unit for postoperative mechanical ventilation. After the change of anesthesia technique, 53 of 73 patients were extubated in the operating room. Eleven patients needed reintubation within the first 24 hours because of respiratory insufficiency, pulmonary edema, hemorrhage, or pneumothorax. The need for reintubation increased the length of stay in the intensive care unit by 1 day from 2 to 3 days (NS). The possibility of early extubation or the need for reintubation was not related to age, weight, sex, preoperative condition, mode of transport of the graft, duration of graft ischemia, or side of transplantation. CONCLUSION: This study has shown that it is possible to extubate patients in the operating room immediately after single-lung transplantation in the majority of cases. PMID- 12635059 TI - The effect of high-frequency ventilation of the lungs on postbypass oxygenation: A comparison with other ventilation methods applied during cardiopulmonary bypass. AB - OBJECTIVE: To compare the effect of high-frequency ventilation versus other ventilation methods applied during cardiopulmonary bypass on postbypass oxygenation. DESIGN: Prospective, randomized study. SETTING: University hospital. PARTICIPANTS: Seventy-five patients undergoing coronary artery bypass graft surgery. INTERVENTIONS: Patients were allocated to 5 equal groups of different ventilation methods during bypass. Groups 1 and 2 received high-frequency, low volume ventilation with 100% and 21% oxygen, respectively. Groups 3 and 4 received 5 cm H(2)O of continuous positive airway pressure (CPAP) with either 100% or 21% oxygen. Patients from group 5 were disconnected from the ventilator during the bypass period. MEASUREMENTS AND MAIN RESULTS: Spirometry data, blood gas analysis, oxygen saturation as measured by pulse oximetry, and end-tidal carbon dioxide were recorded 5 minutes before chest opening, 5 minutes before bypass, 5 minutes after bypass, 5 minutes after chest closure and 6, 12, 18, and 24 hours after surgery. There were no differences in compliance and mean airway pressures. Alveolar-to-arterial oxygen gradients increased, and PaO(2) decreased significantly (p < 0.05) in all groups 5 minutes after bypass and this trend continued in the postoperative period. Patients from group 3 had higher PaO(2) and lower alveolar-to-arterial oxygen gradients, 5 minutes after weaning from bypass (p < 0.05). Extubation times were similar in all groups. CONCLUSIONS: The alveolar-arterial oxygen gradient was lower, and the PaO(2) was higher 5 minutes after bypass in patients receiving CPAP (100% O(2)) as compared with those ventilated with high-frequency ventilation. PMID- 12635060 TI - Effects of positive-pressure ventilation, pericardial effusion, and cardiac tamponade on respiratory variation in transmitral flow velocities. AB - OBJECTIVE: To determine the effects of positive-pressure ventilation and experimentally induced pericardial effusion and tamponade on transmitral flow velocities in dogs. DESIGN: Descriptive. SETTING: University laboratory. PARTICIPANTS: Eleven tracheally intubated and mechanically ventilated dogs. INTERVENTIONS: Experimental pericardial effusion and cardiac tamponade were created by pericardial injection of warm saline. MEASUREMENTS AND MAIN RESULTS: Hemodynamic parameters and pericardial pressures were monitored in the 11 dogs. Pulsed-wave Doppler tracings of mitral valve flow were obtained at the leaflet tips along with hemodynamic measurements at 4 stages: control, effusion (no decrease in mean arterial pressure), tamponade (>or=40% decrease in mean arterial pressure), and tamponade relief (after evacuation of pericardial fluid). Maximal variation (36%) in transmitral flow velocity over the respiratory cycle during positive-pressure ventilation was seen in the control stage. In the effusion and tamponade stages, variation in transmitral flow velocity decreased progressively to 29% (p = 0.1804, not significant) and 16% (p < 0.0001), respectively. CONCLUSION: Intrathoracic pressure and lung volume changes caused by positive pressure ventilation influence transmitral flow velocity patterns. Respiratory variation in transvalvular flow is pronounced during standard positive-pressure mechanical ventilation, decreases in the presence of pericardial effusion, and becomes almost nonexistent when cardiac tamponade is present. These findings show that the echocardiographic criteria used to diagnose cardiac tamponade based on mitral valve inflow patterns are different during positive-pressure ventilation from spontaneously breathing subjects. PMID- 12635061 TI - Comparison of three remifentanil dose-finding regimens for coronary artery surgery. AB - OBJECTIVES: To identify the remifentanil dosing regimen providing safe and optimal anesthetic conditions during coronary artery bypass graft surgery and to evaluate postoperative recovery characteristics. DESIGN: Open-label, randomized, parallel group. SETTING: Three centers in the United States. PARTICIPANTS: Seventy-two patients with left ventricular stroke volumes >or=50 mL. INTERVENTIONS: Patients were randomized to remifentanil doses of 1 microg/kg/min (group 1, n = 23); 2 microg/kg/min (group 2, n = 24), or 3 microg/kg/min (group 3, n = 25). Somatic, sympathetic, and hemodynamic responses indicating inadequate anesthesia were treated with bolus doses of remifentanil, 1 to 2 microg/kg, and infusion rate increases, and, if necessary, isoflurane 0.5% to 1.0% was added as a rescue anesthetic. In the intensive care unit, the remifentanil infusion was reset to 1 microg/kg/min, with midazolam administered for supplemental sedation and morphine for analgesia. MEASUREMENTS AND MAIN RESULTS: The durations of anesthesia, surgery, and cardiopulmonary bypass were similar for the 3 study groups. In addition, dose of lorazepam premedication, time to loss of consciousness, preoperative left ventricular ejection fraction, age, weight, and sex were similar for the 3 study groups. Remifentanil alone (infusion and boluses) prevented and controlled all responses to stimulation in 44% of group 3, 37% of group 2 and 9% of group 1 patients intraoperatively. Isoflurane (0.5%-1% inspired) rescue was successful in the remaining patients in each group. Hypotension indicating discontinuation of isoflurane and reductions of remifentanil infusion rates occurred in 64% to 75% of all patients. The optimal range of remifentanil infusion was 2 to 4 microg/kg/min with isoflurane to supplement the opioid. Fifty-one patients (71%) met the criteria for extubation within 6 hours postoperatively; because of surgical practice differences, only 30 patients (59%) were actually extubated. CONCLUSIONS: After lorazepam premedication, remifentanil infusion (2-4 microg/kg/min) supplemented intermittently with low inspired concentrations of isoflurane provided an effective anesthetic regimen for coronary artery bypass graft surgery. Early extubation times were feasible after remifentanil continuous infusions (1-5 microg/kg/min) used as the primary anesthetic component intraoperatively and for analgesia ( 0.05) different from that of females. In contrast, the valve movement responses of V. angasi to U were size- and age dependent; smaller and younger individuals (median shell length = 36.8 mm; median age = 0.7 years) were 22% more sensitive (p 2.5 mmol/L provided the highest sensitivity (0.62) and specificity (1.00) for MITO disease. A baseline and +1 min sample provided optimal sensitivity and specificity for GSD and AMPD deficiency. Catheter placement in any vein other than the ipsilateral antecubital resulted in attenuated lactate responses (P < 0.0001). A pre-FIT sugar bolus did not alter the postexercise lactate or ammonia response. Thus, a modified FIT was helpful in the diagnosis of GSD and excluding AMPD deficiency, but not in the diagnosis of MITO disease. Catheter placement is critical to the interpretation of a FIT, whereas pretesting diet is less important. PMID- 12635124 TI - Optimal distance for segmental nerve conduction studies revisited. AB - In 1972, Maynard and Stolov showed that the experimental error in calculating nerve conduction velocity (NCV) depends on errors of latency and distance measurements. Their data suggested that a minimum distance of 10 cm should be used when calculating NCV because of an increase in error >/= 25% at shorter distances. The object of this study was to reestablish the minimum distance using current technology. Twenty physicians measured the proximal and distal onset latencies of the same stored ulnar compound muscle action potentials, as well as the forearm skin distance. The mean and standard deviation of the errors in conduction time and distance were determined. A spreadsheet was constructed, determining the error in NCV for a variety of distances and NCVs. The average conduction time between proximal and distal stimulation was 3.23 ms, with a standard deviation of 0.09 ms. The mean distance measurement was 212.6 +/- 2.1 mm. The errors in NCV were significantly less than previously reported. The experimental error increased as distance decreased, reaching 20% with distances less than 6 cm. The latency error accounted for 71% of the total experimental error, as opposed to 90% in the previous study. With advances in electrodiagnostic equipment, NCV can reasonably be calculated at distances less than 10 cm, perhaps as low as 5-6 cm. PMID- 12635125 TI - Nerve growth factor expression in human dystrophic muscles. AB - Nerve growth factor (NGF) is a neurotrophin that is expressed during muscle development and is also capable of favoring muscle regeneration in experimental studies. The presence of NGF in muscular dystrophies, such as Duchenne and Becker muscular dystrophies, has never been fully explored. By means of immunohistochemistry, we show that regenerating muscle fibers from such patients consistently express NGF, as do myofibroblasts and mast cells. By contrast, rest fibers from dystrophic patients, as well as muscle fibers from healthy, control patients and even regenerative muscle fibers in polymyositis do not show NGF immunoreactivity. The paracrine effect of NGF on muscle regeneration, as well as its chemoattractant capacities for mast cells, may contribute to explaining why regenerating fibers most frequently occur in clusters and why mast cells are more numerous in dystrophic muscles. Moreover, being a mediator of wound healing and tissue fibrosis, NGF may contribute to long-term muscle regeneration impairment by tissue fibrosis in the muscular dystrophies. PMID- 12635126 TI - Acute compartment syndrome of the leg following diagnostic electromyography. AB - Acute compartment syndrome of the lower extremity is typically associated with some form of trauma, either fracture or blunt injury. Accurate diagnosis and urgent treatment of this condition is required for preservation of the viability of the limb. We report the case of a 73-year-old man who developed an acute compartment syndrome of the leg after diagnostic electromyography. Potential causes and treatment are discussed. PMID- 12635127 TI - Literature review: nervepace digital electroneurometer in the diagnosis of carpal tunnel syndrome. AB - Over the last two decades, several diagnostic devices have been developed to assess patients with suspected carpal tunnel syndrome (CTS). One such device is the Nervepace Digital Electroneurometer (NDE). At this time, however, the AAEM concludes that the current literature does not support the substitution of the NDE for standard electrodiagnostic studies in the clinical evaluation of patients with CTS. PMID- 12635128 TI - The chin reflex. PMID- 12635130 TI - The clinical evaluation of HER-2 status: which test to use? AB - Accurate determination of the status of the type I receptor tyrosine kinase HER-2 in breast carcinomas provides significant insight into patient prognosis and may also inform selection of chemotherapeutic and hormonal treatments. At present, however, the single most important application of HER-2 testing is in the selection of patients for treatment with targeted therapies such as Herceptin. Although, based on current literature, fluorescence in situ hybridization (FISH) detection of HER-2 gene amplification may provide more accurate information in this context, this method is not yet widely available. Therefore, screening by immunohistochemistry (IHC) for HER-2 protein, backed by rigorous quality controls and FISH testing of equivocal cases with intermediate staining intensity, remains the current practice. In laboratories with highly standardized testing and quality assurance procedures, this protocol appears highly effective. Improvements in fixation procedures, standardization of antibodies, and use of automated image analysis may all increase the precision of IHC testing. However, on the basis of current data, there is a case to be made for the wider implementation of FISH testing to determine HER-2 status in breast cancer. PMID- 12635131 TI - Correlation between immunohistochemistry (HercepTest) and fluorescence in situ hybridization (FISH) for HER-2 in 426 breast carcinomas from 37 centres. AB - Accurate diagnostic assessment of HER-2 is essential for the appropriate application of the humanized anti-HER-2 monoclonal antibody trastuzumab (Herceptin) to the treatment of patients with metastatic breast cancer. The diagnostic test needs to be applicable to archival, fixed tissue removed at excision, in many cases several years earlier. We compared the assessment of HER 2 by immunohistochemistry (IHC; HercepTest) and fluorescence in situ hybridization (FISH) in 426 breast carcinomas from patients being considered for trastuzumab therapy. The tumours were tested in three reference centres having been sent in from 37 hospitals. Only 2/270 (0.7%) IHC 0/1+ tumours were FISH positive. Six of 102 (5.9%) IHC 3+ tumours were FISH negative. Five of the six had between 1.75 and 2.0 HER-2 gene copies per chromosome 17 and the sixth had multiple copies of chromosome 17. Thirteen per cent of tumours were IHC 2+ and overall 48% of these were FISH positive but this proportion varied markedly between the centres. Sixty IHC-stained slides selected to be enriched with 2+ cases were circulated between the three laboratories and scored. There were 20 cases in which there was some discordance in scoring. Consideration of the FISH score in these cases led to concordance in the designation of positivity/negativity in 19 of these 20 cases. These data support an algorithm in which FISH testing is restricted to IHC 2+ tumours in reference centres. The results may not extrapolate to laboratories with less experience or using different methodologies. PMID- 12635132 TI - Expression of cyclins E, A, and B, and prognosis in lymph node-negative breast cancer. AB - Unexpected outcomes in breast cancer demand a refinement of prognostic criteria. This study therefore investigated the prognostic relevance of cyclin expression in a cohort of 332 T1-T2 N0 infiltrating ductal carcinomas with long-term follow up (median 99 months). By univariate analysis, tumour size, histopathological grade, hormone receptor content, cyclin E, cyclin B, and the Ki-S5 (Ki-67) index significantly predicted disease-specific and metastasis-free survival. Cyclin A did not achieve statistical significance. In a multivariate analysis, both cyclin E [relative risk (RR) 2.01, p = 0.021] and cyclin B (RR 1.85, p = 0.033) were selected as independent prognosticators of metastasis-free survival when the Ki 67 index was omitted, but only cyclin E expression was associated with disease specific survival (RR 2.56, p = 0.006). When Ki-67 was included as a covariate, cyclin E lost its significance with respect to disease-specific survival but remained significant for metastasis-free survival. In an analogous analysis including Ki-67, the number of concurrently overexpressed cyclins did not attain statistical significance regarding disease-specific survival but was selected as the leading predictor of metastatic disease. It is concluded that combined overexpression of cyclins may imply genetic instability enhancing metastatic potential, but that survival ultimately depends on the proliferative activity of tumour cells. PMID- 12635133 TI - Maspin expression in invasive breast cancer: association with other prognostic factors. AB - Maspin is a unique serine protease inhibitor with a molecular weight of 42 kDa. It has been shown to inhibit tumour cell motility and invasion in cell culture, and tumour growth and metastasis in animal models. There is very limited data on the prognostic utility of maspin in human breast cancer. We performed a preliminary study to assess the associations of maspin with other established prognostic factors in invasive breast cancer (IBC). 1068 paraffin-embedded IBCs were immunohistochemically stained with a monoclonal antibody to maspin. A nuclear signal was present in 96% and a cytoplasmic signal in 35% of the cases. Nuclear staining was related to oestrogen (ER) and progesterone receptor (PR) positivity (p < 0.0001), but not to S-phase fraction (SPF) or ploidy. Cytoplasmic staining was related to ER and PR negativity (p < 0.0001), high SPF (p < 0.0001), and aneuploidy (p = 0.003). Thus, maspin nuclear staining was significantly associated with good prognostic factors, while cytoplasmic staining was associated with poor prognostic markers. These findings suggest that the presence of maspin in two different compartments of the cell may have different biological and clinical implications. Additional studies are needed to evaluate further this expression profile of maspin in breast cancer. PMID- 12635134 TI - Absence of either gastric or intestinal phenotype in microscopic differentiated gastric carcinomas. AB - Differentiated gastric carcinoma (DGC) corresponds roughly to the intestinal type of gastric carcinoma described by Lauren. It has been suggested that DGCs arise from intestinalized gastric mucosa, but recent findings regarding their mucin expression do not support this hypothesis. To evaluate the histogenetic relationship between DGCs and intestinal metaplasia, lesions that are as small as possible should be examined. Twenty-five DGCs, ranging in their greatest dimension from 0.4 to 2.7 mm, were collected and divided into two groups by size. Group A consisted of 13 lesions less than 1.4 mm across, and group B of 12 lesions 1.4 mm or more. The presence of mucin and a brush border was assessed by immunostaining with antibodies against human gastric mucin, pyloric-gland-type mucin, Muc-2 glycoprotein, and CD10 antigen, and the lesions were classified as having the gastric phenotype (G-type), intestinal phenotype (I-type), mixed gastric and intestinal phenotype (M-type), or null phenotype (N-type). Thirteen (52%) of the 25 lesions were N-type, 5 (20%) lesions were G-type, 5 (20%) were I type, and 2 (8%) were M-type. Group A had a larger proportion of N-type lesions than B (10/13, or 77%, vs. 3/12, or 25%; p = 0.027, chi-square test for proportions). Group B had a larger proportion of G-type lesions than A (5/12, or 42%, vs. 0/13, or 0%; p = 0.033). The phenotypes of the carcinomas and their surrounding mucosa were unrelated. Therefore, DGCs may arise from gastric mucosa affected by intestinal metaplasia or not, without having either the gastric or intestinal phenotype. PMID- 12635135 TI - Epstein-Barr virus and microsatellite instability in gastric carcinogenesis. AB - Little information is available concerning the relationship between transforming viruses and microsatellite instability (MSI). We evaluated Epstein-Barr virus (EBV) using in situ hybridization for EBV-encoded small RNAs and MSI using the polymerase chain reaction in surgically resected gastric cancer. The study subjects included 298 consecutive cases of solitary gastric carcinoma, 63 gastric carcinomas in young patients ( 1 x 10(9) particles/gram. PMID- 12635160 TI - Macrophages at the skeletal tissue-device interface of loosened prosthetic devices express bone-related genes and their products. AB - Aseptic loosening of prosthetic arthroplasty is the most common reason for implant failure in adult orthopaedic reconstruction. At the interface of aseptic loosened prostheses, there is an abundance of particle-activated macrophages and other inflammatory cells. The role of these particle-laden macrophages in the osteogenic arm of the remodeling skeleton in this pathological condition is poorly understood. Molecular signaling by mesenchymal cells and mononuclear inflammatory cells residing in the interfacial tissues between bone and cement or prosthetic material of aseptically loosened joint prostheses was studied using in situ hybridization and immunohistochemical techniques. We found that a range of collagenous and noncollagenous matrix proteins, including osteopontin, osteocalcin, bone sialoprotein, and type I collagen, were produced in the periprosthetic tissue by foamy macrophages, as well as nearby osteogenic cells. The former accumulated in profusion in the three zones of interfacial tissues: pseudomembranous, fibrous, and osseous. Spindle mesenchymal cells in the fibrous zone failed to express any of the osteogenic mRNAs or proteins sought. The expression of bone-related genes and proteins by foamy macrophages at the interface of an aseptic loosened prosthesis may contribute to the disturbance of bone remodeling at this site. PMID- 12635161 TI - Proanthocyanidin: a natural crosslinking reagent for stabilizing collagen matrices. AB - While attempting to find a suitable crosslinking reagent for biopolymers, a naturally occurring proanthocyanidin (PA) obtained from grape seeds was selected to fix biological tissues. The cytotoxicity and crosslinking rate, reflected by the in vitro and in vivo degradation of fixed matrices has been studied. The shrinkage temperature of the fixed bovine pericardium increased from 66 to 86 degrees C. A cytotoxicity assay using fibroblast cultures revealed that PA is approximately 120 times less toxic than glutaraldehyde (GA), a currently used tissue stabilizer. In vitro degradation studies showed that fixed tissue was resistant to digestion by bacterial collagenase. Crosslinks between PA and tissues can be stabilized by decreasing the dielectric constant of the solution during storage. After subcutaneous implantation for periods ranging between 3 and 6 weeks, we found no apparent degradation of the GA- or PA-fixed tissues, whereas fresh tissue controls rapidly disintegrated. Beyond 6 weeks PA crosslinks began to degrade. More fibroblasts migrated and proliferated inside the PA-fixed implants compared with GA counterparts. Tissues crosslinked with PA manifested an enhanced collagen expression and deposition and did not calcify after implantation. GA, on the other hand, even after thorough rinsing continued to be cytotoxic, inhibited collagen synthesis and encouraged dystrophic calcification. Collagen matrices crosslinked with PA are expected to be of value in the design of matrices that will encourage cell ingrowth and proliferation, which are temporary in nature, and that are intended to regenerate or replace missing tissues, which can delay the biogradation of collagen. As such they should be of significant value in the emerging field of tissue engineering. PMID- 12635162 TI - Swinging the vote for rapamycin. PMID- 12635163 TI - Trapping technology. PMID- 12635165 TI - One size fits all. PMID- 12635164 TI - A perfect mismatch. PMID- 12635166 TI - Let's stick together. PMID- 12635167 TI - Drifting downstream. PMID- 12635168 TI - Balancing act. PMID- 12635169 TI - Top scorer. PMID- 12635170 TI - Age activator. PMID- 12635171 TI - Specialization of tumour vasculature. AB - Tumour blood vessels express markers that are not present in resting blood vessels of normal tissues, but that can be shared by angiogenic vessels in non malignant conditions. Many of these proteins are functionally important in the angiogenic process. Some tumours also contain lymphatic vessels, as well as channels that consist of cancer cells and their extracellular matrix. These special features of tumour vessels are good targets for cancer therapies. PMID- 12635172 TI - Role of integrins in cell invasion and migration. AB - As cancer cells undergo metastasis--invasion and migration of a new tissue--they penetrate and attach to the target tissue's basal matrix. This allows the cancer cell to pull itself forward into the tissue. The attachment is mediated by cell surface receptors known as integrins, which bind to components of the extracellular matrix. Integrins are crucial for cell invasion and migration, not only for physically tethering cells to the matrix, but also for sending and receiving molecular signals that regulate these processes. PMID- 12635173 TI - Endocrine-responsive breast cancer and strategies for combating resistance. AB - Deaths from breast cancer have fallen markedly over the past decade due, in part, to the use of endocrine agents that reduce the levels of circulating oestrogens or compete with oestrogen for binding to its receptor. However, many breast tumours either fail to respond or become resistant to endocrine therapies. By understanding the mechanisms that underlie this resistance, we might be able to develop strategies for overcoming or bypassing it. PMID- 12635174 TI - Modifiers of risk of hereditary breast and ovarian cancer. AB - Hereditary breast and ovarian cancer is among the most commonly encountered adult genetic disease, and it is increasingly important that geneticists, oncologists, surgeons and gynaecologists are aware of the issues regarding risk assessment, prevention and management of women with inherited susceptibility to cancer. Genetic risk can be modified by external factors, but what are these factors, and how might our knowledge of them help us to better define the risks for individual women and to develop strategies for cancer prevention? PMID- 12635175 TI - Second cancers in survivors of childhood cancer. AB - More than 70% of children diagnosed with cancer can now be expected to be long term survivors. However, the consequences of 'cure' might be considerable for the survivors of cancer: 60-70% of young adults who have survived childhood cancer will develop at least one medical disability as a result of their cancer or, more commonly, as a result of their therapy. Of these, the most devastating is a second cancer. PMID- 12635176 TI - RHO-GTPases and cancer. AB - The RAS oncogenes were identified almost 20 years ago. Since then, we have learnt that they are members of a large family of small GTPases that bind GTP and hydrolyse it to GDP. This is then exchanged for GTP and the cycle is repeated. The switching between these two states regulates a wide range of cellular processes. A branch of the RAS family--the RHO proteins--is also involved in cancer, but what is the role of these proteins and would they make good therapeutic targets? PMID- 12635177 TI - Natural products in cancer chemotherapy: past, present and future. AB - Natural products have been the mainstay of cancer chemotherapy for the past 30 years. However, the quickening pace of (aberrant) gene identification, and the new technologies of combinatorial chemistry and high-throughput screening, should provide access to a wide range of new, totally synthetic drugs. Will these new approaches sound the death knell for therapies based on natural products? In reality, natural products are likely to provide many of the lead structures, and these will be used as templates for the construction of novel compounds with enhanced biological properties. PMID- 12635178 TI - Hope at last for European collaboration. PMID- 12635180 TI - Cancer research. Detrick unveiled. PMID- 12635179 TI - Technology and the new federalism. PMID- 12635181 TI - Links of iron. PMID- 12635183 TI - Strand switches in SV40 transcription. PMID- 12635182 TI - Prostaglandins. Promise but no miracles. PMID- 12635184 TI - Differential recognition immune responses. PMID- 12635185 TI - Aggregation factors and receptors. PMID- 12635186 TI - Environmental health. Moderation is the word. PMID- 12635187 TI - Oncolytic viruses. PMID- 12635188 TI - Messenger RNA. The missing poly A tail. PMID- 12635189 TI - Ionosphere. Heat probing at Arecibo. PMID- 12635190 TI - Why Venus is cloudy. PMID- 12635191 TI - Value of objective examinations. AB - In some cases the traditional essay examination question is being superseded by more objective tests, requiring straightforward answers. If administered carefully these can provide a reliable means of assessment, but much research remains to be done. PMID- 12635192 TI - Measurement from Gauss to Whitehead. PMID- 12635193 TI - Nature of haem-haem interaction. AB - Haem-haem interaction consists of a change of tension at the haems, caused by a transition between two alternative quaternary structures of the protein. Dr Perutz describes how spin changes that accompany reaction with ligands alter the oxygen affinity of the haems. PMID- 12635195 TI - Evidence for solar particle production above approximately 75 GeV. AB - Observations of solar particles in the energy region above approximately 75 GeV provide good evidence for particle acceleration during the initial phase of a solar flare. PMID- 12635194 TI - Synthetic RNA-dependent DNA polymerase activity in normal rat liver and hepatomas. AB - Normal adult rat liver contains a high level of a synthetic RNA-dependent DNA polymerase activity, which is distinct from cellular DNA-dependent polymerases. It uses poly(dT).poly(rA) and poly(rA).poly(rU) as templates but has little or no response to DNA or several single-stranded RNAs. PMID- 12635196 TI - Radio detection of Cygnus X-3. PMID- 12635197 TI - Radio observations of Cygnus X-3. PMID- 12635198 TI - LR Cen is not Cen X-3. PMID- 12635199 TI - Ring current effect on magnetospheric electron density profiles derived from plasmapause whistlers. PMID- 12635200 TI - Super-resolution aperture scanning microscope. PMID- 12635201 TI - Liquid phase synthesis of peptides. PMID- 12635202 TI - Ouabain and the onset of the blood-brain barrier in neonate chicks. PMID- 12635203 TI - Experimental infection of chimpanzees with the virus of hepatitis B. PMID- 12635204 TI - Bioluminescence in the marine teleost, Porichthys notatus, and its induction in a non-luminous form by Cypridina (ostracod) luciferin. PMID- 12635205 TI - Transfer of inorganic mercury to milk of goats. PMID- 12635206 TI - Enhancement of bacterial methane oxidation by clay minerals. PMID- 12635207 TI - Reversed geomagnetic events in the Brunhes epoch. PMID- 12635208 TI - LASA's window cleaned? PMID- 12635209 TI - Fracture of polymers. PMID- 12635210 TI - Aseismic ridge dredge hauls. PMID- 12635211 TI - Replicating DNA. PMID- 12635212 TI - Cyclic AMP, cell surfaces and mitosis. PMID- 12635213 TI - Acclimatized rats. PMID- 12635214 TI - More uses for neutrons. PMID- 12635216 TI - Apollo clock corrections. PMID- 12635215 TI - Inside cool interstellar clouds. PMID- 12635217 TI - Two hundred years of the Radcliffe Observatory. AB - Dr Thackeray, the Radcliffe Observer, traces the history of the Radcliffe Observatory at Oxford and later in South Africa. PMID- 12635218 TI - Error theory and ageing in human diploid fibroblasts. AB - Immunological and enzymatic assessments of lactate dehydrogenase in human lung fibroblasts strongly suggest that altered proteins accumulate in ageing cells. This supports but does not prove the error catastrophe theory of all death. PMID- 12635219 TI - Lateral heterogeneity in the Earth's mantle. AB - The characteristics of seismic waves detected at the Large Aperture Seismic Array in Montana suggest that, beneath the Bonin Island Arc in particular, there are lateral inhomogeneities in the mantle. PMID- 12635221 TI - Her X-1: a precessing binary pulsar? PMID- 12635220 TI - Radio continuum emission at 21 cm near Stephan's Quintet. PMID- 12635222 TI - A sediment cloud below the Mediterranean outflow. PMID- 12635223 TI - Evidence of a geomagnetic excursion 30,000 yr BP. PMID- 12635224 TI - Subduction zones: not relevant to present-day problems of waste disposal. PMID- 12635225 TI - Crystal structure of a novel curariform agent. PMID- 12635226 TI - Growth-promoting effects of silicon in rats. PMID- 12635228 TI - Opposite effects of L-dopa and 5-HTP on spinal sympathetic reflexes. PMID- 12635227 TI - Blood pressure decreased by halobutylpicolinic acid, an inhibitor of dopamine beta-hydroxylase. PMID- 12635229 TI - Heart rate-cloacal temperature hysteresis in Iguana is a result of thermal lag. PMID- 12635230 TI - Pre-courtship activity and a new scent organ in butterflies. PMID- 12635231 TI - Thyroid of the coelacanth, Latimeria chalumnae Smith. PMID- 12635232 TI - Visual pigment of the coelacanth. PMID- 12635233 TI - Ion permeability of phospholipid bilayers. PMID- 12635235 TI - Grass evolution, the Kranz syndrome, 13C/12C ratios, and continental drift. PMID- 12635234 TI - Photoconduction in proteins. PMID- 12635236 TI - Terrorism and Vietnam. PMID- 12635238 TI - Indian science. PMID- 12635237 TI - Doomwatching. PMID- 12635239 TI - Pyrrhic victories for the friends of whales. PMID- 12635240 TI - Plant breeding. Genes for all seasons. PMID- 12635241 TI - Second nature? PMID- 12635242 TI - World health organization. Genetic diseases. PMID- 12635243 TI - Channel tunnel. Decision by degrees. PMID- 12635244 TI - Jostling for space in Orbit. PMID- 12635245 TI - Mistakes in ageing fibroblasts. PMID- 12635246 TI - Intensity of solar neutrinos. PMID- 12635247 TI - Strand switching during SV40 transcription. PMID- 12635248 TI - Ribosomes. Protein gene expression. PMID- 12635249 TI - Artificial intelligence. Understanding program. PMID- 12635250 TI - A structure for thermolysin. PMID- 12635251 TI - Hard X-ray flare in cygnus X-1. PMID- 12635252 TI - Bioengineering. Catch-all or cornucopia? PMID- 12635253 TI - Oceanic fishes. Biology of bramids. PMID- 12635254 TI - Messenger RNA inactivation. PMID- 12635255 TI - DNA structure. Geometrical versatility. PMID- 12635257 TI - Abortive transcription as reovirus matures. PMID- 12635256 TI - Transcription and excision of prophage lambda. PMID- 12635258 TI - Antibody production and skin graft rejection. PMID- 12635259 TI - Parity and optical activity. AB - Simple pictorial arguments can demonstrate that the natural and Faraday rotation experiments conserve parity and time-reversal invariance. The method can be used to predict possible new effects without recourse to mathematical theories. PMID- 12635260 TI - Can the lumpy distribution of galaxies be detected by X-ray observations? AB - Published data from X-ray telescopes are not adequate to reveal the fluctuations in the surface brightness of the background radiation caused by clustering or superclustering of the sources giving rise to the background. PMID- 12635261 TI - What cooks with solar neutrinos? AB - Two desperate explanations of the solar neutrino puzzle are proposed, one involving experimental nuclear physics and the other theoretical solar structure and evolution. PMID- 12635262 TI - Altered enzymes in ageing human fibroblasts. AB - Diploid human fibroblasts accumulate heat labile enzymes during the final stages of their life-span in culture. The RNA base analogue 5-FU induces premature senescence, which is preceded by the appearance of altered enzyme. These observations support Orgel's "error catastrophe" theory of ageing. PMID- 12635263 TI - Galactic gamma-radiation between 200 MeV and 10 GeV. PMID- 12635264 TI - Observations of D-region modifications at low and very low frequencies. PMID- 12635265 TI - Late Cambrian fossils and the age of the Ross Orogeny, Antarctica. PMID- 12635266 TI - The direct viewing and brief-time recording of crystallographic events. PMID- 12635267 TI - Perfluoropolyether--a vacuum pump fluid resistant to electron induced polymerization. PMID- 12635268 TI - Electrochemical photolysis of water at a semiconductor electrode. PMID- 12635269 TI - One and two-dimensional structure of alpha-helix and beta-sheet forms of poly(l alanine) shown by specific heat measurements at low temperatures (1.5-20 K). PMID- 12635270 TI - Performance of composite foetal hearts. PMID- 12635271 TI - Hypothesis for a neuronal mechanism involved in memory. PMID- 12635272 TI - Alpha rhythms and hyperkinesis. PMID- 12635273 TI - Alpha rhythms in the hyperkinetic child. PMID- 12635274 TI - Visual system's view of acoustic space. PMID- 12635275 TI - Improved relationship between the deposition of strontium-90 and the contamination of milk in the United Kingdom. PMID- 12635276 TI - Resistance to synthetic juvenile hormone in a strain of the flour beetle, Tribolium castaneum. PMID- 12635277 TI - Statocyst lacking cilia in the coelenterate Corymorpha palma. PMID- 12635278 TI - Cognitive contours. PMID- 12635279 TI - Terrorism. PMID- 12635280 TI - Science and ethics. PMID- 12635281 TI - Defence and technology in Europe and elsewhere. PMID- 12635282 TI - Evolution of elephants and suids in east Africa. PMID- 12635283 TI - Hydra as a model organism. PMID- 12635284 TI - Difficulties in telling QSOs from blue stars. PMID- 12635285 TI - Lower mantle inhomogeneity. PMID- 12635286 TI - Controlled fusion. Tokamaks to win? PMID- 12635287 TI - Methylation of bovine rhodopsin. PMID- 12635288 TI - Clustering of concanavalin A sites after proteolysis. PMID- 12635289 TI - Proteins. Concanavalin observed. PMID- 12635290 TI - Forty-five varieties of research (and development). AB - Lord Rothschild presents his personal views about the terminology used to describe research and development. PMID- 12635291 TI - Vertebrate faunas and chronology of hominid-bearing sediments east of Lake Rudolf, Kenya. AB - The vertebrate faunas which have been collected from east of Lake Rudolf include hominid remains and the earliest dated stone artefacts known. It is now possible to recognize faunal units and to establish a tentative chronology for observed faunal events. It is suggested here that the faunal sequence at East Rudolf may extend from 4.5 m.y. to about 1.3 m.y. with a significant break between about 4.0 and 2.6 m.y. PMID- 12635292 TI - Immunogenetic model. AB - Different and equally legitimate modes of interpreting serologic reactions will considerably change the image of reality. Some principles of such transformations are briefly outlined and applied to immunogenetic systems. PMID- 12635293 TI - Black holes and temporal ordering. PMID- 12635294 TI - Is the African plate stationary? PMID- 12635295 TI - Glaciation in the Somerset levels: the evidence of the burtle beds. PMID- 12635296 TI - Distribution of cadmium in north Atlantic deep-sea sediments. PMID- 12635297 TI - Cathodoluminescence in deformed diamond. PMID- 12635298 TI - Laser analogue to ball lightning. PMID- 12635299 TI - Antimatter and ball lightning. PMID- 12635300 TI - Enzymatic cleavage of methotrexate provides a method for prevention of drug toxicity. PMID- 12635301 TI - Methadone induced mortality as a function of the circadian cycle. PMID- 12635302 TI - Segregation and sedimentation of red blood cells in ultrasonic standing waves. PMID- 12635303 TI - Stimulation of the sodium pump in the red blood cell by lithium and potassium. PMID- 12635304 TI - Evidence of cross-resistance to a juvenile hormone analogue in some insecticide resistant houseflies. PMID- 12635305 TI - Control of de novo isocitrate lyase synthesis in Chlorella. PMID- 12635306 TI - Quinacrine-like fluorescence of extracts from papaveraceae and fumariaceae. PMID- 12635307 TI - Food-sharing behaviour within a group of douc monkeys (Pygathrix nemaeus nemaeus). PMID- 12635308 TI - Effect of humming on watching television. PMID- 12635309 TI - The Fourier transform of evoked responses. PMID- 12635310 TI - Kammerer's midwife. PMID- 12635311 TI - Faecal pollution of our beaches--how serious is the situation? PMID- 12635312 TI - Disagreements on why brown pelican eggs are thin. PMID- 12635314 TI - Siding Spring. PMID- 12635313 TI - BHC not found to reduce the shell quality of hens' eggs. PMID- 12635315 TI - A counter-homily. PMID- 12635316 TI - [Surgical treatment for esophageal cancer in elderly patients over 75 years of age]. AB - We performed 127 esophageal resections for the esophageal cancer patient from December 1995 to September 2001. It was separated to under 70 years old patients group (group I), 71-74 years old patients group (group II), and over 75 years old patients group (group III). RESULTS: Postoperative complication was occurred in 53 cases (41.7%) within all of 127 esophageal resected cases. It was 33.7% in group I, 53.6% in group II, 62.5% in group III. Four years survival rate of each group is 38.3% in group I, 44.6% in group II, 31.3% in group III. It is significantly better in group II rather than in group III. Operative death rate is 12.5% (2 cases) in group III, 7.1% (2 cases) in group II, 3.6% (3 cases) in group I, and it is gradually higher and higher by patient's age. CONCLUSIONS: (1) In the esophageal cancer patient over 75 years old, postoperative complication rate is higher than under 74 years old patients, and prognosis is significantly poor rather than in 70-74 years old patients group. (2) In the esophageal cancer patient over 75 years old, we considered it is good indication of esophagectomy for stage I and stage II patient without preoperative complication, however, there are no operative indication for stage III and stage IV patient. PMID- 12635317 TI - [Aortic root replacement using cryopreserved allograft for an infected aorta patient associated with mediastinitis]. AB - We herein report a case of aortic root replacement using cryopreserved allograft. A 52-year-old man received aortic valve replacement using a mechanical prosthesis for aortic stenosis. He was complicated by postoperative methicillin-resistant staphylococcus aureus mediastinitis involving the ascending aorta. Surgical therapy including debridement, omental transposition, patch closure using pericardium had not achieved satisfied result. Aortic root replacement using cryopreserved allograft was mandatory for refractory aortic infection. He had an uneventful postoperative course. The cryopreserved allograft was effective for a patient with refractory aortic root infection due to postoperative mediastinitis. PMID- 12635318 TI - [A successful treatment for myonephropathic metabolic syndrome after operation of acute aortic dissection (Stanford type A) with multiple organ ischemia]. AB - A 60-year-old man had sudden chest pain and right lower extremity pain. A diagnosis of acute aortic dissection (Stanford type A) with right lower extremity ischemia was made. An emergency ascending aortic and total arch replacement, right axillo-femoral bypass, and coronary artery bypass grafting was performed for acute aortic dissection, acute right iliac arterial occlusion, and acute myocardial infarction. Myonephropathic metabolic syndrome (MNMS) occurred 2 days after operation. The serum creatine kinase value increased over 68,000 U/l, hemodiafiltration was started on the 2nd postoperative day (POD). The patient fell into anuric state and hemodiafiltration was performed 3 times a week. Urination was obtained over 1,000 ml/day from the 25th POD and hemodiafiltration was not necessary any more. We emphasize that prompt operation including revascularization of ischemic leg should be performed in acute aortic dissection with extremity ischemia and also prompt hemodiafiltration to improve the prognosis of these disastrous lesion. PMID- 12635319 TI - [Preoperative magnetic resonance imaging evaluation of the aortic annular diameter in aortic valve stenosis]. AB - Accurate preoperative knowledge of the aortic annular diameter is a great value in determining the appropriate procedure for aortic valve stenosis. We have successfully used magnetic resonance imaging (MRI) to evaluate the size of the aortic annular diameter preoperatively. Between October 1997 and October 1998, 5 aortic valve replacements were performed in patients with aortic valve stenosis. MRI was performed preoperatively in all cases. All images were gated to the R wave of the electrocardiogram. Images representing 5 mm slices were acquired in the plane which is parallel to the line through the center points of the aortic root and left ventricle outflow tract. The maximum diameter of the aortic annulus (D) was measured at the point just below the attachment of the aortic valve leaflets. This diameter was compared to the maximum diameter of the prosthetic valve sizer (D 1) which could be easily inserted into the aortic annulus intraoperatively, and to the diameter of the prosthetic valve (D 2) that ultimately was implanted. Twenty-five minutes were required to scan each patient. There were no complications related to scanning. Mean values were D = 23.4 mm, D 1 = 22.3 mm and D 2 = 21.4 mm. There was a significant correlation between D and D 1, and D 2 was approximately 2 mm smaller than D. MRI evaluation is an accurate, noninvasive method for determining the aortic annular diameter preoperatively. We highly recommend this method for measuring the aortic annular diameter in case of aortic valve stenosis as part of the preoperative evaluation. PMID- 12635320 TI - [Analysis of continuous hemodiafiltration after open heart surgery]. AB - The development of renal failure after open heart surgery is associated with a high mortality. Thirteen patients were treated with continuous hemodiafiltration (CHDF) for renal failure following open heart surgery since April 1999 to December 2001. The indication of CHDF was blood purification in 8 patients and water balance control in 5 patients. Two patients with hemodialysis (HD) before operation returned to usual HD early after operation, and discharged. One patient died of severe heart failure, and another patient died of sepsis and multi organ failure. In these 2 patients, CHDF could not withdrawn. Seven patients weaned from CHDF 1 to 19 days after operation. Five of 7 patients discharged, but 2 patients died of cerebral infarction 4 month after operation. The remaining 2 patients could not wean from CHDF, and were introduced HD. Only 1 of 13 patients had bleeding tendency (cardiac tamponade). CHDF did not influence the hemodynamic state and was very effective for the treatment of renal failure in many patients. PMID- 12635321 TI - [Radiofrequency ablation for metastatic lung tumor]. AB - BACKGROUND: Radiofrequency ablation (RFA) is a thermal ablation technique which results in coagulative necrosis. Several preliminary studies have evaluated the efficacy of RFA for liver tumor and benign prostate disease, but few studies for lung tumor. We experienced RFA for a metastatic lung tumor and it was effective. CASE: A 61-year-old woman was admitted to our hospital with chest pain. Extended thymothymomectomy with lymph node dissection was performed after diagnosis of thymic carcinoid. After 14 months, multiple small lung nodules, which had been 5 10 mm in diameter, were appeared and resected. However, 23 months after the first operation and 14 months after the second operation, multiple small lung nodules were appeared again and RFA was performed for 1 tumor without any complications. We used RITA 500 PA (RITA Medical Systems, INC.) and quadruple-hook radiofrequency probe with electrodes. Under local anaesthesia and ultrasound guidance, the probe was inserted into the target tumor and the hooks were deployed. The tumor became smaller (35 mm to 25 mm in diameter) in 2 months. CONCLUSION: RFA is an effective and minimally invasive technique to destruct local tumors. PMID- 12635322 TI - [Hemolytic anemia due to aortic valve regurgitation after mitral valve replacement]. AB - A 50-year-old woman was admitted to our hospital because of heart failure (NYHA III) due to mitral valve regurgitation (MR) with pulmonary hypertension (PH) and tricuspid valve regurgitation (TR). She had a history of chronic renal failure undergoing dialysis (peritoneal dialysis, homodialysis) since 1996. Cardiac catheterization and ultrasonic cardiography showed severe MR (Sellers III), severe TR and PH (mean pressure 33 mmHg). So we performed mitral valve replacement and tricuspid annuloplasty (DeVega). Frequent blood transfusion was needed because severe hemolytic anemia appeared after operation. Ultrasonic cardiography demonstrated moderate aortic valve regurgitation (AR) with no paravalvular prosthetic leakage. We diagnosed hemolytic anemia due to AR. We performed aortic valve replacement. Hemolytic anemia improved soon after second operation. We investigated the mechanical process of the AR. She had a very short subaortic curtain (5.9 mm) compared with the average (8.7 +/- 2.1 mm: mean +/- SD) of cardiac patients. We think that we must be very careful with suture to short subaortic curtain. In addition measurement of subaortic curtain before operation is very useful. PMID- 12635323 TI - [Complete disruption of cervical trachea, whose distal airways were migrated into the mediastinum]. AB - The first case was a 55-year-old man, who suffered by a rope while driving his motor bicycle. On 7th day after injury, tracheotomy was scheduled due to progressive dyspnea. Following intubation of a endotracheal tube, his trachea was ruptured. The second case was a 16-year-old man, who was stabbed his trachea with a sword by his mother. His trachea completely separated following coughing during the examination of bronchoscopy. For 2 cases, we immediately excised their necks for tracheotomy but couldn't find their distal portion of trachea, because they were migrated into the mediastinum. We inserted our finger into the mediastinum for exploration and could draw it back. Both case's postoperative course was uneventful. Whenever cervical trachea is completely separated, tracheal distal end may be pulled down into the mediastinum. We invited new technique of exploration for migrated trachea using our finger. PMID- 12635325 TI - [Removal of methicillin-resistant staphylococcus aureus infected pacemaker leads under cardiopulmonary bypass: report of a case]. AB - A 71-year-old man, who underwent an intravenous pacemaker implantation previously, suffered from fever and local infection of the generator pocket. A blood culture showed positive for methicillin-resistant staphylococcus aureus (MRSA). He underwent removal of total pacemaker system under cardiopulmonary bypass support successfully. Two leads were tightly adhered to the right atrial free wall, tricuspid valve and right ventricular trabeculation. Postoperative course was uneventful with administration of antibiotics for 5 weeks. Removal under cardiopulmonary bypass is considered to be an effective procedure for treatment of patients with infected pacemaker lead. PMID- 12635324 TI - [Stent-graft implantation for the patient with ruptured acute aortic syndrome: report of a case]. AB - We report a ruptured case of acute aortic syndrome and a successful treatment with transcatheter placed stent-graft. A 67-year-old man underwent a stent-graft implantation for the patient with ruptured acute aortic syndrome. The patient admitted in our institute for a diagnosis of ruptured acute aortic dissection. We planned to operate a graft replacement for distal arch aorta. But he was complicated with right-side monoplesia before the operation. Then we performed a stent-graft implantation in order to avoid the usage of extra-corporeal circulation. The implantation of the stent-graft was successful and shrinkage of the hematoma around the ruptured aorta was documented on the computed tomography of 7 post-operative day. PMID- 12635326 TI - [Surgical treatment for giant ascending aorta-arch aneurysm coexisted with DeBakey type II dissection on elderly women: report of a case]. AB - A 78-year-old woman was admitted to our hospital after computed tomography (CT) had revealed in her the presence of a giant ascending aorta-arch aneurysm. This aneurysm was about 8 cm in diameter and associated with DeBakey type II dissection. Aortography showed the same condition as the CT view with the entry on the ascending aorta. The ascending aortaarch was replaced with a Hemashield 24 mm, by using deep-hypothermic selective cerebral perfusion and the open distal method. There were no complications during her peripostoperative state and no evidence of leakage and remnant dissection on CT and aortography. This is a rare case in which thoracic aortic aneurysm coexisted with dissection. In this case of severe atherosclerosis, deep-hypothermic selective cerebral perfusion and the open distal method provided effective treatment. PMID- 12635327 TI - [A giant left atrial myxoma with severe mitral valve regurgitation: report of a case]. AB - A 76-year-old female underwent operation with a diagnosis of a left atrial myxoma with accompanied mitral regurgitation. Although no clinical findings of mitral regurgitation were noticed preoperatively, degenerative changes to the anterior leaflet as well as chordae tendinae possibly due to mechanical damage by the movement of the giant tumor through the mitral valve complex were observed in operation. Resection of the tumor and mitral valve replacement were successfully performed. Our case suggests that it is indispensable to investigate the mitral valve during operation even in case of the left atrial tumor with no preoperative findings of mitral regurgitation. PMID- 12635328 TI - [Aortic valve repair with minimal access "J" sternotomy]. AB - Recently many cardiovascular operations have been performed with minimally invasive cardiac surgery (MICS). It is reported that MICS is superior to standard full sternotomy in regard to cosmetic and post-operative recovery. In our institution, aortic valve repair operations have been performed to aortic valve insufficiency. This time, we performed aortic valve repair by minimal access "J" sternotomy. A 63-year-old male with degenerative aortic regurgitation underwent aortic valve repair by MICS. He had experienced dyspnea on exertion 7 days before admission. Preoperative transthoracic echocardiogram showed the grade of aortic regurgitation (AR) was severe. Surgery was successful and the grade of AR became trivial. This combined procedure have merits for patients because of freedom from anticoagulation and small incision. Such combined surgery may be indicated in the treatment in a subset of patients with degenerative AR. PMID- 12635329 TI - [Bilateral coronary ostial stenoses with aortic regurgitation in a patient with syphilitic aortitis]. AB - Syphilitic aortitis is now rare in developed countries and is sometimes overlooked. A 61-year-old man with bilateral coronary ostial stenoses (#5:90%, #1:99%) and Sellers III/IV aortic regugitatioin (AR) induced by syphilitic aortitis presented with chest pain. Preoperative rapid plasma reagin titer and Treponema pallidum hemagglutination test were strongly positive, 256 fold and 191.25 C.O.I., respectively. Aortic valve replacement (AVR) and coronary artery bypass grafting (CABG) with bilateral internal thoracic arteries (ITA) was performed successfully. The angiographic features as follows: 1) coronary artery stenosis is generally limited to the ostia, 2) the grade of stenosis is almost always more than 90%, 3) AR is frequently associated with coronary ostial stenosis. CABG should be performed with ITA, not saphenous vein grafts, to avoid occlusion of the ostium of the saphenous vein graft by syphilitic aortitis. Retrograde cardioplegia should be performed if ostial stenosis is severe. PMID- 12635330 TI - [Coronary artery aneurysm developed acute myocardial infarction: report of a case]. AB - A 68-year-old woman without angina developed acute antero-septal myocardial infarction. Coronary angiogram revealed saccular coronary artery aneurysm at the origin of the diagonal branch and relatively mild coronary artery stenosis. Emboli from the coronary aneurysm was strongly suggested. We placed coronary bypass grafts onto left anterior descending, diagonal and obtuse marginal branches, and excluded the coronary aneurysm by ligating both its inflow and outflow. The postoperative course was uneventful. There are several treatments of choice for patients with coronary aneurysm. We believe that surgery is mandatory and that exclusion of the coronary aneurysm should be considered in every possible case. PMID- 12635331 TI - [Pulmonary actinomycosis mimicking of mucosa-associated lymphoid tissue lymphoma by frozen section examination: report of a case]. AB - A 59-year-old man was admitted to our hospital for hemoptysis. Chest X-ray films showed a mass shadow in the left lower lobe. Any definitive diagnosis could not be made after examinations including sputum culture and bronchial cytology. Exploratory thoracotomy was performed since malignancy was not excluded. During the operation, mucosa-associated lymphoid tissue (MALT) lymphoma was suspected based on the pathological findings of the frozen section. The permanent section of the resected specimen contained colonies of actinomycetes in the bronchus, and the peripheral region demonstrated marked infiltration of lymphocytes and plasma cells through the interstitium of alveolar walls and peri-bronchovascular sheath, with many lymphoid follicles. This case revealed pulmonary actinomycosis coexisted with lymphocytic interstitial pneumonia (LIP). We concluded that lymphoproliferative disease should be considered another candidate for the differential diagnosis from pulmonary actinomycosis. PMID- 12635333 TI - [Inhibition of bacterial growth in thawed cake and rapid evaluation methods for microbiological quality]. AB - The inhibition of bacterial growth in thawed cake kept under refrigeration and rapid evaluation methods for microbiological quality of the cake were investigated. The effects of the freezing temperature and the addition of ethanol or emulsifier on bacterial numbers in a cake model after storage for 72 hr at 10 degrees C following the thawing process were also studied. Bacterial growth in the cake model was inhibited by the additives under various freezing conditions. In addition, rapid evaluation methods for estimating bacterial numbers in the cake model after incubation for 72 hr at 10 degrees C were studied. High correlations were found between bacterial numbers in the cake model incubated for 24 hr at 20 degrees C and for 6 hr at 35 degrees C with tryptic soy broth and that of the cake model incubated for 72 hr at 10 degrees C. This result indicated that rapid evaluation by incubation for 24 hr at 20 degrees C or for 6 hr at 35 degrees C with tryptic soy broth can be used to predict the bacterial numbers in a cake model after incubation for 72 hr at 10 degrees C. Furthermore, the ATP bioluminescence method was applied to shorten the testing time, because culture on an agar medium was not necessary. PMID- 12635332 TI - [Penetrating cardiac and pulmonary injury by a nail-gun: report of a case]. AB - A 64-year-old man underwent emergency operation for self-inflicted penetrating cardiac and pulmonary injury by a nail-gun. Chest films on admission showed 5 nails penetrating the thorax and one of these nails had reached into pericardium. Emergency surgery was performed by median sternotomy and the 5 nails in the chest wall were totally removed and the wound of right ventricle and lung were closed with direct mattress sutures without cardio-pulmonary bypass. Post operative course was uneventful and he was transferred to the psychiatric ward for the treatment of depression at post operative 7 days. PMID- 12635334 TI - [Simultaneous determination of N-methylcarbamates and their metabolites in citrus fruits by HPLC avoiding use of halogenic solvents]. AB - Twenty-one N-methylcarbamates (NMCs) and 12 of their metabolites or isomers in citrus fruits were simultaneously determined avoiding use of dichloromethane. NMCs in lemon, orange, and grapefruit were extracted with acetone, then the acetone was evaporated off and sodium chloride was added before extraction with ethyl acetate. The extract was evaporated and the residue was cleaned up on a combined mini-column set of Supelclean ENVI-Carb and Mega Bond Elut SAX cartridges. NMCs were determined by HPLC with post-column reaction and fluorescence detection. All of the NMCs in the orange sample were determined without interfering peaks. However 8 NMCs in lemon extract and 10 NMCs in grapefruit extract were not detected because interfering peaks appeared at similar retention times to those of the NMCs. These NMCs were determined using LC/MS (SIM) and were well recovered. Eighty-three data sets obtained by HPLC and LC/MS showed good similarity, with r2 = 0.9178. Recoveries were 60.1 to 97.8% for major NMCs at a fortification level of 0.1 ppm. The limit of detection by HPLC was 0.005 ppm NMCs in samples and a similar level applied to LC/MS. PMID- 12635335 TI - [Comparison of soyasaponin and isoflavone contents between genetically modified (GM) and non-GM soybeans]. AB - Soyasaponins and isoflavones are main secondary metabolites in soybeans. In this report we compared the content of secondary metabolites between genetically modified (GM) and non-GM soybeans. Six cultivars/lines of GM and six cultivars/lines of non-GM soybeans were extracted with methanol. Each extract was partitioned with aqueous methanol and hexane and the aqueous methanol fraction was partially purified by HP-20 and LH-20 column chromatography to afford crude soyasaponin and isoflavone fractions. The main A-type soyasaponin, acetylsoyasaponin A1 (AcA1), and the main B-type soyasaponins, soyasaponins I and II (I and II), in the crude fractions were identified by LC/MS analyses with authentic samples. The main isoflavones, daidzin, genistin, daidzein and genistein (DI, GI, DE and GE), in the crude fractions were identified by LC photo diode array analyses with authentic samples. The contents of AcA1, I and II in the crude soyasaponin fractions and those of DI, GI, DE and GE in the crude isoflavone fractions were analyzed by reversed-phase HPLC. The average contents (mg/100 g) of AcA1, I, II and total of B-type soyasaponins in GM soybeans were 36.4 +/- 24.2, 51.2 +/- 11.8, 26.4 +/- 7.6 and 77.7 +/- 18.5, respectively, and those in non-GM ones were 22.3 +/- 14.7, 46.3 +/- 17.8, 19.8 +/- 9.1 and 65.9 +/- 26.9, respectively. The average contents (mg/100 g) of DI, GI, DE, GE and total isoflavones in GM soybeans were 93.1 +/- 15.5, 121.8 +/- 19.4, 0.073 +/- 0.178, 0.320 +/- 0.082 and 215.3 +/- 33.3, respectively, and those in non-GM ones were 78.8 +/- 34.6, 106.7 +/- 28.3, 2.206 +/- 4.468, 0.822 +/- 0.754 and 188.5 +/- 26.7, respectively. There were no statistically significant differences in soyasaponin and isoflavone contents between GM and non-GM soybeans. Therefore, it was estimated that the GM soybeans are equivalent to the non-GM ones in terms of the composition of the main secondary metabolites. PMID- 12635336 TI - Specific determination of bromate in bread by ion chromatography with ICP-MS. AB - A sensitive method for detecting bromate in bread by ion chromatography with inductively-coupled plasma mass spectrometry (IC/ICP-MS) was developed. Bromate was extracted from bread with water. The clean-up procedure included a 0.2 micron filter, a C18 cartridge for defatting, a silver cartridge to remove halogen anions, a centrifugal ultrafiltration unit to remove proteins, and a cation exchange cartridge to remove silver ions. A 500 microL sample solution was applied to IC/ICP-MS. The detection limit and the quantitation limit of bromate in the solution were 0.3 ng/mL and 1.0 ng/mL, expressed as HBrO3, respectively, which corresponded to 2 ng/g and 5 ng/g, respectively, in bread. Recovery of bromate was about 90%, and the CV was about 2%. Based on the detection limit in solution and recovery from bread, the detection limit of bromate in bread was estimated to be 2 ng/g. PMID- 12635337 TI - [Constituent of natural food additive hokosshi extract and an analytical method for the additive in foods]. AB - Hokosshi extract is obtained by ethanol extraction from the seeds of hokosshi (Psoralea corylifolia), which is used as a Chinese medicine. The constituents of hokosshi extract were analyzed. The main constituent was isolated using column chromatography, and identified as bakuchiol by TLC, LC/MS and NMR. Bavachinin A was also detected. In order to prepare a marker substance for hokosshi extract, bakuchiol was isolated from seeds of hokosshi using Sep-Pak cartridges. An analytical method for hokosshi extract in foods based on detection of bakuchiol was developed. Bakuchiol was extracted from food with 60 vol% ethanol. The extract was cleaned up using a Sep-Pak plus C18 cartridge, and bakuchiol was determined by HPLC. Seasoning and juice were spiked with hokosshi extract at 500 micrograms/g and analyzed by the proposed method. The recoveries of bakuchiol were 72-99%. The detection limit for the assay was 25 micrograms/g. PMID- 12635338 TI - [Simple analysis of maleic hydrazide in agricultural products by HPLC]. AB - A simplified HPLC determination method for maleic hydrazide in agricultural products was developed, and commercial agricultural crops were investigated. The homogenate of agricultural products was extracted with water. The crude extract was purified on an ACCUCAT Bond Elut extraction cartridge using water. Maleic hydrazide was analyzed by HPLC with UV detection (303 nm). The HPLC separation was performed on a ZORBAX SB-Aq column with acetonitrile-water-phosphoric acid(5:95:0.01) as the mobile phase. Recoveries of maleic hydrazide from 15 agricultural products fortified at 1.0 and 10 micrograms/g were in the ranges of 92.6-104.9% and 94.2-101.3%, respectively. The limit of detection was 0.5 microgram/g in samples. The proposed method was applied to the determination of 242 commercial vegetables and fruits. Maleic hydrazide was detected in 2 samples of imported onion at the levels of 4.9 and 7.2 micrograms/g. PMID- 12635339 TI - [Identification of chemical detected in kiwi fruit during analysis for residual pesticide]. AB - An unknown peak was detected in a GC chromatogram of many kiwi fruit extracts during analysis for pesticide residues. It was identified by GC/MS as diphenyl 2 ethylhexyl phosphate (DPEHP), used as a plasticizer and flame retardant. The concentration of DPEHP was investigated in 15 samples of kiwi fruit, and it was detected at between 0.02 and 0.14 microgram/g in 10 of the samples. It might be due to migration of DPEHP into the fruit from the printed portion of the polyethylene terephthalate (PET) package. PMID- 12635340 TI - [Performance study of analytical method for ethoxyquin in fruit]. AB - A performance study of an analytical method of ethoxyquin (ET) in apples and pears was conducted. In the proposed method, the sample was homogenized with thiourea and ET was extracted with acetone instead of hexane used in the official analytical method for ET, in order to improve the extraction efficiency. Furthermore, dibutyl hydroxytoluene was added in the test solution to prevent the decomposition of ET. For evaluation of the method. ET spiked into apples and pears at the level of 0.2 microgram/g was determined in replicate in six laboratories. Mean recoveries from apple, pear and Japanese pear were 85.3, 83.0 and 83.9%, respectively. Repeatability relative standard deviation values were 3.8-4.7% and reproducibility relative standard deviation values were 7.8-11.3%. The detection limits were 0.001-0.025 microgram/g in the six laboratories; this value may reflect instrument performance. ET spiked into apples and pears at the level of 0.2 microgram/g was detected by both LC/MS and GC/MS. PMID- 12635341 TI - Determination of acrylamide in foods by GC/MS using 13C-labeled acrylamide as an internal standard. AB - A method was developed for the determination of trace amounts of acrylamide (AA) in foods. The method includes the addition of 13C-labeled acrylamide-1-13C (AA-1 13C) as an internal standard, extraction with water, bromination, clean-up with a Florisil cartridge column, dehydrobromination and GC/MS analysis in the selected ion monitoring (SIM) mode. Bromination of AA to 2,3-dibromopropionamide (2,3 DBPA) was done using potassium bromide and potassium bromate under an acidic condition. 2,3-DBPA was converted to 2-bromopropenamide (2-BPA) by dehydrobromination with triethylamine before GC/MS analysis. The recoveries of AA from spiked potato chips, corn snack, pretzel and roasted tea were 97-105%, and their relative standard deviations were 0.8-3.9%. The detection limit of AA in foods was 9 ng/g. The method was applied to thirty-one foods purchased from retail markets. AA was found in potato chips at the level of 466-3,340 ng/g, and in other foods at the level of ND-520 ng/g. PMID- 12635342 TI - [Contents of plasticizers in cap-sealing for bottled food and their migration into food]. AB - Plasticizers in cap-sealing for bottled foods were analyzed. Twenty-three domestic samples and 80 imported samples, a total of 103 samples, were tested. Among them, 93 contained chloride, and 62 contained di(2-ethylhexyl) phthalate (DEHP), diisodecyl phthalate, O-acetyl tributyl citrate, diacetyllauroyl glycerol, di(2-ethylhexyl) adipate, diisononyl phthalate or dicyclohexyl phthalate. Twelve samples with DEHP-containing caps were further tested for plasticizer levels in the foods. Higher DEHP was detected in oily and fluid food specimens. However, the intake of plasticizers calculated based on usual food consumption did not exceed the tolerable daily intake level in any of the cases. A food sample containing the highest level of DEHP was stored under various conditions, and DEHP that migrated from the cap-sealing into the food was determined. Shaking the bottles increased migration of DEHP into foods. PMID- 12635343 TI - [Toxicity and tetramine contents of salivary glands from carnivorous gastropods]. AB - Salivary glands from 29 species of marine carnivorous gastropods in nine families were examined for lethal activity against mice and tetramine content. Mouse lethality was assayed by intravenous injection of buffer extracts into mice, and was detected in 14 species. Heat-stability tests confirmed that toxins in four species were thermolabile, while those in eight species were thermostable. Based on the tetramine contents determined by the colorimetric method using methanolic extracts, the thermostable toxins in seven species (Neptunea eulimatalamellosa, N. vinosa, N. arthritica, N. bulbacea, N. intersculpta f. pribiloffensis, N. intersculpta f. frater pilsbry and Hemifusus tuba) were considered to be tetramine contained at high levels (more than 900 micrograms/g salivary gland), but that in one species (Buccinum opisthoplectum) appeared to be a low-molecular weight compound differing from tetramine. It is interesting that one (Hemifusus tuba) of the seven species containing high amounts of tetramine belongs to the family Melongenidae, although the other six Neptunea species are members of the family Buccinidae, as expected from previous studies. PMID- 12635344 TI - Determination of seventeen pesticide residues in agricultural products by LC/MS. AB - Residues of 17 pesticides in agricultural products were determined by LC/MS with an atmospheric pressure chemical ionization (APCI) interface in both positive and negative ion modes. Pesticides were extracted with acetonitrile, and the extracts were cleaned-up with a primary and secondary amine (PSA) mini-column eluted with acetone-hexane (1:1). Rice, orange and potato were spiked with the 17 pesticides at 0.1 microgram/g and analyzed by the proposed method. The average recoveries of these pesticides usually ranged from 70 to 98% and the relative standard deviations were usually around 10%. These results suggested that LC/MS with APCI could be used to determine the residue levels of the 17 pesticides in these crops. PMID- 12635345 TI - [Protozoa and food hygiene]. PMID- 12635346 TI - [Acrylamide in foods]. PMID- 12635347 TI - [Hormonal interactions of parathormone and calcium metabolism]. AB - The possible hormonal interactions of parathormone and extracellular calcium level with other endocrine systems were studied. Primary hyperparathyroidism was used first as a clinical model, in which hypercalcemia and normocalcemia occurs before and after surgery, respectively. An increased activity of renin aldosterone system related to parathormone was found in hyperparathyroidism, and surgery resulted in a small decrease in blood pressure. This change was accompanied by a significant decrease in the activity of the renin-aldosterone system indicating the cessation of the secondary hyperaldosteronism. The role of a relative hyperinsulinism, occurring in hyperparathyroidism, in the pathogenesis of hypertension was not proved. The basal and stimulated secretion of thyreotrophin, the basal growth hormone level, and the stimulated prolactin secretion increased after surgery. Follicle stimulating hormone and luteinizing hormone secretions remained unchanged. The results suggest that extracellular calcium may reversibly modify the secretion of certain anterior pituitary hormones and their stimulus-induced responses. In the second disease, growth hormone deficiency syndrome, studied, long-term growth hormone replacement therapy results in significant but transient changes in bone metabolism: calcium , alkaline phosphatase-, and phosphate levels increase until 6 to 18 months as compared to the initial values; then these parameters decrease to the baseline level. Parathormone decreases until the first year then returns to the baseline level. Osteocalcin shows similar temporary changes. In spite of the above transient changes, osteodensity increases after 12 months of treatment, and further improvement can be seen after 18 and 24 months, i.e. GH treatment exerts a biphasic effect on bones; resorption increases first followed by an increase in formation. Based on the above results, it can be concluded that both parathormone and extracellular calcium are able to influence the secretion of certain hormones; and--as it is shown in growth hormone replacement therapy--other hormones may cause certain effect on them, too. The better understanding of these interactions may result in a better understanding of the pathomechanism of certain diseases and the improvement of their treatment. PMID- 12635348 TI - [Cut-off values for the diagnosis of obesity and hypertension based on epidemiologic measurements in Hungarian youth]. AB - INTRODUCTION: A representative nutritional hygienic survey was performed in a population of secondary school students (ageing from 15 to 18 years) between the years of 1997 and 2000. Herewith, the authors give an account on the anthropometric and blood pressure studies. METHODS: SAMPLING: the representative sampling sites were selected randomly. In the capital three-stage, in the counties four-stage stratified groups were assigned for the studies. The data for 3071 girls and 3274 boys were evaluated. Anthropometric parameters determined were as follows, body mass, body height, waist and hip circumference. The body mass index (BMI) and the waist/hip ratio was calculated. Blood pressure values were recorded using OMRON M4 automatic device. Repeated blood pressure measurements were performed twice at least > or = two weeks apart if the systolic blood pressure was 135 and/or the diastolic pressure was > or = 85 mmHg. Statistical analysis was performed using SPSS for Windows 9.0. RESULTS: Out of the anthropometric parameters the gender and age-specific percentile distribution is given in regard to body mass, body height, body mass index (BMI), waist circumference, and waist/hip ratio. There was a strong relationship between the waist circumference and BMI. Elevated blood pressure values were found at the first recording in 14.1% of boys and in 2.5% of girls. In students who presented three times, the incidence of high blood pressure was 7.5% in boys and 1.1% in girls. Only 1% of students was aware of his or her hypertension. CONCLUSIONS: The representative survey provides reference values concerning body mass, body height, BMI, waist circumference, waist/hip ratio and blood pressure values in the age group of 15 to 18 years. It also provides data for the prevalence of hypertension and gives cut off values for the diagnosis of leanness, overweight and obesity. PMID- 12635349 TI - [DNA chips for gene expression analysis and their application in diagnostics]. AB - Due to large genome projects more and more genetic information and more and more data about genetic background of various diseases can be obtained. By means of microarrays thousands to hundreds of thousands of molecular analysis can be performed at a time, rapidly, under identical conditions, hereby microarrays provide a solution to continuously increasing throughput demands from large genomic, pharmacogenetic and proteomic programmes. Microarrays offer unprecedented opportunities for qualitative and quantitative analysis of gene expression, understanding gene function and detection gene alterations, polimorphisms. Microarray expression profiles will define stages of different tumor types and new prognostic subgroups in cancer and other diseases. For individual patients, more precise diagnosis and risk assessment based on expression profiles are already achievable, leading to more accurate determination of prognosis and more individually tailored treatment. Microarrays will play an important role in diagnostics of infectious diseases and identification of novel diagnostic and therapeutic targets. Wide-spread application of microarrays in clinical practice is not typical today, but their production on an industrial scale and the increased information provided by microarrays should assure their entry into clinical practice as very powerful, sensitive, specific and rapid tools of diagnostics of various diseases. PMID- 12635352 TI - [ "Nosologic crisis" in the era of molecular medicine]. PMID- 12635350 TI - [The first clinical experiences in Hungary with a new effective antibiotic (linezolid) effective against Gram-positive infections]. AB - INTRODUCTION: The occurrence of gram-positive infections caused by multiresistant organisms has increased significantly in general, particularly among the surgical patients, and only a few effective antibiotics are available. A new and effective, synthetic antibiotic, against gram-positives is the oxazolidinon group, that electively inhibits bacterial protein synthesis in the early phase. Linezolid, the first member of this group to be used in clinical practice is the linezolid was studied. AIMS: The purpose of this study was to evaluate the effectiveness and safety of linezolid in the gram-positive infections of surgical patients. METHODS: A 3rd phase, clinical trial was conducted at the Semmelweis University Ist Surgical Department in the period of 1999-2001. Twenty-one patients with gram-positive infections were enrolled to this study. The mean of age was 57 years. Patients were selected for linezolid treatment in whom the conventional anti-gram-positive antibiotic therapy caused difficulties. RESULTS: Sixteen patients out of 21 recovered, one patient was cured clinically, but not microbiologically, and one case showed microbiological cure with clinical failure. In one case the methicillin resistant Staphylococcus aureus carriership was cured. Two cases had a fatal outcome. The causes of death were mediastinitis plus pneumonia in one case, and diffuse peritonitis with renal insufficiency in the other. Withdrawal from the study occurred in one case, due to drug intolerance. CONCLUSIONS: The linezolid administration proved to be safe and effective even in those cases, in which either hypacusis or decreased renal function persisted, or oral intake was advantagous. Contraindication of linezolid therapy did not occur. Few side effects were observed. If the infection was polymicrobial, the linezolid could be combined with other antibiotics. Further investigations are mandatory to evaluate the role of linezolid in the treatment of methicillin resistant Staphylococcus aureus carriership. PMID- 12635351 TI - [Excellent quality in health care]. PMID- 12635353 TI - [Zinc and nociception: a pharmacological approach]. AB - In the last decade, neurophysiological research regarding nociceptive mediation seems to suggest a possible new approach in pain management--zinc applicability. Immunohistochemical studies prove that zinc can be found in central nervous system, inside structures active in nociception. Zinc stabilizes primary afferent C fibers, nervous fibers involved in nociception. However, this phenomenon is probably only true for extreme zinc concentrations--excess or deficit. Electrophysiological studies indicate that zinc inhibits NMDA receptors, while potentiating the activity of non-NMDA receptors. All these are arguments supporting the existence of a role of endogenous zinc regarding pain transmission. PMID- 12635354 TI - [18F FDG PET-Applications in Oncology]. AB - In the first part our intention was, essentially, to present the particularities of glucose tumoral cells metabolism, PET components, the synthesis of 18F FDG and the detection of unknown cancers. This second part makes reference about mainly types of tumors who benefit by FDG-PET indications. Clinical PET has a rapid growth because of its use in cancer diagnosis and management. According with published studies all over the world, the sensibility and specificity of FDG-PET, noninvasive method, is higher than that of the conventional methods like CT, IRM, ultrasonography. PET is en excellent detection method of most of common cancer types and depends not on the histological neoplasm type; the more aggressive is the tumor, more it will uptake the radiotracer. The cost is significant, so the indications must be very precise: evaluating the malignity of solitary pulmonary nodules, evaluating the recurrences of melanoma, colon cancer diagnosis, differentiation between recurrent brain tumor and radiation injury, differential diagnosis of the benign lymph and malign lymph nodes, staging of Hodgkin's and non-Hodgkin's lymphoma, evaluation the response to therapy. Because the PET images are difficult to interpret, appears the necessity of correlation with anatomic images: this was the fusion images beginnings (the PET and CT images combination); now the physiologic information has precise anatomic localization. The growing of this method is very probably, both using 18F FDG -thanks to its highly favorable physical characteristics- and other new radiopharmaceuticals. The clinical cases that illustrate the applications are investigated at CERMEP, Lyon, France. PMID- 12635355 TI - [New inflammation modulator interleukins . Therapeutic implications]. AB - A lot of different leukocyte recruit soluble factors are implicated in both acute and chronic inflammatory processes with a higher expression of the cellular adhesion and chemoattraction molecules. Soluble factors implicated in these processes include: lipid inflammatory metabolites, arachydonic acid derivatives (prostaglandines, leucotrienes, lipoxines, PAF), the three soluble proteases waterfall events/substrates (coagulation, complement, kinines), nitric oxide, as well as cytokines implicated in acute or chronic inflammatory and cellular immune reactions. Recently, new inflammation modulator cytokines were described: IL-20, IL-21, IL-22 and IL-23. The aim of this work was to present some theoretical aspects regarding the biological effects of these new cytokines implicated in inflammation and the pharmacological influence of the processes mediated by them. Particularly IL-20 is implicated in psoriasis pathology. IL-21 and IL-15 are important in NK cells differentiation. IL-22 regulates the IL-4 production from Th2T cells. IL-23 stimulates IFN gamma production and proliferation in PHA blast T cells, as well as in CD45RO (memory) T cells. PMID- 12635356 TI - [Cardiovascular risk after renal transplantation(II): ischemic heart disease, hypercholesterolemia, hyperhomocysteinemia and smoking]. AB - Cardiovascular disease is the main cause of death in end-stage renal failure treated by hemodialysis or peritoneal dialysis. Though reduced in renal transplant recipients compared to the dialysis population, an excess cardiovascular mortality is still present after transplantation. The authors are reviewing the main data on mortality in the renal transplant population, focusing on major risk factors: ischaemic heart disease, hypercolesterolemia, smoking, hyperhomocysteinemia. The presence of these factors and the extent of cardiac and vascular abnormalities in the dialytic patient are closely related to outcomes in the post-transplant period. It is thus mandatory to approach and minimize all these in the dialytic and even predialytic period of chronic renal failure in order to reduce renal transplant mortality in patients with functioning grafts. Finally, an algorythm in managing cardiovascular disease pre- and post transplantation is proposed. PMID- 12635357 TI - [Prophylaxis of colorectal cancer with non steroidal anti-inflammatory drugs- a new reality?]. AB - The use COX-2 inhibitors induces regression of adenoma polyps and disrupts the sequence adenoma colorectal carcinoma. So that, this can be used in chemoprevention of colorectal cancer and also in cancer localised in other segments of the digestive tract. So far there is no agreement regarding the beginning of the treatment, the minimal efficient dose, the span of time required for chemoprevention. As the current studies in this field are quite encouraging we believe that in the next future COX-2 inhibitors could be used not only in chemoprevention but also in tumoral regression, and as a codrug in chemiotherapy of colorectal cancer. PMID- 12635358 TI - [Anatomical and surgical considerations concerning the segmentation of the pancreas]. AB - Pancreatic vascularization, especially the right segment (the head of the pancreas) has great surgical importance, due to its anatomical variety. From this practical point of view, establishing the type of the patient's pancreatic vascular pattern is highly recommended not just diagnosis purposes, but especially for its indispensable role in surgical decision. Selecting the appropriate technique, based on the imagistic evidences must consider the anatomical studies, which propose a systematic vision of the cephalic area vascularization, detecting two major types of arterial distribution. The first type is especially anastomotic; meanwhile the second one is terminal, these two varieties leading to a different surgical technique, best adapted for avoiding hemorrhages or ischemic necroses. Between the classic two segments of the pancreas (cephalic and caudal parts) there is a less vascularized area, which includes just an intersegmentary artery and the pancreatic duct, surgically sectioned during the separation operation of the two pancreatic segments. Vascular variety of the pancreas may be related to both embryological development of the pancreatic tissue, and that of the abdominal aortic branches. PMID- 12635359 TI - [ Etiopathogenesis of autoimmune thyroiditis]. AB - Human autoimmune thyroid diseases include both autoimmune hypothyroidism (thyroiditis) and autoimmune hyperthyroidism (Graves' disease). Either of the two feature profuse thyroidal infiltrates of CD4+ and CD8+ T-cells and a biased intrathyroidal T-cell-receptor repertoire. Despite strong epidemiologic evidence in favor of a genetic component in the etiology of autoimmune thyroiditis, few hereditary risk factors have been consistently identified, which include a well characterized series of HLA genes. These may account for the progression from a harmless autoimmune response characterized solely by production of autoantibodies to thyroglobulin to pathogenic autoimmunity where injury occurs to the thyroid cells. We hereafter summarize the role of inherited risk factors along with that of environmental risk factors. It was suggested that iodine increase the autoantigenic potency of thyroglobulin, a major pathogenic antigen in the induction of autoimmune thyroiditis. The clinical entities included together as autoimmune thyroiditis is shortly reviewed, along with the presentation of the common pathogenetic pathways. Unique features of each member of the group are further emphasized. PMID- 12635360 TI - [Phyllodes tumor. The experience of the 4th Obstetrics-Gynecology Department]. AB - The experience of the IVth Obstretrics Gynecology Department. Between the 1st of January 1994 and the 1st of November 2001 587 operations have been performed as treatment of breast tumors in the IVth Obstetrics and Gynecology Department. The phyllodes tumor has been diagnosed in 11 cases (1.8% of the benign breast masses). The genesis is unknown. Local trauma, pregnancy, lactation, high estrogenic levels have been suggested to have an influence upon the tumoral development. The most common aspect is a round, hard, unpainful breast mass. The diameter is variable (1 to 45 cm). The mammogram shows a well defined, high density, smooth contour opacity. A four degree classification has been described. Specific evolution features may involve local recurrences and metastasis. The surgical treatment is mandatory but radiotherapy, chemotherapy and endocrine treatment protocols have been brought up. PMID- 12635361 TI - [Hyperprolactinemia and pregnancy]. AB - Prolactin is a polipeptidic hormone secreted by the anterior pituitary gland, whose main role consists in the stimulation of lactation in the postpartum period. The increasing of prolactin secretion can be physiological (pregnancy and lactation) or pathological (hypothalamic and pituitary diseases, iatrogenic, etc.). The suppression of the hypothalamic-pituitary-gonadal axis and the resistivity of the ovary and gonadotropin action are induced in hyperprolactinemia, the amenorrhea and lack of ovulation being the result. Infertility associated with hyperprolactinemia is reversible with treatment, irrespectively to the type of treatment (radical or medical). Lowering of prolactin levels to normal or near normal is often necessary to permit ovulation. Dopamine-like drogs, as bromocriptine, appear to be safe for the developing fetus, at lest when the treatment is interrupted in the first trimester of pregnancy. PMID- 12635362 TI - [Renal cell carcinoma- diagnosis by computed tomography]. AB - In approximately 10% of cases, renal cell carcinoma (RCC) could present as a fluid- filled cystic mass. There are three mechanisms by which RCC may become cystic: extensive cystic necrosis, intrinsic cystic growth and origin from the epithelium lining a simple renal cyst. Simple renal cysts are very common. Uncommonly these cysts are complicated by hemorrhage, infection and possibly ischemia. The goal of the radiologist in evaluating these cystic lesions is to distinguish malignant neoplastic cystic masses from non-neoplastic complicated cysts so that appropriate management can be undertaken: RCC is best treated by surgical excision while non-neoplastic complicated cysts do not require surgery. The radiologic findings in these cystic masses which must be carefully evaluated include calcification, abnormal density, septations, nodularity, wall thickening and enhancement. PMID- 12635363 TI - [Staging and diagnosis in accidental hypothermia]. AB - Core body temperature below 35 degrees C is defining arbitrarily hypothermia. There is no worldwide consensus concerning the staging and resuscitation strategies in such a vital emergency, not even in rewarming strategy. Accidental hypothermia has its own "survival chain", modifying some steps or the timing in the common cardiopulmonary resuscitation protocol, according to some particularities of the metabolism in such an accident. Taking into account the two major events during hypothermic conditions (ventricular fibrillation and coma), we have proposed a better borderline between the three severity classes, based on clinical, paraclinical and prognostic arguments. The interest in this special environmental emergency situation is coming not only from its incidence, but especially from its particular long time period in which the resuscitation maneuvers could be effective, so that a literature review mixed with our practical observations may be of didactical and legal benefit also. PMID- 12635364 TI - [Establishing the ideal relationship between the dentist and the patient during therapeutical decisions]. AB - When it is regarded from the marketing and the management point of view, the relationship between the dentist and the patient can be theoretically superposed over the relationship between a professional and his client. The essence of this kind of relationship is to reveal to the client aspects that can not be achieved without the help of a professional. The accomplishment of these goals implies that both persons (the client and the professional) make decisions. The aim of this study is to observe and analyze the decisional capacity and how this can affect the relationship between the dentist and his patient. PMID- 12635365 TI - Hepatitis C virus serotypes in Romanian patients with chronic hepatitis and cirrhosis. AB - OBJECTIVES: To determine the prevalence of hepatitis C virus (HCV) serotypes in Romanian patients with chronic hepatitis infections and the association between them and disease's severity. PATIENTS AND METHODS: Anti-HCV antibodies were first investigated by an indirect ELISA (Murex IIIrd generation). From 55 patients positive for anti-HCV, 22 patients (11 female and 11 male) with chronic active hepatitis (CAH--6 cases), chronic persistent hepatitis (CPH--11 cases) and cirrhosis (C--5 cases) were tested for HCV serotypes using an indirect ELISA (Murex HCV Serotypes 1-6 Assay). The average age for the selected group was 47.2 years (range 27 divided by 68). RESULTS: In the first study from our region concerning this subject we have found the following serotypes distribution: serotype 1--68.18% (6/11--CPH; 4/6--CAH; 5/5--C); serotypes 2, 3, 4, and 6--4.54% for each of them (all were CPH cases); untypable--13.65% (2 with CAH and 1 CPH respectively). CONCLUSIONS: (1) The serotype 1 seems to be predominant in our region in patients with chronic HCV infections and it is associated with the most severe cases. (2) The diversity of serotypes is higher compared with previous study group results (e.g. in haemodialized patients the serotype 1 was determined in almost 100% cases--unpublished data). (3) Untypable strains (13.65%) require the sequence analysis for HCV genome characterization. PMID- 12635366 TI - [Effects of polyamine-loaded liposomes on liver mitochondrial permeability transition]. AB - The objective of the present study was represented by the effects of polyamineloaded liposomes on hepatic mitochondrial permeability transition (MPT). MPT was appreciated through the swelling of the isolated guinea-pig liver mithocondria induced by Ca2+, at 540 nm, using a diode-array 8452 UV-VIS spectrophotometer and a General Purpose software (Hewlett-Packard). Polyamines encapsulated in liposomes might inhibit the appearance of MPT induced by Ca2+, directly proportional to electrical charge: spermine > spermidine > putrescine > cadaverine. Thus, together with some previous data, it was observed that spermine may modulate hepatic MPT from the exterior, as well as from the interior of mitochondria. Moreover, spermidine and putrescine may also modulate MPT from the interior of mitochondria at low doses, and from the exterior only at high doses. The changes in mitochondrial membranes lipid composition (as done by control liposomes) are slightly influencing the MPT. The increase in membrane rigidity through the use of 40% cholesterol-enriched liposomes is drastically decreasing the appearance and development of MPT. PMID- 12635368 TI - [ANCA's- approach, advantages, limits]. AB - OBJECTIVES: The aim of this paper is to present the experience of the Laboratory of the Department of Histology in establishing the presence of ANCAs, in cases which cover an extremely large and diverse pathological area. The investigations were made following the request of some clinical medical services, ANCAs being a complementary exam, with an important role in the orientation and/or confirmation of the diagnosis. METHODS: There were investigated 77 cases with clinical diagnoses ranging in the connective tissue diseases field, mainly SLE and vasculitides, and in the renal diseases field, mainly glomerulonephritis. There was performed an indirect immunofluorescence technique with a personal method, involving the usage of blood smear instead of neutrophils concentrate, human anti IgG monospecific fluorescent conjugate and serum from the patients. RESULTS AND DISCUSSIONS: There was obtained a comprehensive survey of the presence of ANCAs in the study group. The interpretation of the results was completed within the general clinical background and in association with the physician. There is discussed the significance of the data in comparison with the literature. CONCLUSIONS: The ANCAs research may become a monitoring method of different cases, offering therapeutic and prognostic efficiency elements. PMID- 12635367 TI - [Phytoestrogens role in bone functional structure protection in the ovariectomized rat]. AB - Effects of soy protein diet on bone formation and density were evaluated in ovariectomized rats as a model for postmenopausal women. Twenty-seven 9-month-old rats were assigned to 3 treatment groups for the 9-week study: sham-surgery (Sh, n = 9); ovariectomy (Ovx, n = 9); ovariectomy + soy diet (OvxS, n = 9). Rats had free access to an AIN-93 M diet or AIN-93 M diet with 7% soy protein concentration and water. At sacrifice, rear legs were removed, and the right femur and tibia were cleaned manually. Serum alkaline phosphatase, a marker of bone formation, was measured colorimetrically. Bone density was measured using Archimedes' Principle. Alkaline phosphatase activity was greater in OvxS (114 +/- 19 U/L) and Ovx (128 +/- 26 U/L) compared to Sh (110 +/- 22 U/L). Femur bone density was greater for OvxS (1.520 +/- 0.02 g/cc) compared to Ovx (1.510 +/- 0.017 g/cc), but not to Sh (1532 +/- 0.025 g/cc). Tibia bone density was greater for OvxS (1.560 +/- 0.019 g/cc) compared to Ovx (1.553 +/- 0.015 g/cc), but not to Sh (1566 +/- 0.03 g/cc). In conclusion soy protein diet increased the rate of bone formation and bone density in some bones, suggesting that may help prevent bone loss in postmenopausal women. PMID- 12635369 TI - Histological aspects of the pulmonary territory as seen in an experimentally ovalbumin induced inflammation in guinea pigs. AB - The microscopic morphology of the respiratory territory was investigated on sections of pulmonary tissue and bronchioalveolar lavage liquid (BAL) that were stained with Giemsa, PAS and trichrome solutions. As a result of the induced pathological conditions, the following histological images were encountered: normal histological aspect of the bronchoalveolar territory was seen in the groups nebulized with 0.9% NaCl or sensitized after i.p. administration of ovalbumin (OA); macrophage cells influx in both tissue samples and BAL in animals nebulized with OA; after sensitization with OA followed by nebulization with OA, the same sequence of events as in atopical asthma was reproduced, including loss of epithelial structure and the appearance of mast cells and basophils in the alveolar territory. Hydrocortisone hemisuccinate, used to treat asthma attacks, causes a similar histological aspect as in the untreated group. Cells with intact basophilic granules were seen in the hypersensitized group under ketotiphen protection. PMID- 12635370 TI - [The implications of thromboembolism in chronic heart failure]. AB - The goal of investigation was to determine whether long-term anticoagulant therapy influences the mortality rate in CHF. The method consisted in the calculation of the annual death rate of the patients with CHF class III-IV NYHA: group A (controls)--who did not receive anticoagulant or antiplatelet therapy; group B--treated with Acenocumarol or Aspirin. The results show in group A, which included 150 patients, during the 5-year interval under study 30 deaths, representing an annual death rate of 4%. In group B, which included 325 patients of which 75 treated with Acenocumarol and 250 patients with aspirin, 20 deaths were recorded during the same 5-year interval, representing an annual death rate of 1.2%. Thus, the mortality risk proved to be 70% lower in group B than in the control group. It came out that the main mechanism of death in CHF is thrombembolism and in this circumstance anticoagulant or antiplatelet therapy would be essential. PMID- 12635371 TI - [Permanent cardiac pacing for chronic symptomatic atrioventricular block in uremic hemodialysed patients. A prospective study]. AB - Though sudden cardiac death accounts for as much as 15% of all cause mortality in uremia, reports concerning advanced A-V block, requiring permanent cardiac pacing in end-stage renal disease (ESRD) hemodialysed (HD) patients are very few. This is the first long term prospective study reporting on systematic permanent pacemaker implantation, in a cohort of ESRD patients from a single HD unit. Between 01/06/1997 and 30/12/2001, 396 pacemakers were inserted for advanced, symptomatic A-V block in our institution, including 5 in ESRD, HD patients (M/F- 4/1, age 47-73, M +/- SD--61 +/- 12 years) from a single dialysis center, treating 137 patients during the study period. Thus, the incidence and prevalence of A-V defects treated by permanent pacing in uremic patients was 0.81% and 3.65% respectively. Conversely, the incidence and prevalence of ESRD treated by hemodialysis, among patients with advanced A-V conduction disturbances, requiring permanent pacing were 0.28% and 1.26%. Mitral valve calcifications were present in all patients; 3 subjects also had extensive aortic valve calcifications. Left ventricular hypertrophy (echocardiographic Framingham criteria) was present in 4 patients, but the systolic function (ejection fraction and fractional shortening index) was normal in all cases, although a clinical picture of chronic heart failure was seen in 3 subjects preoperatively. A-V conduction defects were attributed to extensive metastatic calcifications, involving the cardiac squeleton, consecutive to severe hyperparathyroidism and inadvertent use of calcitriol and calcium carbonate as phosphate binders. No technical difficulties, short or long-term complications related to pacemaker implantation (4 VVI and 1 VVD devices) were encountered. Acute threshold and sensing values were similar with those of non-uremic patients. During follow-up, one patients died from a non cardiac death. If optimal hemodialysis is provided, benefits of permanent pacing are equal in uremic or non uremic patients and pacemaker implantation should be instituted as a prompt life-saving method in all dialysis patients with chronic symptomatic advanced A-V blocks. PMID- 12635372 TI - [Treatment with angiotensin conversion enzyme inhibitors in heart failure]. AB - The appearance of the Angiotensine Conversion Enzyme Inhibitors represents one of the most important events in cardio-vascular therapy, being much utilized in the last few years. The present study had as a main aim the evaluation of the benefit of the treatment with ACE Inhibitors in heart failure. Analyzing the results obtained, we can appreciate that ACE Inhibitors have a real benefit in patients with heart failure, improving the performance of physical effort and the life quality. PMID- 12635374 TI - [The segmentary colectomy or the left hemicolectomy for sigmoid cancer]. AB - The aim of this study was to show the benefits of segmentary colectomy (Raybard) for tumors located on sigmoid colon. A retrospective study was carried on 90 observations performed in the III-rd Surgery Clinic, Iasi between 1994 and 1998. Types of surgery performed were: left hemicolectomy (7 observation); segmentary colectomy (54 observation); left lower colectomy (17 observation); Hartmann operation (12 observation). CONCLUSIONS: This analysis showed the same survival for segmentary colectomy and left hemicolectomy. We consider segmentary colectomy the basic operation for this type of cancer. PMID- 12635373 TI - [Undesirable effects and interaction to angiotensin converting enzyme inhibitors therapy]. AB - By their intervention upon the mechanisms regulating the vascular tone, renal plasma flow and direct actions of chemical structures, angiotensin-converting enzyme (ACE) inhibitors may determine undesirable effects. These effects formed the object of a 5-year retrospective study (1995-1999) carried out at the IIIrd Medical Clinic of Iasi. During this interval ACE inhibitors were administrated to 2178 patients with hypertensive and coronary disorders or heart failure of various causes. Different generations of ACE inhibitors were used, but captopril, enalapril and lysinopril were the most commonly administered. Undesirable effects were recorded in 161 patients (7.3%). The following side-effects, single or associated, were recorded: 38 patients (23.6%) had increasing blood pressure proportional with ACEI dose, 80 patients (49.7%) had decreasing blood pressure at low doses ACEI, 23 patients (14.4%) had kidney failure, 2 patients (1.2%) had both increasing blood pressure and kidney failure, 3 patients (1.9%) had both decreasing blood pressure and kidney failure, 6 patients (3.8%) had dry cough, one patient (0.6%) had kidney failure with decrease blood pressure and allergic dermatitis, 4 patients (2.4%) had allergic dermatitis, and 4 patients (2.4%) had headache, vertigo, paresthesia. CONCLUSIONS: The treatment with ACE inhibitors has to be carefully initiated under strict clinical and biological monitoring, preferably in hospital setting. No drug associations that favor the undesirable effects of ACE inhibitors were reported. PMID- 12635375 TI - [Our experience in the surgical treatment of the arterial hypertension due to endocrine disturbances]. AB - This study is trying to expose our experience in the surgical treatment for adrenal tumors which produce arterial hypertension, experience gained over a period of 25 years. There were 93 adrenalcctomies carried out for these kinds of lesions, out of a total of 125 surgical procedures on the adrenal glands. Best results were obtained for phaeochromocytomas and also for tumors that produce primary hyperaldosteronism. Real advances have been done in the field of adrenal glands diseases by diagnosis and treatment but minimal invasive surgery is best used for all adrenal lesions but phaeochromocytomas in which this approach has to be very well judged. PMID- 12635376 TI - [Cajal interstitial cells identification]. AB - Cajal interstitial cells are cells that are found in the abdominal digestive system wall, between neurons and smooth muscular tissue. They are considered to be pacemakers for slow intestinal waves. The paper discuss about the electron microscopic identification of Cajal interstitial cells in the rat small intestine wall, cell morphology, placement of these cells in the muscular layer and the relation between Cajal interstitial cells and the components of the nervous plexus. Fragments of rat small intestine have been prepared for electron microscopy examination. Cajal interstitial cells have been found in different locations: in the circular muscular layer, around the nervous nodes and between the muscular layers (longitudinal and circular). The main morphologic characteristic of these cells is the aspect of cytoplasm, with numerous vacuoles and long extensions, some of them very thin, with a tendency to divide. Some Cajal interstitial cells form a network that surrounds the nervous nodes. Other form junctions with the muscular cells and with the interstitial neurons. PMID- 12635377 TI - [Quantitative histopathological criteria in predicting the giant bone cell tumors's grading]. AB - The giant bone cell tumour is a benign osteolitical tumour of spongious tissue. The evolving characters claimed the identification of the high-risk tumours for improving the prognostics. There were used samples of primitive tumours from 69 patients, paraffin included and coloured H&E and then examined with interactive digital video software. The quantitative standard measurements of giant cells (aria, perimeter and diameter), stereology (percentual volumes of cells, blood vessels and stroma) and proliferative activity assessment were made on the representative sections. The dimensions of the giant cells are higher in the Grade I and are lower in Grade II and III, with a dimensional variability. Bone giant cell tumours with mitotic rate less 1/mm2 were exclusively nonaggressive. Quantitative studies reveal the morphopathological changing specific for the grading and evolving forms of giant bone cell tumours. The stereology and mitotic activity index are essential predictive indicators and may be used in early detection of agressiveness and malignancy. PMID- 12635378 TI - [Evolution of superolateral surface of the cerebral hemisphere on 16-21 weeks fetus]. AB - Edification of neocortex is accompanied by the development and growth of the cerebral hemisphere, both processes being part of the more complex one, known under the name of telencephalization. The expression of this process is more acute on the superolateral surface of the cerebral hemisphere that expands laterally by growth of the frontal, temporal and parietal lobes. We describe the modifications of shape and deepness of the lateral cerebral fossa including the stages of its closure. We consider this event as the beginning of the appearance of gyri and sulci on the superolateral surface of the cerebral hemisphere. PMID- 12635379 TI - [Surgery experience regarding the pathology of the greater omentum]. AB - This study followed the evaluation of the surgical pathology of the greater omentum. From the casuistry of Urology Surgery Clinic in Hospital 2 Iasi, we have selected 1500 omentum pathologic specimens (inflammatory, traumatic, tumoral) that were investigated macroscopically and microscopically. The specimens were harvested during 1993-2000. Acute inflammatory involvement of the greater omentum recognizes a primitive etiology in 8-10% cases and secondary one in 90-92% cases; these lesions are associated to some inflammatory processes of peritoneal viscera by direct or indirect mechanisms. Non-tumoral pathology was signaled in 201 cases, malignant and benign tumors were found in 198 cases. Acute catarrhallis and phlegmonous omentitis were identified in: peritonitis by digestive perforation (240 cases), bladder perforation (12 cases), broken pyosalpinx (18 cases), acute calculosic colecystitis (260 cases), hernia (75 cases), strangled eventration (65 cases). The conclusion of our study is that there is a specific pathology for the omentum but also the latter is largely involved in the emergency and iterative abdominal surgical pathology. PMID- 12635380 TI - [Fungal infection in AIDS patients. Consideration for 10 cases]. AB - Clinic, etiologic and therapeutic study of fungic infection in AIDS patients. The retrospective clinical study of the 10 AIDS patients admitted between 01.01.1994 and 31.12.2000 in Infectious Diseases Clinical Hospital of Iasi. The majority of the cases (7) were registered in the last 3 years: 4 children and 6 adults. HIV infection was known only for 4 patients according to their history, and for the other 6 patients this diagnosis was made concomitantly of fungic infection diagnosis. The disease onset was insidious in 7 cases (2 children and 5 adults) being characterized by manifestations of nervous system involvement and consciousness disturbances (6 cases). The etiological diagnosis was established for alive patients by cerebrospinal fluid culture and hemocultures and Cryptococcus neoformans was isolated for 9 patients and Candida to another one. Fungic infection lead to meningeal injuries to 6 patients and the systemic one in for 4 patients. Although, under antifungical and antiretroviral therapy, the disease was lethal for 4 patients. The fungic infection recrudesces in AIDS patients, possessing systemic or/and meningeal manifestation, long evolution that can be lethal for a lot of them. PMID- 12635381 TI - [Possibilities of metallic-ceramic link optimization by electrodeposition]. AB - To reach a good link between the metallic framework and the ceramic element of a bridge represents one of the most important aim of the clinical and technical steps of the dental treatment for partial edentulous patients. The purpose of this study was represented by the analysis of the electrodeposition method in enhancing of the metalo-ceramic link. We used an electrogalvanic bath inside which we introduced a metallic framework which has been improved by electrodeposition of a thin layer of gold. The results showed a stronger and more stable link between the metallic framework and the ceramic element. PMID- 12635382 TI - [Dicephalic conjoined twins-a case report]. AB - Presentation of a case of dicephalic conjoined twins, rare malformation with an incidence of 1:50000. Before dissection, the specimen was investigated by CT, x ray and ultrasound. Dissection was performed under Zeiss surgical microscope and the images were taken with a Sony video line. Specimens for histologic examination were collected. The parents refused genetic examination. The complex study of the specimen allowed us to make the following diagnosis: dicephalus dibrahii dipus conjoined twins. In the past 20 years no such case was reported in Romania. An etiological diagnosis is impossible. Embryologically, we support the fusion theory (Winslow) promoted by O'Rahily (1998): gemellarity disturbance during the first week of pregnancy. PMID- 12635383 TI - [Method and optical device for exactly hinge axis localization tested on original phantom]. AB - The quickly and exactly method of location of the hinge-axis is based on the moire effect, produced between two superposed optical nets, one of it being moved. A face bow is fixed on the mandible to the occlusion rim. Two optical devices are mounted on the bottoms of the face bow. Two other optical nets are applied on the skin, corresponding to the TMJ. During the opening movements of the mandible when a pure rotation of the mandibular condiles take place on the TMJ, the optical nets are displaced and me exactly register the place corresponding to the rotation axe and we can note on the skin the hinge axis. The checkout was done using a phanthom achieved in our department, consisting of a skull made up of polyestheric resins and an elastic element made up of PVC. PMID- 12635384 TI - [Treatment of the displaced distal humeral fractures- the original AO procedure by means of two reconstructive plates]. AB - The aim of this paper is to assess the actual method of surgical treatment of the displaced fractures of the distal humerus, that is the AO technique, by means of 2 reconstructive plates. Until recently, we were very keen on using the Lecestre type plate placed on the lateral pillar of the distal humerus. Since September 2001 we have managed to treat these types of fractures in the manner they are treated all over the world. Thus, the arch of motion of the elbow is improved both in terms of flexion and extension. Moreover, the 2 reconstructive plates can be properly molded on the surface of the bone entailing a firmly stabilized distal humerus. PMID- 12635385 TI - [ First studies of etnoiatry in Moldavia]. PMID- 12635386 TI - [An anatomy lesson in Romanian sculpture at the end of XIX th century ]. PMID- 12635387 TI - Local anaesthesia--before cocaine. PMID- 12635388 TI - The efficacy and cost-effectiveness of prophylactic 5-hydroxytryptamine3 receptor antagonists: tropisetron, ondansetron and dolasetron. AB - There are currently three 5-hydroxytryptamine3 (5-HT3) receptor antagonists available in Australia. In this randomized, double-blind, parallel group study the prophylactic antiemetic effect of a single dose of tropisetron 2 mg, ondansetron 4 mg or dolasetron 12.5 mg was compared after major gynaecological surgery. One hundred and eighteen patients (group T n = 42; group O n = 36; group D n = 40) were evaluated for nausea, vomiting, recovery characteristics and satisfaction for 24 hours postoperatively. A cost-effectiveness analysis was performed. Rescue antiemetic, prochlorperazine 12.5 mg i.m., was given if vomiting occurred more than 10 minutes after arrival in the recovery room. If prochlorperazine was ineffective one hour after administration, droperidol 1 mg i.v. was given. There were no significant differences between groups for the incidence of vomiting during consecutive epochs until 24 hours postoperatively or overall (57%, 75% and 72.5% for groups T, O and D respectively, P = 0.18). The incidence and number of rescue antiementic treatments for nausea or vomiting were similar. The incidence of nausea and the overall and interval nausea scores were similar except for lower "worst nausea" score in group T between 12 and 18 hours (P = 0.02). Recovery times, satisfaction and cost per patient did not differ between groups. We conclude that the risk of postoperative nausea and vomiting remained high in this setting despite 5-HT3 receptor antagonist prophylaxis and that the choice between these agents should be based on the lowest available acquisition cost. PMID- 12635389 TI - Midazolam pretreatment reduces etomidate-induced myoclonic movements. AB - During induction of anaesthesia with etomidate, myoclonic muscle movements are frequent. In this study, pretreatment with a small dosage of etomidate or midazolam was compared with placebo for the prevention of myoclonic muscle movements. Sixty patients, premedicated with oral midazolam, were pretreated in a randomized double-blinded fashion with etomidate 0.05 mg/kg i.v., midazolam 0.015 mg/kg i.v. or normal saline i.v. (placebo) in three groups of 20 patients each. The pretreatment was followed after 90 seconds by etomidate 0.3 mg/kg i.v. One minute after onset of hypnosis, induction of anaesthesia was completed with sufentanil and rocuronium. From the time of pretreatment to completion of anaesthesia, patients were observed for myoclonic muscle movements by a single physician, blinded to group allocation. Myoclonic movements were graded on a scale of 0 to 3. The incidence of myoclonic movements was significantly lower in patients pretreated with midazolam (4 of 20) compared with placebo (18/20) (P < 0.01). Midazolam 0.015 mg/kg i.v., administered 90 seconds before induction of anaesthesia with etomidate, is effective in reducing etomidate-induced myoclonic muscle movements. PMID- 12635390 TI - Increased B-type natriuretic peptide (BNP) level is a strong predictor for cardiac dysfunction in intensive care unit patients. AB - Patients admitted to an Intensive Care Unit (ICU) frequently have underlying cardiac dysfunction. Early interventions are sometimes difficult to initiate because of diagnostic uncertainty as to whether cardiac failure is present As B type natriuretic peptide (BNP) has been shown to be increased in cardiac dysfunction, we sought to demonstrate whether BNP can be used as a screening tool for cardiac dysfunction in patients admitted to ICU. All patients admitted to a combined medical and surgical ICU over a four-week period were included in the study. BNP was measured on the point of admission using a hand-held meter. Clinicians were blinded from the measurement when diagnoses were made as to whether or not the patients had clinically significant cardiac dysfunction. Patients with cardiac dysfunction had a significantly higher level of BNP when compared to the non-cardiac dysfunction group: 516 +/- 385 pg/ml (n = 26) v 67 +/ 89 pg/ml (n = 58) (P < 0.0001) A BNP cut-off value at 144 pg/ml exhibited a 92% sensitivity, 86% specificity and 96% negative predictive value. The sensitivity improved to 96% when the analysis was confined to patients > or = 55 years. At this cut-off value, BNP is a strong predictor of cardiac dysfunction. BNP measurement offers a rapid and affordable way to screen for cardiac dysfunction in patients admitted to ICU. An increased BNP level warrants further cardiac investigations so as to implement early interventions for cardiac decompensation in ICU patients. PMID- 12635391 TI - Intermittent enteral nutrition--a comparative study examining the effect on gastric pH and microbial colonization rates. AB - In a prospective randomized study we aimed to examine the effect on gastric pH and microbial colonization of enteral nutrition (EN) delivered both by intermittent and continual infusion. Forty-three mechanically ventilated patients were randomized to receive EN by one of three methods, intermittent or continual gastric or continual jejunal. We sampled gastric aspirate for pH and culture in all patients at 0600 hours and pH in gastric intermittent feeders at 2200 hours daily. Patients were studied for 12 days or until extubated. Data was obtained on 41 patients, of whom 73% had a diagnosis of trauma. Median APACHE II score was 17 and ventilation time 11 days. Twelve patients received gastric continuous, 15 gastric intermittent and 14 jejunal nutrition. No significant difference was observed between the three groups with regard to median 0600 pH (P = 0.16). This was lowest in the jejunal group (3.2) followed by the gastric intermittent group (4.0) and then gastric continuous group (5.0). With overnight cessation of EN in the gastric intermittent group, the median pH fell from 5.2 at 2200 to 4.0 at 0600 (P = 0.01) with no effect on gastric colonization. The probability of gastric colonization over time however was significantly lower in the jejunal group compared with the two gastric groups (Log rank test, P = 0.02). These results demonstrate that in a patient population consisting predominantly of trauma, overnight cessation of EN made no overall difference to 0600 gastric pH or colonization rates. The preservation of pH and lowered colonization in those given jejunal feeding may have implications in the pathogenesis of ventilator associated infection and may warrant further study using larger patient numbers. PMID- 12635392 TI - Comparison of peripheral and central venous pressures in critically Ill patients. AB - We conducted a prospective study to determine the relationship between central (CVP) and peripheral (PVP) venous pressures in critically ill patients. CVP and PVP were measured on five different occasions in 20 critically ill patients in the intensive care unit. Results showed that the mean difference between PVP and CVP was 4.4 mmHg (95% CI = 3.7 to 5.0). However, PVP might be 1.9 mmHg below (95% CI = 0.7 to 3.1) or 10.6 mmHg above (95% CI = 9.4 to 11.8) the CVP. The mean difference between changes in PVP and corresponding changes in CVP was 0.3 mmHg (95% CI = -0.1 to 0.7). The actual change in PVP could be 3.0 mmHg below (95% CI = 2.3 to 3.7) or 3.6 mmHg above (95% CI = 2.9 to 4.3) the change in CVP. Overall, the direction of change in PVP (rise or drop) predicted a same direction of change in CVP with an accuracy of 78%. Changes in PVP > or = 2 mmHg predicted a change in same direction of CVP with an accuracy of 90%. The direction of changes in CVP > or = 2 mmHg were predicted by the direction of change in PVP with an accuracy of 91%. We conclude that PVP measurement does not give an accurate estimate of the absolute value of CVP in individual patients. However, as changes in PVP parallel, in direction, changes in CVP, serial measurements of PVP may have a value in determining volume status and guiding fluid therapy in critically ill patients. PMID- 12635393 TI - The effects of a one unit blood donation on auto-haemodilution and coagulation. AB - The effect of haemodilution on coagulation has been extensively investigated. We investigated auto-haemodilution following a 10% blood loss (480 ml) and its effect on coagulation. Ten healthy, unstarved volunteers were enrolled. One unit of blood was taken from each volunteer. Concurrently blood was taken from the opposite arm prior to and immediately after the blood donation, and at 1, 2, 4 and 6 hours. It was tested for thrombelastography, haematocrit and endorphins. There was a significant decrease in r-time from the control sample to the sample taken immediately post blood donation. This value returned to baseline at 1 hour post donation and did not change again. There were no other significant changes in thromboelastographic parameters. Fractional plasma noradrenaline changes were significantly raised at 1 hour post donation (P = 0.048), returning to baseline by 2 hours post donation. The haematocrit showed a rapid (approximately 4%) fall during donation followed by a slow, but progressive decrease over six hours, falling by a mean of 8.3% from pre-donation values. A state of relative hypercoagulability is found immediately after a rapid 10% loss in circulating blood volume. This may be related to the rapid immediate haemodilution. It is unlikely that the sympathetic response to blood loss plays a role. However, after the initial drop, slow restoration of circulating blood volume by autodilution takes six to eight hours, and is not associated with enhanced coagulation. Of interest is that a 10% blood loss in a healthy person does not require volume replacement. PMID- 12635395 TI - Dimensions of double-lumen tracheobronchial tubes. AB - The diameter of the left main bronchus is the determining dimension when selecting the size of a left tracheobronchial (double-lumen) tube for lung separation. However, this information is not given by any manufacturer, either on the tube or in the package insert. This paper describes the lengths and diameters of the deflated bronchial cuff segment of left tracheobronchial tubes in common use. One hundred and seventy-one left tracheobronchial tubes ranging in size from 28 to 41 nominal French gauge from four manufacturers were measured. There was wide variation between tubes of the same nominal size from the same manufacturer. For tubes of the same size from the same manufacturer, the diameter of the segment with the deflated bronchial cuff varied by more than 1 mm in diameter in some instances. The diameter of the bronchial cuff segment did not consistently decrease as the nominal size decreased even for the same manufacturer. There was major overlap in diameters of the bronchial segments between Fr 41, Fr 39, and Fr 37 tubes from most manufacturers, so that some of the Fr 39 tubes have a bronchial cuff segment diameter as much as 0.5 mm larger than the Fr 41 tube. It is concluded that the current French gauge markings on left tracheobronchial tubes are of very limited value in determining the appropriate size to be selected for a patient. More accurate and consistent dimensions of tracheobronchial tubes are required to improve clinical selection. PMID- 12635394 TI - Avoiding inadvertent epidural injection of drugs intended for non-epidural use. AB - Inadvertent administration of non-epidural medications into the epidural space has the potential for serious morbidity and mortality. The aim of this study was to collate reported incidents of this type, describe the potential mechanisms of occurrence and identify possible solutions. We searched medical databases and reviewed reference lists of papers retrieved, covering a period of 35 years, regarding this type of medication incident. The 31 reports of 37 cases found is likely to represent a gross underestimation of the actual number of incidents that occur. "Syringe swap", "ampoule error", and epidural/intravenous line confusion were the main sources of error in 36/37 cases (97%). Given that no effective treatment for such errors has been identified, prevention should be the main defence strategy. Despite all the precautions that are currently undertaken, accidents will inevitably occur. We have identified areas for systemwide change that may prevent these types of incidents from occurring in future. PMID- 12635396 TI - Compensating for the effect of inlet gas temperature on heated humidifier performance. AB - The humidity output of heated humidifiers may be compromised by inlet gas temperatures exceeding approximately 26 degrees C, with humidity dropping below the recommended levels for intubated patients. A new version of the Fisher & Paykel MR850 humidifier claims to deal with this problem by offering a humidity compensation option. The present study tested this feature by measuring humidity output using the gravimetric method and a hygrometer at different inlet gas temperatures (16.6 degrees C to 40.0 degrees C) with compensation on and off. It was found that the compensation is effective in maintaining humidity levels despite high inlet gas temperatures. PMID- 12635397 TI - Preparing a new generation anaesthetic machine for patients susceptible to malignant hyperthermia. AB - Anaesthetic machines are prepared for use with patients who are susceptible to malignant hyperpyrexia (MH) by flushing with oxygen at 10 l/min for ten minutes to reduce the anaesthetic concentration to 1 part per million (ppm) or less. Anaesthetic workstations are now often used in place of traditional machines. Workstations have greater internal complexity, and it is not known if they can be made safe for susceptible patients by flushing with oxygen. We used a high sensitivity infrared gas analyser to measure the washout of isoflurane from five Datex-Ohmeda workstations. Measurements were then repeated with a patient breathing circuit. Isoflurane washout occurred in an exponential manner. The time to reach a concentration of 1 ppm at the fresh gas outlet was 17 +/- 7 minutes, and all machines had reached less than 2 ppm by ten minutes. The washout of isoflurane from the machine and patient breathing circuit was much slower than from the machine alone, with a concentration less than 2 ppm reached only after 30 minutes. We conclude that the Datex-Ohmeda workstation can be prepared for use in MH susceptible patients by flushing with oxygen at 10 l/min for ten minutes. Flushing of the patient breathing system is not straightforward, and we recommend using a clean T-piece circuit. If the circle system and ventilator are required for anaesthesia, we suggest using new breathing hoses, rebreathing bag and soda lime cartridge, and ventilating an artificial lung for 30 minutes with a fresh gas flow rate of 10 l/min and tidal volume of 1 litre. PMID- 12635398 TI - Patients' knowledge of and attitudes towards awareness and depth of anaesthesia monitoring. AB - Awareness during anaesthesia is uncommon (approximately 0.1%), but causes significant anxiety, dissatisfaction and morbidity for patients. Several electroencephalographic monitors hold promise as monitors for awareness. We therefore conducted a survey to evaluate patients' knowledge of and attitudes towards awareness and monitors of anaesthetic depth. Two hundred consenting, preoperative patients completed a seven-item questionnaire. The median number of previous operations was 2 (inter-quartile range, 1-5). Thirteen patients reported an experience which they thought might be awareness (2% of operations performed on the cohort). Only 56% of patients had heard about awareness before and many (35%) of these had heard about it in the media. Many (35%) were uncertain about what might cause awareness. Many (42.5%) were anxious about awareness: female sex and not having heard about awareness before were significant predictors of anxiety. Nevertheless only 34% were willing to pay for a proven awareness monitor if they were at low risk and only 50% if they were at high risk. Perceived risk and a previous awareness experience were significant predictors of willingness to pay for awareness monitoring. PMID- 12635399 TI - Predictors of preoperative anxiety in children. AB - This study aimed to identify factors contributing to anxiety at induction of anaesthesia in children. One hundred and twenty children aged five to twelve years and scheduled for surgery requiring general anaesthesia were included. Children were interviewed and assessed prior to surgery. Parents completed anxiety measures prior to surgery and were interviewed after the induction of anaesthesia. The level of children's anxiety was determined at the time of induction of anaesthesia by the modified Yale Preoperative Anxiety Scale. Factors associated with increased levels of anxiety in the children included increased number of people in the room at induction of anaesthesia; longer waiting time between admission at the hospital and induction of anaesthesia; negative memories of previous hospital experiences; and having a mother who does not practise a religion. Suggestions for implementation of the findings and for future research are provided. PMID- 12635400 TI - Consequences of syringe size sensor malfunction in a modern infusion pump. AB - Prompted by an actual case of potentially life-threatening infusion pump malfunction, we investigated the effects of wire breakage(s) within the syringe size sensor circuit in a Graseby 3400 infusion pump. The circuit wires within the sensor were systematically broken. The syringe sizes recognised by the sabotaged circuit and the actual sizes of syringes inserted into the pump were compared. Thirty-eight per cent of the possible wire breakages resulted in a smaller syringe size being recognized, causing the infusion rate to be too fast, and 38% of the possiblewire breakage resulted in a larger syringe size being recognized, causing the infusion rate to be too slow. The volume delivered for each different size of Terumo syringe as a function of distance travelled by the plunger was measured. The errors ranged from 0.4 to 2.6 times that of the expected rate. Only 1.3% of the possible wire breakage(s) were recognised as errors by the pump. The infusion rates were not affected in 22.5% of the cases. Wire breakage within the syringe size sensor in infusion pumps is yet another potential source of infusion error, with important safety implications. PMID- 12635401 TI - A prospective clinical audit of central venous catheter use and complications in 1000 consecutive patients. AB - We conducted a prospective audit of central venous catheter (CVC) use in 1000 consecutive patients to better define the rates of postoperative complications (particularly vascular perforation) and the pattern of CVC and pulmonary artery catheter (PAC) usage (particularly the number of lumens inserted and utilized). Details of CVCs, complications, and the number of lumens in place and used, were recorded daily until all CVCs were removed. A total of 1546 CVCs and 223 PACs were placed in study patients. Two non-fatal perforations occurred: a perforated right atrium in a patient who received an Arrow triple-lumen CVC (previously reported), and a perforated pulmonary artery in a patient upon withdrawal of a Baxter PAC. The risk per patient of any CVC-related perforation was 0.2% (95% confidence interval (CI): 0.02% to 0.7%). The rates of CVC-related sepsis and local infection were 3% (95% CI: 2% to 4%) and 2% (95% CI: 1% to 3%) respectively. At the peak of CVC use (day 1 in the ICU) the overall number of lumens placed was significantly correlated with lumens used (r = 0.53), endorsing clinical judgement in the anticipation of the needs of the patient. The modal number of lumen uses in adults and children was four. However, in children, fewer catheters were inserted per patient than in adults (1.28 vs 1.63, P = 0.01), and placed lumens were used more intensively (P < 0.001). Data appear to justify the routine selection of a triple-lumen CVC in adult patients, but not of a quad lumen CVC. PMID- 12635403 TI - Anaesthesia for caesarean section in a patient with spinal muscular atrophy. AB - This report describes a 33-year-old primigravid woman with spinal muscular atrophy Type III (Kugelberg-Welander syndrome). Elective caesarean section was performed at 38 weeks gestation under spinal anaesthesia. The implications of spinal muscular atrophy for anaesthesia for caesarean section are described. PMID- 12635402 TI - Papaverine angioplasty to treat cerebral vasospasm following traumatic subarachnoid haemorrhage. AB - The management of vasospasm associated with traumatic subarachnoid haemorrhage presents many challenges. We present a 20-year-old male admitted after sustaining a closed head injury complicated by a Fisher grade III traumatic subarachnoid haemorrhage. Despite treatment with intravenous nimodipine he developed a delayed ischaemic neurological deficit due to cerebral arterial vasospasm. The vasospasm was successfully managed with serial papaverine angioplasty. PMID- 12635404 TI - Anti-factor Xa monitoring of anticoagulation during cardiopulmonary bypass in a patient with antiphospholipid syndrome. AB - The activated clotting time (ACT) may be an unreliable monitor of coagulation for patients with the antiphospholipid syndrome. We describe a patient with antiphospholipid syndrome in whom adequate anticoagulation during cardiopulmonary bypass was confirmed by monitoring both the ACT and anti-factor Xa levels. The cardiopulmonary bypass was uneventful, and there were no thrombotic or bleeding complications. The use of anti-factor Xa levels provided confirmation of adequate anticoagulation (and reversal of anticoagulation) that was not possible using the ACT alone. PMID- 12635405 TI - Angioedema of the tongue due to acquired C1 esterase inhibitor deficiency. AB - We describe the management of an 83-year-old woman who presented with upper airway obstruction due to angioedema of the tongue. Following definitive airway management, investigation showed a diagnosis of acquired C1 esterase inhibitor deficiency (acquired angioedema) that was considered to be subsequent to haematological malignancy. Resolution of the macroglossia followed treatment with C1 esterase inhibitor concentrate, but the patient failed to wean from ventilatory support and died in the Intensive Care Unit. This case report highlights the potential for acquired angioedema to cause upper airway obstruction. The various treatment modalities for acquired C1 esterase inhibitor deficiency are summarized. PMID- 12635406 TI - Cantrell's pentalogy. PMID- 12635407 TI - Epistaxis and nasal tubes. PMID- 12635408 TI - Cross-reactivity between gelofusin and haemaccel. PMID- 12635409 TI - Dosage of dantrolene--"empiric" please! PMID- 12635410 TI - Epistaxis and nasal tubes. PMID- 12635411 TI - A case of aspiration using the proseal LMA. PMID- 12635412 TI - Gastrointestinal disorders during pregnancy. Preface. PMID- 12635413 TI - Abdominal pain during pregnancy. AB - Numerous medical, surgical, psychiatric, gynecologic, and obstetric disorders can cause abdominal pain during pregnancy. The patient history, physical examination, laboratory data, and radiologic findings usually provide the diagnosis. The pregnant woman has physiologic alterations that affect the clinical presentation, including atypical normative laboratory values. Abdominal ultrasound is generally the recommended radiologic imaging modality; roentgenograms are generally contraindicated during pregnancy because of radiation teratogenicity. Concerns about the fetus limit the pharmacotherapy. Maternal and fetal survival have recently increased in many life-threatening conditions, such as ectopic pregnancy, appendicitis, and eclampsia, because of improved diagnostic technology, better maternal and fetal monitoring, improved laparoscopic technology, and earlier therapy. PMID- 12635414 TI - Nutritional assessment and support during pregnancy. AB - This article thoroughly updates the authors' previous review of nutritional assessment and support during pregnancy. After briefly reviewing nutrient metabolism and requirements, the authors discuss the nutritional assessment of the pregnant woman and review the nutritional support principles in hyperemesis gravidarum and other conditions that can compromise the nutritional health of mother or fetus. PMID- 12635415 TI - The fetal safety and clinical efficacy of gastrointestinal endoscopy during pregnancy. AB - More than 12,000 pregnant patients in the United States per annum have conditions that are normally evaluated by EGD. More than 6000 pregnant patients in the United States per annum have conditions that are normally evaluated by sigmoidoscopy or colonoscopy. About one thousand more have symptomatic choledocholithiasis during pregnancy, which is a strong indication for endoscopic sphincterotomy in nonpregnant patients. Endoscopy during pregnancy raises the unique issue of fetal safety. Endoscopic medications comprise a significant component of fetal endoscopic risks. Safety of EGD during pregnancy has been examined in a case-controlled study of 83 patients, a mailed survey of 73 patients, and 28 case reports. Safety of sigmoidoscopy during pregnancy has been examined in a case-controlled study of 46 patients, a mailed survey of 13 patients, and 10 case reports. Safety of therapeutic ERCP during pregnancy has been analyzed in studies of 23, 10, 6, and 5 patients, and in 32 case reports. These studies suggested that EGD, sigmoidoscopy, and ERCP should be performed when strongly indicated: EGD for significant upper gastrointestinal bleeding, sigmoidoscopy for nonhemorrhoidal rectal bleeding, and ERCP for symptomatic choledocholithiasis when sphincterotomy is contemplated. PEG and colonoscopy are currently considered experimental during pregnancy because of insufficient data on fetal safety. Several cases of PEG and colonoscopy were successfully performed during pregnancy. Performance of endoscopy during pregnancy should increase with further technical refinements, and greater awareness of procedure safety. PMID- 12635416 TI - Gastrointestinal surgery and pregnancy. AB - Gastrointestinal disorders during pregnancy that require surgery often mimic the symptoms and signs of conditions that do not require surgery. Anatomic and physiologic changes of pregnancy can alter the usual clinical presentation of gastrointestinal disorders that require surgery. These alterations can be a challenge to diagnosis. Prompt treatment is critical to successful management. Most elective and urgent operations can be performed during pregnancy with minimal maternal and fetal risk. The condition of the mother should always take priority because proper treatment of the mother usually benefits the fetus as well. PMID- 12635417 TI - Nausea and vomiting during pregnancy. AB - The differential diagnosis of nausea and vomiting is extensive and the underlying cause can be difficult to diagnose. Treatment of nausea and vomiting also can be unsatisfactory because the available pharmacotherapy does not correct the fundamental underlying pathophysiologic abnormalities. In contrast, the diagnosis of nausea and vomiting of pregnancy generally is straightforward. Almost 80% of women have some degree of nausea in the first trimester of pregnancy, and the diagnosis of pregnancy is established easily by the patient or physician. The spectrum varies from mild nausea to hyperemesis gravidarum. Various treatment approaches are addressed. PMID- 12635418 TI - Gastroesophageal reflux disease during pregnancy. AB - Approximately two thirds of pregnant patients develop heartburn. The origin is multifactorial, but the predominant factor is a decrease in LES pressure caused by female sex hormones, especially progesterone. Mechanical factors play a small role. Serious reflux complications during pregnancy are rare; therefore EGD and other diagnostic tests are infrequently needed. Symptomatic GERD during pregnancy should be managed with a step-up algorithm beginning with lifestyle modifications and dietary changes. Antacids or sucralfate are considered the first-line medical therapy. If symptoms persist, H2RAs should be used. Ranitidine is probably preferred because of its documented efficacy and safety profile in pregnancy, even in the first trimester. Proton-pump inhibitors are reserved for the woman with intractable symptoms or complicated reflux disease. Lansoprazole may be the preferred PPI because of its safety profile in animals and case reports of safety in human pregnancies. PMID- 12635419 TI - Gastric and duodenal ulcers during pregnancy. AB - The frequency, symptoms, and complication rate of PUD seem to decrease during pregnancy. Yet clinicians often have to treat dyspepsia or pyrosis of undetermined origin during pregnancy because the frequency of pyrosis significantly increases during pregnancy, and clinicians reluctantly perform EGD during pregnancy for pyrosis to differentiate reliably between GERD and PUD. Dyspepsia or pyrosis during pregnancy is initially treated with dietary and lifestyle modifications. If the symptoms do not remit with these modifications, sucralfate or antacids, preferably magnesium-containing or aluminum-containing antacids, should be administered. Histamine2 receptor antagonists are recommended when symptoms are refractory to antacid or sucralfate therapy. Ranitidine seems to be a relatively safe H2 receptor antagonist. If symptoms continue despite H2 receptor antagonist therapy, the patient should be evaluated for possible EGD or PPI therapy. Pregnant women with hemodynamically significant upper gastrointestinal bleeding or other worrisome clinical findings should undergo EGD. Indications for surgery include ulcer perforation, ongoing active bleeding from an ulcer requiring transfusion of six or more units of packed erythrocytes, gastric outlet obstruction refractory to intense medical therapy, and a malignant gastric ulcer without evident metastases. PMID- 12635420 TI - Constipation, diarrhea, and symptomatic hemorrhoids during pregnancy. AB - Constipation, diarrhea, and symptomatic hemorrhoids are disorders common in the general population, particularly in women. These conditions, if mild, often are self-treated with various home remedies or nonprescription preparations. Few of these patients, moreover, are referred to gastroenterologists, as primary care providers generally are confident managing these conditions, unless they are severe, refractory to conventional management, or require additional diagnostic studies. PMID- 12635421 TI - Inflammatory bowel disease in pregnancy. AB - Fertility is affected in ulcerative colitis after surgery and in active Crohn's disease. Adverse fetal outcomes are not increased when IBD is quiescent. Active disease at conception increases the risk of adverse fetal outcomes. Most medications for IBD are safe during pregnancy and breastfeeding, with notable exceptions. Active disease is usually more deleterious than maintaining medical therapy. PMID- 12635422 TI - Colon cancer during pregnancy. AB - The medical community should become mobilized to diagnose colon cancer earlier in pregnancy to improve prognosis. The primary care physician or obstetrician should refer the pregnant patient with significant gastrointestinal complaints to the gastroenterologist for evaluation. Likewise, the gastroenterologist should be prepared to perform sigmoidoscopy, preferably without endoscopic medications, for significant lower gastrointestinal symptoms such as persistent rectal bleeding. Sigmoidoscopy is particularly sensitive in identifying colon cancer in pregnant patients because their cancers are usually distal and within reach of the sigmoidoscope. PMID- 12635423 TI - The irritable bowel syndrome during pregnancy. AB - The irritable bowel syndrome (IBS) is characterized by altered bowel habits and abdominal discomfort in the absence of organic disease. No markers exist for IBS, and the definition of IBS is based on the presence of specific symptoms. The Rome II criteria for defining IBS include abdominal pain or discomfort for 12 weeks or longer, which need not be continuous, over the past 12 months plus two of the following: (1) relief of discomfort with defecation; (2) association of discomfort with altered stool frequency; and (3) association of discomfort with altered stool form. Nine percent to 22% of the population report symptoms consistent with IBS. IBS is the most prevalent digestive disease, representing 12% of visits to primary care physicians and 28% of referrals to gastroenterologists. PMID- 12635424 TI - Pregnancy and liver disease. AB - Liver disease has an impact on women's health during pregnancy because of the complex interactions between the physiologic changes induced by pregnancy and the pathophysiologic changes of liver disease. In particular, liver diseases that predominantly afflict females, such as primary biliary cirrhosis and autoimmune hepatitis, pose a special problem for conception and management of pregnancy. Pregnancy, moreover, specifically is associated with several potentially life threatening liver diseases. This article reviews comprehensively the impact of liver diseases on pregnancy and of pregnancy on liver function and liver disease. PMID- 12635425 TI - Human immunodeficiency virus and pregnancy. AB - The recent spread of HIV infection into the heterosexual population in the United States, Europe, and Australia, as well as its earlier heterosexual presence in the developing world, has led to increased scientific and clinical attention to the role of HIV infection in pregnancy. In managing a pregnant HIV-positive woman, it is most important to treat the patient as someone who is HIV-positive rather than someone who is pregnant. Withholding antiviral or prophylactic therapies from the mother for fear of harming the child is not justified, because failure to treat the mother increases the fetal risk. The most important parameter to follow is the maternal plasma HIV-RNA level, and the most important treatment issue is to reduce this level because it is directly related to the risk of vertical transmission. PMID- 12635426 TI - Soliciting patient complaints to improve performance. AB - BACKGROUND: A study was conducted in 2000 to describe service quality problems in a large tertiary care teaching hospital and evaluate the effect of a pre discharge program for active complaint surveillance and resolution on patient satisfaction. METHODS: The pre-post intervention study with temporal controls was conducted at a tertiary care teaching hospital in St Louis. Eighty-four percent (1,023 of 1,218) of patients admitted to a general medical unit between October 2, 2000, and December 22, 2000, were interviewed by a patient advocate to identify and address patient complaints about service quality. Patient satisfaction was measured, using a validated instrument administered by telephone interview 7-10 days after discharge. RESULTS: The advocate completed 1,233 patient interviews and received 695 complaints about service quality. Half of the complaints concerned local unit care, most frequently delays in response to patient requests. Patients also complained about food, delays in admission and discharge, and inadequate communication about procedures. Concurrently, the hospital's formal reporting system received 12 complaints. Patients satisfaction scores were unchanged during the intervention. DISCUSSION: Active surveillance using predischarge patient interviews by a patient advocate identified many local and systemwide service quality problems in a large tertiary care teaching hospital that needed to be addressed to improve the quality of patient care. However, patient satisfaction scores were unchanged. PMID- 12635427 TI - How health care organizations can establish and conduct a program for a latex safe environment. AB - BACKGROUND: The advent of universal precautions brought about a dramatic increase in the use of natural rubber latex gloves, and reports of rubber latex allergies began appearing in the literature. In 1997 the Johns Hopkins Hospital created the interdisciplinary Latex Task Force to address the issue of creating, implementing, and evaluating a latex-safe environment. CONVERSION TO NONLATEX PRODUCTS AND EXAMINATION GLOVES: When suitable alternatives were available, all medical products that contained latex were to be removed from the hospital and nonlatex alternatives substituted. Latex medical gloves, especially powdered latex examination gloves, which were used in all patient care areas, were replaced by vinyl gloves. Yet because of the ongoing concern about strike-through and the minimal level of acceptance of fit with the vinyl gloves, the search for alternatives to the vinyl gloves continued. The task force recommended switching to nitrile examination gloves throughout the hospital. To facilitate the transition to another examination glove, new educational pamphlets about the nitrile gloves were developed. POSTSCRIPT: The switch to nitrile examination gloves was successfully completed, but conversion to nonlatex surgical gloves was less successful, with costs being the overwhelming impediment. Monitoring of latex-containing products and ongoing evaluations of alternatives are crucial in ensuring patient and health care worker safety. PMID- 12635428 TI - A multihospital safety improvement effort and the dissemination of new knowledge. AB - BACKGROUND: Research on the transfer of medical technology and guidelines suggests that this transfer is driven more by interpersonal relationships than by new research or available information and that it is inconsistent, largely unsuccessful, and strongly influenced by local factors. Yet studies of collaborative, multiple-hospital improvement efforts have shown these transfers to be effective for the specific microsystems participating in the project. The diffusion of medical innovations beyond the participating teams was studied during a 2000-2001 national collaborative safety improvement effort. METHODS: Twenty-two teams from Department of Veterans Affairs (VA) hospitals participated in a 9-month quality improvement project designed to improve safety in high hazard areas. Participating hospitals and other regional hospitals were contacted to determine the level of dissemination of information generated during and after the project. RESULTS: While the participating hospitals benefited from the quality improvement effort, changes were implemented only 9% of the time on other units within the hospitals and only 2% of the time in other regional hospitals. After 12 months, there was no implementation within participating hospitals, and other regional hospitals were implementing changes 10% of the time. DISCUSSION: Personal commitment from senior leadership, dissemination strategies that push information to clinicians, and monitoring of progress at the regional level are all needed for dissemination of complex medical information to occur. PMID- 12635430 TI - Using a market model to track advances in patient safety. AB - The author proposes a four-stage model that may help hospitals and other health care providers recognize and anticipate market drivers of patient safety. PMID- 12635429 TI - Do health care ratings differ by race or ethnicity? AB - BACKGROUND: There is growing evidence that Asians and Pacific Islanders perceive their health care more negatively than whites and other racial and ethnic subgroups. This study of differences in health care experiences by race and ethnicity was the first to use nationally representative data. METHODS: Data from the 1998 National Research Corporation Healthcare Market Guide survey were analyzed. A total of 120,855 respondents were included in the study. Four global satisfaction ratings (overall health plan satisfaction, medical care satisfaction, recommend plan to others, and intent to switch plans) and four composite measures (access to care, providers' delivery of care, customer service, and cost/benefits of care) were examined. RESULTS: Nonwhite survey respondents--particularly those in the other/multiracial and Asian/Pacific Islander groups--rated their health plan coverage and medical care lower than whites. DISCUSSION: The results of this study are consistent with those of other recent (and comparable) studies in which these racial and ethnic groups are represented. Most strikingly consistent, however, are the lower ratings of Asians/Pacific Islanders and the comparable (and higher) ratings (compared to whites) of African Americans. Why Asians/Pacific Islanders are considerably less satisfied with their medical care than all other racial and ethnic groups in the United States needs to be explored. Access to care and quality of care improvement efforts should be directed at all ethnic minority groups, particularly for limited English-speaking, other/multiracial, Hispanic, and Asian/Pacific Islander subgroups. PMID- 12635431 TI - Video-storytelling: a step-by-step guide. PMID- 12635432 TI - Why PAs should care about genomic medicine. PMID- 12635433 TI - What PAs need to know about smallpox. PMID- 12635434 TI - Should we measure CRP levels to assess cardiovascular risk? PMID- 12635435 TI - Current concepts in managing heart failure. Based on recent studies of beta blocker therapy, treatment guidelines have been revised for patients who have mild, moderate, or severe HF. PMID- 12635436 TI - Measuring compassion in physician assistants. AB - While compassion is generally agreed to be one of the professional qualities required of physician assistants (PAs), the concept has been remarkably difficult to define and measure. The authors describe a series of studies that generated preliminary information about the compassionate qualities of PAs. In an initial survey, PAs rated their own compassionate qualities, after which their patients assessed the same qualities in them. The patients validated three elements of compassion having to do with communication between PA and patient but failed to validate seven other items. Subsequent research dealt with a halo effect related to the wording of the questions and a confounding effect associated with the intimacy of the medical setting. PMID- 12635437 TI - How to incorporate genetic thinking into your practice. PMID- 12635438 TI - Advances in treating the infected total hip replacement. PMID- 12635439 TI - House calls in America. PMID- 12635440 TI - Future of Catholic health care in the new millennium. PMID- 12635441 TI - The end of Catholic health care? Or a new beginning? PMID- 12635442 TI - Migraine therapy selection not an isolated decision. PMID- 12635443 TI - Pharmaceutical case management helps Iowa Medicaid patients. PMID- 12635445 TI - Military reserve pharmacists provide vital services. PMID- 12635446 TI - Bedside calculation of stroke risk in patients with atrial fibrillation. PMID- 12635447 TI - Pharmacists and HIPAA. PMID- 12635448 TI - Compounding our problems--again. PMID- 12635449 TI - HIPAA and its impact on pharmacy practice. PMID- 12635450 TI - Galantamine hydrobromide: an agent for Alzheimer's disease. AB - The pharmacology, dosage, adverse effects, efficacy, and economics of galantamine hydrobromide are discussed. Galantamine hydrobromide is a tertiary alkaloid that has been extracted from plant sources and is now synthesized for use in the treatment of mild to moderate Alzheimer's disease (AD). Galantamine acts both as a reversible competitive inhibitor of acetylcholinesterase (AChE) and as an allosteric modulator of nicotinic acetylcholine receptors. The recommended starting dosage is 4 mg (as the hydrobromide) twice daily. The dosage should be increased in increments of 8 mg/day in two divided doses after four weeks at a given dosage until a maintenance dosage of 16-24 mg/day in two divided doses is reached. Adverse effects are primarily mild and cholinergic and include nausea, vomiting, diarrhea, and dizziness. Five large clinical trials demonstrated that galantamine is more effective than placebo in controlling the symptoms of mild to moderate AD. Optimal therapy appears to require early initiation of the drug and a dosage-adjustment period of eight weeks. In one study, galantamine delayed full time care by 10% and reduced the overall cost of care by $528. Because galantamine has not yet been compared directly with other AChE inhibitors, cost should be the principal factor weighed during formulary evaluations. Galantamine provides the clinician with another choice of an AChE inhibitor for use in treating AD. PMID- 12635451 TI - Use of antiresorptive drugs among older women: a case study in Wisconsin. AB - The factors contributing to the use of antiresorptive therapies (ARTs) and differentiating the ARTs used were examined. A list of names of community dwelling persons 65 years or older living in Wisconsin was obtained, and 2100 individuals were selected (1050 men and 1050 women) to receive the 10-page survey instrument. The survey requested information on health status, prescription drug use, preferences for using prescription drugs versus self-treatment strategies, demographic characteristics, and status and type of prescription drug and medical insurance. Only the responses from female respondents were evaluated. Descriptive statistics were calculated for each variable. The simultaneous relationships among all variables were modeled sequentially using a binary logistic regression model and a multinomial logistic regression model. Usable responses were received from 629 women, yielding an adjusted response rate of 59.9%; 169 (26.9%) reported current use of at least one ART, 110 (17.5%) reported using hormone replacement therapy (HRT), and 59 (9.4%) were using newer ARTs. The typical respondent did not use any prescription medicines for the treatment or prevention of osteoporosis, was married, and did not have a four-year college degree. Married, younger women with high medical care preferences scores were more likely to use antiresorptive agents. Married women were more likely to use HRT versus no treatment. A higher number of family practice physicians per 100,000 population reduced the likelihood of the use of any ART. Lower age was also related to the use of ARTs. Marital status, age, medical care preferences, prescription drug insurance, a regular source of care, and the diagnosis of osteoporosis were all significantly associated with the use of any antiresorptive agent. PMID- 12635452 TI - Stability of multidose, preserved formulation epoetin alfa in syringes for three and six weeks. AB - The integrity and biological activity of multidose, preserved formulation epoetin alfa stored in syringes at 2-8 degrees C were studied. Three independent 1.0-mL hubless syringes of epoetin alfa 20,000 units/mL were aseptically prepared and refrigerated for three and six weeks (a total of six syringes). Protein integrity was assayed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting, and glycoprotein detection. Biological activity was determined through a cell based proliferation assay. The presence or absence of microbial contamination was observed after a one-week culture. A multidose, preserved formulation of epoetin alfa that was opened only at the time of assay served as the reference standard. SDS-PAGE silver-stained gels and immunoblots demonstrated no evidence of erythropoietin degradation after three and six weeks of storage when compared with the reference standard. In addition, SDS-PAGE, immunoblotting, and direct glycoprotein detection found that protein glycosylation was unaffected by the storage. Student's t test detected no significant difference between stored samples and the reference standard in biological activity (p > 0.05). A culture of epoetin alfa in bacterial and eukaryotic cell growth media showed no evidence of contamination. The results suggest that epoetin alfa can be dispensed to patients in prefilled syringes every four to six weeks to coincide with their peritoneal dialysis schedule. The integrity and biological activity of 20,000 units/mL epoetin alfa in prefilled syringes remain intact after three and six weeks when stored at 2-8 degrees C. PMID- 12635454 TI - Effect of pharmacist counseling in patients hospitalized with acute exacerbation of asthma. PMID- 12635453 TI - Effects of pioglitazone on diabetes-related outcomes in Hispanic patients. PMID- 12635455 TI - Developing a pharmacy work force for the future. PMID- 12635456 TI - Precautions in use and handling of travoprost. PMID- 12635457 TI - Survey of clinical pharmacy services in a family medicine clinic. PMID- 12635458 TI - An adverse-drug-reaction-reporting system at a teaching hospital in India. PMID- 12635459 TI - Olanzapine and risperidone. PMID- 12635460 TI - [Advances in therapy of renal anemia: darbepoetin alfa]. PMID- 12635461 TI - [Alternative and natural science therapy forms: incompatible contrast]. AB - In spite of the accomplishments of science-oriented medicine, we are still confronted with a multitude of "alternative" or "complementary" therapies which claim to heal "holistically" without adverse effects. Common to alternative treatment methods, which are several or many centuries old, is the notion that a special force ("vis vitalis", "entelechy", "spiritual bioforce", etc.) is responsible for life that, ultimately, cannot be investigated. According to this perception, which is termed "vitalism", diseases are a result of changes in the immaterial force of life. Therefore, treatments have to be directed at this central regulator ("regulatory therapy"). The medicine that is based on the natural sciences, in contrast, presumes that all expressions of life including diseases are amenable to critical and rational analysis. According to this school of thought, a causal therapy can be derived only from a detailed knowledge of the various body functions down to the molecular level and the effects of drugs on these functions. Corresponding therapeutic theories have to be verified or falsified experimentally, excluding bias as much as possible. Using such "objective" methods, a statistical assessment of the beneficial and adverse effects of a treatment may be possible ("evidence-based medicine"). In the final analysis, an incompatibility of scientific concepts is at the heart of the controversy between alternative treatment methods and the medicine based on natural sciences. PMID- 12635462 TI - "Flower remedies": a systematic review of the clinical evidence. AB - BACKGROUND: Flower remedies (also called "Bach" flower remedies) are used by an increasing number of individuals, and many health claims have been made for them. No systematic review of flower remedies has so far been published. AIM: The aim of this systematic review is to summarise and critically analyse the data from all available controlled clinical trials of flower remedies. METHODS: Six databases were searched to identify all controlled clinical trials of flower remedies in humans for any medical condition. No language restrictions were applied. Key data were validated and extracted into table format according to pre defined criteria. Statistical pooling was not possible, and results were evaluated in narrative form. RESULTS: Four studies met the inclusion/exclusion criteria. Two trials suggested a positive outcome. Those studies that controlled for placebo-effects and minimised selection bias through randomisation failed to demonstrate effects beyond a placebo response. CONCLUSION: The hypothesis that flower remedies are associated with effects beyond a placebo response is not supported by data from rigorous clinical trials. PMID- 12635463 TI - [Treatment of renal anemia with darbepoetin alfa: results of an Austrian multicenter study]. AB - Darbepoetin alfa is a unique erythropoetic protein whose half-life is 3 times longer than that of recombinant human erythropoetin (rHuEPO). It corrects and maintains haemoglobin (Hb) concentrations at increased dosing intervals as compared to rHuEPO. The aim of this study was to evaluate the efficacy and safety of darbepoetin alfa administered as fixed unit doses. Haemodialysis patients (n = 250) maintained on stable rHuEPO treatment 2-3 times weekly (n = 200) were switched to darbepoetin alfa once weekly (QW). Treatment for patients on rHuEPO QW (n = 50) was changed to darbepoetin alfa every other week (Q2W). The route of administration (i.v. or s.c.) was kept unchanged. The dose of darbepoetin alfa was titrated to maintain Hb levels at 10-13 g/dL between baseline and the evaluation period (weeks 21-24; primary endpoint). There was no clinically relevant change in mean Hb levels between baseline (11.69 g/dL) and evaluation ( 0.28 g/dL (95% CI: -0.43; -0.13)). Mean weekly dose requirements of darbepoetin alfa decreased by 13.3% from 36.7 micrograms (95% CI: 33.9; 39.7) to 31.8 micrograms (95% CI: 28.7; 35.2). This decrease was more pronounced in patients receiving darbepoetin alfa i.v. (-18.4%) as compared to those receiving it s.c. ( 6.4%). Darbepoetin alfa was well tolerated. Overall safety data were consistent with those observed in other studies. These data confirm that unit dosing with darbepoetin alfa at increased dosing intervals and reduced dose effectively and safely maintains Hb levels in haemodialysis patients. PMID- 12635464 TI - Anti chlamydia antibodies in patients with thoracic and abdominal aortic aneurysms. AB - BACKGROUND: Chlamydia species are suspected of being involved in the pathogenesis and progression of aortic aneurysms. We investigated serum levels of Chlamydia antibodies in patients with thoracic aortic aneurysms (TAA) and abdominal aortic aneurysms (AAA) compared to levels in healthy individuals. METHODS: We included 35 consecutive patients with TAA, 42 patients with AAA and 42 age- and sex matched healthy controls in a case control study. Serum antibodies (IgM and IgG) against Chlamydia lipopolysaccharide (LPS), Chlamydia pneumoniae and Chlamydia trachomatis were measured by recombinant ELISA and quantified by measurement of optical density. RESULTS: Patients with TAA exhibited median immunoglobulin levels against Chlamydia LPS (IgM 0.090, IgG 0.266), C. pneumoniae (IgM 0.023, IgG 0.264) and C. trachomatis (IgG 0.247) comparable to those of healthy subjects [Chlamydia LPS IgM 0.209 (p = 0.1), IgG 0.301 (p = 0.2); C. pneumoniae IgM 0.051 (p = 0.07), IgG 0.516 (p = 0.1); C. trachomatis IgG 0.153 (p = 0.2)]. Patients with AAA had higher serum levels of IgG against Chlamydia LPS (0.560) compared to healthy individuals [0.301 (p = 0.04)], but no significant elevation of antibodies against C. pneumoniae [IgM 0.029 (p = 0.1), IgG 0.545 (p = 0.9)] and C. trachomatis [IgG 0.219 (p = 0.3)]. CONCLUSION: Thoracic aortic aneurysms were not associated with signs of Chlamydia infection or immunopathogenicity. In contrast, patients with abdominal aortic aneurysms exhibited elevated levels of immunoglobulin against Chlamydia LPS, reflecting an unspecific Chlamydia immunopathogenicity. However, elevated levels of antibodies against distinct Chlamydia species were also not found in AAA patients. PMID- 12635465 TI - Malignant non-Hodgkin's lymphoma of childhood and adolescence in Austria--therapy results between 1986 and 2000. AB - Between 1986 and 2000 183 Austrian children and adolescents with non-Hodgkin's lymphoma (NHL) and mature B-cell acute leukemia (B-ALL) were enrolled in 3 consecutive studies of the Berlin-Frankfurt-Munster (BFM) Group. In trial NHL-BFM 86, patients were stratified according to the histologic subtype and clinical stage. In the succeeding studies NHL-BFM 90 and 95, treatment stratification was additionally based on the speed of tumor response to therapy and for children with B-cell NHL/B-ALL also on the pre-therapeutic serum lactic dehydrogenase level. Event-free survival rates were 84% +/- 6% in trial NHL-BFM 86 (n = 39) and 86% +/- 4% in both trials NHL-BFM 90 (n = 67) and NHL-BFM 95 (n = 77). Patients with lymphoblastic lymphoma (mainly with T-cell phenotypes) had an excellent prognosis with an ALL-type chemotherapy regimen (n = 49; relapse, n = 1), whereas an intensive, short-pulse therapy delivered within a 2- to 4-month period was found to be highly efficacious in children with B-cell NHL/B-ALL (n = 114; relapse, n = 6; progression, n = 5). Patients with anaplastic large cell lymphoma (ALCL) who were treated with similar alternating short courses of multi-agent chemotherapy had a less good outcome (n = 20; relapse, n = 6, progression, n = 3). Children with B-cell NHL and B-ALL who failed initial therapy also had a dismal prognosis (10/11 patients died). Local radiotherapy as a part of lymphoma therapy was completely abandoned in study NHL-BFM 90 and surgical interventions were confined to specific situations such as complete resection in localized B cell NHL and ALCL, diagnostic biopsy and second-look operation. In conclusion, our results showed that the BFM treatment strategy for lymphoblastic lymphoma and B-cell NHL/B-ALL was highly successful in the majority of patients; however, optimal treatment for children with ALCL has not yet been defined. As a consequence, larger trials at an international level are necessary to find new prognostic markers that might define more precisely those patients who need further intensification of first-line treatment or novel therapy. PMID- 12635466 TI - Correlation of lipoprotein(a) levels and myocardial stress thallium scintigraphy pattern in different entities of CHD severity. AB - BACKGROUND: Higher levels of lipoprotein(a) confer an increased risk for coronary heart disease (CHD). Apo-E genotype (APO-E) also plays a role, the APO-E epsilon 4 allele being associated with CHD. Furthermore, higher Lp(a) concentrations are correlated with APO-E epsilon 4 allele presence. The study was performed to investigate the relationship of Lp(a) and APO-E with the functional status of coronary arteries as evaluated by myocardial scintigraphy. PATIENTS AND METHODS: We studied 70 patients (27 F and 43 M; mean age: 55 +/- 6 yrs.) consecutively referred for CHD, and 50 normal sex and age-matched controls. Total cholesterol, LDL-cholesterol, HDL-cholesterol, triglycerides, apo-AI, apo-B, Lp(a) levels (ELISA), Lp(a) isoforms (Immuno Blotting) and APO-E (PCR) were measured in all subjects. Only CHD patients underwent a myocardial tomographic stress thallium scintigraphy (201Tl-SPECT); the SPECT pattern was classified as follows: no perfusion defects (unstable angina group = 1), reversible defects at stress images (ischemia group = 2), fixed defects (infarction group = 3). RESULTS: Lp(a) medians were significantly higher than controls in group 1 (p < 0.05), 2 (p < 0.001) and 3 (p = 0.00). Low molecular weight isoforms (B, S1, S2) were significantly more frequent in all CHD-patients vs. controls (p < 0.05), whereas APO-E genotypes did not differ among controls and patients. Multiple regression analysis showed family history (p < 0.001) to be the only independent predictive variable of CHD severity correlated to the scintigraphic pattern. CONCLUSION: Among the considered biological parameters in our patients only Lp(a) plasma levels are related to the entity of ischemic cardiac wall damage as evaluated by 201Tl-SPECT. PMID- 12635467 TI - Step aerobic vs. cycle ergometer training: effects on aerobic capacity, coordinative tasks, and pleasure in untrained adults--a randomized controlled trial. AB - The purpose of the study was to evaluate the impact of step aerobic (StA) and cycle ergometer training (CET) on physical performance, coordination, and pleasure, during workout. Forty untrained persons (40-70 years) were randomly assigned to either of the two regimens. Prior to and after three months of training, we investigated the participants' physical performance with a cycle ergometer test and by testing coordinative tasks (upper extremities: tapping test; lower extremities: one-leg stance). After the training period, visual analog scales were used to evaluate personal assessment (pleasure, wellbeing, team spirit, interest in prolongation of training). StA increased the relative oxygen uptake at the anaerobic threshold (RVO2AT) while CET increased the relative maximal oxygen uptake (RVO2max) to a statistically significant extent. However, intergroup comparison failed to show group-specific differences. Concerning coordinative tasks, the members of the StA group achieved a significant time reduction for both hands' tapping test. However, only the improvement in left-handed tapping was significantly higher than that achieved by members of the CET group. Despite the absence of group-specific differences, CET members showed a statistically significant change when comparing the duration of pre- with post-training time for one-leg stance under proprioceptive conditions. Team spirit was significantly higher in the StA group than in the CET group. Except for the time reduction in left-handed tapping, the present study found no group-specific differences in physical performance and coordination. Participating in a StA class has a more cohesive effect on the individual members than attending a CET group. PMID- 12635468 TI - Simvastatin and pravastatin equally improve flow-mediated dilation in males with hypercholesterolemia. AB - BACKGROUND: Both simvastatin and pravastatin have been shown to improve endothelial function in patients with hypercholesterolemia. To our knowledge there has been no comparative study of these two HMG-CoA reductase inhibitors on endothelial dysfunction measured by flow-mediated dilation of the brachial artery in patients with hypercholesterolemia. METHODS: Fourteen middle-aged males with hypercholesterolemia (means +/- SD: total cholesterol 7.03 +/- 0.88 mmol/l, LDL cholesterol 5.02 +/- 0.63 mmol/l, HDL cholesterol 1.3 +/- 0.38 mmol/l and triglycerides 1.47 +/- 0.26 mmol/l) were randomised, after a 6 weeks' run-in phase with AHA step I diet treatment, to 12 weeks' treatment either with simvastatin or pravastatin. Both statins were given in a daily dose of 10 mg for 6 weeks, which was increased to 20 mg daily in patients who did not achieve an LDL-cholesterol goal of < 3.4 mmol/l. Endothelial dysfunction was measured as flow-mediated brachial artery dilation (FMD) using high resolution ultrasound. RESULTS: There were no significant differences between the drugs in reduction of total cholesterol, LDL cholesterol and triglycerides, or elevation of HDL cholesterol. FMD increased in the simvastatin group from 6.8 +/- 3.2 to 12.3 +/- 2.9% (p < 0.03) and in the pravastatin group from 6.3 +/- 4.8 to 13.3 +/- 4.7% (p = 0.001). The improvement in FMD was the same in both groups (p = 0.64) and did not correlate with changes of the lipid parameters measured. CONCLUSIONS: Both simvastatin and pravastatin reduce endothelial dysfunction to the same degree in patients with hypercholesterolemia, independently of changes in lipid parameters. PMID- 12635469 TI - Intravenous anti TNF-alpha antibody therapy leads to elevated triglyceride and reduced HDL-cholesterol levels in patients with rheumatoid and psoriatic arthritis. AB - BACKGROUND & AIMS: We investigated the effect of Infliximab, an anti TNF-alpha antibody, on plasma lipids and lipoproteins in patients with rheumatoid arthritis and psoriatic arthritis. METHODS: Five male and 10 female patients with a mean age of 56.7 years were included in this study. Seven of the patients were diagnosed with rheumatoid arthritis and 8 patients with psoriatic arthritis. All patients received infusions of 3 mg/kg Infliximab (at week 0, 2 and 6). Lipids, lipoproteins and standard clinical parameters were assessed at baseline (0 week), after 2 weeks, and in 4 patients after 6 weeks. RESULTS: There was a significant increase in triglyceride levels during treatment with Infliximab (112 +/- 48 versus 133 +/- 53 mg/dl, p < 0.01). In contrast, HDL-cholesterol levels were significantly lowered (56 +/- 12 versus 50 +/- 13 mg/dl, p < 0.006) by the treatment. There was no significant difference in total cholesterol (209 +/- 25 versus 205 +/- 36 mg/dl) or in LDL-cholesterol (131 +/- 24 versus 118 +/- 43 mg/dl) before and after treatment. Similarly, lipoprotein(a) levels did not alter during treatment (median: 1.1 versus 1.4 mg/dl). CONCLUSION: This study shows that intravenous Inflixmab therapy leads to changes in plasma lipid and lipoprotein levels in patients with rheumatoid and psoriatic arthritis and may result in a more atherogenic lipid and lipoprotein profile. Although larger patient numbers need to be studied to confirm our findings, these results suggest that lipid levels should be checked and monitored in patients receiving infliximab therapy, particularly in patients with vascular disease. PMID- 12635470 TI - Redistribution of corticotropin-releasing hormone receptor R2 using fluorescence immunohistochemistry in fetal membranes of women delivering preterm or at term. AB - OBJECTIVE: The aim of our study was to determine whether a difference exists in expression of corticotropin-releasing hormone receptor-R1 (CRH-R1) and corticotropin-releasing hormone receptor-R2 (CRH-R2) in fetal membranes of preterm and term women with or without labor. MATERIAL AND METHODS: Small pleces of fetal membranes were obtained from the placenta of each of forty patients undergoing cesarean section. Ten samples each were taken from preterm and term patients, with and without labor. Antibodies against CRH-R1/2 and CRH-R2 were used for localization by conventional fluorescence immunohistochemistry. The evaluation of staining was based on examination of the entire histologic section by three independent observers. RESULTS: In women at term without labor, CRH-R2 receptor was predominantly expressed in the amniotic epithelium and the amniotic mesenchyme. In laboring women at term, the expression of CRH-R2 receptor was shown in the chorionic mesenchyme and the cytotrophoblast cells, but no specific staining could be detected in the amniotic membranes. Changes in CRH-R2 receptor expression could not be demonstrated during preterm labor of early pregnancies. In preterm women, the antibody against CRH-R1/2 receptor detected additional signals in the amniotic mesenchyme and epithelium, suggesting expression of CRH R1 in these tissues. In women at term, the overlapping pattern of CRH-R1/2 was recognized in both the chorionic and amniotic mesenchyme, in contrast to the specific CRH-R2 staining, suggesting expression of CRH-R1 in the mesodermal cell compartments. CONCLUSION: At term, changes in CRH-R2 expression are directly related to the progression of normal labor; such changes were not observed during preterm labor of early pregnancies. The increased CRH-R2 expression in the chorionic mesenchyme may possibly provoke rupture of the membranes or at least play a role in some key regulatory events in the initiation of normal labor. The fact that this mechanism does not occur in preterm labor strengthens the hypothesis that onset of labor could be controlled by distinct mechanisms in preterm and term pregnancies. PMID- 12635471 TI - Cystic dystrophy of the duodenal wall in a heterotopic pancreas. AB - Cystic dystrophy of the duodenal wall in heterotopic pancreas is a rare disease affecting younger men in particular. On account of its difficult diagnosis and nonspecific signs, it was first described in 1970. A 51-year-old male patient was admitted for clarification of several months of severe pain in the upper abdomen accompanied by weight-loss, and due to ultrasonically established suspicion of carcinoma in the head of the pancreas. Clinical examination and laboratory tests were nonsignificant. Barium meal and esophagogastroduodenoscopy revealed a severe deformation of the duodenum. Biopsy of the duodenal mucosa detected moderate inflammatory changes. Ultrasound examination showed cystic changes in the duodenal wall and in the vicinity of the head of the pancreas. Only with the aid of endoscopic ultrasound, could bizarre cystic changes in the submucosa of the duodenal wall be detected, which was also confirmed by computer tomography. The signs of disease are nonspecific. Duodenal biopsy also does not reveal its cause since the typical lesions lie deeper in the submucosa or muscularis propria. Cystic changes with fibrosis, pancreatic ducts and lobi without anatomic or vascular connection to the pancreas are found. The anomaly involves, above all, the stomach and duodenum. Prior to the introduction of imaging techniques such as endoscopic ultrasonography, computer tomography and nuclear magnetic resonance, the diagnosis could only be made by means of an operative procedure. Treatment is primarily surgical; smaller cysts can be treated by endoscopic fenestration. Pharmacological treatment with somatostatin is rarely performed. PMID- 12635472 TI - Serratia marcescens in the neonatal intensive care unit: re-emphasis of the potentially devastating sequelae. AB - BACKGROUND: It is known that infections with Serratia marcescens can take a progressive course in preterm infants and that meningoencephalitis with this pathogen exhibits an extremely bad neurologic prognosis. METHODS AND RESULTS: We report on five cases of septicemia with Serratia marcescens in preterm infants during a nosocomial outbreak. Three patients developed meningoencephalitis with brain abscesses. Mild clinical and laboratory findings of infection contrasted with destructive findings on MRI scan. All five patients survived, those with isolated bacteremia without neurologic sequelae. CONCLUSION: When Serratia marcescens is isolated from any source in a neonatal intensive care unit, preventive measures including strict hygiene and cohorting of infants must be implemented immediately since this pathogen seems to exhibit specific affinity for the central nervous system and Serratia marcescens meningoencephalitis takes a progressive and destructive course despite antibiotic therapy. PMID- 12635473 TI - The economic burden of sepsis in Austria. PMID- 12635474 TI - [Role of changes in the transmembrane ion currents in pathological states of the body]. AB - Modern data of study of transmembrane ions currents (through the sodium-potassium and calcium-channels) in the mechanism of pathological pain syndromes were reviewed. Tetrodotoxyn-resistant sodium currents, abnormal combinations of different types of sodium-channels during the inflammation, neuropathies (syndromes hyperalgesii and allodonii) and diabetic neuropathy analyzed. The role of pain sensitivity transmitters was demonstrated. Nerve Grow Factor plays the important role in the sodium-channels genes expression. Different types of transmembrane calcium currents participated in hyperalgesium and hypoalgesium analyzed. The summary mechanism of pain suggested as voltage-calcium-channels activation and transsynaptic NMDA receptors excitation that should be conductive the neurons hyperexcitation and force of synaptic transmission. PMID- 12635475 TI - [New role of a highly-stable oligopeptides, neurotrophins, and immunomodulators in the regulatory continuum]. AB - We propose new approach to classification of regulatory peptides and proteins. It is based on the complex of their properties: stability, affinity to receptors, existence of specialized percursors, protein-transporters etc. In particular we describe very stable factors such as new peptide family named glyprolines. PMID- 12635476 TI - [Cholinergic mechanisms in the pathogenesis of genetically-caused absence epilepsy]. AB - Frontoparietal cortex and the thalamocortical circuit comprising reticular thalamic nucleus (RTN) and relay nuclei of the ventrolateral thalamus (VLT) are critical structures in the generation of spike-wave discharges (SWD) during absence seizures. The activity of these nuclei is under the control of the ascending cholinergic projections of nucleus basalis of Meynert. The aim of our study is to make an attempt to change the pattern of SWD in WAG/Rij rats by injecting of cholinotoxine AF64A to the area of RTN. Spontaneous SWD were registered in cortex of WAG/Rij rats with genetically determined absences. The spectral content of SWD was analyzed by means of the Fast Fourier Transformation (FFT) procedure. Unilateral injections of AF64A (1 nmol) to RTN led the decrease in duration and number of SWD comparing to the basal EEG recordings 2 days after the lesion. The FFT analysis showed the disappearance of 17-18 Hz spike on the side of the lesion compared with the intact side. The immunohistochemical study for acetylcholinetransferase (ChaT)-containing neurons showed the loss of ChaT positive cells in the nucleus basalis area on the side of the lesion. The removal of cholinergic afferentation of RTN and cortex from nucleus basalis inhibits the SWD developing most likely due to the decrease of cortical excitability. Moreover, possibly cholinergic transmission is involved in the transforation of the synchronized phenomena (SWD) to another with close mechanism of generation. PMID- 12635477 TI - [Features of regulating delta sleep-inducing peptide by free radical processes in tissue and erythrocyte membranes in intact animals and under stress]. AB - In activity the comparative analysis of metabolic effects delta--sleep inducing peptide (DSIP) in tissues and erythrocytes of intact rats and under cold stress is conducted. The regulation effect of DSIP in attitude of free radical processes will be realised through modulation the prooxidant--antioxidant balance: both for intact animal, and at stress. Exogenous DSIP increases the antioxidant system activity in tissues of brain, liver and blood in standard conditions and under cold stress. The anti-stress effect of DSIP is directed as on increase of power endogenic enzymatic antioxidant system activity, specially glutathione peroxidase activity, and not enzymatic of antioxidant protection. The DSIP renders different influence on activity of prooxidant enzymes: for intact animal boosts the myeloperoxidase activity in blood neutrophils, not rendering essential influencing on the xanthine oxidase activity in tissues of brain, liver and activates the myeloperoxidase activity, depresses the xanthine oxidase activity for rats at stress. The membranotropic effect of DSIP in the norm and under stress is connected to increase of stability of protein--lipid interplays. The membranostabilizing effect of DSIP in conditions of stress is characterized decrease of polarity of lipid phase and negative surface charge of erythrocyte membranes, modified in course of lipid peroxidation. PMID- 12635478 TI - [Genetic and immunologic aspects of type 1 diabetes mellitus]. AB - Prediction of type 1 diabetes mellitus (IDDM) and its identification in preclinical period is one of the central problems in modern medicine. They are based comprehensive genetic, immunologic and metabolic evaluations. We observed four hundred seven first-degree relatives of patients with IDDM (240 families in which one of the children or one of the parents had IDDM) have been included in the study. The study of HLA-DQA1, HLA-DQB1 polymorphic alleles and DRB1 genes and their combinations. The genetic study included searching HLA loci (HLA-DQA1, HLA DQB1 polymorphic alleles and DRB1 genes) loci. To evaluate the genetic risk two approaches we used: first--carrying predisposing HLA-DQ alleles and DRB1-genes and it's combination (mainly associated in Russian population was DRB1*04 DQB1*0302, DRB1*04-DQA1*0301, DQA1*0301-DQB1*0302, DQA1*0301-DQB1*0302 and four susceptible alleles in A- and B- chains (Asp 57-, Arg 52+)) and second--IBD (identity by descent), in Russian population HLA-identical for 2 haplotypes sibs had risk of development of IDDM of 18%, for 1 haplotype--3%, for 0 haplotype 0.9%. The antibodies (ICA, IAA) prevalence rate has not depended on availability of predisposing HLA-DQ alleles and DRB1-genes and haploidentity of normal sibs and sibs with IDDM. However, GADA prevalence rate in groups having high predisposed alleles has been noticed as significantly higher (28.6%) comparing with 7.7% in groups that had no predisposing alleles (p < 0.05). The comparison of antibodies prevalence rate to sibs HLA-identity has shown the significant increase or GADA prevalence rate in group of siblings identical for one haplotype comparing with non-identical sibs (27.3% and 0% respectively, p < 0.001). PMID- 12635480 TI - [Age-related involution of organs and tissues]. AB - Contemporary civilization confronts the humankind with the challenge of rapid ageing, which becomes especially relevant for developed countries. This necessitates the elaboration of optimal approaches to the management of ageing, which would enable a radical prolongation of active, full-value, working period of human life and, thus, would reduce the percentage of age-related disability. An essential role in the successful accomplishment of these objectives consists in the search for the mechanisms of age-related involution of organs and tissues. This overview presents and analysis of the modern concepts of these mechanisms. PMID- 12635479 TI - [Current trends on cholinergic effect on secretion and activity of renin and angiotensin-converting enzyme]. AB - This article concerned to insufficiently studied subject on regulatory influence of cholinergic system on key parameters of renin-angiotensin system (renin and angiotensin converting enzyme activity and secretion). Literature data and author's investigations are considered. Influence of systemic and intrarenal infusions of acetylcholine, its analogues and antagonists, inhibition of vagal activity, denervation of kidney, increased and decreased parasympathetic tonus on renin and angiotensin converting enzyme secretion is discussed. Molecular mechanisms of acetylcholine influence on juxtaglomerular cells are considered. Regulatory significance of cholinergic influence on reninangiotensin system is discussed. PMID- 12635481 TI - HMO mandates doom risk contract for cardio network. PMID- 12635482 TI - Shared medical appointments save money for capitated groups. AB - Shared medical appointments aren't money-makers for Palo Alto Medical Foundation, but they do cut expenses, improve patient healing and free up physician schedules. The group nature of the encounters also encourages patients to follow medical orders. Find out more. PMID- 12635483 TI - Preparation can take the sting out of catastrophic care costs. AB - Managing care of high-cost capitated patients can be a real challenge. But there are basic strategies capitated groups can use to minimize the costs before they reach catastrophic proportions and become budget busters. PMID- 12635484 TI - Only a fraction of doctors use the internet to save money. AB - Although three out of four physicians now use the Internet, a survey from the AMA suggests that only one of four connects with the web for such business functions as transferring medical records or obtaining managed care data, thus missing out on a potentially important source of savings and efficiency under capitation. PMID- 12635485 TI - [Stalking--a modern manifestation of erotomania?]. AB - Stalking is the technical term for the systematic, excessive and sometimes offensive molestation in order to seek personal contact with a target person, mostly of the contrary sex. This phenomenon seems to have increased recently and concerns above all celebrities of several kinds. In some cases, stalking behavior can culminate in acts of violence. However, the pathological fixation onto an often unattainable person is not a modern phenomenon, since it was studied and described under the term erotomania in former centuries. The author presents and discusses the case of a young man, who displayed his stalking behavior exclusively by telephone calls. PMID- 12635486 TI - [Weather-induced changes in cannabinoid content of hair]. AB - Authentic hair samples from Cannabis users and a drug free hair sample which was separately spiked with tetrahydrocannabinol (THC), cannabidiol (CBD) or cannabinol (CBN) were exposed outside as well as to natural sunlight at prevailing and elevated humidity in quartz glass tubes during 8 weeks. In addition, authentic and spiked hair samples were exposed to xenon arc radiation in a light exposure cabinet for 24 hours. Stability of THC, CBD and CBN in authentic samples differed from that of the spiked hair. The radiation experiment revealed that CBN could not be measured in hair which had been spiked with THC. Under all conditions chosen the concentrations of THC, CBD and CBN decreased. At high humidity the concentrations declined more rapidly. In both authentic and spiked samples THC was most unstable compared to CBD and CBN. Therefore, in hair analysis determination of CBD and CBN seems promising to detect Cannabis exposure even under unfavorable conditions. PMID- 12635487 TI - [Suicidal yew poisoning--from Caesar to today--or suicide instructions on the internet]. AB - Already the Celts and ancient Germanic peoples knew about the poisonousness of the yew, which played an important part in the mythology of these civilizations. For hunting, the arrows were made poisonous with yew juice, and yew leaves were used for homicide and suicide. In modern times taxine is rarely used with suicidal intent, although this method is actually recommended on the respective Websites. After a 14-year-old boy had intensively studied poisonous plants and methods of suicide on various Websites, he cut leaves from a yew tree (taxus baccata) in his parents' garden, crushed and ingested them and died soon afterwards. At the forensic autopsy pieces of the partially crushed, partially completely preserved yew leaves were found in the stomach. The histological findings were unspecific, e.g. marked general blood congestion of the internal organs and pronounced cerebral and pulmonary edema. When the tree leaves found in the stomach were viewed under the light microscope, a stoma typical of taxus was observed; chemical-toxicological investigations revealed 3,5-dimethoxyphenol in the gastric content, which is considered a marker for the ingestion of taxus. PMID- 12635488 TI - [An unusual area of injury in postmortem animal feeding beneath clothing]. AB - The authors report on a woman who died on the age of 75 years and who was found dead in her dwelling with an extensive injury of the left thigh two days after the last sign of life. Together with the corpse a poodle was present in the dwelling. Due to the unusual localization totally beneath the clothes (trousers) and the morphology of the injury an unnatural cause of death was suspected. According to the autopsy findings and the histological results, postmortal animal scavenging caused by the poodle remained the only explanation for the injury. Supplementary DNA testing yielded final certainty. PMID- 12635490 TI - [Possible applications of Y chromosome STRs in examination of abortion tissue]. AB - By application of Y-chromosomal STRs, DNA analysis of abortion material can be considerably facilitated since great excess of maternal DNA is tolerated without disturbing the Y-STR amplification. If paternity can't be excluded on the basis of the Y-STR haplotype, further examinations must follow, e.g. autosomal STR analysis. For this purpose, histological preparation of the abortion tissue might still be necessary. Different Y-chromosomal haplotypes of embryo and putative father usually lead to an exclusion from paternity. Based on four case examples, the feasibility of this method is discussed. PMID- 12635489 TI - [An unusual homicide case with subsequent suicide]. AB - In the medicolegal literature reports on homicides followed by suicide, especially in elderly people, are rare. In the present case, the victim, a 79 year-old woman, showed a rather unusual injury pattern: 3 gunshots to the head (with only 2 entrance wounds and 3 intracranial projectiles) and 2 stabs from a knife. The tools used for inflicting the stab wounds (a hunting knife and a double-edged dagger) were left in the victim's body after the offense. The 89 year-old husband hanged himself with an aerial cable after killing his wife. The police investigations and the autopsy findings suggested a combined homicide suicide. The motive for the offense could be a so-called delusion of poverty associated with symptoms of depression. PMID- 12635491 TI - Molecular, cellular and medical aspects of the action of nutraceuticals and small molecules therapeutics: from chemoprevention to new drug development. AB - Dietary supplements, functional foods and their concentrated, sometimes purified, active forms, the so-called nutraceuticals, are becoming increasingly popular throughout the world. Small molecules that regulate signal transduction cascades and gene expression are being tested by many pharmaceutical companies. A rapidly and exponentially growing industry (close to $30 billion in 1999 worldwide) exists to commercialize and exploit this interest. However, the scientific basis of the action of such unproved products is in the very early stages of development. While supporters claim they produce miracle cures, opponents argue that such unproved agents do more harm than good. PMID- 12635492 TI - Effect of D-003, a mixture of high molecular weight primary acids from sugar cane wax, on CL4C-induced liver acute injury in rats. AB - D-003 is a mixture of very high molecular weight aliphatic primary acids purified from sugar cane (Saccharum officinarum, L.) wax, in which octacosanoic acid is the most abundant component. Previous experimental studies have shown that D-003 not only shows cholesterol-lowering and antiplatelet effects, but also offers strong protection against plasma lipoprotein oxidation. Acute hepatotoxicity induced by CCL4 in rats has been related to an increased rate of lipid peroxidation, and different antioxidant compounds have been revealed to be effective in this model. The aim of this study was to investigate the effects of D-003 in acute hepatotoxicity induced by CCL4 in rats. Male Sprague Dawley rats were randomly distributed in four experimental groups as follows: group 1: negative control rats; group 2: positive control rats (CCL4-treated); groups 3 and 4 rats with liver damage induced by CCL4 and treated with D-003 at 25 and 100 mg/kg, respectively. Acute liver injury was induced by CCL4 suspended in olive oil and intraperitoneally administered at 1 ml/kg. Eighteen hours after CCL4 dosing, the rats were anesthetized with ether and their livers were removed for histopathological studies. D-003 at 25 and 100 mg/kg significantly (p < 0.01) decreased the percentage of ballooned cells and hepatocytes with lipidic inclusions and increased the percentage of normal hepatocytes compared with that in positive controls in a dose-dependent manner. The percent inhibitions of the occurrence of ballooned cells and hepatocytes with lipids were marked (75% and 50%, respectively) with the high dose (100 mg/kg). The percent of turgent hepatocytes was also significantly reduced compared with that in positive controls, but this effect was not dose-dependent. No histological alterations in the liver sections of negative controls were found. Necrotic areas and inflammatory infiltrate were observed in the liver of 7/8 (87.5%) of positive controls. In turn, D-003 dramatically reduced both necrotic areas and inflammatory infiltrate and was present in only 1/8 (12.5%) animals treated with D-003 25 mg/kg and in none (0%) of the animals treated with 100 mg/kg. D-003 protected against the histological changes characteristic of CCL4-induced hepatic injury in rats, in which the process of lipid peroxidation plays the main role. The relationship between this protective action of D-003 on this experimental model and its antioxidant effects needs to be further investigated before definitive conclusions are drawn. PMID- 12635493 TI - Safety of stabilized, orally absorbable, reduced nicotinamide adenine dinucleotide (NADH): a 26-week oral tablet administration of ENADA/NADH for chronic toxicity study in rats. AB - The safety of the stabilized, orally absorbable form of reduced nicotinamide adenine dinucleotide (NADH), known under the brand name ENADA, was investigated over a period of 26 weeks. Eighty healthy rats (40 males and 40 females) were divided into two groups. One tablet ENADA/NADH 5 mg per day was administered orally to one group while identical-looking white tablets not containing NADH (placebo) were given to the other group. The following parameters were statistically analyzed: body weight, body weight gain, food consumption, hematology, clinical chemistry, organ weight and organ histology. Clinical signs and mortality were recorded. There were no deaths associated with the study drug and no treatment-releated clinical signs. No differences in body weight between the placebo and the ENADA-treated males were observed. In the second half of the treatment period (weeks 13-26) females treated with NADH gained significantly (p < 0.05) more body weight than the controls. Food consumption in the treated males was similar to that in controls. From approximately week 15, the treated females consumed up to 10% more food than the controls. No differences were observed between the control and the treated groups in terms of hematology or clinical chemistry parameters. There was no apparent treatment-related effect on urine analysis parameters or on either the absolute or the relative organ weight. Furthermore, no macroscopic evidence of specific target organ toxicity associated with the test drug was observed. Histological findings in the treated rats were generally similar to those in control rats. A daily dose of 5 mg in a rat corresponds to a dose of 175 mg per day in a 70-kg human. This is 175 times the recommended daily dosage of 1 ENADA tablet per day. Hence ENADA/NADH 5 mg tablets can be generally regarded as safe. PMID- 12635494 TI - Seijo-bofu-to, Jumi-haidoku-to and Toki-shakuyaku-san suppress rashes and incidental symptoms in acne patients. AB - Seijyo-bofu-to, Jumi-haidoku-to and Toki-shakuyaku-san effectively suppressed acne rashes as well as incidental symptoms. The synergistic activities of the ingredients in the Kampo formulations might produce these effects. In contrast, distinct suppression of incidental symptoms was not found with antimicrobials. The cause of adverse effects in antimicrobials has not yet been clarified and different degrees of suppression of incidental symptoms among the Kampo formulations exist. PMID- 12635495 TI - KE-758, an active metabolite of the new anti-rheumatic drug KE-298, suppresses production of tumor necrosis factor-alpha and interleukin-1 beta in THP-1, a human monocyte cell line. AB - KE-758, an active metabolite of KE-298, is a novel sulfhydryl antirheumatic drug. We analyzed the effect of KE-758 on tumor necrosis factor (TNF)-alpha and interleukin (IL)-1 beta production by a human monocytes cell line (THP-1 cells), stimulated with interferon-gamma (IFN-gamma) plus lipopolysaccharide (LPS). We compared the effects with other thiol-modifying antirheumatic drugs such as D penicillamine, bucillamine and auranofin. THP-1 cells were treated with IFN-gamma for 16 h and were then exposed to LPS for an additional 6 h (for TNF-alpha detection) or 24 h (for IL-1 beta detection). The amounts of TNF-alpha and IL-1 beta in culture supernatants were measured using enzyme-linked immunosorbent assay. KE-758 and auranofin but not D-penicillamine and bucillamine significantly suppressed both TNF-alpha and IL-1 beta. Auranofin suppressed IL-1 beta production by reducing cellular viability. Reverse transcriptase-polymerase chain reaction analysis revealed that the suppressive effect of KE-758 is based on the inhibition of messenger ribonucleic acid expression of TNF-alpha and IL-1 beta. KE-758 had no effect on p75 and p55 soluble TNF receptor production in IFN-gamma and LPS-stimulated THP-1 cells. Thus, KE-758 inhibits both TNF-alpha and IL-1 beta production and its antirheumatic profile is apparently distinct from that of D-penicillamine, bucillamine and auranofin. PMID- 12635496 TI - Pharmacokinetics of oxatomide in preterm infants. AB - The pharmacokinetics and tolerability of oxatomide oral suspension were investigated in preterm infants to evaluate the feasibility of planning a further study to assess its antiinflammatory effects and its effectiveness in preventing chronic lung disease (CLD). Following the administration of oxatomide 1 mg/kg, the peak plasma concentration (Cmax), the elimination half-life (t1/2), the volume of distribution (Vd), and the area under the curve (AUC) 0-36 h were measured and the following results were obtained: 42.2 +/- 15 ng/ml at 2 h after oxatomide administration, 41.4 +/- 2.0 h, 37.4 +/- 4.2 l/kg, and 468 +/- 52 ng/ml/h, respectively. Our study, therefore, demonstrated that a dose of 1 mg/kg/day oxatomide was effective in reaching therapeutic plasma levels in preterm infants without inducing adverse effects. PMID- 12635497 TI - Applications of polymerase chain reaction in rheumatology. AB - Polymerase chain reaction (PCR) is a highly sensitive and specific method for detection and quantification of specific nucleic acids from a clinical sample. With its use, genetic, infectious, neoplastic, and autoimmune diseases can be diagnosed and managed with a high level of sensitivity, accuracy, and rapidity. This technique exactly reproduces unlimited copies of DNA, even if only a small amount are present initially. PCR assays can detect presence of fastidious and slow-growing microorganisms, such as chlamydia, mycoplasmas, mycobacterias, and viruses directly from clinical specimens and also can detect antimicrobial resistance. The value of viral load measurement by nucleic acid amplification in the management of patients with HIV infection or hepatitis C has also been well established. From the point of view of a clinician, the applications of PCR are focused mainly in the amplification and detection of diagnostic DNA segments from the genomes of both pathogens and patients. PMID- 12635498 TI - Role of bacteria and HLA-B27 in the pathogenesis of reactive arthritis. AB - Strictly speaking, "reactive arthritis" is a conventional term with no study verified definition. This review will focus on the type of arthritis that is induced by the following species: Chlamydia, Shigella, Salmonella, Yersinia, and Campylobacter. The types of arthritis caused by these pathogens share a clinical pattern that is common in the spondyloarthropathies, especially undifferentiated spondyloarthropathy and Reiter's syndrome. All these diseases, including ankylosing spondylitis, must also share major pathogenetic pathways. PMID- 12635499 TI - Reactive arthritis: newer developments. AB - Reactive arthritis (ReA) is characterized by an aseptic inflammatory articular involvement occurring in a genetically predisposed individual secondary to an infectious process localized outside the joint. ReA usually refers to an acute or insidious oligoarthritis process after enteric (enteroarthritis) or urogenital (uroarthritis) infection. Conventional antirheumatic therapeutic modalities based on nonsteroid anti-inflammatory drugs, sulfasalazine, and steroids are effective in the majority of patients. In more refractory cases, the use of second-line agents including methotrexate and more recently biological agents such as etanercept and infliximab has been found highly effective. The role of antibiotics remains not well established, although they appear to be effective in acute ReA of urogenital origin. PMID- 12635500 TI - Advances in the management of septic arthritis. AB - Bacterial arthritis still is a common and serious problem at mayor urban medical centers and is one of the most rapidly destructive forms of acute arthritis. The yearly incidence of bacterial arthritis varies from 2 to 10 per 100,000 in the general population to 30 to 70 per 100,000 in patients with rheumatoid arthritis and in patients with joint prostheses. Irreversible loss of joint function may develop in up to 50% of the patients. Despite better antimicrobial agents and improved hospital care, the fatality rate for this medical problem has not changed substantially during the past 25 years. An understanding of the risk factors and the pathogenesis of nongonoccocal bacterial arthritis and other forms of infectious arthritis, primarily in the context of a differential diagnosis and treatment, are important in order to avoid the delay in making a correct diagnosis and to improve the prognosis. PMID- 12635501 TI - Soft tissue disease. AB - In the assessment of patients with soft tissue complaints, it is important to consider infectious etiologies in the differential diagnosis, especially in immunocompromised hosts. The exact categorization of some bacterial infections of the soft tissues may be difficult. The structures potentially involved include the skin, subcutaneous tissue, fascia, and skeletal muscle. Classification is usually based upon the anatomic structure involved, the infecting organism, and the clinical picture. The categorization is complicated by the fact that some infections may involve several soft tissue components and multiple bacterial species. In this review, we will cover cutaneous and subcutaneous tissue infections, fasciitis, septic bursitis, tendonitis, and pyomyositis. PMID- 12635502 TI - Imaging of osteomyelitis and musculoskeletal soft tissue infections: current concepts. AB - The diagnostic imaging of osteomyelitis can require the confluence of multiple imaging technologies. Conventional radiography should always be the first imaging modality. Sonography is most useful in the diagnosis of fluid collections in a joint or in the extra-articular soft tissues but is not useful for evaluating presence of osseous infection. CT scan can be a useful method to detect early osseous erosion and to document the presence of sequestrum, foreign body, or gas formation but generally is less sensitive than other modalities for the detection of bone infection. Nuclear medicine and MRI are the most sensitive and most specific imaging modalities for the detection of osteomyelitis. Nuclear medicine is particularly useful in identifying multifocal involvement, which is common in children. MRI provides more accurate information of the local extent of the soft tissues and possible soft tissue abscess in patients with musculoskeletal infection. PMID- 12635504 TI - Role of endogenous retroviruses in autoimmune diseases. AB - Endogenous retroviruses (ERVs) correspond to the integrated proviral form of infectious retroviruses that are trapped within the genome by mutations. Endogenous retroviruses represent a key molecular link between the host genome and infectious viral particles. Proteins encoded by ERVs are recognized by antiviral immune responses and become targets of autoreactivity. Activation of ERVs, such as human ERV-K or a human T-cell lymphotropic virus-related endogenous sequence, may also mediate pathogenicity of Epstein-Barr virus. Endogenous retrovirus peptides can directly regulate immune responses. Thus, molecular mimicry and immunomodulation by ERVs may account for self-reactivity and abnormal T- and B-cell functions in autoimmune disorders. PMID- 12635503 TI - Hepatitis C virus and arthritis. AB - Arthritis is one of the several autoimmune disorders induced by HCV infection. There is not a specific clinical pattern of HCV-related arthritis, but two nonerosive subsets have more frequently been described: a RA-like polyarthritis and a less common mono-oligoarthritis involving medium-sized and large joints, often showing an intermittent course. This latter form is associated with the presence of serum cryoglobulins. Because of its variable characteristics, HCV related arthritis must be considered in the differential diagnosis of many patients having inflammatory joint involvement. Antikeratin antibodies and possibly IgA RF can be useful in distinguishing between RA and HCV-related RA like polyarthritis. In fact, these tests are highly specific in RA patients. In any case, the search for HCV antibodies should be more widely performed in the diagnostic approach to rheumatic diseases. An association between PsA and HCV infection has been described in the literature, but the authors were unable to confirm these data. Nonsteroidal anti-inflammatory drugs, hydroxychloroquine, and low doses of corticosteroids are the cornerstones of the treatment of HCV-related arthritis. An etiologic therapy with alpha-interferon and ribavirin is useful when required by hepatic or systemic involvement; such therapy could also be considered in selected cases of isolated arthritis that are unresponsive to other drugs. Few case reports described the onset of polyarthritis after the administration of alpha-interferon for HCV-related chronic hepatitis. This topic should be more accurately studied in the future to exclude a spurious association between the two events. PMID- 12635505 TI - Rheumatic manifestations of human immunodeficiency virus infection. AB - Rheumatic complaints are common in patients with human immunodeficiency virus (HIV) infection. With the advent of the modern combined antiretroviral treatment, life-long control of HIV infection and normalization of life expectancy in HIV positive patients have become realistic perspectives, but new rheumatic complications, such as osteoporosis, osteonecrosis, gout, and mycobacterial and mycotic osteoarticular infections may be more prevalent. Rheumatologists, internists, and general physicians need to be familiar with the presentation and treatment of these conditions in HIV-positive patients. PMID- 12635506 TI - Infections in systemic connective tissue diseases: systemic lupus erythematosus, scleroderma, and polymyositis/dermatomyositis. AB - In SLE, scleroderma, and PM/DM, infections are important causes of morbidity and mortality. This increased risk of developing infections is the result of immune abnormalities and of organ system manifestations associated with these diseases and their treatments. Common bacteria are responsible for most mild and lethal infections; however, opportunistic microorganisms cause death in some patients, particularly in those receiving high doses of corticosteroid and immunosuppressive therapy. Various viral and fungal infections also contribute to the morbidity and mortality associated with these diseases. Regardless of the cause of infections, adequate and prompt recognition and proper treatment of the infected patient are imperative. Thus, patients with these diseases, especially when receiving high doses of corticosteroids and immunosuppressive therapy, need to be monitored closely for these infections. This care and concern is necessary to ensure optimal patient outcomes, both in terms of morbidity and mortality. PMID- 12635507 TI - Infection complications associated with the use of biologic agents. AB - Despite the considerable clinical, radiologic, and functional benefits of biologic inhibitors in inflammatory arthritides, some concern exists regarding the occurrence of infections in patients treated with these agents. Clearly, comorbidities such as diabetes mellitus, heart disease, disability, and concurrent immunosuppressive medication all contribute to the risk of infection. Increased and closer observation may be in part responsible for some of the reported increases in the rates of mild infections with these drugs. The development of serious infections, particularly TB, in patients taking infliximab seems to be greater than would be expected in this population. Furthermore, experimental data from in vitro investigations and animal models demonstrate a link between decreased TNF alpha activity and increased susceptibility to TB. Why some patients, but not others, succumb to rapidly disseminated infection is unknown but may be related to the extent of TNF inhibition in different individuals. This difference in inhibition may also explain why the incidence of TB seems to be increased with infliximab in comparison with the other TNF blockers. Attribution analysis is the method used to assess the likelihood of a connection between two occurrences and includes such factors as temporal association, few alternative explanations, analogy with similar cases, and biologic plausibility. The putative relationship between anti-TNF treatment and infection is further strengthened by the presence of these factors [101]. Continued vigilance is therefore required in the use of biologic agents in patients with RA, most of whom are already in some way immunocompromised. Everyone who is under consideration for such treatment should be carefully evaluated for the presence of infection, and prophylactic antituberculous therapy should be used if latent TB is discovered. Both patients and primary physicians need to be aware of the possibility that serious infection may develop; if such a problem is diagnosed, the biologic inhibitor should be discontinued until adequate treatment has been completed. Caution is advised in patients with recurring infections and in those with severe comorbidities, for example, poorly controlled diabetes mellitus or heart failure. Administration of live vaccines to patients taking these drugs is not recommended, but patients should be brought up to-date with all immunizations relevant to their age group before commencement of therapy. Physicians prescribing biologic agents should be encouraged to report any suspected drug-related adverse event. Long-term observation will be required to determine the exact nature of the relationship between cytokine inhibition and infection. PMID- 12635508 TI - Therapeutic extradural cortical stimulation for movement disorders: a review. AB - Extradural motor cortex stimulation was introduced in 1989 for control of central pain. In recent years this has been found useful in several patients with movement disorders. This paper attempts to bring together all the relevant literature, discuss mechanisms and lay out guidelines for future research and clinical applications. PMID- 12635509 TI - Chronic motor cortex stimulation for movement disorders: a promising perspective. AB - Five patients affected by thalamic hand and international myoclonus have been evaluated and selected for chronic motor cortex stimulation. A quadripolar electrode strip was placed epidurally under local anesthesia through an MR-image guided single precentral burr hole placed following the morphologic recognition of the central sulcus. Intra-operative stimulation was used to induce muscle contraction at the affected site to confirm the correct placement of the electrode. A one-week trial period preceded the implant of an internal pace-maker under general anesthesia. A remarkable decrease in pain was reported by four patients together with the reduction of dystonia and rigidity in thalamic hand and marked decrease of intentional myoclonus. No complications or undesired side effects of electrode implant and stimulation were observed. PMID- 12635510 TI - The deafferented nonhuman primate is not a reliable model of intractable pain. AB - Before extending the application of motor cortex stimulation it is important to investigate the intimate mechanisms by which it alleviates intractable pain and to consider possible side effects. Self-mutilation in animals following extensive neurectomy or posterior rhizotomy of a limb is thought to reveal severe dysesthesias in the deafferented zone suggesting its usefulness as an animal model of chronic pain in humans. We here show in deafferented nonhuman primates that the autotomy behavior immediately follows the surgery and disappears after 28 days. In keeping with the experience of Y. Lamarre, the simple but careful care of all wounds is sufficient to abolish this behavior. Our results do not exclude the possibility that the deafferentiation is still painful for the monkeys, but they definitely rule out that autotomy is a consistent response to deafferentation. PMID- 12635512 TI - Corticospinal volleys evoked by transcranial stimulation of the brain in conscious humans. AB - The direct recording in conscious humans of corticospinal volleys evoked by different magnetic and electric techniques of transcranial stimulation demonstrates that it is possible to activate neurones of the motor cortex in several different ways. Lateral electrical stimulation of the motor cortex preferentially activates the axons of corticospinal neurones in the subcortical white matter, and evokes a D-wave in pyramidal tract. The way of activation of corticospinal neurones using magnetic stimulation depends on the direction of the electrical current induced in the brain and on the shape of the coil. Monophasic magnetic stimulation with a focal figure-of-eight coil inducing posterior anterior current in the brain activates corticospinal neurones trans-synaptically recruiting an 11-wave, with later I-waves appearing in sequence at higher intensities and a D-wave at very high intensities. If the induced current is rotated to the anterior-posterior direction late I-waves are preferentially recruited and when a D-wave is recruited, it has a later onset than the electrical D-wave, suggesting an activation nearer the cell body of the pyramidal neurones. A latero-medial induced current activates both corticospinal axons at the same point as electrical stimulation evoking a D wave and cortico-cortical axons evoking I-waves. A nonfocal large circular coil centered at the vertex is capable of activating pyramidal neurones both at the initial segment and trans synaptically evoking a D wave with a longer latency than the electrical D-wave and I-waves. Using a biphasic magnetic stimulation, both phases of the biphasic pulse are capable of activating descending motor output and the pattern of recruitment of descending activity depends on the intensity of the stimulus and the relative threshold of each volley to each direction of current flow. PMID- 12635511 TI - Pain-related modulation of the human motor cortex. AB - Pain is a complex multi-dimensional phenomenon that influences a wide variety of nervous system functions, including sensory--discriminative, affective- motivational and cognitive--evaluative components. So far, these components have been studied in both patients with chronic pain and in normal subjects in whom pain was induced experimentally. The interaction between pain and motor function is not fully understood, although from everyday life it is known that pain affects movements. The effects of pain on motor control are typically seen as a limited or impaired ability to perform movements. Most studies have dealt with the effects of pain on the spinal cord reflexes, but in recent years, several lines of evidence suggest that the interaction between motor and pain systems in conditions of pain induced experimentally, rather than a simple spinal reflex, is a more complex process that involves also supraspinal brain areas. Although pain motor interaction shows different features and time course depending on different pain variables, such as duration (tonic versus phasic pain), submodalities (deep versus superficial pain) and location (distal versus proximal pain), a common finding is that pain is able to inhibit the motor cortex. This motor cortex inhibition may act as a sort of motor 'decerebration' so as to allow the spinal motor system to freely develop protective responses to noxious stimulation. Further studies are required to assess the effects of pain on the motor system in patients suffering from chronic pain, in order to develop innovative rational therapeutic strategies to reduce both pain and motor disability. PMID- 12635513 TI - Low-rate repetitive TMS allays central pain. AB - Only about 50% of central pain patients respond to motor cortex stimulation in the long run. There is a need for prognostic factors. Here we show that propofol test and TMS both predict short-term effect in nine patients with central pain. This may help reduce the number of failures. PMID- 12635514 TI - Motor cortex stimulation in patients with post-stroke pain: conscious somatosensory response and pain control. AB - We analyzed the conscious sensory responses to cortical stimulation of 31 patients with post-stroke pain who underwent motor cortex stimulation (MCS) therapy. During surgery for electrode placement, a sensory response (tingle projected to a localized peripheral area) was elicited by high-frequency stimulation (50 Hz) in 23 (84%) from the somatosensory cortex, and in 16 (52%) from the motor cortex without muscle contraction. Unpleasant painful sensation was induced or their original pain was exacerbated in 12 patients (39%) when the somatosensory cortex was stimulated and in two (6%) when the motor cortex was stimulated. Somatosensory responses were induced in eight (25%) even by low frequency stimulation (1-2 Hz) of the motor cortex at an intensity below the threshold for muscle contraction. In contrast, among 20 nonpain patients who underwent a similar procedure for cortical mapping in epilepsy or brain tumor surgery, a sensory response was produced by high-frequency stimulation in only eight (40%; p < 0.02) from the somatosensory cortex and four (20%; p < 0.03) from the motor cortex. Pain sensation was not induced by stimulation of the somatosensory cortex (p < 0.002) or motor cortex in any of these patients. In addition, none of these patients reported a sensory response to low-frequency stimulation. In both of the two post-stroke pain patients who reported abnormal pain sensation in response to stimulation of the motor cortex, MCS failed to control their post-stroke pain. These findings imply that the sensitivity of the perceptual system even to activity of the motor cortex is heightened in post stroke pain patients, which can sometimes hinder pain control by MCS. PMID- 12635515 TI - Motor cortex stimulation for neuropathic facial pain. AB - Facial neuralgia is the last common pathway for a variety of pathological conditions with different etiology. Neuropathic facial pain is often refractory to routine medical or surgical treatments. We present here a long-term follow-up of two patients with unilateral facial neuropathic pain due to idiopathic trigeminal neuropathy or to surgical trauma to the glossopharyngeal nerve, respectively. These patients have been treated by other modalities for several years without obtaining satisfactory pain relief. Electrical stimulation of the motor cortex (MCS) with a quadripolar electrode contralateral to the painful area of the face was attempted in both cases for control of the facial pain, and resulted in immediate analgesia with more than 50% pain reduction. During a follow-up period of 72 months, a sufficient (> 50%) and stable analgesic effect of MCS was observed. These cases are discussed and the recent literature on MCS is reviewed in an attempt to identify indications for MCS as well as key structures in the brain for mediating the MCS effect. PMID- 12635516 TI - Carotid stent angioplasty: the role of cerebral protection devices. AB - Carotid endarterectomy has been validated with results of several randomized controlled trials in which its effectiveness has been demonstrated over that of the best nonsurgical therapy. However, in the past several years, carotid angioplasty with stent placement has emerged as a potential safe and effective alternative to carotid endarterectomy. In this article we examine the current status of carotid angioplasty with the recent introduction of innovative cerebral protection devices and improved endovascular devices. We present a brief description of the current randomized trials evaluating carotid endarterectomy compared to carotid angioplasty as well as our combined experience in 262 patients. PMID- 12635517 TI - Covered stent as an innovative tool for tumor devascularization and endovascular arterial reconstruction. AB - A temporal paraganglioma was pre-operatively embolised with particles and due to the fact that the internal carotid artery was encased by the tumor and that multiple petrous feeders precluded safe embolization, we decided to implant a balloon-expandable covered stent. On immediate post-procedural angiography, the arterial diameter was restored and a complete devascularization of the tumor was achieved. The tumor was operated with minimal blood loss and the carotid artery was released from surrounding tumor using the frame of the stent as a surgical limit. Covered stents may become a valuable tool for pre-operative tumor devascularization and arterial reconstruction especially when dealing with hypervascular tumors of the cranial base that encase the carotid artery. PMID- 12635518 TI - Incidental stereotactic diagnosis of cerebral insults. AB - Among the patients (6854 patients 1990-1999) who underwent computer-assisted stereotactic biopsy most were referred with the presumptive diagnosis of a brain mass lesion. Forty-three cases (0.63%) were found in which the final histopathological diagnosis excluded a neoplastic, infectious or inflammatory lesion but disclosed a cerebral insult. Histologically these could be subdivided into ischemic insults in 38 cases (88%) and hemorrhagic insults in five cases (12%). On the basis of clinical and radiological findings in this group, 35 patients (81%) were sent to our department because of suspected neoplasmatic lesions, two patients (5%) because of multiple sclerosis, two patients (5%) because of inflammatory disease and one patient (2%) because of a suspected infectious parasitic disease. All patients underwent initial CT examinations which showed hypodense lesions of the brain in 38 patients (88%) and hyperdense lesions in five cases (12%). Constant enhancement on CT scans of the mass lesion was found in 12 patients (28%) only. Fourteen lesions (33%) were located in the right hemisphere, five lesions (12%) in the left hemisphere, nine lesions (21%) in the basal ganglia, four lesions (9%) in the midbrain, two lesions (4.5%) in the corpus callosum and one lesion (2%) in a thalamus. Multiple lesions were present in eight cases (19%). The most common initial neurological symptoms upon clinical presentation were hemiparesis (18 patients, 42%), epilepsy (eight patients, 18%), a change in mental status (six patients, 14%). There was no mortality and no operative morbidity associated with the stereotactic biopsy in this group of patients. The most common neurological disorder, cerebrovascular insult, rarely poses diagnostic problems. If there are doubts a serial stereotactic biopsy can safely clarify the situation. PMID- 12635519 TI - Decreased nigral neuromelanin in Alzheimer's disease. AB - Using manual morphometric techniques, we estimated the amount of neuromelanin in hematoxylin and eosin-stained sections of the pars compacta of the substantia nigra of 19 Alzheimer's patients without nigral Lewy bodies and 12 age-matched controls. Our estimates showed that the mean area and areal fraction of neuromelanin were lower in Alzheimer's disease than controls but the number and size of the neuronal cell bodies, nuclei and nucleoli did not differ between the two groups. We speculated that the decreased amount of neuromelanin in nigral neuronal cell bodies could have resulted from neurofibrillary degeneration, retrograde degeneration from damage of nigral dopaminergic terminals in the striatum by the beta amyloid protein of the diffuse plaques and possibly transneuronal degeneration from damage of cell bodies or dendrites of nigral neurons by their plaque- and tangle-ravaged striatal, neocortical and other subcortical nigral connections. We hypothesized that any or all of the above types of degeneration could have lowered the rate of dopamine metabolism and the formation of one of its by-products, neuromelanin. Our study shows that a decrease in the amount of histopathologically-observable nigral neuromelanin commonly occurs in Alzheimer's disease without nigral Lewy bodies. PMID- 12635520 TI - Noninvasive assessment of cerebral oxygenation during high altitude trekking in the Nepal Himalayas (2850-5600 m). AB - Mountain trekking is significantly increasing in popularity. Hypoxia seems to play a key role in the pathogenesis of acute mountain sickness (AMS). The purpose of this study was to investigate regional cerebral (rSO2) and peripheral (SaO2) oxygen saturation for the first time, during 22 days high altitude trekking (measurement points: 3450, 4450, 4750, 5050 and 2850 m) in the Khumbu region of Nepal with near infrared spectroscopy and pulse oximetry. We examined 17 healthy volunteers 19-65 years old (8 female, 9 male; mean age +/- SD, 46.1 +/- 13.1 years). RSO2 and SaO2 were significantly (p < 0.001, ANOVA, Tukey test) decreased at high altitudes (4450, 4750 and 5050 m). The decrease in cerebral oxygen saturation was more pronounced at higher altitudes than in the periphery (rSO2/SaO2 = 0.56 at 5050 m). At higher altitudes (> 4450 m), two subjects showed reversible symptoms of AMS. The present data indicates that acute reduction in rSO2 values might be a primary cause of AMS, however, further studies and analysis are necessary to correlate our findings with cerebral symptom scores. PMID- 12635521 TI - Synaptic plasticity in thalamic nuclei enhanced by motor skill training in rat with transient middle cerebral artery occlusion. AB - The goal of this study was to determine if synaptic plasticity in the thalamus of rats subjected to stroke could be altered by motor training. Transient occlusion of right middle cerebral artery in adult female Sprague-Dawley rats (n = 35) was induced with an intraluminal filament followed by three training conditions, 1. motor skill training on Rota-rod requiring balance and coordination skills, 2. simple exercise on treadmill, and 3. nontrained controls. Synaptic plasticity in brain was evaluated by synapotophysin immunocytochemistry at 14 or 28 days after training procedures. Infarct volume was determined in Nissl stained sections. Both at 14 and 28 days after Rota-rod training, intense synaptophysin immunoreactivity was present in the right but not the left mediodorsal and ventromedial nuclei of thalamus of ischemic rats. In treadmill-trained animals, however, similarly intense synaptic plasticity in these two thalamic nuclei was seen only at 28 days. Immunostaining was found also in other brain regions adjacent to or remote from infarct site. The data suggest that motor training, particularly motor skill training involving balance and coordination, facilitates a uniquely lateralized synaptogenesis in the thalamus. PMID- 12635523 TI - Glutamate receptor antagonists modulate heat shock protein response in focal brain ischemia. AB - Neurons and glia reacting to ischemic injury exhibit delayed expression of heat shock proteins (HSPs). We tested the hypothesis that glutamate receptor antagonists alter neuronal and glial activation during focal cerebral ischemia, as shown by spatio-temporal changes in HSP immunoreactivity. Rats underwent focal ischemia by permanent occlusion of the middle cerebral artery. All animals were pre-treated with NBQX (30 mg kg-1), a competitive antagonist of the AMPA/kainate receptor, or CGS-19755 (10 mg kg-1), a competitive NMDA receptor antagonist, and euthanatized after 6 or 24 h of ischemia to demonstrate regional immunoreactivity of HSP-72 or 32 in brain. Neurons immunolabeled for HSP-72 appeared in the penumbral region adjacent to the infarct at 24 h and increased in number and distribution after pretreatment with NBQX or CGS-19755. Immunolabeling for HSP-32 revealed that pre-treatment with CGS-19755 caused ramified glia to infiltrate the ischemic cortex at 6 h, a pattern that was not seen in ischemic controls until 24 h. Blockade of the NMDA or AMPA/kainate receptor modulates cellular stress responses in both neurons and glia within the developing infarct. We conclude that early, rather than delayed, expression of HSP-32 is a sensitive indicator of glial activation induced specifically by CGS-19755. PMID- 12635522 TI - Glial cell line-derived neurotrophic factor protein prevents motor neuron loss of transgenic model mice for amyotrophic lateral sclerosis. AB - Effects of glial cell line-derived neurotrophic factor (GDNF) were studied in transgenic (Tg) mice model for amyotrophic lateral sclerosis. GDNF protein or vehicle was injected three times a week from 35 weeks of age into the right gastrocnemius muscle of Tg mice carrying mutant human Cu/Zn superoxide dismutase gene, and histological analysis was performed at 46 weeks. Clinical data showed a tendency of improvement, but was not significantly different between the two animal groups. In contrast, total number of and phospho-Akt (p-Akt) positive large motor neurons in the treated side was significantly more preserved in GDNF treated group than in vehicle group (p < 0.05). Immunoreactivity of phospho-ERK and active caspases-3 and -9 showed no difference. These results indicate that the intramuscular injection of GDNF protein prevented motor neuron loss while preserving survival p-Akt signal and without affecting caspase activations, suggesting a future possibility for the therapy of the disease. PMID- 12635524 TI - Multinevritis of cranial nerves following inhalation of toxins. AB - Cranial multineuropathy is an uncommon occurrence. We observed two cases of multinevritis of the cranial nerves which had many features in common: acute onset after toxic inhalation, occurrence of neuroparaxic block, and rapid resolution after corticosteroid therapy. We believe physicians should be aware that multinevritis of the cranial nerves may be related to acute exposure to toxic substances. Missing this diagnosis could lead to a delay in therapy. PMID- 12635525 TI - Alterations of cullin-5 mRNA levels in the rat central nervous system following hemorrhagic shock. AB - Hemorrhagic shock is a clinical syndrome that manifests as hypoperfusion, hypoxia, and ischemia initiating various cellular stress responses involved in the synthesis and release of an assortment of pro-inflammatory molecules, cytokines, chemokines, and reactive oxidant species (ROS). The ROS have been shown to oxidize and damage proteins making them targets for ubiquitination and proteasomal degradation. Cullin-5 (cul-5), an E3 ligase that binds ubiquitin to proteins targeted for degradation via the proteasome, was investigated for its gene expression during hemorrhagic shock. Male Long-Evans rats were subjected to volume controlled (27 ml kg-1) hemorrhage over 10 min and kept in shock for 60 min. Quantitative realtime polymerase chain reaction showed cul-5 mRNA levels were significantly increased in the brainstem and cerebellum, and decreased in the hypothalamus of rats as a result of hemorrhagic shock (n = 6) compared to sham-treated rats (n = 6). Cul-5 mRNA levels in the cerebral cortex, small intestine, kidney, liver, lung, or pituitary gland did not significantly change after hemorrhagic shock. This is the first report of cul-5 mRNA regulation by hemorrhagic shock. Evidence indicates this protein may have a regulatory role in ubiquitin-proteasomal protein degradation in response to hemorrhagic shock. PMID- 12635526 TI - Unruptured intracranial aneurysms in patients with transient cerebral ischemic episodes. Optional managements and literature review. AB - Clinical experience about the management of patients with an unruptured intracranial aneurysm (UIA) presenting transient cerebral ischemic attacks (TIAs) is usually restricted to a few case reports. Some authors assume tha it is safe to perform an endarterectomy without acutely increasing the risk of aneurysm rupture while others reported a post-operative aneurysmal bleeding even for aneurysms smaller than 10 mm between some days and several months after endarterectomy. The aim of this paper is to analyse the factors involved in the particular ischemic episodes, the examination methods to be employed, and the optional management for each individual case on a literature review and own experience-based data. PMID- 12635527 TI - The efficacy of non-contrast helical computed tomography in the prediction of urinary stone composition in vivo. AB - The composition of urinary stones was predicted on the basis of the differences in their Hounsfield unit (HU) densities as detected by in vivo non-contrast helical computed tomography (NCHCT). A total of 87 stones, which had been obtained from patients admitted to the emergency department with acute flank pain, were analysed biochemically. These stones had been visualized by NCHCT before spontaneous passage, ureteroscopic extraction or surgical removal, and their HU densities had been recorded. The HU densities of the stones were compared with the results of biochemical analysis. The mean HU densities of the stones composed of calcium oxalate (n = 54), struvite (n = 19) and uric acid (n = 14) were 812 +/- 135, 614 +/- 121 and 413 +/- 143, respectively. The differences between the mean HU densities of these three stone types were statistically significant. This initial clinical study demonstrated that in vivo NCHCT analysis may be used to predict the composition of urinary stones. PMID- 12635528 TI - Apoptosis in normal tissues induced by anti-cancer drugs. AB - To investigate the damage mediated by anti-cancer drugs in normal cells, we examined the effect of such drugs on apoptosis of normal cells of the small intestinal epithelium and the bone marrow by in situ DNA end-labelling and transmission electron microscopy. Mice received a single dose of 5-fluorouracil (5-FU) or cisplatin, or repeated daily doses of 5-FU for 7 days. In mice treated with a single dose of 5-FU 50 mg/kg or cisplatin 5 mg/kg, the number of apoptotic cells appearing in the small intestine 12 h after injection was relatively small, but increased steadily reaching a peak after 36 h and then decreasing to close to that in the control group by 48 h. In bone marrow cells, results were similar in mice treated with single doses of 5-FU 50 mg/kg but apoptosis increased much less in those treated with cisplatin 5 mg/kg. The proportion of apoptotic cells reached peak values earlier at higher concentrations of 5-FU or cisplatin both in small intestine and in bone marrow. In the mice treated repeatedly with 5-FU 50 mg/kg, the proportion of apoptotic small intestinal epithelial cells reached a succession of peaks at 48-h intervals. Mice treated repeatedly with 5-FU 50 mg/kg also showed a rapid increase in diarrhoea symptoms and a steady decrease in the height of villi. Our results suggest it may be possible to prevent the side effects of anti-cancer drugs by inhibiting apoptosis in the gastrointestinal tract. PMID- 12635529 TI - The importance of pulmonary artery circulation during cardiopulmonary bypass. AB - This study sought to determine changes in transpulmonary difference in blood cells and alveolar-arterial oxygen (A-aO2) gradient when pulmonary artery circulation was obstructed in patients undergoing coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB). Eighteen patients were divided into group A (control group; X-clamp placed on aorta, n = 9) and group B (pulmonary ischaemia group; X-clamp placed on aorta and pulmonary artery, n = 9). Haematological parameters were compared before CPB and up to 90 min after declamping. A-aO2 gradient differences were compared before and 2 h and 6 h after declamping. A transpulmonary increase in leucocyte levels normalized after 60 min in group A but remained higher in group B. A transpulmonary increase in neutrophils normalized after 60 min in group A and 90 min in group B. Increased lymphocyte levels normalized after 30 min in group A and 90 min in group B. A-aO2 gradient was determined as: group A (294.8 +/- 74.3) and group B (321.2 +/- 73.3) before X-clamping; group A (132.7 +/- 22.7) and group B (236.6 +/- 41.5) 2 h after declamping; and group A (72.2 +/- 22.7) and group B (189.4 +/- 88.9) 6 h after declamping. When pulmonary artery circulation was obstructed during the X clamping period, leucocyte, neutrophil and lymphocyte sequestration within both lungs increased, and an increased A-aO2 gradient was observed because of tissue damage. To prevent post-operative complications, precautions to maintain normal pulmonary artery circulation are recommended. PMID- 12635530 TI - Histological assessment of the effect of alpha-tocopherol on fracture healing in rabbits. AB - To investigate the effect of alpha-tocopherol (vitamin E) on fracture healing in rabbits, two groups of 10 rabbits were either injected with alpha-tocopherol (treated) or untreated (controls). The right femurs of both groups were fractured, and the treated group were injected intramuscularly with 20 mg/kg alpha-tocopherol daily for 5 days starting on the day of fracture. After 21 days, histological sections of the fractured region were examined and scored. Fracture healing had progressed further in the alpha-tocopherol group than in the control group. A statistically significant difference between the histological grading of fracture healing in the two groups was found. This difference may result from an antioxidant (alpha-tocopherol) effect on free oxygen radicals in the fracture area. We conclude that alpha-tocopherol may affect fracture healing favourably and might be useful as a therapeutic agent in clinical fracture management. PMID- 12635531 TI - Sedation for outpatient endometrial biopsy: comparison of remifentanil-propofol and alfentanil-propofol. AB - The purpose of the study was to compare the clinical activity of remifentanil or alfentanil under propofol sedation with regard to respiratory rate, sedation and recovery rate when used for outpatient endometrial biopsy. Patients were randomized to receive intravenously either bolus remifentanil 0.4 microgram/kg and propofol 1 mg/kg in the remifentanil group (n = 30), or bolus alfentanil 20 micrograms/kg and propofol 1 mg/kg in the alfentanil group (n = 30). Patients were monitored for heart rate, systolic and diastolic arterial pressure, peripheral O2 saturation (SpO2), respiration rate, and Aldrete sedation score. Pulse oximetry was used to monitor heart rate and SpO2 during endometrial biopsy. Apnoea was observed in five patients from the remifentanil group, and in three patients from the alfentanil group. The groups did not differ with regard to apnoea incidences. Times were recorded for orientation and Aldrete score > 8, and were similar between the two groups (13.20 +/- 3.64 min and 14.0 +/- 3.87 min in the remifentanil group, 14.7 +/- 3.64 min and 15.9 +/- 3.15 min in the alfentanil group, respectively). The sedative and analgesic combination of remifentanil propofol does not offer any advantages compared with a combination of alfentanil propofol with regards to respiration and recovery during sedation for outpatient endometrial biopsy. PMID- 12635533 TI - The beneficial effect of melatonin on chronic cyclosporin A nephrotoxicity in rats. AB - Cyclosporin A (CsA)-induced nephrotoxicity may be the consequence of oxidative stress. Anti-oxidant agents could be useful in reducing CsA toxicity. In this light microscopy study, tubular dilatation, atrophy, vacuolization and tubulointerstitial fibrosis were observed in rats given CsA, whereas in rats given CsA plus melatonin, no histological changes occurred. It is concluded that melatonin could be useful for reducing the nephrotoxic effects of CsA. PMID- 12635532 TI - Comparison of immunological and endocrinological markers associated with major depression. AB - Natural-killer-(NK)-cell activity and blood levels of interleukin 2 (IL-2), dehydro-epiandrosterone (DHEA), DHEA sulphate (DHEA-S) and cortisol were measured in 17 patients with major depression and 10 control subjects. Depression severity was evaluated using the Zung Self-rating Depression Scale. NK-cell activity and IL-2 levels were measured using a chromium-51 release test and an enzyme-linked immunosorbent assay, respectively. Radio-immunoassays were used to measure serum cortisol, DHEA and DHEA-S. As would be expected, patients with major depression had a higher score on the Zung Self-rating Depression Scale than healthy controls. Compared with controls, NK-cell activity and levels of cortisol and DHEA were reduced in patients with major depression, whereas IL-2 levels were increased. No difference was observed in DHEA-S levels between patients and controls. A reduction in NK-cell activity and DHEA levels, and an increase in IL 2 levels appear to be associated with major depression. Whether these changes are the cause or the consequence of the depression remains to be determined. PMID- 12635535 TI - Reiter's syndrome associated with HLA-B51: a case report. AB - A 32-year-old Japanese man developed polyarthritis with mild fever and conjunctivitis. Clinical assessment indicated non-specific arthritis, aseptic pyuria induced by infection with Chlamydia, and conjunctivitis. He was diagnosed with reactive arthritis (Reiter's syndrome). Serotyping of human leucocyte antigen (HLA) class I and II revealed positivity for B51(5), A2, A33(19), B44(12), Cw1, DR4 and DR6, but B27 was negative. He was treated with a combination of doxycycline, oral prednisolone, diclofenac sodium and salazosulphapyridine. Fever and arthralgia improved and he became negative for anti-Chlamydia immunoglobulin (Ig) A and IgG antibodies. HLA-B51 may be involved in the pathogenesis of Reiter's syndrome in this Japanese patient. PMID- 12635534 TI - Efficacy and safety of azithromycin as monotherapy or combined with metronidazole compared with two standard multidrug regimens for the treatment of acute pelvic inflammatory disease. AB - The objective of the study was to compare the efficacy of azithromycin, alone or with metronidazole, versus two standard multidrug regimens for the treatment of acute pelvic inflammatory disease (PID). Patients with PID were treated with once daily intravenous (i.v.) azithromycin 500 mg for 1 day or 2 days followed by once daily azithromycin 250 mg orally for a total of 7 days, alone or with three-times daily metronidazole 400 mg or 500 mg i.v. then orally for a total of 12-14 days. The comparators were either metronidazole + doxycycline + cefoxitin + probenecid or doxycycline + amoxycillin/clavulanate given at standard recommended doses for up to 21 days. In total, 309 patients were treated for PID. The diagnosis was confirmed laparoscopically in 74.8% of patients. Rates of clinical success for azithromycin, alone (97.1%) or with metronidazole (98.1%), were comparable to those for the comparator regimens (94.6%). Eradication rates for Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma hominis and anaerobes were also comparable for each of the treatment groups. Both azithromycin regimens were well tolerated. In conclusion, azithromycin, alone or with metronidazole, provides a shorter, simpler treatment option for the successful management of acute PID. PMID- 12635536 TI - Shifting perspectives on gambling and addiction. PMID- 12635537 TI - The neurobiology of pathological gambling: translating research findings into clinical advances. PMID- 12635538 TI - Genetics of pathological gambling. AB - Pathological gambling (PG) is an impulse control disorder and a model 'behavioral' addiction. Familial factors have been observed in clinical studies of pathological gamblers, and twin studies have demonstrated a genetic influence contributing to the development of PG. Serotonergic, noradrenergic, and dopaminergic dysfunction have been reported as biological factors contributing to the pathophysiology of PG. Molecular genetic techniques have been used to investigate the role of genetic factors in PG. Molecular genetic research has identified specific allele variants of candidate genes corresponding to these neurotransmitter systems to be associated with PG. Associations have been reported between pathological gamblers and allele variants of polymorphisms at dopamine receptor genes, the serotonin transporter gene, and the monoamine oxidase A gene. Although preliminary data suggest that some of these differences are gender-specific, more research needs to be performed to substantiate gender specific genetic contributions to the development of pathological gambling. The review of the current findings on genetics of PG suggests that liability to PG is in part mediated by genetic factors. Additional studies are needed to replicate and extend these findings, as well as to better understand the influence of specific allelic variants to differences in biological and behavioral functioning. PMID- 12635539 TI - Risky business: emotion, decision-making, and addiction. AB - Although metabolic abnormalities in the orbitofrontal cortex have been observed in substance dependent individuals (SDI) for several years, very little attention was paid to the role of this brain region in addiction. However, patients with damage to the ventromedial (VM) sector of the prefrontal cortex and SDI show similar behaviors. (1) They often deny, or they are not aware, that they have a problem. (2) When faced with a choice to pursue a course of action that brings an immediate reward at the risk of incurring future negative consequences, they choose the immediate reward and ignore the future consequences. Studies of patients with bilateral lesions of the VM prefrontal cortex support the view that the process of decision-making depends in many important ways on neural substrates that regulate homeostasis, emotion, and feeling. Parallel lines of study have revealed that VM cortex dysfunction is also evident in subgroups of individuals who are addicted to substances. Thus, understanding the neural mechanisms of decision-making has direct implications for understanding disorders of addiction and pathological gambling, and the switch from a controlled to uncontrolled and compulsive behavior. On the clinical front, the approach to treat addictive disorders has been dominated by a diagnostic system that focuses on behaviors, physical symptoms, or choice of drugs. The article emphasizes the concept of using neurocognitive criteria for subtyping addictive disorders. This is a significant paradigm shift with significant implications for guiding diagnosis and treatment. Using neurocognitive criteria could lead to more accurate subtyping of addictive disorders, and perhaps serve as a guide for more specific, and potentially more successful, behavioral and pharmacological interventions. PMID- 12635542 TI - [Pain in the legs and progressive neurological deterioration: tethered cord syndrome in adults]. AB - In four patients, women aged 58, 63 and 42 years, and a man aged 25 years, tethered cord syndrome was diagnosed. These patients suffered from progressive neurological symptoms, notably radiating pain in the legs in the first three patients, and muscle weakness and atrophy in the lower legs in the fourth patient. All four were treated surgically. Nowadays, the phrase tethered (spinal) cord is used for both occult and open closure defects of the neural tube. New neurological symptoms and signs can develop not only in childhood, but also in adults. Neurosurgical release of the tethered cord prevents further deterioration and often leads to significant subjective improvement of symptoms. PMID- 12635543 TI - [European ordinance on orphan drugs: changes and threats]. AB - The Orphan Drugs Act has been officially implemented in all countries of the European Union since the year 2000. The Act aims to promote the development of treatments for rare diseases (prevalence < 5:10,000). Successful therapies for such diseases are granted market-monopoly for 10 years, during which no me-too products are allowed. There are problems concerning approval and reimbursement. Drugs already in use can also be registered as orphan drugs for a specific indication. In addition to this, European approval for the market authorization of new drugs does not automatically mean that the costs of the drug are reimbursed within the Netherlands, whereas that might well be the case in other EU countries. However, a faster procedure with respect to the reimbursement of drug costs in the Netherlands may not lead to the responsibilities for the carrying out of additional trials being transferred to those handling the treatment. PMID- 12635544 TI - [The treatment of gallstone disease in the elderly]. AB - Gallstone diseases (asymptomatic, symptomatic and complicated) are frequently seen in the elderly; the prevalence increases proportionally with age. At higher ages (> 60 years) the presentation of symptomatic or complicated gallstone disease is frequently atypical. Complicated gallstone disease (especially cholecystitis and cholangitis) in the elderly is associated with high morbidity and mortality rates. The introduction of laparoscopic cholecystectomy has decreased the morbidity and mortality rates of symptomatic and complicated gallstone disease in the elderly; for elective procedures in particular, the risks hardly differ from those for younger patients. Percutaneous cholecystostomy is an effective and safe alternative for (laparoscopic) cholecystectomy in high risk patients with an acute cholecystitis. Endoscopic retrograde cholangiopancreaticography (ERCP) with sphincterotomy is also the treatment of choice for common bile duct stones in the elderly. After removal of common bile duct stones (whether or not accompanied by cholangitis or pancreatitis) a laparoscopic cholecystectomy should be performed, unless contraindications are present. PMID- 12635540 TI - Neurodevelopment, impulsivity, and adolescent gambling. AB - The prevalence of problem and pathological gambling in adolescence and young adulthood has been found to be two- to fourfold higher than in adulthood. Given that these high rates might predict future increases across all age groups, it is important to explore the causes of the elevated rates of problem and pathological gambling among youths. This article reviews evidence for a neurobiological basis for adolescent vulnerability to problem and pathological gambling behaviors. We propose that a common trait motif of impulsivity might underlie phenomenology of pathological gambling, commonly comorbid psychiatric disorders, and related aspects of adolescent behavior. Recent advances in understanding the brain mechanisms involved in motivation, reward, and decision-making allow a discussion of neural circuitry underlying impulsivity. Emerging data indicate that important neurodevelopmental events during adolescence occur in brain regions associated with motivation and impulsive behavior. We hypothesize that immaturity of frontal cortical and subcortical monoaminergic systems during normal neurodevelopment underlies adolescent impulsivity as a transitional trait-behavior. While these neurodevelopmental processes may confer advantage by promoting a learning drive for optimal adaptation to adult roles, they may also confer an increased vulnerability to addictive behaviors such as problem and pathological gambling. An exploration of the developmental changes in neural circuitry involved in impulse control has significant implications for understanding adolescent behaviors and treating problem and pathological gambling among youths. PMID- 12635545 TI - [Sarcoidosis: immunopathogenesis and the potential of immunotherapy]. AB - Sarcoidosis is a multi-systemic disease of unknown aetiology, immunopathologically and histologically characterised by a macrophage/T-helper I cell-mediated non-caseating granulomatous inflammation process. In the development of granulomas, an imbalance between pro-inflammatory and anti inflammatory cytokines plays an important role, possibly initiated by an as yet unidentified (exogenous) stimulus. The clinical outcome of the disease process is unpredictable, and appears to be determined by the cytokine production of the inflammatory cells in the granuloma. Fifty to seventy percent of the patients recover without medication within a period of time which cannot be predicted, the other 30 to 50% of patients are treated during the course of the disease with corticosteroids (with varying degrees of success), either alone or in combination with cytostatic or immunosuppressive agents. Based on recent developments in the field of immunomodulation and on current knowledge of the immunopathogenesis of sarcoidosis, there appear to be opportunities for specific immunotherapy which should be evaluated in controlled studies. PMID- 12635541 TI - Advances in the pharmacological treatment of pathological gambling. AB - In the present paper we discuss the current status of drug treatment for pathological gambling and the scientific rationales underlying the various pharmacological approaches. Specifically, we summarize the treatment study results of serotonin reuptake inhibitors, mood stabilizers, opioid antagonists, and atypical antipsychotics in pathological gambling. We also discuss dosage strategies, the duration of treatment, issues surrounding medication compliance, and approaches to treatment-refractory pathological gambling, such as pharmacological and behavioral augmentation. PMID- 12635546 TI - [Diagnostic image (123). A man on oxygen therapy and facial burns. First and second degree burns in the face after a flame during nasal oxygen administration]. AB - A 66-year-old man was hospitalised because of pneumonia and chronic obstructive pulmonary disease. He suffered from facial burns when he tested a cigarette lighter whilst on oxygen therapy by nasal prongs. PMID- 12635547 TI - [From gene to disease; apolipoprotein E2 and familial dysbetalipoproteinemia]. AB - Familial dysbetalipoproteinaemia is an autosomal recessive, hereditary disorder of lipid metabolism caused by mutations in the apolipoprotein E gene. Homozygosity for apoE2 (1 in 170 persons) causes type III hyperlipoproteinaemia in less than 20% of the adult E2 homozygotes. The patients may present with typical skin lesions and have elevated plasma levels of cholesterol and triglycerides, mainly in very-low-density lipoprotein remnants and intermediate density lipoproteins. The disorder is associated with peripheral and coronary artery disease. Additional genetic and environmental factors are necessary for the expression of this hyperlipoproteinaemia. Hyperinsulinaemia and defects in genes involved in the hydrolysis of triglycerides are associated with this disorder. PMID- 12635548 TI - [Evaluation of the Rotterdam guideline for referral in case of chronic hepatitis B: improvement possible via a shortening of the referral chain and better distribution of information]. AB - OBJECTIVE: Evaluation of the Rotterdam guideline for referring patients with chronic hepatitis B virus (HBV) infections from primary to specialist care for diagnosis and treatment. DESIGN: Retrospective. METHOD: Whether or not the guideline was followed correctly was determined in patients with chronic hepatitis B who were reported to the Municipal Health Service (GGD) in Rotterdam in 1998 and 1999. This was done by a study of their files and by questioning their general practitioners by phone. RESULTS: During the study period, 376 cases of chronic hepatitis B were seen at the GGD; 32% of the patients dropped out during the referral trajectory. Drop-out took place at three different times: 13% during the process of deciding whether the patient should be referred according to the guideline, 12% during the consultation period at the GGD, and 7% after consultation at the GGD and before first contact with a specialist (via referral from the general practitioner). The reasons for dropping out were either procedural factors, such as missing information in the patient file and unclear management by the GGD, and personal factors such as failure of the general practitioner or the patient to comply with the recommendations. CONCLUSION: The guideline for the referral of chronic hepatitis B patients functioned well but implementation can be improved, since some patients did not reach the specialist. Improvement would be made possible by shortening the referral chain and by giving more information to patients and general practitioners about hepatitis B and its potential consequences. PMID- 12635549 TI - [Percutaneous insertion of a left ventricular assist device for cardiogenic shock in a patient with a myocardial infarction]. AB - A 54-year-old man who was admitted due to cardiogenic shock as a result of a large anterior myocardial infarction 3 days previously was about to die, despite reperfusion therapy, application of an intra-aortic balloon pump, mechanical ventilation and maximal medical therapy. After insertion of a percutaneous left ventricular assist device, the patient was haemodynamically stable. After 11 days, the assist device was weaned and was removed. One day later, the patient died due to progressive heart failure. This case shows that a percutaneously inserted left ventricular assist device is effective in patients with severe, refractory cardiogenic shock, and is relatively simple to insert in the heart catheterisation room. However, it is still not clear what the recovery possibilities of the heart are following a large myocardial infraction, which factors may influence this recovery, and what the applicability of such a ventricular assist device might be in bridging the recovery period. PMID- 12635550 TI - [Primary meningococcal-monoarthritis (Neisseria meningitidis serotype C) in a child]. AB - A 26-month-old girl had a painful, swollen right knee, accompanied by fever and vomiting. She had had an upper respiratory tract infection for a number of days. Following a positive culture of synovial fluid, monoarthritis caused by Neisseria meningitidis serotype C without other signs of illness was diagnosed. She recovered completely after one joint aspiration and intravenous and oral antibiotic therapy. N. meningitidis serotype C infections without meningitis or septicaemia are rare, but should form part of the differential diagnosis of septic monoarthritis. PMID- 12635551 TI - [Similarities and differences between the euthanasia laws in Belgium and the Netherlands]. AB - Recently, a law on euthanasia has been adopted in both the Netherlands and Belgium. In both countries euthanasia has been legalized under strict conditions and after confirmation with a notification procedure. Although both laws are similar, the Belgian law is more extensive on the requirements of prudent practice. On the other hand, in Dutch society the norm-setting on euthanasia has been more widely developed through jurisprudence. Nevertheless, we expect that the implementation of the new law and the notification procedure in Belgium will be more difficult than in the Netherlands. In order to promote, safeguard and guarantee the quality of the euthanasia practice, the present euthanasia notification procedure in the Netherlands is supplemented with feedback to the physicians. The strict anonymous procedure in the Belgian notification procedure prevents this possibility. Therefore, Belgian physicians will not be supported by the notification procedure to improve their knowledge and skills in euthanasia. PMID- 12635552 TI - [Information provision for the biomedical sciences in the Netherlands faces an uncertain future]. PMID- 12635553 TI - [Information provision for the biomedical sciences in the Netherlands faces an uncertain future]. PMID- 12635555 TI - [The standard "Vaginal bleeding" of the Dutch College of General Practitioners]. PMID- 12635554 TI - [The added value of a structured evaluation of patients with vascular disease]. PMID- 12635556 TI - 2002--defining the 21st century II. PMID- 12635557 TI - Twinning research and practice guidelines in the management of addictions. PMID- 12635558 TI - Substance use disorders: sex differences and psychiatric comorbidities. AB - OBJECTIVE: This article reviews sex differences in psychiatric comorbidity among individuals with substance use disorders and, in particular, the clinical significance of these differences for treatment outcome among women. METHOD: We undertook a computerized search of major health care databases. To enhance the search, we drew prior relevant articles from the reference list. RESULTS: Women with alcohol and other drug use disorders present higher rates of psychiatric comorbidity, particularly mood and anxiety disorders, than do men. Moreover, the comorbid diagnosis, particularly of depression, is more often primary in women, while in men the comorbidity is more often secondary to the substance abuse diagnosis. In addition, there is evidence that psychiatric comorbidity is associated with distinct, sex-specific outcomes for substance use treatment. CONCLUSIONS: Sex differences in the clinical presentation of substance-dependent individuals with psychiatric comorbidity present specific treatment challenges and opportunities. PMID- 12635559 TI - Clinical aspects of substance abuse in persons with schizophrenia. AB - OBJECTIVE: To review the current knowledge on the problem of psychoactive substance abuse by persons with schizophrenia, with particular attention to issues of direct relevance to clinical practice. METHOD: The author examined the literature from the last 2 decades and data from studies in which he was involved. RESULTS: Schizophrenia sufferers show an elevated liability for substance abuse. Such comorbidity may derive from self-medication attempts, a common neuropathology for addiction and schizophrenia, the psychotogenic properties of certain drugs, or the influence of environmental factors. Among schizophrenia patients receiving treatment, substance misuse is associated with more severe symptoms and poorer therapeutic response. The presence of a chronic psychosis impedes treatment of the substance problem in traditional, nonpsychiatric addiction programs. Better outcomes are observed in integrated therapy services, where patients receive appropriate care for both conditions. CONCLUSION: Dual-pathology patients need comprehensive care with appropriate pharmacotherapy and psychosocial interventions. This treatment can be best provided within the context of a continuing care psychiatric service. PMID- 12635560 TI - Are there cognitive and behavioural approaches specific to the treatment of pathological gambling? AB - OBJECTIVE: Treatment approaches for pathological gambling have been modelled after preexisting substance addiction models. This paper reviews cognitive behavioural models in a search for original insights that are specific to gambling treatment. METHOD: A computerized search of major health care databases (Medline and PsycINFO) was performed. RESULTS: New cognitive-behavioural approaches to the treatment of pathological gambling provide 3 original additions to the traditional multimodal treatment of addictions: cognitive restructuring, in vivo exposure, and imaginal desensitization. Other cognitive-behavioural techniques, such as relapse prevention, problem solving, and social skill training, are shared by gambling treatment and addictions treatment. CONCLUSIONS: When treating pathological gamblers, clinicians must consider introducing techniques to address cognitive distortions related to gambling. Also, cue exposure--whether in vivo or imaginal--may help deal with urges prompted by such cues. The blending of these new techniques into a multimodal addiction treatment potentially balances the rational and external orientation of the cognitive behavioural approach with interpersonal and introspective components of the traditional addiction treatment. PMID- 12635561 TI - The relation between memory of the traumatic event and PTSD: evidence from studies of traumatic brain injury. AB - OBJECTIVE: This paper focuses on the relation between memory and posttraumatic stress disorder (PTSD). More specifically, it addresses the debate regarding the role of memory of the traumatic event in the development of PTSD. Traumatic brain injury (TBI) is used as a naturally occurring model for traumatic exposure that is often associated with memory impairment. METHOD: We present a critical review of the literature on studies assessing the relation between TBI and PTSD, with a focus on memory of the traumatic event as a critical factor. We also discuss results from recent studies conducted by our group. RESULTS: The literature review offers an inconclusive picture wherein a significant proportion of the studies indicate that PTSD and TBI are mutually exclusive, especially in individuals who exhibit lack of memory for the traumatic event. This finding supports the possibility that lack of memory may protect against the development of PTSD. However, some studies show that PTSD does occur in patients with head injury, suggesting that PTSD may develop in TBI survivors--even in those who cannot remember the traumatic event. Generally speaking, though, the overall balance of the findings (including our own findings) seems to support the possibility that, in subjects with TBI, impaired memory of the traumatic event is associated with reduced prevalence of PTSD. CONCLUSIONS: The suggestion that amnesia regarding the traumatic event may protect against the development of PTSD has both theoretical and practical importance. This review focused on the case of traumatic brain injury as a model for impaired memory for the traumatic event. However, it still remains to be proven that the conclusions based on these findings are generalizable beyond the case of TBI. While some patients with posttraumatic amnesia do develop PTSD despite lack of memory for the traumatic event, the majority of those who lack memory for the event seem to be protected from developing the disorder. Nevertheless, based on this assumption, we suggest that pharmacologic disruption of newly acquired--or even old--traumatic memories, which has been shown to be possible in animals, might therapeutically benefit trauma survivors. PMID- 12635562 TI - Evolutionary perspectives on schizophrenia. AB - The theory of evolution may be relevant to psychiatric disorders. Evolution reflects changes in genes throughout time. Thus, evolutionary forces can shape any phenotype that is genetically rooted and that possesses a long history. Schizophrenia is likely an ancient condition with a substantial genetic component. Since the 1960s, several researchers have applied evolutionary principles to the study of schizophrenia. In general, schizophrenia is either viewed as an evolutionary advantageous condition or as a disadvantageous byproduct of normal brain evolution. This paper reviews major evolutionary explanations--historical and current--that speculate on the possible origins of schizophrenia. PMID- 12635563 TI - Effect of a new casino on problem gambling in treatment-seeking substance abusers. AB - Problem gambling rates are frequently found to be higher in those who abuse substances than in the general population, and this group represents a well established high-risk population for developing the disorder. In this study of 853 residential substance abusers, approximately 10% scored in the problem gambling range on the South Oaks Gambling Screen (SOGS). On most descriptive variables, these subjects appeared to be similar to substance abusers who do not have gambling problems. However, they tended to participate in more gambling behaviours and had more relationships with individuals who also gambled. There is some evidence that the introduction of a new casino in the community increased the SOGS scores for subjects who gambled most frequently on such casino-related gaming as slot machines, cards, and casino games. PMID- 12635564 TI - The thought disorder questionnaire. AB - OBJECTIVE: To describe the development of the Thought Disorder Questionnaire (TDQ), including data on reliability and validity, and to explain the questionnaire's feasibility and diagnostic accuracy. METHODS: The TDQ has 6 scales, each with 10 items (on a scale from 0 [for never] to 4 [for always]). The 6 scales measure content of thought, control of thought, orientation, perception, fantasy, and symptoms. RESULTS: The TDQ is a reliable 60-item, self-report questionnaire that measures the quantity and quality of disordered thinking in patients with mental disorders. It has established reliability and validity. CONCLUSION: The TDQ's clinical and research utility remains to be determined. PMID- 12635565 TI - The index manic episode in juvenile-onset bipolar disorder: the pattern of recovery. AB - OBJECTIVE: Recent studies of patients with juvenile bipolar disorder report low rates of recovery and high rates of chronicity. However, we lack data on the short-term outcome. This study examines the pattern of recovery from the index episode in an aggressively treated juvenile sample. METHOD: We assessed 25 subjects (< 16 years) with a diagnosis of mania, using the Diagnostic Interview for Children and Adolescents-Revised) (DICA-R), Young Mania Rating Scale (YMRS), and Children's Global Assessment Scale (CGAS) at intake and at 3 and 6 months. We studied the time taken to recover from the index episode, the level of functioning, and the factors predicting them. RESULTS: After 6 months, 24 (96%) subjects had recovered from the index manic episode. The median time to recovery was 27 days. Total episode length was significantly longer among those with previous affective episodes. CONCLUSIONS: The findings suggest that juvenile onset mania has high rates of recovery and low rates of chronicity. These differences from the existing literature need further exploration. PMID- 12635566 TI - Validation of a French version of the impact of event scale-revised. AB - OBJECTIVE: This report presents a French translation and validation of the Impact of Event Scale-Revised (IES-R) in a population of women exposed to a natural disaster during or preceding pregnancy. METHOD: A total of 223 francophone women who were either pregnant at the time of the 1998 ice storm or who became pregnant shortly thereafter completed the IES-R and other questionnaires 6 months after the disaster. RESULTS: The French IES-R has good internal consistency, with alpha coefficients ranging from 0.81 to 0.93 for its 3 subscales and total score. The test-retest reliability of the scale, although examined with another sample, proved to be satisfactory, with correlation coefficients ranging from 0.71 to 0.76 for its 3 subscales and total score. Its convergent validity with perceived life threat and general psychiatric symptoms was judged to be marginally acceptable. Finally, a principal components analysis was conducted and a 3-factor solution, which explained 56% of the variance, was retained: a hyperarousal factor (7 items), an avoidance factor (6 items), and an intrusion factor (6 items). CONCLUSIONS: The French version of the IES-R has satisfactory internal validity and test-retest reliability. Further, the factor structure of the translation was similar to the proposed theoretical structure of the IES-R. PMID- 12635567 TI - Re: Atypical antipsychotics mechanisms of action. PMID- 12635568 TI - Re: "Cades disease" and beyond. PMID- 12635569 TI - Quetiapine-induced leucopenia: possible dosage-related phenomenon. PMID- 12635570 TI - Atypical neuroleptic malignant syndrome with clozapine and subsequent haloperidol treatment. PMID- 12635571 TI - Stachybotrys chartarum (chartarum = atra = alternans) and other problems caused by allergenic fungi. AB - Stachybotrys chartarum is a cellulose-decaying fungus with worldwide distribution. It grows well at room temperature and with humidity above 93%. S. chatarum requires special media high in cellulose and low in sugar and nitrogen to compete with Penicillium and Aspergillus. Ninety percent of field-collected spores are not culturable. S. chartarum can produce macrocyclic trichothecenes but is highly dependent on strain and environmental conditions. In strains implicated in mycotoxicosis, not all produce detectable trichothecenes. Therefore, the presence of S. chartarum is not proof of toxin presence. Trichothecenes are potent inhibitors of protein and DNA synthesis. By the inhalation route, occupational stachybotrytoxicosis causes chest and upper airway symptoms, fever, leucopenia, dermatitis; starts in 2-3 days of exposure; and lasts 3 weeks. Investigation of the environment of the cluster of pulmonary hemorrhage in 10 infants in Cleveland, Ohio, and similar cases elsewhere are presented. The Centers for Disease Control and Prevention considers S. chartarum a serious health threat. However, even though there are now techniques of measuring S. chartarum conidia and estimating trichothecene mycotoxin in indoor air samples, no standards exist that relate to health effects. Those standards available are numerical or comparison of indoor/outdoor counts or both. Upper limit of noncontaminated indoor environment is 100-1000 colony-forming units (CFU) m3. There is no compelling evidence that exposures expected in most mold contaminated indoor environments are likely to result in measurable health effects. However, when the health care worker suspects a problem in the home environment, a questionnaire and home visit may be helpful. High indoor exposures are associated with infrequent ventilation or vacuuming, pets, visible mold, and old carpets. To screen the indoor air, an experienced pollen and mold counter could use a Burkard personal air sampler. Health-based exposure standards for molds and mycotoxins do not exist. When available data indicate extremely high mold levels, cleanup consisting of removal of all contaminated material, cleaning accessible heating, ventilation, and airconditioning parts and filters, and preventive maintenance are indicated. There is a brief summary of the diseases of plants, animals, and humans caused by several common allergenic fungi and the mycotoxins they produce. PMID- 12635572 TI - Primary acquired cold urticaria. AB - Primary acquired cold urticaria (ACU) is the most common type of cold urticaria characterized by rapid onset of pruritic hives, swelling, and possible severe systemic reactions including hypotension and shock after cold exposure. Primary ACU is diagnosed by history of such symptoms, a positive immediate cold-contact stimulation test, and negative laboratory evaluation for underlying systemic disorders. Clinicians should be aware that patients with ACU may be susceptible to life-threatening systemic reactions especially during aquatic activities and that proper patient education is extremely important. This article reviews the clinical presentation, pathogenesis, diagnosis, and management of primary ACU. PMID- 12635573 TI - Hypersensitivity to molds in New York City in adults who have asthma. AB - Molds have been linked epidemiologically to asthma as a key aeroallergen in several studies. Other allergens such as cockroach have been linked to asthma in New York City (NYC). To our knowledge, however, the pattern of mold hypersensitivity has never been examined systematically in the NYC area. Thus, we sought to determine the association between mold hypersensitivity and asthma in a large group of ambulatory patients evaluated for allergic disease for the years 1993 through 2001 at a single medical center. Serological testing for mold specific immunoglobulin E (IgE) as well as IgE specific for other aeroallergens was performed and the associations between allergen-specific IgE and the presence of asthma were examined using bivariate and multivariate analysis. Factor analysis showed that three distinct groupings of aeroallergen-specific IgE existed within the panel of allergens used. Group 1 consisted of cat dander and dust mites (Dermatophagoides farinae). Group 2 consisted of tree, grass, and ragweed pollen. Group 3 consisted of the Deuteromycetes molds, Alternaria tenuis, Aspergillus fumigatus, and Cladosporium herbarum. Patients with asthma had a highly significant increase in the incidence of hypersensitivity to cat/dust mites and to the molds. Multivariate analysis showed that the presence of hypersensitivity to either A. tenuis or C. herbarum had a significant independent association with asthma after adjustment for cat/dust mite hypersensitivity and after adjustment for other clinical factors. On the other hand, pollen hypersensitivity was not associated independently with asthma. Mold hypersensitivity was strongly correlated with hypersensitivity to cat or dust mites in patients who did not have asthma but not in patients who did have asthma. In the NYC area, recent pollen and spore counts show that mold spores are measurable in at least 75% of the year. Thus it is conceivable that mold hypersensitivity plays a contributing and independent role in initiating or perpetuating the allergic response in patients with asthma in the New York area. PMID- 12635574 TI - Production of TNF-alpha by peripheral blood mononuclear cells through activation of nuclear factor kappa B by specific allergen stimulation in patients with atopic asthma. AB - Stimulation by specific allergens induces inflammatory cytokine production from peripheral blood mononuclear cells (PBMCs) in patients with atopic asthma, but the mechanism remains unknown. PBMCs were collected from six patients with atopic asthma with an immunoglobulin E-radioallergosorbent test score to Dermatophagoides farinae of > or = 4 and six nonatopic healthy subjects (score = 0) using a dish adhesion method after density gradient centrifugation. CD23 expression in PBMCs was analyzed by the fluorescence-activated cell sorting method. PBMCs were incubated with D. farinae or lipopolysaccharide, and production of tumor necrosis factor (TNF) alpha into the supernatant was measured by enzyme-linked immunosorbent assay. After incubation, immunostaining nostaining of the PBMCs with anti-nuclear factor kappa B (NF-kappa B) antibody (anti-p65 antibody against p65 as the subunit of NF-kappa B) was performed, and NF-kappa B activation in extracted nuclear protein was examined by electrophoretic mobility shift assay. CD23 expression was significantly higher in PBMCs from patients with asthma than in the controls (p < 0.01). There was no significant difference in TNF-alpha production by lipopolysaccharide stimulation between the two groups, but D. farinae-specific TNF-alpha production was significantly higher in subjects with asthma than in the controls (p < 0.05). A significant translocation of NF kappa B to nuclei by D. farinae stimulation was observed in cells from subjects with asthma (p < 0.01). Our results indicated that TNF-alpha production was induced by D. farinae in PBMCs of patients with atopic asthma by the activation of NF-kappa B via CD23. In patients with atopic asthma, CD23-mediated signals may cause proinflammatory cytokine production, which may lead to airway inflammation. PMID- 12635575 TI - Interleukin-2 induces peroxide production by primed normodense eosinophils of patients with asthma. AB - In this study we assessed, by flow cytometry, the effect of interleukin 2 (IL-2) on the oxidative burst of normodense eosinophils (Eos's) isolated from 15 patients with moderately severe extrinsic asthma and 17 controls. We found that IL-2 significantly induced peroxide (H2O2) production in normodense Eos's from patients with asthma on a time kinetics study. This rise was higher in patients with immunoglobulin E levels > 180 IU/mL versus normal immunoglobulin E values. The effect of IL-2 was partially blocked by using anti-Tac antibody. In contrast, IL-2 decreased H2O2 production in normodense Eos's from controls. Cell surface expression of CD25, CD122, CD132, and CD69 were also determined and no statistical differences were found between both groups. In conclusion, IL-2 is able to increase H2O2 production by normodense Eos's isolated from patients with asthma and it may contribute to bronchial epithelium damage and chronic inflammation. PMID- 12635576 TI - Expression of matrix proteins in an in vitro model of airway remodeling in asthma. AB - Acute asthma is characterized by a decrease in the pH of the exhaled breath condensate and bronchoconstriction. These perturbations may injure the epithelium in a chronic, intermittent pattern, leading to subepithelial fibrosis. We used an in vitro three-dimensional model of the bronchial mucosa to elucidate the response to a repeated chemical or physical insult to the epithelium in the postcontraction phase. We used enzyme-linked immunosorbent assay and reverse transcriptase--polymerase chain reaction to assess the production of the following proteins: matrix metalloproteinase (MMP) 3, MMP-9, tissue inhibitor of MMP-1, transforming growth factor beta 1, thrombospondin 1, tenascin, and fibronectin. The presence of the epithelium enhanced the degree of tissue contraction (50.1 +/- 4.4% of original area versus 75.4 +/- 2.3%). In the absence of injury, tenascin, fibronectin, MMP-3, and tissue inhibitor of MMP-1 are actively expressed. However, the chronic chemical wound markedly inhibited the expression of all proteins. We conclude that the epithelium, wound type, and age of the tissue (contracting versus postcontraction) impact the expression of key proteins in an in vitro model of subepithelial fibrosis in asthma. PMID- 12635577 TI - Treatment effectiveness of inhaled corticosteroids and leukotriene modifiers for patients with asthma: an analysis from managed care data. AB - We compared measures of treatment effectiveness when inhaled corticosteroids (ICSs) or leukotriene modifiers (LMs) were used as controller monotherapy for asthma. Asthma patients aged 6-55 years initiating ICS or LM monotherapy between July 1998 and June 1999 (index prescription) were identified using a managed care claims database. Asthma-related hospitalizations, emergency department (ED) visits, and use of short-acting beta-agonists and oral corticosteroids (OCSs) were assessed as proxies for treatment effectiveness. Propensity score was used to adjust for baseline differences between treatment cohorts. The change in the annual rate of claims from the preindex to postindex period for each measure was compared across treatment groups. Logistic regression models of the postindex composite events (hospitalization and/or ED) and OCS use were estimated. Nine hundred sixty patients were initiated on LMs (n = 153) and ICSs (n = 807). The mean annual rate of claims for OCSs increased in the ICS group (0.2) but was unchanged in the LM group (adjusted mean difference in change, 0.2; 95% CI, 0.05 0.4; p = 0.01). The mean annual rate of claims for short-acting beta-agonists increased in both the ICS and LM groups by 1.1 and 0.5, respectively (adjusted mean difference in change, 0.6; 95% CI, -0.06. 1.1; p = 0.08). Similar changes in annualized rates of claims for hospitalizations and ED visits were observed between treatment groups. In logistic regression models, greater odds of postindex OCS use was observed among the ICS group (odds ratio for ICS versus LM = 1.7; 95% CI, 1.04-2.8; p = 0.04). No association between treatment groups and postindex hospitalization and/or ED events was observed. In this managed care population, patients treated with ICSs or LMs had similar measures of treatment effectiveness, as measured by asthma-related health care resource use. PMID- 12635578 TI - The effect of topical nasal fluticasone on objective sleep testing and the symptoms of rhinitis, sleep, and daytime somnolence in perennial allergic rhinitis. AB - Recent data suggested that daytime somnolence in patients with allergic rhinitis was secondary to disrupted sleep caused by nasal congestion. Medications, which decreased congestion, would be expected to improve sleep and daytime somnolence. Previously, we showed that nasal steroids improved all three symptoms. Presently, we have not performed objective sleep testing to determine if there is a correlation between subjective improvement of congestion, sleep, and daytime somnolence. The objective of this 8-week, double-blind, placebo-controlled study was to determine if topical nasal fluticasone is effective at decreasing subjective congestion and daytime somnolence and improving sleep and if this improvement correlated with a change in overnight sleep testing (polysomnography). We recruited 32 subjects with perennial allergic rhinitis and randomized them in a double-blinded, cross-over fashion, to receive placebo or fluticasone (50 micrograms a spray), 2 sprays each side everyday, using Balaam's design. Questionnaires, quality of life instruments, daily diary, Epworth Sleepiness Scale, and an overnight sleep test with polysomnograms were used as tools. The last 2 weeks of each 4-week treatment period were summarized, scored, and compared by PROC MIXED in SAS. Correlations between arousals on sleep tests and subjective tests were performed. Fluticasone improved subjective sleep when compared with placebo (p = 0.04); however, there was no difference in the apnea/hypopnea index in those that were treated. Daytime sleepiness and fatigue were decreased by > 10% in the treated group; however, this was not statistically significant. However, fluticasone used at approved doses improves subjective sleep in patients with perennial allergic rhinitis without a change in the apnea/hypopnea index. PMID- 12635580 TI - Effect of staphylococcal enterotoxin B on specific antibody production in children with atopic dermatitis. AB - Exotoxins secreted by Staphylococcus aureus have been identified as a possible trigger factor in atopic dermatitis (AD). We investigated the production and role of circulating antibodies, with specificity to staphylococcal enterotoxin B (SEB), in children with AD compared with those of healthy controls. The children with AD had significantly higher levels of serum SEB-specific immunoglobulin G (IgG; p = 0.0193), IgM (p = 0.011), and IgE (p = 0.0001) than the nonatopic children. The proportions of IgG, IgM, and IgE seropositivity in children with AD were 52.5% (21/40), 62.5% (25/40), and 67.5% (27/40), respectively. The levels of SEB-specific IgE and the severity of AD (p = 0.0004) were compared, but no correlation was seen for IgG or IgM. SEB may be involved in exacerbation of AD. SEB-specific IgE may be an important index of the clinical severity of AD. The SEB-specific IgG or IgM was produced during the exposure to the SEB antigen but may not be protective against SEB in AD. PMID- 12635579 TI - The effect of inhaled heparin on airway responsiveness to histamine and leukotriene D4. AB - Inhaled heparin has been shown to reduce the early and late phase of asthmatic reactions and suppress an allergen-induced increase in bronchial hyperreactivity. The mechanism involved in the control of bronchial hyperreactivity in asthma by heparin is still not understood. The purpose of this study was to investigate the effect of inhaled heparin on the airway response to histamine and leukotriene D4. Children with a typical history of mild allergic asthma participated in this randomized, double-blind, placebo-controlled cross-over study. Subjects underwent provocation challenge tests with histamine or leukotriene D4 before and after inhalation of heparin and placebo. Twenty-three patients completed the study. We showed that placebo did not affect the bronchial hyperreactivity to histamine or leukotriene. A single dose of inhaled heparin significantly decreased bronchial hyperreactivity to histamine and leukotriene in children with mild asthma. Results of our study suggest that inhaled heparin, because of its antiallergic and/or anti-inflammatory properties, modifies airway hyperresponsiveness in children with allergic asthma. PMID- 12635582 TI - Is malpractice risk higher for employed physicians? PMID- 12635581 TI - Immediate systemic hypersensitivity reaction associated with topical application of Australian tea tree oil. AB - Australian tea tree oil has been used as a veterinary antiseptic for many years and, more recently, has been extended into human use. There have been many reports of allergic contact dermatitis and toxicity reactions, but it has never been implicated in immediate systemic hypersensitivity reactions. A 38-year-old man experienced immediate flushing, pruritus, throat constriction, and lightheadedness after topical application of tea tree oil. Our purpose was to determine whether this represented an immunoglobulin E (IgE)--mediated reaction. Skin-prick and intradermal testing was performed, as well as enzyme-linked immunosorbent assays for specific IgG and IgE against tea tree oil. The patient had a positive wheal and flare reaction on intradermal testing with tea tree oil. All five patient controls were negative on skin testing. No specific IgG or IgE was detected. We present the first reported case of an immediate systemic hypersensitivity reaction occurring after topical application of Australian tea tree oil, confirmed by positive wheal and flare reaction on skin testing. PMID- 12635583 TI - Coding hospital admissions from other sites of service. PMID- 12635584 TI - Bold initiatives could transform health care. PMID- 12635585 TI - The HIPAA privacy rule: three key forms. Complying with the HIPAA privacy rule may seem trickier than pulling a rabbit out of a hat, but these forms should help. PMID- 12635586 TI - Develop your own practice intranet. It really can be a do-it-yourself project- and you probably have the basic tools already. PMID- 12635587 TI - Putting measurement into practice with a clinical instrument panel. A few key measures can help you gauge whether your practice is headed in the right direction. PMID- 12635588 TI - Ideas for optimizing your nursing staff. PMID- 12635590 TI - HIV poses challenges for Southern states. Most people with AIDS in South are minorities. AB - Southern states have the most HIV/AIDS patients and often are least well-equipped to handle the problem. They also are challenged by an epidemic that is disproportionately African-American and that largely affects the poor. PMID- 12635589 TI - Guarding HIV-positives against vaccine reaction in the age of bioterror. AB - The Bush administration is emphasizing immunization as a way to prevent the U.S. population from being harmed by bioterrorism, especially where smallpox is concerned. But immunology and public health experts are warning that a policy of mass pre-emptive immunization for smallpox would very likely cause harm to HIV positive people, as well as other groups who are at risk for harm from the smallpox vaccine. PMID- 12635591 TI - AIDS destroying hands that rock the world's cradle. HIV infection of women in rich nations also on rise. AB - The latest AIDS epidemic news shows strong evidence that women increasingly are infected with HIV and are dying of AIDS. For the first time, half of the people living with AIDS globally are women, and in sub-Saharan Africa, 60% are women. PMID- 12635592 TI - African prevention efforts yield hope for future. AB - Prevention continues to offer the best hope for stopping the AIDS epidemic, and recent success stories in South Africa and Uganda prove that prevention works, according to a recent report. PMID- 12635593 TI - Dallas group builds a black MSM community. AB - For an AIDS organization to succeed in attracting at-risk populations, it must demonstrate that it cares about the clients and accepts them as they are, says a long-time HIV/AIDS activist who has succeeded where others have failed. Renaissance III is a community center for young African-American men who have sex with men, and it is working. PMID- 12635594 TI - Good advice for immune-compromised patients. No pre-emptive vaccination for HIV positives. AB - Many uncertainties exist concerning how mass inoculations of the smallpox vaccine could affect HIV and AIDS patients. But here's a quick look at the advice many experts are giving. PMID- 12635596 TI - Alternatives to male condoms for men who have sex with men. PMID- 12635595 TI - HIV and syphilis coinfection: trends and interactions. PMID- 12635597 TI - Female-controlled HIV prevention methods. PMID- 12635598 TI - Five new drugs enter the homestretch. PMID- 12635599 TI - First wave of cuts hits New York's ADAP. PMID- 12635600 TI - Interview with Joep Lange. PMID- 12635601 TI - Global treatment update. PMID- 12635602 TI - OraSure begins shipping rapid HIV test to hospitals. CLIA waiver could come in several months. AB - Long desired and long delayed, the first truly rapid HIV test is now available in many U.S. hospitals. The simple, 20-minute test, which uses fingerstick whole blood, was categorized as one of moderate complexity under the Clinical Laboratory Improvement Amendment. However, the tests aren't approved for use in the field where they are needed most. That soon may change if the FDA adopts a waiver requested by the manufacturer. PMID- 12635603 TI - OraQuick counseling advice. AB - Here are key factors to keep in mind in determining how you counsel patients about the new rapid HIV test. PMID- 12635604 TI - CLIA waiver crucial to rapid test adoption. Study finds numerous errors for waived tests. AB - A California study has found that more than half of new HIV infections came from sites that would not be able to use the OraQuick HIV rapid test as it is currently labeled. PMID- 12635605 TI - HIV prevention summit seeks new strategies. Changes in evaluation, reimbursement discussed. AB - HIV prevention is facing a crisis in burnout, complacency, and rising rates of infection threaten health gains made during the past decade. The CDC invited scores of experts to Atlanta recently to brainstorm on how to meet the needs for new generation. PMID- 12635606 TI - Five different audits look at CDC's HIV programs. Critics say they are distracting. AB - Here's a brief look at five different audits the CDC currently is undergoing. Four are being conducted by the Office of the Inspector General, and the other is a self-audit. PMID- 12635607 TI - Prevention strategies abound during CDC summit. Knowledge of serostatus covered. AB - A recent summit at the CDC in Atlanta on HIV prevention strategies included a wide-ranging list of possible ways to deal with knowledge of serostatus. PMID- 12635608 TI - New CDC condom fact sheet invites criticism. What role did politics play in changes? AB - Politics and science are clashing over the facts about condoms and what clinicians should tell their patients about their effectiveness. The CDC's updated condom fact sheet puts more emphasis on abstinence and condom failure. A group of Democratic lawmakers sent a letter to the secretary of Health and Human Services accusing the administration of 'playing politics'. PMID- 12635609 TI - Attitude is key part of treatment adherence. Study suggests which attitudes to watch. AB - A small Chicago study suggests that attitudes about HIV and AIDS among HIV infected people can be broken down into types that predict how well patients will adhere to medication regimens. PMID- 12635610 TI - FDA notifications. New labeling for N-9 products? PMID- 12635611 TI - Bush disavows 'gay plague' remark. PMID- 12635612 TI - Bush surprises advocates with emergency AIDS relief plan. PMID- 12635613 TI - Health care leaders provide Bush AIDS policy suggestions. PMID- 12635614 TI - Disability. Pain, fatigue, opportunistic infections insufficient for SSI. PMID- 12635615 TI - Policy shift. Bush's AIDS plan includes condom distribution, generic drugs. PMID- 12635616 TI - Black Caucus wants AIDS initiative funded immediately. PMID- 12635619 TI - Fear of exposure to HIV insufficient in reused needle case. PMID- 12635617 TI - Spread of the AIDS pandemic. PMID- 12635618 TI - $250,000 award returned to patient for HIV disclosure claim. PMID- 12635620 TI - Health plans. Consumer guide now available for handling health plan disputes. PMID- 12635622 TI - U.S. official to lead Global Fund. PMID- 12635621 TI - Transmission. Failure to disclose serostatus increasingly leads to lawsuits. PMID- 12635623 TI - South Korean to lead WHO. PMID- 12635624 TI - ADAP and Medicaid financial emergencies growing. PMID- 12635625 TI - Smallpox vaccination followup: IOM suggests changes, widespread civilian vaccinations to begin. PMID- 12635626 TI - [What is the procedure in laryngectomy with unilateral radical neck dissection?]. PMID- 12635627 TI - [What is the Tulio phenomenon?]. PMID- 12635628 TI - [Surgical techniques. The ear area]. PMID- 12635629 TI - Medicare outpatient prospective payment system. AB - More and more services that once required hospital stays now are provided on an outpatient basis. However, recent changes to the Medicare payment system for these services have left consumers and professionals confused about how much they should be paying for outpatient procedures. In this brief, we explore the Outpatient Prospective Payment System, including which services are covered (and not covered) and how the costs to your clients are calculated. PMID- 12635630 TI - First Asia-Pacific forum on andrology. 17-21 Oct 2002, Shanghai, China. Proceedings and programme. PMID- 12635631 TI - Manipulation of the cervical spine. AB - In New Zealand, a new approach to manual therapy of the cervical spine has integrated physiotherapy and osteopathy techniques. The combination of the philosophies of these two professions has added a new dimension to the management of cervical spine pain. Emphasis is placed on issues of safety, such as the degree of cervical rotation and comfort for both the patient and the therapist. This is combined with biomechanical considerations, which have made the teaching and learning of these manipulative techniques less complicated and easily progressed from palpation to mobilization and onto manipulation. Appropriate patient screening and selection identified through thorough subjective and objective assessments are important aspects of this approach and reflective interpretation of all clinical findings is essential. The refinement of cervical joint positioning and an increased anatomical awareness have led to the utilization of new upper cervical high-velocity thrust techniques. Consequently, it is envisaged that an increase in the safety and specificity of cervical manipulative techniques is achieved. PMID- 12635632 TI - Foreign-graduate nurses. PMID- 12635634 TI - A call to action on UI. PMID- 12635635 TI - Comparison between color duplex ultrasonography and histology of the temporal artery in cranial arteritis. PMID- 12635636 TI - The effect of postoperative ataralgesia by manual therapy after pulmonary resection. AB - Muscle therapy, a form of manual therapy, was applied to control pain persisting for more than 1 week following posterolateral thoracotomy, and its efficacy for the alleviation of pain was investigated. Eight patients who underwent posterolateral thoracotomy and lung resection for cancer (n=7) or emphysema (n=1) received manual therapy to incised muscles and the muscles inserting into the ribs in the affected area for an average of 17 days postoperatively. Pressure friction and stretching techniques were used. Treatment was continued until the intensity of the pressure-friction technique reached a level at which the patient complained of pain and a decrease in muscle tone was detected. Treatment was performed once a week for 3 weeks. Pain severity was measured using a visual analog scale (VAS) (0-10). Before the first treatment, the VAS was set at 10, and changes of the score were observed before and after the treatment as well as over time. After three sessions, all patients showed a decrease in pain from 10 to an average of 1.9 (range 1.3-2.6). PMID- 12635637 TI - Spinal manipulation for low-back pain: a treatment package agreed to by the UK chiropractic, osteopathy and physiotherapy professional associations. AB - Trials of manipulative treatment have been compromised by, amongst other things, different definitions of the therapeutic procedures involved. This paper describes a spinal manipulation package agreed by the UK professional bodies that represent chiropractors, osteopaths and physiotherapists. It was devised for use in the UK Back pain Exercise And Manipulation (UK BEAM) trial--a national study of physical treatments in primary care funded by the Medical Research Council and the National Health Service Research and Development Programme. Although systematic reviews have reported some beneficial effects of spinal manipulation for low-back pain, due to the limited methodological quality of primary studies and difficulties in defining manipulation, important questions have remained unanswered. The UK BEAM trial was designed to answer some of those questions. Early in the design of the trial, it was acknowledged that the spinal manipulation treatment regimes provided by practitioners from the three professions shared more similarities than differences. Because the trial design specifically precluded comparison of the effect between the professions, it was necessary to devise a homogenous package representative of, and acceptable to, all three. The resulting package is 'pragmatic', in that it represents what happens to most people undergoing manipulation, and 'explanatory' in that it excludes discipline-specific variations and other ancillary treatments. PMID- 12635638 TI - Control subjects in whiplash studies. PMID- 12635641 TI - Changing UI practice. PMID- 12635640 TI - Foodborne diseases. PMID- 12635642 TI - Myocardial Protection from Surgical Ischemic-Reperfusion Injury. Proceedings and abstracts of the 3rd International Symposium. Asheville, North Carolina, USA. June 2-6, 2002. PMID- 12635644 TI - The hidden truths of the belly: the uncertainties of pregnancy in early modern Europe. (Society for the Social History of Medicine Student Prize Essay 1999, runner-up.). AB - For early modern men and women and their medical practitioners, the experience and understanding of pregnancy was primarily uncertain. This uncertainty extended to the whole process of pregnancy--from the moment of conception to delivery, the detection and bearing of a 'true fruit' was doubtful. This 'uncertainty' was heightened by the fact that both body and language could conceal the truth. The woman herself was frequently uncertain and could be mistaken in her interpretation of the condition of her belly. This ambiguity is expressed in the vague and faltering language used to describe such experiences. Women's bodies were believed to conceal the truth more readily than their male counterparts. Equally a woman's physical narrative was more likely to be distrusted. Tensions surrounding the appropriate nature of women's 'knowledge' of such hidden 'secrets' also affected the ways in which women and their practitioners described the 'truths' of the belly. This article traces the ambiguities faced by women and their midwives/accoucheurs through three areas of pregnancy: quickening, false conceptions, and the threat of miscarriage. The much-neglected source of medical texts and observations is drawn upon, alongside letters and diaries and judicial material. PMID- 12635643 TI - Solitary radial head metastasis with wrist drop: a rare presentation of metastatic prostate cancer. AB - The fact that a prostate carcinoma metastasizes to the skeleton is well known. The metastatic deposits are generally multiple and osteoblastic. Moreover, these involve the axial skeleton or the proximal part of long bones. A pure osteolytic metastasis from a prostate carcinoma is extremely rare. This is believed to be the 1st case report of a osteolytic solitary radial head metastasis from a prostate carcinoma in a 60-year-old male. The rare clinical presentation of wrist drop, the absence of metastases at any other site, and a significant decrease in the size of lesion on follow-up in response to hormonal therapy are of interest in this case. The rare clinical presentation of a common malignancy (prostate carcinoma) is reported with a brief review of the literature. PMID- 12635645 TI - Ring-enhanced malignant meningioma mimicking a brain metastasis from a renal cell carcinoma. AB - We report a case of ring-enhanced malignant meningioma mimicking a solitary brain metastasis in a patient with renal cell carcinoma. This misleading situation is rarely encountered and has not been documented previously. Considering the low incidence of brain metastases from a renal cell carcinoma staged T1-2 without systemic metastases, a clinical diagnosis requires circumspection, and both primary and metastatic tumors should be considered when a solitary brain lesion is encountered. PMID- 12635646 TI - Medical advisors for commercial adventures: coming of age? PMID- 12635647 TI - Curing the colonizers: highland hydrotherapy in Guadeloupe. AB - This article explores attempts by French colonial doctors in Guadeloupe to treat tropical pathologies with water and altitude cures, between the seventeenth and twentieth centuries. Assuming that high morbidity rates in the Caribbean were caused by the area's inherent 'toxicity', Europeans turned to mineral water and high-altitude spas, both for the reinvigoration of settlers and to 'season' new arrivals. Guadeloupean spas, though utilized by indigenous, black, creole, and white Guadeloupeans alike, soon emerged as replicas of France in the tropics, and served the socio-medical function of immersing the colonial body in familiar waters. The French obsession with 'healthful space' led to the creation of an 'administrative' quarter precisely in the region of Guadeloupe's spas around Basse- Terre (actually the highest part of the island). The arrival of germ theory in the nineteenth century did not fundamentally alter the rationale for water and altitude cures in Guadeloupe. Tracing the history of these 'folk-cures' -often admonished by colonial doctors as 'common sense', but in reality an integral part of the tropical medical canon--sheds light on some of the underlying dynamics, tensions, and continuities of French imperial medicine. PMID- 12635648 TI - 2002 William Allen Award address. Introductory speech for Albert de la Chapelle. PMID- 12635649 TI - 2002 ASHG award for excellence in education. Introductory speech for Kurt Hirschorn. PMID- 12635650 TI - 2002 Curt Stern Award address. Introductory speech for James R. Lupski. PMID- 12635651 TI - Targeted immunotherapy of cancer: development of antibody-induced cellular immunity. AB - Although immunotherapy of cancer encompasses a large variety of distinct protocols, virtually all therapeutic strategies require the enabling/training of the immune system to distinguish tumour tissue from healthy tissue. In the case of antibody-based therapies, specificity obviously arises from the selectivity of the antibodies for tumour antigens, and tumour cell death derives from either direct cytotoxicity of the antibody or antibody-dependent cellular cytotoxicity. However, even when both of the above killing mechanisms are simultaneously active, we suggest that antibody-based immunotherapies may fall far short of their full potential. In this editorial, we first summarize the mechanisms by which current antibody-based therapies mediate cancer cell removal, and then propose two strategies by which this class of immunotherapies might be further improved. These suggested improvements involve the decoration of tumour cell surfaces with foreign haptens against which an endogenous humoral immune response can be mounted and the recruitment of the cellular arm of the immune system in an antibody-dependent process. PMID- 12635652 TI - Bibliography. Current world literature. Lipid metabolism and therapy. PMID- 12635653 TI - Bibliography. Current world literature. Nutrition in the intensive care unit. PMID- 12635654 TI - Transplantation in Hematology and Oncology III: Stem cell potential, cellular therapy, clinical perspectives. Angiogenesis in Oncology and Hematology II: From basic science to clinical studies. September 6-8, 2002, Munster, Germany. Proceedings and abstracts. PMID- 12635655 TI - Non-steroidal anti-inflammatory drug, nabumetone, prevents indometacin-induced gastric damage via inhibition of neutrophil functions. AB - Nabumetone is a non-steroidal anti-inflammatory drug (NSAID). It works as a prodrug and is extensively metabolized to an active metabolite, 6-methoxy-2 naphthylacetic acid (6MNA). It is well known that neutrophil infiltration and activation are critical in the pathogenesis of NSAID-induced gastric injury, and nabumetone shows less incidence of gastrointestinal irritancy. We examined the effects of nabumetone on neutrophil activation and on indometacin-induced gastric damage. In the indometacin-induced gastric mucosal injury, rats were treated with indometacin and then nabumetone or 6MNA was orally administered. Nabumetone prevented gastric damage accompanied by the reduction of neutrophil infiltration into gastric mucosa, but such an effect was not observed with 6MNA. Nabumetone reduced the formyl methionyl leucyl phenylalanine (fMLP)-induced respiratory burst of human neutrophils to 30% of the control level in-vitro, but 6MNA did not. In addition, nabumetone prevented the fMLP-induced migration of neutrophils. Nabumetone did not inhibit O2- generation in the xanthine-xanthine oxidase system. These results suggest that nabumetone prevents gastric damage induced by the active metabolite, 6MNA, via the suppression of neutrophil activation in gastric mucosa. PMID- 12635657 TI - Chemosensitivity of glioblastoma cells during treatment with the organo-tin compound triethyltin(IV)lupinylsulfide hydrochloride. AB - Malignant gliomas are the most common primary brain tumors in humans. However, poor response to conventional therapeutic approaches, including chemotherapy, leads invariably to disease recurrence and progression. The organo-tin derivative triethyltin(IV)lupinylsulfide hydrochloride (IST-FS 29) was identified and developed as potential antiproliferative agent in human cancer cell lines. However, for its peculiar chemical structure and good lipophilicity, this compound also appeared an eligible candidate for the treatment of gliobastoma cells. The present experiments were designed to explore the in vitro effects of IST-FS 29 on four human glioblastoma cell lines: A-172, DBTRG.05MG, U-87MG and CAS-1. The average IC50 values were obtained by MTT assay and ranged between 3 and 10 microM. Time-course assays with cell recovery after drug withdrawal, demonstrated marked cytotoxicity following exposure to IST-FS 29 for 8, 24 and 72 h. Cultures treated for 8 h were able to partially re-grow by 144 h; on the contrary, longer times of exposure did not allow surviving cells to recover from the damage and actively proliferate. Cell morphology of cultures exposed to IST FS 29 was assessed by inverted light microscopy after 24 and 72 h and was more consistent with cell death by necrosis which included cell size reduction, vacuolation of cytoplasm, round dying cells. The present results and our previous data, in vitro and in vivo, indicate the relevant cytotoxic activity of this organo-tin compound and suggest that IST-FS 29 might be a promising novel agent to be developed for the treatment of malignant brain neoplasms. PMID- 12635656 TI - Involvement of Ras in survival responsiveness to nitric oxide toxicity in pheochromocytoma cells. AB - Nitric oxide (NO) plays a key role in attenuation of tumor growth by activated macrophages that generate large amount of cytotoxic/cytostatic free radicals. However, some tumor cells may survive from NO cytotoxicity and continue to proliferate to malignant tumors. Since a protooncogene product Ras was shown to be activated by NO, this study investigated the involvement of Ras in the cell survival in response to NO cytotoxicity in pheochromocytoma (PC12) cells. Treatment with Ras inhibitor or constitutive expression of dominant negative Ras markedly increased NO-induced cell death. NO-resistant PC12 cells (PC12-NO-R) exhibited higher steady state Ras activity than the parental PC12 cells. Inducible expression using tetracycline-on (Tet-on) system of Ras mutants (dominant negative Ras or dominant active Ras) demonstrated that blockade of Ras activity increased NO-induced cell death whereas enhancement of Ras activity attenuated NO-induced cell death. Furthermore, inducible expression of NO insensitive mutant Ras selectively increased cellular vulnerability to NO but not to ROS. NO, Ras inhibitor and extracellular signal-regulated kinase (Erk) blocker synergistically increased cell death. These observations suggest that Ras activity may be a critical factor for survival response of tumor cells to NO toxicity and pharmacological agents affecting Ras activity may enhance efficacy of NO-mediated tumor therapies. PMID- 12635658 TI - Glioblastoma multiforme in an Asian population: evidence for a distinct genetic pathway. AB - In Singapore astrocytic tumours occur in only 25% of patients with primary brain tumours compared to 40-60% in other series. Glioblastoma multiforme arises either de novo as a primary glioblastomas associated with epidermal growth factor receptor (EGFR) and mdm2 over-expression or as a secondary glioblastomas, through malignant progression from low-grade astrocytomas, associated with p53 mutations and PDGFR-alpha over-expression. Using immunohistochemical methods and DNA sequencing, we studied our population of glioblastomas for overexpression of EGFR, mdm2, p53, and PDGFR-alpha as well directly for mutations of the p53 gene. While levels of over-expression of EGFR and mdm2 were consistent with levels expected for primary glioblastomas, levels of p53 and PDGFR-alpha were consistent with levels documented for secondary glioblastomas. Notably 96% of the samples over-expressed p53 as detected with monoclonal antibody pAb 240. Of the 39 samples available for DNA sequencing 18% (7/39) had p53 mutations, including three mutations previously undocumented in glioblastomas. These results provide strong evidence that glioblastomas in Asian patients do not conform to currently accepted models of glioblastoma development, and that clinically defined glioblastomas in these patients show genetic changes consistent with both 'primary' and 'secondary' glioblastomas. PMID- 12635660 TI - Ceramide increases Fas-mediated apoptosis in glioblastoma cells through FLIP down regulation. AB - Ceramide is a physiologic regulator of growth and differentiation in mammalian cells. In this study, the relationship between ceramide and FLICE inhibitory protein (FLIP) in the induction of apoptosis in glioblastoma cell lines was investigated. We found that LN215 cells were slightly more sensitive to Fas mediated apoptosis than LN319 cells, which were more sensitive to ceramide than LN215 cells. FLIP was expressed in LN319 and LN215 cells constitutively, and this expression decreased with treatment of ceramide in LN215 cells, which might cause LN215 cells to be more sensitive to Fas-mediated apoptosis at lower level stimulation. In LN319 cells FLIP levels were not modified by ceramide treatment and the level of cell death induced by anti-Fas antibody was not affected. Our results suggest that FLIP may be down-regulated by low levels of ceramide in LN215 cells, which causes LN215 cells to be more sensitive to Fas-mediated apoptosis, whereas LN319 cells remain resistant. PMID- 12635659 TI - CD44H is expressed by cells of the oligodendrocyte lineage and by oligodendrogliomas in humans. AB - CD44, a family of cell surface glycoproteins involved in cell-cell and cell extracellular matrix adhesion, is widely expressed in the white matter of the normal brain and in astrocytic gliomas under its standard form (CD44s also called CD44H). On the other hand, several variants have been found in brain metastases and rarely found in gliomas. We have investigated by immunohistochemistry CD44H and CD44v6 expression in 28 oligodendrogliomas. All tumors were CD44v6 negative whereas nearly all tumors were immunolabelled with anti-CD44H antibody. Immunostaining increased in parallel with grade and was particulary strong around vessels and in tumoral subpial nodules. Western blot analysis showed that oligodendrogliomas expressed the same 80-kDa CD44 isoform as normal brain. Since gliomas may arise from a dividing progenitor cell, we also studied CD44H expression during the oligodendrocyte lineage in vitro in parallel with specific markers of the O-2A cells. Precursor cells (PSA-NCAM positive), O-2A progenitor cells, as well as preoligodendrocytes (A2B5 positive cells) and immature oligodendrocytes (O4 positive cells), coexpressed CD44H. Our data showed that CD44H is expressed by cells of the oligodendrocyte lineage in vitro and by oligodendrogliomas in vivo especially in sites of dissemination such as subpial spaces. This suggests that CD44H could play a role in migration of tumor cells in oligodendrocytic tumors. PMID- 12635661 TI - Distribution of adoptively transferred, tumor-sensitized lymphocytes in the glioma-bearing rat. AB - For adoptively transferred lymphocytes to exert anti-tumor effects in vivo, they must traffic or initiate the migration of endogenous immune cells to the site of tumor. Using a rat model, we examined the trafficking of tumor-sensitized lymphocytes to an intracerebral glioma. By labeling the cells with 111Indium oxine (111In) prior to intravenous injection, we were able to quantify the relative number of lymphocytes that traveled to the tumor site. There was no difference in lymphocytic influx between the tumor-bearing and non-tumor-bearing cerebral hemispheres in 3-day rat glioma models. However, in 7-day models, significantly greater numbers of 111In-labeled lymphocytes resided in the tumor bearing hemisphere at 12 h post-administration. This number increased more than two-fold by 24 h post-adoptive transfer. Using fluorescent-labeled lymphocytes and microscopy, we confirmed that the detection of radioactivity within the brain was truly due to tumor infiltrating 111In-labeled lymphocytes. Adoptively transferred cells were found in perivascular and peritumoral locations. These data demonstrate that tumor-sensitized lymphocytes traffic to an intracerebral target site where they can exert an effect, further supporting adoptive immunotherapy as a treatment for glioma. PMID- 12635662 TI - New approach to tumor therapy for inoperable areas of the brain: chronic intraparenchymal drug delivery. AB - Because the brainstem has little functional redundancy, diffuse lesions have been regarded as inoperable. To determine whether local drug therapy can prolong survival in a rodent model of a tumor in such eloquent tissue, lethal doses of F98 and 9L tumor cells were injected into the brainstems of Fischer 344 rats. Five days after inoculations, 0.5 mg/ml solutions of carboplatin were infused at 1 microl/h for 7 days. Compared to control groups that survived 13-17 days with F98 tumors and 22-23 days with 9L tumors, animals locally infused with 0.1 mg of carboplatin survived 27-30 days (Prob > Chi Sq = 0.0003), and 32 days (Prob > Chi Sq = 0.01), respectively. Measurements of tissue platinum levels at autopsy suggested that infusions distributed pharmacologically relevant levels of carboplatin through a volume of tissue at least 0.5 cm in diameter. The results suggest that chronic low-flow infusions provide a promising approach to therapy for CNS lesions in tissues considered to be inoperable. PMID- 12635664 TI - Optic neuropathy in cancer patients. Report of a case possibly related to 5 fluorouracil toxicity and review of the literature. AB - We report the case of a 72-year-old woman suffering from bone metastatic breast cancer and treated with continuous intravenous 5 fluorouracil (5FU) chemotherapy, who presented with a bilateral asymmetric anterior optic neuropathy (ON). A deficiency of dihydropyrimidine dehydrogenase (DPD) was documented. Patients with DPD deficiency are at increased risk for developing unusual and/or severe toxicity to 5FU. We discuss the differential diagnosis of anterior ON and we suggest that 5FU can be responsible for optic nerve toxicity in patients with DPD deficiency. PMID- 12635663 TI - Meningioma-associated brain oedema: the role of angiogenic factors and pial blood supply. AB - BACKGROUND: Approximately 60% of meningiomas are associated with perilesional brain oedema. Several aspects have been evaluated in order to understand the pathophysiological mechanisms of oedema (age, sex of the patient, size and location of the tumour, histotype, grading), although at present they have yet to be completely clarified. We focused on pial blood supply, microvascular density (MVD) and angiogenic growth factors (i.e. vascular endothelial growth factor- VEGF) in order to evaluate their putative role in the development of brain oedema. METHODS: We retrospectively studied 55 patients with intracranial meningiomas. Computerized tomography (CT) and angiographic studies were obtained in all cases. The angiograms provided an accurate differentiation between pial and dural blood supply, concomitantly with its semi-quantitative evaluation. The location and the volume of oedema, in relation to the meningioma surface, was evaluated using CT scans, as an oedema index (E/I). We also determined the expression of VEGF and MVD using standard immunohistochemical methods. RESULTS: Thirty-two out of 55 meningiomas presented peritumoural oedema, with an angiographic blush ranging from 2 to 4; VEGF protein was expressed in 27 out of 32 cases, independent of grade or histotype of tumours. In all patients, MVD ranged from 4 to 33.3 vessels (median value: 10.6). A significant relationship was found between the expression of VEGF and MVD (p = 0.0003) and between VEGF and E/I (p = 0.0023). Moreover, the E/I ratio was related to the blush (p = 0.0005). A significant association was also present between VEGF expression and pial blush (p = 0.0001). CONCLUSION: Our data confirm the central role of VEGF and pial blood supply in the pathogenesis of peritumoural oedema and support the hypothesis that the development of oedema in meningioma is vasogenic in type. PMID- 12635665 TI - Phase II study of chemotherapy with ACNU plus cisplatin followed by cranial irradiation in patients with newly diagnosed glioblastoma multiforme. AB - PURPOSE: To evaluate the efficacy and toxicity of chemotherapy with ACNU (1-(4 amino-2-methyl-5-pyrimidinyl)-methyl-(2-chloroethyl)-3-nitrosourea) plus cisplatin followed by cranial irradiation in patients with newly diagnosed glioblastoma multiforme. PATIENTS AND METHODS: Between August 1999 and July 2001, previously untreated 30 patients with histologically confirmed glioblastoma multiforme were treated. Chemotherapy consisting of up to 2 cycles of 72 h of continuous intravenous infusion of ACNU (40 mg/m2/day) and cisplatin (40 mg/m2/d) was given over a 6-week period. Radiation was begun 6 weeks after the second cycle of chemotherapy. RESULTS: Median age was 48 years (range 18-66 years) and 22 patients with residual measurable disease after surgery were eligible for response analysis. One (5%) had a complete response (CR), 36% partial response (PR), 14% stable disease (SD), and 45% progressive disease (PD) after chemotherapy. After additional radiation, 22% had CR, 22% PR, 16% SD, and 42% PD. Grades III and IV leukopenia and thrombocytopenia occurred in 18 cycles (36%) and 15 cycles (30%), respectively. No fatal complications occurred. Median time to progression was 5.9 months (95% CI 5.1-6.8 months) and median overall survival was 14.9 months (95% CI 9.1-20.7 months). CONCLUSIONS: Preradiation chemotherapy with ACNU plus cisplatin is effective and feasible in patients with gliobiastoma multiforme. PMID- 12635666 TI - Three-dimensional conformal radiotherapy for astrocytic tumors involving the eloquent area in children and young adults. AB - PURPOSE: Although a gross total removal of astrocytic tumors offers a favorable prognosis, it is often difficult to achieve in the eloquent area of the brain. This study was conducted to investigate the possible gain of three-dimensional conformal radiotherapy (3DCRT) for astrocytic tumors located in the eloquent area in children and young adults. MATERIALS AND METHODS: Twenty patients with astrocytic tumors received the radiotherapy. The median age was 17 years, ranging from 4 to 30 years. Fourteen low-grade tumors (seven pilocytic and seven diffuse), and six high-grade tumors (five anaplastic, one malignant pilocytic) were included. Tumors were located at the thalamus/hypothalamus in 12 cases, optic tract in one case, and the deep cerebral/cerebellar hemisphere in seven cases. A specific fixation device was used for 3DCRT. Forty-six Gy for low-grade tumors and 54 Gy for high-grade astrocytomas with 1.8-2.0 Gy per fraction were in principle employed as the standard regimen. Nominal radiotherapy fields ranged from 2.0 x 2.0 to 15.0 x 11.0 cm2. The median follow-up period was 42 months, ranging from 3 to 108 months. RESULTS: The actuarial survival rate at 5 years was 68% +/- 13% for all patients. The actuarial survival rate for low-grade glioma was 79% +/- 14% at 5 years and 50% +/- 20% at 3 years for high-grade glioma. The actual progression-free survival rate was 83% +/- 11% at 5 years for low-grade glioma and 50% +/- 20% for high-grade glioma. A complete response was obtained in three (21%) of 14 patients with low-grade astrocytic tumors. Two patients with low-grade tumors and four of six with high-grade tumors died due to tumor progression with infield relapse but not marginal relapse. Twelve survivors with low-grade tumors showed no signs of relapse and no neurological, hormonal, or cognitive deterioration after radiotherapy and were able to attend their school or continue with a full-time job. CONCLUSIONS: 3DCRT is safe and effective for low-grade astrocytic tumors located in the eloquent area in children and young adults. PMID- 12635667 TI - Surgical treatment of pituitary tumors in the elderly: clinical outcome and long term follow-up. AB - Pituitary adenomas in the elderly represent an increasing proportion of pituitary tumors, making the surgical management of these patients of special importance. We therefore decided to review our experience with transsphenoidal surgery (TSS) in this age group, in order to better evaluate its efficacy, safety and outcome. A retrospective study was performed on 39 patients aged more than 70 years at the time of surgery (mean age 74.1 +/- 2.9 years) and with a minimum follow-up duration of 4 years at the time of the study. Thirty-one had a nonfunctioning pituitary adenoma, 5 had a GH-secreting adenoma and 3 a macroprolactinoma, respectively. The commonest presenting symptom was visual deterioration (60%), whereas hypopituitarism were present in 41% of patients. All patients underwent TSS without any major peri- and post-operative complications. Visual fields improved in 74% of patients. Post-operative radiotherapy was performed in 17 patients with partial surgical resection and/or persistent hormonal hypersecretion, whereas 22 patients with subtotal or total surgical resection were treated by surgery only. Long-term follow-up (mean 9.1 +/- 2.7 years) showed evidence for tumor regrowth in one irradiated (5.9%) and 3 non-irradiated patients (13.6%), respectively. A noticeable complication of radiotherapy was progressive hypopituitarism, which worsened or developed in 65% of patients. We conclude that TSS is safe and well tolerated even in elderly patients, with a low incidence of minor complications. The indications for post-operative radiotherapy in patients with incomplete tumor resection should be better defined. PMID- 12635668 TI - Apoptosis and lens vesicle development. AB - PURPOSE: To study the development of the rat lens vesicle in relation to apoptosis. MATERIALS AND METHODS: Fetuses of Wistar Kyoto rats were removed by laparotomy on day 10-15 of gestation. Some fetuses were fixed in 2% paraformaldehyde and embedded in paraffin for a TUNEL technique examination of DNA fragmentation. Macrophages were stained immunohistochemically with antibody. Some fetuses were fixed in 4% glutaraldehyde and 1% osmic acid and embedded in Luveak 812, then examined with a transmission electron microscope (TEM). RESULTS: On day 11 of gestation (E11) before the start of lens invagination, apoptotic changes were noted in the cells between the surface ectoderm and optic vesicle, with the appearance of phagocytic cells. Apoptotic cells were present at the junction of the surface ectoderm and the lens placode, in the ventral and dorsal thirds of the lens placode and in the outer layer of the optic vesicle in the same axes on E12. Apoptotic changes appeared in the lens stalk, surface ectoderm and the anterior lens epithelium on E12.5. The lens vesicle was detached completely from the surface ectoderm by E13 and some cells had the typical characteristics of macrophages in the extracellular space between the surface ectoderm and the anterior lens epithelium. Apoptotic changes were confirmed by the TUNEL method, and macrophages were stained immunohistochemically. CONCLUSIONS: Apoptosis may have a major role during the whole process of lens vesicle development. Apoptosis may eliminate the cells between the surface ectoderm and the optic vesicle, help trigger invagination and facilitate separation from the ectoderm. Apoptosis might aid in the bowing of the optic vesicle during lens invagination. PMID- 12635670 TI - Ocular hypotensive effects of cholinergic and adrenergic drugs may be influenced by prostaglandins E2 in the human and rabbit eye. AB - PURPOSE: Increased PGE2 production by the iris and ciliary body regulate intraocular pressure (IOP) in vivo. Various cholinergic and adrenergic compounds are traditionally used as antiglaucoma drugs, and their effect on IOP reduction is antagonised by cyclooxygenase inhibitors, indicating a role for eicosanoids in their hypotensive activity. One of the most potent antiglaucoma drugs, PG2 alpha (Latanoprost), reduces IOP by increasing uveoscleral outflow and also increases PGE2 production by the iris and ciliary body in vivo. We investigated whether cholinergic and adrenergic antiglaucoma drugs induce the production of prostaglandin E2 (PGE2) in vitro by: 1) the iris-ciliary body (ICB) of rabbits and, 2) irises of glaucoma patients. METHODS: Pilocarpine 2%, epinephrine 1% and echothiophate iodide 0.125% were applied topically to both eyes of Albino rabbits. Control groups were treated with the corresponding vehicles, or untreated completely. Human iris specimens were obtained from nine untreated cataract eyes, and five eyes under antiglaucoma medication undergoing surgery. PGE2 were determined by a radioimmunoassay. RESULTS: PGE2 production by the ICB of treated rabbits in vitro was twice that of vehicle-treated or untreated rabbit eyes (p<0.001, for either group). In vitro PGE2 production by treated glaucoma patients' irises was three times higher (p<0.001) than in cataract control patients. CONCLUSIONS: The study found an increase in in vitro production of PGE2 by the irises of eyes treated with cholinergic and adrenergic antiglaucoma medications. This suggests a role for endogenous PG production in the hypotensive effect of both classes of drug. PMID- 12635669 TI - Does identification of the causal organism of corneal ulcers influence the outcome? AB - PURPOSE: To investigate whether identification of the causal organism in corneal ulcers influences their outcome. METHODS: We retrospectively studied 114 patients, 72 males and 42 females aged 6-89 years, admitted to this eye clinic during the years 1994-2000 on account of an infectious corneal ulcer. Their examination included a detailed history, visual acuity measurement, and biomicroscopy in everyday follow-up. The ulcers were classified according to their severity and outcome. We assessed the cases where cultures had been done, reviewed the results, and searched for a possible correlation between the outcome and the fact of culturing the ulcer and identifying the causal organism. RESULTS: Of the 114 corneal ulcers studied, 23 were mild, 49 moderate, and 42 severe. Fifty (44%) had not been cultured, but 64 ulcers (56%) had been cultured, with a positive result in 37 cases (58%), Staphylococcus and Pseudomonas species being the most common organisms found. In moderate and severe ulcers, there was a tendency to a higher proportion of successful outcome for cultured ulcers, but with no significant correlation. CONCLUSIONS: Despite a tendency towards favorable results in culture-positive corneal ulcers, the influence of the detection of the organism on their outcome has not been proved. The role of the initial broad-spectrum antibiotic therapy remains important. PMID- 12635671 TI - Comparative analysis of the effects of dorzolamide and latanoprost on ocular hemodynamics in normal tension glaucoma patients. AB - PURPOSE: To compare the effects of latanoprost (Xalatan) and dorzolamide (Trusopt) on ocular hemodynamics in normal-tension glaucoma patients. METHODS: A randomized, single-masked, parallel design study was conducted. After a 4-week washout period, 20 normal tension glaucoma patients, recruited from a single university-based ophthalmology clinic, received either latanoprost once daily or dorzolamide 3 times daily for 4 weeks. The subjects were examined at baseline and post-treatment. Outcome measures included heart rate (HR), blood pressure (BP), logMar visual acuity (VA), contrast sensitivity (CS), intraocular pressure (IOP), color Doppler imaging (CDI), and fluorescein angiography with the Rodenstock scanning laser ophthalmoscope (SLO). CDI measurements of the retrobulbar vessels included peak systolic velocity, end diastolic velocity, and the calculated resistance index. Arterio-venous passage time (AVP) in the superior and inferior temporal retina was calculated from the SLO angiograms. RESULTS: Neither dorzolamide nor latanoprost had any statistically significantly effect on HR or BP. Both drugs significantly lowered IOP without altering calculated ocular perfusion pressure (p<0.05). There was no statistically significant difference in any CDI measurement. Dorzolamide significantly decreased AVP time in the superior retina (p=0.011), while latanoprost did not (p=0.62). CONCLUSIONS: Dorzolamide, unlike latanoprost, significantly reduced AVP times in the superior temporal retina in normal tension glaucoma (NTG) patients. PMID- 12635673 TI - Determination of optic cup depth by confocal scanning laser tomography. AB - PURPOSE: To assess whether confocal scanning laser tomography of the optic disc and clinical ophthalmoscopy using stereoscopic optic disc photographs, agree in the evaluation of the deepest part of the optic cup. PATIENTS AND METHODS: The study included 33 eyes of patients with focal normal-pressure glaucoma. Using 15 degrees color stereoscopic optic disc photographs and dividing the optic disc into an upper and a lower half, two trained observers independently established the location of the deepest part of the cup. By selection, the two observers had made congruent judgements for all eyes in the study. All patients also underwent confocal scanning laser tomography of the optic disc using the Heidelberg Retina Tomograph (HRT). The location of the deepest "mean cup depth" was noted. RESULTS: The deepest optic cup region based on stereoscopic evaluation of photographs and on scanning laser tomography did not correlate significantly with each other (p=0.18; chi-square test, linear-by-linear association). According to the scanning laser tomography data, the maximal optic cup depths were not correlated with the corresponding locations of maximal visual field defect (p=0.80). Using the stereo photographic data, the locations of maximal optic cup depth and maximal visual field defect were correlated (p=0.006). CONCLUSIONS: In some glaucomatous eyes, confocal scanning laser tomographic assessment of the location of the deepest optic cup region does not agree with clinical judgement and, in contrast to clinical examination, does not correlate with the location of the maximal visual field defect. PMID- 12635674 TI - A method for detecting progression in glaucoma patients with deep, localized perimetric defects. AB - PURPOSE: To define a method for early detection of progressive visual field loss, based on monitoring the "healthy" component of the visual field, in glaucoma patients whose perimetric findings show the co-existence of deep scotomata and normal sensitivity areas. METHODS: We reviewed all the "central 30-2 threshold tests" stored in the oldest of our Humphrey perimeters (a 640 VFA model, in use at the Glaucoma Service of the University Eye Clinic of Genoa since 1986). Only the perimetric findings of glaucoma patients with pure, deep, localized defects were collected for this study. In accordance with several inclusion criteria, we could select only 12 series of consecutive examinations (12 eyes of 12 patients). Each series included 12 to 20 examinations and the observation period ranged from 6 years 2 months to 9 years 4 months. Some pre-defined criteria made it possible to separate the defective component of the visual field from the "healthy" one. Then two independent "mean deviations" were calculated, one related to the "healthy" area and one to the defective one. RESULTS: The mean deviation related to the "healthy" component of the visual field showed very little variation (0.6 to -1.3 dB) in the four series that had no increase in defects, even at the end of the observation period. However, in 7 of the 8 series with a tendency to worsen there was a small inter-test increase (-2.2 to -2.6 dB). This finding anticipated the enlargement of the scotomata, confirmed by subsequent examinations. Only in one series did the increase of the mean deviation related to the "healthy" area coincide in time with the real deterioration of the visual field, rather than anticipating it, but the inter-test interval had by chance been much longer than in the other series. The mean deviation related to the defective areas always showed very large changes in all the series, caused by the high variability of thresholds inside scotomata. This was the explanation for the large variations revealed by the "global" mean deviation too. CONCLUSIONS: Detecting progression is still one of the major problems in evaluating perimetric results. It might be easier to achieve this goal with a method for selectively monitoring light sensitivity inside the "healthy" areas of the visual field. PMID- 12635675 TI - Topical aminocaproic acid to prevent rebleeding in cases of traumatic hyphema. AB - PURPOSE: To determine the effect of topical aminocaproic acid on the incidence of rebleeding after traumatic hyphema. PATIENTS AND METHODS: This randomized double blind clinical trial investigated 132 consecutive cases of traumatic hyphema referred to the emergency room of Farabi Eye Hospital in 1998-1999. The patients were randomly divided into three groups: Group 1 received cycloplegic drops only. Group 2 received cycloplegic drops and 2% carboxy polymethylene (CPM) gel as placebo. Group 3 was treated with cycloplegic drops and 25% aminocaproic acid (ACA) in CPM gel (supplied by Messrs. Sina Darou). All patients were treated for five days on an outpatient basis, with a two-week follow-up. The incidence of rebleeding, time needed for clot absorption, and complications of hyphema were recorded and analyzed using the chi-square and Student's t-tests and logistic regression modeling. RESULTS: Rebleeding occurred in 8 eyes of 52 patients in group 1 (15.4%), 7 eyes of the 39 patients in group 2 (17.9%) and 5 eyes of the 41 patients in group 3 (12.2%). This difference was not significant. The time needed for clot absorption in groups 1, 2 and 3 was respectively 9.5 +/- 3.9, 9.3 +/- 4.2 and 11.15 +/- 4.7 days, the difference between group 3 and the other two groups being statistically significant (p<0.04). CONCLUSIONS: Topical 25% ACA is not effective in reducing the incidence of rebleeding and lengthens the time needed for clot absorption. PMID- 12635672 TI - Multicenter evaluation of tendency-oriented perimetry (TOP) using the G1 grid. AB - PURPOSE: The G1-TOP program is a short automated perimetric strategy which sub divides the G1 grid of 59 points into four sub-grids. Each point is tested only once, but each patient's response is used to modify that particular point and the surrounding ones from the remaining sub-grids. This study compared the results of the G1-TOP program with the Standard Bracketing strategy. METHODS: Eleven participating institutions provided data from 213 patients (406 eyes). The main group consisted of 284 glaucomas and 55 glaucoma suspects. Other groups included 31 eyes with neurological disorders, 20 with chorioretinal lesions and 16 normal eyes. Mean age was 62.7 +/- 15.4 (range 14-88) years. All subjects had previous perimetric experience and visual acuity better than 0.5. Examination included G1 Standard Bracketing and G1-TOP testing, in interchangeable order, with the Octopus 1-2-3 perimeter. RESULTS: The correlation coefficient for mean defect (MD) was 0.95. Standard error (YX) for MD, square root of loss variance (LV) and individual thresholds were 1.86 dB, 1.29 dB, and 4.72 dB, respectively. Mean sensitivity values were similar (difference 0.04 +/- 1.87 dB) (p>0.05). Mean duration for G1-TOP was 2.19 +/- 0.26 min, while G1-Standard Bracketing took 11.51 +/- 1.52 min (ratio 1/5.1, or a net reduction of 80.4%). The sensitivity of G1-TOP versus G1-Standard Bracketing was: glaucoma 77.1/78.5, glaucoma suspects 38.2/47.3, neurological disorders 87.1/87.1 and chorioretinal lesions 80.0/85.0. CONCLUSIONS: The G1-TOP program gave very similar results to G1-Standard Bracketing in only 20% of the time required by the standard strategy. PMID- 12635676 TI - Implantation of a black diaphragm intraocular lens in ten cases of post-traumatic aniridia. AB - PURPOSE: To retrospectively review the safety and efficacy of black-diaphragm intraocular lenses (IOL) implanted for the treatment of post-traumatic aniridia. METHODS: Ten patients (mean age 48 years, range 21-75) were implanted with a black-diaphragm posterior chamber IOL (Morcher GmbH, model 67F) for correction of post-traumatic aniridia associated with cataract or aphakia. This IOL, in poly(methylmethacrylate), consists of an opaque diaphragm surrounding the transparent optic, and was inserted through a 10-mm scleral tunnel (seven eyes) or through the corneal trephination in cases of simultaneous penetrating keratoplasty (three eyes), and in-the-sulcus implanted, trans-sclerally sutured (six eyes) or on capsular support (four eyes). Mean follow-up was 33.4 months (range 12-52). RESULTS: Best-corrected visual acuity (BCVA) improved in eight eyes and remained unchanged in two. Glare and photophobia decreased in all patients. Intraoperatively, ciliary sulcus bleeding occurred in two cases and haptic rupture during lens insertion in one. Postoperatively, persistent intraocular inflammation was seen in four eyes, secondary glaucoma in four eyes, transient hyphema and/or hemovitreous in four, IOL decentration in two, and post traumatic haptic detachment in one eye. CONCLUSIONS: Although in our experience the haptics still seem weak and the diaphragm diameter too large, implantation of the black-diaphragm IOL type 67F appeared sufficiently safe and provided satisfactory functional results for correction of post-traumatic aniridia combined with cataract or aphakia, improving BCVA and reducing glare and photophobia in most patients, though clearly more cases and longer follow-up are needed to assess its clinical performance properly. PMID- 12635677 TI - Vitreoretinal surgery for retained lens fragments after phacoemulsification. AB - PURPOSE: To evaluate the prognostic factors, effect of timing and outcomes of vitreoretinal surgery for removal of retained lens fragments after phacoemulsification. METHODS: A retrospective review of 43 eyes of 43 patients who had vitreoretinal surgery for retained lens fragments after phacoemulsification, between January 1998 and November 2000. RESULTS: Seven of the 43 patients underwent vitrectomy on the same day as cataract surgery, 20 in the first week, and 23 after the first week, with a mean of 14.8 days (0-90). Initial visual acuity was < or = 20/400 in 27 (75%, n=36) and intraocular pressure (IOP) > or = 25 mmHg in 22 (61%, n=36) eyes with or without medication. The mean preoperative IOP was 27.4 mmHg. Initial ocular findings included moderate or severe corneal edema in 17 patients (40%), uveitis in 14 (33%), retinal detachment in 1 (2%) and vitreous hemorrhage in 2 (5%). After a mean follow-up of 8.4 months, final best-corrected visual acuity (BCVA) was > or = 20/40 in 24 patients (56%) and < or = 20/400 in 7 (16%). Persistent corneal edema (one eye), cystoid macular edema (four eyes), age-related macular degeneration (one eye) and suprachoroidal hemorrhage (one eye) were the causes of BCVA < or = 20/400. Final mean IOP was 15.2 mmHg and only one case had IOP > 25 mmHg. Uveitis disappeared in all cases (p<0.001), and corneal edema persisted in only one eye. Both the BCVA and IOP differences were significant (p<0.001), but no correlation was found between pre- and postoperative BCVA and IOP as regards vitreoretinal surgery timing, posterior or anterior removal sites and IOL implantation sites or procedures. CONCLUSIONS: Vitreoretinal surgery is effective for removing retained lens fragments after phacoemulsification, lowering the IOP and reducing the uveitic reaction and corneal edema. BCVA > or = 20/40 can be reached in at least half the patients. PMID- 12635679 TI - Protection of the retina from ischemia-reperfusion injury by L-carnitine in guinea pigs. AB - PURPOSE: To investigate the efficacy of L-carnitine in preventing retinal injury followed by ischemia-reperfusion. METHODS: The eyes of 34 guinea pigs were used in this experiment. The guinea pigs were divided into two groups: the first group (n=17) was given L-carnitine intraperitoneally (500 mg/kg) and second group (n=17) received the same dose of saline solution. Under general anesthesia, peritomy was performed. Retro-orbital tissues were ligated for 90 minutes and ischemia was induced, followed by 4 hours of reperfusion. One of the enucleated eye was stained with hematoxylin and eosin (H&E) and retinal thicknesses were evaluated. Thiobarbituric acid reactive substances (TBARS) levels were determined in the retina of the other eye. RESULTS: Mean TBARS levels in retinal tissue were found lower in L-carnitine group (2.77 +/- 0.55 microM) than in the control group (6.57 +/- 1.19 microM), (p<0.01). On the other hand, mean retinal thickness was found to be increased in the control group (47.47 +/- 5.62 microm) when compared to the L-carnitine group (26.52 +/- 4.65 microm), (p<0.01). In correlation analysis, significantly positive relationships were found between retinal TBARS level and retinal thickness both in the control and L-carnitine groups (r=0.981, p<0.01 and r= 0.967, p<0.01 respectively). CONCLUSIONS: L-carnitine is effective in preventing retinal injury followed by ischemia-reperfusion. PMID- 12635678 TI - Retinochoroiditis associated with congenital toxoplasmosis in children: IgG antibody profiles demonstrating the synthesis of local antibodies. AB - PURPOSE: Retinochoroiditis is generally diagnosed after the first year of life and the association with congenital toxoplasmosis presents a diagnostic dilemma. The detection of local intraocular specific antibodies could be useful for diagnosis. METHODS: We studied six patients (mean age 7 +/- 5 years) with retinochoroiditis which appeared after the first year of life. Aqueous and serum were analysed by immunoblotting for anti T. gondii IgG to diagnose toxoplasmosis. RESULTS: All serum samples were positive only for anti T. gondii IgG. The retinochoroiditis was active in three patients and inactive in the others. Immunoblot analysis of serum and aqueous from the patients with active lesions showed IgG versus the specific antigen of T. gondii. In the patients with inactive lesions the pattern was the same in the two compartments. In active forms, aqueous and serum Western blot patterns differed in proteins lower than 16kDa and higher than 116kDa: in aqueous the findings were typically positive for 30kDa. CONCLUSIONS: Aqueous humour analysis by the Western blot technique may be useful in the diagnosis of congenital toxoplasmosis. In the present small series, we nevertheless detected different patterns for inactive and active retinochoroiditis, confirming the diagnosis in the latter. Aqueous humour paracentesis may be indicated in a child with active retinochoroiditis with unusual clinical features, appearing after the first year of life, and with no clinical or serological evidence of congenital infection. PMID- 12635680 TI - Large subconjunctival emphysema causing diplopia and lagophthalmos. AB - PURPOSE: To describe a patient who developed diplopia, lagophthalmos and exposure keratopathy due to a large subconjunctival emphysema. METHODS: A 24-year-old man sustained an injury in his left eye from a compressed air hose. The patient complained of pain and diplopia. He underwent slit-lamp examination, funduscopy and computed tomography. RESULTS: Ophthalmic examination revealed a decrease in vision in the left eye to 0.5, a conjunctival laceration adjacent to the medial limbus, subconjunctival hemorrhage, a large subconjunctival emphysema, lagophthalmos, hypertropia and superficial punctate keratopathy. The posterior pole was intact as were the orbital bones. Two weeks after the injury the conjunctival emphysema, diplopia, lagophthalmos and superficial keratopathy resolved, and visual acuity improved to 1. CONCLUSIONS: Large subconjunctival emphysema can result in diplopia, lagophthalmos and exposure keratopathy. PMID- 12635681 TI - Corneal neovascularization possibly associated with latanoprost therapy. AB - PURPOSE: To report a case of corneal neovascularization possibly associated with latanoprost therapy. METHODS: CASE REPORT: A 67-year-old man developed a progressive stromal corneal neovascularization in his right eye within eight months of a corneal trauma. At admission, he was receiving latanoprost 0.005% therapy. His topical medications were rearranged: latanoprost was replaced with carteolol hydrochloride 1% twice daily bilaterally and prednisolone acetate 1% was added twice daily in the right eye. RESULTS: One month later, he presented regression of the corneal neovascularization and an increase in visual acuity. CONCLUSIONS: Latanoprost, an arachidonic acid derivative, could have directly or indirectly stimulated the corneal neovascularization in this patient with a history of nonpenetrating corneal trauma. PMID- 12635682 TI - Indirect trauma causing dislocation of the crystalline lens--a case report. AB - PURPOSE: To report a case of indirect trauma causing dislocation of the crystalline lens in a patient with no underlying risk factors. METHODS: Case report. RESULTS: A 76-year-old woman fell injuring her right temple area. She complained of blurring of vision in her right eye. Her vision was reduced to 6/60. Ocular examination revealed right aphakia associated with complete posterior dislocation of the crystalline lens into the vitreous cavity. No predisposing factors as pigment dispersion or pseudoexfoliation material was found on gonioscopy. CONCLUSIONS: Dislocation of the crystalline lens has been reported following direct trauma. It is known to occur spontaneously in mature and hypermature cataracts and in cases of weak zonules. Our case shows that indirect trauma can cause complete dislocation of the crystalline lens in the absence of any predisposing factors. PMID- 12635683 TI - Uveitis associated with granuloma annulare. AB - PURPOSE: To report a case of uveitis associated with granuloma annulare (GA). GA is a benign, usually self-limited dermatosis of unknown origin, causing necrobiotic dermal and subcutaneous papules. So far, ocular involvement has not been reported in GA. METHODS: We describe a case of uveitis with GA. Ocular examination findings were consistent with uveitis. Histopathologic studies on lower extremity lesions revealed features consistent with GA. RESULTS: The patient was successfully treated with systemic and topical corticosteroids. After two months, however, the skin and eye lesions relapsed. When the same treatment was restarted, her lesions regressed within four weeks. CONCLUSIONS: This case provides an example of concomitant uveitis and GA. To our knowledge, this is the first report of this combination. PMID- 12635684 TI - Intraocular hemorrhage after systemic thrombolytic therapy in a patient with exudative macular degeneration. AB - PURPOSE: To report a hemorrhagic complication from thrombolytic therapy in a patient with exudative macular degeneration. CASE REPORT: A 75 year old patient with exudative macular degeneration developed pain and loss of vision in the left eye shortly after receiving tissue plasminogen activator (t-PA) for a myocardial infarction. Examination revealed the patient to be in angle closure. A CT scan revealed the etiology of the angle closure to be a dense vitreous hemorrhage pushing the iris-lens diaphragm forward. Intraocular pressure was treated successfully, but the final visual acuity was only light perception. CONCLUSIONS: Thrombolytic therapy can lead to devastating intraocular hemorrhages. The presence of exudative macular degeneration may potentially increase the risk of developing such complications. PMID- 12635685 TI - Herpes simplex virus type 2 acute retinal necrosis in an immunocompetent patient. AB - PURPOSE: To report a case of acute retinal necrosis caused by herpes simplex virus 2 in an otherwise healthy patient. CASE REPORT: A 45-year-old man presented with one month's history of decreased vision in the right eye. He had previously received a course of intravenous gancyclovir because of a clinical suspicion of cytomegalovirus retinitis. The patient's ocular history was remarkable for a similar episode in the left eye thirty years earlier, resulting in important visual impairment. System and laboratory investigations were unremarkable. Ocular examination showed severe anterior granulomatous uveitis, vitreous haze, areas of necrosis and retinal exudates. The anterior chamber tap disclosed the presence of HSV type 2, and oral steroids and acyclovir were instituted. Two weeks after the patient had been discharged, a retinal detachment occurred in the right eye, necessitating surgical repair. The presence of HSV type 2 was confirmed in the vitreous. Visual acuity recovered completely after surgery and the patient was placed on a maintenance dose of oral acyclovir. CONCLUSIONS: HSV type 2 is a rare cause of acute retinal necrosis in healthy patients. Bilateral involvement can occur in the fellow eye, even with a long delay. Acute retinal necrosis is a severe ocular inflammatory syndrome associated with a very poor visual outcome. It is caused by VZV, HSV type 1 and, less commonly, by HSV type 2. The disease can affect healthy patients and cause bilateral involvement in the fellow eye, even with a long delay. PMID- 12635686 TI - Perfluorohexyloctane endotamponade for treatment of persisting macular hole. AB - PURPOSE: To report the use of perfluorohexyloctane to endotamponade a persisting macular hole. METHODS: A 62-year-old patient presented with persisting stage IV macular hole, after uncomplicated, but unsuccessful, pars plana vitrectomy with peeling ok the inner limiting membrane of the retina for treatment of the disorder. A second pars plana vitrectomy was performed in combination with cataract surgery, using perfluorohexyloctane as a new heavy-weight ocular endotamponading agent. No additional membrane peeling was done. RESULTS: Four weeks later, marked emulsification of the perfluorohexyloctane and pronounced opacification of the posterior lens capsule were observed. After removal of the perfluorohexyloctane ten weeks after instillation, the macular hole was closed, and visual acuity increased from 0.1 to 0.3. The whole retinal surface was covered with a whitish amorphous membranous layer, that could partially be sucked off. After another four weeks, the membrane had mostly disappeared, and visual acuity increased to 0.5. CONCLUSIONS: Perfluorohexyloctane merits further evaluation for ocular endotamponade in patients with persisting macular holes. Formation of epiretinal membranes needs to be thoroughly investigated. PMID- 12635687 TI - Spontaneous visual improvement in pituitary metastasis. AB - PURPOSE: To report spontaneous visual improvement in a patient with unilateral optic neuropathy due to pituitary metastasis. METHODS: Report of a case. RESULTS: A 54-year-old woman with a history of breast carcinoma lost vision in her right eye to 20/70 without any other symptoms. Six days later, vision spontaneously improved to 20/30. A pituitary mass compressing the right intracranial optic nerve was found on magnetic resonance imaging and the diagnosis of metastatic breast carcinoma was confirmed by biopsy. CONCLUSIONS: Spontaneous visual improvement can occur in the setting of compressive optic neuropathy by a solid mass. PMID- 12635689 TI - Exhale while lifting or straining to avoid Valsalva retinopathy or bleeding from stressed retinal vessels. PMID- 12635688 TI - Liposarcoma metastatic to the orbit. AB - PURPOSE: To describe a patient with liposarcoma metastatic to the left orbit. METHODS: A 72-year-old man was admitted with diplopia and proptosis of the left eye. Previously, a retroperitoneal liposarcoma had been surgically removed followed by postoperative radiation. Visual acuity was normal. There was proptosis of the left eye, increased retrobulbar resistance and reduced mobility. Trans-septal orbital biopsies showed liposarcoma. The patient was initially treated with prednisolone and later received radio- and chemotherapy. Despite treatment he lost vision of the left eye and died a few months later. RESULTS: Histological examination of the orbital tumor revealed a spindle-cell liposarcoma similar to the primary tumor of the retroperitoneum. CONCLUSIONS: Liposarcoma metastatic to the orbit is rare, but should be suspected in a patient with proptosis caused by a space-occupying lesion and a history of liposarcoma. PMID- 12635690 TI - Chemopreventive effect of orange oil on the development of hepatic preneoplastic lesions induced by N-nitrosodiethylamine in rats: an ultrastructural study. AB - Orange peel oil is used extensively as an approved flavour enhancer in fruit drinks, carbonated beverages and as a scenting agent in soaps and cosmetics. Limonene, which is a monocyclic monoterpene is present in orange peel oil from 90 to 95% (w/w). Monoterpenes have been shown to be very effective chemopreventive agents against several rodent tumors and are currently in clinical trials. However, not much information is available regarding the ultrastructural changes associated with the chemopreventive effects of the monoterpenes. The effect of orange oil on the suppression of preneoplastic hepatic lesions during N nitrosodiethylamine (DEN) induced hepatocarcinogenesis was studied electron microscopically. Rats were administered 200 ppm DEN through drinking water for a period of 1 month. After an interval of 2 weeks, the animals were administered orange oil by gavage for a period of 5 1/2 months. The chemopreventive effect of orange oil was monitored on the basis of liver weight profile, histological pattern by light microscopy and ultrastructural alterations by electronmicroscopy. Orange oil administration following DEN treatment showed decreased liver weights, increased intercellular gap junctional complexes, cell density and polarity when compared with only the DEN treated rats. In the present study chemopreventive effect of orange oil on DEN-induced hepatic preneoplasia in rats which is associated with the restoration of the normal phenotype and upregulation of junctional complexes has been demonstrated. PMID- 12635691 TI - Photodynamic action of merocyanine 540 on carcinoma of cervix cells. AB - Results of the studies carried out on localization and photodynamic action of merocyanine 540 (MC540) on carcinoma of cervix (HeLa) cells are presented. Fluorescence microscopic study showed that when HeLa cells were incubated with MC540 in dark, the dye localized in plasma membrane of cells. Photoirradiation of cells in presence of MC540 led to enhancement of dye uptake, intracellular localization of dye and a dose dependent decrease in cell survival. Clonogenic assay showed 96% cell killing at a light dose of 42 kJ/m2. Photosensitization of cells resulted in loss of membrane integrity, decrease in plasma membrane fluidity and reduction in mitochondrial dehydrogenase activity as measured by tetrazolium reduction (MTT) assay. At a given light dose, the relative change in plasma membrane properties was higher than the reduction in activity of mitochondrial enzyme. These results suggest plasma membrane is a primary target of photosensitization of HeLa cells by MC540. PMID- 12635693 TI - Immunotoxicity of soluble and insoluble salts of cadmium instilled intratracheally. AB - Rats were intratracheally (i.t.) exposed to 36.5 or 27.5 microg of cadmium (Cd) as soluble cadmium chloride (CdCl2) and insoluble cadmium oxide (CdO) salts. The retention of metal in lungs, liver and kidney was assessed by atomic adsorption spectrophotometer. The animals were intraperitoneally (i.p.) primed with sheep red blood cells (SRBC) and assessed for the number of antibody forming cells in lung associated lymph nodes (LALN) and spleen. Both the compounds had similar retention of metal in lungs but CdO induced more pulmonary inflammatory and degradative changes than CdCl2. The larger influx of polymorphonuclear cells (PMNs) following CdO exposure appears to be due to the absence of protection afforded by Cd induced metallothionein cytoplasmic protein while the Cd metallothionein complex formed in the case of CdCl2 is more protective. However both forms of Cd had similar local immunosuppressive potential but CdO had more prolonged suppressive effect. PMID- 12635692 TI - In vivo inhibition of selenium dependent glutathione peroxidase and superoxide dismutase in rats by diethyldithiocarbamate. AB - Intraperitoneal injection of rats with diethyldithiocarbamate (1.2 g/kg body wt) led to maximum diminution of superoxide dismutase activity at 1 hr by 86 and 84% in liver and red blood cell respectively with a gradual return to the normal level at 48 hr after administration of injection. Significant inhibition of selenium-dependent glutathione peroxidase was also observed, which returned to normal at 48 hr after administration of injection. However, maximum decline in its activity was at 12 hr by 52 and 73% in liver and red blood cells respectively. No significant difference in tissue level of selenium-independent glutathione peroxidase was observed during time course study after diethyldithiocarbamate administration. It is possible that inhibition of superoxide dismutase by diethyldithiocarbamate leads to accumulation of superoxide anion which in turn inactivates selenium-dependent glutathione peroxidase by its reaction with selenium at the active site of the enzyme. PMID- 12635694 TI - Effect of fish oil on mitochondrial respiration in isoproterenol induced myocardial infarction in rats. AB - Following isoproterenol treatment mitochondrial lipid peroxidation, phosphoslipase activity, lactate and calcium increased significantly, while activities of tricarboxylic acid cycle enzymes, enzymes of respiratory chain and ATP production showed decline. Oxidative phosphorylation was also affected on isoproterenol treatment with significant reduction in all the variables. Fish oil pretreatment in isoproterenol treated rats showed improved mitochondrial energy metabolism. The results suggest cardioprotective effect of fish oil. PMID- 12635695 TI - Amelioration of experimental diabetic neuropathy and gastropathy in rats following oral administration of plant (Eugenia jambolana, Mucuna pruriens and Tinospora cordifolia) extracts. AB - Extract of M. charantia (200 mg/kg), E. jambolana (200 mg/kg), M. pruriens (200 mg/kg) and T. cordifolia (400 mg/kg) was administered for 50 days in STZ induced diabetic mice, the plasma glucose concentration was reduced by 24.4, 20.84, 7.45 and 9.07% respectively. Tail flick latency (TFL) and gastric transit percentage were significantly higher in diabetic controls versus normal controls. M. charantia and E. jambolana modified it favorably while M. pruriens and T. cordifolia did not exert any favorable change. PMID- 12635696 TI - Effect of co-administration of piperine on pharmacokinetics of beta-lactam antibiotics in rats. AB - Co-administration of piperine, an alkaloid isolated from Piper nigrum L. enhanced bioavailability of beta lactam antibiotics, amoxycillin trihydrate and cefotaxime sodium significantly in rats. The improved bioavailability is reflected in various pharmacokinetic parameters viz. tmax, Cmax, t(1/2) and AUC, of these antibiotics. The increased bioavailability could be attributed to the effect of piperine on microsomal metabolising enzymes or enzymes system. PMID- 12635697 TI - Cell proliferation and natural killer cell activity by polyherbal formulation, Immu-21 in mice. AB - Immunomodulatory activity of an Ayurvedic polyherbal formulation, Immu-21 containing extracts of Ocimum sanctum, Withania somnifera, Emblica officinalis and Tinospora cordifolia was studied on proliferative response of splenic leukocytes to T cell mitogens, concanavalin (Con)-A and phytohemagglutinin (PHA) and B cell mitogen, lipopolysaccharide (LPS) in vitro by [3H]-thymidine uptake assay in mice. The cytotoxic activity of Immu-21 was tested by measuring the splenic leukocyte natural killer (NK) cell activity against K 562 cells. Intraperitoneal (i.p.) treatment with Immu-21 (30 mg/kg) once a day for 14 and 21 days did not cause change in body weight and spleen weight, where as splenocytes/spleen count was increased. Treatment of Immu-21 (30 mg/kg, i.p.) for 14 days and 1 mg/kg for 21 days significantly increased LPS induced leukocyte proliferation. NK cell activity was significantly increased when mice were pretreated with Immu-21 (10 and 30 mg/kg, i.p.) once a day for 7 days. The results indicate that pretreatment with Immu-21 selectively increased the proliferation of splenic leukocyte to B cell mitogen, LPS and cytotoxic activity against K 562 cells in mice. PMID- 12635698 TI - Immunological identification of two female-specific proteins from the plasma of Indian freshwater murrel, Channa punctatus (Bloch). AB - Existence of a non-phosphorylated female-specific protein (FS II), in addition to phosphorylated vitellogenin (FS I), in the plasma of murrel by exogenous administration of estradiol-17beta is reported. Polyspecific rabbit antibodies were raised against estrogen-inducible murrel plasma proteins. This antiserum was absorbed with normal male serum in order to obtain female-specific antiserum (FSAS). Radial immunodiffusion studies suggested that both the proteins (FS I and FS II) were present in the plasma of E2-treated and normal vitellogenic females and in the ovarian homogenate from gravid females, but absent in normal male plasma. Autoradiographic experiments demonstrated that phosphorus moiety was attached with FS I only. Further, immunoelectrophoretic analysis and peptide maps supported the observation that FS I and FS II were discrete, unrelated female specific proteins. PMID- 12635699 TI - Evaluation of guinea pig model for experimental Salmonella serovar Abortusequi infection in reference to infertility. AB - The present study conclusively revealed the role for Salmonella enterica subspecies enterica serovar Abortusequi in conception failure. None of the 12 guinea pigs conceived when orally exposed to sublethal dose of the pathogen during breeding, while 66.67% of animals in control group were found pregnant during same period of observation under similar conditions. Salmonella carrier animals also had drastic reduction in conception rate (16.67%). During mid pregnancy, S. Abortusequi exposure to guinea pigs through intravaginal, intramuscular and subcutaneous routes induced fetal death followed by resorption. While 2 out of 6 orally inoculated and 3 out of 6 intraperitonially inoculated guinea pigs aborted, in rest of the animals fetal death was followed by meceration and resorption. It was interesting to note that S. Abortusequi could not persist longer than a week in males while in pregnant females it could be detected for >10 weeks after inoculation. In late pregnancy, most of the exposed animals aborted and non aborting animals though had normal parturition, survival rate of their babies was nearly zero in comparison to the control group. The study revealed role for S. Abortusequi in impairing conception, abortion, early fetal deaths, fetal meceration and resorption. Further studies are required to identify factors responsible for increased susceptibility of females particularly during pregnancy. PMID- 12635700 TI - Artocarpus heterophyllus seeds inhibits sexual competence but not fertility of male rats. AB - According to Ayurvedic literature of Sri Lanka, roasted seeds of Artocarpus heterophyllus Lam. (Family: Moraceae) has aphrodisiac activity. However, some reproductively active young men in rural areas of Sri Lanka claim that consumption of these seeds few hours prior to coitus disrupts sexual function. Because of these two conflicting claims, it was thought useful to scientifically investigate the effects of A. heterophyllus seeds on male sexual function and fertility. This was done using a seed suspension in 1% methylcellulose (SS) in rats. In a sexual behaviour study using receptive female rats, an oral administration of 500 mg/kg dose of SS markedly inhibited libido, sexual arousal, sexual vigour and sexual performance within 2 hr. Further, the treatment induced a mild erectile dysfunction. These antimasculine effects on sexual function was not evident 6 hr post treatment indicating rapid onset and offset of action. Further, these actions on the sexual behaviour was not due to general toxicity, liver toxicity, stress or reduction in blood testosterone level but due to marked sedative activity. In a mating study, SS failed to alter ejaculating competence and fertility. These results suggest that A. heterophyllous seeds do not have aphrodisiac action, at least, in rats. PMID- 12635701 TI - Marker assisted detection of gene (1Dx5) and translocation (1B/1R) in wheat genotypes. AB - Detection of 1Dx5 gene and presence of 1B/1R wheat rye translocation were studied in nineteen elite Indian wheat genotypes using AS-PCR and STS markers, respectively. Fifteen genotypes had 1B/1R translocation whereas ten showed presence of 1Dx5 gene. More than 50 per cent of the genotypes tested were found positive for both 1Dx5 and 1B/1R translocation. The results are in conformity with HMW glutenin SDS-PAGE profile for 1Dx5 and cytological observations for 1B/1R translocation. PMID- 12635702 TI - Fungistatic activity of Semecarpus anacardium Linn. f nut extract. AB - Alcoholic extract of dry nuts of S. anacardium showed dose dependent antifungal activity in vitro against Aspergillus fumigatus and Candida albicans. At 400 mg/ml concentration, growth of both the fungi was inhibited and considerable reduction in size of cells and hyphae was observed. Sporulation also decreased. PMID- 12635703 TI - Bioleaching of copper from chalcopyrite ore by fungi. AB - Microorganisms have been geologically active in mineral formation, mineral diagenesis and sedimentation via direct action of their enzymes or indirectly through chemical action of their metabolic products. This property of microorganisms is being harnessed during the recent years for extraction of metals from their ores, especially from low-grade ores. In the present study bioleaching of copper from its low-grade chalcopyrite ore using 26 isolates of acidophilic fungi is reported. Most of these fungal strains belonged to the genera Aspergillus, Penicillium and Rhizopus. The leaching experiments were conducted in Czepek Dox minimal medium containing 1% (100 mesh) ore with shaking at room temperature for 20 days. Out of these, 4 isolates exhibited significant bioleaching activities. Maximum leaching of copper (78 mg/L) was observed with Aspergillus flavus (DSF-8) and Aspergillus niger (DOF-1). Nutritional and environmental conditions for optimum bioleaching were standardized. Present study indicates the usefulness of acidophilic fungi in bioleaching of copper from its low-grade ores. PMID- 12635704 TI - Fermentation of starch to ethanol by an amylolytic yeast Saccharomyces diastaticus SM-10. AB - A total of fifteen yeast strains were isolated from natural sources including fruits, soil, molasses, honey and a variety of indigeneous fermented foods. Screening of these strains for growth, ethanol production and glucoamylase activity led to selection of a yeast strain SM-10 identified as S. diastaticus having maximum glucoamylase activity (80 units ml(-1)) and ethanol production from starch (3.5%). Ethanol production from wheat flour was found to be 1.75% which could be increased to 5.2% after treatment of wheat flour with pepsin, diastase and glucoamylase. PMID- 12635705 TI - In vitro organogenesis and genetic transformation in popular Cucumis sativus L. through Agrobacterium tumefaciens. AB - The effect of growth regulators and culture conditions on the morphogenetic response of cotyledonary leaf discs was studied in popular cucumber variety (Cucumis sativus cv. Sheetal). Organogenesis was induced directly without any intervening callus phase on Murashige and Skoog medium supplemented with different concentrations of benzyladenine and indole propionic acid. Best results (93%) were obtained in the presence of the 4 mg/L benzyladenine and 1 mg/L IPA. The elongated shoots were rooted in basal medium with 1 mg/L indole butyric acid, hardened and transferred to the field conditions. Genetic transformation system has been established for Cucumis sativus cv. Sheetal, plants by infecting cotyledonary explants with Agrobacterium tumefaciens strain LBA4404 carrying binary plasmid pBI121, which contains scorable marker, beta-glucuronidase and selectable marker nptII under the CaMV 35S promoter. Infection was most effective when explants were infected with Agrobacterium for 15 min and co-cultivated for 2 days in the co-cultivation medium. Shoots were regenerated directly from cotyledonary leaf explants in the presence of kanamycin (50 microg/ml) and analysed. Southern blot analysis confirmed that transformation had occurred. This method will allow genetic improvement of this crop by the introduction of agronomically important genes. PMID- 12635706 TI - Host plant nodule parameters associated with nitrogen fixation efficiency in French bean (Phaseolus vulgaris L) cultivars. AB - Two cultivars of French bean (Phaseolus vulgaris L.) viz. contender and arka komal were planted in polythene bags containing sand and grown under glasshouse conditions. The nodulation status, shoot/root biomass, activities of several nodule enzymes, total soluble protein and leghaemoglobin contents were monitored over the entire growth period. Allantoinase activity in leaves was measured to monitor the ureide degrading capacity. Significant genotype difference was observed in both the cultivars. All the parameters showed a decline after flowering except uricase, which declined before flowering. Malate dehydrogenase and isocitrate dehydrogenase showed a constant decline throughout the growth period. Degree of decline varied with the genotype for all the parameters. Leghaemoglobin content, PEP carboxylase activity and ureide degrading capacity of leaves did not show an appreciable decline in contender and were significantly higher than in arka komal. These factors can be used to increase nitrogen fixation in French bean. PMID- 12635707 TI - Anti-angiogenic activity of BIM (bio-immunomodulator). AB - Angiogenesis and immune suppression are the two important factors responsible for embryo implantation and development of tumor. Therefore, disruption of angiogenesis and upliftment of immune function is essential for control of tumor growth as well as to regulate the activity of post coital contraceptives. BIM is an immunomodulatory cytokine obtained from rodent bone marrow that showed anti implantation and anti-tumor activities. It also improved T/B cell and monocyte macrophage functions. In this communication the anti-angiogenic property of BIM is demonstrated in pregnant rat model. BIM induced disintegration of uterine myometrium and blood vessels. No implantation was observed compared to control. It is proposed that depending upon the physiological condition of the host BIM could modulate host immune function and exert its anti-angiogenic effect. PMID- 12635708 TI - Effect of enrofloxacin on AgNOR counts in chick bone marrow nuclei. AB - The mAgNOR and pAgNOR counts reflecting the number of rDNA genes being transcribed, showed a highly significant increase from control values following administration of 5X, 10X, 15X doses of enrofloxacin in chicken. The maximum increase for both, mAgNOR and pAgNOR was shown by birds receiving 15X dose, sacrificed 48 hr after the last drug injection. The increase of pAgNOR at 5X (24 hr) was not significant relative to control values. After 72 hr time interval, AgNOR counts were not feasible due to poor differentiation. Values at 15X (24 hr) showed a decrease in number of AgNOR counts (non-significant) relative to 10X, probably due to depression of transcriptional activity of rDNA genes, which, however, is removed at 48 hr. The general increase in mAgNOR and pAgNOR with dose reflects hypertranscription of rDNA genes so that the birds can cope up with the drug induced toxicity. PMID- 12635709 TI - Effect of oral magnesium supplementation on experimental pre-eclampsia induced by prolonged blockade of nitric oxide synthesis in pregnant rats. AB - Nitric oxide inhibitor L-NAME when given alone caused a significant rise in both systolic and diastolic pressure, an increase in 24 hr urinary protein excretion and reduction in weight of the litter as compared to control group. Supplementation of MgSO4 at lower dose (250 mg/kg) did not inhibit this pre eclamptic effect of L-NAME; but in higher doses (500 and 750 mg/kg), it inhibited the pre-eclamptic action of L-NAME. The results suggest that administration of MgSO4 improves the foetal outcome and significantly prevents the development of symptoms of pre-eclampsia like hypertension and proteinuria. PMID- 12635710 TI - Measurement of creatinine by Jaffe's reaction--determination of concentration of sodium hydroxide required for maximum color development in standard, urine and protein free filtrate of serum. AB - Creatinine in serum or urine is determined by Jaffe's reaction where creatinine produces quantitatively an orange color with picric acid in alkaline medium. After allowing an incubation time of 15 min at room temperature for color development the color is measured at 520 nm. Without taking into consideration the acidic nature of standard, protein free filtrate (PFF) of serum and urine, 1% picric acid and 0.75N NaOH are used in this reaction for color development in standard, PFF of serum and urine. An investigation was thought to be necessary to determine the optimum alkali concentration required in standard, PFF of serum and urine. The results show that 0.25, 0.75 and 1N NaOH give maximum color in urine, standard and PFF of serum respectively. A standard solution of creatinine is prepared in 0.1N HCl and the PFF of serum is obtained by addition of fresh tungstic acid. Alkali is consumed to neutralise the acids in both these cases. For urine creatinine measurement, a direct diluted urine sample is used. The difference in the requirement of NaOH is conceivable. The routine use of 0.75N NaOH irrespective of the nature of specimen as is done in all biochemical laboratories, for creatinine measurement needs modification in the light of this investigation. PMID- 12635711 TI - Effect of dietary protein on vitamin A levels in plasma and liver of hypervitaminotic-A rats. AB - Level of vitamin A increased in plasma and liver in hypervitaminotic A albino rats fed normal quantity of protein in diet. In low protein fed state vitamin A level in liver increased due to accumulation of vitamin A and lack of carrier protein with an associated decrease of plasma vitamin A. In high protein fed rats the level of vitamin A in plasma increased due to enhanced transport while in liver it decreased. The results indicate that for normal transport of vitamin A adequate plasma protein level is essential. PMID- 12635712 TI - Male accessory gland secretions in hybrids of Drosophila nasuta nasuta and D. n. albomicans neither show luxuriance nor breakdown. AB - Male accessory gland secretions, which have a role to play in reproduction have been investigated. The number of cells that make up the gland, the quantity of secretions synthesized and the influence of these secretions on fecundity of the female have been studied in D. n. nasuta, D. n. albomicans and their F1 progeny. The results revealed that the hybrid males show a trend towards D. n. nasuta in the synthesis of male accessory gland proteins and the fecundity of the female is influenced more by its genetic constitution rather than the quantity of accessory gland secretions. PMID- 12635713 TI - Evaluation of mosquitocidal activity of Annona squamosa leaves against filarial vector mosquito, Culex quinquefasciatus Say. AB - Methanolic extract of leaves of A. squamosa was tested for mosquitocidal effect against C. quinquefasciatus. A liquid mosquito insecticide formulation was prepared with the extract (1, 3 and 5 %w/w) using deodorized kerosene as solvent and investigated for its knock-down and 24 hr mortality. The extract formulation produced dose dependent activity, exhibited significantly shorter knock down KD50 and KD90 values and produced significant mortality. The results suggest the potential mosquitocidal effect of A. squamosa on C. quinquefasciatus. PMID- 12635714 TI - Effect of a non-hormonal feed additive, Cholymbi on growth, body composition and digestive enzyme activity of common carp, Cyprinus carpio. AB - Supplementation of pelleted feeds (30% crude protein) consisting of practical ingredients with 250, 375 and 500 mg Cholymbi/kg diet resulted significantly improved the growth of common carp, Cyprinus carpio. Highest final average weight was recorded with diet containing 500 mg Cholymbi/kg. Feeding Cholymbi to fish showed increased carcass protein and fat. Gut protease and hepatopancreas amylase activity were the highest with 500 mg Cholymbi/kg incorporated diet (T3). The results of the present study suggest that Cholymbi, a non-hormonal growth promoter could be effectively used in the diet of common carp. Further investigations are required to work out the optimal dosage required in the diet of common carp. PMID- 12635715 TI - Influence of cell wall degrading enzymes on colonization of N2 fixing bacterium, Azorhizobium caulinodans in rice. AB - In rice, nodule like structures were formed by inoculation of A. caulinodans combined with growth regulators and enzymes. Among the treatments, combination of cell wall degrading enzyme mixture and NAA with A. caulinodans induced more number of paranodules in rice. Total nitrogen content also increased in treated plants compared to uninoculated control. PMID- 12635716 TI - Early diagnosis of Alzheimer's disease. PMID- 12635717 TI - Towards therapeutic angiogenesis. PMID- 12635718 TI - The precautionary principle versus risk analysis. AB - Precautionary behavior serves a valuable protective purpose as a first response to the new. It causes a quick retreat to the safety of the familiar; it provides time for a realistic "friend or foe" assessment of a new event. This rational assessment is often delayed by the sluggishness of government bureaucratic processes, or stopped by an implied challenge to a status quo. At the public level, reassurance may be slow to overcome an early uncertainty. However, a precautionary response does not provide an operational governing principle, although it makes publicly plausible an indefinite concealment of de facto political actions, or nonaction. The alternative of rational decision making at the policy level should flow from a comparative benefit/cost/risk analysis. Such early risk analyses have pragmatic uncertainties based on the limited available knowledge base and, accordingly, require judgmental application. PMID- 12635719 TI - A revised economic analysis of restrictions on the use of cell phones while driving. AB - Evidence that cell phone use while driving increases the risk of being involved in a motor vehicle crash has led policymakers to consider prohibitions on this practice. However, while restrictions would reduce property loss, injuries, and fatalities, consumers would lose the convenience of using these devices while driving. Quantifying the risks and benefits associated with cell phone use while driving is complicated by substantial uncertainty in the estimates of several important inputs, including the extent to which cell phone use increases a driver's risk of being involved in a crash, the amount of time drivers spend using cell phones (and hence their aggregate contribution to crashes, injuries, and fatalities), and the incremental value to users of being able to make calls while driving. Two prominent studies that have investigated cell phone use while driving have concluded that the practice should not be banned. One finds that the benefits of calls made while driving substantially exceed their costs while the other finds that other interventions could reduce motor vehicle injuries and fatalities (measured in terms of quality adjusted life years) at a lower cost. Another issue is that cell phone use imposes increased (involuntary) risks on other roadway users. This article revises the assumptions used in the two previous analyses to make them consistent and updates them using recent data. The result is a best estimate of zero for the net benefit of cell phone use while driving, a finding that differs substantially from the previous study. Our revised cost-effectiveness estimate for cell phone use while driving moves in the other direction, finding that the cost per quality adjusted life year increases modestly compared to the previous estimate. Both estimates are very uncertain. PMID- 12635721 TI - Joint persistence of transformation products in chemicals assessment: case studies and uncertainty analysis. AB - The joint persistence (JP) quantifies the environmental persistence of a parent compound and a selection of relevant transformation products. Here, the importance as well as the uncertainty of the JP in comparison to the persistence of the parent compound alone (primary persistence, PP) are investigated. To demonstrate the effect of transformation products on the environmental persistence of organic chemicals, three case studies of parent compounds (nonylphenol ethoxylates, perchloroethylene, atrazine) and transformation products are investigated in detail with a multimedia fate model. Comparison of the PP and JP values shows that transformation products can significantly increase the persistence. In addition to the point estimates of PP and JP, the associated uncertainties are investigated. For each of the case studies, the chemical-specific input parameters of all compounds are varied and the corresponding variance of the PP and JP is determined by Monte Carlo simulations. Interestingly, the higher number of input parameters required for the JP does not necessarily increase the uncertainty of the JP as compared to that of the PP alone. An exact mathematical expression specifying the contribution of each transformation product to the JP is given. When transformation products are grouped in different generations, it becomes discernible that the first generation increases the JP most; the later generations are of decreasing importance. Finally, the effect of incomplete knowledge of the transformation products and their properties on the JP results is discussed. For reliable JP estimates, knowledge of the first generation transformation products and their degradation rate constants is required. PMID- 12635720 TI - Assessing cancer risks from short-term exposures in children. AB - For the vast majority of chemicals that have cancer potency estimates on IRIS, the underlying database is deficient with respect to early-life exposures. This data gap has prevented derivation of cancer potency factors that are relevant to this time period, and so assessments may not fully address children's risks. This article provides a review of juvenile animal bioassay data in comparison to adult animal data for a broad array of carcinogens. This comparison indicates that short-term exposures in early life are likely to yield a greater tumor response than short-term exposures in adults, but similar tumor response when compared to long-term exposures in adults. This evidence is brought into a risk assessment context by proposing an approach that: (1) does not prorate children's exposures over the entire life span or mix them with exposures that occur at other ages; (2) applies the cancer slope factor from adult animal or human epidemiology studies to the children's exposure dose to calculate the cancer risk associated with the early-life period; and (3) adds the cancer risk for young children to that for older children/adults to yield a total lifetime cancer risk. The proposed approach allows for the unique exposure and pharmacokinetic factors associated with young children to be fully weighted in the cancer risk assessment. It is very similar to the approach currently used by U.S. EPA for vinyl chloride. The current analysis finds that the database of early life and adult cancer bioassays supports extension of this approach from vinyl chloride to other carcinogens of diverse mode of action. This approach should be enhanced by early-life data specific to the particular carcinogen under analysis whenever possible. PMID- 12635722 TI - Canada Wide Standard for particulate matter and ozone: cost-benefit analysis using a Life Quality Index. AB - The adverse impacts of particulate air pollution and ground-level ozone on public health and the environment have motivated the development of Canada Wide Standards (CWS) on air quality. In cost-benefit analysis of air-quality options, valuation of reduction in mortality is a critical step as it accounts for almost 80% of the total benefits and any bias in its evaluation can significantly skew the outcome of the analysis. The overestimation of benefits is a source of concern since it has the potential of diverting valuable resources from other needs to support broader health care objectives, education, and social services that contribute to enhanced quality of life. We have developed a framework of reasoning for the assessment of risk-reduction initiatives that would support the public interest and enhance safety and quality of life. This article presents the Life Quality Index (LQI) as a tool to quantify the level of expenditure beyond which it is no longer justifiable to spend resources in the name of safety. It is shown that the LQI is a compound social indicator comprising societal wealth and longevity, and it is also equivalent to a utility function consistent with the basic principles of welfare economics and decision analysis. The LQI approach overcomes several shortcomings of the method used by the CWS Development Committee and provides guidance on the compliance costs that can be justified to meet the Standards. PMID- 12635723 TI - PER pressure: New Jersey's "Population Emissions Ratio" environmental equity screening model. AB - In recent years, much attention has focused on how to incorporate environmental equity considerations into government permitting programs for environmentally regulated facilities. On February 4, 2002, the New Jersey Department of Environmental Protection (NJDEP) became the first state environmental agency to propose a broad legally binding rule intended to guard against environmental inequity in its permitting decisions. The proposed rule centered on an innovative computerized environmental equity (EE) screening model that used "Population Emissions Ratios" to identify small geographic areas in which environmental equity concerns might exist and to simulate the effect on statewide environmental equity of increasing environmental risks in small geographic areas. The NJDEP's model examined an extensive array of ethnic groups, included a variety of environmental risks, evaluated most of those risks in terms of human health, and used an innovative simulation process designed to identify permitting decisions that would worsen statewide environmental inequity. The results of the NJDEP's efforts, however, pose substantial concerns. For example, some key provisions of the NJDEP's model were inadequately explained and some were illogical and would bias its results. The model might be susceptible to generating implausible results due to small, meaningless, and/or essentially random fluctuations in its data inputs. The model used relatively large geographic areas as the units of analysis and interpolated results between them, rather than using smaller geographic areas and avoiding interpolation errors. Finally, the environmental risks evaluated by the model were both arguably over- and underinclusive. Thus, the NJDEP's efforts, although noteworthy, raised more issues than they settled. PMID- 12635724 TI - Incremental and average cost-effectiveness ratios: will physicians make a distinction? AB - Physicians are increasingly asked to use cost-effectiveness information when evaluating alternative health care interventions. Little is known about how the way such information is presented can influence medical decision making. We presented physicians with hypothetical screening scenarios with multiple options, varying the type of cost-effectiveness ratios provided as well as whether the scenarios described cancer screening settings that were familiar or unfamiliar. Half the scenarios used average cost-effectiveness ratios, as commonly reported, calculating benefits and costs relative to a no-screening option. The other half used the preferred incremental cost-effectiveness ratios, with each option's benefits and costs calculated relative to the next best alternative. Relative to average cost-effectiveness ratios, incremental cost-effectiveness information significantly reduced preference for the most expensive screening strategies in two of three unfamiliar scenarios. No such difference was found for familiar scenarios, for which physicians likely have established practice patterns. These results suggest that, in unfamiliar settings, average cost-effectiveness ratios as reported in many analyses reported in the literature can hide the often high price for achieving incremental health care goals, potentially causing physicians to choose interventions with poor cost effectiveness. PMID- 12635725 TI - Communicating air quality information: experimental evaluation of alternative formats. AB - A long-running effort in environmental communication is daily publication of a report on local air pollution in many American newspapers based on the Pollutant Standards Index (PSI). A 1998 proposal by the U.S. Environmental Protection Agency (U.S. EPA) to change the PSI prompted a survey experiment with 1,100 adults in Philadelphia, evaluating the proposed change's ability to better inform the populace. The effects of exposure to the old and new versions of the PSI, as well as health cautions and information about groups sensitive to air pollution, were compared with evaluation criteria suggested by Weinstein and Sandman (1993). Sample respondents had strong baseline concerns about air pollution. Descriptors of air quality (e.g., "good; " "unhealthy") were difficult to discriminate, particularly in the New format. Concern rose as hypothetical air pollution levels rose, but the New format (as well as PSI versions without health cautions or sensitive-group information) evoked a sharp discontinuity in concern between below- and above-standard pollution levels. Both Old and New formats reduced concern relative to no provision of PSI information at all, but the New format reduced concern significantly more than the Old version. No PSI format did particularly well at increasing knowledge of air pollution or decreasing intentions to be active outdoors during high pollution, contrary to the agency's aim. Although U.S. EPA has since adopted the new proposal as a national "Air Quality Index" requirement, the experiment's results illuminate the strengths and limitations of the new PSI as a means of informing citizens and motivating them to protect themselves. PMID- 12635727 TI - Methods and rationale for derivation of a reference dose for methylmercury by the U.S. EPA. AB - In 2001, the U.S. Environmental Protection Agency derived a reference dose (RfD) for methylmercury, which is a daily intake that is likely to be without appreciable risk of deleterious effects during a lifetime. This derivation used a series of benchmark dose (BMD) analyses provided by a National Research Council (NRC) panel convened to assess the health effects of methylmercury. Analyses were performed for a number of endpoints from three large longitudinal cohort studies of the neuropsychological consequences of in utero exposure to methylmercury: the Faroe Islands, Seychelles Islands, and New Zealand studies. Adverse effects were identified in the Faroe Islands and New Zealand studies, but not in the Seychelles Islands. The NRC also performed an integrative analysis of all three studies. The EPA applied a total uncertainty factor (UF) of 10 for intrahuman toxicokinetic and toxicodynamic variability and uncertainty. Dose conversion from cord blood mercury concentrations to maternal methylmercury intake was performed using a one-compartment model. Derivation of potential RfDs from a number of endpoints from the Faroe Islands study converged on 0.1 microg/kg/day, as did the integrative analysis of all three studies. EPA identified several areas for which further information or analyses is needed. Perhaps the most immediately relevant is the ratio of cord:maternal blood mercury concentration, as well as the variability around this ratio. EPA assumed in its dose conversion that the ratio was 1.0; however, available data suggest it is perhaps 1.5-2.0. Verification of a deviation from unity presumably would be translated directly into comparable reduction in the RfD. Other areas that EPA identified as significant areas requiring further attention are cardiovascular consequences of methylmercury exposure and delayed neurotoxicity during aging as a result of previous developmental or adult exposure. PMID- 12635728 TI - Differences in pharmacokinetics between children and adults--II. Children's variability in drug elimination half-lives and in some parameters needed for physiologically-based pharmacokinetic modeling. AB - In earlier work we assembled a database of classical pharmacokinetic parameters (e.g., elimination half-lives; volumes of distribution) in children and adults. These data were then analyzed to define mean differences between adults and children of various age groups. In this article, we first analyze the variability in half-life observations where individual data exist. The major findings are as follows. The age groups defined in the earlier analysis of arithmetic mean data (0-1 week premature; 0-1 week full term; 1 week to 2 months; 2-6 months; 6 months to 2 years; 2-12 years; and 12-18 years) are reasonable for depicting child/adult pharmacokinetic differences, but data for some of the earliest age groups are highly variable. The fraction of individual children's half-lives observed to exceed the adult mean half-life by more than the 3.2-fold uncertainty factor commonly attributed to interindividual pharmacokinetic variability is 27% (16/59) for the 0-1 week age group, and 19% (5/26) in the 1 week to 2 month age group, compared to 0/87 for all the other age groups combined between 2 months and 18 years. Children within specific age groups appear to differ from adults with respect to the amount of variability and the form of the distribution of half lives across the population. The data indicate departure from simple unimodal distributions, particularly in the 1 week to 2 month age group, suggesting that key developmental steps affecting drug removal tend to occur in that period. Finally, in preparation for age-dependent physiologically-based pharmacokinetic modeling, nationally representative NHANES III data are analyzed for distributions of body size and fat content. The data from about age 3 to age 10 reveal important departures from simple unimodal distributional forms-in the direction suggesting a subpopulation of children that are markedly heavier than those in the major mode. For risk assessment modeling, this means that analysts will need to consider "mixed" distributions (e.g., two or more normal or log normal modes) in which the proportions of children falling within the major versus highweight/fat modes in the mixture changes as a function of age. Biologically, the most natural interpretation of this is that these subpopulations represent children who have or have not yet received particular signals for change in growth pattern. These apparently distinct subpopulations would be expected to exhibit different disposition of xenobiotics, particularly those that are highly lipophilic and poorly metabolized. PMID- 12635729 TI - Challenges confronting risk analysis of potential thyroid toxicants. AB - Screening and testing for potential thyroid toxicants using endpoints of thyroid function, including circulating levels of thyroid hormones and thyrotropin, will not capture toxicants that directly interfere with thyroid hormone action at the receptor. The goals of the present review are to provide a critique of the literature focused on thyroid hormone and brain development as it relates to testing and evaluating thyroid toxicants, and to propose possible solutions to this perceived dilemma. PMID- 12635730 TI - Perspectives on incorporating human neurobehavioral end points in risk assessments. AB - Three topics are addressed: (1) measurement issues (e.g., the reliability and validity of neurobehavioral test scores), (2) general principles of assessment, including test selection, and (3) interpretation of scores. Psychological tests generally perform as well as medical tests in terms of reliability and validity. Test manuals, assessment textbooks, and psychologists are useful resources to the risk assessor. The variety of different tests employed in neurobehavioral studies complicates interstudy comparisons. In addition, tests that ostensibly assess the same general domain of function might assess somewhat different abilities within that domain. Although a uniform battery for use in all studies seems desirable, the battery appropriate for a specific study depends on study goals, knowledge about the mechanism(s) of neurotoxicity, nature of the study population, and pattern of exposure. Exposure-related neurobehavioral deficits are generally indicators of "altered function" rather than of "clinical disease." Limiting concern to end points corresponding to clinical disease might not be appropriate. Many neurobehavioral diagnoses are phenomenological and a neurotoxicant might cause a unique pattern of deficits for which no label has been created. The concern that a small shift in the central tendency of a distribution of test scores has no significance for the individual should be reexamined in light of the prevention paradox, formulated on the basis of epidemiologic studies of chronic disease. Poor performance on a neurobehavioral test does not necessarily map clearly onto underlying behavioral or neural substrate. The absence of such linkages, given current knowledge about brain-behavior relationships, should not reduce confidence in neurobehavioral end points. Use of neurobehavioral test scores involves considerations that differ little from those that the risk assessor routinely addresses in using end points commonly used in research on other topics in environmental epidemiology. PMID- 12635731 TI - Interactions of predictive microbiology and risk assessment. PMID- 12635732 TI - Modeling microbial growth within food safety risk assessments. AB - Risk estimates for food-borne infection will usually depend heavily on numbers of microorganisms present on the food at the time of consumption. As these data are seldom available directly, attention has turned to predictive microbiology as a means of inferring exposure at consumption. Codex guidelines recommend that microbiological risk assessment should explicitly consider the dynamics of microbiological growth, survival, and death in foods. This article describes predictive models and resources for modeling microbial growth in foods, and their utility and limitations in food safety risk assessment. We also aim to identify tools, data, and knowledge sources, and to provide an understanding of the microbial ecology of foods so that users can recognize model limits, avoid modeling unrealistic scenarios, and thus be able to appreciate the levels of confidence they can have in the outputs of predictive microbiology models. The microbial ecology of foods is complex. Developing reliable risk assessments involving microbial growth in foods will require the skills of both microbial ecologists and mathematical modelers. Simplifying assumptions will need to be made, but because of the potential for apparently small errors in growth rate to translate into very large errors in the estimate of risk, the validity of those assumptions should be carefully assessed. Quantitative estimates of absolute microbial risk within narrow confidence intervals do not yet appear to be possible. Nevertheless, the expression of microbial ecology knowledge in "predictive microbiology" models does allow decision support using the tools of risk assessment. PMID- 12635733 TI - Growth and heat resistance kinetic variation among various isolates of Salmonella and its application to risk assessment. AB - The abilities of cells of a particular type of bacteria to leave lag phase and begin the process of dividing or surviving heat treatment can depend on the serotypes or strains of the bacteria. This article reports an investigation of serotype-specific differences in growth and heat resistance kinetics of clinical and food isolates of Salmonella. Growth kinetics at 19 degrees C and 37 degrees C were examined in brain heart infusion broth and heat resistance kinetics for 60 degrees C were examined in beef gravy using a submerged coil heating apparatus. Estimates of the parameters of the growth curves suggests a small between serotype variance of the growth kinetics. However, for inactivation, the results suggest a significant between-serotype effect on the asymptotic D-values, with an estimated between-serotype CV of about 20%. In microbial risk assessment, predictive microbiology is used to estimate growth and inactivation of pathogens. Often the data used for estimating the growth or inactivation kinetics are based on measurements on a cocktail--a mixture of approximately equal proportions of several serotypes or strains of the pathogen being studied. The expected growth or inactivation rates derived from data using cocktails are biased, reflecting the characteristics of the fastest growing or most heat resistant serotype of the cocktail. In this article, an adjustment to decrease this possible bias in a risk assessment is offered. The article also presents discussion of the effect on estimating growth when stochastic assumptions are incorporated in the model. In particular, equations describing the variation of relative growth are derived, accounting for the stochastic variations of the division of cells. For small numbers of cells, the expected value of the relative growth is not an appropriate "representative" value for actual relative growths that might occur. PMID- 12635736 TI - The shared donor oocyte program: the advantages and insights it provides in determining etiologic factors of infertility. AB - A shared oocyte program allows a couple to achieve a pregnancy by in vitro fertilization (IVF) when this option may have been precluded because of financial considerations. It also allows recipients to save money since they do not have to pay a donor fee. The shared program offers the availability of a larger pool of oocytes thus reducing waiting times for recipients. The donors are a highly motivated group since they are working for their own infertility problem and are therefore very cooperative. The shared oocyte program is a very valuable research tool because one can evaluate under different circumstances the outcome of the same pool of oocytes being fertilized by different sperm and the subsequent embryos formed being transferred into two different women under different circumstances. PMID- 12635735 TI - Survival of Staphylococcus aureus ATCC 13565 in intermediate moisture foods is highly variable. AB - Staphylococcus aureus is a gram-positive, enterotoxin-producing coccus. It is a hardy organism and known to survive over a wide range of water activities, pH values, and temperatures. The objective of this study was to model the survival or gradual inactivation of S. aureus ATCC 13565 in intermediate moisture foods (IMFs). Various initial concentrations (approximately 10(1), 10(2), 10(3), and 10(4) CFU/g) were used to inoculate three different IMFs (beefsteak, bread, and chicken pockets). Viable counts were determined up to 60 days using tryptic soy agar. Inoculum size did not influence the survival or gradual inactivation of S. aureus in these foods. The rate of change (increase or decrease) in log CFU/day was calculated for every consecutive pair of data points and by linear regression for each inactivation curve. Both consecutive pair and linear regression rates of change were fit to logistic distributions (with parameters alpha and beta) for each food. Based on the distribution parameters, survival or gradual inactivation of S. aureus was predicted by computer simulation. The simulations indicated an overall decline in S. aureus population over time, although a small fraction of samples in the consecutive pair simulation showed a slight population increase even after 60 days, consistent with the observed data. Simulation results were compared to predictions from other computer models. The models of Stewart et al., were fail-safe, predicting the possibility of significant growth only after > 3,000 days. The USDA pathogen modeling program predictions were found to be fail dangerous, predicting declines at least four times faster than observed. PMID- 12635734 TI - Impact of microbial ecology of meat and poultry products on predictions from exposure assessment scenarios for refrigerated storage. AB - A novel extension of traditional growth models for exposure assessment of food borne microbial pathogens was developed to address the complex interactions of competing microbial populations in foods. Scenarios were designed for baseline refrigeration and mild abuse of servings of chicken broiler and ground beef Our approach employed high-quality data for microbiology of foods at production, refrigerated storage temperatures, and growth kinetics of microbial populations in culture media. Simple parallel models were developed for exponential growth of multiple pathogens and the abundant and ubiquitous nonpathogenic indigenous microbiota. Monte Carlo simulations were run for unconstrained growth and growth with the density-dependent constraint based on the "Jameson effect," inhibition of pathogen growth when the indigenous microbiota reached 10(9) counts per serving. The modes for unconstrained growth of the indigenous microbiota were 10(8), 10(10), and 10(11) counts per serving for chicken broilers, and 10(7), 10(9) and 10(11) counts per serving for ground beef at respective sites for backroom, meat case, and home refrigeration. Contamination rates and likelihoods of reaching temperatures supporting growth of the pathogens in the baseline refrigeration scenario were rare events. The unconstrained exponential growth models appeared to overestimate L. monocytogenes growth maxima for the baseline refrigeration scenario by 1500-7233% (10(6)-10(7) counts/serving) when the inhibitory effects of the indigenous microbiota are ignored. The extreme tails of the distributions for the constrained models appeared to overestimate growth maxima 110% (10(4)-10(5) counts/serving) for Salmonella spp. and 108% (6 x 10(3) counts/serving) for E. coli O157:H7 relative to the extremes of the unconstrained models. The approach of incorporating parallel models for pathogens and the indigenous microbiota into exposure assessment modeling motivates the design of validation studies to test the modeling assumptions, consistent with the analytical-deliberative process of risk analysis. PMID- 12635737 TI - Role of ultrasound evaluation of nuchal translucency in prenatal diagnosis. AB - BACKGROUND: Nuchal translucency (NT) is the ultrasonographic pattern of the accumulation of subcutaneous fluid (> or = 3 mm) behind the fetal neck. The measurement of NT thickness by ultrasound examination at 11-14 weeks of gestation has been associated with maternal age and to be an effective screening tool for trisomy 21; with an invasive method rate of 5%, about 75% of trisomical pregnancies can be identified. With the association of some biochemical markers like maternal serum free beta-human chorionic gonadotropin (beta-hCG) and pregnancy-associated plasma protein-A (PAPP-A) to ultrasonography at 11-14 weeks, it is possible to identify about 90% of chromosomal abnormalities. An increase of NT also allows us to identify most other chromosomal abnormalities, a large number of major cardiac defects, skeletal dysplasias, and genetic syndromes. In monochorial twins the discordance in the measurement of NT represents an early sign of twin-to-twin transfusion syndrome (TTTS). METHODS: The objective of our study was to assess the detection of fetal structural defects with an ultrasound scan at 11-14 weeks of gestation. We submitted 3,157 pregnant women to a routine ultrasound examination at 11-14 weeks. The patients were then submitted to ultrasound scan in the second or third trimester of pregnancy. An isolated increased NT thickness was not considered an abnormality, but these patients, nonetheless, were submitted to an early echocardiographic evaluation. Fetal structural abnormalities were classified as major or minor and of early or late onset. RESULTS: A detection rate of 4.3% (135 cases) of abnormalities was found and 22.6% of these (30 cases) were diagnosed by ultrasound scan at 11-14 weeks, including seven cardiac defects associated with increased NT. The antenatal ultrasound detection rate was 73.5%, and 33.2% were diagnosed in the first trimester assessment. A rate of 76.8% of the major defects were diagnosed by the prenatal scan and 35.8% by the scan at 11-14 weeks. Fetal structural defects with the ultrasonography at 11-14 weeks were diagnosed in about 24.3% of the cases, therefore, a second trimester abnormality is important in routine antenatal care to increase the prenatal assessment of fetal anomalies. CONCLUSIONS: As for the introduction of every new technique in routine clinical practice, the operators who perform the ultrasound scan at 11-14 weeks should be submitted to adequate training and to strict quality control. PMID- 12635738 TI - Nucleated red blood cell counts in growth-restricted neonates with absent or reversed-end-diastolic umbilical artery velocity. AB - OBJECTIVE: To determine the effect of increasing circulatory impairment in fetuses on the neonatal nucleated red blood cell count. METHODS: Seventy-seven fetuses with suspected intrauterine growth restriction (abdominal circumference < 5th percentile), subsequent birth weight < 10th percentile for gestational age and abnormal umbilical artery Doppler flow velocity were enrolled in the study. Fetuses were assigned to the following groups on the basis of the last Doppler examination before delivery: group 1, umbilical artery S/D ratio > 2 SD above the gestational age mean, group 2, absent end-diastolic velocity in the umbilical artery, group 3, reversed end-diastolic velocity in the umbilical artery. Neonatal nucleated red blood cells were obtained from the umbilical artery within 1 min after delivery with follow-up examinations until the nucleated red blood cell count was < 5/100 white blood cells. Perinatal and neonatal data were recorded from the charts. RESULTS: Significantly higher nucleated red blood cell counts [median 144, range 9-964] with lowest birth weights [mean 932 g, range 530 g-2060 g] were detected in group 3 and in group 2 [median 65, range 2-720; mean 1049 g, range 630 g-2110 g] compared to group 1 [median 22, range 2-201; mean 1565 g, range 860 g-2780 g, p < 0.001, respectively]. The persistence of nucleated red blood cells in the neonatal circulation was prolonged in group 3 [median 5.2, range 1-13 days] compared to group 2 [median 3.9, range 1-81 and group 1 [median 1, range 0-2]. Neonates in group 3 also had decreased platelet counts, hematocrit and hemoglobin values. Cord blood gases showed a significant decrease of arterial and venous pH values in groups 2 and 3 which was accompagnied by a significant decrease in base excess values. Four neonatal deaths, three of them in the reversed-end-diastolic velocity and one in the absent end-diastolic velocity group, occurred. Stepwise regression demonstrated that absent or reversed end-diastolic flow velocities, birth weight, gestational age and arterial base excess contribute significantly to the elevation of the nucleated red blood cell counts. CONCLUSIONS: Increased nucleated red blood cell counts at birth and their longer persistance in the neonatal circulation are associated with antenatal abnormal umbilical artery Doppler studies in growth restricted fetuses. The nucleated red blood cell count might therefore become an additional valuable tool in the diagnosis of growth restricted fetuses who are metabolically compromised. PMID- 12635739 TI - Tension-free vaginal tape procedure for treatment of USI: subjective and objective efficacy evaluation. AB - Urinary incontinence is a frequent functional disease affecting 5% to 25% of women. It is commonly believed that urinary stress incontinence (USI) is caused by anatomical defects, thus it can be treated by restoring anatomy. More than 100 surgical techniques have been proposed for genuine stress incontinence treatment. The tension free vaginal tape procedure (TVT) for the treatment of genuine IUS in women was first reported in 1996 by Ulmsten et al. The purpose of this study was to evaluate objective and subjective efficacy of the TVT procedure in different types of female urinary incontinence. From February 2000 to December 2001, 52 patients with urinary incontinence underwent the TVT procedure in the Obstetric and Gynecology Department of Avezzano Hospital, Italy. No difference in the cure rates was found between patients undergoing the TVT alone or associated with anterior and/or posterior colporrhaphy. In accordance with many authors we can affirm that the TVT sling technique is easy to use, the time for surgery is relatively short, and the procedure is free of complications and provides a satisfactory outcome. PMID- 12635740 TI - Ovine fetal growth curves in twin pregnancy: ultrasonographic assessment. AB - PURPOSE OF INVESTIGATION: The fetal-lamb model is a fundamental tool for developing clinical applications for the treatment of human fetal pathology (e.g., open fetal surgery). Accurate estimation of gestational age is important to avoid size-related problems during surgery. METHODS: To evaluate the effect of twinning on ovine fetal growth, we followed seven twin pregnancies in Comiso ewes from mating through parturition. Fetal growth indexes (muzzle-occipital and mean abdominal diameters, abdominal circumference, femoral and humeral length) were measured weekly using perimammary ultrasonography and analyzed with a linear quadratic regression model based on natural logarithms of each parameter and fetal age. RESULTS: The model explained > 90% of the variability observed, with determination coefficients of 95% (femoral length, abdominal circumference), 94% (abdominal diameter, humeral length), and 89% (muzzle-occipital diameter). CONCLUSION: Mean birth weight was lower than that reported for singleton fetal lambs, as it is in bigeminal pregnancies in humans, despite the uterine and placental differences between these two species. With the exception of slightly earlier growth deceleration, curves for head and long-bone growth resembled those for singleton ovine fetuses. Ovine fetal growth patterns (like those of humans) in singleton and twin pregnancies are similar. PMID- 12635742 TI - Thromboprophylaxis throughout pregnancy in women with previous history of recurrent miscarriages of unknown aetiology. AB - The purpose of this prospective preliminary clinical study was to assess the efficacy of thromboprophylaxis throughout pregnancy in women with a history of unexplained first trimester recurrent miscarriages. From the 53 patients originally assigned to the study 15 were excluded. The remaining 38 were treated with low molecular weight heparin (LMWH-natroparine calcium 0.3 ml twice daily) and low dose aspirin from the day the fetal heart motion was detected until the 37th week or earlier at the onset of premature labor. Among the patients treated (n = 38) thrombophilia screening was positive in 16 patients and in the remaining 22 no causative factor was detected. The overall success rate (viable pregnancy > or = 24 weeks) was 92.2% with no significant difference between patients with positive or negative thrombophilia screening. The most significant complications were: preeclampsia (21%), IUGR (26%), placenta abruptio (5.2%), injection site haematoma (44%) and skin reaction (15.7%). No abnormal bleeding was observed during vaginal or caesarean section. The results of this study suggest that thromboprophylaxis during pregnancy, which has already been successfully tried in patients with recurrent miscarriages with a causative factor, may be similarly effective in patients with such a pregnancy complication but of unknown aetiology. PMID- 12635741 TI - Can human fetal cortical brain tissue transplant (up to 20 weeks) sustain its metabolic and oxygen requirements in a heterotopic site outside the brain? A study of 12 volunteers with Parkinson's disease. AB - BACKGROUND: Neural and stem cell transplantation is emerging as a potential treatment for neurodegenerative diseases from Parkinson's to Huntington's disease. Stereotactic placement of dopaminergic neurons in the caudate-putamen (striatum), is being attempted in centers of excellence and has proved to be beneficial. Basic research using cell transplantation indicates that structural development mechanisms seen in immature brains, i.e., fetal brains, can also function in the adult brain. The adult brain consumes 15% of the resting cardiac output for its metabolic needs. While most human tissues can sustain an anaerobic assault for a few minutes up to 30 minutes, a sudden total lack of oxygen supply to the brain cells in an adult will render the person unconscious within five to ten seconds. Our team has been working on the problem of human fetal tissue response to antigenic assault for the last two decades. In the present series, 12 patients with prolonged histories of Parkinsonism, who were not responding to anti-Parkinsonian drugs, and could not afford costly stereotactic surgery or deep brain stimulation and other modalities of recent Parkinson's disease treatment, were enrolled in the study. MATERIALS AND METHOD: After obtaining proper informed consents from the patients or their guardians and from the multidisciplinary ethical committee, the patients, varying in age from 45 to 75 years and suffering for many years with Parkinsonism, were enrolled in the heterotopic brain tissue transplant programme. We followed standard antiseptic, aseptic and premedication protocols, after selecting a proposed site of transplantation of the brain in the axillary fold of the skin, under local infiltration anaesthesia. In an adjacent OR, a fetus was collected from a consenting patient undergoing hysterotomy and ligation (before 20 weeks), under general anaesthesia. Within a minute of hysterotomy, the fetal brain tissue was dissected, and under the guidance of the operative microscope, 1 g of fetal cortical brain tissue was dissected and weighed in an electronic machine. The tissue was collected from around 1 cm of the frontal opercula of the developing human fetal brain and grafted in the already dissected and prepared subcutaneous site in the axilla and the skin was closed. Hematological parameters (Hgb; total count, Tc; differential count, Dc; erythocyte sedimentation rate, ESR) were estimated sequentially up to one month. A small portion of the transplanted tissue was retrieved after one to two months, and a serial histological study was done along with a clinical assessment of the disease condition as per the specifications of the Unified Parkinson's Disease Rating Scale. The results were matched with the pre-transplant ratings of the individual cases. Presenting dyskinesia was also rated (0-4), on the basis of objective criteria assessment like walking, putting on a coat, lifting a cup to drink, etc. RESULTS AND ANALYSIS: Initially 30 patients suffering from advanced Parkinson's disease (PD) were approached after getting the necessary clearance from the institutional multidisciplinary ethical committee; however, we have been able to arrange transplantation in only 12 cases so far. These patients were evaluated at the pre- and one month post-transplant period by the Unified Parkinson's Disease Rating Scale (0-108) and the minimum score was 40 in the motor portion of the unified scale at the pre-transplant state. Evaluation of the patients after one month revealed mild improvement of the pre-transplant scoring (up to 33.3%) in 41.6% of the cases, and moderate improvement (up to 66.6%) in another 41.6% of the cases. While 16.8% of the cases did not show any improvement from the basal score, i.e., the pre-transplant score, there was a definite sense of well being and rise in weight (2-4 pounds) noted in each case and there was also a reduction of the L-Dopa dosage in 75% of the cases. There was also a 58.3% improvement in the bradykinesia scoring from the pre-transplant level. What is intriguing is the survival, growth and proliferation of the grafted fetal brain tissue in the HLA- and sex-randomized adult axilla without any immunosuppressive support. Not a single histological study of the fetal brain tissues after removal from the axilla showed any signs of graft vs. host or inflammatory reaction (Figures 1-9) but there were features of growth of the transplanted cortical brain tissue along with its different components like neurogenesis, gliogenesis, early neovascularisation and angiogenesis, etc. There was also no systemic leucocytosis or lymphocytosis. DISCUSSION AND CONCLUSION: Histological evidence at the transplanted tissue site suggests that fetal cortical brain tissue can sustain life in sex-randomized, HLA-randomized adult hosts, without the support of immuno-suppressive drugs and the tacit support of the blood-CSF and blood brain barrier and other specific requirements of adult brain cells in the skull. Whether the clinical improvement in PD is transient or long lasting is presently under investigation along with basic questions like, is it due to transplanted fetal dopaminergic or non-dopaminergic neurons or is it the growth factors and the cytokine mediated hitherto unknown reactions causing the clinical improvement. PMID- 12635743 TI - Reproductive outcome after laparoscopic treatment of endometriosis. AB - PURPOSE OF INVESTIGATION: To evaluate the pregnancy rate after laparoscopic treatment of endometriosis. PATIENTS AND METHODS: 47 infertile women undergoing laparoscopic treatment of endometriosis. RESULTS: rAFS stage was as follows: 11% of patients had Stage I, 11% Stage II, 53.3% Stage III and 24.4% Stage IV. The mean duration of follow-up was 48.5 +/- 18.44 months. The overall pregnancy rate was 64.4%. Eighteen out of 26 women (69%) became pregnant within six months after laparoscopy, 23% at 12 months, 11% within 24 months, and 11% after two years (p < 0.01). Adnexal adhesions and tubal status significantly affected the pregnancy rate. No differences were found regarding the stage of disease and the presence of ovarian endometriomas. CONCLUSION: Laparoscopic treatment of endometriosis enhances fertility and the pregnancy rate is highest within the first six months after surgery. Adnexal adhesions and tubal conditions influence the reproductive outcome. PMID- 12635744 TI - The efficacy of immediate post-abortion intrauterine device insertion. AB - Ovulation returns soon after abortion, with half of the women ovulating by 21 days after the procedure. Immediate post-abortion insertion provides immediate contraceptive protection. Clinicians have feared that rates of uterine perforation due to intrauterine devices (IUDs) and expulsion of IUDs may be higher in the post-abortal uterus due to its softness and enlargement. This study was carried out to assess the safety and efficacy of IUD insertion immediately after induced abortion compared with insertion during the first menstrual cycle after abortion. All women were counseled about a method of contraception, particularly copper T-380A, and divided into two groups: Group 1 included 226 women who preferred immediate IUD insertion, and Group 2 included 100 women who opted for interval-IUD insertion during the first menstrual cycle after abortion. All women were followed-up at two and eight weeks after insertion of the IUDs. Severe bleeding was observed 10.62% in Group 1 and 6% in Group 2. Expulsion rate was 7.08% and 8% in Groups 1 and 2, respectively, which was not statistically significant. Pelvic inflammatory disease (PID) was observed in one case in Group 1. However uterine perforation was not noted in either group. In conclusion, insertion of an IUD immediately after an induced abortion is safe and if a woman asks for a modern contraceptive method it could be offered. PMID- 12635745 TI - Laparoscopic-ultraminilaparotomic myomectomy (LUM)-laparoscopic ultraminilaparotomic embolized myomectomy (LUEM). Surgical techniques. AB - Laparoscopic myomectomy has been performed for more than 20 years. More recently other techniques such as laparoscopically assisted vaginal myomectomy (LAVM) and laparoscopically assisted myomectomy (LAM) have also been introduced. Laparoscopic-ultraminilaparotomic myomectomy (LUM) and laparoscopic ultraminilaparotomic embolized myomectomy (LUEM), a new surgical technique which integrates laparoscopy and ultraminilaparotomy and embolization, has been created by our group and has been found to be superior to the conventional laparoscopy for the treatment of uterine fibromas especially in large myomas > 9 cm. This technique allows us to apply a suture on the uterine incision totally similar to the suture of a conventional laparotomy, by using the small breach of the cutaneous incision of the uterine morcellator (25 mm) and drawing the uterus below this abdominal aperture. This kind of surgical procedure is associated with presurgical embolization, that we call LUEM (laparoscopic ultraminilaparotomic embolized myomectomy) of the afferent vessels to the myoma in myomas with diameters equal or superior to 14 cm to avoid blood loss during the surgical procedure. Between June 1999 and March 2002, a total of 62 patients wishing to become pregnant were treated with this method. LUM allows us to increase the feasibility and safety of the operation, while assuring a better stability of the uterine suture and reduction of surgical time. LUEM has the advantages of LUM but permits in an absolutely hemostatic situation the application of laparoscopy in the surgery of large myomas superior to 14 cm. PMID- 12635746 TI - Should asymptomatic bacteriuria be screened in pregnancy? AB - The incidence of asymptomatic bacteriuria is reported as 2-14% during pregnancy. Fetal and maternal complications like acute pyelonephritis, hypertension, anemia, preterm labor, low-birth-weight infants and intrauterine growth retardation can be expected. The purpose of this study was to determine the incidence of asymptomatic bacteriuria during pregnancy and its relation to pregnancy complications. The study involved 270 pregnant women up to 32 gestational weeks during a 9-month period. At the initial visit, they were screened with urine culture in order to detect asymptomatic bacteriuria. A control group was formed in a retrospective manner from the first day of the study with 186 pregnant women who delivered in our clinic and who were not screened for asymptomatic bacteriuria. The incidence of asymptomatic bacteriuria was 9.31%. Escherichia coli accounted for 79%, which was the most frequent of the isolates. We observed recurrence and had to apply treatment again to 21.7% of the women. The sensitivity, specificity, positive predictive and negative predictive values of leucocyturia as a screening test for asymptomatic bacteriuria were 91.3%, 83.6%, 45.6% and 98.5%, respectively. We diagnosed preterm labor in six of 23 (26%) with asymptomatic bacteriuria and 16 in 163 (9.3%) women in the urine culture negative group. The ratio acute pyelonephritis in the group which was routinely screened and treated for asymtomatic bacteriuria was 0.5% while the prevalence was 2.1% in the nonscreened group. Considering the relatively high incidence of asymptomatic bacteriuria during pregnancy and the relevant complications, we propose to screen and treat asymptomatic bacteriuria routinely in all pregnant women. PMID- 12635747 TI - A comparative study of the effect of ovarian stimulation protocols with different gonadotropin preparations on the biological and clinical parameters of the outcome of intracytoplasmic sperm injection. AB - Intracytoplasmic sperm injection (ICSI) is widely employed today in cases of severe male factor infertility. This technique requires denuding the oocytes from the surrounding granulosa cells prior to sperm injection. One can thus assess oocyte maturity more accurately and can study the effects of various ovarian stimulation protocols on egg maturation and the rest of the parameters of the outcome of ICSI. The aim of the present study was to compare the outcome of ovarian stimulation using human menopausal gonadotropin (hMG) with that achieved by using highly purified follicle stimulating hormone (pFSH). The biological and clinical parameters of the outcome of ICSI in 99 subfertile couples were studied. Group A consisted of 46 patients to whom hMG was administered and Group B consisted of 53 patients to whom pFSH was employed for ovarian stimulation. The fertilization rate was significantly higher in the pFSH group but all other factors were similar, including the percentage of mature oocytes and pregnancy rate. The latter does not seem to be affected by the gonadotropin preparation employed for ovarian stimulation. This is very helpful for the physician to know since a gonadotropin with a lower cost can be employed and, in addition, shortage of some preparations of gonadotropins occurs frequently. PMID- 12635748 TI - Endometrial fluid accumulation during controlled ovarian stimulation for ICSI treatment. A report of three cases. AB - The occurrence of endometrial fluid accumulation was examined in patients undergoing ovarian stimulation in a program of intracytoplasmic sperm injection (ICSI), by vaginal ultrasound. Endometrial fluid accumulation was recorded in three cycles out of 124. In one case, the fluid was absorbed before embryo transfer (ET), but in the other cases it was present in ET. In these three cases, the endometrium had been evaluated as normal before ovarian stimulation. Fluid accumulation in the endometrial cavity possibly affects the implantation process negatively. Therefore, alternative options should be considered as cancellation of the embryo transfer and cryopreservation of embryos to be available in a subsequent mild stimulated cycle. PMID- 12635749 TI - Serous papillary cystic ovarian borderline tumor: case report. AB - A case of a 30-year-old patient with a serous papillary borderline tumor of the right ovary who underwent a simple mono-lateral salpingo-oophorectomy is presented. Conservative surgery allowed a subsequent pregnancy with spontaneous delivery 27 months later. PMID- 12635751 TI - Two consecutive ectopic pregnancies after in-vitro fertilization and embryo transfer. Case report. AB - Ectopic pregnancy is a known complication of in vitro fertilization and embryo transfer (IVF/ET). The overall incidence of ectopic pregnancy after IVF is 4.4 5.8% of clinical pregnancies. The risk factors associated with ectopic pregnancies are complex. We present a patient with two consecutive ectopic pregnancies after IVF/ET. PMID- 12635750 TI - Complementary therapy for severe Rh-alloimmunization. AB - PURPOSE OF INVESTIGATION: This report describes successful treatment, using invasive and noninvasive techniques, of a 36-year-old woman (gravida 10, para 0) referred to our center at 13 weeks' gestation for severe Rh alloimmunization. Pre pregnancy indirect Coombs titers ranged from 1:1024-2048. All nine past pregnancies (conceived with three different partners) had ended in abortion, intrauterine death or neonatal death METHODS: The patient was treated with a single session of plasmapheresis (week 14) immediately followed by five days of immunoglobulin therapy and immunosuppressive therapy based on azathioprine and prednisone (weeks 15-22). Seven fetal transfusions (one intraperitoneal, six intravascular) were performed beginning at 16 weeks. RESULTS: The pregnancy, which was characterized by insulin-dependent gestational diabetes, spontaneously resolving polyhydramnios and peak indirect Coombs titers of 1:65,536, ended at 27 weeks with cesarean section delivery of a viable female weighing 1,000 g. In spite of numerous neonatal complications, the child is physically well at age 3, with normal intellectual and psychomotor development. CONCLUSION: In light of the negative outcomes of the patient's nine past pregnancies, our experience suggests that the early initiation of an integrated approach based on noninvasive and invasive techniques can play a potentially decisive role in the management of severe Rh-alloimmunization. PMID- 12635752 TI - Successful treatment with carbimazole of a hyperthyroid pregnancy with hepatic impairment after propylthiouracil administration: a case report. AB - We report the case of a 27-year-old woman with hyperthyroidism during pregnancy. Antithyroid treatment with propylthiouracil (PTU) resulted in elevated hepatic enzymes and after the 12th week of pregnancy treatment was changed to carbimazole (CBZ). The remaining pregnancy, delivery and follow-up period were uneventful for the mother and her offspring. Antithyroid treatment during pregnancy should allow the use not only of PTU but also of CBZ and methimazole. PMID- 12635753 TI - Cleft lip and/or palate: searching for aetiological factors. PMID- 12635754 TI - Cleft lip and palate: the Jos experience. AB - OBJECTIVE: To determine the pattern of occurrence of cleft lip/palate and the factors that may have influenced treatment outcome. DESIGN: Descriptive Study. SETTING: Jos University Teaching Hospital, Jos, Nigeria. SUBJECT: This study included 107 consecutive patients with cleft lip/palate managed between January 1991 and June 1997. MAIN OUTCOME MEASURES: The pattern of occurrence of cleft lip/palate, the peculiarities of the malformation in this environment as well as factors that influenced treatment outcome. RESULTS: The 107 patients were aged between one day and twenty-six years at presentation. There were three adults aged between 18 and 26 years with a mean of 22.3 years and 104 children with a mean age of 9.5 months. The male/female ratio was 1. 1:1, the anomaly was 2.4 times commoner on the left and the cleft lip alone was the most frequent mode of presentation (52%). The incidence was higher in the 3rd and 4th siblings. In 13% of these patients, there were other associated congenital anomalies such as Van der Woude's Syndrome, Down's Syndrome and congenital heart disease. Complications were noted in 16(14.9%) patients and these included dehiscence after repair in 8(7.5%) patients, oronasal fistulae in 2(1.9%), nasal speech in 4(3.7%) and hypertrophic scar in 2(1.9%). Eighty nine percent of these repairs were satisfactory to the parents of the affected children. All the adult patients were satisfied with their repair. CONCLUSION: The low incidence of this anomaly in our environment may be due to underreporting in the rural areas. Public enlightenment programs should help improve early presentation. PMID- 12635755 TI - Prevalence of asthma, allergic rhinitis and dermatitis in primary school children in Uasin Gishu district, Kenya. AB - OBJECTIVE: To establish the relative increase in the prevalence of asthma, allergic rhinitis and eczema in primary school children aged 13-14 years over a six year interval. DESIGN: Cross sectional comparative study. SETTING: Primary schools in three rural divisions at Uasin Gishu district in the Rift Valley Province of Kenya. METHODS: Three thousand two hundred and fifty eight children aged 13-14 years from seventy two primary schools in Uasin Gishu district were studied using the International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire. All children in the selected schools in this age range whose parents gave consent were included. RESULTS: There were 47.4% males and 52.6% females as compared to 48.7% and 51.3% respectively in 1995. The cumulative prevalence for wheezing, rhinitis, itchy eyes and dermatitis was 23.6%, 43%, 24.1% and 28.5% respectively which were higher than for 1995 which were 21.2%, 32.4%, 11.8% and 13.8% respectively (p = 0.001). The period prevalence rates for wheezing, rhinitis and eczema were 13.8%, 31.4% and 21.3% respectively as compared to 10.2%, 25.3% and 14.4% respectively for 1995 (p = 0.001). The prevalence of asthma, allergic rhinitis and eczema was 12.6%, 38.6% and 28.5% respectively in 2001 compared to 6.6%, 14.9% and 13.9% respectively in 1995 (p = 0.001). CONCLUSION: There was a significant increase in the prevalence of asthma, allergic rhinitis and eczema in children in the study population over the last six years. PMID- 12635756 TI - Headache associated disability in medical students at the Kenyatta National Hospital, Nairobi. AB - OBJECTIVE: To study headache associated disability in a group of medical students at the Kenyatta National Hospital. STUDY DESIGN: Cross sectional survey. RESULTS: Between October 1994 and January 1995 we conducted a survey on headache characteristics on medical students at both the Kenya Medical Training Centre and the Medical School of the University of Nairobi. Six hundred and twenty-five (87%) of the 711 students surveyed admitted having had at least one episode of headache in the last six months. Using the International headache society (IHS) case criteria 314 students (50%) had tension type headache, 240 (38%) migraine headache and 71(12%) unclassified headache. Eighty-six percent of the students with headache had their working ability disturbed to various degrees. Eighty-five percent of the students reported that their social activities were interfered with by headache. Migraine headaches had the greatest impact on both the working and social activities at a p-value of 0.0005 and 0.0004 respectively. One hundred and forty-one students (23.6%) had missed at least one day of work or school in the last one-year as a direct result of the headache. There was an association between headache severity with working ability and social effect. There was no association between the days students missed work or classes with the severity of the headache. No gender difference was found in the headache associated disability. CONCLUSION: Headache is a prevalent condition with disability both in working and social activities. PMID- 12635757 TI - Child care practices and nutritional status of children aged 0-2 years in Thika, Kenya. AB - OBJECTIVE: To assess time allocation for child care and the nutritional status of children aged 0-2 years. DESIGN: Cross sectional descriptive survey using a structured questionnaire and taking of anthropometric measurements to determine the nutritional status of children aged 0 to 2 years. In addition, two day (10 hour) observations were conducted in a subsample of households to assess time allocation for the main child care activities. SETTING: A low-income peri-urban section of Thika town (in Makongeni estate), Kenya. SUBJECTS: A random sample of 150 mothers and their 0-2 year old children. RESULTS: Mother's knowledge about child care influences the amount and type of care that is given to children. Time taken to perform various activities was also found to vary with the mother's education level, her occupation, number of children less than five years in the house and the child's age and birth order. Comparatively, children who were malnourished (stunted) had less time devoted to them for breastfeeding, food preparation and feeding. Although mothers were the primary caregivers, the responsibility of care giving was shared with other household members as well as with neighbours. CONCLUSION: The amount and type of care that a child receives is determined to a large extent by the mother and caregivers knowledge. PMID- 12635758 TI - Pain relief using paracervical block in patients undergoing manual vacuum aspiration of uterus. AB - OBJECTIVE: To evaluate pain relief using paracervical nerve block with 1% lignocaine injection in patients undergoing uterine evacuation by Manual Vacuum Aspiration (MVA) for the treatment of incomplete abortion. DESIGN: A randomized double blind clinical trial. SETTING: Marie Stopes Health Centre, Nairobi. METHODS: One hundred and forty two patients were recruited between September and October 1997. The intervention was random assignment to the study group (paracervical block with 1% lignocaine) or the placebo group (paracervical block with sterile water for injection). Intra and post operative assessment of pain was made using McGills and facial expression scales. RESULTS: The untreated group experienced significantly more pain than the treated group, especially lower abdominal pain and backache. The pain was especially marked intraoperatively, less so 30 minutes post-operatively. CONCLUSION: Based on the findings of this study, any patient going for manual vacuum aspiration for the treatment of incomplete abortion should be given Paracervical block as it is cost effective, easy to perform and with less side effects. PMID- 12635759 TI - Risk factors for placenta praevia in southern Nigeria. AB - OBJECTIVE: To determine the risk factors for placenta praevia in Ile-Ife, southern Nigeria. DESIGN: A prospective case control study. SETTING: A tertiary center--Obafemi Awolowo University Teaching Hospital, Ile-Ife, southern Nigeria. SUBJECTS: One hundred and thirty six patients with confirmed placenta praevia constituted the cases. Controls consisted of one hundred and thirty six patients who delivered at term immediately after each indexed case and did not have placenta praevia. RESULTS: Cases and controls were similar in terms of twin deliveries (P = 0.72) and past history of uterine surgery (P = 0.47). After adjusting for confounders, factors associated with risk of placenta praevia were history of retained placenta [OR = 6.7(95% CI 1.2-36.6)], previous caesarean section [OR = 4.7, (95% CI 1.9-11.4)], previous abortion [OR = 2.9 (95% CI 1.1 5.1)], grand multiparity [OR = 2.1 (95% CI 1.6-7.1)] and age over 35 years [OR = 1.4 (95% CI 1.2-6.6)]. CONCLUSIONS: From our study, the risk factors for placenta praevia are a history of retained placenta, previous caesarean section, previous abortion, grand multiparity and maternal age over 35 years. PMID- 12635760 TI - Morbidity and outcome of low birthweight babies of adolescent mothers at Kenyatta National Hospital, Nairobi. AB - OBJECTIVE: To compare the morbidity and outcome of low birthweight babies (birthweight < 2000 gm) of adolescent (age < 20 years) and older mothers. DESIGN: Cross sectional descriptive study. SETTING: The newborn Unit of the Kenyatta National Hospital. MAIN OUTCOME MEASURES: All babies weighing less than 2000 gm at birth whose mothers consented to the study had their gestational age verified using the Dubowitz scoring system. They were then followed up by daily clinical assessment until discharge, death or up to one month in the ward. The babies were divided into two groups according to their mother's age and then compared with respect to episodes of illness, duration of hospital stay, and overall outcome. RESULTS: One hundred and forty two babies were studied. Of these, 64 were born to adolescent mothers. Babies of the adolescent mothers tended to be more premature (p = 0.0174), be lower in weight (p = 0.0078), had more occurrences of respiratory distress and anaemia (probably reflecting their increased prematurity) and had frequent multiple morbidity events They also had longer hospital stay and they were more likely to die (57.7% compared to 42.3% of babies of older mothers). CONCLUSION: Low birthweight babies of the adolescent mothers were found to be more likely to have increased morbidity and adverse outcome compared to similar babies of older mothers. PMID- 12635761 TI - Low birthweight babies: socio-demographic and obstetric characteristics of adolescent mothers at Kenyatta National Hospital, Nairobi. AB - OBJECTIVE: To compare some socio-demographic and obstetric factors between adolescent mothers (aged below 20 years) and older mothers of low birthweight (birthweight < 2000 gm) babies. DESIGN: Cross sectional descriptive study. SETTING: The Newborn Unit of the Kenyatta National Hospital. RESULTS: Sixty nine adolescent mothers and 73 older mothers were studied. Adolescent mothers were more likely to be unmarried (p = 0.0001) have less formal education (p < 0.0001) be unemployed and be primigravida (76.5% compared to 36% of older mothers). Although the obstetric factors of antenatal clinic attendance, premature rupture of the membranes, pre-eclamptic toxaemia, infections and interventronal delivery tended to be more frequent among the adolescent mothers, non of these differences were significant probably due to the small numbers of patients studied. CONCLUSION: This study does suggest mothers of very low birthweight babies tend to have unfavourable socio-demographic and obstetric factors like being single parents having less formal education, being unemployed and having obstetric risks for poor pregnancy outcome. PMID- 12635763 TI - Cryptosporidiosis among animal handlers and their livestock in Basrah, Iraq. AB - OBJECTIVE: To investigate the prevalence of cryptosporidiosis among groups at risk (animal handlers) and among domestic animals. DESIGN: Comparative study with zoonotic aspect. METHOD: Stool samples were collected from 60 animal handlers, 175 non-animal handlers and 198 domestic animals (60 cows, 45 sheep, 45 goats, 25 horses and 23 camels). Direct smear method and then formalin-ether sedimentation method were carried out for stool samples to detect intestinal parasites. Faecal smears were prepared from the sediment and stained by the modified Ziehl-Neelsen method for the recovery of red-pink oocysts of Cryptosporidium. RESULTS: Out of the 60 animal handlers, 30 (50%) were found to be positive for intestinal parasites compared to 26 (14.8%) of non-animal handlers (P < 0.01). Cryptosporidium oocysts were found to be excreted by three (5%) animal handlers and two (1.14%) of the non-animal handlers (P > 0.05). Cryptosporidiosis was also diagnosed in 20%, 13.3%, 17.7% and 12% of cattle, sheep, goats and horses respectively. No single positive case was detected among the examined camels. CONCLUSION: Veterinarians, butchers and breeders should be aware of the disease among farm animals in order to avoid great losses and to prevent its transmission to humans. PMID- 12635762 TI - Prevalence of tardive dyskinesia among psychiatric in-patients at Mathari Hospital, Nairobi. AB - OBJECTIVE: To determine the prevalence of tardive dyskinesia among psychiatric in patients. DESIGN: A cross-sectional survey. SETTING: Mathari Hospital, Nairobi, the main psychiatric referral hospital in Kenya. SUBJECTS: Two hundred and two randomly selected in-patients seen in the hospital between January and April 2000. RESULTS: The prevalence of tardive dyskinesia was 11.9%. Neither the psychiatric diagnosis nor the sex was significantly associated with tardive dyskinesia. The antipsychotic dosage was also not associated with tardive dyskinesia but an increase in age was significantly associated with the abnormal movements. CONCLUSION: The prevalence rate of tardive dyskinesia among patients at Mathari Hospital is much lower than that found in western countries but similar to that from Asian studies. These findings indicate the possibility of racial differences in the aetiology of TD. Prospective cross-racial studies are necessary to confirm these findings. PMID- 12635764 TI - Persistent hyperinsulinaemic hypoglycaemia of infancy: case report. AB - Hyperinsulinism, although rare, is the most common cause of persistent hyperinsulinaemic hypoglycaemia in infancy. Because of persistent hypoglycaemia, serious difficulties are encountered in the long term management of this condition. A male neonate, after an uncomplicated full-term pregnancy, had been admitted to another hospital with convulsions on the third post-natal day. Meningitis had been suspected at that time and treated with phenobarbital and he had been discharged from the hospital. At three-months old he was referred to our department for persistent convulsions and lethargy. His parents were of 1st degree consanguinity. His blood glucose level was found to be 24 mg/dl (1.33 mmol/L). Because of the dangerously high insulin level during hypoglycaemia (insulin/glucose > 0.3), the absence of ketonuria, and the need for a high dose of glucose infusion (> 15 mg/kg/min) to achieve normoglycaemia and a glycaemic response to glucagon despite the hypoglycaemia, a diagnosis of persistent hyperinsulinaemic hypoglycaemia of infancy was made. Since maximal doses of prednisone, glucagon, diazoxide, octreotide and high infusion of glucose were ineffective in achieving normoglycaemia, a subtotal (80%) pancreatectomy was done. Postoperatively intermittent hypoglycaemic episodes continued. These were controlled with low doses of octreotide. Histology revealed diffuse adenomatous hyperplasia (nesidoblastosis). The boy is now in the sixth post-operative month and developing normally. PMID- 12635765 TI - Leiomyoma of the urinary bladder: case report. AB - A case of leiomyoma of urinary bladder, a rare benign tumour, is presented. The patient was a 42 year old female who presented with dysuria and frequency of micturition. The radiological features, diagnosis and management are discussed and the literature on this subject is briefly reviewed. PMID- 12635766 TI - Vibrio cholerae 01 strain resistant to vibriostatic compound 0/129 isolated from cholera cases in Kilifi, Kenya. PMID- 12635767 TI - Synthesis and characterization of poly(L-lysine)-g-poly(D,L-lactic-co-glycolic acid) biodegradable micelles. AB - A series of amphipathic graft copolymers composed of poly(L-lysine) (PLL) as the cationic polymer backbone and biodegradable poly(D,L-lactic-co-glycolic acid) (PLGA) as the grafting chains were synthesized and characterized. The terminal group of PLGA was activated and chemically conjugated to the primary epsilon amine groups of PLL to produce PLL-g-PLGA copolymers. PLL-g-PLGA formed a self assembling micelle structure in aqueous solution. The micelle size ranged from 69.4 to 149.6 nm in diameter, depending on the grafting percentage of PLGA. Upon increasing the number of PLGA chains grafted onto the PLL backbone, the size of the micelles gradually decreased, at the same time lowering their critical micelle concentration. The micelles were individually separated and had a spherical geometry, as observed by atomic force microscopy (AFM). These PLL-g PLGA copolymers can be applied as cell adhesive surface coating materials for biodegradable tissue engineering scaffolds and can be used as non-viral DNA carriers for gene therapy. PMID- 12635768 TI - Surface modification of poly-L-lactic acid (PLLA) membrane by grafting acrylamide: an effective way to improve cytocompatibility for chondrocytes. AB - Poly-L-lactic acid (PLLA) membranes were photo-oxidized in hydrogen peroxide solution under ultraviolet light (UV) to introduce hydroperoxide groups onto the PLLA membrane surfaces. The photo-oxidized membranes were then immersed in acrylamide (AAm) solution containing Fe2+ to graft polyacrylamide (PAAm) onto the PLLA membrane surfaces. The density of the hydroperoxide groups introduced on the PLLA membrane surfaces varied with the temperature and the photo-oxidization time. The occurrence of grafting was verified by X-ray photoelectron spectroscopy (XPS). The degree of grafting increased with the monomer concentration and the polymerization time. Water contact angle measurements showed that the wettability of the modified PLLA membranes had improved. Chondrocytes proliferated more rapidly and were more spread out on the modified membrane than on the control PLLA membrane, indicating that the PAAm-grafted PLLA membrane has better cytocompatibility for chondrocytes. PMID- 12635769 TI - Effects of the preparation methods of hydroxypropyl methylcellulose/polyacrylic acid blended films on drug release. AB - Hydroxypropyl methylcellulose (HPMC)/polyacrylic acid (PAA) blended films were prepared under different conditions (i.e. temperature, solvent, and cross-linking agent). The effects of hydrogen-bonding interactions or a chemical reaction between the two-component polymers in the blended films on drug release were studied. Two model drugs were used for comparison: a water-soluble drug, dl propranolol hydrochloride, and a lipophilic drug, ketoprofen. The H-bonding interaction of HPMC/PAA was found to be stronger in the blended films prepared from H2O than that from H2O/EtOH. However, the H-bonding effect between HPMC and PAA on drug release is indistinct. Chemical esterification of the carboxylic acid groups of PAA with the hydroxyl groups of HPMC was proposed from the solid-state NMR, ESCA, and FTIR studies when the blended films were dried at 110 degrees C. This chemical reaction also resulted in insolubilization of the blended films. PMID- 12635770 TI - In vitro and in vivo degradation studies of a novel linear copolymer of lactide and ethylphosphate. AB - Poly(lactide-co-ethylphosphate)s, a new class of linear phosphorus-containing copolymers made by chain-extending low-molecular-weight polylactide prepolymers with ethyl dichlorophosphate, were investigated for their in vitro and in vivo degradation mechanism and kinetics. Microspheres made from poly(lactide-co ethylphosphate) were studied under both accelerated and normal in vitro degradation conditions. Gel permeation chromatography (GPC), 1H- and 31P-NMR, weight loss measurements, and differential scanning calorimetry (DSC) techniques were used to characterize the change of molecular weight (M(w)), chemical composition, and glass transition temperature (T(g)) of the degrading polymers. The results indicated that the copolymers degraded in a two-stage fashion, with cleavage of the phosphate-lactide linkages contributing mostly to the initial more rapid degradation phase and cleavage of the lactide-lactide bonds being responsible for the slower latter stage degradation. The decrease in the copolymer M(w) was accompanied by a continuous mass loss. Results from the accelerated degradation studies confirmed that the copolymers degraded into various monomers of the copolymers, which were non-toxic and biocompatible. A two stage hydrolysis pathway was thus proposed to explain the degradation behavior of the copolymers. In vivo degradation studies performed in mice demonstrated a good in vitro and in vivo correlation for the degradation rates. In vivo clearance of the polymer was faster and without any lag phase. These copolymers are potentially advantageous for drug delivery and other biomedical applications where rapid clearance of the polymer carrier and repeated dosing capability are essential to the success of the treatment. PMID- 12635772 TI - Molecular weight distribution changes during degradation and release of PLGA nanoparticles containing epirubicin HCl. AB - The molecular dynamics of the degradation process of poly(lactic-co-glycolic acid) nanospheres were investigated during the degradation process usually observed when these polymers are used as controlled release carriers. The molecular weight distribution of PLGA samples was determined over a period of 32 days by accurately analyzing the molecular weight distribution of the polymer as a function of time as degradation progressed. The molecular weight distribution shifted gradually to lower average molecular weights over 32 days, with significantly smaller molecular weight components appearing at 8-12 days. In addition, the degradation of nanospheres containing epirubicin HCI was analyzed and increasing the amount of epirubicin from 1.7 to 3.4 to 6.7 wt% was found to hasten the degradation of the nanoparticles and subsequently affect the release behavior from these particles. This is believed to be the first time that such molecular dynamics have been presented for the degradation of PLGA nanoparticle formulations containing a drug for controlled delivery. PMID- 12635771 TI - Amphiphilic copolymers of epsilon-caprolactone and gamma-substituted epsilon caprolactone. Synthesis and functionalization of poly(D,L-lactide) nanoparticles. AB - Fully biodegradable and surface-functionalized poly(D,L-lactide) (PLA) nanoparticles have been prepared by a co-precipitation technique. Novel amphiphilic random copolyesters P(CL-co-gammaXCL) were synthesized by controlled copolymerization of epsilon-caprolactone and epsilon-caprolactone substituted in the gamma-position by a hydrophilic X group, where X is either a cationic pyridinium (gammaPyCL) or a non-ionic hydroxyl (gammaOHCL). Nanoparticles were prepared by co-precipitation of PLA with the P(CL-co-gammaXCL) copolyester from a DMSO solution. Small amounts of cationic P(CL-co-gammaPyCL) copolymers are needed to quantitatively form stable nanoparticles (ca. 10 mg/ 100 mg PLA), although larger amounts of non-ionic P(CL-co-gammaOHCL) copolymers are needed (> or = 12.5 mg/ 100 mg PLA). Copolymers with a low degree of polymerization (ca. 40) are more efficient stabilizers, probably because of faster migration towards the nanoparticle-water interface. The nanoparticle diameter decreases with the polymer concentration in DMSO, e.g. from ca. 160 nm (16 mg/ml) to ca. 100 nm (2 mg/ml) for PLA/P(CL-co-gammaPyCL) nanoparticles. Migration of the P(CL-co gammaXCL) copolyesters to the nanoparticle surface was confirmed by measurement of the zeta potential, i.e. ca. +65 mV for P(CL-co-gammaPCL) and -7 mV for P(CL co-gammaOHCL). The polyamphiphilic copolyesters stabilize PLA nanoparticles by electrostatic or steric repulsions, depending on whether they are charged or not. They also impart functionality and reactivity to the surface, which opens up new opportunities for labelling and targeting purposes. PMID- 12635773 TI - Tooth-by-tooth survival analysis of the first caries attack in different age cohorts and health centers in Finland. AB - The aims of this study were to test the hypothesis that existing patient records serve as a source of data for survival analysis of dental health and to develop a system for routinely conducted survival analysis of dental health from tooth eruption to first caries attack. Patient records for three cohorts were collected from four towns in Finland: Oulu, Turku, Kuopio, and Kemi. The study population comprised 2103 children born and living in these towns. The Kaplan-Meier curves for dental caries were drawn separately for each tooth and for the maxilla and mandible (for each age cohort). Placements of the first restoration of all teeth in each age cohort were investigated. The 1960 cohort had an extremely high post eruptive morbidity in all teeth. The 1970 cohort had relatively high immediate post-eruptive morbidity in both molars, but much lower than that of the 1960 cohort. The filling increment rates for second molars were lower than those of the first molars in the 1970 cohort. Characteristic of the 1980 cohort was a rapid increase in the caries increment of the premolars and second molars 4 years after eruption. The results indicate a big difference in tooth-by-tooth dental health in Finland. A huge decline in caries attack was seen from the 1960 cohort to the 1980 cohort, but a deterioration of dental health in premolars and second molars is clearly seen in the 1980 cohort in the 1990s. PMID- 12635774 TI - Periodontal treatment needs in a follow-up study among adults with diabetes in Finland. AB - We assessed changes in periodontal treatment needs among patients with diabetes and the risk factors involved in this phenomenon. The sample consisted of 120 dentate subjects, all of whom were regular patients at the Salo Regional Hospital Diabetes Clinic. They underwent periodontal examination in 1999 and were re examined in 2001. The drop-out rate was 4%. Clinical periodontal examination included identification of visible plaque, the presence of calculus, and use of the Community Periodontal Index of Treatment Needs (CPITN) index. Diabetes related factors consisted of information about duration of diabetes, complications, and HbA1c values. Oral health-related factors were collected by questionnaire. The CPITN index proved to be insensitive to change. Pathological pockets (CPITN 3 or 4) were found in 80% of subjects (n = 115) and 48% of sextants (n = 627); in 1999, the corresponding rates were 77% and 49%. The tooth based individual CPITN index (code 3 or 4) revealed periodontal deterioration in 38 patients. Smoking and infrequent interdental cleaning were significant factors explaining periodontal deterioration in logistic regression. Risk factors should be taken into account when planning prevention, treatment, and supportive periodontal therapy strategies. In diabetes care, the common risk factor approach can be implemented to promote oral health among individuals with diabetes. PMID- 12635775 TI - Oral treatment need and oral treatment intention in a population enrolled in long term care in nursing homes and home care. AB - The aim of the study was to evaluate the realistic oral treatment need in a population in southern Sweden enrolled in long-term care (LTC), in nursing homes (NH), or home care (HC), taking into consideration treatment intention. Every third individual enrolled in LTC was selected after proportionally stratifying a total of 866 subjects according to gender. Of these, 732 (85%) were available for a simple clinical oral health evaluation in their own homes. Dental status, oral mucosal status, oral hygiene status, oral mucosal inflammation, and oral mucosal friction were assessed by observational examinations; suspected malignancies were also noted. Oral treatment need was expressed in accordance with the Treatment Need Index (TNI) as no, minor, major, or urgent, while treatment intention was expressed in accordance with the Treatment Intention Index (TII) as the aim to relieve, delay, maintain, or improve. The rationale for using the TII is to offer subjects in this generally frail population oral treatment at an appropriate level, taking their medical condition into consideration. It was found that 61% of the sample had a need not just for an oral health evaluation but also for additional dental treatment, 31% to be accomplished by prophylactic and 30% by reparative or emergency measures; only 1% were estimated to be in urgent need. Furthermore, one manifest and one suspected oral malignancy were found. The results indicate that realistic oral treatment need, guided by the examiner's estimation of the appropriate treatment intention, is modest in this population, but that regular oral screening is mandatory. PMID- 12635776 TI - Epidemiological dental indices and self-perceived oral health in adolescents: ecological aspects. AB - The aim of this study was to investigate the correlations between epidemiological indices (objective) and self-perceived oral health (subjective) in adolescents at school level, and to study gender differences in epidemiological indices and in self-perceived oral health. The study comprised two sets of data from Skaraborg County, Sweden: 1. Self-reported questionnaires answered by adolescents at all senior level schools (n = 9,559, 13-15 years). 2. Epidemiological indices based on clinical registrations of oral health in 13-15-year-old adolescents were collected in all 17 municipalities (n = 7,899). Simple and partial Pearson correlation coefficients were used to study correlations between subjective and objective oral health in the adolescents at school level. Gender differences in adolescents' subjective and objective oral health were estimated using a logistic regression model. The correlations between epidemiological index registrations and self-perceived oral health were weak. The strongest correlations were found between epidemiological indices and self-perceived gingival bleeding: 0.416 between the DS (decayed surfaces) index and self-perceived bleeding. Girls were less likely to be satisfied with the appearance of their teeth than boys were in municipalities with clinical good oral health OR 0.76 (95%) CI 0.59-0.98) and with poor clinical oral health OR 0.74 (CI 0.57-0.94). No gender differences were found in the epidemiological index registrations. The currently used epidemiological indices did not reflect adolescents' own perceptions of their oral health at school level and they could not recognize or identify gender differences. Surveillance of oral health in young people should include information on self-perceived oral health. PMID- 12635777 TI - Dental erosion in Icelandic teenagers in relation to dietary and lifestyle factors. AB - High consumption of fruit juices and carbonated drinks has been related to dental erosion. Teenage male Icelanders consume about 800 ml of carbonated drinks per day on average and this corresponds with the main age group and gender of patients seen with erosion. This study examined the prevalence of dental erosion in 15-year-old children in Reykjavik and looked at the association between erosion and some lifestyle factors in a case-control study drawn from the same sample. A 20% sample of the 15-year-cohort population (n = 278) was selected. Dental erosion was classified by location and severity (1 = enamel erosion; 2 = dentine erosion; 3 = severe dentine erosion) and was seen in 21.6% of subjects (68.3% male; 72% scored as grade 1; 23% grade 2; 5% grade 3). Control subjects were the first healthy subjects examined after a case subject had beens diagnosed. In the case-control study, information was gathered by multiple-choice questionnaire on symptoms of gastric reflux, tooth sensitivity, some lifestyle and dietar-factors. Several lifestyle and dietary factors, previously shown to be significantly related to dental caries in Icelandic teenagers, showed no significant relationship to tooth erosion. Although dietary factors are probably important in causing erosion, dietary recall questionnaires did not help in discovering individuals likely to develop erosion. PMID- 12635778 TI - Does caries prevention correspond to caries status and orthodontic care in 0- to 18-year-olds in the free public dental service? AB - The aim of the study was to evaluate the coverage of and time used for caries preventive actions by patient and by dentist's characteristics in relation to caries status and orthodontic treatments in patients from birth to 18 in public dental care in Helsinki City Health Department, Finland. Data on caries prevention for the age groups 0- to 18-year-olds in 1999 were analyzed in relation to patient age, caries status, orthodontic care, and dentist's characteristics. Our study comprised a total of 48,040 patients treated by 140 dentists. Main outcome measures were the coverage of and time used for repeated caries prevention performed during visits following the check-up visit. Nine percent of those under age 6 received repeated caries prevention, 29% of the 6- to 15-vears-olds and 24% of the 16- to 18-year-olds. Nineteen percent in the no caries group (DMFT/dmft = 0) and 36% in the high-caries group (DMFT/dmft > 0 and DT + dt > or = 3) received repeated caries prevention. The average time a dentist used in repeated caries prevention was 3.6 min (s 3.1: median 2.8; range 0.2-15.4 min) for those in the no-caries and 8.1 (s 6.2; median 7.3; range 0.0-31.8 min) for those in the high-caries group. In each category of caries status, the average time used for repeated caries prevention was shorter for those not under orthodontic treatment. In conclusion, the variation in dentists' caries preventive treatment practices is wide showing both over- and underuse of caries preventive actions as judged by needs of patients. PMID- 12635779 TI - Changes in plaque fluoride levels by school-based fluoride rinsing and tablet programs in Bangladesh. AB - The aim of this study was to evaluate the effects of school-based fluoride rinsing and tablet programs on plaque fluoride levels. A total of 42 children (8 9 years) were selected from two neighboring schools in Dhaka, Bangladesh. After caries recordings, vestibular plaque samples from the maxillary central incisors and mandibular first molars were collected and frozen on day 0 (baseline). The subjects of one school (rinsing group, n = 15) rinsed with 0.05% NaF, while those from the other school (tablet group, n = 16) used a 0.5 mg F tablet on the 5 school days during a 3-week period. The surfaces were sampled on the first and last school day every week. The fluoride and protein contents of each sample were analyzed using micro-techniques. The median plaque fluoride levels were 9.1 ppm at baseline in the rinsing group and 2.5 ppm in the tablet group (P < 0.05). This difference could in part be related to reported fluoride toothpaste usage. After 4 days on the fluoride programs, plaque fluoride levels in the rinsing group increased to 27.3, 24.5 and 14.2 ppm in the 3 consecutive weeks. The corresponding values after tablet usage were 8.0, 6.5 and 7.1 ppm, respectively. After 3 days without fluoride during the weekends, levels declined towards baseline values in both groups. Hence, the plaque fluoride levels in 8 to 9-year old Bangladeshi children were increased by both rinsing and tablet programs, but the effect was not detectable 3 days later. PMID- 12635780 TI - A four-year longitudinal study of palatal plate therapy in children with Down syndrome: effects on oral motor function, articulation and communication preferences. AB - The orofacial function in 20 children with Down syndrome was evaluated after 4 years of palatal plate therapy in 9 of the children (PPG); the remaining 11 were untreated age-matched controls (CG). All 20 children had received continuous orofacial physical therapy from their speech therapist during the treatment period. A clinical extra- and intraoral examination was performed, including oral motor function, facial expression, the occurrence of malocclusions, and hypertrophic tonsils. A questionnaire requesting data on breathing patterns, drooling, eating problems, and communicative preferences was answered by the parents. An articulation assessment was performed by two speech and language pathologists blinded to the treatment status of the children in order to find out whether the palatal plate had stimulated to improved oral speech behavior. The results for oral motor function showed significant differences between the groups in favor of the PPG for the summary variables for: visible tongue (P < 0.01), visible tongue during non-speech periods (P < 0.05), and lip-rounding during spontaneous speech (P < 0.01). During non-speech time, the PPG had their mouths open significantly less than the CG (P < 0.05). Expressivity of facial expression on a visual analog scale in the PPG scored 75.6 +/- 13.3 compared to 51.8 +/- 25.7 in the CG (P < 0.05). The intraoral examination showed that 6/9 children in the PPG and 7/11 in the CG had enlarged tonsils, resulting in more than 50% inter tonsillary space reduction. Despite these findings, and no significant differences between the groups with respect to mouth/ nose breathing, nocturnal snoring was significantly less in the PPG than in the CG (P < 0.05), according to the parental questionnaire. After 4 years of palatal plate therapy, orofacial function had improved significantly in the 9 PPG children and specifically in terms of tongue position and lip activity. PMID- 12635781 TI - Occurrence of vertical bone defects in dentally aware individuals. AB - A vertical (angular) defect is a radiographic diagnostic phenomenon of severe periodontal destruction. The objective of the present study was to investigate the prevalence and severity of vertical defects in a population of dentally aware individuals at 2 different points in time, 1982 and 1992. Intra-oral radiographs of 251 individuals in 1982 and 247 individuals in 1992 in the age range 21-70 years were assessed with regard to presence or absence of vertical defects adjacent to a maximum of 28 teeth per person. A vertical bone defect was defined as a one-sided bone resorption of the interdental marginal bone > or = 2 mm that had a typical angulation towards either the mesial or distal aspect of the root. 'The prevalence of vertical defects in 1982 was 38% increasing from 11% in age 21 30 years to 64% in age 51-70 years, and 27% in 1992 increasing from 7% to 47% in the same age groups, respectively. The number of defects per person increased with age. The majority of affected individuals had 1-2 defects and only about 5% were severely affected. Vertical defects were more common in the posterior as compared to the anterior region of the dentition and the distribution of defects within the maxilla as well as the mandible typically revealed a right-hand to left-hand side symmetry. Vertical defects seem to be a rare phenomenon in dentally aware individuals. In particular, multiple defects were observed only in a minor proportion. Both prevalence and severity increased with age. PMID- 12635782 TI - Cytotoxicity of liquids and powders of chemically different dental materials evaluated using dimethylthiazol diphenyltetrazolium and neutral red tests. AB - The purpose of this study was to assess and compare the cytotoxicity of liquid and powder components of chemically different dental materials using 2 basic unspecific cell culture methods. Three chemically cured glass ionomers (Fuji II, Lining cement, and Ketac Silver), 1 light-cured glass ionomer (Fuji II LC), and 2 chemically cured acrylates (Swedon and Super Bond) were tested. The liquids were diluted 1:10 in cell culture medium. The liquids from chemically cured acrylates were further diluted 1:100, 1:1000, and 1:10000. Extracts were made by incubating the powders in cell culture medium for 24 h at 37 degrees C according to the ISO standard 10993-12. The cytotoxicity was assessed in transformed mouse fibroblasts (L-929) using two viability assays, dimethylthiazol diphenyltetrazolium (MTT) and neutral red (NR). Severe cytotoxicity was observed when testing powder extracts of Swedon, Fuji II, and Lining cement, whereas powder extracts of Ketac Silver, Fuji LC, and Super Bond induced slight to non-cytotoxicity. All of the 1:10 liquid dilutions were severely cytotoxic in the MTT assay. In the NR assay, however, four 10% dilutions were severely cytotoxic and 4 moderately cytotoxic. Further dilution of the liquids of the chemically cured acrylates reduced the toxicity, while the Super Bond catalyst was severely cytotoxic even as the 1:100 dilutions. PMID- 12635784 TI - Retrospective evaluation of implants--is there a need? PMID- 12635783 TI - Inflammatory mediators and radiographic changes in temporomandibular joints of patients with rheumatoid arthritis. AB - The aim of this study was to investigate the relation between the inflammatory mediators tumor necrosis factor alpha (TNFalpha) and serotonin (5-HT), the inflammatory markers erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP), as well as rheumatoid factor (RF) and thrombocyte particle concentration (TPC) in blood versus temporomandibular joint (TMJ) radiographic changes in patients with clinical TMJ involvement by rheumatoid arthritis (RA). Twenty patients were included. Venous blood was collected for quantification of the mediators, markers, and TPC. The radiographic signs of erosion, flattening, sclerosis, subchondral pseudocyst, and osteophyte as well as radiographic grade were investigated with computed tomography. The median (IQR) plasma levels of TNFalpha and 5-HT were 0 (13) pg/ mL and 13 (22) nmol/L, respectively, while serum level of 5-HT was 1360 874) nmol/L ESR, CRP, and TPC were abnormally high in 53%, 250%, and 15% of the patients, respectively. The most frequent radiographic signs were sclerosis (75%), erosion (50%), and flattening (30%). Erosion was found to be associated with high TPC and flattening with high plasma level of TNFalpha. In conclusion, patients with clinical TMJ involvement by RA show an association between high level of TPC and TNFalpha in plasma versus radiographic signs of joint bone destruction. PMID- 12635785 TI - Arthroscopy for osteoarthrosis of the knee is seldom necessary. PMID- 12635786 TI - Expansion of chondrocytes for tissue engineering in alginate beads enhances chondrocytic phenotype compared to conventional monolayer techniques. AB - Chondrocytes are known to dedifferentiate when cultured in monolayer culture, which may compromise the efficacy of cartilage repair systems in which cells are expanded by repeat passage in monolayer prior to implantation. We tested the hypothesis that repeat passage in alginate beads can provide sufficient expansion of cells, while producing cells with enhanced chondrocytic phenotype. Bovine articular chondrocytes were seeded in 2% alginate beads or in monolayer. 4 passages at 7-day intervals were performed. Values of 9.1 days for monolayer expansion and 12.5 days for alginate expansion were estimated for a 10-fold increase in cell number. For assessment of chondrocytic and fibroblastic phenotype, expanded cells were seeded in alginate beads or on glass coverslips and cultured for 7 days. On subsequent seeding in alginate, cells which had previously been subcultured in alginate showed higher levels of both DNA and GAG synthesis than cells passaged in monolayer. Furthermore, the alginate-passaged cells retained a chondrocytic phenotype, indicated by synthesis of type II collagen and chondroitin-6-sulphate, while cells passaged inmonolayer synthesised type I collagen, indicating a fibroblastic phenotype. In conclusion, expansion of cells for autologous cartilage repair systems, using subculture within alginate beads, provides a potentially attractive alternative to monolayer expansion. PMID- 12635787 TI - Distribution of chondroitin 4-sulfate epitopes (2/B/6) in various zones and compartments of articular cartilage in guinea pig osteoarthrosis. AB - We studied changes in the morphology of tibial articular cartilage in early guinea pig osteoarthrosis (OA) at 6 and 12 months of age with quantitative light microscopy, and the distribution of chondroitin-4-sulfate with quantitative ultrastructural immunolabeling, using the 2/B/6 epitope. Labeling was correlated to previous chromatography findings concerning proteoglycan (PG) concentration in animals of the same age. The cell volume fraction had decreased at 12 months in the superficial zone of cartilage with OA (medial condyle) as well as in cartilage without OA (lateral condyle), being lower medially than laterally. The PG concentration differed between the zones and matrix compartments. Medially, a reduction in PG concentration occurred between 6 and 12 months in the interterritorial compartment of the two uppermost zones. Laterally, the concentrations increased. In general, the pericellular PG concentration was higher than the interterritorial in the two uppermost zones. The striking variation in structural and labeling responses in the various zones and compartments indicate a heterogeneous tissue response in guinea pig OA that will probably affect sampling in biochemical analyses of cartilage homogenates and synovial fluid. PMID- 12635788 TI - Determination of variations in gene expression during fracture healing. AB - The genetic make-up and physiological state of a cell or tissue in an organism interact to determine the level at which specific genes are expressed. Identifying genes differentially expressed between 2 genetic or physiological states often gives insight into the molecular mechanisms controlled by the process in question. Various methods have been devised to identify differentially expressed genes and to quantify the expression of differentially regulated genes at the RNA or protein level. These methods are most accurate when the experimental samples are derived from highly controlled and reproducible sources, such as cultured cells. However, no simple in vitro models have been developed to study all biological processes and some are still best studied in the context of the whole organism. Using bone fracture healing as a model, we quantified the expression of 2 housekeeping and 2 regulatory genes during this complex biological process to determine the statistical parameters required to study differential gene expression in tissue samples derived from entire organisms. Our analysis shows that 5 samples in each group are needed to identify genes differentially expressed by a factor of 3 between 2 physiological or genetic states. PMID- 12635789 TI - Periprosthetic bone loss after cemented total hip arthroplasty: a prospective 5 year dual energy radiographic absorptiometry study of 15 patients. AB - In this prospective 5-year study, we determined the periprosthetic bone loss after cemented total hip arthroplasty (THA) in 15 patients using dual energy X ray absorptiometry (DXA). A reduction in the periprosthetic bone mineral density (BMD) of 5-18% occurred in all Gruen regions, or regions of interest (ROI), during the first 3 months after THA. The bone loss continued up to 6 months in almost all ROIs. From 1 to 5 years, we found only minor changes in BMD in periprosthetic bone. After the follow-up, the mean greatest bone loss (26%) was seen in the femoral calcar area. The reduction in mean BMD was 5% in men, and 16% in women. The lower the preoperative BMD, the higher was the postoperative bone loss. We found that after the phase of acute bone loss, further loss was minimal, reflecting merely the normal ageing of bone after uncomplicated THA. PMID- 12635790 TI - Poor results with the Shuttle Stop: resorbable versus nonresorbable intramedullar cement restrictor in a prospective and randomized study with a 2-year follow-up. AB - In a randomized prospective study we compared 2 femoral plugs as regards probable migration of the restrictor, cement leakage and possible early aseptic loosening. In group I, the femoral canal was plugged with a resorbable Shuttle Stop (HC Implants, BV. Leiden, The Netherlands). In group II, a nonresorbable restrictor of polyethylene was used (De Puy, UK). Both groups were identical as regards age, gender and primary disease. All patients were followed for at least 2 years after surgery. Postoperatively, we found more failures with displacement of the femoral plug and cement leakage in group I than in group II. At the 2-year follow-up, we found no differences in stem loosening or grade of radiolucent lines in the cement-bone interface. We can not recommend the Shuttle Stop for femoral canal sealing in total hip replacememt. PMID- 12635791 TI - Treatment of displaced femoral neck fractures: a randomized minimum 5-year follow up study of screws and bipolar hemiprostheses in 100 patients. AB - In a randomized study of 100 patients having displaced femoral neck fractures (Garden types 3-4), we compared internal fixation using 2 von Bahr screws (n = 53) with a bipolar Variokopf prosthesis (n = 47) . The duration of surgery was shorter and the blood loss was less in the group with screws. Dislocation of the prosthesis occurred in 7/47 patients, all within 4 months. After a minimum follow up of 5 years, 34 of 53 patients had been reoperated on in the screw group and 3/47 patients in the prosthesis group. No differences in patient mortality were noted between the groups after 2 or 5 years. The bipolar prosthesis seems to be a suitable alternative for primary treatment of displaced femoral neck fractures. PMID- 12635792 TI - Factors predisposing to dislocation of the Thompson hemiarthroplasty: 22 dislocations in 338 patients. AB - In a series of 338 patients, we have retrospectively analyzed technical and anatomical factors, which may predispose to a dislocation of the Thompson hemiprosthesis. 22 patients (7%) had at least 1 dislocation during the 6-month follow-up. The most significant independent factor predisposing to dislocation was the use of a posterior approach (dislocation rate 16%). We examined the radiographs and data on operations in the 22 patients, using 79 random patients without dislocation as controls. Factors correlating with an increase in the incidence of dislocation were the length of the residual femoral neck > 0.5 cm in short patients (< 165 cm), and considerable change in the postoperative offset of the hip. Acetabular measurements showed no correlation to the dislocation. Our findings suggest that the main factors predicting dislocation are technical and not related to anatomical measurements. PMID- 12635793 TI - Traumatic dislocation of the hip with separation of the capital epiphysis: 5 adolescent patients with 3-9 years of follow-up. AB - We reviewed 5 cases of type I epiphyseal fracture with dislocation of the femoral head from the acetabulum in adolescent patients. All children had an open reduction and screw fixation. In all cases, the femoral head developed avascular necrosis. The clinical result after a mean of 3-9 years' follow-up was good according to the Merle d'Aubigne-Postel scale. Despite necrosis, 2 heads developed spherically after treatment: one which had a primary physeal resection and fixation, the other after an autogenous bone graft in the screw track following removal of the screw. PMID- 12635794 TI - The proximal femoral nail (PFN)--a minimal invasive treatment of unstable proximal femoral fractures: a prospective study of 55 patients with a follow-up of 15 months. AB - In 1996, the AO/ASIF developed the proximal femoral nail (PFN) as an intramedullary device for the treatment of unstable per-, intra- and subtrochanteric femoral fractures. In a prospective study, we treated 55 patients having proximal femoral fractures with the PFN from 1997 to 2000. In 34 patients, we achieved what was close to anatomic reduction of the main fracture fragments. Immediate full weight bearing was permitted in 49 patients. During the follow-up period of 15 months, complications occurred in 12 patients. 2 patients had a cut out of the implant because we used too short proximal gliding screws. In 5 patients, closed fracture reduction could not be done and open fracture reduction with use of cerclage became necessary. Careful surgical technique and modification of the PFN can reduce the high complication rate. In conclusion, the PFN is a good minimal invasive implant of unstable proximal femoral fractures, if closed reduction is possible. If open reduction of the fracture becomes necessary and several fragments are found (especially of the greater trochanter), we prefer to use a dynamic hip screw (DHS) with the trochanter stablizing plate. PMID- 12635796 TI - Low-load resistance muscular training with moderate restriction of blood flow after anterior cruciate ligament reconstruction. AB - We performed a prospective study to determine the effects of introducing low-load muscular training with moderate restriction of blood flow during the first 16 weeks after reconstruction of the anterior cruciate ligament. 44 subjects (average age 29 (18-52) years) were randomized into a group that trained restriction of blood flow (group R, n = 22) and a group that trained without restriction (group N, n = 22). Both groups followed the same training schedule. Evaluations of knee extensor and flexor torques before surgery and 16 weeks after it showed a significant increase in muscular strength in group R as compared to group N. The preoperative/16-week postoperative ratio of the cross-sectional area of the knee extensor muscles showed a statistically significant enlargement in group R as compared to group N. 16 weeks after surgery, the short diameters of type 1 and type 2 fibers of M. vastus lateralis tended to be larger in group R (n = 8) than in group N (n = 8), although the differences were not significant. These findings show that low-load resistance muscular training during moderate restriction of blood flow is an effective exercise for early muscular training after reconstruction of the anterior cruciate ligament. PMID- 12635795 TI - Osteoporosis in 5 elderly women with pubic osteolysis. AB - 5 elderly women developed pubic osteolysis after spontaneous fracture of the pubic bone. Radiographs showed gradual progression of the osteolysis, followed by callus formation, and bone union after 4-5 months. Bone mineral density was low in all patients. White blood cell count, erythrocyte sedimentation rate, and C reactive protein were normal. Urine deoxypyridinoline was high, but serum osteocalcin normal. Elderly women with spontaneous fractures and osteolysis of the pubic bone should be considered for evaluation of osteoporosis and treatment. PMID- 12635797 TI - Functional treatments for acute ruptures of the lateral ankle ligament: a systematic review. AB - Our aim with this systematic review was to assess the effectiveness of various functional treatments for acute ruptures of the lateral ankle ligament in adults. We performed an electronic database search using MEDLINE, EMBASE, COCHRANE CONTROLLED TRIAL REGISTER and CURRENT CONTENTS. We evaluated randomized clinical trials describing skeletally mature subjects with an acute rupture of the lateral ankle ligament and compared functional treatments for inclusion in this study. 9 trials met our inclusion criteria. Two reviewers independently assessed the quality of these trials and extracted relevant data on treatment outcome. Where appropriate, results of comparable studies were pooled. Individual and pooled statistics are reported as relative risks (RR) for dichotomous outcome and (weighted) mean differences (W)MD) for continuous outcome measures with 95% confidence intervals (95% CI). Heterogeneity between the trials was tested using a standard chi-square test. Persistent swelling at short-term follow-up was less with lace-up ankle support than with semi-rigid ankle support (RR 4.2 95% CI 1.3 14), an elastic bandage (RR 5.5; 95% CI 1.7-18) and tape (RR 4.1; 95% CI 1.2-14). A semi-rigid ankle support required a shorter period for return to work than an elastic bandage (WMD 4.2; 95% CI 2.4-6.1) (p = 0.7). One trial reported better results for subjective instability using the semi-rigid ankle support than the elastic bandage (RR 8.0; 95% CI 1.0-62). Treatment with tape resulted in more complications, mostly skin problems, than that with an elastic bandage (RR 0.1; 95% CI 0.0-0.8). We found no other statistically significant differences. We conclude that an elastic bandage is a less effective functional treatment. Lace up supports seem better, but the data are insufficient as a basis for definite conclusions. PMID- 12635798 TI - Spontaneous rupture of the Achilles tendon is preceded by widespread and bilateral tendon damage and ipsilateral inflammation: a clinical and histopathologic study of 60 patients. AB - 60 consecutive patients with spontaneous rupture of the Achilles tendon (AT) underwent surgery. Biopsies were taken at the operations from the site of the rupture, the proximal part, the calcaneal insertion, and the peritendium of the injured tendon. A percutaneous needle biopsy was taken from the contralateral (uninjured) AT. On histological examination, collagen degeneration, tenocyte necrosis, and acute inflammation were found at the rupture site in all cases. In the proximal part and at the insertion, degeneration was present in 56/56 and 51/55 of the cases, necrosis in 55/56 and 50/55, and acute inflammation in 49/56 and 35/55, respectively. The severity of the histological changes decreased from the rupture site to the proximal part to the site of insertion, and showed no relation to the age of the patients or the time from the rupture to the operation. Peritendineal vascular changes were minor. In the contralateral AT, degeneration and necrosis were present in 47/50 and 42/50 of the cases, respectively, but the severity of the changes was less than in the injured tendon. Acute inflammation was present in only 1 case. Spontaneous rupture of AT seems to be preceded by widespread, bilateral damage of the tendon and widespread ipsilateral acute inflammation. PMID- 12635799 TI - Kinesthetic sense of the shoulder in patients with impingement syndrome. AB - A proprioceptive deficit is an important determinant of disability in various shoulder disorders, such as instability and osteoarthrosis. In 15 patients with impingement syndrome stage II (Neer 1983), who were treated by arthroscopic subacromial decompression, we measured movement sense by determining threshold levels for the perception of motion of the shoulder. The patients were placed in a specially designed chair allowing continuous passive motion of the shoulder joint, while avoiding cutaneous, auditory and visual stimuli. To assess movement detection thresholds, passive abduction movements of the shoulder were performed at a starting angle of 60 degrees, an amplitude of 10 degrees and an angular velocity of 1.3 degrees/s. Before surgery, all patients had higher threshold levels for the perception of motion in their affected shoulders then in the other side. After decompression, proprioception had improved on the decompressed side, but was unchanged on the other side. PMID- 12635800 TI - Correlation between rotator cuff tear and glenohumeral degeneration. AB - We studied the occurrence and correlation between a rotator cuff tear and glenohumeral degeneration. 44 cadaveric shoulders (22 right) were obtained from 32 subjects (18 females), mean age 73 (62-86) years and without a history of systemic diseases. Rotator cuffs were exposed and tear size (14 shoulders) was measured after removal of soft tissue and deltoid. Articular cartilage damage of the glenoid and humeral head was recorded by photography. A grading system of 1 (intact), 2 (mild) and 3 (severe) was used to determine the severity of cartilage damage. The area of articular cartilage damage was calculated using the Sonic Digitizer Analyzing System. The area of articular cartilage damage to the glenoid and the humeral head in the rotator cuff tear group was 32% and 36%, respectively. It was greater than that in the groups without a tear, which was 6% in the glenoid and 7% in the humeral head. However, it was not correlated with the size of the tear. Most of the articular cartilage damage in massive and large rotator cuff tears was located in the anterior-inferior portion of the glenoid and in the posterior portion of the humeral head. The articular cartilage damage area of the glenoid was correlated with that of the humeral head. In conclusion, the area of glenohumeral degeneration was greater in the rotator cuff tear group and was located in a specific site. PMID- 12635801 TI - Recalcitrant posttraumatic nonunion of the humerus: 23 patients reconstructed with vascularized bone graft. AB - We treated 23 patients with recalcitrant posttraumatic humeral shaft nonunion with vascularized bone grafts (fibula 10, femur 10 and scapula 3). 21/23 patients healed primarily. Venous thrombosis in the graft necessitated postoperative thrombectomy in 2 patients. Complications at the donor site were trivial. We used a vascularized fibular graft in patients with a large bone defect and with poor intrinsic stability of the nonunion site and a corticoperiosteal femoral graft in atrophic nonunion without a substantial bone defect. The scapula graft is easy to transfer to the surgical neck of the humerus on its pedicle. PMID- 12635802 TI - Fractures of the distal radius in low-demand elderly patients: closed reduction of no value in 53 of 60 wrists. AB - To determine the value of reduction of fractures of the distal radius in the very elderly and low-demand or demented patient, we assessed 60 fractures in 59 patients for the reduction achieved and maintained. The mean patient age was 82 (65-93) years. All fractures were initially reduced under regional or general anaesthesia. In 44 dorsally displaced fractures, reduction failed in 7 cases initially, and 37 lost reduction during the following weeks of immobilisation in plaster. In 9 wrists with volarly displaced fractures, reduction was achieved in 6; all malunited. A total of 53/60 fractures healed in a malunited position. We found no correlation between fracture classification, initial displacement, and final radiographical outcome. On the basis of these observations we conclude that reduction of fractures of the distal radius is of minimal value in the very old and frail, dependent or demented patient. PMID- 12635803 TI - Fracture of a ceramic acetabular insert after ceramic-on-ceramic THA--a case report. PMID- 12635804 TI - Bilateral recurrent atraumatic dislocation of the hip joints--a case report. PMID- 12635805 TI - Free osteochondral fragment caught in the acetabular fossa in the osteochondritis dissecans after Legg-Calve-Perthes'--disease-report of 2 cases. PMID- 12635806 TI - Injury to the deep femoral artery during proximal locking of a distal femoral nail--a report of 2 cases. PMID- 12635807 TI - Smallpox: a 21st century killer? PMID- 12635808 TI - The development of an Alzheimer's disease channel for the Michigan Interactive Health Kiosk Project. AB - The overall objective of this article is to describe the development and implementation of an Alzheimer's disease (AD) module for the Michigan Interactive Health Kiosk Project. This project provides access to health information via 100 kiosks located in public places throughout the state (e.g., churches, senior centers, schools, shopping malls, grocery stores, hospitals) using interactive multimedia software. Because the kiosk screen was designed to resemble a television set, program modules are referred to as "channels." The AD channel is designed to increase knowledge and awareness of the disease among the general public. After an overview of the overall project, the steps involved in developing the AD channel and strategies designed to monitor its use are described, including touch-screen usage data and an on-line user survey. Finally, marketing and promotion of the channel and the implications of this initiative for disseminating information about AD are discussed. PMID- 12635809 TI - Agenda setting for smoking control in Japan, 1945-1990: influence of the mass media on national health policy making. AB - Agenda setting is regarded as a key process in policymaking. This study first examines the trends in newspaper articles on smoking and health and the debates on the issue in the Diet in Japan for the period 1945-1990. Then relationships of those articles and debates with national administrative actions are analyzed. Although the media helped set the agenda in the Diet before the emergence of the nonsmokers' rights movement, it did not do so thereafter. On the other hand, media reports continued to be associated with various aspects of administrative policy making throughout the study period and played an important role in mobilizing administrative agencies. Effects of mass media on agencies were regarded as largely independent of the debates in the Diet. It is also noted that simple "scientific" reports on the health hazards of smoking had no association either with agency action or with Diet debates. This indicates that issue building, which consists of creating a package of ideas about the facts, the causal theories, the responsibilities, and the feasible solutions, is important when scientific facts are to be dealt with by policymakers. PMID- 12635810 TI - The influence of famous athletes on health beliefs and practices: Mark McGwire, child abuse prevention, and Androstenedione. AB - When Mark McGwire broke Roger Maris's home run record in September of 1998, he was instantly declared an American hero and held up as a positive role model for teenagers and young adults. The extensive media attention focused on McGwire made the general public aware of his use of a muscle-building dietary supplement, Androstenedione. It also increased the public's awareness of McGwire's public service to prevent child abuse. The present research assesses audience involvement with McGwire through parasocial interaction and identification, and the effects of that involvement on audience knowledge of and attitudes toward Androstenedione and child abuse prevention. Results indicate parasocial interaction with an athlete regarded as a public role model likely leads to audience identification with that person, which in turn promotes certain attitudes and beliefs. In this case, parasocial interaction and identification with Mark McGwire was strongly associated with knowledge of Androstenedione, intended use of the supplement, and concern for child abuse. Implications of this research for featuring celebrities in health communication campaigns are discussed. PMID- 12635812 TI - A case against "binge" as the term of choice: convincing college students to personalize messages about dangerous drinking. PMID- 12635811 TI - Political ill-health coverage: professional-ethical questions regarding news reporting of leaders' ailments. AB - While reporting on Presidential health has increased of late, there has been very little discussion of the professional-ethical issues involved from the perspective of the journalist, especially when such medical information is not disclosed voluntarily and/or the public official is someone other than the President. Within the general issue of press freedom vs. the right to privacy, and in light of relevant laws, judicial rulings, legal scholarship, and especially journalistic professional ethics, this essay raises and discusses several questions that reporters should take into account when considering whether to publish unauthorized medical information about public officials: 1) do officials have any right to medical privacy? 2) if so, before, during, and/or after tenure? 3) what is the significance of false vs. non-disclosure by the official? 4) are there limits to informed speculation? 5) what types and degree of ill health justify reporting? 6) regarding what level of elected and/or appointed officials? Other ancillary questions are noted as well: means of newsgathering; obsolescence of news item; extenuating circumstances justifying not publishing such news; low IQ as a health problem; who is to be considered a journalist; appropriate sanctions for going over the line. The article's conclusion explains why the issue of ill health reporting of public officials will become even more problematic in the coming years. PMID- 12635813 TI - Baseline lipophilicity relationships in human cytochromes P450 associated with drug metabolism. AB - From analyses of human P450 substrates and their physicochemical properties, it is apparent that baseline lipophilicity relationships exist for over 70 substrates of eight drug-metabolizing P450 enzymes from families CYP1, CYP2, and CYP3. Equations of the general form shown below result in all cases investigated thus far: deltaG(bind) = adeltaG(part) + b where a is the slope of the line which can be termed the hydrophobicity factor of the enzyme active site, possibly being related to the extent of hydrophobic amino acid residues lining the heme pocket; b is the intercept on the y axis and can be regarded as the sum of nonhydrophobic interactions between enzyme and substrate; deltaG(bind) is the free energy change for substrate binding to P450, based on the relationship deltaG(bind) = RTlnKm where Km is the Michaelis constant, and deltaG(part) is the free energy change for partitioning between n-octanol and water based on the relationship deltaG(part) = -RTlnP where P is the n-octanol/water partition coefficient. These findings facilitate the analysis of P450 enzyme-substrate binding interactions and provide information about the likely hydrophobic character of human P450 active site regions. This shows that there are common interactions for certain numbers of substrates in each case composed of hydrogen bonding and pi-pi stacking, the extent of which varies from one P450 enzyme to another. PMID- 12635814 TI - Cytokine regulation of P-glycoprotein. AB - P-glycoprotein (Pgp) is a membrane bound transporter involved in the disposition of many endogenous compounds and xenobiotics. Alterations in Pgp expression and activity can significantly affect the disposition of Pgp substrates. Infection and inflammatory stimuli have also been shown to alter drug disposition. However, the specific effects of inflammation on Pgp expression and activity are not well understood. This paper evaluates and summarizes the current literature on the effects of cytokines and inflammation on mRNA and protein expression as well as functional activity of Pgp in whole animal models, primary rodent hepatocytes and human carcinoma cell lines. PMID- 12635815 TI - Interactions of herbs with cytochrome P450. AB - A resurgence in the use of medical herbs in the Western world, and the co-use of modern and traditional therapies is becoming more common. Thus there is the potential for both pharmacokinetic and pharmacodynamic herb-drug interactions. For example, systems such as the cytochrome P450 (CYP) may be particularly vulnerable to modulation by the multiple active constituents of herbs, as it is well known that the CYPs are subject to induction and inhibition by exposure to a wide variety of xenobiotics. Using in vitro, in silico, and in vivo approaches, many herbs and natural compounds isolated from herbs have been identified as substrates, inhibitors, and/or inducers of various CYP enzymes. For example, St. John's wort is a potent inducer of CYP3A4, which is mediated by activating the orphan pregnane X receptor. It also contains ingredients that inhibit CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4. Many other common medicinal herbs also exhibited inducing or inhibiting effects on the CYP system, with the latter being competitive, noncompetitive, or mechanism-based. It appears that the regulation of CYPs by herbal products complex, depending on the herb type, their administration dose and route, the target organ and species. Due to the difficulties in identifying the active constituents responsible for the modulation of CYP enzymes, prediction of herb-drug metabolic interactions is difficult. However, herb-CYP interactions may have important clinical and toxicological consequences. For example, induction of CYP3A4 by St. John's wort may partly provide an explanation for the enhanced plasma clearance of a number of drugs, such as cyclosporine and innadivir, which are known substrates of CYP3A4, although other mechanisms including modulation of gastric absorption and drug transporters cannot be ruled out. In contrast, many organosulfur compounds, such as diallyl sulfide from garlic, are potent inhibitors of CYP2E1; this may provide an explanation for garlic's chemoproventive effects, as many mutagens require activation by CYP2E1. Therefore, known or potential herb-CYP interactions exist, and further studies on their clinical and toxicological roles are warranted. Given that increasing numbers of people are exposed to a number of herbal preparations that contain many constituents with potential of CYP modulation, high-throughput screening assays should be developed to explore herb CYP interactions. PMID- 12635816 TI - Arsenic: environmental and health aspects with special reference to groundwater in South Asia. Foreword. PMID- 12635817 TI - The spatial pattern of risk from arsenic poisoning: a Bangladesh case study. AB - Arsenic poisoning in Bangladesh has been one of the biggest environmental health and social disasters of recent times. About seventy million people in Bangladesh are exposed to toxic levels of arsenic (0.05 mg/L) in drinking water. It is ironic that so many tubewells have been installed in recent times to provide drinking water that is safe from water-borne diseases but that the water pumped is contaminated with toxic levels of arsenic. Along with the clinical manifestations, some social problems have also emerged due to arsenic toxicity. Analysing the spatial risk pattern of arsenic in groundwater is the main objective of this paper. Establishing the extent of arsenic exposure to the people will facilitate an understanding of the health effects and estimating the population risk over the area. This paper seeks to explore the spatial pattern of arsenic concentrations in groundwater for analyzing and mapping 'problem regions' or 'risk zones' for composite arsenic hazard information by using GIS-based data processing and spatial analysis along with state-of-the-art decision-making techniques. Quantitative data along with spatial information were employed and analyzed for this paper. PMID- 12635819 TI - Food chain aspects of arsenic contamination in Bangladesh: effects on quality and productivity of rice. AB - The total arsenic content of 150 paddy rice samples collected from Barisal, Comilla, Dinajpur, Kaunia, and Rajshahi districts, and from the BRRI experimental station at Rajshahi city in the boro and aman seasons of 2000 was determined by hydride generation-inductively coupled plasma emission spectroscopy (ICP). Arsenic concentrations varied from 10 to 420 microg/kg at 14% moisture content. Rice yields and grain arsenic concentrations were 1.5 times higher in the boro (winter) than the summer (monsoon) season, consistent with the much greater use of groundwater for irrigation in the boro season. Mean values for the boro and aman season rices were 183 and 117 microg/kg, respectively. The variation in arsenic concentrations in rice was only partially consistent with the pattern of arsenic concentrations in drinking water tube wells. There was no evidence from yield or panicle sterility data of arsenic toxicity to rice. Processing of rice (parboiling and milling) reduced arsenic concentrations in rice by an average of 19% in 21 samples collected from households. Human exposure to arsenic through rice would be equivalent to half of that in water containing 50 microg/kg for 14% of the paddy rice samples at rice and water intake levels of 400 g and 4 L/cap/day, respectively. PMID- 12635818 TI - Arsenic groundwater contamination and sufferings of people in North 24-Parganas, one of the nine arsenic affected districts of West Bengal, India. AB - To understand the magnitude of the arsenic calamity in West Bengal, a detailed study spanning 7 years was made in North 24-Parganas, one of the nine arsenic affected districts. Area and population of North 24-Parganas district are 4093.82 sq. km and 7.3 million, respectively. Fourty eight thousand and thirty water samples were analyzed from hand tubewells of North 24-Parganas in use for drinking, cooking and 29.2% of the tubewells were found to have arsenic above 50 microg/L, the maximum permissible limit of World Health Organization (WHO) and 52.8% have arsenic above 10 microg/L, WHO recommended value of arsenic in drinking water. Out of the 22 blocks of North 24-Parganas, in 20 blocks arsenic has been found above the maximum permissible limit and so far in 16 blocks people have been identified as suffering from arsenical skin lesions. From the generated data, it is estimated that about 2.0 million and 1.0 million people are drinking arsenic contaminated water above 10 microg/L and 50 microg/L level, respectively in North 24-Parganas alone. So far, in our preliminary study 33,000 people have been examined at random from arsenic affected villages in North 24-Parganas and 2274 people have been registered with arsenical skin lesions. Extrapolation of the available data indicates about 0.1 million people may be suffering from arsenical skin lesions from North 24-Parganas alone. A sum of 21,000 hair, nail, and urine samples analyses from arsenic affected villages show 56%, 80%, and 87% people have arsenic in biological specimen more than normal/toxic (hair) level, respectively. Thus, many may be subclinically affected. Due to use of arsenic contaminated groundwater for agricultural irrigation, rice and vegetable are getting arsenic contaminated. Hence there is an additional arsenic burden from food chain. People from arsenic affected villages are also suffering from arsenical neuropathy. A followup study indicates that many of the victims suffering from severe arsenical skin lesions for several years are now suffering from cancer or have already died of cancer. PMID- 12635820 TI - Chemical fate of arsenic and other metals in groundwater of Bangladesh: experimental measurement and chemical equilibrium model. AB - The presence of toxic level of inorganic arsenic in groundwater used for drinking in Bangladesh and neighboring India is unfolding as one of the worst natural disaster in the region. The purpose of this work is to ascertain the chemical fate of arsenic and other metals in groundwater of Bangladesh. A combination of techniques was used to measure 24 metals, 6 anions, Eh, pH, dissolved oxygen, conductivity, and temperature to understand the distribution of components which were then used in computational chemical equilibrium model, MINEQL+, for detailed speciation. It was found that the fate of arsenic and its speciation were inextricably linked to the formation of hydrous ferric oxide (HFO) and its kinetic. The HFO induced natural attenuation removes 50-75% of total arsenic in first 24 h through a first order kinetics. Adsorption on HFO is the predominant mode of removal of arsenic, iron, manganese, and most trace metals. The equilibrium model points to the presence of excess active sites for the removal of arsenic. MINEQL+ shows that significantly higher concentration of HFO forming iron is required to remove arsenic below maximum contamination level (MCL) of 50 microg/L than predicted by stoichiometry. The practical implication of this work is the prediction of water quality based on models. PMID- 12635821 TI - Pattern of excretion of arsenic compounds [arsenite, arsenate, MMA(V), DMA(V)] in urine of children compared to adults from an arsenic exposed area in Bangladesh. AB - Urinary arsenic is generally considered as the most reliable indicator of recent exposure to inorganic arsenic and is used as the main bio-marker of exposure. However, due to the different toxicity of arsenic compounds, speciation of arsenic in urine is generally considered to be more convenient for health risk assessment than measuring total arsenic concentration. Additionally, it can give valuable information about the metabolism of arsenic species within the body. In our study, for exposed group--42 urine samples were collected from Datterhat (South) village of Madaripur district, Bangladesh and an average arsenic concentration in their drinking water was 376 microg/L (range 118 to 620 microg/L). For control group, 27 urine samples were collected from a non-affected district, Badhadamil village of Medinipur district, West Bengal, India, where arsenic concentration in their drinking water is below 3 microg/L. The arsenic species in the urine were separated and quantified by using HPLC-ICP-MS. The sum of inorganic arsenic and its metabolites was also determined by FI-HG-AAS. Results indicate that average total urinary arsenic metabolites in children's urine is higher than adults and total arsenic excretion per kg body weight is also higher for children than adults. For arsenic species between adults and children, it has been observed that inorganic arsenic (In-As) in average is 2.36% and MMA is 6.55% lower for children than adults while DMA is 8.91% (average) higher in children than adults. The efficiency of the methylation process is also assessed by the ratio between urinary concentration of putative product and putative substrate of the arsenic metabolic pathway. Higher values mean higher methylation capacity. Results show the values of the MMA/In-As ratio for adults and children are 0.93 and 0.74 respectively. These results indicate that first reaction of the metabolic pathway is more active in adults than children. But a significant increase in the values of the DMA/MMA ratio in children than adults of exposed group (8.15 vs. 4.11 respectively) indicates 2nd methylation step is more active in children than adults. It has also been shown that the distribution of the values of DMA/MMA ratio to exposed group decrease with increasing age (2nd methylation process). Thus from these results we may infer that children retain less arsenic in their body than adults. This may also explain why children do not show skin lesions compared to adults when both are drinking same contaminated water. PMID- 12635822 TI - Speciation of arsenic metabolite intermediates in human urine by ion-exchange chromatography and flow injection hydride generation atomic absorption spectrometry. AB - Biomethylation is considered as the principal metabolic and detoxification pathway for inorganic arsenic in human. The end products of methylation are less toxic and more readily excreted through urine. Therefore, speciation of metabolites in urine is essential to a better understanding of arsenic metabolism, health effects and detoxification ability of individuals exposed to arsenic through drinking water, food and environmental materials. Speciation of inorganic and methylated arsenic in urine is an analytical challenge and often requires expensive instrumentation. We have applied a relatively inexpensive technique for the separation and analysis of various arsenic species, such as, arsenite, arsenate, monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA) in human urine. The technique is based on ion exchange chromatographic separation followed by flow injection hydride generation atomic absorption spectroscopy (FI HG-AAS). The detection limit varies from 1.0 to 2.0 microg/L for various species. The technique has been successfully applied to speciation of arsenic metabolite intermediates in urine samples collected from patients in Hajiganj, a serious arsenic affected area in Bangladesh. Arsenite (AsIII) was found to be the major component in the urine from these patients. Our findings from patients in Hajiganj, Bangladesh are presented in this paper. The technique permits us to carry out arsenic speciation in urine, essential for toxicological studies and possible nutritional intervention in combating arsenic poisoning in Bangladesh. PMID- 12635823 TI - Effects of arsenic on younger generations. AB - Health risks caused by the chronic exposure to arsenic-contaminated groundwater has been recognized in many Asian and Latin American countries. While the health effects among adults have been well documented, those among the younger generations including children, infants, babies, and fetus have been scarcely reported. In this minireview, possible effects of chronic arsenic exposure on the younger generations are discussed in terms of skin manifestations among the children, exposure among newborns and infants especially from the breast milk, child growth and development, reproductive performance. For each of the topics, our preliminary results obtained in an arsenic-contaminated area in Bangladesh will be described, followed by a brief overview on the recent publications dealing with the topic. Although there exist some human data that suggest possible effects on some of the above-mentioned endpoints, the data as a whole is too premature to conclude whether the arsenic poses serious threat to younger generations or not. Considering the public health importance of toxic effects imposed during the early life stages, it is clear that much more scientific efforts should be made to elucidate the possible effects of arsenic on the younger generations. PMID- 12635824 TI - Chronic arsenic toxicity: clinical features, epidemiology, and treatment: experience in West Bengal. AB - Chronic arsenic toxicity due to drinking arsenic-contaminated water has been one of the worst environmental health hazards affecting eight districts of West Bengal since the early eighties. Detailed clinical examination and investigation of 248 such patients revealed protean clinical manifestations of such toxicity. Over and above hyperpigmentation and keratosis, weakness, anaemia, burning sensation of eyes, solid swelling of legs, liver fibrosis, chronic lung disease, gangrene of toes, neuropathy, and skin cancer are some of the other manifestations. A cross-sectional survey involving 7683 participants of all ages was conducted in an arsenic-affected region between April 1995 and March 1996. Out of a population of 7683 surveyed, 3467 and 4216 people consumed water containing As below and above 0.05 mg/L, respectively. Except pain abdomen the prevalence of all other clinical manifestations tested (e.g., pigmentation, keratosis, hepatomegaly, weakness, nausea, lung disease and neuropathy) were found to be significantly higher in As exposed people (water As > 0.05 mg/L) compared to control population (water As level < 0.05 mg/L). The prevalence of pigmentation and keratosis, hepatomegaly, chronic respiratory disease and weakness rose significantly with increasing arsenic concentrations in drinking water. The respiratory effects were most pronounced in individuals with high arsenic water concentrations who also had skin lesion. Therapy with chelating agent DMSA was not found to be superior to placebo effect. However, therapy with DMPS caused significant improvement of clinical condition of chronic arsenicosis patients as evidenced by significant reduction of total clinical scores from 8.90 +/- 2.84 to 3.27 +/- 1.73; p < 0.0001. Efficacy of specific chelation therapy for patients suffering from chronic As toxicity has further need to be fully substantiated. However, supportive treatment could help in reducing many symptoms of the patients. Treatment in hospital with good nutritious diet has been found to reduce symptom score in a subset of placebo treated patients in West Bengal during the course of DMSA and DMPS trial. People should be advised to stop drinking As contaminated water or exposure to As from any other source. The various clinical manifestations should be treated symptomatically. PMID- 12635825 TI - Neuropathy in arsenic toxicity from groundwater arsenic contamination in West Bengal, India. AB - Large number of people from 9 out of 18 districts of West Bengal, India are endemically exposed to arsenic contaminated groundwater due to drinking of tubewell water containing arsenic level above World Health Organization's maximum permissible limit of 50 microg/L. From our ongoing studies on neurological involvement in patients of arsenicosis from different districts of West Bengal, we report our findings in a total of 451 patients of three districts (Murshidabad, Nadia, and Burdwan), comprising 267 males and 184 females with age ranging from 11 to 79 years. They all had arsenical skin lesions, positive biomarkers and identified source of arsenic contaminated water drinking. Peripheral neuropathy was the predominant neurological complication in these patients affecting 154 (37.3%) of 413 patients of Group 1 and 33 (86.8%) of 38 patients of Group 2. Other possible causes and alternative explanations of neuropathy were excluded. The temporal profile in most of the cases (154 of Group 1) were of chronic affection while the 33 patients of Group 2 developed both neuropathy and dermopathy subacutely. Subacutely affected Group 2 patients had much higher incidence of neuropathy. Paresthesias and pains in the distal parts of extremities were much higher in incidence in Group 2 (73.7% and 23.7% respectively) than in Group 1 (18.4% and 11.1%). Distal limb weakness or atrophy was evident in 7.3% in Group 1 and 10.5% in Group 2. Overall, sensory features were more common than motor features in patients of neuropathy and sensory neuropathy was diagnosed in 30% and 76.3% and sensorimotor in 7.3% and 10.5% respectively in Group 1 and Group 2 subjects. Nerve conduction and electromyographic studies performed in 88 cases revealed dysfunction of sensory nerve in 45% and 27% and of motor nerve in 20% and 16.7% of patients with moderate degree and mild degree of clinical neuropathies respectively. Evoked potential studies performed in 20 patients were largely normal except for two instances each of abnormal visual evoked potential and brainstem auditory evoked potential findings. Prognosis was favorable in mild and early diagnosed cases of neuropathy whereas most of the other more severe and late diagnosed cases showed slow and partial recovery or even deterioration. Outcome in neuropathic patients of arsenicosis and long term toxic neurologic effects yet unexplored and unknown remain as matters of future concern requiring close monitoring. PMID- 12635826 TI - Groundwater arsenic contamination, its health impact and mitigation program in Nepal. AB - About 47% of Nepal's total population is living in Terai region and 90% of them are relying on groundwater as their major source of drinking water. About 200,000 shallow tubewells have been installed by different agencies in 20 Terai districts, serving 11 million people. Recently, arsenic contamination of groundwater has been recognized as a public health problem in Nepal. This has sensitized government, national and international nongovernment organizations working on water quality sector to carry out water quality assessment for arsenic in the affected communities. So far, 15,000 tubewells has been tested where 23% samples exceeded World Health Organization guideline value of 10 microg/L and 5% exceeded "Nepal Interim Arsenic Guideline" of 50 microg/L. It is estimated that around 0.5 million people in Terai are living at risk of arsenic poisoning (>50 microg/L). Some recent studies have reported the prevalence of dermatosis related to arsenicosis from 1.3 to 5.1% and the accumulation of arsenic in biological samples like hair and nail much higher than the acceptable level. Though some steps are being taken by government and private organizations to combat the problem, it has not been able to cover all the affected communities. Nepal still needs more research work on arsenic occurrence and effects and mitigation programs simultaneously. PMID- 12635828 TI - Laboratory case identification of arsenic in Ronpibul village, Thailand (2000 2002). AB - Ronpibul village dwellers in the southern part of Thailand have been exposed to arsenic in the water and the environment over three generations. Over the past decades, clean water supplies, utilization and consumption have been introduced to the area. The villagers still use and select rainwater to other forms of potable water. In 2000, the epidemiological survey by Siripitayakunkit (Siripitayakunkit, U. Survey of Chronic Arsenic Poisoning in Ronpiboon, Nakhon Si Thammarat, Thailand, 2000. Proc. 6th International Conference on the Biochemistry of Trace Elements, Guelph, Canada) showed prevalence rate at 24.7%, by using the skin lesion as selection criteria. In 2000-2002, attempt to initiate the local arsenic patient center, we investigated the population at risk in three villages. The laboratory analyses cover urine arsenic level, urine sugar screening and skin lesion classified by dermatologist. The result showed the prevalence of 5.99% of melanosis and 8.67% of hyperkeratosis, 3.84% of urine sugar > 100 mg/dL and 6.33% urine arsenic > 50 microg/g creatinine. There were low to negligible correlation between arsenic urine with urine sugar (r2 = 0.241) and arsenic urine with skin lesion (r2 = 0.058). PMID- 12635827 TI - Genetic polymorphism in p53 codon 72 and skin cancer in southwestern Taiwan. AB - The Pro/Pro polymorphism of p53 codon 72 has been reported to be related to bladder and lung cancer, but its relationship with skin cancer is unclear. We assessed the hypothesis that there is a relationship between the p53 codon 72, Pro/Pro polymorphism, cumulative arsenic exposure, and the risk of skin cancer in a hospital-based case-control study in southwestern Taiwan. From 1996 to 1999, 93 newly-diagnosed skin cancer patients at the National Cheng-Kung University (NCKU) Hospital and 71 community controls matched on residence were recruited in southwestern Taiwan. The genotype of p53 codon 72 (Arg/Arg, Arg/Pro, or Pro/Pro) was determined for all subjects by polymerase chain reaction-restricted fragment length polymorphism (PCR-RFLP). A questionnaire was administered to each subject for collection of demographic information, personal habits, disease history, diet information, and other relevant questions. The Pro/Pro (homozygous) genotype was more frequent in skin cancer patients (cases, 20%; controls, 12%; P = 0.37). Subjects with the susceptible genotype Pro/Pro and heterozygous (intermediate) genotype Pro/Arg had 2.18 and 0.99 times risk of skin cancer than the wild type Arg/Arg (95% confidence interval, 0.74-4.38; 95% confidence interval, 0.44-2.21), respectively. Compared with subjects with 18.5 < BMI < 23, subjects with BMI > 18.5 had 5.78 times risk of skin cancer (95% confidence interval, 1.06 to 31.36) after adjusting for other risk factors. There was no interaction between BMI and genotype, but the sample size was small. The risk of skin cancer did not significantly vary by tumor cell-type. The risk of skin cancer is increased in individuals with the Pro/Pro genotype. Larger, confirmatory studies are needed to clarify the role of constitutional polymorphisms in p53 and skin cancer risk. PMID- 12635829 TI - Arsenic in Australian environment: an overview. AB - The most common source of elevated As concentrations in the Australian environment are attributable to anthropogenic activities. Mining activities have contributed to the contamination of soil and water primarily in Western Australia and Victoria. However, other anthropogenic activities such as agriculture, forestry and industry have also contaminated soil and water at a localized scale. Currently there are over 1000 As contaminated sites previously used as cattle dips for eradicating cattle ticks. Although As contamination of the environment may be severe enough to limit plant growth there appears to be few other reports identifying the impacts of the contaminants on other organisms, such as fish, mammals and humans. In Australia the impacts of metal/metalloid contamination of the environment are often unnoticed or ignored. However, the impacts of elements such as As may pass unnoticed by the public or regulators due to the perception of the minimal impact a contaminant has on the environment. This paper presents an overview of As in the Australian environment including the sources of As contamination, soil, water and plant As content, and the pathways of exposure. PMID- 12635830 TI - The efficacy of monoisoamyl ester of dimercaptosuccinic acid in chronic experimental arsenic poisoning in mice. AB - The therapeutic efficacy of monoisoamyl meso-2,3-dimercaptosuccinic acid (MiADMSA), a new monoester of 2,3-dimercaptosuccinic acid on arsenic induced oxidative stress in liver and kidneys, alterations in hematopoietic system and depletion of arsenic burden was assessed, in mice. Three different doses of MiADMSA (25, 50 or 100 mg/kg) for five consecutive days were administered in chronically arsenic exposed mice (10 ppm in drinking water for six months). Oral administration of MiADMSA particularly at a dose of 50 mg/kg, produced relatively more pronounced beneficial effects on the inhibited blood delta-aminolevulinic acid dehydratase (ALAD), biochemical variables indicative of hepatic and renal oxidative stress and depletion of arsenic concentration in blood, liver and kidneys, compared with intraperitoneal administration of the drug. The treatment with MiADMSA although, produced essential metals imbalance which could be a restrictive factor for the possible therapeutic use of this compound in chronic arsenic poisoning and thus require further exploration. PMID- 12635831 TI - Diagnosis of arsenicosis. AB - Arsenicosis is chronic subclinical or clinical toxicity due to high level of arsenic in body. Diagnosis of arsenicosis was derived by chronological establishment of facts: (a) arsenic as the cause of malady, (b) drinking water (tubewell water) as the vehicle of arsenic, (c) soil as the source of arsenic, (d) mechanism of leaching of arsenic from soil, and (e) cause of prevalence in particular areas of the country. Arsenicosis has been classified by the author into 4 stages, 7 grades and 20 subgrades. Stage I is pre-clinical or grade 0. While clinical features were not found at this stage, high level of arsenic metabolites was observed in urine. As disease progressed to stable phase of grade 0, high level of arsenic was also found in nails, hair, and skin scales. Stage II or clinical stage is divided into 4 grades, (1) Melanosis, (2) Spotted keratosis in palms/soles, (3) Diffuse keratosis in palms/soles, and (4) Dorsal keratosis. Clinical complications are grouped in stage III and grade 5. Malignancy is considered in stage IV and grade 6. There is a concern of both underdiagnosis and overdiagnosis. Therefore, cases of arsenicosis should be cautiously evaluated. Melanosis was the earliest cutaneous sign of clinical arsenicosis. Mild cases of melanosis could only be revealed by a thorough comparison with normal palms. Similarly mild cases of keratosis might not be visible and could only be revealed by careful palpation of palms and soles. Combination of melanosis and keratosis in adults indicated clinical diagnosis of arsenical dermatosis (ASD) that should be confirmed by showing high arsenic concentration in body tissues e.g., nails, hair, skin scales. Isolated melanosis or keratosis in newborn or children below 2 years almost negated the diagnosis of arsenicosis. Genetic melanosis or keratosis is often present since birth. Isolated melanosis or keratosis in adults should be differentiated from non-arsenical dermatosis and proven by absence of high arsenic level in nails and hair. Non arsenical causes of diffuse melanosis, spotted melanosis or leucomelanosis and localized or generalized keratosis can be clinically differentiated from arsenicosis by absence of pigmentation and keratosis in palms/soles. PMID- 12635832 TI - Antioxidants in detoxification of arsenic-induced oxidative injury in rabbits: preliminary results. AB - To assess the oxidative injuries caused by arsenic toxicity in rabbits and evaluate the detoxifying effects of exogenous antioxidants, we administered arsenic trioxide (3-5 mg/kg/day) in rabbits through a feeding tube for seven days. These rabbits were then treated with a recipe of vitamins, zinc, selenium (VZS) or a plant polyphenol or a placebo for the next seven days. Blood samples were collected from ear vein for spectrophotometric assay of reduced glutathione (GSH), thiobarbituric acid reactive substances (TBARS), and nitrite/nitrate (NOx; index of nitric oxide formation) before arsenic administration, seven days after arsenic administration, and seven days after antioxidant treatment. The total arsenic concentrations in hair and spot urine samples of rabbits before arsenic administration were 0.6 +/- 0.21 microg/g and 34.0 +/- 5.9 microg/L, respectively. Administration of arsenic trioxide significantly increased arsenic concentrations in hair and in urine to 2.8 +/- 0.40 microg/g (p<0.001) and 7372 +/- 1392.0 microg/L (p<0.001), respectively. Arsenic administration to rabbits significantly reduced GSH concentration (post-arsenic, 17.5 +/- 0.81 mg/dL vs. pre-arsenic, 32.0 +/- 0.76 mg/dL, p<0.001), increased TBARS concentration (post arsenic, 8 +/- 1.1 microM vs. pre-arsenic, 5 +/- 0.7 microM, p<0.05), and NOx concentration (post-arsenic, 465 +/- 38.5 microM vs. pre-arsenic, 320 +/- 24.7 microM, p<0.001) as compared to the pre-arsenic levels. There was a negative correlation between TBARS and GSH concentrations (r=-0.464, p<0.01) and between NOx and GSH concentrations (r=-0.381, p<0.05) of intoxicated rabbits. The recovery of the depleted GSH was significantly greater in the polyphenols (77.0 +/- 12.0%) or VZS (67.0 +/- 17.0%) treatment groups compared with the placebo group (36.0 +/- 7.0%). The decrease in NOx level of arsenic-treated rabbits was significantly greater in polyphenols treatment group than the placebo group (60.0 +/- 9.0% vs. 17.0 +/- 6.0%, p<0.001). These results indicate that arsenic induces toxicity in rabbits associated with an increase in lipid peroxidation. Arsenic toxicity increases nitric oxide production in the body. Exogenous antioxidants such as polyphenols and recipe of vitamins, zinc, and selenium are useful for arsenic detoxification. PMID- 12635833 TI - A dugwell program to provide arsenic-safe water in West Bengal, India: preliminary results. AB - In 1982, Dr. K. C. Saha, a dermatologist of Calcutta, West Bengal, identified patients with skin lesions from the district of 24 Parganas, leading him and others to search for a cause. The cause was soon identified to be arsenic in drinking water, but even today, 20 years later, large number of people continue to drink arsenic contaminated water and patients are increasing in number. Project Well is a program chosen for implementation in some villages of North 24 Parganas. Arsenic safe drinking water is provided for adopted villages by constructing shallow, concrete dugwells designed to tap the water of the unconfined aquifer, 20-30 feet below ground level, that contains low levels (< 0.05 mg/L) of arsenic in the target region. The traditional dugwell design is modified by use of tube well hand pumps to withdraw water. The project includes community involvement, programs to increase awareness of the need to drink arsenic safe water, and training in monitoring of dugwell water for arsenic and harmful pathogens. Disinfecting of the water and regulating the water hazard diagram are also included in the training program. The plan is to make the system sustainable at the village level using indigenous labor and materials. PMID- 12635834 TI - Non-cancer effects of chronic arsenicosis in Bangladesh: preliminary results. AB - A total of 136 patients were studied in three Upazillas of Laxmipur, Barisal and Madaripur districts were selected for health awareness campaign program. According to the existing data, these villages were ranked as high exposure. Demographic data of the patients investigated are showing pigmentation was seen in all--136 cases, keratosis was found only in 110 cases, and skin ulcer was detected in 13 cases. The predominant features are indicating respiratory sign and symptoms followed by conjunctival congestion and redness of eyes. Weakness was predominant symptom, and all most all cases reported weakness. There were few smokers (n = 16) and analyses were confined to non-smokers (n = 94). A total of the 94 subjects participated in this study. The sex ratio was 1:1.3 (male vs. female). The relative risk of chronic cough increased with age (RR=2.12, for 26 50 age group; RR=2.30 for age group more than 51) reflecting chronic exposure, but the 95% confidence intervals included unity. The relative risk of chronic bronchitis increased with age (RR=2.68, for 26-50 age group; RR=2.30 for age group more than 51) reflecting chronic exposure, but the 95% confidence intervals also included unity. The findings presented here provide evidence that ingestion of inorganic arsenic in drinking water may results in increases risk of chronic cough and bronchitis. PMID- 12635835 TI - Benefits of palliative surgery for far-advanced gastric cancer. AB - BACKGROUND: The optimum strategy for palliative surgery in gastric cancer patients remains undetermined. METHODS: In total, 525 patients who had undergone palliative surgery between 1994 and 2000 were evaluated in terms of operative mortality, survival, and palliative effect. Patients were grouped according to the UICC's classification of residual tumors (R) after the operation: microscopic residual tumor (R1) (N = 104) and macroscopic residual tumor (R2) (N = 421). Gastric resection was performed in all R1 patients and in 257 of the R2 patients. Non-resection procedures were performed in 164 of the R2 patients, including gastrojejunostomies in 64, gastrostomies in 17, jejunostomies in 60, and laparotomies only in 23. RESULTS: The operative mortality did not significantly differ among R1 distal gastrectomies (4.5%), R2 distal gastrectomies (3.3%), and R1 total gastrectomies (2.9%) (p = 0.919). R2 total gastrectomies showed a particularly higher operative mortality (10.9%) than did the other resection procedures. The survival time and palliative duration were significantly longer in patients after palliative resection than after non-resection operations. Postoperative chemotherapy prolonged the survival time of patients after palliative surgery. CONCLUSION: R1 or R2 distal gastrectomies and R1 total gastrectomies have benefits of survival prolongation and symptomatic palliation. However, the use of a total gastrectomy in R2 patients must be selectively reserved for far-advanced cases, otherwise it should be replaced with less invasive procedures to avoid a high operative mortality rate. Postoperative chemotherapy is useful for prolonging survival time. PMID- 12635836 TI - Bridge-plating osteosynthesis of 20 comminuted subtrochanteric fractures with dynamic hip screw. AB - BACKGROUND: A prospective clinical trial was conducted to evaluate the conjunctive use of an extramedullary device and the bridge-plating technique in the treatment of comminuted subtrochanteric fractures with major extension into the femoral shaft. METHODS: A Winquist criteria was used to classify 3 fracture patterns. The type 3 fractures were excluded from this study because of great extent of the fracture zone for which the bridge-plating technique is not indicated. There were 14 men and 6 women, with a mean age of 49 (range, 17-76) years. A dynamic hip screw (DHS) with a long side plate was chosen as the fixation device because of the small learning curve. RESULTS: The fractures united at a mean of 7.6 (range, 3-15) months postoperatively. Mobility was scored at 9 points in 18 patients and 6 points in 2 patients (Mobility score of Parker & Palmer). Pain was absent in 14, mild in 3, and moderate in 3 patients. Two limbs were shortened by 1 and 1.5 cm, respectively. CONCLUSION: Our results indicate that DHS fixation using the bridge-plating technique leads to union of all comminuted Winquist types 1 and 2 fractures without major complications, and it is a valuable alternative to new intramedullary devices. This procedure offers the significant advantage of being less technically demanding. PMID- 12635837 TI - Premature closure of the physeal plate after treatment of a slipped capital femoral epiphysis. AB - BACKGROUND: Treatment of a slipped capital femoral epiphysis (SCFE) should stabilize the epiphysis and prevent complications. Attempting to obtain physeal closure is still controversial. The purpose of this study was to evaluate the clinical results and complications after treatment of SCFE. METHODS: From 1989 to 2000, 12 patients (14 hips) underwent pinning for treatment of SCFE. For acute and acute-on-chronic slippage, longitudinal traction was attempted for reduction. Patients with chronic slippage received fixation in situ. All patients were available for follow-up for an average of 63 months. RESULTS: Nine of the 14 hips had excellent or good functional results, and 5 had fair results. One hip developed avascular necrosis of the femoral head with a fair result. There was no chondrolysis or osteoarthritis of the joint at the most recent follow-up. Physeal closure in 9 hips occurred at an average of 16 months. The change in articulotrochanteric distance averaged 5.4 mm. CONCLUSION: Although most functional results were not adversely influenced after premature closure of the physeal plate of consideration should be given to the device, such as a dynamic screw, that stabilizes the epiphysis and prevents premature physeal closure in patients who have significant growth potential. To understand the influence of growth disturbance of the proximal femur and the effect of dynamic screw fixation, long-term follow-up is mandatory. PMID- 12635838 TI - Surgical removal of the internal limiting membrane for the treatment of a macular hole. AB - BACKGROUND: To evaluate the efficacy of internal limiting membrane (ILM) peeling in the treatment of a macular hole. METHODS: The ocular evaluation included Snellen visual acuity, a slit-lamp examination, indirect ophthalmoscopy, and contact lens biomicroscopy. The macular holes were confirmed using a Watzke-Allen slit beam test. Surgery consisted of a standard 3-port vitrectomy under local anesthesia. The vitreous was removed, and the macular ILM was peeled by creating a small opening and a tear in the ILM with a bent 22-gauge needle around the inner margin of the vascular arcade. The ILM flap was then grasped with end gripping forceps, and a circular capsulorrhesis maneuver was initiated. Next, gas fluid exchange and internal tamponade with 10% C3F8 were performed, followed by postoperative face-down positioning. RESULTS: Thirty-six eyes in 36 patients with idiopathic macular holes from stages 2 to 4 were included. The average follow-up time was 8.9 months. The holes were completely closed in 33 eyes (92%), and visual acuity was improved in 26 eyes (72%). Ten eyes were pseudophakic, 24 of the 26 phakic eyes had an increased density of the cataract after surgery, which was not detected in 2 cases. One of the patients had vitreous hemorrhage and hyphema; no retinal detachment or retinal tear was found in this study. CONCLUSION: Surgery for macular holes using ILM peeling has a high anatomical and functional success rate. PMID- 12635839 TI - Retrospective analysis of 17 liveborn neonates with hydrops fetalis. AB - BACKGROUND: Hydrops fetalis (HF) is a condition with a high mortality rate. The cause may be due to a variety of underlying diseases. In the majority of cases, death occurs antepartum and intrapartum. For those that are born alive, it is difficult to survive. The purpose of this study was to analyze the clinical manifestations, etiologies and outcomes of liveborn babies with hydrops fetalis. METHODS: From October 1995 through May 2001, 17 liveborn neonates that presented with HF were admitted to our neonate intensive care unit (NICU). We were retrospectively reviewed their records. Clinical data including gestational age (GA) at diagnosis and birth, birth weight, Apgar score, maternal and fetal presentations, laboratory data, etiology and outcome were retrospectively collected and analyzed. RESULTS: The mean GA at diagnosis was 30.5 weeks and the mean GA at birth was 33.8 weeks. The male to female ratio was 8:9. Most cases presented with ascites (12/17) and cardiomegaly (8/17). The most common problem faced by the liveborn HF neonates was cardiovascular anomalies (7/17). Seven of these liveborn HF neonates survived. The overall mortality rate of HF in this review was 59%. In comparison with survival cases, those that died were diagnosed earlier, had lower Apgar scores, had more severe acidosis, and had pericardial effusion. CONCLUSION: Recent advances in prenatal ultrasonographic examinations have made early detection of fetal hydrops possible. The mortality rate of these liveborn hydropic neonates without receiving prenatal therapy was high. More effort in prenatal intervention is needed in order to decrease the mortality rate and improve the outcome of neonates with HF. PMID- 12635840 TI - Primary systemic carnitine deficiency presenting as recurrent Reye-like syndrome and dilated cardiomyopathy. AB - Carnitine deficiency syndrome is a rare and potentially fatal but treatable metabolic disorder. I present a 6-year-old girl with primary systemic carnitine deficiency (SCD) proved by very low plasma carnitine level. Her major clinical features included neonatal metabolic acidosis, epilepsy, recurrent infections, acute encephalopathy, and dilated cardiomyopathy with heart failure before 4 years of age. Other features such as hepatomegaly, hypoglycemia, or hyperammonemia were noted around 5 years of age. Her health improved with resolving cardiomyopathy after the use of L-carnitine (50-100 mg/kg/day). Patients with SCD have high morbidity and mortality. If SCD is suggested as a cause of Reye-like syndrome or dilated cardiomyopathy, L-carnitine therapy should be initiated as a diagnostic test immediately, until the definite diagnosis is confirmed. PMID- 12635841 TI - Treating hyponatremia in an empty sella syndrome patient complicated with possible myelinolysis. AB - Hyponatremia as the presenting manifestation of empty sella syndrome is rare. There is little clinical experience in the management of this problem and its possible therapeutic complications. We herein report on a 44-year-old woman with a past history of massive postpartum hemorrhage who was admitted because of hyponatremia and disturbed consciousness. Initial biochemical data suggested the effects of antidiuretic hormone, but fluid restriction alone offered limited benefit. Later, hormonal levels indicated hypopituitarism. Magnetic resonance imaging and cisternography led to a diagnosis of empty sella. Although glucocorticoid substitution was initiated and the clinical condition initially improved, possible myelinolysis subsequently became a complication. With early recognition and immediate replacement of hypotonic fluid, the patient completely recovered. We report this case to illustrate the fact that glucocorticoid substitution and concurrent fluid restriction can probably lead to myelinolysis in empty sella syndrome patients. We suggest that the serum sodium level should be frequently monitored and that much more attention should be paid to the neurologic signs when substituting glucocorticoids in these patients, even though the increment in the serum sodium level is acceptable. Once possible myelinolysis develops, early recognition is critical, and the immediate replacement of hypotonic fluid is suggested. PMID- 12635842 TI - Huge retroperitoneal germinoma presenting with pathological fracture of the spine. AB - Primary retroperitoneal germ cell tumors are extremely rare neoplasms. The most common presenting features are abdominal pain and palpable abdominal masses. Pathological fractures of the spine presenting as bilateral lower leg weakness are exceptionally rare. We describe a 16-year-old girl who developed progressive paraplegia after a minor falling injury. Radiological study demonstrated a huge retroperitoneal tumor with invasion of the T12 vertebral body and spinal canal. A posterior surgical approach was used to perform laminectomy (T12, L1), removal of the intraspinal tumor and internal fixation with transpedical screws (T10, T11 to L2,3), and posterolateral fusion. Postoperative combination chemotherapy for six cycles with cisplatin (100 mg/m2 per day for 1 day every 3 weeks), bleomycin (15 units intravenously weekly for 18 weeks) and etoposide (100 mg/m2 per day for 3 days every 3 weeks) were given and the tumor responded dramatically. The patient had fully recovered without evidence of sequelae or recurrence at 2 years after operation. To the authors' knowledge, this is the first case in which a huge retroperitoneal germinoma presented as pathological fracture of the spine and spinal cord compression. The effectiveness of the postoperative cisplatin-based chemotherapy against this tumor made major retroperitoneal surgery to remove the main tumor mass unnecessary is also demonstrated. PMID- 12635843 TI - Eyewall resection for a uveal malignant melanoma under local anesthesia. AB - A 44-year-old man was referred to our hospital for ciliary body tumor management. A uveal melanoma in the left eye without systemic metastasis was our impression after a series of examinations. We treated this patient with eyewall resection under local anesthesia. Eyewall resection was previously regarded as a complicated procedure that should be performed under general anesthesia. Our surgery was successful, and there were no complications related to the local anesthesia. We followed up this patient for 3 years, and no recurrence was found. Eyewall resection is a good alternative to enucleation for treating a ciliary body melanoma. Under a premise of survival, eyewall resection can maintain part of the visual function and cosmetics. General anesthesia is not indispensable when a patient's health does not allow its use. PMID- 12635844 TI - Delayed gadolinium enhancement in epidural space of the cervicothoracic spine in a patient with spontaneous intracranial hypotension. AB - Spontaneous intracranial hypotension (SIH) due to a spinal cerebrospinal fluid leak is a rare but increasing cause of postural headache. Its extravasated epidural fluid collection tends to be non-enhanced or mildly enhanced on enhanced magnetic resonance (MR) imaging. The mild enhancement of the epidural fluid in SIH is usually attributed to fenestrated neovascularization provoked by an inflammatory component, such as blood, of the fluid collection. In this report, we present a case of SIH with a prominent delayed enhancement of the spinal epidural fluid collection on MR imaging. Subsequent vertebral angiography revealed that this delayed enhancement was related to contrast extravasation from a torn anterior meningeal branch of the right vertebral artery. Therefore, we suggest that contrast extravasation from a torn meningeal vessel may be a possible cause of the enhancement in the spinal epidural fluid of SIH. PMID- 12635845 TI - Nobel Lecture. Protein synthesis, proteolysis, and cell cycle transitions. AB - The discovery of the role(s) of protein synthesis and degradation in the operation of the cell cycle is described. PMID- 12635846 TI - Nobel Lecture. Cyclin dependent kinases and cell cycle control. AB - The discovery of major regulators of the eukaryotic cell cycle is described. PMID- 12635847 TI - What determines the intracellular ATP concentration. AB - Analysis is made of the mechanisms that control the intracellular ATP level. The balance between energy production and expenditure determines the energy charge of the cell and the ratio of [ATP] to the adenylate pool. The absolute ATP concentration is determined by the adenylate pool, which, in its turn, depends on the balance between the rates of AMP synthesis and degradation. Experimental data are discussed that demonstrate an increase in the adenylate pool in response to activation of energy-consuming processes. A hypothesis is proposed according to which variation in the adenylate pool and absolute ATP concentration affords a cell the possibility of additional control over processes fulfillinguseful work. A mechanism involved in this regulation is described using human erythrocytes as an example. The hypothesis explains why different metabolic pathways (protein and DNA syntheses, polysaccharide synthesis, and lipid synthesis) use different trinucleotides (GTP, UTP, and CTP, respectively) as an energy source. This allows the cell to independently control these metabolic processes by varying the individual nucleotide pools. PMID- 12635848 TI - Identification of the putative mannose 6-phosphate receptor (MPR 46) protein in the invertebrate mollusc. AB - Mannose 6-phosphate receptor (MPR 300) protein was earlier affinity purified on phosphomannan gel from the membrane extracts of whole animal acetone powder of a mollusc, unio, in the presence of EDTA (Udaya Lakshmi, Y., Radha, Y., Hille Rehfeld, A., von Figura, K., and Siva Kumar, N. (1999) Biosci. Rep. 19:403-409). In the present study we demonstrate that the unio also contains the putative mannose 6-phosphate receptor (MPR 46) that can be purified on the same gel in presence of divalent metal ions (10 mM each of calcium, manganese, and magnesium), and in the absence of sodium chloride and at pH 6.5. Chicken and Fish cell MPR 46 proteins were purified under these conditions (Siva Kumar, N., Udaya Lakshmi, Y., Hille-Rehfeld, A., and von Figura, K. (1999) Comp. Biochem. & PhysioL 123B:261-265). The authenticity of the receptor is further confirmed by its ability to react with the MSC1 antibody that is specific for MPR 46 protein. Additional evidence for the presence of MPR 46 in molluscs could be obtained by metabolic labeling of mollusc cells Biomphalaria glabrata (Bg cells) with [35S] methionine and cysteine, and passing the labeled membrane extract on phosphomannan gel (at pH 6.5 and 7.0). On elution with mannose 6-phosphate, followed by immunoprecipitation of the column fractions, we identified the putative MPR 46 protein in the Bg cells. When Bg cell MPR 46 was deglycosylated along with chicken MPR 46 (control) both species yielded a single polypeptide corresponding to molecular mass of 26 kDa, suggesting that both contain the same receptor protein. PMID- 12635849 TI - Cyclic AMP production and insulin releasing activity of synthetic fragment peptides of glucose-dependent insulinotropic polypeptide. AB - Synthetic fragment peptides of glucose-dependent insulinotropic polypeptide (GIP) were evaluated for their ability to elevate cellular cAMP production and stimulate insulin secretion. In GIP receptor transfected CHL cells, GIP(4-42) and GIP(17-30) dose-dependently inhibited GIP-stimulated cAMP production (40 +/- 8%; p<0.01 and 15 +/- 6%; p<0.05, respectively), while GIP(1-16) exerted very weak agonist effects on cAMP production. In the clonal pancreatic beta-cell line, BRIN BD11, GIP(1-16) demonstrated weak insulin releasing activity compared with native GIP. In contrast, GIP(4-42) and GIP (17-30) weakly antagonized the insulin releasing activity of the native peptide (23 +/- 6%; p<0.05 and 11 +/- 3%, respectively). These data demonstrate the critical role of the N-terminus and the involvement of regions of the C-terminal domain in generating full biological potency of GIP. PMID- 12635850 TI - Phagocytosis and phagosome maturation are regulated by calcium in J774 macrophages interacting with unopsonized prey. AB - Phagocytosis by neutrophils, macrophages, and other professional phagocytes requires rapid remodeling of actin. Early phagosomes are surrounded by a rim of F actin that is disassembled during phagosomoal maturation. Breakdown of periphagosomal F-actin and phagolysosome fusion are calcium dependent processes in neutrophils interacting with serum-opsonized prey, but appears to be calcium independent in macrophages interacting with serum- or IgG-opsonized prey. In the present study, we found that calcium was necessary for phagocytosis, breakdown of periphagosomal F-actin, and phagosomal maturation in J774 macrophages interacting with unopsonized prey. We also observed that lipophosphoglycan (LPG) from Leishmania donovani promastigotes required calcium to exert its inhibitory effect on macrophage phagocytosis and periphagosomal F-actin breakdown. We conclude that calcium is essential for phagocytosis, depolymerization of periphagosomal F actin, and phagosomal maturation in J774 macrophages interacting with unopsonized prey, as well as for proper functioning of LPG. PMID- 12635851 TI - Spectroimmunochemistry using colloidal gold bioconjugates. AB - Using surface-enhanced infrared absorption (SEIRA) spectroscopy of dry films of colloidal gold (CG) bioconjugates with protein A, it is shown that certain characteristic bands of the protein (e.g., amide I, amide II and some other vibration modes) are essentially affected by the metal surface. Thus, the method may be used for controlling the quality of such bioconjugates. Moreover, it is demonstrated that the biospecific reaction of protein A attached to CG particles with human immunoglobulin G (IgG) results in further essential changes in SEIRA spectra, providing a means for an easy and rapid IR spectroscopic detection of biospecific immunochemical interactions (i.e., spectroimmunochemistry). The results obtained can form a basis for developing test systems for detecting various biospecific interactions. PMID- 12635852 TI - Iron particle labeling of haematopoietic progenitor cells: an in vitro study. AB - We present a method for labeling bone marrow haematopoietic progenitor cells with iron particles. Labeling was assessed by magnetic resonance imaging and electron microscopy. Labeling with iron particles could allow the following by imaging techniques of haematopoietic cells in physiologic and pathologic conditions such as the engraftment of haematopoietic progenitor cells or the migration of myelomonocytic cells in inflammatory diseases. PMID- 12635853 TI - Familial transmission of Continuous Performance Test behavior: attentional and impulsive response characteristics. AB - Continuous Performance Tests (CPTs) provide information on attentional processing and impulsive behavior. The results of previous research that used self-report measures have provided evidence for familial transmission (through genetic and/or environmental influences) of impulsive characteristics. The authors of the present study examined whether the impulsive behavioral parameters that are measured by the CPT also share familial relationships. The researchers asked 26 healthy parent-adolescent pairs to complete the Immediate and Delayed Memory Tasks (IMT/DMT; D. M. Dougherty, 1999; D. M. Dougherty, D. M. Marsh, & C. W. Mathias, 2002), a modified CPT (B. A. Cornblatt, N. J. Risch, G. Faris, D. Friedman, & L. Erlenmeyer-Kimling, 1988; H. E. Rosvold, A. Mirsky, I. Sarason, E. D. Breansome, Jr., & L. H. Beck, 1956), and the Barratt Impulsiveness Scale (BIS; J. H. Patton, M. S. Stanford, & E. S. Barratt, 1995), a self-report measure. The main findings can be summarized as follows: (a) commission errors (but not correct detections) on the IMT and DMT were correlated between parents and their adolescent children, (b) adolescents emitted a higher proportion of commission errors than did their parents, and (c) self-reported impulsivity (i.e., BIS) was correlated with commission errors for parents, but not for adolescents. The findings of this study support the use of an objective behavioral measure of impulsivity to assess familial relationships of impulsivity. PMID- 12635854 TI - Violence: incidence and frequency of physical and psychological assaults affecting mental health providers in Georgia. AB - Threats to the physical safety and psychological well-being of mental health professionals present serious problems, according to 1,131 licensed mental health professionals in the state of Georgia. The professionals included clinical social workers, marriage and family therapists, professional counselors, psychiatrists, psychologists, and social workers. From a total of 6,431 surveys sent out, 1,131 (18%) were returned. The results revealed 2.696 separate accounts of 14,120 acts of physical or psychological assaults committed by 8,292 perpetrators. The results also showed that 61% of the respondents had been victimized in violent acts of a psychological or physical nature and that 29% had feared for their lives at least once during their professional careers. These data are consistent with other research that indicates that 6 out of 10 professionals will be assaulted during their professional careers. PMID- 12635855 TI - Functional anatomy of the null hypothesis and of tests of it. AB - The author compared simulations of the "true" null hypothesis (zeta) test, in which sigma was known and fixed, with the t test, in which s, an estimate of sigma, was calculated from the sample because the t test was used to emulate the "true" test. The true null hypothesis test bears exclusively on calculating the probability that a sample distance (mean) is larger than a specified value. The results showed that the value of t was sensitive to sampling fluctuations in both distance and standard error. Large values of t reflect small standard errors when n is small. The value of t achieves sensitivity primarily to distance only when the sample sizes are large. One cannot make a definitive statement about the probability or "significance" of a distance solely on the basis of the value of t. PMID- 12635856 TI - Some thoughts regarding the sensitivity of t to error in estimated within-group variance. AB - A. J. Riopelle (2003) has eloquently demonstrated that the null hypothesis assessed by the t test involves not only mean differences but also error in the estimation of the within-group standard deviation, s. He is correct in his conclusion that the precision of the interpretation of a significant t and the null hypothesis tested is complex, particularly when sample sizes are small. In this article, the author expands on Riopelle's thoughts by comparing t with some equivalent or closely related tests that make the reliance of t on the accurate estimation of error perhaps more salient and by providing a simulation that may address more directly the magnitude of the interpretational problem. PMID- 12635857 TI - State-dependent memory effects using caffeine and placebo do not extend to metamemory. AB - The authors examined the impact of caffeine on human memory and predictions of memory (i.e., metamemory). On Day 1, 83 college students drank a sweetened beverage containing either caffeine (4 mg/kg body weight) or a placebo before they studied 40 pairs of words. While the participants studied, they predicted their future memory performance for each word pair. On Day 2, the participants again received caffeine or a placebo before the memory test. The participants who drank the same beverage on both days (either caffeine or a placebo) recalled more word pairs than did those who drank different beverages (caffeine on 1 day and a placebo on the other day). In contrast, memory predictions were more accurate when the beverages did not match on both days. These data provide evidence for state-dependent memory when caffeine is used, but not for state-dependent metamemory. People's memory and their predictions of memory can be influenced in different ways if they drink caffeine before they study or take a test. PMID- 12635859 TI - The role of intentional forgetting in employee selection. AB - The authors examined the effects of forbidden information on the employee selection process. They presented the participants with 4 applicants for the position of cashier-stockperson. One of the applicants (the Target Applicant) provided a mixture of forbidden and job-relevant information; the remaining applicants gave no forbidden information. Some of the participants were told before they reviewed the applicants what types of information were to be considered as forbidden, and they were instructed to disregard any such information in the applications. The remaining participants were not aware of the presence of the forbidden information, nor were they instructed to disregard such information. The participants who were instructed to disregard the forbidden information rated the Target Applicant more favorably than did those who were not aware of its presence. Moreover, those in the disregard condition recalled less of the forbidden information and more of the job-relevant information about the Target Applicant than did those in the no-instruction condition. These data patterns support laboratory-based studies on intentional forgetting. PMID- 12635858 TI - An archival study of eyewitness memory of the Titanic's final plunge. AB - A handful of real-life studies demonstrate that most eyewitnesses accurately recall central details (i.e., the gist of what happened) from traumatic events. The authors evaluated the accuracy of archival eyewitness testimony from survivors of the Titanic disaster who witnessed the ship's final plunge. The results indicate that most eyewitness testimony (15 eyewitnesses of 20) is consistent with forensic evidence that demonstrates that the Titanic was breaking apart while it was still on the ocean's surface. Despite the methodological limitations of archival research, the authors provide evidence from a single occurrence traumatic event (with a large-scale loss of life) that the majority of eyewitnesses accurately recall central details. PMID- 12635860 TI - Proteinase-activated receptor-2 expression on cerebral neurones after radiation damage: immunohistochemical observation in Wistar rats. AB - Radiation damage results in blood-brain barrier damage followed by blood plasma transfer into the neuropil. The transferred liquid contains high amounts of biologically active substances/proteinases including factor Xa and a free pool of serum trypsin, which is not bound to antiproteases (alpha1 AT, alpha2 macroglobulin). The aim of this study was to follow up expression of proteinase activated receptor-2 (PAR-2) in the brains of Wistar rats after single exposure to radiation at 26 Gy (60Co, 23 min, 15 sec). After irradiation, the animals were sacrificed on days 10, 20, 30 and 40. Control rat brains served as negative control. Coronal sections of caudal diencephalons were investigated using histology and immunohistochemistry. Polyclonal goat specified antibody against the NH-end of murine and rat PAR-2. Significant PAR-2 membrane positivity of scattered swollen neurons in deeper cortical layers was found in irradiated animals compared with controls. Although this membrane positivity was noticed in all irradiated animals, the most prominent occurred on day 30. Diffuse cytoplasmic positivity was also demonstrated on shrunken neurons in the cortex and hippocampus. Increased cytoplasmic and polarized membrane positivity was also noticed on the neurons of hypothalamic nuclei The causal relationship between blood-brain barrier damage, PAR-2 activation and neurodegeneration has not yet been verified. However, the present findings indicate that PAR-2 mediates a certain cellular response. It remains to be demonstrated whether this is a response to higher concentrations of factor Xa, a free pool of trypsin or other unknown possible proteinases in brain tissue; whether changes in PAR-2 expression are consequences of direct radiation damage to neuronal cells; whether this reaction is protective; and whether primary PAR-2 activation results in neuronal damage. PMID- 12635862 TI - Long-term ethanol administration enhances age-dependent modulation of redox state in different brain regions in the rat: protection by acetyl carnitine. AB - Chronic alcoholism is a major public health problem and causes multiorgan diseases and toxicity. Although the majority of ethanol ingested is metabolized by the liver, it has intoxicating effects in the brain. Evidence is accumulating that intermediates of oxygen reduction may be associated with the development of alcoholic disease. Several studies have shown the capacity of carnitine and its derivatives to influence ethanol metabolism. We have previously demonstrated that preadministration of L-carnitine to rats receiving ethanol significantly reduced fatty acid ethyl esters in different organs and that the carnitine/acylcarnitine system is crucial for maintaining a functional acetyl-CoA/CoA ratio under conditions in which cellular homeostasis is exposed to the deleterious effects of accumulating organic acids. Ethanol, administered to rats for 20 months, induced significant changes in the status of glutathione, primarily in the brain regions of hippocampus and cerebellum, followed by cortex and striatum, where a decrease in reduced glutathione (GSH) and the GSH/oxidized glutathione ratio was found. The same brain regions showed a significant increase in free radical-induced luminescence and hydroxynonenal (HNE), which were associated with decreased GSH reductase activity. Long-term supplementation with acetyl carnitine significantly reduced GSH depletion, particularly in the brain regions of hippocampus, an effect associated with decreased luminescence and HNE formation. In addition, acetyl carnitine treatment increased GSH reductase and arginase activities. Our results indicate that decreased GSH reductase activities associated with thiol depletion are important factors sustaining a pathogenic role in alcohol-related pathologies. Administration of acetyl carnitine greatly reduces these metabolic abnormalities. This evidence supports the pharmacological potential of acetyl carnitine in the management of alcoholic disturbances. PMID- 12635861 TI - Long-term ethanol administration enhances age-dependent modulation of redox state in brain and peripheral organs of rat: protection by acetyl carnitine. AB - Evidence is accumulating that intermediates of oxygen reduction may be associated with the development of alcoholic disease. Free radical-induced perturbation of the oxidant/antioxidant balance in the cell is widely recognized as the main causative factor of age-related disorders. In the present study we investigated the effects of 20 months of ethanol consumption on the antioxidant defense system in different rat organs compared with normal aging in the absence and presence of treatment with L-acetyl carnitine. We demonstrate that aged rats underwent significant perturbation of the antioxidant defense system, as indicated by depletion of reduced glutathione (GSH) content, increased oxidized GSH, free radical-induced luminescence associated with increased hydroxynonenal content and decreased GSH reductase activity. These modifications, observed particularly in brain and liver compared with other organs, were enhanced by long-term alcohol exposure and, interestingly, were significantly reduced with acetyl carnitine supplements. Our results indicate that decreased GSH reductase activity and thiol depletion are important factors in effecting a pathogenic role for oxidative stress in aging and in all situations in which age-correlated and oxidant-induced changes occur, such as in alcoholism. Administration of acetyl carnitine greatly reduces these metabolic abnormalities. Our findings support its pharmacological potential in the management of alcoholic disturbances. PMID- 12635863 TI - Prevalence of C282Y mutation in patients with rheumatoid arthritis and spondylarthritis. AB - Rheumatoid arthritis is an inflammatory joint and systemic disease believed to be of autoimmune origin. Predisposing factors also include genetic factors, such as the presence of alleles HLA-DRB1 *04, (HLA-DRB1 *0401, *0404, *0405 and *0408) and, in other ethnic groups, of subtypes DRB1 *0101, *0102 and DRB1 *1001. These genetic factors are believed to raise the risk of developing the disease. In rheumatoid arthritis, as in other chronic inflammatory diseases, iron metabolism dysfunction has been observed and attributed to inflammation. In hereditary hemochromatosis, tissue sideropexia is associated with a peculiar form of arthropathy. C282Y is a point mutation involving the replacement of a cysteine with a tyrosine at position 282 of the HFE protein. When found in homozygosis, there is a close association with hereditary hemochromatosis, accounting for one of the causes of iron metabolism dysfunction observed in this disease. The aim of this study was to compare the frequency of C282Y in patients with rheumatoid arthritis with that in patients with different forms of spondylarthritis and to correlate these findings with iron metabolism parameters. In the group of patients with rheumatoid arthritis, 2/24 (8.34%) were found to be positive for the C282Y mutation in the case of heterozygosis compared with 3/24 (12.5%) of patients with spondylarthritis. In patients with the C282Y mutation, ferritin levels were significantly higher than those in controls; conversely, serum iron levels were higher in patients with spondylarthritis. Serum transferrin levels, although slightly higher in rheumatoid arthritis patients, showed no statistically significant differences. PMID- 12635864 TI - Simvastatin and markers of endothelial function in patients undergoing continuous ambulatory peritoneal dialysis. AB - Patients undergoing continuous ambulatory peritoneal dialysis (CAPD) are prone to dyslipidemia and have a high risk of cardiovascular death. The aim of this study was to assess the effects of a 6-month treatment with simvastatin (10 mg at bedtime) on markers of endothelial cell injury in 12 hypercholesterolemic CAPD patients. Cholesterol and low-density lipoprotein cholesterol fell significantly after 1 month of therapy. Simvastatin treatment significantly decreased concentrations of vascular cell adhesion molecule and intracellular adhesion molecule after 3 and 6 months of the therapy, respectively. Thrombomodulin decreased significantly after 6 months of the treatment, whereas von Willebrand's factor, P-selectin and E-selectin remained unaltered during simvastatin therapy. Simvastatin, an effective hypolipemic agent, favorably affects endothelial function and may potentially slow the progression of atherosclerosis and confer protection from thrombotic complications in patients with hypercholesterolemia undergoing CAPD. PMID- 12635865 TI - Effects of a low extracellular magnesium concentration and endotoxin on IL-1beta and TNF-alpha release from, and mRNA levels in, isolated rat alveolar macrophages. AB - The aim of this study was to assess the effects of a low extracellular Mg2+ concentration and endotoxin on interleukin (IL)-1beta and tumour necrosis factor (TNF)-alpha release from, and mRNA levels in, rat alveolar macrophages. A low extracellular Mg2+ concentration enhanced the release of both cytokines, and this release was suppressed by a calcium antagonist, nifedipine. Bacterial lipopolysaccharide (LPS) also enhanced the release of both cytokines, and this enhancement was stronger in a low-Mg2+ medium than in control medium. Furthermore, LPS increased the mRNA levels of both cytokines in alveolar macrophages, and this increase was enhanced in low-Mg2+ medium. These results suggest that a low extracellular Mg2+ concentration and LPS stimulate the release of IL-1beta and TNF-alpha from rat alveolar macrophages by increasing the synthesis of both cytokines and by Ca2+ signaling pathways. PMID- 12635866 TI - Mechanisms underlying the enhanced elevation of IL-1beta and TNF-alpha mRNA levels following endotoxin challenge in rat alveolar macrophages cultured with low-Mg2+ medium. AB - We have previously shown that interleukin (IL)-1beta and tumour necrosis factor (TNF)-alpha mRNA levels in rat alveolar macrophages are increased in by endotoxin (lipopolysaccharide; LPS)- stimulation and further enhanced by culturing with low Mg2+ medium. We have now investigated the mechanisms of underlying this enhancement by using some specific signal transduction inhibitors. The enhanced elevation of both mRNAs levels was suppressed by pretreatment with TMB-8 (which inhibits calcium release from the endoplasmic reticulum) or dexamethasone (which inhibits nuclear factor [NF]-kappaB and activator protein [AP]-1), but not with verapamil or nifedipine (which inhibits calcium channels). The enhancment of IL 1beta, but not TNF-alpha mRNA levels, was suppressed by pretreatment with W-7 (which inhibits calmodulin), whereas the enhancement of TNF-alpha mRNA levels was suppressed by pretreatment with U73122 (which inhibits phospholipase C). Curcumin (an inhibitor of AP-1), suppressed the increases in both mRNAs induced by low Mg2+ medium alone, but had no suppressive effect on the levels of either mRNA after LPS-stimulation in low-Mg2+ medium. Pyrrolidine dithiocarbamate (an inhibitor of NF-kappaB) prevented the elevation of TNF-alpha mRNA levels induced by low-Mg2+ medium without LPS-stimulation, but had no suppressive effect on IL 1beta mRNA levels. From these results, we conclude that the enhanced elevation of IL-1beta and TNF-alpha mRNA levels seen in LPS-stimulated alveolar macrophages in low-Mg2+ medium occurs partly via the same, and partly via different, signaling pathways. PMID- 12635867 TI - Effect of extracellular magnesium on secretagogue-evoked amylase secretion in the isolated rat parotid gland segments. AB - This study investigates the effect of perturbation of extracellular magnesium [Mg2+]o on basal and acetylcholine (ACh), noradrenaline (NA) and phenylephrine (PHE)-evoked amylase secretion from isolated rat parotid gland segments. Both zero (0 mM) and elevated (5 mM and 10 mM) [Mg2+]o can significantly (P < 0.05) inhibit basal and secretagogue-evoked amylase secretion compared to the responses obtained in normal (1.1 mM) [Mg2+]o. The inhibitory effect of zero [Mg2+]o was more pronounced compared 10 mM [Mg2+]o. A concentration of 5 mM, [Mg2+]o was less effective at a secretagogue concentration of 10(-5) M but more pronounced in inhibiting amylase secretion when the concentration of the secretagogue was reduced to 10(-6) M. The results indicate that both hypo and hypermagnesaemia are associated with reduced salivary amylase secretion and both conditions may be associated with 'the dry mouth syndrome'. PMID- 12635868 TI - Effects of high doses of magnesium in drinking water and voluntary wheel running on magnesium and calcium concentrations in rats. AB - The effect of high magnesium intake in addition to supplementation and voluntary wheel activity on magnesium and calcium homeostasis was investigated in rats. Thirty-six 5-week-old male Wistar rats were divided into four groups (n = 9). Groups 2 and 4 received five hundred ppm of elemental magnesium as MgCl2 provided in drinking water. After 1 week of acclimatization and 4 weeks of supplement and/or exercise, the animals were fasted and sacrificed. It appears that magnesium supplementation as well as exercise played some significant role in the homeostatic changes of magnesium and calcium. This could be of great significance in better understanding of mineral homeostasis particularly in sports medicine. PMID- 12635869 TI - The influence of magnesium on the activity of some enzymes (AST, ALT, ALP) and lead content in some tissues. AB - Many authors in different studies have reported the antagonism between Mg and Pb. Our previous results suggested that oral Mg treatment have better effect on investigation biochemical parameters (protoporphyrins, aminolevulinic acid--ALA and d-aminolevulinic dehydratase ALA-D) used in evaluating Pb intoxication, then CaNa2EDTA, chelation agents, currently used in therapy of Pb intoxication. The toxic effect of Pb induced considerably modifies the activity of many other enzymes. In this work we have examined the influence of Mg (as alternative therapy of Pb poisoning) on enzymes activity--biochemical markers for general health conditions--aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) in condition of lead intoxication. Many studies showed disturbances of activity ALT, AST and ALP. The aim of this study was to confirm positive effects of Mg intake in condition of such intoxication at the level on activity of investigated enzymes. The experiment was performed on 45 male Wister rats, divided in three groups. I--control group; II--group treated daily for 30 days with 100 mg Pb, per kg body weight and next 60 without Pb treatment (spontaneous detoxication); III group--the same treatment as II group for the first 30 days, but next 60 days rats were treated orally with 40 mg Mg/kg body weight. Activity of AST and ALT was significant increased in condition of Pb poisoning, but ALP activity was significant reduced. Influence of excessive oral Mg treatment was positive: decrease of AST activity and ALT activity, which was probably in correlation with significant elimination of Pb from liver and increase of ALT enzyme activity at the normal level. PMID- 12635870 TI - Effect of magnesium supplementation containing mineral bishofit (MgCl2 x 6H2O) solution and pyridoxine hydrochloride on erythrocyte magnesium depletion and behaviour of rats after three-month alcoholization. AB - In therapy it is known that the combination of vitamin B6 and magnesium is beneficial in the treatment of several forms of primary magnesium deficiency. The purpose of the present study was to investigate the effect of complex magnesium supplementation containing mineral bishofit solution (MgCl2 x 6H2O) and pyridoxine hydrochloride on behavioural and biochemical parameters of magnesium deficient alcoholic rats. A complex magnesium supplementation containing mineral bishofit solution and pyridoxine hydrochloride led both to restoration of magnesium level, and to some correction of behavioural disturbances of animals during chronic alcoholization. PMID- 12635871 TI - A new approach to evaluate magnesium status: determination of exchangeable Mg pool masses using Mg stable isotope. AB - Magnesium (Mg) status is difficult to evaluate using conventional status markers. Radioactive isotopes and, less commonly, stable isotopes have been used as research tools to assess the size of the exchangeable pools of minerals as a new approach to evaluate mineral status. The present paper is a report concerning the exploration of exchangeable Mg pools characteristics with the use of radioactive and stable isotopes and its validation as a method to evaluate Mg status. Recent data obtained in our laboratory using stable isotope of Mg in animals and in humans are presented. Animal studies demonstrated clearly that the exchangeable Mg pools is an interesting approach to evaluate Mg status, as the size of Mg pool 1, 2 and 3 decreased in proportion to Mg deficiency. However more studies are still necessary to better appreciate the relationship between Mg status and exchangeable Mg pool size in humans. PMID- 12635872 TI - Changes in ionised Mg2+ and Ca2+ in maternal serum during weaning in goats. AB - Ionised levels of serum magnesium (Mg2+) and calcium (Ca2+) were measured in HEPES-buffered serum from lactating goats before and during weaning. During this period, there was a significant rise in serum Ca2+, and a concomitant fall in Mg2+. These opposing changes can be explained by the known enhanced absorption of Ca relative to Mg by the gut during lactation coupled with a Ca-induced suppression of PTH-driven renal Ca and Mg reabsorption. A rise in the serum Ca2+/Mg2+ ratio during weaning suggests that this is a period of potential cardiovascular risk for the mother, and merits closer study. PMID- 12635873 TI - Importance of the ratio between ionized and total Mg in serum or plasma: new data on the regulation of Mg status and practical importance of total Mg concentration in the investigation of Mg imbalance. AB - There is an inverse relationship between Mg balance and the ratio ionized Mg/total Mg in serum or plasma: in Mg excess, the ratio is decreased and in Mg deficiency the ratio is increased. It works as if a subtle homeostasic compensatory reaction modified the proportion of the most biologically active fraction of blood Mg in order to reduce the effects of Mg imbalance. Easy, available and unexpensive, the evaluation of total Mg in plasma or serum appears as a better marker than ionized Mg in Mg imbalance: it should be priviledged as the initial investigation in clinical practice. PMID- 12635874 TI - Magnesium in drinking water and the risk of delivering a child of very low birth weight. AB - A few previous studies have looked at the relationship between pregnancy outcome and magnesium nutritional intake and found that magnesium supplementation which could have beneficial effects on perinatal outcome. We performed a study to examine the relationship between the levels of magnesium in drinking water and the risk of delivering a child of very low birth weight (birthweight less than 1500 g; VLBW). The study population comprised 1,781 women residing in 252 municipalities who had a first parity singleton birth between January 1, 1993 and December 31, 1997, and for whom complete information on maternal age, education, gestational age, birth weight, and sex of the baby were available. Data on magnesium levels were obtained from the Taiwan Water Supply Corporation (TWSC). The results of our study show that there is a significant trend toward a decreased risk of having a child of VLBW with increasing magnesium levels in drinking water. Only the specific reversibility of the symptoms through nutritional magnesium in drinking water supplementation might demonstrate the existence of a pathogenic link. PMID- 12635875 TI - Magnesium and calcium in drinking water and the risk of death from esophageal cancer. AB - The possible association between the risk of esophageal cancer and the levels of calcium and magnesium in drinking water from municipal supplies was investigated in a matched case-control study in Taiwan. All eligible esophageal-cancer deaths (2204 cases) of Taiwan residents from 1991 through 2000 were compared with a sample of deaths from other causes (2204 controls), and the levels of calcium and magnesium in the drinking water of these residents were determined. Data on calcium and magnesium levels in drinking water throughout Taiwan were obtained from the Taiwan Water Supply Corporation (TWSC). The control group consisted of people who died from other causes, and the controls were pair-matched to the cases by gender, year of birth, and year of death. Compared to those with magnesium levels below 7.0 mg/l, the adjusted odd ratios (95 per cent confidence interval (CI)) were 0.75 (0.62-0.91) for the group with water magnesium levels between 7.3 and 14.0 mg/l and 0.71 (0.57-0.88) for the group with magnesium levels of 14.3 mg/l or more. The adjusted odd ratios for the relationship between calcium levels in drinking water and esophageal cancer were not statistically significantly different from 1. The results of the present study show that there is a significant trend toward a decreased risk of esophageal cancer with increasing magnesium level in drinking water. PMID- 12635876 TI - Magnesium in drinking water and the risk of death from liver cancer. AB - The possible association between the risk of liver cancer and the levels of magnesium in drinking water from municipal supplies was investigated in a matched cancer case-control study in Taiwan. All eligible liver-cancer deaths (9166 cases) of Taiwan residents from 1994 through 1998 were compared with deaths from other causes (9166 controls). The levels of magnesium in the drinking water of these residents were also determined. Data on magnesium levels in drinking water throughout Taiwan were obtained from the Taiwan Water Supply Corporation (TWSC). The control group consisted of people who died from other causes, and the controls were pair-matched to the cases by year of birth and death. The adjusted odd ratios for the relationship between magnesium levels in drinking water and liver cancer were not statistically significant. Confounding variables, hepatitis B and C infection for example, are probably most likely in the genesis of liver cancer. PMID- 12635877 TI - Comparison of magnesium and zinc levels in blood in end stage renal disease patients treated by hemodialysis or peritoneal dialysis. AB - The abnormal metabolism of macrominerals and trace metals, a frequently overlooked fact may be one of the factors influencing clinical disorders in chronic dialysis patients. The purpose of this study was to compare to what extent maintenance hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) therapy influences the magnesium (Mg) and zinc (Zn) levels in blood of patients treated by these two methods. Additionally, we examined the influence of rhEPO therapy on Hb concentration in erythrocytes, total protein and albumin levels in plasma, and possible impact of rhEPO on Zn and Mg content in blood. Seventy-five ESRD (25 treated and 15 non treated with rhEPO on HD), and (20 treated and 15 non treated with rhEPO on CAPD) patients participated in this study. Forty-five healthy volunteers (HV) served as a reference group. The mean plasma and erythrocytes Mg concentration (mmol/l) in HD and CAPD patients without and with rhEPO treatment was found to be significantly higher than in HV. In HD and CAPD patients treated by rhEPO the mean erythrocytes Mg levels were significantly lower than in non rhEPO treated groups. The inverse relationship between Hb and Mg concentration in erythrocytes in HD and CAPD patients with rhEPO treatment was observed (r = -0.63, P < 0.05 and r = -0.59, P < 0.01 respectively). The mean plasma Zn levels (micromol/l) in HD and CAPD patients with and without rhEPO have been significantly lower than in HV. In patients treated by rhEPO significant increase of Zn erythrocytes levels was found. We was found positive correlation between plasma and erythrocyte Zn levels and Hb concentration in HD and CAPD patients treated by rhEPO (r = 0.35, P < 0.05 and r = 0.4, P < 0.05 respectively). The Hb concentrations, total protein and albumin levels in HD and CAPD with rhEPO therapy were found significantly higher than in patients without rhEPO therapy, but still lower if compared to HV. Mean levels of iron and TIBC in plasma have been found significantly lower in both studied groups treated or non treated by rhEPO, in comparison to HV. In both studied groups of patients without rhEPO or treated by rhEPO plasma GC, GSA, Cr concentrations were significantly higher from those obtained in HV. No statistical correlations were found between Mg, Zn and total protein, albumin, iron levels and TIBC. During rhEPO therapy increased Hb concentration was connected with higher plasma and erythrocytes Zn levels and lower erythrocytes Mg levels. rhEPO may improve the Mg and Zn disturbed metabolism in uraemia. PMID- 12635878 TI - Effect of albumin on neonatal serum ionized magnesium in vitro. AB - BACKGROUND: Human serum albumin (HAS) is used to treat hypoproteinaemia in neonates and as a volume expander. The aim of this study was to quantify the decrease in serum concentration of ionized magnesium ([Mg2+]) when human serum albumin is added to neonatal serum in vitro. METHODS: Human serum albumin was added to 20 cord serum samples of term infants to reach incremental concentrations of 0 to 20.0 g/l and [Mg2+] were measured. RESULTS: Serum [Mg2+] decreased significantly with the addition of serum albumin. At incremental serum albumin concentration of 10 to 20 g/l, which is within the range of the desired aim in the treatment of hypoalbuminaemia, the magnitude of the decrease in serum [Mg2+] was approximately 0.041 to 0.052 mmol/l (10 to 13 per cent) from the average baseline value. CONCLUSION: The addition of albumin causes a decrease in [Mg2+]. From this in vitro study we speculate that fast infusion of albumin in human neonates may potentially cause a clinically significant decrease in serum [Mg2+]. PMID- 12635879 TI - Mechanisms responsible for vasodilation upon magnesium infusion in vivo: clinical evidence. AB - A number of studies have shown that intravenous administration of magnesium (Mg) is an effective treatment for acute coronary syndromes. While mechanisms such as antiarrhythmic, antithrombotic, and myocardial cytoprotective effects may be involved, a vasodilatory effect is among the likely beneficial effects of Mg infusion. The vasodilatory effect of Mg is incompletely understood. On one hand, Mg can act as a physiologic calcium-antagonist, but in addition interest recently has been focused on whether endothelium-derived nitric oxide is involved in Mg induced vasodilation. Another issue in Mg-induced vasodilation is whether the specific Mg salt administered influence the degree of vasodilation. Although Mg sulphate has undergone more clinical studies than Mg chloride, experimental studies suggest that Mg chloride influences vasculature more than Mg sulphate. No clinical research has investigated whether the vasodilatory effect of Mg differs according to the associated anion administered. The present review includes evidence concerning mechanisms of the vasodilatory effect of Mg including endothelium-derived nitric oxide, as well as the possible differences between different Mg salts. PMID- 12635880 TI - Intravenous magnesium sulphate effect on maternal serum and amniotic fluid cytokines levels in preterm labour patients. AB - Our objective was to evaluate the effect of intravenous magnesium sulphate administration to patients with preterm labour on maternal serum and amniotic fluid IL-1beta, IL-6, IL-10 and TNFalpha concentrations. Thirty-six patients at 24-34 weeks of singleton gestation, who presented with contractions (> or = 8 in 60 min) had amniocentesis to rule out intrauterine infection. The patients received intravenous MgSO4 for tocolysis. Twenty-six patients had amniocentesis performed before initiation of MgSO4 (controls) while 10 others had the procedure during tocolytic therapy (study patients). Magnesium, IL-1beta, IL-6, TNFalpha and IL-10 concentrations were measured. Study and control groups were statistically compared using Student t test. Mean magnesium levels were significantly higher in the study group (P < 0.01). There were no significant differences between the cytokines levels in maternal serum and in amniotic fluid between the groups. Our results suggest that the mechanism of magnesium as a tocolytic agent may not be mediated via the examined cytokines. PMID- 12635881 TI - Relationship between hypermagnesaemia in preterm labour and adverse health outcomes in babies. AB - The Magnesium and Neurologic Endpoints Trial (the so-called MagNET Trial) was a randomized clinical trial that was undertaken to establish whether the antenatal usage of magnesium sulphate could protect neonates from having adverse neurologic outcomes. Unfortunately, the trial was suspended after 15 months of enrolment because of excess total paediatric mortality among those exposed to magnesium sulphate. Following our original report and contrary to the original hypotheses, additional analyses of our data have actually shown a statistically significant increase in the risk of neonatal intraventricular hemorrhage, as well as total adverse paediatric outcomes, among those with higher levels of ionized magnesium at delivery. Nonetheless, it has been postulated, but not established, that anions of magnesium other than sulphate could have a more benign, or even beneficial, effect on health outcomes in the neonate. PMID- 12635882 TI - Chronopathological forms of magnesium depletion with hypofunction or with hyperfunction of the biological clock. AB - The main mechanisms of the chronopathological forms of magnesium depletion associate a low Mg intake with various dysregulating biorhythms. The differentiation between forms with hyperfunction and forms with hypofunction of the biological clock is seminal and the main marker is the production of melatonin (MT). The clinical forms of the various patterns of the chronopathological forms of Mg depletion may be central or peripheral. The clinical forms with hyperfunction of the biological clock (marker: increase in MT) may associate diverse expressions of nervous hypoexcitability: depression (i.e. Seasonal affective disease); cephalalgias nocturnal, without photophobia (i.e. cluster headaches); dyssomnia LASPS (advanced sleep phase syndrome) particularly]; asthenia and myalgias (i.e. fibromyalgia, chronic fatigue syndrome). The main comorbidity is found with depressive states. The therapy relies on classical bright light phototherapy, sometimes associated with psychoanaleptics. The clinical forms with hypofunction of biological clock (marker: decrease in MT) may associate various signs of nervous hyperexcitability (HEN): anxiety (from generalized anxiety to panic attacks); cephalalgias diurnal with photophobia (mainly migraine); dyssomnia [DSPS (delayed sleep phase syndrome) particularly, jet lag, night work disorders, age related insomnia, sometimes with inappropriate behaviour; photogenic epilepsia, generalized or focal; some clinical forms of chronic fatigue syndrome and fibromyalgia. The main comorbidity is between migraine and epilepsia. The treatment relies on the diverse forms of darkness therapy, possibly with the help of some psycholeptics: anxiolytics and anticonvulsants. The indications of chromatotherapy remain to be validated. PMID- 12635883 TI - Magnesium deficit and sudden infant death syndrome (SIDS): SIDS due to magnesium deficiency and SIDS due to various forms of magnesium depletion: possible importance of the chronopathological form. AB - Sudden Infant Death Syndrome (SIDS) might be due to the fetal consequences of a Mg maternal deficiency, which might be prevented by simple atoxic nutritional Mg intake by the mother. Various stresses in the pregnant women or in the infant may transform a simple Mg deficiency into Mg depletion which may not be cured by nutritional Mg supplement, but requires a correction of its causal dysregulation. Beside the well established risk factors in baby care and in the environment, it is important to stress the possible role of a primary hypofunction of the biological clock. This may be treated by darkness therapy: total light deprivation at night for the infant and atoxic nutritional Mg supplement for the pregnant women. The place in the prevention of SIDS of Mg therapy for the infant and of the use of melatonin, L-tryptophan and taurine is now uncertain yet. PMID- 12635884 TI - Modifications of magnesium and copper concentrations in serum and arterial wall of patients with vascular diseases related to ageing, atherosclerosis and aortic aneurysm. AB - Relationship between concentration of Mg and Cu in serum and the arterial wall was studied in patients with atherosclerosis obliterans (AO), aortic aneurysm (AA) and in the control group. The effect of ageing was also evaluated. The results showed increased serum concentration of Cu in the eldest control group (50-59 years) when compared to the younger (20-29 years). Serum concentration of Mg was decreased in AO and AA, and that in the arterial wall was lower in AA, than in controls. Concentration of Cu in serum and the arterial wall was higher in AO than in controls. Lower limb ischaemia results in decreased serum Mg and increased Cu in the critical degree as compared with the moderate degree of ischaemia. The ratio Mg/Cu was found lower both in serum and the arterial wall in AO and AA, as a result of ageing and vascular disease, and it could be better marker of ischaemia than individual element concentrations. PMID- 12635885 TI - Alteration of myocardial mechanics in marginal magnesium deficiency. AB - Magnesium has attracted attention as an essential element with diverse roles in the regulation of cardiac contraction. Chronic suboptimal intake of the element results in hypomagnesaemia. Experimental and clinical studies indicate the possibility of a marginal decrease in myocardial magnesium compared to those with sufficient intake. Reduction in extracellular magnesium affects myocardial excitability and contractility predominantly, by modulation of the levels of other ions that have an influence on cardiac mechanics. Majority of the in vitro experiments in isolated ventricular tissue or myocytes record an inverse relation between Mg concentration and inotropic response, mediated probably by enhanced influx of Ca2+ promoting sarcoplasmic reticular Ca2+ release. Paradoxically myocardial contractility is usually compromised in animals on Mg deficient diet or on perfusion of whole heart with low Mg (< 0.5 mM) buffer. In the whole animal or organ, magnesium deficiency induced coronary vasospasm, defective energy metabolism and excessive free radical generation may be important variables acting in concert or independently to affect myocardial function. Electrical excitability is enhanced in magnesium deficiency, and arrhythmic changes are presumed to be mediated by disturbance in K+ homeostasis. Magnesium deficiency has not received the attention it deserves probably due to absence of clinical symptoms. Magnesium deficiency concomitant with stress may be of clinical significance, leading to arrhythmic, hemodynamic and ischaemic changes in the heart. Chronic magnesium deficiency is accompanied by increased free radical generation. Free radicals are known to influence myocardial excitability and contractility. Physiologic and pathologic stress also promotes free radical generation. The additive action of free radical generation in magnesium deficiency and any form of stress may be one of the reasons for enhanced sensitivity to stress in magnesium deficiency. Clinical and experimental data on the cardiac consequences of marginal magnesium deficiency being limited, a number of factors need experimental validation. For example--the extent of change in total and ionized magnesium in the serum and heart, mechanical response of the myocardium to decrease of total and ionized magnesium in the intra- and extracellular milieu; the extent of free radical generation in magnesium deficiency and the cardiac consequence; and also the additive effect of magnesium deficiency and different forms of stress. PMID- 12635886 TI - Pro-fibrogenic effects of magnesium deficiency in the cardiovascular system. AB - Magnesium deficiency is known to produce cardiovascular injury. A large body of experimental evidence supports the postulation that an immuno-inflammatory reaction and increased oxidative stress may damage the myocardium and vasculature in magnesium deficiency. Reparative/reactive fibrosis in response to the injury has, however, received little attention. Recent evidence from a rodent model of acute magnesium deficiency suggests that humoral factors may activate cardiac fibroblasts by a free radical-mediated mechanism and contribute to cardiac fibrogenesis. A similar mechanism may also promote cellular hyperplasia and increased matrix synthesis in the vasculature. PMID- 12635887 TI - PhRMA and the dermatologist. PMID- 12635888 TI - Botanical briefs: the sunflower--Helianthus species L. PMID- 12635889 TI - Androgenetic alopecia in adolescents. AB - Androgenetic alopecia (AGA), or hereditary hair thinning, is a common and unwelcome cause of hair loss in men and women. AGA also occurs in adolescents, though its prevalence in this younger population is not known. Physical appearance is extremely important to most adolescents, and early onset of hair loss can have a definite negative effect on self-image and self-esteem. Minoxidil topical solution is widely used by adults for hair loss, but its use by adolescents has not been systematically evaluated. This article provides an overview of AGA and presents new information on the prevalence and age at onset of hereditary hair thinning in adolescents. In addition, data are presented on the efficacy and proper use of minoxidil topical solution in adolescent boys and girls. PMID- 12635890 TI - What is your diagnosis? Pseudocyst of the auricle. PMID- 12635891 TI - Basal cell carcinoma of the umbilicus: a case report and literature review. AB - We report the case of a 43-year-old woman with basal cell carcinoma (BCC) of the umbilicus. Although BCC is a common skin tumor, only 2 cases of BCC arising within the umbilicus have been reported previously. Our review of the literature shows that truncal BCCs frequently develop in younger patients, often grow larger, and are associated with an increased risk for developing multiple nonmelanoma skin cancers. Therefore, we advocate a low threshold for performing biopsies on umbilical lesions that are atypical in appearance, course, or response to therapy. PMID- 12635892 TI - Vitamin B12 deficiency: a case report of ongoing cutaneous hyperpigmentation. AB - We describe an interesting case of a man with recurrent cutaneous and hematologic manifestations of vitamin B12 deficiency. In this deficiency, the skin, central nervous system, blood, and blood-forming tissues are commonly involved. We describe an overview of vitamin B12 deficiency and the successful treatment of a patient's ongoing cutaneous hyperpigmentation. PMID- 12635893 TI - Defining skin of color. PMID- 12635894 TI - Defining skin of color. PMID- 12635895 TI - Merkel cell tumor presenting as a painful patch lesion on the right arm. AB - Primary small cell cutaneous neuroendocrine carcinoma (Merkel cell carcinoma) is an uncommon, highly malignant, primary cutaneous neuroendocrine carcinoma. Clinically it is seen as a 0.5- to 5.0-cm pinkish purple papule or nodule, usually not ulcerated, on the head, neck, or, less frequently, the roots of the limbs. We present the case of a woman with an atypical clinical presentation of a Merkel cell tumor. PMID- 12635896 TI - A patient with extensive stem cell factor-induced hyperpigmentation. AB - Stem cell factor (SCF) is a cytokine that stimulates development of erythroid precursors and, consequently, may have potential importance in the treatment of certain anemias. We report a case of a young woman with Diamond-Blackfan anemia who received SCF treatment. One effect of SCF treatment is cutaneous hyperpigmentation at the injection site. In contrast to previously reported cases of patients who had fewer SCF injections, her hyperpigmentation was extensive and of major cosmetic concern. SCF may play a role in a number of disease processes. This case demonstrates the need to consider the potential pigmentary side effects of SCF therapy and to make careful selection of the injection sites. PMID- 12635897 TI - Oral cephalosporin antibiotics: an overview of clinical pharmacology and dermatologic applications. PMID- 12635898 TI - Tacrolimus ointment 0.1% produces repigmentation in patients with vitiligo: results of a prospective patient series. AB - The cause of the selective melanocyte destruction in vitiligo may be due to an autoimmune disorder. A series of 15 patients with vitiligo were treated with a topical immunomodulator, tacrolimus ointment 0.1%, twice daily for a minimum of 45 days. Thirteen patients (87%) experienced at least partial repigmentation, and 3 of those patients had greater than 75% repigmentation. Patients with the greatest treatment response likely benefited from concomitant natural sunlight exposure. Further studies investigating the safety and efficacy of tacrolimus ointment either as monotherapy or in combination with other therapeutic measures are warranted. PMID- 12635899 TI - Interval estimation for Weibull-distributed life data under Type II progressive censoring with random removals. AB - This paper explores the problem of interval estimation for parameters of Weibull distributed data, which are Type II progressively censored with random removals. Seven different confidence interval-estimation procedures are considered. Four of them are based on a parametric bootstrapping approach. Others are based on the asymptotic normality method and the likelihood ratio statistic. We conduct a Monte Carlo simulation to evaluate the performance of these procedures based on their lengths and their coverage probabilities. Furthermore, an example is presented to illustrate the application of these procedures. PMID- 12635900 TI - Dunnett's many-to-one test and least square means. AB - Dunnett's many-to-one test is used frequently today, especially in dose-finding studies. Using Dunnett's test, the Type I error level for the comparison between the raw mean of the control and the raw means of the study drug groups can be exactly calculated for the normal data. However, this computability depends on the independence of the raw means. Unfortunately, this independence does not exist for the model-based likelihood estimates (least square means) in the cases of ANCOVA and two-way ANOVA models without interaction for unbalanced data. This paper investigates this dependence between the least square means and derives some new procedures to calculate the joint distribution of the statistic for Dunnett's test. PMID- 12635901 TI - Statistical issues on objective, design, and analysis of noninferiority active controlled clinical trial. AB - In practice, "noninferiority" active-controlled trials have been designed for three different objectives: establishing evidence of efficacy over placebo, preserving a specific percentage of the effect size of the active control, or demonstrating the test treatment is "not much inferior" to the active control. All three objectives can be represented by the same set of statistical hypotheses with the parameters defined differently. The various designs and statistical analysis procedures for active-controlled trials proposed in the literature can be group into two basic types: the historical-controlled trial approach and the cross-study comparison approach. These approaches require some unverifiable constancy assumptions. Under the constancy assumptions, the cross-study comparison uses the estimate effect of active-control treatment as the unbiased estimate of the active/placebo difference in the current noninferiority trial. A normalized Z-statistic is used to test the hypotheses. On the other hand, the historical controlled trial approach uses a conservative confidence limit as if it were a constant to replace the active/placebo difference in the current trial. The two approaches may lead to consistent conclusions only when the constancy assumptions can be supported by a large number of historical studies giving a consistent active-control treatment effect over placebo and that the active control effect does not change over time. PMID- 12635902 TI - Examining outlying subjects and outlying records in bioequivalence trials. AB - The problem of detecting outliers in bioequivalence trials is considered. We formulate the problem as a hypothesis-testing problem under a mean-shift model and propose a test procedure based on the likelihood function. The test statistic has two components: one is to detect whether a specific pharmacokinetic measurement of a subject for certain formulation/drug product is an outlying value; the other is to test whether a subject as a whole is an outlying subject (with unusual high or low bioavailability for all formulations/drug products). Under normality assumption, the proposed procedure is most powerful. The small sample distribution of the proposed test statistic is derived. A numerical example illustrates the use of the procedure. The proposed test is then compared in a simulation study against the Hotelling T2 test, recommended by Liu and Weng (1991) for the use of outlier detection in bioequivalence studies. The results from the simulation study show that the proposed test is more powerful than the Hotelling T2 test. PMID- 12635903 TI - Normalization methods for analysis of microarray gene-expression data. AB - This paper investigates subset normalization to adjust for location biases (e.g., splotches) combined with global normalization for intensity biases (e.g., saturation). A data set from a toxicogenomic experiment using the same control and the same treated sample hybridized to six different microarrays is used to contrast the different normalization methods. Simple t-tests were used to compare two samples for dye effects and for treatment effects. The numbers of genes that reproducibly showed significant p-values for the unnormalized data and normalized data from different methods were evaluated for assessment of different normalization methods. The one-sample t-statistic of the ratio of red to green samples was used to test for dye effects using only control data. For treatment effects, in addition to the one-sample t-test of the ratio of the treated to control samples, the two-sample t-test for testing the difference between treated and control samples was also used to compare the two approaches. The method that combines a subset approach (median or lowess fit) for location adjustment with a global lowess fit for intensity adjustment appears to perform well. PMID- 12635904 TI - Evaluating the agreement of two quantitative assays with repeated measurements. AB - A common task in assay validation is to show the agreement between an assay under investigation and a reference assay. Hence, in the hypothesis setup, we should choose nonagreement as the null hypothesis so that when the null hypothesis is rejected at 5% level of significance, we have a 95% statistical assurance to claim the agreement between two assays. In this paper, we propose a statistical test with nonagreement as the null hypothesis. The calculation of sample size is also given. Some simulation results are provided for illustration. PMID- 12635905 TI - Phase I (or phase II) dose-ranging clinical trials: proposal of a two-stage Bayesian design. AB - We propose a new design for phase I (or phase II) dose-ranging clinical trials aiming at determining a dose of an experimental treatment to satisfy safety (respectively efficacy) requirements, at treating a sufficiently large number of patients to estimate the toxicity (respectively failure) probability of the dose level with a given reliability, and at stopping the trial early if it is likely that no dose is safe (respectively efficacious). A two-stage design was derived from the Continual Reassessment Method (CRM), with implementation of Bayesian criteria to generate stopping rules. A simulation study was conducted to compare the operating characteristics of the proposed two-stage design to those reached by the traditional CRM. Finally, two applications to real data sets are provided. PMID- 12635906 TI - Testing for interaction in studies of noninferiority. AB - We consider the role of interaction tests in the context of active-controlled clinical trials that aim to demonstrate the noninferiority of an experimental treatment compared to a standard (control) treatment. When the subjects can be grouped into strata (e.g., study sites, gender, race, etc.), there may be a desire to determine whether the experimental treatment is noninferior to the standard in each of the strata. We present five possible analysis strategies to test for heterogeneity of relative treatment effects among strata. These strategies are either identical to or straightforward modifications of strategies that can be used to test for interaction when the objective of the study is to show differences rather than noninferiority. The various analysis strategies implicitly depend on different definitions of interaction. Power of the various tests will be low, a phenomenon that often occurs when testing for interaction. We present simulation results to quantify the power and type I error rates under different scenarios and an example to demonstrate the proposed tests. None of the analysis strategies is best under every parameter configuration. The tests may be best used in a descriptive or exploratory manner. Extensions to two-sided equivalence testing are also discussed. PMID- 12635908 TI - Statistical method for analysis of the disease curve in animals with experimental autoimmune encephalomyelitis. AB - Experimental autoimmune encephalomyelitis (EAE) is a procedure used in the laboratory to examine drugs that may have utility in treating multiple sclerosis (MS). The problem of modeling the disease curve in animals with EAE is studied. The classification of animals after each experiment is considered and the chi square test is proposed to test a homogeneity between treatment groups. A mixture type of nonlinear mixed-effects model with repeated measurements is considered, assuming that the onset and/or remission of disease is the fixed effect as well as the random effect. Statistical inference on the parameters of the disease curves is discussed. The proposed model is shown to be efficient for comparing the disease curves with different treatments. Examples concerning the study of the effects of test compounds in EAE are presented to illustrate the proposed model and statistical methodologies. PMID- 12635907 TI - Uising WinBUGS to fit nonlinear mixed models with an application to pharmacokinetic modelling of insulin response to glucose challenge in sheep exposed antenatally to glucocorticoids. AB - Many chronic diseases of adulthood, such as hypertension and diabetes, are now believed to have at least some of their origins before birth. Extensive studies in animal models have identified antenatal exposure to excess glucocorticoids as a leading candidate for the physiological cause of fetal compromise. The resulting adverse intra-uterine environment appears to "program" the individual for higher risk of subsequent disease. We present an analysis of blood glucose and insulin concentrations collected during glucose tolerance tests at 6 and 12 months postnatal age in a cohort of sheep that were treated antenatally with injections of betamethasone (a synthetic glucocorticoid) which, when injected into the mother, cross the placenta to the fetus. A simple pharmacokinetic model, essentially a modification of the single compartment model with first-order absorption and elimination, is developed to describe the time course of glucose concentration and the associated insulin response. The resulting nonlinear mixed model is implemented in a Bayesian framework using the Markov chain Monte Carlo technique Gibbs Sampling via the software package BUGS. This sampling process allows inferences to be made directly about derived quantities with an immediate physical interpretation, such as the maximum insulin concentration in response to glucose challenge. At 6 months postnatal age, sheep treated with antenatal injections of synthetic glucocorticoids had raised insulin concentration in comparison to controls after bolus administration of glucose. This effect persisted to 12 months postnatal age only in the sheep that received multiple doses of glucocorticoids. Moreover, the raised insulin concentration in sheep that received direct injections of synthetic glucocorticoid as fetuses is accompanied by better glucose clearance than in those sheep that received only saline injections, a phenomenon that is not observed in the animals that received maternal injections. It is argued that the fitting of an appropriate statistical model to complex physiological data does not necessarily proclude a result that has a clear interpretation for clinical scientists. PMID- 12635909 TI - Affect as information in persuasion: a model of affect identification and discounting. AB - Three studies examined the implications of a model of affect as information in persuasion. According to this model, extraneous affect may have an influence when message recipients exert moderate amounts of thought, because they identify their affective reactions as potential criteria but fail to discount them as irrelevant. However, message recipients may not use affect as information when they deem affect irrelevant or when they do not identify their affective reactions at all. Consistent with this curvilinear prediction, recipients of a message that either favored or opposed comprehensive exams used affect as a basis for attitudes in situations that elicited moderate thought. Affect, however, had no influence on attitudes in conditions that elicited either large or small amounts of thought. PMID- 12635910 TI - When stereotypes get in the way: stereotypes obstruct stereotype-inconsistent trait inferences. AB - There is a growing body of evidence indicating that people spontaneously make trait inferences while observing the behavior of others. The present article reports a series of 5 experiments that examined the influence of stereotypes on the spontaneous inference of traits. Results consistently showed weaker spontaneous trait inferences for stereotype-inconsistent behavioral information than for stereotype-consistent and stereotype-neutral information. Taken together, the current results suggest that specific spontaneous trait inferences become obstructed by inhibitory processes when behavior is inconsistent with an already activated stereotype. These findings are discussed in relation to stereotype maintenance processes and recent models of attribution in social judgment. PMID- 12635911 TI - Creeping dispositionism: the temporal dynamics of behavior prediction. AB - Four studies tested the hypothesis that temporal distance increases the weight of global dispositions in predicting and explaining future behavior. Study 1 found that the correspondence bias was manifested more strongly in predictions of distant future behavior than of near future behavior. Study 2 found that participants predicted higher cross-situational consistency in distant future behavior than in near future behavior. Study 3 found that participants sought information about others' more global dispositions for predicting distant future than near future behavior. Finally, Study 4 found that participants made more global causal attributions for distant future outcomes than for near future outcomes. The results were interpreted as supporting the assumption of construal level theory that perceivers use more abstract representations (higher level construals) to predict and explain more distant future behaviors. PMID- 12635912 TI - Moral value transfer from regulatory fit: what feels right is right and what feels wrong is wrong. AB - People experience regulatory fit (E. T. Higgins, 2000) when the strategic manner of their goal pursuit suits their regulatory orientation, and this regulatory fit feels right. Fit violation feels wrong. Four studies tested the proposal that experiences of fit can transfer to moral evaluations. The authors examined transfer of feeling wrong from fit violation by having participants in a promotion or prevention focus recall transgressions of commission or omission (Studies 1 and 2). Both studies found that when the type of transgression was a fit violation, participants expressed more guilt. Studies 3 and 4 examined transfer of feeling right from regulatory fit. Participants evaluated conflict resolutions (Study 3) and public policies (Study 4) as more right when the means pursued had fit. PMID- 12635913 TI - Hiding in the crowd: can mortality salience promote affiliation with others who oppose one's worldviews? AB - The present research highlights affiliation defenses in the psychological confrontation with death. In 3 experiments, it was found that mortality salience led to increased affiliation strivings, as indicated by a greater preference for sitting within a group as opposed to sitting alone. Mortality salience actually led to increased affiliation with a worldview-threatening group (Experiments 1 2), even when affiliation with the group forced participants to attack their own worldviews (Experiment 3). Taken together, the findings support a distinct role of affiliation defenses against existential concerns. Moreover, affiliation defenses seem powerful enough to override worldview validation defenses, even when the worldviews in question are personally relevant and highly accessible. PMID- 12635914 TI - Reexamining adaptation and the set point model of happiness: reactions to changes in marital status. AB - According to adaptation theory, individuals react to events but quickly adapt back to baseline levels of subjective well-being. To test this idea, the authors used data from a 15-year longitudinal study of over 24.000 individuals to examine the effects of marital transitions on life satisfaction. On average, individuals reacted to events and then adapted back toward baseline levels. However, there were substantial individual differences in this tendency. Individuals who initially reacted strongly were still far from baseline years later, and many people exhibited trajectories that were in the opposite direction to that predicted by adaptation theory. Thus, marital transitions can be associated with long-lasting changes in satisfaction, but these changes can be overlooked when only average trends are examined. PMID- 12635915 TI - Forgiveness, forbearance, and time: the temporal unfolding of transgression related interpersonal motivations. AB - The investigators proposed that transgression-related interpersonal motivations result from 3 psychological parameters: forbearance (abstinence from avoidance and revenge motivations, and maintenance of benevolence), trend forgiveness (reductions in avoidance and revenge, and increases in benevolence), and temporary forgiveness (transient reductions in avoidance and revenge, and transient increases in benevolence). In 2 studies, the investigators examined this 3-parameter model. Initial ratings of transgression severity and empathy were directly related to forbearance but not trend forgiveness. Initial responsibility attributions were inversely related to forbearance but directly related to trend forgiveness. When people experienced high empathy and low responsibility attributions, they also tended to experience temporary forgiveness. The distinctiveness of each of these 3 parameters underscores the importance of studying forgiveness temporally. PMID- 12635916 TI - Power moves: complementarity in dominant and submissive nonverbal behavior. AB - Two studies examine complementarity (vs. mimicry) of dominant and submissive nonverbal behaviors. In the first study, participants interacted with a confederate who displayed either dominance (through postural expansion) or submission (through postural constriction). On average, participants exposed to a dominant confederate decreased their postural stance, whereas participants exposed to a submissive confederate increased their stance. Further, participants with complementing responses (dominance in response to submission and submission in response to dominance) liked their partner more and were more comfortable than those who mimicked. In the second study, complementarity and mimicry were manipulated, and complementarity resulted in more liking and comfort than mimicry. The findings speak to the likelihood of hierarchical differentiation. PMID- 12635917 TI - Cultural variations on optimistic and pessimistic bias for self versus a sibling: is there evidence for self-enhancement in the west and for self-criticism in the east when the referent group is specified? AB - A culturally relevant framework was used to examine variations on optimistic and pessimistic bias in Westerners and Easterners. Study 1 showed that 136 European Americans compared with 159 Japanese were more likely to predict typical positive events to occur to self than to a sibling. The opposite pattern emerged in the prediction of typical negative events. Study 2 replicated these findings on the basis of predictions for atypical events in 175 European Americans and 130 Japanese. Across both studies, within-groups analyses indicated that European Americans held an optimistic bias in the prediction of positive and negative events, whereas Japanese held a pessimistic bias for negative events. These findings are taken to offer support for presumed cultural differences in self enhancement and self-criticism between Westerners and Easterners, respectively. PMID- 12635918 TI - Work experiences and personality development in young adulthood. AB - This longitudinal study provides an analysis of the relationship between personality traits and work experiences with a special focus on the relationship between changes in personality and work experiences in young adulthood. Longitudinal analyses uncovered 3 findings. First, measures of personality taken at age 18 predicted both objective and subjective work experiences at age 26. Second, work experiences were related to changes in personality traits from age 18 to 26. Third, the predictive and change relations between personality traits and work experiences were corresponsive: Traits that "selected" people into specific work experiences were the same traits that changed in response to those same work experiences. The relevance of the findings to theories of personality development is discussed. PMID- 12635920 TI - Quantifying construct validity: two simple measures. AB - Construct validity is one of the most central concepts in psychology. Researchers generally establish the construct validity of a measure by correlating it with a number of other measures and arguing from the pattern of correlations that the measure is associated with these variables in theoretically predictable ways. This article presents 2 simple metrics for quantifying construct validity that provide effect size estimates indicating the extent to which the observed pattem of correlations in a convergent-discriminant validity matrix matches the theoretically predicted pattern of correlations. Both measures, based on contrast analysis, provide simple estimates of validity that can be compared across studies, constructs, and measures meta-analytically, and can be implemented without the use of complex statistical procedures that may limit their accessibility. PMID- 12635919 TI - Mood regulation and emotional intelligence: individual differences. AB - In 3 studies (Ns = 250, 83, 236), an examination of differences in how individuals experience their emotions (meta-emotion traits of clarity, attention, and intensity) led to the identification of 4 distinct types (overwhelmed, hot, cerebral, and cool). When mood was manipulated, the types differed in how they initially reacted to the emotional situation, how they regulated their mood, and how they made judgments. In particular, one type of individual (the hot type) was more reactive to emotional situations than the others. Another type of individual (the overwhelmed type) regulated mood differently than the others, which led these individuals to make judgments that were also different. Overwhelmed individuals appeared unable or unwilling to avail themselves of critical affective information. PMID- 12635921 TI - Status and solidarity in social comparison: agentic and communal values and vertical and horizontal directions. AB - Social comparison involves positioning the self relative to others on a vertical or status dimension (ranging from upward to downward comparisons) and a horizontal or solidarity dimension (ranging from contrastive to connective comparisons). Across 3 studies in which 389 undergraduates recorded everyday social comparisons (n = 4,417), downward and connective comparisons were rated as more helpful and mood enhancing than upward and contrastive comparisons. The effects of horizontal comparisons were greater for people for whom solidarity was an important value; however, the effects of vertical comparisons were not greater for people who valued status. The roles of the comparison target, topic, and situation were also explored; for example, noticing undesirable features of the target enhanced status but undermined solidarity. PMID- 12635922 TI - Resource loss, resource gain, and emotional outcomes among inner city women. AB - The authors examined a dynamic conceptualization of stress by investigating how economic stress, measured in terms of material loss, alters women's personal and social resources and how these changed resources impact anger and depressive mood. Resource change in women's mastery and social support over 9 months was significantly associated with changes in depressive mood and anger among 714 inner city women. Greater loss of mastery and social support was associated with increased depressive mood and anger. Loss of mastery and social support also mediated the impact of material loss on depressive mood and anger. Resource loss and worsening economic circumstances had more negative impact than resource gain and improving economic circumstances had positive impact, suggesting the greater saliency of loss than gain. PMID- 12635923 TI - Number of older brothers and sexual orientation: new tests and the attraction/behavior distinction in two national probability samples. AB - The extent to which number of older brothers or "fraternal birth order" predicted the 2 main components that researchers have traditionally used to conceptualize sexual orientation-that is, psychological attraction and sexual behavior-was examined in 2 recent national probability samples. In both studies, fraternal birth order predicted same-sex attraction in men, with each additional older brother increasing the odds of homosexual attraction by an average of 38%. Results also indicated that the fraternal birth order/same-sex attraction relationship in men was independent of sexual behavior, including early same-sex behavior. No sibling characteristics predicted sexual orientation in women. Results suggest experience-based theories (e.g., early same-sex play) of the fraternal birth order effect in men are unlikely to be correct. PMID- 12635924 TI - Cost-containment by early, rapid and precise detection of etiologic agents using modern molecular diagnostics? PMID- 12635926 TI - Staphylococcal and streptococcal superantigens: molecular, biological and clinical aspects. AB - Superantigens (SAgs) include a class of certain bacterial and viral proteins exhibiting highly potent lymphocyte-transforming (mitogenic) activity towards human and or other mammalian T lymphocytes. Unlike conventional antigens, SAgs bind to certain regions of major histocompatibility complex (MHC) class II molecules of antigen-presenting cells (APCs) outside the classical antigen binding groove and concomitantly bind in their native form to T cells at specific motifs of the variable region of the beta chain (Vbeta) of the T cell receptor (TcR). This interaction triggers the activation (proliferation) of the targeted T lymphocytes and leads to the in vivo or in vitro release of high amounts of various cytokines and other effectors by immune cells. Each SAg interacts specifically with a characteristic set of Vbeta motifs. The review summarizes our current knowledge on S. aureus and S. pyogenes superantigen proteins. The repertoire of the staphylococcal and streptococcal SAgs comprises 24 and 8 proteins, respectively. The staphylococcal SAgs include (i) the classical enterotoxins A, B, C (and antigenic variants), D, E, and the recently discovered enterotoxins G to Q, (ii) toxic shock syndrome toxin-1, (iii) exfoliatins A and B. The streptococcal SAgs include the classical pyrogenic exotoxins A and C and the newly identified pyrogenic toxins, G, H, I, J, SMEZ, and SSA. The structural and genomic aspects of these toxins and their molecular relatedness are described as well as the available 3-D crystal structure of some of them and that of certain of their complexes with MHC class II molecules and the TcR, respectively. The pathophysiological properties and clinical disorders related to these SAgs are reviewed. PMID- 12635925 TI - Zentralblatt fur Bakteriologie--100 years ago Aladar aujeszky detects a 'new' disease--or: it was the cow and not the sow. PMID- 12635927 TI - Novel vaccination strategies based on recombinant Mycobacterium bovis BCG. AB - In this manuscript, we will review the utilization of Mycobacterium bovis Bacille Calmette-Guerin (BCG) as a vaccine against tuberculosis (TB) and as a carrier system for heterologous antigens. BCG is one of the most widely used vaccines. Novel techniques in genome manipulation allow the construction of virulence attenuated recombinant (r)-BCG strains that can be employed as homologous vaccines, or as heterologous antigen delivery systems, for priming pathogen specific immunity against infectious diseases, including TB. Several approaches are available for heterologous antigen expression and compartmentalization in BCG and recent findings show the potential to modulate and direct the immune responses induced by r-BCG strains as desired. Recent achievements in complete genome analysis of various target pathogens, combined with a better understanding of protective pathogen-specific immune responses, form the basis for the rational design of a new generation of recombinant mycobacterial vaccines against a multitude of infectious diseases. PMID- 12635928 TI - Virulence and the heat shock response. AB - The major adaptive response to elevation in temperature is the heat shock response that involves the induction of many proteins--called heat shock proteins. These include chaperones, proteases, alternative sigma factors and other regulatory and structural proteins. The heat shock response is also turned on by other stress conditions, such as oxidative stress or pH changes. Bacterial entry into the host organism involves a significant environmental change, which is expected to induce the heat shock response. Indeed, some of the heat shock proteins are themselves virulence factors while others affect pathogenesis indirectly, by increasing bacterial resistance to host defenses or regulating virulence genes. The cross talk between heat shock and virulence genes is discussed. PMID- 12635929 TI - Dissemination of pheU- and pheV-located genomic islands among enteropathogenic (EPEC) and enterohemorrhagic (EHEC) E. coli and their possible role in the horizontal transfer of the locus of enterocyte effacement (LEE). AB - We have recently shown that the locus of enterocyte effacement (LEE) of the bovine enterohemorrhagic E. coli RW1374 (O103:H2) resides within a large pathogenicity island (PAI), integrated in the vicinity of the phenylalanine tRNA gene pheV. Here we describe an additional, but LEE-negative genomic island in RW1374 in the vicinity of another phenylalanine tRNA gene, pheU, the sequence of which is identical to pheV. These two genomic islands revealed identity of the left, but a relative variability of their right end sequences. To investigate the mechanism of LEE-PAI distribution in E. coli, we analysed similar junctions in the pheU/pheV loci of additional EPEC and EHEC strains the LEE location of which had not been determined before. By hybridisation of NotI restriction fragments with probes specific for LEE, pheV locus, and pheU locus, the LEE was found linked to either one of these two loci. The results agreed well with recently published phylogenetic data and indicate that in the clones of diarrheagenic E. coli (Dec) Dec 11 and Dec 12, forming the phylogenetic cluster EPEC 2, and in the strains of the most typical serotypes of the Dec 8, belonging to the phylogenetic cluster EHEC 2, the LEE was linked with pheV and not with the pheU locus as previously assumed. Sequence comparison with other pheU- and pheV-located genomic islands from different E. coli pathotypes (uropathogenic E. coli, septicemic E. coli) as well as from Shigella indicated the same structural features at the junctions. These conserved structures suggested a common DNA cassette, serving as common vehicle for horizontal gene transfer of various PAls. In addition, the elements suggest an origin from a common pheU-located ancestor and integration into the chromosome through site-specific recombination. Our results indicate that pheU/pheV-located genomic islands played an important role in the evolution of several PAls in E. coli and related pathogens. PMID- 12635930 TI - Subtyping of pathogenic Escherichia coli strains using flagellar (H)-antigens: serotyping versus fliC polymorphisms. AB - Serotyping of O- and H-antigens is regarded as the gold standard in classification of E. coli for taxonomic and epidemiological purposes similar to the Kaufmann-White scheme for Salmonella enterica. Molecular methods to replace or to support the serotyping have been applied recently. Using the molecular polymorphism of the flagella (H-antigen) gene fliC, more than 220 E. coli strains derived from the E. coli reference collection for O- and H-antigens (The International Escherichia and Klebsiella Centre (WHO)) and from clinical origins have been characterised and a reproducible and clear cut classification with very good correlation to serotyping was found. Only some of the H-antigens have revealed multiple fliC classes and vice versa only rarely some of the fliC classes belong to various H-antigen groups. Since also H-antigen-negative and H antigen non-typeable strains subjected to fliC classification could be typed properly, it is recommended here to use this rapid approach to classify E. coli under routine conditions rather than using classical serotyping. However, scrotyping--in particular using hyperimmune rabbit sera--will remain the gold standard and the task of Reference Centres only, e.g. for defining novel H antigen types. PMID- 12635931 TI - Prevalence of the new, SPI1-like, pathogenicity island ETT2 among Escherichia coli. AB - The new pathogenicity island ETT2 has been identified by PCR and gene probes among various intestinal serovars and pathovars of E. coli, in particular among EHEC/STEC. However, ETT2 was not detected among extra-intestinal and non pathogenic E. coli strains or other enteric bacteria including various S. enterica serovars. A considerable molecular diversity of ETT2 among various E. coli serovars was found. The occurrence of ETT2 among E. coli is independent of the presence of other virulence properties, e.g. the pathogenicity islands LEE, LPA, or HPI. PMID- 12635932 TI - Sequencing of 16S rDNA of Klebsiella: taxonomic relations within the genus and to other Enterobacteriaceae. AB - The 16S rDNAs of 20 strains of Klebsiella were sequenced and used for construction of a phylogenetic tree together with already published Enterobacteriaceae 16S rDNA sequences. The taxonomy within the Klebsiella genus, as reflected by the 16S rDNA tree, was in agreement with existing DNA-DNA hybridisation and numerical taxonomy data, indicating that for Klebsiella, 16S rDNA sequencing is a valid method for identification and taxonomical purposes. Five closely related clusters were found in the Klebsiella genus; Cluster I, K. oxytoca; Cluster II, K. terrigena, Cluster III, K. planticola and K. ornithinolytica; Cluster IV, Enterobacter aerogenes (K. mobilis); and Cluster V, K. pneumoniae. The position of Calymmatobacterium granulomatis within the genus and closest to K. pneumoniae was confirmed. For the species K. oxytoca, data seem to indicate a subdivision into two subspecies. In addition, a biochemically aberrant Klebsiella strain (BEC441) that was included in the analysis could not be assigned to any of the known species, but was found to be closest related to K. oxytoca. Furthermore, the high sequence similarity between the two environmental species K. planticola and K. ornithinolytica does not justify a distinction of the two species. Finally, within a 165-bp stretch of the 16S rDNA sequences, species-specific nucleotides were found. PMID- 12635933 TI - Lack of polymorphism in a Vibrio cholerae O139-specific DNA region encoding the somatic antigen in strains isolated during 1993-1998. AB - Vibrio cholerae O139 Bengal emerged as a second aetiologic agent of cholera in South Asia in late 1992. This new serogroup arose from a Vibrio cholerae O1 strain by deletion of the chromosomal region encoding O1 specificity and acquisition of a novel 35-kb region encoding the O139 specificity. Previous studies indicated significant phenotypic and genotypic changes in O139 isolates over the years since its first appearance. This prompted us to study possible polymorphism in the 35-kb novel region encoding the O139 specificity. A total of 17 V. cholerae O139 isolates originating from different countries and years in South Asia and China, and a single unrelated V. cholerae O139 isolate from Argentina were studied. The 35-kb chromosomal region was amplified as two fragments of 12 and 23 kb in an extended PCR from all isolates. These amplicons were then treated separately with seven different restriction enzymes and separated by agarose gel electrophoresis. The South Asian and Chinese isolates gave identical patterns for the same enzymes, but different patterns for different enzymes, thus exhibiting no polymorphism in the 35-kb region. However, the Argentine isolate gave distinct patterns for most of the enzymes confirming its different origin. This data indicated that the portion of the chromosome encoding the O139 antigen specificity is highly conserved. As found in previous studies, the early O139 isolates were resistant to trimethoprim-sulfamethoxazole (TMP-SMX) and vibriostatic compound, O/129, and CAMP- haemolysin positive. The isolates of later years diverged exhibiting different patterns by pulsed-field gel electrophoresis (PFGE), and becoming susceptible to TMP-SMX and O/129, and CAMP-haemolysin negative. PMID- 12635934 TI - Subtyping of Mycoplasma pneumoniae isolates based on extended genome sequencing and on expression profiles. AB - Mycoplasma pneumoniae isolates from patients, collected over a period of 12 years in Germany, were characterized by various methods (parameters) including multilocus sequence typing, restriction fragment length polymorphisms, Western blotting with mono-specific antibodies directed against selected proteins or with polyspecific antibodies directed against the Triton X-114-soluble protein fraction, and two-dimensional gel electrophoresis. The results for 91 isolates from Germany, which were complemented with 14 isolates from the USA and 10 isolates from France, clearly showed that M. pneumoniae is a highly uniform species and that most of the isolates could be assigned to one of the two subtypes 1 and 2. The members of one subtype differ from the other with respect to the sequence of the P1 gene, the ORF6 gene, the P65 gene, and by a typical DNA restriction fragment pattern. We observed four isolates (variants), which seemed identical by the above mentioned criteria, but did not belong to either one of the two subtypes. They showed most of the subtype 2-specific features, but differed in the sequence of the P1 gene and showed a variation in the restriction fragment pattern. The appearance of subtype 1 or 2 over the last 12 years in Germany showed a dominance of subtype 1 between 1989 and 1996 and a dominance of subtype 2 between 1997 and 1998. The variant (neither subtype 1 nor subtype 2) was only detected in 1991 and 1995 but it had no epidemiological consequences. PMID- 12635935 TI - Importance of the terminal complement components for immune defence against Candida. AB - Candida activates complement via all three pathways leading to opsonisation and anaphylaxis. The aim of the study was to investigate the influence of the terminal complement system on Candida infections. Thus, fungal cell growth, mitochondrial activity and phagocytosis by polymorphonuclear leukocytes (PMNLs) as well as specific virulence factors, such as release of secreted aspartic protease (Sap) and adherence to epithelial cells, were assessed under the influence of normal or C6/C7-depleted serum. Candida (C.) dubliniensis was used in all experiments as prototype because of its known increased expression of Saps and its strong geno- and phenotypical similarity to the most abundant Candida species C. albicans. Being exposed to sufficient quantities of complement, fungal growth decreased and phagocytosis increased but mitochondrial activities of the yeast increased as well. Concerning the virulence factors, both adhesion and especially Sap release were markedly reduced in the presence of high serum concentrations. Interestingly, at low serum concentrations some opposite effects (an augmented cell growth, a higher Sap release and a stronger adhesion) were observed. In particular, it was shown that the presence of terminal complement factors, and thus the generation of the membrane attack complex, clearly induced a higher fungal mitochondrial activation and has an effect on host defence against yeast cells by augmenting phagocytosis. PMID- 12635936 TI - Helicobacter pylori infection: CagA-specific antibodies are associated with clinical outcome, but not with HLA-class II polymorphisms of the host. AB - In order to investigate the interrelation between immunogenetic host and bacterial factors the T-cell receptor (TCR) polymorphism TCRBV6S1 A/B, HLA-DRB1 alleles and cagA status was analyzed in 380 unrelated German individuals. H. pylori infection with cagA-positive bacterial strains was significantly associated with peptic ulcer disease in the German cohort. Patients homozygous for the non-functional TCRBV6S1B allele and presenting with peptic ulcer disease showed no CagA-specific antibodies in the majority of cases. There was no association between HLA-DRB1 alleles and the CagA status of infected individuals, although certain alleles show significant association to the infection status in different populations. PMID- 12635937 TI - Molecular typing of exfoliative toxin-producing Staphylococcus aureus strains involved in epidermolytic infections. AB - Genotyping of sixteen exfoliative toxin-producing (ET-positive) strains of Staphylococcus aureus isolated in maternity units of two distant hospitals was accomplished by PFGE, ribotyping, PCR ribotyping, and prophage carriage. Three strains secreted combined ETA + ETB, and the remaining produced ETA and enterotoxin C, or TSST-1. The comparison of various genomic profiles resulted in the identification of nine genotypes. The presence of one prevailing genotype was demonstrated in each hospital. Evidence was given that the outbreak-related ET positive strains causing the skin disease pemphigus neonatorum disseminated in both the hospitals did not originate from a single source or a common ancestor. PMID- 12635938 TI - The role of intimin in the adherence of enterohaemorrhagic Escherichia coli (EHEC) O157: H7 to HEp-2 tissue culture cells and to bovine gut explant tissues. AB - Intimin, an outer membrane protein encoded by eaeA, is a key determinant for the formation of attaching and effacing (AE) lesions by enterohaemorrhagic Escherichia coli (EHEC). To investigate the role of intimin in adherence, the eaeA gene was insertionally inactivated in three EHEC O157:H7 strains of diverse origin. The absence or presence of intimin did not correlate with the extent of adhesion of mutant or wild-type O157:H7 in tissue culture and neonatal calf gut tissue explant adherence assays. Adherence of the eaeA mutants to HEp-2 cells was diffuse with no evidence of intimate attachment whereas wild-type bacteria formed microcolonies and AE lesions. Intimin-independent adherence to neonatal calf gut explants was demonstrated by eaeA mutants and wild-type strains which adhered in the greatest numbers to colon but least well to rumen tissue. These results confirm that intimin is necessary for intimate attachment and that additional adherence factors are involved in intimin-independent adherence. PMID- 12635939 TI - Rheumatoid arthritis: proposal for the use of anti-microbial therapy in early cases. AB - Rheumatoid arthritis (RA) is a chronic disease, affecting women more than men, especially in those possessing the "shared epitope" (EQK/RRAA) amino acid sequences present in HLA-DR1/4 molecules. Proteus mirabilis carries sequences showing molecular mimicry to the "shared epitope" and to type XI collagen of hyaline cartilage. Elevated levels of antibodies to P. mirabilis have been reported from 14 different countries involving 1375 RA patients and the microbe has been isolated from urine cultures of such patients. Our working hypothesis is that the disease develops as a result of repeated episodes of Proteus upper urinary tract infections. Prospective studies involving the trial of anti-Proteus measures in RA patients should be evaluated in the management of this disease. Antibiotics, high fluid intake, and fruit extracts, such as cranberry juice, have all been found to be effective in the treatment of urinary tract infections. Such measures could be used as possible additional adjuncts to the standard therapy with NSAIDs and DMARDs. PMID- 12635940 TI - Clonal analysis of B cells in the synovial membrane of patients with rheumatoid arthritis. AB - OBJECTIVE: To investigate the clonal characteristics of B cells in the synovial membranes of rheumatoid arthritis (RA) patients. METHODS: The clonality of B cells at separate sites of the synovial membrane and other tissues from RA patients were investigated by a reverse transcriptase-polymerase chain reaction with a VH framework 3 consensus primer and a subsequent single-strand conformation polymorphism analysis. RESULTS: Several dominant bands were observed in all synovial membrane samples and some of the dominants bands were common among the 2 or 3 separate regions of each synovial sample. The persistent existence of clonal B cells was observed in metachronous synovial fluid samples. CONCLUSION: Infiltrating B cells are oligoclonal and antigen-driven mechanisms may play a role in the generation of clonal B cells in RA synovium. The stable presence of B cell clones in synovial fluid suggests the involvement of these clones in the perpetuation of the chronic inflammation in RA. PMID- 12635942 TI - Shoulder joint involvement in patients with newly diagnosed rheumatoid arthritis. Prevalence and associations. AB - OBJECTIVE: To estimate the prevalence of reduced function, compared to normative values, and tenderness in the shoulder in patients with newly diagnosed rheumatoid arthritis (RA), and to investigate associated factors. METHODS: Eighty consecutive patients with RA, participating in a prospective study of early rheumatoid arthritis, were examined and assessed within one year of the onset of symptoms. RESULTS: The prevalence of tenderness from any of the shoulder joints was 50%, while 30% had decreased shoulder function, compared with age matched controls, in at least one shoulder. Both tender shoulder joints and decreased shoulder function were related to higher age and more severe disease, reflected by disability (HAQ) and the patient's assessment of pain and global disease activity. CONCLUSION: The shoulder girdle is involved early and often in rheumatoid arthritis and involvement is associated with a substantially more severe disease status. The study suggests that the shoulder should be given attention in patients with newly diagnosed rheumatoid arthritis, and that both tenderness and shoulder function should be evaluated. PMID- 12635941 TI - Regulation of nitric oxide production in osteoarthritic and rheumatoid cartilage. Role of endogenous IL-1 inhibitors. AB - OBJECTIVE: To investigate the endogenous regulation of interleukin-1 (IL-1) cytokine network in osteoarthritic (OA) and rheumatoid (RA) cartilage in relation to nitric oxide (NO) production. METHODS: Cartilage specimen obtained from OA and RA patients undergoing knee replacement surgery were studied for iNOS expression, NO and IL-1 antagonist production in tissue culture. RESULTS: OA cartilage responded to IL-1beta-stimulation with higher NO production than RA cartilage, whereas there was no difference in NO synthesis between OA and RA samples when stimulated by TNFalpha or LPS. Interleukin-1 receptor antagonist (IL-1Ra) production was higher in RA cartilage than in OA cartilage, and its production was increased by NO synthase inhibitor 1400W. CONCLUSION: IL-1beta is a potent stimulator of NO production by the iNOS pathway in RA and more pronouncedly in OA cartilage. This process is regulated by cartilage derived IL-1 antagonists, and is implicated in cartilage destruction and synovial inflammation in OA and RA joints. PMID- 12635943 TI - The assessment of fatigue in primary Sjogren's syndrome. AB - OBJECTIVE: Disabling fatigue is a prominent feature of primary Sjogren's syndrome (PSS). We evaluated a number of questionnaires for their ability to discriminate fatigue in PSS from that in other rheumatic disorders and healthy controls. METHODS: 33 female caucasian patients with PSS, 45 with rheumatoid arthritis (RA), 16 with systemic lupus erythematosus (SLE) and 30 controls completed self administered questionnaires including; Visual Analogue Scales (VAS), the Chalder Fatigue Scale (CFS), the Nottingham Health Profile (NHP) and the Medical Outcomes Short Form 36 Questionnaire (SF-36). RESULTS: All patient groups scored significantly worse than controls on the 'Energy' dimension of the NHP, the fatigue VAS and the 'Vitality' domain of the SF-36. No significant differences were observed between PSS patients and controls using the CFS. CONCLUSIONS: The NHP. VAS and SF-36 are useful in identifying fatigue in these rheumatic disorders. Further work is required to identify the characteristic features of fatigue in these conditions. PMID- 12635944 TI - Causes of death in polymyalgia rheumatica. A prospective longitudinal study of 315 cases and matched population controls. AB - OBJECTIVE: To determine causes of death in patients with pure polymyalgia rheumatica (PMR) compared to matched population controls. METHODS: In a population based study from 1987-1997, 315 patients were diagnosed with PMR. The patients were each randomly assigned four population controls, totally 1,260 controls. The date and causes of death were identified from the data files at Statistics Norway up to the end of 1997. RESULTS: A total of 65 cases (20.6%) with PMR died compared to 338 (26.8%) among the controls (mortality rate ratio (MRR) = 0.73, 95% CI 0.56-0.97, p = 0.03). No statistically significant difference was found between patients and controls with regard to mortality from coronary heart disease or stroke (MRR=0.78, 95% CI 0.52-1.18), cancer (MRR = 0.59, 95% CI 0.30-1.17), and other causes (MRR=0.75, 95% CI 0.48-1.17). CONCLUSION: The increased survival found in patients with PMR could not be explained by reduction in any particular cause of death. PMID- 12635945 TI - Disease activity and antinucleosome antibodies in systemic lupus erythematosus. AB - OBJECTIVE: To evaluate the correlation between antinucleosome antibodies and disease activity in patients with systemic lupus erythematosus (SLE). METHODS: We evaluated antinucleosome antibodies (by ELISA) in 48 SLE patients. They were divided in 2 groups: positive (Group A, nr = 18) and negative (Group B, nr=30). The groups were evaluated for antinucleosome antibodies and for clinical, humoral parameters (hemoglobin, blood cell count, urinanalysis, ESR, ANA, anti-dsDNA, anticardiolipin antibodies, LAC), and ECLAM. RESULTS: C3,C4, and hemoglobin were lower in Group A than (vs) group B (C3: 0.61 +/- 0.16 g/L vs 0.88 +/- 0.08 g/L, p < 0.001; C4: 0.086 +/- 0.03 g/L vs 0.18 +/- 0.07 g/L, p < 0.05; hemoglobin: 8.7 +/- 5.8 g/dL vs 12.7 +/- 1.44 g/dL; p < 0.02). ECLAM was higher in group A 7.56 +/- 2.19 vs group B 4.67 +/- 1.35 (p < 0.001). Urinary sediment was more altered in group A (88.8%) vs group B (33.3%; p < 0.001). CONCLUSION: We found a correlation between antinucleosome antibodies and SLE disease activity as expressed by the higher ECLAM score in group A. PMID- 12635946 TI - Hip disability and osteoarthritis outcome score. An extension of the Western Ontario and McMaster Universities Osteoarthritis Index. AB - To further develop the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC LK 3.0) for people with hip disability with or without hip osteoarthritis (OA), 52 subjects (median age 64 yrs, 35 women) answered a version of the Index with additional dimensions, twice, with a one-week interval. Reproducibility, percentage of zero scores (best possible scores), mean score of symptoms, and importance, were analyzed. This resulted in the Hip disability and osteoarthritis outcome score (HOOS LK 1.1), a 39-item questionnaire with five separate sub-scales. There were higher median scores (more symptoms) for three of HOOS sub-scales Pain, Activity limitations--sport and recreation, and Hip-related Quality of life compared to those in the WOMAC, improving the ability to assess change in patients over time. The HOOS appears to be an evaluative instrument for assessing important self-rated hip problems for people with hip disability with/without hip OA, but additional studies are needed. PMID- 12635948 TI - Scleroderma renal crisis sine scleroderma during pregnancy. AB - Renal crisis is a serious complication of systemic sclerosis. Its occurrence prior to the development of skin sclerosis is exceedingly rare. We report a patient who developed acute renal failure during pregnancy. Renal biopsy showed features compatible with scleroderma renal crisis but typical cutaneous changes were only evident 2 months after the renal episode. The relationship between pregnancy, scleroderma activity and renal crisis is discussed. PMID- 12635947 TI - Association of peripheral lymphadenopathy with anti-Ro and anti-La antibodies in a family with systemic lupus erythematosus. AB - We report here a mother and her two daughters suffering from systemic lupus erythematosus, accompanied by the presence of anti-Ro (SSA-52kD and 60kD) and anti-La (SSB) antibodies, and peripheral lymphadenopathy as the first manifestation of the disease. The expression of DR3 and B8 in all three affected family members suggests familiar predisposition to SLE. The differential diagnosis and clinical course are discussed. PMID- 12635949 TI - Bone scintigraphy using Tc-99m DPD and F18-FDG in a patient with SAPHO syndrome. AB - Synovitis (inflammatory arthritis), acne (pustulosa), palmoplantar pustulosis, hyperostosis and osteitis (bland osteomyelitis) are symptoms forming the acronym SAPHO. We present the case of a 48-year old man with sterno-costo-clavicular hyperostosis and typical pustulosis palmaris. We performed Tc-99m DPD serial bone scanning to monitor the course of disease and to assess therapeutic efficiency. Control bone scans in 2001 showed minor compromises of the ribs and diminished disease activity on collarbones and the sternum after medication with non steroidal anti-inflammatory drugs (NSAIDs) and bisphosphonates. F-18 FDG PET presented synovial inflammation in the left sterno-clavicular joint but no relevant tracer uptake on clavicles or breastbones. In case of diagnostic doubts F-18 FDG PET could be recommended in order to discriminate bland osteomyelitis from bacterial osteomyelitis or from bone malignancy when SAPHO-syndrome is assumed. PMID- 12635950 TI - Tanker safety and coastal environment: Prestige, Erika, and what else? PMID- 12635951 TI - Kinetics of benzene adsorption onto activated carbon. AB - An activated carbon bed adsorption process is influenced by the adsorbents' characteristics, volatile organic compound (VOC) characteristics, and process conditions. In the literatures, the adsorption processes of the adsorbents and VOCs were usually considered to be in equilibrium. In this study, the VOC adsorption processes by activated carbon were considered to be a kinetic process, i.e. they are not in equilibrium. Then, isothermal adsorption curves and a small column experiment were simulated. PMID- 12635952 TI - Ligand exchange rate of metal-NOM complexes by EDTA. AB - Metal complexation by natural ligands is important for metal transport and distribution in surface and ground water. The goal of the work was to study the ligand exchange rate for two important metal ions in natural aquatic systems (Al, Fe) was determined using EDTA and natural organic matter (NOM) of humic type as ligands. After adding EDTA to a solution containing metal-NOM complexes, these complexes dissociated and metal-EDTA complexes were formed. Metal-NOM complexes were separated from metal-EDTA complexes with the help of size-exclusion chromatography and detected by on-line inductively coupled plasma-mass spectrometry (ICP-MS). Injecting the samples into the system over time after addition of EDTA allowed us to measure the rate of the exchange of NOM by EDTA. The experiments could be well described with a first-order rate law assuming that the dissociation of the metal-NOM complexes is the rate-determining step. The exchange rate of Fe was found to be faster than that of Al. This corresponds well with the exchange rate of water molecules from the coordination sphere of the metal ions, which is also faster for Fe than for Al. Furthermore, the UV and the fluorescence signal of the chromatograms were measured. The results indicate that no disaggregation of NOM molecules took place, although about 75-85% of the aggregate-forming metal ions exchanged NOM by EDTA in their coordination sphere. This suggests clearly the fundamental role of NOM in colloidal transport of metals and in their bioavailability. PMID- 12635953 TI - Acetochlor as a soil pollutant. AB - Acetochlor is a widely used herbicide all over the world. Similarly to other organic pollutants, the environmental fate of the acetochlor is strongly related to its adsorption properties. Static adsorption equilibrium measurements were carried out at 25 degrees C on different types of Hungarian soils characterized by varying amounts of organic matter and pH values. Isotherms obtained under different conditions, as well as on various soils, exhibit a similar shape, thus indicating a two-step adsorption process. The plots cannot be interpreted according to the classes of isotherms suggested by Giles. The adsorption coefficients (K) were estimated from the initial slope of the curves. These values were determined not only by the type of the soil, but also by the composition of the aqueous media. The organic matter adsorption coefficients (Kom) were also calculated and they were approximately identical for soils of high organic matter. Due to the low value of the adsorption coefficients, the acetochlor is a rather mobile pollutant of the soil posing a potential danger to the aquatic environment. For soils with high organic content, the Kom values are similar to each other and, due to the significant coverage of the inorganic surfaces, the adsorption is controlled by the solute-organic substance interactions. The adsorption of weakly polar compounds, however, is remarkable even on those soils where the organic content is low. In this case, the binding is governed by the solute-inorganic surface interactions. This conclusion is nicely proved by the adsorption of the acetochlor on quartz. According to our hypothesis, the second part of the adsorption step is controlled by the solute adsorbed solute interactions. The role of the organic matter in this region of the isotherm is probably negligible. As the organic matter content of the soils plays an important role in the acetochlor adsorption, humic substances must have a strong influence upon the transport of this compound. Experiments to obtain adsorption isotherms of further pesticides and the development of a quantitative model are in progress. PMID- 12635954 TI - Long-term variation in surface ozone and its precursors in Athens, Greece: a forecasting tool. AB - INTENTION, GOAL, SCOPE, BACKGROUND: Photochemical pollution is a very complex process involving meteorological, topographic, emission and chemical parameters. The most important chemical mechanisms involved in the atmospheric process have already been identified and studied. However, many unknown parameters still exist because of the large number of participating chemical reactions. OBJECTIVE: The present study investigates the processes involved in the photochemical pollution effect of an urban station located in the greater area of the Athens basin and gives a plausible explanation for the different seasonal ozone development between that station and another rural one. Furthermore, the distribution of the mean monthly surface ozone observed at the urban station during 1987-2001 is examined in order to create a relevant forecasting tool. METHODS: Averaged hourly data of O3 and NOx observations monitored at the above mentioned stations, during 1987-2001, have been used in order to derive the daytime (7:00-15:00) values. Trajectories calculated by using a 2D-trajectory code and meteorological data, during the period 1988-1996, have also been used. RESULTS AND DISCUSSION: At the urban station, the percentage negative trend of NO and NOx data in winter and summer is higher than that in spring and autumn, while the percentage ozone trend is maximum in the summer. On the contrary, the negative surface ozone trend at the rural station exhibits a minimum in summer and a maximum in autumn and winter. The mean seasonal wind-rose for the selected months shows that the northward wind flow dominates during June, the month of the lowest negative ozone trend in the rural station. Finally, the development of the forecasting tool shows that the mean monthly surface ozone data during the period (1987-2001) demonstrates a semi-log distribution. CONCLUSIONS: Air transport effect on the air pollution of the rural station (not blocked by mountains) is deduced as a possible reason for the different seasonal ozone development observed between the rural and the urban station. Finally, the discrepancies between the theoretical probabilities deduced by the model and the empirical ones appear to be very small, and the corresponding correlation coefficient is 0.99. RECOMMENDATION AND OUTLOOK: However, to interpret the aforementioned statistical results about the negative trends in ozone and its precursors, additional parameters can be taken into account. Changes in NOx concentrations, for instance, can result not only from changes in emissions or meteorological conditions. There might also be a contribution through changes in the atmospheric composition. A study of the contribution of changes in atmospheric composition to trends of observed NOx concentrations requires that a series of steps be taken (removal of meteorological influence in the time series, calculation of trends in OH concentrations, etc.). PMID- 12635955 TI - The influence of meteorological conditions and stringent emission control on high TSP episodes in Istanbul. AB - Istanbul has faced serious air pollution problems since the mid-80s. This is mainly due to particulate air pollution coming from poor quality lignite in areas, which are heavily populated and industrialized. As a consequence of severe air pollution problems, stringent control on the emissions in the city started in the year of 1994. In this work, in order to study the relationship between emissions and meteorological conditions, an assessment of air pollution episodes and air pollution potential in the city is presented for the terms at the changed emission schedule as the influence of an emission reduction strategy. The influence of meteorological conditions on the TSP (total suspended particulates) levels is considered for two consecutive winter periods. On this occasion, the city has faced different TSP levels and episode characteristics depending on stringent emission reductions covering the banned, poor-quality lignite and fuel switching. For this purpose, climatological conditions and air quality analyses were performed. PMID- 12635957 TI - Dehalogenation potential of municipal waste incineration fly ash. I. General principles. AB - BACKGROUND, AIMS AND SCOPE: It is well known that the fly ash from filters of municipal waste incinerators (MWI-FA) shows dehalogenation properties after heating it to 240-450 degrees C. However, this property is not general, and fly ash samples do not possess dehalogenation ability at all in many cases. Fly ash has a very variable composition, and the state of the fly ash matter therefore plays the decisive role. In the present paper, the function of important components responsible for the dehalogenation activity of MWI-FA is analysed and compared with the model fly ash. METHODS: With the aim of accounting for the dehalogenation activity of MWI-FA, the following studies of hexachlorobenzene (HCB) dechlorination were performed: The role of copper in dehalogenation experiments was evaluated for five types of metallic copper. The gasification of carbon in MWI-FA was studied in the 250-350 degrees C temperature range. Five different kinds of carbon were used, combined with conventional Cu(o) and activated nanosize copper powder. The dechlorination experiments were also carried out with Cu(II) compounds such as CuO, Cu(OH)2, CuCl2 and CuSO4. The results were discussed from the standpoint of thermodynamics of potential reactions. Based on these results, the model of fly ash was proposed, containing silica gel, metallic copper and carbon. The dechlorination ability of MWI-FA and the model fly ash are compared under oxygen-deficient atmosphere. CONCLUSIONS: The results show that, under given experimental conditions, copper acts in the dechlorination as a stoichiometric agent rather than as a catalyst. The increased surface activity of copper enhances its dechlorination activity. It was found further that the presence of copper leads to a decrease in the temperature of carbon gasification. The cyclic valence change from Cu(o) to Cu+ or Cu2+ is a prerequisite for the dehalogenation to take place. RECOMMENDATION AND OUTLOOK: Thermodynamic analysis of the dechlorination effect, as well as the comparison of dechlorination pathways on MWI-FA and model fly ash, can provide a deeper understanding of the studied reaction. PMID- 12635956 TI - Ferrioxalate-mediated photodegradation and mineralization of 4-chlorophenol. AB - INTENTION, GOAL, SCOPE, BACKGROUND: Advanced oxidation processes are powerful methods which are capable of transforming refractory, nonbiodegradable and/or toxic organic compounds into harmless end products such as carbon dioxide and water. However, one commen problem of all advanced oxidation processes is the high demand of electrical energy for ultraviolet lamps, which causes high operational costs. Minimization of the required irradiation time, and therefore the energy consumption, by optimization of other reaction conditions such as catalyst-oxidant type and concentration, pH, temperature, pollutant/oxidant ratio etc., therefore continues to gain importance. OBJECTIVE: The main objective of this study was the minimization of the required irradiation time through optimization of the use of a newly patented catalyst, ferrioxalate, and also to compare the performance of this catalyst with the performance of other AOPs. METHODS: Oxidation of 4-chlorophenol by photo-Fenton process using potassium ferrioxalate as a mediator was studied in a lab scale photoreactor. The influence of parameters such as hydrogen peroxide and ferrioxalate concentrations, initial pH, power-output, oxalate/iron ratio and different iron sources was evaluated. An upflow photoreactor equipped with a 1000 Watt high-pressure mercury vapour lamp and operating in a recirculation mode was used during photodegradation experiments. The extent of the reduction of 4-chlorophenol, Total Organic Carbon and Chemical Oxygen Demand was used to evaluate the photodegradation reaction. RESULTS AND DISCUSSION: The optimum pH range observed was found to be 2.7-3. The efficiency of 4-chlorophenol oxidation increased with increasing concentrations of hydrogen peroxide and ferrioxalate, reaching a plateau after the addition of 10 and 0.072 mM of those reagents, respectively. Using an Oxalate/iron ratio of 12 was 18% less efficient than using a ratio of 3:1. The efficiency increased with increasing radiation power. However, this increase was not linear. The UV/ferrioxalate/H2O2 process, by which complete mineralization of 100 mg l(-1) 4 chlorophenol was achieved in 20 min of total reaction time, was the most efficient process among the alternatives applied. CONCLUSIONS: The use of ferrioxalate as the catalyst was found to be more efficient than the use of Fe(II) and Fe(III) iron species. It was possible to completely mineralize 4 chlorophenol. RECOMMENDATION AND OUTLOOK: The results of this study demonstrate that the ferrioxalate-mediated degradation of 4-chlorophenol requires less irradiation times than other advanced oxidation processes. There are mainly 19 phenol isomers and other toxic and nonbiodegradable organic compounds. We recommend that similar studies should be performed on many such compounds in order to attain a clear understanding of the performance of this catalyst. Because of its light sensitivity, this catalyst should be used immediately after its preparation. The use of low pressure mercury vapour lamps in this process should also be considered, since low power outputs may be enough for the process. PMID- 12635958 TI - Minimum specific cost control of technological processes realized in a living objects-containing microenvironment. AB - The purpose of the present work is to work out an approach for the development of software and the choice of hardware structures when designing subsystems for automatic control of technological processes realized in living objects containing limited space (microenvironment). The subsystems for automatic control of the microenvironment (SACME) under development use the Devices for Air Prophylactic Treatment, Aeroionization, and Purification (DAPTAP) as execution units for increasing the level of safety and quality of agricultural raw material and foodstuffs, for reducing the losses of agricultural produce during storage and cultivation, as well as for intensifying the processes of activation of agricultural produce and industrial microorganisms. A set of interconnected SACMEs works within the framework of a general microenvironmental system (MES). In this research, the population of baker's yeast is chosen as a basic object of control under the industrial fed-batch cultivation in a bubbling bioreactor. This project is an example of a minimum cost automation approach. The microenvironment optimal control problem for baker's yeast cultivation is reduced from a profit maximum to the maximization of overall yield by the reason that the material flow oriented specific cost correlates closely with the reciprocal value of the overall yield. Implementation of the project partially solves a local sustainability problem and supports a balance of microeconomical, microecological and microsocial systems within a technological subsystem realized in a microenvironment maintaining an optimal value of economical criterion (e.g. minimum material, flow-oriented specific cost) and ensuring: (a) economical growth (profit increase, raw material saving); (b) high security, safety and quality of agricultural raw material during storage process and of food produce during a technological process; elimination of the contact of gaseous harmful substances with a subproduct during various technological stages; (c) improvement of labour conditions for industrial personnel from an ecological point of view (positive effect of air aeroionization and purification on human organism promoting strengthened health and an increase in life duration, pulverent and gaseous chemical and biological impurity removal). An alternative aspect of a controlled living microenvironment forming is considered. PMID- 12635959 TI - Survey of dioxin sources in the Baltic Region (extended summary). AB - The present paper summarises the results of the project: 'Survey of Anthropogenic Sources of Dioxins and Furans in the Baltic Region'. As a part of the project, inventories have been carried out in Estonia, Latvia, Lithuania and Poland by applying the toolkit for quantification of dioxin and furan releases developed by UNEP Chemicals. The main route of direct releases to the environment is emission to air. Total emission to air from Poland was estimated at 490 (88-1,300) g I TEQ/year, whereas the emissions from Estonia, Latvia and Lithuania were estimated as being 14 (2.4-54), 23 (2.6-63) and 17 (2.6-38) g I-TEQ, respectively. In general, the uncertainty on the estimates is very high, and recommendations regarding further development of the inventories have been made, and measures for reducing the releases have been provided. PMID- 12635960 TI - Part I: Integrated water quality management: river basin approach. Geochemical techniques on contaminated sediments--river basin view. AB - The big flood in the upper Elbe River catchment area has revealed a wide spectrum of problems with contaminated sediments. So far, an effective strategy for managing contaminated sediments on a river basin scale is still missing and it seems that not much has been learned from the lessons received during the last decade. In the following overview, special emphasis is given to the utilization of geochemically-based techniques for sediment remediation, which can be applied in different parts of a river basin. The examples presented here are mostly from the Elbe River catchment area. In general, new technical problem solutions need a set of practical process knowledge that uses a wide range of simulation techniques, as well as models in different spatial and temporal scales. The evaluation of recent flood events clearly demonstrates the importance of chemical expertise in the decision-making process for the sustainable development in river basins. PMID- 12635961 TI - General prevention and risk minimization in LCA: a combined approach. AB - Methods for life cycle assessment of products (LCA) are most often based on the general prevention principle, as opposed to the risk minimization principle. Here, the desirability and feasibility of a combined approach are discussed, along with the conditions for elaboration in the framework of LCA methodology, and the consequences for LCA practice. A combined approach provides a separate assessment of above and below threshold pollution, offering the possibility to combat above threshold impacts with priority. Spatial differentiation in fate, exposure, and effect modelling is identified to play a central role in the implementation. The collection of region-specific data turns out to be the most elaborate requirement for the implementation in both methodology and practice. A methodological framework for the construction of characterization factors is provided. Along with spatial differentiation of existing parameters, two newly introduced spatial parameters play a key role: the sensitivity factor and the threshold factor. The practicability of the proposed procedure is illustrated by an example of its application. Providing a reasonable data availability, the development of separate LCA characterization factors for the respective assessment of pollution levels above and below environmental threshold values seems to be a feasible task that may add to LCA credibility. PMID- 12635962 TI - Practitioner review: outcomes for parents and children following non-traditional conception: what do clinicians need to know? AB - BACKGROUND: The birth of the first 'test-tube' baby in 1978 has led to the creation of family types that would not otherwise have existed. METHODS: This paper examines research on the psychological outcomes for parents and children in assisted reproduction families with particular attention to the issues and concerns that have been raised by creating families in this way. RESULTS: Parents of children conceived by assisted reproduction appear to have good relationships with their children, even in families where one parent lacks a genetic link with the child. With respect to the children themselves, there is no evidence of cognitive impairment in singleton children born at full term as a result of assisted reproduction procedures, although the findings regarding the cognitive development of intracytoplasmic sperm injection (ICSI) children are currently unclear. In relation to socioemotional development, assisted reproduction children appear to be functioning well. CONCLUSIONS: Few studies have included children at adolescence or beyond, and little is known about the consequences of conception by assisted reproduction from the perspective of the individuals concerned. In addition, there are some types of assisted reproduction family, such as families created through a surrogacy arrangement or through embryo donation, about whom little is known at all. Although existing knowledge about the impact of assisted reproduction for parenting and child development does not give undue cause for concern, there remain a number of unanswered questions in relation to children born in this way. PMID- 12635963 TI - Maternal adjustment, parenting and child behaviour in families of school-aged twins conceived after IVF and ovulation induction. AB - BACKGROUND: Previous studies that have examined the long-term effects of infertility and assisted reproductive technology on parenting and child behaviour in families with twins have suffered from methodological problems. This study compared measures of parental adjustment, parenting and child behaviour in families with 5-year-old twins who were conceived after in vitro fertilisation (IVF) or ovulation induction (OI) with families whose twins were naturally conceived (NC). METHODS: The families who conceived via IVF/OI (N = 121) were identified from an epidemiological study of twins and matched to families who were conceived naturally (N = 121) on the basis of eleven child and family variables. Mothers were interviewed in their homes for the study. RESULTS: No significant differences were observed between the IVF/OI families and the NC families on measures of parental adjustment or parent and teacher ratings of the twins' behaviour. IVF/OI mothers and their partners agreed with each other about discipline more than NC couples, but otherwise no other differences in parenting were found. CONCLUSIONS: Overall, this study provides evidence that families who conceive twins following IVF/OI are functioning well and that the experience of fertility treatment does not lead to long-term difficulties for parents or children. PMID- 12635964 TI - Twins as a natural experiment to study the causes of mild language delay: I: Design; twin-singleton differences in language, and obstetric risks. AB - BACKGROUND: Twins tend to lag behind singletons in their language development, but the causes were unknown. The possibilities suggested include obstetric complications, twin-specific features, and postnatal differences in family interaction. The present study was designed to pit these alternatives against one another as possible causal influences. METHOD: The Avon Longitudinal Study of Parents and Children (ALSPAC) was used to identify the 116 twin pairs (of whom 96 participated) and 114 pairs of singletons (of whom 98 participated) whose ages were no more than 30 months apart. The McArthur Communicative Development Inventory was completed at 20 months, and the Pre-School Language Scales (PLS-3), and the McCarthy Scales of Children's Abilities at 36 months. Obstetric and perinatal complications were assessed on the basis of detailed systematic parental reports, together with a systematic coded abstraction of all medical records dealing with pregnancy and the neonatal period. Family background details were assessed from parental reports, and the primary carer's verbal functioning was assessed by the Mill Hill Vocabulary Scale. Congenital anomalies were assessed using the method of Waldrop and Halverson. RESULTS: The language of twins was 1.7 months below that of singletons at 20 months and 3.1 months at 3 years. The verbal cognitive score of twins was about half a standard deviation lower than that of singletons. The twin-singleton differences in language level were found tobe unassociated with obstetric/perinatal features as assessed from both parental reports and medical records, to birthweight or gestation, to birthweight discrepancy within the twin pair, or to congenital anomalies. CONCLUSION: It is concluded that obstetric/perinatal features do not account for the slower language development in twins as compared with singletons, within a sample born after at least 33 weeks gestation. PMID- 12635965 TI - Twins as a natural experiment to study the causes of mild language delay: II: Family interaction risk factors. AB - BACKGROUND: Twins tend to lag behind singletons in their language development, but the causes were unknown. METHOD: Ninety-six twin pairs from the Avon Longitudinal Study of Parents and Children (ALSPAC), for whom birth was after at least 33 weeks of gestation, were compared with 98 pairs of singletons, no more than 30 months apart in age. Parental qualities and family interaction were assessed through standardised questionnaires and interviews and both structured and unstructured observations in the home at 20 months and 36 months. The possible causal role of postnatal family influences was assessed through five criteria: i) the feature had to differ between twins and singletons; ii) individual differences in that feature had to relate to individual differences in language level within the sample of singletons and of twins; iii) the feature as measured at 20 months had to predict language as assessed at 36 months; iv) that had to apply after controlling for language level at 20 months; and v) introduction of the predictive feature into an overall model had to obliterate the twin-singleton difference in language level. RESULTS: Patterns of parent child interaction and communication met these five criteria. The maternal factors all concerned aspects of interaction that were broadly concerned with communication: encouraging the child to speak, providing elaborating comments, engaging in reading to the child and talking about the story and its illustrations. The HOME inventory findings provided similar findings with respect to responsiveness, involvement and level of experiences involved. Family features that might have been influential, but which were not, included parental depression, breast-feeding, family size, and style of sibling interaction. CONCLUSION: Patterns of parent-child interaction and communication within the normal range have environmentally mediated effects on language and account for twin-singleton differences in language developmently. The results indicate the value of a natural experiment in testing competing causal hypotheses, and show the role of environmental factors as influences on language variations within the normal range, for both twins and singletons. PMID- 12635966 TI - Association of ADHD and conduct disorder--brain electrical evidence for the existence of a distinct subtype. AB - BACKGROUND: To evaluate the impact of psychopathological comorbidity with oppositional defiant/conduct disorder (ODD/CD) on brain electrical correlates in children with attention deficit hyperactivity disorder (ADHD) and to study the pathophysiological background of comorbidity of ADHD+ODD/CD. METHOD: Event related potentials (ERPs) were recorded during a cued continuous performance test (CPT-A-X) in children (aged 8 to 14 years) with ICD-10 diagnoses of either hyperkinetic disorder (HD; n = 15), hyperkinetic conduct disorder (HCD; n = 16), or ODD/CD (n = 15) and normal children (n = 18). HD/HCD diagnoses in all children were fully concordant with the DSM-IV diagnosis of ADHD-combined type. ERP microstates, i.e., time segments with stable brain electrical map topography were identified by adaptive segmentation. Their characteristic parameters and behavioral measures were further analyzed. RESULTS: Children with HD but not comorbid children showed slower and more variable reaction times compared to control children. Children with HD and ODD/CD-only but not comorbid children displayed reduced P3a amplitudes to cues and certain distractors (distractor-X) linked to attentional orienting. Correspondingly, global field power of the cue CNV microstate related to anticipation and preparation was reduced in HD but not in HCD. Topographical alterations of the HD occurred already in the cue-P2/N2 microstate. In sum, the comorbid group was less deviant than both the HD-group and the ODD/CD-group. CONCLUSIONS: The findings suggest that HD children (ADHD combined type without ODD/CD) suffer from a more general deficit (e.g., suboptimal energetical state regulation) including deficits of attentional orienting and response preparation than just a responseinhibitory deficit, backing the hypothesis of an involvement of a dysregulation of the central noradrenergic networks. The results contradict the hypothesis that ADHD+ODD/CD represents an additive co-occurrence of ADHD and ODD/CD and strongly suggest that it represents a separate pathological entity as considered in the ICD-10 classification system, which differs from both HD and ODD/CD-only. PMID- 12635967 TI - Classes of disruptive behaviour in a sample of young elementary school children. AB - BACKGROUND: The objectives were to (1) classify young children to groups differing in disruptive behaviour, (2) determine whether the patterns of disruptive behaviour in these groups are in accordance with the conceptualisation of disruptive disorders as described in DSM-IV, and (3) optimise the classification of children in groups. METHODS: Disruptive behaviour of 636 seven year-old elementary schoolchildren was assessed with the CBCL/4-18. Using CBCL items rated as very consistent with DSM-IV categories Conduct Disorder, Oppositional Defiant Disorder and Attention-Deficit/Hyperactivity Disorder, latent class analyses were applied to identify classes of children differing in patterns of disruptive behaviour. RESULTS: Three classes were identified: (1) high levels of oppositional defiant problems (ODD problems) and attention deficit/hyperactivity problems (ADH problems) and intermediate levels of Conduct problems; (2) intermediate ODD problems and ADH problems and low levels of Conduct problems; (3) low levels on all disruptive behaviours. CONCLUSIONS: No classes were identified in which children had symptoms of only Conduct problems, only ODD problems or only ADH problems. Covariates (socio-economic status, gender, parenting stress) improved the classification of children. The findings are discussed in terms of implications for classification, identification of children at risk, prevention and treatment of disruptive behaviour in young children. PMID- 12635968 TI - A growth curve analysis of parent training outcomes: examining the influence of child risk factors (inattention, impulsivity, and hyperactivity problems), parental and family risk factors. AB - BACKGROUND: Parent training is one of the most effective treatments for young children with conduct problems. However, not every family benefits from this approach and approximately one-third of children remain in the clinical range at follow-up assessments. Little is known about factors affecting treatment outcome for young children. METHOD: Hierarchical linear modeling methods were used to examine the effects of child attentional risk factors (inattention, impulsivity and hyperactivity problems), parental and familial risk factors upon the efficacy of a parent training program to decrease boys' conduct problems. Mothers of 81 boys, four to seven years of age, exhibiting conduct problems attended a parent training program (The Incredible Years) which lasted 22 to 24 weeks. Treatment effectiveness was assessed at one month and one year post treatment by means of independent home observations, parent and teacher reports. RESULTS: Results indicated significant decreases in observations of mothers' negative parenting interactions with their children and decreases in their conduct problems according to mother reports and independent observations at home. Boys with elevated ratings of attentional problems in addition to conduct problems showed similar benefits from the parent training program as the boys who did not have attentional problems. CONCLUSION: This study suggests that parent training is equally effective for boys with both conduct problems and attentional problems as it is for boys with conduct problems without these attentional problems. In fact, boys rated in the Borderline or Clinical range for attentional problems evidenced a greater decrease in externalizing behavior problems as rated by their mothers. Surprisingly, the predictor variables of depression, stress, and socioeconomic status were not significant contributors at either initial status or growth over time whether outcomes were measured by child conduct problems or parenting interactions. PMID- 12635969 TI - Fifteen, female and stressed: changing patterns of psychological distress over time. AB - BACKGROUND: Despite a widespread view that the mental health of young people has deteriorated, the evidence base is limited by the lack of comparable datasets over time, and their capacity to test specific hypotheses about the causes of such change, in this case those particularly affecting young females. METHOD: Two cohorts of 15-year-olds in the West of Scotland, surveyed in 1987 and 1999, were compared, using the 12-item version of the General Health Questionnaire to measure psychological distress (GHQ caseness, cut-off 2/3), together with items and indices of personal and performance worries. RESULTS: Between 1987 and 1999, GHQ caseness increased significantly for females (from 19% to 33%), but not males (13% to 15%), a change particularly experienced by females from non-manual and skilled manual backgrounds. With one notable exception (unemployment), most worries also increased for both sexes, a gender gap emerging in respect of worries about school performance, females worrying more. While the effect of personal worries (e.g., looks and weight) on GHQ caseness persisted over time in both sexes, that of performance worries only emerged for females in 1999. Using survey date as an indicator, a relationship between proximity to exams and GHQ caseness was also only found among females in 1999. CONCLUSION: The increase in levels of psychological distress among young females over this period may be explained by an increase in educational expectations, which together with more traditional concerns about personal identity, appear to have elevated levels of stress, with adverse consequences for mental health. PMID- 12635970 TI - Predictors of self-reported problem behaviours in Turkish immigrant and Dutch adolescents in the Netherlands. AB - BACKGROUND: Although many studies have compared psychopathology in different cultures, not much is known about factors that contribute to the observed differences. METHODS: We compared self-reported emotional and behavioural problems in 363 Turkish immigrant and 1098 Dutch adolescents in the Netherlands and we evaluated the contribution of adolescent, parent, family and stress related factors to the observed ethnic differences. Data were drawn from the Dutch version of the Youth Self-Report (YSR), as well as from Dutch and Turkish parental questionnaires. RESULTS: Turkish girls scored higher on four of the eight YSR syndrome-scales, on the Internalising broadband scale and on total problems than Dutch girls. Turkish boys scored higher on three syndrome scales and on the Internalising scale, but scored less on Delinquent Behaviour than their Dutch peers. Ethnic differences for both sexes were most pronounced on the Withdrawn and Anxious/Depressed scales. Socio-economic measures, in particular education of the parents, contributed most to the explanation of ethnic differences on the Somatic Complaints scale for girls and Social Problem and Internalising scales for boys. On most scales, however, ethnic differences could not be explained by other factors. The distribution of some factors appeared to be more favourable (i.e., less frequent) for Turkish than for Dutch youths, such as referral of family members to mental health services. CONCLUSIONS: Low educational levels of the parents play an important, yet not an exclusive role in explaining cross-cultural differences in emotional and behavioural problems in adolescents. In particular, differences in Withdrawn and Anxious/Depressed behaviour could not be explained by non-cultural factors. This study offers starting-points for future research on cultural-specific predictors of psychopathology in immigrants. PMID- 12635971 TI - Empathy and response to distress in children with Down syndrome. AB - BACKGROUND: Most studies of empathy have focused on young children, and those who are typically developing. Thus, we know little about the emergence and manifestation of empathy in non-normally developing children. METHOD: Empathy and response to distress in others were examined in 30 children with Down syndrome, 22 children with nonspecific etiologies of mental retardation, and 22 typically developing children. RESULTS: Results indicated that compared to the other children, children with Down syndrome responded to distress in others by looking to them more, and offering more comfort in the form of prosocial responses. However, in a hypothetical empathy situation, children with Down syndrome were less likely to feel the same emotion as the protagonist than were the typical children. Children with Down syndrome differed from the children with nonspecific mental retardation only in their response to distress in others. The children with nonspecific mental retardation were more similar to than different from the MA-matched typical children. CONCLUSIONS: These results suggest some etiology associated differences in empathy and response to distress in children with mental retardation. PMID- 12635972 TI - The influence of children's self-report trait anxiety and depression on visual search for emotional faces. AB - BACKGROUND: This study presents two experiments that investigated the relationship between 7- and 10-year-olds' levels of self-report trait anxiety and depression and their visual search for threatening (angry faces) and non threatening (happy and neutral faces) stimuli. METHOD: In both experiments a visual search paradigm was used to measure participants' reaction times to detect the presence or absence of angry, happy or neutral schematic faces (Experiment 1) or cartoon drawings (Experiment 2). On target present trials, a target face was displayed alongside three, five or seven distractor items. On target absent trials all items were distractors. RESULTS: Both experiments demonstrated that on target absent (but not present) trials, increased levels of anxiety produced significantly faster search times in the angry face condition, but not in the neutral condition. In Experiment 2 there was some trend towards significance between anxiety and searches for happy faces in absent trials. There were no effects of depression on search times in any condition. CONCLUSION: The results support previous work highlighting a specific link between anxiety and attention to threat in childhood. PMID- 12635973 TI - Children's colour choices for completing drawings of affectively characterised topics. AB - BACKGROUND: This study was designed to explore whether or not children systematically use particular colours when completing drawings of affectively characterised topics. METHOD: Three hundred and thirty 4-11-year-old children were subdivided into three conditions, colouring in a drawing of a man, a dog, or a tree, respectively. The children completed two test sessions in counterbalanced order. In one session, children rated and ranked ten colours in order of preference. In the other session, children completed three colouring tasks in which they had to colour in three identical figures but which had been given different affective characterisations: a neutrally characterised figure, a figure characterised as nasty, and a figure characterised as nice. RESULTS: It was found that, in all age groups and for all topics, the children used their more preferred colours for the nice figures, their least preferred colours for the nasty figures, and colours rated intermediately for the neutral figures. It was also found that, in all age groups and for all topics, black tended to be the most frequently chosen colour for colouring in the drawings of the negatively characterised figures. By contrast, primary colours were predominantly selected for the neutral figure, while a wide range of mainly primary and secondary colours were chosen for colouring in the nice figure. CONCLUSIONS: These results suggest that children are able to alter systematically their use of colour during picture completion tasks in response to differential affective topic characterisations, and that even very young children are able to use colours symbolically. PMID- 12635974 TI - Joint attention training for children with autism using behavior modification procedures. AB - BACKGROUND: Deficits in joint attention are considered by many researchers to be an early predictor of childhood autism (e.g., Osterling & Dawson, 1994) and are considered to be pivotal to deficits in language, play, and social development in this population (Mundy, 1995). Although many researchers have noted the importance of joint attention deficits in the development of children with autism (e.g., Mundy, Sigman, & Kasari, 1994) and have called for intervention strategies (e.g., Mundy & Crowson, 1997), few studies have attempted to target joint attention. In this study, joint attention behaviors were taught to children with autism using a behavior modification procedure. METHODS: A multiple-baseline design was implemented to evaluate intervention effects. The following target behaviors were included in the intervention: 1) Responding to showing, pointing, and gaze shifting of adult; 2) Coordinated gaze shifting (i.e., coordinated joint attention); and 3) Pointing (with the purpose of sharing, not requesting). Generalization to setting and parent, follow-up sessions, and social validation measures were also analyzed. RESULTS: Joint attention behaviors were effectively trained and targeted behaviors generalized to other settings. In addition, positive changes were noted by naive observers using social validation measures. CONCLUSIONS: Integrating joint attention training into existing interventions may be important for children with autism. In addition, training parents in these techniques may help to maintain joint attention skills outside of the treatment setting. PMID- 12635975 TI - Childhood voice disorders: ambulatory evaluation and operative diagnosis. PMID- 12635976 TI - Frequency and timing of symptoms in infants screened for sepsis: effectiveness of a sepsis-screening pathway. AB - To determine the frequency and timing of symptoms and to evaluate the effectiveness of a sepsis-screening pathway in term and near-term infants, data were collected prospectively for a period of 1 year from December 1, 2000, to November 30, 2001. Results confirmed that a sepsis-screening pathway using a combination of at least 2 serial complete blood cell count and C-reactive protein measurements in both symptomatic and asymptomatic infants is a safe, simple strategy that prevents unnecessary treatment of infants with risk factors with antibiotics. However, most infants with presumed or suspected early-onset sepsis are symptomatic. Routine treatment of asymptomatic infants with risk factors or prior treatment with intrapartum antibiotics is unnecessary. A combined approach of screening in the presence of risk factors and /or symptoms of sepsis and adequate follow-up for infants discharged at less than 72 hours of age may help reduce unnecessary treatment of infants with antibiotics. PMID- 12635977 TI - Apparent life-threatening episodes as the first manifestation of epilepsy. AB - Apparent life-threatening episodes (ALTE) can be the first manifestation of an epileptic seizure in children. However, the diagnosis of ALTE is generally delayed because the interictal EEG often appears normal and the presence of gastroesophageal reflux may mislead the diagnosis of the epileptic manifestation. We report 6 children with epilepsy who presented an ALTE as their first seizure manifestation. The retrospective analysis of our cases demonstrated that ictal EEGs, when available, appeared to be the most helpful test to confirm the diagnosis, although an abnormal-appearing cerebral imaging was also contributive in the patients when only interictal EEGs were available. PMID- 12635979 TI - Solid tumors in the neonatal period. AB - From 1972 to 2000, 123 patients with solid tumors whose complaints had started in the first 28 days of life were retrospectively evaluated. Fifty-five patients were diagnosed in the first 28 days and 68 patients were diagnosed after 28 days. In the former group, 85.5% of patients had symptoms in the first day of life. In the latter group, 77.9% had the onset of symptoms in the first day. Tumor subgroups in the neonatal period included teratoma (34), neuroblastoma (11), rhabdomyosarcoma (3), Wilms tumor (1), and retinoblastoma (3), and the others (3). Three patients had other, less common tumors. In the second group the numbers were the following: for teratoma (32), neuroblastoma (15), germ cell tumors other than teratomas (8), rhabdomyosarcomas (4), the other soft tissue sarcomas (3), Wilms tumor (1), retinoblastoma (1), and other, rare tumors (4). There were 22 malignant tumors in the first group, and 44 in the second group. Fourteen patients in the first group died in the early postoperative period or with progressive disease. Nineteen of 44 patients died in the second group. Overall survival rates were 24.9% and 51.6% in first and second groups, respectively (p = 0.015). Event-free survival rates were 14.7% and 47.7% in these groups, respectively (p = 0.0063). This is the first report comparing clinical features and prognosis of tumors diagnosed in the first 28 days of the life with those diagnosed after 28 days. The prognosis was worse in infants diagnosed in the first 28 days of life. PMID- 12635978 TI - Role of imaging in the diagnosis of acute appendicitis in children. AB - The aim of the study was to investigate the place of imaging in the diagnosis and treatment of acute appendicitis. The files of 2,427 children with suspected acute appendicitis were reviewed for clinical management and operative findings. The sample was divided into 3 groups at time of admission: (1) before diagnostic imaging was available in our department (1991-1994); (2) after the introduction of imaging studies on a random basis in equivocal cases (1995-1998); and (3) after a policy was formulated for ultrasound use in all equivocal cases followed by computed tomography if necessary (1999-2000). Results showed that the rate of misdiagnosis decreased from 13.2% in group 1 to 6.5% in group 2 and 6.1% in group 3. False-positive findings (normal appendix with positive scan) were noted in 16.7% of group 2 and 25% of group 3; false-negative findings (appendicitis at surgery with negative scan) in 23.8% and 9.5%, respectively. Computed tomography was performed in 8 children and prevented unnecessary surgery in 4 of them. We conclude that in equivocal cases of acute appendicitis, imaging studies performed by skilled operators can improve the accuracy of diagnosis, saving patients unnecessary surgery, and identifying other conditions that mimic appendicitis. PMID- 12635980 TI - Influence of breast-feeding on mental and psychomotor development. AB - The purpose of this study was to analyze the effects of breast-feeding on cognitive and motor development. A prospective cohort study was carried out enrolling 249 babies from 2 rural and urban areas. Cognitive development was assessed at the age of 18 months. Results show that adjusted scores on the mental development index were 4.6 points higher in babies breast-fed for longer than 4 months than in those breast-fed for less time. No differences in psychomotor development were found as a function of type of feeding. A statistically significant linear correlation was observed between the duration of breast feeding and scores for Bayley's mental development index. These data suggest that breast-feeding for longer than 4 months has a positive effect on the baby's mental development measured at 18 months of age. Parental intelligence quotient also seems to have an influence on cognitive development. PMID- 12635981 TI - Voiding dysfunction in pediatric patients. AB - The goals of this study were to describe the pattern of voiding disorders in children in our community, to describe clinical criteria for making the specific diagnoses, and to comment on management. The medical records of 226 children referred because of voiding dysfunction or urinary tract infections (UTI) were evaluated. Children with normal voiding patterns when uninfected, with monosymptomatic nocturnal enuresis, and with known neurologic or anatomic abnormalities were excluded. Detrusor instability, an abnormal voiding pattern characterized by urgency with or without frequency, was the diagnosis in 175 of the 226 children. Children with detrusor instability who used various posturing maneuvers to avoid urinary incontinence had a significantly higher incidence of UTIs than those who did not attempt to obstruct urine outflow. Detrusor instability appeared to be secondary to constipation in 19 of the children. The other diagnoses were extraordinary daytime urinary frequency, infrequent voiding, monosymptomatic daytime wetting, transient voiding dysfunction, giggle incontinence, dysfunctional voiding, and unexplained dysuria. It is concluded that children with detrusor instability who use posturing maneuvers to avoid incontinence are at high risk for recurrent UTIs. Constipation is 1 cause of detrusor instability. Dysfunctional voiding, the form of voiding dysfunction most likely to result in renal damage, was present in only 2 of 226 children seen for voiding disorders. PMID- 12635982 TI - Melatonin in children and adolescents with insomnia: a retrospective study. AB - Effectiveness and tolerability of melatonin was assessed in 32 children (mean age 9.6 +/- 4.5 years) with chronic sleep initiation and sleep maintenance problems treated naturalistically in a pediatric sleep medicine center. Children received melatonin for an average of 2.1 +/- 2.0 months at a final average dose of 2.0 +/- 1.2 mg administered 1 hour before bedtime. Twenty-nine (90.6%) children exhibited partial improvement to complete resolution of their sleep problems as measured by sleep latency time and number of awakenings reported by parents. Thus, melatonin may be effective, safe, and well tolerated in the treatment of chronic insomnia in children. PMID- 12635983 TI - Human growth hormone and gonadotropin releasing hormone analog combination therapy increases predicted height in short normal girls. AB - The "short normal" child constitutes a real challenge for the pediatric endocrinologist. In a subgroup of short normal children, puberty starts at a normal age but with low height, and hence, the final height is expected to be quite compromised. Efforts to improve the outcome in this group have been made in the past with equivocal results. We present the growth data of 8 short girls with normal growth hormone values on provocative testing and low height at puberty initiation. At intervention the height and the stage of puberty were 129.3 +/- 5 cm and II to III, respectively, and the predicted height was 148.8 +/- 2.6 cm. Gonadotropin releasing hormone analog, triptorelin (3.6 +/- 0.5 microg/kg/day) and growth hormone (0.5 IU/kg/week) were used in different sequential order and simultaneously in each child. The mean total treatment period was 47.6 +/- 11.2 months. The mean predicted and the mean final height in the total group were 148.8 +/- 2.6 and 154.5 +/- 3.6 cm, respectively (p:0.028). The final height did not differ from the target height (154.8 +/- 8 cm versus 154.5 +/- 3.6 cm), while in 4 children, the final height was greater than the target height. The height gain (delta Final height - Predicted height) was 5.7 +/- 1.3 cm. If we analyze separately the girls in whom growth hormone was started first and gonadotropin releasing hormone analog followed versus those who started the analog first, the delta Final height - Predicted height was 8 +/- 3 cm in the former and 4.8 +/- 3.1 cm in the latter (p:0.03). It seemed that the difference was accounted for by duration of growth hormone therapy (51.3 +/- 10.6 months versus 28.6 +/- 10.6 months) (p:0.026), rather than by other factors. In conclusion, under the conditions of the present study, the combination of puberty arrest and growth hormone therapy significantly improved predicted height. The most significant determinant of the height gain was the duration of growth hormone therapy. PMID- 12635985 TI - Oral corticosteroids in cat-scratch disease. PMID- 12635984 TI - Measures of cardiac repolarization and body position in infants. AB - Sudden Infant Death Syndrome (SIDS) is the most common cause of death in children between 1 and 6 months of age. Recent data suggest that a prolonged QTc interval on the 12-lead electrocardiogram (ECG) is associated with SIDS. Prone body position during sleep is also known to be a risk factor for SIDS; this has prompted the American Academy of Pediatrics to promote the "Back to Sleep" campaign. We postulated that the QTc interval in infants might change as a function of body position, linking the observations relating body position and QTc interval to SIDS. We recorded ECGs in a group of infants in both the supine and prone position to determine if the QTc interval and QT dispersion differ between the 2 positions. Forty-seven standard 12-lead EGGs and high-amplitude, rapid-sweep 12-lead EGGs were performed on 45 healthy infants (mean age 26 +/- 40 days) in both the supine and prone positions. The infants were asleep in a quiet, restful state. The ECGs were reviewed by 2 investigators blinded to the position of the infants during recording. Measurements included the average QTc interval (using Bazett's correction) and QT dispersion (the difference between the longest and the shortest QT intervals on a standard 12-lead EKG). The study was designed to detect a 3% difference in QTc interval with 80% power and alpha = 0.05. All subjects had telephone or clinical follow-up at 1 year. The average QTc interval was 403 +/- 20 milliseconds (msec) in the supine position and 405 +/- 27 msec in the prone position (p = NS). The QT dispersion was 20 +/- 12 msec in the supine position and 22 +/- 13 msec in the prone position (p = NS). One infant in the study group died of SIDS at the age of 3 months. The EGG of this patient revealed a QTc interval of 382 msec in the supine position and 407 msec in the prone position; the QT dispersion was 34 msec in the supine position and 34 msec in the prone position. We found no difference in QTc interval or QT dispersion as a function of body position in healthy infants resting quietly. Prolongation of the QTc interval is unlikely to explain the increased risk for SIDS associated with prone body position in the general population of healthy infants, unless patients with long QT syndrome are somehow more influenced by body position than normal patients are. PMID- 12635986 TI - Type 1 diabetes mellitus masked by primary adrenal insufficiency in a child with autoimmune polyglandular syndrome type 2. PMID- 12635987 TI - Family functioning in children with thalassemia. PMID- 12635988 TI - A clinical study in China of neonatal conjunctivitis caused by Chlamydia trachomatis. PMID- 12635990 TI - What are we without risk? The physician organization at a crossroads. AB - The managed care market in Chicago is experiencing rapid change. As health maintenance organization (HMO) enrollment flattens or even declines, and capitation becomes less sustainable for many, physician organizations are reevaluating their continued participation in risk-based contracts and are struggling to define their future roles. Physician organizations are looking for new ways to provide value to their physician members. Physician hospital organizations (PHOs) in particular are reassessing how the organization can continue to serve the interests of both the physicians and their hospital partners. To better understand the concerns of physician organizations, The Lowell Group surveyed Chicago area provider executives on their top issues. Three major concerns emerged: (1) protecting the financial health of the organization; (2) predicting the future of the managed care industry; and (3) evolving the physician organization to meet changing market conditions. Ultimately, physician organizations must make business decisions that support their true goals-serving patients and purchasers of care, physician members, and the organization's owners. PMID- 12635989 TI - An infant in septic shock. PMID- 12635991 TI - Costs of poor birth outcomes among privately insured. AB - Despite expansions in the public insurance coverage of pregnant women, concerns over poor birth outcomes remain. Poor birth outcomes occur among publicly and privately insured women, however, thereby imposing excess costs on employers and their insurers. Data from a large sample of privately insured for 1996 are used to examine these outcomes and costs. Almost one-fourth (24.3 percent) of the infants in our matched sample of 12,020 deliveries was premature or had other problems at birth. Costs for these infants accounted for 82 percent of the total $56 million spent on sample infants. The incremental cost of infants with poor birth outcomes versus those with normal, full-terms was approximately $14,600. We found that these relative costs had increased over time due perhaps to the increased technology and intensity of services used to save infant lives. We also found that factors other than maternal and infant complications affected cost variations. For example, employers located in the Northeast, hiring older mothers, and in unionized sectors have higher prenatal, delivery, and infant costs. PMID- 12635992 TI - Mergers and health care organizations. AB - The concerns over ever-rising health care costs has motivated physicians, hospitals, and insurance carriers to search for ways to increase efficiency, decrease costs, and maintain or improve quality. Mergers have been recognized as having the capability to accomplish all three goals-but not without some concerns. This article's intent is to provide the reader with basic knowledge that will assist in understanding the discussion regarding the appropriateness of mergers in health care. It reviews issues raised by mergers and examines a few case studies of merged health care organizations. PMID- 12635993 TI - The high price of quality: a cost analysis of NCQA accreditation. AB - Payers and consumers across the United States are expressing discontent with their health plans. We are witnessing a market-based demand for accreditation as a guarantee of acceptable quality in health services. Increasingly, accreditation by the National Committee for Quality Assurance (NCQA) is being viewed as the "gold standard" for health plans. This article presents a model for allocating resources and a cost analysis of anticipated expenditures associated with achieving this objective. PMID- 12635994 TI - The relationship between quality of care and financial performance in nursing homes. AB - Changes in the reimbursement structure of the Medicaid and Medicare programs have caused nursing homes to face severe revenue restraints. In the hopes of alleviating the effect of payment cutbacks on their financial performance, nursing homes have been instituting quality improvement initiatives. The goal of this study was to examine the relationships of quality of care with revenues, private-pay market share, and costs in the nursing home industry, and how these dynamics interplay to affect financial performance. This goal was achieved by using secondary data consisting of: (1) the Minimum Data Set Plus (MDS+); (2) the Health Care Information Analyst (HCIA) nursing home data set; and (3) the On-line Survey Certification of Automated Records (OSCAR) data set. Structural equation modeling (SEM) using maximum likelihood estimation was used to examine the total, direct, and indirect effects of the variables. Findings indicate that nursing homes that produce high quality care are able to achieve lower resident costs and in the process, report better financial performance than those facilities producing lower quality care. On the other hand, quality of care provided was not significantly associated with the revenues or private-pay market share of the nursing home. Overall, the total effects of quality to financial performance were positive (.055). PMID- 12635995 TI - Online benefits solutions--a new trend in managing employee benefits programs. AB - This article focuses on the array of online benefits solutions offered by technology companies and reports the benefits to both employers and employees. Some of the benefits include reduced paperwork, reduced errors, and reduced administration costs. Companies that can deliver these benefits will be in great demand to help manage benefits programs and streamline the administrative processes. PMID- 12635996 TI - Health financing reform in sub-Saharan Africa: major constraints, goals, and strategies. AB - Health financing reform became a critical element of public sector reform in sub Saharan Africa during the past decade. Within the framework of health sector reform, this article reviews the major constraints, goals, and strategies for health financing reform in sub-Saharan Africa. It identifies shrinking budgetary resources, increasing demand for health services, and rising health care costs as the primary factors driving the sub-region's health financing reform agenda. In light of these constraints, the article defines the major goals and the strategies for health care financing reform employed by many sub-Saharan African countries. PMID- 12635997 TI - Remodelling after surgical repair of atrial septal defects within the oval fossa. AB - In a retrospective study, we analysed the data from 101 adults with echocardiographic follow-up after surgical repair of defects within the oval fossa at a mean age of 35 +/- 17 years; 56% of the cohort being above the age of 30 years. Mean age at follow-up was 44 +/- 18 years, and length of follow-up was up to 40 years (11 +/- 12 years). At follow-up, atrial fibrillation or flutter was present in one quarter. Dilation of the right atrium, found in 64%, of the left atrium, found in 44%, and of the right ventricle, found in 29%, were also frequent, as well as pulmonary arterial hypertension, which was found in 30%. Diminished right ventricular ejection fraction, in contrast, was very rare, found only in 1%, and abnormal left ventricular ejection fraction was not encountered. By multivariate analysis, predictors for right or left atrial, or right ventricular, dilation were age at follow-up, degree of tricuspid regurgitation, pulmonary hypertension, and/or atrial fibrillation. In a subset of 21 patients in sinus rhythm, we correlated prospectively the diastolic and systolic function of both ventricles with levels of brain natriuretic peptide, comparing values to those of 20 age-matched controls with a mean age of 46 +/- 14 years. Levels of brain natriuretic peptide were significantly higher in patients than in controls (p = 0.006), and correlated significantly with diastolic dysfunction (p = 0.007) and left atrial size (p < 0.0001). In the long-term follow-up after surgical repair of defect within the oval fossa, therefore, complete normalization of heart size and function is rare. Despite preserved systolic function, persistent diastolic dysfunction is common and is associated with elevated levels of brain natriuretic peptide, which may explain the late occurrence of atrial arrhythmias. PMID- 12635998 TI - Low prevalence of femoral venous thrombosis after cardiac catheterizations in children: a prospective study. AB - OBJECTIVE: Cardiac catheterization is an important and frequent diagnostic intervention in children, but few systematic studies have explored the associated venous thrombotic complications. We have prospectively evaluated the prevalence of venous thrombosis, diagnosed by ultrasonography, in children catheterized at our department. MATERIALS AND METHODS: We examined 50 children with weight greater than seven kilograms for thrombosis of the femoral vessels. Prophylactic anticoagulation was given to half of the children who were to undergo left-sided catheterization. The enrolled children had ultrasonography of the site of puncture the day before, and the day after, the cardiac catheterization. During catheterization, blood samples were taken from 33 of the children for analysis of thrombophilic agents. RESULTS: The children, with a median age of 34 months, had been catheterized 103 times, comprising the 50 current and 53 previous procedures. Over the course of the current procedure, interventions of various types were performed in 18 children (36%). We diagnosed thrombophilic predispositions in four patients. Despite the thrombophilic tendencies, and a high frequency of interventions, we did not encounter a single case of femoral venous thrombosis. Nor did we find any evidence of arterial thrombosis. CONCLUSION: Our study indicates that catheterization procedures in children are currently associated with a low prevalence of femoral venous thrombosis. Continuous assessment of vascular complications, nonetheless, is still required to prevent late effects, and to ensure the best quality of cardiac catheterisations in children. PMID- 12635999 TI - Evidence for lack of myocardial injury in children with acute rheumatic carditis. AB - Despite pathologic evidence of myocardial inflammation, the significance of myocarditis in children with acute rheumatic carditis remains controversial. Elevations in cardiac troponin I have been demonstrated in other forms of myocarditis. The purpose of our study was to determine if levels of cardiac troponin I are elevated, suggesting myocardial injury, in patients with acute rheumatic carditis. We identified all those patients with acute rheumatic fever, presenting between July 1998 and December 2000, who had clinical evidence of carditis, such as a new murmur of mitral or aortic regurgitation, and who had an echocardiogram, measurements of levels of cardiac troponin I, erythrocyte sedimentation rate, and/or C-reactive protein performed at the time of presentation. Their charts were reviewed for demographic and clinical data. Echocardiograms were reviewed for severity of aortic and mitral regurgitation, and measurements made of left ventricular ejection fraction, fractional shortening, and end-diastolic dimension. We found 16 patients with acute rheumatic carditis, ranging in age from 2.0 to 16.1 years, with just over one third having symptoms of congestive heart failure. All patients had evidence of acute inflammation. There was a significant relationship between symptoms and severity of mitral regurgitation. No patient had elevated levels of cardiac troponin I level. The fact that levels of cardiac troponin I are not elevated in the serum of children with acute rheumatic carditis suggests that there is minimal myocytic necrosis in this setting. This supports the concept that acute valvar regurgitation is the major hemodynamic abnormality in these patients. PMID- 12636000 TI - Bony density in adolescents after surgical repair of tetralogy of fallot: a comparative study with healthy adolescents. AB - BACKGROUND: Adverse influences arising in fetal life or immediately after birth have a permanent effect on body structure, physiology and metabolism. Evidence is now accumulating that programming of bone growth might be an important contributor to the later risk of osteoporosis. Long-term morbidity and mortality associated with retralogy of Fallot is not completely known. The aim of the present study was to evaluate the state of the bones in adolescents after surgical repair of tetralogy of Fallot, so as to ascertain any possible repercussions of the disease on bone mineralization. MATERIAL AND METHODS: We studied 34 adolescents with repaired tetralogy of Fallot, between the ages of 11 and 18 years, to establish their nutritional status, in terms of height, weight, and skinfolds, their body composition using an anthropometric method, their sexual maturity according to Tanner, and their food-habits as based on 24-hour recall. Bone density was evaluated by lumbar dual-energy X-ray absorptiometry. We included 34 healthy eutrophic adolescents, matched for gender and age, as controls. RESULTS: No significant differences were observed between the patients and their controls concerning nutritional status, body composition, total energy intake and nutritional supply in macronutrients, calcium, phosphorus, magnesium and vitamin D. Bone mineral density, expressed in Z-score and g/cm2, was significantly higher in patients with tetralogy of Fallot (p < 0.01). The age at the time of the first surgical procedure, or at complete surgical repair, and the total number of surgical procedures, had no significant influence on nutritional status or bone mineralization. Gender, chronological age, sexual maturity and the index of body mass are the major determinants of bone density for both samples. Obese adolescents with repaired tetralogy of Fallot had a significantly higher bone density (p < 0.05) compared to undernourished or euthrophic patients. CONCLUSIONS: Being born with tetralogy of Fallot has no significant repercussion, by the stage of adolescence, on nutritional status, pubertal progression, and accretion of bone minerals subsequent to surgical repair. Nutritional status is the major influence on the accretion of bone mass. PMID- 12636001 TI - Nutritional support via percutaneous endoscopic gastrostomy in children with cardiac disease experiencing difficulties with feeding. AB - Adequate nutrition is crucial to the management of children and infants with cardiac disease. Difficulties with feeding are extremely common, and maintaining an adequate caloric intake, in order to achieve sustained growth, is often not possible without nutritional support. We retrospectively reviewed our experience between 1995 and 1999 in treating 37 children with cardiac disease who underwent percutaneous endoscopic construction of a gastrostomy to augment nutritional needs. We stratified the patients into those with cyanotic heart disease, when saturations of oxygen were less than 95%; those with non-cyanotic heart disease with saturations greater than 95%, and those with minor cardiac disease associated with a systemic disorder. Each group was compared to control children matched for age, sex, and diagnosis. We evaluated, the variation in standard deviation score for body weight over a median period of follow-up of 295 days. Improvements in the standard deviation score for body weight occurred in each of the groups, whereas children in the control groups demonstrated a decrease in standard deviation score for body weight. The median change of the score for body weight was significantly higher in patients managed with gastrostomy compared to controls. We conclude that supplementation using a gastrostomy tube allows the safe delivery of the caloric intake needed to support malnourished children with cardiac disease. PMID- 12636002 TI - Radiofrequency ablation of atrioventricular accessory pathways associated with discordant atrioventricular connections. AB - Discordant atrioventricular connections associated with Wolff-Parkinson-White syndrome increase the challenge of radiofrequency ablation. We report the results and techniques of radiofrequency ablation in three patients with discordant atrioventricular connections, including one patient having double outlet right ventricle with atrioventricular reentry tachycardias. There were two males and one female, aged 14 and 22 years old. We found four accessory pathways during our electrophysiological studies, with two of them manifest on the electrocardiogram, corresponding to left paraseptal and right midseptal regions. The electrophysiological study confirmed this localization, and showed two concealed accessory pathways in the right and left paraseptal regions. Radiofrequency ablation was successful in all cases without recurrence at a mean follow-up of 18.6 months. No complications were observed during the procedures. We conclude that radiofrequency ablation is feasible and effective in the ablation of accessory pathways in patients with discordant atrioventricular connections. PMID- 12636003 TI - Quality of life after repair of tetralogy of Fallot. AB - OBJECTIVE: To determine the quality of life in individuals with corrected tetralogy of Fallot. METHODS AND SUBJECTS: Questionnaires concerning quality of life were sent to all 87 surviving patients aged between 16 and 40 years who had undergone intracardiac repair of tetralogy of Fallot and follow-up in the Wessex Cardiothoracic Unit, and to 87 age and sex matched controls, with medically treated haemodynamically insignificant ventricular septal defects. RESULTS: The only significant difference found between the cases and controls was in requirements for schooling, where those with tetralogy of Fallot were more likely to require additional educational help at school (p = 0.044). For all other aspects of quality of life examined by the questionnaire, including social and genetic history, exercise ability, and health related quality of life, no significant differences were found. Different operative techniques, such as transjunctional patching, right ventriculotomy, and previous palliative shunting, did not affect the quality of life of our population with Tetralogy of Fallot, on average twenty years after their surgery, although the range of operative techniques was limited. Neither age at surgery, nor time since surgery, was correlated with measurements of quality of life. CONCLUSIONS: Those who have undergone surgical correction of tetralogy of Fallot have a normal quality of life, with few differences compared to controls. PMID- 12636004 TI - Aerobic capacity in adults with tetralogy of Fallot. AB - BACKGROUND: We investigated the aerobic capacity of 168 adult patients who had undergone successful surgical repair of retralogy of Fallot at the University of Toronto Congenital Cardiac Centre for Adults. METHODS: We compared values of peak uptake of oxygen, peak heart rate, forced vital capacity, and forced expiratory volume in 1 second to predicted values for their age groups. RESULTS: The patients who had undergone surgical repair of tetralogy of Fallot demonstrated an overall diminished peak uptake of oxygen, at 51%,and peak heart rate, at 79%, compared to predicted values. No difference in peak aerobic capacity was found according to the initial surgical strategy of palliation or repair. CONCLUSIONS: Adult patients who have undergone surgical repair of tetralogy of Fallot have lower peak uptake of oxygen, and peak heart rate, compared to predicted values. The reduction in the peak heart rate may affect their exercise capacity. The peak uptake of oxygen also decreased with increasing age at the time of testing, and the age at surgical repair. PMID- 12636005 TI - Images in congenital heart disease. Atrial septal defect within the oval fossa with enlarged coronary sinus: three-dimensional echocardiography. PMID- 12636006 TI - Images in congenital heart disease. Reversal of shunting in pulmonary hypertension after treatment with oral Sildenafil. PMID- 12636007 TI - The aorto-ventricular tunnels. PMID- 12636008 TI - Pulmonary hypertension due to spontaneous premature ductal constriction in fetal life: association with right bundle branch block. AB - We describe a case of fetal pulmonary hypertension and tricuspid regurgitation due to non pharmacologically induced ductal constriction observed at 36 weeks' gestational age. The hypertension resolved spontaneously soon after birth, with no functional consequences. Right bundle branch block is the only permanent anomaly, still being seen on the electrocardiogram at the age of 34 months. PMID- 12636009 TI - Giant fibroma of the right ventricle. AB - Cardiac fibromas are rare benign tumours of connective tissue that occur most frequently in children within the left ventricle. Spontaneous regression has not been observed, and surgical intervention is usually required. We have successfully treated a 1-year old girl with a giant fibroma of the right ventricle using the principles of the Batista procedure. The diagnosis was primarily made using transthoracic echocardiography. PMID- 12636010 TI - Innocent murmurs: the perception of the parents versus that of the child. AB - Innocent murmurs in childhood are common, and often they do not reflect cardiac disease. We have performed a prospective review, by means of a questionnaire, to clarify the parental perception of the innocent murmur identified in their child. Whenever possible, depending on age, we also sought the input of the child. Of a cohort of 63 infants and children with innocent murmurs, a response was obtained from 40 parents and 8 children to an initial questionnaire, with 26 parents replying to a second questionnaire a year later. No parent reported any symptoms in their children, or had imposed any restrictions. Of the parents, however three sets considered regular review was necessary. We conclude that the diagnosis of an innocent murmur by a paediatric cardiologist, aided by investigations including cross-sectional echocardiography, and an opportunity for explanation and reassurance, led to the child being treated normally. PMID- 12636011 TI - Successful transluminal angioplasty of renal arterial stenosis using the transcarotid approach. AB - We successfully performed percutaneous transluminal angioplasty to treat severe renovascular hypertension with left ventricular failure in a 5-month-old infant. Using the transcarotid approach, we dilated the stenotic left renal artery without any difficulties, using progressively larger balloons designed for dilation of coronary arteries. PMID- 12636012 TI - Continuous medical education. PMID- 12636013 TI - The timing of surgery for asymptomatic severe aortic or mitral regurgitation for children. PMID- 12636014 TI - Severe cutaneous reactions associated with the use of human immunodeficiency virus medications. AB - Patients infected with human immunodeficiency virus are highly susceptible to adverse dermatological reactions to specific medications. Severe cutaneous conditions such as Stevens-Johnson syndrome and toxic epidermal necrolysis are associated with high morbidity and, notably for toxic epidermal necrolysis, high mortality. Although overall mortality from human immunodeficiency virus has dramatically declined owing to highly active antiretroviral therapy, these antiretroviral regimens have been associated with a wide spectrum of severe cutaneous reactions. We reviewed case reports and clinical trials in the English literature on Medline (1966 to 2001) and Aidsline (1980 to 2000) to determine the prevalence of Stevens-Johnson syndrome and toxic epidermal necrolysis attributable to the current FDA approved antiretroviral medications. We identified a total of approximately 50 patients who had Stevens-Johnson syndrome and/or toxic epidermal necrolysis associated with the use of 5 antiretroviral medications: 2 nucleoside reverse transcriptase inhibitors, zidovudine (2 patients) and didanosine (1 patient); 1 non-nucleoside reverse transcriptase inhibitor, nevirapine (42 patients); and 2 protease inhibitors, indinavir (1 patient) and amprenavir (an unspecified number within the 1% of over 1400 patients experiencing severe life-threatening reactions). Of the reports that specified the onset time of adverse reaction after initiation of treatment, 86% (19/22) of patients experienced reactions within 4 weeks. Ten of the approximately 50 patients were diagnosed with Stevens-Johnson syndrome or toxic epidermal necrolysis, due to specific antiretroviral medication, or a combination of medications identified by either resolution upon withdrawal, consistent biopsy findings or a positive rechallenge. The remainder of the identified patients were reported in articles lacking data regarding drug administration, reaction history or other details. PMID- 12636015 TI - Cutaneous microdialysis of uric acid level in the dermis: modification of in vitro recovery. AB - Cutaneous microdialysis is a new tool for direct sampling and real-time monitoring in dermatological research. However, its use in general laboratory practice poses several problems, one of which is determination of the relative recovery of the target material. Uric acid, one of the major antioxidants in the skin, was analysed using cutaneous microdialysis in 11 healthy subjects. Two methods for in vitro recovery were adopted, one in which a standard solution of uric acid was used and another in which serum was used. Although differences between the two methods were found, it is suggested that establishing the in vitro recovery using serum might be a simple approach for microdialysis. PMID- 12636016 TI - Cutaneous squamoproliferative lesions in kidney transplant recipients: an investigation of specific Epstein-Barr virus expression. AB - In a previous report we demonstrated Epstein-Barr virus expression in cutaneous squamous cell carcinomas from heart transplant recipients. In a comparative study, skin lesions from renal allograft recipients were investigated for the presence of Epstein-Barr virus. Thirty cutaneous squamoproliferative lesions from 10 kidney transplant recipients were examined for Epstein-Barr virus-specific gene expression. The techniques used for detection were polymerase chain reaction, in situ hybridization and immunohistochemistry. Epstein-Barr virus DNA was not detected by polymerase chain reaction in the neoplasias, and only single Epstein-Barr virus-positive carcinoma cells were shown by in situ hybridization in three cases of infiltrative squamous cell carcinomas. Immunohistochemistry for Epstein-Barr virus latent membrane protein 1 showed a negative result in all samples. These findings differ from our earlier investigations of cutaneous squamous cell carcinomas from heart transplant recipients where Epstein-Barr virus expressions were common. This may indicate that the part Epstein-Barr virus plays in the development of post-transplant, cutaneous squamoproliferative disorders is related to type of organ transplantation and/or grade of immunosuppression. PMID- 12636017 TI - Atopy patch tests in young adult patients with atopic dermatitis and controls: dose-response relationship, objective reading, reproducibility and clinical interpretation. AB - The clinical interpretation and reproducibility of atopy patch tests was studied in 23 selected young adult patients with atopic dermatitis and 25 healthy controls using standard inhalant allergens. Non-invasive measurements were used for objective assessment of test reactions and the participants were retested after 6 weeks. Ten of 19 (53%) evaluable patients with atopic dermatitis had at least one positive atopy patch test. However, there was no clear clinical relevance of the atopy patch test results when related to patient history and distribution of dermatitis. Reproducible and dose-dependent results were obtained with Dermatophagoides pteronyssinus, grass and cat with a reproducibility rate of 0.69 to 0.81 in patients and 0.60-0.96 in controls. A unique finding was a significant positive correlation between a positive atopy patch test, allergen dose and increase in transepidermal water loss and erythema, while measurement of capacitance did not distinguish between positive and negative reactions. The results of the present study do not support the routine use of atopy patch tests in the evaluation of adult patients with atopic dermatitis. PMID- 12636018 TI - Autosomal recessive congenital ichthyosis in Sweden and Estonia: clinical, genetic and ultrastructural findings in eighty-three patients. AB - Congenital (non-bullous) ichthyosis is a rare group of keratinizing disorders which can be tentatively subclassified based on clinical criteria, analysis of transglutaminase 1 gene mutations and electron microscopy of epidermis. We studied 83 patients who were all on topical therapy and in 16 cases also on oral retinoids. Three main groups of patients were distinguished: (A) those with transglutaminase 1 gene mutations (n=44), (B) those without transglutaminase 1 gene mutations showing a coarse, generalized scaling (n=19), and (C) those without transglutaminase 1 gene mutations showing only fine or focal scaling (n=20). On clinical scoring, patients in group A were more hyperkeratotic and less erythematous than those in group B (p < 0.05). Anhidrosis was recorded in nearly all patients (> or = 80%), but ectropion and a collodion phenotype at birth were more common in group A versus other groups. Ultrastructurally, a high frequency of type I (Anton-Lamprecht's classification) was found in all three groups (37-63%), 20 cases of type II in group A and a few cases of types III and IV in groups B and C, respectively. In conclusion, transglutaminase 1 gene mutation is a major cause of congenital ichthyosis in Sweden and Estonia, and is often associated with severe scaling and ultrastructural type II in corneocytes. The transglutaminase-unrelated cases are more heterogeneous, probably reflecting a more varied aetiology. PMID- 12636019 TI - Chronic active EB virus infection complicated with IgG3 subclass deficiency: an adult case treated with intravenous immunoglobulin and IFN-alpha. AB - A 60-year-old man presented with recurrent genital and oral ulcers, necrotic papules on his face and scalp, spiking fever and indurated skin erythema on the trunk. A diagnosis of chronic active Epstein-Barr virus infection and IgG3 subclass deficiency was made, and he was supplemented by intravenous gammaglobulin injection. The spiking fever was resistant to treatment, but the addition of systemic interferon-alpha therapy was partially effective in treating his clinical symptoms, although the patient eventually died from pulmonary effusions and cardiac insufficiency. PMID- 12636020 TI - Congenital reticular ichthyosiform erythroderma--ichthyosis variegata: a case report and review of the literature. AB - We present a 32-year-old white patient with congenital reticular ichthyosiform erythroderma, also known as ichthyosis variegata. The patient had typical clinical features with areas of normal appearing skin surrounded by erythematous hyperkeratotic patches. The smaller "confetti-like" patches were mostly present on the trunk, while the larger areas forming a reticular pattern predominated on the extremities. Ultrastructural findings demonstrated perinuclear vacuolization, binuclear cells and filamentous deposits compatible with congenital reticular ichthyosiform erythroderma. Besides presenting the case, we review the literature on this rare disorder of keratinization. PMID- 12636021 TI - Congenital elephantiasis-like lymphangiomatosis of a lower limb. AB - The case of a newborn girl with a rare, giant, congenital, tissue lymphangioma giving rise to elephantiasis of the right lower limb is presented. The different imaging methods, especially magnetic resonance imaging, showed no extension of the lesions into the deep structures. At the age of 2 years, the child underwent a roentgenographic skeletal survey, which revealed osteolytic lesions in the femurs and the right tibia. There was no clinical evidence of systemic involvement. The place of this affection among the different lymphatic malformations was discussed and the diagnosis of elephantiasis-like lymphangiomatosis of the limb, an extremely rare disorder, has been retained. Early surgical reduction was performed, followed by application of a pressure dressing. Five years later the result remains satisfactory, but the excision of a persistent fluid-filled pouch around the knee will probably be necessary in the future. PMID- 12636022 TI - Orange-induced skin lesions in patients with atopic eczema: evidence for a non IgE-mediated mechanism. AB - Oranges are suspected of inducing adverse skin reactions in patients with atopic eczema. We studied 21 adult patients with atopic eczema and a history of adverse reactions to oranges and 10 patients without. A dietary history, skin tests, serum IgE and oral provocation tests with oranges were obtained. Severity of eczema was monitored by SCORAD, and serum tryptase, eosinophil cationic protein and urinary methylhistamine were measured. No allergic reactions were found to orange in skin prick or patch tests. However, 23 patients (74%) had specific serum IgE to orange. Oral provocation testing resulted in pruritic eczematous or maculopapular skin lesions predominantly at the predilection sites in 16 patients (52%). The SCORAD increased significantly in patients positive to the oral provocation test (p <0.05). Specific IgE to orange did not correlate with the clinical outcome of the oral provocation test. No significant changes were found in serum mast cell tryptase, eosinophil cationic protein or in urinary methylhistamine excretion. The negative results in the skin tests and a lack of correlation between specific IgE and oral provocation tests indicate that non-IgE mediated mechanisms are involved in cutaneous adverse reactions to oranges in patients with atopic eczema. PMID- 12636023 TI - Activation markers in severe atopic dermatitis following extracorporeal photochemotherapy. PMID- 12636024 TI - Bilateral nevus comedonicus: efficacy of topical tacalcitol ointment. PMID- 12636025 TI - Bullous congenital ichthyosiform erythroderma: safe and effective topical treatment with calcipotriol ointment in a child. PMID- 12636026 TI - Extensive striae distensae as a result of topical corticosteroid therapy in psoriasis vulgaris. PMID- 12636027 TI - Rifampicin-induced burning: an unusual presentation. PMID- 12636028 TI - Rosai-Dorfman disease presenting as large pigmented indurated plaques. PMID- 12636029 TI - Tinea incognito on the hand causing a facial dermatophytid reaction. PMID- 12636030 TI - Perineurioma: a tendon sheath fibroma-like variant in a distal subungual location. PMID- 12636031 TI - Areolar sebaceous hyperplasia. PMID- 12636033 TI - Facial Majocchi's granuloma caused by Trichophyton tonsurans in an immunocompetent patient. PMID- 12636032 TI - Mycosis fungoides restricted to the palms: efficacy of methotrexate. PMID- 12636034 TI - Detection of Bartonella henselae DNA using polymerase chain reaction assay in patient with cat scratch disease. PMID- 12636035 TI - Systemic contact dermatitis from ethylenediamine in an aminophylline preparation presenting as the baboon syndrome. PMID- 12636036 TI - Psoriasis guttata in association with hepatocellular carcinoma. PMID- 12636037 TI - Oral paraneoplastic pemphigus associated with renal malignancy. PMID- 12636038 TI - Bergeyella zoohelcum septicaemia of a patient suffering from severe skin infection. PMID- 12636039 TI - Acute generalized exanthematous pustulosis resembling toxic epidermal necrolysis caused by famotidine. PMID- 12636040 TI - Syngeneic acute graft-versus-host disease. PMID- 12636041 TI - Evidence-based laboratory medicine: addressing bias, generalisability and applicability in studies on diagnostic accuracy. The STARD initiative. PMID- 12636042 TI - Semen polymorphonuclear neutrophil leukocyte elastase as a diagnostic and prognostic marker of genital tract inflammation--a review. AB - Elastase is a protease released by polymorphonuclear neutrophils (PMN) during the inflammatory process. Since 1987, seminal elastase-inhibitor complex (Ela/alpha1 PI) has been proposed as a marker of male silent genital tract inflammation. Measured by immunoassay in seminal plasma, Ela/alpha1-PI at a cut-off level of > or = 230 microg/l, is useful in the detection of genital tract inflammation. The prevalence of increased seminal Ela/alpha1-PI in infertile men is significantly higher than that observed in fertile men. The Ela/alpha1-PI level is positively correlated with other seminal fluid markers of male genital tract inflammation: reduced semen volume, citric acid, fructose, and increased albumin, complement component C3, caeruloplasmin, immunoglobulins IgG and IgA, and cytokines interleukins-8 and -6. A higher seminal Ela/alpha1-PI level is significantly associated with tubal damage in female partners. After antibiotic therapy, a decrease of Ela/alpha1-PI level is observed. The presence of tubal damage in the partner may negatively affect the response to antibiotic treatment. A higher seminal Ela/alpha1-PI is associated with lower percentage of sperm with single stranded deoxyribonucleic acid (DNA) and better fertilization rate in in vitro fertilization. Besides infertility, the determination of Ela/alpha1-PI is useful to confirm the presence of prostate and other male accessory gland bacterial inflammation. Screening for PMN Ela/alpha1-PI is easy to perform and reproducible and is a reliable quantitative test for diagnosis and prognosis of silent genital tract inflammation of couples. Moreover, sequential determinations allow the follow-up of inflammation during and after therapy. PMID- 12636043 TI - Total oxyradical scavenging capacity toward different reactive oxygen species in seminal plasma and sperm cells. AB - The objective of this work was to evaluate the capability of both seminal plasma and sperm cells to scavenge different forms of oxyradicals and the possible correlation with sperm motility parameters. A total of 14 individuals were analyzed by computer-assisted sperm analysis (CASA) and the results integrated with the measurement of total oxyradical scavenging capacity (TOSC) toward peroxyl radicals, hydroxyl radicals and peroxynitrite in seminal plasma and spermatozoa. TOSC values revealed some significant correlation with kinetic sperm cell parameters, including curvilinear velocity (VCL), straight-line velocity (VSL) and linearity (LIN). A lower antioxidant capacity toward hydroxyl radical was found in the seminal fluid of men with reduced sperm motility. Such correlations were not found with peroxyl radicals and peroxynitrite, neither when TOSC values were analyzed in spermatozoa. The TOSC assay is a useful tool for studying the relationship between oxyradical toxicity and abnormal sperm cell motility. Although further investigations are needed, the data clearly establish different role for various forms of oxyradicals, i.e., hydroxyl radicals, in altering sperm motility. Measurement of TOSC is suggested as a useful means of indicating relationship between reactive oxygen species and sperm cell kinetics in clinical trials and antioxidant-based treatments. PMID- 12636044 TI - Identification of new polymorphisms in the CACNA1S gene. AB - We identified four novel polymorphisms in the CACNA1S gene that encodes the alpha1-subunit of the dihydropyridine receptor. Mutations in this gene are associated with two genetic diseases: malignant hyperthermia and hypokalemic periodic paralysis. The nucleotide substitutions c2403T --> C and c5398T --> C did not result in amino acid replacement, the nucleotide substitution c4475C --> A caused the replacement of the Ala1492 with an Asp residue and an A insertion was identified in intron 36. By using methods based on digestion with restriction enzymes we calculated the frequencies of these novel polymorphisms, as well as heterozygosity, in normal subjects from southern Italy. PMID- 12636045 TI - 3-Nitrotyrosine in atherosclerotic blood vessels. AB - The effect of peroxynitrite on the development of atherosclerosis is one of the major foci of recent studies. Here, the cytotoxic effect of peroxynitrite was investigated by quantitatively measuring nitrated tyrosine, 3-nitrotyrosine (3 NT) levels in atherosclerotic blood vessels. Atherosclerotic vessels were obtained from the patients who underwent either coronary artery or peripheric artery bypass surgery. Internal thoracic arteries of the patients were treated as non-atherosclerotic control vessels. 3-NT was measured by reverse-phase HPLC and plasma nitrite-nitrate levels were measured by spectrophotometry. 3-NT levels were significantly elevated in atherosclerotic vessels (46.6 +/- 23.3 nmol/mg protein, n = 15; p < 0.001) in comparison to control vessels (15.8 +/- 2.5 nmol/mg protein, n = 10). Vessel 3-NT correlated weakly with plasma nitrate levels (r = 0.38). Thus, atherosclerotic arteries have higher 3-NT levels than non-atherosclerotic blood vessels. PMID- 12636047 TI - Assessment of intestinal permeability: enzymatic determination of urinary mannitol, raffinose, sucrose and lactose on Hitachi analyzer. AB - The sugar absorption test is the usual test for measurement of intestinal permeability. After intestinal absorption of probe sugars the subsequently excreted sugars are measured in urine. We have developed four enzymatic methods for the measurement of the urinary concentration of the probe sugars mannitol, raffinose, lactose and sucrose. Mannitol, lactose and sucrose are directly measured on Hitachi 917 using mannitol dehydrogenase, beta-galactosidase and invertase, respectively, as enzyme reagents. Raffinose measurement needs a three hours preincubation with alpha-galactosidase, after which the liberated sucrose is measured. The analytical performances such as within- and between-run precision, linearity, lowest detection limit, interference of other sugars and comparison with a gas chromatographic method are described for the four methods. These methods are accurate an can easily be performed in any clinical laboratory. PMID- 12636046 TI - Molecular diagnosis of cystic fibrosis: comparison of four analytical procedures. AB - We compared the analytical accuracy, times, costs and the detection rate of four procedures for the molecular analysis of cystic fibrosis (CF). DNA from 127 subjects bearing different genotypes was tested by denaturating gradient gel electrophoresis followed by sequencing (reference procedure) and, for comparison, by ASO dot-blot (in-house), reverse dot-blot (Innogenetics), ARMS (Zeneca Diagnostics) and OLA-PCR (PE Applied Biosystem). To assess inter-observer variability, all samples were tested twice. To evaluate intra- and between-series variability, two samples were tested twice in each series. All the procedures yielded reproducible results and assigned the correct genotype to each sample. ASO dot-blot is the cheapest procedure but has the longest analytical time (> 24 h) and uses radioactive labeling. It can be adapted to analyze peculiar mutations in specific ethnic groups. ARMS from Zeneca Diagnostics is rapid (4 h), easy to perform, but, except for the AF508 mutation, does not distinguish the homozygote from the heterozygote genotype. It could be used for carrier analysis in families with known mutations. OLA-PCR has the highest detection rate in most ethnic groups, is automated for capillary electrophoresis but requires a high level of operator expertise: it is suitable when collected series of samples are analyzed from large patient cohorts. Reverse dot-blot is easy to perform and can be semiautomated: it can be used as first-line screening test. Given the heterogeneity of CF mutations, commercial kits should be developed to analyze mutations peculiar to specific ethnic groups. PMID- 12636048 TI - Effects of calcium carbonate, magnesium oxide and sodium citrate bicarbonate health supplements on the urinary risk factors for kidney stone formation. AB - We describe a model to illustrate different chemical interactions that can occur in urine following ingestion of individual and combined health supplements. Two types of interactions are defined: synergism and addition. The model was applied to eight healthy males who participated in a study to investigate the chemical interactions between calcium carbonate, magnesium oxide and sodium citrate bicarbonate health supplements on calcium oxalate urinary stone risk factors. Subjects ingested these components individually and in combination for 7 days. Twenty-four-hour urines were collected at baseline and during the final day of supplementation. These were analysed using standard laboratory techniques. Three different chemical interactions, all involving citrate, were identified: magnesium and citrate exerted a synergistic effect on lowering the relative superaturation (RS) of brushite; the same two components produced a synergistic effect on raising pH; finally, calcium and citrate exerted an additive effect on lowering the RS of uric acid. We propose that the novel approach described in this paper allows for the evaluation of individual, additive and synergistic interactions in the assessment of the efficacy of stone-risk reducing preparations. PMID- 12636049 TI - Glycerol metabolism and the determination of triglycerides--clinical, biochemical and molecular findings in six subjects. AB - Recent recommendations in the National Cholesterol Education Program Expert Panel on Detection, Evaluation and Treatment of High Blood Cholesterol in Adults (ATPIII) are expected to increase the number of triglyceride (TG) determinations and consequently the risk of misinterpretation of "non-blanked" results with co determination of free glycerol. Glycerol-kinase deficiency (GKD) is one cause of falsely elevated TG results. The natural history of isolated GKD with symptom free cases and cases with e.g. severe episodes of hypoglycemia and/or ketoacidosis challenges the laboratories to identify cases of GKD and family members at risk. "Blanked" methods reporting both glycerol and TG concentration are therefore desirable. Molecular studies of the glycerol kinase (GK) and DAX1 genes were performed on four cases of "persistent hypertriglyceridemia" found in an Italian population and on two pediatric cases with high serum glycerol concentration. Two new missense mutations were found (C358Y, T961). Molecular modeling on GK from E. coli, indicate that these mutations are located in parts of the enzyme important for enzyme formation or activity. One splice-site mutation, (IVS9A-1G>A), was found in two brothers. Splice-junction analysis indicates that it destroys the splice site and results in a mixture of mRNA. Deletion of the GK and DAX1 genes was found in one child with symptoms of adrenal failure. A female with glycerolemia and glyceroluria had normal GK activity but possibly slightly decreased ability to oxidize glycerol. PMID- 12636050 TI - Plasma lipid composition and LDL oxidation. AB - Low-density lipoprotein (LDL) oxidation in vivo depends on lipid composition and on plasma antioxidant status. The aim of our study was to investigate the relationship between plasma lipid composition and LDL oxidation and, in particular, to explore whether LDL-cholesterol/triglycerides ratio (LDL-C/TG) and LDL-cholesterol/high-density lipoprotein (HDL)-cholesterol ratio (LDL-C/HDL-C) can be used as predictive parameters of LDL oxidation in vivo. In 87 volunteers over a wide range of age plasma lipids and LDL oxidation were studied. Blood was collected after 12 h overnight fast. LDL oxidation was estimated by the level of conjugated diene (BDC) in the lipid fraction isolated from plasma after gradient ultra-centrifugation. The results were expressed as micromol/l (BDC/l) to evaluate the level of oxidized LDL, and as nmol of BDC for mg of LDL-cholesterol (BDC/LDL-C) for the evaluation of LDL oxidation degree. BDC/l correlated significantly with age, total and LDL-C, apolipoprotein B and TG, while BDC/LDL-C negatively correlated with total cholesterol, apolipoprotein B, LDL/TG and LDL/HDL ratios. Age of subjects significantly correlated with total and LDL-C and apolipoprotein B. TG have a significant inverse correlation with HDL-C. Our results support the hypothesis that among the several factors involved in LDL oxidation the most important determinants are LDL/TG. Plasma triglycerides appear to be very important even when circulating cholesterol levels are within normal limits. Moreover, we found that the LDL/HDL ratio is also very important with regard to the putative protective role of HDL against LDL oxidation in vivo. In conclusion, plasma lipid parameters must be evaluated not only for their absolute values but also for their mutual ratios as expression of plasma lipid homeostasis. Both LDL/TG and LDL/HDL ratios can be used as predictive parameters of in vivo LDL oxidation. PMID- 12636051 TI - Modified method to measure nasal potential difference. AB - Nasal potential difference (NPD) measurements have been proposed to assess defective ion transport in cystic fibrosis (CF). Implementing it routinely is, however, difficult. Therefore, a modified method based on nasal instillation in supine position at reduced flow rate was tested to evaluate its ability to discriminate CF from non-CF subjects. Classical and modified methods were compared in nine healthy subjects and there were no statistical differences. Following the new method, 97 tests were performed on 74 subjects divided in three cohorts: 21 CF patients and two control groups consisting of 19 patients with other pulmonary diseases and 34 healthy subjects. Twenty five children were enrolled in this study. Maximal NPD in CF patients (-44.9 +/- 2.5 mV) was significantly different from that obtained in control groups (-18.1 +/- 1.6 and 17.2 +/- 1.1 mV). Depolarization after amiloride also discriminated CF patients (25.9 +/- 1.4 mV) from control groups (10.5 +/- 0.9 and 8.1 +/- 0.7 mV). Marked repolarization following isoprenaline plus amiloride in low chloride solution was seen in control groups (-15.7 +/- 1.1 and -15.3 +/- 1.1 mV). We conclude that the modified method represents a simplified and equally effective approach to discriminate CF patients from non-CF subjects. Moreover, this method presents practical advantages for the patients related to hygiene and convenience, favoring its application in small children. PMID- 12636052 TI - Towards complete and accurate reporting of studies of diagnostic accuracy: the STARD initiative. AB - OBJECTIVE: To improve the accuracy and completeness of reporting of studies of diagnostic accuracy, to allow readers to assess the potential for bias in the study and to evaluate its generalisability. METHODS: The Standards for Reporting of Diagnostic Accuracy (STARD) steering committee searched the literature to identify publications on the appropriate conduct and reporting of diagnostic studies and extracted potential items into an extensive list. Researchers, editors, and members of professional organisations shortened this list during a two-day consensus meeting with the goal of developing a checklist and a generic flow diagram for studies of diagnostic accuracy. RESULTS: The search for published guidelines regarding diagnostic research yielded 33 previously published checklists, from which we extracted a list of 75 potential items. At the consensus meeting, participants shortened the list to 25 items, using evidence on bias whenever available. A prototypical flow diagram provides information about the method of patient recruitment, the order of test execution and the numbers of patients undergoing the test under evaluation, the reference standard or both. CONCLUSIONS: Evaluation of research depends on complete and accurate reporting. If medical journals adopt the checklist and the flow diagram, the quality of reporting of studies of diagnostic accuracy should improve to the advantage of clinicians, researchers, reviewers, journals, and the public. PMID- 12636053 TI - Novel and traditional biomarkers of bone turnover in postmenopausal women. AB - Urinary markers of bone collagen degradation such as N-terminal telopeptide, deoxypyridinoline, hydroxyproline and proline-hydroxyproline dipeptide as a novel marker were measured in both morning and daily urine samples collected from 40 postmenopausal women on 10 consecutive days. In addition, osteocalcin, bone alkaline phosphatase and procollagen C-terminal propeptide were analyzed in one serum sample from each of the women. DXA densitometry was employed to differentiate between osteopenic and osteoporotic women. Spearman rank correlation, factor analysis and receiver-operating characteristics were computed to evaluate the results. The highest correlation with a single common factor was found between telopeptide assayed in daily urine, followed by the dipeptide and hydroxyproline assayed in morning urine. These markers also showed high sensitivity for the diagnosis of osteoporosis, while osteocalcin had a high sensitivity for the diagnosis of bone disease. The concentrations of telopeptide, dipeptide and osteocalcin differed significantly between the osteopenic and osteoporotic subjects. Because of the ease of non-hydrolyzed urine processing, the speed of chromatographic analysis and the negligible costs, the dipeptide assay opens new possibilities in the field. PMID- 12636054 TI - Urinary plasma protein patterns in acute prostatitis. AB - We evaluated the diagnostic utility of urinary alpha1-microglobulin, alpha2 macroglobulin and albumin in the diagnosis of acute prostatitis. We studied 133 men (43 +/- 17 years) with, and a reference population (n=36, 41 +/- 16 years) without, urinary tract infection. Prostatectomy samples were used to study the potential interference between prostatic proteins and protein analysis. Urinary alpha2-macroglobulin/albumin ratio was significantly lower in prostatitis compared to the reference population, cystitis or acute pyelonephritis (p < 0.0001). Low alpha2-macroglobulin concentrations in prostatitis are due to inhibition (p = 0.0001) of the immune reaction between alpha2-macroglobulin in presence of polyclonal rabbit antibodies (used for immunonephelometry) by soluble prostatic proteins (+/- 60 kDa) which appear in urine in acute prostatitis. The urinary alpha1-microglobulin/creatinine ratio diagnoses acute pyelonephritis (sensitivity 100% and specificity 87%) and the urinary alpha2 macroglobulin/albumin ratio diagnoses acute prostatitis (sensitivity 100% and specificity of 90%). Stepwise multinomial logistic regression analysis reveals that urinary alpha1-microglobulin, alpha2-macroglobulin, albumin and creatinine provide optimal differentiation between acute pyelonephritis and acute prostatitis (pseudo R2=0.83; Loglikelihood -30.55, p < 0.000001). In conclusion, the combination of hematuria and absence of urinary alpha-2-macroglobulin is diagnostic for acute prostatitis. Even without hematuria, alpha2-macroglobulin remains lower compared to patients without prostatitis. PMID- 12636055 TI - Adequate sampling in cryoglobulinaemia: recommended warmly. AB - Propersample handling is important for the accurate and prompt diagnosis of cryoglobulinaemia as well as for obtaining correct results of other laboratory tests. In this case report, we present two patients for whom improper sampling delayed the diagnosis of cryoglobulinaemia. The routine sampling procedure was not sufficient for detecting these cryoglobulins and we demonstrated that the temperature dropped below 37 degrees C at different steps during the sample preparation. Adjusting the preanalytical procedure revealed that both patients possessed a cryoglobulin with high thermal insolubility. We conclude that in cryoglobulinaemia strict adherence to guidelines to keep the temperature >37 degrees C is crucial for sample collection and must be strongly recommended. PMID- 12636056 TI - A new modification of the Chiron ACS assay for total prostate-specific antigen achieves equimolar response characteristics and improves the detection of prostate cancer. AB - Nonequimolar-response assays for prostate-specific antigen (PSA) are criticized for overestimating total PSA in some men without prostate cancer (PCA), and underestimating total PSA in some men with PCA. We recently studied three nonequimolar-response PSA assays that had undergone modifications. While two of the studied assays achieved equimolar-response characteristics with improved areas under receiver operating characteristic (ROC) curves (AUC), the modification of the Chiron ACS PSA assay (ACS PSA2, Chiron) failed to achieve this. Recently, the ACS assay underwent another modification (ACS PSA, Bayer), which we investigated. Sera from 305 men (155 without and 150 with PCA, PSA > or = 2 and < or = 30 microg/l, Tandem-E) were measured using both modifications of the ACS assay and equimolar-response reference methods (Tandem-R free and Tandem E, Hybritech). Molar response relative to the reference method and clinical performance (comparison of AUCs) between the previous and new ACS assay modifications were studied. The new modification of the ACS assay (ACS PSA, Bayer) achieved equimolar-response characteristics but reported lower values (average 10%) than the Tandem-E assay. Compared to the previous modification (ACS PSA2, Chiron), a 3% improvement in AUC (p = 0.01) was found. Using results of the redesigned equimolar-response assay (ACS PSA, Bayer), we calculated that 6 of 155 men without PCA in this sample set could be spared unnecessary biopsy compared with the previous nonequimolar-response assay (ACS PSA2, Chiron) without missing additional PCA (90% sensitivity). These data provide additional evidence for clinical advantages of equimolar-response over nonequimolar-response PSA assay formats. PMID- 12636059 TI - On-line monitoring equipment for wastewater treatment processes: state of the art. AB - A (non-exhaustive) survey of new and existing technologies for the monitoring of wastewater treatment plants is presented. Emphasis is given to the way these sensors can provide insight in the ongoing (bio-) processes. Three different uses for sensors can be found: for monitoring (operator support), in automatic control systems and as tools for plant auditing/optimization/modelling by consultants. From this, sensors have been classified in two basic types: (i) reliable, simple and low maintenance sensors for day-to-day monitoring and control and (ii) advanced, higher maintenance sensors that are used in auditing, model calibration and optimisation. The paper is organized according to the typical unit processes of biological wastewater treatment systems: anaerobic digestion, activated sludge, nutrient removal and sedimentation. Attention is drawn to a number of practical problems associated with the use of sophisticated sensors in the harsh (dirty) conditions of wastewater treatment processes. The use of autocalibration and built-in sensor checks, cleaning systems and reliable sample preparation units is illustrated. The paper ends with a discussion of the applicability of the different sensors. PMID- 12636057 TI - Technical performance and diagnostic utility of the new Elecsys neuron-specific enolase enzyme immunoassay. AB - This international multicenter study was designed to evaluate the technical performance of the new double-monoclonal, single-step Elecsys neuron-specific enolase (NSE) enzyme immunoassay (EIA) and to assess its utility as a sensitive and specific test for the diagnosis of small-cell lung cancer (SCLC). Intra- and interassay coefficients of variation, determined in five control or serum specimens in six laboratories, ranged from 0.7 to 5.3 (inter-laboratory median: 1.3%) and from 1.3 to 8.5 (inter-laboratory median: 3.4%), respectively. Laboratory-to-laboratory comparability was excellent with respect to recovery and inter-assay coefficients of variation. The test was linear between 0.0 and 320 ng/ml (highest measured concentration). There was a significant correlation between NSE concentrations measured using the Elecsys NSE and the established Cobas Core NSE EIA II in all subjects (n = 723) and in patients with lung cancer (n = 333). However, NSE concentrations were systematically lower (approximately 9%) with the Elecsys NSE than with the comparison test. Based on a specificity of 95% in comparison with the group suffering from benign lung diseases (n = 183), the cut-off value for the discrimination between malignant and benign conditions was set at 21.6 ng/ml. NSE was raised in 73.4% of SCLC patients (n = 188) and was significantly higher (p < 0.01) in extensive (87.8%) as opposed to limited disease (56.7%). NSE was also elevated in 16.0% of the cases with non-small cell lung cancer (NSCLC, n = 374). It is concluded that the Elecsys NSE EIA is a reliable and accurate diagnostic procedure for the measurement of NSE in serum samples. The special merits of this new assay are the wide measuring range (according to manufacturer's declaration up to 370 ng/ml) and a short incubation time of 18 min. PMID- 12636058 TI - Measurements of complement factor H-related protein (BTA-TRAK assay) and nuclear matrix protein (NMP22 assay)--useful diagnostic tools in the diagnosis of urinary bladder cancer? AB - Between 1997 and 2000 we investigated in a prospective study the voided urine samples of all consecutive patients undergoing cystoscopy independent from their clinical background (n = 705) with the BTA-TRAK assay (Bard Diagnostics, Redmont, USA) detecting a complement factor H-related protein (CFHrP) and the NMP22 assay (Matritech, Newton, USA) measuring a nuclear matrix protein, which is supposed to be specific for bladder cancer. The individuals were divided into three groups concerning the clinical background: 233 patients had urological diseases, 268 patients had urinary bladder cancer and 150 patients had other urological malignancies. Based on the clinical findings we compared our results with well established diagnostic methods for urinary bladder cancer such as cytology and the detection of hematuria. In addition, we investigated urine samples from 30 healthy individuals and 24 patients with urinary tract infection without performing cystoscopy. Following the recommendations of the European Group on Tumor Markers we used 95% specificity for benign urological diseases and urinary tract infections, which resulted in a sensitivity of 17% for active bladder cancer for the BTA-TRAK assay and 31% for NMP22. We compared these results with the detection of hematuria (specificity: 72%) and cytology, which had a sensitivity of 64% and 89%, respectively. Subsequently, we calculated sensitivity and specificity for the detection of relapse of the disease. Again using 95% specificity, in this case for patients with no evidence of disease (NED), in patients with recurrent disease the BTA-TRAK assay showed 8% sensitivity as compared to 12% for the NMP22 assay. Due to an insufficient specificity and sensitivity, both tests can neither be clinically useful in screening of high risk patients, nor in primary diagnosis of bladder cancer. They cannot replace neither cystoscopy nor cytology. In the follow-up care more investigations may be necessary to prove the benefit of existing diagnostic strategies for the discrimination between active and inactive bladder cancer. PMID- 12636060 TI - Infrared optical sensors for water quality monitoring. AB - In-situ monitoring of water quality with particular emphasis on organic pollutants is a global priority topic in water analysis. Recent developments in optical sensor technology provide advanced analytical tools for continuous assessment of pollution levels in the liquid phase and in the gas phase. Infrared sensing schemes are among the most promising concepts due to inherent molecular specificity provided by absorption patterns of fundamental molecular vibrations of organic molecules. The advent of mid-infrared transparent optical fibers and waveguides, appropriate light source technology, such as quantum cascade lasers, and the potential for the development of highly integrated analytical devices based on microfabrication technology substantiates the trend towards spectroscopic sensing techniques. Chemical modification of the waveguide surface leads to enhanced analyte recognition based on tunable properties of enrichment or (bio)chemical recognition layers. Discussion of fundamental sensing technology is complemented by recent examples, highlighting the state-of-the-art in this dynamic research field. PMID- 12636061 TI - Transforming data into information. AB - In spite of a long history of automated instruments being deployed in the water industry, only recently has the difficulty of extracting timely insights from high-grade, high-volume data sets become an important problem. Put simply, it is now relatively easy to be "data-rich", much less easy to become information rich". Whether the availability of so many data arises from "technological push" or the "demand pull" of practical problem solving is not the subject of discussion. The paper focuses instead on two issues: first, an outline of a methodological framework, based largely on the algorithms of (on-line) recursive estimation and involving a sequence of transformations to which the data can be subjected; and second, presentation and discussion of the results of applying these transformations in a case study of a biological system of wastewater treatment. The principal conclusion is that the difficulty of transforming data into information may lie not so much in coping with the high sampling intensity enabled by automated monitoring networks, but in coming to terms with the complexity of the higher-order, multi-variable character of the data sets, i.e., in interpreting the interactions among many contemporaneously measured quantities. PMID- 12636063 TI - A multivariate calibration procedure for UV/VIS spectrometric quantification of organic matter and nitrate in wastewater. AB - A submersible UV/VIS spectrometer for in-situ real-time measurements is presented. It utilises the UV/VIS range (200-750 nm) for simultaneous measurement of COD, filtered COD, TSS and nitrate with just a single instrument. A global calibration is provided that is valid for typical municipal wastewater compositions. Usually high correlation coefficients can be achieved using this standard setting. By running a local calibration improvements concerning trueness, precision and long term stability of the results can be achieved. The calibration model is built by means of PLS, various validation procedures and outlier tests to reach both high correlation quality and robustness. This paper describes the UV/VIS spectrometer and the calibration procedure. PMID- 12636062 TI - Use of real-time data in environmental monitoring: current practices. AB - Water quality monitoring in Europe, especially in transboundary water courses has made steady progress during the last decades through establishment of international commissions. The main activities of these commissions include protection and management of the catchment, sustainable use of the river and establishment of Accident Emergency Warning Systems (AEWS). The latter could be effectively accomplished only through real-time monitoring. Concurrently real time data have been found important for the monitoring of potable water intake points, wastewater treatment plants, estuaries and in aquaculture. With the recognition of the diversified demand, there are a number of questions to be answered such as: (1) are we satisfied with the existing monitoring systems? (2) is standardisation of the measuring instruments a necessity? (3) do we have foolproof systems for data capture and transmission? (4) are there adequate procedures to analyse vast amount of data generated? We have to answer these questions urgently as the demand for real-time monitoring has been drastically increased. PMID- 12636064 TI - Influence of changes of the wastewater composition on the applicability of UV absorption measurements at combined sewer overflows. AB - The applicability of UV-absorption measurements at combined sewer overflows was investigated under different storm water conditions. Three specific rainfall events of different intensity, different duration and different intervals between the events were investigated. An individual evaluation of each single event resulted in an acceptable correlation between the UV absorption and the pollution measured as COD. A combined evaluation of the different events exhibited a very poor correlation which could not be used as pollution control parameter. Since the calibration procedure of the spectrometer used was not flexible enough to distinguish between different wastewater matrices from the different storm events the applicability of this type of spectrometer for the control of storm water overflows is questionable. PMID- 12636065 TI - Non-contact measurement of wastewater polluting load--the Loadmon project. AB - A new approach to the measurement of polluting load in crude wastewaters is described. The Loadmon project has developed an optical technique which enables suspended solids and organic load of a crude wastewater to be measured in an open channel. There is no requirement for any sampling equipment or sensor contact of any kind. The paper covers the laboratory study of wastewater optical properties,the prototype development, testing in the laboratory and field trials. Future plans for product development are outlined. PMID- 12636066 TI - Determination of sewer roughness and sediment properties using acoustic techniques. AB - Measurements have been carried out to determine the effect of porous sediments in a pipe on the attenuation and spectrum of the airborne acoustic field. The results show that the presence of even a relatively thin sandy layer results in a considerable increase in the acoustic attenuation over the broad frequency range. The measured value of the relative attenuation is in the range of 0.8 dB/m. The effect of the sediment on the acoustic spectrum is the reduction in energy of the propagating modes, which is an easily detectable phenomenon. These results pave the way for the development of the instrumentation for non-invasive characterisation of the parameters of sediments in wastewater systems. PMID- 12636067 TI - Accounting for sensor calibration, data validation, measurement and sampling uncertainties in monitoring urban drainage systems. AB - Assessing the functioning and the performance of urban drainage systems on both rainfall event and yearly time scales is usually based on online measurements of flow rates and on samples of influent effluent for some rainfall events per year. In order to draw pertinent scientific and operational conclusions from the measurement results, it is absolutely necessary to use appropriate methods and techniques in order to i) calibrate sensors and analytical methods, ii) validate raw data, iii) evaluate measurement uncertainties, iv) evaluate the number of rainfall events to sample per year in order to determine performance indicator with a given uncertainty. Based an previous work, the paper gives a synthetic review of required and techniques, and illustrates their application to storage and settling tanks. Experiments show that, controlled and careful experimental conditions, relative uncertainties are about 20% for flow rates in sewer pipes, 6 10% for volumes, 25-35% for TSS concentrations and loads, and 18-276% for TSS removal rates. In order to evaluate the annual pollutant interception efficiency of storage and settling tanks with a given uncertainty, efforts should first be devoted to decrease the sampling uncertainty by increasing the number of sampled events. PMID- 12636068 TI - Progress in sensor technology--progress in process control? Part 1: sensor property investigation and classification. AB - To ensure correctly operating control systems, the measurement and control equipment in WWTPs must be mutually consistent. The dynamic simulation of activated sludge systems could offer a suitable tool for designing and optimising control strategies. Ideal or simplified sensor models represent a limiting factor for comparability with field applications. More realistic sensor models are therefore required. Two groups of sensor models are proposed on the basis of field and laboratory tests: one for specific sensors and another for a classification of sensor types to be used with the COST simulation benchmark environment. This should lead to a more realistic test environment and allow control engineers to define the requirements of the measuring equipment as a function of the selected strategy. PMID- 12636069 TI - Progress in sensor technology--progress in process control? Part II: results from a simulation benchmark study. AB - To show the impact of sensor behaviour on the control result, four strategies for aeration control are tested using different sensor characteristics. It is demonstrated, on the one hand, how an increasing response time will limit the achievable control quality and, on the other hand, how a given sensor characteristic can be taken into account for the controller design. The presented tests show that an improvement potential by control for WWTPs is available but this potential is limited compared to proper DO control with fixed set-points. To activate this control potential, sufficient control authority must be available and a careful control design is required. It can be shown that using feedback control, sensors with a small response time have significant advantages compared to conventional sensors. Using feed forward control, the improvement potential by control is considerably higher and additionally, the sensor delay can be integrated into the controller design. The presented discussion is based on simulation studies performed on a standardised benchmark case. For these tests it was necessary to include sensor models into the simulation model. It can be stated that the usage of sensor models is necessary for the application of dynamic simulation for the design and evaluation of WWTP control and in general to achieve realistic results. PMID- 12636070 TI - Online sensing of volatile organic compounds in groundwater using mid-infrared fibre optic evanescent wave spectroscopy: a pilot scale test. AB - A prototype sensing system for in-situ monitoring of volatile organic compounds in contaminated groundwater was tested at a pilot scale plant. The sensor consists of a commercially available Fourier transform infrared spectrometer, connected to a 6 m long infrared transparent silver halide fibre optic cable. A 10 cm long core-only section at the centre of the fibre is mounted on a sensor head and coated with a hydrophobic polymer layer, while the remaining fibre is protected by Teflon tubing and thus not in contact with the surrounding media. The sensor head was immersed into the monitoring wells of the pilot plant testing the sensor system under circumstances close to field conditions and typical for in-situ measurements. The pilot plant consists of a 1 m3 cubic tank filled with gravel. A pump is used to circulate water horizontally through the tank, simulating a natural aquifer. The evolution of the concentration of analytes injected into the system is monitored with time using the developed prototype sensing system. The results are validated by corresponding sampling and analysis with headspace gas chromatography. PMID- 12636071 TI - Microelectrode array sensor for water quality monitoring. AB - A versatile microelectrode array sensor for water quality monitoring has been developed. The array fabrication, based on batch microelectronic processes, results in a highly stable passivation of the silicon chip surface and provides the possibility to use a backside contact. Packaging was optimized for on-line water operation at high pressures. Examples of applications include chlorine monitoring in drinking water, ozone monitoring in deionized water, dissolved oxygen in activated sludge and preliminary measurements of trace arsenic. PMID- 12636072 TI - New chalcogenide glass chemical sensors for S2- and dissolved H2S monitoring. AB - A non-optimised treatment of wastewaters containing organic and biological substances is very often accompanied by an accidental emanation of hydrogen sulphide H2S and therefore leads (i) to an undesirable odour in the vicinity of water treatment plants, and (ii) to a potential hazard for the neighbouring population. Fast, sensitive and reliable monitoring devices hence of significant importance. Chalogenide and chalcohalide glass are new promising membrane material for detection of heavy metal ions and toxic anions and particularly well adapted for continuous in situ monitoring and industrial process control. In the present paper, we will discuss analytical characteristics of new chalcogenide glass chemical sensors for detection of S2- and dissolved H2S, which slow reliable process control to be carried out at natural pH of wastewaters. PMID- 12636073 TI - An integrated sensor for the monitoring of aerobic and anoxic activated sludge activities in biological nitrogen removal plants. AB - An integrated sensor is developed as a tool for monitoring the activated sludge activity on which the performance of the treatment plant depends. The sensor provides information-rich data of high frequency obtained from respirometric titrimetric and nitrate measurements in one single set-up. The sensor is shown to successfully monitor and provide in-depth insight into nitrification, denitrification and carbon source degradation processes occurring in BNR plants. Based on the experimental results it is hypothesized the ratio of NUR to OUR rather reflects the rate of carbon source uptake (storage) under anoxic and aerobic conditions than growth process. PMID- 12636074 TI - Rapid automated detection of nitrification kinetics using respirometry. AB - There is no doubt that respirometry is a useful measurement principle in the field of wastewater treatment. Although a large variety of methods and case studies have been published, respirometry has become neither a standard tool for control nor for assessment and optimisation of treatment plants. drawback of the conventional method for determining nitrification kinetics is the long experimental time. This disadvantage can be avoided by "turning over" the experiment. Starting with low ammonia concentrations the steep slope of the Monod curve is measured first. The low concentration branch of the Monod curve is also the part where e.g. inhibition can be detected. Therefore the proposed procedure allows us to speed up the measurement of nitrification kinetics and to measure nitrification inhibition on-line. PMID- 12636075 TI - Automation in sludge dewatering by novel on-line characterisation of flocculation. AB - A novel on-line method and system for characterisation of sludge flocs in view of improving sludge dewatering has been developed. The characterisation of sludge flocs was carried out after the conditioning or the flocculation process. The system uses a conventional CCD-line scan camera providing an on-line monitoring of the relative floc size distribution by image processing procedures. The image processing procedure has been re-evaluated and adapted to the practical dewatering results, obtained from a chamber filter press of 250 x 250 mm size. A good correlation between the calculated sensor signal and the sludge dewaterability of digested sludge in terms of the up-concentration factor was found. Although different sludge compositions and flocculation systems have been tested within the whole experimental period of six months, a good reproducibility of this correlation was also found. A well balanced floc size distribution is necessary showing that not too many but still some fine flocs and also not too large but compact flocs lead to improved dewaterability. This has been illustrated by an extended range of floc size measurements ranging between 50 microm and 29 mm. The conditioning monitoring system would be suitable for the control of production of good size-balanced flocs to compensate fluctuations in sludge characteristics of the sludge to be conditioned. PMID- 12636076 TI - A software monitor for intermittent bacteria contamination in urban rivers. AB - Norwegian receiving waters are of such high water quality that authorities consider opening them for bathing. The leading parameter to monitor the quality of bathing waters is fecal coliform bacteria (FC). For this parameter no rapid detection method is available. The main objective of this case study was to find a way to quickly predict bacteria contamination by observing different online parameters such as flow, conductivity or spectral absorption coefficient (SAC). In this study historical data from 1994 to 2000 was analyzed, and over a period of five weeks water samples were taken and analyzed for bacteria. The analysis of the historical data revealed fundamental sampling problems, which made the data useless for the purpose of this study. The analysis of the data collected for this study showed that exceeding the bathing water standard for bacteria can be predicted by evaluating the SAC with an acceptable accuracy. Furthermore a simple river quality model was implemented, including bacteria as a load fraction. With the help of rain data and discharge predictions expected bacteria numbers exceeding the bathing water standard could also be forecast. PMID- 12636077 TI - Multivariate on-line monitoring: challenges and solutions for modern wastewater treatment operation. AB - In this paper, a number of challenges, which need to be overcome if multivariate monitoring of wastewater treatment operation is to be successful, are presented. For each challenge, one or several solutions are discussed. The methodologies are illustrated using an example from full-scale wastewater treatment operation. Some guidelines regarding choices of methods and implementation aspects are given. PMID- 12636078 TI - Novel monitoring concepts to acquire new water quality knowledge. AB - Two novel water quality monitoring concepts were developed: the HPLC-fingerprint for the monitoring of yet unidentified pollutants and the HPLC-Toxprint for the recognition of (unknown) toxic or genotoxic compounds. The paper describes applications of both concepts. The HPLC-fingerprint is used for the evaluation of the overall water quality in addition to the monitoring of individual pollutants. Based on their occurrence (frequency, concentration, location) a listing of unknown priority pollutants is set up. Participating waterworks monitor for these compounds using a dedicated HPLC-DAD library that contains the required compound data (UV-absorbance spectrum, retention time index). In five years of experience with this concept, the HPLC-fingerprint was also found very suitable for the retrieval of new priority pollutants in existing HPLC-fingerprint databases, providing historic data on these new compounds. The HPLC-fingerprint concept was also used as an Early Warning System for accidental spills or sabotage. The HPLC Toxprint was successfully applied in identifying genotoxicity (in the umu-test) in various waste water samples. By the application of LC-MS/MS genotoxicity could be assigned to acridine-derivatives in one of these wastewaters. To enable the evaluation of drinking water resources, the sensitivity of the HPLC-Toxprint was improved, now allowing the detection of pollutants with a 10% genotoxic potential as compared to 2-aminoanthracene (the positive control compound) at concentrations as low as 0.1 microg/l. PMID- 12636079 TI - Hydrological information system based on on-line monitoring--from strategy to implementation in the Brantas River Basin, East Java, Indonesia. AB - The worsening environmental situation of the Brantas River, East Java, is addressed by a comprehensive basin management strategy which relies on accurate water quantity and quality data retrieved from a newly installed online monitoring network. Integrated into a Hydrological Information System, the continuously measured indicative parameters allow early warning, control and polluter identification. Additionally, long-term analyses have been initiated for improving modelling applications like flood forecasting, water resource management and pollutant propagation. Preliminary results illustrate the efficiency of the installed system. PMID- 12636080 TI - Remote water quality monitoring "on-line" using LEO satellites. AB - Low Earth Orbit Satellites (LEOS) provide world-wide "nomadic" data-communication using low-cost and low-power "cellular like" equipment. LEOS allow us to also exploit those regions, which up to now were inaccessible by land-based wireless or geostationary (GEO) satellite systems. First, this paper compares satellite systems and summarizes features of the ORBCOMM LEO-system used. In the main part, a detailed report is given on a hydrogeological prototype implementation for remote water-quality monitoring: setup, obtained results, conclusions and further aspects as well as cost comparisons. As a result, LEO satellite data transmission may be a reliable low-cost alternative even to GSM-SMS for low data volume applications. PMID- 12636081 TI - On-line measurement data of wastewater systems via WAP mobile phones. AB - In order to support the operation of wastewater systems a system was developed which allows us to access plant data by standard mobile devices such as WAP mobile phones. This system is suited complement the standard application of alarm and message systems based for example on SMS or pager services. This technology provides useful options for mobile remote monitoring and remote control of automated plants. This technology is particularly appropriate for the use in remote facilities where no staff is available. The technology has been implemented succesfully and shows how standard IT and Internet technologies can be utilised to support the operation of spatially distributed plants with reasonable effort. Two implementations are presented which access plant data via WAP mobile phones and via mobile pocket PCs. First application experiences are presented. PMID- 12636082 TI - Marine natural products. AB - This review covers the literature published in 2001 for marine natural products, with 497 citations (373 for the period January to December 2001) and includes 793 compounds isolated from marine microorganisms and phytoplankton, green algae, brown algae, red algae, sponges, coelenterates, bryozoans, molluscs, tunicates and echinoderms. The emphasis is on new compounds and new stereochemical assignments (683 for 2001), together with relevant biological activities, source organisms and country of origin. Syntheses that confirm or revise structures or stereochemistries have been included (95), including any first total synthesis of a marine natural product. PMID- 12636083 TI - Twenty years of gibberellin research. AB - This review covers research into the chemistry and biology of the gibberellin family of plant bioregulators carried out in the author's laboratory over the past 20 years and has 231 references. PMID- 12636084 TI - Diterpenoids. AB - The review covers the isolation and structures of diterpenoids including labdanes, clerodanes, pimaranes, abietanes, kauranes, gibberellins, cembranolides, taxanes and marine diterpenoids. The literature from January to December 2001 is reviewed and 140 references are cited. PMID- 12636085 TI - The chalcone synthase superfamily of type III polyketide synthases. AB - This review covers the functionally diverse type III polyketide synthase (PKS) superfamily of plant and bacterial biosynthetic enzymes. from the discovery of chalcone synthase (CHS) in the 1970s through the end of 2001. A broader perspective is achieved by a comparison of these CHS-like enzymes to mechanistically and evolutionarily related families of enzymes, including the type I and type II PKSs, as well as the thiolases and beta-ketoacyl synthases of fatty acid metabolism. As CHS is both the most frequently occurring and best studied type III PKS, this enzyme's structure and mechanism is examined in detail. The in vivo functions and biological activities of several classes of plant natural products derived from chalcones are also discussed. Evolutionary mechanisms of type III PKS divergence are considered, as are the biological functions and activities of each of the known and functionally divergent type III PKS enzymc families (currently twelve in plants and three in bacteria). A major focus of this review is the integration of information from genetic and biochemical studies with the unique insights gained from protein X-ray crystallography and homology modeling. This structural approach has generated a number of new predictions regarding both the importance and mechanistic role of various amino acid substitutions observed among functionally diverse type III PKS enzymes. PMID- 12636086 TI - Molecular analysis of cis-prenyl chain elongating enzymes. AB - Recent isolation of the gene for an undecaprenyl diphosphate synthase has disclosed the structures of many kinds of cis-prenyl chain elongating enzymes. Not only the primary structure but also the crystal structure of the cis prenyltransferase is totally different from those of trans-prenyl chain elongating enzymes. This review covers up to February 2002 and contains 72 references. PMID- 12636087 TI - The biosynthesis of shikimate metabolites. AB - This review covers the literature published during 2000 on the biosynthesis of compounds derived wholly or partly from intermediates on the shikimate pathway. Recent developments in the enzymology and genetics of the shikimate pathway arc also described. Enzymes involved in the biogenetic pathway to the aromatic amino acids are covered initially followed by sections dedicated to metabolites derived in some part from intermediates on the pathway. These include pyrrolnitrin. violacein. indole-3-acetic acid, coumarins, lignans, lignin, tannins, melanin, flavanoids, ubiquinone, TOPA quinone, PQQ, and tropanes. PMID- 12636088 TI - The C7N aminocyclitol family of natural products. AB - This review covers microbial secondary metabolites classified in the family of C7N aminocyclitols, a relatively new class of natural products that is increasingly gaining recognition due to their significant biomedical and agricultural uses. Their discovery and structure determinations, their biosynthetic origin, biological properties, chemical synthesis, as well as their further development for pharmaceutical uses are described. The literature from 1970 to July 2002 is reviewed, with 269 references cited. PMID- 12636089 TI - Endocrine effects in the hazard assessment of drugs used in animal production. AB - Criteria of toxicological assessment are currently reviewed to ensure an adequate protection to susceptible groups, such as infants and children; in particular, concern arises about altered endocrine homeostasis in the developing organism, eliciting possible persistent effects such as reproductive disturbances and increased risk of tumours in target organs. Such items are obviously relevant also for veterinary drugs, whose main safety issue is potential lifetime exposure to residues. Two groups of chemicals, nitroimidazoles and imidazole antifungals, are reviewed as examples of the relevance of endocrine toxicity to the hazard assessment of compounds used in animal production. Nitroimidazoles are metabolized into genotoxic intermediates; in rodents they induce testicular toxicity and carcinogenicity. In particular, mammary neoplasms, mostly fibroadenomas, are consistently induced in rats as the most important effect in chronic studies; this may hint to possible endocrine-related mechanisms. Accordingly, evidences on other chemicals (e.g., triazines) show mammary tumorigenicity in rodents associated to hormonal alterations. In fact, nitroimidazoles affect synthesis of both pituitary and steroid hormones in vitro and rise progesterone and FSH levels in rats; also, limited clinical data in humans indicate endocrine-related effects. Overall, nitroimidazoles appear to affect the endocrine balance; however, the actual importance of such alterations, especially in regards to rat mammary tumours, has yet to be clarified. Imidazole antimycotics are broad-spectrum inhibitors of steroid synthesis; accordingly, diverse reproductive and developmental alterations are observed, depending on age and sex of animals exposed. Effects include pregnancy loss, delayed pup growth as well as reduced weight of androgen-dependent tissue; however, it is still difficult to identify the most susceptible biological phase. Overall the potential for inducing endocrine-related alterations should be carefully evaluated also for drugs used in animal production. A screening battery should produce a distinct fingerprint for each major endocrine activity, thus targeting longer-term tests (such as the two-generation study) on the most relevant endpoints. Moreover, the validation of molecular approaches would contribute to a biologically-based evaluation, by providing insights on such items as early effects and species-specificity. PMID- 12636090 TI - Primary hepatic endocrinomas (carcinoids and variant neoplasms). A statistical evaluation of 126 reported cases. AB - This study was undertaken to evaluate the statistical results of a large collected series of primary hepatic endocrinomas including carcinoids from extensive clinicopathologic aspect, and to provide the information to investigators engaged in this particular field of research. Primary hepatic endocrinomas variously termed as endocrine cell carcinoma (ECC), neuroendocrine carcinoma (NEC), carcinoid, and other carcinomas with endocrine components (CECs), have been rarely reported in literature. A total of 126 reported cases were registered in the Niigata Registry for gut-pancreatic endocrinomas. They consisted of a carcinoid group with typical carcinoids (n=82) and atypical carcinoids (n=19) and another group with other variant endocrinomas, briefly expressed as the variant group (n=25) in this study. The number of females was larger than that of males in the carcinoid group (M/F=0.58) and reversely males were predominant in the variant group (M/F=1.78). The average age was considerably older in the carcinoid group as compared to that in the variant group (53.9 years vs 48.3 years). The carcinoid group (either typical, atypical or both) showed a significant lower value than the variant group in metastasis rates in the large (over 50 mm) tumor series (43.3% vs 75.0%: P<0.05), and higher than the latter in serotonin positivity demonstrated by immunohistochemistry (37.0% vs 11.1%). While no significant difference was present between the two groups, cases with large tumors over 50 mm were found at a high rate (P<0.01) in approximately three fourths of each group, i.e. 76 out of a total of 100 cases recorded, and the liver was more often involved by metastasis than lymph nodes in the carcinoid group (32.7% vs 19.8%: P<0.05) though no significant difference was noted between these two sites in the variant group. The five-year survival rates calculated by the Kaplan-Meier method showed better results in the carcinoid group than in the variant group but with no statistically significant difference (92.4% vs 68.1%). A close similarity found in many aspects of the two groups suggested endocrinomas in the present study apparently belong to one family of CECs or briefly "carcinoid family". Regarding the more detailed statistical analyses, further accumulation of well-documented cases is needed. PMID- 12636091 TI - Sentinel node in gastric cancer surgery. AB - Sentinel Node (SN) biopsy studies have been recently applied to gastric cancer. In this series, 40 selected patients operated for gastric adenocarcinoma located in the lesser curvature and/or anterior wall of the body and antrum, underwent an intraoperative dye lymphography. The lymphatic ducts and nodes were visualized and a SN was evidenced in all cases. This was removed and a frozen section examined. In all cases a radical D2-3 gastrectomy was performed and histology, molecular biology, RT-PCR research of micrometastases (CEA-mRNA), were determined on the specimens. Correlations between T and histological status of SN and regional nodes were done. In 16 cases the SN was negative and all the resected regional nodes were negative too. In 15 cases the SN node was positive and other nodes in other stations were found to be positive as well. In 2 cases the SN was negative but other nodes, in the same stations and in others, were positive (false negative = 5%). In 7 cases the SN was the only node in which metastases occurred, 3 demonstrated by conventional histology and 4 detected by RT-PCR. In these 7 cases the SN was the only involved node out of all resected nodes, thus demonstrating to be the real first node along the lymphatic routes from the tumour. This experience seems to confirm the existence of a Sentinel Node and that each gastric adenocarcinoma has its own lymphatic basin in which metastasis can occur. Although a prudent attitude towards the indications resulting from these observations is required, in selected cases a controlled and tailored lymphoadenectomy could be adopted. PMID- 12636092 TI - Endogenous granulocyte colony-stimulating factor (G-CSF) levels in chemotherapy induced neutropenia and in neutropenia related with primary diseases. AB - Granulocyte-colony stimulating factor (G-CSF) and granulocyte macrophage colony stimulating factor (GM-CSF) are being administered to patients with neutropenia. However, little is known about the endogenous levels of both factors in these patients. We measured the endogenous G-CSF levels in patients with chemotherapy induced neutropenia (n=15, study group A), in patients who had not received chemotherapy with neutropenia caused by a number of primary diseases (n=14, study group B) and in healthy volunteers (n=15, control group). Both the study groups and the control group did not show any clinical or laboratory findings of infection. The G-CSF levels were elevated in patients following chemotherapy and in patients who had neutropenia without chemotherapy, but the mean G-CSF levels in patients with chemotherapy-induced neutropenia were significantly higher than in patients with primary diseases. The levels of endogenous G-CSF were also higher in both neutropenic groups, compared to the control group. In conclusion, endogenous G-CSF levels in chemotherapy-induced neutropenia were significantly higher than non-chemotherapy related neutropenia and controls. This may be explained as G-CSF synthesizing bone marrow stromal cells may be more affected in primary disease related neutropenia than in chemotherapy induced neutropenia. PMID- 12636093 TI - Fludarabine, ARA-C, idarubicin and G-CSF (FLAG-Ida), high dose ARA-C and early stem cell transplant. A feasable and effective therapeutic strategy for de novo AML patients. AB - Forty-three consecutive patients with de novo and untreated non M3 AML aged 60 or less entered the study. The mean age of patients was 50 (range 15-60). The induction regimen (FLAG-Ida) included fludarabine (30 mg/sqm), Ara-C (2 g/sqm) on days 1-5, and idarubicin (10 mg/sqm) on days 1, 3, 5. G-CSF (300 mcg/day) was administered s.c. 12 hours before starting fludarabine and was continued for five days. HDT with stem cell rescue was planned for all patients in first CR after one course of high dose Ara-C (HDAC) consolidation and in good clinical conditions. Forty-two (98%) patients were evaluable for response. One patient died during induction (2%). CR was achieved in 35 patients (82%). Twenty-three patients, 66% of those achieving CR, underwent autologous (N = 17) or allogeneic (N = 6) transplantation. With a median follow up of 24 months, the average median duration of CR is 17 months (range 3-66) and the median survival is 20 months (range 1-83). Overall the 5 year projected disease free survival (DFS) and overall survival (OS) were 37% and 43%, respectively. Among patients who underwent stem cell transplantation DFS and OS were 53% and 69%, respectively. The median time to PMN recovery (> 0.5 x 10(9)/l) was 17 days (range 10-28) and 50 x 10(9)/l platelets were reached at a median of 17 days (12-38). In conclusion FLAG-Ida regimen is effective, low toxic and improves feasibility of stem cell transplant. PMID- 12636094 TI - TNF-alpha levels in patients with malignant tumors after hyperthermic isolated regional perfusion. AB - Hyperthermic isolated regional perfusion is an alternative method for the treatment of malignancies especially sited in the pelvic region and extremities. The perfusion is performed via the extracorporeal system with a pump flow and chemotherapeutic agents and some cytokines, like tumor necrosis factor, may be added. It's well known that in the ischemia-reperfusion injury, hemorrhagic necrosis and cell death occurs and these cytokines are produced endogenously. In this study the endogenous TNF-alpha levels before, during and after hyperthermic isolated regional perfusion were compared in 16 patients with malignant tumors. The TNF-alpha levels were determined from the blood samples taken systemically before and after perfusion, and from perfusate at the 15th, 30th, 45th minute. The 15th minute samples were the ones where vascular clamps were applied to the vessels before starting the perfusion. TNF-alpha levels between the 15th minute samples and the blood samples that were taken systemically before and after perfusion were found statistically significant. The cause of this was suggested to be the ischemia-reperfusion injury in this period. It was shown in this study that high endogenous TNF-alpha levels and good clinical results could be achieved with hyperthermic isolated regional perfusion in the treatment of malignant tumors. PMID- 12636095 TI - Quantitative morphometrical investigation of basal cell layer in laryngeal premalignant lesions. AB - Histological diagnosis of laryngeal dysplasia is quite subjective. Since morphometry is highly reproducible, this method was applied to compare shape and size variations of the basal nuclei of the laryngeal epithelium in normal, laryngeal intraepithelium neoplasia (LIN) and invasive carcinoma to assess the reliability of light microscopic criteria used in grading dysplasia according to Friedman classification. Morphometrical analysis was carried out by Shape Analytical Morphometry (S.A.M) system. The logical architecture assumes that each irregular shape contains elements of two distinct logical domains: gross distortions that interest the contour and its local perturbations. These features were investigated separately by analytical procedures to acquire independent parameters both on the logical level and the numerical one. The nuclear area significantly increased from normal to carcinoma (p<.001). The increasing of the nuclear area was evident also in LIN I. Nuclear distortions were present in LIN II and LIN III. The highest nuclear contour irregularities were found in LIN III. Multivariate analysis showed a difficulty in discriminating various grades of dysplasia, especially between LIN I and LIN II (31% of error). In conclusion, our results indicate that nuclear pleomorphism of the basal cells layer, using a unique evaluator, is an unsatisfying criterion to distinguish moderate dysplasia. PMID- 12636096 TI - Palliative treatment of esophageal tumors. AB - In malignant dysphagia expandable metal stents are commonly used as palliative treatment, but early and late complications and the improvement of dysphagia have not been well described. This report summarizes our experience with expandable metal stents for malignant dysphagia. From 1995 to 2000, we placed 38 metal stents in 36 patients with malignant dysphagia from unresectable esophageal cancer (94.4%). Dysphagia scores, complications and modality of reintervention were evaluated. Dysphagia scores decreased from 3.2 before the stent placement to 2. Immediate complications occured in one patient because of severe pain, it was not possible to perform endoscopic treatments. Other complications included tracheoesophageal fistula (2 patients), tumor overgrowth (5 patients), new stent placements (2 patients), dislocation (2 patients). In conclusions expandable metal stents are safe and effective in the treatment of malignant dysphagia. PMID- 12636097 TI - Alleviation of cyclophosphamide-induced urotoxicity by naturally occurring sulphur compounds. AB - Treatment of Swiss albino mice with naturally occurring sulphur compounds such as Diallyl sulphide (DAS) (500 microg/dose/animal, i.p) and Diallyl Disulphide (DADS) (250 microg/dose/animal, i.p) for five days along with cyclophosphamide (CTX) (1.5 mmol/kg.body.wt; i.p) reduced the CTX induced urotoxicity. Morphological analysis of the urinary bladders of the CTX treated group showed severe inflammation and dark colouration whereas the sulphur compounds treated group showed almost normal bladder morphology. Treatment with DAS and DADS was found to reduce the urine urea N2 level DAS (15.31+/-0.01 g/l), DADS (12.38+/ 0.17 g/l) 4 hrs after CTX administration which was elevated in the CTX alone treated group (26.87+/-1.86 g/l). Urine protein level, which was enhanced drastically (8.66+/-0.47 g/l) after the CTX treatment, was significantly reduced (2.9+/-0.25 g/l) by the treatment with DADS. Similarly GSH content (which was drastically reduced by the CTX administration) in both bladder (0.87+/-0.1 nmol/mg protein) and liver (2.47+/-0.6 nmol/mg protein) was enhanced significantly (P<0.001) by the treatment with DAS and DADS both in bladder (DAS- 1.56+/-0.17 nmol/mg protein; DADS--2.65+/-0.21 nmol/mg protein) and liver (DAS- 5.30+/-0.07 nmol/mg protein; DADS--6.93+/-0.06 nmol/mg protein). Histopathological analysis of the bladder of the CTX alone treated group showed severe necrosis of the tissue whereas the sulphur compounds treated group showed normal bladder pathology. PMID- 12636098 TI - Antiproliferative activity of some cis-/trans-platinum(II) complexes on HeLa cells. AB - Purpose of this work was to synthesize several cis-/trans- isomer pairs of the platinum(II) complexes, and study the extent and the mode of their antiproliferative activity on HeLa cells. Six platinum(II) isomer pairs have a general formula cis-/trans-[PtA2X2], where A is ligand: ammonia (NH3), pyridine (Py); and X is ligand: chloride ion (Cl-), bromide ion (Br-), iodide ion (I-), thiocyanato ion (SCN-); four compounds have different structural formulas, and these are cis-/trans-[Pt(NH2OH)2(NH3)2]Cl2, and cis-/trans-Pt(Gly)2, where Gly is bidentate glycinato ligand. Results of the MTT assay, showed that six cis- and one trans-platinum(II) complexes exhibited cytotoxicity (IC50) ranging between 5 and 33 microM. Most of the cis-platinum(II) isomers caused significant alteration of cell cycle phases progression, and induced apoptosis in degree that varied among different compounds, as evaluated using flowcytometry and morphological study. Spectrophotometric analysis (AAS) indicated that there is no correlation between intracellular platinum(II) accumulation and cytotoxicity of tested complexes. PMID- 12636100 TI - Correlation between peri-operative serum lactate levels and outcome in pancreatic resection for pancreatic cancer, preliminary report. AB - This study was undertaken in order to verify if peri-operative serum lactate level changes, resulting from manipulation of the splanchnic circulation during pancreatectomy, reflected clinical outcome in twenty patients (9 males-11 females age 33 to 76) operated for pancreatic cancer. Lactate levels were evaluated at the beginning of the procedure (T0), after surgical manipulation before pancreatic resection (T1), after resection (T2), and 24 hours post-op. (T3). Furthermore, to highlight possible hemodynamic instability that could contribute to altered lactate clearance, mean arterial pressure (MAP) and central venous pressure (CVP) were continuously monitored during the study period. Peri operative mortality within 60 days after surgery, Intensive Care Unit (ICU) length of stay, and peri-operative complications were the main indicators investigated in order to evaluate the impact of serial lactate levels in this patient population. Hyperlactatemia observed peri-operatively during pancreatic resection for cancer is significantly correlated with peri-operative mortality and also with longer ICU length of stay. Though, due to the relatively small number of the patients, more extensive investigation is needed in order to confirm such interesting preliminary data. PMID- 12636099 TI - Differential diagnosis of atypical epithelium of biopsied gastric mucosa using immunostaining of Ki-67, p53, hMLH1 and MDM2 expression. AB - Gastric biopsy specimens were classified from category 1 (C1) to C5 according to the Vienna classification of gastrointestinal epithelial neoplasia. Fifty cases of C1, 40 cases of C2, 29 cases of C3, 19 cases of C4-1, 20 cases of C4-2+3, and 24 cases of C5 were selected and the expression of Ki-67, p53, hMLH1, and MDM2 was examined immunohistochemically to obtain useful data for the differential diagnosis of controversial cases of C2 and C4. The C2 cases, which did not show definitive neoplasia, were divided into two groups: one group had regenerative epithelium showing a relatively high degree of atypia (regenerative atypia: RA) and the other had carcinoma showing a mild degree of atypia (carcinoma of low grade atypia: CLGA). There were 0/23 RA and 12/17 CLGA cases that were positive for p53. Ki-67-positive cells were localized in the mitotic cell zone in RA, whereas they were irregularly distributed in CLGA. None of the cases of high grade adenoma (C4-1) showed p53 expression, whereas 19 out of 44 well differentiated adenocarcinoma cases (C4-2+3 and C5) were positive for p53. Ki-67 positive cells were localized in the mitotic cell zone in C4-1, whereas they were irregularly distributed in C4-2+3. hMLH1 was expressed in the proliferative cell zone of the normal foveolae, in all of the cases of adenoma (C3, C4-1) and in most cases of adenocarcinoma (C4-2,3 and C5). However, in 3 cases of C4-2+3, which showed a relatively mild degree of cellular atypia and were difficult to differentiate from high-grade adenoma (C4-1), the neoplastic cells were completely negative for hMLH1. MDM2 showed no characteristic expression in any gastric lesion. In conclusion, p53-positive cases of C2 and C4 were highly suggestive of carcinoma. Even if the case was negative for p53, the irregular distribution pattern of Ki-67-positive cells was highly suggestive of carcinoma. hMLH1-negativity in the C4 cases is strongly suggestive of carcinoma and this may be an useful marker for differentiating C4-2+3 from C4-1. PMID- 12636101 TI - Carcinoembryonic antigen mRNA in abdominal cavity as a useful predictor of peritoneal recurrence of gastric cancer with serosal exposure. AB - Peritoneal dissemination is the most frequent type of recurrence in patients with gastric cancer with serosal exposure, irrespective of whether they have undergone curative gastrectomy. The purpose of this study was to establish a method to detect micrometastatic cells in the abdominal cavity and predict peritoneal recurrence in patients with such gastric carcinomas. A total of 86 patients with gastric carcinoma, undergoing gastrectomy, were examined. Reverse transcriptase polymerase chain reaction (RT-PCR) assay was used to detect carcinoembryonic antigen (CEA) mRNA in abdominal lavage fluid. Twenty-four cases without serosal exposure were negative, while all 13 cases with macroscopic peritoneal dissemination were positive for CEA mRNA. Among the 49 cases with macroscopic serosal invasion and without peritoneal metastasis, cancer cells were detected in 27 cases with RT-PCR while in only 6 cases with conventional cytology. All cytologically-positive cases were also positive for CEA mRNA. Among the 27 CEA positive cases, 15 patients (56%) relapsed with peritoneal metastasis within 12 months after gastrectomy. In contrast, none of the 22 CEA-negative cases had peritoneal recurrence within 16-60 months of observation, whereas in 43 cytologically-negative cases, 10 patients relapsed with peritoneal recurrence. As compared with conventional cytological examination, this method would be clinically more beneficial for detecting free cancer cells in the peritoneal cavity and for predicting peritoneal recurrence in gastric carcinoma with serosal invasion. PMID- 12636102 TI - Detection of peritoneal micrometastases by reverse transcriptase-polymerase chain reaction targeting carcinoembryonic antigen and cytokeratin 20 in colon cancer patients. AB - Peritoneal recurrence after curative resection of malignant tumor with negative cytology is considered to be caused by microscopic dissemination of the exfoliated cancer cells from primary tumors to serosal surfaces at the time of operation, not detectable with conventional diagnostic tools. We applied the reverse transcriptase-polymerase chain reaction (RT-PCR) for carcinoembryonic antigen (CEA) and cytokeratin 20 (CK 20) to detect micrometastatic foci in the peritoneal cavity of colon cancer patients. Cytological samples taken by peritoneal lavage from a series of 79 colon cancer patients were analyzed microscopically, for CEA levels, and by RT-PCR analysis using nested primers for CEA and CK 20. Cases with both CEA and CK 20 signals were defined as PCR positive. This RT-PCR method proved both sensitive (1 tumor cell/10(6) non-tumor cells on preparation of serial colorectal cancer cell dilutions) and specific (no false positive results, 0/23 tested in our control experiment). Intraperitoneal micrometastatic cells were detected in peritoneal lavage 7.6% by cytology, 17.7% by CEA levels, and 24.1% by RT-PCR (significantly higher than by cytology: p=0.0046). RT-PCR detection rate increased in parallel with pathological depth of tumor invasion, and also a pathological stage-dependence was suggested according to the tumor-node-metastasis classification of the International Union Against Cancer. Our results suggest that CEA and CK 20 mRNA identification by RT-PCR appeared to be reliable and may be useful for early diagnosis in peritoneal dissemination of colon cancer. PMID- 12636103 TI - Studies with black tea and its constituents on leukemic cells and cell lines. AB - The anticancer effect of black tea (BT) and its polyphenols theaflavin (TF) and thearubigin (TR) has been evaluated on U-937 cell line, a myeloid leukemic cell line and on leukemic cells isolated from peripheral blood of chronic myeloid leukemia (CML) patients. In both types of cells, cell growth inhibition was observed 24 hrs after treatment with BT, TF and TR. MTT assay showed growth inhibition of metabolically active cells and inhibition of DNA synthesis was observed by 3H-Thymidine incorporation after treatment with the compounds. In all cases TF and TR were more effective than BT, suggesting that these are possibly the active components in BT responsible for its antileukemic activity. Superoxide dismutase (SOD), a free radical scavenger, was found to be increased by TF, whereas BT and TR lowered the level in comparison to the control. The present study is the first report of antileukemic effect of BT and its polyphenols. PMID- 12636105 TI - Effects of bile acids on 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine-induced aberrant crypt foci and DNA adduct formation in the rat colon. AB - The effects of deoxycholic acid (DCA) and ursodeoxycholic acid (UDCA) on 2-amino 1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-induced aberrant crypt foci (ACF) in the rat colon were examined. The effect of these bile acids on DNA adduct formation by PhIP in the colon was then analyzed, since the main action of PhIP is the formation of DNA adducts and subsequent gene mutations. For the ACF study, male F344 rats were administered PhIP-HCl (75 mg/kg, 10 doses) by gavage, and a diet containing bile acid (0.4% DCA or UDCA) was provided from 3 days before the first dose of PhIP for 8 weeks. The mean number of ACF per colon of DCA, UDCA and controls were 9.9, 2.4 and 5.5, respectively. The ACF number was significantly increased by DCA and decreased by UDCA (P<0.001). To examine the effect of bile acids on DNA adduct formation, male F344 rats were fed a diet supplemented with bile acids (0.1 or 0.4% of DCA and UDCA) 7 days prior to the PhIP administration. All rats were administered a single dose of PhIP-HCl (50 mg/kg) by gavage and sacrificed 48 hours later. DNA adduct levels of the 0.1% UDCA, 0.1% DCA and controls were 2.93 (adducts/10(7) nucleotides), 2.65 and 1.10, respectively. Those of 0.4% UDCA, 0.4% DCA and controls were 1.64, 1.30 and 1.00, respectively. The PhIP-DNA adduct level was significantly increased by administration of 0.1% UDCA, 0.1% DCA (P<0.05) and 0.4% UDCA (P<0.01). The increasing effect of both DCA and UDCA on PhIP-induced DNA adduct formation was unexpected, and was not directly associated with ACF formation. PMID- 12636104 TI - An immunohistochemical study of cyclooxygenase (COX)-2 expression in endocrine tumors of the pancreas. AB - Cyclooxygenase (COX) is a key rate-limiting enzyme in prostaglandin biosynthesis. There are two isoforms of COX, referred to as COX-1 and COX-2. COX-2, an inducible form of COX, is found to be overexpressed in various neoplasms and is believed to play an important role in tumorigenesis and tumor development. In this study, we investigated expression of the COX-2 protein in human endocrine tumors of the pancreas (N=23; 6 insulinomas, one glucagnoma, 2 gastrinomas, and 14 non-functioning tumors) using immunohistochemistry. Strong COX-2 expression was confirmed in normal islet tissue as previously reported. COX-2 immunoreactivity was detected in 65% (15 out of 23) of these tumors with a moderate to strong intensity. In all nine functioning tumors, COX-2 expressions were preserved with the weak or strong intensity. In contrast, COX-2 was present in 6 out of 14 nonfunctioning tumors. The correlation between COX-2 expression and their function was significant (p<0.05). We found that expression of this enzyme was detected in 11 out of 15 benign tumors and in 4 out of 8 malignant tumors, respectively. Our results suggest that COX-2 may play an important role in the endocrine function of islet tumors. Additionally, malignancy was not related to COX-2 expression. PMID- 12636106 TI - Effect of Withania somnifera on cell mediated immune responses in mice. AB - Effect of Withania somnifera on the cellular immune responses (CMI) was studied in normal as well as tumour bearing animals. Administration of Withania extract was found to enhance the proliferation of lymphocytes, bone marrow cells and thymocytes in responses to mitogens. Both PHA and Con A mitogens along with Withania treated splenocytes, bone marrow cells and thymocytes could stimulate proliferation twice greater than the normal. Withania treated splenocytes along with the mitogen LPS (10 microg/ml) could stimulate the lymphocyte proliferation six times more than the normal. Natural killer cell activity (NK) was found to be enhanced significantly in both the normal and the tumour bearing group and it was found to be earlier than the control (48.92% cell lysis). Antibody dependent cellular cytotoxicity (ADCC) was found to be enhanced in the Withania treated group on the 9th day (65% cell lysis). An early Antibody dependent complement mediated cytotoxicity (ACC) was observed in the Withania treated group on day 13 (47% cell lysis). PMID- 12636107 TI - In situ RT-PCR detection of hepatitis C virus genotypes in Chinese patients with hepatocellular carcinoma. AB - The correlation of hepatitis C virus genotypes and the development of hepatocellular carcinoma is still controversial. The most pertinent studies are conducted by seroepidemiological methods. It has been suggested that tissue RT PCR may be more efficient than serological testing for hepatitis C virus RNA. The purpose of this study was to investigate the infectious status of hepatitis C virus genotypes in hepatocellular carcinoma tissue for revealing the role of hepatitis C virus genotypes in carcinogenesis of hepatocellular carcinoma. Hepatitis C virus genotypes in cancerous and noncancerous liver tissues of 95 Chinese patients with hepatocellular carcinoma were analyzed by a modified type specific in situ reverse transcription polymerase chain reaction (RT-PCR) method. Hepatitis C virus genotypes were simultaneously examined by Okamoto's method using extracts from hepatocellular carcinoma tissues. The hepatitis C virus genotypes in hepatocellular carcinoma tissues were found including 1b, 2a, mixed type (1b+2a) and 3a in 13, 23, 2 and 4 samples respectively. There was no significant difference in either the positivities between 1b (13.7%, 13/95) and 2a (24.2%, 23/95) (P>0.05) or the staining intensity and distribution of positive cells between 1b and 2a (p>0.05). No statistically significant difference was found between the two virus genotypes in main clinico-pathologic parameters of hepatocellular carcinoma. The localizations of hepatitis C virus RNA-positive signals were mainly cytoplasmic (22/36) in noncancerous regions, but nucleocytoplasmic (19/36) or nuclear (3/36) in cancerous regions. The positive nucleocytoplasmic/nuclear signals of genotype 1b (11/13) in hepatocellular carcinoma were seen more frequently than those of genotype 2a (11/23) (P<0.05). The discordant hepatitis C virus 2a and 1b positivities in serum samples and in cancerous tissues of hepatocellular carcinoma patients were also found, suggesting that the detection of serum hepatitis C virus genotypes in the patients with hepatocellular carcinoma may not reflect the actual status of hepatitis C virus genotypes in cancerous tissue. The pathogenetic significance of existence of hepatitis C virus gene in the nucleus of hepatocytes and malignant cells needs further investigation. PMID- 12636108 TI - The SEIRA spectroscopy data of nucleic acids and phospholipids from sensitive- and drug-resistant rat tumours. AB - It is known that tumour progression towards drug resistance is one of the main factors resulting in the failure of cancer treatment. As tumour progression is based on the genetic instability, the study of the structure of nucleic acid from tumour cells is of great importance both for basic knowledge and for biomedical application. We applied surface enhanced infrared absorption spectroscopy (SEIRA) of nucleic acids on gold substrate and essentially increased the sensitivity of IR spectroscopy. We observed numerous changes in infrared spectra of DNA from sensitive and resistant cells that reflect drastic changes in molecular structure of DNA from tumour cells. The DNA from resistant cancer cells could be characterised as rigid structure, the structure of DNA from sensitive cancer strain seems to be flexible and after application of anticancer drugs drastically changes and approaches to the structure of helix forms. The molecular structure of lipids from resistant and sensitive cancers after application of anti-tumour drugs is also modified. Thus, we observed a disordering in the lipid chain packing from resistant cells after application of cisplatin and, in some cases, formation of phospholipid-Pt complex. PMID- 12636109 TI - Complete response of Sister Mary Joseph Nodule from gastric adenocarcinoma treated with combination chemotherapy of low-dose S-1 and cisplatin. AB - A case of an unresected, advanced gastric cancer with Sister Mary Joseph nodule was presented. It was treated with new combination chemotherapy of low-dose S-1 and cisplatin producing complete response of periumbilical metastasis. Few treatments are efficacious for umbilical invasion of peritoneal dissemination. A complete response for Sister Mary Joseph nodule from gastric adenocarcinoma has not been ever reported. PMID- 12636110 TI - Adenocarcinoma on horseshoe kidney studied with computed tomography. AB - Horseshoe kidney is a congenital anomaly of the upper urinary system frequently associated with atypical vascularization, mostly asymptomatic, usually undetected until the onset of infectious, obstructive or neoplastic complications. Although intravenous pyelography and US are useful tools, the role of CT is of vital importance especially in the pre-operative planning of related complications and in those cases where other techniques have correctly diagnosed the disease but have missed the visualization of an underlying malformation. We report here two cases of adenocarcinoma in two patients with a horseshoe kidney, where CT has clearly depicted both the malformation and the associated pathologic findings. PMID- 12636111 TI - Gastrointestinal stromal tumor (GIST) and ulcerative colitis. AB - A 57 year old woman, affected by Ulcerative Colitis (UC) in remission, was admitted to our unit with a history of episodic melena and progressive anemia. Computed tomography (CT) of the abdomen revealed a solid mass in the upper left pelvic cavity. After surgical laparotomy, the mass showed histological characteristics of a gastro intestinal stromal tumor. This report describes a GIST in a patient with extensive UC, which was in remission at the time of diagnosis. To our knowledge, this is the first report of an association of this type. PMID- 12636112 TI - Histopathological diagnosis of biopsy samples from early esophageal carcinoma. AB - Carcinoma of the esophagus is frequently diagnosed in advanced clinical stages. When an esophagic carcinoma has infiltrated the submucosa or the muscular or serosa, metastases are a common finding. Thus, early diagnosis and opportune treatment are vital for patients with this type of neoplasm. Timely diagnosis can be done through endoscopic or X-ray studies and confirmed through a histopathological study by directed biopsy. We presently report the case of a 65 year old man with precedents of achalasia who underwent an endoscopic study using the Lugol staining technique for suspected malignant lesion classified as 0-IIc. After two biopsies it was diagnosed as early carcinoma of the esophagus and was subjected to mucosectomy. Histopathological findings are reviewed at architectural and cellular level and are essential to establish the diagnosis of early neoplastic lesions of the esophagus epithelium. These cellular changes are corroborated by immunohistochemical studies with nuclear expression of p53. The relevant literature was reviewed and experiences by Japanese and North American pathologists compared with emphasis on the need for multidisciplinary management to make an early diagnosis by endoscopic studies, Lugol staining, X-rays, biopsy and conservative treatment based on mucosectomy. PMID- 12636113 TI - The knowledge gap: difficult aspects of communication in oncology. PMID- 12636114 TI - Mens et manus: striking a balance between skill and reason. PMID- 12636115 TI - Gold foil restorations. PMID- 12636116 TI - More gold foil. PMID- 12636117 TI - Pulpal death. PMID- 12636118 TI - Do you prefer print or online publications? PMID- 12636119 TI - Spyware: it's all about you. PMID- 12636120 TI - Light augments tooth whitening with peroxide. AB - BACKGROUND: The authors tested the adjunctive use of light with a 15 percent peroxide gel as a single-visit, in-office tooth whitening system. METHODS: Subject (N = 87) with stained (> shade D4, Vita Zahnfabrik, Bad Sackingen, Germany) anterior teeth were randomly assigned to test (peroxide and light), peroxide control (peroxide gel) or light control (placebo gel and light) groups and were treated for one hour. The researchers evaluated tooth shade, color and subject response at baseline and posttreatment and at three and six months posttreatment. RESULTS: The initial shade unit reduction of combined light and peroxide treatment (8.4) was greatest compared with that of peroxide alone (5.9) and of light alone (4.9). Approximately 88 percent of these effects persisted for six months. Lightness was increased and yellowness decreased to a significantly greater extent in the test group than in either control. These findings were corroborated by subject evaluation. One week after treatment, moderate to greatly increased tooth sensitivity occurred in 20 percent of test subjects, 21.7 percent of peroxide control subjects and none of the light control subjects. Neither tooth sensitivity nor gingival redness was present at the three- and six-month visits. CONCLUSIONS: Peroxide and light treatment significantly lightened the color of teeth to a greater extent than did peroxide or light alone, with a low and transient incidence of tooth sensitivity. CLINICAL IMPLICATIONS: Light can increase the tooth-whitening effect of peroxide, thereby increasing the effectiveness of tooth-whitening procedures. PMID- 12636121 TI - Three restorative materials and topical fluoride gel used in xerostomic patients: a clinical comparison. AB - BACKGROUND: The authors compare the incidence of recurrent caries around two glass ionomer restorative materials and one amalgam material. METHODS: The authors placed 111 restorations in nine xerostomic patients. Patients were given oral hygiene instructions and fluoride gel to use daily. The authors evaluated the restorations clinically and with photographs and impressions at six months, one year and two years according to criteria for marginal adaptation, anatomical form, caries in adjacent tooth structure and caries at the cavosurface margin. The authors divided patients into users and nonusers of fluoride. RESULTS: At the two-year recall appointments, the authors evaluated 95 (86 percent) of the 111 restorations. They analyzed two-year data using an analysis of variance for repeated measures and Fisher's post hoc test. The study results showed no significant differences among materials in regard to caries at the cavosurface margin among fluoride users. However, among fluoride nonusers, patients with amalgam restorations had a significantly higher incidence of caries at the cavosurface margins than did patients with either of the two glass ionomer restorations. The authors found no statistically significant difference between restorations with regard to marginal integrity or anatomical form. However, one patient exhibited failure of glass ionomer restorations owing to material loss. CONCLUSIONS: Less caries developed at the margins of glass ionomer restorations compared with amalgam restorations in xerostomic patients who did not routinely use a neutral topical sodium fluoride gel. CLINICAL IMPLICATIONS: Fluoride releasing materials may reduce caries surrounding restorations in high-risk patients who do not routinely use topical fluoride. Patients who routinely used topical fluoride gel did not develop recurrent caries, and clinicians should encourage the use of fluoride gel on a daily basis. PMID- 12636122 TI - Allergic contact dermatitis in dental professionals: effective diagnosis and treatment. AB - BACKGROUND: Like other health care workers, dental professionals are at risk of developing allergic contact dermatitis, or ACD, after exposure to allergenic chemicals. Common allergens include antimicrobials, preservatives, rubber additives and methacrylates. CASE DESCRIPTION: The authors describe an orthodontic assistant with severe skin disease, whose symptoms included redness, cracking and bleeding that persisted for 10 years. The patient had previously received an incomplete diagnosis. After performing patch testing, assessing symptoms and evaluating the patient's medical history, the authors diagnosed ACD resulting from exposure to several dental allergens. The patient received appropriate treatment and counseling to better manage her allergies; this resulted in resolution of all symptoms and averted permanent occupational disability. CLINICAL IMPLICATIONS: Not all skin reactions are related to gloves or natural rubber latex. Dental professionals should be aware of common chemical allergens, symptoms of ACD and the appropriate treatment of occupational skin disease. PMID- 12636123 TI - Lingual nerve damage due to inferior alveolar nerve blocks: a possible explanation. AB - BACKGROUND: An explanation for the predominance of injuries to lingual nerves over those to inferior alveolar nerves as a result of inferior alveolar nerve blocks may be due to the nerves' fascicular pattern. A unifascicular nerve may be injured more easily than a multifascicular nerve. METHODS: The authors unilaterally dissected lingual and inferior alveolar nerves from 12 cadavers. They cut the specimens 2 millimeters above the lingula for both the lingual nerve and inferior alveolar nerve and opposite the site of the middle of the third molar for the lingual nerve, and they counted the number of fascicles at each site. RESULTS: For the lingual nerve at the lingula, the mean number of fascicles was three (range, one to eight). Four of the 12 nerves (33 percent) were unifascicular at this point. Opposite the third molar, the lingual nerve had a mean of 20 fascicles (range, six to 39). In every case, there were more fascicles in the third molar region than above the lingula in the same nerve. At the lingula, the inferior alveolar nerve had a mean of 7.2 fascicles (range, three to 14). CONCLUSION: This study may explain the observation that when an inferior alveolar nerve block causes permanent nerve impairment, the lingual nerve is affected about 70 percent of the time and the inferior alveolar nerve is affected only 30 percent of the time. In 33 percent of cases, the lingual nerve had only one fascicle at the lingula; a unifascicular nerve may be injured more easily than a multifascicular one. CLINICAL IMPLICATIONS: There is no known way to avoid the remote possibility of nerve damage resulting from an inferior alveolar nerve block. The lingual nerve may be predominantly affected because of its fascicular pattern. PMID- 12636124 TI - The diagnosis and management of recurrent aphthous stomatitis: a consensus approach. AB - BACKGROUND: Recurrent aphthous stomatitis, or RAS, is a common oral disorder of uncertain etiopathogenesis for which symptomatic therapy only is available. This article reviews the current data on the etiopathogenesis, diagnosis and management of RAS in a primary care setting. METHODS: The authors reviewed publications on Medline from 1995 through 2000, the period since the last major reviews were published. RESULTS: RAS may have an immunogenetic background owing to cross-reactivity with Streptococcus sanguis or heat shock protein. Predisposing factors seen in a minority include haematinic (iron, folate or vitamin B12) deficiency, stress, food allergies and HIV infection. While topical corticosteroids remain the mainstay for therapy, a number of other immunomodulatory modalities now are available. CONCLUSIONS: There is still no conclusive evidence relevant to the etiopathogenesis of RAS, and therefore therapy can attempt only to suppress symptoms rather than to address the basic issues of susceptibility and prevention. CLINICAL IMPLICATIONS: In the majority of patients, symptomatic relief of RAS can be achieved with topical corticosteroids alone, with other immunomodulatory topical agents or by combination therapy. PMID- 12636125 TI - Oral malodor. PMID- 12636127 TI - The etiology and prevalence of gingival recession. AB - BACKGROUND: Gingival recession in its localized or generalized form is an undesirable condition resulting in root exposure. The result often is not esthetic and may lead to sensitivity and root caries. Exposed root surfaces also are prone to abrasion. The purpose of this article is to describe the prevalence, etiology and factors associated with gingival recession. TYPES OF STUDIES REVIEWED: The authors reviewed cross-sectional epidemiologic studies of gingival recession and found that they correlated the prevalence of recession to trauma, sex, malpositioned teeth, inflammation and tobacco consumption. The recent surveys they reviewed revealed that 88 percent of people 65 years of age and older and 50 percent of people 18 to 64 years of age have one or more sites with recession. The presence and extent of gingival recession also increased with age. RESULTS: More than 50 percent of the population has one or more sites with gingival recession of 1 mm or more. The prevalence of gingival recession was found in patients with both good and poor oral hygiene. It has been proposed that recession is multifactorial, with one type being associated with anatomical factors and another type with physiological or pathological factors. Recession has been found more frequently on buccal surfaces than on other aspects of the teeth. CLINICAL IMPLICATIONS: Dentists should be knowledgeable about the etiology, prevalence and associating factors of gingival recession, as well as treatment options, so that appropriate treatment modalities can be offered to patients. Treatments for gingival recession include gingival grafting, guided tissue regeneration and orthodontic therapy. Such treatments typically result in esthetic improvement, elimination of sensitivity and a decreased risk of developing root caries. PMID- 12636126 TI - ADA members weigh in on perceptions of the ADA and association priorities. AB - BACKGROUND: The American Dental Association conducted the 2000 Membership Needs and Opinions Survey to evaluate the professional needs and personal perceptions of ADA members on selected topics. METHODS: A questionnaire was sent to a sample of 6,310 ADA members in January 2000 with follow-up mailings in February, March and April 2000. Data collection was completed in July 2000. The survey included questions on critical professional issues, perceptions of the American Dental Association and its priorities. A total of 3,558 surveys were received for an adjusted response rate of 59.5 percent. Only professionally active dentists who were ADA members were included in the sampling frame. RESULTS: Members evaluated statements about the American Dental Association, revealing their perceptions of the ADA on key issues. Findings showed strong support for the ADA Seal program, agreement that the ADA enhances the integrity and ethics of the profession and that the ADA is the premier professional association for dentists. Findings also provided information regarding the relative importance of ADA priorities by allocating "dues dollars" in $5 increments. The highest priorities were "providing continuing education to dentists," "lobbying members of Congress and federal regulatory agencies" and "influencing national public health policy." CONCLUSIONS: ADA members, both new and established dentists, had positive perceptions of the Association and its programs. Although there was general agreement between these groups concerning ADA priorities, there was a substantial difference between some subgroups (especially graduate students and federally employed dentists) and the overall membership on the issue of changes in the licensure process to facilitate dentists' freedom of movement. PRACTICE IMPLICATIONS: The Association should continue to take into account the membership's perceptions of the ADA and its priorities, as well as note issues of special interest to selected membership subgroups, in the planning and implementation of Association activities. PMID- 12636128 TI - Tap water filters. AB - Moen PureTouch filters remove impurities from tap water without removing fluoride. These carbon block filters consist of finely powdered activated carbon that is combined with a plastic binder material and heated to form a hollow cylinder. The blocks are further wrapped with material to improve performance and reduce clogging. The filters are available with different filtering capabilities (Table 1). The filters mount in the faucet spout or under the sink. PMID- 12636129 TI - Dental local anesthetics: alternative delivery methods. AB - BACKGROUND: The authors review four techniques for producing block and topical anesthesia that have been advocated as alternatives to conventional methods. The objective of this review is to examine the basis for each method's application in dentistry, including scientific studies where available. TYPES OF STUDIES REVIEWED: The authors reviewed both historical (1912 and 1923) and more recent (1977 to present) reports relating to specific local anesthetic delivery systems. While this article is not a comprehensive literature review, the clinical reports and double-blind scientific reports from peer-reviewed publications are specific to the various attributes of the delivery systems being described. In addition, the authors identify any claims lacking scientific validity. RESULTS: Several alternative local anesthetic delivery systems are clinically effective in producing local anesthesia. Potential adverse effects and complications do, however, differ from one technology to another, depending on the mechanical characteristics of the particular system. Finally, all of the alternative local anesthetic delivery systems represent additional costs when compared with those of conventional applications, and cost-effectiveness is an important factor to be considered when implementing these systems in a dental practice. CLINICAL IMPLICATIONS: Alternative local anesthetic delivery systems may offer some advantages over conventional techniques in certain situations, although the costs may outweigh the advantages in certain cases. PMID- 12636131 TI - For the dental patient... Treating periodontal disease: scaling and root planing. PMID- 12636130 TI - The best way to initiate a dental practice. PMID- 12636132 TI - Supporting the Federal Emergency Management Agency Rescuers: a variation of critical incident stress management. AB - The purpose of this article was to document the efforts of two military installations in facilitating Critical Incident Stress Management teams for Federal Emergency Management Agency Rescue and Recovery Units following the attacks on the World Trade Center on September 11, 2001. McGuire Air Force Base accepted the mission to support the rotating task forces with emotional support, and Fort Dix offered temporary lodging while the teams were in-bound and out bound to the "Ground Zero" site. A team, comprised of staff from both installations due to the configuration of professional and support staffs, does the Critical Incident Stress Management work. Both installations are within commuting distance of New York City and could provide adequate safety, security, and logistics to the teams. The classic crisis management models were not called into play, as that service was not asked for; however, it was clear to the McGuire Air Force Base/Fort Dix Critical Incident Stress Management team members that the rescue and recovery teams needed their physical presence. Many rescue organizations have their own "debriefing teams," yet it is believed that "outside teams" may be helpful as interim measures to demonstrate the universality of the process of coping with traumatic stress and grief management. It is hoped that this article honors the workers who came together to help in a very difficult time for our country. This article documents the interim measures that were taken for a group of people moving through crisis. PMID- 12636133 TI - Evaluation of the effect of flights in supersonic fighters on sinus rhythm variability parameters. AB - Heart rate variability is a widely used method to evaluate vagal and sympathetic influences on the heart. The purpose of this study was to assess the relationship of autonomic nervous system components' interaction in supersonic fighter pilots by means of a sinus rhythm variability analysis. The following frequency domain parameters were analyzed: low-frequency (LF) power spectral of 0.05 to 0.15 Hz (ms2); high-frequency (HF) power spectral of 0.15 to 0.45 Hz (ms2); LF and HF power spectral index; total spectral power (HF + LF); percent, LF distribution in total spectral power; percent HF, HF distribution in total spectral power; and an average heart rate per minute. The study comprised 30 healthy military pilots (age, 24-48 years) in whom heart rate variability parameters were determined 10 to 12 minutes before take off, 10 to 12 minutes after four successive flights, and 1, 2, and 12 to 16 hours after completing the flights. The control group was comprised of 20 healthy nonpilots (age, 24-49 years). The statistical significance was estimated by means of the following methods: Student's t test for normal distribution and a rank test for the non-normal one (between the groups of the same subjects); and the Cochran-Cox test for normal distribution and the Wilcoxon test for the non-normal one (between the groups of different subjects). Higher activity of sympathetic components of the autonomic nervous system was shown in pilots in the course of the first examination before flights as compared with the controls (higher LF, LF/HF, percent LF). Predominance of the sympathetic nervous system in pilots might reflect adaptation to repeated short term exposures to high values of accelerations acting in head-leg axis. PMID- 12636134 TI - The effect of driving distance from medical treatment facility and short-term management outcomes in Department of Defense beneficiaries newly diagnosed with human immunodeficiency virus. AB - Access to experienced providers is an important part of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome management, especially for patients who are just starting a new or complicated treatment regimen. To date, few studies have addressed the potential barrier of geographic distance in access to care in the HIV/acquired immunodeficiency syndrome population. We performed a retrospective study examining the effects of driving distance from a major Naval medical treatment facility on clinical outcomes for Department of Defense beneficiaries recently diagnosed with HIV. The outcomes studied include attainment of undetectable HIV-1 RNA levels and change in mean CD4+ T cell counts 6 months after initiation of management in 132 Department of Defense beneficiaries. We found no significant associations between distance from a major medical treatment facility and these clinical outcomes through 6 months of follow up. These findings suggest that stationing HIV-infected military personnel far from major Medical Treatment Facility does not interfere with initial successful HIV management. PMID- 12636135 TI - The incidence of eye injuries at three U.S. Army installations. AB - Studies addressing the impact of eye injuries on U.S. Army readiness and the effectiveness of countermeasures are lacking. In 1989, the U.S. Army Environmental Hygiene Agency undertook a prospective, multicenter study of military units in the peacetime Army at selected installations to define the incidence of eye injuries, assess associated variables, and identify targets for intervention. During the first 5 months of 1989 at Fort Bragg, North Carolina, Fort Campbell, Kentucky, and Fort Lewis, Washington, health care providers saw 103 new eye injuries with an overall rate of 14.18 injuries per 1,000 personnel annually. Almost one-half of the injuries (48 cases, 46.60%) occurred during on duty work and training. Most (93 cases, 90.29%) of the soldiers were not wearing eye protection when injured. This study identifies possible targets for intervention and provides a baseline against which the interventions of the last decade may be assessed for effectiveness. PMID- 12636136 TI - Depression in men with traumatic lower part amputation: a comparison to men with surgical lower part amputation. AB - This study aimed at comparing the prevalence of depression among traumatic and surgical amputees and examining the relationship between depression and sociodemographic/clinical characteristics of amputees. Participants were 49 patients with traumatic lower part amputation and 35 patients with surgical lower part amputation. The diagnosis of depression in each participating patient was confirmed by means of the Structured Clinical Interview for DSM-IV, Turkish version. The level of depression was assessed by using the Hamilton Depression Rating Scale. The prevalence of depression was 34.7% in the traumatic amputee group and 51.4% in the surgical amputee group (p > 0.05). In the traumatic group, depression was associated with time since amputation but not with other variables. However, in the surgical group, depression was associated with age, education level, marital status, economic status, time since amputation, and whether the patient was treated with prosthesis. Our data indicated that depression is a common clinical condition among amputees. Clinicians may be advised to schedule periodic contacts with amputees over long periods to identify those in need of psychiatric intervention. PMID- 12636137 TI - Medical support of the Sinai Multinational Force and Observers: an update, 2001. AB - The Multinational Force and Observers is an 11-nation coalition force with the mission of peacekeeping in the Sinai. It commenced operations in 1982 and continues today after two decades of successful enforcement of the 1979 Egyptian Israeli Treaty of Peace. The fielding of a medical support team for this mission was the first effort of its kind for the U.S. Army Medical Department, resulting in a uniquely organized medical unit. A U.S. Army physician heads the medical team, which includes nine other physicians from six different countries. The team provides health care for the 2,500-person coalition in the remote Sinai desert. This writing compares earlier medical support with that of August 2000 through July 2001 and describes medical conditions and problems encountered during the 12 month tour of duty. This article updates two previous articles on the subject published in Military Medicine in 1983 and 1991. PMID- 12636139 TI - Proposal of a model for medical records for international disaster relief operations. AB - The triage of a large number of victims in an international disaster relief operation is very important. To ensure the correct way of registration of patients and effective triage and treatment, useful medical records are necessary. The authors proposed a model for medical records (Shinchi's Medical Record (SMR)) for international disaster relief operations for medical teams from our experience of the Honduras Disaster Relief Operation in November 1998. Our medical unit, which was part of the Japan Self-Defense Forces International Disaster Relief Operation, was confronted with many patients who needed immediate medical care. SMR was developed using only one sheet of paper including the medical record, laboratory data sheet, and prescribed drug sheet. SMR also registers urgency classes and the primary diagnosis. SMR is simple, inexpensive, and easy to prepare for many patients. PMID- 12636138 TI - Long-term hematological effects in Special Forces trainees. AB - The present study assesses the impact of "sports anemia" on screening tests performed on participants in an endurance-training program. Medical charts of 48 naval Special Forces trainees and 48 submarine trainees were reviewed. We compared the results of red blood cell count, hemoglobin concentration, hematocrit (Hct), and mean corpuscular volume performed at recruitment and after 2 years. No differences were noted between baseline values. The Hct and red blood cell count decreased significantly in the Special Forces trainees after 2 years of training (p = 0.002 and p = 0.05, respectively), remaining unchanged in the submariners. Hct decreased by 2 +/- 4.14%, whereas red blood cell count decreased by 0.13 +/- 0.44 M/microL. A reduction in Hct may be expected in Special Forces trainees engaged in long-term endurance training. Because these lower values were not seen in an otherwise identical group of submarine trainees, they could be attributed to the endurance training. PMID- 12636140 TI - Primary squamous cell carcinoma of the thyroid gland. AB - A case of primary squamous cell carcinoma of the thyroid is presented. Squamous epithelium is normally not present in the thyroid gland, and this tumor is exceedingly rare. Proper workup is described, including the differentiation from metastatic disease. The case reported is of a 64-year-old woman who presented with an asymptomatic nodule and underwent thyroidectomy. The final pathologic diagnosis, confirmed using immunohistochemistry for cytokeratin and transmission electron microscopy, revealed the unusual tumor. Squamous cell carcinoma of the thyroid is highly aggressive and behaves clinically like anaplastic thyroid carcinoma; the prognosis is poor. The patient described developed local recurrence and expired 7 months after resection from local invasion and airway obstruction. Unfortunately, this is the typical outcome. Chemoradiotherapy is generally not helpful, and complete resection remains the best chance for cure. PMID- 12636141 TI - Disease and nonbattle injury patterns: afloat data from the U.S. Fifth Fleet (2000-2001). AB - Disease and nonbattle injury (DNBI) patterns were analyzed for reports from the Commander Fifth Fleet for 2000-2001 covering 217,972 person-weeks. The overall DNBI rate was 4.38 visits per 100 person-weeks with the largest subcategory being "other medical/surgical" conditions (1.36 visits/100 person-weeks and over 30% of the initial patient visits). This was followed by dermatological (0.89 visits/100 person-weeks) and respiratory conditions (0.65 visits/100 person-weeks). Collapsing total injuries would have created the third largest incidence category. The mean DNBI rate for cruisers/destroyers/ frigates (4.23 visits/100 person-weeks; SD, 2.64 visits) was not significantly different from aircraft carriers (4.76 visits/100 person-weeks; SD, 1.60, t(df = 207) = -0.91, p = 0.363) but was statistically different from supply ships (8.93 visits/100 person-weeks; SD, 2.44, t(df = 191) = -6.23, p = 0.000) and amphibious support ships (8.07 visits/100 person-weeks; SD, 3.99, t(df = 190) = -4.72, p = 0.000). These results are compared with historical data from land-based units, and the limitations in shipboard DNBI reporting are discussed. PMID- 12636142 TI - Sentinel cases of community-acquired methicillin-resistant Staphylococcus aureus onboard a naval ship. AB - Methicillin-resistant Staphylococcus aureus (MRSA) is emerging as a community acquired organism. A number of recent reports have documented its involvement in a variety of infections in which no risk factors for nosocomial transmission are present. This report presents the initial cases of a MRSA outbreak on a U.S. Navy ship. Each patient failed traditional antibiotic therapy and one required hospitalization. Their presentations evolved simultaneously and proved to be sentinel cases of an outbreak of cutaneous MRSA infections. The events of this outbreak emphasize the growing need to consider the prevalence of resistant organisms in outpatient settings, as well as the impact that infections from resistant organisms might have on the combat readiness of a military unit. Recommendations addressing infection-control guidelines for MRSA within close quarter environments of healthy adults, such as military units, need to be developed and existing infection-control measures need to be regularly emphasized. PMID- 12636143 TI - Teen obesity addressed in the military. AB - OBJECTIVE: The purpose of this study was to trial a teen weight loss program at the largest U.S. Air Force Medical Center to find its effect in reducing the serious problem of teen obesity in San Antonio, Texas, at Lackland Air Force Base. A successful program, developed by physicians, was examined. The program, SHAPEDOWN, selected because it was the first program for teen obesity ever developed and has been successful for over 20 years, used a behavioral modification approach, which required parental involvement. The five students and five parents graduating from the program did reduce their body mass index but not to the program specifications of 1/2 to 1 lb/week. As recommended for success, the program was supposed to include weekly consecutive meetings, pretesting families to note specific family problem areas, and purchasing the training video for the teacher. However, financial and other constraints limited following the program design, and these steps were not taken. Nurturing skills for the parents were taught successfully, and parents expressed future application of these skills for both future weight loss efforts with their teens and for their improved parent/teen relationship. PMID- 12636144 TI - Motor axonal injury and amyotrophic lateral sclerosis: risk assessment using a reverse probability analysis technique. AB - A relationship between antecedent trauma and amyotrophic lateral sclerosis (ALS) has been clinically suspected for over a century. Nine cases of ALS that occurred over a 14-year period in young U.S. men (age range, 28-43 years) following trauma that produced motor axonal injury that antedated (range, 5-42 months) the onset of ALS have been reported by a single neurologist. After estimating the number of cases of ALS in 28- to 43-year-old American men that would be expected to occur over a 14-year period, a reverse probability analysis technique was used to determine the expected incidence rates of motor axonal injury in a ideally matched control population that would give probabilities of 0.05, 0.01, and 0.001 of observing nine or more instances of motor axonal injury occurring during the previous 5 to 42 months. This analysis was repeated assuming that this one neurologist had only reported 50 and 10% of all instances of ALS that occurred during this 14-year period in American men ages 28 to 43 years who had experienced trauma that produced motor axonal injury that occurred 5 to 42 months before the onset of ALS. Reverse probability analysis is a technique that can be used in performing risk assessments. PMID- 12636145 TI - Suicide in the Italian military environment (1986-1998). AB - OBJECTIVE: The aim of our study was to provide data about suicide and attempted suicide in the Italian military environment from 1986 to 1998. METHODS: Italian data were gathered from the clinical documentation archived by the Epidemiological Observatory. These data were collected in a Case Report Form and subsequently sorted into the statistical program database, SPSS, and then processed. RESULTS: From 1986 to 1998, there were 122 suicides and 136 attempted suicides. The most frequently involved ranks were military troops. Subject age range varied from 17 to 60 years. The most frequent ages for both suicides and attempted suicides were 19, 20, 21, and 22 years old. DISCUSSION: The authors assume that the lower predominance of suicide in the military population is mainly attributable to the following factors: screening procedures of military personnel aim to exclude mentally disturbed and the military service provides a young individual with the possibility to belong and to identify himself with a group. PMID- 12636146 TI - Serum dioxin and psychological functioning in U.S. Air Force veterans of the Vietnam War. AB - Using the Minnesota Multiphasic Personality Inventory and the Millon Clinical Multiaxial Inventory, we assessed the psychological functioning of U.S. Air Force veterans exposed to Agent Orange and its contaminant, 2,3,7,8-tetrachlodibenzo-p dioxin (dioxin), during the Vietnam War. Index subjects were veterans of Operation Ranch Hand (N = 1,109). Comparisons (N = 1,493) were U.S. Air Force veterans not involved with spraying herbicides. We found few consistent psychological abnormalities associated with serum dioxin levels. Ranch Hand veterans with higher dioxin levels showed some difficulties in anxiety, somatization, depression, and a denial of psychological factors. However, those with background levels also showed indications of emotional distress, primarily in emotional numbing and lability; a guarded, suspicious, and withdrawn style of relating to others; and unusual thoughts or behaviors. PMID- 12636147 TI - Timing and completeness of routine testing for antibodies to human immunodeficiency virus type 1 among active duty members of the U.S. Armed Forces. AB - Since 1985, the U.S. Armed Forces have conducted routine testing of personnel for antibodies to human immunodeficiency virus type 1. Sera that remain after testing are archived in the Department of Defense serum repository for potential uses in medical surveillance, clinical, and research activities. The goal of this study was to document the timing and completeness of routine human immunodeficiency virus type 1 testing among active duty military personnel and to identify factors associated with more recent testing. Demographic characteristics for active duty personnel were obtained from the Defense Medical Surveillance System. Of all individuals on active duty on August 31, 2001 (N = 1,370,367), 98.6% had been tested. Older personnel, those with postgraduate degrees, and married personnel had the longest time intervals since their most recent tests. The results of this analysis may inform the planning and conduct of human immunodeficiency virus type 1 prevention programs as well as deployment-related or other surveillance activities. PMID- 12636148 TI - Training incidents in armored vehicles in the Singapore Armed Forces. AB - Training in armored vehicles presents occupational hazards. Since the inception of armored units in the Singapore Armed Forces in 1969, there has been no scientific study of the demographics of the trauma patterns. A review of existing literature also indicated a paucity of data on this subject. This article qualified and quantified the proportion of trauma during peacetime armored vehicle training during a 5-year period. Most of the 100 documented incidences of injuries occurred on the head and body limbs. Many injuries were caused by minor lacerations, abrasions, and crush injuries. Although fractures accounted for 24%, 65% of the total injury count could be classified as "minor." A diurnal pattern of injuries was noted in the study. The injury patterns could be used to analyze morbidity and mortality trends and facilitate subsequent evaluation of efficacy of affirmative action. New challenges facing further research in training injuries were also discussed. PMID- 12636149 TI - Identifying clinical supply requirements for Marine Corps dental capability. AB - The Naval Health Research Center developed and used a systematic process to review Marine Corps dental supply requirements. The approach (1) identified the dental procedures required to treat patients with dental conditions and (2) determined the consumable supplies and equipment needed to complete each procedure. By establishing the clinical requirement for each item pushed forward, the Naval Health Research Center model significantly reduced weight, cubic volume, and cost while enhancing far-forward dental capability. This effort produced a database that clarifies the relationship among dental conditions, dental procedures, consumables, and equipment when conducting authorized dental allowance list reviews, determining training requirements, and configuring supplies for future deployments. PMID- 12636150 TI - Fuzzy modeling of skin permeability coefficients. AB - PURPOSE: The purpose of this work was to determine whether a new modeling methodology using fuzzy logic c an predict skin permeability coefficients that are given compound descriptors that have been proven to affect percutaneous penetration. METHOD: Three fuzzy inference models were developed using subtractive clustering to define natural structures within the data and assign subsequent rules. The numeric parameters describing the rules were refined through the use of an Adaptive Neural Fuzzy Inference System implemented in MatLab. Each model was evaluated using the entire data set. Then predicted outputs were compared to the published experimental data. RESULTS: All databases produced fuzzy inference models that successfully predicted skin permeability coefficients, with correlation coefficients ranging from 0.83 to 0.97. The lowest correlation coefficient resulted from a model using log octanol/water partition coefficient and molecular weight as inputs with two input membership functions evaluated by two fuzzy rules. T he correlation coefficient of 0.97 occurred when log octanol/water partition coefficient and hydrogen bond donor activity were used as inputs with three input membership functions evaluated by three fuzzy rules. CONCLUSIONS: Fuzzy rule-based models are a realistic and promising tool that can be used to successfully model and predict skin permeability coefficients as well as or better than previous algorithms with fewer inputs. PMID- 12636151 TI - Analysis of the initial burst of drug release coupled with polymer surface degradation. AB - PURPOSE: Local pH effect on the release of a model pH-inert hydrophobic drug coupled with polymer degradation is described at the induction phase of biodegradable polymer erosion for better understanding the nature of initial burst of a drug. METHODS: Using a novel approach with time-of-flight secondary ion mass spectrometry. both surface concentration of Ph3N and degradation kinetics of PLLA are simultaneously and independently determined from a model Ph3N/PLLA (20:80 wt%) blend matrix (t approximately 0.4 microm on 1.0 cm2). In vitro hydrolysis of the model blend matrix is investigated for short-term periods (<24 h) at physiologic pH and temperature and compared to basic pH. RESULTS: The rate of PLLA degradation is accelerated by a factor of approximately 3 when using basic pH in vitro, but the rate of Ph3N accumulation at the surface is accelerated by a factor of approximately 6. CONCLUSIONS: A new quantitative method has been developed to examine the earliest stages of polymer degradation and drug release. It was applied to a model system that could not be examined by traditional in vitro methods. For the model system studied the release of a low molecular weight hydrophobic drug at the induction phase of polymer erosion is related to but not singularly dependent on degradation kinetics. PMID- 12636152 TI - Poly-L-arginine predominantly increases the paracellular permeability of hydrophilic macromolecules across rabbit nasal epithelium in vitro. AB - PURPOSE: The purpose of this work was to characterize the main transport pathway of hydrophilic macromolecules induced by poly-L-arginine (poly-L-Arg; molecular weight 42.4 kDa) across the excised rabbit nasal epithelium. METHODS: Excised rabbit nasal epithelium was mounted in an Ussing-type chamber for measurement of fluorescein isothiocyanate-labeled dextran (FD-4; molecular weight 4.4 kDa) transport and transepithelial electrical resistance (TEER). The main transport pathway of FD-4 enhanced by poly-L-Arg was evaluated using confocal laser scanning microscopy. Immunolocalization of junction proteins (ZO-1, occludin, and E-cadherin) after treatment with poly-L-Arg was also observed. RESULTS: After apical application of a poly-L-Arg (0.05, 0.5, and 5 mg/mL), the permeability coefficient of FD-4 increased by 1.6-, 2.9-, and 5.2-fold, respectively, compared with the control of 5.2 +/- 1.3 x 10(-7) cm/s. Consistent with the increase in transport, there was a concurrent reduction in TEER. At a concentration of 0.05 mg/mL poly-L-Arg. both FD-4 transport and TEER returned to the control level. A good correlation was obtained between the FD-4 permeability coefficient and 1/TEER. Basolateral application of poly-L-Arg at 5 mg/mL, however, did not increase FD-4 transport. Marked FD-4 fluorescence was located in the paracellular spaces after treatment with apical poly-L-Arg compared with that in the absence of poly-L-Arg. Immunofluorescence of ZO-1, occludin, and E-cadherin in cell-to cell junctions was reduced and distributed into the cytoplasm by apical application of poly-L-Arg, suggesting that poly-L-Arg regulates the junction proteins to enhance paracellular permeability across the nasal epithelium. After pretreatment with either 2,4-dinitrophenol or ouabain, the enhancing effect of apical poly-L-Arg was abolished, indicating the contribution of metabolic energy (cell viability) to the poly-L-Arg-mediated enhancing effect. CONCLUSION: In the nasal epithelium, apical poly-L-Arg appears to increase predominantly the paracellular transport of hydrophilic macromolecules via disorganization of tight and adherens-junction proteins. The regulatory mechanism of the poly-L-Arg effect is likely to be dependent on energy-requiring cellular processes. PMID- 12636154 TI - Modulation of digoxin transport across Caco-2 cell monolayers by citrus fruit juices: lime, lemon, grapefruit, and pummelo. AB - PURPOSE: To evaluate the effects of fresh lime, lemon, grapefruit, and pummelo juices on the transport of digoxin, a P-glycoprotein (P-gp) substrate, in Caco-2 cell monolayers. METHODS: Bidirectional [3H]-digoxin fluxes across confluent Caco 2 cell monolayers were determined in 0-50% fruit juices at pH 7.4. Verapamil HCl (100 microM) served as positive control. Juice toxicity was evaluated by the 3 (4,5 dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay. RESULTS: Apical to-basal (A-to-B) digoxin flux was enhanced by 50% fruit juice in the order of lemon > lime > pummelo > grapefruit. The four fruit juices could be divided into two groups based on their effects on transepithelial electrical resistance (TEER), viability, and digoxin transport activity of the Caco-2 cells. Grapefruit and pummelo juices produced similar digoxin transport profiles that were characteristic of those observed with P-gp inhibitors. Both juices decreased net digoxin efflux by 1.2 U per 10% increase in juice concentration and had a propensity to increase cellular TEER at high concentrations (>30%). However, cellular TEER and viability decreased with increasing concentration of lime and lemon juices. Both juices also produced similar digoxin transport profiles, the A to-B and B-to-A digoxin Papp increasing with increasing juice concentration above 5%. Net digoxin efflux was 30% of control value and relatively independent of juice concentration. These results paralleled the groupings of the four fruits according to their prominent flavonoid pattern and taxonomy. CONCLUSION: The effects of lime, lemon, grapefruit, and pummelo juices on the TEER, viability, and digoxin transport activity of the Caco-2 cells appeared to be dependent on the dominant flavonoid pattern and taxonomy of the citrus fruits. PMID- 12636153 TI - Isolation and characterization of Caco-2 subclones expressing high levels of multidrug resistance protein efflux transporter. AB - PURPOSE: The purpose of this study was to isolate Caco-2 subclones that express high levels of multidrug resistance protein (MDR1) and to characterize their kinetics and affinity parameters for MDR1 substrate/inhibitors. METHODS: The subclones were selected by a dilution cloning technique. The polarized efflux of [3H]-vinblastine across subclone cell monolayers was quantified by measuring the apparent permeability coefficients (Papp) of [3H]-vinblastine in the basolateral (BL)-to-apical (AP) direction and in the AP-to-BL direction (Papp BL-to-AP/Papp AP-to-BL) across the cell monolayers. The expression of MDR1 in the Caco-2 subclones compared with the parental Caco-2 cells was confirmed by Western blotting analysis. The kinetics parameters (Km, Vmax) of [3H]-vinblastine and the inhibitory constants (KI) of several known MDR1 substrates/inhibitors on the transport of [3H]-digoxin determined in the parental Caco-2 cells and Caco-2 subclones were also compared. RESULTS: Three subclones (#1, #20, #21) were selected based on their polarized efflux of [3H]-vinblastine. The Papp BL-to AP/Papp AP-to-BL ratios for #1, #20, and #21 were 110, 140, and 112, respectively, and were about 6-fold higher than the ratio observed for the parental Caco-2 cells. In the presence of GF-120918 (2 microM), a known MDR1 specific inhibitor, the Papp BL-to-AP/Papp AP-to-BL ratios were significantly decreased, suggesting that these cells were overexpressing MDRI. The Km values observed for vinblastine in the Caco-2 subclones were nearly identical to the value observed in the parental Caco-2 cells. In contrast, the Vmax values observed in the subclones were approximate 26-69% higher. The KI values observed for various known MDR1 substrates/inhibitors on [3H]-digoxin transport were nearly identical to those in the parental Caco-2 cells and Caco-2 subclones. The high functional efflux activities of these subclones were stable up to 6 months. CONCLUSIONS: Subclones #1, #20, #21 express high levels of MDR1. These Caco-2 subclones may be useful models for profiling drugs for their MDR1 substrate activity and for establishing structure-transport relationships for this efflux transporter. PMID- 12636155 TI - Physicochemical properties and transport of steroids across Caco-2 cells. AB - PURPOSE: The purpose of this work was to study the relevant physicochemical properties for the absorption of steroids. METHODS: Various physicochemical properties of steroids were calculated (molecular weight, ClogP, static polar surface area [PSA], etc.). Within this series of steroids, different pharmacological groups were defined. Based on the outcome of this survey, steroids were selected for the Caco-2 permeability study. The apparent permeability coefficients (Papp) were related to the calculated and measured physicochemical properties. RESULTS: Between the defined groups of steroids, ClogP was the most discriminative descriptor. The steroids were well transported over the cell monolayers and the Papp was independent of the concentration and the transport direction. No relationship was found with the PSA; however, the Papp showed a weak inverse correlation with ClogP. CONCLUSIONS: The molecular descriptors and Papp values showed that all steroids are well transported. The small differences in the Papp values showed a weak inverse correlation with ClogP: the hydrophilic steroids (ClogP approximately 0-2) tend to diffuse faster over the cell monolayers compared with the more hydrophobic steroids (ClogP approximately 5). The relationship with ClogP suggests that partitioning of steroids between the biologic membrane and the surrounding aqueous phase is one of the main mechanisms for absorption. PMID- 12636156 TI - N-in-one permeability studies of heterogeneous sets of compounds across Caco-2 cell monolayers. AB - PURPOSE: The purpose of the study was to evaluate several n-in-one cocktails of heterogeneous compounds to increase the throughput of permeability studies across Caco-2 monolayers, to investigate the reliability and applicability of the method, and to develop fast and sensitive analysis for the compounds. Compounds with potential interactions in efflux and/or active transport were chosen. METHODS: Permeability experiments with verapamil, propranolol, midazolam, hydroxyzine, timolol, buspirone, procaine, naproxen, ketoprofen, and antipyrine as single compounds and in cocktails of 5-10 compounds were performed at 50 microM concentration both in the apical-to-basolateral and basolateral-to-apical direction. The compounds were quantified by liquid chromatography-electrospray tandem mass spectrometry (LC-ESI/MS/MS). Toxicity tests were performed to determine cellular damage. RESULTS: The analytical method was sensitive, accurate, and rapid. The individual permeabilities of compounds in cocktails correlated well with permeabilities as single compounds. No significant interactions between the compounds within the mixtures were observed, except for acidic compounds. The studied mixtures did not show any toxicity. CONCLUSIONS: The use of n-in-one cocktails is a suitable method to improve the capacity in routine permeability experiments and higher throughput screening of drug candidates, although potential interactions should always be borne in mind. The use of LC-ESI/MS/MS technology provides an excellent tool in fast and accurate analysis of small amounts of heterogeneous compounds. PMID- 12636157 TI - Dextran-methylprednisolone succinate as a prodrug of methylprednisolone: local immunosuppressive effects in liver after systemic administration to rats. AB - PURPOSE: The purpose of this work was to study the local immunosuppressive effects of systemically administered methylprednisolone (MP) and its prodrug, dextran-methylprednisolone (DMP), in rat livers. METHODS: Single 5 mg/kg (MP equivalent) doses of MP or DMP were injected intravenously to rats, and livers were isolated at different time points (0-72 h; n = 4/time point). Isolated livers were stimulated ex vivo with bacterial lipopolysaccharide, and outlet perfusate and bile samples were analyzed for their concentrations of tumor necrosis factor (TNF)-alpha by enzyme-linked immunosorbent assay. The area under the perfusate TNF-alpha concentration-time curve (AUC) was used as a measure of immune response. Hepatic concentrations of MP and DMP were also measured by high performance liquid chromatography. RESULTS: Both MP and DMP resulted in a decrease in lipopolysaccharide-induced increase in TNF-alpha AUC. MP injection 8 h before liver isolation resulted in a maximum of 50% decrease in TNF-alpha AUC. Compared with MP, the maximum effect of the prodrug (DMP) was both more intense (approximately 80% reduction in TNF-alpha AUC) and delayed (maximum inhibition at 24 h). Overall, the area under the effect (% inhibition of TNF-alpha)-time (%inhibition-h) for DMP (3,680 +/- 406) was approximately four times more than that for the parent drug (846 +/- 114). Whereas the MP concentrations in the liver were not quantifiable after the injection of the parent drug, relatively large concentrations of DMP and regenerated MP were found in the liver of DMP injected rats. CONCLUSIONS: After systemic administration to rats, both MP and DMP exhibit local immunosuppressive effects in the liver. The local effects of the prodrug (DMP), however, appear to be more intense and sustained than those of the parent drug (MP). PMID- 12636158 TI - Anhydride prodrugs for nonsteroidal anti-inflammatory drugs. AB - PURPOSE: The objective of this study was to synthesize anhydride prodrugs for prolong action to shield the carboxylic acid group from irritative effects and to temporary hydrophobize the drug so that it becomes accessible to aqueous media when the anhydride residue is hydrolyzed. METHODS: Ibuprofen, a nonsteroidal anti inflammatory agent, was used as a representative drug for anhydride derivatization. Mixed anhydrides of ibuprofen with fatty acids of different chain length were prepared by reacting acid chloride derivatives with the corresponding acid in the presence of acid acceptor and two-phase reaction. Mixed anhydrides were also prepared by dehydration reaction using acetic anhydride and anhydride interchange of symmetric anhydrides. The analgesic effects of mixed anhydride prodrugs were tested using nonsteroidal anti-inflammatory drug rat paw edema model. In vitro degradation of mixed anhydrides and drug release were monitored by high-performance liquid chromatography. RESULTS: Ibuprofen was bound to aliphatic and aromatic acids via an anhydride bond in high reaction yields (>85%) with high mixed anhydride content (>80%). The mix anhydride was purified by chromatography and stored at 4 degrees C to minimize conversion into the symmetric anhydride. These anhydride derivatives hydrolyzed at different time intervals depending on the hydrophobicity of conjugated acid. In vivo testing of the ibuprofen anhydride derivatives for analgesic effect indicated an extended action of the drug for over 24 h as a function of the fatty acid chain length. CONCLUSION: This study demonstrates the promise of anhydride prodrugs for extending drug action and shielding the carboxylic acid group. PMID- 12636159 TI - Dynamics of endocytosis and exocytosis of poly(D,L-lactide-co-glycolide) nanoparticles in vascular smooth muscle cells. AB - PURPOSE: The purpose of this work was to characterize the process of endocytosis, exocytosis, and intracellular retention of poly (D,L-lactide-co-glycolide) nanoparticles in vitro using human arterial vascular smooth muscle cells (VSMCs). METHODS: Nanoparticles containing bovine serum albumin (BSA) as a model protein and 6-coumarin as a fluorescent marker were formulated by a double emulsion solvent evaporation technique. The endocytosis and exocytosis of nanoparticles in VSMCs were studied using confocal microscopy and their intracellular uptake and retention were determined quantitatively using high-performance liquid chromatography. RESULTS: Cellular uptake of nanoparticles (mean particle size 97 +/- 3 nm) was a concentration-, time-, and energy-dependent endocytic process. Confocal microscopy demonstrated that nanoparticles were internalized rapidly, with nanoparticles seen inside the cells as early as within 1 min after incubation. The nanoparticle uptake increased with incubation time in the presence of nanoparticles in the medium; however, once the extracellular nanoparticle concentration gradient was removed, exocytosis of nanoparticles occurred with about 65% of the internalized fraction undergoing exocytosis in 30 min. Exocytosis of nanoparticles was slower than the exocytosis of a fluid phase marker, Lucifer yellow. Furthermore, the exocytosis of nanoparticles was reduced after the treatment of cells with the combination of sodium azide and deoxyglucose, suggesting that exocytosis of nanopartides is an energy-dependent process. The nanoparticle retention increased with increasing nanoparticle dose in the medium but the effect was relatively less significant with the increase in incubation time. Interestingly, the exocytosis of nanoparticles was almost completely inhibited when the medium was depleted of serum. Further studies suggest that the addition of BSA in the serum free medium with or without platelet derived growth factor (PDGF) induced exocytosis of nanoparticles. The above result suggests that the protein in the medium is either adsorbed onto nanoparticles and/or carried along with nanoparticles inside the cells, which probably interacts with the exocytic pathway and leads to greater exocytosis of nanoparticles. CONCLUSIONS: The study demonstrated that endocytosis and exocytosis of nanoparticles are dynamic and energy-dependent processes. Better understanding of the mechanisms of endocytosis and exocytosis, studies determining the effect of nanoparticle formulation and composition that may affect both the processes, and characterization of intracellular distribution of nanoparticles with surface modifications would be useful in exploring nanoparticles for intracellular delivery of therapeutic agents. PMID- 12636160 TI - Phagocytosis and phagosomal fate of surface-modified microparticles in dendritic cells and macrophages. AB - PURPOSE: We compared cationic, polyamine-coated microparticles (MPs) and anionic, protein-coated MPs with respect to their phagocytosis and phagosomal fate in dendritic cells (DCs) and macrophages (Mphi). METHODS: Polystyrene MPs were surface modified by covalent coupling with fluorescein isothiocyanate-labeled polyamines or proteins. Phagocytosis of MP and the pH of their intracellular microenvironment was assessed in human-derived DCs and Mphi in a fluorescence plate reader. Visualization of MP phagocytosis in DCs was performed by transmission electron microscopy. RESULTS: Phagocytosis of bovine serum albumin coated MPs was low with significant differences between DC and Mphi, whereas phagocytosis of IgG-coated MPs was significantly enhanced in both cell types. Phagocytosis of both particle types resulted in an acidified phagosomal microenvironment (pH 4.6-5.1). In contrast, cationic, polyamine-coated MPs were equally phagocytosed by DCs and Mphi to a high extent and showed lower degrees of acidification (pH 6.0-6.8) in the phagosomal microenvironment. Transmission electron microscopy examination demonstrated all phagocytosed particles to be surrounded by a phagosomal membrane, which was more tightly apposed to the surface of cationic MPs and more loosely to bovine serum albumin-coated MPs. CONCLUSION: Phagocytosis of cationic, polyamine-coated MPs is suggested to lead to diminished phagosomal acidification. Thus, cationic MP are potential carriers that may display beneficial features for the intracellular delivery of immunomodulating therapeutics and their protection against lysosomal degradation. PMID- 12636161 TI - Tumor-cell targeted epiderimal growth factor liposomes loaded with boronated acridine: uptake and processing. AB - PURPOSE: The aim of this work was to investigate the cellular binding and processing of polyethylene glycol-stabilized epidermal growth factor (EGF) liposomes. The liposomes were actively loaded with water-soluble boronated acridine (WSA), primarily developed for boron neutron capture therapy. METHODS: The uptake, internalization, and retention of EGF-liposome conjugates were studied in two cultured monolayer cell-lines, A-431 and U-343, with regard to the nuclide-label on the targeting agent, the carrier, and the load. The subcellular localization of WSA was studied using confocal microscopy. RESULTS: We found that the liposome complex was internalized after specific binding to the EGF receptor. After internalization in the tumor cells, WSA was distributed mainly in the cytoplasm and was shown to have long cellular retention, with 80% of the boron remaining after 48 h. CONCLUSIONS: The long retention of the compound and the cellular boron concentration reached makes these targeted liposomes interesting for further development toward boron neutron capture therapy. PMID- 12636163 TI - The effect of CTAB concentration in cationic PLG microparticles on DNA adsorption and in vivo performance. AB - PURPOSE: Cationic PLG microparticles with adsorbed DNA have previously been shown to efficiently target antigen presenting cells in vivo for generating higher immune responses in comparison to naked DNA. In this study we tried to establish the role of surfactant (CTAB) concentration on the physical behavior of these formulations. METHODS: Cationic PLG microparticle formulations with adsorbed DNA were prepared using a solvent evaporation technique. Formulations with varying CTAB concentrations and a fixed DNA load were prepared. The loading efficiency and 24 h DNA release was evaluated for each formulation. Select formulations were tested in vivo. RESULTS: Higher CTAB concentration correlated with higher DNA binding efficiency on the microparticles and lower in vitro release rates. Surprisingly though, the in vivo performance of formulations with varying CTAB concentration was comparable to one another. CONCLUSIONS: Cationic PLG microparticles with adsorbed DNA, as described here, offer a robust way of enhancing in vivo responses to plasmid DNA. PMID- 12636162 TI - Ligand-specific targeting of microspheres to phagocytes by surface modification with poly(L-lysine)-grafted poly(ethylene glycol) conjugate. AB - PURPOSE: The purpose of this study was to demonstrate specific receptor-mediated targeting of phagocytes by functional surface coatings of microparticles, shielding from nonspecific phagocytosis and allowing ligand-specific interactions via molecular recognition. METHODS: Coatings of the comb polymer poly(L-lysine)-g poly(ethylene glycol) (PLL-g-PEG) were investigated for potential to inhibit 1) nonspecific spreading of human blood-derived macrophages (MOs) and dendritic cells (DCs) on glass and 2) nonspecific phagocytosis of PLL-g-PEG-coated, carboxylated polystyrene (PS) or biodegradable poly(D,L-lactide-co-glycolide) (PLGA) microspheres. Coating was performed by adsorption of positively charged PLL-g-PEG on negatively charged microparticles or plasma-cleaned glass through electrostatic interaction. The feasibility of ligand-specific interactions was tested with a model ligand, RGD, conjugated to PEG chains of PLL-g-PEG to form PLL-g-PEG-RGD and compared with inactive ligand conjugate, PLL-g-PEG-RDG. RESULTS: Coatings with PLL-g-PEG largely impaired the adherence and spreading of MOs and DCs on glass. The repellent character of PLL-g-PEG coatings drastically reduced phagocytosis of coated PS and PLGA microparticles to 10% in presence of serum. With both MOs and DCs, we observed ligand-specific interactions with PLL-g PEG-RGD coatings on glass and PS and PLGA microspheres. Ligand specificity was abolished when using inactive ligand conjugate PLL-g-PEG-RDG, whereas repellency of coating was maintained. CONCLUSIONS: Coatings of PLL-g-PEG-ligand conjugates provide a novel technology for ligand specific targeting of microspheres to MOs and DCs while reducing nonspecific phagocytosis. PMID- 12636164 TI - Lung deposition and pharmacokinetics of cyclosporine after aerosolization in lung transplant patients. AB - PURPOSE: Aerosolized cyclosporine (aCsA) has proven to be an effective therapy for refractory acute and chronic rejection in lung transplant (LTx) patients. The objective of this study is to evaluate the lung deposition and systemic absorption of aCsA after aerosolized cyclosporine administration in LTx patients in the immediate postoperative period. METHOD: Cyclosporine (CsA) was administered intravenously (1.0 mg/kg) to eight LTx patients, and multiple blood samples were collected over 24 h. At least 24 h later, aCsA (300 mg in propylene glycol) was administered to the same patients using nebulization and multiple blood samples were obtained again. Five patients had an additional inhalational gamma scintigraphy study with aCsA and 99MTc-labeled albumin to measure drug deposition. RESULTS: Peak blood concentrations of CsA after aerosol administration ranged from 119-402 ng/ml, and concentrations at 24 h ranged from 9-48 ng/ml. The rate of decline in drug concentration in blood in the apparent elimination phase was notably slower after administration of aCsA than after IV infusion. Terminal disposition half life (t 1/2 lambda(z)) values ranged from 4.1 9.9 h (mean 6.5 h) following IV administration and from 23.1 to 65.2 h (mean 40.7 h) following pulmonary administration, suggesting that drug absorption occurred throughout the 24-h sampling period following pulmonary administration. Deconvolution analysis indicated biphasic absorption of CsA from the lung in all patients, characterized by rapid initial absorption (absorption half-life 0.73 +/ 0.38 h) over the first 4 to 6 h followed by slower, sustained absorption throughout the remainder of the sampling period (absorption half-life 16.2 +/- 13.2 h). The absolute bioavailability of CsA after aerosol administration ranged from 5.4-11.2% (mean 8.2%) of the dose placed in the nebulizer. The total dose delivered to the lung estimated from scintigraphy ranged from 17.8-39.3 mg, and was in approximate agreement with the amount of drug absorbed, estimated using deconvolution. Essentially all drug deposited in the lungs was systemically absorbed. CONCLUSIONS: This study documents that cyclosporine can be effectively delivered by aerosolization to the lung of transplant patients in the early postoperative period. Part of the cyclosporine deposited in the lung is absorbed rapidly into systemic circulation and a portion is absorbed slowly but completely over a prolonged period. PMID- 12636165 TI - Dependence of skin permeability on contact area. AB - PURPOSE: We report that experimentally measured skin permeability to hydrophilic solutes increases with decreasing contact area between the formulation and the skin. Our results suggest that an array of smaller reservoirs should thus be more effective in increasing transdermal drug delivery compared to a large single reservoir of the same total area. METHODS: Experimental assessment of the dependence of skin permeability on reservoir size was performed using two model systems, an array of liquid reservoirs with diameters in the range of 2 mm to 6 mm and an array of gel disk reservoirs with diameters in the range of 3 mm to 16 mm. Full thickness pig skin was used as an experimental model. Two molecules, sodium lauryl sulfate (SLS) and oleic acid, were used as model penetration enhancers. RESULTS. Mannitol transport per unit area into and across the skin increased with a decrease in the contact area between the skin and the formulation. Mannitol permeability increased approximately 6-fold with a decrease in the reservoir size from 16 mm to 3 mm in presence of 0.5% SLS in PBS (phosphate buffered saline) as a permeability enhancer. Similar results were obtained when oleic acid was used as an enhancer. CONCLUSIONS: To explain the observed dependence of transdermal transport on contact area a simple mathematical model based on skin geometry in the reservoir was developed. The model predicts a lateral strain in the skin due to preferential swelling of skin upon penetration of water. We propose that this lateral strain is responsible for the increased skin permeability at lower reservoir sizes. PMID- 12636167 TI - Transdermal penetration of vasoconstrictors--present understanding and assessment of the human epidermal flux and retention of free bases and ion-pairs. AB - PURPOSE: As reductions in dermal clearance increase the residence time of solutes in the skin and underlying tissues we compared the topical penetration of potentially useful vasoconstrictors (VCs) through human epidermis as both free bases and ion-pairs with salicylic acid (SA). METHODS: We determined the in vitro epidermal flux of ephedrine, naphazoline, oxymetazoline, phenylephrine, and xylometazoline applied as saturated solutions in propylene glycol:water (1:1) and of ephedrine, naphazoline and tetrahydrozoline as 10% solutions of 1:1 molar ratio ion-pairs with SA in liquid paraffin. RESULTS: As free bases, ephedrine had the highest maximal flux, Jmax = 77.4 +/- 11.7 microg/cm2/h, being 4-fold higher than tetrahydrozoline and xylometazoline, 6-fold higher than phenylephrine, 10 fold higher than naphazoline and 100-fold higher than oxymetazoline. Stepwise regression of solute physicochemical properties identified melting point as the most significant predictor of flux. As ion-pairs with SA, ephedrine and naphazoline had similar fluxes (11.5 +/- 2.3 and 12.0 +/- 1.6 microg/cm2/h respectively), whereas tetrahydrozoline was approximately 3-fold slower. Corresponding fluxes of SA from the ion-pairs were 18.6 +/- 0.6, 7.8+/- 0.8 and 1.1 +/- 0.1 respectively. Transdermal transport of VC's is discussed. CONCLUSIONS: Epidermal retention of VCs and SA did not correspond to their molar ratio on application and confirmed that following partitioning into the stratum corneum, ion-pairs separate and further penetration is governed by individual solute characteristics. PMID- 12636166 TI - Novel microemulsion enhancer formulation for simultaneous transdermal delivery of hydrophilic and hydrophobic drugs. AB - PURPOSE: Microemulsion (ME) systems allow for the microscopic co-incorporation of aqueous and organic phase liquids. In this study, the phase diagrams of four novel ME systems were characterized. METHODS: Water and IPM composed the aqueous and organic phases respectively, whereas Tween 80 served as a nonionic surfactant. Transdermal enhancers such as n-methyl pyrrolidone (NMP) and oleyl alcohol were incorporated into all systems without disruption of the stable emulsion. RESULTS: A comparison of a W/O ME with an O/W ME of the same system for lidocaine delivery indicated that the O/W ME provides significantly greater flux (p < 0.025). The water phase was found to be a crucial component for flux of hydrophobic drugs (lidocaine free base, estradiol) as well as hydrophilic drugs (lidocaine HCl, diltiazem HCl). Furthermore, the simultaneous delivery of both a hydrophilic drug and a hydrophobic drug from the ME system is indistinguishable from either drug alone. Enhancement of drug permeability from the O/W ME system was 17-fold for lidocaine free base, 30-fold for lidocaine HCl, 58-fold for estradiol, and 520-fold for diltiazem HCl. CONCLUSIONS: The novel microemulsion systems in this study potentially offers many beneficial characteristics for transdermal drug delivery. PMID- 12636168 TI - A novel in-vitro technique for studying percutaneous permeation with a membrane coated fiber and gas chromatography/mass spectrometry: part I. Performances of the technique and determination of the permeation rates and partition coefficients of chemical mixtures. AB - PURPOSE: To develop a novel in-vitro technique for rapid assessment of percutaneous absorption of chemical mixtures. METHODS: A silastic membrane was coated on to a fiber to be used as a permeation membrane. The membrane-coated fiber was immersed in the donor phase to partition the compounds into the membrane. At a given partition time, the membrane-coated fiber was transferred into a GC injector to evaporate the partitioned compounds for quantitative and qualitative analyses. RESULTS: This technique was developed and demonstrated to study the percutaneous permeation of a complex mixture consisting of 30 compounds. Each compound permeated into the membrane was identified and quantified with GC/MS. The standard deviation was less than 10% in 12 repeated permeation experiments. The partition coefficients and permeation rates in static and stirred donor solution were obtained for each compound. The partition coefficients measured by this technique were well correlated (R2 = 0.93) with the reported octanol/water partition coefficients. CONCLUSIONS: This technique can be used to study the percutaneous permeation of chemical mixtures. No expensive radiolabeled chemicals are required. Each compound permeated into the membrane can be identified and quantified. The initial permeation rate and equilibrium time can be obtained for each compound, which could serve as characteristic parameters regarding the skin permeability of the compound. PMID- 12636169 TI - The effect of additives on the crystallization of cefazolin sodium during freeze drying. AB - PURPOSE: To monitor the phase transitions during freeze-drying of cefazolin sodium (I) as a function of process and formulation variables. METHODS: Aqueous solutions of I were frozen under controlled conditions in the sample chamber of a variable temperature X-ray powder diffractometer (XRD). The instrument was modified so that the chamber could be evacuated and the samples dried under reduced pressures. Thus, the entire freeze-drying process was carried out in the XRD holder with real time monitoring of the phase transitions during the different stages of freeze-drying. RESULTS: When aqueous solutions of cefazolin sodium (10% w/w) were cooled to -40 degrees C, the XRD pattern revealed only the crystallization of ice. Annealing the frozen sample led to the crystallization of I as the pentahydrate. Differential scanning calorimetry revealed that the presence of isopropyl alcohol (IPA) (5% w/w) led to a decrease in the Tg', the glass transition temperature of the system, and lowered the temperature of crystallization. The crystallization was studied at -8 and at -15 degrees C in the XRD, and, as expected, more rapid crystallization was observed at the higher temperature. Primary drying at -8 degrees C led to the dehydration of the pentahydrate, resulting in a poorly crystalline product. Again, XRD permitted real time monitoring of the decrease in intensities of some characteristic peaks of the pentahydrate. The in situ XRD technique also enabled us to study the effects of processing conditions (different primary and secondary drying temperatures) and crystalline bulking agents on the solid-state of I in the lyophile. When I was lyophilized using mannitol or glycine as an additive, without an annealing step, the drug was X-ray amorphous although the additive crystallized. When annealed and freeze-dried, I remained crystalline in the presence of glycine but not in the presence of mannitol. CONCLUSIONS: The in situ XRD technique has enabled us to characterize the phase transitions during freeze drying of cefazolin sodium in multicomponent systems. PMID- 12636170 TI - Relative reactivity of dihydropyridine derivatives to electrogenerated superoxide ion in DMSO solutions: a voltammetric approach. AB - PURPOSE: To evaluate the reaction of a large series of pharmacologically significant 1,4-dihydropyridine (1,4-DHP) compounds with superoxide (O2.-) in dimethylsulfoxide using differential pulse voltammetry and controlled potential electrolysis. METHODS: Differential pulse voltammetry was used to track the consumption of O2.-, and controlled potential electrolysis was used to electrogenerate O2.-. RESULTS: With the addition of 1,4-DHP, the oxidation peak current of O2.- decreased concentration dependently, suggesting that 1,4-DHP reacts with O2.-, that is, 1,4-DHP scavenges O2.- in dimethylsulfoxide. CONCLUSIONS: very easy and direct voltammetric procedure to study the relative reactivity of different 1,4-DHP with O2.- is proposed. Using the proposed method we have found that all commercial 1,4-DHP reacts with O2.-. The following order of rates was obtained: felodipine > or = vitamin E > isradipine > nimodipine > furnidipine > nitrendipine > nisoldipine > nifedipine. Furthermore, it was demonstrated that the hydrogen at the N-position of 1,4-DHP compounds could be released as a proton in the presence of O2.-, thus the electrogenerated O2.- worked as a proton acceptor to 1,4-DHP. PMID- 12636171 TI - Sterically stabilized phospholipid mixed micelles: in vitro evaluation as a novel carrier for water-insoluble drugs. AB - PURPOSE: Sterically stabilized phospholipid micelles (SSMs) composed of poly(ethylene glycol-2000)-grafted distearoyl phosphatidylethanolamine (PEG(2000) DSPE) are new and promising lipid-based carriers for water-insoluble drugs. This study investigates and compares sterically stabilized mixed micelles (SSMM), composed of (PEG(2000)-DSPE) plus egg-phosphatidylcholine, with SSM as a novel delivery system for improved solubilization of water-insoluble drugs using paclitaxel as a model. METHODS: Paclitaxel was solubilized in SSM (P-SSM) and SSMM (P-SSMM) by coprecipitation and rehydration with isotonic 0.01M HEPES buffer, pH 7.4. After separation of excess drug by centrifugation, mean particle size and morphology of particles in the supernatant were determined by quasi elastic light scattering and transmission electron microscopy. The solubilization potentials of SSMM and SSM for paclitaxel were determined by reverse phase high pressure liquid chromatography (RP-HPLC). Cytotoxic activity of paclitaxel in SSMM. SSM, and dimethyl sulfoxide (10% DMSO) was determined against human breast cancer cells (MCF-7). RESULTS: Mean hydrodynamic diameter of P-SSMM and P-SSM were 13.1 +/- 1.1 nm and 15 +/- 1 nm (n = 3), respectively. SSMM solubilized 1.5 times more paclitaxel than SSM for the same total lipid concentration. Solubilized paclitaxel amount increased linearly with an increase in lipid concentration. A therapeutically relevant lipid concentration (15 mM) of SSMM solubilized 1,321 +/- 48 microg/ml of paclitaxel. Paclitaxel in the absence of sufficient SSM aggregated to form lipid-coated crystals. P-SSMM, P-SSM. and paclitaxel in DMSO had comparable cytotoxic activities against MCF-7 cells. CONCLUSIONS: SSMM showed increased solubilization potential compared with SSM while retaining all of its own advantages. Therefore, it can be used as an improved lipid-based carrier for water-insoluble drugs. PMID- 12636172 TI - Characterization of methacrylated inulin hydrogels designed for colon targeting: in vitro release of BSA. AB - PURPOSE: To characterize methacrylated inulin hydrogels with respect to their release properties. METHODS: Proteins (bovine serum albumin or lysozyme) were used as model drugs and were loaded during or after hydrogel formation. Parameters such as the drug loading method, the molecular weight of the proteins, the initial drug loading concentration, the hydrogel feed composition, degree of substitution, and size of the hydrogel were investigated by determining the release of the model proteins from the hydrogels in a phosphate buffer solution. The biodegradable properties were investigated by studying the release of bovine serum albumin in a solution of inulinase. RESULTS: In vitro protein release from methacrylated hydrogels was influenced by factors such as the drug loading procedure and the molecular weight and loading concentration of the proteins. The feed composition and degree of substitution of inulin seem to be crucial in controlling both the extent and the rate of release. Protein release was clearly enhanced in the presence of inulinase, indicating the biodegradable properties of methacrylated inulin hydrogels. CONCLUSIONS: Several hydrogels show interesting properties with respect to the development of a colon-specific drug delivery system. PMID- 12636173 TI - Microcalorimetric measurement of the interactions between water vapor and amorphous pharmaceutical solids. AB - PURPOSE: Use a microcalorimetric technique to measure the interactions between water vapor and amorphous pharmaceutical solids and describe the relationship between long-term physical stability and the storage relative humidity (RH) at constant temperature. METHODS: A thermal activity monitor was used to characterize interactions of water vapor with spray-dried amorphous sucrose, lactose, raffinose, and sodium indomethacin. Differential scanning calorimetry was used to measure glass transition temperature, Tg. X-ray powder diffraction was used to confirm that the spray-dried samples were amorphous. Scanning electron microscopy was used to examine particle morphology. Specific surface area was determined by BET analysis of nitrogen and krypton adsorption isotherms. RESULTS: The moisture-induced thermal activity traces (MITATs) of the materials in this study exhibit general behavior that helps explain the effect of moisture content on the physical stability of the glassy phase at a given storage temperature. At some RH threshold, RHm, the MITAT exhibits a dramatic increase in the energy of interaction between water vapor and the glass that cannot be explained by a phase or morphology change. Calorimetric data indicate that water vapor-solid interactions are reversible below RHm; above RHm, energetic hysteresis is observed and water-water interactions predominate. In addition, the MITAT was deconvoluted into sorptive and nonsorptive components, making it possible to assign the observed heat flow to unique thermal events. Samples stored at a RH just below RHm for more than 2 months show no evidence of morphology or phase change. In addition, the MITAT can be deconvoluted into sorptive and nonsorptive components by using a twin-calorimeter arrangement. This analysis provides specificity to the microcalorimetric analysis and helps explain the nature of the physical changes that occur during the hydration glassy phase. CONCLUSIONS: The MITAT is a useful tool to determine the onset of moisture induced physical instability of glassy pharmaceuticals and may find a broad application to determine appropriate storage conditions to ensure long-term physical stability. PMID- 12636175 TI - Gene expression profiles of ABC transporters and cytochrome P450 3A in Caco-2 and human colorectal cancer cell lines. AB - PURPOSE: The mRNA levels of MDR1 (P-glycoprotein), multidrug resistance associated proteins (MRP1, MRP2), cytochrome P450 3A (CYP3A) and villin in human colorectal cell lines (HCT-15, LoVo DLD-1, HCT-116 and SW620) were quantitatively compared with those in Caco-2 cells. METHODS: The mRNA levels were determined by real time quantitative polymerase chain reaction and expressed as the relative concen trations of MDR1 mRNA to glyceraldehyde-3-phosphate dehydro genase (GAPDH) mRNA. RESULTSl MDR1 mRNA was expressed in HCT-15 LoVo and DLD-1 cells at similar or lower level to Caco-2. The expression of MRP mRNA in the cell lines tested was comparable with Caco-2. MRP. mRNA was detected only in HCT-116 and SW620 at significantly lower level than Caco-2. CYP3A mRNA was detected in HCT-15 LoVo, DLD-1 and SW620 at similar level to Caco-2. CONCLUSIONS: HCT-15 LoVo and DLD-1 cells express proteins important for regulating the intestinal absorption of drugs, i.e., MDR1 MRP1 and CYP3A, whereas HCT-116 and SW620 cells were not acceptable for evaluation of absorption properties of drug candidates PMID- 12636174 TI - Sorting of rat SPNT in renal epithelium is independent of N-glycosylation. AB - PURPOSE: The sodium-dependent, purine-selective nucleoside transporter, SPNT, has a unique steady-state expression pattern in renal epithelial cells. In comparison with the concentrative nucleoside transporter, CNTI, which is confined to the apical membrane, SPNT is expressed predominantly on the apical membrane but with significant expression on the basolateral membrane as well. Alternate surface expression indicates that SPNT likely has different sorting and trafficking mechanisms from CNTI. Because glycosylation has been reported to be essential for apical targeting of other transporters, and SPNT contains three unique glycosylation sites, we examined the importance of glycosylation in sorting of SPNT. Preliminary studies suggested that glycosylation affects surface expression of SPNT but not CNT1. METHODS: All three unique glycosylation sites were mutated alone and in tandem. Wild-type and mutant SPNT, tagged with green fluorescence protein, were stably transfected into MDCK. Positive clones were assayed for polarized surface expression by immunofluorescence and functional analysis. RESULTS: Mutation at all three sites alone or in tandem resulted in functional proteins. Removal of sites N606 and N625 resulted in proteins of reduced molecular mass. None of the unglycosylated mutants localized differently than wild-type SPNT. CONCLUSION: N-linked glycosylation is not essential for polarized sorting. PMID- 12636176 TI - Development of the liver- and lobe-selective nonviral gene transfer following the instillation of naked plasmid DNA using catheter on the liver surface in mice. AB - PURPOSE: The present study has undertaken the liver- and lobe-selective nonviral gene transfer following the instillation of naked plasmid DNA (pDNA) using catheter on the liver surface in mice. METHODS: The polyethernylon catheter was inserted intraperitoneally through the abdominal wall and was retained on the surface of the liver right and left medial lobes. pDNA was administered through the catheter to the liver right and left medial lobes. RESULTS: The luciferase levels produced in the applied liver lobes at 6 h after liver surface instillation of pDNA were significantly higher than those produced in other liver lobes and other tissues assayed, and ranged from approximately 5 folds higher in other lobes to 20-30 folds higher in other tissues. Following liver surface instillation of pDNA at a time from 2 to 24 h or at a volume from 15 to 60 microl, the gene expressions of the applied liver lobes were always significantly higher than those of other liver lobes and other tissues. CONCLUSION: We have demonstrated the liver- and lobe-selective gene transfection following the instillation of naked pDNA using catheter on the liver surface in mice. PMID- 12636177 TI - Treating Alzheimer's disease with cholinesterase inhibitors: what have we learned so far? PMID- 12636178 TI - The ABC of Alzheimer's disease: ADL and improving day-to-day functioning of patients. AB - Alzheimer's disease (AD) is characterized by deterioration in the ability to perform activities of daily living (ADL) in addition to loss of cognitive function and behavioral changes. This decline in day-to-day functioning is increasingly recognized as a source of considerable social, health, and economic costs. Inability to perform ADL results in growing caregiver burden and may lead to the eventual need for alternative care or nursing home placement. The measurement of ADL, which enables monitoring of the effectiveness of therapeutic interventions, can be performed using a number of inventories including the Progressive Deterioration Scale (PDS), the Disability Assessment for Dementia (DAD), and the Alzheimer Disease Cooperative Study ADL (ADCS/ADL) assessment scale. Clinical studies using these and other scales have indicated that cholinesterase (ChE) inhibitors offer an effective approach to treating the functional decline of AD. Donepezil, rivastigmine, and galantamine have been shown in some studies to prevent or slow decline in ADL over treatment periods of one to two years. For instance, in a 24-week study in subjects with moderate to severe AD, donepezil-treated patients remained stable compared with the placebo treated patients. Rivastigmine has shown improvement or stabilization of PDS scores in patients with mild to moderate disease following 26 weeks of treatment and slowed deterioration in patients with more severe disease. Evidence to date suggests that these agents may not be equally effective at slowing or stabilizing loss in ADL over time and that these differences may reflect differences in pharmacology. In addition to inhibition of acetylcholinesterase (AChE), these compounds have other putative differences in mechanisms of action. Galantamine allosterically modulates the nicotinic receptor and may prevent the loss of ADL. Rivastigmine robustly inhibits butyrylcholinesterase in addition to AChE and therefore acts as a dual ChE inhibitor. Comparative studies evaluating the differential effects of these ChE inhibitors on ADL are awaited. PMID- 12636179 TI - The ABC of Alzheimer's disease: behavioral symptoms and their treatment. AB - Behavioral and psychological symptoms of dementia (BPSD) are a common manifestation of Alzheimer's disease (AD) and other dementia syndromes. Patients experience prominent and multiple symptoms, which are both distressing and a source of considerable social, health, and economic cost. Development of symptoms is in part related to progressive neurodegeneration and cholinergic deficiency in brain regions important in the regulation of behavioral and emotional responses including the cortex, hippocampus, and limbic system. Cholinesterase (ChE) inhibitors offer a mechanism-based approach to therapy to enhance endogenous cholinergic neurotransmission. Studies using ChE inhibitors have demonstrated their clear potential to improve or stabilize existing BPSD. Differences have been noted between selective acetylcholinesterase (AChE) inhibitors (donepezil and galantamine) and dual ChE inhibitors (rivastigmine) in terms of treatment response. While donepezil has shown efficacy in moderate to severe noninstitutionalized AD patients, conflicting results have been obtained in mild to moderate patients and in nursing home patients. Galantamine has been shown to delay the onset of BPSD during a five-month study but has been otherwise poorly studied to-date. Both donepezil and galantamine have not as yet demonstrated efficacy in reducing psychotic symptoms or in reducing levels of concomitant psychotropic medication use. Studies with the dual ChE inhibitor rivastigmine in mild to moderately severe AD and in Lewy body dementia (LBD) have shown improvements in behavioral symptoms including psychosis. Improvements have been maintained over a period of up to two years. In addition, institutionalized patients with severe AD have shown symptomatic benefits with a reduction in the requirement for additional psychotropic drugs following treatment with rivastigmine. The psychotropic properties associated with rivastigmine may in part be mediated through effects on butyrylcholinesterase. Current treatment options are limited for patients with dementia syndromes other than AD. However, data concerning rivastigmine in patients with LBD and preliminary studies in Parkinson's disease dementia and vascular dementia suggest a role for ChE inhibitors across the spectrum of dementia syndromes. Finally, studies that incorporated a delayed start design demonstrate that ChE inhibitors may delay the progression of BPSD. PMID- 12636180 TI - The ABC of Alzheimer's disease: cognitive changes and their management in Alzheimer's disease and related dementias. AB - Cognitive decline, commonly first recognized as memory impairment, is a typical feature of Alzheimer's disease (AD). Neuropathological changes in the cerebral cortex and limbic system lead to deficits in learning, memory, language, and visuospatial skills. The precise nature of cognitive dysfunction reflects the distribution of pathological changes in AD. These will vary along the disease severity continuum and may also depend on where the disease sits in the spectrum of dementia. For example, AD-related disorders such as Lewy body dementia (LBD) and Parkinson's disease dementia (PDD) also show symptoms of cognitive decline and share several pathological features, including degeneration of cortical cholinergic and striatal dopaminergic neurons. In vascular dementia (VaD), there is often an unequal distribution of cognitive deficit, with severe impairment in some functions and relative sparing of others. Cholinesterase (ChE) inhibitors, which help restore acetylcholine levels in the brain, are licensed for the symptomatic treatment of AD and have shown additional benefit in related dementias. Physiological correlates of cholinergic function/dysfunction in the brain include regional cerebral blood flow, glucose metabolism, and cerebrospinal fluid levels of ChE enzymes. These variables represent valuable markers of the clinical efficacy of ChE inhibitors. However, direct assessment of cognitive improvement, stabilization or decline is usually considered the key efficacy parameter in clinical studies of ChE inhibitors in AD and related dementias. Large-scale, placebo-controlled clinical studies of ChE inhibitors have demonstrated efficacy in treating the cognitive impairments associated with AD. Randomized comparative studies of ChE inhibitors are now under way to directly compare symptomatic efficacy and effects on disease progression. Clinical trial data of the cognitive effects of ChE inhibitors in AD, LBD, PDD, and VaD are discussed in detail in this article. The benefits of long-term treatment on symptomatic improvement in cognition and further potential disease-modifying effects are highlighted. PMID- 12636181 TI - Butyrylcholinesterase: an important new target in Alzheimer's disease therapy. AB - Acetylcholinesterase (AChE) predominates in the healthy brain, with butyrylcholinesterase (BuChE) considered to play a minor role in regulating brain acetylcholine (ACh) levels. However, BuChE activity progressively increases in patients with Alzheimer's disease (AD), while AChE activity remains unchanged or declines. Both enzymes therefore represent legitimate therapeutic targets for ameliorating the cholinergic deficit considered to be responsible for the declines in cognitive, behavioral and global functioning characteristic of AD. The two enzymes differ in substrate specificity, kinetics and activity in different brain regions. Experimental evidence from the use of agents with enhanced selectivity for BuChE (cymserine analogues, MF-8622) and the dual inhibitor of both AChE and BuChE, rivastigmine, indicates potential therapeutic benefits of inhibiting both AChE and BuChE in AD and related dementias. Recent evidence suggests that both AChE and BuChE may have roles in the aetiology and progression of AD beyond regulation of synaptic ACh levels. The development of specific BuChE inhibitors and further experience with the dual enzyme inhibitor rivastigmine will improve understanding of the aetiology of AD and should lead to a wider variety of potent treatment options. PMID- 12636182 TI - A clinical overview of cholinesterase inhibitors in Alzheimer's disease. AB - This review provides an overview of the three most widely used cholinesterase (ChE) inhibitors: donepezil, rivastigmine, and galantamine. Differences in pharmacologic profiles will be discussed, and consideration will be given to how such differences may relate to and influence the clinical efficacy and tolerability of the various agents. In addition to providing cognitive benefits in patients with Alzheimer's disease (AD), growing clinical evidence also suggests that ChE inhibitors can produce favorable and clinically relevant effects on neuropsychiatric/behavioral disturbances and activities of daily living. Furthermore, recent data indicate that these agents may be effective at all levels of disease severity and for all rates of disease progression. The clinical utility of ChE inhibitors in a wider spectrum of dementias which share a common cholinergic deficit, such as Lewy body dementia, Parkinson's disease dementia, and vascular dementia, is currently under investigation. Beyond symptomatic relief, data suggest that ChE inhibitors may also slow the underlying disease process. As clinical and research experience with these agents continues to accumulate, the differences in their effects will become more apparent and will help physicians tailor ChE inhibition treatment to the needs of the individual patient. PMID- 12636183 TI - Influence of ventilatory settings and sampling position on measurements of simulated exhaled nitric oxide levels. AB - Chronic lung disease is a common adverse outcome of prematurely born infants and is associated with an early inflammatory response, which persists over weeks. As a consequence, it is possible that exhaled nitric oxide levels might be raised in affected infants. The majority of such infants will be ventilated in the first weeks of birth and thus it is important to determine the influence of mechanical ventilation on nitric oxide levels. As a consequence, our aim was to determine whether, during mechanical ventilation, simulated exhaled nitric oxide levels were influenced by changes in ventilator settings or the sampling catheter position. A lung model was created consisting of a rubber bag inside a 11 jar. An endotracheal tube (ETT) was fixed securely within the neck of the bag. Nitric oxide was delivered into the bag at a constant rate to simulate production and sampled from within the ETT and the bag. The sampled nitric oxide was analysed using a Sievers chemiluminescence analyser. The ETT was attached to a neonatal ventilator and a nitric oxide scavenger placed in the ventilator's inspiratory limb to ensure nitric oxide free gas was delivered. Comparison of different sampling positions revealed that the highest peak nitric oxide level within the ETT was at the tip. Increasing peak inflating pressure and ventilator rate resulted in a decrease in the peak nitric oxide levels. Increasing the inspired oxygen concentration also was associated with a reduction in the peak nitric oxide levels, the effect being more pronounced when larger volume lung models were examined. The results emphasized that the conditions of measurement must be standardized in infants receiving respiratory support, if exhaled nitric oxide results are to be appropriately interpreted. PMID- 12636184 TI - Forced oscillation technique in the sleep apnoea/hypopnoea syndrome: identification of respiratory events and nasal continuous positive airway pressure titration. AB - Sleep breathing disorders (SBD) are related to obstructions resulting from repetitive narrowing and closure of the pharyngeal airway. Their diagnoses and treatment are critically dependent on an accurate identification of and discrimination between types of respiratory events. However, these disorders have been diagnosed using indirect or invasive measurements, which resulted in serious doubts concerning the correct evaluation of breathing events. The forced oscillation technique (FOT) has recently been suggested as a clinical tool able to accurately and non-invasively quantify respiratory obstruction during sleep. The present study investigates the morphology of the impedance signal during different sleep respiratory events and evaluates the ability of impedance measurements in providing adequate nasal continuous positive airway pressure (nCPAP) titration. The results evidenced characteristic patterns in impedance signal morphology that are useful in the identification and classification of abnormal respiratory events. Moreover, significantly higher impedance values were obtained during apnoea and hypopnoea events when compared with normal values (p < 0.01). Studies using impedance measurements to adjust nCPAP showed a significant reduction (p < 0.01) of abnormal respiratory events, and a consequent normalization of the patients. These findings support the use of the FOT as a versatile clinic diagnostic tool helping SBD diagnosis and treatment. PMID- 12636185 TI - Transfer function analysis for clinical evaluation of dynamic cerebral autoregulation--a comparison between spontaneous and respiratory-induced oscillations. AB - Oscillations of arterial blood pressure (ABP) and cerebral blood flow velocity (CBFV) can be used for non-invasive assessment of cerebral autoregulation using transfer function analysis. Either spontaneous oscillations (SPO) around 0.1 Hz or respiratory induced oscillations during deep breathing (DB) at a rate of 6/min have been used so far. We investigated 168 patients with severe carotid stenosis or occlusion to evaluate transfer function analysis and compare the SPO and DB approaches. ABP was assessed non-invasively (Finapres), CBFV was measured in the middle cerebral artery using transcranial Doppler sonography. Transfer function phase (P) and gain (G) were extracted from the respective spectra in a low (0.06 0.12 Hz) and high (0.20-0.30 Hz) frequency range (LF, HF) of SPO and from the 0.1 (LF) and 0.2 (HF) Hz peaks induced by DB. For SPO, significant side-to-side differences and differences between groups of severe and critical stenosis were found for P(LF), while P(HF) did not prove to be a significant parameter. G(LF) showed significant side-to-side differences, while G(HF) additionally differed significantly between severe and critical stenosis and occlusion, respectively. For DB, significant side-to-side differences were found for P(LF, HF). Mainly G(HF) differed significantly between the affected and contralateral sides, while both HF and LF gains showed lower values in groups with a higher degree of stenosis. Correlation between G and P values was generally poor. Using Bland Altman plots a poor inter-method agreement was found mainly for P. Correlations between SPO and DB were higher for G than for P (LF r = 0.64 versus 0.44, HF 0.69 versus 0.28). Analysing reproducibility in 16 patients, only for P(LF, HF) of DB was a highly significant correlation found (Spearman's r up to 0.78). For G(LF, HF) correlations were significant for both SPO and DB with slightly higher r coefficients for SPO. In conclusion, the present study showed that (1) transfer functions P and G represent different information for characterization of dynamic cerebral autoregulation in the frequency domain. (2) Inter-method agreement between DB and SPO is poor for P and moderate for G values. (3) P extracted from DB has a higher reproducibility. (4) The extraction of P and G from the SPO phase spectra is critical and future work on standardizing this process is needed. (5) At present, the DB protocol might be slightly advantageous as a routine diagnostic tool. PMID- 12636186 TI - Physiological interpretation of electrical impedance epigastrography measurements. AB - Measurement of the electrical impedance of the gastric region is carried out with the epigastrograph. This generates and applies alternating current around the abdominal area and measures the potential difference in order to determine the impedance externally, via electrodes. The change of epigastric impedance for a subject, given a meal after fasting, depends on the conductivity of the meal compared to the stomach and surrounding tissues. Typically a conductive meal has conductivity >7 mS cm(-1), non-conductive <2 mS cm(-1) and neutral about 4.5 mS cm(-1). Half-emptying times (T50s) from gastric emptying studies in volunteers using three test meals of 450 ml volume were obtained and found to be shorter than expected from the literature. The meals were a 10% glucose solution and two milk shakes of energy 1,300 kJ and 2,850 kJ, respectively. These electrical impedance epigastrography (EIE) measurements were carried out with scintigraphy. The T50 values of the latter were significantly longer. The direct comparison of the normalized experimental data obtained by both methods led to the concept that EIE measurements are mainly influenced by gastric secretion. Thus the EIE trace of a 'neutral' meal suggests the hypothesis that the volume of the meal is not the significant factor but is influenced by gastric acid secretions. Physiology of the gastric mucosa during the digestion of a meal and intragastric pH values also suggests this. Gastric function studies using EIE measurements may therefore reflect gastric ionic concentration rather than the volume of the contents of the stomach. In turn this could lead to the development of a non-invasive method for the continuous recording of gastric acid secretions. PMID- 12636187 TI - Factors influencing pricking pain threshold using a CO2 laser. AB - The purpose of this study was the evaluation of the best experimental conditions in healthy subjects for the measurement of the minimal thermal energy density E1 which induced pricking pain on the volar surface of the left forearm by means of CO2 laser pulses. E1 was measured on a well-defined area, using laser pulses of different durations and constant power P. The dependence of E1 on the stimulus power P, the size A of the radiated area and the surface temperature T(e) were explored. In the first part of the study, these relations were obtained using a computer program, from the calculated spatio-temporal distribution of the skin temperature during, and following, a laser pulse which caused pricking pain. The second part studied a set of subsequent measurements carried out on a group of five healthy trained subjects and agreed only in part with the calculated data. We found that the measurement error on E(t) was less than 10% with P between 1.5 and 3 W, and A between 0.15 and 0.25 cm2, respectively. The influence of sensitization and adaptation phenomena on the measured data was also explored. We also show a rhythmic annual change of T(e) and E1. PMID- 12636188 TI - Perturbation of blood flow as a mechanism of anti-tumour action of direct current electrotherapy. AB - Anti-tumour effects of direct current electrotherapy are attributed to different mechanisms depending on the electrode configuration and on the parameters of electric current. The effects mostly arise from the electrochemical products of electrolysis. Direct toxicity of these products to tumour tissue is, however, not a plausible explanation for the observed tumour growth retardation in the case when the electrodes are placed into healthy tissue surrounding the tumour and not into the tumour itself. The hypothesis that the anti-tumour effectiveness of electrotherapy could result from disturbed blood flow in tumours was tested by the measurement of changes in blood perfusion and oxygenation in tumours with three different methods (in vivo tissue staining with Patent Blue Violet dye, polarographic oximetry, near-infrared spectroscopy). The effects induced by electrotherapy were evaluated in two experimental tumour models: Sa-1 fibrosarcoma in A/J mice and LPB fibrosarcoma in C57B1/6 mice. We found that perfusion and oxygenation were significantly decreased after electrotherapy. Good agreement between the results of different methods was observed. The effect of electrotherapy on local perfusion of tumours is probably the prevalent mechanism of anti-tumour action for the particular type of electrotherapy used in the study. The importance of this effect should be considered for the optimization of electrotherapy protocols in experimental and clinical trials. The non-invasive technique of near-infrared spectroscopy proved to be a reliable method for detecting perfusion and oxygenation changes in small solid tumours. PMID- 12636189 TI - Adaptive, autoregressive spectral estimation for analysis of electrical signals of gastric origin. AB - The electrical activity of the human stomach, which normally shows a frequency of about 0.05 Hz, may be studied non-invasively by either cutaneous electrogastrography (EGG) or surface magnetogastrography (MGG). Detection of changes in frequency with time may be useful to characterize gastric disorders. The fast Fourier transform (FFT) has been the most commonly used method for the automated spectral analysis of the signals obtained from the EGG or the MGG. We have used an autoregressive (AR) parametric spectrum estimator to analyse simulated signals of gastric electrical activity, and to evaluate the results of human studies using EGG and MGG. In comparison with the FFT, our results showed that the AR spectrum estimator provided more detailed qualitative information about frequency variations of short duration simulated signals than the FFT. In the human studies, the AR estimator was as good as the conventional FFT methods in detecting physiological changes in frequency and in identifying abnormal recordings. We conclude that the AR spectral estimator may provide a better qualitative analysis of frequency variations in small portions of the signal, and is as useful as the FFT to analyse human EGG or MGG studies. PMID- 12636190 TI - Time variation of ammonia, acetone, isoprene and ethanol in breath: a quantitative SIFT-MS study over 30 days. AB - A study of the concentrations of the common breath metabolites ammonia, acetone, isoprene, ethanol and acetaldehyde in the breath of five subjects over a period of 30 days has been carried out. Breath samples were taken and analysed in the early morning on arrival at the laboratory. The real time analyses of three consecutive breath exhalations were carried out using selected ion flow tube mass spectrometry (SIFT-MS) on line to the instrument. Sufficient data were obtained to allow meaningful concentration distributions to be obtained for ammonia, acetone, isoprene and ethanol. These showed that the ammonia, acetone and isoprene concentrations exhibited sensibly normal distributions, with coefficients of variation of typically 0.3. Obvious and statistically significant (p < 0.01) differences are apparent in the mean concentrations of these metabolites between the five individuals. The acetaldehyde concentrations were relatively low and close to the instrument detection limit, and the differences between the mean concentrations of the five subjects were not statistically significant (p = 0.4), so distributions were not obtained. The mean concentrations, in parts per billion (ppb), of each metabolite range amongst the five subjects are as follows: ammonia, 422-2389: acetone, 293-870; isoprene, 55 121; ethanol, 27-153; acetaldehyde, 2-5. There are no obvious patterns in the distributions of these particular metabolites for these individuals, except that the ammonia levels were greatest in the breath of the two oldest subjects. PMID- 12636191 TI - A study of the morphological parameters of cervical squamous epithelium. AB - Electrical impedance spectroscopy is a technique that has been investigated as a potential method for the diagnosis of epithelial carcinomas. Finite element modelling can provide an insight into the patterns of current flow in normal and pathological epithelium and hence aid in the process of probe design optimization. In order to develop a finite element model of the structure of normal and precancerous cervical squamous epithelium, it was first necessary to obtain the mean values and ranges of a number of morphological tissue parameters. The most important parameters in discriminating normal from neoplastic tissue were identified as being cell size and shape distribution, nuclear-to-cytoplasmic volume ratio and volume of extracellular space. A survey of the literature revealed an absence of reliable quantitative data for these parameters. We therefore present the results of our own basic image analysis on normal and pathological tissue sections, which we hope will be of use to other workers wishing to model cervical squamous epithelium, or other similar tissue structures. PMID- 12636192 TI - Dielectric properties of blood: an investigation of haematocrit dependence. AB - We have investigated the haematocrit dependence of the electrical parameters (relative permittivity and conductivity) of blood. The measuring set-up, composed of an impedancemeter (HP 4291 A), an open-ended coaxial line and a temperature controlling set, was designed for dielectric measurements in the 1 MHz to 1 GHz frequency range. Measurements were performed on ex vivo animal (cow and sheep) blood at 37 degrees C. The two dielectric parameters appeared to be strongly dependent on the haematocrit. The permittivity versus frequency decreases then increases when the haematocrit decreases. The conductivity increases in the whole frequency range when the haematocrit decreases. Due to the lack of comparative data on the frequency range explored, we compare the dielectric profiles with those deduced from the Maxwell-Fricke theoretical model. PMID- 12636193 TI - Automated, real-time calibration of the respiratory inductance plethysmograph and its application in newborn infants. AB - Respiratory inductive plethysmography (RIP) is widely used in infants, children and adults. The technique is well accepted as it provides important qualitative information on the pattern of breathing, although its ability to record volume accurately was questioned due to calibration uncertainties. Existing calibration methods require two-position calibration, or patient cooperation in performing various breathing manoeuvres, or prolonged calibration paradigms. The disadvantages from calibration difficulties are even more pronounced in infants. We present a computer system that is capable of performing a single-posture, real time RIP calibration during natural breathing and is suitable for use in newborns. The calibration algorithm is based on interactive, point-by-point calculations of maximal correlation between airflow at the mouth, Vao, and summed differentiated RIP signals. The quantities are calculated interactively at every sample point, and the process continues until stable results are reached and convergence criteria met. A graphic user interface was developed to assist in the rapid implementation and ease of use. Validation schemes were evaluated in 33 newborn infants against actual Vao. Calibration factors were obtained within 21 +/- 11 s with a mean correlation coefficient of 0.97 +/- 0.03. All RIP-derived values were similar to actual airflow signals, with error values ranging from 0.4 +/- 3.0% for respiratory rate to 1.8 +/- 7.3% for tidal volume. Calibration was found to be stable and reliable for up to 3.5 h and in changing sleep states. It is concluded that the new single-posture real-time RIP calibration system is safe and simple to use, and also quick, accurate and stable. The system was found to be suitable for use in newborns during natural breathing while asleep. PMID- 12636194 TI - Quantitative analysis of bone mineral content by x-ray microtomography. AB - A new non-destructive method based on x-ray microtomography (micro-CT) was developed to measure calcium density in bone. X-ray micro-CT was used as a quantitative approach to acquire and reconstruct virtual cross-sections through the sample. Accurate beam-hardening correction was implemented. Grey values in the virtual cross-sections were calibrated as calcium mineral density in bone. From these cross-sections, three-dimensional models were created. Calcium content was calculated directly from images and expressed as percentage per volume and per weight. Calcium mineral density was studied by this method in a unique set of bones isolated from newts (Pleurodeles waltlii Michah) that had travelled into space. A demineralization of 10% was shown as a consequence of sustained micro gravity. PMID- 12636195 TI - Non-invasive flow estimation in an implantable rotary blood pump: a study considering non-pulsatile and pulsatile flows. AB - Non-invasive estimation of flow was investigated in an implantable rotary blood pump (iRBP) with a hydrodynamic bearing. The effects of non-pulsatile and pulsatile flows were studied using in vitro mock loops, and acute (N = 3) and chronic (N = 6) ovine experiments. Using the non-pulsatile and pulsatile mock loops an average flow estimation algorithm was derived from root mean square (RMS) pump impeller speed and RMS input power. These algorithms were programmed into the iRBP controller for subsequent validation in vivo. In the acute experiments, venous return and systemic vascular resistance were adjusted through pharmacological intervention and exsanguination to produce an average range of pump flows from 0.0 to 2.6 l min(-1). Over this range the RMS estimation error was 88 +/- 12 ml, with a linear correlation slope of 0.992 +/- 0.006 (R2 = 0.986 +/- 0.004). In the chronic experiments, animals were monitored daily for up to three months and an average range of flows from 2.8 to 4.8 l min(-1) recorded. A linear correlation between the estimated and measured pump flows yielded a slope of 1.005 +/- 0.006 (R2 = 0.966 +/- 0.004). The RMS estimation error was 120 +/- 11 ml. Using this algorithm it is possible to effectively estimate flow in a rotary blood pump without implanting additional invasive sensors. PMID- 12636196 TI - Increase of acetone and ammonia in urine headspace and breath during ovulation quantified using selected ion flow tube mass spectrometry. AB - Selected ion flow tube mass spectrometry (SIFT-MS) has been used for a detailed study of the daily variations in the acetone and ammonia content of the headspace above urine from a healthy female subject over the course of three separate menstrual cycles. Midstream urine samples were taken every morning prior to any food intake and the headspace subsequently analysed for a number of metabolites. Concurrent with the time of ovulation, a 3-to- 12-fold increase in the level of acetone in the urine headspace was observed. The successive peaks in acetone level and the subsequent return to baseline values were mirrored by similar increases in the ammonia levels, but these were a day out of phase. Interestingly, parallel breath analyses at ovulation showed no great increase in either acetone or ammonia above their normal morning levels, suggesting that these metabolites had been removed from the body during the night by the usual metabolic and physiological processes. The results of this study reveal what may be an important phenomenon at the time of ovulation and illustrate the potential and power of online SIFT-MS analysis in this area of research. PMID- 12636197 TI - Graphical display of variability and inter-relationships of pressure signals in children with traumatic brain injury. AB - A prospective observational study was undertaken to examine time series ICU data of pressure variables (mean arterial pressure (MAP), intracranial pressure (ICP) and cerebral perfusion pressure (CPP)) and relate their variability (SD) to outcome, together with simple graphical displays which could be useful at the ICU bedspace. Forty-three children (aged < 1-15 years) were admitted to the intensive care unit for Regional Neurosurgical Service, Edinburgh, following traumatic brain injury (TBI). The standard deviations from 221,291 validated pressure data measurements (representing three variables) were calculated for the duration of ICP monitoring (and in 48 h epochs from the time of injury). Data were displayed on polygraphs, and several well-defined 'patterns' were described. The standard deviations of MAP, ICP and CPP for the total duration of monitoring were found to be significantly related to survival (p = 0.003, <0.001 and 0.005, respectively), while the SD of ICP alone was strongly related to global recovery (p = 0.008) in the first 48 h post-injury. Patterns in 104 epochs (each of 48 h) were identified. Ninety-two were of the type I (MAP > CPP > ICP) pattern and 12 were of the non-type I pattern. Glasgow Outcome Scale scores at 12 months were significantly related to the dichotomized pattern type (Fisher's exact test p < 0.001 for both alive versus dead and independent versus dependent outcomes). Only one patient with type I pattern died in this series. While variability of ICP during the first 48 h post-injury is predictive of the outcome, the pattern behaviour of three pressure signals gives useful outcome prediction information throughout monitoring. These displays may help interpret some of the plethora of data produced at the bedside. PMID- 12636198 TI - Electrical impedance of muscle during isometric contraction. AB - Non-invasive measurements of the 50 kHz impedance of the anterior forearm show that the resistance and reactance increase under voluntary isometric contraction of the finger flexor muscles. The relationship between impedance and force is nonlinear, dependent on the type of test, the history of prior exercise, and the health status of the subject. Nevertheless, useful dynamic response parameters betaR = deltaR/R0deltaF and betax = deltaX/X0deltaF can be defined, typically a few hundredths of a per cent per newton. Evidence is presented for the view that these effects reflect dominantly physiological as opposed to morphological changes in the muscle. In particular, (a) the impedance changes many milliseconds before the force is generated, (b) betaR and betaX change substantially during a series of repetitions of the same exercise, and (c) the impedance does not return to its original value following relaxation of the muscle. Supporting data are presented for six healthy men and women, with ages ranging from 19 to 70 years. A preliminary study of patients with various neuromuscular diseases was also performed, amongst whom marked quantitative and qualitative contrasts with the healthy group were found. Further research aimed at assessing the clinical potential of such measurements is discussed, as are studies to elucidate the underlying mechanisms for the impedance changes. We propose the name 'dynamic electrical impedance myography' for this new technique. PMID- 12636199 TI - A direct analytical demonstration of the essential equivalence of detrended fluctuation analysis and spectral analysis of RR interval variability. AB - Detrended fluctuation analysis (DFA) of heart rate variability appears to yield improved prognostic power in cardiovascular disease, through calculation of the fractal scaling exponent alpha. We have recently used Taylor series approximations to propose that DFA is closely related to standard spectral analysis measures. We now present a direct analytical approach using integration and explicit expressions for the steps involved in DFA. This demonstrates clearly how DFA corresponds to standard spectral analysis. PMID- 12636200 TI - Autoimmune hepatitis: truly a rare disorder in Taiwan. PMID- 12636201 TI - Characteristics of autoimmune hepatitis in Taiwan: the 11 years' experiences of a medical center. AB - BACKGROUND: Unlike in Western countries, autoimmune hepatitis (AIH) is an infrequent diagnosis in Taiwan. The clinical characteristics of AIH in this area are unclear. The aim of this study was to elucidate the clinical features of AIH in Taiwan. METHODS: All the medical records of in-patients with the diagnosis of chronic hepatitis in our hospital from 1990 to 2001 were reviewed for the possibility of AIH. The clinical features, biochemical data, immunological presentations, treatments and survival of the patients were evaluated. RESULTS: Twenty-two patients (15 females and 7 males) were diagnosed as having AIH within 11 years. The median age at onset was 64 years (range: 17-77 years). Compared with female patients, male patients had older age (p = 0.001), shorter duration from initial presentation of symptoms to diagnosis (p = 0.015), lower serum levels of alkaline phosphatase (ALK-P, p = 0.022) and albumin (p = 0.043). Five (23%) patients presented with cirrhosis upon diagnosis. Compared with non cirrhotic patients, cirrhotic patients had lower serum levels of alanine aminotransferase (p = 0.002), aspartate aminotransferase (p = 0.015), gamma glutamyl transferase (G-GT, p = 0.002), albumin (p = 0.14), white cell counts (p = 0.009) and platelet counts (p = 0.002). Thirteen (59%) patients had concomitant clinico-pathological features of cholestatic liver disease (ALK-P > or = 2 times of upper normal limit or pathologic evidence of cholangiopathy). They had higher serum levels of ALK-P (p < 0.001) and G-GT (p = 0.004) than 9 non-cholestatic patients. There were no significant differences in survival between these groups. The prescribed initial and maintained prednisolone dosages for our patients to control disease activity were 19 +/- 15 mg and 8 +/- 1 mg, respectively, which were lower than those recommended in Western countries. The remission rate to steroid treatment and relapse rate after discontinuing corticosteroids were 87.5% and 50%, respectively. CONCLUSIONS: Compared with Western AIH patients, the AIH patients in Taiwan are older and more likely to develop cholestasis, and need a relatively lower dose of steroid for treatment. Owing to one quarter of the patients already having liver cirrhosis on diagnosis, AIH should be suspected in any Taiwanese patient with cryptogenic hepatitis or cirrhosis. PMID- 12636202 TI - Genistein and tyrphostin AG 556 block the action potential shortening in septic shock. AB - BACKGROUND: We have previously shown that an increase in NO activity activated ATP-sensitive potassium channel (K(ATP)) and shortened action potential duration (APD) in an endotoxic shock model. Because the increase in NO production and the decrease of APD appear to be downstream late events in endotoxic shock, we hypothesized that a common signaling pathway might mediate these effects. METHODS: Using a guinea pig model of endotoxic shock, we investigated the effect of genistein and tyrphostin AG 556 on the cardiac action potential. Adult Hartley guinea pigs (300 to 450 gm) were randomized into 2 treatment parts. In the chronic treatment part, guinea pigs were randomized to receive daily subcutaneous injection of one of the five agents: saline, genistein, tyrphostin AG 556, daidzein, and vehicle for 10 days. In the acute treatment part, these agents were administered by intraperitoneal injection 1 hour before endotoxic shock. The animals were then anesthetized and mechanically ventilated, and underwent 6-hour endotoxic shock or sham experiment. RESULTS: In the chronic treatment part, the plasma nitrate concentration, myocardial guanosine 3',5'-cyclic monophosphate (cGMP) content, and APD at 90% repolarization (APD90) of papillary muscle showed no difference in the five groups before endotoxic shock. After 6-hour endotoxic shock, the elevation of plasma nitrate concentration and myocardial cGMP content was found significant in the control, the daidzein, and the vehicle groups, but was blunted in the genistein and the tyrphostin groups. The shortening of APD90 of papillary muscle was also significant in the control, the daidzein, and the vehicle groups, but blunted in the genistein and tyrphostin groups. There were similar findings in the acute treatment part, except the weaker effect of genistein and tyrphostin. CONCLUSIONS: Genistein and tyrphostin AG 556, either administered chronically or acutely, significantly attenuate the cardiac APD shortening in endotoxic shock, presumably through the decrease in the plasma nitrate and the cardiac cGMP production. It is suggested that tyrosine kinase signaling plays an important role in the modulation of APD in endotoxic shock. PMID- 12636203 TI - The relationship between morning serum cortisol and the short ACTH test in the evaluation of adrenal insufficiency. AB - BACKGROUND: Morning serum cortisol examinations and short corticotropin (ACTH) tests are popular screening tests for unstressed patients suspected of having adrenal insufficiency. However, the correlation between morning serum cortisol and the peak cortisol response in the short ACTH test after intravenous injection of ACTH has not been studied before. This retrospective study examined the relationship among the mean basal morning cortisol level, the single random morning cortisol level and the peak cortisol level in the short ACTH test in evaluation of adrenal insufficiency. METHODS: In this retrospective study, we examined the relationship among the mean basal morning cortisol level and the peak cortisol level stimulated by intravenous injection of 250 microg synthetic ACTH in 106 unstressed patients with proven or suspected hypothalamic-pituitary adrenal disease. Plasma cortisol levels were determined by radioimmunoassay. The correlation of the basal morning cortisol to the short ACTH test was determined by linear regression analysis. The ACTH test was analyzed using the receiver operating characteristic (ROC) curve method, and the cut-off points for various sensitivity and specificity were calculated. RESULTS: The mean basal is highly correlated to peak cortisol response in the ACTH test (r = 0.7724, p < 0.0001). Iatrogenic Cushing's syndrome, caused by ingestion of herbs with illegal steroid additives, was the most common cause (60%) of adrenal insufficiency in the 52 patients with subnormal result. A mean basal morning serum cortisol of > or = 300 nmol/L excluded the possibility of adrenal insufficiency, and a level of < 110 nmol/L suggested adrenal insufficiency. A mean basal cortisol level of > or = 234 nmol/L predicted a normal cortisol response in the ACTH test with optimal sensitivity (80.6%) and specificity (91.4%). CONCLUSIONS: The mean morning cortisol level is a cost-effective screening test in predicting the results of the ACTH test. A mean morning cortisol level > or = 300 nmol/L in suspicious cases of adrenal insufficiency which are not due to acute central etiologies excludes the possibility of subnormal cortisol response to ACTH; whereas a level < 110 nmol/L is likely to have subnormal cortisol response to ACTH. The short ACTH test or insulin tolerance test should be performed in patients with a morning cortisol level 300 nmol/L or with clinical symptoms highly suggestive of adrenal insufficiency. PMID- 12636204 TI - Non-steroidal anti-inflammatory drug and antacid co-prescription in Taiwan: analysis of national insurance claims. AB - BACKGROUND: Antacids were usually co-prescribed with non-steroidal anti inflammatory drugs (NSAIDs), although no broad evidences were available as to the effects of antacids in preventing NSAID-associated gastropathy. We performed a survey of national insurance claims for outpatient services in Taiwan to determine the extent of this co-therapy nationwide. METHODS: The National Health Insurance Research Database supplied the sampling datasets for analysis. They represented 0.2% of the entire claims for outpatient medical services in 1999. Co prescribing was assessed as NSAIDs and antacids on the same prescription. The selection and grouping of NSAIDs followed the guidelines of the Anatomical Therapeutic Chemical (ATC) Classification System recommended by the World Health Organization. Only the oral drugs prescribed on regular visits were taken into account. RESULTS: In totally 425,442 prescriptions with 1,825,604 items of drugs, non-aspirin NSAIDs were present in 108,818 (25.6%) prescriptions and antacids in 235,252 (55.3%) prescriptions respectively. Furthermore, antacids were present in 71.3% of prescriptions that contained NSAIDs and in 49.8% of prescriptions that did not contain NSAIDs (p < 0.001). Significant association of NSAIDs and antacids existed in different specialties of prescribing physicians, but the co prescription rate (antacids in NSAIDs prescriptions) varied from the highest of 92.8% in the neurosurgery to the lowest of 49.8% in the pediatrics. Significant association of NSAIDs and antacids also existed at different levels of health care facilities, where the co-prescription rates were 80.9% at medical centers, 83.5% at regional hospitals, 87.4% at local hospitals, and 66.6% at primary care units. The subgroup of oxicams was more frequently co-prescribed with antacids than other subgroups (odds ratio = 1.51, p = 0.001). CONCLUSIONS: Concomitant prescription of oral non-aspirin NSAIDs and antacids was indeed a popular practice in Taiwan. Beside their effects in alleviating the NSAID-associated dyspepsia, the role of antacids in preventing NSAID-associated peptic ulcers or in masking the warning symptoms of these ulcers demands further evaluation. PMID- 12636205 TI - Limited feasibility of routinely analyzing fetal cells from maternal blood by using magnetic activated cell sorting and polymerase chain reaction for prenatal diagnosis. AB - BACKGROUND: To assess the feasibility of analyzing fetal cells from maternal circulation by using magnetic activated cell sorting (MACS) and polymerase chain reaction (PCR) for prenatal diagnosis. METHODS: Thirty-one high-risk (either advanced maternal age or abnormal serum Down screening) pregnant women (14-22 weeks) were enrolled. Twenty ml of venous blood from each woman after amniocentesis were pretreated with density gradient centrifugation and sorted by MACS with monoclonal antibodies: anti-CD71 (n = 26) or anti-GPA (n = 5). Nested PCR with Y-specific probes--Y1.5-Y1.8 (n = 10) and Amelogenin (n = 21) were then applied to the sorted nucleated red blood cells (NRBCs) for fetal sex determination. These results were compared with cytogenetic data. To assess the sensitivity of PCR, different proportions of known male and female cultured amniocytes were mixed and amplified for gender identification. RESULTS: Karyotypes were normal in all fetuses (18 females and 13 males). The proportions of NRBCs (in total cells) sorted by MACS--anti-GPA or anti-CD71 were 50% (2000 +/ 1500) and 85% (350 +/- 280), respectively. Accuracies of sex determination by PCR-Amelogenin or Y1.5-Y1.8 were 76.2% (16/21) and 50% (5/10), respectively. Three cases resulted in PCR failure. Assay of nested PCR inferred that after cell sorting, existence of at least 20% of male fetal cells mixed in maternal blood circulation was required for prenatal diagnosis under current methodology. CONCLUSIONS: We confirmed the existence of fetal NRBCs in maternal blood during pregnancy. The low accuracy of sex determination (76.2%) may be attributed to contamination of either maternal NRBCs or non-NRBCs. No conclusive data, however, so far demonstrates the ideal marker to identify the origin of NRBCs. Without specific fetal cell marker and more sophisticated fetal cell analysis methodologies, in our experience, the feasibility of routinely analyzing fetal cells from maternal blood for prenatal diagnosis is limited. PMID- 12636206 TI - In vitro growth inhibition by indomethacin on human oral squamous cell carcinoma lines synergistically suppressed by all-trans retinoic acid correlating to apoptosis. AB - BACKGROUND: Indomethacin, an NSAID capable of inhibiting the effect of both cyclooxygenase and lipooxygenase, has been reported to repress the growth of breast cancer, skin cancer and head & neck cancer, etc. Inhibition in the some cell lines of oral squamous cell carcinoma (OSCC) has also been reported. The purpose of this study was primarily to explore the cellular response of human OSCC lines after indomethacin or retinoic acid (RA) treatment and its correlation to apoptosis phenomenon. METHODS: Five human OSCC cell lines--KB, SCC15, SCC25, OEC-M1 and OC2--were used for this in vitro study. By direct cell number counting, the cellular response was observed under incremental indomethacin concentrations of 50 microM, 100 microM, 200 microM and 400 microM, in order to select the most appropriate concentration for further study. Then 200 microM indomethacin and all-trans RA at 1 microM were used in the 2nd experiment to explore the intensity of their inhibitory effects individually and potential synergistic inhibition when exerted together. While in the 3rd part, TdT-mediated dUTP nick-end labeling (TUNEL) method was used for in situ apoptosis assay to see if the apoptosis rate varied with these two agents. RESULTS: All 5 cell lines constantly showed growth suppression with positive dosage effect of indomethacin. Synergistic inhibition by combined treatment of indomethacin and RA was seen in RA responsive lines of SCC15 and SCC25, whereas other RA-resistant clones showed no synergism of this combined treatment. The in situ detection of apoptosis by TUNEL assay revealed a significantly higher ratio of apoptotic cells in the indomethacin/RA treated SCC15 and SCC25 than in controls. CONCLUSIONS: The study provides the value of further exploration on the mechanism of how indomethacin inhibiting cancer cell growth and how RA-sensitive OSCC cell lines are synergistically suppressed by conjoint treatment of RA and indomethacin. This study also highlights the value to see how the apoptotic pathway responds differently to the indomethacin/RA treatment. PMID- 12636207 TI - Segmental pancreatectomy for benign tumor of the pancreas. AB - BACKGROUND: Enucleation of pancreatic tumor has the potential risk for damage of the main pancreatic duct. Benign tumors located in the neck or body of the pancreas are usually removed by left (spleno-) pancreatectomy or pancreatoduodenectomy. Standard pancreatic resection may result in serious loss of normal pancreatic parenchyme and impairment of pancreatic function. The aim of this study is to evaluate the results of segmental pancreatectomy, a limited resection of the midportion of the pancreas, in patients with benign tumor of the pancreas. METHODS: Four patients with benign tumor over pancreatic neck or body were treated with segmental pancreatectomy after pathological confirmation by frozen section. The proximal stump was closed and distal stump was anastomosed with a Roux-en-Y pancreaticojejunostomy. Clinical evaluation, routine blood sugar, stool fat examination and abdominal ultrasonography were performed for their follow up. RESULTS: Segmental pancreatectomy was satisfactory in these four patients. The pathologic examinations revealed serous cystadenoma. No mortality was noted. Minor pancreatic fistula was found in three of them and was treated conservatively. No obvious exocrine insufficiency was noticed. One patient had diabetes mellitus before operation, which was persisted postoperatively. CONCLUSIONS: Segmental pancreatectomy is a safe and effective alternative to major pancreatic resection in selected patients with benign tumor of the pancreas. This procedure has a surgical risk similar to that of the standard operation, but preserves more pancreatic tissues, which may prevent pancreatic function impairment. PMID- 12636208 TI - Hereditary hemorrhagic telangiectasia. AB - Hereditary hemorrhagic telangiectasia (HHT), also known as Osler-Weber-Rendu disease, is a rare disorder characterized by arteriovenous communications in visceral organs. The diagnosis of HHT consists of recurrent epistaxis, mucocutaneous telangiectasis, visceral vascular lesion and familial occurrence. HHT can be definitely diagnosed with the presence of all these three criteria. The prevalence of liver involvement of HHT was reported to range from 8 to 31%. Herein, we present a 75-year-old male who was diagnosed as having HHT with liver involvement, based on the findings of recurrent epistaxis, mucosal telangiectasis on the lower lip and hepatic arteriovenous malformation. The clinical presentations of this patient are discussed, and the literature is reviewed. PMID- 12636209 TI - Acute pancreatitis complicated with periappendicitis. AB - Colonic complications of severe acute pancreatitis are quite uncommon and always occur in the transverse colon and splenic flexure. Here we report the case of a 47-year-old male with mild acute pancreatitis (Ranson's score 1) who suffered from acute right lower quadrant pain during hospitalization. After conservative treatment failed, he underwent open appendectomy under the impression of acute appendicitis. However, the pathology revealed only periappendicitis. Small bowel ileus was noted on plain film of the abdomen and a high ascitic lipase level was found during operation suggesting that the periappendicitis resulted from the spreading of the pancreatic inflammatory exudate via the small bowel mesentery route. This report suggests that although rare, periappendiceal involvement mimicking acute appendicitis remains possible in even mild acute pancreatitis. PMID- 12636210 TI - Aggressive natural killer cell lymphoma/leukemia. AB - We report a case of aggressive natural killer (NK) cell lymphoma/leukemia in a 70 year-old woman presenting with fever, hepatosplenomegaly, retroperitoneal lymphadenopathy, and elevated serum CA19-9. The patient died 4 days after diagnosis. Neoplastic NK cells were identified in the blood and retroperitoneal lymph node biopsy. Their phenotypes were confirmed by extensive flow cytometric and immunohistochemical studies. In situ hybridization for Epstein-Barr virus (EBV)-associated RNA (EBER) was positive. Various forms of NK cell neoplasm were reviewed and discussed. PMID- 12636211 TI - Acute wound healing an overview. AB - The ability to heal an injury is a biologic necessity for all organisms, with mammals lagging in proficiency when compared with lower life forms that have the ability to regenerate differentiated structures. Technology and increased scientific knowledge have established a coordinated interplay that has improved the ability to manage wounds in a logical manner, and, on occasion, to accelerate the healing process. Insight into the complex chain of events leading to the formation of scar is a necessity for every individual who attempts wound management. PMID- 12636212 TI - Fetal wound healing current perspectives. AB - Early in gestation, fetal wounds are capable of healing scarlessly. Scarless healing in the fetus is characterized by regeneration of an organized dermis with normal appendages and by a relative lack of inflammation. Although there is a transition period between scarless and scar-forming repair, scarless healing also depends on wound size and the organ involved. The ability to heal scarlessly, furthermore, appears to be intrinsic to fetal skin. Unique characteristics of fetal fibroblasts, inflammatory cells, extra-cellular matrix, cytokine profile, and developmental gene regulation may be responsible for the scarless phenotype of early gestation fetal wounds. With the current knowledge, only minimal success has been achieved with the topical application of neutralizing antibodies, antisense oligonucleotides, and growth factors to improve wound-healing outcomes. Thus, further investigation into the mechanisms underlying scarless repair is crucial in order to devise more effective therapies for scar reduction and the treatment of cirrhosis, scleroderma, and other diseases of excessive fibrosis. PMID- 12636213 TI - Assessing and controlling wound infection. AB - Quantitatively, wounds harboring bacteria that exceed 105 colony-forming units per gram are considered infected wounds. There are acute wounds and chronic wounds, and the approach to controlling the infection is similar. Although the granulating bed may be avascular, systemic anti-infectives are employed adjunctively to circumvent systemic infection. The main armamentarium of the attack is the use of topical anti-infectives, which invade the bacteria where they reside, and, consequently, reduce their numbers and promote wound healing. PMID- 12636214 TI - Wound healing and diabetes mellitus. AB - Diabetes mellitus is one of the major contributors to chronic wound healing problems. When diabetic patients develop an ulcer, they become at high risk for major complications, including infection and amputation. The pathophysiologic relationship between diabetes and impaired healing is complex. Vascular, neuropathic, immune function, and biochemical abnormalities each contribute to the altered tissue repair. Despite treatment of these chronic wounds, which involves tight glucose control and meticulous wound care, the prognosis for their healing is quite poor. Newer modalities that deliver natural or engineered growth factors show a great deal of promise. All of the studies clearly show the continued need for well-controlled clinical trials. PMID- 12636215 TI - Impairments to wound healing. AB - Impaired wound healing is a complication faced by all physicians, regardless of their field of practice. Plastic surgeons are frequently called on to help treat patients who fail to heal properly. Therefore, plastic surgeons must be well versed in the intrinsic and extrinsic factors that can impair wound healing, such as nutrition, drugs, radiation, smoking, and hypoxia. Only by limiting detrimental factors can wound healing progress in a beneficial fashion. PMID- 12636216 TI - Cytokine manipulation of the wound. AB - Most individuals expect that healing is an inevitable outcome; wound healing is taken for granted. Although wound healing is perceived as inevitable, it can be fraught with problems and altered at many points. In the past, optimization of wound healing focused on minimizing contamination, accurate tissue approximation, and providing protection. With the advent of recombinant technology, optimization can now include manipulation of the molecular and cellular wound environment. Although the exact manipulative scheme has not yet evolved, it is clear from the multiple attempts reported in this article that understanding and progress is being made. PMID- 12636217 TI - Hyperbaric oxygen and wound healing. AB - Problem wounds, which fail to respond to traditional medical and surgical therapy, can be challenging to the plastic surgeon. Surgical, outpatient, and inpatient wound care costs can be exorbitant. Indirect costs, such as those related to patient productivity, disability, and premature death, can also be significant. The underlying problem in failure of a wound to heal is usually hypoxia and infection. HBO treatments in selected patients can facilitate healing by increasing tissue oxygen tension, thus providing the wound with a more favorable environment for repair. Therefore, HBO therapy can be an important component to any comprehensive wound care program. PMID- 12636218 TI - Fibroproliferative scars. AB - Fibroproliferative scars remain an ongoing clinical challenge. Both hypertrophic scars and keloids require multimodal therapy toachieve partally successful treatment. At the present time incomplete understanding about the pathogenesis of fibroproliferative scars makes targeted, mechanistic treatment impossible. As understanding of these abnormal wound problems increases, more effective treatments will likely be available. Until that time, clinicians must utilize existing knowledge to treat patients while continuing to experiment with new approaches. PMID- 12636219 TI - Tissue engineering. AB - Tissue engineering will potentially change the practice of plastic surgery more than any other clinical specialty. It is an interdisciplinary field that promises new methods of tissue repair. There has been more than $3.5 billion invested in this field since 1990. Relevant areas of progress include advanced computing, biomaterials, cell technology, growth factor fabrication and delivery, and gene manipulation. Beneficial clinical techniques will emerge from continued investigation in each of these areas. Techniques that are developed must be scaled up to industry with products cleared by regulatory agencies and acceptable to clinicians and patients. A goal of tissue engineering is to change clinical practice, yielding improved patient outcomes and lower costs of care. PMID- 12636220 TI - Visual adaptation. PMID- 12636221 TI - Perceived contrast following adaptation: the role of adapting stimulus visibility. AB - The issue of whether contrast adaptation can reduce the perceived contrast of gratings oriented orthogonal to the adapting stimulus to a greater extent than parallel gratings has been the subject of considerable debate (Snowden and Hammett, 1992; Ross and Speed, 1996). We compared the reductions in perceived contrast of various test gratings oriented parallel and orthogonal to the adapting stimulus across a range of spatial frequencies (2.25-9 c/deg) and adaptation contrasts (0.19-1.0). Our results show that when the adapting stimulus is low in contrast, parallel adaptation effects are always greater than the effects of orthogonal adaptation. When the adapting contrast is increased, however, the difference between parallel and orthogonal effects is reduced. Further increases in adapting contrast can produce a situation where cross orientation adaptation effects exceed iso-orientation effects. This was observed at low spatial frequencies (2.25 and 4.5 c/deg) only. The difference in the pattern of results obtained at low and high spatial frequencies can be explained in terms of the adapting stimulus visibility. We conclude that cross-orientation adaptation effects can be greater than iso-orientation effects, but only when the adapting stimulus is highly suprathreshold. PMID- 12636222 TI - Evidence for spatio-temporal selectivity in attentional modulation of the motion aftereffect. AB - An ignored region of the visual field might be monitored by an intermittent full visual analysis or by a more continuous but restricted analysis. We investigated which type of process is more likely in early vision by studying the effects of diverting attention on adaptation to a range of spatial (0.5, 2, 4. and 6 c/deg) and temporal (1.5 and 10 Hz) frequencies. During adaptation, subjects either fixated an unchanging digit (normal attention). or named the sequence of changing digits which formed the fixation point (diverted). The test field was always a static version of the adapting field, and the strength of adaptation was measured through the velocity and duration of subsequent Motion Aftereffects (MAEs). When attention during adaptation was normal MAE durations rose with spatial frequency for the 1.5 Hz stimuli, and declined with spatial frequency for the 10 Hz stimuli. When attention was diverted from the 10 Hz stimuli, MAE durations and velocities fell by a similar amount at all spatial frequencies. However, for the 1.5 Hz stimuli, the effects of diversion were very small at 0.5 c/deg, and rose progressively with spatial frequency, so that MAE reductions were largest at 6 c/deg. It appears that diversion hardly affects the encoding of coarse, slow stimuli, but attenuates the encoding of finer and/or faster stimuli. This is consistent with the idea that during diversion the visual system monitors the scene continuously, but over a restricted range of spatial and temporal scales. PMID- 12636223 TI - Increased visual after-effects in migraine following pattern adaptation extend to simultaneous tilt illusion. AB - Much previous research into visual processing in migraine supports a model of abnormal cortical processing, in between the headache attacks, that is characterised by hyperexcitability, heightened responsiveness, a lack of habituation and/or a lack of intra-cortical inhibition. Shepherd (2001) reported two adaptation studies that challenged this view, one using the tilt after effect, the second using the motion after-effect. Models of cortical function in migraine based on hyperexcitability and a lack of inhibition lead to specific predictions in an adaptation study: there should have been smaller after-effects in people with migraine than in people without. Both experiments, however, revealed larger after-effects in the migraine group than in the control group. Here, these results are extended to the simultaneous tilt illusion and an identical pattern of results was obtained: there were consistently larger effects in the migraine group than in the control group. The results from the three experiments are not consistent with a lack of inhibition in migraine. The results are discussed in terms of alternative models of cortical function, including a lack of excitation and reduced central energy reserves. PMID- 12636224 TI - Contrast adaptation may enhance contrast discrimination. AB - Whether contrast adaptation may enhance contrast discrimination is a question that has remained largely unresolved because of conflicting empirical evidence. Greenlee and Heitger (1988), for example, reported that contrast discrimination may be enhanced after contrast adaptation, while Maattanen and Koenderink (1991) did not. This paper aimed to account for the different conclusions reached by these independent researchers by manipulations of key differences that exist between the two studies. It is shown that contrast discrimination may be enhanced after adaptation, but that these effects can vary markedly across subjects and test conditions. Enhancements in contrast discrimination are reported to be significant when adapting and testing at low levels of contrast, but just significant at higher levels of contrast. For high contrast signals; enhancements are shown to be independent of temporal frequency but dependent upon viewing conditions. Under binocular viewing conditions, enhancements in contrast discrimination thresholds are shown to be significantly higher than under monocular viewing conditions. It is suggested that the different conclusions reached by Greenlee and Heitger and by Maattanen and Koenderink may be explained by their respective differences in viewing conditions. The former study used binocular, while the latter study used monocular viewing with an occluding eyepatch. PMID- 12636225 TI - Shading and texture: separate information channels with a common adaptation mechanism? AB - We outline a scheme for the way in which early vision may handle information about shading (luminance modulation, LM) and texture (contrast modulation, CM). Previous work on the detection of gratings has found no sub-threshold summation, and no cross-adaptation, between LM and CM patterns. This strongly implied separate channels for the detection of LM and CM structure. However, we now report experiments in which adapting to LM (or CM) gratings creates tilt aftereffects of similar magnitude on both LM and CM test gratings, and reduces the perceived strength (modulation depth) of LM and CM gratings to a similar extent. This transfer of aftereffects between LM and CM might suggest a second stage of processing at which LM and CM information is integrated. The nature of this integration, however, is unclear and several simple predictions are not fulfilled. Firstly, one might expect the integration stage to lose identity information about whether the pattern was LM or CM. We show instead that the identity of barely detectable LM and CM patterns is not lost. Secondly, when LM and CM gratings are combined in-phase or out-of-phase we find no evidence for cancellation, nor for 'phase-blindness'. These results suggest that information about LM and CM is not pooled or merged--shading is not confused with texture variation. We suggest that LM and CM signals are carried by separate channels, but they share a common adaptation mechanism that accounts for the almost complete transfer of perceptual aftereffects. PMID- 12636226 TI - A parametric account of contrast adaptation on contrast perception. AB - The effect of contrast adaptation on perceived contrast is assessed by contrast matching spatially adjacent sinusoidal gratings of similar spatial frequency, but different contrast and orientation. The main empirical question asked is why a high contrast orthogonal adaptor appears to amplify contrast signals through an increase in the slope of the contrast matching function but does not affect the threshold contrast at which a grating is detected. To explain this effect of adaptation, the Naka- Rushton receptor equation is employed as a description of the visual system's contrast response function. It is reported that the effects of adaptation may be described by three isotropic components, namely, signal amplification, division and addition, and one orientation specific component of subtraction. By collating the predictions made by the Naka-Rushton receptor equation with existing psychophysical data, it is shown that the magnitude of the isotropic components of adaptation increase with the contrast of the adapting signal. The orientation specific effect, however, is shown to saturate at relatively low adapting contrast levels. This saturation appears to be inconsistent with the commonly held view that the orientation specific effect represents a functional strategy used by the visual system to combat the problem of neural saturation in response firing rates. PMID- 12636227 TI - Autoimmune neutropenia. PMID- 12636228 TI - Telomere shortening, atherosclerosis, and metabolic syndrome. PMID- 12636229 TI - Therapy for lymphocytic adenohypophysitis. PMID- 12636230 TI - The influenza vaccination and neurological complications. PMID- 12636231 TI - Reactivation of immanent herpes group virus. PMID- 12636232 TI - Pleiotropic functions of erythropoietin. AB - Erythropoietin (EPO) produced by the fetal liver and adult kidney is an essential stimulator of erythropoiesis. EPO production is regulated through hypoxic activation of gene transcription and possibly hypoxia-induced stabilization of its mRNA. In the liver of early embryos in which EPO production poorly responds to hypoxia, retinoic acid may be an important stimulator. In this decade, new sites of EPO production have been found: central nervous system and reproductive organs. These tissues have a paracrine and/or autocrine system of EPO, which is independent of the endocrine system (kidney/bone marrow) in adult erythropoiesis. In the central nervous system, astrocytes are the main producers of EPO, while EPO receptor is expressed in neurons. EPO protects neurons from a various types of damage. The uterine EPO is likely involved in the estrogen-dependent angiogenesis of the endometrial layer. The possible functions of EPO in other tissues and tissue-characteristic regulation of EPO production are also discussed in this review. PMID- 12636233 TI - Telomere shortening of peripheral blood mononuclear cells in coronary disease patients with metabolic disorders. AB - OBJECTIVE: Telomere shortening is correlated with cell turnover and aging, but it has been recently suggested to occur not only by aging but by several biochemical factors of metabolic disorders predisposing to atherosclerosis. PATIENTS AND METHODS: We compared telomere length of peripheral blood mononuclear cells of patients with the metabolic disorders, hypercholesterolemia (HC) and diabetes mellitus (DM), according to the presence or absence of coronary diseases. RESULTS: The results demonstrated that HC and/or DM patients with coronary diseases have significantly shorter telomere length than healthy controls (p = 0.0014). CONCLUSION: Telomere shortening may be involved in the mechanisms that promote coronary diseases under some circumstances of metabolic disorders. PMID- 12636234 TI - Selenium deficiency in a patient with Crohn's disease receiving long-term total parenteral nutrition. AB - We report a case of selenium deficiency in a patient with Crohn's disease on long term total parenteral nutrition (TPN). She manifested lassitude of the legs, discoloration of the nail beds, and macrocytosis. Since her plasma selenium level was found to be below the measurable level, we diagnosed this case as selenium deficiency. After intravenous administration of sodium selenite, her symptoms were reversed. Careful attention should be paid to selenium deficiency when a patient receives long-term TPN; supplementary administration of selenium via TPN may be required because selenium is often not routinely added to TPN formulations. PMID- 12636236 TI - Treatment of lymphocytic hypophysitis by high-dose methylprednisolone pulse therapy. AB - A 26-year-old woman 3 months post-partum was admitted to our hospital suffering from gross visual disturbance. Magnetic resonance imaging (MRI) revealed a pituitary mass, extending into the suprasellar cistern, with intense gadolinium enhancement. Lymphocytic hypophysitis (LHy) was suspected, and the patient received high dose methylprednisolone pulse therapy (HDMPT). Her visual disturbance was dramatically ameliorated on the first day following initiation of HDMPT, and MRI revealed marked mass reduction. Her pituitary function recovered 6 months after therapy. This case report suggests that HDMPT proved effective for mass reduction of severe LHy and could obviate the need for a useless surgery. PMID- 12636235 TI - Usefulness of massive oral nicorandil in a patient with variant angina refractory to conventional treatment. AB - A 67-year-old man, who was previously diagnosed with vasospastic angina and treated with standard therapy, was admitted to our hospital because of recurrent chest pain refractory to sublingual nitroglycerin. Admission electrocardiography revealed ST segment elevation in II, III and aV(F), and his symptoms were relieved by intravenous bolus administration of nicorandil. He was diagnosed to have active variant angina, and remained symptomatic even after treatment with calcium antagonists and nitrates at optimal doses. Intravenous bolus administration of nicorandil was consistently effective to relieve his symptoms. Anginal attack was finally prevented by massive oral nicorandil in addition to conventional treatment. PMID- 12636237 TI - Type 2 diabetes complicated by multiple pyomyositis. AB - A 40-year-old man was hospitalized due to fever, muscular swelling and pain. He had poorly controlled diabetes with many dental caries and repeated periodontitis. CT revealed multiple intramuscular abscesses; administration of antibiotics and pus drainage were performed. Intraoral infection was suspected as the route of infection of pyomyositis, and a total of six teeth was extracted. In the clinical treatment of diabetic patients, it is important to instruct patients to routinely check for the presence of traumatic injuries of the lower extremities, and to have routine check-ups and dental care to check for dental caries or periodontitis. PMID- 12636238 TI - Granular cell tumor of the mediastinum. AB - Mediastinal granular cell tumor is an exceptionally rare tumor; there are only a few previously reported cases. We report a 21-year-old man with a granular cell tumor in the right posterior mediastinum. Chest computed tomography scans and magnetic resonance imaging revealed a 15x20 mm well circumscribed, homogeneous mass, which was closely associated with the vertebral column. The T1-weighted magnetic resonance image showed low intensities, and moderate intensities were found on the T2-weighted magnetic resonance image. Gd-DTPA enhancement was remarkable on the tumor surface. The present case differed from ordinary neurogenic tumors in magnetic resonance imaging findings. Thoracoscopic resection of the tumor was performed. The tumor was associated with the sympathetic nerve. Histologic analysis revealed a benign granular cell tumor. PMID- 12636239 TI - Multicentric Castleman's disease manifesting in the lung: clinical, radiographic, and pathologic findings and successful treatment with corticosteroid and cyclophosphamide. AB - Multicentric Castleman's disease (MCD) is an uncommon and often incurable lymphoproliferative disorder. There has been some recent evidence that rare cases of MCD manifest diffuse lung involvement, but the features in these cases are not well characterized. We report just such a biopsy-proven case of MCD with typical laboratory abnormalities including serum interleukin-6 elevation and characteristic high-resolution CT findings. Immunopathologically, the features of the lung tissue resembled those of lymphocytic interstitial pneumonia with predominant infiltration of B cells and plasma cells. In addition, the abnormal appearance of B cells in bronchoalveolar lavage fluid was of diagnostic value. Although MCD is often refractory to treatment including corticosteroid, chemo- and immuno-therapy, we show successful treatment with corticosteroid and cyclophosphamide and 4 years of complete remission. PMID- 12636240 TI - Secondary pulmonary alveolar proteinosis associated with myelodysplastic syndrome. AB - A 47-year-old man, who had been diagnosed as myelodysplastic syndrome (MDS), complained of a severe cough and a high-grade fever. Chest CT disclosed scattered small nodules and ground-glass opacities with interlobular septal thickening in both lung fields and a mass lesion in the right lower lobe. Pathological findings of the ground-glass opacities and the mass lesion obtained by video-assisted thoracoscopic surgery revealed the accumulation of eosinophilic amorphous material in the alveoli and confirmed the diagnosis of pulmonary alveolar proteinosis (PAP). Autoantibodies against granulocyte-macrophage colony stimulating factor (GM-CSF) in sera were below sensitivity, while the GM-CSF level was elevated in bronchoalveolar lavage fluid. He was diagnosed as secondary PAP associated with MDS. PMID- 12636241 TI - Neurologic complications associated with influenza vaccination: two adult cases. AB - We describe two adult cases of neurologic complications occurring after influenza vaccination. The first case was a 62-year-old man who experienced convulsions 5 days after vaccination, and the second case was a 70-year-old man who exhibited paraplegia 7 days after vaccination. Diagnoses of acute disseminated encephalomyelitis and transverse myelitis with acute motor axonal neuropathy were made, respectively, and steroid pulse therapy and intravenous gamma globulin therapy alleviated the patients' symptoms. Although the efficacy and cost benefit of influenza vaccination have been widely accepted, such neurologic complications might occur in the elderly or even in adults. PMID- 12636242 TI - A unique case of sporadic Creutzfeldt-Jacob disease presenting as progressive supranuclear palsy. AB - We report a Japanese case of sporadic Creutzfeldt-Jakob disease (CJD) presenting as progressive supranuclear palsy. For 2 years after onset, neurological deficits had slowly progressed but neither myoclonus nor periodic synchronous discharge was observed. Diffusion-weighted image (DWI) showed unique high signal lesions in the bilateral frontal cortex, left parietooccipital and occipital cortices, but there was nearly no change eight months later. Needle biopsy revealed deposition of prion protein of a patchy/perivacuolar type with spongiform degeneration. Thus, the phenotype of sporadic CJD seems variable and DWI should be performed, even in atypical cases lacking the characteristics of CJD. PMID- 12636243 TI - Probable sporadic Creutzfeldt-Jakob disease with valine homozygosity at codon 129 and bilateral middle cerebellar peduncle lesions. AB - We describe a 67-year-old Japanese man with probable sporadic Creutzfeldt-Jakob disease (CJD) who had valine homozygosity at codon 129, a rarity in the Japanese. T2-weighted magnetic resonance imaging (MRI) detected high-intensity lesions in the bilateral middle cerebellar peduncles and basal ganglia as well as cerebellar and cortical atrophy. He developed cerebellar ataxia and subsequent mental deterioration, myoclonus, and periodic synchronous discharge as shown in an electroencephalogram. Cerebrospinal fluid examination showed a high level of neuron-specific enolase and a positive immunoassay for the 14-3-3 protein. He died of pneumonia 10 months after the initial symptoms appeared. Whether or not the genetic polymorphism increased his susceptibility to sporadic CJD is not clear because valine homozygosity at codon 129 is less than 1% in the normal Japanese population. Although there is no convincing evidence in the present case, the MRI findings of cerebellar peduncle changes, which are rare in CJD, suggest a kind of degeneration, demyelination, or both. PMID- 12636244 TI - Salazosulfapyridine induced hypersensitivity syndrome associated with reactivation of humanherpes virus 6. AB - A 22-year-old woman with ulcerative colitis developed skin eruptions, liver dysfunction, and atypical lymphocytes in the peripheral blood two weeks after she started taking salazosulfapyridine (SASP). Skin eruptions and liver damage were severe. Drug-induced lymphocyte stimulation test (DLST) for SASP was positive. She was diagnosed as having SASP-induced hypersensitivity syndrome (HS). Corticosteroid therapy was needed to suppress these reactions. The transient elevation of HHV-6 IgG titer paralleled the symptoms, which indicated that these reactions were associated with the reactivation of HHV-6. We suggest that HHV-6 IgG titer is one of the modalities for the diagnosis and the prediction of the clinical course of HS. PMID- 12636245 TI - Multibacterial sepsis in an alcohol abuser with hepatic cirrhosis. AB - An alcohol abuser with hepatitis C developed multibacterial sepsis. His mean 100% alcohol intake reached 400 ml/day. In January 2001, he suddenly experienced fever (39 degrees C) with no other symptoms. One week later, he was admitted to our hospital and was subsequently diagnosed with sepsis associated with four species of bacteria (Streptococcus constellatus, Fusobacterium mortiferum, Bacteroides thetaiotaomicron, and non-spore-forming anaerobic gram-positive bacillus). A drip infusion of imipenem/cilastatin was administrated, resulting in a successful therapeutic outcome. No underlying disorder was found except for gastric ulcers and hepatic cirrhosis. Damaged gastric mucosa was assumed to be the possible cause and route for the bacterial invasion. PMID- 12636246 TI - A new therapeutic strategy for streptococcal toxic shock syndrome: a key target for cytokines. AB - We searched for innovative treatments specifically targeted to the cytokines of three critically ill patients with streptococcal toxic shock syndrome (STSS). Direct hemoperfusion with polymyxin B-immobilized fiber improved the hemodynamics in all three patients who had developed shock at presentation. Continuous hemodiafiltration coupled with plasma exchange for simultaneous removal of several mediators of the inflammatory cascade improved outcome in case 2 and lessened the aggressive clinical course and prolonged survival in case 3. The application of these technologies appears to open new and interesting therapeutic options and may lead to improved survival in STSS. PMID- 12636247 TI - Effect of environmental conditions on proteins secreted by enterohemorrhagic Escherichia coli O26:H11. AB - Infections due to Shiga toxin-producing Escherichia coli (STEC) are responsible for severe diarrheal diseases in humans, and these bacteria have recently emerged as a leading cause of renal failure and encephalitis in children and the aged. In this study, we examined the environment-dependent production of proteins secreted from a strain of STEC O26:H11 by trichloroacetic acid precipitation, SDS-PAGE, Western blotting and N-terminal amino acid sequence analysis. Growth of bacteria in essential minimum medium (M9) led to the detection of secreted proteins of 104, 80,40, 37 and 25 kDa (P104, P80, P40, P37 and P25, respectively). When grown in serum-free MEM, only P104, P40, P37 and P25 were observed in supernatant fluids. Growth of the bacteria in Luria-Bertani broth (LB) enhanced the expression of P104, but the productions of the other proteins were remarkably reduced. CO2 increased the secretion of P80 and P37, but reduced the production of P104. N-terminal amino acid sequencing revealed that P104 was EspP of STEC, which was homologous to EspC of enteropathogenic Escherichia coli (EPEC), and both proteins belong to a subclass of the IgA protease family. P80, which was identified as EspE of STEC, was homologous to Tir of EPEC. P40, P37 and P25 were found to be highly homologous to the similarly sized EspD, EspB and EspA proteins, previously detected in culture supernatants of EPEC. Those proteins are thought to be STEC virulence factors. Sera were obtained from two patients, one with colitis and another with hemolytic uremic syndrome (HUS), caused by STEC O157:H7, to study immune response to secreted proteins. Our results suggested that Tir caused immune response following STEC disease. PMID- 12636248 TI - Niche-specific association of Aeromonas Ribotypes from human and environmental origin. AB - A total of 88 Aeromonas isolates from distinct locations and sources (39 from extraintestinal infections, 31 from diarrhoeic, ten from non-diarrhoeic faeces, all human, and eight from fresh water) were subjected to phenospecies identification, serotyping, ribotyping and detection of some virulence markers. The strains belonged to four different phenospecies marked by 19 O serogroups and 38 ribotypes. No strong correlation between these parameters was found, and no group, as defined by the typing methods, could be characterized with a particular set of virulence markers. There was a clear association of ribotypes with the source of the strains. Cluster analysis allowed the identification of a complex of ribotypes belonging to distinct but related sources, including clinical and environmental isolates. These results suggest that ribotyping may be an epidemiological tool suitable for the study of Aeromonas infections. PMID- 12636249 TI - Transposon-induced norfloxacin-sensitive mutants of Bacteroides thetaiotaomicron. AB - To elucidate the mechanism of norfloxacin (a fluoroquinolone) resistance of Bacteroides thetaiotaomicron, a member of the B. fragilis group, we isolated transposon-induced mutants sensitive to this agent using Tn4351. Four norfloxacin sensitive mutants showed reduced levels of resistance, at least, to ethidium bromide. Cloning and sequencing of three chromosomal fragments adjacent to Tn4351 from the mutants revealed that two partial open reading frames (orfs) were disrupted by a transposon. Amino acid sequences of partial orf products had strong homologies to those of Escherichia coli RecB and B. ovatus transketolase. Two mutants carried a recB homolog inserted by Tn4351 together with R751 (cointegration) and by itself (simple transposition) at the amino- and carboxyl terminal portions, respectively. Since mutations in recB produce E. coli cells sensitive to DNA-damaging treatments by quinolones, it is concluded that decreases of the minimum inhibitory concentrations (MICs) of the agents for B. thetaiotaomicron resulted from disruption of the recB homolog. Another mutant carried a transketolase gene inserted by Tn4351. There is no reasonable explanation why disruption of the transketolase gene caused a decrease of the MIC of norfloxacin for this organism, although Streptococcus pneumoniae RecP related to DNA recombination was reported to be transketolase. PMID- 12636250 TI - The reduction of Fusobacterium nucleatum in mice is irrelevant to the nitric oxide induced by iNOS. AB - Previously we reported that mice infected recurrently with live Fusobacterim nucleatum (Fn) synthesize a significant amount of NO between 12 hr and 24 hr after the Fn injection. We now investigated whether the NO has the capability of killing Fn, a gram-negative rod periodontal pathogen. The mice were divided into three groups: treated with live bacteria (LB), treated with heat-killed bacteria (HKB) and untreated: normal (N). The Fn reduction, NO production and cell number after Fn injection were then compared in these mice. In the LB group, no Fn was detected at 6 hr, whereas it was still detected in the HKB and N groups at 24 hr as assessed by both colony counts and PCR assays. A significant amount of NO was synthesized in the LB group at 24 hr after the Fn injection. Fn is not killed by SNAP-generated NO in vitro. An increase in the total cell number was accompanied by an increase of the neutrophil numbers in the LB group. Intracellular O2(-) generation (including ONOO(-)) was visualized using dihydrorhodamine (DHR)-123. The peak of O2(-) generation by PEC was shown to be at 3 hr in all 3 groups. The number of O2(-) positive cells in the LB group at 3 hr was remarkably high, and most of them were likely to be neutrophils. The Fn reduction would be performed cooperatively via oxygen dependent and oxygen independent mechanisms. Thus reactive oxygen species (ROS) included in the oxygen dependent mechanism appear to be important for Fn reduction. However the significant amounts of NO derived from the iNOS synthesized in the LB group between 12 hr and 24 hr after injection of LFn were not involved in the Fn reduction. PMID- 12636251 TI - Lactoferrin feeding augments peritoneal macrophage activities in mice intraperitoneally injected with inactivated Candida albicans. AB - Oral administration of lactoferrin (LF), an innate-defense protein present in exocrine secretions such as milk and in neutrophils, is reported to improve host protection against infections with microorganisms including pathogenic fungi, possibly due to an immunomodulatory effect. This study aimed to evaluate the effect of bovine LF feeding on peritoneal macrophage activities in mice intraperitoneally injected with inactivated Candida albicans. Time course analysis during the 14 days following Candida-priming revealed that LF administration slightly increased the number of peritoneal exudate cells, and significantly enhanced the production of superoxide anion (O2(-)) and nitric oxide (NO) by peritoneal macrophages at day 7. LF administration facilitated NO production and Candida hyphal-growth inhibition by macrophages derived from Candida-primed mice but not non-primed mice, suggesting that the action of LF is dependent on the immune status of the host. LF administration altered the kinetics of cytokines in the peritoneal lavage fluid of Candida-primed mice. Enhancement of cytokine levels by LF was observed for IL-12 at day 5 and IFN gamma at day 9, but not for TNF-alpha or IL-10. In conclusion, LF feeding augmented the activities of macrophages in a manner dependent on Candida-priming and these effects may be related to enhanced cytokine levels. PMID- 12636252 TI - Isolation of Escherichia coli O157:H7 from dung beetles Catharsius molossus. AB - In an epidemiological survey, Escherichia coli O157:H7 was isolated from the intestine 4 of 113 dung beetle Catharsius molossus captured below ground at Tongshan County, Jiangsu Province of China. In parallel, 10 strains of E. coli O157:H7 were isolated from fecal samples of 383 diarrhea patients from the same region. Most importantly, using pulsed field gel electrophoresis (PFGE) of chromosomal DNA restriction fragments and PCR method, we found that the PFGE pattern and virulence genes of beetle isolates were identical to those of the human isolates, such as Shiga-toxins (stx) and enterohemorrhagic Escherichia coli hemolysin A (EHEC-hlyA). Therefore, dung beetle might acquire pathogenic E. coli O157:H7 through contact with feces of domestic animals. PMID- 12636253 TI - Serum antibodies of periodontitis patients compared to the lipopolysaccharides of Porphyromonas gingivalis and Fusobacterium nucleatum. AB - Serum antibody titers against the lipopolysaccharides (LPSs) of Porphyromonas gingivalis and Fusobacterium nucleatum were compared between 9 periodontitis patients and 24 healthy persons. The IgG titers against the LPSs of P. gingivalis ATCC 33277(T) and W50 were clearly higher in the patients than in the healthy persons. However, IgM titers against the LPSs of P. gingivalis strains were relatively low, and no significant difference was observed between the patients and healthy persons. On the other hand, IgG and IgM titers against the LPS of Fusobacterium nucleatum JCM 8532(T) in some patients were significantly higher than those in the healthy persons, although the difference in IgG titers was not large compared to that of the LPS of P. gingivalis. These results suggest that the antibody measurement of patients' sera against the LPS of periodontal bacteria can be applied for the diagnosis of periodontitis. PMID- 12636254 TI - Antimicrobial susceptibilities of Shiga toxin-producing Escherichia coli isolates from pigs with edema disease in Japan. AB - Fifty-seven Shiga toxin-producing Escherichia coli (STEC) strains isolated from pigs with edema disease (ED) from 1997 to 2001 in Japan were examined for antimicrobial susceptibilities. The susceptibilities were compared with those of E. coli ATCC 23546 isolated from pig with ED in the 1950's. Consequently, the isolated STECs showed high susceptibility to peptides and bicozamycin in a way similar to the reference strain. On the other hand, the STECs showed low susceptibility to beta-lactams, tetracyclines, novobiocin, fosfomycin, trimethoprim, and old quinolones. It became clear that the susceptibilities of the isolated STECs had diminished in regard to antimicrobials. PMID- 12636256 TI - Selection of human immunodeficiency virus type 1 variants with an insertion mutation in the p6(gag) and p6(pol) genes under highly active antiretroviral therapy. AB - We detected several types of human immunodeficiency virus type 1 (HIV-1) variants with an insertion mutation in the p6(gag)and p6(pol) genes in eight of twenty-two (36.4%) patients who possessed drug-resistant viruses under highly active antiretroviral therapy (HAART). It was characteristic that a conserved proline rich motif "PTAPP" in the N-terminus of p6(gag) protein was completely or partially duplicated in all cases. Five among the eight cases were retrospectively investigated in terms of the occurrence of dynamic change in the gag gene between the inserted and wild-type HIV-1 in the course of HAART. The longitudinal analysis revealed the following: 1) The inserted-type viruses were selected over the wild-type during HAART in three cases in which the both types coexisted in the beginning of the therapy. 2) In two cases in which the inserted type HIV-1 alone was detected before the beginning of HAART, the inserted-type HIV-1 alone was continuously detected during the therapy. The inserted-type HIV-1 was also detected in four of thirty-nine (10.3%) therapy-naive patients. However, the frequency of inserted-type HIV-1 detection in the HAART-receiving patients is significantly higher than that in the therapy-naive patients (P = 0.02). These results suggest that this type of insertion mutation is a polymorphism of the p6(gag) and p6(pol) genes, however, it consequently gave an advantage on proliferation and/or survival of the HIV-1 variant under the presence of antiretroviral drugs. PMID- 12636255 TI - Tyrosine phosphorylation of a novel 100-kDa protein coupled to CD28 in resting human T cells is enhanced by a signal through TCR/CD3 complex. AB - For T cell activation, two signals are required, i.e., a T cell receptor (TCR)/CD3-mediated main signal and a CD28-mediated costimulatory signal. CD28 binds to its ligand (CD80 or CD86) and transduces the most important costimulatory signal. The cytoplasmic domain of the CD28 molecule, composed of 41 amino acids, does not contain any intrinsic enzyme activity. The cytoplasmic domain of CD28 is remarkably conserved among species and is associated with a number of signaling molecules that affect the main signal. We report here that a tyrosine phosphorylated 100-kDa protein (ppl00) was coupled to the CD28 cytoplasmic domain in Jurkat and human peripheral T cells. The pp100 was distinguished from other CD28 associated molecules such as Vav, STAT5, PI 3 kinase, Valosin-containing protein (VCP), Nucleolin, Gab2 (Grb2-associated binding protein 2), and STAT6. The tyrosine phosphorylation of pp100 coprecipitated with CD28 was enhanced by CD3 stimulation by the specific antibody, tyrosine phosphatase inhibitor and PKC activator. Tyrosine phosphorylation of pp100 was attenuated by the prior addition of PKC inhibitor. These findings indicate that pp100 is a novel tyrosine phosphorylated protein coupled to CD28 under continuous control of tyrosine phosphatases and might play a role in T cell activation augmented by a TCR/CD3-mediated main signal. PMID- 12636257 TI - Purification, cloning and characterization of variant LukE-LukD with strong leukocidal activity of staphylococcal bi-component leukotoxin family. AB - Staphylococcus aureus produces bi-component leukotoxins composed of non associated soluble proteins, S and F. Neither S nor F component alone is cytotoxic, but components together are active. These include Panton-Valentine leukocidin (PVL), gamma-hemolysin, LukE-LukD and others. Purification of leukotoxin from Staphylococcus aureus V8 strain (ATCC 27733) which does not have PVL genes, identified an F component with 100% identical to that of PVL in the first twenty-five N-terminal amino acids. Molecular cloning of this toxin obtained 2,595 nucleotides sequences containing two novel open reading frames for S and F. Deduced amino acid sequences of the S and F were respectively 91 and 94% identical to those of LukE and LukD. These were named variant of LukE-LukD (LukEv LukDv). The activity of the recombinant LukEv-LukDv to rabbit leukocytes was similar to that of recombinant PVL. LukEv-LukDv was hemolytic to rabbit red blood cells although the activity was only 8% of gamma-hemolysin, but PVL was not. These activities were quite different from the LukE-LukD which was reported no hemolytic and poorly cytotoxic to leukocytes compared to PVL. The lukEv-IukDv was found in 87% of clinical isolates of Staphylococcus aureus but lukE-lukD was not detected. These data demonstrate the existence of variant LukE-LukD in V8 strain (ATCC 27733). PMID- 12636259 TI - Individuals exhibiting conspicuous nevi (lentigo simplex) are resistant to allergic rhinitis/conjunctivitis (pollinosis), but those who do not show increased susceptibility to pollinosis. AB - We investigated 115 individuals in Tokyo from two groups of persons with conspicuous nevi on the head, neck, and arms, etc. (nevus-positive group) and persons who do not exhibit such nevi (nevi-negative group) interviewing them to determine whether they suffered from allergic rhinitis/conjunctivitis (pollinosis) and examining them for the presence and extent of nevi. There was a distinct correlation: only 5 persons among 85 individuals of the nevus-positive group suffered from pollinosis, and all 30 people in the nevus-negative group had pollinosis. Thus, individuals exhibiting conspicuous nevi are resistant to pollinosis, but those who do not show increased susceptibility to pollinosis. PMID- 12636258 TI - Epstein-Barr virus (EBV) nuclear antigen leader protein (EBNA-LP) forms complexes with a cellular anti-apoptosis protein Bcl-2 or its EBV counterpart BHRF1 through HS1-associated protein X-1. AB - Epstein-Barr virus (EBV) nuclear antigen leader protein (EBNA-LP) plays a critical role in EBV-induced transformation. An earlier report (Y. Kawaguchi et al., J. Virol. 74: 10104-10111, 2000) showed that EBNA-LP interacts with a cellular protein HS1-associated protein X-1 (HAX-1). The predicted amino acid sequence of HAX-1 exhibits similarity to that of another cellular protein Nip3 which has been shown to interact with cellular and viral anti-apoptotic proteins such as Bcl-2 and BHRF1, an EBV homolog of Bcl-2. Here we investigated whether HAX-1, like Nip3, interacts with Bcl-2 proteins and report the following. (i) A purified chimeric protein consisting of gluthathione S-transferase (GST) fused to BHRF1 (GST-BHRF1) or Bcl-2 (GST-Bcl-2) specifically pulled down HAX-1 transiently expressed in COS-7 cells. (ii) GST-BHRF1 or GST-Bcl-2 was not able to pull down EBNA-LP transiently expressed in COS-7 cells, whereas each of the GST fusion proteins formed complexes with EBNA-LP in the presence of RAX-1. These results indicated that EBNA-LP interacts with the viral and cellular Bcl-2 proteins through HAX-1, suggesting that EBNA-LP possesses a potential function in the regulation of apoptosis in EBV-infected cells. PMID- 12636260 TI - High molecular weight factor in FCS inhibits Helicobacter pylori VacA-binding to its receptor, RPTPbeta, on AZ-521. AB - VacA, a secretory product of Helicobacter pylori, binds to its cell surface receptor, receptor tyrosine phosphatase (RPTP) beta, leading to cytoplasmic vacuolization of gastric epithelial AZ-521 cells. VacA binding to the cell surface and VacA-dependent vacuolization were inhibited by cell culture media containing fetal calf serum (FCS). The high molecular weight fraction of FCS isolated by Superose 12 gel filtration chromatography inhibited VacA binding, whereas only weak effects were observed with other fractions. These data show that the high molecular weight fraction of FCS inhibits VacA action though its ability to block toxin binding to its receptor, RPTPbeta, on AZ-521 cells. PMID- 12636262 TI - Rate of evolution of the specific surface area of surface snow layers. AB - The snowpack can impact atmospheric chemistry by exchanging adsorbed or dissolved gases with the atmosphere. Modeling this impact requires the knowledge of the specific surface area (SSA) of snow and its variations with time. We have therefore measured the evolution of the SSA of eight recent surface snow layers in the Arctic and the French Alps, using CH4 adsorption at liquid nitrogen temperature (77 K). The SSA of fresh snow layers was found to decrease with time, from initial values in the range 613-1540 cm2/g to values as low as 257 cm2/g after 6 days. This is explained by snow metamorphism, which causes modifications in crystal shapes, here essentially crystal rounding and the disappearance of microstructures. A parametrization of the rate of SSA decrease is proposed. We fit the SSA decrease to an exponential law and find that the time constant alpha(exp) (day(-1)) depends on temperature according to alpha(exp) = 76.6 exp ( 1708/7), with Tin kelvin. Our parametrization predicts that the SSA of a snow layer evolving at -40 degrees C will decrease by a factor of 2 after 14 days, while a similar decrease at -1 degrees C will only require 5 days. Wind was found to increase the rate of SSA decrease, but insufficient data did not allow a parametrization of this effect. PMID- 12636261 TI - Efficient adeno-associated virus-mediated gene expression in human placenta derived mesenchymal cells. AB - Mesenchymal cells from various sources are pluripotent and are attractive sources for cell transplantation. In this study, we analyzed recombinant adeno-associated virus (rAAV)-mediated gene expression in human placenta-derived mesenchymal cells (hPDMCs), which reside in placental villi. After transduction of AV-CAG-EGFP, a rAAV expressing enhanced green fluorescence protein (EGFP), hPDMCs showed much higher level of EGFP expression than human umbilical vein endothelial cells or rat aortic smooth muscle cells. The number of EGFP-positive hPDMCs infected by AV CAG-EGFP alone did not increase significantly by coinfection of adenovirus, which enhanced expression level of the rAAV vector. Moreover, flow cytometric analysis showed discrete positive fraction of EGFP-expressing hPDMCs, which is about 15 20% of the cells infected with AV-CAG-EGFP. Therefore, some cell population in hPDMCs might be highly susceptible to rAAV-mediated gene transduction. In addition, stable EGFP expressions were observed in about 1% of hPDMCs infected with AV-CAG-EGFP at 4 weeks post-infection. Collectively, hPDMCs have characters favorable for rAAV-mediated gene expression. PMID- 12636263 TI - Global distribution and budget of PCBs and HCB in background surface soils: implications for sources and environmental processes. AB - This paper presents data from a survey of polychlorinated biphenyls (PCBs) and hexachlorobenzene (HCB) concentrations in 191 global background surface (0-5 cm) soils. Differences of up to 4 orders of magnitude were found between sites for PCBs. The lowest and highest PCB concentrations (26 and 97,000 pg/g dw) were found in samples from Greenland and mainland Europe (France, Germany, Poland), respectively. Background soil PCB concentrations were strongly influenced by proximity to source region and soil organic matter (SOM) content Most (>80%) of the estimated soil PCB burden remains in the "global source region" of the Northern Hemisphere (NH) temperate latitudes (30-60 degrees N) or in the OM-rich soils just north of that %SOM correlated with PCB and HCB in the global data set, with the correlation coefficients being greater for HCB and the lighter PCBs than for heavier homologues. OM-rich soils in the NH consistently contained the highest burdens; such soils are a key global compartment for these compounds. Evidence for global fractionation of PCBs was found in the subset of soils from latitudes north of the global source region but was not discerned with the global data set The full data set was used to estimate the burden for individual congeners/homologues in surface background soils and a global soil total PCB burden of 21,000 t. The significance of the inventory is briefly discussed in relation to the latest estimates of global production and atmospheric emission. PMID- 12636264 TI - Tiered approach for identification of a human fecal pollution source at a recreational beach: case study at Avalon Bay, Catalina Island, California. AB - Recreational marine beaches in California are posted as unfit for swimming when the concentration of fecal indicator bacteria (FIB) exceeds any of seven concentration standards. Finding and mitigating sources of shoreline FIB is complicated by the many potential human and nonhuman sources of these organisms and the complex fate and transport processes that control their concentrations. In this study, a three-tiered approach is used to identify human and nonhuman sources of FIB in Avalon Bay, a popular resort community on Catalina Island in southern California. The first and second tiers utilize standard FIB tests to spatially isolate the FIB signal, to characterize the variability of FIB over a range of temporal scales, and to measure FIB concentrations in potential sources of these organisms. In the third tier, water samples from FIB "hot spots" and sources are tested for human-specific bacteria Bacteroides/Prevotella and enterovirus to determine whether the FIB are from human sewage or from nonhuman sources such as bird feces. FIB in Avalon Bay appear to be from multiple, primarily land-based, sources including bird droppings, contaminated subsurface water, leaking drains, and runoff from street wash-down actvities. Multiple shoreline samples and two subsurface water samples tested positive for human specific bacteria and enterovirus, suggesting that at least a portion of the FIB contamination is from human sewage. PMID- 12636265 TI - Locating and quantifying PCB sources in Chicago: receptor modeling and field sampling. AB - Potential source contribution function (PSCF) modeling using polychlorinated biphenyl (PCB) concentrations measured in the Chicago area resolved three PCB source sectors. They were (i) the direction northwest of Chicago, (ii) the direction southwest of Chicago, and (iii) the south side of Chicago in the neighborhood of Lake Calumet. The area south of Chicago was further examined by taking upwind/ downwind samples near a landfill and sludge drying beds. Results identified the sludge drying beds and a large landfill as PCB sources to the atmosphere. Another PCB source identified in Chicago was a transformer storage yard. This site had the highest upwind/downwind concentration increments in this study (downwind PCB concentrations were more than 5 times those in the upwind air). These PCB sources were characterized in terms of inventories, emission rates, contributions, and PCB congener profiles (fingerprints). Preliminarily results indicate that the sludge may emit up to 90 kg/yr of PCBs to the air. This amount is probably not a significant contribution of PCBs to the Chicago atmosphere on the basis of dispersion modeling results and a simple box model. PMID- 12636266 TI - Caffeine, an anthropogenic marker for wastewater comtamination of surface waters. AB - The suitability of caffeine as a chemical marker for surface water pollution by domestic wastewaters was assessed in this study. Caffeine concentrations in influents and effluents of Swiss wastewater treatment plants (WWTPs, 7-73 and 0.03-9.5 microg/L, respectively) indicated an efficient elimination of 81-99.9%. Corresponding loads in untreated wastewater showed small variations when normalized forthe population discharging to the WWTPs (15.8 +/- 3.8 mg person(-1) d(-1)), reflecting a rather constant consumption. WWTP effluent loads were considerably lower (0.06 +/- 0.03 mg person(-1) d(-1)), apart from installations with low sludge age (< or = 5 d, loads up to 4.4 mg person(-1) d(-1)). Despite the efficient removal in most WWTPs, caffeine was ubiquitously found in Swiss lakes and rivers (6-250 ng/ L), except for remote mountain lakes (<2 ng/L; analytical procedure for wastewater and natural waters: SPE, GC-MS-SIM or GC-MS MS-MRM, internal standard 13C3-labeled caffeine). Caffeine concentrations in lakes correlated with the anthropogenic burden by domestic wastewaters, demonstrating the suitability of caffeine as a marker. A mass balance for Greifensee revealed that approximately 1-4% of the wastewaters had been discharged without treatment, presumably on rainy days when the capacity of WWTPs had been exceeded. For Zurichsee, it could be shown that the monthly inputs of caffeine correlated with precipitation data. The depth- and seasonal-dependent concentrations in this lake were adequately rationalized by a numerical model considering flushing, biodegradation, and indirect photodegradation via HO. radicals as elimination processes and caffeine inputs as fitting variables. PMID- 12636267 TI - Cd bioaccumulation by a freshwater bacterium, Rhodospirillum rubrum. AB - Cd bioaccumulation by Rhodospirillum rubrum, a Gram-negative freshwater bacterium, was studied in a synthetic medium. The free ion (Cd2+) was the best predictor of the Cd internalization fluxes. Representation of the short-term uptake fluxes as a function of [Cd2+] in the medium demonstrated a linear relationship, as would be expected for a rate-limiting, first-order internalization with a single transporter. Nonetheless, several different accumulation profiles were observed, depending on the Cd concentration. Cd uptake was regulated differently for concentrations above and below 10(-6) M (or was regulated only above [Cd2+] = 10(-6) M). Short-and long-term studies revealed that regulation was rapidly initiated for the highest Cd concentrations examined, effectively decreasing both adsorbed and internalized Cd. Anodic stripping voltammetry demonstrated that a Cd complexing ligand was produced within minutes upon exposure to 5 x 10(-6) M Cd2+ and that an extracellular sequestration of Cd was one mechanism regulating Cd uptake. Competition studies with other cations revealed a competitive inhibition of Cd uptake by Zn and an uptake enhancement in the presence of Mn and Cu. PMID- 12636268 TI - Degradation of racemic and enantiopure metalaxyl in tropical and temperate soils. AB - The degradation of the racemic mixture and the enantiomers of metalaxyl in typical soils from Germany and Cameroon has been studied. Formulated and unformulated R-metalaxyl were studied as well as racemic (rac) metalaxyl in controlled incubation experiments. The kinetics of the degradation or transformation were determined by means of reversed phase HPLC, while the enantiomeric ratios were measured by HPLC with a chiral Whelk O1 column. The degradation followed first-order kinetics (R2 > or = 0.96). Higher metalaxyl acid metabolite concentrations were found in German soil than in Cameroonian soil. The enantiomers of the fungicide each had different degradation rates in both soils, with half-lives ranging from 17 to 38 days. All forms of metalaxyl had lower degradation rates in the Cameroonian soil than in the German soil. The degradation. of the R-enantiomer was much faster than the S-enantiomer in the German soil and slower than the S-enantiomer in the Cameroonian soil suggesting that different microbial populations, which may be using different enzymes, have different degradation preferences. The results for the major differences in the degradation of the enantiomers may have some implication for the frequency of use as well as the environmental assessment for chiral pesticides. PMID- 12636269 TI - Reductive dehalogenation of halomethanes in iron- and sulfate-reducing sediments. 1. Reactivity pattern analysis. AB - The incorporation of reductive transformations into environmental fate models requires the characterization of natural reductants in sediments and aquifer materials. For this purpose, reactivity patterns (range and relative order of reactivity) for a series of 14 halogenated methanes were measured in iron- and sulfate-reducing sediments and two representative model systems: adsorbed Fe(II)/goethite [Fe(II)ads/alpha-FeOOH] and iron sulfide (FeS). Both Fe(II)ads and FeS are naturally occurring reductants. The strong similarity in reactivity patterns between the iron- and sulfate-reducing sediments suggests that the two share a common reductant despite their different chemical compositions (i.e., the sulfate-reducing sediment contained FeS). An orthogonal regression analysis of the halomethane transformation rate data in the sediment and model systems supports the assumption that a common mechanism for halomethane transformation exists between the sediments and the Fe(II)ads/alpha-FeOOH system and further corroborates the conclusion that Fe(II) adsorbed to Fe(III)-containing minerals is the dominant reductant in both sediment systems. Weak (0.5 N) and strong (6.0 N) acid extraction of the sediments indicated that solid-phase Fe(II) was 67% higher in the sulfate-reducing sediment than in the iron-reducing sediment, which is consistent with the observations that the halomethanes were transformed a factor of 3 times faster in the sulfate-reducing sediment and that Fe(II) was the dominant reductant. PMID- 12636270 TI - Reduction of uranium(VI) by mixed iron(II)/iron(III) hydroxide (green rust): formation of UO2 nanoparticles. AB - Green rusts, which are mixed ferrous/ferric hydroxides, are found in many suboxic environments and are believed to play a central role in the biogeochemistry of Fe. Analysis by U LIII-edge X-ray absorption near edge spectroscopy of aqueous green rust suspensions spiked with uranyl (U(VI)) showed that U(VI) was readily reduced to U(IV) by green rust The extended X-ray absorption fine structure (EXAFS) date for uranium reduced by green rust indicate the formation of a UO2 phase. A theoretical model based on the crystal structure of UO2 was generated by using FEFF7 and fitted to the data for the UO2 standard and the uranium in the green rust samples. The model fits indicate that the number of nearest-neighbor uranium atoms decreases from 12 for the UO2 structure to 5.4 forthe uranium-green rust sample. With an assumed four near-neighbor uranium atoms per uranium atom on the surface of UO2, the best-fit value for the average number of uranium atoms indicates UO2 particles with an average diameter of 1.7 +/- 0.6 nm. The formation of nanometer-scale particles of UO2, suggested by the modeling of the EXAFS data, was confirmed by high-resolution transmission electron microscopy, which showed discrete particles (approximately 2-9 nm in diameter) of crystalline UO2. Our results clearly indicate that U(VI) (as soluble uranyl ion) is readily reduced by green rust to U(IV) in the form of relatively insoluble UO2 nanoparticles, suggesting that the presence of green rusts in the subsurface may have significant effects on the mobility of uranium, particularly under iron-reducing conditions. PMID- 12636271 TI - Impact of metal sorption and internalization on nitrification inhibition. AB - The goal of this study was to explore the relationship between metal extracellular sorption, intracellular accumulation, and nitrification inhibition. Metal sorption on nitrifying biomass was rapid and could be described by linear partitioning with partition coefficients (Kp) of 20.3 +/- 0.1, 0.4 +/- 0.0, 0.1 +/- 0.0, and 0.2 +/- 0.0 L/g biomass chemical oxygen demand for Cu, Zn, Ni, and Cd, respectively. On the other hand, intracellular Zn, Ni, and Cd concentrations continued to increase with time beyond 12 h after metal addition, whereas intracellular Cu attained equilibrium after 4 h. Metal internalization kinetics could be described by an intraparticle diffusion model, with characteristic diffusion time constants (td) of 9.4, 64.6, 80.5, and 66.1 h for Cu, Zn, Ni, and Cd, respectively. Ultimate internalized percentages of the total cell-associated metal were 1.4 +/- 0.0, 4.3 +/- 0.5,7.6 +/- 1.0, and 2.7 +/- 0.2% for Cu, Zn, Ni, and Cd, respectively. Nitrification inhibition was not a function of the sorbed metal fraction but correlated well with intracellular Zn, Ni, or Cd fractions. An intraparticle diffusion model coupled with a saturation-type biological toxicity model fit the inhibition data for varying initial Cd concentrations and exposure periods. In contrast, no relationship between intracellular or sorbed Cu concentrations and nitrification inhibition was observed. In the presence of 1 mM Cu, less than 13.3 +/- 10.5% cells remained viable as compared to 72.8 +/- 7.5,104.8 +/- 1.7, and 84.7 +/- 7.0% (assumed 100% viable cells in metal-free control) in the presence of 1 mM Zn, Ni, and Cd, respectively. Hence, the observations that inhibition by metals such as Zn, Ni, and Cd is related to their intracellular fraction and the slow kinetics of metal internalization indicate that metal inhibition can easily be underpredicted from short-term batch assays. Furthermore, the inhibitory mechanism of Cu was very different from Zn, Ni, and Cd and may involve rapid loss of membrane integrity. PMID- 12636272 TI - Sediment-water distribution of organic contaminants in aquatic ecosystems: the role of organic carbon mineralization. AB - The distribution between sediments and water plays a key role in the food-chain transfer of hydrophobic organic chemicals. Current models and assessment methods of sediment-water distribution predominantly rely on chemical equilibrium partitioning despite several observations reporting an "enrichment" of chemical concentrations in suspended sediments. In this study we propose and derive a fugacity based model of chemical magnification due to organic carbon decomposition throughout the process of sediment diagenesis. We compare the behavior of the model to observations of bottom sediment-water, suspended sediments-water, and plankton-water distribution coefficients of a range of hydrophobic organic chemicals in five Great Lakes. We observe that (i) sediment water distribution coefficients of organic chemicals between bottom sediments and water and between suspended sediments and water are considerably greaterthan expected from chemical partitioning and that the degree sediment-water disequilibrium appears to follow a relationship with the depth of the lake; (ii) concentrations increase from plankton to suspended sediments to bottom sediments and follow an inverse ratherthan a proportional relationship with the organic carbon content and (iii) the degree of disequilibrium between bottom sediment and water, suspended sediments and water, and plankton and water increases when the octanol-water partition coefficient K(ow) drops. We demonstrate that these observations can be explained by a proposed organic carbon mineralization model. Our findings imply that sediment-water distribution is not solely a chemical partitioning process but is to a large degree controlled by lake specific organic carbon mineralization processes. PMID- 12636273 TI - Partition of fish pheromones between water and aggregates of humic acids. Consequences for sexual signaling. AB - High-molecular-weight suspended organic matter of soil and aquatic origins competes with water for the dissolution of relatively water insoluble organic substances. The same happens with microalgae and other organisms present in natural waters. Several pheromones, which play a specific role in the reproductive cycle of fish, are secreted to the water and are generally, if not always, molecules with hydrophobic or amphiphilic characteristics. The natural tendency of these pheromones to dissolve in suspended or deposited organic matter may cause their signaling function to be adversely affected. In this work we study the partition constants between water and organic reservoirs in suspension, Kh, of two fish pheromones, 4-pregnene-17 alpha, 20beta-diol-3-one (17,20beta-P) and prostaglandin F2alpha (PGF2alpha), and also of 4-pregnene-11beta,21-diol-3,20 dione 21-sulfate (21-P-sulfate), used as a proxy for sulfated steroid pheromones. Two types of organic reservoirs are employed: aggregates of suspended humic substances and negatively charged phospholipid vesicles. We find that the three compounds have high affinities for both types of aggregates. However, 17,20beta P, with pKh = 4.4 +/- 0.2, is the only one for which we may predict a significant decrease in availability in consideration of the normal content of dissolved organic matter in natural waters. Also to be considered is the fact that a relatively large amount of pheromones is retained and may be released at an inopportune moment. How significant these phenomena are in nature is not as yet clear, considering the variety of habitats in which fish spawn and the little that is known about the nature and mode of action of pheromones. PMID- 12636274 TI - Quantification of the interaction of Tc with dissolved boom clay humic substances. AB - Technetium-99 (Tc), a fission product of uranium-238, is an important radionuclide because of its long half-life and its high yield in radioactive waste. To elucidate the Tc geochemical behavior in reducing environments relevant to geological disposal and in the presence of humic substances (HS), experiments were set up that resulted forthe first time in the determination of an interaction constant for Tc with dissolved humic substances. A number of lab scale Boom Clay (a possible geological underground High-Level Radioactive Waste storage site in Mol, Belgium) batch experiments were set up, combining both different initial Tc(VII) concentrations and different solid/liquid ratios. On these batches several sequential extraction steps with HS-free synthetic Boom Clay water were performed. Equilibration times were fixed at 1 week for each extraction step. Tc(VII) was found to be readily reduced to Tc(IV) by the solid Boom Clay phase. This solid phase was able to sorb Tc(IV) to a very large extent (log Kd approximately 2.5-4.0), and two sorption sinks (one of which is humic substances) were detected. In solution, Tc(IV) was mainly associated with HS. Concentrations in solution were found up to the order of 2 x 10(-6) M. The results were quantitatively described as a competition for Tc(IV) between the solid phase and the dissolved HS (Schubert-like approach). It was concluded that a hydrophobic sorption of uncharged Tc(IV) species in solution would act as the dominating interaction mechanism with HS, with an interaction constant log K(HS) = 5.3 +/- 0.3. PMID- 12636275 TI - XAS speciation of arsenic in a hyper-accumulating fern. AB - The coordination environment and the redox speciation of arsenic in a newly discovered arsenic hyper-accumulating fern (Pteris vittata L) were investigated by X-ray absorption spectroscopy. This method allowed us to probe arsenic directly, i.e., with minimal sample preparation. The results indicate that arsenic is predominantly accumulated as As(III) in the leaves. XANES and EXAFS results show that As(III) in the leaves is primarily present as aqueous arsenite species. The plant actively maintains arsenic in this reduced oxidation state, because after sample collection and subsequent aging and drying of the plant material, As(III) is gradually oxidized to As(V). We think that these arsenite species are sequestered in vacuoles. At extremely high As concentrations (ca. 1% As per dry weight) arsenic in the fern leaves is coordinated to a significant degree by sulfur in addition to oxygen. This spectral signature indicates that thiol-rich compounds are implicated in the biochemical transformations of arsenic within the plant. PMID- 12636276 TI - Electrophoretic assembly of naturally occurring humic substances as thin films. AB - Polydisperse humic acid thin films on optically transparent electrodes (OTEs) have been prepared by electrophoretic deposition from a solution of Suwanee River humic acid (SHA) in ethanol/acetonitrile. The thickness of the film and the rate of deposition of SHA are dependent on the applied voltage and the concentration of the solution. Tapping-mode atomic force microscopy (TM-AFM) confirms the assembly of SHA aggregates on the electrode surface. The ability of these thin films to incorporate redox-active species such as ferrocene from solution is demonstrated by cyclic voltammetry experiments. A linear dependence of the peak current for the oxidation of ferrocene as a function of scan rate indicates that the ferrocene is incorporated into the humic membrane. PMID- 12636277 TI - Temperature dependence of the characteristic travel distance. AB - The effect of temperature variation on the environmental fate of organic chemicals can be evaluated in steady-state multimedia box models by expressing chemical partitioning data and reaction rate coefficients as functions of temperature. Using such a modelthetemperature dependence of the characteristic travel distance in air L(A), which is a measure for the atmospheric long-range transport potential of organic chemicals, is calculated. Simulations are reported for a set of 40 chemicals of environmental interest. Increasing temperature is shown to have two opposing effects on L(A). Rates of chemical transformations in the atmosphere (k(air)) and surface media are increased, which reduces L(A). Rates of atmospheric deposition (k(dep)) are reduced leading to increased mobility and L(A). Accordingly, L(A) can monotonically increase or decrease with increasing temperature, or it can have a maximum in the modeled temperature range, but it cannot have a minimum. For chemicals with a strong temperature dependence of k(air) relative to k(dep), L(A) will increase with increasing temperature. Results for selected polychlorinated biphenyls are compared to monitoring data yielding qualitative agreement when chemical properties are adjusted to mean temperatures for the measurement period. The results demonstrate that the temperature dependence of the characteristic travel distance is highly dependent on chemical characteristics and can be counterintuitive. The use of mass balance models is thus essential. The difference between the L(A) values at 5 degrees C and 30 degrees C can be up to a factor of 6. Accordingly, chemical ranking with respect to L(A) can change significantly if performed at different temperatures. Implications of the different temperature dependencies on long range transport to polar regions are discussed. PMID- 12636279 TI - Analysis of bacterial random motility in a porous medium using magnetic resonance imaging and immunomagnetic labeling. AB - In this study, we demonstrate the application of immunomagnetic labeling and magnetic resonance imaging (MRI) for the noninvasive visualization of changes in bacterial density distributions as a function of time in a water-saturated porous medium. Magnetite particles (50-60 nm diameter) were attached via a monoclonal antibody to the surface' of Escherichia coli K12 NR50 cells. The cells maintained their motility after labeling, and the presence of the magnetite did not significantly alter cell swimming speed. Diffusive migration for both motile and nonmotile E. coli through a porous medium with a particle-diameter distribution of 250-300 microm was compared. The movement of the nonmotile cells was described by an effective random motility coefficient consistent with Brownian diffusion of a nonmotile colloid. An effective coefficient determined a priori from bacterial motility in an aqueous medium and properties of the porous medium adequately described the movement of the motile cells. The ability to noninvasively visualize bacterial concentrations within an opaque porous medium in real time provides researchers with a powerful tool for studying bacterial transport in porous media. This is important for understanding the impact of bacterial transport on remediation strategies for environmental cleanup of polluted groundwater. PMID- 12636278 TI - Comparison of three techniques used to measure diffusive gas exchange from sheltered aquatic surfaces. AB - Three approaches commonly used to quantify diffusive gas exchange across aquatic surfaces were compared in a densely treed, low-wind environment Diffusive surface fluxes of carbon dioxide (CO2) and methane (CH4) from a small boreal reservoir were estimated using (i) surface water concentrations, the thin boundary layer (TBL) equation, and gas transfer velocities (k) calculated using sulfur hexafluoride (SF6); (ii) surface water concentrations, the TBL equation, and k estimated from wind speed; and (iii) static floating chambers (FCs). Comparisons were made during three different approximately 10-day intervals (August 2000, June and September 2001). CO2 and CH4 fluxes estimated from SF6-derived k were on average 1-3 times greater than those determined from wind-estimated k Overall agreement between FC CO2 and CH4 flux estimates and those based on SF6 and wind speed derived kvalues was much weaker, with FC CO2 and CH4 flux estimates ranging from -9 to 23 times those based on SF6 and wind-estimated k values. Chamber deployment likely enhanced gas transfer through disturbance of the surface boundary layer, and results of this study suggest that caution must be exercised concerning the use of FCs on very still water surfaces. Furthermore, findings of this study contradict the common belief that use of wind speed to approximate k is inappropriate for small bodies of water characterized by low winds and surface obstructions. PMID- 12636280 TI - Application of high-angle annular dark field scanning transmission electron microscopy, scanning transmission electron microscopy-energy dispersive X-ray spectrometry, and energy-filtered transmission electron microscopy to the characterization of nanoparticles in the environment. AB - A major challenge to the development of a fundamental understanding of transport and retardation mechanisms of trace metal contaminants (<10 ppm) is their identification and characterization at the nanoscale. Atomic-scale techniques, such as conventional transmission electron microscopy, although powerful, are limited by the extremely small amounts of material that are examined. However, recent advances in electron microscopy provide a number of new analytical techniques that expand its application in environmental studies, particularly those concerning heavy metals on airborne particulates or water-borne colloids. High-angle annular dark field scanning transmission electron microscopy (HAADF STEM), STEM-energy-dispersive X-ray spectrometry (EDX), and energy-filtered TEM (EFTEM) can be effectively used to identify and characterize nanoparticles. The image contrast in HAADF-STEM is strongly correlated to the atomic mass: heavier elements contribute to brighter contrast. Gold nanocrystals in pyrite and uranium nanocrystals in atmospheric aerosols have been identified by HAADF-STEM and STEM EDX mapping and subsequently characterized by high-resolution TEM (HRTEM). EFTEM was used to identify U and Fe nanocrystals embedded in an aluminosilicate. A rare, As-bearing nanophase, westerveldite (FeAs), was identified by STEM-EDX and HRTEM. The combined use of these techniques greatly expands the effective application of electron microscopy in environmental studies, especially when applied to metals of very low concentrations. This paper describes examples of how these electron microbeam techniques can be used in combination to characterize a low concentration of heavy metals (a few ppm) on nanoscale particles. PMID- 12636281 TI - In situ two-dimensional high-resolution profiling of sulfide in sediment interstitial waters. AB - A recently developed technique of diffusive gradients in thin films (D6T) computer-imaging densitometry (CID) was improved to study in situ two-dimensional distributions of sulfide in sediment interstitial waters adjacent to the DGT device. The in situ profiler accumulates sulfide from the sediment interstitial water through a diffusive gel onto a AgI binding gel to form black Ag2S. The amount of sulfide bound in the binding gel is then determined from the gray scale density of the dried binding gel. New gel-making procedures were employed to produce stable, evenly dispersed AgI binding gels and to minimize the darkening of the AgI gel upon exposure to light The improved DGT-CID technique was used to profile the distribution of sulfide in sediment interstitial waters at Delta Marsh, a highly productive Prairie wetland on the south shore of Lake Manitoba, at a vertical and lateral resolution of < or = 0.4 mm. The in situ high resolution microprofiles revealed unprecedented two-dimensional heterogeneity in sulfide concentrations in the sediment interstitial waters adjacent to the DGT device. The mosaic distribution of oxic and sulfidic microenvironments suggested not only the complexity and heterogeneity of the biogeochemistry of sulfur species and sulfide-binding metals (e.g., Cd, Cu, Pb, Hg, Zn) in sediments but also the capability of aquatic organisms for coping with the sulfidic environment PMID- 12636282 TI - Monitoring oxidation of chlorinated ethenes by permanganate in groundwater using stable isotopes: laboratory and field studies. AB - Permanganate injection is increasingly applied for in situ destruction of chlorinated ethenes in groundwater. This laboratory and field study demonstrates the roles that carbon isotope analysis can play in the assessment of oxidation of trichloroethene (TCE) by permanganate. In laboratory experiments a strong carbon isotope fractionation was observed during oxidation of TCE with similar isotopic enrichment factors (-25.1 to -26.8 per thousand) for initial KMnO4 concentrations between 67 and 1,250 mg/L. At the field site, a single permanganate injection episode was conducted in a sandy aquifer contaminated with TCE as dense nonaqueous liquid (DNAPL). After injection, enriched delta13C values of up to +204% and elevated Cl- concentrations were observed at distances of up to 4 m from the injection point. Farther away, the Cl- increased without any change in delta13C of TCE suggesting that Cl- was not produced locally but migrated to the sampling point Except for the closest sampling location to the injection point, the delta13C rebounded to the initial 613C again, likely due to dissolution of DNAPL Isotope mass balance calculations made it possible to identify zones where TCE oxidation continued to occur during the rebound phase. The study indicates that delta13C values can be used to assess the dynamics between TCE oxidation and dissolution and to locate zones of oxidation of chlorinated ethenes that cannot be identified from the Cl- distribution alone. PMID- 12636283 TI - Enhancing phenanthrene biomineralization in a polluted soil using gaseous toluene as a cosubstrate. AB - Laboratory experiments were conducted to study the potential of adding gaseous toluene, as a readily degradable carbon source, to enhance phenanthrene mineralization in polluted soil (1,000 mg/kg(dry soil)) aged for 400 days. Experiments were conducted in 0.5-L column reactors packed with a mixture of (80:20 w(wet)/w(wet)) spiked soil and vermiculite and fed with 1 g m(-3)reactor h(-1) toluene load in air. Removal efficiencies of 100% for toluene and greater than 95% for phenanthrene were obtained in 190 h. Evolved CO2 showed that phenanthrene mineralization increased from 39% to 86% in columns treated with gaseous toluene. Phthalic acid was identified as the principal soluble intermediate, which accumulated when no toluene was added. Increased phenanthrene uptake and mineralization with toluene can be attributed to increased biomass and the induction of enzymes involved in the intermediate mineralization. In microcosm experiments, phthalic acid mineralization increased from 19% to 81% within 50 h in the presence of toluene. Experiments with 14C-labeled phenanthrene confirmed the enhancement of phenanthrene mineralization from 45% to 83% in 385 h with toluene as a second carbon source. The results indicate thatthe addition of an appropriate gaseous cosubstrate could be an adequate strategy to enhance mineralization of PAHs in soil. PMID- 12636284 TI - Carbohydrate-derived chlorinated compounds in ECF bleaching of hardwood pulps: formation, degradation, and contribution to AOX in a bleached kraft pulp mill. AB - Five monochlorinated compounds derived from glucuronoxylan were identified in the liquid process streams of a kraft pulp mill producing hardwood pulp with ECF bleaching, representing ca. 15-20% of the AOX of the bleaching filtrates. The environmental risk of such compounds is negligible because a major fraction (70 80%) is degraded during effluent mixing and neutralization, and about 20-30% is degraded during the biological treatment of the mixed effluent. Only less than 3.5% (0.009-0.017 kg/ tAD) of the compounds formed in the bleaching leave the mill in the final effluent. PMID- 12636285 TI - Isolation, selection, and characterization of lactic acid bacteria for a competitive exclusion product to reduce shedding of Escherichia coli O157:H7 in cattle. AB - Lactic acid bacteria (LAB) were selected on the basis of characteristics indicating that they would be good candidates for a competitive exclusion product (CEP) that would inhibit Escherichia coli O157:H7 in the intestinal tract of live cattle. Fecal samples from cattle that were culture negative for E. coli O157:H7 were collected. LAB were isolated from cattle feces by repeated plating on deMan Rogosa Sharpe agar and lactobacillus selection agar. Six hundred eighty-six pure colonies were isolated, and an agar spot test was used to test each isolate for its inhibition of a four-strain mixture of E. coli O157:H7. Three hundred fifty five isolates (52%) showed significant inhibition. Seventy-five isolates showing maximum inhibition were screened for acid and bile tolerance. Most isolates were tolerant of acid at pH levels of 2, 4, 5, and 7 and at bile levels of 0.05, 0.15, and 0.3% (oxgall) and were subsequently identified with the API system. Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus delbreukii, Lactobacillus salivarius, Lactobacillus brevis, Lactobacillus cellobiosus, Leuconostoc spp., and Pediococcus acidilactici were the most commonly identified LAB. Nineteen strains were further tested for antibiotic resistance and inhibition of E. coli O157:H7 in manure and rumen fluid. Four of these 19 strains showed susceptibility to all of the antibiotics, 13 significantly reduced E. coli counts in manure, and 15 significantly reduced E. coli counts in rumen fluid (P < 0.05) during at least one of the sampling periods. One of the strains, M35, was selected as the best candidate for a CEP. A 16S rRNA sequence analysis of M35 revealed its close homology to Lactobacillus crispatus. The CEP developed will be used in cattle-feeding trials. PMID- 12636286 TI - Molecular characterization of Arcobacter isolates collected in a poultry slaughterhouse. AB - In a poultry slaughterhouse, Arcobacter contamination was examined over a period of 1 week to establish possible routes of contamination. Samples were collected from the slaughter equipment and from processing water before the onset of slaughter and from the first broiler flock slaughtered on each sampling day. Characterization of 1,079 isolates by enterobacterial repetitive intergenic consensus-polymerase chain reaction and a random amplified polymorphic DNA assay resulted in the delineation of 159 Arcobacter butzleri and 139 Arcobacter cryaerophilus types. From almost all 140 neck skin samples collected before and after evisceration, A. butzleri and A. cryaerophilus were isolated simultaneously at contamination levels ranging from 10(1) to 10(4) CFU/g. Only six A. butzleri types present in the slaughterhouse environment were also present on the broiler carcasses. None of the A. cryaerophilus genotypes were detected in both the neck skin and the environmental samples. All A. butzleri types isolated from the feather samples were also isolated from broiler neck skin samples. The slaughter equipment was contaminated with arcobacters before the onset of slaughter, but it appeared unlikely that contamination through the slaughter equipment alone explained the high contamination levels on poultry products. Arcobacters were also present in processing water, but types present in water and poultry products were different. Characterization of the Arcobacter isolates did not clarify the routes of transmission, probably because of the extreme heterogeneity among Arcobacter isolates. However, the results obtained in this study brought to light insufficient decontamination at the processing plant involved in the study and confirmed the survival capacity of certain A. butzleri strains. PMID- 12636287 TI - Enterobacter sakazakii: infectivity and enterotoxin production in vitro and in vivo. AB - Enterobacter sakazakii has been implicated as the causal organism in a severe form of neonatal meningitis, with reported mortality rates of 40 to 80%. Dried infant formula has been identified as a potential source of the organism in both outbreaks and sporadic cases. In this study, clinical and foodborne isolates of E. sakazakii were evaluated for enterotoxin production by the suckling mouse assay. In addition, suckling mice were challenged both orally and by intraperitoneal injection. Of 18 E. sakazakii strains evaluated, four were found to test positive for enterotoxin production. All strains of E. sakazakii were lethal to suckling mice at 10(8) CFU per mouse by intraperitoneal injection, while two strains caused death by the peroral route. In in vitro assays, CHO, Vero, and Y-1 cells demonstrated both cell lysis and rounding when exposed to E. sakazakii strain LA filtrates. This is the first report describing any putative virulence factors of E. sakazakii. PMID- 12636288 TI - Control of Clostridium perfringens germination and outgrowth by buffered sodium citrate during chilling of roast beef and injected pork. AB - Inhibition of the germination and outgrowth of Clostridium perfringens by buffered sodium citrate (Ional) and buffered sodium citrate supplemented with sodium diacetate (Ional Plus) during the abusive chilling of roast beef and injected pork was evaluated. Beef top rounds or pork loins were injected with a brine containing NaCl, potato starch, and potassium tetrapyrophosphate to yield final in-product concentrations of 0.85, 0.25, and 0.20%, respectively. Products were ground and mixed with Ional or Ional Plus at 0, 0.5, 1.0, and 2.0%. Each product was mixed with a three-strain C. perfringens spore cocktail to obtain final spore concentrations of ca. 2.5 log10 spores per g. Chilling of roast beef from 54.4 to 7.2 degrees C resulted in C. perfringens population increases of 1.51 and 5.27 log10 CFU/g for 18- and 21-h exponential chill rates, respectively, while chilling of injected pork resulted in increases of 3.70 and 4.41 log10 CFU/g. The incorporation of Ional into the roast beef formulation resulted in C. perfringens population reductions of 0.98, 1.87, and 2.47 log10 CFU/g with 0.5, 1.0, and 2.0% Ional, respectively, over 18 h of chilling, while > or = 1.0% Ional Plus was required to achieve similar reductions (reductions of 0.91 and 2.07 log10 CFU/g were obtained with 1.0 and 2.0% Ional Plus, respectively). An Ional or Ional Plus concentration of > or = 1.0% was required to reduce C. perfringens populations in roast beef or injected pork chilled from 54.4 to 7.2 degrees C in 21 h. Cooling times for roast beef or injected pork products after heat processing can be extended to 21 h through the incorporation of > or = 1.0% Ional or Ional Plus into the formulation to reduce the potential risk of C. perfringens germination and outgrowth. PMID- 12636289 TI - Viability of Clostridium perfringens, Escherichia coli, and Listeria monocytogenes surviving mild heat or aqueous ozone treatment on beef followed by heat, alkali, or salt stress. AB - The threat of pathogen survival following ozone treatment of meat necessitates careful evaluation of the microorganisms surviving under such circumstances. The objective of this study was to determine whether sublethal aqueous ozone treatment (3 ppm of O3 for 5 min) of microorganisms on beef surfaces would result in increased or decreased survival with respect to subsequent heat, alkali, or NaCl stress. A mild heat treatment (55 degrees C for 30 min) was used for comparison. Reductions in three-strain cocktails of Clostridium perfringens, Escherichia coli O157:H7, and Listeria monocytogenes on beef following the heat treatment were 0.14, 0.77, and 1.47 log10 CFU/g, respectively, whereas reductions following ozone treatment were 1.28, 0.85, and 1.09 log10 CFU/g, respectively. C. perfringens cells exhibited elevated heat resistance at 60 degrees C (D60 [time at 60 degrees C required to reduce the viable cell population by 1 log10 units or 90%] = 17.76 min) following heat treatment of beef (55 degrees C for 30 min) but exhibited reduced viability at 60 degrees C following ozone treatment (D60 = 7.64 min) compared with the viability of untreated control cells (D60 = 13.84 min). The D60-values for L. monocytogenes and E. coli O157:H7 following heat and ozone exposures were not significantly different (P > 0.05). C. perfringens cells that survived ozone treatment did not exhibit increased resistance to pH (pH 6 to 12) relative to non-ozone-treated cells when grown at 37 degrees C for 24 h. The heat treatment also resulted in decreased numbers of surviving cells above and below neutral pH values for both E. coli O157:H7 and L. monocytogenes relative to those of non-heat-treated cells grown at 37 degrees C for 24 h. There were significant differences (P < 0.05) in C. perfringens reductions with increasing NaCl concentrations. The effects of NaCl were less apparent for E. coli and L. monocytogenes survivors. It is concluded that pathogens surviving ozone treatment of beef are less likely to endanger food safety than are those surviving sublethal heat treatments. PMID- 12636290 TI - A novel method for the reduction of numbers of Listeria monocytogenes cells by freezing in combination with an essential oil in bacteriological media. AB - The use of multiple freeze (-20 degrees C)-thaw cycles in combination with isoeugenol and polysorbate 80 was investigated as a method for the reduction of numbers of Listeria monocytogenes cells in a bacteriological medium. Three freeze (1 h, -20 degrees C)-thaw cycles in the presence of isoeugenol at concentrations of 0, 100, and 300 ppm resulted in average L. monocytogenes reductions of 0.69, 2.65, and 3.3 log10 MPN (most probable number) per ml, respectively. Increasing the number of freeze-thaw cycles further decreased cell numbers, with reductions of nearly 5 log10 MPN/ml being obtained with six freeze-thaw cycles. Freeze-thaw cycles were effective in reducing cell numbers at isoeugenol concentrations down to 25 ppm. Rapid freezing rates with liquid nitrogen were found to be less effective in reducing numbers of L. monocytogenes cells. Two rapid freeze-thaw cycles in the presence of 100 ppm isoeugenol and polysorbate 80 resulted in a reduction of 1.45 log10 MPN/ml. Two freezing (-20 degrees C) cycles involving slow freezing and thawing rates with samples being held frozen for 6 h for each cycle resulted in reductions larger than those obtained with faster freezing rates. It was found that complete thawing in freeze-thaw cycles was not necessary to achieve bactericidal action. The application of multiple freeze-thaw cycles in combination with low concentrations of isoeugenol could effectively reduce numbers of L. monocytogenes cells in bacteriological media. PMID- 12636291 TI - Effects of acid adaptation and modified marinades on survival of postdrying Salmonella contamination on beef jerky during storage. AB - This study was undertaken to evaluate the survival of acid-adapted and nonadapted Salmonella cultures inoculated after drying on beef jerky that had been treated with marinades before drying at 60 degrees C for 10 h. Beef slices were (i) not treated prior to refrigeration at 4 degrees C for 24 h (control [C]); (ii) marinated with traditional marinade (TM), (iii) marinated with TM modified with 1.2% sodium lactate, 9% acetic acid, and 68% soy sauce containing 5% ethanol (MM) at twice the amount used in the TM treatment; (iv) dipped into 5% acetic acid and then marinated with TM (AATM); and (v) dipped into 1% Tween 20, then dipped into 5% acetic acid, and then marinated with TM (TWTM); after each treatment, meat slices were refrigerated at 4 degrees C for 24 h prior to drying. Dried slices were inoculated with acid-adapted or nonadapted Salmonella (ca. 5.7 log CFU/cm2) prior to aerobic storage at 25 degrees C for 60 days. Tryptic soy agar with 0.1% pyruvate, as well as xylose-lysine-tergitol 4 (XLT4) agar, was used to determine survivor counts. Bacterial decreases achieved with the different treatments were found to be in the following order: TWTM (5.4 to 6.3 log units) > or = AATM > or = MM > C > or = TM (2.9 to 5.1 log units). Acid-adapted Salmonella decreased faster than nonadapted Salmonella for all treatments. Bacterial populations decreased to below the detection limit (-0.4 log CFU/cm2) in as few as 14 days or remained detectable by direct plating after 60 days of storage, depending on acid adaptation, treatment, and agar media. The results of this study indicate that the modified marinades used in jerky processing and the low water activity of the dried product provide antimicrobial effects against possible postprocessing contamination with Salmonella, while the preparation of cultures under acid adaptation conditions did not increase Salmonella survival during storage and may have reduced it. PMID- 12636292 TI - Microbiological study of ready-to-eat salad vegetables from retail establishments uncovers a national outbreak of salmonellosis. AB - The increasing availability of bagged prepared salad vegetables reflects consumer demand for fresh, healthy, convenient, and additive-free foods that are safe and nutritious. During May and June 2001 a study of retail bagged prepared ready-to eat salad vegetables was undertaken to determine the microbiological quality of these vegetables. Examination of the salad vegetables revealed that the vast majority (3,826 of 3,852 samples; 99.3%) were of satisfactory or acceptable microbiological quality according to Public Health Laboratory Service microbiological guidelines, while 20 (0.5%) samples were of unsatisfactory microbiological quality. Unsatisfactory quality was due to Escherichia coli and Listeria spp. (not Listeria monocytogenes) levels in excess of 10(2) CFU/g. However, six (0.2%) samples were of unacceptable microbiological quality because of the presence of Salmonella (Salmonella Newport PT33 [one sample], Salmonella Umbilo [three samples], and Salmonella Durban [one sample]) or because of a L. monocytogenes level of 660 CFU/g, which indicates a health risk. In each case, the retailer involved and the UK Food Standards Agency were immediately informed, and full investigations were undertaken. Nineteen cases of Salmonella Newport PT33 infection were subsequently identified throughout England and Wales. The outbreak strain of Salmonella Newport PT33 isolated from the salad and from humans had a unique plasmid profile. Campylobacter spp. and E. coli O157 were not detected in any of the samples examined. The presence of Salmonella, as well as high levels of L. monocytogenes, is unacceptable. However, minimally processed cut and packaged salad is exposed to a range of conditions during growth, harvest, preparation, and distribution, and it is possible that these conditions may increase the potential for microbial contamination, highlighting the necessity for the implementation of good hygiene practices from farm to fork to prevent contamination and/or bacterial growth in these salad products. PMID- 12636293 TI - Effects of herbal essential oils used to extend the shelf life of freshwater reared Asian sea bass fish (Lates calcarifer). AB - Sensory and microbiological characteristics of Asian sea bass (Lates calcarifer) fish raised in a freshwater pond were evaluated during cold storage for 33 days. Whole fish (averaging 400 g each) were stored in a cold storage room at 0 to 2 degrees C. Essential oils of herbs--thyme (Thymus vulgaris) and oregano (Origanum vulgare)--added at 0.05% (vol/vol) were used as preservatives. On the basis of the sensory test results as well as the bacteriological tests, the addition of oregano and thyme essential oils can considerably slow the process of spoilage. The fish treated with these oils were still fit for human consumption after 33 days of storage. The results obtained through sensory tests are corroborated to a great extent by the chemical (hypoxanthine) tests and to a lesser extent by the physical (Cosmos units) tests. The initial total bacteriological counts were 1.7 x 10(3) CFU/cm2 on the fish surface and <10(2) CFU/g in the fish flesh, and in the control treatment (without preservatives), these counts rose continuously, reaching around 10(7) CFU/cm2 on the fish surface and 10(3) CFU/g in the flesh after 33 days of storage at 0 to 2 degrees C. The use of herbal essential oils as preservatives, on the other hand, resulted in a maximal count of 10(5) CFU/cm2 on the fish surface, while the bacterial count in the flesh remained <10(2) CFU/g by the end of the 33-day storage period at 0 to 2 degrees C. PMID- 12636294 TI - Antagonistic action of cells of Lactobacillus delbrueckii subsp. lactis against pathogenic and spoilage microorganisms in fresh meat systems. AB - Cells of Lactobacillus delbrueckii subsp. lactis RM2-5 were added to various meat model systems that had been inoculated with Escherichia coli O157:H7 or Salmonella Typhimurium to determine whether these lactobacilli were antagonistic to the pathogens during storage at 5 degrees C. Experiments in which L. delbrueckii subsp. lactis RM2-5 was directly applied to the surfaces of beefsteaks resulted in significant (P < 0.05) reductions in the growth of psychrotrophs and coliforms plus a slight decrease in the numbers of E. coli O157:H7 over time relative to those for control samples to which no lactobacilli had been added. Experiments involving the direct application of L. delbrueckii subsp. lactis RM2-5 to the surfaces of freshly slaughtered beef and pork carcass samples inoculated with either E. coli O157:H7 or Salmonella Typhimurium showed significant (P < 0.05) declines in numbers of the pathogens as well as a reduction in the growth of psychrotrophs during storage at 5 degrees C for 6 days. The results of the experiments suggest that lactobacillus cultures have potential for use in an intervention technology for the control of foodborne pathogens, especially on the surfaces of beef and pork carcasses. The results of this study also suggest that an extension of the shelf life of meat can result from the decreased growth of psychrotrophic spoilage organisms. PMID- 12636295 TI - Kinetics of adsorption and desorption of aflatoxin B1 by viable and nonviable bacteria. AB - The reactions involved in the binding (adsorption) and release (desorption) of aflatoxin B1 (AFB1) to and from the surface of bacteria were investigated. Viable and heat-killed Lactobacillus rhamnosus GG, L. rhamnosus LC-705, and Propionibacterium freudenreichii subsp. shermanii JS were incubated in phosphate buffered saline containing variable concentrations (0.0017 to 13.3 microg/ml) of AFB1. The relationship between the bacterial surface hydrophobicity and the AFB1 adsorption affinity was also investigated. A linear relationship was observed between the specific rate of AFB1 adsorption and the AFB1 concentration for all bacteria. The nature of desorption of adsorbed AFB1 was investigated by repetitive aqueous washes. A linear relationship was observed between the natural log value of the concentration of AFB1 adsorbed and the number of washes for all bacteria studied. The desorption constants were strain-dependent and were lower for heat-killed bacteria than for viable bacteria. Heat treatment appears to alter the surface properties of the bacteria rather than expose new adsorption sites. No correlation was found between the hydrophobicity and the AFB1 adsorption affinity. PMID- 12636297 TI - Detection of Listeria monocytogenes in salmon using the Probelia polymerase chain reaction system. AB - A validation was conducted on the performance of a commercially available polymerase chain reaction (PCR) kit (Probelia) in comparison with International Organization for Standardization (ISO) method 11290-1 (adopted as an Australian New Zealand Standard Method, AS/NZS 1766.2.16.1:1998) for the detection of Listeria monocytogenes in salmon samples. The validation was conducted following the guidelines of an Australian New Zealand Standard (Guide to Determining the Equivalence of Food Microbiology Test Methods, Part 1, Qualitative Tests, AS/NZS 4659.1:1999), which adopts an approach similar to that recommended by the Association of Analytical Communities Microbiology Method Validation Program for Performance Tested and Peer Verified Methods. The validation study involved the use of five cultures of L. monocytogenes, each challenged at a single level of inoculation into five different types of salmon samples. A total of 60 salmon samples (30 unchallenged and 30 challenged) were tested using both the PCR method and the ISO method. Results from this study indicated that the Probelia PCR method is equivalent to the ISO method. In addition, the detection sensitivity of the Probelia PCR system was determined as approximately 0.5 CFU per PCR assay (equivalent to 20 CFU/ml broth culture) for a pure culture of L. monocytogenes. The Probelia PCR method offers the advantage of detecting L. monocytogenes to genetic specificity within 48 to 50 h, whereas the ISO method requires 5 days for negative results with additional days for confirmed positive results by the use of other biochemical and cultural tests. PMID- 12636296 TI - Reduction of aflatoxins by Korean soybean paste and its effect on cytotoxicity and reproductive toxicity--part 2. Antigenotoxic effects of the methanol extract of Korean soybean paste (doen-jang) on aflatoxin B1-induced bacterial reverse mutation and chromosome aberration. AB - A bacterial reverse-mutation assay with Salmonella Typhimurium TA1535, TA1537, TA98, TA100, and TA102 and an in vitro chromosome aberration assay with Chinese hamster lung (CHL) cells were used to investigate the genotoxicity of the methanol extract of Korean soybean paste (doen-jang [dwen-jahng]) and its antigenotoxic activity against aflatoxin B1. The methanol extract revealed nonmutagenic potential for all of the bacterial strains tested. The extract significantly reduced the numbers of revertants per plate when it was added to the assay system with Salmonella Typhimurium TA100 (P < 0.05). The extract also exhibited significant inhibitory effects on chromosome aberration in CHL cells (P < 0.05). The findings of this work indicate that the methanol extract of Korean soybean paste could have strong potential as an antigenotoxic material. PMID- 12636298 TI - Evaluation and interlaboratory validation of a selective agar for phosphatidylinositol-specific phospholipase C activity using a chromogenic substrate to detect Listeria monocytogenes from foods. AB - Phosphatidylinositol-specific phospholipase C (PI-PLC) activity is a potential virulence factor and is exhibited only by the Listeria species Listeria monocytogenes and Listeria ivanovii. A chromogenic substrate for the direct detection of PI-PLC activity is available in a new medium (BCM L. monocytogenes plating agar). The use of a chromogenic substrate offers a mechanism with which to directly screen for L. monocytogenes and L. ivanovii other than the esculin used in Oxford (OXF) and Palcam (PAL) agars, which screen for all Listeria species. The specificity levels of BCM plating agar and of BCM confirmation and rhamnose agars were evaluated with 107 Listeria and 10 Bacillus species isolates. In addition, BCM L. monocytogenes plating agar was compared with standard Listeria selective agars (OXF and PAL agars) with regard to the recovery of L. monocytogenes from 2,000 food and environmental samples obtained from eight participating laboratories. A Listeria species was isolated from at least one of the agars in 209 analyses, and L. monocytogenes was isolated in 135 of these analyses. In 27 of the analyses in which L. monocytogenes was isolated, one or more of the selective differential agars used failed to isolate L. monocytogenes, and therefore the results of these analyses were discrepant. Relative to a reference method involving the use of all three agars (OXF, PAL, and BCM agars), the OXF-BCM, PAL-BCM, and OXF-PAL combinations had sensitivities of 99.3, 99.2, and 90.2%, respectively. In statistical analyses of the different combinations of agars, the OXF-BCM and BCM-PAL combinations were found to be superior to the OXF PAL combination for the detection of L. monocytogenes. PMID- 12636299 TI - Recovery of Salmonella from retail broilers by a whole-carcass enrichment procedure. AB - Fresh whole broiler carcasses were purchased from grocery stores over a 20-week period. Carcasses were selected on the basis of their having intact packages and unique U.S. Department of Agriculture (USDA) plant numbers and sell-by dates, such that each bird represented a single processing plant-processing day combination. Carcasses were tested for Salmonella with a rinse aliquot obtained after whole-bird incubation in the rinse media for 24 h. On the basis of the number of unique processing plants (USDA plant numbers) and expiration dates involved, the number of birds available each week ranged from 6 to 17. Over the 20-week period, 251 independent carcasses from 14 processing plants were tested. The percentages of carcasses testing positive for Salmonella ranged from 0 (for 1 week) to >60% (for 3 weeks). For only 4 of the 20 weeks was an incidence of Salmonella-positive carcasses of <20% found. For the entire 20-week study, 85 (33.9%) of the 251 carcasses tested were found to be Salmonella positive. For those processing plants from which >10 carcasses were obtained, the percentages of carcasses testing positive for Salmonella ranged from <20 (two plants) to >40% (four plants). These results indicate that a whole-carcass enrichment may be more sensitive for the detection of Salmonella-positive carcasses than the traditional whole-carcass rinse followed by immediate testing of a subsample aliquot when small numbers of Salmonella are expected. PMID- 12636300 TI - Immunological detection of Fusarium species in cornmeal. AB - An indirect enzyme-linked immunosorbent assay (ELISA) was developed to detect Fusarium species in foods. Antibodies to proteins extracted from the mycelia of Fusarium graminearum and Fusarium moniliforme (verticillioides) were produced in New Zealand white rabbits. These antibodies detected 13 Fusarium species in addition to the producer strains. Levels of Fusarium semitectum and Fusarium tricinctum strains were below the detection threshold. The specificity of the assay was tested against 70 molds and yeasts belonging to 23 genera. One strain of Monascus species and one strain of Phoma exigua were detected; however, these two molds are not common contaminants of cereal grains or foods and should not interfere with the assay. The indirect ELISA's detection limits for F. graminearum and F. moniliforme were 0.1 and 1 microg of mold mycelium per ml of a cornmeal mixture, respectively. When spores of each mold were added individually to cornmeal mixtures (at ca. 10 spores per g) and incubated at 25 degrees C, these spores were detected by the indirect ELISA when they reached levels of 10(2) to 10(3) CFU/ml after 24 to 36 h. The indirect ELISA developed here shows promise for the detection of Fusarium species in grains or foods. PMID- 12636301 TI - Prophylactic feeding of Lactobacillus acidophilus NCFM to mice attenuates overt colonic hyperplasia. AB - The objective of this project was to determine if the probiotic Lactobacillus acidophilus NCFM would protect mice from developing transmissible murine colonic hyperplasia (TMCH) caused by Citrobacter rodentium. Our hypothesis was that the oral administration of L. acidophilus NCFM to mice would mitigate colonic hyperplasia and modulate the host immune response. A concurrent administration (CA) study was performed by feeding mice phosphate-buffered saline (PBS), C. rodentium only, L. acidophilus NCFM only, or C. rodentium and NCFM concurrently on the same day. The mice in the CA study were not protected by the probiotic, since their mean colon sample weights (0.109 g) were significantly higher than those of the PBS controls (0.0774 g), and the hematoxylin and eosin-stained samples showed histological changes typically associated with TMCH. A prophylactic feeding (PF) study was performed by orally feeding mice PBS or NCFM once daily for 20 consecutive days; in addition, on day 7, mice were challenged with either PBS or C. rodentium. Mice in the PF study were protected when they consumed the probiotic prior to the pathogen challenge, since their mean colon sample weights (0.0812 g) were not significantly higher than those of the controls (0.0753 g). The hematoxylin and eosin-stained samples appeared similar to the control samples, and the intestinal interleukin (IL)-15 and gamma interferon (IFN-gamma) mRNA levels were reduced. L. acidophilus NCFM did attenuate overt colonic hyperplasia when fed to mice prior to challenge with C. rodentium. The mouse model used in this study enabled us to investigate the efficacy of the L. acidophilus NCFM in preventing gastrointestinal disease and is a valid model for future probiotic research. PMID- 12636302 TI - Interactions of lactic acid bacteria with human intestinal epithelial cells: effects on cytokine production. AB - As a participant in the mucosal immune response, the intestinal epithelial cell must respond to a variety of stimuli, including lactic acid bacteria (LAB) consumed in the diet. The objective of this study was to compare the abilities of several strains of LAB to modulate cytokine secretion by human intestinal epithelial cell (IEC) line HT-29. Certain strains of Lactobacillus rhamnosus, Lactobacillus delbrueckii, and Lactobacillus acidophilus suppressed the production of the chemokine RANTES by stimulated HT-29 IEC, although the magnitude of this suppression varied depending on the nature of the bacterial growth medium. Similarly, specific strains showed growth condition-dependent suppression of HT-29 interleukin-8 (IL-8) production. Strain-dependent effects were also seen for the suppression of tumor necrosis factor alpha (TNF-alpha) and transforming growth factor beta (TGF-beta) production. The binding of several of these bacterial strains to the HT-29 cell line was also examined. Different strains were found to have differing abilities to interact with IEC, with L. rhamnosus R0011 being the strain that generally had the most extensive effects on HT-29 cytokine production and also bound to HT-29 IEC most effectively. Modulation of IEC cytokine production has the potential to profoundly affect the mucosal microenvironment, influencing the immune response to pathogens and other ingested antigens. PMID- 12636303 TI - Accuracy of BRT and Delvotest microbial inhibition tests as affected by composition of ewe's milk. AB - The presence of drug residues in ewe's milk samples can be determined by microbial assays. The main limitation of these tests is the large number of false positive results associated with them. False-positive results can be explained by the interaction of certain substances naturally existing in ewe's milk with the growth of the microorganism used in the test. In this study, milk chemical composition (fat, protein, lactose, total solids), somatic cell counts (SCCs), free fatty acid concentrations, and lactoperoxidase system components were determined in order to investigate their influence on the rate of false-positive results for the BRT and Delvotest microbiological inhibitor tests. Milk samples were obtained after morning milking of Manchega ewes at 15, 30, 45, 60, 75, 90, 105, 120, and 135 days after parturition. The animals did not receive any kind of treatment or medicated feed throughout the experiment. The false-positive rates for BRT and Delvotest were 3.75 and 2.4%, respectively. When the logistic regression model was applied, the percentages of total solids for positive samples were significantly different from those for negative samples (16.90 versus 18.42% for BRT, 16.05 versus 18.45% for Delvotest), while the SCC logarithmic transformation was significantly higher for the positive samples than for the negative samples (5.38 versus 5.11 log units for BRT, 5.32 versus 5.11 log units for Delvotest). Moreover, Delvotest-positive samples exhibited thiocyanate concentrations higher than those of Delvotest-negative samples (8.18 mg/liter versus 6.85 mg/liter). Further analyses are needed to confirm the possible presence of antimicrobial residues in this particular type of milk sample. PMID- 12636304 TI - Levels of polychlorinated biphenyls in foods from Catalonia, Spain: estimated dietary intake. AB - From June to August 2000, food samples were randomly acquired in seven cities in Catalonia, Spain. Polychlorinated biphenyl (PCB) concentrations were determined for 108 samples of vegetables, fruits, pulses, cereals, fish and shellfish, meats and meat products, eggs, milk and dairy products, and oils and fats. Levels of 11 PCB congeners (IUPAC 28, 52, 77, 101, 105, 118, 126, 138, 153, 169, and 180) were determined by high-resolution gas chromatography-high-resolution mass spectrometry. For toxic equivalent (TEQ) calculations, World Health Organization (WHO) toxicity equivalent factors (WHO-TEFs) were used. The highest levels of most congeners were found in fish and shellfish (11,864.18 ng/kg [wet weight]), and the next highest levels, which were substantially lower, were found in milk and dairy products (674.50 ng/kg [wet weight]). For the general population of Catalonia, the total dietary intake of PCBs was found to be 150.13 pg WHO TEQ/day. The largest contribution to this intake came from fish and shellfish (82.87 pg WHO-TEQ/day) and dairy products (29.38 pg WHO-TEQ per day). A relatively large contribution was also noted for cereals (11.36 pg WHO-TEQ/day). Among the PCB congeners determined in this study, PCB 126 showed the largest contribution to total TEQ intake (50.56%). The data obtained in this study should be useful in risk assessment with regard to human PCB exposure through food in Catalonia. PMID- 12636305 TI - Implementation and analysis of an on-farm food safety program for the production of greenhouse vegetables. AB - Fresh fruits and vegetables are increasingly being recognized as vectors for foodborne illness. In 1998, an on-farm food safety program was developed for the Ontario Greenhouse Vegetable Growers (OGVG) in Ontario, Canada. This hazard analysis critical control point-based system was designed to reduce the potential of microbial contamination along the entire production and distribution process. A previous report (D. A. Powell, M. Bobadilla-Ruiz, A. Whitfield, M. W. Griffiths, and A. Luedtke, J. Food Prot. 65:918-923) outlined the first 2 years of the program; the present report documents the third year of the program's implementation. Through individual on-site grower visits and the microbiological testing of produce and water, the program's OGVG member requirements were determined, and the program's success was assessed. Growers' knowledge, understanding, and awareness of microbial risks associated with fresh produce have increased, and improvements in practices used in greenhouses and packing sheds were noted. PMID- 12636306 TI - Comparison of three commercial competitive-exclusion products for controlling Salmonella colonization of broilers in Brazil. AB - In this study, a new competitive-exclusion (CE) product, Mucosal Starter Culture (MSC), was compared with two other CE products (Aviguard and Avifree) commercially available in Brazil to evaluate their ability to protect newly hatched chicks against colonization by a strain of Salmonella Kedougou. This study was based on a previously published and recommended method for such products. Separate groups of the chicks were dosed orally with the respective treatment materials and challenged 24 h later, and their ceca were examined for Salmonella 5 days after challenge. Under the test conditions, only MSC and Aviguard gave statistically significant (P < 0.05) protection to the chicks, but the MSC treatment yielded the lowest mean level of cecal carriage and the smallest proportion of Salmonella-positive birds. PMID- 12636307 TI - Microbiological effects of hand washing at a beef carcass-breaking facility. AB - The hands of workers in the carcass-breaking facility at a beef packing plant were sampled by rinsing. Total aerobes, coliforms, and Escherichia coli were enumerated for each sample. The numbers of bacteria recovered from duplicate groups of 25 hand samples collected before and after hands were washed with an antibacterial gel, rinsed in a disinfectant solution, washed with the gel and rinsed with the disinfectant, or washed in the disinfectant for 20 s were similar for samples collected before work began after breaks. The numbers of bacteria recovered from samples collected before and after hands were washed with the antibacterial gel and rinsed in the disinfectant solution were similar for samples collected after work as well. However, the mean numbers of aerobes recovered from the four groups of hand samples after work were all >6.5 log CFU per hand, while 9 of the 10 corresponding values for groups of hand samples collected before work were <6.5 log CFU per hand; the total numbers of coliforms recovered from three groups of hand samples collected after work were >4 log CFU/25 hands, while 9 of the corresponding values for groups of hand samples collected before work were <4 log CFU/25 hands. The total numbers of E. coli recovered from all groups of hand samples collected after work were >3.5 log CFU/25 hands, while 9 of the corresponding values for groups of hand samples collected before work were <3 log CFU/25 hands. Thus, although washing and/or rinsing apparently did not reduce the numbers of bacteria on hands, fewer bacteria were recovered from hands before than after work. PMID- 12636308 TI - Behavior of enteroaggregative Escherichia coli in bottled spring and mineral water. AB - The ability of enteroaggregative Escherichia coli (EAEC) to survive in bottled mineral and spring water at common storage temperatures was investigated. Filtered mineral and spring waters were inoculated with EAEC (ca. 10(4) CFU/ml) and stored at 4, 10, and 23 degrees C. Water samples were analyzed every 3 days for viable EAEC by plating on tryptic soy agar plates over 60 days of storage. EAEC survived for the duration of the study in both mineral and spring waters. EAEC survival levels were significantly higher (P < 0.01) at 23 and 10 degrees C than at 4 degrees C. Furthermore, EAEC survival levels were significantly higher (P < 0.01) in mineral water than in spring water at 4 and 10 degrees C. The results of this study indicate that EAEC can survive in bottled mineral and spring waters for long periods of storage at 4, 10, and 23 degrees C. The ability of EAEC to survive in bottled water indicates that the source water for bottling industries must be kept free of contamination. Furthermore, the refrigeration of bottled water is recommended to minimize the growth of EAEC in water. PMID- 12636309 TI - Effects of chilling rate on outgrowth of Clostridium perfringens spores in vacuum packaged cooked beef and pork. AB - Cooked, chilled beef and cooked, chilled pork were inoculated with three strains of Clostridium perfringens (NCTC 8238 [Hobbs serotype 2], NCTC 8239 [Hobbs serotype 3], and NCTC 10240). Inoculated products were heated to 75 degrees C, held for 10 min in a circulating water bath to heat activate the spores, and then chilled by circulating chilled brine through the water bath. Samples were chilled from 54.4 to 26.6 degrees C in 2 h and from 26.6 to 4.4 degrees C in 5 h. Differences in initial C. perfringens log counts and log counts after chilling were determined and compared with the U.S. Department of Agriculture (USDA) stabilization guidelines requiring that the chilling process allow no more than 1 log total growth of C. perfringens in the finished product. This chilling method resulted in average C. perfringens increases of 0.52 and 0.68 log units in cooked beef and cooked pork, respectively. These log increases were well within the maximum 1-log increase permitted by the USDA, thus meeting the USDA compliance guidelines for the cooling of heat-treated meat and poultry products. PMID- 12636310 TI - Aspergillus carbonarius as the main source of ochratoxin A contamination in dried vine fruits from the Spanish market. AB - Ochratoxin A (OTA) can occur in a wide range of foods, but unexpectedly high concentrations have been detected in dried vine fruits of various origins. The European Union has recently established a maximum OTA limit of 10 microg/kg for these foodstuffs. In order to determine the likely origin of OTA, a mycological study of 50 dried fruit samples (currants, raisins, and sultanas) representative of the Spanish market was conducted. Fungal contamination was detected in 49 of 50 (98%) samples. Black aspergilli were isolated from all of the positive samples. Aspergillus niger var. niger was isolated from 98% of the samples, and Aspergillus carbonarius was found in 58% of the samples. One hundred sixty-eight A. niger var. niger isolates and 91 A. carbonarius isolates were screened for their ability to produce OTA. Eighty-eight (96.7%) A. carbonarius isolates and one (0.6%) A. niger var. niger isolate were found to be OTA producers. Black aspergilli were the dominant fungi. Among black aspergilli, A. carbonarius has shown a consistent ability to produce OTA and is the most probable source of this mycotoxin in these substrates. PMID- 12636311 TI - Hepatitis A virus detection in oysters (Crassostrea gigas) in Santa Catarina State, Brazil, by reverse transcription-polymerase chain reaction. AB - Shellfish are readily contaminated with viruses present in water containing sewage because of the concentration effect of filter feeding. Hepatitis A virus (HAV) is the main cause of acute hepatitis worldwide and may lead to severe illness or even death. It is transmitted through fecal and oral routes and causes widespread endemic and asymptomatic infections in young children. Here we describe a method for the detection of HAV RNA in shellfish involving the extraction of total RNA from oyster meat followed by reverse transcription polymerase chain reaction (RT-PCR). Virus recovery from oyster extracts artificially seeded with HAV strain HM 175 was examined by RT-PCR. The minimum detection limit was 3.3 focus-forming units of HAV, and the recovery rate was 75.7%. This method was used to assess the viral contamination of four shellfish beds in Santa Catarina State, Brazil, over a 1-year period. Six (22%) of 27 samples collected in autumn and winter from one shellfish bed tested positive for HAV. PMID- 12636312 TI - Rapid detection of Escherichia coli O157:H7 inoculated in ground beef, chicken carcass, and lettuce samples with an immunomagnetic chemiluminescence fiber-optic biosensor. AB - A biosensor was evaluated with regard to its usefulness in the rapid detection of Escherichia coli O157:H7 inoculated in ground beef, chicken carcass, and romaine lettuce samples. The biosensor consisted of a chemiluminescence reaction cell, a fiber-optic light guide, and a luminometer linked to a personal computer in conjunction with immunomagnetic separation. The samples inoculated with E. coli O157:H7 were first centrifuged and suspended in buffered peptone water and then incubated with anti-E. coli O157 antibody-coated magnetic beads and horseradish peroxidase (HRP)-labeled anti-E. coli O157 antibodies to form antibody-coated bead-bacterium-HRP-labeled antibody sandwich complexes. Finally, the sandwich complexes were separated from the samples in a magnetic field and reacted with luminol in the reaction cell. The number of E. coli O157:H7 cells was determined by collecting the HRP-catalyzed chemiluminescence signal from the bead surface through a fiber-optic light guide and measuring the signal with a luminometer. The chemiluminescence biosensor was specific for E. coli O157:H7 in samples containing other bacteria, including Salmonella Typhimurium, Campylobacter jejuni, and Listeria monocytogenes. The chemiluminescence signal was linear on a log scale from 10(2) to 10(5) CFU of E. coli O157:H7 per ml in samples. Detection could be completed within 1.5 h without any enrichment. The detection limits for ground beef, chicken carcass, and lettuce samples were 3.2 x 10(2), 4.4 x 10(2), and 5.5 x 10(2) CFU of E. coli O157:H7 per ml, respectively. PMID- 12636314 TI - Side by side. PMID- 12636313 TI - Evaluation of the 1-2 test for detecting Salmonella in swine feces. AB - The 1-2 Test is a rapid culture test for the detection of motile Salmonella. The aim of this study was to evaluate the 1-2 Test for its ability to detect Salmonella in swine feces following preenrichment and selective enrichment. Pooled pen fecal samples (n = 118) and pig rectal swabs (n = 51) were cultured for Salmonella by the 1-2 Test, which was compared with the standard isolation protocol currently used in our laboratory. In addition, pen fecal samples known to be free of Salmonella were spiked with various concentrations of Salmonella enterica subsp. enterica serovar Typhimurium and cultured by both methods to determine the minimum number of organisms needed to produce a positive result. When naturally contaminated pen feces and rectal swabs were used, results obtained with the standard culture method were similar to those obtained with the 1-2 Test. However, the 1-2 Test did outperform the standard culture method when the spiked feces samples were tested (chi2 = 4.00). The test kit reduced the time and materials required for the detection of Salmonella in swine feces. The results of this study indicate that the 1-2 Test is an accurate method for monitoring Salmonella in swine feces. PMID- 12636315 TI - California children and oral health: trends and challenges. AB - This paper highlights fundamental elements of the challenge of securing optimal oral health for California's children, now and in the future, and underscores the need for new strategies to reduce the risk of dental disease and expand access to effective services. Developing effective strategies to gain optimal oral health for California children, especially those that are most vulnerable to dental disease and its consequences, requires an appreciation of key dynamic factors that define this challenge and the degree to which these factors are modifiable in the near and long term. Particular attention is directed toward three principal elements: population demographics, levels of dental disease in California children, and factors affecting access to services for vulnerable groups of children. The concluding section addresses the need for strategic action. PMID- 12636316 TI - The caries balance: contributing factors and early detection. AB - The aim of this article is to present a brief overview of the dental caries process, in particular, the management of dental caries and the role of early detection methods in the clinical management of caries. PMID- 12636317 TI - Acquisition and transmission of mutans streptococci. AB - Dental caries is an infectious and transmissible disease. The mutans streptococci and some Lactobacillus species are the two groups of infectious agents most strongly associated with dental caries. Earlier studies demonstrated that infants acquire mutans streptococci from their mothers and only after the eruption of primary teeth. More recent studies indicate that mutans streptococci can colonize the mouths of predentate infants and that horizontal, as well as vertical, transmission does occur. These findings will likely facilitate the development of clinical strategies to prevent or delay infant infection by these organisms, thereby reducing the prevalence of dental caries. PMID- 12636318 TI - Early childhood caries: an overview with reference to our experience in California. AB - Dental caries in young children frequently appears first on the maxillary primary incisors, where the liquids ingested by the infant sucking on a bottle or breast remain pooled away from salivary flow. This so-called early childhood caries can rapidly progress to result in rampant destruction of the primary dentition. ECC can affect children from all socioeconomic classes but is most often found in children of new immigrants or those with lower socioeconomic status. The treatment of ECC is costly, and treatment relapses are frequent. Prevention and treatment of ECC focus on inhibition of the growth of oral bacteria. Strategies include reducing the frequency of exposure to bacterial substrates, as well as controlling the growth of oral bacteria. Topical antibacterials such as 10 percent povidone iodine show promise for future use in the inhibition of ECC in young children. PMID- 12636319 TI - The role of caries prevention protocol in pediatric dentistry specialty programs. AB - Advanced specialty education programs in pediatric dentistry are often overwhelmed with patients who need restorative and surgical care, often on an emergency basis. Still, the Commission on Dental Accreditation Standards for Advanced Specialty Education Programs requires that residents receive didactic and clinical training in the prevention of dental caries. This paper contains several recommendations for strengthening the training of pediatric dental residents in caries risk assessment and prevention, including the suggestion that pediatric dental training programs become "dental homes" for their patients. PMID- 12636320 TI - Dental disease prevention and people with special needs. AB - People with special needs are the most underserved of the underserved in our society. They have more dental disease, more missing teeth, and more difficulty obtaining dental care than other segments of the population. Many individuals and groups, including the authors of this paper, have developed community-based systems to improve oral health for people with special needs. However, these systems have not been as successful as they might be because of lack of effective preventive protocols specifically designed for people with special needs. This paper reviews strategies for overcoming informational, physical, and behavioral barriers to oral health and presents a summary of the results of a conference titled "Practical Preventive Protocols for Prevention of Dental Disease in People with Special Needs in Community Settings." The rationale for using an Oral Health Care Plan is presented as well as a sample plan. These strategies and protocols are designed to complement the system of supported community-based oral health care. The goal of this system is to help people with special needs enjoy a lifetime of oral health the same as other members of our society. PMID- 12636321 TI - Don't make me take off my belt. PMID- 12636322 TI - Filling of the bladder for pelvic sonograms: an ancient form of torture. PMID- 12636323 TI - Transvaginal gray scale and color Doppler sonography in primary ovarian cancer and metastatic tumors to the ovary. AB - OBJECTIVE: To compare gray scale and color Doppler features of primary and metastatic ovarian carcinomas. METHODS: Clinical, sonographic (gray scale and color Doppler), and histopathologic data of 143 patients with primary (n = 127 adnexal masses) and metastatic (n = 34 adnexal masses) ovarian cancer were reviewed. Morphologic gray scale parameters assessed were bilaterality, tumor volume, echogenicity, and presence of septa, papillary projections, or solid areas. Color Doppler parameters were presence of blood flow, tumor blood flow location (central versus peripheral), subjective impression of blood flow amount (scanty, moderate, or abundant), lowest resistive index, lowest pulsatility index, and maximal peak systolic velocity (centimeters per second). RESULTS: No statistical differences were found in bilaterality, tumor volume, presence of septa, papillary projections or solid areas, presence of blood flow, tumor blood flow location, subjective impression of blood flow amount, lowest resistive index, lowest pulsatility index, and maximal peak systolic velocity. Metastatic carcinomas were more frequently purely solid tumors (47% versus 26%; P = .001; likelihood ratio, 2.4; 95% confidence interval, 1.2-4.7). CONCLUSIONS: The presence of a purely solid tumor indicates a higher probability of metastatic carcinoma than primary ovarian cancer. However, with the use of gray scale and color Doppler sonography, it is difficult to differentiate primary ovarian carcinomas from metastatic tumors to the ovary. PMID- 12636324 TI - Three-dimensional sonographic morphologic assessment in complex adnexal masses: preliminary experience. AB - OBJECTIVE: To evaluate the role of three-dimensional transvaginal sonography in assessing complex adnexal masses. METHODS: Forty-one women (mean age, 49.5 years; range, 23-75 years) with the diagnosis of complex adnexal masses on the basis of two-dimensional transvaginal sonography were reevaluated by three-dimensional transvaginal sonography. Two different sonologists evaluated the two- and three dimensional transvaginal sonograms. Criteria indicative of malignancy included the presence of gross papillary projections, solid areas, and solid echogenicity for both techniques. Three women (7%) had bilateral masses, giving a total of 44 masses that were ultimately assessed. A definitive histologic diagnosis was obtained in every case after surgical tumor removal. RESULTS: Twenty-one tumors (47.7%) were proved malignant, and 23 (52.3%) were benign. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy for two- and three-dimensional transvaginal sonography were 90%, 61%, 68%, 87%, and 75% and 100%, 78%, 81%, 100%, and 89%, respectively. There were no statistical differences between two- and three-dimensional transvaginal sonography (McNemar test, P = .687). The agreement between both examiners was high (kappa index = 0.71). CONCLUSIONS: The use of three-dimensional transvaginal sonography does not significantly improve the two-dimensional transvaginal sonographic morphologic assessment of complex adnexal masses; however, we found it useful for reinforcing initial diagnostic impressions. PMID- 12636325 TI - Antenatal detection of skeletal dysplasias. AB - OBJECTIVE: To assess the accuracy of the prenatal diagnosis of skeletal dysplasias. METHODS: All antenatally detected anomalies are coded in our ultrasound database, which is linked with a genetics database that includes outcomes. A final diagnosis is sought on the basis of radiographic studies, molecular testing, or both. Our ultrasound and genetics databases were queried for "skeletal dysplasias." All cases were reviewed specifically for the degree of bone shortening and other distinguishing characteristics on antenatal sonography. RESULTS: Thirty-seven cases of skeletal dysplasia were antenatally diagnosed over an 8-year period. Complete follow-up was available in 31 cases. The mean gestational age at diagnosis was 22.7 weeks (range, 14-32.3 weeks). Twenty-one cases were diagnosed before 24 weeks. A final diagnosis was obtained in 80% of cases. The antenatal diagnosis was correct in 20 (65%) of 31 cases. There were 2 false-positive diagnoses. Specific final diagnoses included thanatophoric dysplasia (8), osteogenesis imperfecta (6), Roberts syndrome (2), achondroplasia (3), Ellis-van Creveld syndrome (1), metaphyseal dysplasia (1), spondyloepiphyseal dysplasia (1), distal arthrogryposis (1), caudal regression (1), and glycogen storage disorder (1). The condition was correctly thought to be lethal in 16 of the fetuses on the basis of early severe long bone shortening (13), femur length-abdominal circumference ratio of less than 0.16 (12), hypoplastic thorax (10), marked bowing or fractures (4), short ribs (4), caudal regression (1), and cloverleaf skull (1). The ability to predict lethality was 100%. There were no false-positive findings with respect to lethality. CONCLUSIONS: Accurate antenatal diagnosis of skeletal dysplasias is problematic; in this series, only 20 of 31 cases were correctly diagnosed. However, the antenatal prediction of lethality was highly accurate. The most common predictors of lethal skeletal dysplasias included early and severe shortening of the long bones, femur length-abdominal circumference ratio of less than 0.16, hypoplastic thorax, and certain distinguishing characteristics. PMID- 12636326 TI - Prospective evaluation of the value of combined mammographic and sonographic assessment in patients with palpable abnormalities of the breast. AB - OBJECTIVE: To evaluate the role of combined mammographic and sonographic imaging in patients with palpable abnormalities of the breast. METHODS: Four hundred eleven consecutive cases of palpable abnormalities of the breast underwent combined mammographic and sonographic evaluation. Patients who did not undergo biopsy had imaging and clinical follow-up; the mean follow-up period was 28.9 months (range, 24-33 months). RESULTS: One hundred sixty-five (40.1%) of 411 palpable abnormalities had a benign assessment; 97 (58.7%) of the 165 benign lesions were visible on both mammography and sonography; 66 (40%) of 165 benign lesions were mammographically occult and identified at sonographic evaluation. In 60 (14.6%) of the 411 cases, imaging evaluation resulted in a suspicious assessment; 49 (81.7%) of the 60 lesions categorized as suspicious underwent biopsy; 14 (28.5%) of 49 lesions were histologically proved to be carcinoma. Nineteen (31.6%) of the 60 lesions categorized as suspicious were mammographically occult and identified only on sonography; 14 (73.7%) of these 19 lesions underwent biopsy; 12 (63.1%) of 19 were benign, and 2 (10.5%) were malignant. One hundred eighty-six (45.2%) of the 411 palpable abnormalities had negative imaging assessment findings; 12 patients with negative imaging findings underwent biopsy, and all had benign findings. The sensitivity (14 of 14) and negative predictive value (186 of 186) for a combined mammographic and sonographic assessment were 100%; the specificity was 80.1% (186 of 232). CONCLUSIONS: Cancer was diagnosed in 14 (3.4%) of 411 women who underwent combined imaging for palpable abnormalities of the breast. Combined mammographic and sonographic assessment was shown to be very helpful in identifying benign as well as malignant lesions causing palpable abnormalities of the breast. PMID- 12636328 TI - Modifying the cosine model for nonstationary blood flow signal simulation. AB - OBJECTIVE: The most important features of the Doppler signal are its nonstationary nature and randomness, which make simulating these kinds of signals difficult. Our intent was to create realistic simulations of these signals. METHODS: We generated the Doppler signal for the carotid artery by using the cosine model. We also modified the cosine model to simulate the femoral Doppler signal for the whole cardiac cycle. RESULTS: In auditory comparisons, the simulation results compared favorably with the original data. No appreciable differences were found between the real clinical signals and the simulations for the normal carotid and femoral arteries. However, the calculated normalized root mean square errors were high. CONCLUSIONS: The cosine model is simple to implement and requires input of only the spectrum shape variation during a cardiac cycle. In addition, it is more informative than other modeling methods and therefore is preferred over other methods. PMID- 12636327 TI - Echocardiography in early pregnancy: review of literature. AB - OBJECTIVE: First-trimester transvaginal sonography is a widely used technique to examine the fetus early in pregnancy. The aim of this review was to explore the possibilities of examining the fetal heart at this early stage of pregnancy. METHODS: With the use of a computerized database (PubMed, US National Library of Medicine, Bethesda, MD), articles on first-trimester echocardiography were retrieved. Furthermore, the cited references of the studied articles were used to find additional articles. RESULTS: In the analyzed articles, an increase in visualization rates of the 4-chamber view and the outflow tracts was shown in the last decade, with visualization rates of greater than 90% at 13 weeks' gestation. The different cardiac defects that are shown in first-trimester fetuses are mainly defects involving the 4-chamber view, indicating that defects solely affecting the outflow tracts are difficult to diagnose in the first trimester of pregnancy. The sonographic assessment of the fetal heart between 14 and 18 weeks' gestation has been described. The literature, however, has not shown clear advantages of performing fetal echocardiography during this period compared with transvaginal sonography at 13 weeks' gestation. CONCLUSIONS: First-trimester echocardiography is a promising technique, which can be of considerable value for patients at risk of having offspring with cardiac defects. This technique is, however, currently limited to a few specialized centers. PMID- 12636329 TI - Diagnosis of pregnancy-associated uterine venous plexus thrombosis on the basis of transvaginal sonography. AB - OBJECTIVE: To describe the sonographic signs of uterine venous plexus thrombosis. METHODS: Four pregnant patients had a diagnosis of uterine venous plexus thrombosis in the first half of gestation. The diagnosis was based on transvaginal sonography only in 3 cases, and the fourth had magnetic resonance imaging corroboration. RESULTS: All 4 patients had similar sonographic features of uterine venous plexus thrombosis on transvaginal sonographic examination. The thrombi within the dilated veins were shown as elongated echogenic structures along the lumen that appeared round on transverse views of the affected veins. They showed swinging movements provoked by gentle transducer pressure. Power and color Doppler sonography enhanced the uterine venous plexus thrombosis diagnosis by showing blood flow around the thrombi. There were no signs of thromboembolic disease. Sonographic findings in deep leg veins and iliac veins were normal in all cases. Complete thrombophilia studies did not reveal any abnormalities. The uterine venous plexus thrombosis could not be detected on transabdominal sonography and was shown better by transvaginal sonography compared with magnetic resonance imaging. During 3 months of anticoagulation therapy, the thrombi gradually disappeared in all cases. CONCLUSIONS: Focusing on the pelvic veins while performing a transvaginal sonographic study during pregnancy may reveal important findings, which may have clinical implications. The therapeutic treatment of uterine venous plexus thrombosis is controversial and still empirical. PMID- 12636331 TI - Benign testicular lobulation: sonographic findings. PMID- 12636330 TI - Intratesticular arteriovenous malformation: color Doppler sonographic findings. PMID- 12636332 TI - Adenomatoid tumor of the tunica vaginalis testis: a special maneuver in diagnosis by ultrasonography. PMID- 12636333 TI - Mammographic and sonographic findings of a breast cystic lymphangioma. PMID- 12636334 TI - First-trimester diagnosis of osteogenesis imperfecta type II by three-dimensional sonography. PMID- 12636335 TI - Superior sternal cleft associated with PHACES syndrome: postnatal sonographic findings. PMID- 12636336 TI - A pox upon human folly. PMID- 12636337 TI - Pre-event smallpox vaccination: unresolved issues. PMID- 12636338 TI - Vertebral compression fractures what time destroys, methylmethacrylate may mend. PMID- 12636339 TI - What are the key issues women face when ending hormone replacement therapy? PMID- 12636340 TI - What is the appropriate treatment for Dupuytren contracture? PMID- 12636341 TI - How do you manage a healthy, asymptomatic, 24-year-old with a positive RPR on a premarital blood test? PMID- 12636342 TI - Worsening shortness of breath. PMID- 12636343 TI - Fungal nail infection: assessing the new treatment options. AB - Onychomycosis can be improved or eradicated with appropriate treatment. Newer oral antifungal drugs are highly effective and have few adverse effects, although care in prescribing is needed because of potential drug interactions and hepatobiliary dysfunction. PMID- 12636344 TI - Challenges and choices in drug therapy for chronic pain. AB - By treating chronic pain effectively, physicians can improve the quality of their patients' lives considerably. This article reviews the mechanisms and treatment of chronic pain, with emphasis on overcoming the barriers to effective analgesia. PMID- 12636345 TI - Improving care of chronic heart failure: advances from drugs to devices. AB - The right combination of drugs and surgical treatment can improve systolic function and prevent, attenuate, or reverse heart failure. Patient education and disease management programs can reduce hospitalizations. Optimal treatment for each patient is guided by a thorough evaluation and use of functional classification and disease staging systems. PMID- 12636346 TI - Vertebral compression fractures: manage aggressively to prevent sequelae. AB - New drugs to treat osteoporosis, along with two new minimally invasive surgical procedures, are important options for preventing vertebral compression fractures and treating severe back pain and disability. However, the mainstay treatments remain cautious use of analgesics, limited bed rest, and physical rehabilitation. PMID- 12636347 TI - The effects of host immunity on virulence-transmissibility relationships in the rodent malaria parasite Plasmodium chabaudi. AB - Here we examined the impact of host immunity on relationships between parasite virulence, transmission rate, intrinsic growth rate and host recovery rate in the rodent malaria parasite, Plasmodium chabaudi. Groups of naive and immunized mice were infected with 1 of 10 cloned lines of parasites and their infection dynamics were monitored for 19 days. We found that (1) host immunity reduced the growth rate, virulence, transmission rate and infection length, with a consequent 3-fold reduction in life-time transmission potential, (2) clone means for these traits ranked similarly across naive and immunized mice, (3) regression slopes of transmission potential on growth rate, virulence and infection length were similar in naive and immunized mice, (4) virulence and infection length were positively correlated in immunized but not naive mice, and (5) for a similar level of parasite growth rate and virulence, transmission potential and infection length were lower in immunized than naive mice. Thus host immunity reduced all these fitness traits in a manner consistent with direct parasite-driven biological links among them. These results support the basic assumption underlying our theory that predicts that anti-disease vaccines will select for higher virulence in those microparasites for which virulence is integrally linked to transmission. PMID- 12636348 TI - The molecular evidence of Babesia microti infection in small mammals collected in Slovenia. AB - In Europe, the zoonotic cycle of Babesia microti has not been determined so far. Recently, B. microti was detected in Ixodes ricinus ticks in Slovenia by using molecular methods. In order to investigate the mammalian hosts of B. microti in Slovenia we collected 261 small mammals representing 11 species. They were tested for the presence of babesial parasites with a PCR assay based on the nuclear small subunit rRNA gene (nss-rDNA). The bank vole (Clethrionomys glareolus) and yellow-necked mouse (Apodemus flavicollis) were infected with B. microti. The prevalence rate was 15.9% for C. glareolus and 11.8% for A. flavicollis. Nucleotide sequences of amplified portions of B. microti nss-rDNA from C. glareolus and A. flavicollis were indistinguishable from each other and identical with those previously described in I. ricinus ticks collected in Slovenia. The results of this study represent molecular evidence of B. microti in small mammals in Europe. PMID- 12636349 TI - The impact of a vertically transmitted microsporidian, Nosema granulosis on the fitness of its Gammarus duebeni host under stressful environmental conditions. AB - Although purely vertically transmitted parasites are predicted to cause low pathogenicity in their hosts, the effects of such parasites on host fitness under stressful environmental conditions have not previously been assessed. Here, we investigate the effects of Nosema granulosis, a vertically transmitted, microsporidian parasite of the brackish water amphipod Gammarus duebeni, on host growth and survival under conditions of host-host competition and limited food. The parasite had no effect on host survival, but caused a reduction in juvenile growth. Stressful environmental conditions also led to a reduction in G. duebeni growth. However, we found no evidence to support the prediction that parasitized hosts would suffer a greater reduction in fitness than uninfected hosts under adverse environmental conditions. We interpret our results in the context of selection for successful vertical parasite transmission. PMID- 12636350 TI - Philasterides dicentrarchi (Ciliophora:Scuticociliatida) expresses surface immobilization antigens that probably induce protective immune responses in turbot. AB - Philasterides dicentrarchi is a histophagous ciliate causing systemic scuticociliatosis in cultured turbot. This study demonstrates that turbot which survive this disease have serum antibodies that recognize ciliary antigens of this ciliate in ELISA and immobilize/agglutinate the ciliate in vitro. Mouse sera raised against ciliary antigens and integral membrane proteins are likewise capable of immobilizing/agglutinating the ciliates, indicating that P. dicentrarchi, like other ciliates, expresses surface immobilization antigens. Furthermore, the antigen agglutinating reaction induces the parasite to shed its surface antigens rapidly, replacing them with others with different specific serology. This antigen shedding and variation response is similar to that detected in other protozoan parasites. Immunization of turbot with ciliate lysate plus adjuvant or with formalin-fixed ciliates induced synthesis of agglutinating antibodies and conferred a degree of protection against challenge infection, suggesting that the response to surface antigens may play an important role in defence against this pathogen, SDS-PAGE and immunoblotting studies indicated the existence of a predominant polypeptide of about 38 kDa in the ciliary antigen and membrane protein fractions, and this may be the principal surface antigen of P. dicentrarchi. PMID- 12636351 TI - Delayed tail loss during the invasion of human skin by schistosome cercariae. AB - Schistosomiasis is initiated when cercarial larvae invade human skin. Contrary to long-held assumptions, most cercariae of Schistosoma mansoni do not shed their propulsive tails as they penetrate. Scanning electron microscopy studies and infection experiments with entire human skin and differentiated, stratum corneum like, human keratinocyte cultures, have shown that most cercarial tails enter the skin along with their bodies. We propose that this behaviour is an adaptive trait linked with concomitant immunity. PMID- 12636352 TI - Fasciola hepatica: surface and internal tegumental changes induced by treatment in vitro with the sulphoxide metabolite of albendazole ('Valbazen'). AB - A morphological study has been carried out to determine the effect of the active sulphoxide metabolite of the benzimidazole anthelmintic, albendazole (ABZ-SO) on the adult liver fluke, Fasciola hepatica. Whole flukes were treated with ABZ-SO for 12 and 24 h at a concentration of 10 microg/ml. The changes in response to drug treatment were examined by scanning electron microscopy (SENI), transmission electron microscopy (TEM) and tubulin immunocytochemistry (ICC). No surface changes were apparent following 12 h ABZ-SO treatment, but localized blebbing was observed after 24 h, which became more extensive towards the posterior region of both surfaces. TEM of sections from the posterior midbody region revealed that ABZ-SO caused the accumulation of secretory bodies in the tegumental cells and in their cytoplasmic connections and, after 24 h, just above the basal plasma membrane. Localized blebbing of the apical membrane also occurred. The morphology of the Golgi complexes within the tegumental cells began to change after 12 h treatment with ABZ-SO and, by 24 h, few complexes were observed. A distinct increase in tubulin immunoreactivity occurred after 12 h treatment, but this decreased after 24 h. The results obtained are consistent with those expected for microtubule inhibition. They are discussed in relation to the action of established microtubule inhibitors, as well as the benzimidazole derivative, triclabendazole. PMID- 12636353 TI - A Boophilus microplus vitellin-degrading cysteine endopeptidase. AB - Here we describe the purification and characterization of a vitellin (VT) degrading cysteine endopeptidase (VTDCE) from eggs of the hard tick Boophilus microplus. A homogeneous enzyme preparation was obtained by chromatographic fractionation on ion-exchange and gel filtration columns and an autolysis step. This step consisted of incubation of a semipurified enzyme (after the first ion exchange chromatography) at pH 4.0 that dissociated the enzyme from VT, to which VTDCE is naturally tightly associated. The enzyme purity was confirmed by capillary and native gel electrophoresis, and SDS-PAGE suggested the enzyme is a dimer of 17 and 22 kDa. VTDCE was active upon several synthetic substrates, with a preference for a hydrophobic or a basic residue in P1, and a hydrophobic residue in P2. VTDCE also hydrolysed haemoglobin, albumin, gelatin and vitellin. VTDCE is inactive in the absence of DTT and was totally inhibited by E-64, indicating it is a cysteine endopeptidase. Our results suggest that VTDCE is a major enzyme involved in yolk processing during B. microplus embryogenesis. PMID- 12636354 TI - Secreted adhesion molecules of Strongyloides venezuelensis are produced by oesophageal glands and are components of the wall of tunnels constructed by adult worms in the host intestinal mucosa. AB - The parasitic female of Strongyloides venezuelensis keeps invading the epithelial layer of the host intestinal mucosa. Upon invasion, it adheres to the surface of the intestinal epithelial cells with adhesion molecules secreted from the mouth. It has been demonstrated that S. venezuelensis are expelled from the intestine because mucosal mast cells inhibit the attachment of adult worms to the mucosal surface. In the present study, we generated specific antibodies against secreted adhesion molecules to investigate their function in vivo, because these molecules have been demonstrated only in vitro in spite of the importance in the infection processes. A mouse monoclonal antibody specific to S. venezuelensis adhesion molecules inhibited the attachment of adult worms to plastic dishes and the binding of adhesion molecules to rat intestinal epithelial cells. Immunohistochemical study revealed that adhesion molecules were produced by oesophageal glands and were continuously secreted in vivo to line the wall of the tunnels formed by adult worms in the intestinal mucosa. Our findings indicate that adhesion molecules play essential roles in the infection processes of S. venezuelensis in the host intestine. PMID- 12636355 TI - Is body size or activity of copepods related to ingestion of parasite larvae? AB - This study shows that ingestion of Schistocephalus solidus coracidia was related to general activity of Macrocyclops albidus copepods at the time of exposure. The lower the activity of the host, the fewer parasites it ingested. In an earlier study it was shown that large M. albidus copepods were less likely to become infected with S. solidus than small copepods, which could potentially be caused by differential ingestion of parasites. However, the current study did not show any evidence for such an effect arising through differential ingestion. Body size was not related to ingestion of parasites, but was positively correlated to activity. So, even though size did not significantly relate to ingestion of parasites, if anything, through their higher activity large copepods rather than small copepods may have ingested more parasites. This study indicates that differences in resistance to this parasite do not come about through differential ingestion of parasites. Also, an earlier study failed to show differential elimination of the parasite from the haemocoel. This leaves avoidance of penetration through the gut wall as the most plausible candidate causing large copepods to be more resistant to this parasite than small copepods. PMID- 12636356 TI - Hookworm cathepsin D aspartic proteases: contributing roles in the host-specific degradation of serum proteins and skin macromolecules. AB - Cathepsin D aspartic proteases of hookworms were recently implicated in the host specific digestion of haemoglobin by adult parasites. Ac-APR-1 from the dog hookworm, Ancylostoma caninum and Na-APR-1 from the human hookworm, Necator americanus, were shown to be expressed in the infective larval stage (L3) as well as adult worms. We now show that both proteases degraded skin macromolecules and serum proteins, some of which were cleaved more readily from permissive definitive hosts as opposed to non-permissive hosts. Na-APR-1 degraded human collagens more efficiently than did Ac-APR-1, and Ac-APR-1 degraded canine serum albumin more efficiently than did Na-APR-1. On the other hand, both enzymes degraded human serum proteins (albumin and fibrinogen) with approximately equal efficiency under the conditions of our assays in vitro. Molecular models of these 2 orthologous, aspartic proteases showed that, despite having active site clefts with identical primary sequences, residues in the S3 pocket adopted different conformations, likely accounting for different substrate preferences reported previously. Antisera raised to both proteases partially inhibited (16-26%) migration of hookworm L3 through hamster skin in vitro, further implying a connective tissue invasive role for these enzymes in addition to digestion of serum and erythrocyte proteins for nutrition. PMID- 12636357 TI - The effect of treatment with a 1% injectable formulation of moxidectin during the rainy season and at the beginning of the dry season on gastrointestinal nematodes in cattle from communal areas in Zimbabwe. AB - The effect of 1% moxidectin/cydectin at 0.2 mg/kg live weight on gastrointestinal nematodes and on the growth of calves, weaners and cows was investigated in five communal areas on the highveld of Zimbabwe. Three field experiments were carried out between March 1996 and June 1997. In experiment 1, treatment was administered in all five areas at the end of the rainy season in March 1996, followed by a further treatment at the beginning of the dry season in May/June 1996. In experiment 2, the treatment was administered in three areas at the end of the rainy season in March 1997. In experiment 3, treatment was administcred in one area at the beginning of the dry season in April 1997. Large numbers of eggs were present in the faeces of calves and weaners at the start of experiments 1 and 2. Epg values were lower in cows and in all age categories in experiment 3. There was a statistically significant reduction in epg values in calves, weaners and cows following treatment with a reduction of 90-99% in all cases except in cows in experiment 3, where no meaningful assessment was possible owing to the low egg counts in both the treated and control cows. The dominating larval types in faecal cultures were Cooperia and Haemonchus. Trichostrongylus, Oesophagostomum and Bunostomum were also found. Following treatment, Haemonchus was suppressed far more than Cooperia. This may be related to a longer residual effect against abomasal parasites like Haemonchus in comparison to small intestinal worms like Cooperia. Anthelmintic treatment conferred significant weight gain advantages (p < 0.05) on treated calves. weaners and cows. The weight gains are discussed in relation to disease and nutrition. PMID- 12636358 TI - Evolution of high-level, multiple anthelmintic resistance on a sheep farm in Malaysia. AB - Anthelmintic resistance in nematode parasites of sheep and goats on a government farm in north Malaysia was monitored over a 3-year period (1997-2000). The faecal egg count reduction test (FECRT) was conducted on young sheep at the beginning and end of this period. Changes in management, designed to reduce the selection pressure for the development of anthelmintic resistance, were also implemented during this time. By far the most important parasite problem was Haemonchus contortus. In 1997, this nematode was found to be resistant to levamisole, with suspected resistance to closantel and moxidectin. However, when the FECRT was repeated 3 years later, its resistance status had become much more severe, with resistance to benzimidazole, levamisole and ivermectin, and suspected resistance to moxidectin. This rapid evolution to multiple anthelmintic resistance is a major concern that needs to be arrested. There is an urgent need to evaluate other control strategies that incorporate livestock management, the 'smart' use of drugs and non-chemotherapeutic approaches, such as biological control agents. PMID- 12636359 TI - Biodiversity of domestic livestock in the Republic of Yemen. AB - This paper describes the domestic livestock of the Republic of Yemen and aspires to complement earlier sources listing or partially describing 'breeds'. It attempts to cover all species and provide indications of production parameters through a literature review and via field observations made by the author in 1999. Information is provided on livestock numbers and the economic importance of animal production. Most animals are kept in sedentary mixed crop-livestock production systems; transhumant systems have the next greatest number of stock; with nomadic systems being of least and declining importance. Yemen's livestock appear to comprise at least 11 breeds of sheep, 5 breeds of goat, 2 breeds of cattle, 4 breeds of camel, 2 breeds of donkey and 1 breed of horse. There are no data on breeds of poultry but domestic fowl (where clearly considerable diversity exists) and pigeons are kept. There is little formal information on the history and relationships of most breeds. Some appear to be of ancient local origin, whereas others show affinities with those of neighbouring and other countries. None of the identified types is considered endangered, so conservation would be premature. A more formal and detailed genetic characterization, to add to the largely morphological and traditional classification, may, however, reveal such a need. PMID- 12636360 TI - Animal health and disease control in the Usangu Wetland of Southwestern Tanzania. AB - The Usangu Wetland in the Southern Highlands of Tanzania has always been a major livestock production area. This paper describes the physical and social enviroment of these Plains before presenting a short history of the veterinary services in the area. The main part of the paper examines, through historical records and interviews with livestock owners and administrative officials, the history of the major diseases affecting livestock. PMID- 12636361 TI - Effect of restricted suckling of calves on the productivity of crossbred dairy cattle. AB - An experiment on restricted suckling of crossbred dairy cows was conducted at the Livestock Research Centre, Tanga in north-east Tanzania. The objective of the experiment was to evaluate the comparative productivity of Bos taurus x Bos indicus cows of medium and high levels of Bos taurus inheritance, whose calves were either bucket-reared or suckled residual milk. Lactation milk yield, length and persistency were 1563 L, 289 days, and 1.0, respectively, for the bucket reared and 1592 L, 289 days and 1.4. respectively, for the suckling group. Days to observed ocstrus, first insemination and conception for cows whose calves were bucket-reared were 47, 74 and 115 days, respectively, and 57, 81 and 126 days, respectively, for the suckling cows. The calf weights were similar at 1 year of age. The productivity of the cows, measured as the annual milk offtake, was not significantly higher for those that suckled their calves than for those whose calves were bucket-reared. PMID- 12636362 TI - The control of parasitic nematodes in commercial piggeries in Kenya as reflected by a questionnaire survey on management practices. PMID- 12636364 TI - Computer-aided design of chiral ligands. Part 2. Functionality mapping as a method to identify stereocontrol elements for asymmetric reactions. AB - A computational method to determine the energetically favorable positions of functional groups with respect to the transition states of stereoselective reactions based on force field energy minimization is presented. The parameters of this functionality mapping, the characteristics of the target transition states, and the features of the probe structures are outlined. Our method was found to reproduce the positions of the stereodiscriminating fragments for some known chiral ligands including the Masamune dimethylborolane, dimenthylborane, the Corey stien reagent, the Roush allylboronate tartrates, and the secondary amine Diels-Alder catalysts described by MacMillan. Functionality mapping can be used to better understand the specific interactions in the transition states leading to the products by providing a quantitative measure of the stabilization/destabilization afforded by the different ligand components via nonbonded interactions. The method can determine if a chiral ligand imparts the observed selectivity by stabilizing one reaction pathway, by destabilizing a reaction pathway, or by a combination of both. Orientational as well as positional information about potential functional groups is readily obtained. In addition to its utility as an analytical tool, functionality mapping can be used to explore starting points for the design of new chiral ligands. PMID- 12636363 TI - Production performance, serum/yolk cholesterol and immune competence of white leghorn layers as influenced by dietary supplementation with probiotic. AB - An experiment was conducted to measure the influence of a dietary probiotic on the production performance, the concentrations of cholesterol in the serum and yolk and immune competence in White Leghorn layers from 25 to 72 weeks of age. One hundred and twenty commercial White Leghorn layers, aged 24 weeks, with an average of 62% hen-day egg production, were equally and randomly distributed into three groups, with eight replicates of 5 birds in each. The birds were reared in individual laying cages. They were placed on one of three dietary treatments: basal, or basal with probiotic supplementation at a rate of 100 or 200 mg/kg feed. The addition of probiotic significantly increased the egg production, shell weight, shell thickness and serum calcium, and reduced the concentrations of cholesterol in the serum and yolk. However, no differences in these traits was observed between the groups receiving 100 or 200 mg probiotic. Feed conversion, egg weight, serum phosphorus and serum alkaline phosphatase activity were not influenced by supplementation with probiotic. Antibody production in response to the inoculation of sheep red blood cells and the cutaneous basophilic hypersensitivity (CBH) responses to inoculation with phytohaemagglutinin did not differ significantly among the dietary groups at either 24 or 40 weeks of age. The antibody titre was significantly higher in the groups supplemented with probiotic at 64 weeks of age. The addition of 100 mg/kg of probiotic in the diet significantly increased the CBH response at 64 weeks of age. PMID- 12636366 TI - Nonplanar aromatic compounds. 8. Synthesis, crystal structures, and aromaticity investigations of the 1,n-dioxa[n](2,7)pyrenophanes. How does bending affect the cyclic pi-electron delocalization of the pyrene system? AB - A series of 1,n-dioxa[n](2,7)pyrenophanes (n = 7-12) with increasingly nonplanar pyrene moieties was synthesized by a 9-10 step sequence starting from 5 hydroxyisophthalic acid. The crystal structure of each member of this series was determined crystallographically. Several spectroscopic properties were found to vary with the extent of the nonplanarity of the pyrene unit. The way in which the distortion from planarity of the pyrene system influences its pi-electron delocalization was investigated by using two quantitative descriptors of aromaticity based on geometry (HOMA) and magnetism (magnetic susceptibility and NICS). Both methods suggest that the aromaticity of the pyrene moiety is diminished only slightly upon increasing the bend angle theta from 0 degrees to 109.2 degrees. PMID- 12636365 TI - Through-shell alkyllithium additions and borane reductions. AB - The through-shell borane reduction and methyllithium addition to benzaldehyde (1), benzocyclobutenone (2), and benzocyclobutenedione (3) incarcerated inside a hemicarcerand (4) with four tetramethylenedioxy bridges are reported. All guests could be reduced and methylated. Selective monoreduction and monomethylation were observed for 3. In the methyllithium addition to 4[symbol: see text]3, the initially formed lithium alcoholate underwent a Moore rearrangement. The reactivity of the incarcerated guests toward methyllithium increased in the order 1 < 2 << 3 and toward borane in the order 1 << 2 approximately equal 3. Guest reactivity was correlated with the inner-phase location of the reacting carbonyl group in the preferred guest inner-phase orientation. The latter was determined from the X-ray structures of 4[symbol: see text]1, 4[symbol: see text]2, and 4[symbol: see text]3, from molecular mechanical calculations, and from the hemicarcerand-induced upfield shift of the guest proton resonances. In the methyllithium and n-butyllithium addition to 4[symbol: see text]1 and 4[symbol: see text]3 at elevated temperatures, selective cleavage of a host's spanner or tetramethylenedioxy bridge, respectively, was observed. The cleavage of one spanner also took place in the methyllithium addition to the 1-methyl-2 pyrrolidinone hemicarceplex. These scission reactions are initiated by the initially formed lithium alcoholates, which show enhanced basicity and nucleophilicity in the inner phase as compared to the bulk phase. Mechanisms for the host scission reactions are discussed. PMID- 12636367 TI - Synthesis, resolution, and absolute stereochemistry of (-)-blestriarene C. AB - A naturally occurring 1,1'-biphenanthrene, blestriarene C (1), was prepared in 13 steps and 30% overall yield. The key steps are the ester-mediated nucleophilic aromatic substitution on 2,6-di-tert-butyl-4-methoxyphenyl 5-isopropoxy-2 methoxybenzoate (4) by 2-methoxy-4-methoxymethoxy-6-methylphenylmagnesium bromide (5) and a novel intramolecular cyclization of the resulting 4-isopropoxy-2' methoxy-4'-methoxymethoxy-6'-methylbiphenyl-2-carboxylic ester 14 to 7-isopropoxy 4-methoxy-2-(methoxymethoxy)phenanthren-9-ol (15). The racemic blestriarene C was optically resolved by chiral HPLC on a preparative scale to give several 10-mg yields of both the enantiomers in up to 95% ee. The absolute stereochemistry was determined to be S(a)-(-) by the axial chirality recognition method, which was based on the stereospecific formation of a 12-membered cyclic diester containing two biaryl-o,o'-diyl unites joined by ester -CO(2)- linkages. The validity of the method was confirmed by an X-ray crystallographic analysis and ab initio conformational analyses of such 12-membered cyclic diesters. It was found that blestriarene C and its 7,7'-diisopropyl ether 2 underwent rapid photoracemization even under ambient light exposure. PMID- 12636368 TI - Bismuth nitrate-catalyzed versatile Michael reactions. AB - Bismuth nitrate-catalyzed versatile Michael reaction was developed to reduce the complications that characterize the current standard Michael reaction and used for facile preparation of organic compounds of widely different structures. For example, several substituted amines, imidazoles, thio compounds, indoles, and carbamates were prepared at room temperature by following this method. In contrast with the existing methods using many acidic catalysts, this method is very general, simple, high-yielding, environmentally friendly, and oxygen and moisture tolerant. However, the promoting role of bismuth nitrate in this reaction is not understood at this time. PMID- 12636369 TI - A short synthetic route to the calystegine alkaloids. AB - An efficient strategy is described for the synthesis of enantiopure calystegine alkaloids. The key step employs a zinc-mediated fragmentation of benzyl-protected methyl 6-iodo-glycosides followed by in situ formation of the benzyl imine and Barbier-type allylation with zinc, magnesium, or indium metal. Stereochemistry in the pivotal allylation is controlled by the choice of the metal. The functionalized 1,8-nonadienes, thus formed, are converted into cycloheptenes by ring-closing olefin metathesis. Regioselective hydroboration and oxidation give the corresponding cycloheptanones, which are deprotected to afford the desired calystegines. Hereby, calystegine B(2), B(3), and B(4) are prepared from D glucose, D-galactose, and D-mannose, respectively. This route constitutes the shortest synthesis of calystegine B(2) and gives rise to the first total syntheses of calystegine B(3) and B(4). PMID- 12636370 TI - Synthesis of the C-linked disaccharide alpha-D-Man-(1-->4)-D-Man employing a SmI(2)-mediated C-glycosylation step: en route to cyclic C-oligosaccharides. AB - Investigations are reported on the assembly of the C-linked disaccharide alpha-D Man-(1-->4)-D-Man, representing the first steps in our projected synthesis of a cyclic C-oligomer containing repeating units of this C-dimer. The key step in this synthesis uses a SmI(2)-mediated coupling of 2,3,4,6-tetra-O-benzyl-alpha-D mannopyranosyl 2'-pyridyl sulfone with a C4-formyl branched mannopyranoside unit, affording the C-disaccharide derivative with complete stereocontrol at the two new stereogenic centers. Subsequently, a modified tin hydride based deoxygenation produced the target carbohydrate analogue. The synthesis of the C4-formyl monosaccharide makes use of a stereoselective radical-based allylation followed by double bond migration and ozonolysis. PMID- 12636371 TI - 2,6-Diaziadamantane: a single-crystal X-ray diffraction study and theoretical calculations. AB - The first single-crystal X-ray diffraction study of 2,6-diaziadamantane (2), a compound containing two diazirine functionalities, and B3LYP/6-311+G* and MP2/6 311+G* calculations relating to the interaction between carbon-nitrogen and nitrogen-nitrogen covalent bonds and electron density within the diazirine rings have been performed. In addition, the protonation of 2-aziadamantane (1) has been investigated. The single-crystal X-ray diffraction study of 2, including UV/vis titrations of 1, corroborate the theoretical conclusion that the low basicity can be attributed to a rehybridization of the nitrogen lone pairs. Moreover, the strain in diazirine is unusually small. PMID- 12636372 TI - A new reactivity-based one-pot synthesis of N-acetyllactosamine oligomers. AB - Poly-N-acetyllactosamine oligomer is a type-2 glycan core from which a number of important bioactive glycoconjugates are assembled in vivo. Development of an effective synthesis of N-acetyllactosamine oligomers will therefore provide a new chemoenzymatic entry to this class of complex saccharides. This paper describes the design and synthesis of thioglycoside building blocks, determination of their relative reactivity values, and demonstration of their use in the programmable one-pot synthesis of various N-acetyllactosamine oligomers. Through a combination of segment condensation, the strategy allows for the preparation of larger oligosaccharides with minimal protecting group manipulation, as illustrated in the synthesis of an octasaccharide in a very short period of time. PMID- 12636373 TI - Multiple component reactions: an efficient nickel-catalyzed Reformatsky-type reaction and its application in the parallel synthesis of beta-amino carbonyl libraries. AB - Multiple-component condensations (MCC) where three or more reactants combine to afford a new core structure possessing the molecular features of its composite building blocks is a powerful method for the preparation of molecular diversity. We have developed an efficient, nickel-catalyzed, Reformatsky-type three component condensation (3CC) reaction that affords beta-amino carbonyl compounds. The scope of the reaction is demonstrated both in the gram and microscale settings; 15 beta-amino esters, amides, and a ketone were prepared efficiently at the mmol scale, and a library of 64 beta-amino carbonyl compounds was generated at the micromol scale. PMID- 12636374 TI - Comparison of the electronic structures of imine and hydrazone side-chain functionalities with the aid of 13C and 15N NMR chemical shifts and PM3 calculations. The influence of C=N-substitution on the sensitivity to aromatic substitution. AB - Benzaldehyde derivatives possessing a C=N double bond in the side-chain of the aromatic ring exhibit a reverse dependence of the (13)C NMR chemical shifts of the C=N carbon on the benzylidenic substituents X. Thus, electron-withdrawing substituents cause shielding (shift is reduced), while electron-donating ones cause deshielding. The origin of this phenomenon, which is in contrast with the idea of the generalized electronic effect, is extensively studied here by comparing the behavior of sets of benzaldehyde derivatives bearing various substitutents Y on the C=N nitrogen (Y-N=CH-C(6)H(4)-X). The effects of substituents X on the C=N unit change when Y is varied. Combination of the influences of the substituents X and Y gives a sensitive balance between the different resonance structures of the compounds. Our graphical treatment, where the rho(I) and rho(R) values observed for substituent X are plotted against the sigma(p)(+) value of substituent Y, is a novel use of Hammett-type substituent parameters. The justification of this method and our conclusions could be verified, for instance, by the fair correlation between the rho(I) or rho(R) values and the atomic charges of the imine carbon of the unsubstitued phenyl derivatives as well as by the correlations of the relevant bond orders and/or bond lengths both with the substituent parameters and with the atomic charges. PMID- 12636375 TI - Preparation of N-aryl azacrown ether derivatives, using arene-iron chemistry. AB - m- or p-phenylenediamine and m- or p-chlorophenyl-substituted azacrown ether derivatives were synthesized through sequential nucleophilic substitution of [(eta(5)-cyclopentadienyl)(eta(6)-(m- or p-dichlorobenzene))]iron hexafluorophosphate by azacrown ethers and cyclohexaamines. Monoarylation is the main reaction for diazacrown ethers. The overall yield from the starting complex is 50-96% for multiple steps. PMID- 12636376 TI - An approach to long and unsubstituted molecular wires: synthesis of redox-active, cationic phenylethynyl oligomers designed for self-assembled monolayers. AB - Various oligo(phenyleneethynylene)s (OPEs) have been synthesized in the past, as they are considered as prototypes of molecular wires. When the oligomers are capped by a redox site at one end and a thiol at the other end, the resulting molecules can be grafted as a self-assembled monolayer on a gold electrode and fully studied by electrochemical techniques. Unfortunately, such molecules are usually poorly soluble and require the incorporation of solubilizing pendant groups. In this paper, we show that the replacement of the classically used redox group ferrocene by a cationic organometallic ruthenium complex, namely, [Ru(bipy)(2)(ppH)](+) (bipy, 2,2'-bipyridine; ppH, 2-(2'-yl-phenyl)pyridine), allows a concise synthesis of an unsubstituted thioacetate-capped OPE up to four repetitive units long. The positive charge does not interfere with the conventional organic chemistry used to elongate, purify, or characterize the hexafluorophosphate salts of the molecules. To our knowledge, this represents the first family of long, poorly substituted OPEs designed for self-assembly. PMID- 12636377 TI - Formation and characterization of a multicomponent equilibrium system derived from cis- and trans-1-aminomethylcyclohexane-1,2-diol. AB - Both cis and trans isomers of amino diols 3-6 were prepared stereoselectively. In the reactions between 3-6 and phenyl isothiocyanate, the ring closure proceeded regioselectively and resulted only in spiro derivatives of 2 phenyliminooxazolidines 9, 10, 13, and 14. The reaction of cis- (or trans-)1 aminomethylcyclohexane-1,2-diol 4 (or 6) with 1 equiv of an aromatic aldehyde 15a g in EtOH at room temperature resulted in a complex, multicomponent equilibrium mixture of 16a-g and 18a-g (or 17a-g and 19a-g), in each case consisting of a five-component, ring-chain tautomeric system 16A-E (or 17A-E), involving the Schiff base, two epimeric spirooxazolidines, two epimeric condensed 1,3-oxazines, and some of the four tricyclic compounds 18A-D (or 19A-D). The five-component, ring-chain equilibria were found to be adequately described by the Hammett-Brown linear free energy equation. PMID- 12636378 TI - New strategy for convergent steroid synthesis. AB - We published recently our results on a new and convergent synthesis of natural steroids. The strategy was based on a cycloaddition reaction of Nazarov reagents 2 and 5 with cyclohexenones 1 and 4. In this paper we report results that deal with the synthesis of two new bicyclic Nazarov reagents (13 and 19) and their cycloaddition with two cyclohexenones (1 and 4). These new results constitute an important improvement concerning the versatility of the strategy since tetracycles having the stereochemistry found in natural steroids are now available. PMID- 12636380 TI - Functionalized arylzinc compounds in ethereal solvent: direct synthesis from aryl iodides and zinc powder and application to Pd-catalyzed reaction with allylic halides. AB - Arylzinc compounds, ArZnX, were conveniently prepared in high yields by the reaction of zinc powder with aryl iodides, which contain electron-withdrawing groups such as CO(2)CH(3), CN, Br, or CF(3) at the ortho-, meta- or para position, or electron-donating groups such as CH(3), OCH(3), or H, at 70 degrees C in THF, at 100 degrees C, or at 130 degrees C in diglyme, respectively. Pd(dba)(2) exhibited an outstanding efficient catalytic effect for the cross coupling of these ethereal solutions of ArZnX with allylic halides to afford a variety of functionalized allylbenzenes in high yields; the reactions were mostly carried out at 0 degrees C for 5-30 min in the presence of 5 mol % of catalyst. The conversion of the aryl iodides to allylbenzenes via two reactions could be accomplished in one pot. PMID- 12636379 TI - Selective synthesis and isolation of all possible conformational isomers of proximally para-disubstituted calix[4]arene. AB - All six possible conformational isomers of the proximally p-dibrominated calix[4]arene tetraalkyl ether, 1a-f*, were selectively synthesized by appropriate control of stereochemistry during di-O-alkylation reactions of 5,11 dibromocalix[4]arene syn-dialkyl ethers, namely, 5,11-dibromo-27,28-dihydroxy 25,26-dipropoxy-, 5,11-dibromo-25,26-dihydroxy-27,28-dipropoxy-, 5,11-dibromo 25,28-dihydroxy-26,27-propoxy-, and 5,11-dibromo-26,28-dihydroxy-25,27 dipropoxycalix[4]arenes. Their conformations were confirmed by (1)H and (13)C NMR spectroscopy and are cone for 1a (u(Br)(Pr), u(Br)(Pr), u(H)(Pr),u(H)(Pr)), partial cone for 1b (u(Br)(Pr), d(Br)(Pr), u(H)(Pr),u(H)(Pr)) and 1d (u(Br)(Pr), u(Br)(Pr), u(H)(Pr),d(H)(Pr)), 1,2-alternate for 1c (u(Br)(Pr), u(Br)(Pr), d(H)(Pr),d(H)(Pr)) and 1e (u(Br)(Pr), d(Br)(Pr), d(H)(Pr),u(H)(Pr)), and 1,3 alternate for 1f (u(Br)(Pr), d(Br)(Pr), u(H)(Pr),d(H)(Pr)). Although both 1c and 1e are in the 1,2-alternate conformation, the conformation of 1e was found to be strongly distorted and distinct from that of 1c. PMID- 12636381 TI - Formal total synthesis of salicylihalamides A and B. AB - An efficient synthesis of the macrolactone 3 of the salicylihalamides in 10 linear steps from alkene 6 is described. The key steps involved a Stille coupling between the chiral stannane 5 and benzyl bromide 4, which produced alkene 15 in good yield, and subsequent base-induced macrolactonization then gave compound 3. Macrolactone 3 was then converted into the known salicylihalamide A intermediate 18 in a three-step sequence. Compound 3 was also converted into another known salicylihalamide A and B intermediate 23 in a five-step sequence. PMID- 12636382 TI - Synthetic studies toward sarain A. Formation of the western macrocyclic ring. AB - Starting with the tricyclic core 2b, annulation to form the 13-membered western ring of sarain A has been achieved to afford the macrocycle 30a by initial construction of the sterically congested quaternary center at C-3, followed by elaboration of the C-3 side-chain and ring-closing olefin metathesis. Also included is a parallel conversion of tricycle 2c to macrocycle 30b containing a functionalized side-chain at N-1 suitable for attachment of the eastern macrocyclic ring. PMID- 12636383 TI - Synthesis of novel aromatic nitroxides as potential DNA intercalators. An EPR spectroscopical and DFT computational study. AB - The synthesis and electron paramagnetic resonance (EPR) spectroscopic properties of three novel aromatic nitroxides and potential DNA intercalators, the carbazole based 3,6-dimethylcarbazole-9-oxyl, as well as the acridane-based 9 acridanylidenemalonitrile-10-oxyl and 9-ethylacridanylidenecyanoacetate-10-oxyl, are described. The two acridane-based nitroxides can be isolated and are stable in solution as well as in the solid state for several days. Continuous wave X band EPR measurements and density functional theory (DFT) calculations demonstrated that the spin density is delocalized over the whole molecule in all three cases. Furthermore, the DFT calculations provided insight into the molecular and electronic structures of these nitroxides and yielded hyperfine coupling constants which are in very good agreement with the experimental data allowing therefore an unambiguous assignment of the hyperfine couplings. PMID- 12636384 TI - On the mechanism of the unexpected facile formation of meso-diacetate products in enzymatic acetylation of alkanediols. AB - The mechanism of the unexpected facile formation of meso-diacetate previously observed in the enzymatic resolution of dl/meso mixtures of 2,4-pentanediol and 2,5-hexanediol with Candida antarctica lipase B has been elucidated. It was found that the formation of meso-diacetate proceeds via different mechanisms for the two diols. Enzyme-catalyzed acylation of AcO-d(3) labeled (R)-monoacetates of meso-2,4-pentanediol and meso-2,5-hexanediol and analysis of the meso-diacetates obtained show that the former reaction proceeds via intramolecular acyl migration while the latter occurs via direct S-acylation of the alcohol. For the (R) monoacetate of (R,S)-2,4-pentanediol the intramolecular acyl migration was fast and therefore direct S-acylation by the external acyl donor is suppressed. For the hexanediol monoacetate the rate ratio (pseudo E value) between (5R,2R)- and (5R,2S)-5-acetoxy-2-hexanol was experimentally determined to be k(R,R)/k(R,S) = 25, which is about 10-20 times lower than the E value for 2-pentanol and 2 octanol. In a preliminary experiment it was demonstrated that facile acyl migration in the 1,3-diol derivative can be utilized to prepare syn-1,3 diacetoxynonane (>90% syn) in high enantioselectivity (>99% ee) via a chemoenzymatic dynamic kinetic asymmetric transformation of a meso/dl mixture of 1,3-nonanediol. PMID- 12636385 TI - Conjugate additions of o-iodoanilines and methyl anthranilates to acetylenic sulfones. A new route to quinolones including first syntheses of two alkaloids from the medicinal herb Ruta chalepensis. AB - A novel synthesis of 2-substituted 4-quinolones was developed on the basis of the conjugate additions of appropriately ortho-substituted anilines to acetylenic sulfones followed by intramolecular acylation of the corresponding sulfone stabilized anions. Conjugate additions of variously substituted anilines to alkyl substituted acetylenic sulfones generally proceeded slowly and in poor yield, especially when the aniline contained an electron-withdrawing substituent such as an ester group. In some cases, the reactions were enhanced by the presence of DMAP and the use of an excess of the sulfone in aqueous DMF. N-Formylanilines proved superior to free anilines. The products were either vinyl or allyl sulfones, depending on the conditions and the structure of the reactants. The acetylenic sulfone exists in equilibrium with its allenic and propargylic isomers under base-catalyzed conditions. Therefore, any of the three unsaturated sulfones can serve as the starting material for the conjugate additions. o-Iodoanilines proved superior to methyl anthranilate derivatives and underwent conjugate additions smoothly. The products were subjected to palladium-catalyzed carbonylation in methanol, and the resulting methyl esters were cyclized by treatment with strong bases such as LiHMDS or LiTMP, followed by reductive desulfonylation with aluminum amalgam. The resulting 2-substituted 4-quinolones included the naturally occurring medicinal compounds 1 and 2 and the O-methyl derivative 3. PMID- 12636386 TI - An extremely facile aza-Bergman rearrangement of sterically unencumbered acyclic 3-aza-3-ene-1,5-diynes. AB - The factors that affect the kinetics of the aza-Bergman cyclization of aza enediynes (C,N-dialkynyl imines) have not previously been elucidated. Here we report our kinetic studies of the aza-Bergman reactions of a series of 6 triisopropylsilyl and 6-unsubstituted 1-phenyl-4-aryl-3-aza-3-ene-1,4-diynes in which the aryl group is phenyl, o-(methoxy)phenyl, or p-(methoxy)phenyl. These aza-enediynes are prepared as single isomers in modest yield from the corresponding 1-aryl-3-(triisopropylsilyl)propynone oximes. These aza-enediynes undergo aza-Bergman reaction followed by a rapid retro-aza-Bergman cyclization to afford beta-alkynyl acrylonitrile products. In no case are products corresponding to trapping the intermediate 2,5-didehydropyridine diradical isolated. While the rate of aza-Bergman cyclization is not greatly affected by the nature of the 4 aryl substituent, the rate is very dependent on the nature of the 6-substituent. 1-Phenyl-4-aryl-3-aza-3-ene-1,5-diynes that lack a 6-substituent undergo aza Bergman cyclization spontaneously at 20 degrees C with first-order half-lives of 36-78 min. The effect of solvent on the kinetics of aza-Bergman cyclization of 1,4-diphenyl-3-aza-3-ene-1,5-diyne was investigated. The rate of this cyclization is solvent dependent, proceeding more rapidly in less polar solvents. PMID- 12636387 TI - Lithiation of 2-chloro- and 2-methoxypyridine with lithium dialkylamides: initial ortho-direction or subsequent lithium ortho-stabilization? AB - The lithiation pathway of 2-chloro and 2-methoxypyridine with LDA and LTMP has been investigated using deuterated probes. The availability of both H-6 and H-3 protons on the pyridine nucleus was found to be critical to ensure complete C-3 lithiation. We thus concluded that the C-3 lithiation was not a straightforward process. A mechanism involving precomplexation of lithium dialkylamides near the H-6 proton and formation of a 3,6-dilithio pyridine intermediate is proposed. PMID- 12636388 TI - Synthesis of polyhydroxycyclohexanes and relatives from (-)-quinic acid. AB - An efficient and versatile strategy for the synthesis of polyhydroxycyclohexanes and related compounds 3-6 is reported. The successful synthesis of these analogues has been achieved from a common intermediate, quinic acid derived lactone 2, rapidly accessible from cheap and commercially available (-)-quinic acid (1) as a chiral template. A practical route involving stereocontrolled epoxide formation and hydrolysis has been developed for the synthesis of 2,3 trans analogues 3 and 4. The preparation of the 2,3-cis analogues 5 and 6 has been realized by diasteroselective oxidation of a 5,6-double bond. PMID- 12636389 TI - Amino-zinc-enolate carbometalation reactions: application to ring closure of terminally substituted olefin for the asymmetric synthesis of cis- and trans-3 prolinoleucine. AB - The amino-zinc-enolate cyclization reaction is a straightforward route for the synthesis of 3-substituted prolines. As classical intramolecular carbometalation reactions, the applicability of the addition of zinc to a double bond was limited to a substrate in which the terminal alkene carbon was unsubstituted. Being interested in the synthesis of cis- and trans-3-prolinoleucine derivatives for our structure-activity relation (SAR) studies, we focused our effort on the preparation of these compounds by amino-zinc-enolate cyclization of terminally substituted double bonds. Herein we report that the attachment of an activating group such as cyclopropyl to the terminal olefin carbon allows the amino-zinc enolate cyclization of a terminally substituted alkene. The reaction is stereospecific, leading to a trans-3-substituted proline derivative, whereas a cis stereochemistry was observed with the amino-zinc-enolate cyclization of terminally nonsubstituted olefins. Absolute configurations obtained for the 3 prolinoleucine were established by X-ray analysis, NMR, and optical activity comparison of the cis and trans derivatives obtained by an unambiguous pathway. PMID- 12636390 TI - Effect of 9,10-cyclic acetal stereochemistry on feasible operation of the alpha ketol rearrangement in highly functionalized paclitaxel (Taxol) precursors. AB - The convergent, stereocontrolled synthesis of enantiopure stereoisomeric 9,10 cyclic acetals, whose designed role was to serve as potential precursors to Taxol, is reported. These advanced intermediates are multiply functionalized and carry a bridgehead alpha-ketol array which was key to isomerization into the proper framework. In agreement with relative strain energy values obtained by MM3 calculations, a dichotomy was observed between these two families. While the trans-fused acetals failed to undergo bridge migration, their cis counterparts did so efficiently. In fact, isomerization was sufficiently rapid that oxygenation at C2 was now precluded. The operation of several unusual transannular hydride shifts is also detailed. PMID- 12636391 TI - A delicate balance of energetics. Subtleties associated with alpha-ketol-based bridge migration to afford 9-keto-10beta-p-methoxybenzyloxytaxanes. AB - The feasibility of the title reaction has been pursued for the purpose of advancing a concise total synthesis of Taxol. Of the two closely related series examined, the first featured an exo-methylene group at C4. The second consisted of an alpha-epoxide at that site. Strikingly, the olefinic construct proved inert to attempted alpha-ketol rearrangement. In contrast, the oxiranyl derivative isomerized smoothly. The reaction sequence associated with arrival at taxane 18 is short (15 steps from a D-camphor derivative) and notably efficient. The thermodynamic issues that are raised by this investigation have been clarified by an assessment of molecular mechanics-derived (MM3) steric energy calculations. PMID- 12636392 TI - Chemical modification of a highly functionalized taxane. The consequences of an absent bridgehead double bond on oxetane D-ring construction. AB - An oxetane D-ring has been fused to the framework of the highly functionalized taxane 2. The synthetic route is based on a trimethylsilyl triflate-promoted epoxide-opening step, followed by stereocontrolled, regioreversed oxirane formation and reductive transposition of this intermediate with bis(cyclopentadienyl)titanium(III) chloride. This last key step provides for the convenient implementation of additional hydroxyl groups ultimately conducive to intramolecular S(N)2 reaction. Tangential features of the route outlined herein include specific rearrangement reactions and a retro-aldol cleavage of ring A. PMID- 12636393 TI - Protease-catalyzed peptide synthesis for the site-specific incorporation of alpha fluoroalkyl amino acids into peptides. AB - Substitution of native amino acids by fluoroalkyl analogues represents a new approach for the design of biologically active peptides with increased metabolic stability as well as defined secondary structure and provides a powerful label for spectroscopic investigations. Here, we introduce a methodology for the incorporation of sterically demanding C(alpha)-fluoroalkyl amino acids into the P(1) position of peptides catalyzed by the commercially available proteases trypsin and alpha-chymotrypsin. The combination of 4-guanidinophenyl ester of C(alpha)-fluoroalkyl amino acids as substrate mimetics with frozen-state reaction conditions provided the most efficient strategy for protease-catalyzed site specific introduction of this kind of nonnatural amino acids into peptide sequences. Consequently, a library of di-, tri-, and tetrapeptides containing alpha-methyl, alpha-difluoromethyl, and alpha-trifluoromethyl alanine, leucine, and phenylalanine in the P(1) position was synthesized catalyzed by trypsin as well as alpha-chymotrypsin. Trypsin was shown to be the more versatile protease. PMID- 12636394 TI - Palladium-catalyzed hydrocarbonation and hydroamination of 3,3 dihexylcyclopropene with pronucleophiles. AB - The reaction of 3,3-dihexylcyclopropene 1 with carbon and amine pronucleophiles 2 in the presence of palladium catalysts proceeded smoothly to give the corresponding hydrocarbonation products 3, allylated nucleophiles, in good to high yields. For example, in the presence of catalytic amounts of Pd(PPh(3))(4) and dppf, the reaction of 3,3-dihexylcyclopropene with ethyl 2-cyanopropionate and ethyl 2-cyanophenylacetate gave ethyl 2-cyano-2-methyl-4-undecenoate and ethyl 2-cyano-2-phenyl-4-undecenoate in 82 and 86% yield, respectively. PMID- 12636395 TI - Phase-transfer-catalyzed alkylation of guanidines by alkyl halides under biphasic conditions: a convenient protocol for the synthesis of highly functionalized guanidines. AB - An operationally straightforward and efficient method for the alkylation of carbamate-protected guanidines with various alkyl halides and mesylates is described. This protocol proceeds via deprotonation of the acidic N-carbamate hydrogen of the guanidine under biphasic conditions using a catalytic amount of a tetrabutylammonium salt as a phase-transfer catalyst. In this manner, highly functionalized guanidines can be obtained. The reaction is tolerant of a wide range of functional groups on both the alkyl halide and guanidine component. In addition, the reaction is sufficiently mild such that simple aqueous workup and filtration through a short silica gel column yields the substituted guanidines in high purity. In conjunction with the EDCI-mediated guanylation of disubstituted thioureas with amines, phase-transfer catalyzed alkylation of guanidines via a one-pot, three-component synthesis of substituted guanidines was achieved. PMID- 12636396 TI - Concerted rearrangement versus heterolytic cleavage in anionic [2,3]- and [3,3] sigmatropic shifts. A DFT study of relationships among anion stabilities, mechanisms, and rates. AB - The anionic [2,3] sigmatropic Wittig rearrangements of deprotonated 4-hetera-1 pentenes and the anionic [3,3] sigmatropic Cope rearrangements of 3-substituted 1,5-hexadienes were explored by using density functional theory calculations. While the deprotonated anionic 3-hydroxy-1,5 hexadiene (2a), 3-thiohydroxy-1,5 hexadiene (2c), and 3-formamidyl-1,5-hexadiene (2d) Cope substrates undergo concerted rearrangements, the deprotonated anionic 3-amino-1,5-hexadiene (2b) and 3-methyl-1,5-hexadiene (2e) Cope substrates follow nonconcerted cleavage/recombination pathways. We have also found that the gas-phase Wittig (1a), aza-Wittig (1b), and carba-Wittig (1c) reactions proceed via nonconcerted cleavage/recombination pathways. These results are compared with previous results on the Cope rearrangements of deprotonated anionic 3-hydroxy-1,5-hexadiene and 3 amino-1,5-hexadiene anions. A previously established model that heterolytic and homolytic bond dissociation energies can be used to predict how anionic amino- and oxy-Cope substrates will react is generalized to account for the reactivity of other Cope substrates as well as for the Wittig rearrangements. There is also a relationship between the basicity of the anionic substituent in the Cope rearrangement and the reaction pathway: the more basic the substituent anion, the less stable it is, and the more likely it is that cleavage will occur. A first step toward studying these reactions in solution was also taken by calculating energetics for some of the rearrangements with a lithium counterion present. PMID- 12636397 TI - Highly stereoselective hydroxy-directed Diels-Alder reaction. AB - The successful stereocontrol of the Diels-Alder reaction of semicyclic dienes possessing a secondary and tertiary allylic magnesium alkoxide alcohol functionality and activated dienophiles such as methyl acrylate, methacrolein, acrolein, and N-phenylmaleimide is described. PMID- 12636398 TI - Synthesis of all stereoisomers of sulfinylcalix[4]arenes. AB - All four stereoisomers of p-tert-butylsulfinylcalix[4]arene [4(rccc), 4(rcct), 4(rctt), and 4(rtct)], arising from the disposition of the four sulfinyl groups with respect to the mean plane of the four bridging sulfur atoms, have been prepared via oxidation of p-tert-butylthiacalix[4]arene (2) or its tetra-O-benzyl ethers 5 of defined conformations. Thus, treatment of 2 with 4.4 molar equiv of NaBO(3).4H(2)O gave the rtct and rctt isomers in 27% and 17% yields, respectively, while oxidation of cone 5 (5(C)) and partial cone 5 (5(PC)) proceeded stereoselectively to give, after debenzylation of the resulting tetrasulfoxides 12 and 15, the rccc and rcct isomers in 56% and 28% yields, respectively, based on 5. The sulfinylcalix[4]arenes 4 were treated with iodomethane in the presence of a base to give the corresponding tetramethyl ethers 16, the structures of which in regard to the disposition of the sulfinyl groups and the conformation of the phenol units were determined by X-ray crystallography. Also reported is the synthesis of all four conformational isomers of tetra-O-benzyl ether of 2 (5(C), 5(PC), 5(1,2-A), and 5(1,3-A)). PMID- 12636399 TI - Synthesis of 1,4-diazines by ring expansion of 1,3-diazines. AB - Fully saturated piperazin-3-one and quinoxalin-3-one derivatives were prepared by reactions of 2-anilino-2-ethoxy-3-oxothiobutanoic acid anilide with aliphatic 1,2 diamines. An unusual ring expansion of the intermediate 1,3-diazines leads to 1,4 diazines. Moreover, quinoxalin-3-one derivatives from chiral trans-1,2 diaminocyclohexane were obtained with diastereoselectivity >95%. PMID- 12636400 TI - A practical synthesis for the core structure of a family of selective prostaglandin D2 receptor antagonists. AB - A convergent synthesis was developed for the production of the core structure of prostaglandin D(2) receptor antagonists for the treatment of allergic rhinitis. The key steps in this synthesis were a highly diastereoselective alkylation of (+)-nopinone, a chemo- and stereoselective reduction of an oxime to an amine, and a well-controlled reduction of an aminoalkyne to a (Z)-olefin. PMID- 12636401 TI - Serine-cis-proline and serine-trans-proline isosteres: stereoselective synthesis of (Z)- and (E)-alkene mimics by Still-Wittig and Ireland-Claisen rearrangements. AB - Two new amide isosteres of Ser-cis-Pro and Ser-trans-Pro dipeptides were designed and stereoselectively synthesized to be incorporated into potential inhibitors of the phosphorylation-dependent peptidylprolyl isomerase Pin1, an essential regulator of the cell cycle. The cis mimic, the (Z)-alkene isomer, was formed through the use of a Still-Wittig [2,3]-sigmatropic rearrangement, while the trans mimic, the (E)-alkene, was synthesized through the use of an Ireland Claisen [3,3]-sigmatropic rearrangement. Starting from N-Boc-Ser(OBn)-N(OMe)Me, both mimics were synthesized in Boc-protected form suitable for peptide synthesis with an overall yield of 20% in 10 steps for the cis mimic and 13% in eight steps for the trans mimic. PMID- 12636402 TI - Activity-guided isolation of novel norwithanolides from depreasubtriflora with potential cancer chemopreventive activity. AB - Activity-monitored fractionation of a CHCl(3)-soluble extract of Deprea subtriflora using a quinone reductase induction assay led to the purification of subtrifloralactones A-J (1-10), 10 novel C-18 norwithanolides based on a new C(27) skeleton. These compounds were characterized by spectroscopic and chemical studies, and single-crystal X-ray diffraction analysis was used to confirm the structures of 1 and 4. Compounds 1-10 were evaluated for their cancer chemopreventive activity in terms of their ability to induce quinone reductase activity with cultured murine hepatoma cells, and compounds 1 and 6 were found to be highly effective. PMID- 12636403 TI - Fluorescent quenching of the 2-naphthoxide anion by aliphatic and aromatic halides. Mechanism and consequences of electron transfer reactions. AB - The fluorescent excited state of the 2-naphthoxide ion (1) is quenched by aliphatic and aromatic halides according to an electron-transfer mechanism, with generation of the corresponding alkyl and aryl radicals by a concerted or consecutive C-X bond fragmentation reaction. Whereas bromo- and iodobenzene follow a concerted ET mechanism (C-X, BDE control), 1-bromonaphthalene exhibits a stepwise process (pi LUMO control). The photoinduced reaction of anion 1 with 1 iodoadamantane (2) in DMSO affords substitution products on C3, C6, and C8, 1 adamantanol, 1-adamantyl 2-naphthyl ether, and adamantane (3.2, 13.2, 12.2, 2.8, 2.5, and 14.1% yields, respectively). A complex mixture is also observed in the photochemical reaction of neopentyl iodide (3) with anion 1, which renders substitution on C1, C3, C6, C8, and 2-naphthyl neopentyl ether (8.1, 1.3, 19.1, 31.1, and 2.8% yields, respectively). The absence of reaction in the dark and the inhibition of the photoinduced reaction by the presence of the radical traps di tert-butylnitroxide (DTBN) and 1,4-cyclohexadiene are evidence of a radical chain mechanism for these substitutions. On the other hand, only coupling at C1 is achieved by the photostimulated reaction of anion 1 with iodobenzene (5), to afford 41.9% of 1-phenyl-2-naphthol and 5.4% of disubstitution product. The regiochemistry of these reactions can be ascribed to steric hindrance and activation parameters. PMID- 12636404 TI - Molecular modeling of salt (lithium chloride) effects on the enantioselectivity of diethylzinc addition to benzaldehyde in the presence of chiral beta-amino alcohols. AB - Beta-amino alcohols (S,S,S)-1 and (R,R,S)-1, derived from cyclohexene oxide and containing alpha-phenylethyl auxiliaries, were examined as chiral promoters in the addition of diethylzinc to benzaldehyde. In agreement with literature precedent, the N-alpha-phenylethyl chiral auxiliary had no significant impact on enantioinduction, which is determined by the configuration of the framework's C(OH), with unlike induction. Contrary to some literature reports, stereoinduction by lithium salt derivatives of (S,S,S)-1 and (R,R,S)-1 was lower than that obtained with the free amino alcohol. Remarkable lithium chloride salt effects were observed in the reaction. In particular, an opposite chiral induction was found with (S,S,S)-1-Li(2) as ligand and in the presence of "inert" salt. N-Alkylated derivatives (S,S,S)-3-7 proved to be more efficient ligands, providing higher yields and enantioselectivities in the formation of carbinols (R)- or (S)-2. BP86/DN**//PM3 theoretical calculations proved remarkably successful in reproducing the experimental observations and permitted expansion of Noyori's catalytic cycle [J. Am. Chem. Soc. 1995, 117, 6327] to understand the relevant N-substitution and medium salt effects that determine the enantioselection in this catalytic asymmetric reaction. PMID- 12636405 TI - Enantioselective total syntheses of slagenins A-C and their antipodes. AB - Full details of the total syntheses of slagenins A-C (1a-c) and their antipodes (2a-c), novel bromopyrrole alkaloids with a unique tetrahydrofuro[2,3 d]imidazolidin-2-one moiety, are described in which their absolute stereochemistry was established. The key step in the syntheses involves the efficient condensation of dihydrofuran-3-one or glyoxal with urea to construct the slagenin bicycle core. PMID- 12636406 TI - Crowned dendrimers: pH-responsive pseudorotaxane formation. AB - With the end goal of incorporating the unique structural and physical properties of dendrimers into supramolecular assemblies, bis(m-phenylene)-32-crown-10 functionalized poly(propyleneimine) dendrimers of the first and third generations have been synthesized and their interaction with paraquat diol has been investigated. Using (1)H NMR, we determined that binding to the 4 or 16 crown ether sites occurred in an anti-cooperative fashion, most likely a result of steric influences. Upon protonation of the tertiary amines in the dendritic interior, binding became independent, i.e., statistical, and the average apparent association constant increased by nearly 5-fold; this effect is attributed to rigidification of the dendrimer, which makes its binding sites more accessible and less crowded. PMID- 12636407 TI - First experimental and theoretical evidence of a deactivating enone dienophile in the transannular Diels-Alder reaction. AB - A thorough study of the transannular Diels-Alder (TADA) reaction of trans-trans trans macrocyclic trienes was carried out. It led to a better understanding of various parameters that govern the TADA reaction in particular and the Diels Alder reaction in general. Thus, carbonyl activation of the dienophile is thoroughly discussed in light of new experimental and theoretical data. An enone dienophile is found to deactivate the reaction, although it remains planar at the transition state. This unusual result was discussed in terms of tether substituents that provoke destabilization of the transition state. PMID- 12636408 TI - New chiral pool approach to anthracyclinones. The stereoselective synthesis of idarubicinone. AB - In the present work, a new chiral pool approach has been developed for the synthesis of anthracyclinones. Thus, enone 8, readily available from l-rhamnose, has been converted via addition of 2,5-dimethoxybenzyllithium to the carbonyl group and a series of six reactions into a suitably protected aldehyde 21. The SnCl(4)-promoted stereospecific cyclization of the latter afforded enantiopure key intermediate 22. Silylation of benzylic hydroxyl of 21 followed by anodic oxidation and selective hydrolysis gave ketoacetals 25 and 26 to which 3-cyano 1(3H)-isobenzofuranone 27 was annelated. Removal of the isopropylidene group in the resulting 28, subsequent oxidation of the C(13) hydroxyl and full deprotection led to idarubicinone (4). PMID- 12636409 TI - Terpene biosynthesis in the nudibranch Doriopsilla areolata. AB - Biogenesis of the enantiomeric sesquiterpenes 1 and 5 of the marine nudibranch Doriopsilla areolata was investigated by feeding of [1-(13)C]glucose, [1,2 (13)C(2)]glucose, and [1,2-(13)C(2)]acetate. Evidence is presented that supports de novo origin of both compounds via mevalonic acid. PMID- 12636410 TI - Asymmetric synthesis of aziridine 2-phosphonates from enantiopure sulfinimines (N sulfinyl imines). Synthesis of alpha-amino phosphonates. AB - An aza-Darzens reaction, involving the addition of chloromethylphosphonate anions to enantiopure sulfinimines, has been developed for the asymmetric synthesis of aziridine 2-phosphonates. Best results involve cyclization of the syn and anti diastereomerically pure alpha-chloro-beta-amino phosphonates to cis- and trans-N sulfinyl aziridine 2-phosphonates, respectively, with n-BuLi. A transition-state hypothesis is proposed wherein the chloromethylphosphonate anion adds to the C-N bond on the side that is opposite the bulky p-tolyl sulfinyl group. The N sulfinyl group is easily removed by treatment with MeMgBr or TFA/MeOH, which affords the NH-aziridines in good yield. Using transfer hydrogenation conditions, the NH-aziridines were regioselectively opened to the corresponding enantiopure alpha-amino phosphonates without N-activation and in excellent yield. PMID- 12636411 TI - Intramolecular and intermolecular kinetic isotope effects (KIE) in the nitrosoarene ene reaction: experimental evidence for reversible intermediate formation. AB - The intramolecular and intermolecular kinetic isotope effects (KIE) have been determined for the nitrosoarene ene reaction with deuterium-stereolabeled 2,3 dimethyl-2-butenes (TME). trans-TME-d(6) (k(H)/k(D) = 3.0) and gem-TME-d(6) (k(H)/k(D) = 4.0) show large intramolecular primary isotope effects. In contrast, the intramolecular competition in cis-TME-d(6) (k(H)/k(D) = 1.5) and the intermolecular competition for the TME-d(0)/TME-d(12) pair (k(H)/k(D) = 1.98) show considerably smaller, but mechanistically significant kinetic isotope effects. The latter fact is rationalized in terms of reversible formation of a three-membered-ring intermediate, namely the aziridine N-oxide, or a similar unsymmetrical, polarized diradical in the first step of the reaction. Such reversibility has also been implied earlier for triazolinedione (TAD) and singlet oxygen ((1)O(2)) with deuterium-stereolabeled 2-butenes, but of the three enophiles, ArNO is the most sensitive toward reversibility, which is due to its moderate reactivity and its high steric demand. PMID- 12636413 TI - Synthesis of functionalized piperidinones. AB - A versatile, stereoselective synthesis of 5-hydroxypiperidinones with substituents at N1, C3, and C6 has been developed. The sequence involves ring closing metathesis of a diene amide and epoxidation of the resulting alkene, followed by base-mediated elimination, and finally hydrogenation. PMID- 12636412 TI - Simplifying oligosaccharide synthesis: efficient synthesis of lactosamine and siaylated lactosamine oligosaccharide donors. AB - A practical sequence is described for converting d-glucosamine into peracetylated Gal(beta-1,4)GlcNTroc(beta1-S)Ph and Neu5Ac(alpha-2,3)Gal(beta-1,4)GlcNTroc(beta1 S)Ph building blocks using a synthetic strategy based on chemoenzymatic oligosaccharide synthesis. The known trichloroethoxycarbonyl, N-Troc, protecting group was selected as a suitable protecting group for both enzymatic and chemical reaction conditions. These oligosaccharide building blocks proved effective donors for the beta-selective glycosylation of the unreactive OH-3 of a polymeric PEG-bound acceptor and for the axial OH-2 of a mannose acceptor in good yields. The resulting complex oligosaccharides are useful for vaccine and pharmaceutical applications. PMID- 12636414 TI - C2-Symmetric enantiopure ethanotethered bis(alpha,beta-butenolides) as templates for asymmetric synthesis. Application to the synthesis of (+)-grandisol. AB - Starting from D-mannitol, we have prepared several C(2)-symmetric ethanotethered bis(alpha,beta-butenolides) and studied their [2+2] photocycloaddition reaction with ethylene. The protective groups of the central diol unit have a noticeable influence on the facial selectivity of the cycloaddition, the bis(trimethylsilyloxy) derivatives showing the highest diastereoselectivity. A theoretical conformational analysis of the substrates in the ground state is in good agreement with the diastereofacial selectivity experimentally observed. The bis(photocycloadducts) have been converted into the enantiopure cyclobutanes formally derived from the photoreaction of ethylene with gamma-hydroxymethyl alpha,beta-butenolide, in which only a moderate facial selectivity had been previously found. As an application of these studies, we have developed a highly efficient and stereoselective synthesis of (+)-grandisol. PMID- 12636415 TI - Conformational analysis: crystallographic, NMR, and molecular mechanics studies of flexible sulfonic esters. AB - Two novel X-ray structures of the sulfonic ester derivatives 2-(6-iodo-1,3 benzodioxol-5-yl)ethyl 4-nitrobenzenesulfonate, 3, and 2-(6-iodo-1,3-benzodioxol 5-yl)ethyl 4-methylbenzenesulfonate, 4, have been obtained in a study aimed at analyzing the structures and conformations of sulfonic ester derivatives that are routinely used in alkaloid syntheses. The crystal structure of 4 is highly unusual, containing four independent molecules that belong to two distinct conformational types: (1) a hairpin conformation (stabilized mainly by intramolecular pi-stacking) and (2) a stepped conformation (stabilized mainly by intermolecular pi-stacking). Compound 3, on the other hand, crystallizes exclusively as the hairpin conformer. New MM+ force field parameters for sulfonic esters have been developed using the X-ray data, empirical rules, and DFT calculations to estimate the bond dipole parameters. Grid searches of conformational space for 3 and 4 using MM methods show that there are several gas phase conformations within 5 kcal/mol of the global minimum and that the lowest energy conformations (by approximately 4.6 kcal/mol) are of the hairpin type. Analysis of the MM conformational energies suggests that the dominant intramolecular interaction stabilizing the hairpin conformations of 3 and 4 is van der Waals attraction. Moreover, the lattice energies for packing the hairpin conformations of 3 and 4 are approximately 4 kcal/mol more favorable than for the stepped conformations. Various intermolecular interactions contribute to the complexity of the observed crystal structures of 3 and 4, including electrostatic attraction between O and I atoms in neighboring molecules. Langevin dynamics (LD) simulations at several temperatures (6.0 ns, friction coefficient = 2.5 ps(-1)) indicate that the conformational exchange rates are approximately 10(10)-10(11) s(-1) over the temperature range 213-400 K, accounting for the temperature independent (1)H NMR spectra of 3 and 4. PMID- 12636416 TI - Direct conversion of tert-beta-bromo alcohols to ketones with zinc sulfide and DMSO. AB - tert-Beta-bromo alcohols, derived from simple monoterpene hydrocarbons, react with zinc sulfide in dimethyl sulfoxide to afford saturated ketones as the major and hydroxy ketones as the minor products. The reaction involves initial nucleophilic attack by DMSO on the carbon attached to the halogen, which is assisted by electrophilic zinc sulfide. Subsequent Kornblum type oxidation yields the alpha-hydroxy ketone. On the other hand, abstraction of proton beta to the hydroxyl group followed by an attack of the neighboring hydroxyl moiety on the sulfur of the dimethylsulfoxonium intermediate and its subsequent collapse yields an enol, which tautomerizes to a saturated ketone. The latter pathway is predominantly followed. PMID- 12636417 TI - Directional synthesis of a dye-linked conducting homopolymer. AB - We report a new synthesis of a 10,20-diphenyl-5,15-bis(4-cyanophenyl)porphyrin (1) and its reduction to give 10,20-diphenyl-5,15-bis(4-formylphenyl)porphyrin (2). When the conducting polymer poly(2',5'-dioctyl-4,4' ' terphenylenecyanovinylene) was prepared in the presence of 2, we obtained a three domain structure, compound 3, consisting of two homopolymer blocks (J domains) with the porphyrin dye molecule (P domain) in the middle. The JPJ assembly was subjected to photophysical studies where we showed that the J domain could serve as a light-harvesting antenna for the P domain. PMID- 12636418 TI - Molecular recognition of fluoride anion: benzene-based tripodal imidazolium receptor. AB - A benzene-based tripodal imidazolium receptor utilizing the strong (C-H)(+)...X( ) hydrogen bonding interaction between imidazolium moieties and halide anions is extensively investigated both theoretically and experimentally. Ab initio calculations predict that this receptor has a very high affinity for fluoride ion (F(-)). The association constant and free energy gain of the N-butyl receptor 2 for F(-) in acetonitrile were measured to be 2.1 x 10(5) M(-1) and -7.25 kcal/mol, respectively, showing that the receptor has a high affinity for F(-) in highly polar organic solvents. PMID- 12636419 TI - Facile transformation of 2-azetidinones to 2-piperidones: application to the synthesis of the indolizidine skeleton and (8S,8aS)-perhydro-8-indolizinol. AB - The highly functionalized bicyclic lactam 7 was prepared from diolefinic-2 piperidone 18 by the use of ruthenium-catalyzed RCM, and in turn, 18 was derived via a two-carbon addition process from readily accessible 4-olefinic-2 azetidinone 13. Bicyclic lactams 7 and 20 could serve as potentially valuable intermediates for the chiral synthesis of various hydroxylated indolizidine alkaloids as exemplified by the synthesis of (8S,8aS)-perhydro-8-indolizinol 19. PMID- 12636420 TI - Synthesis of two fluoro analogues of the nicotinic acetylcholine receptor agonist UB-165. AB - Two racemic fluoropyridine analogues 4 and 5 of the potent nicotinic agonist UB 165 have been synthesized. Halogenated pyridines 7 and 12 provided the organometallic reagents needed for the Negishi and Suzuki coupling reactions used for the preparation of 4 and 5, and the N-vinyloxycarbonyl protecting group of 8 and 15 was cleaved using a novel trifluoroacetic acid-mediated deprotection protocol. Analogue 4 retained high binding affinity at rat brain alpha4beta2 and alpha7 nicotinic receptors. PMID- 12636421 TI - Practical synthesis of optically active alpha,alpha-disubstituted malonamic acids through asymmetric hydrolysis of malonamide derivatives with Rhodococcus sp. CGMCC 0497. AB - A variety of alpha,alpha-disubstituted malonamides undergo enantioselective hydrolysis with Rhodococcus sp. CGMCC 0497 to give challenging enantiopure alpha,alpha-disubstituted malonamic acids with up to >99% enantiomeric excesses and 98% chemical yields. The enantioselectivity originated from the effects of a highly enantioselective amidase. The products could be converted to valuable (R)- or (S)-alpha,alpha-dialkylated amino acids after routine conversions. PMID- 12636422 TI - Photochemical protection of amines with Cbz and Fmoc groups. AB - The photochemical conversion of amines into carbamates was achieved using N-Cbz-, N-Fmoc-, and N-Boc-5,7-dinitroindolines. This reaction allows the protection of amines in neutral medium. Primary and unhindered secondary amines were protected to yield their benzyloxycarbonyl- and 9-fluorenylmethoxycarbonyl derivatives efficiently, whereas bulky amines or anilines gave low yields or no product. On the other hand, the formation of N-Boc compounds, although possible, proceeded only with low yields. PMID- 12636423 TI - Optimization of the synthesis of symmetric aromatic tri- and tetrasulfides. AB - The reaction of aromatic thiols with sulfur dichloride and sulfur monochloride to form the corresponding aromatic trisulfides, 2a-d, and tetrasulfides, 3a-d, has been optimized with respect to yield and purity. The use of pyridine as an amine base and the use of freshly distilled sulfur monochloride (S(2)Cl(2)) serve as important alterations to the synthetic method. Their physical properties have been characterized, revealing some discrepancies with the literature. PMID- 12636424 TI - Ru-catalyzed highly enantioselective hydrogenation of beta-alkyl-substituted beta (acylamino)acrylates. AB - Highly enantioselective hydrogenation of beta-alkyl-substituted (E)-beta (acylamino)-acrylates catalyzed by Ru((R)-Xyl-P-Phos)(C(6)H(6))Cl(2) complex (cat. 1c) was achieved in up to 99.7% ee. Moderate to good enantioselectivities in the hydrogenation of corresponding (Z)-isomers in the presence of [Rh((R)-Xyl P-Phos)(COD)]BF(4) (cat. 2c) were also obtained. The results demonstrated that the electronic and steric properties of the dipyridylphosphine ligands as well as the different transition metal ions have significant influences on the catalytic properties in the hydrogenation of beta-(acylamino)acrylates. PMID- 12636425 TI - Enantioselective fluorination mediated by N-fluoroammonium salts of cinchona alkaloids: first enantioselective synthesis of BMS-204352 (MaxiPost). AB - We have employed a cinchona alkaloid/Selectfluor-mediated enantioselective fluorination of the oxindole 2 to achieve the first enantioslective synthesis of BMS-204352 (MaxiPost, S-1), an effective opener of maxi-K channels. Fluorination occurred to produce S-1 with 84% ee using the bis-cinchona alkaloid (DHQ)(2)AQN. Recrystallization produced enantiomerically pure (>99% ee) product. Quinidine mediated fluorination of 2 gave the (R)-antipode of 1 with 68% ee. PMID- 12636426 TI - Direct amination of 1-substituted 3,5-dinitrobenzenes by 1,1,1 trimethylhydrazinium iodide. AB - The amination of 1-X-3,5-dinitrobenzenes via the vicarious nucleophilic substitution of hydrogen (VNS) with 1,1,1-trimethylhydrazinium iodide (TMHI) in the presence of t-BuOK or NaOMe in DMSO was studied. It was observed (when X = OMe, OCH(2)CF(3), OCH(2)CF(2)CF(2)H, OPh) that the amination occurs regioselectively (ratio of ortho/para-isomers is approximately 9:1) and with high yield. For X = SPh or SCH(2)Ph, the reaction proceeded with a low yield (less than 20%), with a ratio of ortho/para-isomers approximately 1:1. For X = PhSO(2) and 2 equiv of TMHI, a double amination occurs and 2,4-diamino-3,5-dinitro-1 phenylsulfonylbenzene predominates in the mixture of isomers. Under the same conditions, 1,3,5-trinitrobenzene undergoes a double amination to yield 2,4 diamino-1,3,5-trinitrobenzene. A proposed mechanism for this reaction is discussed. PMID- 12636427 TI - Studies on quinazolines. 11. Intramolecular imidate-amide rearrangement of 2 substituted 4-(omega-chloroalkoxy)quinazoline derivatives. 1,3 -O --> N shift of chloroalkyl groups via cyclic 1,3-azaoxonium intermediates. AB - The omega-chloroalkylation of 2-substituted quinazolin-4(3H)-one derivatives 1 and 2 with Br-(CH(2))(n)-Cl (n = 2-4) and the intramolecular imidate-amide rearrangement of the alkylated products are described. At room temperature, the 2 phenyl substituent promoted O-alkylation, whereas the less steric 2-benzyl group led to a higher ratio of N-alkylation. The investigation of the O-alkylated products, 4-omega-chloroalkoxyquinazolines, revealed that the migration of omega chloroethyl and omega-chloropropyl groups from oxygen to nitrogen should be intramolecular via five- and six-membered cyclic 1,3-azaoxonium intermediates, respectively. Competition between rearrangement and nucleophilic substitution results in the formation of 7a,b and 8a,b from the nucleophilic substitution of 4a,b and 6a,b, respectively. PMID- 12636429 TI - Highly efficient 1,4-addition of 1,3-diesters to conjugated enones by In/TMSCl. AB - Organoindium reagents derived from indium and diethyl bromomalonates were added to a wide range of conjugated enones in a 1,4-fashion in the presence of TMSCl under mild conditions, and corresponding oxo-1,3-diesters were obtained in good to excellent yields. PMID- 12636428 TI - Synthesis of a C-glycoside analogue of beta-D-galactosylthreonine. AB - A C-linked analogue of beta-D-galactosylthreonine has been prepared from 2,3,4,6 tetra-O-benzyl-D-galactopyranolactone (1) in 14 steps. Three stereogenic centers were created during the synthesis, with the anomeric center of the C-glycoside being generated first by addition of a Grignard reagent to 1 and subsequent reduction of the intermediate hemiacetal with triethylsilane. The two stereogenic centers in the threonine moiety were both established by alkylation of Evans' chiral N-acyloxazolidinone enolates. PMID- 12636430 TI - Divergent synthesis of D-erythro-sphingosine, L-threo-sphingosine, and their regioisomers. AB - Starting from a vinyl epoxide, a divergent synthesis of four sphingosine isomers is described. The remaining four isomers can easily be synthesized using the same methodology. Although numerous syntheses of sphingosine have been published, this is the first general route leading to all eight isomers in this important compound class. The synthetic strategy relies on regioselective opening of a vinyl epoxide and a vinylaziridine in the allylic position. PMID- 12636431 TI - Copper-catalyzed regioselective allylic substitution reactions with indium organometallics. AB - The first nucleophilic allylic substitution reactions of triorganoindium compounds with allylic halides and phosphates are reported. The reactions of trialkyl- and triarylindium reagents with cinnamyl and geranyl halides and phosphates, with the aid of copper catalysis [Cu(OTf)(2)/P(OEt)(3)], are described. In general, the reaction proceeds efficiently to give good yields and regioselectively to afford the S(N)2' product. PMID- 12636432 TI - A one-pot strategy for synthesis of 5-O-(alpha-D-arabinofuranosyl)-6-O-(beta-D galactofuranosyl)-D-galactofuranose present in motif E of the Mycobacterium tuberculosis cell wall. AB - 5-O-(Alpha-D-arabinofuranosyl)-6-O-(beta-D-galactofuranosyl)-D-galactofuranose 6 present in motif E of the Macobacterium tuberculosis cell wall has been regio- and stereospecifically synthesized using 3-O-benzoyl-1,2-O-isopropylidine-alpha-D galactofuranose (10) as the glycosyl acceptor by the trichloroacetamidate method in a one-pot manner. The diol glycosyl acceptor 10 was smoothly derived from 1,2:5,6-di-O-isopropylidene-alpha-D-galactofuranose (8) by 3-O-benzoylation and then selective 5,6-O-deacetonation. The preparation of 8 was greatly improved by increasing the ratio of DMF to acetone and using a solid-supported catalyst. PMID- 12636433 TI - [Bmim]PF(6): a novel and recyclable ionic liquid for conversion of oxiranes to thiiranes in aqueous media. AB - A variety of epoxides respond rapidly with potassium thiocyanate in [bmim]PF(6) H(2)O (2:1) solvent system at room temperature under mild and convenient conditions to produce the corresponding thiiranes in high to quantitative yields. Enhanced rates, improved yields, and recyclability of ionic liquids are the remarkable features observed in ionic liquids (ILs). The use of ionic liquids for this transformation avoids the use of heavy metal halides as promoters and chlorinated hydrocarbons as solvents. The ionic liquid was recycled in five to six subsequent runs with gradual decrease in activity. PMID- 12636437 TI - A piece of my mind. Dumbing down. PMID- 12636438 TI - Tensions remain over tPA for stroke. PMID- 12636439 TI - Hemochromatosis patients are untapped source of blood as war, shortages loom. PMID- 12636440 TI - Trials planned for testing nonsurgical approach to replacing heart valves. PMID- 12636441 TI - From the Centers for Disease Control and Prevention. Fatal degenerative neurologic illnesses in men who participated in wild game feasts--Wisconsin, 2002. PMID- 12636445 TI - From the Centers for Disease Control and Prevention. Outbreak of group A streptococcal pneumonia among Marine Corps Recruits--California, November 1 December 20, 2002. PMID- 12636446 TI - Diuretics in critically ill patients with acute renal failure. PMID- 12636447 TI - Diuretics in critically ill patients with acute renal failure. PMID- 12636448 TI - Diuretics in critically ill patients with acute renal failure. PMID- 12636449 TI - Diuretics in critically ill patients with acute renal failure. PMID- 12636450 TI - Diuretics in critically ill patients with acute renal failure. PMID- 12636451 TI - Sestamibi imaging to triage patients with acute chest pain. PMID- 12636452 TI - Sestamibi imaging to triage patients with acute chest pain. PMID- 12636453 TI - From the Centers for Disease Control and Prevention. Public health and aging: trends in aging--United States and worldwide. PMID- 12636454 TI - From the Centers for Disease Control and Prevention. Public health dispatch: outbreaks of community-associated methicillin-resistant Staphylococcus aureus skin infections--Los Angeles County, California, 2002-2003. PMID- 12636455 TI - Recurrent otitis media in children. PMID- 12636456 TI - Recurrent otitis media in children. PMID- 12636457 TI - Recurrent otitis media in children. PMID- 12636458 TI - Recurrent otitis media in children. PMID- 12636461 TI - Delaying defibrillation to give basic cardiopulmonary resuscitation to patients with out-of-hospital ventricular fibrillation: a randomized trial. AB - CONTEXT: Defibrillation as soon as possible is standard treatment for patients with ventricular fibrillation. A nonrandomized study indicates that after a few minutes of ventricular fibrillation, delaying defibrillation to give cardiopulmonary resuscitation (CPR) first might improve the outcome. OBJECTIVE: To determine the effects of CPR before defibrillation on outcome in patients with ventricular fibrillation and with response times either up to or longer than 5 minutes. DESIGN, SETTING, AND PATIENTS: Randomized trial of 200 patients with out of-hospital ventricular fibrillation in Oslo, Norway, between June 1998 and May 2001. Patients received either standard care with immediate defibrillation (n = 96) or CPR first with 3 minutes of basic CPR by ambulance personnel prior to defibrillation (n = 104). If initial defibrillation was unsuccessful, the standard group received 1 minute of CPR before additional defibrillation attempts compared with 3 minutes in the CPR first group. MAIN OUTCOME MEASURE: Primary end point was survival to hospital discharge. Secondary end points were hospital admission with return of spontaneous circulation (ROSC), 1-year survival, and neurological outcome. A prespecified analysis examined subgroups with response times either up to or longer than 5 minutes. RESULTS: In the standard group, 14 (15%) of 96 patients survived to hospital discharge vs 23 (22%) of 104 in the CPR first group (P =.17). There were no differences in ROSC rates between the standard group (56% [58/104]) and the CPR first group (46% [44/96]; P =.16); or in 1-year survival (20% [21/104] and 15% [14/96], respectively; P =.30). In subgroup analysis for patients with ambulance response times of either up to 5 minutes or shorter, there were no differences in any outcome variables between the CPR first group (n = 40) and the standard group (n = 41). For patients with response intervals of longer than 5 minutes, more patients achieved ROSC in the CPR first group (58% [37/64]) compared with the standard group (38% [21/55]; odds ratio [OR], 2.22; 95% confidence interval [CI], 1.06-4.63; P =.04); survival to hospital discharge (22% [14/64] vs 4% [2/55]; OR, 7.42; 95% CI, 1.61-34.3; P =.006); and 1-year survival (20% [13/64] vs 4% [2/55]; OR, 6.76; 95% CI, 1.42 31.4; P =.01). Thirty-three (89%) of 37 patients who survived to hospital discharge had no or minor reductions in neurological status with no difference between the groups. CONCLUSIONS: Compared with standard care for ventricular fibrillation, CPR first prior to defibrillation offered no advantage in improving outcomes for this entire study population or for patients with ambulance response times shorter than 5 minutes. However, the patients with ventricular fibrillation and ambulance response intervals longer than 5 minutes had better outcomes with CPR first before defibrillation was attempted. These results require confirmation in additional randomized trials. PMID- 12636462 TI - Cognitive behavioral therapy and aerobic exercise for Gulf War veterans' illnesses: a randomized controlled trial. AB - CONTEXT: Gulf War veterans' illnesses (GWVI), multisymptom illnesses characterized by persistent pain, fatigue, and cognitive symptoms, have been reported by many Gulf War veterans. There are currently no effective therapies available to treat GWVI. OBJECTIVE: To compare the effectiveness of cognitive behavioral therapy (CBT), exercise, and the combination of both for improving physical functioning and reducing the symptoms of GWVI. DESIGN, SETTING, AND PATIENTS: Randomized controlled 2 x 2 factorial trial conducted from April 1999 to September 2001 among 1092 Gulf War veterans who reported at least 2 of 3 symptom types (fatigue, pain, and cognitive) for more than 6 months and at the time of screening. Treatment assignment was unmasked except for a masked assessor of study outcomes at each clinical site (18 Department of Veterans Affairs [VA] and 2 Department of Defense [DOD] medical centers). INTERVENTIONS: Veterans were randomly assigned to receive usual care (n = 271), consisting of any and all care received from inside or outside the VA or DOD health care systems; CBT plus usual care (n = 286); exercise plus usual care (n = 269); or CBT plus exercise plus usual care (n = 266). Exercise sessions were 60 minutes and CBT sessions were 60 to 90 minutes; both met weekly for 12 weeks. MAIN OUTCOME MEASURES: The primary end point was a 7-point or greater increase (improvement) on the Physical Component Summary scale of the Veterans Short Form 36-Item Health Survey at 12 months. Secondary outcomes were standardized measures of pain, fatigue, cognitive symptoms, distress, and mental health functioning. Participants were evaluated at baseline and at 3, 6, and 12 months. RESULTS: The percentage of veterans with improvement in physical function at 1 year was 11.5% for usual care, 11.7% for exercise alone, 18.4% for CBT plus exercise, and 18.5% for CBT alone. The adjusted odds ratios (OR) for improvement in exercise, CBT, and exercise plus CBT vs usual care were 1.07 (95% confidence interval [CI], 0.63-1.82), 1.72 (95% CI, 0.91-3.23), and 1.84 (95% CI, 0.95-3.55), respectively. The OR for the overall (marginal) effect of receiving CBT (n = 552) vs no CBT (n = 535) was 1.71 (95% CI, 1.15-2.53) and for exercise (n = 531) vs no exercise (n = 556) was 1.07 (95% CI, 0.76-1.50). For secondary outcomes, exercise alone or in combination with CBT significantly improved fatigue, distress, cognitive symptoms, and mental health functioning, while CBT alone significantly improved cognitive symptoms and mental health functioning. Neither treatment had a significant impact on pain. CONCLUSION: Our results suggest that CBT and/or exercise can provide modest relief for some of the symptoms of chronic multisymptom illnesses such as GWVI. PMID- 12636463 TI - Prospective study of alcohol consumption and risk of dementia in older adults. AB - CONTEXT: Alcohol consumption has been associated with complex changes in cerebral vasculature and structure in older adults. How alcohol consumption affects the incidence of dementia is less clear. OBJECTIVE: To determine the prospective relationship of alcohol consumption and risk of dementia among older adults. DESIGN, SETTING, AND PARTICIPANTS: Nested case-control study of 373 cases with incident dementia and 373 controls who were among 5888 adults aged 65 years and older who participated in the Cardiovascular Health Study, a prospective, population-based cohort study in 4 US communities. The controls were frequency matched on age, death before 1999, and their attendance of a 1998-1999 clinic. Participants in this study underwent magnetic resonance imaging (MRI) of the brain and cognitive testing between 1992 and 1994 and were followed up until 1999. MAIN OUTCOME MEASURES: Odds of incident dementia, ascertained by detailed neurological and neuropsychological examinations according to average alcohol consumption, assessed by self-reported intake of beer, wine, and liquor at 2 visits prior to the date of the MRI. RESULTS: Compared with abstention, the adjusted odds for dementia among those whose weekly alcohol consumption was less than 1 drink were 0.65 (95% confidence interval [CI], 0.41-1.02); 1 to 6 drinks, 0.46 (95% CI, 0.27-0.77); 7 to 13 drinks, 0.69 (95% CI, 0.37-1.31); and 14 or more drinks, 1.22 (95% CI, 0.60-2.49; P for quadratic term =.001). A trend toward greater odds of dementia associated with heavier alcohol consumption was most apparent among men and participants with an apolipoprotein E epsilon4 allele. We found generally similar relationships of alcohol use with Alzheimer disease and vascular dementia. CONCLUSIONS: Compared with abstention, consumption of 1 to 6 drinks weekly is associated with a lower risk of incident dementia among older adults. PMID- 12636464 TI - Screening men for prostate and colorectal cancer in the United States: does practice reflect the evidence? AB - CONTEXT: The debate about the efficacy of prostate-specific antigen (PSA) screening for prostate cancer has received substantial attention in the medical literature and the media, but the extent to which men are actually screened is unknown. If practice were evidence-based, PSA screening would be less common among men than colorectal cancer screening, a preventive service of broad acceptance and proven efficacy. OBJECTIVE: To compare the prevalences of PSA and colorectal cancer screening among US men. DESIGN, SETTING, AND POPULATION: The 2001 Behavioral Risk Factor Surveillance System, an annual population-based telephone survey of US adults conducted by the Centers for Disease Control and Prevention, was used to gather data on a representative sample of men aged 40 years or older from all 50 states and the District of Columbia (n = 49 315). MAIN OUTCOME MEASURES: Proportions of men ever screened and up to date on screening for prostate cancer (with PSA testing) and colorectal cancer (with fecal occult blood testing, flexible sigmoidoscopy, or colonoscopy). RESULTS: Overall, men are more likely to report having ever been screened for prostate cancer than for colorectal cancer; 75% of those aged 50 years or older have had a PSA test vs 63% for any colorectal cancer test (risk ratio [RR], 1.20; 95% confidence interval [CI], 1.18-1.21). Up-to-date PSA screening is also more common than colorectal cancer screening for men of all ages. Among men aged 50 to 69 years (those for whom there is the greatest consensus in favor of screening), 54% reported an up to-date PSA screen, while 45% reported up-to-date testing for colorectal cancer (RR, 1.19; 95% CI, 1.16-1.21). In state-level analyses of this age group, men were significantly more likely to be up to date on prostate cancer screening compared with colorectal cancer screening in 27 states, while up-to-date colorectal cancer screening was more common in only 1 state. CONCLUSION: Among men in the United States, prostate cancer screening is more common than colorectal cancer screening. Physicians should ensure that men who choose to be screened for cancer are aware of the known mortality benefit of colorectal cancer screening and the uncertain benefits of screening for prostate cancer. PMID- 12636465 TI - Trends in incidence rates of invasive lobular and ductal breast carcinoma. AB - CONTEXT: Research has suggested that use of combined estrogen and progestin hormone replacement therapy (CHRT) increases breast cancer risk and that CHRT use is more strongly associated with the risk of invasive lobular breast carcinoma than that of invasive ductal carcinoma. Lobular carcinoma is less common than ductal carcinoma but can be more difficult to diagnose because of its subtle elusive infiltrative pattern. OBJECTIVE: To evaluate trends in invasive lobular and ductal carcinoma incidence rates from 1987 through 1999, during which time use of CHRT increased in the United States. DESIGN: Descriptive epidemiologic study. SETTING: Nine cancer registries that participate in the Surveillance, Epidemiology, and End Results (SEER) program of the National Cancer Institute and that cover Atlanta, Ga; Detroit, Mich; San Francisco-Oakland, Calif; Seattle, Wash; and Connecticut, Hawaii, Iowa, New Mexico, and Utah. POPULATION: Women 30 years of age and older residing in the areas covered by the 9 SEER registries. MAIN OUTCOME MEASURES: Proportional changes in incidence rates of invasive lobular and ductal carcinoma among women with no prior history of breast cancer. RESULTS: A total of 190 458 women were included in this analysis who were identified through the registries as having invasive breast cancer; 7682 of the 198 140 potentially eligible women (ie, those identified as not having in situ breast cancer) were excluded from this analysis because stage of cancer was unknown. Invasive breast cancer incidence rates adjusted for age and for SEER historic stage increased 1.04-fold (95% confidence interval [CI], 1.004-1.07) from 1987-1999 (206.7/100 000 to 214.1/100 000, age-adjusted). However, incidence rates of tumors classified as lobular increased 1.52-fold (95% CI, 1.42-1.63), and those classified as mixed ductal-lobular increased 1.96-fold (95% CI, 1.80 2.14); rates of these types combined increased 1.65-fold (95% CI, 1.55-1.78) (19.8/100 000 to 33.4/100 000, age-adjusted). In contrast, ductal carcinoma rates remained largely constant (153.8/100 000 to 155.3/100 000, age-adjusted; proportional change, 1.03 [95% CI, 0.99-1.06]). The proportion of breast cancers with a lobular component increased from 9.5% in 1987 to 15.6% in 1999. CONCLUSIONS: Ductal carcinoma incidence rates remained essentially constant from 1987-1999 while lobular carcinoma rates increased steadily. This increase presents a clinical challenge given that lobular carcinoma is more difficult to detect than ductal carcinoma by both physical examination and mammography. PMID- 12636466 TI - Newborns killed or left to die by a parent: a population-based study. AB - CONTEXT: Interest in the discarding or killing of newborns by parents has increased due to wide news coverage and efforts by states to provide Safe Haven legislation to combat the problem. OBJECTIVE: To describe the characteristics of these cases in North Carolina. DESIGN, SETTING, AND POPULATION: Case series derived from data on all deaths among liveborn infants 0 to 4 days of age reported to the North Carolina medical examiner from 1985 through 2000. MAIN OUTCOME MEASURES: Incidence of newborns known to have been killed or discarded by a parent; epidemiological characteristics of newborns and parents. RESULTS: There were 34 newborns known to have been killed or discarded by a parent, comprising 0.002% of all liveborn infants during the 16-year study period, giving a rate of 2.1 per 100 000 per year. A total of 58.8% were male, 41.1% were white, and 52.9% were black. For 29 cases, the perpetrator was determined to be the mother. Among mothers, 50% were single and 20.6% were married (marital status of the remainder was unknown). Thirty-five percent had had other children. Eight mothers (23.5%) were known to have received some prenatal care. The mean age of the mothers was 19.1 years (range, 14-35 years) and more than half were aged 18 years or older. The most common causes of death were asphyxiation/strangulation (41.1%) and drowning (26.5%). CONCLUSIONS: In North Carolina, at least 2.1 per 100 000 newborns are known to be killed or left to die per year, usually by their mothers. It is unknown how many of these deaths might be prevented by Safe Haven laws. Efforts to educate the public about these laws need to target the general public. Where resources are limited, the focus should be on on adolescent pregnancy prevention programs, young adults, prenatal care clinics, and married women. PMID- 12636467 TI - Headache assessment and management. PMID- 12636468 TI - Priming the pump--can delaying defibrillation improve survival after sudden cardiac death? PMID- 12636469 TI - Treating Gulf War veterans' illnesses--are more focused studies needed? PMID- 12636471 TI - JAMA patient page. Headaches. PMID- 12636472 TI - Towards deterministic equations for Levy walks: the fractional material derivative. AB - Levy walks are random processes with an underlying spatiotemporal coupling. This coupling penalizes long jumps, and therefore Levy walks give a proper stochastic description for a particle's motion with broad jump length distribution. We derive a generalized dynamical formulation for Levy walks, in which the fractional equivalent of the material derivative occurs. Our approach is expected to be useful for the dynamical formulation of Levy walks in an external force field or in phase space, for which the description in terms of the continuous time random walk or its corresponding generalized master equation are less well suited. PMID- 12636473 TI - Bifurcation, bimodality, and finite variance in confined Levy flights. AB - We investigate the statistical behavior of Levy flights confined in a symmetric, quartic potential well U(x) proportional, variant x(4). At stationarity, the probability density function features a distinct bimodal shape and decays with power-law tails which are steep enough to give rise to a finite variance, in contrast to free Levy flights. From a delta-initial condition, a bifurcation of the unimodal state is observed at t(c)>0. The nonlinear oscillator with potential U(x)=ax(2)/2+bx(4)/4, a,b>0, shows a crossover from unimodal to bimodal behavior at stationarity, depending on the ratio a/b. PMID- 12636474 TI - Universality classification of restricted solid-on-solid type surface growth models. AB - We consider the restricted solid-on-solid (RSOS) type surface growth models and classify them into dynamic universality classes according to their symmetry and conservation law. Four groups of RSOS-type microscopic models--asymmetric (A), asymmetric-conserved (AC), symmetric (S), and symmetric-conserved (SC) groups- are introduced and the corresponding stochastic differential equations (SDEs) are derived. Analyzing these SDEs using dynamic renormalization group theory, we confirm the previous results that A-RSOS, AC-RSOS, and S-RSOS groups belong to the Kardar-Parisi-Zhang class, the Villain-Lai-Das Sarma class, and the Edwards Wilkinson class, respectively. We also find that SC-RSOS group belongs to a new universality class featuring the conserved-cubic nonlinearity. PMID- 12636475 TI - Repulsive step potential: a model for a liquid-liquid phase transition. AB - In the framework of the perturbation theory for fluids we study the model with the isotropic repulsive step potential which in addition to the hard core has a repulsive soft core of a larger radius. It is shown that this purely repulsive potential is sufficient to explain a liquid-liquid phase transition and liquid anomalies. PMID- 12636476 TI - Equation of state of supercooled water from the sedimentation profile. AB - To study the coexistence of two liquid states of water within one simulation box, we implement an equilibrium sedimentation method--which involves applying a gravitational field to the system and measuring or calculating the resulting density profile in equilibrium. We simulate a system of particles interacting via the Stillinger-2 (ST2) potential, a model for water. We detect the coexistence of two liquid phases at low temperature. PMID- 12636477 TI - Velocity distribution of inelastic granular gas in a homogeneous cooling state. AB - The velocity distribution of inelastic granular gas is examined numerically on a two-dimensional hard disk system in nearly elastic regime using molecular dynamical simulations. The system is prepared initially in the equilibrium state with the Maxwell-Boltzmann distribution, then after several inelastic collisions per particle, the system falls in the state that the Boltzmann's equation predicts with the stationary form of velocity distribution. It turns out, however, that due to the velocity correlation the form of the distribution function does not stay time independent, but gradually returns to the Maxwellian immediately after the initial transient till the clustering instability sets in. It shows that, even in the homogeneous cooling state (Haff state), where the energy decays exponentially as a function of collision number, the velocity correlation in the inelastic system invalidates the assumption of molecular chaos and the prediction of the Boltzmann's equation fails. PMID- 12636478 TI - Coarsening of granular clusters: Two types of scaling behaviors. AB - We report on an experimental study of small cluster dynamics during the coarsening process in driven granular submonolayers of 120-microm bronze particles. The techniques of electrostatic and vertical mechanical vibration were employed to excite the granular gas. We measure the scaling exponent for the evaporation of small clusters during coarsening. It was found that the surface area of small clusters S vs time t behaves as S to (t(0)-t)(2/3) for lower frequencies and S to (t(0)-t) for higher frequencies. We argue that the change in the scaling exponent is related to the transition from three-dimensional (3D) to 2D character of motion in the granular gas. PMID- 12636479 TI - Self-similar dynamic quasi-two-dimensional sand fronts. AB - We report on a study of advancing quasi-two-dimensional sand fronts on an inclined flat and thin strip confined between two vertical plates. These fronts form when a thin initial stream of sand running down the flat obstacle gets trapped at some distance from the injection point. Right after this trapping, the front starts to advance upstream and grow in time. The shapes at successive times are found to be self-similar in time. The stability conditions for the obtained fronts are also outlined. A simple model for interface dynamics gives reasonable predictions for the observed shapes. PMID- 12636480 TI - Mounding instability and incoherent surface kinetics. AB - Mounding instability in a conserved growth from vapor is analyzed within the framework of adatom kinetics on the growing surface. The analysis shows that depending on the local structure of the surface, kinetics of adatoms may vary, leading to disjoint regions in the sense of a continuum description. This is manifested particularly under the conditions of instability. Mounds grow on these disjoint regions and their lateral growth is governed by the flux of adatoms hopping across the steps in the downward direction. Asymptotically ln t dependence is expected in (1+1) dimensions. Simulation results confirm the prediction. Growth in (2+1) dimensions is also discussed. PMID- 12636482 TI - Evidence for dynamic heterogeneities in computer simulations of miscible polymer blends. AB - The controversial origins of the unusual dynamics of miscible polymer blends are incisively probed through computer simulations. The distribution of mobilities experienced by a probe monomer in a miscible blend of chains with disparate glass transition temperatures is found to be much broader than in the pure polymers, providing clear evidence for local concentration variations in the mixture. These concentration fluctuations yield distinctly different temperature dependences for the dynamics of the two different components, in a manner that closely mimics experiments. PMID- 12636481 TI - Nematics with dispersed polymer fibrils: a Monte Carlo study of the external field-induced switching. AB - We present a Monte Carlo study of molecular ordering in nematics with dispersed regular and random arrays of straight and distorted polymer fibrils. We focus on the collective molecular reorientation--the switching--resulting from the competing aligning effects of fibrils and of a progressively applied transversal external field, and for straight fibrils identify structural Freedericksz and saturation transitions. The role of fibril topography in the switching is monitored by simulating electric capacitance Slightly distorted fibrils are shown to give a sharper switching at a lower threshold. PMID- 12636483 TI - Local structure and glass transition of polybutadiene up to 4 GPa. AB - This communication presents a determination of the glass transition of polybutadiene under very high pressure, and raises the problem of the determination of the relative effects of temperature and density on the glass transition, depending on the pressure and temperature conditions. Local structure and slow dynamics were studied, by neutron scattering and calorimetry. To the best of our knowledge in neutron diffraction on soft matter such a high pressure, up to 4 GPa, was achieved. PMID- 12636485 TI - Equivalence of driven and aging fluctuation-dissipation relations in the trap model. AB - We study the nonequilibrium version of the fluctuation-dissipation (FD) relation in the glass phase of a trap model that is driven into a nonequilibrium steady state by external "shear." This extends our recent study of aging FD relations in the same model, where we found limiting, observable independent FD relations for "neutral" observables that are uncorrelated with the system's average energy. In this work, for such neutral observables, we find the FD relation for a stationary weakly driven system to be the same, to within small corrections, as for an infinitely aged system. We analyze the robustness of this correspondence with respect to non-neutrality of the observable, and with respect to changes in the driving mechanism. PMID- 12636484 TI - Testing the null hypothesis of the nonexistence of a preseizure state. AB - A rapidly growing number of studies deals with the prediction of epileptic seizures. For this purpose, various techniques derived from linear and nonlinear time series analysis have been applied to the electroencephalogram of epilepsy patients. In none of these works, however, the performance of the seizure prediction statistics is tested against a null hypothesis, an otherwise ubiquitous concept in science. In consequence, the evaluation of the reported performance values is problematic. Here, we propose the technique of seizure time surrogates based on a Monte Carlo simulation to remedy this deficit. PMID- 12636486 TI - Mechanical model of normal and anomalous diffusion. AB - The overdamped dynamics of a charged particle driven by an uniform electric field through a random sequence of scatterers in one dimension is investigated. Analytic expressions of the mean velocity and of the velocity power spectrum are presented. These show that above a threshold value of the field normal diffusion is superimposed to ballistic motion. The diffusion constant can be given explicitly. At the threshold field, the transition between conduction and localization is accompanied by an anomalous diffusion. Our results exemplify that, even in the absence of time-dependent stochastic forces, a purely mechanical model equipped with a quenched disorder can exhibit normal as well as anomalous diffusion, the latter emerging as a critical property. Via another interpretation, as the motion of a particle on an inclined rough surface, our results are relevant for the problem of segregation by flow. PMID- 12636487 TI - Intramolecular coupling as a mechanism for a liquid-liquid phase transition. AB - We study a model for water with a tunable intramolecular interaction J(sigma), using mean-field theory and off-lattice Monte Carlo simulations. For all J(sigma)> or =0, the model displays a temperature of maximum density. For a finite intramolecular interaction J(sigma)>0, our calculations support the presence of a liquid-liquid phase transition with a possible liquid-liquid critical point for water, likely preempted by inevitable freezing. For J=0, the liquid-liquid critical point disappears at T=0. PMID- 12636488 TI - Thermal decay of a metastable state: exact solution. AB - Exact solutions are obtained for the thermal decay of a metastable state for two different forms of the potential and for uniform and localized boundary conditions by using the Laplace transform method. The exact inverse Laplace transforms are found for a symmetric case, and the results are compared with other calculations and with the Kramers rate. PMID- 12636489 TI - Dynamical origin of uniform sampling in multicanonical ensemble. AB - The stochastic model describing the sampling process in multicanonical ensemble has been derived by considering the sampling process as an overdamped Brownian motion on the free energy surface. The essential dynamics of the multicanonical sampling has been characterized by a Langevin equation in a piecewise multivalleyed free energy landscape, modulated by a temperature-dependent curvature. Based on the stochastic model we showed that the multicanonical weight can be determined by interpolating maximum probability energy points of the canonical samplings at different temperatures. PMID- 12636490 TI - Combined effect of periodic gates and external fields on the diffusion coefficient of a single particle. AB - A general analytical expression to describe the diffusion of a single particle in a one-dimensional lattice with periodically distributed gates of lifetime (tau) and while under the influence of a constant external field is calculated. A formulation based on a microscopic model and a diffusion relaxation condition is used to derive an equation for the diffusion coefficient as a function of the concentration of gates (c), the lifetime (tau) of such gates, and the strength of the external field (p). The theory is compared against Monte Carlo simulations, and limiting cases are used to reproduce previously published results on a variety of phenomena. PMID- 12636491 TI - Temperature of maximum density line of a polarizable water model. AB - Monte Carlo simulations of liquid water were performed with a polarizable model in a narrow range of temperatures around the temperature of maximum density (TMD) and a broad pressure range, extended also to negative pressures. In order to infer features of the phase diagram relevant for the explanation of the anomalous behavior of water the pressure dependence of the TMD line was analyzed. The TMD is found to increase with decreasing pressure up to 274.8 K at -32 bar, and decrease upon further stretching. This behavior excludes the possibility of a reentrant liquid-gas spinodal line. PMID- 12636492 TI - Analytic study of the urn model for separation of sand. AB - We present an analytic study of the urn model for separation of sand recently introduced by Lipowski and Droz [Phys. Rev. E 65, 031307 (2002)]. We solve analytically the master equation and the first-passage problem. The analytic results confirm the numerical results obtained by Lipowski and Droz. We find that the stationary probability distribution and the shortest one among the characteristic times are governed by the same free energy. We also analytically derive the form of the critical probability distribution on the critical line, which supports their results obtained by numerically calculating Binder cumulants (A. Lipowski and M. Droz, e-print cond-mat/0201472). PMID- 12636493 TI - Instabilities in slowly driven granular packing. AB - In this work, a digital imaging technique is used to study the superficial fluctuations observed when a granular packing is slowly driven to the threshold of instability. The experimental results show the presence of three types of events. Small superficial rearrangements of grains are observed during all the experiments. They present a power-law behavior although the system is not in a critical state as predicted by self-organized criticality models. In thick granular piles, large rearrangements are detected at regular angular intervals. They are related to the threshold of instability of the contact network that relaxes to stable configurations producing internal rearrangements of the grains. Finally, an avalanche is triggered when the superficial beads that are set in motion acquire enough momentum to destabilize grains from layers below. PMID- 12636494 TI - Rolling motion of a bead in a rapid water stream. AB - This paper investigates the two-dimensional rolling motion of a single large particle in a shallow water stream down a steep rough bed from both an experimental and a theoretical point of view. The experiment is prototypal of sediment transport on sloping beds. Two theoretical models are presented. The first model uses the mean kinetic energy balance to deduce the average particle velocity and the bounds of the flow-rate range within which a rolling regime occurs. This range is found to be narrow, which means that the fully rolling regime is a marginal mode of transport between repose and saltation. In the second model, the particle state (resting, rolling, saltating) is considered as a random variable, whose evolution constitutes a jump Markov chain. This makes it possible to deduce the mean particle velocity as a function of the flow conditions without explicit mention of its state. The theoretical results are finally compared to the experimental data. The second model provides correct estimates of the particle velocity and the probability of finding the particle in a given state for various flow conditions (bead material, slope, and roughness). PMID- 12636495 TI - Measurements of particle dynamics in slow, dense granular Couette flow. AB - Experimental measurements of particle dynamics on the lower surface of a three dimensional (3D) Couette cell containing monodisperse spheres are reported. The average radial density and velocity profiles are similar to those previously measured within the bulk and on the lower surface of the 3D cell filled with mustard seeds. Observations of the evolution of particle velocities over time reveal distinct motion events, intervals where previously stationary particles move for a short duration before jamming again. The cross correlation between the velocities of two particles at a given distance r from the moving wall reveals a characteristic length scale over which the particles are correlated. The autocorrelation of a single particle's velocity reveals a characteristic time scale tau, which decreases with increasing distance from the inner moving wall. This may be attributed to the increasing rarity at which the discrete motion events occur and the reduced duration of those events at large r. The relationship between the rms azimuthal velocity fluctuations, delta v(theta)(r), and average shear rate, gamma(r), was found to be delta v(theta) is proportional to gamma(alpha) with alpha=0.52+/-0.04. These observations are compared with other recent experiments and with the modified hydrodynamic model recently introduced by Bocquet et al. PMID- 12636496 TI - Model for gelation with explicit solvent effects: structure and dynamics. AB - We study a two-component model for gelation consisting of f-functional monomers (the gel) and inert particles (the solvent). After equilibration as a simple liquid, the gel particles are gradually cross linked to each other until the desired number of cross links have been attained. At a critical cross-link density, the largest gel cluster percolates and an amorphous solid forms. This percolation process is different from ordinary lattice or continuum percolation of a single species in the sense that the critical exponents are new. As the cross-link density p approaches its critical value p(c), the shear viscosity diverges: eta(p) approximately (p(c)-p)(-s) with s a nonuniversal concentration dependent exponent. PMID- 12636497 TI - Clustering under short-range finite interactions. AB - In this paper the aggregation of surface modified colloidal particles is presented, paying special attention to the cluster structure and growth. The surface was modified by adsorbing bovine serum albumin (BSA). The interaction potential develops a minimum of restricted depth, weakening the clusters which subsequently restructure and form more compact morphologies. This minimum is responsible for the reversibility of the aggregation processes (this is an important difference between diffusion-limited cluster aggregation and reaction limited cluster aggregation). The energy minimum is associated with the presence of a steric term in the energy balance, which depends on the size of the adsorbed molecules. BSA molecules with different sizes were employed to test this point. In addition, the short-range interaction seems not to affect significantly the paths of approximating particles, since the aggregation of the clusters at long times is independent of the size of these particles. The long-time kinetics was interpreted in the frame of dynamic scaling concepts. A kinetics model, including surface-surface, protein-surface, and protein-protein aggregation, is used to determine the dominant mechanism controlling the aggregation. PMID- 12636498 TI - Crystal phases of a glass-forming Lennard-Jones mixture. AB - We compare the potential energy at zero temperature of a range of crystal structures for a glass-forming binary mixture of Lennard-Jones particles. The lowest-energy ordered state consists of coexisting phases of a single component face centered cubic structure and an equimolar cesium chloride structure. An infinite number of layered crystal structures are identified with energies close to this ground state. We demonstrate that the finite size increase of the energy of the coexisting crystal with incoherent interfaces is sufficient to destabilize this ordered phase in simulations of typical size. Two specific local coordination structures are identified as of possible structural significance in the amorphous state. We observe rapid crystal growth in the equimolar mixture. PMID- 12636499 TI - Colloid transport in nonuniform temperature. AB - The aim of this paper is to set up a theoretical framework for obtaining the thermodiffusion (or Soret) coefficient of a colloid in a carrier liquid. It is first argued that the expression of the particle-current density in nonuniform temperature cannot be derived from a theoretical formula valid for an isothermal solution. Then the kinetic theory of Brownian motion is used to derive an expression for the current density properly accounting for thermodiffusion. The cases of free and interacting particles are treated, and the thermodiffusion current pertinent to an ideal solution adds up with a current driven by a temperature- and concentration-dependent potential. Accordingly, a general explicit formula for the thermodiffusion coefficient is obtained. Practical use of the framework is illustrated on simple specific models of a colloid in a solvent. Large Soret coefficients of both signs are calculated for realistic values of the physicochemical parameters, in qualitative agreement with published experimental data. PMID- 12636500 TI - Dipole fluid as a basic model for the equation of state of ionic liquids in the vicinity of their critical point. AB - A model of dipole fluid for ionic liquids similar to molten NaCl is proposed. The estimates for the critical parameters are obtained with the help of the van der Waals equation of state. The influence of the rotation on the characteristics of a dipole pair and the location of the critical point is discussed. The dissociation of such a fluid near the critical point is considered. PMID- 12636501 TI - Extensional rupture of model non-Newtonian fluid filaments. AB - We present molecular dynamics computer simulations of filaments of a model non Newtonian liquid stretched in a uniaxial deformation to the point of breaking. The liquid consists of Lennard-Jones monomers bound into chains of 100 monomers by nonlinear springs, and several different constant-velocity and constant-strain rate deformations are considered. Generally we observe nonuniform extensions originating in an interplay between the stretching forces and elastic and capillary restoring mechanisms, leading to highly uneven shapes and alternating stretched and unstretched regions of liquid. Except at the fastest pulling speeds, the filaments continue to thin indefinitely and break only when depleted of molecules, rather than due to common viscoelastic rupture mechanisms. PMID- 12636502 TI - Phase diagrams of hard spheres with algebraic attractive interactions. AB - The phase diagrams of systems made up of hard spheres interacting with attractive potentials of the form -1/r(3+sigma) are calculated using Monte Carlo simulations, second-order thermodynamic perturbation theory, and an augmented van der Waals theory. In simulations of the systems with sigma=0.1, 1, and 3, fluid solid coexistence results are obtained using the Gibbs-Duhem integration technique; simulation data for the vapor-liquid coexistence envelopes and critical points are taken from previously published work [P. J. Camp and G. N. Patey, J. Chem. Phys. 114, 399 (2001)]. It is shown that the agreement between the theoretical and simulated phase diagrams improves as the range of the potential is increased, reflecting the decreasing role of short-range correlations in determining the bulk thermodynamics. In the extreme case of sigma=0.1 both theories are in excellent agreement with simulations. Phase diagrams for systems with sigma=4, 5, and 6 are computed using second-order thermodynamic perturbation theory. The results indicate that the vapor-liquid transition becomes metastable with respect to freezing when sigma > or approximately equal to 5, in broad agreement with results for the hard-sphere attractive Yukawa system which is commonly used to model colloidal particles, globular proteins, and nanoparticles. PMID- 12636503 TI - Rotational magnetic particles microrheology: the Maxwellian case. AB - An experimental method based on the rotational dynamics of a magnetic probe is reported to measure the local viscoelasticity of soft materials on microscopic scales. The technique is based on the alignment of dipolar chains of submicrometer magnetic particles in the direction of an applied magnetic field. On one hand, light scattering is used to detect the chains' oscillations over a 0.001-100 Hz frequency range when submitted to an oscillating magnetic field and leads to global microrheological measurements. On the other hand, the chains' rotation toward a permanent magnetic field is observed with a microscope, allowing a local determination of viscoelastic properties on the scale of the chains of particles. We demonstrate the accuracy of both assays with a micellar Maxwellian solution and validate theoretical predictions. PMID- 12636504 TI - Improved simulation method for the calculation of the elastic constants of crystalline and amorphous systems using strain fluctuations. AB - In this paper, a method is proposed for calculating the elastic constants of arbitrarily soft or stiff systems using strain fluctuations. For stiff materials, for example, strain fluctuations may be enhanced by appropriate choice of elastic constants for the bath. Example calculations of the isothermal elastic constants of the nearest-neighbor Lennard-Jones fcc crystal demonstrate improved convergence properties over standard techniques. Elastic constants for a model amorphous polymer system are also presented. PMID- 12636505 TI - Ground state of a polydisperse electrorheological solid: beyond the dipole approximation. AB - The ground state of an electrorheological (ER) fluid has been studied based on our recently proposed dipole-induced dipole (DID) model. We obtained an analytical expression of the interaction between chains of particles which are of the same or different dielectric constants. The effects of dielectric constants on the structure formation in monodisperse and polydisperse electrorheological fluids are studied in a wide range of dielectric contrasts between the particles and the base fluid. Our results showed that the established body-centered tetragonal ground state in monodisperse ER fluids may become unstable due to a polydispersity in the particle dielectric constants. While our results agree with that of the fully multipole theory, the DID model is much simpler, which offers a basis for computer simulations in polydisperse ER fluids. PMID- 12636506 TI - Scale invariance in coarsening of binary and ternary fluids. AB - Phase separation in binary and ternary fluids is studied using a two-dimensional lattice gas automata. The lengths given by the the first zero crossing point of the correlation function and the total interface length is shown to exhibit power law dependence on time. In binary mixtures, our data clearly indicate the existence of a regime having more than one length scale, where the coarsening process proceeds through the rupture and reassociation of domains. In ternary fluids; in the case of symmetric mixtures there exists a regime with a single length scale having dynamic exponent 1/2, while in asymmetric mixtures our data establish the break down of scale invariance. PMID- 12636507 TI - Spinodal decomposition, power laws, and wetting at a triple point. AB - We study numerically the dynamics of phase separation in ternary mixtures at a triple point. For the full range of compositions and for different interaction parameters, the long time growth is in accord with a universal law. The early time behavior is governed by the structure of the spinodal region, including the possibility of a two step separation and decomposition originating at a surface and propagating into the bulk. The appearance of the domains is governed by the wetting properties of the mixture and the growth of a wetting layer follows again the universal law; a result that we can interpret with a simple phenomenological model. PMID- 12636508 TI - Asymptotic function for multigrowth surfaces using power-law noise. AB - Numerical simulations are used to investigate the multiaffine exponent alpha(q) and multigrowth exponent beta(q) of ballistic deposition growth for noise obeying a power-law distribution. The simulated values of beta(q) are compared with the asymptotic function beta(q)=1/q that is approximated from the power-law behavior of the distribution of height differences over time. They are in good agreement for large q. The simulated alpha(q) is found in the range 1/q< or =alpha(q)< or =2/(q+1). This implies that large rare events tend to break the Kardar-Parisi Zhang universality scaling law at higher order q. PMID- 12636509 TI - Epitaxial growth with pulsed deposition: submonolayer scaling and Villain instability. AB - It has been observed experimentally that under certain conditions, pulsed laser deposition (PLD) produces smoother surfaces than ordinary molecular beam epitaxy (MBE). So far, the mechanism leading to the improved quality of surfaces in PLD is not yet fully understood. In the present work, we investigate the physical properties of a simple model for PLD, in which the transient mobility of adatoms and diffusion along edges is neglected. Analyzing the crossover from MBE to PLD, the scaling properties of the time-dependent nucleation density as well as the influence of Ehrlich-Schwoebel barriers, we find that there is indeed a range of parameters, where the surface quality in PLD is better than in MBE. However, since the improvement is weak and occurs only in a small range of parameters we conclude that deposition in pulses alone cannot explain the experimentally observed smoothness of PLD-grown surfaces. PMID- 12636510 TI - Molecular simulations of liquid-liquid interfacial properties: water-n-alkane and water-methanol-n-alkane systems. AB - Direct molecular dynamics simulations of the liquid-liquid interface of water-n alkane and water-methanol-n-alkane systems have been performed in order to study the interfacial properties of these systems. The simulations were carried out using the NERD revised force field of Nath et al. for the n-alkanes, the simple point charge extended model for water, and the optimized potential for liquid simulations model for methanol. In order to validate the model employed in this work for the n-alkanes we calculated the coexisting densities, surface tension, and thickness of the interface for pure n-pentane. For all the systems studied the interfacial tension and thickness were calculated at 298.15 K. Our results show that, by adjusting the number of molecules to reproduce the liquid densities in the direct simulation method of the liquid-liquid interface in multicomponent systems, we are able to reproduce available experimental data for interfacial tension. The interfacial thickness is underpredicted and a constant negative deviation of approximately 2.5 A from the experimental data is usually observed. We find that methanol acts like surfactant when it is added to the water-n-alkane mixtures, reducing the interfacial tension of the liquid-liquid ternary system. The interfacial tension results agree quantitatively well for the range of concentrations of methanol studied. PMID- 12636511 TI - Isotropic-to-cholesteric transition in liquid crystal elastomers. AB - A liquid crystal elastomer tries to spontaneously elongate on entering the locally nematic phase, but may have to twist to reduce its Frank elastic energy. The extremes are a conventional, transverse cholesteric structure (where it reduces its Frank energy), and a uniformly aligned state (where it can maximally spontaneously extend and reduce its elastic energy). Between these it can adopt a conical state where there is also bend but equally a partial satisfaction of the elastic requirements. A line of first-order transitions between conical and transverse states ends and becomes a line of second-order transitions, depending on chain anisotropy, the ratio of the Frank bend and twist constants, and on the elastic modulus reduced by the bend energy. Continuous and discontinuous variation of cone angles, and spontaneous elongations and shears are given, as are analytic forms for the singular variation of director as cones are lost to the transverse state. The variation of the multicritical point with the ratio of Frank constants is also given. PMID- 12636512 TI - Spherical particle immersed in a nematic liquid crystal: effects of confinement on the director field configurations. AB - The effects of confinement on the director field configurations are studied for a spherical particle immersed in a nematic liquid crystal. The liquid crystal is confined in a cylindrical geometry and the particle is located on the axis of symmetry. A finite element method is used to minimize the Frank free energy for various sizes of the system. The liquid crystal is assumed to possess strong anchoring at all the surfaces in the system. Two structures are examined for strong homeotropic anchoring at the surface of the particle: configuration with a Saturn ring disclination line and configuration with a satellite point defect (hedgehog defect). It is shown that the equilibrium locations of the Saturn ring and of the hedgehog point defect change with confinement. It is also found that confinement induces an increase in the elastic free energy that differs substantially with the type of topological defect under consideration. In particular, the evaluation of the total free energy that includes an approximate contribution for the core defect shows that, for micrometer-sized particles in confined systems, the Saturn ring configuration appears to be more stable than the hedgehog defect. This result is in contrast to the bulk situation, where the hedgehog is more stable than the Saturn ring, and it helps explain recent experimental observations of Saturn ring defects around confined micrometer-sized solid particles. PMID- 12636513 TI - Phase behavior of bent-core molecules. AB - Recently, a new class of smectic liquid crystal phases characterized by the spontaneous formation of macroscopic chiral domains from achiral bent-core molecules has been discovered. We have carried out Monte Carlo simulations of a minimal hard spherocylinder dimer model to investigate the role of excluded volume interactions in determining the phase behavior of bent-core materials and to probe the molecular origins of polar and chiral symmetry breaking. We present the phase diagram of hard spherocylinder dimers of length-diameter ratio of 5 as a function of pressure or density and dimer opening angle psi. With decreasing psi, a transition from a nonpolar to a polar smectic A phase is observed near psi=167 degrees, and the nematic phase becomes thermodynamically unstable for psi<135 degrees. Free energy calculations indicate that the antipolar smectic A (SmAP(A)) phase is more stable than the polar smectic A phase (SmAP(F)). No chiral smectic or biaxial nematic phases were found. PMID- 12636514 TI - Apparent tricritical behavior at a nearly second-order nematic-isotropic phase transition of a cyclic liquid crystalline trimer. AB - The cyclic liquid crystalline trimer TPB-(c)9(3) was investigated by optical retardation and Freedericksz techniques within a few tens of millikelvins of the superheating limit of the nearly second-order nematic-isotropic phase transition. Both the optical retardation and the Freedericksz bend threshold voltage are in good agreement with tricritical behavior for the transition. PMID- 12636515 TI - Monte Carlo simulation of a planar lattice model with P4 interactions. AB - Monte Carlo study of a two-dimensional lattice with three-dimensional spins (d=2,n=3) interacting with nearest neighbors via a -P4(cos theta) potential, where P4 is the fourth Legendre polynomial and theta is the angle between two spins, has been reported for lattice sizes ranging from 10 x 10 to 160 x 160. A cluster algorithm for spin updating with a histogram reweighting technique has been used and finite size scaling has been performed. The model exhibits a strong first order phase transition at a dimensionless temperature 0.376+/-0.015. The phase transition appears to be driven by condensation of topological defects and the defect density D increases sharply at the transition temperature. The temperature derivative dD/dT* is found to obey a linear scaling relation with the lattice size L. The behavior of the model seems to be remarkably different from the two-dimensional P2 model, that has been investigated by other authors, although both models possess the same symmetry and topological defects play an important role in the phase transition. PMID- 12636516 TI - Nematostatics of triple lines. AB - The Landau-de Gennes model for nematic liquid crystal bulk and interfaces has been extended to nematic triple lines involving the intersection of two isotropic fluids and one nematic liquid crystalline phase. A complete set of bulk, interface, and triple line force and torque balance equations has been formulated. The triple line force and torque balance equations have linear, interfacial, and bulk contributions. The bulk contributions appear as junction integrals, the surface contributions as junctions sums, and the line contributions as gradients of stresses. Reduction of dimensionality from three to one dimensional creates the following effects: (a) bulk terms enter interfacial balances as surface jumps and line balances as junction integrals, and (b) surface terms enter linear balances as junction sums. Line stress and torque equations are derived using classical liquid crystal models. The correspondence between line stress and line torque and their surface and bulk analogs is established. The triple line force and torque balance equations are use to analyze the contact angle in a nematic lens lying at the interface between two isotropic fluids, when the preferred surface orientation is tangential. The effect of anisotropy and long range elasticity on triple line phases is established. Under weak anchoring the contact angle is shown to be a function of the anchoring energy at the nematic-isotropic interface, while under strong anchoring conditions the contact angle is a function of the Peach-Koehler force that originates from bulk long range elasticity and acts on the triple line. The use of the complete set of balance equations removes the classical inconsistency in force balances at a contact line by properly taking into account long range (bulk gradient elasticity) and anisotropic (interfacial anchoring elasticity) effects. PMID- 12636517 TI - Columnar phases of three-fold molecular structures. AB - We study the occurrence of both positional and helical orders, for three-fold symmetric flat molecules in a columnar liquid crystal phase D(hd). Working in the group-theoretical Landau's framework, we identify a set of three order parameters giving rise to supercells of intercalated column with helical order and opposite helicities, as observed experimentally. The first-order nature of the transition is also discussed. The degrees of freedom introduced give rise to various other phases, including a new type, having helical order but no positional order along the columns. PMID- 12636518 TI - Smectic ordering in liquid-crystal-aerosil dispersions. I. X-ray scattering. AB - Comprehensive x-ray scattering studies have characterized the smectic ordering of octylcyanobiphenyl (8CB) confined in the hydrogen-bonded silica gels formed by aerosil dispersions. For all densities of aerosil and all measurement temperatures, the correlations remain short range, demonstrating that the disorder imposed by the gels destroys the nematic (N) to smectic-A (SmA) transition. The smectic correlation function contains two distinct contributions. The first has a form identical to that describing the critical thermal fluctuations in pure 8CB near the N-SmA transition, and this term displays a temperature dependence at high temperatures similar to that of the pure liquid crystal. The second term, which is negligible at high temperatures but dominates at low temperatures, has a shape given by the thermal term squared and describes the static fluctuations due to random fields induced by confinement in the gel. The correlation lengths appearing in the thermal and disorder terms are the same and show a strong variation with gel density at low temperatures. The temperature dependence of the amplitude of the static fluctuations further suggests that nematic susceptibility becomes suppressed with increasing quenched disorder. The results overall are well described by a mapping of the liquid-crystal-aerosil system onto a three-dimensional XY model in a random field with disorder strength varying linearly with the aerosil density. PMID- 12636519 TI - Smectic ordering in liquid-crystal-aerosil dispersions. II. Scaling analysis. AB - Liquid crystals offer many unique opportunities to study various phase transitions with continuous symmetry in the presence of quenched random disorder (QRD). The QRD arises from the presence of porous solids in the form of a random gel network. Experimental and theoretical work supports the view that for fixed (static) inclusions, quasi-long-range smectic order is destroyed for arbitrarily small volume fractions of the solid. However, the presence of porous solids indicates that finite-size effects could play some role in limiting long-range order. In an earlier work, the nematic-smectic-A transition region of octylcyanobiphenyl (8CB) and silica aerosils was investigated calorimetrically. A detailed x-ray study of this system is presented in the preceding paper, which indicates that pseudocritical scaling behavior is observed. In the present paper, the role of finite-size scaling and two-scale universality aspects of the 8CB+aerosil system are presented and the dependence of the QRD strength on the aerosil density is discussed. PMID- 12636520 TI - Switching dynamics and surface forces in thresholdless "V-shaped" switching ferroelectric liquid crystals. AB - The electrostatic model of thresholdless V-shaped switching is able to explain the general low frequency (quasistatic) electro-optic behavior of smectic-C* ferroelectric liquid crystals (vFLCs). Here, dynamical equations based on the electrostatic model are developed which predict a vFLC cell's small-amplitude switching speed and which also show that a strongly amplitude-dependent switching speed is expected. A relationship between the switching time constants of analog vFLCs and of the faster, yet structurally similar, binary FLCs is found. The electrostatic model applies in the limit where the FLC's spontaneous polarization is large enough to completely overwhelm surface and elastic forces. This analysis suggests that, in many cases of practical interest, electrostatic energies may be low enough for surface forces to play an important role even when the director structure is strongly stiffened by a large polarization charge. It is shown that the addition of surface forces to the electrostatic model can improve agreement between the model and the observed dynamical response of vFLC cells. PMID- 12636521 TI - Stiff polymer in monomer ensemble. AB - We employ an ordered monomer ensemble formalism in order to develop techniques to investigate a stiff polymer chain which is confined to a certain region. In particular, we calculate the segment density for a given location and segment orientation distribution within the confining geometry. With this method the role of the stiffness can be examined by means of differential equations, integral equations, or recursive relations for both continuum and lattice models. A suitable choice of lattice model permits an exact analytical solution for the segment location and orientation density for a chain between two parallel plates. For the stiff polymer in a spherical cavity we develop an integral equation formalism which is treated numerically, and in the same spherical geometry, a different model of the polymer displays a solution of a differential equation. PMID- 12636522 TI - Interacting growth walk: a model for hyperquenched homopolymer glass? AB - We show that the compact self-avoiding walk configurations, kinetically generated by the recently introduced interacting growth walk (IGW) model, can be considered as members of a canonical ensemble if they are assigned random values of energy. Such a mapping is necessary for studying the thermodynamic behavior of this system. We have presented the specific heat data for the IGW, obtained from extensive simulations on a square lattice; we observe a broad hump in the specific heat above the theta point, contrary to expectation. PMID- 12636523 TI - Self-consistent dynamics of wall slip. AB - A simple molecular model is studied to explain wall slip in a polymer melt. We consider a tube model for tethered chains in which the most important relaxation mechanisms: convective constraint release and chain stretching (retraction), are incorporated. Furthermore, we take the interactions between tethered chains and bulk flow self-consistently into account. Numerical simulations show that our model exhibits an entanglement-disentanglement transition, leading to a jump in the slip velocity which increases with the number of entanglements and the grafting density. The wall shear stress is found to be a nonmonotonic function of the slip and plate velocity, yielding the possibility of hysteresis and spurt instabilities. In a simplified version of the model we show via an analytical approach that the stick-slip transition is asymmetrical: the transition from stick to slip is much faster than the slip to stick transition. Our analysis reveals the existence of a dimensionless parameter that determines the time scale of the dynamics for the slowing down of the bulk flow. The relative rate at which relaxation of the tethered chains and slowing down of the bulk take place, seems to be quintessential for the slip behavior of the melt. PMID- 12636524 TI - Counterion penetration and effective electrostatic interactions in solutions of polyelectrolyte stars and microgels. AB - Counterion distributions and effective electrostatic interactions between spherical macroions in polyelectrolyte solutions are calculated via second-order perturbation (linear response) theory. By modeling the macroions as continuous charge distributions that are permeable to counterions, analytical expressions are obtained for counterion profiles and effective pair interactions in solutions of star-branched and microgel macroions. The counterions are found to penetrate stars more easily than microgels, with important implications for screening of bare macroion interactions. The effective pair interactions are Yukawa in form for separated macroions, but are softly repulsive and bounded for overlapping macroions. A one-body volume energy, which depends on the average macroion concentration, emerges naturally in the theory and contributes to the total free energy. PMID- 12636525 TI - Persistence length of a strongly charged rodlike polyelectrolyte in the presence of salt. AB - The persistence length of a single, intrinsically rigid polyelectrolyte chain, above the Manning condensation threshold is investigated theoretically in the presence of added salt. Using a loop expansion method, the partition function is consistently calculated, taking into account corrections to mean-field theory. Within a mean-field approximation, the well-known results of Odijk, Skolnick, and Fixman are reproduced. Beyond mean field, it is found that density correlations between counterions and thermal fluctuations reduce the stiffness of the chain, indicating an effective attraction between monomers for highly charged chains and multivalent counterions. This attraction results in a possible mechanical instability (collapse), alluding to the phenomenon of DNA condensation. In addition, we find that more counterions condense on slightly bent conformations of the chain than predicted by the Manning model for the case of an infinite cylinder. Finally, our results are compared with previous models and experiments. PMID- 12636526 TI - Relationship between the nonlinear ferroelectric and liquid crystal models for microtubules. AB - Microtubules (MTs), which are the main components of the cytoskeleton, are important in a variety of cellular activities, but some physical properties underlying the most important features of their behavior are still lacking satisfactory explanation. One of the essential enigmas regarding the energy balance in MTs is the hydrolysis of the exchangeable guanosine 5'-triphosphate bound to the beta monomer of the molecule. The energy released in the hydrolysis process amounts to 6.25 x 10(-20) J and has been the subject of many attempts to answer the questions of its utilization. Earlier, we put forward a hypothesis that this energy can cause a local conformational distortion of the dimer. This distortion should have nonlinear character and could lead to the formation of a traveling kink soliton. In this paper we use the formalism of the liquid crystal theory to consider the nonlinear dynamics of MTs. We demonstrate that this new model is formally equivalent to our earlier ferroelectric model which was widely exploited in an attempt to elucidate some important dynamical activities in MTs. We also study the stability of kink solitons against small perturbations and their unusual mutual interactions as well as the interactions with structural inhomogeneities of MTs. Our new approach based on liquid crystal properties of microtubules has been recently corroborated by new insights gained from the electrostatic properties of tubulin and microtubules. PMID- 12636527 TI - Exponential stability and periodic solutions of neural networks with continuously distributed delays. AB - In this paper we study a class of neural networks with continuously distributed delays. By means the of Lyapunov functional method, we obtain some sufficient conditions ensuring the existence, uniqueness, and global exponential stability of the equilibrium and periodic solution. We also estimate the exponentially convergent rate. Our results are less restrictive than previously known criteria and can be applied to neural networks with a broad range of activation functions assuming neither differentiability nor strict monotonicity. Moreover, these conclusions are presented in terms of system parameters and can be easily verified. Therefore, our results play an important role in the design of globally exponentially stable neural circuits and periodic oscillatory neural circuits. PMID- 12636528 TI - Networks with fourfold connectivity in two dimensions. AB - The elastic properties of planar, C4-symmetric networks under stress and at nonzero temperature are determined by simulation and mean field approximations. Attached at fourfold coordinated junction vertices, the networks are self avoiding in that their elements (or bonds) may not intersect each other. Two different models are considered for the potential energy of the elements: either Hooke's law springs or flexible tethers (square well potential). For certain ranges of stress and temperature, the properties of the networks are captured by one of several models: at large tensions, the networks behave like a uniform system of square plaquettes, while at large compressions or high temperatures, they display many characteristics of an ideal gas. Under less severe conditions, mean field models with more general shapes (parallelograms) reproduce many essential features of both networks. Lastly, the spring network expands without limit at a two-dimensional tension equal to the force constant of the spring; however, it does not appear to collapse under compression, except at zero temperature. PMID- 12636529 TI - Structure of self-assembled liposome-DNA-metal complexes. AB - We have studied the structural and morphological properties of the triple complex dioleoyl phosphatidylcholine (DOPC)-DNA-Mn2+ by means of synchrotron x-ray diffraction and freeze-fracture transmission electron microscopy. This complex is formed in a self-assembled manner when water solutions of neutral lipid, DNA, and metal ions are mixed, which represents a striking example of supramolecular chemistry. The DNA condensation in the complex is promoted by the metal cations that bind the polar heads of the lipid with the negatively charged phosphate groups of DNA. The complex is rather heterogeneous with respect to size and shape and exhibits the lamellar symmetry of the L(c)(alpha) phase: the structure consists of an ordered multilamellar assembly similar to that recently found in cationic liposome-DNA complexes, where the hydrated DNA helices are sandwiched between the liposome bilayers. The experimental results show that, at equilibrium, globules of the triple complex in the L(c)(alpha) phase coexist with globules of multilamellar vesicles of DOPC in the L(alpha) phase, the volume ratio of the two structures being dependent on the molar ratio of the three components DOPC, DNA, and Mn2+. These complexes are of potential interest for applications as synthetically based nonviral carriers of DNA vectors for gene therapy. PMID- 12636530 TI - Dynamics-based sequential memory: winnerless competition of patterns. AB - We introduce a biologically motivated dynamical principle of sequential memory which is based on winnerless competition (WLC) of event images. This mechanism is implemented in a two-layer neural model of sequential spatial memory. We present the learning dynamics which leads to the formation of a WLC network. After learning, the system is capable of associative retrieval of prerecorded sequences of patterns. PMID- 12636531 TI - Dynamical and stationary properties of on-line learning from finite training sets. AB - The dynamical and stationary properties of on-line learning from finite training sets are analyzed by using the cavity method. For large input dimensions, we derive equations for the macroscopic parameters, namely, the student-teacher correlation, the student-student autocorrelation and the learning force fluctuation. This enables us to provide analytical solutions to Adaline learning as a benchmark. Theoretical predictions of training errors in transient and stationary states are obtained by a Monte Carlo sampling procedure. Generalization and training errors are found to agree with simulations. The physical origin of the critical learning rate is presented. Comparison with batch learning is discussed throughout the paper. PMID- 12636532 TI - Why biomolecules prefer only a few crystal structures. AB - We have shown that, in determining the biomolecule-crystal symmetry, the occupation of low-site-symmetry Wyckoff positions is crucial, which contrasts with the overwhelming majority of nonmolecular, inorganic crystals where atoms mainly reside in high-symmetry Wyckoff positions. We consider the general relation between the symmetry of an isolated molecule and the possible symmetries of biomolecular crystals it can generate. We reveal that the improper symmetry operations (inversion and mirror symmetries) are prohibited in the chirally pure biomolecular crystals. Next, we show that the low (C1) symmetry of large biological molecules substantially decreases the space in a crystal where the molecules can reside. The space "forbidden" for molecule centers is found to be in the R vicinity of the higher-symmetry Wyckoff positions on symmetry lines, where R is the molecule characteristic size. The remaining free space and hence the probability for the structure to exist are shown to be drastically increased when replacing any rotation axis by a screw one. Based on the proposed model, we have explained the peculiar distribution of biomolecular crystals over the space groups, which can be obtained from biomolecule-crystal databases. PMID- 12636533 TI - Protein-induced morphological transitions in KCl crystal growth. AB - We investigated the formation of KCl crystals on a glass surface by phase contrast, fluorescent, and atomic force microscopy on the micrometer scale and observed interesting morphological transitions as a function of the experimental conditions. The presence of proteins in the solution from which the salt crystals grow during the drying up leads to complex microscopic patterns of crystals, some of which are analogous to those commonly observed on the macroscopic scale. We tested the effect of tubulin, FITC-labeled albumin, and IgG on the morphology of crystals grown either slowly or fast. A rich variety of protein-specific and concentration-dependent morphologies was found and described by a morphological diagram. We give a phenomenological interpretation, which can explain the growth of complex patterns. Fluorescent images prove that a protein layer covers the surface of the KCl structures. We propose that this layer reduces the anisotropy of the effective surface tension during growth. The tip splitting fractal regime is attributed to the decrease of anisotropy. Further possible mechanisms, which can cause a morphological transition, are also discussed. We found elongated saw toothed crystals induced by proteins, especially IgG, and identified their structure. PMID- 12636534 TI - Molecular dynamics simulations of hydrated unsaturated lipid bilayers in the liquid-crystal phase and comparison to self-consistent field modeling. AB - Molecular dynamics simulations, using the collision dynamics method, were carried out for hydrated bilayers of 1-stearoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (18:0/18:1 omega 9cis PC, SOPC) and 1-stearoyl-2-docosahexaenoyl-sn-glycero-3 phosphatidylcholine (18:0/22:6 omega 3cis PC, SDPC). The simulation cells of the two bilayers consisted of 96 SOPC (or SDPC) molecules and 2304 water molecules: 48 lipid molecules per layer and 24 H2O molecules per lipid. The water was modeled by explicit TIP3P water molecules. The C-H bond-order-parameter -S(CH) profiles of the hydrocarbon tails, the bond orientation distribution functions and the root-mean-square values of the positional fluctuations of the lipid chain carbons were calculated. Simulation results are compared to the available experimental data and to other computer investigations of these lipid molecules. Several results of molecular-level self-consistent field calculations of these bilayers are also presented. Both theoretical methods reveal the same main characteristic features of the order-parameter profiles for the given bilayers. Some aspects of the physical properties of unsaturated lipids and their biological significance are discussed. PMID- 12636535 TI - Self-consistent-field modeling of hydrated unsaturated lipid bilayers in the liquid-crystal phase and comparison to molecular dynamics simulations. AB - A molecular-level self-consistent-field (SCF) theory is applied to model the lipid bilayer structures composed of 1-stearoyl-2-oleoyl-sn-glycero-3 phosphatidylcholine (18:0/18:1 omega 9cis PC) and 1-stearoyl-2-docosahexaenoyl-sn glycero-3-phosphatidylcholine (18:0/22:6 omega 3cis PC). As compared to earlier attempts to model (saturated) PC membranes several additional features are implemented: (i) A water model is used which correctly leads to low water concentration in the bilayers. (ii) Free volume is allowed for, which is important to obtain bilayers in the fluid state. (iii) A polarization term is included in the segment potentials; this new feature corrects for a minor thermodynamic inconsistency present in (all) earlier results for charged bilayers. (iv) The CH3 groups in the lipid molecules are assumed to have twice the volume of a CH2 group; this leads to stable noninterdigitated bilayers. (v) A cis double bond is simulated by forcing gauche conformations along the sn-2 acyl chain. Results of an all-atom molecular dynamics (MD) simulation, using the collision dynamics method, on the same system are presented. Both SCF and MD prove, in accordance with experimental facts, that acyl unsaturation effectively reduces the length of the chain which counteracts interdigitation. It is also found that the phosphatidylcholine head group is lying almost flat on the membrane surface and the water penetrates into the bilayer upto the glycerol backbone units. From the SCF results it further followed that the free volume is not exactly evenly distributed over the bilayer. There is a small increase in free volume in the center of the bilayer as well as in the glycerol backbone region. PMID- 12636536 TI - Theory of the effect of unsaturation on the main-chain transition. AB - We calculate the temperature of the main transition of lipids as a function of chain length for saturated lipids, and as a function of location of a cis double bond for lipids with one or two monounsaturated chains. We employ an extension of Marcelja's self-consistent field method which contains only two interaction parameters, and include many more chain orientations. Agreement with experiment on both the variation with chain length and with location of the double bond is satisfactory. PMID- 12636537 TI - Glass transition in an off-lattice protein model studied by molecular dynamics simulations. AB - In this paper we report the results of a numerical investigation of the glass transition phenomenon in a minimalist protein model. The inherent structure theory of Stillinger and Weber was applied to an off-lattice protein model with a native state beta-sheet motif. By using molecular dynamics simulations and the steepest descent method, sets of local potential energy minima were generated for the model over a range of temperatures. The mean potential energy of the inherent structures allowed to make rough estimates of the glass-transition temperature T(K). More accurately T(K) was computed by direct evaluations of the total and vibrational entropies. It is found that for the present model the thermodynamic ratio of the folding and glass-transition temperatures is 1.7 which is in good agreement with experimental observations. PMID- 12636538 TI - Quasistatic scale-free networks. AB - A network is formed using the N sites of a one-dimensional lattice in the shape of a ring as nodes and each node with the initial degree k(in)=2. N links are then introduced to this network, each link starts from a distinct node, the other end being connected to any other node with degree k randomly selected with an attachment probability proportional to k(alpha). Tuning the control parameter alpha, we observe a transition where the average degree of the largest node changes its variation from N0 to N at a specific transition point of alpha(c). The network is scale free, i.e., the nodal degree distribution has a power law decay for alpha> or = alpha(c). PMID- 12636539 TI - Evaluating surface tension using grand-canonical transition-matrix Monte Carlo simulation and finite-size scaling. AB - This Brief Report describes an approach for determining the surface tension of a model system that is applicable over the entire liquid-vapor coexistence region. At the heart of the method is a technique for determining coexistence properties that utilize transition probabilities of attempted Monte Carlo moves during a grand canonical simulation. Finite-size scaling techniques are implemented to determine the infinite system surface tension from a series of finite-size simulations. To demonstrate the method, the surface tension of the Lennard-Jones fluid is determined at temperatures ranging from the triple point to the critical point. PMID- 12636540 TI - Collective dynamics in liquid lithium, sodium, and aluminum. AB - Inelastic x-ray scattering data of liquid Li, Na, and Al for dynamical structure factors have been analyzed by proposing a semiempirical model. The model is based on the extension of the hydrodynamic model to the viscoelastic region so that it satisfies the first four nonvanishing sum rules. It has been found that the semiempirical model fits well with the x-ray scattering data for liquid metals investigated here. The physical meaning of the parameters is also discussed. PMID- 12636541 TI - Two-dimensional condensed phases from particles with tunable interactions. AB - We present a conceptually simple experimental model for condensed phases, consisting of an ensemble of identical magnetic dipoles on a vibrating bed. The model combines tunable and accurately known pair potentials, equilibration times of seconds, and lattice structure and dynamics visible to the naked eye. Fundamental ensemble properties--specifically phonon propagation, edge relaxation, and binary condensation--are directly observed and quantitatively linked to the underlying pair potential. PMID- 12636542 TI - Scaling of the nucleation density for pulsed layer deposition. AB - We discuss the logarithmic scaling of the nucleation density for pulsed laser deposition, discovered recently by Hinnemann et al. [Phys. Rev. Lett. 87, 135701 (2001)] in two dimensions. The logarithmic scaling is often observed in the upper critical dimension. We find that the nucleation density in one dimension also exhibits logarithmic scaling, implying that it is not a prerequisite for the upper critical dimension. The normalized island density also scales similarly both in one and two dimensions when plotted against the normalized coverage. PMID- 12636543 TI - Bending elasticity and bending fluctuations of lipid bilayer containing an additive. AB - The static and dynamic behavior of a bilayer containing an additive is examined theoretically. We have proved that the amplitudes of the thermal shape fluctuations of a quasi spherical lipid vesicle depend on the value of the bending elasticity of the vesicle's membrane at free exchange of molecules between its constituent monolayers. The dependence is calculated of this bending elasticity as a function of the concentration of the additive in the low concentration domain. In the same domain, the autocorrelation function of each vesicle fluctuation mode is found to be dependent on the two-dimensional diffusion coefficient of the bilayer additive. PMID- 12636544 TI - Resilience to damage of graphs with degree correlations. AB - The existence or nonexistence of a percolation threshold on power law correlated graphs is a fundamental question for which a general criterion is lacking. In this work we investigate the problems of site and bond percolation on graphs with degree correlations and their connection with spreading phenomena. We obtain some general expressions that allow the computation of the transition thresholds or their bounds. Using these results we study the effects of assortative and disassortative correlations on the resilience to damage of networks. PMID- 12636545 TI - Nonequilibrium thermodynamic description of the coupling between structural and entropic modes in supercooled liquids. AB - The density response of supercooled glycerol to an impulsive stimulated thermal grating (q=0.63 microm(-1)) has been studied in the temperature range (T=200-340 K) where the structure rearrangement (alpha relaxation) and the thermal diffusion occur on the same time scale. A strong interaction between the two modes occurs giving rise to a dip in the T dependence of the apparent thermal conductivity and a flattening of the apparent alpha-relaxation time upon cooling. A nonequilibrium thermodynamic model for the long time response has been developed. The model is capable to reproduce the experimental data and to explain the observed phenomenology. PMID- 12636546 TI - Universality and double critical end points. AB - A double critical end point in the two-dimensional spin-3/2 Blume-Capel model is studied via extensive Monte Carlo simulations. The resultant scaling character of the probability distribution of the mixing scaling operators allows us to locate the double critical end point precisely and also to convincingly show that it indeed belongs to the same universality class as the critical points. PMID- 12636547 TI - Chaos-assisted instanton tunneling in one-dimensional perturbed periodic potential. AB - For the system with a one-dimensional spatially periodic potential we demonstrate that small periodic in-time perturbation results in the appearance of chaotic instanton solutions. We estimate the parameter of local instability, the width of the stochastic layer, and the correlator for perturbed instanton solutions. The application of the instanton technique enables us to calculate the amplitude of the tunneling, the form of the spectrum, and the lower bound for the width of the ground quasienergy zone. PMID- 12636548 TI - Competition between two frequencies for phase synchronization of a chaotic laser. AB - Competition between two distinct driving frequencies to phase synchronize the intensity dynamics of a chaotic laser has been observed. The phase of the chaotic intensity signal is constructed using the complex analytic signal. Competing frequencies alternately show phase locking and phase slipping. Competition has been quantified by calculating the portion of time the laser phase locks to each of the driving frequencies and their average. PMID- 12636549 TI - Anomalous heat conduction in a one-dimensional ideal gas. AB - We provide firm convincing evidence that the energy transport in a one dimensional gas of elastically colliding free particles of unequal masses is anomalous, i.e., the Fourier law does not hold. Our conclusions are confirmed by a theoretical and numerical analysis based on a Green-Kubo-type approach specialized to momentum-conserving lattices. PMID- 12636550 TI - Enhancement of noise-induced escape through the existence of a chaotic saddle. AB - We study the noise-induced escape process in a prototype dissipative nonequilibrium system, the Ikeda map. In the presence of a chaotic saddle embedded in the basin of attraction of the metastable state, we find the novel phenomenon of a strong enhancement of noise-induced escape. This result is established by employing the theory of quasipotentials. Our finding is of general validity and should be experimentally observable. PMID- 12636551 TI - Noise-enhanced synchronization of homoclinic chaos in a CO2 laser. AB - Many chaotic oscillators have rather coherent phase dynamics but strong fluctuation in the amplitudes. Conversely, homoclinic chaos is characterized by quite regular spikes but strong fluctuation in their time intervals. We study the effects of noise on the synchronization of homoclinic chaos to a weak periodic signal and demonstrate numerically and experimentally in a CO2 laser system that noise enhances synchronization of homoclinic chaos. The system exhibits both conventional resonance versus driving frequency and stochastic resonance with respect to noise intensity. PMID- 12636552 TI - Coupled replicator equations for the dynamics of learning in multiagent systems. AB - Starting with a group of reinforcement-learning agents we derive coupled replicator equations that describe the dynamics of collective learning in multiagent systems. We show that, although agents model their environment in a self-interested way without sharing knowledge, a game dynamics emerges naturally through environment-mediated interactions. An application to rock-scissors-paper game interactions shows that the collective learning dynamics exhibits a diversity of competitive and cooperative behaviors. These include quasiperiodicity, stable limit cycles, intermittency, and deterministic chaos behaviors that should be expected in heterogeneous multiagent systems described by the general replicator equations we derive. PMID- 12636553 TI - Lasing on scar modes in fully chaotic microcavities. AB - Scar wave functions in a fully chaotic cavity are obtained numerically by an extended Fox-Li method. Lasing on the scar modes are observed in a semiconductor microcavity with a selective excitation of different scars controlled by corresponding shape of electrodes for current injection. PMID- 12636554 TI - Third-order harmonic generation by self-guided femtosecond pulses in air. AB - Strong third-order harmonic (TH) emission is observed with a conversion efficiency higher than 10(-3) from a plasma channel formed by self-guided femtosecond laser pulses propagating in air. The main characteristics of TH emission in various conditions and the phase-matching condition between the fundamental and the TH wave are investigated. An optimized condition is found, under which the TH conversion efficiency is maximized. Our experimental results show that radiation of the emission in ultraviolet wavelength range makes a major attribution to TH emission, whereas the effects of self-phase modulation are not important when intense laser pulses interact with gaseous media. PMID- 12636555 TI - Search for the prewave zone effect in transition radiation. AB - Results of the first experimental search for the effect of the prewave zone in near-infrared transition radiation are presented. A substantial difference in the spatial distribution of transition relation for two different wavelengths (450 nm and 1600 nm) was observed. Experimental data are in a good, though not complete, agreement with the theory. PMID- 12636556 TI - Saddle-node bifurcation: appearance mechanism of pulses in the subcritical complex Ginzburg-Landau equation. AB - We study stationary, localized solutions in the complex subcritical Ginzburg Landau equation in the region where there exists coexistence of homogeneous attractors. Using a matching approach, we report on the fact that the appearance of pulses are related to a saddle-node bifurcation. Numerical simulations are in good agreement with our theoretical predictions. PMID- 12636557 TI - Cluster Monte Carlo algorithm for the quantum rotor model. AB - We propose a highly efficient "worm"-like cluster Monte Carlo algorithm for the quantum rotor model in the link-current representation. We explicitly prove detailed balance for the algorithm even in the presence of disorder. For the pure quantum rotor model with mu=0, the algorithm yields high- precision estimates for the critical point K(c)=0.333 05(5) and the correlation length exponent nu=0.670(3). For the disordered case, mu=1 / 2+/-1 / 2, we find nu=1.15(10). PMID- 12636558 TI - Stochastic processes and conformal invariance. AB - We discuss a one-dimensional model of a fluctuating interface with a dynamic exponent z=1. The events that occur are adsorption, which is local, and desorption which is nonlocal and may take place over regions of the order of the system size. In the thermodynamic limit, the time dependence of the system is given by characters of the c=0 logarithmic conformal field theory of percolation. This implies in a rigorous way, a connection between logarithmic conformal field theory and stochastic processes. The finite-size scaling behavior of the average height, interface width and other observables are obtained. The avalanches produced during desorption are analyzed and we show that the probability distribution of the avalanche sizes obeys finite-size scaling with new critical exponents. PMID- 12636559 TI - Computing a non-Maxwellian velocity distribution from first principles. AB - We investigate a family of single-particle anomalous velocity distribution by solving a particular class of stochastic Liouville equations. The stationary state is obtained analytically and the Maxwell-Boltzmann distribution is reobtained in a particular limit. We discuss the comparison with other different methods to obtain the stationary state. Extensions when the models cannot be solved in an exact way are also pointed out in connection with the one-ficton approximation. PMID- 12636560 TI - Dynamic rigidity transition. AB - An inflated closed loop (or membrane) is used to demonstrate a dynamic rigidity transition that occurs when impact energy is added to the loop in static equilibrium at zero temperature. The only relevant parameter in this transition is the ratio of the energy needed to collapse the loop and the impact energy. When this ratio is below a threshold value close to unity, the loop collapses into a high-entropy floppy state, and it does not return to the rigid state unless the impact energy can escape. The internal oscillations are in the floppy state dominated by 1/f(2) noise. When the ratio is above the threshold, the loop does not collapse, and the internal oscillations resulting from the impact are dominated by the eigenfrequencies of the stretched membrane. In this state, the loop can bounce for a long time. It is still an open question whether bouncing will eventually vanish or whether a stationary bouncing state will be reached. The dynamic transition between the floppy and the rigid state is discontinuous. PMID- 12636561 TI - Family of additive entropy functions out of thermodynamic limit. AB - We derive a one-parametric family of entropy functions that respect the additivity condition, and which describe effects of finiteness of statistical systems, in particular, distribution functions with long tails. This one parametric family is different from the Tsallis entropies, and is a convex combination of the Boltzmann-Gibbs-Shannon entropy and the entropy function proposed by Burg. An example of how longer tails are described within the present approach is worked out for the canonical ensemble. We also discuss a possible origin of a hidden statistical dependence, and give explicit recipes on how to construct corresponding generalizations of the master equation. PMID- 12636562 TI - Approximate calculation of the ground-state energy for Potts spin-glass models. AB - We consider the q-state Potts spin-glass model, with quenched couplings taking two different values only. As an approximation for this model a proper generalization of the random energy model is derived. Formulas of the resulting diluted generalized random energy model (DGREM) are applied to calculate the ground-state energy for the two-dimensional Potts spin-glass model. The semianalytical results are compared with numerical determinations of the ground state energy, using multicanonical, random cost, and simulated annealing techniques. PMID- 12636563 TI - Itineration of the Internet over nonequilibrium stationary states in Tsallis statistics. AB - The cumulative probability distribution of sparseness time interval in the Internet is studied by the method of data analysis. Round-trip time between a local host and a destination host through ten odd routers is measured using the ping command, i.e., doing an echo experiment. The data are found to be well described by q-exponential distributions, which maximize the Tsallis entropy indexed by q less or larger than unity, showing a scale-invariant feature of the system. The network is observed to itinerate over a series of the nonequilibrium stationary states characterized by Tsallis statistics. PMID- 12636565 TI - Variational methods for the solution of the Ornstein-Zernicke equation in inhomogeneous systems. AB - We show that the Ornstein-Zernicke equation and other equations of similar form obey a variational principle that can be used to derive approximate solutions. This method requires the use of an initial trial solution where the variational solution possesses a stationary "point" with respect to the trial solution when the latter is equal to the exact solution. We show that with even a very simple form of the trial solution the results are quite reasonable. Furthermore, we have demonstrated that by combining the variational method with an iterative expansion of the Ornstein-Zernicke equation it is possible to develop a self-consistent method of writing the direct correlation function. PMID- 12636564 TI - Phase diagram of a probabilistic cellular automaton with three-site interactions. AB - We study a (1+1)-dimensional probabilistic cellular automaton that is closely related to the Domany-Kinzel stochastic-cellular automaton (DKCA), but in which the update of a given site depends on the state of three sites at the previous time step. Thus, compared with the DKCA, there is an additional parameter p(3) representing the probability for a site to be active at time t, given that it and its nearest neighbors were active at time t-1. We study phase transitions and critical behavior for the activity and for damage spreading, using one- and two site mean-field approximations, and simulations, for p(3)=0 and p(3)=1. We find evidence for a line of tricritical points in the (p(1),p(2),p(3)) parameter space, obtained using a mean-field approximation at pair level. To construct the phase diagram in simulations we employ the growth-exponent method in an interface representation. For p(3)=0, the phase diagram is similar to the DKCA, but the damage-spreading transition exhibits a reentrant phase. For p(3)=1, the growth exponent method reproduces the two absorbing states, first- and second-order phase transitions, bicritical point, and damage-spreading transition recently identified by Bagnoli et al. [Phys. Rev. E 63, 046116 (2001)]. PMID- 12636566 TI - Temporal oscillations and phase transitions in the evolutionary minority game. AB - The study of societies of adaptive agents seeking minority status is an active area of research. Recently, it has been demonstrated that such systems display an intriguing phase transition: agents tend to self-segregate or to cluster according to the value of the prize-to-fine ratio R. We show that such systems do not establish a true stationary distribution. The winning probabilities of the agents display temporal oscillations. The amplitude and frequency of the oscillations depend on the value of R. The temporal oscillations that characterize the system explain the transition in the global behavior from self segregation to clustering in the R<1 case. PMID- 12636567 TI - Logarithmic roughening in a growth process with edge evaporation. AB - Roughening transitions are often characterized by unusual scaling properties. As an example we investigate the roughening transition in a solid-on-solid growth process with edge evaporation [U. Alon, M. Evans, H. Hinrichsen, and D. Mukamel, Phys. Rev. Lett. 76, 2746 (1996)], where the interface is known to roughen logarithmically with time. Performing high-precision simulations we find appropriate scaling forms for various quantities. Moreover we present a simple approximation explaining why the interface roughens logarithmically. PMID- 12636568 TI - Critical behavior of the one-dimensional diffusive pair contact process. AB - The phase transition of the one-dimensional diffusive pair contact process is investigated by N cluster mean-field approximations and high precision simulations. The N=3,4 cluster approximations exhibit smooth transition line to absorbing state by varying the diffusion rate D with beta(2)=2 mean-field order parameter exponent of the pair density. This contradicts with former N=2 results, where two different mean-field behavior was found along the transition line. Extensive dynamical simulations on L=10(5) lattices give estimates for the order parameter exponents of the particles for 0.050 reaction taking place on a one-dimensional chain of length N (N--> infinity ) in which some segments (placed at random, with mean concentration p) possess special, catalytic properties. Annihilation reaction takes place as soon as any two A particles land onto two vacant sites at the extremities of the catalytic segment, or when any A particle lands onto a vacant site on a catalytic segment while the site at the other extremity of this segment is already occupied by another A particle. Noncatalytic segments are inert with respect to reaction and here two adsorbed A particles harmlessly coexist. For both "annealed" and "quenched" disorder in placement of the catalytic segments, we calculate exactly the disorder-averaged pressure per site. Explicit asymptotic formulas for the particle mean density and the compressibility are also presented. PMID- 12636573 TI - Anomalous diffusion and Hall effect on comb lattices. AB - In this paper we study the effects of a magnetic field on the discrete time random walk of a classical charged particle moving on a comb lattice. We develop an analytical technique to study the Lorentz force effects on the asymptotic diffusion laws. This approach also allows the description of the combined action of an electric and a magnetic field (Hall effect). The generalization to other comblike branched structures is discussed. PMID- 12636575 TI - Bistable gradient networks. I. Attractors and pattern retrieval at low loading in the thermodynamic limit. AB - We examine the large-network, low-loading behavior of an attractor neural network, the so-called bistable gradient network (BGN), and compare it with that of the Hopfield network (HN). We use analytical and numerical methods to characterize the attractor states of the network and their basins of attraction. The energy landscape of BGN is more complex than that of the HN and depends on the strength of the coupling among units. At weak coupling, the BGN acts as a highly selective associative memory; the input must be close to the one of the stored patterns in order to be recognized. A category of spurious attractors occurs which is not present in the HN. Stronger coupling results in a transition to a more Hopfield-like regime with large basins of attraction. The basins of attraction for spurious attractors are noticeably suppressed compared to the Hopfield case, even though the Hebbian synaptic structure is the same and there is no stochastic noise. PMID- 12636574 TI - Packing-limited growth of irregular objects. AB - We study growth limited by packing for irregular objects in two dimensions. We generate packings by seeding objects randomly in time and space and allowing each object to grow until it collides with another object. The objects we consider allow us to investigate the separate effects of anisotropy and nonunit aspect ratio. By means of a connection to the decay of pore-space volume, we measure power law exponents for the object size distribution. We carry out a scaling analysis, showing that it provides an upper bound for the size distribution exponent. We find that while the details of the growth mechanism are irrelevant, the exponent is strongly shape dependent. Potential applications lie in ecological and biological environments where sessile organisms compete for limited space as they grow. PMID- 12636576 TI - Bistable gradient networks. II. Storage capacity and behavior near saturation. AB - We examine numerically the storage capacity and the behavior near saturation of an attractor neural network consisting of bistable elements with an adjustable coupling strength, the bistable gradient network. For strong coupling, we find evidence of a first-order "memory blackout" phase transition, as in the Hopfield network. For weak coupling, on the other hand, there is no evidence of such a transition and memorized patterns can be stable even at high levels of loading. The enhanced storage capacity comes, however, at the cost of imperfect retrieval of the patterns from corrupted versions. PMID- 12636577 TI - Signal detection via residence-time asymmetry in noisy bistable devices. AB - We introduce a dynamical readout description for a wide class of nonlinear dynamic sensors operating in a noisy environment. The presence of weak unknown signals is assessed via the monitoring of the residence time in the metastable attractors of the system, in the presence of a known, usually time-periodic, bias signal. This operational scenario can mitigate the effects of sensor noise, providing a greatly simplified readout scheme, as well as significantly reduced processing procedures. Such devices can also show a wide variety of interesting dynamical features. This scheme for quantifying the response of a nonlinear dynamic device has been implemented in experiments involving a simple laboratory version of a fluxgate magnetometer. We present the results of the experiments and demonstrate that they match the theoretical predictions reasonably well. PMID- 12636578 TI - Functional renormalization group for anisotropic depinning and relation to branching processes. AB - Using the functional renormalization group, we study the depinning of elastic objects in presence of anisotropy. We explicitly demonstrate how the Kardar Parisi-Zhang (KPZ) term is always generated, even in the limit of vanishing velocity, except where excluded by symmetry. This mechanism has two steps. First a nonanalytic disorder-distribution is generated under renormalization beyond the Larkin length. This nonanalyticity then generates the KPZ term. We compute the beta function to one loop taking properly into account the nonanalyticity. This gives rise to additional terms, missed in earlier studies. A crucial question is whether the nonrenormalization of the KPZ coupling found at 1-loop order extends beyond the leading one. Using a Cole-Hopf-transformed theory we argue that it is indeed uncorrected to all orders. The resulting flow equations describe a variety of physical situations: We study manifolds in periodic disorder, relevant for charge density waves, as well as in nonperiodic disorder. Further the elasticity of the manifold can either be short range (SR) or long range (LR). A careful analysis of the flow yields several nontrivial fixed points. All these fixed points are transient since they possess one unstable direction towards a runaway flow, which leaves open the question of the upper critical dimension. The runaway flow is dominated by a Landau-ghost mode. For LR elasticity, relevant for contact line depinning, we show that there are two phases depending on the strength of the KPZ coupling. For SR elasticity, using the Cole-Hopf transformed theory we identify a nontrivial 3-dimensional subspace which is invariant to all orders and contains all above fixed points as well as the Landau mode. It belongs to a class of theories which describe branching and reaction-diffusion processes, of which some have been mapped onto directed percolation. PMID- 12636579 TI - Chaos based on Riemannian geometric approach to Abelian-Higgs dynamical system. AB - Based on the Riemannian geometric approach, we study chaos of the Abelian-Higgs dynamical system derived from a classical field equation consisting of a spatially homogeneous Abelian gauge field and Higgs field. Using the global indicator of chaos formulated by the sectional curvature of the ambient manifold, we show that this approach brings the same qualitative and quantitative information about order and chaos as has been provided by the Lyapunov exponents in the conventional and phenomenological approach. We confirm that the mechanism of chaos is a parametric instability of the system. By analyzing a close relation between the sectional curvature and the Gaussian curvature, we point out that the Toda-Brumer criterion becomes a sufficient condition to the criterion based on this geometric approach as to the stability condition. PMID- 12636580 TI - Properties and features of asymmetric partial devil's staircases deduced from piecewise linear maps. AB - A piecewise linear map with one discontinuity is used to link together iterated map properties with the shape of the ensuing staircases. In the main part of the paper, a three-segment map is treated, with a horizontal middle segment next to the discontinuity and the development of partial and asymmetric staircases is demonstrated. In particular, a possible hierarchy of partiality, connected with the ratio of the length of the horizontal segment to the discontinuity jump, is obtained. The map is used for constructing staircases that imitate various experimental and numerical staircases that appear in the literature for excitable systems. PMID- 12636581 TI - Application of computational mechanics to the analysis of natural data: an example in geomagnetism. AB - We discuss how the ideal formalism of computational mechanics can be adapted to apply to a noninfinite series of corrupted and correlated data, that is typical of most observed natural time series. Specifically, a simple filter that removes the corruption that creates rare unphysical causal states is demonstrated, and the concept of effective soficity is introduced. We believe that computational mechanics cannot be applied to a noisy and finite data series without invoking an argument based upon effective soficity. A related distinction between noise and unresolved structure is also defined: Noise can only be eliminated by increasing the length of the time series, whereas the resolution of previously unresolved structure only requires the finite memory of the analysis to be increased. The benefits of these concepts are demonstrated in a simulated times series by (a) the effective elimination of white noise corruption from a periodic signal using the expletive filter and (b) the appearance of an effectively sofic region in the statistical complexity of a biased Poisson switch time series that is insensitive to changes in the word length (memory) used in the analysis. The new algorithm is then applied to an analysis of a real geomagnetic time series measured at Halley, Antarctica. Two principal components in the structure are detected that are interpreted as the diurnal variation due to the rotation of the Earth-based station under an electrical current pattern that is fixed with respect to the Sun Earth axis and the random occurrence of a signature likely to be that of the magnetic substorm. In conclusion, some useful terminology for the discussion of model construction in general is introduced. PMID- 12636582 TI - Bifurcation scenarios for bubbling transition. AB - Dynamical systems with chaos on an invariant submanifold can exhibit a type of behavior called bubbling, whereby a small random or fixed perturbation to the system induces intermittent bursting. The bifurcation to bubbling occurs when a periodic orbit embedded in the chaotic attractor in the invariant manifold becomes unstable to perturbations transverse to the invariant manifold. Generically the periodic orbit can become transversely unstable through a pitchfork, transcritical, period-doubling, or Hopf bifurcation. In this paper a unified treatment of the four types of bubbling bifurcation is presented. Conditions are obtained determining whether the transition to bubbling is soft or hard; that is, whether the maximum burst amplitude varies continuously or discontinuously with variation of the parameter through its critical value. For soft bubbling transitions, the scaling of the maximum burst amplitude with the parameter is derived. For both hard and soft transitions the scaling of the average interburst time with the bifurcation parameter is deduced. Both random (noise) and fixed (mismatch) perturbations are considered. Results of numerical experiments testing our theoretical predictions are presented. PMID- 12636584 TI - Semiclassical theory for transmission through open billiards: convergence towards quantum transport. AB - We present a semiclassical theory for transmission through open quantum billiards which converges towards quantum transport. The transmission amplitude can be expressed as a sum over all classical paths and pseudopaths which consist of classical path segments joined by "kinks," i.e., diffractive scattering at lead mouths. For a rectangular billiard we show numerically that the sum over all such paths with a given number of kinks K converges to the quantum transmission amplitude as K--> infinity. Unitarity of the semiclassical theory is restored as K approaches infinity. Moreover, we find excellent agreement with the quantum path-length power spectrum up to very long path length. PMID- 12636585 TI - Patterns of electroconvection in the nematic liquid crystal N4. AB - Electroconvection using the liquid crystal N4 is studied as a function of two control parameters: the applied frequency and the applied voltage. As a function of voltage, there is a rich series of bifurcations that takes the system from stationary rolls to chaos. As a function of the frequency, the initial pattern changes from stationary oblique rolls at low frequencies to stationary normal rolls at higher frequencies. There is also a change in the secondary bifurcations. In particular, we observe that the bimodal-varicose instability is replaced by the skewed-varicose instability as the applied frequency is increased. Comparisons with theoretical predictions are made. PMID- 12636583 TI - Short-time decay of the Loschmidt echo. AB - The Loschmidt echo measures the sensitivity to perturbations of quantum evolutions. We study its short-time decay in classically chaotic systems. Using perturbation theory and throwing out all correlation imposed by the initial state and the perturbation, we show that the characteristic time of this regime is well described by the inverse of the width of the local density of states. This result is illustrated and discussed in a numerical study in a two-dimensional chaotic billiard system perturbed by various contour deformations and using different types of initial conditions. Moreover, the influence to the short-time decay of sub-Planck structures developed by time evolution is also investigated. PMID- 12636586 TI - Relevance of symmetry for the synchronization of chaotic optical systems and the related Lang-Kobayashi model limitations. AB - Synchronization of chaotic semiconductor lasers has now been demonstrated experimentally in a variety of coupling schemes. Coupling methods include configurations where the transmitter laser system is itself chaotic and drives a receiver system, both lasers are individually chaotic, and both lasers induce the chaos through mutual coupling. The dynamics for each of these scenarios is in many cases adequately captured by the Lang-Kobayashi rate equation model. Such a simplified model, however, ignores fundamental aspects of the laser dynamics, such as the frequency and carrier density material susceptibility dependence, spatial hole burning effects, proper boundary conditions, and the fact that lasers may exhibit pronounced multilongitudinal dynamic behavior with and without the presence of a weak external feedback or injection. The model also cannot distinguish between many of the possible coupling geometries realizable in experiments. Using an interactive simulator based on the rigorous microscopic description of the light-matter interaction, we explore the unidirectionally coupled configuration, the relevance of symmetry for the synchronization achieved between two identical lasers, and the differences that arise when the traditional analysis through the Lang-Kobayashi model is compared to the full nonlinear partial differential equation model results. PMID- 12636587 TI - Use of chaotic excitation and attractor property analysis in structural health monitoring. AB - This work explores the utility of attractor-based approaches in the field of vibration-based structural health monitoring. The technique utilizes the unique properties of chaotic signals by driving the structure directly with the output of a chaotic oscillator. Using the Kaplan-Yorke conjecture, the Lyapunov exponents of the driving signal may be tuned to the dominant eigenvalues of the structure, thus controlling the dimension of the structural response. Data are collected at various stages of structural degradation and a simple nonlinear model, constructed from the undamaged data, is used to make predictions for the damaged response data. Prediction error is then introduced as a "feature" for classifying the magnitude of the damage. Results are presented for an experimental cantilevered beam instrumented with fiber-optic strain sensors. PMID- 12636588 TI - Cooperative dynamics in a class of coupled two-dimensional oscillators. AB - We study a system of globally coupled two-dimensional nonlinear oscillators [using the two-junction superconducting quantum interference device (SQUID) as a prototype for a single element] each of which can undergo a saddle-node bifurcation characterized by the disappearance of the stable minima in its potential energy function. This transition from fixed point solutions to spontaneous oscillations is controlled by external bias parameters, including the coupling coefficient. For the deterministic case, an extension of a center manifold reduction, carried out earlier for the single oscillator, yields an oscillation frequency that depends on the coupling; the frequency decreases with coupling strength and/or the number of oscillators. In the presence of noise, a mean-field description leads to a nonlinear Fokker-Planck equation for the system which is investigated for experimentally realistic noise levels. Furthermore, we apply a weak external time-sinusoidal probe signal to each oscillator and use the resulting (classical) resonance to determine the underlying frequency of the noisy system. This leads to an explanation of earlier experimental results as well as the possibility of designing a more sensitive SQUID-based detection system. PMID- 12636589 TI - Uniform semiclassical wave function for coherent two-dimensional electron flow. AB - We find a uniform semiclassical (SC) wave function describing coherent branched flow through a two-dimensional electron gas (2DEG), a phenomenon recently discovered by direct imaging of the current using scanned probed microscopy [M.A. Topinka, B.J. LeRoy, S.E.J. Shaw, E.J. Heller, R.M. Westervelt, K.D. Maranowski, and A.C. Gossard, Science 289, 2323 (2000)]. The formation of branches has been explained by classical arguments [M.A. Topinka, B.J. LeRoy, R.M. Westervelt, S.E.J. Shaw, R. Fleischmann, E.J. Heller, K.D. Maranowski, and A.C. Gossard, Nature (London) 410, 183 (2001)], but the SC simulations necessary to account for the coherence are made difficult by the proliferation of catastrophes in the phase space. In this paper, expansion in terms of "replacement manifolds" is used to find a uniform SC wave function for a cusp singularity. The method is then generalized and applied to calculate uniform wave functions for a quantum-map model of coherent flow through a 2DEG. Finally, the quantum-map approximation is dropped and the method is shown to work for a continuous-time model as well. PMID- 12636590 TI - Synchronization and intermittency in three-coupled chaotic oscillators. AB - Synchronization of three-coupled chaotic oscillators was studied with the use of a coupled map system derived for interacting kicked relaxators. Partial synchronization (PS), in which two of the three were synchronized, was observed in addition to complete synchronization. An intermittency associated with the breakdown of the PS, seemingly different from the conventional on-off intermittency, was found. We elucidated the statistics, observing the burst-size distribution, the laminar duration distribution, etc. It was found that the breakdown of the PS generated an anomalous diffusion different from that associated with on-off intermittency. PMID- 12636592 TI - Analytical approach to the drift of the tips of spiral waves in the complex Ginzburg-Landau equation. AB - In this paper, we investigate the motion of spiral waves in the complex Ginzburg Landau equation (CGLE) analytically and numerically. We find that the tip of the spiral wave drifts primarily in the direction of the electric field and there is a smaller component of the drift that is perpendicular to the field when a uniform field is applied to the system. The velocity of the tip is uniform and its component along the electric field is equal to the strength of the field. When the CGLE system is driven by white noise, a diffusion law for the vortex core of the spiral wave is derived at long time explicitly. The diffusion constant is found to be D=T/C(2), in which T is the noise strength and C is the core asymptotic factor of the spiral wave. When the external force is a simple oscillation we find that the tip of the spiral wave drifts if the frequency of the external force is the same as that of the system. Our analytical results are verified using numerical simulations. PMID- 12636591 TI - Effect of initial conditions on the speed of reaction-diffusion fronts. AB - The effect of initial conditions on the speed of propagating fronts in reaction diffusion equations is examined in the framework of the Hamilton-Jacobi theory. We study the transition between quenched and nonquenched fronts both analytically and numerically for parabolic and hyperbolic reaction diffusion. Nonhomogeneous media are also analyzed and the effect of algebraic initial conditions is also discussed. PMID- 12636593 TI - Complex phase dynamics in coupled bursters. AB - The phenomenon of phase multistability in the synchronization of two coupled oscillatory systems typically arises when the systems individually display complex wave forms associated, for instance, with the presence of subharmonic components. Alternatively, phase multistability can be caused by variations of the phase velocity along the orbit of the individual oscillator. Focusing on the mechanisms underlying the appearance of phase multistability, the paper examines a variety of phase-locked patterns in the bursting behavior of a model of coupled pancreatic cells. In particular, we show how the number of spikes per train and the proximity of a neighboring equilibrium point can influence the formation of coexisting regimes. PMID- 12636594 TI - Quantum chaotic trajectories in integrable right triangular billiards. AB - Right triangular billiards are very simple systems that are completely integrable in classical mechanics for acute angle pairs (45 degrees,45 degrees ) and (30 degrees,60 degrees ). In quantum mechanics, the energy level spacing distribution of these billiards are neither Poisson-like nor Wigner-like. We use Bohm's formalism to calculate the trajectories, by numerical methods, for a particle inside these billiards. We use a linear combination of the first three energy states as the initial wave function. We show that a particle can have quasiperiodic or chaotic behavior, depending on its initial position in the billiards. PMID- 12636596 TI - Instability of convection in a fluid layer rotating about an oblique axis. AB - We analyze thermal convection in a fluid layer confined between isothermal horizontal boundaries at which the tangential component of the fluid stress vanishes. The layer rotates about an oblique, nearly vertical axis. Using a model set of equations for w, the horizontal planform of the vertical velocity component, and psi, a stream function related to a large-scale vertical vorticity field, we describe the instabilities of convection rolls. We show how the usual Kuppers-Lortz instability, which leads to a continual precession of the roll pattern, can be suppressed by the oblique rotation vector. Of particular interest is the small-angle instability of rolls, to perturbations in the form of rolls that are almost aligned with the primary rolls; at finite Prandtl number, this instability is not prevented by the horizontal component of the rotation vector, unless this component is sufficiently strong, in which case stability is confined to small-amplitude rolls near the marginal stability boundary. A one-dimensional instability leading to amplitude-modulated rolls is unaffected by the oblique rotation. Numerical simulations of the model equations are presented, which illustrate the instabilities analyzed. PMID- 12636595 TI - Characterization of the noise effect on weak synchronization. AB - We investigate the noise effect on weak synchronization in two coupled identical one-dimensional (1D) maps. Due to the existence of positive local transverse Lyapunov exponents, the weakly stable synchronous chaotic attractor (SCA) becomes sensitive with respect to the variation of noise intensity. To quantitatively characterize such noise sensitivity, we introduce a quantifier, called the noise sensitivity exponent (NSE). For the case of bounded noise, the values of the NSE are found to be the same as those of the exponent characterizing a parameter sensitivity of the weakly stable SCA in presence of a parameter mismatch between the two 1D maps. Furthermore, it is found that the scaling exponent for the average time spent near the diagonal for both the bubbling and riddling cases occurring in the regime of weak synchronization is given by the reciprocal of the NSE, as in the parameter-mismatching case. Consequently, both the noise and parameter mismatch have the same effect on the scaling behavior of the average characteristic time. PMID- 12636597 TI - Coarsening dynamics of dewetting films. AB - Lubrication theory for unstable thin liquid films on solid substrates is used to model the coarsening dynamics in the long-time behavior of dewetting films. The dominant physical effects that drive the fluid dynamics in dewetting films are surface tension and intermolecular interactions with the solid substrate. Instabilities in these films lead to rupture and other morphological changes that promote nonuniformity in the films. Following the initial instabilities, the films break up into near-equilibrium droplets connected by an ultrathin film. For longer times, the fluid will undergo a coarsening process in which droplets both move and exchange mass on slow time scales. The dynamics of this coarsening process will be obtained through the asymptotic reduction of the long-wave PDE governing the thin film to a set of ODEs for the evolution of the droplets. From this, a scaling law that governs the coarsening rate is derived. PMID- 12636598 TI - Experimental study on the dynamics of gas-fluidized beds. AB - Gas-fluidized fine powders display three regimes of fluidization: solidlike, fluidlike, and bubbling. We investigate, from both macroscopic and local measurements, the transition between these regimes. We show that the transition between the solidlike and the fluidlike regimes takes place along an interval of gas velocities in which transient active regions alternate with transient solid networks. Although in the apparently homogeneous fluidlike regime large amplitude bubbles are not perceived and the bed expands continuously with increasing gas flow, optical probe local measurements show the existence of mesoscale pseudoturbulent structures and short-lived voids, reminiscent of liquid-fluidized beds behavior, and whose characteristic temporal frequency increases with gas velocity. These mesostructures might be responsible for the fast diffusion measured in gas-fluidized beds. PMID- 12636599 TI - Active and passive fields in turbulent transport: the role of statistically preserved structures. AB - We have recently proposed that the statistics of active fields (which affect the velocity field itself) in well-developed turbulence are also dominated by the statistically preserved structures of auxiliary passive fields which are advected by the same velocity field. The statistically preserved structures are eigenmodes of eigenvalue 1 of an appropriate propagator of the decaying (unforced) passive field, or equivalently, the zero modes of a related operator. In this paper we investigate further this surprising finding via two examples of shell models, one akin to turbulent convection in which the temperature is the active scalar, and the other akin to magnetohydrodynamics in which the magnetic field is the active vector. In the first example, all the even correlation functions of the active and passive fields exhibit identical scaling behavior. The second example appears at first sight to be a counterexample: the statistical objects of the active and passive fields have entirely different scaling exponents. We demonstrate, nevertheless, that the statistically preserved structures of the passive vector dominate again the statistics of the active field, except that due to a dynamical conservation law the amplitude of the leading zero mode cancels exactly. The active vector is then dominated by the subleading zero mode of the passive vector. Our work thus suggests that the statistical properties of active fields in turbulence can be understood with the same generality as those of passive fields. PMID- 12636600 TI - Vortex tubes in velocity fields of laboratory isotropic turbulence: dependence on the Reynolds number. AB - The streamwise and transverse velocities are measured simultaneously in isotropic grid turbulence at relatively high Reynolds numbers Re(lambda) approximately 110 330. Using a conditional averaging technique, we extract typical intermittency patterns that are consistent with velocity profiles of a model for a vortex tube, i.e., Burgers vortex. The radii of the vortex tubes are several of the Kolmogorov length, regardless of the Reynolds number. Using the distribution of an interval between successive enhancements of a small-scale velocity increment, we study the spatial distribution of vortex tubes. The vortex tubes tend to cluster together. This tendency is increasingly significant with the Reynolds number. Using statistics of velocity increments, we also study the energetical importance of vortex tubes as a function of the scale. The vortex tubes are important over the background flow at small scales especially below the Taylor microscale. At a fixed scale, the importance is increasingly significant with the Reynolds number. PMID- 12636601 TI - Bidirectional solitons on water. AB - A theory of bidirectional solitons on water is developed by using an integrable Boussinesq surface-variable equation. We present an explicit transformation between the system and a member of the Ablowitz-Kaup-Newell-Segur system, and derive an exact multisoliton solution by using a Darboux transformation. The phase shifts and the maximum wave heights during the interaction are studied for two-soliton overtaking and head-on collisions. They agree with the Korteweg-de Vries solution for overtaking collision and the perturbation solution for head-on collision. PMID- 12636602 TI - Thin film of non-Newtonian fluid on an incline. AB - The slow flow of thin liquid films on solid surfaces is an important phenomenon in nature and in industrial processes, and an intensive effort has been made to investigate it. It is well known that the contact line of currents on an inclined surface may become unstable and then a pattern of "fingers" develops that affects the quality of the coatings. This instability has been intensively studied due to its relevance for the technology of various industrial processes. So far the theoretical and numerical research has been focused on Newtonian fluids, notwithstanding that often in the real situations as well as in the experiments, the rheology of the involved liquid is non-Newtonian. Using the lubrication approximation, we derive the governing equations for a current of a power law non Newtonian fluid on an inclined plane under the action of gravity and the viscous stresses. We show that surface tension effects can be included in the theory by a slight modification of the governing equations, that can then be used as a starting point to investigate the influence of rheology on the fingering instability and other phenomena of interest. We consider the one-dimensional case and we present three families of traveling wave solutions: two running downwards and the other upwards. PMID- 12636603 TI - Influence of attractive van der Waals interactions on the optimal excitations in thermocapillary-driven spreading. AB - Recent investigations of microfluidic flows have focused on manipulating the motion of very thin liquid films by modulating the surface tension through an applied streamwise temperature gradient. The extent to which the choice of contact line model affects the flow and stability of such thermocapillary-driven films is not completely understood. Regardless of the contact line model used, the linearized disturbance operator corresponding to the evolution of the film height is non-normal, and a generalized non-modal stability analysis is required. Surprisingly, early predictions of frontal instability that stemmed from conventional modal analysis of thermocapillary flow on a flat, infinite precursor film showed excellent agreement with experiment. Within the more rigorous framework provided by a generalized stability analysis, this work investigates the transient dynamics and amplification of optimal disturbances subject to a finite precursor film generated by attractive van der Waals forces. Convergence of the disturbance growth rates and perturbed shapes to the asymptotic solutions obtained by conventional linear stability analysis occurs early in the spreading process. In addition, the level of transient disturbance amplification is minimal. The equations governing thermocapillary-driven spreading exhibit a small degree of non-normality, which explains the source of agreement between modal theory and experiment. The more rigorous generalized stability analysis presented here, however, affords critical insight into the types of disturbances leading to maximum unstable growth and the exact influence of the contact line model used. PMID- 12636604 TI - Growth and decay of localized disturbances on a surfactant-coated spreading film. AB - If the surface of a quiescent thin liquid film is suddenly coated by a patch of surface active material like a surfactant monolayer, the film is set in motion and begins spreading. An insoluble surfactant will rapidly attempt to coat the entire surface of the film thereby minimizing the liquid's surface tension. The shear stress that develops during the spreading process produces a maximum in surface velocity in the region where the moving film meets the quiescent layer. This region is characterized by a shock front with large interfacial curvature and a corresponding local buildup of surfactant which creates a spike in the concentration gradient. In this paper, we investigate the sensitivity of this region to infinitesimal disturbances. Accordingly, we introduce a measure of disturbance amplification and transient growth analogous to a kinetic energy that couples variations in film thickness to the surfactant concentration. These variables undergo significant amplification during the brief period in which they are convected past the downstream tip of the monolayer, where the variation in concentration gradient and surface curvature are largest. Once they migrate past this sensitive area, the perturbations weaken considerably and the system approaches a stable configuration. It appears that the localized disturbances of the type we consider here, cannot sustain asymptotic instability. Nonetheless, our study of the dynamics leading to the large transient growth clearly illustrates how the coupling of Marangoni and capillary forces work in unison to stabilize the spreading process against localized perturbations. PMID- 12636605 TI - Raman backscattering and amplification in a gas jet plasma. AB - Raman backscattering (RBS) of a picosecond 800-nm laser pulse from a gas jet plasma has been observed. The frequency shifts are in agreement with independent interferometric density measurements and the band width of the RBS is consistent with the linear growth rate. Raman amplification of a subpicosecond seed pulse, provided by an optical parametric oscillator and tuned to the spectral range of the RBS, has been obtained. The Raman resonance is confirmed by simultaneous plasma density measurements. PMID- 12636607 TI - Stability of certain families of ideal magnetohydrodynamic equilibria. AB - The equations of ideal magnetohydrodynamic equilibria posses a number of symmetries that may be used to generate a family of hitherto unknown equilibria if there exists a foliation of the original one by magnetic surfaces. In addition to the possibility of producing analytic equilibria from old ones, this family is studied to find among its members those with minimal energy, those lasting longer under slightly resistive conditions, and those linearly stable. It is shown that in general none of these properties implies any other, thus clarifying the difference among these concepts. PMID- 12636606 TI - Influence of optical thickness and hot electrons on Rydberg spectra of Ne-like and F-like copper ions. AB - Spectra in the 7.10 to 8.60 A range from highly charged copper ions are observed from three different laser-produced plasmas (LPPs). The LPPs are formed by a 15 ns Nd:glass laser pulse (type I: E(pulse)=1-8 J, lambda=1.064 microm), a 1-ps Nd:glass laser pulse (type II: E(pulse)=1 J, lambda=1.055 microm), and a 60-fs Ti:sapphire laser pulse (type III: E(pulse)=800 mJ, lambda=790 nm). The spectra of high-n (nor=5 x 10(10) W/cm(2)), phase explosion was found to occur after the completion of the laser pulse, but not during the process of laser energy deposition. This result is consistent with recent experiments. It is also found that the pressure of the induced ablation plasma plays a crucial role in the process of phase explosion. PMID- 12636615 TI - Mode spectra of thermally excited two-dimensional dust Coulomb clusters. AB - The mode spectra of finite Coulomb clusters, confined systems of charged microspheres in a complex plasma, have been determined experimentally. The spectral power density of all possible modes has been measured for particle numbers between N=3 and N=145 under different plasma conditions. The agreement between measured and calculated mode frequencies is found to be very good. From this, the parameters of the particle interaction, like particle charge and screening strength, have been extracted quantitatively. In addition, the particle and mode temperatures have been obtained. The modes and the particles show the same temperature and the principle of equipartitition holds for these systems. Moreover, certain modes of interest have been compared among the different clusters, as there are the breathing mode and intershell rotation as well as the lowest- and highest-frequency modes. Moreover, the mode-integrated spectrum shows two broad maxima which are explained from "shearlike" or "compressionlike" modes. From this analysis, the transition from finite number to crystal-like properties is observed to occur around N=12 particles. Finally, a model to visualize the transition from normal mode oscillations to wave dispersions in a 2D lattice has been proposed. PMID- 12636616 TI - Relativistic electron dynamics in intense crossed laser beams: acceleration and Compton harmonics. AB - Electron motion and harmonic generation are investigated in the crossed-beam laser-accelerator scheme in a vacuum. Exact solutions of the equations of motion of the electron in plane-wave fields are given, subject to a restricted set of initial conditions. The trajectory solutions corresponding to axial injection are used to calculate precise emission spectra. Guided by hindsight from the analytic investigations, numerical calculations are then performed employing a Gaussian beam representation of the fields in which terms of order epsilon(5), where epsilon is the diffraction angle, are retained. Present-day laser powers and initial conditions on the electron motion that simulate realistic laboratory conditions are used in the calculations. The analytic plane-wave work shows, and the numerical investigations confirm, that an optimal crossing angle exists, i.e., one that renders the electron energy gain a maximum for a particular set of parameters. Furthermore, the restriction to small crossing angles is not made anywhere. It is also shown that energy gains of a few GeV and energy gradients of several TeV/m may be obtained using petawatt power laser beams. PMID- 12636617 TI - Measurement of positron production efficiency from a tungsten monocrystalline target using 4- and 8-GeV electrons. AB - Intense positron sources are being widely investigated for the next-generation linear colliders and B factories. A new method utilizing an axially oriented crystal as a positron-production target is one of the bright schemes, since it provides a powerful photon source through channeling and coherent bremsstrahlung processes when high-energy electrons penetrate the target. A series of positron production experiments with tungsten crystals hit by 4- and 8-GeV single-bunch electron beams were carried out at the KEKB 8-GeV injector linac. Three tungsten crystals with different thicknesses (2.2, 5.3, and 9.0 mm) and those combined with amorphous tungsten plates were tested on a precise goniometer. The positron production yields were measured with a magnetic spectrometer in the positron momentum (P(e(+))) range from 5 to 20 MeV/c. The angle of the <111> crystal axis with respect to the electron-beam direction was controlled by measuring the relative intensities of the produced positrons as a function of the rotational angle of the goniometer. The results show that the enhancements of the positron yield from crystal targets compared to amorphous targets of the same thickness at P(e(+))=20 MeV/c are from 1.5 to 3.7 and from 1.8 to 5.1, depending upon the target thickness for 4- and 8-GeV electrons, respectively. PMID- 12636618 TI - Single-cycle electromagnetic pulses produced by oscillating electric dipoles. AB - We report an exact analytic solution of the Maxwell equations that is capable of describing single-cycle electromagnetic pulses beyond the slowly varying envelope approximation. The solution is based on the radiation field emitted by oscillating electric dipoles under the complex-source-point model. The spatiotemporal evolution of single-cycle electromagnetic pulses in free space is illustrated and discussed in detail by using the analytic solution obtained. PMID- 12636619 TI - Langevin description of speckle dynamics in nonlinear disordered media. AB - We formulate a Langevin description of dynamics of a speckle pattern resulting from the multiple scattering of a coherent wave in a nonlinear disordered medium. The speckle pattern exhibits instability with respect to periodic excitations at frequencies Omega below some Omega(max), provided that the nonlinearity exceeds some Omega-dependent threshold. A transition of the speckle pattern from a stationary state to the chaotic evolution is predicted upon increasing nonlinearity. The shortest typical time scale of chaotic intensity fluctuations is of the order of 1/Omega(max). PMID- 12636620 TI - Skyrmions in nematic liquid crystals. AB - Analytical solutions for static two-dimensional axisymmetric localized states minimizing the Frank free energy for nematic liquid crystals have been derived. These solitonic structures (skyrmions) include the well-known Belavin-Polyakov solutions as a special case for equal elastic constants. The structure and the equilibrium parameters of these nematic skyrmions crucially depend on values of the elastic constants. Stability limits of these structures and the possibility to observe them in nematic liquid crystals are discussed. PMID- 12636621 TI - Decay rate distributions of disordered slabs and application to random lasers. AB - We compute the distribution of the decay rates (also referred to as residues) of the eigenstates of a disordered slab from a numerical model. From the results of the numerical simulations, we are able to find simple analytical formulas which describe these results well. This is possible for samples both in the diffusive and in the localized regimes. As an example of a possible application, we investigate the lasing threshold of random lasers. PMID- 12636622 TI - High-order variational calculation for the frequency of time-periodic solutions. AB - We develop a convergent variational perturbation theory for the frequency of time periodic solutions of nonlinear dynamical systems. The power of the theory is illustrated by applying it to the Duffing oscillator. PMID- 12636623 TI - Underdamped commensurate dynamics in a driven Frenkel-Kontorova-type model. AB - The dynamics of a Frenkel-Kontorova chain subject to a substrate potential with a multiple-well structure and driven by an external dc force is studied in the underdamped regime. Making a rational choice among the three inherent length scales characterizing the system allows us to consider the possible formation of commensurate structures during sliding over the complex on-site potential. We comment both on the nature of the particle dynamics in the vicinity of the pinning-depinning transition point, and on the dynamical states displayed during the chain motion at different strengths of the dc driving. Varying the number of particles in the simulations allows us to consider, on a multiple-well substrate, the role played by the coverage variable on the depinning mechanism. The dependence of the minimal force required to initiate the chain motion (static friction) on the ratio of the model interaction strengths is analyzed and compared to the well-known case of the standard Frenkel-Kontorova model, which has only two inherent lengths. PMID- 12636624 TI - Energy exchange properties during second-harmonic generation in finite one dimensional photonic band-gap structures with deep gratings. AB - We study second-harmonic generation in finite, one-dimensional, photonic band-gap structures with large index contrast in the regime of pump depletion and global phase-matching conditions. We report a number of surprising results: above a certain input intensity, field dynamics resemble a multiwave mixing process, where backward and forward components compete for the available energy; the pump field is mostly reflected, revealing a type of optical limiting behavior; and second-harmonic generation becomes balanced in both directions, showing unusual saturation effects with increasing pump intensity. This dynamics was unexpected, and it is bound to influence the way one goes about thinking and designing nonlinear frequency conversion devices in a practical way. PMID- 12636625 TI - Discrete nonlinear Schrodinger equation with defects. AB - We investigate the dynamical properties of the one-dimensional discrete nonlinear Schrodinger equation (DNLS) with periodic boundary conditions and with an arbitrary distribution of on-site defects. We study the propagation of a traveling plane wave with momentum k: the dynamics in Fourier space mainly involves two localized states with momenta +/-k (corresponding to a transmitted and a reflected wave). Within a two-mode ansatz in Fourier space, the dynamics of the system maps on a nonrigid pendulum Hamiltonian. The several analytically predicted (and numerically confirmed) regimes include states with a vanishing time average of the rotational states (implying complete reflections and refocusing of the incident wave), oscillations around fixed points (corresponding to quasi-stationary states), and, above a critical value of the nonlinearity, self-trapped states (with the wave traveling almost undisturbed through the impurity). We generalize this treatment to the case of several traveling waves and time-dependent defects. The validity of the two-mode ansatz and the continuum limit of the DNLS are discussed. PMID- 12636626 TI - Stable vortex and dipole vector solitons in a saturable nonlinear medium. AB - We study both analytically and numerically the existence, uniqueness, and stability of vortex and dipole vector solitons in a saturable nonlinear medium in (2+1) dimensions. We construct perturbation series expansions for the vortex and dipole vector solitons near the bifurcation point, where the vortex and dipole components are small. We show that both solutions uniquely bifurcate from the same bifurcation point. We also prove that both vortex and dipole vector solitons are linearly stable in the neighborhood of the bifurcation point. Far from the bifurcation point, the family of vortex solitons becomes linearly unstable via oscillatory instabilities, while the family of dipole solitons remains stable in the entire domain of existence. In addition, we show that an unstable vortex soliton breaks up either into a rotating dipole soliton or into two rotating fundamental solitons. PMID- 12636627 TI - Multibarrier tunneling. AB - We study the tunneling through an arbitrary number of finite rectangular opaque barriers and generalize earlier results by showing that the total tunneling phase time depends neither on the barrier thickness nor on the interbarrier separation. We also predict peculiar features of the system considered, namely the independence of the transit time (for nonresonant tunneling) and the resonant frequency on the number of barriers crossed, which can be directly tested in photonic experiments. A thorough analysis of the role played by interbarrier multiple reflections and a physical interpretation of the results obtained is reported, showing that multibarrier tunneling is a highly nonlocal phenomenon. PMID- 12636628 TI - Thermal diffusion of supersonic solitons in an anharmonic chain of atoms. AB - We study the nonequilibrium diffusion dynamics of supersonic lattice solitons in a classical chain of atoms with nearest-neighbor interactions coupled to a heat bath. As a specific example we choose an interaction with cubic anharmonicity. The coupling between the system and a thermal bath with a given temperature is made by adding noise, delta correlated in time and space, and damping to the set of discrete equations of motion. Working in the continuum limit and changing to the sound velocity frame we derive a Korteweg-de Vries equation with noise and damping. We apply a collective coordinate approach which yields two stochastic ODEs which are solved approximately by a perturbation analysis. This finally yields analytical expressions for the variances of the soliton position and velocity. We perform Langevin dynamics simulations for the original discrete system which confirm the predictions of our analytical calculations, namely, noise-induced superdiffusive behavior which scales with the temperature and depends strongly on the initial soliton velocity. A normal diffusion behavior is observed for solitons with very low energy, where the noise-induced phonons also make a significant contribution to the soliton diffusion. PMID- 12636629 TI - Dynamics and stability of solitary waves in optical-microwave interaction. AB - We study the dynamics and the stability of localized bound states of optical and microwave fields, which are linked together by a quadratic nonlinearity. The system is an example of an intense interaction between low and high frequency waves, as appears in many areas of physics. Perturbed solitary waves show a number of regular but damped oscillations with strong radiation from the microwave. It is demonstrated that these oscillations are caused by the excitation of several quasibound asymmetric linear modes of the solitary wave. The associated eigenvalues are found to be complex leading to a decay of the oscillations as observed numerically. Additional quasibound linear modes with a complex eigenvalue corresponding to exponential growth also exist, but due to physical constraints cannot be excited. Therefore, in contrast to systems solely with high frequency waves, the stability of the solutions is retained. PMID- 12636630 TI - Optical properties of inverted opal photonic band gap crystals with stacking disorder. AB - We present a detailed study of the effect of planar stacking disorder on optical properties of inverted opal photonic crystals. Systems with periodic stacking sequences are first studied that include face centered cubic, hexagonal close packed, and doubly hexagonal close-packed photonic crystals. For the structures with periodic stacking order, we evaluate the band structure followed by calculation of transmission spectrum along the direction perpendicular to the hexagonal close-packing plane of the structures. Inverted opal photonic crystals with random stacking sequences are then studied by calculating average transmittance of the photonic crystal slabs over various random stacking configurations. The position and width of the lowest stop gap along the direction normal to the hexagonal close-packing plane is found to be invariant, regardless of the stacking sequence in the photonic crystal. We show how the propagation properties at higher frequencies are affected by the stacking configurations (both periodic and disordered stacking sequences), particularly those near the edges of the absolute band gap. The obtained results are directly relevant to transmission/reflection experiments on inverted opal photonic crystals with complete band gap. PMID- 12636631 TI - Exponentially fitted symplectic integrator. AB - In this paper a procedure for constructing efficient symplectic integrators for Hamiltonian problems is introduced. This procedure is based on the combination of the exponential fitting technique and symplecticness conditions. Based on this procedure, a simple modified Runge-Kutta-Nystrom second-order algebraic exponentially fitted method is developed. We give explicitly the symplecticness conditions for the modified Runge-Kutta-Nystrom method. We also give the exponential fitting and trigonometric fitting conditions. Numerical results indicate that the present method is much more efficient than the "classical" symplectic Runge-Kutta-Nystrom second-order algebraic method introduced by M.P. Calvo and J.M. Sanz-Serna [J. Sci. Comput. (USA) 14, 1237 (1993)]. We note that the present procedure is appropriate for all near-unimodal systems. PMID- 12636632 TI - Three-dimensional lattice-Boltzmann model of van der Waals fluids. AB - A three-dimensional lattice-Boltzmann model is developed for the simulation of nonideal fluids under static and flow conditions. The van der Waals formulation of quasilocal thermodynamics for nonuniform fluids is used, and the interfacial stress tensor for nonideal fluids appears explicitly in the hydrodynamic equations. The continuity and flow equations are fully recovered, and Galilean invariance is restored through appropriate manipulations of the pressure tensor. Although applied here to the D3Q15 lattice, the methodology of Galilean restoration can be easily modified for use with other three-dimensional lattices as well. The Laplace law and Gibbs-Thomson equations are satisfied with excellent accuracy by the model, as demonstrated by droplet equilibrium simulations. Spinodal decomposition and droplet coalescence simulations are also carried out, revealing a direct proportionality of the characteristic times to the viscosity, as expected. A wettability adjustment was made possible through the prescription of a chemical potential profile along the fluid-wall interface, and used for the simulation of droplet formation from a conical orifice. PMID- 12636633 TI - Betweenness centrality correlation in social networks. AB - Scale-free (SF) networks exhibiting a power-law degree distribution can be grouped into the assortative, dissortative, and neutral networks according to the behavior of the degree-degree correlation coefficient. Here we investigate the betweenness centrality (BC) correlation for each type of SF networks. While the BC-BC correlation coefficients behave similarly to the degree-degree correlation coefficients for the dissortative and neutral networks, the BC correlation is nontrivial for the assortative ones found mainly in social networks. The mean BC of neighbors of a vertex with BC g(i) is almost independent of g(i), implying that each person is surrounded by almost the same influential environments of people no matter how influential the person may be. PMID- 12636634 TI - Variational principle for stochastic wave and density equations. AB - We develop a stochastic generalization of the McLachlan variational principle and show that it can be used to derive known stochastic wave equations. We then use it to obtain an exact probability preserving stochastic density decomposition for vibrational dynamics problems with pairwise interaction. PMID- 12636635 TI - Critical temperature of the Potts models on the kagome lattice. AB - We investigate the Wu conjecture for the critical temperature of the Potts model on the kagome lattice. Previous testing of this conjecture has been for small values of q. Our emphasis is on large values of q where we are able to obtain very accurate estimates of the critical temperature which show the conjecture itself to be very accurate for these q values but not exact. PMID- 12636636 TI - Time-delay-induced stabilization of coupled discrete-time systems. AB - This paper shows that the time-delay-induced stabilization occurs in discrete time systems on numerical simulations. The stability analysis proves that this phenomenon never occurs in the discrete-time systems that have an odd-number property. This property is well known as the weak point of the delayed feedback control of chaos. Furthermore, we show that the phenomenon never occurs in any one-dimensional discrete-time system. PMID- 12636637 TI - Controlling fingering instabilities in rotating ferrofluids. AB - We perform a detailed analytic and numerical study of the evolution of a ferrofluid drop confined to a rotating Hele-Shaw cell in the presence of an azimuthal magnetic field. Our results demonstrate that the centrifugally driven interfacial instabilities can be simply controlled with the use of a current carrying wire. We compare an analytic linear analysis to our computational results and show that a number of observed features cannot be explained by linear theory alone, including a "diamond ring" instability that results when a droplet is nearly stabilized. PMID- 12636638 TI - Generation of a hollow ion beam: calculation of the rotation frequency required to accommodate symmetry constraint. AB - A hollow intense heavy ion beam with an annular focal spot has many important applications. The Gesellschaft fur Schwerionenforschung, Darmstadt is planning to develop a radio frequency wobbler that will rotate the beam at extremely high frequencies and thus create an annular (ring shaped) focal spot. In this paper, we present an analytical model that determines the minimum rotation frequency of the wobbler in order to achieve a high degree of irradiation symmetry (an asymmetry of a few percent) of the target. Estimates for a typical heavy ion imploded target are also presented. PMID- 12636639 TI - Density waves in traffic flow of two kinds of vehicles. AB - Through the car-following model, the traffic flow of two types of vehicles (cars and trucks) on a single-lane flow is studied, in which drivers on different vehicles have different sensitivities and the safety distance is assumed to be the same for all vehicles. The linear analysis is carried out to determine the condition of critical stability. With the nonlinear analysis, it proves that the small fluctuation of the vehicle density near the critical stable state satisfies the Korteweg-deVries equation and different sensitivities affect only the soliton evolution. When the headway in the critical state is more than the safety distance, the density around the soliton peak exceeds the density of the critical stable state, which can be explained as the formation of traffic jam. Contrarily, when the headway state is less than the safety distance, drivers will increase the headway to avoid the jam. The direct approach of the soliton perturbation shows that drivers' sensitivity will increase the soliton's amplitude continuously. Moreover, the increase of the number of trucks in the traffic flow will slow down the evolution of the amplitude. PMID- 12636640 TI - Interaction of a soliton with a continuous wave packet. AB - We analyze the scattering of a soliton from a continuous wave packet of arbitrary shape in a fiber, theoretically as well as numerically. Solitons recover their original shapes and velocities after collisions and the effect of collisions is described by the change of velocities of solitons. The theoretical predictions based on a WKB-type approximation are in a good agreement with numerical results. PMID- 12636641 TI - Simple approach for the bound-state energy spectrum of the generalized exponential-cosine Coulomb potential. AB - Based on the series expansion formalism, a relatively simple approach is proposed to solve the eigenvalues problems with partially screened and screened exponential-cosine Coulomb potentials. This approach is used to derive solutions to the Schrodinger equation with the two forms of potentials. The eigenenergies are explicitly deduced from solving the obtained corresponding polynomial equations. For illustration, high accuracy results have been obtained in the entire range of parameter values of these potential forms, with no constraints or adjustable constants. The present approach compares well, with existing methods, the results of which are precisely recovered as particular cases and does allow solutions to eigenvalues problems with any combination of potential parameters. PMID- 12636642 TI - Comment on "Power-law correlations in the southern-oscillation-index fluctuations characterizing El Nino". AB - In a recent publication [Phys. Rev. E 63, 047201 (2001)], Ausloos and Ivanova report power-law probability distributions, fractal properties, and antipersistent long-range correlations in the southern oscillation index. As a comparison with artificial short-range correlated data shows, most of these findings are possibly due to misleading interpretation of the analysis techniques used. PMID- 12636643 TI - Structure and phase diagram of high-density water: the role of interstitial molecules. AB - The structural transformations occurring to water from the low- to the high density regimes have been studied by classical molecular dynamics calculations. The local structure is analyzed through a proper choice of the relevant orientational distribution functions. This approach sheds light on the key role played by the interstitial molecules in the second coordination shell and identifies a clear structural fingerprint of high-density water. As a consequence, the analogy between the structure of high-density water and those of high-density ices is evidenced. PMID- 12636644 TI - Phase behavior and crystallization kinetics of poly-12-hydroxystearic-coated polymethylmethacrylate colloids. AB - Polymethylmethacrylate (PMMA) colloids sterically stabilized by a layer of chemically grafted poly-12-hydroxystearic (PHSA) are widely used in experiments as model hard spheres. However, due to the coating, the interaction between particles is slightly soft. Here we report a numerical study of the effect of the PHSA coating on the phase behavior and crystallization kinetics of PMMA colloids based on parameters determined from surface-force measurements on PHSA-PMMA coated mica surfaces [B. A. de L. Costello and P. F. Luckham, J. Colloid Interface Sci. 156, 72 (1993); B. A. de L. Costello et al., Langmuir 8, 464 (1992)]. We find that the core volume fraction of particles at freezing measured by Pusey and van Megen [Nature 320, 340 (1986)] can only be reproduced by using a thickness of the PHSA layer that is considerably larger than literature values. This may indicate that the particles are in fact slightly charged. Compared to perfect hard spheres, the crystallization rate in these slightly soft particles was found to be increased by about two orders of magnitudes. PMID- 12636645 TI - Two-stage surface freezing of a single top layer in a smectic-A membrane. AB - The crystallization of a single liquid top layer of smectic membranes of the compound 4O.8 has been studied with grazing-incidence x-ray diffraction. As this process takes place in two steps, involving an intermediate hexatic smectic-B layer before the final crystalline-B surface structure is reached, it provides a model for melting in two dimensions. The positional order has been investigated quantitatively by measuring the scattering profiles and the associated correlation lengths. The surface liquid-hexatic phase transition is found to be continuous, while the hexatic-crystal transition is weakly first order with an abrupt change of the in-plane positional correlations. The surface phase transitions do not modify the liquid in-plane structure of the interior layers. PMID- 12636646 TI - Field-induced switching between states of opposite chirality in a liquid crystalline phase. AB - A tilted smectic phase of a new achiral banana-shaped mesogen is presented. It possesses liquidlike order within the layers and appears with a fan-shaped texture. At sufficiently high electric fields, this texture can be transformed into a texture that displays a complete extinction between crossed polarizers and it forms randomly distributed chiral domains. Above a threshold these domains can be reversibly switched into a state of opposite handedness. PMID- 12636647 TI - Non-Hamiltonian equilibrium statistical mechanics. AB - In this paper the equilibrium statistical mechanics of non-Hamiltonian systems is formulated introducing an algebraic bracket. The latter defines non-Hamiltonian equations of motion in classical phase space according to the approach introduced in Phys. Rev. E 64, 056125 (2001). The Jacobi identity is no longer satisfied by the generalized bracket and as a result the algebra of phase space functions is not time translation invariant. The presence of a nonzero phase space compressibility spoils also the time-reversal invariance of the dynamics. The general Liouville equation is rederived and the properties of statistical averages are accounted for. The features of time correlation functions and linear response theory are also discussed. PMID- 12636648 TI - Stochastic resonance for nonlinear sensors with saturation. AB - We analyze the transmission of a noisy signal by sensor devices which are linear for small inputs and saturate at large inputs. Large information-carrying signals are thus distorted in their transmission. We demonstrate conditions where addition of noise to such large input signals can reduce the distortion that they undergo in the transmission. This is established for periodic, as well as aperiodic, and random information-carrying signals. Various measures characterizing the transmission, such as signal-to-noise ratio, input-output cross correlation, and mutual information, are shown improvable by addition of noise. These results constitute another instance of the nonlinear phenomenon of stochastic resonance where addition of noise enhances the signal. PMID- 12636649 TI - Some exact results for Boltzmann's annihilation dynamics. AB - The problem of ballistic annihilation for a spatially homogeneous system is revisited within Boltzmann's kinetic theory in two and three dimensions. Analytical results are derived for the time evolution of the particle density for some isotropic discrete bimodal velocity modulus distributions. According to the allowed values of the velocity modulus, different behaviors are obtained: power law decay with nonuniversal exponents depending continuously upon the ratio of the two velocities, or exponential decay. When one of the two velocities is equal to zero, the model describes the problem of ballistic annihilation in the presence of static traps. The analytical predictions are shown to be in agreement with the results of two-dimensional molecular dynamics simulations. PMID- 12636650 TI - Stochastic resonance driven by two different kinds of colored noise in a bistable system. AB - The phenomenon of stochastic resonance in a bistable nonlinear system is investigated when both the multiplicative noise and the coupling between additive and multiplicative noise are colored with different values of noise correlation time tau(1) and tau(2). Combining the functional analysis and unified colored noise approximation, the two different kinds of colored noise in the nonlinear system can be simplified. The signal-to-noise ratio is calculated when a weakly periodic signal is added to the system. It is found that there appears a transition between one peak and two peaks in the curve of the signal-to-noise ratio when either the noise correlation time tau(1) and tau(2) or the coupling strength lambda between additive and multiplicative noise is increased. The transition between one and two peaks depending on tau(1) and lambda is more complex than that depending on tau(2). PMID- 12636651 TI - Homogeneous shear flow of a hard-sphere fluid: analytic solutions. AB - Recently, a solution for collision-free trajectories in an N particle thermostatted hard-sphere system undergoing homogeneous shear (the so-called "Sllod" equations of motion) led to a kinetic theory of dilute hard-sphere gases under shear. However, a solution for collisions, necessary for a complete theory at higher densities, has been missing. We present an analytic solution to this problem, which provides surprising insights into the mechanical aspects of thermostatting a system in an external field. The equivalence of constant temperature and constant energy ensembles in the thermodynamic limit in equilibrium, the conditions for the nature of heat exchange with the environment (entropy creation and reduction) in the system, and the condition for appearance of the artificial string phase follow from our solution. PMID- 12636652 TI - Application of minimal subtraction renormalization to crossover behavior near the 3He liquid-vapor critical point. AB - Parametric expressions are used to calculate the isothermal susceptibility, specific heat, order parameter, and correlation length along the critical isochore and coexistence curve from the asymptotic region to crossover region. These expressions are based on the minimal-subtraction renormalization scheme within the phi(4) model. Using two adjustable parameters in these expressions, we fit the theory globally to recently obtained experimental measurements of isothermal susceptibility and specific heat along the critical isochore and coexistence curve, and early measurements of coexistence curve and light scattering intensity along the critical isochore of 3He near its liquid-vapor critical point. The theory provides good agreement with these experimental measurements within the reduced temperature range |t|>1], in two and in three dimensions. The other two involve long-ranged repulsive forces in two (dipolar, r(-3) potential) and in three (screened Coulomb, or repulsive Yukawa interactions) dimensions. We find that the theory, without any sort of adjustable parameters or rescaling prescriptions, provides an excellent approximate description of the collective dynamics of these model systems, particularly in the short- and intermediate-time regimes. We also compare our results with those of the single exponential approximation and with the competing mode-mode coupling theory. PMID- 12636655 TI - Long-range correlations in the electric signals that precede rupture: further investigations. AB - The correlations within the time series of the seismic electric signal (SES) activities have been studied in a previous paper [P. Varotsos, N. Sarlis, and E. Skordas, Phys. Rev. E 66, 011902 (2002)]. Here, we analyze the time series of successive high- and low-level states' durations. The existence of correlation between the states is investigated by means of Hurst and detrended fluctuation analysis (DFA). The multifractal DFA (MF-DFA) is also employed. The results point to a stronger correlation, and hence longer memory, in the series of the high level states. Furthermore, an analysis in the "natural" time domain reveals that certain power spectrum characteristics seem to distinguish SES activities from "artificial" (man-made) electric noises. More precisely, for natural frequencies 0 of infinite horizon billiards, given a bounded initial distribution of particles. For a variety of billiard models we find <|r|(q)> approximately t(gamma(q)) (up to factors of ln t). The time exponent, gamma(q), is piecewise linear and equal to q/2 for q<2 and q-1 for q>2. We discuss the lack of dependence of this result on the initial distribution of particles and resolve apparent discrepancies between this time dependence and a prior result. The lack of dependence on initial distribution follows from a remarkable scaling result that we obtain for the time evolution of the distribution function of the angle of a particle's velocity vector. PMID- 12636657 TI - Fractional dynamics from the ordinary Langevin equation. AB - We consider the usual Langevin equation depending on an internal time. This parameter is substituted by a first passage time of a self-similar Markov process. Then the Gaussian process is parent, and the hitting time process is directing. The probability to find the resulting process at the real time is defined by the integral relationship between the probability densities of the parent and directing processes. The corresponding master equation becomes the fractional Fokker-Planck equation. We show that the resulting process has non Markovian properties, all its moments are finite, the fluctuation-dissipation relation and the H-theorem hold. PMID- 12636659 TI - Noise-driven mechanism for pattern formation. AB - We extend the mechanism for noise-induced phase transitions proposed by Ibanes et al. [Phys. Rev. Lett. 87, 020601 (2001)] to pattern formation phenomena. In contrast with known mechanisms for pure noise-induced pattern formation, this mechanism is not driven by a short-time instability amplified by collective effects. The phenomenon is analyzed by means of a modulated mean field approximation and numerical simulations. PMID- 12636658 TI - Continuous-time random-walk model for financial distributions. AB - We apply the formalism of the continuous-time random walk to the study of financial data. The entire distribution of prices can be obtained once two auxiliary densities are known. These are the probability densities for the pausing time between successive jumps and the corresponding probability density for the magnitude of a jump. We have applied the formalism to data on the U.S. dollar-deutsche mark future exchange, finding good agreement between theory and the observed data. PMID- 12636660 TI - Symmetric linear kinetic theory. AB - Equilibrium time correlation functions are expressed by two Onsager-symmetric quantumlike operators containing equilibrium distribution function. A model of dissipation for smooth interaction potentials is proposed. Approximations leading to the mode coupling formula are clearly stated and the Green-Kubo prescription for the calculation of the transport coefficients is reformulated. PMID- 12636661 TI - Local density approach for modeling fluids with density-dependent interactions. AB - In a recent paper [Phys. Rev. Lett. 86, 2038 (2001)] a simple fluid with a particular density-dependent pair potential was shown to exhibit, together with the vapor-liquid transition, a liquid-liquid phase separation and it was evidenced that, in order to adequately define the correct boundaries of stability, a simulation procedure based on the use of local densities had to be devised. It was found that for certain thermodynamic states the potential drives the system toward a phase separation that is otherwise frustrated by the change in the interactions induced by density fluctuations. Therefore, when integral equations or global density simulations are used, the critical points estimated from the thermodynamics are not associated with divergent correlations and vice versa. Here, we will explore in depth this fluid and introduce a detailed account of the proposed local density simulation technique. The results presented bear general significance for density-dependent potentials, like those of liquid metals or charge-stabilized colloids. PMID- 12636662 TI - Theory of the lattice Boltzmann method: three-dimensional model for linear viscoelastic fluids. AB - A three-dimensional lattice Boltzmann model with thirty two discrete velocity distribution functions for viscoelastic fluid is presented in this work. The model is based upon the generalized lattice Boltzmann equation constructed in moment space. The nonlinear equilibria of the model have a number of coupling constants that are free parameters. The dispersion equation of the model is analyzed under various conditions to obtain the constraints on the free parameters such that the model satisfies isotropy and Galilean invariance. The macroscopic equations are also derived from the lattice Boltzmann model through the dispersion equation analysis and the Chapman-Enskog analysis. We demonstrate that the dispersion equation analysis can be used as a general and effective means to derive hydrodynamic equations, excluding some nonlinear source terms, from the lattice Boltzmann model, to obtain conditions for its isotropy and Galilean invariance, and to optimize its stability. We show that the hydrodynamic behavior of the lattice Boltzmann model has memory effects, and that in the linear regime, it behaves as a viscoelastic fluid described by the Jeffreys model. Some numerical results to verify the theoretical analysis of the model are also presented. PMID- 12636663 TI - Heat conductivity in linear mixing systems. AB - We present analytical and numerical results on the heat conduction in a linear mixing system. In particular we consider a quasi-one-dimensional channel with triangular scatterers with internal angles which are irrational multiples of pi, and we show that the system obeys the Fourier law of heat conduction. Therefore, deterministic diffusion and normal heat transport which are usually associated with full hyperbolicity, actually take place in systems without exponential instability. PMID- 12636664 TI - Bulk viscosity in the case of the interatomic potential depending on density. AB - We derive a formula for the bulk viscosity zeta in a density-dependent-potential system. This is a generalization of the bulk-viscosity formula for the density independent-potential system which has been proposed recently by us. In our formulas, the bulk viscosity is expressed by using microscopic quantities such as interatomic potentials and pair distribution functions. This has an outstanding advantage of providing the relation between such microscopic information and a macroscopic quantity zeta. On the other hand, in all formulas proposed previously, zeta is expressed in terms of pressure, a macroscopic quantity, and it is difficult to discuss this relation. We apply our formula to a model liquid metal in which the interatomic potential varies with density. Our calculated results show that zeta increases in the density region where the interatomic potential changes from one type to another. These results agree qualitatively with the experimental results about liquid mercury. PMID- 12636665 TI - Hard-sphere limit of soft-sphere model for granular materials: stiffness dependence of steady granular flow. AB - Dynamical behavior of steady granular flow is investigated numerically in the inelastic hard-sphere limit of the soft-sphere model. We find distinctively different limiting behaviors for the two flow regimes, i.e., the collisional flow and the frictional flow. In the collisional flow, the hard-sphere limit is straightforward; the number of collisions per particle per unit time converges to a finite value and the total contact time fraction with other particles goes to zero. For the frictional flow, however, we demonstrate that the collision rate diverges as the power of the particle stiffness so that the time fraction of the multiple contacts remains finite even in the hard-sphere limit, although the contact time fraction for the binary collisions tends to zero. PMID- 12636666 TI - Clustering transitions in vibrofluidized magnetized granular materials. AB - We study the effects of long-range interactions on the phases observed in cohesive granular materials. At high vibration amplitudes, a gas of magnetized particles is observed with velocity distributions similar to nonmagnetized particles. Below a transition temperature compact clusters are observed to form and coexist with single particles. The cluster growth rate is consistent with a classical nucleation process. However, the temperature of the particles in the clusters is significantly lower than the surrounding gas, indicating a breakdown of equipartition. If the system is quenched to low temperatures, a metastable network of connected chains self-assemble due to the anisotropic nature of magnetic interactions between particles. PMID- 12636667 TI - Shearing of loose granular materials: a statistical mesoscopic model. AB - A two-dimensional lattice model for the formation and evolution of shear bands in granular media is proposed. Each lattice site is assigned a random variable which reflects the local density. At every time step, the strain is localized along a single shear band which is a spanning path on the lattice chosen through an extremum condition. The dynamics consists of randomly changing the "density" of the sites only along the shear band, and then repeating the procedure of locating the extremal path and changing it. Starting from an initially uncorrelated density field, it is found that this dynamics leads to a slow compaction along with a nontrivial patterning of the system, with high-density regions forming which shelter long-lived low-density valleys. Further, as a result of these large density fluctuations, the shear band, which was initially equally likely to be found anywhere on the lattice, gets progressively trapped for longer and longer periods of time. This state is, however, metastable, and the system continues to evolve slowly in a manner reminiscent of glassy dynamics. Several quantities have been studied numerically which support this picture and elucidate the unusual system-size effects involved. PMID- 12636668 TI - Shock waves in two-dimensional granular flow: effects of rough walls and polydispersity. AB - We have studied the two-dimensional flow of balls in a small-angle funnel, when either the side walls are rough or the balls are polydisperse. As in earlier work on monodisperse flows in smooth funnels, we observe the formation of kinematic shock waves (density waves). We find that for rough walls the flows are more disordered than for smooth walls and that shock waves generally propagate more slowly. For rough wall funnel flow, we show that the shock velocity and frequency obey simple scaling laws. These scaling laws are consistent with those found for smooth wall flow, but here they are cleaner since there are fewer packing-site effects and we study a wider range of parameters. For pipe flow (parallel side walls), rough walls support many shock waves, while smooth walls exhibit fewer or no shock waves. For funnel flows of balls with varying sizes, we find that flows with weak polydispersity behave qualitatively similar to monodisperse flows. For strong polydispersity, scaling breaks down and the shock waves consist of extended areas where the funnel is blocked completely. PMID- 12636669 TI - Model of coarsening and vortex formation in vibrated granular rods. AB - Neicu et al. observed experimentally spontaneous formation of the long-range orientational order and large-scale vortices in a system of vibrated macroscopic rods. We propose a phenomenological theory of this phenomenon based on a coupled system of equations for local rods density and tilt. The density evolution is described by the modified Cahn-Hilliard equation, while the tilt is described by the Ginzburg-Landau type equation. Our analysis shows that, in accordance with the Cahn-Hilliard dynamics, islands of the ordered phase appear spontaneously and grow due to coarsening. The generic vortex solutions of the Ginzburg-Landau equation for the tilt correspond to the vortical motion of the rods around the cores which are located near the centers of the islands. PMID- 12636670 TI - Onset of thermal convection in a horizontal layer of granular gas. AB - The Navier-Stokes granular hydrodynamics is employed for determining the threshold of thermal convection in an infinite horizontal layer of granular gas. The dependence of the convection threshold, in terms of the inelasticity of particle collisions, on the Froude and Knudsen numbers is found. A simple necessary condition for convection is formulated in terms of the Schwarzschild's criterion, well known in thermal convection of (compressible) classical fluids. The morphology of convection cells at the onset is determined. At large Froude numbers, the Froude number drops out of the problem. As the Froude number goes to zero, the convection instability turns into a recently discovered phase separation instability. PMID- 12636671 TI - Velocity distribution of fluidized granular gases in the presence of gravity. AB - The velocity distribution of a fluidized dilute granular gas in the direction perpendicular to the gravitational field is investigated by means of molecular dynamics simulations. The results indicate that the velocity distribution can be exactly described neither by a Gaussian nor by a stretched exponential law. Moreover, it does not exhibit any kind of scaling. In fact, the actual shape of the distribution depends on the number of monolayers at rest, on the restitution coefficient and on the height at what it is measured. The role played by the number of particle-particle collisions as compared with the number of particle wall collisions is discussed. PMID- 12636672 TI - Shear viscosity for a heated granular binary mixture at low density. AB - The shear viscosity for a heated granular binary mixture of smooth hard spheres at low density is analyzed. The mixture is heated by the action of an external driving force (Gaussian thermostat) that exactly compensates for cooling effects associated with the dissipation of collisions. The study is made from the Boltzmann kinetic theory, which is solved by using two complementary approaches. First, a normal solution of the Boltzmann equation via the Chapman-Enskog method is obtained up to first order in the spatial gradients. The mass, heat, and momentum fluxes are determined and the corresponding transport coefficients identified. As in the free cooling case [V. Garzo and J. W. Dufty, Phys. Fluids 14, 1476 (2002)], practical evaluation requires a Sonine polynomial approximation, and here it is mainly illustrated in the case of the shear viscosity. Second, to check the accuracy of the Chapman-Enskog results, the Boltzmann equation is numerically solved by means of the direct simulation Monte Carlo method. The simulation is performed for a system under uniform shear flow, using the Gaussian thermostat to control inelastic cooling. The comparison shows an excellent agreement between theory and simulation over a wide range of values of the restitution coefficients and the parameters of the mixture (masses, concentrations, and sizes). PMID- 12636673 TI - Diffusion-controlled growth: theory and closure approximations. AB - We expand upon a new theoretical framework for diffusion-limited aggregation and associated dielectric breakdown models in two dimensions [R. C. Ball and E. Somfai, Phys. Rev. Lett. 89, 135503 (2002)]. Key steps are understanding how these models interrelate when the ultraviolet cut-off strategy is changed, the analogy with turbulence, and the use of logarithmic field variables. Within the simplest, Gaussian, truncation of mode-mode coupling, all properties can be calculated. The agreement with prior knowledge from simulations is encouraging, and a new superuniversality of the tip scaling exponent is discussed. We find angular resonances relatable to the cone angle theory, and we are led to predict a new screening transition in the DBM at large eta. PMID- 12636674 TI - Field-induced structures in miscible ferrofluid suspensions with and without latex spheres. AB - We explore magnetic-field-induced ordering and microphase separation of aqueous ferrofluid and of aqueous mixtures of ferrofluid with nonmagnetic latex spheres. The ferrofluid is a surfactant stabilized aqueous suspension of magnetite (Fe3O4) particles with average diameter 20 nm (including the approximately 2.5-nm thick surfactant layer); the nonmagnetic latex spheres are charge stabilized polymethylmethacrylate (PMMA) particles with diameters of 42 nm, 108 nm, and 220 nm. In the presence of a uniform magnetic field, needlelike ferrofluid droplets formed that eventually grew to sample-traversing columns at fields of approximately 600 G; the two-dimensional structure of these columns was, however, glassy rather than hexagonal. In higher fields, approximately 1000 G, the columns stretched and coalesced into sheetlike striped liquids, but a true lamellar phase was not observed. The addition of nonmagnetic latex spheres to the ferrofluid suspension lowered substantially the critical field for the formation of columns, and induced lamellar (stripe) phases at relatively low applied fields. Image analysis was used to determine the spatial correlation functions, the average needle or column spacing, and the average lamellae spacing of these samples as a function of latex sphere size and concentration. PMID- 12636675 TI - Dielectrophoresis of charged colloidal suspensions. AB - We present a theoretical study of dielectrophoretic (DEP) crossover spectrum of two polarizable particles under the action of a nonuniform ac electric field. For two approaching particles, the mutual polarization interaction yields a change in their respective dipole moments, and hence, in the DEP crossover spectrum. The induced polarization effects are captured by the multiple image method. Using spectral representation theory, an analytic expression for the DEP force is derived. We find that the mutual polarization effects can change the crossover frequency at which the DEP force changes sign. The results are found to be in agreement with recent experimental observation and as they go beyond the standard theory, they help to clarify the important question of the underlying polarization mechanisms. PMID- 12636676 TI - Phase-separation-induced fractionation in molar mass in aqueous mixtures of gelatin and dextran. AB - An overview of the effects of phase separation of aqueous mixtures of gelatin and dextran on the fractionation in molar mass of these two components is given. Molar mass distributions in coexisting phases were investigated using size exclusion chromatography with multiangle laser light scattering. The initial molar mass of the native material, concentration, and temperature were varied. The results show a strong fractionation in molar mass for both components. The molar mass of the native material and concentration appeared to be the only factors that affected the final molar mass distributions, temperature having no effect. The results show that in the molar mass range where fractionation is the strongest, i.e., roughly below the maximum in the distribution, fractionation is governed by a Boltzmann factor e(-deltaG/kT), where deltaG denotes the free energy involved in transferring a polymer with a certain length from the enriched to the depleted phase, and in this case turns out to be proportional to the molar mass. Comparison of the results of phase separation with results on dialysis shows that water affinity is not the driving force for the phase separation of gelatin and dextran in aqueous solution. The gelation properties of gelatin in both phases were also determined. The gelation properties of gelatin in the coexisting phases differ from those of native gelatin. In particular, the gelatin in the gelatin-poor phase shows strong differences compared to the native material. PMID- 12636677 TI - Dynamics of yielding observed in a three-dimensional aqueous dry foam. AB - We study the onset of yielding in stable three-dimensional dry foams following the start up of steady shear flow. By means of a charge-coupled device camera equipped with a small depth-of-field objective, we visualize the Plateau border network in the bulk of the foam. The onset of yielding is identified with the deformation gamma(c) for which shear induced rearrangements start occurring. We show that gamma(c) is independent of shear rate gamma; in a quasistatic regime whereas at high strain rates, a rapid increase of gamma(c) with gamma; is observed, in qualitative agreement with theoretical models. Moreover, spatiotemporal image analyses are used to determine the velocity profile in the gap. We find that this profile remains linear up to strains far beyond gamma(c). Moreover, we have studied the strain history dependence of gamma(c). PMID- 12636678 TI - Dynamics of supercooled liquids confined to the pores of sol-gel glass: a dynamic light scattering study. AB - Dynamics of low molecular weight and polymeric glass forming liquids in confined geometries has been studied by means of depolarized dynamic light scattering: photon correlation spectroscopy and Fabry-Perot interferometry. The pore size of the glassy matrix amounted to 2.5, 5.0, and 7.5 nm. The glass transition temperature T(g) of these liquids in confined geometries has been measured using differential scanning calorimetry. A systematic decrease of T(g) (up to 25 K) with decreasing pore size has been observed. The relaxation times of the alpha process at constant temperature were decreasing with decreasing pore size (up to 6 orders of magnitude at T(g)), while the width of their distribution was increasing. The change of the relaxation times can be assigned to the change of T(g) in confined geometries. After correcting the activation plots for the shift of T(g) a master curve was obtained for all pore sizes and the bulk material. The effect of chemical modification of the surface of the porous matrix on the dynamics of ortho-terphenyl has also been studied. These dramatic changes of the T(g) and the relaxation time of confined liquids can be explained by simple thermodynamic arguments. There is no indication that they are related to the change of the correlation length of cooperative dynamics. PMID- 12636679 TI - Effect of composition changes on the structural relaxation of a binary mixture. AB - Within the mode-coupling theory for idealized glass transitions, we study the evolution of structural relaxation in binary mixtures of hard spheres with size ratios delta of the two components varying between 0.5 and 1.0. We find two scenarios for the glassy dynamics. For small size disparity, the mixing yields a slight extension of the glass regime. For larger size disparity, a plasticization effect is obtained, leading to stabilization of the liquid due to mixing. For all delta, a decrease of the elastic moduli at the transition due to mixing is predicted. A stiffening of the glass structure is found as is reflected by the increase of the Debye-Waller factors at the transition points. The critical amplitudes for density fluctuations at small and intermediate wave vectors decrease upon mixing, and thus the universal formulas for the relaxation near the plateau values describe a slowing down of the dynamics upon mixing for the first step of the two-step relaxation scenario. The results explain the qualitative features of mixing effects reported by Williams and van Megen [Phys. Rev. E 64, 041502 (2001)] for dynamical light-scattering measurements on binary mixtures of hard-sphere-like colloids with size ratio delta=0.6. PMID- 12636680 TI - Connectivity strategies to enhance the capacity of weight-bearing networks. AB - The connectivity properties of a weight-bearing network are exploited to enhance its capacity. We study a 2D network of sites where the weight-bearing capacity of a given site depends on the capacities of the sites connected to it in the layers above. The network consists of clusters, viz., a set of sites connected with each other with the largest such collection of sites being denoted as the maximal cluster. New connections are made between sites in successive layers using two distinct strategies. The key element of our strategies consists of adding as many disjoint clusters as possible to the sites on the trunk T of the maximal cluster. In the first strategy the reconnections start from the last layer upwards and stop when no new sites are added. In the second case, the reconnections start from the top layer and go all the way down to the last layer. The new networks can bear much higher weights than the original networks and have much lower failure rates. The first strategy leads to a greater enhancement of stability, whereas the second leads to a greater enhancement of capacity compared to the original networks. The original network used here is a typical example of the branching hierarchical class. However, the application of strategies similar to ours can yield useful results in other types of networks as well. PMID- 12636681 TI - Scattering from dilute ferrofluid suspensions in soft polymer gels. AB - Small angle neutron and x-ray scattering methods are used to investigate the structure of dilute suspensions of two different ferrofluid systems dispersed in soft polyacrylamide hydrogels. It is found that the particles in the fluid are fractal aggregates composed of smaller particles of radius ca. 5 nm. The fractal dimension is strongly dependent on sample, taking the value 1.7 in the first sample and 2.9 in the second sample. In the presence of a magnetic field the aggregates orient, but are restricted in both their translational and rotational freedom. The effect of the gel elasticity is treated as a hindrance to the orientation process. PMID- 12636682 TI - Experimental studies of the ortho-toluidine glass transition. AB - The thermodynamic and dynamics proprieties of ortho-toluidine, in the vicinity of a glass transition, have been studied by calorimetric and by two light scattering techniques, depolarized light scattering and time-resolved optical Kerr effect. Differential scanning microcalorimetry clearly detects a glass transition in o toluidine and it measures some thermodynamics critical parameters, in particular, the transition temperature. The light scattering data have been analyzed according to the mode-coupling theory. This theory gives a good interpretation of our data and it allows to extract safely the critical parameters of the o toluidine dynamics. We found a fair agreement between the analysis outputs performed in the frequency domain and in the time domain. Finally, we compared the glass transition features of o-toluidine with that of its isomer meta toluidine, looking for some general idea about the molecular aspects of the glass transition. PMID- 12636684 TI - Binary mixtures of magnetic fluids. AB - We study a binary mixture of a van der Waals fluid and a ferromagnetic fluid at zero magnetic field on the basis of the mean field Ising fluid model and the van der Waals theory with quadratic mixing rules. Depending on three reduced parameters, the phase diagram shows a surface of magnetic phase transitions and lines of tricritical points, critical end points, and magnetic consolute points. First-order phase transition surfaces and critical lines are calculated numerically. For the line of tricritical points, which can occur in two different topologies, an analytic expression is derived. All higher-order lines and coexistence surfaces are visualized in three-dimensional x, T, p and xi, T, p diagrams, where xi is a mapping of delta, the conjugated field of the mole fraction x, on the unit interval. PMID- 12636683 TI - Probes of heterogeneity in rotational dynamics: application to supercooled liquid CS2. AB - The distribution of individual molecular contributions to the second-rank rotational correlation function is introduced and used to construct probes of heterogeneity in rotational dynamics. The ideas are tested in a molecular dynamics simulation of supercooled liquid CS2. Both the quantity of heterogeneity and its lifetime or exchange time tau(ex) increase as the temperature is lowered through the supercooled state, and increase strongly as the mode-coupling temperature T(c) is approached. Crossover from Arrhenius to super-Arrhenius behavior of the rotational relaxation times tau(1) and tau(2) is observed, direct evidence of fragility in CS2. The T dependence of tau(ex) is stronger than that of the rotational times, and it may approach them from below at T(c), although the simulation is then very difficult. A detailed characterization of other aspects of the dynamical crossover is obtained, and the general implications of rotational heterogeneity for supercooled dynamics are discussed. PMID- 12636685 TI - Cohen-Grest model for the dynamics of supercooled liquids. AB - Recent experiments have established that, at least for van der Waals glass formers, volume fluctuations contribute significantly to the slowing down of the dynamics near T(g). Accordingly, we use the Cohen-Grest (CG) free-volume model to analyze dielectric relaxation data for six van der Waals liquids. The CG equation accurately describes the structural relaxation times over broader ranges of temperature than the more common Vogel-Fulcher relation. Moreover, the CG equation requires two less adjustable parameters when the data span the Stickel temperature T(B) associated with a change in the dynamics. The characteristic temperature T0 of the CG model can be identified with T(B), suggesting that the crossover reflects onset of percolation of the free volume. The CG parameters used to fit the structural relaxation times allow the free volume per liquidlike molecule to be calculated. These results, however, are at odds with free-volume estimates extracted from pressure-volume-temperature data. PMID- 12636686 TI - Grazing-incidence x-ray diffraction study of Langmuir films of amphiphilic monodendrons. AB - We have used pressure-area isotherms and grazing-incidence x-ray diffraction to study structures of Langmuir films of first-generation monodendrons with two or three peripheral alkyl chains. Unlike the structures observed in their bulk liquid crystalline mesophases, these multichain monodendrons form either a centered rectangular lattice with molecular axes tilted toward nearest neighbors or an oblique lattice with molecular axes tilted in low-symmetry directions. PMID- 12636687 TI - Depinning of elastic manifolds. AB - We compute roughness exponents of elastic d-dimensional manifolds in (d+1) dimensional embedding spaces at the depinning transition for d=1, em leader,4. Our numerical method is rigorously based on a Hamiltonian formulation; it allows us to determine the critical manifold in finite samples for an arbitrary convex elastic energy. For a harmonic elastic energy (Delta(2) model), we find values of the roughness exponent between the one-loop and two-loop functional renormalization group results, in good agreement with earlier cellular automaton simulations. We find that the Delta(2) model is unstable with respect both to slight stiffening and to weakening of the elastic potential. Anharmonic corrections to the elastic energy allow us to obtain the critical exponents of the quenched Kardar, Parisi, Zhang class. PMID- 12636688 TI - Depinning with dynamic stress overshoots: a hybrid of critical and pseudohysteretic behavior. AB - A model of an elastic manifold driven through a random medium by an applied force F is introduced and studied. The focus is on the effects of inertia and elastic waves, in particular stress overshoots in which motion of one segment of the manifold causes a temporary stress on its neighboring segments in addition to the static stress. Such stress overshoots decrease the critical force for depinning and make the depinning transition hysteretic with static and pinned configurations coexisting with the steadily moving phase for a range of F. We find that the steady-state velocity of the moving phase is, nevertheless, history independent and the critical behavior as the force is decreased is in the same universality class as in the absence of stress overshoots-the dissipative limit in which hysteresis cannot occur and theoretical analysis has been possible. To reach this conclusion, finite-size scaling analyses have been performed and a variety of quantities studied, including velocities, roughnesses, distributions of critical forces, and universal amplitude ratios. If the force is increased slowly from zero, the behavior is complicated with a spectrum of avalanche sizes occurring that seems to be quite different from the dissipative limit. Related behavior is seen as the force is increased back up again to restart the motion of samples that have been stopped from the moving phase. The restarting process itself involves both fractal-like and bubblelike nucleation. Hysteresis loops in small- and intermediate-size samples can be understood in terms of a depletion layer caused by the stress overshoots. Surprisingly, in the limit of very large samples the hysteresis loops vanish. Although complicated crossovers complicate the analysis, we argue that the underlying universality class governing this pseudohysteresis and avalanches is again that of the apparently very different dissipative limit. But there are history dependent amplitudes-associated with the depletion layer-that cause striking differences over wide ranges of length scales. Consequences of this picture for the statistics and dynamics of earthquakes on geological faults are briefly discussed. PMID- 12636689 TI - Influence of surface roughness on the adhesion of elastic films. AB - It is shown that a self-affine roughness at the junction of an elastic film and a hard solid substrate influences considerably the adhesion of the elastic film, especially for small roughness exponents H (H<0.5) and/or large long wavelength roughness ratios w/xi with w being the rms roughness amplitude and xi being the in-plane roughness correlation length. Analytical calculations of the local surface slope allows an estimate of the roughness effects on the adhesion energy more precisely than those presented in earlier works (especially for roughness exponents H<0.5). For weak surface roughness the elastic energy contribution is significant on the film effective surface energy deltagamma(eff) and on pull-off force for elastic modulus E in the range of GPa. Moreover, in the case of partial contact an estimation of the pull-off force shows that it strongly decreases with reducing contact area due to surface. PMID- 12636690 TI - Nucleation probability in binary heterogeneous nucleation of water-n-propanol vapor mixtures on insoluble and soluble nanoparticles. AB - Nucleation probabilities for binary heterogeneous nucleation have been measured quantitatively. Heterogeneous nucleation of binary n-propanol-water vapor mixtures on partially soluble 6.9-nm NaCl particles and on nonsoluble oxidized and nonoxidized 8-nm Ag particles at a constant nucleation temperature of 288 K (NaCl) or 285 K (Ag) has been investigated experimentally and theoretically. An expansion chamber was applied to generate supersaturated vapor mixtures. Number concentrations of particles activated to condensational growth were determined optically for various vapor phase activities at constant temperature. Nucleation probabilities have been measured as functions of the vapor phase activities for several n-propanol-water vapor mixing ratios. Beyond certain vapor phase activities a sharp onset of heterogeneous nucleation was observed both for NaCl and for Ag particles. For insoluble particle surfaces the theoretical slopes of the nucleation probability versus vapor phase activity curves were found to be somewhat steeper as compared to the experiment. On the other hand, for soluble particles the experimental slopes were found to be in satisfactory agreement with theory. The onset of the nucleation process is defined at an activity value where 50% of the particles were activated to condensational growth. Onset activities were obtained for various mixing ratios of the binary vapor mixtures. In the case of NaCl nuclei for n-propanol rich mixtures, the heterogeneous nucleation calculations based on a thermodynamically consistent version of Fletcher theory and an experimentally determined contact angle provide a reasonable approximation of the experimental data. However, it appears that the Fletcher theory is not applicable in the region of transition from n-propanol rich to water rich mixtures. Based on the Kohler theory of activation of soluble particles, a theory was formulated accounting for the presence of two condensable vapors and limited solubility of NaCl in water-propanol liquid mixtures. This approach provides a satisfactory description of the transition from activation of soluble particles to nucleation of vapors on insoluble particles. In the case of silver particles Fletcher theory with macroscopic experimental contact angles as well as with contact angle zero results in strong deviations from experimental data. A significant improvement was achieved by introducing fit contact angles, which are substantially lower than the macroscopic experimental values and may be considered as approximate values of the microscopic contact angles. PMID- 12636691 TI - Faceting of a growing crystal surface by surface diffusion. AB - Consider faceting of a crystal surface caused by strongly anisotropic surface tension, driven by surface diffusion and accompanied by deposition (etching) due to fluxes normal to the surface. Nonlinear evolution equations describing the faceting of 1+1 and 2+1 crystal surfaces are studied analytically, by means of matched asymptotic expansions for small growth rates, and numerically otherwise. Stationary shapes and dynamics of faceted pyramidal structures are found as functions of the growth rate. In the 1+1 case it is shown that a solitary hill as well as periodic hill-and-valley solutions are unique, while solutions in the form of a solitary valley form a one-parameter family. It is found that with the increase of the growth rate, the faceting dynamics exhibits transitions from the power-law coarsening to the formation of pyramidal structures with a fixed average size and finally to spatiotemporally chaotic surfaces resembling the kinetic roughening. PMID- 12636692 TI - Nonequilibrium wetting transitions with short range forces. AB - We analyze within mean-field theory as well as numerically a Kardar-Parisi-Zhang equation that describes nonequilibrium wetting. Both complete and critical wettitng transitions were found and characterized in detail. For one-dimensional substrates the critical weting temperature is depressed by fluctuations. In addition, we have investigated a region in the space of parameters (temperature and chemical potential) where the wet and nonwet phases coexist. Finite-size scaling analysis of the interfacial detaching times indicates that the finite coexistence region survives in the thermodynamic limit. Within this region we have observed (stable or very long lived) structures related to spatiotemporal intermittency in other systems. In the interfacial representation these structures exhibit perfect triangular (pyramidal) patterns in one dimension (two dimensions), which are characterized by their slope and size distribution. PMID- 12636693 TI - Topological defects and nonhomogeneous melting of large two-dimensional Coulomb clusters. AB - The configurational and melting properties of large two-dimensional (2D) clusters of charged classical particles interacting with each other via the Coulomb potential are investigated through the Monte Carlo simulation technique. The particles are confined by a harmonic potential. For a large number of particles in the cluster (N>150), the configuration is determined by two competing effects, namely, the fact that in the center a hexagonal lattice is formed, which is the groundstate for an infinite 2D system, and the confinement that imposes its circular symmetry on the outer edge. As a result, a hexagonal Wigner lattice is formed in the central area while at the border of the cluster the particles are arranged in rings. In the transition region defects appear as dislocations and disclinations at the six corners of the hexagonal-shaped inner domain. Many different arrangements and types of defects are possible as metastable configurations with a slightly higher energy. The particle motion is found to be strongly related to the topological structure. Our results clearly show that the melting of the clusters starts near the geometry induced defects, and that three different melting temperatures can be defined corresponding to the melting of different regions in the cluster. PMID- 12636694 TI - Mechanism of sign inversion of spontaneous polarization in ferroelectric SmC* liquid crystals. AB - The ferroelectric liquid crystal FLC-117, which is known to exhibit at a certain temperature the sign inversion of spontaneous polarization P(s), is studied under an electric field. From the analysis of the electro-optic response and the direct texture observations, it is concluded that the inversion temperature of P(s) has the applied voltage dependence. To interpret our experimental result, we introduce a coupling term between the molecular dipole moments and the magnitude of the applied electric fields to the asymmetric rotational potential about the molecular long axis. The simulated results using this potential are in accordance with our experimental result. PMID- 12636695 TI - Experimental evidence for Sm-CG-->Sm-CP polymorphism in fluorinated bent-shaped mesogens. AB - We report the experimental results on pure fluorinated bent-shaped mesogens showing very unusual textures and electro-optical behavior. In the view of recent publications such behavior can be explained by triclinic symmetry of the mesophase (Sm-C(G)) formed by these compounds. Based on the results of the x-ray diffraction and electro-optical investigations, we give evidence for the Sm-C(G) ->Sm-CP polymorphism. PMID- 12636697 TI - Competition between the chiral smectic-C* and hexatic phases. AB - A theory of the phase transition from the smectic-C* to the hexatic phase based on the free energy with the two different Lifshitz terms is presented. Competition between the elastic energies of the tilt angle and of the hexatic order leads either to the single helicoidal structure, or to the double modulated solitonlike structure, and the transformation between both can occur. The bond order and the tilt angle suppress each other and at the transition to the low temperature hexatic phase a decrease of the tilt angle and an anomaly in the helical pitch occur. Approaching the hexatic phase transition the dielectric response is contributed mainly by the bond-order phason or (and) the tilt angle phason. While the bond-order phason frequency decreases the tilt angle frequency increases in the hexatic phase. Both situations are treated being observed in experiment. PMID- 12636696 TI - Effect of a quenched random field on a continuous symmetry breaking transition: nematic to smectic-A transition in octyloxycyanobiphenyl-aerosil dispersions. AB - High-resolution x-ray diffraction and ac-calorimetric experiments have been carried out on the liquid-crystal octyloxycyanobiphenyl in which aerosil particles are dispersed. The measurements were made over a temperature range around the bulk nematic to smectic-A transition temperature. At this transition the liquid crystal breaks translational symmetry in a single direction. The silica particles, which hydrogen bond together to form a very low density gel, provide the quenched disorder. The random gel leads to observable broadening of the x-ray reflection from the smectic layers. The structure factor is well described by modeling the effect of the aerosils as a quenched random field. Dispersed aerosils are thought to pin both the direction of the translational ordering and the position of the layers. The latter appears to have the greatest effect on the x-ray line shape. We show that the aerosil surface area, as verified by small-angle scattering, equates to the variance of the random field. Calorimetric results reveal substantial change in the specific heat peak associated with the nematic to smectic-A transition. As the concentration of aerosil increases, the specific heat peak remains sharp yet decreases in magnitude and shifts in temperature in a nonmonotonic fashion. In this regime, the critical exponent alpha becomes progressively smaller. For the samples with the largest concentrations of aerosil particles the C(p)(N-A) peak becomes highly smeared and shifts smoothly to lower temperatures. PMID- 12636698 TI - Order parameter measurements of dichroic dyes dissolved in smectic liquid crystals that tilt without layer contraction. AB - Measurements of the orientational order parameter of dissolved dichroic dyes are reported for two smectic-A liquid crystals that tilt in the presence of an in plane electric field without any decrease in the layer spacing. The dye order parameter is determined by measuring the anisotropic absorption of linearly polarized light. Different dyes are used and measurements are also performed on a smectic liquid crystal that tilts with the expected layer contraction to check how closely the measurements reflect the order parameter of the liquid crystal. The variation of the dye order parameter with electric field is in accordance with the recently proposed model of azimuthal disorder of the tilt angle direction, but the surprising finding is that the local dye orientational order parameter is significantly lower than for the smectic liquid crystal that tilts with the expected layer contraction. This suggests that another mechanism might be contributing significantly to the smectic order, one possibility of which is the tendency for different parts of these siloxane-containing molecules to segregate within each layer. Another possible explanation is that the azimuthal disorder is due to a modulated phase with a high enough density of defects to decrease the value of the local order parameter. PMID- 12636699 TI - Dynamics of electro-optical switching in the antiferroelectric B2 phase of an achiral bent-core shape compound. AB - A detailed study of the dynamics of electro-optical response has been carried out over the whole temperature range of the antiferroelectric B2 phase of a compound with bent-core shape molecules, a homolog (n=14) of the series 4-chloro-1,3 phenylene bis[4-(4-n-alkylphenylimino)benzoates]. Two types of stripe domains were observed with opposite handedness and simultaneous clock and anticlock motion of the director in the neighboring domains. The temperature dependence of the interlayer potential has been found from the threshold of the transition from the ground antiferroelectric (AF) state to the field-induced ferroelectric (F) state. The rotational viscosity gamma(phi) has been calculated from the dynamics of the field-induced azimuthal director switching between F-F and AF-F states and free relaxation of the director from F to AF state. The electro-optical response was also observed below the AF-F threshold. The latter was attributed to the soft mode distortion of the molecular tilt angle in the vicinity of the transition from the B2 phase to the isotropic phase. PMID- 12636700 TI - Dependence of film tension on the thickness of smectic films. AB - The film tension tau of free standing S(A) films has been measured for films with thicknesses between 2 and 150 layers. There is a clear increase of tau with the thickness for very thin films and a nonlinear slower increase for high thickness. The nonlinearity depends on the amount of liquid crystal accessible to the meniscus of the film during the drawing process. Several models are discussed that describe these effects. PMID- 12636701 TI - Interfaces of semi-infinite smectic liquid crystals and equations of state of infinite smectic stacks of semiflexible manifolds. AB - In this paper, we first elucidate the classical problem of the elastic free energy of a semi-infinite smectic-A liquid crystals, that fills the semispace above an interface (a boundary smectic layer) of a given shape. For the free energy of this interface, we obtain an effective interface Hamiltonian that takes into account the system discreteness introduced by the layered character of smectic-A liquid crystals. It is thus applicable to both short and long wavelength fluctuations of the interface shape. Next, we use our interface Hamiltonian to develop an efficient approach to the statistical mechanics of stacks of N semiflexible manifolds, such as two-dimensional smectic phases of long semiflexible polymers and three-dimensional lamellar fluid membrane phases. Within our approach, doing the practically interesting thermodynamic limit N--> infinity is reduced to considering a small stack, with just a few interacting manifolds, representing a subsystem of an infinite smectic. This dramatic reduction in the number of degrees of freedom is achieved by treating the first (the last) manifold of the small stack as an interface with the semi-infinite smectic medium below (above) the small stack. We illustrate our approach by considering in detail two-dimensional sterically stabilized smectic liquid crystals of long semiflexible polymers with hard-core repulsion. Smectic bulk (N= infinity ) equation of state and the universal constant characterizing entropic repulsion in these phases are obtained with a high accuracy from numerical simulations of small subsystems with just a few semiflexible polymers. PMID- 12636702 TI - Effect of confinement on phase-separation processes in a polymer blend observed by laser scanning confocal microscopy. AB - Structure self-assembling in the late stage spinodal decomposition of a polymer blend at its critical composition has been explored by laser-scanning confocal microscopy with particular emphasis on the effects of confinement (dimensionality) and preferential wetting of solid surface by one of the constituent polymers. A mixture of deuterated polybutadiene and polybutadiene (PB) with relatively narrow thickness (D congruent with 55 microm) was observed in three dimensions over the entire thickness. Formation of a wetting layer was clearly observed near the glass surface, while a bicontinuous structure evolved in the middle of the specimen. Global as well as local features of the phase separating structures were quantified by several structural parameters, e.g., characteristic length Lambda(m)(t), structure factor S(q), interfacial area per unit volume Sigma(t), probability densities of interfacial curvatures P(H,K;t), etc. (t is a phase-separation time). From the time evolution of these structural parameters, a deviation from the self-similar growth of a bicontinuous structure was found to occur at a transition time, t(tr), at which a scaled thickness, D/Lambda(m), approached unity. The breakdown of the self-similar growth was most sensitively observed by the local characteristics, i.e., Sigma(t) and P(H,K;t). On the other hand, the global characteristic, Lambda(m)(t), did not provide useful insight into the effects of dimensionality. It turned out that the bicontinuous structure, initially growing with dynamical self-similarity, eventually transformed into a "columnlike" structure (at t congruent with t(tr)) in which cylindrical PB-rich domains bridge the upper and lower PB wetting layers. PMID- 12636703 TI - Relaxation of a rubbed polystyrene surface. AB - The relaxation dynamics of a rubbed polystyrene (PS) surface have been characterized using infrared-visible sum frequency generation spectroscopy (SFG). The SFG results were compared with previous relaxation of retardation measurements, and the results show that the rubbed PS surface has the same T(g) as the bulk where T(g) is defined as tau(T(g))=5 s, however, the surface has a lower activation energy (deltaE) and a larger stretching exponent (beta(KWW)) than bulk PS. This indicates that the surface region relaxes more quickly than the bulk. The thickness of this region of lower deltaE and larger beta(KWW) is estimated to be roughly 12 nm. PMID- 12636704 TI - Probing polyelectrolyte elasticity using radial distribution function. AB - We study the effect of electrostatic interactions on the distribution function of the end-to-end distance of a single polyelectrolyte chain in the rodlike limit. The extent to which the radial distribution function of a polyelectrolyte is reproduced by that of a wormlike chain with an adjusted persistence length is investigated. Strong evidence is found for a universal scaling formula connecting the effective persistence length of a polyelectrolyte with its linear charge density and the Debye screening of its self-interaction. An alternative definition of the electrostatic persistence length is proposed based on matching of the maximum of the distribution with that of an effective wormlike chain, as opposed to the traditional matching of the first or the second moments of the distributions. It is shown that this definition provides a more accurate probe of the affinity of the distribution to that of the wormlike chains, as compared to the traditional definition. It is also found that the length of a polyelectrolyte segment can act as a crucial parameter in determining its elastic properties. PMID- 12636705 TI - Fast growth in phase-separating A-B-copolymer ternary mixtures with a chemical reaction. AB - We study the dynamics of phase separation of a binary A-B- polymer mixture with copolymer C, which is produced by the reaction of two counterpart reactive polymers A and B at the interface via the chemical reaction A+B right harpoon over left harpoon C. For low interfacial energy between the A and B phases, where the copolymer prefers to locate at interfaces, we show that the chemical reaction accelerates the phase separation of the system dramatically, because the backward reaction always drives the creation of immiscible A and B pairs at interfaces, which speed up the phase separation of the system, while the forward reaction process becomes more and more difficult as the interfaces are gradually saturated by copolymers. We also indicate that for a fixed chemical reaction rate constant, as the initial concentration of the copolymers increases, the domain growth at the late stage is speeded up as a result of the backward chemical reaction. However, when the interfacial energy is high, both forward and backward reactions coexist due to the occurrence of unsaturated interfaces, but the relative strength of reaction rates has no appreciable effect on domain growth during spinodal decomposition, because the interfacial energy dominates phase separation. PMID- 12636706 TI - Compact phases of polymers with hydrogen bonding. AB - We propose an off-lattice model for a self-avoiding homopolymer chain with two different competing attractive interactions, mimicking the hydrophobic effect and the hydrogen-bond formation, respectively. By means of Monte Carlo simulations, we are able to trace out the complete phase diagram for different values of the relative strengths of the two competing interactions. For strong enough hydrogen bonding, the ground state is a helical conformation, whereas with decreasing hydrogen-bonding strength, helices get eventually destabilized at low temperature in favor of more compact conformations resembling beta sheets appearing in the native structures of proteins. For weaker hydrogen bonding helices are not thermodynamically relevant anymore. PMID- 12636707 TI - Intramolecular caging in polybutadiene due to rotational barriers. AB - We present molecular dynamics simulations of a chemically realistic model of 1,4 polybutadiene and a freely rotating chain model derived from the first model by neglecting all dihedral potentials. We show that the presence of energy barriers hindering dihedral rotation leads to an intermediate plateau regime in the tagged particle mean-squared displacement reminiscent of the cage effect underlying the mode-coupling description of the liquid-glass transition. This intramolecular caging, however, occurs already at temperatures well above the glass transition regime. Because of its different physical origin, it also does not comply with the theoretical predictions of the mode-coupling theory. Consequences for the applicability of the mode-coupling theory to the glass transition in polymer melts are discussed. PMID- 12636708 TI - Dual-resolution coarse-grained simulation of the bisphenol-A-polycarbonate/nickel interface. AB - We present a dual-resolution coarse-graining scheme for efficient molecular dynamics simulations of bisphenol-A-polycarbonate (BP-A-PC) liquids in contact with a (111) nickel surface. The essential feature of this model is the strong adsorption of phenoxy chain ends, and the absence of adsorption by other parts of the chains. Details of how phenoxy chain ends interact with the nickel surface were extracted from Car-Parrinello molecular dynamics calculations of adsorption of phenol on nickel. These calculations show that phenol adsorption on nickel is short ranged (<3 A) and strongly dependent on the C1-C4 orientation of the ring. The structure of BP-A-PC prevents internal phenylene groups from interacting with the surface, due to steric hindrances from the noninteracting isopropylidenes. These dependencies are incorporated in the coarse-grained model of the BP-A-PC chain by resolving chain-terminating carbonate groups with atomistic detail, while the rest of the chain is represented by coarsened "beads." This allows specification of the C1-C4 orientation of the terminal phenoxy groups, while overall allowing for system equilibration with reasonable computer time. We simulate liquids of up to 240 chains of ten chemical repeat units, confined in a slit pore formed by two frozen (111) planes of atoms with the lattice spacing of nickel. We find that the strong adsorption of chain ends has a large effect on the liquid structure through a distance of more than two bulk radii of gyration from the surface. These effects are explained by a competition among single- and double-end adsorption, and dense packing. The structure of the interface less than 10 A from the wall is greatly sensitive to the orientational dependence of the phenoxy adsorption. PMID- 12636709 TI - Robustness and enhancement of neural synchronization by activity-dependent coupling. AB - We study the synchronization of two model neurons coupled through a synapse having an activity-dependent strength. Our synapse follows the rules of spike timing dependent plasticity. We show that this plasticity of the coupling between neurons produces enlarged frequency-locking zones and results in synchronization that is more rapid and much more robust against noise than classical synchronization arising from connections with constant strength. We also present a simple discrete map model that demonstrates the generality of the phenomenon. PMID- 12636710 TI - Nonlinear dependence of the delayed luminescence yield on the intensity of irradiation in the framework of a correlated soliton model. AB - We generalize the correlated soliton model in order to describe the delayed luminescence arising from biological systems after their exposition to the irradiation by relatively high dose (high intensity and/or long duration of irradiation). The quantum yield of the delayed luminescence is calculated as a function of the irradiation and is shown to depend nonlinearly on the intensity and dose of the irradiation. At relatively low intensity, the yield of luminescence increases with increasing dose, and monotonously reaches saturation. At high intensity of the irradiation, the yield of the photosystem under study is restricted from above by the concentration of photosystem units. As a result, the total yield of the delayed luminescence first increases with the dose till the maximum value that, in the general case, is less than the maximum number of available photosystem units. With further increase of the dose, the yield gradually decreases, reaching the saturation value at large dose of illumination. These results are obtained within the steady state approximation in the description of the luminescence kinetics. To check the applicability of this approximation at high levels and large time of illumination, the corrections to the steady state solution have been calculated, and shown to decrease exponentially with increase in time till the small finite constant value. The results of the theoretical model are shown to describe well the experimental data on the dose dependence of the quantum yield of the luminescence of algae Acetabularia acetabulum, for which the correlated soliton model describes well the kinetics of the delayed luminescence at low levels of irradiation. PMID- 12636711 TI - Modeling oscillatory microtubule polymerization. AB - Polymerization of microtubules is ubiquitous in biological cells and under certain conditions it becomes oscillatory in time. Here, simple reaction models are analyzed that capture such oscillations as well as the length distribution of microtubules. We assume reaction conditions that are stationary over many oscillation periods, and it is a Hopf bifurcation that leads to a persistent oscillatory microtubule polymerization in these models. Analytical expressions are derived for the threshold of the bifurcation and the oscillation frequency in terms of reaction rates, and typical trends of their parameter dependence are presented. Both, a catastrophe rate that depends on the density of guanosine triphosphate liganded tubulin dimers and a delay reaction, such as the depolymerization of shrinking microtubules or the decay of oligomers, support oscillations. For a tubulin dimer concentration below the threshold, oscillatory microtubule polymerization occurs transiently on the route to a stationary state, as shown by numerical solutions of the model equations. Close to threshold, a so called amplitude equation is derived and it is shown that the bifurcation to microtubule oscillations is supercritical. PMID- 12636712 TI - Energy resolution and dynamical heterogeneity effects on elastic incoherent neutron scattering from molecular systems. AB - Incoherent neutron scattering is widely used to probe picosecond-nanosecond time scale dynamics of molecular systems. In systems of spatially confined atoms the relatively high intensity of elastic incoherent neutron scattering is often used to obtain a first estimate of the dynamics present. For many complex systems, however, experimental elastic scattering is difficult to interpret unambiguously using analytical dynamical models that go beyond the determination of an average mean-square displacement. To circumvent this problem a description of the scattering is derived here that encompasses a variety of analytical models in a common framework. The framework describes the time-converged part of the dynamic structure factor [the elastic incoherent scattering function (EISF)] and lends itself to practical use by explicitly incorporating effects due to the finite energy resolution of the instrument used. The dependence of the elastic scattering on wave vector is examined, and it is shown how heterogeneity in the distribution of mean-square displacements can be related to deviations of the scattering from Gaussian behavior. In this case, a correction to fourth order in the scattering vector can be used to extract the variance of the distribution of mean-square displacements. The formalism is used in a discussion of measurements on dynamics accompanying the glass transition in molecular systems. By fitting to experimental data obtained on a protein solution the present methodology is used to show how the existence of a temperature-dependent relaxation frequency can lead to a transition in the measured mean-square displacement in the absence of an EISF change. PMID- 12636713 TI - Enhanced migration and ionic transport through membranes. AB - The effect of Coulomb forces upon transport enhancement of mobile ions, in the presence of slowly migrating charged polymeric chains, is investigated in a nonequilibrium regime brought about by a semipermeable membrane and a chemical reaction. By means of a numerical solution of the Nernst-Planck-Poisson equations, we predict the size of the effect, the conditions for positive or negative enhancement, and the dependence upon all relevant parameters. The limitations of the description of migration enhancement by an effective diffusion coefficient are also established. PMID- 12636714 TI - Solution of epidemic models with quenched transients. AB - We consider a model for single-season disease epidemics, with a delay (latent period) in the onset of infectivity and a decay ("quenching") in host susceptibility described by time-varying rates of primary and secondary infections. The classical susceptible-exposed-infected (SEI) model of epidemiology is a special case with constant rates. The decaying rates force the epidemics to slow down, and eventually stop in a "quenched transient" state that depends on the full history of the epidemic including its initial state. This equilibrium state is neutrally stable (i.e., has zero-value eigenvalues), and cannot be studied using standard equilibrium analysis. We introduce a method that gives an approximate analytical solution for the quenched state. The method uses an interpolation between two exactly solvable limits and applies to the whole, five-dimensional parameter space of the model. Some applications of the solutions for analysis of epidemics are given. PMID- 12636716 TI - Piecewise linear differential equations and integrate-and-fire neurons: insights from two-dimensional membrane models. AB - We derive and study two-dimensional generalizations of integrate-and-fire models which can be found from a piecewise linear idealization of the FitzHugh-Nagumo or Morris-Lecar model. These models give rise to new properties not present in one dimensional integrate-and-fire models. A detailed analytical study of the models is presented. In particular, (i) for the piecewise linear FitzHugh-Nagumo model, we determine analytically the bistability regime between stationary solutions and oscillations, that is, typical for class-II models. (ii) In the piecewise Morris Lecar model, we find a noncanonical class-I transition from a stationary state to oscillations with logarithmic dependence similar to that found for leaky integrate-and-fire models. (iii) Furthermore, we establish a relation to the recently proposed resonate-and-fire model and show that a short input current pulse can trigger several spikes. PMID- 12636715 TI - Role of photonic-crystal-type structures in the thermal regulation of a Lycaenid butterfly sister species pair. AB - One of the possible functions of the photonic-crystal structure found on the wing scales of some butterflies is investigated. The optical and electron microscopic investigation of two male butterflies-blue (colored) and brown (discolored) representing a sister species pair and originating from different altitudes, revealed that the blue color can be attributed unambiguously to the fine, spongelike medium, called "pepper-pot structure," present between the ridges and the cross ribs in the scales of the colored butterfly. Only traces of this structure can be found on the scales of the discolored butterfly. Other physical measurements, mainly optical reflectivity, transmission, and thermal measurements, are correlated with structural data and simulation results. The thermal measurements reveal that under identical illumination conditions the high altitude butterfly reaches a temperature 1.3-1.5 times the temperature reached by the low-altitude butterfly. This is attributed to the photonic-crystal-like behavior of the pepper-pot structure, which significantly reduces the penetration of light with wavelength in the blue region of the spectrum into the body of the scales. This sheds some light on the adaptation that enhances the survival chance of the butterfly in a cold environment rich in blue and UV radiation. PMID- 12636717 TI - Size exclusion and diffusion of fluoresceinated probes within collagen fibrils. AB - The diffusion of fluoresceinated probes inside single collagen fibrils was investigated by imaging the migration of fluorescence along the fibrils in oil and by monitoring fluorescence recovery after photobleaching (FRAP). Probes were excluded from the fibrils according to their size. Probes that were not excluded diffused in the fibrils, but FRAP occurred 6 x 10(-4) times more slowly than in water due to binding interactions between collagen and the probes. The dissociation constant of the fluorescein-collagen complex was determined (K(D)=1.8+/-0.1 microM). PMID- 12636718 TI - Theory of ac electrokinetic behavior of spheroidal cell suspensions with an intrinsic dispersion. AB - The dielectric dispersion, dielectrophoretic (DEP), and electrorotational (ER) spectra of spheroidal biological cell suspensions with an intrinsic dispersion in the constituent dielectric constants are investigated. By means of the spectral representation method, we express analytically the characteristic frequencies and dispersion strengths both for the effective dielectric constant and the Clausius Mossotti factor (CMF). We identify four and six characteristic frequencies for the effective dielectric spectra and CMF, respectively, all of them being dependent on the depolarization factor (or the cell shape). The analytical results allow us to examine the effects of the cell shape, the dispersion strength, and the intrinsic frequency on the dielectric dispersion, DEP, and ER spectra. Furthermore, we include the local-field effects due to the mutual interactions between cells in a dense suspension, and study the dependence of co field or antifield dispersion peaks on the volume fractions. PMID- 12636719 TI - Interstrand distance distribution of DNA near melting. AB - The distance distribution between complementary base pairs of the two strands of a DNA molecule is studied near the melting transition. Scaling arguments are presented for a generalized Poland-Scheraga-type model that includes self avoiding interactions. At the transition temperature and for a large distance r, the distribution decays as 1/r(kappa) with kappa=1+(c-2)/nu. Here nu is the self avoiding walk correlation length exponent and c is the exponent associated with the entropy of an open loop in the chain. Results for the distribution function just below the melting point are also presented. Numerical simulations that fully take into account the self-avoiding interactions are in good agreement with the scaling approach. PMID- 12636720 TI - Automated detection of a preseizure state based on a decrease in synchronization in intracranial electroencephalogram recordings from epilepsy patients. AB - The question whether information extracted from the electroencephalogram (EEG) of epilepsy patients can be used for the prediction of seizures has recently attracted much attention. Several studies have reported evidence for the existence of a preseizure state that can be detected using different measures derived from the theory of dynamical systems. Most of these studies, however, have neglected to sufficiently investigate the specificity of the observed effects or suffer from other methodological shortcomings. In this paper we present an automated technique for the detection of a preseizure state from EEG recordings using two different measures for synchronization between recording sites, namely, the mean phase coherence as a measure for phase synchronization and the maximum linear cross correlation as a measure for lag synchronization. Based on the observation of characteristic drops in synchronization prior to seizure onset, we used this phenomenon for the characterization of a preseizure state and its distinction from the remaining seizure-free interval. After optimizing our technique on a group of 10 patients with temporal lobe epilepsy we obtained a successful detection of a preseizure state prior to 12 out of 14 analyzed seizures for both measures at a very high specificity as tested on recordings from the seizure-free interval. After checking for in-sample overtraining via cross validation, we applied a surrogate test to validate the observed predictability. Based on our results, we discuss the differences of the two synchronization measures in terms of the dynamics underlying seizure generation in focal epilepsies. PMID- 12636721 TI - Evolution equation of population genetics: relation to the density-matrix theory of quasiparticles with general dispersion laws. AB - The Waxman-Peck theory of population genetics is discussed in regard of soil bacteria. Each bacterium is understood as a carrier of a phenotypic parameter p. The central objective is the calculation of the probability density with respect to p, Phi(p,t;p(0)), of the carriers living at time t>0, provided that initially at t(0)=0, all bacteria carried the phenotypic parameter p(0)=0. The theory involves two small parameters: the mutation probability mu and a parameter gamma involved in a function w(p) defining the fitness of the bacteria to survive the generation time tau and give birth to an offspring. The mutation from a state p to a state q is defined by a Gaussian with a dispersion sigma(2)(m). The author focuses our attention on a function phi(p,t) which determines uniquely the function Phi(p,t;p(0)) and satisfies a linear equation (Waxman's equation). The Green function of this equation is mathematically identical with the one-particle Bloch density matrix, where mu characterizes the order of magnitude of the potential energy. (In the x representation, the potential energy is proportional to the inverted Gaussian with the dispersion sigma(2)(m)). The author solves Waxman's equation in the standard style of a perturbation theory and discusses how the solution depends on the choice of the fitness function w(p). In a sense, the function c(p)=1-w(p)/w(0) is analogous to the dispersion function E(p) of fictitious quasiparticles. In contrast to Waxman's approximation, where c(p) was taken as a quadratic function, c(p) approximately gammap(2), the author exemplifies the problem with another function, c(p)=gamma[1-exp(-ap(2))], where gamma is small but a may be large. The author shows that the use of this function in the theory of the population genetics is the same as the use of a nonparabolic dispersion law E=E(p) in the density-matrix theory. With a general function c(p), the distribution function Phi(p,t;0) is composed of a delta-function component, N(t)delta(p), and a blurred component. When discussing the limiting transition for t--> infinity, the author shows that his function c(p) implies that N(t)-->N( infinity ) not equal 0 in contrast with the asymptotics N(t)-->0 resulting from the use of Waxman's function c(p) approximately p(2). PMID- 12636722 TI - Statistical physics of RNA folding. AB - We discuss the physics of RNA as described by its secondary structure. We examine the static properties of a homogeneous RNA model that includes pairing and base stacking energies as well as entropic costs for internal loops. For large enough loop costs the model exhibits a thermal denaturation transition which we analyze in terms of the radius of gyration. We point out an inconsistency in the standard approach to RNA secondary structure prediction for large molecules. Under an external force a second-order phase transition between a globular and an extended phase takes place. A Harris-type criterion shows that sequence disorder does not affect the correlation length exponent while the other critical exponents are modified in the glass phase. However, at high temperatures, on a coarse-grained level, disordered RNA is well described by a homogeneous model. The characteristics of force-extension curves are discussed as a function of the energy parameters. We show that the force transition is always second order. A reentrance phenomenon relevant for real disordered RNA is predicted. PMID- 12636723 TI - Modeling postshock evolution of large electropores. AB - The Smoluchowski equation (SE), which describes the evolution of pores created by electric shocks, cannot be applied to modeling large and long-lived pores for two reasons: (1) it does not predict pores of radius above 20 nm without also predicting membrane rupture; (2) it does not predict postshock growth of pores. This study proposes a model in which pores are coupled by membrane tension, resulting in a nonlinear generalization of SE. The predictions of the model are explored using examples of homogeneous (all pore radii r are equal) and heterogeneous (00 for times less than the Ehrenfest time tau(E)=1/2lambda0(-1)|ln Planck|. It is only after the Ehrenfest time that the average decay is representative for a typical initial condition. PMID- 12636736 TI - Structural transitions in one-dimensionally confined one-component plasmas. AB - Layered structures occurring in strongly coupled one-dimensionally confined one component plasmas are studied by means of Monte Carlo simulations. A series of structural transitions with alternating square and hexagonal symmetry is observed with increasing the width of the system. For closer interlayer separation, the square type of lattice (bcc in [100] or [110] plane) is dominating, whereas for wider separations the hexagonal (fcc in [111] plane or hcp) patterns are observed. PMID- 12636737 TI - Dark and gray strong dispersion-managed solitons. AB - Dark and gray solitons in communication systems with strong dispersion management (DM) are obtained. These new modes are characterized by a decaying oscillatory background. Unlike the bright DM solitons in which the oscillations are observed only on a logarithmic scale, here the oscillations are dominant on a linear scale and become very strong for moderate map strength. PMID- 12636738 TI - Electromagnetic waves focused by a negative-index planar lens. AB - We demonstrate that negative refraction occurs for both cw and pulsed electromagnetic waves when traversing from a "right-handed" (index > 0) to a "left-handed" (index < 0) material (LHM) which has causal dispersive intrinsic properties. We also demonstrate that a divergent line source spaced a distance H in front of a planar LHM slab and excited by either an impulse cw or a Gaussian frequency pulse is imaged at a distance H away, inside the LHM, and at H to the other side of the slab. The image size is approximately lambda consistent with limitations dictated by wave optics. We find no evidence of evanescent mode amplification. The studies were performed using numerical experiments with finite difference time domain solutions and incorporating a causal Lorentzian form for the frequency-dependent material properties. PMID- 12636739 TI - Modulational instability arising from collective Rayleigh scattering. AB - It is shown that under certain conditions a collection of dielectric Rayleigh particles suspended in a viscous medium and enclosed in a bidirectional ring cavity pumped by a strong laser field can produce a new modulational instability transverse to the wave-propagation direction. The source of the instability is collective Rayleigh scattering i.e., the spontaneous formation of periodic longitudinal particle-density modulations and a backscattered optical field. Using a linear stability analysis a dispersion relation is derived which determines the region of parameter space in which modulational instability of the backscattered field and the particle distribution occurs. In the linear regime the pump is modulationally stable. A numerical analysis is carried out to observe the dynamics of the interaction in the nonlinear regime. In the nonlinear regime the pump field also becomes modulationally unstable and strong pump depletion occurs. PMID- 12636740 TI - Hilbert phase analysis of the dynamics of a semiconductor laser with optical feedback. AB - The nonlinear dynamics of a semiconductor laser with an optical feedback is studied using Hilbert phase analysis, which reveals interesting facets of the nonlinear dynamical behavior, including the formation and interaction of external cavity modes with increasing external feedback. Here we report measurements on two illustrative cases with very different dynamics; the laser is first biased just near threshold, and then far above threshold. We observe 2pi phase jumps at intervals that are multiples of the external cavity round-trip time, indicating the interaction and transfer of energy between external cavity modes. PMID- 12636741 TI - Lattice Boltzmann model for photonic band gap materials. AB - An efficient technique for computing photonic band structure and defect modes is proposed based on the lattice Boltzmann model. Physically, it is a scheme based on the kinetics of the virtual microscopic process, rather than a solution of the macroscopic Maxwell equations. The method has significant advantages of being naturally suited for massively parallel machine, as well as speed and convenience, providing another methodology for photonic band gap materials and, also, for general electromagnetic scattering problems in open region when incorporated with the perfectly matched layer technique. PMID- 12636742 TI - Rejection-free Monte Carlo algorithms for models with continuous degrees of freedom. AB - We construct a rejection-free Monte Carlo algorithm for a system with continuous degrees of freedom. We illustrate the algorithm by applying it to the classical three-dimensional Heisenberg model with canonical Metropolis dynamics. We obtain the lifetime of the metastable state following a reversal of the external magnetic field. Our rejection-free algorithm obtains results in agreement with a direct implementation of the Metropolis dynamic and requires orders of magnitude less computational time at low temperatures. The treatment is general and can be extended to other dynamics and other systems with continuous degrees of freedom. PMID- 12636743 TI - Competing tunneling trajectories in a two-dimensional potential with variable topology as a model for quantum bifurcations. AB - We present a path-integral approach to treat a two-dimensional model of a quantum bifurcation. The model potential has two equivalent minima separated by one or two saddle points, depending on the value of a continuous parameter. Tunneling is, therefore, realized either along one trajectory or along two equivalent paths. The zero-point fluctuations smear out the sharp transition between these two regimes and lead to a certain crossover behavior. When the two saddle points are inequivalent one can also have a first order transition related to the fact that one of the two trajectories becomes unstable. We illustrate these results by numerical investigations. Even though a specific model is investigated here, the approach is quite general and has potential applicability for various systems in physics and chemistry exhibiting multistability and tunneling phenomena. PMID- 12636744 TI - Fractal dimension of 3-blocks in four-, five-, and six-dimensional percolation systems. AB - Using Monte Carlo simulations, we study the distributions of the 3-block mass N3 in four-, five-, and six-dimensional percolation systems. Because the probability of creating large 3-blocks in these dimensions is very small, we use a "go with the winners" method of statistical enhancement to simulate configurations having probability as small as 10(-30). In earlier work, the fractal dimensions of 3 blocks, d(3), in 2D (two dimensional) and 3D were found to be 1.20+/-0.1 and 1.15+/-0.1, respectively, consistent with the possibility that the fractal dimension might be the same in all dimensions. We find that the fractal dimension of 3-blocks decreases rapidly in higher dimensions, and estimate d(3)=0.7+/-0.2 (4D) and 0.5+/-0.2 (5D). At the upper critical dimension of percolation, d(c)=6, our simulations are consistent with d(3)=0 with logarithmic corrections to power law scaling. PMID- 12636745 TI - Small-world effects in the majority-vote model. AB - We investigate the majority-vote model on small-world networks by rewiring the two-dimensional square lattice. We observe that the introduction of long-range interactions does not remove the critical character of the model, that is, the system still exhibits a well-defined phase transition. However, we find that now the critical point is a monotonically increasing function of the rewiring probability. Moreover, we find that small-world effects change the class of universality of the model. PMID- 12636746 TI - Exotic trees. AB - We discuss the scaling properties of free branched polymers. The scaling behavior of the model is classified by the Hausdorff dimensions for the internal geometry, d(L) and d(H), and for the external one, D(L) and D(H). The dimensions d(H) and D(H) characterize the behavior for long distances, while d(L) and D(L) for short distances. We show that the internal Hausdorff dimension is d(L)=2 for generic and scale-free trees, contrary to d(H), which is known to be equal to 2 for generic trees and to vary between 2 and infinity for scale-free trees. We show that the external Hausdorff dimension D(H) is directly related to the internal one as D(H)=alphad(H), where alpha is the stability index of the embedding weights for the nearest-vertex interactions. The index is alpha=2 for weights from the Gaussian domain of attraction and 01 (x>>1). We then show within the independent interval approximation (IIA) that the distribution n(k,t) of separation k between two consecutive persistent spins at time t has the asymptotic scaling form n(k,t)=t( 2phi)g(t,k/t(phi)), where the dynamical exponent has the form phi=max(1/2,theta). The behavior of the scaling function for large and small values of the arguments is found analytically. We find that for small separations k<>t(phi), g(t,x) decays exponentially with x. The unusual dynamical scaling form and the behavior of the scaling function is supported by numerical simulations. PMID- 12636757 TI - Metastable configurations on the Bethe lattice. AB - We present a general analytic method to compute the number of metastable configurations as a function of the energy for a system of interacting Ising spins on the Bethe lattice. Our approach is based on the cavity method. We apply it to the case of ferromagnetic interactions, and also to the binary and Gaussian spin glasses. Most of our results are obtained within the replica symmetric ansatz, but we illustrate how replica symmetry breaking can be performed. PMID- 12636758 TI - Interface dynamics and solute trapping in alloy solidification with density change. AB - We present a phase-field model for the solidification of a binary alloy, which incorporates hydrodynamic effects due to the different densities of the solid and liquid phases. We start from a generalized thermodynamic potential with squared gradient terms for the associated fields; the condition of local positive entropy production is then utilized to derive a set of equations that drive the system towards equilibrium. The model has been numerically solved in one dimension, to investigate the effects of the flow field on the interface dynamics. We observed that solute trapping is almost unaffected by the fluid advection, while the interface mobility is strongly reduced as the fluid velocity increases. This reflects on the dependence of the interface temperature T(I) on the growth rate v(I): the region of the unstable (ascending) branch is reduced, and the maximum of the T(I)(v(I)) curve is shifted towards lower velocities. PMID- 12636759 TI - Star-graph expansions for bond-diluted Potts models. AB - We derive high-temperature series expansions for the free energy and the susceptibility of random-bond q-state Potts models on hypercubic lattices using a star-graph expansion technique. This method enables the exact calculation of quenched disorder averages for arbitrary uncorrelated coupling distributions. Moreover, we can keep the disorder strength p as well as the dimension d as symbolic parameters. By applying several series analysis techniques to the new series expansions, one can scan large regions of the (p,d) parameter space for any value of q. For the bond-diluted four-state Potts model in three dimensions, which exhibits a rather strong first-order phase transition in the undiluted case, we present results for the transition temperature and the effective critical exponent gamma as a function of p as obtained from the analysis of susceptibility series up to order 18. A comparison with recent Monte Carlo data [Chatelain et al., Phys. Rev. E 64, 036120 (2001)] shows signals for the softening to a second-order transition at finite disorder strength. PMID- 12636760 TI - Magnetization reversal times in the two-dimensional Ising model. AB - We present a theoretical framework which is generally applicable to the study of time scales of activated processes in systems with Brownian type dynamics. This framework is applied to a prototype system: magnetization reversal times in the two-dimensional Ising model. Direct simulation results for the magnetization reversal times, spanning more than five orders of magnitude, are compared with theoretical predictions; the two agree in most cases within 20%. PMID- 12636761 TI - Nonequilibrium transitions in complex networks: a model of social interaction. AB - We analyze the nonequilibrium order-disorder transition of Axelrod's model of social interaction in several complex networks. In a small-world network, we find a transition between an ordered homogeneous state and a disordered state. The transition point is shifted by the degree of spatial disorder of the underlying network, the network disorder favoring ordered configurations. In random scale free networks the transition is only observed for finite size systems, showing system size scaling, while in the thermodynamic limit only ordered configurations are always obtained. Thus, in the thermodynamic limit the transition disappears. However, in structured scale-free networks, the phase transition between an ordered and a disordered phase is restored. PMID- 12636762 TI - Measuring nonequilibrium temperature of forced oscillators. AB - The meaning of temperature in nonequilibrium thermodynamics is considered by using a forced harmonic oscillator in a heat bath, where we have two effective temperatures for the position and the momentum, respectively. We propose a concrete model of a thermometer to testify the validity of these different temperatures from the operational point of view. It is found that the measured temperature depends on a specific form of interaction between the system and a thermometer, which means that the zeroth law of thermodynamics cannot be immediately extended to nonequilibrium cases. PMID- 12636763 TI - Phase behavior of confined symmetric binary mixtures. AB - We employ mean-field lattice density functional theory to investigate the phase behavior of a binary (A-B) mixture confined to nanoscopic slit pores with chemically homogeneous walls. We consider only nearest-neighbor interactions in symmetric mixtures, where epsilon(AA)=epsilon(BB) not equal epsilon(AB) and epsilon is a measure of attraction between molecules of like (subscripts AA and BB) and unlike species (subscript AB), respectively. In addition, molecules are exposed to short-range attraction by the substrates separated by z lattice planes where epsilon(W) is the relevant coupling parameter. Moreover, the chemical potentials of both components are the same, that is, mu(A)=mu(B)=mu. In thermodynamic equilibrium (for fixed temperature T and chemical potential mu) the grand-potential density omega[rho,m] (rho identical with [rho(1),...,rho(z)], m identical with [m(1),...,m(z)]) assumes a global minimum which we find by minimizing omega numerically with respect to the order parameters rho(l) identical with rho(A)(l)+rho(B)(l) (total local density) and m(l) identical with (rho(A)(l)-rho(B)(l))/rho(l) (local "miscibility") at lattice plane l parallel to the pore walls. By varying epsilon(AB) three generic types of bulk phase diagrams are observed. On account of confinement (i.e., by varying epsilon(W) as well as z) one may switch between these different types of phase diagrams. This may have profound practical repercussions for experimental nanophase separation since depending on pore width and chemical nature of its walls a bulk gas mixture may undergo capillary condensation and form either a stable mixed or demixed liquid phase. PMID- 12636764 TI - Critical phenomena in networks. AB - We develop a phenomenological theory of critical phenomena in networks with an arbitrary distribution of connections P(k). The theory shows that the critical behavior depends in a crucial way on the form of P(k) and differs strongly from the standard mean-field behavior. The critical behavior observed in various networks is analyzed and found to be in agreement with theory. PMID- 12636765 TI - Random series and discrete path integral methods: The Levy-Ciesielski implementation. AB - We perform a thorough analysis of the relationship between discrete and series representation path integral methods, which are the main numerical techniques used in connection with the Feynman-Kac formula. First, an interpretation of the so-called standard discrete path integral methods is derived by direct discretization of the Feynman-Kac formula. Second, we consider a particular random series technique based upon the Levy-Ciesielski representation of the Brownian bridge and analyze its main implementations, namely the primitive, the partial averaging, and the reweighted versions. It is shown that the n=2(k)-1 subsequence of each of these methods can also be interpreted as a discrete path integral method with appropriate short-time approximations. We therefore establish a direct connection between the discrete and the random series approaches. In the end, we give sharp estimates on the rates of convergence of the partial averaging and the reweighted Levy-Ciesielski random series approach for sufficiently smooth potentials. The asymptotic rates of convergence are found to be O(1/n(2)), in agreement with the rates of convergence of the best standard discrete path integral techniques. PMID- 12636766 TI - Introducing small-world network effects to critical dynamics. AB - We analytically investigate the kinetic Gaussian model and the one-dimensional kinetic Ising model of two typical small-world networks (SWN), the adding type and the rewiring type. The general approaches and some basic equations are systematically formulated. The rigorous investigation of the Glauber-type kinetic Gaussian model shows the mean-field-like global influence on the dynamic evolution of the individual spins. Accordingly a simplified method is presented and tested, which is believed to be a good choice for the mean-field transition widely (in fact, without exception so far) observed for SWN. It yields the evolving equation of the Kawasaki-type Gaussian model. In the one-dimensional Ising model, the p dependence of the critical point is analytically obtained and the nonexistence of such a threshold p(c), for a finite-temperature transition, is confirmed. The static critical exponents gamma and beta are in accordance with the results of the recent Monte Carlo simulations, and also with the mean-field critical behavior of the system. We also prove that the SWN effect does not change the dynamic critical exponent z=2 for this model. The observed influence of the long-range randomness on the critical point indicates two obviously different hidden mechanisms. PMID- 12636767 TI - Mixing patterns in networks. AB - We study assortative mixing in networks, the tendency for vertices in networks to be connected to other vertices that are like (or unlike) them in some way. We consider mixing according to discrete characteristics such as language or race in social networks and scalar characteristics such as age. As a special example of the latter we consider mixing according to vertex degree, i.e., according to the number of connections vertices have to other vertices: do gregarious people tend to associate with other gregarious people? We propose a number of measures of assortative mixing appropriate to the various mixing types, and apply them to a variety of real-world networks, showing that assortative mixing is a pervasive phenomenon found in many networks. We also propose several models of assortatively mixed networks, both analytic ones based on generating function methods, and numerical ones based on Monte Carlo graph generation techniques. We use these models to probe the properties of networks as their level of assortativity is varied. In the particular case of mixing by degree, we find strong variation with assortativity in the connectivity of the network and in the resilience of the network to the removal of vertices. PMID- 12636768 TI - Classical nucleation theory revisited. AB - A field-theoretic derivation of the correction to classical nucleation theory due to translational invariance of a nucleating droplet is proposed. The correction is derived from a functional integral representation of the classical partition function, where the two-body interaction potential is decomposed into a short range repulsive part and a long-range attractive part. The functional integral is evaluated in the mean-field approximation, and the spatially nonuniform density solution of the Euler-Lagrange equation is approximated by a physically motivated hyperbolic tangent profile. Leading-order effects of the nonlocal attractive interaction are highlighted through a density-gradient expansion. The capillarity approximation to the droplet free energy of formation is obtained by performing a density resummation of the uniform state, low-density expansion of the Helmholtz free energy density, and by retaining the leading-order density-gradient term. The resulting translational-invariance correction modifies the droplet free energy by an additive mixing-entropy term. The additional contribution, which contains a logarithmic correction to the surface-energy term, defines a scaling volume that depends on the range of the coarse-grained attractive potential. PMID- 12636769 TI - Subdiffusion and localization in the one-dimensional trap model. AB - We study a one-dimensional generalization of the exponential trap model using both numerical simulations and analytical approximations. We obtain the asymptotic shape of the average diffusion front in the subdiffusive phase. Our central result concerns the localization properties. We find the dynamical participation ratios to be finite, but different from their equilibrium counterparts. Therefore, the idea of a partial equilibrium within the limited region of space explored by the walk is not exact, even for long times where each site is visited a very large number of times. We discuss the physical origin of this discrepancy, and characterize the full distribution of dynamical weights. We also study two different two-time correlation functions, which exhibit different aging properties: one is "sub aging" whereas the other one shows "full aging," therefore, two diverging time scales appear in this model. We give intuitive arguments and simple analytical approximations that account for these differences, and obtain new predictions for the asymptotic (short-time and long time) behavior of the scaling functions. Finally, we discuss the issue of multiple time scalings in this model. PMID- 12636770 TI - Controlling chaos to solutions with complex eigenvalues. AB - We derive formulas for parameter and variable perturbations to control chaos using linearized dynamics. They are available irrespective of the dimension of the system, the number of perturbed parameters or variables, and the kinds of eigenvalues of the linearized dynamics. We illustrate this using the two coupled Duffing oscillators and the two coupled standard maps. PMID- 12636771 TI - Synchronization and desynchronization under the influence of quasiperiodic forcing. AB - We study the influence of quasiperiodic forcing on synchronization and desynchronization using two coupled quasiperiodically forced logistic maps as a paradigm. We show that due to the forcing the synchronization region in parameter space shrinks. The loss of transverse stability of the synchronized attractors leads to desynchronization. Two types of such blowout bifurcations are described, namely, the blowout bifurcations of synchronized quasiperiodic motion on invariant curves and synchronized strange nonchaotic attractors, both yielding desynchronized chaotic attractors. PMID- 12636772 TI - Statistics and characteristics of spatiotemporally rare intense events in complex Ginzburg-Landau models. AB - We study the statistics and characteristics of rare intense events in two types of two-dimensional complex Ginzburg-Landau (CGL) equation based models. Our numerical simulations show finite amplitude collapselike solutions which approach the infinite amplitude solutions of the nonlinear Schrodinger equation in an appropriate parameter regime. We also determine the probability distribution function of the amplitude of the CGL solutions, which is found to have enhanced (as compared to Gaussian) probability for the amplitude to be large. Our results suggest a general picture in which an incoherent background of weakly interacting waves, occasionally, "by chance," initiates intense, coherent, self-reinforcing, highly nonlinear events. PMID- 12636774 TI - Estimating topological entropy via a symbolic data compression technique. AB - We estimate topological entropy via symbolic dynamics using a data compression technique called the context-tree weighting method. Unlike other symbolic dynamical approaches, which often have to choose ad hoc parameters such as the depth of a tree, the context-tree weighting method is almost parameter-free and infers the transition structure of the system as well as transition probabilities. Our examples, including a Markov model, the logistic map, and the Henon map, demonstrate that the convergence is fast: one obtains the theoretically correct topological entropy with a relatively short symbolic sequence. PMID- 12636773 TI - Exceptional points and double poles of the S matrix. AB - Exceptional points and double poles of the S matrix are both characterized by the coalescence of a pair of eigenvalues. In the first case, the coalescence causes a defect of the Hilbert space. In the second case, this is not so as shown in previous papers. Mathematically, the reason for this difference is the biorthogonality of the eigenfunctions of a non-Hermitian operator that is ignored in the first case. The consequences for the topological structure of the Hilbert space are studied and compared with existing experimental data. PMID- 12636775 TI - Stadium billiard with moving walls. AB - We study the evolution of the energy distribution for a stadium with moving walls. We consider a one period driving cycle, which is characterized by an amplitude A and a wall velocity V. This evolving energy distribution has both "parametric" and "stochastic" components. The latter are important for the theory of quantum irreversibility and dissipation in driven mesoscopic devices. For an extremely slow wall velocity V, the spreading mechanism is dominated by transitions between neighboring levels, while for larger (nonadiabatic) velocities, the spreading mechanism has both perturbative and nonperturbative features. We present a numerical study which is aimed at identifying the latter features. A procedure is developed for the determination of the various V regimes. The possible implications of linear response theory are discussed. PMID- 12636776 TI - Chaotic communications that are difficult to detect. AB - Work on self-synchronizing systems for communications has had limited practicality because the chaotic signals were not as easy to detect in the presence of noise as conventional spread-spectrum signals. This difficulty may actually be an advantage in some cases, where one wants to conceal the existence of the communications signal. Conventional communications signals are cyclostationary; while they may look random, they have statistical properties that vary periodically. One may design chaotic communication signals that lack this cyclostationary property, and therefore are harder to detect. PMID- 12636777 TI - Measuring billiard eigenfunctions with arbitrary trajectories. AB - We propose a method of measuring approximate quantum eigenfunctions in polygonalized billiard geometries, based on a quasiclassical evolution operator having a (smoothened) Perron-Frobenius kernel modulated by a phase arising from quantum considerations. Using a plane wave ansatz, we show that the condition under which this is an eigenfunction of the quasiclassical operator is identical to the condition for it to be an eigenfunction of the Schrodinger equation for polygonalized billiards. Finally, we demonstrate this technique by determining the quasiclassical eigenfunctions of the polygonalized stadium billiard using arbitrary trajectories and comparing this with the exact quantum stadium eigenfunctions. PMID- 12636778 TI - General stability analysis of synchronized dynamics in coupled systems. AB - We consider the stability of synchronized states (including equilibrium point, periodic orbit, or chaotic attractor) in arbitrarily coupled dynamical systems (maps or ordinary differential equations). We develop a general approach, based on the master stability function and Gershgorin disk theory, to yield constraints on the coupling strengths to ensure the stability of synchronized dynamics. Systems with specific coupling schemes are used as examples to illustrate our general method. PMID- 12636779 TI - Noise-induced unstable dimension variability and transition to chaos in random dynamical systems. AB - Results are reported concerning the transition to chaos in random dynamical systems. In particular, situations are considered where a periodic attractor coexists with a nonattracting chaotic saddle, which can be expected in any periodic window of a nonlinear dynamical system. Under noise, the asymptotic attractor of the system can become chaotic, as characterized by the appearance of a positive Lyapunov exponent. Generic features of the transition include the following: (1) the noisy chaotic attractor is necessarily nonhyperbolic as there are periodic orbits embedded in it with distinct numbers of unstable directions (unstable dimension variability), and this nonhyperbolicity develops as soon as the attractor becomes chaotic; (2) for systems described by differential equations, the unstable dimension variability destroys the neutral direction of the flow in the sense that there is no longer a zero Lyapunov exponent after the noisy attractor becomes chaotic; and (3) the largest Lyapunov exponent becomes positive from zero in a continuous manner, and its scaling with the variation of the noise amplitude is algebraic. Formulas for the scaling exponent are derived in all dimensions. Numerical support using both low- and high-dimensional systems is provided. PMID- 12636780 TI - Density of states for vibrations of fractal drums. AB - Vibrations of membranes with fractal boundaries (fractal drums) are investigated. Numerical results are presented for Koch drums of fractal dimension D(f)=3/2 at prefractal generations 1-3, and for Koch snowflake drums (D(f)=ln 4/ln 3) at generations 3 and 4. The results show that the low-frequency integrated densities of states (IDOS's) of the drums are well approximated by a two-term asymptotic of the form given by the modified Weyl-Berry (MWB) conjecture, which predicts a correction of DeltaN(Omega) proportional, variant Omega(D(f)) to the leading order Weyl term. In the high-frequency regime, where the half wavelength is smaller than the smallest features of the prefractal perimeter, the two-term Weyl asymptotic is applicable, with DeltaN(Omega) approximately Omega. The results also indicate that oscillations in DeltaN(Omega) arise due to localization of the wave amplitude near the prefractal perimeter. It is argued that for a self similar fractal boundary, the amplitude of the oscillations is asymptotically proportional to Omega(D(f)), which implies an O(Omega(D(f))), rather than the conjectured o(Omega(D(f))), error term for the asymptotic IDOS given by the MWB conjecture. PMID- 12636781 TI - Chaotic-time-series reconstruction by the Bayesian paradigm: right results by wrong methods. AB - Recently, papers have appeared that champion the Bayesian approach to the analysis of experimental data. From reading these papers, the physicist could be forgiven for believing that Bayesian methods reveal deep truths about physical systems and are the correct paradigm for the analysis of all experimental data. This paper makes a contrary argument and is deliberately provocative. It is argued that the Bayesian approach to reconstruction of chaotic time series is fundamentally flawed, and the apparent successes result not from any degree of correctness of the paradigm, but by an accidental and unintended property of an algorithm. We also argue that (non-Bayesian) shadowing techniques provide all the information the erroneous Bayesian methods obtain, but much more efficiently, and also provide a wealth of additional useful information. PMID- 12636782 TI - Chaotic sound waves in a regular billiard. AB - We present experimental results for the ultrasound transmission spectra and standing wave patterns of a rectangular block of fused quartz. A comparison is made between our data and an approximation of the theoretical staircase function for three-dimensional isotropic elasticity. The main emphasis of our study is on the role of mode conversion in regular ray-splitting billiards. We present the fluctuation statistics and find that these are described by the Gaussian orthogonal ensemble of random matrix theory, despite the fact that the system is not classically chaotic, as demonstrated with numerical simulation. Using temperature perturbation, we find that the vast majority of the resonances are mixtures of transverse and longitudinal wave motion, yet a small number of special resonances remain pure. We further illuminate this by presenting standing wave patterns measured on one face of the block. PMID- 12636783 TI - Optimization and stability boundaries for the synchronization of semiconductor lasers with external optical feedback. AB - We perform a stability and optimization analysis for the synchronization of unidirectionally coupled external-cavity semiconductor lasers. Using rigorous stability criteria, we qualitatively derive the boundaries of the high-quality synchronization basin. The underlying influence of Hopf bifurcations on the stability of the synchronization manifold is also investigated. PMID- 12636784 TI - Is the concept of the non-Hermitian effective Hamiltonian relevant in the case of potential scattering? AB - We examine the notion and properties of the non-Hermitian effective Hamiltonian of an unstable system using as an example potential resonance scattering with a fixed angular momentum. We present a consistent self-adjoint formulation of the problem of scattering on a finite-range potential, which is based on the separation of the configuration space into two segments, internal and external. The scattering amplitude is expressed in terms of the resolvent of a non Hermitian operator H. The explicit form of this operator depends on both the radius of separation and the boundary conditions at this place, which can be chosen in many different ways. We discuss this freedom and show explicitly that the physical scattering amplitude is, nevertheless, unique, although not all choices are equally adequate from the physical point of view. The energy dependent operator H should not be confused with the non-Hermitian effective Hamiltonian H(eff) which is usually exploited to describe interference of overlapping resonances. We note that the simple Breit-Wigner approximation is as a rule valid for any individual resonance in the case of few-channel scattering on a flat billiardlike cavity, leaving no room for nontrivial H(eff) to appear. The physics is appreciably richer in the case of an open chain of L connected similar cavities whose spectrum has a band structure. For a fixed band of L overlapping resonances, the smooth energy dependence of H can be ignored so that the constant LxL submatrix H(eff) approximately describes the time evolution of the chain in the energy domain of the band and the complex eigenvalues of H(eff) define the energies and widths of the resonances. We apply the developed formalism to the problem of a chain of L delta barriers, whose solution is also found independently in a closed form. We construct H(eff) for the two commonly considered types of boundary conditions (Neumann and Dirichlet) for the internal motion. Although the final results are in perfect coincidence, somewhat different physical patterns arise of the trend of the system with growing openness. Formation in the outer well of a short-lived doorway state shifted in energy is explicitly demonstrated together with the appearance of L-1 long-lived states trapped in the inner part of the chain. PMID- 12636785 TI - Synchronization of coupled rotators: Josephson junction ladders and the locally coupled Kuramoto model. AB - We show that the resistively shunted junction (RSJ) equations describing a ladder array of overdamped, critical-current disordered Josephson junctions that are current biased along the rungs of the ladder can be mapped onto a Kuramoto model with nearest neighbor, sinusoidal couplings. This result is obtained by an averaging method, in which the fast dynamics of the RSJ equations are integrated out, leaving the dynamics which describe the time scale over which neighboring junctions along the rungs of the ladder phase and frequency synchronize. We quantify the degree of frequency synchronization of the rung junctions by calculating the standard deviation of their time-averaged voltages, sigma(omega), and the phase synchronization is quantified by calculating the time average of the modulus of the Kuramoto order parameter, <|r|>. We test the results of our averaging process by comparing the values of sigma(omega) and <|r|> for the original RSJ equations and our averaged equations. We find excellent agreement for dc bias currents of I(B)/ greater, similar 3, where is the average critical current of the rung junctions, and critical current disorders of up to 10%. We also study the effects of thermal noise on the synchronization properties of the overdamped ladder. Finally, we find that including the effects of junction capacitance can lead to a discontinuous synchronization transition as the strength of the coupling between neighboring junctions is smoothly varied. PMID- 12636786 TI - Information capacity and pattern formation in a tent map network featuring statistical periodicity. AB - We provide quantitative support to the observation that lattices of coupled maps are "efficient" information coding devices. It has been suggested recently that lattices of coupled maps may provide a model of information coding in the nervous system because of their ability to create structured and stimulus-dependent activity patterns which have the potential to be used for storing information. In this paper, we give an upper bound to the effective number of patterns that can be used to store information in the lattice by evaluating numerically its information capacity or information rate as a function of the coupling strength between the maps. We also estimate the time taken by the lattice to establish a limiting activity pattern. PMID- 12636787 TI - Parametrically forced surface wave with a nonmonotonic dispersion relation. AB - Surface wave patterns that arise in a mechanically driven ferrofluid system under constant magnetic field are investigated (1) to find out what kind of spatial patterns emerge when the system acquires a nonmonotonic dispersion relation and (2) to compare its surface wave patterns with those produced in the magnetically driven system studied earlier. As the strength of the applied magnetic field increases, the initial subharmonic square lattice formed by the Faraday instability first transforms to rolls, then becomes a rhomboid lattice. The rolls and the rhomboid lattice are found to coexist for a finite range of parameter space forming patterns with mixed domains. Possible underlying mechanisms for the observed rhomboid lattice is discussed. None of the diverse superlattices observed in the magnetically driven ferrofluid system appears in the mechanically driven system studied here. PMID- 12636788 TI - Two-degree-of-freedom Hamiltonian for the time-symmetric two-body problem of the relativistic action-at-a-distance electrodynamics. AB - We find a two-degree-of-freedom Hamiltonian for the time-symmetric problem of straight line motion of two electrons in direct relativistic interaction. This time-symmetric dynamical system appeared 100 years ago and it was popularized in the 1940s by the work of Wheeler and Feynman in electrodynamics, which was left incomplete due to the lack of a Hamiltonian description. The form of our Hamiltonian is such that the action of a Lorentz transformation is explicitly described by a canonical transformation (with rescaling of the evolution parameter). The method is closed and defines the Hamitonian in implicit form without power expansions. We outline the method with an emphasis on the physics of this complex conservative dynamical system. The Hamiltonian orbits are calculated numerically at low energies using a self-consistent steepest-descent method (a stable numerical method that chooses only the nonrunaway solution). The two-degree-of-freedom Hamiltonian suggests a simple prescription for the canonical quantization of the relativistic two-body problem. PMID- 12636789 TI - Dual synchronization of chaos in one-way coupled microchip lasers. AB - We experimentally demonstrate the dual synchronization of chaos in two pairs of Nd:YVO(4) microchip lasers in a one-way coupling configuration over one transmission channel. Dual synchronization is achieved when the optical frequency is matched between the corresponding pairs of lasers by using injection locking. We investigate the influence of optical injection from the two master lasers to one slave laser, and found that the dual synchronization is observed when the injection locking is achieved between either of the master lasers and the slave laser. Under the condition of the injection locking between both of the master lasers and the slave laser, the injection locking is alternately achieved and the accuracy of dual synchronization is degraded. We also confirm these results by numerical calculation. PMID- 12636790 TI - Wave-particle dualism of spiral waves dynamics. AB - We demonstrate and explain a wave-particle dualism of such classical macroscopic phenomena as spiral waves in active media. That means although spiral waves appear as nonlocal processes involving the whole medium, they respond to small perturbations as effectively localized entities. The dualism appears as an emergent property of a nonlinear field and is mathematically expressed in terms of the spiral waves response functions, which are essentially nonzero only in the vicinity of the spiral wave core. Knowledge of the response functions allows quantitatively accurate prediction of the spiral wave drift due to small perturbations of any nature, which makes them as fundamental characteristics for spiral waves as mass is for the condensed matter. PMID- 12636791 TI - Improvement of time-delayed feedback control by periodic modulation: analytical theory of Floquet mode control scheme. AB - We investigate time-delayed feedback control schemes which are based on the unstable modes of the target state, to stabilize unstable periodic orbits. The periodic time dependence of these modes introduces an external time scale in the control process. Phase shifts that develop between these modes and the controlled periodic orbit may lead to a huge increase of the control performance. We illustrate such a feature on a nonlinear reaction diffusion system with global coupling and give a detailed investigation for the Rossler model. In addition we provide the analytical explanation for the observed control features. PMID- 12636792 TI - Detecting generalized synchrony: an improved approach. AB - We examine some of the difficulties involved in detecting generalized synchrony (GS) in systems that exhibit noninvertibility and/or wrinkling. These latter features severely hinder identification of GS by conventional techniques. It is shown that it is possible to greatly improve detection by reducing the pseudofalse neighbors effects. Here we propose the delta(p)-neighbor method to overcome the noninvertibility effect and the delta(p,q) method to detect GS in systems with wrinkled structures. PMID- 12636793 TI - Diffusive transport and self-consistent dynamics in coupled maps. AB - The study of diffusion in Hamiltonian systems has been a problem of interest for a number of years. In this paper we explore the influence of self-consistency on the diffusion properties of systems described by coupled symplectic maps. Self consistency, i.e., the backinfluence of the transported quantity on the velocity field of the driving flow, despite of its critical importance, is usually overlooked in the description of realistic systems, for example, in plasma physics. We propose a class of self-consistent models consisting of an ensemble of maps globally coupled through a mean field. Depending on the kind of coupling, two different general types of self-consistent maps are considered: maps coupled to the field only through the phase, and fully coupled maps, i.e., through the phase and the amplitude of the external field. The analogies and differences of the diffusion properties of these two kinds of maps are discussed in detail. PMID- 12636794 TI - Simple potential-flow model of Rayleigh-Taylor and Richtmyer-Meshkov instabilities for all density ratios. AB - We generalize the Layzer-type model for unstable interfaces to the system of arbitrary density ratio. The predictions from the generalized model for bubble growth rates of Rayleigh-Taylor (RT) and Richtmyer-Meshkov (RM) instabilities are in good agreement with numerical results. We present the theoretical prediction for asymptotic growth rates for RT and RM bubbles for finite density ratios in two and three dimensions. PMID- 12636795 TI - Surface roughness and effective stick-slip motion. AB - The effect of random surface roughness on hydrodynamics of viscous incompressible liquid is discussed. When the hydrodynamic decay length (the viscous wave penetration depth) is larger than the correlation radius (size) of random surface inhomogeneities, it is possible to replace a random rough surface by effective stick-slip boundary conditions on a flat surface with two constants: the stick slip length and the renormalization of viscosity near the boundary. The stick slip length and the renormalization coefficient are expressed explicitly via the correlation function of random surface inhomogeneities. The stick-slip length is always negative and the effective change of viscosity near the surface is positive signifying the effective average hampering of the hydrodynamic flows by the rough surface (stick rather than slip motion). A simple hydrodynamic model illustrates general hydrodynamic results. The effective boundary parameters are analyzed numerically for Gaussian, power-law and exponentially decaying correlators with various indices. The maximum on the frequency dependence of the dissipation allows one to extract the correlation radius (characteristic size) of the surface inhomogeneities directly from, for example, experiments with torsional quartz oscillators. PMID- 12636796 TI - Stable nonspherical bubble collapse including period doubling in sonoluminescence. AB - We present observations of stable spherical symmetry broken states in single bubble sonoluminescence including observations of period doubled states. States observed involve both spatially oriented states and states with a tumbling symmetry axis. The observations are made using a fiber based four-channel correlation scheme. The measurements are made both with and without narrow band optical filters. The symmetry broken states are seen in all cases even using a 650+/-40-nm filter. This fact may be used to distinguish between different theories for the light emission. Prior to the measurements reported here, theoretical attempts to explain observations of period doubling bifurcation phenomena in single bubble sonoluminescence were centered on radially bifurcated collapses. The present experiments show unequivocally that the observations are primarily a result of breaking the spherical symmetry in the bubble collapse. Period doubling will at most show up as secondary effects in the total light output, if at all. PMID- 12636797 TI - Large-scale features of rotating forced turbulence. AB - Large-scale features of a randomly isotropically forced incompressible and unbounded rotating fluid are examined in perturbation theory. At first order in both the random force amplitude and the angular velocity, we find two types of modifications to the fluid equation of motion. The first correction transforms the molecular shear viscosity into a (rotation independent) effective viscosity. The second perturbative correction leads to a new large scale nondissipative force proportional to the fluid angular velocity in the slow rotation regime. This effective force does no net work and alters the dispersion relation of inertial waves propagating in the fluid. Both dynamically generated corrections can be identified with certain components of the most general axisymmetric "viscosity tensor" for a Newtonian fluid. PMID- 12636798 TI - Intermittency and exponent field dynamics in developed turbulence. AB - Spatiotemporal dynamics of intermittency in association with coarse-grained energy-dissipation rate fluctuations is discussed. This is done first by phenomenologically constructing the probability density for exponent field fluctuations that is introduced to characterize the energy-dissipation rate field, and then by proposing the Langevin dynamics derived with the projection operator method on the basis of the Navier-Stokes equation. With a Gaussian approximation for exponent fluctuations, spatiotemporal correlation functions for coarse-grained energy-dissipation rate fluctuations are explicitly obtained. PMID- 12636799 TI - Competition of spiral-defect chaos and rolls in Rayleigh-Benard convection under shear flow. AB - Computer simulations of domain coarsening of Rayleigh-Benard convective patterns under horizontal shear flow are carried out. The model calculations reported here explicitly include the hydrodynamic interaction of the order parameter field and provide a description of the spiral-defect chaos which competes with the roll pattern. We observe shear banding at moderate strain rates. PMID- 12636800 TI - High initial amplitude and high Mach number effects on the evolution of the single-mode Richtmyer-Meshkov instability. AB - Effects of high-Mach numbers and high initial amplitudes on the evolution of the single-mode Richtmyer-Meshkov shock-wave induced hydrodynamic instability are studied using theoretical models, experiments, and numerical simulations. Two regimes in which there is a significant deviation from the linear dependence of the initial velocity on the initial perturbation amplitude are defined and characterized. In one, the observed reduction of the initial velocity is primarily due to large initial amplitudes. This effect is accurately modeled by a vorticity deposition model, quantifying both the effect of the initial perturbation amplitude and the exact shape of the interface. In the other, the reduction is dominated by the proximity of the shock wave to the interface. This effect is modeled by a modified incompressible model where the shock wave is mimicked by a moving bounding wall. These results are supplemented with high initial amplitude Mach 1.2 shock-tube experiments, enabling separation of the two effects. It is shown that in most of the previous experiments, the observed reduction is predominantly due to the effect of high initial amplitudes. PMID- 12636801 TI - Experiment and simulation of laminar and turbulent ferrofluid pipe flow in an oscillating magnetic field. AB - Laminar and turbulent pipe flow of a ferrofluid with an imposed linearly polarized, oscillating, magnetic field is examined here. Experimental results show a fractional pressure drop dependence on flow rate, magnetic field strength, and oscillation frequency. Calculations are presented, which show that ferrofluid theory can explain the flow phenomena in laminar and turbulent pipe flow. The model requires an initial fit of key parameters but then shows predictive capability in both laminar and turbulent flow. Simulation results are found to be essentially independent of the spin boundary condition due to an approximation of spin viscosity that is very small. A low Reynolds number k-epsilon model is used to model the turbulent pipe flow. PMID- 12636802 TI - Lattice Boltzmann simulation of fluid flow in fracture networks with rough, self affine surfaces. AB - Using the lattice Boltzmann method, we study fluid flow in a two-dimensional (2D) model of fracture network of rock. Each fracture in a square network is represented by a 2D channel with rough, self-affine internal surfaces. Various parameters of the model, such as the connectivity and the apertures of the fractures, the roughness profile of their surface, as well as the Reynolds number for flow of the fluid, are systematically varied in order to assess their effect on the effective permeability of the fracture network. The distribution of the fractures' apertures is approximated well by a log-normal distribution, which is consistent with experimental data. Due to the roughness of the fractures' surfaces, and the finite size of the networks that can be used in the simulations, the fracture network is anisotropic. The anisotropy increases as the connectivity of the network decreases and approaches the percolation threshold. The effective permeability K of the network follows the power law K approximately (beta), where is the average aperture of the fractures in the network and the exponent beta may depend on the roughness exponent. A crossover from linear to nonlinear flow regime is obtained at a Reynolds number Re approximately O(1), but the precise numerical value of the crossover Re depends on the roughness of the fractures' surfaces. PMID- 12636803 TI - Stabilization of hydrodynamic flows by small viscosity variations. AB - Motivated by the large effect of turbulent drag reduction by minute concentrations of polymers, we study the effects of a weakly space-dependent viscosity on the stability of hydrodynamic flows. In a recent paper [Phys. Rev. Lett. 87, 174501, (2001)], we exposed the crucial role played by a localized region where the energy of fluctuations is produced by interactions with the mean flow (the "critical layer"). We showed that a layer of a weakly space-dependent viscosity placed near the critical layer can have a very large stabilizing effect on hydrodynamic fluctuations, retarding significantly the onset of turbulence. In this paper we extend these observations in two directions: first we show that the strong stabilization of the primary instability is also obtained when the viscosity profile is realistic (inferred from simulations of turbulent flows with a small concentration of polymers). Second, we analyze the secondary instability (around the time-dependent primary instability) and find similar strong stabilization. Since the secondary instability develops around a time-dependent solution and is three dimensional, this brings us closer to the turbulent case. We reiterate that the large effect is not due to a modified dissipation (as is assumed in some theories of drag reduction), but due to reduced energy intake from the mean flow to the fluctuations. We propose that similar physics act in turbulent drag reduction. PMID- 12636804 TI - Dynamics of low-energy helium vapor pulses. AB - We report results of experiments in which pulses of helium vapor are produced by a current pulse in a chromium film covered with superfluid helium at around 0.3 K. The pulses were detected by a titanium bolometer operating at 0.47 K. The shape of the detected signal is a strong function of the power of the initiating current pulse. For low powers the signal from a single current pulse also contains a single peak, but for higher powers, a single current pulse produces two and then at the highest powers, three peak signals. To analyze the origin of these phenomena we report results of hybrid gas-dynamics and hydrodynamics simulations, which demonstrate that the signals arise from shock waves formed in the vapor. The shock waves form due to the presence of a gradient in the small ambient background of helium vapor in the chamber and are extremely sensitive to the pulse power. PMID- 12636805 TI - Scaling exponents in anisotropic hydrodynamic turbulence. AB - In anisotropic turbulence, the correlation functions are decomposed in the irreducible representations of the SO(3) symmetry group (with different "angular momenta" l). For different values of l, the second-order correlation function is characterized by different scaling exponents zeta(2)(l). In this paper, we compute these scaling exponents in a closure approximation. By linearizing the closure equations in small anisotropy we set up a linear operator and find its zero modes in the inertial interval of scales. Thus the scaling exponents in each l sector follow from solvability condition, and are not determined by dimensional analysis. The main result of our calculation is that the scaling exponents zeta(2)(l) form a strictly increasing spectrum at least until l=6, guaranteeing that the effects of anisotropy decay as power laws when the scale of observation diminishes. The results of our calculations are compared to available experiments and simulations. PMID- 12636806 TI - Mode-coupling approach to non-Newtonian Hele-Shaw flow. AB - The Saffman-Taylor viscous fingering problem is investigated for the displacement of a non-Newtonian fluid by a Newtonian one in a radial Hele-Shaw cell. We execute a mode-coupling approach to the problem and examine the morphology of the fluid-fluid interface in the weak shear limit. A differential equation describing the early nonlinear evolution of the interface modes is derived in detail. Owing to vorticity arising from our modified Darcy's law, we introduce a vector potential for the velocity in contrast to the conventional scalar potential. Our analytical results address how mode-coupling dynamics relates to tip splitting and sidebranching in both shear-thinning and shear-thickening cases. The development of non-Newtonian interfacial patterns in rectangular Hele-Shaw cells is also analyzed. PMID- 12636807 TI - Anomalous transport of particle tracers in multidimensional cellular flows. AB - Advection of tracers is studied numerically in time-dependent, two-dimensional cellular flows and a time-independent, three-dimensional cellular flow field. Tracers in these flows follow trajectories that are either periodic or chaotic and mimic correlated Levy flights. The probability density function of displacements for particles in the ordered regions of the flow follows a classical Gaussian dispersion process. The particle trajectories in the chaotic regions of the flow exhibit anomalous diffusion and the probability density function of displacements is well modeled by a time-fractional diffusion equation of order alpha. The overall process of particle dispersion is found to be controlled mainly by the chaotic regions within the flow field. From the perspective of Lagrangian dynamics our results indicate that the advection of particles in flow fields prone to exhibit chaotic advection is a combination of both classical, i.e., Gaussian, behavior and anomalous, i.e., non-Gaussian, diffusion. PMID- 12636808 TI - Simulations of two-dimensional turbulent convection in a density-stratified fluid. AB - High resolution computer simulations of two-dimensional convection using the anelastic approximation are presented. These calculations span Rayleigh numbers from 10(8)-10(12) for Prandtl number equal to unity, with the fluid density decreasing by a factor of 12 from the bottom to the top of the convection region. This range covers several decades in the "hard" turbulent regime. While many studies of this sort have been conducted for the Boussinesq approximation (i.e., no density stratification), we use the anelastic approximation with a significant density stratification in this turbulent regime. The convection is dominated by a large-scale coherent flow composed of ascending and descending superplumes. We find a power law exponent of 0.28 for the Nusselt-Rayleigh number scaling and a power law with exponent of 0.50 for the Reynolds-Rayleigh number scaling for the entire parameter space studied. These values are very similar to those determined experimentally and analytically for convection with no density stratification. PMID- 12636809 TI - Roles of convection, pressure, and dissipation in three-dimensional turbulence. AB - We investigated the roles of the convection, pressure, and dissipation terms in the equation for the longitudinal velocity increment, with the help of the DNS data on 1024(3). The pressure screens the convection growing as the intensity of fluctuation increases. The dissipation term is found to make no direct contribution to the fourth order structure function; the structure functions of order not less than 5 are affected by the dissipative structure as far as the scaling is concerned. A reason is also given for the observation by Stolovitzky et al. [Phys. Rev. E 48, R3217 (1993)] that the scaling of the (2m)th order structure function is more similar to that of the (2m-1)th order structure function than that of (2m+1)th order structure function. PMID- 12636810 TI - Spectral intermode coupling in a model of isotropic turbulence. AB - We investigate the nonlinear coupling between the so-called explicit modes, identified with wave numbers k such that 0< or =k< or =k(c), and implicit modes, defined such that k(c)< or =k< or =k(max). Here k(c) is an arbitrarily chosen cutoff wave number and k(max) is the ultraviolet cutoff as determined by viscous damping. The stresses arising from the nonlinearity in the Navier-Stokes equations are categorized as "implicit-implicit" (or "Reynolds") and "explicit implicit" (or "cross"). These arise from dynamic coupling between different regions of wave number space. Their respective effects on momentum, kinetic energy, and energy flux are assessed. The analysis is based on a model system comprising the Navier-Stokes equations and the Edwards-Fokker-Planck energy equation [S. F. Edwards, J. Fluid Mech. 18, 239 (1964)] which is known to retain all the symmetries of homogeneous, isotropic turbulence. The Reynolds stress is found to be responsible for long-range energy transfers. It can be represented by an effective viscosity and is mainly determined by dynamical friction. The cross term is more complicated, involving both diffusive and frictional effects. For long-range coupling it can be expressed as a modification of the effective viscosity, while for short-range coupling it may be modeled on the assumption that implicit scales are slaved to explicit scales. Thus, both the random and coherent aspects of intermode coupling in turbulent flows are relevant in the cross term. The imposition of a continuity requirement on energy transfer leads to a new parametrization that represents the effect of absent modes in a truncated spectral simulation, and takes into account the phase-coupling (coherent) effects, as well as the usual viscositylike (random) effects. PMID- 12636811 TI - Spiral and Taylor vortex fronts and pulses in axial through flow. AB - The influence of an axial through flow on the spatiotemporal growth behavior of different vortex structures in the Taylor-Couette system with radius ratio eta=0.5 is determined. The Navier-Stokes equations (NSE) linearized around the basic Couette-Poiseuille flow are solved numerically with a shooting method in a wide range of through flow strengths Re and different rates of co-rotating and counter-rotating cylinders for toroidally closed vortices with azimuthal wave number m=0 and for spiral vortex flow with m=+/-1. For each of these three different vortex varieties we have investigated (i) axially extended vortex structures, (ii) axially localized vortex pulses, and (iii) vortex fronts. The complex dispersion relations of the linearized NSE for vortex modes with the three different m are evaluated for real axial wave numbers for (i) and over the plane of complex axial wave numbers for (ii) and (iii). We have also determined the Ginzburg-Landau amplitude equation (GLE) approximation in order to analyze its predictions for the vortex structures (ii) and (iii). Critical bifurcation thresholds for extended vortex structures are evaluated. The boundaries between absolute and convective instability of the basic state for vortex pulses are determined with a saddle-point analysis of the dispersion relations. Fit parameters for power-law expansions of the boundaries up to Re4 are listed in two tables. Finally, the linearly selected front behavior of growing vortex structures is investigated using saddle-point analyses of the dispersion relations of NSE and GLE. For the two front intensity profiles (increasing in positive or negative axial direction) we have determined front velocities, axial growth rates, and the wave numbers and frequencies of the unfolding vortex patterns with azimuthal wave numbers m=0,+/-1, respectively. PMID- 12636812 TI - Explicit expressions for the evolution of single-mode Rayleigh-Taylor and Richtmyer-Meshkov instabilities at arbitrary Atwood numbers. AB - We present explicit analytic expressions for the evolution of the bubble amplitude in Rayleigh-Taylor (RT) and Richtmyer-Meshkov RM instabilities. These expressions are valid from the linear to the nonlinear regime and for arbitrary Atwood number A. Our method is to convert from the linear to the nonlinear solution at a specific value eta* of the amplitude for which explicit analytic expressions have been given previously for A=1 [K. O. Mikaelian, Phys. Rev. Lett. 80, 508 (1998)]. By analyzing a recent extension of Layzer's theory to arbitrary A [V. N. Goncharov, Phys. Rev. Lett. 88, 134502 (2002)], we find a simple transformation that generalizes our solutions to arbitrary A. We compare this model with another explicit model attributed to Fermi and with numerical simulations. Fermi's model agrees with numerical simulations for the RT case but its extension to the RM case disagrees with simulations. The model proposed here agrees with hydrocode calculations for both RT and RM instabilities. PMID- 12636813 TI - Alternative method to deduce bubble dynamics in single-bubble sonoluminescence experiments. AB - In this paper we present an experimental approach that allows to deduce the important dynamical parameters of single sonoluminescing bubbles (pressure amplitude, ambient radius, radius-time curve). The technique is based on a few previously confirmed theoretical assumptions and requires the knowledge of quantities such as the amplitude of the electric excitation and the phase of the flashes in the acoustic period. These quantities are easily measurable by a digital oscilloscope, avoiding the cost of the expensive lasers or ultrafast cameras of previous methods. We show the technique in a particular example and compare the results with conventional Mie scattering. We find that within the experimental uncertainties these two techniques provide similar results. PMID- 12636814 TI - Effect of rotation on a developed turbulent stratified convection: the hydrodynamic helicity, the alpha effect, and the effective drift velocity. AB - An effect of rotation on a developed turbulent stratified convection is studied. Dependences of the hydrodynamic helicity, the alpha tensor, and the effective drift velocity of the mean magnetic field on the rate of rotation and an anisotropy of turbulent convection are found. It is shown that in an anisotropic turbulent convection the alpha effect can change its sign depending on the rate of rotation. The evolution of the alpha effect is much more complicated than that of the hydrodynamic helicity in an anisotropic turbulent convection of a rotating fluid. Different properties of the effective drift velocity of the mean magnetic field in a rotating turbulent convection are found: (i) a poloidal effective drift velocity can be diamagnetic or paramagnetic depending on the rate of rotation; (ii) there is a difference in the effective drift velocities for the toroidal and poloidal magnetic fields; (iii) a toroidal effective drift velocity can play a role of an additional differential rotation. The above effects and an effect of a nonzero divergence of the effective drift velocity of the toroidal magnetic field on a magnetic dynamo in a developed turbulent stratified convection of a rotating fluid are studied. Astrophysical applications of the obtained results are discussed. PMID- 12636815 TI - Convective heat transport in a rotating fluid layer of infinite Prandtl number: optimum fields and upper bounds on Nusselt number. AB - By means of the Howard-Busse method of the optimum theory of turbulence we investigate numerically upper bounds on convective heat transport for the case of infinite fluid layer with stress-free vertical boundaries rotating about a vertical axis. We discuss the case of infinite Prandtl number, 1-alpha solution of the obtained variational problem and optimum fields possessing internal, intermediate, and boundary layers. We investigate regions of Rayleigh and Taylor numbers R and Ta, where no analytical bounds can be derived, and compare the analytical and numerical bounds for these regions of R and Ta where such comparison is possible. The increasing rotation has a different influence on the rescaled optimum fields of velocity w(1), temperature theta(1) and the vertical component of the vorticity f(1). The increasing Ta for fixed R leads to vanishing of the boundary layers of w(1) and theta(1). Opposite to this, the increasing Ta leads first to a formation of boundary layers of the field f(1) but further increasing the rotation causes vanishing of these boundary layers. We obtain optimum profiles of the horizontal averaged total temperature field which could be used as hints for construction of the background fields when applying Doering Constantin method to the problems of rotating convection. The wave number alpha(1) corresponding to the optimum fields follows the asymptotic relationship alpha(1)=(R/5)(1/4) for intermediate Rayleigh numbers. However, when R becomes large with respect to Ta, after a transition region, the power law for alpha(1) becomes close to the power law for the case without rotation. The Nusselt number Nu is close to the nonrotational bound 0.32R(1/3) for the case of large R and small Ta. Nu decreases with increasing Taylor number. Thus, the upper bounds reflect the tendency of inhibiting thermal convection by increasing rotation for a fixed Rayleigh number. For the regions of Rayleigh and Taylor numbers where the numerical and asymptotic bounds on Nu can be compared, the numerical bounds are about 70% lower than the asymptotic bounds. PMID- 12636816 TI - Contributions to the theory of a two-scale homogeneous dynamo experiment. AB - The principle of the two-scale dynamo experiment at the Forschungszentrum Karlsruhe is closely related to that of the Roberts dynamo working with a simple fluid flow which is, with respect to proper Cartesian coordinates x, y, and z, periodic in x and y and independent of z. A modified Roberts dynamo problem is considered with a flow more similar to that in the experimental device. Solutions are calculated numerically, and on this basis an estimate of the excitation condition of the experimental dynamo is given. The modified Roberts dynamo problem is also considered in the framework of the mean-field dynamo theory, in which the crucial induction effect of the fluid motion is an anisotropic alpha effect. Numerical results are given for the dependence of the mean-field coefficients on the fluid flow rates. The excitation condition of the dynamo is also discussed within this framework. The behavior of the dynamo in the nonlinear regime, i.e., with backreaction of the magnetic field on the fluid flow, depends on the effect of the Lorentz force on the flow rates. The quantities determining this effect are calculated numerically. The results for the mean-field coefficients and the quantities describing the backreaction provide corrections to earlier results, which were obtained under simplifying assumptions. PMID- 12636817 TI - Two-dimensional solitons and vortices in normal and anomalous dispersive media. AB - We study solitons and vortices described by the (2+1)-dimensional fourth-order generalized nonlinear Schrodinger equation with cubic-quintic nonlinearity. Necessary conditions for the existence of such structures are investigated analytically using conservation laws and asymptotic behavior of localized solutions. We derive the generalized virial relation, which describes the combined influence of linear and nonlinear effects on the evolution of the wave packet envelope. By means of refined variational analysis, we predict the main features of steady soliton solutions, which have been shown to be in good agreement with our numerical results. Soliton and vortex stability is investigated by linear analysis and direct numerical simulations. We show that stable bright solitons exist in nonlinear Kerr media both in anomalous and normal dispersive regimes, even if only the fourth-order dispersive effect is taken into account. Vortices occur robust with respect to symmetry-breaking azimuthal instability only in the presence of additional defocusing quintic nonlinearity in the strongly nonlinear regime. We apply our results to the theoretical explanation of whistler self-induced waveguide propagation in plasmas, and discuss possible applications to light beam propagation in cubic-quintic optical materials and to solitons in two-dimensional molecular systems. PMID- 12636818 TI - Origin of protons accelerated by an intense laser and the dependence of their energy on the plasma density. AB - We study the high-energy (1-4 MeV) proton production from a slab plasma irradiated by a ultrashort high-power laser. In our 2.5-dimensional particle-in cell simulations, a p-polarized laser beam of 1.6 x 10(19) W/cm(2), 300 fs, lambda(L)=1.053 microm, illuminates a slab plasma normally; the slab plasma consists of a hydrogen plasma, and the target plasma thickness and the laser spot size are 2.5lambda(L) and 5lambda(L), respectively. The simulation results show that an emitted proton energy depends on the slab plasma density, and three kinds of high-energy proton beams are generated at the target plasma surfaces: one kind of the proton beams is produced at the laser-illuminated target surface and accelerated to the same laser-incident side. The second is generated at the target surface opposite to the laser-illuminated target surface and is accelerated outward on the same side. The third is generated at the laser illuminated target surface and accelerated to the opposite side while passing through the target plasma. The simulations also show a mechanism of proton accelerations. In an overdense plasma, laser energy goes to energies of hot electrons and magnetic fields in part; the electrons oscillate around the slab plasma so that a static electric field is generated and consequently protons are extracted. The magnetic field generated in the slab plasma exists longer and heats up the plasma electrons to sustain the static electric field even after the laser termination. PMID- 12636819 TI - Saturation and postsaturation phenomena of Rayleigh-Taylor instability with adjacent modes. AB - A weakly nonlinear theory has been developed for the classical Rayleigh-Taylor instability with a finite bandwidth taken into account self-consistently. The theory includes up to third order nonlinearity, which results in the saturation of linear growth and determines subsequent weakly nonlinear growth. Analytical results are shown to agree fairly well with two-dimensional hydrodynamic simulations. There are generally many local peaks of a perturbation with a finite bandwidth due to the interference of modes. Since a local amplitude is determined from phases among the modes as well as the bandwidth, we have investigated an onset of the linear growth saturation and the subsequent weakly nonlinear growth for different bandwidths and phases. It is shown that the saturation of the linear growth occurs locally, i.e., each of the local maximum amplitudes (LMAs) grows exponentially until it reaches almost the same saturation amplitude. In the random phase case, the root mean square amplitude thus saturates with almost the same amplitude as the LMA, after most of the LMAs have saturated. The saturation amplitude of the LMA is found to be independent of the bandwidth and depends on the Atwood number. We derive a formula of the saturation amplitude of modes based on the results obtained, and discuss its relation with Haan's formula [Phys. Rev. A 39, 5812 (1989)]. The LMAs grow linearly in time after the saturation and their speeds are approximated by the product of the linear growth rate and the saturation amplitude. We investigate the Atwood number dependence of both the saturation amplitude and the weakly nonlinear growth. PMID- 12636820 TI - Vlasov-Maxwell kinetic simulations of radio-frequency-driven ion flows in magnetized plasmas. AB - The generation of a coherent ion flow due to the injection in a plasma of a purely electrostatic wave of finite amplitude, propagating at right angle with the ambient uniform magnetic field, is investigated making use of a kinetic code which solves the fully nonlinear Vlasov equations for electrons and ions, coupled with the Maxwell equations, in one spatial and two velocity dimensions. A uniformly magnetized slab plasma is considered. The wave frequency is assumed in the range of the fourth harmonic of the ion cyclotron frequency, and the wave vector is chosen in order to model the propagation of an ion Bernstein wave. The computation of the first-order moment of the ion distribution function shows that indeed a quasistationary transverse average ion drift velocity is produced. The time evolution of the ion distribution function undergoes a "resonant" interaction of Cherenkov type, even if the plasma ions are magnetized (omega(ci)/omega(pi) approximately 0.5). During the wave-plasma interaction, the electron distribution function remains Gaussian-like, while increasing its energy content. PMID- 12636821 TI - Magnetic line trapping and effective transport in stochastic magnetic fields. AB - The transport of collisional particles in stochastic magnetic fields is studied using the decorrelation trajectory method. The nonlinear effect of magnetic line trapping is considered together with particle collisions. The running diffusion coefficient is determined for arbitrary values of the statistical parameters of the stochastic magnetic field and of the collisional velocity. The effect of the magnetic line trapping is determined. New anomalous diffusion regimes are found. PMID- 12636822 TI - Quasilinear theory of Cherenkov-drift instability. AB - We examine linear and quasiliner stages of Cherenkov-drift instability developed in the relativistic magnetized electron-positron plasma penetrated by ultrarelativistic beam of electrons (or positrons). The plasma flow is streaming along the slightly curved magnetic field lines. In this case, the curvature drift of beam particles plays a decisive role in the development of the instability. A quasilinear relaxation of Cherenkov-drift instability leads to diffusion of resonant particles in momenta space. The expressions for diffusion coefficients of Cherenkov-drift instability are obtained. The numerical estimations are carried out for the parameters of relativistic magnetized plasma of pulsar magnetospheres providing a test of validity of the approximations used in our approach. PMID- 12636823 TI - Propagation of hot electrons through high-density plasmas. AB - Propagation of hot electrons through high-density plasmas generated by femtosecond laser pulses is investigated using three types of target configurations: Al-coated glass, Al and glass separated by a vacuum gap, and Al foil alone. Collimated ionization tracks lasting for 60 ps and extending 150-300 microm in length and 8 microm in cross section are observed via optical probing. For the Al-foil-alone target, a narrow plasma jet is formed at the rear surface in line with the laser. The collimation of the hot electrons may be attributed to a strong self-generated magnetic field in the target. PMID- 12636824 TI - Advanced spectroscopic analysis of 0.8-1.0-MA Mo x pinches and the influence of plasma electron beams on L-shell spectra of Mo ions. AB - This paper presents a detailed investigation of the temporal, spatial, and spectroscopic properties of L-shell radiation from 0.8 to 1.0 MA Mo x pinches. Time-resolved measurements of x-ray radiation and both time-gated and time integrated spectra and pinhole images are presented and analyzed. High-current x pinches are found to have complex spatial and temporal structures. A collisional radiative kinetic model has been developed and used to interpret L-shell Mo spectra. The model includes the ground state of every ionization stage of Mo and detailed structure for the O-, F-, Ne-, Na-, and Mg-like ionization stages. Hot electron beams generated by current-carrying electrons in the x pinch are modeled by a non-Maxwellian electron distribution function and have significant influence on L-shell spectra. The results of 20 Mo x-pinch shots with wire diameters from 24 to 62 microm have been modeled. Overall, the modeled spectra fit the experimental spectra well and indicate for time-integrated spectra electron densities between 2 x 10(21) and 2 x 10(22) cm(-3), electron temperatures between 700 and 850 eV, and hot electron fractions between 3% and 7%. Time-gated spectra exhibit wide variations in temperature and density of plasma hot spots during the same discharge. PMID- 12636825 TI - Interchange mode in the presence of dust. AB - The linear and nonlinear development of an electrostatic interchange mode which involves a magnetized nonuniform electron-ion fluid in the presence of nonuniform static charged dust grains is investigated. The charge on grains is taken as spatially dependent, and the consequences of that condition are investigated. It is shown that standardly accepted stabilization of the interchange mode in the presence of negatively charged grains can be violated due to the spatial dependence of the charge on grains. Also, the ion drift, which is caused by the action of a gravity term perpendicular to the magnetic field lines, is taken as nonuniform as a result of the magnetic field nonuniformity, and it is shown that due to such a nonuniformity the instability condition can be significantly modified. In the nonlinear regime several types of coherent stationary vortex structures are found: namely, dipolar and tripolar vortices and vortex chains. The dipolar vortex is found to propagate in the direction of the ion drift, while the tripole and vortex chains are carried by the drift flow. The spatial dependence of these structures is determined by parameters describing the nonuniformity of the equilibrium plasma. PMID- 12636826 TI - Energy loss of ions in a magnetized plasma: conformity between linear response and binary collision treatments. AB - The energy loss of a heavy ion moving in a magnetized electron plasma is considered within the linear response (LR) and binary collision (BC) treatments with the purpose to look for a connection between these two models. These two complementary approaches yield close results if no magnetic field is present, but there develop discrepancies with growing magnetic field at ion velocities that are lower than, or comparable with, the thermal velocity of the electrons. We show that this is a peculiarity of the Coulomb interaction which requires cutoff procedures to account for its singularity at the origin and its infinite range. The cutoff procedures in the LR and BC treatments are different as the order of integrations in velocity and in ordinary (Fourier) spaces is reversed in both treatments. While BC involves a velocity average of Coulomb logarithms, there appear in LR Coulomb logarithms of velocity averaged cutoffs. The discrepancies between LR and BC vanish, except for small contributions of collective modes, for smoothened potentials that require no cutoffs. This is shown explicitly with the help of an improved BC in which the velocity transfer is treated up to second order in the interaction in Fourier space. PMID- 12636827 TI - Theoretical model of x-ray scattering as a dense matter probe. AB - We present analytical expressions for the dynamic structure factor, or form factor S(k,omega), which is the quantity describing the x-ray cross section from a dense plasma or a simple liquid. Our results, based on the random phase approximation for the treatment on the charged particle coupling, can be applied to describe scattering from either weakly coupled classical plasmas or degenerate electron liquids. Our form factor correctly reproduces the Compton energy down shift and the known Fermi-Dirac electron velocity distribution for S(k,omega) in the case of a cold degenerate plasma. The usual concept of scattering parameter is also reinterpreted for the degenerate case in order to include the effect of the Thomas-Fermi screening. The results shown in this work can be applied to interpreting x-ray scattering in warm dense plasmas occurring in inertial confinement fusion experiments or for the modeling of solid density matter found in the interior of planets. PMID- 12636828 TI - Random walk through fractal environments. AB - We analyze random walk through fractal environments, embedded in three dimensional, permeable space. Particles travel freely and are scattered off into random directions when they hit the fractal. The statistical distribution of the flight increments (i.e., of the displacements between two consecutive hittings) is analytically derived from a common, practical definition of fractal dimension, and it turns out to approximate quite well a power-law in the case where the dimension D(F) of the fractal is less than 2, there is though, always a finite rate of unaffected escape. Random walks through fractal sets with D(F)< or =2 can thus be considered as defective Levy walks. The distribution of jump increments for D(F)>2 is decaying exponentially. The diffusive behavior of the random walk is analyzed in the frame of continuous time random walk, which we generalize to include the case of defective distributions of walk increments. It is shown that the particles undergo anomalous, enhanced diffusion for D(F)<2, the diffusion is dominated by the finite escape rate. Diffusion for D(F)>2 is normal for large times, enhanced though for small and intermediate times. In particular, it follows that fractals generated by a particular class of self-organized criticality models give rise to enhanced diffusion. The analytical results are illustrated by Monte Carlo simulations. PMID- 12636829 TI - Neutral-plasma oscillations at zero temperature. AB - Cold plasma theory is used to calculate the response of an ultracold neutral plasma to an applied rf field. The free oscillation of the system has a continuous spectrum and an associated damped quasimode. This quasimode dominates the driven response and is resonant in the tail of the density distribution. Recent experiments used the plasma response to an applied rf field to determine the plasma density in an expanding ultracold plasma. The comparison between experiment and theory indicates that this method accurately determines the expansion velocity and underestimates the initial plasma density by a factor of 3 in weakly collisional plasmas. PMID- 12636830 TI - Direction-selective free expansion of laser-produced plasmas from planar targets. AB - Direction-selective expansion of laser-produced plasmas from planar slab targets of Al, Ni, Mo, and Ta are reported. Angular distributions of the particles emitted from the targets, produced by a 130 mJ, 5 nsec, Nd:YAG laser, were obtained by means of a retarding potential analyzer and a quartz crystal. It was observed that the angular distributions of the particles had mainly three characteristics. For a given laser energy and a given target element, the angular distribution showed more preferential focusing toward the target normal as the value of the focal spot size B increased. Second, for a given laser energy and a given focal spot size, the focusing was more pronounced toward the target normal as the atomic mass number of the target materials increased. Third, for a given energy, a given focal spot size and a given element, the particles with higher ionization states were much more focused toward the target normal. Our experimental results confirm the Monte Carlo simulation results of the earlier works taking into account collision and recombination processes. PMID- 12636831 TI - Electron acceleration by few-cycle laser pulses with single-wavelength spot size. AB - Generation of relativistic electrons from the interaction of a laser pulse with a high density plasma foil, accompanied by an underdense preplasma in front of it, has been studied with two-dimensional particle-in-cell (PIC) simulations for pulse durations comparable to a single cycle and for single-wavelength spot size. The electrons are accelerated predominantly in forward direction for a preplasma longer than the pulse length. Otherwise, both forward and backward electron accelerations occur. The primary mechanism responsible for electron acceleration is identified. Simulations show that the energy of the accelerated electrons has a maximum versus the pulse duration for relativistic laser intensities. The most effective electron acceleration takes place when the preplasma scale length is comparable to the pulse duration. Electron distribution functions have been found from PIC simulations. Their tails are well approximated by Maxwellian distributions with a hot temperature in the MeV range. PMID- 12636832 TI - Steady state of a fully detuned storage-ring free-electron laser. AB - This paper gives an analytical description of the stationary regime of a storage ring free-electron laser in the presence of the maximum detuning (compatible with the laser onset) between the laser pulse and the electron beam when they pass and interact in the optical cavity. In this condition, the conservation of the first moments of the laser intensity distribution allows one to express the peak gain of the light amplification process and the maximum detuning as a function of system parameters that are directly measurable. These theoretical results are compared with experiments performed on the Super-ACO free-electron laser. PMID- 12636833 TI - Relativistic nonlinear Thomson scattering as attosecond x-ray source. AB - Relativistic, nonlinear Thomson scattering by an electron of an intense laser field has been investigated by computer simulation. Under a laser field with a pulse duration of 20-fs full width at half maximum and an intensity of 10(20) W/cm(2), the motion of an electron is highly relativistic and generates an ultrashort radiation of 2 as with photon energies from 100 to 600 eV. An interesting modulated structure of the spectrum is observed and analyzed. A radiation produced by the zigzag motion of an electron under a linearly polarized laser has better characteristics than by a helical motion under a circularly polarized laser pulse in terms of an angular divergence and an energy spectrum. The effect of ion field in a plasma was also investigated, which shows that for a laser intensity of 10(20) W/cm(2), the ion field due to an ion density of up to 7 x 10(18) cm(-3) can be ignored during the laser pulse. PMID- 12636834 TI - Chaotic behavior of the radiation field in the magnetically insulated transmission line oscillator. AB - In terms of the nonlinear coupled equations that describe the evolution of the radiation field and electrons motion in the magnetically insulated transmission line oscillator, the nonlinear instabilities existing in the process of the interaction between the electron beam and field are analyzed. It is shown that the development of the instability leads to the appearance of the nonlinear behavior of the radiation field. (1) In the soft nonlinear regime in which the radiation field is characterized by the periodic limit cycle, the power spectrum is discrete and the sideband is not symmetric around the carrier frequency; in the hard nonlinear regime in which the radiation field is characterized by the chaotic oscillation, the power spectrum is continuous and the spectral components with larger amplitude distribute in the low frequency region. (2) The threshold current at which the limit cycle or chaotic oscillation starts occurring is a function of the detuning. The appearance of the nonlinear unsteady states may be accelerated and inhibited by controlling detuning. (3) The arising of the limit cycle oscillation and chaotic behavior of the radiation field is easier in a magnetically insulated transmission line oscillator than in a traveling wave tube amplifier. PMID- 12636835 TI - Resonant phenomena in classical dynamics of three-body Coulomb systems. AB - We consider dynamics of a planar three-body Coulomb system similar to a hydrogen molecular ion (heavy-light-heavy particles). The system has three degrees of freedom. In the limit of infinitely heavy nuclei the system is reduced to the famous two-center problem which is integrable. When masses of heavy particles are finite, one degree of freedom in the Hamiltonian system corresponds to slow nuclei motion, while other two degrees of freedom correspond to fast electron motion. The averaging method predicts that actions of "fast" motions of the system with frozen nuclei are approximate integrals of the full system (adiabatic invariants). However, during slow evolution of the "heavy" subsystem certain resonance conditions can be satisfied. We study the phenomena of capture into resonances and scattering on resonances which can lead to destruction of adiabatic invariance in the system. PMID- 12636836 TI - Influence of higher-order dispersion on modulational instability and pulse broadening of partially incoherent light. AB - The Wigner-Moyal equation for the Wigner distribution of a partially incoherent optical wave field propagating in dispersive and nonlinear media has been generalized to include the effects of both arbitrary order of dispersion and arbitrary nonlinearity. The theory predicts partial incoherence to enhance the pulse broadening during linear wave pulse propagation. Furthermore, an application of the theory to the modulational instability of constant amplitude waves shows how higher-order dispersion affects the instability growth rate. PMID- 12636837 TI - Modulation instability and solitons on a cw background in an optical fiber with higher-order effects. AB - We construct the Lax pair for a higher-order nonlinear Schrodinger equation that includes terms accounting for the third-order dispersion, the self-steepening effect, and the delayed nonlinear response effect. Two exact analytic solutions that describe (i) modulation instability and (ii) soliton propagation on a continuous wave background are obtained by using the Darboux transformation. In addition, we analyze the amplification-absorption and quintic nonlinearity effects on the second solution in the adiabatic approximation. PMID- 12636838 TI - Excitation of phase patterns and spatial solitons via two-frequency forcing of a 1:1 resonance. AB - We show that a self-oscillatory system, driven at two frequencies close to that of the unforced system (resonance 1:1), becomes phase locked and exhibits two equivalent stable states of opposite phases. For spatially extended systems this phase bistability results in patterns characteristic for real order parameter systems, such as phase domains, labyrinths, and phase spatial solitons. In variational cases, the phase-locking mechanism is interpreted as a result of the periodic "rocking" of the system potential. Rocking could be tested experimentally in lasers and in oscillatory chemical reactions. PMID- 12636839 TI - Minkowskian description of polarized light and polarizers. AB - A conventional Stokes description of polarized light is considered in a four dimensional Lorentzian space, developing a seminal idea of Paul Soleillet [Ann. Phys. (Paris) 12, 23 (1929)]. This provides a striking interpretation for the degree of polarization and the Stokes decomposition of light beams. Malus law and reciprocity theorems for polarizers are studied using this Lorentzian formalism. PMID- 12636840 TI - Discrete Ginzburg-Landau solitons. AB - We demonstrate that discrete solitons are possible in Ginzburg-Landau lattices. As a result of discreteness, we find that this system exhibits a host of features that have no counterpart whatsoever in either the continuous limit or in other conservative discrete models. PMID- 12636841 TI - Gap solitons and soliton trains in finite-sized two-dimensional periodic and quasiperiodic photonic crystals. AB - We demonstrate the existence of the gap solitons and soliton trains in finite sized two-dimensional periodic nonlinear photonic crystals by using the mutiple scattering approach with an iterative scheme. In 12-fold symmetric nonlinear quasicrystals, we also demonstrated the existence of symmetric, regular gap solitons, asymmetric single-soliton states, and two-solitons states. We revealed that the existence of symmetric, regular gap solitons in a 12-fold quasicrystal is limited by the geometrical size of the hexagon that forms the core of the dodecahedral cell, which is the building block of the quasicrystal. PMID- 12636842 TI - Interaction of a soliton with a localized gain in a fiber Bragg grating. AB - A model of a lossy nonlinear fiber grating with a "hot spot," which combines a local gain and an attractive perturbation of the refractive index, is introduced. A family of exact solutions for pinned solitons is found in the absence of loss and gain. In the presence of the loss and localized gain, an instability threshold of the zero solution is found. If the loss and gain are small, it is predicted what soliton is selected by the energy-balance condition. Direct simulations demonstrate that only one pinned soliton is stable in the conservative model, and it is a semiattractor: solitons with a larger energy relax to it via emission of radiation, while those with a smaller energy decay. The same is found for solitons trapped by a pair of repulsive inhomogeneities. In the model with the loss and gain, stable pinned pulses demonstrate persistent internal vibrations and emission of radiation. If these solitons are nearly stationary, the prediction based on the energy balance underestimates the necessary gain by 10-15% (due to radiation loss). If the loss and gain are larger, the intrinsic vibrations of the pinned soliton become chaotic. The local gain alone, without the attractive perturbation of the local refractive index, cannot maintain a stable pinned soliton. For collisions of moving solitons with the "hot spot," passage and capture regimes are identified, the capture actually implying splitting of the soliton. PMID- 12636843 TI - Soliton dynamics in a random Toda chain. AB - This paper addresses the soliton dynamics in a Toda lattice with a randomly distributed chain of masses. Applying the inverse scattering transform, we derive effective equations for the decay of the soliton amplitude that take into account radiative losses. It is shown that the soliton energy decays as approximately N( 3/2) for small-amplitude solitons and approximately exp(-N) for large-amplitude solitons. The decay rate does not depend on the incoming energy for large amplitude soliton. An important feature is the generation of a soliton gas consisting of a large collection of small solitons (a number of the order of epsilon (-2) of solitons with momenta of the order of epsilon (2), where epsilon is the strength of fluctuations). The soliton gas plays an important role in that the changes in the conservation equations cannot be correctly understood if the soliton production is neglected. The role of the correlation length of fluctuations on the soliton decay is discussed. It is shown that in the presence of long-range correlation the Toda soliton is not backscattered, but progressively converted into forward-going radiation. The analytical predictions are confirmed by full numerical simulations. PMID- 12636844 TI - Duality relation for the Maxwell system. AB - This paper is intended to establish a link between the vector Maxwell system for three-dimensional (3D) and 2D finite photonic crystals in the low-frequency limit. For this, we generalize the classical results of Keller and Dykhne (chessboard problem) to periodic media described by piecewise continuous permittivity profiles: our theorem enlights the result of Mendelson (polycrystalline and multiphase media) in the framework of homogenization theory of elliptic operators. In fine, we give illustrative examples by using both integral equation and variational approaches via the so-called method of fictitious charges and finite-element method. PMID- 12636845 TI - (2+1)-dimensional soliton formation in photorefractive Bi12SiO20 crystals. AB - (2+1)-dimensional spatial solitons in Bi12SiO20 (BSO) photorefractive crystals with large optical activity are experimentally demonstrated. The soliton formation when a Gaussian beam is injected at the input has been previously analyzed numerically and then experimentally investigated. We demonstrate analytically, numerically, and experimentally that by applying static electric biases of high values, the polarization rotation accelerates: this acceleration prevents the beam from broadening if the polarization rotation period becomes shorter than the diffraction length. Contemporary to this nonlinear optical activity, an induced birefringence affects the beam polarization state. Analysis of the polarization dynamics shows that the polarization changes nonuniformly across the beam (with a field dependent speed) until about 30-35 kV/cm; above this limit, the whole beam has just one polarization state. Representation on the Poincare sphere of the polarization dynamics reveals the existence of a stable polarization trajectory closed around a polarization attractor that depends on the linear optical activity and on the photorefractive nonlinearity. The experimental soliton is well described by the analytical solutions already obtained [Fazio et al., Phys. Rev. E 66, 016605 (2002)]. PMID- 12636846 TI - Degeneracy analysis for a supercell of a photonic crystal and its application to the creation of band gaps. AB - A method is introduced to analyze the degeneracy properties of the band structure of a photonic crystal by making use of supercells. The band structure associated with a supercell of a photonic crystal has degeneracies at the edge of the Brillouin zone if the photonic crystal has some kind of point group symmetry. The E-polarization and H-polarization cases have the same degeneracies for a two dimensional (2D) photonic crystal. Two theorems on degeneracies in the band structure associated with the supercell are given and proved. These degeneracies can be lifted to create photonic band gaps by changing the translation group symmetry of the photonic crystal (the point group symmetry of the photonic crystal may remain unchanged), which consequently changes the transform matrix between the supercell and the smallest unit cell. The existence of photonic band gaps for many known 2D photonic crystals is explained through the degeneracy analysis. Some structures with large band gaps are also found through the present degeneracy analysis. PMID- 12636847 TI - Universality of electromagnetic-field correlations within homogeneous and isotropic sources. AB - We investigate the structure of second-order correlations in electromagnetic fields produced by statistically stationary, homogeneous, and isotropic current distributions. We show that the coherence properties of such fields within a low loss or nondissipative medium do not depend on the source characteristics, but are solely determined by the propagation properties, and that the degree of coherence of the field is given by the sinc law. Our analysis reproduces the known results for blackbody fields, but it applies to a wider class of sources, not necessarily in thermal equilibrium. We discuss the physics behind the universal behavior of the correlations by comparing the results with those obtained by an electromagnetic plane-wave model. PMID- 12636848 TI - Numerical simulation of natural convection in a concentric annulus between a square outer cylinder and a circular inner cylinder using the Taylor-series expansion and least-squares-based lattice Boltzmann method. AB - In this paper, natural convective heat transfer in a horizontal concentric annulus between a square outer cylinder and a heated circular inner cylinder is numerically studied using the Taylor-series-expansion and least-squares-based lattice Boltzmann method (TLLBM). The TLLBM is used to extend the current thermal model to more practical applications. Since the TLLBM is basically a meshless approach and can be applied to any complex geometry, we can easily use it to solve the complex thermal problem accurately and effectively. The present method is validated by comparing its numerical results with available data in the literature, and very good agreement has been achieved. PMID- 12636849 TI - Nonequilibrium relaxation analysis of Kosterlitz-Thouless phase transition. AB - A simple and efficient numerical analysis is proposed for the Kosterlitz-Thouless (KT) phase transition. The nonequilibrium relaxation method is applied to it. The two-dimensional ferromagnetic XY models are investigated to show the efficiency. At the KT transition point as well as inside the KT phase, the nonequilibrium relaxation of magnetization from the all-aligned state shows an asymptotic power law decay, m(t) approximately t(-lambda(T)). Only outside the KT phase, an asymptotic single exponential decay is observed. Using a standard scaling form m(t)=tau(-lambda)(-)m(t/tau) in this regime, where tau is the relaxation time at each temperature, we find a simple and efficient numerical estimation of the KT transition point and dynamical exponent. This method can be applied to various kinds of models which show the KT-like behavior. PMID- 12636850 TI - Numerical approximation of breathers in lattices with nearest-neighbor interactions. AB - A numerical method is presented for accurately approximating time-periodic and spatially localized solutions known as (discrete) breathers, of the equations of motion describing lattices with one degree-of-freedom per lattice site and nearest-neighbor interactions. Our method is an extension of an approximation suggested by Tsironis [J. Phys. A 35, 951 (2002)], gives more accurate results than the rotating wave approximation and is more universally applicable for these lattices. As an illustration, the method is applied here to several one- and two dimensional lattices. PMID- 12636851 TI - Vlasov model using kinetic phase point trajectories. AB - A method of solution of the collisionless Vlasov equation by following fixed collisionless phase point ("particle") trajectories (characteristics) in phase space is presented. It solves the coupled Vlasov Maxwell system self-consistently and employs the Leapfrog-Trapezoidal scheme to solve for the characteristics explicitly. It then uses the bilinear finite element interpolation scheme in phase space and maps vital instantaneous phase point information (distribution function) to a fixed background phase space mesh while retaining it at the phase point. The scheme is an enhanced second order one in time and fourth order in space. The code is then used to model a thermal plasma as well as two stream instability using mobile electrons and fixed background ions: the scheme being a momentum conserving one by construction allows energy conservation without assignment of particle shape functions; Langmuir waves are obtained with very good agreement with the Bohm-Gross dispersion relation; the two stream results do not show any numerically induced oscillations attributed to the initial well ordered velocity distributions. Retention of the characteristics also minimized diffusion. Extensive numerical stability analysis deriving Courant condition for the scheme as well as behavior of computational modes are done in Appendix A, as well as estimating the impact of numerical diffusion in Appendix B. Two to five dimensional versions in phase space exist. PMID- 12636852 TI - Smoothed dissipative particle dynamics. AB - We present a fluid particle model that is both a thermodynamically consistent version of smoothed particle hydrodynamics (SPH) and a version of dissipative particle dynamics (DPD), capturing the best of both methods. The model is a discrete version of Navier-Stokes equations, like SPH, and includes thermal fluctuations, like DPD. This model solves some problems with the physical interpretation of the original DPD model. PMID- 12636853 TI - Convergence of reinforcement learning algorithms and acceleration of learning. AB - The techniques of reinforcement learning have been gaining increasing popularity recently. However, the question of their convergence rate is still open. We consider the problem of choosing the learning steps alpha(n), and their relation with discount gamma and exploration degree epsilon. Appropriate choices of these parameters may drastically influence the convergence rate of the techniques. From analytical examples, we conjecture optimal values of alpha(n) and then use numerical examples to verify our conjectures. PMID- 12636854 TI - Simulation of wave packet tunneling of interacting identical particles. AB - We demonstrate a different method of simulation of nonstationary quantum processes, considering the tunneling of two interacting identical particles, represented by wave packets. The used method of quantum molecular dynamics (WMD) is based on the Wigner representation of quantum mechanics. In the context of this method ensembles of classical trajectories are used to solve quantum Wigner Liouville equation. These classical trajectories obey Hamiltonian-like equations, where the effective potential consists of the usual classical term and the quantum term, which depends on the Wigner function and its derivatives. The quantum term is calculated using local distribution of trajectories in phase space, therefore, classical trajectories are not independent, contrary to classical molecular dynamics. The developed WMD method takes into account the influence of exchange and interaction between particles. The role of direct and exchange interactions in tunneling is analyzed. The tunneling times for interacting particles are calculated. PMID- 12636855 TI - Bilinear diffusion quantum Monte Carlo methods. AB - The standard method of quantum Monte Carlo for the solution of the Schrodinger equation in configuration space can be described quite generally as devising a random walk that generates-at least asymptotically-populations of random walkers whose probability density is proportional to the wave function of the system being studied. While, in principle, the energy eigenvalue of the Hamiltonian can be calculated with high accuracy, estimators of operators that do not commute the Hamiltonian cannot. Bilinear quantum Monte Carlo (BQMC) is an alternative in which the square of the wave function is sampled in a somewhat indirect way. More specifically, one uses a pair of walkers at positions x and y and introduces stochastic dynamics to sample phi(i)(x)t(x,y)phi(j)(y), where phi(i)(x) and phi(j)(y) are eigenfunctions of (possibly different) Hamiltonians, and t(x,y) is a kernel that correlates positions x and y. Using different Hamiltonians permits the accurate computation of small energy differences. We review the conceptual basis of BQMC, discuss qualitatively and analytically the problem of the fluctuations in the branching, and present partial solutions to that problem. Finally we exhibit numerical results for some model systems including harmonic oscillators and the hydrogen and helium atoms. Further research will be necessary to make this a practical and generally applicable scheme. PMID- 12636856 TI - Computational complexity arising from degree correlations in networks. AB - We apply a Bethe-Peierls approach to statistical-mechanics models defined on random networks of arbitrary degree distribution and arbitrary correlations between the degrees of neighboring vertices. Using the nondeterministic polynomial time hard optimization problem of finding minimal vertex covers on these graphs, we show that such correlations may lead to a qualitatively different solution structure as compared to uncorrelated networks. This results in a higher complexity of the network in a computational sense: Simple heuristic algorithms fail to find a minimal vertex cover in the highly correlated case, whereas uncorrelated networks seem to be simple from the point of view of combinatorial optimization. PMID- 12636858 TI - Morphologies resulting from the directional propagation of fractures. AB - When growing in a stress gradient, cracks have a directional growth. We investigate here this type of instability in the case of a colloidal gel deposited on a substrate and left to dry. The use of various materials reveals the existence of two distinct types of dynamics. When the crack nucleation is easy a well known situation is reached: an array of periodic fractures forms, which grow parallel to each other and move quasistatically with the stressed region. In contrast, in materials where the crack nucleation is difficult, a subcritical process is observed with the retarded formation of isolated cracks which move faster and which display an arch shaped trajectory. This type of process appears to be generic in all cases where there is delayed nucleation. This is confirmed by experiments on the directional propagation of cracks in thermally stressed glass. PMID- 12636857 TI - Statistics of the critical percolation backbone with spatial long-range correlations. AB - We study the statistics of the backbone cluster between two sites separated by distance r in two-dimensional percolation networks subjected to spatial long range correlations. We find that the distribution of backbone mass follows the scaling ansatz, P(M(B)) approximately M(-(alpha+1))(B)f(M(B)/M(0)), where f(x)=(alpha+etax(eta))exp(-x(eta)) is a cutoff function and M0 and eta are cutoff parameters. Our results from extensive computational simulations indicate that this scaling form is applicable to both correlated and uncorrelated cases. We show that the exponent alpha can be directly related to the fractal dimension of the backbone d(B), and should therefore depend on the imposed degree of long range correlations. PMID- 12636859 TI - Equivalence of ensembles in creation-annihilation nonequilibrium models. AB - We consider a class of stochastic models on a lattice in which particles are created and annihilated with given rates. The class include very distinct models such as the contact process and the stochastic Ising model. We show how to set up stochastic models with the dynamics that conserve the number of particles that are equivalent, in the thermodynamic limit, to the creation-annihilation models. We also obtain a formula that allows the calculation of the rates with respect to the ensemble of constant particle number. PMID- 12636860 TI - Noise-induced synchronization in realistic models. AB - Noise-induced synchronization is studied numerically in two realistic models--the Pikovsky-Rabinovich circuit model and the Hindmarsh-Rose neuron model. A different feature, single-variable-Jacobian matrix, is found in these two models and conditions are found by which two noncoupled identical systems can been synchronized by forcing with a common Gaussian noise term. PMID- 12636861 TI - Saddle-node ghost-induced low-frequency fluctuations in an external-cavity laser diode. AB - We investigate numerically the low-frequency fluctuation regime in a laser diode subject to optical feedback. We demonstrate that a saddle-node ghost can induce this regime. PMID- 12636862 TI - Breaking time for the quantum chaotic attractor. AB - A model of a quantum dissipative system is considered in the regime when the classical limit corresponds to a chaotic attractor, and the breaking time tau(Planck) of the classical-quantum correspondence is obtained. The model describes a periodically kicked harmonic oscillator (or a particle in a constant magnetic field) with a dissipation. Another analog of this problem is the dissipative kicked Harper model. It is shown that in the limit of the so-called dying attractor, the breaking time tau(Planck) can be arbitrarily large. PMID- 12636863 TI - Synchronization-based approach for estimating all model parameters of chaotic systems. AB - The problem of dynamic estimation of all parameters of a model representing chaotic and hyperchaotic systems using information from a scalar measured output is solved. The variational calculus based method is robust in the presence of noise, enables online estimation of the parameters and is also able to rapidly track changes in operating parameters of the experimental system. The method is demonstrated using the Lorenz, Rossler chaos, and hyperchaos models. Its possible application in decoding communications using chaos is discussed. PMID- 12636864 TI - Correlation between Kolmogorov-Sinai entropy and self-diffusion coefficient in simple fluids. AB - The relationship between the Kolmogorov-Sinai entropy, h(KS) and the self diffusion coefficient D is studied for two classical simple fluid systems with purely repulsive potentials (one system with a Wayne-Chandler-Anderson potential and the other with a hard-sphere potential). Numerical simulation data for h(KS) and D, normalized by the average collision frequency nu and the diameter of the particle sigma as natural units of time and distance, reveal that, in the region spanning from normal liquid up to near solidification (0.50< or =rho< or =0.93), the Kolmogorov-Sinai entropy has a power law dependeney on the self-diffusion coefficient of the form h(KS)/nu proportional, variant (D/sigma(2)nu)(eta), in which eta is independent of density and temperature. PMID- 12636865 TI - Direct measurements of the energy of intense vorticity filaments in turbulence. AB - The global instantaneous power injected in a turbulent shear flow is investigated. A conditional averaging technique reveals the complete energetic transfer scenario of the intense vorticity filaments, which were visualized by Douady, Couder, and Brachet [Phys. Rev. Lett. 67, 983 (1991)]. It is found that during their existence filaments store a significant part of the turbulent kinetic energy. This energy is dissipated during the destruction phase of filaments. The total energy transferred to small scales due to filaments is quantified on the basis of a global flow visualization. PMID- 12636866 TI - Oscillatory mean-field dynamos with a spherically symmetric, isotropic helical turbulence parameter alpha. AB - Until recently, the existence of oscillatory mean-field dynamos of the alpha(2) type with spherically symmetric and isotropic turbulence parameter alpha was an open question. We find such dynamos by means of an evolutionary strategy, and we illustrate the spectral properties of the corresponding dynamo operators. PMID- 12636867 TI - Oscillatory Rayleigh-Marangoni convection in a layer heated from above: numerical simulations with an undeformable free surface. AB - Joint action of buoyancy and thermocapillary forces can destabilize the motionless state in a liquid layer heated from above due to the coupling of internal and surface waves. The nonlinear evolution of this oscillatory instability is studied using three-dimensional direct numerical simulations with a pseudospectral Fourier-Chebyshev code. Alternating rolls and standing, oscillating squares are observed as final convective patterns. The flow is strongly localized near the free surface. Buoyancy plays a negligible role in kinetic energy production. PMID- 12636868 TI - Octupolar perturbation of a single ion in a Penning trap. AB - We investigate the classical dynamics of a single ion trapped in a Penning trap perturbed by a octupolar electrostatic perturbation that introduces nonlinearities in the motion. We show that the dynamics is controlled by a single external parameter that combines the influence of the electric and magnetic fields. Through the variation of this parameter, we explore the evolution of the phase space structure of the system by the numerical continuation of the families of periodic orbits. PMID- 12636869 TI - Model of nonlinear kinetic Alfven waves with dissipation and acceleration of energetic electrons. AB - The acceleration of energetic electrons has long been one of the most outstanding problems in astrophysics and space physics, and some recent observations from space satellites show that low-frequency electromagnetic fluctuations in the auroral ionosphere and magnetosphere can often be identified as the kinetic Alfven modes. A model of nonlinear kinetic Alfven waves is presented here, in which the effect of electron collisional dissipation has been taken into account. The result is a dissipative solitary kinetic Alfven wave (DSKAW), which can produce a local shocklike structure with a net electric potential drop and which can thereby accelerate efficiently the electrons to the order of the local Alfven velocity. Since Alfvenic fluctuation is the most common electromagnetic activity in extensive cosmic plasma environments, the present result suggests that the DSKAW could play an important role in the acceleration and energization of cosmic plasmas. PMID- 12636870 TI - Rice's ansatz for overdamped phi4 kinks at finite temperature. AB - The dynamics of a kink driven by noise is analyzed using the two collective variables of the Rice ansatz: position and width. Starting from a stochastic partial differential equation, with the phi(4) potential in the overdamped limit, the pair of stochastic differential equations for the collective variables are derived without approximation other than the ansatz itself. From the steady state probability density of the kink width, the diffusivity of a kink is calculated. PMID- 12636871 TI - Josephson junction coupled to a transmission line: a comparison of different approaches. AB - The case of a Josephson junction loaded by a transmission line is reexamined, according to the Green's function method, in order to compare the results with those that we previously obtained, analytically and numerically, following a different procedure. PMID- 12636872 TI - Effects of anti-proteinuric therapy with angiotensin-converting-enzyme inhibition on renal protein catabolism in the adriamycin-induced nephrotic rat. AB - A direct consequence of glomerular protein leakage is an increased exposure of proximal tubular cells to proteins. The aim of the present study was to examine whether chronic proteinuria affects the tubular handling of proteins and whether anti-proteinuric therapy by angiotensin-converting-enzyme (ACE) inhibition restores this tubular function. Renal uptake and catabolic rate of the low molecular-weight protein (LMWP) myoglobin was determined in anaesthetized control and adriamycin-induced nephrotic rats by external counting after radiolabelling. Proteinuria correlated with the uptake as well as the catabolism of myoglobin. The higher the proteinuria, the lower was the renal uptake of myoglobin (r =0.72, P =0.002). Also, the catabolic rate of myoglobin (r =0.80, P =0.0002) was lower with increasing severity of proteinuria. During treatment with the ACE inhibitor lisinopril, proteinuria was lowered by 79+/-9% (mean+/-S.E.M.). Renal uptake and catabolic rate of the LMWP were not restored by ACE inhibition. The catabolic rate of myoglobin was even decreased further with 48+/-5% compared with pretreatment levels. In summary, adriamycin-induced proteinuria is associated with a lower uptake and a lower catabolic rate of LMWP in the proximal tubule. ACE inhibition lowers proteinuria, but does not restore the affected LMWP uptake and rate of catabolism. The rate of LMWP catabolism is even decreased further. In contrast, the urinary excretion of N -acetyl glucosaminidase, the tubular marker of toxicity, was effectively returned to normal levels during ACE inhibition. Taken together, the data suggest that proteinuria is toxic for the proximal tubular cells and that ACE inhibition protects the remaining functional tubular cells directly against destruction through decreasing hypercatabolism. PMID- 12636874 TI - Animated documentation of the filaria dance sign (FDS) in bancroftian filariasis. AB - BACKGROUND: Ultrasonography is able to detect adult Wuchereria bancrofti worms in scrotal lymphatic vessels of infected men on account of the characteristic pattern of adult worm movements, known as the filarial dance sign. Furthermore, the technique is able to delineate associated pathology, such as hydrocoele and lymphoedema, which can be diagnosed in early stages. Ultrasonography is also useful in the assessment of macrofilaricidal effects of antifilarial medication.The purpose of this study was to evaluate the usefulness of scrotal ultrasonography, in combination with a new method of digital documentation, in men infected with Wuchereria bancrofti. METHODS: Ultrasonography of the scrotal areas was carried out in 33 male patients from an endemic area in Ghana using a hand-carried ultrasound system and a linear array transducer at 7.5 MHz. Wuchereria bancrofti infection was also assessed by quantification of night blood microfilaraemia and semi-quantitative detection of circulating filarial antigen. Ultrasound findings were documented by print outs and by Digital Video sequences directly exported from the ultrasound machine which were edited in Final Cut Pro 3ledR; and exported, using QuickTimecircledR; Pro, as MPEG-1 video. RESULTS: Worm nests, i.e. dilated lymphatic vessels with the characteristic movement patterns of worms, were found in all patients, and typical examples of larger as well as smaller nests are presented through MPEG-1 video in b- and m-modes as well as Colour Doppler and Pulse Wave Doppler images. CONCLUSION: In this study, the filarial dance sign is being made available on the Internet to readers through MPEG-1 video. This method allows for demonstration of movement patterns rather than static images. In addition, the pathologic ultrasonographic signs of filariasis can be rapidly relayed over great distances and may be helpful to other investigators or clinicians in the diagnosis of patients infected with Wuchereria bancrofti. PMID- 12636875 TI - Observations on the swarming and mating behaviour of Anopheles funestus from southern Mozambique. AB - BACKGROUND: Control of malaria by the release of genetically modified mosquitoes refractory to transmission is now becoming a possibility. In many areas of Africa, Anopheles gambiae is found together with an equally important vector, An. funestus. Given their sympatry and the likelihood of a similar mating period some aspects of the mating behaviour of An. gambiae s.l. and An. funestus are likely to differ. We therefore attempted to characterise the swarming behaviour of An. funestus and to determine if any aspects of the observed behaviour differed from that recorded for the M form of An. gambiae from Sao Tome. METHODS: In March - May 2002 the swarming, mating, house exiting and resting behaviour of Anopheles funestus was studied by direct observation in Mozambique. Swarming males and insects in copula were collected by sweep net. Wing lengths of males collected resting, exiting houses, swarming and mating were measured and the wingbeat frequency distribution of individual insects, in free flight confined inside netting covered paper cups, was also determined. RESULTS: Mono-specific swarms occurred at sunset in relatively open areas close to houses used for resting. Mating pairs were seen 11 +/- 3.7 min after the start of swarming. The number of total pairs observed being inversely proportional to the time difference between the start of swarming and the first pairing. The great majority of females mated before feeding. Male or female size did not appear to affect mating success or other behaviours. During the study, ambient temperatures decreased and female, but not male, wing size increased. At 516 Hz, the flight tone of female An. funestus was similar to the 497 Hz of the local An. gambiae. Males dispersed if light or dark artificial horizontal markers were placed underneath naturally occurring swarms. CONCLUSION: Differential response to markers would be sufficient for swarming in An. funestus and An. gambiae s.l. to occur in distinct sites. PMID- 12636873 TI - What is the origin of pancreatic adenocarcinoma? AB - The concept of pancreatic cancer origin is controversial. Acinar, ductal or islet cells have been hypothesized as the cell of origin. The pros and cons of each of these hypotheses are discussed. Based on the world literature and recent observations, pancreatic cells seem to have potential for phenotypical transdifferentiation, i.e ductal-islet, ductal-acinar, acinar-ductal, acinar islet, islet-acinar and islet-ductal cells. Although the possibility is discussed that cancer may arise from either islet, ductal or acinar cells, the circumstances favoring the islet cells as the tumor cell origin include their greater transdifferentiation potency into both pancreatic and extrapancreatic cells, the presence of a variety of carcinogen-metabolizing enzymes, some of which are present exclusively in islet cells and the growth factor-rich environment of islets. PMID- 12636876 TI - A comparison between the effort-reward imbalance and demand control models. AB - BACKGROUND: To compare the predictive validity of the demand/control and reward/imbalance models, alone and in combination with each other, for self reported health status and the self-reported presence of any chronic disease condition. METHODS: Self-reports for psychosocial work conditions were obtained in a sample of sawmill workers using the demand/control and effort/reward imbalance models. The relative predictive validity of task-level control was compared with effort/reward imbalance. As well, the predictive validity of a model developed by combining task-level control with effort/reward imbalance was determined. Logistic regression was utilized for all models. RESULTS: The demand/control and effort/reward imbalance models independently predicted poor self-reported health status. The effort-reward imbalance model predicted the presence of a chronic disease while the demand/control model did not. A model combining effort-reward imbalance and task-level control was a better predictor of self-reported health status and any chronic condition than either model alone. Effort reward imbalance modeled with intrinsic effort had marginally better predictive validity than when modeled with extrinsic effort only. CONCLUSIONS: Future work should explore the combined effects of these two models of psychosocial stress at work on health more thoroughly. PMID- 12636877 TI - Time dependent ethnic convergence in colorectal cancer survival in Hawaii. AB - BACKGROUND: Although colorectal cancer death rates have been declining, this trend is not consistent across all ethnic groups. Biological, environmental, behavioral and socioeconomic explanations exist, but the reason for this discrepancy remains inconclusive. We examined the hypothesis that improved cancer screening across all ethnic groups will reduce ethnic differences in colorectal cancer survival. METHODS: Through the Hawaii Tumor Registry 16,424 patients diagnosed with colorectal cancer were identified during the years 1960-2000. Cox regression analyses were performed for each of three cohorts stratified by ethnicity (Caucasian, Japanese, Hawaiian, Filipino, and Chinese). The models included stage of diagnosis, year of diagnosis, age, and sex as predictors of survival. RESULTS: Mortality rates improved significantly for all ethnic groups. Moreover, with the exception of Hawaiians, rates for all ethnic groups converged over time. Persistently lower survival for Hawaiians appeared linked with more cancer treatment. CONCLUSION: Ethnic disparities in colorectal cancer mortality rates appear primarily the result of differential utilization of health care. If modern screening procedures can be provided equally to all ethnic groups, ethnic outcome differences can be virtually eliminated. PMID- 12636878 TI - Bone pain associated with once-per-cycle pegfilgrastim is similar to daily filgrastim in patients with breast cancer. AB - Bone pain is a common side effect of treatment with filgrastim. Pegfilgrastim is a pegylated long-acting analogue of filgrastim that is administered once per chemotherapy cycle. The profile of prospectively defined, patient-reported bone pain judged by the investigators as related to study drug was analyzed retrospectively for each drug using data from two comparable phase III trials. These multicenter, randomized, double-blind, noninferiority trials compared once per-cycle pegfilgrastim (6 mg, study 1 or 100 microg/kg, study 2) to daily filgrastim 5 microg/kg in patients with stage II-IV breast cancer undergoing multiple cycles of myelosuppressive chemotherapy (doxorubicin/docetaxel). Subcutaneous once-per-cycle pegfilgrastim 6-mg and 100-microg/kg doses were administered to 76 and 150 patients, respectively; subcutaneous daily filgrastim 5 microg/kg was administered to a total of 227 patients. Because bone pain in study 1 was higher (P = 0.044) in every cycle compared with study 2, all analyses were performed separately for each study. No statistically significant differences in incidence, severity, or duration were observed between patients receiving either once-per-cycle pegfilgrastim or daily filgrastim in either study. Bone pain incidence and severity were significantly greater (P < 0.001) in cycle 1 of both studies compared with later cycles. Among patients with bone pain, a trend towards earlier onset with pegfilgrastim was observed but was not associated with increased bone pain severity or duration. In patients who received a fixed 6-mg dose of pegfilgrastim, the overall bone pain incidence was similar when analyzed by body weight (< 60 kg, 60-100 kg, > 100 kg). No patients were withdrawn from either study for bone pain. PMID- 12636879 TI - Chemotherapy for breast cancer: what's new? What's useful? PMID- 12636883 TI - Prognostic and predictive indicators in operable breast cancer. AB - Because of its biological heterogeneity and wide spectrum of responsiveness to different treatments, breast cancer is a complex disease of difficult clinical management. Over the past several years, knowledge of the molecular mechanisms regulating normal and aberrant cell growth leading to cancer has been enhanced. These advances have enabled the identification of an increasing number of surrogate biomarkers, which have been correlated with prognosis or used as predictors of response to specific treatments. Axillary nodal status, age, tumor size, pathologic grade, and hormone receptor status are the established prognostic and/or predictive factors for selection of adjuvant treatments. The role of new biomarkers, such as p53, HER2/neu, angiogenesis, and the proliferation index value, is promising; however, the clinical value of their determination must be provided by prospective clinical studies. PMID- 12636884 TI - Vinorelbine as a 96-hour continuous infusion in heavily pretreated patients with metastatic breast cancer: a cooperative study by the GEICAM group. AB - The efficacy of vinorelbine given as a continuous infusion was evaluated in 47 patients with breast cancer who had received previous treatment with first-line, second-line, and third-line chemotherapy including taxanes and/or anthracyclines. For inclusion into the study, patients were required to have histology-proven bi dimensionally measurable disease. The treatment schedule was a bolus injection of vinorelbine 8 mg/m(2) administered over 5-10 minutes on day 1 followed by vinorelbine 8 mg/m(2) continuous infusion on days 1-4, every 21 days for 6 cycles. On an intent-to-treat basis, a 2% complete response rate and a 17% partial response rate were observed. The median time to progression was 2.4 months (95% CI, 1.83-2.97). Median survival was 7.73 months (95% CI, 4.48-10.98; range, 0.33-55.0 months). Major toxicities included febrile neutropenia in 40 cycles (24%) affecting 24 patients (51%) and 1 toxic death. Other nonhematologic toxicities included stomatitis (13%) and asthenia (13%). We conclude that this treatment shows considerable therapeutic activity, albeit at considerable toxicity costs, even in patients who have had multiple lines of prior chemotherapy. However, the results do not indicate clear advantages compared to the conventional weekly scheme of administration. PMID- 12636885 TI - Inhibiting Ras signaling in the therapy of breast cancer. AB - Ras is a small guanosine triphosphate-binding protein that plays an important role in signal transduction pathways that influence cellular proliferation, apoptosis, cytoskeletal organization, and other important biological processes. Prenylation of Ras proteins by the enzyme farnesyltransferase renders the protein hydrophobic, causing localization to the inner surface of the cell membrane, where it exerts its biological effects. Ras mutations that result in constitutive activation of the Ras pathway are common in certain human cancers, and transfection of cell lines with mutant Ras renders them tumorigenic. Farnesyltransferase inhibitors (FTIs) were initially developed to inhibit growth of cancers harboring Ras mutations, but preclinical data suggests that they also have antiproliferative effects in cell lines with wild-type Ras. Preclinical data suggest that FTIs have antiproliferative and antitumor effects in breast cancer cell lines, but the precise target(s) remain to be defined. One phase II trial has demonstrated that one orally administered FTI has significant antitumor activity in metastatic breast cancer. In addition, preclinical evidence suggests that FTIs may augment the activity of cytotoxic agents and hormonal therapy. Clinical trials are currently underway evaluating whether these agents have a useful role in the management of advanced breast cancer. PMID- 12636887 TI - E2100: a phase III trial of paclitaxel versus paclitaxel/bevacizumab for metastatic breast cancer. PMID- 12636888 TI - A study of computer-based task performance under thermal stress. AB - A visual-visual dual computer task was designed to test the effect of the thermal environment on dual task performance. The temperatures selected for testing were 20 and 35 degrees C Wet Bulb Globe Temperature (WBGT). 34 volunteers were randomly assigned to 1 of the 2 temperature conditions. Individual differences in single task performance were controlled by equating the baselines of single task performance. Once individual differences in single task capacity were controlled, statistically significant differences in performance were demonstrated. Mean accuracy was computed over a 1-hr testing period in each temperature condition. Participants' mean accuracy in the 35 degrees condition (38.18%) was substantially lower than in the 20 degrees condition (50.88%). PMID- 12636890 TI - Assessing the compatibility of work system factors through an integrative model: a case study. AB - Work Factor Compatibility (WFC) theory proposes that human performance is dependent on the interaction of various components (e.g., physical and mental task demands, physical environment, social environment) of the work system and integrates multiple human performance perspectives into a single mathematical model. Work Factor Analysis (WFA) is a comprehensive survey administered to employees to determine the WFC index. WFA also provides recommendations for targeting specific work system areas for improvement. Preliminary testing of the tool was conducted at a manufacturing operation of a Fortune 10 company. Results are discussed and recommendations for further study are made. PMID- 12636889 TI - Occupational exposure to polycyclic aromatic hydrocarbons during Diesel combustion. AB - Identification and determination of polycyclic aromatic hydrocarbons (PAHs) in Diesel exhaust in the working environment and assessment of workers' occupational exposure to these suspected human carcinogens were the aim of this experimental investigation. The range of exposure factors calculated on the basis of 9 individual PAH concentrations determined in personal air samples shows that time averaged concentration of these compounds did not exceed the Polish Maximum Admissible Concentration (MAC) value for PAHs, that is, 2 microg.sup-3;. The highest concentrations of PAHs were determined in the breathing zone of forklift operators. The maximum exposure factor was 0.427 microg.msup-3; (about 1/4 of MAC). PMID- 12636891 TI - Enhancement of drilling safety and quality using online sensors and artificial neural networks. AB - Cutting force sensors and neural networks have been used for the occupational safety of the drilling process. The drill conditions have been online classified into 3 categories: safe, caution, and danger. This approach can change the drill just before its failure. The inputs to neural networks include drill size, feed rate, spindle speed, and features that were extracted from drilling force measurements. The outputs indicate the safety states. This detection system can reach a success rate of over 95%. Furthermore, the one misclassification during online tests was a one-step ahead pre-alarm that is acceptable from the safety and quality viewpoint. The developed online detection system is very robust and can be used in very complex manufacturing environments. PMID- 12636893 TI - Health risks from exposure to metal-working fluids in machining and grinding operations. AB - Metal-working fluids (MWFs) are used in machining and grinding operations to cool the tool and work, reduce the friction between the tool and work, improve the surface integrity of the work piece, and increase tool life and productivity. Health problems have been reported among workers exposed to MWFs, including incidences of respiratory, digestive and skin cancers, and increased rates of cough and phlegm. This paper reviews and discusses issues concerning health risks from exposure to MWFs in machining and grinding operations, the various factors that influence the degree of exposure, and control methods to reduce exposure to metal-working fluids. PMID- 12636892 TI - Ergonomic study on the manual component insertion lines for occupational health and safety improvements. AB - The study aimed at reducing the occupational health and safety problems faced by the manual component insertion operators. Subjective and objective assessments, and direct observations were made in the printed circuit assembly factory. Simple and low-cost ergonomic interventions were implemented, that is, repairing chairs, reducing high workloads, assigning operators to a maximum of 2 workstations, confining machines that emitted bad smell and much noise, and providing finger work aids. The results of the interventions were reductions in operators' work discomforts, that is, chair discomfort (by 90%), high work stress, and discomfort due to profound change in their workstations. Their health hazards were also eliminated, that is, inhalation of toxic fumes, exposure to too much noise, and pain due to pressing sharp components. PMID- 12636894 TI - Asbestos-related health risks in Estonia. AB - Asbestos has been declared a proven human carcinogen by the U.S. Environmental Protection Agency and by the International Agency for Research on Cancer of the World Health Organization. Since 1983, the European Union has enforced health protection directions in this field, which have been the basis for legal acts of Member States. The Estonian Republic is a country where asbestos and asbestos containing materials have been used in quite large quantities, especially during the Soviet period. Today the use and sale of asbestos minerals and asbestos containing products are prohibited in Estonia. Unfortunately there has been no substantial overview of the health status of workers exposed to asbestos. The objective of this study was to evaluate the extent of health risks of workers exposed to asbestos in Estonia. PMID- 12636895 TI - Work-related research, education, and training in developing countries. AB - Work-related research, education, and training (WRET) have not been widely recognised in many developing countries (DCs) as the most important factor for sustainable workplace improvement. There are many reasons why WRET is still neglected or remains unrecognised. Empirical research, advanced studies, and training abroad do not seem to be cost-effective for many people living in DCs because of enormous obstacles. Therefore, it is not easy to demonstrate that WRET result in workplace improvement in diverse situations in each DC. Taking into consideration poor health and safety in various workplaces, this paper aims to stimulate critical opinions and discussions on WRET, which are yet to be given high priority in the national agenda to ensure industrial production and social progress. PMID- 12636896 TI - [Non invasive ventilation for acute exacerbation of chronic obstructive pulmonary disease: a meta-analysis]. AB - BACKGROUND AND OBJECTIVES: The purpose of this study was: 1) To determine the effect of non invasive ventilation (NIV) in chronic obstructive pulmonary disease (COPD) patients with acute respiratory failure, and 2) To find out whether there are differences in the results according to the site where ventilation in applied. PATIENTS AND METHOD: Meta-analysis of randomized controlled trials (RCTs) published included studies in MEDLINE, EMBASE, Cochrane Controled Trials Register and Teseo. Study selection included RCTs in COPD patients with acute respiratory failure with PaCO2 > 45 mmHg or pH < 7.35, with underwent NIV versus standard therapy. Mortality, rate of tracheal intubation and length of stay were analized. Individual studies were rated as to their validity and quality by two investigators using the Jadad score. A different meta-analysis was made for intensive-care unit (ICU) RCTs and wards RCTs. RESULTS: Five ICU RCTs and four ward RCTs were included. There was a reduction in hospital mortality (ICU, OR = 0.35; 95% confidence interval [CI], 0.15 to 0.83; n = 166; p = 0.02; ward, OR = 0.39; 95% CI, 0.20 to 0.76; n = 296; p = 0.005) and in the need for intubation (ICU, OR = 0.18; 95% CI, 0.10 to 0.35; n= 219; p < 0,00001; ward, OR = 0.43; IC 95%, 0.24-0.78; n = 270; p = 0.006). Length of stay was lower in the NIV group upon ICU trials (WMD, 3.7 days; 95% CI, 5.46 to 1.93 days). Only one ward RCT showed significant differences in the length of stay. Published data did not allow a meta-analysis for length of stay. CONCLUSIONS: NIV decreases mortality and the need for intubation in COPD patients with acute respiratory failure both in ICU and ward. In ICU non-invasive ventilation decreases the length of stay too. PMID- 12636897 TI - [Screening ability of a questionnaire designed for the detection of diabetes mellitus in hospital outpatients clinic]. AB - BACKGROUND AND OBJECTIVE: The socio-economical burden of type 2 diabetes mellitus (DM2) recommends its screening when the disease is suspected. In the present study we aimed to evaluate: the results of the oral glucose tolerance test (OGTT) normally indicated in hospital outpatient clinics when diabetes is suspected, and the efficacy of the American Diabetes Association (ADA) diabetes risk questionnaire to detect glucose tolerance abnormalities. PATIENTS AND METHOD: All subjects with an indication of OGTT were included consecutively. Individuals filled up a questionnaire including the ADA items. The sensitivity and specificity of the questionnaire were calculated. RESULTS: Two hundred and twenty nine subjects were studied (62.9% women): 44.1% had normal glucose tolerance (NGT) with normal fasting glucose. Moreover, 6.5%, 25.8% and 23.6% showed an impaired fasting glucose, impaired glucose tolerance and diabetes, respectively. According to the questionnaire, 45.8% of subjects undergoing the OGTT had a positive risk of diabetes. The questionnaire revealed a sensitivity = 72.2%, specificity = 60.6% and a positive predictive value = 37.1%. Age was the only variable that increased the risk of DM (odds ratio = 3.48, p = 0.0001). CONCLUSIONS: The important proportion of patients with NGT found in our study suggests the need to improve the indication of a diagnosis test when diabetes is suspected. One possibility would be to validate a questionnaire to detect the risk of diabetes in our own area. PMID- 12636899 TI - [Syphilis again?]. PMID- 12636898 TI - [Pneumococcal pneumonia in hospitalized patients. Therapeutic implications of resistances to penicillin and erythromycin]. AB - BACKGROUND AND OBJECTIVE: Streptococcus pneumoniae is the main etiological agent of community-acquired pneumonia. The aim of this work was to ascertain the resistance profiles of pneumococcus to penicillin and erythromycin and to analyse whether such profiles lead to different disease developments. PATIENTS AND METHOD: A retrospective analysis was carried out in 75 cases of pneumococcal pneumonia corresponding to hospitalized patients. Comorbidity factors were evaluated including their influence on the appearance of resistance. RESULTS: 67 patients (89.3%) presented comorbidity factors. 49.3% isolates displayed some type of resistance: 38.6% to penicillin, 36% to erythromycin and 13.3% to cefotaxime. No relationship was observed between the severity of the pneumonia and antibiotic resistance. Complications and mortality were not influenced by the susceptibility of pneumococcus to antibiotics. CONCLUSION: The increase in the resistance to antibiotics, especially erythromycin, makes betalactams the best choice for the treatment of pneumococcal pneumonia. PMID- 12636900 TI - [Community-acquired pneumococcal infection: approach to improve the election of antibiotic therapy]. PMID- 12636901 TI - [Hypertriglyceridemia as a cardiovascular risk factor. The end of a controversy?]. PMID- 12636902 TI - [Relapsing hypoglycemia in a 72-year-old male with diabetes mellitus]. PMID- 12636903 TI - [Subarachnoid hemorrhage and clopidogrel]. PMID- 12636905 TI - [Scedosporium prolificans: disseminated infection in immunocompromised patient]. PMID- 12636906 TI - [Inclusion body myositis and primary Sjogren syndrome]. PMID- 12636907 TI - [Health professionals and the consequences of a possible war in Iraq: open letter to the Spanish government]. PMID- 12636908 TI - RhoD, Src, and hDia2C in endosome motility. AB - The positioning of early endosomes is a dynamic process requiring coordinated activity of both actin and microtubules with their associated motors. A recent report from Marino Zerial and colleagues examining RhoD identifies mDia2C, a novel splice variant of mDia2, and Src as components of the regulatory machinery influencing actin-dependent endosome movement. PMID- 12636909 TI - Kar9 asymmetrical loading on spindle poles mediates proper spindle alignment in budding yeast. AB - In the February 21 issue of Cell, demonstrate that asymmetrical loading of Kar9 onto astral microtubules (MTs) emanating from the bud-ward-directed spindle pole ensures delivery of this spindle pole to the bud. Kar9 mediates alignment of the spindle with the cell polarity axis through a Myo2-dependent mechanism that reorients astral MTs toward the bud. PMID- 12636910 TI - Sex and the MHC. AB - Major histocompatibility complex class Ib molecules may play a surprising role in pheromone detection in mammals. PMID- 12636911 TI - Follow the leader. AB - Upon starvation, individual Dictyostelium amoebae chemotax toward aggregation centers in tightly packed streams in which cells are organized in head-to-tail chains. A recent report in Cell shows that this behavior requires localization of adenylyl cyclase and the production and secretion of the chemoattractant cAMP at the posterior of individual cells. These findings suggest a relay and communication system to regulate the long-range coordinated movement of cells. PMID- 12636912 TI - Visualization of the spatial and temporal dynamics of intracellular signaling. AB - In recent years, our ability to unravel the finer details of intracellular signaling has improved remarkably. Technological innovations resulting from the introduction of green fluorescent protein (GFP) have played a significant role in these advances. Fluorescent indicators allow us to visualize events within a cell in real-time and space. This review focuses on indicators that use GFP-based fluorescence resonance energy transfer (FRET) technologies and discusses how these methodologies have given insights into biological questions relating to the spatiotemporal patterns of signaling by cAMP, calcium, receptor tyrosine kinases, and other molecules. PMID- 12636913 TI - Drosophila gurken (TGFalpha) mRNA localizes as particles that move within the oocyte in two dynein-dependent steps. AB - In Drosophila oocytes, gurken mRNA localization orientates the TGF-alpha signal to establish the anteroposterior and dorsoventral axes. We have elucidated the path and mechanism of gurken mRNA localization by time-lapse cinematography of injected fluorescent transcripts in living oocytes. gurken RNA assembles into particles that move in two distinct steps, both requiring microtubules and cytoplasmic Dynein. gurken particles first move toward the anterior and then turn and move dorsally toward the oocyte nucleus. We present evidence suggesting that the two steps of gurken RNA transport occur on distinct arrays of microtubules. Such distinct microtubule networks could provide a general mechanism for one motor to transport different cargos to distinct subcellular destinations. PMID- 12636914 TI - The structure of the GGA1-GAT domain reveals the molecular basis for ARF binding and membrane association of GGAs. AB - The GGAs are a family of clathrin adaptor proteins involved in vesicular transport between the trans-Golgi network and endosomal system. Here we confirm reports that GGAs are targeted to the Golgi via interaction between the GGA-GAT domain and ARF-GTP, and we present the structure of the GAT domain of human GGA1, completing the structural description of the folded domains of GGA proteins. The GGA-GAT domain possesses an all alpha-helical fold with a "paper clip" topology comprising two independent subdomains. Structure-based mutagenesis demonstrates that ARF1-GTP binding by GGAs is exclusively governed by the N-terminal "hook" subdomain, and, using an in vitro recruitment assay, we show that ARF-GTP binding by this small structure is required and sufficient for Golgi targeting of GGAs. PMID- 12636915 TI - Centrosome separation and central spindle assembly act in redundant pathways that regulate microtubule density and trigger cleavage furrow formation. AB - The mitotic spindle provides the spatial cue that coordinates cytokinesis with nuclear division. However, the specific property of the mitotic spindle that mediates this spatial regulation remains obscure, in part because different aspects of the mitotic spindle appear to have furrow inducing activity in different systems. We show that in C. elegans embryos, although the central spindle is usually dispensable for furrow initiation, it becomes essential for furrow formation when the extent of centrosome separation during anaphase is reduced. Measurements of microtubule density demonstrate that furrow formation occurs in the vicinity of a local minimum of microtubule density. Reduction of the extent of spindle elongation or disruption of the central spindle causes delayed formation of the cleavage furrow. These data suggest that reduced microtubule density triggers cleavage furrow initiation and demonstrate that redundant mechanisms direct efficient formation of the cleavage furrow. PMID- 12636916 TI - Phosphorylation-dependent regulation of septin dynamics during the cell cycle. AB - Septins are GTPases involved in cytokinesis. In yeast, they form a ring at the cleavage site. Using FRAP, we show that septins are mobile within the ring at bud emergence and telophase and are immobile during S, G2, and M phases. Immobilization of the septins is dependent on both Cla4, a PAK-like kinase, and Gin4, a septin-dependent kinase that can phosphorylate the septin Shs1/Sep7. Induction of septin ring dynamics in telophase is triggered by the translocation of Rts1, a kinetochore-associated regulatory subunit of PP2A phosphatase, to the bud neck and correlates with Rts1-dependent dephosphorylation of Shs1. In rts1 Delta cells, the actomyosin ring contracts properly but cytokinesis fails. Together our results implicate septins in a late step of cytokinesis and indicate that proper regulation of septin dynamics, possibly through the control of their phosphorylation state, is required for the completion of cytokinesis. PMID- 12636917 TI - Cell cycle withdrawal, progression, and cell survival regulation by EGFR and its effectors in the differentiating Drosophila eye. AB - Receptor tyrosine kinases such as the EGF receptor transduce extracellular signals into multiple cellular responses. In the developing Drosophila eye, EGFR activity triggers cell differentiation. Here we focus on three additional cell autonomous aspects of EGFR function and their coordination with differentiation, namely, withdrawal from the cell cycle, mitosis, and cell survival. We find that, whereas differentiation requires intense signaling, dependent on multiple reinforcing ligands, lesser EGFR activity maintains cell cycle arrest, promotes mitosis, and protects against cell death. Each response requires the same Ras, Raf, MAPK, and Pnt signal transduction pathway. Mitotic and survival responses also involve Pnt-independent branches, perhaps explaining how survival and mitosis can occur independently. Our results suggest that, rather than triggering all or none responses, EGFR coordinates partially independent processes as the eye differentiates. PMID- 12636918 TI - Netrin-1/neogenin interaction stabilizes multipotent progenitor cap cells during mammary gland morphogenesis. AB - Netrin-1 and its receptors play an essential role patterning the nervous system by guiding neurons and axons to their targets. To explore whether netrin-1 organizes nonneural tissues, we examined its role in mammary gland morphogenesis. Netrin-1 is expressed in prelumenal cells, and its receptor neogenin is expressed in a complementary pattern in adjacent cap cells of terminal end buds (TEBs). We discovered that loss of either gene results in disorganized TEBs characterized by exaggerated subcapsular spaces, breaks in basal lamina, dissociated cap cells, and an increased influx of cap cells into the prelumenal compartment. Cell aggregation assays demonstrate that neogenin mediates netrin-1-dependent cell clustering. Thus, netrin-1 appears to act locally through neogenin to stabilize the multipotent progenitor (cap) cell layer during mammary gland development. Our results suggest that netrin-1 and its receptor neogenin provide an adhesive, rather than a guidance, function during nonneural organogenesis. PMID- 12636919 TI - Single-cell transcript analysis of pancreas development. AB - DNA microarray analysis was combined with a modified single-cell PCR procedure to study gene expression profiles of single cells at different stages of pancreatic development. This method identifies distinct cell types at embryonic day 10.5, a stage when the pancreatic epithelium is morphologically uniform. Some cells express unexpected combinations of genes, and these expression patterns provide new insights into pancreas development. Following on these findings, we use PCR products from different cell types to identify novel pancreatic genes, some of which mark subtypes of developing pancreatic cells. By integrating these data with previous genetic and biochemical studies, we propose a pathway for pancreatic cell development. This form of single-cell transcriptional analysis can be applied to any developmental process or tissue to characterize distinct cell types. PMID- 12636920 TI - Wnt3a plays a major role in the segmentation clock controlling somitogenesis. AB - The vertebral column derives from somites generated by segmentation of presomitic mesoderm (PSM). Somitogenesis involves a molecular oscillator, the segmentation clock, controlling periodic Notch signaling in the PSM. Here, we establish a novel link between Wnt/beta-catenin signaling and the segmentation clock. Axin2, a negative regulator of the Wnt pathway, is directly controlled by Wnt/beta catenin and shows oscillating expression in the PSM, even when Notch signaling is impaired, alternating with Lfng expression. Moreover, Wnt3a is required for oscillating Notch signaling activity in the PSM. We propose that the segmentation clock is established by Wnt/beta-catenin signaling via a negative-feedback mechanism and that Wnt3a controls the segmentation process in vertebrates. PMID- 12636922 TI - The cytoplasmic domain of Xenopus NF-protocadherin interacts with TAF1/set. AB - Protocadherins are members of the cadherin superfamily of cell adhesion molecules proposed to play important roles in early development, but whose mechanisms of action are largely unknown. We examined the function of NF-protocadherin (NFPC), a novel cell adhesion molecule essential for the histogenesis of the embryonic ectoderm in Xenopus, and demonstrate that the cellular protein TAF1, previously identified as a histone-associated protein, binds the NFPC cytoplasmic domain. NFPC and TAF1 coprecipitate from embryo extracts when ectopically expressed, and TAF1 can rescue the ectodermal disruptions caused by a dominant-negative NFPC construct lacking the extracellular domain. Furthermore, disruptions in either NFPC or TAF1 expression, using NFPC- or TAF1-specific antisense morpholinos, result in essentially identical ectodermal defects. These results indicate a role for TAF1 in the differentiation of the embryonic ectoderm, as a cytosolic cofactor of NFPC. PMID- 12636921 TI - Wg/Wnt signal can be transmitted through arrow/LRP5,6 and Axin independently of Zw3/Gsk3beta activity. AB - Activation of the Wnt signaling cascade provides key signals during development and in disease. Here we provide evidence, by designing a Wnt receptor with ligand independent signaling activity, that physical proximity of Arrow (LRP) to the Wnt receptor Frizzled-2 triggers the intracellular signaling cascade. We have uncovered a branch of the Wnt pathway in which Armadillo activity is regulated concomitantly with the levels of Axin protein. The intracellular pathway bypasses Gsk3beta/Zw3, the kinase normally required for controlling beta-catenin/Armadillo levels, suggesting that modulated degradation of Armadillo is not required for Wnt signaling. We propose that Arrow (LRP) recruits Axin to the membrane, and that this interaction leads to Axin degradation. As a consequence, Armadillo is no longer bound by Axin, resulting in nuclear signaling by Armadillo. PMID- 12636923 TI - SAS-4 is essential for centrosome duplication in C elegans and is recruited to daughter centrioles once per cell cycle. AB - The mechanisms governing centrosome duplication remain poorly understood. We identified a gene called sas-4 that is essential for this process in C. elegans. SAS-4 encodes a predicted coiled-coil protein that localizes to a tiny dot in the center of centrosomes throughout the cell cycle. FRAP experiments with GFP-SAS-4 transgenic embryos reveal that SAS-4 is recruited to the centrosome once per cell cycle, at the time of organelle duplication. Additional evidence indicates that SAS-4 is recruited to the daughter centriole or a closely associated structure. These findings identify SAS-4 recruitment as a key step in the centrosome duplication cycle. PMID- 12636924 TI - Capillary electrophoresis and its application in the clinical laboratory. AB - Over the past 10 years, capillary electrophoresis (CE) is an analytical tool that has shown great promise in replacing many conventional clinical laboratory methods, especially electrophoresis and high performance liquid chromatography (HPLC). The main attraction of CE was that it was fast, used small amounts of sample and reagents, and was extremely versatile, being able to separate large and small analytes, both neutral and charged. Because of this versatility, numerous methods for clinically relevant analytes have been developed. However, with the exception of the molecular diagnostic and forensic laboratories CE has not had a major impact. A possible reason is that CE is still perceived as requiring above-average technical expertise, precluding its use in a laboratory workforce that is less technically adept. With the introduction of multicapillary instruments that are more automated, less technique-dependent, in addition to the availability of commercial and cost effective test kit methods, CE may yet be accepted as a instrument routinely used in the clinical laboratories. Thus, this review will focus on the areas where CE shows the most potential to have the greatest impact on the clinical laboratory. These include analysis of proteins found in serum, urine, CSF and body fluids, immunosubstraction electrophoresis, hemoglobin variants, lipoproteins, carbohydrate-deficient transferrin (CDT), forensic and therapeutic drug screening, and molecular diagnostics. PMID- 12636925 TI - Polymorphisms in coagulation factor genes and their impact on arterial and venous thrombosis. AB - Arterial and venous thromboses, with their clinical manifestations such as stroke, myocardial infarction (MI), or pulmonary embolism, are the major causes of death in developed countries. Several studies in twins and siblings have shown that genetic factors contribute significantly to the development of these diseases. Since the advent of molecular genetics in medicine, it has been a focus of interest to elucidate the role of mutations in various candidate genes and their impact on hemostatic disorders such as arterial and venous thromboses. In this article, we review the current knowledge of the contribution of polymorphisms in coagulation factors to the development of thrombotic diseases. We show that in arterial thrombosis, results are controversial. Only for factor XIII 34Leu a protective effect on the development of myocardial infarction has been demonstrated in several studies. No other single polymorphism in a coagulation factor could be confirmed as a relevant risk factor, although there is evidence for a role of factor V Arg506Gln, factor VII Arg353Gln, and vWF Thr789Ala polymorphisms in patient subgroups. Further studies will be necessary to confirm the value of testing for genetic polymorphisms in arterial thrombosis. A large body of data is available on the role of factor V Arg506Gln and the prothrombin G20210A mutation in venous thrombosis. Some papers already recommend diagnosis and treatment strategies. We will discuss these recent publications on venous thrombosis in our review. PMID- 12636927 TI - Nitric oxide and S-nitrosothiols in human blood. AB - The hypothesis that endothelial-derived relaxing factor (EDRF) is nitric oxide has stimulated a wealth of research into the significance of this novel intriguing molecule. Given its short life, many storage forms of NO as well as targets have been postulated. Among these, a pool of derivatives of NO (S nitrosothiols, RSNOs) covalently bound to SH groups of proteins and low molecular weight thiols (e.g., glutathione) have been identified in various biological systems. The importance of RSNOs results from the very similar biological actions exhibited by both NO and RSNOs in vivo as well as in vitro. In particular, it has been observed that in the bloodstream, these molecules are able to provoke vasodilatation with a consequent fall in blood pressure and an antithrombotic effect by inhibition of platelet aggregation. Many hypotheses have been postulated about the biochemical species and the mechanisms involved in these processes, but many aspects have not yet been clarified. In addition, some RSNOs have been recently proposed to be clinical parameters, whose levels may vary under some pathological conditions. The therapeutic utility of RSNOs as an alternative to classic NO donors has also been suggested.Here, we provide a critical analysis of the main reports about the biochemical, physiological, pathological and therapeutic properties of RSNOs in the cardiovascular system. Particular attention is addressed to conflicting results and to discrepancies in the methodologies and models utilized. The numerous unanswered questions concerning the role of RSNOs in the control of vascular tone are discussed. PMID- 12636926 TI - The role of nutrition in preventing prostate cancer: a review of the proposed mechanism of action of various dietary substances. AB - BACKGROUND: Dietary modifications to prevent prostate cancer (PCa) continue to gain attention as research demonstrates that various dietary nutrients/supplements are related to decreased risk of developing prostate cancer (PCa). Several studies have focused on the antioxidant and nonantioxidant effects of various dietary substances in the prevention of PCa. Research into the mechanisms by which PCa is prevented, or its disease severity is reduced by dietary micronutrients and vitamins continues to enrich our understanding of the mechanisms by which PCa is initiated and progresses. METHODS: We reviewed the literature on dietary nutrients with antioxidant properties that have been shown to have a positive effect in reducing the incidence or preventing the occurrence of PCa including carotenoids (e.g., lycopene), retinoids (e.g., vitamin A), vitamin E, vitamin C, selenium, and polyphenols. Other nutrients examined included vitamin D and calcium. RESULTS: Many dietary micronutrients have demonstrated significant and complex effects on PCa cell proliferation, differentiation, and signaling related to the initiation, progression, and regression of PCa. CONCLUSION: Understanding the mechanisms by which various dietary nutrients exert their effects on PCa may make it possible to design effective drugs for treating PCa and to promote better nutrition and lifestyle changes in those at risk for PCa. PMID- 12636928 TI - Adrenal minerlocorticoids pathway and its clinical applications. PMID- 12636929 TI - Variability of post-methionine load plasma homocysteine assays. AB - BACKGROUND: Numerous variations of the methionine load test are frequently used as dynamic function tests to assess homocysteine metabolism. Lack of standardization impedes inter-laboratory comparisons. Criteria based on biological variation are suggested to standardize the methionine load test. METHODS: Weekly methionine load tests (n=5) with blood sampling at 0, 4, 6 and 8 h were performed on 15 young men. For both basal and post-methionine load homocysteine measurements, total variance (sigma(S)(2)), within-subject variance (sigma(I)(2)), between-subject variance (sigma(G)(2)) and analytical variance (sigma(A)(2)) were calculated from an appropriate analysis of variance (ANOVA). RESULTS: Plasma homocysteine concentrations measured 6 h after methionine loading had analytical, within-subject and between-subject coefficients of variation of 5.2%, 17.5% and 9.7%, respectively. Measurements at 4 h had a higher within subject coefficient of variation. Adjustment of post-methionine load homocysteine concentrations for basal levels resulted in considerable increases of all the measures of variation. CONCLUSIONS: Adjustment of post-methionine load plasma homocysteine concentrations for basal levels does not improve the interpretation of changes in serial results due to the higher analytical and biological variance of adjusted concentrations. It is suggested that the methionine load test is standardized to a single, unadjusted homocysteine measurement at 6 h. PMID- 12636930 TI - Lipid peroxidation and vitamin E in human coronary atherosclerotic lesions. AB - BACKGROUND: We aimed to assess the oxidant/antioxidant status within the ex vivo human coronary endarterectomy samples. METHODS: To achieve this, we measured products of lipid oxidation (malondialdehyde, 7-ketocholesterol), lipids (cholesterol, cholesteryl esters) and vitamin E in endarterectomy samples. RESULTS: Content of malondialdehyde in the plaque ranged from 0.23 to 37.36 microg/g. Unesterified cholesterol content ranged from 0.30 to 1.94 mg/g. It was 9.04+/-4.32% of total cholesterol. Total cholesterol content ranged from 1.73 to 23.69 mg/g. Cholesteryl palmitate content ranged from 0.57 to 19.10 mg/g, which is 11.43-60.86% of the total esters (mean+/-SD 40.27+/-18.42%). Cholesteryl oleate content ranged from 0.24 to 5.76 mg/g, being 9.97-21.81% of total esters (mean+/-SD 14.35+/-4.51%). Cholesteryl linoleate content ranged from 1.05 to 8.21 mg/g, being 17.84-45.15% of total esters (mean+/-SD 30.78+/-11.69%). Cholesteryl arachidonate content ranged from 0.51 to 4.20 mg/g, which is 7.56-22.87% of total esters (mean+/-SD 14.60+/-5.60%). The cholesteryl linoleate/cholesteryl oleate ratio (CL/CO) ranged from 1.01 to 4.33. Content of 7-ketocholesterol in the plaque ranged from 0.0 to 577.5 ng/g of wet weight. The 7-ketocholesterol/total cholesterol ratio was 0.003+/-0.003% (range from 0.0% to 0.008%). The 7 ketocholesterol/unesterified cholesterol ratio was 0.024+/-0.023% (range from 0.0% to 0.066%). The plaque content of vitamin E ranged from 0.0 to 40.9 microg/g of wet weight. CONCLUSION: The present study, comprising measurements of lipids, products of lipid peroxidation and vitamin E in 12 human coronary endarterectomy samples, lends the evidence for ongoing lipid peroxidation within an atherosclerotic lesion. PMID- 12636931 TI - Amniotic fluid levels of S100B protein in normal and trisomy-21 foetuses. AB - BACKGROUND: The human chromosome 21 has been shown to contain the gene for the beta subunit of the S100B protein. The present case-control study was aimed at investigating whether overproduction of S100B protein is detectable in the amniotic fluid of foetuses with trisomy-21. METHODS: Measurements of S100B in amniotic fluid samples from 14 pregnant women with trisomy-21 foetuses were compared with those obtained from 182 physiological pregnancies. S100B was measured in the samples using an immunoluminometric assay (LIA-mat Sangtec 100). RESULTS: Our results showed that S100B protein amniotic fluid levels were significantly higher in trisomy-21 foetuses (0.83+/-0.21 microg/l) than in controls (0.51+/-0.22 microg/l) (p<0.05). CONCLUSION: The present finding supports the notion that the expression of S100B is increased in trisomy-21 foetuses; it also constitutes a prerequisite basis for a possible involvement of the protein in pathogenic processes associated with trisomy-21, and/or for its potential employment as a diagnostic tool. PMID- 12636932 TI - Effect of vitamin D3 on the conversion of ethylene glycol to glycolate and oxalate in ethylene glycol-fed rats. AB - Hypercalciuria and hyperoxaluria are important risk factors in the pathogenesis of kidney stones. Urinary glycolate has also been reported to be elevated in patients with renal stones. 1,25-Dihydroxyvitamin D(3), the active metabolite of vitamin D, has been reported to induce hyperoxaluria after either oral or intravenous administration. 1-alpha-D(3), a synthetic derivative of vitamin D, together with ethylene glycol, has been reported to induce renal stones in experimental rats. We have examined the effect of 1-alpha-vitamin D(3) on urinary oxalate and glycolate excretion. Our results indicate that 1-alpha-D(3), together with ethylene glycol, caused a significant increase in urinary glycolate, without a parallel rise in urinary oxalate excretion, in ethylene glycol-fed rats. This increase in urinary glycolate was due to the synergistic effect of both drugs. PMID- 12636933 TI - High-molecular-mass isoform of aminopeptidase N/CD13 in serum from cholestatic patients. AB - BACKGROUND: Because non-denaturing electrophoresis and aminopeptidase activity staining often detect noncovalent multi-enzyme complexes, we adopted procedures to specifically detect the aminopeptidase N (APN) molecule itself in liver disease serum. METHODS: Sera or their immunoprecipitate with anti-APN monoclonal antibody were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) or two-dimensional electrophoresis and subsequent Western blotting with rabbit anti-APN serum. RESULTS: In all the patient sera examined, the 140-kDa APN isoform was predominant. In all the sera from 10 patients with cholestatic diseases (8 with extra-hepatic cholestasis and 2 with primary biliary cirrhosis), we observed the 260-kDa isoform that was immunoprecipitated with monoclonal APN antibodies and had a similar isoelectric point to the 140-kDa isoform. However, the 260-kDa isoform was observed faintly in 2 out of 12 patients with other liver diseases, including chronic hepatitis and cirrhosis. CONCLUSIONS: We found a novel high-molecular-mass APN isoform (260 kDa) in serum, which is highly likely to be a homodimer of APNs bound covalently and a promising marker of cholestasis. This suggests increased cross-linking reaction between two APN molecules in cholestatic patients. PMID- 12636935 TI - The lipoprotein lipase Ser447Ter mutation and risk of stroke in the Chinese. AB - BACKGROUND: Lipoprotein lipase (LPL) plays an important role in plasma lipoprotein metabolism. The Ser447Ter mutation of LPL may be associated with ischemic cerebrovascular diseases. We investigated whether the LPL variants were related to risk of strokes in Chinese Hans. METHODS: We recruited 160 patients with cerebrovascular diseases (ischemic stroke, n=96; hemorrhagic stroke, n=64) and 117 age-matched controls. All subjects were Chinese Hans. Subjects were analyzed for the Ser447Ter mutation by restriction fragment length polymorphisms of the LPL gene. RESULTS: As compared with controls, the frequency of LPL genotype CG (heterozygous Ser447Ter mutation) was lower in ischemic stroke patients (10.4% vs. 21.4%, p<0.05), and was not significantly different in hemorrhagic stroke patients (15.6% vs. 21.4%, p>0.05). The LPL G allele frequency was also lower in ischemic stroke patients (5.2%) vs. controls (10.7%, p<0.05). There was no difference between hemorrhagic stroke patients (7.8%) and controls. Serum Lp(a) concentrations were markedly lower in CG carriers than that in CC carriers in both stroke patients and the controls (p<0.05). There was no significant difference in the concentrations of other lipids. CONCLUSIONS: Patients with ischemic stroke have a lower frequency of the LPL Ser447Ter mutation, which indicates that this mutation may have protective effect on ischemic stroke. PMID- 12636934 TI - Modulation of extracellular homocysteine concentration in human cell lines. AB - BACKGROUND: Despite the growing evidence that plasma homocysteine is a cardiovascular risk factor, the mechanism behind the vascular injuries is still unknown. Information about the metabolism of homocysteine is, therefore, essential for an understanding of its role in atherogenesis, thereby enabling a modulation of that risk. METHODS: In the present study, we have examined the modulation of extracellular homocysteine in HeLa and hepatoma cell cultures in relation to a changed extracellular thiol redox status and in the presence of specific inhibitors of amino acid transporters. RESULTS: The findings in the present study show that a changed thiol redox status by copper ions, copper chelator or the monothiol, N-acetylcysteine (NAC), affects extracellular homocysteine in the same way in hepatoma cell cultures, but not to the same extent as observed in HeLa cell cultures. However, the dithiols, dithiothreitol (DTT) and alpha-lipoic acid (LA), which lowered extracellular homocysteine concentration in HeLa cell cultures, increased the extracellular total homocysteine concentration in hepatoma cell cultures, probably mainly as a result of increased release of homocysteine extracellularly. Studies with specific inhibitors of amino acid transporters in HeLa cell cultures showed that homocysteine uptake occurred mainly by system A and glutamate transporters. Hepatoma cells seemed to have a much smaller uptake capacity of homocysteine compared to HeLa cells. CONCLUSION: The lack of uptake capacity of homocysteine in hepatoma cells indicates that hepatocytes only play a small role in the elimination of homocysteine from circulation. Intracellular metabolism, cellular export and the complex pattern of homocysteine uptake in different cells are important to examine further in order to possibly be able to lower plasma homocysteine levels. PMID- 12636936 TI - Anti-inflammatory effect of Lonicera japonica in proteinase-activated receptor 2 mediated paw edema. AB - BACKGROUND: Lonicera japonica (Caprifoliaceae) has long been used for treatment of infectious diseases. In the present study, the anti-inflammatory effects of L. japonica water extract (AELJ) were investigated in proteinase-activated receptor 2 (PAR2)-mediated mouse paw edema. METHODS: Paw edema was induced by injection of trypsin or trans-cinnamoyl-LIGRLO-NH(2) (tc-NH(2)) into hindpaw of mice. AELJ (10, 50, 100, and 200 mg/kg) was orally administered 1 h before induction of inflammation. RESULTS: At doses of 50, 100 and 200 mg/kg, the AELJ showed significant inhibition of both change in paw thickness and vascular permeability. The AELJ (100 mg/kg) also significantly inhibited PAR2 agonists-induced myeloperoxidase (MPO) activity and tumor necrosis factor (TNF)-alpha expression in paw tissue. CONCLUSION: The present study demonstrated that AELJ has an anti inflammatory action for PAR2-mediated paw edema. PMID- 12636938 TI - Study on mechanism of action of Chinese medicine Chan Su: dose-dependent biphasic production of nitric oxide in trophoblastic BeWo cells. AB - BACKGROUND: Chan Su, a traditional Chinese medicine, is used for treating the heart diseases and other systemic illnesses. Our studies with animal model have revealed its role in increasing intracellular calcium concentration in cardiomyocytes. Nitric oxide (NO), a second messenger molecule, and its metabolites have been demonstrated to maintain and modulate multiple physiologic functions including the cardiovascular and reproductive systems. In order to explore the mechanism of action of Chan Su, we studied the ability of Chan Su to stimulate NO production in cultured trophoblastic BeWo cells. MATERIALS AND METHODS: BeWo cell is a cloned established cell line purified from human choriocarcinoma. These cells have some similarities in biological behavior with endothelial cells. Therefore, BeWo cell line may act as a model system for production of nitric oxide by Chan Su both in placenta and in cardiovascular tissue, and the results can easily be extrapolated to cardiomyocytes. Very small amount of ethanol extract of Chan Su was added to the cultured cells in KBM buffer and a chemiluminescence system was used for the measurement of nitric oxide. The amounts of Chan Su extract added to cultured cells were comparable to expected level of Chan Su in human serum after ingestion. We also repeated these experiments with bufalin, the active component of Chan Su. RESULTS: The ethanol extract of Chan Su (5 and 10 microg/ml) significantly increased NO production up to 110% of basal control value, but higher concentrations (40 and 80 microg/ml) of Chan Su (as expected in an overdose) resulted in decreased NO production below the control level. This biphasic effect on nitric oxide production was also observed with bufalin, the active component of Chan Su responsible for its digoxin-like immunoreactivity. The presence of bufalin in Chan Su preparation was confirmed by thin layer chromatography (TLC) analysis. CONCLUSIONS: Chan Su as well as bufalin is able to modulate the production of NO in BeWo cell line. PMID- 12636937 TI - Apolipoprotein epsilon 2/3 genotype and type III hyperlipoproteinemia among Taiwanese. AB - BACKGROUND: Type III hyperlipoproteinemia (HLP) is a genetic disorder of lipid metabolism in humans that predisposes affected subjects to the premature development of atherosclerosis. Most type III HLP occurs in homozygous carriers of apolipoprotein (apo) E2. The aims of this study were to determine the frequencies of different apo E genotypes in type III HLP in Taiwanese and to assess the possibility of apo E mutants in these patients. MATERIALS AND METHODS: Four hundred and seven patients with hyperlipoproteinemia were recruited. Electrophoresis, apo E genotyping and sequencing were performed. RESULTS: Of the 407 hyperlipoproteinemia, 8 were identified as type III HLP. In contrast to reports of high apo epsilon 2/2 genotype prevalence, only two of the type III HLP subjects were of the apo epsilon 2/2 genotype (25%). Fifty percent of the type III HLP were of apo epsilon 2/3 genotype. This observation was further reflected in a lower frequency of the epsilon 2 allele (0.563) and higher epsilon 3 allele (0.375) frequency. No rare apo E variant was found by direct sequencing. CONCLUSION: The most common genotype of type III HLP in Taiwan was apo epsilon 2/3 instead of apo epsilon 2/2. PMID- 12636940 TI - University medical schools: endangered species? PMID- 12636941 TI - Words and their integral role in scientific research. PMID- 12636942 TI - Comment on: "Peripheral utilization of glucose in pregnancy. III. Insulin tolerance". PMID- 12636943 TI - A randomized trial of the sleeved cytobrush and the endocervical curette. AB - OBJECTIVE: To compare endocervical specimens obtained with the endocervical curette to those obtained with the sleeved cytobrush. METHODS: All nonpregnant women undergoing either cervical conization or hysterectomy were eligible for this randomized, split-sample trial. Both endocervical curette and cytobrush sampling were performed on all 62 participants before surgery, with randomization designating the order of the sampling procedures. A pathologist blinded to sampling order reviewed study specimens. The endocervical canals of the surgical specimens were evaluated against endocervical curette and brush samples. Odds ratios (ORs), relative risks (RRs), and risk differences were used to compare the sensitivity and specificity of each procedure in unmatched and matched analyses. RESULTS: There was a significantly higher rate of inadequate specimens in the endocervical curette group (22% versus 2% in the brush group; 95% confidence interval [CI] for the difference: 9%, 31%). Unmatched analysis showed the two tests to be comparable in terms of sensitivity and specificity. Whereas the specificities of both tests were high (100% for the endocervical curette, 88% for the brush; RR 1.13, 95% CI 1.00, 1.28), the sensitivities were poor (44% for the sleeved brush and 32% for the endocervical curette; RR 1.38, 95% CI 0.65, 2.94). Matched analysis showed the sleeved endocervical brush to be a more sensitive sampling method compared with the curette (OR 2.04, 95% CI 0.98, 4.22). Sparse data prevented a matched analysis on the specificity of the two tests. CONCLUSION: Endocervical sampling with the sleeved cytobrush achieves similar sensitivity and specificity to that of traditional endocervical curettage. Given the much greater rate of inadequate specimens when using the endocervical curette, the sleeved cytobrush is a reasonable alternative to this technique. PMID- 12636944 TI - Self-reported heavy bleeding associated with uterine leiomyomata. AB - OBJECTIVE: To characterize the relationship between self-reported bleeding symptoms and uterine leiomyoma size and location. METHODS: The leiomyoma status of a randomly selected sample of women aged 35-49 in the Washington, DC, area was determined using abdominal and transvaginal ultrasound to measure size and location of leiomyomata found at screening. Women were asked about symptoms of heavy bleeding (gushing-type bleeding, long menses, pad/tampon use) in a telephone interview. Using multivariable regression, we examined the relationships between leiomyoma characteristics and heavy bleeding symptoms among 910 premenopausal women. RESULTS: Women with leiomyomata (n = 596) were more likely to report gushing-type bleeding than women without leiomyomata; risk increased with leiomyoma size. Adjusted relative risks with 95% confidence intervals (CI) for women in each leiomyoma size category compared with the reference category (women without leiomyomata) were as follows: adjusted relative risk of 1.4 (95% CI 1.1, 1.9) for diffuse only, adjusted relative risk of 1.4 (95% CI 1.1, 1.8) for small leiomyomata (less than 2 cm), adjusted relative risk of 1.6 (95% CI 1.3, 2.0) for medium leiomyomata (2-5 cm), and adjusted relative risk of 1.9 (95% CI 1.5, 2.5) for large leiomyomata (greater than 5 cm). Reported use of eight or more pads/tampons on the heaviest days of menstrual bleeding increased with leiomyoma size, with a nearly 2.5-fold risk for women with large leiomyomata compared with women without leiomyomata (adjusted relative risk of 2.4; 95% CI 1.8, 3.1). Nonsubmucosal leiomyomata were associated with essentially the same increase in heavy bleeding as submuscosal leiomyomata of similar size. CONCLUSION: Small leiomyomata were associated with increased risk of heavy bleeding, and risk increased with size. Contrary to published articles, nonsubmucosal leiomyomata were associated with heavy bleeding to the same extent as submucosal leiomyomata. PMID- 12636945 TI - Immunogenetic correlates for Chlamydia trachomatis-associated tubal infertility. AB - OBJECTIVE: To understand immunogenetic mechanisms of Chlamydia trachomatis infection and tubal scarring. METHODS: We measured and compared previously significant human leukocyte antigen (HLA) class II DQ alleles, their linked DRB genes, and polymorphisms in selected cytokine genes (tumor necrosis factor alpha 308 promoter; transforming growth factor beta1-10 and -25 codons; interleukin 10 1082, -819, and -592 promoters; interleukin 6-174 promoter; and interferon gamma+874 codon 1) among Kenyan women with confirmed tubal infertility with and without C trachomatis microimmunofluorescence antibody. RESULTS: Two class II alleles, HLA-DR1*1503 and DRB5*0101, were detected less commonly in C trachomatis microimmunofluorescence seropositive women than in C trachomatis microimmunofluorescence seronegative women with infertility (0% versus 20%; odds ratio [OR] 0.05; 95% confidence interval [CI] 0, 0.7, and 6% versus 26%; OR 0.2; 95% CI 0.02, 1.0, respectively). These alleles are commonly linked as a haplotype at the DRB locus. This finding could not be explained through linkage disequilibrium with the other studied HLA or cytokine genes. CONCLUSION: These alleles may lead to an immunologically mediated mechanism of protection against C trachomatis infection and associated tubal damage, or alternatively increase risk for tubal scarring due to another cause. PMID- 12636946 TI - Endometrial cancer: the potential role of cervical cytology in current surgical staging. AB - OBJECTIVE: To evaluate cervical cytology, tumor grade from endometrial sampling, and myometrial invasion with the risk of nodal spread in endometrial cancer. METHODS: Cervical cytology was obtained in 300 patients with endometrial cancer before surgical staging, which included lymphadenectomy. Tumor grade and histology from endometrial sampling were compared with final pathology, and the risk of nodal spread in relation to cervical cytology, tumor grade, and myometrial invasion was assessed using chi(2) and logistic regression analysis. RESULTS: Endometrial cells on cervical cytology, deep myometrial invasion, and high-grade tumor were associated with 91%, 87%, and 83% of the cases with nodal spread, respectively. In patients with grade 1 tumor on biopsy, final pathology revealed grade 2 in 21%, and grade 3 in 2%. In patients with normal cervical cytology, no nodal metastases occurred with grade 1 tumor on biopsy, and no aortic metastases occurred, regardless of grade. Cervical cytology and tumor grade contributed independently to the likelihood of nodal metastases. CONCLUSION: All patients with endometrial cancer should undergo lymphadenectomy until a reliable system is found to identify those with negligible (less than 1%) risk of nodal spread. The risk of lymph node spread in those with normal cervical cytology is low (2%). Further study of those with normal cervical cytology is needed to determine if lymphadenectomy can be omitted with grade 1 tumor on biopsy, or whether aortic lymphadenectomy is necessary regardless of grade. PMID- 12636947 TI - Incorporation of inhibin-A in second-trimester screening for Down syndrome. AB - OBJECTIVE: To evaluate the efficacy of the second-trimester quadruple test (maternal serum alpha-fetoprotein, unconjugated estriol, human chorionic gonadotropin, and inhibin-A) in prenatal screening for Down syndrome. METHODS: All quadruple tests performed on singleton pregnancies over a 32-month period were reviewed. The sensitivity and false-positive rates were compared with the theoretic rates predicted by the screening model when applied to a population of women with the same maternal age distribution. RESULTS: Twenty-three thousand seven hundred four women with unaffected pregnancies and 45 women with Down syndrome-affected pregnancies received the quadruple test. Mean analyte values for both unaffected and affected pregnancies were similar to those expected. The sensitivity of the quadruple test, which was based on ascertainment of all viable affected pregnancies in the screened population, was 85.8%. This sensitivity did not significantly differ from an expected 83.8% (P =.8). The initial false positive rate, 9.0%, was significantly below that expected (9.9%) (P =.002) and was further reduced to 8.2% after correction for major gestational age errors. The positive predictive value of the quadruple test was one in 51. Women with an affected pregnancy and a positive test result (true positives) generally had very high risks (median 1:22) relative to women with false-positive results (median risk 1:111). CONCLUSION: The quadruple test meets or exceeds performance expectations and appears to represent an improvement over the widely used triple test. PMID- 12636948 TI - Effects of unfractionated and low molecular weight heparin on antiphospholipid antibody binding in vitro. AB - OBJECTIVE: To compare the efficacy of unfractionated heparin and low molecular weight heparin in the in vitro binding of antiphospholipid antibodies obtained from the sera of patients with recurrent pregnancy loss. METHODS: Women with immunoglobulin (Ig) G antibodies to the phospholipids cardiolipin and phosphatidylserine were selected based on a positive test by a standard enzyme linked immunosorbent assay (ELISA). The sera were reassayed for antiphospholipid antibodies in a modified ELISA using increasing doses of unfractionated heparin or low molecular weight heparin (0, 16, 32, 64, 128, and 256 IU). Sera were fractionated by unfractionated and low molecular weight heparin affinity chromatography to compare the binding avidity and antiphospholipid antibody activity. RESULTS: All sera demonstrated a dose-dependent inhibition in measured antiphospholipid antibody activity with the addition of unfractionated or low molecular weight heparin. Levels of IgG cardiolipin and IgG phosphatidylserine were significantly inhibited in the presence of 32 IU of low molecular weight heparin (P <.001 and P <.05, respectively) and in the presence of 64 IU of unfractionated heparin (P <.001 and P <.05, respectively). Antiphospholipid antibody binding activity in serum as measured in the ELISA was maximally reduced 76-89% with 256 IU of either heparin derivative. Affinity chromatography with unfractionated or low molecular weight heparin columns absorbed 72% and 66% of IgG cardiolipin activity, respectively, and 46% and 54% of IgG phosphatidylserine activity, respectively. CONCLUSION: These data suggest that low molecular weight heparin and unfractionated heparin reduce the in vitro binding of antiphospholipid antibodies on a per unit basis. Both heparins demonstrate binding activity similar to that of antiphospholipid antibodies in vitro. PMID- 12636949 TI - Early second-trimester individualized estimation of trisomy 18 risk by ultrasound. AB - OBJECTIVE: To report two second-trimester ultrasound algorithms for trisomy 18 prediction. METHODS: Femur length, gross anomaly, choroid plexus cysts, two vessel cord, and maternal age were documented in pregnancies undergoing genetic amniocentesis. Stepwise logistic regression was used to identify 1) the significant markers for predicting trisomy 18 when gross anomaly was not considered (algorithm 1) and 2) when gross anomaly was also considered (algorithm 2). Patient-specific risk was calculated based on the significant ultrasound markers plus maternal age. The diagnostic accuracy of each algorithm was determined. RESULTS: There were 1167 normal and 47 trisomy 18 cases. The mean gestational ages were 16.5 weeks (standard deviation [SD] 1.5) and 17.9 weeks (SD 1.6), respectively. Algorithm 1 consisted of maternal age, choroid plexus cyst, femur length, and two-vessel cord. The sensitivity and false positive rates were 61.7% and 1.5%, respectively, with an area under the receiver operating characteristics curve of 0.880 (P <.001). Algorithm 2 (age, femur length, gross anomaly, and choroid plexus cyst) had a sensitivity of 72.3% and false positive rate of 0.9% with an area under the curve of 0.956 (P <.001). Comparable detection rates were achieved in early gestation at up to and including 17.5 weeks (72.4% and 82.8%, algorithms 1 and 2, respectively, at a 4.0% false positive rate). CONCLUSION: The ultrasound markers were sensitive for trisomy 18 detection in the early second trimester. PMID- 12636950 TI - A randomized trial of docosahexaenoic acid supplementation during the third trimester of pregnancy. AB - OBJECTIVE: To hypothesize that higher intake of docosahexaenoic acid, an n-3 long chain polyunsaturated fatty acid, would increase duration of gestation and birth weight in US women. METHODS: This was a randomized, double-blind, controlled, clinical trial. Subjects were enrolled in an ambulatory clinic where they received prenatal care. This was a population-based sample. Most subjects received government assistance for medical care and most were black (73%). Subjects were enrolled between the 24th and 28th week of pregnancy and consumed docosahexaenoic acid (33 or 133 mg) from eggs until parturition. Gestational age and birth weight were the main study outcomes. Infant length and head circumference, preterm birth, and low birth weight were secondary outcomes. RESULTS: Eighty-three percent of subjects completed the study (291 of 350 enrolled). No subject was discontinued for an adverse event. After controlling for important predefined risk factors and confounding variables, gestation increased by 6.0 +/- 2.3 days (P =.009) in the higher docosahexaenoic acid group. Birth weight, length, and head circumference increased, but did not reach statistical significance (P =.06-.18), although the increases could be clinically important indications of enhanced intrauterine growth. No safety concerns were raised by the study. CONCLUSION: Duration of gestation increased significantly when docosahexaenoic acid intake was increased during the last trimester of pregnancy. The increase in gestation was similar to that reported for interventions with much larger amounts of n-3 long chain polyunsaturated fatty acids. PMID- 12636951 TI - Triage of atypical squamous cells of undetermined significance with hybrid capture II: colposcopy and histologic human papillomavirus correlation. AB - OBJECTIVE: To estimate the effectiveness of Hybrid Capture II to predict high grade cervical intraepithelial neoplasia (CIN) from a cytological cervical sample. Evidence of high-risk human papillomavirus (HPV) was also determined from biopsy samples using the polymerase chain reaction (PCR) for women referred with atypical squamous cells of undetermined significance (ASCUS) Papanicolaou smears. METHODS: We screened 8170 women with Papanicolaou smears, of whom 278 (3.4%) returned ASCUS. All ASCUS cases underwent colposcopy and Hybrid Capture II testing. High-grade CIN biopsy specimens were tested for high-risk HPV by PCR. RESULTS: Nearly 30% of ASCUS cases had CIN biopsy results (11.9% showing CIN II or CIN III and 17.6% showing CIN I). Hybrid Capture II positive rates were 93.3% for cases with CIN III, 72.2% for CIN II, and 51.0% for CIN I (P <.001). ASCUS Premalignant Process Favored cases showed a 28.1% high-grade biopsy rate and a 100% Hybrid Capture II positive rate. For ASCUS-Undefined and ASCUS-Reactive Process Favored cases, Hybrid Capture II returned positive in 90.9% of CIN III and 61.5% of CIN II cases (P <.001). Sixty-nine of 178 (38.8%) patients with no evidence of CIN tested positive for Hybrid Capture II. Human papillomavirus deoxyribonucleic acid (DNA) high-risk subtypes were detected by PCR in the tissue of all high-grade CIN cases with negative Hybrid Capture II results. CONCLUSION: Hybrid Capture II returned negative in 25% of cases with biopsy-proven high-grade CIN with associated high-risk HPV DNA by PCR (non-Premalignant ASCUS subset), and positive in 39.3% of cases with normal results; this limits its clinical utility. PMID- 12636952 TI - Predictors of Papanicolaou smear return in a hospital-based adolescent and young adult clinic. AB - OBJECTIVE: Sexually active young women have relatively high rates of abnormal cervical cytology, yet compliance with return for Papanicolaou smear screening and follow-up appointments is poor. The aim of this study was to determine whether a theory-based model could explain compliance with return visits. METHODS: Participants in this longitudinal cohort study were sexually active young women 12-24 years of age presenting to a hospital-based adolescent clinic. Participants completed self-administered surveys and were then followed for up to 15 months to assess for the outcome measure, return. Logistic regression modeling was used to determine variables independently associated with return. RESULTS: The outcome measure, return, was available for 439 of 490 participants (90%). Mean participant age (+/- standard deviation) was 18.3 (+/- 2.2) years, 49% were black, 23% were Hispanic, and 51% had Medicaid health insurance. Variables independently associated with return included belief that the Papanicolaou smear will not be painful (odds ratio [OR] 1.73, 95% confidence interval [CI] 1.08, 2.83), belief that return for follow-up will prevent cervical cancer (OR 1.83, 95% CI 1.12, 3.07), likelihood that the doctor will be honest (OR 4.07, 95% CI 1.37, 17.5), and low self-reported impulsivity (OR 1.66, 95% CI 1.06, 2.63). Family history of cervical cancer was associated with decreased likelihood of return (OR 0.28, 95% CI 0.08, 0.78). CONCLUSION: Specific beliefs about Papanicolaou smears and providers, low self-reported impulsivity, and no family history of cervical cancer are associated with return for Papanicolaou smear screening and follow-up visits. These findings may guide the design of interventions to increase compliance with recommendations for Papanicolaou smear return. PMID- 12636953 TI - The tensile strength of uterosacral ligament sutures: a comparison of vaginal and laparoscopic techniques. AB - OBJECTIVE: To compare the tensile strength of two approaches for uterosacral ligament suturing using a cadaver model. METHODS: In 12 unembalmed cadavers, four polytetrafluoroethylene sutures were placed through the uterosacral ligaments. In each cadaver, two sutures were placed laparoscopically, and two more were placed vaginally. A single, experienced surgeon placed all laparoscopic sutures (n = 23), and another experienced surgeon placed all vaginal sutures (n = 22). A blinded team of investigators measured the distance from each suture to the ipsilateral ischial spine; determined whether any sutures incorporated ureters, viscera, or large vessels; and then passed the sutures through an apical vaginal incision. Using a hand-held tensiometer, progressive tensile load was then applied to these sutures along the axis of the vagina until they either broke or were completely dislodged from the ligaments. RESULTS: The average peak tension required to break or dislodge the sutures was 26.2 +/- 8.8 psi (laparoscopic) and 22.5 +/- 7.4 psi (vaginal) (P =.14, 95% confidence interval [CI] -1.2, 8.6). The average force required for suture breakage (n = 28) was 28 +/- 7 psi, and the average force applied when ligament failure occurred (n = 17) was 18.5 +/- 6 psi (P <.001, 95% CI -13.8, -5.2). The average distance from a laparoscopic or vaginal suture to the ipsilateral ischial spine was 19.1 +/- 7 mm and 17.4 +/- 6 mm, respectively (P =.46, 95% CI -3.0, 6.4). None of the sutures from either technique were found to incorporate a visceral structure, ureter, or great vessel. CONCLUSION: These suturing techniques appear to be equal in tensile strength. PMID- 12636954 TI - The effects of peer counseling on smoking cessation and reduction. AB - OBJECTIVE: To evaluate a peer counseling intervention for pregnant smokers. METHODS: One hundred forty-two pregnant, predominantly Hispanic women were assigned to a peer-led smoking cessation program or to usual care. RESULTS: Compared with usual care, peer counseling reduced smoking (-9.1 versus -4.5 cigarettes daily, P =.03), but did not affect absolute quit rates (24% versus 21%) at 36 weeks' gestation. Infant birth weight negatively correlated with cigarettes smoked per day (r = -0.29, P <.01) and expired carbon monoxide (r = 0.39, (P <.001) at delivery. Birth weight for infants born to women who quit smoking averaged 7.2 lb versus 6.8 and 6.3 lb for mothers smoking one to six and more than six cigarettes per day at delivery (P <.01). CONCLUSION: Peer counseling reduced the number of cigarettes smoked daily but did not increase cigarette abstinence rates. Infant birth weight increases with both smoking cessation and smoking reduction, suggesting that peer counseling intervention programs may improve newborn health despite their failure to affect smoking cessation. PMID- 12636955 TI - Acute pulmonary edema in pregnancy. AB - OBJECTIVE: To describe the incidence, predisposing conditions, and inciting factors culminating in pulmonary edema in the pregnant patient. METHODS: A review of 62,917 consecutive pregnancies delivered at our institution from January 1, 1989 to June 1, 1999 was undertaken for the diagnosis of pulmonary edema. Each chart was reviewed for maternal demographics, admission diagnoses, medication use, gestational age at diagnosis, fluid balance, coexisting maternal illness, tocolytic use, evidence of preeclampsia, and diagnostic criteria. After careful review of the records, the most likely cause of pulmonary edema was assigned. RESULTS: Fifty-one women (0.08%) were diagnosed with acute pulmonary edema during pregnancy or in the postpartum period. The mean patient age at the time of diagnosis was 27.6 +/- 6.4 years. The mean gestational age at the time of diagnosis was 31.5 +/- 6.8 weeks. The diagnosis of pulmonary edema was made during the antepartum period in 24 patients (47%), the intrapartum period in seven (14%), and the postpartum period in 20 (39%). The most common attributable causes were tocolytic use (13 patients [25.5%]), cardiac disease (13 patients [25.5%]), fluid overload (11 patients [21.5%]), and preeclampsia (nine patients [18%]). Those with fluid overload identified as the likely etiology had a significantly greater mean positive fluid balance (6022 +/- 3340 mL). All patients whose pulmonary edema was secondary to tocolytic use received multiple simultaneous tocolytic agents; the most common combination was intravenous magnesium sulfate and subcutaneous terbutaline. Six of the 13 women with cardiac disease were found to have previously undiagnosed structural heart disease. CONCLUSION: The most common causes of pulmonary edema are the use of tocolytic agents, underlying cardiac disease, fluid overload, and preeclampsia. PMID- 12636956 TI - Intravaginal clindamycin to reduce preterm birth in women with abnormal genital tract flora. AB - OBJECTIVE: To assess the ability of clindamycin vaginal cream to reduce the incidence of preterm birth in women with abnormal genital tract flora in the second trimester of pregnancy. METHODS: This was a randomized, double-blind, placebo-controlled, tricenter study. A total of 409 women with abnormal genital tract flora on Gram stain of vaginal secretions at 13-20 weeks' gestation were randomized to receive a 3-day course of clindamycin vaginal cream or placebo. Those women who still had abnormal vaginal flora 3 weeks later received a 7-day course of the original study drug (ie, either clindamycin vaginal cream or placebo as per original randomization). The primary outcome measure was the incidence of preterm birth. RESULTS: There was a statistically significant reduction in the incidence of preterm birth in the clindamycin vaginal cream group (4%) compared with placebo (10%) (P <.03). Significantly more babies born preterm (63%) required admission to the neonatal intensive care unit compared with term infants (4%) (P <.001). CONCLUSION: A 2% clindamycin vaginal cream, when compared with placebo administered to women with abnormal genital tract flora before 20 weeks' gestation, can reduce the incidence of preterm birth by 60% and hence the need for neonatal intensive care. PMID- 12636957 TI - Correlation of elevated leptin levels in amniotic fluid and maternal serum in neural tube defects. AB - OBJECTIVE: To measure maternal serum and amniotic fluid leptin concentrations in pregnant women diagnosed antenatally as having fetuses with a neural tube defect in the second trimester. METHODS: Twenty pregnant women who had fetuses with a neural tube defect detected on ultrasonography (neural tube defect group) in the second trimester and 20 women who had abnormal triple screens indicating an increased risk for Down syndrome but had healthy fetuses (control group) were enrolled in the study. Amniotic fluid was obtained by amniocentesis, and maternal serum samples were taken simultaneously. RESULTS: The mean leptin levels in amniotic fluid (P <.001) and maternal serum (P <.05) of patients who had fetuses with a neural tube defect were found to be significantly higher than control group levels. The mean leptin levels in maternal serum of both groups were also higher than leptin levels in amniotic fluid (P <.05 for the neural tube defect group and P <.001 for the control group). Although there were significant correlations between maternal weight, weight gain, body mass index at the time of amniocentesis, and maternal serum leptin concentrations in both groups, a significant correlation between leptin concentrations in maternal serum and amniotic fluid was found only in the neural tube defect group (P <.05). CONCLUSIONS: We found significantly higher leptin levels in both amniotic fluid and maternal serum of patients who had fetuses with a neural tube defect. We suggest that the main source of leptin in amniotic fluid of pregnant women who had fetuses with a neural tube defect is the leakage into amniotic fluid from cerebrospinal fluid. The increase of maternal serum leptin concentrations has been attributed to the transportation of amniotic fluid leptin to the maternal circulation. PMID- 12636958 TI - Intrauterine exposure to preeclampsia and adolescent blood pressure, body size, and age at menarche in female offspring. AB - OBJECTIVE: To investigate whether female offspring of preeclamptic pregnancies have higher blood pressure, lower height, higher body mass index (BMI), and later age at menarche compared with offspring of normotensive pregnancies. METHODS: Questionnaire information on age at menarche and measurements of blood pressure, height, and weight were collected among 4096 Norwegian girls 13-19 years old. Individual linkage to perinatal data registered at the national Medical Birth Registry allowed us to study the relationship of preeclampsia in the mother with adolescent blood pressure, body size, and age at menarche of daughters. RESULTS: Maternal preeclampsia was associated in the female offspring with higher systolic (2.9 mm Hg difference, P <.001) and diastolic (1.7 mm Hg difference, P =.001) blood pressure during adolescence and higher weight (3.4 kg difference, P <.001) and BMI (22.6 versus 21.5, P <.001). After adjustment for adolescent BMI, the difference in systolic blood pressure was attenuated from 2.9 to 1.7 mm Hg (P =.017), and from 1.7 to 0.9 mm Hg (P =.08) for diastolic blood pressure. CONCLUSION: Intrauterine exposure to preeclampsia was associated with increased adolescent blood pressure. The association may be causally related to adult hypertension but could also be confounded by higher BMI during adolescence. PMID- 12636959 TI - Adolescents' contraceptive use and pregnancy history: is there a pattern? AB - OBJECTIVE: To examine the association between pregnancy experience and adolescents' contraceptive use. METHODS: We conducted a retrospective study of 920 sexually active adolescents not desiring pregnancy. Adjusted multivariable logistic regression analyses were used to assess the explanatory value of previous birth and abortion as well as first pregnancy at presentation on contraceptive practice. RESULTS: Twenty-seven percent of the adolescents had been pregnant. Regardless of pregnancy history, 52% of adolescents used noneffective contraception (ie, condoms inconsistently or no method). Adolescents with previous abortion were three times (odds ratio [OR] 3.4, 95% confidence interval [CI] 1.6, 7.3) more likely than never-pregnant adolescents to use hormonal contraception. However, although more likely to use contraceptive injections or implants, adolescent mothers were not more likely than never-pregnant adolescents to use oral contraceptives. Adolescents with prior abortion or birth were less than half (ORs 0.3 and 0.4, 95% CIs 0.2, 0.5 and 0.2, 0.6, respectively) as likely as never-pregnant adolescents to use condoms consistently. CONCLUSION: Previous pregnancy is associated with increased likelihood to use hormonal methods; however, a significant proportion of adolescents use noneffective methods and, thus, are at risk for repeat teenage pregnancies. PMID- 12636960 TI - Contraceptive effectiveness of a polyurethane condom and a latex condom: a randomized controlled trial. AB - OBJECTIVE: To compare the contraceptive effectivenesses of a polyurethane condom and a standard latex condom. Secondary outcomes of interest were safety, functionality, discontinuation, and acceptability. METHODS: We randomized 901 couples to use either the polyurethane condom or a standard latex condom as their only form of contraception. We tested for pregnancy at enrollment and at every scheduled follow-up visit (weeks 4, 10, 16, 22, and 30). RESULTS: The 6-month typical-use pregnancy probabilities were 9.0% (95% confidence interval [CI] 5.9, 12.2) for the polyurethane group and 5.4% (95% CI 2.9, 7.8) for the latex group; the hazard ratio was 1.7 (95% CI 1.1, 2.7), and we failed to reject the null hypothesis of our test of noninferiority. Females in the polyurethane group reported fewer genital irritations (hazard ratio 0.6; 95% CI 0.5, 0.8; P <.01), whereas males in both groups reported the same number of genital irritations (hazard ratio 1.0; 95% CI 0.7, 1.5; P =.94). Total clinical failures (breakage and slippage) were 8.4% for the polyurethane condom and 3.2% for the latex condom (difference 5.3%, 90% CI 2.8, 7.7). The risk of discontinuation did not differ between groups. Participants judged both condoms favorably in terms of the four primary acceptability outcomes (willingness to purchase, willingness to recommend, confidence in method, and general comfort). CONCLUSION: The polyurethane condom was not shown to be as effective as the latex comparator condom for pregnancy prevention. However, the risk of pregnancy in the polyurethane group falls in the range of other barrier methods. For people with latex sensitivity or who find latex condoms unacceptable, this polyurethane condom represents one of several synthetic male condom alternatives currently available on the US market. PMID- 12636962 TI - Maternal serum corticotropin-releasing hormone at midgestation in Hispanic and white women. AB - OBJECTIVE: Maternal circulating corticotropin-releasing hormone analysis at midgestation has been proposed as a parameter for the prediction of preterm birth. However, one recent study has reported that corticotropin-releasing hormone concentrations at midgestation differ in the black and white populations. These findings led us to investigate whether other populations have differing concentrations of maternal circulating corticotropin-releasing hormone that may require reference to specific population-based medians for optimal midgestational screening. METHODS: In this study we have defined the mean and median concentrations of maternal circulating corticotropin-releasing hormone in Hispanic and white populations at each gestational week from 14 to 18 weeks of pregnancy, using a sensitive and specific radioimmunoassay. RESULTS: Corticotropin-releasing hormone concentrations were found to be significantly lower in the Hispanic population as compared with whites at 16, 17, and 18 weeks' gestation. The distribution of corticotropin-releasing hormone, expressed as multiples of the median (MoM) using the appropriate ethnicity-related median, was estimated for each gestational week and for each population. No differences were observed in the distribution of the ethnicity-adjusted MoM for Hispanics and whites. CONCLUSION: These data demonstrate that ethnicity is a significant factor affecting corticotropin-releasing hormone concentrations at midgestation in the Hispanic and white populations. The use of ethnicity-specific medians to estimate the ethnicity-specific MoM for the corticotropin-releasing hormone concentrations may enhance the predictive value of midgestational maternal corticotropin releasing hormone as a screening parameter for the prediction of preterm birth. PMID- 12636961 TI - Incident and persistent vulvovaginal candidiasis among human immunodeficiency virus-infected women: Risk factors and severity. AB - OBJECTIVE: To examine risk factors for vulvovaginal candidiasis among women with or at risk for human immunodeficiency virus (HIV) infection. METHODS: Data were from 856 HIV-infected women and 421 at-risk uninfected women observed semiannually at four study sites from April 1993 through February 1999. At enrollment women were 15-55 years old and had no acquired immunodeficiency syndrome-defining conditions. Three definitions for vulvovaginal candidiasis of differing severity were constructed using data from vaginal Candida culture and Gram stains scored for yeast and three signs on pelvic examination (vulvovaginal edema, erythema, or discharge): 1) culture or Gram stain positivity plus at least one clinical sign, 2) culture or Gram stain positivity plus at least two clinical signs, and 3) visible yeast on Gram stain plus at least one clinical sign. RESULTS: The prevalence and cumulative incidence of each definition of vulvovaginal candidiasis were greater among HIV-infected women than among women not infected with HIV (P <.01 for all comparisons). Stratified by status at the preceding visit, vulvovaginal candidiasis was most likely among women with prior vulvovaginal candidiasis, least likely among women without earlier Candida colonization, and intermediately likely among women with preceding subclinical Candida colonization. Among HIV-infected women, lower CD4 count and higher HIV viral load were associated with vulvovaginal candidiasis. Several other factors were independently associated with vulvovaginal candidiasis, with strong associations for diabetes mellitus and pregnancy in particular. Vulvovaginal candidiasis was not more severe among HIV-infected women. CONCLUSION: Vulvovaginal candidiasis occurred with higher incidence and greater persistence, but not greater severity, among HIV-infected women. PMID- 12636963 TI - Factors associated with success of emergent second-trimester cerclage. AB - OBJECTIVE: To assess the factors associated with delivery greater than or equal to 28 weeks' gestation after placement of an emergent cerclage in women with singleton gestations. METHODS: All women who underwent emergent cerclage, defined as any cerclage placed between 16 and 24 6/7 weeks' gestation in response to documented cervical change on physical examination, at Northwestern Memorial Hospital from 1980 to 2000 were identified. Univariable and multivariable analyses were used to determine the factors most associated with achieving at least 28 weeks' gestation. RESULTS: One hundred sixteen women were eligible for analysis. Maternal age, race, and operative variables such as suture type and use of antibiotics were not associated with differences in the frequency of delivery at or after 28 weeks. Cerclage placement at or after 22 weeks' gestation increased the likelihood of reaching 28 weeks, whereas several cervical examination findings (dilatation greater than 3 cm, cervical length less than 0.5 cm, and membranes prolapsing beyond the external cervical os) as well as need for placement in a nullipara significantly reduced the likelihood of reaching 28 weeks. In multivariable analysis, nulliparity (odds ratio 0.31, 95% confidence interval 0.1, 0.8) and membranes prolapsing beyond the external cervical os (odds ratio 0.24, 95% confidence interval 0.1, 0.4) continued to be associated with delivery before 28 weeks, whereas cerclage placement at or after 22 weeks (odds ratio 3.2, 95% confidence interval 1.2, 8.6) increased the chance of achieving at least 28 weeks' gestation. CONCLUSION: Nulliparity, the presence of membranes prolapsing beyond the external cervical os, and gestational age less than 22 weeks at cerclage placement are associated with decreased chance of delivery at or after 28 weeks after emergent cerclage; these factors may be used to help counsel patients considering the procedure. PMID- 12636964 TI - The effect of birth weight discordance on twin neonatal mortality. AB - OBJECTIVE: To estimate the association between birth weight discordance and neonatal mortality controlling for the effects of fetal growth, and to understand the differences in the incidence of mortality between larger and smaller infants. METHODS: This analysis is based on the National Center for Health Statistics matched multiple birth data set file containing all twin births in the United States from 1995 through 1997. Birth weight discordance was grouped into four levels (15-19%, 20-24%, 25-29%, and 30% or more). Generalized estimating equations were used to obtain adjusted odds ratios and 95% confidence intervals to estimate the mortality risk associated with discordance after adjusting for fetal growth. RESULTS: Mortality was 11 times higher among highly discordant smaller twins (30% or more) compared with nondiscordant smaller twins (43.4 and 3.8 per 1000, respectively). Risk estimates ranged from 1.08 (95% confidence interval 0.85, 1.38) among 15-19% discordant twins to 2.05 (95% confidence interval 1.66, 2.51) among 30% or more discordant twins. Larger twins had similar risk estimates. After accounting for the association between fetal growth and discordance, mortality risk was substantially higher among smaller and larger twins who were highly discordant (30% or more). In addition, there was little difference in the magnitude of risk estimates between highly discordant smaller and larger twins. CONCLUSION: After controlling for fetal growth, smaller and larger twins affected by higher levels of birth weight discordance (25% or more) remain at disproportionate risk for neonatal mortality when compared with other smaller or larger twins. Additionally, smaller twins do not have an elevated risk compared with larger twins after adjusting for their different fetal growth distributions. PMID- 12636965 TI - Fetal growth and body proportion in preeclampsia. AB - OBJECTIVE: To evaluate the effects of early- and late-onset preeclampsia on fetal growth and body proportion. METHODS: This was a population-based study based on records of 672,130 pregnancies from the Medical Birth Registry of Norway during 1967-1998. Women with a prior birth, multiple births, those without valid data on the last menstrual period or newborn's crown-heel length, and chronic maternal disease were excluded. RESULTS: In newborns of women with preeclampsia, mean birth weight, crown-heel length, and ponderal index were 4.4%, 0.8%, and 2.6% lower than in births without preeclampsia, respectively. In preterm births, mean differences in birth weight ranged from -11% to -23% against near-equal birth weights in term births. Mean differences in crown-heel length and ponderal index ranged from -1% to -5% and from -5% to -10% before term, respectively. In late preeclampsia, rates of birth weight and crown-heel length above the 90th and 97.5th percentiles and ponderal index above the 97.5th percentile were slightly but significantly higher than in term births without preeclampsia (odds ratios = 1.1-1.5). However, infants born to mothers with preterm preeclampsia were less likely to be heavy, long, or with high ponderal index for gestational age (odds ratios = 0.4-0.6). CONCLUSION: Our results support the hypothesis that preeclampsia is an etiologically heterogeneous disorder that occurs in at least two subsets, one with normal or enhanced placental function, and another involving placental dysfunction and fetal growth restriction, often with asymmetric fetal body proportion, reduced fetal length, and preterm delivery. In future studies, distinguishing between the two subtypes may be important. PMID- 12636966 TI - Antecedents of newborn hearing loss. AB - OBJECTIVE: To determine what factors are associated with newborn hearing loss. METHODS: We retrospectively reviewed hospital records of all neonates found on screening to have hearing loss as well as a cohort with normal audiometric findings from January 1998 through December 2000. The two groups were compared for various maternal, obstetric, and perinatal variables. RESULTS: Sixty-four neonates had hearing loss. The controls consisted of 270 neonates with normal findings. There were no differences between groups when compared for various maternal and perinatal variables. The mean gestational age (35.9 +/- 5.9 weeks versus 37.8 +/- 3.1 weeks, P =.01) and birth weight (2698 +/- 1173 g versus 3195 +/- 816 g, P =.002) were significantly lower in the hearing-loss group. Only 32 of the 64 neonates (50%) had one or more of the ten clinical neonatal high-risk factors for newborn hearing loss. On multivariable analysis, very low birth weight of 1500 g or less (odds ratio [OR] 7; 95% confidence interval [CI] 3, 18; P =.001), 5-minute Apgar scores less than 7 (OR 5; 95% CI 2, 18; P =.009), positive family history of congenital deafness (OR 7; 95% CI 2, 197; P =.02), and structural and chromosomal anomalies (OR 64; 95% CI 14, 292; P =.001) were independently associated with the development of newborn hearing loss. CONCLUSION: Congenital structural and chromosomal anomalies appear to be the most significantly associated risk factors for the development of newborn hearing loss. Very low birth weight, low Apgar scores, and family history are also independently associated with newborn hearing loss. However, most infants with hearing loss have no clinical risk factors. PMID- 12636967 TI - Teaching malpractice litigation in a mock trial setting: a center for perinatal medicine and law. AB - OBJECTIVE: To describe a novel center for perinatal medicine and law in a school of medicine and assess its inaugural presentation of the role of expert witnesses in a mock trial setting. METHODS: The center's first program was an obstetrics and gynecology Grand Rounds that staged an abbreviated mock trial. The case summary was read. An attorney then conducted the direct examination of the plaintiff's obstetrics and gynecology and neonatal expert witnesses. The audience acted as the jury and anonymously voted electronically after the direct examination. The plaintiff's attorney then conducted the cross-examination of the defense experts on issues pertaining to possible bias and expert compensation with only a limited inquiry into substantive medical issues. A second vote was taken. A posttrial panel discussion and questionnaire evaluated the importance of this program to medical education. RESULTS: The first vote indicated 86% of the jury decided that negligent obstetric management contributed substantially to the infant's injury, and 82% of the attendees felt that negligent neonatal care was a substantial factor in the infant's injury. After the cross-examination, 63% of the jurors now felt that negligent obstetric management contributed to the brain damage; whereas only 39% of the participants concluded that negligent newborn care was responsible for the injury. During posttrial discussion, the audience suggested that cross-examination of defense experts on issues of bias rather than medical care negatively affected their perception of the examining attorney's case. CONCLUSION: Analysis of the questionnaire showed that the attendees strongly appreciated experiencing this abbreviated mock trial and indicated that the center was a welcome addition to medical education. PMID- 12636968 TI - Chronic pelvic pain. AB - Chronic pelvic pain is a common and significant disorder of women. It is estimated to have a prevalence of 3.8% in women. Often the etiology of chronic pelvic pain is not clear, as there are many disorders of the reproductive tract, gastrointestinal system, urological organs, musculoskeletal system, and psychoneurological system that may be associated with chronic pelvic pain. The history and physical examination are crucial in evaluating a woman with chronic pelvic pain and must address all of the possible systems potentially involved in chronic pelvic pain, not just the reproductive system. Laboratory and imaging studies should be selectively utilized, as should laparoscopy. Conscious laparoscopic pain mapping has been proposed as a way to improve information derived from laparoscopic evaluations. Treatment of chronic pelvic pain may consist of two approaches. One is to treat chronic pain itself as a diagnosis, and the other is to treat diseases or disorders that might be a cause of or a contributor to chronic pelvic pain. These two approaches are not mutually exclusive, and in many patients effective therapy is best achieved by using both approaches. Treatment of chronic pain as well as treatment of four of the more common disorders associated with chronic pelvic pain (endometriosis, adhesions, irritable bowel syndrome, and interstitial cystitis) are discussed in this review. PMID- 12636969 TI - The problem with evidence-based statistical language. PMID- 12636970 TI - Pelvic umbrella pack for refractory obstetric hemorrhage secondary to posterior uterine rupture. PMID- 12636972 TI - Problems with intrapartum fetal heart rate monitoring interpretation and patient management. PMID- 12636974 TI - Antiphospholipid syndrome in pregnancy: a randomized, controlled trial of treatment. PMID- 12636975 TI - Self-organizing map for cluster analysis of a breast cancer database. AB - The purpose of this study was to identify and characterize clusters in a heterogeneous breast cancer computer-aided diagnosis database. Identification of subgroups within the database could help elucidate clinical trends and facilitate future model building. A self-organizing map (SOM) was used to identify clusters in a large (2258 cases), heterogeneous computer-aided diagnosis database based on mammographic findings (BI-RADS) and patient age. The resulting clusters were then characterized by their prototypes determined using a constraint satisfaction neural network (CSNN). The clusters showed logical separation of clinical subtypes such as architectural distortions, masses, and calcifications. Moreover, the broad categories of masses and calcifications were stratified into several clusters (seven for masses and three for calcifications). The percent of the cases that were malignant was notably different among the clusters (ranging from 6 to 83%). A feed-forward back-propagation artificial neural network (BP-ANN) was used to identify likely benign lesions that may be candidates for follow up rather than biopsy. The performance of the BP-ANN varied considerably across the clusters identified by the SOM. In particular, a cluster (#6) of mass cases (6% malignant) was identified that accounted for 79% of the recommendations for follow up that would have been made by the BP-ANN. A classification rule based on the profile of cluster #6 performed comparably to the BP-ANN, providing approximately 25% specificity at 98% sensitivity. This performance was demonstrated to generalize to a large (2177) set of cases held-out for model validation. PMID- 12636976 TI - Evolutionary computing for knowledge discovery in medical diagnosis. AB - One of the major challenges in medical domain is the extraction of comprehensible knowledge from medical diagnosis data. In this paper, a two-phase hybrid evolutionary classification technique is proposed to extract classification rules that can be used in clinical practice for better understanding and prevention of unwanted medical events. In the first phase, a hybrid evolutionary algorithm (EA) is utilized to confine the search space by evolving a pool of good candidate rules, e.g. genetic programming (GP) is applied to evolve nominal attributes for free structured rules and genetic algorithm (GA) is used to optimize the numeric attributes for concise classification rules without the need of discretization. These candidate rules are then used in the second phase to optimize the order and number of rules in the evolution for forming accurate and comprehensible rule sets. The proposed evolutionary classifier (EvoC) is validated upon hepatitis and breast cancer datasets obtained from the UCI machine-learning repository. Simulation results show that the evolutionary classifier produces comprehensible rules and good classification accuracy for the medical datasets. Results obtained from t-tests further justify its robustness and invariance to random partition of datasets. PMID- 12636977 TI - Building a case-based diet recommendation system without a knowledge engineer. AB - We present a new approach to the effective development of menu construction systems that allow to automatically construct a menu that is strongly tailored to the individual requirements and food preferences of a client. In hospitals and other health care institutions dietitians develop diets for clients which need to change their eating habits. Many clients have special needs in regards to their medical conditions, cultural backgrounds, or special levels of nutrient requirements for better recovery from diseases or surgery, etc. Existing computer support for this task is insufficient-many diets are not specifically tailored for the client's needs or require substantial time of a dietitian to be manually developed. Our approach is based on case-based reasoning, an artificial intelligence technique that finds increasing entry into industrial practice. Our approach goes beyond the traditional case-based reasoning (CBR) approach by allowing an incremental improvement of the system's competency during routine use of the system. The improvement of the system takes place through a direct expert user-system interaction while the expert is accomplishing their tasks of constructing a diet for a given client. Whenever the system performs unsatisfactorily, the expert will need to modify the system-produced diet 'manually', i.e. by entering the desired modifications into the system. Our implemented system, menu construction using an incremental knowledge acquisition system (MIKAS), asks the expert for simple explanations for each of the manual actions he/she takes and incorporates the explanations automatically into its knowledge base (KB) so that the system will perform these manually conducted actions automatically at the next occasion. We present MIKAS and discuss the results of our case study. While still being a prototype, the senior clinical dietitian involved in our evaluation studies judges the approach to have considerable potential to improve the daily routine of hospital dietitians as well as to improve the average quality of the dietary advice given to patients within the limited available time for dietary consultations. Our approach opens up a new avenue towards building highly specialised CBR systems in a more cost effective way. Hence, our approach promises to allow a significantly more widespread development and practical deployment of CBR systems in a large variety of application domains including many medical applications. PMID- 12636978 TI - Decision support tools for clinical trial design. AB - Many published clinical trials are poorly designed, suggesting that the protocol was incomplete, disorganised, or contained errors. This fact motivated the development of a suite of decision support tools for the design of randomised controlled clinical trials. In this paper we describe these tools, discussing both underlying theoretical issues and usage of the tools. The core tool--Design a-Trial (DaT)--critiques data entered so as to guide design of a scientifically and ethically sound trial. DaT outputs a text protocol describing the trial, and a corresponding symbolic representation. Linked to DaT is a tool for authoring plans that form part of the trial. A key feature of this tool is the provision of macros for describing commonly occurring plan constructs. We describe another linked tool which generates solutions to Prolog queries requesting advice on how a plan should be revised so as to comply with safety and efficacy requirements. The user is able to navigate a path through the solution search space by interacting with natural language representations of the Prolog sub-goals. This provides the flexibility to generate useful and informative partial solutions, symbolic and textual, for inclusion in the symbolic plan representation and protocol document, respectively. PMID- 12636979 TI - Characterization of medical time series using fuzzy similarity-based fractal dimensions. AB - This paper attempts to characterize medical time series using fractal dimensions. Existing fractal dimensions like box, information and correlation dimensions characterize the time series by measuring the rate at which the distribution of the time series changes when the length (or radius) of the box (or hypersphere) is changed. However, the measured dimensions significantly vary when the box (or hypersphere) position is changed slightly. It happens because the data points just outside the box (or hypersphere) are not accounted for, and all the data points inside the box or hypersphere are treated equally. To overcome these problems, the hypersphere is converted to a Gaussian, and thus the hard boundary becomes soft. The Gaussian represents the fuzzy similarity between the neighbors and the point around which the Gaussian is constructed. This concept of similarity is exploited to propose a fuzzy similarity-based fractal dimension. The proposed dimension aims to capture the regularity of the time series in terms of how the fuzzy similarity scales up/down when the resolution of the time series is decreased/increased. Experiments on intensive care unit (ICU) data sets show that the proposed dimension characterizes the time series better than the correlation dimension. PMID- 12636981 TI - Antibiotic safety assessment. AB - Antibiotics usually have positive risk-benefit ratios, their adverse effects being generally mild and reversible on treatment cessation. However, severe adverse drug reactions (ADR), associated with significant mortality and morbidity have resulted in the withdrawal of several active antibiotics, including new fluoroquinolones. Adverse reactions to antibiotics are often poorly documented. The purpose of this article is to examine current tools for investigating and preventing antibiotic toxicity and to suggest future lines of investigation. Structure/ADR relationships have been investigated with various antibiotics (beta lactams, macrolides, quinolones, etc.) in an attempt to reduce the risk of adverse reactions. Some reactions can be linked to the drug's stereochemical composition. In the case of quinolones for instance, particularly ofloxacin and its derivatives, experimental data show that individual enantiomers have different toxicities. Another major factor that influences the risk of ADRs in a given population is metabolic variability, due to genetic differences in the relevant drug-metabolizing enzymes. Idiosyncratic antibiotic toxicity can be caused by a chemically reactive metabolite. Recent advances in molecular biology, and especially in individual genomic characterization (DNA chip technology, etc.), could in future be useful for identifying patients who are at a special risk of ADR. Finally, certain pharmacokinetic parameters (AUC, Cmax, etc.) can be used to predict adverse effects. PMID- 12636982 TI - Does short-course antibiotic therapy better meet patient expectations? AB - A pan-European market research study of 3254 patients designed to determine patient attitudes, expectations and behaviour to antibiotic management of mild moderate RTIs, identified three key drivers of perceived antibiotic efficacy: length of antibiotic course, time to onset of symptom relief and time to complete resolution of symptoms. Azithromycin was selected as "drug therapy of shortest dosage schedule" for common outpatient infections. The results demonstrate that once-daily, short-course treatment is perceived to be significantly more effective than longer antibiotic courses and thus, better meets patient expectations of therapy. This perception of efficacy with short-course therapy also correlates with overall satisfaction with management by the physician and compliance with therapy. These findings have important implications for the way physicians manage patients with mild-moderate RTIs. PMID- 12636983 TI - Effect of increasing inoculum sizes of Aspergillus hyphae on MICs and MFCs of antifungal agents by broth microdilution method. AB - In order to investigate the influence of different hyphal inoculum sizes on minimal inhibition concentrations (MICs) and minimum fungicidal concentrations (MFCs) of amphotericin B (AMB), voriconazole and itraconazole, five isolates each of Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger and Aspergillus terreus were studied using a broth microdilution method. Three inoculum sizes were used: 1 x 10(3)-5 x 10(3), 1 x 10(4)-5 x 10(4) and 1 x 10(5)-5 x 10(5) cfu/ml. MICs and MFCs were read at 24 and 48 h at 35 degrees C. For all species tested, AMB MICs and MFCs were minimally affected by inoculum size on. However inoculum size significantly affected MICs and MFCs of voriconazole and itraconazole; there was an increase of up to 6-fold in MICs and MFCs for the various aspergilli when the inoculum increased from 10(3) to 10(5) cfu/ml (P<0.05). Thus azoles showed significant inoculum effects, while AMB showed comparatively minimum inoculum effects against Aspergillus spp. PMID- 12636984 TI - In vitro activity of novel imidazole antifungal agent NND-502 against Malassezia species. AB - The in vitro activity of NND-502, a novel antifungal imidazole compound, was tested against the three major Malassezia species by an agar dilution method with modified Dixon medium and compared with the activities of three reference antifungal drugs of topical use, lanoconazole (LCZ), bifonazole (BFZ) and terbinafine (TBF). The geometric mean (GM)-MICs of NND-502 for 25 strains of M. furfur, 15 strains of M. sympodialis and ten strains of M. slooffiae were approximately 1.4, 0.1 and 1.0 mg/l, respectively, showing the greatest activity against M. sympodialis and the least against M. slooffiae. These values were similar to that of LCZ, but four to 69 times lower than that of BFZ and two to three times lower than that of TBF. The results suggest that NND-502 might be beneficial in the treatment of Malassezia-associated skin diseases. PMID- 12636985 TI - The decline of high drug resistance rate of pulmonary Mycobacterium tuberculosis isolates from a southern Taiwan medical centre, 1996-2000. AB - To investigate the anti-tuberculosis drug resistance pattern of pulmonary tuberculosis isolates in southern Taiwan, we performed a hospital-based surveillance at a southern Taiwan medical centre from 1996 to 2000. The combined drug resistance rates to at least one of four first-line agents (isoniazid, rifampicin, ethambutol, streptomycin) was 52.4%, and to both isoniazid and rifampicin was 11.4%, indicating high resistance rates compared with those reported in the World Health Organization (WHO)/International Union Against Tuberculosis and Lung Disease (IUATLD) global project and in northern Taiwan. The resistance rates to two second-line drugs, cycloserine, and kanamycin, were 75.7 and 23.7%, respectively. A significant decreasing trend in resistance rates to all tested drugs except streptomycin was observed during the 5-year period. The resistance rates in 1996 and 2000 were 43.1 and 16.4% for isoniazid, 23.4 and 9.5% for rifampicin, 23.4 and 12.1% for ethambutol, 92.7 and 50.9% for pyrazinamide. The combined drug resistance rate may not be the most accurate tool as it includes previously treated cases that may inflate the resistance rate and cases without a history of treatment. However, the observation of trends in the susceptibility of pulmonary tuberculosis with the increasing percentages of tuberculosis patients receiving the complete treatment course and the decreasing percentages of cases lost to follow-up in Kaohsiung after the institution of new governmental regulations for case management in 1997, suggest that such intervention programs are useful. PMID- 12636986 TI - Comparison of the susceptibility testing of clinical isolates of Mycobacterium tuberculosis by the XTT colorimetric method and the NCCLS standards method. AB - The susceptibility or resistance of clinical isolates of Mycobacterium tuberculosis were determined by a method incorporating the 2,3-bis(2-methoxy-4 nitro-5-sulphophenyl)-2H-tetrazolium-5-carboxanilide (XTT) and compared with results obtained by the National Committee for Clinical Laboratory Standards approved standard method (M24-T2). One hundred percent of all isolates demonstrated agreement between the susceptibility and resistance to isoniazid, rifampicin, and ethambutol obtained by the two methods, suggesting that the XTT based method could provide a useful means for the rapid determination of antimycobacterial susceptibility of clinical isolates of M. tuberculosis. PMID- 12636987 TI - In vivo susceptibility of Mycobacterium leprae to sitafloxacin (DU-6859a), either singly or in combination with rifampicin analogues. AB - The antimicrobial effects of sitafloxacin (DU-6859a) against Mycobacterium leprae, either singly or in combination with either rifampicin, rifabutin or KRM 1648, were studied using a mouse footpad assay technique and the results were compared with those obtained with ofloxacin. When used singly, the minimum concentrations of sitafloxacin and ofloxacin needed to inhibit completely the growth of M. leprae were 25 and 100 mg per kg body weight per day, respectively, and the effects were bactericidal. Both sitafloxacin and ofloxacin exhibited excellent synergistic effects when combined with either rifabutin or KRM-1648, but not with rifampicin. Thus, incorporation of sitafloxacin and rifabutin (or KRM-1648) in the multidrug regimen for treating leprosy patients is suggested. PMID- 12636988 TI - Impact of sigB mutation on Staphylococcus aureus oxacillin and vancomycin resistance varies with parental background and method of assessment. AB - Previous studies of Staphylococcus aureus transposon insertion mutants showing decreased methicillin or teicoplanin resistance have suggested a role for the RNA polymerase alternative sigma factor SigB in the expression of resistance to these antibiotics. A knockout mutation was created in the S. aureus strain COL sigB gene and its influence on oxacillin and vancomycin resistance was studied in a variety of parental backgrounds. Typically, sigB mutants of methicillin-resistant strains had oxacillin minimum inhibitory concentrations (MICs) one-half of their parent strains. The effect of the sigB mutation appeared to be more dramatic when assessed by population analysis profiles or by growth in liquid culture in shaking flasks than by MIC determinations. Oxacillin MICs of COL and the COLDeltasigB mutant were 400 and 200 mg/l, respectively, by conventional determination and 800 and 100-200 mg/l from population analysis profiles. The COLDeltasigB mutant strain was significantly more inhibited by a range of oxacillin concentrations in a shake flask culture than strain COL. Mutation of sigB caused a decrease in vancomycin resistance in two laboratory derived glycopeptide-intermediate S. aureus strains. The results suggest that some protein products whose expression is controlled by SigB play a role in resistance to cell wall-active antibiotics. PMID- 12636990 TI - Beta-lactamase production in Prevotella and in vitro susceptibilities to selected beta-lactam antibiotics [corrected]. AB - This study looked for beta-lactamase production in 100 Prevotella isolates. MICs were determined for amoxycillin, ticarcillin, amoxycillin+clavulanate, cephalothin, cefuroxime, cefixime, cefpodoxime and cefotaxime using the reference agar dilution method (standard M11 A4, NCCLS). Beta-lactamase activity was detected in 58 of the 100 isolates, 24 of 46 black-pigmented Provotella and 34 of 54 non-pigmented Prevotella. All beta-lactamase-negative strains were susceptible to all beta-lactam antibiotics with the exception of cefuroxime and cefixime. Overall, resistance rates of Prevotella strains were lower for ticarcillin (8%) and celefotaxime (12%) than for the other cephalosporins. All Prevotella isolates were susceptible to amoxycillin and were all inhibited by 2 mg/l or less amoxycillin [corrected]. PMID- 12636989 TI - Comparison of antimicrobial susceptibility between invasive and non-invasive Shigella organisms. AB - Antimicrobial susceptibility was determined for ten strains of Shigella spp. comparing invasive (invasion plasmid containing) and non-invasive members of each strain. The activity of the antimicrobial agents could be classified into three types from the differences between the minimum inhibitory concentration (MIC) for the invasive and non-invasive shigellae. For type 1, there was no difference between the MIC (an MIC gap) for invasive and non-invasive organisms. For type 2, the MIC for the invasive organisms of a strain was higher than that of non invasive organisms of the strain. In the third type, macrolides taken in by shigellae through the type III secretion apparatus, more effectively inhibited the growth of invasive than non-invasive organisms. PMID- 12636991 TI - An approach for the evaluation of synergy between antimicrobials. AB - Synergy can be assessed by a variety of microbiological techniques. Mathematical modelling of synergy data can accomplish similar results and may provide more information than currently available techniques. In this study a combination of 1/4 MIC of aztreonam (AZTR) and 1 MIC of ciprofloxacin (CIPX) showed synergy against Pseudomonas aeruginosa in killing curve studies (based on the two log different criteria) at the end of 24 h. However, re-growth was always observed even when bactericidal concentrations of antibiotics were present. The surviving organisms showed a two-fold increase of MIC to CIPX after the studies and this resistance was not reversible. A mathematical model to quantitatively describe this observation is proposed. PMID- 12636993 TI - Extended-spectrum beta-lactamase types in Klebsiella pneumoniae and Escherichia coli in two Greek hospitals. AB - Seventy-nine Klebsiella pneumoniae and 124 Escherichia coli clinical strains, isolated consecutively during August-October 2001 in two Greek hospitals, were examined for production of extended-spectrum beta-lactamases (ESBLs). Seventy-one (35%) isolates (46 K. pneumoniae and 25 E. coli) were ESBL-positive by phenotypic methods. Isoelectric focusing of beta-lactamases and PCR assays for bla genes showed that SHV-5-type ESBLs were the most frequent (45 isolates, 22%) followed by CTX-M (24 isolates, 12%) and IBC (three isolates, 1.5%). The latter two ESBL types may have been established recently in this setting. PMID- 12636992 TI - In vitro activity of Brazilian medicinal plants, naturally occurring naphthoquinones and their analogues, against methicillin-resistant Staphylococcus aureus. AB - Fourteen extracts from Brazilian traditional medicinal plants used to treat infectious diseases were used to look for potential antimicrobial activity against multiresistant bacteria of medical importance. Staphylococcus aureus strains were susceptible to extracts of Punica granatum and Tabebuia avellanedae. The minimum inhibitory concentrations (MICs) of the total extracts and of additional fractions of these plants were determined by employing strains of methicillin-resistant (MRSA) and -sensitive (MSSA) S. aureus, including isolates of the PFGE clone A, which is prevalent in Brazil and two ATCC reference strains. A mixture of ellagitannins isolated from P. granatum and two naphthoquinones isolated from T. avellanedae demonstrated antibacterial activity against all S. aureus strains tested. Semi-synthetic furanonaphthoquinones (FNQs) showed lower MICs than those exhibited by natural occurring naphthoquinones. The results indicate that these natural products can be effective potential candidates for the development of new strategies to treat MRSA infections. PMID- 12636994 TI - Comparative activity of linezolid, quinupristin-dalfopristin and newer quinolones against Streptococcus pneumoniae. PMID- 12636995 TI - Infection in premature neonates is not associated with higher attributable mortality, but with more neurologic sequellae. PMID- 12636997 TI - Epidemiological situation of rabies in Lithuania from 1990 to 2000. AB - The epidemiological situation of rabies was investigated in Lithuania. Over the last decade, 2277 cases of rabies were registered among wild and domestic animals (mean number of cases per districts was 44). The highest distribution of rabies was found in the districts of Lazdijai and Utena (116 and 81 cases, respectively), and the lowest in the Svencioniai and Zarasai districts (one and eight cases, respectively). It was found that rabies among wildlife comprised 54% with the majority of cases being registered in foxes (626 cases, 27%) followed by raccoon dogs (470 cases, 21%). Within the last 3 years, cases of rabies among foxes and raccoon dogs increased significantly (three and six times, respectively), compared with the period from 1990 to 1997. Among domestic animals, 46% cases of rabies were registered, with cattle comprising 27%, and the cases among cats and dogs at 9 and 8%, respectively. During the period from 1993 to 1997 in Lithuania, 11,385 of humans were attacked by domestic and wild animals, and 21,173 humans were vaccinated against rabies for prophylactic reasons. Our survey has shown that, during the period from 1997 to 2000, the number of people attacked has increased drastically-31,348 (60%), but only 8021 (18%) of them were vaccinated. PMID- 12636998 TI - Antibodies to the CP24 protein of Brucella melitensis lack diagnostic usefulness in ovine brucellosis. AB - The potential diagnostic usefulness of antibodies to the ribosome recycling factor of Brucella melitensis (CP24) was assessed in sheep by an indirect ELISA with purified recombinant CP24. Sera from uninfected animals from the UK (n=44) and from local flocks (n=42), from sheep naturally infected with B. melitensis (n=12) or B. ovis (n=12), and from lambs (n=7) or pregnant ewes (n=6) vaccinated with B. melitensis Rev-1, were assayed. High specific optical densities (OD(with antigen) - OD(without antigen)) were obtained with both the groups of normal sera, which resulted in high cut-off values (1.414 and 1.267, respectively). Only two infected sheep yielded specific OD higher than these cut-off values. No significant difference was found between mean specific OD from B. melitensis- or B. ovis-infected sheep (0.574 and 0.472, respectively), those from vaccinated animals (0.396 and 0.400 for pregnant ewes and lambs, respectively), and those from Brucella-free animals. An inhibition ELISA with soluble CP24 confirmed the specificity of the antibodies detected in normal sera by the indirect ELISA; these antibodies belonged to the IgG class as revealed by the use of a specific conjugate. Sera from infected sheep were all positive for antibodies against lipopolysaccharides and lumazine synthase from Brucella. These results show that anti-CP24 antibodies have no diagnostic role in ovine brucellosis. PMID- 12636999 TI - Epidemiological studies of Brachyspira pilosicoli in two Australian piggeries. AB - The epidemiology of infection with the intestinal spirochaete Brachyspira pilosicoli within pig herds is incompletely understood. To investigate this further, cross-sectional and cohort studies were undertaken on two piggeries. Faeces were subjected to selective culture, and DNA was extracted from growth on the primary media and amplified by polymerase chain reaction (PCR). On one farm, samples from other animal species and the environment were also examined. Isolates were subjected to multilocus enzyme electrophoresis (MLEE) and pulsed field gel electrophoresis (PFGE). The prevalence on farm A (>2000 sows) was 2.4% (95% confidence interval (CI): 0.3, 4.4%). Infection was largely confined to grower/finisher pigs. The six isolates of B. pilosicoli recovered belonged to a single MLEE electrophoretic type (ET) and a single PFGE type. On piggery B, an 80 sow unit located on a research farm, the prevalence amongst growers and finishers was 12.2% (95% CI: 4.7, 19.6%). There was also evidence that weaners were being infected. Ten isolates obtained were genetically heterogeneous, being divided into six ETs and seven PFGE types. One of four isolates in one ET had an identical PFGE type to those on piggery A, and may have been introduced to piggery B in stock from piggery A. On farm B, B. pilosicoli was also detected by PCR in chickens, effluent pond water and wild ducks on the pond. An isolate from the pond belonged to the same ET as one from a pig, whereas the duck isolates were distinct. This study demonstrates the complex epidemiology of B. pilosicoli infections in piggeries. PMID- 12637000 TI - Phenotypic and molecular characterization of bovine Campylobacter fetus strains isolated in Brazil. AB - The objective of the present study was to characterize the phenotypic and molecular aspects of Campylobacter fetus strains isolated from bovine herds with reproductive problems. Thirty-one Brazilian field isolates, together with one reference strain of each subspecies of C. fetus, were analyzed. The strains were submitted to phenotypic identification followed by subspecies characterization using the polymerase chain reaction (PCR) and numeric evaluation of restriction fragment length polymorphism (RFLP) separated by pulsed-field gel electrophoresis (PFGE). Phenotypically, 4 isolates (12.1%) were classified as C. fetus subsp. fetus, and 29 isolates (87.9%) were classified as C. fetus subsp. venerealis. However, according to molecular analysis, only 1 isolate (3.0%) was classified as C. fetus subsp. fetus (the reference strain), whereas 32 isolates (97.0%) were considered C. fetus subsp. venerealis. SalI digestion of C. fetus genomic DNA, obtained from the 33 strains, yielded 7-10 DNA fragments ranging in size from 40 to 373kb, with 12 distinct patterns. Furthermore, the numeric analysis by neighbor-joining of the DNA from the 33 strains resulted in a dendrogram in which 2 distinct groups were identified. It was concluded that phenotypic characterization of C. fetus subspecies might lead to erroneous classification of field isolates. Although RFLP-PFGE is a powerful and reliable technique to characterize C. fetus, it has the inconvenience of being time consuming and laborious. Whereas PCR, besides providing rapid results, was found to be reliable and convenient for the characterization of field isolates of C. fetus. PMID- 12637001 TI - Adherence of Actinobacillus pleuropneumoniae to primary cultures of porcine lung epithelial cells. AB - To study adherence of Actinobacillus pleuropneumoniae to porcine lower respiratory epithelium, a cell culture model was developed using primary cultures of porcine lung epithelial cells (LEC). Adherence assays were performed and results were compared with data obtained with swine kidney cells (SK6). A. pleuropneumoniae efficiently adhered to LEC with up to 62 bacteria per cell after 2h of incubation. Reference strain of serotype 3 (R3) adhered better to LEC than reference strains of serotypes 1 (R1), 7 (R7) and 8 (R8). Overall the adherence to LEC was more rapid and up to 30-fold more efficient than adherence to SK6 cells. In search for the mechanism involved in the adherence event, we tested the effect of LPS which has previously been demonstrated to cause adherence of the pathogen to upper respiratory epithelium. Adherence assays with LPS transposon mutants demonstrated unaltered (mutant with modification in core/lipid A moiety) or even three-fold more adherence (mutants lacking O antigen) compared to the parent micro-organisms. Purified LPS of strains R1, R3, R7 and R8 did not inhibit adherence of R8 to LEC either, suggesting that LPS and particularly the O-antigen are not essential for adherence of A. pleuropneumoniae to LEC. The efficient, LPS independent adherence of A. pleuropneumoniae to LEC cells indicates that A. pleuropneumoniae may carry different, cell type-specific adhesins and that primary cultures of lower respiratory epithelium are valuable infection models in studying A. pleuropneumoniae pathogenesis. PMID- 12637002 TI - Novel protease produced by a Pasteurella trehalosi serotype 10 isolate from a pneumonic bighorn sheep: characteristics and potential relevance to protection. AB - A strain of Pasteurella trehalosi serotype 10, E(CO)-100, isolated from a bighorn sheep that had succumbed to pneumonic pasteurellosis during an epizootic, was compared to well-characterized strains of P. trehalosi serotype 10 and Mannheimia haemolytica serotype 1. The gene for leukotoxin A (lktA) from E(CO)-100 was sequenced and found to be identical on an amino acid basis to a published sequence for lktA from P. trehalosi serotype 10. However, the toxic activity in culture supernatant measured over time for E(CO)-100 was quite different from reference strains. Typically, the ability of the supernatant to lyse target cells increases over time corresponding to the logarithmic growth of the organism, peaks at mid to late phase, then declines gradually. Supernatant from E(CO)-100 exhibited a sharp decline in toxicity after mid-logarithmic growth to undetectable levels. Investigation of this anomaly using a commercial kit with a porcine gelatin/bovine albumin substrate matrix revealed high protease activity in the supernatant of this strain compared to another P. trehalosi serotype 10 and to a M. haemolytica serotype 1. Protease activity was also visualized using gelatin based zymogram gels. This protease was not substrate specific as it was shown to degrade leukotoxin. Activity was neutralized by bighorn sera in a titratable manner. There was an association between the ability to neutralize protease and low pneumonic lung scores in bighorn sheep experimentally challenged with E(CO)-100 (r=0.5, P=0.1). This previously unidentified protease may be an important protective antigen in vaccines designed to prevent pneumonic pasteurellosis resulting from P. trehalosi in bighorn sheep. PMID- 12637004 TI - Comparison of intestinal mucosa homogenate and pure culture of the homologous Lawsonia intracellularis isolate in reproducing proliferative enteropathy in swine. AB - Little information is available on reproduction of proliferative enteropathy (PE) using a virulent pure culture of Lawsonia intracellularis. Reproduction of the disease using PE-diseased mucosa homogenates, however, is well-characterized. The aims of this study were to evaluate and compare clinical signs, growth performance and the severity of lesions in pigs inoculated with intestinal mucosa homogenate or pure culture of the homologous L. intracellularis isolate. Five week-old pigs were inoculated with pure culture of L. intracellularis (isolate PHE/MN1-00; n=10), PE-diseased mucosa (n=10), or control media (n=4). The L. intracellularis isolate PHE/MN1-00 used in the pure culture inoculum was extracted from a fragment of the same intestine used to prepare the mucosa homogenate. Clinical signs and growth performance were evaluated throughout the study. Fecal shedding was evaluated in all animals weekly during the experiment. All animals were euthanized 22 days post-inoculation, the intestines were examined grossly and histologically. Results showed that both the infection procedures reproduced clinical disease, macroscopic and histologic lesions typical of PE. Fecal shedding was detected in animals in both challenge groups. In conclusion, the L. intracellularis isolate PHE/MN1-00 reproduces typical clinical signs and lesions of PE similar to the homologous infection with an intestinal mucosa homogenate. PMID- 12637003 TI - Distribution of Arcobacter species among livestock in Japan. AB - A survey was conducted to examine the distribution of Arcobacter species among livestock in Japan. During May 1999 and May 2000, fecal samples from cattle (n=332) and swine (n=250), chicken cloacal swabs (n=234), and vaginal swabs of cattle (n=61) and swine (n=15) were submitted for the isolation of Arcobacter species. Arcobacter species were isolated from 3.6 and 10.0% of the cattle and swine fecal samples, respectively, along with 14.5% of chicken cloacal swabs. No significant seasonal differences were observed. Species-specific polymerase chain reaction assay showed that A. butzleri was the most prevalent species (83.3, 60.0 and 47.1% of the cattle, swine and chicken isolates, respectively), followed by A. cryaerophilus 1B (16.7, 36.0 and 55.9% of the cattle, swine and chicken isolates, respectively). Of the samples from vaginal swabs, 8.1 and 13.3% were positive for Arcobacter in cattle and swine, respectively. This is the first report demonstrating the distribution of Arcobacter species among livestock in Japan. PMID- 12637005 TI - Colonisation of pet shop puppies with Brachyspira pilosicoli. AB - Anaerobic intestinal spirochaetes of the genus Brachyspira are known to colonise dogs, but relatively little is known about their prevalence, distribution or pathogenic potential. One species, Brachyspira pilosicoli, is thought to cause diarrhoea in dogs, as well as in other animals and humans. To investigate the prevalence and distribution of infection, faecal samples from 49 puppies from six pet shops in the suburbs of Perth, Western Australia were subjected to selective culture for anaerobic intestinal spirochaetes. Growth from the primary plates was also harvested, the DNA extracted and a polymerase chain reaction (PCR) amplification of a portion of the 16S rRNA gene of B. pilosicoli applied. Weakly beta-haemolytic intestinal spirochaetes (WBHIS) grew on plates from 20 of the dogs (40.8%). Seven plates (14.2%) yielded PCR positive amplification for B. pilosicoli. Seven WBHIS isolates were obtained in pure culture, and two of these were shown to be B. pilosicoli by PCR. Application of multilocus enzyme electrophoresis to the seven isolates confirmed that the two PCR positive isolates were B. pilosicoli, whilst the other five belonged to a group previously designated "Brachyspira canis". All the "B. canis" isolates came from healthy puppies, suggesting that this WBHIS is a commensal. Three of the seven puppies with PCR evidence of B. pilosicoli had diarrhoea, but the sample size was small and the association between colonisation and diarrhoea was not statistically significant. Pet shop puppies are commonly infected with intestinal spirochaetes, and may act as a reservoir of B. pilosicoli for other animals and humans. PMID- 12637006 TI - Large-scale protein identification using mass spectrometry. AB - Recent achievements in genomics have created an infrastructure of biological information. The enormous success of genomics promptly induced a subsequent explosion in proteomics technology, the emerging science for systematic study of proteins in complexes, organelles, and cells. Proteomics is developing powerful technologies to identify proteins, to map proteomes in cells, to quantify the differential expression of proteins under different states, and to study aspects of protein-protein interaction. The dynamic nature of protein expression, protein interactions, and protein modifications requires measurement as a function of time and cellular state. These types of studies require many measurements and thus high throughput protein identification is essential. This review will discuss aspects of mass spectrometry with emphasis on methods and applications for large-scale protein identification, a fundamental tool for proteomics. PMID- 12637007 TI - Effect of varying polyglutamate chain length on the structure and stability of ferricytochrome c. AB - The effect of varying polyglutamate chain length on local and global stability of horse heart ferricytochrome c was studied using scanning calorimetry and spectroscopy methods. Spectral data indicate that polyglutamate chain lengths equal or greater than eight monomer units significantly change the apparent pK(a) for the alkaline transition of cytochrome c. The change in pK(a) is comparable to the value when cytochrome c is complexed with cytochrome bc(1). Glutamate and diglutamate do not significantly alter the temperature transition for cleavage of the Met(80)-heme iron bond of cytochrome c. At low ionic strength, polyglutamates consisting of eight or more glutamate monomers increase midpoint of the temperature transition from 57.3+/-0.2 to 66.9+/-0.2 degrees C. On the other hand, the denaturation temperature of cytochrome c decreases from 85.2+/-0.2 to 68.8+/-0.2 degrees C in the presence of polyglutamates with number of glutamate monomers n >or approximately equal 8. The rate constant for cyanide binding to the heme iron of cytochrome c of cytochrome c-polyglutamate complex also decreases by approximately 42.5% with n>or approximately equal 8. The binding constant for the binding of octaglutamate (m.w. approximately 1000) to cyt c was found to be 1.15 x 10(5) M(-1) at pH 8.0 and low ionic strength. The results indicate that the polyglutamate (n>or approximately equal 8) is able to increase the stability of the methionine sulfur-heme iron bond of cytochrome c in spite of structural differences that weaken the overall stability of the cyt c at neutral and slightly alkaline pH. PMID- 12637008 TI - Identification of metabolic enzymes in renal cell carcinoma utilizing PROTEOMEX analyses. AB - PROTEOMEX, an approach which combines conventional proteome analysis with serological screening, is a powerful tool to separate proteins and identify immunogenic components in malignant diseases. By applying this approach, we characterized nine metabolic enzymes which were differentially expressed in renal cell carcinoma (RCC) cell lines and compared their expression profiles to that of normal kidney epithelium cells. Four of these proteins, superoxide dismutase (SODC), triosephosphatase isomerase (TPIS), thioredoxin (THIO) and ubiquitin carboxyl-terminal hydrolase (UBL1) were further analysed for both their constitutive and interferon (IFN)-gamma inducible protein expression pattern in cell lines or tissue specimens derived from RCC or normal kidney epithelium using Western blot analysis and immunohistochemistry, respectively. With the exception of the RCC cell line MZ1940RC, which completely lacks the expression of UBL1, a heterogeneous and variable expression pattern of the different metabolic enzymes was detected in RCC and normal renal epithelium. The highest differences in the expression levels were found for THIO in the RCC cell lines, which was 2-fold upregulated when compared to autologous normal kidney epithelium. Moreover, IFN gamma treatment did not influence the constitutive expression of these metabolic enzymes. Thus, PROTEOMEX represents a valuable approach for the identification of metabolic enzymes which might be used as markers for the diagnosis of RCC. PMID- 12637009 TI - Hydration effects on the protein dynamics in stratum corneum as evaluated by EPR spectroscopy. AB - The uppermost layer of the epidermis, the stratum corneum (SC), was spin-labeled with a sulfhydryl-specific nitroxide reagent to investigate the water content effects upon the protein dynamics directly in the intact tissue. A two-state model for the nitroxide side chain described the coexistence of two spectral components in the electron paramagnetic resonance (EPR) spectra. The so-called strongly immobilized component, S, is associated with the EPR signal of a motionally restricted nitroxide fraction having its N-O group hydrogen bonded to protein (rigid structure) while the weakly immobilized component, W, corresponds to the signal provided by the spin labels with higher mobility (approximately 10 times greater) exposed to the aqueous environment. The relative populations between these two mobility states, S and W, are in thermodynamic equilibrium. The standard Gibbs free energy, enthalpy and entropy changes for transferring the nitroxide side chain from the state contacting the solvent, W, to the one contacting protein, S, indicated that the reduction of the SC water content to below approximately h 0.69 g H(2)O/g dry SC, stabilizes the protein interacting state, S. Upon decreasing the SC hydration level below approximately h 0.69 the segmental motion of the polypeptide chains and the rotational motion of the spin labeled side chain were also constrained. This work can also be useful to improve the spectral analysis of site-directed spin labeling, especially for a more quantitative description in terms of thermodynamic parameters. PMID- 12637011 TI - Equilibrium studies of the effect of difference in sequence homology on the mechanism of denaturation of bovine and horse cytochromes-c. AB - We have carried out equilibrium studies of the effect of the amino acid residue difference in the primary structure of bovine cytochrome-c (b-cyt-c) and horse cyt-c (h-cyt-c) on the mechanism of their folding <--> unfolding processes at pH 6.0 and 25 degrees C. It has been observed that guanidinium chloride (GdmCl) induced denaturation of b-cyt-c follows a two-state mechanism and that of h-cyt-c is not a two-state process. This conclusion is reached from the coincidence and non-coincidence of GdmCl-induced transition curves of bovine and horse proteins, respectively, monitored by measurements of absorbance at 405, 530 and 695 nm and circular dichroism (CD) at 222, 416 and 405 nm. These measurements on h-cyt-c in the presence of GdmCl in the concentration range 0.75-2.0 M also suggest that the protein retains all the native far-UV CD but has slightly perturbed tertiary interaction. The intermediate in the presence of these low denaturant concentrations does not have the structural characteristics of a molten globule as judged by the 8-Anilino-1-napthalene sulfonic acid (ANS) binding and near-UV CD experiments. We have also carried out thermal denaturation studies of bovine and horse cyts-c in the presence of GdmCl monitored by absorbance at 405 nm and far-UV CD at 222 nm. The heat-induced denaturation measurements in the presence of the denaturant show (1) that denaturation of b-cyt-c is a two-state process and that of h-cyt-c does not follow a two-state mechanism, and (2) that the enthalpy change on denaturation of both proteins strongly depends on GdmCl concentration. PMID- 12637010 TI - Postmetamorphic changes in parvalbumin expression in the hindlimb skeletal muscle of the bullfrog, Rana catesbeiana. AB - Anuran amphibians, animals that spend a terrestrial life after metamorphosis, exhibit a marked development of hindlimbs during and after metamorphosis. In order to see whether changes occur in the muscle protein components in the course of postmetamorphic development, we subjected gastrocnemius muscle extracts from growing froglets to two-dimensional electrophoresis (2DE). As a result, we found two proteins to undergo a change in level. One spot, indicating a molecular mass of approximately 12 kDa and an isoelectric point (pI) of 5.0 first became detectable at 45 days after metamorphosis. Another spot, corresponding to a protein of 11 kDa and pI 4.8, was prominent until the former spot appeared. N terminal amino acid sequence analysis and comparison of the spots with those of parvalbumin (PA) revealed that these two proteins were PA alpha and PA beta. Northern blot analysis using PA alpha and PA beta cDNAs as probes revealed that the PA beta mRNA level declined whereas that of PA alpha mRNA rose as the frogs grew. PMID- 12637013 TI - Actin binding of a minispectrin. AB - A "minispectrin" has been constructed from the tail end of the alpha/beta heterodimer, and its actin-binding properties have been characterised. It is a complex of the N-terminal fragment of the beta-subunit consisting of the actin binding domain plus the two first triple-helical repeats beta 1 and beta 2, and the C-terminal fragment of the alpha-subunit containing the repeats alpha 19 and alpha 20 plus the calmodulin-like domain. This minispectrin exists in a dimeric form that contains one copy of each polypeptide and binds to actin in a cooperative manner with an apparent K(d) of 2.5 microM. Calcium seems not to have any effect on its binding to actin. Electron microscopic analysis shows that the minispectrin decorates actin filaments as clusters, and induces formation of actin bundles. This study shows that the actin-binding region of the spectrin alpha/beta heterodimer retains its functional properties in a truncated form and establishes basis for further research on spectrin's structure and function. PMID- 12637012 TI - Tryptophan-tagged cutinase studied by steady state fluorescence for understanding of tag interactions in aqueous two-phase systems. AB - Genetic engineering has been used to construct fusion proteins of Fusarium solani pisi cutinase and tryptophan-based tags, expressed in Saccharomyces cerevisiae, to increase the partitioning in aqueous two-phase systems. The separation systems were composed of thermoseparating polymers (random copolymers of ethylene oxide and propylene oxide, EOPO) and detergents (C(12)EO(n)). In this study, the fluorescence behaviour of the peptide-tagged protein, free peptide tag and tryptophan was investigated. The tryptophan-tagged proteins, cutinase-(WP)(4) and cutinase-TGGSGG-(WP)(4), showed emission spectra similar to the free peptides and tryptophan, indicating solvent exposure of the tag. The influence of polymers and detergents on the fluorescence of tagged proteins was examined. When peptides and tagged proteins were exposed to polymer, a slight blue shift of the emission maximum was observed. Larger blue shifts of the emission maximum were observed when C(12)EO(n) detergents were utilised. The results correlate with aqueous two phase partitioning where addition of C(12)EO(n) detergents results in more extreme partitioning compared to systems containing only polymers. Dynamic light scattering (DLS) measurements of the EOPO copolymers were carried out, showing that the polymers did not aggregate at concentrations used in aqueous two-phase systems. Quenching of fluorescence with iodide for both proteins and peptide tags was studied. Plots according to the Stern-Volmer equation resulted in a linear fit, indicating exposed tryptophan residues for both free peptides and fusion proteins. The quenching constants were similar for both tagged protein and free peptide tag. The fluorescence results indicated that the tryptophan residues in the tag were exposed to the solvent and could interact with detergents and polymers in the two-phase systems. PMID- 12637014 TI - Oligomeric stability of Rapana venosa hemocyanin (RvH) and its structural subunits. AB - The two structural subunits RvH1 and RvH2 were separated after overnight dialysis of Rapana venosa Hc against 130 mM Gly/NaOH buffer, pH 9.6, on an ion exchange column Hiload 26/10 Sepharose Q using a fast performance liquid chromatography (FPLC) system. The reassociation characteristics of these two RvH isoforms and the native molecule were studied in buffers with different pH values and concentrations of Ca(2+) and Mg(2+). Reassociation of mixed RvH subunits was performed over a period of several days using a stabilizing buffer (SB) of pH 7.0 containing different concentrations of Ca(2+) and Mg(2+) ions. After 2 days of dialysis, an RvH subunit mixture of didecamers and multidecamers was observed in the presence of 100 mM CaCl(2) and MgCl(2), though RvH1 and RvH2 are biochemically and immunologically different and have also different dissociation properties. The reassociation, performed at pH 9.6 with 2 mM CaCl(2) and MgCl(2) at 4 degrees C over a period of one to several weeks, led to the formation of decameric oligomers, while didecamers formed predominantly in the SB at pH 7.0. Higher concentrations of calcium and magnesium ions led to a more rapid reassociation of RvH1 resulting in long stable multidecamers and helical tubules, which were stable and slowly dissociated into shorter multidecamers and decamers at higher pH values. The reassociation of the RvH2 structural subunit in the same buffers processed slowly and yielded didecamers, shorter tubule polymers and long multidecamers which are less stable at higher pH values. The stability of RvH isoforms under varying ionic conditions is compared with the stability of keyhole limpet (KLH, Megathura crenulata) hemocyanin (KLH) and Haliotis tuberculata hemocyanin (HtH) isoforms. The process of dissociation and reassociation is connected with changes of the fluorescence intensity at 600 nm, which can be explained by differences in opalescence of the solutions of these two isoforms. The solutions of longer tubule polymers and multidecamers of RvH1 show a higher opalescence compared to the solutions of shorter helical tubules and multidecamers of RvH2. PMID- 12637015 TI - Subdomain organization and catalytic residues of the F factor TraI relaxase domain. AB - TraI from conjugative plasmid F factor is both a "relaxase" that sequence specifically binds and cleaves single-stranded DNA (ssDNA) and a helicase that unwinds the plasmid during transfer. Using limited proteolysis of a TraI fragment, we generated a 36-kDa fragment (TraI36) retaining TraI ssDNA binding specificity and relaxase activity but lacking the ssDNA-dependent ATPase activity of the helicase. Further proteolytic digestion of TraI36 generates stable N terminal 26-kDa (TraI26) and C-terminal 7-kDa fragments. Both TraI36 and TraI26 are stably folded and unfold in a highly cooperative manner, but TraI26 lacks affinity for ssDNA. Mutational analysis of TraI36 indicates that N-terminal residues Tyr(16) and Tyr(17) are required for efficient ssDNA cleavage but not for high-affinity ssDNA binding. Although the TraI36 N-terminus provides the relaxase catalytic residues, both N- and C-terminal structural domains participate in binding, suggesting that both domains combine to form the TraI relaxase active site. PMID- 12637016 TI - Isolation and characterization of denatured serum albumin from rats with endotoxicosis. AB - Due to its rapid breakdown in the body, denatured serum albumin has not been identified in biological samples. In this study we attempted to determine whether denatured albumin could be identified in rats with endotoxicosis. Male Wistar rats were injected with lipopolysaccharide (LPS; 5 mg/kg body weight). Plasma albumin concentration decreased to one-third the normal level at 2 days after the injection. By using the purified IgG against the specific epitope of chemically denatured albumin, two immunoreactive plasma proteins (bands D2 and D3) were identified by native PAGE followed by Western blot analysis. The plasma concentration of these two proteins increased significantly at 1 and 1.5 days after LPS injection. Peptide mass fingerprinting using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI/TOF-MS) identified these two proteins as serum albumin. In order to characterize their conformational nature, ion-exchange chromatography was used to isolate D2 and D3 albumins from rats injected with LPS. Far- and near-UV circular dichroism (CD), tryptophan and 1-anilino-8-naphthalenesulfonate (ANS) fluorescence, and proteolytic susceptibility showed conformational alterations in the D2 and D3 albumins as compared with native albumin. These data indicate the presence of denatured albumin in circulating rat plasma, and this fact may contribute to a further understanding of the molecular mechanisms of albumin breakdown in physiological and pathophysiological conditions. PMID- 12637017 TI - Kinetic mechanism of choline oxidase from Arthrobacter globiformis. AB - Choline oxidase catalyzes the four-electron oxidation of choline to glycine betaine, with betaine-aldehyde as intermediate and molecular oxygen as primary electron acceptor. The enzyme is capable of accepting betaine-aldehyde as a substrate, allowing the investigation of the reaction mechanism for both the conversion of choline to the aldehyde intermediate and of betaine-aldehyde to glycine-betaine. The steady state kinetic mechanism has been determined at pH 7 with choline and betaine-aldehyde as substrate to be sequential, consistent with oxygen reacting with the reduced enzyme before release of betaine-aldehyde or glycine-betaine, respectively. A K(m) value < or =20 microM has been estimated for betaine-aldehyde based on the kinetic pattern with a y-intercept seen in a plot of 1/rate versus 1/[oxygen]. The kinetic data suggest that betaine-aldehyde predominantly remains bound at the active site during turnover of the enzyme with choline. In agreement with such a conclusion, less than 10% betaine-aldehyde has been found in the reaction mixture under enzymatic turnover with saturating concentrations of choline. The k(cat) values were 6.4+/-0.3 and 15.3+/-2.5 s(-1) for choline and betaine-aldehyde, respectively, suggesting that a kinetic step in the oxidation of choline to the aldehyde intermediate must be partially rate limiting for catalysis. Cleavage of the CH bond of choline as being partially rate-limiting for catalysis is discussed. PMID- 12637019 TI - Binding of antibromelain monomeric Fab' improves the stability of stem bromelain against inactivation. AB - Antienzyme polyclonal antibodies against stem bromelain were raised in male albino rabbits and the Fab' monomers isolated from the IgG of the immune sera. Incubation of bromelain with the Fab' resulted in binding and gel filtration of the resulting complex suggested a 1:1 stoichiometry. Complexing with the Fab' resulted in significant stabilization of bromelain against thermal inactivation and alkaline pH. PMID- 12637018 TI - Different degree of cooperativity in adult, embryonic and mutated mouse muscle nicotinic receptors. AB - Adult and embryonic nicotinic receptors expressed in COS cells have similar affinities for acetylcholine but differ in their Hill coefficient. Parameters of wild-type receptors were compared with those of receptors with mutated delta and gamma subunits in selected negatively charged amino acids, which were expected to participate in agonist binding. A tentative scheme of affinities, allosteric interactions and channel gating efficacy was used for assessing the role of mutated amino acids in the channel function. In three models, the parameters of wild-type embryonic and adult receptors were compared with those of receptors with mutated delta and gamma subunits. The analysis of different models of channel activation indicates that negatively charged amino acids which were mutated in the delta subunit in embryonic receptors participate in channel gating and in allosteric interactions between subunits rather than directly in agonist binding. Changes in the gamma subunit in the embryonic receptors and delta subunit in the adult receptors could equally affect agonist binding, allosteric coupling between subunits or channel gating. PMID- 12637020 TI - Chimeric sensory kinases containing O2 sensor domain of FixL and histidine kinase domain from thermophile. AB - To explore the functional mechanism of inter-domain interaction in a sensor histidine kinase, five chimeric sensory kinases were constructed. In each of these chimeric proteins (CskA254, CskA264, CskA274, CskA284, and CskA294), the sensor domain of heme-based O(2) sensor FixL, obtained from Sinorhizobium meliloti, was fused with the histidine kinase domain from a hyperthermophile, Thermotoga maritima, each at a systematically different position. The UV-visible (UV-vis), resonance Raman (RR), and circular dichroism (CD) spectral characteristics of the CskAs indicated that the secondary and heme environmental structures of all five CskAs examined are identical to those of FixL. In spite of these structural similarities, all CskAs did not exhibit O(2)-dependent regulation of autophosphorylation activity. Furthermore, their functional properties were much different from those of FixL: The O(2) binding affinity and the autophosphorylation activity for CskA254, CskA264, and CskA274 were similar to those of the truncated sensor and histidine kinase domain, whereas CskA284 and CskA294 display extremely low O(2) affinity and low autophosphorylation activity, as compared with each truncated domain. These observations indicated that the interdomain interaction was presented in those CskAs, and that interaction could be related to the O(2)-dependent regulatory interaction of FixL. In the present study, we demonstrated that the interaction in the physiological sensor histidine kinase would be strictly and finely controlled to mediate the signal ligation dependent autophosphorylation activity in its histidine kinase domain. PMID- 12637021 TI - The atypical lipase B from Candida antarctica is better adapted for organic media than the typical lipase from Thermomyces lanuginosa. AB - Candida antarctica lipase B (CALB) and Thermomyces lanuginosa lipase (TLL) were evaluated as catalysts in different reaction media using hydrolysis of tributyrin as model reaction. In o/w emulsions, the enzymes were used in the free form and for use in monophasic organic media, the lipases were adsorbed on porous polypropylene (Accurel EP-100). In monophasic organic media, the highest specific activity of both lipases was obtained in pure tributyrin at a water activity of >0.5 and at an enzyme loading of 10 mg/g support. With tributyrin emulsified in water, the specific activities were 2780 micromol min(-1) mg(-1) for TLL and 535 micromol min(-1) mg(-1) for CALB. Under optimal conditions in pure tributyrin, CALB expressed 49% of the activity in emulsion (264 micromol min(-1) mg(-1)) while TLL expressed only 9.2% (256 micromol min(-1) mg(-1)) of its activity in emulsion. This large decrease is probably due to the structure of TLL, which is a typical lipase with a large lid domain. Conversion between open and closed conformers of TLL involves large internal movements and catalysis probably requires more protein mobility in TLL than in CALB, which does not have a typical lid region. Furthermore, TLL lost more activity than CALB when the water activity was reduced below 0.5, which could be due to further reduction in protein mobility. PMID- 12637022 TI - Functional consequences of deleting the two C-terminal residues of the scorpion toxin BmTX3. AB - We deleted the two C-terminal residues of the scorpion toxin BmTx3, a peptidyl inhibitor of a transient A-type K(+) current in striatum neurons in culture, to assess their contribution to receptor recognition. The sBmTX3-delYP analog was shown to have a native-like structure in one-dimensional 1H-nuclear magnetic resonance (NMR) spectroscopy. We found that sBmTX3-delYP bound to its receptor less efficiently than the wild-type molecule (by a factor of about 10(5)) in binding assays with rat brain membranes, and that this molecule did not block the A-type K(+) current (at a concentration of 35 microM) in whole-cell patch clamp experiments with striatum neurons. Also, these results show that the A-type K(+) channel blocked by BmTX3 should have a canonical K(+) channel pore structure. PMID- 12637023 TI - Podophyllum peltatum possesses a beta-glucosidase with high substrate specificity for the aryltetralin lignan podophyllotoxin. AB - A beta-glucosidase with high specificity for podophyllotoxin-4-O-beta-D glucopyranoside was purified from the leaves of Podophyllum peltatum. The 65-kDa polypeptide had optimum activity at pH 5.0 and was essentially inactive at pH 6.5 or above. Maximum catalytic activity of this glucosidase was obtained at 45 degrees C, but the enzyme was not heat stable. This beta-glucosidase displayed higher substrate specificity for podophyllotoxin-4-O-beta-D-glucopyranoside than for the other lignans tested, and for the (1-->3) linkage of laminaribiose than for other glucosidic linkages. PMID- 12637024 TI - Deletion of N-terminal amino acids from human lecithin:cholesterol acyltransferase differentially affects enzyme activity toward alpha- and beta substrate lipoproteins. AB - Lecithin:cholesterol acyltransferase (LCAT) is the enzyme responsible for generation of the majority of the cholesteryl esters (CE) in human plasma. Although most plasma cholesterol esterification occurs on high-density lipoprotein (HDL), via alpha-LCAT activity, esterification also occurs on low density lipoprotein (LDL) via the beta-activity of the enzyme. Computer threading techniques have provided a three-dimensional model for use in the structure function analysis of the core and catalytic site of the LCAT protein, but the model does not extend to the N-terminal region of the enzyme, which may mediate LCAT interaction with lipoprotein substrates. In the present study, we have examined the functional consequences of deletion of the highly conserved hydrophobic N-terminal amino acids (residues 1-5) of human LCAT. Western blot analysis showed that the mutant proteins (Delta 1-Delta 5) were synthesized and secreted from transfected COS-7 cells at levels approximately equivalent to those of wild-type hLCAT. The secreted proteins had apparent molecular weights of 67 kDa, indicating that they were correctly processed and glycosylated during cellular transit. However, deletion of the first residue of the mature LCAT protein (Delta 1 mutant) resulted in a dramatic loss of alpha-LCAT activity (5% of wild type using reconstituted HDL substrate, rHDL), although this mutant retained full beta-LCAT activity (108% of wild-type using human LDL substrate). Removal of residues 1 and 2 (Delta 2 mutant) abolished alpha-LCAT activity and reduced beta-LCAT activity to 12% of wild type. Nevertheless, LCAT Delta 1 and Delta 2 mutants retained their ability to bind to rHDL and LDL lipoprotein substrates. The dramatic loss of enzyme activity suggests that the N-terminal residues of LCAT may be involved in maintaining the conformation of the lid domain and influence activation by the alpha-LCAT cofactor apoA-I (in Delta 1) and/or loss of enzyme activity (in Delta 1-Delta 5). Since the Delta 1 and Delta 2 mutants retain their ability to bind substrate, other factor(s), such as decreased access to the substrate binding pocket, may be responsible for the loss of enzyme activity. PMID- 12637025 TI - Identification and characterization of a new ligand-binding site in FnbB, a fibronectin-binding adhesin from Streptococcus dysgalactiae. AB - Streptococcus dysgalactiae S2, a bovine mastitis isolate, expresses the fibronectin (Fn)-binding adhesin FnbB. Here, we describe a new fibronectin binding domain called UFnBD, located 100 amino acid N-terminal to the primary repetitive Fn-binding domain (FnBRD-B) of FnbB. UFnBD interacted with N-terminal region of Fn (N29) and this binding was mostly mediated by type I module pair 2-3 of N29 fragment, whereas FnBRD-B mainly bound to type I module pair 4-5. Furthermore, UFnBD inhibited adherence of S. dysgalactiae to Fn but at lower level as compared to FnBRD-B. UFnBD exclusively shared antigenic properties with the Fn-binding unit Du of FnbpA from Staphylococcus aureus but not with ligand binding domains or motifs of other adhesins, while Fn-induced determinants of FnBRD-B and other adhesins appeared to be conformationally related. Consistent with this, a monoclonal antibody 7E11 generated from a mouse immunized with FnbB, and that recognized UFnBD did not cross-react with FnBRD-B. The epitope for 7E11 was mapped to 40 amino acid long segment within UFnBD and interaction between the antibody and the epitope was specifically induced by Fn or N29. A similar antibody epitope was observed in Streptococcus pyogenes strains suggesting the presence of an adhesin bearing epitope related to FnbB. PMID- 12637026 TI - Kinetic and mechanistic analysis of the association and dissociation of inhibitors interacting with secreted aspartic acid proteases 1 and 2 from Candida albicans. AB - In order to elucidate the characteristics of different aspartic proteases (Sap) secreted by Candida albicans, the kinetics of the interaction (k(on), k(off)) between Sap1 and Sap2 with acetyl-pepstatin and pepstatin A was determined at different pH by biosensor technology. The enzymes were biotinylated and coupled to a streptavidin-coated sensor chip, whereupon acetyl-pepstatin or pepstatin A was injected and the interaction was measured in real time. Sap2 showed a faster k(on) and a higher affinity for acetyl-pepstatin than Sap1, regardless of pH. The values for both k(on) and k(off) decreased with increased pH from 3.8 to 5.0, except for the k(off) for Sap1, which was only influenced by the pH change from 3.8 to 4.4. Binding of acetyl-pepstatin to Sap1 or Sap2 obviously proceeds by a different mechanism than dissociation of the inhibitor. Association appears to be coupled to protonation of a catalytic aspartic acid residue, consistent with reduced k(on) values at higher pH. In contrast, the stability of the complex is reduced at lower pH due to reduced hydrogen bonding capacity of aspartic acid residues acting as hydrogen bond acceptors. Differences in the number and distribution of charged nonactive site residues in Sap1 and Sap2 evidently result in different electrostatic properties of the binding sites, primarily influencing the association step. PMID- 12637027 TI - CTP:glycerol 3-phosphate cytidylyltransferase (TarD) from Staphylococcus aureus catalyzes the cytidylyl transfer via an ordered Bi-Bi reaction mechanism with micromolar K(m) values. AB - CTP:glycerol 3-phosphate cytidylyltransferase catalyzes the formation of CDP glycerol, an activated form of glycerol 3-phosphate and key precursor to wall teichoic acid biogenesis in Gram-positive bacteria. There is high sequence identity (69%) between the CTP:glycerol 3-phosphate cytidylyltransferases from Bacillus subtilis 168 (TagD) and Staphylococcus aureus (TarD). The B. subtilis TagD protein was shown to catalyze cytidylyltransferase via a random mechanism with millimolar K(m) values for both CTP and glycerol 3-phosphate [J. Biol. Chem. 268, (1993) 16648] and exhibited negative cooperativity in the binding of substrates but not in catalysis [J. Biol. Chem. 276, (2001) 37922]. In the work described here on the S. aureus TarD protein, we have elucidated a steady state kinetic mechanism that is markedly different from that determined for B. subtilis TagD. Steady state kinetic experiments with recombinant, purified TarD employed a high-performance liquid chromatography assay developed in this work. The data were consistent with a ternary complex model. The K(m) values for CTP and glycerol 3-phosphate were 36 and 21 microM, respectively, and the k(cat) was 2.6 s(-1). Steady state kinetic analysis of the reverse (pyrophosphorylase) reaction was also consistent with a ternary complex model. Product inhibition studies indicated an ordered Bi-Bi reaction mechanism where glycerol 3-phosphate was the leading substrate and the release of CDP-glycerol preceded that of pyrophosphate. Finally, we investigated the capacity of S. aureus tarD to substitute for tagD in B. subtilis. The tarD gene was placed under control of the xylose promoter in a B. subtilis 168 mutant defective in tagD (temperature-sensitive, tag-12). Growth of the resulting strain at the restrictive temperature (47 degrees C) was shown to be xylose-dependent. PMID- 12637028 TI - Hipposin, a histone-derived antimicrobial peptide in Atlantic halibut (Hippoglossus hippoglossus L.). AB - A novel 51-residue antimicrobial peptide (AMP) from the skin mucus of Atlantic halibut (Hippoglossus hippoglossus L.) was isolated using acid extraction, and cationic exchange and reversed phase chromatography. The complete amino acid sequence of the AMP, termed hipposin, was determined by automated Edman degradation and mass spectrometry to be SGRGKTGGKARAKAKTRSSRAGLQFPVGRVHRLLRKGNYAHRVGAGAPVYL. The N-terminal amino group was acetylated. The theoretical mass of hipposin was calculated to be 5458.4 Da, which was in good agreement with the mass of 5459 Da determined by matrix assisted laser desorption ionization mass spectrometry (MALDI-MS). Hipposin was shown to be derived from histone H2A by PCR amplifying the encoding sequences from Atlantic halibut genomic DNA. The peptide showed sequence similarity with the 39-mer AMP buforin I of Asian toad and the 19-mer AMP parasin I of catfish. Fifty of the fifty-one residues in hipposin were identical to the N-terminal region of histone H2A from rainbow trout. Hipposin showed strong antimicrobial activity against several Gram-positive and Gram-negative bacteria and activity could be detected down to hipposin concentrations of 0.3 microM (1.6 microg/ml). Hipposin without N-terminal acetylation was prepared by solid-phase peptide synthesis and shown to have the same antimicrobial activity as the natural acetylated peptide. Thus, hipposin is a new broad-spectrum histone-derived AMP found in the skin mucus of Atlantic halibut. PMID- 12637029 TI - Characterization of an ectonucleoside triphosphate diphosphohydrolase 1 activity in alkaline phosphatase-depleted rat osseous plate membranes: possible functional involvement in the calcification process. AB - An ectonucleoside triphosphate diphosphohydrolase 1 (NTPDase1) activity present in alkaline phosphatase-depleted rat osseous plate membranes, obtained 14 days after implantation of demineralized bone particles in the subcutaneous tissue of Wistar rats, was characterized. At pH 7.5, NTPDase1 hydrolyzed nucleotide triphosphates at rates 2.4-fold higher than those of nucleotide diphosphates, while the hydrolysis of nucleotide monophosphates and non-nucleotide phosphates was negligible. NTPDase 1 hydrolyzed ATP and ADP following Michaelis-Menten kinetics with V=1278.7+/-38.4 nmol Pi/min/mg and K(M)=83.3+/-2.5 microM and V=473.9+/-18.9 nmol Pi/min/mg and K(M)=150.6+/-6.0 microM, respectively, but in the absence of magnesium and calcium ions, ATP or ADP hydrolysis was negligible. The stimulation of the NTPDase1 by calcium (V=1084.7+/-32.5 nmol Pi/min/mg; and K(M)=377.8+/-11.3 microM) and magnesium (V=1367.2+/-41.0 nmol Pi/min/mg and K(M)=595.3+/-17.8 microM) ions suggested that each ion could replace the other during the catalytic cycle of the enzyme. Oligomycin, ouabain, bafilomycin A(1), theophylline, thapsigargin, ethacrynic acid, P(1),P(5)-(adenosine-5') pentaphosphate and omeprazole had negligible effects on the hydrolysis of ATP and ADP by NTPDase1. However, suramin and sodium azide were effective inhibitors of ATP and ADP hydrolysis. To our knowledge this is the first report suggesting the presence of NTPDase1 in rat osseous plate membranes. Considering that the ectonucleoside triphosphate diphosphohydrolase family of enzymes participates in many regulatory functions, such as response to hormones, growth control, and cell differentiation, the present observations raise interesting questions about the participation of this activity in the calcification process. PMID- 12637030 TI - Cardiac dynamics in corticotropin-releasing factor receptor subtype-2 deficient mice. AB - The action of corticotropin-releasing factor (CRF) is mediated by two recently identified receptors, CRFR1 and CRFR2, that differ with respect to their anatomical distribution and pharmacologic ligand-binding properties. Here we show by an analysis of circadian heartbeat interval fluctuations that CRFR2-deficiency in mice does not interfere with the dynamical mechanisms underlying the control of heart rate. Hence, intact CRFR2 would not constitute an indispensable requirement of physiologic cardiac rhythm regulation. However, both CRFR2 knockout (-/-) and wildtype control (+/+) mice showed altered dynamical properties of cardiac interbeat fluctuations in contrast to homogenetic inbred strains of mice (C57BL/6N and C57BL/6J). The results stress the impact of genetic background and support the generalized notion that transgenic 129/Sv-derived knockout mice exhibit altered cardiac dynamics which is interpreted to reflect an attenuation of neuroautonomic sympatho-vagal antagonism. PMID- 12637031 TI - Effects of haloperidol and clozapine on neuropeptide Y-like immunoreactivity in the nucleus accumbens and striatum of rats pretreated with psychostimulants. AB - It has been shown that the synthesis, release and levels of neuropeptide Y (NPY), a peptide linked to the dopamine system, are altered by stimulants with psychotomimetic properties and by antipsychotic drugs. This study was designed to evaluate the effect of 3-day haloperidol (HAL) (2 mg/kg i.p.) or clozapine (CLOZ) (25 mg/kg i.p.) treatment on neuropeptide Y-like immunoreactivity (NPY-LI) in nucleus accumbens and striatum (caudate-putamen) in rats pretreated with d amphetamine or phencyclidine. D-amphetamine (5 mg/kg s.c. twice daily for 6 days and once on day 7) and phencyclidine (10 mg/kg i.p. once daily for 2 days and 15 mg/kg once on day 3) induced marked stereotypy with different symptomatology. Stereotypy is thought to resemble psychosis-related behavior. The first dose of either HAL or CLOZ was given 3 or 2h after the final d-amphetamine or phencyclidine injection, respectively. The control groups were injected with either saline alone, saline instead of psychostimulants, which was followed by antipsychotics, or psychostimulants followed by saline. Rats were sacrificed 24h after antipsychotics or 72 h after the last psychostimulant dose. Both psychostimulants similarly reduced nucleus accumbens and striatal NPY-LI. In saline-pretreated rats, HAL and CLOZ decreased nucleus accumbens NPY-LI, but only HAL decreased striatal NPY-LI. In both the structures examined the effects of HAL and d-amphetamine on NPY-LI were additive. HAL slightly enhanced but CLOZ reversed the phencyclidine-induced decrease in accumbens NPY-LI. The present study has shown that HAL and CLOZ produce different effects on nucleus accumbens and striatal NPY-LI in d-amphetamine or phencyclidine pretreated rats, and it suggests that these effects are related particularly to the dopaminergic and glutamatergic systems whose activities were altered by psychostimulants. PMID- 12637032 TI - CCK2 receptor-deficient mice have increased sensitivity of dopamine D2 receptors. AB - The present study supports a role of CCK(2) receptors in the regulation of dopamine neurones. In pharmacological studies conducted on male CCK(2) receptor deficient mice the changes in the activity of dopamine system were established. A low dose of dopamine agonist apomorphine (0.1 mg/kg), stimulating the pre synaptic dopamine receptors, induced significantly stronger suppression of locomotor activity in mutant mice (-/-) compared to their wild-type littermates (+/+). The administration of amphetamine (3-6 mg/kg), a drug increasing dopamine release, caused a dose-dependent stimulation of locomotor activity in wild-type mice. In mice lacking CCK(2) receptors, a lower dose of amphetamine (3 mg/kg) tended to suppress the motor activity, whereas the higher dose (6 mg/kg) induced the significantly stronger motor stimulation in mutant mice. Moreover, in the CCK(2) receptor-deficient mice the affinity of dopamine D(2) receptors, but not 5 HT(2) receptors, was increased. Altogether, the targeted genetic suppression of CCK(2) receptors increased the sensitivity of pre- and post-synaptic dopamine D(2) receptors. PMID- 12637033 TI - The role of the amino-terminal domain of tachykinins in neurokinin-1 receptor signaling and desensitization. AB - Neurokinin A (NKA) has previously been shown to be a full agonist of the neurokinin-1 receptor (NK-1R) but is only able to cause partial homologous desensitization of the receptor compared to substance P (SP). NKA and SP share the same amino acid sequence at their C-terminal active site domains but differ in structure at their N-terminal domains. These observations have led to the proposal that the N-terminal domains of tachykinin peptides affect the desensitization but not the agonist activities of the peptides. Some of the preprotachykinin proteins contain SP and the NKA-like tachykinins neuropeptide K (NPK) and neuropeptide gamma (NPgamma), which contain NKA at their C-terminals and are N-terminally extended. In this study, the abilities of NKA, NPK, and NPgamma to stimulate NK-1R second messenger (IP(3)) signaling and rapid homologous desensitization of the NK-1R were examined. In addition, a similar analysis was performed using several nonmammalian tachykinin peptides in order to obtain additional insight into the role of the tachykinin N-terminal domain in these NK-1R functions. NPK and NPgamma were found, like NKA, to be full agonists of rat NK-1R IP(3) signaling but, unlike NKA, were also able to cause full rapid homologous desensitization of the receptor. The extended N-terminal domains of NPK and NPgamma thus increase the desensitization activities of these NKA-like peptides. Of the nonmammalian tachykinins tested, all were full agonists but kassinin and eledoisin had only partial homologous desensitization activity, suggesting that the N-terminal structures of these peptides also differentially affect agonist versus desensitization activities of the NK-1R. PMID- 12637034 TI - Kinin B1 receptor antagonists inhibit diabetes-induced hyperalgesia in mice. AB - Insulin-dependent diabetes mellitus (type 1 diabetes) is an inflammatory autoimmune disease associated with vascular permeability changes leading to many complications including nephropathy, retinopathy, neuropathy, hypertension and hyperalgesia. The bradykinin B(1) receptors (BKB(1)-R) were recently found to be upregulated alongside the development of type 1 diabetes and to be involved in its complications. Kinins are important mediators of a variety of biological effects including cardiovascular homeostasis, inflammation and nociception. In the present study, we studied the effect of a selective BKB(1)-R agonist desArg(9)-BK (DBK) and two selective receptor antagonists, the R-715 (Ac-Lys-[D beta Nal(7), Ile(8)] desArg(9)-BK) and the R-954 (Ac-Orn-[Oic(2), alphaMe Phe(5), D-beta Nal(7), Ile(8)] desArg(9)-BK) on diabetic hyperalgesia. Type 1 diabetes was induced in male CD-1 mice via a single injection of streptozotocin (STZ, 200mg/kg, i.p.), one week before the test. Nociception, a measure of hyperalgesia, was assessed using the plantar stimulation (Hargreaves) and the tail-immersion tests. The induction of type 1 diabetes provoked a significant hyperalgesic activity in diabetic mice, causing an 11% decrease in plantar stimulation reaction time and 13% decrease in tail-immersion reaction time, compared to normal mice. Following acute administration of R-715 (100-600 microg/kg, i.p.), or R-954 (50-400 microg/kg, i.p.), the STZ-induced hyperalgesic activity was blocked in a dose-dependent manner and the hot plate and tail immersion latencies of diabetic mice returned to normal values observed in control healthy mice. In addition, the acute administration of DBK (400 microg/kg, i.p.) significantly potentiated diabetes-induced hyperalgesia, an effect that was totally reversed by R-715 (1.6-2.4 mg/kg, i.p.) and R-954 (0.8 1.2mg/kg, i.p.). These results provide further evidence for the implication of the BKB(1)-R in type 1 diabetic hyperalgesia and suggest a novel approach in the treatment of this complication using the BKB(1)-R antagonists. PMID- 12637035 TI - Effects of glucagon-like peptide-1 (7-36) amide on neurohypophysial hormone secretion induced by acute hyperosmotic challenge. AB - This study was designed to investigate possible effects of glucagon-like peptide 1 (7-36) amide on the vasopressin and oxytocin release induced by acute peripheral or central osmotic stimulation. In the first series of experiments, rats were injected intraperitoneally with the isotonic (0.15 M) or hypertonic (1.5 M) NaCl solution and then, intracerebroventricularly, with either 1 microg glucagon-like peptide-1 (7-36) amide dissolved in 5 microl of isotonic saline or with the vehicle only. In the second study, 1 microg glucagon-like peptide-1 (7 36) amide, dissolved in isotonic or hypertonic (0.6 M) saline, was injected into the cerebroventricular system. Control rats were treated with isotonic or hypertonic saline only. All the animals were decapitated 10 min after the intracerebroventricular injection. Glucagon-like peptide-1 (7-36) amide enhanced significantly the basal secretion of vasopressin and oxytocin. Moreover, this peptide increased additionally the release of both neurohypophysial hormones stimulated previously by peripheral osmotic challenge. On the other hand, the peptide increased the oxytocin but not vasopressin secretion brought about by an intracerebroventricular injection of hypertonic saline thus suggesting that the central osmotic stimulation decreases the sensitivity of vasopressin neurons to glucagon-like peptide-1 (7-36) amide. It is concluded that glucagon-like peptide 1 (7-36) amide may affect the secretory activity of the hypothalamo neurohypophysial system under acute osmotic challenge. PMID- 12637036 TI - Electroconvulsive stimuli alter nerve growth factor but not brain-derived neurotrophic factor concentrations in brains of a rat model of depression. AB - Nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are proteins involved in neuronal survival and plasticity of dopaminergic, cholinergic and serotonergic neurons in the central nervous system (CNS). Moreover, it has been hypothesized that these molecules play a role in the pathophysiology as well as treatment of depression. Using an animal model of depression, the Flinders Sensitive Line (FSL) rats and their controls, the Flinders Resistant Line (FRL), we investigated the effects of electroconvulsive stimuli (ECS) on brain NGF and BDNF. ECS or SHAM ECS were administered eight times, with a 48-h interval between each treatment. NGF and BDNF were measured with enzyme-linked immunosorbent assay (ELISA). In the hippocampus ECS increased NGF concentration in FSL but not FRL rats. ECS decreased NGF concentration in the frontal cortex of FSL rats. In both FSL and FRL rats ECS increased NGF levels in the striatum. In contrast, ECS did not change BDNF concentration in hippocampus, frontal cortex and striatum of FSL and FRL rats. Our data support the notion that neurotrophin concentrations may be altered by ECS. PMID- 12637037 TI - Not only vasopressin, but also the intracellular messenger protein kinase Calpha in the suprachiasmatic nucleus correlates with expression of circadian rhythmicity in voles. AB - The suprachiasmatic nucleus (SCN) is the locus of the main pacemaker for circadian behavioral rhythms. In common voles, variation in circadian behavioral rhythmicity correlates with vasopressin (AVP) immunoreactive cells in the SCN. Here we studied the immunostaining of four AVP linked Ca(2+)-dependent protein kinase C (PKC) isoforms (PKCalpha, PKCbeta1, PKCbeta2, and PKCgamma) at the beginning of the light period, and conclude that PKCalpha is highly expressed in the vole SCN compared to the other isozymes. Voles, characterized as strongly circadian rhythmic showed circadian variation in numbers of PKCalpha immunoreactive SCN neurons, while voles with weak or no circadian rhythmicity did not reveal such a circadian profile. PKCalpha immunoreactivity in acute SCN slices that were treated with a physiological dose of AVP was significantly lowered when compared with control slices. The intracellular messenger PKCalpha may reflect variation in locomotor behavior via the AVP system in the vole SCN. This system could play a key role in the vole SCN by mediating output of its circadian clock. PMID- 12637038 TI - The effects of corticotropin-releasing factor on the cortical EEG are reduced following adolescent nicotine exposure. AB - Although smoking is highly prevalent among adolescents, relatively little is known about the lasting neurobehavioral consequences associated with adolescent nicotine exposure. Prior studies from our laboratory suggest that adolescent nicotine exposure induces an anxiogenic profile in adult rats. Corticotropin releasing factor (CRF) systems are important modulators of anxiety and response to stress. Since acute nicotine administration has been shown to stimulate central CRF activity, the purpose of this study was to examine the effects of adolescent nicotine exposure on CRFs modulation of the cortical and hippocampal EEG in adult rats. Male Sprague-Dawley rats were exposed to nicotine (5 mg/kg/day) between postnatal days 35-40 using transdermal nicotine patches. Six weeks after nicotine exposure ended, the effects of intracerebroventricular administration of CRF (0.01-1.0 microg/5 microl) on EEG activity in the cortex and hippocampus were assessed in nicotine-exposed rats and age-matched control rats. The overall effects CRF were consistent with previous reports. CRF decreased low to moderate frequency EEG activity (1-32 Hz) and increased high frequency EEG activity (32-50 Hz). However, in nicotine-exposed rats the effects of CRF on the frontal and parietal cortical EEG were blunted by 30-50% compared to control rats. A similar pattern of decreased response to CRF was not observed in the hippocampus. These blunted effects of CRF on the cortical EEG suggest that long-term changes in systems responsive to CRF result from adolescent nicotine exposure. Given the role of CRF systems in behavioral responses to stress and anxiety, these data suggest that adolescent nicotine exposure may produce long term decreases in neurophysiological responses to stress. PMID- 12637039 TI - The role of the plasminogen activation cascade in glioma cell invasion: a review. AB - Tumour cell invasion is a dynamic process that depends on a co-ordinated series of biochemical events. This review discusses the role of the proteolytic enzyme system, the plasminogen activation cascade, in glioma cell invasion. PMID- 12637040 TI - A pathology of the animal spirits -- the clinical neurology of Thomas Willis (1621-1675). Part II -- disorders of intrinsically abnormal animal spirits. AB - Thomas Willis (1621-1675), author of the classical work Cerebri Anatome (1664), was arguably the father of the modern era of neurology. His clinical neurology, as described in his Pathologiae Cerebri (1667) and De Anima Brutorum (1672), was largely derived from personal observations and not from traditional authorities and was based around his concept of the animal spirits, a fictitious entity in many ways analogous to the present day idea of the nerve impulse. This concept allowed him to develop a pathology of the animal spirits which embraced the whole content of the clinical neurology and psychiatry of his times. The anatomical and physiological background to Willis' concepts of animal spirit dysfunction, and those disorders he regarded as due to disturbed function of intrinsically normal animal spirits, have been dealt with in the previous part of this paper. The disorders he attributed to intrinsically abnormal animal spirits, dealt with in this part of the paper, comprised two categories. In one, the animal spirits possessed explosive properties, whilst in the other the abnormalities were non explosive in their nature. The former category included epilepsy, hysteria and hypochondriasis, whilst the latter included mainly disorders now considered psychiatric e.g. delirium, melancholy, madness and stupidity. Willis' ideas about the pathogenesis of nervous system disorder seem never to have been generally accepted, partly because they appeared at a time when others were increasingly calling into question the existence of the animal spirits. Nevertheless, Willis' attempt to record and interpret all nervous system disease on the basis of disorder of function of a single underlying mechanism represents a formidable synthetic intellectual endeavour on the part of a very busy physician. PMID- 12637041 TI - Current intracerebral haemorrhage management. AB - Primary intracerebral haemorrhage (ICH) refers to spontaneous bleeding from intraparenchymal vessels. It accounts for 10-20% of all strokes, with higher incidence rates amongst African and Asian populations. The major risk factors are hypertension and age. In addition to focal neurological findings, patients may present with symptoms of elevated intracranial pressure. The diagnosis of ICH can only be made through neuro-imaging. A CT scan is presently standard, although MRI is increasingly important in the evaluation of acute cerebrovascular disease. A significant proportion of intracerebral haematomas expand in the first hours post ictus and this is often associated with clinical worsening. There is evidence that the peri-haematomal region is compromised in ICH. This tissue is oedematous, although the precise pathogenesis is controversial. An association between elevated arterial pressure and haematoma expansion has been reported. Although current guidelines recommend conservative management of arterial pressure in ICH, an acute blood pressure lowering trial is overdue. ICH is associated with a high early mortality rate, although a significant number of survivors make a functional recovery. Current medical management is primarily aimed at prevention of complications including pneumonia and peripheral venous thromboembolism. Elevated intracranial pressure may be treated medically or surgically. Although the latter definitively lowers elevated intracranial pressure, the optimal patient selection criteria are not clear. Aggressive treatment of hypertension is essential in the primary and secondary prevention of ICH. PMID- 12637042 TI - Visual dysfunction in artists. AB - The representation of the world provided by an artist is often modified by abnormalities in the visual pathways. Some artists have used these distortions to create a particular style. Controversy surrounds earlier artists whose visual function was not well documented historically. Drugs and psychiatric disease may also modify the artist's imagery. PMID- 12637043 TI - Endoscopic assisted microneurosurgery for cerebral aneurysms. AB - While there have been many advances in the field of microneurosurgery, the clipping of aneurysms remains an intricate procedure. Technical complications include residual aneurysm, perforator injuries, parent artery occlusion and cranial nerve injuries. The neuroendoscope is a useful tool and adjunct to the microsurgical clipping of these aneurysms. We study the usefulness of the neuroendoscope in enhancing visualisation during surgery. Twenty-four cases of ruptured cerebral aneurysms were operated on over a duration of 6 months in which a 1mm diameter rigid endoscope was used. We discuss our preliminary results and examine the advantages of the neuroendoscope. These include the ability to look around corners and behind obstructions. While this provides an additional view to the surgeon, the high magnification gives good definition of the surrounding structures. With less brain retraction, smaller operative exposures and yet better visualisation offered, neuroendoscopy may reduce operative morbidity. PMID- 12637044 TI - Progression in acute ischemic stroke: frequency, risk factors and prognosis. AB - BACKGROUND AND PURPOSE: The aim of this study was to investigate the frequency, possible predictive factors and the prognosis of deteriorating ischemic stroke. METHODS: A total of 266 stroke patients who presented within 24h of onset were enrolled. Clinical deterioration was defined as a decrease of > or =1 points in the Canadian Neurological Scale (CNS). Rankin Score (RS) was performed at discharge and at six months. RESULTS: Of the 266 patients studied, 26 (9.8%) worsened. Involvement of posterior circulation (odds ratio (OR) 3.16) and noncardioembolic infarction (OR 4.5) were found to be independently associated with neurological worsening. Death occurred in 19.2% of progressive (P) and in 4.16% of nonprogressive (NP) groups. Functional outcome was worse in the P than in NP patients at discharge and at sixth months. CONCLUSIONS: Involvement of posterior circulation and noncardioembolic subtypes of infarct independently affect neurological progression in acute ischemic stroke. Clinical deterioration significantly worsens the prognosis. PMID- 12637045 TI - Unilateral pallidotomy for Parkinson's disease disrupts ocular fixation. AB - Although some motor functions of the basal ganglia have been well studied, the oculomotor functions are not well established. We studied eye movements in patients with Parkinson's disease (PD) undergoing pallidotomy to assess the role of the globus pallidus interna (GPi) in oculomotor control. Horizontal visually guided, gap and predictive saccades as well as ocular fixation were studied in patients with advanced PD before and 1 month after unilateral pallidotomy, and in healthy controls on two occasions 1 month apart. There was no difference in saccadic latency or accuracy, the number of saccadic anticipations or the ability to generate predictive saccades between the two assessments for either patients or controls. The number and amplitude of square wave jerks during ocular fixation however increased significantly in patients after pallidotomy. The results imply altered function of frontal or prefrontal cortical regions involved in ocular fixation resulting from a disruption to inhibitory pallidal influences on thalamocortical projections. The posteroventral GPi however appears not to be involved in externally controlled or predictive saccadic function. PMID- 12637046 TI - Prevalence of cerebral vascular amyloid-beta deposition and stroke in an aging Australian population: a postmortem study. AB - Cerebral amyloid angiopathy (CAA) is a putative risk factor for lobar cerebral haemorrhage and infarction in the elderly. However, the prevalence of stroke in a population with CAA is not known. Amyloid-beta immunohistochemistry was used to assess CAA prevalence as a function of age, and the relationship between CAA and stroke in 100 individuals aged 50-91 years who died unexpectedly and had a Coroner's postmortem. Blocks were taken from several cortical areas and from areas of infarction or haemorrhage. Parenchymal Abeta was first found in the 6th decade, whereas vascular Abeta did not appear until the 7th decade. The prevalence of both vascular and parenchymal Abeta increased with age to a maximum in the 9th decade. The age at onset of vascular Abeta deposition was similar to that in an English study of CAA but a decade later than in Japanese studies. There was no association between the presence of vascular Abeta and cerebral haemorrhage or infarction. The findings indicate differences in the time-course of vascular and parenchymal Abeta deposition with age, as well as racial differences. The lack of association between vascular Abeta and cerebral haemorrhage or infarction indicates that, in the present population, CAA was usually asymptomatic. PMID- 12637047 TI - Results of surgical treatment for cerebral metastases. AB - Cerebral metastases are commonly encountered, with an incidence ranging from 20 to 30% of cancer cases. Medical and radiation therapy confer median survival ranging from 3 to 6 months only. A retrospective review of patients undergoing surgery with radiotherapy from January 1997 to January 2000 involved 26 patients (13 males, 13 females), with a mean age of 53.1 years (range 30-69 years). The mean follow-up was 15.8 months (range 1.5-27 months). The primary cancer was breast in nine cases, eight were lung cancers, six colonic cancers, two unknown primaries and one was a soft tissue sarcoma. Patients with extracranial secondaries numbered 17 (61.5%). Patients stayed on average for 15.4 days. No intracranial complications occurred postoperatively. No mortalities occurred in the first month and 11 patients (42.3%) were alive at 1 year. Two patients underwent a radiosurgical boost and six patients underwent adjuvant chemotherapy. Overall median survival was 9 months (95% confidence interval 7.3-12.9 months). Factors which may favour longer survival were female sex (median of 12 vs 6.1 months), younger patients (median of 12 vs 5.8 months) and breast cancer (median of 9 months). Surgical treatment appears beneficial, with subgroups such as breast cancer possibly doing better. PMID- 12637048 TI - Cerebral blood flow and dynamic cerebral autoregulation during ethanol intoxication and hypercapnia. AB - More than one-third of patients diagnosed with head injury are intoxicated with ethanol. Most clinical and animal studies have shown alcohol to have a deleterious impact in the setting of cerebrovascular trauma; however, there are also data showing neuroprotective effects in low ethanol doses. Human studies using imaging modalities suggest that small doses of alcohol produce cerebral vasodilatation and higher doses cerebral vasoconstriction. The aim of this study was to investigate the effect of ethanol intake on dynamic cerebral autoregulation and velocities in the middle cerebral arteries, and compare these changes with the effects of hypercapnia. Dynamic cerebral autoregulation and cerebral blood flow velocities were analysed before and after alcohol intake (1.1 g/kg of body weight) in six adult volunteers. Cerebral blood flow velocities in both middle cerebral arteries were monitored continuously by transcranial Doppler. A value for dynamic cerebral autoregulation was calculated from the rate of increase in middle cerebral artery velocities after a rapid-step decrease in arterial blood pressure. A sudden decrease in blood pressure was achieved by the release of previously inflated large blood pressure cuffs around the subject's thighs. Three volunteers were also tested before alcohol intake with CO(2) challenge (breathing 6% CO(2)) during the autoregulation procedure. Blood alcohol level reached 90 mg/dl approximately 60 min after ethanol ingestion. Cerebral blood velocities increased by 8% from baseline for uncorrected end-tidal (et) CO(2) and by 24% for correction to et CO(2)=40. Dynamic cerebral autoregulation measured as an autoregulation index decreased from 4.3+/-1.3 to 2.9+/-1.1 (p=0.089), which did not reach statistical significance. During hypercapnic conditions, dynamic cerebral autoregulation dropped from 4+/-0.8 to 0.9+/-0.9. In conclusion, mild alcohol intoxication caused cerebral vasodilatation with a subsequent increase in cerebral blood flow of 8-24%. Dynamic cerebral autoregulation was not found to be significantly impaired by ethanol. Hypercapnia almost completely destroys the physiological autoregulatory mechanism. A mild hyper-ventilation to etCO(2)=34-36 may be a compensatory contra-measure for ethanol-induced vasodilatation in the setting of head trauma. PMID- 12637049 TI - Multilevel subtotal corpectomy and interbody fusion using a fibular bone graft for cervical myelopathy due to ossification of the posterior longitudinal ligament. AB - A retrospective study of the outcomes of multilevel anterior decompression and interbody fusion for cervical myelopathy due to ossification of the posterior longitudinal ligament (OPLL) was performed to both investigate the long-term results and assess the cause of late deterioration. Twenty-seven patients (mean age, 58.1 years) underwent this procedure and were followed for at least 5 years. The severity of the clinical symptoms was described using the scoring system for cervical myelopathy proposed by the Japanese Orthopaedic Association (JOA score). The average preoperative JOA score was 7.7, and the score at final follow-up was 13.4 with a recovery rate of 62.0%. A delayed deterioration was attributed to a thoracolumbar lesion other than a compromising alteration of the cervical spine. Consequently, this method of treatment for OPLL could stop the progress of ossification and keep a physiological cervical alignment and thus provide good long-term results. PMID- 12637050 TI - The use of mild hypothermia for patients with severe vasospasm: a preliminary report. AB - The purpose of this study was to determine the effect of mild hypothermia on cerebral ischaemia due to severe vasospasm, which was refractory to medical and intravascular treatments and to assess the brain protection of this treatment in patients who underwent delayed aneurysm clipping after presenting with ischaemic neurological deficits. Mild hypothermia (32-34 degrees C of brain temperature) was employed in two groups: (1) Patients (Hunt and Kosnik grades I to II) who showed progressive neurological deficits due to vasospasm and did not respond to conventional therapy (Group 1) and (2) Patients who received delayed aneurysm clipping after presenting with ischaemic neurological deficits due to vasospasm (Group 2). Seven of 8 patients in both Groups showed a favorable outcome with mild hypothermia (good recovery in 5 and moderate disability in two patients). Mild hypothermia is considered to be effective on critical cerebral ischaemia due to vasospasm even after failure to response the conventional therapies and to provide brain protection in delayed aneurysm clipping. PMID- 12637051 TI - Agreement between experts: an answer, but is it the answer for you? AB - The question of how should agreement between experts be measured, when their opinions are expressed on an ordinal scale, is discussed. The example used is that of quality of articles submitted to Journal of Clinical Neuroscience. Polychoric correlation is a useful concept, and there are now computer programs available to estimate this. However, it is also emphasised that the definition of agreement is a matter of choice, and the choice should be made carefully: even error-free measurements should not automatically be assumed to possess all the properties of numbers, and ordinal categories bring further difficulties. PMID- 12637053 TI - Anterior pituitary hyperplasia due to primary autoimmune hypothyroidism. PMID- 12637054 TI - The effects of calcium channel antagonist nimodipine on end-plate vascularity of the degenerated intervertebral disc in rats. AB - The vascular channels at the end-plate of the intervertebral disc are very important in maintaining a healthy disc. With age, a reduction of the nutrition of the avascular nucleus pulposus is inevitable. On the other hand the calcium channel antagonist nimodipine has been shown to have a positive effect on blood flow in the region of the vertebral end-plate. To evaluate the effects of nimodipine on the end-plate vascularity in the degenerative discs, we have produced an experimental disc degeneration and evaluated the radiological and histopathological features of the end-plate of the degenerated discs. Adult rats were divided into 3 groups: control (n=5), operated degeneration (n=5), and nimodipine treatment (n=5). Using a posterior approach, a cut was made parallel to the end-plates in the posterior annulus fibrosus in 5 consecutive intervertebral discs between the 5th and 10th vertebral segments of the tails of adult Swiss Albino rats. At 8 weeks, 5 of these animals were treated with nimodipine. In each experimental group, 1 animal was examined using computed tomography (CT) to study the density of the cartilage end-plate of the disc. Then, the animals were sacrificed for subsequent histopathological evaluation. We found that the vascular channel counts and percentage areas from animals treated with nimodipine were higher than from both the non-operative control and operated degeneration groups, although these were not statistically different. Accordingly, the profile of the density histogram in the nimodipine-treated group showed a wide plateau, indicating an increase in the vascularity in this region. From our results, we suggest that nimodipine enhances vascularisation of the cartilage end-plate in the disc. It is possible that the increased proportion of vascular contacts at the end-plate has a beneficial effect in the nutrition of the disc. However, further experimental studies will be needed to determine the validity of this statement in animals or human beings. PMID- 12637056 TI - The accuracy and safety of image-guidance system using intraoperative fluoroscopic images: an in vitro feasibility study. AB - The "virtual fluoroscopy" is a new navigation system that combines image-guided surgery and C-arm fluoroscopy. The purpose of this study was to evaluate the accuracy of this system in the case of pedicle screwing. The angular deviation in the axial and sagittal planes was measured using the bodies of 50 vertebral models. Another type of model was used to evaluate the positional deviation (two dimension and three dimension). The mean angular deviation on the axial plane was 1.3 degrees and that on the sagittal plane was 1.0 degree. The positional deviation was not recognised. This system provides quite accurate information. Using this system, the surgeon can insert surgical instruments, such as pedicle screws, while confirming the real time virtual image on the monitor displaying the surgical field, including the position and direction of the instruments, without being disturbed by his own hands or instruments and without radiation exposure. PMID- 12637055 TI - Differences in responses to nociceptive stimulation of the oral and aboral oesophagus. AB - The present study examined the role of vagal innervation of the rat esophagus in nociception. Electromyographic recordings from neck muscles were used as indicators of pseudoaffective reactions in lightly anaesthetized animals; responses were obtained during mechanical (distension) and chemical (HCl) nociceptive stimulation of the upper cervical (4.5 cm from incisors) and midthoracic (7 cm from incisors) esophagus. Compared to midthoracic esophageal stimulation, stimulation of the upper esophagus produced more vigorous responses. Bilateral transections of the cervical vagus and superior laryngeal nerves led to a gradual diminution of responses to upper stimulation, while the same manoeuvre increased responses to lower esophagus stimulation. These results suggest a contribution of vagal afferents to pseudoaffective responses elicited by noxious stimulation of the upper esophagus. PMID- 12637057 TI - Microendoscopic discectomy (MED) for lumbar disc prolapse. AB - Microendoscopic discectomy (MED), which combines traditional lumbar microsurgical techniques with endoscopy, is being used as a minimally invasive procedure for lumbar disc herniation. We reviewed 30 patients who underwent MED at our institution and compared their outcome with that of patients subjected to the conventional method. Laboratory data suggested that MED was less invasive surgery. Moreover, MED allowed an early return to work. However, the difficulties of this endoscopic procedure were evident, because of the limited exposure and two-dimensional video display. The potential injury of the nerve root and prolonged surgical time remain as matters of serious concern. To overcome this problem, we used an operative magnifying glass during surgery and this helped us to accomplish the procedure comfortably. We recommend the use of an operative magnifying glass in the early stage of the introduction of MED, for it is quite useful to identify the three-dimensional relationships of the structures. PMID- 12637058 TI - Focal magnetoencephalographic spikes in the superior temporal plane undetected by scalp EEG. AB - Temporal lobe spikes were detected by magnetoencephalography (MEG), but not by standard scalp electroencephalography (EEG), in a patient with intractable complex partial seizures. Simultaneous recording of scalp EEG and MEG revealed 2 different types of spike discharges: sporadic single spikes detected by both EEG and MEG which were localised diffusely in the right temporal lobe; and rhythmic MEG spike discharges that were not detected by scalp EEG, focally localised in the posterior part of the superior temporal plane. The tangential current orientation to the scalp may explain the different sensitivity of scalp EEG and MEG to rhythmic discharges. This study shows the unique sensitivity of MEG to epileptic activity in the superior temporal plane. PMID- 12637059 TI - Neuropsychological manifestations in a case of bilateral thalamic infarction. AB - Neuropsychological manifestation has been reported with lesions of the anterior and non-specific thalamic nuclei and mammilothalamic tract (MMT). These have been reported in the setting of arterial infarction and/or haemorrhage. Cerebral venous sinus thrombosis (CVT) is a rare cause of brain infarction. It occurs in the setting of oral contraceptive administration or pregnancy. Inherited thrombophilias are documented risk factors. The most frequent being heterozygous factor V Leiden mutation. We report a single case of bilateral thalamic infarction due to cerebral vein and sinus thrombosis. Clinically the case manifested with memory impairment and dysexecutive symptoms. Predisposing factor for venous thrombosis was a homozygous factor V Leiden mutation. The patient was treated with anticoagulation and made a good recovery. PMID- 12637060 TI - Dural arteriovenous fistulae causing pseudotumour cerebri syndrome in an elderly man. AB - The clinical presentation of dural arteriovenous fistulae (DAVF) is dependent on their location and the nature of their venous drainage. The latter plays a critical part in determining whether or not the fistula gives rise to intracranial hypertension, which is present in only a minority of cases. We present a case of the pseudotumour cerebri syndrome in an elderly man with bilateral intracranial DAVF supplied by the occipital arteries. Cerebral angiography was required for definitive diagnosis, and to characterise the abnormal venous drainage. The pathophysiology of intracranial hypertension in DAVF is discussed. PMID- 12637061 TI - Softly spoken strokes: two patients with marked hypophonia as a feature of strokes involving the anterior thalamus. AB - Two patients with strokes involving thalamic structures are described. Both displayed marked hypophonia as part of their clinical syndromes. This speech disorder and its possible localizing value are discussed. PMID- 12637062 TI - Compression of the ulnar nerve at the elbow by an intraneural ganglion. AB - A rare cause of ulnar nerve compression at the elbow is presented in this report. A 42 year old right-handed mechanic developed subacute, progressive numbness, tingling and weakness in his right hand. Electrophysiologic studies demonstrated a severe conduction block affecting the ulnar nerve in the retrotrochlear groove but without any sign of major axonal loss. His hand functions were carefully studied prior to surgery. While fine motor tasks were not affected, the hand strength was markedly diminished. At surgery, a 1-cm diameter intraneural ganglion at the site of the conduction block was found and excised. The patient made a dramatic recovery within 6 weeks post-surgery. The conduction block completely resolved and the hand functions also returned to normal. This and other reported cases point to the importance of early diagnosis and intervention. PMID- 12637063 TI - Intracranial migration of Foley catheter--an unusual complication. AB - Profuse nasal bleeding in cases of severe head injury is mostly associated with facial and skull base fractures and various methods have been tried to control such bleeding. Anterior nasal packing, producing a tamponade effect with a Foley catheter and other inflatable balloon devices are time proven methods. We describe a case of severe head injury with such fractures where a Foley catheter tamponade was attempted to control the severe nasal bleeding and the CT brain scan revealed inadvertent malposition of the catheter into the cranial cavity. This rare entity is described to highlight the potential danger associated with a proven method of controlling severe nasal bleeding and suggestions are outlined as to how to avoid such a mishap, along with a brief review of the literature. PMID- 12637064 TI - Recurrent subarachnoid haemorrhage from a de novo basilar bifurcation aneurysm: a case report. AB - Well documented cases of posterior circulation de novo aneurysms are rare. We report a patient who recently presented following a third episode of aneurysmal subarachnoid haemorrhage. Subsequent cerebral angiography revealed a basilar bifurcation aneurysm. This was absent on previous angiographic studies. The current literature on de novo aneurysms is reviewed. The implications for screening are considered. PMID- 12637065 TI - Simultaneous occurrence of meningioma and glioma in brain: report of two cases. AB - Two patients who presented with a simultaneous occurrence of meningioma and glioma are described. The authors describe their neuroradiological findings, best surgical treatment of this association, aspects of pathology and aetiology, along with a review of literature. PMID- 12637066 TI - Optochiasmal arachnoiditis following cotton wrapping of anterior communicating artery aneurysm treated by surgical removal of granuloma. AB - We report the case of a 50 year old female who presented with visual disturbance due to optochiasmal arachnoiditis and foreign body granuloma 9 months after cotton wrapping for ruptured anterior communicating artery (AcomA) aneurysm. Magnetic resonance imaging (MRI) revealed enhanced mass lesion around AcomA complex and hyperintense signal on optic chiasm and right optic tract by fluid attenuated inversion recovery image. Despite the repeated steroid pulse therapy, she deteriorated and MRI showed expansion of the granulomatous lesion over 5 months. Surgical removal of foreign body granuloma resulted in marked improvement of visual disturbance as well as of the MRI findings. We conclude that the use of cotton sheet close to the optic nerve should be avoided, and that surgical removal of the granuloma would be the optimal choice especially for the patient in whom steroid therapy fails to improve clinical symptoms. PMID- 12637067 TI - Fenestration of the internal carotid artery associated with arterial dissections. AB - An increasing number of cases with fenestration that is frequently associated with aneurysms have been reported, largely in the vertebral and basilar arteries and less commonly in the middle and anterior cerebral arteries by angiography as well as by autopsy. However, the anomaly of the internal carotid artery associated with bilateral arterial dissections is exceedingly rare. This represents the first reported case of fenestration of the internal carotid artery associated with arterial dissections. The embryological basis and clinical significance of this anomalous condition are discussed and treatment for arterial dissection is presented. PMID- 12637068 TI - Pituitary apoplexy and inappropriate ADH secretion. AB - Pituitary apoplexy may be associated with visual deterioration that may be severe in some cases. Misdiagnosis of this condition is not uncommon, resulting in delayed treatment, which may adversely affect the outcome and visual prognosis. We present a case of pituitary apoplexy, who presented with features of syndrome of inappropriate anti-diuretic hormone secretion (SIADH) and monocular blindness. He had remarkable improvement to normal vision along with normalisation of serum sodium following emergency trans-sphenoidal surgery. PMID- 12637070 TI - Clear cell meningioma: case report and literature review. AB - The clear cell meningioma is rare and a recently described histologic variant of meningioma. The most interesting aspect of clear cell meningioma is the high recurrence rate and agressiveness. Until now 17 intracranial clear cell meningioma cases had been reported in the English language literature. We present 2 new cases of clear cell meningioma which is discussed with the relevant literature. PMID- 12637069 TI - En-plaque ventricular ependymoma. AB - An extremely unusual case demonstrating rare behaviour of an ependymoma is presented. The tumour spread in an en-plaque fashion and involved the ependymal lining of the entire ventricular system. The patient, a 56-year-old male, presented with symptoms of raised intracranial pressure and truncal ataxia. The management issues are analyzed on the basis of a brief literature survey. PMID- 12637071 TI - Solitary plasmacytoma with intracranial intraorbital and, paranasal sinus extension. AB - Intracranial solitary plasmacytomas (ICSPs) are extremely rare tumours in neurosurgical practice, and are often misdiagnosed preoperatively. Here we present a solitary intracranial plasmacytoma with orbital, nasal and paranasal sinus extension. A subtotal excision of the tumour was performed and the complete response was seen after postoperative radio-chemotherapy. The neuroradiological and neurosurgical features of the case are discussed with the pertinent literature. PMID- 12637072 TI - A neurofibromatosis type 1 patient with severe kyphoscoliosis and intrathoracic meningocele. AB - The patient presented with neurofibromatosis and a dystrophic kyphoscoliosis around the cervico-thoracic junction. When the patient was 59 years old, he started to suffer from dyspnea caused by an intrathoracic meningocele in the upper left thoracic cavity. A wide laminectomy from T2 to T5 was performed and the meningocele was resected. Although the dyspnoea disappeared postoperatively, the patient started to neurologically deteriorate. Laminectomy alone caused instability around the apex of the kyphosoliosis and spinal cord compression. Halo cast was applied and brought remarkable recovery of neurologic deficits. This result encouraged us to perform posterior fusion in situ from C3 to L2 with bone graft from the iliac crests and the Luque technique in conjunction with the Isola system. This resulted in the patient being able to walk again. The removal of the posterior element predisposes the patient to unstable postlaminectomy kyphosis and removes valuable bone stock required for posterior spinal fusion. For this reason, spinal fusion should have been conducted during surgery for the patient's meningocele. PMID- 12637073 TI - Obliteration of giant corpus callosum AVM with linac based stereotactic radiosurgery. AB - Successful management of large AVMs is difficult. Traditionally they are considered less responsive and even unsuitable for radiosurgery. This case report demonstrates total angiographic obliteration in a complex, large corpus callosum AVM (volume 36.52 cc) in a 39-year-old male. Stereotactic radiation was delivered with a Linear Accelerator using ultra conformal treatment planning. Large volume AVMs can be subjected to stereotactic radiosurgery if the shape and location makes it possible to deliver an adequate radiation dose. PMID- 12637074 TI - A rare case of dysembryoplastic neuroepithelial tumour in occipital lobe presenting with only headache. AB - We report an unusual case of dysembryoplastic neuroepithelial tumour (DNT) located in the occipital lobe presenting with only headache. A 31 year old woman presented with headache. She had no history of epilepsy and neurological examination revealed no abnormal findings. Computed tomography (CT) scanning revealed a multilobulated mass lesion with calcification in the right occipital lobe. Magnetic resonance (MR) imaging demonstrated a heterogeneously enhanced mass with hypointense signals on T1- and hyperintense signals on T2-weighted images. The lesion was totally resected and histopathologically diagnosed as DNT. Physicians must bear in mind that DNT may occur in the occipital lobe and present with only mass effect. PMID- 12637075 TI - Nipah virus outbreak in Malaysia. AB - Nipah virus, a novel paramyxovirus, closely related to Hendra virus emerged in northern part of Peninsular Malaysia in 1998. The virus caused an outbreak of severe febrile encephalitis in humans with a high mortality rate, whereas, in pigs, encephalitis and respiratory diseases but with a relatively low mortality rate. The outbreak subsequently spread to various regions of the country and Singapore in the south due to the movement of infected pigs. Nipah virus caused systemic infections in humans, pigs and other mammals. Histopathological and radiological findings were characteristic of the disease. Fruitbats of Pteropid species were identified as the natural reservoir hosts. Evidence suggested that climatic and anthropogenic driven ecological changes coupled with the location of piggeries in orchard and the design of pigsties allowed the spill-over of this novel paramyxovirus from its reservoir host into the domestic pigs and ultimately to humans and other animals. PMID- 12637078 TI - Hepatitis C virus genotypes in Hungarian and Austrian patients with chronic hepatitis C. AB - Hepatitis C virus (HCV) genotypes are relevant to epidemiological questions, vaccine development, and clinical management of chronic HCV infection. The aim of this study was to determine HCV genotypes of South Hungarian and Southeast Austrian patients with chronic hepatitis C. Results were obtained by the largely automated TruGene HCV 5'NC Genotyping Kit (Visible Genetics, Toronto, Ontario) and by phylogenetic analysis. All of the 20 Hungarian patients and 15 out of 20 Austrian patients were infected with genotype 1. The remaining Austrian patients were infected with genotypes 3 or 2. With the commercially available assay, it was not possible to determine the HCV subtype in a total of three patients. The TruGene HCV 5'NC Genotyping assay for the determination of HCV genotypes proved to be useful for a high-throughput routine diagnostic laboratory. PMID- 12637076 TI - Herpes zoster guideline of the German Dermatology Society (DDG). AB - Varicella zoster virus (VZV) causes varicella (chickenpox), remains dormant in dorsal root and cranial nerve ganglia and can be reactivated as a consequence of declining VZV-specific cellular immunity leading to herpes zoster (shingles). Patients older than 50 years of age affected by herpes zoster may suffer a significant decrease of quality of life. These patients and immunocompromised individuals are at increased risks for severe complications, involving the eye, the peripheral and the central nervous system (prolonged pain, postherpetic neuralgia). Such complications occur with and without cutaneous symptoms. The German Dermatology Society (DDG) has released guidelines in order to guarantee updated management to anyone affected by herpes zoster. Diagnosis is primarily clinical. The gold standard of laboratory diagnosis comprises PCR and direct identification of VZV in cell cultures. Detection of IgM- and IgA-anti VZV antibodies may be helpful in immunocompromised patients. Therapy has become very effective in the last years. Systemic antiviral therapy is able to shorten the healing process of acute herpes zoster, to prevent or to alleviate pain and other acute and chronic complications, particularly, when given within 48 h to a maximum of 72 h after onset of the rash. Systemic antiviral therapy is urgently indicated in patients beyond the age of 50 years and in patients at any age with herpes zoster in the head and neck area, especially in patients with zoster ophthalmicus. Further urgent indications are severe herpes zoster on the trunk and on the extremities, herpes zoster in immunosuppressed patients and in patients with severe atopic dermatitis and severe ekzema. Only relative indications for antiviral therapy exist in patients younger than 50 years with zoster on the trunk and on the extremities. In Germany acyclovir, valacyclovir, famciclovir and brivudin are approved for the systemic antiviral treatment of herpes zoster. These compounds are all well tolerated by the patients and do not differ with regard to efficacy and safety. Brivudin has a markedly higher anti VZV potency than oral acyclovir, valacyclovir and famciclovir and thus offers a simpler dosing regimen. It must be given only once daily during 7 days in comparison to three and five times dosing per day of valacyclovir, famciclovir and acyclovir, respectively. Brivudin is an antiviral agent with no nephrotoxic properties, which is an advantage when compared to acyclovir. The most important aim of therapy of herpes zoster is to achieve painlessness. Appropriately dosed analgesics in combination with a neuroactive agent (i.e. amitriptylin) are very helpful when given together with antiviral therapy. The additive therapy with corticosteroids may shorten the degree and duration of acute zoster pain, but has no essential effect on the development of postherpetic neuralgia, which is a very difficult condition to treat. Thus early presentation to a pain therapist is recommended in specific cases. PMID- 12637079 TI - Point mutations induced by foscarnet (PFA) in the human cytomegalovirus DNA polymerase. AB - BACKGROUND: In vitro selection of viruses with decreased drug susceptibility is a useful tool for mapping drug resistance-associated alterations, evaluating cross resistance profiles, and elucidating molecular mechanisms of antiviral activity. OBJECTIVES: To provide data on mechanisms of selective drug action and features of drug resistance that may be clinically important. STUDY DESIGN: Foscarnet (PFA) and ganciclovir (GCV) were used to induce mutants of the human cytomegalovirus (HCMV) Towne strain. RESULTS: Three new mutations, selected in the presence of PFA, were identified with single base substitutions resulting in T419M, Q578H, and L773V in conserved regions of the HCMV DNA polymerase. None of these mutations have been reported previously. These mutations conferred resistance to PFA but did not change the susceptibility to GCV. A mutant was selected in the presence of GCV. This GCV-selected mutant had no mutation in the UL54 but had an amino acid alteration at codon M460V of UL97, which conferred resistance to GCV. All the mutants had the same growth phenotype as the parental laboratory strain Towne. CONCLUSIONS: We have determined three novel alterations in HCMV DNA polymerase inducing reduced susceptibility to PFA. None of these alterations changed the growth phenotype of the parental virus. PMID- 12637080 TI - Comparison of the effects of acute influenza infection and Influenza vaccination on HIV viral load and CD4 cell counts. AB - PURPOSE: Influenza vaccination is recommended for HIV-infected patients, although the efficacy is not clear. Prior studies have yielded differing results with regard to the effects of influenza vaccination on HIV viral load and CD4 cell counts. The effects of acute influenza on HIV viral replication and CD4 cell counts have not been well described. We sought to assess the effect of influenza infection and vaccination on HIV viral load and CD4 cell counts. SUBJECTS AND METHODS: All cases of influenza occurring in HIV-infected individuals over 3 years at a large county hospital were reviewed. For the year 1997-1998, data on all HIV clinic patients who were vaccinated for influenza were recorded prospectively. In order to assess the effects of influenza infection (Group I) and vaccination (Group II) on HIV viral load and CD4 cell counts, values from before and after influenza infection or vaccination were compared to each other and to a matched control group not vaccinated and without influenza infection (Group III). RESULTS: Forty-three cases of influenza were diagnosed. Pre- and post-influenza viral load in Group I was not significantly different: 3.34 versus 3.49 log copies/ml (P=0.36). Viral load was unchanged in 22 of 37 patients, increased in ten patients and decreased in five patients. Similarly, pre- and post-vaccination viral load in Group II was not significantly different: 3.52 versus 3.66 log copies/ml (P=0.12). Thirty-four of 47 patients who received influenza vaccine had no significant change in viral load-viral load increased in ten patients and decreased in three patients. No significant CD4 cell count changes were noted following influenza infection or vaccination. In contrast, Group III patients experienced a small decline in viral load from 4.23 to 3.39 log copies/ml, P<0.05, while there was a trend towards an increase in CD4 cell counts (P=0.06). CONCLUSIONS: Following influenza infection or vaccination, most patients did not have a significant increase in HIV viral load or decrease in CD4 cell count. PMID- 12637081 TI - Discrimination between dog-related and vampire bat-related rabies viruses in Brazil by strain-specific reverse transcriptase-polymerase chain reaction and restriction fragment length polymorphism analysis. AB - BACKGROUND: There is a geographical overlap between the two main rabies epidemiological cycles maintained by dogs and vampire bats in Latin America. The geographical and temporal coincidence of rabies outbreaks of respective origins is not unusual in rural areas of Latin America. These circumstances make it difficult to discriminate the intraspecies and interspecies transmission pathways of rabies. OBJECTIVE: This study was conducted to develop techniques to discriminate dog-related and vampire bat-related rabies virus isolates (DRRV and VRRV, respectively) in Brazil. STUDY DESIGN: The 1396 nucleotides of the nucleoprotein gene of a total of 27 DRRV and VRRV were sequenced. Strain-specific (SS) primers were developed based on these sequences. Forty-nine rabies virus strains isolated from animals and humans in several parts of Brazil were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) with SS primers. These rabies viruses were also amplified by RT-PCR with general rabies primers and the PCR products were cut by three restriction enzymes, Blp I, Bsu36 I and BspE I. RESULTS: All the DRRV and VRRV were distinguished by RT-PCR with SS primers. The PCR products obtained from DRRV were cut at one site by Blp I, but not by Bsu36 I. The PCR products obtained from VRRV were cut at one or two sites by Bsu36 I, but not by Blp I. Blp I and Bsu36 I clearly discriminated DRRV and VRRV in restriction fragment length polymorphysim (RFLP) assays. The results of SS RT-PCR and RFLP were consistent. CONCLUSION: SS RT-PCR and RFLP assays have been developed for determining the origins of rabies virus isolates in Brazil. These assays are simple and rapid, and will be useful for identifying the rabies virus reservoirs of field isolates in Brazil, especially when used together. PMID- 12637082 TI - Clinical assessment of a generic DNA amplification assay for the identification of respiratory adenovirus infections. AB - BACKGROUND: respiratory adenoviruses are common, often resulting in serious sporadic and epidemic infections and impaired immunity can dramatically increase their severity. They are now thought capable of establishing latency. Diagnosis by culture is slow while direct antigen detection by immunofluorescence lacks sensitivity. Molecular diagnosis can be both rapid and sensitive but the genetic heterogeneity of adenoviruses poses problems. OBJECTIVES: to design a generic adenovirus nested polymerase chain amplification assay designed to be capable of detecting all respiratory adenoviruses. This was achieved through optimised thermal cycling and the development of a generic degenerate primer set targeting the adenovirus hexon gene. STUDY DESIGN: this was a cross-sectional study on 172 respiratory specimens from hospital-based patients, and one from a general practice, in Northern Ireland. A comparison was made between the amplification assay, virus culture and immunofluorescence. RESULTS: the nested polymerase chain reaction (nPCR) assay had a generic capacity for adenovirus detection and an analytical sensitivity of 6.4x10(2) copies/ml. Using an expanded gold standard (defined as a true positive or a true negative where a specimen was positive or negative by at least two of the study assays, respectively), PCR had a clinical sensitivity and specificity of 46/46 (100%) and 15/126 (91.3%), respectively. Patients with acute respiratory adenovirus infections were more likely to be male (chi(2), p=0.005) and to present with a fever (chi(2), p=0.02) than patients diagnosed with another respiratory virus. Co-infection was identified in 12/172 patients. CONCLUSIONS: the nested amplification assay proved highly sensitive in both the analytical and clinical settings for the detection of respiratory adenovirus infections. PMID- 12637084 TI - Identification of parental origin of cognate dsRNA genome segment(s) of rotavirus reassortants by constant denaturant gel electrophoresis. AB - Rotaviruses are the single most important etiologic agents of severe diarrhea in infants and young children worldwide. They possess a triple capsid morphology and a genome of 11 segments of double-stranded (ds) RNA. During the course of the development of various live, attenuated reassortant rotavirus vaccines, we often experienced difficulty in identifying the parental origin of certain genome segment(s) of a reassortant vaccine candidate. Various assays have been utilized for determination of the parental origin of reassortant virus genes, including polyacrylamide gel electrophoresis (PAGE), DNA and/or RNA hybridization assays and gene sequence analysis. The traditional PAGE is simple and easy to perform, however, it is common to find that certain cognate dsRNA segment(s) cannot be differentiated by this assay due to a high degree of sequence homology among different rotavirus strains. Constant denaturant gel electrophoresis (CDGE) is one of several methods that have been used to screen DNA fragments for small sequence changes or point mutations. By using the CDGE, we were successful in partially denaturing rotavirus dsRNA thereby changing the physical properties of the genome segment(s) in the gel and thus differentiating the cognate genome segment(s) of rotavirus reassortants. The CDGE provides a simple and reliable assay system for identification of parental gene origins of a rotavirus reassortant. PMID- 12637083 TI - Use of induced sputum for the diagnosis of influenza and infections in asthma: a comparison of diagnostic techniques. AB - BACKGROUND: Influenza (Flu) and respiratory syncytial virus (RSV) are important viral pathogens that cause lower respiratory tract infections and severe exacerbations of asthma. Molecular biological techniques are permitting a rapid and accurate diagnosis of infections caused by respiratory pathogens, and have typically been applied to upper respiratory samples. Sputum induction provides an opportunity to directly sample secretions from the lower respiratory tract. OBJECTIVES/STUDY DESIGN: To determine the role of induced sputum reverse transcription polymerase chain reaction (RT-PCR) in the detection of respiratory pathogens and compare this with detection using serology and immunofluorescent antigen (IFA) testing, we recruited 49 adults from emergency room with exacerbations of asthma. After a medical assessment and spirometry, sputum was induced using ultrasonically nebulised normal saline. Sputum was assayed using IFA and RT-PCR for flu and RSV. Flu serology was performed acutely and at convalescence, 4-5 weeks later. RESULTS: Influenza A or B was detected in 24% of the samples by PCR, significantly more than the nine cases detected using serology and the one case using IFA (P<0.05). RSV was detected in 37% of samples using PCR and 20% by IFA (P<0.05). CONCLUSION: The combination of induced sputum and RT-PCR provides a useful means of detecting respiratory infection. The technique is safe in both adults and children, and RT-PCR is more sensitive than conventional serology and IFA. The improved sensitivity of induced sputum RT-PCR also permits a more rapid diagnosis and the opportunity of early administration of effective treatments. PMID- 12637085 TI - Determination of enterovirus serotype inferred from sequence analysis of PCR products. AB - BACKGROUND: Enterovirus infections are common in neonates. Virus isolation is the only diagnostic method to confirm enterovirus serotype infections, however, is not always successful. OBJECTIVES: A new approach for the diagnosis of enterovirus infections was performed, using the reference strain inferred from sequence analysis of PCR products. STUDY DESIGN: Virus isolation, enterovirus RT PCR and sequence analysis were performed from clinical samples or stored sera from two neonates with fever and rash. Neutralizing test (NT) antibodies against prototype reference virus were measured in paired sera. RESULTS: Virus isolation was negative in both patients but the enterovirus genome was amplified in the acute phase sera obtained from the two patients. From the results of sequence analysis of 109 nucleotides located in the 5'-noncoding of the conserved region of enteroviruses, a high homology to echovirus types 25 and 30 was found. More than a 4-fold increase in NT antibodies against reference viruses was demonstrated in the acute and convalescent phase sera. They were confirmed as echovirus type 25 and 30 infection, respectively. CONCLUSIONS: These virological examinations are practical and useful for clinical settings for a diagnosis of enterovirus infections because of an insufficient positive rate in virus isolation. PMID- 12637087 TI - Optimisation of PCR reactions using primer chessboarding. AB - In-house polymerase chain reaction (PCR) assays are now an integral part of the work of most diagnostic microbiological laboratories. Despite the availability of commercial reagent 'master-mixes' of some PCR reagents, the optimisation of primers still poses a significant problem. Here, we describe a simple method to assess the concentration of primer needed in single round, multiplex, nested and 'real-time' PCR procedures. PMID- 12637088 TI - Improved molecular identification of enteroviruses by RT-PCR and amplicon sequencing. PMID- 12637086 TI - B19 virus infection in renal transplant recipients. AB - BACKGROUND: B19 virus infection with persistent anaemia has been reported in organ transplant recipients. Detection of B19 virus DNA in serum is the best direct marker of active infection. OBJECTIVE: The present study evaluated the incidence and clinical role of active B19 virus infection in renal transplant recipients presenting with anaemia. STUDY DESIGN: Forty-eight such recipients were investigated by nested PCR on serum samples. The controls were 21 recipients without anaemia. Active HCMV infection was also investigated as a marker of high immunosuppression. RESULTS AND CONCLUSIONS: In 11/48 (23%) patients B19 virus DNA was demonstrated in serum versus only 1/21 (5%) of the controls. Ten of these 11 patients had already been seropositive at transplantation and active infection occurred in eight of them during the first 3 months after transplantation. The remaining patient experienced a primary infection 9 months after transplantation. Eight (73%) of these 11 patients displayed a concomitant HCMV infection and four (36%) showed increasing serum creatinine levels but none developed glomerulopathy; 3/11 (27%) recovered spontaneously from anaemia whereas 8/11 (73%) needed therapy. In conclusion, the relatively high occurrence (23%) of B19 virus infection in patients presenting with anaemia, suggests that it should be considered in the differential diagnosis of persistent anaemia in renal transplant recipients. Presence of the viral DNA should be assessed early from transplantation and the viral load should be monitored to follow persistent infection and better understand the relation between active infection and occurrence of anaemia, and to assess the efficacy of IVIG therapy and/or immunosuppression reduction in clearing the virus. PMID- 12637089 TI - Comparison of primers for NLV diagnosis. PMID- 12637090 TI - Improvement of pulmonary absorption of cyclopeptide FK224 in rats by co formulating with beta-cyclodextrin. AB - FK224 is a cyclopeptide drug with a low aqueous solubility. Following oral administration to rats, poor absorption was observed due to proteolysis in the gastrointestinal tract. The objective of this study was to investigate the effect of the pulmonary route on the systemic absorption of FK224 in comparison with other administration routes, and to determine the bioavailability (BA) of FK224 following pulmonary administration in rats using various dosage forms. From absorption studies on the Polyethylene Glycol 400 solution given by various routes (intranasal, subcutaneous, intratracheal and intravenous as reference), it was shown that pulmonary administration was a potentially attractive route for FK224. In the pulmonary absorption studies, after administration of the aqueous suspension, the BA was reduced to 2.7% compared with 16.8% for the solution. However, beta-cyclodextrin (beta-CyD) was found to be an effective additive as far as improving the solubility of FK224 was concerned. The BA of the aqueous suspension containing beta-CyD was increased to 19.2%. Pressurized metered dose inhalers were prepared by formulating beta-CyD with various molar ratios of 1:0, 1:1 and 1:7 (FK224/beta-CyD), and the resulting BAs were 4.3%, 29.0% and 91.2%, respectively. It was observed that both the C(max) and AUC of FK224 were increased as the amount of beta-CyD increased. The plasma profiles showed sustained absorption. In conclusion, we have seen that the lung is a suitable route for absorption of FK224, and beta-CyD is an extremely effective additive as far as improving the pulmonary absorption of FK224 is concerned. beta-CyD or derivatives with various degrees of aqueous solubility are potential drug carriers for controlling pulmonary absorption. PMID- 12637091 TI - Preparation of poly-lactic acid microspheres containing the angiogenesis inhibitor TNP-470 with medium-chain triglyceride and the in vitro evaluation of release profiles. AB - TNP-470 (AGM-1470, 6-0-(N-chloroacetylcarbamoyl)-fumagillol), a derivative of fumagillin, is a promising angiogenesis inhibitor. However, as TNP-470 is very unstable in in vitro and in vivo, it has been difficult to verify its pharmacological efficacy in the clinical medicine. The preparation of a drug delivery system (DDS) in a microsphere form was studied for the stable inclusion and controlled release of TNP-470. Medium-chain triglyceride (MCTG) as an effective stabilizer and poly-lactic acid (PLA) as a biodegradable carrier were used for this purpose. The release of TNP-470 from the MCTG containing DDS continued for approximately 2 weeks, while the release of TNP-470 from the one without MCTG stopped after only 5 days. It was proved that TNP-470 could be released much more stable for much longer period from the MCTG containing DDS compared to the one without DDS. PMID- 12637092 TI - Micronized powders of a poorly water soluble drug produced by a spray-freezing into liquid-emulsion process. AB - The purpose of this paper is to investigate the influence of the emulsion composition of the feed liquid on physicochemical characteristics of drug-loaded powders produced by spray-freezing into liquid (SFL) micronization, and to compare the SFL emulsion process to the SFL solution process. Danazol was formulated with polyvinyl alcohol (MW 22,000), poloxamer 407, and polyvinylpyrrolidone K-15 in a 2:1:1:1 weight ratio (40% active pharmaceutical ingredient (API) potency based on dry weight). Emulsions were formulated in ratios up to 20:1:1:1 (87% API potency based on dry weight). Ethyl acetate/water or dichloromethane/water mixtures were used to produce o/w emulsions for SFL micronization, and a tetrahydrofuran/water mixture was used to formulate the feed solutions. Micronized SFL powders were characterized by X-ray diffraction, surface area, scanning and transmission electron microscopy, contact angle and dissolution. Emulsions containing danazol in the internal oil phase and processed by SFL produced micronized powders containing amorphous drug. The surface area increased as drug and excipient concentrations were increased. Surface areas ranged from 8.9 m(2)/g (SFL powder from solution) to 83.1 m(2)/g (SFL powder from emulsion). Danazol contained in micronized SFL powders from emulsion and solution was 100% dissolved in the dissolution media within 2 min, which was significantly faster than the dissolution of non-SFL processed controls investigated (<50% in 2 min). Micronized SFL powders produced from emulsion had similar dissolution enhancement compared to those produced from solution, but higher quantities could be SFL processed from emulsions. Potencies of up to 87% yielded powders with rapid wetting and dissolution when utilizing feed emulsions instead of solutions. Large-scale SFL product batches were manufactured using lower solvent quantities and higher drug concentrations via emulsion formulations, thus demonstrating the usefulness of the SFL micronization technology in pharmaceutical development. PMID- 12637093 TI - In-situ-micronization of disodium cromoglycate for pulmonary delivery. AB - Drug particle properties are critical for the therapeutic efficiency of a drug delivered to the lung. Jet-milling, a commonly used technique for micronization of drugs, has several disadvantages such as a non-homogeneous particle size distribution, and unnatural, thermodynamically activated particle surfaces causing high agglomeration. For pulmonary use in a dry powder inhaler, in addition to a small particle size, good de-agglomeration behaviour is required. In this study disodium cromoglycate is prepared in situ in a respirable particle size by a controlled crystallization technique. First the drug is dissolved in water (4%) and precipitated by a solvent change method in the presence of a cellulose ether (hydroxypropylmethylcellulose) as a stabilizing hydrocolloid. By rapidly pouring isopropyl alcohol into the drug solution in a 1:8 (v/v) ratio, the previously molecularly dispersed drug is associated to small particles and stabilized against crystal growth in the presence of the hydrophilic polymer. This dispersion was spray-dried. The mean particle size of the drug was around 3.5 microm and consequently was in the respirable range. The in-situ-micronized drug powder was tested for its aerodynamic behaviour and compared with jet-milled drug powder and with commercial products using the Spinhaler, the Cyclohaler, and the FlowCaps-Inhaler as model devices. The fine particle fraction (FPF) (<5 microm) was increased from 7% for the jet-milled drug to approximately 75% for the in-situ-micronized drug when the pure drug powder was dispersed without any device. Delivery of the engineered particles via the Spinhaler, the FlowCaps Inhaler and the Cyclohaler increased the FPF from 11 to 46%, 19 to 51%, and 8 to 40%, respectively. PMID- 12637094 TI - A study of the freeze-drying conditions of calixarene based solid lipid nanoparticles. AB - In this note, we report a study of cryoprotectant carbohydrate (glucose, fructose, mannose and maltose) effects on the reconstitution of calixarene based solid lipid nanoparticle (SLN) suspensions after freeze-drying, studied by atomic force microscopy and photon correlation spectroscopy. All carbohydrates tested showed excellent cryoprotection and redispersion properties with the calixarene based SLNs. PMID- 12637095 TI - In vitro evaluation of the viscoelastic properties of chitosan-thioglycolic acid conjugates. AB - The aim of this study was to evaluate the viscoelastic properties of chitosan thioglycolic acid conjugates with different amounts of thiol groups immobilized on the polymer. The modification of chitosan was achieved via the covalent attachment of thioglycolic acid mediated by a carbodiimide. Chitosan-thioglycolic acid conjugates displaying 120, 209 and 439 microM thiol groups per gram of polymer were synthesized. The rheological properties of the three different conjugates were investigated. The elastic properties of the gels were found to increase significantly at pH 5.5. After 6 h the elastic modulus G' of chitosan TGA 120, chitosan-TGA 209 and chitosan-TGA 439 gels increased 7-, 32- and 168 fold, respectively. In parallel the formation of disulfide bonds was observed. Accordingly, proof of principle that chitosan modified by the introduction of thiol groups has in situ gelling properties due to the formation of inter- and intramolecular disulfide bonds at physiological pH values is provided. Based on their in situ gelling properties, chitosan-thioglycolic acid conjugates seem to be very promising new excipients for liquid or semisolid formulations which should stabilize themselves once applied on the site of drug delivery. PMID- 12637096 TI - Complex formation of sericoside with hydrophilic cyclodextrins: improvement of solubility and skin penetration in topical emulsion based formulations. AB - The main objective of this study was to devise novel methods for improving the solubility of the anti-inflammatory triterpenoid sericoside, the main component of Terminalia sericea extract, thus enabling its incorporation into topical formulations. Sericoside was stabilized by complex formation with hydrophilic derivatives of beta- and gamma-cyclodextrins in a molar ratio of 1.0:1.1. The complex of extract and cyclodextrin was equilibrated in water at 25 degrees C for approximately 24 h. The dehydrated complexes of T. sericea extract and cyclodextrin were characterized by differential scanning calorimetry, thermogravimetry analysis and X-ray diffraction. Complex formation with beta cyclodextrin as well as gamma-cyclodextrin derivatives was detectable using these three analytical tools; however, only complexes with gamma-cyclodextrin derivatives showed stability upon storage after incorporation into topical o/w or w/o formulations. Furthermore, a T. sericea extract/gamma-cyclodextrin complex incorporated in an o/w formulation resulted in a 2.6-fold higher percutaneous penetration of sericoside in in vitro excised pig skin as compared to pure T. sericea extract. For the first time, the virtually insoluble anti-inflammatory active sericoside was incorporated into a topical emulsion based formulation in a stable manner, resulting in efficient skin penetration. PMID- 12637097 TI - pH-sensitive polymeric physical-mixture for possible site-specific delivery of ibuprofen. AB - Delivery of drugs to the large bowel has been extensively investigated during the last decade. The aim of this work was to study polymethacrylic acid-co methylmethacrylate substituted with fatty acids (lauric, myristic, palmitic and stearic) at 20% substitution degree (PMA-LAUR20, PMA-MIR20, PMA-PALM20 and PMA STEA20) or 40% substitution degree (PMA-LAUR40, PMA-MIR40, PMA-PALM40 and PMA STEA40) for preparing a pH-sensitive physical mixture for site-specific delivery of ibuprofen chosen as a model drug. The preparation and characterization of the substituted polymers were described. In vitro release studies were conducted at different pH levels (3 h at pH 2.0, 2 h at pH 5.5, 4 h at pH 7.4 and until 24 h at pH 7.0) and phase-solubility diagrams of ibuprofen with the different substituted polymers were obtained at pH 7.0 to obtain information on the influence of amphiphilic polymers in increasing drug solubility and drug availability in the colon. PMID- 12637098 TI - Synthesis, characterization and in vitro release of 5-aminosalicylic acid and 5 acetyl aminosalicylic acid of polyanhydride--P(CBFAS). AB - A novel polyanhydride, poly[(5-carboxybutyl formamide)-2-acetyl salicylic anhydride] (P(CBFAS)), with 5-aminosalicylic acid (5-ASA) incorporated into the polymer backbone was synthesized and characterized by infrared, (1)H-nuclear magnetic resonance, differential scanning calorimetry, vapor pressure osmometry, etc. The polyanhydride was subjected to degradation and simultaneously released 5 ASA and its derivative 5-acetyl aminosalicylic acid (5-acetyl ASA) in vitro under various conditions. The factors influencing the release profiles of 5-ASA and 5 acetyl ASA, including polymer molecular weights, pH value, enzyme and rat gastrointestinal contents, were examined. The results showed that the release rate of 5-ASA and 5-acetyl ASA increases with increasing pH value and with decreasing molecular weights. In PBS (pH 8.0, 37 degrees C) total ASA released was 8.0% for P(CBFAS)(1) (Mn 10770) in 13 h, but only 1.1 and 2.6% at pH 2.0 and 6.5, respectively. Enzymes including pepsin and trypsin, as well as rat gastric and jejunum contents had little effect on the release rate of 5-ASA and 5-acetyl ASA at pH 2.0 and 6.5 (less than 4% in 13 h). However, the release rate of 5-ASA and 5-acetyl ASA was much fast in PBS(pH 8.0) containing 5% of cecal contents, the total ASA released was 13.6% for the polymer in 13 h. Considering the high drug loading of the polymer (50.2% of 5-ASA moieties in the backbones) and the degradation characters, it is possible to reach high local concentration of 5-ASA in the colon site via oral administration. Therefore, P(CBFAS) may be potentially useful in the colon specific delivery of 5-ASA. PMID- 12637099 TI - Stability of fludrocortisone acetate solutions prepared from tablets and powder. AB - To assess the stability of fludrocortisone acetate oral solutions prepared from tablets and powder at three temperatures over a 60-days period. Solutions of fludrocortisone acetate 40 microg/ml were prepared from commercially available 0.05-mg tablets and powder in ethanol 17% v/v. They stored in an amber glass prescription bottles at +4, +23 and +40 degrees C shielded from light. The concentrations of fludrocortisone acetate were determined in duplicate by high performance liquid chromatography at 0, 1, 7, 14, 30, 50 and 60 days. The initial and final pH of solutions were compared. The recovery of fludrocortisone acetate from tablets was determined. The times (t(90)) needed for fludrocortisone acetate to fall to 90% of it's initial concentration were calculated by a linear regression analysis to allow the determination of the expired dates. The recovery of fludrocortisone acetate from tablets was 78 +/- 3%. The t(90) expressed with 95% confidence limits were 2 +/- 1 and 22 +/- 3 days for the solutions prepared from tablets and stored at +23 and +4 degrees C, respectively, whereas t(90) were 11 +/- 2 days and at least 60 days for the solutions prepared with the powder and stored at +23 and +4 degrees C, respectively. No color or odour changes were observed during the study period. The initial pH of the solutions prepared from tablets and powder were 7.7 and 6.9, respectively. No change of pH values was observed at the end of the 60 days. Significant degradation of fludrocortisone acetate occurred in formulations stored at +23 degrees C. Fludrocortisone acetate 40 microg/ml solutions prepared from tablets and powder were stable 19 days and at least 60 days, respectively, when stored at +4 degrees C. The solution prepared from powder is the best in term of stability and final concentration which is independent on the fludrocortisone acetate recovery. PMID- 12637100 TI - Random and systematic measurement uncertainties considered in the evaluation of parameters derived from the course of the tabletting process. AB - The uncertainty of some characteristic parameters describing the course of the tabletting process, namely the area quotient according to Emschermann and Muller, the apparent net work, the slope of the Heckel-plot, and the parameters of the modified Weibull function, was calculated according to the German norm DIN 1319-4 1999. The method allows to consider random and systematic uncertainties in a consistent way as variances of normal and rectangular probability distributions, respectively, or other suitable probability distributions based on Bayesian statistics and the principle of maximum entropy. So, random and systematic uncertainties known from a calibration and validation study of the measurement of force and displacement, and the uncertainty of the true density were included meaningfully into the uncertainty of the resulting tabletting parameters using the propagation of uncertainties according to the Gauss method. The standard uncertainty for the results calculated this way seems to be suitable for a critical evaluation of the tabletting data. PMID- 12637101 TI - Release of small water-soluble drugs from multiblock copolymer microspheres: a feasibility study. AB - Poly(ethylene glycol)-terephthalate/poly(butylene terephthalate) (PEGT/PBT) multiblock copolymer was investigated as a possible matrix for controlled delivery of small water-soluble drugs. Two molecules were selected as sustained release candidates from microspheres: leuprorelin acetate (peptide of Mw = 1270 D) and vitamin B(12) (Mw = 1355 D). First, vitamin B(12)-loaded microspheres were prepared using a double emulsion method and preparation parameters were varied (surfactant in the first emulsion and copolymer composition). The resulting microsphere structure, entrapment efficiency and release rate were evaluated. Vitamin B(12)-loaded microsphere parameters could easily be tailored to achieve specific requirements. The addition of surfactant in the first preparation process led to a significant increase of the microsphere entrapment efficiency, whereas the decrease of the PEGT copolymer content allowed the release rates from microspheres to be precisely decreased. However, leuprorelin acetate-loaded microspheres did not show the same characteristics when prepared with the same parameters, possibly because of a high water solubility discrepancy between the vitamin B(12) and the peptide. This study shows the suitability of PEGT/PBT microspheres as a controlled release system for vitamin B(12), but not for leuprorelin acetate. It also underlines the necessity of tailored development for each individual drug and emphasizes the risk of using model molecules. PMID- 12637102 TI - Preparation of bupivacaine-loaded poly(epsilon-caprolactone) microspheres by spray drying: drug release studies and biocompatibility. AB - Poly(epsilon-caprolactone) microspheres containing bupivacaine were prepared by the spray-drying process. The average size of drug loaded microspheres was less than 3 microm in diameter, and the percentage of entrapment efficiency was 91 +/- 3%. In vitro drug release kinetic in phosphate buffer at 37 degrees C showed a hyperbolic profile, with a burst-effect during the first hour. Subcutaneous injection of bupivacaine-loaded microspheres in the back of rats caused an increase in drug concentration in plasma. Maximum bupivacaine concentration in plasma was 237 +/- 58 ng/ml at 105 h, and drug was detected in plasma for 16 days. The half-life time of the drug was increased by more than 125 times with regard to that of the drug administered in a solution by intraperitoneal injection. After 30 days of injection, a mass formed by microspheres surrounded by a thin fibrous capsule was observed. Small blood vessels and multinucleate foreign body giant cells with macrophagic function around microspheres were detected. After 60 days of injection a subcutaneous mass was also observed, which was formed of more degraded dispersed microspheres in conjunctive tissue, which had a normal structure. Thus, bupivacaine-loaded poly(epsilon-caprolactone) microspheres could be considered as a device to be used in the treatment of severe pain that is not responsive to opioids for example in cancer-related syndromes or in intractable herpetic neuralgia. PMID- 12637103 TI - Preliminary assessment of alginic acid as a factor buffering triethanolamine interacting with artificial skin sebum. AB - Triethanolamine is an ethanolamine used as counter-ion for fatty acid soaps. Interaction between triethanolamine and free fatty acids is suggested to be useful for cleansing sebaceous follicles in acne prevention. This study describes the preliminary assessment of alginic acid as a factor buffering triethanolamine interacting with stearic acid--a compound of artificial skin sebum. Penetration of triethanolamine into artificial sebum, induced by the above mentioned interaction, was measured using a specific optical method. The values of the penetration depth amounted to 0.07-5.74 mm. pH values were measured. The value of pH is reduced from 10.06 for pure 1.49% (w/w) aqueous triethanolamine solution to 6.61 with the increase of the alginic acid to triethanolamine ratio in the preparations. The data of this in vitro research will support further study on other anionic polymers as factors buffering ethanolamines penetrating artificial skin sebum. PMID- 12637104 TI - Transintestinal secretion of ciprofloxacin, grepafloxacin and sparfloxacin: in vitro and in situ inhibition studies. AB - The influence of the secretion process on the absorption of ciprofloxacin, grepafloxacin and sparfloxacin has been evaluated by means of inhibition studies. Two well known P-glycoprotein inhibitors (cyclosporine, verapamil), a mixed inhibitor of P-glycoprotein and the organic cation transporter OCT1 (quinidine) and a well established MRP substrate (p-aminohipuric acid) have been selected in order to distinguish the possible carriers implicated. An in situ rat gut perfusion model and CACO-2 permeability studies are used. Both methods suggest the involvement of several types of efflux transporters for every fluoroquinolone. The relevance of the secretory pathway depends on the intrinsic permeability of the quinolone. The in vitro model seems to be more suitable for discriminating mechanisms underlying the absorption process, while in situ studies are less sensitive to inhibition studies. PMID- 12637105 TI - Phase solubility studies of pure (-)-alpha-bisabolol and camomile essential oil with beta-cyclodextrin. AB - (-)-Alpha-bisabolol was found to form an inclusion complex with beta-cyclodextrin (beta-CD) in solution as well as in the solid state. To investigate molecular associations of beta-CD with pure (-)-alpha-bisabolol or (-)-alpha-bisabolol as a component of camomile essential oil, phase solubility studies were undertaken. A B(s) type solubility with an apparent complex constant of 273 M(-1) for the pure (-)-alpha-bisabolol and 304 M(-1) for (-)-alpha-bisabolol as a constituent of the essential oil were obtained. The two curves in the phase solubility diagram reach their plateau at different concentrations of (-)-alpha-bisabolol, 7.04 x 10(-4) M for the pure substance and 2.88 x 10(-4) M for the substance as a component of the essential oil. Although the shapes of the curves are almost similar, the intrinsic solubility's of pure (-)-alpha-bisabolol (4.85 x 10(-4) M) and (-) alpha-bisabolol as a component of the essential oil (1.82 x 10(-4) M) differ significantly. An inclusion complex having a stoichiometric composition of 2:1 (beta-CD: drug) was obtained. A mechanism of complexation has been proposed on the basis of the stability constant calculated from phase solubility data and the stoichiometric ratio of the solid state complexation. PMID- 12637110 TI - Effect of daily and intermittent use of ibandronate on bone mass and bone turnover in postmenopausal osteoporosis: a review of three phase II studies. AB - BACKGROUND: Oral bisphosphonates are well established for the treatment and prevention of postmenopausal osteoporosis; however, they are poorly absorbed from the gastrointestinal (GI) tract and have been associated with GI adverse events. Thus, current dosing guidelines recommend that the patient not eat or lie down for at least 30 minutes after taking oral bisphosphonates, a requirement that is inconvenient and may be associated with reduced compliance. The drawbacks of these dosing requirements may be overcome either by reducing dosing frequency or by using alternative routes of administration. OBJECTIVE: Ibandronate is a potent nitrogen-containing bisphosphonate that can be given orally or IV, daily or intermittently, with a between-dose interval of up to 3 months. This article presents the results of published Phase II trials of the efficacy and safety profile of oral and IV ibandronate administered daily or intermittently to postmenopausal women with low bone mass. METHODS: MEDLINE was searched through January 2002 to identify all published Phase II clinical studies of oral and IV ibandronate in the treatment of post-menopausal osteoporosis. RESULTS: In the 3 Phase II studies identified, marked reductions in biochemical markers of bone resorption (50%-70%) and bone formation (40%-50%) were seen to a similar and statistically significant extent with oral ibandronate 2.5 mg/d (P<0.001), oral ibandronate 20 mg QOD given for 12 doses at the start of each 3-monthly period (P<0.001), and injections of ibandronate 2 mg IV given every 3 months (P<0.01). All treatment regimens produced comparable significant increases in bone mineral density at the lumbar spine (P<0.01) and hip (P<0.05). Ibandronate was well tolerated when administered both orally and as an IV injection. CONCLUSIONS: In these Phase II studies, oral or IV ibandronate, administered continuously or intermittently, reduced markers of bone turnover, significantly increased bone mineral density, and was well tolerated in the treatment of osteoporosis in postmenopausal women. The data from these studies provided the rationale for further investigation of ibandronate in larger longer-term Phase III studies evaluating its potential as an efficacious and flexible alternative to existing bisphosphonate regimens. PMID- 12637109 TI - The impact of nonsteroidal anti-inflammatory drugs on blood pressure, with an emphasis on newer agents. AB - BACKGROUND: Both nonselective and selective nonsteroidal anti-inflammatory drugs (NSAIDs) have been reported to increase blood pressure (BP). OBJECTIVE: This review summarizes the most recent data regarding the BP changes that may occur with routine dosing of nonselective or selective NSAIDs, with a focus on the latter. The review includes both clinical trials and studies having more naturalistic designs. METHODS: Relevant articles were identified through a search of MEDLINE emphasizing NSAIDs and hypertension. RESULTS: It is well established that NSAIDs increase BP in certain patients. Patients with existing hypertension seem to be at greatest risk for this adverse event. There seem to be differences between nonselective NSAIDs with regard to their effect on BP; indomethacin, naproxen, and piroxicam have been associated with clinically significant changes in BP. Most of the available evidence concerning the selective NSAIDs suggests that rofecoxib is more likely than celecoxib to raise systolic BP. Based on the results of trials in the treatment of BP, increases in systolic BP produced by these NSAIDs may be associated with significant morbidity and mortality. The adverse effects of NSAIDs on BP may be related to the specific drug or to the dose or duration of therapy, as well as to patient-specific characteristics. CONCLUSIONS: Clinically significant BP changes may occur after treatment with either nonselective or selective NSAIDs. The observed BP changes are not the same for all products, however. PMID- 12637111 TI - Effect of amlodipine on systolic blood pressure. AB - BACKGROUND: Systolic hypertension is the most common form of hypertension, particularly in people aged >60 years. Caused by decreased compliance of large arteries, systolic hypertension is an independent risk factor for cardiovascular disease. Recent studies have demonstrated that it is more important to control systolic blood pressure (SBP) than diastolic blood pressure (DBP). OBJECTIVE: The objective of this study was to perform a systematic literature review to examine the effectiveness of amlodipine in lowering SBP in a variety of patient subgroups and clinical settings. METHODS: The literature review methodology included identifying, selecting, appraising, extracting, and synthesizing primary research studies. Following an a priori protocol, published literature was searched from 1980 to 2001 using 3 electronic databases. A manual review of the reference lists of recent review articles and all accepted studies was performed. Parallel-group, randomized, controlled trials that included at least 10 adults with baseline hypertension (SBP>or=140 mm Hg, DBP>or=90 mm Hg, or both), included at least 1 arm randomized to initial treatment with amlodipine monotherapy, had a minimum treatment duration of 8 weeks, and reported baseline and end-point blood pressure were included. RESULTS: Of 696 citations identified, 85 primary studies met all inclusion criteria. Comparable treatment arms were pooled, and weightd mean SBP was calculated. In the amlodipine monotherapy arms, which included >5000 patients, SBP decreased by a mean of 17.5 mm Hg from baseline. The effect of amlodipine in reducing SBP was greater in elderly patients (age>or=60 years) and patients with author-defined isolated systolic hypertension. The dose was titrated to achieve the target blood pressure in 73 of 89 amlodipine treatment arms, whereas 16 treatment arms reported fixed doses. The median daily dose was 5 mg (range, 1.25-15 mg) in both the fixed-dose and dose-titration groups. CONCLUSIONS: In this review of the published literature, amlodipine monotherapy was effective in reducing SBP. Antihypertensive agents such as amlodipine warrant consideration for the management of patients with inadequately controlled SBP. PMID- 12637112 TI - Weight management and current options in pharmacotherapy: orlistat and sibutramine. AB - BACKGROUND: Chronic obesity is associated with various cardiovascular disorders, including diabetes, dyslipidemia, and hypertension. Pharmacotherapy with antiobesity agents is an important management strategy in conjunction with lifestyle interventions. OBJECTIVE: This article describes the pharmacologic management of obesity, concentrating on orlistat and sibutramine. METHODS: Relevant articles were identified through a MEDLINE search (1966-February 2002) using the terms obesity, overweight, weight loss, antiobesity drugs, orlistat, and sibutramine. The search for efficacy trials was limited to randomized controlled studies of >6 months' duration. Also included in the review were relevant references cited in the bibliographies of identified articles, news reports, and the authors' own data. RESULTS: Orlistat reduces fat absorption by inhibiting gastrointestinal lipases. In randomized, controlled trials of up to 2 years' duration, orlistat plus a hypocaloric diet produced significantly greater weight loss than placebo (P < 0.001). In the maintenance phase, patients taking orlistat had less weight regain than did placebo recipients. The weight reduction with orlistat was also associated with a significant improvement in control of cardiovascular risk factors (P < 0.05). Unlike orlistat, sibutramine works by suppressing appetite; its efficacy, however, was similar to that of orlistat in the identified clinical trials. Orlistat was associated primarily with gastrointestinal side effects. Use of orlistat was associated with minimal drug interactions, except with cyclosporine, with which it should not be taken. Sibutramine was also well tolerated, although it may cause dry mouth, anorexia, and insomnia, and should be used with caution in patients at risk for cardiovascular disease. CONCLUSIONS: Orlistat and sibutramine demonstrated a favorable efficacy and safety profile in randomized controlled trials. Current evidence supports their use as adjuncts to lifestyle modifications in the treatment of obesity. PMID- 12637113 TI - Gabapentin dosing for neuropathic pain: evidence from randomized, placebo controlled clinical trials. AB - BACKGROUND: Pain is one of the most common reasons for seeking medical attention, and neuropathic pain is among the most common types of pain. Despite its prevalence, neuropathic pain is often underrecognized and inadequately treated. Many cases are refractory to the medications traditionally used for pain, such as nonsteroidal anti-inflammatory drugs. Tricyclic antidepressants are considered first-line agents for neuropathic pain, but their use is limited by unwanted side effects and a risk of cardiovascular mortality. OBJECTIVES: The goals of this article were to review data on the efficacy and tolerability of gabapentin in the treatment of neuropathic pain in adults and to determine the optimal dosing schedule. METHODS: Randomized controlled studies of gabapentin for neuropathic pain were identified through a search of PubMed and MEDLINE from 1966 to the present using the search terms gabapentin, randomized, placebo, and pain. Abstracts of identified articles were screened for study size (>100 patients per treatment arm) and use of appropriate efficacy measures. A separate review based on information provided by the manufacturer of gabapentinaand clinical trial Web sites was conducted to ascertain whether there had been any other relevant industry- or government-sponsored trials. The manufacturer provided additional unpublshed study data. RESULTS: Data from 5 randomized, placebo-controlled trials were included in the review, 1 of which has not yet been published. Gabapentin was effective in the treatment of painful diabetic neuropathy, postherpetic neuralgia, and other neuropathic pain syndromes. It relieved symptoms of allodynia, burning pain, shooting pain, and hyperesthesia. Adverse effects were typically mild to moderate and usually subsided within approximately 10 days from the initiation of treatment. Based on available data, it appears that treatment should be started at a dose of 900 mg/d (300 mg/d on day 1, 600 mg/d on day 2, and 900 mg/d on day 3). Additional titration to 1800 mg/d is recommended for greater efficacy. Doses up to 3600 mg/d may be needed in some patients. The effective dose should be individualized according to patient response and tolerability. CONCLUSION: At doses of 1800 to 3600 mg/d, gabapentin was effective and well tolerated in the treatment of adults with neuropathic pain. PMID- 12637114 TI - An examination of the results of the EVIDENCE, INCOMIN, and phase III studies of interferon beta products in the treatment of multiple sclerosis. AB - BACKGROUND: Three interferon (IFN) beta products are currently available for the treatment of relapsing multiple sclerosis (MS). Each of these agents showed effectiveness in the treatment of MS in the respective randomized, double-blind, placebocontrolled Phase III trials. However, there have been no randomized, double-blind, placebo-controlled trials directly comparing the efficacy and safety of these formulations. OBJECTIVE: The objective of this article was to compare the results of available comparative studies with the results of the pivotal Phase III trials of each IFN beta product. METHODS: BIOSIS, Current Contents/Clinical Medicine, and MEDLINE were searched for English-language articles published from 1996 to the present comparing the efficacy and safety of IFN beta formulations in the treatment of MS. Search terms included interferon beta 1a, interferon beta 1b, and multiple sclerosis. Articles or abstracts that reported the results of Phase III trials or studies directly comparing IFN beta formulations in the treatment of relapsing or relapsing-remitting MS were included in the review. RESULTS: Seven head-to-head studies were identified that directly compared the efficacy of IFN beta products in the treatment of MS. Two of these studies- INCOMIN (Independent Comparison of Interferon) and EVIDENCE (Evidence for Interferon Dose-Effect: European-North American Comparative Efficacy)- found significant differences in clinical efficacy between IFN beta products, whereas the remaining studies showed equal clinical efficacy between products. CONCLUSION: Inconsistencies within and between the results of the reviewed studies suggest that clinicians should use caution in interpreting the findings of the INCOMIN and EVIDENCE comparative trials. PMID- 12637115 TI - A 52-week, multicenter, randomized, parallel-group, double-blind, double-dummy study to assess the efficacy of atorvastatin and simvastatin in reaching low density lipoprotein cholesterol and triglyceride targets: the treat-to-target (3T) study. AB - BACKGROUND: Guidelines for the prevention of coronary heart disease call for low density lipoprotein cholesterol (LDL-C) reduction as the primary target of treatment and reduction of triglycerides (TG) as an additional target. OBJECTIVE: The purpose of this study was to investigate the ability of atorvastatin and simvastatin to reduce LDL-C and TG concentrations and to meet 3 target lipid levels: LDL-C or=4.0 mmol/L (>or=155 mg/dL), were randomized in a 1:1 ratio to receive once-daily oral treatment with 20 mg atorvastatin or 20 mg simvastatin. Fasting (12-hour) blood samples for the estimation of lipid levels and clinical laboratory values were collected after 4, 8, 12, 26, and 52 weeks. The dose was doubled after 12 weeks if the target National Cholesterol Education Program level of LDL-C (or=1 cardiorenal risk factor, and approximately one third had hypertension. Cardiorenal risk factors were not related to physicians' prescribing of celecoxib or rofecoxib, but the presence of any cardiorenal risk factor was associated with an increase in the use of COX-2-selective inhibitors compared with other NSAIDs, from 12% for cerebrovascular disease (point estimate, 1.124; P<0.001) to 74% for chronic renal failure/nephritis (point estimate, 1.738; P=0.025). RA and advanced age were associated with the use of celecoxib rather than rofecoxib. CONCLUSIONS: The prevalence of cardiorenal risk factors was found to be similar in patients prescribed celecoxib or rofecoxib for arthritis. Patients with these risk factors were more likely to receive a COX-2-selective inhibitor than other NSAIDs. PMID- 12637119 TI - Cognitive improvement in mild to moderate Alzheimer's dementia after treatment with the acetylcholine precursor choline alfoscerate: a multicenter, double blind, randomized, placebo-controlled trial. AB - BACKGROUND: Parallel with the development of hypotheses regarding cholinergic involvement in geriatric memory dysfunction, the first attempts to treat patients with Alzheimer's disease (AD) involved the cholinergic-precursor loading approach. Despite encouraging early results, well-controlled clinical trials did not confirm a clinical utility of cholinergic precursors such as choline and lecithin (phosphatidylcholine) in AD. OBJECTIVE: This study assessed the efficacy and tolerability of the cholinergic precursor choline alfoscerate (CA) in the treatment of cognitive impairment due to mild to moderate AD. METHODS: In this multicenter, double-blind, randomized, placebo-controlled trial, patients affected by mild to moderate dementia of the Alzheimer type were treated with CA (400-mg capsules) or placebo capsules, 3 times daily, for 180 days. Efficacy outcome measures that were assessed at the beginning of the investigation and after 90 and 180 days of treatment included scores of the Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-Cog), the Mini-Mental State Examination (MMSE), the Global Deterioration Scale (GDS), the Alzheimer's Disease Assessment Scale-Behavioral Subscale (ADAS-Behav), all items of the Alzheimer's Disease Assessment Scale (ADAS-Total), and the Clinical Global Impression (CGI) scale. The Global Improvement Scale (GIS) score was assessed after 90 and 180 days of treatment. RESULTS: A total of 261 patients (132 in the CA group, 129 in the placebo group) were enrolled in the study. The mean (SD) age in the CA group was 72.2(7.5) years (range, 60-80 years), and in the placebo group it was 71.7 (7.4) years(range, 60-80 years). The CA group comprised 105 women and 27 men; the placebo group, 94 women and 35 men. The mean decrease in ADAS-Cog score in patients treated with CA was 2.42 points after 90 days of treatment and 3.20 points at the end of the study (day 180) (P < 0.001 vs baseline for both), whereas in patients receiving placebo the mean increase in ADAS-Cog score was 0.36 point <1 after 90 days of treatment and 2.90 points after 180 days of treatment(P < 0.001 vs baseline). In the CA group, all other assessed parameters (MMSE,GDS, ADAS-Behav, ADAS-Total, and CGI) consistently improved after 90 and 180 days versus baseline, whereas in the placebo group they remained unchanged or worsened. Statistically significant differences were observed between treatments after 90 and 180 days in ADAS-Cog, MMSE, GDS, ADAS-Total, and CGI scores and after 180 days of treatment in ADAS-Behav and GIS scores. CONCLUSION: The results of this study suggest the clinical usefulness and tolerability of CA in the treatment of the cognitive symptoms of dementia disorders of the Alzheimer type. PMID- 12637118 TI - Activity of nine oral agents against gram-positive and gram-negative bacteria encountered in community-acquired infections: use of pharmacokinetic/pharmacodynamic breakpoints in the comparative assessment of beta lactam and macrolide antimicrobial agents. AB - BACKGROUND: The application of pharmacokinetic (PK) and pharmacodynamic (PD) data in conjunction with minimum inhibitory concentrations (MICs) of antibacterial agents has been shown to allow for improved selection and appropriate dosing of antimicrobial agents for specific infections, increasing the likelihood of bacteriologic cure and, through this, reducing the risk for the development of resistant organisms. OBJECTIVES: This study was undertaken to provide data on current levels of resistance among common community-acquired bacterial species to 7 betalactam antimicrobial agents (including the combination product amoxicillin/clavulanate), azithromycin, and clarithromycin, determined through application of the PK/PD breakpoints based on time-above-MIC for the beta-lactams and the nonazalide macrolide clarithromycin, and on 24-hour serum area under the curve divided by MIC for the azalide macrolide azithromycin. METHODS: The antimicrobial products tested were amoxicillin/clavylanate, cefpodoxime, cefdinir, cefditoren, cefprozil, cefuroxime, cefixime, azithromycin, and clarithromycin. The bacterial species comprised 70 penicillin-susceptible, 68 penicillin-intermediate, and 69 penicillin-resistant strains of Streptococcus pneumoniae; 46 beta-lactamase-positive and 54 beta-lactamase-negative strains of Haemophilus influenzae; 49 strains of Moraxella catarrhalis; and 100 methicillin sensitive strains of Staphylococcus aureus (MSSA). Strains were isolated from clinical specimens obtained from outpatient-acquired infections in 1 clinical center in the Northeast and 1 in the north-central area of the United States within the past 2 years. National Committee for Clinical Laboratory Standards microdilution MIC methodology was used. PK/PD breakpoints were obtained from previously published studies and were based on blood values. RESULTS: Amoxicillin/clavulanate was the product to which the greatest percentage of susceptible, intermediate, and resistant strains of pneumococci were sensitive at the PK/PD breakpoint, followed by cefditoren, cefpodoxime, cefuroxime, cefdinir, and cefprozil. None of the cephalosporins were active against penicillin resistant pneumococci. Cefditoren and cefpodozime were the agents to which the greatest percentage of beta-lactamase-positive and beta-lactamase-negative strains of H influenzae were sensitive, followed by amoxicillin/clavulanate, cefdinir, and cefuroxime. Cefprozil was inactive against H influenzae. All of the beta-lactam products were active against M catarrhalis. All but cefpodoxime, cefditoren, and cefixime were active against MSSA. CONCLUSIONS: In this study, based on PK/PD breakpoints, amoxicillin/clavulanate had the best overall activity of the 9 antimicrobial products tested. Cefpodoxime and cefditoren were active against >or=90% of strains of penicillin-susceptible and penicillin-intermediate pneumococci, H influenzae, and M catarrhalis. The macrolides azithromycin and clarithromycin were active against penicillin-susceptible and penicillin intermediate pneumococci and M catarrhalis; they were inactive against H influenzae and penicillin-resistant pneumococci. PMID- 12637120 TI - Efficacy and safety profile of glimepiride in Mexican American Patients with type 2 diabetes mellitus: a randomized, placebo-controlled study. AB - BACKGROUND: Mexican Americans, the fastest growing ethnic group in the United States, have a 2- to 3-fold higher prevalence of type 2 diabetes mellitus relative to the non-Hispanic white population. It is estimated that 10% of Mexican Americans >or=20 years of age have diabetes. OBJECTIVE: The goal of this study was to evaluate the efficacy and safety of glimepiride, a long-acting sulfonylurea, as an adjunct to diet/exercise in Mexican Americans with type 2 diabetes mellitus. METHODS: This was a multicenter, randomized, double-blind, placebo-controlled study. Mexican Americans with uncontrolled type 2 diabetes, defined as a fasting plasma glucose (FPG) level between 120 mg/dL and 225 mg/dL and glycated hemoglobin (HbA(1c)) values between 8.0% and 10.5%, after >or=3 months of diet/exercise were enrolled. Patients were randomized in a 2:1 ratio (using the lowest available treatment assignment number when eligibility was established) to receive 14 weeks of glimepiride or matching placebo once daily with continued diet/exercise. The starting glimepiride dose was 1 mg, with titration to 2 mg and 4 mg for FPG levels >120 mg/dL. The primary efficacy variable was change in HbA(1c) from baseline to study end point. Secondary efficacy variables were HbA(1c) response (rated as excellent, good, or marginal) and changes in FPG, fasting insulin, fibrinogen, and plasminogen activator inhibitor-1 (PAI-1) levels from baseline to study end point. The tolerability of glimepiride in this study population was determined by evaluating adverse events, hypoglycemic episodes, and physical examination as well as laboratory findings. All analyses were performed on an intent-to-treat basis. A per-protocol analysis also was conducted to support the primary efficacy analysis. RESULTS: Seventy patients were randomized to treatment with glimepiride (n = 48) or placebo ((n = 22). The glimepiride and placebo groups were similar with respect to mean (SE) age (48.4 [11.7] and 50.7 [10.0] years, respectively) and sex (56.3% [27/48] and 50.0% [11/22] were male, respectively). However, the glimepiride group had a higher mean body weight (83.3 [17.0] vs 76.3 [18.5] kg) and a significantly higher mean fasting insulin level (23.8 [17.7] vs 17.8 [19.7] microU/mL; P = 0.031). The mean (SE) HbA(1c) values at study end point were 7.8% (0.2%) and 9.9% (0.7%) in patients receiving glimepiride and placebo, respectively. The adjusted mean difference in HbA(1c) reduction from baseline to end point was statistically significant in favor of glimepiride (-1.8% [0.4%]; P < 0.001). More pronounced HbA(1c) impairment at baseline was associated with greater glimepiride-placebo differences in HbA(1c) reduction. Glimepiride-treated patients also achieved a significantly greater improvement in FPG, with an adjusted mean (SE) treatment difference of -46.7 (16.7) mg/dL (P = 0.007). Glimepiride did not appear to affect fibrinogen and PAI-1 levels but was associated with significantly greater mean increases in fasting insulin (10.2 vs -2.1 microU/mL; P = 0.002) and body weight (2.3 vs 2.1 kg; P < 0.001) compared with placebo. Glimepiride was well tolerated, with an adverse-event profile similar to that of placebo. CONCLUSIONS: These results indicate that once-daily glimepiride plus diet/exercise was effective in Mexican Americans with type 2 diabetes whose disease was inadequately controlled with diet/exercise alone. It appeared to be well tolerated in the population studied. More weight gain was seen with glimepiride compared with placebo. Given the high prevalence of type 2 diabetes among Mexican Americans, further clinical studies of glimepiride and other glucose-lowering therapies are needed in this ethnic subset. PMID- 12637121 TI - An open-label, nonrandomized, single-center, prospective extension, clinical trial of booster dose schedules to assess the safety profile and immunogenicity of recombinant outer-surface protein A (OspA) Lyme disease vaccine. AB - BACKGROUND: An efficacy trial of an outer-surface protein A (OspA) Lyme disease vaccine demonstrated tolerability and efficacy against laboratory-confirmed Lyme disease after a primary series of 3 doses at 0, 1, and 12 months. OBJECTIVES: This extension of the efficacy study assessed the immunogenicity and tolerability of booster vaccinations administered at 24 and/or 36 months after the first vaccination. METHODS: This open-label, nonrandomized, single-center, prospective extension, clinical trial was conducted in the general community in New Haven, Connecticut, where Lyme disease is endemic. Blood samples (to determine anti-OspA titer) were collected before administration of the booster doses at months 24 and 36, and at 1 and 12 months after each booster dose was administered. Immune response was assessed via total immunoglobulin G (IgG) anti-OspA antibody titers and the proportion of subjects with titers >or=1400 EL.U/mL. Adverse events (AEs) were recorded by the study volunteers on diary cards. RESULTS: A total of 318 volunteers (173 women and 145 men) received at least 1 booster dose of Lyme disease vaccine, administered at 12 or 24 months after the third vaccination of the primary series (months 24 and 36, in relation to the primary series). Eighty eight subjects of those who received a month-24 booster received a second booster dose at month 36 (12 months after the first booster). Overall, the mean age of the volunteers was 55 years (range, 19 to 73 years). The demographic characteristics of the groups were similar. Most AEs were limited induration and were rated by investigators and subjects as mild to moderate in severity. Administration of I or 2 booster doses did not elicit any patterns of AEs different from those reported in the efficacy trial. After the first booster dose, all volunteers had an anamnestic response and positive test results for total IgG antibody. Geometric mean titers increased at least 12-fold 1 month after the first booster dose at month 24 or 36. More than 96% of volunteers had titers>1400 EL.U/mL and 100% had titers >400 EL.U/mL (minimum seroprotective level) 1 month after the booster dose at month 24 or 36. CONCLUSIONS: All booster doses were well tolerated, and the incidence of AEs did not increase after the second booster dose. The immune response generated after the 3-dose primary series waned; booster doses administered at 12 and/or 24 months after the primary series increased antibody levels above seroprotective levels. PMID- 12637122 TI - Open-label, sequential comparison of eptifibatide with abciximab for patients undergoing percutaneous coronary intervention. AB - BACKGROUND: Platelet glycoprotein IIb/IIIa antagonists reduce complications following percutaneous coronary intervention (PCI). There are limited data comparing different agents. OBJECTIVE: The purpose of this study was to compare in-hospital and 30-day outcomes in 2 sequential cohorts of consecutive patients undergoing PCI at our institution who received abciximab or eptifibatide. METHODS: The first cohort included patients who received abciximab between September 1, 1998, and January 9, 1999, and the second included patients who were treated with eptifibatide between January 11 and April 27, 1999. Per formulary decision, during the latter period, patients with renal insufficiency continued to be treated with abciximab and were ineligible for therapy with eptifibatide. Major adverse cardiac events (MACEs) were evaluated by one or more of the authors and compared. RESULTS: A total of 319 patients were treated with abciximab and 301 with eptifibatide. There were no differences in baseline characteristics between the 2 groups, with the exception of a significantly higher proportion of patients with chronic renal insufficiency in the abciximab-treated group (4% vs 0% with eptifibatide; P = 0.03) The majority of interventions were performed for an acute coronary syndrome. Procedural success was 97% in both groups. Eptifibatide patients were treated for a mean 20.4 (5.2) hours, with 10.1% receiving the drug before the procedure. There were no differences in overall or individual MACEs in hospital or at 30 days. CONCLUSION: Our data suggest similar in-hospital and 30-day outcomes for abciximab- and eptifibatide-treated patients undergoing PCI. PMID- 12637124 TI - The brief history and uncertain future of prescription drug benefits. PMID- 12637123 TI - An open-label study to assess changes in efficacy and satisfaction with migraine care when patients have access to multiple sumatriptan succinate formulations. AB - BACKGROUND: Because a patient's migraines often differ in duration, intensity, and accompanying symptoms, as well as the conditions and circumstances at the time of the headache, the mode for treatment also may change. OBJECTIVE: The goal of this study was to determine whether migraine management is improved by providing 3 formulations of sumatriptan succinate to patients, together with education to assist them in selecting the most appropriate formulation for specific attacks. METHODS: This was an open-label study conducted in 3 family practice settings. Patients were recruited who had at least a 1-year history of migraine meeting International Headache Society criteria and experienced 2 to 6 attacks per month within the previous 3 months. Patients received instructions on oral, intranasal, and subcutaneous (SC) sumatriptan and were provided with all 3 formulations to treat 6 headaches. Migraine features, formulation used, reason for selecting specific formulation, migraine symptom relief, and use of follow-up doses were recorded in diaries. At follow-up, patients completed a questionnaire assessing satisfaction with access to multiple formulations. RESULTS: Of the 33 enrolled patients (26 women, 7 men; mean age, 38.5 years [range, 23-54 years]), 25 (75.8%) completed all visits. Of 149 headaches treated, 39 (26.2%) were mild at onset, 70 (47.0%) were moderate, and 40 (26.8%) were severe. Eighty (53.7%) headaches were treated with tablets, 35 (23.5%) with nasal spray, and 34 (22.8%) with SC injection. Primary reasons for selecting specific formulations included "fewer side effects" for tablets, "convenience" for nasal spray, and "quick onset of action" for SC injection. Twenty-one (84.0%) patients reported being either very satisfied or satisfied with their ability to manage their headaches. Physicians reported that 18 of 24 (75.0%) patients had an improved attitude toward managing their headaches. All formulations were well tolerated. Eight (32.0%) patients reported adverse events, the 2 most common being chest pressure and fatigue. CONCLUSION: The patients in this study reported greater satisfaction with migraine management when given access to multiple sumatriptan formulations and education regarding their appropriate use. PMID- 12637125 TI - Approaches to pharmacy benefit management and the impact of consumer cost sharing. AB - BACKGROUND: Numerous mechanisms have been introduced to deliver prescription drug benefits while controlling pharmaceutical costs. An understanding of the most prominent mechanisms of benefit management is an important step in determining the most effective approach to take in future years. OBJECTIVES: The aims of this review were to illustrate the mechanisms by which managed care has attempted to efficiently and equitably deliver pharmacy benefits and to discuss the impact of such programs, including consumer cost sharing. METHODS: A review of the literature was conducted using the PreMedline and MEDLINE databases from the years 1966 to 2002, reference lists from relevant articles, and online sources, including news releases, conference materials, and pharmacy benefit management reports. RESULTS: Numerous pharmacy benefit management tools and their impact on utilization, expenditures, and health outcomes are reviewed, including disease state management; utilization management (ie, quantity limitations and prior authorization); drug utilization review; formulary management (ie, open and closed); delivery systems (ie, retail and mail order); and mechanisms for implementing consumer cost sharing (ie, generic incentives, multitiered copayments, and co-insurance). Although there is some evidence to suggest that certain benefit management tools have been successful in reducing health plan expenditures, a more thorough investigation of their potential unintended consequences is needed. CONCLUSIONS: Implementing adequate levels of consumer cost sharing is necessary if employers and health plans are to continue offering prescription drug benefits. It is important to remember, however, that quality health care cannot be forfeited for the sake of short-term cost savings. PMID- 12637126 TI - Physician gender and changes in drug prescribing after the implementation of reference pricing in British Columbia. AB - BACKGROUND: Gender-specific attitudes and communication styles are known to influence both the content and outcome of medical visits. Therefore, gender specific differences in response to cost containment may also occur. OBJECTIVE: The purpose of this study was to assess the effect of physician gender on changes in prescribing patterns of angiotensin-converting enzyme (ACE) inhibitors after the implementation of reference pricing for prescription drugs in British Columbia, Canada. METHODS: Reference pricing is a cost-sharing policy by which use of high-priced medication requires out-of-pocket payment of the price difference between the cost-sharing drug and a lower-cost drug within the same class. In British Columbia, reference pricing for ACE inhibitors was introduced on January 1, 1997. Analysis was carried out on linked pharmacy and medical service claims data on 927 female and 2922 male physicians treating 47,680 Pharmacare Plan A enrollees who were aged >-65 years and were prescribed a high priced ACE inhibitors before the implementation of reference pricing. RESULTS: Female physicians (24.1% of all physicians) were younger, treated more female patients, had patients with fewer chronic illnesses, and worked more often as general practitioners than did male physicians. The patients of female physicians were more likely to receive a written physician-requested exemption from copayment, according to a multivariate logistic regression analysis (odds ratio [OR], 1.25; 95% CI, 1.04-1.50). Data suggested that patients of female physicians were more likely to stop antihypertensive drug therapy (OR, 1.43; 95% CI, 0.96 2.13); however, this was independent of the new copayment policy. CONCLUSIONS: The results provide empirical evidence that physician gender is associated with slightly different patient management strategies regarding physician-requested exemptions after the start of a new drug cost-sharing policy. However, these differences are unlikely to have meaningful clinical or economic consequences. PMID- 12637127 TI - An economic assessment of inhaled formoterol dry powder versus ipratropium bromide pressurized metered dose inhaler in the treatment of chronic obstructive pulmonary disease. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is debilitating and costly to manage. A recent clinical trial concluded that formoterol, a long acting beta(2)-adrenoceptor agonist, was more clinically effective than inpratropium bromide in the management of COPD. OBJECTIVE: The aim of this study was to perform an economic assessment of the management of COPD with formeterol versus ipratropium bromide. METHODS: Assessment were based on the results of a previously published 12-week, multicenter, double-masked, randomized, parallel group, placebo-controlled clinical trial comparing inhaled formoterol dry powder 12 and 24 microg BID with ipratropium bromide 40 microg QID pressurized metered dose inhaler and with placebo in 780 COPD patients. Treatment costs for study drugs and rescue medications were estimated from resource utilization and average wholesale prices. Costs were assessed with respect to forced expiratory lung volume in 1 second (FEV(1)) and patient-reported quality of life (QOL) as assessed via the St. George's Respiratory Questionnaire. Cost-effectiveness ratios were calculated for each treatment arm and incremental cost-effectiveness ratios (ICERs) were calculated for each treatment relative to the next best alternative. Economic efficiency frontiers were identified. Sensitivity analysis was conducted. RESULTS: Cost analysis with respect to FEV(1) revealed an economic efficiency frontier formed by placebo, ipratropium bromide 40 microg, and formoterol 12 microg at their respective FEV(1) levels, with cost-effectiveness ratios of $30.18, $53.50, and $142.04, respectively, per FEV(1). The ICER for ipratropium over placebo was $273.03; for formoterol 12 microg over ipratropium, $1611.32. Cost analysis with respect to QOL showed an economic efficiency frontier formed by placebo and formoterol 12 microg at their respective QOL outcomes, with cost-effectiveness ratios of $25.96 and $32.56, respectively, per QOL score change. The cost-effectiveness ratio for ipratropium was $69.40,which was not on the QOL economic efficiency frontier. The ICER for formoterol 12 microg over placebo was $34.51 per QOL score point. CONCLUSIONS: Formoterol 12 microg provided greater QOL outcome than ipratropium bromide at an additional cost of $554.28 per year. Further research would be necessary to assess whether the differences in outcomes, particularly QOL, observed in the short term with formoterol would lead to favorable long-term health and economic outcomes. PMID- 12637128 TI - Quality-of-life assessment in patients with hyperhidrosis before and after treatment with botulinum toxin: results of an open-label study. AB - BACKGROUND: Patients with hyperhidrosis, a disorder characterized by increased sweat production, experience substantial functional and emotional problems. Botulinum toxin type A (BTX-A) has been shown to be useful in the treatment of hyperhidrosis; however, few studies have considered the effects of treatment on patients' quality of life (QOL). OBJECTIVES: The objectives of this study were to assess QOL in patients with focal hyperhidrosis; to investigate whether the impairment in QOL in these patients is related to the type of hyperhidrosis or the number of sites involved; and to compare the changes in QOL and the response to BTX-A treatment in patients with axillary and palmar hyperhidrosis. METHODS: Patients with focal primary hyperhidrosis of the axillae, palms, and soles who had experienced decreased QOL and whose condition had not responded to conventional topical and physical therapies were included in this open-label study. Patients completed a self-administered Dermatology Life Quality Index (DLQI) questionnaire before and 2 weeks after treatment with BTX-A. RESULTS: All 41 patients had experienced a decrease in QOL as measured by the DLQI. The impairement in QOL was not dependent on the number or types of sites involved. Treatment with BTX-A led to improvement in QOL in all patients, with the median DLQI score decreasing (ie, improving) significantly from pretreatment level (P < 0.001). The improvement in QOL and response to treatment were similar in patients with axillary and palmar hyperhidrosis. CONCLUSIONS: Further studies with a longer follow-up period are needed to assess the long-term effects of BTX-A; however, preliminary data from the present study suggest that BTX-A improves QOL in patients with focal hyperhidrosis, independent of the presenting clinical picture. PMID- 12637129 TI - A 3-stage model for assessing the probable economic effects of direct-to-consumer advertising of pharmaceuticals. AB - BACKGROUND: The pharmaceutical industry employs a variety of marketing strategies that have previously been directed primarily toward physicians. However, mass media direct-to-consumer (DTC) advertising of prescription drugs has emerged as a ubiquitous promotional strategy. OBJECTIVE: This article explores the economics of DTC advertising in greater depth than has been done in the past by using a 3 stage economic model to assess the pertinent literature and to show the probable effects of DTC advertising in the United States. METHODS: Economics literature on the subject was searched using the Journal of Economic Literature. Health services literature was searched using computer callback devices. RESULTS: Spending on DTC advertising in the United States increased from $17 million in 1985 to $2.5 billion in 2000. Proponents of DTC advertising claim that it provides valuable product-related information to health care professionals and patients, may contribute to better use of medications, and helps patients take charge of their own health care. Opponents argue that DTC advertising provides misleading messages rather than well-balanced, evidence-based information. The literature is replete with opinions about the effects of prescription drug advertising on pharmaceutical drug prices and physician-prescribing patterns, but few studies have addressed the issues beyond opinion surveys. The economic literature on advertising effects in other markets, however, may provide insight. CONCLUSION: DTC advertising indirectly affects the price and the quantity of production of pharmaceuticals via its effect on changes in consumer demand. PMID- 12637130 TI - A structured clinical model for predicting the probability of pulmonary embolism. AB - PURPOSE: To develop a structured model to predict the clinical probability of pulmonary embolism. METHODS: We studied 1,100 consecutive patients with suspected pulmonary embolism in whom a definite diagnosis had been established. We used logistic regression analysis to estimate the probability of pulmonary embolism based on patients' clinical characteristics; the probability was categorized as low (< or = 10%), intermediate (>10%, < or = 50%), moderately high (>50%, < or = 90%), or high (>90%). RESULTS: The overall prevalence of pulmonary embolism was 40% (n = 440). Ten characteristics were associated with an increased risk of pulmonary embolism (male sex, older age, history of thrombophlebitis, sudden onset dyspnea, chest pain, hemoptysis, electrocardiographic signs of acute right ventricular overload, radiographic signs of oligemia, amputation of the hilar artery, and pulmonary consolidation suggestive of infarction), and five were associated with a decreased risk (prior cardiovascular or pulmonary disease, high fever, pulmonary consolidation other than infarction, and pulmonary edema on the chest radiograph). With this model, 432 patients (39%) were rated a low probability, of whom 19 (4%) had pulmonary embolism; 283 (26%) were rated an intermediate probability, of whom 62 (22%) had pulmonary embolism; 72 (7%) were rated a moderately high probability, of whom 53 (74%) had pulmonary embolism; and 313 (28%) were rated a high probability, of whom 306 (98%) had pulmonary embolism. CONCLUSION: This prediction model may be useful for estimating the probability of pulmonary embolism before obtaining definitive test results. PMID- 12637131 TI - Coagulation and activation of inflammatory pathways in the development of functional decline and mortality in the elderly. AB - PURPOSE: This study was performed to determine the effects of markers of inflammation (interleukin 6) and coagulation (D-dimer) on mortality and functional status in older persons. METHODS: Subjects were selected for the Duke Established Populations for Epidemiologic Studies of the Elderly. In 1992, 2569 subjects (age >71 years) were interviewed, of whom 1,723 had interleukin-6 and D dimer measurements. Values of interleukin 6 and D-dimer were categorized into quartiles. Outcomes were mortality (through 5 years) and functional status (through 4 years). Relative risks were estimated with proportional hazards models that adjusted for potential confounders. RESULTS: The relative risk of mortality was 1.28 (95% confidence interval [CI]: 0.98 to 1.69; P = 0.06) for those with only interleukin-6 levels in the highest quartile, 1.53 (95% CI: 1.18 to 1.97; P = 0.001) for subjects with only D-dimer levels in the highest quartile, and 2.00 (95% CI: 1.53 to 2.62; P = 0.0001) for those with levels of both in the highest quartile, as compared with those who were not in either of the highest quartiles. Those with high interleukin-6 and high D-dimer levels had the greatest declines in all measures of function. CONCLUSION: Activation of the coagulation and inflammatory pathways is associated with mortality and decline in function, and may be part of the explanation for the development of a frailty phenotype in the elderly. PMID- 12637132 TI - Correlation between ammonia levels and the severity of hepatic encephalopathy. AB - PURPOSE: Because the correlation between ammonia levels and the severity of hepatic encephalopathy remains controversial, we prospectively evaluated the correlation in 121 consecutive patients with cirrhosis. METHODS: The diagnosis of hepatic encephalopathy was based on clinical criteria, and the severity of hepatic encephalopathy was based on the West Haven Criteria for grading of mental status. Arterial and venous blood samples were obtained from each patient. Four types of ammonia measurements were analyzed: arterial and venous total ammonia, and arterial and venous partial pressure of ammonia. Spearman rank correlations (r(s)) were calculated. RESULTS: Of the 121 patients, 30 (25%) had grade 0 encephalopathy (no signs or symptoms), 27 (22%) had grade 1, 23 (19%) had grade 2, 28 (23%) had grade 3, and 13 (11%) had grade 4 (the most severe signs and symptoms). Each of the four measures of ammonia increased with the severity of hepatic encephalopathy: arterial total ammonia (r(s) = 0.61, P < or = 0.001), venous total ammonia (r(s) = 0.56, P < or = 0.001), arterial partial pressure of ammonia (r(s) = 0.55, P < or = 0.001), and venous partial pressure of ammonia (r(s) = 0.52, P < or = 0.001). CONCLUSION: Ammonia levels correlate with the severity of hepatic encephalopathy. Venous sampling is adequate for ammonia measurement. There appears to be no additional advantage of measuring the partial pressure of ammonia compared with total ammonia levels. PMID- 12637133 TI - Pharmacokinetic dosing of aminoglycosides: a controlled trial. AB - PURPOSE: To evaluate whether individualized pharmacokinetic dosing of aminoglycosides can reduce nephrotoxicity and improve the outcome of patients with gram-negative sepsis. METHODS: We conducted a prospective controlled trial at a tertiary care university hospital. Eighty-one patients with suspected or documented gram-negative infections were enrolled. All were treated with either gentamicin or amikacin, according to clinical judgement. Patients were allocated to one of two groups based on the last digit (odd/even) of their identification number. In the study group (pharmacokinetic dosing) of 43 patients, plasma aminoglycoside levels were determined 1 hour after initiation of drug infusion and 8 to 16 hours later to estimate the elimination half-life and volume of distribution, from which the subsequent dosage schedule was calculated. Target peak plasma levels were 20 microg/mL for gentamicin and 60 microg/mL for amikacin. Target trough levels were <1 microg/mL for both drugs. The control group (fixed once-daily dosing) consisted of 38 patients who were prescribed single daily doses of gentamicin or amikacin. The primary endpoints were renal toxicity (> or = 25% increase in serum creatinine level or a serum creatinine level > or = 1.4 mg/dL) and 28-day mortality. RESULTS: The two study groups were similar in age, sex, indications for therapy, Acute Physiology and Chronic Health Evaluation (APACHE) II score, and clinical assessment at baseline. Although the pharmacokinetic group received significantly greater doses of aminoglycosides than did the once-daily group, the incidence of nephrotoxicity was significantly lower in the pharmacokinetic group (5% [2/43] vs. 21% [8/38], P = 0.03). There was no statistically significant difference in 28-day mortality (27% [12/43] vs. 22% [8/38], P = 0.3). CONCLUSION: These results suggest that individualized pharmacokinetic dosing of aminoglycosides reduces the incidence of nephrotoxicity and allows the use of greater doses of aminoglycosides. PMID- 12637135 TI - Atrial fibrillation as a risk factor for stroke recurrence. AB - BACKGROUND: Although atrial fibrillation is a well-known risk factor for ischemic stroke, the extent to which it increases the risk of stroke recurrence, particularly in elderly patients, is less certain. METHODS: We performed a retrospective cohort study of 915 patients aged 50 to 94 years who were admitted with an ischemic stroke. The rates of recurrent strokes and recurrent severe strokes were estimated with the Kaplan-Meier method. The effects of atrial fibrillation on stroke risk were analyzed with proportional hazards models. RESULTS: Of the 829 patients who survived the initial hospitalization, 163 (20%) had a stroke during follow-up. Of the 203 patients with atrial fibrillation during index hospitalization who were not anticoagulated, 54 (27%) had recurrent strokes, compared with 18% (19/103) among those with atrial fibrillation who were anticoagulated and 17% (90/523) among those without atrial fibrillation. The age adjusted hazard ratio for recurrent stroke among those with atrial fibrillation who were not treated with anticoagulants was 2.1 (95% confidence interval [CI]: 1.4 to 2.9; P <0.001), whereas the hazard ratio for recurrent severe stroke was 2.4 (95% CI: 1.6 to 3.6; P <0.001). The increased risk was observed even in patients aged > or = 80 years and persisted during the follow-up for more than 5 years. CONCLUSION: Atrial fibrillation was an independent risk factor for stroke recurrence over a wide age range. PMID- 12637134 TI - The relation of C-reactive protein levels to total and cardiovascular mortality in older U.S. women. AB - PURPOSE: To determine whether serum C-reactive protein levels, a sensitive indicator of inflammation, are associated with the risk of cardiovascular mortality among older women. METHODS: We conducted a case-cohort study within the Study of Osteoporotic Fractures, a population-based study involving 9,704 women aged > or = 65 years from four U.S. centers. We randomly selected 400 women from the entire cohort plus an additional random sample of 92 women from the 1,125 women in the cohort who had died during the first 6 years of follow-up. Baseline serum C-reactive protein levels were measured using a high-sensitivity immunoassay. Cause-specific mortality was ascertained by review of death certificates and hospitalization records. Multivariable Cox proportional hazards regression was used to determine the association between C-reactive protein levels and cardiovascular mortality. RESULTS: During 6 years of follow-up, 150 of the 492 women died, including 52 who died of cardiovascular disease. After adjusting for potential confounders, women with C-reactive protein levels in the highest quartile (>3.0 mg/L) had a 8.0-fold (95% confidence interval [CI]: 2.2 to 29) greater risk of cardiovascular mortality than those in the lowest quartile (< or = 1.0 mg/L). The association remained strong in women who did not smoke or take estrogen, and when early deaths were excluded. Women who smoked and whose C reactive protein levels were above the first quartile had a very high risk of cardiovascular mortality (relative risk [RR] = 13; 95% CI: 3.4 to 47). C-reactive protein levels were not associated with noncardiovascular mortality (RR = 0.92; 95% CI: 0.4 to 2.1). CONCLUSION: C-reactive protein level was an independent predictor of cardiovascular mortality in older women. PMID- 12637136 TI - Eosinophilic meningitis. AB - Eosinophilic meningitis is a rare clinical entity that can be useful in narrowing the differential diagnosis of central nervous system disease. It is defined by the presence of 10 or more eosinophils/microL in the cerebrospinal fluid (CSF) or a CSF eosinophilia of at least 10%. The most common cause is invasion of the central nervous system by helminthic parasites, particularly Angiostrongylus cantonensis, but other infections as well as noninfectious conditions may also be associated. This review describes the etiologies of eosinophilic meningitis, focusing primarily on the helminths that cause CSF eosinophilia. PMID- 12637137 TI - Association between troponin I levels and mortality in stable hemodialysis patients. PMID- 12637138 TI - No association between the TGF-beta 1 Leu10Pro polymorphism and osteoporosis among white women in the United States. PMID- 12637139 TI - Before diagnostic testing for pulmonary embolism: estimating the prior probability of disease. PMID- 12637140 TI - Inflammation and coagulation: direct mediators of physical function in the elderly? PMID- 12637141 TI - Ammonia levels and the severity of hepatic encephalopathy. PMID- 12637142 TI - Once-daily dosing of aminoglycosides: how much monitoring is truly required? PMID- 12637143 TI - C-reactive protein and the risk of cardiovascular mortality. PMID- 12637144 TI - Clinical and pathological recognition of leprosy. PMID- 12637145 TI - Psoas abscess due to Mycobacterium xenopi infection. PMID- 12637146 TI - Evolving internal medicine faculty and resident comprehension of and attitudes toward a departmental documentation compliance program (1999-2002). PMID- 12637147 TI - NMR Nomenclature: Nuclear Spin Properties and Conventions for Chemical Shifts. IUPAC Recommendations 2001. AB - A unified scale is recommended for reporting the NMR chemical shifts of all nuclei relative to the (1)H resonance of tetramethylsilane. The unified scale is designed to provide a precise ratio, Xi, of the resonance frequency of a given nuclide to that of the primary reference, the (1)H resonance of tetramethylsilane (TMS) in dilute solution (volume fraction, varphi<1%) in chloroform. Referencing procedures are discussed, including matters of practical application of the unified scale. Special attention is paid to recommended reference samples, and values of Xi for secondary references on the unified scale are listed, many of which are the results of new measurements. Some earlier recommendations relating to the reporting of chemical shifts are endorsed. The chemical shift, delta, is redefined to avoid previous ambiguities but to leave practical usage unchanged. Relations between the unified scale and recently published recommendations for referencing in aqueous solutions (for specific use in biochemical work) are discussed, as well as the special effects of working in the solid state with magic-angle spinning. In all, nine new recommendations relating to chemical shifts are made. Standardised nuclear spin data are also presented in tabular form for the stable (and some unstable) isotopes of all elements with non-zero quantum numbers. The information given includes quantum numbers, isotopic abundances, magnetic moments, magnetogyric ratios, and receptivities, together with quadrupole moments and line-width factors (where appropriate). PMID- 12637148 TI - Genetic alterations in cancer as a result of breakage at fragile sites. AB - The organization and replication of DNA render fragile sites (FSs) prone to breakage, recombination as well as becoming preferential targets for mutagens carcinogens and integration of oncogenic viruses. For many years, attempts to link FSs and cancer generated mostly circumstantial evidence. The discoveries that chromosome translocations, amplification of proto-oncogenes, deletion of tumor suppressor genes, and integration of oncogenic viruses all result from the specific breakage of genomic DNA at FSs, however, have provided compelling support for such a link, further suggesting a causative role for FSs in cancer. PMID- 12637149 TI - Potentiation of anti-cancer effect by intravenous administration of vesiculated alpha-tocopheryl hemisuccinate on mouse melanoma in vivo. AB - We examined the effect of alpha-tocopheryl hemisuccinate (TS) on the growth of mouse melanoma cells B16-F1 inoculated on the back of hairless mice by two administration procedures of TS, i.p. administration of TS dissolved with dimethyl sulfoxide (TS i.p.) and i.v. administration of TS vesicles (TS-vesicle i.v.). TS i.p. significantly prevented the tumor growth of only half the mice in the group. However, TS-vesicle i.v. almost completely inhibited the tumor growth of all mice. Furthermore, the mean survival of the TS-vesicle i.v. group was 1.4 fold those of the control and TS i.p. groups. PMID- 12637150 TI - Enhancement of lung carcinogenesis by nonylphenol and genistein in a F344 rat multiorgan carcinogenesis model. AB - The modifying effects of nonylphenol and genistein on cancer induction were assessed in a multi-organ carcinogenesis model in male F344 rats initially treated with five different carcinogens. In experiment 1 rats received 250 or 25 ppm nonylphenol, or 250 or 25 ppm genistein in their diet for 28 weeks. The total incidences of adenomas and carcinomas in the lungs of animals treated with nonylphenol and genistein were significantly higher than in the control group. 5 Bromo-2'-deoxyuridine labeling indices, reflecting cell proliferation, were also significantly elevated in the lungs of rats given 250 and 25 ppm nonylphenol and 250 ppm genistein. In experiment 2, rats were treated with nonylphenol or genistein at concentrations of 250 ppm after DHPN initiation. In the lung, formation of 8-hydroxy-2'-deoxyguanosine, a marker of oxygen radical-mediated DNA damage, was significantly increased. These results indicate that nonylphenol and genistein have the potential to promote rat lung carcinogenesis, possibly via a mechanism involving stimulation of cell proliferation and DNA damage caused by oxygen radicals. PMID- 12637151 TI - Enhancing risk of ethanol on MeIQx-induced rat hepatocarcinogenesis is accompanied with increased levels of cellular proliferation and oxidative stress. AB - We investigated promotion potential of ethanol after initiation of hepatocarcinogenesis in male, 21-day-old, F344 rats by exposure to 10 ppm 2-amino 3, 8-dimethylimidazo[4,5-f]quinoxaline pellet diet for 8 weeks. The rats in group 1 were then fed on liquid control diet for 16 weeks, group 2 receiving the same diet containing 5% ethanol for 8 weeks followed by 8 weeks on the control diet, while group 3 animals were given 5% ethanol containing liquid diet for the entire16 weeks. On sacrifice at the end of week 24, glutathione S-transferase placental form positive foci, putative preneoplastic lesions in the liver, cell proliferation as indicated by proliferating cell nuclear antigen immunohistochemical staining and levels of 8-hydroxydeoxyguanosine, a marker of oxidative DNA damage, were significantly increased in the liver of group 3 along with non significant alteration of 8-oxoguanine DNA glycosylase mRNA expression. Lack of persistent increase of above parameters was found in transient ethanol exposure group. These results suggest that chronic consumption of ethanol promotes hepatocarcinogenesis by increasing oxidative stress and cell proliferation. It is also evident that abstinence of ethanol during the second stage stops its persistent promotion effect. PMID- 12637152 TI - Epstein-Barr virus and human hepatocellular carcinoma. AB - Recent studies suggest that Epstein-Barr virus (EBV) may act as a helper virus for the development of hepatocellular carcinoma by promoting replication of the hepatitis C virus (HCV) in the infected liver. Detection of EBV DNA in a high percentage of HCV-positive human hepatocellular carcinomas (HCC) from Japanese patients has supported this concept. In order to determine whether EBV infection is associated with HCC, we examined paraffin-embedded tissues from 31 cases of non-cirrhotic livers with hepatocellular carcinoma for the presence of EBV, HCV and hepatitis B virus (HBV) infection. RNA prepared from tumor samples were used as a template for reverse transcription followed by double-nested PCR with primers for the 5' untranslated region (NT) of HCV. DNA extracts of tumor samples were tested by single polymerase chain reaction for the detection of EBV and HBV (X- and/or S-gene) DNA sequences. To control for nucleic acid integrity, all tumor samples were amplified for human beta-globin DNA by polymerase chain reaction and subjected to Southern blot hybridization. None of the cases was found to be positive for EBV. Ten HCC cases (32%) tested positive for HCV and 12 HCC cases (38%) tested positive for HBV. Six of the surveyed patients had nucleic acids of both HCV and HBV in their tumor tissue. All HCC tumor samples were positive for beta-globin. Our study shows that HCV and HBV infections, but not EBV infection, are associated with hepatocarcinogenesis in non-cirrhotic livers. Other unknown risk factors seem to be in effect in the development of hepatocellular carcinoma in non-cirrhotic livers. PMID- 12637153 TI - Purification of normal lymphocytes from leukemic T-cells by lectin-affinity adsorbents - correlation with lectin-cell binding. AB - Utilization of leukemic T-cells from normal ones, using lectin-affinity adsorbents, is described. CNBr-activated Sepharose 6MB was covalently coupled to Soybean (SBA) or Dolichos Biflorus Agglutinins (DBA), then serves as an affinity probe for separation of leukemic T-cells from normal lymphocytes. The normal lymphocytes were removed almost completely by phosphate buffered saline (Ca(2+) and Mg(2+) free) (PBS(-)) from lectin-affinity column. More than 80% of the leukemic T-cells were retained on the lectin-affinity adsorbent, whereas another 10-15% were easily removed by PBS(-). There was a very good linear correlation between percent of cells, retained on the lectin-affinity adsorbent and percent of cells, interacting with the respective free lectin (r=0.97 for SBA, and r=0.93 for DBA). The viability of normal lymphocytes was not influenced after passing through the columns. In the case of leukemic T-cells - about 90% of the easily removed cells were dead, and another 10% were viable cells, non-interacting with DBA or SBA. PMID- 12637154 TI - A constitutively active ErbB4 mutant inhibits drug-resistant colony formation by the DU-145 and PC-3 human prostate tumor cell lines. AB - ErbB4 (HER4) is a member of the ErbB family of receptor tyrosine kinases, a family that also includes the Epidermal Growth Factor Receptor (EGFR/ErbB1/HER1), Neu/ErbB2/HER2, and ErbB3/HER3. Several groups have hypothesized that signal transduction by the ErbB4 receptor tyrosine kinase is coupled to differentiation, growth arrest, and tumor suppression in mammary and prostate epithelial cells. In this report we demonstrate that a constitutively active ErbB4 mutant inhibits the formation of drug-resistant colonies by the DU-145 and PC-3 human prostate tumor cell lines. This is consistent with our hypothesis that ErbB4 signaling is growth inhibitory and may be coupled to tumor suppression in prostate cells. PMID- 12637155 TI - Mutation of the DNA methyltransferase (DNMT) 1 gene in human colorectal cancers. AB - Alteration of DNA methylation is one of the most consistent epigenetic changes in human cancers. DNA methyltransferase (DNMT) 1 is a major enzyme that determines genomic methylation patterns. In order to understand the significance of mutations of the DNMT1 gene during human carcinogenesis, we performed polymerase chain reaction-single strand conformation polymorphism analysis using 46 oligonucleotide primer sets for all 40 coding exons and the 5'-flanking region (450 bp) of the DNMT1 gene in 29 colorectal cancers, 32 stomach cancers, 40 hepatocellular carcinomas (HCCs) and a corresponding sample of non-cancerous tissue from each case. Mutations in coding exons of the DNMT1 gene were detected in two (7%) of the colorectal cancers: they consisted of one-base deletion resulting in deletion of the whole catalytic domain and a point mutation resulting in a single amino acid substitution. No stomach cancers or HCCs showed mutations in the coding exons of the DNMT1 gene. No mutation in the 5'-flanking region of the DNMT1 gene was detected in any of the colorectal and stomach cancers or HCCs. These data suggest that mutational inactivation of the DNMT1 gene that potentially causes a genome-wide alteration of DNA methylation status may be a rare event during human carcinogenesis. PMID- 12637156 TI - p130 expression in thyroid neoplasms: its linkage with tumor size and dedifferentiation. AB - p130 belongs to the retinoblastoma-related gene family, and its gene product works to negatively regulate cell cycle progression in the G1 phase. In this study, we investigated p130 expression in thyroid neoplasms. p130 overexpression was observed in 33.3% of follicular adenoma and 50% of follicular carcinoma and the incidences were not significantly different. In papillary carcinoma, it was overexpressed in 36.5% of cases, but in microcarcinoma, the incidence was significantly lower (14.3%). Furthermore, in anapalstic carcinoma, p130 overexpression was less frequently seen than in papillary carcinoma except for microcarcinoma and follicular carcinoma, and only 14.3% of cases overexpressed this protein. These findings suggest that: (1) reduced p130 expression may contribute to the aggressive character of anplastic carcinoma; and (2) p130 may specify the growth characteristics of microcarcinoma. PMID- 12637157 TI - Immunohistochemical expression of C-reactive protein in squamous cell carcinoma of the esophagus - significance as a tumor marker. AB - C-reactive protein (CRP) is a prototype acute phase protein which has been known to be synthesized in hepatocytes. Although serum elevation of CRP has been reported to be an indicator of the unfavorable outcome of the patients in some malignant tumors, the expression of the protein in carcinoma cells has not been investigated. The aim of the current study was to assess the immunohistochemical expression of CRP in squamous cell carcinoma (SCC) of the esophagus and to find its biological significance. Immunohistochemical examination for CRP expression was performed for 37 advanced esophageal SCCs with the depth of T2, T3 or T4, which had been surgically resected without preoperative therapy for the patients. Eighteen carcinomas (48.6%) demonstrated immunohistochemical CRP expression. Univariate analysis showed that the prognosis of the patients with esophageal SCCs expressing CRP was significantly worse than that in patients with tumors without CRP expression (P=0.017). Moreover, CRP expression was found to be an independent prognosticator in patients with esophageal SCCs in the multivariate analysis (P=0.036). To the best of authors' knowledge, this is the first report that demonstrated the possible carcinoma-related expression of CRP in SCCs of the esophagus and its biological significance as the prognosticator of the patients. PMID- 12637158 TI - Quantitative assessment of glioblastoma invasion in vivo. AB - The aim of this study was the quantitative assessment of the time course and spatial distribution in brain of invading glioblastoma (GBM) cells using a recently described model consisting of RT2 rat GBM cells stably transfected with enhanced green fluorescent protein (eGFP) - called RT2-eGFP - and implanted in Fischer rats. Invasion throughout the brain was verified by confocal microscopy and immunocytochemical staining for eGFP. Rats were sacrificed on post implantation days 3, 8, and time of death (TOD). First, the entire rat brain was disaggregated at each time point and viable RT2-eGFP cells were counted using flow cytometry with fluorescence as the marker. Next, 2 mm(3) samples of cortex from each of four brain quadrants (bifrontal and bioccipital) were disaggregated at each time point, with tumor cell quantification as before. Tumor cell density, averaged over the entire brain, reached a peak mid-way through its time course, leveling out by TOD. Tumor cell density within bulk tumor (BT) was greatest early in the evolution of the brain tumor, decreasing to its final value mid-way through its time course, due to necrosis. The greatest concentration of tumor cells was within BT, with up to an order of magnitude fewer cells in the periphery, while the number of brain tumor cells invading brain distant from BT remained constant from day 3 until TOD. BT size steadily increased after implantation, with an increasing portion due to central necrosis as time progressed, suggesting that this effect is an important contributor to fatality in this model. Alternatively (or additionally), accumulation of toxins elaborated by tumor cells throughout the brain starting early in the evolution of the tumor may also contribute to fatality. PMID- 12637160 TI - Swelling of intercellular lipid lamellar structure with short repeat distance in hairless mouse stratum corneum as studied by X-ray diffraction. AB - Lamellar structures of intercellular lipids in stratum corneum of hairless mouse were studied at various water contents by small-angle X-ray diffraction. At room temperature there are at least two lamellar structures, long and short lamellar structures, with repeat distances of 13.6 and around 6 nm, respectively. The long lamellar spacing is almost constant over the water content from 0% w/w to 80% w/w that is consistent with the previously reported results. For the short lamellar structure we found that with increasing the water content the lamellar spacing becomes larger, that is, from 12 to 50% w/w the short lamellar spacing increases from 5.8 to 6.6 nm. In addition to the previously reported result that at the water content of about 20% w/w the X-ray diffraction peak for the long lamellar structure becomes sharp, we found that this is also the case for the short lamellar structure. Below the water content of about 12% w/w the X-ray diffraction peak for the short lamellar structure dies out and conversely above the water content of about 50% w/w it becomes weak and finally merges into the second-order diffraction peak for the long lamellar structure. Considering the matching of the long lamellar spacing that is unchanged with the water content and twice the short lamellar spacing that changes as a function of the water content, it is likely that the swelling of the short lamellar structure plays an important role in the regulation of water stored in stratum corneum. PMID- 12637159 TI - ING1 and p53 tumor suppressor gene alterations in adenocarcinomas of the esophagogastric junction. AB - The aim of this study was to characterize molecular alterations of the recently reported candidate tumor suppressor gene, ING1, and to explore the relationship between ING1 and p53 in a well-defined series of adenocarcinomas of the esophagogastric junction (AdEGJ). Polymerase chain reaction (PCR)-based assays were used to characterize ING1 and p53 alterations, relative to histologically normal esophageal mucosa. Two tumors were found to have ING1 mutations: one novel missense mutation (AGC(Ser)-->ATC(Ile)) at codon 147, and one silent mutation (TCG(Ser)-->TCA(Ser)) at codon 173. Reduced expression of the two major alternatively spliced ING1 messenger RNA variants, p47(ING1a) and p33(ING1b) was variable, but was reduced (1.2-10-fold) in 12 of 19 AdEGJs compared to normal esophageal epithelium. No association between p53 and ING1 alterations was apparent. We conclude that reduced ING1 expression is frequently associated with AdEGJ tumorigenesis, further supporting its role as a tumor suppressor gene, and that ING1 expression is independent of p53 status. PMID- 12637161 TI - Polymorphism of ceramide 3. Part 1: an investigation focused on the head group of N-octadecanoylphytosphingosine. AB - The thermotropic phase behaviour of the ceramide N-octadecanoylphytosphingosine (CER3) was investigated using differential scanning calorimetry, X-ray powder diffraction and FT-IR spectroscopy. CER3 was shown to be a polymorphic substance depending on the crystallisation conditions. Three different solid states were found. The FT-IR results elucidate changes in the hydrogen bonding interactions of the ceramide head group. It was shown that the amide I and the amide II vibration bands are quite sensitive to the phase transitions of CER. There are clear shifts in the band positions of those bands passing the phase transitions. Furthermore, changes were observed in the NH- and OH- stretching region. The study shows that there are strong inter- and intramolecular hydrogen bonds between hydroxy groups in the ceramide head group. There are also strong hydrogen bonds to the amide oxygen as shown by the band positions of the amide vibrations. The H-bonding network and conformation of the head group of CER3 alters due to the phase transitions. PMID- 12637162 TI - Phase behavior in multilamellar vesicles of DPPC containing ganglioside GM3 with a C18:1 sphingoid base and a 24:0 acyl chain (GM3(18,24)) observed by X-ray diffraction. AB - Structures and phase behavior of multilamellar vesicles of 1,2-dipalmitoyl-L phosphatidylcholine (DPPC) containing various amount of ganglioside GM3 with a C18:1 sphingoid base and a 24:0 acyl chain (GM3(18,24)) were investigated by small-angle X-ray diffraction. Below 3.5 mol% GM3 content, the phase behavior was similar to that of pure DPPC except for a slight increase of lamellar repeat distance in the L(beta'), the P(beta') and the L(alpha) phases and a decrease of the pretransition temperature. In the range of 4-12 mol% GM3 content, another phase which has larger repeat distances coexisted with the phase observed below 3.5 mol% GM3 content. This has been interpreted that the phase separation into GM3-poor phase (denoted as A-phase) and GM3-rich phase (denoted as B-phase) took place. Above 13 mol% GM3 content, the B-phase became dominant. This phase separation may be related to the formation of GM3-enriched microdomains that had been observed on the cell surfaces which express large amounts of GM3, such as murine B16 melanoma (J. Biol. Chem. 260 (1985) 13328). PMID- 12637163 TI - Does sucrose influence the properties of DMPC vesicles? AB - Small-angle neutron and X-ray scattering, dynamic light scattering, X-ray diffraction coupled with differential scanning calorimetry, and Raman spectroscopy were applied to investigate unilamellar (ULVs) and multilamellar (MLVs) dimyristoylphosphatidylcholine (DMPC) vesicles in aqueous sucrose solutions with sucrose concentrations from 0 to 60% w/w. In case of ULVs, the addition of sucrose decreases the polydispersity of vesicle population. A minimum value of polydispersity was found at 20% sucrose. For sucrose concentration from 0 to 35% oligolamellar vesicles in the ULV population have a minimum presence. Vesicles with 5-10% sucrose exhibit the best stability in time. For the case of MLVs, sucrose influences the temperature of the phase transitions, but the internal membrane structure remains unchanged. PMID- 12637164 TI - Observation of complex thermal transitions for mixed micelle solutions containing alkyldimethylphosphine oxides and phospholipids and the accompanying cloud points. AB - The thermal properties of various mixtures of two nonionic surfactants, decyldimethylphosphine oxide (APO10) and dodecyldimethylphosphine oxide (APO12) and two phospholipids, dimyristoylphosphatidyl choline (DMPC) and dipalmitoylphosphatidyl choline (DPPC), were examined by differential scanning calorimetry at various mole fractions. The addition of APO12 to DMPC multilamellar vesicles lowered the temperature of the main transition, produced considerable broadening, and eliminated the pre-transition. Phase separation, as evidenced by the existence of a cloud point, T(cp), occurred when the mole fraction of APO12, with respect to DMPC was 0.58 and above. A small abrupt increase in heat capacity was observed at, or slightly above, the cloud point of APO12 and all mixed micelle solutions. It appeared that mixed micelles coexisted with mixed bilayers when the mole fraction was between 0.58 and 0.75 and perhaps as low as a mole ratio of 0.32. All of the mixtures, except APO12/DMPC, exhibited a clear endotherm below the temperature corresponding to the cloud point, which likely reflects the growth in micellar size. Overlapping chain length dependent endothermic peaks, perhaps resulting from reorganization and/or continued association of the micelles, were observed above the cloud point for all of the mixtures except for APO10/DMPC solutions. However, solutions of mixed micelles consisting of APO10/DMPC with mole fractions of surfactant between 0.81 and 0.93 portrayed a broad unidentified exotherm of about 2+/-1 kcal/mol, which was centered nearly 10-20 degrees C above the cloud point. PMID- 12637165 TI - Comparative study on the action of tocopherols and tocotrienols as antioxidant: chemical and physical effects. AB - alpha-Tocopherol is known as the most abundant and active form of vitamin E homologues in vivo, but recently the role of other forms of vitamin E has received renewed attention. The antioxidant properties were compared for alpha-, beta-, gamma- and delta-tocopherols and tocotrienols. The following results were obtained: (1). the corresponding tocopherols and tocotrienols exerted the same reactivities toward radicals and the same antioxidant activities against lipid peroxidation in solution and liposomal membranes; (2). tocopherols gave more significant physical effect than tocotrienols on the increase in rigidity at the membrane interior; (3). tocopherols and tocotrienols showed similar mobilities within the membranes, but tocotrienols were more readily transferred between the membranes and incorporated into the membranes than tocopherols; (4). alpha tocopherol and alpha-tocotrienol, but not the other forms, reduced Cu(II) to give Cu(I) together with alpha-tocopheryl and alpha-tocotrienyl quinones, respectively and exerted prooxidant effect in the oxidation of methyl linoleate in SDS micelles. PMID- 12637166 TI - Peroxyl radicals promoted changes in water permeability through gramicidin channels in DPPC and lecithin-PC vesicles. AB - Gramicidin incorporation to DPPC or lecithin-PC large unilamellar vesicles (LUVs) leads to pore formation that, under hyper-osmotic conditions, produces a noticeable increase in the rate of trans-membrane water flow. This pore formation is more efficient in the more fluid lecithin-PC LUVs. Exposure of these vesicles to peroxyl radicals generated in the aerobic thermolysis of 2,2'-azo-bis(2 amidinopropane) (AAPH), changes the physical properties of the bilayer (as sensed employing fluorescent probes), modifies gramicidin molecules (as sensed by the decrease in Trp fluorescence) and notably reduces the transbilayer rate of water outflow. In order to evaluate if this reduced water-transport capacity is due to changes in the membrane due to lipid-peroxidation and/or direct damage to gramicidin channels, results obtained in the oxidable vesicles (lecithin-PC) were compared to those obtained in DPPC vesicles. The data obtained show that most of the water transport efficiency loss can be ascribed to a direct disruption of gramicidin channels by AAPH derived peroxyl radicals. PMID- 12637167 TI - Towards redox active liquid crystalline phases of lipids: a monoolein/water system with entrapped derivatives of ferrocene. AB - The phase and electrochemical behavior of the aqueous mixtures of monoolein (MO) and synthetic ferrocene (Fc) derivatives containing long alkyl chains-(Z)-octadec 9-enoylferrocene (1), (Z)-octadecen-9-ylferrocene (2), and ferrocenylmethyl (Z) octadec-9-enoate (3)-were studied. At low hydration, the reversed micelles (L(2) phase) and cubic Q(230) phase of MO can accommodate relatively high amounts (>6 wt.%) of the Fc-derivative 2, whereas at high hydration, the pseudoternary cubic phase Q(224) is destabilized even at about 2 wt.% of this Fc. Increasing the Fc derivative content induces L(alpha)-->L(2) and L(alpha)-->reversed bicontinuous cubic phase (Q(II))-->H(II) transitions depending upon hydration. A rough study of the MO system containing compounds 1 and 3 indicates very similar phase behavior to that of the MO/2/H(2)O system. Compound 2 apparently has no effect on the lipid monolayer thickness in the pseudoternary L(alpha), H(II) and Q(II) liquid crystalline phases of MO. Within a 3D-structure of the Q(224) phase, derivatives 1-3 exhibit electrochemical activity on the gold electrode. The one electron redox conversion processes are electrochemically quasi-reversible and controlled by diffusion. The values of apparent diffusion coefficient (D(app)) and heterogeneous electron-transfer rate constant (k(s)) of Fcs are significantly lower in the cubic phase matrix when compared to the acetonitrile solution. By contrast, the MO H(II) phase with entrapped Fc-derivatives does not exhibit electrochemical activity on the electrode surface. It is suggested that the diffusional anisotropy and/or localized aggregation of compounds 1-3 within a 2D structure of the H(II) phase account(s) for the latter observation. PMID- 12637168 TI - A more detailed fatty acid composition of human lipoprotein(a)--a comparison with low density lipoprotein. AB - Lipoprotein(a)'s (Lp(a)'s) fatty acid composition is partially known for the cholesteryl ester (CE), triglyceride (TG) and total phospholipid (PL) fractions. Individual PLs' fatty acids are unknown. This study sought to confirm and extend existing data and elucidate the individual PLs of Lp(a). For Lp(a) versus LDL, the mole percentage saturated fatty acids comprised 11.3+/-1.3 versus 16.8+/-1.2 (CE) (P<0.05), 43.4+/-5.2 versus 39.2+/-4.0 (TG) (P<0.05), 55.7+/-6.3 versus 54.7+/-5.9 (PL) (P>0.05), 51.9+/-3.5 versus 50.2+/-4.2 (choline-containing phospholipids (PC)) (P>0.05), 40.2+/-4.6 versus 43.1+/-3.9 (ethanolamine containing phospholipids (PE)) (P>0.05), 73.2+/-7.6 versus 81.2+/-8.2 (sphingomyelin (SPH)) (P<0.05). Linoleic acid was CE's major fatty acid and while palmitic acid was the major fatty acid in all other fractions except PE. PMID- 12637169 TI - Solvent accessible surface area (ASA) of simulated phospholipid membranes. AB - The membrane-solvent interface has been investigated through calculations of the solvent accessible surface area (ASA) for simulated membranes of DPPC and POPE. For DPPC at 52 degrees C we found an ASA of 126+/-8 A(2) per lipid molecule, equivalent to twice the projected lateral area. The ASA was dominated by the contribution from the quaternary ammonium ion which accounts for about two thirds of the total value. The remaining ASA is primarily the phosphate moiety (22+/-1 A(2)), and this contribution almost entirely represents the two 'out-of-chain' oxygen atoms. Interestingly, even the most exposed parts of the PC head-group show average ASAs of less than half of its maximal or 'fully hydrated' value. The average ASA of a simulated POPE membrane was 96+/-7 A(2) per lipid. The smaller value than for DPPC reflects much lower ASA of the ammonium ion, which is partially compensated by increased exposure of the ethylene and phosphate moieties. The ASA of the polar moieties of (PO(4), NH(3) and COO) constitutes 65% of the total accessible area for POPE, making this interface more polar than that of DPPC. It is suggested that ASA information can be valuable in attempts to rationalize experimental investigations, particularly molecular interpretations of thermodynamic information. PMID- 12637171 TI - The physiological basis of clinical deficits in Parkinson's disease. AB - Despite the fact that Parkinson's disease (PD) is a relatively common neurological condition, the physiological derangements that result in its clinical features remain unclear. On combining findings from psychophysical, clinical and electrophysiological studies, an overriding theme is proposed that PD deficits are essentially quantitative rather than qualitative in nature. This may arise because the normal function of the basal ganglia is to activate neural processes selectively, providing appropriate diversion of "attentional" resources for decision-making aspects of motor tasks and appropriate "energising" of the executive aspects of such tasks. It is suggested that these concepts of attention, an idea stemming from psychophysical studies, and of energisation, which has derived from kinematic studies, may in fact reflect the same universal process of selective facilitation of particular processes and inhibition of others. In PD, without efficient facilitation, tasks may still be performed but less well than in normal individuals. Possible underlying mechanisms of basal ganglial function are discussed in the context of new findings on direct and indirect pathway actions and the role that oscillatory modulations may play in achieving selective facilitation is explored. Further investigation of disturbances of such mechanisms in PD may prove important in understanding the underlying pathophysiology of the condition. PMID- 12637170 TI - Neural mechanisms and behaviors for acoustic communication in teleost fish. AB - Sound communication is not unique to humans but rather is a trait shared with most non-mammalian vertebrates. A practical way to address questions of vocal signal encoding has been to identify mechanisms in non-mammalian model systems that use acoustic communication signals in their social behavior. Teleost fishes, the largest group of living vertebrates, include both vocal and non-vocal species that exploit a wide range of acoustic niches. Here, we focus on those vocal species where combined behavioral and neurobiological studies have recently begun to elucidate a suite of adaptations for both the production and the perception of acoustic signals essential to their reproductive success and survival. Studies of these model systems show that teleost fish have the vocal-acoustic behaviors and neural systems both necessary and sufficient to solve acoustic problems common to all vertebrates. In particular, behavioral studies demonstrate that temporal features within a call, including pulse duration, rate and number, can all be important to a call's communicative value. Neurobiological studies have begun to show how these features are produced by a vocal motor system extending from forebrain to hindbrain levels and are encoded by peripheral and central auditory neurons. The abundance and variety of vocal fish present unique opportunities for parallel investigations of neural encoding, perception, and communication across a diversity of natural, acoustic habitats. As such, investigations in teleosts contribute to our delineating the evolution of the vocal and auditory systems of both non-mammalian and mammalian species, including humans. PMID- 12637173 TI - Effects of label format on knowledge acquisition and perceived readability by younger and older adults. AB - This research examines consumers' information acquisition and preference for labels of a simulated over-the-counter (OTC) medication. Twelve otherwise identical OTC drug bottles were compared with different back labels varying in (a) print size, (b) amount of white space between text, and (c) label design (standard vs extended/pull-out). A no back label condition served as a control. Older (mean age=77.7 years) and younger (mean age=21 years) adults were given one of the 12 bottles and asked to perform one of two information acquisition tasks: (a) they examined the bottle for 3 minutes and then completed a questionnaire with the bottle absent, or (b) they answered the same questionnaire while the bottle was present. Afterwards, participants were given all of the bottles and asked to rank them according to perceived readability. The younger adults' information acquisition performance was significantly better than the older adults' for all label conditions except the control condition where both groups' low performance did not differ. Specifically, the older adults' performance was significantly better in the medium and large print conditions than in the small print conditions--with the latter conditions not differing from the control condition. Younger adults showed no performance differences among the different print-size conditions. No substantial effects on knowledge acquisition performance from the white space manipulations were found. However, the perceived readability ranks showed that both groups preferred larger print size and white space. The white space effect was smaller than for print size, particularly for older adults. The extended/pull-out label design was facilitative for older adults in that it allowed the use of larger print. The results suggest that older consumers may be unable to acquire information in the 'fine' print frequently found in various kinds of product literature. PMID- 12637172 TI - Roles of the mammalian subventricular zone in brain development. AB - There has been enormous progress in uncovering the contributions of the subventricular zone (SVZ) to the developing brain. Here, we review the roles of four anatomically defined embryologic divisions of the SVZ of the mammalian brain: the lateral ganglionic eminence (LGE), the medial ganglionic eminence (MGE), the caudal ganglionic eminence (CGE), and the fetal neocortical SVZ (SVZn), as well as the roles of the two major anatomically defined regions of the postnatal SVZ, the anterior SVZ (SVZa) and the dorsolateral SVZ (SVZdl). We describe the types of cells within each subdivision of the SVZ, the types of brain cells that they generate during embryonic, fetal, and perinatal development, and when known the mechanisms that regulate their differentiation. This review provides a critical analysis of the literature, from which current and future studies on the SVZ can be formulated and evaluated. PMID- 12637174 TI - Pedestrian conspicuity research: a review. AB - This review sets out to evaluate the ways in which pedestrian conspicuity has been defined and measured and to consider the various scenarios in which studies of pedestrian conspicuity have been conducted. Research from the psychological and human factors literature is reviewed, in addition to relevant studies on conspicuity that fall outside the scope of applied psychology. Methodological differences between these studies are compared and their ecological validity in terms of the real-world context of pedestrians at risk from vehicles is discussed. The authors argue that there have been many methodological differences in pedestrian conspicuity studies, but that this may not necessarily be problematic when investigating a phenomenon with multiple causal factors. However, suggestions are made for improving ecological validity and establishing a more unified framework for future research in this area. PMID- 12637175 TI - A critical appraisal of current knowledge and future directions of ergophthalmology: consensus document of the ICOH Committee on 'Work and Vision'. AB - In the past, occupational health gave scant attention to the visual apparatus. Studies on issues relating to 'work and vision' have predominantly addressed industrial accidents and the toxic effects of exposure to chemical and physical agents, with secondary prevention being more common than primary prevention. In recent years, the huge transformations implemented in workplaces, mainly due to computer-based technologies, demand a progressively higher efficiency of the eye and the related nervous system components that co-ordinate eye movements and accommodation. At the moment these aspects are managed by medical, psychosocial and a variety of technical disciplines, without any shared criteria, terminology and method. Objectives of the Committee are: to investigate holistically the many possible hazards associated to the occupational visual performance; to come to a consensus on terminology, risk assessment and health surveillance procedures. The present status of research and practice in ergophthalmology does not allow for clear-cut decision about health risk and nor is there a clear direction on the prevention of work-related ocular and visual disturbances and disorders. Specifically, at least three determinants must be objectively considered, namely: (a) visual effort; (b) environmental agents; (c) individual characteristics. Although criteria exist for the evaluation of some of these determinants, a greater refinement is necessary 'for a good working life' of the visual apparatus. The ICOH Scientific Committee on 'Work and Vision' is seeking to systematically develop this matter, yielding a more rational and complete approach to the relationship between work and vision. PMID- 12637176 TI - Foot landmarking for footwear customization. AB - As consumers are becoming increasingly selective of what they wear on their feet, manufacturers are experiencing problems developing and fitting the right footwear. Literature suggests that shoes with a shape similar to feet may be comfortable because they attempt to maintain the feet in a neutral posture. The objective of this paper is to develop a metric to quantify mismatches between feet and lasts and also to be able to generate the two-dimensional outline of the foot using the minimum number of landmarks. Fifty Hong Kong Chinese were participants in the experiment. In addition to subject weight, height, foot length and foot width, the left foot outlines were drawn and 18 landmarks were marked on each of the two-dimensional foot outlines. A step-wise procedure was used to reduce the chosen 18 landmarks to eight, such that the mean absolute negative error (an indicator of 'tightness') between the foot outline and the modelled curve was 1.3 mm. These eight landmarks seem to show an improvement over those proposed by other researchers, thus showing the importance of choosing the right landmarks for modelling the foot. The positive and negative absolute errors were on average 1.8 mm and 1.3 mm respectively. Moreover, the mean errors for the toe region and for the rest of the foot were 1.7 mm and 1.6 mm respectively. The results indicate that the foot outline, an important component for footwear functionality and fitting, may be modelled using eight critical landmarks. PMID- 12637177 TI - Modular data logger system for physical workload measurements. AB - Direct technical measurements of physical workload have advantages in studies of work-related musculoskeletal disorders. A data logger has been developed, based on 20 and 40 Mbyte PCMCIA (Personal Computer Memory Card International Association) flash memory cards, for electromyography (EMG), inclinometry and goniometry. They are provided with an input for marking of events that can be used for synchronization with other loggers and with video recordings. To make the data easier to access by the preceeding analysis software, application software has also been developed that reorganizes the data. The loggers are easy to operate and enable whole-day ambulatory field recordings without any need for reduction or compression of the data. PMID- 12637180 TI - Development of a longitudinal study of complications and functional outcomes after traumatic brain injury. AB - PRIMARY OBJECTIVE: To create a longitudinal database of patients with moderate and severe traumatic brain injury. RESEARCH DESIGN: A prospective study design was used to collect data pertaining to demographics, acute and post-acute management, complications, resource utilization and functional outcomes. METHODS AND PROCEDURES: Data were collected on 233 patients with a Glasgow Coma Score of 12 or less, admitted to a Level 1 Trauma Centre within 24 hours of injury and continued through post-hospitalization follow-up. MAIN OUTCOMES AND RESULTS: The mean age was 37.7 years, 70% were males, 54% were motor vehicle related accidents, and 21% died. Of the 185 survivors, 23% were discharged directly home from acute hospital care and 74% required inpatient rehabilitation. At hospital discharge, 76% had Rancho Los Amigos Scores of VII or higher; 81% had no or only mild verbal communication deficits and 79% were able to ambulate. CONCLUSIONS: The study indicates that while it is difficult to predict functional outcomes for individual survivors of TBI in the early stages of acute care, they are often better than suspected at the time of injury. PMID- 12637181 TI - Epidemiology of traumatic comas: a prospective population-based study. AB - OBJECTIVE: Most studies on patients with severe brain injury (SBI) are based on data from specialized centres. This prospective epidemiologic study included all patients in a defined region with a coma lasting more than 24 hours or leading to a death. METHODS: All patients with a SBI admitted to an emergency department in the region were included during a 1-year period. A data form was completed with initial neurological state, CT scan lesions and associated injuries. Outcome at the end of acute hospitalization was assessed from medical notes. RESULTS: Two hundred and forty-eight patients were registered. Annual incidence was 8.5/100 000 population. Median age was 41 years. Traffic crashes were the most frequent cause (59%). Falls occurred in 30% (16% from a high level, 14% from one level). Initial GCS was above 8 in 31%, and patients with a neurological deterioration were older (52 vs 32 years). Death occurred in 52% of the cohort. Outcome was related to CT scan diagnosis, delay before eye opening and delay before obeying commands. CONCLUSION. This population-based cohort of patients with SBI was different from patients selected in trauma centres. The patients were older, more often injured in falls and their mortality rate remained very high. PMID- 12637182 TI - Evaluation of malingering cut-off scores for the Rey 15-Item Test: a brain injury case study series. AB - PRIMARY OBJECTIVE: To evaluate the utility of prescribed malingering cut-off scores by examining the Rey Fifteen-Item Test (FIT) responses of persons with severe brain injury. RESEARCH DESIGN: Series of five case studies. METHODS AND PROCEDURES: The FIT was administered along with standardized neuropsychological and neurobehavioural measures. MAIN OUTCOMES AND RESULTS: Using the standardized scoring protocol, all five patients obtained perfect (15/15 correct) scores on the FIT. All patients exhibited significant impairment on other neuropsychological indices, included measures of memory and attention. CONCLUSIONS: Results lend support to the use of higher cut-off scores to identify malingering. Further large-scale research is recommended to ascertain the reliability of FIT scoring procedures, determine cut-off scores for various diagnostic groups and develope age and education adjustments. Implications for use of the FIT in clinical and forensic settings are also discussed. PMID- 12637183 TI - Evaluation of extensibility, passive torque and stretch reflex responses in triceps surae muscles following serial casting to correct spastic equinovarus deformity. AB - PRIMARY OBJECTIVE: Spastic equinovarus deformity of the ankle in adults with acquired brain injury can severely limit the achievement of rehabilitation goals. This study examined changes in triceps surae muscle extensibility, passive resistive torque and soleus stretch reflex responses in 10 adult brain injured subjects undergoing serial casting to correct ankle equinovarus deformity. METHOD: Goniometric measurement of maximal passive dorsiflexion was used to evaluate extensibility of the triceps surae muscles. Computer controlled ankle dynamometry and surface electromyography were used to identify passive resistive torque and soleus stretch reflex onset angle in response to stretches at two velocities. RESULTS: The mean casting period was 5 weeks. Casting was discontinued in one subject due to failure to achieve measurable gain in ankle range over three consecutive cast changes. Median improvements in maximal ankle dorsiflexion, with the knee flexed or extended, of 30 degrees and 15 degrees, respectively, were achieved in the remaining nine subjects (p < 0.0001). The median passive ankle range in response to a displacing torque of 10 Nm increased 4.3 degrees over the intervention period (p < 0.0001). Consistent soleus reflex activity in response to passive stretches at 25 degrees x s(-1) was elicited in only four subjects. A trend for the stretch reflex onset to move further into the available range was demonstrated in these subjects. CONCLUSION: In the present study, serial casting contributed to significant change in triceps surae extensibility and passive resistive torque, corresponding with improved maximal passive ankle dorsiflexion range and an increase in the angle achieved with a displacing torque of 10 N.m. Increased stretch reflex threshold was observed in some subjects. The use of pre-determined outcome criteria and careful measurement of responses to this intervention were important to prevent premature discontinuation of casting when gains were slower than expected. PMID- 12637184 TI - Reliability of the self-awareness of deficits interview for adults with traumatic brain injury. AB - PRIMARY OBJECTIVE: This paper investigates the test-re-test reliability of the Self-Awareness of Deficits Interview (SADI) for assessing level of self-awareness in clients with traumatic brain injury (TBI). RESEARCH DESIGN: Twenty rehabilitation clients with TBI were interviewed using the SADI and re-test interviews were conducted 2-4 weeks later. METHODS AND PROCEDURES: Two checklists were used to collect collateral information on the client from significant others and rehabilitation staff. This information was used to assist in assigning SADI scores. MAIN OUTCOMES AND RESULTS: High test-re-test reliability was demonstrated for both total (ICC = 0.94) and sub-section scores (ICC = 0.85, 0.86 and 0.86). The results, combined with previous research indicating the high inter-rater reliability of the SADI, suggest the SADI is a reliable means of evaluating level of self-awareness. Further research is required to investigate other psychometric properties of the SADI and the two checklists. PMID- 12637185 TI - Agraphia with abnormal writing stroke sequences due to cerebral infarction. AB - A 65-year-old, right-handed man presented with speech and gait disturbances. He was alert and cooperative, showing mild right hemiparesis and sensory disturbance. Spontaneous speech was fluent; object naming, word fluency and reading were fully preserved. Sentence repetition and verbal comprehension were mildly impaired. Writing was slow, hesitant and difficult for both spontaneous writing and dictation. Copying was better, although he had some difficulty in copying letters and also complex figures. Sequences of strokes in forming written characters were abnormal; strokes were formed by piecing together several fragments. Computed tomography and magnetic resonance imaging showed a subcortical infarct in the left frontoparietal region. Characteristics of agraphia resembled 'apractic agraphia' and agraphia may have resulted largely from loss or unavailability of the memory of motor patterns necessary for writing letters. PMID- 12637186 TI - Treatment of oro-facial hypersensitivity following brain injury. AB - A 56 year old man who was 10 months post-severe traumatic brain injury was unable to tolerate oral hygiene. He had oro-facial hypersensitivity, oral dyspraxia and limited oral function. Poor oral hygiene with coating of oral structures and infection was present. An intensive systematic desensitization programme over 2 weeks, even at this late stage post-injury, increased oral tolerance and allowed full oral hygiene. Participation in oral hygiene and functional patterns of movement also improved, enabling some oral nutritional intake. This case study provides controlled evidence, very little of which exists in the literature, to demonstrate the effectiveness of these treatment techniques PMID- 12637187 TI - Rapid cycling bipolar disorder after left temporal polar damage. AB - The case of a 48-year-old woman with rapid cycling bipolar disorder subsequent to a traumatic brain injury is reported. Both depressive and manic episodes had an average duration of approximately 1 month, alternating without stable euthymic periods. Neuroradiological examinations revealed a circumscribed lesion in the left temporal pole. After 1 year without treatment, psychiatric intervention and pharmacotherapy was initiated. Her mood swings were successfully treated with the co-administration of valproate and lithium. Case reports of rapid cycling bipolar disorder after traumatic brain injury are very rare and this case supports the idea that temporal polar dysfunction is a candidate for the neurobiological basis of rapid cycling bipolar disorder. PMID- 12637188 TI - Geographic contiguity and genetic affinity among five ethnic populations of Manipur, India: further molecular studies based on VNTR and STR loci. AB - BACKGROUND: Apart from traditional markers studied among a few numerically small, geographically defined surveys among Mongoloid populations in northeastern parts of India, very little is known about their genomic diversity at the molecular level. PRIMARY OBJECTIVE: This study seeks to investigate how best the variable number tandem repeat (VNTR) and short tandem repeat (STR) loci together can detect the patterns of the genetic affinity among five geographically contiguous, linguistically and socio-culturally diverse Mongoloid-affiliated populations of Manipur in northeastern regions of India. SUBJECT AND METHODS: Blood samples were collected from unrelated and randomly selected volunteers of five ethnic populations (Meitei, Kuki, Naga, Hmar and Manipuri Muslim) from different parts of the state. Allelic variation in four minisatellite loci (D1S7, D4S139, D5S110 and D17S79) and three STR loci (vWA, FESFPS and F13AO1) was studied. RESULTS: Average heterozygosity values among the five groups for the minisatellite range from 68% to 94%, while the hypervariable three STR loci were between 60% and 88%. In the populations, all the studied loci were highly polymorphic, with almost no departure from Hardy-Weinberg equilibrium. The gene differentiation for the VNTR loci was lower and moderate (G(st) = 0.030) in comparison with microsatellites (G(st) = 0.043). The neighbour-joining method of clustering based on both type of molecular markers reveals a close cluster for the tribal groups of Kuki, Naga and Hmar, while Manipur Muslim stand distinct in both the trees. The clustering pattern obtained from the combined DNA marker loci matches more closely the pattern from STR loci than that obtained from VNTR loci. CONCLUSIONS: The results reinforce that using both VNTR and STR loci in detecting regional genetic affinity among the populations is more effective than using VNTR or STR independently, and also confirm the results obtained from the serological and electrophoretic data. However, the clustering pattern obtained from combined DNA markers is more in conformity with the pattern obtained by STR loci rather than with VNTR loci. Despite linguistic, geographical and cultural barriers, the populations show genetic affinity among the four populations except in the case of the migrant Manipur Muslim group. PMID- 12637189 TI - Sensitivity and specificity of body mass index and skinfold thicknesses in detecting excess adiposity in children aged 8-12 years. AB - PRIMARY OBJECTIVE: The study aimed to evaluate the sensitivity (SN) and specificity (SP) of body mass index (BMI) and skinfold thicknesses in detecting excess adiposity in children. RESEARCH DESIGN: Cross-sectional. MATERIALS AND METHODS: 986 children (500 females and 486 males) aged 10 +/- 1 years (mean +/- SD; range: 8-12 years) were studied. All underwent anthropometric measurements and bioelectrical impedance analysis (BIA). Dual-energy X-ray absorptiometry (DXA) was performed in 52 children to develop a population-specific algorithm for the assessment of fat-free mass (FFM) from BIA. The algorithm was applied to the remaining 934 children to estimate their FFM. Fat mass (FM) was obtained by subtracting FFM from weight (Wt). Values of FM:Wt were transformed in Z-scores and converted into 19 percentile categories (from 5 to 95 in steps of 5). The same procedure was performed with BMI and the log-transformed sum of four skinfold thicknesses (triceps, biceps, subscapular and suprailiac; lt-4SF). Excess adiposity was defined as a level of FM:Wt greater than the internally derived 85th percentile. SN and SP of each internally derived percentile of BMI and lt-4SF in detecting excess adiposity were calculated. RESULTS: In the pooled sample (n = 934), SN and SP were 0.39 and 0.99 for the 95th percentile of BMI, 0.65 and 0.95 for the 85th percentile of BMI, and 0.75 and 0.94 for the 85th percentile of lt-4SF. CONCLUSIONS: BMI percentiles employed in the present study have a high SP but a low SN in detecting excess adiposity in 8-12-year-old children. The use of the sum of four skinfolds has the potential to increase the SN of a screening programme for excess adiposity in children of this age. PMID- 12637190 TI - Consanguinity and genetic morbidity in a British primary care setting: a pilot study with trained linkworkers. AB - OBJECTIVES: To assess the feasibility of detecting consanguineous relationships and significant genetic morbidity through screening pedigrees administered by a primary care trained linkworker. SUBJECTS AND METHODS: A case-controlled crossover study with pedigree recording by both genetic nurse specialists and a primary care worker. From 1012 records of British Pakistani patients registered with an inner city practice, 14 women, identified as having increased genetic risk, were recruited (Group 1). A further 14 age- and parity-matched women, with no indication of genetic morbidity in their General Practice records (Group 2), were also recruited. RESULTS: Valuable genetic information, not recorded in General Practice records, were ascertained through the screening pedigrees, in four members of Group 1 (29%) and six members of Group 2 (43%). There was poor agreement between the coefficients of inbreeding recorded from pedigrees prepared by the primary care worker and genetic nurse specialists (Kappa = 0.157; 95% CI 0.028-0.286). CONCLUSION: Pedigrees can be utilized as a General Practice screening tool to detect relevant genetic morbidity, not ascertained in General Practice records. The process is enhanced when a primary care worker, from the same culture as the volunteers, collects information using the patients' mother tongue. PMID- 12637191 TI - Age at menarche in relation to anthropometric characteristics, competition level and boat category in elite junior rowers. AB - BACKGROUND: Within the context of the effects of training for sports on growth and maturation, there is very little menarcheal data for elite rowing athletes. Knowledge of the relationship of the maturational status with training level, different boat categories, and somatic features of the athletes will clarify the assumed impact of rowing training on the growth and maturational process of youngsters. AIM: The aim of this study was to determine the age at menarche in world top junior rowing athletes and to investigate its relationship with anthropometric characteristics, and competition level, rowing style and boat category. SUBJECTS AND METHODS: The sample consisted of 212 female junior rowers, with a mean chronological age of 17.6 +/- 0.8 years, all participants at the 1997 FISA World Junior Rowing Championships. Anthropometric dimensions, somatotype and body composition characteristics were measured, and age at menarche and training data were retrospectively obtained by questionnaires. RESULTS: Results revealed that the mean age at menarche of the total group of rowers was 12.8 +/- 1.2 years and did not differ from a non-athletic reference population. Rowers who started their rowing training before menarche (n = 78) showed a significant (p 0.05). CONCLUSION: Although a statistical significant sex effect is observed, it is less than the error of the measurement, and thus sex difference are debatable. In general, sex difference are explained by anthropometric difference. PMID- 12637193 TI - Genetic and environmental sources on familial transmission in Basque families. II. Stature, weight and body mass index. AB - PRIMARY OBJECTIVE: The present research analyses a cross-sectional sample of 359 individuals (107 fathers, 107 mothers, 71 sons and 74 daughters) in 107 nuclear families from the Basque Country (Spain). The aim was the estimation of both transmissible and non-transmissible components acting on familial resemblance for stature, weight and body mass index (BMI). METHODS: Standardized data of each generation and sex were adjusted to a BETA model of path analysis independent of sex effects, allowing the possibility of differentiating between transmissible (genetic and cultural) and non-transmissible (environmental) components acting on the observed phenotypic variance in Biscayan offspring. RESULTS: The results supported the full model of familial transmission for the three variables, in addition to other reduced models that gave rise to transmission effects regardless of any influence from the siblings' shared environment on the environmental transmitted component from their parents (b = 0), and without genetic effects on the phenotype (h = z = 0). The most parsimonious model was accepted for weight and rejected for stature and BMI. The correlations existing between children and their mothers were higher than those with their fathers. CONCLUSION: The results confirmed the hypothesis of maternal effects influencing the inheritance of stature. PMID- 12637194 TI - Sizes and obesity pattern of South Iranian adolescent females. AB - BACKGROUND: Sizes (height and weight) and obesity (a scaled weight-by-height index) charts of a representative sample of 1,743 healthy adolescent females of Shiraz (Southern Iran) aged 11-17 years are presented. METHODS: An adjusted weight-for-height was used to define a possible obesity index. Polynomial modelling was used by applying the HRY (Healy, Rasbash, Yang) nonparametric method to estimate age-related smoothed centiles of sizes and obesity. RESULTS: A ponderal index in the form of weight/height(3) represented obesity better than any other index which is logically related to weight/volume and enjoys biological justification. No more than cubic polynomials were needed to fit height-for-age, weight-for-age, obesity-for-age and weight-for-height smoothly. The 10th, 75th and 97th centiles of height and weight of our subjects lie on the 3rd, median and 90th centiles of the NCHS standard, respectively. Obesity pattern increases with age, giving an appropriate index to study obesity of female adolescents. However, weight-for-height chart independent of age range of subjects may serve as an alternative. CONCLUSION: It is concluded that the ponderal index is an appropriate index to study obesity of adolescent females, and is a simple one that is biologically plausible. However, other indices such as weight-for-height may be considered as an alternative. A local standard for assessing sizes and obesity of adolescent females is recommended for clinical as well as community health purposes in Iran. PMID- 12637196 TI - A randomized, controlled lower leg growth study of vitamin D supplementation to healthy children during the winter season. AB - BACKGROUND: The deceleration of longitudinal growth in children during winter occurs simultaneously with a decrease in the number of daylight hours and a reduction in vitamin D status. Due to worries about deleterious effects on bone of a relative insufficiency, vitamin D supplementation to healthy children has been suggested. AIM: To see whether supplementation of vitamin D to healthy children during winter affects seasonal growth. SUBJECTS AND METHODS: Twelve girls and eight boys aged 6.2-13.7 (mean 9.8) years, all healthy, were enrolled in a double-blind, randomized, placebo-controlled cross-over study with two 4 week treatment periods and 2-week run-in and wash-out periods. Vitamin D(3) 600 IU was given in one tablet ABCDin daily. Knemometry of the right lower leg was performed on the first and last day of each period. RESULTS: Lower leg growth rates (mean +/- SEM) during placebo and vitamin D(3) administration were identical: 0.28 +/- 0.04 mm per week (p = 0.94, t = 0.1, 95% CI: - 0.12-0.13 mm per week). CONCLUSION: Supplementation with vitamin D(3) 600 IU day(-1) to healthy children during winter may not improve seasonal growth. Therefore, supplementation may not be recommended on the grounds of concerns about growth; however firm conclusions await randomized long-term studies. PMID- 12637195 TI - Overweight and obesity in children and adolescents: relationship with blood pressure, and physical activity. AB - OBJECTIVE: Using body mass index (BMI) quartiles, the present study's aims were (1) to describe and evaluate the relationship between overweight/obesity with cardiovascular disease (CVD) risk factors, such as systolic blood pressure (SBP), diastolic blood pressure (DBP) and physical activity; (2) to provide information on the prevalence of high blood pressure and physical activity according to BMI quartiles; and (3) to examine the relationship of risk of obesity to adverse levels of high blood pressure and physical activity. METHODS: The sample consisted of 1439 children (758 males and 681 females), 8-16 years old. Participants were classified as being at 'risk of obesity' according to age- and sex-adjusted fourth quartile of BMI values. Blood pressure was measured using the Dinamap monitor. An adapted version of the Weekly Activity Checklist was applied for the diagnosis of physical activity index (PAI). RESULTS: Systolic and diastolic blood pressures were significantly (p < 0.05) and positively related to BMI, whereas no significant associations were reported between BMI and PAI in both genders. Overall, the proportion of subjects with adverse levels of blood pressure (SBP or DBP) ranged from 21.3% in the first BMI quartile (BMI-1Q) to 30.4% in BMI-4Q (p < 0.05). The PAI ranged from 24.5% in BMI-1Q to 20.6 in BMI-4Q and no significant differences were found. Children and adolescents at 'risk of obesity' were 1.5 times as likely to have at least one risk factor (high blood pressure or low level of activity). CONCLUSIONS: Our data suggest that higher BMI is associated with higher values of SBP and DBP. Children and adolescents in the upper quartile of BMI are 1.5 times as likely to have at least one risk factor. Furthermore our data confirmed previous findings that higher levels of BMI are associated with a unfavourable risk profile for CVD risk factors. PMID- 12637197 TI - ABO blood group in Amerindians from Brazilian Amazon. AB - BACKGROUND: The Parakana is a group of Indians with cultural similarities to the extinct Tupi group. They are an isolated native population from East Brazilian Amazon. A number of different O alleles have been found at the blood group ABO locus in populations of several ethnic origins (Caucasians, Blacks, Amerindians). AIM: The present study describes the ABO blood group polymorphism gene of the Parakana Indians. The Amerindian group was carefully selected for racial background. SUBJECT AND METHODS: The blood group polymorphism was analysed in genomic DNA from 62 Parakana Indians. We determined the 261G deletion, the T646A and C771T mutations described in O(1variant) and the G542A substitution, using PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism). RESULTS: All Amerindians studied were homozygous for the 261G deletion. The frequencies of the T646A and C771T mutations in Parakanas (0.65) were lower than that observed in Kayapo, Yanomama and Arara Indians (0.91) (chi (2) = 18.24; p-v < 0.001. The G542A substitution in Parakanas was also lower (0.22) than in other tribes (0.42) (chi(2) = 9.73; p-v = 0.001). CONCLUSIONS: The different O alleles including the G542A mutation are not distributed homogeneously among all Amazonian Amerindians. Our results are in agreement with other genetic markers studied previously in Parakana Indians, whose distinct genetic pattern differs from Europeans and even from other Amerindians. PMID- 12637201 TI - Brain-immune cross-talk. PMID- 12637198 TI - Height and secular trend in conscripts born in the Central Apennines (Italy), 1865-1972. AB - OBJECTIVE: The main aim of the research was to evaluate the effects of prolonged isolation on the secular trend of height in an internal zone of the central Apennines (Abruzzo, Province of L'Aquila). METHODS: Personal data for 1006 military conscripts in five sample years from 1865 to 1972 were used. The mean, standard deviation, median, skewness and kurtosis were calculated. Normal distribution of the data was assessed by the Kolmogorov-Smirnov test (Monte Carlo method). Differences between the means were tested by ANOVA and the Tukey test. RESULTS: Mean height increased from 161.3 +/- 5.97 cm in 1865 to 172.6 +/- 6.28 cm in 1972. The increases were slight, but constant, in the first 80 years: between 0.44 and 0.86 cm/decade, but greater thereafter: 2.37 cm/decade between 1945 and 1972. Regarding the frequency distribution, skewness showed negative values only in 1865 (-0.37 +/- 0.21). The Kolmogorov-Smirnov test did not show significance for any period. CONCLUSIONS: The results confirm that the mean heights in south-central Italy, including Abruzzo, were lower than those of the more economically advanced regions of northern Italy in each period. The difference has decreased greatly in the last few generations, but the secular trend is continuing in these communities. This seems to be due to prolonged isolation which has retarded economic development and favoured endogamy and genetic drift. However, it has also prevented the negative effects of wars, since there was no reduction of the secular trend in those periods. PMID- 12637202 TI - Cytokine stress responses depend on lateralization in mice. AB - In mice, lateralization as assessed by paw preference represents a behavioral trait linked to immune reactivity and stress susceptibility. Right-pawed mice are more reactive to stress than left-pawed animals when brain metabolism, activation of the corticoid axis, and depression of lymphoproliferation are studied. Since stress responses include cytokine production, we address the possibility that lateralization influences the production of cytokines--especially interleukin (IL)-1--responsible for depression of lymphoproliferation and activation of the corticoid axis. Increased plasma IL-1 level that may be considered as a stress marker, was observed in right- but not in left-pawed mice submitted to a 4 h restraint. Likewise, plasma levels were greater in right- than in left-pawed animals 2 h after the administration of a low dose of lipopolysaccharides (LPS). By contrast, there was no lateralization effect in restraint-induced plasma level of IL-6 or in the LPS-induced increase in plasma IL-10. Prazosin, an alpha1/alpha2 adrenoreceptor antagonist, drastically increased plasma IL-10 induced by LPS, reduced plasma levels of IL-1 and abolished the effect of lateralization observed after LPS alone. This suggests that alpha-adrenergic modulation of IL-1 production depends on lateralization through mechanisms that need further investigation. PMID- 12637203 TI - Neuroimmune stress responses: reciprocal connections between the hypothalamus and the brainstem. AB - Hypothalamic nuclei, particularly the paraventricular nuclei (PVN), are important brain sites responsible for eliciting stress responses following a systemic immune challenge. The activation of PVN cells by a systemic immune challenge is critically dependent on the integrity of inputs from brainstem cells situated in the nucleus tractus solitarius (NTS) and ventrolateral medulla (VLM). Interestingly, a descending pathway from the PVN to the brainstem, recruited by systemic immune challenge, might also exist. It is well documented that PVN neurons innervate the NTS and VLM and recent evidence from our laboratory shows that lesions of the PVN reduce brainstem cell responses elicited by a systemic bolus of the proinflammatory cytokine interleukin-1beta (IL-1beta). Although a number of different PVN divisions are candidates for the source of inputs to the brainstem, we have demonstrated that the majority of descending PVN projections recruited by systemic IL-1beta arise from cell bodies localized in the medial and lateral parvocellular PVN. These findings suggest that central nervous system responses to an immune challenge are likely to involve complex reciprocal connections between the PVN and the brainstem, whereby brainstem cell populations could essentially act as integratory sites for descending and ascending immune signals. For instance, these brainstem pathways may have significant implications not only for the regulation of central hypothalamic and extra-hypothalamic targets but also the autonomic nervous system. PMID- 12637204 TI - Stress and cytokine-elicited neuroendocrine and neurotransmitter sensitization: implications for depressive illness. AB - Stressful events, by their effects on neurotransmitter and neuroendocrine processes, are thought to favor the development or exacerbation of depressive illness. In as much as immunological challenge, may provoke stressor-like neuroendocrine and central neurochemical changes, the view was offered that immune activation essentially acts like a stressor and may contribute to the evolution of affective illness. In this respect, viral and bacterial infections appear to influence behavioral/metabolic (e.g. fever, anorexia, somnolence) and neurotransmitter functioning through the release of cytokines, which act as messengers between the immune system and brain. The present report provides a brief overview of the neurochemical consequences of proinflammatory cytokine treatments, particularly the actions of interleukin (IL)-1beta and tumor necrosis factor-alpha. As well, synergy with psychogenic and neurogenic stressors are described, as are data showing that cytokines, like stressors, may have time dependent proactive (sensitization) effects, so that reexposure to the treatments greatly augments hypothalamic-pituitary-adrenal activity, as well as central neurochemical changes. Indeed, the neurotransmitter alterations are not restricted to hypothalamic nuclei, but occur in several extrahypothalamic sites, including various limbic regions. It is suggested that by virtue of these neurochemical changes, cytokines may have both immediate and proactive effects on mood states. PMID- 12637205 TI - Differential effects of physical and psychological stressors on immune functions of rats. AB - To study the effects of different types or durations of stressors on immune functions, male Fischer rats were exposed to chronic physical (electric foot shock) or psychological (non-foot shock) stress induced in the communication box. Superoxide production by alveolar macrophages (AMs), mitogen-induced splenic lymphocyte proliferation, and splenic natural killer (NK) cell cytolysis were examined in vitro. Repeated exposure to physical stress suppressed superoxide production by AMs (-58%, p<0.05 for opsonized zymosan (OZ) and -51%, p<0.05 for phorbol 12-myristate 13-acetate (PMA)), although psychological stress suppressed superoxide production after 24 h of repeated exposures (-40%, p<0.05 for OZ and 47%, p<0.05 for PMA). Acute suppression of the blastic response of splenic lymphocytes was only found in the physical stress group (p<0.05), although the chronic effects were only found in the psychological stress group (p<0.05). NK cell activity was suppressed immediately after the acute physical stress (-30%, p<0.05), but no effects were found in the psychological stress group. These results underline the importance of distinguishing between physical versus psychological stressors when examining the effects of stress on immune functions. PMID- 12637206 TI - Observing a fictitious stressful event: haematological changes, including circulating leukocyte activation. AB - The aim of this study was to assess the effect of watching a psychological stressful event on the activation of leukocytes in healthy human volunteers. Blood samples were obtained from 32 healthy male and female subjects aged between 20 and 26 years before, during and after either watching an 83-minute horror film that none of the subjects had previously seen (The Texas Chainsaw Massacre, 1974) or by sitting quietly in a room (control group). Total differential cell counts, leukocyte activation as measured by the nitroblue tetrazolium (NBT) test, heart rate and blood pressure (BP) measurements were taken at defined time points. There were significant increases in peripheral circulating leukocytes, the number of activated circulating leukocytes, haemoglobin (Hb) concentration and haematocrit (Hct) in response to the stressor. These were accompanied by significant increases in heart rate, systolic and diastolic BP (P<0.05 from baseline). This is the first reported study on the effects of observing a psychologically stressful, albeit fictitious event on circulating leukocyte numbers and the state of leukocyte activation as determined by the nitrotetrazolium test. PMID- 12637207 TI - Proactive sensitizing effects of acute stress on acoustic startle responses and experimentally induced colitis in rats: relationship to corticosterone. AB - In humans, some individuals develop a syndrome after trauma (post-traumatic stress disorder, PTSD) characterized by increased startle responses and lower than normal cortisol secretion. We explored a rat model using the acoustic startle response (ASR) as a behavioral indicator of the effect of a short series of shocks. Because gastrointestinal disorders have been associated with prior stress, we also studied the rats' vulnerability to a chemically-induced colitis. After initial blood sampling, 12 rats were exposed to ten 1 mA 5 s foot-shocks while 12 rats served as controls. Nineteen days later the rats were tested for ASR. Thirty trials (10 trials at each of 95, 105, and 115 dB, pseudo-randomized) were given. After exposure for 6 days to dextran sulphate sodium in their drinking water, the rats were killed and the colons examined for erosions. Shocked rats showed greater startle responses and more colonic erosion than unshocked rats, but the shock effects were significant only for animals with low initial plasma corticosterone levels. Shocked rats also showed higher levels of granulocyte marker protein (GMP) in their faeces. These results suggest that low corticosterone secretion may represent a marker for vulnerability to long term effects of shocks as indicated by increased startle responses and colonic pathology. PMID- 12637208 TI - Restraint stress and corticotropin releasing hormone modulation of murine cutaneous POMC mRNA. AB - The skin is a unique immunological defense barrier that protects the organism from occupational and environmental exposures and provides a model system in which to evaluate the interaction of the central nervous system with the peripheral immune response. In the studies presented here, we tested mild, acute restraint stress activation of the cutaneous corticotropin releasing hormone-pro opiomelanocortin (CRH-POMC) axis. We verified that 2 h restraint stress increased the serum concentration of corticosterone and alpha-melanocyte stimulating hormone. We report for the first time that CRH upregulates POMC mRNA expression in mouse skin in vitro. We also demonstrated, by RT-PCR, that 2,4 di nitrofluorobenzene (DNFB) upregulates cutaneous POMC mRNA expression, the production of which is suppressed by restraint stress. These data confirm the presence and functionality of two hormones of the hypothalamo-pituitary axis in the skin and suggest that activation of the central hypothalamo-pituitary-adrenal axis may over ride activation of the cutaneous CRH-POMC mechanism in the development of DNFB-stimulated allergic contact dermatitis. PMID- 12637209 TI - Repeated acute stress alters heart morphometry in male and female rats differently. AB - Stress exerts deleterious effects on the cardiovascular system. Left ventricular hypertrophy has been identified as a risk factor for heart disease. The effects of stress on other heart parameters, including heart size and heart shape, are not well-characterized. In addition, the extent to which males and females differ in stress effects on these measures has not been examined, but may help to further explain gender differences in heart disease morbidity and mortality. The present experiment examined effects of mild daily stress on heart morphology and blood volume (obtained by ex-sanguination at decapitation) in 39 male and 40 female rats of two strains. Following 14 days of stress exposure, rats were killed and blood volume was measured. Heart length, heart weight, left ventricle cavity width, right ventricle width, lateral wall thickness, anterior wall thickness, posterior wall thickness, and septal wall thickness were measured. In males, after controlling for body weight, stress decreased heart length and left ventricle cavity width and increased septal wall thickness. In females, after controlling for body weight, stress reduced total blood volume, but did not significantly affect other heart parameters. These findings suggest that stress exposure can alter heart morphology and blood volume and that there are sex differences in these effects. PMID- 12637210 TI - Brain-immune interactions in stress--the impact of hormones on disease. PMID- 12637214 TI - New antileukemic agents. AB - Despite the tremendous progress in the treatment of childhood leukemias over the last 50 years, certain subgroups of children continue to have poor prognosis. Hence, there is a need for development of new antileukemic agents. In this review, the authors describe results of clinical trials of several new antileukemic compounds with different mechanisms of action (signal transduction inhibitors, nucleoside analogs, DNA hypomethylators, angiogenesis inhibitors, and monoclonal antibodies). Although most of these compounds are not used in pediatric leukemias, the concepts surrounding their clinical development are important to all pediatric hematologists/oncologists. PMID- 12637215 TI - Methotrexate infusions as central nervous system prophylaxis in children with acute lymphocytic leukemia: the Norwegian experience. AB - This study included all 690 children in Norway diagnosed as having acute lymphocytic leukemia (ALL) from July 1975 till the end of 1997. Relapses and deaths were monitored until the end of 2000. Neuroleukemia prophylaxis was intravenous methotrexate (MTX) infusions as intermediate-dose methotrexate (IDM) or high-dose methotrexate (HDM) combined with intrathecal MTX. From 1992, systemic therapy was considerably intensified, and, in addition, patients in a subgroup of the high-risk and very high-risk groups were given prophylactic cranial irradiation. The overall findings showed that MTX significantly reduced central nervous system (CNS)-related relapses, and, in general, reinforced systemic therapy reduced significantly non-CNS relapses and deaths. The overall crude survival was 75%. During the study period, the crude survival improved for patients on standard protocols from initially 65 to 90%. Forty patients (6%) developed isolated CNS relapse, 27 (4%) had combined CNS relapse, whereas 180 (26%) had non-CNS relapse. When IDM and HDM were compared, the cumulative risk for isolated CNS relapse was significantly lower with HDM, 12 and 5%, respectively. For any relapses that involved the CNS, the risk remained significantly lower for HDM, 8 versus 18%. Of the 40 patients with isolated CNS relapse, 23 survived (58%). PMID- 12637216 TI - Possible benefits of high-dose chemotherapy as intensive consolidation in patients with high-risk rhabdomyosarcoma who achieve complete remission with conventional chemotherapy. AB - The authors reviewed their single-center experience with autologous stem cell transplantation (SCT) in 22 patients with advanced rhabdomyosarcoma. Pathological subtypes included alveolar (n = 7) and embryonal types (n = 15). The conditioning regimen primarily consisted of etoposide, carboplatin, and melphalan. Fourteen, five, and three patients underwent SCT in CR, PR, and PD, respectively. Eight patients are currently alive without evidence of disease. The overall survival rate at 5 years was 70% for 14 patients who were in CR at the time of SCT. This limited experience warrants the examination of SCT in a prospective study. PMID- 12637217 TI - Experience of the Izmir Pediatric Oncology Group on Neuroblastoma: IPOG-NBL-92 Protocol. AB - This multicentric study aimed to bring neuroblastoma patients together under IPOG NBL-92 protocol and evaluate the results within the period between 1992 and 2001 in Izmir. Sixty-seven neuroblastoma patients from 4 pediatric oncology centers in Izmir were included in the study. IPOG-NBL-92 protocol modified from German Pediatric Oncology (GPO)-NB-90 protocol was applied: Patients in stage 1 received only surgery, while surgery plus 4 chemotherapy courses (cisplatin, vincristine, ifosfamide) were given in stage 2 and surgery plus 6 chemotherapy courses (cisplatin, vincristine, ifosfamide, epirubicin, cyclophosphamide) were given in stages 3 and 4 patients. In patients who were kept in complete remission (CR), a maintenance therapy of one year was applied. Radiotherapy was given to the primary site following induction chemotherapy plus surgery in stages 3 and 4 patients with partial remission (PR). The stages of the patients were as follows: 5% in stage 1, 39% in stage 3, 49% in stage 4, and 7% in stage 4S. Primary tumor site was abdomen in 88% of cases. CR rates were as 100% in stage 1, 76% in stage 3, 35% in stage 4, and 75% in stage 4S. Relapse was observed in 32% of patients in a median of 19 months. The median follow-up time for survivors was 33 (17-102) months. Five-year OS rate was 31% and the EFS rate was 30% in all patients. Five year overall and event-free survival rates were 63 and 30% in stage 3, but 6 and 5%, respectively, in stage 4 patients. Univariate analysis established that the age, stage, primary tumor site, and high LDH and NSE levels conferred a significant difference. The IPOG-NBL-92 protocol has proved to be satisfactory with tolerable toxicity and reasonable CR and survival rates. However, more effective treatments suitable to Turkey's social and economic conditions are urgently needed for children over 1 year of age with advanced neuroblastoma. PMID- 12637218 TI - Thrombophilia in infancy: factor V Leiden and MTHFR or factor II double heterozygocity as a risk factor. AB - Thrombophilic risk factors are associated with thromboembolism in children but data in infants and neonates are not well established. The authors report a series of 9 infants with thrombotic events and the associated genetic risk factors. The clinical and laboratory records of newborns and infants with a history of thrombotic events were summarized, while patients with underlying diseases were excluded. The frequency of the genetic mutations was compared to a control group of 80 children from the same ethnic origin. In 6 patients a cerebrovascular accident was diagnosed and in 3 newborns, CT scan could diagnose antenatal brain infarct. In another 2 patients deep-vein thrombosis associated with femoral catheterization was diagnosed. Seven infants were factor V Leiden heterozygous and another one homozygous. Methylenetetrahydrofolate reductase genotype was found in 5 infants. Five cases were found to be double heterozygous for those two mutations, and another one double heterozygous for FVL and factor II. The results of this small series of patients indicate that cerebrovascular accident is the major thrombotic event in infants and the combination of more than one prothrombotic factors may be the cause of those events. The correct management, including anticoagulant therapy, is still under discussion and waiting for larger series and long-term follow-up results until accurate recommendations can be made. PMID- 12637219 TI - Liver ferritin subunit ratios in neonatal hemochromatosis. AB - Neonatal hemochromatosis is an enigmatic disease. Little is known about iron metabolism in this disease, including the tissue concentration of ferritin or its H and L subunit ratio. The authors report the tissue iron, ferritin, and ferritin subunit content of a child who died at 5 weeks of neonatal hemochromatosis. The child was born at 29 weeks gestation to a mother with lupus, sickle cell trait, and gestational diabetes. The child's severe liver dysfunction led to the clinical diagnosis of neonatal hemochromatosis at 1 week of age. Despite aggressive support, including red cell transfusions and chelation, the child died of an intracranial hemorrhage. Autopsy showed liver fibrosis and iron staining characteristic of neonatal hemochromatosis. Autopsy liver tissue was compared to age-matched control tissue. Soluble protein was analyzed by the Bradford method. Soluble iron (over 90% of total iron) was analyzed by the o-phenanthroline complex. Tissue ferritin and human ferritin controls (Calzyme) were analyzed by Western blotting after SDS-PAGE, identified with sheep anti-human ferritin antibodies (The BindingSite) secondary antibody-fluorescence for detection, and quantified using the Molecular Dynamics Storm 840 phosphorimager and ImageQuant software. Protein, iron, and total ferritin were similar in the normal and neonatal hemochromatosis liver tissues. Ferritin subunits, however, showed an increased H/L-subunit ratio compared to an age-matched control. This first report of a marked increase in the ferritin H/L-subunit ratio may point to an underlying mechanism of disease in this enigmatic disorder. PMID- 12637221 TI - Non-Hodgkin lymphoma in a child presenting with acute paraplegia: a case report. AB - Spinal cord compression is a rare presentation of non-Hodgkin lymphoma. Extradural location at onset is a rare but devastating event in pediatric oncology. The authors describe a girl with acute spinal cord compression due to epidural non-Hodgkin lymphoma, emphasizing the encouraging perspective for a complete recovery in children with this condition. A 5-year-old girl presented with pain followed by progressive hyposthenia and paraplegia after a trauma. CT scan and MRI showed homogeneous tissue extending from T2 to L4, occupying the entire vertebral canal and extending to the para- and peri-vertebral soft parts. Emergency surgical debulking was carried out through T6-L1 laminectomy. The patient began chemotherapy (LMB 89 Protocol) and the tumor quickly disappeared. The patient is maintaining a complete remission 42 months after diagnosis. Significant results may be obtained with the chemotherapy treatment of epidural non-Hodgkin lymphoma when the disease is promptly diagnosed. Considering the effectiveness of chemotherapy, the authors believe that a neuro-surgical approach should be employed only when rapid worsening of symptoms is observed or for diagnostic purpose. PMID- 12637220 TI - Neutrophil apoptosis in patients with beta-thalassemia major. AB - Increased susceptibility to infection is reported in patients with beta thalassemia major due to toxic effect of iron on neutrophil functions and reticuloendothelial system dysfunction. This study investigated the association between the neutrophil apoptosis and frequency of infection episodes, desferrioxamine treatment, and serum ferritin levels in patients with beta thalassemia major. A total of 35 children diagnosed with beta-thalassemia major were enrolled. Group 1 consisted patients who were receiving desferrioxamine (DFO) and group 2 consisted of patients who did not start to receive DFO. A total of 15 healthy children were enrolled to serve as a control group. Frequency of infection episodes within a year was noted from hospital records. In all patients, the same method based on flow cytometry (annexin V labeled with FITC) was used to assess neutrophil apoptosis. Neutrophil count and percentage of apoptotic neutrophils did not differ significantly between the groups. When frequency of infection episodes among groups was evaluated, frequency of infection episodes of the patients who were receiving DFO was significantly higher than in the other groups. When correlation between neutrophil apoptosis and frequency of infection episodes, serum ferritin levels, and neutrophil count of the patients was analyzed according to groups, no significant correlation was found. The results indicate that high serum ferritin level and DFO use in patients with beta-thalassemia major do not enhance neutrophil apotosis in vivo and enhanced neutrophil apoptosis cannot be a possible cause for increased susceptibility to infections in these patients. PMID- 12637222 TI - Amifostine protects against cisplatin-induced nephrotoxicity in a child with medulloblastoma. PMID- 12637223 TI - Ureteral obstruction following allogeneic bone marrow transplantation in children. AB - Two patients with Ph + CML underwent URD-BMT after conditioning with Bu-Cy-LPAM. They developed hemorrhagic cystitis with an extremely complicated and painful course, caused by ureteral obstruction, requiring prolonged hospitalization. No virus other than cytomegalovirus was found and in both cases was attributed to Cy use. Treatment is usually conservative, but in the case of severe obstruction, a surgical approach should be considered and performed as early as possible to preserve renal function. PMID- 12637224 TI - Association of wilms tumor with spinal dysraphism. AB - Previous data suggested an association of vertebral anomalies with Wilms tumor. At the same time, vertebral midline fusion defects are often indicated by dermal anomalies over the spine. In the present study the prevalence of both occult spina bifida and cutaneous signs of spinal dysraphism was significantly higher in 50 Wilms patients than in 180 control children (18.0 versus 4.4%, p <.01, and 35.9 versus 17.5%, p <.02, respectively). Family investigations are needed to answer the question whether signs of spinal dysraphism in parents and sibs of patients may be regarded as indicators of an increased risk of Wilms tumor in the family. PMID- 12637225 TI - Transforming growth factor-beta promotes fibroblast apoptosis induced by H2O2. AB - Transforming growth factor-beta (TGF-beta) is believed to play a central role in fibroblastic wound repair and subsequent scar formation. Fibroblasts undergo apoptosis as granulation tissue evolves into a scar. To determine whether TGF beta influences fibroblast apoptosis, human lung fibroblasts (IMR-90) were treated with TGF-beta (2 to 10 ng/mL) and then exposed to H(2)O(2) (50 to 150 microM), an inducer of fibroblast apoptosis. Apoptosis was evaluated by nuclear staining with Hoechst33342 and terminal deoxynucleotidyl transferase (TdT) mediated nucleotide nick-end labeling. TGF-beta alone did not induce fibroblast apoptosis, but it dose-dependently augmented the apoptosis induced by exposure to H(2)O(2). TGF-beta also increased the intracellular level of peroxides measured with carboxydichlorodihydrofluorescein, but it did not affect the protein levels of Bcl-2, Bax, Bad, or p53 proteins. These results suggest that TGF-beta promotes lung fibroblast apoptosis induced by H(2)O(2), probably by increasing intracellular peroxides. Thus, TGF-beta may promote the elimination of fibroblasts from wounds, particularly under conditions of exposure to enhanced oxidative stress in the lung. PMID- 12637226 TI - Distal pulmonary cell proliferation is associated with the expression of EIIIA+ fibronectin in the developing rat lung. AB - Fibronectin is known to regulate the development of the lung. The EIIIA segment of fibronectin is one of the major alternatively spliced segments and modulates the cell proliferative potential of fibronectin in vitro. But the specific role of the EIIIA-containing fibronectin isoform (referred to as EIIIA+ fibronectin) in pulmonary cell proliferation has not been explored in vivo as yet. In this study, to see whether EIIIA+ fibronectin is associated with pulmonary cell proliferation in vivo, the authors immunohistochemically examined the spatiotemporal relationship of the EIIIA+ fibronectin isoform protein and proliferating cell nuclear antigen (PCNA), a biochemical marker for the cell proliferation, during the alveolar formation in the developing rat lung. EIIIA+ fibronectin protein was localized in the epithelial cells and in the mesenchymal tissues. The expression of EIIIA+ fibronectin protein gradually decreased from the pseudoglandular stage to the saccular stage and then slightly increased from the saccular stage to the alveolar stage. This change in the EIIIA+ fibronectin expression seemed to be in accord with the change in the number of PCNA-positive cells in the distal pulmonary cells throughout the lung development. These results suggest that the expression of EIIIA+ fibronectin is associated with the distal pulmonary cell proliferation during the alveolar formation. PMID- 12637227 TI - Suppressive effects of diesel exhaust particles on cytokine release from human and murine alveolar macrophages. AB - Epidemiological studies have shown an increase in the number of hospital admissions for respiratory diseases in association with high concentrations of particulate matter smaller than 10 micro m (PM(10)). Diesel exhaust particles (DEP) are important components of PM(10). This study was designed to test the effect of DEP on the release of cytokines from alveolar macrophages (AMs). Human and murine AMs were exposed to DEP for 24 hours, and the concentrations of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and IL-8 were measured in the supernatants by enzyme-linked immunosorbent assay (ELISA). DEP (10 micro g/mL) suppressed the spontaneous release of TNF-alpha and IL-6 from murine AMs (P<.05). This suppression was not seen with exposure to carbon particles. Soluble components of DEP had a similar suppressive effect, suggesting that the chemical composition of DEP is responsible for the suppression. Lipopolysaccharide (LPS)- or IFN-gamma-induced TNF-alpha and IL-6 production by murine AMs were suppressed by DEP in a dose-dependent manner (P<.05). DEP also inhibited LPS-stimulated production of TNF-alpha, IL-6, and IL-8 from human AMs (P<.05). Pretreatment of AMs with superoxide dismutase (SOD) (300 IU/mL) prevented the suppressive effect of DEP on AM cytokine production (P<.05). The authors conclude that DEP exposure suppressed the release of cytokines from AMs, and speculate that this suppression could impair normal host defenses. PMID- 12637228 TI - Effects of neurokinins on airway and alveolar eosinophil recruitment. AB - The authors assessed the role of substance P (SP) and neurokinin A (NKA) and their receptor antagonists (RAs) SR140333 and SR48968 (respectively for NK(1) and NK(2) receptors) in pulmonary eosinophil influx induced by stimulation of capsaicin (CAP)-sensitive nerve terminals. The increase in respiratory system resistance after capsaicin infusion was attenuated by NK(2)RA and association of NK(1)NK(2)RA (P<.001). Respiratory system elastance (Ers) increase was attenuated with use of NK(1)NK(2)RA (P<.001). In alveolar wall, there was an increase in eosinophils after 30 minutes of CAP infusion (P<.001) and was attenuated after 24 hours. Pretreatment with NK(1)RA, NK(2)RA, and NK(1)NK(2)RA decreased eosinophils in alveolar wall (P<.001). SP induced an increase of eosinophils in alveolar wall (P<.001), although NKA may also contribute to this response. In airway wall, the authors observed an increase of eosinophils at 30 minutes (P=.006) till 24 hours after CAP infusion. They noticed a predominant influx of cells around airway wall after CAP and SP infusion. Pretreatment with NK(1)RA and NK(1)NK(2)RA reduced eosinophils (P<.001) in airway wall. Both SP and NKA contribute to eosinophil lung recruitment in distal airways and in alveolar wall, and these findings suggest that neurokinins may contribute to the development of eosinophilic inflammation in both allergic asthma and hypersensitivity pneumonitis. PMID- 12637229 TI - Combining histology and biochemical measurements of connective tissue components in small samples of lung: application to bleomycin-induced fibrosis in the mouse. AB - Bleomycin-induced lung injury in mice was used to illustrate the plausibility of quantitating changes in elastin or protein concentration in histologically defined regions of mouse lungs. Mice were instilled by either the intranasal or intratracheal route with varying levels of bleomycin and the lungs were removed and fixed under pressure for histology and analysis. Histological sections that bracketed a defined volume of tissue was used for biochemical analysis and was limited to the alveolar region of the lung. Our studies showed that the intranasal method of instillation was as effective as the intratracheal method for inducing fibrosis. Even though distribution to the 4 major lobes of the lung was fairly uniform, there was no consistency in the degree of fibrosis between the various lobes of the same animal. The most dramatic change in biochemical parameters was protein concentration, which increased over 10-fold in some fibrotic lungs compared to normal lungs, whereas desmosine concentration increased up to 5-fold. Fibrosis scores agreed well with desmosine concentration, except with moderate desmosine increases where histologically defined fibrosis was not observed. The experiments illustrate the feasibility and desirability of comparing histology and biochemical analysis on the same fixed lung sample. PMID- 12637230 TI - An ergonomic evaluation of snowmobiles. PMID- 12637231 TI - Measurement of biocides in metalworking fluids and in workplace air using capillary electrophoresis. PMID- 12637232 TI - 24-hour noise dose and risk assessment. PMID- 12637234 TI - Determination of known exhalation valve damage using a negative pressure user seal check method on full facepiece respirators. AB - A negative pressure user seal check (NPUSC) method was evaluated for its ability to adequately detect known exhalation valve leakage into a respirator. Three valves with different types of damage were included. Twenty-six test subjects, wearing full facepiece respirators, were asked to perform a NPUSC. Their responses as to whether they passed or failed the user seal check were compared to fit testing results from two quantitative fit test methods: ambient aerosol and controlled negative pressure. In addition, equipment developed at the University of Cincinnati was used to measure in-mask pressures that are generated during the performance of NPUSCs. This technique was employed to assess the ability of respirator wearers to properly conduct user seal checks. The data were analyzed to determine if the user seal check procedure is an effective method for detecting known exhalation valve damage. All test subjects reported passing the user seal check with the undamaged valve. With the warped valve installed, 95 percent of test subjects reported passing the user seal check. With the slit valve installed, 73 percent of test subjects reported passing. With the dirty valve installed, 65 percent reported passing. All fit factors, measured with the damaged valves, were below the Occupational Safety and Health Administration recognized pass/fail criteria except one fit test with the respirator equipped with the slit valve. Results from the in-mask pressure measurements confirmed whether or not the subject properly conducted a user seal check, but did not detect respirator leakage. In conclusion, the performance of a NPUSC rarely helped to identify damaged exhalation valves. These results support the need for respirator inspection prior to donning with periodic fit testing and the performance of user seal checks as necessary components of an adequate respiratory protection program. PMID- 12637235 TI - Numerical determination of personal aerosol sampler aspiration efficiency. AB - In this work the determination of the aspiration efficiency of personal aerosol samplers, commonly used in occupational exposure assessment, is investigated by means of CFD techniques. Specifically, it will be described a code to calculate the particle trajectories in a given flow field. At the present state the code considers only the effects of the mean flow field on the particle motion, whereas the turbulent diffusion effects are neglected. Comparisons with experimental measurements are also given in the framework of a research contract, supported by the European Community, with several experimental contributions from the participants. The main objective of the European research is to develop a new approach to experimentation with airborne particle flows, working on a reduced scale. This methodology has the advantage of allowing real-time aerosol determination and use of small wind tunnels, with a better experimental control. In this article we describe how the methodology has been verified using computational fluid dynamics. Experimental and numerical aspiration efficiencies have been compared and the influence of gravity and turbulence intensity in full and reduced scale has been investigated. The numerical techniques described here are in agreement with previous similar research and allow at least qualitative predictions of aspiration efficiency for real samplers, taking care of orientation from the incoming air flow. The major discrepancies among predicted and experimental results may be a consequence of bounce effects, which are very difficult to eliminate also by greasing the sampler surface. PMID- 12637237 TI - Engineering controls for selected silica and dust exposures in the construction industry--a review. AB - This literature review summarizes engineering control technology research for dust and silica exposures associated with selected tasks in the construction industry. Exposure to crystalline silica can cause silicosis and lung fibrosis, and evidence now links it with lung cancer. Of over 30 references identified and reviewed, 16 were particularly significant in providing data and analyses capable of documenting the efficacy of various engineering controls. These reports include information on generation rates and worker exposures to silica and dust during four different tasks: cutting brick and concrete block, grinding mortar from between bricks, drilling, and grinding concrete surfaces. The major controls are wet methods and local exhaust ventilation. The studies suggest that while the methods provide substantial exposure reductions, they may not reduce levels below the current ACGIH threshold limit value (TLV) of 0.05 mg/m(3) for respirable quartz. Although further research on controls for these operations is indicated, it is clear that effective methods exist for significant exposure reduction. PMID- 12637236 TI - Making sense of highway construction: a taxonomic framework for ergonomic exposure assessment and intervention research. AB - Construction is one of the most hazardous industries in the United States. Occupational health research to characterize the hazards in construction work has been hampered by the lack of a systematic approach to classification of construction work and its associated hazards. A taxonomy of construction work, a nested system of classification, has been developed to systematize the collection and reporting of exposure assessment data for the characterization and reduction of hazards and the prevention of musculoskeletal injury. This taxonomy subdivides construction work into the categories of stage, operation, task, and activity. It is based on a bidding specification system already in use within the industry and thus provides a terminology common among workers, supervisors, and managers. The identification of tasks and activities that are present in multiple stages and/or trades contributes to the efficiency of exposure data collection and facilitates the generalizability to other settings for both exposure data and intervention evaluations. The taxonomy provides a framework and vocabulary that facilitates field work and participatory research activities. It can also potentially be linked to personnel and economic data for estimation of costs of safety and health problems, as well as benefits of interventions. Although developed for construction ergonomics, the taxonomic approach has application to non-routine work in other industry sectors and possibly in occupational health research other than ergonomics. PMID- 12637238 TI - Pulmonary deposition modeling with airborne fiber exposure data: a study of workers manufacturing refractory ceramic fibers. AB - Increasing production of refractory ceramic fiber (RCF), a synthetic vitreous material with industrial applications (e.g., kiln insulation), has created interest in potential respiratory effects of exposure to airborne fibers during manufacturing. An ongoing study of RCF manufacturing workers in the United States has indicated an association between cumulative fiber exposure and pleural plaques. Fiber sizing data, obtained from electron microscopy analyses of 118 air samples collected in three independent studies over a 20-year period (1976-1995), were used with a computer deposition model to estimate pulmonary dose of fibers of specified dimensions for 652 former and current RCF production workers. Separate dose correction factors reflecting differences in fiber dimensions in six uniform job title groups were used with data on airborne fiber concentration and employment duration to calculate cumulative dose estimates for each worker. From review of the literature, critical dimensions (diameter <0.4 microm, length <10 microm) were defined for fibers that may translocate to the parietal pleura. Each of three continuous exposure/dose metrics analyzed in separate logistic regression models was significantly related to plaques, even after adjusting for possible past asbestos exposure: cumulative fiber exposure, chi(2) = 15.2 (p < 0.01); cumulative pulmonary dose (all fibers), chi(2) = 14.6 (p < 0.01); cumulative pulmonary dose (critical dimension fibers), chi(2) = 12.4 (p < 0.01). Odds ratios (ORs) were calculated for levels of each metric. Increasing ORs were statistically significant for the two highest dose levels of critical dimension fibers (level three, OR = 11, 95%CI = [1.4, 98]; level four, OR = 25, 95%CI = [3.2, 190]). Similar associations existed for all metrics after adjustment for possible asbestos exposure. It was concluded that development of pleural plaques follows exposure- and dose-response patterns, and that airborne fibers in RCF manufacturing facilities include those with critical dimensions associated with pleural plaque formation. Analysis of additional air samples may improve estimates of the dose-response relationship. PMID- 12637239 TI - Fatal injuries in the United States involving respirators, 1984-1995. AB - There is little published information concerning the epidemiology of fatal injuries involving respiratory protection. We compiled a case series from U.S. Occupational Safety and Health Administration investigation reports from 1984 through 1995. For the 12-year period there were 41 incidents resulting in 45 deaths due to asphyxiation or chemical poisoning while wearing a respirator. There were 23 deaths related to airline respirators, 17 deaths involving use of negative pressure (air purifying) respirators, and 5 deaths involving self contained breathing apparatus. Among the 23 deaths involving airline respirators, 15 were associated with compatible connection couplings for breathable air and inert gases. Three workers with beards died who wore tight-fitting respirators in an atmosphere that was immediately dangerous to life and health. Most of the fatalities involved regulatory and procedural violations, and would have been prevented by proper training and compliance with existing regulations. The information concerning the victims was limited but it did not appear that medical screening would have prevented any of the deaths. PMID- 12637242 TI - Homology modelling of human CYP1A2 based on the CYP2C5 crystallographic template structure. AB - 1. The results of homology modelling of human cytochrome P4501A2 (CYP1A2) based on the CYP2C5 crystal structure are reported. It exhibits improved sequence homology relative to that of CYP102. 2. It was demonstrated that many selective substrates for CYP1A2 could fit within the putative active site of the enzyme, and in orientations which agree with documented evidence for CYP1A2-mediated metabolism. 3. Furthermore, a number of amino acid residues lining the haem pocket have been shown, via site-directed mutagenesis, to have an influence on substrate metabolism, and these experimental findings from the literature are consistent with the modelled interactions for selective substrates. 4. The binding affinities of several CYP1A2 substrates have also been calculated from the CYP1A2 active site interactions and they agree closely with experimental values. PMID- 12637241 TI - mRNA and protein expression of dog liver cytochromes P450 in relation to the metabolism of human CYP2C substrates. AB - 1. Interpretation of novel drug exposure and toxicology data from the dog is tempered by our limited molecular and functional knowledge of dog cytochromes P450 (CYPs). The aim was to study the mRNA and protein expression of hepatic dog CYPs in relation to the metabolism of substrates of human CYP, particularly those of the CYP2C subfamily. 2. The rate of 7-hydroxylation of S-warfarin (CYP2C9 in humans) by dog liver microsomes (mean +/- SD from 12 (six male and six female) dogs = 10.8 +/- 1.9 fmol mg(-1) protein min(-1)) was 1.5-2 orders of magnitude lower than that in humans. 3. The rate of 4'-hydroxylation of S-mephenytoin, catalysed in humans by CYP2C19, was also low in dog liver (4.6 +/-1.5 pmol mg(-1) protein min(-1)) compared with human liver. In contrast, the rate of 4' hydroxylation of the R-enantiomer of mephenytoin by dog liver was much higher. The kinetics of this reaction (range of K(m) or K(0.5) 15-22 micro M, V(max) 35 59 pmol mg(-1) protein min(-1), n = 4 livers) were consistent with the involvement of a single enzyme. 4. In contrast to our findings for S-mephenytoin, dog liver microsomes 5'-hydroxylated omeprazole (also catalysed by CYP2C19 in humans) at considerably higher rates (range of K(m) 42-64 micro M, V(max) 22-46 pmol mg(-1) protein min(-1), n = 4 livers). 5. For all the substrates except omeprazole, a sex difference in their metabolism was observed in the dog (dextromethorphan N-demethylation: female range = 0.7-0.9, male = 0.4-0.8 nmol mg(-1) protein min(-1) (p < 0.02); S-warfarin 7-hydroxylation: female = 9-15.5, male = 8-12 fmol mg(-1) protein min(-1) (p < 0.02); R-mephenytoin 4' hydroxylation: female = 16-35, male = 11.5-19 pmol mg(-1) protein min(-1) (p < 0.01); omeprazole 5'-hydroxylation: female = 15-20, male 13-22 pmol mg(-1) protein min(-1) (p < 0.2)). 6. All dog livers expressed mRNA and CYP3A12, CYP2B11, CYP2C21 proteins, with no sex differences being found. Expression of CYP2C41 mRNA was undetectable in the livers of six of 11 dogs. 7. Correlation analysis suggested that CYP2B11 catalyses the N-demethylation of dextromethorphan (mediated in humans by CYP3A) and the 4'-hydroxylation of mephenytoin (mediated in humans by CYP2C19) in the dog, and that this enzyme and CYP3A12 contribute to S-warfarin 7-hydroxylation (mediated in humans by CYP2C9). 8. In conclusion, we have identified a distinct pattern of hepatic expression of the CYP2C41 gene in the Alderley Park beagle dog. Furthermore, marked differences in the metabolism of human CYP2C substrates were observed in this dog strain compared with humans with respect to rate of reaction, stereoselectivity and CYP enzyme selectivity. PMID- 12637243 TI - Hydrolysis by lactase phlorizin hydrolase is the first step in the uptake of daidzein glucosides by rat small intestine in vitro. AB - 1. Isoflavones are naturally occurring oestrogenic compounds found in plants, where they exist in the glycosylated form. A proportion of ingested glycosides appears to be absorbed in the upper gastrointestinal tract, where enterocytes play an important role in their metabolism. 2. One hypothesis is that ingestion may involve hydrolysis by the luminally exposed enzyme lactase phlorizin hydrolase (LPH), an enzyme expressed specifically at the small intestinal brush border. 3. Using an everted sac preparation of rat jejunum and an inhibitor of LPH, we investigated the absorption of daidzein-O(7)-glucoside (daidzin) and the effect of LPH inhibition on this process. It was demonstrated that LPH plays a major role in the deglycosylation of daidzin. 4. The hydrolysis product, daidzein, is absorbed by epithelial cells and glucuronidated to daidzein-O(7) glucuronide, which is subsequently exported primarily to the serosal (vascular) side of the tissue rather than to the luminal side. 5. A small but significant proportion of the intact glycoside is also transferred to the serosal compartment, and in the presence of an LPH inhibitor this was enhanced with a corresponding reduction in deglucosylation and glucuronidation. 6. The results indicate that that LPH plays an important role in the metabolism of glycosylated phytochemicals, and that the expression and activity of this enzyme in the small intestine can modify the profile of metabolites appearing in the circulation. PMID- 12637244 TI - Characterization of the cytochrome P450 enzymes involved in the in vitro metabolism of ethosuximide by human hepatic microsomal enzymes. AB - 1. The relative roles of human hepatic cytochrome P450 (CYP) subfamilies participating in ethosuximide metabolism have been studied in vitro using humanized heterologous CYP microsomal systems expressing either CYP1A2, CYP2A6, CYP2C9, CYP2D6, CYP2E1 or CYP3A4. 2. Ethosuximide was incubated with each expression system at 37 degrees C, and its hydroxylated metabolite was quantified by HPLC assay. 3. K(m) and V(max) values for metabolite formation were estimated for CYP3A4 and CYP2E1. The K(m) values for the metabolite formed were 1.40 and 0.24 mM for CYP3A4 and CYP2E1, respectively. The V(max) values were 0.65 and 0.14 nmol mg-1 protein min(-1) for CYP3A4 and CYP2E1, respectively. 4. These parameters could not be measured for other enzymes, since metabolite concentrations formed were below the HPLC detection limits. 5. Immuno-inhibition studies using specific antibodies against CYP3A4 or CYP2E1 revealed that ethosuximide metabolite levels decreased when the amount of added CYP3A4 or CYP2E1 antibody increased, with anti-CYP3A4 antibodies yielding a greater inhibitory effect. 6. Simulations of scaled-up in vivo ethosuximide CL(hepatic) mediated by CYP3A4 and CYP2E1 based on the in vitro CL'(int) values, which were calculated from the foregoing respective V(max) and K(m) values, project that well over 90% of in vivo CL(hepatic) is due to CYP3A4. These results support an important role for CYP3A in human ethosuximide metabolism, and a minor role for CYP2E1. PMID- 12637245 TI - Non-synonymous single nucleotide alterations in the microsomal epoxide hydrolase gene and their functional effects. AB - 1. By sequencing genomic DNA from 72 established cell lines derived from Japanese individuals, we detected 25 single nucleotide alterations in the microsomal epoxide hydrolase (EPHX1) gene. Of them, five were exonic alterations resulting in amino acid alterations (77C>G, T26S; 128G>C, R43T; 337T>C, Y113H; 416A>G, H139R; 823A>G, T275A). The T26S, R43T, Y113H and H139R substitutions were found at relatively high frequencies and seemed to be polymorphic, and T26S and T275A were novel. 2. To examine the effects of these amino acid alterations on EPHX1 function, EPHX1 cDNA constructs of wild-type and five variants were transfected into COS-1 cells, and their hydrolytic activities for cis-stilbene oxide were determined in vitro. Although all of the transfectants expressed EPHX1 mRNA and protein at similar levels, the variant H139R protein was expressed at a significantly higher level (128% of the wild-type). K(m) values were not significantly different between the wild-type and variants. 3. Increase (140%) in the enzymatic activity (V(max)) of the variant H139R was accompanied by the increased EPHX1 protein level without any significant change in the intrinsic EPHX1 activity. On the other hand, the variant R43T showed increased values for V(max) and clearance (V(max)/K(m)) (around 130%) both on a microsomal protein basis and on a EPHX1 protein basis. 4. These results suggest that R43T as well as H139R increase epoxide hydrolase activity. PMID- 12637246 TI - Metabolism of a novel nucleoside analogue, OGT 719, in the isolated perfused rat liver model, in rats, in tumour models and in patients. AB - 1. The metabolic pathway(s) of OGT 719, a novel nucleoside analogue in which galactose is covalently attached to the N1 of 5-fluorouracil (FU), have been investigated with (19)F-NMR spectroscopy in (1) the isolated perfused rat liver (IPRL) model, (2) normal rats, (3) rats bearing the HSN LV10 sarcoma, (4) nude mice xenografted with the human hepatoma HepG2 and (5) urine from patients. 2. The administration of OGT 719 results in the formation of small amounts of FU. IPRL experiments with OGT 719 in combination with asialofetuin, a natural asialoglycoprotein receptor (ASGP-r), suggest competitive binding of OGT 719 to the ASGP-r. 3. The data obtained in non-tumour rats also demonstrated an extremely low metabolization of OGT 719 into FU and alpha-fluoro-beta-alanine, the well-known major metabolite of FU. 4. A comparison of tumour extracts from rats bearing the HSN LV10 sarcoma treated with FU or OGT 719 showed the incorporation of FU into RNA in rats treated with FU but not in rats treated with OGT 719; nevertheless, the incorporation of FU into RNA was observed in the liver from rats treated with OGT 719. 5. In a human hepatoma xenografted to nude mice, both the OGT 719 and FU contents of the tumour were markedly higher than in the corresponding liver, suggesting a tumour-specific trapping of OGT 719 in hepatoma. 6. The metabolism of OGT 719 was also extremely low in patients. 7. In conclusion, the present study shows the value of (19)F-NMR for demonstrating for the first time that OGT 719 is a prodrug of FU although very poorly metabolized. PMID- 12637247 TI - Single- and repeated-dose pharmacokinetics of eplerenone, a selective aldosterone receptor blocker, in rats. AB - 1. The pharmacokinetics of eplerenone (EP) were examined in rats following single or repeated dosing with (14)C-labelled or unlabelled EP to characterize absorption, metabolism and excretion. Rates of EP metabolism and cytochrome P450 activities were determined in vitro after repeated dose administration of EP. 2. Following a single i.v. dose (15 mg kg(-1)), the elimination half-life of EP was 0.80 and 1.14 h in male and female rats, respectively. Plasma clearances (CL) of EP were 1.62 and 1.20 l kg(-1) h(-1) in males and females, respectively. Following a single oral dose (15 mg kg(-1)), C(max) and T(max) of EP were 1.71 micro g ml(-1) and 0.5 h in male rats. The corresponding values in female rats were 3.54 micro g ml(-1) and 1.0 h. The systemic availability of EP was 25.6% in male rats and 66.4% in female rats, demonstrating sex differences in the pharmacokinetics of EP. 3. In the 8-day study, the AUC(0-24h)'s of total EP (closed lactone ring form plus open form) following 100 and 200 mg kg(-1) oral doses were approximately half those on day 1 in male rats. After repeated dosing for 13 weeks, the pharmacokinetics of total EP did not change with study duration at the 20 mg kg(-1) dose in both males and females. However, at the 100 mg kg(-1) dose, AUC(0-24h)'s were notably reduced on day 24 but progressively increased on subsequent days to approximate day 1 levels by day 86 in both sexes. At the 500 mg kg(-1) dose, the AUCs on day 86 remained lower than those on day 1. Reductions in AUCs on days 8 and 24 appeared to be the result of metabolism induction. 4. EP was extensively metabolized in male rats and most faecal and urinary radioactivity was in the form of metabolites. In female rats, the vast majority of urine and faecal radioactivity was associated with total EP. Thus, the sex difference in the pharmakokinetics of EP was due to more extensive metabolism in male rats. 5. The major metabolite in the rat was 6beta-OH EP. EP 6beta hydroxylase activity was well correlated with testosterone 6beta-hydroxylase activity, indicating that EP metabolism to 6beta-OH EP was mediated primarily by CYP3A in the rat. 6. After repeated dose administration, EP increased 6beta hydroxylase activities of testosterone and EP itself in a dose-dependent manner in both male and female rats, indicating that EP was a CYP3A inducer in the rat. There appeared to be no effects on activities of CYP1A1, 2B and 2E1. PMID- 12637248 TI - Disposition of a synthetic analogue of lipid A (E5564) in rats. AB - 1. E5564, a lipid A analogue that potently antagonises lipopolysaccharide, is being developed to treat sepsis caused by Gram-negative bacterial infections. The pharmacokinetic profile of E5564 is independent of dose between 0.1 and 1 mg kg( 1). The distribution volume of E5564 is slightly larger than the total plasma volume, and the terminal elimination half-life is about 5 h. 2. Following (14)C E5564 administration (0.5 mg kg(-1)), radioactivity rapidly accumulates in the liver and spleen. The half-life of E5564 in the liver is 5.1 h, which is similar to that in the plasma. At 48 weeks after dosing, 35.27% of the administered radioactivity was still present in the liver. Cumulative urinary and faecal excretion of radioactivity for up to 48 weeks after administration were 3.86 and 67.17% of the dose, respectively. 3. The results of mass spectroscopy and nuclear magnetic resonance analysis reveal that the main hepatic metabolite is di dephosphorylated E5564. The half-life of di-dephosphorylated E5564 in the liver is 87.4 days, which is similar to that for the hepatic radioactivity. 4. The results indicate that E5564 is rapidly taken up by the liver, is metabolized via dephosphorylation pathways to form dephosphorylated E5564 and is mainly excreted in the faeces. PMID- 12637249 TI - Role for complement in mediating intestinal nitric oxide synthase-2 and superoxide dismutase expression. AB - Inducible nitric oxide synthase (iNOS) and superoxide dismutase (SOD) play an important role in the pathology of ischemia-reperfusion. This study sought to determine if the proinflammatory effects of complement modulate iNOS and SOD in the rat after gastrointestinal ischemia and reperfusion (GI/R). An inhibitory or noninhibitory anti-complement component 5 (C5) monoclonal antibody (18A or 16C, respectively) was administered before GI/R. RT-PCR revealed a significant increase in intestinal iNOS mRNA compared with sham after GI/R that was attenuated significantly by 18A. Immunohistochemistry demonstrated increased iNOS protein expression within the intestinal crypts after GI/R. Cu/Zn SOD (mRNA and protein) was unaffected by GI/R, whereas Cu/Zn SOD activity was reduced significantly. Mn SOD protein expression was decreased significantly by GI/R. Anti-C5 preserved Cu/Zn SOD activity and Mn SOD protein expression. Staining for nitrotyrosine showed that anti-C5 treatment reduced protein nitration in the reperfused intestine. Immunohistochemistry demonstrated prominent phosphorylated (p) inhibitory factor-kappaB (IkappaB)-alpha staining of intestinal tissue after GI/R, whereas anti-C5 reduced p-IkappaB-alpha expression. These data indicate that complement may mediate tissue damage during GI/R by increasing intestinal iNOS and decreasing the activity and protein levels of Cu/Zn SOD and Mn SOD, respectively. PMID- 12637250 TI - Amelioration of dextran sulfate colitis by butyrate: role of heat shock protein 70 and NF-kappaB. AB - Butyrate enemas have been demonstrated to ameliorate inflammation in ulcerative colitis. The mechanism of this protective effect of butyrate is not known, and this study examines the effect of butyrate on epithelial function, inducible heat shock protein 70 (HSP70) expression, and NF-kappaB activation in experimental colitis. Colitis was induced in rats by oral dextran sulfate sodium (DSS) and by butyrate or saline enemas. Mucosal barrier function was assessed by electrical resistance and [14C]mannitol permeability. HSP70 production was determined by [35S]methionine labeling, Western blot analysis, and immunohistochemistry. Activation of heat shock factors (HSFs) and NF-kappaB was evaluated by EMSA. Butyrate showed a significant protection against the decrease in cell viability, increase in mucosal permeability, and polymorphonuclear neutrophil infiltration seen in DSS colitis. Butyrate inhibited HSP70 expression in DSS colitis and also inhibited the activation of HSF and NF-kappaB. Thus butyrate enema was found to be cytoprotective in DSS colitis, an effect partly mediated by suppressing activation of HSP70 and NF-kappaB. PMID- 12637251 TI - Glucose alleviates ammonia-induced inhibition of short-chain fatty acid metabolism in rat colonic epithelial cells. AB - Ammonia decreased metabolism by rat colonic epithelial cells of butyrate and acetate to CO2 and ketones but increased oxidation of glucose and glutamine. Ammonia decreased cellular concentrations of oxaloacetate for all substrates evaluated. The extent to which butyrate carbon was oxidized to CO2 after entering the tricarboxylic acid (TCA) cycle was not significantly influenced by ammonia, suggesting there was no major shift toward efflux of carbon from the TCA cycle. Ammonia reduced entry of butyrate carbon into the TCA cycle, and the proportion of CoA esterified with acetate and butyrate correlated positively with the production of CO2 and ketone bodies. Also, ammonia reduced oxidation of propionate but had no effect on oxidation of 3-hydroxybutyrate. Inclusion of glucose, lactate, or glutamine with butyrate and acetate counteracted the ability of ammonia to decrease their oxidation. In rat colonocytes, it appears that ammonia suppresses short-chain fatty acid (SCFA) oxidation by inhibiting a step before or during their activation. This inhibition is alleviated by glucose and other energy-generating compounds. These results suggest that ammonia may only affect SCFA metabolism in vivo when glucose availability is compromised. PMID- 12637252 TI - Intestinal epithelial CD23 mediates enhanced antigen transport in allergy: evidence for novel splice forms. AB - We previously demonstrated enhanced transepithelial antigen transport in the intestine of allergic rodents associated with elevated expression of the low affinity IgE receptor CD23 on enterocytes. Here, we examined the role of CD23 in the transport phenomenon using CD23-/- mice and characterized the isoform of intestinal epithelial CD23. Jejunal segments of sensitized mice were challenged with antigen. Enhanced transepithelial antigen transport and transmucosal antigen flux were found in the intestine of sensitized CD23+/+ but not CD23-/- mice. RT PCR showed that enterocytes expressed only the isoform b of CD23. Sequencing revealed classic and alternative CD23b transcripts lacking exon 5 (bDelta5) or 6, all of which were translated into functional IgE receptors. The protein encoded by bDelta5 but not the classic b transcript was able to mediate the uptake of anti-CD23 or IgE, whereas both CD23 proteins were internalized after binding to IgE/antigen complexes. Our results suggest that the classic and alternative forms of CD23b display distinct endocytic properties, suggesting that they are likely to play different roles in transepithelial transport of IgE and allergens. PMID- 12637253 TI - Curcumin attenuates DNB-induced murine colitis. AB - Numerous therapies used for inflammatory bowel disease (IBD) target the transcription factor NF-kappaB, which is involved in the production of cytokines and chemokines integral for inflammation. Here we show that curcumin, a component of the spice turmeric, is able to attenuate colitis in the dinitrobenzene sulfonic acid (DNB)-induced murine model of colitis. When given before the induction of colitis it reduced macroscopic damage scores and NF-kappaB activation. This was accompanied by a reduction in myeloperoxidase activity, and using semiquantitative RT-PCR, an attenuation of the DNB-induced message for IL 1beta was detected. Western blotting analysis revealed that there was a reproducible DNB-induced activation of p38 MAPK detected in intestinal lysates by using a phosphospecific antibody. This signal was significantly attenuated by curcumin. Furthermore, we show that the immunohistochemical signal is dramatically attenuated at the level of the mucosa by curcumin. We conclude that the widely used food additive curcumin is able to attenuate experimental colitis through a mechanism correlated with the inhibition of the activation of NF-kappaB and effects a reduction in the activity of p38 MAPK. We propose that this agent may have therapeutic implications for human IBD. PMID- 12637254 TI - Gastritis increases resistance to aspirin-induced mucosal injury via COX-2 mediated lipoxin synthesis. AB - Products of cyclooxygenase (COX)-2 contribute to mucosal defense. Acetylation of COX-2 by aspirin has been shown to result in the generation of 15(R)-epi-lipoxin A4, which exerts protective effects in the stomach. In gastritis, it is possible that lipoxin A4 makes a greater contribution to mucosal defense. We tested this hypothesis in the rat, by using the iodoacetamide-induced gastritis model. Iodoacetamide was added to the drinking water for 5 days. Rats were then given aspirin, and the extent of gastric damage was blindly assessed 3 h later. Gastric 15(R)-epi-lipoxin A4 and PGE2 levels were determined. The effects of pretreatment with a selective COX-2 inhibitor, rofecoxib, and of a lipoxin receptor antagonist were assessed. Effects of aspirin and the other test drugs on leukocyte adherence within mesenteric venules were assessed by intravital microscopy. Aspirin elicited greater lipoxin synthesis in the inflamed than in the normal stomach, and there was reduced gastric damage. Rofecoxib inhibited lipoxin synthesis and exacerbated aspirin-induced damage. The lipoxin antagonist also exacerbated aspirin-induced damage. In rats with gastritis, aspirin reduced leukocyte adherence (in contrast to an increase in normal rats), and this effect was reversed by rofecoxib or by the lipoxin antagonist. These results support the notion that aspirin-triggered lipoxin synthesis via COX-2 makes an important contribution to mucosal defense in both the normal and inflamed stomach. PMID- 12637255 TI - Androgen therapy improves muscle mass and strength but not muscle quality: results from two studies. AB - The relationship of strength to muscle area was used to assess change in muscle quality after anabolic interventions. Study 1: asymptomatic human immunodeficiency virus-positive men (39 +/- 9 yr) were randomized to nandrolone (600 mg/wk) +/- resistance training (RT). Study 2: older healthy men (72 +/- 5 yr) were randomized to oxandrolone (20 mg/day) or placebo. Maximum voluntary strength was determined by the 1-repetition maximum (1-RM) method for leg press, flexion and extension, and cross-sectional area of leg muscles by MRI. From study week 0 to study week 12, muscle quality was unchanged with nandrolone, oxandrolone, or oxandrolone placebo, respectively, for total thigh muscles (1.23 +/- 0.012 vs. 1.27 +/- 0.29 kg/cm2; 9.0 +/- 1.1 vs. 8.9 +/- 1.2 N/cm2; 8.9 +/- 1.2 vs. 8.9 +/- 1.9 N/cm2) and hamstrings (0.41 +/- 0.08 vs. 0.43 +/- 0.07 kg/cm2; 0.90 +/- 0.14 vs. 0.95 +/- 0.016 N/cm2; 0.94 +/- 0.23 vs. 0.93 +/- 0.21 N/cm2). Lower-extremity 1-RM strength increased several times greater with RT+nandrolone (51-63% increases) than with nandrolone alone (4.7-16%), despite similar increases in muscle area; therefore, muscle quality increased from 1.13 +/- 0.17 to 1.51 +/- 0.18 kg/cm2 (+36 +/- 19%; P < 0.001) for total thigh muscle, 0.37 +/- 0.10 to 0.53 +/- 0.08 kg/cm2 (+49 +/- 39%; P < 0.001) for hamstrings, and 0.73 +/- 0.19 to 1.07 +/- 0.16 kg/cm2 (+55 +/- 36%; P < 0.001) for quadriceps. Thus androgen therapy alone did not improve muscle quality, but the addition of RT to nandrolone produced substantive improvements. PMID- 12637256 TI - p38 MAP kinase regulates BMP-4-stimulated VEGF synthesis via p70 S6 kinase in osteoblasts. AB - We previously reported that p70 S6 kinase takes part in bone morphogenetic protein-4 (BMP-4)-stimulated vascular endothelial growth factor (VEGF) synthesis in osteoblast-like MC3T3-E1 cells. Recently, we showed that BMP-4-induced osteocalcin synthesis is regulated by p44/p42 MAP kinase and p38 MAP kinase in these cells. In the present study, we investigated whether the MAP kinases are involved in the BMP-4-stimulated synthesis of VEGF in MC3T3-E1 cells. PD-98059 and U-0126, inhibitors of the upstream kinase of p44/p42 MAP kinase, failed to affect BMP-4-stimulated VEGF synthesis. SB-203580 and PD-169316, inhibitors of p38 MAP kinase, significantly reduced VEGF synthesis, whereas SB-202474, a negative control for p38 MAP kinase inhibitor, had little effect on VEGF synthesis. The BMP-4-stimulated phosphorylation of p38 MAP kinase was not affected by rapamycin, an inhibitor of p70 S6 kinase. On the contrary, SB-203580 and PD-169316 reduced the BMP-4-stimulated phosphorylation of p70 S6 kinase. In addition, anisomycin, an activator of p38 MAP kinase, phosphorylates p70 S6 kinase, and the phosphorylation was suppressed by SB-203580. LY-294002, an inhibitor of phosphatidylinositol 3-kinase, failed to suppress the phosphorylation of p38 MAP kinase induced by BMP-4. Not BMP-4 but anisomycin weakly induced the phosphorylation of phosphoinositide-dependent kinase-1. However, anisomycin had little effect on phosphorylation of either Akt or the mammalian target of rapamycin. Taken together, our results suggest that p38 MAP kinase functions in BMP-4-stimulated VEGF synthesis as a positive regulator at a point upstream from p70 S6 kinase in osteoblasts. PMID- 12637258 TI - Glucose-induced suppression of endogenous glucose production: dynamic response to differing glucose profiles. AB - To determine whether, in the presence of constant insulin concentrations, a change in glucose concentrations results in a reciprocal change in endogenous glucose production (EGP), glucagon ( approximately 130 ng/l) and insulin ( approximately 65 pmol/l) were maintained at constant "basal" concentrations while glucose was clamped at approximately 5.3 mM (euglycemia), approximately 7.0 mM (sustained hyperglycemia; n = 10), or varied to create a "postprandial" profile (profile; n = 11). EGP fell slowly over the 6 h of the euglycemia study. In contrast, an increase in glucose to 7.13 +/- 0.3 mmol/l resulted in prompt and sustained suppression of EGP to 9.65 +/- 1.21 micromol x kg-1 x min-1. On the profile study day, glucose increased to a peak of 11.2 +/- 0.5 mmol/l, and EGP decreased to a nadir of 6.79 +/- 2.54 micromol x kg-1 x min-1 by 60 min. Thereafter, the fall in glucose was accompanied by a reciprocal rise in EGP to rates that did not differ from those observed on the euglycemic study day (11.31 +/- 2.45 vs. 12.11 +/- 3.21 micromol x kg-1 x min-1). Although the pattern of change of glucose differed markedly on the sustained hyperglycemia and profile study days, by design the area above basal did not. This resulted in equivalent suppression of EGP below basal (-1,952 +/- 204 vs. -1,922 +/- 246 mmol. kg-1. 6 h 1). These data demonstrate that, in the presence of a constant basal insulin concentration, changes in glucose within the physiological range rapidly and reciprocally regulate EGP. PMID- 12637259 TI - Smoking impairs muscle recovery from exercise. AB - Cigarette smoking is a leading cause of many adverse health consequences. Chronic nicotine exposure leads to insulin resistance and may increase the risk of developing non-insulin-dependent diabetes mellitus in young otherwise healthy smokers. To evaluate smoking-induced effects on carbohydrate metabolism, we studied muscle glycogen recovery from exercise in a young healthy population of smokers. The study used 31P-13C NMR spectroscopy to compare muscle glycogen and glucose 6-phosphate levels during recovery in exercised gastrocnemius muscles of randomized cohorts of healthy male smokers (S) and controls (C). Data for the two groups were as follows: S, > or =20 cigarettes/day (n = 8), 24 +/- 2 yr, 173 +/- 3 cm, 70 +/- 4 kg and age- and weight-matched nonsmoking C (n = 10), 23 +/- 1 yr, 175 +/- 3 cm, 67 +/- 3 kg. Subjects performed single-leg toe raises to deplete glycogen to approximately 20 mmol/l, and glycogen resynthesis was measured during the first 4 h of recovery. Plasma samples were assayed for glucose and insulin at rest and during recovery. Test subjects were recruited from the general community surrounding Yale University. Glycogen was depleted to similar levels in the two groups [23.5 +/- 1.2 (S) and 19.1 +/- 1.3 (C) mmol/l]. During the 1st h of recovery, glycogen synthesis rates were similar [13.8 +/- 1.1 (S) and 15.3 +/- 1.3 (C) mmol x l-1 x h-1]. Between hours 1 and 4, glycogen synthesis was impaired in smokers [0.8 +/- 0.2 (S) and 4.5 +/- 0.5 (C) mmol x l-1 x h-1, P = 0.0002] compared with controls. Glucose 6-phosphate was reduced in smokers during hours 1 4 [0.105 +/- 0.006 (S) and 0.217 +/- 0.019 (C) mmol/l, P = 0.0212]. We conclude that cigarette smoking impairs the insulin-dependent portion of muscle recovery from glycogen-depleting exercise. This impairment likely results from a reduction in glucose uptake. PMID- 12637257 TI - Impaired oxidative phosphorylation in skeletal muscle of intrauterine growth retarded rats. AB - Intrauterine growth retardation (IUGR) has been linked to the development of type 2 diabetes in later life. We have developed a model of uteroplacental insufficiency, a common cause of intrauterine growth retardation, in the rat. Early in life, the animals are insulin resistant and by 6 mo of age they develop diabetes. Glycogen content and insulin-stimulated 2-deoxyglucose uptake were significantly decreased in muscle from IUGR rats. IUGR muscle mitochondria exhibited significantly decreased rates of state 3 oxygen consumption with pyruvate, glutamate, alpha-ketoglutarate, and succinate. Decreased pyruvate oxidation in IUGR mitochondria was associated with decreased ATP production, decreased pyruvate dehydrogenase activity, and increased expression of pyruvate dehydrogenase kinase 4. Such a defect in IUGR mitochondria leads to a chronic reduction in the supply of ATP available from oxidative phosphorylation. Impaired ATP synthesis in muscle compromises energy-dependent GLUT4 recruitment to the cell surface, glucose transport, and glycogen synthesis, which contribute to insulin resistance and hyperglycemia of type 2 diabetes. PMID- 12637260 TI - Regulation of translation initiation by insulin and amino acids in skeletal muscle of neonatal pigs. AB - Previous studies have shown that intravenous infusion of insulin and/or amino acids reproduces the feeding-induced stimulation of muscle protein synthesis in neonates and that insulin and amino acids act independently to produce this effect. The goal of the present study was to delineate the regulatory roles of insulin and amino acids on muscle protein synthesis in neonates by examining translational control mechanisms, specifically the eukaryotic translation initiation factors (eIFs), which enable coupling of initiator methionyl-tRNAi and mRNA to the 40S ribosomal subunit. Insulin secretion was blocked by somatostatin in fasted 7-day-old pigs (n = 8-12/group), insulin was infused to achieve plasma levels of approximately 0, 2, 6, and 30 microU/ml, and amino acids were clamped at fasting or fed levels or, at the high insulin dose, below fasting. Both insulin and amino acids increased the phosphorylation of ribosomal protein S6 kinase (S6K1) and the eIF4E-binding protein (4E-BP1), decreased the binding of 4E BP1 to eIF4E, increased eIF4E binding to eIF4G, and increased fractional protein synthesis rates but did not affect eIF2B activity. In the absence of insulin, amino acids had no effect on these translation initiation factors but increased the protein synthesis rates. Raising insulin from below fasting to fasting levels generally did not alter translation initiation factor activity but raised protein synthesis rates. The phosphorylation of S6K1 and 4E-BP1 and the amount of 4E-BP1 bound to eIF4E and eIF4E bound to eIF4G were correlated with insulin level, amino acid level, and protein synthesis rate. Thus insulin and amino acids regulate muscle protein synthesis in skeletal muscle of neonates by modulating the availability of eIF4E for 48S ribosomal complex assembly, although other processes also must be involved. PMID- 12637261 TI - Acute inflammation increases selective uptake of HDL cholesteryl esters into adrenals of mice overexpressing human sPLA2. AB - The acute-phase protein secretory phospholipase A2 (sPLA2) influences the metabolism of high-density lipoproteins (HDL). The adrenals are known to utilize HDL cholesterol as a source of sterols. The aim of the present study was to test the hypothesis that sPLA2 enhances the selective uptake of HDL into the adrenals in response to acute inflammation as a possible physiological role for the sPLA2 HDL interaction. Human sPLA2-transgenic mice, in which sPLA2 expression is upregulated by inflammatory stimuli, were used. Ten hours after induction of the acute-phase response (APR) by injection of bacterial lipopolysaccharide (LPS), plasma levels of HDL cholesterol decreased significantly in sPLA2-transgenic mice (-18%, P < 0.05) but remained unchanged in wild-type mice. The fractional catabolic rates of both 125I-labeled tyraminecellobiose (TC)-HDL and [3H]cholesteryl ether increased significantly in the sPLA2-transgenic mice after induction of the APR (0.18 +/- 0.01 vs. 0.21 +/- 0.01 pool/h, P < 0.05, and 0.31 +/- 0.02 vs. 0.42 +/- 0.05 pool/h, P < 0.05, respectively) but remained unchanged in the wild-type mice (0.10 +/- 0.01 vs. 0.22 +/- 0.02 pool/h, respectively). After induction of the APR, in both groups HDL holoparticle uptake by the liver was increased (P < 0.001). sPLA2-transgenic mice had 2.4-fold higher selective uptake into the adrenals after induction of the APR than wild-type mice (156 +/- 6 vs. 65 +/- 5%/ micro g tissue protein, P < 0.001). In summary, upregulation of sPLA2 expression during the APR specifically increases the selective uptake of HDL cholesteryl ester into the adrenals. These data suggest a novel metabolic role for sPLA2: modification of HDL during the APR to promote increased adrenal uptake of HDL cholesteryl ester to serve as source for steroid hormone synthesis. PMID- 12637262 TI - Influence of K-Cl cotransporter activity on activation of volume-sensitive Cl- channels in human osteoblasts. AB - The whole cell recording mode of the patch-clamp technique was used to study the effect of hypotonic NaCl or isotonic high-KCl solution on membrane currents in a human osteoblast-like cell line, C1. Both hypotonic NaCl or isotonic high-KCl solution activated Cl- channels expressed in these cells as described previously. The reversal potential of the induced Cl- current is more negative when activated through hypotonic NaCl solution (-47 +/- 5 mV; n = 6) compared with activation through isotonic high-KCl solution (-35 +/- 3 mV; n = 8). This difference can be explained by an increase in intracellular [Cl-] through the activity of a K-Cl cotransporter. Potassium aspartate was unable to activate the current, and furosemide or DIOA suppressed the increase in Cl- current induced by isotonic high-KCl solution. In addition, we used the polymerase chain reaction to demonstrate the presence of KCC1-KCC4 mRNA in the osteoblast-like cell line. From these results, we conclude that human osteoblasts express functional K-Cl cotransporters in their cell membrane that seem to be able to induce the indirect activation of volume-sensitive Cl- channels by KCl through an increase in the intracellular ion concentration followed by water influx and cell swelling. PMID- 12637263 TI - SH oxidation coordinates subunits of rat brain ryanodine receptor channels activated by calcium and ATP. AB - We have reported that ryanodine receptor (RyR) channels display three different responses to cytoplasmic free Ca2+ concentration ([Ca2+]) depending on their redox state (Marengo JJ, Hidalgo C, and Bull R. Biophys J 74: 1263-1277, 1998), with low, moderate, and high maximal fractional open times (Po). Activation by ATP of single RyR channels from rat brain cortex was tested in planar lipid bilayers with 10 or 0.1 microM cytoplasmic [Ca2+]. At 10 microM [Ca2+], low-Po channels presented lower apparent affinity to activation by ATP [[ATP] for half maximal activation (KaATP) = 422 microM] than moderate-Po channels (KaATP = 82 microM). Oxidation of low-Po channels with thimerosal or 2,2'-dithiodipyridine (DTDP) gave rise to moderate-Po channels and decreased KaATP from 422 to 82 microM. At 0.1 microM cytoplasmic [Ca2+], ATP induced an almost negligible activation of low-Po channels. After oxidation to high-Po behavior, activation by ATP was markedly increased. Noise analysis of single-channel fluctuations of low Po channels at 10 microM [Ca2+] plus ATP revealed the presence of subconductance states, suggesting a conduction mechanism that involves four independent subchannels. On oxidation the subchannels opened and closed in a concerted mode. PMID- 12637264 TI - High-temporal-resolution analysis demonstrates that ICAM-1 stabilizes WEHI 274.1 monocytic cell rolling on endothelium. AB - Leukocyte rolling, adhesion, and migration on vascular endothelium involve several sets of adhesion molecules that interact simultaneously. Each of these receptor-ligand pairs may play multiple roles. We examined the role of ICAM-1 in adhesive interactions with mouse aortic endothelial cells (MAECs) in an in vitro flow system. Average rolling velocity of the monocytic cell line WEHI 274.1 was increased on ICAM-1-deficient MAECs compared with wild-type MAECs, both with and without TNF-alpha stimulation. High-temporal-resolution analysis provided insights into the underlying basis for these differences. Without TNF-alpha stimulation, average rolling velocity was slower on wild-type than on ICAM-1 deficient endothelium because of brief (<1 s) pauses. On TNF-alpha-stimulated ICAM-1-deficient endothelium, cells rolled faster because of transient accelerations, producing "jerky" rolling. Firm adhesion to ICAM-1-deficient MAECs was significantly reduced compared with wild-type MAECs, although the number of rolling cells was similar. These results demonstrate directly that ICAM-1 affects rolling velocity by stabilizing leukocyte rolling. PMID- 12637265 TI - Increased susceptibility of cholangiocytes to tumor necrosis factor-alpha cytotoxicity after bile duct ligation. AB - Tumor necrosis factor (TNF)-alpha plays a critical role in epithelial cell injury. However, the role of TNF-alpha in mediating cholangiocyte injury under physiological or pathophysiological conditions is unknown. Thus we assessed the effects of TNF-alpha alone or following sensitization by actinomycin D on cell apoptosis, proliferation, and basal and secretin-stimulated ductal secretion in cholangiocytes from normal or bile duct-ligated (BDL) rats. Cholangiocytes from normal or BDL rats were highly resistant to TNF-alpha alone. However, presensitization by actinomycin D increased apoptosis in cholangiocytes following BDL and was associated with an inhibition of proliferation and secretin stimulated ductal secretion. Thus TNF-alpha mediates cholangiocyte injury and altered ductal secretion following bile duct ligation. These observations suggest that cholestasis may enhance susceptibility to cytokine-mediated cholangiocyte injury. PMID- 12637266 TI - Microtubule-dependent PKC-alpha localization in A7r5 smooth muscle cells. AB - Using laser scanning confocal, fluorescence resonance energy transfer (FRET), and atomic force (AFM) microscopy, we investigated association of protein kinase C (PKC)-alpha with microtubules during stimulus-induced relocalization in A7r5 smooth muscle cells. Confocal microscopy with standard immunostaining techniques confirmed earlier observations that colchicine disruption of microtubules blocked PKC-alpha localization in the perinuclear region of the cell caused by phorbol 12,13-dibutyrate (PDBu; 10-6M). Dual immunostaining suggested colocalization of PKC-alpha and beta-tubulin in both unstimulated and PDBu-treated cells. This finding was verified by FRET microscopy, which indicated that association of PKC alpha was heterogeneous in distribution and confined primarily to microtubules in the perinuclear region. FRET analysis further showed that association between the molecules was not lost during colchicine-induced dissolution of microtubules, suggesting formation of tubulin-PKC-alpha complexes in the cytosol. Confocal imaging indicated that perinuclear microtubular structure was more highly sensitive to colchicine dissolution than other regions of the cell. Topographic imaging of fixed cells by AFM indicated a well-defined elevated structure surrounding the nucleus that was absent in colchicine-treated cells. It was calculated that the volume of the nuclear sleevelike structure of microtubules increased approximately fivefold in PDBu-treated cells, suggesting a probable increase in microtubular mass. In light of PKC-alpha localization, increased colchicine sensitivity, and their volume change in stimulated cells, the results suggest that perinuclear microtubules form a specialized structure that may be more dynamically robust than in other regions of the cell. PKC-alpha could contribute to this dynamic activity. Alternatively, perinuclear microtubules could act as a scaffold for regulatory molecule interaction at the cell center. PMID- 12637267 TI - Molecular cloning and characterization of a rat sensory nerve Ca2+-sensing receptor. AB - A full-length cDNA encoding a Ca2+-sensing receptor (CaSR) expressed in rat dorsal root ganglia (DRG) was identified using rapid amplification of 5'-cDNA ends and primer extension and then cloned into the plasmid vector pCR3.1. The DNA sequence of the DRG CaSR was 99.9% homologous with published rat kidney CaSR in the coding region and 247 bp upstream of the start site but showed little homology 5' to this site, which maps to exonic junction I/II, supporting the hypothesis that CaSR message arises as a splice variant and showing tissue-to tissue heterogeneity. Western blot revealed a doublet of 140 and 160 kDa in a thyroparathyroid preparation and a single 140-kDa band in DRG. Deglycosylation using N-glycanase increased the mobility of CaSR protein from both DRG and thyroparathyroid, whereas endo-H was without effect, indicating that the DGR CaSR is a mature form of the receptor. A DRG CaSR-pEGFP fusion product was constructed, and when transfected into HEK-293 cells, it was distributed at the cell membrane and resulted in extracellular Ca2+ (0.5-3 mM)-evoked increases in intracellular Ca2+, which in some instances exhibited oscillatory behavior. We conclude that DRG CaSR cDNA arises from tissue-specific alternative splicing of a single gene, that the amino acid sequence of DRG CaSR is homologous to other known CaSRs, and that the DRG CaSR undergoes differential posttranslational processing relative to the thyroparathyroid CaSR and is functionally active when transfected into a human-derived cell line. PMID- 12637268 TI - Posttranslational inactivation of human xanthine oxidoreductase by oxygen under standard cell culture conditions. AB - Xanthine oxidoreductase (XOR) catalyzes the final reactions of purine catabolism and may account for cell damage by producing reactive oxygen metabolites in cells reoxygenated after hypoxia. We found a three- to eightfold higher XOR activity in cultured human bronchial epithelial cells exposed to hypoxia (0.5-3% O2) compared with cells grown in normoxia (21% O2) but no difference in XOR protein or mRNA. XOR promoter constructs failed to respond to hypoxia. The cellular XOR activity at 3% O2 returned to basal levels when the cells were returned to 21% O2, and hyperoxia (95% O2) abolished enzyme activity with no change in XOR protein. Our data suggest reversible enzyme inactivation by oxygen or its metabolites. NADH was normally oxidized by the oxygen-inactivated enzyme, which rules out damage to the flavin adenine dinucleotide cofactor. Hydrogen peroxide partially inactivated the molybdenum center of XOR, as shown by a parallel decrease in XOR-catalyzed xanthine oxidation and dichlorophenolindophenol reduction. We conclude that the transcription or translation of XOR is not influenced by hypoxia or hyperoxia. Instead, the molybdenum center of XOR is posttranslationally inactivated by oxygen metabolites in "normal" (21% O2) cell culture atmosphere. This inactivation is reversed in hypoxia and accounts for the apparent induction. PMID- 12637269 TI - Building a better mousetrap: the diagnosis of metastatic cervical adenopathy. PMID- 12637270 TI - Myelography: still the gold standard. PMID- 12637271 TI - Luminal and mural imaging of aneurysms. PMID- 12637273 TI - Combination of helical CT and Doppler sonography in the follow-up of patients with clinical N0 stage neck disease and oral cancer. AB - BACKGROUND AND PURPOSE: Correctly diagnosing metastatic nodes is important for the follow-up of patients with clinical N0 stage neck disease and oral cancer. A combination of helical CT and Doppler sonography may facilitate the accurate detection of lymph node metastasis in patients with clinical N0 stage neck disease. METHODS: A combination of contrast-enhanced helical CT and Doppler sonography was performed to monitor the necks of 58 patients with initial clinical N0 stage neck disease. Of these patients, 17 underwent surgery; nodal metastasis in the neck was histopathologically confirmed. A node was diagnosed as metastatic if it fulfilled the CT criteria for metastatic nodes (short-axis diameter equal to or greater than the cutoff points for each level of the neck or central nodal necrosis) and if, additionally, it did not exhibit sonographic features for nonmetastatic nodes (normal hilar echogenicity and hilar flows). The presence of metastasis was confirmed histopathologically. RESULTS: During the follow-up periods, metastatic nodes were histologically confirmed in 17 (29%) patients. Of 30 metastatic nodes from the 17 patients with metastatic nodes, 22 (73%) appeared within the first year and 28 (93%) within the first 2 years; 20 developed from nonmetastatic nodes, and 10 were newly detectable. The combined criteria were effective in revealing 26 (87%) nodes, yielding 87% sensitivity, 100% specificity, and 100% positive and 99% negative predictive values. The independent use of one of these techniques alone resulted in low (67%) or moderate (87%) positive predictive values for sonography and CT, respectively. Seven hundred forty-one (97%) of 761 nodes that were nonmetastatic at initial examination remained nonmetastatic (737 nodes) or had disappeared (four nodes). As a result, a combination of CT and sonography was effective in revealing all 17 cases of metastatic nodes. CONCLUSION: A combination of contrast-enhanced helical CT and Doppler sonography is useful for the follow-up study of clinical N0 stage neck disease. PMID- 12637272 TI - Assessment of metastatic cervical adenopathy using dynamic contrast-enhanced MR imaging. AB - BACKGROUND AND PURPOSE: Morphologic assessment by conventional imaging methods of lymph node metastases in patients with squamous cell carcinoma of the head and neck is, at best, insensitive. Doppler sonography has shown that lymph node metastases exhibit alterations in the number of vessels and blood flow. We assessed the ability of dynamic contrast-enhanced MR imaging to differentiate normal from diseased nodes in this patient population. METHODS: Twenty-one patients with newly diagnosed squamous cell carcinoma and no previous treatment were studied with the use of a head and neck phased array surface coil. Anatomic imaging included high resolution T1-weighted, fat-saturated fast spin-echo T2 weighted, and contrast-enhanced T1-weighted imaging (0.99-1.32 mm(3) voxels). The dynamic contrast-enhanced MR imaging was performed by using a 2D fast spoiled gradient recalled sequence with single dose bolus injection of contrast agent. Calculated values included time to peak, peak enhancement, maximum slope, and washout slope for the enhancement. All patients underwent neck dissection as part of their indicated treatment, and imaging results were correlated with pathologic findings. RESULTS: Dynamic contrast-enhanced MR imaging and pathology comparisons were obtained for 68 nodes. There was significantly longer time to peak (P <.001), lower peak enhancement (P <.05), lower maximum slope (P <.01), and slower washout slope (P <.05) in the tumor-involved nodes compared with the normal nodes. CONCLUSION: Analysis of dynamic contrast-enhanced MR imaging can differentiate normal from diseased lymph nodes in patients with squamous cell carcinoma of the head and neck. PMID- 12637274 TI - Effects of zero-filled interpolation on cervical magnetic resonance angiographic measurement. AB - BACKGROUND AND PURPOSE: The aim of this study was to assess the effect of zero filled interpolation (ZIP) on measurements of the cervical arteries because its benefits on the accuracy and precision of measurements in medium-sized arteries remains unknown. METHODS: Three observers measured 36 computer-simulated vessels (2-6.8 mm) and 130 normal cervical vessels (assessed with two-dimensional time-of flight MR angiography) from 512-, 256-, and 256-ZIP matrix source images. The accuracy and precision of measurement was assessed for each matrix by using the Student t test and F test of variance, respectively. The effect of vessel size and matrix placement on measurement error was determined by means of linear regression and the Student t test, respectively. RESULTS: No significant difference was observed between simulated measurements obtained on the 512 matrix and their true value. The 256 matrix caused overestimation of vessel diameter compared with 512 matrix (mean bias, 0.3 mm for computer-simulated vessels and 0.1 mm for normal vessels). This effect was reduced with ZIP, by a mean of 0.1 mm for both groups (P <.03). Precision was not affected by the matrix size or ZIP, and vessel size and matrix placement did not alter the measurement error. CONCLUSION: Vessel diameter is overestimated on 256-matrix MR angiographic source images. ZIP reduces this overestimation; however, the effect is small and unlikely to be clinically important. PMID- 12637275 TI - Aortic arch origin of the left external carotid artery and type II proatlantal fetal anastomosis. AB - This case report demonstrates a rare vascular pattern of a left external carotid artery (ECA) that arises directly from the aortic arch (AA), along with a type II proatlantal-vertebral artery anastomosis. Our research shows no report of an ECA arising from the AA. This, in association with a type II proatlantal anastomosis, is interesting because of its apparent rarity. PMID- 12637276 TI - Preoperative facial muscle imaging predicts final facial function after facial nerve grafting. AB - BACKGROUND AND PURPOSE: Our goal was to determine whether preoperative MR imaging of facial muscles predicts facial function after facial nerve grafting. METHODS: A retrospective review of all patients undergoing facial nerve grafting between 1997 and 2001 revealed 26 patients. Twelve of the patients had adequate preoperative MR images available for review and had undergone clinical follow-up for at least 12 months. Eight had malignant parotid tumors, and four had benign skull base or parotid tumors. Preoperative facial muscle MR imaging appearance was categorized as symmetrical or asymmetrical. The asymmetrical images were further classified into mild or pronounced asymmetry. Preoperative facial function was classified by using the House-Brackmann scale. Postoperative function was graded with the May scale. RESULTS: Four patients had symmetrical facial muscles shown by preoperative MR imaging, three had mild asymmetry, and five had pronounced asymmetry. No or mild asymmetry had an 86% positive predictive value for good to excellent functional outcome. Eighty percent of patients with pronounced asymmetry experienced poor functional outcomes. Six of eight patients with malignant and perineural tumors at surgery had asymmetrical facial muscles revealed by preoperative MR imaging studies. CONCLUSION: Symmetrical or mildly asymmetrical facial muscles are predictive of good facial function after nerve grafting. Pronounced asymmetry of facial muscles on MR images is predictive of poor facial function after grafting. Asymmetric facial muscles on preoperative MR images are associated with perineural tumor spread in patients with malignant disease. PMID- 12637277 TI - Cervicofacial actinomycosis: CT and MR imaging findings in seven patients. AB - BACKGROUND AND PURPOSE: Cervicofacial actinomycosis is uncommon, but without proper treatment it causes extensive tissue destruction. Early diagnosis is critical but usually difficult with cultures or imaging. Our aim was to identify characteristic imaging features that facilitated diagnosis in seven patients with cervicofacial actinomycosis. METHODS: We retrospectively reviewed the CT and MR findings in seven patients with pathologically proved actinomycosis. Histologic diagnosis was made by means of surgical excision or biopsy in seven patients. Culturing was performed in two patients. Enhanced CT scans (n=7) and MR images (n=2) were evaluated for the location, margin, infiltrative nature, enhancement pattern, and presence of lymphadenopathy. RESULTS: CT and MR images showed either a well-defined (n=2) or ill-defined (n=5) mass. Involved areas included the nasal cavity (n=2), buccal space (n=1), pyriform sinus (n=1), aryepiglottic fold (n=1), oro- and hypopharynx (n=1) and tongue (n=1). Imaging confirmed the infiltrative nature, showing the tendency of the lesion to invade across tissue planes and boundaries (n=6). Moderate homogeneous contrast enhancement was seen on CT scans in six patients with several small low-attenuating foci (n=2). T1- and T2 weighted MR images showed intermediate signal intensity with moderate contrast enhancement (n=2). Reactive lymphadenopathy was associated in three patients. CONCLUSION: Although cervicofacial actinomycosis occurs infrequently, it should be included in the differential diagnosis when images show a soft-tissue mass with inflammatory changes and an infiltrative nature in the cervicofacial area. PMID- 12637279 TI - Solitary fibrous tumor of the hypoglossal nerve. AB - SUMMARY: We report a case of solitary fibrous tumor (SFT) causing isolated hypoglossal nerve palsy. The neuroimaging appearance of the tumor was indistinguishable from that of schwannoma or meningioma. Immunohistochemical tests demonstrated strong reactivity for CD34 but an absence of staining for S100 and epithelial membrane antigen; this profile is indicative of an SFT. SFTs are mesenchymal tumors that can affect the dura-covered segments of cranial nerves. They may be considered in the differential diagnosis of an isolated cranial nerve palsy. PMID- 12637278 TI - MR imaging study of edema-like change along the optic tract in patients with pituitary region tumors. AB - BACKGROUND AND PURPOSE: Edema-like change along the optic tract commonly occurs in association with craniopharyngiomas. The aim of this study was to clarify whether it occurs in association with other common pituitary region tumors and to elucidate its mechanism as seen on MR images. METHODS: Fifty patients with pituitary region tumors that were touching or compressing the optic pathway underwent heavily T2-weighted MR imaging before and after treatment. RESULTS: Edema-like change along the optic tract was visible on the images of four of 25 pituitary adenomas, eight of 11 craniopharyngiomas, one germ cell tumor, and one malignant lymphoma and was not visible on the images of seven meningiomas and five Rathke's cleft cysts. After therapeutic decompression of the optic pathway, the edema-like change disappeared and large Virchow-Robin spaces, present under normal conditions, became visible along the optic tract. Comparison of pre- and post-treatment coronal and axial view MR images revealed that the edema-like change had been located at, along, and/or around the large Virchow-Robin spaces along the optic tract. CONCLUSION: Edema-like change occurs in association with pituitary region tumors other than craniopharyngiomas. It is related with distension of normally present large Virchow-Robin spaces adjacent to the optic tract. Because Virchow-Robin spaces are speculated to be a drainage route of interstitial fluid into the subarachnoid space, their distension may be related to the fluid retention in and along the Virchow-Robin spaces, the outlet of which into the subpial and/or subarachnoid space(s) is blocked by pituitary region tumors. PMID- 12637280 TI - MR imaging appearance of frontal sinus barotrauma. AB - We present the case of a flight passenger who experienced acute and severe headache during landing. MR imaging was performed because the patient had a history of vascular malformation and revealed an incidental venous angioma. A mass lesion in the frontal sinus, consistent with submucosal hematoma secondary to barotrauma, was thought to be the cause of the headache. To our knowledge, this is the first case of sinus barotrauma described in the radiologic literature and the first to describe the associated MR imaging findings. PMID- 12637281 TI - Lumbar root compression in the lateral recess: MR imaging, conventional myelography, and CT myelography comparison with surgical confirmation. AB - BACKGROUND AND PURPOSE: Previous authors have shown that conventional myelography is superior to plain CT in the assessment of root compression in the lateral recess, but this question has never been evaluated with respect to MR imaging of the lumbar level. Our purpose was to assess the accuracy of MR imaging, conventional myelography, and postmyelography CT (CT myelography) of the lumbar level in identifying degenerative lateral recess root compression with surgical confirmation. METHODS: MR imaging, conventional myelography, and CT myelography of the lumbar level were assessed in the imaging of 58 lateral recesses at 38 lumbar levels in 26 patients who underwent surgery for radiculopathy with degenerative lateral recess abnormality. Each lateral recess was graded as normal, small without root compression, small with root compression, or severe root compression. RESULTS: MR imaging underestimated root compression in 28% to 29% of the cases in which root impingement was surgically confirmed. Conventional myelography underestimated root compression in only 5% to 7% of the cases and correctly predicted impingement in 93% to 95%. CT myelography underestimated root compression in 38% of the surgically confirmed cases. CONCLUSION: MR imaging significantly underestimated root compression caused by degenerative changes in the lateral recess. Although MR imaging is a superb study when used in the search for degenerative disk disease and disk protrusion, conventional myelography is a crucial supplemental study that is necessary to confirm degenerative root impingement in the lateral recess as the cause of radiculopathy. PMID- 12637282 TI - Percutaneous intraspinal navigation: feasibility study of a new and minimally invasive approach to the spinal cord and brain in cadavers. AB - We describe a percutaneous approach for cerebral surgical access. After lumbar puncture, the spinal subarachnoid space was traversed by using standard angiographic guidewire techniques until the introducer catheters were in the intracranial space. Under fluoroscopic guidance, the intracranial subarachnoid space was navigated, and the ventricular system entered. Subarachnoid placement was confirmed with contrast-enhanced digital angiography. Placement anterior to the brain stem was confirmed in both cadavers during dissection, and spinal navigation without cord damage from the anterior or posterior approach was confirmed in one. Percutaneous intraspinal navigation is a new route of access for cerebrospinal surgery that has many potential applications. PMID- 12637283 TI - Contrast enhancement patterns of acute spinal epidural hematomas: a report of two cases. AB - The MR finding of an acute spinal epidural hematoma (SEH) can mimic epidural neoplastic or inflammatory lesions, because acute SEH appears hypointense on T1 weighted images and hyperintense on T2-weighted images. We report on two cases of acute SEH with unusual contrast enhancement patterns on MR images. Contrast enhancement can be an MR finding of acute SEH, especially in bleeding diathesis, which indicates an active lesion that needs early diagnosis and management. PMID- 12637284 TI - Pediatric intracranial aneurysm: a diagnostic dilemma solved with contrast enhanced MR imaging. AB - Childhood intracranial aneurysms are exceedingly uncommon. Diagnosis of intracranial aneurysms in infancy may be difficult because of their infrequency and confusing clinical presentation. Findings with routine radiographic methods may be misleading and difficult to interpret. We present a case of the rupture of an anterior communicating artery aneurysm in a 7-month-old child. The rupture had eluded diagnosis until contrast-enhanced MR imaging was performed. PMID- 12637285 TI - MR ventriculography for the study of CSF flow. AB - BACKGROUND AND PURPOSE: Various MR techniques have been used to assess CSF flow and to image the subarachnoid spaces and ventricles. Anecdotal reports describe the use of intrathecal and intraventricular gadolinium-based contrast agents in humans and animals. We sought to determine the clinical usefulness of gadolinium enhanced MR ventriculography for assessing CSF flow in patients with various neurologic conditions. METHODS: Five patients (three female and two male patients aged 6 months to 65 years) were included in the study. After performing sagittal, coronal, and axial T1-weighted MR imaging of the brain, 0.02-0.04 mmol of gadodiamide was injected into the lateral ventricle. Sagittal, coronal, and axial T1-weighted imaging was repeated soon after the injection. We were specifically looking for the site of obstruction to CSF flow in those patients with hydrocephalus, communication between cysts and ventricles, elucidation of suspicious intraventricular lesions, and patency of third ventriculostomies. RESULTS: MR ventriculography showed good delineation of the ventricular system in all patients. In one patient with carcinomatosis and hydrocephalus, a block to contrast material flow was detected at the right foramen of Luschka. In another patient with hydrocephalus, partial block to the flow of contrast material was demonstrated at the right foramen of Monro. In a patient with hydrocephalus and a posterior fossa cyst, flow of contrast material was blocked between the third ventricle and the cyst, with a thin streak of contrast material in the aqueduct. As an assessment of the patency of a third ventriculostomy, MR ventriculography showed flow of contrast material into the suprasellar cisterns from the third ventricle in one patient and absence of flow in another. CONCLUSION: MR ventriculography is a safe technique for assessing CSF flow, with application in determining the site of obstruction in hydrocephalus, in assessing communication between cysts and the ventricle, and in determining the functioning status of endoscopic third ventriculostomies. PMID- 12637286 TI - Cognitive impairment in children with hemoglobin SS sickle cell disease: relationship to MR imaging findings and hematocrit. AB - BACKGROUND AND PURPOSE: Children with hemoglobin SS sickle cell disease are known to suffer cognitive impairment if they have silent infarct, but recent evidence suggests that patients with hemoglobin SS sickle cell disease may be impaired even if they are free of infarction. We test a hypothesis that cognitive impairment in children with hemoglobin SS sickle cell disease is associated with low hematocrit and MR imaging abnormalities. METHODS: A cohort of 49 patients was examined, all of whom had hemoglobin SS sickle cell disease but no history of clinical stroke. The Wechsler scales, which are standardized and age-adjusted, were used to assess cognitive function. Patients also underwent MR imaging examination of the brain, and hematocrit was measured in a subset of 45 patients. MR images were evaluated by at least two readers, and abnormal imaging findings were evaluated by at least three readers. Any lesion was sufficient to be classified as abnormal, with lesions defined to include lacunar infarction, encephalomalacia, or leukoencephalopathy. Hematocrit data were used if obtained within 3 months of psychometric testing and if there were no confounding events in the patients' charts. Wechsler test scores were then evaluated in relation to imaging findings and hematocrit values. RESULTS: Patients with imaging abnormalities had more cognitive impairment than did patients with normal imaging findings in verbal intelligence quotient (P <.02) and verbal comprehension (P <.01). Patients with low hematocrit had cognitive impairment shown by many performance measures, including full-scale intelligence quotient (P <.006), verbal comprehension (P <.006), and freedom from distractibility (P <.02). Multivariate analysis showed that MR imaging and hematocrit were independent predictors of full-scale intelligence quotient. CONCLUSION: Focal brain injury, revealed by MR imaging, is associated with cognitive impairment, but our data suggest that diffuse brain injury may also contribute to impairment. These findings show that impairment is multifactorial and suggest that chronic brain hypoxia is part of the pathophysiology of sickle cell disease. PMID- 12637287 TI - Brain MR imaging in acute hyperammonemic encephalopathy arising from late-onset ornithine transcarbamylase deficiency. AB - The brain MR imaging characteristics of three patients with acute hyperammonemic encephalopathy resulting from late-onset ornithine transcarbomylase deficiency are presented. MR images revealed injury to the cortex, especially the cingulate gyrus and insular cortex, with sparing of the perirolandic and occipital cortices. These findings presumably reflect the distribution of brain injury from hypoperfusion secondary to hyperammonemia. Knowledge of the MR findings may help expedite diagnosis and treatment and prevent chronic impairment. PMID- 12637288 TI - Middle interhemispheric variant of holoprosencephaly associated with diffuse polymicrogyria. AB - An 11-month-old boy was discovered to have a cleft palate, club foot, hypospadias, and myoclonic seizures. No in utero exposure to teratogens was identified. Brain MR imaging revealed a middle interhemispheric fusion variant of holoprosencephaly, diffuse polymicrogyria, and a hypoplastic brain stem; this was a distinctly unusual association of findings. We hypothesize that an unknown genetic factor causes disturbances of cleavage of the prosencephalon as well as neuronal migration and organization. PMID- 12637289 TI - Infantile sialic acid storage disease: serial ultrasound and magnetic resonance imaging features. PMID- 12637290 TI - Practical visualization of internal structure of white matter for image interpretation: staining a spin-echo T2-weighted image with three echo-planar diffusion-weighted images. AB - BACKGROUND AND PURPOSE: To our knowledge, no method satisfactory for clinical use has been developed to visualize white matter fiber tracts with diffusion-weighted MR imaging. The purpose of this study was to determine whether superposition of a spin-echo T2-weighted image and a color-coded image derived from three orthogonal diffusion-weighted images could show fiber tract architecture of the brain with an image quality appropriate for accurate reading with a computer monitor. METHODS: MR images from 50 consecutive cases were reviewed. Three diffusion weighted images per section were acquired with three orthogonal motion-probing gradients. These images were registered to a corresponding spin-echo T2-weighted image. A color-coded image was synthesized from three diffusion-weighted images by assigning red, green, or blue to each diffusion-weighted image and then adding a spin-echo T2-weighted image with a weighting factor. The ability of the superposed image to delineate the white matter pathways was evaluated on the basis of the known anatomy of these pathways and qualitatively compared with that of the spin-echo T2-weighted image. RESULTS: The main white matter fiber pathways, in particular the superior longitudinal fascicle, corpus callosum, tapetum, optic radiation, and internal capsule, were more clearly and easily identified on the superposed image than on the spin-echo T2-weighted image. The time required to produce the superposed image was approximately 40 minutes. CONCLUSION: Superposition of a spin-echo T2-weighted image and a color-coded image created from three orthogonal diffusion-weighted images showed structures of the brain that were not clearly visible on the spin-echo T2-weighted image alone. Such superposition presents images that are easy to interpret correctly. PMID- 12637291 TI - Reliable callosal measurement: population normative data confirm sex-related differences. AB - BACKGROUND AND PURPOSE: Corpus callosal cross-sectional area (CCA) may be a clinical indicator of disease progression, but factors influencing callosal morphology in healthy subjects must be determined before comparisons can be made in patients. We sought to define a reliable and easily repeatable method for CCA measurement and to examine the effects of sex, age, handedness, and cerebral volume. METHODS: Neurologically healthy volunteers (age range, 14-68 years; mean age, 32.6 years +/- 12.3 [SD]; 44 men, 56 women; 87 right handed) underwent conventional MR imaging. Data were reoriented in the image space to account for intersubject variations in head position before the midsagittal plane was defined by using midpoints of the anterior commissure (AC), posterior commissure (PC), and interhemispheric fissure (IF). Midsagittal CCA and total cerebral volume were measured and correlated with sex, age, and handedness. RESULTS: The mean CCA was 6.27 cm(2) +/- 0.90. Women had a larger CCA proportional to cerebral volume (6.16 x 10(-3) cm(-1) vs 5.78 x 10(-3) cm(-1) in men; P =.02). The percentage difference for the CCA-cerebral volume from the group mean was +2.6% in women and -3.6% in men. Only a small linear relationship of CCA with cerebral volume was noted (r(2) = 0.15), and CCA was not significantly correlated with age or handedness. CONCLUSION: To our knowledge, this is the largest study of callosal area in a community-based sample of control subjects; such subjects provide controls for future studies. Our findings provide anatomic evidence of sex differences in interhemispheric connectivity. Much CCA variability is independent of cerebral volume. PMID- 12637292 TI - Quantitative cerebral blood flow measurement with dynamic perfusion CT using the vascular-pixel elimination method: comparison with H2(15)O positron emission tomography. AB - BACKGROUND AND PURPOSE: Blood vessels are usually conspicuous on dynamic CT perfusion images. The presence of large vessels may lead to overestimation of the quantitative value of cerebral blood flow (CBF). We evaluated the efficacy of the vascular-pixel elimination (VPE) method in quantitative CT perfusion imaging, in comparison with positron emission tomography (PET). METHODS: Five healthy volunteers underwent CT perfusion and PET studies. A four-channel multi-detector row CT scanner was used. Dynamic cine scanning was performed after bolus injection of an intravenous contrast agent. CT-CBF was calculated by the central volume principle and deconvolution method. PET was performed after infusion of (15)O-labeled water. PET-CBF was calculated by using a nonlinear least squares method. Average CBF values of the whole section, gray matter, and white matter with both CT and PET were compared after image registration. The comparison was performed with and without VPE. In the VPE method, the vascular pixels were defined by the cerebral blood volume value of the pixel. The threshold of VPE was changed from 5 to 20 mL/100 g. Pixel-by-pixel correlation between CT-CBF and PET CBF and linear regression analysis were also performed. RESULTS: Without VPE, CT CBF was overestimated in all subjects. As the VPE threshold decreased, CT-CBF decreased and the correlation coefficient increased. The best correlation was observed at a VPE threshold of 8 mL/100 g in four of the five subjects. Average CT-CBF values, without VPE, of the whole section, gray matter, and white matter were 59.01, 66.73, and 42.53 mL/100 g/min, respectively. With VPE (threshold, 8 mL/100 g), average CT-CBF values of the whole section, gray matter, and white matter were 45.56, 52.75, and 30.38, respectively. The corresponding PET-CBF values were 46.86, 50.89, and 38.20 mL/100 g/min, respectively. CONCLUSION: Vascular pixels should be excluded from the calculation of CT-CBF to avoid overestimation of the CBF values. If vascular pixels are excluded, CBF calculation with CT perfusion imaging is considerably accurate. PMID- 12637293 TI - Topographic distribution of misery perfusion in relation to internal and superficial borderzones. AB - BACKGROUND AND PURPOSE: Whether misery perfusion (MP) commonly accompanies brain borderzones (BZs) in patients with major cerebral artery occlusion remains unclear. We elucidated topographic patterns of chronic hemodynamic failure in such patients. METHODS: Twenty-four patients with unilateral occlusion or severe stenosis (>75% in diameter) of the internal carotid artery (ICA) or middle cerebral arterial (MCA) trunk with minimal or no infarct underwent PET with (15)O labeled gas inhalation. Mean cerebral blood flow (CBF), cerebral blood volume (CBV), cerebral metabolic rate of oxygen, oxygen extraction fraction (OEF), and CBV/CBF ratio were determined in the superficial BZs, internal BZ, and MCA territory excluding BZs. Values in BZs were standardized and compared with those in controls. Topographic distributions of regions with OEF greater than that in controls were determined. RESULTS: Values in patients and controls were not significantly different. Topographic distributions included matched perfusion in 10 patients, MP in only the ipsilateral internal BZ in four, MP in both ipsilateral internal and superficial BZs in two, MP in the ipsilateral MCA territory including BZs in one, MP in the ipsilateral MCA territory including BZs and contralateral BZs in two, and MP in the ipsilateral MCA territories including BZs in five. CONCLUSION: Only 25% of the patients had MP localized in affected BZs Although localized MP more frequently accompanied the internal BZ than other regions, no patient had elevated OEF in the superficial BZ alone. These results are inconsistent with clinical observations that 80% of BZ infarctions develop superficially. Thus, hemodynamic mechanisms may not cause most superficial BZ infarctions. PMID- 12637294 TI - Assessing tissue viability with MR diffusion and perfusion imaging. AB - BACKGROUND AND PURPOSE: Diffusion- (DW) and perfusion-weighted (PW) MR imaging reflect neurophysiologic changes during stroke evolution. We sought to determine parameters that distinguish regions of brain destined for infarction from those that will survive despite hypoperfusion. METHODS: DW and PW images were obtained in 30 patients at 1-12 hours after symptom onset. Relative cerebral blood volume (rCBV), flow (rCBF), mean transit time (MTT), apparent diffusion coefficient (ADC), DW image signal intensity, and fractional anisotropy (FA) lesion contralateral normal region ratios were obtained in the following regions: 1) infarct core with hyperintensity on DW image, abnormality on rCBF and MTT images, and follow-up abnormality; 2) infarcted penumbra with normal DW image, abnormal rCBF and MTT images, and follow-up abnormality; and 3) hypoperfused tissue that remained viable, with normal DW image, abnormal rCBF and MTT images, and normal follow-up. RESULTS: rCBF ratios for regions 1, 2, and 3 were 0.32 +/- 0.11, 0.46 +/- 0.13, and 0.58 +/- 0.12, respectively, and were significantly different. DW image intensity and ADC ratios were significantly different among all regions, but were more similar than rCBF ratios. rCBV and FA ratios were not significantly different between regions 2 and 3. No MTT ratios were significantly different. No region of interest with an rCBF ratio less than 0.36, an rCBV ratio less than 0.53, an ADC ratio less than 0.85, a DW image intensity ratio greater than 1.23, or an FA ratio greater than 1.10 remained viable. No region of interest with an rCBF ratio greater than 0.79 infarcted. CONCLUSIONS: Differences among mean ratios of three regions investigated were greatest for the rCBF ratio. The rCBF ratio may be the most useful parameter in differentiating viable tissue that is likely to infarct without intervention, from tissue that will survive despite hypoperfusion. ADC, DW intensity, FA, and rCBV ratios may provide adjunctive information. PMID- 12637295 TI - Cortical blood flow during cerebral vasospasm after aneurysmal subarachnoid hemorrhage: three-dimensional N-isopropyl-p-[(123)I]iodoamphetamine single photon emission CT findings. AB - BACKGROUND AND PURPOSE: The relationship between regional cerebral blood flow (rCBF) during cerebral vasospasm after subarachnoid hemorrhage (SAH) and angiographic vasospasm and the value of rCBF in predicting vasospasm and the prognosis are not fully delineated. Our aim was to investigate the changes in extent of vasospasm-induced decreased cortical rCBF on three-dimensional (3D) displays of single photon emission CT (SPECT) findings. The clinical usefulness of these assessments was analyzed. METHODS: In 58 cases of SAH, SPECT and digital subtraction angiography were performed on the same day, 5-9 days after SAH or within 24 hours after the onset of delayed ischemic neurologic deficit (DIND). Cerebral blood flow data were assessed by measuring the area of decreased cortical rCBF on 3D SPECT images. RESULTS: The area of decreased cortical rCBF on the 3D images was significantly increased in cases with DIND (P <.001), in cases with a large infarction due to vasospasm (P =.006), and in cases with a poor prognosis after vasospasm (P =.045). These increases were also related to the type of angiographic vasospasm; the greatest decrease in cortical rCBF occurred in the combined type (combination of the peripheral and proximal types) of vasospasm, followed by cases with the peripheral type, proximal type, and no angiographic vasospasm. In cases with DIND, patchy decreased cortical rCBF areas were seen before the onset of DIND. CONCLUSION: Combined-type vasospasm leads to reductions in CBF greater than those due to isolated peripheral or proximal vasospasm. Two-dimensional and mean-hemispheric CBF analyses are less sensitive for this change than is 3D SPECT. PMID- 12637296 TI - Subtraction helical CT angiography of intra- and extracranial vessels: technical considerations and preliminary experience. AB - BACKGROUND AND PURPOSE: Reported CT angiographic (CTA) subtraction methods are not simple, robust, or real time. We investigated a novel technique for semiautomated digital subtraction CTA of the intracranial and extracranial arteries. METHODS: Thirty patients underwent precontrast (low milliampere seconds) and postcontrast (pitch, 1.5; collimation, 1-2.5 mm) helical imaging with a vacuum-type head holder to facilitate image registration and minimize movement. A reconstructed three-dimensional model of the precontrast bone dataset was subtracted from the postcontrast dataset to produce subtracted maximum intensity-projection angiograms. Experienced (operator 1) and less-experienced (operator 2) staff performed the standard and subtraction reconstructions, and image generation time and quality (graded 1-5) were compared. A third operator blinded to the method assessed the hard-copy image quality. RESULTS: Image quality with subtraction postprocessing was significantly better with both operators (operator 1, mean improvement of 0.87 grade, median improvement of 1 grade, P <.001; operator 2, mean improvement of 0.63 grade, median improvement of 1 grade, P <.001). Hard-copy image quality was better with the subtraction method (operator 1, P >.001; operator 2, P <.001). Blood vessels at the base of the brain were better demonstrated on subtraction images in 13 of 14 examinations. For the less experienced operator, the reconstruction time was significantly less with the subtraction method than with the conventional method (mean, 7.5 vs 10.1 minutes; P =.001). CONCLUSION: When separation of the vasculature from bone is important and technically difficult, digital subtraction CTA offers a potential advantage. This semiautomated technique is fast and easy to learn, and variably experienced staff can use it. PMID- 12637297 TI - Assessment of the collateral function of the circle of Willis: three-dimensional time-of-flight MR angiography compared with transcranial color-coded duplex sonography. AB - BACKGROUND AND PURPOSE: Identification of the intracranial collaterals assists in identifying patients with severe occlusive disease of the internal carotid arteries who are at lower risk of transient ischemic attacks (TIAs) and stroke. We investigated the usefulness of MR angiography in identifying functional collaterals of the circle of Willis. METHODS: MR angiography of the circle of Willis was performed in 50 healthy volunteers. Visibility was used as the criterion to define the intracranial collaterals as being functional. Two observers independently assessed the MR angiograms. Results were compared with those of transcranial color duplex sonography (TCCD), and results of carotid compression tests were the standard of reference for the identification of functional intracranial collaterals. RESULTS: With MR angiograms, reviewer 1 achieved a sensitivity of 85%, a specificity of 81%, a positive predictive value of 95%, and a negative predictive value of 55%. Reviewer 2 achieved a sensitivity of 87%, a specificity of 67%, a positive predictive value of 92%, and a negative predictive value of 53%. Interobserver agreement on MR angiograms was moderate (kappa = 0.57, 95% confidence interval: 0.42, 0.72). CONCLUSION: Visible collaterals of the circle of Willis on MR angiograms are able to supply collateral flow in the presence of carotid artery obstruction. However, the low negative predictive value of MR angiography indicates that, if collaterals are not visible, supplementary TCCD investigation is required. PMID- 12637298 TI - Aneurysm clips: evaluation of magnetic field interactions and translational attraction by use of "long-bore" and "short-bore" 3.0-T MR imaging systems. AB - BACKGROUND AND PURPOSE: The use of 3.0-T MR systems is increasing worldwide. We evaluated magnetic field interactions and translational attraction for 32 aneurysm clips in association with exposure to "long-bore" and "short-bore" 3.0-T MR imaging systems. METHODS: Thirty-two different aneurysm clips were evaluated in this investigation. Each aneurysm clip was qualitatively evaluated for magnetic field interactions and quantitatively assessed for translational attraction by using the deflection angle test. The deflection angle tests were performed at the points of the highest spatial gradients for long-bore and short bore 3.0-T MR imaging systems. RESULTS: Seventeen of the 32 aneurysm clips showed positive magnetic field interactions. Deflection angles for the aneurysm clips were significantly (P <.001) higher on the short-bore (range, 0-18 degrees) compared with those recorded on the long-bore (range, 0-16 degrees) 3.0-T MR imaging system. Aneurysm clips made from commercially pure titanium and titanium alloy displayed no translational attraction (n = 15), whereas those made from stainless steel alloy, Phynox, and Elgiloy displayed positive deflection angles (n = 17). CONCLUSION: The 32 different aneurysm clips passed (angle <45 degrees) the deflection angle test by using the long- and short-bore 3.0-T MR imaging systems, indicating that they are safe for patients and other persons in these MR environments (ie, immediate area of MR imaging systems). However, only clips made from the titanium and titanium alloy are entirely safe for patients undergoing MR imaging procedures because of the total lack of magnetic field interactions. The remaining clips require characterization of magnetic field-induced torque. Because of possible differences in the points of the highest spatial gradients for different 3.0-T MR imaging systems, the results are specific to the imaging units and bore designs used in this investigation. PMID- 12637299 TI - Hyperacute subarachnoid hemorrhage on T2-weighted MR images. AB - We describe a case of hyperacute subarachnoid and intraventricular hemorrhage from a ruptured aneurysm, which occurred while the patient was undergoing MR imaging. Compared with CSF, the blood in the subarachnoid space had slightly lower signal intensity on T2*-weighted gradient-echo images and increased signal intensity on T2-weighted spin-echo images. This finding differs from the generally accepted MR appearance of intracranial hemorrhage and should be recognized to ensure proper patient care. PMID- 12637300 TI - Gabapentin therapy for amyotrophic lateral sclerosis: lack of improvement in neuronal integrity shown by MR spectroscopy. AB - BACKGROUND AND PURPOSE: Proton MR spectroscopy has revealed impaired neuronal integrity in the motor cortex of patients with amyotrophic lateral sclerosis (ALS). We hypothesized that the N-acetylaspartate (NAA)-creatine (Cr) ratios in the motor cortex and adjacent brain could reflect the therapeutic effectiveness of gabapentin (GBP) treatment in ALS. METHODS: Eight patients with ALS underwent MR spectroscopy before and 26.5 days +/- 8.8 after starting GBP. In 10 patients with ALS who were not treated with GBP, paired spectra were obtained 21.4 days +/ 7.2 apart. Fourteen healthy subjects underwent a single MR spectroscopic examination. The NAA/Cr ratio was measured in the precentral gyrus, the postcentral gyrus, the superior parietal lobule, the supplementary motor area, and the premotor cortex. RESULTS: The NAA/Cr ratio was decreased in the precentral and postcentral gyri of patients with ALS compared with healthy controls. In those with ALS, the change in the NAA/Cr ratio was not different between treated patients and untreated patients in any of the regions studied. CONCLUSION: No improvement in neuronal integrity was detected in motor and nonmotor cerebral regions after GBP treatment. This result agrees with that of prior investigations showing the equivocal clinical effectiveness of GBP for ALS and supports the validity of the NAA/Cr ratio as a surrogate of therapeutic effectiveness. PMID- 12637301 TI - Correlations of hippocampal atrophy and focal low-frequency magnetic activity in Alzheimer disease: volumetric MR imaging-magnetoencephalographic study. AB - BACKGROUND AND PURPOSE: Patients with Alzheimer disease (AD) have more low frequency activity on conventional EEG and increased focal magnetoencephalographic (MEG) dipole density (DD) in delta and theta bands. This activity concurs with atrophy and reduced metabolic and perfusion rates, particularly in temporoparietal structures. The relationship between functional and structural measures and their conjoined capability to improve the diagnosis of AD were assessed in this study. METHODS: Whole-head MEG recordings were obtained in 15 patients in whom the diagnosis of AD had been made and in 16 healthy control subjects during a resting condition. MR imaging volumetric data were also obtained; these included global cerebral, temporal lobe, and hippocampal volumes. RESULTS: DD in the delta and theta bands was enhanced in the AD group compared with the healthy control subjects. Slow-wave activity differed significantly between the groups in the temporoparietal regions of both hemispheres. Left hippocampal volume was correlated with left temporal and parietal delta DD and left temporal theta DD. A combination of left hippocampal volume and left temporal theta DD enabled correct classification in 87.1% of the patients with AD or control subjects. CONCLUSION: Results support the predominant role of temporoparietal hypofunction as defined by DD and hippocampal structural deficits shown on MR images in patients with AD. A multidisciplinary perspective of different techniques may improve our understanding of the disease and our diagnostic abilities. PMID- 12637302 TI - Lack of sex effect on brain activity during a visuomotor response task: functional MR imaging study. AB - BACKGROUND AND PURPOSE: As more individuals are enrolled in clinical functional MR imaging (fMRI) studies, an understanding of how sex may influence fMRI measured brain activation is critical. METHODS: We used fixed- and random-effects models to study the influence of sex on fMRI patterns of brain activation during a simple visuomotor reaction time task in the group of 26 age-matched men and women. We evaluated the right visual, left visual, left primary motor, left supplementary motor, and left anterior cingulate areas. RESULTS: Volumes of activations did not significantly differ between the groups in any defined regions. Analysis of variance failed to show any significant correlations between sex and volumes of brain activation in any location studied. Mean percentage signal-intensity changes for all locations were similar between men and women. A two-way t test of brain activation in men and women, performed as a part of random-effects modeling, showed no significant difference at any site. CONCLUSION: Our results suggest that sex seems to have little influence on fMRI brain activation when we compared performance on the simple reaction-time task. The need to control for sex effects is not critical in the analysis of this task with fMRI. PMID- 12637303 TI - Quantification of brain damage in cerebrotendinous xanthomatosis with magnetization transfer MR imaging. AB - BACKGROUND AND PURPOSE: Conventional MR imaging for quantification of brain damage in monitoring the evolution of cerebrotendinous xanthomatosis (CTX) has limitations. Magnetization transfer (MT) MR imaging is overcoming these limitations. Using MT MR imaging, we sought to quantify, in vivo, the extent of brain and cerebellar damage in patients with CTX, with the ulimate goal to investigate the magnitude of the correlation between MT MR imaging findings and clinical disability. METHODS: Conventional and MT MR images of the brain were obtained in nine patients with CTX and in 10 sex- and age-matched healthy volunteers. MT ratio histograms were derived of the whole brain, brain normal appearing white matter (NAWM), brain normal-appearing gray matter (NAGM), cerebellar NAWM, and cerebellar NAGM. Clinical disability was measured by using the Expanded Disability Status Scale (EDSS). RESULTS: Average MT ratio and peak heights of the whole brain, brain NAWM, and brain NAGM histograms in patients with CTX were significantly lower than the corresponding quantities in the control subjects. All cerebellar NAGM MT ratio histogram-derived metrics and average MT ratio of the cerebellar NAWM histogram in patients with CTX were also significantly lower than the corresponding quantities in the control subjects. Strong correlations were found between the EDSS score and a composite whole-brain MT ratio histogram score (r = 0.77, P <.01) and a composite brain white matter MT ratio histogram score (r = 0.71, P <.03). A strong correlation was also found between the cerebellar functional system score and a composite cerebellar NAWM score (r = 0.72, P <.02). CONCLUSION: The quantitative assessment of brain damage in patients with CTX with use of MT MR imaging can provide powerful measures of disease outcome. PMID- 12637304 TI - Early- and late-state subacute sclerosing panencephalitis: chemical shift imaging and single-voxel MR spectroscopy. AB - BACKGROUND AND PURPOSE: Subacute sclerosing panencephalitis (SSPE) is a rare, progressive, inflammatory neurodegenerative disease. Our aim was to determine the metabolic abnormalities of brain in early- and late-stage SSPE by using MR spectroscopy and to assess areas of involvement in the early stages when MR imaging findings were normal. METHODS: Children with stage II (n = 3) or III (n = 3) SSPE and 10 healthy, age-matched children underwent MR imaging, multivoxel MR spectroscopy, and short-echo single-voxel MR spectroscopy (SVS). Areas of involvement in the brain were determined with chemical shift imaging. For SVS, 2 x 2 x 2-cm voxels were placed in the frontal subcortical white matter (FSWM) and parieto-occipital white matter (POWM). N-acetylaspartate (NAA)/creatine (Cr), choline (Cho)/Cr, myo-inositol (Ins)/Cr, and NAA/Cho ratios were calculated. RESULTS: Comparisons of NAA/Cr, Cho/Cr, Ins/Cr and NAA/Cho ratios between patients and control subjects showed significant differences in FSWM and POWM (P <.01). In patients with SSPE, NAA/Cr ratios in POWM were significantly less than those in FSWM (P <.01). NAA/Cr ratios in patients with stage II SSPE and those in the control group were not significantly different; this may reflect the absence of neuronal loss. Decreased NAA/Cr, increased Cho/Cr and Ins/Cr ratios, and increased lactate and lipid peaks were found in patients with stage III SSPE. CONCLUSION: MR spectroscopy showed findings suggestive of inflammation in stage II and findings of demyelination, gliosis, cellular necrosis, and anaerobic metabolism in stage III. MR spectroscopy could be a promising technique for early diagnosis and treatment planning in cases of SSPE. PMID- 12637305 TI - MR imaging of autopsy-proved paraneoplastic limbic encephalitis in non-Hodgkin lymphoma. AB - We report the case of a 26-year-old man with precursor T-cell acute lymphoblastic leukemia who developed paraneoplastic limbic encephalitis that was diagnosed on the basis of MR imaging findings and was proved post mortem. In our MR imaging studies, fluid-attenuated inversion recovery images and diffusion-weighted echo planar images clearly depicted bilateral involvement of the medial temporal lobes and multifocal involvement of the brain, whereas T2-weighted turbo spin-echo images failed to show the changes. PMID- 12637306 TI - Black holes on MR images of the brain of patients with Bjork-Shiley heart valves: additional observation in three cases. AB - We report the cases of three patients with Bjork-Shiley convexo-concave heart valves with unusual black holes shown on cerebral MR images. For two patients, these findings were associated with fracture of the Bjork-Shiley convexo-concave heart valve, and for the third, with worn surfaces on the heart valve. Susceptibility changes shown by MR imaging suggest the possibility that these black holes may be associated with microscopic metallic particles, although, the exact cause of these black holes remains undetermined. PMID- 12637307 TI - Isolated intracranial Rosai-Dorfman disease (sinus histiocytosis with massive lymphadenopathy). AB - SUMMARY: Rosai-Dorfman disease (sinus histiocytosis with massive lymphadenopathy) rarely affects the intracranial region without involvement of other sites. We report the case of a 68-year-old woman with isolated Rosai-Dorfman disease of the frontal dura. She presented with a new onset seizure. Initial MR imaging showed subtle mild change in the left frontal region. During the ensuing 8 months, a dural mass made its symptomatic and definite MR imaging appearance in the same region. No extracranial lesion was present. PMID- 12637309 TI - Wegener granuloma in the fourth ventricle. AB - We report the case of a 57-year-old man with a known history of antineutrophil cytoplasmic antibody-positive Wegener granulomatosis with initial involvement of the upper and lower respiratory tract. MR imaging of the brain was performed because of new onset CNS symptoms (nausea, altered mental status). The images revealed a mass in the fourth ventricle. The mass diminished in size after systemic steroid treatment, as shown by follow-up MR imaging. The patient died in acute respiratory distress secondary to pneumonia. Histopathologic findings confirmed a Wegener granuloma of the fourth ventricle. PMID- 12637308 TI - CT and MR imaging of intracerebral amyloidoma: case report and review of the literature. AB - Intracerebral amyloidoma is the least common form of amyloid deposition in the brain. CT and MR imaging features in a case of pathologically proved cerebral amyloidoma are presented, and the available literature is reviewed. Typical imaging features of this entity include solitary or multiple supratentorial white matter masses that are hyperattenuated on nonenhanced CT. They have little or no mass effect on surrounding structures, extend medially up to the lateral ventricle wall, and have fine, irregular, enhancing margins. PMID- 12637310 TI - Guglielmi detachable coil treatment of ruptured saccular cerebral aneurysms: retrospective review of a 10-year single-center experience. AB - BACKGROUND AND PURPOSE: We retrospectively analyzed our results with Guglielmi detachable coils (GDCs) for the endovascular occlusion of acutely ruptured saccular cerebral aneurysms over 10 years. METHODS: Between 1991-2000, 83 patients (mean age, 56.1 years) with aneurysmal subarachnoid hemorrhage were treated with endovascular GDCs. Patients with aneurysms due to trauma or dissection and those with mycotic or fusiform aneurysms were excluded. Mean follow-up in survivors was 19.1 months, and the mean Hunt-Hess grade at admission was 2.2. Angiographic follow-up was performed in 93% of surviving patients (mean interval, 11.6 months). The basilar caput (34 patients) and anterior communicating artery complex (19 patients) were most commonly treated. RESULTS: Sixty-four patients (77%) had a Glasgow Outcome Scale score (GOS) of 4 or 5, nine (11%) had a score of 2 or 3, and 10 (12%) died. At follow-up, 24 patients (35%) had complete aneurysm occlusion, 18 (26%) had a dog-ear remnant, 24 (35%) had a residual neck, and two (3%) had residual aneurysm filling. No treated aneurysm rebled. Three patients required surgical repair after incomplete endovascular treatment. Two or more GDC occlusion procedures were required in 28 patients (34%). Major procedural complications occurred in two patients (2%), resulting in serious neurologic disability or death. CONCLUSION: Endovascular treatment of ruptured cerebral aneurysms with GDCs has low morbidity, and it facilitates good overall outcomes in patients after subarachnoid hemorrhage. The short-term effectiveness of GDC occlusion in preventing aneurysmal rebleeding was excellent. Durability of the treatment in preventing long-term rebleeding as compared with direct surgical clipping warrants further study. Advances in device technology and technique may improve future outcomes. PMID- 12637311 TI - North American clinical experience with the EKOS MicroLysUS infusion catheter for the treatment of embolic stroke. AB - BACKGROUND AND PURPOSE: We present early experience with the EKOS MicroLysUS infusion catheter for acute embolic stroke treatment in North America. This study was designed to demonstrate the safety of the device and to determine if sonography accelerates thrombolysis and improves clinical outcomes. METHODS: Fourteen patients aged 40-77 years with anterior- or posterior-circulation occlusion presented with cerebral ischemia 3-6 or 4-13 hours after symptom onset, respectively. Patients were treated with the catheter and simultaneous intraarterial thrombolysis. Procedural and clinical information, including time to lysis, degree of recanalization, National Institute of Health Stroke Scale (NIHSS) score, and modified Rankin Scale (mRS) score was recorded before treatment and afterward (immediately and at 24 hours, 1 week, 1 month, and 3 months). RESULTS: Ten patients presented with acute anterior-circulation emboli; four patients, with posterior-circulation emboli (NIHSS score, 9-23 [mean. 18.2] and 11-27 [mean, 18.75], respectively). Three deaths occurred at 24 hours: two from hemorrhage and one from cerebral swelling. Deaths also occurred at 1 week and 1 month after treatment. Thrombolysis in Myocardial Ischemia grade 2-3 flow was achieved in eight patients in the first hour. Average time to recanalization was 46 minutes. Mean NIHSS scores in eight of nine survivors at 90 days were 5 in the anterior-circulation group and 3 in the posterior-circulation group; mean mRS scores at 90 days were 2 and 3, respectively. No catheter-related adverse events occurred. CONCLUSION: Use of the EKOS MicroLysUS infusion catheter is feasible in the treatment of acute ischemic stroke. Further studies to evaluate its efficacy are warranted. PMID- 12637312 TI - Gas generation and clot formation during electrolytic detachment of Guglielmi detachable coils: in vitro observations and animal experiment. AB - BACKGROUND AND PURPOSE: Recent reports describe a high rate of thromboembolic events related to Guglielmi detachable coil (GDC) use in the treatment of cerebral aneurysms. The purpose of this study was to investigate electrolysis related changes of blood as a potential cause of thromboembolic complications associated with GDC use. METHODS: For in vitro observations, 15 GDCs (10 conventional coils and five insulated coils) were experimentally detached under microscopic observation. Three coils were detached in normal saline, five in human serum, and seven in heparinized human blood. For animal experiments, two coils were detached in two canine normal common carotid arteries with systemic heparinization. Immediately after detachment, the arteries were exposed, and clot formations were observed. RESULTS: Significant amounts of gas bubbles were observed in all in vitro observations; more were seen in conventional coils, which required longer detachment times, than in insulated coils. Gas generation started with the growth of tiny bubbles into larger ones. In insulated coils, gas was generated only at the detachment zone, and no difference between saline, serum, and blood environments was observed. During detachment within heparinized blood, clot formations of 2-3-mm diameter were observed at the detachment zones of insulated coils. Animal experiments showed clot formation at the detachment zone, and bubble entrapments around the clots were also found. CONCLUSION: The electrolytic detachment mechanism of the platinum coil can generate gas bubbles during the application of electric current. In association with electrothrombosis, this phenomenon may be a potential cause of thromboembolic complications during the treatment of cerebral aneurysms by use of GDCs. PMID- 12637313 TI - Extending polymerization time of polymethylmethacrylate cement in percutaneous vertebroplasty with ice bath cooling. AB - Currently available preparations of polymethylmethacrylate (PMMA) cement for percutaneous vertebroplasty have injectability times of 4-15 minutes. The potential for early polymerization requires procedures to be performed as fast as possible, sometimes with suboptimal results and waste of the cement. By cooling the PMMA mixture in an iced bath of sodium chloride solution, we can extend its injectability to over 2 hours and use one kit for the controlled treatment of multiple levels with successive needle placements. We have been using this technique since mid-1998, treating more than 600 vertebral bodies. PMID- 12637314 TI - Alterations in the intrinsic properties of the GPIbalpha-VWF tether bond define the kinetics of the platelet-type von Willebrand disease mutation, Gly233Val. AB - Platelet-type von Willebrand disease (PTVWD) is a bleeding disorder in which an increase of function mutation in glycoprotein Ibalpha (GPIbalpha), with respect to binding of von Willebrand factor (VWF), results in a loss of circulating high molecular weight VWF multimers together with a mild-moderate thrombocytopenia. To better ascertain the specific perturbations in adhesion associated with this disease state, we performed a detailed analysis of the kinetic and mechanical properties of tether bonds formed between PT-VWD platelets and the A1-domain of VWF. Results indicate that the GPIbalpha mutation, Gly233Val, promotes and stabilizes platelet adhesion to VWF at shear rates that do not support binding between the native receptor-ligand pair due to enhanced formation and increased longevity of the mutant tether bond (k0 off values for mutant versus native complex of 0.67 +/- 0.11 s-1 and 3.45 +/- 0.37 s-1, respectively). By contrast, the sensitivity of this interaction to an applied force, a measure of bond strength, was similar to the wild-type (WT) receptor. Although the observed alterations in the intrinsic properties of the GPIbalpha-VWF tether bond are comparable to those reported for the type 2B VWD, distinct molecular mechanisms may be responsible for these function-enhancing bleeding disorders, as interactions between the mutant receptor and mutant ligand resulted in a greater stability in platelet adhesion. We speculate that the enhanced cellular on-rate together with the prolongation in the lifetime of the mutant receptor-ligand bond contributes to platelet aggregation in circulating blood by permitting the formation of multiple GPIbalpha-VWF-A1 interactions. PMID- 12637315 TI - Expression of the candidate MCT-1 oncogene in B- and T-cell lymphoid malignancies. AB - Our laboratory has recently discovered a novel candidate oncogene, MCT-1, amplified in human T-cell lymphoma and mapped to chromosome Xq22-24. This region is amplified in a subset of primary B-cell non-Hodgkin lymphoma (NHL), suggesting that increased copy number of a gene(s) located in this region confers a growth advantage to some primary human lymphomas. We examined a diverse panel of lymphoid malignancies for the expression of MCT-1. We demonstrated that there are significantly increased levels of MCT-1 protein in a panel of T-cell lymphoid cell lines and in non-Hodgkin lymphoma cell lines. Furthermore, we identified a subset of primary diffuse large B-cell lymphomas that exhibited elevated levels of MCT-1 protein. Interestingly, all transformed follicular lymphomas in our study demonstrated elevated protein levels of MCT-1. There was no detectable MCT 1 protein in leukemic cells from patients with chronic lymphocytic leukemia or in any healthy lymphoid tissue examined. Lymphoid cell lines overexpressing MCT-1 exhibited increased growth rates and displayed increased protection against apoptosis induced by serum starvation when compared with matched controls. We found that MCT-1-overexpressing cells show constitutively higher levels of phosphorylated PKB/Akt protein, especially under serum starvation. Activation of survival pathways may be an additional function of the MCT-1 gene. Our data suggest that high levels of MCT-1 protein may be associated with a high-risk subset of lymphoid neoplasms and may further support the potential role of MCT-1 in promoting human lymphoid tumor development. PMID- 12637316 TI - IL-4 synergistically enhances both IL-2- and IL-12-induced IFN-gamma expression in murine NK cells. AB - Interleukin-4 (IL-4) is thought to influence T and natural killer (NK) cells by down-regulating T helper 1 (Th1)-type cytokines like interferon-gamma (IFN gamma). While investigating IL-4 regulation of IFN-gamma expression, we found that IL-4 synergized with IL-2 or IL-12 to enhance IFN-gamma production and mRNA expression in spleen-derived, IL-2-cultured NK cells, as well as negatively sorted fresh DX5+/CD3- NK cells albeit at lower levels. The positive effect of IL 4 on IL-2-induced IFN-gamma production was dependent upon signal transducer and activator of transcription 6 (Stat6) because this response was virtually abrogated in Stat6-/- mice. Notably, though, IL-12 plus IL-4 synergy on IFN-gamma expression was intact in Stat6-/- mice. In exploring possible molecular mechanisms to account for the synergistic effects of IL-4 on murine NK cells, we found that IL-2 plus IL-4 stimulation resulted in a modest increase in tyrosine phosphorylation of Stat5, while IL-12 plus IL-4 treatment resulted in a more substantial increase in tyrosine-phosphorylated Stat4. Finally, to identify regions of the IFN-gamma promoter that may be involved, NK cells from human IFN gamma promoter/luciferase transgenic mice were treated with cytokines. NK cells from proximal (-110 to +64) promoter region mice did not respond to cytokine stimulation; however, the intact -565 to +64 IFN-gamma promoter responded synergistically to IL-2 plus IL-4 and to IL-12 plus IL-4 in NK cells. These data demonstrate a role for IL-4 in enhancing IFN-gamma expression in murine NK cells that is partially dependent on Stat6 in IL-2 costimulation and completely independent of Stat6 in IL-12 costimulations. PMID- 12637317 TI - Result of high-dose imatinib mesylate in patients with Philadelphia chromosome positive chronic myeloid leukemia after failure of interferon-alpha. AB - Imatinib at 400 mg daily is effective in chronic-phase chronic myeloid leukemia (CML) after interferon failure, although only a few patients achieve a molecular remission. We investigated whether higher doses of imatinib may be more effective. Thirty-six patients with chronic-phase CML after failure on interferon alpha were treated with 400 mg imatinib twice daily. Median time from diagnosis was 25 months (range, 10-135 months); 4 patients (11%) had clonal evolution. All 11 patients with active disease achieved complete hematologic response. Excluding patients with fewer than 35% Ph-positive metaphases before the start of therapy, 19 (90%) of 21 evaluable patients achieved a major cytogenetic response. Of 27 evaluable patients, 24 (89%) achieved a complete cytogenetic response. Quantitative polymerase chain reaction was performed in bone marrow every 3 months. Of 32 evaluable patients, 18 (56%) showed BCR-ABL/ABL percentage ratios lower than 0.045%, including 13 (41%) with undetectable levels. With a median follow-up of 15 months, all patients were alive in chronic phase. Toxicities were similar to those reported with standard dose; 71% of patients continue to receive 600 mg or more of imatinib daily. In conclusion, high-dose imatinib induces complete cytogenetic responses in most patients with chronic-phase CML after interferon failure. This is accompanied by a high rate of molecular remission. PMID- 12637318 TI - Serine phosphorylation of STAT3 is essential for Mcl-1 expression and macrophage survival. AB - The Bcl-2 family member Mcl-1 is essential for macrophage survival. However, the mechanisms that contribute to the expression of Mcl-1 in these cells have not been fully characterized. The present study focused on the role of signal transducer and activator of transcription 3 (STAT3) in regulation of Mcl-1 in macrophages. Sodium salicylate (NaSal) treatment induced apoptotic cell death in primary human macrophages in a dose- and time-dependent fashion. Incubation with NaSal resulted in the loss of mitochondrial transmembrane potential, the release of cytochrome c and second mitochondria-derived activator of caspase/direct IAP binding protein with low pH of isoelectric point (pI) from the mitochondria, and the activation of caspases 9 and 3. Western blot analysis and reverse transcription-polymerase chain reaction demonstrated that NaSal down-regulated the expression of Mcl-1. Electrophoretic mobility shift assay and Western blot analysis for phosphorylated STAT3 demonstrated that STAT3 was constitutively activated in macrophages and that this STAT3 activation was suppressed by NaSal. The activation of STAT3 in macrophages was dependent on Ser727 phosphorylation, in the absence of detectable Tyr705 phosphorylation. Ectopic expression of STAT3 in murine RAW264.7 macrophages rescued the inhibition of Mcl-1 promoter-reporter gene activation and the cell death induced by NaSal treatment, while a dominant negative STAT3 resulted in cell death. To confirm its role in primary macrophages, STAT3 antisense (AS) oligodeoxynucleotides (ODNs) were employed. STAT3 AS, but not control, ODNs decreased STAT3 and Mcl-1 expression and resulted in macrophage apoptosis. These observations demonstrate that the STAT3-mediated expression of Mcl-1 is essential for the survival of primary human in vitro differentiated macrophages. PMID- 12637319 TI - Gene expression signatures in MLL-rearranged T-lineage and B-precursor acute leukemias: dominance of HOX dysregulation. AB - Rearrangements of the MLL locus, located on human chromosome 11q23, are frequent in both infant and therapy-related leukemias. Gene expression analysis of MLL rearranged B-precursor acute lymphoblastic leukemias (MLL B-ALLs) has identified these cases as a unique subtype of leukemia, characterized by the expression of genes associated with both lymphoid and myeloid hematopoietic lineages. Here we show that MLL fusions also generate a distinct genetic subtype of T-lineage ALL (MLL T-ALL), in which leukemic cells are characterized by an early arrest in thymocyte differentiation, with suggestive evidence of commitment to the gammadelta lineage. Interestingly, multiple genes linked to cell proliferation (eg, PCNA, MYC, CDK2, and POLA) were down-regulated in MLL-fusion samples, relative to those transformed by other T-ALL oncogenes (P <.000 001, Fisher exact test). Overall, MLL T-ALL cases consistently demonstrated increased levels of expression of a subset of major HOX genes--HOXA9, HOXA10, and HOXC6--and the MEIS1 HOX coregulator (P <.008, one-sided Wilcoxon test), a pattern of gene expression that was reiterated in MLL B-ALLs. However, expression of myeloid lineage genes, previously reported in MLL B-ALLs, was not identified in T-lineage cases with this abnormality, suggesting that myeloid gene dysregulation is dispensable in leukemic transformation mediated by MLL fusion proteins. Our findings implicate dysregulation of HOX gene family members as a dominant mechanism of leukemic transformation induced by chimeric MLL oncogenes. PMID- 12637320 TI - Membrane cholesterol regulates LFA-1 function and lipid raft heterogeneity. AB - Many surface receptors and signaling molecules are thought to associate with unique membrane microdomains termed lipid rafts. We examined the involvement of lipid rafts in the activation of leukocyte function-associated antigen-1 (LFA-1). Depletion or sequestration of cholesterol with methyl-beta-cyclodextrin (MCD) or filipin, respectively, strongly inhibited LFA-1-mediated adhesion of T-cell lines and primary T cells. This inhibition was reversed by cholesterol reconstitution. LFA-1 on T-cell lines was detected in cold Triton X-100-insoluble lipid rafts, which were disrupted by MCD or filipin treatment. However, no LFA-1 on primary T cells was detected in lipid rafts isolated by the same procedures, and these rafts were resistant to cholesterol depletion or sequestration. Association of LFA-1 with lipid rafts of primary T cells could be detected only when they were isolated with another nonionic detergent, Brij 35. Upon treatment with MCD, LFA-1 in Brij 35-insoluble lipid rafts partially shifted to nonraft fractions. T-cell lines were found to have a high level of cholesterol and a low level of ganglioside GM1, a common marker for lipid rafts, whereas primary T cells have a much lower level of cholesterol and a very high amount of GM1. Cross-linking of LFA-1 on primary T cells induced cocapping of cholesterol but not GM1. These results suggest that lipid rafts of T cells are heterogenous, and LFA-1 associates with a subset of lipid rafts containing a high level of cholesterol. This association seems to regulate LFA-1 functions, possibly by facilitating LFA 1 clustering. PMID- 12637321 TI - The proteasome inhibitor PS-341 sensitizes neoplastic cells to TRAIL-mediated apoptosis by reducing levels of c-FLIP. AB - Because of the pivotal role the proteasome plays in apoptosis, inhibitors of this enzyme, such as PS-341, provide a great opportunity for exploring synergy between proteasome inhibition and other apoptosis-inducing agents. Tumor necrosis factor related apoptosis-inducing ligand (TRAIL) can selectively induce apoptosis in tumor cells. In overnight assays, combinations of PS-341 and TRAIL were much more effective than either agent alone in promoting apoptosis of a murine myeloid leukemia, C1498, and a murine renal cancer, Renca. For C1498 cells, apoptosis sensitization by PS-341 affected neither the activity of nuclear factor kappaB (NF-kappaB) nor the levels of most antiapoptotic proteins. However, reductions in the antiapoptotic protein c-FLIP in response to PS-341 were observed in both C1498 and Renca cells. Treatment of normal bone marrow mixed with C1498 tumor cells for 18 hours with a combination of PS-341 and TRAIL resulted in a specific depletion of the tumor cells. Upon transfer to irradiated syngeneic recipient mice, mixtures treated with the PS-341 plus TRAIL combination resulted in enhanced long-term tumor-free survival of mice. These data therefore support the targeting of apoptotic pathways in tumor cells, using combinations of agents such as PS-341 and TRAIL that interact synergistically to preferentially promote tumor cell apoptosis. PMID- 12637322 TI - Human NK cell development in NOD/SCID mice receiving grafts of cord blood CD34+ cells. AB - Definition of the cytokine environment, which regulates the maturation of human natural killer (NK) cells, has been largely based on in vitro assays because of the lack of suitable animal models. Here we describe conditions leading to the development of human NK cells in NOD/SCID mice receiving grafts of hematopoietic CD34+ precursor cells from cord blood. After 1-week-long in vivo treatment with various combinations of interleukin (IL)-15, flt3 ligand, stem cell factor, IL-2, IL-12, and megakaryocyte growth and differentiation factor, CD56+CD3- cells were detected in bone marrow (BM), spleen, and peripheral blood (PB), comprising 5% to 15% of human CD45+ cells. Human NK cells of NOD/SCID mouse origin closely resembled NK cells from human PB with respect to phenotypic characteristics, interferon (IFN)-gamma production, and cytotoxicity against HLA class 1-deficient K562 targets in vitro and antitumor activity against K562 erythroleukemia in vivo. In the absence of growth factor treatment, CD56+ cells were present only at background levels, but CD34+CD7+ and CD34-CD7+ lymphoid precursors with NK cell differentiation potential were detected in BM and spleen of chimeric NOD/SCID mice for up to 5 months after transplantation. Our results demonstrate that limitations in human NK cell development in the murine microenvironment can be overcome by treatment with NK cell growth-promoting human cytokines, resulting in the maturation of IFN-gamma-producing cytotoxic NK cells. These studies establish conditions to explore human NK cell development and function in vivo in the NOD/SCID mouse model. PMID- 12637323 TI - ADAMTS13 activity in thrombotic thrombocytopenic purpura-hemolytic uremic syndrome: relation to presenting features and clinical outcomes in a prospective cohort of 142 patients. AB - Initial management of patients with thrombotic thrombocytopenic purpura-hemolytic uremic syndrome (TTP-HUS) is difficult because of lack of specific diagnostic criteria, high mortality without plasma exchange treatment, and risks of plasma exchange. Although severe ADAMTS13 (a disintegrin-like and metalloprotease with thrombospondin type 1 repeats) deficiency may be specific for TTP, the role of ADAMTS13 activity measurements for initial management decisions is unknown. ADAMTS13 was measured before beginning plasma exchange treatment in 142 (88%) of 161 consecutive patients with clinically diagnosed TTP-HUS with assignment to 1 of 4 categories: less than 5% (severe deficiency), 5% to 9%, 10% to 25%, and more than 25%. Eighteen (13%) of 142 patients had severe ADAMTS13 deficiency. Among 6 predefined clinical categories (stem cell transplantation, pregnant/postpartum, drug association, bloody diarrhea, additional/alternative disorder, idiopathic), severe deficiency occurred only among pregnant/postpartum (2 of 10) and idiopathic (16 of 48) patients. The presenting features and clinical outcomes of the 16 patients with idiopathic TTP-HUS who had severe ADAMTS13 deficiency were variable and not distinct from the 32 patients with idiopathic TTPHUS who did not have severe ADAMTS13 deficiency. Many patients in all ADAMTS13 activity categories apparently responded to plasma exchange treatment. Therefore, severe ADAMTS13 deficiency does not detect all patients who may be appropriately diagnosed with TTP-HUS and who may respond to plasma exchange treatment. PMID- 12637325 TI - Expression of hepcidin in hereditary hemochromatosis: evidence for a regulation in response to the serum transferrin saturation and to non-transferrin-bound iron. AB - Experimental data suggest the antimicrobial peptide hepcidin as a central regulator in iron homeostasis. In this study, we characterized the expression of human hepcidin in experimental and clinical iron overload conditions, including hereditary hemochromatosis. Using quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), we determined expression of hepcidin and the most relevant iron-related genes in liver biopsies from patients with hemochromatosis and iron-stain-negative control subjects. Regulation of hepcidin mRNA expression in response to transferrin-bound iron, non-transferrin-bound iron, and deferoxamine was analyzed in HepG2 cells. Hepcidin expression correlated significantly with serum ferritin levels in controls, whereas no significant up regulation was observed in patients with hemochromatosis despite iron-overload conditions and high serum ferritin levels. However, patients with hemochromatosis showed an inverse correlation between hepcidin transcript levels and the serum transferrin saturation. Moreover, we found a significant correlation between hepatic transcript levels of hepcidin and transferrin receptor-2 irrespective of the iron status. In vitro data indicated that hepcidin expression is down regulated in response to non-transferrin-bound iron. In conclusion, the presented data suggest a close relationship between the transferrin saturation and hepatic hepcidin expression in hereditary hemochromatosis. Although the causality is not yet clear, this interaction might result from a down-regulation of hepcidin expression in response to significant levels of non-transferrin-bound iron. PMID- 12637324 TI - GMCSF activates NF-kappaB via direct interaction of the GMCSF receptor with IkappaB kinase beta. AB - Granulocyte-macrophage colony-stimulating factor (GMCSF) has a central role in proliferation and differentiation of hematopoetic cells. Furthermore, it influences the proliferation and migration of endothelial cells. GMCSF elicits these functions by activating a receptor consisting of a ligand-specific alpha chain and a beta-chain, which is common for GMCSF, interleukin-3 (IL-3), and IL 5. It is known that various signaling molecules such as Janus kinase 2 or transcription factors of the signal transducer and activator of transcription (STAT) family bind to the common beta-chain and initiate signaling cascades. However, alpha-chain-specific signal transduction adapters have to be postulated given that IL-3, IL-5, and GMCSF induce partly distinct biologic responses. Using a yeast 2-hybrid system, we identified the alpha-chain of the GMCSF receptor (GMRalpha) as putative interaction partner of IkappaB kinase beta, one of the central signaling kinases activating the transcription factor nuclear factor kappaB (NF-kappaB). Using endogenous protein levels of endothelial cell extracts, we could verify the interaction by coimmunoprecipitation experiments. Fluorescence resonance energy transfer (FRET) microscopy confirmed the direct interaction of CFP-IKKbeta and YFPGMRalpha in living cells. Functional studies demonstrated GMCSF-dependent activation of IkappaB kinase activity in endothelial cells, degradation of IkappaB, and activation of NF-kappaB. Further biologic studies using GMCSF-dependent TF-1 cells indicated that GMCSF-triggered activation of NF-kappaB is important for cell survival and proliferation. PMID- 12637326 TI - Mutated VH genes and preferential VH3-21 use define new subsets of mantle cell lymphoma. AB - Mantle cell lymphoma (MCL) is believed to originate from a naive B cell. However, we recently demonstrated that a subset of MCL displayed mutated V(H) genes. We also reported restricted use of certain V(H) genes. To assess the prognostic impact of these new findings, we performed V(H) gene analysis of 110 patients, revealing that 18 (16%) patients had mutated and 92 (84%) patients had unmutated V(H) genes. Because the mutation rate was low in the mutated group (2.2%-6.7%), further investigation of the germline V(H) gene in T cells from 5 patients with mutated V(H) genes was carried out; results showed that the unrearranged V(H) gene was identical to the published sequence. These data confirm that the base pair substitutions within the rearranged V(H) genes represent hypermutations, and indicate germinal center exposure. However, V(H) gene mutation status did not correlate with prognosis because there was no difference in clinical outcome between the unmutated and mutated groups. The most frequently used V(H) genes were V(H)3-21 (21 patients) and V(H)4-34 (19 patients). A novel finding was that V(H)3-21(+) MCL almost exclusively expressed lambda light chains and displayed highly restricted use of the V(lambda)3-19 gene. V(H)3-21(+) patients had longer median survival than the remaining patients (53 vs 34 months; P =.03), but they tended to be younger at diagnosis. The combined use of V(H)3-21/V(lambda)3-19 suggests a possible role for antigen(s) in the pathogenesis of these tumors and indicates that V(H)3-21(+) patients constitute a new MCL entity. PMID- 12637327 TI - Ser727/Tyr701-phosphorylated Stat1 is required for the regulation of c-Myc, cyclins, and p27Kip1 associated with ATRA-induced G0/G1 arrest of U-937 cells. AB - All-trans retinoic acid (ATRA)-induced growth arrest of myeloid cells is associated with a sequential regulation of cyclins and cyclin-dependent kinase inhibitors (CKIs), which modulates the cell cycle machinery and inhibits the G1-S phase progression. ATRA treatment of myeloid cells induces up-regulation and tyrosine phosphorylation of Stat1, a member of the STAT (signal transducer and activator of transcription) transcription factor family that has been implicated in growth arrest in response to interferons. We have previously shown that ATRA induced cell cycle arrest is dependent on tyrosinephosphorylated Stat1. In this study, we show that there is a basal level of Stat1 Ser727 phosphorylation in U 937 cells, which is transiently increased in response to ATRA treatment. Using Stat1Ser727Ala-expressing sublines, we provide evidence that Ser727 phosphorylation of Stat1 is required for ATRA-induced growth arrest. To shed further light on the role of Stat1 in ATRA-induced cell cycle arrest, cyclin and CKI expression was analyzed during ATRA treatment in U-937 sublines expressing Stat1Ser727Ala and Stat1Tyr701Phe. Our results show that Ser727/Tyr701 phosphorylated Stat1 plays a key role as a prerequisite for the ATRA-induced down regulation of c-Myc; cyclins A, B, D2, D3, and E; and the simultaneous up regulation of p27Kip1, associated with arrest in the G0/G1 phase of the cell cycle. PMID- 12637328 TI - Sex significantly influences transduction of murine liver by recombinant adeno associated viral vectors through an androgen-dependent pathway. AB - A systematic evaluation of the influence of sex on transduction by recombinant adeno-associated viral vector (rAAV) indicated that transgene expression after liver-targeted delivery of vector particles was between 5- to 13-fold higher in male mice compared with female mice, irrespective of the proviral promoter or cDNA and mouse strain. Molecular analysis revealed that the rAAV genome was stably retained in male liver at levels that were 7-fold higher than those observed in females. Further, the sex difference in transduction was observed with AAV-2- and AAV-5-based vectors, which use distinct receptor complexes for infection. In concordance with the differences in AAV transduction, gel shift analysis with nuclear extracts derived from the liver of mice and humans revealed substantially higher binding of host nuclear protein to the rep-binding site (RBS) of AAV inverted terminal repeat (ITR) in males compared with females. Transduction efficiency and binding of nuclear protein to RBS was dramatically reduced in male mice by castration. In contrast, although oophorectomy did not significantly influence rAAV transduction, administration of 5alpha dihydrotestosterone, prior to gene transfer, increased stable hepatocyte gene transfer in females to levels observed in male mice, implying that androgens significantly influence hepatocyte gene transfer. Interestingly, sex did not have a significant effect on AAV gene transfer into nonhepatic tissue, indicating that there are distinct tissue- and sex-specific differences in the mechanisms responsible for efficient transduction with this vector. These results have significant implications for gene therapy of autosomal and acquired disorders affecting the liver. PMID- 12637329 TI - Multicenter phase 2 trial of thalidomide in relapsed/refractory multiple myeloma: adverse prognostic impact of advanced age. AB - Relapsed or refractory multiple myeloma has a poor outlook. Some patients respond to thalidomide; however, criteria for predicting response have not been conclusively identified. We initiated a prospective multicenter phase 2 trial in patients with relapsed/refractory myeloma using thalidomide up to the maximum dose, 800 mg/d. Interferon-alpha-2B (1.5-3.0 x 10(6) U, subcutaneously, 3 times per week) was added at week 12 if disease was responsive or stable. Patients intolerant of interferon continued thalidomide alone. Thalidomide with or without interferon was continued until disease progression. Objectives were to determine toxicity, response rate (RR), progression-free survival (PFS), and overall survival (OS) and to elucidate relevant prognostic factors. We enrolled 75 patients, with median age 64 years (range, 36-83 years). Median individual maximum-tolerated dose of thalidomide was 600 mg/d; 41% reached 800 mg/d. Overall RR was 28%, and 55% stable disease (SD). The only predictor for response was age 65 years or younger (38% versus 17%; P =.043). At 18 months median follow-up, the actuarial median PFS and OS were 5.5 and 14.6 months, respectively. Multivariate analysis for OS demonstrated age exceeding 65 years (median, 9.2 months versus longer than 26 months; P =.011), raised serum lactate dehydrogenase (P =.002), and raised serum creatinine (P =.007) predicted inferior outcomes. Nineteen patients received interferon. Ten discontinued owing to toxicity. Four of 12 patients who received interferon for longer than 4 weeks were converted from SD to partial response. Our findings confirm substantial activity of thalidomide in relapsed/refractory myeloma. Interferon may improve response in selected patients, but is often not tolerated. The inferior outcome demonstrated in those with the identified prognostic factors is important in planning management for such patients. PMID- 12637330 TI - Acquired FANCA dysfunction and cytogenetic instability in adult acute myelogenous leukemia. AB - Myelodysplastic and leukemic stem cell clones that evolve in children and adults with Fanconi anemia universally bear complex cytogenetic abnormalities. The abnormalities are generally recurring deletions or chromosomal loss and involve precisely the same chromosomes with the same frequency as has been described in marrow cells from patients with secondary acute leukemia induced by alkylating agents. Reasoning that acquired Fanconi anemia protein dysfunction might contribute to cytogenetic instability in secondary acute myelogenous leukemia (AML) cells, we analyzed leukemic cells bearing characteristic complex cytogenetic defects obtained from a 68-year-old man whose lymphoblasts showed no evidence of Fanconi anemia. Unlike the lymphoblasts, this myeloid leukemia cell line (UoC-M1) was hypersensitive to mitomycin-C (MMC) and diepoxybutane (DEB) and exhibited a marked decrease in nuclear FANCA, FANCG, and FANCD2-L. Retroviral transduction of FANCA significantly reduced MMC sensitivity but FANCF, FANCG, and FANCC did not. Overexpression of FANCA restored levels of both FANCA and FANCG, whereas overexpression of FANCG or FANCC did not restore FANCA levels. The molecular mass of cytoplasmic FANCA, FANCG, FANCC, and nuclear FANCD2 were normal. All exons of FANCA and FANCG were sequenced, and no mutations were found. We conclude that perturbations of as yet unidentified factors that govern the binding activity or intracellular localization of FANCA may promote cytogenetic instability and clonal progression in patients with AML who do not have Fanconi anemia. PMID- 12637331 TI - Reovirus oncolysis as a novel purging strategy for autologous stem cell transplantation. AB - Hematologic stem cell rescue after high-dose cytotoxic therapy is extensively used for the treatment of many hematopoietic and solid cancers. Gene marking studies suggest that occult tumor cells within the autograft may contribute to clinical relapse. To date purging of autografts contaminated with cancer cells has been unsuccessful. The selective oncolytic property of reovirus against myriad malignant histologies in in vitro, in vivo, and ex vivo systems has been previously demonstrated. In the present study we have shown that reovirus can successfully purge cancer cells within autografts. Human monocytic and myeloma cell lines as well as enriched ex vivo lymphoma, myeloma, and Waldenstrom macroglobulinemia patient tumor specimens were used in an experimental purging model. Viability of the cell lines or purified ex vivo tumor cells of diffuse large B-cell lymphoma, chronic lymphocytic leukemia, Waldenstrom macroglobulinemia, and small lymphocytic lymphoma was significantly reduced after reovirus treatment. Further, [35S]-methionine labeling and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of cellular proteins demonstrated reovirus protein synthesis and disruption of host cell protein synthesis as early as 24 hours. Admixtures of apheresis product with the abovementioned tumor cells and cell lines treated with reovirus showed complete purging of disease. In contrast, reovirus purging of enriched ex vivo multiple myeloma, Burkitt lymphoma, and follicular lymphoma was incomplete. The oncolytic action of reovirus did not affect CD34+ stem cells or their long-term colony-forming assays even after granulocyte colony-stimulating factor (G-CSF) stimulation. Our results indicate the ex vivo use of an unattenuated oncolytic virus as an attractive purging strategy for autologous stem cell transplantations. PMID- 12637333 TI - Human reconstituting hematopoietic stem cells up-regulate Fas expression upon active cell cycling but remain resistant to Fas-induced suppression. AB - The Fas receptor and its ligand have been implicated in mediating the bone marrow (BM) suppression observed in graft-versus-host disease and a number of other BM failure syndromes. However, previous studies have suggested that Fas is probably not expressed on human hematopoietic stem cells (HSCs), but up-regulated as a consequence of their commitment and differentiation, suggesting that progenitors or differentiated blood cells, rather than HSCs, are the targets of Fas-mediated suppression. The present studies confirm that candidate HSCs in human cord blood and BM lack constitutive expression of Fas, but demonstrate that Fas expression on CD34+ progenitor and stem cells is correlated to their cell cycle and activation status. With the use of recently developed in vitro conditions promoting HSC self-renewing divisions, Fas was up-regulated on virtually all HSCs capable of multilineage reconstituting nonobese diabetic/severe combined immunodeficiency (NOD-SCID) mice in vivo, as well as on long-term culture initiating cells (LTC-ICs). Similarly, in vivo cycling of NOD-SCID repopulating cells upon transplantation, resulted in up-regulation of Fas expression. However, repopulating HSCs expressing high levels of Fas remained highly resistant to Fas mediated suppression, and HSC function was compromised only upon coactivation with tumor necrosis factor. Thus, reconstituting human HSCs up-regulate Fas expression upon active cycling, demonstrating that HSCs could be targets for Fas mediated BM suppression. PMID- 12637332 TI - Stem cell factor protects erythroid precursor cells from chemotherapeutic agents via up-regulation of BCL-2 family proteins. AB - Suppression of red blood cell production is a common complication of chemotherapy, causing anemia in a significant number of cancer patients. We have evaluated the sensitivity of human hematopoietic progenitors and erythroid precursor cells to chemotherapeutic drugs and found that probasophilic erythroblasts represent the stage of erythroid differentiation more vulnerable to the cytotoxic effects of myelosuppressive agents. Stem cell factor (SCF) supports proliferation and survival of early hematopoietic cells by binding to the c-kit receptor. In unilineage erythropoietic culture of CD34+ progenitors, short-term pretreatment of immature erythroid precursors with SCF results in protection from apoptosis induced by chemotherapeutic agents and restores normal proliferation and differentiation after removal of the cytotoxic stimulus. The levels of drug induced caspase processing are significantly reduced in erythroblasts treated with SCF, indicating that activation of the c-kit receptor generates antiapoptotic signals acting before amplification of the caspase cascade. Accordingly, we found that SCF up-regulates Bcl-2 and Bcl-X L in erythroid precursors and that exogenous expression of these proteins protects erythroblasts from caspase activation and death induced by chemotherapeutic agents. These results suggest a possible mechanism for SCF-mediated protection of erythroid precursor cells from apoptosis and may contribute to devise new strategies for prevention and treatment of chemotherapy-induced anemia. PMID- 12637334 TI - Role of deferiprone in chelation therapy for transfusional iron overload. PMID- 12637335 TI - 2-methoxyestradiol alters cell motility, migration, and adhesion. AB - The effect of 2-methoxyestradiol, 2ME2, an endogenous metabolite of 17beta estradiol (E2), on cell growth and cytoskeletal functions in a BCR-ABL transformed cell line model was investigated. We determined the interaction of 2ME2 with STI571 (Gleevec, imatinib mesylate) in STI571 drug-sensitive and resistant cell lines. In cells expressing BCR-ABL, STI571 cooperated with 2ME2 in reducing cell growth, and STI571-resistant cells were sensitive to 2ME2 treatment. 2ME2 also inhibited growth of several cancer cell lines by a mechanism independent of BCR-ABL. BCR-ABL transformation leads to altered motility, increased adhesion, and spontaneous migration in different in vitro model systems. 2ME2 was found to specifically inhibit the spontaneous motility of BCRABL-transformed Ba/F3 cells and to change the morphology and volume of treated cells. Cells attached to fibronectin-coated surfaces showed a reduced number of filipodia and lamellipodia. In addition, 2ME2 significantly reduced BCRABL mediated adhesion to fibronectin. The spontaneous migration of BCR-ABL transformed cells through a transwell membrane also was found to be significantly decreased by 2ME2. Cytoskeletal changes were accompanied by alteration of tubulin formation, distinct from paclitaxel treatment. These results demonstrate that 2ME2 treatment of transformed cells strongly reduces cytoskeletal functions and may also be useful for the treatment of cancers with high metastatic potential. Combination of 2ME2 with other anticancer drugs may be beneficial to treatment of drug-resistant cancers. PMID- 12637336 TI - LDL containing apolipoprotein CIII is an independent risk factor for coronary events in diabetic patients. AB - OBJECTIVE: Triglyceride-rich lipoproteins that contain apolipoprotein CIII (apoCIII) are prominent in diabetic dyslipidemia. We hypothesized that these lipoproteins increase coronary disease risk in diabetic patients beyond that caused by standard lipid risk factors. METHODS AND RESULTS: Diabetic patients with previous myocardial infarction were followed for 5 years, and 121 who had a recurrent coronary event were matched to 121 who did not. VLDL and LDL that contained or did not contain apoCIII (CIII+ or CIII-) were prepared by immunoaffinity chromatography and ultracentrifugation. IDL was included in the LDL fraction. LDL CIII+, rich in cholesterol and triglyceride, was the strongest predictor of coronary events (relative risk [RR] 6.6, P<0.0001, for 4th versus 1st quartile). LDL CIII+ comprised 10% of total LDL. The main type of LDL, LDL CIII-, was less strongly predictive (RR 2.2, P=0.07). The increased risk associated with LDL CIII+ was unaffected by adjustment for plasma lipids, apoB, non-HDL cholesterol, or the other VLDL and LDL types. For VLDL CIII+, RR 0.5, P=0.07; for VLDL CIII-, RR 2.3, P=0.046. The presence of apolipoprotein E with CIII on VLDL and LDL did not affect risk. CONCLUSIONS: LDL with apoCIII strongly predicts coronary events in diabetic patients independently of other lipids and may be an atherogenic remnant of triglyceride-rich VLDL metabolism. PMID- 12637338 TI - Change in alpha1 HDL concentration predicts progression in coronary artery stenosis. AB - OBJECTIVE: We examined the effects of simvastatin-niacin and antioxidant vitamins on changes in high-density lipoprotein (HDL) subpopulations and alterations in coronary artery stenosis, as assessed by angiography. METHODS AND RESULTS: Lipids, lipoproteins, and HDL particles were measured on and off treatment in 123 subjects of the HDL-Atherosclerosis Treatment Study. Patients were assigned to 4 treatment groups, simvastatin-niacin, simvastatin-niacin-antioxidant vitamins, antioxidant vitamins, and placebo. Subjects were followed for 3 years on treatment and then for 2 months off treatment. Simvastatin-niacin significantly increased the 2 large apoA-I-containing HDL subpopulations, alpha1 and prealpha1, and significantly decreased the 2 smallest particles, prebeta1 and alpha3, compared with values obtained from the same patients off treatment. Adding antioxidant vitamins to the lipid-modifying agents blunted these effects (not significant). A significant negative correlation (r=-0.235; P<0.01) between the changes in alpha1 HDL particle concentration and coronary artery stenosis was noted. Subjects in the third tertile (157% increase in alpha1) had no progression of stenosis in the 3-year follow-up period, whereas subjects in the first tertile (15% decrease in alpha1) had an average of 2.1% increase in stenosis. CONCLUSIONS: Simvastatin-niacin therapy significantly increased the large apoA-I containing alpha1 HDL particles. This increase was significantly associated with less progression of coronary stenosis even after adjusting for traditional risk factors. PMID- 12637337 TI - Role of JNK, p38, and ERK in platelet-derived growth factor-induced vascular proliferation, migration, and gene expression. AB - OBJECTIVE: We investigated the comparative roles of mitogen-activated protein (MAP) kinases, including c-Jun NH2-terminal kinase (JNK), extracellular signal regulated kinase (ERK), and p38, in vascular smooth muscle cell (VSMC) proliferation, migration, and gene expression. METHODS AND RESULTS: VSMCs were infected with recombinant adenovirus containing dominant-negative mutants of ERK, p38, and JNK (Ad-DN-ERK, Ad-DN-p38, and Ad-DN-JNK, respectively) to specifically inhibit the respective MAP kinases and then stimulated with platelet-derived growth factor (PDGF)-BB. Ad-DN-ERK attenuated PDGF-BB-induced VSMC proliferation more potently than Ad-DN-p38 or Ad-DN-JNK, indicating the dominant role of ERK in VSMC proliferation. Ad-DN-ERK, Ad-DN-p38, and Ad-DN-JNK similarly inhibited PDGF induced VSMC migration. Ad-DN-ERK and Ad-DN-JNK suppressed PDGF-BB-induced downregulation of cyclin-dependent kinase inhibitor p27Kip1, whereas Ad-DN-p38 decreased PDGF-BB-induced upregulation of p21Cip1. Ad-DN-ERK inhibited PDGF-BB induced plasminogen activator inhibitor type-1 (PAI-1), monocyte chemoattractant protein-1, and transforming growth factor-beta1 expressions, Ad-DN-p38 blocked monocyte chemoattractant protein-1 and transforming growth factor-beta1 expression but not PAI-1, whereas Ad-DN-JNK suppressed only PAI-1 expression. Moreover, in vivo gene transfer of Ad-DN-p38 to rat carotid artery caused the inhibition of intimal hyperplasia by balloon injury, indicating the involvement of p38 in vascular remodeling in vivo. CONCLUSIONS: We propose that these 3 MAP kinases participate in vascular diseases via differential molecular mechanisms and are new therapeutic targets for treatment of vascular diseases. PMID- 12637339 TI - High density lipoprotein-induced angiogenesis requires the activation of Ras/MAP kinase in human coronary artery endothelial cells. AB - OBJECTIVE: Plasma high density lipoprotein (HDL) levels have been shown to be inversely correlated with coronary artery disease, but the mechanisms of the direct protective effect of HDL on endothelial cells (ECs) are not fully understood. In this study, we investigated the role of the HDL-mediated promotion of angiogenesis in human coronary artery ECs (HCECs). METHODS AND RESULTS: We developed an in vitro model of HCEC tube formation on a matrix gel. We optimized the maximum dose of HDL required to induce tube formation in initial experiments, in which the dose response showed that the maximum effective dose of HDL was 100 microg/mL. PD98059, an inhibitor of p42/44 mitogen-activated protein kinase (MAPK) activity, but not SB203580, an inhibitor of p38 MAPK activity, suppressed HDL-induced tube formation. Dominant-negative Ras N17 inhibited HDL-induced tube formation. HDL activated Ras according to a ras pull-down assay, and this effect was inhibited by pertussis toxin. Moreover, HDL activated phospho(p)-p42/44 MAPK, whereas Ras N17 blocked HDL-induced pp42/44 MAPK. CONCLUSIONS: These results indicate that HDL induced a potent signal through a Ras/MAPK pathway mediated by a pertussis toxin-sensitive G-protein coupled receptor to the angiogenic phenotype in HCECs. PMID- 12637340 TI - NAD(P)H oxidase mediates angiotensin II-induced vascular macrophage infiltration and medial hypertrophy. AB - OBJECTIVE: Our preliminary data suggested that angiotensin II (Ang II)-induced reactive oxygen species are involved in intercellular adhesion molecule-1 (ICAM 1) expression and leukocyte infiltration in the rat thoracic aorta. Other reports demonstrating reactive oxygen species-induced cell growth suggested a potential role of NAD(P)H oxidase in vascular hypertrophy. In the present study, we postulate that NAD(P)H oxidase is functionally involved in Ang II-induced ICAM-1 expression, macrophage infiltration, and vascular growth, and that oxidase inhibition attenuates these processes independently of a reduction in blood pressure. METHODS AND RESULTS: Rats were infused subcutaneously with vehicle or Ang II (750 microg/kg per day) for 1 week in the presence or absence of gp91 docking sequence (gp91ds)-tat peptide (1 mg/kg per day), a cell-permeant inhibitor of NAD(P)H oxidase. Immunohistochemical staining for ICAM-1 and ED1, a marker of monocytes and macrophages, showed that both were markedly increased with Ang II compared with vehicle and were reduced in rats receiving Ang II plus gp91ds-tat. This effect was accompanied by an Ang II-induced increase in medial hypertrophy that was attenuated by coinfusion of gp91ds-tat; however, gp91ds-tat had no effect on blood pressure. CONCLUSIONS: Ang II-enhanced NAD(P)H oxidase plays a role in the induction of ICAM-1 expression, leukocyte infiltration, and vascular hypertrophy, acting independently of changes in blood pressure. PMID- 12637341 TI - LOX-1, an oxidized LDL endothelial receptor, induces CD40/CD40L signaling in human coronary artery endothelial cells. AB - BACKGROUND: Despite increasing appreciation that atherogenesis involves participation of inflammatory cells, information on mediators of communication between different constituents of atherosclerotic plaque remain incomplete. We examined the role of LOX-1, a receptor for oxidized (ox) LDL, in the expression of CD40/CD40L in cultured human coronary artery endothelial cells (HCAECs). METHODS AND RESULTS: We observed that ox-LDL increased the expression of CD40 and CD40L in a concentration (10 to 80 microg/mL)- and time (1 to 24 hours)- dependent manner. These effects of ox-LDL were mediated by activation of LOX-1, because pretreatment of HCAECs with a blocking antibody to LOX-1 (JTX92) prevented the expression of CD40 and CD40L in response to ox-LDL (P<0.01). In parallel experiments, HCAECs were incubated with the protein kinase C (PKC) inhibitor bisindolylmaleimide I, and the cells were then exposed to ox-LDL. Both LOX-1 antibody and the PKC inhibitor inhibited PKC activation in response to ox LDL (P<0.01). The PKC inhibitor also blocked the effects of ox-LDL on the expression of CD40 and CD40L (P<0.01). In additional experiments, we found that it is the PKCalpha, but not PKCbeta and PKCgamma, isoform that mediated ox-LDL induced CD40 and CD40L upregulation. Further experiments showed that upregulation of CD40 mediated induction of proinflammatory genes, because CD40 antibody markedly reduced ox-LDL-induced TNF-alpha generation and P-selectin expression, whereas nonspecific mouse IgG had no effect. CONCLUSIONS: These findings indicate that ox-LDL through its receptor LOX-1 triggers the CD40/CD40L signaling pathway that activates the inflammatory reaction in HCAECs. These observations provide novel insight into ox-LDL-mediated inflammation in atherosclerosis. PMID- 12637342 TI - Integrin alphaIIbbeta3 and its antagonism. AB - AlphaIIbbeta3, the major membrane protein on the surface of platelets, is a member of the integrin family of heterodimeric adhesion receptors. The alphaIIb and beta3 subunits are each composed of a short cytoplasmic tail, a single transmembrane domain, and a large, extracellular region that consists of a series of linked domains. Recent structural analyses have provided insights into the organization of this and other integrins and how a signal is initiated at its cytoplasmic tail to transform the extracellular domain of alphaIIbbeta3 into a functional receptor for fibrinogen or von Willebrand factor to support platelet aggregation and thrombus formation. These functions of alphaIIbbeta3 have been targeted for antithrombotic therapy, and intravenous alphaIIbbeta3 antagonists have been remarkably effective in the setting of percutaneous coronary interventions, showing both short-term and long-term mortality benefits. However, the development of oral antagonists has been abandoned on the basis of excess of mortality in clinical trials, and the extension of therapy with existing alphaIIbbeta3 antagonists to broadly treat acute coronary syndromes has not fully met expectations. An in-depth understanding of how antagonists engage and influence the function of alphaIIbbeta3 and platelets in the context of the new structural insights may explain its salutary and potential deleterious effects. PMID- 12637343 TI - Human protease-activated receptor 1 expression in malignant epithelia: a role in invasiveness. AB - While protease-activated receptors (PARs) play a traditional role in vascular biology, they emerge with surprisingly new assignments in tumor biology. PAR1 expression correlates with the invasion properties of breast carcinoma, whereas human PAR1 antisense reduces their ability to migrate through Matrigel. Part of the molecular mechanism of PAR1 invasion involves the formation of focal contact complexes on PAR1 activation. PAR1 induces angiogenesis in animal models in vivo and exhibits an oncogenic phenotype of enhanced ductal complexity when overexpressed in mouse mammary glands. PMID- 12637344 TI - A genome-wide scan of pulmonary function measures in the National Heart, Lung, and Blood Institute Family Heart Study. AB - Spirometric measures of pulmonary function exhibited high heritability in the National Heart, Lung, and Blood Institute Family Heart Study. A genome scan of FEV1, FVC, and the ratio of FEV1/FVC was performed to identify chromosomal regions influencing these measures. The pulmonary traits were adjusted through multiple linear regression techniques for the effects of age, age2, body mass index, height, smoking status, and pack-years of smoking. The distribution of FEV1/FVC was transformed to account for nonnormality, and standardized residuals were used as the quantitative trait for variance component linkage analysis in GENEHUNTER (Whitehead Institute, Cambridge, MA). The genome scan identified regions on chromosomes 4 and 18 with logarithm of the odds favoring linkage (LOD) scores above 2.5, and these two chromosomes were further evaluated by incorporating additional marker genotyping. The FEV1/FVC ratio was linked to chromosome 4 around 28 centimorgans (cM; D4S1511) with a LOD score of 3.5, and the transformed ratio was linked to the same region with a LOD of 2.0. FEV1 and FVC were suggestively linked to regions on chromosome 18 with multipoint LOD scores of 2.4 for FEV1 and 1.5 for FVC at 31 cM (D18S843) and a LOD of 2.9 for FVC at 79 cM (D18S858). PMID- 12637345 TI - Norepinephrine release in brown adipose tissue remains robust in cold-exposed senescent Fischer 344 rats. AB - Near the end of life, old F344 rats undergo a transition, marked by spontaneous and rapidly declining function. Food intake and body weight decrease, and these rats, which we call senescent, develop severe hypothermia in the cold due in part to blunted brown fat [brown adipose tissue (BAT)] thermogenesis. We tested the hypothesis that this attenuation may involve diminished sympathetic signaling by measuring cold-induced BAT norepinephrine release in freely moving rats using linear microdialysis probes surgically implanted into interscapular BAT 24 and 48 h previously. In response to 2 h at 15 degrees C, senescent rats increased BAT norepinephrine release 6- to 10-fold but did not maintain homeothermy. This increase was comparable to that of old presenescent (weight stable) rats that did maintain homeothermy during even greater cold exposure (2 h at 15 degrees C followed by 1.5 h at 8 degrees C). Tail temperatures, an index of vasoconstrictor responsiveness to cold, exhibited similar cooling curves in presenescent and senescent rats. Thus cold-induced sympathetic signaling to BAT and tail vasoconstrictor responsiveness remain robust in senescent rats and cannot explain their cold-induced hypothermia. PMID- 12637346 TI - Effect of ANG II type I receptor antagonist and ACE inhibitor on vitamin D receptor-null mice. AB - We recently showed that vitamin D receptor (VDR) inactivation results in deregulated stimulation of the renin-angiotensin system (RAS). To address further the relation between RAS activation and the abnormalities in electrolyte and volume homeostasis, we studied the effect of the ANG II type I receptor antagonist losartan and the angiotensin-converting enzyme inhibitor captopril on VDR-null mice. Treatment with losartan or captopril normalized the water intake and urine excretion of VDR-null mice. However, the increase in salt excretion in VDR-null mice was not affected by either drug, suggesting that this abnormality is independent of the RAS. Northern blot and immunohistochemical analyses revealed that both drugs caused a drastic stimulation of renin expression in wild type and VDR-null mice, but renin expression remained much higher in the treated VDR-null mice than in the treated wild-type mice, suggesting that the ANG II feedback mechanism remains intact in the mutant mice. These data firmly established a causative relation between RAS overstimulation and the abnormal volume homeostasis in VDR-null mice and demonstrated that vitamin D repression of renin expression is independent of the ANG II feedback regulation in vivo. PMID- 12637347 TI - Ascorbate inhibits iNOS expression and preserves vasoconstrictor responsiveness in skeletal muscle of septic mice. AB - Inducible nitric oxide synthase (iNOS) expression in blood vessels contributes to the vascular hyporeactivity characteristic of sepsis. Our previous work demonstrated in vitro that ascorbate inhibits iNOS expression in lipopolysaccharide- and interferon-gamma-stimulated skeletal muscle endothelial cells (ECs) through an antioxidant mechanism. The present study evaluated in vivo the hypothesis that administration of ascorbate decreases oxidative stress, prevents endothelial iNOS expression, and improves vascular reactivity in septic skeletal muscle. Sepsis was induced in C57BL/6 mice by cecal ligation and puncture (CLP). Plasma nitrite and nitrate (NOx) levels were elevated by 6 h after CLP. Prior ascorbate bolus injection (200 mg/kg body wt iv) blocked the elevation of plasma NOx and abolished the expression of iNOS protein and activity in the septic skeletal muscle. We also demonstrated that iNOS mRNA determined by RT-PCR was induced in the microvascular ECs of the muscle at 3 h after CLP. This induction was attenuated by prior ascorbate administration. Ascorbate inhibition of iNOS expression was associated with decreased oxidant levels in the septic muscle. Moreover, ascorbate administration restored partially the baseline arterial pressure and preserved completely the microvascular constriction and arterial pressure responses to norepinephrine in CLP mice. These results suggest that early administration of ascorbate may be a valuable adjunct treatment of sepsis. PMID- 12637348 TI - Hypothermia preserves myocardial function and mitochondrial protein gene expression during hypoxia. AB - Hypothermia before and/or during no-flow ischemia promotes cardiac functional recovery and maintains mRNA expression for stress proteins and mitochondrial membrane proteins (MMP) during reperfusion. Adaptation and protection may occur through cold-induced change in anaerobic metabolism. Accordingly, the principal objective of this study was to test the hypothesis that hypothermia preserves myocardial function during hypoxia and reoxygenation. Hypoxic conditions in these experiments were created by reducing O2 concentration in perfusate, thereby maintaining or elevating coronary flow (CF). Isolated Langendorff-perfused rabbit hearts were subjected to perfusate (Po2 = 38 mmHg) with glucose (11.5 mM) and perfusion pressure (90 mmHg). The control (C) group was at 37 degrees C for 30 min before and 45 min during hypoxia, whereas the hypothermia (H) group was at 29.5 degrees C for 30 min before and 45 min during hypoxia. Reoxygenation occurred at 37 degrees C for 45 min for both groups. CF increased during hypoxia. The H group markedly improved functional recovery during reoxygenation, including left ventricular developed pressure (DP), the product of DP and heart rate, dP/dtmax, and O2 consumption (MVo2) (P < 0.05 vs. control). MVo2 decreased during hypothermia. Lactate and CO2 gradients across the coronary bed were the same in C and H groups during hypoxia, implying similar anaerobic metabolic rates. Hypothermia preserved MMP betaF1-ATPase mRNA levels but did not alter adenine nucleotide translocator-1 or heat shock protein-70 mRNA levels. In conclusion, hypothermia preserves cardiac function after hypoxia in the hypoxic high-CF model. Thus hypothermic protection does not occur exclusively through cold induced alterations in anaerobic metabolism. PMID- 12637349 TI - Heme oxygenase-mediated vasodilation involves vascular smooth muscle cell hyperpolarization. AB - Chronic hypoxia is associated with both blunted agonist-induced and myogenic vascular reactivity and is possibly due to an enhanced production of heme oxygenase (HO)-derived carbon monoxide (CO). However, the mechanism of endogenous CO-meditated vasodilation remains unclear. Isolated pressurized mesenteric arterioles from chronically hypoxic rats were administered the HO substrate heme l-lysinate (HLL) in the presence or absence of iberiotoxin, 1H [1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), ryanodine, or free radical spin traps (N-tert-butyl-alpha-phenylnitrone and 4,5-dihydroxy-1,3-benzenedisulfonic acid disodium salt). The effects of HLL administration on vascular smooth muscle (VSM) membrane potential were assessed in superior mesenteric artery strips in the presence and absence of zinc protoporphyrin IX or iberiotoxin. The vasodilatory responses to exogenous CO were assessed in the presence and absence of ODQ or iberiotoxin. HLL administration produced a dose-dependent vasodilatory response that was nearly eliminated in the presence of iberiotoxin. Neither ODQ, spin traps, nor ryanodine altered the vasodilatory response to HLL, although ODQ abolished the vasodilatory response to S-nitroso-N-acetyl-penicillamine. HLL administration produced a zinc protoporphyrin IX- and iberiotoxin-sensitive VSM cell hyperpolarization. Iberiotoxin and ODQ inhibited the vasodilatory response to exogenous CO. Thus the vasodilatory response to endogenous CO involves cGMP independent activation of VSM large-conductance Ca2+-activated K+ channels and does not likely involve the formation of Ca2+ sparks emanating from ryanodine sensitive stores. PMID- 12637350 TI - Determinants of left ventricular preload-adjusted maximal power. AB - Maximal left ventricular (LV) hydraulic power output (PWR(max)), corrected for preload as PWR(max)/(V(ed))(beta) (where V(ed) is the end-diastolic volume and beta is a constant coefficient), is an index of LV contractility. Whereas preload adjusted maximal power (PAMP) is usually calculated with beta = 2, there is uncertainty about the optimal value of beta (beta = 1 for the normal LV and 2 for the dilated LV). The aim of this work is to study the determining factors of beta. The data set consisted of 245 recordings (steady state and vena cava occlusion) in 10 animals in an ischemic heart pig model. The occlusion data yielded the slope (E(es); 2.01 +/- 0.77 mmHg/ml, range 0.71-4.16 mmHg/ml) and intercept (V(0); -11.9 +/- 22.6 ml; range -76 to 39 ml) of the end-systolic pressure-volume relation, and the optimal beta-factor (assessed by fitting an exponential curve through the V(ed)-PWR(max) relation) was 1.94 +/- 0.88 (range 0.29-4.73). The relation of beta with V(ed) was weak [beta = 0.60 + 0.02(V(ed)); r(2) = 0.20]. In contrast, we found an excellent exponential relation between V(0) and beta [beta = 2.16e(0.0189(V(0))), r(2) = 0.70]. PAMP, calculated from the steady-state data, was 0.64 +/- 0.40 mW/ml(2) (range 0.14-2.83 mW/ml(2)) with a poor correlation with E(es) (r = 0.30, P < 0.001). An alternative formulation of PAMP as PWR(max)/(V(ed) - V(0))(2), incorporating V(0), yielded 0.47 +/- 0.26 mW/ml(2) (range 0.09-1.42 mW/ml(2)) and was highly correlated with E(es) (r = 0.89, P < 0.001). In conclusion, correct preload adjustment of maximal LV power requires incorporation of V(0) and thus of data measured under altered loading conditions. PMID- 12637351 TI - Altered sinus nodal and atrioventricular nodal function in freely moving mice overexpressing the A1 adenosine receptor. AB - To investigate whether altered function of adenosine receptors could contribute to sinus node or atrioventricular (AV) nodal dysfunction in conscious mammals, we studied transgenic (TG) mice with cardiac-specific overexpression of the A1 adenosine receptor (A1AR). A Holter ECG was recorded in seven freely moving littermate pairs of mice during normal activity, exercise (5 min of swimming), and 1 h after exercise. TG mice had lower maximal heart rates (HR) than wild-type (WT) mice (normal activity: 437 +/- 18 vs. 522 +/- 24 beats/min, P < 0.05; exercise: 650 +/- 13 vs. 765 +/- 28 beats/min, P < 0.05; 1 h after exercise: 588 +/- 18 vs. 720 +/- 12 beats/min, P < 0.05; all values are means +/- SE). Mean HR was lower during exercise (589 +/- 16 vs. 698 +/- 34 beats/min, P < 0.05) and after exercise (495 +/- 16 vs. 592 +/- 27 beats/min, P < 0.05). Minimal HR was not different between genotypes. HR variability (SD of RR intervals) was reduced by 30% (P < 0.05) in TG compared with WT mice. Pertussis toxin (n = 4 pairs, 150 microg/kg ip) reversed bradycardia after 48 h. TG mice showed first-degree AV nodal block (PQ interval: 42 +/- 2 vs. 37 +/- 2 ms, P < 0.05), which was diminished but not abolished by pertussis toxin. Isolated Langendorff-perfused TG hearts developed spontaneous atrial arrhythmias (3 of 6 TG mice vs. 0 of 9 WT mice, P < 0.05). In conclusion, A1AR regulate sinus nodal and AV nodal function in the mammalian heart in vivo. Enhanced expression of A1AR causes sinus nodal and AV nodal dysfunction and supraventricular arrhythmias. PMID- 12637352 TI - Age-related functional effects linked to phosphatase activity in ventricular myocytes. AB - Conflicting reports exist regarding the influence of beta-adrenergic stimulation on the maximum velocity of shortening (Vmax) in ventricular myocytes. This may be due to an unrecognized effect of maturation. In the present study, the effects of beta-adrenergic receptor stimulation on myocytes from hearts of juvenile nonbred and young adult retired breeder female rats were compared. Ventricular myocytes from young adults had a beta-adrenergic-dependent increase in Vmax and Ca2+ dependent actomyosin ATPase that was not observed in myocytes from juveniles. Myocytes from young adults had both an increase in beta-myosin heavy chain (MHC) and higher basal serine/threonine phosphatase activity compared with juvenile rats. Additional studies established moderate increases in beta-MHC induced by hypothyroidism do not confer myocardial beta-adrenergic responsiveness, whereas inhibition of the higher phosphatase activity in myocytes from young adults blocks the age-dependent, beta-adrenergic-induced increase in cross-bridge cycling rates. We propose that the higher phosphatase activity of myocytes from young adults compared with juveniles allows for a greater functional response of the myocardium to beta-adrenergic stimulation. PMID- 12637353 TI - Adenosine and opioid receptor-mediated cardioprotection in the rat: evidence for cross-talk between receptors. AB - The relative roles of free-radical production, mitochondrial ATP-sensitive K+ (mitoKATP) channels and possible receptor cross-talk in both opioid and adenosine A1 receptor (A1AR) mediated protection were assessed in a rat model of myocardial infarction. Sprague-Dawley rats were subjected to 30 min of occlusion and 90 min of reperfusion. The untreated rats exhibited an infarct of 58.8 +/- 2.9% [infarct size (IS)/area at risk (AAR), %] at the end of reperfusion. Pretreatment with either the nonselective opioid receptor agonist morphine or the selective A1AR agonist 2-chloro-cyclopentyladenosine (CCPA) dramatically reduced IS/AAR to 41.1 +/- 2.2% and 37.9 +/- 5.5%, respectively (P < 0.05). Protection afforded by either morphine or CCPA was abolished by the reactive oxygen species scavenger N (2-mercaptopropionyl)glycine or the mitoKATP channel blocker 5-hydroxydecanoate. Both morphine- and CCPA-mediated protection were attenuated by the selective A1AR antagonist 1,3-dipropyl-8-cyclopentylxanthine and the selective delta1-opioid receptor (DOR) antagonist 7-benzylidenealtrexone. Simultaneous administration of morphine and CCPA failed to enhance the infarct-sparing effect of either agonist alone. These data suggest that both DOR and A1AR-mediated cardioprotection are mitoKATP and reactive oxygen species dependent. Furthermore, these data suggest that there are converging pathways and/or receptor cross-talk between A1AR- and DOR-mediated cardioprotection. PMID- 12637355 TI - Dobutamine responsiveness, PET mismatch, and lack of necrosis in low-flow ischemia: is this hibernation in the isolated rat heart? AB - The clinical hallmarks of hibernating myocardium include hypocontractility while retaining an inotropic reserve (using dobutamine echocardiography), having normal or increased [18F]fluoro-2-deoxyglucose-6-phosphate (18FDG6P) accumulation associated with decreased coronary flow [flow-metabolism mismatch by positron emission tomography (PET)], and recovering completely postrevascularization. In this study, we investigated an isolated rat heart model of hibernation using experimental equivalents of these clinical techniques. Rat hearts (n = 5 hearts/group) were perfused with Krebs-Henseleit buffer for 40 min at 100% flow and 3 h at 10% flow and reperfused at 100% flow for 30 min (paced at 300 beats/min throughout). Left ventricular developed pressure fell to 30 +/- 8% during 10% flow and recovered to 90 +/- 7% after reperfusion. In an additional group, this recovery of function was found to be preserved over 2 h of reperfusion. Electron microscopic examination of hearts fixed at the end of the hibernation period demonstrated a lack of ischemic injury and an accumulation of glycogen granules, a phenomenon observed clinically. In a further group, hearts were challenged with dobutamine during the low-flow period. Hearts demonstrated an inotropic reserve at the expense of increased lactate leakage, with no appreciable creatine kinase release. PET studies used the same basic protocol in both dual- and globally perfused hearts (with 250MBq 18FDG in Krebs buffer +/- 0.4 mmol/l oleate). PET data showed flow-metabolism "mismatch;" whether regional or global, 18FDG6P accumulation in ischemic tissue was the same as (glucose only) or significantly higher than (glucose + oleate) control tissue (0.023 +/- 0.002 vs. 0.011 +/- 0.002 normalized counts. s-1x g-1x min-1, P < 0.05) despite receiving 10% of the flow. This isolated rat heart model of acute hibernation exhibits many of the same characteristics demonstrated clinically in hibernating myocardium. PMID- 12637354 TI - Coronary blood flow regulation in exercising swine involves parallel rather than redundant vasodilator pathways. AB - In dogs, only combined blockade of vasodilator pathways [via adenosine receptors, nitric oxide synthase (NOS) and ATP-sensitive K+ (KATP) channels] results in impairment of metabolic vasodilation, which suggests a redundancy design of coronary flow regulation. Conversely, in swine and humans, blocking KATP channels, adenosine receptors, or NOS each impairs coronary blood flow (CBF) at rest and during exercise. Consequently, we hypothesized that these vasodilators act in parallel rather than in redundancy to regulate CBF in swine. Swine exercised on a treadmill (0-5 km/h), during control and after blockade of KATP channels (with glibenclamide), adenosine receptors [with 8-phenyltheophylline (8 PT)], and/or NOS [with Nomega-nitro-l-arginine (l-NNA)]. l-NNA, 8-PT, and glibenclamide each reduced myocardial O2 delivery and coronary venous O2 tension. These effects of l-NNA, 8-PT, and glibenclamide were not modified by simultaneous blockade of the other vasodilators. Combined blockade of KATP channels and adenosine receptors with or without NOS inhibition was associated with increased H+ production and impaired myocardial function. However, despite an increase in O2 extraction to >90% during administration of l-NNA + 8-PT + glibenclamide, vasodilator reserve could still be recruited during exercise. Thus in awake swine, loss of KATP channels, adenosine, or NO is not compensated for by increased participation of the other two vasodilator mechanisms. These findings suggest a parallel rather than a redundancy design of CBF regulation in the porcine circulation. PMID- 12637356 TI - Cardiac effects of burn injury complicated by aspiration pneumonia-induced sepsis. AB - Early fluid resuscitation, antimicrobials, early excision, and grafting have improved survival in the early postburn period; however, a significant incidence of pneumonia-related sepsis occurs after burn injury, often progressing to multiple organ failure. Recent studies have suggested that this initial injury (burn injury) primes the subject, producing an exaggerated response to a second insult, such as pneumonia-related sepsis. We developed an experimental animal model that included a third-degree burn over 40% of the total body surface area, followed by sepsis (intratracheal administration of Streptococcus pneumoniae, 4 x 106 colony-forming unit), which was produced either 48 or 72 h after burn injury in adult male rats. Hearts harvested after either burn alone, sepsis alone, or burn plus sepsis were used to assess either contractile function (Langendorff) or cardiomyocyte secretion of tumor necrosis factor-alpha, interleukin (IL)-1beta, IL-6, and IL-10 (ELISA). Experimental groups included the following: 1). sham (sham burn and no sepsis); 2). burn injury alone studied either 24, 48, or 72 h postburn; 3). pneumonia-related sepsis in the absence of burn injury; and 4). pneumonia-induced sepsis studied either 48 or 72 h after an initial burn injury. Burn injury alone (24 h) or sepsis alone produced myocardial contractile defects and increases in pro- and anti-inflammatory cytokine secretion by cardiomyocytes. Sepsis that occurred 48 h postburn exacerbated the cardiac contractile defects seen with either burn alone or sepsis alone. Sepsis that occurred 72 h postburn produced contractile defects resembling those seen in either burn alone or sepsis alone. In conclusion, our data suggest that burn injury primes the subject such that a second insult early in the postburn period produces significantly greater cardiac abnormalities than those seen with either burn alone or sepsis alone. PMID- 12637357 TI - Conducted dilations initiated by purines in arterioles are endothelium dependent and require endothelial Ca2+. AB - The signaling pathways underlying the regulation of vascular resistance by purines in intact microvessels and particularly in communication of remote vasomotor responses are unclear. One process by which remote regions of arterioles communicate is via transmission of signals axially along the vessel wall. In this study, we identified a pathway for local and conducted dilations initiated by purines. Adenosine (Ado) or ATP (bind P1 and P2 purinergic receptors, respectively) was micropipette applied to arterioles (maximum diameter approximately 40 microm) in the cheek pouch of anesthetized hamsters. Observations were made at the site of stimulation (local) or approximately 1200 microm upstream along the same vessel. P2 antagonists (pyridoxalphosphate-6 azophenyl-2',4'-disulfonic acid tetrasodium and suramin) inhibited local constriction to ATP, whereas local and upstream dilations were unaffected. In contrast, during inhibition of P1 receptors (with xanthine amine congener) the local constriction was unchanged, whereas both local and upstream dilations to ATP were inhibited. Hydrolysis of ATP to Ado is implicated in the dilator response as blocking 5'-ectonucleotidase (with alpha,beta-methyleneadenosine 5' diphosphate) attenuated ATP-induced dilations. After endothelium denudation, constriction to ATP was unchanged, but dilations to both ATP and Ado were inhibited, identifying endothelial cells (ECs) as the primary target for P1 mediated dilation. Purines increased EC Ca2+ locally and upstream. Chelation of EC Ca2+ (with BAPTA) abolished the local and upstream dilations to P1 receptor stimulation. Collectively, these data demonstrate that stimulation of P1 receptors on ECs produces a vasodilation that spreads to remote regions. There is an associated increase in EC Ca2+, which is a required signaling intermediate in the manifestation of both the local and axially communicated arteriolar dilations. PMID- 12637358 TI - Assessment of spatial inhomogeneities in intima media thickness along an arterial segment using its dynamic behavior. AB - To assess locally deviating structural and mechanical properties of arterial walls, the spatial variance in end-diastolic intima media thickness (IMT) and the change in IMT during the cardiac cycle (DeltaIMT) were determined along a short segment of the common carotid artery (15.86 mm), at 16 positions simultaneously. Intrasubject spatial inhomogeneities along the artery were revealed by a spatial variance significantly larger than the temporal variance over several beats. If differences between positions were confirmed, the extent of the inhomogeneity was obtained by comparison of IMT and DeltaIMT at each position with their spatial medians +/- the least-significant difference. Because no intersubject comparisons were necessary, a single session of several measurements was sufficient to assess inhomogeneities in the arterial wall properties of a subject, making the method independent of biological variability between subjects. The method was evaluated on 47 presumed healthy subjects (age range 21-75 yr). In 22 subjects, spatial inhomogeneities in DeltaIMT occurred (P < 0.05). In young subjects, DeltaIMT was locally decreased, i.e., in systole inhomogeneities were less compressed than their surrounding tissue. In older subjects, DeltaIMT was locally increased, i.e., the inhomogeneity was locally more compressed than its surrounding wall tissue. PMID- 12637359 TI - Cardiac kinin level in experimental diabetes mellitus: role of kininases. AB - Diabetes mellitus impairs the cardiac kallikrein-kinin system by reducing cardiac kallikrein (KLK) and kininogen levels, a mechanism that may contribute to the deleterious outcome of cardiac ischemia in this disease. We studied left ventricular (LV) function and bradykinin (BK) coronary outflow in buffer perfused, isolated working hearts (n = 7) of controls and streptozotocin (STZ) induced diabetic rats before and after global ischemia. With the use of selective kininase inhibitors, the activities of angiotensin I-converting enzyme, aminopeptidase P, and neutral endopeptidase were determined by analyzing the degradation kinetics of exogenously administered BK during sequential coronary passages. Basal LV function and coronary flow were impaired in STZ-induced diabetic rats. Neither basal nor postischemic coronary BK outflow differed between control and diabetic hearts. Reperfusion after 15 min of ischemia induced a peak in coronary BK outflow that was of the same extent and duration in both groups. In diabetic hearts, total cardiac kininase activity was reduced by 41.4% with an unchanged relative kininase contribution compared with controls. In conclusion, despite reduced cardiac KLK synthesis, STZ-induced diabetic hearts are able to maintain kinin liberation under basal and ischemic conditions because of a primary impairment or a secondary downregulation of kinin-degrading enzymes. PMID- 12637360 TI - Participation of intracellular Ca2+ stores in arteriolar conducted responses. AB - We examined the role played by intracellular Ca2+ stores in conducted vasomotor responses induced by phenylephrine (PE) in isolated hamster cremasteric arterioles. When applied briefly ( approximately 1 s) to isolated, cannulated arterioles by using pressure-pulse ejection from a micropipette, PE produced a strong local vasoconstriction and a very small biphasic conducted response (a small constriction followed by a dilation) that propagated several hundred micrometers along the vessel length. The conducted vasomotion was associated with a monophasic elevation of the endothelial cell intracellular Ca2+ concentration ([Ca2+]i) at the site of stimulation, as measured with the Ca2+ indicator fura 2. The Ca2+ pump inhibitor thapsigargin was used to limit filling of Ca2+ stores in smooth muscle and endothelial cells. Thapsigargin reduced baseline diameter and elicited a strong dilator component at the local site while enhancing both the constrictor and dilator components of the PE-induced conducted response. The enhanced conducted constrictor component induced by thapsigargin was mimicked by extraluminal application of tetraethylammonium or charybdotoxin but not by iberiotoxin, apamin, glibenclamide, barium, or 4-aminopirydine. Thapsigargin increased the estimated basal endothelial cell [Ca2+]i by approximately 60 nM and converted the PE-induced change in [Ca2+]i from monotonic to biphasic with a late elevation of [Ca2+]i above baseline that coincided with the increased dilatory component of the conducted response. Luminal application of charybdotoxin plus apamin significantly reduced the dilatory component of the conducted response. These results indicate that intracellular Ca2+ stores play a dynamic role in regulating conducted vasomotor responses apparently through modulation of KCa channels in both cell types. PMID- 12637361 TI - Exercise restores beta-adrenergic vasorelaxation in aged rat carotid arteries. AB - Aging is associated with alterations in beta-adrenergic receptor (beta-AR) signaling and reduction in cardiovascular responses to beta-AR stimulation. Because exercise can attenuate age-related impairment in myocardial beta-AR signaling and function, we tested whether training could also exert favorable effects on vascular beta-AR responses. We evaluated common carotid artery responsiveness in isolated vessel ring preparations from 8 aged male Wistar-Kyoto (WKY) rats trained for 6 wk in a 5 days/wk swimming protocol, 10 untrained age matched rats, and 10 young WKY rats. Vessels were preconstricted with phenylephrine (10-6 M), and vasodilation was assessed in response to the beta-AR agonist isoproterenol (10-10-3 x 10-8 M), the alpha2-AR agonist UK-14304 (10-9-10 6 M), the muscarinic receptor agonist ACh (10-9-10-6 M), and nitroprusside (10-8 10-5 M). beta-AR density and cytoplasmic beta-AR kinase (beta-ARK) activity were tested on pooled carotid arteries. beta-ARK expression was assessed in two endothelial cell lines from bovine aorta and aorta isolated from a 12-wk WKY rat. beta-AR, alpha2-AR, and muscarinic responses, but not that to nitroprusside, were depressed in untrained aged vs. young animals. Exercise training restored beta-AR and muscarinic responses but did not affect vasodilation induced by UK-14304 and nitroprusside. Aged carotid arteries showed reduced beta-AR number and increased beta-ARK activity. Training counterbalanced these phenomena and restored beta-AR density and beta-ARK activity to levels observed in young rat carotids. Our data indicate that age impairs beta-AR vasorelaxation in rat carotid arteries through beta-AR downregulation and desensitization. Exercise restores this response and reverts age-related modification in beta-ARs and beta-ARK. Our data support an important role for beta-ARK in vascular beta-AR vasorelaxation. PMID- 12637362 TI - 17beta-estradiol decreases vascular tone in cerebral arteries by shifting COX dependent vasoconstriction to vasodilation. AB - We have previously shown that estrogen treatment increases cerebrovascular cyclooxygenase-1, prostacyclin synthase, and production of prostacyclin. Therefore, vascular tone and prostanoid production were measured to investigate functional consequences of estrogen exposure. Middle cerebral arteries were isolated from ovariectomized female Fischer-344 rats with or without chronic in vivo 17beta-estradiol treatment. In vivo 17beta-estradiol treatment increased cerebral artery diameter; functional endothelium was required for expression of these differences. The nonspecific cyclooxygenase inhibitor indomethacin constricted, whereas arachidonic acid dilated, cerebral arteries from estrogen treated animals. Estrogen exposure increased production of prostacyclin by cerebral arteries. Conversely, in estrogen-deficient animals, indomethacin dilated and arachidonic acid constricted cerebral blood vessels. This correlated with vasorelaxation following inhibition of the thromboxane-endoperoxide receptor with SQ-29548 but not after selective blockade of thromboxane synthase with furegrelate, suggesting prostaglandin endoperoxide (i.e., PGH2) activity. Removal of the endothelium or selective blockade of cyclooxygenase-1 with SC-560 abolished estrogen-mediated differences in the effects of arachidonate on vessel diameter and on prostacyclin production by cerebral arteries. These data suggest 17beta-estradiol decreases cerebrovascular tone by shifting the primary end product of the endothelial cyclooxygenase-1 pathway from the constrictor prostaglandin PGH2 to the vasodilator prostacyclin. These effects of estrogen may contribute to the heightened thromboresistance and enhanced cerebral blood flow documented in pre-versus postmenopausal women. PMID- 12637363 TI - Role of bradykinin in angiotensin-converting enzyme knockout mice. AB - Angiotensin-converting enzyme (ACE) plays a central role in the renin-angiotensin system. Whereas ACE is responsible for the production of angiotensin II, it is also important in the elimination of bradykinin. Constitutively, the biological function of bradykinin is mediated through the bradykinin B(2) receptor. ACE knockout mice have a complicated phenotype including very low blood pressure. To investigate the role of bradykinin in the expression of the ACE knockout phenotype, we bred B(2) receptor knockout mice with ACE knockout mice, thus generating a line of mice deficient in both the B(2) receptor and ACE. Surprisingly, these mice did not differ from ACE knockout mice in blood pressure, urine concentrating ability, renal pathology, and hematocrit. Thus abnormalities of bradykinin accumulation do not play an important role in the ACE knockout phenotype. Rather, this phenotype appears due to the defective production of angiotensin II. PMID- 12637364 TI - Central role of connexin40 in the propagation of electrically activated vasodilation in mouse cremasteric arterioles in vivo. AB - When a short segment of arteriole is stimulated, vasomotor responses spread bidirectionally along the vessel axis purportedly via gap junctions. We used connexin40 knockout (Cx40-/-) mice to study vasomotor responses induced by 10 second trains of electrical stimulation (30 Hz, 1 ms, 30 to 50 V) in 2nd or 3rd order arterioles of the cremaster muscle. Measurements were made at the stimulation site (local) and at conducted sites (500, 1000, and 2000 microm upstream). In wild-type (Cx40+/+) animals, electrical stimulation evoked a local vasoconstriction and a conducted vasodilation that spread very rapidly along the vessel length without detectable decay. In Cx40-/- mice, the conducted dilation was converted into either vasoconstriction or a slowly developing vasodilation that decayed along the vessel length. Tetrodotoxin (TTX, 1 micromol/L) had no effect on the local vasoconstriction in either Cx40+/+ or Cx40-/- mice, but enhanced the conducted vasodilation in Cx40+/+ animals. In Cx40-/- mice, TTX abolished the conducted vasoconstriction when present and revealed a small vasodilation that decayed with distance. In the group of Cx40-/- mice in which electrical stimulation elicited a conducted vasodilation, TTX had no effect. Immunocytochemistry revealed Cx40 only in the endothelial layer of arterioles from Cx40+/+ mice and complete elimination of this connexin in the Cx40-/- animals. These results indicate that focal current stimulation causes vasoconstriction by a combination of perivascular nerve stimulation and smooth muscle activation. Moreover, electrical stimulation activates a nonneuronal, Cx40 dependent vasodilator response that spreads along the vessel length without decay. PMID- 12637366 TI - The endothelial glycocalyx protects against myocardial edema. AB - Myocardial tissue edema attributable to increased microvascular fluid loss contributes to cardiac dysfunction after myocardial ischemia, cardiopulmonary bypass, hypertension, and sepsis. Recent studies suggest that carbohydrate structures on the luminal surface of microvascular endothelium are essential to prevent tissue edema. We carefully preserved these structures for visualization with electron microscopy, revealing that the rat myocardial capillary endothelial surface is coated with a 0.2- to 0.5-microm-thick carbohydrate layer and that its degradation instantly results in notable myocardial tissue edema. PMID- 12637365 TI - A novel, voltage-dependent nonselective cation current activated by insulin in guinea pig isolated ventricular myocytes. AB - Insulin regulates cardiac metabolism and function by targeting metabolic proteins or voltage-gated ion channels. This study provides evidence for a novel, voltage dependent, nonselective cation channel (NSCC) in the heart. Under voltage clamp at 37 degrees C and with major known conductances blocked, insulin (1 nmol/L to 1 micromol/L) activated an outwardly rectifying current (Iinsulin) in guinea pig ventricular myocytes. Iinsulin could be carried by Cs+, K+, Li+, and Na+ ions but not by NMDG+. It was inhibited by the NSCC blockers gadolinium and SKF96365 but not flufenamic acid. Iinsulin was largely blocked by the insulin receptor tyrosine kinase inhibitor HNMPA-(AM)3 and by the phospholipase C inhibitor U73122 but not by its inactive analogue U73433. Staurosporine, a potent blocker of protein kinase C, did not prevent the activation of Iinsulin. Application of an analogue of diacylglycerol, 1-oleoyl-2-acetyl-sn-glycerol, mimicked the effect of insulin. This activated an outwardly rectifying NSCC that could be carried by Cs+, K+, Li+, or Na+ and that was blocked by gadolinium but not by flufenamic acid or staurosporine. We conclude that the intracellular pathway leading to activation of this novel cardiac NSCC involves phospholipase C, is protein kinase C-independent, and may depend on direct channel activation by diacylglycerol. PMID- 12637367 TI - Chronic SR Ca2+-ATPase inhibition causes adaptive changes in cellular Ca2+ transport. AB - Phospholamban, the critical regulator of the cardiac SERCA2a Ca2+ affinity, is phosphorylated at Ser16 and Thr17 during beta-adrenergic stimulation (eg, isoproterenol). To assess the impact of nonphosphorylatable phospholamban, a S16A, T17A double-mutant (DM) was introduced into phospholamban knockout mouse hearts. Transgenic lines expressing DM phospholamban at levels similar to wild types (WT) were identified. In vitro phosphorylation confirmed that DM phospholamban could not be phosphorylated, but produced the same shift in EC50 of SERCA2a for Ca2+ as unphosphorylated WT phospholamban. Rates of basal twitch [Ca2+]i decline were not different in DM versus WT cardiomyocytes. Isoproterenol increased the rates of twitch [Ca2+]i decline in WT, but not DM myocytes, confirming the prominent role of phospholamban phosphorylation in this response. Increased L-type Ca2+ current (ICa) density, with unaltered characteristics, was the major compensation in DM myocytes. Consequently, the normal beta-adrenergic induced increase in ICa caused larger dynamic changes in absolute ICa density. Isoproterenol increased Ca2+ transients to a comparable amplitude in DM and WT. There were no changes in myofilament Ca2+ sensitivity, or the expression levels and Ca2+ removal activities of other Ca2+-handling proteins. Nor was there evidence of cardiac remodeling up to 10 months of age. Thus, chronic inhibition of SERCA2a by ablation of phospholamban phosphorylation (abolishing its adrenergic regulation) results in a unique cellular adaptation involving greater dynamic ICa modulation. This ICa modulation may partly compensate for the loss in SERCA2a responsiveness and thereby partially normalize beta-adrenergic inotropy in DM phospholamban mice. PMID- 12637368 TI - Ets-1 and Ets-2 transcription factors are essential for normal coronary and myocardial development in chicken embryos. AB - In the development of a functional myocardium and formation of the coronary vasculature, epicardium-derived cells play an essential role. The proepicardial organ contributes to the developing coronary system by delivering mural cells to the endothelium-lined vessels. In search of genes that regulate the behavior of (pro)epicardial cells, the Ets-1 and Ets-2 transcription factors stand out as strong candidates. In the present study, the hypothesis that Ets transcription factors have a role in proper coronary and myocardial development was tested via antisense technology, by targeting Ets-1 and Ets-2 mRNAs to downregulate protein expression in chicken embryos. The results suggest that hereby the development of the coronary system is hampered, primarily by defects in the process of epithelial-mesenchymal transformation of the mesothelia of the primary and secondary heart fields. This was indicated by a lack of periarterial and epicardial mesenchyme, of peripheral coronary smooth muscle cells, and changes in myocardial morphology. A defect in myocardial perfusion caused by the absence of one or both coronary ostia seems to be "solved" by the development of numerous small fistulae connecting the ventricular lumen with the subepicardially located coronary vessels. The presence of coronary vascular aberrations in the antisense Ets phenotype enabled us for the first time to study abnormal coronary development in a model that is not lethal to the embryo. PMID- 12637369 TI - Functional roles of the rho/rho kinase pathway and protein kinase C in the regulation of cerebrovascular constriction mediated by hemoglobin: relevance to subarachnoid hemorrhage and vasospasm. AB - Although there is evidence that the Rho/Rho kinase pathway and protein kinase C (PKC) are involved in the development of cerebral vasospasm, the mechanism by which subarachnoid hemorrhage (SAH) activates these pathways is unclear. A large body of evidence points to oxyhemoglobin (OxyHb) as a major causative component of blood clot responsible for vasospasm. Therefore, the present studies were conducted to explore whether the Rho/Rho kinase and PKC may be involved in a sustained vasoconstriction induced by OxyHb in cerebral arteries. OxyHb evoked sustained vasoconstriction in the endothelium-denuded rabbit basilar arteries, which was reversed by the selective inhibitors of Rho kinase, Y-27632, and HA 1077, with the IC50 values of 0.26+/-0.02 and 0.74+/-0.1 micromol/L, respectively. In quiescent cerebrovascular smooth muscle (CVSM) cells, OxyHb induced Rho translocation, as assessed by immunoblotting, with a time course, which paralleled the contractile action of OxyHb. Rho translocation was also observed in intact arteries stimulated with OxyHb for 24 hours (219%) and 48 hours (160%). The increase in Rho translocation was fully inhibited by GGTI-297, an inhibitor of Rho prenylation. OxyHb also caused significant translocation of both PKCalpha and PKCepsilon (P<0.01), which was maximal at the time corresponding to maximal tension developed in response to OxyHb. Ro-32-0432, an inhibitor of PKC, attenuated vasoconstriction mediated by OxyHb in basilar artery. These results show, for the first time, that OxyHb-mediated signaling in CVSM utilizes the Rho/Rho kinase and PKC-based mechanisms. PMID- 12637370 TI - Factor associated with neutral sphingomyelinase activation and its role in cardiac cell death. AB - Generation of proapoptotic sphingolipids by neutral sphingomyelinase activation is an early response to hypoxia/reoxygenation (HR) in cardiomyocytes. Factor associated with neutral sphingomyelinase activation (FAN) mediates activation of sphingomyelinase and subsequent apoptosis. However, the participation of FAN in HR-induced cardiomyocyte cell death has not been elucidated. We therefore investigated the expression and role of FAN in rat cardiomyocytes. A cDNA was isolated from rat heart encoding putative rat FAN. Reverse transcriptase polymerase chain reaction, immunoelectron microscopy, and immunofluorescence demonstrated for the first time the expression of FAN specifically in rat cardiomyocytes. FAN expression was confirmed by the finding that expression of a dominant-negative FAN almost completely abrogated HR-induced cell death, whereas overexpression of wild-type FAN led to an increase. Treatment of FAN and dominant negative FAN--expressing cells with C2-ceramide produced substantial cell death, indicating dominant-negative FAN exerts its protective action by interfering with the activation of the sphingolipid cascade. Taking these results together, we conclude that FAN is a previously undescribed and important HR signaling component in the heart and that inhibition of FAN may provide a novel intervention point for reducing ischemia/reperfusion injury. PMID- 12637371 TI - Sildenafil induces delayed preconditioning through inducible nitric oxide synthase-dependent pathway in mouse heart. AB - Sildenafil citrate (Viagra) is the most widely used drug for treating erectile dysfunction in men. We recently demonstrated that it induces potent protective effects against ischemia-reperfusion (I-R) injury in rabbit hearts through the opening of mitochondrial ATP-dependent K+ channels. In the present study, we investigated the role of the NO-dependent signaling pathway in delayed cardioprotection by sildenafil. Adult male ICR mice were treated with saline or sildenafil (0.7 mg/kg IP) 24 hours before global I-R in the Langendorff mode. Infarct size was reduced from 27.6+/-3.3% in saline-treated control mice to 6.9+/ 1.2% in sildenafil-treated mice (mean+/-SEM, P<0.05) without compromising cardiac function. Reverse transcription-polymerase chain reaction revealed a transient increase in endothelial and inducible NO synthase (eNOS and iNOS, respectively) mRNA in sildenafil-treated mice, peaking at 45 minutes (eNOS) and 2 hours (iNOS) after sildenafil injection. The magnitude of mRNA increase was more pronounced for iNOS than for eNOS. In addition, a significant increase in both iNOS and eNOS protein was detected 24 hours after sildenafil treatment. A selective inhibitor of iNOS, 1400W (10 mg/kg IP given 30 minutes before I-R), abolished sildenafil induced protection (23.7+/-2.8%, P<0.05 versus sildenafil). These data suggest that the induction of NO synthase isoforms is an essential component of the signaling mechanism for sildenafil-induced delayed preconditioning. However, iNOS appears to be the primary isoform that mediates the robust cardioprotection. PMID- 12637372 TI - Efficacy of albumin in critically ill patients. PMID- 12637373 TI - Achieving health for children in public care. PMID- 12637374 TI - Tobacco, coffee, and Parkinson's disease. PMID- 12637375 TI - Massage treatment for back pain. PMID- 12637379 TI - FDA officials argue over safety of new arthritis drug. PMID- 12637380 TI - Gynaecologist faces inquiry after woman nearly dies after abortion. PMID- 12637381 TI - Italy introduces anti-fraud measures. PMID- 12637387 TI - UN report warns that poor water supply continues to pose major health threat. PMID- 12637389 TI - German surgeon under investigation over organ trading. PMID- 12637393 TI - Canadian police charge tobacco companies with smuggling cigarettes. PMID- 12637396 TI - US report warns that lack of health insurance affects overall service provision. PMID- 12637397 TI - Cancer figures for Ireland show increase in lung cancer in women. PMID- 12637398 TI - Australian government bolsters medical scheme for less well off. PMID- 12637399 TI - Bush proposes Medicare reform. PMID- 12637400 TI - Effects of alternative maternal micronutrient supplements on low birth weight in rural Nepal: double blind randomised community trial. AB - OBJECTIVE: To assess the impact on birth size and risk of low birth weight of alternative combinations of micronutrients given to pregnant women. DESIGN: Double blind cluster randomised controlled trial. SETTING: Rural community in south eastern Nepal. PARTICIPANTS: 4926 pregnant women and 4130 live born infants. INTERVENTIONS: 426 communities were randomised to five regimens in which pregnant women received daily supplements of folic acid, folic acid-iron, folic acid-iron-zinc, or multiple micronutrients all given with vitamin A, or vitamin A alone (control). MAIN OUTCOME MEASURES: Birth weight, length, and head and chest circumference assessed within 72 hours of birth. Low birth weight was defined <2500 g. RESULTS: Supplementation with maternal folic acid alone had no effect on birth size. Folic acid-iron increased mean birth weight by 37 g (95% confidence interval -16 g to 90 g) and reduced the percentage of low birthweight babies (<2500 g) from 43% to 34% (16%; relative risk=0.84, 0.72 to 0.99). Folic acid iron-zinc had no effect on birth size compared with controls. Multiple micronutrient supplementation increased birth weight by 64 g (12 g to 115 g) and reduced the percentage of low birthweight babies by 14% (0.86, 0.74 to 0.99). None of the supplement combinations reduced the incidence of preterm births. Folic acid-iron and multiple micronutrients increased head and chest circumference of babies, but not length. CONCLUSIONS: Antenatal folic acid-iron supplements modestly reduce the risk of low birth weight. Multiple micronutrients confer no additional benefit over folic acid-iron in reducing this risk. PMID- 12637401 TI - Impact of DOTS compared with DOTS-plus on multidrug resistant tuberculosis and tuberculosis deaths: decision analysis. AB - OBJECTIVE: This study sought to determine the impact of the World Health Organization's directly observed treatment strategy (DOTS) compared with that of DOTS-plus on tuberculosis deaths, mainly in the developing world. DESIGN: Decision analysis with Monte Carlo simulation of a Markov decision tree. DATA SOURCES: People with smear positive pulmonary tuberculosis. DATA ANALYSIS: Analyses modelled different levels of programme effectiveness of DOTS and DOTS plus, and high (10%) and intermediate (3%) proportions of primary multidrug resistant tuberculosis, while accounting for exogenous reinfection. MAIN OUTCOME MEASURE: The cumulative number of tuberculosis deaths per 100 000 population over 10 years. RESULTS: The model predicted that under DOTS, 276 people would die from tuberculosis (24 multidrug resistant and 252 not multidrug resistant) over 10 years under optimal implementation in an area with 3% primary multidrug resistant tuberculosis. Optimal implementation of DOTS-plus would result in four (1.5%) fewer deaths. If implementation of DOTS-plus were to result in a decrease of just 5% in the effectiveness of DOTS, 16% more people would die with tuberculosis than under DOTS alone. In an area with 10% primary multidrug resistant tuberculosis, 10% fewer deaths would occur under optimal DOTS-plus than under optimal DOTS, but 16% more deaths would occur if implementation of DOTS-plus were to result in a 5% decrease in the effectiveness of DOTS CONCLUSIONS: Under optimal implementation, fewer tuberculosis deaths would occur under DOTS-plus than under DOTS. If, however, implementation of DOTS-plus were associated with even minimal decreases in the effectiveness of treatment, substantially more patients would die than under DOTS. PMID- 12637402 TI - Impact of NICE guidance on laparoscopic surgery for inguinal hernias: analysis of interrupted time series. PMID- 12637403 TI - Bilateral anterior toxic optic neuropathy and the use of infliximab. PMID- 12637404 TI - Quality of care for elderly residents in nursing homes and elderly people living at home: controlled observational study. AB - OBJECTIVES: To assess the quality of care given to elderly people and compare the care given to residents in nursing homes with those living in their own homes. DESIGN: Controlled observational study. SETTING: Primary care, Bristol. SUBJECTS: Elderly individuals (aged > or =65 years) registered with three general practices, of whom 172 were residents in nursing homes (cases) and 526 lived at home (matched controls). MAIN OUTCOME MEASURES: The quality of clinical care given to patients was measured against explicit standards. Quality indicators were derived from national sources and agreed with participating general practitioners. RESULTS: The overall standard of care was inadequate when judged against the quality indicators, irrespective of where patients lived. The overall prescribing of beneficial drugs for some conditions was deficient--for example, only 38% (11/29) (95% confidence interval 20% to 58%) of patients were prescribed beta blockers after myocardial infarction. The proportion of patients with heart disease or diabetes who had had their blood pressure measured in the past two years (heart disease) or past year (diabetes) was lower among those living in nursing homes: for heart disease, 74% (17/23) v 96% (122/127) (adjusted odds ratio 0.18, 0.04 to 0.75); for diabetes, 62% (8/13) v 96% (50/52) (adjusted odds ratio 0.05, 0.01 to 0.38). In terms of potentially harmful prescribing, significantly more patients in nursing homes were prescribed neuroleptic medication (28% (49/172) v 11% (56/526) (3.82, 2.37 to 6.17)) and laxatives (39% (67/172) v 16% (85/526) (2.79, 1.79 to 4.36)). Nursing home residents were less likely to have the appropriate diagnostic Read code linked to their prescribed neuroleptic drug (0.22, 0.07 to 0.71). CONCLUSIONS: The quality of medical care that elderly patients receive in one UK city, particularly those in nursing homes, is inadequate. We suggest that better coordinated care for these patients would avoid the problems of overuse of unnecessary or harmful drugs, underuse of beneficial drugs, and poor monitoring of chronic disease. PMID- 12637405 TI - Management of peripheral arterial disease in primary care. PMID- 12637407 TI - Adrenaline given outside the context of life threatening allergic reactions. PMID- 12637408 TI - Learning and teaching in the clinical environment. PMID- 12637409 TI - Managing patients with inexplicable health problems. PMID- 12637410 TI - Psychological coping and cancer. Study results should not have been dismissed. PMID- 12637411 TI - Erectile dysfunction: NHS should meet current need before preventing future medical needs. PMID- 12637412 TI - Reconfiguring health systems. Potential of new ways of using technology needs to be considered. PMID- 12637413 TI - Effectiveness of trained nurses in preoperative assessment. Unnecessary ritual should not be continued. PMID- 12637414 TI - Channelling new antidepressants to problem patients may be factor in fatal toxicity. PMID- 12637415 TI - Football may influence when patients see a doctor. PMID- 12637416 TI - Recommendations on syncope and driving must be observed. PMID- 12637417 TI - Patient outcomes and population context affect test accuracy. PMID- 12637418 TI - Mind and body split. Philosophy can be detrimental to doctors. PMID- 12637421 TI - Neurolinguistic programming: verbal communication. PMID- 12637422 TI - Flexible training: moving on and up. PMID- 12637423 TI - The characteristics of thoracic insufficiency syndrome associated with fused ribs and congenital scoliosis. AB - 1. Thoracic insufficiency syndrome is the inability of the thorax to support normal respiration or lung growth. 2. The rare condition of fused ribs and congenital scoliosis may result in a three-dimensional thoracic deformity with adverse effects on thoracic growth and function with development of thoracic insufficiency syndrome. 3. The normal thorax is defined by two characteristics: normal, stable volume and the ability to change that volume. Volume depends on the width and depth of the rib cage, and the thoracic spine provides height. The ability to change volume, termed thoracic function, is provided by the diaphragm and the secondary muscles of respiration. 4. On radiographs, the loss of the vertical height of the lung of the concave, restricted hemithorax is defined by the percentage of space available for the lung. 5. Spine rotation causes a windswept thorax, with both restriction of the volume of the convex hemithorax and restriction of the motion of the involved ribs. 6. Constrictive three dimensional deformity of the thorax may cause extrinsic, restrictive lung disease. 7. Progressive thoracic insufficiency syndrome is diagnosed on the basis of clinical signs of respiratory insufficiency, loss of chest wall mobility as demonstrated by the thumb excursion test, worsening indices of three-dimensional thoracic deformity on radiographs and computed tomography scans, or a relative decline in percent predicted vital capacity due to thoracic "failure to thrive," as demonstrated by pulmonary function tests. 8. Treatment of progressive thoracic insufficiency syndrome should provide an acute increase in the thoracic volume with stabilization of any flail chest-wall defects and maintain these improvements as the patient grows, without the need for spine fusion. PMID- 12637424 TI - Growth of the thoracic spine in congenital scoliosis after expansion thoracoplasty. AB - BACKGROUND: Children with congenital thoracic scoliosis associated with fused ribs with a unilateral unsegmented bar adjacent to convex hemivertebrae will invariably have curve progression without treatment. Surgery has been thought to have a negligible growth-inhibition effect on the thoracic spine in such patients because it has been assumed that the concave side of the curve and the unilateral unsegmented bar do not grow, but we are unaware of any conclusive studies regarding this assumption. METHODS: The changes in the length of the concave and convex sides, anterior and posterior vertebral edges, posterior arch, and unilateral unsegmented bars of the thoracic spine were measured in the twenty-one children with congenital scoliosis and fused ribs after expansion thoracoplasty had been carried out with use of a vertical, expandable titanium prosthetic rib. Three of these children had undergone posterior spinal fusion previously. Measurements were made with use of a three-dimensional software program that analyzed baseline and follow-up computed tomography scans. The technique was validated through measurement of the thorax of a small female adult cadaver. RESULTS: The patients without spine fusion had an average age of 3.3 years at the time of the baseline computed tomography scan, and the average duration of follow up was 4.2 years. On the average, these patients showed significant growth (p < 0.0001) of the concave side of the thoracic spine (an increase in length of 7.9 mm/yr, or 7.1%/yr) and the convex side (8.3 mm/yr, or 6.4%/yr) compared with the baseline lengths. There was no significant difference in the increases in length (p = 0.38) between the concave and convex sides. Eleven patients with an unsegmented bar had an average 7.3% increase in the length of the bar (p < 0.0001). In the three children with prior spinal fusion, the increase in length averaged only 4.6 mm/yr (3%/yr) on the concave side of the thoracic spine and 3.7 mm/yr (2.2%/yr) on the convex side; both increases were significant (p < 0.0001). CONCLUSIONS: Longitudinal growth of the thoracic spine in a normal child has been estimated to be 0.6 cm/yr between the ages of five and nine years. After expansion thoracoplasty, growth of the thoracic spine was approximately 8 mm/yr in our series of children with congenital scoliosis and fused ribs. After expansion thoracoplasty, both the concave and the convex side of the thoracic spine and unilateral unsegmented bars appeared to grow in these patients. When a thorax is already foreshortened by congenital scoliosis, control of spine deformity with expansion thoracoplasty allows growth of the thoracic spine, and it is likely that the longer thorax provides additional volume for growth of the underlying lungs with probable clinical benefit. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series [no, or historical, control group]). See Instructions to Authors for a complete description of levels of evidence. PMID- 12637425 TI - Effects of preheating of hip prostheses on the stem-cement interface. AB - BACKGROUND: Debonding of the cement from metal implants has been implicated in the loosening of cemented total hip prostheses. Strengthening of the stem-cement interface has been suggested as a way to prevent loosening of the component. Previously, it was reported that preheating the stem to 44 degrees C reduced the porosity of the cement at the stem-cement interface. The purpose of this study was to determine the effect of stem preheating on the characteristics of the stem cement interface. METHODS: The effects of stem preheating, at temperatures of 37 degrees C, 44 degrees C, and 50 degrees C, on the stem-cement interface were studied in a test model and a preparation that closely simulated the clinical situation. Static interface strength was determined initially and after the stems had been kept in isotonic saline solution at 37 degrees C for two weeks. Fatigue lifetimes were measured, and the nature and extent of porosity at the interface were quantified. RESULTS: Stem preheating had significant effects on the stem cement interface. Stems preheated to 37 degrees C had greater interface shear strength than stems at room temperature both initially (53% greater strength) and after simulated aging (155% greater strength). Fatigue lifetimes were also improved, and there was a >99% decrease in interface porosity. The setting time of the cement decreased 12%, and the maximum temperature at the cement-bone interface increased 6 degrees C. Similar effects were found after preheating to 44 degrees C and 50 degrees C. CONCLUSIONS: Stem preheating had significant effects on the stem-cement interface, with significant improvements in the shear strength and cement porosity of the interface. Also, polymerization temperatures at the cement-bone interface increased. The possible biological effects of these increased interface temperatures at the cement-bone interface require further study. PMID- 12637426 TI - Injury of the femoral nerve associated with acetabular fracture. AB - BACKGROUND: The purpose of the present study was to document the prevalence of, and recovery from, injuries of the femoral nerve associated with displaced acetabular fractures. METHODS: From 1986 to 2001, 726 acetabular fractures were treated with open reduction and internal fixation. Four patients who had an injury of the femoral nerve associated with a displaced acetabular fracture were identified and were followed for a mean of 3.4 years. The nerve injury was iatrogenic in two patients and traumatic in two patients. RESULTS: Clinically detectable quadriceps femoris motor function returned at an average of eighteen weeks (range, four to fifty-two weeks). All patients had satisfactory recovery of nerve function with a return of grade-4 or 5 motor power (the level of motor power needed to allow a normal gait) by an average of ten months (range, three to twenty-four months). Sensory recovery was incomplete but not debilitating. CONCLUSIONS: Preoperative examination of a patient who has an acetabular fracture should include an assessment of femoral nerve function. Regardless of whether the injury is traumatic or iatrogenic in origin, recovery of motor and sensory function without surgical exploration can be expected. LEVEL OF EVIDENCE: Prognostic study, Level II-1 (retrospective study). See Instructions to Authors for a complete description of levels of evidence. PMID- 12637427 TI - Mortality following primary total knee arthroplasty. AB - BACKGROUND: Total joint arthroplasty is one of the most successful orthopaedic surgical procedures. However, it carries a risk of perioperative mortality. The purpose of this study was to determine the mortality rate for patients undergoing primary total knee arthroplasty in a private-practice setting involving one surgeon in a nonteaching institution. METHODS: We analyzed 3048 consecutive primary total knee arthroplasties, performed between July 1976 and December 1996, with respect to mortality data (deaths that occurred intraoperatively, during hospitalization, and within ninety days after surgery) and comorbidities (major cardiovascular disease). RESULTS: The mortality rate was 0.46% (fourteen patients; one death per 217 procedures) within ninety days after primary total knee arthroplasty. CONCLUSIONS: Increasing patient age and the presence of associated cardiovascular comorbidities were identified as risk factors for mortality. This information, we believe, is useful to patients, their families, physicians, and health-care planners in deciding when a total knee arthroplasty is appropriate. LEVEL OF EVIDENCE: Prognostic study, Level II-1 (retrospective study). See Instructions to Authors for a complete description of levels of evidence. PMID- 12637428 TI - Treatment of distal femoral nonunion with internal fixation, cortical allograft struts, and autogenous bone-grafting. AB - BACKGROUND: The purpose of this retrospective study was to analyze the results of treatment of nonunions of the distal part of the femur with internal fixation combined with cortical allograft struts and autogenous bone-grafting. METHODS: Thirteen patients with a distal femoral nonunion were treated with open reduction and internal fixation combined with deep-frozen cortical allograft struts and autogenous iliac bone grafts. The level of nonunion was infra-isthmal in three patients and supracondylar in ten. Ten nonunions were fixed with a blade-plate or condylar buttress plate; two, with an antegrade locked nail; and one, with an antegrade locked nail and a compression plate. Seven unicortical, five bicortical, and one tricortical allograft struts, with an average length of 10 cm, were used. Autogenous iliac bone grafts were inserted into the nonunion site and between the allograft and host femur. The average time from the surgery to the latest follow-up examination was thirty-four months (range, twelve to seventy months). RESULTS: All nonunions united, at an average of five months. The average arc of knee motion improved from 45 degrees preoperatively to 73 degrees postoperatively. According to The Hospital for Special Surgery knee rating system, the average knee score improved from 20 points preoperatively to 71 points at the latest follow-up examination. Three patients had hardware removed after healing of the nonunion, and one patient underwent quadricepsplasty for knee contracture. There were no postoperative infections. Two patients had shortening of the limb of >2 cm. CONCLUSION: Open reduction and internal fixation supplemented with allograft struts and autogenous bone graft is an effective treatment for nonunion of the distal part of the femur. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series [no, or historical, control group]). See Instructions to Authors for a complete description of levels of evidence. PMID- 12637429 TI - Total hip arthroplasty in diastrophic dysplasia. AB - BACKGROUND: Diastrophic dysplasia results in severe disproportionate growth failure, multiple joint deformities, and early osteoarthritis of the hips. Mortality is increased in early childhood, but thereafter life expectancy is normal. Because of severe flexion deformities, resting pain, and diminished movements of the hip joints, total hip arthroplasty is indicated at an early age. The purpose of our study was to evaluate prospectively the midterm results of total hip arthroplasty in a consecutive series of patients with diastrophic dysplasia. METHODS: Between 1982 and 1996, forty-one total hip replacements were performed in twenty-four consecutive patients with diastrophic dysplasia (mean age, forty-one years) at our hospital. The patients were followed prospectively for a minimum of five years with clinical examination, determination of Harris hip scores, and radiographs. Twenty-two patients (thirty-eight hips) were examined clinically and radiographically at the time of follow-up, and the remaining two patients (three hips) were contacted only by telephone. The mean duration of follow-up was 7.8 years. RESULTS: The mean Harris hip score increased from 44 points (range, 25 to 66 points) before the operation to 70 points (range, 37 to 89 points) at the final follow-up examination (p < 0.001). Ten complications (24%) were recorded. Five (12%) of the forty-one hips required revision because of aseptic loosening of the acetabular component at a mean of 9.4 years after the primary operation. No revisions were due to aseptic failure of the femoral component. CONCLUSIONS: Implant survival was good and the Harris hip scores increased significantly after total hip arthroplasty in patients with diastrophic dysplasia. However, shortening femoral osteotomy and transposition of the greater trochanter, adductor and flexor tenotomies, and modification of the femoral stem were frequently needed. Total hip arthroplasty is recommended for patients with diastrophic dysplasia and severe degeneration of the hip joints, even for those who are relatively young. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series [no, or historical, control group]). See Instructions to Authors for a complete description of levels of evidence. PMID- 12637430 TI - Dislocation of rotating hinge total knee prostheses. A biomechanical analysis. AB - BACKGROUND: Symptomatic instability and implant dislocation are occasionally encountered in patients with a rotating hinge total knee prosthesis. A biomechanical study of rotating hinge total knee implants was performed to determine the association between the design (length and taper) of the central rotational stem and the stability of the implant. METHODS: The stem lengths and tapers of knee implants made by seven manufacturers were measured. The tilting laxity of each design was tested by measuring the degree of tilting of the central rotational stem within the tibial housing that occurred with increasing amounts of distraction. The maximum amount of distraction that was possible before the stem dislocated was determined for each design. RESULTS: Implant designs with a short and/or markedly tapered central rotational stem had the greatest tilting, laxity, and instability of that stem. The Howmedica, Techmedica, Intermedics/Sulzer Medica, and Wright Medical Technology/ Dow Corning Wright designs required > or = 39 mm of distraction before they dislocated. The Biomet knee implant required 33 or 44 mm of distraction to dislocate, depending on the thickness of the polyethylene tray that was utilized. The S-ROM knee required only 26 mm of distraction before dislocation occurred. CONCLUSIONS: The measurements confirmed that the shorter the stem and the greater its taper, the greater the instability and laxity at any given amount of joint distraction. PMID- 12637431 TI - Pedicle subtraction osteotomy for the treatment of fixed sagittal imbalance. AB - BACKGROUND: Fixed sagittal imbalance (a syndrome in which the patient is only able to stand with the weight-bearing line in front of the sacrum) has many etiologies. The most commonly reported technique for correction is the Smith Petersen osteotomy. Few reports on pedicle subtraction procedures (resection of the posterior elements, pedicles, and vertebral body through a posterior approach) are available in the peer-reviewed literature. We are aware of no report involving a substantial number of patients with coexistent scoliosis who underwent pedicle/vertebral body subtraction for the treatment of fixed sagittal imbalance. METHODS: Twenty-seven consecutive patients in whom sagittal imbalance was treated with lumbar pedicle subtraction osteotomy at one institution were analyzed. Radiographic analysis included assessment of thoracic kyphosis, lumbar lordosis, lordosis through the pedicle subtraction osteotomy site, and the C7 sagittal plumb line. Outcomes analysis was performed with use of a before-and after pain scale, items from the Oswestry questionnaire, and the Scoliosis Research Society (SRS) questionnaire after a minimum duration of follow-up of two years. Complications and radiographic findings were also analyzed for the entire group. RESULTS: Overall, the average increase in lordosis was 34.1 degrees and the average improvement in the sagittal plumb line was 13.5 cm. One patient had development of a lumbar pseudarthrosis through the area of pedicle subtraction osteotomy, and six patients had development of a thoracic pseudarthrosis. Two patients had development of increased kyphosis at L5/S1, caudad to the fusion, resulting in some loss of sagittal correction. There were significant improvements in the overall Oswestry score (p < 0.0001) and the pain-scale score (p = 0.0002). Most patients reported improvement in terms of pain and self-image as well as overall satisfaction with the procedure. CONCLUSIONS: Pedicle subtraction osteotomy is a useful procedure for patients with fixed sagittal imbalance. A worse clinical result is associated with increasing patient comorbidities, pseudarthrosis in the thoracic spine, and subsequent breakdown caudad to the fusion. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series [no, or historical, control group]). See Instructions to Authors for a complete description of levels of evidence. PMID- 12637432 TI - Effect of terminal sterilization with gas plasma or gamma radiation on wear of polyethylene liners. AB - BACKGROUND: Although terminal sterilization with gamma radiation in air promotes cross-linking, which improves wear resistance, it also results in free radicals, which can oxidize and degrade the mechanical properties of polyethylene liners used for total hip arthroplasty. For this reason, non-cross-linked polyethylene components have also been sterilized with chemical surface treatments, such as gas plasma. In this study, we tested the hypothesis that conventional polyethylene liners cross-linked by sterilization with gamma radiation in air had better in vivo wear performance than non-cross-linked liners sterilized with gas plasma. METHODS: We retrospectively reviewed the wear rates in a series of hips treated with a Duraloc 100 cup, a 28-mm femoral head, and an Enduron liner that had been sterilized with either gamma radiation (sixty-one hips followed for a mean of 5.2 years) or gas plasma (sixty-three hips followed for a mean of 3.9 years). The irradiated liners had been stored with access to ambient oxygen for an average of 1.0 year (range, 0.05 to 5.72 years) prior to implantation. Multiple linear regression was used to assess the effect of the sterilization method on the wear rate while accounting for the possible influence of other factors, including liner geometry, femoral head material, patient gender, cup abduction angle, and age at surgery. RESULTS: The polyethylene liners that had been sterilized with gamma radiation in air had a significantly lower wear rate than did the gas-plasma-sterilized liners (0.097 compared with 0.19 mm/yr, p < 0.001). The sterilization method (p < 0.001) and age at surgery (p = 0.001) were the only factors that we analyzed that correlated with the wear rate. CONCLUSIONS: The in vivo wear of conventional polyethylene liners that had been sterilized with gamma radiation in air was, on the average, 50% less than that of non-cross-linked liners sterilized with gas plasma. LEVEL OF EVIDENCE: Therapeutic study, Level III-2 (retrospective cohort study). See Instructions to Authors for a complete description of levels of evidence. PMID- 12637433 TI - Tibial osteotomy for the treatment of varus gonarthrosis. Survival and failure analysis to twenty-two years. AB - BACKGROUND: The purpose of this retrospective study was to assess the long-term results after the treatment of medial compartment gonarthrosis with a proximal tibial osteotomy to determine whether various clinical and radiographic factors relate to the outcome. METHODS: From 1972 to 1990, seventy-six valgus-producing high tibial osteotomies were performed in sixty-six patients with medial compartment gonarthrosis. Stabilization was achieved with use of a plate on the lateral side of the proximal part of the tibia and a figure-of-eight wire on the medial side. The mean age of the patients at the time of surgery was sixty-nine years, and the mean duration of follow-up was 10.8 years. The factors that were analyzed included postoperative valgus alignment; the age, gender, and weight of the patient; preoperative Ahlback radiographic grade; adverse events; Workers' Compensation status; and public liability. Survivorship outcome end points were conversion to arthroplasty, a Hospital for Special Surgery knee score of <70 points, and patient dissatisfaction with the procedure. RESULTS: Ten-year survival, according to the life-table method, was 74%, 70%, and 65% with conversion to arthroplasty, a Hospital for Special Surgery knee score of <70 points, and patient dissatisfaction, respectively, as the end points. Radiographic valgus alignment that ranged between 8 degrees and 16 degrees at one year after the osteotomy had the most significant positive effect on survivorship for all end points (p < 0.01) compared with the other parameters. Complications occurred in sixteen (21%) of the seventy-six procedures, and they had a significant effect on survival, with all three end points (p < 0.05). CONCLUSIONS: We found that survival at ten years was 90% (95% confidence interval, 80% to 99%) when the radiographic valgus angle at one year was between 8 degrees and 16 degrees with arthroplasty as the end point. We believe that there is a role for tibial osteotomy, as an alternative to total knee arthroplasty, in patients who are less than sixty years old. LEVEL OF EVIDENCE: Therapeutic study, Level III-2 (retrospective cohort study). See Instructions to Authors for a complete description of levels of evidence. PMID- 12637434 TI - Microcirculation associated with degenerative rotator cuff lesions. In vivo assessment with orthogonal polarization spectral imaging during arthroscopy of the shoulder. AB - BACKGROUND: Diminished vascular supply is associated with degenerative rotator cuff lesions. Orthogonal polarization spectral imaging allows noninvasive assessment of microcirculation without application of fluorescent contrast medium. The aim of our study was to visualize and quantify in vivo the microcirculation of the rotator cuff during arthroscopic surgery and to compare the results with the number of microvessels identified in vitro by immunostaining of biopsy specimens taken from the scanned areas. METHODS: Eleven patients with clinical signs of a degenerative rotator cuff lesion were studied. Prior to arthroscopic subacromial decompression, the superficial part of the supraspinatus tendon at the edge of the lesion as well as the unaffected tendon insertion was examined. Microvascular parameters established for the description of tissue perfusion with use of conventional intravital fluorescence microscopy (functional capillary density and capillary diameter) were assessed in vivo. Biopsy specimens were taken from the scanned areas, and the microvessels were localized by immunostaining for the endothelial surface marker CD31. RESULTS: In the region of the unaffected tendon insertion, the mean baseline functional capillary density (and standard deviation) was 106 +/- 13 cm/cm(2) and the mean capillary diameter was 10 +/- 0.7 microm. In contrast, at the edge of the lesion, the functional capillary density was significantly reduced to 20 +/- 14 cm/cm(2), whereas the diameter of the vessels that were present did not differ. The total number of vessels stained in vitro was also significantly reduced at the edge of the lesion compared with the number of vessels in the tendon insertion zone. CONCLUSIONS: Quantitative in vivo analysis of human microcirculation during arthroscopy demonstrated that the functional capillary density at the edge of a degenerative rotator cuff lesion was significantly reduced compared with that in the control tissue. The capacity to assess microcirculatory flow in vivo may help to identify specific repair strategies based on knowledge of individual perfusion patterns. PMID- 12637435 TI - Femoral anteversion in children with cerebral palsy. Assessment with two and three-dimensional computed tomography scans. AB - BACKGROUND: Assessment of femoral anteversion in children with cerebral palsy with two or three-dimensional computed tomography scans may be limited by both positional and anatomic variables. Three-dimensional computed tomography techniques are considered to be more accurate than two-dimensional imaging when the femur is not optimally positioned in the gantry or when the neck-shaft angle is increased. METHODS: Computed tomography scanning was performed on a series of nine model femora with anteversion ranging from 20 degrees to 60 degrees and neck shaft angles ranging from 120 degrees to 160 degrees. Each femoral model was scanned in two holding devices, the first of which held the femur in optimal alignment (normal model) and the second of which held the femur in flexion, adduction, and internal rotation (cerebral palsy model) relative to the gantry. Femoral anteversion was calculated for each model from two and three-dimensional computed tomography scans by four examiners on two separate occasions. The intraobserver and interobserver reliability, the accuracy, and the effect of increasing the neck-shaft angle on the accuracy of the measurements made on the two and three-dimensional scans of the normal and cerebral palsy models were then examined. RESULTS: The mean differences in the measurements of femoral anteversion made by the same examiner (intraobserver reliability) were <2 degrees for the two-dimensional scans of the normal and cerebral palsy models and the three-dimensional scans of the normal models, and the mean difference was <4 degrees for the three-dimensional scans of the cerebral palsy models. The mean differences among examiners (interobserver reliability) were <3 degrees for the two-dimensional scans of the normal and cerebral palsy models and the three dimensional scans of the normal models, and the mean difference was <6 degrees for the three-dimensional scans of the cerebral palsy models. The accuracy of the assessments of femoral anteversion of the normally aligned models was comparable between the two and three-dimensional scans. However, the three-dimensional assessment was significantly more accurate than the two-dimensional assessment for measurement of anteversion of the cerebral palsy models (p = 0.003). Accuracy within 5 degrees was comparable between the two and three-dimensional scans for measurement of the normally aligned models, with 86% of the two-dimensional measurements and 78% of the three-dimensional measurements falling within 5 degrees of the actual measurements. However, the accuracy within 5 degrees was significantly compromised when the models were placed in cerebral palsy alignment. Only 3% of the two-dimensional measurements and 14% of the three dimensional measurements fell within 5 degrees of the actual measurements, with three-dimensional assessment being significantly better than two-dimensional assessment (p = 0.006). Increasing the neck-shaft angle did not significantly compromise the accuracy of measurement of femoral anteversion with either the two dimensional or the three-dimensional technique (p > 0.05 for all comparisons). CONCLUSIONS: When adequate alignment of the femur in the computed tomography scanner was possible, a simple two-dimensional technique exhibited excellent intraobserver and interobserver reliability and clinically acceptable accuracy within the relevant ranges of anatomic variability tested (neck-shaft angles of 120 degrees to 160 degrees and femoral anteversion of 20 degrees to 60 degrees). When optimal alignment of the femur in the scanner was not possible, neither two dimensional nor three-dimensional techniques exhibited clinically acceptable accuracy for the measurement of femoral anteversion. PMID- 12637436 TI - The AGC all-polyethylene tibial component: a ten-year clinical evaluation. AB - BACKGROUND: While high success rates have been achieved in association with other all-polyethylene tibial components, an alarming number of failures have occurred at our institution in association with the use of an all-polyethylene version of the AGC tibial component. The purpose of the present study was to describe the survival of the AGC all-polyethylene tibial component. METHODS: Five hundred and thirty-six AGC all-polyethylene tibial components were implanted in 405 patients and were followed over a ten-year period. The average age of the patients at the time of surgery was 70.3 years, the average weight was 78 kg, and the most common diagnosis was osteoarthritis (prevalence, 92.9%). A clinical and radiographic analysis was performed, Knee Society knee and function scores were determined, and Kaplan-Meier survivorship analysis was conducted. Failure was defined as aseptic loosening as evidenced by progressive radiolucent lines and/or revision due to aseptic loosening or collapse. RESULTS: A high rate of failure was noted in the early postoperative period, with a survival rate of 90.04% (95% confidence interval, 87.35% to 92.72%) after three years. At ten years, the survival rate was 68.11% (95% confidence interval, 57.57% to 78.65%). Fifty-eight (73.4%) of seventy-nine failures occurred in association with loosening or collapse of the bone beneath the medial tibial plateau. CONCLUSION: While some all-polyethylene tibial designs have been successful, the low success rate among knees treated with the AGC all-polyethylene tibial component suggests that the results associated with all-polyethylene tibial components are design-sensitive. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series [no, or historical, control group]). See Instructions to Authors for a complete description of levels of evidence. PMID- 12637437 TI - Reproducibility of the radiographic metatarsophalangeal angle in hallux surgery. AB - BACKGROUND: Five different methods have been described to define the longitudinal axis of the first metatarsal for radiographic measurements of the first metatarsophalangeal angle. None of these methods has been validated for both preoperative and postoperative assessment of patients undergoing hallux valgus surgery. Previous studies have demonstrated conflicting results regarding the measurement accuracy of these methods. METHODS: To evaluate the measurement accuracy of these five methods, we calculated the intraobserver and interobserver coefficients of repeatability for all five methods with use of twenty preoperative and twenty postoperative standardized plain dorsoplantar weight bearing radiographs of patients undergoing chevron distal osteotomy. RESULTS: The preoperative assessment of the metatarsophalangeal angle revealed small differences among the five methods. The intraobserver coefficient of repeatability ranged from 2.10 degrees to 3.34 degrees, and the interobserver coefficient ranged from 2.17 degrees to 3.44 degrees. The postoperative assessment demonstrated substantial differences between methods in which the diaphysis of the first metatarsal is used as a reference (intraobserver coefficient, 5.06 degrees to 7.23 degrees; interobserver coefficient, 5.29 degrees to 8.19 degrees) and methods in which there is one reference point in the metatarsal head and one reference point in the base of the first metatarsal (intraobserver coefficient, 1.88 degrees to 2.67 degrees; interobserver coefficient, 1.86 degrees to 2.34 degrees). CONCLUSIONS: For the assessment of patients undergoing a distal metatarsal osteotomy, we cannot recommend methods in which the metatarsal shaft is used as a reference for the axis of the first metatarsal. Such methods had poor measurement accuracy, especially postoperatively. Methods with reference points distal and proximal to any possible osteotomy site had much better measurement reproducibility. We recommend the method described by Miller in 1974, in which a line is drawn from the center of the first metatarsal head through the center of the base of the first metatarsal, as it was the most precise method and was least biased by postoperative effects. PMID- 12637438 TI - The natural history of symptomatic osteonecrosis in adults with sickle-cell disease. AB - BACKGROUND: Adult patients with sickle-cell disease are at risk for the development of osteonecrosis of the hip. However, there is little information in the literature about the rate of progression of osteonecrosis once symptoms begin. The purpose of this study was to evaluate the natural history of the symptomatic hip in adult patients with osteonecrosis and sickle-cell disease. METHODS: Ninety-two symptomatic hips in sixty-four consecutive adult patients with sickle-cell disease were initially evaluated between 1980 and 1987. Sixty symptomatic hips had radiographic evidence of osteonecrosis at the initial evaluation: forty-three were classified as stage II; two, as stage III; and fifteen, as stage IV, according to the system of Steinberg et al. The other thirty-two hips had lesions (stage I) that were evident only on magnetic resonance imaging. All patients were evaluated after a mean duration of follow-up of seventeen years. RESULTS: Of the seventy-five hips without collapse of the femoral head at the initial evaluation, sixty-five demonstrated collapse within five years after the diagnosis. The average time between the diagnosis and collapse was forty-two months for stage-I hips and thirty months for stage-II hips. At the most recent follow-up examination, ninety hips had had collapse of the femoral head and eighty-eight of the ninety-two hips had had surgery because of intractable pain. CONCLUSIONS: Symptomatic osteonecrosis of the hip in sickle cell disease has a high likelihood of leading to femoral head collapse, necessitating surgical intervention. When osteonecrosis develops, the deterioration is rapid and, in most patients, operative intervention is necessary because of intractable pain. LEVEL OF EVIDENCE: Prognostic study, Level II-1 (retrospective study). See Instructions to Authors for a complete description of levels of evidence. PMID- 12637439 TI - Validation of two and three-dimensional radiographic techniques for measuring polyethylene wear after total hip arthroplasty. AB - BACKGROUND: Three-dimensional radiographic techniques have been developed to estimate in vivo polyethylene wear of total hip replacements. We are not aware of any published study examining the accuracy of these in vivo methods. Our objective was to validate two radiographic techniques by comparing their results with those obtained directly from retrieved specimens. METHODS: A coordinate measuring machine was used to probe the interior bearing surface of seventeen acetabular liners that had been retrieved at revision surgery. Solid models were created to determine volumetric polyethylene loss and linear depth and direction of wear. Two in vivo radiographic techniques (PolyWare and the Martell Hip Analysis Suite) were used to calculate two-dimensional linear, three-dimensional linear, and volumetric wear. The radiographic analysis was done independently, and the results were compared with the known values of polyethylene wear derived with use of the coordinate measuring machine. RESULTS: Correlation coefficients comparing the values for two-dimensional and three-dimensional linear wear and volumetric wear derived with the PolyWare radiographic technique with the values derived with the coordinate measuring machine were r(2) = 0.78, r(2) = 0.75, and r(2) = 0.91, respectively (all p < 0.001), and the correlation coefficients comparing the values derived with the Martell Hip Analysis Suite radiographic technique with those derived with the coordinate measuring machine were r(2) = 0.80, r(2) = 0.84, and r(2) = 0.91, respectively (p < 0.001). The average absolute difference between the radiographic estimates and the results derived with the coordinate measuring machine was approximately 19% (range, 13% to 24%). CONCLUSIONS: There was good agreement between the wear estimates made with both in vivo techniques and the measurements of the retrieved polyethylene liners made with the coordinate measuring machine. Two-dimensional wear analysis (based on anteroposterior radiographs) accounted for most of the polyethylene wear, while one technique of three-dimensional wear analysis (PolyWare) demonstrated some additional wear in the lateral plane. PMID- 12637440 TI - Computed tomographic assessment of fractures of the posterior wall of the acetabulum after operative treatment. AB - BACKGROUND: The purpose of this study was to evaluate the results after operative treatment of fractures of the posterior wall of the acetabulum in relationship to the quality of the fracture reduction as assessed by postoperative two dimensional computed tomography. METHODS: The functional results for sixty-seven patients who had open reduction and internal fixation of an unstable fracture of the posterior wall of the acetabulum and the findings of two-dimensional computed tomography performed postoperatively were analyzed. Sixty-one patients were followed for a mean of four years after the injury, and the remaining six patients who had poor early results necessitating reconstructive surgery were followed for less than two years. All patients were evaluated preoperatively and postoperatively with use of three standard plain radiographs (one anteroposterior and two Judet 45 degrees oblique pelvic radiographs) and a two-dimensional computed tomography scan. The functional outcome for the patients was evaluated with use of a modification of the clinical grading system described by Letournel and Judet. The radiographs were graded according to the criteria described by Matta. The two-dimensional computed tomography scans were used to determine fracture gap and offset measurements. RESULTS: The clinical outcome was graded as excellent in thirty-one patients (46%), very good in twenty (30%), good in eight (12%), and poor in eight (12%). The final radiographic results were graded as excellent in fifty-three hips (79%), good in four (6%), fair in three (5%), and poor in seven (10%). There was a strong association between clinical outcome and final radiographic grade. Fracture reductions were graded as anatomic in sixty five and imperfect in two, as determined with use of plain radiography. However, postoperative computed tomography revealed an incongruency (offset) of >2 mm in eleven hips and fracture gaps of > or = 2 mm in fifty-two. Fracture gaps of > or = 10 mm in any dimension or a total gap area of > or = 35 mm(2) were associated with a poor result. The main risk factors for a poor result were a residual fracture gap width of > or = 10 mm and osteonecrosis of the femoral head. CONCLUSIONS: The degree of residual fracture displacement is detected more accurately on postoperative computed tomography scans than on plain radiographs. The accuracy of surgical reduction as assessed on postoperative computed tomography is highly predictive of the clinical outcome. LEVEL OF EVIDENCE: Therapeutic study, Level III-2 (retrospective cohort study). See Instructions to Authors for a complete description of levels of evidence. PMID- 12637441 TI - Outcomes of total hip arthroplasty and contralateral bipolar hemiarthroplasty: a case series. PMID- 12637442 TI - Acute compartment syndrome of the foot following an inversion injury of the ankle with disruption of the anterior tibial artery. A case report. PMID- 12637443 TI - Intracortical chondrosarcoma. A case report. PMID- 12637444 TI - Dissociation of a Morse-taper stemmed tibial component following revision total knee arthroplasty. A case report. PMID- 12637445 TI - Biologic aspects of flexor tendon laceration and repair. PMID- 12637447 TI - Curricular requirements for musculoskeletal medicine in American medical schools. PMID- 12637448 TI - Roles of the medial collateral ligament and the coronoid in elbow stability. PMID- 12637449 TI - Gender and racial differences in rates of total hip replacement. PMID- 12637450 TI - Fat embolism versus fat embolization following total hip arthroplasty. PMID- 12637451 TI - Mechanism of injury may affect outcome after tibial shaft fracture. PMID- 12637452 TI - Subchondral insufficiency fracture of the femoral head and acetabulum: indications for total hip arthroplasty. PMID- 12637453 TI - Educational opportunities for medical students and young orthopaedic surgeons. PMID- 12637454 TI - What's new in foot and ankle surgery. PMID- 12637456 TI - Are anthracycline-taxane regimens the new standard of care in the treatment of metastatic breast cancer? PMID- 12637457 TI - Optimal chemotherapy for women with breast cancer: the plot thickens. PMID- 12637458 TI - Lymph node dissection: is it diagnostic or therapeutic? PMID- 12637460 TI - Improved outcomes from adding sequential Paclitaxel but not from escalating Doxorubicin dose in an adjuvant chemotherapy regimen for patients with node positive primary breast cancer. AB - PURPOSE: This study was designed to determine whether increasing the dose of doxorubicin in or adding paclitaxel to a standard adjuvant chemotherapy regimen for breast cancer patients would prolong time to recurrence and survival. PATIENTS AND METHODS: After surgical treatment, 3,121 women with operable breast cancer and involved lymph nodes were randomly assigned to receive a combination of cyclophosphamide (C), 600 mg/m(2), with one of three doses of doxorubicin (A), 60, 75, or 90 mg/m(2), for four cycles followed by either no further therapy or four cycles of paclitaxel at 175 mg/m(2). Tamoxifen was given to 94% of patients with hormone receptor-positive tumors. RESULTS: There was no evidence of a doxorubicin dose effect. At 5 years, disease-free survival was 69%, 66%, and 67% for patients randomly assigned to 60, 75, and 90 mg/m(2), respectively. The hazard reductions from adding paclitaxel to CA were 17% for recurrence (adjusted Wald chi(2) P =.0023; unadjusted Wilcoxon P =.0011) and 18% for death (adjusted P =.0064; unadjusted P =.0098). At 5 years, the disease-free survival (+/- SE) was 65% (+/- 1) and 70% (+/- 1), and overall survival was 77% (+/- 1) and 80% (+/- 1) after CA alone or CA plus paclitaxel, respectively. The effects of adding paclitaxel were not significantly different in subsets defined by the protocol, but in an unplanned subset analysis, the hazard ratio of CA plus paclitaxel versus CA alone was 0.72 (95% confidence interval, 0.59 to 0.86) for those with estrogen receptor-negative tumors and only 0.91 (95% confidence interval, 0.78 to 1.07) for patients with estrogen receptor-positive tumors, almost all of whom received adjuvant tamoxifen. The additional toxicity from adding four cycles of paclitaxel was generally modest. CONCLUSION: The addition of four cycles of paclitaxel after the completion of a standard course of CA improves the disease free and overall survival of patients with early breast cancer. PMID- 12637459 TI - Docetaxel and doxorubicin compared with doxorubicin and cyclophosphamide as first line chemotherapy for metastatic breast cancer: results of a randomized, multicenter, phase III trial. AB - PURPOSE: This randomized, multicenter, phase III study compared doxorubicin and docetaxel (AT) with doxorubicin and cyclophosphamide (AC) as first-line chemotherapy (CT) in metastatic breast cancer (MBC). PATIENTS AND METHODS: Patients (n = 429) were randomly assigned to receive doxorubicin 50 mg/m(2) plus docetaxel 75 mg/m(2) (n = 214) or doxorubicin 60 mg/m(2) plus cyclophosphamide 600 mg/m(2) (n = 215) on day 1, every 3 weeks for up to eight cycles. RESULTS: Time to progression (TTP; primary end point) and time to treatment failure (TTF) were significantly longer with AT than AC (median TTP, 37.3 v 31.9 weeks; log rank P =.014; median TTF, 25.6 v 23.7 weeks; log-rank P =.048). The overall response rate (ORR) was significantly greater for patients taking AT (59%, with 10% complete response [CR], 49% partial response [PR]) than for those taking AC (47%, with 7% CR, 39% PR) (P =.009). The ORR was also higher with AT in patients with visceral involvement (58% v 41%; liver, 62% v 42%; lung, 58% v 35%), three or more organs involved (59% v 40%), or prior adjuvant CT (53% v 41%). Overall survival (OS) was comparable in both arms. Grade 3/4 neutropenia was frequent in both groups, although febrile neutropenia and infections were more frequent for patients taking AT (respectively, 33% v 10%, P <.001; 8% v 2%, P =.01). Severe nonhematologic toxicity was infrequent in both groups, including grade 3/4 cardiac events (AT, 3%; AC, 4%). CONCLUSION: AT significantly improves TTP and ORR compared with AC in patients with MBC, but there is no difference in OS. AT represents a valid option for the treatment of MBC. PMID- 12637461 TI - Randomized trial of tamoxifen versus tamoxifen plus aminoglutethimide as adjuvant treatment in postmenopausal breast cancer patients with hormone receptor-positive disease: Austrian breast and colorectal cancer study group trial 6. AB - PURPOSE: To determine whether the addition of aminoglutethimide to tamoxifen is able to improve the outcome in postmenopausal patients with hormone receptor positive, early-stage breast cancer. PATIENTS AND METHODS: A total of 2,021 postmenopausal women were randomly assigned to receive either tamoxifen for 5 years alone or tamoxifen in combination with aminoglutethimide (500 mg/d) for the first 2 years of treatment. Tamoxifen was administered at 40 mg/d for the first 2 years and at 20 mg/d for 3 years. RESULTS: All randomized and eligible patients were included in the analysis according to the intention-to-treat principle. After a median follow-up of 5.3 years, the 5-year disease-free survival in the aminoglutethimide plus tamoxifen group was 83.6% versus 83.7% in the monotherapy group (P =.89). The corresponding data for overall survival at 5 years were 91.4% and 91.2%, respectively (P =.74). More patients failed to complete combination treatment (13.7%) because of side effects as compared to tamoxifen alone (5.2%; P =.0001). CONCLUSION: Aminoglutethimide given for 2 years in addition to tamoxifen for 5 years does not improve the prognosis of postmenopausal patients with receptor-positive, lymph node-negative or lymph node-positive breast cancer. PMID- 12637462 TI - Randomized controlled trial comparing oral doxifluridine plus oral cyclophosphamide with doxifluridine alone in women with node-positive breast cancer after primary surgery. AB - PURPOSE: We compared the therapeutic usefulness of doxifluridine (5'-DFUR) alone and a combination of 5'-DFUR plus cyclophosphamide (CPM), both of which are considered effective against advanced and recurrent breast cancer, to determine which treatment is more beneficial as postoperative adjuvant chemotherapy. PATIENTS AND METHODS: A total of 1,131 women with node-positive primary breast cancer were randomly assigned after primary surgery to receive 5'-DFUR alone or 5'-DFUR plus CPM. All patients initially received 5'-DFUR in an oral dose of 1,200 mg/d for 4 weeks, starting 4 weeks after surgery. Chemotherapy was then not given for 2 weeks. Patients in the 5'-DFUR group subsequently received five 4 week cycles of treatment consisting of oral 5'-DFUR (1,200 mg/d) for the first 2 weeks and no chemotherapy for the next 2 weeks. Those assigned to the 5'-DFUR plus CPM group also received oral CPM 100 mg/d for the first 2 weeks and no chemotherapy for the next 2 weeks. Women 50 years or older concurrently received 20 mg/d of tamoxifen for 2 years in both groups. RESULTS: Of the 1,088 eligible women, 546 were assigned to receive 5'-DFUR alone and 542 were assigned to receive 5'-DFUR plus CPM. Overall disease-free survival was significantly better in women who received 5'-DFUR plus CPM than in those who received 5'-DFUR alone (log-rank test, P =.021). Toxic effects occurred in 20.0% of patients (109 of 546) in the 5'-DFUR group and 32.3% of patients (175 of 542) in the 5'-DFUR plus CPM group (chi(2) test, P <.001). CONCLUSION: Combination therapy with 5'-DFUR plus CPM is more effective in preventing recurrence than 5'-DFUR alone. PMID- 12637463 TI - Multicenter phase II study of oral bexarotene for patients with metastatic breast cancer. AB - PURPOSE: Bexarotene is a retinoid X receptor-selective retinoid that has preclinical antitumor activity in breast cancer. We evaluated the efficacy and safety of oral bexarotene in the treatment of patients with metastatic breast cancer. PATIENTS AND METHODS: The following three groups of patients were treated: hormone-refractory, chemotherapy-refractory, and tamoxifen-resistant patients. Patients in the first two groups were treated with bexarotene alone, whereas the tamoxifen-resistant patients received both tamoxifen and bexarotene. Patients in all groups were randomly assigned to receive bexarotene at either 200 or 500 mg/m(2)/d. RESULTS: One hundred forty-eight patients were randomized; 145 patients were treated. Of 48 hormone-refractory patients, there were two partial responses (6%) and 10 patients with stable disease lasting more than 6 months; of 47 chemotherapy-refractory patients, there were two partial responses (6%) and five patients with stable disease; and of 51 tamoxifen-resistant patients, there was one partial response (3%) and 11 patients with stable disease. All partial responses occurred at the 200-mg/m(2)/d dose. The projected median time to progression across all of the arms was 8 to 10 weeks. There were no drug-related deaths, and only two patients had drug-related serious adverse events. The most common drug-related adverse events were hypertriglyceridemia (84%), dry skin (34%), asthenia (30%), and headache (27%). There were no cases of pancreatitis. CONCLUSION: The efficacy of bexarotene in patients with refractory metastatic breast cancer is limited. However, it is an oral agent with minimal toxicity and a unique mechanism of action, which produced clinical benefit in approximately 20% of patients. Future efforts should define populations likely to benefit from this agent. PMID- 12637464 TI - Phase II, randomized, double-blind study of two dose levels of arzoxifene in patients with locally advanced or metastatic breast cancer. AB - PURPOSE: To select a daily dose of arzoxifene (LY353381), a selective estrogen receptor modulator, for use in future studies in women with locally advanced or metastatic breast cancer who are either potentially tamoxifen sensitive (TS) or tamoxifen refractory (TR). PATIENTS AND METHODS: This trial was a randomized, double-blind, phase II study of arzoxifene 20 mg (n = 55) and 50 mg (n = 57) in women with advanced or metastatic breast cancer. Patients were randomly assigned to balance for number of metastatic disease sites, prior tamoxifen therapy, and estrogen receptor status. The primary end point was tumor response rate (RR). Secondary end points included clinical benefit rate (CBR), time to progression (TTP), and toxicity. RESULTS: Forty-nine patients were TS and 63 were TR. According to independent review, among TS patients, RR was higher in the 20-mg arm than the 50-mg arm (26.1% v 8.0%), with a longer TTP (8.3 v 3.2 months; P >.05). Among the TR patients, response rate was the same in the 20-mg and 50-mg arms (10.3%) with similar TTP (2.7 and 2.8 months, respectively; P >.05). CBR was higher in the 20-mg arm than in the 50-mg arm among TS patients (39.1% v 20.0%) and TR patients (13.8% v 10.3%). Arzoxifene was well tolerated. Dose-dependent toxicity was not demonstrated. There were no deaths during study. CONCLUSION: Arzoxifene is effective in the treatment of TS and TR patients with advanced or metastatic breast cancer at the 20-mg and 50-mg dose levels. Toxicities are minimal, and the therapy is tolerated. The 20-mg dose seems to be at least as effective as the 50-mg dose. Accordingly, arzoxifene 20 mg/d was selected for further study in patients with breast cancer. PMID- 12637465 TI - Immunotherapy of advanced breast cancer with a heterophilic ganglioside (NeuGcGM3) cancer vaccine. AB - PURPOSE: A heterophilic ganglioside cancer vaccine was developed by combining NeuGcGM3 with the outer membrane protein complex of Neisseria meningitidis to form very small size proteoliposomes (VSSP). A phase I clinical trial was performed to determine safety and immunogenicity of this vaccine. PATIENTS AND METHODS: Stage III to IV breast cancer patients received up to 15 (200 micro g) doses of the vaccine by intramuscular injection. The first five doses (induction phase) were given at 2-week intervals, with the remaining treatment (maintenance) administered on a monthly basis. RESULTS: Twenty-one patients, 11 of whom had metastatic disease, were included. Main toxicities included erythema and induration at the injection site, sometimes associated with mild pain, and low grade fever (World Health Organization grades 1 and 2). All treated patients who completed the induction phase developed anti-NeuGcGM3 antibody titers between 1:1,280 and 1:164,000 immunoglobulin G (IgG), and 1:640 and 1:164,000 IgM. Noteworthy specific IgA antibodies were induced by vaccination in all stage III patients and in three stage IV patients. Serum antibody levels were higher in the stage III patients, with the larger increases observed after week 32. The antiganglioside IgG subclasses were mainly IgG1 and IgG3. Hyperimmune sera increased complement-mediated cytotoxicity versus P3X63 myeloma cells and a marked IgG differential reactivity against human mammary ductal carcinoma samples. CONCLUSION: NeuGcGM3/VSSP/Montanide ISA 51 is an unusual immunogenic ganglioside vaccine and also seems to be safe in this small trial. Immunologic surrogates of activity indicate that this reagent warrants further investigation. PMID- 12637466 TI - Invasion factors uPA/PAI-1 and HER2 status provide independent and complementary information on patient outcome in node-negative breast cancer. AB - PURPOSE: The independent clinical relevance of invasion factors urokinase-type plasminogen activator (uPA)/PAI-1 and HER2 status was evaluated in lymph node negative breast cancer patients (N = 118) without adjuvant systemic therapy after long-term follow-up of more than 10 years (median, 126 months). PATIENTS AND METHODS: Levels of uPA and its inhibitor PAI-1 were prospectively measured by enzyme-linked immunosorbent assay in primary tumor tissue extracts. HER2 gene amplification (HER2_AMP) was evaluated by fluorescence in situ hybridization (FISH; Ventana Medical Systems HER-2/neu probe; Tucson, AZ), and HER2 protein overexpression (HER2_EXP) was evaluated by immunohistochemistry (IHC; Oncogene Science antibody Ab-3; Cambridge, MA) on parallel-cut formalin-fixed paraffin embedded tissue sections. RESULTS: uPA/PAI-1 was high (either one or both factors were high) in 44% of the tumors. HER2_AMP was detected by FISH in 33% of the patients, and HER2_EXP was found by IHC in 44% of the patients. In a multivariate analysis of established and tumor-biologic prognostic factors, uPA/PAI-1 was the only independent prognostic factor for disease-free survival ([DFS]; P <.001; relative risk [RR], 8.3; 95% confidence interval [CI], 3.4 to 20.4). Although HER2_AMP and HER2_EXP did not reach significance for DFS, they were significant for overall survival (OS), even in multivariate analysis (HER2_AMP: P =.004; RR, 3.7; 95% CI, 1.5 to 9.2; HER2_EXP: P =.009; RR, 3.4; 95% CI, 1.4 to 8.7). CONCLUSION: After long-term follow-up, uPA/PAI-1 levels in primary tumor tissue reliably and strongly indicate an aggressive course of disease in lymph node negative breast cancer independent of HER2 status. The particular prognostic effect of HER2 status on OS may reflect its ability to predict resistance to systemic therapy. PMID- 12637467 TI - Effect of number of lymph nodes sampled on outcome in patients with stage I non small-cell lung cancer. AB - PURPOSE: We postulate that surgical sampling and pathologic evaluation of lymph nodes of surgical specimens from patients with stage I non-small-cell lung cancer (NSCLC) can have an effect on the time to recurrence and survival of these patients. PATIENTS AND METHODS: We analyzed data on 442 patients with stage I NSCLC who were treated with surgical resection and some form of lymph node sampling. Associations between total lymph nodes sampled and disease-free survival (DFS) and overall survival (OS) were investigated. The effect of total lymph node stations sampled and the surgical techniques (random lymph node sampling, systematic sampling [SS], or complete mediastinal lymph node dissection [MLND]) on DFS and OS was also studied. Complete MLND and SS were defined as dissection or sampling of levels 4, 7, and 10 for right-sided lesions and levels 5 or 6 and 7 for left-sided lesions. RESULTS: Patients were divided into quartiles on the basis of total number of lymph nodes sampled. Improved DFS and OS were associated with greater number of lymph nodes sampled. SS and MLND were associated with improved survival compared with random lymph node sampling. The total number of lymph nodes sampled maintained strong significance in the multivariate analysis. CONCLUSION: These results indicate that examining a greater number of lymph nodes in patients with stage I NSCLC treated with resection increases the likelihood of proper staging and affects patient outcome. Such information is important not only for therapy and prognosis of individuals but also for identifying those who may benefit from adjuvant therapy. PMID- 12637468 TI - Human kallikrein 6 (hK6): a new potential serum biomarker for diagnosis and prognosis of ovarian carcinoma. AB - PURPOSE: The discovery of new ovarian cancer biomarkers that are suitable for early disease diagnosis and prognosis may ultimately lead to improved patient management and outcomes. PATIENTS AND METHODS: We measured, by immunoassay, human kallikrein 6 (hK6) concentration in serum of 97 apparently healthy women, 141 women with benign abdominal diseases, and 146 women with histologically proven primary ovarian carcinoma. We then calculated the diagnostic sensitivity and specificity of this test and examined the association of serum hK6 concentration with various clinicopathologic variables and patient survival. RESULTS: Serum hK6 concentration between normal and benign disease patients was not different (mean, 2.9 and 3.1 micro g/L, respectively). However, hK6 in presurgical serum of ovarian cancer patients was highly elevated (mean, 6.8 micro g/L; P <.001). Serum hK6 decreased after surgery (to a mean of 3.9 micro g/L) in 68% of patients. The diagnostic sensitivity of serum hK6 at 90% and 95% specificity is 52% and 47%, respectively, in the whole patient population. For early stage disease (stage I or II), sensitivity is approximately 21% to 26%. When combined with CA-125, at 90% specificity, sensitivity increases to 72% (for all patients) and to 42% in stage I or II disease. Serum hK6 concentration correlates moderately with CA-125 and is higher in patients with late-stage, higher-grade disease and in patients with serous histotype. Preoperative serum hK6 concentration is a powerful predictor of disease-free and overall survival in both univariate and multivariate analyses. CONCLUSIONS: Serum hK6 concentration seems to be a new biomarker for ovarian carcinoma and may have value for disease diagnosis and prognosis. PMID- 12637469 TI - Treatment of primary CNS lymphoma with methotrexate and deferred radiotherapy: a report of NABTT 96-07. AB - PURPOSE: A multicenter, phase II study of single-agent, intravenous methotrexate in newly diagnosed non-AIDS-related primary CNS lymphoma was conducted in the New Approaches to Brain Tumor Therapy (NABTT) CNS Consortium. METHODS: Methotrexate (8 g/m(2)) was initially administered every 2 weeks. The primary end point was radiographic CR or PR, as defined by standard radiographic criteria, and secondary end points were survival and drug-related toxicity. RESULTS: Twenty five patients were enrolled with a mean age of 60 years and median Karnofsky Performance Score of 80. Three of 14 patients who underwent lumbar puncture had malignant cells on CSF cytopathology, and five of 25 patients had ocular involvement. Two patients could not be evaluated for the primary end point because of the absence of measurable disease in one and death before radiologic imaging in another. All patients have completed the treatment program or progressed. Among 23 patients, there were 12 CR (52%), five PR (22%), one (4%) with stable disease, and five progressions (22%) while on therapy. Seven patients died of tumor progression, and two died of other causes. Median progression-free survival was 12.8 months. Median overall survival for the entire group had not been reached at 22.8+ months. The toxicity of this regimen was modest, with no grade 3 or 4 toxicity in 13 of 25 patients, grade 3 toxicity in eight of 25 patients, and grade 4 toxicity in four of 25 patients after 287 cycles of chemotherapy. CONCLUSION: These results indicate that high-dose methotrexate is associated with modest toxicity and a radiographic response proportion (74%) comparable to more toxic regimens. PMID- 12637470 TI - Randomized phase I/II study of troxacitabine combined with cytarabine, idarubicin, or topotecan in patients with refractory myeloid leukemias. AB - PURPOSE: Troxacitabine has significant single-agent activity. This study was conducted to define the dose-limiting toxicities (DLTs) of its combination with cytarabine (ara-C), idarubicin, or topotecan. PATIENTS AND METHODS: Patients with refractory acute myeloid leukemia (AML), advanced myelodysplastic syndromes (MDS), or chronic myelogenous leukemia in blastic phase (CML-BP) were initially randomly assigned to receive troxacitabine 5.0 mg/m(2) by intravenous (IV) infusion over 30 minutes on days 1 to 5 with ara-C 1.0 g/m(2)/d IV [DOSAGE ERROR CORRECTED] over 2 hours on days 1 to 5, idarubicin 12 mg/m(2) by 5 minute IV infusion on days 1 to 3, or topotecan 1.0 mg/m(2) as an continuous IV infusion on days 1 to 5. Doses were then adjusted to define DLT for each combination. RESULTS: Eighty-seven patients (68 AML, eight MDS, 11 CML-BP) were treated. DLTs were hepatic transaminitis, hyperbilirubinemia, and hand foot syndrome (HFS) on the troxacitabine plus ara-C combination. The recommended phase II doses were 6 mg/m(2) once a day for 5 days and 1.0g/m(2) once a day for 5 days, respectively. DLTs were diarrhea, rash, and mucositis on the troxacitabine plus topotecan combination. The recommended phase II doses were 4 mg/m(2) once a day for 5 days and 0.75 mg/m(2) once a day for 5 days, respectively. DLTs were HFS, rash, and mucositis on the troxacitabine plus idarubicin combination. The recommended phase II doses were 4 mg/m(2) once a day for 5 days and 9 mg/m(2) once a day for 3 days, respectively. Among 74 evaluable patients with AML or MDS, 10 (13%) achieved complete remission and four (5%) had hematologic improvement. Two of 11 (18%) evaluable patients with CML-BP returned to chronic phase. CONCLUSION: Troxacitabine-based combinations had significant antileukemic activity. PMID- 12637471 TI - Clinical outcome of stage I/II melanoma patients after selective sentinel lymph node dissection: long-term follow-up results. AB - PURPOSE: Although sentinel lymph node (SLN) status is part of the new American Joint Committee on Cancer staging system, there is no final proof that the SLN procedure in melanoma patients influences outcome of disease. This study investigated the accuracy of the SLN procedure and clinical outcome in melanoma patients after at least 60 months of follow-up. PATIENTS AND METHODS: Between 1993 and 1996, 209 patients with stage I/II cutaneous melanoma underwent selective SLN dissection by the triple technique. If the SLN contained metastatic disease, a completion lymphadenectomy was performed. Survival analyses were performed using the Kaplan-Meier approach. Factors associated with survival were analyzed using the Cox proportional hazards regression model. RESULTS: The success rate was 99.5%. Median follow-up was 72 months. Forty patients (19%) had a positive SLN. The false-negative rate was 9%. Five-year overall survival was 87% for the entire group and 92% and 67% for SLN-negative and SLN-positive patients (P <.0001), respectively. All patients with a positive SLN and a Breslow thickness < or = 1.00 mm survived, and SLN-positive patients with a Breslow thickness less than 2.00 mm tend to have a better prognosis compared with SLN negative patients with a Breslow thickness greater than 2.00 mm. SLN status (P =.002), Breslow thickness (P =.002), and lymphatic invasion (P =.0009) were all found to be independent prognostic factors for overall survival. CONCLUSION: With a success rate of 99.5% and a false-negative rate of 9% after long-term follow up, the triple-technique SLN procedure is a reliable and accurate method. Survival data seem promising, although a therapeutic effect is still questionable. As shown in this study, not all SLN-positive patients have a poor prognosis. PMID- 12637472 TI - Regulatory approvals of pediatric oncology drugs: previous experience and new initiatives. AB - PURPOSE: To review the Food and Drug Administration (FDA) experience with approvals of new drugs for pediatric oncology and to discuss new regulatory initiatives directed at pediatric oncology. METHODS: A retrospective review of FDA archival documents and the published literature. RESULTS: More than 100 drugs have been approved by the Division of Oncology Drug Products of the FDA for the treatment of malignancies. Only 15 have pediatric use information in their labeling, which is less than 50% of the drugs commonly used in the treatment of pediatric malignancies. In the past 20 years, there have been six submissions to the FDA for pediatric oncology indications. To illustrate principles of the approval process, each submission is discussed. CONCLUSION: Potential reasons for a lack of New Drug Application submissions for pediatric oncology include the small pediatric oncology market compared with the adult oncology market and perceived barriers to performing studies in children. Reasons for failure to approve pediatric indications include small numbers of patients, lack of appropriate controls, and failure to demonstrate patient benefit. Approval criteria include the use of controlled trials, prospective data collection, and disease-appropriate end points. Regulatory initiatives to promote pediatric therapeutic development and product labeling are discussed. PMID- 12637473 TI - Risk of secondary leukemia after a solid tumor in childhood according to the dose of epipodophyllotoxins and anthracyclines: a case-control study by the Societe Francaise d'Oncologie Pediatrique. AB - PURPOSE: To estimate the risk of secondary leukemia as a function of the dose of epipodophyllotoxins and anthracyclines. METHODS: We conducted a case-control study of the risk of secondary leukemia or myelodysplasia after a solid tumor in childhood within the Societe Francaise d'Oncologie Pediatrique, including 61 patients with leukemia matched with 196 controls. The characteristics of the first cancer, the patient's family history of cancer, and the treatment (type, cumulative dose of chemotherapy, schedule of etoposide administration, and radiation dose delivered to active bone marrow) were compared in the two groups. RESULTS: Only two factors were found to increase the risk of leukemia in multivariate analysis, namely, the type of the first tumor, with an excess risk in patients with Hodgkin's disease (relative risk 6.4; 95% confidence interval [CI], 1.6 to 24) or osteosarcoma (relative risk 5; 95% CI, 1.3 to 19), and exposure to epipodophyllotoxins and anthracyclines. The risk of leukemia increased regularly with the cumulative dose of etoposide. In summary, patients who received between 1.2 and 6 g/m(2) of epipodophyllotoxins or more than 170 mg/m(2) of anthracyclines had a seven-fold higher risk (95% CI, 2.6 to 19) compared with patients who received lower doses or none of these drugs. The risk of leukemia in patients who received more than 6 g/m(2) of epipodophyllotoxins was multiplied by 197 (95% CI, 19 to 2,058). The risk of leukemia was not increased by exposure to alkylating agents or radiotherapy. CONCLUSION: Both epipodophyllotoxins and anthracyclines increase the risk of secondary leukemia. The current challenge is to minimize the mutagenic effects of these drugs by diminishing cumulative doses without losing the therapeutic benefits. PMID- 12637474 TI - Impact of metaiodobenzylguanidine scintigraphy on assessing response of high-risk neuroblastoma to dose-intensive induction chemotherapy. AB - PURPOSE: The International Neuroblastoma Response Criteria (INRC) recommend, but do not make mandatory, metaiodobenzylguanidine (MIBG) scans. We present the first report on the effect of MIBG scans on the classification of response to dose intensive induction therapy. PATIENTS AND METHODS: After dose-intensive induction and before consolidative therapy, 162 Memorial Sloan-Kettering Cancer Center (MSKCC) patients with high-risk neuroblastoma (NB) had MIBG scans (99 with (131)I, 63 with (123)I), computed tomography, (99m)Tc-bone scan, bone marrow (BM) tests, and urine catecholamine measurements. Induction included high-dose cyclophosphamide (140 mg/kg) plus other agents and high-dose cisplatin (200 mg/m(2))/etoposide (600 mg/m(2)). RESULTS: In 90 patients treated with dose intensive therapy from diagnosis at MSKCC, the use of MIBG scintigraphy increased the incomplete response numbers from 14 (15.5%) to 20 (22%), giving a complete remission/very good partial remission (CR/VGPR) rate of 78%. In 72 patients treated before referral to MSKCC for intensified therapy, MIBG findings changed the response classification of one patient; the CR/VGPR rate was 43%. MIBG scans showed no BM disease in 15 of 38 patients with histologically evident NB in BM but did show uptake consistent with BM involvement in five patients who had no NB observed in BM tests. CONCLUSION: With the less effective therapy consequent to the intensification of induction only after initial exposure to standard-dose chemotherapy, MIBG scintigraphy merely confirms the findings of other staging modalities for detection of relatively widespread residual NB. However, when dose intensive therapy is initiated at diagnosis, the reliable achievement of major disease responses makes extensive BM testing and MIBG scintigraphy prerequisites for accurate determination of disease status. PMID- 12637476 TI - Early detection of response to radiation therapy in patients with brain malignancies using conventional and high b-value diffusion-weighted magnetic resonance imaging. AB - PURPOSE: To study the feasibility of using diffusion-weighted magnetic resonance imaging (DWMRI), which is sensitive to the diffusion of water molecules in tissues, for detection of early tumor response to radiation therapy; and to evaluate the additional information obtained from high DWMRI, which is more sensitive to low-mobility water molecules (such as intracellular or bound water), in increasing the sensitivity to response. PATIENTS AND METHODS: Standard MRI and DWMRI were acquired before and at regular intervals after initiating radiation therapy for 10 malignant brain lesions in eight patients. RESULTS: One week posttherapy, three of six responding lesions showed an increase in the conventional DWMRI parameters. Another three responding lesions showed no change. Four nonresponding lesions showed a decrease or no change. The early change in the diffusion parameters was enhanced by using high DWMRI. When high DWMRI was used, all responding lesions showed increase in the diffusion parameter and all nonresponding lesions showed no change or decrease. Response was determined by standard MRI 7 weeks posttherapy. The changes in the diffusion parameters measured 1 week after initiating treatment were correlated with later tumor response or no response (P <.006). This correlation was increased to P <.0006 when high DWMRI was used. CONCLUSION: The significant correlation between changes in diffusion parameters 1 week after initiating treatment and later tumor response or no response suggests the feasibility of using DWMRI for early, noninvasive prediction of tumor response. The ability to predict response may enable early termination of treatment in nonresponding patients, prevent additional toxicity, and allow for early changes in treatment. PMID- 12637475 TI - Quantitation of GD2 synthase mRNA by real-time reverse transcriptase polymerase chain reaction: clinical utility in evaluating adjuvant therapy in neuroblastoma. AB - PURPOSE: Minimal residual disease (MRD) is one of the final hurdles to cancer cure. Because therapy (myeloablation, immunotherapy, or differentiation) for MRD is applied at the time of clinical remission, objective surrogate markers are needed to gauge treatment efficacy. PATIENTS AND METHODS: Using quantitative reverse transcriptase polymerase chain reaction (RT-PCR) of GD2 synthase (beta1,4 N-acetylgalactosaminyltransferase, EC 2.4.1.92) mRNA, we evaluated MRD response to anti-GD2 monoclonal antibody 3F8 adjuvant therapy, namely, one cycle of radioimmunotherapy using iodine-131 ((131)I)-3F8 plus one cycle of unlabeled 3F8 in 45 stage 4 neuroblastoma patients (newly diagnosed or without prior relapse) on the N7 protocol at Memorial Sloan-Kettering Cancer Center. The prognostic effect of MRD in their bone marrows before and after this phase of adjuvant therapy on progression-free survival (PFS) and overall survival (OS) was also analyzed. RESULTS: Before 3F8 treatment, 24 of 45 patients were in complete remission (CR), 12 were in very good partial remission (VGPR), and nine were in partial remission (PR), according to criteria from International Neuroblastoma Staging System plus (131)I-3F8 scan; 71% had detectable tumor cells in marrow by real-time RT-PCR. Of the 32 positive patients, 20 became negative after therapy, with a 63% efficacy. When patients were stratified by CR/VGPR versus PR, GD2 synthase positivity was prognostic when detected before 3F8-targeted therapy (PFS, P =.045 and OS, P =.010). Persistent marker positivity was also predictive of PFS (P =.035) and OS (P =.027). Patients who succumbed to the disease had transcript levels four times higher than those who remain alive. CONCLUSION: GD2 synthase mRNA is a useful surrogate marker for evaluating adjuvant treatment efficacy in neuroblastoma with prognostic potential. PMID- 12637477 TI - Radiotherapy for stages IIA/B testicular seminoma: final report of a prospective multicenter clinical trial. AB - PURPOSE: A prospective multicenter trial was initiated to evaluate the role of modern radiotherapy with reduced treatment portals for stage IIA and IIB testicular seminoma. PATIENTS AND METHODS: Patients with stages IIA/B disease (Royal Marsden classification) were assessable for the trial. Staging comprised computed tomography of the chest, abdomen, and pelvis as well as analysis of tumor markers alpha-fetoprotein and beta human chorionic gonadotropin. Linac based radiotherapy was delivered to para-aortic and high ipsilateral iliac lymph nodes. The total doses were 30 Gy for stage IIA and 36 Gy for stage IIB disease. RESULTS: Between April 1991 and March 1994, 94 patients were enrolled for the trial by 30 participating centers throughout Germany. Seven patients were lost to follow-up. Median time to follow-up of 87 assessable patients was 70 months. There were 66 stage IIA and 21 stage IIB patients. One mediastinal and one field edge relapse were observed in the stage IIA group. In the stage IIB group, there was one mediastinal and one mediastinal/pulmonary relapse. All patients were treated with a salvage regimen of platinum-based chemotherapy. Actuarial relapse free survival at 6 years was 95.3% (95% confidence interval [CI], 88.9% to 100%) and 88.9% (95% CI, 74.4% to 100%) for stage IIA and IIB groups, respectively. Maximum acute side effects were 8% grade 3 nausea for stage IIA and 10% grade 3 nausea and diarrhea for stage IIB groups. No late toxicity was observed. CONCLUSION: Radiotherapy for stages IIA/B seminoma with reduced portals yields excellent tumor control at a low rate of acute toxicity and no late toxicity, which supports the role of radiotherapy as the first treatment choice for these patients. PMID- 12637478 TI - Quality of life in good prognosis patients with metastatic germ cell cancer: a prospective study of the European Organization for Research and Treatment of Cancer Genitourinary Group/Medical Research Council Testicular Cancer Study Group (30941/TE20). AB - PURPOSE: To describe global quality of life (GLQL) in patients with metastatic testicular cancer (TC) treated with four different schedules of bleomycin, etoposide, and cisplatin (BEP) chemotherapy (four v three cycles given over 5 v 3 days). PATIENTS AND METHODS: Quality-of-life data were prospectively collected in 666 patients with metastatic TC entered into the European Organization for Research and Treatment of Cancer (EORTC) Trial 30941/United Kingdom Medical Research Council Trial TE20, using the EORTC Quality-of-Life Questionnaire C30 and a TC module. Data were analyzed by a mixed effects model and by evaluation of clinically relevant changes at 2 years. RESULTS: The pattern of GLQL changes was similar in the four groups. Two years after chemotherapy, 36% of patients displayed improved GLQL as compared with baseline, whereas GLQL had deteriorated in 13%. At 3 months, patients receiving the 3-day regimen experienced increased gastrointestinal (GI) toxicity more than those receiving the 5-day regimen, with the difference reaching the level of clinical relevance (>or = 10-point change) if four cycles were given. The 3-day schedule increased the 2-year risk of tinnitus, with clinical relevance demonstrated after four cycles. Long-term peripheral neuropathy and Raynaud-like phenomena were not associated with the number of cycles or days per cycle. At 2 years, Raynaud-like phenomena, tinnitus, or reduced hearing were reported by 21% to 26% of the patients. CONCLUSION: Because of the excess of acute GI toxicity and the increased risk of tinnitus after the 3-day regimen, we recommend the 5-day regimen if four cycles of BEP are planned. If only three cycles are to be given, then the 3-day regimen is acceptable, even given the increased risk of nausea/vomiting at 3 months. PMID- 12637479 TI - Preoperative concurrent chemoradiotherapy in locally advanced rectal cancer with high-dose radiation and oxaliplatin-containing regimen: the Lyon R0-04 phase II trial. AB - PURPOSE: The combination of radiation, fluorouracil, and oxaliplatin in locally advanced rectal cancer has been shown to be feasible in a phase I trial. The purpose of this phase II trial was to assess tolerance and efficacy of this regimen in a preoperative setting. PATIENTS AND METHODS: Between May 2000 and October 2001, 40 operable patients were entered onto the study. Radiotherapy was delivered with a three-field technique to a dose of 50 Gy over 5 weeks with a concomitant boost approach. Two cycles of chemotherapy were given synchronously on weeks 1 and 5, with oxaliplatin 130 mg/m(2) on day 1 followed by 5-day continuous infusion of fluorouracil 350 mg/m(2) and L-folinic acid 100 mg/m(2). Surgery was planned 5 weeks later. RESULTS: All patients completed treatment without modification except one who experienced grade 3/4 toxicity. Grade 3 toxicity was seen in seven patients. Surgery was performed in all patients after a mean interval time of 5 weeks. An objective clinical response was seen in 30 patients (75%). Sphincter-saving surgery was possible in 26 patients. No postoperative deaths occurred. In four patients (10%), a reoperation was necessary (anastomotic fistula, n = 2; pelvic abscess, n = 2). In six cases the operative specimen was sterilized (15%), and in 12 cases (30%), only few residual cells were detected. CONCLUSION: Such a combined preoperative chemoradiotherapy and oxaliplatin-containing regimen is well tolerated with no increase in surgical toxicity. The good response rate observed warrants its use in further clinical trials. PMID- 12637480 TI - Phase I and pharmacokinetic study of intravenous irinotecan plus oral ciclosporin in patients with fuorouracil-refractory metastatic colon cancer. AB - PURPOSE: To assess the safety and toxicity profile of escalating doses of intravenous irinotecan, in combination with a fixed dose of oral ciclosporin (Cs) and to determine the pharmacokinetic profile of irinotecan and its metabolites. PATIENTS AND METHODS: Patients with fluorouracil-refractory metastatic colorectal cancer received escalating doses of intravenous irinotecan from 40 to 125 mg/m(2) every 2 weeks in combination with a fixed dose of oral Cs (5 mg/kg bid for 3 days). Pharmacokinetic analysis of plasma irinotecan and its metabolites SN38 and SN38G was performed during paired cycles with and without Cs. RESULTS: Thirty seven patients were treated. Dose-limiting toxicity of grade 4 neutropenia was seen at an irinotecan dose of 125 mg/m(2). There was no grade 4 diarrhea, and only one patient experienced grade 3 diarrhea. Toxicities caused by Cs were generally mild. Pharmacokinetic studies demonstrated that irinotecan clearance was reduced from 13.4 to 5.8 L/h/m(2) and area under the curve (AUC)(0-tn) was increased 2.2-fold by the coadministration of Cs. Similar significant increases in AUC(0-24h) were seen for both SN38 and SN38G (2.2-fold and 2.3-fold, respectively) in the presence of Cs. Antitumor activity was seen at every irinotecan dose level. CONCLUSION: The maximum tolerated irinotecan dose and recommended dose for phase II studies is 100 mg/m(2) every 2 weeks. Dose-limiting diarrhea was not seen during this study, supporting the hypothesis that pharmacokinetic modulation of irinotecan by Cs may improve its therapeutic index. Further studies using this combination are warranted. PMID- 12637481 TI - Identifying potential indicators of the quality of end-of-life cancer care from administrative data. AB - PURPOSE: To explore potential indicators of the quality of end-of-life services for cancer patients that could be monitored using existing administrative data. METHODS: Quality indicators were identified and assessed by literature review for proposed indicators, focus groups with cancer patients and family members to assess candidate indicators and generate new ideas, and an expert panel ranking the meaningfulness and importance of each potential indicator using a modified Delphi approach. RESULTS: There were three major concepts of poor quality of end of-life cancer care that could be examined using currently-available administrative data (such as Medicare claims): institution of new anticancer therapies or continuation of ongoing treatments very near death; a high number of emergency room visits, inpatient hospital admissions, or intensive care unit days near the end of life; and a high proportion of patients never enrolled in hospice, only admitted in the last few days of life, or dying in an acute-care setting. Concepts such as access to psychosocial and other multidisciplinary services and pain and symptom control are important and may eventually be feasible, but they cannot currently be applied in most data systems. Indicators based on limiting the use of treatments with low probability of benefit or indicators based on economic efficiency were not acceptable to patients, family members, or physicians. CONCLUSION: Several promising claims-based quality indicators were identified that, if found to be valid and reliable within data systems, could be useful in identifying health-care systems in need of improving end-of-life services. PMID- 12637482 TI - Economic evaluation of breast cancer treatment: considering the value of patient choice. AB - PURPOSE: To use 5 years of primary data to compare the incremental cost effectiveness of breast conservation and radiation versus mastectomy with the restriction of choice to a single therapy versus providing a choice of either therapy. PATIENTS AND METHODS: We evaluated a random retrospective cohort of 2,517 Medicare beneficiaries treated for newly diagnosed stage I or II breast cancer from 1992 through 1994. The outcome measures were quality-adjusted life years (QALYs) and 5-year medical costs. Risk and propensity score adjustments were used in the analysis. RESULTS: A breast conservation and radiation regimen has significantly higher costs than mastectomy in the first year after surgery; the adjusted 5-year costs are $14,054 (95% confidence interval, $9,791 to $18,312) greater than those of mastectomy. The adjusted incremental cost effectiveness ratio comparing breast conservation and radiation to mastectomy was $219,594 per QALY for the comparison of the two strategies. If the possibility of patient choice from maintaining the availability of multiple treatments versus restricting choice to mastectomy alone provides a quality-of-life gain of 0.031 QALYs, then the cost-effectiveness ratio of this choice option is $80,440 per QALY. CONCLUSION: The current system of providing a choice between mastectomy and breast conservation surgery is economically attractive when the economic analysis includes the benefit of patient choice of treatment. PMID- 12637484 TI - Maternal preoccupation and parenting as predictors of emotional and behavioral problems in children of women with breast cancer. AB - PURPOSE: To test the hypothesis that differences between sicker and not-so-sick women in their preoccupation with their illness and parenting behavior can explain why some investigators find that children of breast cancer patients fare better than controls and other investigators find the reverse. PATIENTS AND METHODS: Forty-two women with metastasized breast cancer (sicker mothers) and 45 women with a first occurrence of nonmetastasized breast cancer (not-so-sick mothers) rated the degree of their preoccupation with the disease, their parenting behavior, mood, and social supports and the emotional and behavioral symptoms in one of their children. Their 12- to 18-year-old children rated their mothers' parenting behavior, their own emotional and behavioral symptoms, and their self-esteem. RESULTS: Sicker mothers reported relatively less preoccupation. They, and their children, reported less poor parenting and fewer externalizing symptoms in the children. Regression analyses revealed further differences between the groups. CONCLUSION: Less preoccupation with their illness and less poor parenting behavior by sicker mothers may explain why their children seem to fare better then those of not-so-sick mothers. Formulations concerning families of breast cancer patients should include consideration of the effect of the mothers' perception of the severity of their illness. PMID- 12637483 TI - General quality of life 2 years following treatment for prostate cancer: what influences outcomes? Results from the prostate cancer outcomes study. AB - PURPOSE: The goal of this study was to determine the relationship between primary treatment, urinary dysfunction, sexual dysfunction, and general health-related quality of life (HRQOL) in prostate cancer. METHODS: A sample of men with newly diagnosed prostate cancer between 1994 and 1995 was randomly selected from six population-based Surveillance, Epidemiology, and End Results registries. A baseline survey was completed by 2,306 men within 6 to 12 months of diagnosis, and these men also completed a follow-up HRQOL survey 2 years after diagnosis. Logistic regression models were used to determine whether primary treatment, urinary dysfunction, and sexual dysfunction were independently associated with general HRQOL outcomes approximately 2 years after diagnosis as measured by the Medical Outcomes Study 36-item Short Form Health Survey. The magnitude of this effect was estimated using least square means models. RESULTS: After adjustment for potential confounders, primary treatment was not associated with 2-year general HRQOL outcomes in men with prostate cancer. Urinary function and bother were independently associated with worse general HRQOL in all domains. Sexual function and bother were also independently associated with worse general HRQOL, although the relationship was not as strong as in the urinary domains. CONCLUSION: Primary treatment is not associated with 2-year general HRQOL outcomes in prostate cancer. Although both sexual and urinary function and bother are associated with quality of life, men who are more bothered by their urination or impotence are more likely to report worse quality of life. This implies that future research should be directed toward finding ways to improve treatment related outcomes or help patients better cope with their posttreatment urinary or sexual dysfunction. PMID- 12637485 TI - Influenza vaccination in elderly patients with advanced colorectal cancer. AB - PURPOSE: To examine influenza vaccination use in patients undergoing chemotherapy for advanced cancer. METHODS: All Medicare patients treated for stage IV colorectal cancer between 1993 and 1998 while living in one of the regions monitored by the Survival, Epidemiology, and End Results Program who were alive in the fall months and who survived at least 4 months with their cancer were considered eligible to have received vaccination. Their medical bills were analyzed to determine receipt of influenza vaccination and subsequent outcomes. RESULTS: Eligibility criteria were met by 1,225 patients who were undergoing chemotherapy during 1,577 person-years of observation. Overall, 39.7% of patients received influenza vaccination, increasing from 26% in 1993 to 43% in 1998. When vaccination was administered, it was provided by primary care physicians 68% of the time. Vaccinated patients were more likely to be white, of higher socioeconomic status, and to have more comorbidity. Fewer diagnoses of influenza and pneumonia infections were made in vaccinated patients while undergoing treatment. Those patients who were immunized also had fewer chemotherapy interruptions and were more likely to survive through to the beginning of the next fall (hazard ratio, 0.88; 95% confidence interval, 0.77 to 0.99). There was a trend toward decreased resource use among immunized patients. CONCLUSION: This study observed outcomes associated with influenza vaccination that are similar to those reported for patients without cancer. However, rates of immunization are relatively low, and disparities exist for vulnerable populations. As part of delivering high-quality care, oncologists should promote influenza vaccination for their patients who are undergoing treatment for advanced cancer. PMID- 12637486 TI - Phase I clinical and pharmacology study of clofarabine in patients with solid and hematologic cancers. AB - PURPOSE: To define the maximum-tolerated doses (MTDs) and dose-limiting toxicities (DLTs) of clofarabine, given as a 1-hour infusion daily for 5 days, in patients with solid tumors and with acute leukemia. PATIENTS AND METHODS: The initial part of the study defined the MTD and DLT in solid tumors. The second part of the study defined the MTD and DLT in acute leukemia. RESULTS: The starting dose of clofarabine (15 mg/m(2)) was myelosuppressive, requiring several dose de-escalations to 2 mg/m(2), the dose suggested for phase II studies in solid tumors. Dose escalation in acute leukemia started at 7.5 mg/m(2), with several escalations to 55 mg/m(2). The DLT was reversible hepatotoxicity at 55 mg/m(2). The recommended dose for acute leukemia phase II studies was 40 mg/m(2). Among 32 treated patients with acute leukemia, two achieved a complete response and three had a marrow complete response without platelet recovery (hematologic improvement), for an overall response rate of 16%. At 40 mg/m(2), the median plasma clofarabine level was 1.5 micro mol/L (range, 0.42 to 3.2 micro mol/L; n = 7). Cellular and plasma pharmacokinetic studies suggested dose proportionality but showed a wide variation in intracellular concentrations of clofarabine triphosphate. CONCLUSION: This phase I study defined the following two MTDs for clofarabine given as a 1-hour infusion daily for 5 days: 2 mg/m(2) for solid tumors, the DLT being myelosuppression; and 40 mg/m(2) for acute leukemia, the DLT being hepatotoxicity. Encouraging activity was observed in acute leukemia. PMID- 12637487 TI - Role of DNA mismatch repair defects in the pathogenesis of human cancer. AB - The DNA mismatch repair (MMR) system is necessary for the maintenance of genomic stability. In a broad sense, all main functions of the MMR system, including the correction of biosynthetic errors, DNA damage surveillance, and prevention of recombination between nonidentical sequences serve this important purpose. Failure to accomplish these functions may lead to cancer. It is therefore not surprising that inherited defects in the MMR system underlie one of the most prevalent cancer syndromes in humans, hereditary nonpolyposis colon cancer (HNPCC). In addition, acquired defects of the same system may account for 15% to 25%, or even a higher percentage, of sporadic cancers of different organs of the "HNPCC spectrum," including the colon and rectum, uterine endometrium, stomach, and ovaries. Recent studies indicate that the MMR genes may be involved in the pathogenesis of even a broader spectrum of tumors in one way or another. An updated review of the different features of the human MMR system will be provided, with the emphasis on their implications in cancer development. PMID- 12637488 TI - Granulosa cell tumor of the ovary. AB - Adult granulosa cell tumor (GCT) of the ovary is oftentimes a hormonally active, stromal cell neoplasm that is distinguished by its ability to secrete sex steroids such as estrogen. Patients may present with vaginal bleeding caused by endometrial hyperplasia or uterine cancer as a result of prolonged exposure to tumor-derived estrogen. In addition, GCT is a vascular tumor that may occasionally rupture and result in abdominal pain, hemoperitoneum, and hypotension, mimicking an ectopic pregnancy in younger patients. GCT is usually associated with a mass on pelvic examination that is subsequently confirmed on ultrasonography. Surgery is required for definitive tissue diagnosis, staging, and tumor debulking. In older women, a total abdominal hysterectomy and bilateral salpingooophorectomy are typically performed. In women of childbearing age, a more conservative unilateral salpingo-oophorectomy may be performed, assuming that careful staging reveals that the disease has not extended outside of the involved ovary and that a concomitant uterine cancer has been excluded. Survival of patients with GCT is generally excellent because most patients present with early-stage disease, although certain high-risk patient groups may be identified. Stage is the most important prognostic factor, with a higher risk of relapse being associated with stages II through IV disease. In addition, patients with stage I disease associated with features such as large tumor size, high mitotic index, or tumor rupture may also be at higher risk in some series. The value of postoperative adjuvant therapy for high-risk patients has not been investigated by prospective randomized trials, which are difficult to perform because of the rarity of this tumor. Nonetheless, the use of adjuvant chemotherapy or radiation has sometimes been associated with prolonged disease-free survival in patients with high-risk features. Because of the propensity of GCT to recur years after initial diagnosis, prolonged surveillance with serial physical examination and serum tumor markers such as estradiol and inhibin is reasonable. PMID- 12637489 TI - Adjuvant chemotherapy for node-positive breast cancer patients: which is the reference today? PMID- 12637492 TI - Randomized comparison of low-dose involved-field radiotherapy and no radiotherapy for children with Hodgkin's disease who achieve a complete response to chemotherapy, by Nachman et al. PMID- 12637493 TI - CD40-dependent activation of phosphatidylinositol 3-kinase/Akt pathway mediates endothelial cell survival and in vitro angiogenesis. AB - CD40 has been involved in tumor and inflammatory neoangiogenesis. In this study we determined that stimulation of endothelial CD40 with sCD154 induced resistance to apoptosis and in vitro vessel-like formation by human microvascular endothelial cells (HMEC). These effects were determined to be mediated by CD40 dependent signaling because they were inhibited by a soluble CD40-muIg fusion protein. Moreover, apoptosis of HMEC was associated with an impairment of Akt phosphorylation, which was restored by stimulation with sCD154. The anti apoptotic effect as well as in vitro vessel-like formation and Akt phosphorylation were inhibited by treatment of HMEC with two unrelated pharmacological inhibitors of phosphatidylinositol 3-kinase (PI3K), wortmannin and LY294002. CD40 stimulation induced a rapid increase in Akt enzymatic activity that was not prevented by cycloheximide, an inhibitor of protein synthesis. The enhanced Akt activity induced by stimulation of endothelial CD40 was temporarily correlated with the association of CD40 with TRAF6, c-Cbl, and the p85 subunit of PI3K. Expression of negative-dominant Akt inhibited the activation of endogenous Akt through CD40 stimulation, despite the observation that association of CD40 with TRAF6, c-Cbl, and PI3K was intact. The defective activation of Akt abrogated not only the anti-apoptotic effect of CD40 stimulation but also the proliferative response, the enhanced motility, and the in vitro formation of vessel-like tubular structures by CD40-stimulated HMEC. In conclusion, these results suggest that endothelial CD40, through activation of the PI3K/Akt signaling pathway, regulates cell survival, proliferation, migration, and vessel-like structure formation, all steps considered critical for angiogenesis. PMID- 12637495 TI - Analysis of the interaction of small heat shock proteins with unfolding proteins. AB - The ubiquitous small heat shock proteins (sHsps) are efficient molecular chaperones that interact with nonnative proteins, prevent their aggregation, and support subsequent refolding. No obvious substrate specificity has been detected so far. A striking feature of sHsps is that they form large complexes with nonnative proteins. Here, we used several well established model chaperone substrates, including citrate synthase, alpha-glucosidase, rhodanese, and insulin, and analyzed their interaction with murine Hsp25 and yeast Hsp26 upon thermal unfolding. The two sHsps differ in their modes of activation. In contrast to Hsp25, Hsp26 undergoes a temperature-dependent dissociation that is required for efficient substrate binding. Our analysis shows that Hsp25 and Hsp26 reacted in a similar manner with the nonnative proteins. For all substrates investigated, complexes of defined size and shape were formed. Interestingly, several different nonnative proteins could be incorporated into defined sHsp-substrate complexes. The first substrate protein bound seems to determine the complex morphology. Thus, despite the differences in quaternary structure and mode of activation, the formation of large uniform sHsp-substrate complexes seems to be a general feature of sHsps, and this unique chaperone mechanism is conserved from yeast to mammals. PMID- 12637494 TI - Glucocorticoids inhibit apoptosis during fibrosarcoma development by transcriptionally activating Bcl-xL. AB - Glucocorticoids influence many physiological processes, and in particular apoptosis, often with opposite effects depending on the cell type examined. We found that during fibrosarcoma development there is a strong increase in apoptosis at the tumor stage, which is repressed by dexamethasone to levels observed in normal fibroblasts. The anti-apoptotic Bcl-2 family protein Bcl-x(L) is induced by dexamethasone at the transcriptional level at all stages of fibrosarcoma development. The ligand-activated glucocorticoid receptor (GR) activates the Bcl-x promoter in transient transfection experiments, and GR binds to specific Bcl-x promoter sequences in vitro and in vivo. Furthermore, a GR antagonist abolishes this effect, indicating that Bcl-x(L) induction is mediated by GR. Importantly, exogenous Bcl-x(L) inhibits apoptosis and caspase-3 activity in fibrosarcoma cells to levels found in dexamethasone-treated fibrosarcoma cells. We conclude that Bcl-x(L) is a key target mediating the anti-apoptotic effects of glucocorticoids during fibrosarcoma development. These observations provide further understanding of the molecular basis of glucocorticoid regulation of cell death during tumorigenesis. PMID- 12637496 TI - Electrogenicity of Na,K- and H,K-ATPase activity and presence of a positively charged amino acid in the fifth transmembrane segment. AB - The transport activity of the Na,K-ATPase (a 3 Na+ for 2 K+ ion exchange) is electrogenic, whereas the closely related gastric and non-gastric H,K-ATPases perform electroneutral cation exchange. We have studied the role of a highly conserved serine residue in the fifth transmembrane segment of the Na,K-ATPase, which is replaced with a lysine in all known H,K-ATPases. Ouabain-sensitive 86Rb uptake and K+-activated currents were measured in Xenopus oocytes expressing the Bufo bladder H,K-ATPase or the Bufo Na,K-ATPase in which these residues, Lys800 and Ser782, respectively, were mutated. Mutants K800A and K800E of the H,K-ATPase showed K+-stimulated and ouabain-sensitive electrogenic transport. In contrast, when the positive charge was conserved (K800R), no K+-induced outward current could be measured, even though rubidium transport activity was present. Conversely, the S782R mutant of the Na,K-ATPase had non-electrogenic transport activity, whereas the S782A mutant was electrogenic. The K800S mutant of the H,K ATPase had a more complex behavior, with electrogenic transport only in the absence of extracellular Na+. Thus, a single positively charged residue in the fifth transmembrane segment of the alpha-subunit can determine the electrogenicity and therefore the stoichiometry of cation transport by these ATPases. PMID- 12637497 TI - Structures of shikimate dehydrogenase AroE and its Paralog YdiB. A common structural framework for different activities. AB - Shikimate dehydrogenase catalyzes the fourth step of the shikimate pathway, the essential route for the biosynthesis of aromatic compounds in plants and microorganisms. Absent in metazoans, this pathway is an attractive target for nontoxic herbicides and drugs. Escherichia coli expresses two shikimate dehydrogenase paralogs, the NADP-specific AroE and a putative enzyme YdiB. Here we characterize YdiB as a dual specificity quinate/shikimate dehydrogenase that utilizes either NAD or NADP as a cofactor. Structures of AroE and YdiB with bound cofactors were determined at 1.5 and 2.5 A resolution, respectively. Both enzymes display a similar architecture with two alpha/beta domains separated by a wide cleft. Comparison of their dinucleotide-binding domains reveals the molecular basis for cofactor specificity. Independent molecules display conformational flexibility suggesting that a switch between open and closed conformations occurs upon substrate binding. Sequence analysis and structural comparison led us to propose the catalytic machinery and a model for 3-dehydroshikimate recognition. Furthermore, we discuss the evolutionary and metabolic implications of the presence of two shikimate dehydrogenases in E. coli and other organisms. PMID- 12637499 TI - Purified, recombinant TagF protein from Bacillus subtilis 168 catalyzes the polymerization of glycerol phosphate onto a membrane acceptor in vitro. AB - We report the first characterization of a recombinant protein involved in the polymerization of wall teichoic acid. Previously, a study of the teichoic acid polymerase activity associated with membranes from Bacillus subtilis 168 strains bearing thermosensitive mutations in tagB, tagD, and tagF implicated TagF as the poly(glycerol phosphate) polymerase (Pooley, H. M., Abellan, F. X., and Karamata, D. (1992) J. Bacteriol. 174, 646-649). In the work reported here, we have demonstrated an unequivocal role for tagF in the thermosensitivity of one such mutant (tagF1) by conditional complementation at the restrictive temperature with tagF under control of the xylose promoter at the amyE locus. We have overexpressed and purified recombinant B. subtilis TagF protein, and we provide direct biochemical evidence that this enzyme is responsible for polymerization of poly(glycerol phosphate) teichoic acid in B. subtilis 168. Recombinant hexahistidine-tagged TagF protein was purified from Escherichia coli and was used to develop a novel membrane pelleting assay to monitor poly(glycerol phosphate) polymerase activity. Purified TagF was shown to incorporate radioactivity from its substrate CDP-[(14)C]glycerol into a membrane fraction in vitro. This activity showed a saturable dependence on the concentration of CDP-glycerol (K(m) of 340 microm) and the membrane acceptor (half-maximal activity at 650 microg of protein/ml of purified B. subtilis membranes). High pressure liquid chromatography analysis confirmed the polymeric nature of the reaction product, approximately 35 glycerol phosphate units in length. PMID- 12637498 TI - The role of the Ah receptor and p38 in benzo[a]pyrene-7,8-dihydrodiol and benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide-induced apoptosis. AB - Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous contaminants in the environment. Benzo[a]pyrene (B[a]P), a prototypical member of this class of chemicals, affects cellular signal transduction pathways and induces apoptosis. In this study, the proximate carcinogen of B[a]P metabolism, trans-7,8-dihydroxy 7,8-dihydrobenzo[a]pyrene (B[a]P-7,8-dihydrodiol) and the ultimate carcinogen, B[a]P-r-7,t-8-dihydrodiol-t-9,10-epoxide(+/-) (BPDE-2) were found to induce apoptosis in human HepG2 cells. Apoptosis initiated by B[a]P-7,8-dihydrodiol was linked to activation of the Ah receptor and induction of CYP1A1, an event that can lead to the formation of BPDE-2. With both B[a]P-7,8-dihydrodiol and BPDE-2 treatment, changes in anti- and pro-apoptotic events in the Bcl-2 family of proteins correlated with the release of mitochondrial cytochrome c and caspase activation. The onset of apoptosis as monitored by caspase activation was linked to mitogen-activated protein (MAP) kinases. Utilizing mouse hepa1c1c7 cells and the Arnt-deficient BPRc1 cells, activation of MAP kinase p38 by B[a]P-7,8 dihydrodiol was shown to be Ah receptor-dependent, indicating that metabolic activation by CYP1A1 was required. This was in contrast to p38 activation by BPDE 2, an event that was independent of Ah receptor function. Confirmation that MAP kinases play a critical role in BPDE-2-induced apoptosis was shown by inhibiting caspase activation of poly(ADP-ribose)polymerase 1 (PARP-1) by chemical inhibitors of p38 and ERK1/2. Furthermore, mouse embryo p38-/- fibroblasts were shown to be resistant to the actions of BPDE-2-induced apoptosis as determined by annexin V analysis, cytochrome c release, and cleavage of PARP-1. These results confirm that the Ah receptor plays a critical role in B[a]P-7,8-dihydrodiol induced apoptosis while p38 MAP kinase links the actions of an electrophilic metabolite like BPDE-2 to the regulation of programmed cell death. PMID- 12637500 TI - The activity of Mblk-1, a mushroom body-selective transcription factor from the honeybee, is modulated by the ras/MAPK pathway. AB - We previously identified a gene, termed Mblk-1, that encodes a putative transcription factor with two DNA-binding motifs expressed preferentially in the mushroom body of the honeybee brain, and its preferred binding sequence, termed Mblk-1-binding element (MBE) (Takeuchi, H., Kage, E., Sawata, M., Kamikouchi, A., Ohashi, K., Ohara, M., Fujiyuki, T., Kunieda, T., Sekimizu, K., Natori, S., and Kubo, T. (2001) Insect Mol Biol 10, 487-494; Park, J.-M., Kunieda. T., Takeuchi, H., and Kubo, T. (2002) Biochem. Biophys. Res. Commun. 291, 23-28). In the present study, the effect of Mblk-1 on transcription of genes containing MBE in Drosophila Schneider's Line 2 cells was examined using a luciferase assay. Mblk-1 expression transactivated promoters containing MBEs approximately 2-7-fold. Deletion experiments revealed that RHF2, the second DNA-binding domain of Mblk-1, was necessary for the transcriptional activity. Furthermore, mitogen-activated protein kinase (MAPK) phosphorylated Mblk-1 at Ser-444 in vitro, and the Mblk-1 induced transactivation was stimulated by phosphorylation of Ser-444 by the Ras/MAPK pathway in the luciferase assay. These results suggest that Mblk-1 is a transcription factor that might function in the mushroom body neuronal circuits downstream of the Ras/MAPK pathway in the honeybee brain. PMID- 12637501 TI - SufA from Erwinia chrysanthemi. Characterization of a scaffold protein required for iron-sulfur cluster assembly. AB - SufA is a component of the recently discovered suf operon, which has been shown to play an important function in bacteria during iron-sulfur cluster biosynthesis and resistance to oxidative stress. The SufA protein from Erwinia chrysanthemi, a Gram-negative plant pathogen, has been purified to homogeneity and characterized. It is a homodimer with the ability to assemble rather labile [2Fe-2S] and [4Fe 4S] clusters as shown by Mossbauer spectroscopy. These clusters can be transferred to apoproteins such as ferredoxin or biotin synthase during a reaction that is not inhibited by bathophenanthroline, an iron chelator. Cluster assembly in these proteins is much more efficient when iron and sulfur are provided by holoSufA than by free iron sulfate and sodium sulfide. We propose the function of SufA is that of a scaffold protein for [Fe-S] cluster assembly and compare it to IscA, a member of the isc operon also involved in cluster biosynthesis in both prokaryotes and eukaryotes. Mechanistic and physiological implications of these results are also discussed. PMID- 12637503 TI - Allosteric modulation of ligand binding to low density lipoprotein receptor related protein by the receptor-associated protein requires critical lysine residues within its carboxyl-terminal domain. AB - The low density lipoprotein receptor-related protein (LRP) is a large endocytic receptor that recognizes more than 30 different ligands and plays important roles in protease and lipoprotein catabolism. Ligand binding to newly synthesized LRP is modulated by the receptor-associated protein (RAP), an endoplasmic reticulum resident protein that functions as a molecular chaperone and prevents ligands from associating with LRP via an allosteric-type mechanism. RAP is a multidomain protein that contains two independent LRP binding sites, one located at the amino terminal portion of the molecule and the other at the carboxyl-terminal portion of the molecule. The objective of the present investigation was to gain insight into how these two regions of RAP interact with LRP and function to modulate its ligand binding properties. These objectives were accomplished by random mutagenesis of RAP, which identified two critical lysine residues, Lys-256 and Lys-270, within the carboxyl-terminal domain that are necessary for binding of this region of RAP to LRP and to heparin. RAP molecules in which either of these two lysine residues was mutated still bound LRP but with reduced affinity. Furthermore, the mutant RAPs were significantly impaired in their ability to inhibit alpha(2)M* binding to LRP via allosteric mechanisms. In contrast, the mutant RAP molecules were still effective at inhibiting uPA.PAI-1 binding to LRP. These results confirm that both LRP binding sites within RAP cooperate to inhibit ligand binding via an allosteric mechanism. PMID- 12637502 TI - Regulation of epidermal growth factor-induced connexin 43 gap junction communication by big mitogen-activated protein kinase1/ERK5 but not ERK1/2 kinase activation. AB - The gap junction protein, Cx43, plays a pivotal role in coupling cells electrically and metabolically, and the putative phosphorylation sites that modulate its function are reflected as changes in gap junction communication. Growth factor stimulation has been correlated with a decrease in gap junction communication and a parallel activation of ERK1/2; the inhibition of epidermal growth factor (EGF)-induced Cx43 gap junction uncoupling was observed by using the MEK1/2 inhibitor, PD98059. Because 1) BMK1/ERK5, another MAPK family member also activated by growth factors, possesses a phosphorylation motif similar to ERK1/2, and 2) it has been reported that PD98059 can inhibit not only MEK1/2 ERK1/2 but also MEK5-BMK1 activation, we investigated whether BMK1 can regulate EGF-induced Cx43 gap junction uncoupling and phosphorylation, comparing this to the role of ERK1/2 on Cx43 function and phosphorylation induced by EGF. Selective activation or inactivation of ERK1/2 by using a constitutively active form or a dominant negative form of MEK1 did not regulate Cx43 gap junction coupling. In contrast, we found that BMK1, selectively activated by constitutively active MEK5alpha, induced gap junction uncoupling, and the inhibition of BMK1 activation by transfection of dominant negative BMK1 prevented EGF-induced gap junction uncoupling. Activated BMK1 selectively phosphorylates Cx43 on Ser-255 in vitro and in vivo, but not on S279/S282, which are reported as the consensus phosphorylation sites for MAPK. Furthermore, by co-immunoprecipitation, we found that BMK1 directly associates with Cx43 in vivo. These data indicate that BMK1 is more important than ERK1/2 in EGF-mediated Cx43 gap junction uncoupling by association and Cx43 Ser- 255 phosphorylation. PMID- 12637504 TI - Two amino acid residues determine the low substrate affinity of human cationic amino acid transporter-2A. AB - Mammalian cationic amino acid transporters (CAT) differ in their substrate affinity and sensitivity to trans-stimulation. The apparent Km values for cationic amino acids and the sensitivity to trans-stimulation of CAT-1, -2B, and 3 are characteristic of system y+. In contrast, CAT-2A exhibits a 10-fold lower substrate affinity and is largely independent of substrate at the trans-side of the membrane. CAT-2A and -2B demonstrate such divergent transport properties, even though their amino acid sequences differ only in a stretch of 42 amino acids. Here, we identify two amino acid residues within this 42-amino acid domain of the human CAT-2A protein that are responsible for the apparent low affinity of both the extracellular and intracellular substrate-binding sites. These residues are located in the fourth intracellular loop, suggesting that they are not part of the translocation pathway. Rather, they may be responsible for the low affinity conformation of the substrate-binding sites. The sensitivity to trans stimulation is not determined by the same amino acid residues as the substrate affinity and must involve a more complex interaction between individual amino acid residues. In addition to the 42-amino acid domain, the adjacent transmembrane domain X seems to be involved in this function. PMID- 12637506 TI - Apolipoprotein A5, a crucial determinant of plasma triglyceride levels, is highly responsive to peroxisome proliferator-activated receptor alpha activators. AB - The recently discovered APOA5 gene has been shown in humans and mice to be important in determining plasma triglyceride levels, a major cardiovascular disease risk factor. apoAV represents the first described apolipoprotein where overexpression lowers triglyceride levels. Since fibrates represent a commonly used therapy for lowering plasma triglycerides in humans, we investigated their ability to modulate APOA5 gene expression and consequently influence plasma triglyceride levels. Human primary hepatocytes treated with Wy 14,643 or fenofibrate displayed a strong induction of APOA5 mRNA. Deletion and mutagenesis analyses of the proximal APOA5 promoter firmly demonstrate the presence of a functional peroxisome proliferator-activated receptor response element. These findings demonstrate that APOA5 is a highly responsive peroxisome proliferator activated receptor alpha target gene and support its role as a major mediator for how fibrates reduce plasma triglycerides in humans. PMID- 12637505 TI - Sustained activation of JNK/p38 MAPK pathways in response to cisplatin leads to Fas ligand induction and cell death in ovarian carcinoma cells. AB - The efficacy of cisplatin in cancer chemotherapy is limited by the development of resistance. Although the molecular mechanisms involved in chemoresistance are poorly understood, cellular response to cisplatin is known to involve activation of MAPK and other signal transduction pathways. An understanding of early signal transduction events in the response to cisplatin could be valuable for improving the efficacy of cancer therapy. We compared cisplatin-induced activation of three MAPKs, JNK, p38, and ERK, in a cisplatin-sensitive human ovarian carcinoma cell line (2008) and its resistant subclone (2008C13). The JNK and p38 pathways were activated differentially in response to cisplatin, with the cisplatin-sensitive cells showing prolonged activation (8-12 h) and the cisplatin-resistant cells showing only transient activation (1-3 h) of JNK and p38. In the sensitive cells, inhibition of cisplatin-induced JNK and p38 activation blocked cisplatin-induced apoptosis; persistent activation of JNK resulted in hyperphosphorylation of the c Jun transcription factor, which in turn stimulated the transcription of an immediate downstream target, the death inducer Fas ligand (FasL). Sequestration of FasL by incubation with a neutralizing anti-FasL antibody inhibited cisplatin induced apoptosis. In contrast, chemoresistance in 2008C13 cells was associated with failure to up-regulate FasL. Moreover, in these cells, selective stimulation of the JNK/p38 MAPK pathways by adenovirus-mediated delivery of recombinant MKK7 or MKK3 led to sensitization to apoptosis through reactivating FasL expression. Thus, the JNK > c-Jun > FasL > Fas pathway plays an important role in mediating cisplatin-induced apoptosis in ovarian cancer cells, and the duration of JNK activation is critical in determining whether cells survive or undergo apoptosis. PMID- 12637507 TI - p53 inhibitor pifithrin alpha can suppress heat shock and glucocorticoid signaling pathways. AB - Pifithrin alpha (PFTalpha) is a chemical compound isolated for its ability to suppress p53-mediated transactivation. It can protect cells from p53-mediated apoptosis induced by various stimuli and reduce sensitivity of mice to gamma radiation. Identification of molecular targets of PFTalpha is likely to provide new insights into mechanisms of regulation of p53 pathway and is important for predicting potential risks associated with administration of PFTalpha-like p53 inhibitors in vivo. We found that PFTalpha, in addition to p53, can suppress heat shock and glucocorticoid receptor signaling but has no effect on nuclear factor kappaB signaling. PFTalpha reduces activation of heat shock transcription factor (HSF1) and increases cell sensitivity to heat. Moreover, it reduces activation of glucocorticoid receptor and rescues mouse thymocytes in vitro and in vivo from apoptotic death after dexamethasone treatment. PFTalpha affected both signaling pathways in a p53-independent manner. These observations suggest that PFTalpha targets some unknown factor that is common for three major signal transduction pathways. PMID- 12637509 TI - A Novel voltage-dependent chloride current activated by extracellular acidic pH in cultured rat Sertoli cells. AB - Sertoli cells from mammalian testis are key cells involved in development and maintenance of stem cell spermatogonia as well as secretion of a chloride- and potassium-rich fluid into the lumen of seminiferous tubules. Using whole-cell patch clamp experiments, a novel chloride current was identified. It is activated only in the presence of an extracellular acidic pH, with an estimated half maximal activation at pH 5.5. The current is strongly outwardly rectifying, activated with a fast time-dependent onset of activation but a slow time dependent kinetic at depolarization pulses. The pH-activated chloride current was not detected at physiological or basic pH and is not sensitive to intracellular or extracellular Ca2+ variation. Diphenylamine-2-carboxylic acid and 4,4' diisothiocyanatostilbene-2,2'-disulfonic acid blocked the induced currents, and its anionic selectivity sequence was Cl- > Br- > I-> gluconate. We have performed a reverse transcription-PCR analysis to search for voltage-dependent chloride rClC channels in cultured rat Sertoli cells. Among the nine members of the family only rClC-2, rClC-3, rClC-6, and rClC-7 have been identified. The inwardly rectifying rClC-2 chloride current was activated by hyperpolarization but not by pH variation. A different depolarization-activated outwardly rectifying chloride current was activated only by hypotonic challenge and may correspond either to rClC-3 or rClC-6. Immunolocalization experiments demonstrate that rClC-7 resides in the intracellular compartment of Sertoli cells. This study provides the first functional identification of a native acid-activated chloride current. Based on our molecular analysis of rClC proteins, this new chloride current does not correspond to rClC-2, rClC-3, rClC-6, or rClC-7 channels. The potential physiological role of this native current in an epithelial cell from the reproductive system is discussed. PMID- 12637508 TI - The cell adhesion molecule CEACAM1-L is a substrate of caspase-3-mediated cleavage in apoptotic mouse intestinal cells. AB - The CEACAM1 cell adhesion molecule is a member of the carcinoembryonic antigen family. In the mouse, four distinct isoforms are generated by alternative splicing. These encode either two or four immunoglobulin domains linked through a transmembrane domain to a cytoplasmic domain that encompasses either a short 10 amino acid tail or a longer one of 73 amino acids. Inclusion of exon 7, well conserved in evolution, generates the long cytoplasmic domain. A potential caspase recognition site in mouse, rat, and human CEACAM1-L also becomes available within the peptide encoded by exon 7. We used CEACAM1-L-transfected mouse colon carcinoma CT51 cells treated with three different apoptotic agents to study its fate during cell death. We found that CEACAM1-L is cleaved resulting in rapid degradation of most of its 8-kDa cytoplasmic domain. Caspase-mediated cleavage was demonstrated using purified recombinant caspases. The long cytoplasmic domain was cleaved specifically by caspase-3 in vitro but not by caspase-7 or -8. Moreover cleavage of CEACAM1-L in apoptotic cells was blocked by addition of a selective caspase-3 inhibitor to the cultures. Using point and deletion mutants, the conserved DQRD motif in the membrane-proximal cytoplasmic domain was identified as a caspase cleavage site. We also show that once CEACAM1 L is caspase-cleaved it becomes a stronger adhesion molecule than both the shorter and the longer expressing isoforms. PMID- 12637510 TI - Shear flow attenuates serum-induced STAT3 activation in endothelial cells. AB - Vascular endothelial cells (ECs) are constantly exposed to flow-induced shear stress. Shear stress is known to induce signaling cascades, including the extracellular signal-regulated protein kinase (ERK) pathway. STAT3 transcription factor plays a key role in cytokine stimulation. Recent studies indicate that STAT3 is involved in growth factor-induced cell cycle. In the present study, we have examined STAT3 activation of ECs under conditions of shear flow. Bovine aortic ECs cultured with serum at static state show a serum concentration dependent phosphorylation at Tyr-705 of STAT3, whereas there is a constant basal phosphorylation at Ser-727. In ECs subjected to shear flow, a shear dose dependent phosphorylation of Ser-727 and ERK1/2 was observed. In contrast, a concomitantly shear dose-dependent inhibition of phosphorylation at Tyr-705 was exhibited. Shear stress on ECs increased the association of ERK1/2 to STAT3. ECs treated with MEK inhibitor (U0126 or PD98059) consistently and significantly reduced the shear-induced ERK1/2 and Ser-727 phosphorylation, indicating that ERK1/2 is upstream of Ser-727 phosphorylation. Interestingly, shear-induced inhibition in Tyr-705 phosphorylation was abolished in these same inhibitor treated ECs. Similarly, ECs transfected with a dominant positive mutant of MEK1 enhanced the phosphorylation of Ser-727 with the attenuation of the Tyr-705 phosphorylation. In contrast, when ECs were transfected with dominant positive mutant of MEKK1, JNK upstream, no change in the phosphorylation of Ser-727 and Tyr-705 was observed. These results indicate that shear flow induces the phosphorylation of Ser-727 via ERK1/2 pathway, and this Ser-727 phosphorylation inhibits Tyr-705 phosphorylation in STAT3. As a result, shear flow reduced the translocation of STAT3 into nucleus. This study shows for the first time that shear flow may play a significant role by attenuating STAT3 activation and thus may reduce inflammatory responses and/or serum-induced endothelial proliferation. PMID- 12637511 TI - Specific beta1 integrin site selectively regulates Akt/protein kinase B signaling via local activation of protein phosphatase 2A. AB - Integrin transmembrane receptors generate multiple signals, but how they mediate specific signaling is not clear. Here we test the hypothesis that particular sequences along the beta(1) integrin cytoplasmic domain may exist that are intimately related to specific integrin-mediated signaling pathways. Using systematic alanine mutagenesis of amino acids conserved between different beta integrin cytoplasmic domains, we identified the tryptophan residue at position 775 of human beta(1) integrin as specific and necessary for integrin-mediated protein kinase B/Akt survival signaling. Stable expression of a beta(1) integrin mutated at this amino acid in GD25 beta(1)-null cells resulted in reduction of Akt phosphorylation at both Ser(473) and Thr(308) activation sites. As a consequence, the cells were substantially more sensitive to serum starvation induced apoptosis when compared with cells expressing wild type beta(1) integrin. This inactivation of Akt resulted from increased dephosphorylation by a localized active population of protein phosphatase 2A. Both Akt and protein phosphatase 2A were present in beta(1) integrin-organized cytoplasmic complexes, but the activity of this phosphatase was 2.5 times higher in the complexes organized by the mutant integrin. The mutation of Trp(775) specifically affected Akt signaling, without effects on other integrin-activated pathways including phosphoinositide 3-kinase, MAPK, JNK, and p38 nor did it influence activation of the integrin-responsive kinases focal adhesion kinase and Src. The identification of Trp(775) as a specific site for integrin-mediated Akt signaling supports the concept of specificity of signaling along the integrin cytoplasmic domain. PMID- 12637512 TI - Chromatin remodeling activities act on UV-damaged nucleosomes and modulate DNA damage accessibility to photolyase. AB - Nucleosomes inhibit DNA repair in vitro, suggesting that chromatin remodeling activities might be required for efficient repair in vivo. To investigate how structural and dynamic properties of nucleosomes affect damage recognition and processing, we investigated repair of UV lesions by photolyase on a nucleosome positioned at one end of a 226-bp-long DNA fragment. Repair was slow in the nucleosome but efficient outside. No disruption or movement of the nucleosome was observed after UV irradiation and during repair. However, incubation with the nucleosome remodeling complex SWI/SNF and ATP altered the conformation of nucleosomal DNA as judged by UV photo-footprinting and promoted more homogeneous repair. Incubation with yISW2 and ATP moved the nucleosome to a more central position, thereby altering the repair pattern. This is the first demonstration that two different chromatin remodeling complexes can act on UV-damaged nucleosomes and modulate repair. Similar activities might relieve the inhibitory effect of nucleosomes on DNA repair processes in living cells. PMID- 12637513 TI - Molecular cloning and characterization of CLICK-III/CaMKIgamma, a novel membrane anchored neuronal Ca2+/calmodulin-dependent protein kinase (CaMK). AB - During a screen for novel putative Ca(2+)/calmodulin-dependent protein kinase (CaMK)-like CREB kinases (CLICKs), we have cloned a full-length cDNA for CLICK III/CaMKIgamma, an isoform of the CaMKI family with an extended C-terminal domain ending with CAAX motif (where AA is aliphatic acid). As expected from the similarity of its kinase domain with the other CaMKI isoforms, full activation of CLICK-III/CaMKIgamma required both Ca(2+)/CaM and phosphorylation by CaMKK. We also found that Ca(2+)/cAMP-response element-binding protein (CREB) was a good substrate for CLICK-III/CaMKIgamma, at least in vitro. Interestingly enough, CLICK-III/CaMKIgamma transcripts were most abundant in neurons, with the highest levels in limited nuclei such as the central nucleus of the amygdala (CeA) and the ventromedial hypothalamus. Consistent with the presence of the CAAX motif, CLICK-III/CaMKIgamma was found to be anchored to various membrane compartments, especially to Golgi and plasma membranes. Both point mutation in the CAAX motif and treatment with compactin, a 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitor, disrupted such membrane localization, suggesting that membrane localization of CLICK-III/CaMKIgamma occurred in a prenylation-dependent way. These findings provide a novel mechanism by which neuronal CaMK activity could be targeted to specific membrane compartments. PMID- 12637514 TI - Oligomerization is a general mechanism for the activation of apoptosis initiator and inflammatory procaspases. AB - Proteolytic activation of initiator procaspases is a crucial step in the cellular commitment to apoptosis. Alternative models have been postulated for the activation mechanism, namely the oligomerization or induced proximity model and the allosteric regulation model. While the former holds that procaspases become activated upon proper oligomerization by an adaptor protein, the latter states that the adaptor is an allosteric regulator for procaspases. The allosteric regulation model has been applied for the activation of procaspase-9 by apoptotic protease-activating factor (Apaf-1) in an oligomeric complex known as the apoptosome. Using approaches that allow for controlled oligomerization, we show here that aggregation of multiple procaspase-9 molecules can induce their activation independent of the apoptosome. Oligomerization-induced procaspase-9 activation, both within the apoptosome and in artificial systems, requires stable homophilic association of the protease domains, raising the possibility that the function of Apaf-1 is not only to oligomerize procaspase-9 but also to maintain the interaction of the caspase-9 protease domain after processing. In addition, we provide biochemical evidence that other apoptosis initiator caspases (caspase 2 and -10) as well as a procaspase involved in inflammation (murine caspase-11) are also activated by oligomerization. Thus, oligomerization of precursor molecules appears to be a general mechanism for the activation of both apoptosis initiator and inflammatory procaspases. PMID- 12637515 TI - Physical and functional interaction of the yeast corepressor Tup1 with mRNA 5' triphosphatase. AB - The Tup1-Ssn6 complex is an important corepressor in Saccharomyces cerevisiae that inhibits transcription through interactions with the basal transcription machinery and by remodeling chromatin. In a two-hybrid screen for factors that interact with the Schizosaccharomyces pombe Tup1 ortholog, Tup11, we isolated the pct1+ cDNA. The pct1+ gene encodes an mRNA 5'-triphosphatase, which catalyzes the first step of mRNA capping reactions. Pct1 did not interact with the S. pombe Ssn6 ortholog. In vitro glutathione S-transferase pull-down experiments revealed that Pct1 binds to the WD repeat regions of Tup11 and the functionally redundant Tup12 protein. Similarly, the S. cerevisiae Tup1 protein associates with the mRNA 5'-triphosphatase encoded by the CET1 gene. The highly conserved C-terminal domain of Cet1 interacts with Tup1 in vitro, and Tup1-Ssn6 complexes co-purify with the Cet1 protein, indicating that in vivo interactions also occur between these proteins. Over-expression of CET1 compromised repression of an MFA2-lacZ reporter gene that is subject to Tup1-Ssn6 repression. These genetic and biochemical interactions between Tup1-Ssn6 and Cet1 indicate that the capping enzyme associated with RNA polymerase II is a target of the corepressor complex. PMID- 12637516 TI - Complex formation among the RNA export proteins Nup98, Rae1/Gle2, and TAP. AB - Most nucleocytoplasmic traffic through the nuclear pore complex is mediated by soluble receptors of the importin/exportin or karyopherin family. mRNA export is unique in that no receptor of this family has been implicated in trafficking of the bulk of mRNAs. Instead, many diverse proteins have been linked to mRNA export, but an all-encompassing model remains elusive. Understanding how these proteins interact with each other is central to the development of such a model. Here, we have focused on the interactions between three proteins implicated in mRNA export, Nup98, Rae1/Gle2, and TAP. We have defined the binary complexes that form among these proteins. We find that Gle2 requires two sites within TAP for stable interaction. Strikingly, rather than a general affinity for all nucleoporin FG repeats, TAP has highest affinity for a specific region within the GLFG domain of Nup98, indicating that not all repeats are identical in function. We have established that the ternary complex can form through simultaneous binding of both Gle2 and TAP to adjacent sites on Nup98. In contrast, Nup98 competes with TAP for Gle2 binding; when bound to Nup98, Gle2 no longer interacts directly with TAP. From these interactions, we propose that Gle2 may act to deliver TAP to Nup98 and that this may represent the first in a series of interactions between an export complex and a nucleoporin. PMID- 12637517 TI - RNA recognition motif-type RNA-binding proteins in Trypanosoma cruzi form a family involved in the interaction with specific transcripts in vivo. AB - Trypanosomes, protozoan parasites from the order Kinetoplastida, have to deal with environmental changes during the interaction with their hosts. Trypanosoma cruzi, the causative agent of Chagas' disease, uses post-transcriptional mechanisms to regulate gene expression. However, few RNA-binding proteins involved in mRNA turnover control have been identified to date. In this work, an RNA recognition motif (RRM)-type RNA-binding protein family named T. cruzi RNA binding protein (TcRBP) and composed of at least six members was identified. The genomic organization of four members revealed a head to tail arrangement within a region of 15 kilobase pairs. TcRBP members have a common RRM and different auxiliary domains with a high content of glycine, glutamine, and histidine residues within their N- and C-terminal regions. TcRBPs differ in their expression patterns as well as in their homoribopolymer binding interaction in vitro, although they preferentially recognize poly(U) and poly(G) RNAs. An interesting observation was the relaxed RNA-binding interactions with several trypanosome transcripts in vitro. In contrast, co-immunoprecipitation experiments of TcRBP-containing ribonucleoprotein complexes formed in vivo revealed a highly restricted binding interaction with specific RNAs. Several TcRBP-containing complexes are stage-specific and, in some cases, bear the poly(A)-binding protein TcPABP1. Altogether, these results suggest that TcRBPs might be modulated in vivo, to favor or preclude the interaction with specific transcripts in a developmentally regulated manner. PMID- 12637518 TI - The anomalous pKa of Tyr-9 in glutathione S-transferase A1-1 catalyzes product release. AB - The pKa of the catalytic Tyr-9 in glutathione S-transferase (GST) A1-1 is lowered from 10.3 to approximately 8.1 in the apoenzyme and approximately 9.0 with a GSH conjugate bound at the active site. However, a clear functional role for the unusual Tyr-9 pKa has not been elucidated. GSTA1-1 also includes a dynamic C terminus that undergoes a ligand-dependent disorder-to-order transition. Previous studies suggest a functional link between Tyr-9 ionization and C-terminal dynamics. Here we directly probe the role of Tyr-9 ionization in ligand binding and C-terminal conformation. An engineered mutant of rGSTA1-1, W21F/F222W, which contains a single Trp at the C terminus, was used as a fluorescent reporter of pH dependent C-terminal dynamics. This mutant exhibited a pH-dependent change in Trp 222 emission properties consistent with changes in C-terminal solvation or conformation. The apparent pKa values for the conformational transition were 7.9 +/- 0.1 and 9.3 +/- 0.1 for the apoenzyme and ligand-bound enzyme, respectively, in excellent agreement with the pKa for Tyr-9 in these states. The Y9F/W21F/F222W mutant, however, exhibited no such pH-dependent changes. Time-resolved fluorescence anisotropy studies revealed a ligand-dependent, Tyr-9-dependent, change in the order parameter of Trp-222. However, no pH dependence was observed. In equilibrium and pre-steady-state ligand binding studies, product conjugate had a decreased equilibrium binding affinity (KD), concomitant with increased binding and dissociation rates, at higher pH values. Furthermore, the recovered pKa values for the pH-dependent microscopic rate constants ranged from 7.7 to 8.4, also in agreement with the pKa of Tyr-9. In contrast, the Y9F/W21F/F222W mutant had no pH-dependent transition in KD or rate constants for ligand binding or dissociation. The combined results indicate that the macroscopic populations of "open" and "closed" states of the C terminus are not determined solely by the ionization state of Tyr-9. However, the rates of transition between these states are faster for the ionized Tyr-9. The ionized Tyr-9 states provide a parallel pathway for product dissociation, which is kinetically and thermodynamically favored. In silico kinetic models further support the functional role for the parallel dissociation pathway provided by ionized Tyr-9. PMID- 12637519 TI - Crystal structure of a bacterial endospore coat component. A laccase with enhanced thermostability properties. AB - Endospores produced by the Gram-positive soil bacterium Bacillus subtilis are shielded by a proteinaceous coat formed by over 30 structural components, which self-assemble into a lamellar inner coat and a thicker striated electrodense outer coat. The 65-kDa CotA protein is an abundant component of the outer coat layer. CotA is a highly thermostable laccase, assembly of which into the coat is required for spore resistance against hydrogen peroxide and UV light. Here, we report the structure of CotA at 1.7-A resolution, as determined by x-ray crystallography. This is the first structure of an endospore coat component, and also the first structure of a bacterial laccase. The overall fold of CotA comprises three cupredoxin-like domains and includes one mononuclear and one trinuclear copper center. This arrangement is similar to that of other multicopper oxidases and most similar to that of the copper tolerance protein CueO of Escherichia coli. However, the three cupredoxin domains in CotA are further linked by external interdomain loops, which increase the packing level of the structure. We propose that these interdomain loops contribute to the remarkable thermostability of the enzyme, but our results suggest that additional factors are likely to play a role. Comparisons with the structure of other monomeric multicopper oxidases containing four copper atoms suggest that CotA may accept the largest substrates of any known laccase. Moreover, and unlike other laccases, CotA appears to have a flexible lidlike region close to the substrate binding site that may mediate substrate accessibility. The implications of these findings for the properties of CotA, its assembly and the properties of the bacterial spore coat structure are discussed. PMID- 12637520 TI - NTP-driven translocation by human RNA polymerase II. AB - We report a "running start, two-bond" protocol to analyze elongation by human RNA polymerase II (RNAP II). In this procedure, the running start allowed us to measure rapid rates of elongation and provided detailed insight into the RNAP II mechanism. Formation of two bonds was tracked to ensure that at least one translocation event was analyzed. By using this method, RNAP II is stalled briefly at a defined template position before restoring the next NTP. Significantly, slow reaction steps are identified both before and after phosphodiester bond synthesis, and both of these steps can be highly dependent on the next templated NTP. The initial and final NTP-driven events, however, are not identical, because the slow step after chemistry, which includes translocation and pyrophosphate release, is regulated differently by elongation factors hepatitis delta antigen and transcription factor IIF. Because recovery from a stall and the processive transition from one bond to the next can be highly NTP dependent, we conclude that translocation can be driven by the incoming substrate NTP, a model fully consistent with the RNAP II elongation complex structure. PMID- 12637522 TI - Multifunctional roles for the PH domain of Dbs in regulating Rho GTPase activation. AB - Dbl family members are guanine nucleotide exchange factors specific for Rho guanosine triphosphatases (GTPases) and invariably possess tandem Dbl (DH) and pleckstrin homology (PH) domains. Dbs, a Dbl family member specific for Cdc42 and RhoA, exhibits transforming activity when overexpressed in NIH 3T3 mouse fibroblasts. In this study, the PH domain of Dbs was mutated to impair selectively either guanine nucleotide exchange or phosphoinositide binding in vitro and resulting physiological alterations were assessed. As anticipated, substitution of residues within the PH domain of Dbs integral to the interface with GTPases reduced nucleotide exchange and eliminated the ability of Dbs to transform NIH 3T3 cells. More interestingly, substitutions within the PH domain that prevent interaction with phosphoinositides yet do not alter in vitro activation of GTPases also do not transform NIH 3T3 cell and fail to activate RhoA in vivo despite proper subcellular localization. Therefore, the PH domain of Dbs serves multiple roles in the activation of GTPases and cannot be viewed as a simple membrane-anchoring device. In particular, the data suggest that binding of phosphoinositides to the PH domain within the context of membrane surfaces may direct orientations or conformations of the linked DH and PH domains to regulate GTPases activation. PMID- 12637521 TI - Two distinct phases of virus-induced nuclear factor kappa B regulation enhance tumor necrosis factor-related apoptosis-inducing ligand-mediated apoptosis in virus-infected cells. AB - Cellular transcription factors are often utilized by infecting viruses to promote viral growth and influence cell fate. We have previously shown that nuclear factor kappaB (NF-kappaB) is activated after reovirus infection and that this activation is required for virus-induced apoptosis. In this report we identify a second phase of reovirus-induced NF-kappaB regulation. We show that at later times post-infection NF-kappaB activation is blocked in reovirus-infected cells. This results in the termination of virus-induced NF-kappaB activity and the inhibition of tumor necrosis factor alpha and etoposide-induced NF-kappaB activation in infected cells. Reovirus-induced inhibition of NF-kappaB activation occurs by a mechanism that prevents IkappaBalpha degradation and that is blocked in the presence of the viral RNA synthesis inhibitor, ribavirin. Reovirus-induced apoptosis is mediated by tumor necrosis factor-related apoptosis inducing ligand (TRAIL) in a variety of epithelial cell lines. Herein we show that ribavirin inhibits reovirus-induced apoptosis in TRAIL-resistant HEK293 cells and prevents the ability of reovirus infection to sensitize TRAIL-resistant cells to TRAIL induced apoptosis. Furthermore, TRAIL-induced apoptosis is enhanced in HEK293 cells expressing IkappaBDeltaN2, which blocks NF-kappaB activation. These results indicate that the ability of reovirus to inhibit NF-kappaB activation sensitizes HEK293 cells to TRAIL and facilitates virus-induced apoptosis in TRAIL-resistant cells. Our findings demonstrate that two distinct phases of virus-induced NF kappaB regulation are required to efficiently activate host cell apoptotic responses to reovirus infection. PMID- 12637523 TI - Identification of a novel nuclear localization signal common to 69- and 82-kDa human choline acetyltransferase. AB - We demonstrated previously that 69- and 82-kDa human choline acetyltransferase are localized predominantly to the cytoplasm and the nucleus, respectively. We have now identified a nuclear localization signal common to both forms of enzyme using confocal microscopy to study the subcellular compartmentalization of choline acetyltransferase tagged with green fluorescent protein in living HEK 293 cells. To identify functional nuclear localization and export signals, portions of full-length 69-kDa choline acetyltransferase were cloned into the vector peGFP N1 and the cellular distribution patterns of the fusion proteins observed. Of the nine constructs studied, one yielded a protein with nuclear localization and another produced a protein with cytoplasmic localization. Mutation of the critical amino acids in this novel putative nuclear localization signal in the 69 and 82-kDa enzymes demonstrated that it is functional in both proteins. Moreover, 69-kDa choline acetyltransferase but not the 82-kDa enzyme is transported out of the nucleus by the leptomycin B-sensitive Crm-1 export pathway. By using bikaryon cells expressing both 82-kDa choline acetyltransferase and the nuclear protein heterogeneous nuclear ribonucleoprotein with green and red fluorescent tags, respectively, we found that the 82-kDa enzyme does not shuttle out of the nucleus in measurable amounts. These data suggest that 69-kDa choline acetyltransferase is a nucleocytoplasmic shuttling protein with a predominantly cytoplasmic localization determined by a functional nuclear localization signal and unidentified putative nuclear export signal. For 82-kDa choline acetyltransferase, the presence of the unique amino-terminal nuclear localization signal plus the newly identified nuclear localization signal may be involved in a process leading to predominantly nuclear accumulation of this enzyme, or alternatively, the two nuclear localization signals may be sufficient to overcome the force(s) driving nuclear export. PMID- 12637524 TI - Zinc finger protein Prz1 regulates Ca2+ but not Cl- homeostasis in fission yeast. Identification of distinct branches of calcineurin signaling pathway in fission yeast. AB - Calcineurin is an important mediator that connects the Ca(2+)-dependent signaling to various cellular responses in a wide variety of cell types and organisms. In budding yeast, activated calcineurin exerts its function mainly by regulating the Crz1p/Tcn1 transcription factor. Here, we cloned the fission yeast prz1(+) gene, which encodes a zinc finger transcription factor highly homologous to Crz1/Tcn1. Similar to the results in budding yeast, calcineurin dephosphorylated Prz1 and resulted in the trans-location of Prz1 from the cytoplasm to the nucleus. Prz1 expression was stimulated by high extracellular Ca(2+) in a calcineurin-dependent fashion. However, unlike in budding yeast, the prz1-null cells did not show any phenotype similar to those previously reported in calcineurin deletion such as aberrant cell morphology, mating defect, or hypersensitivity to Cl(-). Instead, the prz1-null cells showed hypersensitivity to Ca(2+), consistent with a dramatic decrease in transcription of Pmc1 Ca(2+) pump. Interestingly, overexpression of Prz1 did not suppress the Cl(-) hypersensitivity of calcineurin deletion, and overexpression of Pmp1 MAPK phosphatase suppressed the Cl(-) hypersensitivity of calcineurin deletion but not the Ca(2+) hypersensitivity of prz1 deletion. In addition, mutations in the its2(+)/cps1(+), its8(+), and its10(+)/cdc7(+) genes that showed synthetic lethal genetic interaction with calcineurin deletion did not exhibit synthetic lethality with the prz1 deletion. Our results suggest that calcineurin activates at least two distinct signaling branches, i.e. the Prz1 dependent transcriptional regulation and an unknown mechanism, which functions antagonistically with the Pmk1 MAPK pathway. PMID- 12637525 TI - CCAAT/enhancer-binding protein and activator protein-1 transcription factors regulate the expression of interleukin-8 through the mitogen-activated protein kinase pathways in response to mechanical stretch of human airway smooth muscle cells. AB - Here we investigated the mechanisms by which mechanical stretch regulates the production of IL-8 in primary human airway smooth muscle cells (HASMC). Bronchial HASMC were subjected to cyclic mechanical stretch (12%, 1 Hz) using the computer controlled Flexcell Strain system. Mechanical stretch increased IL-8 mRNA expression and protein production. Cyclic stretch of HASMC also increased the kinase activities of ERK1/2, JNK1, p38, and the DNA binding activities of AP-1 and C/EBP transcription factors with little effect on NF-kappa B. The inhibition of AP-1 and C/EBP transcriptional activities blocked the production of IL-8 in culture supernatants. Furthermore, the inhibition of ERK1/2 and p38 but not JNK1 caused a significant down-regulation in the expression and production of IL-8 in response to cyclic stretch. Although protein tyrosine kinases were required for the activation of both ERK1/2 and p38 kinase, stretch-activated channels, small GTPase proteins, and extracellular Ca2+ influx were required only for the activation of p38 kinase whereas phosphoinositide 3-kinase was needed for ERK1/2 activation. In addition, the phosphorylation of ERK1/2 was essential for the activation of AP-1 whereas p38 MAP kinase was needed for the activation of C/EBP. Our data demonstrate that the cyclic stretch of HASMC causes the increased production of IL-8 by activating the AP-1 and C/EBP transcription factors through the activation of ERK1/2 and p38 kinase signaling pathways. PMID- 12637526 TI - Characterization of the MRP4- and MRP5-mediated transport of cyclic nucleotides from intact cells. AB - Cyclic nucleotides are known to be effluxed from cultured cells or isolated tissues. Two recently described members of the multidrug resistance protein family, MRP4 and MRP5, might be involved in this process, because they transport the 3',5'-cyclic nucleotides, cAMP and cGMP, into inside-out membrane vesicles. We have investigated cGMP and cAMP efflux from intact HEK293 cells overexpressing MRP4 or MRP5. The intracellular production of cGMP and cAMP was stimulated with the nitric oxide releasing compound sodium nitroprusside and the adenylate cyclase stimulator forskolin, respectively. MRP4- and MRP5-overexpressing cells effluxed more cGMP and cAMP than parental cells in an ATP-dependent manner. In contrast to a previous report we found no glutathione requirement for cyclic nucleotide transport. Transport increased proportionally with intracellular cyclic nucleotide concentrations over a calculated range of 20-600 microm, indicating low affinity transport. In addition to several classic inhibitors of organic anion transport, prostaglandins A(1) and E(1), the steroid progesterone and the anti-cancer drug estramustine all inhibited cyclic nucleotide efflux. The efflux mediated by MRP4 and MRP5 did not lead to a proportional decrease in the intracellular cGMP or cAMP levels but reduced cGMP by maximally 2-fold over the first hour. This was also the case when phosphodiesterase-mediated cyclic nucleotide hydrolysis was inhibited by 3-isobutyl-1-methylxanthine, conditions in which efflux was maximal. These data indicate that MRP4 and MRP5 are low affinity cyclic nucleotide transporters that may at best function as overflow pumps, decreasing steep increases in cGMP levels under conditions where cGMP synthesis is strongly induced and phosphodiesterase activity is limiting. PMID- 12637527 TI - c-Myc directly regulates the transcription of the NBS1 gene involved in DNA double-strand break repair. AB - The c-myc proto-oncogene encodes a ubiquitous transcription factor involved in the control of cell growth and implicated in inducing tumorigenesis. Understanding the function of c-Myc and its role in cancer depends upon the identification of c-Myc target genes. Nijmegen breakage syndrome (NBS) is a chromosomal-instability syndrome associated with cancer predisposition, radiosensitivity, and chromosomal instability. The NBS gene product, NBS1 (p95 or nibrin), is a part of the hMre11 complex, a central player associated with double strand break (DSB) repair. NBS1 contains domains characteristic for proteins involved in DNA repair, recombination, and replication. Here we show that c-Myc directly activates NBS1. c-Myc-mediated induction of NBS1 gene transcription occurs in different tissues, is independent of cell proliferation, and is mediated by a c-Myc binding site in the intron 1 region of NBS1 gene. Overexpression of NBS1 in Rat1a cells increased cell proliferation. These results indicate that NBS1 is a direct transcriptional target of c-Myc and links the function of c-Myc to the regulation of DNA DSB repair pathway operating during DNA replication. PMID- 12637528 TI - Smad3 potentiates transforming growth factor beta (TGFbeta )-induced apoptosis and expression of the BH3-only protein Bim in WEHI 231 B lymphocytes. AB - Transforming growth factor-beta (TGFbeta) is a potent growth inhibitor and inducer of apoptosis in B lymphocytes and is essential for immune regulation and maintenance of self-tolerance. Here we show that exogenous overexpression of Smad3 potentiates TGFbeta-induced apoptosis and expression of the pro-apoptotic protein Bim in WEHI 231 B lymphocytes. Overexpression of dominant-negative forms of Smad3 abrogate these TGFbeta-induced responses. We also demonstrate that TGFbeta induces Bim protein expression concomitant with its induction of apoptosis in the mouse progenitor B lymphocyte cell line, Ba/F3. Enhanced expression of Bim protein induced by TGFbeta is associated with an increased association of Bim with Bcl-2 and a concomitant loss of mitochondrial membrane potential. Furthermore, we find that the anti-apoptotic effect of the pro survival cytokine CD40 results in the abrogation of TGFbeta-mediated Bim induction. Our data provide the first evidence of Bim expression levels that are increased by the addition of a pro-apoptotic cytokine, TGFbeta, and also suggest that the TGFbeta-specific transcription factor Smad3 plays a role in mediating Bim expression levels and apoptosis. PMID- 12637529 TI - Direct detection of Fe(IV)[double bond]O intermediates in the cytochrome aa3 oxidase from Paracoccus denitrificans/H2O2 reaction. AB - We report the first evidence for the formation of the "607- and 580-nm forms" in the cytochrome oxidase aa3/H2O2 reaction without the involvement of tyrosine 280. The pKa of the 607-580-nm transition is 7.5. The 607-nm form is also formed in the mixed valence cytochrome oxidase/O2 reaction in the absence of tyrosine 280. Steady-state resonance Raman characterization of the reaction products of both the wild-type and Y280H cytochrome aa3 from Paracoccus denitrificans indicate the formation of six-coordinate low spin species, and do not support, in contrast to previous reports, the formation of a porphyrin pi-cation radical. We observe three oxygen isotope-sensitive Raman bands in the oxidized wild-type aa3/H2O2 reaction at 804, 790, and 358 cm-1. The former two are assigned to the Fe(IV)[double bond]O stretching mode of the 607- and 580-nm forms, respectively. The 14 cm-1 frequency difference between the oxoferryl species is attributed to variations in the basicity of the proximal to heme a3 His-411, induced by the oxoferryl conformations of the heme a3-CuB pocket during the 607-580-nm transition. We suggest that the 804-790 cm-1 oxoferryl transition triggers distal conformational changes that are subsequently communicated to the proximal His-411 heme a3 site. The 358 cm-1 mode has been found for the first time to accumulate with the 804 cm-1 mode in the peroxide reaction. These results indicate that the mechanism of oxygen reduction must be reexamined. PMID- 12637530 TI - Critical role of the pleckstrin homology domain in Dbs signaling and growth regulation. AB - Dbl family proteins act as guanine nucleotide exchange factors and positive regulators of Rho GTPase function by stimulating formation of the active, GTP bound state. All Dbl family Rho guanine nucleotide exchange factors possess an invariant tandem domain structure consisting of a Dbl homology (DH) catalytic domain followed by a pleckstrin homology (PH) regulatory domain. We determined previously that the PH domain of Dbs was critical for the intrinsic catalytic activity of the DH domain in vitro and for Dbs transformation in vivo. In this study, we evaluated the role of phosphoinositide binding to the PH domain in regulating the DH domain function of Dbs in vitro and in vivo. We determined that mutation of basic amino acids located within the beta1-beta2 and beta3-beta4 loops of the PH domain resulted in impaired phospholipid binding in vitro, yet full guanine nucleotide exchange activity in vitro was retained for RhoA and Cdc42. Surprisingly, these mutants were compromised in their ability to activate Rho GTPases in vivo and to cause transformation of NIH 3T3 cells. However, Dbs subcellular localization was impaired by these PH domain mutations, supporting a role for phospholipid interactions in facilitating membrane association. Despite the importance of phospholipid binding for Dbs function in vivo, we found that Dbs signaling and transforming activity was not stimulated by phosphatidylinositol 3-kinase activation. We suggest that the PH domain of Dbs facilitates two distinct roles in the regulation of DH domain function, one critical for GTPase association and activation in vitro and one critical for phosphoinositide binding and GTPase interaction in vivo, that together promote Dbs association with membranes. PMID- 12637531 TI - Phosphorylation-dependent and -independent nuclear import of RS domain-containing splicing factors and regulators. AB - SR proteins and related RS domain-containing polypeptides are an important class of splicing regulators in higher eukaryotic cells. The RS domain facilitates nuclear import of SR proteins and mediates protein-protein interactions during spliceosome assembly; both functions appear to subject to regulation by phosphorylation. Previous studies have identified two nuclear import receptors for SR proteins, transportin-SR1 and transportin-SR2. Here we show that transportin-SR1 and transportin-SR2 are the alternatively spliced products of the same gene and that transportin-SR2 is the predominant transcript in most cells and tissues examined. While both receptors import typical SR proteins in a phosphorylation-dependent manner, they differentially import the RS domain containing splicing regulators hTra2alpha and hTra2beta in different phosphorylation states. We suggest that differential regulation of nuclear import may serve as a mechanism for homeostasis of RS domain-containing splicing factors and regulators in the nucleus and for selective cellular responses to signaling. PMID- 12637532 TI - Activation of the programmed cell death pathway by inhibition of proteasome function in plants. AB - Proteasomes constitute the major machinery to degrade or process proteins by ATP/ubiquitin-mediated proteolysis. Recent findings suggest a pivotal role of the ubiquitin/proteasome pathway in the regulation of apoptosis in animal cells. Here we show that virus-induced gene silencing of two different subunits of the 26 S proteasome, the alpha 6 subunit of the 20 S proteasome and RPN9 subunit of 19 S regulatory complex, both activated the programmed cell death (PCD) program, accompanied by reduced proteasome activity and accumulation of polyubiquitinated proteins. These results demonstrate that disruption of proteasome function leads to PCD in plant cells. The affected cells showed morphological markers of PCD, including nuclear condensation and DNA fragmentation, accompanied by the 10-fold higher production of reactive oxygen species and increased ion leakage for 3 fold. Similar to apoptosis in animal system, mitochondrial membrane potential was decreased, cytochrome c released from mitochondria to cytosol, and caspase 9- and caspase 3-like proteolytic activities detected in the cells. Interestingly, this proteasome-mediated PCD stimulated the expression of only a subset of transcripts that are highly induced during pathogen-mediated hypersensitive response cell death, indicating that the two PCD pathways are differentially regulated. Taken together, these results provide the first direct evidence that proteasomes play a role in the regulatory program of PCD in plants. Controlled inhibition of proteasome activities may be involved in developmentally or environmentally activated plant cell death programs. PMID- 12637534 TI - Crystal structure of yeast cytosine deaminase. Insights into enzyme mechanism and evolution. AB - Yeast cytosine deaminase is an attractive candidate for anticancer gene therapy because it catalyzes the deamination of the prodrug 5-fluorocytosine to form 5 fluorouracil. We report here the crystal structure of the enzyme in complex with the inhibitor 2-hydroxypyrimidine at 1.6-A resolution. The protein forms a tightly packed dimer with an extensive interface of 1450 A2 per monomer. The inhibitor was converted into a hydrated adduct as a transition-state analog. The essential zinc ion is ligated by the 4-hydroxyl group of the inhibitor together with His62, Cys91, and Cys94 from the protein. The enzyme shares similar active site architecture to cytidine deaminases and an unusually high structural homology to 5-aminoimidazole-4-carboxamide-ribonucleotide transformylase and thereby may define a new superfamily. The unique C-terminal tail is involved in substrate specificity and also functions as a gate controlling access to the active site. The complex structure reveals a closed conformation, suggesting that substrate binding seals the active-site entrance so that the catalytic groups are sequestered from solvent. A comparison of the crystal structures of the bacterial and fungal cytosine deaminases provides an elegant example of convergent evolution, where starting from unrelated ancestral proteins, the same metal assisted deamination is achieved through opposite chiral intermediates within distinctly different active sites. PMID- 12637533 TI - Multiple mechanisms of reduced major histocompatibility complex class II expression in endotoxin tolerance. AB - Patients after polytrauma, burns, or septic shock frequently develop a life threatening immunodeficiency. This state is associated with specific functional alterations of monocytic cells. We previously proposed endotoxin tolerance, the monocyte state after acute response to lipopolysaccharide, as a respective model system. One major feature in both the clinical situation and the in vitro model is the dramatic down-regulation of monocyte major histocompatibility complex (MHC) class II surface expression, which is associated with impaired antigen presentation capacity. This study focused on the mechanisms behind reduced MHC class II expression in endotoxin tolerance. Endotoxin priming provoked a decrease of monocyte intracellular MHC class II. It also led to a reduced expression of the chaperonic invariant chain and to an inhibited synthesis of the major lysosomal enzyme for final cleavage of the invariant chain going along with a relative accumulation of p10. The expression of HLA-DM necessary for loading MHC class II with antigenic peptide was also decreased. Additionally, reduced export of MHC class II alphabeta complexes to the cell surface was observed. The down regulation of HLA-DR, invariant chain, and HLA-DM was regulated at the mRNA level and may be the consequence of reduced class II transactivator expression observed in this study. The simultaneous interference at different regulatory levels may explain the uniquely strong and long lasting MHC class II down-modulating effect of endotoxin priming compared with transforming growth factor-beta and interleukin-10. These results not only contribute to a better understanding of experimental endotoxin tolerance but may also give rise to new therapeutics for temporary immunodeficiency and, conversely, for MHC class II-dependent diseases such as autoimmunity and transplant rejection. PMID- 12637535 TI - Structure and intermolecular interactions of the luminal dimerization domain of human IRE1alpha. AB - Accumulation of unfolded proteins in the lumen of the endoplasmic reticulum activates a signal transduction cascade that culminates in the transcriptional induction of genes encoding adaptive functions. One proximal sensor for this unfolded protein response is the protein kinase/endoribonuclease IRE1alpha. IRE1alpha is a type-I transmembrane glycoprotein for which the N-terminal luminal domain (NLD) senses the accumulation of unfolded proteins. Previously we demonstrated that the NLD forms a stable ligand-independent dimer linked by disulfide bridges. In this report we have identified the cysteine residues responsible for intermolecular disulfide bonding. However, this covalent interaction was not required for dimerization and/or signaling, suggesting that a cryptic dimer interface exists in the NLD that is independent of covalent disulfide interactions. Limited proteolysis of the NLD revealed characteristic fragments, all retaining the same N-terminal sequences as full-length NLD. Biochemical and functional studies using NLD truncation mutants indicated that the dimerization domain of the NLD is confined to the conserved motifs at the N terminal regions where putative hydrophobic interactions exist. In addition, the peptide binding domain of the endoplasmic reticulum protein chaperone BiP interacted with the N-terminal region within the NLD. Our findings suggest that the NLD has at least two distinct types of interactions mediating dimerization and function in signaling, i.e. covalent interactions involving disulfide bond formation and hydrophobic interactions, with the hydrophobic interaction being the driving force for dimerization. PMID- 12637536 TI - A Novel H-NS-like protein from an antarctic psychrophilic bacterium reveals a crucial role for the N-terminal domain in thermal stability. AB - We describe here new members of the H-NS protein family identified in a psychrotrophic Acinetobacter spp. bacterium collected in Siberia and in a psychrophilic Psychrobacter spp. bacterium collected in Antarctica. Both are phylogenetically closely related to the HvrA and SPB Rhodobacter transcriptional regulators. Their amino acid sequence shares 40% identity, and their predicted secondary structure displays a structural and functional organization in two modules similar to that of H-NS in Escherichia coli. Remarkably, the Acinetobacter protein fully restores to the wild-type H-NS-dependent phenotypes, whereas the Psychrobacter protein is no longer able to reverse the effects of H NS deficiency in an E. coli mutant strain above 30 degrees C. Moreover, in vitro experiments demonstrate that the ability of the Psychrobacter H-NS protein to bind curved DNA and to form dimers is altered at 37 degrees C. The construction of hybrid proteins containing the N- or the C-terminal part of E. coli H-NS fused to the C- or N-terminal part of the Psychrobacter protein demonstrates the role of the N-terminal domain in this process. Finally, circular dichroism analysis of purified H-NS proteins suggests that, as compared with the E. coli and Acinetobacter proteins, the alpha-helical domain displays weaker intermolecular interactions in the Psychrobacter protein, which may account for the low thermal stability observed at 37 degrees C. PMID- 12637537 TI - Bone morphogenetic protein-1 (BMP-1). Identification of the minimal domain structure for procollagen C-proteinase activity. AB - Bone morphogenetic protein-1 (BMP-1) is a shorter spliced variant of mammalian tolloid (mTld), both of which cleave the C-propeptides of type I procollagen during the synthesis of extracellular matrix collagen fibrils. The fact that BMP 1 and mTld both exhibit procollagen C-proteinase (PCP) activity and that BMP-1 is the smaller variant might indicate that BMP-1 comprises the minimal required sequences for PCP activity. BMP-1 comprises a metalloproteinase domain, three CUB domains, and an epidermal growth factor (EGF)-like domain, which is located between the second and third CUB (complement components C1r/C1s, the sea urchin protein Uegf, and BMP-1) domains. In this study we showed the following. 1) The CUB1 domain is required for secretion of the molecule. Domain swapping experiments, in which CUB1 and other CUB domains were interchanged, resulted in retention of the proteins by cells. Therefore, CUB1 and its location immediately adjacent to the metalloproteinase domain are essential for secretion of the protein. 2) Mutants lacking the EGF-like and CUB3 domains exhibited full C proteinase activity. In contrast, mutants lacking the CUB2 domain were poor C proteinases. 3) Further studies showed that Glu-483 on the beta4-beta5 loop of CUB2 is essential for C-proteinase activity of BMP-1. In conclusion, the study showed that the minimal domain structure for PCP activity is considerably shorter than expected and comprises the metalloproteinase domain and the CUB1 and CUB2 domains of BMP-1. PMID- 12637538 TI - Tyrosine phosphorylation of protein kinase D in the pleckstrin homology domain leads to activation. AB - Protein kinase D (PKD) is a member of the AGC family of Ser/Thr kinases and is distantly related to protein kinase C (PKC). Formerly known as PKCmu, PKD contains protein domains not found in conventional PKC isoforms. A functional pleckstrin homology (PH) domain is critical for the regulation of PKD activity. Here we report that PKD is tyrosine-phosphorylated within the PH domain, leading to activation. This phosphorylation is mediated by a pathway that consists of the Src and Abl tyrosine kinases and occurs in response to stimulation with pervanadate and oxidative stress. Mutational analysis revealed three tyrosine phosphorylation sites (Tyr(432), Tyr(463), and Tyr(502)), which are regulated by the Src-Abl pathway, and phosphorylation of only one of these (Tyr(463)) leads to PKD activation. By using a phospho-specific antibody, we show that Abl directly phosphorylates PKD at Tyr(463) in vitro, and in cells phosphorylation of this site is sufficient to mediate full activation of PKD. Mutation of the other two sites, Tyr(432) and Tyr(502), had no significant influence on PKD activity. These data reveal a tyrosine phosphorylation-dependent activation mechanism for PKD and suggest that this event contributes to the release of the autoinhibitory PKD PH domain leading to kinase activation and downstream responses. PMID- 12637539 TI - D-Peptides as inhibitors of the DnaK/DnaJ/GrpE chaperone system. AB - DnaK, a Hsp70 homolog of Escherichia coli, together with its co-chaperones DnaJ and GrpE protects denatured proteins from aggregation and promotes their refolding by an ATP-consuming mechanism. DnaJ not only stimulates the gamma phosphate cleavage of DnaK-bound ATP but also binds polypeptide substrates on its own. Unfolded polypeptides, such as denatured luciferase, thus form ternary complexes with DnaJ and DnaK. A previous study has shown that d-peptides compete with l-peptides for the same binding site in DnaJ but do not bind to DnaK (Feifel, B., Schonfeld, H.-J., and Christen, P. (1998) J. Biol. Chem. 273, 11999 12002). Here we report that d-peptides efficiently inhibit the refolding of denatured luciferase by the DnaK/DnaJ/GrpE chaperone system (EC50 = 1-2 microM). The inhibition of the chaperone action is due to the binding of d-peptide to DnaJ (Kd = 1-2 microM), which seems to preclude DnaJ from forming ternary (ATP.DnaK)m.substrate.DnaJn complexes. Apparently, simultaneous binding of DnaJ and DnaK to one and the same target polypeptide is essential for effective chaperone action. PMID- 12637540 TI - The role of transcription initiation factor IIIB subunits in promoter opening probed by photochemical cross-linking. AB - The core transcription initiation factor (TF) IIIB recruits its conjugate RNA polymerase (pol) III to the promoter and also plays an essential role in promoter opening. TFIIIB assembled with certain deletion mutants of its Brf1 and Bdp1 subunits is competent in pol III recruitment, but the resulting preinitiation complex does not open the promoter. Whether Brf1 and Bdp1 participate in opening the promoter by direct DNA interaction (as sigma subunits of bacterial RNA polymerases do) or indirectly by their action on pol III has been approached by site-specific photochemical protein-DNA cross-linking of TFIIIB-pol III-U6 RNA gene promoter complexes. Brf1, Bdp1, and several pol III subunits can be cross linked to the nontranscribed strand of the U6 promoter at base pair -9/-8 and +2/+3 (relative to the transcriptional start as +1), respectively the upstream and downstream ends of the DNA segment that opens up into the transcription bubble. Cross-linking of Bdp1 and Brf1 is detected at 0 degrees C in closed preinitiation complexes and at 30 degrees C in complexes that are partly open, but also it is detected in mutant TFIIIB-pol III-DNA complexes that are unable to open the promoter. In contrast, promoter opening-defective TFIIIB mutants generate significant changes of cross-linking of polymerase subunits. The weight of this evidence argues in favor of an indirect mode of action of TFIIIB in promoter opening. PMID- 12637541 TI - A two-component system mediates developmental regulation of biosynthesis of a heterocyst polysaccharide. AB - Some cyanobacteria couple oxygenic photosynthesis in vegetative cells with O2 sensitive N2 fixation in differentiated cells called heterocysts. Heterocyst differentiation involves extensive biochemical and structural changes that collectively permit heterocysts to assimilate N2 aerobically and supply the products of N2 fixation to vegetative cells. HepK and DevR are required for the development of functional heterocysts in Anabaena and Nostoc, respectively. We show that HepK is an autokinase and that Anabaena DevRA is its cognate response regulator, together comprising part or all of a two-component system that mediates developmental regulation of biosynthesis of a heterocyst envelope polysaccharide. Recombinant N-hexahistidine-tagged HepK (H6HepK), the cytoplasmic portion H6'HepK of H6HepK, H6DevR, and H6DevRA were overexpressed in Escherichia coli and purified to homogeneity. H6'HepK, but not H6HepK, autophosphorylates with [gamma-32P]ATP. ADP, specifically, elicits dephosphorylation of phosphorylated H6'HepK. The phosphoryl group of H6'HepK is transferred rapidly and efficiently to both H6DevR and H6DevRA but not to His-tagged OmpR, whose cognate sensor kinase is EnvZ. Sequence comparisons, the results of site-specific mutagenesis, and tests of chemical stability support identification of HepK His348 and DevR-Asp53 as the phosphorylated residues. The mutation HepK-H348A abolishes both in vitro autokinase activity and in vivo functionality of HepK. Heterocysts of both hepK Anabaena and devRA Anabaena lack an envelope polysaccharide layer and are nonfunctional. Consistent with the normal site of deposition of that polysaccharide, a hepK::gfp transcriptional fusion is expressed principally in proheterocysts. HepK/DevRA is the first two-component system identified that regulates the biosynthesis of a polysaccharide as part of a patterned differentiation process. PMID- 12637542 TI - Complex transcription and splicing of odorant receptor genes. AB - Human major histocompatibility (human leucocyte antigen (HLA)) complex-linked odorant receptor (OR) genes are among the best characterized OR genes in the human genome. In addition to their functions as odorant receptors in olfactory epithelium, they have been suggested to play a role in the fertilization process. Here, we report the first in-depth analysis of their expression and regulation within testicular tissue. Sixteen HLA-linked OR and three non-HLA-linked OR were analyzed. One OR gene (hs6M1-16, in positive transcriptional orientation) exhibited six different transcriptional start sites combined with extensive alternative splicing within the 5'-untranslated region, the coding exon, and the 3'-untranslated region. Long distance splicing, exon sharing, and premature polyadenylation were features of another three OR loci (hs6M1-18, -21, and -27, all upstream of hs6M1-16, but in negative transcriptional orientation). Determination of the transcriptional start sites of these OR genes identified a region of 81 bp with potential bi-directional transcriptional activity. The results demonstrate that HLA-linked OR genes are subject to unusually complex transcriptional regulatory mechanisms. PMID- 12637543 TI - Identification of novel domains within Sox-2 and Sox-11 involved in autoinhibition of DNA binding and partnership specificity. AB - Sox transcription factors play key regulatory roles throughout development, binding DNA through a consensus (A/T)(A/T)CAA(A/T)G sequence. Although many different Sox proteins bind to this sequence, it has been observed that gene regulatory elements are commonly responsive to only a small subset of the entire family, implying that regulatory mechanisms exist to permit selective DNA binding and/or transactivation by Sox family members. To identify and explore the mechanisms modulating gene activation by Sox proteins further, we compared the function of Sox-2 and Sox-11. This led to the discovery that Sox proteins are regulated differentially at multiple levels, including transactivation, protein partnerships with Pit-Oct-Unc (POU) transcription factors, and DNA binding autoregulation. Specifically, we determined that Sox-11 activates transcription more strongly than Sox-2 and that the transactivation domain of Sox-11 is primarily responsible for this capability. Additionally, we demonstrate that the Sox-11 DNA binding domain is responsible for selective cooperation with the POU factor Brn-2. This requirement cannot be replaced by the DNA binding domain of Sox-2, indicating that the DNA binding domain of Sox proteins is critical for Sox POU partnerships. Interestingly, we have also determined that a conserved domain of Sox-11 has the novel capability of autoinhibiting its ability to bind DNA in vitro and to activate gene expression in vivo. Our findings suggest that the autoinhibitory domain can repress promiscuous binding of Sox-11 to DNA and plays an important role in regulating the recruitment of Sox-11 to specific genes. PMID- 12637544 TI - Biochemical and molecular characterization of a hydroxyjasmonate sulfotransferase from Arabidopsis thaliana. AB - 12-Hydroxyjasmonate, also known as tuberonic acid, was first isolated from Solanum tuberosum and was shown to have tuber-inducing properties. It is derived from the ubiquitously occurring jasmonic acid, an important signaling molecule mediating diverse developmental processes and plant defense responses. We report here that the gene AtST2a from Arabidopsis thaliana encodes a hydroxyjasmonate sulfotransferase. The recombinant AtST2a protein was found to exhibit strict specificity for 11- and 12-hydroxyjasmonate with K(m) values of 50 and 10 microm, respectively. Furthermore, 12-hydroxyjasmonate and its sulfonated derivative are shown to be naturally occurring in A. thaliana. The exogenous application of methyljasmonate to A. thaliana plants led to increased levels of both metabolites, whereas treatment with 12-hydroxyjasmonate led to increased level of 12-hydroxyjasmonate sulfate without affecting the endogenous level of jasmonic acid. AtST2a expression was found to be induced following treatment with methyljasmonate and 12-hydroxyjasmonate. In contrast, the expression of the methyljasmonate-responsive gene Thi2.1, a marker gene in plant defense responses, is not induced upon treatment with 12-hydroxyjasmonate indicating the existence of independent signaling pathways responding to jasmonic acid and 12 hydroxyjasmonic acid. Taken together, the results suggest that the hydroxylation and sulfonation reactions might be components of a pathway that inactivates excess jasmonic acid in plants. Alternatively, the function of AtST2a might be to control the biological activity of 12-hydroxyjasmonic acid. PMID- 12637545 TI - Chromium (VI) activates ataxia telangiectasia mutated (ATM) protein. Requirement of ATM for both apoptosis and recovery from terminal growth arrest. AB - The ataxia telangiectasia mutated (ATM) protein plays a central role in early stages of DNA double strand break (DSB) detection and controls cellular responses to this damage. Although hypersensitive to ionizing radiation-induced clonogenic lethality, ataxia telangiectasia cells are paradoxically deficient in their ability to undergo ionizing radiation-induced apoptosis. This contradiction illustrates the complexity of the central role of ATM in DNA damage response and the need for further understanding. Certain hexavalent chromium (Cr(VI)) compounds are implicated as occupational respiratory carcinogens at doses that are both genotoxic and cytotoxic. Cr(VI) induces a broad spectrum of DNA damage, but Cr(VI)-induced DSBs have not been reported. Here, we examined the role of ATM in the cellular response to Cr(VI) and found that Cr(VI) activates ATM. We also show that physiological targets of ATM, p53 Ser-15 and Chk2 Thr-68, were phosphorylated by Cr(VI) exposure in an ATM-dependent fashion. We found that ATM /- cells were markedly resistant to Cr(VI)-induced apoptosis but considerably more sensitive to Cr(VI)-induced clonogenic lethality than wild type cells, indicating that resistance to Cr(VI)-induced apoptosis did not confer a selective survival advantage. However, analysis of long term growth arrest revealed a striking difference: ATM-/- cells were markedly less able to recover from Cr(VI) induced growth arrest. This indicates that terminal growth arrest is the fate of these apoptosis-resistant cells. In summary, ATM is involved in cellular response to a complex genotoxin that may not directly induce DSBs. Our data suggest that ATM is a major signal initiator for genotoxin-induced apoptosis but, paradoxically, also contributes to maintenance of cell survival by facilitating recovery/escape from terminal growth arrest. The results also strongly suggest that terminal growth arrest is not merely an extended or even irreversible form of checkpoint arrest, but instead an independent and unique cell fate pathway. PMID- 12637546 TI - Serp-1, a viral anti-inflammatory serpin, regulates cellular serine proteinase and serpin responses to vascular injury. AB - Complex DNA viruses have tapped into cellular serpin responses that act as key regulatory steps in coagulation and inflammatory cascades. Serp-1 is one such viral serpin that effectively protects virus-infected tissues from host inflammatory responses. When given as purified protein, Serp-1 markedly inhibits vascular monocyte invasion and plaque growth in animal models. We have investigated mechanisms of viral serpin inhibition of vascular inflammatory responses. In vascular injury models, Serp-1 altered early cellular plasminogen activator (tissue plasminogen activator), inhibitor (PAI-1), and receptor (urokinase-type plasminogen activator) expression (p < 0.01). Serp-1, but not a reactive center loop mutant, up-regulated PAI-1 serpin expression in human endothelial cells. Treatment of endothelial cells with antibody to urokinase-type plasminogen activator and vitronectin blocked Serp-1-induced changes. Significantly, Serp-1 blocked intimal hyperplasia (p < 0.0001) after aortic allograft transplant (p < 0.0001) in PAI-1-deficient mice. Serp-1 also blocked plaque growth after aortic isograft transplant and after wire-induced injury (p < 0.05) in PAI-1-deficient mice indicating that increase in PAI-1 expression is not required for Serp-1 to block vasculopathy development. Serp-1 did not inhibit plaque growth in uPAR-deficient mice after aortic allograft transplant. We conclude that the poxviral serpin, Serp-1, attenuates vascular inflammatory responses to injury through a pathway mediated by native uPA receptors and vitronectin. PMID- 12637547 TI - Ets-2 and components of mammalian SWI/SNF form a repressor complex that negatively regulates the BRCA1 promoter. AB - Ets-2 is a transcriptional activator that can be modulated by ras-dependent phosphorylation. Evidence is presented indicating that ets-2 can also act as a transcriptional repressor. In the breast cancer cell line MCF-7, exogenous ets-2 repressed the activity of a BRCA1 promoter-luciferase reporter dependent on a conserved ets-2-binding site in this promoter. Conditional overproduction of ets 2 in MCF-7 cells resulted in repression of endogenous BRCA1 mRNA expression. To address the mechanism by which ets-2 could act as a repressor, a biochemical approach was used to identify proteins that interacted with the ets-2 pointed domain. From this analysis, components of the mammalian SWI/SNF chromatin remodeling complex were found to interact with ets-2. Brg-1, the ATP-hydrolyzing component of the SWI/SNF complex, along with the BAF57/p50 and Ini1 subunits could be co-immunoprecipitated from cells with ets-2. The pointed domain of ets-2 directly interacted in vitro with the C-terminal region of Brg-1 in a phosphorylation-dependent manner. The combination of Brg-1 and ets-2 could repress the BRCA1 promoter reporter in transfection assays. These results support a role for ets-2 as a repressor and indicate that components of the mammalian SNF/SWI complex are required as co-repressors. PMID- 12637549 TI - In vitro regulation of budding yeast Bfa1/Bub2 GAP activity by Cdc5. AB - The Cdc5 protein of budding yeast is a polo-like kinase that has multiple roles in mitosis including control of the mitotic exit network (MEN). MEN activity brings about loss of mitotic kinase activity so that the mitotic spindle is disassembled and cytokinesis can proceed. Activity of the MEN is regulated by a small GTPase, Tem1, which in turn is controlled by a two-component GTPase activating protein (GAP) formed by Bfa1 and Bub2. Bfa1 has been identified as a regulatory target of Cdc5 but there are conflicting deductions from indirect in vivo assays as to whether phosphorylation inhibits or stimulates Bfa1 activity. To resolve this question, we have used direct in vitro assays to observe the effects of phosphorylation on Bfa1 activity. We show that when Bfa1 is phosphorylated by Cdc5, its GAP activity with Bub2 is inhibited although its ability to interact with Tem1 is unaffected. Thus, in vivo inactivation of Bfa1 Bub2 by Cdc5 would have a positive regulatory effect by increasing levels of Tem1 GTP so stimulating exit from mitosis. PMID- 12637548 TI - Prion infection impairs copper binding of cultured cells. AB - The molecular mechanism of neurodegeneration in transmissible spongiform encephalopathies (TSEs) remains unclear. Using radioactive copper ((64)Cu) at physiological concentration, we showed that prion infected cells display a marked reduction in copper binding. The level of full-length prion protein known to bind the metal ion was not modified in infected cells, but a fraction of this protein was not releasable from the membrane by phosphatidylinositol-specific phospholipase C. Our results suggest that prion infection modulates copper content at a cellular level and that modification of copper homeostasis plays a determinant role in the neuropathology of TSE. PMID- 12637550 TI - Phosphorylation of Tyr-176 of the yeast MAPK Hog1/p38 is not vital for Hog1 biological activity. AB - Mitogen-activated protein kinases are crucial components in the life of eukaryotic cells. The current dogma for MAPK activation is that dual phosphorylation of neighboring Thr and Tyr residues at the phosphorylation lip is an absolute requirement for their catalytic and biological activity. In this study we addressed the role of Tyr and Thr phosphorylation in the yeast MAPK Hog1/p38. Taking advantage of the recently isolated hyperactive mutants, whose intrinsic basal activity is independent of upstream regulation, we demonstrate that Tyr-176 is not required for basal catalytic and biological activity but is essential for the salt-induced amplification of Hog1 catalysis. We show that intact Thr-174 is absolutely essential for biology and catalysis of the mutants but is mainly required for structural reasons and not as a phosphoacceptor. The roles of Thr-174 and Tyr-176 in wild type Hog1 molecules were also tested. Unexpectedly we found that Hog1(Y176F) is biologically active, capable of induction of Hog1 target genes and of rescuing hog1Delta cells from osmotic stress. Hog1(Y176F) was not able, however, to mediate growth arrest induced by constitutively active MAPK kinase/Pbs2. We propose that Thr-174 is essential for stabilizing the basal active conformation, whereas Tyr-176 is not. Tyr-176 serves as a regulatory element required for stimuli-induced amplification of kinase activity. PMID- 12637551 TI - COX2 activity promotes organic osmolyte accumulation and adaptation of renal medullary interstitial cells to hypertonic stress. AB - The mechanism by which COX2 inhibition decreases renal cell survival is poorly understood. In the present study we examined the effect of COX2 activity on organic osmolyte accumulation in renal medulla and in cultured mouse renal medullary interstitial cells (MMICs) and its role in facilitating cell survival. Hypertonicity increased accumulation of the organic osmolytes inositol, sorbitol, and betaine in cultured mouse medullary interstitial cells. Pretreatment of MMICs with a COX2-specific inhibitor (SC58236, 10 micromol/liter) dramatically reduced osmolyte accumulation (by 79 +/- 9, 57 +/- 12, and 96 +/- 10% for inositol, sorbitol, and betaine respectively, p < 0.05). Similarly, 24 h of dehydration increased inner medullary inositol, sorbitol, and betaine concentrations in vivo by 85 +/- 10, 197 +/- 28, and 190 +/- 24 pmol/microg of protein, respectively, but this increase was also blunted (by 100 +/- 5, 66 +/- 15, and 81 +/- 9% for inositol, sorbitol, and betaine, respectively, p < 0.05) by pretreatment with an oral COX2 inhibitor. Dehydrated COX2-/- mice also exhibited an impressive defect in sorbitol accumulation (88 +/- 9% less than wild type, p < 0.05) after dehydration. COX2 inhibition (COX2 inhibitor-treated or COX2-/- MMICs) dramatically reduced the expression of organic osmolyte uptake mechanisms including betaine (BGT1) and sodium-myo-inositol transporter and aldose reductase mRNA expression under hypertonic conditions. Importantly, preincubation of COX2 inhibitor-treated MMICs with organic osmolytes restored their ability to survive hypertonic stress. In conclusion, osmolyte accumulation in the kidney inner medulla is dependent on COX2 activity, and providing exogenous osmolytes reverses COX2-induced cell death. These findings may have implications for the pathogenesis of analgesic nephropathy. PMID- 12637552 TI - Bacillus subtilis ResA is a thiol-disulfide oxidoreductase involved in cytochrome c synthesis. AB - Covalent attachment of heme to apocytochromes c in bacteria occurs on the outside of the cytoplasmic membrane and requires two reduced cysteinyls at the heme binding site. A constructed ResA-deficient Bacillus subtilis strain was found to lack c-type cytochromes. Cytochrome c synthesis was restored in the mutant by: (i) in trans expression of resA; (ii) deficiency in BdbD, a thiol-disulfide oxidoreductase that catalyzes formation of an intramolecular disulfide bond in apocytochrome c after transfer of the polypeptide across the cytoplasmic membrane; or (iii) by addition of the reductant dithiothreitol to the growth medium. In vivo studies of ResA showed that it is membrane-associated with its thioredoxin-like domain on the outside of the cytoplasmic membrane. Analysis of a soluble form of the protein revealed two redox reactive cysteine residues with a midpoint potential of about -340 mV at pH 7. We conclude that ResA, probably together with another thiol-disulfide oxidoreductase, CcdA, is required for the reduction of the cysteinyls in the heme binding site of apocytochrome c. PMID- 12637553 TI - Poly(ADP-ribose) polymerase-1 (PARP-1) is required in murine cell lines for base excision repair of oxidative DNA damage in the absence of DNA polymerase beta. AB - Oxidative DNA base damage is mainly corrected by the base excision repair (BER) pathway, which can be divided into two subpathways depending on the length of the resynthetized patch, either one nucleotide for short patch BER or several nucleotides for long patch BER. The role of proteins in the course of BER processes has been investigated in vitro using purified enzymes and cell-free extracts. In this study, we have investigated the repair of 8-oxo-7,8 dihydroguanine (8-oxoG) in vivo using wild-type, polymerase beta(-/-) (Polbeta(-/ )), poly(ADP-ribose) polymerase-1(-/-) (PARP-1(-/-)), and Polbeta(-/-)PARP-1(-/-) 3T3 cell lines. We used non replicating plasmids containing a 8-oxoG:C base pair to study the repair of the lesion located in a transcribed sequence (TS) or in a non-transcribed sequence (NTS). The results show that 8-oxoG repair in TS is not significantly impaired in cells deficient in Polbeta or PARP-1 or both. Whereas 8 oxoG repair in NTS is normal in Polbeta-null cells, it is delayed in PARP-1-null cells and greatly impaired in cells deficient in both Polbeta and PARP-1. The removal of 8-oxoG and presumably the cleavage at the resulting apurinic/apyrimidinic site are not affected in the PARP-1(-/-)Polbeta(-/-) cell lines. However, 8-oxoG repair is incomplete, yielding plasmid molecules with a nick at the site of the lesion. Therefore, PARP-1(-/-)Polbeta(-/-) cell lines cannot perform 5'-dRP removal and/or DNA repair synthesis. Furthermore, the poly(ADP-ribosyl)ation activity of PARP-1 is essential for 8-oxoG repair in a Polbeta(-/-) context, because expression of the catalytically inactive PARP-1 (E988K) mutant does not restore 8-oxoG repair, whereas an wild type PARP-1 does. PMID- 12637554 TI - Structural changes in RepA, a plasmid replication initiator, upon binding to origin DNA. AB - RepA protein is the DNA replication initiator of the Pseudomonas plasmid pPS10. RepA dimers bind to an inversely repeated operator sequence in repA promoter, thus repressing its own synthesis, whereas monomers bind to four directly repeated sequences (iterons) to initiate DNA replication. We had proposed previously that RepA is composed of two winged-helix (WH) domains, a structural unit also present in eukaryotic and archaeal initiators. To bind to the whole iteron sequence through both domains, RepA should couple monomerization to a conformational change in the N-terminal WH, which includes a leucine zipper-like sequence motif. We show for the first time that, by itself, binding to iteron DNA in vitro dissociates RepA dimers into monomers and alters RepA conformation, suggesting an allosteric effect. Furthermore, we also show that similar changes in RepA are promoted by mutations that substitute two Leu residues of the putative leucine zipper by Ala, destabilizing the hydrophobic core of the first WH. We propose that this mutant (RepA-2L2A) resembles a transient folding intermediate in the pathway leading to active monomers. These findings, together with the known activation of other Rep-type proteins by chaperones, are relevant to understand the molecular basis of plasmid DNA replication initiation. PMID- 12637556 TI - The RNA binding domains of the nuclear poly(A)-binding protein. AB - The nuclear poly(A)-binding protein (PABPN1) is involved in the synthesis of the mRNA poly(A) tails in most eukaryotes. We report that the protein contains two RNA binding domains, a ribonucleoprotein-type RNA binding domain (RNP domain) located approximately in the middle of the protein sequence and an arginine-rich C-terminal domain. The C-terminal domain also promotes self-association of PABPN1 and moderately cooperative binding to RNA. Whereas the isolated RNP domain binds specifically to poly(A), the isolated C-terminal domain binds non-specifically to RNA and other polyanions. Despite this nonspecific RNA binding by the C-terminal domain, selection experiments show that adenosine residues throughout the entire minimal binding site of approximately 11 nucleotides are recognized specifically. UV-induced cross-links with oligo(A) carrying photoactivatable nucleotides at different positions all map to the RNP domain, suggesting that most or all of the base-specific contacts are made by the RNP domain, whereas the C-terminal domain may contribute nonspecific contacts, conceivably to the same nucleotides. Asymmetric dimethylation of 13 arginine residues in the C-terminal domain has no detectable influence on the interaction of the protein with RNA. The N-terminal domain of PABPN1 is not required for RNA binding but is essential for the stimulation of poly(A) polymerase. PMID- 12637555 TI - Protective role of hydroxysteroid sulfotransferase in lithocholic acid-induced liver toxicity. AB - Supplement of 1% lithocholic acid (LCA) in the diet for 5-9 days resulted in elevated levels of the marker for liver damage aspartate aminotransferase and alkaline phosphatase activities in both farnesoid X receptor (FXR)-null and wild type female mice. The levels were clearly higher in wild-type mice than in FXR null mice, despite the diminished expression of a bile salt export pump in the latter. Consistent with liver toxicity marker activities, serum and liver levels of bile acids, particularly LCA and taurolithocholic acid, were clearly higher in wild-type mice than in FXR-null mice after 1% LCA supplement. Marked increases in hepatic sulfating activity for LCA (5.5-fold) and hydroxysteroid sulfotransferase (St) 2a (5.8-fold) were detected in liver of FXR-null mice. A 7.4-fold higher 3alpha-sulfated bile acid concentration was observed in bile of FXR-null mice fed an LCA diet compared with that of wild-type mice. Liver St2a content was inversely correlated with levels of alkaline phosphatase. In contrast, microsomal LCA 6beta-hydroxylation was not increased and was in fact lower in FXR-null mice compared in wild-type mice. Clear decreases in mRNA encoding sodium taurocholate cotransporting polypeptide, organic anion transporting polypeptide 1, and liver specific organic anion transporter-1 function in bile acid import were detected in LCA-fed mice. These transporter levels are higher in FXR-null mice than wild type mice after 1% LCA supplement. No obvious changes were detected in the Mrp2, Mrp3, and Mrp4 mRNAs. These results indicate hydroxysteroid sulfotransferase mediated LCA sulfation as a major pathway for protection against LCA-induced liver damage. Furthermore, Northern blot analysis using FXR-null, pregnane X receptor-null, and FXR-pregnane X receptor double-null mice suggests a repressive role of these nuclear receptors on basal St2a expression. PMID- 12637557 TI - Distinct function of conserved amino acids in the fingers of Saccharomyces cerevisiae DNA polymerase alpha. AB - Structural differences between class A and B DNA polymerases suggest that the motif B region, a wall of the catalytic pocket, may have evolved differentially in the two polymerase families. This study examines the function of the motif B residues in Saccharomyces cerevisiae DNA polymerase alpha (pol alpha). Effects of the mutations were determined by biochemical analysis and genetic complementation of a yeast strain carrying a temperature-sensitive pol alpha mutant. Many conserved residues were viable with a variety of substitutions. Among them, mutations at Asn-948 or Tyr-951 conferred up to 8-fold higher colony formation frequency in a URA3 forward mutation assay, and 79-fold higher trp1 reversion frequency was observed for Y951P in yeast. Purified Y951P was as accurate as wild type in DNA synthesis but approximately 6-fold less processive and 22-fold less active in vitro. Therefore, Y951P may increase the frequency of mutant colony formation because of its low level of DNA polymerase activity in yeast. Mutations at Lys-944 or Gly-952 were not viable, which is consistent with the observation that mutants with substitutions at Gly-952 have strongly reduced catalytic activity in vitro. Gly-952 may provide a space for the nascent base pair and thus may play an essential function in S. cerevisiae DNA pol alpha. These results suggest that class B DNA polymerases have a unique structure in the catalytic pocket, which is distinct from the corresponding region in class A DNA polymerases. PMID- 12637558 TI - The Gly-952 residue of Saccharomyces cerevisiae DNA polymerase alpha is important in discriminating correct deoxyribonucleotides from incorrect ones. AB - Gly-952 is a conserved residue in Saccharomyces cerevisiae DNA polymerase alpha (pol alpha) that is strictly required for catalytic activity and for genetic complementation of a pol alpha-deficient yeast strain. This study analyzes the role of Gly-952 by characterizing the biochemical properties of Gly-952 mutants. Analysis of the nucleotide incorporation specificity of pol alpha G952A showed that this mutant incorporates nucleotides with extraordinarily low fidelity. In a steady-state kinetic assay to measure nucleotide misincorporation, pol alpha G952A incorporated incorrect nucleotides more efficiently than correct nucleotides opposite template C, G, and T. The fidelity of the G952A mutant polymerase was highest at template A, where the ratio of incorporation of dCMP to dTMP was as high as 0.37. Correct nucleotide insertion was 500- to 3500-fold lower for G952A than for wild type pol alpha, with up to 22-fold increase in pyrimidine misincorporation. The Km for G952A pol alpha bound to mismatched termini T:T, T:C, C:A, and A:C was 71- to 460-fold lower than to a matched terminus. Furthermore, pol alpha G952A preferentially incorporated pyrimidine instead of dAMP opposite an abasic site, cis-syn cyclobutane di-thymine, or (6-4) di-thymine photoproduct. These data demonstrate that Gly-952 is a critical residue for catalytic efficiency and error prevention in S. cerevisiae pol alpha. PMID- 12637559 TI - High density lipoprotein-induced signaling of the MAPK pathway involves scavenger receptor type BI-mediated activation of Ras. AB - High density lipoprotein (HDL) stimulates multiple signaling pathways. HDL induced activation of the mitogen-activated protein kinase (MAPK) pathway can be mediated by protein kinase C (PKC) and/or pertussis toxin-sensitive G-proteins. Although HDL-induced activation of MAPK involves Raf-1, Mek, and Erk1/2, the upstream contribution of p21(ras) (Ras) on the activation of Raf-1 and MAPK remains elusive. Here we examine the effect of HDL on Ras activity and demonstrate that HDL induces PKC-independent activation of Ras that is completely blocked by pertussis toxin, thus implicating heterotrimeric G-proteins. In addition, the HDL-induced activation of Ras is inhibited by a neutralizing antibody against scavenger receptor type BI. We conclude that the binding of HDL to scavenger receptor type BI activates Ras in a PKC-independent manner with subsequent induction of the MAPK signaling cascade. PMID- 12637560 TI - Multiple interactions involving the amino-terminal domain of yeast mtRNA polymerase determine the efficiency of mitochondrial protein synthesis. AB - The amino-terminal domain (ATD) of Saccharomyces cerevisiae mitochondrial RNA polymerase has been shown to provide a functional link between transcription and post-transcriptional events during mitochondrial gene expression. This connection is mediated in large part by its interactions with the matrix protein Nam1p and, based on genetic phenotypes, the mitochondrial membrane protein Sls1p. These observations led us to propose previously that mtRNA polymerase, Nam1p, and Sls1p work together to coordinate transcription and translation of mtDNA-encoded gene products. Here we demonstrate by specific labeling of mitochondrial gene products in vivo that Nam1p and Sls1p indeed work together in a pathway that is required globally for efficient mitochondrial translation. Likewise, mutations in the ATD result in similar global reductions in mitochondrial translation efficiency and sensitivity to the mitochondrial translation inhibitor erythromycin. These data, coupled with the observation that the ATD is required to co-purify Sls1p in association with mtDNA nucleoids, suggest that efficient expression of mtDNA encoded genes in yeast involves a complex series of interactions that localize active transcription complexes to the inner membrane in order to coordinate translation with transcription. PMID- 12637561 TI - Increased sensitivity to endothelial nitric oxide (NO) contributes to arterial normotension in mice with vascular smooth muscle-selective deletion of the atrial natriuretic peptide (ANP) receptor. AB - Atrial natriuretic peptide (ANP) plays a key regulatory role in arterial blood pressure homeostasis. We recently generated mice with selective deletion of the ANP receptor, guanylyl cyclase-A (GC-A), in vascular smooth muscle (SMC GC-A knockout (KO) mice) and reported that resting arterial blood pressure was completely normal in spite of clear abolition of the direct vasodilating effects of ANP (Holtwick, R., Gotthardt, M., Skryabin, B., Steinmetz, M., Potthast, R., Zetsche, B., Hammer, R. E., Herz, J., and Kuhn M. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 7142-7147). The purpose of this study was to clarify mechanisms compensating for the missing vasodilator responses to ANP. In particular, we analyzed the effect of the endothelial, cGMP-mediated vasodilators C-type natriuretic peptide and nitric oxide (NO). In isolated arteries from SMC GC-A KO mice, the vasorelaxing sensitivity to sodium nitroprusside and the endothelium dependent vasodilator, acetylcholine, was significantly greater than in control mice. There was no difference in responses to C-type natriuretic peptide or to the activator of cGMP-dependent protein kinase I, 8-para-chlorophenylthio-cGMP. The aortic expression of soluble GC (sGC), but not of endothelial NO synthase or cGMP-dependent protein kinase I, was significantly increased in SMC GC-A KO mice. Chronic oral treatment with the NO synthase inhibitor N(w)-nitro-l-arginine methyl ester increased arterial blood pressure, the effect being significantly enhanced in SMC GC-A KO mice. We conclude that SMC GC-A KO mice exhibit a higher vasodilating sensitivity to NO. This can be attributed to an enhanced expression of sGC, whereas the expression and/or activity levels of downstream cGMP-effector pathways are not involved. Increased vasodilating responsiveness to endothelial NO contributes to compensate for the missing vasodilating effect of ANP in SMC GC A KO mice. PMID- 12637563 TI - The mitogen-activated protein kinase cascade promotes myoblast cell survival by stabilizing the cyclin-dependent kinase inhibitor, p21WAF1 protein. AB - During myogenesis, proliferating myoblasts withdraw from the cell cycle and are either eliminated by programmed cell death or differentiate into mature myotubes. Previous studies indicate that mitogen-activated protein kinase (MAPK) activity is significantly induced with the onset of terminal differentiation of C2 myoblasts. We have investigated the part played by the MAPK pathway in the differentiation of C2 myoblasts. Specific activation of MAPK by expression of an active Raf1-estrogen receptor chimera protein reduced significantly the number of myoblasts undergoing programmed cell death in the differentiation medium. Activation of Raf1 prevented the proteolytic activation of the proapoptotic caspase 9-protein during differentiation. The antiapoptotic function of Raf1 correlated with accumulation of the p21WAF1 protein resulting from its increased stability. Antisense expression of p21 was used to determine whether the p21WAF1 protein mediated the antiapoptotic activity of Raf1. Reduction of p21WAF1 protein in muscle cells abolished the antiapoptotic activity of the MAPK pathway. We conclude that MAPK contributes to muscle differentiation by preventing apoptotic cell death of differentiating myoblasts and that this activity is mediated by stabilization of the p21WAF1 protein. PMID- 12637562 TI - Immunopurification and characterization of rat adipocyte caveolae suggest their dissociation from insulin signaling. AB - Adipocytes play an important role in the insulin-dependent regulation of organismal fuel metabolism and express caveolae at levels as high or higher than any other cell type. Recently, a link between insulin signaling and caveolae has been suggested; nevertheless, adipocyte caveolae have been the subject of relatively few studies, and their contents have been minimally characterized. With the aid of a new monoclonal antibody, we developed a rapid procedure for the immunoisolation of caveolae derived from the plasma membrane of adipocytes, and we characterized their protein content. We find that immunopurified adipocyte caveolae have a relatively limited protein composition, and they lack the raft protein, flotillin, and insulin receptors. Immunogold labeling and electron microscopy of the adipocyte plasma membrane confirmed the lack of insulin receptors in caveolae. In addition to caveolins, the structural components of caveolae, their major protein constituents, are the semicarbazide-sensitive amine oxidase and the scavenger lipoprotein receptor CD36. The results are consistent with a role for caveolae in lipid flux in and of adipocytes. PMID- 12637564 TI - Maturation of the regulation of GLUT4 activity by p38 MAPK during L6 cell myogenesis. AB - Insulin stimulates glucose uptake in skeletal muscle cells and fat cells by promoting the rapid translocation of GLUT4 glucose transporters to the plasma membrane. Recent work from our laboratory supports the concept that insulin also stimulates the intrinsic activity of GLUT4 through a signaling pathway that includes p38 MAPK. Here we show that regulation of GLUT4 activity by insulin develops during maturation of skeletal muscle cells into myotubes in concert with the ability of insulin to stimulate p38 MAPK. In L6 myotubes expressing GLUT4 that carries an exofacial myc-epitope (L6-GLUT4myc), insulin-stimulated GLUT4myc translocation equals in magnitude the glucose uptake response. Inhibition of p38 MAPK with SB203580 reduces insulin-stimulated glucose uptake without affecting GLUT4myc translocation. In contrast, in myoblasts, the magnitude of insulin stimulated glucose uptake is significantly lower than that of GLUT4myc translocation and is insensitive to SB203580. Activation of p38 MAPK by insulin is considerably higher in myotubes than in myoblasts, as is the activation of upstream kinases MKK3/MKK6. In contrast, the activation of all three Akt isoforms and GLUT4 translocation are similar in myoblasts and myotubes. Furthermore, GLUT4myc translocation and phosphorylation of regulatory sites on Akt in L6 GLUT4myc myotubes are equally sensitive to insulin, whereas glucose uptake and phosphorylation of regulatory sites on p38 MAPK show lower sensitivity to the hormone. These observations draw additional parallels between Akt and GLUT4 translocation and between p38 MAPK and GLUT4 activation. Regulation of GLUT4 activity by insulin develops upon muscle cell differentiation and correlates with p38 MAPK activation by insulin. PMID- 12637565 TI - The Nucleo-cytoplasmic actin-binding protein CapG lacks a nuclear export sequence present in structurally related proteins. AB - Despite thorough structure-function analyses, it remains unclear how CapG, a ubiquitous F-actin barbed end capping protein that controls actin microfilament turnover in cells, is able to reside in the nucleus and cytoplasm, whereas structurally related actin-binding proteins are predominantly cytoplasmic. Here we report the molecular basis for the different subcellular localization of CapG, severin, and fragminP. Green fluorescent protein-tagged fragminP and severin accumulate in the nucleus upon treatment of transfected cells with the CRM1 inhibitor leptomycin B. We identified a nuclear export sequence in severin and fragminP, which is absent in CapG. Deletion of amino acids Met(1)-Leu(27) resulted in nuclear accumulation of severin and fragminP. Tagging this sequence to CapG triggered nuclear export, whereas mutation of single leucine residues (Leu(17), Leu(21), and Leu(27)) in the export sequence inhibited nuclear export. Based on these findings, a nuclear export signal was identified in myopodin, a muscle-specific actin-binding protein, and the Bloom syndrome protein, a RecQ like helicase. Deletion of the myopodin nuclear export sequence blocked invasion into collagen type I of C2C12 cells transiently overexpressing myopodin. Our findings explain regulated subcellular targeting of distinct classes of actin binding proteins. PMID- 12637566 TI - Caldesmon inhibits Arp2/3-mediated actin nucleation. AB - The Arp2/3 complex greatly accelerates actin polymerization, which is thought to play a major role in cell motility by inducing membrane protrusions including ruffling movements. Membrane ruffles contain a variety of actin-binding proteins, which would modulate Arp2/3-dependent actin polymerization. However, their exact roles in actin polymerization remain to be established. Because caldesmon is present in membrane ruffles, as well as in stress fibers, it may alter Arp2/3 mediated actin polymerization. We have found that caldesmon greatly retards Arp2/3-induced actin polymerization. Kinetic analyses have revealed that caldesmon inhibits the nucleation process, whereas it does not largely reduce elongation. Caldesmon is found to inhibit binding of Arp2/3 to F-actin, which apparently reduces the ability of F-actin as a secondary activator of Arp2/3 mediated nucleation. We also have found that the inhibition of the binding between actin and caldesmon either by Ca(2+)/calmodulin or by phosphorylation with cdc2 kinase reverses the inhibitory effect of caldesmon on Arp2/3-induced actin polymerization. Our results suggest that caldesmon may be a key protein that modulates membrane ruffling and that this may involve changes in caldesmon phosphorylation and/or intracellular calcium concentrations during signal transduction. PMID- 12637568 TI - Insulin-stimulated phosphorylation of a Rab GTPase-activating protein regulates GLUT4 translocation. AB - Insulin stimulates the rapid translocation of intracellular glucose transporters of the GLUT4 isotype to the plasma membrane in fat and muscle cells. The connections between known insulin signaling pathways and the protein machinery of this membrane-trafficking process have not been fully defined. Recently, we identified a 160-kDa protein in adipocytes, designated AS160, that is phosphorylated by the insulin-activated kinase Akt. This protein contains a GTPase-activating domain (GAP) for Rabs, which are small G proteins required for membrane trafficking. In the present study we have identified six sites of in vivo phosphorylation on AS160. These sites lie in the motif characteristic of Akt phosphorylation, and insulin treatment increased phosphorylation at five of the sites. Expression of AS160 with two or more of these sites mutated to alanine markedly inhibited insulin-stimulated GLUT4 translocation in 3T3-L1 adipocytes. Moreover, this inhibition did not occur when the GAP function in the phosphorylation site mutant was inactivated by a point mutation. These findings strongly indicate that insulin-stimulated phosphorylation of AS160 is required for GLUT4 translocation and that this phosphorylation signals translocation through inactivation of the Rab GAP function. PMID- 12637567 TI - Transcriptional regulation of heme oxygenase-1 gene expression by MAP kinases of the JNK and p38 pathways in primary cultures of rat hepatocytes. AB - Heme oxygenase-1 (HO-1) gene expression is induced by various oxidative stress stimuli including sodium arsenite. Since mitogen-activated protein kinases (MAPKs) are involved in stress signaling we investigated the role of arsenite and MAPKs for HO-1 gene regulation in primary rat hepatocytes. The Jun N-terminal kinase (JNK) inhibitor SP600125 decreased sodium arsenite-mediated induction of HO-1 mRNA expression. HO-1 protein and luciferase activity of reporter gene constructs with -754 bp of the HO-1 promoter were induced by overexpression of kinases of the JNK pathway and MKK3. By contrast, overexpression of Raf-1 and ERK2 did not affect expression whereas overexpression of p38alpha, beta, and delta decreased and p38gamma increased HO-1 expression. Electrophoretic mobility shift assays (EMSA) revealed that a CRE/AP-1 element (-668/-654) bound c-Jun, a target of the JNK pathway. Deletion or mutation of the CRE/AP-1 obliterated the JNK- and c-Jun-dependent up-regulation of luciferase activity. EMSA also showed that an E-box (-47/-42) was bound by a putative p38 target c-Max. Mutation of the E-box strongly reduced MKK3, p38 isoform-, and c-Max-dependent effects on luciferase activity. Thus, the HO-1 CRE/AP-1 element mediates HO-1 gene induction via activation of JNK/c-Jun whereas p38 isoforms act through a different mechanism via the E-box. PMID- 12637569 TI - Paired basic/Furin-like proprotein convertase cleavage of Pro-BMP-1 in the trans Golgi network. AB - Bone morphogenetic protein (BMP)-1 is a zinc-dependent metalloproteinase that cleaves a variety of extracellular matrix substrates, including type I procollagen. Little is known about the site of action of BMP-1, although the extracellular matrix seems likely to be it. BMP-1 is synthesized with an N terminal prodomain. The removal of the prodomain presumably activates the proteinase. In this study we show that the prodomain is cleaved in the trans Golgi network (TGN) and by furin-like/paired basic proprotein convertases. Inhibitors of furin resulted in the secretion of pro-BMP-1, which could not cleave procollagen. Recombinant furin cleaved the prodomain from pro-BMP-1. Site directed mutagenesis of the prodomain cleavage site (RSRR) to RSAA resulted in efficient secretion of pro-BMP-1. Therefore, prodomain cleavage was not required for secretion. Using peptide N-glycosidase and neuraminidase digestion to determine the post-translational status of pro-BMP-1 during its conversion to BMP 1, we showed that BMP-1 first appears in the TGN during sialylation of the molecule. Furthermore, immunofluorescence studies using an antibody to the nascent N terminus of BMP-1 showed localization to the TGN and plasma membrane. The observation that BMP-1 occurs inside the cell raises the possibility that BMP 1 might begin to cleave its substrates prior to secretion to the extracellular matrix. PMID- 12637570 TI - Membrane rafts play a crucial role in receptor activator of nuclear factor kappaB signaling and osteoclast function. AB - Membrane lipid rafts play a key role in immune cell activation by recruiting and excluding specific signaling components of immune cell surface receptors upon the receptor engagement. Despite this, the role of these microdomains in the regulation of osteoclasts as controlled by receptor activator of nuclear factor kappaB (RANK) has yet to be established. In this study, we demonstrate that the raft microdomain expression plays an essential role in osteoclast function and differentiation. Expression of raft component flotillin greatly increased during osteoclast differentiation, whereas engagement of RANK induced the translocation of tumor necrosis factor receptor-associated factor 6 to rafts where Src was constitutively resident. Disruption of rafts blocked TRAF6 translocation and Akt activation by RANK ligand in osteoclasts and further reduced the survival of osteoclasts. Actin ring formation and bone resorption by osteoclasts were also found to require the integrity of rafts. Our observations demonstrate for the first time that RANK-mediated signaling and osteoclast function are critically dependent on the expression and integrity of raft membrane microdomains. PMID- 12637571 TI - Regulation of vascular endothelial growth factor binding and activity by extracellular pH. AB - Angiogenesis, the growth of new blood vessels, is regulated by a number of factors, including hypoxia and vascular endothelial growth factor (VEGF). Although the effects of hypoxia have been studied intensely, less attention has been given to other extracellular parameters such as pH. Thus, the present study investigates the consequences of acidic pH on VEGF binding and activity in endothelial cell cultures. We found that the binding of VEGF165 and VEGF121 to endothelial cells increased as the extracellular pH was decreased from 7.5 to 5.5. Binding of VEGF165 and VEGF121 to endothelial extracellular matrix was also increased at acidic pH. These effects were, in part, a reflection of increased heparin binding, because VEGF165 and VEGF121 showed increased retention on heparin-Sepharose at pH 5.5 compared with pH 7.5. Consistent with these findings, soluble heparin competed for VEGF binding to endothelial cells under acidic conditions. However, at neutral pH (7.5) low concentrations of heparin (0.1-1.0 microg/ml) potentiated VEGF binding. Extracellular pH also regulated VEGF activation of the extracellular signal-regulated kinases 1 and 2 (Erk1/2). VEGF165 and VEGF121 activation of Erk1/2 at pH 7.5 peaked after 5 min, whereas at pH 6.5 the peak was shifted to 10 min. At pH 5.5, neither VEGF isoform was able to activate Erk1/2, suggesting that the increased VEGF bound to the cells at low pH was sequestered in a stored state. Therefore, extracellular pH might play an important role in regulating VEGF interactions with cells and the extracellular matrix, which can modulate VEGF activity. PMID- 12637572 TI - Polyclonal anti-PrP auto-antibodies induced with dimeric PrP interfere efficiently with PrPSc propagation in prion-infected cells. AB - Prion diseases are neurodegenerative infectious disorders for which no prophylactic regimens are known. In order to induce antibodies/auto-antibodies directed against surface-located PrP(c), we used a covalently linked dimer of mouse prion protein expressed recombinantly in Escherichia coli. Employing dimeric PrP as an immunogen we were able to effectively overcome autotolerance against murine PrP in PrP wild-type mice without inducing obvious side effects. Treatment of prion-infected mouse cells with polyclonal anti-PrP antibodies generated in rabbit or auto-antibodies produced in mice significantly inhibited endogenous PrP(Sc) synthesis. We show that polyclonal antibodies are binding to surface-located PrP(c), thereby interfering with prion biogenesis. This effect is much more pronounced in the presence of full IgG molecules, which, unlike Fab fragments, seem to induce a significant cross-linking of surface PrP. In addition, we found immune responses against different epitopes when comparing antibodies induced in rabbits and PrP wild-type mice. Only in the auto-antibody situation in mice an immune reaction against a region of PrP is found that was reported to be involved in the PrP(Sc) conversion process. Our data point to the possibility of developing means for an active immunoprophylaxis against prion diseases. PMID- 12637573 TI - Dominant-negative IkappaB facilitates apoptosis of osteoclasts by tumor necrosis factor-alpha. AB - Osteoclasts are the sole bone-resorbing cells. Heightened activity of these cells under pathological conditions leads to the development of bone loss diseases, such as osteolysis, osteoporosis, and rheumatoid arthritis. We have shown previously that tumor necrosis factor alpha-(TNF) strongly induces osteoclastogenesis of preosteoclasts and do so through activation of the transcription factor, NF-kappaB. Most importantly, recent studies have shown that NF-kappaB is required for the development of osteoclasts. This transcription factor has also been proven as an essential mediator of inflammatory diseases including those related to bone. In this regard, we have shown that various mutated forms of IkappaBalpha are potent inhibitors of osteoclastogenesis. In this study, we examined the direct effect of DN-IkappaB on mature and preosteoclast development in the presence of TNF. Our findings indicate that once committed to the osteoclastogenic pathway, preosteoclasts form giant and hyperactive osteoclasts in response to TNF. However, administration of DN-IkappaB to cultures prior to TNF exposure averts the osteoclastogenic effect of TNF into apoptosis. Screening potential mediators of DN-IkappaB and TNF-induced apoptosis shows that caspase 3, caspase 9, poly(ADP-ribose)polymerase, and Bax are activated, whereas levels of Bcl-XL, cIAP-1, and TRAF6 were reduced. Taken together, these findings suggest that under conditions of NF-kappaB inactivity levels of pro-survival factors are diminished, which in turn facilitates TNF induction of pro-apoptotic factors leading to apoptosis. PMID- 12637575 TI - Induction of adherent activity in mastocytoma P-815 cells by the cooperation of two prostaglandin E2 receptor subtypes, EP3 and EP4. AB - In this study, we investigated the role of PGE(2) in mouse mastocytoma P-815 cell adhesion to extracellular matrix proteins (ECMs) in vitro. We report that PGE(2) accelerated ProNectin F(TM) (a proteolytic fragment of fibronectin)-mediated adhesion, which was abolished by addition of the GRGDS peptide, an inhibitor of the RDG binding site of ProNectin F(TM). We show that the cAMP level and cAMP regulated protein kinase (PKA) activity are critical mediators of this PGE(2) effect, because the cell-permeable cAMP analogue 8-Br-cAMP accelerated P-815 cell adhesion to ProNectin F(TM) and the pharmacological inhibitor of PKA, H-89, blocked PGE(2)-mediated adhesion. Consistent with mRNA expression of the G(s) coupled EP4- and G(i)-coupled EP3-PGE receptor subtypes, P-815 cell adhesion was accelerated by treatment with a selective EP4 agonist, ONO-AE1-329, but not a selective EP1/EP3 agonist, sulprostone. However, simultaneous treatment with ONO AE1-329 and sulprostone resulted in augmentation of both the cAMP level and cell adhesion. The augmentation of EP3-mediated cAMP synthesis was dose-dependent, without affecting the half-maximal concentration for EP4-mediated G(s)-activity, which was inhibited by a G(i) inhibitor, pertussis toxin. In conclusion, these findings suggest that PGE(2) accelerates RGD-dependent adhesion via cooperative activation between EP3 and EP4 and contributes to the recruitment of mast cells to the ECM during inflammation. PMID- 12637574 TI - Egr-1 mediates transcriptional repression of COL2A1 promoter activity by interleukin-1beta. AB - Following induction and activation of the early growth response (Egr)-1 transcription factor in human chondrocytes, interleukin-1beta (IL-1beta) suppresses the expression of the type II collagen gene (COL2A1), associated with induction of Egr-1 binding activity in nuclear extracts. The COL2A1 proximal promoter contains overlapping binding sites for Egr-1 and Sp1 family members at 119/-112 bp and -81/-74 bp. Mutations that block binding of Sp1 and Sp3 to either site markedly reduce constitutive expression of the core promoter. IL-1beta induced Egr-1 binds strongly to the -119/-112 bp site, and mutations that block Egr-1 binding prevent inhibition by IL-1beta. Cotransfection with pCMV-Egr1 potentiates the inhibition of COL2A1 promoter activity by IL-1beta, whereas overexpression of dominant-negative Egr-1 mutant, Wilm's tumor-1 (WT1)/Egr1, Sp1, or Sp3 reverses the inhibition by IL-1beta. Cotransfection of pGL2-COL2/Gal4, in which we substituted the critical residue for Egr-1 binding with a Gal4 binding domain and a pCMV-Gal4-Egr1 chimera permits an inhibitory response to IL-1beta that is reversed by overexpression of Gal4-CBP. Our results indicate that IL 1beta-induced activation of Egr-1 binding is required for inhibition of COL2A1 proximal promoter activity and suggest that Egr-1 acts as a repressor of a constitutively expressed collagen gene by preventing interactions between Sp1 and the general transcriptional machinery. PMID- 12637576 TI - Levuglandinyl adducts of proteins are formed via a prostaglandin H2 synthase dependent pathway after platelet activation. AB - The product of oxygenation of arachidonic acid by the prostaglandin H synthases (PGHS), prostaglandin H(2) (PGH(2)), undergoes rearrangement to the highly reactive gamma-ketoaldehydes, levuglandin (LG) E(2), and LGD(2). We have demonstrated previously that LGE(2) reacts with the epsilon-amine of lysine to form both the levuglandinyl-lysine Schiff base and the pyrrole-derived levuglandinyl-lysine lactam adducts. We also have reported that these levuglandinyl-lysine adducts are formed on purified PGHSs following the oxygenation of arachidonic acid. We now present evidence that the levuglandinyl lysine lactam adduct is formed in human platelets upon activation with exogenous arachidonic acid or thrombin. After proteolytic digestion of the platelet proteins, and isolation of the adducted amino acid residues, this adduct was identified by liquid chromatography-tandem mass spectrometry. We also demonstrate that formation of these adducts is inhibited by indomethacin, a PGHS inhibitor, and is enhanced by an inhibitor of thromboxane synthase. These data establish that levuglandinyl-lysine adducts are formed via a PGHS-dependent pathway in whole cells, even in the presence of an enzyme that metabolizes PGH(2). They also demonstrate that a physiological stimulus is sufficient to lead to the lipid modification of proteins through the levuglandin pathway in human platelets. PMID- 12637577 TI - Defining the involvement of p38alpha MAPK in the production of anti- and proinflammatory cytokines using an SB 203580-resistant form of the kinase. AB - Despite its lack of specificity, the inhibitor SB 203580 has been widely used to implicate p38 mitogen-activated protein kinase (MAPK) in the synthesis of many cytokines. Here we show unequivocally that the production of interleukin (IL) 1beta, IL-6, IL-10, and tumor necrosis factor alpha (TNFalpha) requires p38 MAPK activity by demonstrating that the inhibitory effects of SB 203580 were reversed by expression of an SB 203580-resistant form of p38alpha (SBR-p38alpha) that fails to bind to SB 203580. This strategy established the requirement for p38 activity for the lipopolysaccharide-stimulated production of IL-10, IL-1beta, and IL-6 by the monocytic cell WEHI 274 and the production of IL-6 and TNFalpha stimulated by ligation of the Fc-gamma receptor of the mast cell MC/9. Expression of SBR-p38alpha in primary macrophages abrogated the ability of SB 203580 to inhibit the lipopolysaccharide-stimulated production of TNFalpha but not of IL 10. Expression of SBR-p38alpha in primary T lymphocytes abrogated the ability of SB 203580 to inhibit the production of interferon-gamma induced by co-ligation of CD3 and CD28 but not the production of interferon-gamma or IL-10 induced by IL 12. These results suggest that the levels of p38 MAPK activity required for maximal cytokine production vary with different cytokines and stimuli. PMID- 12637578 TI - Signaling role of intracellular iron in NF-kappaB activation. AB - Iron chelators inhibit endotoxin-induced NF-kappaB activation in hepatic macrophages (HMs), suggesting a role for the intracellular chelatable pool of iron in NF-kappaB activation. The present study tested this hypothesis. Analysis of Fe(59)-loaded HMs stimulated with lipopolysaccharide (LPS), revealed a previously unreported, transient rise in intracellular low molecular weight (LMW).Fe(59) complex ([LMW.Fe](i)) at /=15 min) and NF-kappaB (>/=30 min) activation. Iron chelators (1,2-dimethyl-3 hydroxypyridin-4-one and N,N'-bis-2-hydroxybenzylethylenediamine-N,N'-diacetic acid) abrogated the [LMW.Fe](i) response and IKK and NF-kappaB activation. The [LMW.Fe](i) response was also observed in tumor necrosis factor alpha (TNFalpha) stimulated HMs and RAW264.7 cells treated with LPS and interferon-gamma but not in primary rat hepatocytes or myofibroblastic cells exposed to LPS or TNFalpha. Both [LMW.Fe](i) response and IKK activation in LPS-stimulated HMs were inhibited by diphenylene iodonium (nonspecific inhibitor for flavin-containing oxidases), l N(6)-(1-iminoethyl)lysine (selective iNOS inhibitor), and adenoviral-mediated expression of a dominant negative mutant of Rac1 or Cu,Zn-superoxide dismutase, suggesting the role of (.)NO and O(2)() in mediating the iron signaling. In fact, this inhibition was recapitulated by a cell-permeable scavenger of ONOO(-), 5,10,15,20-tetrakis (4-sulfonatophenyl)porphyrinato iron (III) chloride. Conversely, ONOO(-) alone induced both [LMW.Fe](i) response and IKK activation. Finally, direct addition of ferrous iron to cultured HMs activated IKK and NF kappaB. These results support a novel signaling role for [LMW.Fe](i) in IKK activation, which appears to be induced by ONOO(-) and selectively operative in macrophages. PMID- 12637579 TI - A role for native lipids in the stabilization and two-dimensional crystallization of the Escherichia coli NADH-ubiquinone oxidoreductase (complex I). AB - NADH-ubiquinone oxidoreductase (complex I or NDH-1) was purified from the BL21 strain of Escherichia coli using an improved procedure. The complex was effectively stabilized by addition of divalent cations and lipids, making the preparation suitable for structural studies. The ubiquinone reductase activity of the enzyme was fully restored by addition of native E. coli lipids. Two different two-dimensional crystal forms, with p2 and p3 symmetry, were obtained using lipids containing native E. coli extracts. Analysis of the crystals showed that they are formed by fully intact complex I in an L-shaped conformation. Activity assays and single particle analysis indicated that complex I maintains this structure in detergent solution and does not adopt a different conformation in the active state. Thus, we provide the first experimental evidence that complex I from E. coli has an L-shape in a lipid bilayer and confirm that this is also the case for the active enzyme in solution. This suggests strongly that bacterial complex I exists in an L-shaped conformation in vivo. Our results also indicate that native lipids play an important role in the activation, stabilization and, as a consequence, crystallization of purified complex I from E. coli. PMID- 12637580 TI - Reb1p-dependent DNA bending effects nucleosome positioning and constitutive transcription at the yeast profilin promoter. AB - The molecular basis of constitutive gene activation is largely unknown. The yeast profilin gene (PFY1), encoding a housekeeping component of the actin cytoskeleton, is constitutively transcribed at a moderate level. The PFY1 promoter dispenses with classical transactivators and a consensus TATA box; however, it contains a canonic site for the abundant multifunctional nuclear factor rDNA enhancer-binding protein (Reb1p) combined with a dA.dT element. Reb1p binds specifically in vitro. Mutation of this site reduces PFY1 expression to about 35%. A nucleosome-free gap of about 190 bp is centered at the genomic Reb1p binding site in vivo and spans the presumptive core promoter and transcriptional initiation sites. Nucleosomes at the border of the gap are positioned. Mutation of the Reb1p motif in the genomic PFY1 promoter abolishes nucleosome positioning, fills the gap with a non-positioned nucleosome, and reduces transcription by a factor of 3. From permutation studies we conclude that Reb1p induces a strong bend into the DNA. Phasing analyses indicate that it is directed toward the major groove. The data suggest that Reb1p plays an architectural role on DNA and that Reb1p-dependent DNA bending leads to a DNA conformation that is incompatible with packaging into nucleosomes and concomitantly facilitates constitutive transcription. In the absence of other transcription activators, Reb1p excludes nucleosomes and moderately stimulates transcription by distorting DNA. PMID- 12637581 TI - Sulindac sulfide is a noncompetitive gamma-secretase inhibitor that preferentially reduces Abeta 42 generation. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) have been known to reduce risk for Alzheimer's disease. In addition to the anti-inflammatory effects of NSAIDs to block cylooxygenase, it has been shown recently that a subset of NSAIDs selectively inhibits the secretion of highly amyloidogenic Abeta42 from cultured cells, although the molecular target(s) of NSAIDs in reducing the activity of gamma-secretase for Abeta42 generation (gamma(42)-secretase) still remain unknown. Here we show that sulindac sulfide (SSide) directly acts on gamma secretase and preferentially inhibits the gamma(42)-secretase activity derived from the 3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-1-propanesulfonate solubilized membrane fractions of HeLa cells, in an in vitro gamma-secretase assay using recombinant amyloid beta precursor protein C100 as a substrate. SSide also inhibits activities for the generation of Abeta40 as well as for Notch intracellular domain at higher concentrations. Notably, SSide displayed linear noncompetitive inhibition profiles for gamma(42)-secretase in vitro. Our data suggest that SSide is a direct inhibitor of gamma-secretase that preferentially affects the gamma(42)-secretase activity. PMID- 12637582 TI - Crystal structure of Mycobacterium tuberculosis diaminopimelate decarboxylase, an essential enzyme in bacterial lysine biosynthesis. AB - The Mycobacterium tuberculosis lysA gene encodes the enzyme meso-diaminopimelate decarboxylase (DAPDC), a pyridoxal-5'-phosphate (PLP)-dependent enzyme. The enzyme catalyzes the final step in the lysine biosynthetic pathway converting meso-diaminopimelic acid (DAP) to l-lysine. The lysA gene of M. tuberculosis H37Rv has been established as essential for bacterial survival in immunocompromised mice, demonstrating that de novo biosynthesis of lysine is essential for in vivo viability. Drugs targeted against DAPDC could be efficient anti-tuberculosis drugs, and the three-dimensional structure of DAPDC from M. tuberculosis complexed with reaction product lysine and the ternary complex with PLP and lysine in the active site has been determined. The first structure of a DAPDC confirms its classification as a fold type III PLP-dependent enzyme. The structure shows a stable 2-fold dimer in head-to-tail arrangement of a triose phosphate isomerase (TIM) barrel-like alpha/beta domain and a C-terminal beta sheet domain, similar to the ornithine decarboxylase (ODC) fold family. PLP is covalently bound via an internal aldimine, and residues from both domains and both subunits contribute to the binding pocket. Comparison of the structure with eukaryotic ODCs, in particular with a di-fluoromethyl ornithine (DMFO)-bound ODC from Trypanosoma bruceii, indicates that corresponding DAP-analogues might be potential inhibitors for mycobacterial DAPDCs. PMID- 12637583 TI - Secretory IgA N- and O-glycans provide a link between the innate and adaptive immune systems. AB - Secretory IgA (SIgA) is a multi-polypeptide complex consisting of a secretory component (SC) covalently attached to dimeric IgA containing one joining (J) chain. We present the analysis of both the N- and O-glycans on the individual peptides from this complex. Based on these data, we have constructed a molecular model of SIgA1 with all its glycans, in which the Fab arms form a T shape and the SC is wrapped around the heavy chains. The O-glycan regions on the heavy (H) chains and the SC N-glycans have adhesin-binding glycan epitopes including galactose-linked beta1-4 and beta1-3 to GlcNAc, fucose-linked alpha1-3 and alpha1 4 to GlcNAc and alpha1-2 to galactose, and alpha2-3 and alpha2-6-linked sialic acids. These glycan epitopes provide SIgA with further bacteria-binding sites in addition to the four Fab-binding sites, thus enabling SIgA to participate in both innate and adaptive immunity. We also show that the N-glycans on the H chains of both SIgA1 and SIgA2 present terminal GlcNAc and mannose residues that are normally masked by SC, but that can be unmasked and recognized by mannose-binding lectin, by disrupting the SC-H chain noncovalent interactions. PMID- 12637584 TI - CBP recruitment and histone acetylation in differential gene induction by glucocorticoids and progestins. AB - We have analyzed histone acetylation at the steroid-responsive mouse mammary tumor virus (MMTV) promoter in five separate cell lines that express functional glucocorticoid and/or progesterone receptors. Chromatin immunoprecipitation assays reveal that glucocorticoid and progesterone receptors bind the MMTV promoter after hormone addition but that receptor binding is not associated with an increase in acetylation of histone H3 or H4. We have, however, found one exception to this rule. Previously we described a cell line [T47D(C&L)] that displayed a remarkable differential induction of MMTV by glucocorticoids and progestins. At one chromosomal locus (MMTV-luciferase), MMTV is preferentially induced by glucocorticoids, whereas at another locus within the same cell (MMTV CAT), MMTV is activated by both glucocorticoids and progestins. Here we show that the glucocorticoid-mediated induction of MMTV-luciferase is accompanied by increased recruitment of CBP to the promoter and increased histone H3 and H4 acetylation, whereas the hormonal induction of MMTV-CAT in the same cell exhibits a more modest CBP recruitment without any increase in histone acetylation. These studies suggest that increased histone acetylation may serve a potentiating function for MMTV promoter activation at certain loci. However, increased histone acetylation is not requisite for steroid-mediated induction of transcription at all genes. PMID- 12637585 TI - Cellular context of coregulator and adaptor proteins regulates human adenovirus 5 early region 1A-dependent gene activation by the thyroid hormone receptor. AB - In mammalian cells, the human adenovirus type 5 early region 1A (E1A) oncoprotein functions as a thyroid hormone (TH)-dependent activator of the thyroid hormone receptor (TR). Interestingly, in the cellular context of the yeast Saccharomyces cerevisiae, E1A acts as a TR-specific constitutive coactivator that is down regulated by TH. TH reduces the interaction of E1A with the TR in yeast but not HeLa cells. The N-terminal 82 amino acids of E1A are sufficient for coactivation in yeast and residues 4-29 are essential. In yeast, expression of the nuclear receptor corepressor (N-CoR) could down-regulate constitutive transcriptional activation of the TR by E1A, whereas expression of the glucocorticoid receptor interacting protein 1 (GRIP-1) coactivator reconstituted the E1A-induced pattern of enhanced TH-dependent gene activation by TR observed in mammalian cells. We further show that the mating type switching gene (SWI)/sucrose nonfermenting (SNF) gene chromatin remodeling complex is required for both TH/GRIP-1- and E1A dependent coactivator function, whereas the general control nonrepressed protein (GCN5)/alteration/deficiency in activation protein (ADA2) components of the SPT, ADA, GCN5, acetylation (SAGA) transcriptional adaptor complex are required for TH/GRIP-1, but not E1A-dependent activation of the TR. Taken together, these studies demonstrate that the novel TR-specific coactivator function of E1A in yeast depends on the SWI/SNF chromatin remodeling complex and can be further influenced by changes in the cellular complement of transcriptional coregulatory proteins. PMID- 12637586 TI - Activation of signal transducer and activator of transcription 3 by oncogenic RET/PTC (rearranged in transformation/papillary thyroid carcinoma) tyrosine kinase: roles in specific gene regulation and cellular transformation. AB - Thyroid papillary carcinomas are characterized by RET/PTC (rearranged in transformation/papillary thyroid carcinoma) rearrangements that result in fusion of the tyrosine kinase domain of the RET receptor to the N-terminal sequences encoded by heterologous genes. This thyroid-specific rearrangement causes aberrant expression of RET/PTC and results in constitutive ligand-independent activation of RET kinase. However, it is unclear how RET/PTC activates the specific signaling pathways for cellular transformation. In this study, we show that RET/PTC associates with signal transducer and activator of transcription 3 (STAT3) and activates it by the specific phosphorylation of the tyrosine 705 residue. Activation of STAT3 requires the intrinsic kinase activity of RET/PTC; Janus tyrosine kinase and c-Src kinase are not involved in the RET/PTC-mediated activation of STAT3. RET/PTC-induced activation of STAT3 induces the STAT3 responsive genes, vascular endothelial growth factor, cyclin D1, and intercellular adhesion molecule 1. In addition, RET/PTC-mediated cellular transformation and proliferation of transformed cells require tyrosine 705 phosphorylation of STAT3 in NIH3T3 cells. We conclude that STAT3 activation by the RET/PTC tyrosine kinase is one of the critical signaling pathways for the regulation of specific genes, such as cyclin D1, vascular endothelial growth factor, and intercellular adhesion molecule 1, and for cellular transformation. PMID- 12637587 TI - p190-B RhoGAP regulates mammary ductal morphogenesis. AB - Previous studies from our laboratory have demonstrated that p190-B RhoGAP (p190 B) is differentially expressed in the Cap cells of terminal end buds (TEBs) and poorly differentiated rodent mammary tumors. Based on these observations we hypothesized that p190-B might play an essential role in invasion of the TEBs into the surrounding fat pad during ductal morphogenesis. To test this hypothesis, mammary development was studied in p190-B-deficient mice. A haploinsufficiency phenotype was observed in p190-B heterozygous mice as indicated by decreased number and rate of ductal outgrowth(s) at 3, 4, and 5 wk of age when compared with their wild-type littermates. This appeared to result from decreased proliferation in the Cap cells of the TEBs, a phenotype remarkably similar to that observed previously in IGF-I receptor null mammary epithelium. Furthermore, decreased expression of insulin receptor substrates 1 and 2 were observed in TEBs of p190-B heterozygous mice. These findings are consistent with decreased IGF signaling observed previously in p190-B-/- mouse embryo fibroblasts. To further assess if this defect was cell autonomous or due to systemic endocrine effects, the mammary anlagen from p190-B+/+, p190-B+/-, and p190-B-/- mice was rescued by transplantation into the cleared fat pad of recipient Rag1-/- mice. Surprisingly, as opposed to 75-80% outgrowths observed using wild-type donor epithelium, only 40% of the heterozygous and none of the p190-B-/- epithelial transplants displayed any outgrowths. Together, these results suggest that p190-B regulates ductal morphogenesis, at least in part, by modulating the IGF signaling axis. PMID- 12637588 TI - Selective modulation of genomic and nongenomic androgen responses by androgen receptor ligands. AB - Steroids can induce both transcription-dependent (genomic) and independent (nongenomic) signaling. Here, several classical androgen receptor ligands were tested for their ability to modulate genomic and nongenomic responses, focusing on the role of the oocyte-expressed Xenopus classical androgen receptor (XeAR) in mediating these processes. Cellular fractionation and immunohistochemistry revealed that the XeAR was located throughout oocytes, including within the plasma membrane. RNA interference and oocyte maturation studies suggested that androgen-induced maturation was mediated in part by the XeAR in a transcription independent fashion, perhaps by altering G protein-mediated signaling. While inducing minimal transcription in oocytes, all AR ligands promoted significant XeAR-mediated transcription in CV1 cells. In contrast, only testosterone and androstenedione potently induced oocyte maturation, whereas dihydrotestosterone and R1881 actually inhibited testosterone and human chorionic gonadotropin induced maturation and signaling. These results suggest that the nature of a steroid-induced signal (genomic vs. nongenomic) may depend on the type of target cell, the receptor location within cells, as well as the ligand itself. The identification of molecules capable of selectively altering genomic vs. nongenomic signaling may be useful in delineating the roles of these pathways in mediating androgen responses and might lead to the development of novel compounds that specifically modulate these signals in vivo. PMID- 12637589 TI - Vitamin D: more than a "bone-a-fide" hormone. AB - The vitamin D endocrine system is critical for the proper development and maintenance of mineral ion homeostasis and skeletal integrity. Beyond these classical roles, recent evidence suggests that the bioactive metabolite of vitamin D, 1,25-dihydroxyvitamin D3, functions in diverse physiological processes, such as hair follicle cycling, blood pressure regulation, and mammary gland development. This minireview explores the current progress in unraveling the complexities of the vitamin D endocrine system by focusing on four main areas of research: the resolution of the vitamin D receptor crystal structure, the molecular details of 1,25-dihydroxyvitamin D3-mediated transcription, murine knockout models of key genes in the endocrine system, and alternative vitamin D receptors and ligands. PMID- 12637590 TI - Consensus on evidence or evidence of consensus? The evolving role and the new expertise of the occupational physician. PMID- 12637591 TI - Shift work and occupational medicine: an overview. AB - In modern society, more and more people work during 'non-standard' working hours, including shift and night work, which are recognized risk factors for health, safety and social well-being. Suitable preventive and protective measures are required to mitigate the adverse effects and ensure that the worker can cope satisfactorily. These are based mainly on the organization of shift schedules according to ergonomic criteria and on specific medical surveillance. Occupational medicine has to consider very carefully the several factors (psycho physiological, pathological and social) that can influence tolerance and/or maladaptation. PMID- 12637592 TI - Shift work and disturbed sleep/wakefulness. AB - Of the many health-related effects of shift work, disturbed sleep is the most common. This review describes the main observed effects of the three principal shifts (night, morning and afternoon) on patterns of sleep and wakefulness. The mechanism of sleep disruption in relation to circadian rhythms and the specific impact of aspects of shift organization (speed and direction of rotation) are discussed. The most troublesome acute symptoms are difficulty getting to sleep, shortened sleep and somnolence during working hours that continues into successive days off. These are only partially amenable to amelioration by manipulating shift patterns. However, there is no clear indication that chronic sleep problems result from long-term shift work. PMID- 12637593 TI - Shift work, safety and productivity. AB - The arguments in favour of introducing shift work clearly depend on productivity and safety being maintained at an acceptable level. However, the evidence reviewed in this paper clearly indicates that both productivity and safety may be compromised at night. More specifically, safety declines over successive night shifts, with increasing hours on duty and between successive rest breaks. The only known way to minimize these problems is to improve shift systems with respect to these factors. However, these factors need to be considered in combination with one another since, for example, a long night shift that includes frequent rest breaks might well prove safer than a shorter night shift with less frequent breaks. PMID- 12637594 TI - Health disorders of shift workers. AB - The effects of shift work on physiological function through disruption of circadian rhythms are well described. However, shift work can also be associated with specific pathological disorders. This article reviews the evidence for a relationship between specific medical disorders and working at night or on shift systems. The strongest evidence exists for an association with peptic ulcer disease, coronary heart disease and compromised pregnancy outcome. PMID- 12637595 TI - Preventive and compensatory measures for shift workers. AB - Shift systems are known to be associated with a variety of psychosocial and physiological problems that can affect the health of workers. This review focuses on measures that can be taken to optimize the well-being of shift workers and to identify ill-health at an early stage. The discussion includes specific aspects of the design of shift systems, taking account of variation in the views and circumstances of employees, and strategies to combat sleepiness at work and elsewhere. Although an ideal shift system does not exist, a wholistic approach comprising education of managers, employees and their families can ameliorate some of the health consequences. PMID- 12637596 TI - Audit in occupational medicine: an audit of fitness to drive among voluntary drivers in an NHS trust. AB - BACKGROUND: A health surveillance programme, to assess fitness to drive, was initiated for voluntary drivers in an NHS Trust because of reports of increasing frailty and slow reactions among some drivers. After discussion between the occupational health department, voluntary services manager and personnel department it was considered appropriate to apply Driver and Vehicle Licensing Authority (DVLA) Group 2 fitness to drive standards to those voluntary drivers who drove the Trusts minibuses. RESULTS: An audit of the initial health surveillance of 47 drivers is presented. The mean age of the voluntary drivers was 66.4 years. A large number of medical problems with the potential to affect driving were discovered (average of 1.9 medical problems per driver). The outcome was that five voluntary drivers were found unfit to drive the hospital minibus and one voluntary driver was found unfit for car driving. CONCLUSIONS: A fitness assessment form for drivers is presented. This form is primarily for the use of occupational health nurses, to help them to decide when referral to an occupational health physician is indicated. PMID- 12637597 TI - Neurological effects of radiofrequency radiation. AB - BACKGROUND: The health effects of radiofrequency radiation (RFR) and the adequacy of the safety standards are a subject of debate. One source of human data is case reports regarding peripheral neurological effects of RFR, mainly noxious sensations or dysaesthesiae. AIM: To investigate health effects, neurophysiological mechanisms and safety levels for RFR. METHODS: We conducted a literature search for case reports and case series associated with mobile phone technology as well as other RFR sources using specific search terms on PubMed. RESULTS: We identified 11 original articles detailing case reports or case series and matching the search criteria. Five of the identified papers were written by at least one of the authors (B.H. or R.W.). CONCLUSIONS: Cases have arisen after exposure to much of the radiofrequency range. In some cases, symptoms are transitory but lasting in others. After very high exposures, nerves may be grossly injured. After lower exposures, which may result in dysaesthesia, ordinary nerve conduction studies find no abnormality but current perception threshold studies have found abnormalities. Only a small proportion of similarly exposed people develop symptoms. The role of modulations needs clarification. Some of these observations are not consistent with the prevailing hypothesis that all health effects of RFR arise from thermal mechanisms. PMID- 12637598 TI - Occupational health management: an audit tool. AB - BACKGROUND: Organizations must manage occupational health risks in the workplace and the UK Health & Safety Executive (HSE) has published guidance on successful health and safety management. AIMS: This paper describes a method of using the published guidance to audit the management of occupational health and safety, first at an organizational level and, secondly, to audit an occupational health service provider's role in the management of health risks. The paper outlines the legal framework in the UK for health risk management and describes the development and use of a tool for qualitative auditing of the efficiency, effectiveness and reliability of occupational health service provision within an organization. The audit tool is presented as a question set and the paper concludes with discussion of the strengths and weaknesses of using this tool, and recommendations on its use. PMID- 12637599 TI - Association between lifestyle and white blood cell count: a study of Japanese male office workers. AB - METHODS: We assessed the association of lifestyle factors with white blood cell (WBC) count in 3681 Japanese male office workers aged 35-59 years. RESULTS: The WBC count differed significantly across lifestyle factors (overall obesity, alcohol consumption, cigarette smoking, eating breakfast, nutritional balance, physical exercise and hours of work). From multiple linear regression analyses, overall obesity, alcohol intake, cigarette smoking, nutritional balance and hours of work remained as statistically significant factors associated with WBC count. The WBC count increment showed a positive dose-response relationship with poor lifestyle factors regarding overall obesity, cigarette smoking and nutritional balance. On the other hand, the WBC count increment showed a negative dose response relationship with alcohol intake and hours of work. CONCLUSION: These data indicate that lifestyle factors may exert an important effect on WBC count. PMID- 12637600 TI - Type A lactic acidosis in occupational heat exhaustion. AB - BACKGROUND: This paper presents a further analysis of biochemical data collected during a 1 year prospective study of 106 cases of heat exhaustion at a deep underground metalliferous mine. RESULTS: Multiple regression analysis results indicate that the haemoglobin, serum creatinine and plasma lactate concentrations are statistically significant predictors of the anion gap. Together, they explain 65% of the variance in the anion gap (R(2) = 0.650). Spearman's rho correlation results also confirm that haemoglobin, creatinine and lactate are each statistically significantly correlated with the anion gap (P < 0.001). CONCLUSIONS: These results indicate that dehydration and lactate are important determinants of the metabolic acidosis previously observed in occupational heat exhaustion. It is likely that dehydration in these workers has resulted in poor muscle perfusion, anaerobic conditions and elevated lactate. This constitutes Type A lactic acidosis. Creatine kinase is not a statistically significant predictor of the anion gap in multiple regression (P = 0.956). Furthermore, the Spearman's rho correlation coefficient for creatine kinase versus the anion gap is weak (r(s) = 0.175) and is not statistically significant (P = 0.073). These results suggest that there was no rhabdomyolysis contributing to the metabolic acidosis. PMID- 12637602 TI - Investigation of eye splash and needlestick incidents from an HIV-positive donor on an intensive care unit using root cause analysis. AB - BACKGROUND: Two doctors working on a busy intensive care unit sustained injuries whilst removing a chest drain from an HIV-positive patient. One doctor had a needlestick injury into his finger whilst the other sustained an eyesplash when the chest drain was pulled out. METHODS: Following Department of Health format 'Doing less harm', a root cause and human factor analysis of the incident was carried out. The aim was to explore the underlying issues. RESULTS AND CONCLUSIONS: Training, cultural and organizational issues were exposed, and are now being addressed. This approach has led to a far more effective dialogue with the National Health Trust concerned than was previously experienced, and there is early evidence of progress on important aspects of health and safety management at organizational level. Lack of health and safety training of doctors at undergraduate and postgraduate level needs to be addressed. PMID- 12637601 TI - Elevated salivary cortisol levels as a result of sleep deprivation in a shift worker. AB - BACKGROUND: This paper reports a case from a group of 63 shift-workers for whom adaptation to shift-work was evaluated through measurement of salivary cortisol levels. METHODS: Workers' saliva was sampled and cortisol levels measured at intervals of 2 h during morning, evening and night shifts. RESULTS: For one subject among the 63, very high values of cortisol (an approximately 6-fold increase) were observed for the morning (M) shift, but with normal values found for evening (E) and night (N) shifts. Individual mean and peak cortisol values were 48.4 and 67.8 nmol/l against group mean and peak cortisol values of 8.9 and 11.0 nmol/l. Retrospective questioning showed that this subject was healthy and there were no indicators of long-term stress. CONCLUSION: This cortisol rise was deduced to be caused by sleep deprivation as a result of rapidly rotating shift patterns. PMID- 12637605 TI - Tales of Kieran: the occupational physician's odyssey. PMID- 12637606 TI - Studying peanut anaphylaxis. PMID- 12637607 TI - Factors associated with the development of peanut allergy in childhood. AB - BACKGROUND: The prevalence of peanut allergy appears to have increased in recent decades. Other than a family history of peanut allergy and the presence of atopy, there are no known risk factors. METHODS: We used data from the Avon Longitudinal Study of Parents and Children, a geographically defined cohort study of 13,971 preschool children, to identify those with a convincing history of peanut allergy and the subgroup that reacted to a double-blind peanut challenge. We first prospectively collected data on the whole cohort and then collected detailed information retrospectively by interview from the parents of children with peanut reactions and of children from two groups of controls (a random sample from the cohort and a group of children whose mothers had a history of eczema and who had had eczema themselves in the first six months of life). RESULTS: Forty-nine children had a history of peanut allergy; peanut allergy was confirmed by peanut challenge in 23 of 36 children tested. There was no evidence of prenatal sensitization from the maternal diet, and peanut-specific IgE was not detectable in the cord blood. Peanut allergy was independently associated with intake of soy milk or soy formula (odds ratio, 2.6; 95 percent confidence interval, 1.3 to 5.2), rash over joints and skin creases (odds ratio, 2.6; 95 percent confidence interval, 1.4 to 5.0), and oozing, crusted rash (odds ratio, 5.2; 95 percent confidence interval, 2.7 to 10.2). Analysis of interview data showed a significant independent relation of peanut allergy with the use of skin preparations containing peanut oil (odds ratio, 6.8; 95 percent confidence interval, 1.4 to 32.9). CONCLUSIONS: Sensitization to peanut protein may occur in children through the application of peanut oil to inflamed skin. The association with soy protein could arise from cross-sensitization through common epitopes. Confirmation of these risk factors in future studies could lead to new strategies to prevent sensitization in infants who are at risk for subsequent peanut allergy. PMID- 12637608 TI - Effect of anti-IgE therapy in patients with peanut allergy. AB - BACKGROUND: Peanut-induced anaphylaxis is an IgE-mediated condition that is estimated to affect 1.5 million people and cause 50 to 100 deaths per year in the United States. TNX-901 is a humanized IgG1 monoclonal antibody against IgE that recognizes and masks an epitope in the CH3 region of IgE responsible for binding to the high-affinity Fc(epsilon) receptor on mast cells and basophils. METHODS: We conducted a double-blind, randomized, dose-ranging trial in 84 patients with a history of immediate hypersensitivity to peanut. Hypersensitivity was confirmed and the threshold dose of encapsulated peanut flour established by a double blind, placebo-controlled oral food challenge at screening. Patients were randomly assigned in a 3:1 ratio to receive either TNX-901 (150, 300, or 450 mg) or placebo subcutaneously every four weeks for four doses. The patients underwent a final oral food challenge within two to four weeks after the fourth dose. RESULTS: From a mean base-line threshold of sensitivity of 178 to 436 mg of peanut flour in the various groups, the mean increases in the oral-food-challenge threshold were 710 mg in the placebo group, 913 mg in the group given 150 mg of TNX-901, 1650 mg in the group given 300 mg of TNX-901, and 2627 mg in the group given 450 mg of TNX-901 (P<0.001 for the comparison of the 450-mg dose with placebo, and P for trend with increasing dose <0.001). TNX-901 was well tolerated. CONCLUSIONS: A 450-mg dose of TNX-901 significantly and substantially increased the threshold of sensitivity to peanut on oral food challenge from a level equal to approximately half a peanut (178 mg) to one equal to almost nine peanuts (2805 mg), an effect that should translate into protection against most unintended ingestions of peanuts. PMID- 12637610 TI - Factors associated with progression of carcinoid heart disease. AB - BACKGROUND: By releasing vasoactive substances into the circulation, carcinoid tumors can cause right-sided valvular heart disease. Factors associated with the progression of carcinoid heart disease are poorly understood. We conducted a retrospective study to identify such factors. METHODS: Our sample included 71 patients with the carcinoid syndrome who underwent serial echocardiographic studies performed more than one year apart and 32 patients referred directly for surgical intervention after an initial echocardiographic evaluation. A score for carcinoid heart disease was determined on the basis of an assessment of valvular anatomy and function and the function of the right ventricle. An increase of more than 25 percent in the score between studies was considered suggestive of disease progression. Tumor progression was assessed on the basis of abdominal computed tomographic scans and changes in the level of urinary 5-hydroxyindoleacetic acid (5-HIAA), a metabolite of serotonin. RESULTS: Of the patients with serial echocardiographic studies, 25 (35 percent) had an increase of more than 25 percent in the cardiac score. As compared with patients whose score changed by 25 percent or less, these patients had higher urinary peak 5-HIAA levels (median, 265 mg per 24 hours [interquartile range, 209 to 593] vs. 189 mg per 24 hours [interquartile range, 75 to 286]; P=0.004) and were more likely to have biochemical progression (10 of 25 patients vs. 9 of 46, P=0.05) and to have received chemotherapy (13 of 25 vs. 10 of 46, P=0.009). Logistic-regression analysis showed that a higher peak urinary 5-HIAA level and previous chemotherapy were predictors of an increase in the cardiac score that exceeded 25 percent (odds ratio for each increase in 5-HIAA of 25 mg per 24 hours, 1.08 [95 percent confidence interval, 1.03 to 1.13]; P=0.009); odds ratio associated with chemotherapy, 3.65 [95 percent confidence interval, 1.74 to 7.48]; P=0.001). CONCLUSIONS: Serotonin is related to the progression of carcinoid heart disease, and the risk of progressive heart disease is higher in patients who receive chemotherapy than in those who do not. PMID- 12637609 TI - Imatinib compared with interferon and low-dose cytarabine for newly diagnosed chronic-phase chronic myeloid leukemia. AB - BACKGROUND: Imatinib, a selective inhibitor of the BCR-ABL tyrosine kinase, produces high response rates in patients with chronic-phase chronic myeloid leukemia (CML) who have had no response to interferon alfa. We compared the efficacy of imatinib with that of interferon alfa combined with low-dose cytarabine in newly diagnosed chronic-phase CML. METHODS: We randomly assigned 1106 patients to receive imatinib (553 patients) or interferon alfa plus low-dose cytarabine (553 patients). Crossover to the alternative group was allowed if stringent criteria defining treatment failure or intolerance were met. Patients were evaluated for hematologic and cytogenetic responses, toxic effects, and rates of progression. RESULTS: After a median follow-up of 19 months, the estimated rate of a major cytogenetic response (0 to 35 percent of cells in metaphase positive for the Philadelphia chromosome) at 18 months was 87.1 percent (95 percent confidence interval, 84.1 to 90.0) in the imatinib group and 34.7 percent (95 percent confidence interval, 29.3 to 40.0) in the group given interferon alfa plus cytarabine (P<0.001). The estimated rates of complete cytogenetic response were 76.2 percent (95 percent confidence interval, 72.5 to 79.9) and 14.5 percent (95 percent confidence interval, 10.5 to 18.5), respectively (P<0.001). At 18 months, the estimated rate of freedom from progression to accelerated-phase or blast-crisis CML was 96.7 percent in the imatinib group and 91.5 percent in the combination-therapy group (P<0.001). Imatinib was better tolerated than combination therapy. CONCLUSIONS: In terms of hematologic and cytogenetic responses, tolerability, and the likelihood of progression to accelerated-phase or blast-crisis CML, imatinib was superior to interferon alfa plus low-dose cytarabine as first-line therapy in newly diagnosed chronic-phase CML. PMID- 12637611 TI - Images in clinical medicine. A deadly examination. PMID- 12637612 TI - Shattuck lecture: Diversity of the immune repertoire and immunoregulation. PMID- 12637613 TI - Clinical practice. Vestibular neuritis. PMID- 12637614 TI - Case record of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 8-2003. A 35-year-old man with early dysfunction of a second renal transplant. PMID- 12637615 TI - Two approaches to peanut allergy. PMID- 12637616 TI - Imatinib mesylate--the new gold standard for treatment of chronic myeloid leukemia. PMID- 12637617 TI - Understanding and responding to adverse events. PMID- 12637618 TI - Aspirin and mortality from coronary bypass surgery. PMID- 12637619 TI - C-reactive protein in the prediction of cardiovascular events. PMID- 12637620 TI - Reviparin after leg injury requiring immobilization. PMID- 12637621 TI - Cyclin E in breast cancer. PMID- 12637623 TI - Genetic testing. PMID- 12637622 TI - Skin ulcers misdiagnosed as pyoderma gangrenosum. PMID- 12637624 TI - Delayed-onset heparin-induced thrombocytopenia and cerebral thrombosis after a single administration of unfractionated heparin. PMID- 12637626 TI - Why is coronary heart disease of uraemic patients so frequent and so devastating? AB - On September 6, 2001, Professor Fernando Valderrabano (Hospital Gregorio Maranon, Madrid) died at the age of 59 years. He was a leading figure in Spanish nephrology, a full professor of Medicine/Nephrology at the University Complutense of Madrid, and an outstanding scientist who published more than 300 articles in medical journals. He was a very intelligent and cultured person, and a man of great style who enjoyed a wide range of hobbies and interests in addition to his medical work. All his colleagues and friends mourn his passing. PMID- 12637627 TI - Stem cells of the adult kidney: where are you from? PMID- 12637625 TI - Enfuvirtide, an HIV-1 fusion inhibitor, for drug-resistant HIV infection in North and South America. AB - BACKGROUND: The T-20 vs. Optimized Regimen Only Study 1 (TORO 1) was a randomized, open-label, phase 3 study of enfuvirtide (T-20), a human immunodeficiency virus type 1 (HIV-1) fusion inhibitor. METHODS: Patients from 48 sites in the United States, Canada, Mexico, and Brazil with at least six months of previous treatment with agents in three classes of antiretroviral drugs, resistance to drugs in these classes, or both, and with at least 5000 copies of HIV-1 RNA per milliliter of plasma were randomly assigned in a 2:1 ratio to receive enfuvirtide plus an optimized background regimen of three to five antiretroviral drugs or such a regimen alone (control group). The primary efficacy end point was the change in the plasma HIV-1 RNA level from base line to week 24. RESULTS: A total of 501 patients underwent randomization, and 491 received at least one dose of study drug and had at least one measurement of plasma HIV-1 RNA after treatment began. The two groups were balanced in terms of the median base-line HIV-1 RNA level (5.2 log10 copies per milliliter in both groups), median CD4+ cell count (75.5 cells per cubic millimeter in the enfuvirtide group, and 87.0 cells per cubic millimeter in the control group), demographic characteristics, and previous antiretroviral therapy. At 24 weeks, the least-squares mean change from base line in the viral load (intention-to treat, last observation carried forward) was a decrease of 1.696 log10 copies per milliliter in the enfuvirtide group, and a decrease of 0.764 log10 copies per milliliter in the control group (P<0.001). The mean increases in CD4+ cell count were 76 cells per cubic millimeter and 32 cells per cubic millimeter, respectively (P<0.001). Reactions at the site of the injections were reported by 98 percent of patients receiving enfuvirtide. There were more cases of pneumonia in the enfuvirtide group than in the control group. CONCLUSIONS: The addition of enfuvirtide to an optimized antiretroviral regimen provided significant antiretroviral and immunologic benefit through 24 weeks in patients who had previously received multiple antiretroviral drugs and had multidrug-resistant HIV 1 infection. PMID- 12637628 TI - Protocol biopsy: what is the rationale and what is the evidence? PMID- 12637629 TI - Can technology solve the clinical problem of 'dry weight'? PMID- 12637630 TI - High permeability of dialysis membranes: what is the limit of albumin loss? PMID- 12637631 TI - Transplant capillaropathy and transplant glomerulopathy: ultrastructural markers of chronic renal allograft rejection. PMID- 12637632 TI - Renal replacement therapy in Bosnia and Herzegovina: report of the Society of Nephrology, Dialysis and Transplantation of Bosnia and Herzegovina. PMID- 12637633 TI - Increased expression of monocyte chemoattractant protein-1 (MCP-1) by renal epithelial cells in culture on exposure to calcium oxalate, phosphate and uric acid crystals. AB - BACKGROUND: During the development of non-infectious kidney stones, crystals form and deposit in the kidneys and become surrounded by monocytes/macrophages (M/M). We have proposed that in response to crystal exposure renal epithelial cells produce chemokines, which attract the M/M to the sites of crystal deposition. We investigated the expression of monocyte chemoattractant protein-1 (MCP-1) mRNA and protein by NRK52E rat renal tubular epithelial cells exposed to calcium oxalate (CaOx), brushite (Br, a calcium phosphate) and uric acid (UA) crystals. METHODS: Confluent cultures of NRK52E cells were exposed to CaOx, Br or UA at a concentration of 250 micro g/ml (66.7 micro g/cm(2)). They were exposed for 1, 3, 6, 12, 24 and 48 h for isolation of mRNA and 24 h for ELISA to determine the secretion of protein into the culture medium. Since cells are known to produce free radicals on exposure to CaOx crystals we also investigated the effect of free radical scavenger catalase on the crystal induced expression of MCP-1 mRNA and protein. RESULTS: Exposure of NRK52E cells to the crystals resulted in increased expression of MCP-1 mRNA and production of the chemoattractant. CaOx crystals were most provocative while UA the least. Treatment with catalase had a negative effect on the increased expression of both MCP-1 mRNA and protein, which indicates the involvement of free radicals in up-regulation of MCP-1 production. CONCLUSION: Exposure to both CaOx and calcium phosphate crystals stimulates increased production of MCP-1. Free radicals appear to be involved in this up regulation. Results indicate that MCP-1, which is often associated with localized inflammation, may be one of the chemokine mediators associated with the deposition of various urinary crystals in the kidneys during kidney stone formation. Because of the small number of experiments performed here, results must be confirmed by more extensive studies with larger sample size. PMID- 12637634 TI - Pentoxifylline modulates intracellular signalling of TGF-beta in cultured human peritoneal mesothelial cells: implications for prevention of encapsulating peritoneal sclerosis. AB - BACKGROUND: Peritoneal matrix accumulation is a major characteristic of encapsulating peritoneal sclerosis (EPS), which is a serious complication in long term peritoneal dialysis (PD) patients. We reported previously that TGF-beta stimulates collagen gene expression in cultured HPMC, and is attenuated by pentoxifylline (PTX). The SMAD family and the mitogen-activated protein kinase (MAPK) (ERK1/2, JNK and p38(HOG)) pathways have been shown to participate in TGF beta signalling. However, how PTX modulates the intracellular signalling downstream to TGF-beta remains undetermined in HPMC. In this study, we explored these signalling pathways in HPMC, and investigated the molecular mechanisms involved in the inhibitory effects of PTX on TGF-beta-induced collagen gene expression in HPMC. METHODS: HPMC was cultured from human omentum by an enzyme digestion method. The expression of collagen alpha1(I) mRNA was determined by northern blotting, while the SMAD proteins and the MAPK kinase activity were determined by western blotting. RESULTS: TGF-beta-stimulated collagen alpha1(I) mRNA expression of HPMC was inhibited by PTX. The Smad2, ERK1/2 and p38(HOG) pathways were activated in response to TGF-beta. However, TGF-beta displayed no activation of the JNK pathway in HPMC. The addition of PD98059 and SB203580, which blocked the activation of ERK1/2 and p38(HOG), respectively, suppressed the TGF-beta-induced collagen alpha1(I) mRNA expression. At a concentration (300 micro g/ml) that inhibited the collagen gene expression, PTX suppressed the ERK1/2 and p38(HOG) activation by TGF-beta. In contrast, PTX had no effect on the TGF-beta-induced activation of Smad2, under the same concentration. CONCLUSION: PTX inhibits the TGF-beta-induced collagen gene expression in HPMC through modulating the ERK1/2 and p38(HOG) pathways. Our study of PTX may provide the therapeutic basis for clinical applications in the prevention of EPS. PMID- 12637635 TI - Glucose degradation products increase apoptosis of human mesothelial cells. AB - BACKGROUND: The heat sterilization of glucose solutions for peritoneal dialysis (PDS) induces the formation of glucose degradation products (GDPs), a phenomenon amplified by lactate and neutral pH. In the new three-compartment bag (3CB) PDS, a glucose solution at pH 3 is kept apart from the buffer until use, and the final solution delivers glucose concentrations that are similar to traditional PDS (TPD), with pH 6 and a lower content of GDPs. As GDPs have oxidant activity that may favour apoptosis, we investigated mesothelial cell apoptosis modulation by 12 h cultures in media supplemented with: (i) two relevant GDPs, methylglyoxal (MGly) and formaldehyde (For) in time and dose-dependence assays, (ii) GDPs at concentrations detected in TPD and 3CB, and (iii) commercial TPD and 3CB PDS, both with 1.36% glucose. METHODS: Apoptosis was evaluated by terminal 3' uridine labelling. Key proteins involved in the apoptotic pathway were investigated by reverse transcription polymerase chain reaction (RT-PCR) mRNA expression and immunoperoxidase staining (caspase 9, tumour suppressor protein p53, inducible cyclooxygenase COX-2). RESULTS: The apoptotic effects of MGly and For were dose and time dependent. GPDs at concentrations detected in TPD induced greater transcription and translation of apoptotic pathway proteins (caspase 9, p53 and COX-2) than GPDs in 3CB. This resulted in a higher apoptotic rate, which was not influenced by addition of sterile glucose. A similar enhancement of apoptosis was detected when mesothelial cells were incubated with TPD, whereas incubation in 3CB PDS resulted in less enhanced apoptosis. The 12 h incubation effect of PDS on cultured mesothelial cells was not related to advanced glycosylated end-product formation. CONCLUSIONS: As the rate of mesothelial cell apoptosis is lower in 3CB than in TPD solutions, the 3CB appears to provide improved biocompatibility. PMID- 12637636 TI - Nephrotic urine prevents increased rat glomerular albumin permeability induced by serum from the same patient with idiopathic nephrotic syndrome. AB - BACKGROUND: The putative humoral mediator thought to be involved in the pathogenesis of idiopathic nephrotic syndrome has not yet been identified. However, components exist in normal serum that block the permeability activity of FSGS serum in vitro. The potential of FSGS serum to increase glomerular albumin permeability may result from an imbalance between permeability factors and naturally occurring inhibitors. We hypothesized that this imbalance may be favoured by loss of inhibitory factors in nephrotic urine. METHODS: The study population consisted of seven patients with biopsy-proven FSGS, one with IgM nephropathy, and three with idiopathic nephrotic syndrome without biopsy, from whom frozen serum and dialysed and lyophilized urine samples were available. Glomerular albumin permeability (P(alb)) was determined from the change in glomerular volume induced by applying oncotic gradients across the basement membrane of normal isolated rat glomeruli pre-incubated with patient serum, normal control serum, patient serum mixed with an equal volume of urine from the same patient, or patient serum mixed with normal urine. Serum and urine apolipoproteins J and E were measured by dot-blot, utilizing peroxidase-labelled antibodies. The urinary capacity to scavenge oxygen radicals was determined after exposure of isolated glomeruli to superoxide generated by xanthine and xanthine oxidase. RESULTS: The mean P(alb) of the patients was markedly elevated at 0.74+/ 0.08. The addition of urine from the same patient significantly reduced P(alb) (mean 0.15+/-0.23) in all but one of the patients with FSGS. Normal urine had no inhibitory effect in the 10 patients in which it was tested (mean 0.71+/-0.09). Serum apo J was slightly decreased and serum apo E was slightly increased compared with controls. Urine levels of both lipoproteins were significantly decreased compared with controls. Urine from FSGS patients effectively neutralized superoxide, whereas normal urine did not. CONCLUSIONS: Nephrotic urine but not normal urine contains components that block increased albumin permeability in isolated rat glomeruli induced by serum from patients with the idiopathic nephrotic syndrome. The inhibitory function of these components, which appear not to include apolipoproteins J and E, may involve scavenging of superoxide as a final common pathway. Loss in the urine from the serum of naturally occurring inhibitors in the initial stages of the disease may propagate proteinuria and glomerular injury. PMID- 12637637 TI - The effect of growth hormone on the development of diabetic kidney disease in rats. AB - BACKGROUND: Nephropathy is the most severe complication of diabetes mellitus. We investigated the effect of exogenous growth hormone (GH) administration on renal function and matrix deposition in the streptozotocin (STZ) model of type I diabetic rat. METHODS: Adult female STZ-diabetic rats (D), non-diabetic control rats injected with saline (C) and control and diabetic rats injected with bovine GH for 3 months (CGH and DGH, respectively) were used. RESULTS: The usual renal hypertrophy seen in D animals was more pronounced in the DGH group. Creatinine clearance increased only in the D rats, but not in the other groups, including DGH. Albuminuria was observed in the D animals but was significantly elevated in the DGH group. Glomeruli from DGH animals showed more extensive matrix accumulation (manifested as an increase in mesangial/glomerular area ratio). Renal extractable insulin-like growth factor (IGF-I) mRNA was decreased in the D and DGH groups, but renal IGF-I protein was not significantly increased. Renal IGF binding protein-1 was increased in the D groups and further increased in the DGH group, at both the mRNA and protein levels. CONCLUSIONS: GH-treated diabetic rats had less hyperfiltration and more albuminuria, concomitant with more glomerular matrix deposition, when compared with regular diabetic animals. This was associated with a significant increase in renal IGFBP-1, and dissociated from IGF-I changes. Thus, in this model, GH exacerbates the course of diabetic kidney disease. PMID- 12637638 TI - Lipid-lowering-independent effects of simvastatin on the kidney in experimental hypercholesterolaemia. AB - BACKGROUND: Hypercholesterolaemia (HC), an independent risk factor for renal injury, is associated with formation of oxidized low-density-lipoprotein (ox LDL), increased oxidative-stress and renal inflammation. HMG-CoA-reductase inhibitors are commonly used in HC, but their effects on renal haemodynamics and function in HC are poorly understood. METHODS: Pigs were studied after a 12-week normal diet, a 2% high-cholesterol diet (HC) or an HC diet supplemented with simvastatin (HC+simvastatin, 80 mg/day) (n=6-8 each group). Renal haemodynamics and function were quantified in vivo with electron-beam computed tomography (EBCT). Shock-frozen renal tissue was subsequently studied using immunohistochemistry. RESULTS: LDL cholesterol was similarly increased in HC and HC+simvastatin. Simvastatin-treated animals showed increased expression of endothelial nitric-oxide-synthase (eNOS), and decreased expression of the ox-LDL receptor LOX-1 in renal endothelial cells. Simvastatin also decreased tubular immunoreactivity of inducible-NOS, nitrotyrosine, nuclear-factor-kappaB, and tubuloglomerular trichrome staining. These were associated with a significant increase in cortical (6.1+/-0.1 vs 5.0+/-0.3 and 5.0+/-0.1 ml/min/cc, respectively, P<0.001) and medullary perfusion in HC+simvastatin compared to normal and HC. CONCLUSIONS: Simvastatin attenuated the inflammatory and pro oxidative environment as well as fibrosis in kidneys in pigs with diet-induced HC, in association with enhanced renal perfusion. These cholesterol-lowering independent changes imply novel renoprotective effects of statins in the setting of HC and atherosclerosis. PMID- 12637639 TI - A novel evaluation method for paraffinized human renal biopsies using quantitative analysis of microdissected glomeruli and VCAM-1 as marker of inflammatory mesangial cell activation. AB - BACKGROUND: In the glomerular mesangium, immunologic and/or infectious activation of the inflammatory, NF-kappaB-mediated signal pathway can induce a progression of already existing mesangial lesions in non-immunologic and immunologic glomerular disease. This progression is preceded by upregulated mesangial gene expression of which the vascular cell adhesion molecule-1, VCAM-1 (vascular cell adhesion molecule-1), is a well-established marker. Its evaluation on minimal tissue such as routinely paraffinized needle core biopsies is not established and needs the development of a novel evaluation method more meaningful than common immunohistology. METHODS: By laser-microdissection, 10 glomeruli/case were isolated from 5 micro m thick tissue slices in a total of 15 cases of mesangial proliferation with different renal diseases (IgA nephropathy, lupus nephritis and mesangial proliferative lesions of unknown aetiology) vs transplant biopsies as negative and TNF alpha-treated cultured human mesangial cells as positive controls. After reverse transcription of isolated RNA, cDNA aliquots were quantified for VCAM-1 expression by real-time PCR using the threshold cycle (C(t)) method, normalized for the housekeeping gene beta-actin, and compared with qualitative RT-PCR results. RESULTS: Unsuspected VCAM-1 transcript steady-state levels could be detected by real-time PCR in agreement with qualitative PCR, while morphologic and immunohistologic analyses were unrevealing. As yields of RNA extraction in femtogram quantities cannot be measured spectrophotometrically, a C(t)-ratio was formed between beta-actin and VCAM-1 per case showing high VCAM 1 expression in lupus nephritis (1.39), and moderate expression in IgA nephropathy (1.08-1.23) vs TNF alpha-treated mesangial cells (0.97-1.23) and negative control cases (0.66-0.68). CONCLUSIONS: This is the first reported gene expression analysis method for routinely paraffininzed human renal biopsies, demonstrating the power of combined laser-microdissection and PCR quantification as novel methods for the evaluation of minimal tissue beyond purely descriptive morphologic analysis. PMID- 12637640 TI - Novel truncating mutations in the ClC-5 chloride channel gene in patients with Dent's disease. AB - BACKGROUND: Dent's disease is characterized by low-molecular-weight proteinuria, hypercalciuria, nephrocalcinosis, nephrolithiasis, rickets and eventual renal failure. The disease is caused by mutations in the X-linked chloride channel CLCN5 gene, which encodes a 746-amino-acid protein expressed in renal tubules. These mutations have been reported in unrelated families from the UK, USA, Japan and other countries. We were interested in identifying additional mutations in the CLCN5 coding region of Spanish patients with Dent's disease. METHODS: Five patients from three unrelated Spanish families were studied. Leukocyte genomic DNA from patients and their relatives was used with CLCN5-specific primers for polymerase chain reaction amplification of the coding region and exon-intron boundaries. Amplified products were analysed by single-strand conformational polymorphism analysis, DNA sequencing and restriction enzyme analysis. RESULTS: Low-molecular-weight proteinuria and hypercalciuria were detected in all the patients, nephrocalcinosis in two patients, and rickets or osteopenia in three patients. We identified three new CLCN5 mutations consisting of two nonsense mutations, Leu433Stop and Arg718Stop, and an insertional frameshift mutation, 65insT, which results in a stop at codon 98. These three mutations predict truncated ClC-5 proteins that, respectively, lack 314, 649 and 28 amino acids at the carboxy terminus, and are likely to result in loss of function. These mutations were shown to co-segregate with the disease in each of the three families. CONCLUSIONS: Our study is the first to characterize mutations in the CLCN5 gene in Spanish patients with Dent's disease and expands the spectrum of CLCN5 mutations associated with this disease. PMID- 12637641 TI - Polymorphisms of the tumour necrosis factor alpha gene at position -308 and TNFd microsatellite in primary IgA nephropathy. AB - BACKGROUND: The development of glomerular inflammation in immunoglobulin A nephropathy (IgAN) has been associated with various cytokines, including tumour necrosis factor alpha (TNFalpha). A biallelic polymorphism in the promoter region of the TNFalpha gene (TNFA), at position -308, has been described (TNFA-1 and TNFA-2) and is associated with increased TNFalpha production for the TNFA-2 allele. Another microsatellite polymorphism has been described for TNFd, which is functional and associated with increased production of TNFalpha for the d3 allele. METHODS: We have studied these two polymorphisms in 242 Caucasian patients with biopsy-proven IgAN (169 male, 73 female), who were followed from 1990 to 1999, and in 210 appropriate local Caucasian controls (133 male, 77 female) for comparison of genotypes and allelic distribution. RESULTS: The respective frequencies of A1/A1, A1/A2 and A2/A2 TNFA genotypes were 76.4, 22.3 and 1.3% in IgAN vs 78.1, 19.5 and 2.4% in controls (P=NS). For TNFd, the frequencies of the respective genotypes d3/d3, d3/non-d3 and non-d3/non-d3 were significantly different (chi(2)=12.30, P=0.002, Pc=0.013) with an increased frequency of the low-producer genotype non-d3/non-d3 in IgAN patients (24 vs 12%). The combination of TNFA and TNFd polymorphisms demonstrated that compared with controls, patients with non-A2 and non-d3 alleles (low producers) were more common (18 vs 9%; P=0.006). In the genotype/clinical phenotype correlations, we could not demonstrate significant differences between the different subgroups of patients. However, high-producer TNFalpha patients (A2 and d3 alleles) had more chronic renal failure than others (36.6 vs 22.9%) at last follow-up and their survival without chronic renal failure (Kaplan-Meier) was lower. Nevertheless, TNFalpha polymorphisms were not an independent risk factor for the progression of the disease. CONCLUSIONS: TNFA and TNFd polymorphisms seem to influence the occurrence or initiation of the disease, but do not play a significant role, if any, in the progression of IgA nephritis. PMID- 12637643 TI - Cardiac troponin T and malondialdehyde modified plasma lipids in haemodialysis patients. AB - BACKGROUND: In patients with end-stage renal disease (ESRD), treated with haemodialysis, a high overall mortality is observed. A previous study showed that cardiac troponin T (cTnT) is a strong independent predictor of outcome in this population. In this study we investigated possible causes of cTnT increase and its relationship with a marker of oxidative stress. METHODS: In a group of 71 haemodialysis patients (36 male, 35 female, mean age 68.7+/-1.5 years) we determined cTnT and compared its presence with several biochemical parameters and with malondialdehyde (MDA), which is an indicator of oxidative stress. None of the patients suffered an acute coronary event during the observation period. Three measurements of cTnT and MDA were performed with a 2-week interval. Forty nine patients underwent a transthoracic echocardiography. RESULTS: Twenty-nine patients (or 40.8%) had a positive cTnT determination (defined as cTnT >/=0.10 ng/ml). cTnT positive patients had significantly higher levels of MDA (P=0.0125), C-reactive protein (CRP) (P=0.04) and pre-dialysis urea (P=0.04). Regression analysis showed that both pre-dialysis urea and MDA independently influenced cTnT. No correlation was found with age, dialysis adequacy, post-dialysis urea, total cholesterol, white blood cell count, fibrinogen or any of the echocardiographical parameters. Presence of heart failure, diabetes or use of medication could not discriminate between cTnT positive and cTnT negative patients. MDA levels correlated positively with time on haemodialysis (P=0.0021). Echocardiography showed left ventricular hypertrophy in 88% of the examined patients and impaired wall motion in 35%. Patients with clinical signs of heart failure had a lower ejection fraction and worse wall motion score index. No correlation existed between echocardiographic findings and cTnT or MDA. Survival was independently predicted by cTnT (P=0.0025), MDA (P=0.0007), CRP (P=0.006) and age (P=0.0143). Patients with both cTnT and CRP increase had a survival of <50% at 1 year, compared with 90% in patients with both cTnT and CRP within the normal range and 80% when either CRP or cTnT was increased (chi(2)=12.127; P=0.0023). CONCLUSIONS: This study confirms that the presence of cTnT predicts prognosis in ESRD. The presence of cTnT is linked to oxidative stress, inflammation and uraemia. The absence of specific findings on EKG and echocardiography points towards subclinical myocardial damage caused by endothelial disturbances. PMID- 12637642 TI - Outcome in a post-cardiac surgery population with acute renal failure requiring dialysis: does age make a difference? AB - BACKGROUND: Acute renal failure (ARF), requiring dialysis (ARF-d), develops in 1 5% of patients undergoing cardiac surgery and is associated with higher in hospital mortality. Age is one of the known risk factors for the development of ARF. As the ageing population is increasing, the nephrologist will be faced with a large population of elderly patients requiring dialysis following cardiac surgery. The aim of our study was to evaluate the influence of age on and the risk factors for in-hospital mortality. METHODS: Eighty-two patients with ARF following cardiac surgery and requiring dialysis between January 1997 and October 2001 were included. Two groups of patients were studied: the younger population (<70 years, 42 patients, mean age 59+/-10) and an elderly population (>/=70 years, 40 patients, mean age 76+/-4). Severity of disease was evaluated using the SAPS (Simplified Acute Physiology Score), the Liano score and the SHARF (Stuivenberg Hospital Acute Renal Failure) score. RESULTS: Overall mortality in the population with ARF-d was 56.1%. No difference in mortality rate was found between the younger (61.9%) and elderly patient group (50.0%). The two groups were very similar in baseline and procedural characteristics with exception of body weight (P=0.02) and preoperative glomerular filtration rate (P=0.0001). No significant difference was found in the scoring systems between the old and the young (SAPS P=0.52; Liano P=0.96; SHARF T0 P=0.06; SHARF T48 P=0.15). Mortality in the elderly was significantly correlated with hypotension before starting renal replacement therapy (RRT) (P=0.002), mechanical ventilation (P=0.002), presence of multiorgan failure (MOF) (P=0.0001) and higher scores in the severity models (SAPS: P=0.01; Liano: P<0.0001 and SHARF: P<0.0001). CONCLUSION: The outcome in the elderly requiring dialysis due to ARF post-cardiac surgery is comparable with the outcome in a younger population. No significant difference was found in severity of disease between the elderly and the younger. Variables predicting mortality in the elderly are the presence of MOF, mechanical ventilation and hypotension 24 h before starting RRT. These findings indicate that at the time the nephrologist is called for an elderly patient requiring dialysis due to ARF following cardiac surgery, age per se is not a reason to withhold RRT. PMID- 12637644 TI - Effects of calcitriol on parathyroid function and on bone remodelling in secondary hyperparathyroidism. AB - BACKGROUND: Secondary hyperparathyroidism (2HPT) develops in chronic renal failure due to disturbances of calcium, phosphorus and vitamin D metabolism. It is characterized by high turnover bone disease and an altered calcium-parathyroid hormone (PTH) relationship. Calcitriol has been widely used for the treatment of 2HPT. However, it remains controversial whether calcitriol is capable of inducing changes of the calcium-PTH curve. The aim of the present study was to examine this issue and to determine the effect of calcitriol on bone remodelling in patients with severe 2HPT. METHODS: We evaluated 16 chronic haemodialysis patients with severe 2HPT (PTH 899+/-342 pg/ml). Each patient underwent a dynamic parathyroid function test (by infusion of calcium gluconate and sodium citrate) and a bone biopsy before and after a 6 month period of i.v. calcitriol therapy (CTx). RESULTS: After treatment, eight patients were identified as calcitriol responders and the other eight as non-responders, based on plasma PTH level (<300 pg/ml for responders and >300 pg/ml for non-responders). The first group had higher plasma 25OHD(3) levels (39+/-8 vs 24+/-7 ng/ml, P<0.005). As to the calcium-PTH curve, we found differences in slope (-12.7+/-5.2 vs -21.7+/-11.4, P=0.05), basal/maximum PTH ratio (48.8+/-14.9 vs 71.05+/-20.1%, P=0.01) and time to achieve hypocalcaemia (79.0+/-13.5 vs 94.3+/-13.7 min, P<0.001). Initial histomorphometric parameters did not allow identification of the different groups. After the 6-month CTx, alterations in the calcium-PTH curve were clearly seen in responders [a drop in maximum PTH (from 1651+/-616 to 938+/-744 pg/ml, P<0.05) and minimum PTH (from 163+/-75.4 to 102.2+/-56.7 pg/ml, P<0.005)], associated with an increase in minimum/basal PTH ratio (from 23.3+/-11.6 to 34.5+/-20.4%, P<0.05) and maximum calcium (from 0.99+/-0.07 to 1.1+/-0.09 mmol/l, P<0.05). Set point and slope were not altered after calcitriol treatment, in responders (set point=1.17+/-0.08 vs 1.15+/-0.1 mmol/l, ns; slope=-12.7+/-5.2 vs 12.9+/-9.3, ns) or non-responders (set point=1.21+/-0.05 vs 1.21+/-0.2 mmol/l, ns; slope=-21.7+/-11.4 vs -17.3+/-8.4, ns). Bone formation parameters were reduced in all patients [osteoid surface (OS/BS)=from 57.1+/-21.6 to 41.6+/-26%, P<0.05 for responders, and from 76.7+/-12 to 47.1+/-15%, P<0.001 in non responders], but non-responders had increased bone resorption [eroded surface (ES/BS)=7.1+/-3.4 vs 16.6+/-4.9, P<0.05]. CONCLUSION: Calcitriol had non-uniform effects on parathyroid function and bone remodelling in uraemic patients. Non responders exhibited a decoupled remodelling process that could further influence mineral balance or possibly also bone structure. To avoid such undesirable effects, early identification of non-responder patients is crucial when using calcitriol for the treatment of 2HPT. PMID- 12637646 TI - Influence of PTH assay methodology on differential diagnosis of renal bone disease. AB - BACKGROUND: Determination of plasma parathyroid hormone (PTH) is routinely performed to diagnose and monitor renal bone disease. Recently, a new PTH assay ('whole PTH') using an antibody directed specifically against PTH(1-4) has been introduced. It was the aim of the current study to evaluate whole PTH and parameters derived from whole PTH in renal bone disease. METHODS: The following measurements were carried out in blood samples from 141 unselected haemodialysis patients: three intact PTH assays (Nichols, Roche Elecsys), Scantibodies total); whole PTH (Scantibodies); bone-specific alkaline phosphatase (bAP); tartrate resistant acid-phosphatase 5b (TRAP 5b); osteocalcin, 25-hydroxyvitamin D. Parameters derived from whole PTH were: (i) non-PTH(1-84), difference between intact PTH (Scantibodies assay) and whole PTH; (ii) whole PTH/non-PTH(1-84) ratio. RESULTS: The values generated by the intact PTH assays were comparable. The mean whole PTH concentration was lower than mean intact PTH concentrations (16.9+/-18.1 vs 26.4+/-30.5 pmol/l, Nichols, P<0.05). The correlation coefficients between all four PTH assays were comparable and were very high (r>0.96, ns). The rank order of values generated by the whole PTH assay was statistically not significantly different from the rank order generated by the Nichols intact PTH assay. The median non-PTH(1-84) concentration was 5.2 pmol/l (range 0-49.4). All PTH assays correlated highly significantly with non-PTH(1-84) (correlation coefficients 0.83-0.92). Corrected serum calcium was also associated with non-PTH(1-84) but the correlation was weaker (r=0.28). Regression analysis indicated that the non-PTH(1-84) concentration could be predicted by 76.6-84.6% by the prevailing intact PTH concentrations. Other parameters contributed only marginally to prediction of non-PTH(1-84). In the entire patient group, there was no statistically significant correlation between the whole PTH/non-PTH(1-84) ratio and any of the PTH assays or biochemical bone markers. Eight of 141 patients had a whole PTH/non-PTH(1-84) ratio <1. TRAP 5b, bAP and osteocalcin had high correlations with intact PTH assays and the whole PTH assay (correlation coefficients 0.51-0.56, no significant difference). None of the PTH assays was superior to any other PTH assay in predicting serum concentrations of the bone markers. Therapy with active vitamin D metabolites (n=70) did not alter the results of our analyses. CONCLUSIONS: With respect to information about bone turnover we were not able to find differences between whole PTH and intact PTH assays. Our data also suggest that whole PTH and intact PTH assays give similar information. (i) The correlation between all PTH assays was very high. (ii) The rank order between whole PTH and Nichols intact PTH assays was comparable. (iii) The association between intact PTH assays and non-PTH(1-84) was very high. Albeit non-PTH(1-84) was mostly determined by the prevailing intact PTH concentration, diagnostic information on parathyroid activity provided by whole PTH or intact PTH, respectively, may differ in individual patients. How often this would happen cannot be answered with the currently available data. Unequivocal structural identification of the non-PTH(1-84) fraction would facilitate the answer to that question. The use of the whole PTH/non-PTH(1-84) ratio as a biochemical bone marker in renal bone disease requires further investigation. PMID- 12637645 TI - Pharmacokinetics of doxercalciferol, a new vitamin D analogue that lowers parathyroid hormone. AB - BACKGROUND: This is the first detailed pharmacokinetic report published on the administration of doxercalciferol [1alpha(OH)D(2)] recently introduced to treat secondary hyperparathyroidism. METHODS: 1alpha(OH)D(2) was administered in a range of single and multiple doses to volunteers with and without normal renal and/or hepatic function. Subsequent serial blood samples were assayed by HPLC/radioimmunoassay for the metabolite 1,25-dihydroxyvitamin D(2) [1,25(OH)(2)D(2)], the major active species. RESULTS: Bioavailability of 1,25(OH)(2)D(2) from a single 5 micro g 1alpha(OH)D(2) oral-capsule dose was estimated to be normally approximately 42% of that from a 5 micro g intravenous injection. Steady-state serum concentrations of 1,25(OH)(2)D(2) were attainable within 8 day, and fluctuated approximately 2.5-fold from peak to trough when oral 1alpha(OH)D(2) doses were taken every second day, and the terminal half-life was 34+/-14 h. Mean steady-state serum concentrations rose less than proportionally (from 20 to 45 pg/ml) on increasing oral 1alpha(OH)D(2) doses from 5 to 15 micro g every 48 h. Renal patients showed 39+/-37% increase in serum 1,25(OH)(2)D(2) concentration during 3-4 h haemodialysis sessions, but no other difference in steady-state pharmacokinetics was found between these or hepatically impaired patients and normal subjects. CONCLUSIONS: Given the sensitivity limits of current assays, the pharmacokinetics of this and other vitamin-D compounds is best elucidated from steady-state studies. The pharmacokinetics of 1,25(OH)(2)D(2) from 1alpha(OH)D(2) doses appears to be similar to that of 1,25(OH)(2)D(3) from 1alpha(OH)D(3) doses, albeit D(3) data have to date largely derived from single-dose studies. Deviation of 1,25(OH)(2)D(2) pharmacokinetics from linearity appears to be marginal enough to be clinically manageable with adequate precaution. PMID- 12637647 TI - Kinetics and dosing predictions for daily haemofiltration. AB - BACKGROUND: Thrice-weekly haemofiltration affords excellent outcome when it is used to treat chronic renal failure patients. Daily haemofiltration (DHF) has recently been proposed as a more intensive therapy option, but the total ultrafiltration or exchange volume (replacement volume plus net ultrafiltration volume) requirements for adequate solute clearances during this novel therapy are unknown. METHODS: We calculated theoretical solute kinetic profiles during six times per week DHF for comparison with those during thrice-weekly haemodialysis using a high-flux dialyser (HFHD) or during continuous ambulatory peritoneal dialysis (CAPD). HFHD and CAPD were chosen for comparison because K/DOQI guidelines have defined adequate treatment doses for these therapies. Steady state concentrations were calculated using a two-compartment model of an anuric patient with 35 l of total body water for five solutes: urea, creatinine, vitamin B(12), inulin and beta(2)-microglobulin. Solute distribution volumes and generation rates were taken from the literature, and excess fluid (1 l/day) was assumed to accumulate in and be removed from the extracellular fluid compartment. Theoretical predictions of solute clearance were compared for a 15-l exchange volume/session during DHF, urea Kt/V of 3.6/week during HFHD and urea Kt/V of 2.0/week during CAPD as solute-specific values of the equivalent renal clearance (EKR) and standard Kt/V (stdKt/V) recently defined by Gotch. Additional comparisons of solute clearances were performed between DHF and other daily therapies including six times per week short daily haemodialysis (SDHD) and six times per week nocturnal haemodialysis (NHD). RESULTS: The calculated results predict that: (i) urea clearance during DHF with an exchange volume of 90 l/week (6x15 l) is equivalent to those during HFHD and CAPD based on urea stdKt/V; and (ii) middle molecule clearances during DHF exceed those achieved during HFHD and CAPD based on either EKR or stdKt/V. As expected, DHF therapy was inferior regarding the clearance of urea and other small solutes to SDHD and NHD; however, DHF therapy was superior to SDHD regarding the clearance of larger middle molecules, approaching the clearances achieved by NHD. CONCLUSIONS: We predict that an exchange volume of approximately 40% of total body water (15/35 l=43%) per session will provide adequate clearance of small solutes and substantial clearance of middle molecules during six times per week DHF therapy. These theoretical predictions require clinical validation. PMID- 12637648 TI - Influenza vaccination of dialysis patients: cross-reactivity of induced haemagglutination-inhibiting antibodies to H3N2 subtype antigenic variants is comparable with the response of naturally infected young healthy adults. AB - BACKGROUND: Annual influenza vaccination is recommended for patients with chronic renal failure, although vaccination responses in haemodialysis (HD) patients may be suboptimal. Typically, the seroreactivity has been analysed against the vaccine virus or the corresponding year's epidemic virus. No studies analysing cross-reactivity against subsequent years' viruses have been presented. METHODS: Twenty-three chronic HD patients and 26 cardiac patients were, in autumn 1995, vaccinated with a trivalent influenza vaccine. The cross-reacting haemagglutination-inhibiting antibodies to five consecutive years' (the last season 1999-2000) drift variants of H3N2 subtype influenza A virus were measured and compared with those of vaccinated cardiac patients and with those of 26 healthy military conscripts who suffered a serologically confirmed influenza A infection in the season 1995-1996. RESULTS: The influenza vaccination in HD patients resulted in comparable cross-reacting antibodies to the antibodies induced both by vaccination in cardiac patients and by natural infection in military conscripts. After a steady decline, the cross-reactivity to the latest epidemic virus improved in all the groups. This may be due to two reverted amino acid changes in the HA1 domain of the virus haemagglutinin. CONCLUSIONS: Influenza vaccination in HD patients is as effective as the vaccination of cardiac patients with normal kidney function. The cross-reactivity of vaccination induced antibodies is even as good as that of antibodies induced by natural infection of young healthy males. Additionally, vaccination seems to prime the individual beneficially against subsequent years' influenza viruses. PMID- 12637649 TI - Effect of the duration of dialysis on survival in a cohort of chronic haemodialysis patients. AB - BACKGROUND: Atherosclerosis and vascular calcification are common in chronic haemodialysis (HD) patients, and usually progress with time. Whether the length of dialysis treatment in chronic HD patients is a significant independent risk factor of death is not clear. METHODS: A cohort of chronic HD patients from the Okinawa Dialysis Study, n=1243 (720 men, 523 women), was followed from January 1991 to December 2000, and their survival rates were compared against the duration of HD, which was calculated in months from the start of dialysis therapy to January 1991. A Cox proportional hazards regression analysis was done to examine the influence of the duration of dialysis on survival, after adjusting for other factors such as age, sex, serum albumin concentration and diastolic blood pressure. The hazards ratio and 95% confidence interval (CI) were calculated in both diabetic and non-diabetic patients. RESULTS: The mean duration of dialysis was 61.9 months and ranged from 1 to 233 months. The numbers of patients who died, underwent renal transplantation or were transferred outside Okinawa were 568 (45.7%), 61 (4.9%) and 14 (1.1%), respectively, during the study. The hazards ratio (95% CI) was 1.002 (1.000-1.004, P=0.0245) for non diabetic patients and 1.006 (1.001-1.011, P=0.0214) for diabetic patients, suggesting that the longer the duration of dialysis, the greater the risk of death. CONCLUSIONS: This study shows that prolonged dialysis is a significant predictor of death in chronic HD patients, in particular diabetic patients. Whether this is related to the progression of the atherosclerotic process or to uraemic conditions remains to be shown. PMID- 12637650 TI - Personal dialysis capacity (PDC(TM)) test: a multicentre clinical study. AB - BACKGROUND: The assessment of the peritoneal membrane capacity and physiology of the individual patient is becoming increasingly important. It allows the prescription of an individualized peritoneal dialysis (PD)-regimen, and the monitoring of peritoneal membrane function over time. The PDC(TM) program offers the possibility to evaluate the peritoneal membrane characteristics and to predict solute and water removal by simulation of different treatment regimens. METHODS: This study evaluates the relevance of the PDC(TM) program when routinely used. The PDC(TM) data of 336 patients from nine different centres in Europe were evaluated. RESULTS: The area parameter was 20 985+/-7578 cm/1.73 m(2) (mean+/ SD). The reabsorption of fluid after dissipation of glucose, Jv(AR), was 1.97+/ 1.00 ml/min/1.73 m(2). The large pore fluid flux, Jv(L), was 0.11+/-0.07 ml/min/1.73 m(2). A multivariate model for prediction of serum albumin included dialysate protein loss, Jv(L), Jv(AR), nPCR, A(0)/deltaX, BMI and gender (R(2)=0.81, P<0.001). Total clearance fell with increasing PD duration (P<0.001). A negative relation between A(0)/deltaX and ultrafiltration (rho=-0.26, P<0.05), a positive relation between A(0)/deltaX and peritoneal creatinine clearance (rho=0.52, P<0.05) and urea clearance (rho=0.36, P<0.05), and a positive relation between measured peritoneal creatinine and urea clearance (rho=0.64, P<0.01) was observed. CONCLUSIONS: In summary, the present study shows that the PDC(TM) program is a robust, accurate method to describe the peritoneal membrane transport characteristics. Analysis of PDC(TM) data of large groups of patients, especially if followed up over time, can give interesting information on the physiology of the peritoneal membrane and the impact of different parameters on it. PMID- 12637651 TI - Fluid status in CAPD patients is related to peritoneal transport and residual renal function: evidence from a longitudinal study. AB - BACKGROUND: Both peritoneal transport characteristics as well as residual renal function are related to outcome in patients treated with continuous ambulatory peritoneal dialysis (CAPD). It has been suggested that part of this relationship might be explained by an effect of both parameters on the fluid state in CAPD patients or by the relationship between inflammation and peritoneal transport. METHODS: In the present study, the relationship between fluid state [extracellular water (ECW) (sodium bromide); total body water (TBW) (deuterium oxide)] with peritoneal transport characteristics (2.27% glucose dialysate/plasma creatinine [D/P (creat)] ratio), residual renal function (residual glomerular filtration rate [rGFR] by urine collection) and C-reactive protein (CRP) was assessed in 37 CAPD patients in a cross-sectional and longitudinal design, with 25 patients completing the study. RESULTS: In the cross-sectional part ECW, corrected for height (ECW:height), was inversely related to rGFR (r=-0.40, P=0.016), whereas during the longitudinal part, D/P[creat] was related to the change in ECW (r=0.40, P=0.05). Neither D/P[creat] nor rGFR were related to CRP, whereas a significant relationship was observed between ECW:height and CRP (r=0.58, P=0.0001). Patients were dichotomized according to rGFR (<2 or >2 ml/min). Despite a higher daily peritoneal glucose prescription (216.3+/-60.0 vs 156.5+/-53.0 g/24 h; P=0.004) and peritoneal ultrafiltration volume (1856+/-644 vs 658+/-781 ml/24 h, respectively; P=0.0001), the patients with a rGFR <2 ml/min showed a higher ECW:height compared with the group with rGFR >2 ml/min (12.5+/ 3.8 vs 9.2+/-2.2 l/m, respectively; P=0.003). Results for TBW were comparable. CONCLUSION: Fluid state was significantly related to peritoneal transport characteristics and rGFR. The larger ECW:height in CAPD patients with a negligible rGFR existed despite a higher peritoneal ultrafiltration volume and higher peritoneal glucose prescription. These findings raise doubts as to whether fluid state in CAPD patients with a diminished rGFR can be adequately controlled on standard glucose solutions without an additional sodium and fluid restriction. The preliminary finding of a relationship between CRP and fluid state might suggest a relationship between overhydration and inflammation. PMID- 12637652 TI - Video-assisted thoracoscopic talc pleurodesis is effective for maintenance of peritoneal dialysis in acute hydrothorax complicating peritoneal dialysis. AB - BACKGROUND: Acute, massive, unilateral hydrothorax is an uncommon but well recognized complication of peritoneal dialysis. Its clinical course and treatment outcome after a recently advocated technique of video-assisted thoracoscopic (VATS) talc pleurodesis remains unclear. METHODS AND RESULTS: Between July 1998 and March 2002, among 475 CAPD patients in two regional hospitals in Hong Kong, nine patients (three men, six women, mean age 53+/-12 years) developed acute hydrothorax due to pleuroperitoneal communication (R=8, L=1) within 5.8+/-4.2 months (median, 5.2 m; range, 2 days to 11.6 months) of commencing peritoneal dialysis. Analysis of simultaneously obtained peritoneal and pleural fluid in all subjects only showed concordance in protein content (consistently<4 g/l), while fluid glucose and lactate dehydrogenase levels were not comparable. The methylene blue test was negative (n=4). Radionuclide scan (n=6) and contrast CT peritoneography (CTP, n=3) detected pleuroperitoneal communication in half and one-third of the patients, respectively. All patients underwent pleurodesis achieved by talc insufflation into the pleural cavity under VATS guidance. All patients were successfully returned to peritoneal dialysis. After a mean follow up of 18.8+/-12.5 months, hydrothorax recurred in one patient (at 7 months after pleurodesis), who was successfully treated by repeating the procedure. CONCLUSIONS: Hydrothorax complicating CAPD is more commonly right-sided, and tends to occur within the first year of starting peritoneal dialysis. Isotope scan and CTP are insensitive in diagnosing pleuroperitoneal communication. A low pleural fluid protein content is the most consistent biochemical finding. VATS talc pleurodesis is a safe and reliable treatment of choice that allows sustained continuation of CAPD with low recurrence rate. PMID- 12637653 TI - Comparative study of prophylactic oral ganciclovir and valacyclovir in high-risk kidney transplant recipients. AB - BACKGROUND: Cytomegalovirus (CMV) is a major pathogen in renal transplant patients causing significant post-transplant morbidity and mortality. Prophylactic antiviral therapy, currently implemented in most kidney transplant centres, has significantly reduced the incidence of CMV infection after transplantation. Oral ganciclovir has been shown to be an effective prophylactic agent in preventing CMV disease and infection with a demonstrated superior efficacy over oral acyclovir. Valacyclovir, a prodrug of acyclovir with a higher level of bioavailability than acyclovir, has also been shown to be effective in preventing CMV disease when given as prophylactic treatment. METHODS: In a retrospective analysis of 150 renal transplant recipients in our centre, we compared the efficacy of oral ganciclovir with valacyclovir in preventing CMV infection. Seventy-seven consecutive renal transplant recipients prophylactically treated with oral ganciclovir for 12 weeks after transplant were compared with 73 consecutive recipients treated with oral valacylovir for an equal length of time. RESULTS: No difference was noted in the incidence of CMV infection between the two treatment groups (5.1 vs 5.4%) after a 6 month follow-up. Likewise, the incidence of acute rejection was similar in both groups (11.6 vs 6.8%). All cases of CMV infection occurred in high-risk patients (donor positive/recipient negative). CONCLUSION: The prophylactic use of oral valacylovir is as effective as oral ganciclovir in reducing CMV infection and disease after kidney transplantation. PMID- 12637654 TI - Detection of cell-cycle regulators in failed arteriovenous fistulas for haemodialysis. AB - BACKGROUND: Chronic renal failure (CRF) patients rely on arteriovenous fistulas (AVFs) for haemodialysis vascular access. Intimal hyperplastic stenoses result in failure of AVFs and frequent intervention is required to maintain vascular access. The extent of intimal hyperplasia depends on the interplay between cyclins and cyclin-dependent kinases (e.g. cdk2), positively regulating cell cycle progression. cdk activity is negatively modulated by the interaction with cdk inhibitory proteins, such as p21(Waf1) and p27(Kip1). Little is known about the expression of these proteins in the development of intimal hyperplasia in AVFs. METHODS: p21(Waf1), p27(Kip1), cdk2 and Proliferating Cell Nuclear Antigen immunoreactivity was determined in 18 failed AVFs from 16 CRF patients and 10 non diseased vessels (five arteries and five veins). RESULTS: The percentage of p21(Waf1)-positive cells was significantly lower in AVFs (3+/-1%), compared with normal veins and arteries (62+/-4 and 63+/-4%, respectively; P<0.001). cdk2 positive cells were significantly higher in AVFs (40.7+/-3.7%) than in normal veins and arteries (2+/-1 and 0+/-0%, respectively; P<0.001). Although no difference in p27(Kip1) immunoreactivity was found between AVFs (37+/-17%) and veins (23+/-8%; P=0.208), it was lower in healthy arteries (17+/-11%; P=0.037). CONCLUSIONS: The data suggest that in failed AVFs, p21(Waf1), but not p27(Kip1), is related to intimal hyperplasia. This is the first report to show involvement of cell-cycle regulators in AVF-related human intimal hyperplasia. PMID- 12637656 TI - WT1 splice site mutation in a 46,XX female with minimal-change nephrotic syndrome and Wilms' tumour. PMID- 12637655 TI - Impact of St John's wort treatment on the pharmacokinetics of tacrolimus and mycophenolic acid in renal transplant patients. AB - BACKGROUND: This study investigated the effect of St John's wort (SJW) extract on the pharmacokinetics of the immunosuppressants tacrolimus (TAC) and mycophenolic acid (MPA). METHODS: Ten stable renal transplant patients received 600 mg SJW extract for 14 days in addition to their regular regimen of TAC and mycophenolate mofetil. RESULTS: Dose-corrected AUC((0-12)) of TAC decreased significantly from 180 ng/ml/h at baseline to 75.9 ng/ml/h after 2 weeks of SJW treatment. To maintain therapeutic TAC concentrations, dose adjustments from a median 4.5 mg/day at baseline to 8.0 mg/day under SJW treatment were required. Two weeks after discontinuation of SJW, TAC doses were reduced to a median of 6.5 mg/day. MPA pharmacokinetics remained unaffected by comedication with hypericum extract. CONCLUSIONS: Administration of SJW extract to patients receiving TAC treatment can result in a serious drug interaction leading to markedly reduced TAC blood concentrations associated with the risk of organ rejection. PMID- 12637657 TI - Pamidronate-induced tubulointerstitial nephritis with Fanconi syndrome in a patient with primary hyperparathyroidism. PMID- 12637658 TI - Use of an intra-aortic Tesio catheter as vascular access for haemodialysis. PMID- 12637659 TI - Acute renal failure caused by unilateral renal artery thromboembolism. PMID- 12637660 TI - A wretching business: 'how to get the most out of the numbers'. PMID- 12637661 TI - Acute renal failure in a patient suffering from chronic alcoholism. PMID- 12637662 TI - Myoclonic jerks in a post-transplant patient: is tacrolimus the sole culprit? PMID- 12637663 TI - Ultrasonographic fata morgana. PMID- 12637664 TI - A Christmas dialysis vignette. PMID- 12637666 TI - Management of hyperphosphataemia in chronic renal disease: lessons from the past and future directions. PMID- 12637667 TI - Comparison in renal histology between Japanese obese and non-obese microalbuminuric type 2 diabetic patients. PMID- 12637668 TI - Acute septic arthritis after kidney transplantation due to Acremonium. PMID- 12637669 TI - Kidney transplantation in patients travelling from the UK to India or Pakistan. PMID- 12637671 TI - Pyogenic bacterial infections in humans with IRAK-4 deficiency. AB - Members of the Toll-like receptor (TLR) and interleukin-1 receptor (IL-1R) superfamily share an intracytoplasmic Toll-IL-1 receptor (TIR) domain, which mediates recruitment of the interleukin-1 receptor-associated kinase (IRAK) complex via TIR-containing adapter molecules. We describe three unrelated children with inherited IRAK-4 deficiency. Their blood and fibroblast cells did not activate nuclear factor kappaB and mitogen-activated protein kinase (MAPK) and failed to induce downstream cytokines in response to any of the known ligands of TIR-bearing receptors. The otherwise healthy children developed infections caused by pyogenic bacteria. These findings suggest that, in humans, the TIR-IRAK signaling pathway is crucial for protective immunity against specific bacteria but is redundant against most other microorganisms. PMID- 12637672 TI - Ultrahigh-density nanowire lattices and circuits. AB - We describe a general method for producing ultrahigh-density arrays of aligned metal and semiconductor nanowires and nanowire circuits. The technique is based on translating thin film growth thickness control into planar wire arrays. Nanowires were fabricated with diameters and pitches (center-to-center distances) as small as 8 nanometers and 16 nanometers, respectively. The nanowires have high aspect ratios (up to 10(6)), and the process can be carried out multiple times to produce simple circuits of crossed nanowires with a nanowire junction density in excess of 10(11) per square centimeter. The nanowires can also be used in nanomechanical devices; a high-frequency nanomechanical resonator is demonstrated. PMID- 12637673 TI - The pentacovalent phosphorus intermediate of a phosphoryl transfer reaction. AB - Enzymes provide enormous rate enhancements, unmatched by any other type of catalyst. The stabilization of high-energy states along the reaction coordinate is the crux of the catalytic power of enzymes. We report the atomic-resolution structure of a high-energy reaction intermediate stabilized in the active site of an enzyme. Crystallization of phosphorylated beta-phosphoglucomutase in the presence of the Mg(II) cofactor and either of the substrates glucose 1-phosphate or glucose 6-phosphate produced crystals of the enzyme-Mg(II)-glucose 1,6 (bis)phosphate complex, which diffracted x-rays to 1.2 and 1.4 angstroms, respectively. The structure reveals a stabilized pentacovalent phosphorane formed in the phosphoryl transfer from the C(1)O of glucose 1,6-(bis)phosphate to the nucleophilic Asp8 carboxylate. PMID- 12637674 TI - Chemistry. Seeing is believing. PMID- 12637675 TI - Infantile hypertrophic pyloric stenosis. AB - Infantile hypertrophic pyloric stenosis is a common condition affecting young infants; despite its frequency, it has been recognized only for a little over a century, and its etiology remains unknown. Nevertheless, understanding of the condition and of effective treatment have undergone a remarkable evolution in the 20th century, reducing the mortality rate from over 50% to nearly 0%. The lesion is characterized by gastric outlet obstruction and multiple anatomic abnormalities of the pyloric antrum. The antropyloric muscle is abnormally thickened and innervated, and the intervening lumen is obstructed by crowded and redundant mucosa. Recognition of the obstructive role of the mucosa led to discovery of effective surgical treatment. Accurate clinical diagnosis in patients in whom a thickened antropyloric muscle is not readily palpable can be difficult, resulting in delayed diagnosis and can lead to emaciation and electrolyte imbalance, making the patient a suboptimal surgical candidate. Current imaging techniques, particularly sonography, are noninvasive and accurate for identification of infantile hypertrophic pyloric stenosis. Successful imaging requires understanding of anatomic changes that occur in patients with this condition and plays an integral role in patient care. Accurate, rapid, noninvasive imaging techniques facilitate rapid referral of vomiting infants and prompt surgical treatment of more suitable surgical candidates. PMID- 12637676 TI - Preoperative portal vein embolization with a new liquid embolic agent. AB - PURPOSE: To evaluate the effectiveness and safety of a new liquid embolic agent in preoperative portal vein embolization. MATERIALS AND METHODS: Embol-78 was obtained by means of hydrolysis of polyvinyl acetate and was dissolved in a mixture of ethanol and nonionic water-soluble contrast medium. After percutaneous puncture of the portal vein, embolization of the right portal vein was performed in 22 patients with hepatocellular carcinoma and in 29 patients with nonhepatocellular carcinoma. In each group, changes in volume of the future liver remnant, portal venous pressure, and liver enzymes were evaluated both before and after embolization. Complications were also evaluated. RESULTS: Portal vein embolization was successful in all patients, without major complications. The mean volumes of the future liver remnant before and 2 weeks after embolization were 385 mL +/- 138 and 533 mL +/- 140, respectively, in the hepatocellular carcinoma group and 517 mL +/- 348 and 755 mL +/- 197, respectively, in the nonhepatocellular carcinoma group. There were only transient elevations in liver enzyme levels after embolization. Mean portal venous pressures before and after the procedure were 16.7 mm Hg +/- 3.8 and 20.3 mm Hg +/- 3.6, respectively, in the hepatocellular carcinoma group and 11.7 mm Hg +/- 3.5 and 14.6 mm Hg +/- 3.6, respectively, in the nonhepatocellular carcinoma group. In each group, changes in volume of the future liver remnant and portal venous pressure were statistically significant (P <.001). CONCLUSION: The liquid embolic material Embol-78 seems to be effective and safe for preoperative portal vein embolization. PMID- 12637677 TI - Neurologic complications of cerebral angiography: prospective analysis of 2,899 procedures and review of the literature. AB - PURPOSE: To prospectively identify risk factors for neurologic complications related to cerebral angiography. MATERIALS AND METHODS: A total of 2,899 consecutive cerebral digital subtraction angiograms obtained with nonionic contrast material were prospectively evaluated. Neurologic complications were categorized as transient (<24 hours), reversible (24 hours to 7 days), and permanent (>7 days). The neurologic complication rate was correlated with patient age, type of indication for catheter angiography, medical history, fluoroscopic time, number and size of catheters, type and number of vessels injected, operator experience, and the quartile in which the study was performed. The correlations were statistically analyzed with Fisher exact tests and a multiple logistic regression model. RESULTS: There were 39 (1.3%) neurologic complications in 2,899 procedures; 20 were transient (0.7%), five (0.2%) were reversible, and 14 (0.5%) were permanent. Neurologic complications were significantly more common in patients 55 years of age or older (25 of 1,361; 1.8%) (P =.035), in patients with cardiovascular disease (CVD) (20 of 862; 2.3%) (P =.004), and when fluoroscopic times were 10 minutes or longer (24 of 1,238; 1.9%) (P =.022). The neurologic complication rate was higher in procedures performed by fellows alone (24 of 1,878; 1.3%) compared with that when staff alone performed the procedures (three of 598; 0.5%), but the difference was not significant (P =.172). Neurologic complications were lower in the fourth quartile of the study (six of 171; 0.9%) compared with the first quartile (16 of 776; 2.1%), which was likely due to fewer patients being examined for carotid stenosis or ischemic stroke and fewer patients with CVD (P =.085). CONCLUSION: Age-related vascular disease accounted for the failure to lower the neurologic complication rate of cerebral angiography despite technical advances. PMID- 12637678 TI - Thoracic outlet: assessment with MR imaging in asymptomatic and symptomatic populations. AB - PURPOSE: To compare the dynamic modifications of the thoracic outlet in asymptomatic volunteers and symptomatic patients and assess the presence and location of vasculonervous compressions in these two populations. MATERIALS AND METHODS: Thirty-five healthy volunteers and 54 patients with clinical symptoms of thoracic outlet syndrome (TOS) underwent magnetic resonance (MR) imaging of the thoracic outlets with their arms alongside their bodies and after a postural maneuver. Measurements were obtained at the interscalene triangle (thickness of anterior scalene muscle, interscalene angle), at the costoclavicular space (minimum costoclavicular distance, distance between inferior border of subclavius muscle and the anterior chest wall, maximum thickness of subclavius muscle, angle between first rib shaft and horizontal), and at the retropectoralis minor space (distance between posterior border of pectoralis minor muscle and posterior lining of axilla at the passage of the axillary vessels, thickness of pectoralis minor muscle). The presence and location of vasculonervous compressions were also assessed. Group data were analyzed with the Student t test. RESULTS: Patients with TOS had a smaller costoclavicular distance after the postural maneuver (P <.001), a thicker subclavius muscle in both arm positions (P <.001), and a wider retropectoralis minor space after the postural maneuver (P <.001) than did volunteers. Venous compressions after the postural maneuver were observed in 47% of volunteers and 63% of patients at the prescalene space, in 54% of volunteers and 61% of patients at the costoclavicular space, and in 27% of volunteers and 30% of patients at the retropectoralis minor space. Arterial and nervous compressions, respectively, were seen in 72% and 7% of patients. No arterial or nervous compression was seen in volunteers. Except for venous thrombosis, vasculonervous compressions were demonstrated only with arm elevation. Only three thoracic outlet measurements differed significantly in both populations. CONCLUSION: MR imaging appeared helpful in demonstrating the location and cause of arterial or nervous compressions. PMID- 12637679 TI - Camera-based FDG PET and 67Ga SPECT in evaluation of lymphoma: comparative study. AB - PURPOSE: To compare gallium 67 (67Ga) scintigraphy and camera-based fluorodeoxyglucose (FDG) positron emission tomography (PET) in the evaluation of patients with lymphoma. MATERIALS AND METHODS: The performance of 67Ga scintigraphy and camera-based FDG PET in the detection of lymphoma was retrospectively evaluated and compared in 84 patients with lymphoma, with 219 suspected sites of disease. Eighty-nine percent of patients were examined during or after treatment. Camera-based FDG PET was initiated by equivocal characterization of the status of disease based on clinical, radiologic, and 67Ga scintigraphic assessment. Findings of 67Ga scintigraphy and camera-based FDG PET were compared on a per-patient and per-site basis for the whole group, for histologic subtypes, and for anatomic locations. Comparison of sensitivity, specificity, and accuracy between the two modalities for detection of lymphoma was performed with the McNemar test. RESULTS: There was a statistically significant difference in sensitivity, specificity, and accuracy of 67Ga scintigraphy and camera-based FDG PET at both patient- and site-based analysis. 67Ga scintigraphy helped to accurately define disease state in 63% of patients and in 33% of sites, compared with 83% and 87%, respectively, for camera-based FDG PET. For discordant findings between the two modalities, camera-based FDG PET findings were confirmed as true-positive in 71% and as true-negative in 92% of patients. Camera-based FDG PET had a significantly higher detection rate for both nodal and extranodal lymphoma sites. It provided accurate assessment of lymphoma involvement of the skeleton in 93% of sites compared with 29% for 67Ga scintigraphy and excluded active lymphoma in 10 67Ga-positive benign parahilar sites. CONCLUSION: In this selected group of patients with lymphoma, camera-based FDG PET allowed a significantly more accurate definition of active disease compared with that allowed with 67Ga scintigraphy. PMID- 12637680 TI - Acute cholecystitis at T2-weighted and manganese-enhanced T1-weighted MR cholangiography: preliminary study. AB - Twelve patients with symptoms of acute cholecystitis underwent heavily T2 weighted and mangafodipir trisodium-enhanced T1-weighted magnetic resonance (MR) cholangiography and cholescintigraphy before they underwent cholecystectomy. On T2-weighted MR cholangiographic images, morphologic evidence of outflow obstruction of the gallbladder was definitive in seven patients, equivocal in one, and absent in four. In all 12 patients, biliary dynamics depicted at manganese-enhanced T1-weighted MR cholangiography agreed completely with those depicted at hepatobiliary scintigraphy. T2-weighted MR cholangiography combined with manganese-enhanced T1-weighted MR cholangiography provides not only morphologic information but also functional information about the biliary system. PMID- 12637681 TI - Mammalian basal metabolic rate is proportional to body mass2/3. AB - The relationship between mammalian basal metabolic rate (BMR, ml of O(2) per h) and body mass (M, g) has been the subject of regular investigation for over a century. Typically, the relationship is expressed as an allometric equation of the form BMR = aM(b). The scaling exponent (b) is a point of contention throughout this body of literature, within which arguments for and against geometric (b = 2/3) and quarter-power (b = 3/4) scaling are made and rebutted. Recently, interest in the topic has been revived by published explanations for quarter-power scaling based on fractal nutrient supply networks and four dimensional biology. Here, a new analysis of the allometry of mammalian BMR that accounts for variation associated with body temperature, digestive state, and phylogeny finds no support for a metabolic scaling exponent of 3/4. Data encompassing five orders of magnitude variation in M and featuring 619 species from 19 mammalian orders show that BMR proportional, variant M(2/3). PMID- 12637682 TI - An engineered two-iron superoxide reductase lacking the [Fe(SCys)4] site retains its catalytic properties in vitro and in vivo. AB - Superoxide reductases (SORs) contain a characteristic square-pyramidal [Fe(NHis)(4)(SCys)] active site that catalyzes reduction of superoxide to hydrogen peroxide in several anaerobic bacteria and archaea. Some SORs, referred to as two-iron SORs (2Fe-SORs), also contain a lower-potential [Fe(SCys)(4)] site that is presumed to have an electron transfer function. However, the intra- and inter-subunit distances between [Fe(SCys)(4)] and [Fe(NHis)(4)(SCys)] iron centers within the 2Fe-SOR homodimer seem too long for efficient electron transfer between these sites. The possible role of the [Fe(SCys)(4)] site in 2Fe SORs was addressed in this work by examination of an engineered Desulfovibrio vulgaris 2Fe-SOR variant, C13S, in which one ligand residue of the [Fe(SCys)(4)] site, cysteine 13, was changed to serine. This single amino acid residue change destroyed the native [Fe(SCys)(4)] site with complete loss of its iron, but left the [Fe(NHis)(4)(SCys)] site and the protein homodimer intact. The spectroscopic, redox and superoxide reactivity properties of the [Fe(NHis)(4)(SCys)] site in the C13S variant were nearly indistinguishable from those of the wild-type 2Fe-SOR. Aerobic growth complementation of a superoxide dismutase (SOD)-deficient Escherichia coli strain showed that the presence of the [Fe(NHis)(4)(SCys)] site in C13S 2Fe-SOR was apparently sufficient to catalyze reduction of the intracellular superoxide to nonlethal levels. As is the case for the wild-type protein, C13S 2Fe-SOR did not show any detectable SOD activity, i.e., destruction of the [Fe(SCys)(4)] site did not unmask latent SOD activity of the [Fe(NHis)(4)(SCys)] site. Possible alternative roles for the [Fe(SCys)(4)] site in 2Fe-SORs are considered. PMID- 12637684 TI - Finding the best ways to help: opportunities and challenges of intervention research on aging. PMID- 12637683 TI - Yeast genetic selections to optimize RNA decoys for transcription factor NF-kappa B. AB - In vitro-selected RNA aptamers are potential inhibitors of disease-related proteins. Our laboratory previously isolated an RNA aptamer that binds with high affinity to human transcription factor NF-kappaB. This RNA aptamer competitively inhibits DNA binding by NF-kappaB in vitro and is recognized by its target protein in vivo in a yeast three-hybrid system. In the present study, yeast genetic selections were used to optimize the RNA aptamer for binding to NF-kappaB in the eukaryotic nucleus. Selection for improved binding to NF-kappaB from RNA libraries encoding (i) degenerate aptamer variants and (ii) sequences present at round 8 of 14 total rounds of in vitro selection yielded RNA aptamers with dramatically improved in vivo activity. Furthermore, we show that an in vivo optimized RNA aptamer exhibits specific "decoy" activity, inhibiting transcriptional activation by its NF-kappaB target protein in a yeast one-hybrid assay. This decoy activity is enhanced by the expression of a bivalent aptamer. The combination of in vitro and in vivo genetic selections was crucial for obtaining RNA aptamers with in vivo decoy activity. PMID- 12637685 TI - Practically relevant research: capturing real world tasks, environments, and outcomes. AB - PURPOSE: Development of strategies to optimize the functional performance of older adults requires understanding the behavior of older people doing tasks in real-world settings and capturing these interactions in research protocols. This is a major challenge as there is some degree of tension between capturing the contextual variables and constraints that operate in the real world and the scale of research that can be realistically conducted within controlled experimental settings. This article presents a research approach that can be used to help ensure the ecological validity of research protocols. The intent is to demonstrate how an ecologically valid approach affords greater insight into the performance of older adults in real world settings. METHODS: The approach involves techniques such as task analysis and simulation. Examples from two research projects examining aging and the performance of real-world computer based work tasks are used to demonstrate the application of this approach. RESULTS: The article demonstrates how an ecologically valid research approach yields information about human performance that can be translated into solutions for real-world problems. IMPLICATIONS: Ecologically valid research protocols offer the potential for providing answers to real-world problems and advancing theory regarding aging and functional performance. PMID- 12637686 TI - Integrating theory, basic research, and intervention: two case studies from caregiving research. AB - PURPOSE: The premise of this article is that interventions should be based explicitly on theory and basic research findings. Although there appears to be general agreement with that assertion, the connections among theory, research, and intervention in the field of gerontology are often tenuous or nonexistent. In this article, we argue for better integration of these three domains, providing two case studies from the Cornell Roybal Center that illustrate the positive role theory and research can play in intervention designs and broader applicability of findings. DESIGN AND METHODS: Study 1 involved a social support intervention for persons making the transition to becoming a family caregiver. Study 2 was an organizational intervention designed to improve interpersonal relationships and increase mutual support between family caregivers and staff in nursing homes. RESULTS: Several benefits emerged as a result of creating theoretically grounded and research-based interventions, including guidance for innovative intervention design and the production of findings that inform both basic research and intervention. IMPLICATIONS: A much closer link between theory and basic research and intervention studies is indicated, suggesting that current federal support of translational research initiatives is justified and worthy of expansion. PMID- 12637687 TI - Obstacles to implementing research outcomes in community settings. AB - PURPOSE: In contrast to controlled laboratory- or clinic-based research that can fail to capture the real-world behaviors of older adults, field research offers the best opportunity for ecological validity. However, the tradeoff inherent in field studies is the potential sacrifice of scientific rigor. Applied research presents a unique set of challenges that vary with context. This article discusses these challenges along with possible solutions. DESIGN: Examples are drawn from an ongoing, longitudinal Roybal Center study of driving competence that is being conducted in Department of Motor Vehicles field sites. The challenges faced at each stage of the project are discussed. METHOD: Methodological issues include identifying field collaborators, approaching administrators with the research proposal, producing a battery that is manageable and acceptable while maintaining scientific merit, training indigenous personnel to administer this battery, introducing the research and consenting potential participants, and managing large data sets offsite. Additional issues include quality control, the importance of distinguishing between individuals who consent and those who decline participation, and the collection of follow-up data via telephone. IMPLICATIONS: The use of field research in changing public perceptions, medical practice, and public policy is discussed. PMID- 12637689 TI - Knowledge dissemination and utilization in gerontology: an organizing framework. AB - PURPOSE: Enabling valuable research findings to be used by the field requires a strategic approach to dissemination and utilization rather than simply making constituencies aware of the information. This article describes a conceptual framework for the dissemination and utilization of information, along with examples of its use by the Boston University Roybal Center for Enhancement of Late Life Function. DESIGN AND METHODS: The framework identifies dissemination/utilization goals of exposure, experience, expertise, and embedding ("4 E") and relates each goal to strategies targeted for specific users. RESULTS AND IMPLICATIONS: The Boston University center exposed the field to information through presentations, print- and Web-based information, provided consumers and family members with new findings through motivational videotapes, developed expertise-level training programs, and embedded the new findings within organizations and systems. The 4 E framework can translate critical research outcomes into useful information to assist the field to better care and support available for individuals in late life. PMID- 12637688 TI - Challenges and opportunities in recruiting and retaining underrepresented populations into health promotion research. AB - PURPOSE: To evaluate how recruitment strategies and program characteristics interact with participant characteristics to influence recruitment and retention in an exercise intervention study targeted to African American and White older adults with multiple chronic illnesses. DESIGN AND METHODS: Characteristics of 273 referrals and 103 enrollees were analyzed in conjunction with programmatic decisions about recruitment design and eligibility criteria. RESULTS: Eligible participants who did not enroll were younger and more likely to be under 60 and to self-report having diabetes. After 1 year, 70% of the enrolled participants remained in the program. Program attrition was not associated with randomization, race, or chronic illness but was associated with functional status, having a high school degree, and program site. IMPLICATIONS: Program design decisions can significantly influence the participation of underrepresented populations in exercise health promotion programs for older adults. In particular, group specific efforts targeted to recruiting and retaining African Americans can be successful. PMID- 12637690 TI - Theoretical models of cognitive aging and implications for translational research in medicine. AB - PURPOSE: We provide an overview of theoretical models of cognitive aging and present empirical research that uses these models to explain older patients' medical behaviors and to develop interventions for improving the delivery of health information and services to older adults. DESIGN AND METHODS: Theoretical accounts of age and cognition are summarized and are related to key research findings, including age differences in comprehension of medical information, adherence, and use of medical technologies. The implications of cognitive aging theories for designing optimal medical environments and enhancing adherence are discussed. RESULTS: Age declines in basic cognitive abilities such as working memory capacity limit older adults' ability to comprehend and recall several types of novel medical information. In contrast, automatic processes and environmental cues can benefit older adults, as shown by age increases in compliance for practiced or mentally imaged health behaviors, but can also be dangerous, as shown by older adults' greater belief in false but familiar health statements. Last, cognitive aging is shown to be a useful model for investigating cognitive disorders. IMPLICATIONS: Cognitive aging theories are important for understanding older patient behaviors and suggest age-related declines as well as gains in performance in medical domains. In addition, models of cognitive aging are useful in guiding the development of interventions that improve health care in older patients and in furthering our understanding of cognitive dysfunction in populations other than older adults. PMID- 12637691 TI - Fibrinogen gene promoter -455 A allele as a risk factor for lacunar stroke. AB - BACKGROUND AND PURPOSE: Elevated fibrinogen levels are suggested to increase the risk of myocardial infarction and stroke. Carriers of the A allele of the fibrinogen -455G/A polymorphism have increased plasma fibrinogen levels. We studied the association of this polymorphism with stroke subtype in the Stroke Aging Memory (SAM) cohort. METHODS: The SAM cohort comprises 486 consecutive patients 55 to 85 years of age who, 3 months after ischemic stroke, completed a detailed stroke assessment. Stroke subtypes were examined with MRI. -455G/A genotype was determined by polymerase chain reaction. MRI and genotype data were available for the 299 patients who constitute the present study population. RESULTS: Genotype distributions were 64.9% (GG), 31.8% (GA), and 3.3% (AA). In a logistic regression model with age, sex, hypertension, diabetes, hypercholesterolemia, hypertriglyceridemia, myocardial infarction, arrhythmia, atrial fibrillation, peripheral arterial disease, and smoking as possible confounders, there was a significant association between A+ genotype and >or=3 lacunar infarcts (odds ratio [OR], 2.57; 95% CI, 1.23 to 5.36; P=0.01). Hypertensive patients carrying the A allele had increased risk (OR, 4.24; 95% CI, 1.29 to 13.99; P=0.02) for >or=3 lacunar infarcts. A similar increase in risk was observed among smokers with the A+ genotype (OR, 2.67; 95% CI, 0.92 to 7.77; P=0.07). CONCLUSIONS: Stroke patients carrying the A allele of the Bbeta fibrinogen -455G/A polymorphism frequently presented with multiple lacunar infarcts. This association was stronger among hypertensives and smokers. These associations suggest that the A allele may predispose to atherothrombotic events in cerebrovascular circulation. PMID- 12637692 TI - High-density lipoprotein cholesterol and risk of stroke in Japanese men and women: the Oyabe Study. AB - BACKGROUND AND PURPOSE: Evidence of an inverse relationship between serum high density lipoprotein cholesterol (HDL-C) and the risk of stroke is sparse in Asians and in women. The purpose of this investigation was to examine the relationship in a long-term cohort study of Japanese men and women among whom stroke occurrence is higher than in Western countries. METHODS: A prospective cohort study was performed involving 4989 participants (1523 men, 3466 women) 35 to 79 years of age at baseline with approximately 10 years of follow-up in a rural area of Japan. End points included all stroke incidence and ischemic stroke incidence. RESULTS: During follow-up, 132 participants developed stroke, including 81 ischemic stroke cases. Age-adjusted incidence rates per 10,000 person-years for all stroke in subjects with low HDL-C (<30 mg/dL [0.78 mmol/L]) were 103.4 in men and 49.3 in women, which were remarkably higher than in subjects with high HDL-C (>or=60 mg/dL [1.56 mmol/L]) (26.4 in men and 15.5 in women). A similar relationship was observed for ischemic stroke. Multivariate adjusted relative risks for all stroke incidence and ischemic stroke incidence were 2.89 (95% CI, 1.35 to 6.20) and 2.92 (95% CI, 1.17 to 7.32), respectively, for low versus high HDL-C participants. The relationships were independent of sex, age, body mass index, blood pressure, serum total cholesterol, alcohol consumption, and smoking. CONCLUSIONS: This 10-year follow-up study of Japanese men and women demonstrated that lower HDL-C levels were related significantly and independently to increased risk of all stroke incidence and ischemic stroke incidence. PMID- 12637693 TI - Spontaneous recanalization of internal carotid artery occlusion. AB - BACKGROUND AND PURPOSE: Spontaneous recanalization of an acutely occluded internal carotid artery (ICA) is an important phenomenon, the natural history and incidence of which have been incompletely studied. Although conventional catheter arteriography remains the gold standard for distinguishing total arterial occlusion from hairline residual lumen, CT angiography (CTA) is able to make this distinction noninvasively and more sensitively than either unenhanced MR angiography or ultrasound. The purpose of this report is to raise awareness of spontaneous recanalization and to demonstrate the possible use of CTA in following up cases of ICA occlusion. METHODS: We describe here 2 cases of acute cervical ICA occlusion seen on CTAs done at our institution. RESULTS: Follow-up CTAs in both cases showed spontaneous recanalization of the ICA requiring ipsilateral carotid endarterectomy within 1 month of the initial presentation. CONCLUSIONS: CTA, an accurate, rapid, and less invasive modality than conventional catheter arteriography, can be used to serially monitor anticoagulated patients with new-onset ICA occlusion for potential spontaneous vascular recanalization. The ability to conveniently assess ICA patency not only may influence management of individual patients but also could help us better establish the true incidence of spontaneous carotid recanalization in future studies. PMID- 12637694 TI - Internal carotid artery angle of origin: a novel risk factor for early carotid atherosclerosis. AB - BACKGROUND AND PURPOSE: Established "systemic" vascular risk factors do not fully explain the occurrence of atherosclerosis at the carotid bifurcation. Local anatomic and hemodynamic factors may also influence the initiation of the atherosclerotic process. We determined whether the angle of internal carotid artery (ICA) origin is a risk factor for early atherosclerosis. METHODS: In 1300 individuals from a normal population aged 40 to 70 years, we measured both carotid intima-media thickness (IMT) at 3 arterial sites (common carotid artery; carotid bifurcation; ICA bulb) and the presence of any atherosclerotic plaque within the ICA bulb bilaterally by means of high-resolution ultrasound. A standardized transverse insonation was used to determine the angle of ICA origin, expressed as the angle of rotation relative to the external carotid artery. RESULTS: This angle was positively associated with ICA bulb IMT but not with IMT at other sites. After we controlled for age, sex, and other cardiovascular risk factors, a dorsal/dorsomedial ICA origin (angle >or=60 degrees ) conferred an odds ratio for having an ICA bulb IMT in the highest quartile of 2.99 (95% CI, 1.86 to 4.83) on the left and 2.01 (95% CI, 1.31 to 3.09) on the right side (both P<0.001). A similar relationship was found for plaque; odds ratios on multivariate analysis were 3.67 (95% CI, 1.49 to 9.03) on the left and 2.07 (95% CI, 1.10 to 4.83) on the right side (both P=0.035). CONCLUSIONS: This study suggests that the angle of ICA origin may be an independent risk factor for early atherosclerotic changes at the ICA bulb. This new hypothesis should be tested in prospective studies. PMID- 12637695 TI - Detection of diffusion-weighted MRI abnormalities in patients with transient ischemic attack: correlation with clinical characteristics. AB - BACKGROUND AND PURPOSE: Although diffusion-weighted MRI (DWI) has demonstrated clear superiority over other conventional imaging modalities in the detection of hyperacute cerebral ischemia, its value in the evaluation of patients with transient symptoms has received only limited attention. We assessed the utility of DWI in patients with transient ischemic attack (TIA) to further evaluate the usefulness of this technique in these individuals. METHODS: A retrospective analysis was performed on all patients entered in the Stanford Stroke Center database during 1997-2001 who were clinically diagnosed with a TIA and who had also undergone a DWI scan or=1 hour, 16 times more likely to have had motor deficits, and 25 times more likely to have had aphasia than patients with negative DWI scans. The combination of all 3 symptoms was 100% specific for an abnormality on DWI. In 7 of 16 cases (44%), a DWI abnormality was present on both DWI and conventional imaging (T2-weighted imaging or fluid attenuated inversion recovery [FLAIR]). In all of these cases the DWI clarified the extent or acuity of the lesion (n=7) or identified additional lesions not detected by conventional imaging (n=9). CONCLUSIONS: In TIA patients, symptom duration >or=1 hour, motor deficits, and aphasia were each independently correlated with detecting an abnormality with DWI. DWI was also helpful in differentiating between chronic versus acute lesions. These data may be of value in identifying those TIA patients for whom MRI evaluation with DWI is of greatest clinical utility. PMID- 12637696 TI - Self-reported stress and risk of stroke: the Copenhagen City Heart Study. AB - BACKGROUND AND PURPOSE: Lay people often mention stress as one of the most important risk factors for stroke. Stress might trigger a cerebrovascular event directly or could be associated with higher levels of blood pressure or an unfavorable lifestyle. To examine these possibilities, we analyzed the association between self-reported stress frequency and intensity and risk of stroke. METHODS: Data from the second examination, 1981 to 1983, of participants in the Copenhagen City Heart Study were analyzed with Cox regression after a mean of 13 years of follow-up. A total of 5604 men and 6970 women were included, and 929 first-ever strokes occurred, of which 207 (22%) were fatal within 28 days after onset of symptoms. The stress frequency categories were never/hardly ever, monthly, weekly, or daily. The stress intensity categories were never/hardly ever, light, moderate, or high. RESULTS: Subjects with high stress intensity had almost a doubled risk of fatal stroke compared with subjects who were not stressed (relative risk [RR], 1.89; 95% CI, 1.11 to 3.21). Weekly stress was associated with an RR of 1.49 (95% CI, 1.00 to 2.23). There was no significant effect of stress in analyses of nonfatal strokes. Subjects who reported to be stressed often were more likely to have an adverse risk factor profile. CONCLUSIONS: Self-reported high stress intensity and weekly stress were associated with a higher risk of fatal stroke compared with no stress. However, there were no significant trends, and the present data do not provide strong evidence that self-reported stress is an independent risk factor for stroke. PMID- 12637697 TI - Synergistic effect of -174 G/C polymorphism of the interleukin-6 gene promoter and 469 E/K polymorphism of the intercellular adhesion molecule-1 gene in Italian patients with history of ischemic stroke. AB - BACKGROUND AND PURPOSE: Interleukin-6 (IL-6) and intercellular adhesion molecule 1 (ICAM-1) are involved in the pathogenetic mechanisms responsible for several ischemic cardiovascular disorders, including cerebral ischemia. IL-6 and ICAM-1 plasma levels and/or function may be genetically influenced. We sought to evaluate distribution and reciprocal interaction of IL-6 G/C gene promoter polymorphism and ICAM-1 E/K gene polymorphism in Italian patients with history of ischemic stroke. METHODS: One hundred nineteen patients with history of ischemic stroke and 133 age- and sex-matched controls were studied. IL-6 and ICAM-1 genotypes were evaluated by polymerase chain reaction and restriction enzyme analysis. RESULTS: The GG genotype of -174 IL-6 G/C gene polymorphism was significantly associated with history of ischemic stroke at both univariate (P<0.0001) and multivariate analysis (odds ratio [OR], 8.6; P<0.0001). Additionally, the EE genotype of ICAM-1 E/K gene polymorphism was significantly more common in the group of patients with history of ischemic stroke (P=0.003) and was an independent variable associated with stroke history (OR, 4.0; P=0.002). Interestingly, a further increased risk of stroke was found in subjects who concomitantly carry the IL-6 GG and ICAM-1 EE genotypes (IL-6 GG/ICAM-1 EE double-homozygous subjects) (OR, 10.1; P=0.004). CONCLUSIONS: There is a synergistic effect of IL-6 G/C and ICAM-1 E/K gene polymorphisms in patients with stroke history. Reciprocal interactions between genotypes may contribute in determining the risk profile for cardiovascular diseases and may merit further investigation as potential therapeutic targets. PMID- 12637698 TI - Secondary prevention of stroke in Italy: a cross-sectional survey in family practice. AB - BACKGROUND: Hypertension control and antiplatelet or oral anticoagulant drugs are the basis for secondary prevention of cerebrovascular events. Family physicians (FPs) are usually involved in both aspects of prevention, but no research has been carried out in Italy to evaluate the behavior of FPs in this field of prevention. METHODS: Data concerning 318 Italian FPs and 465,061 patients were extracted from the Health Search Database. Patients with coded diagnoses of stroke and transient ischemic attack (TIA) were selected. Demographic records and information regarding presence of concurrent disease and medical records were also obtained. Logistic regression analyses were carried out to assess whether conditions exist that make appropriate control of blood pressure (BP) and prescription of antiplatelet or anticoagulant drugs more likely. RESULTS: We selected 2555 patients with diagnosis of stroke and 2755 with TIA. Among all of the subjects, 32.6% had no BP recorded. Among the remaining subjects, 58.7% reported uncontrolled BP. Isolated systolic hypertension has been shown in 68.8% of patients with uncontrolled BP. Antiplatelet and anticoagulant drugs were prescribed in 72% of these cases. Factors that made the prescription significantly more unlikely were diagnosis of TIA (odds ratio [OR], 0.47; 95% confidence interval [CI], 0.41 to 0.54), total invalidity (OR, 0.66; 95% CI 0.56 to 0.78), and time from event of 5 years or more (OR, 0.81; 95% CI, 0.70 to 0.94). CONCLUSIONS: Italian FPs could improve secondary prevention of cerebrovascular accidents. The primary target of intervention should be the control of systolic BP, and the group of patients with unacceptably high BP should be given priority. All of these patients should have been prescribed antiplatelet drugs or anticoagulant agents, except in cases of extremely short life expectancy or substantial contraindications. PMID- 12637699 TI - Apolipoprotein E gene polymorphisms are associated with carotid plaque formation but not with intima-media wall thickening: results from the Perth Carotid Ultrasound Disease Assessment Study (CUDAS). AB - BACKGROUND AND PURPOSE: Several studies have investigated the role of apolipoprotein E (apoE) polymorphisms on carotid intima-media thickness (IMT) with conflicting results. The objective of this study was to use a large, community-based population to investigate associations between apoE gene polymorphisms and cardiovascular disease-associated phenotypes: IMT, carotid artery plaque, and low- (LDL-C) and high-density lipoprotein cholesterol (HDL-C). METHODS: ApoE genotypes were determined in 1109 randomly selected community subjects with an equal man-to-woman ratio and equal numbers in each age decile who were 27 to 77 years of age and had bilateral carotid B-mode ultrasound and cardiovascular risk factor measurements. RESULTS: Multivariate analyses, stratified by sex, demonstrated an association between apoE genotypes and LDL-C levels in men (P=0.03) and women (P<0.001). A significant linear trend in increasing LDL-C (beta=0.33 per unit change in genotype; SE=0.07; P<0.001) levels with increasing number of epsilon4 alleles across the epsilon3/epsilon3, epsilon3/epsilon4, or epsilon4/epsilon4 genotypes was observed in women but not in men. The associations were independent of age, diastolic blood pressure, and history of diabetes mellitus. Multivariate analyses found a log-additive trend in risk of developing carotid plaque with increasing numbers of epsilon4 alleles across the epsilon3/epsilon3, epsilon3/epsilon4, and epsilon4/epsilon4 genotypes (odds ratio [OR], 1.72 per unit change in genotype; 95% CI, 1.05 to 2.80; P=0.03) in men. There was no association between plaque frequency and the epsilon4 allele in women. However, the epsilon2/epsilon3 genotype was shown to be associated with a lower OR (OR, 0.40; 95% CI, 0.17 to 0.91; P=0.03) for carotid plaques relative to the epsilon3/epsilon3 genotype in women. The associations were independent of age and standard vascular risk factors. There were no significant independent associations between apoE genotypes and IMT in either men or women. CONCLUSIONS: Our data suggest that polymorphisms in the apoE gene are significantly associated with LDL-C levels and increased risk of carotid plaque formation in men but not IMT in either men or women. PMID- 12637700 TI - Nosocomial pneumonia after acute stroke: implications for neurological intensive care medicine. AB - BACKGROUND AND PURPOSE: Pneumonia has been estimated to occur in about one third of patients after acute stroke. Only limited data are available on stroke associated pneumonia (SAP) in specialized neurological intensive care units (NICUs). METHODS: We enrolled 124 patients with acute stroke who were treated at our university hospital NICU in a prospective observational study. Incidence rates and risk factors of SAP and long-term clinical outcome were determined. RESULTS: SAP incidence was 21% with a spectrum of pathogens, which is comparable to previously published data on general ICU patients. Mechanical ventilation, multiple location, and vertebrobasilar stroke, as well as dysphagia and abnormal chest x-ray findings, were identified as risk factors for the disease. SAP patients showed higher mortality rates than nondiseased subjects (acute, 26.9% versus 8.2%; long-term, 35.3% versus 14.3%) and a significantly poorer long-term clinical outcome (Barthel Index, 50.5+/-42.4 versus 81.5+/-27.8; Rankin Scale, 3.5+/-1.7 versus 2.2+/-1.6). CONCLUSIONS: Our data underline the considerable epidemiological and prognostic impact of SAP for the treatment of acute stroke patients in a specialized NICU setting. They demonstrate that the occurrence of SAP deteriorates clinical outcome in these patients. Our results allow us to identify high-risk stroke patients at time of NICU admission in whom the use of preventive treatment strategies is most promising. PMID- 12637701 TI - Epidemiological impact in the United States of a tissue-based definition of transient ischemic attack. AB - BACKGROUND AND PURPOSE: The traditional definition of transient ischemic attack (TIA), based on an arbitrary time criterion of symptom resolution within 24 hours, is problematic because a large number of patients with traditionally defined TIAs have a relevant cerebral infarction on brain imaging. The objective of this study was to characterize the epidemiological impact of adopting a tissue based definition of TIA. METHODS: Estimates of the annual US incidence of traditionally defined transient ischemic attacks were abstracted from the literature. Models were then constructed for determining the frequency of brain injury in traditionally defined TIAs, derived from recent human studies of MR diffusion-weighted imaging (DWI) in transient cerebral ischemia. RESULTS: Traditionally defined US TIA annual incidence rates ranged from 37 to 107 per 100,000 per year. Across 5 series, the raw frequency of DWI positivity in traditionally defined TIAs was 44%. Adjusting for an overrepresentation of longer duration TIAs in MR series yielded an expected frequency of diffusion MRI positivity of 33% in unselected, traditionally defined TIAs. Applying this model to the US population in the year 2000 showed that adopting a tissue-based definition of TIA would decrease the annual number of events classified as TIAs from 179,840 to 120,493 and increase events classified as strokes from 821,181 to 880,520. CONCLUSIONS: Adopting a tissue-based definition of transient ischemic attack would reduce estimates of the annual incidence of TIA by 33% (sensitivity analysis range, 19% to 44%) and increase estimates of the annual incidence of stroke in the United States by 7% (range, 4% to 10%). PMID- 12637704 TI - Hand-in-hand in the "trading zone". PMID- 12637702 TI - Platelet function under aspirin, clopidogrel, and both after ischemic stroke: a case-crossover study. AB - BACKGROUND AND PURPOSE: Combined antiplatelet agents may offer additive protection over single drugs after stroke. We investigated whether platelet activation is reduced under combined aspirin and clopidogrel compared with each drug alone. METHODS: In a case-crossover study, 31 patients with previous atherothrombotic or lacunar stroke who were treated with aspirin (100 to 300 mg/d) received clopidogrel (75 mg/d) and both aspirin and clopidogrel for 4 weeks. Platelet function in whole blood was studied after each treatment period and in healthy control subjects to assess activation-dependent antigens CD62p and CD63 by flow cytometry and collagen/epinephrine (CEPI-CT) and collagen/ADP (CADP CT) closure times with the platelet function analyzer PFA-100, which investigates platelet-related function under shear stress. RESULTS: CD62p expression and CD63 expression were not different under the 3 treatment regimens. CD63 but not CD62p expression was lower in control subjects than in stroke patients regardless of the antiplatelet treatment (P<0.05). CEPI-CT was prolonged under aspirin and aspirin plus clopidogrel compared with clopidogrel monotherapy (P<0.0001). CADP CT was longer under combination therapy than under aspirin (P=0.0009) or clopidogrel (P=0.0074) or in control subjects (P=0.0010), mainly because of strong prolongation in a patient subgroup (28%). CONCLUSIONS: CD63 expression reflecting the release of platelet lysosomes is consistently increased after stroke and incompletely suppressed by treatment with aspirin, clopidogrel, or both. The strong prolongation of CADP-CT under combined aspirin and clopidogrel in a patient subgroup may indicate a lower risk of thrombosis but also a higher risk of hemorrhage. The predictive value of platelet activation parameters requires investigation in prospective studies. PMID- 12637706 TI - Space science. Star-crossed comet chaser eyes new target. PMID- 12637705 TI - Intellectual property. Judge turns Rochester's golden patent into lead. PMID- 12637707 TI - Infectious diseases. Resistant staph finds new niches. PMID- 12637708 TI - Metrology. A thermometer beyond compare? PMID- 12637709 TI - Functional genomics. How to make sense of sequence. PMID- 12637710 TI - Arctic ecology. For precarious populations, pollutants present new perils. PMID- 12637711 TI - Environmental protection. Pentagon wants out from 'green' rules. PMID- 12637712 TI - Global change. Who pushed whom out of the last ice age? PMID- 12637714 TI - Astronomy. Liquid-mirror telescope set to give stargazing a new spin. PMID- 12637713 TI - Psychopharmacology. The medication merry-go-round. PMID- 12637716 TI - Archaeology. Impending war, dam hinder Iraqi preservation efforts. PMID- 12637715 TI - Clinical trials. New rule triggers debate over best way to test drugs. PMID- 12637717 TI - Ophthalmology. Tailor-made vision descends to the eye of the beholder. PMID- 12637718 TI - Ophthalmology. Coming soon: 'wavefront eye surgery'? PMID- 12637720 TI - Another look at NEAR. PMID- 12637719 TI - Research cloning, ethics, and public policy. PMID- 12637721 TI - Structural genomics: current progress. PMID- 12637722 TI - Climate change. Making deforestation pay under the Kyoto Protocol? PMID- 12637724 TI - Physics. Topology from the bottom up. PMID- 12637725 TI - Geocryology. (Un)frozen in time. PMID- 12637723 TI - Virology. Forced entry--or does HTLV-I have the key? PMID- 12637726 TI - Geophysics. The thermal state of Earth's core. PMID- 12637727 TI - Cell biology. A matter of life or death. PMID- 12637728 TI - Cancer. Developing molecular biomarkers for cancer. PMID- 12637731 TI - Connecting the dots to custom catalysts. PMID- 12637729 TI - Evolution. Little else but parasites. PMID- 12637732 TI - Water splitting goes au naturel. PMID- 12637733 TI - The impact of nanoscience on heterogeneous catalysis. AB - Most catalysts consist of nanometer-sized particles dispersed on a high-surface area support. Advances in characterization methods have led to a molecular-level understanding of the relationships between nanoparticle properties and catalytic performance. Together with novel approaches to nanoparticle synthesis, this knowledge is contributing to the design and development of new catalysts. PMID- 12637734 TI - Privileged chiral catalysts. AB - One of the most active current areas of chemical research is centered on how to synthesize handed (chiral) compounds in a selective manner, rather than as mixtures of mirror-image forms (enantiomers) with different three-dimensional structures (stereochemistries). Nature points the way in this endeavor: different enantiomers of a given biomolecule can exhibit dramatically different biological activities, and enzymes have therefore evolved to catalyze reactions with exquisite selectivity for the formation of one enantiomeric form over the other. Drawing inspiration from these natural catalysts, chemists have developed a variety of synthetic small-molecule catalysts that can achieve levels of selectivity approaching, and in some cases matching, those observed in enzymatic reactions. PMID- 12637735 TI - Dispelling the myths--biocatalysis in industrial synthesis. AB - Biocatalysis has emerged as an important tool in the industrial synthesis of bulk chemicals, pharmaceutical and agrochemical intermediates, active pharmaceuticals, and food ingredients. However, the number and diversity of the applications are modest, perhaps in part because of perceived or real limitations of biocatalysts, such as limited enzyme availability, substrate scope, and operational stability. Recent scientific breakthroughs in genomics, directed enzyme evolution, and the exploitation of biodiversity should help to overcome these limitations. As a result, we expect many new industrial applications of biocatalysis to be realized, from single-step enzymatic conversions to customized multistep microbial synthesis by means of metabolic pathway engineering. PMID- 12637736 TI - Catalytic nanoarchitectures--the importance of nothing and the unimportance of periodicity. AB - Heterogeneous catalysis has always been an inherently nanoscopic phenomenon with important technological and societal consequences for energy conversion and the production of chemicals. New opportunities for improved performance arise when the multifunctionality inherent in catalytic processes, including molecular transport of reactants and products, is rethought in light of architectures designed and fabricated from the appropriate nanoscale building blocks, including the use of "nothing" (void space) and deliberate disorder as design components. Architectures with all of the appropriate electrochemical and catalytic requirements, including large surface areas readily accessible to molecules, may now be assembled on the benchtop. Designing catalytic nanoarchitectures that depart from the hegemony of periodicity and order offers the promise of even higher activity. PMID- 12637737 TI - Homogeneous catalysis--new approaches to catalyst separation, recovery, and recycling. AB - Homogeneous catalysts have many attractive properties, such as high selectivities. However, many homogeneous catalytic systems cannot be commercialized because of difficulties associated with separating the products from the catalyst. Recent approaches to tackling this problem are reviewed and compared. PMID- 12637738 TI - Ethnobotanical knowledge shared widely among Tsimane' Amerindians, Bolivia. PMID- 12637739 TI - Meltwater pulse 1A from Antarctica as a trigger of the Bolling-Allerod warm interval. AB - Meltwater pulse 1A (mwp-1A) was a prominent feature of the last deglaciation, which led to a sea-level rise of approximately 20 meters in less than 500 years. Concurrent with mwp-1A was the onset of the Bolling-Allerod interstadial event (14,600 years before the present), which marked the termination of the last glacial period. Previous studies have been unable to reconcile a warm Northern Hemisphere with mwp-1A originating from the Laurentide or Fennoscandian ice sheets. With the use of a climate model of intermediate complexity, we demonstrate that with mwp-1A originating from the Antarctic Ice Sheet, consistent with recent sea-level fingerprinting inferences, the strength of North Atlantic Deep Water (NADW) formation increases, thereby warming the North Atlantic region and providing an explanation for the onset of the Bolling-Allerod warm interval. The established mode of active NADW formation is then able to respond to subsequent freshwater forcing from the Laurentide and Fennoscandian ice sheets, setting the stage for the Younger Dryas cold period. PMID- 12637740 TI - Grain boundary scars and spherical crystallography. AB - We describe experimental investigations of the structure of two-dimensional spherical crystals. The crystals, formed by beads self-assembled on water droplets in oil, serve as model systems for exploring very general theories about the minimum-energy configurations of particles with arbitrary repulsive interactions on curved surfaces. Above a critical system size we find that crystals develop distinctive high-angle grain boundaries, or scars, not found in planar crystals. The number of excess defects in a scar is shown to grow linearly with the dimensionless system size. The observed slope is expected to be universal, independent of the microscopic potential. PMID- 12637741 TI - Epitaxial BiFeO3 multiferroic thin film heterostructures. AB - Enhancement of polarization and related properties in heteroepitaxially constrained thin films of the ferroelectromagnet, BiFeO3, is reported. Structure analysis indicates that the crystal structure of film is monoclinic in contrast to bulk, which is rhombohedral. The films display a room-temperature spontaneous polarization (50 to 60 microcoulombs per square centimeter) almost an order of magnitude higher than that of the bulk (6.1 microcoulombs per square centimeter). The observed enhancement is corroborated by first-principles calculations and found to originate from a high sensitivity of the polarization to small changes in lattice parameters. The films also exhibit enhanced thickness-dependent magnetism compared with the bulk. These enhanced and combined functional responses in thin film form present an opportunity to create and implement thin film devices that actively couple the magnetic and ferroelectric order parameters. PMID- 12637742 TI - Recent trends in Arctic surface, cloud, and radiation properties from space. AB - Trends in satellite-derived cloud and surface properties for 1982 to 1999 show that the Arctic has warmed and become cloudier in spring and summer but has cooled and become less cloudy in winter. The increase in spring cloud amount radiatively balances changes in surface temperature and albedo, but during summer, fall, and winter, cloud forcing has tended toward increased cooling. This implies that, if seasonal cloud amounts were not changing, surface warming would be even greater than that observed. Strong correlations with the Arctic Oscillation indicate that the rise in surface temperature and changes in cloud amount are related to large-scale circulation rather than to local processes. PMID- 12637743 TI - Timing of atmospheric CO2 and Antarctic temperature changes across termination III. AB - The analysis of air bubbles from ice cores has yielded a precise record of atmospheric greenhouse gas concentrations, but the timing of changes in these gases with respect to temperature is not accurately known because of uncertainty in the gas age-ice age difference. We have measured the isotopic composition of argon in air bubbles in the Vostok core during Termination III (approximately 240,000 years before the present). This record most likely reflects the temperature and accumulation change, although the mechanism remains unclear. The sequence of events during Termination III suggests that the CO2 increase lagged Antarctic deglacial warming by 800 +/- 200 years and preceded the Northern Hemisphere deglaciation. PMID- 12637744 TI - Climate and the collapse of Maya civilization. AB - In the anoxic Cariaco Basin of the southern Caribbean, the bulk titanium content of undisturbed sediment reflects variations in riverine input and the hydrological cycle over northern tropical South America. A seasonally resolved record of titanium shows that the collapse of Maya civilization in the Terminal Classic Period occurred during an extended regional dry period, punctuated by more intense multiyear droughts centered at approximately 810, 860, and 910 A.D. These new data suggest that a century-scale decline in rainfall put a general strain on resources in the region, which was then exacerbated by abrupt drought events, contributing to the social stresses that led to the Maya demise. PMID- 12637745 TI - Evolution of virulence in a plant host-pathogen metapopulation. AB - In a wild plant-pathogen system, host resistance and pathogen virulence varied markedly among local populations. Broadly virulent pathogens occurred more frequently in highly resistant host populations, whereas avirulent pathogens dominated susceptible populations. Experimental inoculations indicated a negative trade-off between spore production and virulence. The nonrandom spatial distribution of pathogens, maintained through time despite high pathogen mobility, implies that selection favors virulent strains of Melampsora lini in resistant Linum marginale populations and avirulent strains in susceptible populations. These results are consistent with gene-for-gene models of host pathogen coevolution that require trade-offs to prevent pathogen virulence increasing until host resistance becomes selectively neutral. PMID- 12637746 TI - Rapid evolution of egg size in captive salmon. AB - Captive breeding and release programs, widely used to supplement populations of declining species, minimize juvenile mortality to achieve rapid population growth. However, raising animals in benign environments may promote traits that are adaptive in captivity but maladaptive in nature. In chinook salmon, hatchery rearing relaxes natural selection favoring large eggs, allowing fecundity selection to drive exceptionally rapid evolution of small eggs. Trends toward small eggs are also evident in natural populations heavily supplemented by hatcheries, but not in minimally supplemented populations. Unintentional selection in captivity can lead to rapid changes in critical life-history traits that may reduce the success of supplementation or reintroduction programs. PMID- 12637747 TI - Modulating sphingolipid biosynthetic pathway rescues photoreceptor degeneration. AB - Mutations in proteins of the Drosophila phototransduction cascade, a prototypic guanine nucleotide-binding protein-coupled receptor signaling system, lead to retinal degeneration and have been used as models to understand human degenerative disorders. Here, modulating the sphingolipid biosynthetic pathway rescued retinal degeneration in Drosophila mutants. Targeted expression of Drosophila neutral ceramidase rescued retinal degeneration in arrestin and phospholipase C mutants. Decreasing flux through the de novo sphingolipid biosynthetic pathway also suppressed degeneration in these mutants. Both genetic backgrounds modulated the endocytic machinery because they suppressed defects in a dynamin mutant. Suppression of degeneration in arrestin mutant flies expressing ceramidase correlated with a decrease in ceramide levels. Thus, enzymes of sphingolipid metabolism may be suitable targets in the therapeutic management of retinal degeneration. PMID- 12637748 TI - Dissecting temporal and spatial control of cytokinesis with a myosin II Inhibitor. AB - Completion of cell division during cytokinesis requires temporally and spatially regulated communication from the microtubule cytoskeleton to the actin cytoskeleton and the cell membrane. We identified a specific inhibitor of nonmuscle myosin II, blebbistatin, that inhibited contraction of the cleavage furrow without disrupting mitosis or contractile ring assembly. Using blebbistatin and other drugs, we showed that exit from the cytokinetic phase of the cell cycle depends on ubiquitin-mediated proteolysis. Continuous signals from microtubules are required to maintain the position of the cleavage furrow, and these signals control the localization of myosin II independently of other furrow components. PMID- 12637749 TI - Protein insertion into the mitochondrial inner membrane by a twin-pore translocase. AB - The mitochondrial inner membrane imports numerous proteins that span it multiple times using the membrane potential Deltapsi as the only external energy source. We purified the protein insertion complex (TIM22 complex), a twin-pore translocase that mediated the insertion of precursor proteins in a three-step process. After the precursor is tethered to the translocase without losing energy from the Deltapsi, two energy-requiring steps were needed. First, Deltapsi acted on the precursor protein and promoted its docking in the translocase complex. Then, Deltapsi and an internal signal peptide together induced rapid gating transitions in one pore and closing of the other pore and drove membrane insertion to completion. Thus, protein insertion was driven by the coordinated action of a twin-pore complex in two voltage-dependent steps. PMID- 12637750 TI - Loss of IGF2 imprinting: a potential marker of colorectal cancer risk. AB - Loss of imprinting (LOI), an epigenetic alteration affecting the insulin-like growth factor II gene (IGF2), is found in normal colonic mucosa of about 30% of colorectal cancer (CRC) patients, but it is found in only 10% of healthy individuals. In a pilot study to investigate the utility of LOI as a marker of CRC risk, we evaluated 172 patients at a colonoscopy clinic. The adjusted odds ratio for LOI in lymphocytes was 5.15 for patients with a positive family history [95% confidence interval (95% CI), 1.70 to 16.96; probability P = 0.002], 3.46 for patients with adenomas (95% CI, 1.14 to 11.37; P = 0.026), and 21.7 for patients with CRC (95% CI, 3.48 to 153.6; P = 0.0005). LOI can be assayed with a DNA-based blood test, and it may be a valuable predictive marker of an individual's risk for CRC. PMID- 12637751 TI - Bovine adenovirus type 3 pneumonia in dexamethasone-treated calves. AB - The effects of immunosuppression were examined in 1.5-month-old calves that were given dexamethasone (DM) before endobronchial inoculation with bovine adenovirus type 3 (BAV-3). Immunohistopathologically, severe necrotizing bronchiolitis with eosinophilic and basophilic intranuclear inclusion bodies was observed both in DM treated 1.5-month-old infected calves and in non-DM-treated 7-day-old infected calves. These inclusion bodies were correlated with the detection of BAV-3 antigen and viral particles. The presence of inclusion bodies in the desquamated epithelial cells or of BAV-3 antigen, or both, correlated well with the isolated level of BAV-3 in bronchoalveolar lavage (BAL) fluid. Few immunoglobulin (IgG, IgM, and IgA)-containing B lymphocytes or CD8+ T lymphocytes infiltrated the pneumonic lesion in both the 7-day-old and the DM-treated 1.5-month-old infected calves. Thus, depletion of CD8+ T lymphocytes in calves might influence the clearance of BAV-3 from respiratory tissues. PMID- 12637752 TI - Adrenal cortical carcinomas with myxoid differentiation in the domestic ferret (Mustela putorius furo). AB - A total of 15 adrenocortical carcinomas with myxoid differentiation from 15 ferrets were evaluated in this retrospective study. Six of these ferrets (40%) either were euthanatized or died due to invasive and/or metastatic disease. The myxoid component was a variable part (between 5% and 95%) of the adrenal cortical neoplasm and consisted of sheets and cords of small, polygonal neoplastic cells that formed lumenlike spaces. Such spaces contained a variable amount of alcian blue (pH 2.5)-positive mucinous product (i.e., acidic mucopolysaccharides). Neoplastic cells were negative for the argentaffin reaction, but immunohistochemically they were strongly positive for vimentin and alpha-inhibin and lightly positive for synaptophysin. Proliferating cell nuclear antigen (PCNA) labeling indices (LI) of adrenal cortical neoplastic cells within the myxoid component of the neoplasm were significantly elevated (P < 0.05) compared with those of typical neoplastic adrenal cortical cells or the adjacent nonneoplastic zona reticularis. Ultrastructurally, cells in the myxoid component exhibited a typical adrenocortical phenotype characterized by cytoplasmic lipid vacuoles, prominent rough and smooth endoplasmic reticulum, and zonula adherens. This lesion was interpreted as an adrenal cortical carcinoma with myxoid differentiation and appeared to be highly malignant based on PCNA LI, rate of invasion into adjacent tissue, and metastasis (6/15). This report is the first description of this histologic variant in the ferret, which morphologically resembled the rare myxoid variant of adrenocortical carcinoma described in humans. PMID- 12637753 TI - Detection rates of porcine reproductive and respiratory syndrome virus, porcine circovirus type 2, and swine influenza virus in porcine proliferative and necrotizing pneumonia. AB - A retrospective study on pig lung tissues from 60 cases of proliferative and necrotizing pneumonia (PNP) was performed to determine the presence of porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), and porcine circovirus type 2 (PCV2) in these lesions. Cases selected included 30 cases diagnosed between 1988 and 1992 and 30 cases diagnosed between 1997 and 2001. In each group of 30 cases, 10 were from suckling piglets, whereas the other 20 were from postweaned animals representing either nursery or grower-finisher pigs. Immunohistochemistry using a monoclonal antibody to influenza virus type A was used to determine the presence of SIV, and in situ hybridization was used for the detection of PRRSV and PCV2 nucleic acids. PRRSV was detected in 55 of the 60 cases examined (92%), PCV2 in 25 cases (42%), and SIV in only 1 case (2%). In 30 cases (50%), PRRSV was the only virus detected, whereas in 25 other cases (42%), a combination of PRRSV and PCV2 could be detected in the lungs with PNP lesions. PCV2 could not be detected in the lungs of suckling pigs with PNP. All PCV2 positive cases were found in postweaned pigs and were always in combination with PRRSV. In this latter age group, PCV2 was detected in 63% of the cases (25/40). Data from our study indicate that SIV is rarely identified in PNP and that PCV2 infection is not essential for the development of PNP lesions. The results of the present study demonstrate that PRRSV is consistently and predominantly associated with PNP and should be considered the key etiologic agent for the condition. PMID- 12637754 TI - Ehrlichia platys (Anaplasma platys) in dogs from Maracaibo, Venezuela: an ultrastructural study of experimental and natural infections. AB - Since 1982 Ehrlichia platys, now emended as Anaplasma platys, has been diagnosed in dogs from Maracaibo, Venezuela, using buffy coat smears stained with Dip Quick. Three dogs were inoculated with an A. platys strain. When parasitemia reached 60-97%, blood samples obtained from the inoculated dogs and from two naturally infected dogs were centrifuged to obtain platelet-rich plasma, which was mixed with 0.1% glutaraldehyde at 37 C for 10 minutes. Platelet pellets were fixed in 3% glutaraldehyde for 72 hours and processed for conventional transmission electron microscopy. Platelets contained pleomorphic organisms with a distinct double membrane that was not observed when the bodies were in a determinate developmental stage. There were 1-15 individual bodies included in a host cell vacuole. The organisms had an electron-lucent inner area, whereas the internal surface of their inner plasma membranes exhibited an electron-dense rough substance. In naturally infected dogs, organisms with different ultrastructural features were found inside the same platelet. Some organisms contained central dense material surrounded by a pale zone, which was in turn surrounded by a moderately dense peripheral area. Other organisms contained an eccentrically electron-dense material. The intravacuolar space appeared fully electron-lucent. Each organism usually exhibited inner fine strands. Empty structures displaying junctions with the vacuolar membrane were observed. Our results indicate that distinct ultrastructural characteristics are associated with different stages of A. platys development and may differ among A. platys strains. PMID- 12637755 TI - Classical Swine Fever: pathology of bone marrow. AB - Twenty pigs were inoculated with a virulent isolate (Quillota strain) of classical swine fever (CSF) virus to determine the chronological development of lesions in bone marrow. Histopathologic, ultrastructural and immunohistochemical (detection of viral antigen gp55, myeloid-histiocyte antigen, CD3 antigen, and FVIII-rag), and morphometric techniques were employed. Viral antigen was detected from 2 days postinfection (dpi) in stromal and haematopoitic cells, and severe atrophy related to apoptosis of haematopoitic cells was observed. Megakaryocytes (MKs) did not show significant changes in number, but there were important qualitative changes including 1) increased numbers of cloud-nuclei MKs, microMKs, apoptotic MKs, and atypical nucleated MKs and 2) decreased number of typical nucleated MKs. Morphometric study of these cells showed a decrease in cytoplasmic area. MK infection was detected from 2 dpi, but in a small percentage of cells. Myeloid cells showed quantitative changes, with an increase in granulocyte numbers. Apoptosis of lymphocytes and viral infection of erythroblasts were also observed. The main changes in stroma were depletion of T lymphocytes in the middle phase of the experiment and macrophages. Viral infection was also observed in these cells. MK lesions suggest dysmegakaryocytopoiesis, which would aggravate the thrombocytopenia already present and could be responsible for it. Granulocyte changes would lead to the appearance of circulating immature forms, whereas lymphocyte apoptosis in bone marrow would contribute to lymphopenia. PMID- 12637756 TI - Accumulation of pathogenic prion protein (PrPSc) in nervous and lymphoid tissues of sheep with subclinical scrapie. AB - All sheep older than 1 year of age from a flock of the Rygja breed in which clinical scrapie was detected for the first time in two animals (4%) were examined for accumulation of pathogenic prion protein (PrPSc) by immunohistochemistry in the obex, the cerebellum, and the medial retrophayngeal lymph node. In addition, six lambs, 2-3 months old, all offspring of PrPSc positive dams, were examined for PrPSc in the ileal Peyers' patch (IPP), the distal jejunal lymph node, the spleen, and the medial retropharyngeal lymph node (RPLN). In this flock, 35% (17/48) of the adult sheep showed accumulation of PrPSc, an eightfold increase compared with clinical disease. All positives carried susceptible PrP genotypes. Three sheep had deposits of PrPSc in the RPLN and not in the brain, suggesting that this organ, easily accessible at slaughter, is suitable for screening purposes. Two 7-year-old clinically healthy homozygous V136Q171 ewes showed sparse immunostaining in the central nervous system and may have been infected as adults. Further, two littermates, 86-days-old, showed PrPSc in the IPP. Interestingly, one of these lambs had the intermediate susceptible PrP genotype, VA136QR171. In addition to early immunolabeling in the dorsal motor nucleus of the vagal nerve, a few of the sheep had early involvement of the cerebellum. In fact, a 2-year-old sheep had sparse deposits of PrPSc in the cerebellum only. Because experimental bovine spongiform encephalopathy (BSE) in sheep seems to behave in a similar manner as natural scrapie, these results, particularly regarding spread of infectivity, may have implications for the handling of BSE should it be diagnosed in sheep. PMID- 12637757 TI - Histology of uterine leiomyoma and occurrence in relation to reproductive activity in the Baltic gray seal (Halichoerus grypus). AB - A high prevalence of uterine leiomyoma has been reported in Baltic gray seals aged 15 years and above. Studies on Baltic seals during the 1970s revealed high tissue concentrations of the organochlorines bis(chlorophenyl)-1,1,1 trichloroethane (DDT) and polychlorinated biphenyls (PCBs), lowered reproduction rate, and pathologic changes. In the second half of the 1970s, decreases of PCB and DDT in Baltic biota occurred, and the prevalence of pregnancies in Baltic seals increased. Between 1975 and 1997, 53 Baltic gray seal females of age 15-40 years were found dead and sent to the Swedish Museum of Natural History. Seals were autopsied and 34/53 (64%) had uterine leiomyomas. Samples from 15 were sufficiently well preserved for histologic examination. Uterine leiomyomas were found most commonly in the uterine corpus but also were observed in the uterine horns, cervix, and vagina. Cut surfaces of the leiomyomas appeared as whorled white fibrous tissue. Histologically, spindle cells were arranged in a whorl-like pattern. The nuclei were rod-like and strikingly uniform in shape and size. Mitotic figures were rare. Immunohistochemical staining of the tumors showed a positive reaction to antibodies recognizing smooth muscle actin. Reproductively active gray seals have an ovarian corpus luteum or albicans for most of the year. In 22/34 (65%) gray seals with uterine leiomyomas, ovaries did not contain corpora. In gray seals without macroscopically detected uterine leiomyoma, ovaries from 6/19 (32%) seals had no corpora. It is possible that the development of leiomyoma in the seals is associated with organochlorines and the previous low reproductive activity. PMID- 12637758 TI - Expression of monocyte chemoattractant protein-1 but not interleukin-8 in granulomatous lesions in lymph nodes from pigs with naturally occurring postweaning multisystemic wasting syndrome. AB - Monocyte chemoattractant protein-1 (MCP-1) but not interleukin-8 (IL-8) was detected by in situ hybridization using a nonradioactive digoxigenin-labeled complementary DNA probe in granulomatous lesions of lymph nodes from 20 pigs with naturally occurring postweaning multisystemic wasting syndrome (PMWS). Complementary DNA probes of 375 and 266 base pairs for MCP-1 and IL-8, respectively, were generated by reverse transcription-polymerase chain reaction. The 20 pigs with PMWS had distinct positive hybridization signals for MCP-1 but not for IL-8. The hybridization signals for MCP-1 were strictly confined to the cells with granulomatous lesions, including macrophages and multinucleated giant cells. A very close cell-to-cell correlation between MCP-1 and porcine circovirus 2 was seen in serial sections of lymph nodes. Results of this study indicate that MCP-1 expression may play a role in the pathogenesis of granulomatous inflammation in pigs with PMWS. PMID- 12637759 TI - Renal lesions in spontaneous insulin-dependent diabetes mellitus in the nonobese diabetic mouse: acute phase of diabetes. AB - The nonobese diabetic mouse is a model of spontaneous insulin-dependent diabetes mellitus. The present study made longitudinal observations of renal lesions in the acute-progressive phase of diabetic mice 0, 10, 20, 30, and 40 days after onset of diabetes without insulin therapy. Plasma creatinine and blood urea nitrogen concentrations gradually increased after onset of diabetes. Kidney weight increased and plateaued at day 20. Under electron microscopy the glomeruli demonstrated only mild changes on day 40. In the proximal tubules proliferating cell nuclear antigen-positive nuclei and nuclear divisions were increased on days 10 and 20. On day 40 of diabetes, increased periodic acid-Schiff-positive granules, confirmed as lysosomal dense bodies, increased neuronal nitric oxide synthase (nNOS) positive reaction, and decreased periodic acid-Schiff staining in the brush border were observed in the proximal straight tubules. In the juxtaglomerular apparatus stratified macula densa were decreased with time in diabetes compared with the findings on day 0, and this macula densa positively reacted with nNOS. No changes in renin levels were observed. In addition, apoptotic cells were not detected. In conclusion, this research represents the first thorough characterization of acute changes in nonobese diabetic mouse kidneys. The results demonstrated renal hypertrophy and slight glomerular injury in early stages and structural alteration of the proximal straight tubules at later stages during the acute phase of diabetes. Furthermore, increased nNOS may represent one of the pathogenic factors of diabetic nephropathy. PMID- 12637760 TI - Immunohistochemical study of retinol-binding protein in livers of polar bears (Thalarctos maritimus). AB - Liver tumors of unknown cause have frequently been described in polar bears. Concurrent decrease of vitamin A levels and chronic liver disease are associated with hepatic carcinogenesis in humans. More than 90% of the body's vitamin A is stored in the liver, where it is bound to an intracellular retinol-binding protein (RBP). Therefore, in this retrospective study, RBP was assessed by immunohistochemistry in liver sections of 11 polar bears. Two of these polar bears had hepatocellular carcinoma, four showed other chronic liver changes, and five had normal livers. In normal livers, the cytoplasm stained diffusely positive with intensely staining cytoplasmic granules. RBP staining was evaluated and the abundance of diffuse cytoplasmic staining and intracytoplasmic large granules was determined. All cases with pathologic liver changes had markedly decreased staining intensities for RBP compared with normal livers. The findings of this study suggest that in polar bears, as in humans, vitamin A metabolism may play a role in hepatic carcinogenesis. PMID- 12637761 TI - Parathyroid gland carcinoma in a Wistar rat. AB - An anaplastic carcinoma was found in one of the two parathyroids of a 2-year-old male Wistar rat, which was sacrificed at the end of a carcinogenicity study. Morphologically, it was characterized by the presence of nodular areas of pleomorphic and dense cells with numerous atypical mitoses and large regions of smaller and dark monomorphic cells devoid of mitoses and forming small cystic spaces. Local invasion of the capsule and pronounced compression of the parenchyma of the thyroid gland were observed. Immunohistochemically, the tumor was markedly positive for the parathyroid hormone and negative for the thyroid transcription factor. The proliferative activity was assessed by immunostaining the endogenous cell proliferation associated-antigen Ki-67, and the proliferating cell nuclear antigen. The diagnosis of carcinoma of the parathyroid was made on the basis of microscopic and immunohistochemical findings. PMID- 12637762 TI - Cyclooxygenase-2 expression in canine mammary tumors. AB - Mammary tumors are the most common neoplasms in female dogs. Induction of cyclooxygenase-2 (COX-2) has been implicated in various cancers in humans. However, expression of COX-2 has not been investigated in canine mammary tumors. Normal mammary gland (n = 4), simple or complex adenomas (n = 63), and simple or complex adenocarcinomas (n = 84) were studied by immunohistochemistry. Results showed that COX-2 was not expressed in the normal gland but was detected in 24% of adenomas and in 56% of adenocarcinomas (P < 0.001). The incidence of COX-2 expression and the intensity of the COX-2 signal were higher in adenocarcinomas than in adenomas (P < 0.001). These results demonstrate for the first time that COX-2 is induced in a proportion of canine mammary tumors and that COX-2 expression is more frequent and more intense in malignant than in benign tumors, suggesting a potential role for COX-2 in canine mammary tumorigenesis. PMID- 12637763 TI - Immunohistochemical evaluation of a malignant intestinal carcinoid in a dog. AB - An intestinal carcinoid with multiple metastases was identified in a 5-year-old male Shih Tzu with a clinical history of anemia, fatigue, anorexia, vomiting, intermittent diarrhea, intestinal bleeding, and progressive emaciation. There was a yellowish-white mass 15 mm in diameter in the anterior jejunum and white nodules consistent with metastases in many organs. Histopathologically, the mass consisted of neoplastic cells arranged in lobules, trabeculae, or closely interdigitating islands of cells. Neoplastic cells were generally polygonal with round hyperchromatic nuclei, modest amounts of eosinophilic cytoplasm, and eosinophilic cytoplasmic granules. Mitoses were common. Rosette formations of tumor cells were apparent in metastatic tumors. Immunohistochemically, tumor cells stained positive for cytokeratin 13, synaptophysin, protein gene product 9.5, neuron-specific enolase, chromogranin A, calcitonin gene-related peptide, serotonin (5-HT), and Leu-7. Serum 5-HT concentrations for this dog were increased 10-fold compared with those of normal dogs. All findings were consistent with a diagnosis of a malignant intestinal carcinoid. PMID- 12637764 TI - Brodifacoum toxicosis in two neonatal puppies. AB - Eight out of a litter of 13 puppies were either born dead or died within 48 hours of birth. Three puppies that died shortly after birth were necropsied. Two puppies had hemorrhage in the thoracic and peritoneal cavities, intestinal serosa, and meninges. The third puppy was smaller than the other two puppies but did not have detectable hemorrhage. Brodifacoum, a second-generation coumarin anticoagulant, was detected in livers from the two puppies with hemorrhage. The dam did not have clinical signs of coagulopathy before or subsequent to whelping. The owners were confident that the dog had not been exposed to rodenticide for at least 4 weeks before whelping. A presumptive diagnosis of in utero brodifacoum toxicity was made. To the authors' knowledge this is the first time a second generation coumarin anticoagulant has been detected in the liver of a newborn animal. This case is also unique because the dam was unaffected, suggesting that fetuses are more susceptible to brodifacoum toxicity than adult animals. PMID- 12637765 TI - Quantitative analysis of tryptase- and chymase-containing mast cells in eosinophilic conditions of cats. AB - The presence and density of tryptase-positive/chymase-positive mast cells (MCs) (MC(TC)), chymase-positive/tryptase-negative MCs (MCC), and tryptase positive/chymase-negative MCs (MC(T)) in lesional skin from cats with eosinophilic conditions were investigated. Skin biopsy specimens from eight cats with eosinophilic plaque (three cats), eosinophilic granuloma (two cats), and eosinophilic dermatitis (three cats) were studied. Toluidine blue staining and a double-enzyme-immunohistochemical staining technique were performed to determine MC density and MC subtypes, respectively. MC density varied from 170.3 to 503 cells/mm2 (mean value of 314.9 cells/mm2). In the superficial dermis, 5.9% of the MC belonged to the MC(T), 12.8% to the MC(C), and 81.2% to the MC(TC) subtype. In the deep dermis, 12.8% belonged to the MC(T), 12.8% to the MC(C), and 73.8% to the MC(TC) subtype. It is the first time that MC(C) have been identified. The double-labeling procedure proved to be a reliable tool for identifying simultaneously the presence of MC subtypes in feline skin. PMID- 12637767 TI - Cell cycle control mechanisms in B-1 and B-2 lymphoid subsets. AB - An effective humoral response requires that a given B lymphocyte population express a repertoire of receptors capable of recognizing a distinct array of antigens, while at the same time disregarding self-antigens. Mature B cells interacting with antigen via their B cell antigen receptors (BCRs) enter G(1) phase of the cell cycle and, depending on the strength of the signal, can commit to S phase entry. Input from co-receptors, which may function to either enhance or inhibit BCR signals, also influence the decision to proliferate. We review herein recent advances in the biochemistry of G(1)-cyclin holoenzymes that function to integrate BCR-coupled signaling pathways to the phosphorylation (and inactivation) of the retinoblastoma gene product (pRb) in splenic B lymphocytes (B-2 cells). We also highlight differences in the control of G(1)-to-S phase progression between B-2 cells and peritoneal CD5+ B cells (B-1 cells). PMID- 12637768 TI - Immunity to liver stage malaria: considerations for vaccine design. AB - Malaria is the world's deadliest parasitic disease and effective control measures are a public health priority. Most deaths in humans from malaria are caused by one species of the protozoa, Plasmodium falciparum. An efficacious and cost effective vaccine against this parasite is considered a holy grail of modern molecular medicine. A vaccine that targets liver stage parasites would prevent infection from reaching the blood and causing clinical disease. Among known P. falciparum antigens, liver stage antigen-1 (LSA-1) is the only protein expressed exclusively by infected hepatocytes. Several independent studies in humans have consistently related immune responses to LSA-1 with resistance to infection or disease, providing a powerful rationale for the development of liver stage vaccines. Investigations by ourselves and others aim to dissect the mechanism of cellular immunity to LSA-1 and to evaluate in different delivery systems epitopes associated with protection as components of a multiantigen malaria vaccine. The first clinical trials are already being conducted, the results of which are eagerly awaited. PMID- 12637766 TI - Expression of nonclassical MHC class Ib genes: comparison of regulatory elements. AB - Peptide binding proteins of the major histocompatibility complex consist of the "classical" class Ia and "nonclassical" class Ib genes. The gene organization and structure/function relationship of the various exons comprising class I proteins are very similar among the class Ia and class Ib genes. Although the tissue specific patterns of expression of these two gene families are overlapping, many class Ib genes are distinguished by relative low abundance and/or limited tissue distribution. Further, many of the class Ib genes serve specialized roles in immune responses. Given that the coding sequences of the class Ia and class Ib genes are highly homologous we sought to examine the promoter regions of the various class Ib genes by comparison to the well characterized promoter elements regulating expression of the class Ia genes. This analysis revealed a surprising complexity of promoter structures among all class I genes and few instances of conservation of class Ia promoter regulatory elements among the class Ib genes. PMID- 12637769 TI - Diverse roles of integrins in human T lymphocyte biology. AB - T lymphocytes are the primary cells responsible for maintaining the immune system. There are many intricate mechanisms involved in the regulation of T cells and the integrin family of adhesive surface proteins plays a pivotal role in the control of T lymphocyte activation and functions. Integrins are heterodimeric transmembrane proteins that are not merely adhesion molecules but also function in T cell coactivation by providing a scaffold for signaling and cytoskeletal proteins that are adept at transmitting signals from the inside of the cell to the outside ("inside-out signaling") or from the outside of the cell to the inside ("outside-in signaling"). The signaling property of integrins allows for rapid responses to changes in the microenviroment of the lymphocyte. Therefore, whether the T cell needs to adhere or detach, integrins can quickly accommodate either state of the cell. Once cells are guided to sites of infection, inflammation, or antigen presentation, integrins can also participate in the initiation, maintenance, or termination of the response. This review will focus on the aspects of integrin-mediated T cell coactivation, affinity and avidity control of integrins, signaling molecules involved with integrins, association of integrins in lipid microdomains, and negative regulation of integrins. PMID- 12637772 TI - [Novel PKU treatment methods]. AB - Identification of a disease causing gene rarely leads to the invention of a novel therapy. Even in the case of genetic disorders or diseases greatly influenced by a hereditary component, knowledge about the genotype does not necessarily result in the invention of a novel therapy. In many cases effective treatment was introduced earlier without any existing knowledge of the molecular basis of the disease, or even without a full recognition of the pathomechanisms. Phenylketonuria is the best known example. Great expectation are connected with experimental therapy of phenylketonuria: enzymatic therapy, blocking the brain blood barrier for phenylalanine and novel forms of low phenylalanine diet. Molecular diagnostics could be helpful in identifying patients responsive to non standard therapy, such as tetrahydrobipterin supplementation. Novel research and development strategies will contribute to a reduction of the negative aspects of present treatment and they will facilitate normal physical a intellectual development of children with phenylketonuria, as well as to help patients in their everyday life. PMID- 12637770 TI - Novel mechanisms of class II major histocompatibility complex gene regulation. AB - Class II MHC molecules present processed peptides from exogenous antigens to CD4+ helper T lymphocytes. In so doing, they are central to immunity, driving both the humoral and cell mediated arms of the immune response. Class II MHC molecules, and the genes encoding them, are expressed primarily in cells of the immune system (B cells, thymic epithelial cells, activated T cells and professional antigen presenting cells). The expression is also under developmental control. Research over the past 20 years have provided a clear understanding of the cis elements and transcription factors that regulate the expression of Class II MHC genes. Perhaps the most critical advance has been the discovery of CIITA, a non- DNA binding activator of transcription that is a master control gene for class II gene expression. Current research is focused on understanding the situations where class II MHC gene expression occurs in a CIITA-independent pathway, and the molecular basis for this expression. Finally, significant emphasis is being placed on targeting class II MHC transcription factors to either inhibit or stimulate the immune response to transplanted tissue or in cell based vaccines. This communication outlines recent advances in this field and discusses likely areas for future research. PMID- 12637773 TI - [Atypical phenylketonuria treatment effectiveness]. AB - Atypical phenylketonuria is the consequence of the deficiency of tetrahydrobiopterin (BH4) - cofactor of phenylalanine hydroxylation. The type of the disease depends on the metabolic defect of synthesis or regeneration of the cofactor. The secondary deficiency of neurotransmitters - dopamine and serotonin is the reason of very severe clinical course and brain damage. Ten cases of atypical phenylketonuria are presented: 8 of the defect of 6-pyrogronyl tetrahydropterin synthase, 1 of dihydropteridine reductase, 1 of GTP cyclohydrolase. In 7 cases the diagnosis was confirmed by molecular studies. The age of the children at the time of diagnosis was from 3 weeks to 7 years. In 8 children pharmacological treatment was implemented without limitation of dietary intake of phenylalanine. Treatment period was from 9 months to 18.5 years. Treatment effectiveness is differentiated and independent of the age of diagnosis. Physical development in all but one patient is retarded and in all of them microcephaly is present. After treatment, mental development scores in 6 cases improved evidently (in 2 - up to normal values, in 4 - close to normal). No improvement was achieved in 2 cases (brothers). Detailed analysis of the clinical course in patients with similar age of diagnosis and start of treatment and with identical genotypes revealed significant differences in clinical course and treatment effectiveness. No explanation for this fact has been found. PMID- 12637774 TI - [Intrahepatic and peripheral blood NK cells in patients with chronic hepatitis C]. AB - The aim of this study was to evaluate the correlation between NK (natural killers) cells in an inflammatory infiltration of the liver and in peripheral blood in children with chronic hepatitis type C. MATERIAL AND METHODS: The study comprised 25 children with chronic hepatitis type C. The control group comprised 10 children with no liver disease in past medical history and normal activity of aminotransferases. In the study group, liver biopsy specimens were evaluated by histopathology and immunomorphology - the presence of NK cells present in the inflammatory infiltrate was determined. In all children the percentage and absolute count of NK cells in peripheral blood was evaluated by flow cytometry, as well as the activity of NK cells in relation to leukaemic cells of erythroleukemia K-562 without stimulation and after the stimulation with IL-2 (in vitro). RESULTS: In the studied group of children NK cells constituted 6.07 +/- 3.92 % of cells present in liver infiltrate. In peripheral blood NK cells constituted 11.68 +/- 6.73 % of white blood cells, and their absolute number was 241.08 +/- 128.56 /ml. The cytotoxic activity of NK cells in dilution E:T 25:1 was - 91.8 +/- 1.07 % and in dilution E:T 12.5:1 - 6.72 +/- 3.68 %. After stimulation with IL-2 it was 92.8 +/-1.01 % and 7.58 +/- 3.95 %, respectively. The number of NK cells in peripheral blood and cytotoxic activity of NK cells in the studied group did not differ from that of the control group. There was a positive correlation between the percentage of NK cells in the liver infiltrate and the absolute number of NK cells in the peripheral blood and between percentage of NK cells in the inflammatory infiltrate and cytotoxic activity of NK cells in peripheral blood, stimulated and non-stimulated with IL-2 in proportion E:T 12.5:1. CONCLUSIONS: Positive correlation between the number of NK cells in peripheral blood and liver infiltrate suggests the possibility of evaluating NK cells involvement in the pathogenesis of chronic hepatitis type C in children on the basis of peripheral blood tests. PMID- 12637771 TI - Signal transduction events regulating integrin function and T cell migration: new functions and complexity. AB - Integrin receptors facilitate T cell function by mediating adhesive events critical for T cell trafficking and recognition of foreign antigen, including interactions with vascular endothelium, extracellular matrix components, and antigen-presenting cells. Consequently, the functional activity of integrin receptors is acutely regulated by various intracellular signals delivered by other cell surface receptors, resulting in rapid changes in T cell adhesion and migration. This review highlights recent insights into our understanding of the signaling events by which the CD3/T cell receptor complex and chemokine receptors regulate integrin function and T cell migration. These studies highlight novel functions for several signaling molecules, including the tyrosine kinases Itk and ZAP-70, and the adapter protein SLAP-130/Fyb. In addition, analysis of the regulation of integrin function and chemokine-mediated migration has highlighted the critical role that spatial localization of signaling molecules plays in signal transduction, and the importance of the actin cytoskeleton in T cell function. PMID- 12637775 TI - [Serum leptin level in prepubertal children with simple obesity. Part II]. AB - The aim of the study was to determinate the serum leptin level, glucose concentration, lipids (total cholesterol, HDL-cholesterol, LDL-cholesterol, triglyceride) and thyroid hormones level (triiodothyronine, thyroid stimulating hormone and free thyroxine) in a group of prepubertal children with simple obesity. Fourfold higher leptin concentration in obese (23.3 +/- 11.8 ng/mL) in comparison to a group of slim children (6.8 +/- 2.7 ng/mL), (p<0.0001) was shown. There was no significant difference in serum glucose level (87.3 +/- 9.7 mg/dL) between both studied groups. In obese subjects we showed higher triglyceride, insignificantly lower HDL-C concentrations and invariable other lipid fractions. Mean values of triiodothyronine (T3) were 3.0 +/- 0.7 nmol/L in obese children and 2.8 +/- 0.8 nmol/L in slim children. On the contrary, mean values of thyroid stimulating hormone (TSH) were 2.5 +/- 1.7 mIU/L and 2.8 +/- 0.7 mIU/L in the group of obese and normal children respectively. Free thyroxine (FT4) concentrations in our obese children were lower (11.5 +/- 2.9 pmol/L) than in slim subjects (14.6 +/- 3.1 pmol/L) but were within the reference range. The results obtained indicate that in children with simple obesity there was no dyslipidemia connected with leptin and thyroid hormones levels. However, higher triglyceride and lower HDL-C concentrations suggest a necessity of monitoring the body mass index and lipid profile in these patients. PMID- 12637776 TI - [LCHAD (long-chain 3-hydroxyacyl-CoA dehydrogenase) deficiency as a cause of sudden death of a three months old infant]. AB - LCHAD deficiency is a rare, autosomal recessive congenial defect of fatty acid oxidation. In this disorder high mortality is observed. In some cases patients die during the first episode of this disease, without the diagnosis of LCHAD. We report the case of a three months old boy, whose sudden death was the result of LCHAD deficiency. PMID- 12637777 TI - [Cancer in children with celiac disease. Gluten-sensitive enteropathy in a boy operated for Wilms tumour]. AB - In adults untreated coeliac disease (CD) is associated with a wide variety of malignant complications. The overall mortality in CD is approximately twice that of the general population. The excess deaths are due mainly to intestinal lymphoma. In Europe, only 26 cases of CD and cancer in children have been published. There is evidence that cancer and CD in children are underreported. We report a case of Wilms tumour in a child who developed CD. As a neonate the boy had been successfully operated for congenital mesoblastic nephroma. Gluten was introduced in his diet at age of 12 months. He developed vomiting, abdominal distention ad hypertransaminosaemia. Up to the age of 16 months he had lost 1500 g and coeliac crisis was diagnosed (IgAEMA titre 1:50 and in the duodenal biopsy specimen there was crypt hyperplastic total villous atrophy). Symptoms improved rapidly when gluten-free diet was initiated, and 4 years later, clinical and serological findings were normal. Wilms tumour accounts for 6% to 7% of all childhood malignancies. There are no reports in MEDLINE about coeliac disease in patients who have been operated for the tumour. This seemed intriguing because prevalence of gluten-sensitive enteropathy calculated for Europe varies between 1/100 and 1/300. PMID- 12637779 TI - [Serum anion gap in the umbilical cord blood of healthy newborn infants]. AB - This paper presents the results of the determination of serum anion gap in arterial and venous umbilical cord blood obtained from a carefully selected group of 47 healthy term newborns. In the arterial blood, the following concentrations of particular anions were found: chloride - 102.85 +/- 4.35 mmol/l, bicarbonate - 22.26 +/- 2.67 mmol/l, protein anions - 11.89 +/- 1.00 mmol/l, and residual anions - 14.26 +/- 4.35 mmol/l, while in the venous blood these values were: chloride - 104.28 +/- 4.14 mmol/l, bicarbonate - 20.79 +/- 2.70 mmol/l, protein anions - 12.48 +/- 1.23 mmol/l, and residual anions - 13.03 +/- 3.06 mmol/l. Provided strict clinical and biochemical (acid-base balance and blood gases) selections criteria are applied, these data are likely to represent normal values for term newborns and can be the basis for comparison of umbilical serum anion gap in newborns under depressed conditions, like asphyxia. As discussed, the role of serum anion gap studies in the diagnosis of the type of metabolic acidosis, as well as in the monitoring of neonatal therapy is most important. PMID- 12637778 TI - [Somatic development assessment of children with food allergy treated with milk free diet]. AB - The aim of our study was to evaluate the influence of one year elimination diets on the somatic development and the nutritional status of children. 25 patients from the Department of Immunology of the Institute of Mother and Child aged 1-7 years with cow's milk allergy were investigated. The children were divided into two groups: 10 children received hydrolizated casein formula and 15 children received soya protein formula. The somatic development and nutritional status was assessed twice, the first time before the onset of treatment and the second time after one year of treatment. Anthropometric methods were used, body height, body mass, upper arm circumference and subscapular skinfold were measured. The data were compared with the normal Warsaw population data from 1996-99. The results showed that both the diet with hydrolizated casein formula and the soya protein resulted in normal somatic development of children. PMID- 12637780 TI - [Serum osteocalcin and bone alkaline phosphatase in healthy children in relation to age and gender]. AB - Biochemical markers of bone formation are important in the study of growth and skeletal metabolism. However, interpretation of their values for children and adolescents is difficult because they depend on many factors such as age, gender, pubertal stage, race, nutritional and health status, specificity of assays and others. Therefore, age and sex specific reference ranges for bone formation markers must be established in a defined paediatric population. The purpose of this study was the investigation of normal serum concentration of osteocalcin (OC) and bone alkaline phosphatase (BALP) in Polish children aged 2-18 years. We studied 121 healthy children (56 girls, 65 boys) divided into 3 age groups of both genders: prepubertal, pubertal and postpubertal. The level of OC was determined by N-MID Osteocalcin One Step ELISA kit (Osteometer Bio Tech, Denmark) and the activity of BALP was measured using an enzyme immunoassay Alkphase-B kit (Metra Biosystems, USA). We observed, that both formation markers showed sigmoid regression curves with increasing age. The peak values of OC and BALP occurred during puberty in girls aged 9-13 years (115.6 +/- 21.3 ng/ml; 108.8 +/- 23.6 U/L) and in boys aged 10-15 years (117.8 +/- 22.3 ng/ml; 118.4 +/- 24.5 U/L). In all children after puberty, we observed a gradual lowering of both markers. However, girls showed decreased postpubertal values of OC and BALP 2-3 years earlier than boys, indicating the earlier completion of puberty in girls. The correlation between OC and BALP was statistically significant (r=0.612; p<0.001) in tested children. The results of this study may establish the reference values for bone turnover markers in Polish healthy children, which will be useful in the diagnosis and monitoring of therapy in bone diseases. PMID- 12637781 TI - [Concentration of plasma beta-carotene in cystic fibrosis children with pancreatic insufficiency]. AB - Patients with cystic fibrosis (CF) have significantly decreased plasma concentrations of antioxidant vitamins which is considered to result both from fat malabsorption and from chronic pulmonary infection. The aim of this study was to investigate levels of vitamin A and beta-carotene in plasma of CF subjects with pancreatic exocrine insufficiency. In thirty CF patients mean plasma concentrations of vitamin A (1.17 micromol/l) was lower by 30% and that of beta carotene (1.3 micromol/l) by 60% than in healthy children. Vitamin A level was positively correlated with beta-carotene (r=0.67; p<0.001). Ratios of beta carotene to total cholesterol and cholesterol of LDL (low density lipoprotein) were 40% lower than in the control group. We conclude that in our CF patients the concentration of beta-carotene does not seem to be sufficient to inhibit lipid peroxidation especially of LDL fraction. PMID- 12637782 TI - The topical treatment of psoriasis. AB - According to the patients, improvement of efficacy, long-term safety and improvement of compliance are needed. The topical treatment has been innovated during the last decade. Most important are the introduction of two new classes of treatments: topical vitamin D(3) analogues and the retinoid tazarotene. To what extent, however, have we achieved developments which are in line with the needs as expressed by the patients? Improved efficacy has been realized by successful combinations of topical treatments. In particular, the combinations of dithranol, vitamin D(3) and tazarotene with a topical corticosteroid proved to be very effective with a reduced profile of side-effects. The efficacy of vitamin D(3) analogues and tazarotene is such that the efficacy of a potent corticosteroid (betamethasone-17-valerate) is approached; calcipotriol even showed an efficacy which is at least as good as this corticosteroid. The long-term safety of new compounds has been evaluated for at least 12 months in large studies. Remarkably for corticosteroids such information is available for only 12 weeks. However, intermittent applications of a topical corticosteroid in combination with another topical treatment provide an effective and safe long-term control of psoriasis. Compliance is a conditio sine qua non for an effective topical treatment. Important progress has been made to increase compliance. Short-contact dithranol has been popularized as an ambulatory treatment which is a highly effective approach as a care instruction programme. Formulations which are better from a cosmetical point of view have been developed for various topical treatments. Reduction of the frequency of applications proved to be possible for most treatments. Once daily applications for corticosteroids, vitamin D(3) analogues and retinoids have been developed, and intermittent applications, a few times per week, are possible for corticosteroids, which proved to be very effective with a reduced profile of side-effects, and are also developed for dithranol. PMID- 12637783 TI - Anti-acne effects of Oriental herb extracts: a novel screening method to select anti-acne agents. AB - The acne-therapeutic effects of Oriental herb extracts were investigated in terms of antichemotactic effect on polymorphonuclear leucocytes, antilipogenic actions, antibacterial activity against Propionibacterium acnes and resistance induction potency in the bacteria. The ethanol extract (0.01%) of Angelica dahurica markedly suppressed neutrophil chemotaxis, comparable to the effect of erythromycin (0.01%), whereas a strong antilipogenic effect was obtained with rhizoma coptidis (Coptis chinensis) extract (0.01%), leading to a higher efficacy than that of retinoic acid (0.01%). Interestingly, only Glycyrrhiza glabra showed a remarkable antibacterial activity against P. acnes, resulting in negligible induction of resistance, in comparison with a marked development of resistance in the bacteria treated with erythromycin. We suggest that an appropriate formulation containing A. dahurica, rhizoma coptidis and G. glabra could be helpful for the prevention and treatment of acne lesions. PMID- 12637784 TI - Formulation study and anti-inflammatory efficacy of topical semi-solids containing a nitro ester of flurbiprofen. AB - A new nitro-oxybutylester of flurbiprofen (NO-FP) is a promising anti inflammatory drug in the treatment of dermatological disorders, and the feasibility of its cutaneous administration was evaluated. Four different semi solid formulations were evaluated in order to assess the influence of the composition on the drug amount retained in the stratum corneum and epidermis (SCE). The lipophilic ointment induced the highest NO-FP amount retained in the SCE and, therefore, skin permeation enhancers (Transcutol), Lauroglycol), oleic acid and isopropyl myristate) were added to this formulation. The in vitro NO-FP amounts retained in the SCE were correlated with the solubility parameters, and a good linear correlation was found (r(2) = 0.925). The formulation of the lipophilic ointment was optimized, and the activity of this preparation was verified in methyl-nicotinate-induced contact urticaria and UV-induced erythema obtaining good results in terms of efficacy and safety. PMID- 12637785 TI - Clinical and photographic scoring of skin aging. AB - Chronic exposure to sunlight induces clinical, histological and physiological changes that are described as photoaging. To assess the resulting skin changes different clinical and photographic scores have been evaluated. Regarding different scoring systems a standardized grading system would be useful in a variety of indications, in particular to improve the quality of epidemiologic and clinical studies of photodamage. Photonumeric grading is considered superior to descriptive scales; however, the combination of both may suggest an advanced assessment of photodamage. PMID- 12637786 TI - Bifidobacterium-fermented soy milk extract stimulates hyaluronic acid production in human skin cells and hairless mouse skin. AB - We examined the effects of Bifidobasterium-fermented (BE) and nonfermented (SME) soy milk extracts on the production of hyaluronic acid (HA) in vitro and in vivo. BE, but not SME, significantly enhanced the production of HA in monolayer and organotypic cultures of human keratinocytes, in cultures of human skin fibroblasts, and in hairless mouse skin following topical application for 2 weeks. In the organotypic cultures formed by a similar structure to human epidermis, BE also extended the distribution of HA. Genistein and daidzein, known to stimulate HA production, were detected in BE at a concentration of 0.18 and 0.07 mM, respectively, but not in SME. Therefore, BE has the potential to enhance HA production in the epidermis and dermis, mainly due to genistein released from its glycoside during fermentation. BE is expected to prevent the age-dependent loss of cutaneous HA. PMID- 12637787 TI - Stereochemical considerations on concomitant allergic contact dermatitis to ester of the cis-trans isomeric compounds maleic acid and fumaric acid. AB - Allergic contact dermatitis from esters of fumaric acid or esters of maleic acid is rare. The case of a chemist with allergic reactions to esters both of fumaric acid and of maleic acid is presented. Extremely high sensitivity of the patient to diethyl fumarate was noted. The formation of identical complete antigens from esters of these two cis-trans isomeric acids may be an explanation of the patient's double allergy. This is discussed from a stereochemical point of view. These stereochemical considerations point to a general mechanism where cis-trans isomeric alpha,beta-unsaturated carbonyl compounds are converted into the same complete antigen. PMID- 12637788 TI - Predicting the dermal absorption of thalidomide and its derivatives. AB - Concern has been expressed about the ability of simple algorithms to predict skin permeability and hence skin flux. For a series of thalidomide analogues, a number of software packages have been used to predict octanol water partition coefficients. These, in conjunction with molecular weight, have then been used to calculate skin permeability coefficients. These compare favourably with experimental values. Some of the software packages also predict aqueous solubilities, which can be subsequently used to calculate maximum skin flux. The predicted and measured solubilities have been compared together with the maximum fluxes. The results show that software can be used to predict octanol water partition coefficients and aqueous solubilities (more accurately if the melting point of the compound is known) and hence to obtain very reasonable estimates of skin permeation parameters. These are useful in predicting which analogue has the most appropriate properties for dermal delivery; in the case of the thalidomide analogues, it is the methyl-substituted compound that is best. PMID- 12637789 TI - In vitro and in vivo study of 5-methoxypsoralen skin concentration after topical application. AB - The aim of this work was to study the skin distribution of 5-methoxypsoralen (5 MOP) after application of topical gels, in vitro and in vivo, in both healthy and psoriatic skin sites of 6 psoriatic patients. Drug skin distribution was determined using the thin slicing technique and subsequent HPLC analysis. In the presence of dermatological disease, i.e. psoriasis, the permeability of the tissue changed considerably, leading to an important increase in the cumulative amount of 5-MOP recovered in the skin after topical application. The amount of 5 MOP found in vitro in the human skin was intermediate between those cumulated in healthy and psoriatic skin sites during an in vivo experiment. The gel formulation is an efficacious carrier for the topical photochemotherapy of psoriasis with 5-MOP, since it allows drug penetration in psoriatic skin. PMID- 12637791 TI - Determination of IgG subclasses and avidity of antithyroid peroxidase antibodies in patients with subclinical hypothyroidism - a comparison with patients with overt hypothyroidism. AB - OBJECTIVE: To evaluate the immunoglobulin G subclasses of anti-TPO and antibody avidity in patients with subclinical hypothyroidism (sH), overt hypothyroidism (H) and a control group (C). METHODS: According to the TSH, fT4 and anti-TPO antibody levels, appraised by immunometric assays, 95 female patients were divided into three groups (sH, H and C). IgG subclass levels and avidity were measured by a homemade ELISA. Results were analyzed by nonparametric tests and Spearman's rank correlation. RESULTS: The predominant IgG subclasses detected in both case groups were IgG1 and IgG4 with a significantly higher level of IgG4 in the sH group. Consequently, the IgG1/IgG4 ratio was significantly lower in sH patients. CONCLUSION: The higher levels of IgG4 anti-TPO reduced significantly the IgG1/IgG4 ratio in sH patients. These results permit to envisage that increasing this ratio could be useful as a positive predictive factor for the development of overt disease in such patients. PMID- 12637790 TI - Endocrine and non-endocrine actions of ghrelin. AB - Ghrelin is a 28-amino-acid peptide predominantly produced by the stomach. Substantially lower amounts were detected in bowel, pancreas, kidneys, the immune system, placenta, testes, pituitary, and hypothalamus. Ghrelin displays strong growth hormone (GH)-releasing action mediated by the activation of the so-called GH secretagogue (GHS) receptor (GHS-R) type 1a. GHS-R are concentrated in the hypothalamus-pituitary unit but are also distributed in other central and peripheral tissues. Apart from the potent GH-releasing action, ghrelin has other actions including stimulation of lactotroph and corticotroph function, influence on the pituitary gonadal axis, stimulation of appetite, control of energy balance, influence on sleep and behavior, control of gastric motility and acid secretion, influence on exocrine and endocrine pancreatic function as well as on glucose metabolism, cardiovascular actions and modulation of proliferation of neoplastic cells, as well as of the immune system. The discovery of ghrelin opened many new perspectives of research in neuroendocrinology and metabolism, and even also in other fields of internal medicine as gastroenterology, immunology, oncology and cardiology. The possibility that ghrelin and/or GHS analogs, acting as either agonists or antagonists on different activities, might have clinical impact is obviously suggested and is receiving great attention. PMID- 12637793 TI - IGF-I transcript levels in whole-liver tissue, in freshly isolated hepatocytes, and in cultured hepatocytes from lean and obese Zucker rats. AB - BACKGROUND: The mechanisms underlying the maintenance of normal to high rates of linear growth and plasma insulin-like growth factor I (IGF-I) levels in spite of a low growth hormone secretion in obese children remain unknown. Among the animal models of early-onset obesity, obese Zucker (FA/FA) rats (which are homozygous for an inactivating missense mutation in the leptin receptor) are particularly appropriate, because their linear growth shows this growth hormone independence. METHODS: To study the regulation of IGF-I synthesis in this model, we have established primary cultures of hepatocytes derived from 12-week-old Zucker male obese and lean rats. The rat IGF-I gene contains six exons, and alternative splicing generates different mRNAs, one of which (called IGF-1B) has been shown to be decreased by fasting. We report steady state mRNA levels for IGF-I (all transcripts) and for IGF-IB in hepatocytes after 3 days in culture, in freshly isolated hepatocytes, and in whole-liver tissue. RT-PCRs using primers specific for IGF-I or IGF-IB were performed with two different internal competitors for quantification. RESULTS: In primary cultures of hepatocytes, the IGF-IB mRNA was increased by >50-fold (p = 0.01) in cells derived from obese animals as compared with cells from lean animals. However, these transcript levels were not significantly different when measured in freshly isolated hepatocytes or in whole liver tissue. CONCLUSIONS: Increased IGF-IB transcription could be an intrinsic characteristic of cultured hepatocytes harbouring leptin receptors that bear the FA mutation. However, the modulation of this characteristic by cell-cell interactions and by in vivo hormone and metabolic status remains to be studied. PMID- 12637792 TI - Tumor- and pregnancy-derived isoforms of human chorionic gonadotropin: biological and diagnostic relevance. AB - OBJECTIVES: Human chorionic gonadotropin (hCG) and hCG variants are of high clinical importance for the diagnosis of pregnancy, monitoring of abnormal and ectopic pregnancies, testing for Down's syndrome or monitoring therapy of hCG secreting malignancies. In serum and urine, hCG appears in microheterogeneous isoforms with respect to protein backbone structure and the extent of glycosylation. The present study reports on the identification, immunological characterization, biological activity of glycosylation isoforms of pregnancy (preg) and tumor-derived (tu) hCG, and the impact of glycosylation on diagnostic immunoassays. METHODS: Twenty-two urinary preg- and tu-hCG isoforms were separated by preparative isoelectrofocusing (hCG-pI variants) and characterized by Western blot. Number, topography and accessibility pattern of epitopes on their surface was evaluated by two-site radioimmunoassays using 14 different monoclonal antibodies (mabs). Binding of hCG isoforms to four different LH/CG receptors was investigated in radioreceptor assays, and their biological activity determined by measuring cAMP elevation. RESULTS: All 22 hCG glycosylation variants appeared immunologically intact: each isoform, even when highly acidic, expressed all 14 surface epitopes which were arranged in a topographical manner indistinguishable from crude hCG. hCG isoforms were able to bind to four different receptor variants, with slightly varying affinities, but orientations indistinguishable from each other as shown by identical epitope accessibility patterns. Each of the hCG-pI variants was able to activate the LH/CG-Rs, but with varying reactivities. CONCLUSIONS: We conclude that in contrast to deglycosylated hCG, all hCG glycosylation isoforms investigated act as receptor agonists. Moreover, there is no overspecificity of mabs to certain hCG isoforms due to carbohydrate variability that exclude others from diagnostic measurement. PMID- 12637794 TI - Effect of natriuretic peptides on in vitro stimulated adrenocorticotropic hormone release and pro-opiomelanocortin mRNA expression by the fetal rat pituitary gland in late gestation. AB - OBJECTIVE AND METHODS: We investigated the effects of individual natriuretic peptides (atrial natriuretic peptide, ANP; brain natriuretic peptide, BNP, and C type natriuretic peptide, CNP) on rat corticotropin-releasing factor stimulated adrenocorticotropic hormone (ACTH) secretion by the pituitary gland of 21-day-old rat fetuses in vitro and on pro-opiomelanocortin gene expression using in situ hybridization. RESULTS: Graded concentrations of ANP, BNP, or CNP (10(-10), 10( 9), and 10(-8) mol/l) induced a log dose dependent inhibition of ACTH secretion induced by rat corticotropin-releasing factor (10(-10) mol/l). These natriuretic peptides showed equipotent effects on a molar basis. Moreover, ANP, BNP, or CNP at 10(-10) mol/l reduced significantly the pituitary pro-opiomelanocortin mRNA expression. In addition, the immunoreactive ANP, BNP, and CNP cells were localized in the anterior lobe, but not in the intermediate lobe of the fetal pituitary gland. CONCLUSIONS: These data suggest that the fetal pituitary gland may be both a source and a target for natriuretic peptides that might control ACTH synthesis and release via an endocrine and/or paracrine mechanism. The natriuretic peptides could participate, as well as glucocorticoids, in the control of the corticotropin-stimulating activity of the fetal rat in late gestation. PMID- 12637795 TI - Inhibition of growth hormone excess reduces insulin resistance and ovarian dysfunction in a lean case of polycystic ovary syndrome with a growth-hormone producing pituitary adenoma. AB - A 23-year-old female with polycystic ovary syndrome (PCOS) and a growth-hormone (GH)-producing pituitary adenoma is described. A reduction in the elevated GH levels to normal levels following the administration of dopaminergic agents decreased plasma insulin-like growth factor (IGF)-1 and ovarian dysfunction. Menstrual cycles were therefore restored and the number of ovarian cysts reduced, suggesting that insulin and/or IGF-1, stimulators of theca cell proliferation, may be pathogenetic factors in PCOS. PMID- 12637796 TI - Influence of growth hormone therapy on pituitary and lumbar spine astrocytomas. A clinical observation. AB - We report on a male patient with an inactive pituitary and lumbar astrocytoma. He presented with panhypopituitarism at the age of 16 and received growth hormone replacement therapy for one year. During this period the pituitary mass increased substantially and showed subsequently a consistent shrinkage once treatment was stopped. The lumbar lesion stayed stable for the whole period of observation. The presumable rGH related change of the pituitary astrocytoma underlines the importance of careful follow-up of children with growth hormone deficiency secondary to treatment for brain tumors. Substitution therapy with rGH requires further and longer studies before it can be recommended with absolute reassurance in children with subtotal resection of a pituitary astrocytoma. PMID- 12637797 TI - Incisional hernias. I. Related risk factors. AB - Incisional hernias represent one of the most frequent complications of abdominal surgery. The incidence is probably underestimated. The pathogenesis is complex and not fully understood, implying patient-related factors (i.e., collagen biochemistry, obesity, age) as well as technical factors, including, among others, wound infection, suture material, and types of incisions and closures. In this paper, the first of two, the authors review the literature emphasizing the current knowledge concerning the pathogenesis of incisional hernias. The second article is focused on the treatment. PMID- 12637798 TI - Giant splenic artery aneurysm associated with chronic pancreatitis. PMID- 12637799 TI - Simultaneous laparoscopic cholecystectomy and percutaneous papillary balloon dilatation for cholecystocholedocholithiasis. AB - AIM: We evaluated the effectiveness of simultaneous laparoscopic cholecystectomy (LC) and percutaneous papillary balloon dilatation (PPBD) under general anesthesia in the treatment of cholecystocholedocholithiasis in elderly patients in poor general condition. PATIENTS AND METHODS: We treated 17 patients (group 1) with LC and PPBD as separate procedures and 21 patients (group 2) with simultaneous LC and PPBD under general anesthesia. We compared the success rates, complications, durations of percutaneous transhepatic biliary drainage (PTBD) and hospital stays of the 2 groups. RESULTS: The stones were successfully pushed out into the duodenum in all patients in both groups. All patients in group 1 complained of transient pain caused by balloon dilatation of the papilla. In group 2, PPBD was performed comfortably in all patients because of the general anesthesia. The mean postoperative stay of patients in both groups was 9 days. The overall duration of the hospital stay and the duration of PTBD were 28 +/- 5 and 25 +/- 9 days, respectively, in group 1, and 18 +/- 7 and 15 +/- 8 days in group 2 (significant difference: p < 0.01 and p < 0.01, respectively). There were no deaths or major complications, although transient hyperamylasemia occurred in both groups. CONCLUSIONS: Cholecystocholedocholithiasis can be treated by simultaneous LC and PPBD under general anesthesia without laparotomy, sphincterotomy, choledochotomy or the discomfort arising from papillary dilatation. PMID- 12637800 TI - Plasma lipids and lipoprotein changes after biliopancreatic diversion for morbid obesity. AB - BACKGROUND: The reduction in plasma cholesterol with increase in large and lower dense LDL (pattern A) obtained by statins is usually associated with a prompt reduction in cardiovascular risk, but after bariatric surgery for morbid obesity a delay of some years is observed. No data regarding LDL pattern are available in obese subjects after biliopancreatic surgery. OBJECTIVE: To evaluate the modifications in LDL composition and LDL density after biliopancreatic surgery. SUBJECTS: 29 patients (17 type 2 diabetics (type 2) and 12 non-diabetics (ND)) with BMI <35, who failed previous attempts to decrease weight by diet, were studied before and 6 months after biliopancreatic diversion for morbid obesity. MEASUREMENTS: In all subjects, besides fasting circulating lipids, glucose and insulin, LDL and VLDL composition were determined and LDL density was evaluated as well. RESULTS: After surgery we observed a significant reduction of all circulating lipids, including apolipoprotein (Apo) B. The decrease was more marked for total cholesterol (-41%) than for triglycerides (-28%), without a significant difference between type 2 and ND. After surgery, LDL presented a marked decrease in the percentage of cholesterol (from 36 to 32%) with a marked increase in the percentage of triglycerides (from 13 to 18%), without appreciable modification of ApoB. After surgery, 1 patient changed from pattern B to A, while 2 patients previously pattern A became pattern B. Also a decrease in HDL and ApoAI was evident in all the subjects with an increase in the VLDL-1. CONCLUSIONS: Our data indicate that after biliopancreatic diversion, the plasma lipid profile improves along with improvement of plasma glucose and insulin sensitivity, but the LDLs become richer in triglycerides. It is possible that the greater atherogenicity of these LDLs is compensated by an improvement in the general metabolic condition. PMID- 12637801 TI - Effects of PEEP on residual vascularization in oesophageal substitution gastroplasty by surface oximetry-capnometry and photoplethysmography: an experimental study. AB - OBJECTIVE: To evaluate the effects of positive end-expiratory pressure (PEEP) on residual vascularization in gastric tubes for oesophageal replacement. DESIGN: Experimental open study. MATERIALS: Eleven mongrel dogs. METHODS: Intestinal parietal blood flow was evaluated by photoplethysmography (PPG) and measurement of surface oxygen (PsO(2)) and carbon dioxide (PsCO(2)) tensions under basal conditions. After Akiyama's tubular gastroplasty, three levels of PEEP were administered. At each level, fluids were infused to counter the drop in cardiac output. PPG, surface gas tensions, arterial pressure, cardiac output and arterial blood gas tensions were monitored. Control sections of the bowel were also monitored by PPG. RESULTS: Cardiac output dropped for each level of PEEP and returned to basal levels on volume restabilization and on removal of PEEP. Central venous pressure and pulmonary arterial and capillary pressures increased for each level of PEEP and only returned to basal levels on removal of PEEP. PsO(2) values dropped for each level of PEEP and returned to basal levels on volume restabilization and on removal of PEEP. PsCO(2) levels rose, and PPG wave amplitude dropped, for each level of PEEP; these two variables only returned to basal levels on removal of PEEP. PPG values for the control sections reflected those of the anastomotic area. CONCLUSIONS: PEEP affects surface oxygen values at the level of the gastroplasty by means of its effect on cardiac output. PEEP also creates a venous return compromise and PPG wave amplitude and surface carbon dioxide values are related to this compromise. All three variables could be significant in anastomotic wound healing. PMID- 12637802 TI - Magnetic resonance cholangiopancreaticography as a diagnostic tool for common bile duct stones: a comparison with ERCP and clinical follow-up. AB - BACKGROUND/AIMS: The diagnostic potential of magnetic resonance cholangiopancreaticography (MRCP) has improved as a result of evolving technique. MRCP has the advantage of negligible morbidity and mortality in contrast to endoscopic retrograde cholangiopancreatography (ERCP). This study was performed to evaluate MRCP as a replacement for diagnostic ERCP for the suspicion of common bile duct (CBD) stones. METHODS: From 1998 to 2001, MRCP was performed in 202 patients with a suspicion of CBD stones based on medical history (MH), cholestatic liver function tests (CL), both MH and CL or other reasons. ERCP was performed in all patients where MRCP indicated the presence of CBD stones and in those patients with a persistent strong clinical suspicion for CBD stones despite a negative MRCP. RESULTS: In 25 patients, MRCP suggested CBD stones which were proven with ERCP in 24 patients. Despite a negative MRCP, 27 patients had a subsequent ERCP. None of these patients appeared to have CBD stones. In this group, MRCP resulted in 100% sensitivity and 96% specificity in detecting CBD stones. Follow-up of all patients revealed 5 more patients with persistent clinical suspicion or cholestatic liver function values. Assuming CBD stones in these patients, MRCP had a sensitivity of 83 % and a specificity of 99% for this diagnosis. CONCLUSION: In the case of CBD stone suspicion, MRCP should be the diagnostic procedure of choice. PMID- 12637803 TI - Primary splenic hydatidosis. AB - BACKGROUND: A hydatid cyst is still a serious health problem in endemic areas. Invasion of the spleen is rare in hydatid disease. In the medical literature, reports about isolated splenic hydatidoses are quite rare. Also there is not a consensus about the way of treatment. METHODS: Between 1978 and 2000, in our clinic approximately 900 patients were operated on for an abdominal hydatidosis; 850 of these patients were treated for a hepatic hydatidosis and 14 patients for an isolated splenic hydatidosis. Six (42.8%) of the latter patients were male and 8 (57.1%) were female. The mean age of the patients was 47.14 +/- 4.9 (range 17 72) years. Ten patients (71.4%) presented with a painful mass in the left upper quadrant of the abdomen, and the other 4 patients (28.5%) were asymptomatic and were diagnosed incidentally. RESULTS: All of the patients underwent elective splenectomy. There was no mortality, but complications occurred in 4 (28.5%) patients. The period of hospitalization ranged from 7 to 17 days with a mean of 9.8 days. 1 patient died from an acute myocardial infarction during the 2nd postoperative year, and 1 patient died as a consequence of a traffic accident during the 5th postoperative year. Nine patients, after follow-up periods of between 2 and 14 years, are living free from disease. No recurrence occurred in any of them. In the remaining 3 patients, long-term follow-up could not be maintained. CONCLUSIONS: A hydatid cyst must be included in the differential diagnosis of cystic lesions of the spleen. A splenic hydatid cyst should be treated surgically due to the high risk of a rupture, and the ideal procedure in adulthood is standard splenectomy. PMID- 12637805 TI - Video-assisted gasless transanal endoscopic microsurgery: a review of 217 cases of rectal tumors over the past 10 years. AB - BACKGROUND: Transanal endoscopic microsurgery (TEM) has been suggested as a minimally invasive procedure of low morbidity for rectal tumor excision. However, the complicated setup of TEM instruments and the expensive cost are disadvantages to wider acceptance. The aims of this study are to introduce a new modality of modified video-assisted gasless TEM (V-TEM) using a video system and no CO(2) insufflation of the rectum, and discuss our experience during past 10 years. METHODS: 217 patients, with a mean follow-up of 61 months, underwent V-TEM for adenoma (n = 102), Tis (n = 83), T1 (n = 28) and T2 (n = 4) rectal tumors, located 3-20 cm from the dentate line. RESULTS: The mean size of the tumor was 39 mm, and the mean duration of the operation was 63 min including setup time, and the mean duration of hospital stay was 5.8 days. Seven (3.2%) patients underwent conversion to radical surgery owing to T1 with massive invasion or T2 tumors histopathologically. Two (0.9%) patients had recurrent disease which was managed by repeat V-TEM. The postoperative course in all patients was free from any significant complications. Transient fecal soiling was present in 12 (5.5%) patients. The cost of V-TEM was decreased compared to that of conventional TEM (USD 40000). CONCLUSION: V-TEM was a cheap, safe, simple and minimally invasive procedure for benign and early cancer in the proximal rectum. PMID- 12637804 TI - Isolated gastric varices: prevalence, clinical relevance and natural history. AB - BACKGROUND: Isolated ectopic gastric varices (IGV2) are present either in the body or antrum of the stomach or upper duodenum. The prevalence, natural history and clinical significance of these varices has not been adequately described. MATERIALS AND METHODS: Consecutive patients with portal hypertension, prospectively studied and diagnosed to have IGV2, were assessed for their time of appearance - primary (at first presentation) or secondary (after obliteration of oesophageal varices), association with other varices, portal hypertensive gastropathy and any overt bleeding. RESULTS: Fifty-three of the 1128 (4.7%) patients had IGV2. The IGV2 were commonly seen in the antrum (53%), duodenum (32%), or at both sites (11%) and rarely in body and fundus (4%). IGV2 were predominantly (84.9%) secondary in origin, developing after oesophageal variceal obliteration. The median time for emergence of secondary IGV2 was 8.2 months for patients with cirrhosis, 12.8 months for non-cirrhotic portal fibrosis and 10.8 months for extra-hepatic portal vein obstruction. Eight (15%) patients had primary IGV2, 6 of them had underlying portal vein obstruction. Portal gastropathy (p < 0.05) and UGI bleeding were more common in the secondary than in primary IGV2. Bleeding due to IGV2 was seen only in 3 (5.7%) patients during a mean follow-up of 36.3 +/- 12.1 months, and could be successfully managed with endoscopic ligation or obliteration. CONCLUSIONS: Isolated ectopic gastric varices are not uncommon and generally develop following obliteration of main variceal columns. They rarely bleed and often can be managed with endoscopic interventions. PMID- 12637806 TI - Fistula involving portal vein and duodenum at the site of a duodenal ulcer in a patient after previous extrahepatic bile duct resection and brachytherapy. AB - Fistulas involving the portal venous system and gastrointestinal (GI) tract are rare. However, they can cause life-threatening GI hemorrhage. A case of a fistula between the main portal vein and the posterior wall of the duodenal bulb at the site of a duodenal ulcer in a patient who had previously undergone an extrahepatic bile duct resection and brachytherapy for mucinous cystadenocarcinoma is reported. Consideration should be given to this entity in the differential diagnosis of GI hemorrhage in patients with a history of previous major biliary surgery. PMID- 12637807 TI - Retroperitoneal bronchogenic cyst as a differential diagnosis of pancreatic mucinous cystic tumor. AB - Cystic tumors of the pancreas where a pseudocyst has not been able to be excluded has been considered potentially proliferative and pre-malignant or malignant and thus aggressive surgical approach has been advocated. Retroperitoneal cystic tumors are rare and among these bronchogenic cysts are extremely infrequent. The present paper describes a case of bronchogenic cyst in association with the pancreas in which diagnostic work-up was not able to exclude a proliferative pancreatic cystic tumor. PMID- 12637808 TI - Enterolith ileus as a complication of jejunal diverticulosis: two case reports and a review of the literature. AB - BACKGROUND: In a period of 5 years, 2 patients with enterolith ileus, caused by jejunal diverticulosis, were treated in our hospital. In order to learn more about treatment options, the literature was reviewed. METHODS: The case history of the 2 patients is described. Relevant articles were identified using Medline and PubMed. Data regarding patient gender, age, operative findings, therapeutic measures and outcome were collected. RESULTS: Including patients reported in the literature, 34 cases of intestinal obstruction due to enteroliths expelled from jejunal diverticula were identified. A distinction is made between complicated and uncomplicated enterolith ileus. If there are signs of bowel ischemia, other (unborn) enteroliths, inflammation of the bowel or if there are signs of a (sealed) perforation, the case is considered a complicated enterolith ileus. If none of these signs are present, uncomplicated enterolith ileus is present. In uncomplicated enterolith ileus (21 patients), more often milking and crushing or enterotomy was performed. In complicated enterolith ileus (13 patients), more often a segmental resection of the involved jejunum was performed (p < 0.01). CONCLUSION: Small bowel obstruction due to enteroliths expelled from jejunal diverticula is a rare condition. Relevant literature is only available in the form of case reports. On the basis of the presented patients and patients reported in the literature, a justifiable therapeutic strategy is presented. The least invasive step in the therapeutic approach is to crush and milk the obstructing enterolith down to the colon. Laparoscopic crushing and milking of the enterolith is described. If this fails an enterotomy could be tried, if possible proximal or distal from the obstruction site, in order to make an incision in a less edematous area. If the first two strategies fail, or if complicated enterolith ileus is present, resection of the involved jejunal segment could be considered. PMID- 12637809 TI - Management of hepatobiliary cystadenocarcinoma. AB - Primary biliary cystadenocarcinomas are rare malignant neoplasms of the liver. We report a case that was operated with a preoperative diagnosis of hydatid cyst of the liver. The patient underwent a wide surgical excision of the cyst. Histopathological examination revealed a hepatobiliary cystadenocarcinoma. The patient remains asymptomatic at 2 years following surgery. The importance of differentiating these rare tumors from hepatic hydatid cysts, especially in areas where the disease is endemic, is highlighted in this case report. PMID- 12637810 TI - Carcinoma of pelvic pouch following restorative proctocolectomy: report of a case and review of the literature. AB - One of the possible long-term complications following an ileoanal pouch procedure is the development of malignancy in the mucosa of the rectal stump. Only 10 such cases have been reported so far. We report the case of a 23-year-old male who had ulcerative colitis with high-grade dysplasia and underwent an ileoanal pouch procedure. He was found to have a malignancy in the rectal stump 5 years after surgery. The malignancy presented with symptoms of refractory pouchitis. Pouch excision was performed. The risk of developing malignancy in the rectal mucosa after an ileoanal pouch procedure mandates regular follow-up in these patients, with a high index of suspicion especially in patients with delayed onset pouchitis. PMID- 12637811 TI - Pancreatic arteriovenous malformation. AB - We report a case of a 57-year-old male with a pancreatic arteriovenous malformation (AVM). The diagnosis was made by CT scan and angiography. In the reported case, bleeding from AVM into the biliary tract caused haemobilia and bleeding into the pancreatic head caused abdominal pain. Pancreatic AVM has been only rarely reported as a cause of haemobilia. PMID- 12637812 TI - A case of large mucinous adenocarcinoma arising in a long-standing fistula-in ano. AB - BACKGROUND: Chronic anal fistulas are not rare conditions, however, the development of a carcinoma in a long-standing fistula-in-ano is rare. METHODS: The case of a 77-year-old male with a large perianal mucinous adenocarcinoma arising in a long-standing fistula-in-ano is presented. RESULTS: Perianal biopsy revealed mucinous adenocarcinoma. Abdominal CT, double contrast barium examination and flexible sigmoidoscopy revealed no other tumoral lesion in the colon and rectum. CONCLUSION: The patient underwent abdominoperineal resection including wide tumor excision on the gluteal region. The final reconstruction was performed by bilateral gracilis musculocutaneous flaps. Due to clinical and histopathological evidence it was thought that a curative resection had been performed. To date he is clinically disease free. PMID- 12637813 TI - A true splenic cyst producing carbohydrate antigen 19-9 and cancer antigens 50 and 125, but not interleukin 10. AB - A giant cystic lesion of the left upper abdomen associated with a smaller ovarian cyst in a young female patient is reported. Laboratory data revealed elevated serum levels of carbohydrate antigen 19-9 (CA 19-9), carcino-embryonic antigen (CEA), cancer antigens 50 and 125, and tissue polypeptide antigen. In contrast, the serum levels of interleukin 10, a cytokine involved in modulating immune responses and produced by many cancer histotypes, were normal. Since ovarian cancer or cystic adenocarcinoma of the tail of the pancreas were not ruled out, the patient underwent laparotomy. After splenectomy and ovariectomy, the tumour marker serum levels normalized. Histology and immunohistochemical analysis revealed a true splenic cyst with the inner epithelium strongly positive for CA 19-9 and CEA and high levels of cancer antigens in the fluid. The ovarian lesion was a serous cystadenoma. The inner epithelium showed no immunoreactivity for tumour markers which were not measurable in the fluid. True cysts of the spleen are rare; in a few cases, high serum levels of CA 19-9 and CEA have been reported. In such instances, cyst resection or splenectomy is indicated to rule out malignant lesions and to remove the cancer antigen producing epithelium. The reported case shows that the epithelium lining true splenic cysts may produce, besides CA 19-9 and CEA, other tumour markers, in particular cancer antigens 50 and 125. In addition, normal serum values of interleukin 10 correctly predicted the benign nature of the lesion. PMID- 12637814 TI - Open and laparoscopic treatment of nonparasitic splenic cysts. AB - BACKGROUND: Nonparasitic splenic cysts are rare. Therefore, there is no 'evidence based' information regarding their optimal surgical management. In the last years the laparoscopic approach has gained increasing acceptance in splenic surgery. The aim of this study is to present our experience with the laparoscopic management of splenic cysts. METHODS: The medical records of 7 patients with splenic cysts were reviewed retrospectively. RESULTS: One patient had an open partial splenic resection. Five patients, 3 of them with a posttraumatic and 2 with an epidermoid splenic cyst, underwent laparoscopic unroofing of the cyst. In 4 of these cases the postoperative course was uneventful, whereas in 1 case the patient developed a cyst relapse soon postoperatively. Later on this patient successfully underwent an open partial splenic resection. The 7th patient had an explorative laparoscopy. The cyst was located intrasplenically, entirely covered with unaffected splenic parenchyma, and reached the splenic hilus. Therefore, a conversion to open partial splenectomy was performed. CONCLUSION: Open partial splenectomy and laparoscopic cyst wall unroofing are both effective tools in the management of splenic nonparasitic cysts. Surgeons must master both techniques as nowadays spleen-preserving techniques should be attempted in every case of splenic nonparasitic cyst. PMID- 12637815 TI - Recent studies show nursing's importance and integrity. PMID- 12637816 TI - HHS and ADA warn Americans of "prediabetes". Individuals encouraged to take healthy steps to reduce risks. PMID- 12637817 TI - End-stage dementia: hospice offers expert interdisciplinary care for end-stage dementia. PMID- 12637818 TI - What's new in the 2003 Joint Commission? Home care and hospice standards: part 2. PMID- 12637819 TI - Phlebotomy tools of the trade: part 3: Alternative sites for drawing blood. PMID- 12637820 TI - CMS clarifies posthospital referrals. PMID- 12637821 TI - Assisting patients using positive airway pressure therapy. AB - The number of patients with sleep apnea treated by home care nurses is increasing. The most common treatment is with at-home positive airway pressure (PAP). This article presents an overview of PAP, including its purpose, correct application, and hints for assisting patients when they experience problems. Additionally, resources for further information on this therapy for sleep apnea are provided. PMID- 12637822 TI - Central venous access device occlusions: part 2: nonthrombotic causes and treatment. AB - Home care nurses commonly care for patients with central venous access devices (CVAD) for administration of a variety of infusion therapies. Catheter occlusion is a potential complication. In the February 2002 issue of the etiology and treatment of thrombotic occlusions were described. This month, the focus is on nonthrombotic causes of CVAD occlusion. PMID- 12637823 TI - Helping Hispanic/Latino home health patients manage their pain. AB - The research focusing on pain in Hispanic/Latino populations suggests that their cultural values and beliefs of stoicism, fatalism, the importance of family, spirituality, and folk healing have an impact on their pain experience. Based on research findings this article suggests strategies nurses can use to assess and suggest pain management interventions for patients of Hispanic/Latino culture. PMID- 12637824 TI - Decreasing back stress in home care. AB - Although back injuries are a significant problem in home care, limited information exists regarding which tasks are stressful and how stress can be reduced. This study describes home health aides' perceived stressfulness to the back and provides ideas all home care workers can use to decrease back stress. PMID- 12637825 TI - Medicare survey shifts focus to outcomes. PMID- 12637827 TI - Caregiver Strain Index (CSI). PMID- 12637826 TI - Hospice and palliative care specialty certification for nursing assistants. PMID- 12637828 TI - "I'm so proud of us!". PMID- 12637830 TI - Is esophoria a factor in slowing of myopia by progressive lenses? PMID- 12637831 TI - Corneal response to short-term orthokeratology lens wear. AB - PURPOSE: This study investigated short-term corneal changes induced by reverse geometry lenses worn for orthokeratology. METHODS: Nine young adult subjects wore reverse-geometry rigid gas-permeable lenses (BE; UltraVision Contact Lenses, Brisbane, Australia) in one eye only for 10, 30, and 60 min in the open eye and 8 h in the closed eye. The fellow eye acted as a non-lens-wearing control. Corneal topographic changes were monitored using the Medmont E-300 corneal topographer and keratometry. Changes in uncorrected logarithm of the minimum angle of resolution (log MAR) visual acuity were also recorded. Data were analyzed by analysis of variance and t-tests. RESULTS: Significant central corneal flattening (-0.61 +/- 0.35 D; p = 0.014) and the formation of a defined "treatment zone" (diameter, 3.86 +/- 0.88 mm) were found after 10 min of open-eye lens wear, which progressed with increasing periods of lens wear. Significant improvement in unaided logMAR (-0.16 +/- 0.18; p = 0.005) was also apparent after 10 min and showed further improvement with longer periods of lens wear. Corneal asphericity showed a trend toward corneal sphericalization, which reached statistical significance after 8 h of lens wear. There was no significant change in corneal toricity. CONCLUSIONS: The cornea responds rapidly to the application of reverse geometry lenses for orthokeratology, with significant central corneal flattening and improvement in visual acuity after just 10 min of lens wear. This suggests that the corneal epithelium is able to be molded or redistributed very rapidly in response to the tear film forces generated behind reverse-geometry lenses. PMID- 12637832 TI - Driving habits and health-related quality of life in patients with age-related maculopathy. AB - PURPOSE: To characterize the driving habits of persons with age-related maculopathy who present to a low-vision rehabilitation clinic and to examine how driving status relates to vision-specific health-related quality of life. METHODS: The Driving Habits Questionnaire, the National Eye Institute Vision Function Questionnaire-25, and the Life Space Questionnaire were administered via telephone interview to 126 patients with age-related maculopathy who presented to a low-vision clinic during the previous year and were either past or current drivers. RESULTS: Twenty-four percent of the sample reported being a current driver. Compared with those who stopped driving, current drivers were more likely to be male, younger, have better visual acuity and higher National Eye Institute Vision Function Questionnaire-25 scores. Drivers reported driving an average of 4 days and 10 miles per week. Over 50% of drivers reported that because of their vision, they had difficulty with or did not drive at all in rain, at night, on freeways or interstate highways, in heavy traffic areas, or during rush hour. Drivers and nondrivers did not differ in their life space, the spatial extent of their excursions into their environment. CONCLUSION: Some individuals who present to a low-vision clinic with age-related maculopathy do drive, although their driving exposure is low and they report avoiding challenging on-road situations. Driving status in age-related maculopathy appears to be related to better eye visual acuity and vision-specific health-related quality of life. PMID- 12637833 TI - Nearpoint of convergence: test procedure, target selection, and normative data. AB - BACKGROUND: The purpose of this study was to help determine the most appropriate target to be used for the assessment of the nearpoint of convergence, normative data for the break and recovery in adults, and the diagnostic value of the red glass modification and repetition of the nearpoint of convergence. METHODS: A total of 175 subjects with normal binocular vision and 38 subjects with convergence insufficiency were evaluated. The nearpoint of convergence was measured three ways, with an accommodative target, a penlight, and a penlight with red and green glasses. The nearpoint of convergence was also measured using a penlight for 10 repetitions. RESULTS: Results suggest a clinical cutoff value of 5 cm for the nearpoint of convergence break and 7 cm for the nearpoint of convergence recovery with either an accommodative target or a penlight with red and green glasses. CONCLUSION: This study establishes normative data for the nearpoint of convergence break and recovery in the adult population and supports the value of various test modifications when other testing is equivocal. PMID- 12637834 TI - Ocular component data in schoolchildren as a function of age and gender. AB - PURPOSE: To describe the refractive error and ocular components of a large group of school-aged children as a function of age and gender. METHODS: In this report, we describe the refractive error and ocular components of 2583 school-aged children (49.3% girls, overall mean [+/-SD] age 10.0 +/- 2.3). Measurement methods included cycloplegic autorefraction, autokeratometry, videophakometry, and A-scan ultrasonography. For statistical comparisons across gender and age, a critical point of alpha = 0.005 was used to assess significance because of the large sample size and the large number of comparisons made. RESULTS: Of these 2583 children, 10.1% were myopic (-0.75 D or more myopia in both meridians), and 8.6% were hyperopic (+1.25 D or more hyperopia in both meridians). As would be expected, there was a significant effect of age on refractive error (spherical equivalent, p < 0.0001), toward less hyperopia/more myopia. There was no significant difference in the average refractive error between girls and boys (p = 0.0192). Girls had steeper corneas than boys (0.74 D steeper in the vertical meridian and 0.63 D steeper in the horizontal meridian, p < 0.0001). There were no significant differences in corneal power with age (p = 0.16). Both older age and male gender were significantly associated with deeper anterior chambers (p < 0.0001 for both). The crystalline lens showed significant thinning with age (p < 0.0001), however, there was no significant difference in the lens thickness between girls and boys (p = 0.66). Both Gullstrand lens power and calculated lens power showed significant effects of age and gender (p < 0.0001 for both). Girls, on average, had Gullstrand lens powers that were 0.28 D steeper and calculated lens powers that were 0.80 D more powerful than boys. Axial length also showed significant effects of age and gender (p < 0.0001 for both). Girls' eyes were, on average, 0.32 mm shorter than those of boys. CONCLUSIONS: These cross-sectional data show a general pattern of ocular growth, no change in corneal power, and crystalline lens thinning and flattening between the ages of 6 and 14 years. Girls tended to have steeper corneas, stronger crystalline lenses, and shorter eyes compared with boys. PMID- 12637835 TI - The effect of illumination-microscope angle on slit lamp estimate of the anterior chamber depth. AB - PURPOSE: The anterior chamber depth can be estimated by the slitlamp without additional attachments. The method involves increasing the length of a horizontal slit focused on the cornea until the corneal and iris/lens images appear to just touch. For a slitlamp angle of 60 degrees, multiplying the just-touching slit length (JTSL) by 1.4 gives an estimate of the anterior chamber depth as measured by ultrasonography. The purpose of this study was to determine whether the slitlamp angle affects the accuracy of the procedure. METHODS: The anterior chamber depth of 63 adult male and female subjects aged between 19 and 44 years was measured by A-scan ultrasonography and estimated by the slitlamp technique. The angle between the illumination and microscope systems was varied at 10 degrees intervals from 30 degrees to 70 degrees. RESULTS: The average ultrasonographic anterior chamber depth for all subjects was 3.23 +/- 0.38 mm. The average JTSL was as follows: 30 degrees = 1.5 mm, 40 degrees = 1.7 mm, 50 degrees = 2.1 mm, 60 degrees = 2.3 mm, and 70 degrees = 2.5 mm. The conversion ratio between the ultrasonographic anterior chamber depth and the average JTSL was as follows: 30 degrees = 2.2, 40 degrees = 1.9, 50 degrees = 1.5, 60 degrees = 1.4, and 70 degrees = 1.3. The ultrasonic anterior chamber depth was best estimated at 60 degrees. The 95% limits of agreement between the estimated and measured anterior chamber depth were as follows: 30 degrees = -0.76 to +0.84 mm, 40 degrees = -0.75 to +0.63 mm, 50 degrees = -0.36 to +0.46 mm, 60 degrees = 0.26 to +0.28 mm, and 70 degrees = -0.45 to +0.41 mm. CONCLUSIONS: These results experimentally confirm Smith's suggestion that the optimum slitlamp angle at which to accurately estimate the anterior chamber depth is 60 degrees. At this angle, estimates of the anterior chamber depth are minimally deviated from the gold standard (A-scan ultrasonography). At a slitlamp angle of 60 degrees, the JTSL can be converted to an estimate of the ultrasonic anterior chamber depth by multiplying it by 1.40. PMID- 12637836 TI - Repeatability and validity of the PowerRefractor and the Nidek AR600-A in an adult population with healthy eyes. AB - We assessed the repeatability and validity of the PowerRefractor and the Nidek AR 600A autorefractor. This is the first independent study conducted on adults to evaluate the performance of these instruments in a laboratory setting. Fifty subjects (23 males and 27 females) aged 16 to 61 years (mean, 37 +/- 12) participated in the study. The validity of the PowerRefractor and the Nidek autorefractor readings were determined by comparing them to subjective refraction. Measurements of refractive error were obtained from the two instruments on two separate occasions to assess their repeatability. The measured refractive error was converted into a dioptric power matrix for data analysis. No significant difference was found between the measurements obtained with the two instruments and the subjective refraction. The estimate of refractive error given by the two instruments was also found to be repeatable. In addition to measuring the refractive error, the PowerRefractor also offers the facility to measure eye position, pupil size, and dynamics of accommodation. We suggest some improvements to the PowerRefractor measurement technique to standardize its clinical use and to improve accuracy. PMID- 12637837 TI - The construction of a model eye for investigation of laser-tissue interactions in scanning laser ophthalmoscopy. AB - PURPOSE: To develop a model eye to study laser-tissue interactions during retinal imaging with scanning ophthalmoscopy. METHODS: A model eye was designed to match the optical properties of the human eye based on the Bennett and Rabbetts schematic eye. RESULTS: Alterations in axial length resulted in changes in refractive error similar to those in the human eye (3.70 D for 1 mm in axial length). Perfusion-fixed retinal tissue could be successfully imaged using the Heidelberg Retina Tomograph and Optical Coherence Tomograph to provide images that are similar in quality to those obtained. CONCLUSIONS: The model eye should be a valuable tool for investigating laser-tissue interactions during scanning laser ophthalmoscopy and the derivation of digital retinal and tomographic images. This model should also enable a determination of the accuracy with which digital imaging techniques, such as the optical coherence tomograph and Heidelberg Retina Tomograph, measure retinal structure. PMID- 12637838 TI - A cost-effectiveness model of screening strategies for amblyopia and risk factors and its application in a german setting. AB - PURPOSE: To develop a general setting-independent decision-analytical model that determines the costs, effectiveness, and cost-effectiveness of four screening strategies to detect amblyopia or amblyogenic factors in pre-school children and to apply the model in a German setting. METHODS: The general setting-independent decision-analytical model was developed from the perspective of society and the statutory health insurance was developed. Outcomes were the total number of newly detected true positive cases of amblyopia and the costs per newly detected true positive case of amblyopia. Strategies were screening of high-risk children up to the age of 1 year (ophthalmologists), screening of all children up to the age of 1 year (ophthalmologists), screening of all children aged 3 to 4 years (pediatricians or general practitioners), and screening of children aged 3 to 4 years visiting kindergarten (orthoptists). For the application example in a German setting, data from the published medical literature were used. RESULTS: In the base-case analysis of the application example, screening high-risk children by ophthalmologists had the lowest average cost per case detected but became dominated (less effective and more costly than an alternative) if a low (5.3%) probability of familial clustering of strabismus was assumed. Considering the various assumptions tested in the sensitivity analysis, screening of all children up to the age of 1 year by ophthalmologists was the only strategy not dominated by others. Detection rates, including cases detected before screening, were between 72% and 78% for the strategies that screen for all children. CONCLUSIONS: The model suggests that in Germany, both from a cost-effectiveness and a pure effectiveness point of view, screening all children up to the age of 1 year by ophthalmologists is the preferred strategy to detect amblyopia or amblyogenic factors. All strategies left a significant portion of children undetected. PMID- 12637839 TI - Impact of public support payments, intensive psychiatric community care, and program fidelity on employment outcomes for people with severe mental illness. AB - This study explored the relationship of public support payments, intensive psychiatric community care (IPCC), and fidelity of implementation to 1-year employment outcomes for 520 veterans with severe mental illness (SMI) in a clinical trial of IPCC. At study entry, 455 (87.5%) participants received public support. At 1 year, 46 (8.8%) participants met criteria to be classified as workers. A multivariate analysis indicated that baseline public support was significantly associated with a lower likelihood of employment, and baseline work was positively associated with employment at 1 year. IPCC patients were three times more likely to be working than control subjects, and a significant interaction favored well-implemented IPCC programs over others. This study points out not only the inhibiting effect of public support payment on employment but also the value of IPCC and the special importance of fidelity to program models for employment for people with SMI. PMID- 12637840 TI - Utility of the Time-Line Follow-Back to assess substance use among homeless adults. AB - Assessing substance use of homeless persons is a critical task. This study examines the test-retest reliability, concurrent validity, and sensitivity to change of the Time-Line Follow-Back interview, a calendar instrument used to assess days and quantities of alcohol use and days of illicit drug use, in the multisite Collaborative Program to Prevent Homelessness (CPPH). The Time-Line Follow-Back was reliable for assessing use during the past month and the recent 6 months. Results from the Time-Line Follow-Back were correlated with other self reports of use, with research diagnoses of substance use disorder, and with clinician ratings of severity of substance abuse. The Time-Line Follow-Back detected changes in clients with severe mental illness and in those with less severe psychiatric problems. PMID- 12637841 TI - Long-term psychological outcome for non-treatment-seeking earthquake survivors in Turkey. AB - This study examined the incidence of posttraumatic stress disorder (PTSD) and depression in 586 earthquake survivors living in prefabricated housing sites a mean of 20 months after the 1999 earthquake in Turkey. The estimated rates of PTSD and major depression were 39% and 18%, respectively. More severe PTSD symptoms related to greater fear during the earthquake, female gender, older age, participation in rescue work, having been trapped under rubble, and personal history of psychiatric illness. More severe depression symptoms related to older age, loss of close ones, single marital status, past psychiatric illness, previous trauma experience, female gender, and family history of psychiatric illness. These findings suggest that catastrophic earthquakes have long-term psychological consequences, particularly for survivors with high levels of trauma exposure. These findings lend further support to the need for long-term mental health care policies for earthquake survivors. Outreach service delivery programs are needed to access non-treatment-seeking survivors with chronic PTSD. PMID- 12637842 TI - Treatment preferences and determinants in victims of sexual and physical assault. AB - To determine the treatment preferences (, medication, counseling, or both) for victims of recent trauma at risk for posttraumatic stress disorder and the determinants of those choices, preferences were elicited from 466 consecutive victims of physical or sexual assault at an urban emergency department as part of the routine clinical evaluation by emergency department social workers. Demographics, assault characteristics, and clinical history were used to predict preference. More than 80% of victims expressed an interest in treatment, with more interested in counseling (76%) than medication (62%). Female gender and assaults of a sexual nature were most highly predictive of preference for medication, whereas female gender, sexual assault, a history of psychiatric treatment, and perceived life threat from the trauma were most predictive of preference for counseling and preference for any treatment. Because treatment preferences were easily obtained from assault victims seeking emergency care, the elicitation of these treatment preferences and the consideration of them in the treatment planning process could facilitate engagement in treatment and overcome the avoidance of treatment by some victims. PMID- 12637844 TI - Differential impairments of facial affect priming in subjects with acute or partially remitted major depressive episodes. AB - Eleven subjects with acute major depressive episodes and 9 subjects with partially remitted major depressive episodes were compared with 24 healthy control subjects on an emotional priming task. Positive and negative emotional facial expressions were presented as subthreshold primes, followed by a neutral pattern mask, and an emotionally neutral face as suprathreshold target. Subjects had to judge if they had seen a pleasant or an unpleasant facial expression. Healthy subjects and subjects with partially remitted depression judged the neutral target as significantly more unpleasant when negative emotional facial expressions were presented as primes as compared with when positive facial expressions were presented as primes. In contrast, subjects with acute depression did not show a significant judgment shift. It is concluded that subjects with acute depression are not able to preactivate emotional concepts by subthreshold presented emotional facial expressions. PMID- 12637843 TI - American attitudes toward and willingness to use psychiatric medications. AB - Despite recent advances in treatment, many Americans decline to take prescribed psychiatric medication. This study explores the role of attitudes regarding the effectiveness of and potential problems associated with psychiatric medications on Americans' willingness to use them. Face-to-face interviews of a US household population sample were done with 1387 volunteers. The 1998 General Social Survey's (response rate, 76.4%) included questions about efficacy, problems, and potential use. Most Americans agree that psychiatric medications are effective, and fewer than half had concerns regarding potential problems. However, the majority of respondents would not be willing to take them. Willingness to use is influenced by these attitudes and other factors, including health status and past use of mental health treatments. Although Americans perceive psychiatric medications to be effective, and this influences their willingness to take them, many still are not willing to take them. PMID- 12637846 TI - The influence of litigation on maintenance of posttraumatic stress disorder. PMID- 12637845 TI - Dissociative disorders and traumatic childhood experiences in transsexuals. AB - In this first prevalence study of dissociative symptoms and different forms of childhood experiences among transsexuals, 41 transsexuals and 115 psychiatric inpatients were compared by means of the Interview for Dissociative Disorders (SCID-D-R), the Dissociative Experiences Scale (DES), and the Childhood Trauma Questionnaire (CTQ). The total score for the dissociative symptoms revealed no significant differences between the transsexuals and the psychiatric inpatients. However, the higher DES score among transsexuals compared with a normal population was found to be due largely to one item. A surprisingly high prevalence of emotional maltreatment was recorded. The results suggest that both the DES and the SCID-D-R have limited validity as instruments for screening and diagnosing dissociative disorders in transsexuals. Psychiatrists should be mindful of the possible existence of dissociative disorders in transsexual patients. Further investigations are needed to clarify the effects of traumatic childhood experiences on sexual identity in transsexuals and to throw more light on the phenomenological correlation between transsexualism and dissociative identity, using taxometric analyses. PMID- 12637847 TI - Suicidal ideation and quality of life among adult Brazilian outpatients with depressive disorders. PMID- 12637848 TI - Anorexia nervosa subtypes: differences in recovery. PMID- 12637849 TI - Obsessive-compulsive disorder and Jewish religiosity. PMID- 12637851 TI - The stigma of emergency medicine. PMID- 12637850 TI - Phenomenology of visual hallucinations in psychiatric conditions. PMID- 12637852 TI - Time savings with prehospital thrombolysis in an urban area. AB - The aim of this study was to evaluate the time saved by prehospital thrombolysis compared with the usual inhospital reperfusion strategies in Geneva, Switzerland, a moderate-sized urban area with short transportation times (median time 4 min). Patients with acute myocardial infarction, diagnosed clinically and by means of a 12-lead electrocardiogram in a mobile intensive care unit, without contraindications to thrombolysis, were randomly assigned either to 'prehospital thrombolysis' or to 'usual hospital care', i.e. inhospital reperfusion strategies (thrombolysis or primary angioplasty). Sixteen patients with similar clinical characteristics were randomly selected. The mean time between the arrival of the physician by the patient and the initiation of therapy was shorter for 'prehospital thrombolysis' (26.0+/-10.3 min) than for 'usual hospital care' (94.93+/-6.3 min, P=0.0004). Time savings were even greater when time between the beginning of symptoms and the initiation of therapy was analysed (152.6+/-76.6 versus 251.2+/-86.3 min, P=0.0004). We concluded that, even in an urban area with a readily available catheterization laboratory, prehospital thrombolysis can be initiated safely immediately after the diagnosis of acute myocardial infarction, and is time saving. PMID- 12637853 TI - Quality assurance with regard to outcome and use of medical resources for patients hospitalized with acute chest pain: a comparison between a city university hospital and a county hospital. AB - This study aimed to test the hypothesis that there is a difference in mortality between patients hospitalized with acute chest pain in a university hospital and those hospitalized in a county hospital, and to describe differences in characteristics and use of medical resources in these two settings. All patients hospitalized at Sahlgrenska University Hospital in Goteborg (with a catchment population of 706 inhabitants/km(2)) and Uddevalla County Hospital (with a catchment population of 34 inhabitants/km(2)) with symptoms of acute chest pain during a registration period of 6 months were included in the study. A total of 1592 patients in the city hospital and 822 in the county hospital fulfilled the given criteria for inclusion. Patients in the urban area differed from those in the rural area in that they had a lower prevalence of previous angina pectoris and hypertension and a higher prevalence of previous cancer, previous percutaneous transluminal coronary angioplasty (PTCA) and coronary artery bypass grafting (CABG) and current smoking. On admission to hospital, patients in the urban area less frequently showed clinical signs of congestive heart failure and acute ischaemia on the electrocardiogram (ECG) but more frequently had a pathological ECG without signs of ischaemia and more frequently had a heart rate >100 beats/min. The use of medical resources differed between the two hospitals. Revascularization was more frequent in the city hospital and the use of -blockers in the county hospital. The overall 30 day mortality was 4.7% in the urban area and 4.3% in the rural area (P=0.74). When correcting for differences at baseline, the risk ratio for death in the county hospital versus the city hospital was 0.84 (95% confidence interval 0.51-1.40, P=0.53). In conclusion, among patients hospitalized with acute chest pain in a city university and a county hospital the mortality during the subsequent 30 days did not differ. However, there were differences in terms of the use of medical resources and in previous history, chronic medication prior to hospital admission and status on admission between the two cohorts. PMID- 12637854 TI - Is a career in emergency medicine associated with stigma? AB - OBJECTIVE: Many emergency medicine staff report anecdotally that fellow hospital staff have a low opinion of emergency medicine. No research into this attitude has been published. The aim of this study is to determine whether there is stigma attached to emergency medicine and practitioners. METHOD: A postal questionnaire of all medical staff at a district general hospital, to evaluate the presence or absence of eight perceptions associated with stigma. RESULTS: The response rate was 49.5%, with the response rate decreasing with decreasing grade. Of the stigmatizing themes tested in this study, six of the eight were demonstrated to be associated with negative attitudes, with the remaining two themes positive attitudes towards emergency medicine were suggested. The responses were similar, irrespective of grade and speciality (where given by respondees). Responses that were ambivalent or not completed varied between 25 and 48% in the returned questionnaires. CONCLUSIONS: This paper demonstrates that stigmatizing opinions towards emergency medicine exist and that these negative opinions may be widely held by hospital staff. PMID- 12637855 TI - Airway equipment in Scottish emergency departments. AB - Airway management is the cornerstone of resuscitation. Emergency endotracheal intubation is more likely to be difficult in the accident and emergency (A&E) department setting compared with the operating room. A&E departments must have an appropriate selection of equipment to deal with difficult airway problems. The aim of this study was to determine the type and range of equipment for airway management in A&E departments in Scotland, UK. A two-page postal survey was sent to consultants in charge of 24 A&E departments covering 98% of major trauma patients in Scotland. The response rate was 96% (23/24). Every department had equipment for basic airway management and all had laryngoscopes, stylets, gum elastic bougies and capnography. Ninety-six per cent (22/23) had equipment for performing a surgical airway and 74% (17/23) possessed laryngeal mask airways suitable for adults; only one department did not possess a suitable rescue device. Thirty per cent (7/23) of departments did not use capnography routinely to confirm correct placement of endotracheal tubes. It is concluded that airway equipment in Scottish A&E departments is adequate for basic airway care and endotracheal intubation. Nearly all departments have access to a suitable rescue device for the failed or difficult airway. Capnographic confirmation of tube placement should be mandatory in A&E. PMID- 12637857 TI - Mushroom poisoning: an analysis of the data between 1996 and 2000. AB - One hundred and forty-three cases (89 women and 54 men) of mushroom poisoning recorded at the emergency service of Osmangazi University Hospital, Faculty of Medicine, between 1996 and 2000 were evaluated. The first symptoms seen were loss of consciousness, fatigue, dizziness, severe headaches, abdominal discomfort and vomiting. The symptoms characteristically appeared suddenly. Eight patients suffering from poisoning caused by cultivated mushrooms, and four patients suffering from poisoning caused by wild mushrooms died from fulminant hepatic failure. The other patients were discharged within a period of 1-10 days. It is suggested that people should be informed of the possibility of mushroom poisoning, which has been increasing recently in Turkey. PMID- 12637856 TI - Importance of early identification of methylenedioxymethamphetamine ('ecstasy') ingestion in victims of motor vehicle accidents. AB - The blood of motor vehicle accident victims is routinely screened upon their arrival at the emergency services mainly for alcohol, unless the suspicion of a specific compound arises. Two young men who sustained severe internal and orthopaedic injuries after a motor vehicle accident are described. The conscious patient denied their having used stupefacients, but toxicological analysis upon arrival at the operating room detected methylenedioxymethamphetamine, the metabolite methylenedioxyamphetamine and methamphetamine in their blood and urine specimens. Methylenedioxymethamphetamine concentrations in the clotted blood and in the urine ranged between 0.9-1.15 and 55-70 mg/l, respectively. Methylenedioxyamphetamine concentrations for both patients were less than 0.2 mg/l in the blood and 2.0-3.0 mg/l in the urine. Each had a blood methamphetamine concentration greater than 250 ng/ml. There was no trace of alcohol. Three days after their arrival at the hospital, acute liver insufficiency and mild rhabdomyolysis (serum glutamate-pyruvate transaminase/serum glutamic-oxaloacetic transaminase 1245/218 mU/ml, creatine phosphokinase 48 000 U/ml, respectively) were diagnosed in both patients. Appropriate treatment was administered in an intensive care area and both were discharged home several weeks later without sequelae. These findings suggest that in this era of the widespread abuse of 'ecstasy', concentrations of methylenedioxymethamphetamine, methylenedioxyamphetamine or metamphetamine should be sought routinely in motor vehicle accident victims admitted to emergency services with an altered state of consciousness so that the early monitoring of the potential development of organ pathology can be implemented. PMID- 12637858 TI - Paragliding accidents--the spine is at risk. A study from a Swiss Trauma Centre. AB - In recent decades, paragliding-like other fashionable activities-has become a part of lifestyle and outdoor activities. The introduction of protective devices has helped to reduce the risk of severe injuries. However, it seems that the spine remains the paraglider's 'Achilles heel'. Better education, training, and the introduction of innovative back protectors are required to reduce the frequency and severity of paragliding injuries. PMID- 12637859 TI - Maxillofacial injuries in North American vehicle crashes. AB - Twenty cases of maxillofacial trauma to vehicle occupants in motor crashes were retrospectively analysed using data from the CIREN project (Crash Injury Research Engineering Network - a collaboration between eight level 1 trauma centres in America). The facial fractures and lacerations were classified and a correlation was made between the deformation of the vehicle and the injuries sustained. The importance of airbags as supplemental restraint systems is emphasized, as is the necessity for coexistent seatbelt use. The vehicle 'A' pillar as a source of injury is highlighted. Although there is no doubt that airbags save lives, the possible risks of powerful airbag deployment to small stature, out of position, vehicle occupants is discussed. PMID- 12637860 TI - Pulmonary oedema in SCUBA divers: pathophysiology and computed risk analysis. AB - SCUBA diving has become a popular sport, with an increasing number of people participating in it. Although it is an essentially very safe activity, several specific medical problems are associated with diving. The present paper addresses diving-related pulmonary oedema, which is a rarely reported condition. It has been described mostly in cold water diving, and its occurrence in warm water has not been documented before. We delineate the pathophysiology of this condition and its treatment, and use a novel computational model to analyse further the mechanisms leading to diving-related pulmonary oedema. A better understanding of diving-related pulmonary oedema may lead to earlier recognition and treatment, and, potentially, to its prevention. The physiological mechanisms likely to lead to diving-related pulmonary oedema are well recognized. Consequently, we could design and construct a bio-mechanical computer model of the alveolar septa to explore the pathophysiology of diving-related pulmonary oedema and the vulnerability of individual divers as they relate to some mechanical characteristics of their lung structure. The physiological mechanisms of diving related pulmonary oedema and the results provided by the computational model successfully delineated the process. The model predicted that the risk of injury is significantly increased in individuals who have a stiffer lung parenchyma or lower lung compliance values. PMID- 12637861 TI - Ruptured tibialis posterior tendon, presenting as a lateral ligament sprain. PMID- 12637862 TI - Acute massive rhabdomyolysis due to prolonged inhalation of liquid gas. AB - A very rare case of non-fatal acute massive rhabdomyolysis caused by unintentional prolonged inhalation of liquid gas (consisting of butane and propane) in a previously healthy adult is presented. The immediate diagnosis and intensive symptomatic therapy prevented any other severe complications of rhabdomyolysis, and the patient made a complete recovery without any sequelae. PMID- 12637863 TI - Spontaneous massive haemopneumothorax: case reports. AB - Spontaneous haemopneumothorax is an infrequently reported entity. We report here on two patients with spontaneous haemopneumothorax complicated by massive haemorrhage. Sources of bleeding were caused by torn pleural adhesion and ruptured apical bullae. Inappropriate management can lead to an increase in mortality and morbidity. We emphasize the importance of close monitoring, early recognition, prompt resuscitation and thoracotomy because of the possible lethal implications. PMID- 12637864 TI - The brain-dead patient or a flower in the vase? The emergency department approach to the preservation of the organ donor. AB - Transplantation has been incorporated into the treatment of patients with end stage diseases of most major organ systems in recent years. However, organ supply is the greatest limitation to organ transplantation. Among the factors that can enhance organ supply, donor management has received the least attention. The importance of establishing an acceptable method of rapidly and accurately determining brain death in potential donors cannot be overemphasized. With an increased awareness of donor management issues and the application of a rational physiological approach, the supply of functional organs for transplantation can be increased. Rapid and continuing resuscitation of clinically brain-dead trauma victims is mandatory. This review addresses the evaluation and management of the organ donor within the emergency department. Common management problems (hypotension, arrhythmias, diabetes insipidus, oliguria, and coagulopathy) are discussed in detail. An aggressive, proactive approach to the medical management of the potential donor is recommended in order to limit the number of medical failures and maximize the number of organs donated. PMID- 12637865 TI - Questionnaire reveals variations in the management of acute first time shoulder dislocations in the Netherlands. AB - AIM: To collect information about the management of patients with acute first time shoulder dislocation by (orthopaedic) surgeons in the accident and emergency departments of Dutch hospitals. DESIGN: Questionnaires were sent to 131 (orthopaedic) surgeons of 74 Dutch hospitals. METHOD: We enquired whether patients with acute first time shoulder dislocation are managed according to protocols in accident and emergency departments; which radiographs are deemed necessary before and after reduction; which reduction methods are generally performed; and what kind of pain relief or sedatives are generally administered. Furthermore, we enquired into the method and duration of immobilization after reduction. RESULTS: The response rate was 73%. Sixty-five per cent of the hospitals are used to managing acute first time shoulder dislocation according to protocols. Making prereduction and postreduction radiographs is standard practice in almost all hospitals. The most favoured methods of reduction are those according to Kocher and Hippocrates. In 85% of cases sedatives are administered before reduction, and in addition to that systemic analgesia is administered in 30%, whereas intra-articular analgesia is preferred in 25% of cases. CONCLUSION: Apparently, a protocol for the management of acute first time shoulder dislocation is not standardly available in all hospitals. On the basis of current literature and the results of this questionnaire it may be useful to establish a guideline for diagnostic procedures and the management of acute first time shoulder dislocation in accident and emergency departments. PMID- 12637866 TI - Zyban: increasing the workload in an accident and emergency department? PMID- 12637867 TI - Risk score for interhospitalary transport. PMID- 12637868 TI - Application of guidelines for severe head trauma: data from an Italian database. PMID- 12637869 TI - Pulmonary embolism challenges. PMID- 12637870 TI - [Therapeutic management of hepatic metastases from colorectal cancers]. PMID- 12637871 TI - [What exams should be ordered for the pretherapeutic work-up? Criteria of quality and expected results]. PMID- 12637872 TI - [When is first-line resection of hepatic metastasis indicated?]. PMID- 12637873 TI - [What are the roles of neoadjuvant, adjuvant and palliative chemotherapy in the management of hepatic metastasis of colorectal origin?]. PMID- 12637874 TI - [Modalities and feasibility of treatment by local destruction of hepatic matastases (TLDHM)]. PMID- 12637875 TI - [Can initially non-respectable hepatic metastases be made resectable?]. PMID- 12637883 TI - [MR Sialography]. AB - Indications for sialography have became rare and radiologists are less and less experienced for salivary gland cannulation. MR Sialography allows opacification of salivary ducts without any cannulation and could replace residual indications for sialography. The purpose of this paper is to review the principle of the technique, the normal and pathologic aspects of MR Sialography and to emphasize advantages and limitations compared to other techniques. PMID- 12637882 TI - [Lower limb arterial disease: Doppler sonography follow-up after revascularization]. AB - Follow-up color duplex sonography after arterial surgery or angioplasty for lower limb arterial disease evaluates the outcome of the procedure and searches for lesions compromising patency. The various types of lesions are described. After surgical bypass, lesions include stenosis, anastomotic abnormality (enlargement, false-aneurysm), arterio-venous fistula, intrinsic bypass abnormalities, and collections. After endoluminal treatment, the main lesions include residual stenosis, restenosis, and in-stent hyperplasia. Local complications related to arterial puncture are described. The frequency and the type of lesions encountered depend on the interval between the date of the revascularization and the follow-up examination. PMID- 12637884 TI - [Small bowel obstruction from adhesions: which CT severity criteria to research?]. AB - PURPOSE: To determine the value of known computed tomographic (CT) criteria to differentiate non-complicated from complicated (strangulation, necrosis) small bowel obstruction. MATERIALS AND METHODS: 43 patients with a definitive diagnosis of small bowel obstruction based on clinical, sonographic, CT, surgical and pathological findings were included. All patients had small bowel obstruction caused by adhesions confirmed at surgery. The obstruction was non-complicated in 28 patients and complicated in 15 patients. The CT examinations from all patients were retrospectively reviewed by three experienced radiologists using a set of pre-defined criteria. Attention was focused on the following signs: reduced enhancement of the small bowel wall, mural thickening, congestion of small mesenteric veins, and ascites. Results were correlated with surgical and/or pathological data. RESULTS: For the diagnosis of complicated obstruction, reduced bowel wall enhancement had a sensitivity of 57% and a specificity of 100%, a bowel wall thickness greater than 3 mm had a sensitivity of 35% and a specificity of 100% and a bowel wall thickness less than 1 mm had a sensitivity of 35% and a specificity of 93%. Ascites and congestion of small mesenteric veins were not significant. The multivariate analysis showed that the association of bowel-wall thickening and reduced enhancement of the small bowel wall was significant (sensitivity of 71%, specificity 100%, and accuracy 90%). CONCLUSION: Among the CT criteria used to diagnose complications from small-bowel obstruction that were evaluated in this study, only three were significant with a high specificity but low sensitivity. PMID- 12637886 TI - [Cost efficiency study of lacrimal canal obstruction treatment]. AB - OBJECTIVE: To compare three methods used to clear the lacrimal canal. PATIENTS AND METHODS: A total of 32 files (20 for the dacryocysto-rhinostomy by endoscopic way, 5 for the dacryocysto-rhinostomy by external way and 7 cases of percutaneous stenting) were reviewed. For each, costs of performed interventions were reviewed. An average of the total cost per patient was calculated for all three methods and was then correlated to the initial efficiency of the method. RESULTS: Results show that the average cost for stenting is up to 1,366.80 EUR, for the endoscopic way is up to 2 001.53 EUR and for the external way is up to 2,220.46 EUR. ANOVA (Analysis of Variance) gives significant results when comparing stent to endoscopic way (p=0.000007) and to the external way (p =0.02). The analysis of sensitivity concerning the success rate of various treatments shows that the stent was potentially the most cost-effective and that a failure rate of 35.1% is rather acceptable by comparison to the endoscopic method. If the stent is considered as an outpatient intervention, then, a failure rate of 50% is acceptable. CONCLUSION: If the failure rate stays under 35.1%, the stent may be considered as a short-term cost-effective method. PMID- 12637885 TI - [Value of MRI in symptomatic patients treated with tamoxifen]. AB - PURPOSE: To assess the MR imaging (MRI) findings in symptomatic tamoxifen treated women with abnormal transvaginal sonography. PATIENTS AND METHODS: From january 1997 to june 2000, 32 consecutive symptomatic tamoxifen treated-women with abnormal transvaginal sonography were prospectively studied by MRI. T1-weighted, T2-weighted, post-contrast T1-weighted and dynamic gradient-echo T1-weighted sequences were used. All patients underwent uterine sampling within one month of MRI. RESULTS: Endometrial thickness at sonography ranged from 5 to 48 mm (mean thickness 19 mm), and on T2-weighted imaging ranged from 3 to 50 mm (mean=25 mm). Three MRI patterns were found. Pattern 1 (13 patients) was defined as homogeneous high signal intensity of the endometrium on T2W images, and signal void in the lumen on gadolinium-enhanced images. Pattern 2 (8 patients) was defined as heterogeneous endometrial signal on T2W images, and latticelike enhancement traversing the endometrial canal on gadolinium-enhanced images. Pattern 3 (11 patients) was defined as heterogeneous signal on T2W images with masses or nodules which were better seen on dynamic gadolinium-enhanced images. In pattern 1 we found 13 atrophic endometrium, in addition there were 4 polypoid glandulo cystic proliferation (PGCP), and 1 adenomyosis. In pattern 2 we found 3 PGCP, 4 atrophy and 1 polyp without hyperplasia. The 2 carcinomas and the polyps with hyperplasia were found in pattern 3 (11 patients). CONCLUSION: In our experience MRI allows differentiation of lesions which may require surgery from other lesions in which noninvasive follow-up is possible. PMID- 12637887 TI - [Small bowel diverticulitis: the role of CT]. AB - The authors report a case of small bowel diverticulitis diagnosed by computed tomography. They describe the CT findings and review its advantages for diagnosis of this uncommon entity that is rarely suspected at physical examination. PMID- 12637888 TI - [Acute spinal epidural and subdural hematomas]. AB - Acute spinal epidural and subdural hematomas are rare entities. The clinical presentation is similar in both cases. MR imaging is the most appropriate imaging technique. On axial images, epidural hematoma has a biconvex shape. It is separated from the spinal cord by the hypointense dura on T2-weighted images. Conversely, subdural hematoma has a crescentic shape on axial images and is located medially to the dura. In the latter case, the epidural fat is not involved. There is no surgical indication for patients with early and rapid regressive symptoms. In other cases the surgical treatment consists of hematoma evacuation. PMID- 12637889 TI - [Radiological spectrum of myositis ossificans circumscripta evolution]. AB - Myositis ossificans circumscripta is a benign pathology of soft tissue occurring in young patients often after localized trauma. Histological and radiological appearances may mimic a malignant neoplasm, mainly sarcoma. We report a case characteristic of myositis ossificans circumscripta, to illustrate diagnostic arguments which are based on their appearance on conventional radiographs, computed tomography and magnetic resonance imaging and on their changes with time. PMID- 12637890 TI - [Paraganglioma and pregnancy]. AB - A paraganglioma is an extra-adrenal pheochromocytoma, rarely observed during pregnancy. Maternal prognosis depends on early diagnosis and multidisciplinary management prior to tumor resection. The tumor is localized by ultrasonography or MRI. During the post-partum, and if MRI is not available, CT is often sufficient. We report a paragangliona in a patient with intrauterine fetal demise at 27 weeks gestation diagnosed by ultrasonography and computed tomography, who underwent surgical resection. PMID- 12637892 TI - Montelukast (singulair) for allergic rhinitis. PMID- 12637891 TI - [Idiopathic epiphoras in adults treated with lacrymal-nasal stents]. AB - Fluoroscopic nonsurgical placement of an expandable polyurethane stent is less invasive than surgery for the treatment of epiphora due to obstruction of the nasolacrimal duct. It needs to be performed with the collaboration of an ophthalmologist, using a high resolution digital subtraction imaging unit. PMID- 12637893 TI - Anastrozole (Arimidex) vs. tamoxifen for treatment of early breast cancer. PMID- 12637894 TI - Drug-eluting stents. PMID- 12637895 TI - Camouflage might not mean compromise. PMID- 12637896 TI - Genetic predisposition to external apical root resorption. AB - External apical root resorption (EARR) can be an undesirable sequela of orthodontic treatment. Previous studies have suggested that EARR has a substantial genetic component. Linkage and association were examined between polymorphisms of the interleukin IL-1 (IL-1A and IL-1B) genes and EARR in 35 white American families. Buccal swab cells were collected for DNA isolation and analysis. The EARR in the maxillary central incisors, the mandibular central incisors, and the mesial and distal roots of the mandibular first molar were analyzed separately and together by using both linkage and association methods of analysis. Highly significant (P =.0003) evidence of linkage disequilibrium of IL 1B polymorphism with the clinical manifestation of EARR was obtained. The analysis indicates that the IL-1B polymorphism accounts for 15% of the total variation of maxillary incisor EARR. Persons homozygous for the IL-1B allele 1 have a 5.6 fold (95% CI 1.9-21.2) increased risk of EARR greater than 2 mm as compared with those who are not homozygous for the IL-1 beta allele 1. Data indicate that allele 1 at the IL-1B gene, known to decrease the production of IL 1 cytokine in vivo, significantly increases the risk of EARR. These findings are consistent with an interpretation of EARR as a complex condition influenced by many factors, with the IL-1B gene contributing an important predisposition to this common problem. Defining genetic contributions to EARR is an important factor in understanding the contribution of environmental factors, such as habits and therapeutic biomechanics. PMID- 12637897 TI - Peg-shaped and small lateral incisors not at higher risk for root resorption. AB - The purpose of this study was to examine the pattern of external root resorption for peg-shaped and small lateral incisors according to various conditions and to compare it with that of normal-shaped lateral incisors. The periapical radiographs of 114 patients with 60 peg-shaped and 54 small lateral incisors were measured before and after treatment. Crown shape was assessed by examining pretreatment study casts. Findings showed that there was no significant difference in external root resorption between peg-shaped (1.09 mm) and normal lateral incisors (0.88 mm). However, a statistically significant difference in apical root resorption between small lateral incisors (1.03 mm) and normal lateral incisors (1.62 mm) was found. Peg-shaped laterals were also coincidentally found to occur more frequently on the left side of the arch. PMID- 12637898 TI - Ectopic eruption and severe root resorption. AB - The treatment of a patient with maxillary canine-central incisor transposition is presented. Treatment options were limited because the roots of the maxillary right lateral and central incisors had severe resorption and generalized shortening. The multidisciplinary approach and treatment sequence used to achieve an esthetic and functional result are discussed in this case report. PMID- 12637899 TI - Long-term follow-up of Class II adults treated with orthodontic camouflage: a comparison with orthognathic surgery outcomes. AB - Thirty-one adults who had been treated with orthodontics alone for Class II malocclusions were recalled at least 5 years posttreatment to evaluate cephalometric and occlusal stability and also their satisfaction with treatment outcomes. The data were compared with similar data for long-term outcomes in patients with more severe Class II problems who had surgical correction with mandibular advancement, maxillary impaction, or a combination of those. In the camouflage patients, small mean changes in skeletal landmark positions occurred in the long term, but the changes were generally much smaller than in the surgery patients. The percentages of patients with a long-term increase in overbite were almost identical in the orthodontic and surgery groups, but the surgery patients were nearly twice as likely to have a long-term increase in overjet. The patients' perceptions of outcomes were highly positive in both the orthodontic and the surgical groups. The orthodontics-only (camouflage) patients reported fewer functional or temporomandibular joint problems than did the surgery patients and had similar reports of overall satisfaction with treatment, but patients who had their mandibles advanced were significantly more positive about their dentofacial images. PMID- 12637900 TI - Predictive value of morphologic parameters for successful correction of Class II Division 2 malocclusion. AB - The aim of this study was to assess the predictive value of pretreatment cephalometric parameters and apical base size for successful correction of Angle Class II Division 2 malocclusion. Pretreatment lateral cephalograms and pretreatment and posttreatment study casts of 96 subjects with Class II Division 2 malocclusions were examined to obtain 23 cephalometric parameters and to measure the size of the apical bases. Success of occlusal correction was evaluated as the percentage of change of peer assessment rating scores during treatment; percentage of change was the dependent variable in multivariate statistical analyses used to test the predictive value of the assessed parameters. Cephalometric parameters of pretreatment craniofacial morphology were insignificant as predictors of successful orthodontic correction of the malocclusions. Maxillary apical base size was the strongest predictor of occlusal correction and might serve as a valuable diagnostic predictor of successful treatment outcome. PMID- 12637901 TI - Treatment effects of the mandibular anterior repositioning appliance on patients with Class II malocclusion. AB - The mandibular anterior repositioning appliance (MARA) is a tooth-borne functional appliance for use in patients with Class II malocclusions; it positions the mandible forward into a Class I occlusion. The aim of this study was to investigate the MARA's dental and skeletal effects on anterior, posterior, and vertical changes in 30 Class II patients. The treatment group consisted of 12 boys with an average age of 11.2 years and 18 girls with an average age of 11.3 years. A pretreatment cephalometric radiograph was taken 2 weeks before treatment, and a posttreatment cephalometric radiograph was taken 6 weeks after removal of the MARA, with an average treatment time of 10.7 months. The mean and standard deviation were calculated for each cephalometric variable, and Student t tests were performed to determine the statistical significance of the changes. The results of the study showed that the MARA produced measurable treatment effects on the skeletal and dental elements of the craniofacial complex. These effects included a considerable distalization of the maxillary molar, a measurable forward movement of the mandibular molar and incisor, a significant increase in mandibular length, and an increase in posterior face height. The effects of the MARA treatment were then compared with those of the Herbst and Frankel appliances. The treatment results of the MARA were very similar to those produced by the Herbst appliance but with less headgear effect on the maxilla and less mandibular incisor proclination than observed in the Herbst treatment group. PMID- 12637902 TI - Activator versus cervical headgear: superimpositional cephalometric comparison. AB - Clinical trials comparing activator and headgear treatment have shown comparable effectiveness in the total result achieved, but the mechanism of correction is still uncertain. Most studies have used conventional cephalometric methods to evaluate treatment effects, and this might be a factor for the inconclusive results. The aim of this retrospective investigation was to compare the effects of activator and cervical headgear treatment with a superimpositional cephalometric method that could discern between vertical and horizontal effects as well as skeletal, dental, and rotational treatment results. The sample consisted of 2 groups of Class II Division 1 patients, treated without extraction by the same clinician (22 patients were treated with a modified activator-type functional appliance, and 30 patients were treated with a combination of cervical headgear and fixed edgewise appliances). Lateral cephalometric radiographs taken at the beginning of treatment and after Class II molar correction were evaluated conventionally and with a superimpositional method. Regarding the conventional cephalometric measurements, the only difference in the anteroposterior dimension between the 2 treatment modalities was the significantly reduced SNA angle in the headgear group. Both appliances appeared to produce minimal changes in FMA and GoGn-SN angles, and there were no statistically significant differences between the treatment groups. Regional superimpositions showed differences in the movement of molars: the maxillary molar was found to move more posteriorly and inferiorly in the headgear group. Conversely, the mandibular molar was found to move toward the occlusal plane more in the activator group. Assessment of mandibular skeletal changes showed that the mandible moved anteriorly by approximately 1 mm more in the activator than in the headgear group. The overall effect of the 2 appliances was found to be clinically comparable. However, the individual components of change showed differences characteristic of each appliance. PMID- 12637903 TI - Long-term effects of Class III treatment with rapid maxillary expansion and facemask therapy followed by fixed appliances. AB - In this cephalometric investigation, we compared the long-term effects of an initial phase of rapid maxillary expansion and facemask (RME/FM) therapy followed by comprehensive edgewise therapy with the effects of growth in untreated, matched controls. The treated sample consisted of 34 patients who underwent RME/FM treatment before the pubertal growth spurt (average age, 8 years 3 months at the beginning of treatment). At the final observation period (average age, 14 years 10 months), all patients were in decelerative growth phases as determined by the cervical vertebral maturation (CVM) method. After the first 10 months of active treatment, significant favorable changes in both the maxillary and the mandibular skeletal components were noted. The forward movement of the maxilla was 1.8 mm greater than in the controls, mandibular projection was reduced by almost 3 mm, and the relative sagittal intermaxillary discrepancy improved by 4.3 mm, as measured by the Wits appraisal. During the posttreatment period, the treated and untreated Class III subjects generally grew similarly, although the skeletal relationship of the maxilla to the mandible remained unchanged in the RME/FM group, whereas the controls had an increased skeletal discrepancy of 3.0 mm. Over the long term, there was a slightly greater increase in midfacial length (1.6 mm) in the treatment group than in the controls. Similarly, the distance from Point A to nasion perpendicular decreased by 1.2 mm in the treated group. The overall increase in mandibular length was 2.4 mm less in the RME/FM group than in the controls, and mandibular projection relative to nasion perpendicular was 3.0 mm less in the treated group. The change in the Wits appraisal was substantial between groups (6.1 mm), with an improvement in the intermaxillary relationship in the treated group (3.4 mm); the Wits appraisal worsened (-2.7 mm) in the untreated controls. No clinically significant differences were observed between the groups in the vertical dimension. Overjet increased significantly in the treated group relative to the controls (4.4 mm), whereas the molar relationship decreased significantly (-3.9 mm). It appears that the favorable skeletal change observed over the long term is due almost entirely to the orthopedic correction achieved during the RME/FM protocol. During the posttreatment period that includes the pubertal growth spurt, craniofacial growth in RME/FM patients is similar to that of untreated Class III controls. Aggressive over-correction of the Class III skeletal malocclusion, even toward a Class II occlusal relationship, appears to be advisable, with the establishment of positive overbite and overjet relationships essential to the long-term stability of the treatment outcome. PMID- 12637904 TI - Comparison of skeletal and dental changes between 2-point and 4-point rapid palatal expanders. AB - Rapid palatal expansion has been a clinically accepted technique used by orthodontists for over 100 years. Its primary goal is to maximize orthopedic and minimize orthodontic movements of teeth. Historically, this was best accomplished by including 4 teeth in the appliance. However, including more teeth makes construction and insertion more difficult. The appliance also is less comfortable for patients and hinders oral hygiene. The 4-point hyrax expander can be modified by removing the 2 anterior wires, creating a 2-point expander between the first molars only. The purpose of this study was to determine the difference, if any, between midpalatal suture separation and dental expansion produced between 2 point and 4-point palatal expanders. Thirty subjects between the ages of 6 and 16 years were randomly assigned to either the 2-point (n = 15) or the 4-point group (n = 15). The groups were compared on dental and radiographic landmarks. The groups showed no statistical differences in total molar cusp width, molar gingival width, canine cusp width, canine gingival width, or diastema width. There were slight differences in arch perimeter and midpalatal suture separation. The results of this study showed that the 2-point appliance produced similar effects on the midpalatal suture and the dentition as did the 4-point appliance. It might therefore be considered instead of the 4-point appliance to successfully produce adequate skeletal and dental expansion. PMID- 12637905 TI - Relationship between transverse dental anomalies and skeletal asymmetry. AB - The recognition and elimination of dental compensation is essential in presurgical orthodontic treatment to achieve successful stabilization of the occlusion after surgery. However, the relationship between a transverse dental anomaly and skeletal asymmetry is not fully understood. To evaluate this relationship, frontal cephalometric and 3-dimensional dental model analyses were carried out on 44 adult Japanese Class III patients (mean age 21 years 11 months) who required surgical orthodontic treatment. The patients were divided into 2 groups: a facial asymmetry group in which the mandibular transverse deviation exceeded +/- 1 SD from the norm, and a control group in which the mandibular transverse deviation was within +/- 1 SD of the norm. Statistical comparison with a control group showed characteristic dental anomalies in the facial asymmetry group, including asymmetry of the curve of Spee, molar inclination, dental arch form, lateral overjet, and slanting of the occlusal plane. Stepwise linear regression analysis showed that transverse and vertical skeletal asymmetry variables including the mandible and the maxilla were effective parameters for characteristic dental anomaly variables, and a significant high correlation between dental anomalies and skeletal asymmetry was found. PMID- 12637906 TI - Longitudinal changes in craniofacial factors among snoring and nonsnoring Bolton Brush study participants. AB - The extent to which craniofacial risk factors are manifested from childhood in habitual snorers and those with obstructive sleep apnea hypopnea syndrome (OSAHS) is unknown. The purpose of this study was to examine the differences in craniofacial factors between snorers and nonsnorers from childhood to adulthood. The sample consisted of 80 Bolton-Brush subjects (men, 52%; white, 100%) recalled at adult ages (53-78 years) who had existing prepubertal (age, 4-5 years) and pubertal (age, 12-13 years) records. Snoring was assessed at the adult age through subjective and spousal reports. Cephalometric radiographs were used to characterize 13 craniofacial hard and soft tissue measurements. The t tests revealed that there was a trend (P <.10) for longer hyoid-to-mandibular plane distance at prepubertal and pubertal ages and significant (P <.05) differences at adult recall for snorers. Posterior airway space was significantly smaller for snorers at adult recall. The longitudinal analysis did show a significant change in 9 craniofacial variables over time, but this change was not different between snorers and nonsnorers. We concluded that snorers exhibit a lowered hyoid position from childhood and that longitudinal changes must be explored further with a larger sample. PMID- 12637907 TI - Effect of sustained-release chlorhexidine varnish on Streptococcus mutans and Actinomyces viscosus in orthodontic patients. AB - This study evaluated the effect of sustained-release chlorhexidine varnish on orthodontic patients. Ten children, ages 10 to 16 years, participated. Bacterial levels of Streptococcus mutans and Actinomyces viscosus and total counts were evaluated in sputum samples. These counts were evaluated at 4 stages: before orthodontic treatment, at least 2 weeks after bonding of the brackets, 1 week after application of chlorhexidine varnish, and 3 weeks after application of chlorhexidine varnish. Increases in bacterial levels of S mutans and in the total bacterial count were detected after the brackets were bonded. One week after the sustained-release chlorhexidine varnish was applied, a significant decrease of total bacterial levels and S mutans was observed. This decrease persisted for 3 weeks after the first application. No significant change in A viscosus levels occurred during that period. The results provide additional evidence that sustained-release chlorhexidine varnish decreases S mutans levels in orthodontic patients with fixed appliances and therefore might be useful in preventing caries lesions. PMID- 12637908 TI - Reliability and validity of a computer-based Little irregularity index. PMID- 12637909 TI - Field of dreams. PMID- 12637910 TI - Litigation, legislation, and ethics. Paying it forward: an orthodontist's responsibility for possible future injuries. PMID- 12637912 TI - Cutaneous vasculitis. AB - Vasculitis can range in severity from a self-limited single-organ disorder to a life-threatening disease with the prospect of multiple-organ failure. This condition presents many challenges to the physician, including classification and diagnosis, appropriate laboratory workup, treatment, and the need for careful follow-up. The physician must not only be able to recognize vasculitis but also be able to provide a specific diagnosis (if possible) as well as recognize and treat any underlying etiologic condition. Most diagnostic criteria are based on the size of vessel involvement, which often correlates with specific dermatologic findings. This may allow the dermatologist to provide an initial diagnosis and direct the medical evaluation. This article reviews the classification and diagnosis of cutaneous vasculitic syndromes and current treatment options; it also presents a comprehensive approach to diagnosing and treating the patient with suspected cutaneous vasculitis. (J Am Acad Dermatol 2003;48:311-40.) LEARNING OBJECTIVE: At the completion of this learning activity, participants should be familiar with the classification and clinical features of the various forms of cutaneous vasculitis. They should also have a rational approach to diagnosing and treating a patient with vasculitis. PMID- 12637913 TI - Dermatologic and oral findings in a cohort of 47 patients with Papillon-Lefevre syndrome. AB - Papillon-Lefevre syndrome is an autosomal recessive disorder characterized by palmoplantar hyperkeratosis and early development of aggressive periodontal infection. The aims of this study were to rank the severity of dermatologic and oral affections using a semiquantitative scoring system, and to evaluate whether the severity of the dermatologic changes were correlated to age, degree of periodontal infection, or both. The study included 47 patients with Papillon Lefevre syndrome. With no exception both skin and oral changes developed early in life. The dermatologic involvement showed no correlation with age, whereas the periodontal infection was significantly worse in young children with deciduous teeth. A strong correlation was found between the condition of feet and hands, although the scores for the feet were significantly higher. No significant correlation could be demonstrated between the level of periodontal infection and severity of skin affections, supporting the concept that these 2 major components of Papillon-Lefevre syndrome are unrelated to each other. PMID- 12637914 TI - Comparison of epidermal hydration and skin surface lipids in healthy individuals and in patients with atopic dermatitis. AB - BACKGROUND: The water content of the stratum corneum and skin surface lipids are important factors in the appearance and function of the skin. A disruption of the balance between the two may lead to the clinical manifestation of dryness of skin in patients with atopic dermatitis. OBJECTIVE: The aim of our study was to examine the so-called dry skin of patients with atopic dermatitis using objective parameters. We compared the epidermal hydration and the skin surface lipids, the so-called hydro-lipid film, of the clinically unaffected skin of patients suffering from atopic dermatitis with that of healthy subjects. METHODS: A total of 48 patients of either gender were included in this retrospective case-control study. We used the Corneometer CM 820 (Courage+Khazaka Electronic GmbH, Cologne, Germany) and the Sebumeter SM 810 (Courage+Khazaka Electronic GmbH) as noninvasive measuring methods. RESULTS: The results showed marked decreases in the atopic dermatitis group for both the Corneometer and Sebumeter measuring methods. CONCLUSION: Our results show that the dry skin of patients with atopic dermatitis, as previously shown, is due not only to a decrease in skin moisture but also to a reduction of skin lipids. This finding gives rise to a new understanding of the condition, and therefore one should always speak of a hydro lipid film. PMID- 12637915 TI - Mammographic findings in pseudoxanthoma elasticum. AB - BACKGROUND: There have been isolated case reports of arterial and skin calcification in mammograms of patients with pseudoxanthoma elasticum (PXE), and unpublished anecdotes of many women with PXE undergoing breast biopsy for evaluation of microcalcifications. OBJECTIVE: Our aim was to systematically evaluate mammography and breast pathology in PXE. METHODS: The mammograms of 51 women with confirmed PXE were compared with those of a control sample of 109 women without PXE, noting each of the following characteristics on each mammogram: breast density, skin thickening, skin microcalcifications, vascular calcification, breast microcalcifications and macrocalcifications, and masses. The characteristics of the 2 samples were compared using the 2-tailed t test with a pooled estimate of variance. The indications for mammography and data for each of the mammographic findings were analyzed using the chi(2) test. Available breast biopsy material was reviewed. RESULTS: The PXE and control groups were similar in age and indications for mammography. There was a statistically significant increase in skin thickening, vascular calcification, and breast microcalcifications in the PXE group (P <.001 each). Breast density, masses, macrocalcifications, and skin calcification did not differ statistically in the 2 groups, but no control patient had axillary calcification, or both vascular calcification and microcalcifications (P <.001). Nearly 1 in 7 of the patients with PXE demonstrated at least 3 of the following: microcalcifications, skin calcifications, vascular calcification, and skin thickening; whereas none of the control group did. Histopathologic findings of breast tissue showed calcification of dermal elastic fibers, subcutaneous arteries, and elastic fibers of the deep fascia and interlobular septae of the fat adjacent to breast parenchyma. CONCLUSION: Breast microcalcification and arterial calcification are not rare in the normal population and are not of diagnostic value. The presence of both of these findings, especially with skin thickening or axillary skin calcification, should suggest a diagnosis of PXE. The majority of breast calcifications in PXE are benign. PMID- 12637916 TI - The Eastern Australian Childhood Nevus Study: site differences in density and size of melanocytic nevi in relation to latitude and phenotype. AB - It has been postulated that site-specific variation in melanocytic nevus density and size is explained by differential response to sunlight. We observed the density and size of nevi at different body sites in relation to age, phenotype, latitude, and other measures of ultraviolet exposure. A standard protocol was used to assess nevi, phenotype, and sun exposure in 1123 Australian schoolchildren at 3 contrasting latitudes. Associations with phenotype (red hair, skin reflectance, sun sensitivity, and tanning) varied by body site. In Queensland, gender differences in nevus density on the back and lower limbs, unrelated to sun exposure, were similar to gender differences for melanoma. Small nevi (2-4 mm) were most dense on the arms, whereas large nevi (> or =5 mm) were most dense on the posterior trunk where they were related to age, decreasing latitude, male sex, and freckling. Our findings support the hypothesis of site specific differences in nevus proliferative potential. PMID- 12637917 TI - Interstitial granulomatous dermatitis with histiocytic pseudorosettes: a new histopathologic pattern in cutaneous borreliosis. Detection of Borrelia burgdorferi DNA sequences by a highly sensitive PCR-ELISA. AB - BACKGROUND: The cutaneous manifestations of Borrelia burgdorferi infection include an early phase of erythema chronicum migrans and a late stage of acrodermatitis chronica atrophicans lesions. OBJECTIVE: We describe 11 patients with peculiar cutaneous manifestations and distinctive histopathologic findings as the result of B burgdorferi infection. METHODS: Eleven patients with B burgdorferi detected by polymerase chain reaction or polymerase chain reaction enzyme-linked immunosorbent assay in their cutaneous lesions were included in this study. We analyzed clinical data and histopathologic findings in all patients. The inflammatory infiltrate was also immunohistochemically investigated. RESULTS: Most patients showed a peculiar clinical setting of morphea, and a few cases presented the characteristic appearance of erythema chronicum migrans instead of acrodermatitis chronica atrophicans, as would be expected in a late phase of B burgdorferi infection. The histopathologic findings were similar in all cases and consisted of an interstitial inflammatory infiltrate mostly composed of histiocytes dispersed among the collagen bundles of the dermis and focal areas of small pseudorosette formation, characterized by small histiocytes radially disposed around thick collagen bundles. In some cases there were also a few plasma cells intermingled with the histiocytes. CONCLUSION: Cutaneous lesions with clinical appearance similar to that of morphea and histopathologic features closely resembling those of the interstitial type of granuloma annular may be seen in intermediate-stage cutaneous lesions of B burgdorferi infection. These clinical and histopathologic findings represent a constellation of findings that have not been previously characterized as a cutaneous manifestation of B burgdorferi infection. PMID- 12637918 TI - Human papillomavirus-associated digital squamous cell carcinoma: literature review and report of 21 new cases. AB - OBJECTIVE: Our aim was to review the clinical behavior of human papillomavirus (HPV)-associated digital squamous cell carcinoma (SCC). Specifically, we examined evidence for the tumor's (1) infectious origin and spread, (2) response to therapy, and (3) prognosis and metastatic risk. DESIGN: We reviewed and performed data tabulation of all 51 reported cases in the English-language literature and a case series of 23 cases (21 of them not previously reported). We present 2 of the cases in depth. SETTING: We used previously reported cases from MEDLINE and a case series from a single dermatologic operation practice from 1985 to 1999. RESULTS: (1) Of all cases, 10% (7/72) had an antecedent genital dysplasia or carcinoma containing the same HPV subtype as the digital SCC. (2) The rate of recurrence after general surgical therapy was 43% (6/14). After Mohs micrographic surgery the recurrence rate was 13% (2/16) for the cases in the literature, and 26% (6/23) for our case series. (3) Of tumors, 3% (2/72) have been observed to metastasize. CONCLUSIONS: (1) This suggests the possibility of genital-digital spread as a mechanism of tumor genesis. (2) HPV-associated digital SCC is more likely to recur after surgical treatment than previously reported. This rate of recurrence greatly exceeds that for cutaneous SCCs in general and may be caused by residual postsurgical HPV. (3) The rate of metastasis, however, appears to be low. PMID- 12637920 TI - High frequency and diversity of cutaneous appendageal tumors in organ transplant recipients. AB - BACKGROUND: Recipients of organ transplant who are immunosuppressed are at greatly increased risk of nonmelanoma skin cancers compared with the general population, but their risk of appendageal tumors is unknown. OBJECTIVE: Our aim was to conduct a systematic examination of cutaneous appendageal tumors arising in recipients of organ transplants compared with individuals who were immunocompetent (ICP). METHODS: We conducted a retrospective, clinicopathologic analysis of consecutive appendageal tumors arising in 650 recipients of organ transplants and in the general population of approximately 605,000 people served by our institution. RESULTS: Between 1993 and 1998, 231 appendageal tumors were identified in 211 individuals; 23 tumors were found in 21 of 650 patients undergoing transplant (3%), 10 in individuals with other immunosuppressive conditions, 3 in 2 patients with Muir-Torre syndrome, and 195 in 178 apparently ICP. In addition to the increased frequency of appendageal tumors among recipients of transplants, malignant tumors were overrepresented (43% of transplant tumors vs 4% in ICP; P <.0001) as were tumors of sebaceous origin (30% vs 6%; P <.0001). CONCLUSIONS: Recipients of organ transplant who are immunosuppressed have a greatly increased risk of cutaneous appendageal tumors compared with apparently ICP. In addition, their tumors are more likely to be malignant and of sebaceous origin. PMID- 12637919 TI - Electrical impedance as a potential tool to distinguish between allergic and irritant contact dermatitis. AB - BACKGROUND AND OBJECTIVE: The allergic contact reaction is a model reaction for studying the cell-mediated immune system of the skin. In this study we use a noninvasive method, electrical impedance (IMP), to compare nickel (Ni) allergic contact reactions with an irritant contact reaction induced by sodium lauryl sulfate, which has already been carefully evaluated with this method. For this purpose, we included only Ni- and sodium lauryl sulfate-induced reactions of very similar appearance. METHODS: Various concentrations of Ni sulfate in distilled water were applied on the volar aspect of the forearms of 33 adult women who were allergic to Ni. Assessments were made using visual scoring, a new IMP technique, and transepidermal water loss 3 and 7 days later. RESULTS: In the 19 patients who completed the study, 3 of the 4 impedance indices were significantly lower at the sodium lauryl sulfate sites than at the Ni sites on day 3; ie, the mean magnitude (MIX) (P < or = .001), imaginary part (IMAX) (P < or = .001), and real part (RIX) indices (P < or = .01). Unlike the irritant reactions, no significant increases in transepidermal water loss occurred in the allergic contact reactions. This may be because, in reactions of the studied magnitude, an allergic contact reaction does not significantly affect the epidermis because the inflammatory process is located deeper in the dermis than an irritant reaction. CONCLUSION: This study suggests that IMP, as used herein, is suitable for distinguishing between contact reactions of allergic and irritant nature. Although pathophysiologic events in the tissue studied significantly modify impedance patterns, little is known about how to interpret the structural and chemical changes underlying these patterns. Studies are needed to determine the relation between anatomic or pathophysiologic parameters, and the findings using IMP and other established methods, such as chemical extraction and histopathology. PMID- 12637921 TI - Laboratory tests and identified diagnoses in patients with physical and chronic urticaria and angioedema: A systematic review. AB - BACKGROUND: The value of laboratory tests in chronic urticaria is still controversial. OBJECTIVE: Our aim was to assess this value in clinical studies, and to identify factors explaining the variation in the number of identified causes. METHODS: A total of 4 electronic databases were searched, and a manual literature search was performed. Only unselected patient series with more than 50 adult patients were included. From each included study predefined items were recorded to assess their quality (consecutive patients, use of standardized diagnostic criteria) and validity (follow-up, assessment of treatment effects, level of evidence). RESULTS: A total of 29 studies were included, involving 6462 patients. The verification of the validity of the results and the level of evidence of the included studies were limited. CONCLUSION: No relationship between the number of identified diagnoses and the number of performed laboratory tests, the different settings, the study design, or the publication period was found. On the basis of this systematic review and the relevant literature, a clinical guideline in the form of a flowchart is presented. PMID- 12637922 TI - Experience with low-dose methotrexate for the treatment of eczema in the elderly. AB - There is very little published information on the use of methotrexate in treating eczema. We have used methotrexate in 5 elderly patients with eczema, with a successful response in 4; in 1 patient therapy was stopped as a result of other ongoing medical problems. Methotrexate can be considered as an option in the treatment of eczema unresponsive to topical therapy in elderly patients. PMID- 12637924 TI - Skin cancer is among the most costly of all cancers to treat for the Medicare population. AB - BACKGROUND: Compared with other malignancies, nonmelanoma skin cancer (NMSC) is associated with much less morbidity and mortality. NMSC is, however, far more common than other malignancies. The cost of managing NMSC has not been assessed. OBJECTIVE: The purpose of our study was to determine where the cost of NMSC management ranks among other cancers in the Medicare population. DESIGN: Representative Medicare part A and B claims data were obtained from the Medicare current beneficiary survey, 1992 to 1995. Claims associated with cancer costs were identified using the International Classification of Diseases, Ninth Revision, Clinical Modification codes. Weights were applied to obtain nationally representative estimates. RESULTS: Average Medicare expenditure on cancer management was $13 billion per year. The 5 most costly cancers to Medicare were lung and bronchus, prostate, colon and rectum, breast, and NMSC. The mean annual cost per patient using Medicare for all cancers was $17,094. Malignancies of lung and bronchus, colon and rectum, breast, and prostate were 11 to 19 times more costly per affected patient than NMSC. CONCLUSION: In addition to classifying cancers by number of cases and number of deaths, the financial impact of treatment can also be used to prioritize different malignancies. Such a scheme ranks NMSC far higher than would death statistics. In light of its already high and rising incidence, the cost of NMSC care to Medicare is likely to increase. However, to maintain the cost-effective management of NMSC, it is essential to preserve the current low per-patient cost of its management. PMID- 12637923 TI - Evaluating invasive cutaneous melanoma: is the initial biopsy representative of the final depth? AB - BACKGROUND: An accurate initial biopsy of the deepest portion of the melanoma is vital to the management of patients with melanomas. OBJECTIVE: Our goal was to evaluate the accuracy of preliminary biopsies performed by a group of predominantly experienced dermatologists (n = 46/72). METHODS: A total of 145 cases of cutaneous melanoma were examined retrospectively. We compared Breslow depth on preliminary biopsy with Breslow depth on subsequent excision. Was the initial diagnostic biopsy performed on the deepest part of the melanoma? RESULTS: Of nonexcisional initial shave and punch biopsies, 88% were accurate, with Breslow depth greater than or equal to subsequent excision Breslow depth. Both superficial and deep shave biopsies were more accurate than punch biopsy for melanomas less than 1 mm. Excisional biopsy was found to be the most accurate method of biopsy. CONCLUSIONS: Deep shave biopsy is preferable to superficial shave or punch biopsy for thin and intermediate depth (<2 mm) melanomas when an initial sample is taken for diagnosis instead of complete excision. We found that a group of predominantly experienced dermatologists accurately assessed the depth of invasive melanoma by use of a variety of initial biopsy types. PMID- 12637925 TI - Surgical pearl: autologous biological dressing for epidermal grafting in vitiligo and other achromic disorders. PMID- 12637926 TI - Oral and genital lichenoid reactions associated with circulating autoantibodies to desmoplakins I and II: a novel target antigen or example of epitope spreading? AB - We describe the first reported case of persistent oral and genital lichenoid reactions associated with circulating antibodies to 250- and 215-kd proteins, compatible with desmoplakin I and II, respectively. We discuss the potential role of epitope spreading, leading to the novel development of specific autoantibodies to desmoplakin. PMID- 12637927 TI - Hydroxyurea-induced dermatomyositis-like eruption. AB - Hydroxyurea is a cytotoxic chemotherapeutic agent used for myelodysplasia. The adverse cutaneous effects due to hydroxyurea include leg ulcers, hyperpigmentation of the skin and nails, a lichen planus-like eruption, lupus erythematosus, and a dermatomyositis-like eruption. We present a case of hydroxyurea-induced dermatomyositis-like eruption and review the features of this entity as previously reported. PMID- 12637928 TI - Atypical recurrent varicella in 4 patients with hemopathies. AB - Relapsing varicella may occur in children with HIV infection and more rarely in younger adults. Our aim was to report unusual clinical, histologic, and virologic aspects of 4 elderly patients with malignant hemopathies who had an unusual form of recurrent varicella develop. Conventional microscopy, immunohistochemistry, and in situ hybridization were applied to smears and skin biopsy specimens. The patients presented a few dozen, scattered, large, papulovesicular lesions with central crusting. No zoster-associated pain or dermatomal distribution of the lesions was noted. Conventional microscopy revealed vascular changes and epidermal alterations typical for alpha-herpes virus infection. The varicella zoster virus major viral envelope glycoproteins gE and gB, and the immediate early varicella zoster virus IE63 protein and the corresponding genome sequence for gE were detected on Tzanck smears; they were localized in endothelial cells and keratinocytes on skin biopsy specimens. The varicella zoster virus infection in endothelial cells, the vascular involvement, and the widespread distribution of the lesions suggest that the reported eruptions are vascular rather than neural in origin. These findings invalidate the diagnosis of herpes zoster but strongly support the diagnosis of recurrent varicella in an indolent and yet unreported presentation. Furthermore, these eruptions differ from relapsing varicella in children and young adults by the age of the patients, the paucity of clinical lesions, the larger diameter of the lesions and their peculiar clinical aspect, the significantly longer time interval between primary varicella and the recurrence, the prolonged healing time of the lesions, their mild disease course, and the fact that all the lesions are in the same stage of development. PMID- 12637929 TI - Purely follicular mycosis fungoides without mucinosis: report of two cases with review of the literature. AB - Follicular lesions can either associate with typical patch-/plaque-type mycosis fungoides or, more rarely, be the only clinical manifestation of the disease. We describe 2 adult patients who presented with alopecia and disseminated follicular erythematous papules, and comedones and cysts, respectively. In both patients, histology showed a folliculotropic infiltrate of atypical lymphocytes that spared the epidermis, in the absence of follicular mucinosis. Molecular genetic analysis confirmed the oligo/monoclonal nature of the T-cell infiltrate. Reported cases of purely follicular mycosis fungoides without mucinosis are reviewed. PMID- 12637930 TI - Apoeccrine sweat duct obstruction as a cause for Fox-Fordyce disease. AB - The pathogenesis of Fox-Fordyce disease has been reported to be hyperkeratosis and obstruction of the upper hair follicle, where the duct of the apocrine sweat gland opens. We report a case of Fox-Fordyce disease with full clinical manifestation. It appeared to be caused by the obstruction of intraepidermal apoeccrine sweat ducts by apoeccrine secretory cells detached and released from the secretory epithelium. A 24-year-old woman visited our clinic with intensely pruritic papules on axillae, mammary areolae, and pubic areas. Histopathologic examination revealed an obstruction of the sweat duct in the epidermis, which opened directly to the skin surface. The closing substance of the duct was an aggregate of epithelial cells, probably derived from the secretory portion. In the dermis, the secretory cells of apocrinelike sweat glands had been detaching from the secretory epithelia. These findings suggest that Fox-Fordyce disease can occur by the mechanism in which apoeccrine secretory cells obstruct sweat ducts. PMID- 12637931 TI - Superficial granulomatous pyoderma treated with intravenous immunoglobulin. AB - Superficial granulomatous pyoderma is a rare variant of pyoderma gangrenosum. The superficial ulceration and vegetative margins are clinically distinctive; the suppurative and granulomatous histology is characteristic, though not pathognomonic. The condition is said to be relatively benign and more responsive to treatment than classic pyoderma gangrenosum, though published evidence of this is not always convincing. We present a case of superficial granulomatous pyoderma that was unusually aggressive, but that underwent dramatic and lasting resolution after intravenous immunoglobulin therapy. PMID- 12637932 TI - "Dysplastic nevus" syndrome: does a survey make it real? PMID- 12637933 TI - "Dysplastic nevus" syndrome: does a survey make it real? PMID- 12637934 TI - "Dysplastic nevus" syndrome: does a survey make it real? PMID- 12637935 TI - UVA, melanoma, and antibacterial soaps. PMID- 12637936 TI - Retroviral vector-producer cell-mediated in vivo gene transfer of TIMP-3 restricts angiogenesis and neuroblastoma growth in mice. AB - Destruction and remodeling of the extracellular matrix occurs during the formation of new blood vessels that are required for tumor growth. We sought to determine whether gene-therapy mediated in vivo delivery of tissue inhibitor of matrix metalloproteinase-3 (TIMP-3), using retroviral vector-producer cells, could suppress angiogenesis and subsequent tumor growth in a murine neuroblastoma model. Tumor volume 28 days after coinjection of tumor cells with producer cells generating TIMP-3-encoding retroviral vectors was 21% that of controls, as was the mean tumor vascular index, a measure of blood vessel maturity. When tumors were allowed to reach a mean volume of 0.05 cm(3) before treatment, their size 2 weeks later was 47% relative to controls; larger tumors were not significantly affected. When producer cells were injected at surgical sites following excision of subcutaneous tumors, local recurrence 14 days later was only 22% in TIMP-3 producer cell treated mice as compared to 71% in controls. Unsuccessful transduction of melanoma cells in situ, another tumor of neural crest origin, resulted in unimpaired tumor growth, despite the fact that these tumors are susceptible to TIMP-3 overexpression, demonstrating the importance of tumor cell transduction in this approach. Thus, retroviral vector-producer cell-mediated in vivo gene transfer of TIMP-3 to tumor cells can significantly restrict tumor induced angiogenesis and tumor growth. This approach may be an effective adjuvant in the treatment of neuroblastoma and other solid tumors refractory to traditional therapy, although it appears to be most effective in smaller tumors or in the setting of minimal residual disease, and the tumor cells must be susceptible to retroviral vector-mediated transduction. PMID- 12637937 TI - E2F-1 overexpression sensitizes colorectal cancer cells to camptothecin. AB - Topoisomerase I inhibitors have been shown to have clinical activity against human colorectal cancer. Previous studies showed that the cytotoxicity of camptothecin, a topoisomerase I inhibitor, occurs mainly in the S -phase of the cell cycle and is protectable by aphidicolin, an inhibitor of replicative DNA polymerase in some camptothecin-sensitive colorectal cells. Transcription factor E2F-1 regulates the G1/S transition, and recent studies have shown that E2F-1 potentiated the cytotoxicity of some cell-cycle-related drugs. Therefore, the present study was designed to investigate the effect of adenovirus-mediated E2F-1 gene transfer on chemosensitivity of colorectal cancer to camptothecin, in vitro and in vivo. Two human colorectal cancer cells, SW620 (mutant p53) and RKO (wild type p53), were treated with camptothecin, alone or in combination with adenoviral vectors expressing beta-galactosidase (Ad-LacZ), or E2F-1 (Ad-E2F-1). E2F-1 overexpression was confirmed by Western blot analysis. Ad-E2F-1 gene transfer at low doses (less than the LD(20) dose) markedly increased the sensitivity of human colorectal cancer cells to camptothecin in vitro, which is because of induction of apoptosis. Aphidicolin did not have any protective effect on the Ad-E2F-1/camptothecin-mediated cytotoxicity. The level of topoisomerase I expression was not affected by combination treatment as well, suggesting that DNA replication and topoisomerase I activity may not account for the molecular mechanism of cell killing in response to Ad-E2F-1/camptothecin treatment. Fas and Fas ligand expression were not altered by treatment with camptothecin and/or Ad E2F-1. Moreover, combination of camptothecin and Ad-E2F-1 has an additive antitumor effect in an in vivo nude mouse xenograft model. When combined with camptothecin, E2F-1 adenovirus therapy resulted in a 95.7% decrease in tumor size compared to control groups (P<.05). These results suggest a chemosensitization strategy that may have clinical utility in human colorectal cancer. PMID- 12637938 TI - A novel combination of suicide gene therapy and histone deacetylase inhibitor for treatment of malignant melanoma. AB - One major problem associated with application of gene therapy to treatment of tumors is poor transgene expression. Although suicide gene therapy with the herpes simplex virus-thymidine kinase gene (HSV-tk) followed by administration of ganciclovir (GCV) was effective in the treatment of melanoma, it was still difficult to induce complete remission to cancer. A novel histone deacetylase inhibitor drug FR901229 was found to enhance transgene expression in tumor cells both in vitro and in vivo. Combination therapy with HSV-tklGCV and FR901228 by direct injection into tumor enhanced antimelanoma effects. The number of apoptotic cells in melanoma tumors was increased significantly (P<.05) after combined suicide gene therapy and FR901228. Six times injection of HSV-tk/GCV and FR901228 prolonged mice survival compared to that of HSV-tk/GCV injection alone (P=.021). In total, 56% (10 of 18) of the mice survived 120 days after combined suicide gene therapy and FR901228 treatment, and no new tumors appeared in the surviving mice. However, only 19% (3 of 16) of the mice survived when treated with suicide gene therapy alone. This novel strategy may be applicable as a therapeutic regimen for the treatment of aggressive types of cancers. PMID- 12637939 TI - Expression of the interleukin-21 gene in murine colon carcinoma cells generates systemic immunity in the inoculated hosts. AB - Interleukin-21 (IL-21) is a novel cytokine that can induce proliferation of activated T cells and maturation of natural killer (NK) cells. We therefore examined whether expression of the IL-21 gene in tumor cells could generate antitumor responses. Murine colon carcinoma Colon 26 cells that were transduced with the mouse IL-21 gene (Colon 26/IL-21) were rejected in syngeneic mice and the mice subsequently acquired protective immunity. The growth of Colon 26/IL-21 tumors developed in nude mice was retarded compared with that of parent tumors, and this growth suppression was not observed in nude mice that were treated with anti-asialo GM(1) antibody. Spleen cells from the mice that had rejected Colon 26/IL-21 cells showed cytotoxic activity to Colon 26 but not to irrelevant tumor cells, and produced larger amounts of interferon-gamma upon stimulation with irradiated Colon 26 cells. Spleen cells from Colon 26/IL-21-tumor- but not parent tumor-bearing mice had lytic activity to YAC-1 cells. These data suggest that expression of IL-21 in tumors induces T- and NK-cell-dependent antitumor effects. PMID- 12637940 TI - Radiation increases the activity of oncolytic adenovirus cancer gene therapy vectors that overexpress the ADP (E3-11.6K) protein. AB - We have described three potential adenovirus type 5 (Ad5)-based replication competent cancer gene therapy vectors named KD1, KD3, and VRX-007. All three vectors overexpress an Ad5 protein named Adenovirus Death Protein (ADP, also named E3-11.6 K protein). ADP is required for efficient lysis of Ad5-infected cells and spread of virus from cell to cell, and thus its overexpression increases the oncolytic activity of the vectors. KD1 and KD3 contain mutations in the Ad5 E1A gene that knock out binding of the E1A proteins to cellular p300/CBP and pRB; these mutations allow KD1 and KD3 to grow well in cancer cells but not in normal cells. VRX-007 has wild-type E1A. Here we report that radiation increases the oncolytic activity of KD1, KD3, and VRX-007. This increased activity was observed in cultured cells, and it was not because of radiation induced replication of the vectors. The combination of radiation plus KD3 suppressed the growth of A549 lung adenocarcinoma xenografts in nude mice more efficiently than radiation alone or KD3 alone. The combination of ADP overexpressing vectors and radiation may have potential in treating cancer. PMID- 12637941 TI - Downregulation of bcl-2 expression in lymphoma cells by bcl-2 ARE-targeted modified, synthetic ribozyme. AB - Synthetic ribozymes are catalytic RNA molecules designed to inhibit gene expression by cleaving specific mRNA sequences. We investigated the potential of synthetic ribozymes to inhibit bcl-2 expression in apoptosis defective bcl-2 overexpressing tumors. A chemically stabilized hammerhead ribozyme has been targeted to the A+U-rich regulative element of bcl-2 mRNA that is involved in bcl 2 gene switch-off during apoptosis. The design of the ribozyme was based on the results of probing accessibility of the RNA target in cellular extracts with antisense DNA. The ribozyme was lipotransfected to a bcl-2 overexpressing human lymphoma cell line (Raji). The cellular uptake of this ribozyme resulted in a marked reduction of both bcl-2 mRNA and BCL-2 protein levels and dramatically increased cellular death by apoptosis. Our results suggest a potential therapeutic application of such ribozyme for the treatment of bcl-2 overexpressing tumors. PMID- 12637942 TI - Dendritic cells transfected with tumor RNA for the induction of antitumor CTL in colorectal cancer. AB - Dendritic cells (DC) are the most potent antigen-presenting cells known, currently tested for vaccination studies in cancer patients. The use of tumor derived RNA to load DC overcomes the requirement of defined HLA types and the identification of tumor antigens expressed by the tumors. Here, we show that human monocyte-derived DC generated under serum-free conditions by GM-CSF, IL-4 and TNF-alpha acquire a mature phenotype and expression of the chemokine receptor CCR-7, which plays a pivotal role in DC migration to the afferent lymph nodes. We demonstrate the feasibility of total RNA transfection into such DC using the renal cell carcinoma (RCC) cell line N43-EGFP, which was stably transfected with an EGFP-encoding vector. Moreover, we show that DC transfected with RNA from colorectal cancer cells present HLA class I-restricted antigenic epitopes to induce a primary antitumor CTL response in vitro. Interestingly, the CTL induced by SW480 RNA also recognized another colon cancer line, HCT116, and the RCC line A498. Our results confirm the feasibility of total RNA transfection of serum-free generated DC for the induction of CTL against colon cancer and RCC cells, and support the relevance of shared tumor rejection epitopes between colorectal cancer and RCC. PMID- 12637943 TI - Neoadjuvant treatment of hepatic malignancy: an oncolytic herpes simplex virus expressing IL-12 effectively treats the parent tumor and protects against recurrence-after resection. AB - The objective of the study was to evaluate the utility of NV1042, a replication competent, oncolytic herpes simplex virus (HSV) containing the interleukin-12 (IL 12) gene, as primary treatment for hepatic tumors and to further assess its ability to reduce tumor recurrence following resection. Resection is the most effective therapy for hepatic malignancies, but is not possible in the majority of the patients. Furthermore, recurrence is common after resection, most often in the remnant liver and likely because of microscopic residual disease in the setting of postoperative host cellular immune dysfunction. We hypothesize that, unlike other gene transfer approaches, direct injection of liver tumors with replication competent, oncolytic HSV expressing IL-12 will not only provide effective control of the parent tumor, but will also elicit an immune response directed at residual tumor cells, thus decreasing the risk of cancer recurrence after resection. Solitary Morris hepatomas, established in Buffalo rat livers, were injected directly with 10(7) particles of NV1042, NV1023, an oncolytic HSV identical to NV1042 but without the IL-12 gene, or with saline. Following tumor injection, the parent tumors were resected and measured and the animals were challenged with an intraportal injection of 10(5) tumor cells, recreating the clinical scenario of residual microscopic cancer. In vitro cytotoxicity against Morris hepatoma cells was similar for both viruses at a multiplicity of infection of 1 (MOI, ratio of viral particles to target cells), with >90% tumor cell kill by day 6. NV1042 induced high-level expression of IL-12 in vitro, peaking after 4 days in culture. Furthermore, a single intratumoral injection of NV1042, but not NV1023, induced marked IL-12 and interferon-gamma (IFN-gamma) expression. Both viruses induced a significant local immune response as evidenced by an increase in the number of intratumoral CD4(+) and CD8(+) lymphocytes, although the peak of CD8(+) infiltration was later with NV1042 compared with NV1023. NV1042 and NV1023 reduced parent tumor volume by 74% (P<.003) and 52% (P<.03), respectively, compared to control animals. Treatment of established tumors with NV1042, but not with NV1023, significantly reduced the number of hepatic tumors after resection of the parent tumor and rechallenge (16.8+/-11 (median=4) vs. 65.9+/-15 (median=66) in control animals, P<.025). In conclusion, oncolytic HSV therapy combined with local immune stimulation with IL-12 offers effective control of parent hepatic tumors and also protects against microscopic residual disease after resection. The ease of use of this combined modality approach, which appears to be superior to either approach alone, suggests that it may have clinical relevance, both as primary treatment for patients with unresectable tumors and also as a neoadjuvant strategy for reducing recurrence after resection. PMID- 12637945 TI - Dendritic cells reconstituted with human telomerase gene induce potent cytotoxic T-cell response against different types of tumors. AB - Human telomerase reverse transcriptase (hTERT) is the catalytic component of a functional telomerase complex, which is important in maintaining cell immortality. In most normal human adult cells, the expression of telomerase is very low and/or transient. In contrast, almost 90% of human tumors express a relatively high level of telomerase implying the possibility of using hTERT as a universal candidate tumor antigen. In this study, we show that human monocyte derived dendritic cells (DCs) lack telomerase activity. Similar to other normal somatic cells, DCs express the RNA (hTR) component but not the catalytic component, hTERT. We also show that telomerase activity could be reconstituted using either lipid-mediated transfection of the hTERT plasmid DNA or transduction with an E1-, E3-deleted adenoviral vector containing the hTERT gene. However, relative to plasmid transfection, adenoviral gene transfer produced higher levels of hTERT expression. Nine of 10 AdhTERT-transduced DCs were able to generate CTL responses, while only three of nine plasmid-transfected DCs did. CTLs primed against hTERT exhibited killing of telomerase positive but not telomerase negative tumor lines of diverse tissue origins. Antigenic specificity of these T cells to telomerase was further determined by introducing hTERT gene into a telomerase negative cell line, U2OS, by adenoviral transduction. Although some antigenic specificity was directed against adenoviral epitopes, the majority of CTLs were targeted against telomerase-derived antigen(s). Thus, the hTERT gene, particularly as delivered via the recombinant adenovirus, may be useful as vaccine to induce specific T-cell-mediated tumor immunity in cancer patients. In addition, our results suggest that telomerase activity and/or telomerase expression after hTERT gene transfer have a predictive value in the success of hTERT/DC-based cancer vaccination. PMID- 12637944 TI - Assessment of p53 gene transfer and biological activities in a clinical study of adenovirus-p53 gene therapy for recurrent ovarian cancer. AB - A cohort study was designed to evaluate the efficiency of gene transfer and whether biological activity from the expressed therapeutic gene resulted after administration of a recombinant adenovirus containing the human wild-type p53 (p53(wt)) gene (rAd-p53 SCH 58500). The cohort study was conducted in five trial subjects with recurrent ovarian cancer. Each trial subject received multiple cycles of rAd-p53 SCH 58500, each cycle comprised of doses of 7.5 x 10(13) particles on each of five consecutive days. Subjects were treated with rAd-p53 SCH 58500 alone during Cycle 1 and in combination with gemcitabine during the subsequent cycles. Both tumor biopsies and peritoneal aspirates were collected and evaluated for gene transfer and evidence of the biological activities of the expressed p53(wt) gene. Using quantitative PCR and RT-PCR, and in situ PCR, gene transfer and expression were documented in tumor biopsies (four of five patients) collected from Cycle 1. Furthermore, upregulation of p21/WAF1, bax and mdm-2, and downregulation of survivin were observed in these same tumor biopsy samples, suggesting that intraperitoneal administration of rAd-p53 SCH 58500 leads to detectable p53 biological activity in target tumor tissue. In addition, gene transfer and its expression were observed in cells obtained from peritoneal aspirates. These fluids were mainly comprised of polymorphonuclear neutrophils, indicating that successful gene transfer can be achieved by multiple cycle intraperitoneal administration of recombinant adenovirus. PMID- 12637946 TI - Alcohol stimulates the release of dopamine in the ventral pallidum but not in the globus pallidus: a dual-probe microdialysis study. AB - The mesoaccumbens dopamine system has been hypothesized to be a common neural substrate mediating the actions of various drugs of abuse, including ethanol. However, the involvement of the mesopallidal dopamine system has received very little attention. The present study examined the effects of intraperitoneal (IP) ethanol administration on the extracellular levels of dopamine in the ventral pallidum (VP) and globus pallidus (GP) of Wistar rats. Rats were bilaterally implanted with microdialysis probes aimed at the VP and GP or nucleus accumbens (NAc) and dorsal striatum (dSTR). During microdialysis testing, rats with probes located in the VP and GP were injected IP with sterile saline or 15% (v/v) ethanol in saline at doses of 0.75, 1.5, or 2.25 g/kg. Rats with NAc and dSTR probes were injected with saline or 2.25 g/kg ethanol. The IP administration of 1.5 and 2.25 g/kg ethanol significantly (p <0.05) elevated the extracellular levels of dopamine in the VP (maximal increase: 136 and 182% of baseline, respectively) but not in the GP. No effects on extracellular dopamine levels were observed following the IP injections of 0.75 g/kg ethanol or saline. The IP administration of 2.25 g/kg ethanol significantly (p <0.05) elevated the extracellular levels of dopamine in the NAc (maximal increase: 198% of baseline) and dSTR (maximal increase: 155% of baseline). Analysis of the effects of 2.25 g/kg ethanol on dopamine release revealed greater increases in the VP, NAc, and dSTR compared to the GP. The data suggest that the mesopallidal, mesoaccumbens, and nigrostriatal dopamine systems are more sensitive to the effects of ethanol than the nigropallidal dopamine system. PMID- 12637947 TI - Downregulation of neuronal cdk5/p35 in opioid addicts and opiate-treated rats: relation to neurofilament phosphorylation. AB - Neuronal cyclin-dependent kinase-5 (Cdk5) and its neuron-specific activator p35 play a major role in regulating the cytoskeleton dynamics. Since opioid addiction was associated with hyperphosphorylation of neurofilament (NF) in postmortem human brains, this study was undertaken to assess the status of the cdk5/p35 complex and its relation with NF-H phosphorylation in brains of chronic opioid abusers. Decreased immunodensities of cdk5 (18%) and p35 (26-44%) were found in the prefrontal cortex of opioid addicts compared with matched controls. In the same brains, the densities of p25 (a truncated neurotoxic form of p35), phosphatase PP2Ac and mu-calpain were found unaltered. Acute treatment of rats with morphine (30 mg/kg, 2 h) increased the density of cdk5 (35%), but not that of p35, in the cerebral cortex. In contrast, chronic morphine (10-100 mg/kg for 5 days) induced marked decreases in cdk5 (40%) and p35 (47%) in rat brain. In brains of opioid addicts, the density of phosphorylated NF-H was increased (43%) as well as the ratio of phosphorylated to nonphosphorylated NF-H forms (two fold). In these brains, phosphorylated NF-H significantly correlated with p35 (r=0.58) but not with cdk5 (r=0.03). The results suggest that opiate addiction is associated with downregulation of cdk5/p35 levels in the brain. This downregulation and the aberrant hyperphosphorylation of NF-H proteins might have important consequences in the development of neural plasticity associated with opiate addiction in humans. PMID- 12637948 TI - Rapid assessment of choice between cocaine and food in rhesus monkeys: effects of environmental manipulations and treatment with d-amphetamine and flupenthixol. AB - The present study describes a procedure that permits rapid assessment of environmental and pharmacological factors that may influence the choice between cocaine and food in rhesus monkeys. Daily 2 h sessions were divided into five components. During each component, monkeys (N=4) chose between i.v. cocaine (0 0.1 mg/kg/injection) and food (0, 1, or 3 food pellets). Up to 10 reinforcers were available during each component, and different discriminative stimuli were associated with each magnitude of each reinforcer. Cocaine choice was directly related to cocaine dose, and a cocaine choice dose-effect curve could be determined in a single experimental session. The choice between cocaine and food was influenced by the schedules of cocaine and food reinforcement, the magnitude of the food reinforcer, and the amount of noncontingent food provided outside the experimental session. These results confirm and extend previous findings with other choice procedures and validate the sensitivity of the present procedure to environmental manipulations. The choice between cocaine and food could also be influenced by treatment with candidate pharmacotherapies for cocaine abuse and dependence. The "agonist" medication d-amphetamine produced rightward shifts in the cocaine choice dose-effect curve and decreased cocaine choice, whereas the "antagonist" medication flupenthixol had little effect on cocaine choice. Overall, these results suggest that this choice procedure may be useful for the evaluation of both environmental determinants of cocaine use and candidate pharmacotherapies for the treatment of cocaine abuse. PMID- 12637949 TI - Antidepressant-like effect of different estrogenic compounds in the forced swimming test. AB - The present study evaluated the possible antidepressant-like action of the natural estrogen 17beta-estradiol (E(2), 2.5-10 microg/rat), the synthetic steroidal estrogen ethinyl-estradiol (EE(2), 1.25-10.0 microg/rat), and the nonsteroidal synthetic estrogen, diethyl-stilbestrol (DES, 0.25-1.0 mg/rat) in ovariectomized adult female Wistar rats using the forced swimming test (FST). The behavioral profile induced by the estrogens was compared with that induced by the antidepressants fluoxetine (FLX, 2.5-10 mg/kg) and desipramine (DMI, 2.5-10 mg/kg). In addition, the temporal course of the antidepressant-like action of the estrogenic compounds was analyzed. FLX and DMI induced an antidepressant-like effect characterized by a reduced immobility and increased swimming for FLX and decreased immobility and increased climbing for DMI. Both E(2) and EE(2) produced a decrease in immobility and an increase in swimming, suggesting an antidepressant-like action. DES did not affect the responses in this animal model of depression at any dose tested. The time course analysis of the actions of E(2) (10 microg/rat) and EE(2) (5 microg/rat) showed that both compounds induced an antidepressant-like effect observed 1 h after their injection lasting for 2-3 days. PMID- 12637950 TI - Decreased cocaine self-administration in Kir3 potassium channel subunit knockout mice. AB - Multiple G protein-linked neurotransmitter systems have been implicated in the behavioral effects of cocaine. While actions of certain neurotransmitter receptor subtypes and transporters have been identified, the role of individual G protein regulated enzymes and ion channels in the effects of cocaine remains unclear. Here, we assessed the contribution of G protein-gated, inwardly rectifying potassium (Kir3/GIRK) channels to the locomotor-stimulatory and reinforcing effects of cocaine using knockout mice lacking one or both of the key neuronal channel subunits, Kir3.2 and Kir3.3. Cocaine-stimulated increases in horizontal locomotor activity in wild-type, Kir3.2 knockout, Kir3.3 knockout, and Kir3.2/3.3 double knockout mice, with only minor differences observed between the mouse lines. In contrast, Kir3.2 and Kir3.3 knockout mice exhibited dramatically reduced intravenous self-administration of cocaine relative to wild-type mice over a range of cocaine doses. Paradoxically, Kir3.2/3.3 double knockout mice self-administered cocaine at levels significantly higher than either single knockout alone. These findings suggest that Kir3 channels play significant and complex roles in the reinforcing effect of cocaine. PMID- 12637951 TI - Reversal of a vigilance decrement in the aged rat by subtype-selective nicotinic ligands. AB - In humans, nicotine has been demonstrated to improve both normal and disordered attention, suggesting potential clinical utility for nicotinic ligands. However, attempts to replicate these findings in the rodent have met with some difficulty, thus hampering the search for specific receptor mechanisms underlying these effects. In the present studies, we sought to characterize the effects of nicotine and subtype-selective ligands in a group of aged rats, which show consistent deficits in sustained attention over prolonged sessions of responding in the five-choice serial reaction time task (5-CSRTT). Following the establishment of a replicable performance improvement with nicotine (0.4 mg/kg), we assessed the effects of both SIB 1765F (1-5 mg/kg) and AR-R17779 (20 mg/kg), agonist ligands with selective affinities for the alpha(4)beta(2) and alpha(7) receptor sites, respectively. We then attempted to block this effect of nicotine using the high affinity, competitive nicotinic antagonist DHbetaE (3 mg/kg). Finally, in an attempt to determine whether the psychostimulant profile of nicotinic agonists could be dissociated from their effects on attention, we compared the (R)- and (S)-enantiomers of SIB 1765F in the 5-CSRTT, and in their ability to increase locomotor activity. Reversal of a within-session decline in performance speed and accuracy by nicotine was mimicked by SIB 1765F, but not by AR-R17779, whereas DHbetaE antagonized all of the performance changes induced by nicotine. Finally, the (S)- but not the (R)-enantiomer increased locomotor activity and improved performance in the 5-CSRTT. These results support a critical involvement for the alpha(4)beta(2) nicotinic receptor in mediating the attention-enhancing properties of nicotine. PMID- 12637952 TI - Flattening the glucocorticoid rhythm causes changes in hippocampal expression of messenger RNAs coding structural and functional proteins: implications for aging and depression. AB - Subtle changes in glucocorticoid levels, including a flattening of the diurnal rhythm with raised nadir, are prevalent, being characteristic of both aging and major depression. Both these conditions are also associated with deficits in hippocampally mediated cognitive functions. We hypothesized that this profile of glucocorticoid levels causes structural and functional changes in the hippocampus, which in turn may engender cognitive deficits. We implanted slow release corticosterone pellets into adrenally intact adult male rats to produce a flattened glucocorticoid rhythm with levels clamped midway between the normal nadir and zenith. Using density profile analysis we measured hippocampal expression of messenger RNAs encoding structural and functional proteins. In rats with a flattened glucocorticoid rhythm, the expression of the mRNA coding for microtubule associated protein-2b (MAP2b) was reduced in CA3 relative to sham operated controls, but unchanged in dentate gyrus and CA1. In contrast, the expression of the mRNA coding the alpha subunit of calcium-calmodulin dependent kinase (CAMKIIalpha) was reduced in dentate gyrus in animals with a flattened glucocorticoid rhythm, but unchanged in CA3. The expression of the mRNA coding the synaptic vesicle protein synaptophysin was unchanged in both CA3 and dentate gyrus. The data indicate that a flattening of the normal diurnal glucocorticoid rhythm decreases the hippocampal expression of mRNAs coding key structural and functional proteins, and does so in a regionally selective manner. The data may have relevance for cognitive deficits characteristic of aging and depression. PMID- 12637953 TI - Effects of mixed-action kappa/mu opioids on cocaine self-administration and cocaine discrimination by rhesus monkeys. AB - kappa-Opioid agonists may functionally antagonize some behavioral effects of cocaine, but the role of mixed kappa/mu receptor activity is unclear. The effects of three mixed kappa/mu agonists (MCL-101, (-)cyclorphan, and Mr2034) and one kappa-selective agonist (enadoline) on cocaine self-administration and cocaine discrimination were compared in rhesus monkeys. Acute treatment with all kappa agonists dose dependently reduced cocaine-maintained responding and produced a downward shift in the cocaine self-administration dose-effect curve (0.001-0.32 mg/kg/inj, i.v.). During 7 days of chronic treatment, (-)cyclorphan (0.0032-0.032 mg/kg/h) and MCL-101 (0.0032-0.032 mg/kg/h) each dose dependently reduced cocaine self-administration maintained by a dose near the peak of the cocaine self administration dose-effect curve. MCL-101 (0.032 mg/kg/h) produced selective and sustained decreases in cocaine self-administration, whereas (-)cyclorphan (0.032 mg/kg/h) had selective but transient effects. In addition, these mixed kappa/mu agonists produced fewer side effects (some salivation) than the kappa-selective agonist (sedation, salivation, emesis). However, none of these kappa agonists substituted for or antagonized cocaine's discriminative stimulus effects in monkeys trained to discriminate cocaine (0.4 mg/kg, i.m.) from saline. Thus, kappa and mixed kappa/mu-opioid agonists may reduce cocaine self-administration without altering cocaine's discriminative stimulus effects. Mixed kappa/mu agonists appear to offer some advantages over selective kappa agonists as potential treatments for cocaine abuse. PMID- 12637954 TI - 5-HT 1A/1B receptor-mediated effects of the selective serotonin reuptake inhibitor, citalopram, on sleep: studies in 5-HT 1A and 5-HT 1B knockout mice. AB - Selective serotonin reuptake inhibitors (SSRIs) are extensively used for the treatment of depression. Aside from their antidepressant properties, they provoke a deficit in paradoxical sleep (PS) that is most probably mediated by the transporter blockade-induced increase in serotonin concentration in the extracellular space. Such an effect can be accounted for by the action of serotonin at various types of serotonergic receptors involved in PS regulation, among which the 5-HT(1A) and 5-HT(1B) types are the best candidates. According to this hypothesis, we examined the effects of citalopram, the most selective SSRI available to date, on sleep in the mouse after inactivation of 5-HT(1A) or 5 HT(1B) receptors, either by homologous recombination of their encoding genes, or pharmacological blockade with selective antagonists. For this purpose, sleep parameters of knockout mice that do not express these receptors and their wild type counterparts were monitored during 8 h after injection of citalopram alone or in association with 5-HT(1A) or 5-HT(1B) receptor antagonists. Citalopram induced mainly a dose-dependent inhibition of PS during 2-6 h after injection, which was observed in wild-type and 5-HT(1B)-/- mice, but not in 5-HT(1A)-/- mutants. This PS inhibition was fully antagonized by pretreatment with the 5 HT(1A) antagonist WAY 100635, but only partially with the 5-HT(1B) antagonist GR 127935. These data indicate that the action of the SSRI citalopram on sleep in the mouse is essentially mediated by 5-HT(1A) receptors. Such a mechanism of action provides further support to the clinical strategy of antidepressant augmentation by 5-HT(1A) antagonists, because the latter would also counteract the direct sleep-inhibitory side-effects of SSRIs. PMID- 12637955 TI - Post-mortem interval effects on the phosphorylation of signaling proteins. AB - Post-mortem brain tissue provides a unique opportunity to uncover the genes or proteins involved in the pathophysiology of neuropsychiatric disorders. Protein phosphorylation is a common protein modification within intracellular signaling pathways that affects the distribution and function of protein, and has been hypothesized to be of major importance in both the pathophysiology and treatment of major neuropsychiatric disorders. Thus, we were interested in ascertaining the stability of the phosphorylated forms of proteins that are involved in cellular signaling. Antibodies against phospho-tyrosine, phospho-threonine, and phospho PKA substrates were used to examine the PMI effects on the general amounts of proteins in their phosphorylated form. Phospho-specific antibodies for ERK, JNK, RSK, CREB, and ATF-2 were used to test the effects of PMI on specific proteins whose functioning are known to be regulated markedly by phosphorylation. We found that PMI rapidly decreased the levels of proteins in their phosphorylated states and also decreased the total levels of certain proteins. The PMI effects were observed in the samples stored at both 4 degrees C and room temperature, in both frontal cortex and hippocampus. Thus, it appears that measurements (such as two dimensional gel electrophoresis and functional assays) that rely on the phosphorylation state of proteins would be extremely sensitive to PMI. PMID- 12637957 TI - Norepinephrine but not serotonin reuptake inhibitors enhance theta and gamma activity of the septo-hippocampal system. AB - Current neurobiological concepts attribute a central role of the hippocampal formation in cognitive and affective processes. Recent studies indicate that the hippocampus is affected in human depression, and antidepressant drugs induce hippocampal adaptive changes that are thought to be associated with their therapeutic action. In the present study, we investigated the action of various antidepressant drugs on the activity of the septo-hippocampal system, its oscillatory activity in particular. The acute effects of the norepinephrine (NE) reuptake inhibitors reboxetine and desipramine, and the selective serotonin reuptake inhibitor fluvoxamine were evaluated. Extracellular single-unit recordings were performed from the medial septum/diagonal band of Broca (MS/DBv), with simultaneous hippocampal EEG recordings of anesthetized rats. Systemic administration of reboxetine synchronized hippocampal EEG, resulting in a significant increase in power at theta frequency, and an increase in frequency and power of gamma-wave activity. Parallel to EEG synchrony, reboxetine induced or enhanced theta oscillation of MS/DBv neurons. Oscillatory frequencies of MS/DBv neurons were identical, and phase locked to the corresponding hippocamapal theta frequencies. Under the same experimental conditions, reboxetine induced a two-fold increase in extracellular NE (but not serotonin) levels in the hippocampus as revealed by microdialysis. Desipramine, but not the serotonin reuptake inhibitor fluvoxamine, evoked responses similar to those of reboxetine regarding septo-hippocampal theta activity. The present findings indicate that even though both NE and serotonin reuptake inhibitors are clinically effective antidepressant drugs, their action on the septo-hippocampal oscillatory behavior is different. It is presumed that selective NE reuptake inhibitors could modulate various cognitive processes associated with hippocampal oscillatory activity. PMID- 12637956 TI - Selective antagonism at dopamine D3 receptors enhances monoaminergic and cholinergic neurotransmission in the rat anterior cingulate cortex. AB - Recent neuroanatomical and functional investigations focusing on dopamine (DA) D(3) receptors have suggested a potential role of this receptor in psychiatric diseases such as schizophrenia and drug dependence. In line with the key role of the prefrontal cortex in psychiatric disorders, the present study aimed at assessing the effects of the acute systemic administration of the selective DA D(3) receptor antagonist SB-277011-A on the in vivo extracellular levels of monoamines (DA, norepinephrine (NE), and serotonin (5-HT)) and acetylcholine (ACh) in the anterior cingulate subregion of the medial prefrontal cortex. The in vivo neurochemical profile of SB-277011-A (10 mg/kg, i.p.) in the anterior cingulate cortex was compared with both typical and atypical antipsychotics including clozapine (10 mg/kg, s.c.), olanzapine (10 mg/kg, s.c.), sulpiride (10 mg/kg, s.c.), and haloperidol (0.5 mg/kg, s.c.). The acute administration of SB 277011-A, clozapine, and olanzapine produced a significant increase in extracellular levels of DA, NE, and ACh without affecting levels of 5-HT. Sulpiride also significantly increased extracellular DA, but with a delayed onset over SB-277011-A, clozapine, and olanzapine. In contrast, haloperidol failed to alter any of the three monoamines and ACh in the anterior cingulate cortex. These findings add to a growing body of evidence suggesting a differentiation between typical and atypical antipsychotic drugs (APDs) in the anterior cingulate cortex and a role of DA D(3) receptors in desired antipsychotic drug profile. Similar to their effects on DA and NE, SB-277011-A, clozapine, and olanzapine increased extracellular levels of ACh, whereas haloperidol and sulpiride did not alter ACh. The results obtained in the present study provide evidence of the important role of DA D(3) receptors in the effect of pharmacotherapeutic agents that are used for the treatment of psychiatric disorders such as schizophrenia and drug dependence. PMID- 12637959 TI - NTP Toxicology and Carcinogenesis Studies of Methyl Bromide (CAS: 74-83-9) in B6C3F1 Mice (Inhalation Studies). AB - Methyl bromide is widely used as a fumigant and pesticide. Toxicology and carcinogenesis studies were conducted by exposing groups of male and female B6C3F1 mice to methyl bromide (99.8% pure) by inhalation 6 hours per day, 5 days per week, for 14 days, 6 weeks, 13 weeks, or 2 years. Six-week and 13-week inhalation toxicity studies in F344/N rats were conducted concurrently with the mouse studies. Hematology parameters were measured during the 6-week, 13-week, and 2-year studies. Quantitative neurobehavioral testing was performed during the 14-day, 13-week and 2-year studies. Genetic toxicology studies were conducted for gene mutation induction in Salmonella typhimurium and for induction of sister chromatid exchanges in mouse bone marrow cells and of micronuclei from peripheral blood erythrocytes. 14-Day Studies: Groups of five B6C3F1 mice of each sex were exposed to 0, 12, 25, 50, 100, or 200 ppm methyl bromide by inhalation 6 hours per day, 5 days per week for 2 weeks. Only four female mice and one male mouse survived 10 exposures at 200 ppm. No deaths occurred at the lower doses. Neurobehavioral effects including trembling and paralysis were noted in all groups, but were most pronounced in the three highest dose groups. Red urine was noted in the mice exposed to 200 ppm. 13-Week Studies : Groups of 10 mice of each sex were exposed to 0, 10, 20, 40, 80, or 120 ppm methyl bromide by inhalation 6 hours per day, 5 days per week for 13 weeks. Additional groups of eight to 17 mice were concurrently exposed for neurobehavioral and genetic toxicology studies. The final mean body weight of males exposed to 120 ppm was significantly (12%) lower than that of the controls. Four of 24 males exposed to 120 ppm died during the study. Groups of 10 rats of each sex were exposed to 0, 30, 60, or 120 ppm methyl bromide by inhalation 6 hours per day, 5 days per week for 13 weeks. Additional groups of eight rats were concurrently exposed for neurobehavioral studies. Final mean body weights of rats exposed to 120 ppm were 12% lower than those of the controls for males and 13% lower for females. No rats died as a result of methyl bromide exposure during the studies. Special 6-Week Target Organ Toxicity Studies: Neither the 14-day nor the 13-week studies provided strong evidence for specific organ toxicity. Six-week studies were therefore conducted to identify target organs for the 2-year studies. Groups of 20 rats and mice of each sex were exposed to methyl bromide by inhalation for 6 hours per day, 5 days per week for 6 weeks at a dose of 160 ppm. Mortality rates exceeded 50% in the male mice after eight exposures, in female mice after six exposures, and in male rats after 14 exposures. Only the female rat group survived 30 exposures with less than 50% mortality. The study identified the brain, kidney, nasal cavity, heart, adrenal gland, liver, and testis as the primary organs to examine for toxicity in the 2-year methyl bromide inhalation studies. 2-Year Studies: Groups of 70 B6C3F1 mice of each sex were exposed to methyl bromide by inhalation at 0, 10, 33, or 100 ppm for 6 hours per day, 5 days per week for up to 103 weeks. Additional groups of 16 mice were included for neurobehavioral evaluations throughout the 2-year studies. By 20 weeks (139 days), 27 males and 7 females exposed to 100 ppm had died and methyl bromide exposure was discontinued for the remaining mice in this dose group. Ten female mice from the 100 ppm group predesignated for the 15-month interim evaluation were killed on schedule and all other high-dose animals were allowed to live to term (24 months) for evaluation of chronic toxicity and carcinogenicity. Clinical signs indicative of neurotoxicity, including tremors, abnormal posture, tachypnea, and hind leg paralysis, persisted in these high-dose mice until the end of the studies. Final mean body weights of surviving 100 ppm males and females were markedly lower (33% and 31%) than those of the controls. Neurobehavioral changes occurred in male and female mice initially exposed to 100 ppm methyl bromide, with more prnitially exposed to 100 ppm methyl bromide, with more pronounced changes observed in males. In general, these animals were less active and manifested a heightened sensitivity in the startle response than mice in other dose groups. Exposure to methyl bromide was not carcinogenic under the conditions of these studies. However, there was an increase in the incidence of several nonneoplastic lesions in the brain, heart, bone (sternum), and nose. Degenerative changes in the cerebellum and cerebrum occurred in males and females exposed to 100 ppm. Myocardial degeneration and cardiomyopathy were observed in the hearts of mice exposed to 100 ppm. An increased incidence of sternal dysplasia was seen in treated animals, particularly in those exposed to 100 ppm. An increased incidence of olfactory epithelial necrosis and metaplasia within the nasal cavity was seen in the mice exposed to 100 ppm, particularly males. Genetic Toxicology: Methyl bromide was positive for induction of gene mutations in Salmonella typhimuriumstrain TA100, with and without exogenous metabolic activation; negative results were obtained with TA98 in this assay. In vivo, methyl bromide induced sister chromatid exchanges in bone marrow cells and micronuclei in peripheral erythrocytes of female mice exposed by inhalation for 14 days. No significant increase in either sister chromatid exchanges or micronuclei was observed in male or female mice exposed to methyl bromide by inhalation for 4, 8, or 12 weeks. Conclusions: Under the conditions of these 2-year inhalation studies, methyl bromide caused degenerative changes in the cerebellum and cerebrum, myocardial degeneration and cardiomyopathy, sternal dysplasia, and olfactory epithelial necrosis and metaplasia. Toxic effects persisted although exposure to methyl bromide in the 100 ppm group terminated after 20 weeks. There was no evidence of carcinogenic activity of methyl bromide in male or female B6C3F1 mice exposed to 10, 33, or 100 ppm. Synonym: Bromomethane PMID- 12637960 TI - NTP Toxicology and Carcinogenesis Studies of Titanocene Dichloride (CAS No. 1271 19-8) in F344/N Rats (Gavage Studies). AB - Titanocene dichloride is an organometallic compound composed of two cyclopentadienyl rings, titanium, and chloride. It is used as a cocatalyst in polymerization reactions. Toxicology and carcinogenesis studies were conducted by administering titanocene dichloride (greater than 98% pure) in corn oil by gavage to groups of F344/N rats for 14 days, 13 weeks, and 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium and in Chinese hamster ovary cells. 14-Day and 13-Week Studies: In the 14-day studies, titanocene dichloride was administered at doses of 0, 65, 125, 250, 500, or 1,000 mg/kg. All high-dose rats and four of the five male and two of the five female rats given 500 mg/kg died during the studies. A dose-related decrease in body weight gain was seen in rats given 125, 250, 500, and 1,000 mg/kg. Lesions related to chemical administration included hepatocellular necrosis, tubule necrosis in the kidney, erosions and ulcers of the glandular stomach, and hyperplasia of the forestomach epithelium. The 13-week studies were conducted by administering titanocene dichloride at doses of 0, 8, 16, 31, 62, or 125 mg/kg. One female rat in the 125 mg/kg dose group died from chemical toxicity during the fourth week of the studies. Body weight gain was lower in rats given 62 or 125 mg/kg than in control groups. Treatment-associated histopathologic lesions were seen in the stomachs of high-dose males and all groups of females given titanocene dichloride. These lesions included hyperplasia and metaplasia of the glandular stomach and hyperplasia and hyperkeratosis of the forestomach. Body Weight and Survival in the 2-Year Studies: The doses selected for the 2-year studies in rats (0, 25, and 50 mg/kg) were based on the potentially life-threatening nature of the glandular stomach lesions and the decreased body weight gain compared to controls seen in the 62 and 125 mg/kg dose groups in the 13-week studies. The final mean body weights of high-dose males and females were 91% and 89% of controls, respectively. The 2-year survival rates for males in the control, low-, and high dose groups were 41/60, 30/60, and 24/60; survival rates for female rats were 37/60, 30/61, and 31/60. Nonneoplastic and Neoplastic Effects in the 2-Year Studies: The principal toxic effects associated with the administration of titanocene dichloride for 2 years occurred in the stomach. The lesions in the stomach were seen at the 15-month interim evaluations and were similar to, but less severe than, those observed at 2 years. The lesions included focal erosions of the glandular mucosa with an associated inflammatory response, hyperplasia and metaplasia of the epithelium of the fundic glands, and fibrosis of the lamina propria and submucosa. Forestomach lesions included focal acanthosis (hyperplasia) and hyperkeratosis of the stratified squamous epithelium. Squamous cell papillomas of the forestomach were seen in four low-dose males, one high dose male, one low-dose female, and two high-dose females; none were observed in controls. A squamous cell carcinoma of the forestomach occurred in one low-dose male and a benign basosquamous tumor occurred in one high-dose male. Accumulations of macrophages with blue-gray pigment believed to contain titanium were present in many organs of dosed rats including the gastrointestinal tract, liver, lung, and lymph nodes. A dose-related increase in the incidence of inflammation of the nasal mucosa and lung also occurred and was attributed to reflux and/or regurgitation and aspiration of gavage solution due to the severe stomach lesions. Genetic Toxicology: Titanocene dichloride was mutagenic in Salmonella typhimurium strain TA100 in the absence of exogenous metabolic activation (S9); it was not mutagenic in TA100 with S9, nor was it mutagenic in TA1535, TA1537, or TA98 with or without S9. Titanocene dichloride did not induce sister chromatid exchanges or chromosomal aberrations in Chinese hamster ovary cells, with or without S9. Conclusions: Under the conditions of these 2-year gavage studies, there was equivocal, there was equivocal evidence of carcinogenic activity of titanocene dichloride in male F344/N rats based on a marginal increase in the incidence of forestomach squamous cell papillomas, squamous cell carcinoma, and basosquamous tumor benign. There was equivocal evidence of carcinogenic activity of titanocene dichloride in female F344/N rats based on a marginal increase in the incidence of forestomach squamous cell papillomas. Nonneoplastic lesions associated with the administration of titanocene dichloride for up to 2 years included erosions and inflammation of the gastric mucosa, hyperplasia and metaplasia of the fundic glands with fibrosis of the lamina propria in the glandular stomach, and acanthosis (hyperplasia) and hyperkeratosis of the forestomach epithelium. Synonyms: Titanium ferrocene; biscyclopentadienyltitanium dichloride; dichlorodi-p-cyclopentadienyltitanium; dichlorobis(h5-2,4-cyclopentadien-1-yl)titanium; dicyclopentadienyltitanium dichloride; dichlorodicyclopentadienyltitanium; dichlorotitanocene; dicyclopentadienyldichlorotitanium; dichlorobis(p-cyclopentadienyl)titanium; bis (h5-cyclopentadienyl) titanium dichloride; dichlorobis(h5 cyclopentadienyl)titanium; dichlorobiscyclopentadienyl titanium; dichlorobis(1,3 cyclopentadiene)titanium; bis(cyclopentadienyl)dichlorotitanium PMID- 12637958 TI - Chronic morphine modulates the contents of the endocannabinoid, 2-arachidonoyl glycerol, in rat brain. AB - Opioids and cannabinoids are among the most widely consumed drugs of abuse in humans and the phenomena of cross-tolerance or mutual potentiation have been demonstrated between the two drugs. Several authors have suggested that both drugs share common links in their molecular mechanisms of action, although this has been a matter of controversy. Furthermore, no data exist on the possible adaptive changes in the contents of arachidonoylethanolamide (anandamide, AEA) and 2-arachidonoylglycerol (2-AG), the two major endogenous ligands for cannabinoid receptors, in morphine-tolerant rats. In the present work, we investigated the alterations in cannabinoid receptor functionality and endocannabinoid levels in rats chronically treated with morphine (5 mg/kg, s.c., twice a day for 5 days). Autoradiographic-binding studies using [(3)H]CP-55 940 revealed a slight but significant reduction in cannabinoid receptor level in the cerebellum and hippocampus of morphine-tolerant rats, while CP-55 940-stimulated [(35)S]GTPgammaS binding showed a strong decrease (40%) in receptor/G protein coupling in the limbic area of these animals. Moreover, in the same brain regions we measured, by isotope-dilution gas chromatography/mass spectrometry, the contents of AEA and 2-AG. Chronic morphine exposure produced a strong reduction in 2-AG contents without changes in AEA levels in several brain regions (ie striatum, cortex, hippocampus, limbic area, and hypothalamus). These findings clearly demonstrate that prolonged activation of opioid receptors could alter the cannabinoid system, in terms of both receptor functionality and endocannabinoid levels, and suggest the involvement of this system, alone or in combination with other mediators, in the phenomenon of morphine tolerance. PMID- 12637961 TI - NTP Toxicology and Carcinogenesis Studies of Polybrominated Biphenyls (CAS No. 67774-32-7)(Firemaster FF-1(R)) in F344/N Rats and B6C3F1 Mice (Feed Studies). AB - Polybrominated biphenyls are synthetic chemicals used as flame retardants. The technical product used in these studies, Firemaster FF-1(R)), is a mixture of brominated biphenyls. Firemaster FF-1(R)) is a known liver carcinogen in rats and mice and is one of three compounds chosen by the National Toxicology Program to investigate the potential value of perinatal exposures in assessing chemical carcinogenicity. Chronic toxicity and carcinogenicity studies of polybrominated biphenyls (Firemaster FF-1(R)) were conducted in F344/N rats and B6C3F1 mice of each sex. The studies were designed to determine: a) the effects of polybrominated biphenyls in rats and mice receiving adult ( F1) exposure only (a typical carcinogenicity study), b) the toxic and carcinogenic effects of polybrominated biphenyls in rats and mice receiving perinatal (F0) exposure only (dietary exposure of dams prior to breeding and throughout gestation and lactation), and c) the effects of combined perinatal and adult exposure to polybrominated biphenyls. STUDIES IN F344/N RATS: The exposure levels selected for F1 exposure, based on studies of polybrominated biphenyls in the literature, were 3, 10, and 30 ppm. In a preliminary study to determine the perinatal dietary concentrations for the 2-year study, female rats were administered 1 to 30 ppm polybrominated biphenyls in the feed beginning 60 days prior to breeding and continuing throughout gestation, lactation, and up to 4 weeks postweaning. The mean preweaning litter weight of the 30 ppm group was less than 80% of the mean litter weight of the control group at days 0, 4, and 12. At weaning, the mean weight of litters in this group was 80% of the control group mean. The final mean body weights (28 days after weaning) of males and females receiving 30 ppm were 13% to 19% lower than the final mean body weights of the controls. Therefore, dietary concentrations of 0, 1, 3, and 10 ppm were selected for the F0 exposure levels in the 2-year study. The eight F0 F1 exposure combinations selected for the 2-year study are shown in the following table (see page 6 of full technical report). Adult-Only Exposure The major organ affected by toxicity of polybrominated biphenyls was the liver. Rats evaluated at 9 months had decreased body weights, hepatomegaly, nonneoplastic histopathologic changes in the liver, mild anemia, increases in serum cholesterol concentrations, and decreases in serum triglyceride concentrations (males only). In rats receiving adult only exposure (F0 F1 concentrations of 0:10 or 0:30 ppm), there were no significant effects on survival. Mean body weights were significantly reduced in 0:10 and 0:30 ppm male rats and in 0:30 ppm female rats. Males and females exposed to 0:10 or 0:30 ppm had increased incidences of hepatocellular neoplasms (males: 0:0 ppm, 1/50; 0:10 ppm, 12/49; 0:30 ppm, 41/50; females: 0/50,12/50, 39/50). Increased incidences of the following nonneoplastic lesions were associated with the administration of polybrominated biphenyls: eosinophilic foci, cytoplasmic vacuolization, oval cell hyperplasia, and hypertrophy in the liver of males and females; acanthosis, inflammation, and ulceration of the forestomach in exposed males; and cystic endometrial hyperplasia of the uterus in 0:30 ppm females. Perinatal-Only Exposure For rats receiving only perinatal exposure (10:0 ppm), there were no changes in survival or body weights compared to the 0:0 ppm control groups. In female rats, there were no effects on neoplasm incidences, but perinatal exposure was associated with a marginally increased incidence of hepatocellular adenoma in male rats (0:0 ppm, 1/50; 10:0 ppm, 5/50). The incidences of nonneoplastic lesions in the liver were increased in exposed males (eosinophilic foci and cytoplasmic vacuolization) and females (eosinophilic foci). Combined Perinatal and Adult Exposure Combined perinatal and adult exposure resulted in marginally reduced survival compared to the 0:0 ppm control group for male rats in the 3:10, 10:10, and 10:30 ppm groups. No significant survival differences were obseant survival differences were observed in female rats. The final mean body weights of male and female rats receiving 3:10,10:10, or 10:30 ppm were lower than those of the 0:0 ppm controls. In male rats, there were no enhancing effects of combined perinatal and adult exposure on the incidence of hepatocellular neoplasms. However, perinatal exposure enhanced the development of liver neoplasms in female rats receiving 10 or 30 ppm adult exposure. A combined analysis of all male and female exposure groups also revealed increased incidences of mononuclear cell leukemia that were considered related to polybrominated biphenyls exposure. STUDIES IN B6C3F1 MICE: The exposure levels selected for the F1 exposure, based on studies of polybrominated biphenyls in the literature, were 3,10, and 30 ppm. In a preliminary study to determine the perinatal dietary concentrations for the 2-year study, female C57BL/6N mice were exposed to 1 to 30 ppm polybrominated biphenyls in the feed beginning 60 days before breeding to C3H/HeN males, continuing throughout gestation and lactation and up to 4 weeks postweaning. There were no clear chemical-related effects on survival or growth at any phase of the study; therefore, 0, 3,10, and 30 ppm dietary concentrations were selected for the F0 exposure levels in the 2-year study. The eight F0 F1 exposure combinations selected for the 2-year study are shown in the table below (see page 7 of full technical report). Adult-Only Exposure The major organ affected by toxicity of polybrominated biphenyls was the liver. Animals evaluated at 9 months had lower body weights than the controls, hepatomegaly, and histopathologic changes in the liver. In mice receiving adult-only exposure, no males or females in the 0:30 ppm group survived to the end of the study. Neither survival nor body weights were affected in the 0:10 ppm groups. Males and females receiving 0:10 or 0:30 ppm had markedly increased incidences of hepatocellular neoplasms (males: 0:0 ppm, 16/50; 0:10 ppm, 48/49; 0:30 ppm, 48/50; females: 5/50, 42/50, 47/48). Increased incidences of nonneoplastic liver lesions including cytomegaly (hypertrophy), fatty change (cytoplasmic vacuolization), bile duct hyperplasia, eosinophilic and clear cell foci, and necrosis of individual hepatocytes were related to treatment with polybrominated biphenyls. Increased incidences and severity of chronic nephropathy in the kidney and excessive hematopoiesis in the spleen of 0:30 ppm males and females were also considered to be related to exposure to polybrominated biphenyls. Perinatal-Only Exposure There were no survival or body weight differences in mice receiving only perinatal exposure (30:0 ppm). Perinatal exposure resulted in significantly increased incidences of hepatocellular neoplasms in males and females. The incidences of nonneoplastic lesions (cytomegaly, eosinophilic foci, clear cell foci) were increased in males and females. Combined Perinatal and Adult Exposure Combined perinatal and adult exposure resulted in markedly reduced survival for females in the 30:10 ppm group; no mice receiving 30:30 ppm survived to the end of the study. In those groups receiving adult exposure of 30 ppm, mean body weights were not affected. The incidence of hepatocellular neoplasms in male and female mice was significantly increased. At the 9-month interim evaluation the incidence of hepatocellular adenomas was significantly increased in males (0:30 ppm, 1/10; 30:30 ppm, 7/10). The incidence of hepatocellular adenomas in 30:30 ppm females was similar to that of 0:30 ppm females (0:30 ppm, 0/10; 30:30 ppm, 3/10). At the end of the study the incidence of hepatocellular adenomas in males was statistically increased (0:30 ppm, 42/50; 30:30 ppm, 48/50). The incidence of hepatocellular adenomas in 30:30 ppm females was statistically decreased compared to that of 0:30 ppm females (0:30 ppm, 46/48; 30:30 ppm, 41/47). It was not possible to assess the potential enhancing effect of combined perinatal and adult exposure on hepatocellular neoplasms because adult-only exposure resulted in such high (84% to 98%) liver neoplasm incidences. CONCLUSIONS: Adult Only Exposure Under the conditions of these 2-year, adult-only, dietary exposure studies, there was clear evidence of carcinogenic activity for polybrominated biphenyls in male and female F344/N rats and male and female B6C3F1 mice based on increased incidences of hepatocellular neoplasms. Perinatal-Only Exposure Perinatal exposure alone (through dietary administration of 10:0 ppm polybrominated biphenyls to the dams) had no effect on the incidences of neoplasms in female F344/N rats, but in male F344/N rats, perinatal exposure was associated with a marginally increased incidence of hepatocellular adenomas that may have been related to chemical administration. In male and female B6C3F1 mice, perinatal exposure to 30:0 ppm polybrominated biphenyls resulted in significantly increased incidences of hepatocellular neoplasms. The incidences of a number of nonneoplastic lesions in the liver (cytomegaly, eosinophilic focus, and clear cell focus) were increased in male and female B6C3F1 mice. Combined Perinatal and Adult Exposure Combined perinatal and adult dietary exposure to polybrominated biphenyls confirmed findings of the adult-only exposures for the increased incidences of hepatocellular neoplasms in F344/N rats and B6C3F1 mice. In male F344/N rats, there were no enhancing effects of combined perinatal and adult exposure. However, perinatal exposure enhanced the susceptibility of female F344/N rats receiving adult exposure of 10 or 30 ppm to the induction of liver neoplasms. For male and female F344/N rats, a combined analysis of the incidences of leukemia in the adult-only, perinatal-only, and combined perinatal and adult exposure groups revealed an apparent association between increasing incidences of mononuclear cell leukemia and exposure to polybrominated biphenyls. In male and female B6C3F1 mice, it was not possible to adequately assess the enhancing effects of combined perinatal and adult exposure on hepatocellular neoplasms, because adult-only exposure to 10 or 30 ppm polybrominated biphenyls resulted in high incidences (84% to 98%) of liver neoplasms. However, with increased perinatal exposure, there were increases in the numbers of B6C3F1 mice with hepatocellular carcinomas and in the numbers of B6C3F1 mice with multiple hepatocellular adenomas, which suggests an enhancement of polybrominated biphenyls-related hepatocellular carcinogenicity associated with perinatal exposure. Synonyms: PBBs; polybrominated biphenyl mixture; hexabromobiphenyl (technical grade); brominated biphenyls; polybromobiphenyls PMID- 12637962 TI - NTP Toxicology and Carcinogenesis Studies of C.I. Direct Blue 15 (CAS No. 2429-74 5) in F344 Rats (Drinking Water Studies). AB - C.I. Direct Blue 15 is one of five chemicals being evaluated in 2-year carcinogenicity and toxicity studies as part of the NTP's Benzidine Dye Initiative. This Initiative was designed to evaluate representative benzidine congeners, benzidine congener-derived dyes, and benzidine-derived dyes. The dye, industrial grade C.I. Direct Blue 15, was chosen for study as a product to which workers are potentially exposed. Because of the high salt content, the dye was desalted prior to use. The purity was determined to be approximately 50%, with high-performance liquid chromatography indicating one major peak and approximately 35 impurities. Toxicology and carcinogenesis studies were conducted by administering the dye, C.I. Direct Blue 15, in drinking water to groups of F344/N rats of each sex for 14 days, 13 weeks, or 22 months. Planned as 24-month studies, the 22-month studies were terminated early because of rapidly declining animal survival, which was due primarily to neoplasia. These studies were performed only in rats because studies of benzidine congeners were being performed in mice at the National Center for Toxicological Research (NCTR). Genetic toxicology studies were conducted in Salmonella typhimurium and Chinese hamster ovary cells. 14-Day Studies: Rats were given C.I. Direct Blue 15 in drinking water at doses of 1,250, 2,500, 5,000, 10,000, or 30,000 ppm. All control and treated rats survived. Body weight gain in high-dose females was less than that in controls. Water consumption declined as the dose increased. Male and female rats receiving 30,000 ppm had slight degeneration and necrosis of individual hepatocytes in the liver, and females also had mild to moderate renal tubule degeneration and thymic lymphoid depletion. 13-Week Studies: C.I. Direct Blue 15 was administered in drinking water at doses of 0, 1,250, 2,500, 5,000, 10,000, or 30,000 ppm to male rats, and at doses of 0, 630, 1,250, 2,500, 5,000, or 10,000 ppm to female rats. Seven of 10 male rats receiving 30,000 ppm died; all rats in the other groups survived until the end of the studies. Mean final body weights of males receiving 10,000 or 30,000 ppm were 92% and 69% of those of controls, and mean final body weights of females receiving 5,000 or 10,000 ppm were 97% and 94% of those of controls. Tissues from treated animals were stained blue. Compound-related lesions were seen in the kidney and liver of male rats given 30,000 ppm and in the kidney of males and females given 10,000 ppm. The renal lesions included necrosis, degeneration, pigmentation and regeneration of the tubule epithelium, and tubule mineralization. Liver lesions included centrilobular hepatocellular degeneration, fatty metamorphosis, and individual cell necrosis with slight periportal hepatocellular hypertrophy. Lymphoid depletion in the thymus was also seen in the high-dose males. Based on the results of the 14-day and 13-week studies, the high dose chosen for the 22-month studies was 2,500 ppm. 22-Month Studies: At study initiation, 70 rats of each sex were given 0 or 2,500 ppm C.I. Direct Blue 15, 45 rats of each sex were given 630 ppm, and 75 rats of each sex were given 1,250 ppm. Interim evaluations were made at 9 and 15 months. The average amounts of compound consumed per day by the six dose groups after week 52 of the studies were estimated to be 45, 90, and 215 mg/kg for male rats and 50, 100, and 200 mg/kg for female rats. Survival and Body Weights: The studies were terminated at 22 months due to extensive mortality associated with chemical-related neoplasia. Survival of control, 630, 1,250, and 2,500 ppm males at 22 months was 37/50, 8/35, 11/65, and 2/50; survival of females was 40/50, 13/35, 22/65, and 4/50. At 22 months, the mean final body weights of the 630, 1,250, and 2,500 ppm groups were 95%, 91%, and 81% of those of the control for male rats and 91% of those of the control for all female dose groups. Histopathologic Effects in the 22-Month Studies: At the 9-month interim evaluations, one adenoma of the Zymbal's gland was seen in a high-dose male rat, and three carcinmbal's gland was seen in a high-dose male rat, and three carcinomas of the clitoral gland were seen in the high-dose females. At the 15 month interim evaluations, Zymbal's gland neoplasms were seen in low- and high dose males and all treated female dose groups. Mid- and high-dose males and females also had preputial or clitoral gland neoplasms, and a few neoplasms were present in the skin, small and large intestine, liver, and oral cavity of treated animals at 15 months. At the end of the study, neoplasms related to chemical administration were found in the Zymbal's gland, skin, oral cavity, and the preputial or clitoral gland in both male and female rats. Neoplasms related to chemical administration were also seen at other sites including the small and large intestine, liver, uterus, and brain. The incidence of mononuclear cell leukemia was also increased in treated rats. Genetic Toxicology: C.I. Direct Blue 15 was not mutagenic in Salmonella typhimurium strains TA100, TA1535, TA1537, and TA98 when tested in a standard preincubation protocol with or without exogenous metabolic activation; however, when a specialized reductive metabolism protocol was used, C.I. Direct Blue demonstrated mutagenic activity in Salmonella strain TA1538. C.I. Direct Blue 15 did not induce sister chromatid exchanges or chromosomal aberrations in Chinese hamster ovary cells with or without S9 activation; reductive metabolism was not used in these cytogenetic tests. Conclusions: Under the conditions of these 22-month drinking water studies, there was clear evidence of carcinogenic activity of C.I. Direct Blue 15 (desalted industrial grade) in male F344/N rats, as indicated by benign and malignant neoplasms of the skin, Zymbal's gland, preputial gland, liver, oral cavity, and small and large intestine. Increased incidences of mononuclear cell leukemia and neoplasms of the brain may have been related to chemical administration. There was clear evidence of carcinogenic activity of C.I. Direct Blue 15 in female F344/N rats, as indicated by benign and malignant neoplasms of the skin, Zymbal's gland, clitoral gland, liver, oral cavity, small and large intestine, and uterus, and by mononuclear cell leukemia. Synonyms: Airedale Blue D, Aizen Direct Sky Blue 5BH, Amanil Sky Blue, Atlantic Sky Blue A, Atul Direct Sky Blue, Azine Sky Blue 5B, Belamine Sky Blue A, Benzanil Sky Blue, Benzo Sky Blue S, Benzo Sky Blue A-CF, Cartasol Blue 2GF, Chloramine Sky Blue A, Chloramine Sky Blue 4B, Chrome Leather Pure Blue, C.I. 24400, Cresotine Pure Blue, Diacotton Sky Blue 5B, Diamine Blue 6B, Diamine Sky Blue, Diaphtamine Pure Blue, Diazol Pure Blue 4B, 3,3'-[(3,3'-dimethoxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxy-2, naphthalenedisulfonic acid] tetrasodium salt, Diphenyl Brilliant Blue, Diphenyl Sky Blue 6B, Direct Blue 10G, Direct Blue HH, Direct Pure Blue, Direct Pure Blue M, Direct Sky Blue (6CI), Direct Sky Blue A, Direct Sky Blue 5B, Enianil Pure Blue AN, Fenamin Sky Blue, Hispamin Sky Blue 3B, Kayafect Blue Y, Kayaku Direct Sky Blue 5B, Mitsui Direct Sky Blue 5B, Naphtamine Blue 10G, Niagara Blue 4B, Niagara Sky Blue, Nippon Direct Sky Blue, Nitto Direct Sky Blue 5B, Paper Blue S, Phenamine Sky Blue A, Pontamine Sky Blue 5BX, Shikiso Direct Sky Blue 5B, Sky Blue 4B, Sky Blue 5B, Tertrodirect Blue F, Vondacel Blue HH PMID- 12637963 TI - NTP Toxicology and Carcinogenesis Studies of Monochloroacetic Acid (CAS No. 79-11 8) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Monochloroacetic acid, a colorless crystalline material, is used as a postemergence contact herbicide and as an intermediate in the synthesis of other organic compounds. Toxicology and carcinogenicity studies were conducted by administering monochloroacetic acid (99% pure) in deionized water by gavage to groups of F344/N rats and B6C3F1 mice of each sex once daily, 5 days per week for 16 days, 13 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, mouse lymphoma L5178Y cells, Chinese hamster ovary cells, and Drosophila melanogaster. 16-Day Studies: Groups of five rats of each sex received 0, 7.5, 15, 30, 60, or 120 mg monochloroacetic acid/kg body weight. Doses administered to mice were 0, 15, 30, 60, 120, or 240 mg/kg to groups of five males and 0, 30, 60, 120, 240, or 480 mg/kg to groups of five females. One of five male rats given 120 mg/kg died during the studies. Clear nasal discharge, lacrimation, or both, were observed in all groups of male and female rats receiving monochloroacetic acid. No compound-related gross lesions were observed in rats. All male mice given 240 mg/kg and all females given 240 or 480 mg/kg died during the studies. Hypoactivity, piloerection, ataxia, and lacrimation were observed in mice given 240 or 480 mg/kg. No compound-related gross lesions were observed in mice at necropsy. 13-Week Studies: Groups of 20 rats of each sex received 0, 30, 60, 90, 120, or 150 mg/kg monochloroacetic acid, and groups of 20 mice of each sex received doses of 0, 25, 50, 100, 150, or 200 mg/kg. Three to five animals in each dose group were killed at weeks 4 and 8 for the evaluation of hematology parameters. Compound-related deaths occurred in rats in the three highest dose groups (all males given 120 or 150 mg/kg, 9/10 males given 90 mg/kg, and all females given 90 to 150 mg/kg) and in mice given 200 mg/kg (all males and 2/10 females). Final mean body weights of surviving rats and mice receiving monochloroacetic acid were similar to those of controls. In rats, dose-related increases in blood urea nitrogen, alanine aminotransferase, and aspartate aminotransferase levels were observed, and relative liver and kidney weights were elevated. There were no compound-related changes in the various hematologic or clinical pathology parameters in mice. A dose-related increase in the incidence and severity of cardiomyopathy was observed in male and female rats receiving monochloroacetic acid, and hepatocellular cytoplasmic vacuolization was observed in the high-dose mice that died during the studies. 2-Year Studies: Based on the mortality and compound-related histopathologic lesions observed in the 13-week studies, doses selected for the 2-year studies of monochloroacetic acid were 0, 15, or 30 mg/kg, administered to groups of 70 rats of each sex, and 0, 50, or 100 mg/kg, administered to groups of 60 mice of each sex. Interim evaluations were conducted on 10 rats per dose group after 6 months of treatment with monochloroacetic acid and on seven rats per dose group after 15 months of treatment. Body Weight and Survival in the 2-Year Studies: Mean body weights of low- and high-dose female and low-dose male rats receiving monochloroacetic acid were within 10% of those of controls throughout the studies; however, after week 30, the mean body weights of high-dose male rats were 4% to 8% less than those of controls. In mice, the mean body weights of dosed males were similar to controls, but those of low- and high-dose females were 6% to 10% less than control values after week 52. Survival of high-dose male and dosed female rats and high-dose male mice was significantly lower than that of controls (male rats: control, 27/53; low-dose, 21/53; high-dose, 16/53; female rats: 37/53; 19/53; 26/53; male mice: 46/60; 39/60; 21/60; female mice: 42/60; 40/60; 44/60). Neoplasms and Nonneoplastic Lesions in the 2-Year Studies: There was no compound-related increase in the incidence of neoplasms or nonneoplastic lesions in rats given monochloroacetic acid for 2 years. The incidence of uterine stromal polypss. The incidence of uterine stromal polyps in low- and high-dose female rats was slightly higher than that in controls (2/60; 7/57; 10/60). However, the incidence in the controls was unusually low, and those in the dosed groups were well within the range for NTP historical controls (mean: 21%, range: 10% 38%). Further, because the only malignant stromal neoplasm occurred in a control animal, the polyps were not considered to be related to the administration of monochloroacetic acid. Similarly, there was no monochloroacetic acid-related increase in the incidence of neoplasms in male or female mice, and malignant lymphoma occurred with a significant negative trend in dosed female mice. Increases in the incidence of inflammation of the mucosa of the nasal passages, respiratory epithelial metaplasia of the olfactory epithelium of the nose, and focal squamous cell hyperplasia of the forestomach occurred in dosed male and female mice. Genetic Toxicology: Monochloroacetic acid was not mutagenic in Salmonella typhimurium strains TA100, TA1535, TA1537, or TA98, with or without exogenous metabolic activation (S9). It induced trifluorothymidine resistance in L5178Y cells in the absence of S9 and induced sister chromatid exchanges without S9 in Chinese hamster ovary cells. Monochloroacetic acid did not induce a significant increase in chromosomal aberrations in Chinese hamster ovary cells, with or without S9. Monochloroacetic acid administered in feed was negative for the induction of sex-linked recessive lethal mutations in germ cells of male Drosophila melanogaster; however, when it was administered by injection, the results were equivocal. Conclusions: Under the conditions of these 2-year gavage studies, there was no evidence of carcinogenic activity for monochloroacetic acid in male or female F344/N rats given 15 or 30 mg/kg. There was no evidence of carcinogenic activity for monochloroacetic acid in male or female B6C3F1 mice given 50 or 100 mg/kg. Monochloroacetic acid administration was associated with inflammatory lesions of the nasal mucosa, metaplasia of the olfactory epithelium, and squamous cell hyperplasia of the forestomach in male and female mice. Synonyms: Chloroacetic acid, a-chloroacetic acid, chloroethanoic acid PMID- 12637964 TI - NTP Toxicology and Carcinogenesis Studies of Probenecid (CAS No. 57-66-9) in F344/N Rats and B6C3F1 (Gavage Studies). AB - Probenecid is a white crystalline solid commonly used as a uricosuric agent in the treatment of gout. Because of its inhibitory effects on renal tubule transport processes, probenecid is also used as a therapeutic adjunct to enhance blood levels of penicillin and its action. Toxicology and carcinogenicity studies were conducted by administering probenecid (>99% pure) in corn oil by gavage to groups of F344/N rats and B6C3F1 mice of each sex once daily, 5 days per week in 14-day, 13-week, and 2-year studies. Genetic toxicology studies were conducted in Salmonella typhimurium and Chinese hamster ovary cells. 14-Day Studies: Doses used in the 14-day studies for both rats and mice were 0, 200, 400, 800, 1,600, or 3,200 mg/kg. Of the animals receiving 3,200 mg/kg, all rats, all female mice, and two of five male mice died during the studies. No deaths occurred among the other dose groups. There was a significant reduction in body weight gain in male and female rats receiving 1,600 mg/kg and in female rats receiving 800 mg/kg. No gross lesions were attributed to probenecid administration in rats or mice of either sex. 13-Week Studies: Doses used in the 13-week studies were 0, 50, 100, 200, 400, or 800 mg/kg for rats and 0, 100, 200, 400, 800, or 1,600 mg/kg for mice. No rats died during the 13-week studies. In mice, 5 of 10 males and 3 of 10 females receiving 1,600 mg/kg and 1 of 10 males receiving 800 mg/kg died during the study. Significant reductions in body weight gain occurred in male rats administered 800 mg/kg, male mice administered 1,600 mg/kg, and female mice administered 800 or 1,600 mg/kg. All dose groups of male rats and all groups of female rats receiving 100 mg/kg or more showed significant increases in absolute and/or relative liver weights compared to control groups. This change was also seen in mice receiving 200 mg/kg and greater, except female mice in the 400 mg/kg group. No compound-related lesions occurred in rats or mice of either sex. Based on compound-related deaths and suppression of body weight gains observed at higher doses in the 13-week studies, doses of 0, 100, and 400 mg/kg were used for the 2-year studies in rats and mice. These doses were administered once daily, 5 days a week for up to 103 weeks to groups of 50 males or 50 females of each species. Body Weight and Survival in the 2-Year Studies: The mean body weight of high-dose female rats was 10% to 20% lower than that of controls throughout the studies. Mean body weights for all other dosed rats and for all dosed mice were similar to those of controls throughout the 2-year studies. Survival of high-dose male rats and high-dose and low-dose male mice was significantly lower than that of controls. Survival rates after 2 years were: male rats--control, 37/50; 100 mg/kg, 34/50; 400 mg/kg, 22/50; female rats--24/50; 35/50; 19/50; male mice- 38/50; 23/50; 24/50; female mice--32/49; 32/49; 32/50. Neoplasms and Nonneoplastic Lesions in the 2-Year Studies: No chemical-related histopathologic toxic effects or increased incidence of tumors attributable to probenecid were observed in male or female rats receiving probenecid by corn oil gavage for up to 2 years. Mammary gland fibroadenomas and combined thyroid C-cell adenomas or carcinomas exhibited significant negative trends in female rats. These decreased tumor rates were associated with lower body weights. The incidence of adrenal medullary pheochromocytomas was significantly decreased in high-dose male rats. No compound-related increase in nonneoplastic lesions was observed in rats of either sex. No compound-related neoplastic effects were observed in male mice. In high-dose female mice, there were significant increases in the incidences of hepatocellular adenomas (3/48; 2/49; 14/49), but there was no corresponding increase in carcinomas (2/48; 2/49; 3/49). Treatment-related increased incidences of ovarian abscesses in female mice were causally related to Klebsiella species infection rather than directly related to chemical administration. Genetic Toxicology: Probenecid was not mutagenic in Salmonenot mutagenic in Salmonella typhimurium strain TA100, TA1535, TA1537, or TA98 with or without metabolic activation. In cytogenetic tests with Chinese hamster ovary cells, probenecid induced sister chromatid exchanges in the absence, but not in the presence of S9 activation. No induction of chromosomal aberrations was observed with or without S9. Conclusions: Under the conditions of these 2-year gavage studies, there was no evidence of carcinogenic activity of probenecid for male or female F344/N rats receiving 100 or 400 mg/kg in corn oil. There was no evidence of carcinogenic activity of probenecid for male B6C3F1 mice given 100 or 400 mg/kg probenecid in corn oil. There was some evidence of carcinogenic activity of probenecid for female B6C3F1 mice based on an increased incidence of hepatocellular adenomas. Synonyms: 4-[(Dipropylamino)sulfonyl]benzoic acid; p-(dipropylsulfamoyl)benzoic acid; p-(dipropylsulfamyl)benzoic acid Trade Names: Benacen; Benemid; Benemide; Benn; Probalan; Probecid; Proben; Probenid; Robenecid; Uricocid PMID- 12637965 TI - NTP Toxicology and Carcinogenesis Studies of Acetaminophen (CAS No. 103-90-2) in F344 Rats and B6C3F1 Mice (Feed Studies). AB - Acetaminophen is a widely consumed analgesic found in several nonprescription pharmaceuticals. Toxicology and carcinogenesis studies were conducted by administering acetaminophen (purity >99%) in feed to groups of F344/N rats and B6C3F1 mice of each sex for 14 days, 13 weeks, and 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium and Chinese hamster ovary cells. 14-DAY STUDIES: Rats were fed diets containing 0, 800, 1,600, 3,100, 6,200, or 12,500 ppm acetaminophen, and mice were fed diets containing 0, 250, 500, 1,000, 2,000, or 4,000 ppm acetaminophen. There were no deaths among any groups during the study; the final mean body weight of male rats that received 12,500 ppm was significantly lower than that of the controls. Final mean body weights of male and female mice and female rats that received acetaminophen were similar to those of the controls. Feed consumption by male and female rats that received 12,500 ppm acetaminophen was lower than that of the controls; feed consumption by all other exposed groups was higher than that of the controls. 13-WEEK STUDIES: Rats and mice were fed diets containing 0, 800, 1,600, 3,200, 6,200, 12,500, or 25,000 ppm acetaminophen. Two male and two female rats, and one male and one female mouse that received 25,000 ppm, and two male mice that received 12,500 ppm died from acetaminophen-related toxicity before the end of the studies. Final mean body weights of male and female rats and mice that received 12,500 or 25,000 ppm were lower than those of the controls. The patterns of feed consumption and reduced body weights that occurred among rats and mice that received diets containing 12,500 or 25,000 ppm were indicative of poor feed palatability. Acetaminophen-related lesions were observed in the liver (necrosis, chronic active inflammation, hepatocytomegaly), kidney (tubule cast, tubule necrosis, tubule regeneration), reproductive organs (atrophy of testis, ovary, and uterus), thymus and lymph nodes (lymphoid depletion) of rats that received 25,000 ppm, and of the live (chronic active inflammation, hepatocytomegaly) and testis (atrophy) of male rats receiving 12,500 ppm. Compound-related lesions in mice were found in the liver (hepatocytomegaly, focal calcification, pigmentation, necrosis) of males that received 6,200, 12,500, or 25,000 ppm and females that received 12,000 or 25,000 ppm. Dose selection for the 2-year studies was based on reduced body weights and the liver lesions observed in rats and mice at 12,500 and 25,000 ppm. 2-YEAR STUDIES: Diets containing 0, 600, 3,000, or 6,000 ppm acetaminophen were given continuously to groups of 60 rats and mice of each sex for up to 104 weeks. After 65 weeks of exposure, 10 animals from each group were evaluated for histopathology and for hematology, urinalysis, and clinical chemistry parameters. Survival and mean body weights of rats that received acetaminophen were similar to those of the controls throughout the study. The average severity of nephropathy was increased in exposed male and female rats. In males this was associated with an increased incidence of parathyroid hyperplasia (renal hyperparathyroidism). The incidence of focal renal tubule hyperplasia was also increased in exposed male rats. The incidence of mononuclear cell leukemia was increased in exposed female rats and was significantly increased in the 6,000 ppm group (9/50; 17/50; 15/50; 24/50). Survival of exposed and control mice was similar throughout the study. Mean body weights of mice that received acetaminophen were generally lower than those of the controls throughout the study. Although the incidence of thyroid follicular cell hyperplasia increased with dose among groups of exposed male and female mice, there was no increase in the incidence of follicular cell neoplasms. Renal tubule hyperplasia occurred in one low-dose and two high-dose males and a renal tubule adenoma was present in one low-dose and one high-dose male. GENETIC TOXICOLOGY: Acetaminophen was not mutagenic in Salmonella typhimurium strains TA100, TA1535, TA1537, or TA98 witr TA98 with or without S9. In cytogenetic tests with Chinese hamster ovary cells, acetaminophen induced sister chromatid exchanges and chromosomal aberrations in both the presence and absence of S9. CONCLUSIONS: Under the conditions of these 2 year feed studies, there was no evidence of carcinogenic activity of acetaminophen in male F344/N rats that received 600, 3,000, or 6,000 ppm. There was equivocal evidence of carcinogenic activity of acetaminophen in female F344/N rats based on increased incidences of mononuclear cell leukemia. There was no evidence of carcinogenic activity of acetaminophen in male and female B6C3F1 mice that received 600, 3,000, or 6,000 ppm. Nonneoplastic lesions associated with exposure to acetaminophen included increased severity of nephropathy and increased incidences of renal tubule hyperplasia and parathyroid hyperplasia in male rats, increased severity of nephropathy in female rats, and increased incidences of thyroid follicular cell hyperplasia in male and female mice. Synonyms: 4-Hydroxyacetanilide, N-Acetyl-p-aminophenol, Paracetamol PMID- 12637966 TI - NTP Toxicology and Carcinogenesis Studies of Sodium Fluoride (CAS No. 7681-49 4)in F344/N Rats and B6C3F1 Mice (Drinking Water Studies). AB - Sodium fluoride is a white, crystalline, water-soluble powder used in municipal water fluoridation systems, in various dental products, and in a variety of industrial applications. Toxicology and carcinogenesis studies were conducted with F344/N rats and B6C3F1 mice of each sex by incorporating sodium fluoride into the drinking water in studies lasting 14 days, 6 months, and 2 years. In addition, genetic toxicology studies were performed with Salmonella typhimurium, with mouse L5178Y cells, and with Chinese hamster ovary cells. 14-Day Studies: Rats and mice received sodium fluoride in drinking water at concentrations as high as 800 ppm. (Concentrations are expressed as sodium fluoride; fluoride ion is 45% of the sodium salt by weight.) In the high-dose groups, 5/5 male and 5/5 female rats and 2/5 male mice died; one female rat was given 400 ppm in the drinking water also died before the end of the studies. No gross lesions were attributed to sodium fluoride administration. 6-Month Studies: Rats received concentrations of sodium fluoride in drinking water as high as 300 ppm, and mice as high as 600 ppm. No rats died during the studies; however, among the mice, 4/9 high-dose males, 9/11 high-dose females, and 1/8 males in the 300 ppm group died before the end of the studies. Weight gains were less than those of controls for rats receiving 300 ppm and mice receiving 200 to 600 ppm. The teeth of rats and mice receiving the higher doses of sodium fluoride were chalky white and chipped or showed unusual wear patterns. Mice and male rats given the higher concentrations had microscopic focal degeneration of the enamel organ. Rats receiving 100 or 300 ppm sodium fluoride had minimal hyperplasia of the gastric mucosa of the stomach, and one high-dose rat of each sex had an ulcer. Acute nephrosis and/or lesions in the liver and myocardium were observed in mice that died early, and minimal alterations in bone growth/remodeling were observed in the long bones of mice receiving sodium fluoride at concentrations of 50 to 600 ppm. The sodium fluoride concentrations selected for the 2-year studies in both rats and mice were 0, 25, 100, and 175 ppm in the drinking water. These concentrations were selected based on the decreased weight gain of rats at 300 ppm and of mice at 200 ppm and above, on the incidence of gastric lesions in rats at 300 ppm in the 6-month studies, and on the absence of significant toxic effects at sodium fluoride concentrations as high as 100 ppm in an earlier 2-year study. Body Weights and Survival in the 2-Year Studies: Mean body weights of dosed and control groups of rats and mice were similar throughout the 2-year studies. Survival of rats and mice was not affected by sodium fluoride administration. Survival rates after 2 years were: male rats-control, 42/80; 25 ppm, 25/51; 100 ppm, 23/50; 175 ppm, 42/80; female rats-59/80; 31/50; 34/50; 54/81; male mice-58/79; 39/50; 37/51; 65/80; female mice-53/80; 38/52; 34/50; 52/80. Neoplastic and Nonneoplastic Effects in the 2-Year Studies: The teeth of rats and mice has a dose-dependent whitish discoloration, and male rats had an increased incidence of tooth deformities and attrition leading on occasion to malocclusion. The teeth of male and, to a lesser degree, female rats had areas of microscopic dentine dysplasia and degeneration of ameloblasts. Dentine dysplasia occurred in both dosed and control groups of male and female mice; the incidence of this lesion was significantly greater in high-dose than in control male mice. Osteosclerosis of long bones was increased in female rats given drinking water containing 175 ppm sodium fluoride. No other significant nonneoplastic lesions in rats or mice appeared related to sodium fluoride administration. Osteosarcomas of bone were observed in 1/50 male rats in the 100 ppm group and in 3/80 male rats in the 175 ppm group. None were seen in the control or 25 ppm dose groups. One other 175 ppm male rat had an extraskeletal osteosarcoma arising in the subcutaneous tissue. Osteosarcomas occur in historical control male rats at an incian incidence of 0.5% (range 0-6%). The historical incidence is not directly comparable with the incidences observed in this study because examination of bone was more comprehensive in the sodium fluoride studies than in previous NTP studies of other chemicals, and the diet used in previous studies was not controlled for fluoride content. In the current study, although the pairwise comparison of the incidence in the 175 ppm group versus that in the controls was not statistically significant, osteosarcomas occurred with a statistically significant dose-response trend, leading to the conclusion that a weak association may exist between the occurrence of these neoplasms and the administration of sodium fluoride. No other neoplastic lesions in rats or mice were considered possibly related to chemical administration. Genetic Toxicology: Sodium fluoride was negative for gene mutation induction in Salmonella typhimurium strains TA100, TA1535, TA1537, and TA98 with and without S9. In two laboratories, sodium fluoride was tested for induction of trifluorothymidine resistance in mouse L5178Y lymphoma cells; results were positive both with and without S9. Sodium fluoride was tested for cytogenetic effects in Chinese hamster ovary (CHO) cells in two laboratories. In the first laboratory, the sister chromatid exchange (SCE) test was negative with and without S9, and the chromosomal aberration (Abs) test was positive in the absence of S9; in the second laboratory, the SCE test was positive with and without S9, but no induction of Abs was observed. The laboratory that reported a negative result for Abs tested at doses below that shown to be positive at the other laboratory. Similarly, the positive SCE result was obtained at a higher dose and longer harvest time than used by the laboratory reporting the negative SCE response. Conclusions: Under the conditions of these 2-year dosed water studies, there was equivocal evidence of carcinogenic activity of sodium fluoride in male F344/N rats, based on the occurrence of a small number of osteosarcomas in dosed animals. "Equivocal evidence" is a category for uncertain findings defined as studies that are interpreted as showing a marginal increase of neoplasms that may be related to chemical administration. There was no evidence of carcinogenic activity in female F344/N rats receiving sodium fluoride at concentrations of 25, 100, or 175 ppm (11, 45, or 79 ppm fluoride) in drinking water for 2 years. There was no evidence of carcinogenic activity of sodium fluoride in male or female mice receiving sodium fluoride at concentrations of 25, 100, or 175 ppm in drinking water for 2 years. Dosed rats had lesions typical of fluorosis of the teeth and female rats receiving drinking water containing 175 ppm sodium fluoride had increased osteosclerosis of long bones. PMID- 12637967 TI - NTP Toxicology and Carcinogenesis Studies of Chlorinated Water (CAS Nos. 7782-50 5 and 7681-52-9) and Chloraminated Water (CAS No. 10599-90-3) (Deionized and Charcoal-Filtered) in F344/N Rats and B6C3F1 Mice (Drinking Water Studies). AB - Chlorine and chloramine are used as disinfectants in water supplies to prevent the spread of waterborne diseases. The U.S. Environmental Protection Agency and the U.S. Congress, through the Safe Drinking Water Acts and Amendments, initiated studies to determine the most effective way to disinfect water supplies and, at the same time, minimize any potential long-term health effects associated with direct chemical exposure or indirect chemical exposure through the formation of byproducts. As part of this evaluation, 2-year studies of chlorinated or chloraminated deionized charcoal-filtered drinking water were conducted in F344/N rats and B6C3F1 mice to determine the potential toxicity and carcinogenicity associated with prolonged exposure and eliminate possible confounding effects of byproducts of chlorination. Chlorinated Water Studies: Water containing 0, 70, 140, or 275 ppm chlorine (based on available atomic chlorine) was provided to groups of 70 F344/N rats or B6C3F1 mice of each sex for up to 2 years. Groups of 10 rats or mice of each sex were predesignated for evaluation at 14 or 15 weeks and 66 weeks. Survival at 2 years of rats and mice receiving chlorinated water was similar to that of the controls. Mean body weights of dosed male rats, high dose female rats, and dosed mice were slightly lower than those of their respective control groups. There was a dose-related decrease in water consumption by rats and mice. Water consumption by high-dose rats during the second year of the studies was 21% lower than controls for males and 23% lower than controls for females; water consumption by high-dose mice was 31% lower than controls for males and 26% lower than controls for females. The incidence of mononuclear cell leukemia in mid-dose, but not high-dose, female rats was significantly higher than that in controls (control, 8/50; low-dose, 7/50; mid-dose, 19/51; high-dose, 16/50). The proportion of female rats that died of leukemia before the end of the study and the mean time for observation of animals dying with leukemia were similar among all dose groups and controls. Although the marginal increase in leukemia incidence in the mid- and high-dose female rats suggested a possible association with the administration of chlorinated water, the incidence of leukemia was not clearly dose related. There was no indication of reduced latency of leukemia, and the incidence of leukemia in concurrent controls was less than the mean for historical controls; furthermore, there was no supporting evidence of an effect in male rats. Thus, the marginal increase in leukemia incidence in female rats was considered equivocal evidence of carcinogenic activity. There were no neoplasms or nonneoplastic lesions in male rats or in male or female mice that were clearly associated with the consumption of chlorinated water. Chloraminated Water Studies: Water containing 50, 100, or 200 ppm chloramine was provided to groups of 70 F344/N rats or B6C3F1 mice of each sex for up to 2 years. The same control groups were used for the chlorinated water and chloraminated water studies. Groups of 9 or 10 rats or mice of each sex were evaluated at 14 or 15 weeks and 66 weeks. Survival at 2 years of rats and mice receiving chloraminated water was similar to that of the controls. Mean body weights of high-dose rats and dosed mice were lower than those of their respective control groups. There was a dose-related decrease in water consumption by rats and mice. Water consumption during the second year of the studies by high dose rats was 34% lower than controls for males and 31% lower than controls for females; water consumption by high-dose mice was 42% lower than controls for males and 40% lower than controls for females. Mononuclear cell leukemia occurred with a marginally increased incidence in the mid- and high-dose female rats receiving chloraminated water (control, 8/50; low dose, 11/50; mid dose, 15/50; and high dose, 16/50). As in female rats receiving chlorinated water, the proportion of female rats that died of leukemia before the end of the study and t that died of leukemia before the end of the study and the mean time for observation of animals dying with leukemia were similar among all dose groups and controls. The marginal increase in leukemia incidence in females receiving chloraminated water was considered equivocal evidence of carcinogenic activity for the same reasons given for female rats receiving chlorinated water. There were no neoplasms or nonneoplastic lesions in male rats or in male or female mice that were clearly associated with the consumption of chloraminated water. Conclusions: Under the conditions of these 2-year drinking water studies, there was no evidence of carcinogenic activity of chlorinated water in male F344/N rats receiving 70, 140, or 275 ppm. There was equivocal evidence of carcinogenic activity of chlorinated water in female F344/N rats based on an increase in the incidence of mononuclear cell leukemia. There was no evidence of carcinogenic activity of chlorinated water in male or female B6C3F1 mice receiving 70, 140, or 275 ppm. Under the conditions of these 2-year drinking water studies, there was no evidence of carcinogenic activity of chloraminated water in male F344/N rats receiving 50, 100, or 200 ppm. There was equivocal evidence of carcinogenic activity of chloraminated water in female F344/N rats based on an increase in the incidence of mononuclear cell leukemia. There was no evidence of carcinogenic activity of chloraminated water in male or female B6C3F1 mice receiving 50, 100, or 200 ppm. PMID- 12637968 TI - NTP Toxicology and Carcinogenesis Studies of Tris(2-chloroethyl) Phosphate (CAS No. 115-96-8) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Tris(2-chloroethyl) phosphate (TRCP), a flame-retardant plasticizer used in plastics, polymeric foams, and synthetic fibers, was studied as part of the National Toxicology Program's class study of trisalkyl phosphate flame retardants. Toxicology and carcinogenesis studies were conducted by administering TRCP (approximately 98% pure) in corn oil by gavage to groups of F344/N rats and B6C3F1 mice of each sex for 16 days, 16 weeks, or 2 years. Genetic toxicology studies were performed in Salmonella typhimurium and Chinese hamster ovary (CHO) cells. 16-Day Studies: There were no chemical-related deaths, differences in final mean body weight, or histopathological lesions in rats receiving 22 to 350 mg/kg TRCP or in mice receiving 44 to 700 mg/kg TRCP for 12 doses over 16 days. Serum cholinesterase activity in female rats receiving 175 or 350 mg/kg TRCP was reduced slightly (80% of control levels), but enzyme activity in dosed male rats and in mice was similar to that in controls. 16-Week Studies: Rats received 22 to 350 mg/kg TRCP for 16 weeks (female) or 18 weeks (male). Several male and female rats in the 175 or 350 mg/kg dose groups died from chemical toxicity. Final mean body weights of female rats receiving 350 mg/kg were 20% greater than those of controls; final mean body weights of the remaining groups of dosed female rats and dosed male rats were similar. Chemical-related neuronal necrosis occurred in the hippocampus and thalamus of female rats and, to a lesser extent, of male rats. Serum cholinesterase activity was reduced in females receiving 175 or 350 mg/kg TRCP. There were no chemical-related deaths, differences in final mean body weight, or differences in cholinesterase activity in mice receiving 44 to 700 mg/kg TRCP for 16 weeks. Tubule epithelial cells with enlarged nuclei (cytomegaly and karyomegaly) were observed in the kidneys of high-dose (700 mg/kg) male and female mice. 2-Year Studies: The 2-year studies in rats were conducted by administering 0, 44, or 88 mg/kg TRCP to groups of 60 males and females, 5 days per week for up to 104 weeks; 9 or 10 rats of each dose group were evaluated at 66 weeks. The survival of high-dose male and female rats was reduced relative to that of controls. Final mean body weights of surviving rats were similar to those of controls. The principal chemical-related effects occurred in the kidney and brain of dosed rats. Focal hyperplasia of the renal tubule epithelium and renal tubule adenomas were markedly increased in male rats receiving 88 mg/kg TRCP and, to a lesser extent, in female rats (renal tubule hyperplasia, male rats: 0/50; 2/50; 24/50; female rats: 0/50; 3/50; 16/50; renal tubule adenoma, male rats: 1/50; 5/50; 24/50; female rats: 0/50; 2/50; 5/50). Renal tubule carcinomas occurred in one control and one high-dose male rat. Degenerative lesions consisting of gliosis, mineralization, hemorrhage, and/or hemosiderin accumulation occurred in the cerebrum and brain stem of more than 50% of female rats receiving 44 or 88 mg/kg TRCP; similar lesions were seen in only a few dosed males. Slightly increased incidences of thyroid gland follicular cell neoplasms (male rats: 5/50; 14/50; 13/50; female rats: 14/50; 16/50; 20/50) occurred in dosed males and females, but it is uncertain whether these were related to chemical administration. The 2-year studies in mice were conducted by administering 0, 175, or 350 mg/kg TRCP to groups of 60 males and females, 5 days per week for up to 104 weeks; 8 to 10 mice of each sex per dose group were evaluated at 66 weeks. There were no significant differences in survival between dosed and control groups of either sex, and final mean body weights of mice were similar among all groups. The principal chemical-related effects occurred in the kidney, in which nuclear enlargement (karyomegaly) of tubule epithelial cells was present in approximately 80% of high-dose mice. In the original diagnosis, renal tubule adenomas were seen in one control male, one high-dose male, and one low dose female. A carcinoma was also seen in one high-dose male. In a s seen in one high-dose male. In a subsequent examination of step sections of all the mouse kidneys, adenomas were found in one low-dose male and two high-dose males. The incidences of renal tubule neoplasms in the original and step sections combined were 1/50, 1/50, and 4/50 for males. Female mice receiving TRCP demonstrated a marginally increased incidence of neoplasms (primarily adenomas) of the harderian gland (3/50; 8/50; 7/50); in addition, three harderian gland neoplasms occurred in high-dose female mice evaluated after 66 weeks. Genetic Toxicology: TRCP was not mutagenic in Salmonella typhimurium strains TA100, TA1535, TA1537, or TA98 with or without exogenous metabolic activation (S9), and it tested negative for the induction of chromosomal aberrations in Chinese hamster ovary (CHO) cells. TRCP produced an equivocal response in the presence of S9 for the induction of sister chromatid exchanges (SCE) in CHO cells. Conclusions: Under the conditions of these 2-year gavage studies, there was clear evidence of carcinogenic activity for male and female F344/N rats receiving tris(2-chloroethyl)phosphate as shown by increased incidences of renal tubule adenomas. Thyroid follicular cell neoplasms and mononuclear cell leukemia in male and female rats may have been related to chemical administration. There was equivocal evidence of carcinogenic activity for male B6C3F1 mice as shown by a marginally increased incidence of renal tubule cell neoplasms. There was equivocal evidence of carcinogenic activity for female B6C3F1 mice as shown by a marginally increased incidence of harderian gland adenomas. Renal tubule cell hyperplasia in male and female rats and gliosis, hemorrhage, pigmentation (hemosiderin accumulation), and mineralization in the brains of female rats were associated with the administration of tris(2-chloroethyl) phosphate. Karyomegaly of renal tubule epithelial cells in male and female mice was also chemical related. Synonyms: 2 chloroethanol phosphate (3:1); tris(b-chloroethyl) phosphate Trade Names: Fyrol CEF; Disflamoll TCA; NIAX flame retardant PMID- 12637969 TI - NTP Toxicology and Carcinogenesis Studies of 3,3'-Dimethylbenzidine Dihydrochloride (CAS No. 612-82-8) in F344/N Rats (Drinking Water Studies). AB - 3,3'-Dimethylbenzidine dihydrochloride is one of five chemicals being evaluated in 2-year carcinogenicity and toxicity studies as part of the NTP's Benzidine Dye Initiative. This Initiative was designed to evaluate representative benzidine congeners, benzidine congener-derived dyes, and benzidine-derived dyes. 3,3' Dimethylbenzidine dihydrochloride was nominated for study because of the potential for human exposure during production of bisazobiphenyl dyes and because benzidine, a structurally related chemical, is a known human carcinogen. Toxicology and carcinogenesis studies were conducted by administering 3,3' dimethylbenzidine dihydrochloride (approximately 99% pure) in drinking water to groups of F344/N rats of each sex for 14 days, 13 weeks, or 9 or 14 months. The 14-month exposures were planned as 24-month exposures but were terminated early because of rapidly declining animal survival, due primarily to neoplasia. These studies were performed only in rats because similar studies were being performed in mice at the National Center for Toxicological Research (NCTR). Hematologic and serum chemical analyses and thyroid hormone determinations were conducted in conjunction with the 13-week and 9-month studies. Genetic toxicology studies were conducted in Salmonella typhimurium, Chinese hamster ovary (CHO) cells, and Drosophila melanogaster. 14-Day Studies: Rats were exposed to 3,3' dimethylbenzidine dihydrochloride in drinking water at doses ranging from 600 to 7,500 ppm. All five males and one female in the 7,500 ppm group and 1/5 males in the 5,000 ppm group died. Final mean body weights were decreased in males receiving 1,250 ppm or more and in all exposed females, and final mean body weights of animals receiving 2,500 ppm or more were lower than initial weights. Water consumption decreased with increasing chemical concentration. Compound related effects observed in rats receiving 5,000 ppm or more included minimal to slight hepatocellular necrosis, accumulation of brown pigment (presumably bile) in individual hepatocytes, increased severity of nephropathy relative to controls, and severe lymphocytic atrophy of the thymus. Treated animals also showed an increased severity of atrophy of the bone marrow relative to controls, varying degrees of lymphocytic atrophy of the mandibular and mesenteric lymph nodes and spleen, increased vacuolization and necrosis of cells of the adrenal cortex, focal acinar cell degeneration in the pancreas, and, in males, increased immature sperm forms in the testis and epididymis. 13-Week Studies: 3,3' Dimethylbenzidine dihydrochloride was administered in drinking water at doses of 300, 500, 1,000, 2,000, and 4,000 ppm. All rats receiving 4,000 ppm and 4/10 males and 1/10 females receiving 2,000 ppm died before the end of the studies. Depressions in final mean body weight relative to controls ranged from 12% to 48% for males and from 9% to 42% for females. Water consumption decreased with increasing dose. At compound concentrations of 300 to 2,000 ppm, mean water consumption was 29% to 83% of control values. Compound-related effects included an increase in the severity of nephropathy relative to controls; hepatocellular necrosis and accumulation of brown pigment (presumably bile) in sinusoidal lining cells; lymphocytic atrophy of the thymus, spleen, and mandibular and mesenteric lymph nodes; atrophy of the bone marrow in the higher-dose groups; degeneration of pancreatic acinar cells; and, in males, immature sperm forms in the testis and epididymis. Decreases in serum triiodothyronine (T3) values were observed in exposed females, and decreases in mean thyroxin (T4) concentrations in exposed males and females; no significant changes were observed in thyroid stimulating hormone (TSH) levels in exposed rats. Based on the decreased survival, reductions in water consumption and body weight gain, and chemical-induced hepatocellular and renal lesions observed in the 13-week studies, the doses selected for the 9- and 14-month drinking water studies of 3,3'-dimethylbenzidine dihydrochloride were 0, 3 3,3'-dimethylbenzidine dihydrochloride were 0, 30, 70, and 150 ppm. Seventy rats of each sex were used in the control group, 45 in the low-dose group, 75 in the mid-dose group, and 70 in the high-dose group. 9-Month Studies: Ten rats of each sex in the control and 150 ppm dose groups were evaluated after 9 months. Chemical-related effects observed in exposed animals included alveolar/bronchiolar carcinoma in one male, basal cell carcinoma of the skin in one male, a squamous cell carcinoma of the oral cavity in one female, preputial gland carcinoma in two males, clitoral gland carcinoma in three females, adenocarcinoma of the small intestine in two males, Zymbal's gland carcinoma in two males and three females, hepatocellular carcinoma in two males, and adenomatous polyps of the large intestine in three males. Other effects seen in dosed rats included focal cellular alteration in the liver, lymphoid atrophy in the spleen, and increased severity of nephropathy relative to controls. An increase in serum T3 values was observed in exposed males, and a decrease in mean T4 concentrations in exposed males and females. TSH concentrations were increased in exposed male and female rats. Body Weights and Survival in the 14-Month Studies: The average amount of 3,3'-dimethylbenzidine dihydrochloride consumed per day was approximately 1.8, 4.0, or 11.2, mg/kg for low-, mid-, or high-dose male rats and 3.0, 6.9, or 12.9 mg/kg for low-, mid-, or high-dose female rats. The mean body weight of high-dose males was about 85% of the control value by week 28. By the end of the study, mean body weights of low-, mid-, and high dose males were 97%, 92%, and 70% of the control values, respectively. Mean body weights of high- and mid-dose females were about 85% of the control values at week 32 and week 44, respectively. At the end of the study, mean body weights of exposed females were about 94%, 81%, and 74% of the control values for low-, mid-, and high-dose groups, respectively. Because of extensive neoplasia, many exposed males and females were dying or were sacrificed moribund in the first year, and all high dose males died by week 55. The studies were terminated at weeks 60 to 61, at which time the group survivals were male: control, 60/60, low dose, 41/45; mid dose, 50/75; high dose, 0/60; female: 59/60; 39/45; 32/75; 10/60. Nonneoplastic Effects in the 14-Month Studies: Increases in nonneoplastic lesions in dosed rats included cystic degeneration and foci of cellular alteration in the liver; exacerbation of nephropathy; and focal or multifocal hyperplasia of the Zymbal's gland, preputial and clitoral glands, and alveolar epithelium. Neoplastic Effects in the 14-Month Studies: Neoplasms were observed in exposed rats at many sites: skin, Zymbal's gland, preputial and clitoral glands, liver, oral cavity, small and large intestine, mammary gland, lung, brain, and mesothelium. The incidence of these neoplastic effects in male and female rats is summarized in the table at the end of this section (see page 8 of the Technical Report). Genetic Toxicology: 3,3'-Dimethylbenzidine dihydrochloride was mutagenic in Salmonella typhimurium strain TA98 with exogenous metabolic activation; it was not mutagenic in strains TA100, TA1535, or TA97 with or without activation. 3,3'-Dimethylbenzidine dihydrochloride induced sister-chromatid exchanges (CHO) and chromosomal aberrations in CHO cells in the absence of exogenous metabolic activation; these effects were not evident in test with S9 activation. Sex-linked recessive lethal mutations were induced in germ cells of adult male Drosophila melanogaster administered 3,3'-dimethylbenzidine dihydrochloride in feed or by injection. No reciprocal translocations occurred in D. melanogaster germ cells following exposure to 3,3'-dimethylbenzidine dihydrochloride. Conclusions: Under the conditions of these 14-month drinking water studies, there was clear evidence of carcinogenic activity of 3,3'-dimethylbenzidine dihydrochloride for male F344/N rats, as indicated by benign and malignant neoplasms of the skin, Zymbal's gland, preputial gland, liver, oral cavity, small and large intestine, lung, and mesothelium. Increased incidences of neoplasms of the brain may have been related to chemical administration. There was clear evidence of carcinogenic activity for female F344/N rats, as indicated by benign and malignant neoplasms of the skin, Zymbal's gland, clitoral gland, liver, oral cavity, small and large intestine, mammary gland, and lung. Increased incidences of neoplasms of the brain and mononuclear cell leukemia may have been related to chemical administration. Synonyms: o-tolidine dihydrochloride; 3,3'-dimethylbiphenyl-4,4'-diamine dihydrochloride; 3,3'-dimethylbiphenyl-4,4'-biphenyldiamine dihydrochloride; 4,4' diamino-3,3'-dimethylbiphenyl dihydrochloride PMID- 12637970 TI - NTP Toxicology and Carcinogeneis Studies of Sodium Azide (CAS: 26628-22-8) in F344 Rats (Gavage Studies). AB - Sodium azide is a white crystalline solid used in the manufacture of the explosive lead azide. It is the principal chemical used to generate nitrogen gas in automobile safety airbags and airplane escape chutes and is a broad-spectrum biocide used in both research and agriculture. Toxicology and carcinogenicity studies were conducted by administering sodium azide (greater than 99% pure) in distilled water by gavage to groups of male and female F344/N rats once daily, 5 days per week for 14 days, 13 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium and Chinese hamster ovary cells. 14-Day Studies: Rats received 0, 5, 10, 20, 40, or 80 mg/kg sodium azide. All male and female rats receiving 40 or 80 mg/kg and two of five female rats receiving 20 mg/kg died during the first week of the studies. Clinical findings of toxicity included lethargy and inactivity. No grossly observable lesions were present in any of the dose groups. 13-Week Studies: Rats received 0, 1.25, 2.5, 5, 10, or 20 mg/kg sodium azide. Seven of 9 males and all 10 females receiving 20 mg/kg died before the end of the studies. Final mean body weights of treated rats were within 10% of those of the controls. Compound-related clinical findings of toxicity in the 20 mg/kg dose groups included lethargy and labored breathing. Histopathologic lesions induced by sodium azide were limited to the brain (necrosis of the cerebrum and thalamus) and lung (congestion, hemorrhage, and edema), and were observed in rats receiving 20 mg/kg that died during the studies. Body Weights, Feed Consumption, and Survival in the 2-Year Studies: Because compound-related deaths were observed in the groups receiving 20 mg/kg in the 13-week studies, lower dose levels were used in the 2-year studies. Two-year studies were conducted by administering 0, 5, or 10 mg/kg sodium azide to groups of 60 male and 60 female rats. Dose-related depression in mean body weight was observed throughout the study period. Mean feed consumption values in low- and high-dose groups were lower than control values. Survival of high-dose rats of each sex was significantly (P<0.05) lower than controls (males-control, 24/60; low-dose, 27/60; high-dose, 9/60; females-37/60; 43/60; 21/59). The reduced survival was attributed to brain necrosis and cardiovascular collapse induced by sodium azide. Neoplastic and Nonneoplastic Effects in the 2-Year Studies: There were no compound-related increases in incidences of neoplasms in rats. Significantly decreased incidences were observed for certain neoplasms, including mononuclear cell leukemia in male rats (control, 33/60; low-dose, 28/60; high dose, 14/60), adrenal gland pheochromocytoma in male rats (26/55; 16/56; 6/54), mammary gland fibroadenoma in female rats (20/60; 11/60; 8/59), and pituitary gland neoplasms in female rats (37/60; 28/60; 17/59). These decreases reflected to some extent, but could not be attributed solely to, the reduced survival of the high-dose groups. Compound-related nonneoplastic brain lesions (necrosis of the cerebrum and thalamus) were observed at significantly (P<0.001) increased incidences in high-dose male and female rats. The increased incidence of lung congestion observed in this dose group was considered due to cardiovascular collapse secondary to brain necrosis. Genetic Toxicology: Sodium azide was mutagenic in Salmonella typhimurium strains TA100 and TA1535, with or without exogenous metabolic activation (S9); it was not mutagenic in strain TA1537 or TA98. In cytogenetic tests with Chinese hamster ovary cells, sodium azide induced sister chromatid exchanges, but not chromosomal aberrations, in the presence and the absence of S9. Conclusions: Under the conditions of these 2-year gavage studies, there was no evidence of carcinogenic activity of sodium azide in male or female F344/N rats administered 5 or 10 mg/kg. Sodium azide induced necrosis in the cerebrum and the thalamus of the brain in both male and female rats. Synonyms: Azide, Azium, Smite PMID- 12637971 TI - NTP Toxicology and Carcinogenesis Studies of Ethylene Thiourea (CAS: 96-45-7) in F344 Rats and B6C3F1 Mice (Feed Studies). AB - Ethylene thiourea is a white crystalline solid used extensively in the rubber industry as an accelerator in the vulcanization of elastomers. It is also a trace contaminant and metabolic degradation product of a widely used class of ethylene bisdithiocarbamate fungicides. Ethylene thiourea is known to produce thyroid neoplasms in rats and liver neoplasms in mice following long-term administration; thus, it was chosen by the National Toxicology Program in an investigation of the potential value of perinatal exposures in assessing chemical carcinogenicity. Chronic toxicity and carcinogenicity studies of ethylene thiourea, 99% pure, were conducted in F344/N rats and B6C3F1 mice of each sex. The studies were designed to determine 1) the effects of ethylene thiourea in rats and mice receiving adult exposure only (a typical carcinogenicity study), 2) the toxic and carcinogenic effects of ethylene thiourea on rats and mice receiving perinatal exposure only (dietary exposure of dams prior to breeding and throughout gestation and lactation), and 3) the effects of combined perinatal and adult exposure to ethylene thiourea. STUDIES IN F344/N RATS: In a preliminary study to determine the perinatal dietary concentrations for the 2-year studies, female F344/N rats were fed 0, 8, 25, 83, or 250 ppm ethylene thiourea in the feed beginning 2 weeks prior to breeding and continuing throughout gestation and lactation, and the pups were fed at these same concentrations up to 9 weeks postweaning. Based on decreased survival of rat pups between postnatal days 0 to 4 and reduction in body weight gains in male weanling rats receiving 250 ppm, dietary concentrations of 0, 9, 30, and 90 ppm were selected for the perinatal (F0) exposure levels in the 2-year studies. Groups of 10 male and 10 female rats, 8 to 9 weeks of age, were fed diets containing 0, 60, 125, 250, 500, or 750 ppm ethylene thiourea for 13 weeks to determine the adult dietary concentrations. Because of reduced weight gains and decreased feed consumption in rats receiving 500 or 750 ppm ethylene thiourea, dietary concentrations of 0, 25, 83, and 250 ppm were selected for the adult (F1) exposure during the 2-year studies. In the 2-year studies, perinatal and adult exposures to ethylene thiourea were applied separately and together to groups of male or female rats as shown in the following table. The principal toxic effects of ethylene thiourea involved the thyroid gland. Serum levels of thyroxine (T4) and/or triiodothyronine (T3) were significantly decreased in rats receiving adult concentrations of 83 or 250 ppm, and thyrotropin (thyroid stimulating hormone, TSH) was significantly increased at these concentrations. In male and female rats receiving adult-only exposure of 83 or 250 ppm, the incidences of follicular cell hyperplasia or follicular cell adenoma of the thyroid gland were significantly increased relative to the controls. The incidences of follicular cell carcinoma were significantly increased in the 250 ppm groups, and carcinomas occurred more frequently in males than in females. Exposure Groups of Rats in the 2-Year Feed Studies of Ethylene Thiourea a F1 Concentration (ppm)b F0(ppm)c 0 25 83 250 0 60 -- 60 60 9 -- 60 -- -- 30 -- -- 60 -- 90 60 -- 60 60 a Ten rats from each group were sacrificed and evaluated at 9 months b Concentration of ethylene thiourea in feed given to rats beginning at 8 weeks of age for 24 months c Concentration of ethylene thiourea in feed through breeding, gestation, and lactation until pups were 8 weeks of age Perinatal-only exposure to 90 ppm had no effect on the incidence of thyroid neoplasms in these studies, although there was a marginal increase in follicular cell hyperplasia relative to the controls. However, for groups of rats receiving combined perinatal and adult exposure (F0:F1), males and females receiving concentrations of 90:250 ppm ethylene thiourea had significantly increased incidences of thyroid follicular cell neoplasms relative to those receiving adult-only exposure to 250 ppm. Further, groups of male rats receiving 90:83 ppm 83 ppm showed a significantly increased incidence of follicular cell hyperplasia. Final mean body weights of males and survival of males and females receiving combined perinatal (90 ppm) and adult (250 ppm) exposure were lower than those receiving adult-only exposure of 250 ppm. Thus, in rats, combined perinatal and adult exposure slightly enhanced the toxicity and proliferative effects on the thyroid gland observed with adult-only exposure to ethylene thiourea. Neoplasms of the Zymbal's gland were marginally increased in rats receiving 90:250 ppm (males - 0:0, 1/50; 90:250, 5/50; females - 0:0, 1/50; 90:250, 4/50). Mononuclear cell leukemia occurred with a significant trend in groups of male and female rats receiving perinatal exposure of 90 ppm and increasing adult concentrations (90:0, 90:83, and 90:250 ppm), and for female rats without perinatal exposure (0:0, 0:83, and 0:250 ppm). The incidences of mononuclear cell leukemia in males receiving 90:83 ppm and males and females receiving 90:250 ppm were statistically significant relative to the respective 0:0 ppm groups. Low incidences of renal tubule cell adenomas occurred in most dose groups of male rats, but not in the highest dose group or the controls. STUDIES IN B6C3F1 MICE: In a preliminary study to determine the perinatal dietary concentrations for the 2-year studies, adult female C57BL/6N mice were fed 0, 33, 100, 330, or 1,000 ppm ethylene thiourea in the feed beginning 2 weeks prior to breeding and continuing throughout gestation and lactation and up to 9 weeks postweaning. Because of reduced survival of mouse pups at postnatal day 28 and lower final mean body weights in weanlings receiving perinatal exposure of 1,000 ppm, dietary concentrations of 0, 33, 110, and 330 ppm were selected for the perinatal exposure levels in the 2-year studies. Groups of 10 male and 10 female mice, 8 to 9 weeks of age, were fed diets containing 0, 125, 250, 500, 1,000, or 2,000 ppm ethylene thiourea for 13 weeks to determine the adult dietary concentrations. Moderately severe diffuse follicular cell hyperplasia in the thyroid gland and centrilobular cytomegaly of the liver occurred in mice receiving 2,000 ppm. Because the severity of the thyroid lesion (and degree of hypothyroidism) at this concentration was considered potentially life threatening in 2-year studies, dietary concentrations of 0, 100, 330, and 1,000 ppm ethylene thiourea were selected for adult exposure during the 2-year studies. In the 2-year studies, perinatal and adult exposures to ethylene thiourea were applied separately and together to groups of male or female mice as shown in the following table. Exposure Groups of Mice in the 2-Year Feed Studies of Ethylene Thiourea a F1 Concentration (ppm)b F0(ppm)c 0 100 330 1,000 0 69 -- 60 60 33 -- 34/29d -- -- 110 -- -- 60 -- 330 60 -- 60 60 a Ten mice from each group except the 33:100 ppm group were sacrificed and evaluated at 9 months b Concentration of ethylene thiourea in feed given to mice beginning at 8 weeks of age for 24 months c Concentration of ethylene thiourea in feed through breeding, gestation, and lactation until pups were 8 weeks of age d 34 males and 29 females assigned to group The principal toxic effects of ethylene thiourea in mice occurred in the thyroid gland, liver, and pituitary gland. Serum levels of T3 were significantly decreased in groups of mice receiving adult concentrations of 1,000 ppm; TSH was significantly increased in mice receiving 330 and 1,000 ppm. The incidences of follicular cell hyperplasia and neoplasia increased principally in males receiving 1,000 ppm and in females receiving 330 or 1,000 ppm. Follicular cell carcinomas were significantly increased in mice receiving 1,000 ppm. Incidences of centrilobular hepatocellular cytomegaly (males and females), hepatocellular adenoma (females), hepatocellular carcinoma (males and females), and adenoma or carcinoma combined (males and females) also were significantly increased in mice receiving F1 concentrations of 330 or 1,000 ppm. In the pituitary gland, incidences of focal hyperplasia (males) or adenoma (males and females) of the pars distalis were significantly increased in groups of mice receiving 1,000 ppm ethylene thiourea. Perinatal exposure to concentrations of 330 ppm had no effect on the incidences of nonneoplastic lesions or neoplasms in mice. For groups of mice receiving combined perinatal and adult exposure, females receiving F0:F1 concentrations of 330:330 ppm had significantly increased incidences of follicular cell adenoma relative to those receiving adult-only exposure to 330 ppm. Similarly, male mice receiving F0:F1 concentrations of 330:330 ppm had significantly increased incidences of follicular cell hyperplasia. Thus, in mice, perinatal exposure slightly enhanced the proliferative effects on the thyroid gland of adult exposure. There were no effects of perinatal exposure in mice at sites other than in the thyroid gland. CONCLUSIONS: 2-Year Adult-Only Exposure:Under the conditions of these 2-year adult-only dietary exposures, there was clear evidence of carcinogenic activity of ethylene thiourea in male and female F344/N rats, as shown by increased incidences of thyroid follicular cell neoplasms. There was clear evidence of carcinogenic activity of ethylene thiourea in male and female B6C3F1 mice as shown by increased incidences of thyroid follicular cell neoplasms, hepatocellular neoplasms, and adenomas of the pars distalis of the pituitary gland. Nonneoplastic lesions associated with the administration of ethylene thiourea included follicular cell hyperplasia in rats and mice and follicular cell cytoplasmic vacuolation, centrilobular hepatocellular cytomegaly, and focal hyperplasia of the pars distalis of the pituitary gland in mice. Other effects associated with the administration of ethylene thiourea included decreased serum levels of T4 and/or T3 in rats and increased serum levels of TSH in rats and mice. Perinatal-Only Exposure:Perinatal exposure alone had no effect on the incidences of neoplasms in rats or mice after 2 years. Animals may have been able to tolerate higher perinatal exposure concentrations. Combined Perinatal and 2 Year Adult Exposures:Combined perinatal and 2-year adult dietary exposure to ethylene thiourea confirmed the findings of the 2-year adult-only exposures for the incidences of neoplasms in the thyroid gland of rats and mice and the liver and pituitary gland of mice. In male and female rats, combined perinatal and adult exposure to 90:250 ppm was associated with marginal increases, relative to the untreated (0:0 ppm) controls, in Zymbal's gland neoplasms and mononuclear cell leukemia, which may have been related to chemical administration. In rats receiving adult exposure to 250 ppm ethylene thiourea, perinatal exposure to 90 ppm was associated with a slightly enhanced incidence of thyroid neoplasms compared to adult-only exposure. However, increasing perinatal exposure from 0 to 90 ppm had no effect on incidences of thyroid neoplasms in rats receiving adult exposure to 83 ppm. Increasing perinatal exposure from 0 to 330 ppm was associated with a marginally increased incidence of thyroid neoplasms in female mice receiving adult exposure to 330 ppm, but there were no enhancing effects of perinatal exposure in mice receiving adult exposure to 1,000 ppm. Synonyms: 2 Imidazolidinethione; Imidazoline-2-thiol; 2-mercaptoimidazoline; N,N' ethylenethiourea; 1,3-ethylenethiourea; 2-imadazoline-2-thiol PMID- 12637972 TI - NTP Toxicology and Carcinogenesis Studies of dl-Amphetamine Sulfate (CAS No. 60 13-9) in F344/N Rats and B6C3F1 Mice (Feed Studies). AB - dl-Amphetamine sulfate is used for the treatment of narcolepsy in adults and behavioral syndromes in children. Toxicology and carcinogenesis studies were conducted by administering dl-amphetamine sulfate (USP grade) in feed to groups of F344/N rats and B6C3F1 mice of each sex for 14 days, 13 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium and Chinese hamster ovary (CHO) cells. 14-Day Studies: The chemical was administered at dietary concentrations of 0, 47, 94, 188, 375, or 750 ppm for rats and 0, 125, 250, 500, 1,000, or 2,000 ppm for mice. Decreased body weight gain was seen at the higher concentrations, but no chemical-related deaths or toxic lesions were observed. 13-Week Studies: The chemical was administered at dietary concentrations of 0, 47, 94, 188, 375, or 750 ppm for rats and 0, 125, 250, 500, 1,000, or 2,000 ppm for mice. None of the rats died, but 6/10 male mice and 7/10 female mice that received 2,000 ppm, 3/10 male mice that received 1,000 ppm, and 8/10 male mice that received 500 ppm died before the end of the studies. Decreased body weight gain and hyperactivity were seen in dosed rats and mice. Final body weights of rats receiving 188 ppm or more were 62% to 89% those of controls, and final body weights of mice receiving 250 ppm or more were 70% to 86% those of controls. There were no lesions that were considered to be a primary effect of the chemical. Based on decreased body weight gain and hyperactivity in the 13-week studies, 2-year studies were conducted by feeding diets containing 0, 20 or 100 ppm dl-amphetamine sulfate to groups of 50 rats or 50 mice of each sex. Body Weights and Survival in the 2-Year Studies: No significant differences in survival were observed between any groups of rats or mice (male rats: control, 30/50; low dose, 31/50; high dose, 33/50; female rats: 33/50; 42/50; 37/50; male mice: 48/50; 48/50; 49/50; female mice: 35/50; 36/50; 44/50). Final body weights of dosed rats and mice were decreased relative to those of controls. Final body weights were 92% and 86% those of controls for low- and high-dose male rats, 89% and 70% those of controls for low- and high-dose female rats, 85% and 72% those of controls for low- and high-dose male mice, and 81% and 66% those of controls for low- and high-dose female mice. Hyperactivity was observed in all dosed groups. Feed consumption was similar among control and exposed groups with the exception of high-dose female rats (84% of controls) and high-dose male mice, for which hyperactivity resulted in scattering of feed and overestimation of feed consumption. The average amount of dl-amphetamine sulfate consumed per day was estimated to be 1 or 5 mg/kg for low- and high-dose rats, 4 or 30 mg/kg for low- or high-dose male mice, and 3 or 19 mg/kg for low- or high-dose female mice. Nonneoplastic and Neoplastic Effects in the 2-Year Studies: Myelofibrosis, cataracts, and retinal atrophy in female rats, and ovarian atrophy in female mice occurred in a larger proportion of high-dose animals than in controls. Dose related increases in neoplasms did not occur in rats or mice receiving amphetamine. The administration of dl-amphetamine sulfate was associated with decreases in the incidence of total neoplasms and in the incidences of certain site-specific neoplasms, including pheochromocytomas of the adrenal gland in male rats (23/49, 15/44, 7/50), fibroadenomas of the mammary gland in female rats (21/50, 11/50, 2/50), adenomas of the anterior pituitary gland in male and female rats and female mice (male rats: 15/49, 15/48, 9/49; female rats: 31/50, 24/48, 19/50; female mice: 12/49, 6/49, 1/46), endometrial stromal polyps of the uterus of female rats (10/50, 6/50, 3/50), adenomas or carcinomas (combined) of the liver in male and female mice (male: 14/50, 12/50, 2/50; female: 5/50, 1/50, 1/47), adenomas of the harderian gland in male and female mice (male: 4/50, 2/50, 0/50; female: 5/50, 2/50, 0/47), and adenomas or carcinomas (combined) of the lung in male and female mice (male: 8/50, 3/50, 4/50; female: 8/50, 6/50 (combined) of the lung in male and female mice (male: 8/50, 3/50, 4/50; female: 8/50, 6/50, 1/47). Genetic Toxicology: dl-Amphetamine sulfate was tested for induction of gene mutations in Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537 with and without exogenous metabolic activation (S9); the only response observed was in strain TA98 in the presence of S9, and it was judged to be equivocal. No induction of sister chromatid exchanges or chromosomal aberrations occurred in Chinese hamster ovary cells treated with amphetamine sulfate in either the presence or the absence of S9. Conclusions: Under the conditions of these 2-year feed studies, there was no evidence of carcinogenic activity of dl-amphetamine sulfate for male or female F344/N rats or male or female B6C3F1 mice fed 20 or 100 ppm. The administration of dl-amphetamine sulfate was associated with decreased body weight. There were decreased incidences of total neoplasms in dosed rats and mice, of adrenal pheochromocytomas in male rats, of mammary gland fibroadenomas and uterine polyps in female rats, of pituitary gland adenomas in male and female rats and female mice, and of harderian gland adenomas, liver neoplasms, and lung neoplasms in male and female mice. Synonyms: (±)-amphetamine sulfate; (±)-2 amino-1-phenylpropane sulfate; amphetamine sulfate; deoxynorephedrine; desoxynorephedrine; (±)-a-methylphenethylamine sulfate; (±) phenisopropylamine sulfate; b-phenyl isopropylamine Trade Names: Acedron; Adipan; Adiparthrol; Aketdrin; Aktedrin; Alentol; Amfetamina; Amphaetamin; Amphamed; Amphatamin; Amphate; Amphedrine; Amphetaminum; Amphezamin; Amphoids-S; Anara; Anfetamina; Anorexine; Astedin; Benzafinyl; Benzamphetamine; Benzebar; Benzedrina; Benzedryna; Benzolone; Benzpropamine;Betafen; Betaphen; Bluzedrin; Centramina; Didrex; Dietamine; Durophet; Elastonin; Elastonon; Euphobine; Euphodine; Euphodyn; Fabedrine; Fenamin; Fenara; Fenedrin; Fenopromin; Halloo Wach; Ibiozedrine; Isamin; Isoamin; Isoamyne; Isomyn; Leodrin; Levonor; Linampheta; Mecodrin; Mimetina; Monetamine; Noclon; Norephedrane; Norphedrane; Novydrine; Oktedrin; Oraldrina; Ortenal; Orthedrin; Percomon; Pharmamedrine; Pharmedrine; Phenamine; Phenedrine; Phenopromin; Phenpromin; Profamina; Profetamine; Propenyl; Propisamine; Psychedrine; Psychedryna; Psychedrinum; Psychoton; Racephen; Rhinalator; Sedolin; Simpamina; Simpamine; Simpatedrin; Stimulan; Sympametin; Sympamine; Sympatedrine; Synsatedrine; Theptine; Vapedrine; Weckamine; Zedrine Slang for Amphetamines: bennies; benzies; cartwheels; hearts; peaches; roses PMID- 12637973 TI - NTP Toxicology and Carcinogenesis Studies of Tetranitromethane (CAS No. 509-14-8) in F344/N Rats and B6C3F1 Mice (Inhalation Studies). AB - Tetranitromethane is a volatile contaminant formed during the manufacture of TNT and has been used as a rocket fuel and biochemical reagent. Toxicology and carcinogenesis studies were conducted in F344/N rats and B6C3F1 mice of each sex by whole body exposure to tetranitromethane vapor (greater than 99% pure), 6 hours per day, 5 days per week for 14 days, 13 weeks, or 2 years. Additional groups of male mice were exposed to tetranitromethane for evaluation at 1 year. Genetic toxicology studies were performed in Salmonella typhimurium and Chinese hamster ovary (CHO) cells. Fourteen-Day Studies: Exposure concentrations ranged from 2 to 25 ppm for rats and from 2 to 50 ppm for mice. All rats exposed to 25 ppm and all mice exposed at the top concentration of 50 ppm died by day 2; reduced survival was seen in mice exposed to 25 ppm and in rats exposed to 10 ppm. Pulmonary edema in rats and inflammation of the lung in mice were seen in those animals in the 25- and 50-ppm exposure groups examined microscopically. Thirteen-Week Studies: Exposure concentrations ranged from 0.2 to 10 ppm for rats and mice. No exposure-related deaths occurred in rats. The final mean body weight of rats exposed to 10 ppm was 16% lower than that of controls for males and 6% lower for females. Exposure-related histologic effects included squamous metaplasia of the respiratory epithelium of the nasal mucosa and chronic inflammation of the lung. No deaths of mice could be clearly related to exposure to tetranitromethane. The final mean body weights of mice exposed to 5 or 10 ppm were 5% or 12% lower than that of controls for males and 9% or 12% lower for females. Exposure-related histologic effects in mice included inflammation and squamous metaplasia of the respiratory epithelium of the nasal mucosa and hyperplasia of the bronchiolar epithelium. Based on the incidences and severity of lesions in the respiratory at the higher concentrations used in the 13-week studies, exposure concentrations chosen for the 2-year studies were 0, 2, and 5 ppm for groups of 50 rats of each sex and 0, 0.5, and 2 ppm for groups of 50 mice of each sex. Additional groups of 6 or 10 male mice were exposed at concentrations of 0, 0.5, or 2 ppm for 1 year. Body Weights and Survival in the Two-Year Studies: Mean body weights of male and female rats exposed to 5 ppm were approximately 5%-15% lower than those of controls after week 70. Survival of rats at 104 weeks was as follows: male: control, 18/50; 2 ppm, 17/50; 5 ppm, 4/50; female: 25/50; 34/50; 15/50; survival of rats at the top concentration was reduced due to neoplasia. Mean body weights of exposed mice were variable and ranged as much as 10% below those of controls during the second year of the studies. Survival of exposed male mice at 104 weeks was significantly lower than that of controls due to neoplasia (control, 37/50; 0.5 ppm, 26/50; 2 ppm, 15/50). Survival of female mice was not significantly affected by exposure to tetranitromethane (31/50; 28/50; 24/50). Neoplastic and Nonneoplastic Effects in the Two-Year Studies: Effects of exposure to tetranitromethane were limited to the respiratory tract. Hyperplasia of the alveolar and bronchiolar epithelium was observed at increased incidences in exposed rats. The incidence of alveolar/bronchiolar adenomas and carcinomas were markedly increased in exposed male and female rats, with carcinomas (many of which metastasized to other sites) occurring in nearly all rats exposed to the top concentration of 5 ppm (adenomas or carcinomas-- male: control, 1/50; 2 ppm, 33/50; 5 ppm, 46/50; female: 0/50; 22/50; 50/50). Many of the rats exposed to 5 ppm also had squamous cell carcinomas of the lung (male: 0/50; 1/50; 19/50; female: 0/50; 1/50; 12/50). Hyperplasia of the respiratory epithelium and chronic inflammation of the nasal mucosa were observed at increased incidences in exposed male and female rats. Squamous metaplasia of the respiratory epithelium was increased in exposed male rats. No neoplasms of the nasal passage were seen. In exposed mice, hyperplasia of the alveolar and br were seen. In exposed mice, hyperplasia of the alveolar and bronchiolar epithelium was observed at increased incidences. Alveolar/bronchiolar neoplasms, primarily carcinomas (many of which metastasized to other sites), were increased in exposed male and female mice (male: control, 12/50; 0.5 ppm, 27/50; 2 ppm, 47/50; female: 4/49; 24/50; 49/50). Chronic inflammation of the nasal mucosa and hyperplasia and squamous metaplasia of the respiratory epithelium of the nasal cavity occurred at increased incidences in female mice exposed to 2 ppm. No primary neoplasms of the nasal passage were observed in mice. Oncogene Analysis: DNA from 14/19 rat and 4/4 mouse lung neoplasms caused morphologic transformation after transfection into cultured NIH/3T3 fibroblasts. The transforming gene from both rat and mouse lung neoplasms was determined by Southern blot analysis to be an activated K-ras oncogene. Further studies showed a GC-->AT transition in the second base of the 12th codon of the K-ras oncogene. Genetic Toxicology: Tetranitromethane was mutagenic in S. typhimurium strains TA98, TA100, and TA1535 with and without exogenous metabolic activation (S9); no mutagenic activity was observed in TA1537 with or without S9. Chromosomal aberrations were observed in CHO cells treated in vitro with tetranitromethane in the presence of S9. Sister chromatid exchanges were induced in CHO cells in the absence of S9. Conclusions: Under the conditions of these 2 year inhalation studies, there was clear evidence of carcinogenic activity of tetranitromethane for male and female F344/N rats and male and female B6C3F1 mice, based on increased incidences of alveolar/bronchiolar neoplasms in both species and squamous cell carcinomas of the lung in rats. Chronic inflammation of the nasal mucosa was related to exposure in rats and female mice, and hyperplasia and squamous metaplasia of the respiratory epithelium were increased in exposed male rats. Synonym: TNM PMID- 12637974 TI - NTP toxicology and carcinogenesiss studies of citral (microencapsulated) (CAS No. 5392-40-5) in F344/N rats and B6C3F1 mice (feed studies). AB - Citral is used primarily as lemon flavoring in foods, beverages, and candies. It is also used as a lemon fragrance in detergents, perfumes, and other toiletries. Citral was nominated by the National Cancer Institute for study because of its widespread use in foods, beverages, cosmetics, and other consumer products and its structure as a representative beta-substituted vinyl aldehyde. Male and female F344/N rats and B6C3F1 mice were exposed to microencapsulated citral (greater than 96% pure) in feed for 14 weeks or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, mouse bone marrow cells, and mouse peripheral blood erythrocytes. 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female F344/N rats were fed diets containing starch microcapsules with a load of 31.3% citral. The concentration of citral in the diet was 3,900, 7,800, 15,600, or 31,300 ppm microencapsulated citral (equivalent to average daily doses of approximately 345, 820, 1,785, and 1,585 mg citral/kg body weight to males and 335, 675, 1,330, and 2,125 mg/kg to females) for 14 weeks. Additional groups of 10 male and 10 female rats received untreated feed (untreated controls) or feed containing placebo microcapsules (vehicle controls). In the second week of the study, all rats in the 31,300 ppm groups were killed moribund. Mean body weights of exposed males and females that survived to the end of the study were generally significantly less than those of the vehicle controls. Feed consumption by 15,600 and 31,300 ppm males and females was less than that by the vehicle controls during the first week of the study. Males and females in the 31,300 ppm groups exhibited listlessness, hunched posture, absent or slow paw reflex, and dull eyes. Exposure of rats to citral may have been associated with forestomach epithelial hyperplasia and hyperkeratosis, bone marrow atrophy and hemorrhage, and nephrotoxicity. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice were fed diets containing 3,900, 7,800, 15,600, or 31,300 ppm microencapsulated citral (equivalent to average daily doses of approximately 745, 1,840, 3,915, and 8,110 mg/kg to males and 790, 1,820, 3,870, and 7,550 mg/kg to females) for 14 weeks. Additional groups of 10 male and 10 female mice received untreated feed (untreated controls) or feed containing placebo microcapsules (vehicle controls). In the second week of the study, four males in the 31,300 ppm group were killed moribund. Mean body weights of all exposed groups of males and females were significantly less than those of the vehicle controls. Feed consumption by females exposed to 7,800 ppm or greater was less than that by the vehicle controls during the first week of the study. By the end of the study, feed consumption by all exposed groups was greater than that by the vehicle controls. Mice in the 15,600 and 31,300 ppm groups were generally thin and lethargic; a few males in the 7,800 ppm group were also thin. The incidences of ovarian atrophy were significantly increased in females exposed to 15,600 or 31,300 ppm. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female F344/N rats were fed diets containing 1,000, 2,000, or 4,000 ppm microencapsulated citral for 2 years. Additional groups of 50 male and 50 female rats received untreated feed (untreated controls) or feed containing placebo microcapsules (vehicle controls). Dietary concentrations of 1,000, 2,000, and 4,000 ppm delivered average daily doses of approximately 50, 100, and 210 mg/kg to males and females. Survival of all exposed groups of males was significantly greater than that of the vehicle control group. Mean body weights of rats exposed to 4,000 ppm were generally less than those of the vehicle controls from week 49 (males) or 25 (females) to the end of the study. Feed consumption by exposed groups was similar to that by the vehicle controls. No neoplasms or nonneoplastic lesions were attributed to exposure to citral. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female B6C3F1 mice were fed diets containing 500, 1,000, or 2,000 ppm microencapsulated citral for 2 years. Additional groups of 50 male and 50 female mice received untreated feed (untreated controls) or feed containing placebo microcapsules (vehicle controls). Dietary concentrations of 500, 1,000, and 2,000 ppm delivered average daily doses of approximately 60, 120, and 260 mg/kg to males and females. Survival of exposed males and females was similar to that of the vehicle control groups. Mean body weights of mice exposed to 1,000 or 2,000 ppm were generally less than those of the vehicle controls throughout the study, and mean body weights of 500 ppm females were less from week 30 to the end of the study. Feed consumption by the exposed groups was similar to that by the vehicle controls. The incidences of malignant lymphoma occurred with a positive trend in female mice, and the incidence in 2,000 ppm females was significantly greater than that in the vehicle control group. Tissues most commonly affected by malignant lymphoma were the spleen, mesenteric lymph node, thymus, and, to a lesser extent, the ovary. GENETIC TOXICOLOGY: Citral was not mutagenic in S. typhimurium strain TA98, TA100, TA1535, or TA1537 with or without induced rat or hamster liver S9 enzymes. In cytogenetic tests with cultured Chinese hamster ovary cells, citral induced sister chromatid exchanges with and without S9, but chromosomal aberrations were not significantly increased after exposure to citral, with or without S9. Negative results were obtained in an in vivo bone marrow micronucleus test in male B6C3F1 mice treated by intraperitoneal injection with 250 to 750 mg/kg daily for 3 days. Likewise, no increases in the frequencies of micronucleated erythrocytes were observed in peripheral blood samples collected from male and female mice within 24 hours of the final exposure in the 14-week study. In conclusion, citral gave negative results in in vitro and in vivo tests for genotoxicity, with the exception of the in vitro mammalian cell test for sister chromatid exchange induction CONCLUSIONS: Under the conditions of these 2-year feed studies, there was no evidence of carcinogenic activity of citral in male or female F344/N rats exposed to 1,000, 2,000, or 4,000 ppm. There was no evidence of carcinogenic activity of citral in male B6C3F1 mice exposed to 500, 1,000, or 2,000 ppm. There was equivocal evidence of carcinogenic activity in female B6C3F1 mice based on increased incidences of malignant lymphoma. PMID- 12637975 TI - Methanol utilization by a novel thermophilic homoacetogenic bacterium, Moorella mulderi sp. nov., isolated from a bioreactor. AB - A thermophilic, anaerobic, spore-forming bacterium (strain TMS) was isolated from a thermophilic bioreactor operated at 65 degrees C with methanol as the energy source. Cells were gram-positive straight rods, 0.4-0.6 microm x 2-8 microm, growing as single cells or in pairs. The temperature range for growth was 40-70 degrees C with an optimum at 65 degrees C. Growth was observed from pH 5.5 to 8.5, and the optimum pH was around 7. The salinity range for growth was 0-45 g NaCl l(-1 )with an optimum at 10 g l(-1). The isolate was able to grow on methanol, H(2)-CO(2 )(80/20%, v/v), formate, lactate, pyruvate, glucose, fructose, cellobiose and pectin. The bacterium reduced thiosulfate to sulfide. The G+C content of the DNA was 53 mol%. Comparison of 16S rRNA genes revealed that strain TMS is related to Moorella glycerini (96%, sequence similarity), Moorella thermoacetica (92%) and Moorella thermoautotrophica (92%). On the basis of physiological and phylogenetic differences, strain TMS is proposed as a new species within the genus Moorella, Moorella mulderi sp. nov. (=DSM 14980, =ATCC BAA-608). PMID- 12637976 TI - Reversible spongiform leucoencephalopathy after inhalation of heated heroin. AB - OBJECTIVE: To describe the clinical course and imaging findings in a young man who developed a spongiform leucoencephalopathy from heroin-vapour inhalation, and to discuss the treatments which may have contributed to the unexpected favourable outcome in this case. DESIGN: Case report. SETTING: Intensive care unit of a university teaching hospital. PATIENT: A patient who developed a near fatal toxic leucoencephalopathy with impressive clinical recovery and reversible white matter changes on imaging. MEASUREMENTS AND RESULTS: Successive computed tomography scans and magnetic resonance imaging over 7 months showed evolution from bilateral extensive involvement of the cerebral white matter to almost complete resolution accompanied by the development of periventricular lesions suggestive of necrosis. Despite the fact that the patient had stretching spasms for several days, the outcome was favourable with prolonged supportive care and antioxidant therapy by ubiquinone (coenzyme Q). CONCLUSION: This case demonstrates that prolonged intensive care is of paramount importance in patients with spongiform leucoencephalopathy after inhalation of heated heroin, that abnormalities of cerebral white matter may be slowly regressive, and supports the use of coenzyme Q in severe forms of the disease. PMID- 12637978 TI - Proinsulin and acute insulin response independently predict Type 2 diabetes mellitus in men--report from 27 years of follow-up study. AB - AIMS/HYPOTHESIS: Defects in insulin secretion and insulin action, resulting in compensatory hyperinsulinaemia, are the major abnormalities in the development of Type 2 diabetes mellitus (Type 2 diabetes). The most frequently used conventional immunoreactive assays for insulin cross-react with proinsulin. In short-term studies (<5 years), proinsulin predicts the development of Type 2 diabetes. We studied, with a 27-year follow-up, the longitudinal relationships between intact proinsulin, 32-33 split proinsulin, specific and immunoreactive insulin (IRI), acute insulin response (AIR) after an intravenous glucose load and the development of Type 2 diabetes in a population-based cohort of 50-year-old men. METHODS: Fasting peptide concentrations were measured in plasma samples, stored since 1970-73 using specific two-site immunometric assays. IRI was measured at baseline using radioimmunoassay. Associations between development of Type 2 diabetes and predictor variables, were analysed with logistic regression. Results are shown as odds ratios (ORs) with their 95% confidence intervals (CIs) for a one standard deviation increase in a predictor variable. RESULTS: Cumulative incidence of Type 2 diabetes was 33% over 27 years of follow-up. Intact proinsulin (OR, 1.57, CI, 1.16-2.14), and 32-33 split proinsulin (OR, 1.70, CI, 1.20-2.39) were associated with development of Type 2 diabetes, independent of AIR, adjusted for BMI and fasting glucose, whereas specific insulin was not (OR, 1.31, CI, 0.98-1.77), nor was IRI (OR, 1.25, CI, 0.96-1.63). Proinsulin and AIR interacted in the development of Type 2 diabetes (p<0.05). CONCLUSION/INTERPRETATION: Proinsulin predicts the development of Type 2 diabetes mellitus over a 27-year period. PMID- 12637979 TI - Evaluation of granulocyte-colony stimulating factor (Filgrastim) in infected diabetic foot ulcers. AB - AIMS/HYPOTHESIS: To re-evaluate the use of Granulocyte-Colony Stimulating Factor (G-CSF) in the treatment of infected diabetic foot ulcers. METHODS: Thirty-seven diabetic subjects were randomised to Granulocyte-Colony Stimulating Factor (G CSF) (n=20) or placebo (n=17). The primary endpoint was resolution of cellulitis, which was evaluated clinically and with an infection summary score. Patients were hospitalised for 10 days and received subcutaneously either 5 microg/kg G-CSF or placebo daily. Ulcers were treated with a standard wound protocol and the patients were instructed to stay in bed. All subjects received antibiotics (clindamycin and ciprofloxacin) intravenously until the inflammation had subsided. RESULTS: Patients who received G-CSF did not have an earlier resolution of clinically defined cellulitis (p=0.57). The infection summary score declined, but comparably, in both groups (G-CSF: 29.5+/-18.4 to 6.7+/-6.3 p<0.001, placebo: 24.2+/-16.9 to 8.9+/-7.2 p<0.001). The ulcer volume, which was not greater among placebo patients, was reduced by 59% in G-CSF and by 35% in placebo patients. CONCLUSION/INTERPRETATION: We conclude that antibiotic and non weight-bearing therapy (bed rest) accelerated the resolution of cellulitis in infected foot ulcers. Additional treatment with G-CSF had no further beneficial effect. PMID- 12637980 TI - The CAG repeat polymorphism in the androgen receptor gene modulates body fat mass and serum concentrations of leptin and insulin in men. AB - AIMS/HYPOTHESIS: The relationship of androgens to the metabolic syndrome has not been resolved. The polymorphic number of CAG repeats within the androgen receptor gene is inversely associated with the transcriptional activity of target genes. This polymorphism might thus influence testosterone effects on body fat content and serum concentrations of leptin and insulin. The direct and indirect role of androgens within the metabolic syndrome should become clearer if this genetically determined effector is taken into account. METHODS: The hypothesis was investigated in a cross-sectional study involving 106 healthy 20-50 year old males. RESULTS: Multiple regression models showed a positive independent correlation of the CAG repeat number with body fat content, leptin and insulin (partial r=0.39, 0.36 and 0.28, p<0.001, p<0.001 and p=0.006, respectively). Factor analysis yielded a five-dimensional model: two dimensions were influenced by the androgen receptor polymorphism, namely "body composition" which consisted of leptin, body fat mass, insulin, the number of CAG repeats (positive loadings) and physical activity (negative loading), and "lipid profile" which comprised low density lipoprotein cholesterol, cigarette smoking, triglycerides (positive loadings) as well as high density lipoprotein cholesterol and number of CAG repeats (negative loadings). CONCLUSIONS/INTERPRETATION: A low number of CAG repeats were independently associated with protective parameters (low body fat mass and plasma insulin) as well as with adverse parameters (low high density lipoprotein cholesterol concentrations). This suggests that the pivotal role of this polymorphism in modulating androgen effects on cardiovascular risk factors is of a complex nature and implies that its clinical impact, similar to that of androgens, is dependent on exogenous cofactors. PMID- 12637977 TI - The relative contributions of insulin resistance and beta-cell dysfunction to the pathophysiology of Type 2 diabetes. AB - The relative contributions of insulin resistance and beta-cell dysfunction to the pathophysiology of Type 2 diabetes have been debated extensively. The concept that a feedback loop governs the interaction of the insulin-sensitive tissues and the beta cell as well as the elucidation of the hyperbolic relationship between insulin sensitivity and insulin secretion explains why insulin-resistant subjects exhibit markedly increased insulin responses while those who are insulin sensitive have low responses. Consideration of this hyperbolic relationship has helped identify the critical role of beta-cell dysfunction in the development of Type 2 diabetes and the demonstration of reduced beta-cell function in high risk subjects. Furthermore, assessments in a number of ethnic groups emphasise that beta-cell function is a major determinant of oral glucose tolerance in subjects with normal and reduced glucose tolerance and that in all populations the progression from normal to impaired glucose tolerance and subsequently to Type 2 diabetes is associated with declining insulin sensitivity and beta-cell function. The genetic and molecular basis for these reductions in insulin sensitivity and beta-cell function are not fully understood but it does seem that body-fat distribution and especially intra-abdominal fat are major determinants of insulin resistance while reductions in beta-cell mass contribute to beta-cell dysfunction. Based on our greater understanding of the relative roles of insulin resistance and beta-cell dysfunction in Type 2 diabetes, we can anticipate advances in the identification of genes contributing to the development of the disease as well as approaches to the treatment and prevention of Type 2 diabetes. PMID- 12637982 TI - Thiazolidinediones (PPARgamma ligands) increase IRS-1, UCP-2 and C/EBPalpha expression, but not transdifferentiation, in L6 muscle cells. AB - AIMS/HYPOTHESIS: Several effects of thiazolidinediones (TZD) have been shown in adipose tissue but very little is known about the effects in skeletal muscle. We examined the effects of TZD and PPARalpha ligands on the expression of different genes in L6 muscle cells. METHODS: L6 muscle cells were exposed to PPARalpha and PPARgamma ligands for different times. The gene expression of the signalling molecules IRS-1, IRS-2, PKB/Akt, the transcription factor C/EBPalpha, the uncoupling protein UCP-2 and the adipocyte marker aP2 were measured with real time PCR. To directly examine the effect of C/EBPalpha on gene expression, we also transfected L6 cells with this gene. RESULTS: L6 muscle cells showed a low expression of PPARgamma 1, no expression of PPARgamma 2, and this was not changed by TZD. PPARgamma, but not PPARalpha, ligands rapidly increased the expression of C/EBPalpha while UCP-2 and in particular the IRS-1 gene was activated with a slow onset (12-24 h). In contrast, neither PKB/Akt nor IRS-2 expression were changed. Transfection with C/EBPalpha did not increase IRS-1 expression. There was no evidence of transdifferentiation of the muscle to an adipocyte phenotype by TZD since no aP2 expression was found. CONCLUSION/INTERPRETATION: TZD increase IRS-1, UCP2 and C/EBPalpha expression in L6 muscle cells. Activation of C/EBPalpha by TZD could be necessary but it is not sufficient to account for the increased IRS 1 expression. PMID- 12637981 TI - Sympathetic mediated vasomotion and skin capillary permeability in diabetic patients with peripheral neuropathy. AB - AIMS/HYPOTHESIS: A loss of sympathetic function could lead to changes in capillary fluid filtration in diabetic patients. We investigated whether a decreased sympathetically mediated vasomotion in the skin in diabetic patients with peripheral neuropathy is associated with an abnormal capillary leakage. METHODS: Three matched groups were studied: 18 diabetic patients with documented peripheral neuropathy (DN), 18 diabetic patients without peripheral neuropathy (D), and 18 healthy control subjects (C). Sensory and motor nerve function of the distal extremities were assessed by standard neurography, and expressed in a sensory-motor nerve function score. Sympathetic vasomotion of the skin microcirculation was assessed by determining the power of blood flow variability in the low-frequency (0.02-0.14 Hz) band by spectral analysis of laser Doppler flowmetry at the median ankle. Skin capillary leakage was evaluated by sodium fluorescein videodensitometry at the same site of the foot. RESULTS: Sympathetically mediated vasomotion of the foot skin microcirculation was lower in diabetic patients with documented peripheral neuropathy compared with diabetic patients without peripheral neuropathy and control subjects (p<0.001). Capillary sodium fluorescein leakage was larger in 18 diabetic patients with documented peripheral neuropathy than in diabetic patients without peripheral neuropathy (p<0.02) and C (p<0.005). Multiple regression analysis disclosed that a reduced sympathetically mediated vasomotion, together with a lower sensory-motor nerve function score, independently contributed to the variance in sodium fluorescein leakage, for 30% (p<0.001) and 17% (p<0.01), respectively. CONCLUSIONS: A loss of sympathetic tone, apart from sensory-motor nerve dysfunction, seems to be a major determinant of an increased capillary permeability in diabetic patients with neuropathy. PMID- 12637983 TI - Rate of fall of blood glucose and physiological responses of counterregulatory hormones, clinical symptoms and cognitive function to hypoglycaemia in Type I diabetes mellitus in the postprandial state. AB - AIMS/HYPOTHESIS: The aim of this study was to establish the effect of a rate of decreasing plasma glucose concentrations on responses to hypoglycaemia, i.e. release of counterregulatory hormones, perception of symptoms, deterioration of cognitive function, and rates of forearm noradrenaline spillover, in the postprandial condition and in the sitting position. METHODS: We studied 11 subjects with Type I (insulin-dependent) diabetes mellitus, twice during clamped insulin-induced hypoglycaemia (2.4 mmol/l) after eating in the sitting position. On one occasion, plasma glucose was decreased at the rate of 0.1+/-0.003 mmol x min(-1) x l(-1) (fast fall), on the other at the rate of 0.03+/-0.001 mmol x min( 1) x l(-1) (slow fall). Subjects underwent a control euglycaemic clamp study as well. RESULTS: In response to fast-fall as compared to slow-fall hypoglycaemia, which was about 30 min longer, cognitive tasks were performed as follows: Trail Making B, PASAT 2 s, Digit Vigilance Test and Verbal Memory deteriorated more, adrenaline increased less (2.8+/-0.5 vs 3.5+/-0.7 nmol/l, p=0.03), forearm noradrenaline spillover was greater (6.5+/-1.0 vs 5.2+/-0.4 pmol x min(-1) x 100 ml(-1), p=0.04), and symptoms were no different. After recovery from hypoglycaemia, cognitive function was still deteriorated compared to the baseline with no difference between fast and slow-fall hypoglycaemia. The evident response of glucagon to postprandial hypoglycaemia contrasted with the blunted or absent response in the fasting state. CONCLUSION/INTERPRETATION: In the postprandial condition and sitting position, fast-fall hypoglycaemia is more dangerous than slow-fall, because it deteriorates cognitive function more, and activates responses of counterregulatory hormones less than slow-fall hypoglycaemia. PMID- 12637984 TI - Signs of beta-cell autoimmunity and HLA-defined diabetes susceptibility in the Finnish population: the sib cohort from the Type 1 Diabetes Prediction and Prevention Study. AB - AIMS/HYPOTHESIS: To assess the role of HLA-defined genetic diabetes susceptibility in the appearance of signs of beta-cell autoimmunity in a series of children derived from the general population. METHODS: Tests for five HLA DQB1 alleles and four diabetes-associated autoantibodies were carried out on 1,584 older sibs of infants with an increased HLA-defined genetic risk of Type 1 diabetes. The DQB1 genotypes were classified into those conferring high (* 02/0302), moderate (* 0302/x; where x indicates * 0302 or a non-defined allele), low (* 0301/0302, * 02/0301, * 02/x, * 0302/0602, * 0302/0603; where x indicates * 02 or a non-defined allele) or decreased risk (other genotypes). RESULTS: Both islet cell antibodies (ICA) and GAD65 antibodies (GADA) were more frequent among the sibs with the high-risk genotype than among those with a low or decreased risk. Insulin autoantibodies and IA-2 antibodies (IA-2A) were more prevalent in the high-risk than low-risk sibs. Sibs with moderate-risk genotypes tested positive for ICA, GADA and IA-2A more often than sibs with genotypes conferring decreased risk. Autoantibody titres were also dependent on the genetic risk with high risk sibs having the highest values. Sibs carrying high-risk or moderate risk genotypes tested positive for multiple antibodies (> or =2) more often than did the sibs with low or decreased genetic risk. CONCLUSIONS/INTERPRETATION: The data show that HLA-defined susceptibility to Type 1 diabetes has an effect on both the quality and quantity of humoral beta-cell autoimmunity in unaffected children derived from the general population. PMID- 12637985 TI - The effect of apolipoprotein E deficiency on islet amyloid deposition in human islet amyloid polypeptide transgenic mice. AB - AIMS/HYPOTHESIS: Islet amyloid deposits are present in over 85% of Type 2 diabetic patients and have been suggested to be pathogenic. The mechanism that converts islet amyloid polypeptide (IAPP), the unique component of these deposits, into amyloid fibrils in vivo is not known. The amino acid sequence of IAPP is critical but insufficient for beta-pleated sheet formation. As apolipoprotein E (apoE), another component of islet amyloid deposits, plays a critical role in amyloid formation in Alzheimer's disease, we hypothesised that apoE could play an important role in islet amyloid formation. METHODS: Transgenic mice expressing the human form of IAPP ( hIAPP (+/0)) were crossbred with apoE deficient ( apoE (-/-)) mice and followed for 12 months, at which time the prevalence and severity of islet amyloid, as well as plasma glucose, hIAPP, immunoreactive insulin (IRI) and lipid concentrations were measured. RESULTS: The prevalence and severity of islet amyloid after one year of follow up were comparable among hIAPP (+/0) mice that were apoE (+/+), apoE (+/-) or apoE (-/-). Differences in glucose tolerance, lipid abnormalities or changes in pancreatic content or plasma concentrations of hIAPP and/or IRI did not account for these findings. CONCLUSION/INTERPRETATION: Our data shows that, unlike in the localized amyloidosis in the brain characteristic of Alzheimer's disease, apoE is not critical for islet amyloid formation in a transgenic mouse model of Type 2 diabetes mellitus. These results indicate that the mechanisms of localised amyloid formation probably vary among different amyloid-associated disorders. Therefore, therapeutic strategies targeting apoE might not apply equally to patients with different amyloid associated diseases. PMID- 12637986 TI - Protection of rat pancreatic islets by potassium channel openers against alloxan, sodium nitroprusside and interleukin-1beta mediated suppression--possible involvement of the mitochondrial membrane potential. AB - AIMS/HYPOTHESIS: We aimed to study the effects of two K(ATP) channel openers (KCO), diazoxide and the more potent compound NNC 55-0118, on beta-cell suppression and/or toxicity induced by alloxan, sodium nitroprusside and IL 1beta. METHODS: Islets from rats were exposed to 0.3 mmol/l diazoxide or NNC 55 0118 for 30 min and either alloxan (0.5 mmol/l), sodium nitroprusside (0.5 mmol/l) or IL-1beta (12.5 or 25 U/ml) were added and the incubation continued for 30 min. Islets were then washed and incubated for 24 h before examination. RESULTS: After exposure to alloxan, islets showed reduced glucose oxidation rate and impaired glucose-stimulated insulin release. NNC 55-0118 counteracted the effects of alloxan, while diazoxide was less effective. After treatment with sodium nitroprusside, islet glucose oxidation rates were reduced and this was prevented by pretreatment with NNC 55-0118. In short-term experiments the potassium channel openers (KCOs) did not influence the IL-1beta effect on insulin secretion. However, long-term addition (24 h) of NNC 55-0118 counteracted IL 1beta induced inhibition of the glucose oxidation rate. It was shown, using the fluorescent probe JC-1, that the mitochondrial membrane potential was reduced by the potassium channel openers (KCOs), most strongly by NNC 55-0118. Nevertheless culture with KCOs for 72 h did not cause irreversible damage to the islets. CONCLUSION/INTERPRETATION: Potassium channel openers (KCOs), in particular NNC 55 0118, prevented the toxic effects of alloxan and sodium nitroprusside. IL-1beta mediated suppression was reduced by NNC 55-0118 provided the long-term addition of the potassium channel opener (KCO). The protective mechanism of potassium channel openers (KCOs) might involve a decrease of the mitochondrial membrane potential. PMID- 12637987 TI - Prediction of severe hypoglycaemia by angiotensin-converting enzyme activity and genotype in type 1 diabetes. AB - AIMS/HYPOTHESIS: We have previously shown a strong relationship between high angiotensin-converting enzyme (ACE) activity, presence of the deletion (D) allele of the ACEgene and recall of severe hypoglycaemic events in patients with Type 1 diabetes. This study was carried out to assess this relationship prospectively. METHODS: We followed 171 adult outpatients with Type 1 diabetes in a one-year observational study with the recording of severe hypoglycaemia. Participants were characterised by serum ACE activity and ACE genotype and not treated with ACE inhibitors or angiotensin II receptor antagonists. RESULTS: There was a positive relationship between serum ACE activity and rate of severe hypoglycaemia with a 2.7 times higher rate in the fourth quartile of ACE activity compared to the first quartile (p=0.0007). A similar relationship was observed for the subset of episodes with coma (2.9 times higher rate in fourth quartile compared to first quartile; p=0.048). The impact of serum ACE activity was most pronounced in C peptide negative subjects (4.2 times higher rate in fourth quartile compared to first quartile; p=0.003), and in this subgroup carriers of the D allele of the ACEgene had higher rates of severe hypoglycaemia compared to the group homozygous for the insertion (I) allele. In a multiple regression analysis high serum ACE activity and impaired awareness of hypoglycaemia were identified as the only significant predictors of severe hypoglycemia. CONCLUSION: High ACE activity and the presence of the D allele of the ACE gene predict a high rate of severe hypoglycaemia in Type 1 diabetes. PMID- 12637988 TI - Effects of acute hypoglycaemia on auditory information processing in adults with Type I diabetes. AB - AIMS/HYPOTHESIS: Acute hypoglycaemia in humans causes general impairment of cognitive function, but information about its effects on more specific cognitive processes is limited. METHODS: Basic aspects of auditory function were studied in 15 adults with uncomplicated Type I (insulin-dependent) diabetes mellitus. Two separate hyperinsulinaemic glucose clamp procedures were done on different study days, in a counterbalanced fashion, either maintaining euglycaemia (blood glucose 5.0 mmol x l(-1)) or inducing hypoglycaemia (blood glucose 2.6 mmol x l(-1)). During each study, the subjects performed a battery of auditory and cognitive function tasks. RESULTS: Hypoglycaemia caused deterioration in mental efficiency as assessed by Digit Symbol (p<0.001) and Trail Making B (p=0.004) tasks. Acute hypoglycaemia also caused deterioration in one of three measures of simple auditory processing (single-tone loudness, p=0.001) and in auditory temporal processing (p=0.007). The amplitude and latency of auditory N100, P200 and P300 event-related potentials were not affected, but the amplitude of the N240 potential was reduced during acute hypoglycaemia. CONCLUSION/INTERPRETATION: Our findings are consistent with other recognised disruptive effects of acute hypoglycaemia on sensory information processing in non-diabetic and diabetic adults, including adverse effects on auditory information processing in non diabetic subjects. These derangements have implications for the everyday activities of people with Type I diabetes who are frequently exposed to acute hypoglycaemia. PMID- 12637989 TI - Interaction of glucose and long chain fatty acids (C18) on antioxidant defences and free radical damage in porcine vascular smooth muscle cells in vitro. AB - AIMS/HYPOTHESIS: Abnormalities of glucose and fatty acid metabolism in diabetes are believed to contribute to the development of oxidative stress and the long term vascular complications of the disease; therefore the interactions of glucose and long chain fatty acids on free radical damage and endogenous antioxidant defences were investigated in vascular smooth muscle cells. METHODS: Porcine vascular smooth muscle cells were cultured in 5 mmol/l or 25 mmol/l glucose for 10 days. Fatty acids, stearic acid (18:0), oleic acid (18:1), linoleic acid (18:2) and alpha-linolenic acid (18:3) were added with defatted bovine serum albumin as a carrier for the final three days. RESULTS: Glucose (25 mmol/l) alone caused oxidative stress in the cells as evidenced by free radical-mediated damage to DNA, lipids, and proteins. The addition of fatty acids (0.2 mmol/l) altered the profile of free radical damage; the response was J-shaped with respect to the degree of unsaturation of each acid, and oleic acid was associated with least damage. At a lower concentration alpha-linolenic acid (0.01 mmol/l) was markedly different in that, when added to 25 mmol/l glucose it resulted in a decrease in free radical damage to DNA, lipids and proteins. This was accompanied by a marked increase in antioxidant and glutathione concentrations as well as by increased gene expression is of gamma-glutamylcysteine synthetase, the rate-limiting enzyme in glutathione synthesis. CONCLUSIONS/INTERPRETATION: The results clearly show that glucose and fatty acids interact in the production of oxidative stress in vascular smooth muscle cells. PMID- 12637990 TI - HGF/NK4 inhibited VEGF-induced angiogenesis in in vitro cultured endothelial cells and in vivo rabbit model. AB - AIMS/HYPOTHESIS: As vascular endothelial growth factor (VEGF) plays a pivotal role in the development of diabetic retinopathy, inhibition of angiogenesis induced by VEGF is crucial to treat diabetic retinopathy. HGF (hepatocyte growth factor)/NK4, containing the N-terminal hairpin domain and the four subsequent kringle domains of HGF, is considered as a specific antagonist for HGF. Our aim was to explore the inhibitory effects of HGF/NK4 on angiogenesis induced by VEGF. METHODS: To analyze the in vivo angiogenesis, we used rabbit corneal micropocket assay. Proliferation and migration of human endothelial cells, expression of ets 1, an essential transcription factor for angiogenesis, and the phosphorylation of extracellular signal-regulated kinase (ERK) was examined with or without HGF/NK4. RESULTS: Using corneal micropocket assay, in vivo administration of HGF/NK4 inhibited angiogenesis induced by VEGF. HGF/NK4 inhibited proliferation and migration of human endothelial cells induced by VEGF in a dose-dependent manner. Interestingly, VEGF-mediated phosphorylation of ERK was significantly attenuated by HGF/NK4. Of importance, HGF/NK4 attenuated the increase in ets-1 protein stimulated by VEGF. Nevertheless, HGF/NK4 did not affect phosphorylation of VEGF receptor-2 [kinase domain region (KDR)/foetal liver kinase (Flk)-1]. Although tyrosine phosphatase inhibitor (Na(3)VO(4)), or okadaic acid, serine-threonin kinase inhibitor, did not prevent the inhibition of ERK phosphorylation by HGF/NK4, co-incubation of HGF/NK4 with VEGF significantly diminished mitogen activated protein (MAP) ERK kinase (MEK) phosphorylation (p<0.01). CONCLUSIONS/INTERPRETATION: Overall, HGF/NK4 inhibited angiogenesis induced by VEGF through inhibition of phosphorylation of ERK and ets-1 expression in in vitro cultured endothelial cells and in vivo rabbit model. PMID- 12637992 TI - To: Saleh MC, Wheeler MB, Chan CB (2002) Uncoupling protein-2: evidence for its function as a metabolic regulator. Diabetologia 45:174-187. PMID- 12637991 TI - C-peptide stimulates Na+,K+-ATPase activity via PKC alpha in rat medullary thick ascending limb. AB - AIMS/HYPOTHESIS: C-peptide, the cleavage product of proinsulin processing exerts physiological effects including stimulation of Na(+),K(+)-ATPase in erythrocytes and renal proximal tubules. This study was undertaken to assess the physiological effects of connecting peptide on Na(+),K(+)-ATPase activity in the medullary thick ascending limb of Henle's loop. METHODS: Na(+),K(+)-ATPase activity was measured as the ouabain-sensitive generation of (32)Pi from gamma[(32)P]-ATP and (86)Rb uptake on isolated rat medullary thick ascending limbs. The cell-surface expression of Na(+),K(+)-ATPase was evaluated by Western blotting of biotinylated proteins, and its phosphorylation amount was measured by autoradiography. The membrane-associated fraction of protein kinase C isoforms was evaluated by Western blotting. RESULTS: Rat connecting peptide concentration-dependently stimulated Na(+),K(+)-ATPase activity with a threshold at 10(-9) mol/l and a maximal effect at 10(-7) mol/l. C-peptide (10(-7) mol/l) already stimulates Na(+),K(+)-ATPase activity after 5 min with a plateau from 15 to 60 min. C peptide (10(-7) mol/l) stimulated Na(+),K(+)-ATPase activity and (86)Rb uptake to the same extent, but did not alter Na(+),K(+)-ATPase cell surface expression. The stimulation of Na(+),K(+)-ATPase activity was associated with an increase in Na(+),K(+)-ATPase alpha-subunit phosphorylation and both effects were abolished by a specific protein kinase C inhibitor. Furthermore, connecting peptide induced selective membrane translocation of PKC-alpha. CONCLUSION/INTERPRETATION: This study provides evidence that in rat medullary thick ascending limb, C-peptide stimulates Na(+),K(+)-ATPase activity within a physiological concentration range. This effect is due to an increase in Na(+),K(+)-ATPase turnover rate that is most likely mediated by protein kinase C-alpha phosphorylation of the Na(+),K(+) ATPase alpha-subunit, suggesting that C-peptide could control Na(+) reabsorption during non-fasting periods. PMID- 12638006 TI - Fold prediction and comparative modeling of Bdm1: a probable alpha/beta hydrolase associated with hot water epilepsy. AB - Hot water epilepsy (HWE) is a benign and rare form of reflex epilepsy that occurs most commonly in humans. Bdm1 is one of the proteins whose mRNA transcript is overexpressed during HWE in a rat model. We show, by sequence analysis and fold recognition methods, that Bdm1 has strong structural similarities to alpha/beta hydrolases like the thioesterases. A three-dimensional model derived by comparative modeling methods allowed the search for catalytic residues using a flexible functional template characteristic of these enzymes. We predict that Bdm1 might be regulated by homocysteine levels by means of direct participation in degradation pathways. PMID- 12638007 TI - Comparing the modeled structures of PR-4 proteins from wheat. AB - We have constructed three-dimensional models of four pathogenesis-related (PR) proteins from wheat (wheatwins) belonging to the PR-4 family. All the models were based on the knowledge of the tertiary structure of barwin, a highly homologous protein from barley. Wheatwin1 and wheatwin2 differ in two amino acid residues (positions 62 and 68) out of 125. Wheatwin4 differs from wheatwin2 in one residue at position 78, while wheatwin3 differs from wheatwin1 in one residue at position 88. The global folding and the secondary structures were very similar through all the sequences, including the regions of the amino acid substitutions. The main differences were found in the traits 15-21, 84-86 and 91-93. Trait 15-21 was predicted as ss-sheet in wheatwin4 and random-coil in the other proteins. Trait 84-86 was predicted as ss-sheet in wheatwin3 and random-coil in the other proteins. Trait 91-93 was predicted as random coil in wheatwin1 and wheatwin3 and ss-sheet in the other two proteins. Traits 15-21 and 84-86 were exposed, while trait 91-93 was quite hidden in all the proteins. The antifungal activities of the four proteins towards the specific pathogenic fungus Fusarium culmorum were distinct and well correlated to the structural differences. These results suggest that these regions may have a role in the action mechanism, which is still unknown. PMID- 12638008 TI - Dynamical clustering of red blood cells in capillary vessels. AB - We have modeled the dynamics of a 3-D system consisting of red blood cells (RBCs), plasma and capillary walls using a discrete-particle approach. The blood cells and capillary walls are composed of a mesh of particles interacting with harmonic forces between nearest neighbors. We employ classical mechanics to mimic the elastic properties of RBCs with a biconcave disk composed of a mesh of spring like particles. The fluid particle method allows for modeling the plasma as a particle ensemble, where each particle represents a collective unit of fluid, which is defined by its mass, moment of inertia, translational and angular momenta. Realistic behavior of blood cells is modeled by considering RBCs and plasma flowing through capillaries of various shapes. Three types of vessels are employed: a pipe with a choking point, a curved vessel and bifurcating capillaries. There is a strong tendency to produce RBC clusters in capillaries. The choking points and other irregularities in geometry influence both the flow and RBC shapes, considerably increasing the clotting effect. We also discuss other clotting factors coming from the physical properties of blood, such as the viscosity of the plasma and the elasticity of the RBCs. Modeling has been carried out with adequate resolution by using 1 to 10 million particles. Discrete particle simulations open a new pathway for modeling the dynamics of complex, viscoelastic fluids at the microscale, where both liquid and solid phases are treated with discrete particles. Figure A snapshot from fluid particle simulation of RBCs flowing along a curved capillary. The red color corresponds to the highest velocity. We can observe aggregation of RBCs at places with the most stagnant plasma flow. PMID- 12638009 TI - Structures and magnetic properties of mono-doped fullerenes C59Xn and C59X(6mn)m (X=Bm, N+, P+, As+, Si): isoelectronic analogues of C60 and C60(6m). AB - Structures of mono-doped fullerenes, C59Xn and C59X(6mn)m (X=Bm, N+, P+, As+, Si), the isoelectronic analogues to C60 and C606m with 60 and 66 pi-electrons, have been investigated at the B3LYP/6-31G* level of density functional theory. On the basis of the computed nucleus independent chemical shifts (NICS) at the cage center and also at the center of individual rings as magnetic criteria, heterofullerenes with 60 pi-electrons are as aromatic as the parent C60, while those with 66 pi-electrons are much less aromatic than C606m. The very distinct endohedral chemical shifts of the 66 pi-electron systems may be useful to identify the heterofullerenes through their endohedral 3He NMR chemical shifts. PMID- 12638010 TI - Structure analysis of montmorillonite intercalated with rhodamine B: modeling and experiment. AB - The intercalation process and the structure of montmorillonite intercalated with [rhodamine B]+ cations have been investigated using molecular modeling (molecular mechanics and molecular dynamics simulations), X-ray powder diffraction and IR spectroscopy. The structure of the intercalate depends strongly on the concentration of rhodamine B in the intercalation solution. The presence of two phases in the intercalated structure was revealed by modeling and X-ray powder diffraction: (i) phase with basal spacing 18 A and with bilayer arrangement of guests and (ii) phase with average basal spacing 23 A and with monolayer arrangement of guests. In both phases the monomeric and dimeric arrangement can coexist in the interlayer space. Three types of dimers in the interlayer structure have been found by modeling: (i) H-dimer (head-to-head arrangement) present in the 18 A phase, (ii) sandwich type of the head-to-tail arrangement (present in the 23 A phase) and (iii) J-dimer (head-to-tail arrangement) present in the 23 A phase. Figure Montmorillonite intercalated with rhodamine B cations. On the left: phase 18 A, bilayer dimeric arrangement of guests (H-dimers). On the right: phase 23 A, monolayer arrangement of guests prepared using intercalation solution with a low concentration of rhodamine B PMID- 12638011 TI - Binding site characteristics in structure-based virtual screening: evaluation of current docking tools. AB - Two new docking programs FRED (OpenEye Scientific Software) and Glide (Schrodinger, Inc.) in combination with various scoring functions implemented in these programs have been evaluated against a variety of seven protein targets (cyclooxygenase-2, estrogen receptor, p38 MAP kinase, gyrase B, thrombin, gelatinase A, neuraminidase) in order to assess their accuracy in virtual screening. Sets of known inhibitors were added to and ranked relative to a random library of drug-like compounds. Performance was compared in terms of enrichment factors and CPU time consumption. Results and specific features of the two new tools are discussed and compared to previously published results using FlexX (Tripos, Inc.) as a docking engine. In addition, general criteria for the selection of docking algorithms and scoring functions based on binding-site characteristics of specific protein targets are proposed. Figure Enrichment factors obtained with FlexX, Glide and FRED docking engines in combination with different scoring functions for seven selected targets with highly variable binding sites PMID- 12638012 TI - Binding mode of ecdysone agonists to the receptor: comparative modeling and docking studies. AB - Three-dimensional structure models of the ligand-binding domain of the ecdysone receptor of Heliothis virescens were built by the homology modeling technique from the crystal structures of nuclear receptors. Two models were created based both on known ligand-binding domain structures of the receptors with the highest sequence identity to the ecdysone receptor, and on those of steroid hormone receptors. The latter model, which was found to have better stereochemical quality and be in good agreement with the binding of the steroidal framework of the endogenous agonist 20-hydroxyecdysone, was used for docking studies. The docking of 20-hydroxyecdysone to the receptor model revealed that the ligand molecule can interact with the receptor in a similar manner to other steroid hormone-receptor complexes. The docking of a dibenzoylhydrazine agonist, chromafenozide, was performed based on the correspondences between the molecule and 20-dydroxyecdysone expected by molecular comparison. The interactions of the ligands with the receptor in the complexes modeled were investigated and found to be consistent with known structure-activity relationships. PMID- 12638013 TI - The Compressed Feature Matrix--a novel descriptor for adaptive similarity search. AB - The Compressed Feature Matrix (CFM) is a new molecular descriptor for adaptive similarity searching. Depending on the requirements, it is based on a distance or geometry matrix. Thus, the CFM permits topological and three-dimensional comparisons of molecules. In contrast to the common distance matrix, the CFM is based on features instead of atoms. Each kind of these features may be weighted separately, depending on its (estimated) contribution to the biological effect of the molecule. In this work, we show that the CFM allows us to adapt similarity evaluations to particular ligands as well as to classification requirements. The CFM method is analyzed regarding correctness, adaptivity and speed. Applying the basic setting of feature weights, the similarity evaluations using the CFM on the one hand and the Tanimoto coefficient together with MACCS Keys on the other yield similar results. However, in contrast to the latter method, the CFM even permits us to focus on small parts of molecules to serve as a basis for similarity. Accordingly, we have achieved striking results not only by readjusting the feature weights with regard to the scaffold but also to the side chain of the respective target. The results of the latter run turned out to be rather independent of the molecular scaffold. Hence, the CFM is suitable not only for common similarity evaluation, but also for techniques such as lead or scaffold hopping. Figure Chemical structure, feature graph and topological CFM of serotonine PMID- 12638015 TI - Mitochondrial cytopathies. AB - Mitochondrial cytopathies represent a heterogeneous group of multisystem disorders which preferentially affect the muscle and nervous systems. They are caused either by mutations in the maternally inherited mitochondrial genome, or by nuclear DNA-mutations. Today, approximately 200 different disease causing mutations of mitochondrial DNA (mtDNA) are known, and due to the increased knowledge about nuclear genetics during the last few years, more and more nuclear mutations are being described. Owing to the non-uniform distribution of mitochondria in tissues and the co-existence of mutated and wildtype mtDNA (heteroplasmy) in these organelles, these disorders may present with a huge variety of symptoms, even if the same mutation is involved. Diagnostic investigations should include the measurement of serum and CSF lactate, neuroradiological tests and a muscle biopsy to show the characteristic ragged-red fibres and cytochrome c oxidase deficient cells and also to provide material for genetic analysis. To date, the treatment of these diseases remains supportive and should focus on typical complications such as cardiac dysrhythmia and endocrinopathy. PMID- 12638016 TI - Spectrum of the mitochondrial DNA mutations of Leber's hereditary optic neuropathy in Koreans. AB - We investigated 14 primary mitochondrial DNA (mtDNA) mutations at nucleotide positions (nps) 3460A, 4160C, 5244A, 9101C, 9804A, 10663C, 11778A, 13730A, 14459A, 14482G, 14484C, 14495G, 14498T, and 14568T, and one common secondary mutation at np 15257A, in 82 Korean patients with suspected Leber's hereditary optic neuropathy (LHON). Only three kinds of LHON mutations were identified in 60 (73 %) of the 82 probands, these being the 11778A, 14484C, and 3460A mutations with 46 (56%), 13 (16%), and 1 (1%) cases, respectively. None of the other mtDNA mutations was detected. Of the 60 probands with LHON positive mutations, 19 (32%) had relevant family histories. Heteroplasmy was determined in 2 (4%) of the 46 probands with the 11778A mutation and 1 (8%) of the 13 probands with the 14484C mutation. In conclusion, the 11778A mutation was the most common cause (56%), with a high prevalence of the 14484C and a lower prevalence of the 3460A mutations being characteristic of Korean patients with LHON. The 3460A mutation especially showed a remarkable racial difference from that in Caucasians. With the exceptions of the 3460A, 11778A, and 14484C, the mutations screened may not be involved in the pathogenesis of LHON in Koreans and may not have a synergistic effect on its clinical expression. PMID- 12638017 TI - Does personality change as a result of fetal tissue transplantation in the brain? AB - This study examined the effects on personality of transplantation of fetal tissue into the brains of participants in a double-blind placebo control trial for the treatment of Parkinson's disease (PD). Thirty persons with PD (equal numbers of males and females) participated in a larger study investigating the efficacy of transplantation of fetal neural tissue versus placebo surgery. Participants were randomly assigned to receive either the fetal transplant or placebo surgery. The blind was lifted for all patients approximately 13 months after surgery, at which time individuals who had received the placebo surgery could choose to receive the transplant surgery. In this study 12 persons originally received the transplant and 18 received placebo surgery. The NEO Five-Factor Inventory (NEO-FFI), a commonly used measure of personality characteristics, was administered to participants at baseline, 12, and 24 months after surgery. Scores at baseline for the Openness and Agreeableness scales were significantly higher for this sample of PD patients than scores for the normative group. There were no changes on any of the five scales from baseline to 12 months for the total group. The only significant change in the original transplant group was a decrease in Conscientiousness from baseline to 24 months. There were no changes over time among the group who had placebo surgery first and then the transplant. Results indicate that personality, as measured by the NEO-FFI, basically remained stable during the two-year follow-up period of this study. In this case, no change is regarded as a positive outcome. PMID- 12638018 TI - A study of the mechanisms of normal-appearing white matter damage in multiple sclerosis using diffusion tensor imaging--evidence of Wallerian degeneration. AB - Diffusion tensor imaging (DTI) investigates brain tissue microstructure in vivo. In multiple sclerosis (MS) Wallerian degeneration of axons traversing focal lesions is a potential mechanism of damage in normal-appearing white matter. In vivo evidence for this hypothesis is limited. The present study investigated the relationship between DTI-derived indices in the normal-appearing corpus callosum (CC) and the lesion loads (LLs) in connected cerebral regions. DTI was performed in 39 MS patients and in 21 age-matched controls. Fractional anisotropy (FA) and mean diffusivity (MD) were estimated in the genu, body and splenium of CC. Patients showed lower FA and higher MD in the CC than controls and both correlated with the total LL (r = -0.56 and r = 0.54, p < 0.0001). The LL of individual cerebral lobes correlated with both FA and MD in the corresponding callosal regions, with the body showing the strongest correlations with frontal and parietal LL (p < 0.0001). The strong correlations between DTI indices in the CC and the extent of lesions in connected brain regions support the hypothesis that Wallerian degeneration of axons transected by remote, but connected focal lesions, is an important pathogenic mechanism of damage in MS. PMID- 12638019 TI - Cycle ergometry is not a sensitive diagnostic test for mitochondrial myopathy. AB - Cycle exercise has repeatedly been used to diagnose patients suspected of having mitochondrial myopathy (MM), in whom exercise intolerance and lactic acidosis are common. No standardized test, however, has been established. We evaluated the diagnostic value of incremental and constant workload (20 min at 65 % VO(2max)) cycle tests for the diagnosis of MM. Plasma lactate and oxidative capacity (VO(2) and workload) were measured in 15 well-characterized MM patients during cycling. Findings were compared with those in 10 myotonic dystrophy patients and 18 sedentary, healthy subjects. All MM patients had ragged red or COX-negative fibers on muscle biopsy. VO(2max) and maximal workload were lower in MM than in control groups (P < 0.02). Resting plasma lactate was higher in MM than in control groups (P < 0.005; sensitivity = 93 %; specificity = 85 %), while exercise-induced increases in plasma lactate were only higher during the constant workload protocol in MM patients vs. control groups (P < 0.05; sensitivity = 27 %; specificity = 86 %). The findings indicate that the diagnostic value of a constant workload protocol is superior to an incremental cycle test, but that the test is less sensitive for MM than simple testing of resting lactate and muscle morphology. Cycle testing of MM patients remains an important research tool, but should not be a standard diagnostic procedure for MM. PMID- 12638020 TI - Cerebral proton magnetic resonance spectroscopy in infantile Alexander disease. AB - Alexander disease (AD) is a rare genetic disorder of the central nervous system due to a dysfunction of astrocytes. The most common infantile form presents as a progressive leukodystrophy with macrocephalus. Recently, heterozygous de novo mutations in the gene encoding glial fibrillary acidic protein (GFAP) have been demonstrated to be associated with AD. We used localized proton magnetic resonance spectroscopy (MRS) to assess metabolic abnormalities in grey and white matter, basal ganglia, and cerebellum of 4 patients with infantile AD and GFAP mutations. Strongly elevated concentrations of myo-inositol in conjunction with normal or increased choline-containing compounds in all regions investigated point to astrocytosis and demyelination. Neuroaxonal degeneration, as reflected by a reduction of N-acetylaspartate, was most pronounced in cerebral and cerebellar white matter. The accumulation of lactate in affected white matter is in line with infiltrating macrophages. Metabolic alterations demonstrated by in vivo proton MRS are in excellent agreement with known neuropathological features of AD. PMID- 12638021 TI - Magnetic resonance imaging of the cervical spinal cord in multiple sclerosis--a quantitative T1 relaxation time mapping approach. AB - BACKGROUND: The spinal cord is a common site of involvement in multiple sclerosis (MS) where pathology contributes substantially to locomotor disability. Previous studies have demonstrated significant correlations between clinical disability and cervical cord atrophy, but not with cord T(2) lesion load. We evaluate cervical cord pathology using, for the first time, quantitative T(1) relaxation time (T(1)), which shows histopathological specificity for tissue damage in the cerebral white matter. METHOD: Cervical cord T(1) was compared in 15 MS patients [8 relapsing-remitting (RR), 7 secondary progressive (SP)] and 6 healthy controls, and related to normalised upper cervical cord area (UCCa), cerebral white matter T(1), T(2) lesion load and disability measures including the Expanded Disability Status Scale (EDSS), Ambulation index (AI) and timed 25-foot walk. T1 maps of the brain and cervical cord were acquired using a high resolution, 3-dimensional fast low-angle shot sequence. Dual-echo sequences were also obtained. RESULTS: Median cervical cord T(1) [mean (standard deviation)] was significantly greater in RR [854 [28] ms] (p = 0.0006) and SP patients [927 [67] ms] (p < 0.0001) compared with controls [888 [61] ms], and in SP vs. RR patients (p = 0.002). In the overall patient cohort, it correlated significantly with median cerebral white matter T1 (r = 0.7, p = 0.0046), UCCa (r = -0.87, p < 0.0001), but not T2 lesion loads. Both median cervical cord T1 and UCCa (respectively) correlated significantly with the EDSS (r = 0.55, p = 0.03; r = 0.54, p = 0.04), AI (r = 0.77, p = 0.001; r = -0.60, p = 0.02) and timed 25-foot walk (r = 0.56, p = 0.03; r = -0.55, p = 0.04). CONCLUSION: Cervical cord T(1) distinguishes between MS subgroups and could also prove a useful surrogate outcome measure in MS. The relation of cervical cord T(1) to cerebral white matter T(1) suggests that cord pathology may be influenced by tissue damage upstream. PMID- 12638022 TI - A case of incomplete postganglionic cholinergic dysautonomia. AB - Postganglionic cholinergic dysautonomia is a relatively uncommon disease that manifests as an acute and selective dysfunction of the parasympathetic and sympathetic sudomotor nervous system. Here, we present a 27-year-old woman who suffered from subacute onset of bladder paresis. The clinical and laboratory examinations revealed a selective parasympathetic dysfunction including dilated pupils and decreased tearing without an involvement of the sympathetic sudomotor system. From clinical and laboratory evidences, it is suggested that the tonic pupil and postganglionic cholinergic dysautonomia may not be a completely different disease entity but share the same pathogenesis, which is separated only by the severity or the extent of the disease. The patient was diagnosed as 'incomplete postganglionic cholinergic dysautonomia'. PMID- 12638023 TI - R1--systemic thrombolysis in German stroke units--the experience from the German Stroke data bank. AB - BACKGROUND: Systemic thrombolysis with tissue plasminogen activator (t-PA) for treatment of acute ischemic stroke was approved in Germany in 2000. Up to now, only data from single centers have been available for the study of the use of thrombolysis in a hospital-based approach outside controlled trials. We therefore sought to determine the frequency of application and complications as well as the patient outcome after t-PA treatment in clinical routine of specialized stroke centers in Germany. METHODS: Within the German Stroke Data Bank Collaboration, 6234 consecutive patients with ischemic stroke were prospectively documented in 20 stroke centers between 1998 and 1999. The patients were centrally followed via telephone interview after 3 months and 1 year to assess global functional outcome using the Modified Rankin Scale. RESULTS: 250 patients (4 %) received systemic t PA treatment during the study period. The baseline characteristics of these patients were comparable to large clinical trials and phase IV studies. Symptomatic and asymptomatic parenchymal hemorrhage occurred in 22 patients (8.8 %) and was fatal in 3 patients. Follow-up data after 3 months were obtained in 82.4 % of all patients, of which 35 % had a favorable functional outcome (mRS /= 4) and 17 % had died. CONCLUSION: The results of our study agree with the assumption that thrombolytic therapy can be performed safely and effectively in daily clinical practice. Nevertheless, the small proportion of patients receiving thrombolysis even in specialized stroke centers calls for further improvement of acute stroke management in Germany. PMID- 12638024 TI - Cognitive impairment in chronic obstructive pulmonary disease--a neuropsychological and spect study. AB - Some analogy exists between cognitive impairment in hypoxemic patients with chronic obstructive pulmonary disease (COPD) and Alzheimer's disease (AD). We purposed to verify whether the analogy extends to the cerebral perfusion pattern. Ten normal subjects, 15 COPD patients with and 18 without hypoxemia, and 15 patients with mild AD matched for age and educational level underwent brain perfusion single photon emission computed tomography (SPECT) and neuropsychological assessment. Normal subjects and non hypoxemic COPD patients had comparable perfusion patterns. The average perfusion decreased from non hypoxemic to hypoxemic COPD and, then, to AD patients. Hypoperfusion of associative areas was the hallmark of AD, whereas the average perfusion of anterior cortical and subcortical regions did not distinguish AD and hypoxemic COPD patients. Both COPD groups scored higher than AD patients (p 15, the Beck Depression Inventory (BDI) and the Clinical Global Impression (CGI) Scale) were randomized to receive either 20 mg/d fluoxetine or placebo for 3 months. Beside the psychiatric assessment, patients were evaluated by use of the Scandinavian Stroke Scale (SSS), the Mini-Mental-State-Examination (MMSE) and the Barthel-Index (BI). An open-label long-term follow up was done 18 months after the initial assessment. RESULTS: 54 depressed patients of an inpatient population of 242 consecutive stroke patients aged 25 to 85 years entered the trial within the first two weeks post-stroke. 50 patients completed the trial per-protocol. The initial severity of depression was comparable in the two groups (mean baseline HDS score 32.8 in the fluoxetine vs. 30.3 in the placebo group), as were neurological symptom severity and demographic parameters. Significant improvement was seen in both groups within 4 weeks of treatment, whereas no advantages of fluoxetine could be observed at this time. This indicates a high degree of spontaneous recovery during early rehabilitation therapy. BDI scores of patients treated with fluoxetine further decreased until the follow-up at 12 weeks, whereas the scores increased again in the placebo group. This depressive relapse of the placebo patients after the end of most rehabilitation efforts was evident at a long-term follow-up 18 months after inclusion, when patients who had been treated with fluoxetine were significantly less depressed. No side effects of fluoxetine treatment were detected. CONCLUSIONS: The advantages of fluoxetine were obvious at the follow-up 18 months after inclusion, but could not be demonstrated within the first three months of controlled treatment. The multitude of therapeutic efforts that take place in the early phase of rehabilitation might have facilitated spontaneous recovery from depression and might have hindered benefits of antidepressant treatment to become obvious. Fluoxetine treatment was well tolerated and safe. PMID- 12638028 TI - Endocrine responses to nocturnal eating--possible implications for night work. AB - BACKGROUND: Night work is becoming more common and shift workers display several metabolic disturbances. Aim To study the endocrine responses in relation to time of day during a 24-h period and how dietary macronutrient composition affects these responses. DESIGN: Seven males (26-43 y and 19.9-26.6 kg. m(-2)) were studied in a crossover design. Isocaloric diets described as high-carbohydrates (HC; 65 energy percent (E%) carbohydrates and 20E% fat) or high-fat (HF; 40E% carbohydrates and 45E% fat) were given. After a 6-day diet adjustment period, the subjects were kept awake for 24 h in a metabolic unit and were served an isocaloric meal (continuation of respective diet) every 4-h. Blood samples were taken throughout the 24-h period. RESULTS: Insulin and leptin responses to meal intake differed with respect to time of day (p < 0.05). Time of day affected glucagon, thyroid stimulating hormone (TSH), free thyroxin (fT4), total triiodothyronine (tT3), cortisol, chromogranin A (CgA) and pancreatic polypeptide (PP) concentrations (p < 0.05). Meal intake decreased cortisol concentration after meals at 0800, 1200 and 0400 but not at 1600, 2000 and 0000 h. The PP's postprandial increase was greater during 0800-1600 h compared to 2000-0800 h. With the HC meals, lower glucagon and CgA concentrations (p < 0.05), and a tendency for lower tT3 concentrations (p = 0.053) were observed compared to the HF meals. CONCLUSION: Insulin, PP, TSH, fT4, cortisol and leptin responses to meal intake differed with respect to time of day. The decreased evening/nocturnal responses of cortisol and PP to meal intake indicate that nocturnal eating and night work might have health implications. PMID- 12638029 TI - The status of plasma homocysteine and related B-vitamins in healthy young vegetarians and nonvegetarians. AB - BACKGROUND: Exclusion of animal products and having only plant protein in vegetarian diets may affect the status of certain B-vitamins, and further cause the elevation of plasma homocysteine concentration. AIM: The purpose of this study was to assess the status of homocysteine and related B-vitamins in vegetarians and nonvegetarians. The effects of biochemical parameters of B vitamins and dietary protein on plasma homocysteine were also examined. METHODS: The study was performed at the Chung Shan Medical University, Taichung, in the central part of Taiwan. Thirty-seven vegetarians (28.9 +/- 5.5 y) and 32 nonvegetarians (22.9 +/- 1.6 y) were recruited. Nutrient intake was recorded using 3-day dietary records. Fasting venous blood samples were obtained. Plasma homocysteine, folate and vitamin B-12 were measured. Vitamin B-6 status was assessed by direct measures [plasma pyridoxal 5'-phosphate (PLP) and urinary 4 pyridoxic acid (4-PA)] and indirect measures [erythrocyte alanine (EALT-AC) and aspartate (EAST-AC) aminotransaminase activity coefficient]. RESULTS: There was no significant difference in vitamin B-6 intake between the two groups, although the vegetarian group had a significantly lower vitamin B-12 intake than the nonvegetarian group. Vegetarian subjects had significantly lower mean plasma PLP and vitamin B-12 concentrations than did nonvegetarian subjects (p < 0.05); however, a significantly higher mean plasma folate concentration was found in the vegetarian group. Vegetarian subjects had a significantly higher mean plasma homocysteine concentration than nonvegetarian subjects (13.2 +/- 7.9 vs. 9.8 +/- 2.2 micromol/L). Negative correlations were seen between plasma homocysteine and vitamin B-12 concentrations in the vegetarian (p = 0.004), nonvegetarian (p = 0.026), and pooled (p < 0.001) groups. From best subsets regression analyses, the plasma homocysteine concentration could be significantly predicted by total protein intake (p = 0.027) and plasma vitamin B-12 concentration (p = 0.005) in the pooled group. When the intake of protein is not considered, vitamin B-12 concentration is still a strong predictor of plasma homocysteine concentration (p = 0.012). CONCLUSIONS: Vitamin B-12 intake and mean plasma vitamin B-12 concentration were lower for vegetarian subjects than for nonvegetarian subjects, leading to an increase in plasma homocysteine concentration. Vitamin B-6 and folate had little effect on plasma homocysteine concentration when individuals had adequate vitamin B-6 and folate status. PMID- 12638030 TI - Effects of inulin-type fructans of different chain length and type of branching on intestinal absorption and balance of calcium and magnesium in rats. AB - BACKGROUND: Inulin-type fructans or chains with mainly beta[2-1] linked fructose molecules escape the ingestion procedure in the small intestine and are fermented by the microflora, and are known to increase colonic absorption of minerals in animals. The fermentation rate in the large bowel into short-chain fatty acids depends on the molecular mass and the structure of these food ingredients. It is thought that this colonic fermentation is the basis for the reported increase in mineral absorption. AIM OF THE STUDY: The purpose of the present study was twofold: a) to compare different types of fructans that differ in the sugar chain length and in chain branching; b) to determine the potential synergistic effect of a combination of inulin-type fructans with different chain lengths. METHODS: For this purpose, 50 adult male Wistar rats weighing 170 g each were used in this study. The rats were distributed into 5 groups and fed for 28 days a fiber-free basal purified diet or diet containing 10 % oligofructose (OF) (DP(av) 4), or 10 % HP-inulin (DP(av) 25), a blend of 50 % OF and 50 % HP-inulin, or a branched chain inulin. RESULTS: During the first period, the rats went into a gradual adaptation, during which the rats received 2.5 % for 1 week and then 5 % for 1 week of the tested products. During the last 4 days of the experiment, feces and urine were monitored for mineral balance study. The animals were then sacrificed and blood, cecum and tibia were sampled for mineral status assessment. Our results showed that the ingestion of all the tested fructans led to a considerable cecal fermentation. All tested compounds increased the intestinal absorption and balance of Mg significantly. Interestingly, in the present experimental set-up, all tested compounds increased the intestinal absorption and balance of Ca numerically, but only the blend OF + HP-inulin increased apparent intestinal absorption and balance of Ca significantly. CONCLUSIONS: The different types of fructans studied in the present experiment seem to have similar activity on mineral absorption. However, the combination of OF and HP-inulin showed synergistic effects on intestinal Ca absorption and balance in rats. Further studies with other combinations of fructans need to be done to extend these findings. PMID- 12638031 TI - Nutritional concerns and possible countermeasures to nutritional issues related to space flight. AB - The experience obtained in astronauts until now has revealed many nutritional problems during both short- and long-term missions that are still under investigation. When the manned space program evolves to the point where it involves a considerable number of people operating at great distances from the Earth for long periods of time, continuous provision of food as well as physiopathological modifications partly related to nutrition will remain a challenge. Human space missions have outlined the importance of the diet on the quality of an astronaut's life, not only because an appropriate nutritional status can be maintained only through an adequate nutrient intake, but also because food plays an important socio-psychological role.A great amount of research has been done both by the Russian and American teams in order to identify the nutritional requirements for humans during space flights. Crew members should be provided with nutritional adequate diets characterized by many different food items carefully selected according to technological techniques aimed to ensure long shelf-life periods, health, safety, satisfaction and convenience. An astronaut's nutritional status is greatly influenced by important weightlessness, environmental physiological adaptations.Changes in muscle and bone mass, modifications of gastro-intestinal functions and immune alterations may be partly limited by adequate planned dietary countermeasures. PMID- 12638032 TI - A phytosterol-enriched spread improves the lipid profile of subjects with type 2 diabetes mellitus--a randomized controlled trial under free-living conditions. AB - BACKGROUND: Phytosterol-enriched margarines are known to significantly lower total and LDL cholesterol, but little is known about the effect of such margarines in subjects with type 2 diabetes. AIM OF THE STUDY: Investigation of the effect of a phytosterol-enriched spread in subjects with type 2 diabetes mellitus on serum lipids, Hb(A1c), and blood glucose under free-living conditions. METHODS: Randomized, placebo-controlled, double-blind clinical trial in two parallel groups over 12 weeks; 85 type 2 diabetic patients with serum LDL cholesterol levels >/= 3.60 mmol/l and without hypolipidemic medication were included in the study. Participants consumed 2 x 10 g of spread with or without 8 % phytosterol-esters daily. Fasting blood samples were analyzed at 0, 4, 8, and 12 weeks. RESULTS: After 4 weeks, total and LDL cholesterol were significantly reduced in the phytosterol group by 5.2 % and 6.8 %, respectively, compared to baseline (p < 0.05). After 8 and 12 weeks, these reductions became smaller and were not significant any more compared to baseline or between the groups, but a repeated measurement analysis demonstrated a significant difference for both variables between the two groups (each p < 0.05). HDL cholesterol was significantly increased in the phytosterol group compared to the placebo group after 8 and 12 weeks, but there was no overall difference in the repeated measurement analysis between the two groups. In the phytosterol group, there was a small reduction in Hb(A1c) compared to the control group which was only significant after 4 weeks. CONCLUSIONS: This clinical study shows that a phytosterol-enriched spread is effective in lowering total and LDL cholesterol in subjects with type 2 diabetes but also illustrates the difficult maintenance under free-living conditions over time. Although this effect is modest, it may contribute to decreasing the elevated risk of cardiovascular disease in type 2 diabetes. PMID- 12638033 TI - Vitamins and minerals: a model for safe addition to foods. AB - BACKGROUND: Significant subgroups in most European populations have intakes below nationally recommended levels for several vitamins, minerals and trace elements, placing individuals at risk of suboptimal intake of important vitamins and minerals. The voluntary addition of micronutrients to the appropriate foods may help address the risks associated with low micronutrient intakes. However, concerns need to be addressed regarding the potential for unacceptably high intakes, particularly for those people consuming very large amounts of food. AIM OF THE STUDY: To develop a model to estimate the level of each micronutrient that can be added safely to foods. METHODS: A theoretical model was developed based on the critical factors which determine the risk of unacceptably high intake for each micronutrient at high levels of food/energy intakes. These included 1) Tolerable Upper Intake Levels (UL), 2) high micronutrient intakes in Europe at the 95(th) percentile intake for each nutrient, 3) the proportion of fortified foods in the diets of individuals at the 95(th) percentile for energy intakes, 4) the proportion of foods to which micronutrients could practically be added, and 5) a range of estimates for the fractions of foods which might be actually fortified for each nutrient. A maximum level was set up for each micronutrient per typical serving or 100 kcal portion. The outputs of the model were then compared against a recent model developed by AFSSA, based on the food intake data in France. RESULTS: Three categories of micronutrients were identified, in which micronutrients could be added safely to foods at levels (per serving, e. g., 100 kcal) 1) greater than 1 European Commission Recommended Daily Intake (EC RDA): vitamin B12, vitamin C, vitamin E, riboflavin, panthothenic acid, niacin and thiamine; 2) between 50 and 100 % of the EC RDA: vitamin B6, vitamin D, folic acid, biotin, copper, iodine and selenium; 3) between 10 and 40 % of the EC RDA: iron, zinc, calcium, phosphorus and magnesium. A fourth category consisting of retinol, for which high end intake levels are close to UL for some population subgroups in Europe and thus requires further consideration. CONCLUSIONS: A wide range of vitamins and minerals can be added safely to foods at nutritionally important levels in the current diets of Europeans. PMID- 12638034 TI - A benign colorectal stricture: treatment with a retrievable expandable nitinol stent. PMID- 12638035 TI - Combined transabdominal and transjugular recanalization of thrombosed TIPS, portal and proximal splenic veins. AB - We present a case of TIPS occlusion in which a small caliber transhepatic safety or anchoring wire in combination with a standard transjugular approach were utilized to recanalize the TIPS, portal and splenic veins. This technique may be a useful adjunct to the typical methods used for recanalization of thrombosed TIPS, portal and splenic veins without the need of large caliber transhepatic sheath access. PMID- 12638036 TI - Supplemental transcatheter arterial chemoembolization through a collateral omental artery: treatment for hepatocellular carcinoma. AB - PURPOSE: To evaluate the therapeutic efficacy and safety of supplemental transcatheter arterial chemoembolization (TACE) through the extrahepatic collateral omental artery (OA) for the treatment of hepatocellular carcinoma (HCC). METHODS: We studied 21 patients with extrahepatic collaterals of the OA, among 1,512 patients with HCC who had undergone angiography. HCCs supplied by collateral OAs were located at: segment IV in seven, segment V in five, segment III in three, segment VI in three and segment VIII in three patients (Couinaud classification of segments). On preoperative CT scans, every HCC was abutting the liver surface. Adjacent omental infiltration or engorgement was noted in 11 patients. Celiac and hepatic arteriograms showed hypertrophy of the feeding OA in all patients. TACE of the OA was performed in 19 patients with an emulsion of iodized oil and doxorubicin hydrochloride. Embolization with gelatin sponge particles was added in five patients. RESULTS: Collaterals of the OA to the HCC were found on the first to seventeenth sessions of TACE. On follow-up CT scans, five patients showed complete uptake of iodized oil in the tumor. Partial uptake of iodized oil was noted in 13 patients and no uptake in one patient. There was no serious complication that related to the omental embolization, such as omental or bowel ischemia. The cumulative survival rates from the time of the TACE of the OA were 81% at 6 months and 68% at 12 months. CONCLUSION: TACE of the OA is safe and has a potential therapeutic effect in the treatment of HCC. PMID- 12638037 TI - Common bile duct stones detected after cholecystectomy: advancement into the duodenum via the percutaneous route. AB - PURPOSE: To report our experience in the use of percutaneous extraction of common bile duct stones detected in the post-cholecystectomy period. METHODS: Forty-two patients in whom endoscopic cannulation and/or sphincterotomy had failed or could not be done due to several reasons underwent balloon dilatation of the ampulla of Vater and subsequent advancement of the stones via the percutaneous transhepatic route or T-tube tract. RESULTS: The procedure was successful in 42 cases. In three patients, stones were crushed in the common bile duct and pushed as fragments into the duodenum. In all cases transient adverse effects were observed. There were no major complications. All cases were checked with ultrasonography for 6 months after the procedure. CONCLUSION: Percutaneous extraction of common bile duct stones is an effective method of treatment with a high success rate, low complication rate and shorter hospital stay. It may serve as an alternative method in cases where endoscopic removal of stones fails. PMID- 12638038 TI - Percutaneous interventions in radiation-associated coronary in-stent restenosis. AB - This study was performed to evaluate the outcome of percutaneous revascularization in "edge restenoses" developing after radioactive stent implantation in de novo and in-stent lesions. Twenty-one consecutive patients undergoing target lesion revascularization (TLR) at any follow-up after phosphorus-32 radioactive stent implantation were included in this study. We assessed the incidence of death, myocardial infarction, repeated TLR and recurrent angina over the following 18 months. After 6 months, TLR rate was 28.6%, and no stent thromboses, deaths or Q-wave myocardial infarctions occurred. Among the patients with TLR there were significantly more subjects who had received a radioactive stent in a previous in-stent restenosis (66.7% vs. 0% in patients without second restenosis; P <0.001), or who had received two radioactive stents (83.3% vs. 33.3%; P = 0.038). After 18 months, TLR rate was 33.3%, and two patients (9.5%) had died. Restenosis after intravascular radiotherapy can be safely treated by percutaneous interventional techniques, yielding an acceptable clinical result within 18 months. PMID- 12638041 TI - The sky is not falling...accepting failure to create innovation in experimentation. PMID- 12638042 TI - Structural characterization of neutral oligosaccharide mixtures through a combination of capillary electrochromatography and ion trap tandem mass spectrometry. AB - A CEC/ESI-MS/MS combined system has been developed for the separation and on-line structural analysis of neutral oligosaccharides. Various types of isomeric oligosaccharides were first successfully separated by CEC using polar monolithic columns, while the on-line tandem mass spectrometry has been explored to differentiate and elucidate the structures of isomeric oligosaccharides. The experimentally obtained tandem spectra usually provide sequence, branching, and linkage information. Oligosaccharide isomers with a different monomeric composition and branching showed different patterns of glycosidic linkage cleavage (B- and Y-ion series), allowing us to deduce their sequence and branching points. Isomers with different linkages were distinguished by identifying cross-ring fragment ions (A-ion series). While (1-->4) linkages yielded dominant (0,2)A ions, (1-->6) linkages showed an extensive and complete cross-ring cleavage series: (0,2)A, (0,3)A, and (0,4)A ions. Although the anomeric configurations and monosaccharide identification are rarely obtained from tandem MS, the relevant mixture components can be completely resolved with high-efficiency CEC columns featuring a polar functionality. PMID- 12638043 TI - Quantification of bile acids directly from urine by MALDI-TOF-MS. AB - The ability to quantify mixtures of bile acids using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry directly from urine has been demonstrated. Six cholic acid derivatives were selected for analysis: taurocholic acid (TCA), taurochenodeoxycholic acid (TCDCA), taurolithocholic acid (TLCA), glycocholic acid (GCA), glycochenodeoxycholic acid (GCDCA), and glycolithocholic acid (GCDCA). Urine samples were pre-concentrated and purified using solid-phase extraction (SPE) columns. The method was optimized to eliminate suppression effects, and proved to be reproducible from day to day. Calibration curves averaged from three days were obtained for the bile acids directly from urine, and then tested for their ability to accurately determine concentrations from one measurement. In summary, a simple, rapid method has been developed for the quantification of bile salts from urine with SPE clean-up by MALDI-MS. PMID- 12638044 TI - Determination of hydrocortisone, polymyxin B and Zn-bacitracin in pharmaceutical preparations by micellar electrokinetic chromatography. AB - A new, simple and accurate micellar electrokinetic chromatography (MEKC) method is established for quantification of hydrocortisone, polymyxin B and Zn bacitracin in local pharmaceutical preparations. The separation was carried out at 25 degrees C and 25 kV, using a 15 mmol L(-1) phosphate-15 mmol L(-1) borate buffer (pH 8.2), 60 mmol L(-1) sodium dodecylsulfate (SDS), and 10% methanol water (v/v) as background electrolyte. Under these conditions the analysis takes about 23 min. The method has been applied for quantifying these compounds in two different commercial pharmaceutical products and the method gave good results when compared with a reference spectrophotometric multivariate calibration method. PMID- 12638039 TI - Non-invasive in vivo imaging of myocardial apoptosis and necrosis. AB - Myocardial necrosis plays an important role in the pathogenesis of various cardiovascular disorders and can result from different myocardial insults. Its non-invasive identification and localisation therefore may help in the diagnosis of these disorders, as well as in prognosis and assessment of treatment response. Apoptosis, or programmed cell death, is important in the spectrum of myocardial damage since it is gradually becoming more apparent that cell death may begin as apoptosis and not as necrosis. First attempts to directly visualise the area of myocardial necrosis were based on recognition of myocardial infarction with "hot spot imaging agents" in patients with chest pain. Since then, the study of myocardial necrosis with gamma imaging agents has gone beyond the detection of myocardial infarction, and attempts have been made to diagnose other cardiovascular disorders associated with cardiac cell death such as heart transplant rejection, myocarditis, cardiotoxicity and cardiomyopathies. Traditionally, two hot spot imaging agents have been used for the detection of myocardial necrosis, (99m)Tc-pyrophosphate and (111)In-antimyosin. In addition, preliminary studies have demonstrated promising results with (99m)Tc-glucarate. Recently, (99m)Tc-annexin V has been successfully used for non-invasive gamma imaging of apoptosis after acute myocardial infarction, acute myocardial ischaemia, acute cardiac allograft rejection and malignant intracardiac tumours. This review article focusses on the characteristics of these different myocardial necrotic and apoptotic markers and compares their role in the assessment of myocardial damage. PMID- 12638045 TI - Direct determination of four fluoroquinolones, enoxacin, norfloxacin, ofloxacin, and ciprofloxacin, in pharmaceuticals and blood serum by HPLC. AB - A rapid, accurate and sensitive method has been developed for the quantitative determination of four fluoroquinolone antimicrobial agents, enoxacin, norfloxacin, ofloxacin and ciprofloxacin, with high in-vitro activity against a wide range of Gram-negative and Gram-positive organisms.A Kromasil 100 C(8) 250 mm x 4 mm, 5 microm analytical column was used with an eluting system consisting of a mixture of CH(3)CN-CH(3)OH-citric acid 0.4 mol L(-1) (7:15:78 %, v/v). Detection was performed with a variable wavelength UV-visible detector at 275 nm resulting in limits of detection: 0.02 ng per 20 microL injection for enoxacin and 0.01 ng for ofloxacin, norfloxacin and ciprofloxacin. Hydrochlorothiazide (HCT) was used as internal standard at a concentration of 2 ng microL(-1). A rectilinear relationship was observed up to 2 ng microL(-1) for enoxacin, 12 ng microL(-1) for ofloxacin, 3 ng microL(-1) for norfloxacin, and 5 ng microL(-1) for ciprofloxacin. Separation was achieved within 10 min. The statistical evaluation of the method was examined by performing intra-day (n=8) and inter-day precision assays (n=8) and was found to be satisfactory with high accuracy and precision. The method was applied to the direct determination of the four fluoroquinolones in human blood serum. Sample pretreatment involved only protein precipitation with acetonitrile. Recovery of analytes in spiked samples was 97+/ 6% over the range 0.1-0.5 ng microL(-1). PMID- 12638047 TI - Separation of positional isomers by the use of coupled shape-selective stationary phase columns. AB - The successful separation of 2- and 3-methyl-substituted positional isomers of butanol, butyl acetate, and butanoic acid and its ethyl ester, is reported. These compounds are of interest in the study of wine flavour, however the separation of the 2- and 3-methyl isomers may present problems, and more so in the presence of the wine matrix components, when single capillary column gas chromatography (GC) is used. The strategy to achieve separation was based on the use of shape selective cyclodextrin derivative (CDD) capillary columns (commonly referred to as chiral columns). These columns provide simultaneous resolution of the enantiomeric pairs of the 2-methyl isomers, and at the same time the ability to separate the 3-methyl isomer from the 2-methyl is achieved in all but the case of the ( S)-2- and 3-methylbutanol. The advantages of using shape-selective columns to perform this study is demonstrated, with coupling of two CDD columns giving improved separations of these compounds. Although these compounds are relatively volatile, cryogenic modulated comprehensive two-dimensional GC was shown to provide good pulsed peak profiles with chiral separation in the first dimension when a thicker film trapping column segment was employed. The components of interest were well separated from other wine matrix components. PMID- 12638046 TI - Solid-state 29Si MAS NMR studies of diatoms: structural characterization of biosilica deposits. AB - Four different diatom species (Chaetoceros debilis, Chaetoceros didymum, Cylindrotheca fusiformis, Nitzschia angularis) were studied by solid-state (29)Si MAS NMR spectroscopy. To determine the Q(2):Q(3):Q(4) ratios in the biosilica deposits of the diatoms, quantitative (29)Si MAS NMR experiments were performed. This analysis did not reveal any differences regarding the molecular architecture of the silica (i.e. the degree of condensation of the SiOH units (2 identical with SiOH --> identical with Si-O-Si identical with + H(2)O)) from the different diatom species. However, complete cells showed significantly smaller Q(4):Q(3) ratios (1.8-1.9) than extracted cell walls (2.5-2.8), indicating the existence of intracellular pools of less condensed silica. PMID- 12638048 TI - Characterisation of quaternary mixtures by the apparent content curves method: identification of tocopherols in vegetable oils. AB - A procedure for identification of the compounds in quaternary mixtures has been developed. The proposed procedure is based on the apparent content curves method. From these curves and using the Q parameter, work conditions are selected and quaternary mixtures treated as "pseudoternary" mixtures.A simple strategy to test matrix effects at working wavelengths has been developed, identification limits established and following the identification table quaternary mixtures were easily characterised. The procedure has been applied to the identification of phenols in mixtures by UV-visible spectrophotometry and tocopherols in edible oils by fluorimetry. Results obtained for edible oils are in agreement with the composition obtained by use of the reference method. PMID- 12638049 TI - A comparative study of the correction of systematic errors in the quantitation of pyrethroids in vegetables using calibration curves prepared using standards in pure solvent. AB - A comparative study of two mathematical approaches was performed in order to correct systematic errors due to the presence of the unexpected interferences which appear when the quantitation of the analyte in real samples is carried out with calibration curves built using standards in pure solvent. These methods consisted in the establishment of different mathematical expressions which transform the concentration (Cs) obtained using calibration graphs built using pure solvent into the corrected concentration (C(M)) that should be obtained if the quantitation is carried out with calibration curves built using standards dissolved in blank matrix extracts. In the two approaches the correction is performed from the results of an intermediate precision study which was carried out using both calibration graphs (prepared using pure solvent and blank matrix extract). By using ANCOVA to compare the slope of both solvent-based and matrix matched calibration graphs, matrix effect was found in the determination of deltamethrin in tomato and acrinathrin in tomato and pepper. In these cases, both approaches led to good results. PMID- 12638050 TI - Spectrofluorimetric determination of formaldehyde by a flow-injection method based on its catalytic effect on the acridine yellow-bromate reaction. AB - A flow-injection configuration for the determination of formaldehyde is proposed. The method is based on the enhancing effect of formaldehyde on the oxidation of acridine yellow by bromate in acidic medium. The proposed procedure is simple, inexpensive, sensitive and suitable for concentrations of formaldehyde between 1 and 56 microg mL(-1). A sampling-rate of 60 samples h(-1) was achieved. The effect of several organic and inorganic species was studied. The method was applied to the determination of formaldehyde in pharmaceuticals, milk and air in work environments. The accuracy of the method was confirmed by comparing the results with those obtained using the standard acetylacetone method. PMID- 12638051 TI - The effect of the presence of volatile organoselenium compounds on the determination of inorganic selenium by hydride generation. AB - As a result of microbiological activity it is possible to find dimethylselenium (DMSe) and dimethyldiselenium (DMDSe) in a wide type of environmental samples, such as soils, sediments, sewage sludges and plants where methylation can take place. Selenium determination by hydride-generation (HG) techniques requires its presence as Se(IV). Consequently, inorganic speciation by hydride generation techniques is done by first determining Se(IV) and then, after reduction of Se (VI) to Se(IV), the total selenium. Therefore, the concentration of Se (VI) is evaluated as the difference between total inorganic selenium and Se(IV). In the present work it could be demonstrated that DMSe and DMDSe are forming other volatile species by reaction with sodium borohydride, applying the same reduction condition as for inorganic selenium. These species are subsequently detected by several atomic techniques (atomic absorption AAS, atomic fluorescence AFS and inductively coupled plasma-mass spectrometry ICP-MS). The error that their presence can cause in determination of inorganic selenium has been evaluated. The magnitude of this error depends on the specific analytical detector used.The coupling of pervaporation-atomic fluorescence is proposed for the identification of these species and pervaporation-gas chromatography-atomic fluorescence for their individual quantification. PMID- 12638052 TI - Microwave sample-digestion procedure for determination of arsenic in moss samples using electrothermal atomic absorption spectrometry and inductively coupled plasma mass spectrometry. AB - Arsenic in moss samples was determined by electrothermal atomic absorption spectrometry (ETAAS) after microwave-assisted sample digestion. Two different sample masses (500 mg and 1000 mg) and three different microwave ovens were used in the digestion. There was a slight difference in the digestion efficiency, as determined by the residual carbon concentrations of 500 mg digested samples, between the microwave ovens. The arsenic results obtained for moss reference samples were, in most cases, satisfactory. However, phosphorus was found to have a reducing influence on the arsenic peak area in the ETAAS determination. According to the results, it was not possible to reduce the phosphorus interference by increasing the amount of Mg(NO(3))(2) in the Pd-Mg chemical modifier. The arsenic results obtained by ETAAS were compared to those obtained by inductively coupled plasma mass spectrometry (ICP-MS). PMID- 12638053 TI - Separation of arsenic species by capillary electrophoresis with sample-stacking techniques. AB - A simple capillary zone electrophoresis procedure was developed for the separation of arsenic species (AsO(2)(2-), AsO(4)(2-), and dimethylarsinic acid, DMA). Both counter-electroosmotic and co-electroosmotic (EOF) modes were investigated for the separation of arsenic species with direct UV detection at 185 nm using 20 mmol L(-1) sodium phosphate as the electrolyte. The separation selectivity mainly depends on the separation modes and electrolyte pH. Inorganic anions (Cl(-), NO(2)(-), NO(3)(-) and SO(4)(2-)) presented in real samples did not interfere with arsenic speciation in either separation mode. To improve the detection limits, sample-stacking techniques, including large-volume sample stacking (LVSS) and field-amplified sample injection (FASI), were investigated for the preconcentration of As species in co-CZE mode. Less than 1 micromol L(-1) of detection limits for As species were achieved using FASI. The proposed method was demonstrated for the separation and detection of As species in water. PMID- 12638055 TI - PVC-membrane ion-selective bulk optode for Ag+ ion based on hexathia-18-crown-6 and 1,2-benzo-3-octadecanoylimino-7-diethylaminophenoxazine. AB - A silver-selective optode membrane incorporating hexathia-18-crown-6 for cation recognition and a lipophilic chromoionophore 1,2-benzo-3-octadecanoylimino-7 diethylaminophenoxazine for transduction has been prepared. The PVC membrane composition was optimized to result in the largest working concentration range. The response range of the proposed optode is 5.0 x 10(-9)-5.0 x 10(-5) mol L(-1) Ag(+) with a limit of detection of 1.0 x 10(-9) mol L(-1). The probe works well at pH 5.0 and revealed small ion interference and good selectivity, reproducibility and stability. PMID- 12638054 TI - Solid-phase spectrophosphorimetric determination of the pesticide o-phenylphenol in water and vegetables. AB - A phosphorimetric method for the determination of o-phenylphenol (OPP) using filter paper as solid support and Tl(I) as heavy metal enhancer of the phosphorescent signal is proposed. The phosphorescence measurements were carried out by placing the paper with the sample between two plates of quartz, thus avoiding the quenching effect produced by atmospheric oxygen and moisture. The linear dynamic range of the method was 0.5-4.0 mg L(-1) and the detection and quantification limits were 0.03 and 0.11 mg L(-1), respectively. The precision of the method (expressed as relative standard deviation) was 1.7% for a sample containing 2.0 mg L(-1) of analyte. The method has been applied to the determination of OPP in different types of water, lettuce, string beans and peppers, with recoveries ranging between 97.1 and 100.7% PMID- 12638056 TI - Solid-phase extraction-spectrophotometric determination of uranium(VI) in natural waters. AB - A method for the extraction and determination of uranyl ion in natural waters using octadecyl bonded silica membrane disks modified with piroxicam and spectrophotometry with arsenazo(III) is proposed. The perconcentration step was studied with regard to experimental parameters such as amount of extractant, type and amount of eluent, pH, flow rates and tolerance limit of diverse ions on the recovery of uranyl ion. The limit of detection of the proposed method is 0.4 micro g L(-1) of uranyl. The method was applied to the recovery of uranyl from different water samples. PMID- 12638057 TI - Simultaneous determination of 2-nitrophenol and 4-nitrophenol based on the multi wall carbon nanotubes Nafion-modified electrode. AB - In this work, multi-wall carbon nanotubes (MWNT) were conveniently dispersed into Nafion-ethanol solution, and the MWNT-Nafion-modified glassy carbon electrode (GCE) was described for the simultaneous determination of 2-nitrophenol and 4 nitrophenol. At pH 4.0 phosphate buffer, the reduction peak currents of 2 nitrophenol (at -0.8 V) and 4-nitrophenol (at -1.0 V) increase significantly at the MWNT-Nafion-modified GCE, in comparison with that at the Nafion-modified GCE and the bare GCE. The experimental parameters, such as solution pH of phosphate buffer, accumulation potential and time, and the amounts of MWNT-Nafion onto the GCE surface, were optimized. The reduction peak currents are linear with the concentration of 2-nitrophenol from 5 x 10(-8) to 1 x 10(-5) mol L(-1) and with that of 4-nitrophenol from 1 x 10(-7) to 1 x 10(-5) mol L(-1). The detection limits after 3-min accumulation are 1 x 10(-8) mol L(-1) for 2-nitrophenol and for 4 x 10(-8) mol L(-1) for 4-nitrophenol. This modified electrode was applied to direct determination of 2-nitrophenol and 4-nitrophenol in lake water samples. PMID- 12638058 TI - Co2+-selective membrane electrode based on the Schiff Base NADS. AB - A new PVC membrane electrode for cobalt(II) ions based on a recently synthesized Schiff base of 5-((4-nitrophenyl)azo)- N-(2',4'-dimethoxyphenyl)salicylaldimine is reported. The electrode exhibits a Nernstian response for Co(2+) ions over a wide concentration range (9.0 x 10(-7)-1.0 x 10(-2) M) with a slope of 29(+/-1). The limit of detection is 8.0 x 10(-7) M. The proposed sensor revealed good selectivities over a wide variety of other cations including hard and soft metals. This electrode could be used in a pH range of 3.5-6.0. It was used as an indicator electrode in potentiometric titrations of cobalt(II) ions and can be used in the direct determination of Co(2+) in aqueous solutions. PMID- 12638059 TI - CD spectrometric methods for the simultaneous determination of ethisterone and its delta5-isomer. AB - Quick and accurate direct and indirect circular dichroism (CD) spectrometric methods were developed for the simultaneous determination of ethisterone (17alpha ethinyl-17-hydroxy-4-androstene-3-one) and its delta(5)-isomer (delta(5) ethisterone). The direct method is based on the selective negative Cotton effect of the delta(4)-3-oxo group in ethisterone (negative maximum at 348 nm in dioxan) and measurement of the ellipticity at 296 nm (positive maximum of delta(5) ethisterone), where the measured ellipticity is the sum of those of the two isomers. In the indirect procedure delta(5)-ethisterone is transformed to ethisterone by base-catalysed isomerization and the ellipticities are measured at 339 nm in ethanol before and after isomerization. Preliminary experiments show the usefulness of CD detector in the HPLC determination of the mixture of the isomers. A major advantage of the direct CD spectrometric and the HPLC/CD methods is that the delta(5)-isomer with extremely low UV activity can also be directly measured with high sensitivity. PMID- 12638060 TI - Executive function in children: introduction. PMID- 12638061 TI - Assessment and development of executive function (EF) during childhood. AB - This review paper outlines the issues associated with the assessment of executive function (EF) in children and adolescents, and describes the developmental profile of executive processes across childhood. At the outset, EF is defined, and cognitive and behavioral impairments associated with executive dysfunction (EDF) are described. A developmental model of EF is proposed incorporating four discrete but inter-related executive domains (attentional control, cognitive flexibility, goal setting, and information processing) which operate in an integrative manner to enable "executive control". Characteristics that constitute traditional EF measures are discussed, as are the problems associated with test interpretation. The ecological validity of EF tests and neuropsychological assessment procedures are examined, and adjunct methods of measurement are presented to enable a more comprehensive and valid assessment of EF. Based on developmental and normative studies, the maturation of executive domains is mapped. Attentional control appears to emerge in infancy and develop rapidly in early childhood. In contrast, cognitive flexibility, goal setting, and information processing experience a critical period of development between 7 and 9 years of age, and are relatively mature by 12 years of age. A transitional period is thought to occur at the beginning of adolescence, and shortly after "executive control" is likely to emerge. In order to confirm our current understanding of EF development and further enhance our understanding of brain behavior relationships, longitudinal studies incorporating structural and functional neuroimaging are required. PMID- 12638062 TI - Executive functions in preschool children born preterm: application of cognitive neuroscience paradigms. AB - Although children born preterm are at risk for neuropsychological impairments at school age and adolescence, including difficulties with visual motor integration, spatial/constructional skills, attention, arithmetic, and nonverbal executive functions, specific neuropsychological outcome has not been investigated adequately in preschoolers. Application of cognitive neuroscience tasks offers the opportunity to characterize early executive functions in young children born preterm. In a preliminary sample of 29 preschool children born preterm (M birth gestational age = 32.4 weeks), executive function outcome was compared to that of fullterm controls by contrasting performance on two prototypic delayed-response type paradigms, Delayed Alternation and Spatial Reversal. Preschoolers born preterm correctly retrieved the reward on fewer trials on Delayed Alternation than did matched controls. Furthermore, preschool children born preterm used problem-solving strategies that included more perseverative errors than controls. These preliminary findings highlight the utility of cognitive neuroscience paradigms to understand neuropsychological outcome in preschool children born preterm and suggest areas of developmental vulnerability that may include dorsolateral prefrontal circuits. PMID- 12638063 TI - Planning and problem solving skills following focal frontal brain lesions in childhood: analysis using the Tower of London. AB - Problem solving skills were investigated in children with focal lesions using the Tower of London test (TOL; Shallice, 1982). The scoring procedure was elaborated from previous studies to delineate separate processes contributing to overall performance in children. Thirty-one children with focal frontal pathology, 18 children with focal pathology in other brain regions (extra-frontal), 17 children with generalized pathology and 38 healthy children participated in the study. Results suggest a distributed network for problem solving skills, particularly cognitive flexibility and goal setting skills. Within the frontal group, children with lesions involving the right pre-frontal cortex had greatest problems with self-regulation, with rule breaks most common among this group. As these skills develop relatively early in comparison to other aspects of executive function, right pre-frontal regions may play a particularly important role in the development of executive skills in childhood, with damage to these regions rendering children vulnerable to a range of cognitive and social deficits. PMID- 12638064 TI - Selective learning in children after traumatic brain injury: a preliminary study. AB - Selective learning (SL) is the ability to select items to learn from among other items. It requires the use of the executive processes of metacognitive control and working memory, which are considered to be mediated by the frontal cortex and its circuitry. We studied the efficiency with which verbal items of greater value are selectively learned from among items varying in value in 14 children ages 8 15 years who had sustained severe traumatic brain injury (TBI) and in 39 typically developing age-matched children. We hypothesized that children with TBI would be disproportionately compromised in selective learning efficiency in contrast to memory span when compared to normally developing children. The results supported our hypothesis, as children with TBI performed significantly worse than controls on a measure of selective learning efficiency, but the two groups performed similarly on a measure of word recall within the same task. Furthermore, the effect of TBI on performance was demonstrated to take place at the time of encoding, rather than at retrieval. PMID- 12638065 TI - Profiles of everyday executive function in acquired and developmental disorders. AB - Executive function profiles were examined within and between several clinical disorders via the multi-domain Behavior Rating Inventory of Executive Function (BRIEF; Gioia, Isquith, Guy, & Kenworthy, 2000). Parent ratings of children with Inattentive and Combined types of Attention-Deficit/Hyperactivity Disorder (ADHD I, ADHD-C), Autistic Spectrum Disorders (ASD), moderate and severe Traumatic Brain Injury (TBI), and Reading Disabilities (RD) were compared with controls. Profile analysis revealed significant (p<.01) and substantial (eta2>.10) differences in global elevations and in profile of scale elevations between diagnostic groups. ASD, ADHD-I and ADHD-C groups exhibited greater elevations across the BRIEF scales than did RD and Severe TBI groups, who were in turn more elevated than Moderate TBI and Control groups. The ADHD-C group was unique in its frequency and severity of inhibitory deficits, while the ASD group was distinguishable by its deficits in flexibility. Within diagnostic groups, relative risk for executive dysfunction was calculated with variability present in the frequency of clinically elevated scales. While the BRIEF captures executive profiles characteristic of specific disorders in the clinical setting, it is not diagnostic in its own right and is best used within the context of a broad based evaluation. PMID- 12638066 TI - Attention and executive function deficits in adolescent sex offenders. AB - Thirty male adolescent sex offenders and 20 age-matched male adolescents completed an extensive battery of attention and executive function tests. Controls were obtained from adolescents from a socially and economically deprived background, typical of the offending group. The attention battery was based on Mirksy, Anthony, Duncan, Ahearn, and Kellam (1991) and the executive function battery on Kelly (2000a). Successful matching for IQ was not achieved and therefore ANCOVA comparisons were made between the groups, with IQ as the covariate. In attention a highly significant difference was found on the focus execute factor and a significant difference on the shift factor. In executive function there was a highly significant difference only on the response speed factor. In all cases better abilities were demonstrated by the control group. The importance of thorough neuropsychological investigation in the clinical assessment of this forensic group is supported. The clinical implications of neuropsychological deficits in terms of risk assessment and clinical management are discussed. PMID- 12638074 TI - Prenatal sonographic diagnosis of Klippel-Trenaunay-Weber syndrome associated with umbilical cord hemangioma. AB - Klippel-Trenaunay-Weber syndrome (KTW), also known as angio-osteohypertrophy syndrome, is a complex developmental disorder characterized by asymmetric hemi hypertrophy of limbs and trunk due to bony and soft tissue overgrowth that may extend across the midline, varicose veins, and cutaneous hemangiomata. This rare syndrome has been previously described in the prenatal period by ultrasonography, and in literature, there are quite different presentation of cases. We describe a case suggested as a KTW syndrome by prenatal ultrasonography. The sonographic appearance of an unilateral leg hypertrophy associated with irregular echolucent cystic areas was consistent with KTW syndrome. A spheric, weak-echogenic mass without any active blood flow located in the umbilical cord revealed by color Doppler ultrasonography was an unexpected finding. The postpartum examination confirmed the prenatal diagnosis. PMID- 12638075 TI - Congenital absence of the 5th digital ray and its proximal segmental structures. AB - Limb reduction anomalies involving digits or parts of limbs are not uncommon. Most cases take the form of terminal transverse absence defects involving hypoplasia or aplasia of all structures distal to a particular level on a limb with relative preservation of the more proximal tissues. Longitudinal absence defects are a more rare form of limb reduction anomalies. Structures within one or more digital rays are typically involved. More proximal segmental tissues may also be affected with the most serious cases resulting in absence of the radius or ulna. We report a case of isolated, longitudinal absence of the 5th finger and its corresponding proximal segmental structures in a newborn infant. The embryology of limb development and possible etiologies for skeletal absence defects are summarized. PMID- 12638076 TI - Evidence for the presence of neutralizing antibodies against human papillomavirus type 6 in infants born to mothers with condyloma acuminata. AB - Despite human papillomavirus type 6 or 11 (HPV6/11) being often vertically transmitted from mothers with condyloma acuminata (CA) to their infants, HPV related neonatal mucosal diseases are rare. The role of maternal anti-HPV6/11 neutralizing antibodies in preventing the vertical transmission remains to be unknown because of lack of the neutralization assay system of HPV infection. We experienced two cases of HPV6-positive CA during pregnancy. Neutralizing antibodies against HPV6 in maternal, umbilical, and infantile sera were determined using a surrogate assay system to monitor HPV6 pseudo-infections. The neutralizing antibodies were detected in maternal and umbilical sera and in serum of one of the infants tested at 5 weeks old. In the infant exposed to HPV6 at birth, viral DNA was not detectable in the oral cavity 5 weeks after birth. This is the first report to describe that neutralizing antibodies against HPV6 in mothers with CA go through the placenta and enter the circulation of their infants. These data may provide a mechanistic paradigm for the prevention of its vertical transmission. PMID- 12638077 TI - Association between inherited thrombophilias, antiphospholipid antibodies, and lipoprotein A levels and venous thromboembolism in pregnancy. AB - The objective of this study is to evaluate the association between venous thromboembolism (VTE) in pregnancy with thrombophilic factors. Thirty pregnant women with VTE were compared with 30 pregnant women matched by age and race without VTE and evaluated for risk factors and thrombophilia. Statistical analysis used two-tailed Fisher's exact test. VTE distribution was 30% in first trimester, 9% in 2nd trimester, 26% in 3rd trimester and 35% postpartum. Seventeen (57%) of VTE cases versus 2 (7%) of control cases had specific thrombophilia diagnosis ( p <0.001). The results were: Factor V Leiden mutation (27 vs. 3%) p = 0.026, MTHFR homozygous 677 mutation (10 vs. 44%) p = 0.017, prothrombin gene mutation (11 vs. 0%), protein C deficiency (7 vs. 0%), antiphospholipid antibodies (27 vs. 3%) p = 0.026, mean lipoprotein levels 49 versus 23 mg/dL, p = 0.008, mean homocysteine levels 7.8 versus 7.1 umol/L. An association is suggested between thromboembolic events in pregnancy and thrombophilia, especially Factor V Leiden gene mutation and elevated antiphospholipid antibodies. A new finding is the association of elevated lipoprotein A levels with thromboembolic events in pregnancy. PMID- 12638078 TI - Lethal neonatal carnitine palmitoyltransferase II deficiency: an unusual presentation of a rare disorder. AB - A term male newborn, appropriate for gestational age, developed hypothermia, severe cardiac dysrrhythmia, and nonoliguric hyperkalemia within 24 hours of birth. Despite the prenatal identification of cystic renal dysplasia without oligohydramnios, at birth, a solitary left leg vascular hemangioma and large palpable kidneys were the only anomalies. Gradually hypotonia, lethargy, and poor feeding developed and by 20 hours of age recurrent cardiac dysrrhythmias, myocardial dysfunction, and renal insufficiency with intermittent hyperkalemia were apparent. Episodes of apnea developed on day 7 followed by respiratory failure, recurrent cardiac dysrrhythmias, and death on day 12. Eventually laboratory and autopsy findings confirmed the diagnosis of lethal neonatal carnitine palmitoyltransferase II deficiency. PMID- 12638079 TI - Unexpected versus anticipated cystic periventricular leukomalacia. AB - The objective of this study is to contrast the clinical profile of premature infants who develop cystic periventricular leukomalacia (cPVL) following a severe hemodynamic event (anticipated) with infants whose cPVL is not preceded by such an event (unexpected). The study design is a Retrospective Cohort. The study setting is a Neonatal Intensive Care Unit. Infants born at a gestational age of 23-32 weeks received serial cranial ultrasounds to screen for cPVL. Infants were divided into two groups based upon whether or not they had experienced a hemodynamic event of sufficient severity to potentially cause cPVL. Details of the clinical profile of these two groups were compared. Cystic PVL occurred in 27 infants (2.1%). In 15 infants (56%) cPVL was antipicated as there was at least one severe preceding hemodynamic event. Twelve infants (44%) had not experienced such an event and the discovery of cPVL on routine cranial ultrasound was unexpected. These infants were more likely to have been born to mothers who had received tocolytics (67 versus 13%, p = 0.004), prenatal steroids (67 vs. 27%, p = 0.038), and intrapartum antibiotics (92 vs. 20%, p < 0.001) and were more likely to have been delivered more than 6 hours after arrival to the hospital (83 vs. 40%; p = 0.038) than infants whose cPVL was anticipated. The diagnosis of clinical chorioamnionitis was also more common in the group of infants whose cPVL was unexpected (50 vs. 13%, p = 0.038). Almost half (44%) of our cases of cPVL occurred unexpectedly in infants who did not appear to have an obvious cause, as they had not experienced a severe hemodynamic event. These infants were more commonly delivered to mothers who had been hospitalized long enough to have received a variety of prenatal therapies and more often had a diagnosis of chorioamnionitis. PMID- 12638080 TI - Pregnant women with the sickle cell trait are not at increased risk for developing preeclampsia. AB - The primary objective of this study was to determine whether having the sickle cell trait is independently associated with preeclampsia. We performed a retrospective cohort study of 1998 pregnant patients who either did or did not have the sickle cell trait. All patients were screened for the sickle trait using the "Sickledex" test. Data on neonatal and maternal outcome, including preeclampsia, and potential confounding variables were abstracted from medical records. Unadjusted, stratified, and multiple logistic regression analyses were used to identify interactions, and confounding between multiple variables and the association between sickle cell trait and preeclampsia. With an anticipated 6.5% rate of preeclampsia, and alpha = 0.05, this cohort study has 80% power to detect a relative risk (RR) of 2.3 for preeclampsia. Univariate analysis revealed that the two cohorts were similar with regard to primiparity, maternal age, chronic diseases, birth weight, and gestational age at delivery, but the sickle cell trait cohort was more likely to have gestational diabetes and had a higher mean body mass index (BMI). In the univariate analysis, the sickle cell trait cohort was not at increased risk for preeclampsia [unadjusted RR = 0.5, 95% CI (0.2 1.6)]. After controlling for potential confounding variables with logistic regression analysis, sickle trait was not independently associated with preeclampsia [adjusted RR = 0.5, 95% CI (0.2- 1.6)]. In contrast to prior work, these data suggest that the sickle cell trait is not an independent risk factor for preeclampsia or postpartum complications. In fact, the data are more consistent with the sickle trait being protective for developing preeclampsia. PMID- 12638081 TI - Risk factors for neonatal thrombocytopenia in preterm infants. AB - The objective of this study is to identify the risk factors for neonatal thrombocytopenia among preterm infants. During a 4-year study period all consecutive, singleton preterm deliveries (between 27 and 35 weeks of gestation) were evaluated, and separate cohorts were compared-growth restricted (small-for gestational-age; SGA) and appropriately grown (appropriate-for-gestational-age; AGA) infants. An initial comparison was done for the presence of thrombocytopenia (platelet count below 150,000/mL) and marked thrombocytopenia (below 100,000/mL). Following that, a comparison was made between the groups as determined by platelet count for various possible risk factors. Three hundred and five preterm infants were included in the study. Mean platelet count was significantly lower in the SGA group (p = 0.0009). Ninety-three neonates (31%) were thrombocytopenic and 212 infants with a normal platelet count served as controls. In the thrombocytopenic group, the rate of preeclampsia was significantly higher (p = 0.002). Thrombocytopenic infants had a significantly lower average gestational age at delivery (p = 0.002), lower birth weight (p = 0.0001), and low 5-minute Apgar score (p = 0.0002). They were more likely to suffer from intraventricular hemorrhage (IVH) ( p = 0.04) and sepsis (p = 0.002). Growth restriction, lower gestational age and low 5-minute Apgar score (<7) were found to be significantly independent risk factors for marked thrombocytopenia, when analyzed separately. Growth restriction, lower gestational age at delivery, and low 5-minutes Apgar score are significantly associated with neonatal thrombocytopenia in preterm infants, which may lead to significant morbidity. Screening these high-risk groups for thrombocytopenia might be beneficial in terms of early diagnosis and management. PMID- 12638082 TI - Significant linkage of Parkinson disease to chromosome 2q36-37. AB - Parkinson disease (PD) is the second most common neurodegenerative disorder, surpassed in frequency only by Alzheimer disease. Elsewhere we have reported linkage to chromosome 2q in a sample of sibling pairs with PD. We have now expanded our sample to include 150 families meeting our strictest diagnostic definition of verified PD. To further delineate the chromosome 2q linkage, we have performed analyses using only those pedigrees with the strongest family history of PD. Linkage analyses in this subset of 65 pedigrees generated a LOD score of 5.1, which was obtained using an autosomal dominant model of disease transmission. This result strongly suggests that variation in a gene on chromosome 2q36-37 contributes to PD susceptibility. PMID- 12638085 TI - [Psychotherapy of somatoform disorders]. AB - OBJECTIVES: To present a review of the current state of the art of psychotherapy of somatoform disorders from a psychodynamic-interpersonal perspective. METHODS: The review is based on an analysis of the literature used for evidence-based guidelines on the treatment of somatoform disorders and on our own experiences and models of these disorders. An elaboration of practical aspects of psychotherapy with these patients makes up an essential part of the review. RESULTS AND DISCUSSION: Psychotherapy of somatoform disorders is efficient and effective if the typical interpersonal patterns of these patients are taken into account and if the therapist maintains a more active attitude, avoiding early psychosocial interpretations of the somatoform complaints. PMID- 12638084 TI - Fragile X premutation tremor/ataxia syndrome: molecular, clinical, and neuroimaging correlates. AB - We present a series of 26 patients, all >50 years of age, who are carriers of the fragile X premutation and are affected by a multisystem, progressive neurological disorder. The two main clinical features of this new syndrome are cerebellar ataxia and/or intention tremor, which were chosen as clinical inclusion criteria for this series. Other documented symptoms were short-term memory loss, executive function deficits, cognitive decline, parkinsonism, peripheral neuropathy, lower limb proximal muscle weakness, and autonomic dysfunction. Symmetrical regions of increased T2 signal intensity in the middle cerebellar peduncles and adjacent cerebellar white matter are thought to be highly sensitive for this neurologic condition, and their presence is the radiological inclusion criterion for this series. Molecular findings include elevated mRNA and low-normal or mildly decreased levels of fragile X mental retardation 1 protein. The clinical presentation of these patients, coupled with a specific lesion visible on magnetic resonance imaging and with neuropathological findings, affords a more complete delineation of this fragile X premutation-associated tremor/ataxia syndrome and distinguishes it from other movement disorders. PMID- 12638083 TI - Combined analysis of genome scans of dutch and finnish families reveals a susceptibility locus for high-density lipoprotein cholesterol on chromosome 16q. AB - Several genomewide screens have been performed to identify novel loci predisposing to unfavorable serum lipid levels and coronary heart disease (CHD). We hypothesized that the accumulating data of these screens in different study populations could be combined to verify which of the identified loci truly harbor susceptibility genes. The power of this strategy has recently been demonstrated with other complex diseases, such as inflammatory bowel disease and asthma. We assessed the largely unknown genetic background of CHD by investigating the most common dyslipidemia predisposing to CHD, familial combined hyperlipidemia (FCHL), affecting 1%-2% of Western populations and 10%-20% of families with premature CHD. To be able to perform a combined data analysis, we unified the diagnostic criteria for FCHL and its component traits and combined the data from two genomewide scans performed in two populations, the Finns and the Dutch. As a result of our pooled data analysis, we identified three chromosomal regions, on chromosomes 2p25.1, 9p23, and 16q24.1, exceeding the statistical significance level of a LOD score >2.0. The 2p25.1 region was detected for the FCHL trait, and the 9p23 and 16q24.1 regions were detected for the low high-density lipoprotein cholesterol (HDL-C) trait. In addition, the previously recognized 1q21 region also obtained additional support in the other study sample, when the triglyceride trait was used. Analysis of the 16q24.1 region resulted in a statistically significant LOD score of 3.6 when the data from Finnish families with low HDL-C were included in the analysis. To search for the underlying gene in the 16q24.1 region, we investigated a novel functional and positional candidate gene, helix/forkhead transcription factor (FOXC2), by sequencing and by genotyping of two single-nucleotide polymorphisms in the families. PMID- 12638087 TI - [Social network and social support in patients with chronic inflammatory bowel disease]. AB - OBJECTIVES: The aim of the present study was to make a precise analysis of social network and social support to patients with chronic inflammatory bowel diseases (CEDE) in comparison with healthy controls. METHODS: 35 patients with colitis ulcerosa and 47 patients with Morbus Crohn were interviewed using the questionnaire for investigating social network and social support (SONET). Medical data on disease activity were also assessed. CEDE patients were compared with healthy controls, and patients with active disease were compared with those whose disease was in remission. RESULTS: In comparison to healthy controls, the patients demonstrated a significantly smaller network, comprising fewer extra familial persons but more relationships to the closest core family members. CEDE patients also reported a significantly smaller number of persons from whom they received social support. Patients with active disease were generally more unsatisfied with the support received than those whose disease was in remission. CONCLUSIONS: Our study shows that the closest family members are usually responsible for the patient's social support. Further investigation is necessary to assess the needs of the core family in this stressful situation. PMID- 12638086 TI - [Coping and appraisal in rheumatoid arthritis and fibromyalgia patients]. AB - OBJECTIVES AND METHODS: In a cognitive framework appraisal and coping are seen as major factors in the adaptation of chronic pain patients. This study evaluates the differences between rheumatoid arthritis (N = 70) and fibromyalgia (N = 74) outpatients in regard to coping and appraisal, and the relationship between appraisal, coping and adaptation in general. RESULTS: In rheumatoid arthritis patients acceptance of illness and cognitive-reappraising coping prevails. In fibromyalgia patients, however, passive, emotion-focused coping and the judgement of illness as a "threat" or "punishment" (Lipowsky) was pronounced. A connection between individual appraisal of the illness and means of coping was observed. There was also a strong relationship between coping and adaptation, with one third of the variance of the mood variables explained by appraisal and coping. The presence of a psychic disorders played only a minor role. CONCLUSIONS: A detailed understanding of the relationship between appraisal, coping, and adaptation may contribute to improved treatment concepts in pain patients. One aim of psychotherapy in chronic pain patients should be to reduce passive, emotion-focused coping and to change maladaptative concepts of illness. PMID- 12638088 TI - [Predictors of quality of life after orthopedic treatment of lower back pain due to lumbar intervertebral disc disorders]. AB - OBJECTIVES: To investigate the relevance of somatic, psychic and psychosocial factors on the health-related quality of life at the one year follow-up of patients with lower back pain. METHODS: Prospective cohort study of 109 patients recruited consecutively. At baseline and at one year follow-up self-report instruments were administered to evaluate health-related quality of life (SF-36), psychic or psychological distress (SF-36), and coping strategies (FKV-LIS). RESULTS: In regards to the physical and mental dimensions of the quality of life at follow-up, psychosocial factors evaluated at baseline were far more relevant. Using a multiple regression analysis we were able to account for 38 % of the variance in the physical dimension of the quality of life and 45 % of the variance in the mental dimension. In these two dimensions the factors "psychic distress" (GSI, SCL-90-R) and "sick leave" were significant predictors, in the mental dimension additionally "doctor shopping". Beyond that, impaired health related quality of life at baseline as well as at follow-up was related to depressive coping (FKV-LIS). CONCLUSIONS: The study shows the high impact of psychic and psychosocial factors on health-related quality of life in patients with chronic lower back pain. PMID- 12638090 TI - [Qualitative psychotherapy research in German speaking countries]. AB - OBJECTIVES: The subject of this paper is a review of qualitative research in psychotherapy and psychosomatic medicine in German speaking countries. METHODS: A short systematic survey of the relevant topics is given. RESULTS: The methodology and methods are reviewed as well as subjects and results, e.g. first interview and diagnostics, psychosomatic disorders, research into the course of psychotherapy, patient-therapist-interaction, and follow-up investigations. DISCUSSION: We briefly examine the historical and sociological backgrounds most relevant for the development of qualitative psychotherapy research. PMID- 12638089 TI - [Individualized therapy goals and treatment efficacy--a follow-up study one year after inpatient psychotherapy]. AB - Quality management of inpatient psychotherapy must integrate the different aspects of process-quality management as well as the evaluation of treatment outcome. The aim of our study was to investigate the association of the achievement of individually arranged and continuously re-evaluated therapy goals with symptomatic measures of treatment outcome (SCL-90-R, Giessener Beschwerde Bogen). 51 inpatient subjects were assessed directly before and after treatment and again at approximately 1 year after treatment (mean range 14.7 months). Therapy goal achievement was assessed on a weekly basis and, in addition, globally at the end of treatment. Pre/post-treatment results remained stabile at the follow-up assessment (SCL-90-R GSI effect size: 1.2). No significant association was found between therapy goal achievement and the degree of improvement of symptoms. This result leads us to the conclusion that the continuous evaluation of therapy goals during treatment seems to be a good instrument for quality management, but it is not suitable for the evaluation of treatment outcome. PMID- 12638091 TI - HIV infection and lipodystrophy. PMID- 12638092 TI - Echocardiography in HIV cardiac disease. PMID- 12638093 TI - Dyslipidemia in the era of HIV protease inhibitors. AB - Human immunodeficiency virus protease inhibitors are associated with metabolic abnormalities that may increase risk of atherosclerotic vascular disease, including dyslipidemia, insulin resistance, and central obesity. Dyslipidemia, characterized by hypercholesterolemia and hypertriglyceridemia, small low- and high-density lipoprotein particles, and in some cases lipoprotein(a) excess, can be severe and has been associated with endothelial dysfunction and carotid atherosclerosis. The mechanisms underlying protease inhibitor-associated dyslipidemia have not been elucidated completely, but appear to involve hepatic overproduction of very low-density lipoproteins and to a lesser extent, impaired clearance. Insulin resistance appears to mediate part of the adverse lipoprotein changes observed in patients taking protease inhibitors. Ongoing epidemiological and surrogate endpoint studies are investigating the atherogenicity of these medications. Until the risk associated with use of protease inhibitors is better understood, identifying patients at high risk for adverse vascular events such as heart attacks, cardiac death, and stroke is a high priority. This article reviews the lipid and lipoprotein abnormalities associated with use of protease inhibitors, possible mechanisms for protease inhibitor-associated dyslipidemia, its potential atherogenicity, and use of the National Cholesterol Education Program Adult Treatment Panel III Guidelines for the management of patients with dyslipidemia. PMID- 12638094 TI - Mitochondrial DNA replication, nucleoside reverse-transcriptase inhibitors, and AIDS cardiomyopathy. AB - Nucleoside reverse-transcriptase inhibitors (NRTIs) in combination with other antiretrovirals (HAART) are the cornerstones of current AIDS therapy, but extensive use brought mitochondrial side effects to light. Clinical experience, pharmacological, cell, and molecular biological evidence links altered mitochondrial (mt-) DNA replication to the toxicity of NRTIs in many tissues, and conversely, mtDNA replication defects and mtDNA depletion in target tissues are observed. Organ-specific pathological changes or diverse systemic effects result from and are frequently attributed to HAART in which NRTIs are included. The shared features of mtDNA depletion and energy depletion became key observations and related the clinical and in vivo experimental findings to inhibition of mtDNA replication by NRTI triphosphates in vitro. Subsequent to those findings, other observations suggested that mitochondrial energy deprivation is concomitant with or the result of mitochondrial oxidative stress in AIDS (from HIV, for example) or from NRTI therapy itself. PMID- 12638095 TI - Epidemiology of HIV cardiac disease. AB - The heart is an organ frequently affected in patients with acquired immune deficiency syndrome (AIDS). Since the introduction of highly active antiretroviral therapy (HAART), a sharp decline in mortality and morbidity has been observed in human immunodeficiency virus (HIV)-infected patients. However, numerous reports of myocardial infarcts in young HIV-infected patients have raised concerns of premature coronary artery disease in this population. New risk factors for coronary heart disease such as increased insulin resistance, dyslipidemia, and lipodystrophy syndrome, which are associated with HAART, may accelerate underlying arteriosclerosis in HIV-infected patients. Data on the incidence of coronary heart disease are limited to case reports and retrospective studies. Results from ongoing, large, prospective studies will provide information on whether or not HAART may increase the incidence of myocardial infarcts and whether a drastic change in HIV therapy is warranted. PMID- 12638096 TI - Structure and function of the sinus node, AV node and his bundle of the human heart: part II--function. PMID- 12638097 TI - The effect of modifying patient-preferred spinal movement and alignment during symptom testing in patients with low back pain: a preliminary report. AB - OBJECTIVE: To examine the effect on symptoms of modifying patient-preferred movements and alignments of the lumbar spine during patient examination. DESIGN: Repeated-measures study in which patients with low back pain (LBP) participated in a standardized examination that included tests of symptoms with various movements and positions. SETTING: Six university-affiliated outpatient physical therapy clinics and the local community. PARTICIPANTS: Five trained physical therapists examined a total of 185 patients (102 women, 83 men; mean age, 41.89+/ 13.29 y) with LBP. The majority of patients had multiepisode, chronic LBP. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The examination included tests of symptoms with various alignments and movements in several different positions. Seven tests were designated as primary tests. Tests that increased symptoms were followed immediately by a secondary test in which (1) patient preferred lumbar spine movement was modified or (2) the lumbar spine was positioned in a neutral alignment. Patients reported the effect of the secondary test on symptoms relative to their symptoms with the primary test. Three responses were possible: symptoms increased, remained the same, or decreased. RESULTS: Eighty-three percent of the patients reported an increase in symptoms with 1 or more of the 7 primary tests. Ninety-five percent who reported an increase in symptoms with at least 1 of the primary tests reported a decrease in symptoms with 1 or more of the 7 secondary tests. The majority of patients reported a decrease in symptoms when the spinal movement or alignment was modified for 6 of the 7 secondary tests. CONCLUSIONS: Modifying the symptom provoking movements and alignments of the spine during symptom testing resulted in a decrease in symptoms for the majority of patients. Information about specific modifications that provide relief of LBP symptoms is important because it can be used to design a treatment program that focuses on training a patient to modify the same movements and alignments in their everyday activities. PMID- 12638098 TI - Limb activation effects in hemispatial neglect. AB - OBJECTIVE: To assess the efficacy of passive and active limb movement to improve visual scanning in patients with hemispatial neglect. DESIGN: Before-after trial: behavioral analyses of a case series. SETTING: Stroke rehabilitation unit in a tertiary care hospital. PARTICIPANTS: Nine individuals with right-hemisphere stroke (mean time poststroke, 19.5 mo) and left-sided neglect, as assessed by the Sunnybrook Bedside Neglect Battery. INTERVENTION: Active left limb movement (button push; n=3) or passive left limb movement (n=8) with functional electric stimulation (FES) administered during visual scanning testing. MAIN OUTCOME MEASURES: Performance on visual scanning tests involving naming of letters and numbers. RESULTS: Both active and passive movement significantly improved target detection on the left side, but not on the right side, on the visual scanning task. Positive results were seen in 2 of 3 active movement patients and 6 of 8 passive movement patients. CONCLUSIONS: Both active and FES-stimulated passive movements are potential techniques for the treatment of hemispatial neglect. PMID- 12638099 TI - Continuous low-level heatwrap therapy for treating acute nonspecific low back pain. AB - OBJECTIVE: To evaluate the efficacy of 8 hours of continuous low-level heatwrap therapy for the treatment of acute nonspecific low back pain (LBP). DESIGN: Prospective, randomized, parallel, single-blind (investigator), placebo controlled, multicenter clinical trial. SETTING: Five community-based research facilities. PARTICIPANTS: Two-hundred nineteen subjects, aged 18 to 55 years, with acute nonspecific LBP. INTERVENTION: Subjects were stratified by baseline pain intensity and gender and randomized to one of the following groups: evaluation of efficacy (heatwrap, n=95; oral placebo, n=96) and blinding (oral ibuprofen, n=12; unheated back, wrap n=16). All treatments were administered for 3 consecutive days with 2 days of follow-up. MAIN OUTCOME MEASURES: Primary: day 1 mean pain relief (0- to 5-point verbal response scale). Secondary: muscle stiffness (101-point numeric rating scale), lateral trunk flexibility (fingertip floor distance), and Roland-Morris Disability Questionnaire over 3 days of treatment and 2 days of follow-up. RESULTS: Heatwrap therapy was shown to provide significant therapeutic benefits when compared with placebo during both the treatment and follow-up period. On day 1, the heatwrap group had greater pain relief (1.76+/-.10 vs 1.05+/-.11, P <.001), less muscle stiffness (43.1+/-1.21 vs 47.6+/-1.21, P=.008), and increased flexibility (18.6+/-.44 cm vs 16.5+/-.45 cm, P=.001) compared with placebo. Disability was also reduced in the heatwrap group (5.3 vs 7.4, P=.0002). Adverse events were mild and infrequent. CONCLUSION: Continuous low-level heatwrap therapy was shown to be effective for the treatment of acute, nonspecific LBP. PMID- 12638100 TI - Overnight use of continuous low-level heatwrap therapy for relief of low back pain. AB - OBJECTIVE: To evaluate of the efficacy and safety of 8 hours of continuous, low level heatwrap therapy administered during sleep. DESIGN: Prospective, randomized, parallel, single-blind (investigator), placebo-controlled, multicenter clinical trial. SETTING: Two community-based research facilities. PARTICIPANTS: Seventy-six patients, aged 18 to 55 years, with acute, nonspecific low back pain. INTERVENTIONS: Subjects were stratified by baseline pain intensity and gender and randomized to one of the following treatments: evaluation of efficacy (heatwrap, n=33; oral placebo, n=34) or blinding (unheated wrap, n=5; oral ibuprofen, n=4). All treatments were administered for 3 consecutive nights with 2 days of follow-up. MAIN OUTCOME MEASURES: Primary: morning pain relief (hour 0) on days 2 through 4 (0-5-point verbal response scale). Secondary: mean daytime pain relief score (days 2-4, hours 0-8), mean extended pain relief score (day 4, hour 0; day 5, hour 0), muscle stiffness, lateral trunk flexibility, and disability (Roland-Morris Disability Questionnaire). RESULTS: Heatwrap therapy was significantly better than placebo at hour 0 on days 2 through 4 for mean pain relief (P=.00005); at hours 0 through 8 on days 2 through 4 for pain relief (P<.001); at hour 0 on day 4 and at hour 0 on day 5 for mean pain relief (P<.001); on day 4 in reduction of morning muscle stiffness (P<.001); for increased lateral trunk flexibility on day 4 (P<.002); and for decreased low back disability on day 4 (P=.005). Adverse events were mild and infrequent. CONCLUSIONS: Overnight use of heatwrap therapy provided effective pain relief throughout the next day, reduced muscle stiffness and disability, and improved trunk flexibility. Positive effects were sustained more than 48 hours after treatments were completed. PMID- 12638101 TI - An assessment of gait and balance deficits after traumatic brain injury. AB - OBJECTIVE: To assess the sensations of instability that many patients report after traumatic brain injury (TBI). DESIGN: A controlled study. SETTING: A motion analysis and vestibular and balance laboratory. PARTICIPANTS: Twenty subjects, 10 with TBI and complaints of instability, and 10 without TBI. INTERVENTIONS: Balance and gait analysis. MAIN OUTCOME MEASURES: Dizziness Handicap Inventory (DHI), caloric irrigation, optokinetic testing, Dix-Hallpike Test, posturography, and center of mass (COM) movement. RESULTS: Subjects were well matched in terms of age, height, weight, and gender. DHI scores of those with and without TBI differed significantly (32.2+/-23.0 vs 0.2+/-0.63, P<.001). Caloric and optokinetic circularvection testing were abnormal only in subjects with TBI (8/10 and 4/10, respectively). Benign paroxysmal positioning vertigo was present in only 3 subjects with TBI, and this either resolved spontaneously (n=1) or was successfully treated (n=2). Composite posturography scores of those with and without TBI differed significantly (69.6+/-35.8 vs 79.5+/-40.5, P=.02). Gait parameters also differed significantly between the groups (P=.05), with the subjects with TBI having lower anterior and posterior and higher medial and lateral COM displacements and velocities. CONCLUSIONS: Patients' complaints of instability after TBI may have objective correlates and may be rectifiable. Balance and gait testing in these patients is warranted. PMID- 12638102 TI - A single-blind investigation into the hypoalgesic effects of different swing patterns of interferential currents on cold-induced pain in healthy volunteers. AB - OBJECTIVE: To compare the analgesic effects of differing swing patterns of interferential current (IFC) on cold-induced pain. DESIGN: Single-blind intervention study in which subjects completed 6 cycles of the cold-induced pain test-2 pretreatment, 2 during treatment, and 2 posttreatment. SETTING: Laboratory. PARTICIPANTS: Forty healthy volunteers. INTERVENTIONS: Subjects were randomly allocated to receive 1 of 4 IFC treatment interventions: 1 integral 1, 6 integral 6, 6 wedge 6, or burst. IFC was administered for 20 minutes via 4 electrodes attached to the forearm (quadripolar application) at a strong but comfortable intensity using amplitude-modulated frequencies of 1 to 100 Hz. MAIN OUTCOME MEASURES: Change in pain threshold and self-report of pain intensity and unpleasantness from pretreatment baseline. RESULTS: Two-way repeated-measure analysis of variance found significant effects for time and group by time interaction (P<.01), but effects for groups failed to reach statistical significance (P=0.1). This suggests that when all groups are considered together, subjects experience a rise in pain threshold when IFC devices are switched on but not when they are switched off. However, the rise in pain threshold was not dependent on the swing pattern employed. Analysis of pain intensity and unpleasantness ratings found no effects for group or group by time interaction. CONCLUSIONS: There were no differences in the hypoalgesic effects of different swing patterns. PMID- 12638103 TI - A typology of alcohol use patterns among persons with recent traumatic brain injury or spinal cord injury: implications for treatment matching. AB - OBJECTIVE: To describe empirically valid and clinically meaningful types of alcohol use among persons with recent traumatic brain or spinal cord injury. DESIGN: Cross-sectional cohort survey. SETTING: Acute inpatient rehabilitation program in a level I trauma center. PARTICIPANTS: A total of 218 (87%) of 250 consecutive initial admissions who met inclusion criteria and completed interviews. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Alcohol and drug use questionnaires, alcohol problem questions, admission toxicology results, readiness to change, and treatment preference questions. RESULTS: Participants were on average 37 years old, 84% were men, and 82% were white. Four types were identified by using k-means cluster analysis based on preinjury alcohol consumption, alcohol problems, and alcohol dependence. Cluster groups differed on extrinsic variables such as drug use, readiness to change, and interest in treatment or in attending Alcoholics Anonymous. The 4 types corresponded to those with a history of (1) alcohol abuse; (2) alcohol dependence; (3) alcohol dependence in remission, partial remission, or relapsed; and (4) normal or nondrinkers. CONCLUSION: More effective care may be possible if clinicians match common patient types to specific interventions such as education, motivational interventions, formal substance abuse treatment, and relapse prevention. PMID- 12638105 TI - Effect of fatigue on torque output and electromyographic measures of trunk muscles during isometric axial rotation. AB - OBJECTIVES: To examine the changes in torque output resulting from fatigue, as well as changes in electromyographic measures of trunk muscles during isometric axial rotation and to compare these changes between directions of axial rotation. DESIGN: Subjects performed fatiguing right and left isometric axial rotation of the trunk at 80% of maximum voluntary contraction while standing upright. SETTING: A rehabilitation center. PARTICIPANTS: Twenty-three men with no history of back pain. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Surface electromyographic signals were recorded from 6 trunk muscles bilaterally. The primary torque in the transverse plane and the coupling torques in sagittal and coronal planes were also measured. RESULTS: During the fatiguing axial rotation contraction, coupling torques of both sagittal and coronal planes were slightly decreased and no difference was found between directions of axial rotation. Decreasing median frequency and an increase in electromyographic amplitude were also found in trunk muscles with different degrees of changes in individual muscles. There were significant differences (P<.05) between right and left axial rotation exertions in median frequency slope of external oblique, internal oblique, latissimus dorsi, and iliocostalis lumborum muscles, but no such difference was found in median frequency slope of rectus abdominis and multifidus muscles. This could be attributed to different functional roles among the muscles. Similar differences (P<.05) between right and left axial rotation in median frequency slope were also detected in the electromyographic amplitude slope of the trunk muscles. Coefficient of variation of the torque output and electromyographic activation in most of the trunk muscles increased during the fatigue process. CONCLUSION: The changing coupling torque, different fatigue rate, and activation changes of trunk muscles, as well as the increase in variability during fatiguing axial rotation exertion, could affect the internal loading and stability of the spine; this needs to be further quantified in future studies. PMID- 12638104 TI - Analyzing risk factors for late posttraumatic seizures: a prospective, multicenter investigation. AB - OBJECTIVES: To ascertain the natural history and to stratify risks for the development of late posttraumatic seizures in individuals with moderate to severe traumatic brain injury (TBI). DESIGN: Prospective, observational study of individuals with TBI admitted to 4 trauma centers within 24 hours of injury. SETTING: Four tertiary care trauma centers in urban areas. PARTICIPANTS: A total of 647 individuals (>/=16 y) with any of the following abnormal computed tomography (CT) scan findings: extent of midline shift and/or cisternal compression or presence of any focal pathology (eg, punctate, subarachnoid, or intraventricular hemorrhage; cortical or subcortical contusion; extra-axial lesions) during the first 7 days postinjury or best Glasgow Coma Scale (GCS) score of .90) that combined calibration was feasible. Means and other distribution characteristics differed minimally between the 6- and 13-item estimates. The person and item separations and reliabilities were somewhat lower and the mean measurement errors somewhat higher for the 6-item estimates, but only marginally so. ICCs between 6- and 13-item estimates were .95 or higher. CONCLUSION: CAT can be used to reduce data collection time; the level of precision of estimates is minimally less than that provided by traditional assessment approaches. PMID- 12638108 TI - A prehabilitation program for physically frail community-living older persons. AB - OBJECTIVES: To describe the development and implementation of a preventive, home based physical therapy program (PREHAB) and to provide evidence for the safety and interrater reliability of the PREHAB protocol. DESIGN: Demonstration study. SETTING: General community. PARTICIPANTS: Ninety-four physically frail, community living persons, aged 75 years or older, who were randomized to the PREHAB program in a clinical trial. INTERVENTIONS: The PREHAB program built on the physical therapy component of 2 previous home-based protocols. A total of 223 assessment items were linked to 28 possible interventions, including progressive balance and conditioning exercises, by using detailed algorithms and decisions rules that were automated on notebook computers. MAIN OUTCOMES MEASURES: The percentages of participants who were eligible for and who completed each intervention, the extent of progress noted in the balance and conditioning exercises, adherence to the training program, and adverse events. RESULTS: Participants who completed the PREHAB program and those who ended it prematurely received an average of 9.7 and 7.2 interventions during an average of 14.9 and 9.5 home visits, respectively. With few exceptions, the completion rate and interrater reliability for the specific interventions were high. Despite high self-reported adherence to the training program, the majority of participants did not advance beyond the initial Thera-Band level for the upper- and lower-extremity conditioning exercises, and only about a third advanced to the highest 2 levels of the balance exercises. Adverse events were no more common in the PREHAB group than in the educational control group. CONCLUSION: Our results support the feasibility and safety of the PREHAB program, but also show the special challenges and pitfalls of such a strategy when it is implemented among persons of advanced age and physical frailty. PMID- 12638109 TI - Seat-interface pressure: a pilot study of the relationship to gender, body mass index, and seating position. AB - OBJECTIVES: To investigate the relationship between interface pressure and gender, body mass index (BMI), and seating positions, and to evaluate the implications for clinical practice. DESIGN: Group design. SETTING: Pressure mapping laboratory. PARTICIPANTS: Sixty-three student volunteers (44 women, 19 men; mean age, 22.2+/-5.1y). INTERVENTIONS: Seated-interface pressure was measured using the Force Sensing Array pressure mapping system. Seating positions used included recline (10 degrees, 20 degrees, 30 degrees ), foot support, and foot elevation. MAIN OUTCOME MEASURES: Interface pressure in the form of both average pressure (mean of the pressure sensor values) and maximum pressure (highest individual sensor value). RESULTS: Both average and maximum pressures were independent of gender, while average pressure had a significant positive correlation with BMI (r=.381, alpha=.01 level). Recline of the chair by 30 degrees significantly reduced average pressure (P<.001), whereas recline by 10 degrees or 20 degrees had no significant effect. Recline by 10 degrees, 20 degrees, and 30 degrees did not significantly alter maximum pressure. Elevation of the feet on a footstool reduced average pressure to a level approaching statistical significance, while supporting participants' feet versus leaving them unsupported significantly increased average pressure at a 0 degrees, 10 degrees, 20 degrees, and 30 degrees recline (P<.01) and maximum pressure at 0 degrees, 10 degrees (P<.01), and 30 degrees (P<.05). CONCLUSIONS: Elevating clients' feet and reclining their chair by 30 degrees reduced interface pressure and the associated risk of pressure ulcer development. Additional research is required, however, to replicate this study with participants at increased risk of pressure ulcer development. PMID- 12638110 TI - Characteristics of effective and efficient rehabilitation programs. AB - OBJECTIVE: To investigate the characteristics of rehabilitation hospitals and units correlated with gains in motor and cognitive function, after adjusting for case severity of the patients admitted and for length of stay (LOS). DESIGN: The Uniform Data System for Medical Rehabilitation (UDSMR) database was first analyzed to develop a method of adjusting for patient case severity on admission. Rehabilitation programs were surveyed to assess characteristics commonly thought to be associated with efficiency and effectiveness. Data on these characteristics were linked to UDSMR data on patient characteristics and functional gain. SETTING: Seventy-seven rehabilitation hospitals across the United States. PARTICIPANTS: A total of 37,692 inpatients from the participating rehabilitation hospitals. INTERVENTION: Comprehensive rehabilitation programs not altered by researcher. MAIN OUTCOME MEASURES: Program effectiveness was estimated by gains in motor and cognitive subscale scores of the FIM trade mark instrument between admission and discharge, adjusted for indicators of caseload severity at admission. Efficiency was estimated by adjusting gains for LOS as well. RESULTS: Primary factors affecting both motor and cognitive gains included admission function (treated curvilinearly), age, certain diagnostic distinctions, onset admission interval, admission class, and LOS. Correlations between staffing intensity and numerous other program characteristics with functional gain were meager, each accounting for less than 2% of variance. LOS was predicted by a number of factors, notably by the percentage of managed care cases (r=-.20), but not by staffing intensity. CONCLUSIONS: Relationships between rehabilitation practices and functional gains by patients do not appear to be simple or overt. Continued research is needed to identify reliable connections between rehabilitative processes and patient outcomes in practice. PMID- 12638111 TI - Repetitive stretch induces c-fos and myogenin mRNA within several hours in skeletal muscle removed from rats. AB - OBJECTIVE: To investigate what effect repetitive stretch has on gene transcription in muscle tissues removed from rats. DESIGN: Case-control study. SETTING: University laboratory. ANIMALS: Eight female 8-week-old Wistar rats. INTERVENTIONS: Rat anterior tibial and soleus muscles were removed and repetitively stretched about 20% of the muscle length by using the Magnus method in Krebs-Ringer solution. Stretch-relax cycles 15 times per minute were continued for 2 hours in 1 group and 4 hours in another group. Contralateral muscles removed from the same rat were incubated without stretch as the control. MAIN OUTCOME MEASURES: After repetitive stretch, total ribonucleic acid (RNA) was extracted from the muscles and Northern blotting was carried out using c-fos and myogenin oligonucleotide as probes. RESULTS: Repetitive stretch during 2 hours significantly increased the expression of c-fos messenger RNA (mRNA) in both the tibialis anterior and soleus muscles. Increased expression of myogenin mRNA in both muscles was observed with repetitive stretch for 4 hours. Because of the initial increase of immediate early gene c-fos mRNA and the subsequent increase of myogenic transcription factor myogenin mRNA, it appears that repetitive muscle stretch induces sequential progression of gene transcription toward muscle hypertrophy. CONCLUSIONS: Mechanical repetitive stretch for a few hours would be one of the stimulating factors for muscle growth and should be available for muscle training in clinical rehabilitation. PMID- 12638112 TI - Analyses of gait, reaching, and grasping in children after traumatic brain injury. AB - OBJECTIVE: To evaluate motor behavior in children after traumatic brain injury (TBI) with quantitative instrumented measures of gait and of functional hand movements (reaching, grasping) and with clinical assessments. DESIGN: Case control study. SETTING: Tertiary pediatric trauma rehabilitation center in Germany. PARTICIPANTS: Twenty children (age range, 6-13 y) with moderate or severe TBI were examined 1+/-1.2 years (mean +/- standard deviation) postinjury. Fifteen were reexamined 2 months later. Control data were obtained from 20 healthy children and matched for age, gender, and school grade. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Quantitative measures included 10 spatiotemporal gait parameters and 6 variables describing reaching and grasping. Qualitative scores of gait and upper-limb movements were also obtained. RESULTS: Gait velocity and step and stride lengths were significantly smaller in children after TBI than in control subjects (Mann-Whitney U test, P<.05). Reach-to-grasp movements of the TBI children were characterized by a significantly longer reaction time (Mann-Whitney U test, P<.05) and movement duration, reduced velocity, and coordination deficits. The instrumented measures did not change significantly in 2 months. Several significant correlations between clinical and instrumented measures were obtained. CONCLUSION: Functional motor behavior is affected in children after moderate or severe TBI. To supplement clinical assessments with objective data, impairments of gait, reaching, and grasping movements can be recorded with instrumented measures. PMID- 12638113 TI - Young traffic victims' long-term health-related quality of life: child self reports and parental reports. AB - OBJECTIVES: To describe the long-term health-related quality of life (HRQOL) reported by young traffic injury victims and to assess the child-parent agreement on the child's HRQOL. DESIGN: Cohort study with a mean follow-up of 2.4 years. SETTING: Traumatology department in a university hospital in The Netherlands. PARTICIPANTS: All traffic injury victims treated at the traumatology department in 1996-1997 and aged 8 to 15 years at follow-up (N = 254). The data of 157 child parent pairs were available for analysis (mean follow-up age, 12+/-2.4 y; 57% boys; 24% hospitalized). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: TNO AZL Children's Quality of Life parent and child questionnaires. RESULTS: Young traffic injury victims reported a significantly lower HRQOL in the motor and autonomy scales compared with contemporaries in the reference group. The child parent agreement ranged from low to moderate (intraclass correlations,.35-.67). A comparison made between the children and their parents found that the children were more negative regarding the physical complaints and the motor, autonomy, and positive-emotion scales. CONCLUSIONS: Young traffic injury victims reported a reasonably good long-term HRQOL, and, surprisingly, few psychologic problems were revealed. Physicians who rely only on parental reports may overestimate the child's HRQOL, especially when assessing the physical functioning. The child's own reports should not be neglected in the assessment of a comprehensive picture of the child's HRQOL. PMID- 12638114 TI - Training with an upper-limb prosthetic simulator to enhance transfer of skill across limbs. AB - OBJECTIVE: To examine what effect bilateral transfer of movement across limbs may have in a person's ability to learn use of an upper-limb prosthetic simulator. DESIGN: Randomized trial. SETTING: University laboratory. PARTICIPANTS: Able bodied subjects randomly assigned to 1 of 3 groups. INTERVENTIONS: Subjects performed 3 different tasks that required manipulation of objects with the simulator. Group 1 practiced with the simulator on the preferred limb and then transferred it to the nonpreferred limb; group 2 practiced with the simulator on the nonpreferred limb and then transferred it to the preferred limb; group 3 was a control group. Groups 1 and 2 underwent pretest trials, acquisition practice, posttest trials, and a 24-hour retention test; the control group followed the same design with the exception of acquisition practice. MAIN OUTCOME MEASURES: Elapsed time (1) from a signal to move until movement began (initiation time) and (2) from the beginning of movement to task completion (movement time). RESULTS: Compared with the controls, groups 1 and 2 significantly reduced initiation time across all tasks from pretest to posttest (P=.003) and from pretest to retention test (P=.029). Groups 1 and 2 did not differ from each other. Movement time did not differ among the groups in the posttest. However, groups 1 and 2 significantly (P=.026) reduced movement time across all tasks from pretest to retention test compared with the control group. Groups 1 and 2 did not differ from each other. CONCLUSION: The effects of bilateral transfer were evident for initiation time immediately on transfer, and this learning effect persisted to the retention test. The ability to execute movement, represented by movement time, occurred after consolidation of learning was complete. Cross-limb training with a prosthetic simulator may be useful for persons with recent unilateral upper-extremity amputation who are learning to use a prosthesis. PMID- 12638116 TI - Lumbar artery pseudoaneurysm in traumatic spinal cord injury: a case report. AB - Lumbar artery pseudoaneurysm is a rare vascular complication of trauma. This case report concerns a 24-year-old man with a lumbar-level spinal cord injury (SCI) secondary to a gunshot wound who developed severe exacerbation of low back and flank pain during inpatient rehabilitation. Diagnostic investigations at an acute care hospital revealed a left lumbar artery pseudoaneurysm. This was treated by transcatheter embolization, which resulted in a marked reduction in pain. The patient resumed inpatient rehabilitation without further complications. This case report highlights the importance of early diagnosis of lumbar artery pseudoaneurysm, a potentially fatal complication that can occur in patients with traumatic lumbosacral SCI. Physiatrists should include lumbar artery pseudoaneurysm in the differential diagnosis for back, flank, or abdominal pain in this patient population. PMID- 12638115 TI - Changes in movement characteristics of the spastic upper extremity after botulinum toxin injection. AB - OBJECTIVE: To examine the longitudinal effects of botulinum toxin injection on movement characteristics of the spastic upper extremity in children by using motor control testing (MCT) techniques and standard clinical measures. DESIGN: Open-label clinical trial. SETTING: Motor control laboratory at an academic medical center. PARTICIPANTS: A convenience sample of 9 subjects (5 boys, 4 girls; age range, 7-16 y) with cerebral injury (stroke or cerebral palsy) and asymmetric upper-extremity function because of spasticity. Eight subjects had right-sided involvement. INTERVENTIONS: Botulinum toxin injection to the involved upper extremity, involving elbow, wrist, and finger flexors, depending on clinical presentation. MAIN OUTCOME MEASURES: Clinical measures included range of motion (ROM), the Ashworth Scale, FIM trade mark instrument, Pediatric Evaluation of Disability Inventory, portions of the Bruininks-Oseretsky Test of Motor Proficiency, and the Purdue pegboard. MCT consisted of visually guided reaching, bilateral finger-to-nose movements, hand tapping, and isometric pinch force tasks. Kinematic assessments were made before and at 2, 4, 6, 12, 18, and 24 weeks after botulinum toxin injection. RESULTS: All subjects had increased ROM and decreased Ashworth values throughout the testing period. In motor control tasks, improvement typically occurred earlier in the least complex movements, such as hand tapping, with 6 of 9 subjects showing a maximum, although transient, unilateral tapping speed by 6 weeks. A similar time course was observed for pinch force tasks. Improvement in more complex, forward-reaching tasks occurred much later (week 12 or later) or did not occur at all. As with the hand tasks, improved reach performance declined toward the end of the testing period. All subject showed minimal or no improvement in bilateral finger-to-nose movements. Neither maximum changes in ROM or Ashworth values correlated with improvements in functional elbow extension during sit and reach tasks, with 3 subjects with normal active ROM showing late onset or no change in reach. CONCLUSIONS: Although botulinum toxin reduced tone and increased ROM of the spastic upper extremity, the time course and degree of motor improvement appears to depend on the complexity of the task. Future research should focus on the value of adjunct therapy, such as task-specific training, in addition to botulinum toxin treatments to facilitate functional improvement of the spastic upper extremity. PMID- 12638117 TI - Upper-extremity deep vein thrombosis and paralysis: a case report. AB - Upper-extremity deep venous thrombosis (DVT) has been understudied in the rehabilitation population. Patients undergoing rehabilitation often have many risk factors that predispose them to DVT. It is important to recognize upper extremity DVT, because recent studies have shown it to pose a significant risk for pulmonary embolus and death. This is the first reported association of an upper-extremity DVT with paralysis. PMID- 12638118 TI - The importance of the complete history in the discovery of a potential suicide: a case report. AB - Suicidal ideation and completed suicides are an increasing problem among the elderly. In 1992, the elderly accounted for 13% of the population but represented 20% of all completed suicides. There are recognized risk factors for suicides in elderly patients, which include depression, deteriorating physical health, and loss of independent functioning. A complete history enables the examiner to establish a relationship with the patient and to formulate a diagnosis. Unfortunately, histories are often incomplete. Many factors can account for this, including financial pressures, patient volume, and overspecialization. The physiatric history is the integration of many parts. It incorporates not only the physiatrist's evaluation but those of other disciplines as well, for example, physical and occupational therapy. The physiatric history is a sensitive tool for assessing the state of being of the whole patient. We describe a case in which a careful and complete physiatric history and physical examination revealed an elderly patient with suicidal ideation and a plan. PMID- 12638119 TI - Applying the SOM model to text classification according to register and stylistic content. AB - We report on the application of the Self-Organizing Map (SOM) classification method to the task of categorizing texts according to their register and the style of their author. The SOM has been selected as its performance in various data-mining applications has been found to be highly successful. Here, the method is evaluated against the task of clustering textual data which are corpora of texts written in the Greek language; the parameters used depict linguistically important structural properties of the texts. The experiments reported indicate that the SOM results are equivalent to those generated by statistical methods. PMID- 12638120 TI - A hierarchical method for finding optimal architecture and weights using evolutionary least square based learning. AB - In this paper, we present a novel approach of implementing a combination methodology to find appropriate neural network architecture and weights using an evolutionary least square based algorithm (GALS).1 This paper focuses on aspects such as the heuristics of updating weights using an evolutionary least square based algorithm, finding the number of hidden neurons for a two layer feed forward neural network, the stopping criterion for the algorithm and finally some comparisons of the results with other existing methods for searching optimal or near optimal solution in the multidimensional complex search space comprising the architecture and the weight variables. We explain how the weight updating algorithm using evolutionary least square based approach can be combined with the growing architecture model to find the optimum number of hidden neurons. We also discuss the issues of finding a probabilistic solution space as a starting point for the least square method and address the problems involving fitness breaking. We apply the proposed approach to XOR problem, 10 bit odd parity problem and many real-world benchmark data sets such as handwriting data set from CEDAR, breast cancer and heart disease data sets from UCI ML repository. The comparative results based on classification accuracy and the time complexity are discussed. PMID- 12638121 TI - A local training-pruning approach for recurrent neural networks. AB - The global extended Kalman filtering (EKF) algorithm for recurrent neural networks (RNNs) is plagued by the drawback of high computational cost and storage requirement. In this paper, we present a local EKF training-pruning approach that can solve this problem. In particular, the by-products, obtained along with the local EKF training, can be utilized to measure the importance of the network weights. Comparing with the original global approach, the proposed local approach results in much lower computational cost and storage requirement. Hence, it is more practical in solving real world problems. Simulation showed that our approach is an effective joint-training-pruning method for RNNs under online operation. PMID- 12638122 TI - Text compression using hybrids of BWT and GBAM. AB - In this paper we considered a theoretical evaluation of data and text compression algorithm based on the Burrows-Wheeler Transform (BWT) and General Bidirectional Associative Memory (GBAM). A new data and text lossless compression method, based on the combination of BWT1 and GBAM2 approaches, is presented. The algorithm was tested on many texts in different formats (ASCII and RTF). The compression ratio achieved is fairly good, on average 28-36%. Decompression is fast. PMID- 12638123 TI - Global asymptotic stability of a class of dynamical neural networks. AB - The dynamics of cortical cognitive maps developed by self-organization must include the aspects of long and short-term memory. The behavior of the network is such characterized by an equation of neural activity as a fast phenomenon and an equation of synaptic modification as a slow part of the neural biologically relevant system. We present new stability conditions for analyzing the dynamics of a biological relevant system with different time scales based on the theory of flow invariance. We prove the existence and uniqueness of the equilibrium, and give a quadratic-type Lyapunov function for the flow of a competitive neural system with fast and slow dynamic variables and thus prove the global stability of the equilibrium point. PMID- 12638124 TI - mPPP1R16B is a novel mouse protein phosphatase 1 targeting subunit whose mRNA is located in cell bodies and dendrites of neurons in four distinct regions of the brain. AB - We cloned a cDNA encoding a novel mouse protein whose human homolog has been annotated in GenBank as a regulatory subunit of protein phosphatase 1, PPP1R16B. Both the primary protein sequence and the domain structure are highly conserved between PPP1R16B and proteins of unknown function from other species, such as Caenorhabditis elegans and Drosphila melanogaster. Besides a protein phosphatase 1 interaction motif, mouse PPP1R16B (mPPP1R16B) and the related proteins contain ankyrin repeats that may constitute binding sites for other proteins and C terminal prenylation signals that are likely to target the proteins to the plasma membrane. In the adult mouse, Ppp1r16b mRNA is expressed in most tissues examined, with highest expression levels in kidney and brain. In the brain, Ppp1r16b message is particularly enriched in the olfactory bulb, striatum, dentate gyrus, and cerebellum. During postnatal cerebellar development, Ppp1r16b mRNA expression levels increase gradually and are maximal around postnatal day 30. In situ hybridization revealed that Ppp1r16b message is found in both the cell bodies and the dendrites in Purkinje cells of the cerebellum and granule neurons of the dentate gyrus. PMID- 12638125 TI - The distribution of progestin receptor mRNA in rat brainstem. AB - Hypothalamic sites wherein P4, through progestin receptor, (Pgr; commonly abbreviated PR), maximizes the expression of female sexual behaviors and gonadotropin surge release have been studied intensively. However, little is known regarding PR expression in brainstem regions likely to regulate changes in autonomic functions observed when P4 levels are elevated (i.e. pregnancy). Using in situ hybridization, we found PR mRNA-containing cells widely distributed throughout the brainstem of ovariectomized, estradiol-treated Sprague-Dawley rats, with high expression in regions including the medial vestibular nucleus, nucleus of the solitary tract, substantia nigra (compact part), ventral tegmental area, hypoglossal nucleus, locus coeruleus, Purkinje cell layer of the cerebellum and inferior olivary complex. We also detected moderate to high levels of PR gene expression in several regions, such as the trapezoid nucleus, facial nucleus, periaqueductal gray regions, and rostral ventrolateral medulla. These results demonstrate that PR expression is widespread in the brainstem and identify nuclei wherein P4 may act to influence a number of physiological functions during pregnancy. PMID- 12638126 TI - mRNA expression of the murine glycoprotein (transmembrane) nmb (Gpnmb) gene is linked to the developing retinal pigment epithelium and iris. AB - The murine homologue of the human glycoprotein (transmembrane) NMB (GPNMB) gene was identified by subtractive cloning from in vitro cultured murine primary osteoblast cells and subsequent RACE-PCR. GPNMB is a highly glycosylated type I transmembrane protein that shares significant sequence homology to several melanosomal proteins. Increasing expression of Gpnmb mRNA was observed during differentiation of murine primary osteoblast cell cultures. To address the potential functions of GPNMB we analysed its mRNA-expression during murine embryonic development. In early development Gpnmb mRNA is detected at high levels in the outer layer of the retina. Later in development expression gets restricted to the retinal pigment epithelium and iris. At the cytoplasmic domain of GPNMB, a conserved di-leucin-based endosomal/melanosomal-sorting signal (ExxPLL) was located, present as well in several known melanosomal proteins. To analyse the subcellular localization we used EGFP-tagged GPNMB transfected in COS7 and HEK293 cells. In both non-pigmented cell lines, the EGFP-GPNMB fusion protein was localized to vesicular, endosomal like structures. Sequence homology to known melanosomal proteins, mRNA expression and subcellular localization are suggestive for GPNMB as an intracellular, endosomal/melanosomal compartment specific protein important for melanin biosynthesis and the development of the retinal pigment epithelium and iris. As the gene coding for human GPNMB was localized to chromosome 7p15, a locus involved in the human inherited disease cystoid macular edema, also known as dominant cystoid macular dystrophy (OMIM 153880) we highly suggest that GPNMB is a candidate gene for this human inherited disease. PMID- 12638127 TI - Localization of messenger RNAs encoding enzymes associated with malonyl-CoA metabolism in mouse brain. AB - Malonyl-CoA acts a fuel sensor in the pancreas, liver and muscle. Similarly, malonyl-CoA is implicated in satiety regulation in the brain. Expression of genes encoding enzymes implicated in regulation of malonyl-CoA levels was examined in murine brain. Acetyl-CoA carboxylase (ACC) alpha-isoform, fatty acid synthase and malonyl-CoA decarboxylase are highly expressed in the hippocampus, habenula nucleus, cerebral cortex and areas of the hypothalamus, whereas the ACC-beta isoform and liver-type carnitine palmitoyltransferase I (CPTI-L) are principally expressed in the choroid plexus. Thus different brain regions appear to be functionally configured primarily for either fatty acid synthesis or beta oxidation. Localization of transcripts encoding enzymes involved in fatty acid synthesis and beta-oxidation in distinct nuclei of the hypothalamus supports a role for malonyl-CoA as a potential effector of satiety. PMID- 12638128 TI - Distribution of serine proteinase inhibitor, clade B, member 6 (Serpinb6) in the adult mouse brain. AB - In the brain, Serpinb6 was identified as an endogenous inhibitor of neuropsin, a member of the S1 (clan SA) family of serine proteases [J. Biol. Chem. 276 (2001) 14562]. In the present study, we investigated the localization of Serpinb6 in the adult mouse brain using in situ hybridization histochemistry and immunohistochemistry. Region-specific patterns of expression were observed and two characteristics were recognized. First, the forebrain limbic area that expressed neuropsin mRNA contained Serpinb6 mRNA at moderate levels but not the lateral septum. On the other hand, Serpinb6 mRNA was also expressed moderately in the substantia nigra-ventral tegmental area system, whose fibers projected to the lateral septum. Additionally, Serpinb6 protein was detected in the lateral septum. Together, it was suggested that the expression of neuropsin in the brain is regulated entirely by Serpinb6. Second, Serpinb6 mRNA and the protein were strongly expressed in most somatic and visceral motoneurons among cranial nerve nuclei. This suggests that another serine protease is regulated by Serpinb6 in motoneurons and/or fibers. PMID- 12638129 TI - Expression pattern of the homeobox protein NKX2-1 in the developing Xenopus forebrain. AB - Although morphological data suggest that the amphibian forebrain contains similar subdivisions to those observed in birds and mammals, it is presently unclear whether the same patterning mechanisms are conserved among all three classes of tetrapods. Here we report that NKX2-1, a transcription factor that is essential for the ventral patterning of the forebrain in birds and mammals, is expressed in corresponding (homologous) domains in the developing Xenopus forebrain. NKX2-1 expression is restricted to two domains in the amphibian forebrain: (1) a ventral diencephalic domain, with expression limited to hypothalamic structures; and (2) a telencephalic domain, with expression in the medial ganglionic eminence, preoptic area and part of the septum. Thus, the detailed analysis of the distribution of NKX2-1 provides the first unequivocal evidence for distinct progenitor zones within the amphibian forebrain through embryonic and larval development. PMID- 12638130 TI - Expression of Zash-1a in the postembryonic zebrafish brain allows comparison to mouse Mash1 domains. AB - Four areas in the late embryonic murine forebrain, i.e. the subpallium (striatum), the preoptic region, the ventral thalamus, and the hypothalamus, have been described to express the basic helix-loop-helix (bHLH) gene mammalian achaete-scute homolog Mash1 (Ascl1, Mouse Genome Informatics) in a complementary fashion to another bHLH gene, neurogenin1 (ngn1) (Neurod3, Mouse Genome Informatics), which is expressed in directly adjacent forebrain regions. We report here that the four regions previously identified as subpallium, preoptic region, ventral thalamus and hypothalamus (i.e. ventral inferior lobe) in the postembryonic zebrafish brain show Zash-1a expression at 3 days postfertilization (dpf), whereas none of those areas express the bHLH gene neuroD (nrd) between 2 and 5 dpf. This indicates that two well established alternative genetic pathways involved in neurogenesis in the amniote (mammalian) brain are present in homologous phenotypic locations in the anamniote (zebrafish) brain as well and that these pathways possibly act similarly in the generation of different neuronal phenotypes (e.g. subpallial GABAergic interneurons versus pallial glutamatergic projection neurons, or dopaminergic neurons versus other neurotransmitter phenotypes). Furthermore, previous initial identification of early postembryonic brain subdivisions in the zebrafish is strongly corroborated by these expression patterns. PMID- 12638131 TI - Gene expression profile from the striatum of amphetamine-treated rats: a cDNA array and in situ hybridization histochemical study. AB - Changes in gene/protein expression markedly outlast the transient changes in behavior evoked by a single dose of a psychostimulant. These changes in gene expression are thought to underlie and/or trigger enduring changes in neuroplasticity that lead to drug addiction. We used cDNA arrays to gain a more complete picture of changes in striatal gene expression 1 and 3 h after an acute injection of amphetamine. Consistent, reliable gene expression changes were detected when criteria of at least a 1.5-fold difference and three replicate hybridizations using independent samples were performed. Using these criteria, the mRNA for three immediate early genes (IEGs), coding for activity-regulated cytoskeletal-associated protein (Arc), nerve growth factor-induced protein A (NGFI-A; early growth response protein 1) and nerve growth factor-induced protein B (NGFI-B), were upregulated 1 and 3 h after amphetamine as previously described. Novel genes, RL/IF-1 (coding for I kappa B alpha chain) and serum/glucocorticoid regulated serine/threonine protein kinase (SGK) also were increased throughout the striatum, at 1 but not 3 h. Conversely, amphetamine increased the mRNA coding for the secretogranin II precursor (chromogranin C) only at the 3 h time point when a specific decrease in regulator of G-protein signaling 4 (RGS4) mRNA was also observed. Gene changes and unique patterns of expression were verified by in situ hybridization, providing valuable information about changes in gene expression in response to acute amphetamine. PMID- 12638132 TI - Murine Lix1, a novel marker for substantia nigra, cortical layer 5, and hindbrain structures. AB - We describe the expression of Lix1 in the mouse. Starting at E8, transcripts are present in a regionalized fashion and persist throughout development. mLix1 is expressed in the cortical plate, subventricular zone, layer 5 of the postnatal cortex, the substantia nigra, dorsal root ganglia, specific nuclei of the brain stem and in spinal cord. Limb buds and facial primordia show transient expression. The prominent expression of mLix1 in the developing cerebral cortex and in the substantia nigra pars compacta makes this novel gene a candidate marker for both of these tissues. PMID- 12638134 TI - [Epidemiological study of pneumococcal infection and Haemophilus influenzae infection]. PMID- 12638133 TI - Characterization and expression of the gene encoding membralin, an evolutionary conserved protein expressed in the central nervous system. AB - We have identified a novel gene that encodes an evolutionary conserved protein that we name membralin since it contains multiple transmembrane regions. The human gene C19orf6 localizes to chromosome 19p13.3. Splice variant membralin-1 is encoded by 11 exons, translating into 620 amino acids. In addition, we found evidence for two additional splice variants in the human. The mouse gene ORF61 localizes to chromosome 10. We cloned two splice variants in mouse: membralin-1, which is encoded by 12 exons, translating into 574 amino acids, and membralin-2, which translates into 598 amino acids. The existence of rat membralin-1 (574 amino acids long) is, so far, only supported by in situ hybridization result, whereas the existence of rat membralin-2 (598 amino acids long) is strongly supported by overlapping ESTs. Gene homologues were also identified in fruit-fly (CG8405, chromosome 2R 52; two splice variants), nematode (chromosome III), and Arabidopsis thaliana (chromosome 1). Sequence analysis revealed no closely related genes, suggesting that membralin represents the sole member of a unique protein family. PMID- 12638135 TI - [Bacterial flora of upper larynx in infants in a day-care center in Japan]. PMID- 12638136 TI - [Pulse-field electrophoresis for epidemiological study on bacterial flora of nasopharynx in infants in a day-care center in Japan]. PMID- 12638137 TI - [Comparative study on bacterial flora of oral cavity and upper pharynx in healthy elderly]. PMID- 12638138 TI - [Epidemiological study on patients with pneumococcal respiratory tract infection in a Japanese health facility during the past year]. PMID- 12638139 TI - [Clinical isolates and treatment of acute otitis media]. PMID- 12638140 TI - [Therapy for children with acute pneumococcal otitis media in an outpatient clinic]. PMID- 12638141 TI - [High-dose cefditren therapy for children with acute otitis media that had Streptococcus pneumoniae and haemophilus influenzae isolated from the nasopharynx]. PMID- 12638142 TI - [Result of surveillance for clinical isolates from otorhinolaryngological infections in Japan]. PMID- 12638143 TI - [Pediatric patients with common cold together with bacterial infection of upper respiratory tract--bacterial isolates from pharynx]. PMID- 12638144 TI - [Eradication of bacteria in upper pharynx of children treated with antibiotics]. PMID- 12638145 TI - [Community-acquired infections caused by Streptococcus pneumoniae and Haemophilus influenzae(discussion)]. PMID- 12638146 TI - [Epidemiological study of pneumococcal infections and Haemophilus influenzae infections in the last three years]. PMID- 12638147 TI - [Statistical study of penicillin-resistant Streptococcus pneumoniae in otorhinolaryngologic patients in a general hospital in Japan]. PMID- 12638148 TI - [Frequency of isolation of Haemophilus influenzae in an ENT clinic in Japan and the antibiotic resistance]. PMID- 12638149 TI - [Antibacterial sensitivity of Haemophilus influenzae isolated from patients in a hospital in Japan]. PMID- 12638150 TI - [Present status of pneumococcal infection and Haemophilus influenzae infection in Fukushima Prefecture, Japan]. PMID- 12638151 TI - [Oral bacterial flora in aged persons]. PMID- 12638152 TI - [Mid term report of the CDTR clinical trial for PRSP and BLNAR infections]. PMID- 12638153 TI - [Bacteriological aspects in the mid term report of the CDTR clinical trial for PRSP and BLNAR infections]. PMID- 12638154 TI - [Comparative study on bacterial eradication rate and clinical efficacy of CDTR, CFPN, and FRPM for treatment of children with otitis media and lower respiratory tract infection due to Streptococcus pneumoniae and Haemophilus influenzae]. PMID- 12638155 TI - [Isolation of bacteria and treatment of patients with pneumococcal and Haemophilus influenzae infections]. PMID- 12638157 TI - [Infant case of acute otitis media due to PISP not eradicated with double-dose of CDTR]. PMID- 12638156 TI - [Efficacy and safety of twice-daily administration of CTDR]. PMID- 12638158 TI - [Pathological findings in temporal bone of fatal case with pneumococcal meningitis]. PMID- 12638159 TI - [Refractory acute otitis media and the risk factors]. PMID- 12638160 TI - [Pediatric patients with refractory acute otitis media due to BLNAR]. PMID- 12638162 TI - [Mode of transmission and natural course of pneumococcal infection]. PMID- 12638161 TI - [Discussions on PRSP and BLNAR]. PMID- 12638163 TI - [Transmission of pneumococcal otitis media within siblings and drug sensitivity of the pathogen]. PMID- 12638164 TI - [Study on Streptococcus pneumoniae isolated from infants in widely-separated primary care facilities in Japan]. PMID- 12638165 TI - [Frequency of isolation of PRSP and BLNAR from infants in daycare centers in Japan ]. PMID- 12638166 TI - [Low dose exposure to cadmium and its health effects. (3) Toxicity in laboratory animals and cultured cells]. AB - We have reviewed studies on cadmium (Cd) toxicity in laboratory animals and cultured cells with special attention to the disruption in cellular signal transduction, involvement in apoptosis of Cd, the cellular transport system for Cd and roles of metallothionein as a protective factor against Cd. Cd affects cellular functions by perturbing signal transductions, such as protein kinase C, mitogen-activated protein kinase and cyclic AMP pathways, but how the disruption of these pathways by Cd leads to the manifestation of toxicity in vivo is largely unknown. Exposure to cadmium at relatively high and low levels causes necrosis and apoptosis, respectively, which suggests that the mode of cell death by cadmium is dependent upon its exposure level. On the other hand, utilization of Ca2+ channels, DMT1 (divalent metal transporter 1) and a novel transport system having high-affinity for Mn2+ and Cd2+ were found to act as Cd transport systems via the cellular membrane. Metallothionein-I/II-null mice are highly susceptible to renal toxicity, hepatotoxicity, bone injury, hematotoxicity and immunotoxicity caused by chronic Cd exposure. Thus, metallothionein plays an important role in detoxification of Cd toxicity. PMID- 12638167 TI - [Low dose exposure to cadmium and its health effects. (4) Body burden, critical concentration and life expectancy]. AB - As the last manuscript in our series of review articles on cadmium (Cd) and health effects, we reviewed research articles on epidemiologic and experimental studies on exposure levels of Cd in occupational and environmental settings in various countries, disposition and body burden of Cd, critical concentrations of Cd in the kidney of humans and animals with a focus on biomarkers for renal dysfunction, and life expectancy in Cd-polluted areas and reference areas. After this manuscript was compiled, cadmium levels in rice crops received significant attention, since the risk assessment of cadmium is now under review and discussion by the Joint Expert Committee of Food Additives and Contaminants organized by the Food Agricultural Organization and World Health Organization in 2003. We hope that the information compiled in this review may provide directions for future studies on the health risk assessment of Cd. PMID- 12638168 TI - [A review of epidemiological studies about the incidence and etiological factors of cryptorchidism--relevance to endocrine-disrupting chemicals]. AB - Recently, it has been reported that the incidence of male genitourinary abnormalities (such as cryptorchidism) has been increasing because of the presence of endocrine-disrupting chemicals (EDCs) in the environment. This report reviews the papers related to cryptorchidism beginning in the 1960s to clarify the trends in incidence of cryptorchidism and to discuss the association between cryptorchidism and risk factors, including EDCs. The incidence of cryptorchidism tended to increase in almost researched countries until the mid-1980s, but has shown regional differences since 1990. In Japan, the trend of the incidence of cryptorchidism is unclear because there is no monitoring system for cryptorchidism. It is necessary to establish both criteria and a monitoring system to research the incidence of cryptorchidism. The risk factors in epidemiological studies include low birth weight, being small for gestational age, a short gestation period, high maternal body weight, the change during pregnancy of maternal hormones (by exposure to diethylstilbestrol DES or estrogen), and exposure to chemicals. However, the associations between children with cryptorchidism and exposure to EDCs as well as between cryptorchidism and exposure to estrogen of the fetus are not clear because very few reports have quantitatively evaluated EDCs. It is known that some EDCs act as estrogen receptor agonists and change hormonal mobility as exogenous estrogen. It has also been suggested that hormonal drugs such as DES affect the maternal endocrine system in pregnancy and are associated with the incidence of cryptorchidism. In the future, epidemiological studies that clarify the association between cryptorchidism and EDCs such as daidzein and genistein and the quantitative evaluation of exposure factors using biological materials are necessary. PMID- 12638169 TI - [Neurotoxicity of organophosphorus and dithiocarbamate compounds]. AB - The neurotoxicity of organophosphorus compounds (OPs) including leptophos, TOCP and triphenyl phosphite and dithiocarbamate compounds were reviewed in this study. The major neurotoxicities of OPs were acute toxicity produced by the acetylcholine esterase (AChE) inhibiting action of OPs and delayed neurotoxicity produced by such OPs as leptophos and TOCP. The direct action of OP on the muscarinic and/or nicotinic acethylcholine receptors in the synaptic membranes have lately attracted attention in relation to acute toxicity. Delayed neurotoxicity is a delayed onset of prolonged locomotor ataxia resulting from a single or repeated exposure to an OP. Although neurotoxic esterase (NTE) inhibition might be related to the onset of organophosphate-induced delayed neurotoxicity (OPIDN), the precise mode of action is not yet clear. The effect of dithiocarbamates on the nervous system is also mentioned, because the compounds are currently suspected not only for neurotoxicity, but also as endocrine disrupting chemicals. Although dithiocarbamates showed weak neurotoxicity in adult animals, we need to pay more attention to developmental neurotoxicity. PMID- 12638170 TI - [The effect of chronic stress on neutrophil function in tumor-inoculated rats]. AB - OBJECTIVES: In order to clarify the relation between stress load and inoculated tumor growth in conjunction with neutrophil functions, several kinds of stress such as physical (Ph) and psychological (Ps) stress were loaded on rats either SST-2 tumor cell inoculated or control, and the functions of their peripheral neutrophils were determined. METHODS: A communication box was used for stress load on rats. SST-2 cells were inoculated in to rats in the tumor-inoculation groups. Two weeks after inoculation, the tumors were removed from the backs of the rats and weighed. The functions of neutrophils in the peripheral blood collected from the tail vein, were determined by the NBT deoxidization method. RESULTS: Tumor growth was enhanced when rats were loaded with either Ph or Ps stresses, but was inhibited when tumor cells were inoculated following either Ph or Ps stresses. These results show that chronological differences of loaded stresses influence immunological functions differently. The O2- production from the neutrophils stimulated by NBT-treated Staphilococcus aureus was suppressed in tumor-inoculated Ph and Ps groups, more markedly in the tumor-inoculated Ph group. It is logically relevant that the size of tumors increased in these groups, predominantly in the tumor-inoculated Ph group. On the other hand, O2- production from the neutrophils was enhanced and tumor growth decreased in tumor inoculated animals following either Ph or Ps stresses. CONCLUSIONS: Our experiments, it revealed that the function of neutrophils is strongly enhanced by stress load and O2- production is inhibited by the tumor inoculation as shown in stimulation tests. Therefore, our findings suggested that neutrophils may participate in the inhibition of tumor growth. PMID- 12638171 TI - [Regional differences in relationships between depressive symptoms and psychosocial factors among high school students: a comparison of Okinawa and Saga prefectures]. AB - OBJECTIVES: This study aims to examine regional differences in relationships between depressive symptoms and psychosocial factors among Japanese high school students in Okinawa and Saga prefectures in Japan. METHODS: The study sample was 3,234 students from 14 public high schools in Okinawa and 2,503 students from 12 public high schools in Saga. The students completed a self-administered questionnaire between September and November 2000. Depressive symptoms were measured using the Center for Epidemiologic Studies Depression Scale (CES-D). The psychosocial factors examined were participation in community events, life stress, self-esteem, perceived social support, and life environment. RESULTS: There was no difference in depressive symptoms by region. Students in Saga had higher levels of stress in the academic, teachers' and friends' domains compared to students in Okinawa. Students in Saga also evaluated their natural environment better than their counterparts in Okinawa. On the other hand, students in Okinawa were more likely to participate in community events than those in Saga. After controlling for sociodemographic variables, depressive symptoms were found to be significantly associated with life stress, self-esteem, and social support among students in both Okinawa and Saga. Only among students in Okinawa, however, were depressive symptoms significantly related to participation in community events. The psychosocial factors accounted for the same amount of variance in depressive symptoms for Okinawa and Saga students. Additionally, life stress in the teachers' domain had stronger associations with depressive symptoms among students in Saga than among those in Okinawa. The depressive symptom levels of Okinawa students seemed to be more strongly associated with self-esteem than those affecting their counterparts in the Saga prefecture. CONCLUSION: These findings show that there is similar pattern of associations between depressive symptoms and psychosocial factors among high school students in Okinawa and Saga, except for participation in community events. Life stress, self-esteem, and social support may be common factors associated with depressive symptoms among high school students regardless of region. Overall, the magnitude of the relationship between depressive symptoms and psychosocial factors was similar across Okinawa and Saga students. PMID- 12638172 TI - [An association (population attributable fraction) between smoking habit and mortality from all causes, cancer and lung cancer: NIPPON DATA80, 1980-1999. National Integrated Projects for Prospective Observation of Non-communicable Diseases and its Trend in the Aged]. AB - OBJECTIVES: The authors examined the relationship and the population attributable fraction (PAF) between smoking habit and death from all causes, including cancer and lung cancer. METHODS: The baseline data were collected in the National Cardiovascular Survey in 1980 carried out for all household members aged 30 years or older in 300 districts, which were randomly selected throughout Japan. The number of participants in the survey was 10,546. The vital status was ascertained in 1999, and he calculated the adjusted relative risk of mortality and the PAF of mortality attributable to stopping smoking. RESULTS: A total of 9,629 subjects were available for the final analyses. There were 165,190 person-years of follow up, and 2,011 deaths from all causes, including 579 deaths from cancer and 106 deaths from lung cancer. After adjustment for age and other cancer risk factors, smoking habit was associated with mortality from all causes, cancer and lung cancer. These associations had positive trends. The PAF (%) from all causes was 9.0 (male), 1.6 (female). The PAF (%) from cancer was 29.5 (male), 2.7 (female) and that from lung cancer was 42.9 (male), 7.1 (female). CONCLUSIONS: After adjustment for age and other cancer risk factors, smoking habit was associated with mortality from all causes, cancer and lung cancer. These associations had positive trends. The PAF (%) from all causes was 9.0 (male), 1.6 (female). The PAF (%) from cancer was 29.5 (male), 2.7 (female) and that from lung cancer was 42.9 (male), 7.1 (female). PMID- 12638173 TI - [Live birth distribution by time and place from 1981 to 1998 in Japan]. AB - OBJECTIVES: To investigate the diurnal rhythm of live births labored spontaneously, and the effects of obstetric intervention on birth time distributions. METHODS: The data of live births tabulated by time (one-hour intervals), date and birthplace throughout Japan between 1981 and 1998 were obtained with permission from the former Ministry of Health and Welfare. Together with an investigation of hourly birth numbers by place in each year, an annual transition of hourly birth rates in medical institutions and the diurnal rhythm of birth numbers in maternity homes and at home were analyzed using regression analysis. RESULTS: In every calendar year studied the hourly live birth numbers at hospitals showed a single-peak distribution pattern with maximum values at 13:00-15:00. The annual transition of hourly birth rates showed a 10% (birth numbers base) decrease in the 11:00-13:00 period in 1998 as compared with that in 1981, while there was a corresponding increase of 8% in the 13:00-15:00 period. Hourly birth numbers at clinics showed a double-peak distribution pattern with maximum values during the 11:00-12:00 and 14:00-15:00 periods in early 1980, while a single-peak distribution with a maximum value during the 13:00-15:00 period appeared in 1989 and has remained thereafter. Hourly birth rates (birth numbers base) increased by over 6% in the 13:00-15:00 and 17:00-20:00 periods over the past 18 years, while they decreased by 10% in the 9:00-13:00 period. The results at maternity homes were clearly different from those at hospitals and clinics. The live birth numbers totaled for the 18 years showed a double-phase distribution with a maximum value in the 6:00-7:00 period and a minimum value in the 19:00-20:00 period. The best-fit regression model for the obtained data was a sine curve with a maximum value at 6:00 (coefficient of determination 0.97). Hourly distributions of live births at home also fitted best to a since curve with the maximum value again at 6:00 (coefficient of determination 0.95). CONCLUSIONS: The results suggested that the timing of spontaneous live births follows a circadian rhythm and that obstetric intervention affects time distributions of live births by shifting over 10% of births during the night and early morning to a working day service time (9:00-17:00). PMID- 12638174 TI - [Role of the nectin-afadin system in organization of tight and adherens junctions in epithelial cells]. PMID- 12638175 TI - [Specific localization of ion- and water-channels and their functions in vectorial transports]. PMID- 12638176 TI - [Polarized expression of drug transporters and its physiological significance]. PMID- 12638177 TI - [Mechanisms of vesicle transport in polarized cells orchestrated by Rab GTPase]. PMID- 12638178 TI - [Molecular mechanism underlying cell polarity]. PMID- 12638179 TI - [Can genome sequences save tropical countries?: outcome form the genome sequences of malaria parasite, Plasmodium falciparum and malaria mosquito, Anopheles gambiae]. PMID- 12638180 TI - [Introduction to the practical analysis of DNA microarray data]. PMID- 12638181 TI - [Microbiological riskanalysis in food]. PMID- 12638182 TI - [Indoor air and human health--sick house syndrome and multiple chemical sensitivity]. AB - The number of complaints about the quality of indoor air has increased during the past two decades. These complaints have been frequent enough that the term "Sick House Syndrome or Sick Building Syndrome" and "Multiple Chemical Sensitivity" has been coined. Complaints are likely related to the increased use of synthetic organic materials in house, furnishing, and consumer products; and the buildings, furnishings, and consumer products; and the decreased ventilation for energy conservation in homes. Approximately thousand volatile chemicals have been identified in indoor air. The main sources of these chemicals are house materials, combustion fumes, cleaning compounds, and paints or stains. Exposure to high levels of these emissions and to others, coupled with the fact that most people spend more time indoors than outdoors, raises the possibility that the risk to human health from indoor air pollution may be potentially greater than the risk posed from outdoor pollutants. The complaints most frequently voiced with respect to Sick House Syndrome are irritations of the eye, nose, and throat; cough and hoarseness of voice; headache and mental fatigue. The syndrome of multiple chemical sensitivities is controversial subject with increasing impact on the field of indoor air quality. The controversy surrounding Multiple Chemical Sensitivity includes its definition, theories of etiology and pathogenesis, diagnostic, and life style. Multiple Chemical Sensitivity is considered the hypothesis that is a disease caused by exposure to many chemically distinct environmental substances at very low. PMID- 12638183 TI - [Toxicogenomics--a new paradigm of toxicology and birth of reverse toxicology]. AB - Toxicogenomics is, similar to other genome science fields, based on DNA microarray and/or DNA chip technologies, which has enabled a large number of gene expressions, thus, have been applied to such functional genomics as pharmacogenomics, oncogenomics, proteomics, metabonomics, etc. Toxicogenomics has advantages such as deducing the number of test animals and the test period, and adopts simpler techniques using new biomarkers. What toxicogenomics can offer to conventional toxicology is given herein. PMID- 12638184 TI - [Functional analysis of yeast homologue gene associated with human DNA helicase causative syndromes]. AB - Proteins having DNA helicase activity play very important roles in many processes involving DNA workings such as replication, repair, and recombination. In this decade, many DNA helicase genes have been cloned as the causative genes of human recessive heredity diseases. These are the causative genes for Xeroderma pigmentosum (XPB and XPD), Cockayne syndrome (CSB), diffuse collagen disease (Ku80), alpha-thalassmia (ATR-X), Bloom syndrome (BLM), Werner syndrome (WRN) and Rothmund-Thomson syndrome (RTS). The yeast homologue genes of these human DNA helicase genes exist. S. cerevisiae RAD25/SSL2, RAD3, RAD26, YKU80/HDF2 and RAD54 are the homologue for XPB/ERCC3, XPD/ERCC2, CSB/ERCC6, Ku80/XRCC5 and ATR-X/HX2, respectively. E coli. recQ gene and S. cerevisiae SGS1 are the homologue for all BLM, WRN and RTS. A search of whole genome of S. cerevisiae revealed that SGS1 is the sole homologue of recQ in S. cerevisiae. Thus it seems likely that SGS1 is a functional homologue of one or several human RecQ family genes. Many basic or essential functions are well conserved in the cells from lower eukaryotic to higher mammalian. The functional analysis in yeast could make an useful insight for the human homologue. To clarify the functions of S. cerevisiae Sgs1 and to get an insight into the functions of Blm, Wrn and Rts, in this study, we analyzed the phenotype of sgs1 disruptant and in detail the cause of the poor sporulation phenotype of sgs1 disruptants in relation to meiotic processes including meiotic recombination. The poor sporulation of sgs1 disruptants was complemented with a mutated SGS1 gene encoding a protein lacking DNA helicase activity; however, the mutated gene could suppress neither the sensitivity of sgs1 disruptants to methyl methanesulfonate (MMS) and hydroxyurea nor the mitotic hyperrecombination phenotype of sgs1 disruptants. The N-terminal 1-45 amino acid region and 698-1195 amino acid region of Sgs1, which including helicase domain and C-terminal RecQ conserved region with helicase activity, were required for complementation of MMS sensitivity and suppression of hyperrecombination of sgs1 disruptants in mitotic growth. The 126-400 and 596-1195 amino acid regions of Sgs1 were required for complementation of poor sporulation and of reduced meiotic functions. These regions required for the mitotic or meiotic functions of Sgs1 were well overlapped with the interaction regions of Top3 and Top2. Some of these results might explain the mechanism of the symptom of RecQ-related syndromes. PMID- 12638185 TI - Effects of gamma ray and electron-beam irradiations on survival of anaerobic and facultatively anaerobic bacteria. AB - An extension of the approval for food irradiation is desired due to the increase in the incidence of food poisoning in the world. One anaerobic (Clostridium perfringens) and four facultatively anaerobic (Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Enteritidis) bacteria irradiated with gamma ray or electron beam (E-beam) were tested in terms of survival on agar under packaging atmosphere. Using pouch pack, effects of two irradiations on survival of anaerobic and facultatively anaerobic bacteria were evaluated comparatively. E-beam irradiation was more effective than gamma ray irradiation in decreasing the D10 value of B. cereus at 4 degrees C, slightly more effective in that of E. coli O157, and similarly effective in that of the other three bacteria at 4 degrees C. The gamma irradiation of the bacteria without incubation at 4 degrees C before irradiation was more effective than that of the bacteria with incubation overnight at 4 degrees C before irradiation in decreasing the D10 values of these bacteria (B. cereus, E. coli O157, and L. monocytogenes). Furthermore, ground beef patties inoculated with bacteria were irradiated with 1 kGy by E-beam (5 MeV) at 4 degrees C. The inoculated bacteria in the 1-9 mm beef patties were killed by 1 kGy E-beam irradiation and some bacteria in more than 9 mm beef patties were not killed by the irradiation. PMID- 12638187 TI - Tropane alkaloids in auxin-independent root cultures of Physochlaina physaloides. AB - Adventitious and hairy root cultures of Physochlaina physaloides were established. These roots grew well and produced high amounts of tropane alkaloids (particularly hyoscyamine and 6 beta-hydroxyhyoscyamine) in auxin-free culture medium. The effects of basal media and temperature on the growth and alkaloid production of these roots were investigated. Both root cultures produced highest amount of tropane alkaloids in B5 medium though the optimum temperature for hairy roots were lower than that for adventitious roots. PMID- 12638186 TI - Tropane alkaloids in adventitious root cultures of Physochlaina physaloides. AB - Adventitious root cultures of Physochlaina physaloides were established and their growth and tropane alkaloid production were investigated. Although the roots cultured in Murashige-skoog liquid medium containing 1.0-3.0 mg/l IBA grew well, the addition of lower concentrations of auxins was more suitable for alkaloid production. The best alkaloid yield was obtained with 0.01 mg/l NAA in 1/2 MS liquid medium. In addition, the alkaloid differences between the cultured roots and the regenerated plant cultivated in the soil was studied. Cultured roots in this condition showed higher level of alkaloids when compared to leaf, stem and roots from the regenerated plant (3 months in field conditions). PMID- 12638188 TI - [Study on evaluating methods for the quality control of glycoprotein products. (IV)--erythropoietin products. Part 4]. AB - We have previously reported on peptide mapping of recombinant human erythropoietin (rhEPO) by liquid chromatography/mass spectrometry (LC/MS). Using this method, both peptides and glycosylation at each glycosylation site can be elucidated based on the mass chromatogram and mass spectrum. In this study, we evaluated the mass spectrometric peptide mapping with regard to its use in comparability assessment of both protein parts and carbohydrates parts in glycoprotein products. Models of closely related glycoprotein products used in this study are rhEPOs produced from three different sources. We demonstrated that the mass spectrometric peptide mapping can elucidate the identity of protein part, and differences in site-specific carbohydrates heterogeneity due to acetylation and sulfation among the three rhEPOs. Our method can thus be useful in comparability assessment of therapeutic glycoproteins. PMID- 12638189 TI - [Studies for analyzing the restricted ingredients such as 2-(2'-hydroxy-5' methylphenyl)-benzotriazole]. AB - 2-(2'-hydroxy-5'-methylphenyl)-benzotriazole (HMB) is nominated as the restricted ingredients in cosmetics in Japanese Pharmaceutical Affairs Act. So the analytical method for HMB was investigated by HPLC. After dissolving HMB in cream or oil with tetrahydrofuran and making the constant volume with methanol, the test solution was prepared. Twenty micro-liter of the test solution was determined by HPLC by using CAPCELL PAK C18 column, the mixture of methanol and water (17:3) as mobile phase, the detection wavelength at 306 nm. The concentrations of 10 to 80 micrograms/ml HMB gave the linear relationship between the concentrations and the peak areas. In creams and oil 0.1% and 1% HMB were sufficiently determined by the established method. PMID- 12638190 TI - [Estimated production by the official inspection of tar colors (including aluminum lakes) in fiscal year 2001]. AB - There were 162 official inspections of tar colors and their lakes in fiscal year 2001, and 162 samples were qualified. Total production amount of tar colors that passed inspection in Japan in fiscal year 2001 reached 134.8 tons. Tar color production amounts were described by month and by manufacturer. The food tar color produced in the largest amount was Food Yellow No. 4, accounting for 43.9% during this period. Their yearly total productions were decreased from 238.7 to 134.8 tons during 12 years. PMID- 12638191 TI - [Determination and survey of hexachlorobenzene in food red nos. 104 (phloxine) and 105 (rose bengale) by GC/MS]. AB - The contents of hexachlorobenzene (HCB) in Food Red Nos. 104 and 105 samples certified in fiscal years 1998-2001 were determined by GC/MS. All 32 samples contained HCB ranging from 0.2 to 32.9 mg/kg. The daily intakes of HCB based on the production of food colors was estimated to be from 0.018 to 1.456 ng, and their average (0.441 ng) corresponded to 0.0052% of the ratio for the Tolerable Daily Intake (8.5 micrograms for 50 kg body weight) of HCB developed by the IPCS. PMID- 12638192 TI - [Fluorometholone Reference Standard (control 011) of National Institute of Health Sciences]. AB - The raw material of fluorometholone was examined for the preparation of the "Fluorometholone Reference Standard (Control 011)". The analytical data obtained were: optical rotation, [alpha]20D = .5 degrees; UV spectrum, lambda max of 240 nm and specific absorbance in methanol at 240 nm = 350.7; IR spectrum, same as that of the Fluorometholone Reference Standard (Control 864); high-performance liquid chromatography (HPLC), total amount of impurities estimated to be less than 0.5%; loss on drying, 0.01%. Based on the above results, the raw material was authorized as the Fluorometholone Reference Standard (Control 011) of the National Institute of Health Sciences. PMID- 12638193 TI - [Tocopherol Succinate Reference Standard (Control 021) of National Institute of Health Sciences]. AB - The raw material of tocopherol succinate was tested for the preparation of the "Tocopherol Succinate Reference Standard (Control 021)". Analytical data obtained were as follows: infrared spectrum, same as that of the Tocopherol Succinate Reference Standard (Control 981); specific absorbance, E1% 1 cm (286 nm) = 40.7; thin-layer chromatography, no impurities were detected until 50.0 micrograms; high-performance liquid chromatography (HPLC), four impurities were detected and the amount of tocopherol succinate was estimated to be 99.0%; loss on drying, 0.11%. Based on the above results, the raw material was authorized as the Japanese Pharmacopoeia Reference Standard (Control 021). PMID- 12638194 TI - [Glycyrrhizinic Acid Reference Standard (Control 0211 and 0221) of National Institute of Health Sciences]. AB - The raw material of glycyrrhizinic acid was examined for preparation of the "Glycyrrhizinic Acid Reference Standard". The analytical data obtained were: UV spectrum: lambda max, 251 nm; and specific absorbance (E1% 1 cm) in ethanol at 251 nm, 149.6 (Control 0211) and 145.7 (Control 0221); IR spectrum, specific absorptions of raw material were consistent with that of Standard (Control 001). Also, thin-layer chromatography, no impurity was detected; high-performance liquid chromatography, several impurities were detected. The amount of each impurity was estimated at less than 0.2% and total amount of impurities was less than 0.6%. Based on the above results, the candidate materials were authorized as the Glycyrrhizinic Acid Reference Standard (Control 0211 and 0221) of the National Institute of Health Sciences. PMID- 12638195 TI - [Paeoniflorin Reference Standard (Control 011) of National Institute of Health Sciences]. AB - The raw material of paeoniflorin was examined for preparation of the "Paeoniflorin Reference Standard". The analytical data obtained were: UV spectrum: lambda max, 231.7 nm; and specific absorbance (E1% 1 cm) in methanol at 230 nm, 265.4; IR spectrum, specific absorptions of raw material were consistent with that of Standard (Control 985). High-performance liquid chromatography, several impurities were detected. The amount of each impurity was estimated at less than 0.1% and total amount of impurities was less than 0.2%. Based on the above results, the candidate material was authorized as the Paeoniflorin Reference Standard (Control 011) of the National Institute of Health Sciences. PMID- 12638196 TI - [The Endotoxin 10000 Reference Standard of the National Institute of Health Sciences (the Japanese Pharmacopoeia Endotoxin 10000 Reference Standard) (Control 0211)]. AB - To establish the fifth lot (Control 0211) of the Endotoxin 10000 Reference Standard of the National Institute of Health Sciences (the Japanese Pharmacopoeia Endotoxin 10000 Reference Standard), a candidate standard (CS) was prepared and then evaluated. The potency of the CS was assayed against USP Endotoxin Reference Standard (Lot G-1) and defined a s containing approximately 16,000 endotoxin units (EU) per vial by a collaborative study in which 5 laboratories participated. Based on the results, the CS was authorized to be the fifth lot of the Endotoxin 10000 Reference Standard containing 16,000 EU per vial. PMID- 12638198 TI - [The ninety year history of the Japanese Urological Association: striving for progress in urology as a specialty]. PMID- 12638197 TI - [The Endotoxin 100 Reference Standard of the National Institute of Health Sciences (the Japanese Pharmacopoeia Endotoxin 100 Reference Standard) (Control 0201)]. AB - To establish the third lot (Control 0201) of the Endotoxin 100 Reference Standard of the National Institute of Health Sciences (the Japanese Pharmacopoeia Endotoxin 100 Reference Standard), a candidate standard (CS) was prepared and then evaluated. The potency of the CS was assayed against USP Endotoxin Reference Standard (Lot G-1) and defined as containing approximately 130 endotoxin units (EU) per vial by a collaborative study in which 5 laboratories participated. Based on the results, the CS was authorized to be the third lot of the Endotoxin 100 Reference Standard containing 130 EU of endotoxin per vial. PMID- 12638199 TI - [Ureteral stricture after ESWL for ureteral calculi]. AB - BACKGROUND: There are many reports on the effects of ESWL, but few reports on the complications, especially remaining ureteral stricture after this treatment. Therefore we have retrospectively reviewed our cases to define the predisposing factors of this complication. METHODS: Since 1991 we have treated urolithiasis with ESWL using a Siemens Lithostar for the first therapy. We had 16 cases of ureteral stricture after this treatment. Ureteral stricture is the most common complication after ESWL treatment. To define the risk factor of the stricture we have compared 549 cases that were successfully treated between 1994 and 1996 without this complication. In these two groups we examined ages, sexes, chief complaints, size, position and components of the calculi, the degree of hydronephrosis, the frequency of ESWL, the presence of urinary tract infection, the duration of stone impaction and the after endourological treatment using multiple logistic regression analysis. RESULTS: Patients with the stone incidentally found and those with the UTI seemed to be more frequently associated with ureteral stricture, however there was not a significant difference. The hydronephrosis more than grade 3 (p = 0.025), the frequency of ESWL (p = 0.0325) and the after endourological treatment, especially TUL (p = 0.0184) were statistically significant among the other factors. The stricture occurred in 5 out of 29 patients with the hydronephrosis of grade 4 and 5 between 1994 and 1996. CONCLUSION: We should carefully treat patients with grade 3 or more hydronephrosis with ESWL. We should not repeatedly treat the patients with ESWL. We should take care of TUL treatment after ESWL. PMID- 12638200 TI - [Urolithiasis in those patients with myelodysplasia]. AB - OBJECTIVE: To evaluate incidence and treatments of urolithiasis in myelodysplastic patients. PATIENTS AND METHODS: During the past 27 years 303 myelodysplastic patients, 160 men and 143 women, have been treated and 50 of them were operated on with bladder augmentation procedures. Their medical records were retrospectively reviewed. RESULTS: Urolithiasis occurred 20 times in 15 patients, 11 men and 4 women, where calculi developed 18 times in the bladder and twice in the kidney. Prevalence of urolithiasis was 20% (10/50) in those who had undergone bladder augmentation and 2% (5/253) in those who had not had this surgery, where the difference was statistically significant between the two groups (p < 0.01). Overall, 5% of all the patients suffered from stones in the urinary tract. Bladder calculi were endoscopically treated in 11 occasions or were resolved with suprapubic lithotomy in 4 cases. Small stones in the bladder were spontaneously delivered 3 times and extracorporeal shock wave lithotripsy (ESWL) was necessary for a renal calculus in one patient. The last patient having a renal stone has been put under care. Majority of stone compositions consisted of magnesium ammonium phosphate (MAP). CONCLUSION: It was found that urolithiasis was 10 times more prevalent in those patients operated on with enterocystoplasty than those without bladder augmentation and that intermittent clean catheterization and regular bladder irrigation were of necessity to prevent urolithiasis for those having enterocystoplasty. PMID- 12638201 TI - [Laparoscopic pelvic lymph node dissection for staging procedure in patients with T1-T3 prostate cancer]. AB - PURPOSE: We evaluated the usefulness and morbidity of laparoscopic pelvic lymph node dissection (LPLND) as a staging procedure for prostate cancer. MATERIALS AND METHODS: Twenty-seven patients with T1-T3 prostate cancer scheduled for conformal radiation therapy underwent LPLND. RESULTS: The median operation time was 103 minutes (range; 58-137 minutes), and the median intraoperative estimated blood loss was 5 ml. (range; very little-273 ml.). This procedure covered obturator nodes and the median number of dissected lymph nodes was 8.0. Median days to oral intake and return to normal activity were 1.0 days and 1.0 days, respectively. CONCLUSIONS: LPLND appears to be a safe, minimally invasive and useful procedure as a means of accurate staging for patients with prostate cancer undergoing radiation therapy. PMID- 12638202 TI - [Acute normovolemic hemodilution in urologic surgery]. AB - PURPOSE: We report the clinical results and efficacy of acute normovolemic hemodilution (ANH) in urologic surgery. PATIENTS AND METHODS: Between October 1996 and February 2001 we performed ANH on 47 patients who were expected to have moderate blood loss during surgical procedures in our hospital. We then evaluated the postoperative hematological features and avoidance of homologous transfusion. RESULTS: Estimated median surgical blood loss was 400 ml (range 10-2,340 ml), and the median amount of whole blood collection was 800 ml (300-1,023 ml). In 14 patients whose blood loss was more than 1,000 ml, the hematocrit (Hct) level in the day after surgery was significantly higher than the Hct level calculated by blood loss. Ninety-eight percent of the series (46/47 cases) and ninety-four percent of patients (15/16 cases) who were underwent radical cystectomy and radical prostatectomy could avoid homologous transfusion. CONCLUSIONS: Our results indicate that ANH is useful during urological surgery, especially in patients with a blood loss of more than 1,000 ml during surgery. ANH is an efficient method for autologous transfusion by means of not only avoidance of homologous transfusion but also by saving red blood cells during surgeries. PMID- 12638203 TI - [Pseudotuberculous pyelonephritis associated with staghorn calculus: a case report]. AB - This report describes a 56-year old woman with an unusual form of chronic pyelonephritis mimicking tuberculosis on histopathological findings. She visited our hospital complaining of left flank pain. Left staghorn calculus and retroperitoneal abscess extending from the kidney were demonstrated on CT. No bacteria, including mycobacteria were identified in preoperative urine bacterial culture. Left nephrectomy with drainage of retroperitoneal abscess was performed. Microscopically, the nephrectomy specimen showed caseating granulomas, formed by epitheloid cells, highly suggestive renal tuberculosis. In spite of these findings, acid-fast bacteria were not revealed in the renal lesion nor the abscess, and cultures of the abscess for mycobacteria were also negative. Because of failure of identifying Mycobacterium tuberculosis, this case should be diagnosed as not renal tuberculosis, but pseudotuberculous pyelonehritis, which has been mentioned in recent literatures. Although this disease is not widely recognized, we must be aware of it to avoid unnecessary antituberculous therapy. PMID- 12638204 TI - [A case of urolithiasis associated with short bowel syndrome]. AB - We report a case of urolithiasis associated with short bowel syndrome. A 56-year old woman was admitted to our hospital for asymptomatic bilateral renal stones. She had received extensive resection of small intestine due to strangulating obstructive ileus 7 years ago (residual intestine, only 20 cm). Subsequently, she was in a state of short bowel syndrome. Plain film of kidney, uteter, bladder and computed tomography revealed bilateral renal stones (right 4 mm, left 10 mm). The left renal stone was successfully treated by extracorporeal shock wave lithotripsy. Since the right renal stone was small, no treatment was performed. The stone fragments were composed of calcium oxalate and calcium phosphate, and excessive urinary excretion of oxalate (103.8 mg/day) was observed. In this patient, urolithiasis was diagnosed to be due to enteric hyperoxaluria caused by short bowel syndrome. To prevent the recurrence of stone formation, she was treated with oral administration of calcium lactate, sodium/potassium citrate and magnesium oxide. We review the Japanese literatures on urolithiasis with short bowel syndrome. PMID- 12638205 TI - [Mechanism of action of bisphosphonate]. PMID- 12638206 TI - [Pathogenesis of osteoporosis--overview]. AB - Pathogenesis of osteoporosis was overviewed from standpoints of individual level, organ level and cellular level, respectively. As for the individual level, it is important to discriminate the pathogenetic factors related to low BMD or fractures. Independent from BMD, age, previous fractures, bone geometry, microstructure of the bone are related to fractures by influencing the bone quality. As for the organ level, pathogenesis of osteoporosis should be discussed from the view point of bone remodeling and its controlling factors. However, details of this process are still unclear. As for the cellular level, clarification of the roles of osteoblast, osteoclast and their interactions are essential in understanding the pathogenesis of osteoporosis. Recent discovery of Cbfa1, RANK/RANKL signaling and osteoprotegerin may open the new insight in the pathogenesis of osteoporosis. PMID- 12638207 TI - [Genetic factors of osteoporosis and polymorphisms]. AB - Osteoporosis is a common disease with a strong genetic factors. Twin studies have shown that genetic factors play an important role in regulating bone turnover and bone mineral density. These phenotypes are determined by the combined effects of several genes and environmental influences. A great deal of research has been done on candidate genes, among the best studied are vitamin D receptor gene. From a clinical standpoint, advances in knowledge about the prospect of developing genetic markers for the assessment of fracture risk will be used as targets for the design of new drugs for the prevention and treatment of osteoporosis. PMID- 12638208 TI - [Regulatory mechanism of bone resorption: roles of bone remodeling-regulatory cytokines 'osteokines' in osteoclast differentiation and function]. AB - The recent discovery of receptor activator of NF-kappa B ligand(RANKL)-RANK interaction confirms the well-known hypothesis that osteoblasts play an essential role in osteoclast differentiation. Osteoblasts express RANKL, a tumor necrosis factor alpha (TNF-alpha) family member, as a membrane-associated factor. Osteoclast precursors that express RANK, a receptor for RANKL, recognize RANKL through the cell-cell interaction and differentiate into osteoclasts. Recent Studies have shown that lipopolysaccharide and inflammatory cytokines such as TNF alpha and interleukin 1 directly regulate osteoclast differentiation and function through a mechanism independent of the RANKL-RANK interaction. Interferon-gamma and beta are also shown to be important negative regulators in osteoclastogenesis. These findings have opened new areas for exploring the regulatory mechanisms osteoclast differentiation and function by immune and inflammatory cells. PMID- 12638209 TI - [Possible mechanism of the specific action of bisphosphonates on osteoclasts]. AB - The mechanism of action of bisphosphonates on bone resorption was examined in mouse culture systems. Tiludronate did not inhibit the formation of osteoclasts induced by 1,25-dihydroxyvitamin D3 in cocultures of osteoblasts and bone marrow cells. Osteoclasts obtained from cocultures treated with tiludronate formed many resorption pits on dentine slices. However, pit formation by osteoclasts was dose dependently inhibited by tiludronate. Treatment of osteoclasts with risedronate disrupted actin rings, and inhibited pit formation by osteoclasts. Bafilomycin A1, a vacuolar H(+)-ATPase inhibitor, inhibited the pit-forming activity of osteoclasts but did not disrupt actin rings. Risedronate failed to disrupt actin rings in the presence of bafilomycin A1. These results suggest that acidification by vacuolar H(+)-ATPase is involved in specific incorporation of bisphosphonates into osteoclasts. PMID- 12638210 TI - [Effects of bisphosphonate on trabecular microstructure]. AB - Bisphosphonate is known to be deposited in resorption area. In our small animal study to evaluate the effect of bisphosphonate on trabecular microstructure, micro-CT demonstrated the different trabecular microstructure among the sham, ovariectomized (OVX) rats and the OVX rats which were treated with risedronate (0.1, 0.5, 2.5 mg/kg) or menatetrenone 30 mg/kg, or with the combination of risedronate 0.5 mg/kg and menatetrenone 30 mg/kg. The pattern of change on trabecular microstructure was also different between risedronate and menatetrenone; risedronate maintained trabecular connectivity, whereas menatetrenone thickened the trabeculae. The combination therapy with menatetrenone provided an additional improvement of trabecular microstructure to the single use of risedronate. PMID- 12638211 TI - [Development of bisphosphonates]. AB - Bisphosphonates (BPs), antiresorptive agents, have been established as first line drugs for treatment of osteoporosis and widely used all over the worlds. In Japan alendronate and risedronate, newer generation of BPs have been developed according to a evidence-based guideline and approved for clinical use in 2001 and in 2002, respectively. Although these BPs have been shown to have antifractures efficacy, great efforts have been made to explore convenient administration routes and schedules of BPs for individual patients. On the other hand, some of BPs are indicated for abnormal bone and calcium metabolism other than osteoporosis, including Paget's disease of bone, hypercalcemia associated with malignancy. Currently, extended indications of BPs are under development to the treatments of metastatic bone diseases, such as breast cancer, multiple myeloma, and rheumatoid arthritis. Since etidronate, first generation of BP also have the inhibitory effects on mineralization and hydroxyapatite crystal formation, application of this agent to ectopic ossification and calcification are very promising and under investigation. PMID- 12638212 TI - [Etidronate]. AB - Intermittent cyclical therapy with etidronate increases bone mineral density in spine and hip, and reduces the incidence of new vertebral fractures in women with postmenopausal osteoporosis, as well as steroid-induced osteoporosis. Seven years treatment with etidronate was reported to be safe, effective and well-tolerated. And additive effects of etidronate were documented in bone mineral density when hormone replacement therapy was combined. Recently many bisphosphonates have been commercially available in clinics. Although the potency of etidronate to inhibit bone resorption is relatively weak among those bisphosphonates, equal effects for others in osteoporosis can be obtained with the intermittent therapy, which is easy to be complied for the patients with less adverse events. In conclusion cyclical etidronate therapy is still important for osteoporosis therapy. PMID- 12638213 TI - [Clinical trials of alendronate]. AB - A number of large scale double-blind controlled studies with alendronate were carried out to evaluate therapeutic efficacy for osteoporotics. Alendronate administered orally at 5 mg per day for 7 years induced an average 8.2% increase of bone mineral density in the spine and 2.6% in the hip. Alendronate given orally for 4 years decreased significantly by 44% the occurrence of vertebral and non-vertebral fractures in patients with low bone mineral density and at least one fracture. In patients without any fracture, the results were significant for non-vertebral fractures when initial BMD was low. In histomorphometry with iliac bone biopsies, mineralization was normal, and trabecular bone turnover markedly decreased in patients receiving long-term dosing with alendronate. Alendronate is expected to have a potency to inhibit bone fracture occurrence in Japanese osteoporotics safely. PMID- 12638214 TI - [Risedronate]. AB - Risedronate (RSD) containing nitrogen, the third generation of bisphosphonate, has a powerful antiresorptive activity, without a stronger inhibition of mineralization. RSD is available for the treatment of osteoporosis. In Japanese the effective dose is 2.5 mg given orally daily, while in Caucasians it is 5 mg. This difference of dose suggests that RSD may be absorbed more efficiently in Japanese than in Caucasians. RSD administered orally at 2.5 mg per day for 48 weeks induced a 5% increase of bone mineral density in the lumbar spines in osteoporotic women. RSD, 5 mg daily, decreases vertebral and non-vertebral, especially hip fractures in elderly osteoporotic women. Thus, RSD is an important agent for the treatment of osteoporosis. PMID- 12638215 TI - [Methodology of bone mass measurement]. AB - Over the last several decades, methodologies for the assessment of bone mineral density have considerably progressed and at present bone densitometry has become a prerequisite for diagnosis, monitoring, and epidemiological studies of osteoporosis. However, until recently it was difficult to monitor the response to therapy by bone mineral measurement since the change of bone mineral density by therapy was small compared with measurement errors. On the other hand, bisphosphonate such as risedronate and alendronate have a strong effect on bone mineral density and bone densitometry would be expected to be used more widely in the evaluation of their therapeutic effect. In this paper, bone mass measurement clinically used at present is outlined. PMID- 12638216 TI - [Clinical practice guidelines for diagnosis and treatment of osteoporosis]. AB - The Japanese Society for Bone and Mineral Research organized a committee to make a new criteria of diagnosis for osteoporosis in 2000. In the criteria, cases who show atraumatic fractures in demineralized bone are made a diagnose of osteoporosis. In the cases who show no atraumatic fractures, patients whose bone mineral densities are less than 70% of mean density of young adult persons or whose X-ray films indicate coarse or unclear longitudinal trabeculae in vertebral body, are also made a diagnose of osteoporosis. A guideline for treatment of osteoporosis had been present in 1998, and another committee make ready for a new guideline in 2002. The new guideline is presenting as (1) increase in oral calcium intake and inpact exercise, prevention of fall, and wearing of external hip protector are recommended in addition to pharmacological agents, (2) in pharmacological agents, estrogens, calcitonins and bisphosphonates are effective to the patients with rapid bone loss, and (3) vitamin D3 derivatives and vitamin K2 are recommended for the patients with diminished bone formation. PMID- 12638217 TI - [Effective utilization of metabolic markers of bone]. AB - Although long-term treatment is essential to reduce osteoporotic fracture, the treatment is often interrupted, because patients or even physicians cannot recognize the effects of the treatment. The less power of the drugs to increase BMD, low precision of BMD measurement, and/or deformity of lumbar spine are responsible for the low recognition. Bisphosphonate and estrogen replacement therapy reduce the resorption of bone, which can be recognized by the measurement of resorption markers, such as NTX or deoxypyridinoline. Measuring these markers before and after the treatment, patients and physicians can recognize the effect within 1-3 months, which magnitude is usually more than the least significant levels. Confirmation of the effectiveness of the treatment through these markers may reduce the dropout rate of the treatment. PMID- 12638218 TI - [Treatment of osteoporosis provided from the orthopaedic surgeon's standpoint]. AB - Osteoporosis is associated with increased risk of fractures at most skeletal sites. Hip fractures have much greater prognostic significance in terms of health than any other single type of fracture. However, symptomatic vertebral fractures and other non-hip fractures also represent enormous morbidity and economic burdens, and signal increased risk of future fractures of all types, including the hip. There is convincing evidence that two bisphosphonates(alendronate and risedronate) reduce the risk of both spine and non-spine fractures. The objective of this paper is to summarize the literature concerning the consequences of osteoporotic fractures, and the effectiveness of pharmacologic agents for preventing fractures and their consequences, emphasizing a systematic, evidence based summary of treatment results from randomized, controlled trials that were published previously. PMID- 12638219 TI - [Role of physician on the treatment of osteoporosis]. AB - Physicians play an important role on the chronic management of the elderly patients. Those patients usually have multiple disorders and these situations lead to us to determine the priority of the treatments because of socio economical reason or tolerability of the patient against multiple drugs usage. Among the many involutional diseases, osteoporosis is considered to have less priority than the other diseases such as atherosclerosis, by physicians. However, fractures, most serious complication of osteoporosis, result in deteriorated ADL or QOL in osteoporotic patients. Therefore, when the elderly patients with osteoporosis are expected to have longevity or to have higher risks for future fracture, we should give a higher priority for the treatment of fracture prevention. Recent progress in the prevention of fractures allows us to treat osteoporosis with higher priority since the recently launched drug for osteoporosis such as bisphosphonates showed potent ability to reduce incident fractures. PMID- 12638220 TI - [Osteoporosis treatment by gynecologist]. AB - Postmenopausal bone loss is accelerated since women experience menstrual irregularity. Postmenopausal women lose their bone mineral density by 20 to 25% during 10 years after menopause, therefore early detection of risks for postmenopausal osteoporosis is mandatory for prevention of the disease. Because estrogen deficiency is the primary cause of postmenopausal bone loss, hormone replacement therapy can be a reasonable choice for the first treatment of osteoporosis. However, to those who have contraindications against estrogen or who complain severe estrogen-related symptoms, other medication using SERM and bisphosphonate should be considered. PMID- 12638221 TI - [Instruction on the use of drugs to an osteoporosis curative medicine]. AB - Patient compliance with drug therapy is an important factor in achieving successful therapeutic outcomes. Noncompliance is one of the major problems to be solved in the drug therapy of patients with chronic disease such as congestive heart failure, hypertension, and osteoporosis. Recently, it has been demonstrated that understanding of medication used to treat patients and illness, experience of adverse drug reactions, and complex route of drug administration are significant influencing factors on the incidence of noncompliance. Thus, to avoid the incidence of noncompliance and to complete drug therapy, we have to give a sufficient explanation about their medication and illness to the patients, especially patients receiving long-term therapy. PMID- 12638222 TI - [Glucocorticoid-induced osteoporosis]. AB - Glucocorticoid-induced osteoporosis is one of the major complications of long term exposure to supraphysiological doses of glucocorticoid. It has been recognized that bone loss is rapid, particularly in the first 6 months of the therapy. The skeletal effects are both dose and duration dependent; daily glucocorticoid therapy at doses of 7.5 mg/day of prednisolone or above leads to decrease bone mass and increase risk of fractures. The mechanisms which glucocorticoid induces osteoporosis are suppression of bone formation and increase of bone resorption. Hypogonadism also contributes to this pathological condition via direct suppression of sex steroids and indirect suppression through decreased secretion of pituitary hormones. Estrogen, vitamin D and its active analogues, and calcitonin, have been therefore used to prevent glucocorticoid induced osteoporosis; however, the effectiveness is somehow limited. Treatment with newly developed anti-resorptive amino-containing bisphosphonates such as alendronate and risedronate, showed significant increase of bone mineral density for both the prevention and the treatment of glucocorticoid-induced osteoporosis, as well as risk reduction of fractures in the patients. These bisphosphonates provide a better consequence for the treatment of glucocorticoid-induced osteoporosis. PMID- 12638223 TI - [Bone loss in type 2 diabetes mellitus--diabetic osteopenia]. AB - Loss of bone mineral content has been recognized as one of the chronic complications of type 2 diabetes mellitus, although its mechanism is not fully documented. A negative calcium balance due to both enhanced urinary calcium excretion and decreased intestinal calcium absorption has been occurred because of alteration of vitamin D metabolism and/or decreased parathyroid function. From the view point of bone cell metabolism, osteoblastic bone formation is suppressed by alternation of vitamin D metabolism, hypoparathyroidism, chronic hyperglycemia and insufficient insulin action. On the other hand, osteoclastic bone resorption is rather enhanced. The functional bone uncoupling system between osteoblast and osteoclast in type 2 diabetes mellitus could result in loss of bone density. PMID- 12638224 TI - [Osteoporosis associated with rheumatoid arthritis]. AB - Patients with rheumatoid arthritis(RA) develop both generalized and periarticular osteoporosis. Both of them are believed to be associated with increased production of inflammatory cytokines(TNF alpha, IL-1 beta, IL-6) and increased formation and activation of osteoclasts. Whether glucocorticoids work positively or negatively on generalized/periarticular osteoporosis is still controversial. RANKL has been shown to be expressed on T cells and fibroblast-like cells in the synovium, thus 'RANKL-RANK' pathway is likely to play an important role in periarticular osteoporosis and bone erosion as well as generalized osteoporosis. Among various therapies for generalized/periarticular osteoporosis in RA, anti cytokine antibodies/antagonists and osteoclast inhibitors including bisphosphonates are promising. PMID- 12638225 TI - [Osteoporosis associated with primary hyperparathyroidism]. AB - Primary hyperparathyroidism (pHPT) is cited as one of diseases which cause secondary osteoporosis and predominantly affects cortical bone. This disease occurs frequently during the postmenopausal period. Parathyroidectomy (PTX) is the only option available for the radical cure of pHPT and brings about marked increase in bone mass even in postmenopausal women. On the other hand, this disease is now recognized as a disorder with few overt manifestations, presenting as asymptomatic hypercalcemia. Even in these cases, a considerable increase in bone mass can be obtained by PTX. Bisphosphonate as well as estrogen and raloxifene are also reported to increase bone mass in elderly pHPT. patients. These drugs are considered as alternative therapy for mild pHPT patients who have many complications and/or are unfit for surgery. PMID- 12638226 TI - [Osteoporosis as a lifestyle-related disease]. AB - Although genetic factors determine the limits of peak bone mass, environmental factors can modify the outcome. Relation between lifestyle and osteoporosis is discussed, in terms of nutrition and habits. Significant link between calcium intake and bone mass has been reported. Although recommended daily allowance of calcium is 600 mg/day for adults, 850 mg/day or more shall be recommended later in life. Vitamin D insufficiency may lead to secondary hyperparathyroidism in the elderly, the condition that facilitates bone loss. Other nutrients that affect bone turnover include vitamin K, vitamin C, protein, potassium, salt, magnesium and phosphorus. Too much intake of caffeine or alcohol, as well as smoking is a risk factor of osteoporosis. Mechanical loading on the skeleton increases bone mass, therefore weight-bearing activity is recommended to gain or preserve bone mass. PMID- 12638227 TI - [Bone loss induced by GnRHa treatment in women]. AB - The hypoestrogenic state induced by gonadotropin-releasing hormone agonist(GnRHa) has been shown to be effective in the treatment of uterine leiomyoma and endometriosis but to induce bone loss. The BMD significantly decreased from baseline(-4.9 +/- 2.5%, mean +/- SD) after 24 weeks of treatment of leuprolide acetate depot(p < 0.01), and remained significantly below the baseline(-3.4 +/- 2.7%, p < 0.01) at 12 months after the treatment period. To minimize bone loss without compromising efficacy, several investigators have sought to 'add-back' sex-steroid hormones or other bone-sparing agents. Unresolved issues from these studies include ideal regimens and whether the add-back therapy prevents bone loss without compromising efficacy. GnRHa plus oral bisphosphonate therapy prevents bone loss without deteriorating the therapeutic effect of GnRHa. PMID- 12638228 TI - [Health examination of bone by the local government]. PMID- 12638229 TI - [Cooperation system in the treatment of primary osteoporosis]. AB - About 10 years ago, the selection of people with small bone mineral density has begun in each area of Japan. Even if there is a human with small bone density, it is not often possible to carry out the appropriate consultation in the medical institution in the region. It originates the bad cooperation between medical institution and bone measuring organization. In this country, osteoporotic patients are treated with various methods in the department of orthopaedics, medicine and gynaecology. As a result, they are not satisfied with the treatment. Efficiently, cooperation system are necessary in order to treat the human with small bone density. PMID- 12638230 TI - Characterization of the testis in congenitally ubiquitin carboxy-terminal hydrolase-1 (Uch-L1) defective (gad) mice. AB - The gracile axonal dystrophy (gad) mice are known to have a deletion within the gene encoding ubiquitin carboxy-terminal hydrolase-1 (Uch-L1) and show hereditary sensory deterioration and motor paresis. Expression of Uch-L1 is reported to be almost limited to the nervous system and testis. To understand whether Uch-L1, one of the major ubiquitin carboxy-terminal hydrolase (UCH) isozymes in the testis, affects spermatogenesis and other UCH isozymes (Uch-L3, L4 and L5) expression in the testis, we compared the testis between gad, hetero and wild type mice by histological, immunohistochemical analyses and RT-PCR. Histological analysis in 25-week-old gad mice showed shrinking of seminiferous tubules, decreasing total number of cells and enlargement of remaining cells in seminiferous tubules. By immunohistochemistry, a significant decrease (p < 0.05) in the number of proliferating cell nuclear antigen (PCNA) positive cells was observed. Expression of other UCH isozyme mRNAs was not apparently affected by Uch-L1 deficiency in 25-week-old gad mice. This study is the first report on the testis of gad mutant mouse. PMID- 12638231 TI - Preference of guinea pigs for bedding materials: wood shavings versus paper cutting sheet. AB - The preference of guinea pigs for bedding materials, wood shavings (WS) or paper cutting sheets (PS), was studied. Animals aged 8 weeks and 20 weeks showed a similar behaviour pattern during 30 min in the light, preferring WS to PS regardless of ages. Over both light and dark periods for 24 h, guinea pigs apparently preferred WS in the light, spending much more time resting in them than in PS. In the dark, the border-crossing was significantly more frequent than in the light, and the staying time was rather longer in PS than WS. The results suggest that guinea pigs prefer different bedding materials under light and dark conditions. PMID- 12638232 TI - Age-related changes on marking, marking-like behavior and the scent gland in adult Mongolian gerbils (Meriones unguiculatus). AB - Marking behavior, marking-like behavior [3], and changes of the scent glands were observed in aged Mongolian gerbils. In Experiment 1, changes in the marking and marking-like behavior with aging were evaluated in adult male and female Mongolian gerbils of an inbred strain aged 6 to 36 months. The frequency of marking behavior in males was significantly higher than females throughout the observation period except at 36 months of age. On the other hand, frequency of marking-like behavior in males, but not in females decreased with aging, significantly. In Experiment 2, changes of the scent gland in adult males and females aged 6 to 36 months were morphologically evaluated. Macroscopic examination revealed an increase in the size length and width of the glands of males aged 12 months and females aged 6 months. Histologically the glands of all the males and females aged 6 months developed moderately or well. Some of the 12 month-old males and females showed acinar atrophy of the glands, and all the females aged 18 months or more had highly atrophied scent glands. From these results, we concluded that there is no relationship between the changes of marking behavior and those of the scent glands in aged male Mongolian gerbils, and assume that marking behavior in aged animals does not have an important meaning as marking. In Experiment 3, marking and marking-like behavior in castrated adult Mongolian gerbils aged 16 weeks were observed. The result showed that marking behavior, not marking-like behavior was inhibited after castration. From these findings, we consider that generally marking behavior in Mongolian gerbils consists of androgen-dependent marking behavior and androgen-independent marking behavior (marking-like behavior). PMID- 12638233 TI - Establishment of an enzyme-linked immunosorbent assay for detection of hantavirus antibody of rats using a recombinant of nucleocapsid protein expressed in Escherichia coli. AB - A recombinant nucleocapsid protein of Hantaan virus (HTN) 76-118 strain expressed in E. coli was applied as a serodiagnostic antigen in an enzyme-linked immunosorbent assay (rHTN-ELISA) for detection of hantavirus antibody in rat sera. The sensitivity and specificity of the rHTN-ELISA were compared with those of the indirect immunofluoresent assay (IFA) using virus-infected cells. The sensitivity of rHTN-ELISA was similar to that of the IFA both in experimentally SR-11 infected rat and naturally infected rat sera. Sera showing a low antibody titer in IFA and suspected to be negative by other methods were also found to be negative in rHTN-ELISA. These results indicate that rHTN-ELISA is effective as a screening method for serodiagnosis of hantaviruses, because of its high sensitivity, specificity, safety and suitability for processing large number of samples. PMID- 12638234 TI - A new beige mutant rat ACI/N-Lystbg-Kyo. AB - A new beige-like coat color mutant was identified in the ACI/N rat colony. Other features characteristic of beige mutants, such as giant granule cells in various tissues, and prolonged bleeding time were also observed. The genetic complementation test, mating beige-like mutant with the authentic beige mutant rat, DA/Ham-Lystbg, revealed that the mutant gene is allelic to Lystbg. The new beige mutant allele was denoted Lystbg-Kyo. Molecular genetic analysis revealed deletion of exons 28, 29, and 30 of the Lyst gene owing to recombination between L1 elements in the mutant rats. Although the deletion was similar to that identified in DA/Ham-Lystbg rats, the putative deletion break points in L1 elements were different in the two strains. Further characterization of the ACI/N Lystbg-Kyo rats should make it useful as an animal model for human Chediak Higashi syndrome. PMID- 12638235 TI - Quantitative trait locus analysis of serum insulin, triglyceride, total cholesterol and phospholipid levels in the (SM/J x A/J)F2 mice. AB - Quantitative trait locus (QTL) analysis of serum insulin, triglyceride, total cholesterol and phospholipid levels at 10 weeks of age was performed in 321 F2 offspring from SM/J and A/J mice. Interval mapping revealed a total of 22 suggestive QTLs affecting the four traits: two insulin QTLs on Chromosomes (Chrs) 6 and 8; six triglyceride QTLs on Chrs 4, 8, 9, 11, 12 and 19; six total cholesterol QTLs on Chrs 1, 3, 4, 14, 17 and 19; and eight phospholipid QTLs on Chrs 2, 3, 4, 6, 8, 10 and 19. Gender influenced the expression of eight of the suggestive QTLs. The total-cholesterol QTLs on Chrs 4, 14 and 17, the triglyceride QTL on Chr 9 and the phospholipid QTL on Chr 4 were specific to females. The phospholipid QTLs on Chrs 2 and 6 and the insulin QTL on Chr 8 were specific to males. In addition, common QTLs involved in the regulation of some of the traits were identified. The female-specific QTL on Chr 4 appeared to be involved in the regulation of total cholesterol and phospholipid levels. The QTL on Chr 8 affected insulin and phospholipid levels, whereas the Chr 19 QTL was common to the three lipid parameters. PMID- 12638236 TI - D-galactosamine induced hepatocyte apoptosis is inhibited in vivo and in cell culture by a calcium calmodulin antagonist, chlorpromazine, and a calcium channel blocker, verapamil. AB - Studies were conducted in C57BL/6N Crj male mice and in cultured hepatocytes to clarify the relationship between galactosamine (GaIN) induced apoptosis and [Ca2+]i kinetics. Chlorpromazine (CPZ), a Ca(2+)-calmodulin antagonist, and verapamil (VR), a Ca(2+)-channel blocker each inhibited GaIN-induced DNA fragmentation and the appearance of apoptotic bodies. The kinetics of calcium uptake were evaluated using a calcium analyzer with the acetoxymethyl ester of fura-PE3 (fura-PE3/AM, 2.5 microM) as the calcium reporter. An increase in [Ca2+]i was detected in the cultured hepatocytes within 3 hours after treatment with 20 mM GaIN; this increase was inhibited by pretreatment with either 20 microM CPZ or 30 microM VR. Ca2+ imaging by confocal laser scanning microscopy showed that increase in [Ca2+]i after treatment with GaIN was initially localized around nuclei, while [Ca2+]i signals were later diffuse and observed throughout the cytoplasm. The activities of lactate dehydrogenase (LDH) and serum glutamate pyruvate transaminase (sGPT), used as indicators of plasma membrane damage and leakage, however, were not reduced by pretreatment with CPZ or VR. From these findings, we infer that the DNA fragmentation in GaIN-induced hepatocyte apoptosis is associated with an elevation in the perinuclear concentration of Ca2+, but GaIN-induced necrotic cell death is triggered through pathway(s) that are insensitive to blockage of Ca2+ influx and therefore appear to occur independently of elevation in [Ca2+]i. These results help to clarify the role of calcium flux in hepatocyte apoptosis and necrosis induced by exposure to hepatotoxins in vivo and in vitro. PMID- 12638237 TI - The Wakayama epileptic rat (WER), a new mutant exhibiting tonic-clonic seizures and absence-like seizures. AB - A new mutant, the Wakayama epileptic rat (WER), exhibiting both spontaneous absence-like behavior and tonic-clonic convulsions, was identified in a colony of Wistar rats. To determine clear seizure characteristics of this mutant strain, we analyzed the mode of inheritance of the convulsion and observed patterns of electroencephalogram (EEG) during the seizures. F1 progeny were produced between the founder male and normal females of the same colony. Animals were monitored through the inbreeding course to analyze genetic control of epileptic behavior. EEGs were recorded using affected animals in the F3-4 and post F13 generations. After the F2 generation, affected rats spontaneously exhibited both absence-like immobile behavior and tonic-clonic convulsions. The absence-like seizures were characterized by motor arrest and head droop. The tonic-clonic convulsions began with neck and forelimb clonus, wild jumping/running, and opisthotonic posturing, and evolved to tonic, then clonic convulsions. Most convulsion onsets occurred between 25-70 days of age. Mating experiments revealed that 0%(0/18) of the animals in F1, 10%(3/26) in F2, 17%(1/6) in backcross progeny and 86% (100/116) in progeny of crosses between epileptic rats showed tonic-clonic convulsions. Ictal cortical EEGs were characterized by 4-6 (5.1 +/- 0.4, mean +/- SD) Hz spike and-wave complexes in the absence-like seizures and by low-voltage fast waves in the tonic-clonic convulsions. This new mutant rat spontaneously exhibited both absence-like and tonic-clonic seizures. The tonic-clonic seizure was inherited as an autosomal recessive trait with 86% incidence. Thus, the new mutant rat may become a useful model for studying human inherited epilepsies. PMID- 12638238 TI - Developmental responses of two substrains of in vitro fertilized C57BL/6J mouse embryos to oxygen and amino acids. AB - The developmental response of in vitro fertilized embryos to oxygen and amino acids were compared between in-house bred C57BL/6JNrs (Nrs) and commercially available C57BL/6JSlc (Slc) mice. Under 20% oxygen, the percentage of embryos that developed to blastocysts and expanded blastocysts, and nuclear numbers were lower in Slc embryos than in Nrs embryos. Moreover, the nuclear number did not increase in Slc embryos during 72-96 h culture. Effects of amino acids were beneficial on development of Slc embryos under 20% oxygen, but inhibitory on blastocoel formation at 78 h under 5% oxygen. On the other hand effects of amino acids on Nrs embryos were observed in nuclear number at 72 and 78 h culture under 5% oxygen. Because the present study showed differences in sensitivity to culture conditions between the C57BL/6J substrains, care must be taken in embryo manipulation of this inbred strain in studies of embryo development or other studies. PMID- 12638239 TI - Experimental evaluation of cross-contamination between cryotubes containing mouse 2-cell embryos and murine pathogens in liquid nitrogen tanks. AB - It has been suspected that embryos stored in liquid nitrogen tanks may become contaminated with murine pathogens, if some pathogens had been introduced to the tanks accidentally. To examine this, we stored tubes containing embryos with tubes containing mouse hepatitis virus (MHV) or Pasteurella pneumotropica in liquid nitrogen tanks and examined whether progeny mice derived from the embryos were contaminated with the pathogens or not. After storing for 6 months or 1 year the frozen embryos were thawed and implanted into the oviducts of pseudopregnant female mice, and the mice were bred in vinyl isolators. We could not detect serum antibodies to MHV and isolate Pasteurella pneumotropica in the progeny mice, suggesting that cross-contamination between tubes in a liquid nitrogen tank scarcely occurs. PMID- 12638240 TI - Histology of vermiform appendix-like organ in slow loris. AB - The vermiform appendix-like organ (VALO) of the slow loris was investigated for its histology and immunohistochemical characteristics. The VALO has a much thinner wall with flat folded mucosa and shallower crypts than the cecal mucosa, while cellular components and population of the mucosa were similar to those of the cecum. No coalescent lymph nodules were seen in the submucosa. Immunohistochemically 5-HT-positive cells in the crypts and CD3- and CD8-positive lymphocytes in the lymph nodules were shown in the VALO as well as in the cecum. These findings suggest that the VALO is a low-differentiated vermiform appendix of the slow loris. PMID- 12638241 TI - Spontaneous development of dermatitis in DS-Nh mice under specific pathogen-free conditions. AB - Spontaneous development of dermatitis in DS-Nh mice under specific pathogen-free conditions was examined to verify the hypothesis [Exp. Anim. 46: 225-229, 1997] that Stapylococcus aureus (S. aureus) infection is causally associated with the dermatitis. Observation of the mice up to 28 weeks of age indicated that obvious dermatitis does occur under S. aureus-free conditions, though the incidence was low (six of 42 females and two of 90 males). Skin lesions in the absence of this bacterium showed histological changes very similar to those that can be observed under conventional conditions. In addition, hyperproduction of serum IgE was demonstrated in the dermatitis-positive mouse. These findings suggested that the dermatitis is triggered by IgE-mediated allergic reactions. PMID- 12638243 TI - Changes of heart rate during sexual behavior in the female rat. AB - The authors studied the physiological and behavioral responses during sexual behavior in female rats. For this purpose, electrocardiographs were recoded from conscious and unrestrained females using a radiotelemetry system. Heart rate during sexual behavior rose sharply following the introduction of a sexually active male, displayed a peak level of 425 bpm during male ejaculation, and then rapidly decreased. This pattern of heart rate in females before and after receiving ejaculation was similar to that in males before and after ejaculation. In the rate of decrease in heart rate, however, there was a significant difference between females and males. The present study is the first to show changes of heart rate during sexual behavior in female rats. PMID- 12638242 TI - Differences between BALB/c and C57BL/6 mice in mouse hepatitis virus replication in primary hepatocyte culture. AB - We previously showed that an intraperitoneal infection with mouse hepatitis virus (MHV) resulted in acute hepatic failure accompanying extremely elevated viral growth in the liver in interferon-gamma-deficient BALB/c (BALB-GKO), but not C57BL/6 (B6-GKO) mice. To examine the basis of the strain difference against MHV infection in interferon-gamma-deficient mice, viral replication in primary hepatocyte cultures from BALB/c and B6 mice with or without the IFN-gamma gene was compared in vitro. The MHV replication in BALB/c hepatocytes with or without the IFN-gamma gene was significantly higher than that in B6 hepatocytes with or without the IFN-gamma gene, suggesting that there is a strain difference in MHV replication in hepatocytes. Since a significant difference in MHV replication in hepatocytes was not observed between wild type and IFN-gamma-deficient mice of the same genetic background, the phenomenon is thought to be independent of IFN gamma. However, pretreatment of hepatocytes with recombinant mouse interferon gamma inhibited MHV replication in a dose-dependent fashion. The results are discussed with respect to the pathology of MHV infection in mice with or without the IFN-gamma gene. PMID- 12638244 TI - The rat lysosomal trafficking regulator (Lyst) gene is mapped on the telomeric region of chromosome 17. PMID- 12638245 TI - [Controversy about animal experimentation and animal rights]. PMID- 12638246 TI - [Model animals for muscular dystrophy]. PMID- 12638247 TI - [Drug design and animal experimentation]. PMID- 12638248 TI - [Life science and experimental animals]. PMID- 12638249 TI - [Safety of chemical substances used in animal experimentation]. PMID- 12638250 TI - [Experimental animals in the era of genomics]. PMID- 12638251 TI - [Legality of animal experimentation]. PMID- 12638253 TI - Animal experimentation in Europe: from its origins to its future. PMID- 12638254 TI - [The frequency of non-typeable Haemophilus influenzae systemic disease in children]. AB - 95 strains of Haemophilus influenzae (H. influenzae) isolated from blood of the patients with systemic infections were serotyped by staphylococcal coagglutination during the ten years from 1992 through 2001. As a result, 92 (96.8%) cases were caused by type b strains and 3 (3.2%) cases were caused by non typeable strains. Three cases with systemic infection due to non-typeable H. influenzae were reported. One patient was a premature neonate with sepsis and respiratory failure who had a fulminant course and died. The other two patients were a 3-year-old girl and a 1-month-old boy both with pneumonia. About their underlying conditions, one received intravenous steroid therapy and the other suffered from respiratory syncytial virus infection. They were treated with appropriate antibiotics and their clinical courses were satisfactory and uncomplicated. Non-typeable H. influenzae was isolated from not only blood but also the lower respiratory tract in all three cases. Systemic infection due to non-typeable strain is rare. But, it should be recognized as a substantial proportion of the serious infections caused by H. influenzae. PMID- 12638255 TI - [Isolation and serotypes of Vero toxin-producing Escherichia coli (VTEC) from pigeons and crows]. AB - To clarify the source and route of infection with Vero toxin-producing Escherichia coli (VTEC) in humans, we sampled gastrointestinal contents and isolated VTEC from wild birds captured to exterminate harmful birds between August 1997 and January 1998. Pigeons were caught in Sagamihara-shi and crows were caught in Sagamihara-shi, Kawasaki-shi, Yokohama-shi, and the Tokyo metropolitan area. The following results were obtained. 1) VTEC was isolated from 32 of 521 birds (6.1%) examined. Among pigeons, VTEC was isolated from 25 of 262 birds (9.5%) captured in Sagamihara-shi. Among crows, VTEC was isolated from 7 of 184 birds (3.8%) captured in Sagamihara-shi, but not isolated from any bird of 11.4, and 60 birds captured in Yokohama-shi, Kawasaki-shi, and the Tokyo metropolitan area, respectively. 2) Toxin was typed in 33 isolates. There were four VT1-producing isolates (6.5%), 27 VT2-producing isolates (88.7%), and two VT1, VT2-producing isolates (4.8%). 3) The serotypes of the isolates were: O78: H , 10; O152: H-, 7; O153: H19.2; O164: H-, 1; O128: H-, 1; O164/143: H-, and O1: HUT, 1. The serotype was unknown in 10 isolates. Among 10 isolates for which the serotype could not be determined, auto-aggregation was observed in one isolate. 4) EaeA was investigated in the 33 isolates, and 31 isolates (93.9%) possessed eaeA. The above findings showed that strains with same toxin types and serotypes of human diarrhea-derived VTEC were isolated from pigeons and crows, and the isolates frequently possessed eaeA, which is considered to have an important association with its pathology, suggesting that birds are involved in VTEC infection in humans as a source of infection. PMID- 12638256 TI - [Detection of new pigments related to purple urine bag syndrome]. AB - It is well known that participation of two kinds of pigments, indigo blue and indigo red, are concerned with purple urine bag syndrome (PUBS). However, there is no research which describes the participation of other pigments. We separated three new kinds of red pigments, other than indigo blue and indigo red, from four patients with PUBS. One of the pigments was not visible to the naked eye and appeared only when an ultraviolet ray was irradiated. The appearance patterns of indigo blue, indigo red and the three new red pigments were in complete agreement in two cases, but differed in the other two cases. Moreover, bacteria isolated from the urine were cultured in an indican-added alkaline liquid media (pH9), 80% or more bacteria produced purple pigments. In case 1, from deep-colored pigments, each bacterium produced both indigo blue and indigo red, although new pigments could not be observed in the in vitro media. PMID- 12638257 TI - [Possibility for the presence of Helicobacter pylori in drinking well water]. AB - Forty three well waters which are currently used as drinking water were studied for the presence of Helicobacter pylori. Using magnetic-beads purification and PCR amplification of H. pylori-specific gene, 4 of the 43 samples were positive for H. pylori-ureA gene (9.3%) and 1 of the 43 samples was positive for H. pylori 16SrRNA gene (2.3%). The presence of H. pylori-specific amplified product did not correlate with the type, depth and location of the wells. This study demonstrated that H. pylori can be transmitted via drinking water, especially well water in Japan. PMID- 12638258 TI - [10-yearly monitoring of prevalence of MRSA and antibiotic usage in a neonatal intensive care unit]. AB - From 1992 to 2001, we studied the prevalence of infants colonized MRSA and antibiotic usage for very-low-birth-weight infants in a neonatal intensive care unit at Asahikawa Kosei Hospital. We investigated nasopharyngeal swabs and stool samples every week after admission, and occasionally skin swabs, eye discharges and urines. Seventeen infants contracted nosocomial blood stream infection caused by MRSA which occurred between 1993 and 1996. Rate of infants colonized MRSA to all inpatients was 14.9% in 1992, which increased to 40.0% in 1994, and decreased to 7.1% in 2001. In each birth-weight group, rates of > or = 1,500 g, 1,000-1,499 g, and < 1,000 g infants were 6.5%, 45.0% and 60.0% in 1992, 24.5%, 100%, and 100% in 1994, 6.2%, 10.5%, and 21.4% in 2001. The longest period of antibiotic usage was 148.4 per 1000 patient-day in 1995 and decreased to 32.6 per 1,000 patient-day in 2001. The total value of antibiotics in 1995 was about yen 3,050,000, but in 2001 was about yen 470,000. PMID- 12638260 TI - [An imported case of falciparum malaria successfully treated with Artemether Lumefantrine in Japan]. AB - Spread of multi-drug resistant malaria in the endemic areas has made malaria control more difficult. Thus, WHO recommends combination therapy for the treatment of malaria. The aim of combination therapy is to improve efficacy and to reduce the incidence of resistance development to the each component of the combination. Particularly, the combination with artemisinin derivatives shows good outcome in Thailand where high resistance for mefloquine has already been found. We report the first case of falciparum malaria, successfully treated with Artemether-Lumefantrine in Japan. Artemether-Lumefantrine is a newly developed artemisinin-based combination agent for the treatment of uncomplicated multi-drug resistant malaria. This drug has proved highly effective and well tolerated by some clinical trials abroad. This Japanese female case showed a good clinical course without any side effect. PMID- 12638261 TI - [A case report of tuberculous pleuritis diagnosed by thoracoscopy using fiberoptic bronchoscope]. AB - A 27-year-old male visited the outpatient clinic of our hospital with the chief complaints of fever, right chest pain and shortness of breath. He was admitted to our hospital for detailed examination of the right hydrothorax. The pleural effusion obtained by thoracocentesis was exudative and negative for Mycobacterium tuberculosis. Since the titer of adenosine deaminase in the pleural effusion was abnormally high, antitubercular therapy was started under suspicion of tuberculous pleuritis. Thereafter, the patient's subjective symptoms and blood parameters improved. Necrotic tissues were obtained by pleural biopsy using the Cope needle. In order to make a definitive diagnosis, pleural biopsy was performed thoracoscopically. White tubercular lesions with a smooth surface were sparsely distributed on the pleura. Histopathologically, these lesions were characterized by central areas of caseous necrosis surrounded by epithelial cells and Langhans' giant cells. Therefore, they were considered to be granulomatous lesions. The patient was given a diagnosis of idiopathic tuberculous pleuritis, and was treated with four antitubercular drugs in combination. His clinical signs subsided, and he was discharged. This case indicates that the examination of the inside of the pleural cavity with a flexible bronchoscope, instead of thoracoscope, under local anesthesia is useful to diagnose patients having tuberculous pleuritis. PMID- 12638259 TI - Effect of amphotericin B dilution with various beverages on the survival of Candida albicans cells. AB - Amphotericin B (AMPH) has been generally used for prophylaxis or treatment of specific fungal diseases in immunocompromised patients. However, because it is difficult for children to ingest, mainly because of its bitter taste, it is often diluted with soft drinks. We therefore investigated the effect of dilution of AMPH with various beverages on its antifungal activity in vitro. Candida albicans cells were exposed for 30 min to AMPH diluted twofold with each of six commercially available beverages or distilled water, and percent survival was determined. The results showed 60% survival in the dilution with distilled water and higher survival when diluted with Yakult (136%; p < 0.01), orange juice (104%; p < 0.01), and coffee-milk (92%; p < 0.01). By contrast, lower survival was obtained when diluted with gum-syrup (54%), sweet cider (76%), and shaved-ice syrup (52%) with no significant differences from distilled water (60%), suggesting that these three beverages may be useful for diluting AMPH. The results of this study are a warning to medical workers that some methods of making AMPH more palatable considerably decrease its antifungal activity and may have a negative effect on host defenses against infectious diseases. PMID- 12638262 TI - [A case of imported Plasmodium vivax malaria from the demilitarized zone in South Korea]. PMID- 12638263 TI - [Professor doctor George Emil Palade at 90 years of age]. PMID- 12638264 TI - [Doubts on impact factor relevance in research evaluation]. PMID- 12638265 TI - [Intracellular events in T lymphocyte's activation]. AB - Adaptive immune responses are not initiated at the site where a pathogen first establishes a focus of infection. They occur in the organized peripheral lymphoid tissues, to which the pathogen or its products are transported, trapped, captured by specialized cells, called antigen-presenting cells (APC), which process and present the antigen to T lymphocytes. Activation of naive T cells requires two signals: the first signal is represented by the specific recognition of a foreign peptide fragment bound to a self MHC molecule, but this is not enough. The second signal, called co-stimulatory signal is represented by other molecules, expressed on the membrane or secreted by the APCs for which T cells express specific ligands. Binding of antigen to the T-cell receptor initiates a series of biochemical changes within the T cell, involving a large number of molecules. PMID- 12638266 TI - [New data about ITO cells]. AB - Ito cells (the fat-storing cells) are stellate cells located in the space of Disse. They can be selectively stained with gold chloride, but their distinguishing feature in routine histological preparations is the presence of multiple lipid droplets in their cytoplasm. Ito cells have the capacity to accumulate exogenously administered vitamin A as retinyl esters in lipid droplets. The quantity of vitamin A, which is stuck into Ito cells, decreases progressively in liver injury, like chronic alcohol intake. Also, the fat-storing cells are the principal cellular source of collagen and other extracellular substances in normal as well as fibrotic liver. Liver injury is followed by an increase in the number of the fat-storing cells. PMID- 12638267 TI - [Drug therapy at the beginning of the genomic era]. AB - The individualisation of drug therapy according to the genetic profile of each patient would allow to avoid the adverse effects and to reach the maximum therapeutic efficiency, therefore an optimum risk/efficiency ratio. This desideratum has become feasible in the genomic era by identifying and mapping a true mononucleotid polymorphism signature DNA (single nucleotide polymorphism fingerprint--SNP) and by using the new technologies. The present day data regarding the polymorphism of the genetic determinants involved in the response to drugs are synthetically shown, as well as the defining of the new fields- pharmacogenetics and pharmacogenomics. Although superposable and interchangeable up to a point, the term of pharmacogenetics rather refers to the study of the variability of response to drugs according to the genetic profile, the term of pharmacogenomics being reserved to the analysis of the genome (DNA and its products, RNA and proteins), in relation with the response to drugs. The differences among the individuals concerning the pharmacokinetics and the pharmacodynamics can be explained through the polymorphism of the substrata (enzymes, carrier proteins, receptors) that explains the genetic stratification of the population. The development strategies of the drug research and of the pharmaceutical industry will certainly be modulated by the new acquisitions in the field of pharmacogenetics and pharmacogenomics, with the inherent bioethical implications. New specific drugs for the patients possessing peculiar genotypes could be synthesized, the starting off of the prealable stratification of the patients according to their genotype. PMID- 12638268 TI - [DNA repair pathways and their involvement in human diseases]. AB - Integrity maintenance of the genome is crucial. Human DNA is vulnerable to damage arising from both endogenous and exogenous sources. Different DNA repair pathways counteract these potentially mutagenic accidents: damage reversal by methylguanine methyl transferase (MGMT), base nucleotide repair (BER), nucleotide excision repair (NER), mismatch repair (MMR) and repair of strand breaks. In some cases, DNA damage is not repaired but is instead bypassed by specialized DNA polymerases. The existence of human diseases associated with defects in DNA repair illustrates the importance of this process of quality control. Many of these human diseases have an increased susceptibility to cancer. PMID- 12638269 TI - [The beneficial effect of physical training in hypertension]. AB - Hypertension is present in epidemic proportion and is associated with a markedly increased risk of developing numerous cardiovascular disorders. All current treatment guidelines emphasise the role of nonpharmacological interventions, physical activity included, in the treatment of mild to moderate hypertension. A large number of studies have demonstrated that regular exercise reduces the incidence of hypertension. In addition to preventing hypertension, regular exercise has been found to lower blood pressure (10 mmHg average reduction in both systolic and diastolic pressure), improve lipoprotein-lipid profiles and insulin sensitivity. As part of the initial treatment, exercise is recommended for 12 months in patients with stage 1 hypertension, with no other coronary risk factors and no evidence of cardiovascular disease, and for as long as 6 months in those with other risk factor, but not diabetes. In patients with diabetes, cardiovascular disease or with stage 2 or 3 hypertension, drug therapy should be initiated first. Dynamic exercise of moderate intensity, 50-75% VO2max, (e.g. brisk walking, cycling) for 50-60 minutes, 3-5 times per week, is preferable to vigorous exercise because it appears to be more effective in lowering blood pressure. In addition to reducing hypertension, physical activity improves other cardiovascular risk factors. PMID- 12638270 TI - [Probiotics in gastrointestinal pathology]. AB - There are a variety of conditions in which the use of pro-biotics has proven effective:diarrhoea with its various causes, inflammatory bowel diseases, irritable bowel disease, colon cancer chemoprevention and hepato-portal encefalopathy. The pro-biotics have shown to be a promising therapeutical alternative for the future due to their lack of side effects and convenient mode of administration. Further, the potential of other pro-biotics is awaiting validation by clinical trials. PMID- 12638271 TI - [Microvesicular steatosis]. AB - BACKGROUND: "Bright" liver at ultrasonography predicts fatty liver. AIM: To assess prevalence and predictors of a bright liver. METHODS: Prevalence arm- Prospective collection of records of unselected patients undergoing liver ultrasound. Protocol arm--A sample of patients with bright liver underwent routine laboratory investigations, liver tests, HBsAg and anti-HCV. RESULTS: No difference was found in mean values of fasting plasma glucose, LDL cholesterol, total bilirubin, GOT, GPT, AP, GGT, and serum bile acids or in the prevalence of serum HBsAg and anti-HCV in bright liver vs control groups. Univariate analysis showed body mass index, age, total cholesterol triglycerides, albumin, HDL cholesterol to be significantly. CONCLUSIONS: Roughly 20% of patients undergoing US for routine clinical practice will have a bright liver and levels are the independent predictors of a bright liver. PMID- 12638272 TI - [The cell-cell interaction mechanisms and specific reactions in zona fasciculata cells from the adrenal gland]. AB - It seems that the particular intermediary disposal of zona fasciculata in the cortical ensemble of the adrenal gland is not insignificant, especially for the variability and for the glandular activities. Because of its specific cellular arrangement, displaying both positional and dimensional uniformity; it can be considered like a real islet of orderly cordonal symmetric tissue between two other cortical zones which present instead a very irregular cellular disposal. Distinct capsular fibrillar elements inserted around the cells and especially around small calibre vascular structures from zona fasciculata, are directly involved in the distribution of the secretory products. The concentration of the lipidic components is always more important at this cortical level than anywhere else in the whole adrenal gland, probably in connecting with the microsystem of channels from the entire surface of the zona fasciculata cells. PMID- 12638273 TI - [ Aspects of iatrogeny through communication in complex oral rehabilitation]. AB - Human intercommunication means transfer of information through many channels, with codes that have biological and socio-cultural determinism. The biological determinism is identifiable through the development of the phonetics and ergomotricity functions, through the possibility of perception of the sensorial organs. The channels of human intercommunication can be verbal, linguistic (the most specialised channel), and nonverbal, extralinguistic channels (mimics, written model, drawing, walking), that are effectively implicated in defining a direct dialogue between the dentist and the patient, completing and emphasising the communication. When the communication through the standard channels is disturbed, the dentist can't understand the patient's expectations, so the patient will not perceive the opportunity, the necessity and the efficiency of the complex oral rehabilitation. All these factors are contributing to the initiation of the iatrogeny through communication. PMID- 12638274 TI - [Ischemic left ventricular diastolic dysfunction at females: clinical aspect]. AB - The prevalence of the coronary heart disease at females becomes close to that of the male gender at older ages. However, studies of the assessment of ischemic cardiac disfunction include a few women. The objective of the paper is the analysis of the clinical profile of women with ischemic heart disfunction. METHODS: There have been studied 189 patients with coronary heart disease (chronic myocardial infarction, stable angina, ischemic cardiomyopathy): 63 females and 126 males. The prevalence of the coronary diseases, risk factors, symptoms, lipidic profile, radiologic and echocardiographic alterations were analysed. RESULTS: Mean age of the male group was 56.8 +/- 12.2, and of the female was 66.3 +/- 10.9 years, significantly older (p = 0.00001). Ischemic cardiomyopathy had a greater prevalence at women (33.3% vs 11.9%, p = 0.0004), while myocardial infarction dominated at the male gender (61.9% vs 28.6%, p = 0.0001); stable angina had similar rates (20.6%, respectively 19%). Episodes of cardiac decompensation were more often at women (20.6% vs 7.1%, p = 0.0003). Arterial hypertension prevailed at females (77.8% vs 60.3%, p = 0.01), and smoking at males (72.2% vs 7.5%, p = 0.00001). Women declared more often dyspnoea (p = 0.02) and the resting heart rate was higher (87 +/- 23/min vs 75 +/- 13/min, p = 0.0007). Mean values of cholesterol were higher at women, particularly LDL: 165 +/- 49 mg% vs 149 +/- 46 mg%, p = 0.04. At X-ray examination, cardiomegaly and pulmonary congestion were dominant at females. At echo examination, the ejection fraction had similar values (52.3 +/- 9.3% at women and 51.2 +/- 11.6% at men), but left ventricular hypertrophy and diastolic disfunction were prevalent at women (87.3% vs 69.1%, p = 0.01). CONCLUSIONS: Ischemic heart disfunction at females appears at older ages, with the dominance of ischemic cardiomyopathy and angina. At similar levels of sistolic disfunction, the diastolic disfunction, the left ventricular hypertrophy and the cardiac decompensations are more frequently at female gender. PMID- 12638275 TI - [The scintigraphic assessment of the patients suffering from cardia achalasia, treated by pneumatic dilatation]. AB - The esophageal scintigraphy is a technique used to evaluate the motility and the clearance of the esophagus. The patients with achalasia of the cardia were evaluated, before treating them with pneumatic dilatation and after. The reason is to evaluate the efficiency of the pneumatic dilatation technique in the treatment of esophageal achalasia. The pneumatic dilatation with an instrument type Rigiflex is a well/handled and used technique in our clinic. In the study 22 patients were included and we had in view the esophageal transit time and the residuary radioactivity before and after the pneumatic dilatation. We established an important statistical improvement of the data obtained after the dilatation comparative with the data before the treatment (p < 0.01). PMID- 12638276 TI - [The link between glomerulonephritis and hydatid disease ]. AB - The presence of glomerular damage was evaluated for 338 patients, adults (298) and children (40), with pulmonary or hepatic hydatid disease. A descriptive study was used. The glomerular syndrome was present in 2 children (5%) with hydatid disease, 22 adults (11%) with liver echinococcosis and 28 adults (18%) with pulmonary echinococcosis. An early stage diagnosis and proper treatment will determine the reversibility of glomerulonephritis. PMID- 12638277 TI - [The pathological mechanisms of glycoregulation disturbances in chronic hepatitis B and C]. AB - Despite the abundance of reports concerning the increased frequency of diabetes and impaired glucose tolerance in chronic liver diseases, the mechanisms underlying this phenomenon have not been resolved. 30 patients with hepatitis C virus (HCV) infection and 17 with hepatitis B virus (HBV) infection who showed an altered responds to a standard oral glucose tolerance test were investigated in order to evaluate their pancreatic-endocrine features. We have also evaluated 40 patients (20 with HCV infection and 20 with HCB infection) who developed diabetes after diagnosis of liver disease. Patients with HBV infection showed signs of enhanced insulin resistance but overt diabetes develops in those who, in addition to insulin insensitivity, have a relative defect of insulin secretion. In patients with HCV infection the significantly lower plasma insulin and C-peptide levels suggest that impairment of insulin secretion is the main mechanism leading to both glucose intolerance and overt diabetes. PMID- 12638278 TI - [Clinical benefits of montelukast sodium treatment in chronic adult asthma]. AB - This paper presents the results of Montelukast sodic therapy in 16 asthmatic patients, in a prospective study. Montelukast sodic was given in severe chronic asthma treated by inhaled or systemic steroids before. Patients were followed up for three months regarding: clinical symptoms, respiratory function (FEV), blood and sputum eozinophyls. Short term therapy showed a great improvement in day and night symptoms, a reduction of sputum and blood eozinophyls and an increased FEV. Long term therapy could be protective for asthma exacerbations and will provide constant good life quality of asthmatics. PMID- 12638279 TI - [Sequential esophageal scintigraphy contribution on cardia achalasia diagnosis]. AB - Achalasia is a mortality disorder described in 1674 by Willis. It has a small incidence in the general population. The main symptom is dysphagia. The golden standard for diagnosis is esophageal manometry. Esophageal scintigraphy is a non invasive method of exploration: the technique is simple, it lasts five minutes and is more sensitive than the radiologic exam. The esophageal scintigraphy was made in the Nuclear Medicine Laboratory of the Sf. Spiridon Hospital using a gamma scann Diacam--Siemens. The parameters measured with esophageal scintigraphy are: esophageal transit time, residuary radioactivity. Our study included a number of 37 patients with achalasia with ages between 16 and 80 years (the average--47, 3 years). Our conclusions are the same with the data from literature. PMID- 12638280 TI - [Functional digestive disorders and the relationship with psychiatric diseases]. AB - Functional gastrointestinal disorders are frequent causes for medical consultations. Features related to disease, patient or the environment could influence the medical referral. The aim of the study was to estimate the importance of psychiatric disorders in patients with functional gastrointestinal disorders. MATERIAL AND METHOD: The study was designed to have two phases. Phase I consisted in studying the prevalence of Irritable Bowel Syndrome (IBS) and psychiatric past history in a random sample of 338 from an urban community of 18000 people. Phase II consisted in studying the presence of psychiatric history in patients referred for IBS symptoms. RESULTS: The prevalence of IBS symptoms in the general population was: 14.4% (8.4% in males and 17.7% in females). Psychiatric positive history was present in 6.2% of the general population and in 6.1% of IBS subjects (p > 0.05). Psychiatric disorders were more common in the IBS subjects who were seeking medical assistance (18%) than in subjects with same symptoms from the general population (6.2%) (p < 0.05). CONCLUSIONS: Psychiatric disorders may influence the medial referral for patients with IBS. PMID- 12638281 TI - [Splenic hydatic cyst. Observations upon 38 cases of splenic echinococcosis]. AB - 15 patients with spleen hydatic cysts have been operated between 1986-1998, in percentage of 4.6% from the total number of hydatic cysts--8 females and 7 males with mean age of 44 years (17-70). They have been admitted for abdominal pain (66.6%) and at the clinical examination they presented a cystic tumor at the level of the spleen. In all the patients we performed splenectomy. In conclusion the hydatic cyst of the spleen is rare, but possible, the treatment being only surgical. PMID- 12638282 TI - [Lamotrigine in the treatment of pharmaco-resistant epilepsy in children]. AB - Lamotrigine (LTG) is an antiepileptic drug whose therapeutic effect may be due to inhibition of the excitatory transmitter glutamate release. The aim of this study is to evaluate LTG efficacity and tolerability in the therapy of drug-resistant epilepsy in children. LTG is tested in 27 patients with different forms of epilepsy. Results show a satisfactory response and the low adverse effects of LTG in agreement with literature data. The therapy also improve cognitive function and relationship. PMID- 12638283 TI - [The ecological accident on the Siret River. A possible cyanide intoxication of children]. AB - THE PURPOSE: Of our study is to assess the gravity of cyanide intoxication of children, in the ecological accident in January 2001. MATERIAL AND METHODS: We included in our study 127 children aged between 2 months and 17 years. They were hospitalized in January and February 2001, in the "Sf. Maria" Children's Hospital. Case analysis was based on data of the emergency unit and units where the children were sent after primary care. We evaluated the patients from clinical and paraclinical point of view and we took samples of biological products in order to confirm the diagnosis. RESULTS: Age distribution was 0-1 year (4%), 1-3 years (7.8%), 3-6 years (9.4%), 6-12 years (33.8%), and 12-18 years (42.5%). The lapse of time between the ingestion of contaminated fish and symptomatology was variable. Measures were taken: removal from toxic contact, hospitalization, antidote administration, and treatment of complications. DISCUSSIONS: Although literature data shows that this intoxication appears after the age of three, we found that mother ingestion of contaminated fish, determined intoxication in nursings. Symptoms appeared at 3-12 hours after fish ingestion from the contaminated river and it was suggestive for the diagnosis. First aid measurements were necessary in specialized emergency unit and antidote administration sometimes only in clinical suspicion of intoxication. Correlation between clinical findings and toxic concentrations in urine samples confirm the diagnosis. CONCLUSIONS: Our study tries to highlight those ecological accidents with cyanides present important consequences on people and animal's health. The therapeutic intervention must be fast in order to stop blood distribution of the toxic. PMID- 12638284 TI - [Systemic lupus erythematosus in children. I: clinical and age dependent evolutive characteristics]. AB - The individualized, retrospective study of 14 children with SLE (4-16 years) pointed out a series of clinical and age dependent evolutive characteristics. Below the age of 10 years old (lot 1:2 boys and 4 girls), SLE started as a prolonged fever syndrome (5-16 weeks) in the majority of cases; for 2 children the severe poliarthritis resistant to the AINS therapy is associated with the durable absence of the antinuclear seric antibody (ANA). For the same age group a high frequency of neurological manifestations (5/6 cases) was noticed. After the age of 10 years old (lot II: 8 girls) the symptoms incidence at debut is close to the one of the adult, the cutanat and renal manifestations in evolution were dominant (7/8 cases). The 7 months absence of ANA characterises a case that started with hepatomegaly, severe neurological and physiological manifestations and microscopic hematuria. The follow-up lasted until the age of 16 years old; the patients were clinically tested for severe renal complication. The correct application of the classical criteria of diagnostic (ARA, 1982), and in the last few years the application of the ponderat score (Mayer, 1998), allows us to establish an early diagnostic and a rapid evaluation of a relapse, therefore influencing the treatment. PMID- 12638285 TI - [Platelet adhesion and antioxidant therapy in experimental stress]. AB - We have investigated the platelet activity, by platelet adhesion index and the end products of lipid peroxidation--MDA. Platelet adhesion index was established using Rovatti's method. Determinations were carried out on experimental model consisting of five groups of white male Wistar rats. In the vitamin E supplemented group, the platelet adhesion index was constantly correlated with MDA level. Thus there is a correlation between the intensity of the oxidative stress and vitamin E supplementation. The imbalance between pro- and antioxdants in experimental stress may enhance the platelet adhesion via arachidonic acid metabolism and endoperoxides formation. Steady administration of vitamin E in the experimental stress, due to its antioxidant properties, causes the progressive decrease of peroxidation and of platelet adhesion. PMID- 12638286 TI - [Could biological aggressivity markers of breast cancer alter the therapeutic course? Preliminary results]. AB - The aim of this study is to justify an individual therapeutical attitude in breast cancer, related to diversity of breast tumors, aggressiveness grade and metastatic potential. Between January 2000--December 2001, 150 patients were admitted with breast cancer (stage II and III) and underwent surgery in our department. We selected 75 cases in our study. In 51 (68%) cases the first therapeutical method was surgery, in 15 (20%) cases surgery was performed after chemotherapy, in 2 (2.66%) cases after radiotherapy and after chemotherapy and radiotherapy in 7 (9.33%) cases. We evaluated several classical factors and new immunohistochemical markers with an important value for diagnosis, prognosis and therapy: oestrogen and progesterone receptors, c-erb B2, pS2 and p53 proteins, von Willebrand factor. Several factors had a predictive role regarding the response to chemotherapy. These predictive factors will improve the histopathological diagnosis. The oncoproteins and hormonal receptors also will evaluate with more accuracy the metastatic risk and will assure a better therapy decision. PMID- 12638287 TI - The multiple tumor syndrome. Personal experience. AB - The multiple tumor syndrome is an unusual pathologic condition, which consists in association of multiple malignancies in the same patient. Seven cases are discussed: two women, five men, aged 32-70 years. The period between the two neoplasias was 2-23 years (in 6 cases). In one case the two malignancies appeared concomitantly. The hematological malignancies were: multiple myeloma: 2 cases; chronic granulocytic leukemia: 2 cases; chronic lymphatic leukemia: 3 cases. In four cases, the solid tumor followed the hematological malignancy at variable periods (2 and 4 years). In other two cases, the solid tumors preceded the hematological malignancy with 2 years, 23 years respectively. The solid tumors were genital cancers, malignant melanoma, spino-cellular carcinoma, thyroid cancer, hemangiosarcoma. In a single case the second tumor was a hematological malignancy too (NHL-diffuse lymphocytic lymphoma). Possible implications of previous therapy and environmental factors are discussed. PMID- 12638288 TI - [Maternal immunophenotypic profile in normal pregnancy and preterm birth]. AB - During normal pregnancy, the maternal immune response achieves a split tolerance state towards the fetal allograft, expressed by immunophenotypic profile. This may be impaired in the preterm birth. Our study aimed to analyze the immunophenotypic profiles of the peripheral and retroplacental blood in women with normal pregnancy and preterm birth. Blood leukocyte populations were assessed by flowcytometry in nine normal at term pregnancies, five preterm births and eight nonpregnant controls. Our results showed in normal pregnant women as compared to controls, a significant decreased number of lymphocytes, monocytes and B cells, while granulocytes, T-lymphocytes and the T CD8+ subsets were elevated. The CD4/CD8 ratio was significantly decreased, especially in the retroplacental blood. The preterm birth group demonstrated a significantly elevated number of lymphocytes and B cells, lower T CD8+ cells and an increased CD4/CD8 ratio, in comparison with normal pregnancies group. CONCLUSION: Normal pregnancy determines a suppressive immunophenotypic profile that is impaired in the preterm birth suggesting its immunological involvement. PMID- 12638289 TI - [Hemophilus influenzae meningitis- the experience of infectious diseases department between 1984-2001]. AB - Our paper presents a clinico-biological and therapeutical study of the cases admitted in Infectious Diseases Hospital between January 1984-December 2001. We studied the records of all the patients that suffered from meningitis with Haemophilus influenzae. In above mentioned period 40 cases of Haemophilus influenzae meningitis was admitted; 6 patients (15%) died. A number of 13 cases was registered for the first five years, mainly in the childhood-32 cases (80%) for infants. Bacteriological diagnosis was made by bacterioscopia for 16 patients, bacteriological cultures-30 patients, latex test, performed in the last five years, in 8 cases. In patients who died, the bacteriologic culture was positive in 5 cases and bacterioscopia in 4 cases. The therapy was performed in resonance with antibiograme results for the involved strains. The Haemophilus influenzae meningitis is an acute and severe disease of the childhood, with an important risk of death, despite the rigorous therapeutical measures. PMID- 12638290 TI - [Leptospirosis-clinico-biological and therapeutical aspects-study of 256 cases]. AB - Leptospirosis is a zooantroponosis manifested as an infectious disease with a severe evolution, with liver and renal failure and hemorrhagic manifestation. The aim of this study was to present the clinico-biological investigation and the therapeutical aspects of 256 cases of leptospirosis admitted in the Infectious Diseases Hospital Iasi during 4 years (1998-2001). The diagnosis was based on the study of clinico-biological and therapeutical parameters (risk factors, fever, mialgia, headache, algic, eruptive, meningeal, hepatorenal, hemorrhagic syndromes with the serological evidence of the serotypes of leptospirosis); most of the cases were registered in 1998 (23% cases), in adults (31.2%) and male (75.22%). All the patients were serologic confirmed with leptospirosis (RAL and RFC), and L. icterohaemorhagiae was isolated most frequently. The standard anti leptospirosis treatment was administered with favorable evolution; 6 deaths were registered from the severe forms of the disease. The leptospirosis is an emergent disease with the possibility of severe evolution in some forms, with liver and renal failure. PMID- 12638292 TI - [A study regarding the anatomy of the main arterial pedicles of the greater omentum]. AB - Hepatic artera takes part to the greater omentum vascularization by the right gastroomental artera (most frequently as a branch of the gastroduodenal artera). The investigations on 100 anatomical specimens from adult subjects revealed a mean value of the artera length of 19.30 cm and a diameter of 1.5-3 mm. The lienal artera participates to the greater omentum vascularization by the left gastroomental artera (with a mean length of 10-18 cm and a diameter of 1.5-2.0 +/ 0.3 mm. The arterial arch of the large gastric curvature has a length of 45 +/- 5 cm; the functional anastomotic point is localized at 10-15 cm left size reported to the median line of the large curvature. PMID- 12638291 TI - Tolerance to penicillin in Staphylococcus pyogenes isolated from skin infections. AB - The tolerance to penicillin of S. pyogenes has been mentioned in pharyngeal strains and associated by some authors with penicillin treatment failure in streptococcal angina. In this study we followed the presence of tolerance in S. pyogenes strains isolated from children hospitalized in the Clinic of Infantile Surgery and Clinic of Dermatology Iasi for skin infections. From 138 samples of pus examined we isolated 14 strains of S. pyogenes. In 7 strains we pointed out the tolerance to penicillin by both, beta-lactamase disc test and time-killing curve methods. We did not correlate to our cases the tolerance presence with penicillin treatment failure. PMID- 12638293 TI - [The surgical importance of the thyro-laryngo-pharyngeal lymphatic interconnections]. AB - The paper presents a study of the evolutional morphogenetic stages of the lymphatic and conjunctive neck structures formation. The authors make particular appreciations concerning the pattern of organization and also, the functional connections of lymphatic and conjunctive elements at intravisceral, perivisceral and perivascular level. Our study proves that these structures form the barriers but also, the spread pathways of the tumoral and inflammatory process which, after an endocavitary development, diffuse either on the adjacent organs or to the main cervical lymph nodes. We emphasized the importance of these anatomical structures in the neck oncological surgery. PMID- 12638294 TI - [Family involvement in dental health education of school children]. AB - Education for oral-dental health in children is that component of general health education aimed at creating cultural health models, cultivating in the young generation a healthy hygienic behaviour and outlying the opinions about the ways dental disorders can be prevented and treated. The most important goal of health education is to contribute to the preservation/improvement of children's oral health status. GOALS: This study has two main goals: to assess the exact health education knowledge of the questioned parents and to evaluate their involvement in the oral health education and promotion. MATERIAL AND METHOD: This study included 95 parents, aged between 25 and 49 years, with children in primary schools. For data collection a questionnaire was used. The questions were grouped on common features: food habits and healthy diet, causes of oral disease, prevention of oral disease, dental visit habits, oral hygiene habits. RESULTS: The study revealed that parents have a moderate knowledge about dental health education and dental caries prevention, no significant sex differences being found, and poor knowledge about periodontal diseases prevention. As to food hygiene, parents proved a sound knowledge about healthy and unhealthy diet. Our conclusions at the end of this study is that the family with children in primary schools do not get involved in oral/dental health education. PMID- 12638295 TI - [Professional guidance problems in spastic hemiparesis school children]. AB - People with physical handicap adapt themselves with more difficulties to professions that require manual abilities. For these youngsters, the professional guidance plays a major role. Their desire is to become independent from the social point of view, independent which is linked with a professional activity. If the youngsters with physical handicaps are correctly orientated, they become socially independent and they perform their professional duties with great correctness. In order to fulfill this goal we must understand the nature of the motric problems. This knowledge is obtained by using a series of tests that allows us to evaluate the degree of motric handicap. The physical difficiency, followed by the integration in a special school, may result in personality problems. Briefly, the motric handicap problem must be discussed in a complex, following medical aspects but also professional, psychic and social aspects. PMID- 12638296 TI - [Determination of several erythrocyte enzymes activity in patients with different tumors of the oral cavity]. AB - Considering the influence on the molecular level of the neoplasic factors, upon several proteins, nucleic acids, one can say that some of the oncogenesis determinants are represented by genetic mutations. Free radicals, including also some organic peroxides are considered as tumour promoters, although the exact mechanism of this process in still unknown. The neoplasic disease is characterized generally by disorders of the control processes, including the one displayed on the subcellular level. Considering the enzymatic changes occurred in erythrocytes and determined by the disturbances of membrane permeability, we evaluated the response of several aggressions at the erythrocyte level, in case of maxillo-facial tumours. Our results show increase of the LDH, G-6-P-DH activity and decrease of catalase activity within the erythrocyte. PMID- 12638297 TI - [Gelatinases in peri-implantation stage]. AB - The periimplantation stage in mice is marked around the 5th day after mating by blastocyst presence in the emdometrium. The events that are marking the implantation event are focused on the evolution of the extracellular matrix around the mezenchime cells that undergoes decidualization. The inductive process is guided by gelatinases and the purpose of our study is to demonstrate their presence at this level. PMID- 12638298 TI - [Preventing methods of prosthetic stomathopaties concerning patients with partial removable prosthesis]. AB - Partial removable prosthesis is the most frequently therapeutical solution applied to patients aged more than forty years and which were diagnosed with extended partial edentations. The purpose of our study is to estimate oral tissues diseases installment to subjects having partial removable prosthesis, this patients being exposed on contacting stomatopaties. We developed this study on the clinical examination of a group of forty subjects partial removable prostheded, three or four year ago, respectively 23 woman and 17 men aged between 40 and 75 years. The oral hygiene, caries incidence, clasps fractures were evaluated, it was registered the aspect of the acrylic resin of the base, and also the inflammation degrees of the oral mucousa. A high level of oral hygiene statement diminuates the installment of the whole series of post-prothetical involvements, also decreasing the stomatopathies appearance frequently. PMID- 12638299 TI - [Xanthine derivative compounds potential activity in inflammatory process]. AB - We studied the potentially anti-inflammatory action of some new xanthine compounds, theophylline derivatives with 7-[2-hydroxy-3-(4-acetamido)-phenoxy propyl]-8-R-1,3-dimethyl-xanthine and 8-R-1,3-bis-(1,3-dimethyl-xanthin-7yl)-2 hydroxy-propane structure substituted in 7 and 8 position. Almost all studied compounds reduced the inflammatory edema, the effect being comparable or highest theophylline. The antiinflammatory effect of theophylline intensified with the introduction in 7 position the theophylinyl radical and in 8 position the bromo, nitro, pirolidinyl, piperidinyl, morfolinyl, imidazolyl, 3,5-dimethyl-pirazolyl, 3-methyl-5-oxo-pirazolyl radicals. PMID- 12638300 TI - [Multiple perfusion defects detected by scintigraphy]. AB - Pulmonary embolism is a cause of death and the most common pathological condition involving the lungs of hospitalized patients. Early detection can rise prognosis of survival. In this case presented below, a patient suspected of pulmonary embolism was sent to our department, for assessing the lung perfusion, performing the pulmonary perfusion scintigram with 99mTc-MAA for establishing a diagnosis. Lung imaging is a noninvasive technique without patient discomfort that objectively can depict the distribution of the radiopharmaceutical throughout the lungs. We conclude that multiple non segmental defects with no radiological abnormalities state that the probability of pulmonary embolism is lower than 20%. PMID- 12638301 TI - [Consideration on serotoninergic syndrome based on a clinical situation ]. AB - The diagnosis criteria for serotoninergic syndrome was first time proposed in 1991 by Sternbach, mixing clinical, biological and historical features. Drugs inducing serotonin accumulation by decreasing the synaptic recapture or increasing the serotoninergic transmission are responsible for the onset of this syndrome even in small amounts. We are describing a case of a young patient developing a comatose stage of serotoninergic syndrome as a result of Amitriptylinum and Sertralinum poisoning. PMID- 12638302 TI - [Adult celiac disease(gluten enteropathy]. AB - Celiac sprue is a disease in which there is malabsorption of nutrients by that portion of the small intestine which is damaged, a characteristic though not specific lesion of the small intestinal mucosa and prompt clinical improvement following withdrawal of certain cereal grains from the diet. We present a 41 years old patient with a history of diarrhea, treated as enterocolitis for several years, in which the gluten-free diet brought a clinical improvement. PMID- 12638303 TI - [Osteomyelitis, a cause of prolonged fever in adults]. AB - A 35 years old male patient with an unexplained prolonged fever, after serial unsuccessful investigations in 3 different services, suffers a fracture of the lower third of the right femur produced by a minor trauma. This raises the suspicion of a pathological fracture. The surgical intervention and the microbiological exam confirms the existence of an osseous infection with Gram negative germs, and the clinical course is uneventful after stabilization with an external fixator and appropriate antibiotic therapy. Osteomyelitis, although rare in adults, should be included in the differential diagnosis of any prolonged fever of unknown origin in adults. PMID- 12638304 TI - [Methods of advanced purification-the challenge for biosynthetic antibiotics industry]. AB - Reactive extraction, permeation through liquid membranes and direct extraction are some of the new techniques applied for separation and advanced purification of biosynthetic antibiotics. Compared with the conventional separation techniques, the main advantages of these extraction methods are: high separation efficiency, the avoidance of antibiotics chemical and thermal inactivation, high purity of obtained antibiotics. Furthermore, using reactive extraction or permeation through liquid membrane, the antibiotics can be selective separated from their biosynthesis precursors or from the secondary biosynthetic compounds. This paper is a review on separation of Penicillins and Erythromycin by means of these extraction techniques, being underlined their advantages, applications and problems concerning the separation process scale-up. PMID- 12638306 TI - [An invasion of insect Simulium columbaczese (Diptera, Simuliidae) described in an old Romanian chronicle]. AB - The oldest description of an invasion of Simulidae on the present territory of Romania, containing unique data for the history of parasitology in the Danube valley, has been discovered by the author in an anonymous chronicle "Istoria Tarii Romanesti de la Octombrie 1688 pana la Martie 1717 (History of Wallachia from October 1688 to March 1717), written at the Court of the Prince of Wallachia, Constantin Brancoveanu. In the last decade of the XVIIth ctry, the Prince, at the head of his army, had to march in order to defend the Wallachian Danube border in the area of its gorges. The chronicle describes the delay caused on the way by an invasion of "poisonous" flies, attacking both animals and people. This description is detailed enough to establish that it is regarding an invasion of Simulium columbaczense, insect present in the area even nowadays. The development of these insects is favourised by numerous rivulets coming down mountain slopes of the Danube. PMID- 12638305 TI - [Bifocal non-union of the forearm as of consequence of an open fracture type 3 B a case report]. AB - We present a 26-year old patient who suffered an accident while at work, his forearm being almost completely separated because of an electrical circular saw. Admitted at our hospital 8 hours after the accident occurred the forearm was temporarily stabilized by means of k-wires and the radial artery and vein as well as the median nerve were anastomosed. PMID- 12638307 TI - [Causes of late renal transplant dysfunction]. AB - Renal transplantation is now established as the therapy of choice for end-stage renal failure. The causes of renal allograft loss have changed with the introduction of new immunosuppressive agents. In the pioneer era of transplantation most renal allografts were lost during the first year after transplantation due to acute rejection episodes. Nowadays, chronic allograft nephropathy became the leading cause of graft loss. The causes of chronic allograft nephropathy can be divided into alloantigen-dependent and alloantigen independent factors. Acute rejection episodes and histoincompatibility have a significant influence on late graft function. Although alloantigen-related injury is of major importance, alloantigen-independent factors also play a significant role in the progression of chronic allograft nephropathy. Prolonged ischemia time leads to induction of inflammation, resulting in fibrotic scarring or proliferation of mesenchymal cells. Donor related factors such as donor brain death, age, nephron number and gender have a definite impact on late allograft function. Posttransplant complications such as hypertension, metabolic factors and viral infections contribute to accelerated deterioration of the functional units of the kidney. Immunosuppressive agents such as cyclosporine and tacrolimus may cause vasoconstriction and decreased glomerular filtration rate. Moreover, non-compliance is one of the most important risk factor for chronic allograft loss. Regardless of the cause of the initial injury the pathophysiology of chronic allograft nephropathy seems to be the same. All these factors can induce endothelial injury, which leads to increased expression of cytokines and growth factors resulting in proliferative processes, remodelling and scarring of the graft. This paper reviews current knowledge about risk factors and their effect on long-term renal allograft function. PMID- 12638308 TI - [Hypertension and menopause]. AB - Cardiovascular disease is the leading cause of death among men and women. The cardiovascular risk of the women reaches the men of same age after the menopause. Hypertension is one of the most powerful and prevalent contributor of ahterosclerotic cardiovascular disease. Hypertension affects more men than women until 55 years of age, but after 55 the percentage of women is higher. Estrogen deficiency has been linked to the rapid progression of cardiovascular risk of women after menopause. The biological background of protective effects of estrogen is not fully, but mainly understood yet. It may involve direct effects on blood vessels through modulation of endogen vasoconstrictors and vasodilators, accompanied by systematic effects of the hormone. The main effect of these influences protects the cardiovascular system. After menopause, loss of this protection may unmask a population of women prone to hypertension, who would be at higher risk for cardiovascular disease. The guidelines make no differences apart form gestational hypertension between men and women. This review would like to focus on the clinical relevance and characteristics of postmenopausal hypertension according to the literature. PMID- 12638309 TI - [Sphincter of Oddi dysfunction--prolonged medical therapy or early endoscopic sphincter ablation]. AB - INTRODUCTION: Sphincter of Oddi dysfunction is a real challenge from both diagnostic and therapeutic point of view. PATIENTS AND METHODS: In the last two years the authors have performed ERCP and EST in 29 patients with positive evocative test results, who had important enzyme elevations and/or did not respond to prolonged medical treatment. RESULTS: Endoscopic findings were positive in 25/29 patients (86.2%): 8 adenoma of p. Vateri, 17 papillitis were identified, and in 4 cases the papilla was intact. Histopathology obtained in 12 patients supported the diagnosis. In 6 patients, who underwent a postpapillotomy evocative test, after an average of 10 months follow up the results have been converted from positive to negative response in all but two cases. The two patients continued to have abdominal symptoms with persistent positive provocation tests because of restenosis, were treated with repapillotomy. CONCLUSIONS: The Debray and Nardi tests are useful screening tests for hypertonic biliary or pancreatic dyskinesia. Structural endoscopic and histological findings are frequent already in the functional cases. Early sphincter ablation should be considered in failure of medical therapy for preventing the transformation of this functional disorder into an organic, potentially precancerous state. PMID- 12638310 TI - [Comparative study of combined local treatment (sulfadimidine, metronidazole and nystatin) and the standard monotherapy in uncomplicated bacterial vaginosis]. AB - Comparative, in vivo, human, prospective, single blind, clinical and microbiological diagnoses based and randomised study of the treatment of uncomplicated bacterial vaginosis with two forms of combined (metronidazole + nystatin + sulfadimidin) vaginal suppositories (laminated and mixed containing the same ingredients) and the standard preparations available in the Hungarian market (Dalacin vaginal cream and Klion vaginal suppository). The examinations involved 60 volunteers and were performed in the Gynecological Outpatient Clinic of the Council of Erd, the microbiological samples were examined at Saint Rokus Hospital in Budapest. The combined treatment was better tolerated and resulted in normal vaginal pH significantly more often at the same rate of recovery. The combined treatment is simultaneously effective in cases of the most prevalent coinfections too. PMID- 12638311 TI - [Isolated ovarian abscess in the puerperium]. AB - Isolated ovarian abscess is the infection of the ovary without tubal involvement. A number of predisposing factors have been identified. The appearance after spontaneous vaginal birth is rare. The clinical presentation is varied, the symptoms are not specific. Proper diagnosis necessitates the complete evaluation of clinical signs, laboratory tests and imaging techniques. The effective treatment is mostly operative. The authors present the detailed management of a postpartum patient with an isolated abscess of the right ovary. Antenatal identification and treatment of cervico-vaginal infection along with strict antiseptic way of care during labor and delivery may result in less prevalent and less severe postnatal maternal inflammatory complications. PMID- 12638312 TI - [The LDL receptor family]. AB - The members of the LDL receptor family are structurally related endocytic receptors. Our view on these receptors has considerably changed in recent years. Not only have new members of the family been identified, but also several interesting observations have been published concerning the biological function of these molecules. The LDL receptor family members are able to bind and internalize a plethora of ligands; as a consequence, they play important roles in diverse physiological processes. These receptors are key players in the lipoprotein metabolism, vitamin homeostasis, Ca2+ homeostasis, cell migration, and embryonic development. Until recently, LDL receptor family members were thought to be classic endocytic receptors that provide cells with metabolites on one hand, while regulating the concentration of their ligands in the extracellular fluids on the other hand. However, recent findings indicate that in addition to their cargo transport function, LDL receptor family members can act as signal transducers, playing important roles in the development of the central nervous system or the skeleton. Better understanding of physiological and pathophysiological functions of these molecules may open new avenues for the treatment or prevention of many disorders. PMID- 12638313 TI - [Incidence of thyroid disorders in children in Hungary]. AB - Author gives information to the home tendencies of incidence of some important thyroid disorders in childhood mainly on the basis of his studies done with his coworkers. Results are compared with the standpoints raised since Chernobyl accident. It is verified on the basis of cca one million neonatal TSH screening that the frequency of congenital hypothyroidism in Hungary corresponds to those of European countries in the last 17 years. Changes connected with Chernobyl has not occurred. It deals with incidence of Graves' disease as the main form of hyperthyroidism relying upon TSH receptor antibody positivity. The yearly distribution of the 176 proved cases shows a slow increasing trend in the same period of time (17 yrs). In this radiation does not play a role. The paper treats of the topic of iodine deficiency with of high priority. Author draws the attention again to the fact that the iodine nutrition in significant part of the population is behind of optimum in harmony with the results of some other national studies also according to their own three thousand urinary iodine tests and more than four thousand thyroid sonographies. This iodine deficiency should be regarded as a potential risk from the viewpoint of radiation hazard! While enlargement of thyroid was often found by sonography, the formation of nodules only rarely in contrast to those of the Chernobyl area. The frequency of thyroiditis, especially in puberty increases too, but the degree of it falls far behind from those of published around of Chernobyl. The incidence of differentiated thyroid carcinoma--according to the available data--moderately increases as well, without any connection between the residence and the geographical distribution of radiation pollution. The epidemiological, clinical, and histological characteristics of the Hungarian children are similar to the so called sporadic (non-radiation induced) form and different from the cases of the Chernobyl area. PMID- 12638314 TI - [Endobronchial ultrasonography]. AB - Endobronchial ultrasonography is an imaging method which has been developed during the last decade. It can be performed by applying tools that are used today during conventional bronchofiberscopy. This method is recommended for identifying processes that affect the bronchial wall and the mediastinal lesions next to central airways as well as for localizing peripheral lung lesions. The authors present a compilation of publications and summarize their own experiences based on 44 cases of application of this method. PMID- 12638315 TI - [First case of successful pregnancy after preimplantation genetic diagnosis]. AB - Preimplantation genetic diagnosis is introduced for prevention of genetic disorders. The combination of in vitro fertilization technique and single cell molecular genetic diagnosis allows only unaffected embryos to be selected for embryotransfer, providing a healthy pregnancy and so also avoiding the need for its possible termination. CASE REPORT: The authors report the first successful case of the clinical application of preimplantation genetic diagnosis in Hungary, resulting the birth of an unaffected baby. Preembryonal biopsy and subsequent fluorescent-PCR analysis of the blastomeres taken from the preembryos of a woman who is a carrier for Duchenne muscular dystrophy was performed. Sexing of the preembryos by X and Y chromosome specific primers were performed and three female preembryos were found. RESULT: All three preembryos were transferred into the uterus, which resulted a singleton pregnancy, resulting the birth of a healthy female baby. The authors are offering preimplantation genetic diagnosis with sexing for couples, who are at risk of having a child with X-linked diseases, or are carriers for the delta-F508 mutation of cystic fibrosis. CONCLUSIONS: The application of this diagnostic tool is also planned for other monogenic disorders. PMID- 12638317 TI - [Commencement address of the 234th academic year]. PMID- 12638318 TI - [Recollections about the role of "Extrapulmonary TBC meetings" in the prevention of extrapulmonary tuberculosis]. PMID- 12638316 TI - [Ursodeoxycholic acid treatment in intrahepatic cholestasis of pregnancy. A case report]. AB - INTRODUCTION: Intrahepatic cholestasis of pregnancy is a rare disorder of unknown origin with a symptomatically distressing maternal course with pruritus and icterus as the chief complaints. The disease poses little medical risk to the mother, but poses significantly risk to the fetus of perinatal mortality, preterm delivery, fetal distress, and meconium staining. Beside the pruritus and the high level of bilirubin in the serum the increased values of parameters showing the liver function and of alkaline phosphatase activity are characteristic. Ursodeoxycholic acid is the only therapeutic modality whose effectiveness was proven by controlled trial. CASE REPORT: The authors demonstrate the case of a 29 year old woman who developed intrahepatic cholestasis with the characteristic clinical and laboratory findings at the 33rd week of her pregnancy. The clinical complains disappeared and the liver function tests were better after the applied ursodeoxycholic acid treatment. The delivery was carried out without complications with a healthy newborn baby. DISCUSSION AND CONCLUSION: On the basis of the data in the literature and of this case report the authors recommend the ursodeoxycholic acid treatment in the management of intrahepatic cholestasis of pregnancy. PMID- 12638319 TI - Perspectives in cardiac ultrasound. PMID- 12638320 TI - Clinical and echocardiographic features in patients with dilated cardiomyopathy: wave intensity and diastolic abnormality analysis. AB - OBJECTIVE: The pathophysiology of dilated cardiomyopathy is not fully understood, and among the problems which need to be solved is the lack of an adequate classification which could help to predict the survival of patients in the end stage of the disease. The criteria used today, based on clinical symptoms and invasive and noninvasive assessment of the heart function, are far from satisfactory. Estimation of maximal oxygen consumption is a time-consuming procedure and more descriptive of the global cardiopulmonary situation rather than being able to provide specific answers. Loading conditions seem to be the key to the problem. The state of vascular resistance and systolic performance of the heart can be defined using the method known as "wave intensity" (WI), which represents the relation of the pressure estimated noninvasively to the flow inside the vessel. This study presents first experiences with wave intensity in the Deutsches Herzzentrum Berlin in patients suffering from end-stage dilated cardiomyopathy (DCM), a group of ambulatory patients after heart transplantation (HTx), and in healthy volunteers. The study was performed in supine and upright position. PATIENTS: Out of 182 patients waiting for transplantation, thirty-one (17%) ambulatory patients suffering from dilated cardiomyopathy comprised the first group (Group A). Their mean age was 53 +/- 14.2 years and 16 were men. Maximum MV O2 was 16.1 (+/- 4.57) mg/kg. Mean PAP and capillary pressure were 26.1 (+/- 11.91) and 16.8 (+/- 9.81) mmHg, respectively. Cardiac index was 2.3 (+/- 0.58) l/min/m2. The second group (Group B) consisted of 56 ambulatory patients after heart transplantation (mean 3.8 +/- 3.35 years) who did not suffer from rejection. Their mean age was 49.9 +/- 15.18 years and 47 were men. Normal sinus rhythm was an inclusion criterion for the study. The control group consisted of 15 healthy volunteers with a mean age of 35 +/- 12 years and 2 were men. METHOD: Real time wave intensity (WI), Doppler mitral filling and noninvasive cardiac output were recorded and analyzed using the ALOKA 5.500 machine equipped with a wave intensity real time analysis system. Wave intensity was defined noninvasively as the product of delta U and delta V where delta U was vascular diameter change (representing pressure change) and delta V was the velocity inside the vessel. The measurements were done in real time during constant short time intervals. The WI data were obtained from the right common carotid artery in all patients. RESULTS: The 1st and 2nd WI peaks were significantly lower in supine position in the DCM group and rose significantly in upright position when compared to the data from patients after transplantation and volunteers. The values of 1st and 2nd peak in supine position in DCM were respectively 4606 (+/- 2283) and 1483 (+/- 675) and upright 1st and 2nd were 5405 (+/- 3432) and 202 (+/- 1726). The values of 1st and 2nd peak in the Tx group were significantly higher at rest: 10,743 (+/- 6290) and 3195 (+/- 2127), respectively, and were not significantly lower after upright maneuver. The highest values of both products (1st and 2nd) of WI were observed in healthy volunteers 14,632 (+/- 6293) and 14,153 (+/- 6969) and were significantly higher when compared to the DCM group (p = 0.01). Cardiac output in the supine position was significantly lower in the DCM patients and fell after position was changed to erect. In the Tx group, cardiac output was normal at rest and didn't fall significantly in the upright position. The DCM and transplant patients showed abnormalities in diastolic heart function after changing to the erect position (restrictive pattern of mitral filling); however, more severe filling abnormalities were found in the DCM patients. CONCLUSION: Our results demonstrate that patients suffering from DCM are less tolerant of arthostatic stress than patients after heart transplantation. The slight increase in wave intensity in the DCM group hints at stronger activation of autonomic vascular control, leading to a stiffening in the arterial system, which possibly protects the central nervous system from hypoperfusion. The diastolic alteration was probably the response of diseased myocardium to increased requirements of the peripheral system after changing the position of the patients. This diastolic alteration, although less pronounced, was noticed in the transplanted group, but was not seen in healthy individuals. Therefore, we conclude that wave intensity is a new, powerful marker providing information not only about left ventricular systolic function but also about mechanoelastic properties of the arterial system integrated with elastic properties of the myocardium. PMID- 12638321 TI - [Evaluation of the respiratory system in patients with heart failure based on spiro-ergometric exercise test parameters]. AB - The objective of this study was to assess the ventilatory parameters of cardiopulmonary exercise test (CPX) in patients with moderate and severe heart failure (HF). MATERIAL AND METHODS: We investigated 110 patients (pts) (mean age 52.1 +/- 11.1 years) with chronic HF due to dilated cardiomyopathy (42 pts) or coronary artery disease (68 pts). Resting spiro-metry, CPX on a treadmill (modified Naughton protocol, 83 pts, or modified Bruce protocol, 26 pts) and echocardiography were performed. The following parameters were analyzed: left ventricle ejection fraction (EF), forced expiratory volume in 1 second (FEV1) and forced vital capacity (FVC). Both FEV1 and FVC are expressed as a percentage of predicted normal values (FEV1-%N, FVC-%N), FEV1/FVC ratio, rest and peak parameters of CPX ratio of physiological dead space to tidal volume (VD/VTrest, VD/VTpeak), respiratory rate (RRrest, RRpeak), minute ventilation (VErest, VEpeak), tidal volume (VTrest, VTpeak), endtidal Pco2 (PETCO2rest, PETCO2peak), endtidal Po2 (PETO2rest, PETO2peak), ventilatory equivalent for carbon dioxide (VE/VCO2rest, VE/VCO2peak) and oxygen (VE/VO2rest' VE/VO2peak), exercise time (T), peak oxygen uptake VO2peak' VO2peak%N, heart rate reserve HRR, peak heart rate HRpeak%N. After the mean follow-up period of 28.5 +/- 12 months, the patients were divided into groups: Group I: n = 78 pts who survived without heart transplantation (HTX) and Group II: n = 32 pts; n = 20 pts who died while on the transplant list; n = 12 pts who underwent urgent HTX. RESULTS: The patients in the group II had lower EF, more reduced peak oxygen consumption and shorter duration of exercise. We found that FEV1, FVC, FEV1-%N, FVC-%N were more severely impaired in group II. The patients in group II also had lower PETCO2rest and higher PETO2rest. There were no differences in the FEV1/FVC ratio, VD/VTrest, RRrest, VTrest, VErest, VE/VCO2rest, VE/VO2rest between the groups. The value of VD/VTpeak, VE/VCO2peak, VE/VO2peak, PETCO2peak were significantly higher in group II when compared with group I, but the values of PETO2 were lower. Despite a shorter duration of CPX in group II there were no differences in HRpeak%N, HRR, RRpeak, VTpeak, or VEpeak, between the groups. CONCLUSION: At rest the patients with more severe HF appear to have a more restrictive pulmonary pattern and impaired pulmonary gas exchange. During exercise, the patients with severe HF have an increased dead space in the lung; more impaired pulmonary gas exchange and increased ventilatory response at a given workload. PMID- 12638322 TI - [Prognostic value of results from clinical tests, echocardiographic, electrocardiographic and spiro-ergometric exercise test examinations in patients with heart failure]. AB - The aim of this study was to assess the importance of the non invasive procedures in determinating prognosis of patients with heart failure (HF). MATERIAL AND METHODS: We investigated 110 patients with a mean age of 52.1 (65 patients--59%) with chronic (HF) due to coronary artery diseases and dilated cardiomyopathy (45 patients--41%). All the patients underwent baseline evaluations including a clinical status, echocardiography, Holter monitoring and a cardiopulmonary exercise test (CPX). After the mean follow-up period of 28.5 +/- 12 months, the patients were divided into 2 groups: Group I--in which 78 patients survived without heart transplantation (HTX) and Group II--in which 20 patients died and 12 underwent HTX. The relationship between the variables derived from noninvasive procedures was assessed using multivariate Cox proportional hazards models. RESULTS: Compared to group I, Group II had a significantly higher number of patients that were classified into NYHA class III. In addition, a third heart tone was identified more often in group II and an increased number of HF etiology was due to ischemia. Electrocardiographic recording showed an increased frequency of left bundle branch block and complex ventricular arrythmias. Markedly increased values of WSLK, OKSLK, WLP, OKRLK and FW were found in group II, as compared to group I. Cardiopulmonary exercise tests in group II revealed lower values of T, VO2AT, SBPpeak, VO2peak, VO2peak%N, PETCO2peak as well as VD/VTpeak, VE/VO2peak, and VE/VCO2peak. After multivariate analysis lope VE/VCO2peak (p = 0.009), peak oxygen uptake VO2peak-%N (p = 0.035), (p = 0.036), left ventricle end systolic volume (p = 0.045) and left bundle branch block (p = 0.046) were independent predictors of survival without heart transplantation. CONCLUSION: On multivariate analysis by the Cox proportional hazards method, the variables: VE/VCO2peak, VO2peak%N, etiology--CAD, OKSLK, and left bundle branch block were independent prognostic predictors. The echocardio-graphic evaluation play a significant role in the assessment of prognosis in patients, with HF. The ejection fraction of the left ventricle was not confirmed as an independent prognostic factor in HF. PMID- 12638323 TI - [Effect of low dobutamine doses dobutamine on left ventricular diastolic flow parameters in subjects with a low ejection fraction]. AB - A low dobutamine dose (DOB) improves the systolic function of impaired myocardium. There are some data suggesting the improvement of left ventricular (LV) diastolic function as well. The aim of this work was to assess the influence of DOB on some left ventricular diastolic flow parameters: inflow curve profile and early inflow wave propagation velocity (Prop). MATERIAL AND METHODS: 33 persons (7 women and 26 men) aged 35-74, with a mean age of 53 and with an ejection fraction (EF) < 35% were examined. DOB was administered in doses of 5 and 10 micrograms/kg/min every 3 minutes. At the beginning of the examination (D 0) and at the end (D-1), the LV contractility and the following inflow parameters were assessed: E and A waves velocities, E/A-ratio on mitral orifice level (U) and at the half of LV (1/2 LV) and Prop. RESULTS: DOB increased the heart rate from 69.4/min to 74.4/min (p < 0.01) and improved the LV contractility in every case. We observed no significant changes of the E wave velocity influenced by DOB on U (D-O: 0.7 +/- 0.22 m/s; D-1: 0.68 +/- 0.2 m/s) and in 1/2 LV, or no E/A changes (D-O: 1.24 +/- 0.71; D-1 1.21 +/- 1.08 on U and D-O: 1.68 +/- 1.06; D-1: 1.64 +/- 1.16 in 1/2 LV). Statistically significant differences were recorded only between A in D-O and A in D-1 on both levels (p < 0.05): D-O was 0.66 +/- 0.21 m/s and D-1 was 0.71 +/- 0.22 m/s on U and D-O was 0.422 +/- 0.2 m/s and D-1 was 0.47 +/- 0.2 m/s in 1/2 LV. Significant Prop shortening was ascertained: from 39.4 +/- 10.9 cm/s to 45.4 +/- 16.1 cm/s; p < 0.01. The correlation registered at rest between Prop and EF (R = 0.4) and Prop and contractility index (R = -0.52), was recorded at a similar level after DOB as well. No correlation between Prop and heart rate was seen. CONCLUSIONS: 1. Dobutamine in low doses in subjects with impaired LV causes no significant heart rate acceleration and no progression in contractility disorders, improves LV diastolic function, expressed by early diastolic blood flow towards the apex (inflow propagation) acceleration. 2. Dobutamine in low dose increases mitral A wave velocity caused by improvement of LV diastolic function and/or improvement of LA contractility. PMID- 12638324 TI - [Reproducibility of spiro-ergometric exercise test parameters in patients with heart failure]. AB - The aim of the study was to determine the degree of reproducibility of parameters of cardiopulmonary exercise (CPX) tests in patients with chronic heart failure (HF). MATERIAL AND METHODS: We investigated 30 patients (7 women, 23 men) with ages ranging from 44 to 68 and a mean age of 56 +/- 14.5 years with HF due to dilated cardiomyopathy (5 patients) and coronary artery diseases (25 patients). The mean ejection fraction was 24 +/- 6.2%. All the patients were symptomatically stable during the preceding 4 weeks. After a preliminary test, the patients underwent two treadmill CPX tests using modified Naughton protocol with a 1-week interval between tests. CPX tests were performed at the same time of the day, using the same equipment and the patients were receiving constant medication. The reproducibility was assessed for: Rest heart rate (HRrest), oxygen uptake at the anaerobic threshold (VO2AT) and peak exercise (VO2peak), carbon dioxide production (VCO2peak), ventilatory equivalent of carbon dioxide (VE/VCO2peak) and oxygen (VE/VO2peak), end-tidal partial pressure for carbon dioxide (PETCO2peak), exercise time (T) and respiratory exchange ratio (RER). RESULTS: The reproducibility was found for all analyzed parameters of CPX tests: HRpeak (p < 0.0001), VO2AT (p < 0.0001), T (p < 0.001), HRpeak (p < 0.0001), VO2peak (p < 0.0001), VO2peak%N (p < 0.0001), VE/VCO2peak (p < 0.0001), VE/VO2peak (p < 0.0001), PETCO2peak (p < 0.0001), RER (p < 0.004). CONCLUSION: On adherence to standardized conditions, an excellent reproducibility exists for the most important parameters of cardiopulmonary exercise tests. CPX tests provide a useful and objective method of assessing functional capacity and diseases progression in patients with heart failure. PMID- 12638325 TI - [Prognostic value of a model using restricted left ventricular filling for prediction of developing heart failure after myocardial infarction]. AB - The prognostic significance of restrictive pattern (RP) of mitral flow in myocardial infarction (MI) has yet not been defined. The aim of the study was to assess the relationship of a RP in MI patients at discharge with the development of left ventricular (LV) dilatation and clinical symptoms of heart failure (HF) after one year. METHODS: 2D and Doppler Echocardiography was performed on 88 patients (mean age 53.8 +/- 11) at discharge after their first MI and then one year after that MI. Patients were classified according to Doppler examination at discharge into 2 groups: Group I-RP (n = 9) with peak E to A ratio E/A > or = 2 or E/A > or = 1.5 and deceleration time DT < or = 140 ms, Group II--non restrictive pattern nRP (n = 79). In predicting the development of HF the following variables were analysed using Cox proportional hazard model: MI location and extension (Inf, Ant, non-Q, Q-wave), reperfusion defined by non invasive features (R/+/, R/-/), Killip-Kimball class (K-K), end-diastolic volume index (EDVI), end-systolic volume index (ESVI), ejection fraction (EF), wall motion score index (WMSI) and restrictive pattern of mitral flow presence (RP). RESULTS: HF developed in 23 patients: 8 pts were in RP-group (89%), while 15 pts were in nRP-group (19%) (rr = 4.68 p < or = 0.001). In the univariate Cox analysis, the following variables were associated with higher incidence of HF: Q MI (rr = 5.66 p < or = 0.05), R- (rr = 4.96 p < or = 0.001), K-K > or = II (rr = 3.13 p < or = 0.01), EDVI > or = 80 ml/m2 (rr = 5.85 p < or = 0.001), ESVI > or = 40 ml/m2 (rr = 4.07 p < or = 0.001), WMSI > or = 1.4 (rr = 2.79 p < or = 0.01) and RP (rr = 4.6 p < or = 0.001). However, the multivariate Cox analysis revealed only three significant independent predictors of HF: EDVI > or = 80 ml/m2 (rr = 6.27 p < or = 0.001), R- (rr = 4.74 p < or = 0.01) and RP (rr = 3.2 p < or = 0.05). The cumulative predictive power of RP in connection with the selected parameters (RP + K-K > or = II, RP + EF < or = 45%, RP + WMSI > or = 1.4) was higher for RP + K-K > or = II (rr = 4.61 p < or = 0.001) and especially for RP + WMSI > or = 1.4 (rr = 5.06 p < or = 0.001). CONCLUSION: The restrictive pattern of LV filling in MI pts at discharge is an independent predictor of heart failure development. The assessment of both diastolic and systolic LV functions parameters increases prediction power. PMID- 12638326 TI - [Level of angiotensin II and aldosterone in plasma is often elevated in patients with heart failure treated with converting enzyme inhibitors--preliminary results]. AB - In large randomised trials, ACE inhibitors (ACEI) have been shown to reduce mortality, morbidity and improve quality of life in patients (pts) with congestive heart failure. However, long-term prognosis of patients in the community remains poor. It has been suggested that one of the reasons may be inadequate neuroendocrine suppression with current treatment strategies. To address this issue we measured plasma levels of angiotensin II (AII) and aldosterone (Ald) in 41 patients (36 males, mean age 52 +/- 2 y) referred to our department for diagnostic evaluation, who were treated with clinically appropriate doses of ACEI. The mean angiographic left ventricular ejection fraction was 22 +/- 8%, left ventricular end diastolic diameter was 72 +/- 10 mm, and NYHA class was 2.6 +/- 0.7. Plasma levels of All and Ald were measured by radioimmunoassay. They did not differ significantly in comparison with the control group of 5 healthy individuals (4.8 +/- 8.2 pg/ml vs 4.1 +/- 3.2 pg/ml for angiotensin II and 129 +/- 93 pg/ml vs 78 +/- 29 pg/ml for aldosterone). A high variability of the results was seen between the individual patients. Full suppression of All (< 2.0 pg/ml) was achieved in 21 patients (58%), 10 pts (28%) showed intermediate levels (2.0-10.0 pg/ml), and 5 patients (14%) demonstrated markedly increased All levels (> 10 pg/ml). Full suppression of Ald (< 80 pg/ml) was seen in only 15 patients (37%), 14 patients (34%) had intermediate levels (80 140 pg/ml) and 12 patients (29%) showed high plasma levels of Ald (> 140 pg/ml). There was a weak, but significant, correlation between All and Ald levels in the study group (r = 0.49, p < 0.05). These preliminary results suggest inadequate neuroendocrine suppression in a substantial proportion of patients, despite using clinically relevant doses of ACE inhibitors. The determination of All and Ald levels may be a helpful tool in monitoring the efficacy of treatment in CHF and may help identify patients who would benefit from other treatment strategies. PMID- 12638327 TI - [Echocardiographic evaluation of left atrial function in dilated cardiomyopathy with restricted and non-restricted Doppler transmitral flow]. AB - Left atrial (LA) function is of great importance in left ventricular (LV) filling. There is evidence that echocardiographic Doppler evaluation of transmitral flow, routinely used for LV filling estimation, is dependent on LA function. Information regarding the relation of LA size and function to transmitral flow in heart failure is limited. We used 2D echocardiographic acoustic quantification methods to assess LA function in patients with dilated, (non-ischemic) cardiomyopathy (DCM) and a control group. The DCM group was divided into 2 subgroups: Group 1-with restrictive LV Doppler filling pattern-18 patients (DCM-R) and Group 2-with non-restrictive LV Doppler filling pattern-11 patients (DCM-NR) with similar heart rate, age and degree of mitral regurgitation. LA maximal area, total emptying fraction and absolute and fractional area change during rapid emptying and atrial contraction were calculated. The LA was enlarged only in DCM-R. Both DCM groups had decreased total emptying fractions and rapid emptying area changes compared to controls. An enlarged LA area and more decreased total emptying were found in DCM-R with high LV filling pressures compared to DCM-NR. The restrictive group had a significantly smaller LA rapid emptying area change, as well as a smaller LA area change and emptying fraction during atrial contraction compared to DCM-NR. Within < or = 2 hrs after the echocardiography study, cardiac catheterization was performed in the DCM group. We found significantly higher LV filling pressures and lower LV ejection fractions in DCM-R compared to DCM-NR. Significant correlations were found between LA function and invasive parameters like capillary wedge and LV enddiastolic pressures and LV EF. CONCLUSION: Patients with DCM-R had significantly enlarged LA areas with more depressed total emptying fractions and smaller LA area changes during contraction compared to DCM-NR. Thus, left atrial function plays an important role in LV filling and its dysfunction can be a marker of poor prognosis. PMID- 12638328 TI - Magnetic resonance imaging of the heart. PMID- 12638329 TI - [Use of magnetic resonance spectroscopy in cardiology. State of the art]. AB - Magnetic resonance imaging of tissues and organs, specifically the use of magnetic resonance specroscopy (MRS) for examination of metabolism in vivo, is a relatively new modality with a very dynamic development and a promising future. In the past few years, magnetic resonance imaging (MRI) has been used more widely in cardiology. A certain stereotype has been broken that this modality is dedicated strictly for neuroradiology. The technological advances in MRI include introduction of new systems with short acquisition times and software for imaging of the heart and large vessels (cardiac package). Potent and fast gradients in new MR systems make it possible to use cardiac MRS in clinical practice. The introduction of in vitro spectroscopy is an important step in the identification of the metabolic components of the myocardium, just as the in vivo spectroscopy was important in neuroradiology. This method has been used over many years for basic science, and only now is it being used widely in clinical practice. In the examination of heart metabolism two types of MRS are frequently used: phosphate (31PMRS) and proton (1HMRS). The phosphate spectroscopy examines the composition and metabolism of high energy compounds, intracellular pH and indirectly provides information about glucose metabolism. Proton spectro-scopy determines lipid (Lip), lactate (Lac) and high-energy compounds--creatine (Cr) levels. Heart MRS examinations are performed in many clinical centers around the world. In this review about MRS, the authors will attempt to present the opportunities for practical application of this method in cardiology based on the experience of renowned medical centers and first experience of the authors themselves. PMID- 12638331 TI - [Role of computed tomography in diagnosis of the vascular system]. AB - CT Angiography (CTA) is becoming a popular method used in the diagnosis of the vascular system. The authors present own experience acquired from 163 CTA examinations. In 98 cases, the CT angiography images were the elprimary indications for surgery or intervention procedures. Follow-up CTA examinations were performed in 65 of the cases. CT angiography is a reliable method in vascular diagnostics. Further progress in the development of this technique will lead to the replacement conventional angiography in most clinical diagnostic indications. PMID- 12638330 TI - [Evaluation of atherosclerosis progression in patients treated repeatedly with hemodialysis]. AB - The main cause of increased morbidity and mortality in patients on maintenance hemodialysis are cardiovascular complications due to accelerated atherosclerosis. Lipid profile disturbances, increased levels of homocysteine (HC), fibrinogen, iPTH as and Ca-P abnormalities seem to be the important factors in atherosclerosis progression. The aim of the study was to evaluate the advancement of atherosclerosis in patients on maintenance hemodialysis and to assess levels of pro-atherosclerotic factors. The study included 50 patients (30 M, 20 F) aged 21-75 years (mean 48.6 y) hemodialyzed three times a week for 12 to 282 months (mean 114.5 m). The homocysteine, fibrinogen, iPTH, calcium, phosphate and indices of lipid metabolism such as total cholesterol, HDL, LDL, triglicerides, Lp (a) and Apo B were measured before hemodialysis. To evaluate the advancement of atherosclerosis, all patients underwent MSCT using Somatom plus 4 apparatus with calcium scoring (CS) calculation and B-mode ultrasound for IMT using Acuson 128 XP 10 apparatus. The above applied methods helped to evaluate changes in central and peripheral arteries. In CS testing, coronary artery calcifications were present in 36 patients (72%). The CS ranged from 0 to 4345, with the mean CS being 584 (SD = 1012). The CS correlated significantly with age (r = 0.39; p < 0.005), P (r = 0.33; p < 0.05), CaxP product (r = 0.39; p < 0.05), iPTH (r = 0.43; p < 0.001) and with IMT (r = 0.56; p < 0.0001). The IMT ranged from 0.5 to 1.5, with mean of 0.89 (SD = 0.28). The IMT correlated significantly with age (r = 0.54; p < 0.0001), time on dialysis (r = 0.40; p < 0.01), fibrinogen (r = 0.43; p < 0.02), LDL (r = 0.30; p < 0.05), P (r = 0.29; p < 0.05), and CaxP product (r = 0.3; p < 0.05). Based on our study, we conclude that age, time on dialysis, increased levels of homocysteine, LDL cholesterol, fibrinogen, P, and iPTH as well as Ca-P disturbances are strong predictors of atherosclerosis in HD patients. PMID- 12638333 TI - [Evaluation of myocardial viability using magnetic resonance imaging--case report]. AB - The authors described the usefulness of magnetic resonance imaging in the evaluation of the viability of the left ventricular myocardium in a patient with a history of cardiac infarction treated with primary coronary angioplasty (primary PCI) and with symptoms of postreperfusion injury of the left ventricular muscle (no-reflow phenomenon). The perfusion of the left ventricular myocardium was evaluated with the use of paramagnetic contrast. The heart movement was evaluated at rest and after the administration of a small dose of dobutamine. The use of magnetic resonance tomography made it possible to pinpoint the area of stunned and necrotic myocardium. PMID- 12638332 TI - [Evaluation of patency of coronary artery bypass grafts and stents using multislice spiral computed tomography in comparison with angiography]. AB - The aim of the study was to assess usefulness of multislice spiral computed tomography--MSCT (Somatom Plus 4 Volume Zoom, Siemens) in non-invasive assessment of the potency of coronary artery bypass grafts both venous and arterial grafts as well as stents. METHODS: The study was performed using the MSCT method with Heart View Software with retrospective electrocardiographic gating. Nonionic, hypo-osmolar contract media was infused into a peripheral vein to highlight the coronary arteries and bypasses. All patients, due to clinical indications, underwent coronary and bypass angiography. MATERIAL: Two group of patients (82 persons) in total were evaluated: after aorto-coronary bypassing (Group I--57 persons, incl. 40 males and 17 females) and after stent implantation (Group II- 25 persons, incl. 19 males and 6 females). The patients were referred to invasive diagnostics for recurrence and aggravation of clinical symptoms. RESULTS: In group I, the patency of 187 bypasses was evaluated (21 arterial and 166 venous). In MSCT, was found in 131 of the bypasses patency (114 venous and 4 arterial). 56 bypasses, (52 venous and 4 arterial) were obstructed. In the bypassography performed were obstructed 53 (5 venous and 3 arterial), 134 bypasses were patent (116 venous and 18 arterial). In the bypass patency assessment using MSCT compared to coronarography, specificity of 94.7% and sensitivity of 92.4% were achieved. In group II, patency was evaluated in 26 stents: 21 implanted to native vessels and 5 in venous bypasses. 19 stents in native vessels were found patent, whereas 2 were obstructed. In the coronary bypasses performed, patency was confirmed in 19 stents and 2 were obstructed. In MST obstruction was demonstrated in 4 stents in coronary bypasses, which was confirmed in bypassography. One stent was assessed patient in both methods. CONCLUSION: The MSCT method is useful for assessment of patency of venous and arterial bypasses as well as stents. PMID- 12638335 TI - [Effect of cardiac rehabilitation on left ventricular function and exertion capacity in patients with chronic congestive heart failure]. AB - Cardiac rehabilitation is proven to be an effective method of treatment in congestive heart failure (CHF) and it has a positive influence on exercise capacity. There are controversies concerning the influence of physical rehabilitation on morphological and functional parameters of the left ventricle. Cardiac magnetic resonance (CMR) is a very precise noninvasive method of cardiac examination. The aim of this study was to assess changes in exercise capacity and in morphological parameters of the left ventricle in patients with CHF (NYHA class II and III, ejection fraction less than 35%) undergoing 6-months of physical rehabilitation. 16 patients were divided into two groups: A (7 pts) trained with progressive workload and group B (9 pts) did not trained. Groups were not different in terms of age, gender and NYHA class. In all patients, CMR and cardiopulmonary exercise test were performed at baseline and after 6 months. RESULTS: At baseline, both groups were similar with respect to the LV dimensions, thickness of LV walls and exercise capacity. At follow up, we observed an increase in exercise capacity in only the trained group (A). Exercise time increased from 357.1 +/- 30 sec. to 830.1 +/- 59 sec. (p < 0.05); max. workload from 2.7 +/- 0.36 MET to 6.21 +/- 0.8 MET (p < 0.05) and VO2 peak from 14.3 +/- 1.8 ml/kg/min to 18.8 +/- 2.1 ml/kg/min (p < 0.05). In group B (not trained) there were no changes after 6 months in these parameters. Morphological LV parameters were similar in both groups at baseline and after 6-months of rehabilitation. PMID- 12638334 TI - [Usefulness of magnetic resonance imaging in diagnosis of mitral valve anulus abscess--case report]. AB - Magnetic resonance imaging (MRI) is a non-invasive method characterised by high temporal and spacial resolution that makes it possible to obtain very high quality pictures. It is a less invasive method than TEE and is very significant in the diagnosis of heart tumours. MRI makes it possible to assess the parameters influencing hemodynamic as well as morphological qualities of the tumour such a its size, its movement, its relation to the surrounding structures and the presence of a capsule. With the use of additional programming, it is possible to evaluate, indirectly, the metabolism of the diagnosed lesion as well as the degree of blood perfusion. Thus, MRI diagnosis of the heart is a valuable complementary technique in the verification of the diagnosis and in the referral to cardiosurgical treatment. The aim of this paper is to present the use of MRI in the diagnosis of an abscess of the mitral valve anulus in a female patient who did not agree to the transoesophageal echocardiography (TEE). In this case, the usefulness of MRI to establish the diagnosis of the anular abscess of the mitral valve was proved. The final diagnosis was confirmed during the operation. PMID- 12638336 TI - Clinical application of transthoracic Doppler echocardiography to assess coronary flow reserve. AB - This review discusses methods (focusing on noninvasive transthoracic Doppler echocardiography) to measure coronary endothelium-independent and endothelium dependent vasodilatation. Coronary flow reserve is an important functional parameter to understand the pathophysiology of coronary circulation. Coronary flow reserve measurement is used to assess epicardial coronary stenoses or to examine the integrity of microvascular circulation. In the presence of coronary artery stenosis, coronary flow reserve is an important hemodynamic parameter for clinical decision making (for example to refer or defer patients with intermediate stenosis [40-70%] to PTCA). In the absence of stenosis in the epicardial coronary artery, the coronary flow reserve may be decreased when coronary microvascular circulation is impaired by arterial hypertension with or without left ventricular hypertrophy, diabetes mellitus, hypercholesterolemia, hypertrophic cardiomyopathy, aortic stenosis or syndrome X. PMID- 12638337 TI - [Usefulness of two-dimensional echocardiography for evaluation of left ventricular aneurysm. Comparison with magnetic resonance imaging and intraoperative evaluation]. AB - Aim of the study was to validate 2D echocardiography (2DE) for quantifying left ventricular aneurysm (LVA). Assessment if the initial parameters of LVA size could predict the choice of surgical technique. MATERIAL AND METHOD: Group 1 consisted of 10 patients with LVA who underwent 2DE and magnetic resonance imaging (MRI) as a reference method. Apical 4-chamber plane was used in both techniques to determine LVA area and volume (area-length method). Group 2 consisted of 33 patients with LVA who underwent surgical endoventriculoplasty (EVP): 18 with patch (P), 15 without patch. 2DE was performed before and after surgical procedure. LVA-area, LVA-volume were determined. RESULTS: Group 1 (2DE/MRI): Mean LVA-area and LVA-volume in 2DE were 17.5 +/- 6 cm2 and 62.6 +/- 32 ml, respectively, and in MR, they were 20.9 +/- 7 cm2 and 65.5 +/- 35 ml, respectively. Excellent correlation was found between 2DE and MRI: r = 0.85 for LVA-area and r = 0.81 for LVA volume (p < 0.001). 2DE underestimated LVA measurements in comparison with MRI results. Group 2 (surgical patients): Mean LVA-area was 15 cm2, LVA-volume 50 ml. LVA exceeding 50 ml was considered as large. 2DE LVA size estimation was in agreement with surgical assessment. All but one patient with LVA-volume surpassing 56 ml were operated on with P. EVP without P was done for smaller LVA. CONCLUSION: 1. With MRI as a reference method, 2DE can accurately quantify LVA size providing information about disturbances in LV geometry. 2. 2DE parameters of LVA-area and LVA-volume are valuable in planning operative method of EVP. PMID- 12638338 TI - [Rapid resolution of ST segment elevation predicts recovery of left myocardial contraction in patients with acute myocardial infarction treated with percutaneous coronary angioplasty]. AB - BACKGROUND: Early epicardial vessel patency and tissue perfusion limit infarct size, improve survival and are crucial for optimum functional recovery of the ischaemic myocardium in patients with acute myocardial infarction (AMI). Coronary angiography has been considered the "gold standard" in assessment of reperfusion. Restoration of coronary patency is not a guarantee of myocardial cellular perfusion. ST-segment elevation resolution observed in electrocardiogram (ECG) early after initiation of primary PTCA could be potentially correlated with reperfusion. AIM: The aim of the study was the prospective assessment of left ventricular (LV) contractility recovery in patients after primary PTCA for AMI in 3 and 6 month follow-up according to early ST-segment elevation resolution analysis. METHODS: 111 consecutive patients (80 men and 31 women age 36-78) with AMI treated by successful primary PTCA (Thrombolysis in Myocardial Infarction grade 3 flow and residual stenosis less than 30%) within 12 h of chest pain onset were enrolled in the study. 12-lead ECG was performed on admission and 1 hour after the beginning of the PTCA. The patients were divided into 2 groups according to ST-segment elevation resolution in the ECG (> 50% depression of the elevated ST-segment) within 1 hour after the procedure. The mean pain-to-balloon time was not significantly different in both groups. In each patient, global ejection fraction (EF) and wall motion index (WMI) were derived immediately after primary PTCA and after 3 and 6 month follow-up. RESULTS: Of the 111 patients studied, 82 (68%) had early ST-segment resolution and 29 (32%) did not. Contractility disturbances of infarct related ventricular wall, evaluated by EF and WMI, were significantly higher in the group of patients without early ST segment resolution immediately after primary PTCA. EF and WMI improved significantly after 3 and 6 months in the group of patients with improvement was insignificant ST-segment elevation resolution, while in the group without ST segment resolution. CONCLUSIONS: 1. Presence of early ST segment elevation resolution after angiographically successful primary PTCA identifies patients who are more likely to benefit from the early restoration of flow in the infarct related artery. 2. TIMI measures greatly overestimate the success of primary PTCA; they only assess vessel patency, not myocardial cellular perfusion. 3. "Electrocardiographic reperfusion" provides a real-time physiologic marker of cellular perfusion and is a significant predictor of LV contractility recovery- more useful than angiographic reperfusion. 4. ST-segment monitoring is a reliable, non-invasive and inexpensive method to evaluate myocardial perfusion. PMID- 12638339 TI - The contribution of non-invasive imaging modalities to the diagnosis of left ventricular pseudoaneurysm. AB - BACKGROUND: Left ventricular pseudoaneurysm (LVPA) is a rare entity characterized by a tendency to spontaneous rupture due to its morphology, a lack of myocardial fibers and fibrous tissue delineating the cavity. An early diagnosis is essential in order to guide appropriate therapy. PURPOSE: To determine the diagnostic accuracy of different imaging techniques, treatment results, and prognosis of patients (pts) with LVPA. METHODS: We evaluated the incidence of LVPA during a five-year period. The initial clinical presentation, the etiology of LVPA, time between symptom onset and diagnosis, and use of various non-invasive techniques were studied. Mean follow-up was 15 months. RESULTS: Of 19113 pts admitted to our Institute in a five-year period, LVPA was diagnosed in 11 pts (0.05%) (mean age 51 +/- 3.9 years, 8 men). The diagnosis of LVPA was confirmed by surgery in 4 pts, and by pathology in 2 pts. LVPA was an incidental finding in one asymptomatic pt, it was diagnosed in 6 pts presenting with an acute myocardial infarction (AMI) and in 4 pts presenting with LV failure. The main etiology was coronary artery disease (CAD) (9 pts), with the remaining 2 cases being post traumatic (thoracic stab wound, surgery). LVPA location was postero-inferior in 6 patients, infero-lateral in 3 patients, and anterior in 2 patients. ECG, X-ray and TTE were performed in all cases. 6 pts had a radionuclide angiography (RNA), 3 pts had a computed tomography (CT) scan and 2 pts had a magnetic resonance imaging (MRI) study. Two-dimensional transthoracic echocardiography (TTE) provided information regarding LVPA dimensions and LV-LVPA flow. Four pts were operated (one died). Of the seven non-operated pts., 5 died. CONCLUSIONS: The clinical presentation was variable and non-specific. The most frequent cause of LVPA was MI and the most frequent location was inferior. Echocardiography offered the most reliable information when compared to ventriculography. Because clinical examination, ECG, X-ray data are non-specific for the diagnosis of LVPA, an adequate TTE study performed with a high clinical index of suspicion (especially in pts with inferior MI or thoracic trauma) could facilitate the early diagnosis of LVPA and could be relevant to outcome. PMID- 12638341 TI - Clinical application of 3-dimensional echocardiography. AB - Three-dimensional echocardiography has emerged as a novel non-invasive imaging modality enabling the registration and analysis of volumetric datasets containing information on cardiac structure. This review describes the principles of the method, different acquisition modes and the display options in perspective of clinical applications. Emerging clinical applications are discussed, including improved quantification of cardiac volumes and the ability of more complete, versatile and intuitional display of cardiac pathology. According to the available literature, the most probable fields for practical use of three dimensional echocardiography are cardiac interventions in such conditions as valvular disease, congenital pathology (septal defects) or cardiac tumors. The most expected development is the introduction of fast, user-friendly systems allowing real-time data acquisition and data processing. PMID- 12638340 TI - [Predictive value of pharmacologic echocardiographic test for early and late cardiovascular events]. AB - PURPOSE: Stress echocardiography (SE) and radionuclide perfusion imaging are competing diagnostic methods in patients with ischemic heart disease. The prognostic value of nuclear studies has been much better established so far. The purpose of the study was to evaluate the long term prognostic value of pharmacological stress echocardiography. METHODS: We followed up 224 patients (62% men, aged 55 +/- 10 yrs) diagnosed with pharmacological stress echocardiography with ischemic heart disease, 46% with angina, 39% with hypertension, 43% with hypercholesterolemia, 12% current smokers, 44% previous smokers, 10% with diabetes mellitus. Positive SE result was defined as new or increased wall motion abnormalities. Follow-up period ranged from 12 to 91 months, average 53 +/- 19 months. Myocardial infarction, recurrent unstable angina requiring hospitalization and need for invasive surgical revascularization or death were defined as adverse outcome. RESULTS: Positive SE result was observed in 109 patients (48%). During follow-up, 21 patients died (9%) and in 132 patients (59%) other cardiac events were observed. Positive predictive value (PPV) during follow-up was 51% (for cardiac death--8%) for the first year and 7 12% (for cardiac death--0-4%) in the years 2-5. 51% for first year and 7-12% for 2-4 year. Negative predictive value (NPV) was 82% for the first year (for cardiac death--98%) and 79-94% (for cardiac death--99-100%) in years 2-5. 82% for first year and 94-92% in further follow-up. However, 1-year PPV for mortality was 8% with a high NPV (98%). 5-year death-related NPV of the test is very high (100%). CONCLUSION: Negative result of SE has a moderate NPV for 1 year cardiac event but high NPV for mortality. NPV in 2-5 years of follow-up in patients with negative SE results is high. PPV is higher in the first 1 year than in further follow-up which results partly from selection bias and remains stable thought area 5 years. PMID- 12638342 TI - [Evaluation of left ventricular diastolic function with tissue doppler echocardiography (TDI) in patients after angioplasty of the artery responsible for infarction]. AB - To assess the ability of left ventricular (LV) diastolic function to improve following angioplasty of an infarct related artery (IRA), 39 patients were evaluated with the Tissue Doppler Imaging (TDI). Studied group comprised of 30 males and 9 females (age 53.3 +/- 8.3 yrs), who were 1 to 6 months after myocardial infarction (MI). They qualified for angioplasty of IRA if dobutamine stress echocardiography proved viability of the affected myocardium. Regional wall diastolic function was assessed by TDI 1 day before (exam 1), 1-3 days (exam 2) and 30 days (exam 3) after successful angioplasty. TDI myocardial diastolic velocities (Em and Am wave, Em/Am ratio) and time intervals (isovolumic relaxation time IVRT, rapid filling time RFT, atrial filling time AFT) were measured in the long axis of posterior (20 pts), anterior (17 pts) and lateral (2 pts) walls. Regional E wave velocity was increased between exam 1 and 3 (6.0 +/- 1.5 cm/s vs 6.7 +/- 1.7 cm/s, p < 0.05), the same as Em/Am ratio (1.0 +/- 0.4 vs 1.3 +/- 0.9, p < 0.05). There was no significant change in Am wave velocity. The regional RFT was prolonged between exam 1 and 2 (151 +/- 39 ms vs 170 +/- 30 ms, p < 0.01), whereas IVRT was shortened (103 +/- 21 ms vs 87 +/- 20 ms, p < 0.001). CONCLUSIONS: Regional diastolic function showed improvement as assessed by TDI after successful angioplasty of IRA. TDI allows the assessment of regional diastolic performance and its dynamics. PMID- 12638343 TI - Clinical application of 3-dimensional echocardiography--a 3-year single center experience. AB - 3-dimensional echocardiography (3DE) is a novel and rapidly evolving echocardiographic technique. Theoretically, 3DE should be helpful in difficult cases when 2-dimensional echocardiography (2DE) cannot deliver required information due to complex spatial relationships in diseased heart of inability to obtain a desired cross-sectional view. Accurate quantification is another potential benefit of 3DE. METHODS: In this study we present our 3-year experience in 3DE applied to selected clinical population. Retrospective analysis of our database provided data of 204 3DE studies performed with TomTec Echoscan 3.1 using rotational device from transthoracic or transesophageal window. RESULTS: A total of 474 3DE acquisitions were performed. The main indications for 3DE were: Assessment of left ventricular function (51);--valvular heart disease (47), assessment of thoracic aorta (36), congenital anomalies (36) and cardiac masses (22). Quality of resulting 3D image was graded as 0--bad, 1--satisfactory, 2- good, 3--demo. Average quality of reconstruction was 2.1 +/- 0.8, with 73% of interpretable (score 2-3) studies. Compared to 2DE, additional morphologic or quantitative information was obtained in 47% of patients, including: left ventricular function assessment (41%); valvular heart disease (40%); assessment of descending aorta assessment (31%); congenital anomalies (64%); and cardiac masses (64%). CONCLUSION: 3DE can be incorporated into the routine diagnostic pathway with a high feasibility rate. 3DE is a valuable tool in clinically indicated cases by providing additional qualitative or quantitative information in 47% of cases. PMID- 12638344 TI - [Assessment of myocardial perfusion using myocardial contrast echocardiography- case report]. AB - A 58-year-old woman with symptoms of angina underwent coronary angioplasty, which showed critical narrowing of the right coronary artery (80%) and borderline narrowing of the left anterior descending coronary artery (50%). We decided to analyse myocardial perfusion in this patient to estimate the degree of atherosclerosis in the target arteries. She underwent myocardial contrast echocardiography at rest and during dobutamine stress. The examination showed a transient perfusion defect in the right coronary bed and in the left anterior descending coronary. A similar perfusion defect was revealed in scintigraphy. We carried out a successful two vessel percutaneous coronary revascularisation. PMID- 12638345 TI - [Stress echocardiography with dobutamine in patients with aortic stenosis]. AB - Echocardiographic examination at rest is usually an adequate diagnostic method in evaluating patients with aortic stenosis and in assessing the severity of stenosis. However, in patients with aortic stenosis with coexisting coronary heart disease and in patients with poor left ventricular systolic function and low gradient aortic stenosis, dobutamine stress echocardiography should be performed. This test is helpful in differentiating between significant fixed aortic stenosis with secondary left ventricular dysfunction and severe left ventricular dysfunction coexisting with nonsignificant aortic valve stenosis. This examination is necessary to select the proper method of treatment and is helpful in risk stratification. PMID- 12638346 TI - [Safety of stress echocardiography with dobutamine used in patients with aortic stenosis and left ventricular dysfunction]. AB - THE AIMS: The aim of the study was to assess the safety of dobutamine echocardiography (DE) in patients with aortic stenosis and left ventricular ejection fraction < or = 45%. METHODS: 162 patients (mean age 59 +/- 13, 64% males) with resting transvalvular peak gradient < 70 mmHg underwent standard DE (doses 5-40 micrograms/kg/min) in a multicenter study involving 10 centers from Poland and Hungary. This analysis involves a subset of 39 pts (34 M, 5 F, age 59 +/- 13) identified according to the criterion of EF < or = 45% and mean aortic gradient < or = 35 mmHg. Clinically indicated coronary arteriography was performed in all patients with left ventricular dysfunction. RESULTS: Peak dobutamine dose was 29 +/- 10 micrograms/kg/min. Peak heart rate was 112 +/- 26 bpm. Systolic arterial pressure was 148 +/- 20 mmHg and diastolic pressure 86 +/- 14 mmHg. Transaortic mean gradient and aortic valve area increased 35 +/- 34% and 19 +/- 15% during peak dobutamine dose, respectively. Ejection fraction improved 16 +/- 14%. The test was positive in 17 (43.6%) patients, negative in 11 (28.2%) patients and non diagnostic in 11 (28.2%) patients. There were no deaths, myocardial infarctions or episodes of sustained ventricular tachycardia as a result of DE. The test was terminated when the following conditions were met: Target heart rate (23.%). Left ventricular asynergy (46.2%). Maximal established dose achieved (7.7%). Side effects (20.5%). The most common side effects were ventricular arrhythmias (35.9%), dyspnea (20.5%) and hypotension < 20 mmHg (10.3%). CONCLUSION: Despite frequent side effects, DE can be safely performed in patients with aortic stenosis and depressed left ventricular contractility. PMID- 12638348 TI - [Comparative evaluation of 2 epidemiologic surveillance systems of hepatitis A]. AB - Epidemiological surveillance represents an important tool for the prevention of hepatitis A. In Italy, the surveillance of hepatitis A is based on the routine notification system of infectious diseases (from 1996 it was replaced by SIMI, Computerised System of Infectious Diseases) and on SEIEVA (Epidemiological Integrated System of Acute Viral Hepatitis). From 1997 to 2000 the two surveillance systems have been compared in Puglia region to evaluate match rate, completeness and sensitivity. A total of 7.018 records from SIMI and 4.413 from SEIEVA have been analyzed. During this period, the 37.0% of cases notified to SIMI match SEIEVA records whereas the 58.8% of cases reported to SEIEVA match cases notified to SIMI. Therefore, the match rate between the two systems was 29.0%. The sensitivity of SIMI and SEIEVA resulted 79.4% and 49.9% respectively. The comparison of the two surveillance systems allowed to recognise problems in local organisation. Therefore, a more adequate training should be supplied to workers involved in the surveillance activities. PMID- 12638347 TI - [Implants with aortic valve homografts in patients with critical left aortic stenosis]. AB - This case study presents two patients with aortic stenosis and infective endocarditis (IE) who underwent homograft aortic valve replacement. The first patient is a 67 year old practicing surgeon. Twenty one years ago, he was admitted to our department for critical aortic stenosis, infective endocarditis (IE) and heart failure. He underwent homograft aortic valve replacement. After the surgery his condition improved dramatically and he returned to his practice. Current echocardiographic study shows normal function of the left ventricle and normal gradient across homograft aortic valve. The next case, a 33-year old happy mother of 5 children, is particularly interesting. When she was 15 years old, she was referred to surgery, diagnosed with bicuspid aortic valve stenosis, subaortic muscular stenosis and aneurysm of aortic sinus of Valsalva. She underwent homograft aortic valve and root replacement and excision of the sub-aortic muscle bulge. Eleven months later, she required another operation due to active IE. The St. Jude Medical aortic valve was implanted. Two months after the surgery a dysfunction of the implanted artificial valve was diagnosed. She again underwent the homograft aortic valve and root replacement with good long-term results. In the period spanning 1987-1997, she managed to deliver five babies without any complications whatsoever. Seventeen years later, the homograft aortic valve is still functioning fairly well. The homograft aortic valve replacement may be regarded as a viable option for patients with aortic stenosis and IE and for young women suffering from aortic valve disease who intend to be prospective mothers. PMID- 12638349 TI - [Sexual knowledge, attitudes, and behavior in HIV-seropositive individuals at the AIDS referral center Latina]. AB - AIM: The objective of the study was to investigate the knowledge, attitudes and sexual behaviours of HIV positive users of the AIDS Centre of Latina, with the scope of enabling one to program effective interventions on the theme of sex education. METHOD: An anonymous self-administered questionnaire containing questions about the socio-demographic characteristics of the subject, about the transmission of HIV, about sexual behaviours and about the relationship between sexual habit and being HIV positive, was compiled in the period between 1st October 2000 and 3rd January 2001. In all, 259 subjects (92.84% of eligible subjects), of which 166 males and 93 females aged between 20 and 62 years (mean = 38.51), participated in the study. RESULTS: Regarding the knowledge about HIV/AIDS, 63.7% believed that a vaccine against AIDS is currently unavailable. Nearly all responders thought that the HIV virus can be transmitted through the shared use of infected syringes among drug addicts, through homosexual and heterosexual intercourse and through blood transfusions with infected blood. Vertical transmission, during pregnancy, is considered possible by 78.6% of respondents. Regarding the sexual behaviours of participants, 81.9% reported to have full intercourse, of which 78% declared to use condoms during sexual intercourse. As regards the attitudes about sexuality and HIV, 83.7% reported that education is a fundamental factor in the reduction of risk behaviours, though 32.9% considered it embarrassing to ask their partner to use condoms and, surprisingly, 22.3% would consent to unprotected sex even if their partner wasn't wearing a condom. CONCLUSION: In the area of STDs, among which HIV and AIDS, it is worrying that many people overlook the most elementary forms of prevention. This study confirms the necessity for an adequate form of informative counselling for those who are HIV positive. PMID- 12638350 TI - Is diabetes mellitus a risk factor for HCV infection? AB - The aim of this study was to investigate whether or not the diabetes mellitus may be considered a risk factor for the HCV infection. The HCV seroprevalence was evaluated in 254 diabetic subjects, whose anamnestic data and risk factors are known, in comparison to 223 first-time blood donors, carefully age- and gender matched. The statistical analysis showed that the studied groups belonged to the same population (Mann-Whitney U test) and that there were no significant differences between cases and controls as regards HCV prevalence (Yates corrected chi 2 test). The obtained data underline the importance of the control group selection, especially in the studies considering age-related pathologies. The authors disprove type 2 diabetes as a risk factor for the HCV infection and consider that this is a valid hypothesis only when the hepatitis C was unknown and not adequate prevention was used. PMID- 12638351 TI - [Microbiological study of soft cheese purchased in small and middle-sized distribution networks]. AB - The aim of the work is the evaluation of the microbial content of soft cheese when it comes on the table of the consumer. To reach this purpose we have bought, as simple clients, the product in the small and average distributions that apply the HACCP (Hazard Analysis Critical Control Point) plain and in the small distributions which don't apply it, the collecting and the microbiological analysis of the product has been performed on the different pieces that belong to the whole form of cheese during the days of the week, observing a progressive increase of the bacterial number; besides we have found in the bought cheese that when the form of cheese was at the end, values overcame the limits of law. The increase of the bacterial numbers was greater in the products purchased in the average distributions in order with the application of the self-control, and in the small ones that don't effect this type of selling. The deduction was that the HACCP system has not been correctly applied in all the shops tested and in the exercises without the HACCP plain the contamination levels are higher. The health risk connected with not correct manipulation by the employees still results very elevated. PMID- 12638352 TI - [Evaluation of the Surveillance System of Acute Flaccid Paralysis in Puglia: 5 years of work]. AB - Surveillance of acute flaccid paralysis (AFP) is the milestone to monitor the progress toward poliomyelitis eradication aim, fixed by WHA in 1988. Active AFP surveillance started in Apulia in 1997; this work evaluates five-year period activities. In this period, the total number of cases notified was 48, 7 of which were resident out of Apulia. Twenty-five were males and 23 females; the age ranged between 1 month and 15 years. Any collected serum specimens showed protective antibody levels against polioviruses. Polioviruses type 1 and type 2 Sabin-like were isolated from stool samples collected from two AFP patients. AFP surveillance targets improved in the years, with only exception, in 2001, of second serum specimen collected within 14 days because of children were discharged earlier form the hospitals. Apulia experience demonstrates the achievement of good levels of AFP surveillance targets. System sensitivity has been optimal in 2001 with a number of notified cases threefold the expected value and adequate specimen sampling (80%). Additional involved hospitals and availability of increased and dedicated human resources contributed to this outcome. The effort to achieve WHO targets for AFP surveillance needs to be maintained in next years until global certification of eradication will be declared. PMID- 12638353 TI - Epidemiology and prevention of domestic injuries among children in the Verona area (north-east Italy). AB - In the developed world, domestic injuries (DI) are an important cause of morbidity, temporary or permanent disability, and death in early life, the social and economic costs of which are often underestimated. To assess the epidemiology of this phenomenon in an area of north-eastern Italy, a retrospective investigation was performed with an anonymous questionnaire administered to the parents of approximately 3000 children aged between 3 and 15 years. More than 45% of the sample had suffered at least one DI in their lifetime. The most common involved falling, wounding and scalding, and particularly affected children above 4 years old. The various types of injury were analyzed and correlated with the parents' personal parameters, the circumstances and the location of the accident. The type of aid required (medical advice was sought in more than 70% of cases) and the outcome of the DI (26% cases of temporary disability and 2% of permanent disability) were also assessed. The majority of DI could be prevented by a capillary campaign on the prevention of domestic hazards in childhood, preferably as part of a holistic approach to the problem that also considers their living conditions in architectural and interior design terms. PMID- 12638354 TI - [Evaluation of the efficacy of the "Spazi Giovani" of family clinics. Results of 3 ASL services of the City of Milan]. AB - "Spazio Giovani" is a service offered once a week by many Family Counselling Services and aimed at addressing the needs of tenagers and young people who can access it free of charge and without booking an appointment. This kind of services started in the Eighties to address the needs of young people who were sexually active and often not accessing conventional services. The objective of this study has been to assess how attitudes for unprotected sex and at risk sex behaviours decrease in a population attending the Youth Corner on a regular basis in comparison with the same attitudes and behaviours in a group of first-time users of the service over a six month period. The research, involving 301 teen and young females showed significant differences in protected behaviours between the two groups. No significant differences in attitudes were put in evidence by the study. PMID- 12638355 TI - [Quality of health care services as perceived by the customers: issues of method or context?]. AB - Customer satisfaction is considered an important indicator of the quality of care. Its definition as well as the identification of the variables which affect it, rise many cultural and methodological issues. In order to give a contribution to the debate on such topics, we compared the patients' satisfaction detected before and after the transferral of the San Salvatore Hospital of L'Aquila to new and functional structures. The comparison aimed at evaluating the methodological and cultural entailments involved in customers satisfaction surveys, which focus the improvements in terms of health care as well as variation of satisfaction. The presence of contradictory elements in the expression of the satisfaction referred to the technical and informative aspects, seems to indicate that patients can express an high satisfaction degree independently from the real professional and technical quality performed. Such evidences, that anyway must be interpreted according with the methodological cautions of a non validated questionnaire, should foster stronger efforts in promoting sanitary education of the customers, devoted to the specific rights involved, as well as in making use of rigorous methodologies to detect the phenomenon. PMID- 12638356 TI - [Legislative Decree 25 of 2/2/2002: prevention of chemical agents risks]. AB - The Authors, with the present contribution, illustrate and comment the recent legislation aimed to protect workers from risks due to chemical agents and compare it with the previous legislation. A careful analysis makes it clear the urgent need to realize optimal safety standards for workers in the chemical industry, in order to protect them from avoidable risks. PMID- 12638357 TI - [Corrado Tommasi Crudeli and Vittorio Del Vecchio (pioneers in public health sciences)]. PMID- 12638358 TI - [The debate in Italy and in Europe about the adoption of pneumococcal vaccine in children]. AB - Since the introduction of the conjugate vaccine PnC-7, for the prevention of invasive pneumococcal infections for at-risk children, a controversy has arisen about the need for a universal vaccination under the age of two years. This article reviews the pathogenesis and the changing epidemiological pattern of pneumococcal diseases, the emergence of drug resistant S. pneumoniae and the costs and benefits of vaccination. Furthermore, the recommendations of various European countries and an up to date of the Italian ones are illustrated. It concludes with the public health perspective on the adoption of this pneumococcal vaccine and future recommendations for vaccination. PMID- 12638359 TI - [Epidemiology of meningitis, pneumonia and acute otitis due to Streptococcus pneumoniae in pediatric in Italy]. AB - Streptococcus pneumoniae is one of the most important bacterial pathogens in pediatrics, both from a quantitative and from a qualitative point of view. It can cause severe diseases such as occult bacteremia, sepsis, meningitis and it is also involved in the determination of a relevant part of very common illnesses such as pneumonia, acute otitis media and rhinosinusitis. However, most of the data regarding the epidemiology of pneumococcal infections in pediatrics have been collected in the U.S.A. and in other industrialized Countries different from Italy. In Italy only studies concerning meningitis and the carrier state of pneumococcus are at the moment available. They indicate that also in Italy S. pneumoniae seems to play a significant role in the determination of several pediatric diseases and that quite similar are the group of serotypes mainly involved in the determination of both invasive and not invasive diseases. PMID- 12638360 TI - [Surveillance of invasive diseases]. AB - Severe clinical conditions are due to invasive diseases that are not routinely reported in a routine surveillance system, in Italy. Nevertheless we can estimate the incidence of invasive diseases considering the surveillance system of bacterial meningitis, the Hospital Discharge Registry and the laboratory-based surveillance, this is useful to outline the trend of S. pneumoniae infections. We report the results of some Italian studies on the pneumococcal infections together with some international studies. PMID- 12638361 TI - [Efficacy of the heptavalent pneumococcal vaccine against meningitis, pneumonia and acute otitis media in pediatric age. Theoretical coverage offered by the heptavalent conjugate vaccine in Italy]. AB - All the clinical trials carried out to evaluate the heptavalent pneumococcal conjugate vaccine have clearly demonstrated that it is safe, well tolerated and, despite some differences in the different diseases, effective in the prevention of all the pneumococcal illnesses. Considering that the available data demonstrate that the serotypes involved in the determination of pneumococcal diseases are, in Italy, the same found in the U.S.A. where the vaccine has been formulated, it is reasonable to think that it will be very effective also in our country. We hope that this vaccine can be systematically prescribed also in Italy: this could permit to reduce the number of both common and severe pneumococcal diseases at the moment largely present among italian children. PMID- 12638362 TI - [7-valent conjugate Pneumococcal vaccine and nasopharyngeal cavity]. AB - 7-valent conjugate pneumococcal vaccine is effective for preventing invasive pneumococcal diseases in childhood. The present serotype formulation of the conjugate Pneumococcal Vaccine is effective for preventing 75-90% of the invasive pneumococcal disease cases, depending of the vaccine-serotypes distribution in the several countries. Recent studies, carried out in the north-american context, show that mass vaccination campaign by 7-valent conjugate Pneumococcal Vaccine in childhood, reduce, in vaccinated subjects, the nasopharyngeal colonization by vaccine-serotypes, that are much more joined to antibacterial resistance. This implies non only an active and specific protection for vaccinated, but also for family and community contacts, children and adults. This interesting results, need to be confirmed also in european countries, open a new view for a wider pharmacoeconomic evaluation of the 7-valent conjugate Pneumococcal Vaccine in childhood. PMID- 12638363 TI - ["Herd immunity" after administration of Pneumococcal conjugate vaccine and its impact on vaccination campaigns]. AB - Parallel to the diffusion of conjugated pneumococcal vaccine in children, several studies have shown a reduction of the incidence of invasive pneumococcal disease both in the vaccinated and in the unvaccinated. Such an observation is not a surprise if one considers that a similar phenomenon has taken place also after the mass introduction of Hib vaccine, at its turn a conjugated polysaccharide vaccine. Another observation is that also the carriage of vaccinal pneumococci was reduced in both the vaccinated and the unvaccinated, while in some cases the carriage of non-vaccinal serotypes, definitely less pathogenic, could be increased. PMID- 12638364 TI - [Costs of pneumococcal infections and cost evaluation of heptavalent conjugate vaccine]. AB - The results of six economic evaluations on heptavalent conjugate pneumococcal vaccination in children are discussed. Although the data of most studies come from estimates, the last study from US, in which data are based on a large randomized trial, shows encouraging results. Similar results are obtained in an economic evaluation performed in Germany, although the study is based on estimated data. The paper concludes stressing the need to improve epidemiological, clinical and economical knowledge on the pneumococcal diseases, in order to identify the best cost-effectiveness strategy to prevent a diseases so important for children and the elderly. PMID- 12638365 TI - [Why are students lonely? Factors influencing loneliness in the school context]. AB - A significant amount of children and adolescents consider themselves as lonely because their essential social needs are not sufficiently met. The present study analyzes the contributions of group acceptance reflected by the social status, of social behaviors, reciprocal friendships, and family climate to the frequency and intensity of loneliness. About 1200 students of 6th and 8th grade of secondary school in Vienna were asked about their social experiences in school. Peers filled up questionnaires about the behavior of these students. A major part of the 11- and 13-year-old students had negative social experiences in school and about 15% were suffering from these experiences and considered themselves as lonely. Lack of acceptance in the group contributed to this self appraisal as well as a lack of friends and (according to the students) little support by the family. Social behavior of lonely students was remarkable for frequent victimizations and a low level of prosocial behavior. The impact of poor group acceptance on the feeling of loneliness was transmitted by these two features of social behavior. PMID- 12638366 TI - [Self esteem of chronically ill children and adolescence exemplified by obesity and congenital heart defect]. AB - Coping with a chronic illness challenges children and adolescents in addition to their normal developmental tasks. This double challenge probably endangers the development of a stable self-esteem. The present investigation explores the possibility whether these processes are different with respect to the kind of illness. Chronic illnesses such as obesity and congenital heart defects (CHD) serve as examples in comparing two samples (8-16 years): obesity (N = 54) as visible and partly controllable illness (with respect to the course of illness) vs. congenital heart disease (N = 56) as invisible and uncontrollable illness (with respect to the origin and course of illness). Self-esteem is measured by a scale (ALS) which focuses on the public areas "school" and "leisure time" and the private area "family". Children and adolescents with CHD (especially females) display an above-average positive self-esteem in all areas. Children and adolescents with obesity mainly display an average self-esteem, the females scoring above-average for the private area "family", the males scoring below average for the public area "leisure time". Furthermore, leisure-time related self-esteem is significantly lower for obese than for CHD subjects. These specific relations implicate differential accentuations for intervention programs. PMID- 12638367 TI - [Play therapy--psychotherapy with play as the medium: I. General introduction, psychoanalytic and client-centered approaches]. AB - Play therapies are psychotherapies with the medium of play primarily for children under 12 years of age, which can be differentiated according to their theoretical constructs and actual practice. Play therapies have gained importance and relevance in the 1990's, reflected in a wide range of publications. Following trends can be discerned: narrow concepts defined by individual schools of psychotherapy have been left. Different forms of play therapy, as well as behavioural and family therapy have been integrated. Focussed short-term and therapies for specific disorders have been developed. The aim of the first part of this paper is to present an overview of traditional forms of playtherapy, with a focus on the Individual Therapy of A. Adler, the Analytic Psychotherapy of C. G. Jung, Sandplay Therapy of D. Kalff and child-centered (non-directive) play therapy. PMID- 12638368 TI - [Disclosure of incest as an emancipatory act--once again: "The Feast" by Thomas Vinterberg]. AB - The author examines the Danish film "The Feast" referring to the dynamics of a typical incest family. The disclosure of the incest is the central point of the scenario of the film. It is understood as an expression of emancipatory strivings of the victim, the need to separate from denying collaboration and from identification with the role of the victim (identification with the aggressor). Closely connected with this emancipatory process is the development of the siblings out of their mutual relationships and those to their parents towards a self defined life. PMID- 12638369 TI - Reconsidering models of patient satisfaction and behavioral intentions. AB - This article considers several models of how patients integrate their reactions to hospital attributes and how these reactions impact their overall satisfaction and behavioral intentions. It finds that patients combine their reactions to the attributes by means of noncompensatory and nonlinear models to form their overall satisfaction or behavioral intentions. PMID- 12638370 TI - Commentary--patient satisfaction: what is the point? PMID- 12638371 TI - Corporate management of quality in employee health plans. AB - As large companies move their employees into managed care, they must concern themselves with the quality and price of their employees' health care. Based on a survey of Fortune 500 companies, we show that most are integrating several aspects of quality into their purchasing and contracting decisions by focusing on three dimensions--customer service, network composition, and clinical quality. Companies focus on the customer service dimension while the medical community emphasizes clinical quality. PMID- 12638372 TI - Learning curves in health care. AB - This article explores the uses of learning curve theory in medicine. Though effective application of learning curve theory in health care can result in higher quality and lower cost, it is seldom methodically applied in clinical practice. Fundamental changes are necessary in the corporate culture of medicine in order to capitalize maximally on the benefits of learning. PMID- 12638373 TI - Merger as marriage: communication issues in postmerger integration. AB - Many health care organizations have found themselves involved in mergers, acquisitions, alliances, networks, and other forms of structural change in order to reduce costs, improve utilization and service breadth, increase market leverage, and reduce variation in demand. This article examines three levels of communication issues that interact with one another and impact the likelihood of successful postmerger integration: affect, discursive frame, and negotiating position. PMID- 12638374 TI - Organizational learning, diffusion of innovation, and international collaboration in telemedicine. AB - The authors analyze competing forces affecting the diffusion of telemedicine practices across organizations, potential learning effects from telemedicine practice, and their implications for the development of telemedicine-based networks. They also speculate on the learning, diffusion, and institutional effects that telemedical collaboration may trigger; five sets of propositions are advanced to explain these effects. PMID- 12638375 TI - "Virtual" health care organizations and the challenges of improving quality. AB - This article examines the challenges of improving health care quality continuously within and across "virtual" provider organizations such as independent practice associations and physician-hospital organizations. It draws on recent research and theory about interorganizational networks in other fields to develop recommendations for securing physicians' commitment to quality improvement strategies in today's health care environment. PMID- 12638377 TI - Basal cell and squamous cell carcinoma. AB - OBJECTIVES: To describe the clinical and histologic subtypes, pathophysiology, recognition, and treatment options for basal cell and squamous cell carcinoma, and the molecular biology of sunlight-induced carcinogenesis. DATA SOURCES: Journal and review articles, research studies, textbooks, and clinical practice. CONCLUSIONS: Basal cell and squamous cell carcinoma will occur in more than one million cases annually in the United States, and are highly curable when detected and treated early. During the last decade, significant progress has been made in elucidating the molecular basis of skin carcinogenesis and in identifying newer approaches for the management and treatment of these keratinocyte cancers. IMPLICATIONS FOR NURSING PRACTICE: Nurses can play crucial roles in decreasing the morbidity and mortality from the skin cancer epidemic by identifying and referring patients with lesions suspicious for basal cell and squamous cell carcinomas. PMID- 12638376 TI - Epidemiology of melanoma and nonmelanoma skin cancer. AB - OBJECTIVES: To describe the epidemiology of melanoma and nonmelanoma skin cancers. DATA SOURCES: Review and research articles, book chapters, and Surveillance, Epidemiology, and End Results (SEER) data. CONCLUSIONS: In 2002, an estimated 1.3 million Americans were diagnosed with skin cancer. Of these, 53,000 individuals were diagnosed with melanoma, the most common fatal form of skin cancer, and more than 7,000 Americans died of melanoma. Nonmelanoma skin cancer has the highest incidence of all cancers and the rise in the rate of cutaneous melanoma exceeds all other preventable cancers. IMPLICATIONS FOR NURSING PRACTICE: Nurses can act as case-finders and as advocates and educators for prevention of overexposure to ultraviolet radiation. Nurses should ascertain possible inherited risk and monitor patients for additional primary skin cancers. PMID- 12638378 TI - Diagnosis and management of stage I/II melanoma. AB - OBJECTIVES: To describe the clinical features and histologic subtypes of cutaneous melanoma; to review the diagnosis, clinical, and pathologic staging of melanoma and the associated prognostic factors; and to review the treatment and management of AJCC stage I and II melanoma. DATA SOURCES: Scientific and review articles, textbooks, and clinical practice. CONCLUSIONS: Management of melanoma depends on accurate diagnosis, staging, and interpretation of prognostic factors. The treatment of choice for stage I and II melanoma is surgery, ranging from simple excision to lymph node dissection. IMPLICATIONS FOR NURSING PRACTICE: Familiarity with the clinical features of melanoma assists nurses in the screening and early detection of melanoma. Knowledge of AJCC staging guides education regarding treatment and lifelong surveillance. PMID- 12638379 TI - Diagnosis and management of high-risk and metastatic melanoma. AB - OBJECTIVES: To identify treatment modalities and corresponding nursing implications for the high-risk and metastatic melanoma patient. DATA SOURCES: Texbooks, research articles, and professional experience. CONCLUSIONS: Recent advances in the field of melanoma include identification of prognostic factors, refinement of surgical techniques, and identification of effective adjuvant therapy. Novel therapies are currently under investigation. IMPLICATIONS FOR NURSING PRACTICE: Nurses play a vital role in the treatment of melanoma patients through education regarding their disease and treatment options, patient identification for clinical trials, and intensive monitoring and management for treatment-related side effects. PMID- 12638380 TI - Risk assessment and early detection of skin cancers. AB - OBJECTIVES: To describe a systematic method for skin cancer assessment, applying current standard practices for integration into nursing practice. To provide the fundamentals of performing a skin cancer assessment for the nonmelanoma skin cancers, basal cell carcinoma, and squamous cell carcinomas, and melanoma. Included in this discussion are risk profile calculations, mechanics of skin cancer assessment, descriptions of suspicious lesions, patient management, and follow-up. DATA SOURCES: Textbooks, research, review of the literature, and clinical experience. CONCLUSIONS: Skin cancer assessment is a skill that nurses can learn and implement into practice. IMPLICATIONS FOR NURSING PRACTICE: Knowledge and practice of good skin cancer assessment skills enhances nursing competence and positively influences patient outcomes. PMID- 12638381 TI - Skin cancer prevention: education and public health issues. AB - OBJECTIVES: To examine current public health initiatives related to skin cancer prevention and early detection, and public education issues. DATA SOURCES: Review articles, research reports, web sites, and government reports. CONCLUSIONS: Skin cancer remains a major public health problem. Efforts to decrease the morbidity and mortality associated with skin cancer are gradually shifting toward primary prevention. Primary prevention efforts should not only be limited to using sunscreen, but should also focus on reductions in ultraviolet light exposure. Secondary prevention efforts include professional skin examinations and skin self examination. Tertiary efforts focus on the prevention and detection of additional primary skin cancers. IMPLICATIONS FOR NURSING PRACTICE: Nurses can influence the public to practice primary, secondary, and tertiary prevention strategies for skin cancer. PMID- 12638383 TI - Psychological adjustment to the melanoma experience. AB - OBJECTIVES: To provide a review of the literature on the psychological adjustment to malignant melanoma. DATA SOURCES: Research reports and review articles. CONCLUSIONS: Three prominent themes evolved from the literature: (1) delineation of issues concerning long-term survivorship and quality of life following a diagnosis of melanoma; (2) coping with metastatic and end-stage melanoma; and (3) identification of the possible link between immunomodulation and the evolution of melanoma. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses must individualize their support and counsel according to the potential trajectory of the patient's melanoma experience. Nurses need to identify an individual's risk for psychological distress and help them enhance coping skills. PMID- 12638382 TI - Chemoprevention of skin cancer. AB - OBJECTIVES: To discuss the principles, science, and roles of nurses related to skin cancer chemoprevention. DATA SOURCES: Journal articles and federal government reports. CONCLUSIONS: The skin is a model organ for investigating cancer prevention processes that may be relevant to other organs as well. Agents that selectively target molecular hallmarks of skin carcinogenesis are under intense clinical and preclinical investigation. IMPLICATIONS FOR NURSING PRACTICE: Nurses play key roles in coordinating clinical trials, stimulating public awareness, and ensuring access to and acceptance of new agents. Nurses play a role in translational research by bridging the gap between technologic development and patient care. PMID- 12638384 TI - Informed consent and the scope of a physician's duty of disclosure. PMID- 12638385 TI - HIV transmission risks of females and males in paired partner relationships. AB - As the HIV/AIDS epidemic has evolved, heterosexual behaviors have come to be an increasingly important mode of transmission and rates of infection have increased faster among women generally and minority women in particular. Most of what we known about heterosexual risk taking is based on survey data collected from either women or men. These data have been useful for helping to understand different levels and types of risk taking by women and men. However, they provide little information about the levels of risk taking that exist for paired partners in main partner relationships, the types of risks each partner brings to the relationship and how the contribution of partner risks varies. Using data from the pretest of a pilot project designed to test a behavioral intervention to reduce HIV/AIDS transmission among main sex partners, the authors developed four subscales of sexual risk taking for each partner. The association between subscales within and between genders are examined and the subscales are combined to obtain measures of couple risks. Particular attention is given to the gender contribution of partners to couple risk and the differential contribution of types of risk to total risk. The data suggest that, even though men bring relatively higher risks to their main partner relationship than women, this varies inversely with the level of couple risk. PMID- 12638386 TI - Gaps between HIV/AIDS policies and treatment in correctional facilities. AB - In this article the authors examined correctional policy and its impact on the incidence of HIV/AIDS in prison population. Using data from the Florida Correctional System, they find that HIV/AIDS is still the leading cause of death. Improved treatment and care may have led to declines in AIDS-related mortality but the prison population continues to experience a much higher risk of mortality than he general population in spite of changes in the treatment and provision of care to infected patients. The dominance of HIV-related deaths indicates that treatment and voluntary testing policy have been ineffective. The authors argue that the persistence of HIV infections and AIDS-related deaths is largely attributable to continuing unequal distribution of health care resources between identified and unidentified HIV-infected inmates. Their analysis suggests that future changes in HIV/AIDS policy ib testing and treatment can contribute to improvement in health conditions of infected inmates. PMID- 12638387 TI - AIDS-related knowledge and behavior among married Kenyan men: a behavioral paradox? AB - The heterosexual character of HIV/AIDS transmission in sub-Saharan Africa, a context where men are dominant in sexual and reproductive matters, underscores the importance of assessing male behavior in sexual and related health arenas. Despite condom use being the recommended and expected behavioral response to knowledge about the fatal outcome of HIV/AIDS infection, use continues to be extremely low in sub-Saharan Africa. This article explores the relationship between various facets of knowledge about HIV/AIDS and condom use among married Kenyan men. The main finding is one of a significant interaction effect of the recognition that it is impossible to visually identify infected parties and one's perception of self-risk. Although neither is in itself significant, simultaneously recognizing that healthy-looking persons may be infected and perceiving that one is himself not at risk significantly reduces condom use among men. This finding--of an interaction effect--plausibly explains why a perception of self-risk, on its own, does not necessarily translate into safe behavior. After all, those who believe they can identify infected persons may think they are at low risk because they avoid contact with the infected and, in selecting partners they deem free of infection, they may be less inclined to use condoms. This finding has implications for how specific aspects of AIDS-related knowledge are imparted to communities and individuals as well as for our understanding of other health-related behaviors. PMID- 12638388 TI - Correlates of HIV/AIDS-related knowledge and preventive behavior of men in Africa. AB - Using the 1998/99 Ghana Demographic and Health Survey dataset, this article examines the behavior of men who think they are at risk for HIV/AIDS and those who indicate they have no AIDS risks. Using a theoretical framework that borrows from rationale choice theory and individual-level characteristics, the article explores the links between AIDS risks status and changes in preventive behavior. The results show high levels of AIDS-related knowledge among men in Ghana. In addition, the majority of the men indicated that they had changed their behavior in response to AIDS although they were more likely to cite the avoidance of multiple partners rather than the use of condoms in their sexual encounters. In the logistic regression models, education, marital status, and religion emerged as important predictors of changes in behavior. The fact that few men used condoms as an HIV/AIDS preventive method raises a number of policy issues, the implications of which are discussed in this article. PMID- 12638389 TI - Pharmacokinetic strategies in deciphering atypical drug absorption profiles. AB - Drug absorption is a very complex process that manifests itself through potential interaction with a host of physicochemical and physiological variables. Some factors that may affect the absorption processes include presystemic metabolism/efflux, the "absorption window" along the gastrointestinal tract, disease states, demographics (gender, age, ethnicity), and biopharmaceutical classification of solid dosage forms. Despite the complexity of the absorption processes, the analysis of the absorption kinetic data is mostly empirical, and the assumption of first-order absorption is axiomatic. Nevertheless, we often encounter irregular drug absorption profiles (such as double-peak, absorption window-type absorption profiles, etc.) that cannot be satisfactorily described by a simple first-order absorption process. The selection of an inappropriate absorption model would result in the misspecification of the pharmacokinetic model and subsequent erroneous prediction of the dosing regimen. This article presents several pharmacokinetic strategies in analyzing typical and atypical absorption profiles. The atypical absorption profiles discussed in this article include parallel first-order absorption, mixed zero-order and first-order absorption, Weibull-type absorption, absorption window with or without Michaelis Menton absorption, time-dependent absorption, and inverse Gaussian density absorption. In any event, intravenous drug concentration-time data are generally needed to avoid the ambiguousness in the absorption analyses. PMID- 12638391 TI - A model of pharmacology education: the experience of Istanbul Medical Faculty. AB - This article describes the pharmacology education program that has been applied since 1990 at the Istanbul Medical Faculty. In Turkey, medical education lasts 6 years after 11 years of general training. Each year, approximately 350 students join the Istanbul Medical Faculty. The education is mainly in conventional form: basic sciences and elementary clinical information are given mostly as didactic lectures accompanying practical courses for small groups, with each group consisting of about 50 students during the first 3 years. In the subsequent 2 years, students have several clinical clerkships, and the last year is an internship. Pharmacology, the bridge between basic and clinical sciences, has a special place in the medical training. Accordingly, the courses were expanded to 3, 4, and 5 years, the sum of all courses being approximately 140 hours. Pharmacology education took place along with basic sciences in the first years but with clinical sciences in the later years and is based on active learning methods. PMID- 12638390 TI - Reproducibility of dorsal hand vein responses to phenylephrine and prostaglandin F2 alpha using the dorsal hand vein compliance method. AB - Assessment of drug-induced venodilation by the dorsal hand vein compliance method requires stable constriction of the vein. This study was designed to investigate intra- and intersubject reproducibility of the venous preconstriction technique in response to phenylephrine and prostaglandin F2 alpha and to determine the influence of basal vein size. Twelve healthy male nonsmokers participated in a prospective cross-over study. Inter- and intrasubject variability was tested in response to phenylephrine and PGF2 alpha on different study days in the same hand vein. The dose of the respective constrictor causing approximately 80% constriction of the vein (ED80) was determined and infused for another 100 minutes. Actual vein size was measured every 5 minutes. Coefficient of variation and regression analyses were performed to analyze influence of vessel size on ED80 of the respective constrictor. Adjusted constriction levels were stable and well reproducible in all subjects. The intersubject coefficient of variation of ED80 ranged from 0.9% to 6.7% for phenylephrine and from 0.9% to 6.9% for PGF2 alpha. Whereas responses to phenylephrine were independent of basal vein diameter, there was a positive correlation between ED80 of PGF2 alpha and basal vein size. Thus, the hand vein compliance method is a suitable method to study dilatory responses in phenylephrine- or PGF2 alpha-constricted veins with considerable interindividual but small intraindividual variability. However, in such studies, phenylephrine appears to be a more reliable tool than PGF2 alpha. PMID- 12638392 TI - Pharmacokinetic/pharmacodynamic modeling of luteinizing hormone (LH) suppression and LH surge delay by cetrorelix after single and multiple doses in healthy premenopausal women. AB - Cetrorelix (CET) is a potent luteinizing hormone-releasing hormone (LH-RH) antagonist and is used to prevent premature ovulation in IVF (in vitro fertilization) procedures. The objective of the present study was to develop a pharmacokinetic/pharmacodynamic (PK/PD) model for the LH suppression and LH surge delay after single doses (SD) and multiple doses (MD) of CET in healthy premenopausal women without ovarian stimulation. CET was given by subcutaneous route (SD, 0.25, 0.5, or 1 mg) on cycle day 3 and as similar multiple once-a-day doses from cycle day 3 to day 16 in two consecutive menstrual cycles. The concentration-time data of CET and LH were used for PK/PD modeling. A two compartment model described the PK of CET with median terminal half-life estimates of 9.2 and 54.5 hours after SD and MD, respectively. An indirect response Emax model was used to describe the LH suppression and the LH surge delay. LH suppression was linked to plasma concentrations of CET, while the delay in the LH surge was linked to the PK of CET through a hypothetical effect compartment. Since the SD regimen on day 3 did not cause significant delay, these values were used as controls in the analysis of surge delay in MD data. The IC50 (for suppression) estimate was 0.73 ng/ml for SD, and EC50 (surge delay) was 1.42 ng/ml for MD. The PK/PD model adequately described the LH suppression and the surge delay. PMID- 12638393 TI - Pharmacokinetics of rosiglitazone in patients with varying degrees of renal insufficiency. AB - This study investigated the effect of varying degrees of renal insufficiency on the pharmacokinetics of rosiglitazone. Subjects were stratified by estimated creatinine clearance: normal (> 80 mL/min; n = 12), mild renal insufficiency (60 80 mL/min; n = 15), moderate renal insufficiency (30-59 mL/min; n = 18), and severe renal insufficiency not requiring dialysis (< or = 29 mL/min; n = 12). Plasma rosiglitazone concentrations and protein binding were determined after a single oral 8-mg dose of rosiglitazone. Total and unbound pharmacokinetic parameters were generated using noncompartmental methods. AUC, Cmax, and t1/2 data were analyzed separately by ANOVA to provide point estimates and corresponding 95% confidence intervals. The pharmacokinetics of rosiglitazone was not markedly affected by mild, moderate, or severe renal insufficiency. Slight increases (approximately 10%-20%) in mean unbound AUC0-infinity values were observed for each insufficiency group compared to the normal group but were not considered to be clinically relevant. Patients with severe insufficiency exhibited a 38% increase in mean fraction unbound, leading to an increase in total clearance, which resulted in a 19% to 24% lower mean total AUC0-infinity and Cmax values relative to the normal group. The rates of mild or moderate adverse events were similar for all groups; there were no severe adverse events. Impaired renal function does not markedly alter the pharmacokinetics of total or unbound rosiglitazone following a single dose of rosiglitazone. Therefore, the starting dose of rosiglitazone does not need to be adjusted in patients with renal impairment. Subsequent dose adjustments should be based on individual patient response. PMID- 12638394 TI - Effect of high-fat breakfast and moderate-fat evening meal on the pharmacokinetics of vardenafil, an oral phosphodiesterase-5 inhibitor for the treatment of erectile dysfunction. AB - The effects of food on the pharmacokinetics of vardenafil were examined in 25 healthy adult males. Single-dose vardenafil 20 mg was administered in a randomized four-way crossover design after an overnight fast (at 8 a.m.), after consumption of a high-fat breakfast (at 8 a.m.), on an empty stomach (at 6 p.m.), and after a typical moderate-fat evening meal (at 6 p.m.). Serial blood samples were analyzed for vardenafil and metabolite (M1) levels. When administered after an overnight fast and after a high-fat breakfast, vardenafil geometric mean Cmax was 17.14 and 14.0 micrograms/L, respectively, and AUC was 66.78 and 67.09 micrograms./h/L, respectively; the median tmax was 1 hour under fasting conditions and 2 hours with consumption of high-fat breakfast. When administered in the evening on an empty stomach and after a moderate-fat meal, vardenafil geometric mean Cmax was 14.22 and 13.04 micrograms/L, respectively, and AUC was 51.97 and 59.12 micrograms.h/L, respectively. The median tmax was 1 hour after fasting or a moderate-fat meal in the evening. All treatments were well tolerated. Thus, while a high-fat meal may alter Cmax slightly and delay the absorption up to 1 hour, a moderate-fat meal has no clinically relevant effect on vardenafil pharmacokinetics. Dosage changes are not warranted based on the wide therapeutic index and the efficacy observed with vardenafil in Phase III studies that were not restricted with respect to food. PMID- 12638395 TI - Single- and multiple-dose pharmacokinetics of etoricoxib, a selective inhibitor of cyclooxygenase-2, in man. AB - The single- and multiple-dose pharmacokinetics of etoricoxib, a selective inhibitor of cyclooxygenase-2, were examined in two clinical studies. Single-dose pharmacokinetics--including dose proportionality, absolute bioavailability of the highest dose-strength (120-mg) tablet, and the effect of a high-fat meal on the bioavailability of that tablet--were investigated in a two-part, open, balanced crossover study in two panels of healthy subjects (12 per panel). Steady-state pharmacokinetics were investigated in an open-label study in which 24 healthy subjects were administered 120-mg single and multiple (once daily for 10 days) oral doses of etoricoxib tablets. The pharmacokinetics of etoricoxib were found to be consistent with linearity through doses at least twofold greater than the highest anticipated clinical dose of 120 mg. Etoricoxib administered as a tablet was rapidly and completely absorbed and available; the absolute bioavailability was estimated to be 100%. A high-fat meal decreased the rate of absorption without affecting the extent of absorption of etoricoxib; therefore, etoricoxib can be dosed irrespective of food. Steady-state pharmacokinetics of etoricoxib, achieved following 7 days of once-daily dosing, were found to be reasonably predicted from single doses. The accumulation ratio averaged 2.1, and the corresponding accumulation t1/2 averaged 22 hours, supporting once-daily dosing. Etoricoxib was generally well tolerated. PMID- 12638396 TI - Pharmacokinetics of pregabalin in subjects with various degrees of renal function. AB - The objectives of this study were to determine the single-dose pharmacokinetics of pregabalin in subjects with various degrees of renal function, determine the relationship between pregabalin clearance and estimated creatinine clearance (CLcr), and measure the effect of hemodialysis on plasma levels of pregabalin. Results form the basis of recommended pregabalin dosing regimens in patients with decreased renal function. Thirty-eight subjects were enrolled to ensure a wide range of renal function (CLcr < 30 mL/min, n = 8; 30-50, n = 5; 50-80, n = 7; and > 80, n = 6). Also enrolled were 12 subjects with renal impairment requiring hemodialysis. Each subject received 50 mg of pregabalin as two 25-mg capsules in this open-label, parallel-group study. Pregabalin concentrations were measured using previously validated liquid chromatographic methods. Pregabalin pharmacokinetic parameters were evaluated by established noncompartmental methods. Pregabalin was rapidly absorbed in all subjects. Total and renal pregabalin clearance were proportional (56% and 58%, respectively) to CLcr. As a result, area under the plasma concentration-time profile (AUC) and terminal elimination half-life (t1/2) values increased with decreasing renal function. Pregabalin dosage adjustment should be considered for patients with CLcr < 60 mL/min. A 50% reduction in pregabalin daily dose is recommended for patients with CLcr between 30 and 60 mL/min compared to those with CLcr > 60 mL/min. Daily doses should be further reduced by approximately 50% for each additional 50% decrease in CLcr. Pregabalin was highly cleared by hemodialysis. Supplemental pregabalin doses may be required for patients on chronic hemodialysis treatment after each hemodialysis treatment to maintain steady-state plasma pregabalin concentrations within desired ranges. PMID- 12638397 TI - Multiple-dose escalation, safety, and tolerability study of wood creosote, the principal active ingredient of seirogan, an herbal antidiarrheal medication, in healthy subjects. AB - Seirogan, an herbal medicine containing wood creosote (CAS 8021-39-4), a mixture of simple phenolic compounds, has been marketed for the past century in Asia for the treatment of acute diarrhea and associated symptoms, such as abdominal discomfort and cramping. The present study was designed to assess the safety and tolerability of an anticipated acute antidiarrheal dosing regimen. Sixty healthy males were randomized into five groups of 12 subjects each (9 wood creosote; 3 placebo) to receive 45-, 90-, 135-, 180-, and 225-mg tablets every 2 hours for five doses. Serial sitting and standing vital signs, ECG rhythm strips, and continuous telemetry monitoring were obtained predose and for 24 hours after the first dose. Clinical laboratory tests and 12-lead resting ECGs were obtained predose and 24 hours postdose. Of the subjects, 27% (12/45) receiving wood creosote and 27% (4/15) receiving placebo reported adverse events. The most common adverse events were altered taste and somnolence, reported more often with 180- and 225-mg doses. Wood creosote had no clinically significant effects on vital signs, ECG intervals or interpretations, or clinical laboratory tests. No clinically significant or serious dysrhythmias were reported on continuous telemetry monitoring. It was concluded that oral doses of wood creosote 45 to 225 mg every 2 hours for up to five doses were safe and well tolerated in 45 healthy subjects. Wood creosote doses ranging from 45 to 135 mg per dose, which are commonly administered antidiarrheal doses in Asia, were associated with minimal side effects. PMID- 12638398 TI - Topical hemostatic effect of a common ornamental plant, the geraniaceae Pelargonium zonale. AB - Geranium has been traditionally used as a local hemostatic medicine in some Andean regions, but this effect has not been tested in controlled experiments. In the present report, the leaves of a geraniaceae (Pelargonium zonale) were tested on a bleeding rat model. The bleeding time was 50% shorter in the geranium leaf juice treatment group (18.10 +/- 2.03 min) and 80% shorter in the geranium crushed-leaf group (7.10 +/- 0.88 min) than in the control (nontreatment) group (37.6 +/- 3.04 min), p < 0.0001. Bleeding time with guava (Psidium guajava) crushed leaves (39.90 +/- 1.54 min) was not different from the control group. A proved hemostatic agent, gelatin sponge, had a similar effect as geranium juice (16.7 +/- 3.32 min) in the same animal model. A buffer solution at pH 3 (the same pH as the geranium leaf extract) did not have any hemostatic effect, and the bleeding time (39.3 +/- 2.71 min) was not different from the control group. The dilution 1:4 geranium leaf juice at pH 3(25.6 +/- 3.08 min) or pH 5 (28.8 +/- 3.98 min) still had a statistically significant hemostatic effect. The results confirm the hemostatic effect of P. zonale leaves and show that it is similar (geranium leaf juice) or better (crushed geranium leaves) than the hemostatic effect of a commercial hemostatic sponge. It seems that the hemostasis caused by P. zonale extract leaves is not due to its low pH. The potential benefits as a new, inexpensive, safe, and easily available natural topical hemostatic agent are discussed. PMID- 12638399 TI - Randomized, double-blind, placebo-controlled comparison of the analgesic efficacy of oxycodone 10 mg/acetaminophen 325 mg versus controlled-release oxycodone 20 mg in postsurgical pain. AB - This randomized, controlled trial compared the analgesic efficacy and safety of the new oxycodone 10-mg/acetaminophen 325-mg formulation (Percocet) for the treatment of acute pain following oral surgery with double the dose of oxycodone alone (controlled-release [CR] oxycodone 20 mg [OxyContin]). A total of 150 male and female patients with > or = 2 full or partial bone-impacted mandibular molars, at least moderate persistent pain, and moderate trauma received a single dose of combination agent, CR oxycodone, or placebo following oral surgery and rated pain intensity and pain relief over the next 6 hours. The intent-to-treat population comprised 141 patients (55 on combination agent, 56 on oxycodone, and 30 on placebo). Combination agent and CR oxycodone were significantly superior to placebo for all efficacy measures. Combination agent was statistically superior to CR oxycodone in four of five outcome measures of pain intensity and pain relief (PPID, PPAR, SPID, and SPRID). It also provided a faster onset and 24% reduction in the number of patients reporting treatment-related adverse events compared with twice the dose of opioid alone. This new formulation offers the combination of two analgesic drugs with complementary mechanisms of action, which results in enhanced analgesia, an "opioid-sparing" effect, and an improved side effect and safety profile. PMID- 12638401 TI - "New new" things for revenue-cycle improvement. PMID- 12638400 TI - Ketoprofen 75 mg qid versus acetaminophen 1000 mg qid for 3 days on swelling, pain, and other postoperative events after third-molar surgery. AB - A randomized, double-blind, within-patient, crossover study was carried out after bilateral "identical" surgical procedures using local anesthesia only. A 3-day tablet regime of racemic ketoprofen 75 mg or acetaminophen 1000 mg qid (x 4) was given starting 3 hours after surgery. Ketoprofen reduced objectively measured swelling 27.8% (p < 0.04) better than acetaminophen 3 days after surgery and 70.8% (p < 0.02) better than acetaminophen 6 days after surgery. The pain intensity (PI) was lower after ketoprofen than after acetaminophen from 2 to 6 hours after the first drug intake (all p-values < or = 0.03). Sum PI during the first (SUMPI3.5-6, p = 0.003) and second dose intervals (SUMPI6.5-9, p = 0.007) was lower for ketoprofen than for acetaminophen but not different for the third dose interval (SUMPI9.5-11, p = 0.53). Ketoprofen was a more effective analgesic than acetaminophen on the day of surgery (SUMPI3.5-11, p = 0.005). There was no difference (p > 0.05) between the treatments with respect to mouth opening, drug preference, global evaluation, or adverse reports. Adverse reports included stomach pain and diarrhea in both treatment groups. Ketoprofen 75 mg x 4 for 3 days reduces subjectively assessed pain and objectively measured swelling (i.e., anti-inflammatory effect) following third-molar surgery. PMID- 12638403 TI - [Pulmonary diseases in patients with peritonitis]. PMID- 12638402 TI - [Role of organic nitrates in therapy of coronary heart disease in terms of clinical cost-effectiveness]. PMID- 12638404 TI - [Pulmonary tuberculosis in diabetic patients]. PMID- 12638405 TI - [Obstructive sleep apnea: clinical significance and correlations with arterial hypertension]. AB - Obstructive sleep apnea (OSA) occurs rather frequently. It often combines with arterial hypertension (AH) and contributes to development and course of such severe conditions as stroke, myocardial infarction, arrhythmia, sudden death in sleep. Lack of adequate knowledge of relevant symptoms, cause-effect relationships leads to mistakes in management of patients. AH patients with OSA should receive combined treatment including hypotensive drugs and correctors of sleep respiratory disorders. PMID- 12638406 TI - [Non-pharmacological correction of arterial pressure in hypertensive patients with a pacemaker]. AB - Frequency-dependent hypotensive effect of cardiac pacing (CP) regarding type of circulation and stimulation regime under programmed changes of impulse frequency from 50 to 90 imp/min was studied in 92 patients (53 males, 39 females) with essential hypertension of the first, second and third degree and isolated systolic arterial hypertension. The rise of the above frequency over 80 imp/min was accompanied with a significant fall in systolic arterial pressure (SAP) in patients with hyperkinetic circulation while heart rhythm under 60 imp/min resulted in a significant lowering of diastolic arterial pressure (DAP) in patients with eukinetic and hypokinetic type of circulation. In regime VVI both SAP and DAP lowering was more pronounced than in AAI regime. The most prominent hypotensive effect was seen in VVI-stimulation complicated by ventriculoatrial conduction. Reprogramming of impulse frequency normalized arterial pressure in 43.3% patients with hyperkinetic type of circulation. Thus, a frequency-dependent hypotensive effect of programmed CP can be used for non-pharmacological correction of arterial hypertension and optimization of effectiveness of anti hypertensive treatment in paced patients. PMID- 12638407 TI - [Echocardiographic evidence on development of chronic cor pulmonale in asthmatics]. AB - Early detection of chronic cor pulmonale (CCP) is essential for diagnosis of the patient's condition, choice of adequate therapy, improvement of the prognosis. The study of parameters of pulmonary hemodynamics, right and left heart in 125 patients with bronchial asthma determined how CCP is formed in such patients. PMID- 12638408 TI - [Vasopressin and renin-aldosterone system in patients with arterial hypertension]. AB - Levels of vasopressin, aldosterone and plasma renin activity were studied in patients with arterial hypertension. It is shown that progression of the disease dysregulates neurohormonal systems responsible for arterial pressure control. Activation of vasoconstrictive systems of vasopressin and aldosterone synthesis involved in retention of fluid in the body goes in parallel to lowering of renin secretion. The above changes may be considered as a compensatory mechanism of arterial hypertension stabilization. PMID- 12638409 TI - [Cardio-vascular system condition in respiratory mycoplasmosis]. AB - The study enrolled 53 patients (29 males, 24 females) with respiratory mycoplasmosis aged 15-88 years. Most of them were 59 years of age and younger. In 1/3 of the patients the diseases started with symptoms of acute respiratory viral infection, in 2/3 of them--with pulmonary affection. Pneumonia was diagnosed in 50 patients (94.3%), acute bronchitis--in 3 patients. ECG changes were registered in about half of the examinees who had no cardiac complaints. 25 of them had alterations in the end part of the ventricular ECG complex; rhythm and conduction disturbances occurred rarely. Mycoplasmosis patients suffering from ischemic heart disease (IHD) had stable ECG changes while in those free of IHD the changes were short. Myocardial necrosis foci were absent. Cardiac damage comparison in patients with respiratory mycoplasmosis and in other acute respiratory infections has found that cardiovascular system suffers less in acute mycoplasmosis. These data are useful in differential diagnosis of myocardial infections. PMID- 12638410 TI - [Clinical presentation of connective tissue dysplasia in adults]. AB - Connective tissue dysplasia (CTD) markers (anthropometric parameters, joint hypermobility, state of the skin, osteomuscular system, etc.) as well as external respiration function, echocardiography, occlusive plethysmography data were assessed in 402 healthy subjects (142 males and 260 females, mean age 28.4 +/- 0.5 years). Four basic CTD symptom complexes were identified: ligamentous articular, cutaneous, osteomuscular and cardial. Control group consisted of subjects free of CTD markers (n = 61). Combination of the above four symptom complexes occurred in 8.5% examinees. These show psychoemotional disorders, impaired function of external respiration, decreased size of the heart, low arterial pressure, affection of capillary and venous circulation. PMID- 12638411 TI - [Changes in bile lithogenicity and metabolism of connective tissue in chronic cholecystitis]. PMID- 12638412 TI - [Function of the liver and gallbladder after subtotal gastric resection and gastrectomy with the creation of an artificial stomach]. AB - A comparative ultrasound and radiological investigation of gallbladder and hepatic tree function was made in patients after operation with creation of artificial stomach, Billroth I and Roux operations. The control group consisted of 30 unoperated patients. Scintigraphy discovered gallbladder dyskinesia in 12 (18.7%) patients of the study group and in 6 (20%) controls. Ultrasonography--in 15 (33.3%) patients of the study group, 5 (25%) patients after distal resection of the stomach with pyloriform gastroduodenoanastomosis, 18 (51.47%) patients operated by Roux and 5 (16.6%) controls, respectively. It is shown that normal transduodenal passage of food has a positive effect on gallbladder and liver functions. PMID- 12638414 TI - [Use of amixin in the therapy and prevention of some viral infections]. AB - The trial of oral interferon inductor amixin for effectiveness in chronic viral hepatitis (CVH) and hemorrhagic fever with renal syndrome (HFRS) has shown that amixin in CVH improved general condition of the patients, removed yellowness of the skin and sclera, normalized activity of aminotransferases and blood bilirubin level. Virus replication was stopped in 25 and 1.6% in CVHB and CVGC, respectively. Amixin in HFRS was effective if received early. Preventive amixin therapy in population groups with high HFRS risk prevents development of HFRS and acute respiratory viral infection. PMID- 12638413 TI - [Renal hemodynamics in patients with systemic lupus erythematosus as shown by renal scintigraphy]. AB - Dynamic scintigraphy (DS) provides qualitative and quantitative assessment of renal circulation in patients with different diseases. Few data are still available on application of DS of the kidneys in patients with systemic lupus erythematosus (SLE). Renal hemodynamics studies with DS are not described in SLE patients with disturbances of renal metabolism and/or antiphospholipid syndrome (APS). We examined renal hemodynamics in SLE patients with renal disease, arterial hypertension, impaired lipid metabolism, APS. We examined 65 patients with confirmed diagnosis of SLE with 99mTc DS and estimated effective renal blood flow. The latter was damaged more in SLE patients with symptoms of renal disorders. Arterial hypertension deteriorated renal hemodynamics. In hypertensive patients with affected kidneys angionephroscintigraphic parameters changed most significantly. Renal hemodynamics in disturbed lipid metabolism was worse than in normal one but the difference was not significant. Angionephroscintigraphy findings were worse in patients with APS than in those with renal lesion and arterial hypertension but free of APS. PMID- 12638415 TI - [Experience of COB-school for patients with chronic obstructive bronchitis]. AB - The results of training of chronic obstructive bronchitis (COB) patients according to the educational program "COB-school" are presented. The program was developed basing on the results of examination of 132 COB patients (80 males and 52 females) which included assessment of clinical picture, external respiration function, histological verification, standard multifactorial personality investigation, Luscher's test, specially designed questionnaire to diagnose social status. The findings demonstrate that COB patients respond to their disease with inadequate emotions, their psychological reactions are destructive, they suffer from social maladaptation. It is also shown that COB belong to diseases with abnormal pulmonary ventilation which is stressogenic and, consequently, treatment discipline of such patients is low. Methodology of the educational program rests on psychological laws of coping with stressogenic situation. The informative blocks of the program were constructed basing on these laws while the analysis of personality disorders provided grounds for optimal information presentation. The program covered 64 female and 50 male learners with COB. The effect of the program was evaluated by several clinical indices and reached 66.8 and 66.0% in females and males, respectively and a total of 66.4%. Thus, educational program with psychological and social correction noticeably improves discipline of COB patients' treatment. PMID- 12638416 TI - [Differential diagnosis of nonspecific aortic arteritis]. AB - The article presents diagnostic criteria for nonspecific aortic arteritis proposed by K. Ishikawa (1988), American Rheumatology Society (1990), algorithm of differential diagnosis with giant cell arteritis, obliterating thrombotic angiitis, Marfan's syndrome, atherosclerosis of major vessels, etc. PMID- 12638417 TI - [Painless myocardial ischemia as a risk factor of constantly relapsing paroxysms of cardiac fibrillation]. AB - A case is reported illustrating relationship between marked coronary atherosclerosis with development of painless myocardial ischemia (PMI) and constantly recurrent paroxysms of cardiac fibrillation (CF). A male 62-year-old patient suffering from paroxysmal CF received cordaron in a dose 200 mg twice a day. The treatment was ineffective until a comprehensive examination (thyroid hormones, echocardiography, Holter ECG monitoring, treadmill test, CT of the coronary arteries) found PMI episodes and severe stenosing atherosclerosis and the treatment was changed for sotalex (80 mg twice a day). Sotalex treatment was effective and stopped recurrences of CF. PMID- 12638418 TI - [Effect of beta-interleukin on the content and proliferation of hemopoietic precursor cells in intact and irradiated long-term cultures of the bone marrow]. AB - Effects of beta-leukine (human recombinant interleukin-1 beta) on the number and proliferative activity of early hemopoietic precursor cells (CFU-GM) and colonystimulating activity (GM-CSF) were studied in long-term cultures of the bone marrow. Studied were also recovery of CFU-GM, total number of myelocaryocytes and content of immature and mature granulocytes in response to beta-leukine after gamma-radiation of cultures in a dose 2.0 Gy. It was found that beta-leukine enhanced GM-CSF induction and elevated content of early CFU-GM in non-radiated long-term cultures of the bone marrow maximum 20 hours after introduction. 20 hours after introduction of beta-leukine proliferative activity of CFU-GM reached its maximum. The study of postradiation recovery in administration of beta-leukine in a dose 0.01 mcg/ml 10 min after radiation of long-term cultures of the bone marrow registered no significant stimulating effect of the drug on CFU-GN recovery, recovery of the number of myelocaryocytes and content of young immature and mature granulocytes. PMID- 12638420 TI - [Functions of peritoneal macrophages during adaptation to dosed heat factor]. AB - Experiments on mice were made to study functional activity of peritoneal macrophages in adaptation of the animals to dosed heat factor by a phagocytic ability of the cells, chemiluminescence (CL), activity of acid phosphatase and phagocyte production of lymphocyte-activating factors (LAF). Daily overheating of mice (20 min, 43-44 degrees C) for 5, 10, 20 and 30 days resulted in suppression of macrophage function. The deepest depression of phagocyte functional activity was observed on overheating day 10-20. This was evident from subnormal phagocytic ability of the cells, suppression of CL response and activity of acid macrophage phosphatase, low production of LAF. Thermotraining led to recovery of macrophage function as the studied parameters returned to control 40 days after overheating onset. PMID- 12638421 TI - [Blood oxygen transport and pro-oxidant-antioxidant state during liver reperfusion]. AB - Indices of blood oxygen transport (hemoglobin-oxygen affinity, pCO2, pH, pO2, etc.) and prooxidant-antioxidant state (Schiff bases, conjugated dienes, catalase, retinal, alpha-tocopherol) were measured in rabbit blood and the liver during postischemic reperfusion. Hepatic ischemia was induced for 30 min by ligation of a hepatica propria, and reperfusion lasted for 120 min. Hepatic ischemia worsened blood oxygen transport. Restoration of arterial blood flow did not result in improvement of oxygen delivery. Moreover, marked metabolic acidosis was observed throughout 2 hr of reperfusion. Ischemia induced a shift of oxyhemoglobin dissociation curve to the right. This shift persisted after restoration of hepatic arterial blood flow facilitating increased oxygen transport to tissues. Changes in blood oxygen transport during hepatic ischemia/reperfusion were accompanied with high activity of free radical processes. During reperfusion, the largest increase in content of lipid peroxidation products and the greatest fall of some antioxidant levels except catalase were observed indicating impairment of liver prooxidant-antioxidant balance. The results showed that activation of lipid peroxidation and a decrease in some antioxidant levels during hepatic reperfusion were associated with lowering of hemoglobin-oxygen affinity and suggest participation of the latter in impairment of prooxidant-antioxidant balance. PMID- 12638419 TI - [Effect of adrenalectomy in pregnant mice on activity and spectrum of alkaline phosphomonoesterase in placenta and fetal organs]. AB - Unilateral adrenalectomy in mice on pregnancy day 14 stimulates activity of alkaline phosphomonoesterase in placenta on pregnancy day 21. This is often accompanied with changes in the enzyme activity in fetal liver, kidneys, bone, intestine. Additional isoforms of alkaline phosphomonoesterase appear in the electrophoretic spectrum. PMID- 12638422 TI - [Ammonia-neutralizing function of hepatocytes after experimental liver resection]. AB - The experimental study has established that a 15-20% resection of a hepatic mass in healthy rats inhibits glutamine formation in hepatocytes and stimulates its decomposition in liver cells promoting ammonia accumulation in the operated liver. The hepatocyte urea synthetizing cycle can not prevent accumulation of ammonia in the liver and raise their urea content even if arginase activity is high. Inhibition of GDG activity in the operated liver not only limits formation of ammonia during glutamate deamination but also results in glutamine deamination inhibition in hepatocytes. These disorders of hepatocytic ammonia neutralizing function persist for 21 days after hepatic resection. PMID- 12638424 TI - [Metabolism of sialoglycoproteins of the stomach of rats with experimental diabetes]. AB - Experiments on white non-inbred male rats studied gastric sialoglycoprotein metabolism. It was found that rats with experimental (alloxan) diabetes exhibit enhanced catabolic processes in sialoglycoprotein metabolism characterized by elevated concentrations of free, oligobound sialic acids and sialidase activity in insignificantly elevated concentrations of protein-bound sialic acids. PMID- 12638425 TI - [Space-time organization of bone tissue remodeling in vertebral bodies after femur osteotomy]. AB - Roentgenometric and roentgenodensitometric chronobiological characteristics of space and time remodeling of the bone tissue in tail vertebra XI and XII after osteotomy of the right femur are described. Examination of 179 white non-inbred male rats revealed circadian periodicity of vertebral bone tissue remodeling. Dynamics of asymmetry of this process also has a circadian periodicity. PMID- 12638423 TI - [Effect of 1,2,4-trimethylbenzene inhalation on energy metabolism in experimental animals]. AB - The authors' investigations show that red cells and tissues of animals exposed to a single 4-h inhalation of vapors of 1,2,4-trimethylbenzene in concentration 10 mg/m3 contained reduced amount of ATP, accumulated amounts of ADP, AMP in active inclusion of 32-P in the structure of these compounds. Activation of ATPase is accompanied with a rise of Na and P concentrations and fall in K, Mg and Ca ions concentrations. In blood plasm there were lower levels of Na and Ca and higher levels of K, Mg and P ions. In exposure to ecotoxicant in concentration 0.01-0.1 and 1.0 mg/m3 the changes were insignificant. PMID- 12638426 TI - [Blood and oxygen supply of visual cortex analyzer in children with normal vision and myopia]. AB - Blood and oxygen supply, respiration and cerebral cortex function especially those cortical parts in which visual analyzer is represented were studied in children and adolescents with myopia and accommodation spasm. It was found that respiration function and oxygen regime in myopia children and adolescents are 2-3 years behind those in normal children and adolescents. Electroencephalogram data show that index of alpha-rhythm in the occipital lobes (especially in the left lobe) and amplitude of alpha-rhythm in the right lobe are lower than in healthy subjects. Delta waves prevail on EEG of myopia patients, especially in the front lobes. Indirect signs of general cerebral dyscirculatory symptoms and dysfunction of the subcortical structures were registered as well as moderate changes in bioelectric activity of the cerebral cortex. 20-day interval hypoxic training in children and adolescents raised efficacy of the external respiration and circulation, normalized bioelectric activity. The majority of the patients improved their visual acuity up to complete normalization. PMID- 12638427 TI - [Clinico-morphological correlations and pathophysiological mechanisms in acute appendicitis]. PMID- 12638428 TI - [Treatment of dyslipoproteinemia by systemic enzyme therapy in experimental glomerulonephritis]. AB - Patients with chronic glomerulonephritis (CG) develop disturbances of lipid blood spectrum leading to additional damage to renal structure. The existent methods of pathogenetic therapy have no effect on lipid imbalance. Recently, many autoimmune diseases have been treated with systemic enzyme therapy (SET). The authors studied SET effect in disturbed lipid metabolism in experimental glomerulonephritis. Experimental animals showed morphological and biochemical changes similar to those in CG of man. SET reduced renal tissue damage and symptoms of dyslipoproteinemia. PMID- 12638429 TI - [Academician Aleksandr Aleksandrovich Bogomolets]. PMID- 12638430 TI - [Patient with bradycardia. When will he need a pacemaker?]. PMID- 12638431 TI - [Established blood pressure drug wins out in a comparative study. In hypertension, a diuretic first!]. PMID- 12638432 TI - [Consensus recommendations for influenza therapy. Proper administration of new influenza drugs]. PMID- 12638433 TI - [New laser equipment to improve diagnosis and follow-up. What can the new rheumatoid scanner do? (interview by Dr. Thomas Meissner)]. PMID- 12638434 TI - [The physician in political health care reform discussion. Scapegoat and target of abuse]. PMID- 12638435 TI - [Only 1 injection per week brings a 60% chance of healing. New therapy recommendations in chronic hepatitis C]. AB - Antiviral therapy can now eliminate the hepatitis C virus in some 60% of all hepatitis patients. The results of recent studies have led to new recommendations for treatment: patients with HCV genotype I should be treated with pegylated interferons and 1000 or 1200 (patients weighing > 75 kg) mg ribavirin for 48 weeks, while patients with genotypes 2/3 should receive only 800 mg ribvavirin (irrespective of body weight) for 24 weeks. Data provided by two large drug approval studies with PEG interferons alpha 2a and 2b show that the initial decrease in HCV-RNA determines the sustained viral response. Patients with genotype 1 have only a minimal chance (0-3%) of a sustained viral response if HCV RNA is not reduced by at least a factor of 100 by the 12th week of treatment, and in these patients, treatment should then be discontinued. Pre-treatment determination of HCV genotype and HCV-RNA is mandatory to establish the chances of attaining a sustained viral response. Liver biopsy is necessary to determine the stage of disease and the urgency of treatment. In young persons with biochemical and histological inflammatory activity treatment is usually indicated. In asymptomatic older patients with no liver fibrosis, however, in particular those with a limited chance of a sustained viral response or concomitant disease, the indication for antiviral therapy is doubtful. PMID- 12638436 TI - [Managing hepatitis C patients. Much work, little compensation]. PMID- 12638437 TI - [Follow-up in kidney transplantation. Which complications are likely?]. AB - Kidney transplantations are being performed in every increasing numbers, and, in addition to specialist centres, nephrologists, general practitioners and urologists are more frequently being involved in the provision of aftercare. The major urological complications seen after transplantation include ureteral stenosis and necrosis, incontinence and impotence, lymphoceles, arterial stenosis, infections of the urinary tract and adnexa, as well as--the most serious complication--malignant tumours. In addition, the immunosuppression required by these patients is often associated with secondary disorders such as hypertension, steroid-related diabetes, hyperlipidemia and infections. Against the background of progressive organic deficiency it is of particular importance to identify such complications and to treat them adequately, or to prevent them from occurring, thus ensuring the longest possible survival of transplant and patient and improved quality of life. PMID- 12638438 TI - [The acute phase of stroke. Treating hypertension or not?]. PMID- 12638439 TI - [Large round lesion in the liver. Malignant or benign? Cavernous hemangioma]. PMID- 12638440 TI - [Losartan in isolated systolic hypertension--convincing results of a LIFE sub group study]. PMID- 12638441 TI - [Even in advanced heart failure, hawthorn improves physical capacity]. PMID- 12638442 TI - [AT1 blocker therapy of hypertension in seniors. Added protection for cerebral blood vessels?]. PMID- 12638443 TI - [Rapid, strong and long-term effectiveness. Coxib also effective in toothache]. PMID- 12638444 TI - [Early, but not unnecessarily frequent administration. What is proper timing for triptan drugs?]. PMID- 12638445 TI - [Deficits in preventing recurrence in alcoholic patients. Withdrawal--and immediately back to the bottle]. PMID- 12638446 TI - [Highly effective and tolerable protease inhibitor. New trump card against HIV]. PMID- 12638447 TI - [In acute pain, self-medication with ASS is reliable]. PMID- 12638448 TI - [Treatment with NMDA antagonist controls neuron death. Even in severe dementia it is not too late]. PMID- 12638449 TI - [Moxonidine is advantageous for hypertensive type 2 diabetic patients. Therapy of hypertension also lowers HbA1c]. PMID- 12638450 TI - [Fewer repeat measurements, reduced costs. Reliable blood glucose test strips]. PMID- 12638452 TI - Leptospirosis. PMID- 12638451 TI - XXth SNA Biennial Conference, Ernakulam in Kerala from November 26-30, 2002. PMID- 12638453 TI - Development of flow sheet for the patients with eclampsia (admitted in eclampsia unit of a selected hospital). PMID- 12638454 TI - Conference on global nursing partnership. 'Strategies for a sustainable nursing workshop'. PMID- 12638455 TI - A clinician's guide to surgical fires. How they occur, how to prevent them, how to put them out. AB - Surgical fires--fires that occur on or in a surgical patient--are a rare but devastating complication of surgical procedures. They can happen during almost any kind of operation and can have dire consequences for both the patient and the hospital. The risk of surgical fires can be minimized as long as everyone on the surgical team understands how fires start and what to do when they occur. It's important to recognize the interaction among ignition sources, oxidizers, and fuels--the three sides of the classic fire triangle. What's more, staff members need to be alert to "hidden" fire risks, such as unsuspected oxygen-enriched atmospheres and less-than-obvious fuels. They also need to be drilled in the correct steps for responding to fires. In this guide, we describe the elements of surgical fires, including the types of equipment and materials that make up each side of the fire triangle. Through a series of case histories, we illustrate the many ways in which fires can start in the operating room. We also provide a detailed list of prevention methods. Finally, we describe the steps to take and the equipment to use--and not use--when a fire does occur. PMID- 12638457 TI - Medical gas fires: does your staff know how to recognize and extinguish them. PMID- 12638456 TI - Educational videos on surgical fires. AB - ECRI has been investigating, reporting on, and teaching about surgical fires for about 30 years. Our experience has shown that most of these fires could have been avoided had the staff been adequately educated about the hazards. Educational videos are just one tool that hospitals can use to provide the fire-safety training that surgical staff require. In this Evaluation, we look at eight videos covering fire safety in the operating room (OR). Our examination focuses on how well these videos address surgical fires in particular, since these can be the most devastating type of OR fire. We rate each video on how effectively it meets the fire-safety training needs of today's healthcare workers. For an educational fire safety video to earn at least an Acceptable rating, it must clearly and correctly explain what surgical fires are, how they occur, how they can be avoided, and how staff should respond if a fire occurs. Only three of the videos meet those criteria. The rest either contain errors, are missing information, or make hazardous recommendations and are rated Not Recommended or Unacceptable. PMID- 12638458 TI - Using carrying cases to store endoscopes can lead to patient infection. PMID- 12638459 TI - Inadequate wiring on Fresenius 2008H hemodialysis units. PMID- 12638460 TI - Piped nitric oxide delivery systems. PMID- 12638461 TI - Soy isoflavones: a safety review. AB - Soy isoflavones have been a component of the diet of certain populations for centuries. The consumption of soy generally has been considered beneficial, with a potentially protective effect against a number of chronic diseases; because of their estrogenic activity, however, negative effects of isoflavones have been postulated. This review examines the literature associated with the safety of soy isoflavones, including dietary soy isoflavone exposure data of populations with high soy intakes, human studies in which soy protein or isoflavones were provided, and toxicologic studies investigating the potential genotoxicity, carcinogenicity, and reproductive and developmental toxicity of soy isoflavones. Whereas results in some studies are limited or conflicting, when viewed in its entirety, the current literature supports the safety of isoflavones as typically consumed in diets based on soy or containing soy products. PMID- 12638462 TI - Not all injury-induced muscle proteolysis is due to increased activity of the ubiquitin/proteasome system: evidence for up-regulation of macrophage-associated lysosomal proteolysis in a model of local trauma. AB - A characteristic response to injury is a dramatic loss of skeletal muscle protein owing to increased muscle protein breakdown. Over the past decade, numerous studies have indicated that up-regulation of the ubiquitin-proteasome system is a common mechanism underlying such injury-induced muscle proteolysis. However, a recent study using a single-impact trauma to the gastrocnemius muscle found that, although the rate of muscle proteolysis was dramatically increased, the ubiquitin proteasome system was not involved. Rather, an increase in lysosomal activity, through infiltration of the damaged tissue by mononuclear macrophages, is responsible for the high rates of protein breakdown. PMID- 12638463 TI - Current directions in hemochromatosis research: towards an understanding of the role of iron overload and the HFE gene mutations in the development of clinical disease. AB - Since the discovery of a candidate gene (HFE) thought to be involved in the development of hereditary hemochromatosis, there has been much interest in the potential use of genetic testing as a screening tool for the disease in the general population. However, a recent study suggests that less than 1% of subjects who are homozygous for the gene mutations will go on to develop the full blown disease of hereditary hemochromatosis, historically termed "bronzed diabetes." The study also suggests that homozygotes have no higher risk of mortality or of any clinically significant morbidity than normal control subjects. This conclusion contradicts earlier findings that linked iron overload and HFE mutations to a number of devastating diseases, including cardiovascular disease, diabetes, and cancer. PMID- 12638465 TI - They've entered the starting gate ... they're off! PMID- 12638464 TI - [The 3 spirits of humanity. So-called spiritual science in physiology of early modern times]. PMID- 12638466 TI - Who will take care of the patients? PMID- 12638467 TI - Childhood immunization: national, state rates still short of goal. PMID- 12638468 TI - MR cholangiopancreatography: a pictorial essay. PMID- 12638469 TI - A review of anthrax in the workplace. PMID- 12638470 TI - [Oral anticoagulation with vitamin K antagonists]. AB - Coumarins are broadly employed in clinical practice and should therefore belong to the group of 'well known' drugs. The present review will deal with some concepts of anticoagulant therapy: the role of vitamin K in the synthesis of functional coagulation factors, some clinically relevant pharmacokinetic aspects of coumarins, the management of oral anticoagulant therapy with special emphasis on the laboratory monitoring, and the most frequent complication, bleeding. PMID- 12638471 TI - [Heparins]. AB - Unfractionated heparin (UH) and low-molecular-weight heparins (LMWH) are antithrombotic drugs covering virtually all indications requiring immediately effective anticoagulation. For prevention and treatment of venous thromboembolism (VTE) UH have mainly been replaced by LMWH due to their practical usefulness (one or two subcutaneous daily doses without laboratory test for dose adjustment) and their more favourable risk-benefit profile. With respect to arterial occlusions this statement is also valid for unstable angina pectoris. The risk to develop heparin-induced thrombocytopenia (HIT) appears to be ten times lower with LMWH. In hospitals the use of UH is reserved for complex cases with high bleeding risk and the necessity to interrupt heparin effects rapidly. Therapeutic doses of UH are monitored by the classical coagulation assays, activated partial thromboplastin time (aPTT) and thrombin time (TT), but also by automated chromogenic tests of anti-factor Xa activity. There are no prospective studies directly comparing the efficiency of these three approaches. Disagreements between tests are not rare in individual cases. However, dosing recommendations for UH treatment of VTE (intravenous bolus of 80 IU/kg, followed by 18 IU/kg/h infusion) and average doses used are concordant. 0.30-0.70 anti-Xa IU/ml are considered as therapeutic range for UF infusion. Higher UH activities required during extracorporeal circulation in heart surgery or during coronary angioplasty are usually guided by bedside ACT (activated clotting time). For LMWH tests of anti-Xa activities may only be necessary during weight adjusted treatment of pregnant women, children, or cases with reduced kidney function (glomerular filtration rate < 30 ml/min.) or increased bleeding risk. Expected anti-Xa activities are 0.5-1.1 IU/ml and 1.0-2.0 IU/ml 4 hours after subcutaneous LMWH for dosing intervals of 12 hours and 24 hours respectively. PMID- 12638472 TI - [Antiaggregation: aspirin]. AB - Many randomized trials have shown aspirin as an effective antiplatelet drug for the secondary prevention of cardiovascular events. The NNT (number needed to treat) to prevent 1 vascular event is about 25. The NNH (number needed to harm) inducing one cerebral bleeding is about 1'000, to provoke one severe extracerebral bleeding about 100-200. The primary prevention can be recommended only for high risk patients for cardiovascular events (annual risk of 1-1.5% or more), calculated on the basis of the Framingham data, the Sheffield tables or in analysis of U.S. Preventive Services Task Force. The mechanisms of action, interactions and the "aspirin-resistance" are briefly discussed. PMID- 12638473 TI - [New anticoagulants -- their clinical significance]. AB - Heparin and Vitamin K antagonists have been the only available anticoagulants for several decades. Their use has lead to significant achievements in all fields of medicine despite various shortcomings and bleeding complications. With the objective of an improved benefit-/risk ratio selective inhibitors of factor Xa (Fondaparinux) and factor IIa (Ximelagatran) have been developed. Ximelagatran can also be orally administered. The results obtained from various clinical trials with these compounds are extremely encouraging. Thus, a significant improvement of antithrombotic treatment may be expected by their future use in the clinical and out-patient setting. PMID- 12638475 TI - [Anticoagulation and antiaggregation in cardiac patients]. AB - Aspirin treatment for primary prevention is safe and useful at an annual coronary event risk > or = 1.5%. Both aspirin and clopidogrel reduce the rate of cardiovascular events in patients with coronary disease. Clopidogrel in addition to aspirin further reduces cardiovascular events, but is associated with and increased bleeding risk. Recent studies in patients with myocardial infarction suggest that treatment with either coumadin or with coumadin and aspirin are both at least as effective than treatment with aspirin alone. Thromboembolism and bleeding during therapeutic anticoagulation are the major chronic risks for patients with native valvular heart disease and mechanical prosthetic valves. The recommendations for the prevention of thromboembolic events and bleeding complications are discussed and recommended intensity of antithrombotic therapy are outlined. Key points of the guidelines for managing patients with atrial fibrillation are summarised. PMID- 12638474 TI - [New antiaggregants and their importance for the practitioner]. AB - Since the introduction of Aspirin in the 1950s many substances have been introduced with an antiplatelet effect. Only few have been proved to be superior to aspirin in some clinical settings. Ticlopidin and its biochemical analog clopidogrel block the ADP-induced platelet aggregation. Clopidogrel having a better profile in terms of adverse events has been established over ticlopidin. It can be used for secondary prevention in patients with stroke, myocardial infarction or peripheral arterial disease instead of aspirin, certainly in cases with aspirin intolerance or with relapsing cardiovascular event under treatment with aspirin. For a general substitution of aspirin through clopidogrel hard evidence is still lacking. The combination aspirin + clopidogrel has been proved superior to aspirin in the treatment of patients with unstable angina or myocardial infarction without ST-segment elevation. For an extension of this indication to other clinical entities further evidence is needed. GPIIb/IIIa blockers are a well established treatment modality in patients undergoing coronary catheter interventions. Oral GPIIb/IIIa blockers, probably because of their pharmacokinetic profile, have proved to be insufficient in protecting from cardiovascular events compared to aspirin. Practical aspects concerning use of the newer antiplatelet agents during pregnancy, preoperatively and in spinal anesthesia are discussed. PMID- 12638476 TI - [Anticoagulation and antiaggregation in neurological patients]. AB - Aspirin is the drug of choice in most patients with acute stroke, if thrombolysis is contraindicated. Heparin is only used in acute stroke due to cerebral venous thrombosis, extracranial carotid or vertebral artery dissection and cardiac emboli with high risk of recurrence. In the prevention of recurrent stroke in patients with a noncardioembolic ischemic stroke antiplatelet agents are used. Aspirin is the first-line agent. Clopidogrel or a combination aspirin/dipyridamol are recommended for patients with several risk factors or recurrent cerebrovascular events. Warfarin has demonstrated a clear efficacy in stroke prevention in patients with atrial fibrillation, cerebral venous thrombosis and antiphospholipid antibody syndrome. Other, less well established possible indications for warfarin in the secondary prevention of stroke are symptomatic intracranial artery stenosis, large aortic atheroma, extracranial carotid or vertebral artery dissection and patent foramen ovale. PMID- 12638477 TI - [Anticoagulation and antiaggregation in patients with peripheral arterial occlusive diseases]. AB - Peripheral vascular occlusive disease (PAOD) is frequently seen in patients suffering from coronary heart or cerebrovascular disease and is, considered as a prognostic predictor for the morbidity and mortality of this patient group. Thus, secondary antithrombotic and antiplatelet prophylaxis in these patients is not limited to achievement of long-term patency of the revascularized or recanalized arterial segment, but plays as well a pivotal role for the prevention of myocardial infarction and stroke. Generally, claudicants as well as patients undergoing percutaneous transluminal angioplasty (PTA), supragenicular femoro popliteal artificial bypass surgery, aortofemoral, iliaco-femoral unilateral bypass, or aortobifemoral Y-graft implantation with unimpaired arterial outflow are treated life-long with low dose acetylsalicylic acid (ASA) 75-250 mg. On the other hand, those undergoing axillo-femoral, femoro-femoral crossover, aorto profundal or femoro-popliteal infragenicular and femoro-distal venous bypass surgery should be treated with vitamin K antagonists. The role of Clopidogrel in secondary prevention after peripheral revascularization and recanalization still needs to be defined. PMID- 12638478 TI - [Anticoagulation in patients with venous thromboembolism]. AB - Deep venous thrombosis and pulmonary embolism are considered to be two variants of one disease--'venous thromboembolism'. Pathogenesis, therapy and prognosis of these both entities are very similar and therefore the term 'venous thromboembolism' has been used in recent literature. The cornerstone of therapy is anticoagulation and initially consists of heparin for at least five days. Because of pharmacokinetic advantages low molecular weight heparins are the therapy of choice. They are as efficient and save as unfractionated heparins and allow weight-adapted dosing with daily subcutaneous injections in most patients. Low molecular weight heparins do not require regularly laboratory monitoring with few exceptions, e.g. renal failure. Therefore outpatient treatment of deep venous thrombosis is possible in most patients. Although there are promising data about outpatient treatment of pulmonary embolism, this is still being studied and can not be recommended outside clinical trials. Introduction of coumarin therapy for venous thromboembolism should be started on day 1 of diagnosis, keeping the total duration of heparin therapy at no more than five days and therefore minimizing the incidence of heparin-induced thrombocytopenia. Evidence from multiple studies indicates that effective coumarin therapy in venous thromboembolism is usually reflected by an INR of 2.0 to 3.0. In patients with massive and hemodynamically relevant thromboembolism alternative therapeutic approaches such as thrombolytic therapy, thrombectomy or insertion of intravenous filters may be useful. Adequately fit compression stockings can reduce the risk of post-thrombotic syndrome after deep venous thrombosis. PMID- 12638479 TI - [Anticoagulation and antiaggregation in pediatrics]. AB - The hemostatic system consisting of primary and secondary hemostasis as well as fibrinolysis is an evolving process throughout life with important qualitative and quantitative differences between children and adults. Moreover, the different age groups of childhood differ in many aspects of their hemostatic system. These differences reflect an adaptation to the young age rather than immaturity. Knowledge of the pediatric characteristics of the hemostatic system is the basis to diagnose, prevent and treat thromboembolic disorders. The achievements in intensive care medicine caused a higher incidence of thromboembolic events and, together with an increased knowledge of the pathophysiology, promoted hemostaseology as a pediatric subspecialty. The management of thromboembolic events with anticoagulants and platelet function inhibitors was first established in adult patients and many evidence based guidelines are achieved. However, clinical trials in children are rare due to the less developed network of the young pediatric subspecialty, and to the need of international cooperation because of small patient numbers. Based on this, experience made in adults is often extrapolated to similar situations in childhood neglecting the differences of the pediatric aspects of the hemostatic system, but also simply neglecting the child as a patient. The management of children with defects of the hemostatic system is therefore characterized by controversies that are further fueled by a lack of data. There is, however, a beginning of international activities with clinical trials and attempts of developing evidence based guidelines, or guidelines clearly stating where evidence is lacking. This activity should be supported by the international community of pediatricians who promote progress in knowledge and management of pediatric hemostatic system disorders. PMID- 12638480 TI - [Anticoagulation and antiaggregation during pregnancy]. AB - For haemostatic and circulatory reasons pregnancy is associated with an about 6 fold relative increase of thrombotic risk which is further raised by additional risk factors, such as history of thrombosis or acquired and hereditary thrombophilia, respectively. Recently, the thrombophilias have been revealed as risk factors for severe preeclampsia, abruptio placentae, fetal growth retardation, abortion and still birth as well. Thus, there are several situations in which the question for the need of antithrombotic medication is raised in a pregnant woman, either for therapy of acute thromboembolism or for prophylaxis of thrombosis and obstetrical complications, respectively. While acute thromboembolism has to be treated in every case, indication for prophylaxis has to be weighed in the light of the individual risk profile of the pregnant woman and the potential side effects and inconvenience of the medication. The first part of this article deals with the pregnancy related problems of coumarins, heparins and aspirin and demonstrates that the low molecular weight heparins are the anticoagulants of choice for most indications in pregnancy. The second part of this overview shows in which specific situations and how the antithrombotic medications mentioned above are used in pregnancy. PMID- 12638481 TI - [Patient self-management of oral anticoagulation]. AB - Self management of oral anticoagulation (OAC) has been demonstrated to improve the quality of oral anticoagulation as assessed by the percentage of INR values within the individual target range. There were tendentially less bleeding and thromboembolic complications in the group with self management of OAC. The capillary INR values of the only available monitor in Switzerland (CoaguChek, Roche Diagnostics, AG) correlated good with venous INR. Thus, self management of OAC may be a valuable alternative to conventional anticoagulation in selected and motivated patients on long term anticoagulation. PMID- 12638482 TI - [Clinical problems with oral anticoagulation -- 3 case reports]. AB - Two patients with severe bleeding complications under oral anticoagulant treatment are presented, in one case caused by pharmacokinetic drug interference (phenylbutazone), in the other by genetic predisposition to bleeding induced by coumarin anticoagulants. Another patient with decreasing INR due to drug interference (rifampicin) is presented as well. The possibility of drug interferences with coumarin anticoagulants has to be anticipated, whenever the medication of an orally anticoagulated patient is changed. A founder mutation of the factor IX propeptide constitutes a genetic predisposition to bleeding in patients put on coumarins. Its presence should be excluded in any patient suffering from hemorrhagic complications after starting anticoagulation when INR values are in the target range. PMID- 12638483 TI - [Mikhail Semenovich Lisitsyn (1891-1961)]. PMID- 12638484 TI - [Patients before and after surgery: starvation or adequate nutrition]. AB - The complete or partial starvation is the state often observed in surgical patients, especially before and after large surgical interventions on the organs of the digestion system. The inadequate nutrition is often thought to be the leading risk factor of postoperative complications and lethality. Of principal significance in prognosis and outcome of the disease in surgical patients is the timely and valuable nutritional maintenance (peroral, enteral, parenteral). At the present time about 100 medicines are used for enteral and parenteral nutrition of the patients. PMID- 12638486 TI - [Minimally invasive technologies in the treatment of cholelithiasis in high surgical risk patients]. AB - Using minimally invasive methods of sanitation of bile ducts in 456 patients with high operative risk reduced the number of postoperative complications to 11%, lethality to 1.3%. Two-stage treatment with surgical intervention by means of laparotomy was necessary in 124 patients. In this group of patients the frequency of complications was 32.3%, and lethality--4.8%. PMID- 12638485 TI - [Ganglionic blockers in abdominal surgery: facts and hypotheses. 2.Clinical, pathophysiological and pharmacological aspects of ganglionic blocker use]. AB - The authors discuss facts and hypotheses on the effects of benzohexonium upon the motor activity of the intestine and the significance of N-cholinolytics for prophylactics and treatment of postoperative pareses of the gastrointestinal tract. The ganglioblockers possess antistress effect, reduce the degree of pathological vegetative reactions and facilitate realization of the mechanisms of selfregulation of functions of the small and large intestine. Using benzohexonium during operation and in the first days after it makes the intestinal pareses less frequent. N-cholinolytics however do not have a considerable stimulating influence on the contracting activity of the gastrointestinal tract that accounts for their not high effectiveness in treatment of early functional motor evacuatory disorders. The points of action of gangliolytics, those at the level of the intestinal wall included, can not be considered to be completely established, as well as the mechanisms of their indirect effect. The ganglionic blockade should be considered as the basic method of prophylactics of the postoperative paresis of the intestine. PMID- 12638487 TI - [Comparative characteristics of octreotide and famotidine in the treatment of acute pancreatitis]. AB - Antisecretory therapy is an important and integral component of complex therapy of acute pancreatitis. The clinical estimation of efficiency of various antisecterory preparations, sandostatin and famotidin (quamatel) was made in treatment of acute pancreatitis at an early stage of the disease. The results of the research have shown that the efficiency of sandostatin and famotidin (quamatel) in treatment of not severe (mild and medium) forms of acute pancreatitis is approximately identical. But the application of famotidin (quamatel) is economically more favorable (8.3 times cheaper than sandostatin). In treatment of severe acute pancreatitis of paramount importance are the terms of the beginning of antisecretory therapy, the optimum term being the first day of the disease. The preparation of choice is sandostatin, the preparation of reserve--famotidin (quamotel). The duration of a medical course of treatment is the first three days from the beginning of the disease. The application of antisecretory preparations in question is less effective when used in later terms. PMID- 12638488 TI - [Minimally invasive technologies in the treatment of severe forms of acute pancreatitis at various periods of the disease]. AB - The choice of the optimum technique of the sanitation procedure in treatment of acute necrotizing pancreatitis (ANP) is now one of the most disputable problems. Clinical estimation of the efficiency of various operative techniques of treatment of ANP at various stages of disease has been made. In the aseptic phase laparoscopic decompression of the pancreas is indicated when the patient has evident hemorrhagic parapancreatitis. In the phase of septic sequestration of ANP the optimum method of minimally invasive surgical intervention is considered to be minilaparotomy which is expedient for abscesses of small volume, lipid abscesses of any volume with the minimal content of necrotizing tissues as the first step of sanitation in critical patient. High quantity of the necrotizing tissues in the zone of the destructive focus requires traditional laparotomy of the abscess under conditions of preventive maintenance of the endotoxic shock. PMID- 12638489 TI - [Diagnosis and choice of the method of surgical treatment of pancreatic cysts using ultrasonic examination]. AB - A more than 10 year experience of using operative interventions under the control of ultrasound has shown that in order to solve the task of improving the results of surgical treatment of cysts of the pancreatic gland it is necessary to improve a number of diagnostic and organizational measures allowing to perform the early and effective diagnosis of the cysts and their timely treatment. The percutaneous puncture-drainage operations with sclerotherapy are indisputably considered to be highly effective in treatment of cysts of the pancreas. In most patients, however, the clinical and ultrasonic signs of pancreatitis are still retained. These patients need permanent dynamic observation and conservative therapy. PMID- 12638490 TI - [Lymphotropic immune correction with myelopid in the postoperative period in patients with cancer of the colon and rectum]. AB - An analysis of surgical treatment of 142 patients with cancer of the colon and rectum included 32 emergency cases and 110 scheduled operations. Standard surgical methods only were used in 94 patients, the lymphotropic immunocorrection with myelopidum in the postoperative period was made in 48 patients. The use of lymphotropic immunocorrection was shown to considerably improve the immunological parameters and to decrease postoperative lethality. PMID- 12638491 TI - [Temporary prosthetics of limb arteries in the acute period of shock-inducing trauma]. AB - A retrospective analysis of treatment of 450 injured with traumas of the extremity vessels was made. These patients were subjected to reconstructive operations. In a considerable part of the patients there appeared thrombosis of the operated vessel. Experiments in dogs have shown that prophylactics of temporary prosthesis and injured artery should be based on the method of active treatment in order to eliminate the posttraumatic spasm of the arteries. PMID- 12638492 TI - [Use of immunomodulator derinat in the treatment of patients with surgical sepsis in traumatic shock]. AB - The reparative effects of Derinat were studied in 15 patients with traumatic shock whose state was complicated by the development of sepsis. The immunomodulating effect of the drug lies in increasing the number of lymphocytes, reducing the total number of granulocytes and in elevation of the number of functionally valuable cells in them. Using Derinat was shown to facilitate the restoration of the initially reduced number of blood erythrocytes. The maximum effect of the drug was observed within 3-4 days. Readministrations of Derinat should be made twice a week. PMID- 12638493 TI - [Clinico-endoscopic classification of acute anastomositis after resection of the stomach]. AB - The authors have analyzed the results of resection of the stomach in 106 patients with a complicated course of ulcer disease of the stomach and duodenum. Based on a complex investigation of regeneration of the gastrointestinal anastomoses formed by different methods and a comparison of the endoscopic picture with the clinical course of the early postoperative period the authors propose a clinico endoscopic classification of acute postgastroresectional anastomosites. Comparison of the frequency and severity of the anastomosites has shown advantages of a one-raw seroserous musculo-submucous suture in the resectional surgery of the stomach when forming gastro-intestinal anastomoses. The classification of acute anastomosites allows the timely detection and correction of the operation complications. PMID- 12638494 TI - [Possibilities of x-ray computed tomography in the diagnosis and treatment of suppurative complications of acute pancreatitis]. AB - The authors have analyzed results of the roentgen computed examination of 73 patients with acute pancreatitis and its complications. The potentialities of roentgen computed tomography (RCT) in diagnosing are shown as well as computed semiotics of acute pancreatitis and its complications. The authors consider that RCT makes the diagnosis substantially more exact and enables the complications to be detected much earlier. Systematization of the signs detected by RCT in acute pancreatitis in combination with the clinical symptoms gives the information necessary for correct strategy of surgical treatment. PMID- 12638495 TI - [Diagnosis and treatment of pseudoaneurysms after reconstructive interventions with the use of synthetic prostheses]. AB - The author analyses an experience with treatment of 43 patients with false aneurysms of anastomoses after reconstructions in the aorto-femoral zone (5.8% of all the patients operated on). A combination with an infection of the prostheses was registered in 9 patients, two of them had aneurysms of the proximal aorto prosthetic anastomosis. All the patients were operated upon. In three patients with aorto-prosthetic aneurysm resection of the proximal anastomosis was followed by reprosthesing and formation of a new aorto-prosthetic anastomosis. In combination with infection, the explants were completely ablated, the abdominal part of the aorta was ligated. The extraanatomical bifurcational axillary-femoral shunting was fulfilled with good results. When reconstructing the distal anastomoses the aneurysm was resected and the prosthesis was elongated with a linear insertion followed by the formation of a new distal anastomosis. The main blood flow in the extremity was obtained in all the patients. Lethality was 2.3%. PMID- 12638496 TI - [Use of perfluorane in surgical interventions on hollow organs under conditions of hemorrhagic shock (an experimental study)]. AB - Experiments in dogs have shown expediency of using perfluorane for prevention of incompetence of anastomosis sutures when the formation of intestinal anastomosis occurs under conditions of massive blood loss. PMID- 12638497 TI - [Experimental bases of a new method of epidural anesthesia in patients with closed thoracic injury]. AB - The thoracic epidural block is the most effective method of anesthesia in patients with a closed mechanical trauma of the chest. But this method is associated with a risk of damage of the spinal cord. So it can be used but by highly qualified specialists. The authors have worked out a method of anesthesia of the thoracic segments based on the phenomenon of hydrolock. It is more simple, safe, easy for any specialist able to use the technique of routine lumbar epidural block. PMID- 12638498 TI - [Diagnosis and intensive therapy of thermal and inhalation trauma]. AB - The authors made an analysis of an experience with treatment of patients with a combined thermal inhalation injury in the Burn Center of the St. Petersburg Research Institute of emergency medical care named after I.I. Dzhanelidze for the recent years. A prognostic model with respect to the patient's life was worked out on the basis of complex estimation of the data of the depth and square of the injury of skin, severity of the inhalation trauma and age of the casualty. The authors propose a scale of estimation of the severity of the inhalation trauma with special reference to the probable outcome which allows a differentiated approach to choice of the treatment policy that finally favors reduction of lethality in the group under analysis. PMID- 12638499 TI - [Modified laparoscopic fundoplication in the treatment of gastroesophageal reflux]. AB - The results of laparoscopic fundoplication in patients with the gastroesophageal reflux disease are described. Operations were performed by the classical Dor method (19 patients) and by a modified method (35 patients). An analysis of results of the operation has shown effectiveness and safety of using the modified method of laparoscopic fundoplication. PMID- 12638500 TI - [Use of polypropylene mesh in large and giant hernias of the anterior abdominal wall]. AB - An experience with treatment of 42 patients with large and giant hernias of the anterior abdominal wall is described. The treatment used plasty with a polypropylene net by the method used in the clinic. Long-term results were followed up in 36 patients during the period from 6 months to 5 years. Recurrences of hernias were not noted. PMID- 12638501 TI - [Rational surgical strategy in urgent non-gravidic metrorrhagia]. AB - The authors made an analysis of aspects of using the diagnostic and operative hysteroscopy in diagnosis and treatment of urgent nongravidary metarrhagias (UNM) under conditions of the department of urgent gynecology of the multiprophile hospital. The authors have analyzed the causes of the development of UNM, revealed high incidence of adenomyosis (44.3%). PMID- 12638502 TI - [Intrahepatic cholelithiasis after cholecystectomy]. PMID- 12638503 TI - [Pylephlebitic abscesses of the liver with favorable short- and long-term results of the treatment]. PMID- 12638504 TI - [Rare localization of a pancreatic cyst in the stomach wall]. PMID- 12638505 TI - [Ectopy of the pancreatic tissue in the jejunum complicated by hemorrhage]. PMID- 12638506 TI - [Primary multiple carcinoid of the rectum]. PMID- 12638507 TI - [Surgical reduction of the lung volume as a method of treatment of respiratory insufficiency in patients with diffuse emphysema. II]. PMID- 12638508 TI - [Treatment of peritoneal carcinomatosis of non-gynecological origin]. PMID- 12638509 TI - [Cytoreductive surgery as an alternative to palliative operations in oncology (the model of treatment of stage IV colorectal cancer)]. AB - Based on an analysis of treatment of patients with colorectal cancer of the IV stage in two groups of patients by the method of cytoreductive technology (58 patients) compared with a group (64 patients) treated using palliate operations, the author makes a conclusion that cytoreductive operations are an alternative to palliative interventions since they give longer survival and save quality of life. The adjuvant immunochemotherapy is thought to be a necessary component of cytoreductive operations. PMID- 12638510 TI - [Current questions of diagnosis and surgical treatment of metastatic cancer of the liver (materials of the 3d Russian-German symposium, Moscow, June 5-6, 2001)]. PMID- 12638511 TI - [The 9th Conference of Surgeons-Hepatologists of Russia and CIS (St Petersburg, May 16-17, 2002)]. PMID- 12638512 TI - Do stimulants for ADHD lead to abuse later on? PMID- 12638513 TI - Synthetic pillows and wheezing in childhood. PMID- 12638514 TI - Autism--is it really increasing? PMID- 12638515 TI - Backpack injuries in children--not what you think. PMID- 12638516 TI - Can "we just know" that a child was exposed to cocaine before birth? PMID- 12638517 TI - Product recalls. Recalls: children's soap making kits...wooden toy vehicles...talking dolls...and stuffed bunny toys. PMID- 12638518 TI - Cervical cytology screening. PMID- 12638519 TI - Aberrant receptor-mediated Cushing's syndrome. AB - Multiple alterations of G-protein-coupled receptors and G-proteins regulating intracellular transduction signal have been described in endocrine tumours. In Cushing's syndrome, aberrant or 'illicit' expression of membrane receptors (mainly G-protein-coupled receptors) has been observed in adrenal adenomas and adrenocorticotropic hormone (ACTH)-independent macronodular bilateral adrenal hyperplasia. The best characterized example to date is the aberrant expression of the gastric inhibitory polypeptide receptor that causes 'food-dependent hypercortisolism'. Aberrant expression of the luteinizing hormone, 2-adrenergic, interleukin receptors have also been reported. The level of expression of the vasopressin V1a receptor correlates with the direct (ACTH-independent) cortisol response to vasopressin. PMID- 12638520 TI - Adrenoleukodystrophy: the most frequent genetic cause of Addison's disease. PMID- 12638521 TI - Williams-Beuren syndrome: a model of recurrent genomic mutation. AB - Williams-Beuren syndrome is a segmental aneusomy syndrome with manifestations affecting the vascular, connective tissue, endocrine and central nervous systems. Most patients show a similar heterozygous approximately 1.5 Mb deletion at 7q11.23 that contains a number of reported genes. Deletion mapping in the few atypical patients with smaller deletions suggested that additive effects of haploinsufficiency for two or more genes might be necessary for the phenotype. Vascular stenoses are caused by haploinsufficiency at the elastin gene, while the genes responsible for the cognitive deficits are likely located at the telomeric edge of the deletion, including CYLN2 and GTF2I. Large region-specific segmental duplications predispose to misalignment and inter- or intrachromosomal unequal crossing-over causing the deletions. Atypical alleles at 7q11.23 such as inversions and deletions/insertions of large repeats, also generated through aberrant recombination between the local segmental duplications, are found in approximately 35% of transmitting parents. Genomic instability at 7q11.23 is directly related to the genomic structure of the region. PMID- 12638522 TI - Grave's disease 1835-2002. AB - This brief review describes the history of Graves' disease, starting with the original descriptions by Parry, Graves and von Basedow. The true aetiology of the disorder was uncovered in the 1950s and 1960s, based on the search for a novel thyroid stimulator which turned out to be an immunoglobulin G autoantibody. Assays for these thyroid stimulatory antibodies have been continually refined and their epitopes on the thyroid stimulating hormone receptor are increasingly well characterized. We also understand far more about the genetic and environmental susceptibility factors that predispose to disease, and even thyroid-associated ophthalmopathy has now been better defined as primarily a T-cell-mediated disease resulting from cytokine stimulation of orbital fibroblasts. These advances should improve treatment options for Graves' disease in the foreseeable future. PMID- 12638523 TI - Measurement and the newborn infant. AB - Measurement of newborn babies is widely regarded as being too inaccurate to justify its regular practice. It is common for infants to be weighed at birth and for no other measurements to be made. Although such assumptions are superficially correct, it is possible to train people to perform accurate measurements and for improved performance to be sustained. Accurate sequential measurements are possible and provide more information than single measurements. Detailed measurements show that postnatal growth may change rapidly and dramatically, particularly in preterm infants. Postnatal growth impairment is common in such infants and may be sustained. Limited evidence suggests that there may be a significant reduction in final stature. Preliminary data also suggest that many preterm infants may also show evidence of alterations in biochemical and physiological variables consistent with early programming and the potential for altered disease susceptibility in adult life. PMID- 12638524 TI - Natural growth in children born SGA with and without catch up growth. PMID- 12638525 TI - Polymorphisms and mutations in the GH-IGF axis in very short children born SGA due to IUGR, and the implications for the patients in childhood and adult life. PMID- 12638526 TI - Born small for gestational age: increased risk of type 2 diabetes, hypertension and hyperlipidaemia in adulthood. AB - The metabolic and cardiovascular complications associated with reduced fetal growth have been identified in the past 10 years. These include cardiovascular disease and the insulin resistance syndrome, comprising dyslipidaemia and impaired glucose tolerance or type 2 diabetes, and they appear to result from the initial development of insulin resistance. Although the mechanism underlying the development of insulin resistance associated with reduced fetal growth remains unclear, there is some evidence that adipose tissue plays a key role. Over the past decade, several hypotheses have been proposed to explain this unexpected association. Each points to either a detrimental fetal environment, genetic susceptibility or an interaction between the two. Although yet to be confirmed, the hypothesis suggesting that the association could be the consequence of genetic-environmental interactions is at present the most attractive. PMID- 12638527 TI - Effects of long-term growth hormone treatment on body composition, carbohydrate metabolism, blood pressure and lipids in short children born small for gestational age. PMID- 12638528 TI - Intellectual and psychological performance in males born small for gestational age. AB - All male singletons born without congenital malformations in Sweden between 1973 and 1978 and conscripted between January 1991 and January 1997 (n = 254426) were studied. Intellectual and psychological performance was tested at conscription. Males born small for gestational age (SGA) had lower results on both intellectual and psychological performance testing. Among males born SGA, low mean scores in both tests were constantly more common in those without catch-up growth than in those with catch-up growth. In conclusion, being born SGA is associated with an increased risk of subnormal intellectual and psychological performance. Catch-up growth is associated with a reduced risk of subnormal performance in males born SGA. PMID- 12638529 TI - Growth hormone treatment of children born small for gestational age. PMID- 12638531 TI - [Dear colleagues]. PMID- 12638530 TI - Long-term evolution of blood lipid profiles and glycemic levels in patients after spinal cord injury. AB - OBJECTIVE: To analyze the lipid and glycemic profiles in relation to age, time elapsed post injury and the level of injury in a group of patients with SCI. SUBJECTS: 2135 subjects with SCI were recruited during the annual routine health check-up carried out at the Institut Guttmann of Badalona, in Spain. The patients were classified according to the level of SCI and their age. METHODS: Serum lipid profiles and glycemic (glucose) levels were determined. Data was collected between 1996 and 2001, all biochemical analysis being performed by the same laboratory. RESULTS: Serum cholesterol levels show a statistically significant correlation with patient's age(P<0.001) and with time elapsed after injury (P<0.05). Differences can be observed between the values obtained from the different injury level groups (F=22.14; P<0.001). LDL-c shows a statistically significant difference between the four different age groups (F=3.51, P<0.05). There is an increase in plasma triglycerides related to the increase in age of the patients without observable differences between the injury level groups. The concentration of serum glucose increases with age (P<0.001) and also with the time elapsed after injury (P<0.05). CONCLUSION: There is a direct relationship between various biochemical parameters (total cholesterol, LDL-c and glucose) and the age of the patients, but not with the severity or time of evolution of the spinal cord injury. PMID- 12638532 TI - [Migraine therapy. new specific triptan frovatriptan--long acting]. PMID- 12638533 TI - [Information on preoperative analgesia. Acute pain therapy with rofecoxib]. PMID- 12638534 TI - [Triptane. Zolmitriptan--fast and effective in migraine]. PMID- 12638535 TI - [Guidelines -- safety net]. PMID- 12638536 TI - [Predictive criteria of endovascular treatment success in abdominal aortic aneurysms]. PMID- 12638537 TI - [Assisted death: open discussion in connection with pain therapy and palliative medicine applicable to Germany. Letter in response to W. Sohn in Der Sschmerz 16(2002): 150-152]. PMID- 12638538 TI - [Assisted death: open discussion in connection with pain therapy and palliative medicine applicable to germany. Letter in response to W. Sohn in Der Schmerz 16 (2002):150-2]. PMID- 12638539 TI - [Assisted death: open discussion in connection with pain therapy and palliative in Germany. Letter in response to W. Sohn in Der Schmerz 16(2002): 150-152]]. PMID- 12638540 TI - A voluntary moratorium? PMID- 12638541 TI - "Man's best friend": a new model system for cancer therapeutics? PMID- 12638542 TI - VP22 flips the switch on cell death. PMID- 12638543 TI - [Determination of plasma iron -- useful, superfluous, or misleading?]. PMID- 12638545 TI - [Role of bone histology in the determination of bone metabolism]. PMID- 12638546 TI - [Role of bone histology in the determination of bone metabolism]. PMID- 12638547 TI - [Sirolimus offers advantages for kidney transplant patients]. PMID- 12638550 TI - Infant and maternal health services in Ceylon, 1900-1948: imperialism or welfare? AB - This article contests a dominant contemporary view that colonial medicine was oppressive and detrimental to welfare; in particular, that infant and maternal welfare services were culturally hegemonic in their imposition of western practices and values on indigenous women. It does so by studying the development of these services in just one British colony--the 'model colony' of Ceylon from 1900 until independence. It shows how, at both a practical and a theoretical level, there was a direct policy transfer from the metropolitan centre to the colony. Moreover, the main justifications for the development of health and welfare services for women and children ran parallel to those used earlier in Britain. By 1948, these services were extensive in Ceylon and contributed to the fall in infant and maternal mortality rates at the end of the colonial period. It concludes by arguing that, just as in the West, these services were contradictory: they could both help maintain the exploitative State and enhance welfare. However, it is ultimately on their ability to improve health and welfare that they should be judged. PMID- 12638551 TI - The genesis of the notion of stages in oncology: the French Permanent Cancer Survey (1943-1952). AB - This article analyses the Enquete Permanente Cancer (EPC) (Permanent Cancer Survey), which organized the recording of cancers in France from 1943 onwards. This survey used a new classification of tumours and introduced the notion of stages. For its French authors, defining the different stages of the disease was primarily a question of methodological necessity: standardizing and giving relevance to data which until then has been disparate and scarce. However, by correlating the timing of the initial diagnosis of cancer with the length of asymptomatic periods after anti-cancer treatment, differentiated survival rates could be obtained. For each type of cancer, each stage could be shown to correspond to a greater or lesser probability of lenght of survival. This notion of stages, which was rapidly adopted on the international level as a tool for probabilistic reasoning, was not adopted for the purposes of prognosis in France until the 1960s. PMID- 12638552 TI - [T2-weighted MRI in a case with multiple cerebral cavernous malformations]. PMID- 12638553 TI - Making medicine indigenous: homeopathy in South India. AB - Historical studies of homeopathy in Europe and the USA have focused on practitioners' attempts to emphasize 'modern' and 'scientific' approaches. Studies of homeopathy in India have focused on a process of Indianization. Arguing against such unilineal trajectories, this paper situates homeopathy in South India within the context of shifting relations between 'scientific' and 'indigenous' systems of medicine. Three time periods are considered. From 1924 through 1934, homeopathy was singled out by Government of Madras officials as 'scientific', as contrasted with the 'indigenous' Ayurvedic, Siddha, and Unani systems of medicine. From 1947 through 1960, both 'indigenous' and 'scientific' interpretations of homeopathy were put forward by different factions. An honorary director of homeopathy proposed the Indianization of homeopathy, and its reconciliation with Ayurveda; this view conflicted with the Madras government's policy of expanding the 'scientific' medical curriculum of the Government College of Indigenous Medicine. It was not until the early 1970s that homeopathy was officially recognized in Tamilnadu State. By then, both homeopathy and Ayurveda had become conceptualized as non-Tamil, in contrast with promotion of the Tamil Siddha system of 'indigenous' medicine. Thus, constructs of 'indigenous' and 'scientific' systems of medicine are quite malleable with respect to homeopathy in South India. PMID- 12638554 TI - [Gene therapy of brain tumors--the current status and future prospect. An interview by Dr. Akira Teramoto]. PMID- 12638555 TI - [An autopsy case of familial periodic ataxia]. PMID- 12638556 TI - Major heart failure news from 2001. PMID- 12638558 TI - A randomized trial of splinting vs. surgery for carpal tunnel syndrome. PMID- 12638557 TI - 49th Annual Scientific Sessions of the American College of Cardiology. PMID- 12638559 TI - Cases from the Osler Medical Service at Johns Hopkins University. Sister Mary Joseph nodule. PMID- 12638560 TI - Active forgetting at the subcellular level. PMID- 12638561 TI - Disease and diversity in history. (Reviews of: Cartwright, FF, Biddiss, M. Disease and history. Gloucestershire: Sutton Publishing Limited, 2000; Rotberg RI, ed. Health and disease in human history. Cambridge, MA: MIT Press, 2000; Cliff, A., Haggett, P., Smallman-Raynor, M. Deciphering global epidemics: analytical approaches to the disease records of world cities, 1888-1912. Cambridge: Cambridge University Press, 1998; Riley, JC. Rising life expectancy: a global history. Cambridge: Cambridge University Press, 2001; McMichael, T. Human frontiers, environments and disease: past patterns, uncertain futures. Cambridge: Cambridge University Press, 2001). PMID- 12638562 TI - Heart failure update 2000. PMID- 12638563 TI - Prenatal stress has long term effects on pituitary development. PMID- 12638564 TI - Office-based and intraoperative ultrasound enhance surgeon's care of breast disease patients. PMID- 12638565 TI - Cerebrovascular reactivity to carbon dioxide in sepsis syndrome. AB - Cerebral dysfunction in sepsis is common in critically ill adults. However, little is known of the effects of sepsis on cerebral haemodynamics. We studied 12 sedated and ventilated patients in whom sepsis had been established for > 24 h. Transcranial Doppler measurements of the middle cerebral artery flow velocity were made at normocapnia, then hypocapnia (-1 kPa) and hypercapnia (+1 kPa). From these data, cerebrovascular reactivity to carbon dioxide was calculated. Variables indicating disease severity, systemic cardiovascular status and outcome were also recorded. We found significant changes in cerebrovascular reactivity to carbon dioxide. Only three of 12 patients had a cerebrovascular reactivity to carbon dioxide in the normal range; seven patients had a reduced cerebrovascular reactivity to carbon dioxide, whereas in two patients it was raised. In this smaD sample, we could not find any trend of association between altered cerebrovascular reactivity to carbon dioxide and severity of illness, cardiovascular status or outcome. This study suggests that established sepsis profoundly affects the vascular tone and reactivity, not only of the systemic circulation, but also of the cerebral vasculature. PMID- 12638566 TI - Prospective randomised double-blind comparative study of rocuronium and pancuronium in adult patients scheduled for elective 'fast-track' cardiac surgery involving hypothermic cardiopulmonary bypass. AB - The majority of cardiac anaesthetists in the UK use pancuronium for fast-track cardiac surgery. We compared the duration of action of pancuronium and rocuronium in patients undergoing fast-track hypothermic cardiopulmonary bypass and cardiac surgery. We determined whether patients would have had residual neuromuscular blockade at extubation. Twenty patients were randomly allocated to receive either pancuronium 0.1 mg x kg(-1) or rocuronium 1 mg x kg(-1). Neuromuscular function was assessed by acceleromyography; spontaneous recovery was evaluated by the train-of-four ratio measured at the adductor pollicis longus muscle. Median times to recover train-of-four ratio of 0.9 were 3 h 38 min for rocuronium and 7 h 52 min for pancuronium. The median difference in recovery times was 4 h 15 min (95% CI 2 h 30 min to 6 h 20 min; p = 0.0003 by Mann-Whitney test). None of the patients in the rocuronium group and seven of 10 patients in the pancuronium group had their extubations delayed because of residual neuromuscular blockade. Unless fast-track patients have neuromuscular function assessed before extubation, pancuronium should not be used. PMID- 12638567 TI - The impact of Acinetobacter baumannii in the intensive care unit. AB - Acinetobacter baumannii is a significant problem in critically ill patients. It is widespread, can colonise patients quickly and causes virulent infections. However, its overall impact on morbidity and mortality in the critically ill remains unmeasured. This study was designed to investigate A. baumannii colonisation and infection rates in a critically ill population over an 18-month period. Twenty-seven patients from a population of 347 were identified as having A. baumannii. Sixteen were colonised, whereas 11 were infected. Eleven of the 27 patients with A. baumannii died (41%). Of these, eight were colonised and three were infected. In the same period, 320 patients did not have A. baumannii and their mortality rate was 20% (n = 64). The mortality rate of patients with A. baumannii was significantly higher than that of patients without infection. PMID- 12638568 TI - The incidence of flushing on induction of anaesthesia in patients who blush easily. AB - Flushing (reddening and blotching of the skin) is seen frequently at induction of anaesthesia, is associated with anaesthetic agents such as thiopental and muscle relaxants, and is attributed to histamine release. The changes are generally confined to the neck and upper chest (the blush area). In conscious subjects, the mechanisms responsible for blushing in the same skin distribution are well defined and neurally mediated. We investigated the relationship between a history of blushing easily and flushing after intravenous induction o f anaesthesia. We interviewed 898 patients about to undergo general anaesthesia and asked them if they blushed easily. Anaesthesia was induced with thiopental followed by suxamethonium and/or alcuronium. We noted skin colour and the presence of a flush every 5 min for 20 min. Women reported blushing more than men (47% of women, compared with 33% of men, p < 0.001), and blushing was more common in young people (p < 0.001). In those women with a history of blushing, 32% flushed on induction of anaesthesia, compared with 6% of those who did not blush. In men, a flush was seen in 22% of those who blushed, and in 0.2% of those who did not. These differences in the frequency of flushing were significant (p < 0.001). In conclusion, flushing after induction of anaesthesia appears to be related to individual predisposition and may be neurally mediated. PMID- 12638569 TI - Effect of the Confidential Enquiries into Maternal Deaths on the use of Syntocinon at Caesarean section in the UK. AB - The recommended dose of Syntocinon used for uterine contraction at Caesarean section is5 units intravenously, given slowly. We conducted a survey of the use of Syntocinon at Caesarean section among 240 lead obstetric anaesthetists in the UK in 2001 and found that 155 (87%) of the 179 (75%) respondents gave 10 units, 77 of them (50%) by rapid bolus. The risks of Syntocinon, especially given by rapid injection, were highlighted in the report of the Confidential Enquiries into Maternal Deaths in the UK (1997-99), which was published at the end of 2001, and so the survey was repeated in 2002. Of the 256 forms sent, 198 completed replies were returned (77%); these indicated a dramatic change of practice: only 30 (15%) now gave 10 units and only 7 of these (23%) by a rapid injection. One hundred and sixty-seven respondents to the second survey (84%) stated they had changed their practice and 159 of these (95%) gave the Confidential Enquiries report as the main reason for change. These results highlight the importance of the Confidential Enquiries as a means of improving practice. PMID- 12638570 TI - European Society of Cardiology--XXIIIrd Congress. Stockholm, Sweden, September 1 5, 2001. PMID- 12638571 TI - [Comment on the article by Gross MW, et al. The effectiveness of radiation treatment in comparison with extracorporeal shockwave therapy (ESWT) in supraspinatus tendon syndrome. (Strahlenther Onkol 2002;178:314-20. No.6)]. PMID- 12638572 TI - The 50th annual scientific session of the American College of Cardiology. Orlando, FL, USA, March 18-21, 2001. PMID- 12638573 TI - European Society of Cardiology 2000. Amsterdam, The Netherlands, 26-30 August, 2000. PMID- 12638574 TI - 73rd Annual Scientific Sessions of the American Heart Association. New Orleans, LA, USA, November 12-15, 2000. PMID- 12638575 TI - Animal and growth development: regulatory mechanisms. Proceedings of a French Polish Symposium. Paris, France. September 25-26, 2002. PMID- 12638576 TI - Aspirin and mortality from coronary bypass surgery. PMID- 12638577 TI - Meeting report from the 74th annual scientific sessions of the American Heart Association (AHA). Anaheim, CA, USA, November 11-14, 2001. PMID- 12638578 TI - [The early work of Paracelsus in the fields of medicine, science and philosophy]. PMID- 12638579 TI - C-reactive protein in the prediction of cardiovascular events. PMID- 12638580 TI - C-reactive protein in the prediction of cardiovascular events. PMID- 12638581 TI - The anatomy and time course of semantic priming investigated by fMRI and ERPs. AB - We combined complementary non-invasive brain imaging techniques with behavioural measures to investigate the anatomy and time course of brain activity associated with semantic priming in a lexical-decision task. Participants viewed pairs of stimuli, and decided whether the second item was a real word or not. There were two variables, the semantic relationship between the prime and the target (related or unrelated) and the interval between the onset of prime and target (200 or 1000 ms), to vary the degree of semantic expectancy that was possible during task performance. Behavioural results replicated the well-established finding that identification of the target is facilitated by a preceding semantically related prime. Event-related functional magnetic resonance imaging (efMRI) identified two brain areas involved in the semantic-priming effect. Activity in the anterior medial temporal cortex was diminished when target words were primed by semantically related words, suggesting involvement of this brain region during active semantic association or integration. In contrast, activity in the left supramarginal gyrus in the temporal-parietal junction was enhanced for target words primed by semantically related words. Brain areas influenced by the interval between prime and target words, and by the interaction between word interval and semantic priming were also identified. A parallel experiment using event-related potentials (ERPs) unveiled a striking difference in the time course of semantic priming as a function of expectancy. In line with previous reports, the primary effect of semantic priming on ERPs was the attenuation of the N400 component, in both short- and long-interval conditions. However, the priming effect started significantly earlier in the long-interval condition. Activity in the anterior medial temporal cortex has previously been shown to contribute to the N400 component, a finding that links the priming results obtained with efMRI and ERP methods. PMID- 12638582 TI - Reviparin after leg injury requiring immobilization. PMID- 12638583 TI - Reviparin after leg injury requiring immobilization. PMID- 12638584 TI - Two registers of madness in Enlightenment Britain. Part 1. AB - Part 1 of this essay argues that in Augustan and Georgian England it was widely understood that madness could have two more-or-less distinct meanings. 'Moral' madness was the subject's own fault, and he/she remained accountable for actions commissioned under its effects. The delusional ideas arose in the mind, and by definition remained within the moral province of the individual. By contrast, in 'real' madness, the sufferer was the passive recipient of body-based sickness, and he/she was understood to be innocent, but paid for this exculpation of moral accountability by surrendering full personhood. Part 2 of the essay examines how Georgian churchmen viewed madness, and then returns to psychiatry proper for a closer look at how certain theories of madness were inflected by the problematics of insanity's conflicting meanings. Finally, a previously unrecognized Georgian psychiatric therapy, 'classical moral treatment', is described and put in the context of the better-known Tukean 'moral treatment'. PMID- 12638585 TI - Proceedings of the VIth International Potsdam Symposium on Tick-borne Diseases (IPS-VI). Berlin, Germany. April 26-27, 2001. PMID- 12638586 TI - Cyclin E in breast cancer. PMID- 12638587 TI - Cyclin E in breast cancer. PMID- 12638588 TI - Cyclin E in breast cancer. PMID- 12638589 TI - Skin ulcers misdiagnosed as pyoderma gangrenosum. PMID- 12638590 TI - Linked by pain. Injured and ill workers can find many kinds of help online. PMID- 12638591 TI - Biomarkers in nutritional epidemiology. AB - OBJECTIVE: To illustrate biomarkers of diet that can be used to validate estimates of dietary intake in the study of gene-environment interactions in complex diseases. DESIGN: Prospective cohort studies, studies of biomarkers where diet is carefully controlled. SETTING: Free-living individuals, volunteers in metabolic suites. SUBJECTS: Male and female human volunteers. RESULTS: Recent studies using biomarkers have demonstrated substantial differences in the extent of measurement error from those derived by comparison with other methods of dietary assessment. The interaction between nutritional and genetic factors has so far largely gone uninvestigated, but can be studied in epidemiological trials that include collections of biological material. Large sample sizes are required to study interactions, and these are made larger in the presence of measurement errors. CONCLUSIONS: Diet is of key importance in affecting the risk of most chronic diseases in man. Nutritional epidemiology provides the only direct approach to the quantification of risks. The introduction of biomarkers to calibrate the measurement error in dietary reports, and as additional measures of exposure, is a significant development in the effort to improve estimates of the magnitude of the contribution of diet in affecting individual disease risk within populations. The extent of measurement error has important implications for correction for regression dilution and for sample size. The collection of biological samples to improve and validate estimates of exposure, enhance the pursuit of scientific hypotheses, and enable gene-nutrient interactions to be studied, should become the routine in nutritional epidemiology. PMID- 12638592 TI - The use of food-frequency questionnaires for various purposes in China. AB - OBJECTIVE: To compare various food-frequency questionnaires (FFQs) used in nutritional studies in China for various purposes. DESIGN: In Study 1, a simplified FFQ with 17 questions on food was used in a large rural study. In Study 2, a questionnaire consisting of 84 questions on food consumption of 16 food categories was used in a study comparing dietary consumption data and various health indicators of elderly people in four geographical areas in China. In Study 3, a questionnaire with 149 items in 17 food categories is being validated by comparison with data obtained by repeated 24-hour recalls. SETTING: Study 1 was carried out in one southern site and a northern site in 1996 to 1997. Study 2 was conducted in four different geographical sites in 1998. The on-going study, Study 3, has been carried out in Jiangsu and Beijing since 1999. SUBJECTS: Study 1 included 12 234 rural Chinese adults aged 40 years. There were 546 elderly people in Study 2. Study 3 is collecting data from 300 healthy adults. RESULTS: The results of food consumption and nutrient intakes from Study 1 were comparable with those obtained from a previous household dietary survey, in which sensible correlations between diet and diseases were also found. In Study 2, the dietary data from the four geographical areas showed significant differences in food and nutrient intakes among the different areas. The validation of the new FFQ in Study 3 is still going on. CONCLUSION: The FFQ is a useful method for the collection of individual food consumption information. The above FFQ forms could be used in studies with different purposes, especially in studying the relationship between diet, nutrition and chronic diseases. PMID- 12638593 TI - Measuring household food security in poor Venezuelan households. AB - OBJECTIVE: To validate abbreviated methods that estimate food security level among poor communities in Caracas, Venezuela. DESIGN: Two independent cross sectional studies were undertaken to internally and externally validate simple quantitative/qualitative methods. The quantitative measure was constructed from data on household food availability, gathered using the list-recall method. It is a count of the foods that explain 85% or more of household energy availability. The qualitative measure is a score of female-perceived food insecurity level estimated with a modified 'hunger index', reflecting food resource constraints and hunger experiences within the home. Socio-economic and food behaviour data that may predict household food security (HFS) levels were gathered. The second study was repeated a year later to measure the impact of an increase in the minimum wage on HFS levels. SETTING: Two poor urban communities in Caracas, Venezuela. SUBJECTS: All households in both communities that complied with selection criteria(poor and very poor families that share food resources) and were willing to participate. The sample comprised 238 and 155 female household food managers in the two communities. RESULTS: In 1995, data from females in 238 urban poor households provided evidence for the overall validity of the method. Its application in 1997 to 155 households in the other community gave support to the external validity of the method. Measures were repeated in 1998 on 133 subjects of the above sample, when the minimum wage was increased by 23%. Evidence is presented showing the sensitivity of the method to changes in the determinants of HFS. Data analysed during these three periods suggest that the method can be simplified further by using the food diversity score instead of the quantitative measure since these variables correlate highly with one another(r > or = 2 0.854). CONCLUSIONS: This simple method is a valid and precise measure of food security among poor urban households in Caracas. Th equalitative/quantitative measures complement each other as they capture different dimensions of HFS. PMID- 12638594 TI - Biomarkers and the measurement of fatty acids. AB - OBJECTIVE: To review the various biomarkers of dietary intakes of fatty adds in human populations, their measurement, limitations and analytical considerations. DESIGN: Review of the literature. RESULTS: Although there is no good biomarker of intake of total fat, a number of alternatives exist for assessing the intakes of exogenously produced fatty acids that are consumed. Adipose tissue, erythrocyte membrane concentrations and serum or plasma levels can reflect prior intakes over the past few hours to the past few years. The concentrations of individual fatty acids in these media generally reflect relative levels, and are influenced by a number of factors. Although relatively expensive to analyse, a single analysis by gas chromatography or high-performance liquid chromatography provides information on multiple fatty acids, and is superior to attempting to measure specific fatty acids using traditional dietary assessment methods. CONCLUSIONS: Biomarkers of fatty acids that reflect long-term intake are available for nutritional epidemiology purposes. Analytical methods have become very accurate and able to detect and quantify smaller families, such as trans-fatty acids. PMID- 12638595 TI - Erotomania: a conceptual history. AB - Four historical convergences are identified in the history of erotomania. According to the first, which lasted from Classical times to the early eighteenth century, erotomania was a 'general disease caused by unrequited love'. According to the second, erotomania was a disease of 'excessive physical love (nymphomania)'; this view remained active well into the nineteenth century. The third convergence focuses on the view that erotomania is a form of 'mental disorder', and this was held throughout the twentieth century. The fourth and current definition (a development of the third stage) sees erotomania as a 'the delusional belief of being loved by someone else'. Differences between Anglo Saxon and French views cast doubt on the meaning or coherence of the much-abused English eponym 'de Clerambault syndrome'. Erotomania is a construct, a mirror reflecting Western views on spiritual and physical love, sex, and gender inequality and abuse. On account of this, it is unlikely that there will ever be a final, 'scientific' definition rendering erotomania into a 'natural kind' and making it susceptible to brain localization and biological treatment. PMID- 12638596 TI - Measuring food intake in studies of obesity. AB - The problem of how to measure habitual food intake in studies of obesity remains an enigma in nutritional research. The existence of obesity-specific underreporting was rather controversial until the advent of the doubly labelled water technique gave credence to previously anecdotal evidence that such a bias does in fact exist. This paper reviews a number of issues relevant to interpreting dietary data in studies involving obesity. Topics covered include: participation biases, normative biases,importance of matching method to study, selective underreporting, and a brief discussion of the potential implications of generalised and selective underreporting in analytical epidemiology. It is concluded that selective underreporting of certain food types by obese individuals would produce consequences in analytical epidemiological studies that are both unpredictable and complex. Since it is becoming increasingly acknowledged that selective reporting error does occur, it is important to emphasise that correction for energy intake is not sufficient to eliminate the biases from this type of error. This is true both for obesity-related selective reporting errors and more universal types of selective underreporting, e.g. foods of low social desirability. Additional research is urgently required to examine the consequences of this type of error. PMID- 12638597 TI - Health reform and change in the Asia-Pacific region. Opening address. PMID- 12638598 TI - Uses and limitations of statistical accounting for random error correlations, in the validation of dietary questionnaire assessments. AB - OBJECTIVE: To examine statistical models that account for correlation between random errors of different dietary assessment methods, in dietary validation studies. SETTING: In nutritional epidemiology, sub-studies on the accuracy of the dietary questionnaire measurements are used to correct for biases in relative risk estimates induced by dietary assessment errors. Generally, such validation studies are based on the comparison of questionnaire measurements (Q) with food consumption records or 24-hour diet recalls (R). In recent years, the statistical analysis of such studies has been formalized more in terms of statistical models. This made the need of crucial model assumptions more explicit. One key assumption is that random errors must be uncorrelated between measurements Q and R, as well as between replicate measurements R1 and R2 within the same individual. These assumptions may not hold in practice, however. Therefore, more complex statistical models have been proposed to validate measurements Q by simultaneous comparisons with measurements R plus a biomarker M, accounting for correlations between the random errors of Q and R. CONCLUSIONS: The more complex models accounting for random error correlations may work only for validation studies that include markers of diet based on physiological knowledge about the quantitative recovery, e.g. in urine, of specific elements such as nitrogen or potassium, or stable isotopes administered to the study subjects (e.g. the doubly labelled water method for assessment of energy expenditure). This type of marker, however, eliminates the problem of correlation of random errors between Q and R by simply taking the place of R, thus rendering complex statistical models unnecessary. PMID- 12638599 TI - Health reform and change in the Asia-Pacific region. Keynote address. PMID- 12638600 TI - Easy placement of open-coil springs. PMID- 12638601 TI - Proceedings of the 22nd Annual International Gravitational Physiology Meeting, 22 27 April, 2001, Budapest, Hungary. PMID- 12638602 TI - Changes in gene expression and signal transduction in microgravity. AB - Studies from space flights over the past three decades have demonstrated that basic physiological changes occur in humans during space flight. These changes include cephalic fluid shifts, loss of fluid and electrolytes, loss of muscle mass, space motion sickness, anemia, reduced immune response, and loss of calcium and mineralized bone. The cause of most of these manifestations is not known and until recently, the general approach was to investigate general systemic changes, not basic cellular responses to microgravity. This laboratory has recently studied gene growth and activation of normal osteoblasts (MC3T3-El) during spaceflight. Osteoblast cells were grown on glass coverslips and loaded in the Biorack plunger boxes. The osteoblasts were launched in a serum deprived state, activated in microgravity and collected in microgravity. The osteoblasts were examined for changes in gene expression and signal transduction. Approximately one day after growth activation significant changes were observed in gene expression in 0-G flight samples. Immediate early growth genes/growth factors cox 2, c-myc, bcl2, TGF beta1, bFGF and PCNA showed a significant diminished mRNA induction in microgravity FCS activated cells when compared to ground and 1-G flight controls. Cox-1 was not detected in any of the samples. There were no significant differences in the expression of reference gene mRNA between the ground, 0-G and 1-G samples. The data suggest that quiescent osteoblasts are slower to enter the cell cycle in microgravity and that the lack of gravity itself may be a significant factor in bone loss in spaceflight. Preliminary data from our STS 76 flight experiment support our hypothesis that a basic biological response occurs at the tissue, cellular, and molecular level in 0-G. Here we examine ground-based and space flown data to help us understand the mechanism of bone loss in microgravity. PMID- 12638603 TI - Expression of cell adhesion molecules and lymphocyte-endothelium interaction under simulated hypogravity in vitro. AB - Using histochemical staining and FACS-analysis we have studied the basal and TNF alpha induced expression of E-selectin, ICAM-1 and VCAM-1 in human umbilical vein endothelial cells (ECs) exposed to simulated hypogravity. Control ECs did not contain detectable amounts of E-selectin or VCAM-1 but were ICAM-1 positive. As soon as after 6-8 hrs of clinorotation at 5 RPM the cellular content of ICAM- 1 increased. Moreover, hypogravity potentiated the effect of inflammatory cytokines (TNF-alpha and IL-1) on ICAM-1 expression. No increase in E-selectin or VCAM-1 expression was observed in ECs exposed to hypogravity itself. However, hypogravity reduced E-selectin and VCAM-1 expression in cell cultures activated by cytokines, more visible at their low (5-10 U/ml) concentrations. Both, control and clinorotated ECs poorly supported spontaneous lymphocyte adhesion; the adhesion of PMA-activated leukocytes was 15-20-fold higher. The interaction of unstimulated lymphocytes with cytokine-activated endothelium was more noticeable but significantly lower in cultures exposed to hypogravity. Activated blood cells interacted with endothelium more effectively, particularly, under hypogravity. Obtained results suggest that EC adhesion molecule expression and endothelium lymphocyte interaction are altered under simulated hypogravity conditions in direction of increase of endotlielial adhesiveness for activated blood cells. PMID- 12638604 TI - Cytogenetic characteristic of osteogenic cells in vitro as perspective predictors of osteopenia under microgravity. AB - Mechanical stimulation of bone tissue determined by earth gravity is one of the main factors mediating the nature, rate and direction of functional adaptation of the bone system in the process of onto- and phylogenesis. Theoretically expected losses of bone mass under condition of mechanical load deficit under microgravity (osteopenia, osteoporosis) may become a factor that limits the duration of space flights. As a result of long-term studies some properties and regularities of change in human tissue after prolonged space flights (for 5-7 months) were established. PMID- 12638605 TI - Space bioreactors: their use, their future. PMID- 12638606 TI - Tissue engineering in space. AB - The purpose of this paper is to present the status of that part of the [Microgravity Application Program] project related to the study of cartilage formation from pig chondrocytes. The work carried out so far followed two lines: (i) chondrocytes were incubated for up to three weeks in the RPM; (ii) a module developed for in-vitro cartilage formation will be tested in a sounding rocket flight (MASER 9, November 2001). PMID- 12638607 TI - Effects of simulated microgravity on metabolic activities related to DNA damage and repair in lymphoblastoid cells. AB - We adopted a simple experimental framework to follow the dependence of structural aberrations and the modifications in selected metabolic processes correlated with the exposure of cells to microgravity. Alterations to the cellular metabolism induced by exposure to microgravity are evidentiated in the modification of PARP activity (strongly dependent to the presence of DNA damages and to the altered gene expression), in the modification of the repair ability and in the cell's energy homeostasis (NAD and ATP). Cells are exposed continuously to microgravity in a Random Positioning Machine (RPM) in complete medium for 48 hours. At the end of this period a part of these cells are immediately analysed for the parameters reported above and the remaining were furtherly incubated in standard laboratory conditions to document eventual defects during the phases of the recovery process. A part of cells, just after exposure to microgravity, were also subjected to treatment with a strong damaging agent, KBrO3, and these cells were subsequently analyzed. This final treatment was meant to amplify the eventual deficiencies experienced by microgravity-exposed cells in the DNA repair process also in dependence with the alterated metabolic conditions resulting after the exposure to microgravity. PMID- 12638608 TI - Simulated microgravity conditions affect poly(ADP-ribose) polymerase activity in cultured human lymphocytes. PMID- 12638609 TI - Enhancement of protease activity involved in modulation of cell mutability by microgravity stress in UVC-irradiated human cells. AB - It is an intriguing question whether gravity-changing stress modulates human cell mutability. To resolve this problem, it is necessary to determine the cellular events leading to modulation. We previously detected protease activation just after UV (UVC, principally 254 nm wavelength) irradiation followed by hypomutability in cultured human cells. We here investigated whether UV-activated protease activity is affected in human UVAP-1 cells exposed to gravity-changing stress prior to UV irradiation. PMID- 12638610 TI - The ultrastructure of Chlorella mitochondria and their energetic state under altered gravity. AB - This study looks at the predominance of the participation of the mitochondria in the energy supply of Chlorella cells. The localization of the Mg(2+)-activated ATPases were studied during the culture development under the geterotrophic conditions, and with an investigation of the mitochndria ultrastructure at the various stages of the culture development under clinorotation. It was found that the activity level of this enzyme is one main indice of the energy state of these organelles. PMID- 12638611 TI - Reproduction during spaceflight by plants in the family Brassicaceae. PMID- 12638612 TI - Gravicurvature loss, changes in ultrastructure and calcium balance of pea root statocytes treated with EGTA. AB - Calcium is known to play a key role in the regulation of a wide variety of cellular events in plants, in particular as a second messenger in the transduction of gravity signals in statocytes. In order to investigate the role of extracellular calcium in gravity perception, many investigators have used ethylene glycol-bis-(aminoethyl ether)-N,N'-tetraacetic acid (EGTA), whose main property is the ability to form stable water-soluble complex compounds with divalent metals, particularly with Ca2+ ions, that do not penetrate across plasma membrane into a cell. PMID- 12638613 TI - Orientation of root hair growth is influenced by simulated microgravity. AB - We have tried to investigate the mechanisms supporting the plagiotropic growth (growth in parallel to the Earth) of root hairs in simulated microgravity. Our strategy to understand the regulation of such type of growth depends upon the study of cytoskeleton topography and calcium ions distribution in root hairs both in control and simulated microgravity. PMID- 12638614 TI - The influence of the HGMF on mass-charge transfer in gravisensing cells. AB - The present work is focused on the influence of the high-gradient-magnetic field (HGMF) on spatial distribution of ion fluxes along the roots (a), cytoplasmic streaming (b), and the processes of plant cell growth connected with intracellular mass and charge transfer (c). PMID- 12638615 TI - Clinorotation [correction of Clinoratation] effects on the structural-functional response in potato minitubers. AB - It is established that high plant growth and development in microgravity occurred normal. However, the change of plant growth rate is accompanied by the change of carbohydrate metabolism in photosynthesized cells (Kordyum, 1997). The decrease of starch grain size in chloroplasts and the decrease of content cellulose in cell wall were revealed (Sytnik et al., 1984; Nedukha, 1996). The change carbohydrate metabolism in photosynthesized organs could influence on the growth of underground organs and content of storage carbohydrates in these organs. Therefore, the aim of our study was to investigate the long-term clinorotation influence on the formation, structure of potato minitubers and content of starch and sugars in minitubers. PMID- 12638616 TI - Adenilate pool and thylakoid ATP-synthase content in pea leaves under clinorotation. AB - It is known that plant resistance to stress factors is connected with energy metabolism. The energy stored in the process of photophosphorylation in the form of ATP is used then to support respiration, transpiration, organic compound synthesis, growth and development as well as to restore cell structure after its damage under extremal environmental factors. Transformation of light energy into chemical energy of ATP is catalyzed by thylakoid membrane enzymatic complex of ATPsynthase-CF1CF0. Its activity and amount in the thylakoid membrane depends on plant growth conditions. The aim of this work was investigation of clinorotation effect on light-induced dynamics of adenyl nucleotides (AMP, ADP and ATP) and estimation of CF1CF0 content in thylakoids of pea leaves grown under slow clinorotation and vertical control. PMID- 12638617 TI - Nucleolar structure and function under clinorotation. AB - A nucleolus is the dynamic part of a cell nucleus where rDNA transcription, rRNA precursor processing and transport are proceeding. Investigations of the influence of microgravity and clinorotation on the structure and function of nucleolus began more than thirty years ago but, unfortunately, now there are only some articles concerning the nucleolar peculiarities in altered gravity. Available data about alterations of nucleolar morphology and function under the influence of high and low temperature, hypoxia and UV-microbeam irradiation allow us to put intent attention to the study of the nucleolus reactions on clinorotation. Therefore, the aim of our work was to investigate the ultrastructural organization and functional activity of nucleoli in Lepidium sativum root meristematic cells under clinorotation in order to understand whether a nucleolus can react to gravity. PMID- 12638618 TI - Influence of clinorotation on some parameters of photosynthetic apparatus. AB - This study aimed to examine the electron transport rates in the thylakoids, isolated from leaves of pea plants grown under clinorotation and in vertical control, to measure the chlorophyll a/b (Chl a/b) ratio in such thylakoids and in photosystem I (PSI) particles isolated from them, to elucidate if there are any differences in changes of PS II activity in thylakoids and Chl a/b ratio in PS I particles under phosphorylation of polypeptides of thylakoid pigment-protein complexes. PMID- 12638619 TI - Effect of clinorotation on growth and pigment biosynthesis in etiolated barley plants. AB - Altered gravity conditions (hyper- or hypogravity) leads to changes in metabolic processes in living organisms (Kordyum, 1997). One important subject of plant space biology is the investigation of the effects of microgravity conditions on the development of the photosynthetic apparatus. The impact of microgravity on the photosynthetic apparatus of plants has been studied in a number of space flight experiments. Particularly, the Chl (a+b) content and Chl a/b ratio were determined in several experiments, however the results did not allow understanding how microgravity affects pigment apparatus of the plants since presented contradictory results [Laurinavichius et al. 1984; Volovik et al, 1999]. To elucidate how clinorotation affects pigment apparatus formation in etiolated plants, we studied morphological characteristics and pigment (chlorophyll and carotenoids) biosynthesis in the first period of greening of barley plants. PMID- 12638620 TI - The interaction of microgravity and ethylene on the ultrastructure cell and Ca2+ localization in soybean hook hypocotyl. AB - Calcium ions are secondary messenger in numerous cellular processes of plant grown at 1 g. Ca2+ are connected with oxygen atoms, of pectin carboxy groups and/or with H(+)-groups of protein (Roux and Slocum, 1982; Hepler and Wayne, 1985). The influence of altered gravity on the calcium balance in some cells is established. The increased synthesis of ethylene in plant grown in microgravity caused the change of the structural-functional organization of cell (Hensel and Iversen, 1980; Hilaire et al., 1996). Available data put the new question: how do high ethylene level and microgravity influence on the redistribution of Ca2+ in cell of seedling in early stage of growth? Therefore, the goal of our data was the comparable study of the cell ulltrastructure and localization of Ca2+ in hook hypocotyl of soybean seedling under interaction of microgravity and ethylene. PMID- 12638621 TI - Is there resetting of central venous pressure in microgravity? AB - In the early phase of the Space Shuttle program, NASA flight surgeons implemented a fluid-loading countermeasure in which astronauts were instructed to ingest eight 1-g salt tablets with 960 ml of water approximately 2 hours prior to reentry from space. This fluid loading regimen was intended to enhance orthostatic tolerance by replacing circulating plasma volume reduced during the space mission. Unfortunately, fluid loading failed to replace plasma volume in groundbased experiments and has proven minimally effective as a countermeasure against post-spaceflight orthostatic intolerance. In addition to the reduction of plasma volume, central venous pressure (CVP) is reduced during exposure to actual and groundbased analogs of microgravity. In the present study, we hypothesized that the reduction in CVP due to exposure to microgravity represents a resetting of the CVP operating point to a lower threshold. A lower CVP 'setpoint' might explain the failure of fluid loading to restore plasma volume. In order to test this hypothesis, we conducted an investigation in which we administered an acute volume load (stimulus) and measured responses in CVP, plasma volume and renal functions. If our hypothesis is true, we would expect the elevation in CVP induced by saline infusion to return to its pre-infusion levels in both HDT and upright control conditions despite lower vascular volume during HDT. In contrast to previous experiments, our approach is novel in that it provides information on alterations in CVP and vascular volume during HDT that are necessary for interpretation of the proposed CVP operating point resetting hypothesis. PMID- 12638622 TI - Cardiac, cerebral & lower limb hemodynamic changes, during HDT (5 days to 42 d) & space flights (7 days to 6 months). AB - The objective was to identify the major cardiovascular changes induced by exposure to real or simulated 0 g (spaceflights: 6 days, 14 d, 21 d, 25 d, 6 months; Head Down Tilt: 10 h, 4 d, 5 d, 7 d, 30 d, 42 d), with a minimum of countermeasure (Daily exercise in space, no exercise in HDT). PMID- 12638623 TI - The neurogenic constrictor response of isolated rat saphenous artery is reduced after 2-week tail suspension. AB - Under real or simulated microgravity conditions the control of arterial vascular tone is greatly disturbed. The low arterial vessel reactivity to sympathetic influences may be the cause of an increase in flow in hind limb skeletal muscles in tail-suspended (TS) rats. Our previous experiments with constant pressure perfusion of rat hind limb demonstrated the reduced vasoconstrictor responses to sympathetic nerve stimulation in TS rats. Responses to exogenous noradrenaline depended on the perfusion conditions. It is known that the vessels of various branching orders noticeably differ in nerve density and in sensitivity to vasoconstrictor agonists. So under neurogenic or exogenous noradrenaline influences the vascular resistance may be increased at different levels of vascular bed, thus making the data analysis seriously complicated. This uncertainty may be overcome by investigation of a single vessel isolated from hind limb vascular bed. The saphenous artery, a resistance artery with dense innervation, is a very convenient object for this purpose. Thus, this study was aimed at comparing the effects of 2-week tail suspension upon the constrictor responses of isolated saphenous artery to neurogenic and exogenous noradrenaline stimuli in rats. PMID- 12638624 TI - Average magnitude of heartbeat fluctuations in healthy subjects after exposure to 4 hours head down bed rest. AB - Orthostatic intolerance, induced by microgravity exposure, has been hypothesized to be related to a disorder of the autonomic control of the cardiovascular system. Non-invasive measurement of Heart Rate Variability (HRV) have been used as a valuable tool to characterize the ability of the autonomic system to modulate the cardiovascular function by analyzing the spontaneous fluctuations of arterial pressure and heart period on a beat-to-beat basis. Concerning this, conflicting results have been reported on the heart rate and blood pressure variability responses during exposure to microG. These differences seem to be due to different experimental designes used. Moreover, the different behavior of normal subjects in response to orthostatic stress after HD, i.e. Symptomatic (S) or Non Symptomatic (NS), could play some role in producing these discrepancies [correction of dicrepancies]. In this study we suggest the analysis of "magnitude" or "volatility" as a new method to study HRV variability. The volatility is a measure of the beat-to-beat excursion capability of the heart, regardless to its direction (positive or negative): the higher the volatility, the greater the excursion. The aim of the present study was to examine R-R volatility before and after 4 hours of HD -6 degrees in normal subjects. PMID- 12638625 TI - Effects of water depth on abdominal [correction of abdominails] aorta and inferior vena cava during standing in water. AB - During water immersion bradycardia and increase in stroke volume induce by changing in hydrostatic pressure. We hypothesised that the cardiac alterations with immersion are associates with an increase in venous return from lower body. Because the venous return reflects volume of capacitance vein e.g, inferior vena cava. Therefore, we clarified the effects of water immersion on the size in left ventricle, aorta, and vena cava. PMID- 12638626 TI - Blood pressure and heart rate determination by radiotelemetry in mice. AB - Mice are not commonly used in cardiovascular physiology, especially in space physiology because of methodological problems. The initial studies that have monitored arterial pressure and heart rate in mice used the tail cuff plethysmography method or classical catheterization techniques with a catheter liable to an external pressure transducer. But for long term arterial pressure measurements the studies have been facilitated by the development of radiotelemetry. This is a less constraining method as classical one allowing to monitor in continuous in freely moving animal blood pressure, heart rate and temperature. This technique allows to record these parameters thank to an implanted transmitter without physical connection with the monitoring system. The aim of this work was to valid the use of the radiotelemetry in mice to evaluate arterial blood pressure and heart rate during different stress conditions (but only control data are described in this paper). PMID- 12638627 TI - Juan J. Rorigo, MD on minimally invasive hip surgery. PMID- 12638628 TI - Research on alcoholism treatment. PMID- 12638629 TI - Assessment of alcoholic patients: advances and future challenges. AB - Noteworthy advances have been made in methods of assessing patients suffering alcohol problems. More than one hundred measures are now available to assist clinicians and researchers in screening for such problems, diagnosing them, and developing treatment plans individualized according to relevant patient characteristics. This chapter reviews progress in assessment supporting each of these activities and suggests directions for future research. It concludes by identifying a series of research issues that cut across multiple domains of alcohol assessment. PMID- 12638631 TI - Advances in research design and analysis for alcohol treatment. PMID- 12638630 TI - Biomarkers as aids to identification of relapse in alcoholic patients. AB - Since return to drinking is common in patients recovering from alcoholism, recognition of relapse should be an important component of treatment. Recurrent assessment with biochemical measures can provide clinicians with useful information on the drinking status of their patients. This chapter addresses issues surrounding the importance of early detection of relapse, describes biochemical markers that may assist in this, reviews relevant scientific investigations, and offers recommendations to researchers and clinicians. PMID- 12638632 TI - Natural resolution of alcohol-related problems. PMID- 12638633 TI - The search for a rational basis for treatment selection. PMID- 12638634 TI - Psychotherapy and motivational enhancement. PMID- 12638635 TI - Involvement of support networks in treatment. AB - Alcohol dependence and abuse are biopsychosocially determined and ameliorated. However, social factors involved in effecting treatment outcomes continue to be understudied as a context in which psychological and biological factors are researched and also as a treatment focus in their own right. Yet the client's social setting and relationships during and after treatment are more important forces in the recovery process than formally defined alcohol treatment interventions. Though this has been so historically, it is even more likely now in managed care environments where treatments are highly circumscribed. This suggests that treatment interventions refocus toward targeting changes in the patient's social environment. Treatments involving spouses and families of the patient's social networks it has been shown, are incrementally effective. A high priority for research is to identify the set of conditions under which the wider social network can also be effective. Early results are promising though not well understood. The next generation of network therapies will focus on testing the conditions under which various components of the patient's social network are important in affecting patient outcomes. As with other therapies, an important focus for research is on identifying the active ingredients in these treatments that bring about favorable change in patients generally or that enhance the outcomes of the subset of patients particularly amenable to network interventions (patient-treatment "matching"). Recommended priorities for future research in network interventions are identified. PMID- 12638636 TI - Alcoholics Anonymous and 12-step alcoholism treatment programs. AB - Alcoholics Anonymous (AA) self-help groups are the most commonly accessed component of the de facto system of care for alcohol problems in the United States. Further, AA's concepts and approach have strongly influenced a significant number of professional treatment programs. Nevertheless, only a modest number of longitudinal, comparative outcome studies on AA and on professional 12-step treatment programs have been conducted, which has limited both the certainty and scope of conclusions that can be drawn about these interventions. Research indicates that participation in Alcoholics Anonymous and in 12-step treatment are associated with significant reductions in substance abuse and psychiatric problems. Further, such interventions, it has been found, reduce health care costs over time in naturalistic, quasi-experimental, and experimental studies. Evaluation studies have also begun to illuminate the processes through which self-help groups and 12-step treatment programs exert their effects. To build on this knowledge base, future research should (1) be methodologically flexible and well-matched to its phenomenon of interest, (2) include evaluation of the unique features of self-help organizations, (3) increase representation of African-Americans and women in research samples, and (4) increase statistical power through larger sample sizes and more reliable measurement. Key content areas for future enquiry include further longitudinal evaluation of the outcomes of participation in AA and 12-step treatment (particularly in outpatient samples); better specification of the aspects of AA that influence outcome; and individual-, community-, and health organization level controlled studies of the health care cost consequences of 12-step interventions. PMID- 12638637 TI - Behavioral and cognitive-behavioral treatments for alcoholism: research opportunities. PMID- 12638638 TI - The Community Reinforcement Approach. AB - This chapter reviews two behavioral substance abuse treatments: The Community Reinforcement Approach (CRA) and Community Reinforcement and Family Training (CRAFT). Both of these programs were built on the concept that an individual's recovery is greatly affected by his or her unique environment. This environment, or reinforcing "community," is composed of family, friends, work/school, social activities, and perhaps spiritual affiliations. CRA, the first of these two programs to be developed, was created specifically for the problem drinker (Hunt & Azrin, 1973). The goal of CRA is to rearrange multiple aspects of an individual's "community" so that a clean and sober lifestyle is more rewarding than one that is dominated by alcohol and drugs. Subsequently, CRAFT was developed for the many individuals with substance abuse problems who are vehemently opposed to treatment (Institute of Medicine, 1990). CRAFT works through concerned family members and friends of these treatment refusers in an effort to get them to seek therapy (Sisson & Azrin, 1986). Descriptions and the empirical support for CRA and CRAFT follow. PMID- 12638639 TI - Acamprosate. PMID- 12638640 TI - Advances in the use of naltrexone: an integration of preclinical and clinical findings. AB - Both preclinical and clinical studies are critical in the development of effective pharmacotherapeutic approaches for treating alcoholism. Nowhere has this been more evident than in the development of naltrexone for treating alcohol relapse. As studies continue on the use of naltrexone for modifying alcohol intake, promising avenues for continued work on maximizing the efficacy of naltrexone for treating alcohol abuse and alcoholism are emerging. Recent research suggests that naltrexone can influence key components of alcohol dependence, including loss of control over the decision to drink and the amount of alcohol consumed. Although not uniformly positive, the majority of clinical trials supports the hypothesis that naltrexone can reduce the urge to drink, increase the number of days abstinent, and minimize the risk of relapse to heavy drinking. Human laboratory and preclinical paradigms that have investigated how naltrexone alters patterns of drinking suggest that naltrexone treatment results in earlier cessation of drinking within a session. In addition, preclinical data suggest that the amount of alcohol consumed declines during subsequent sessions in the presence of naltrexone. Based on this analysis, future clinical trials should consider using analytic approaches that evaluate patterns of drinking (e.g., multiple event analysis) rather than single events (e.g., survival analysis). Furthermore, behavioral interventions and instructions can also be developed to take advantage of this effect. Additional preclinical and clinical work is warranted to identify dosing strategies that ensure adequate drug levels while reducing the possibility of developing tolerance to naltrexone. Finally, studies designed to identify the characteristics of drinking populations that are responsive to naltrexone and studies investigating the potential advantage of combining naltrexone with agents that alter a number of neurotransmitter systems are exciting new avenues of research. Ultimately, these lines for research promise to provide critical information that can be used to maximize the efficacy of naltrexone for treating alcoholism. PMID- 12638642 TI - Animal models of motivation for drinking in rodents with a focus on opioid receptor neuropharmacology. AB - Ethanol, like other drugs of abuse, has motivating properties that can be developed as animal models of self-administration. A major strength of the operant approach where an animal must work to obtain ethanol is that it reduces confounds due to palatability and controls for nonspecific malaise-inducing effects. In the domain of opioid peptide systems, limited access paradigms have good predictive validity. In addition, animal models of excessive drinking-either environmentally or genetically induced-also appear sensitive to blockade or inactivation of opioid peptide receptors. Ethanol availability can be predicted by cues associated with positive reinforcement, and these models are sensitive to the administration of opioid antagonists. Perhaps most exciting are the recent results suggesting that the key element in opioid peptide systems that is important for the positive reinforcing effects of ethanol is the mu-opioid receptor. How exactly ethanol modulates mu-receptor function will be a major challenge of future research. Nevertheless, the apparently critical role of the mu receptor in ethanol reinforcement refocuses the neuropharmacology of ethanol reinforcement in the opioid peptide domain and opens a novel avenue for exploring medications for treating alcoholism. PMID- 12638641 TI - The status of serotonin-selective pharmacotherapy in the treatment of alcohol dependence. AB - Research performed during the past 20 years has shown that serotonin (5 hydroxytryptamine; 5-HT) neurotransmission is related to alcohol dependence. Both theoretical and empirical research have supported the idea that alcohol dependence is a chronic disease and that, in addition, biological vulnerabilities contribute to the pathogenesis of alcohol dependence. Preclinical studies have consistently demonstrated that there is a relationship between 5-HT function and alcohol consumption. Furthermore, there is evidence building that lends support for the existence of distinct alcoholic subtypes that may be differentiated by the type or complexity of their 5-HT dysfunction. Beyond excessive drinking, behaviors that are indicators of 5-HT dysregulation are depression, anxiety, impulsiveness, and early-onset problem drinking. This chapter will discuss the usefulness of 5-HT-selective pharmacotherapy in treating alcohol dependence and will provide both historical and current perspectives on its use. PMID- 12638643 TI - Outcomes of alcohol treatment for adolescents. AB - Alcohol and other drug use by youth continue to be an important focus for this nation. Both moderate and heavy alcohol consumption are associated with a higher risk of alcohol-related medical consequences and accidental injuries for youth. Despite knowledge of possible consequences, a high percentage (30%) of high school students nationwide reports episodes of hazardous drinking (five or more drinks on one occasion). Increased awareness and concern related to adolescent substance use has led to the outgrowth of additional treatment facilities and programs for this age group. This chapter examines the impact of developmental factors on the assessment process and subsequent treatment of adolescent alcohol use disorders. In addition, treatment outcome research, intervention studies, relapse, and factors that may influence the recovery process of youth are discussed. PMID- 12638644 TI - Treatment for alcohol-related problems: special populations: research opportunities. AB - For the subgroups indicated, a few questions/issues are relevant to all three (women, elderly, minorities): 1. Heterogeneity of the special populations, for example, Hispanic-Americans are from different countries with different cultures. Women and the elderly vary by age, education, income, social class, health status, etc., to say nothing of ethnicity/color/religion. 2. Of therapy modalities, professional and indigenous, which are more efficacious? 3. Are group specific therapies needed, or will sensitivity to a particular group work as well? WOMEN: Stereotypes and myths have prevailed, for example, the long-standing belief that women have poorer prognoses than male alcoholics. When female and male alcoholics are compared, women report more positive family history, a later onset of drinking and problems, more marital disruption, more comorbidity, etc. The review of treatment outcomes (Vannicelli, 1986) showed few significant gender differences in outcomes. Research recommendations include biological and genetic studies, women's view of and use of therapeutic modalities, and outcome studies of different modalities, including all female facilities. ELDERLY: Medications are used more by older patients, and such patients are more likely to experience adverse drug reactions. In the moderate social use of alcohol, there are conflicting reports and the extent of elderly use awaits decisive study. The etiology of problem drinking by older persons is studied rarely. An attempt has been made to explain onset later in life (vs. earlier onset) based on the stresses of aging (loss, loneliness, health problems, etc.); research results have not been supportive. Consequences of older persons' heavy drinking seems to be most often alcohol-related medical disorders, although there are often familial and social consequences. Atkinson (1995) recommended the development of elder-specific outcome measures, study of the efficacy of different treatment modalities, and study of the efficacy of treatment for patients in elder-specific and mixed age groups, etc. MINORITIES: Each of the federally mandated minority groups in the United States is heterogeneous. The epidemiology of use and abuse of alcohol and other drugs is well studied, but treatment issues are not. AMERICAN INDIANS: There are more than 200 tribes; each has its own customs and culture. Some tribes are abstinent; others have big problems with abuse of alcohol, and other drugs. Orthodox treatment methods, used by professional counselors and therapists, have not worked very well. RECOMMENDATION: study of traditional Indian forms of healing practices combined with other treatment; this would be a culture-sensitive model. BLACK AMERICANS: This includes not only African-Americans but people from the Caribbean, Central and South America, etc. Among African-Americans, there is a history of ambivalence toward alcohol: on the one hand, a tolerant "nightclub culture" and on the other, church beliefs in temperance and abstinence. There is "respectable drinking" and "problem drinking," most often defined as solitary or public drinking. The primary source of support is considered familial, so people tend to be distrustful of therapy from "strangers." They are anonymous in promoting sobriety and study of subcultural norms and the history of slavery. Earlier ethnographic works (Liebow, 1967) were of "street-corner men," slum dwellers, ghetto norms; recommended: studies of middle-class African-American life and drinking behaviors. ASIAN AMERICANS: A study in Los Angeles reports differences among Chinese, Japanese, Filipinos, and Koreans in drinking beliefs and behavior. Of these groups, the Japanese in Japan and the Japanese-Americans report the largest number of heavy drinkers. It is, however, considered a private matter, even when associated with social problems. Interestingly, there is an organization called the All Nippon Sobriety Association (like Alcoholics Anonymous). RECOMMENDATIONs: studies of generational differences among Japanese-Americans in use and efficacy of treatment. For the Chinese-Americans, who are fairly permissive about older persons' drinking and share a belief in the health benefits of alcohol, a gender/gerontological study is recommended. HISPANIC-AMERICANS: As a total group, they drink more and present more alcohol-related problems than other immigrant minorities. Age, ethnicity, and gender patterns in permissiveness to drink need to be explored. Treatment sought is often in pentecostal churches and Centros for Espiritismo. Hispanics are not likely to seek help in formal clinical settings which emphasize alcohol consumption as the basic, core problem. They are more likely to seek out and be responsive to the perception of their drinking problem as sin and a rejection of Jesus. It is not that minorities do not recognize problems and seek out help. They are not likely to seek out the health profession's offering of outpatient clinics, residential treatments, etc. PMID- 12638645 TI - Treatment of tobacco dependence in alcoholics. AB - Because of the high morbidity and mortality that alcoholic smokers experience from tobacco-caused diseases, treatment for tobacco dependence among alcoholics is warranted. Much progress has been made during the last decade in addressing tobacco dependence in alcoholism treatment units. Treatment of tobacco dependence in alcoholic smokers does not seem to cause excessive relapse to drinking and, in fact, stopping smoking may enhance abstinence from drinking. Therefore, treatment for alcoholic smokers should take place whenever and wherever the patient comes in contact with the health care system. Because alcoholic smokers as a rule are more dependent on nicotine than their nonalcoholic counterparts, they may need more intensive pharmacotherapy and behavioral therapy. Because many of them have experienced 12-step approaches to recovery, that same technology can be used to initiate and maintain abstinence from tobacco use. Moreover, several pharmacologic options exist to treat tobacco dependence in alcoholic smokers. However, the efficacy of several pharmacologic therapies for alcoholic smokers needs to be tested. In addition, further research is needed on effective treatments for recovering alcoholics of various racial/ethnic backgrounds. PMID- 12638646 TI - Alcohol and psychiatric comorbidity. AB - Comorbid psychiatric disorders and drug use disorders (DUDs) are common among alcoholics (Regier, Farmer, Rae, Locke, Keith, Judd, & Goodwin, 1990; Kessler, McGonagle, Zhao, Nelson, Hughes, Eshleman, Wittchen, & Kendler, 1994). These comorbid disorders often predict a shorter time to relapse of alcoholism (Greenfield, Weiss, Muenz, Vagge, Kelly, Bello, & Michael, 1998). However, despite the prevalence and the adverse effects of this comorbidity, few controlled treatment studies have been conducted involving this dual diagnosis population (Litten & Allen, 1999). To date, most of these few studies of alcoholics with comorbid disorders have been restricted to studies of alcoholics with either comorbid major depression or comorbid anxiety disorders (Litten & Allen, 1995). The results of these trials suggest efficacy for SSRI antidepressants and tricyclic antidepressants for treating alcoholics with comorbid major depression and suggest efficacy for buspirone for treating alcoholics with comorbid anxiety disorders (Mason, Kocsis, Ritvo, & Cutler, 1996; Cornelius, Salloum, Ehler, Jarrett, Cornelius, Perel, Thase, & Black, 1997; Kranzler, Burleson, Del Boca, Babor, Korner, Brown, & Bohn, 1994). However, controlled treatment studies involving alcoholics with other comorbid disorders are almost totally lacking. Consequently, to date, no empirically proven treatment exists for most of these comorbid disorders. PMID- 12638647 TI - Brief interventions and the treatment of alcohol use disorders: current evidence. PMID- 12638648 TI - Spirituality, treatment, and recovery. PMID- 12638649 TI - Research priorities for alcoholism treatment. AB - The goals of the current NIAAA review process were to assess the breath, coverage, and balance of the treatment research portfolio and to identify areas for increased attention. Both alcoholism treatment experts and non-alcohol related treatment experts were called upon to help with this task. Our group was encouraged by the fact that results from previous reviews have effectively been incorporated into the institute portfolio, particularly in the areas of the pharmacotherapy and psychotherapy development programs. The present review identified the five priority areas: mechanisms of action of treatment, combination and sequencing of treatments, health-seeking patterns and processes, concurrent disorders, and understanding help agent behaviors. In addition, several methodological and/or conceptual issues emerged that it was felt, crosscut these areas. These issues included greater emphasis on intervention testing in special populations, patient-treatment matching, measurement techniques, and technology transfer. Finally, several mechanisms of funding were outlined, including across-institute initiatives that could facilitate larger scale translational work such as comorbidity studies and/or studies of combined treatments. All the above were deemed important by the advisory committee, but an overarching emphasis was placed on understanding the mechanisms of action of treatment. For example, it is important to know that a particular pharmacotherapy reduces craving and also how this comes about. Similarly, it is key to the continued refinement of psychosocial treatments that the active components and their mechanisms of change be clarified. Such mechanism studies are crucial for the development of more heuristic and more accurate theoretical formulations that must guide our understanding of alcohol dependence if treatment, relapse, and recovery are to become more scientifically grounded. PMID- 12638650 TI - Induction of glutathione-S-transferase in the Northern quahog Mercenaria mercenaria after exposure to the polychlorinated biphenyl (PCB) mixture Aroclor 1248. AB - The glutathione-S-transferases (GSTs) from the Northern quahog (Mercenaria mercenaria) were examined after an injection of a polychlorinated biphenyl (PCB) mixture, Aroclor 1248, to a concentration of approximately 50 ppm. Enzymatic analysis indicated a fourfold increase in the GST activity of quahogs injected with PCBs compared with that of the control. An electrophoretic analysis of the GST from the PCB-exposed quahogs showed a 1.5-fold increase in the concentration over that of the control. Purification of the GST on a glutathione affinity column yielded a glutathione binding protein, in addition to the GSTs. However, the amount of the glutathione binding protein in the PCB-injected quahogs was found to decrease by approximately 50% in comparison to the glutathione binding protein in the control quahogs. PMID- 12638651 TI - Purification and characterization of a phosphodiesterase from Bothrops alternatus snake venom. AB - A phosphodiesterase was purified from the venom of the snake Bothrops alternatus by a combination of gel filtration and ion exchange chromatographies. In SDS PAGE, the enzyme gave a single band with a molecular mass of 105 kDa, which was unaltered in the presence of beta-mercaptoethanol, indicating that the protein contained no subunits. A single protein band was also observed in native PAGE. There were no contaminating 5'-nucleotidase, alkaline phosphatase and protease activities. The enzyme was recognized by commercial bothropic antiserum and gave a single band in immunoblotting. The enzyme had a pH optimum in the range of 7.5 9.5 and the optimum temperature was 60 degrees C, with activity being rapidly lost within 1 min at > or = 70 degrees C. The Km of the enzyme was 2.69 mM. PDE activity was potentiated by cobalt and, to a lesser extent, by calcium, whereas copper, manganese, zinc, EDTA, and beta-mercaptoethanol were inhibitory. These properties show that this enzyme is very similar to that isolated from other snake venoms. PMID- 12638652 TI - Use of amphotericin B as optical probe to study conformational changes and thermodynamic stability in human serum albumin. AB - The binding of polyene antibiotic amphotericin B to serum albumin was studied using absorption, fluorescence, and circular dichroism techniques. A hypochromic effect was observed in the absorption spectrum of amphotericin B in the presence of albumin with maxima at 366 nm, 385 nm, and 408 nm, which correspond to the absorption of the monomeric form of amphotericin B. A modification on the circular dichroism spectrum of amphotericin B in the presence of albumin was observed at bands 329 nm and 351 nm (excitronic interaction), which suggests that only amphotericin B monomer is bound to the protein. Amphotericin B perturbs the specific markers for sites I, II, and fatty acid binding site bound to these sites, suggesting that amphotericin B interacts with a great binding area in albumin. Lysines 199 and 525 in albumin participate in the molecular interaction between amphotericin B and the protein. The absorption spectrum of amphotericin B bound to albumin was sensitive to the chemical and thermal treatment of the protein, to neutral-basic transition of albumin and to conformational changes induced by the binding of other ligands to this protein. PMID- 12638654 TI - Effects of different buffers on the thermostability and autolysis of a cold adapted protease MCP-01. AB - A cold-adapted protease MCP-01 was obtained from deep-sea psychrotrophic bacterium Pseudoaltermonas sp. SM9913. The effects of four different buffers, all at 50 mmol/l concentration, on its thermostability and autolysis were studied. The autolysis process of MCP-01 was studied by capillary electrophoresis. The thermostability of MCP-01 increased successively in the following order: carbonate < Tris < phosphate < borate. The optimum temperature for casein hydrolysis also increased in the same order. This suggested that the conformation of MCP-01 was flexible and its autolytic susceptibility was affected by some factors in the buffers such as charge and ionic species. The results also showed that different buffers, in addition to affecting the autolysis speed, gave different patterns of autolysis products. In carbonate buffer, Tris buffer, phosphate buffer and borate buffer, the autolysis patterns of MCP-01 were different. These results suggested that protease MCP-01 probably have different conformations in different buffers, thus exposing different autolysis sites on the enzyme surface. In addition, the loss of activity correlated with the speed of autolysis in the four different buffers, showing that autolysis may be a reason for the low thermostability of the enzyme. PMID- 12638653 TI - The mitochondrial tricarboxylate carrier of silver eel: dimeric structure and cytosolic exposure of both N- and C-termini. AB - The mitochondrial tricarboxylate carrier plays a fundamental role in the hepatic fatty acid synthesis. In this study, we investigated the transmembrane organization of this protein in the inner membrane of eel liver mitochondria using anti-N-terminal and anti-C-terminal antibodies. These antibodies recognized the N- and C-termini of the tricarboxylate carrier in intact mitoplasts, thus suggesting a cytosolic exposure of these regions in the membrane-bound protein. This structural arrangement of the tricarboxylate carrier was further confirmed by protease treatment of intact mitoplasts. Moreover, the oligomeric state of the native tricarboxylate carrier was investigated by blue native electrophoresis. A dimeric form of the carrier protein was found when eel liver mitochondria were solubilized with the mild detergent digitonin. These findings suggest an arrangement of the dimeric tricarboxylate carrier into an even number of membrane spanning domains, with the N-terminal and C-terminal regions oriented toward the intermembrane space of fish mitochondria. PMID- 12638655 TI - Metallohistins: a new class of plant metal-binding proteins. AB - Two small multimeric histidine-rich proteins, AgNt84 and Ag164, encoded by two nodule-specific cDNAs isolated from nodule cDNA libraries of the actinorhizal host plant Alnus glutinosa, represent a new class of plant metal binding proteins. This paper reports the characterization of the purified in vitro expressed proteins by size exclusion chromatography, circular dichroism, equilibrium dialysis, metal affinity chromatography coupled with mass spectrometry, and nuclear magnetic resonance spectroscopy. These analyses reveal that each polypeptide is capable of binding multiple atoms of Zn2+, Ni2-, Co2+, Cu2+, Cd2+ and Hg2+. A reversible shift in histidine Cepsilon1 and Cdelta2 protons in NMR analysis occurred during titration of this protein with ZnCl2 strongly suggesting that histidine residues are responsible for metal binding. AgNt84 and Ag164 are not related to metal binding metallothioneins and phytochelatins and represent a new class of plant metal binding proteins that we propose to call metallohistins. Possible biological roles in symbioses for AgNt84 and Ag164, and their potential for use in bioremediation are discussed. PMID- 12638656 TI - Construction of hypothetical three-dimensional structure of P2Y1 receptor based on Fourier transform analysis. AB - G protein-coupled receptors constitute a large family of homologous transmembrane proteins that represents one of the most important classes of confirmed drug targets. For novel drug discovery, the 3D structure of target protein is indispensable. To construct hypothetical 3D structures of G protein-coupled receptors, several prediction methods have been proposed. But none of the them has confirmed a correct ligand binding site. In this study we constructed the 3D structure of bovine rhodopsin using the prediction method proposed by Donnelly et al., with some modification. We found that our 3D model showed a good agreement with the reported retinal binding site. Using the similar method, we constructed the 3D structure of the P2Y1 receptor; one of the G protein-coupled receptors, and showed a binding site of an endogenous ligand, ADP, on the basis of the 3D model and in vitro experimental data. These results should be valuable for design of a specific antagonist for P2Y1 receptor. PMID- 12638657 TI - Time-resolved phosphorescence of tyrosine, tyrosine analogs, and tyrosyl residues in oxytocin and small peptides. AB - We present the time-resolved phosphorescence of oxytocin, two oxytocin derivatives, vasopressin and a series of compounds that serve as models for free tyrosine. One of the oxytocin derivatives, desaminodicarbaoxytocin, has the disulfide bridge replaced by an ethylene bridge, and lacks the N-terminus. Similar to the reported fluorescence decays of tyrosine in these peptides, the phosphorescence decays generally are not single exponentials, but can be fit as biexponentials. The decay times for the oxytocin peptides are shorter than for desaminodicarbaoxytocin or the model compounds, and this we attribute to enhanced spin-orbit coupling due to the presence of sulfur. We measured the phosphorescence decay of the model cyclic pentapeptide that contains tyrosine and compared it to that observed for the same cyclic pentapeptide in which tyrosine is replaced by tryptophan. We also report the phosphorescence of 2-tryptophan oxytocin, and deamino-2-tryptophan-oxytocin in which biexponential phosphorescence decay is also observed. PMID- 12638658 TI - Heterogeneity of caprine beta-casein elucidated by RP-HPLC/MS: genetic variants and phosphorylations. AB - Casein variants occurring in milks from goats homozygous at the alpha(s1)-Cn locus were separated and identified by an RP-HPLC/ESI-MS method. Preferential haplotypes arose as well as some particularities in posttranslational modifications. A new variant of caprine beta-Cn, named C, as well as the phosphorylations pattern of the protein were characterized by the combined use of peptide mass fingerprinting and sequencing by tandem mass spectrometry. The molecular mass of the new variant in its 6P form was measured as 23854 Da and it differs in a mono amino acid substitution, A177 --> V177, from the variant A. The phosphorylation pattern of caprine beta-Cn is homologous to bovine beta-Cn concerning the 5P located on Ser15, 17, 18, 19, 35 but it presents a specific additional phosphorylation site on Thr12 that is comparable to human beta-Cn phosphorylation located on Thr3. PMID- 12638659 TI - Direct extraction of DNA from soils for studies in microbial ecology. AB - Molecular analyses for the study of soil microbial communities often depend on the extraction of DNA directly from soils. These extractions are by no means trivial, being complicated by humic substances that are inhibitory to PCR and restriction enzymes or being too highly colored for blot hybridization protocols. Many different published protocols exist, but none have been found to be suitable enough to be generally accepted as a standard. Most direct extraction protocols start with relatively harsh cell breakage steps such as bead-beating and freeze thaw cycles, followed by the addition of detergents and high salt buffers and/or enzymic digestion with lysozyme and proteases. After typical organic extraction and alcohol precipitation, further purification is usually needed to remove inhibitory substances from the extract. The purification steps include size exclusion chromatography, ion-exchange chromatography, silica gel spin columns, and cesium chloride gradients, among others. A direct DNA extraction protocol is described that has been shown to be effective in a wide variety of soil types. This protocol is experimentally compared to several published protocols. PMID- 12638660 TI - Using flow cytometry to quantify microbial heterogeneity. AB - Flow cytometry is a powerful technique for the study of single cells, and thus it is of particular utility in the study of heterogeneity in microbial populations. This review seeks to highlight the role of flow cytometric analyses in studies of microbial heterogeneity, drawing wherever possible on recently published research articles. Whilst microbial heterogeneity is well documented in both natural and laboratory environments, the underlying causes are less well understood. Possible sources for the heterogeneity that is observed in microbial systems are discussed, together with the flow cytometric tools that aid its study. The role of flow cytometry in molecular biology is discussed with reference to gene reporter systems, which enable heterogeneity of gene expression to be monitored. With the recent sequencing of a variety of microbial genomes, it is anticipated that flow cytometry will have an increasing role to play in studying the effects of gene expression and mutation on heterogeneity, and in resolving the interactions of genetics and physiology. PMID- 12638661 TI - Genomics and bacterial metabolism. AB - The field of bacterial metabolism and physiology is arguably the oldest in microbiology. Much of our understanding of biological processes and molecular paradigms has its roots In early studies of prokaryotic physiology. After a period of declining interest in metabolic studies (prompted by the insurgence of molecular techniques), genomic technologies are revitalizing the study of bacterial metabolism and physiology. These new technologies bring a means to approach metabolic questions with a global perspective. When used in combination with classical and molecular techniques, emerging global technologies will make it feasible to understand the complex integration of metabolic processes that result in an efficient physiology. At the same time, without increased computational capabilities, the massive amounts of data generated by these technologies threaten to overwhelm, rather than facilitate, this work. For genomic technologies to reach their potential for increasing our understanding of bacterial metabolism, microbiologists must become more collaborative and multidisciplinary than at any time in our history. PMID- 12638662 TI - The President's physician. PMID- 12638664 TI - Janet G. Travell, MD: a daughter's recollection. PMID- 12638663 TI - Cardiology and myofascial trigger points: Janet G. Travell's contribution. PMID- 12638665 TI - Off-pump coronary artery bypass grafting and transmyocardial laser revascularization via a left thoracotomy. AB - Off-pump coronary artery bypass grafting may be combined with adjunctive transmyocardial laser revascularization to optimize revascularization. This approach may be advantageous for high-risk patients, particularly those having undergone previous sternotomies. From October 2000 through May 2001, 17 patients (9 women and 8 men) underwent off-pump coronary artery bypass grafting and transmyocardial laser revascularization via a left thoracotomy. The patients had a mean age of 63 years and a mean ejection fraction of 0.33. All but 1 patient had undergone previous coronary surgery. In each patient, the heart was approached via a left thoracotomy through the 5th intercostal space, and 37 transmural channels, 1 mm in diameter, were each created with a single pulse of the carbon dioxide laser. Coronary artery bypass grafting was then performed with left internal thoracic artery or saphenous vein grafts. The follow-up period ranged from 2.1 to 9.3 months (mean, 6.2 months). The patients received 28 bypass grafts (mean, 1.6 grafts). Postoperatively, 2 patients required inotropic support. On day 8, 1 patient died of ventricular fibrillation. After a mean hospitalization of 7.7 days, the remaining patients were discharged, free of angina. At follow-up examination after a mean of 6 months (range, 2-9 months), 15 patients remained free of angina and one had mild angina. None had required further hospitalization. Performed via a left thoracotomy, off-pump coronary artery bypass grafting plus transmyocardial laser revascularization yielded an acceptable mortality rate, no major morbidity, and substantial angina relief in this carefully selected group of challenging, high-risk patients. PMID- 12638666 TI - Comparison of left ventricular electromechanical mapping and left ventricular angiography: defining practical standards for analysis of NOGA maps. AB - We performed this prospective cohort study to correlate the findings of left ventricular angiography (LVA) and NOGA left ventricular electromechanical mapping (LVEM) in the evaluation of cardiac wall motion and also to establish standards for wall motion assessment by LVEM. Fifty-five patients (35 men; mean age, 60.4 +/- 11.8 years) eligible for elective left cardiac catheterization underwent LVA and LVEM. Wall motion scores, LV ejection fractions (LVEF), and LV volumes derived from LVA versus LVEM data were compared and analyzed statistically. Receiver operating characteristic (ROC) curves were used to assess the accuracy of LVEM in distinguishing between normal, hypokinetic, and akinetic/dyskinetic wall motion. Mean LVEM procedure time was 37 +/- 11 minutes. The LVEM and LVA findings differed for mean LVEF (55% +/- 13% vs 36% +/- 9%), mean end-systolic volume (56 +/- 13 mL vs 36 +/- 10 mL), and mean end-diastolic volume (174 +/- 104 mL vs 123 +/- 65 mL). Mean wall motion scores (+/- SD) for normokinetic, hypokinetic, and akinetic/dyskinetic segments were 13.9% +/- 5.6%, 8.3% +/- 5.2%, and 3.2% +/- 3.1%, respectively. Cutpoints for differentiating between wall motion types were 12% and 6%. The ROC curves showed LVEM to have a 93% accuracy in differentiating between normokinetic and akinetic/dyskinetic segments and a 73% accuracy between normokinetic and hypokinetic segments. These data suggest that LVEM can differentiate between normal and abnormal cardiac wall motion, although it is more accurate at differentiating between normokinetic and akinetic/dyskinetic motion than between normokinetic and hypokinetic motion. PMID- 12638667 TI - Recent surgical experience in chronic constrictive pericarditis. AB - We reviewed the cases of 42 consecutive patients who underwent surgery for chronic constrictive pericarditis between January 1994 and January 2002, to determine causes, surgical techniques, mortality and morbidity rates, and improvement of functional capacity. Thirty-one of our patients were men (73.8%) and 11 were women (26.2%); their average age was 39.3 years. The pericarditis was tuberculous in 24 patients (57.1%), idiopathic in 10 (23.8%), rheumatic in 4 (9.5%), uremic in 2 (4.8%), and neoplastic in 2 (4.8%). In all cases, our approach was via median sternotomy. All anterior pericardium was resected from phrenic nerve to phrenic nerve. In 4 of 18 patients with calcific pericardial plaques, we could not develop a cleavage plane, so we left the plaques in place, reducing their size and relieving constriction by means of wedge incisions that reached the epicardium. No patient required cardiopulmonary bypass due to severe calcification or adhesion, but we did perform bypass with no difficulty in 5 patients who required additional cardiac surgery The early postoperative mortality rate was 11.9% (5 patients). In the 1st postoperative month, functional capacity improved dramatically: the number of patients in New York Heart Association functional class IV moved from 18 (preoperatively) to 1; in class III, from 20 to 6; in class II, from 4 to 13; and in class I, from 0 to 22. We recommend the median sternotomy approach for chronic constrictive pericarditis and consider cardiopulmonary bypass safe to use in indicated cases. PMID- 12638668 TI - Comparison of intraoperative transit-time flow measurement with early postoperative magnetic resonance flow mapping in off-pump coronary artery surgery. AB - The purpose of this prospective study was to evaluate graft patency of off-pump coronary artery surgery intraoperatively by transit-time flow measurement and to compare this technique with postoperative magnetic resonance flow mapping. Twenty patients (13 men and 7 women; mean age, 679 +/- 76 yr) underwent off-pump coronary artery surgery. Intraoperative transit-time flow measurement of grafts was performed measuring maximum, minimum, and mean flows. For each graft, the pulsatile index was calculated by dividing the difference between the maximum and the minimum flow by the mean flow In the early postoperative period (1st week), magnetic resonance flow mapping was performed using phase contrast flow quantification. Mean intraoperative flow values and mean magnetic resonance flow mapping values were compared. At the same postoperative session, contrast enhanced magnetic resonance angiography was performed to evaluate graft patency. In 20 patients, a total of 49 coronary graft flows were assessed with intraoperative transit-time flow measurement and postoperative magnetic resonance flow mapping. Upon comparison, there was a strong correlation between techniques, with stable and statistically significant differences between the intraoperative and postoperative flow mapping values. One saphenous vein graft was revised intraoperatively, due to graft failure. Our data suggest that the combined use of intraoperative transit-time flow measurement and postoperative magnetic resonance flow analysis has a potential role in the assessment of graft patency in off-pump coronary artery surgery although more study is required. PMID- 12638669 TI - Tricuspid valve detachment in closure of congenital ventricular septal defect. AB - From January 1991 through December 2001, 600 patients underwent closure of a perimembranous ventricular septal defect through a right atrial approach at our institution. In 122 of these patients, the operation included temporary detachment of a tricuspid valve septal leaflet from the annulus to allow complete visualization of a perimembranous ventricular septal defect The mean age of the patients at surgery was 4.6 years in those who underwent leaflet detachment and 4.7 years in the 478 patients who did not (P > 0.05). Preoperatively, all patients were in sinus rhythm. Echocardiography showed trivial tricuspid regurgitation in 21 of the patients undergoing detachment and in 39 of the non detachment patients. There was no difference in bypass time or aortic cross-clamp time between the 2 groups. Postoperatively, 3 patients in the non-detachment group had heart block; all other patients were in sinus rhythm. Echocardiograms on the 7th postoperative day showed small residual ventricular septal defects in none of the patients who underwent valve detachment and in 10 of the non detachment patients; mild tricuspid regurgitation was present in 12 non detachment patients only; and trivial tricuspid regurgitation was present in 19 patients who underwent valve detachment and in 29 who did not. There was no hospital death in either group. Long-term follow-up showed no progression of tricuspid regurgitation or tricuspid stenosis. All patients remained in sinus rhythm. This study suggests that tricuspid valve detachment is a safe, effective technique that improves exposure for ventricular septal defect repair and does not adversely affect valve competence. PMID- 12638670 TI - Plantaris tendon graft for atrioventricular valve repair: a novel hypothetical technique. AB - Heart valve repair is a biomaterial-dependent procedure, yet autogenous tissue for valvular reconstruction has to date been obtained mostly from the pericardium and fascia lata. Most clinical studies recommend valve repair as an alternative to replacement. We now put forward a hypothesis, extrapolated from hand surgery, for use of the plantaris tendon in heart valve repair. This proposal, if implemented, would increase the supply of autogenous donor tissue for valve repair, thereby enhancing the surgeon's armamentarium. The report describes a novel technique that in our judgment warrants future clinical development. PMID- 12638671 TI - Inappropriate shock delivery and biventricular pacing cardiac defibrillators. AB - In the United States, physicians adapt currently available defibrillators to accommodate leads for biventricular pacing in those congestive heart failure patients who might benefit from cardiac resynchronization and who are additionally at risk for sudden cardiac death. The adaptation of the lead system of available defibrillators to also allow them to function as biventricular pacemakers presents occasions in which inappropriate shocks are delivered due to double counting of the right and left ventricular depolarizations by the implantable cardiac defibrillator. We reviewed a series of inappropriate shock deliveries that occurred after the implantation of biventricular pacing cardiac defibrillators at our institution; all of these shocks were related to ventricular double counting. Each had different underlying causes and management strategies. Complications such as these emphasize the importance of attentiveness to ventricular channel electrograms and to device sensing with the use of biventricular pacing cardiac defibrillators. In addition, a thorough working knowledge of pacemaker and defibrillator operation is essential for the prediction and correction of inappropriate therapies. PMID- 12638672 TI - Support with the BVS 5000 assist device during treatment of acute giant-cell myocarditis. AB - Giant-cell myocarditis is a rare and aggressive form of myocarditis with a high mortality rate. Our purpose is to summarize 3 cases of acute giant-cell myocarditis that illustrate possible outcomes with mechanical support. We reviewed the cases of 3 patients, aged 39 to 59 years, who had giant-cell myocarditis (confirmed by myocardial biopsy). The indication for ventricular assist was circulatory failure despite maximal medical treatment with 2 or more inotropic agents and intraaortic balloon pump support. Immunosuppression and a biventricular mechanical assist (BVS 5000) were used to treat all these patients. The mean duration of mechanical support was 15.7 days (range, 10 to 19 days). One patient had recovery of myocardial function and was weaned from mechanical support. This case is, to our knowledge, the first reported of ventricular support leading to cardiac recovery after diagnosis of giant-cell myocarditis. The 2nd patient was not a candidate for further surgery and died of multisystem organ failure. The 3rd patient underwent orthotopic heart transplantation after 18 days of support and was discharged. We conclude that patients with giant-cell myocarditis tend to have biventricular involvement and can recover myocardial function on mechanical support and immunosuppression. If recovery is not observed, transplantation is warranted. By avoiding left ventricular cannulation, the BVS 5000 is well suited for bridging to recovery, transplantation, or long term support. PMID- 12638673 TI - Heparin-induced thrombocytopenia with associated thrombosis in children after the Fontan operation: report of two cases. AB - Heparin-induced thrombocytopenia is a widely recognized clinical disorder. The spectrum of disease ranges from clinically insignificant to severe thrombosis (heparin-induced thrombocytopenia with associated thrombosis). Overall, thrombosis occurs in approximately 33% of adults diagnosed with heparin-induced thrombocytopenia and has been associated with high morbidity and mortality rates. Diagnostic testing for this disorder is not standard in children with thrombocytopenia who are receiving heparin, despite the fact that children with congenital heart disease may be exposed to heparin frequently. There are few reported cases of heparin-induced thrombocytopenia with associated thrombosis in children; herein, we describe the cases of 2 children who developed this disorder after undergoing a Fontan operation. PMID- 12638674 TI - Occult cardiac lymphoma presenting with cardiac tamponade. AB - Subxiphoid pericardiostomy is the procedure of choice for treatment of a pericardial effusion with tamponade. We report a case in which this procedure not only failed to reveal the presence of an occult malignancy, but also resulted in a recurrent symptomatic effusion. PMID- 12638675 TI - Early stabilization of traumatic aortic transection and mitral valve regurgitation with extracorporeal membrane oxygenation. AB - We report a case of life-threatening aortic transection with concomitant mitral papillary muscle rupture and severe lung contusion caused by a failed parachute jump. This blunt thoracic injury was treated by early stabilization with extracorporeal membrane oxygenation followed by successful delayed graft repair of the descending aorta and mitral valve replacement with a mechanical prosthesis. PMID- 12638676 TI - Atrial septal aneurysm with right-to-left interatrial shunting. AB - Interatrial shunting in the presence of an atrial septal aneurysm is an uncommon but well recognized abnormality. Previous case reports have demonstrated that elevated right atrial pressure secondary to pulmonary embolism or right ventricular infarction may cause right-to-left interatrial shunting in the presence of an atrial septal aneurysm. We describe a unique situation in which an atrial septal aneurysm was associated with a right-to-left shunt secondary to severe systemic hypotension and normal right atrial pressure. In this patient, we used midodrine, an oral alpha-1 agonist, to increase systemic arterial pressure, decrease the severity of the shunt, and treat the severe hypoxemia. This case establishes that right-to-left interatrial shunting can result from a decrease in left ventricular afterload with normal right atrial pressure. Oral alpha-1 agonist therapy can be used successfully to treat patients such as ours and possibly others with similar functional abnormalities. PMID- 12638677 TI - Right ventricular bronchogenic cyst. AB - We report an exceedingly rare case of primary bronchogenic cyst in the outflow tract of the right ventricle in a 48-year-old woman. In our review of the world literature, we found only 1 other report of an intracardiac bronchogenic cyst. Our patient's only symptom was mild dyspnea not associated with physical exertion, and the cyst was resected successfully. We report clinical aspects of the case, diagnostic methods, surgical management, and histopathologic findings. PMID- 12638678 TI - Large B-cell lymphoma of the atria. AB - We report a case of a large, B-cell lymphoma of the atria in a 65-year-old man who presented with obstructive right-heart failure, shortness of breath, cirrhosis, and ascites. A computed tomographic scan revealed a large cardiac tumor occupying both atria. The patient underwent debulking of the tumor and postoperative chemotherapy. Six months postoperatively he was alive and his symptoms of obstructive right-heart failure had improved; however, he had developed brain metastasis. PMID- 12638679 TI - Drug-induced toxic myocarditis. AB - A 66-year-old woman presented with new-onset complete left bundle branch block and congestive heart failure. She had had chronic paranoid schizophrenia for 35 years and had been taking medications to control her psychiatric disorder for the past 10 years. A 2-dimensional echocardiogram performed at the onset of congestive heart failure showed dilated cardiomyopathy with global impairment of left ventricular function (ejection fraction <0.25). Despite withdrawal of the medications most likely responsible for the heart problems (perphenazine, 2 mg; and amitriptyline, 25 mg), the patient died of refractory congestive heart failure 2 years later. Histologic examination at autopsy showed evidence of persistent toxic myocarditis with fibrosis of the heart and persistent chronic hepatitis. These autopsy findings were considered to be drug related. PMID- 12638680 TI - Familial cardiac myxoma: Carney's complex. AB - We report a case of Carney's complex in a 12-year-old boy who had the characteristic features of multiple cutaneous tumors, pigmentation, and biatrial myxoma. His large right atrial myxoma almost occluded the tricuspid valve and presented a life-threatening emergency. Surgery saved his life, but recurrence of myxoma was noted on follow-up. The familial nature of the condition is highlighted by the case of the patient's 44-year-old mother, who also presented with features of Carney's complex: multiple cutaneous tumors and a tiny, asymptomatic, left atrial myxoma, which was detected during routine echocardiographic screening. PMID- 12638681 TI - Pacemaker migration into the pouch of Douglas. PMID- 12638682 TI - Pacemaker lead entrapment syndrome. PMID- 12638683 TI - QT interval lengthening after fasting complicated by a sudden attack of torsades de pointes. PMID- 12638684 TI - Prevention of embolization of a displaced atrial septal occluder. PMID- 12638686 TI - Beyond the suprachiasmatic nucleus. AB - The suprachiasmatic nucleus is the master oscillator controlling circadian rhythms in mammals. Yet extensive temporal restructuring of behavior can occur without participation of the suprachiasmatic nucleus. This raises questions about current thinking about how to cope with jet lag and shift work. PMID- 12638685 TI - Who assisted whom? PMID- 12638687 TI - Effect of cage enrichment on the daily use of running wheels by Syrian hamsters. AB - Institutional animal care committees may one day require for the welfare of captive hamsters more floor space and the introduction of tunnels and toys. As hamsters are popular animal subjects in chronobiological research, and as clock phase is usually measured through running wheel activity, it is important to determine what effect cage enrichment might have on daily wheel use. Here the daily number of wheel revolutions, the daily duration of the running activity phase, the phase relationship between lights-off and onset of running activity, and the free-running period of circadian activity rhythms were measured in Syrian hamsters, Mesocricetus auratus, housed in single cages or in multiple cages linked by tunnels and supplied with commercial wooden toys. Free-running periodicity was not affected by cage enrichment. In multiple-cage systems, there were fewer daily revolutions, shorter wheel-running activity phases, and delayed running activity onsets. These effects, however, were small as compared to interindividual and week-to-week variation. They were statistically significant only under a light:dark cycle, not in constant darkness, and only when interindividual variation was eliminated through a paired design or when the number of cages was increased to five (the maximum tested). Daily wheel use is thus affected by cage enrichment, but only slightly. PMID- 12638688 TI - Effect of light during lactation on the phasic and tonic responses of the rat pacemaker. AB - The circadian system in mammals generates endogenous circadian rhythms and entrains them to external cycles. Here, we examine whether the lighting conditions under which rats are reared affect the properties of the circadian pacemaker. We maintained three groups of rats under constant darkness (DD-rats), constant bright light (LL-rats) or light-dark cycles of 24 hours (LD-rats) during lactation. We then studied motor activity rhythm under constant light of four intensities, and under seven light-dark cycles with periods ranging between 22 and 27 hours. Results show that neither the tau nor the phase angle to the external cycle differed between groups. Differences were found in the amplitude of the circadian rhythm and in the number of rats that became arrhythmic under LL. We conclude that the light received during lactation affects the strength of the circadian pacemaker and its sensitivity to light. PMID- 12638689 TI - Sex- and age-related temporal variations in intestinal-epithelium proliferation in the suckling mouse. AB - Intestinal-crypt enterocytes are a cell population undergoing constant renewal in the mouse. Both adult and 28 d old animals have been shown to exhibit circadian rhythms in cell proliferative indices, but there are only scant data on the 24 h mitotic activity in the small and large intestine of younger mice. The present studies were thus undertaken in order to characterize the proliferative pattern of enterocytes in the duodenum and colon of 7 and 14 d old males and females of the C3H/S strain. Animals of each sex and from each age group were sacrificed every 4h during a 24 h span, with each animal receiving an injection of colchicine 4h before sacrifice. Samples of duodenum and colon were removed and processed for hematoxylin-eosin staining. Twenty longitudinally sectioned crypts within each sample were analyzed, and the mitotic indices of both cell populations from each animal were estimated. The arithmetic mean +/- SEM for each experimental group were then calculated and the statistical significance of differences between the means assessed by ANOVA and Student t-tests. We observed a greater daily mitotic activity in the duodenum than the colon, and moreover enterocytic proliferation in both those regions was greater in 14 than 7 d old animals. Twenty-four hour variations in mitotic activity occurred in all the experimental groups and tissues except for the large intestine of 7 d old females. Finally, the temporal profile of epithelium proliferation in the suckling mouse varied with age, sex, and site of the intestine studied. PMID- 12638690 TI - The circadian system of Trypanosoma cruzi-infected mice. AB - The effects of Chagas disease on the mammalian circadian system were studied in Trypanosoma cruzi-infected C57-B16J mice. Animals were inoculated with CAI or RA strains of T. cruzi or vehicle, parasitism confirmed by blood specimen visualization and locomotor activity rhythms analyzed by wheel-running recording. RA-strain infected mice exhibited significantly decreased amplitude of circadian rhythms, both under light-dark and constant dark conditions, probably due to motor deficiencies. CAI-treated animals showed normal locomotor activity rhythms. However, in these mice, reentrainment to a 6h phase shift of the LD cycle took significantly longer than controls, and application of 15min light pulses in DD produced smaller phase delays of the rhythms. All groups exhibited light-induced Fos expression in the suprachiasmatic nuclei. We conclude that the main effect of T. cruzi infection on the circadian system is an impairment of the motor output from the clock toward controlled rhythms, together with an effect on circadian visual sensitivity. PMID- 12638691 TI - Serum cortisol levels in goats exhibit seasonal but not daily rhythmicity. AB - The aim of the study was to find out whether there is a daily rhythm in goat serum cortisol concentrations, whether the concentration profiles differ between normal light:dark and constant dark conditions, and whether any seasonal variations might be detected in daily cortisol secretion patterns. Seven Finnish landrace goats were kept at indoor temperature (18-23 degrees C) under artficial lighting that approximately simulated the annual changes of daylength at 60 degrees N. Blood samples were collected for cortisol measurements by radioimmunoassay at 2h intervals during six times of the year: winter (light:dark 6:18h), early spring (10:14h), late spring (14:10h), summer (18:6h), early fall (14:10h), and late fall (10:14h). Cortisol profiles were determined for two consecutive days, first in light:dark (LD) conditions and then in continuous darkness (DD). There was no significant daily rhythm in serum cortisol levels in any time of the year, nor did the profiles in LD and DD conditions show any differences. A significant seasonal variation was, however, detected among the overall cortisol levels. In winter, the concentrations were higher than in any other season, and from early spring to summer they were at their lowest. Under equal photoperiods, the cortisol levels were higher in fall than spring. The difference between winter and summer was confirmed the following year in LD conditions. There was on correlation between serum cortisol and progesterone levels. The results suggest that the possible circadian variation of cortisol secretion in goats is completely masked by external factors, and lighting conditions do not have immediate effects on the daily secretion patterns. The seasonal variation in the overall cortisol levels is most probably related to the changes in photoperiod, because other conditions were relatively constant during the experiment. PMID- 12638692 TI - Chronopathological aspects of disease incidence in rice (Oryza sativa L.). AB - Rice seedlings maintained under uncontrolled glasshouse conditions were inoculated with conidial suspensions of a fungal pathogen, Helminthosporium oryzae, at various times during the 24h. Significant increase in the percent germination and germ tube length of conidia were observed in the rice samples inoculated at 02:00 and 06:00h. The 24 h temporal variation in leaf temperature was positively correlated with variation in stomatal movements. The results indicate a 24 h rhythm in the behavior of the fungal pathogen on the host in relation to the conditions of the growing environment. In all the inoculated seedlings, the appearance of a large number of brown leaf spots was confined to the light span. Among the plants inoculated, earlier initiation of brown leaf spot appearance, maximum number of leaf spots, and highest disease severity were observed when plants were inoculated at 02:00h. There was a positive correlation between disease severity of the host and in vivo values of percent germination of conidia and germ tube length of the pathogen in plants inoculated between 02:00 and 06:00h. The findings of this study implicate that light intensity and temperature could play a predominant role in controlling disease susceptibility rhythms in plants. PMID- 12638693 TI - The relationship between lifestyle regularity and subjective sleep quality. AB - In previous work we have developed a diary instrument-the Social Rhythm Metric (SRM), which allows the assessment of lifestyle regularity-and a questionnaire instrument--the Pittsburgh Sleep Quality Index (PSQI), which allows the assessment of subjective sleep quality. The aim of the present study was to explore the relationship between lifestyle regularity and subjective sleep quality. Lifestyle regularity was assessed by both standard (SRM-17) and shortened (SRM-5) metrics; subjective sleep quality was assessed by the PSQI. We hypothesized that high lifestyle regularity would be conducive to better sleep. Both instruments were given to a sample of 100 healthy subjects who were studied as part of a variety of different experiments spanning a 9-yr time frame. Ages ranged from 19 to 49 yr (mean age: 31.2 yr, s.d.: 7.8 yr); there were 48 women and 52 men. SRM scores were derived from a two-week diary. The hypothesis was confirmed. There was a significant (rho = -0.4, p < 0.001) correlation between SRM (both metrics) and PSQI, indicating that subjects with higher levels of lifestyle regularity reported fewer sleep problems. This relationship was also supported by a categorical analysis, where the proportion of "poor sleepers" was doubled in the "irregular types" group as compared with the "non-irregular types" group. Thus, there appears to be an association between lifestyle regularity and good sleep, though the direction of causality remains to be tested. PMID- 12638694 TI - 24-hour assessment of performance on a palmtop computer: validating a self constructed test battery. AB - A test battery was constructed on a palmtop computer for ambulatory purposes. This study explored whether the test battery could assess circadian rhythmicity underconstant routine conditions. Performance, body temperature, and subjective sleepiness of 12 healthy subjects were measured. The test battery consisted of a sleepiness questionnaire and three performance tests: a vigilance detection test, a working memory test, and a choice-reaction time test. The subjects were divided into early-start and late-start groups and were subjected to the constant-routine protocol. All tests showed a trough in performance in the early morning around 07:00h and a peak in the evening between 21:00 and 23:00h. In addition, an afternoon decrement in performance was observed between 15:00 and 17:00h. On average, the circadian (peak-to-trough) variation of the performance variables amounted to 16.9% +/- 1.7 SEM of the maximum across subjects. The late starters showed a larger impairment in performance during the morning than the early starters. This could be attributed to prior wakefulness. The characteristics of the performance rhythms found in this study replicate findings in several other studies carried out under constant routine conditions. In conclusion, the present test battery appeared to be a good tool for future assessment of performance under natural conditions. PMID- 12638695 TI - Effects of dim or bright-light exposure during the daytime on human gastrointestinal activity. AB - On the basis of our previous findings that bright-light exposure during the daytime has profound influence on physiological parameters such as melatonin secretion and tympanic temperature in humans, we proposed the hypothesis that bright vs. dim light-exposure during the daytime has a different influence on the activity of the digestive system via the endocrine and/or autonomic nervous system. To examine this hypothesis, we conducted a series of counterbalanced experiments in which subjects stayed the daytime (7:00 to 15:00h) under either a dim (80 lux) or bright (5,000 lux) light condition. We measured gastrointestinal activity using a breath hydrogen (indicative of carbohydrate malabsorption) and an electrogastrography (EGG, indicative of gastric myoelectric activity) test. The results showed the postprandial breath hydrogen excretion during the following nighttime period after daytime exposure to the dim-light condition was significantly higher than under the bright-light condition (p < 0.05). In addition, the spectrum total power of the EGG recorded after taking the evening meal was significantly lower for the dim than bright-light condition (p < 0.05). These results support our hypothesis and indicate that dim-light exposure during the daytime suppresses the digestion of the evening meal, resulting in malabsorption of dietary carbohydrates in it. PMID- 12638696 TI - Circadian rhythm of blood pressure challenges office values as the "gold standard" in the diagnosis of gestational hypertension. AB - Despite poor sensitivity and specificity, office blood pressure (BP) determinations are still the "gold standard" for diagnosing gestational hypertension. This prospective blind study evaluates the prognostic value of office values as compared with ambulatory monitoring in pregnancy. We analyzed 2175 BP series systematically sampled from 355 non-preeclamptic pregnant women for 48 h every 4 wks from the first hospital visit until delivery. Women were divided for comparative purposes into three groups: "detected" gestational hypertension, defined on the basis of casual clinical BP> 140/90 mmHg after 20 wks of gestation and hyperbaric index (area of BP excess above the upper limit of a time-specified tolerance interval adjusted for the circadian pattern of the reference population) consistently above the threshold for diagnosing hypertension in pregnancy; "undetected" gestational hypertension, women with office BP < 140/90 mmHg but hyperbaric index consistently above the threshold for diagnosis; and normotension, women with both office values and hyperbaric index below the respective thresholds for diagnosis. Small and insignificant differences in the 24h mean BP between "detected" and "undetected" gestational hypertension is observed in all trimesters, in contrast with highly significant differences between these two groups and normotensive pregnancies. Normotensive women are characterized by highly significant lesser incidence by 60% in preterm delivery, 70% in intrauterine growth retardation, and 50% in delivery by cesarean section (p < 0.001) compared with women with "detected" and "undetected" gestational hypertension (p > 0.715). In pregnancy, the hyperbaric index is markedly superior to office BP measurements for diagnosis of what should be truly considered gestational hypertension, and for prediction of the outcome of pregnancy. PMID- 12638697 TI - Circadian rhythm of accidents in children: a basic activity periodicity. PMID- 12638698 TI - Removal of mercury from clayey soils using electrokinetics. AB - Numerous sites have been polluted with mercury as a result of accidental spills and improper disposal practices, and these mercury-contaminated sites may have adverse effects on human health and the environment. Innovative and cost effective remediation techniques are urgently needed, and this study was performed to investigate the use of electrokinetics for mercury-contaminated soils. Initially, batch tests were performed on two soils, kaolin and glacial till, spiked with mercury(II) to investigate mercury desorption and complexation under different pH environments (pH range 2-12). The complexing agents included disodium ethylenediaminetetraacetate (Na-EDTA), potassium iodide (KI), and sodium chloride (NaCl), and these solutions were used at a concentration of 0.1 M. In addition, deionized water was used for comparison purposes. Based on the batch tests, Na-EDTA and KI were identified as the complexing agents with the greatest potential. The removal efficiency of these complexing agents was then examined by conducting electrokinetic experiments that employed the same solution concentration (0.1 M) and voltage gradient (1.0 VDC/cm) conditions. These tests indicated that for both soils, KI was a more effective complexing agent than Na EDTA under electrokinetics. For the kaolin soil, the electrokinetic treatment using KI removed approximately 97% of the initial contaminant present (500mg/kg of Hg(ll)), leaving a residual concentration of 16 mg/kg of Hg in the soil, whereas on the glacial till soil, KI removed only 56% of the initial contaminant present (500mg/kg of Hg(II)), leaving a residual concentration of 220 mg/kg of Hg in the soil. The lower Hg removal from glacial till is attributed to the presence of organic matter, which increased mercury adsorption or the formation of insoluble mercury surface complexes. PMID- 12638699 TI - Denitrifying characteristics of the multiple stages enhanced biological nutrient removal process with external carbon sources. AB - This research investigated denitrifying activity of activated sludge with three external carbon sources (sodium acetate, methanol and glucose) via a series of batch experiments. Activated sludge used was cultivated in a multiple stages enhanced biological nutrient removal (EBNR) process that exhibited high removal efficiency of effective carbon, nitrogen, and phosphorus. Results showed type of external carbon source had a significant influence on specific nitrate utilization rate, nitrite accumulation, adaptive time of microorganisms, and nitrate removal efficiency. Sodium acetate addition resulted in high phosphate concentration in effluent; meanwhile methanol caused increasing turbidity and carbon breakthrough problem. When glucose was fed to be the external carbon source, accumulative nitrite concentration was higher than that with sodium acetate or methanol addition. When sodium acetate, methanol and glucose were used to be the electron donor, average dosages for nitrate elimination were 6.97, 5.85, and 5.65 mg-COD/mg-N, respectively. Because the final polyhydroxyalkanoates (PHAs) concentrations contained within the biomass were more than the original level and no phosphate re-release was observed, glycogen-accumulating organisms (GAOs) might exist in the multiple stages EBNR process and increased carbon dosage for further nitrate removal. PMID- 12638700 TI - Kinetic responses of activated sludge microorganisms to individual and joint copper and zinc. AB - The individual and joint effects of copper and zinc on the growth kinetics of activated sludge microorganisms were studied in batch reactors fed with glucose as sole carbon source. The observed specific growth rate, substrate utilization rate and removal efficiency were determined at different concentrations of individual and joint Cu and Zn. Activated sludge microorganisms showed different kinetic responses to Cu and Zn, and the Haldane model seemed applicable for the inhibitory effect of Zn, while a first-order kinetics for Cu. Compared to Zn, Cu exhibited high toxicity to activated sludge microorganisms even at a very low concentration of 1.0 mg/dm3. However, Zn concentrations less than 40 mg/dm3 would stimulate the growth of activated sludge microorganism, and Zn acted as inhibitory chemical only at concentrations above this value. The combined effect of Cu and Zn could be regarded as neither synergistic nor antagonistic action. PMID- 12638701 TI - Synergic and competitive inhibition effect between two kinds of arylamines in the formation of hemoglobin adduct in vitro. AB - This article presents our work with HPLC-UV to determine hemoglobin adducts produced by adduction reaction among 4-aminobiphenyl (4-ABP) or/and 2 naphthylamine (2-NA) and human hemoglobin in vitro. The results showed that HPLC UV could efficiently be used to detect aromatic amine-hemoglobin adduct. Two kinds of optimized linear models modified from the traditional dose-response model were screened out. The models implied that synergic effect exists between the two kinds of arylamines adducted with hemoglobin and competitive inhibition effect occurs in the formation of hemoglobin adduct when the two kinds of arylamines are present in the blood synchronously. The results indicate that models for dose-response relationship should take the effects of the other toxicants into consideration in the mixed pollutant system. PMID- 12638702 TI - Fenton-like oxidation of landfill leachate. AB - The treatment of stabilized leachates by means of Fenton's like reagent [Fe(III) H2O2] has been studied. It has been demonstrated that the oxidation state of the catalyst does not influence the efficacy of the process in terms of chemical oxygen demand depletion profiles. The abrupt increase in temperature experienced in oxidation experiments involves a wastage of hydrogen peroxide diminishing the fraction of this reagent addressed at removing COD. If temperature is kept constant, the hydrogen peroxide uptake is 10 mg of H2O2 consumed per mg of COD abated (from 15 to 30 degrees C). Working temperatures above 30 degrees C does not lead to additional COD conversion, contrarily, the percentage of wasted H2O2 is increased. A rough economic analysis of the process indicates that this treatment can be a suitable alternative to deal with this type of effluents. PMID- 12638703 TI - Performance of waste activated carbon as a low-cost adsorbent for the removal of anionic surfactant from aquatic environment. AB - In the present study, different low cost adsorbents were screened for their sodium dodecyl sulfate (SDS, an anionic surfactant) removal capacity. Waste activated carbon (WAC) from the aqua purifier has shown high efficiency for SDS removal. The performance evaluation in the presence of various ions (Ca2+, SO4(2 ), NO3-, and Cl-) and at various pH was studied. Desorption studies were conducted using simple sonication and pH variation technique. Column adsorption studies were performed. SEM and EDS studies were done on the adsorbing material before adsorption, after adsorption and after desorption of SDS. PMID- 12638704 TI - Decolorization of Congo Red with three-dimensional flow-by packed-bed electrodes. AB - The electrochemical removal of the color of the dyestuff Congo Red was investigated experimentally by using packed graphite anodes as well as packed graphite or C-304 stainless steel cathodes in an undivided reactor. The effects of the applied apparent current density, the amount of chloride ion addition, the pH value of the solution, the cathode material, the operating temperature, the volumetric flow rate and the initial concentration of Congo Red on the removal efficiencies were investigated in this study. Experimental results showed that the main pathway of color removal was by the hypochlorite generated in-situ at the anodes. Compared with the indirect oxidation, the color removal due to direct oxidation was insignificant. Cathodic oxidation by using graphite cathodes had little influence on the removal rate in an undivided electrochemical cell. PMID- 12638705 TI - The effect of metal ions on humic acid removal and permeation properties in an ultrafiltration system. AB - With a view to improving the removal of humic acid from aqueous solution, the effect of metal ion addition on the separation of humic acid from water in an utrafiltration (UF) system was investigated. The valence of the metal ion and the molar ratio of humic acid to metal ion strongly affected the permeation flux during ultrafiltration. It was found that the ionic strength, dissociation constant and operating pressure were not major factors affecting the separation performance of the ultrafiltration process. As well as indicating that a suitable ratio of humic acid to metal ion and valence of metal ion were the key factors in improving separation efficiency. The results also showed that separation of humic acid depended on the level of formation of humic acid-metal ion complexes, and on the degree of fouling on the membrane surface. PMID- 12638707 TI - Beryllium in the environment: a review. AB - Beryllium is an important industrial metal because of its unusual material properties: it is lighter than aluminum and six times stronger than steel. Often alloyed with other metals such as copper, beryllium is a key component of materials used in the aerospace and electronics industries. Beryllium has a small neutron cross-section, which makes it useful in the production of nuclear weapons and in sealed neutron sources. Unfortunately, beryllium is one of the most toxic elements in the periodic table. It is responsible for the often-fatal lung disease, Chronic Beryllium Disease (CBD) or berylliosis, and is listed as a Class A EPA carcinogen. Coal-fired power plants, industrial manufacturing and nuclear weapons production and disposal operations have released beryllium to the environment. This contamination has the potential to expose workers and the public to beryllium. Despite the increasing use of beryllium in industry, there is surprisingly little published information about beryllium fate and transport in the environment. This information is crucial for the development of strategies that limit worker and public exposure. This review summarizes the current understanding of beryllium health hazards, current regulatory mandates, environmental chemistry, geochemistry and environmental contamination. PMID- 12638706 TI - Mapping air contaminant concentrations using remote sensing FTIR. AB - Spatial Mapping of air contaminant concentrations on a two-dimensional plane in outdoor air using remote sensing FTIR was conducted in this study. The quasitime stationary experiment was carried out after chloroform and methanol dispersed 200 min later freely. The Path-Integrated Concentrations (PIC) in different optical path directions were obtained simultaneously. The kriged maps as well as the two dimensional contour maps from PIC data of chloroform and methanol were plotted and compared. There was a consistency between the concentration peak positions in the two graphs. Although the concentrations were low as a whole, the kriged map of chloroform had a sharper peak than that of the methanol. It can be concluded from the experiment that mapping contaminant concentrations could help understand the peak locations of industrial emissions and provide improved estimates of pollutant flow and distribution. This study had the guiding significance in environmental monitoring and protection. PMID- 12638708 TI - Opportunities for designing health insurance demonstrations. PMID- 12638709 TI - Nonprofit health systems: a promising new class of corporate citizen. AB - Leading nonprofit health systems are demonstrating that communities can benefit from the emergence of this new class of corporate citizens. Just as the business sector has produced many good corporate citizens to the great advantage of American society, health systems with sufficient financial and organizational gravitas increasingly are assuming these roles, and in so doing, are making positive differences in their communities. More system leaders, however, must find compelling reasons to assume these demanding roles. They also must learn how to play citizenship roles more fully and effectively if the potential social good available through health systems' corporate citizenship is to be realized. PMID- 12638710 TI - An examination of factors in the withdrawal of managed care plans from the Medicare+Choice program. AB - Participation of health maintenance organizations (HMOs) in the Medicare+Choice program, expected to rise rapidly after passage of the Balanced Budget Act of 1997, has gone in just the opposite direction. Because plans have left in such large numbers, Congress has taken remedial measures to remove restrictions and increase payments. To date these efforts have failed. This paper uses plan organizational characteristics, market position, and financial performance to quantify the reasons why some HMOs exited at the end of 1998. The findings suggest HMO participation in Medicare+Choice will continue to fall unless major changes are made to the overall Medicare program and the method of paying HMOs. PMID- 12638711 TI - Testing the effect of quality reports on the health plan choices of Medicare beneficiaries. AB - This article describes a laboratory experiment that used a convenience sample of 225 Medicare beneficiaries to test the effects of comparative quality information on plan choice. Providing information about quality did not significantly influence the choice between Original Medicare and a health maintenance organization (HMO), but did affect the choice of HMO. Results from this experiment suggest that information about plan quality may not be effective in encouraging beneficiaries to leave Original Medicare and join HMOs that are rated high in quality. Furthermore, beneficiaries choosing among HMOs were not inclined to select a low-cost HMO, even when it was rated higher in quality. Implications for policy are discussed. PMID- 12638712 TI - Welfare leavers' use of Medicaid transitional medical assistance in California, 1993-1997. AB - Using administrative and survey data, we estimate participation rates in Transitional Medical Assistance (TMA) for the period 1993-97 by California welfare leavers during their first six months post-welfare. We find that although many welfare leavers were eligible for TMA (35% to 47% of exiters), only 26% of eligible people were enrolled in the TMA program. Another 14% were covered by non TMA Medicaid for the entire six months. Most TMA-eligible exiters had Medicaid coverage (all of it non-TMA) for less than six months (49%) or no Medicaid coverage at all (11%). Supplementary analyses using data from the National Survey of America's Families indicate that if fully implemented, TMA could have substantially reduced uninsurance among welfare leavers. PMID- 12638713 TI - Ownership and changes in hospital inefficiency, 1986-1991. AB - This study examines how ownership affected changes in hospital inefficiency after the introduction of prospective payment by Medicare. Using a national data set, we estimate cost frontiers for 1986 and 1991 to assess hospitals' efficiency relative to best practice in both those years. We then use regression analysis to determine the effect of ownership on the change in hospitals' efficiency. The results indicate that, in both 1986 and 1991, mean inefficiency was highest for for-profit hospitals and lowest for not-for-profit hospitals, with government hospitals falling in the middle. Moreover, between 1986 and 1991, both for-profit and government hospitals had significantly less improvement in efficiency than not-for-profit hospitals, all else equal. PMID- 12638714 TI - How many hospital beds? AB - For many years, average bed occupancy level has been the primary measure that has guided hospital bed capacity decisions at both policy and managerial levels. Even now, the common wisdom that there is an excess of beds nationally has been based on a federal target of 85% occupancy that was developed about 25 years ago. This paper examines data from New York state and uses queueing analysis to estimate bed unavailability in intensive care units (ICUs) and obstetrics units. Using various patient delay standards, units that appear to have insufficient capacity are identified. The results indicate that as many as 40% of all obstetrics units and 90% of ICUs have insufficient capacity to provide an appropriate bed when needed. This contrasts sharply with what would be deduced using standard average occupancy targets. Furthermore, given the model's assumptions, these estimates are likely to be conservative. These findings illustrate that if service quality is deemed important, hospitals need to plan capacity based on standards that reflect the ability to place patients in appropriate beds in a timely fashion rather than on target occupancy levels. Doing so will require the collection and analysis of operational data-such as demands for and use of beds, and patient delays--which generally are not available. PMID- 12638715 TI - Adverse selection and generosity of alcohol treatment benefits. AB - Concerns about attracting disproportionate numbers of employees with alcohol problems limit employers' willingness to offer health plans with generous alcohol treatment benefits. This paper analyzes two potential avenues of adverse selection, namely biased enrollment into plans and biased exit from plans offered by 57 employers between 1991 and 1997. We compare alcohol treatment use rates and costs of new and old enrollees between more generous and less generous plans; we also analyze disenrollment rates and enrollment duration by plan generosity for users and nonusers of alcohol treatment services. To avoid confounding benefit generosity with other plan features, in particular the use of managed care mechanisms, we compare plans that were administered in the same way by a large managed behavioral health care organization. Overall, we find no evidence of adverse selection into more generous plans. Contrary to the selection hypothesis, treatment costs of new members compared to old members are lower in firms with more generous treatment benefits than in firms with more limited benefits. Also, users of alcohol treatment services do not remain disproportionately enrolled longer in plans with generous benefits. PMID- 12638716 TI - Amplification-based nucleic acid scanning techniques to assess genetic polymorphism in Candida. AB - Opportunistic pathogen Candida causes common fungal infections that manifest both superficially and systemically, especially in compromised patients. Although C. albicans is by far the main etiological agent of candidosis, the frequency of isolation of other non-albicans species such as C. glabrata and C. krusei is increasing at an alarming rate. Therefore, the epidemiology, pathogenicity, and diagnosis of infections due to these organisms are of great importance. Of a variety of genotyping methods utilized for strain delineation of these Candida species, amplification-based techniques such as randomly amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), restriction digestion mediated PCR (RFLP-PCR), and single-stranded conformational polymorphism (SSCP) and microsatellite PCR (interrepeat PCR, IR-PCR) are the most popular and widely used. In the last decade or so these techniques have helped unravel the clinical epidemiological features of pathogenicity, diversity, microevolution, and natural heterozygosity in Candida species. Here we review in detail the basic principles of RAPD, the nature of the primer and factors influencing its selection, and the limitations of RAPD assays as well as analysis and interpretation of banding profiles generated using the software programs. In addition, the principles of other RAPD-based amplification techniques (AFLP, RFLP-PCR, SSCP, and IR-PCR) and their application in molecular epidemiologic studies of Candida species in particular and other fungi in general are also reviewed. It is concluded that these methods have wide applicability in genotyping fungi, although they differ greatly in their resolution and have advantages and drawbacks depending on the task in question. PMID- 12638717 TI - The directed mutation controversy in an evolutionary context. AB - Neo-Darwinists have long held that random mutations produce genetic differences among individuals, and selection increases the frequency of advantageous alleles. In 1988, Cairns et al. claimed that an environmental pressure can cause advantageous mutations to occur in specific genes to alleviate that particular pressure. Directed mutation, as proposed by Cairns, has been all but eradicated from evolutionary thinking. However, more than a decade of research spurred by the Cairns et al. paper has cast doubt on three neo-Darwinian principles: (1) mutations occur independently of the environment, (2) mutations are due to replication errors, and (3) mutation rates are constant. This mini-review explores the history of the controversy and the decade of research that followed so as to place it in an evolutionary context. Several of the cellular mechanisms and models that explain the increased genetic diversity in populations experiencing adverse environmental pressure are described. In most cases it is clear that the increased genetic diversity is due to breakdowns of cellular machinery or alleles evolved for a purpose other than increasing genetic diversity, rather than to cellular systems that have been evolutionarily selected to increase the genetic diversity in times of stress. PMID- 12638718 TI - Increasing diversity within Chlamydiae. AB - In recent years, 16S ribosomal DNA analyses has allowed the recognition of new chlamydia organisms, requiring the creation of new species, genera, and families within this unique, deep lineage of prokaryotes. The trachoma and psittaci groups chlamydiae are now recognized as separate genera, Chlamydia and Chlamydophila, respectively, and biovars of each group have been elevated to the species rank. Simkania and Parachlamydia have been associated with human respiratory infections, while Waddlia seems to be implicated in abortion in bovins. DNA amplification studies targeting the 16S rDNA have revealed a richer diversity within chlamydiae, identifying new lineages from both environmental and clinical samples. Further studies will be of interest to both examine the ecology and evaluate the clinical importance of these novel chlamydiae. Herein, we provide a summary of literature and our data about the novel chlamydial lineages. PMID- 12638720 TI - Isolated familial somatotropinomas: clinical features and analysis of the MEN1 gene. AB - Isolated familial somatotropinomas (IFS) rarely occurs in the absence of multiple endocrine neoplasia type I (MEN1) or the Carney complex. In the present study we report two Italian siblings affected by GH-secreting adenomas. There was no history of parental consanguinity. The sister presented at 18 years of age with secondary amenorrhea and acromegalic features and one of her two brothers presented with gigantism at the same age. Endocrinological investigations confirmed GH hypersecretion in both cases. Although a pituitary microadenoma was detected in both patients, transsphenoidal surgery was not successful. The sister received conventional radiotherapy and acromegaly is now considered controlled; the brother is being treated with octreotide LAR 30 mg monthly and the disease is considered clinically active. Patients, their parents and the unaffected brother underwent extensive evaluation, and no features of MEN1 or Carney complex were found. Analysis of polymorphic microsatellite markers from chromosome 11q13 (D11S599, D11S4945, D11S4939, D11S4938 and D11S987) showed that the acromegalic siblings had inherited different maternal chromosomes and shared the paternal chromosome. No pathogenic MEN1 sequence changes were detected by sequencing or dideoxy fingerprinting of the coding sequence (exons 2-10) and exon/intron junctions. Although mutations in the promoter, introns or untranslated regions of the MEN1 gene cannot be excluded, germline mutations within the coding region of this gene do not appear responsible for IFS in this family. PMID- 12638719 TI - Expression of aromatase P450 is increased in spontaneous prolactinomas of aged rats. AB - We have recently reported the presence of aromatase P450 in the rat hypophysis. This enzyme is responsible for the aromatization of testosterone to estradiol. Since the induction of prolactinomas has been demonstrated in the rat following chronic treatment with estradiol, the aim of the present study was to analyze whether a relationship exists between the presence of pituitary aromatase and the appearance of spontaneous prolactinomas in aged rats. Of a series of 90 adenomas studied, 53% showed prolactin immunoreactive cells and were classified as prolactinomas; only 33% of the adenomas were pure prolactinomas and the other 20% were multi-hormonal protactinomas. Moreover, 60% of the adenomas were aromatase positive tumors. Interestingly, 100% of the pure prolactinomas were aromatase positive while only 60% of the multi-hormonal prolactinomas expressed the enzyme. Western blotting with anti-aromatase antibodies revealed a 3.8-fold increase in expression of aromatase in pituitary tumors as compared to normal rat pituitary gland. Double immunohistochemical labeling detected the coexistence of prolactin and aromatase P450 in prolactinoma cells. ACTH- and LH-positive adenomas were considered as controls; only multi-hormonal ACTH and LH tumors display aromatase positive cells and all of these also contained prolactin-positive cells. Our results demonstrate for the first time that aromatase is expressed in pituitary adenomas and that it is related to the functional nature of the tumor, especially in the case of pure prolactinomas, suggesting the possibility that an abnormally high conversion of testosterone into estradiol in pituitary cells may contribute to the genesis of spontaneous prolactinomas in aged rats. PMID- 12638721 TI - Self image and quality of life in acromegaly. AB - Chronic effects of excessive GH secretion are not only disabling but also disfiguring. Most acromegalic patients present with 4-10 years history of changes in features, bony overgrowth, soft tissue swelling, skin changes, diabetes mellitus, hypertension and other cardiovascular symptoms. In addition to sign and symptoms of GH excess the patients demonstrate a loss initiative and spontaneity, mood swings, an impairments in self esteem, body image distortion, disruption in interpersonal relations and social withdrawal anxiety. Little research is available on the psychological features of acromegaly, despite their importance in this chronic and debilitating condition. Extremely little, however has appeared in the medical and psychological literature about mental and emotional disfunction in patients with acromegaly. PMID- 12638722 TI - Oncological complications of excess GH in acromegaly. AB - Around fifteen percent of the deaths reported in acromegaly are attributable to malignancy (SM Melmed, J Clin Endocrinol Metab 2001;86:2929-2934; A Mestron, SM Webb, In: Endocrine Society, San Francisco, June 2002 (abstract)), uncontrolled disease is associated with a growth advantage for concurrent neoplasms, which are more likely to be aggressive; however, there is no clear evidence of de novo cancer initiation in acromegaly and no proven causal relationship of acromegaly with malignant disease. Overall and cancer mortality in acromegaly have been shown to correlate with the degree of GH control; if post therapy GH is controlled, both the overall and cancer mortality do not appear to differ from that of the normal population (SM Orme et al., J Clin Endocrinol Metab 1998;83:2730-2734; JD Nabarro, Clin Endocrinol 1987;26:481-512). However, no long term prospective evaluation of cancer prevalence or its relation to biochemical or clinical disease activity is available. IGF-1 appears to exert a permissive effect on tumorigenesis; there is no clear evidence that tumor initiation is triggered by IGF-1 in acromegaly; nevertheless, since IGF-1 may be higher in neoplasms, aggressive treatment aimed at controlling the disease activity will theoretically be advantageous for acromegalic patients with cancer. The prevalence of colon polyps and colon cancer appear to be incresed in acromegaly as is mortality for colon cancer (Orme et al., 1998), so regular colonoscopy screening and polypectomy would seem advisable, especially in older patients with active acromegaly. Surveillance for prostate cancer in elderly males with high IGF-1, especially if also receiving testosterone replacement therapy, is recommendable, by measurement of serum PSA, rectal examination and/or prostatic ultrasound. In women, mammography should be offered, especially after the age of 50 years, as in the normal population. Neither prostate nor breast cancer have been consistently shown to have an increased prevalence in acromegaly, but larger prospective epidemiological studies are required to study this further. PMID- 12638724 TI - Hypothalamic-pituitary sarcoidosis with hypopituitarism. Long-term remission with methylprednisolone pulse therapy. AB - A 29-year-old man presented with diplopia and decreased strength in the left arm. A magnetic resonance image (MRI) showed an extensive hypothalamic and pituitary gland mass, and hormonal studies showed partial hypopituitarism and mild hyperprolactinemia without diabetes insipidus. Biopsies of the hypothalamic lesion and of a mediastinal lymph node demonstrated noncaseating granulomas, and a Kveim-Siltzbach test was positive. He was successfully treated with a regimen of high-dose intravenous methylprednisolone pulse therapy for eight weeks along with a low dose of oral corticosteroids which was maintained indefinitely. An MRI obtained immediately after pulse therapy revealed a substantial reduction in the hypothalamic-pituitary mass, which was maintained in an MRI performed 3 years later. However, hormonal deficits persisted and indefinite hormonal substitutive therapy was required. PMID- 12638723 TI - Therapy of diabetes and dyslipidemia in acromegaly. PMID- 12638725 TI - A relapsing remitting lymphocytic hypophysitis. AB - Lymphocytic Hypophysitis is a rare autoimmune disease of the pituitary presenting mainly with features of a mass lesion and loss of pituitary function. Its course is quite unpredictable and its treatment is still controversial as experience in the field remains scarce. We describe a 45 year-old woman with a history of recurrent fever and meningeal symptoms who was referred 3 years later to our department for pituitary insufficiency. Hormonal studies revealed an anterior pituitary deficiency and autoimmune thyroiditis. Pituitary hypertrophy as evidenced by magnetic resonance imaging showed complete regression upon hydrocortisone substitution therapy. Two years later, fever and meningeal symptoms recurred as well as pituitary hypertrophy. Cerebrospinal fluid analysis revealed an aseptic lymphocytic meningitis. Pituitary biopsy confirmed the diagnosis of LH. The patient was started on prednisone 70 mg per day. She improved clinically and radiologically and remained free of symptoms thereafter. At a recent follow up the MRI showed an empty sella turcica. Hereby we illustrate a yet unreported pattern of LH presenting as a recurrent aseptic meningitis. We also describe one of the long-term course of the disease which is still unpredictable. PMID- 12638726 TI - Mitochondrial encephalomyopathies. AB - Mitochondrial encephalomyopathies are diseases caused by defective oxidative phosphorylation (OXPHOS), and affect the nervous system and/or skeletal muscle. They have emerged as a major entity among the neurometabolic diseases of childhood with an incidence of 1 in 11,000 children, and also have a high prevalence in adults. The first pathogenic mutation of human mitochondrial DNA (mtDNA) was discovered in 1988. Since then more than 100 mutations of mtDNA have been reported, including point mutations of genes encoding transfer RNA, ribosomal RNA, and proteins, as well as large-scale deletions. The first nuclear DNA gene mutation causing OXPHOS disease was described in 1995. Mutations in nuclear genes may affect the respiratory chain by various mechanisms. Pathogenic mutations of nuclear-DNA-encoded subunits of complex I and II have been demonstrated as have mutations of respiratory chain assembly proteins. Several nuclear genes associated with mtDNA maintenance have been found to be associated with mitochondrial disorders since mutations in these genes predispose to multiple mtDNA deletions and/or reduced copy number of mtDNA. The genotype phenotype correlation is not yet entirely clear, but new animal models will enhance our ability to study the pathophysiology of OXPHOS disorders. PMID- 12638727 TI - The pleiotropic effects of erythropoietin in the central nervous system. AB - Erythropoietin (Epo) is a hydrophobic sialoglycoproteic hormone produced by the kidney and responsible for the proliferation, maturation, and differentiation of the precursors of the erythroid cell line. Human recombinant erythropoietin (rHuEpo) is used to treat different types of anemia, not only in uremic patients but also in newborns with anemia of prematurity, in patients with cancer-related anemia or myeloproliferative disease, thalassemias, bone marrow transplants, or those with chronic infectious diseases. The pleiotropic functions of Epo are well known. It has been shown that this hormone can modulate the inflammatory and immune response, has direct hemodynamic and vasoactive effects, could be considered a proangiogenic factor because of its interaction with vascular endothelial growth factor, and its ability to stimulate mitosis and motility of endothelial cells. The multifunctional role of Epo has further been confirmed by the discovery in the central nervous system of a specific Epo/Epo receptor (EpoR) system. Both Epo and EpoR are expressed by astrocytes and neurons and Epo is present in the cerebrospinal fluid (CSF). Therefore, novel functions of Epo, tissue-specific regulation, and the mechanisms of action have been investigated. In this review we have tried to summarize the current data on the role of Epo on brain function. We discuss the different sites of cerebral expression and mechanisms of regulation of Epo and its receptor and its role in the development and maturation of the brain. Second, we discuss the neurotrophic and neuroprotective function of Epo in different conditions of neuronal damage, such as hypoxia, cerebral ischemia, and subarachnoid hemorrhage, and the consequent possibility that rHuEpo therapy could soon be used in clinical practice to limit neuronal damage induced by these diseases. PMID- 12638728 TI - Beta-amyloid precursor protein staining in nonhomicidal pediatric medicolegal autopsies. AB - Immunohistochemical staining for beta-amyloid precursor protein (betaAPP) has been validated as a marker for axonal injury in adults surviving > or = 2 hours after white matter damage. The significance of betaAPP staining in pediatric brains and spinal cords is not as well established. We evaluated the white matter immunoreactivity for betaAPP from a variety of pediatric medicolegal autopsies: natural disease (non-Sudden Infant Death Syndrome [SIDS]), SIDS, motor vehicle accidents, drowning, near-drowning, overlay, carbon monoxide toxicity, miscellaneous trauma, and mechanical asphyxia. The cases of carbon monoxide toxicity, motor vehicle accidents (death at scene), drowning (with resuscitation), and a natural (non-SIDS) death had no significant white matter staining. The traumatic deaths with a significant survival interval, a variety of natural deaths, the near-drowning case, and surprisingly, all SIDS had detectable betaAPP white matter immunostaining. These results demonstrate that features other than traumatic axonal injury, such as metabolic insults and hypoxic ischemic injury secondary to vascular compromise, must contribute to betaAPP immunostaining. In addition, we describe a variety of betaAPP-immunoreactive structures not previously reported in the pediatric population. This study illustrates that betaAPP immunostaining enhances detection of a variety of white matter changes, and provides a basis for interpretation of these results. PMID- 12638729 TI - Beta-amyloid racemized at the Ser26 residue in the brains of patients with Alzheimer disease: implications in the pathogenesis of Alzheimer disease. AB - Oligomeric and fibrillar beta-amyloid (Abeta) may be toxic in Alzheimer disease (AD), especially after post-translation modification cumulative over time. Racemization of Ser and Asp residues of Abeta in senile plaques (SPs) occurs as an age-dependent process in AD. We previously reported that Abeta1-40 racemized at Ser26 is soluble and susceptible to proteolysis yielding toxic [D Ser26]Abeta25-35/40 fragments in vitro and in vivo. Here, we focus on the localization of racemized Ser26 residues in AD brains within the limbic system, the earliest site of AD histopathology. We developed antisera (20.1 and 22.7). each with epitopes within [D-Ser26]Abeta25-40. Two forms of truncated [D Ser26]Abeta were detected either in SPs or within neurons in all 11 AD-affected brains, but not in age-matched controls. [D-Ser26]Abeta25/26-35 (detected by 20.1) was localized to plaque cores, extracellular neurofibrillary "ghost" tangles and vascular amyloid deposits. In contrast, [D-Ser26]Abeta25-40 (detected by 22.7) was observed in most neurons containing intracellular neurofibrillary tangles, but not in SPs. These results suggest [D-Ser26]Abeta]1-40, formed during aging, becomes soluble and diffuses from SPs. It is then proteolyzed to [D Ser26]Abeta25-35/40, which is toxic and may contribute to the neurodegeneration. This hypothesis may explain the long lag between SP formation and neurofibrillary degeneration in AD brains. PMID- 12638730 TI - Insulin deficiency rather than hyperglycemia accounts for impaired neurotrophic responses and nerve fiber regeneration in type 1 diabetic neuropathy. AB - Diabetic polyneuropathy (DPN) shows more severe functional and structural changes in type 1 than in type 2 human and experimental diabetes. We have previously suggested that these differences may be due to insulin and/or C-peptide deficiencies in type 1 diabetes. To further explore these differences between type I and type 2 DPN, we examined factors underlying nerve fiber regeneration in the hyperinsulinemic type 2 BB/Z-rat and compared these with previous data obtained from the iso-hyperglycemic, insulin and C-peptide-deficient type 1 diabetic BB/Wor-rat. The expression of neurotrophic factors and cytoskeletal proteins were studied in L4 and L5 dorsal root ganglia (DRG) at various time points after sciatic nerve crush. The data were compared to those of nondiabetes prone BB-rats. Insulin-like growth factor 1 (IGF-1) and TrkA levels were lower in DRG from type 1 than from those of type 2 and control BB-rats. On the other hand, IGF-1 receptor expression was increased at baseline in type 1 BB/Wor-rats and decreased after crush injury, whereas its expression increased after crush injury in both control and type 2 BB/Z-rats. Following crush injury, betaII- and betaIII tubulins were upregulated in type 2 BB/Z and control rats, which did not occur in type 1 BB/Wor-rats. Furthermore, type 2 BB/Z-rats showed the normal downregulation of low and medium molecular neurofilament (NF-L and NF-M, respectively), which did not occur in type 1 BB/Wor-rats. These findings were associated with significantly milder abnormalities in axonal elongation and caliber growth of regenerating fibers in type 2 compared to type 1 diabetic rats. These data suggest that impaired insulin signaling in type 1 diabetic nerve may be of greater significance in the regulation of neurotrophic and neurocytoskeletal protein synthesis than hyperglycemia in explaining the differences in nerve fiber regeneration between type 2 and type 1 diabetes. PMID- 12638731 TI - There is differential loss of pyramidal cells from the human hippocampus with survival after blunt head injury. AB - The experimental literature has shown that neurons within sub-fields of the hippocampus possess differential sensitivities to cell loss after different types of insult to the brain. In humans, after blunt head injury, differential neuronal responses between sub-fields of the hippocampus up to 72 hours after injury have been documented. But, in only a small part of the literature have data for alterations in real numbers of neurons been provided. In this study the hypothesis was tested that, after severe blunt head injury in humans, the total number of neurons within a defined volume of brain tissue differed between different sub-fields of the hippocampus and between groups of patients with differing post-traumatic survivals. Stereological methods were used to measure total cross-sectional area of sub-fields of the hippocampus taken at the level of the lateral geniculate nucleus and count numbers of neurons within each of the CA1, CA2, CA3, and CA4 sub-fields of the hippocampus in patients. The patients used in this study were categorized as follows: Group 1 (early) had survived for 1 week or less; Group 2 (late) survived 6 months or longer after fatal severe head injury; and Group 3 (controls) consisted of age-matched patients that had no history of head injury or disease prior to death. There was a significant loss in cross-sectional area in sub-fields CA3 and CA4 at 1 week or less after injury and in sub-field CA1 at 6 months and greater survival. There was no change in CA2. There was loss of neurons from within a predefined volume of brain tissue in sub fields CA1, CA3, and CA4 one week or less after injury. But there was no loss in CA2. There was continued loss of neurons from sub-fields CA1 and CA4 between 1 week and 6 months and greater survival, but there was no loss of neurons in sub fields CA2 and CA3 within the same period. These novel data show that after human severe head injury there is first an acute loss (1 week or less survival) of pyramidal neurons in all hippocampal sub-fields except CA2. Second, there is an ongoing loss of neurons in sub-field CA1 and, most notably, in sub-field CA4, in patients surviving for more than 6 months. However, in neither group of patients is there loss of neurons from sub-field CA2. PMID- 12638732 TI - Increased vulnerability to focal ischemic brain injury in human apolipoprotein E4 knock-in mice. AB - Accumulating evidence suggests that among the 3 human apolipoprotein E (apoE) isoforms encoded by the human APOE gene, the e4 allele may act to exacerbate brain damage in humans and animals. This study aimed to compare the isoform specific vulnerability conferred by human apoE to ischemic brain damage, using mice expressing human apoE isoforms (apoE2, apoE3, or apoE4) in place of mouse apoE, produced by the gene-targeting technique in embryonic stem cells (knock-in, KI). Homozygous human apoE2 (2/2), apoE3 (3/3), or apoE4 (4/4) KI mice were subjected to permanent focal cerebral ischemia by a modified intraluminal suture method. Twenty-four h thereafter, brain damage, (as estimated by infarct volume and neurologic deficit) was significantly worse in 4/4 KI mice versus 2/2 or 3/3 KI mice (p < 0.001 for each comparison), with no significant differences between 2/2 and 3/3 KI mice. Immunohistochemistry for human apoE expression revealed similar apoE distribution with no significant difference in the immunostaining intensity among the 3 lines of KI mice. Notably. increased expression of human apoE was detected in neurons and astrocytes in the peri-infarct area, and a punctate expression pattern was evident in the border between the infarct and peri-infarct areas in all KI mice subjected to ischemia. Taken together, our results show that apoE affects the outcome of acute brain damage in an isoform specific fashion (apoE4 > apoE3 = apoE2) in genetically engineered mice. PMID- 12638733 TI - Posthemorrhagic ventricular dilation in the neonate: development and characterization of a rat model. AB - Intraventricular hemorrhage is a common complication of prematurity. Posthemorrhagic ventricular dilation (PHVD) has a high rate of disability and no safe and effective treatment. Its pathogenesis is poorly understood, largely because of the lack of a satisfactory animal model. We have developed a model of neonatal PHVD in the rat. Seven-day-old (P7) Wistar rat pups were given 80-microl injections of citrated rat blood or artificial cerebrospinal fluid (CSF) into alternate lateral ventricles on P7 and P8. Intracranial pressure was monitored and increased briefly by over 8-fold. Some rats received further 10-microl intraventricular injections of India ink on P21. Animals were weighed daily and simple neurologic tests performed. On P21 (or P22 if India ink had been injected), the rats were perfusion-fixed and blocks processed for paraffin histology. Sixty-five percent of pups injected with blood and 50% injected with artificial CSF developed dilated lateral ventricles, with patchy loss of ependyma, marked astrocytic gliosis, and rarefaction of periventricular white matter. India ink injection revealed slow transit of CSF from the dilated lateral ventricles but eventual passage into the subarachnoid space. Pups that had received intraventricular injections but did not develop ventricular dilation nonetheless had lighter brains than littermate controls (p < 0.001). Body weights were not significantly different from controls. Hydrocephalic animals had reduced motor performance as assessed by a grip traction test (p = 0.0002). This model is well suited to studying the pathogenesis of PHVD. PMID- 12638734 TI - Changes in neuronal migration in neocortex of connexin43 null mutant mice. AB - To identify a neural phenotype in connexin43 null mutant mice, electrophysiological properties, intercellular communication and neuronal migration were studied in the developing neocortex. In acute slice preparations from newborn mice, electrophysiological characteristics of cortical and hippocampal neurons were not significantly different between wild type and null mutant mice. However, gap junctional coupling as assessed by fluorescence recovery after photobleaching was significantly attenuated in neocortical brain slices of null mutant mice. To assess neuronal migration, dividing cells were labeled with bromodeoxyuridine (BrdU) on embryonic days 12, 14 and 16, respectively, corresponding to the period of cortical neurogenesis, and the neocortex examined 2 or 3 days after the labeling. BrdU-labeled cells were distributed in the neocortical wall with a significant change in the pattern in the neocortex of the null mutant, where labeled cells accumulated in the intermediate zone or in the inner part of the cortical plate. The result suggests a significant delay in neocortical neuronal migration in the connexin43 null mutants, and a possible role of connexin43 in this process through yet unidentified mechanisms. PMID- 12638735 TI - Metallothionein-I overexpression decreases brain pathology in transgenic mice with astrocyte-targeted expression of interleukin-6. AB - Transgenic expression of interleukin-6 (IL-6) in the CNS under the control of the glial fibrillary acidic protein (GFAP) gene promoter (GFAP-IL6 mice) causes significant damage and alters the expression of many genes, including a dramatic upregulation of metallothionein-I (MT-I). The findings in this report support the idea that the upregulation of MT-I observed in GFAP-IL6 mice is an important mechanism for coping with brain damage. Thus, GFAP-IL6 mice that were crossed with TgMTI transgenic mice (GFAP-IL6xTgMTI) and overexpressed MT-I in the brain showed a decreased upregulation of cytokines such as IL-6 and a diminished recruitment and activation of macrophages and T cells throughout the CNS but mainly in the cerebellum. The GFAP-IL6 mice showed clear evidence of increased oxidative stress, which was significantly decreased by MT-I overexpression. Interestingly, MT-I overexpression increased angiogenesis in GFAP-IL6 mice but not in control littermates. Overall, the results strongly suggest that MT-I+II proteins are valuable factors that protect against cytokine-induced CNS injury. PMID- 12638736 TI - Increased neuronal nitric oxide synthase expression in the gracile nucleus of brainstem following electroacupuncture given between cutaneous hindlimb acupuncture points BL 64 & BL 65 in rats. AB - This study was to examine the influence of electroacupuncture (EA) stimulation on neuronal nitric oxide synthase (nNOS) expression in the brainstem nuclei in rats. Low-frequency EA stimulation (3 pulses/sec) was applied between 2 acupuncture points (acupoints), Jinggu (BL 64) and Shugu (BL 65) which are cutaneously located at hindlimb, in rats anesthetized with ketamine. After 2 hours stimulation and sham-treatment, animals were perfused with 4% paraformaldehyde. Sections of rat medulla were examined by immunolabeling with a polyclonal antibody directed against nNOS. The brainstem nuclei were also visualized by NADPH-diaphorase histochemistry, a marker of nNOS activity. nNOS levels were quantified by using a microscope with reticule grid to count the number of positive cells over an area. Unilateral EA stimulation of BL 64 and BL 65 in rats caused increases in nNOS immunostaining cells in the ipsilateral and contralateral gracile nucleus compared with sham-treated rats (P<0.05, n=6). NADPH-diaphorase positive cells were also increased in the gracile nucleus of the rats with EA stimulation. Neither nNOS immunostaining nor NADPH-diaphorase reactivity was altered in the nucleus tractus solitarius, rostral ventral medull and other brainstem nuclei in rats with EA stimulation. These results show that nNOS immunoreactivity and NADPH-diaphorase reactivity are consistently increased in the gracile nucleus by low-frequency EA applied to BL 64 and BL 65. We conclude that EA stimulation of the cutaneous hindlimb acupoints induces nNOS expression in the gracile nucleus, and enhanced nNOS-NO in the area may mediate somatosympathetic reflex activities, which contribute to therapeutic effects of acupuncture. PMID- 12638737 TI - Is acupuncture an useful tool for pain-treatment in ophthalmology? AB - Pain that does not respond to conventional treatment procedures makes it necessary to look for alternative methods. Acupuncture is an ancient procedure with empirical effects on pain. Previous studies established the increased output of messengers at neuronal junctions in spinal cord and hypothalamic locations, especially of endorphins which inhibit the perception of pain. We treated several painful symptoms with acupuncture and evaluated the outcome of the treatment. Patients with various kinds of therapy-refractory pain and patients in whom conventional treatment methods could not be applied were included in the study. The diagnoses included glaucoma. Tolosa-Hunt-Syndrome, ophthalmic migraine, blepharospasm, and dry eyes. In one case acupuncture was used for analgesia during surgery. Acupuncture was performed with sterile disposable needles, at points known to have an empirical analgesic effect. The stimulation was adapted to the patient's individual needs. VAS assessments before and after acupuncture were compared. The t-test was used for statistical evaluation. Acupuncture had no side effects, but reduced pain to a variable extent. Especially in cases of severe pain and in surgery, very effective pain reduction was achieved. In general, pain was significantly reduced in all patients by the use of acupuncture. A statistically significant effect was noted (p < 0.05). Further studies should be conducted to demonstrate the specific effect in larger patient populations. Monitoring neurotransmitter activity will possibly help to illustrate the effect. PMID- 12638738 TI - Differential expression of age-related genes in the cerebrum of senescence accelerated mouse (SAMP10) and analysis of acupuncture interference using DD-PCR technique. AB - In this paper, we applied DD-PCR (differential display polymerase chain reaction) technique to analyze the differential expression of age-related genes in the cerebrum of senescence-accelerated mouse (SAMP10) and effect of acupuncture on them. The results showed that the expression levels of some genes either increased or decreased with increasing aging, it implies that they were probably related to accelerated senescence of SAMP10. Compared with the control group at the matched age, the expression levels of the genes of cerebrum of acupunctured mice were all affected after acupuncturing acupoints, the genes that over express with aging were down-regulated (acupuncture decreases the expression level of related genes) but that less express were up-regulated (acupuncture increases the expression level of related genes). These changes all showed a tendency towards expression level of the young mice. We observed for the first times at the transcriptional levels that the effect of acupuncture is the adjustment that probably interferes with the senescence process from whole body and proved also the acupoints have specificity of treating compared with non-acupoints. PMID- 12638739 TI - Effect of electroacupuncture and transcutaneous electrical nerve stimulation at Hegu (LI.4) acupuncture point on the cutaneous reflex. AB - Our previous studies have shown that the cerebral cortex plays a modulator role in the physiological mechanisms of acupuncture and 2Hz electroacupuncture (EA), but either acupuncture or 2Hz EA appeared to have very little effect on spinal cord. However, 2Hz or 100Hz transcutaneous electrical nerve stimulation (TENS) applied to Hegu acupuncture point (LI.4) can increase the amplitude of the H reflex, whereas manual acupuncture has no similar effect. Therefore, the aim of the present study was to further investigated the effect of 2Hz EA, 2Hz or 100Hz TENS on cutaneous reflex (CR). A total of 13 healthy adult volunteers were studied. CR recordings were obtained on the right first dorsal interosseous muscle by electrical stimulation of the ipsilateral index finger. 2Hz EA, 2Hz or 100Hz TENS was applied to the surface of the left LI.4, and then the changes of CR were observed. The results indicated that CR was characterized by an initial short latency excitatory wave (E1), followed by an inhibitory wave (I1), then a long latency excitatory wave (E2). Both 2Hz EA and 2Hz TENS applied to the surface of the left LI.4 could prolong the latencies of I1 component of CR, whereas 100Hz TENS has no similar effect. In addition, either EA or TENS could not change the latencies of E1, E2 component and the amplitudes of I1, E2 component of CR. In conclusion, 2Hz EA or 2Hz TENS applied to the LI.4 prolonged the latencies of I1 component of CR, suggesting that the action site of 2Hz EA or 2Hz TENS located in the supraspinal, possibly in the subcortical or cortical level. In addition, the electrical stimulation of higher frequency such as 100Hz TENS acting on CR caused habituation easier. PMID- 12638740 TI - The First Baltic Symposium on Environmental Chemistry. 26-29 September 2001, Tartu, Estonia. PMID- 12638741 TI - The change in O3, SO2 and NO2 concentrations in Lithuania. AB - Due to the dynamic nature of the atmosphere, substantial amounts of gaseous and particulate pollutants are transported to the areas distant from their sources. In order to determine the regional concentration levels of atmospheric pollutants in Lithuania, concentrations of gaseous O3, SO2, NO2 and other pollutants have been measured at the Preila background station (55 degrees 20' N and 21 degrees 00' E, 5 m a.s.l.) since 1981. The long-term concentration data set enabled us to get temporal trends, both on a seasonal and longer time scale, to identify source areas of pollutants and to relate them to the emission data. Based on the data obtained, the different tendencies in the pollutant concentration changes were revealed. Positive trends for ozone (of 2.9% per year during 1983-2000) and a distinct negative trend for both sulphur dioxide (of 3.8% per year during 1981 2000) and nitrogen dioxide (of 3.8% per year during 1983-2000) were found. The air mass back-trajectory analysis was used to assess the source region of air pollutants transported to Lithuania. The pollutant concentration levels were compared with their emission changes in Europe and Lithuania. The general trends in SO2 as well as in NO2 concentrations observed are consistent with changes in SO2 and NO2 emissions in Europe and Lithuania. PMID- 12638742 TI - Analysis of river pollution data from low-flow period by means of multivariate techniques: a case study from the oil-shale industry region, northeastern Estonia. AB - The oil-shale industry has created serious pollution problems in northeastern Estonia. Untreated, phenol-rich leachate from semi-coke mounds formed as a by product of oil-shale processing is discharged into the Baltic Sea via channels and rivers. An exploratory analysis of water chemical and microbiological data sets from the low-flow period was carried out using different multivariate analysis techniques. Principal component analysis allowed us to distinguish different locations in the river system. The riverine microbial community response to water chemical parameters was assessed by co-inertia analysis. Water pH, COD and total nitrogen were negatively related to the number of biodegradative bacteria, while oxygen concentration promoted the abundance of these bacteria. The results demonstrate the utility of multivariate statistical techniques as tools for estimating the magnitude and extent of pollution based on river water chemical and microbiological parameters. An evaluation of river chemical and microbiological data suggests that the ambient natural attenuation mechanisms only partly eliminate pollutants from river water, and that a sufficient reduction of more recalcitrant compounds could be achieved through the reduction of wastewater discharge from the oil-shale chemical industry into the rivers. PMID- 12638743 TI - Biomarkers of carcinogenic contaminants in Baltic flounder (Platichthys flesus): temporal changes in urban and non-urban sites in Tallinn bay. AB - In order to study effects of environmental contamination, a suite of biomarkers were measured over the period 1996 to 1999 in livers of flounder (Platichthys flesus) from two urban embayments and one non-urban reference site of the Gulf of Finland in the vicinity of Tallinn, Estonia. Total cytochrome P450 (CYP) level, aryl hydrocarbon hydroxylase (AHH), 5-aminolevulinic acid synthetase (ALA-S), and heme synthetase (HEM-S) activities were quantified by means of spectrophotometry. These data were compared to results obtained in 1994 for the same biomarkers at one of the urban embayments and the non-urban site, as measured by the same protocols. For the flounder collected from the non-urban site, changes occurred in AHH activity and the total CYP level, which were significantly lower in 1996 and 1999 compared with 1994 (p < 0.05). Activity of ALA-S decreased slightly over this same period. The activity of HEM-S increased between 1996 and 1999. In the urban site first investigated in 1994, the activities of AHH and ALA-S, as well as the total level of CYP in flounder liver were significantly higher compared with 1999 (p < 0.05). HEM-S activities did not show any significant changes over this time period. AHH activities of flounder collected in another urban site decreased slightly between 1996 and 1999, in contrast to data on the total CYP level which diminished drastically over these years (p < 0.05). Activities of HEM S increased significantly (p < 0.05) during the period investigated, while activities of ALA-S remained unchanged. These findings suggest that contamination of the marine environments by PAHs has gone down everywhere in the Tallinn area during the last 3 to 5 years. However, the results indicate that the area is still contaminated, as indicated by elevated heme synthesis enzymes and the total CYP content, and that monitoring of contaminants and their effects should be continued in this region. PMID- 12638744 TI - Biodegradation of dimethylphenols by bacteria with different ring-cleavage pathways of phenolic compounds. AB - The biodegradation of 3,4, 2,4, 2,3, 2,6 and 3,5-dimethylphenol in combination with phenol and p-cresol by axenic and mixed cultures of bacteria was investigated. The strains, which degrade phenol and p-cresol through different catabolic pathways, were isolated from river water continuously polluted with phenolic compounds of leachate of oil shale semicoke ash heaps. The proper research of degradation of 2,4 and 3,4-dimethylphenol in multinutrient environments was performed. The degradation of phenolic compounds from mixtures indicated a flux of substrates into different catabolic pathways. Catechol 2,3 dioxygenase activity was induced by dimethylphenols in Pseudomonas mendocina PC1, where meta cleavage pathway was functional during the degradation of p-cresol. In the case of strains PC18 and PC24 of P. fluorescens, the degradation of p-cresol occurred via the protocatechuate ortho pathway and the key enzyme of this pathway, p-cresol methylhydroxylase, was also induced by dimethylphenols. 2,4 and 3,4-dimethylphenols were converted into the dead-end products 4-hydroxy-3 methylbenzoic acid and 4-hydroxy-2-methylbenzoic acid. In the degradation of 3,4 dimethylphenol, the transient accumulation of 4-hydroxy-2-methylbenzaldehyde repressed the consumption of phenol from substrate mixtures. A mixed culture of strains with different catabolic types made it possible to overcome the incompatibilities at degradation of studied substrate mixtures. PMID- 12638745 TI - The toxicity and fate of phenolic pollutants in the contaminated soils associated with the oil-shale industry. AB - Phenol, cresols, dimethylphenols and resorcinols considered major pollutants in the oil-shale semi-coke dump leachates (up to 380 mg phenols/L) that contaminate the surrounding soils and pose a threat to the groundwater in the North-East of Estonia. However; despite high residual concentrations of polyaromatic hydrocarbons (PAHs) and oil products in these soils, the concentration of phenols (especially their water-extractable fraction) was low, not exceeding 0.7 mg/kg dwt. The aim of the current study was to evaluate the role of biodegradation and aging on the decrease of hazard caused by phenolic pollution. The extractability of phenols (phenol, cresols, dimethylphenols and resorcinols) and their biodegradability by the microbial population was studied in the 13 soils sampled from the Estonian oil-shale region, territories of former gas stations, and from presumably non-polluted areas. Phenol, 5-methylresorcinol, p-cresol and resorcinol could be considered easily degradable in the soils as the microbial populations from majority of the soils studied were able to grow on mineral medium supplemented with these phenols as a single source of carbon. 2,3- and 2,4 and 3,4-dimethylphenols could be considered less easily biodegradable. The semi coke dump leachate polluted soil (containing no dibasic phenols, 43 mg of monobasic phenols, 1348 mg of oil products and 35 mg of PAHs per g dwt) was analyzed chemically (HPLC) and toxicologically (Flash-Assay using Vibrio fischeri) for the leaching of phenols during shaking of soil-water slurries for 24 h. Only 5.8% of the total concentration of phenols was water-extractable, whereas about 50% of the leached amount was biodegraded by the soil microorganisms. Phenol and cresols were biodegraded by 80%, but the concentration of dimethylphenols practically did not change. The pollutants (measured as total water-extractable toxicity) were desorbed from the soil particles by the 8th h of extraction, whereas the toxicity of the aqueous phase continued to increase, probably due to the formation of toxic metabolites. The concentration of water extractable phenols was too low to explain the toxicity of the extract. Also the impact of PAHs and oil products was excluded. Thus, the relatively low concentration of phenols in the oil-shale region soils is most probably the reflection of both natural attenuation and pollution aging. Therefore, the impact of phenolic compounds to the net bioavailable hazard is probably not so remarkable as has been considered. The actual pollutants causing the soils from the oil-shale region, however, need to be elucidated. PMID- 12638746 TI - Applied waste-free recovery of methanol: a sustainable solution for chromatography laboratories. AB - In this paper, we present appliedmethanol recycling technology utilising chromatographic applications, which has been designed for an academic-size institution. The procedure is combined out of proper recovery technique and the biodegradation method intended for postprocessing residues. Additionally, analytical methods controlling the quality of the process are described in detail in order to enable full transfer of the proposed methodology to the analogous institution. The recovered solvent is of relatively high purity (> 99.92%), containing traces of water and volatile compounds. The spectral purity is sufficient to employ recycled methanol in HPLC applications where low wavelength detection is necessary. Biodegradation of distillation first-cuts and still bottoms is implemented using isolated strains of Methylobacterium organophilum. During the biodegradation process, a series of carotenoids are biosynthesized, which are of a high commercial value. The proposed application, therefore, offers not only a sustainable, waste-free solution in handling methanol waste, but can also serve as a potential source of value. PMID- 12638747 TI - The effect of surfactants on oxygen mass-transfer through the air-water interface. AB - The aim of the current study was to investigate the oxygen transfer rate through the water-air interface in the presence of different concentrations of various surfactants. The surfactants used in the current study were methanol, ethanol, 1 propanol and 1-butanol. The measurements were performed with an apparatus based on the electrochemical oxygen sensor. Theresults obtained showed that the impediment to oxygen mass-transfer through the water-air interface depends on the molecular structure and concentration of the surfactant used. A simple mathematical model was proposed, which allowed us to calculate the characteristic constants of surfactants--K' and deltak starting from the experimental results. K' is the reciprocal of the equilibrium constant of adsorption process and deltak characterizes the influence of the surfactant to oxygen mass-transfer through the air-water interface. PMID- 12638748 TI - The Langmuir two-surface equation as a model for cadmium adsorption on peat: a new method of the calculation of equation constants. AB - The Langmuir isotherm equation is often used to describe the adsorption of cadmium on peat. If the total surface consists of two or more populations of sites with different bonding energies, it is proposed that the Langmuir two surface equation be used. The effects of the contact time and pH on adsorption were investigated in batch experiments. Adsorption batch isotherm studies were carried out by using 12 solutions with initial concentrations of 5-110 mg/l. The samples were analysed by using axial ICP-AES. Adsorption data were compared using four different linearizations (Lineweaver-Burk, Eadie-Hofstee, Scatchard and Langmuir linearizations). Better results among linearizations were found with the Langmuir linearization, but the results obtained by using the nonlinear regression were still the best. A new, nonlinear regression method for the calculation of the equation constants was created by using the system of equations and these findings were compared to the results found using earlier methods. PMID- 12638749 TI - Remediation of an old landsfill site: soil analysis, leachate quality and gas production. AB - The Baltic Sea Region has a large number of landfills that need remediation after care routines and control, in order to avoid future emissions to the environment and to fulfil the demands in the EU Waste Council Directive on the landfill of waste. Based on the Masalycke test screening, an excavation of whole or of parts of the landfill can be seen as a potential measure for some of the old landfills. The material excavated in the test was screened into the fractions: < 18 mm, 18 50 mm and > 50 mm. The coarsest fraction (> 50 mm) contained 50% wood and paper. The medium-sized fraction (18-50 mm) contained stones and indefinable soil-like material, while the fine fraction contained peat-like material with some other small waste components. The spectral analysis of heavy metals indicated only high concentrations of zinc and there was no significant difference between the fine and the medium-sized fractions. The medium sized and the unsorted fraction was moisturized and refilled into the pit. The methane content in the landfill gas from the pit was 50-57% in the sorted material with a flow of 8-17 l/min and 38 57% in the unsorted fraction with a flow of 2-13 l/min during the first 1.5 year. The Masalycke landfill is in the methanogenic phase and leachate concentrations are normal. Landfill mining can be used to prolong the landfill lifetime and/or used as a tool for remedial actions in contaminated sites. PMID- 12638750 TI - Application of new physical chemical methods in soil ecological investigations. AB - The article discusses methodological investigations for the improvement and unification of soil testing in combination with the application of complex physico-chemical methods. An analytical procedure involving different extractions was used in order to determine the total and mobile amount of heavy metals (Cd, Cr, Pb, Ni, Cu, Zn, etc.) by atomic absorption spectrophotometry in soil and its fine-dispersive fraction (< 0.005 mm). The average samples (effected upon by background pollution) of Calcari Epihypogleyic Luvisol, (Lvg-p-w-cc, FAO-Unesco, 1998) has been taken from the rotation field of the experimental station of the Lithuanian University of Agriculture Subsequently, a fine-dispersive fraction was separated by a principle of peptization in distilled water. The investigation results obtained have shown a substantial dependence on the extractor used and the amount of fine-dispersive fraction in soil as well. It was found that the greatest reliability of the mobile heavy metal form is by using 1N CH3 COONH4 extractor and an HCl+HF mixture extractor for their total amount. Additionally, for the first time in Lithuania, the X-ray photoelectron spectroscopy (XPS) it has been applied for the interpretation of soil chemical composition. PMID- 12638751 TI - Knudsen cell construction, validation and studies of the uptake of oxygenated fuel additives on soot. AB - INTENTION, GOAL, SCOPE, BACKGROUND: The properties of atmospheric particles are important to public health, radiative forcing of the atmosphere and to elucidating the chemical reactivity of atmospheric particles. We have constructed a Knudsen cell to study the uptake of organic compounds on soot. This article describes the construction and validation of the instrument, and our results on commercial soot concerning the uptake coefficient of ethanol, acetone, 1-butanol and diethoxymethane. OBJECTIVES: First, a technical description of the instrument is presented. Next, its performance is validated by measuring the uptake of NO2 on hexane soot. Finally, the uptake coefficients of four oxygenated hydrocarbons on commercial soot are presented. The objective is to contribute to the understanding of the formation of particles in motor vehicle exhaust. METHODS: A Knudsen cell is used to measure the uptake of specific gas-surface systems. A quadrupole mass spectrometer is used to determine the decay rate of a pulse of reagent gas in the reaction chamber. RESULTS AND DISCUSSION: The BET surface area of the commercial soot was 12.6 m2/g. The uptake coefficient (gamma) has been determined for ethanol (gamma0,BET = 7.7 +/- 4.8 x 10(-8)), 1-butanol 1.4 +/- 0.54 x 10(-7)), acetone (gamma0BET = 1.5 +/- 0.15 x 10(-7)) and diethoxymethane (gamma0,BET = 2.6 +/- 0.61 x 10(-7). These results are characteristic of the specific soot sample used. The ordering of the uptake coefficients, ethanol < 1 butanol approximately acetone < diethoxymethane, can be ascribed to a combination of physical (size and mass) and chemical effects. In addition, the initial uptake coefficient for NO2 on fresh hexane soot was determined to be gamma0,BET = 1.7 +/ 1.1 x 10(-4). CONCLUSIONS: In conclusion, we demonstrate that this instrument is able to measure uptake coefficients that are in agreement with accepted literature values. New data is presented concerning four light oxygenated hydrocarbons. RECOMMENDATIONS AND OUTLOOK: A large amount of detailed information concerning individual heterogeneous reactions is necessary in order to model the composition of motor vehicle emissions. We look forward to increasing the size of this database. Results for a series of alcohols and alkanes will be presented in a forthcoming publication. PMID- 12638753 TI - Nitrogen fluxes in the Tyrolean Limestone Alps. PMID- 12638752 TI - The comparison study of compost and natural organic matter samples. AB - Sewage sludge is a serious problem facing modern wastewater treatment plants. One of the methods to safely utilize the sludge is through composting and the agricultural use of the final product. In this study the compost samples from municipal sewage sludge are compared to the natural, rich in organic matter samples. Based on physicochemical properties of the material, the maturity indices are obtained to evaluate the quality of organic matter and estimate the similarities between the samples. The study shows that one-dimensional analysis like that based on maturity indices does not provide satisfactory answers concerning the nature of such complex materials. The data set of the analysis naturally possesses a multidimensional character so that the employment of advanced chemometric techniques like cluster analysis show a number of features which were hidden within the 'data flood'. The major conclusion of this work is that the compost from sewage sludge is similar to peat in the significance of the properties of the organic matter. Moreover, the organic matter of agricultural soil, which is most stable and has been naturally matured over a long period of time, differs substantially from the other samples. PMID- 12638754 TI - Nitrogen fluxes on an intensive investigation plot in the North Tyrolean Limestone Alps. AB - In the framework of this study, nitrogen fluxes on a limestone site are investigated. The major goals are the assessment of the nitrogen status, the estimation of the nitrogen budget and the evaluation of the nitrogen saturation. The investigation area, the intensive investigation plot and the research equipment are described. PMID- 12638755 TI - Analyses of NOx and wet depositions at Muhleggerkbopfl, North Tyrolean Limestone Alps. AB - The intensive investigation site 'Muhleggerkopfl' in the North Tyrolean Limestone Alps can be classified as a clean-air area site. The mean concentrations of NOx are far below the effect-related limit value of the WHO (30 microg NOx m(-3)). The gravitational depositions in the open field (bulk deposition) ranged from 10.8 to 14.7 kg N ha(-1) a(-1) (throughfall: 11.3 to 12.3 kg N ha(-1) a(-1)) in the measuring years 1998 to 2000. Compared to these data, depositions in other forested areas of the Austrian Alps amounted to up to 30 kg N ha(-1) a(-1). The gravitational depositions (bulk deposition) alone--without considering dry and occult deposition slightly exceeded the lower limit of Critical Loads for coniferous and deciduous forests (>10 kg N ha(-1) a(-1)), but were below the Critical Loads for calcareous forests (15-20 kg N ha(-1) a(-1)). PMID- 12638757 TI - Gaseous nitrogen losses from a forest site in the North Tyrolean Limestone Alps. AB - Microorganisms are responsible for the mineralisation of organic nitrogen in soils. NH4+ can be further oxidised to NO3- during nitrification and NO3- can be reduced to gaseous nitrogen compounds during denitrification. During both processes, nitrous oxide (N2O), which is known as greenhouse gas, can be lost from the ecosystem. The aim of this study was to quantify N2O emissions and the internal microbial N cycle including net N mineralisation and net nitrification in a montane forest ecosystem in the North Tyrolean Limestone Alps during an 18 month measurement period and to estimate the importance of these fluxes in comparison with other components of the N cycle. Gas samples were taken every 2 weeks using the closed chamber method. Additionally, CO2 emission rates were measured to estimate soil respiration activity. Net mineralisation and net nitrification rates were determined by the buried bag method every month. Ion exchange resin bags were used to determine the N availability in the root zone. Mean N2O emission rate was 0.9 kg N ha(-1) a(-1), which corresponds to 5% of the N deposited in the forest ecosystem. The main influencing factors were air and soil temperature and NO3- accumulated on the ion exchange resin bags. In the course of net ammonification, 14 kg NH4+-N ha(-1) were produced per year. About the same amount of NO3--N was formed during nitrification, indicating a rather complete nitrification going on at the site. NO3- concentrations found on the ion exchange resin bags were about 3 times as high as NO3- produced during net nitrification, indicating substantial NO3- immobilisation. The results of this study indicate significant nitrification activities taking place at the Muhleggerkopfl. PMID- 12638756 TI - Altitude-dependent wet, dry and occult nitrogen deposition in an Alpine region (Achenldrch, Austria, 920 m - 1758 m a.s.l.). AB - From November 1995 to October 1996 and from October 1997 to September 1998, samples of wet precipitation, cloud water as well as of reactive gases and particulate matter, were collected at three elevational levels (920 m, 1,280 m and 1,758 m a.s.l.) in Achenkirch, Austria. The samples were analysed for ammonium and nitrate in wet precipitation and in cloud water, for ammonia, nitric acid and nitrogen dioxide in the gas phase and for particulate ammonium and particulate nitrate in aerosol. Total nitrogen deposition was calculated combining measured concentrations in wet, dry and occult depositions with the corresponding deposition fluxes. Two multilayer deposition models were used for the calculation of dry and occult deposition. The total nitrogen input in 1995/96 was estimated to be 29 kg N ha(-1)a(-1) at the Christlumkopf station (1,758 m), 20 kg N ha(-1)a(-1) at the Christlumalm station (1,280 m) and 28 kg N ha(-1)a(-1) at the Talboden station (930 m). Respective data for the 1997/98 observation period were 31 kg N ha(-1)a(-1) at the Christlumkopf station (1,758 m) and 18 kg N ha(-1)a(-1) at the Mulhleggerkopfl station (920 m). Critical Loads of nitrogen for coniferous forests were exceeded significantly near-source regions represented by areas of intense agricultural use and at high elevation sites. PMID- 12638758 TI - Water and nitrate fluxes at a forest site in the North Tyrolean Limestone Alps. AB - The water balance for the site Muhleggerkopfl in the North Tyrolean Limestone Alps has been established to a soil depth of 50 cm. The evaporation amounts to 42% and deep percolation is 58% of the precipitation. The surface runoff was negligible and therefore the according nitrate fluxes as welL Soil water analysis revealed mean nitrate concentrations of 3 to 15 mg NO3 L(-1), depending on soil depth. The nitrate concentrations at 50 cm soil depth and the associated percolation rates led to NO3-N outputs of 15.9 kg NO3-N ha(-1) in the year 1999 and 7.9 kg NO3-N ha(-1) in the year 2000. PMID- 12638759 TI - Nitrate dynamics in an Alpine forest site (Muhleggerkopfl). O and N stable isotope analysis in natural water samples. AB - Stable isotope analysis of 15N/14N and 18O/16O-nitrate was used to investigate the nitrate dynamics and potential groundwater pollution in an Alpine forest stand in Tyrol/Austria. The dynamics of delta15-N(nitrate) values were followed in a forest ecosystem. The stable isotopic values of the throughfall are comparable with other studies. The completely decoupled dynamics of the delta15 N(nitrate) of the precipitation and the surface water was observed. High variations in delta15N-nitrate values in rainfall indicate that nitrate of different sources is deposited at that site. A significant correlation between the delta15N(nitrate) values of the surface water and soil water was obtained, while no significant correlation between the delta15N(nitrate) values of any precipitation sample with the surface water could be found. This suggests that the main source of nitrate in soil water originates from microbiological activity such as nitrification reactions and less from nitrate input by deposition. The results of delta18O(nitrate) measurements strongly supported the microbiological origin of nitrate in the surface and soil water In an additional lysimeter experiment, 15N-labelled nitrate was applied to study nitrate transport in soil. After 130 days and the collection of 300 L leachate, a total of 52% of the applied nitrate was detected in seepage water. PMID- 12638760 TI - Simulation of soil hydrology and establishment of a nitrogen budget of a mountain forest. AB - The water fluxes through the mountainous forest ecosystem 'Muhleggerkopfl' were simulated by means of the mechanistic soil physical model Hydrus 1D. The objective was to set up a nitrogen budget in order to decide if the ecosystem accumulates nitrogen or if nitrogen leaks from the site. The simulated annual loss of N by percolation ranges between 0.4 and 1 g N m(-2) yr(-1) and is smaller than the annual input by bulk and occult deposition, which combines to approx 1.2 1.5 g N m(-2) yr(-1). Obviously the forest soil presently accumulates N. With an N input-rate exceeding the N output, the operationally defined status of N saturation is not yet reached. Comparing the magnitude of the N pool in the soil (several kg N m(-2)) with the rate of the annual increase (a few g N m(-2) yr( 1)), the process of N saturation is apparently slow. PMID- 12638761 TI - Investigations of nitrogen fluxes and pools on a limestone site in the Alps. AB - In the North Tyrolean Limestone Alps a site was investigated over a four-year period (1998-2001) in order to assess the nitrogen saturation status, the nitrogen budget (quantification of the net uptake of nitrogen by the canopy and of the nitrogen mineralization, nitrogen uptake from roots and N2O emission rates, proof of the origin of nitrate in the soil water with stable isotope analyses), and the effects of the actual nitrogen input on ground water quality. The main goals were to quantify the nitrogen input rate, the nitrogen pools in above-ground and below-ground compartments, nitrogen turnover processes in the soil as well as the output into the groundwater and into the atmosphere. The findings are based on continuous and discontinuous field measurements as well as on model results. While nitrogen input exceeded the Critical Loads of the WHO (1995), nitrogen deficiency and nutrient imbalances were verified by needle analyses. The atmospheric input of inorganic nitrogen was higher than the nitrogen output in 50 cm soil depth. A tracer experiment with 15N helped to prove that not more than half of the applied nitrate could be discharged. This allows the conclusion that nitrogen is stored in the system and that the site cannot yet be said to be saturated with nitrogen. The same result was also obtained by modelling. In addition, it was proved that the nitrogen discharge did not stem from deposition but from processes within the system. PMID- 12638762 TI - The Parent Interview for Autism-Clinical Version (PIA-CV): a measure of behavioral change for young children with autism. AB - The Parent Interview for Autism-Clinical Version (PIA-CV) was developed to measure autism symptom severity across a wide range of behavioral domains. Two studies were conducted to examine the psychometric properties of the PIA-CV for a sample of children under 3 years old. Results of study 1 revealed adequate internal consistency for nine of the 11 PIA-CV dimensions, as well as significant group differences on social-communication domains between 2-year-old children with autism and a developmentally matched sample. Study 2 examined the association between changes in PIA-CV scores and changes in autism symptomatology from age 2 to age 4. Results revealed that changes on PIA-CV dimensions assessing social and communication skills were associated with clinically significant behavioral and diagnostic improvements. These findings support the utility of the PIA-CV for obtaining ecologically valid information from parents and for measuring behavioral change in young children with autism. PMID- 12638763 TI - The disclosure of a diagnosis of an autistic spectrum disorder: determinants of satisfaction in a sample of Scottish parents. AB - Satisfaction with disclosure of the diagnosis of an autistic spectrum disorder was investigated using a self-report questionnaire completed by 126 parents. On a rating of satisfaction, 55 percent indicated that they were satisfied or very satisfied with the disclosure. Parents were more likely to be satisfied if they gave positive ratings to the manner of the professional and the quality of the information provided; if they had been given written information and the opportunity to ask questions; and if their early suspicions had been accepted by professionals. These factors were combined into a global index of satisfaction; those gaining higher scores were more likely to have been given the diagnosis of Asperger syndrome (as opposed to autism), to have had a definite diagnosis, and to have children who were not currently in an educational placement. These results underline the importance of the interaction between parent and professional during the disclosure interview. PMID- 12638764 TI - How well does early diagnosis of autism stand the test of time? Follow-up study of children assessed for autism at age 2 and development of an early diagnostic service. AB - Twenty children who presented with severe and communication difficulties at age 2 underwent a comprehensive assessment for autism, and were reassessed at age 4-5. In common with other recent studies, diagnosis of autistic spectrum disorders at age 2 was found to be reliable and stable. The communication and social skills of the children showed little change overall by the second assessment. However, children whose scores deteriorated in the social domain tended to have presented initially with more significant behaviour problems. Few repetitive behaviours were observed at age 2, whereas these were more apparent by age 4-5. The finding that early diagnosis of autism is reliable and stable has led to the development of an early diagnostic service in Southampton, which is described. The importance of early diagnosis is that it opens the door to early intervention programmes, which in turn prevent many problems from occurring in later life. PMID- 12638765 TI - Ability profiles in children with autism: influence of age and IQ. AB - To understand the effect of IQ and age on ability in children with autism, psychological data were analyzed for 164 3- to 15-year-olds with autism (IQs 14 143). As age increased, so did IQ, which probably reflects both an actual increase in IQ over time and the likelihood that brighter children are diagnosed later. Early in life, 67 percent had normal motor and delayed speech milestones. Verbal IQ continued to lag behind non-verbal IQ during the preschool years. By school age, the gap between verbal and non-verbal IQs had closed. Visual reasoning exceeded graphomotor scores for all children, and surpassed IQ for most. Graphomotor scores were significantly below IQ for both high-IQ groups. For school-age children with low IQs, math, spelling, and writing scores were consistent with IQ and reading was above IQ. School-age children with high IQs had average reading, math, and spelling scores and a weakness in writing. PMID- 12638766 TI - The development and maintenance of friendship in high-functioning children with autism: maternal perceptions. AB - The current study investigated mothers' perceptions of the development of friendship in high-functioning children with autism and in typically developing children. Fourteen mothers in each group (autism, typical) completed the Childhood Friendship Survey regarding their children's friendships. Main results indicated that both groups (autism and typical) tended to have same-gender and same-age friendships. However, friendships of children with autism differ compared with typical children's friendships on number of friends, friendship duration, frequency of meetings, and type of activities. Half of the friendships in the autism group were mixed (friendship with a typically developing child). Mixed differed from non-mixed friendships in that mixed pairs met and played mostly at home, whereas non-mixed pairs met and played at school. Factors contributing to the development and formation of friendship in each group are discussed. PMID- 12638767 TI - Psychosocial functioning in a group of Swedish adults with Asperger syndrome or high-functioning autism. AB - This study reports on psychosocial functioning in Swedish adults with Asperger syndrome (AS) or high-functioning autism (HFA). A systematically selected sample of patients and relatives was interviewed concerning their psychosocial situation. The majority was living independently. All persons but one were unemployed. None was married and none had children. Only a few had some kind of partner. Most persons needed a high level of public and/or private support. The overall adjustment was rated good in 12 percent, fair in 75 percent and poor in 12 percent. Adult persons with AS/HFA have extensive need for support from their families and/or society. This information is important in order to provide adequate interventions that are in accordance with the expressed needs of the individuals themselves. PMID- 12638768 TI - Esthetic treatment of enlarged gingival embrasures. PMID- 12638769 TI - The ZiReal Post: A new ceramic implant abutment. AB - Restorations in the anterior esthetic zone present significant challenges in both the surgical and prosthetic phases of implant dentistry. Titanium has been established as the material of choice for endosseous implants, resulting in a high degree of predictability. Many types of implants require transmucosal abutments to retain implant restorations. Ceramics may be the ideal material to replace natural teeth, but most transmucosal abutments are made of titanium. However, ceramics may also be used as abutments in implant restorations. This combination of ceramics for abutment and crown provides better translucency for the implant restoration than is available with metal abutments and porcelain fused-to-metal crowns. Ceramic abutments and implant restorations also minimize the gray color associated with metal components that is transmitted through the peri-implant tissues. Customized emergence profiles also may be obtained with ceramic abutments; this generally improves the predictability and consistency of the esthetics obtainable in implant restorations. Zirconia as a ceramic material offers not only outstanding material properties but also a well-documented biocompatibility. PMID- 12638770 TI - Fracture resistance of endodontically treated teeth restored with three different prefabricated esthetic posts. AB - PURPOSE: This study was undertaken to evaluate the role of composition of prefabricated esthetic posts in fracture resistance of endodontically treated teeth in vitro. MATERIALS AND METHODS: Thirty human, single-rooted teeth (maxillary central incisors and canines) with similar root dimensions, extracted for therapeutic reasons, were used in this study. The crowns were removed below the cementoenamel junction to obtain a standard root length of 17 mm. The roots were endodontically treated following the conventional manual technique and randomly assigned to three groups (n = 10) according to the post used: Aestheti Post, Bisco, Schaumberg, Illinois; FibreKor Post, Jeneric/Pentron, Wallingford, Connecticut; and CosmoPost, Ivoclar Vivadent, Schaan, Liechtenstein. The root canals were rendered patent, and the root preparations were standardized through flaring with the manufacturers' drills included in the respective kits. The posts were cemented with an adhesive system and a resin cement (All-Bond 2 and C&B, Bisco, respectively), according to the manufacturers' directions. Composite resin (Tetric Ceram, Ivoclar Vivadent) crowns were built up using a preformed polyester matrix, and the specimens were mounted in metallic rings with cold-cure acrylic resin and kept in saline solution at 4 degrees C for 24 hours. Fracture resistance was then determined using an EMIC DL-2000 universal testing machine. The crosshead speed was 0.5 mm/min with the 45-degree compressing load at the middle third of the crown. Data were analyzed using one-way analysis of variance and Tukey's test (p < or = .001). RESULTS: Mean fracture resistance was as follows: Aestheti-Post, 83.5 kgf; FibreKor Post, 85.7 kgf; and CosmoPost, 36.5 kgf. The fracture strength of CosmoPost was significantly lower than that of the other posts. Teeth restored with CosmoPost had post fractures, and in three specimens, those were associated with root fractures. Teeth restored with the other two posts presented fractures on the composite crowns. PMID- 12638771 TI - Bonding characteristics of self-etching adhesives to intact versus prepared enamel. AB - PURPOSE: This study tested the null hypothesis that the preparation of the enamel surface would not affect the enamel microtensile bond strengths of self-etching adhesive materials. MATERIALS AND METHODS: Ten bovine incisors were trimmed with a diamond saw to obtain a squared enamel surface with an area of 8 x 8 mm. The specimens were randomly assigned to five adhesives: (1) ABF (Kuraray), an experimental two-bottle self-etching adhesive; (2) Clearfil SE Bond (Kuraray), a two-bottle self-etching adhesive; (3) One-Up Bond F (Tokuyama), an all-in-one adhesive; (4) Prompt L-Pop (3M ESPE), an all-in-one adhesive; and (5) Single Bond (3M ESPE), a two-bottle total-etch adhesive used as positive control. For each specimen, one half was roughened with a diamond bur for 5 seconds under water spray, whereas the other half was left unprepared. The adhesives were applied as per manufacturers' directions. A universal hybrid composite resin (Filtek Z250, 3M ESPE) was inserted in three layers of 1.5 mm each and light-cured. Specimens were sectioned in X and Y directions to obtain bonded sticks with a cross sectional area of 0.8 +/- 0.2 mm2. Sticks were tested in tension in an Instron at a cross-speed of 1 mm per minute. Statistical analysis was carried out with two way analysis of variance and Duncan's test at p < .05. Ten extra specimens were processed for observation under a field-emission scanning electron microscope. RESULTS: Single Bond, the total-etch adhesive, resulted in statistically higher microtensile bond strength than any of the other adhesives regardless of the enamel preparation (unprepared = 31.5 MPa; prepared = 34.9 MPa, not statistically different at p < .05). All the self-etching adhesives resulted in higher microtensile bond strength when enamel was roughened than when enamel was left unprepared. However, for ABF and for Clearfil SE Bond this difference was not statistically significant at p > .05. When applied to ground enamel, mean bond strengths of Prompt L-Pop were not statistically different from those of Clearfil SE Bond and ABF. One-Up Bond F did not bond to unprepared enamel. Commercial self etching adhesives performed better on prepared enamel than on unprepared enamel. The field-emission scanning electron microscope revealed a deep interprismatic etching pattern for the total-etch adhesive, whereas the self-etching systems resulted in an etching pattern ranging from absent to moderate. PMID- 12638772 TI - Six-month storage-time evaluation of one-bottle adhesive systems to dentin. AB - PURPOSE: The goal of this study was to evaluate the 1-week, 3-month, and 6-month performance of eight commercially available one-bottle adhesive systems to dentin. MATERIALS AND METHODS: Lingual and buccal surfaces from human third molars were ground wet on 600-grit SiC paper to obtain a flat dentinal surface. The specimens were randomly divided into 24 groups (n = 10), which were established to measure the shear bond strengths of Bond-1 (B1), ONE-STEP (OS), OptiBond SOLO (OP), Prime & Bond 2.1 (PB), Single Bond (SB), STAE (ST), Syntac Sprint (SS), and Tenure Quick (TQ) after 1-week, 3-month, and 6-month water storage at 37 degrees C. One-bottle adhesives were applied according to manufacturers' instructions and Z100 composite cylinders were applied on the bonded dentinal surfaces. The 3-month water-storage groups were thermocycled for 1500 cycles at 5 degrees C and 55 degrees C and 6-month groups for 3000 cycles. After storage periods, specimens were tested in shear in a universal testing machine (0.5 mm/min). RESULTS: were statistically analyzed by analysis of variance and Tukey test. Results: The changes in shear bond strengths were not uniform over time. Over the test period, OS, PB, SB, and SS exhibited bond strength stability, however, SS presented low bond strengths on all tested periods. A significant decrease in bond strength was observed for B1, OP, ST, and TQ after the 6-month storage period. PMID- 12638774 TI - New materials: risk or opportunity? PMID- 12638773 TI - Clinical evaluation of a 3% hydrogen peroxide tooth-whitening gel. AB - PURPOSE: The purpose of this randomized double-blind investigation was to clinically evaluate the effectiveness of a commercially available 3% hydrogen peroxide tooth-whitening material (ADS Tooth Whitening Gel, Applied Dental Sciences, Inc., Lee, MA). The material was used over 2 weeks in a dentist supervised, patient-applied application. MATERIALS AND METHODS: The evaluation was carried out in accordance with American Dental Association (ADA) Council on Scientific Affairs Acceptance Program Guidelines for Home-Use Tooth Whitening Products (May 1998). Sixty-five participants were randomized into two groups (active and placebo) for a double-blind, placebo-controlled study. A custom application tray, scalloped without reservoirs, was fabricated for each participant. Tooth shades were documented by matching with the Vita shade guide ordered according to value and assigned a numeric ranking from 1 to 16, darkest to lightest. Participants were supplied with the treatment solutions to use in the application tray. Duration of active treatment was 30 minutes 3 times a day for 2 weeks. Participants were evaluated at baseline, 1 week and 2 weeks treatment, and 12 and 26 weeks post-treatment. The statistical analysis used at each time point to determine the difference between the active and placebo groups based on the change in value was the two-sample t-test. Plaque and soft tissue indices were also recorded. RESULTS: The teeth treated with 3% hydrogen peroxide were significantly lighter at 2, 12, and 26 weeks (p < or = .0140, .0004, and .0001, respectively) compared with the placebo group. The mean shade change was 4.2 Vita shade tabs at 2 weeks. At 26 weeks (6 months), the degree of whitening was 4.1 tabs. PMID- 12638775 TI - Longevity of ceramic inlays/onlays: Part II. PMID- 12638776 TI - Mechanical toothbrush. PMID- 12638777 TI - Androgen receptor gene and male infertility. AB - Androgens are critical steroid hormones that determine the expression of the male phenotype. Their actions are mediated by a single androgen receptor (AR) which, upon ligand binding, translocates to the nucleus to regulate the expression of androgen-responsive genes. Mutations that disrupt AR function totally result in the complete feminization of 46 XY individuals and the complete androgen insensitivity syndrome. Studies have revealed that AR mutations that do not lead to complete abrogation of its activity can cause a wide spectrum of milder androgen insensitivity syndromes, from ambiguous genitalia in newborn infants to 'idiopathic' male infertility. Recent studies indicate that missense amino-acid substitutions in the ligand-binding domain of the AR result in infertility through a novel mechanism that involves defective protein-protein interactions between receptor domains and coactivator proteins. Independent of missense mutations, studies involving Singaporean, Australian, North American and Japanese subjects indicate that increases in length of a trinucleotide repeat (CAG) tract, encoding a polyglutamine stretch in the transactivation domain of the AR, are associated with increased risk of defective spermatogenesis and undermasculinization. This association was however not observed in European populations, suggesting that the genetic background may play a significant role in the expression of the AR defects. PMID- 12638778 TI - The present and future state of hormonal treatment for male infertility. AB - Although male factors contribute to over half of all cases of infertility, most infertile men are described as 'idiopathic oligo/asthenozoospermic' rather than diagnosed precisely; hence, specific medical treatment is not possible. One uncommon but treatable cause of male infertility is gonadotrophin deficiency in which gonadotrophin replacement therapy is highly effective at inducing spermatogenesis and fertility. Hormonal therapy is a logical approach for empirical drug therapy given the fundamental role of hormonal regulation in spermatogenesis. However, treatment with GnRH analogues, gonadotrophins, androgens, anti-estrogens, aromatase inhibitors, growth hormone- and prolactin suppressing drugs is ineffective in unselected infertile men. Prolonged high-dose glucocorticoid therapy for sperm autoimmunity may improve pregnancy rates modestly, but the risks are generally unacceptable compared with IVF or ICSI. For these reasons, modern reproductive technologies, notably ICSI/IVF, have become the de-facto standard empirical treatment of male infertility, despite involving significant though infrequent risks to the fetus and mother. There remains a potential for hormonal methods to improve sperm quality or ultrastructure in subgroups of infertile men more responsive to hormonal manipulation or using novel protein or gene-targeted therapies or biochemical approaches based on post hormonal receptor mechanisms that stimulate spermatogenesis. How such novel hormonal methods will develop in conjunction with improved ICSI/IVF or cloning technologies, and the potential role of adjunctive hormonal therapy remains to be clarified. PMID- 12638779 TI - Stem cells: you can't tell a cell by its cover. AB - Embryonic stem (ES) cells are capable of unlimited self-renewal and have the ability to give rise to all tissue types in the body. The use of human ES cells for tissue and cell therapeutics has been suggested, but is limited by ethical concerns as these cells are derived from the inner cell mass of human embryos. In addition, the need for HLA matching of ES cell-derived tissues for allogeneic transplantation would require a bank of several thousand ES cell lines to make tissue therapeutics practical. Recently, adult stem cells-of which those in bone marrow are the best studied-have been shown to be capable of multilineage differentiation into cells of various non-blood tissues. Umbilical cord blood (UCB) haematopoietic stem cells have been shown to be equivalent to bone marrow stem cells for reconstitution of the haematopoietic system. Preliminary studies have also demonstrated that UCB haematopoietic stem cells are multipotent and capable of differentiating into non-blood cell types. This observation raises the exciting possibility of replacing human ES cells for tissue and cell therapeutics with UCB blood haematopoietic stem cells that are normally discarded with the placenta after delivery. PMID- 12638780 TI - Effects of in-vivo and in-vitro environments on the metabolism of the cumulus oocyte complex and its influence on oocyte developmental capacity. AB - There has been an improvement in the blastocyst rates achieved following in-vitro embryo production that can largely be attributed to improved embryo culture conditions based on an increased knowledge of the in-vivo environment, as well as the metabolic needs of the embryo. Despite this, in-vitro oocyte maturation (IVM) conditions have remained largely unchanged. Within the antral follicle, numerous events affect oocyte maturation and the acquisition of developmental competency, including: interactions between somatic cells of the follicle (in particular cumulus cells) and the oocyte; the composition of follicular fluid; and the temperature and vascularity of the follicular environment. Many of these factors change with follicle size and oocyte growth. In contrast, culture conditions for IVM are based on somatic cells that often do not reflect the follicular environment, and/or have complex compositions or additives such as macromolecule supplements that are undefined in nature. Metabolites included in media such as glucose, pyruvate, oxygen and amino acids have been shown to have differential influences on oocyte maturation and competency. Manipulation of these factors and application of gained knowledge of the in-vivo environment may result in improved in-vitro oocyte maturation and overall in-vitro embryo production. PMID- 12638781 TI - Implantable contraceptives for women. AB - Progestogen-only implantable contraceptives are used by increasing numbers of women worldwide. This review outlines the evidence accumulated on these methods to date. Reviews of toxicological evaluations, clinical trials, endocrinological, epidemiological and social science studies, as well as operations research and economic evaluation were undertaken in preparation for an Expert Consultation convened by the World Health Organization in 2001. At the meeting, these reviews were further evaluated and the research results summarized in this consensus paper. A large body of evidence demonstrates the high contraceptive effectiveness and safety of the 5-year levonorgestrel-releasing implants Norplant and Jadelle. Information on the 3-year etonogestrel-releasing implant Implanon is more limited, but suggests that this implant has a high contraceptive effectiveness and a satisfactory safety profile. Information available on levonorgestrel releasing implants manufactured and approved in China suggests that their clinical performance is satisfactory, but was insufficient to allow their full safety assessment. For all implants, there is insufficient information on their use by women with medical conditions. Provision of contraceptive implants requires good quality family planning services and specific provider training. PMID- 12638782 TI - Clinical management of low ovarian response to stimulation for IVF: a systematic review. AB - Poor response is not a rare occurrence in ovarian stimulation. Although not fully accepted, the most dominant criteria for poor ovarian response are small numbers of follicles developed or oocytes retrieved, and low estradiol (E2) levels after the use of a standard stimulation protocol. There is no ideal predictive test as the poor responder is revealed only during ovulation induction; however, increased levels of day 3 FSH and E2 as well as decreased levels of inhibin B can be used to assess ovarian reserve. Several protocols have been proposed for clinical management of low ovarian response in IVF. Although high doses of gonadotrophins have been used by the vast majority of authors, results have been controversial and prospective randomized studies have shown little or no benefit. The few available relevant studies do not indicate that recombinant FSH improves outcome. Flare-up GnRH agonist protocols (including all dosage varieties) produce better results than standard long luteal protocols. Luteal initiation GnRH agonist 'stop' protocols were shown to improve ovarian response according to prospective studies with historical controls, but this was not confirmed by well designed prospective, randomized, controlled studies. The few available data obtained with GnRH antagonists have not shown any benefits. Adjuvant therapy with growth hormone (GH) or GH-releasing factors results in no significant improvement. The use of corticosteroids reduces the incidence of poor ovarian response in women undergoing IVF treatment. The limited data obtained with nitric oxide donors are encouraging. Pretreatment with combined oral contraceptives prior to stimulation may help ovarian response. No benefit was observed with standard use of ICSI or assisted hatching of zona pellucida. Finally, natural cycle IVF has produced results which are comparable with those obtained with stimulated cycles in true poor responders. Well-designed, large-scale, randomized, controlled trials are needed to assess the efficacy of these different management strategies. PMID- 12638784 TI - Is exercise bad for the immune system? PMID- 12638783 TI - Review of clinical course and treatment of ovarian hyperstimulation syndrome (OHSS). AB - The ovarian hyperstimulation syndrome (OHSS) is a rare iatrogenic complication of ovarian stimulation occurring during the luteal phase or during early pregnancy. This complication is unusual as it is not the consequence of a treatment which is vital or mandatory for the patient's health. Nevertheless, it can be accompanied by severe morbidity and may even be fatal. Data pertaining to the clinical course and consequences of OHSS in women and its treatment were searched using Medline, Current Contents and PubMed. To date, only a few studies have collected a large number of cases of OHSS. The clinical course of OHSS may involve, according to its severity and the occurrence of pregnancy, electrolytic imbalance, neurohormonal and haemodynamic changes, pulmonary manifestations, liver dysfunction, hypoglobulinaemia, febrile morbidity, thromboembolic phenomena, neurological manifestations and adnexal torsion. Treatment of the acute phase relies only on an empirical and symptomatic approach. The general approach will be adapted to the levels of severity. Specific approaches such as paracentesis, pleural puncture, surgical approach of OHSS and specific medication during OHSS were evaluated sporadically. More adequate treatment methods would require a better understanding of the underlying pathophysiological mechanisms, to promote an aetiological therapeutic approach. Properly conducted studies, including large numbers of patients are required in order to determine the best method of prevention and management. PMID- 12638785 TI - Strangles, bastard strangles, vives and glanders: archaeological relics in a genomic age. PMID- 12638786 TI - Systemic concentrations of antioxidants and biomarkers of macromolecular oxidative damage in horses with grass sickness. AB - REASONS FOR PERFORMING STUDY: The aetiopathogenesis of equine grass sickness (EGS) is unknown. The role of free radical-mediated neuronal damage has not previously been investigated in this condition. OBJECTIVES: To investigate the potential contribution of oxidative damage and antioxidant status to neurodegeneration in EGS. METHODS: Systemic levels of surrogate biomarkers were determined in 10 horses with acute EGS and in 2 control populations; 10 healthy horses co-grazing with the 10 EGS horses at the onset of clinical disease, and 10 healthy mares grazing where EGS has not been reported. RESULTS: EGS horses had alterations in levels of several antioxidants, consistent with oxidative stress, the acute phase response and/or the secondary metabolic complications of EGS. EGS horses had elevated plasma dihydroxyphenylalanine (DOPA) levels. CONCLUSIONS: The elevated DOPA levels probably reflected a generalised disturbance of catecholamine metabolism rather than increased DOPA production via free radical mediated oxidation of tyrosine. However, there was no evidence of systemic macromolecular oxidative damage. POTENTIAL CLINICAL RELEVANCE: Further work is required to determine whether macromolecular oxidative damage occurring at the neuronal level contributes to EGS. PMID- 12638787 TI - Effects of imprint training procedure at birth on the reactions of foals at age six months. AB - REASONS FOR PERFORMING STUDY: While imprint training procedures have been promoted in popular magazines, they have received limited scientific investigation. OBJECTIVES: To determine the effects of a neonatal imprint training procedure on 6-month-old foals and to determine if any one session had a greater effect than others. METHODS: Foals (n = 131) were divided into the following treatments: no imprint training, imprint training at birth, 12, 24 and 48 h after birth or imprint training only at birth, 12, 24, 48, or 72 h after birth. Foals then received minimal human handling until they were tested at 6 months. RESULTS: During training, time to complete exposure to the stimulus was significant for only 2 of 6 stimuli. Percentage change in baseline heart rate was significant for only 2 of 10 stimuli. These 4 effects were randomly spread across treatments. CONCLUSIONS: Neither the number of imprint training sessions (0, 1, or 4) nor the timing of imprint training sessions (none, birth, 12, 24, 48, or 72 h after birth) influenced the foal's behaviour at 6 months of age. POTENTIAL CLINICAL RELEVANCE: In this study, imprint training did not result in better behaved, less reactive foals. PMID- 12638788 TI - Prolonged suppression of the innate immune system in the horse following an 80 km endurance race. AB - REASONS FOR PERFORMING STUDY: An increased susceptibility to bacterial and viral infections of the respiratory tract, which results in a loss of performance, has been reported in racehorses. Much research has focused on the influence of high intensity exercise of a short duration on immune system function in horses, but scant attention has been given to prolonged endurance exercise as an immune modulator. OBJECTIVES: The objective of this study was to evaluate the effect of an 80 km endurance race on the monocyte and neutrophil oxidative burst, serum cortisol, glutamine and plasma glucose concentrations in 8 endurance-trained horses (mean +/- s.d. age 9.4 +/- 2.2 years). METHODS: Blood samples were drawn from the horses prior to and following an 80 km ride. RESULTS: Mean time for completion of the 80 km race was 306 +/- 40 mins. Immediately post race mean serum cortisol concentration, blood monocyte and neutrophil counts were higher and blood lymphocyte counts and plasma glucose concentration were lower compared with prerace values (P < 0.05). Neutrophil and monocyte oxidative burst activity decreased following the race and had not regained prerace values after 3 days of rest (P < 0.05). CONCLUSIONS: The present study indicates that long duration exercise in horses has a negative impact on the function of the innate immune system that lasts several days post race. Precise mechanisms instigating the fall in innate immune system function are unclear and multifactorial, but may be attributed, at least in part, to a high serum cortisol response during very prolonged exercise. POTENTIAL CLINICAL RELEVANCE: A prolonged bout of exercise results in a long-term suppression of the innate immune system function in horses which may, in part, account for the observed increase of infectious episodes in horses during training. PMID- 12638789 TI - A retrospective case-control study of horse falls in the sport of horse trials and three-day eventing. AB - REASONS FOR PERFORMING STUDY: Serious injuries to horses and riders in horse trials (HT) and three-day events (3DE) are usually associated with falls of horses, which invariably involve falls of the riders. Many potential causes for these falls have been discussed. OBJECTIVES: The aim of this case-control study was to investigate the risk factors for horse falls on the cross-country phase of horse trials and three-day events. METHODS: Using retrospective data, significant risk factors identified with unvariable analysis (P value <0.2) were entered into a multivariable logistic regression model. Significant risk factors (P value <0.05) were included in the final model. RESULTS: It was revealed that a number of course, obstacle and rider variables were significantly and independently associated with the risk of falling. Falling was associated with obstacles sited downhill (Odds ratio [OR] 8.41) and with obstacles with ditches in front (OR = 5.77). CONCLUSIONS: The relationship between course variables and the risk of falling was characterised and showed a significantly increased risk with increasing numbers of jumps on the course and for jumping efforts later in the course. In contrast, after allowing for the total number of obstacles on the course, an increase in the total number of jumping efforts appeared to have a protective effect. A later cross-country start time was associated with a decreased risk of a horse fall. Amateur event riders were approximately 20 times more likely to fall than professional riders. POTENTIAL CLINICAL RELEVANCE: This study has identified a number of risk factors associated with horse falls and highlights areas that can be altered to improve safety in cross-country competitions. PMID- 12638791 TI - Equine melanoma in a population of 296 grey Lipizzaner horses. AB - REASONS FOR PERFORMING STUDY: Equine melanomas occur most commonly in grey horses at age 5 years or more. Generally, benign and malignant melanomas are distinguished by microscopy, but a more distinct classification would be helpful. OBJECTIVES: The objectives of this study were to gain further evidence concerning the occurrence of melanotic tumours, and to evaluate the impact of heredity on melanoma development. METHODS: A clinical study was conducted on a defined population of 296 grey horses of Lipizzaner breed. Individuals were classified according to their stage of disease using a 0-5 scale. Heritability was estimated on a sample of 296 grey horses with pedigrees traced back as far as 32 generations. RESULTS: Of the 296 horses, dermal melanomas were present in 148 horses (50%), 68 of which were more than age 15 years; 51 of these were melanoma bearing. In 75.6% of cases, melanotic tumours were detected underneath the tail. Although melanoma-bearing grey horses were encountered up to stage 4, none of the affected individuals suffered any severe clinical effect or was handicapped in performance. Statistical analysis revealed highly significant effects of stud and age (P < 0.0001), explaining 28% of the total variability. CONCLUSIONS: In contrast to melanomas in solid-coloured horses characterised by early metastases, melanomas in grey horses showed less malignancy. Affected individuals often had encapsulated nodules or structures similar to human blue nevi. Grey horse specific genetic factors inhibiting metastatic processes may be responsible for this phenomenon. POTENTIAL CLINICAL RELEVANCE: Although the obtained heritability estimate of 0.36 with a standard error of 0.11 indicates a strong genetic impact on the development of melanoma in ageing grey horses, a possible influence of the genes with large effects was also suggested. Therefore, further analysis is required of melanoma development in the ageing grey horse. PMID- 12638790 TI - Brain abscesses as a metastatic manifestation of strangles: symptomatology and the use of magnetic resonance imaging as a diagnostic aid. AB - REASONS FOR PERFORMING STUDY: The occurrence of unexpectedly high numbers of horses with neurological signs during two outbreaks of strangles required prompt in-depth researching of these cases, including the exploration of magnetic resonance imaging (MRI) as a possible diagnostic technique. OBJECTIVES: To describe the case series and assess the usefulness of MRI as an imaging modality for cases suspected of space-occupying lesions in the cerebral cavity. METHODS: Four cases suspected of suffering from cerebral damage due to Streptococcus equi subsp. equi infection were examined clinically, pathologically, bacteriologically, by clinical chemistry (3 cases) and MRI (2 cases). In one case, MRI findings were compared to images acquired using computer tomography (CT). RESULTS: In all cases, cerebral abscesses positive for Streptococcus equi subsp. equi were found, which explained the clinical signs. Although the lesions could be visualised with CT, MRI images were superior in representing the exact anatomic reality of the soft tissue lesions. CONCLUSIONS: The diagnosis of bastard strangles characterised by metastatic brain abscesses was confirmed. MRI appeared to be an excellent tool for the imaging of cerebral lesions in the horse. POTENTIAL CLINICAL RELEVANCE: The high incidence of neurological complications could not be explained but possibly indicated a change in virulence of certain strains of Streptococcus equi subsp. equi. MRI images were very detailed, permitting visualisation of much smaller lesions than demonstrated in this study and this could allow prompt clinical intervention in less advanced cases with a better prognosis. Further, MRI could assist in the surgical treatment of brain abscesses, as has been described earlier for CT. PMID- 12638792 TI - Risk factors associated with behavioural disorders of crib-biting, weaving and box-walking in Swiss horses. AB - REASONS FOR PERFORMING STUDY: Studies on the prevalence of behavioural disorders in horses and on associated risk factors have revealed inconsistent results. There are many studies on the neuropharmacological, surgical or mechanical therapy of stereotypies, but little is known about their causation. OBJECTIVES: To explore risk factors associated with the occurrence of behavioural disorders in horses. METHODS: A sample of horse owners, selected randomly and representative for Switzerland, was contacted in a postal survey. Answers were provided for 622 stables (response rate 35.2%). Individual data of 2,341 horses were examined with path analysis (multivariable linear and logistic regression), and adjustment made for possible confounding effects due to age and breed. RESULTS: Out of 60 possible risk factors, 11 were associated with the outcome at the univariable level (null-hypothesis path model) and 3 factors remained after the backward logistic regression procedure. Mature Warmbloods and Thoroughbreds, assessed by the owners to be reactive, fed 4 times a day and without daily pasture, had increased odds of displaying crib-biting, weaving and box-walking. Furthermore, indirect associations of 5 factors with the outcome were identified. CONCLUSIONS: The final logistic regression model of risk factors leads to the hypotheses that causal prevention of stereotypic behaviours should be based upon housing and management conditions which allow tactile contact with other horses (e.g. mutual grooming), daily free movement (paddock or pasture), as well as the provision of high amounts of roughage but of little or no concentrates. POTENTIAL CLINICAL RELEVANCE: It is one of the aims of population medicine to prevent the development of behavioural disorders. Further research is needed to test the concluding hypotheses in experimental studies or to verify them in the context of similar observational studies. PMID- 12638793 TI - Uptake of 5-hydroxytryptamine by equine digital vein endothelial cells: inhibition by amines found in the equine caecum. AB - REASONS FOR PERFORMING STUDY: 5-hydroxytryptamine (5-HT; serotonin) is a potent vasoconstrictor of equine digital blood vessels and has been implicated in the pathogenesis of acute laminitis. OBJECTIVES: The aims of this study were firstly to examine whether cells of the digital blood vessel wall exhibited an active uptake mechanism for 5-HT and to characterise its efficiency; and secondly, to study the potential inhibitory effect on this process of other amines, produced in the equine caecum. METHODS: Confluent monolayers of equine digital vein endothelial cells (EDVEC) and equine digital vein smooth muscle cells (EDVSMC) were incubated with [3H]5-HT (0.1-250 micromol/l) and the total and active uptake calculated. Equine pulmonary vein endothelial cells (EPVEC) were used as a positive control. RESULTS: Both EDVEC and EDVSMC showed uptake of [3H]5-HT by nonfaci litated diffusion; however, only EDVEC showed evidence of saturable facilitated uptake mechanism, with a Km of 41.6 +/- 9.3 micromol/l, which was significantly higher than that of EPVEC (9.9 +/- 2.1 micromol/l). All 6 caecally derived amines examined (tyramine, spermine, isoamylamine, tryptamine, phenylethylamine and isobutylamine) inhibited the total uptake of [3H]5-HT in a concentration-dependent manner, tyramine having the lowest IC50 (3.7 x 10(-6) mol/l). CONCLUSIONS: These data suggest that facilitated uptake into the endothelium could play a role in moderating the vasoconstrictor response to 5-HT in the equine digital circulation. POTENTIAL CLINICAL RELEVANCE: The vasoconstrictor action of 5-HT could be potentiated by gut-derived amines, providing a feasible link between GI disturbances and the pathophysiology of laminits. PMID- 12638794 TI - Effects of alpha-2 adrenoceptor agonists during recovery from isoflurane anaesthesia in horses. AB - REASONS FOR PERFORMING STUDY: Recovery from inhalant anaesthesia in the horse is a critical and difficult period to manage; however, several factors could help to obtain a calm recovery period including choice of anaesthetic and analgesic procedure used and the conditions under which anaesthetic maintenance and recovery occur. OBJECTIVES: The objective of this study was to evaluate and compare the quality of recovery in horses administered saline, xylazine, detomidine or romifidine during recovery from isoflurane anaesthesia. METHODS: Six mature and healthy horses were premedicated with i.v. xylazine and butorphanol, and anaesthesia induced using ketamine. After 2 h of inhalant anaesthesia with isoflurane vaporised in oxygen, saline solution, xylazine (0.1 mg/kg bwt), detomidine (2 microg/kg bwt) or romifidine (8 pg/kg bwt) were administered. The quality of recovery of each horse and the degree of sedation and ataxia were evaluated. Cardiovascular and respiratory parameters were recorded, and arterial blood samples obtained and analysed for pH, PO2 and PCO2 during recovery. RESULTS: Quality of recovery was better in groups treated with alpha-2 adrenergic receptors agonists, showing less ataxia. Degree of sedation was greater in the romifidine group. CONCLUSIONS: We concluded that the administration of alpha-2 adrenoceptor agonists during recovery from isoflurane anaesthesia in horses prolonged and improved the quality of recovery without producing significant cardiorespiratory effects. POTENTIAL CLINICAL RELEVANCE: Administration of alpha-2 adrenoceptor agonists after inhalent anaesthesia could prevent complications during the recovery period. PMID- 12638795 TI - Responses of horses in behavioural tests correlate with temperament assessed by riders. AB - REASONS FOR PERFORMING STUDY: Behavioural tests as well as observers' ratings have been used to study horses' temperament. However, the relationship between the ratings and the responses in behavioural tests has not yet been studied in detail. OBJECTIVES: The aim of the present study was to examine this relationship between ratings and responses. METHODS: Eighteen mature Swedish Warmblood horses were subjected to 2 behavioural tests, one relating to novelty (novel object test) and one to handling (handling test). Subsequently, 16 of these horses were ridden by 16 equally experienced students, having no former experience with the horses. Immediately after each ride, the students scored the horse for 10 temperamental traits using a line rating method. RESULTS: It was shown that for each temperamental trait all 16 riders agreed on the ranking of the horses (0.2127.0%). Forty eight (67.6%) of the 71 had gastritis, 17 (25.7%) had duodenitis, eight (11.3%) had oesophageal candidiasis, seven (9.9%) had bile reflux, six (8.5%) had reflux oesophagitis, six (8.5%) had ulcers (five duodenal, one gastric) and one (1.4%) had gastric cancer. Fourteen (19%) had endoscopically normal mucosa. The prevalence of H. pylori was 82.3% (32/38) in patients with antral gastritis. All ulcers and the cancer lesion (adenocarcinoma) were associated with H. pylori. Histological gastritis was found in 57 (81.8%) and was significantly associated with H. pylori. CONCLUSION: Although dyspepsia is common in diabetic outpatients at KNH, endoscopic findings and H. pylori status are not significantly different from those of non-diabetic population. PMID- 12638806 TI - In-vitro sensitivity of Plasmodium falciparum to chloroquine, halofantrine, mefloquine and quinine in Madagascar. AB - OBJECTIVE: To determine how sensitive Plasmodium falciparum is to the major antimalarial drugs in Madagascar. DESIGN: Assessment of Plasmodium falciparum isolates sensitivity to antimalarials, by use of the in-vitro radioisotope method. SETTING: Ankazobe and Saharevo in the foothill areas; and Toamasina and Tolagnaro in the coastal areas (between January 1998 and November 1999). SUBJECTS: Primary Plasmodium falciparum isolates from patients with uncomplicated malaria attack. RESULTS: Between January 1998 and November 1999, of the 293 in vitro tests done with at least one antimalarial, 70% (205/293) were interpretable. As there was no significant difference between results from the four study sites, the data have been expressed as a whole. All of the successfully tested isolates were sensitive to halofantrine (n = 56) and to quinine (n = 199), 5.8% (12/205) of the isolates were resistant to chloroquine and 2% (4/199) to mefloquine. The geometric mean IC50 was 0.3 microg/L for halofantrine (95% CI = 0.1-0.4 microg/L); 9.4 microg/L for chloroquine (95% CI = 7.3-10.8 microg/L); 3.8 microg/L for mefloquine (95% CI = 3.3-4.3 microg/L); and 26.8 microg/L for quinine (95% CI = 24.3-29.4 microg/L). The low positive correlation found between halofantrine and chloroquine IC50s (n = 56; r = 0.41, P = 0.002) suggests a risk of cross-resistance between these two drugs. CONCLUSION: The degree and frequency of chloroquine resistance in-vitro is stationary in Madagascar compared to previous results during the last decade. The in-vitro sensitivity of P. falciparum to quinine, mefloquine and halofantrine encourages the use of these drugs as alternative in case of chloroquine treatment failure. Nevertheless, it is important to maintain and to extend malaria and drug sensitivity surveillance in Madagascar. PMID- 12638807 TI - Laboratory methods for diagnosis and detection of drug resistant Mycobacterium tuberculosis complex with reference to developing countries: a review. AB - OBJECTIVE: To outline principles, advantages and limitations of the currently available laboratory methods for diagnosis and detection of drug resistance of Mycobacterium tuberculosis complex. DATA SOURCE: Published series of peer reviewed journals and manuals written on laboratory methods that are currently used for diagnosis and detection of drug resistance of Mycobacterium tuberculosis complex were reviewed using the index medicus, pubmed and medline search. Conventional bacteriological microscopy and culture, BACTEC, and molecular-based techniques were included. Basic principles, advantages and limitations of the cited techniques have been highlighted. CONCLUSION: Conventional bacteriological microscopy and culture are usually used for diagnosis of tuberculosis (TB) particularly in developing countries. However, their limited sensitivity, specificity and delayed results make this provision inadequate. Despite the development of quicker and more sensitive novel diagnostic techniques, their complexity and high cost has limited their use in many poor-resource countries. Due to the rapidly growing TB problem in these countries, there is urgent need to assess promising alternative methodologies in settings with high disease prevalence. PMID- 12638808 TI - Cellular and molecular interactions in HIV infections: a review. AB - OBJECTIVE: To review the cellular and molecular interactions between HIV and the host immune system that lead to full-blown AIDS. DATA SOURCES: Published reports on HIV/host interaction during a fifteen year period beginning from 1987. STUDY SELECTION: Only those studies involving humans and non-human primates were selected. The studies included original articles and state-of-the-art reviews covering in vivo and in vitro findings. DATA EXTRACTION AND SYNTHESIS: This article presents a critical review of the cellular and molecular mechanisms of HIV infection and their relationship to the onset of AIDS. CONCLUSION: HIV has elaborated diverse and somewhat complicated mechanisms for the subversion and evasion of the host immune defence strategies. These include escape through mutation, prolonged latency of the infection, masking of the viral envelope proteins, down-regulation of MHC-I and up-regulation of the Fas-ligand on infected cell surfaces. This review enhances our understanding of HIV/AIDS disease and presents a basis on which management strategies could be developed. PMID- 12638809 TI - Occupational risk of infection by human immunodeficiency and hepatitis B viruses among health workers in south-eastern Nigeria. AB - OBJECTIVE: To assess the occupational risk of infection by human immunodeficiency virus (HIV) as well as hepatitis B virus (HBV) among healthcare workers in south eastern Nigeria. DESIGN: Cross-sectional study. SETTING: Three tertiary health institutions in south-eastern Nigeria. SUBJECTS: Doctors, nurses, laboratory staff and cleaners. MAIN OUTCOME MEASURES: Observation of the availability and use of protective equipment and materials in the various departments of the hospitals. RESULTS: Materials and equipments needed for protective and hygienic practices (adequate water supply, protective clothing and availability of disinfectants) were inadequate in all hospitals. Where available, they were found to be inconsistently used. Health workers in the three institutions were thus constantly exposed unnecessarily to blood and other body fluids which might be potentially infectious as well as injury from used sharps. CONCLUSION: The risk of acquiring HIV and HBV infections by health workers in this region of Nigeria in the course of performing their duties is therefore still apparently high. Though distinct viruses, they share similar mode of transmission and risk factors. Use of personal protective equipment and adoption of standard hygienic practices among health workers must be encouraged. Supply of protective materials and equipment should be greatly improved. It is recommended that reduction of occupational risks among health workers using this approach should form part of control strategies for both infections in the country. PMID- 12638810 TI - Advance provision of oral contraceptives to family planning clients in Kenya. AB - OBJECTIVE: In sub-Saharan Africa, many family planning programmes do not encourage advance provision of oral contraceptives to clients who must wait until menses to initiate pill use. Since some resistance to advance provision of pills is due to provider fears that the practice may be harmful, we conducted a study in Kenya in 1997 to compare pill-taking outcomes between 20 "advance provision" clients and 280 "standard" clients. DESIGN: Prospective observational study. SETTING: Six family planning clinics in Central and Western Kenya. SUBJECTS: Women presenting as new clients at MOH family planning clinics. INTERVENTIONS: Researchers used prospective tracking to compare indicators of pill-taking success between non-menstruating clients given pills to carry home for later use and menstruating clients who began pill use immediately. MAIN OUTCOME MEASURES: Pill-taking outcomes such as side effects, compliance, knowledge, satisfaction, and a continuation proxy. RESULTS: Among clients returning for re-supply, those receiving advance provision of pills did no worse than, and often had superior outcomes to, their counterparts who started taking pills immediately after the clinic visit. CONCLUSIONS: Advance provision of pills, already practiced worldwide, is safe and feasible. Explicit mention should be made of advance provision of pills in national family planning guidance documents and training curricula in Kenya and throughout sub-Saharan Africa. PMID- 12638811 TI - Attitudes of medical students to induced abortion. AB - BACKGROUND: Unsafe abortion causes 13% of maternal deaths worldwide. Safe abortion can only be offered under conditions where legislation has been passed for legal termination of unwanted pregnancy. Where such legislation exists, accessibility of safe abortion depends on the attitudes of doctors and other healthcare workers to induced abortion. Medical students as future doctors may have attitudes to abortion that will affect the provision of safe abortion. Little is known about the attitudes of South African medical students to abortion. OBJECTIVES: To assess sexual practices and attitudes of medical students to induced abortion and to determine some of the factors that may influence these attitudes. DESIGN: A cross-sectional analytic study involving the self-administration of an anonymous questionnaire. SETTING: The questionnaire was administered to medical students at a small, but growing, medical school situated in rural South Africa. MAIN OUTCOME MEASURES: Demographic data, sexual practices and attitudes to induced abortion. RESULTS: Two hundred and forty seven out of 300 (82.3%) medical students responded. Their mean age was 21.81 +/- 3.36 (SD) years, and 78.8% were Christians, 17.1% Hindus and 2.6% Muslims. Although 95% of the respondents were single, 68.6% were already sexually experienced, and their mean age at coitarche was 17.24+/-3.14 (SD) years. Although overall 61.2% of the respondents felt abortion is murder either at conception or later, the majority (87.2%) would perform or refer a woman for abortion under certain circumstances. These circumstances, in descending order of frequency, include: threat to mother's life (74.1%), in case of rape (62.3%), the baby is severely malformed (59.5%), threat to mother's mental health (53.8%) and parental incompetence (21.0%). Only 12.5% of respondents would perform or refer for abortion on demand, 12.8% would neither perform nor refer for abortion under any circumstances. Religious affiliation and service attendance significantly influenced some of these attitudes and beliefs. CONCLUSION: Although many of the medical students personally felt abortion is murder, the majority are likely to perform or refer patients for abortion under certain circumstances; only about a tenth are likely to perform or refer patients for abortion on demand. PMID- 12638812 TI - Prevalence of major depression in deliberate self-harm individuals in Harare, Zimbabwe. AB - OBJECTIVE: To investigate the prevalence of major depression in deliberate self harm individuals. DESIGN: A cross sectional descriptive study. SETTING: Three tertiary health care centres in Harare, Zimbabwe. SUBJECTS: Three hundred and eighty seven deliberate self-harm consecutive subjects were recruited from July 1, 1997 to December 31, 1997 for this study. Diagnostic procedures included complete medical history, physical and neurological examinations. The DSM-IV criterion for major depression was used for the diagnostic purpose. MAIN OUTCOME MEASURES: The prevalence of major depression, number of attempts and recent life events, methods used, feeling in deliberate self-harm individuals. RESULTS: The prevalence of major depression was 20.7% (n=80, 95% CI=16.7-24.2) according to the DSM-IV criterion. Sixty-two percent (95% CI=57.2-66.8) of the subjects with DSH were aged < or = 30 years. Significant differences (P<0.05) were found between DSH subjects with and without major depression related to age, education, life events and number of previous attempts. CONCLUSION: The most predictive factors associated with DSH risks are age, depression, recent loss, divorced, loss of job and history of previous attempts. Further studies are needed to address some causes of DSH. PMID- 12638813 TI - Alternative methods for assessing growth in children under five years of age. AB - OBJECTIVE: To establish alternative methods for growth assessment in children under five years of age. DESIGN: A cross sectional study of healthy Malawian children. SETTING: Postnatal wards and under-five clinics at the Queen Elizabeth Central Hospital, Ndirande and Chilomoni Health Centres in Blantyre city, Malawi. MAIN OUTCOME MEASURES: Cephalic, thoracic and pelvic measurements and their indices were examined for assessment of growth in these children. Thoracic and pelvic measurements were found to be the most appropriate alternative parameters to weight. RESULTS: All measurements significantly correlated positively with weight but their indices did not. The degree of significance, however, was higher in pelvic and thoracic dimensions (P = 0.000) than in cephalic ones (P < 0.001 for maximal cephalic length (MCL) and P < 0.022 for maximal cephalic breadth (MCB), respectively. The MCL also showed a high correlation with age (R = 0.842, P = 0.002) but age only correlated moderately with the MCB (R = 0.569). The thoracic circumference (TC) and inter-nipple distance (IND) showed a very high positive correlation with age (R = 0.908, P = 0.005) and R = 0.870, P = 0.001), respectively. Similarly, a high positive correlation was shown with age by pelvic circumference (PC) and inter-spinous distance (ISD), (R = 0.891, P = 0.006 and R= 0.692, P = 0.027), respectively. Whereas the pelvic index positively correlated with age (R = 0.040), cephalic and thoracic indices significantly correlated negatively with age. Skull shapes were dolicocephalic in 43.6% of the children, mesocephalic in 31.1% and branchycephalic in 25.3%, a clear difference from Caucasian children reported earlier by other authors. CONCLUSION: We have established that cephalic, thoracic and pelvic dimensions significantly correlate positively with weight as age does but their indices remained relatively constant. More importantly, thoracic and pelvic measurements were the most appropriate alternative methods for growth assessment in our studied population. PMID- 12638815 TI - Cigarette smoking and khat chewing among university instructors in Ethiopia. AB - OBJECTIVE: To assess the prevalence and risk factors of cigarette smoking and khat chewing. DESIGN: College based cross sectional. SETTING: Four colleges found in north west Ethiopia namely Gondar College of Medical Sciences, Gondar College of Teachers Education, Bahr Dar University Engineering Faculty, and Bahr Dar University Education Faculty. SUBJECTS: All instructors in these colleges. MAIN OUTCOME MEASURES: Prevalence of cigarette smoking and khat chewing. RESULTS: Seventy six (42.0%) instructors were either lifetime smokers or lifetime khat chewers or both. The current prevalence rates of cigarette smoking and khat chewing were found to be 13.3% and 21.0%, respectively. The majority of the instructors started smoking (56.8%) and khat chewing (40.0%) while they were senior high school or first year college students. Most of the instructors (82.1%) knew that cigarette smoking is a risk factor for lung diseases including lung cancer. Statistically significant difference (p<0.05) was seen between the outcome variable and the independent variables faculty, religion, education status, income and family history of khat chewing. CONCLUSION: The prevalence of cigarette smoking seemed to decrease among university instructors but the prevalence of khat chewing is almost the same as it was some years ago. Instructors knew the common health risks associated with cigarette smoking. The high schools and colleges should inform their students about the health and socioeconomic problems associated with cigarette smoking and khat chewing. Additionally, college students need counseling service on ways of coping with their problems. PMID- 12638814 TI - Bacteriological qualities of indoor and out-door drinking water in Kibera sub location of Nairobi, Kenya. AB - OBJECTIVES: To compare the bacteriological quality of out-house (tank or standpipe) water and in-house drinking water (storage containers) and determine the risk factors influencing it. DESIGN: A cross-sectional study. SETTING: The study was carried out in Kibera slums located 7 km southwest from the Nairobi City centre. SUBJECTS: Water samples from twenty outside tanks/standpipes and sixty from in-house water storage containers. MAIN OUTCOME MEASURES: Pour plate method was used to enumerate total bacterial counts in water, while the multiple tube technique was used to determine faecal coliform (FC) and faecal streptococci (FS) numbers. A questionnaire and environmental observation were used to determine the risk factors influencing bacteriological quality of water. RESULTS: The mean total bacterial counts (TBC) for out-house water was 46.6 per 100 ml while that for in-house water was 818.2 per 100 ml. Faecal coliforms were isolated from 7 (35%) standpipes and 57 (95%) in-house storage containers. The mean faecal coliform count was 93 and 103.4 per 100 ml for out-house and in-house water, respectively. The counts were significantly higher in the latter. Faecal streptococci were isolated from 2 (10%) standpipes and 37 (61.7%) in-house storage containers. The mean faecal streptococci counts were 35 and 65 per 100 ml for out-house and in-house water sources, respectively. Escherichia coli was isolated in 2 (10%) of out-house water and 30 (50%) of in-house. Of these, four were enteropathogenic, serotype 011 from one out-house water source and serotypes 011, 011, 0112ac from in-house water sources. CONCLUSIONS: Bacteriological contamination of water at the source with a further deterioration between the collection points and homes was observed. A defective water delivery system and inadequate environmental sanitation were a potential source of contamination for out-house water. Scoops were a major source of contamination for stored water. PMID- 12638816 TI - Herpes zoster myelitis: report of two cases. AB - Two male patients aged 40 and 45 years with HIV infection and paraplegia are presented. The two had sub-acute onset paraplegia with a sensory level, which developed 10 days after herpes zoster dermatomal rash. They both had asymmetrically involvement of the lower limbs. Investigation including imaging of the spinal cord did not reveal any other cause of the neurological deficit. The two responded very well to treatment with acyclovir. Herpes zoster myelitis is a condition likely to rise with the upsurge of HIV infection and there is a need to identify the condition early. We also review the literature on the subject. PMID- 12638817 TI - Ageing population in Africa and other developing communities: a public health challenge calling for urgent solutions. PMID- 12638818 TI - Informal carers of mentally infirm elderly in Lancashire. AB - OBJECTIVES: To investigate the determinants of satisfaction in caregiving and to compare satisfaction in care-giving amongst carers of demented and non-demented mentally infirm elders; and, assess carer attitudes and concerns, and their implications on care in the community. DESIGN: Cross-sectional study of informal carers of the elderly referred to a psychogeriatric service, using a questionnaire investigating carer satisfaction (CASI), care-recipient dependency needs, carer burden (CADI), carer concerns and attitudes in relation to caregiving, and the 28-item GHQ. SETTING: Lancashire communities of Fleetwood, Thornton-Cleveleys, Poulton-Le-Fylde, and Over-Wyre. RESULTS: Carers achieved significant degrees of satisfaction in their role as care-givers; there was no significant difference in the degree of satisfaction gained by carers of the demented and non-demented. The mean CASI score, for carers of the demented and non-demented was 23(5.5) and 24.4 (5.7) respectively (mean difference -2.9; CI 4.6, 0.1; p = 0.058). Dissatisfaction in care giving was determined by total burden (CADI) scores, and younger carer age. Emotional distress in carers was weakly inversely correlated with CASI scores (r = -0.21, p = 0.042). Concerns expressed by carers, included desire for information on care recipient disability (39.5%) and fear of nursing/residential home placements (43%). Most carers had a generally positive attitude to care giving, inspite of significant degrees of burden to which they were subjected. CONCLUSIONS: Carer-related factors, particularly younger age, rather than dependency factors, were determinant of care giving satisfaction. Greater involvement of older persons in care giving should be encouraged, with younger persons assisting if care giving becomes overbearing. Carers require education on care-recipient disabilities and the benefits of care in formal care institutions. PMID- 12638819 TI - Burden of informal carers of mentally infirm eldery in Lancashire. AB - OBJECTIVES: To compare the burden of supporting demented and non-demented mentally infirm elders in an English community; determine the prevalence of emotional distress in carers and to investigate the relationship between carer well being and duration of care giving. DESIGN: Cross-sectional study of informal carers of the elderly referred to a psychogeriatric service, using a questionnaire investigating carer socio-demography, carer-dependant interpersonal relationship in relation to care giving and care-recipient dependency needs using a modified version of the CADI; and, the 28-item GHQ. SETTING: Lancashire communities of Fleetwood, Thornton-Cleveleys, Poulton- Le-Fylde and Over-Wyre. RESULTS: Ninety one (72%) carers of the elderly mentally infirm participated in the study. Carers (n = 48) of the demented experienced significantly more burden, including psychological and physical health problems than carers (n = 43) of the non-demented (P = 0.001). The prevalence of emotional distress in all carers was 42% (dementia supporters = 56%, non-dementia supporters = 26%). Emotional distress in supporters was directly related to the degree of difficulties (particularly lack of private time, loss of control in caregiving tasks, patient behavioural problems) experienced in care giving, and, the degree of patient dependency needs. Carer/patient interpersonal relationship tended to worsen as care giving progressed; however, no significant association was established between duration of care and emotional distress in carers. Out of 51 carers receiving respite admission services, six (12%) considered such admissions as additional burden. CONCLUSIONS: Carer burden, including psychological and physical health concerns, was comparatively greater in carers of the demented than in carers of the non-demented. Respite care services although beneficial to most care givers, may constitute further burden to some. Factors accounting for these observations are explained. To ameliorate carer burden, measures enabling greateravailability of private time, and, improving care giving skills, should be encouraged. PMID- 12638820 TI - Substance use amongst secondary school students in rural and urban communities in south western Nigeria. AB - OBJECTIVES: To determine the prevalence and pattern of drug use among secondary school students in rural and urban communities in south western Nigeria. DESIGN: A survey of senior secondary school students for drug use using a modified form of the World Health Organisation (WHO) questionnaire for student drug use surveys after a pilot study. SETTING: Rural and urban communities in two local government areas in Ilesa, Osun State in south western part of Nigeria. SUBJECTS: A total of 600 students drawn from six secondary schools. MAIN OUTCOME MEASURES: Prevalence rates of drugs use, drug use pattern by sex, class, urban/rural location of school and age of initiation into drug use. INTERVENTION: Determination of drug use prevalence rates and pattern under anonymous condition. RESULTS: Questionnaires of 567 respondents were analysed. Their mean age was 17.0 years (S.D +/- 1.69). The most commonly used drugs and their current prevalence rates were salicylate analgesics, 48.7%; stimulants, 20.9%; antibiotics, 16.6%; alcohol, 13.4%; hypnosedatives, 8.9% and tobacco, 3.0%. Current and lifetime use of alcohol as well as current, past and lifetime use of tobacco occurred significantly more commonly among the males. Past and lifetime use of tobacco was significantly more common in the rural school. For the majority of respondents, initiation into drug use started at a very early age (14 years or below). The majority were mild current users of the drugs, except tobacco for which the majority were daily users. CONCLUSION: Secondary school students in south western Nigeria engage in use of commonly available drugs like their counterparts elsewhere. Stringent measures should be taken to effect supply and demand reduction. PMID- 12638821 TI - Socio-demographic characteristics of adolescent post-abortion patients in Blantyre, Malawi. AB - BACKGROUND: Unsafe sex, unwanted pregnancy and unsafe abortion are some of the emerging adolescent reproductive health challenges in Malawi. OBJECTIVES: To study socio-demographic characteristics of adolescent post-abortion patients and that of their male partners, with the aim of better understanding the determinants of adolescent fertility in Malawi, thus facilitating design of appropriate intervention strategies. DESIGN: A descriptive, cross-sectional study. SETTING: The Queen Elizabeth Central Hospital, Blantyre, Malawi. SUBJECTS: A total of 446 out of 465 eligible adolescents managed for incomplete abortion during the study period were enrolled from January 1st to December 31st 1997. RESULTS. Their mean age was 17.5 years (SD +/- 1.3), while that of their male partners was 22.4 years (SD +/- 3.4). The unmarried adolescents comprised 43.9% and students 38.6%. About 30.0% had attained secondary school level of education. The mean age at sexual debut was 15.7 years (SD +/- 1.75), about a year after that at menarche (14.3 years, (SD +/- 1.4)). The mean number of sexual partners they had had was 1.5 (SD +/- 0.86), the highest being 10. The index pregnancy was reportedly unwanted in 45. 1% of the total. The rate was higher among the young, single and students. They were also less likely to have informed their partners about the pregnancy or the abortion (p = 0.0001). About 52.8% of the female students were involved with fellow male students. Of the married adolescents, 4.4% reported having had extramarital affairs. CONCLUSION: There is need to design appropriate programmes to promote safe sex and reduce unwanted pregnancy and unsafe abortion with its potential sequelae among adolescents in Malawi. PMID- 12638822 TI - Trans-hiatal oesophagectomy as palliative treatment for carcinoma of the oesophagus. AB - OBJECTIVE: To determine the role of palliation with trans-hiatal oesophagectomy in Nigerian patients with carcinoma of the oesophagus. DESIGN: Prospective case series. The first series was from February 1986 to September 1987 (Series A) while the second series was from March 1989 to November 1996 (Series B). SETTING: Cardiothoracic Surgery Unit (CTSU) of the University College Hospital, Ibadan, Nigeria. SUBJECTS: First series consisted of 10 consecutive operable patients with carcinoma of oesophagus seen over the period of study. The second series consisted of 21 consecutive patients with same disease. INTERVENTION: All patients had transhiatal oeosphagectomy by a two team approach and immediate placement of the freed stomach in the posterior mediastinum and cervical oesophagogastrostomy. RESULTS: Patients in both series had a comparable age range of 43 - 80 years for series A and 40 - 82 years for Series B. The duration of symptoms were 2 - 6 months and 2 - 12 months respectively, for series A and B. In series A, nine patients had carcinoma of the middle-third (M1/3) of the thoracic oesophagus and one patient had carcinoma of lower-third (L1/3) of the thoracic oesophagus. In series B, 18 patients had M1/3 and three patients had L1/3 lesions. Average blood loss in series A was 1,067 mls, corresponding value for series B was 852 mls. Postoperatively, all cases were classified as stage III or stage IV disease. There were 18 complications in eight patients in series A and 22 complications in 10 patients in series B. The commonest complications in series A were pleural enteries in six patients, haemorrhage four patients (three intraoperative, one post-operative) and respiratory failure (two patients). The commonest in series B were pleural enteries in nine patients, anastomotic leaks and stenosis in four patients and respiratory failure in three patients. Hospital mortality was 50% in Series A and 14.3% in series B. The causes of death were haemorrhage and respiratory failure in series A, respiratory failure in series B. Survival period in series A of the five patients discharged was for a median of 84 months, for series B, four patients were alive at 18 months post-operative, one patient attended follow-up clinic 24 months after surgery. No other adjunctive therapy was offered to the patients. CONCLUSION: Trans-hiatal oesophagectomy is a procedure suitable for patients with carcinoma of the oesophagus and affords palliation at an "acceptable price" among carefully selected patients with advanced carcinoma of the oesophagus. PMID- 12638823 TI - Anatomy and pathology of coronary artery in adult black Kenyans. AB - OBJECTIVE: To determine the pattern of coronary arterial anatomy and prevalence of postmortem coronary pathology in adult Kenyan Africans. DESIGN: Cross sectional study. SETTING: The Nairobi City Mortuary and the Department of Human Anatomy, University of Nairobi. METHOD: One hundred hearts were retrieved during consecutive autopsies over a three month duration and systematically dissected. Details on coronary ramification, dominance, atherosclerosis, tunnelling and hypoplastic segments were obtained and statistically analysed. RESULTS: Seven patterns of left coronary ramifications were identified. The right coronary artery anomalously exited from the left coronary sinus in one situation. There were separate ostia for the coronary artery branches in 2% and 31 % of cases on the left and right coronary systems respectively. The right coronary artery was dominant in 82% of the hearts. Coronary ostial sizes and luminal dimensions showed wide variations. Only two of the hearts had atheromatous luminal narrowing greater than 75% of the cross-sectional area. Muscle bridges of average depths of 1.1-2 mm were demonstrable in 29% of the autopsies. Diminutive left anterior descending artery was present in four cases. The right coronary artery was diminutive in one case. CONCLUSION: Coronary atherosclerosis is still a rarity in the setting within which the study was undertaken. The diverse patterns of ramifications of the coronary tree begs for caution during coronary investigations and interventional procedures. Coronary arterial anomalies, myocardial bridges, atheroma and diminutive arteries should be considered in cases of sudden cardiac death in the absence of other pathologies. PMID- 12638824 TI - Risk awareness, exposure to oral health information, oral health related beliefs and behaviours among students attending higher learning institutions in Dar es Salaam, Tanzania. AB - OBJECTIVE: To assess the level of exposure to information about oral health from local sources, risk awareness, oral health beliefs and behaviours among male and female students attending higher learning institutions in Dar es Salaam. DESIGN: A cross sectional survey. SETTING: Government recognised higher education institutions in Dar es Salaam, Tanzania in 1999. SUBJECTS: The study population comprised of 1123 university students in Dar es Salaam. MAIN OUTCOME MEASURES: Level of risk awareness, exposure to oral health information, oral health beliefs and behaviour among students. RESULTS: Despite apparent misconceptions, the level of risk awareness was generally high and did not differ between male and female students. Univariate analysis of variance revealed that female students were exposed to more oral health information, had stronger oral beliefs and performed preventive behaviours more frequently than did their male counterparts. In multiple linear regression analysis, beliefs about the importance of preventive behaviours were significantly related to the frequency with which such habits occurred (beta = 0.19, p < 0.001) across gender and students status. This relationship was significantly stronger among females (beta = 0.33, p < 0.001) than among males (beta = 0.11 p < 0.05). CONCLUSION: The results indicate that strengthening beliefs in the presence of accurate knowledge might successfully promote students' oral health enhancing behaviour. PMID- 12638825 TI - Management of oesophageal foreign bodies in children. AB - OBJECTIVE: To evaluate the pattern and outcome of management of retained oesophageal foreign bodies in children. DESIGN: Retrospective study. SETTING: Obafemi Awolowo University Teaching Hospitals Complex, Ile-Ife, January 1991 to December 2000. SUBJECTS: One hundred and eight paediatric patients, aged 0-14 years managed for retained oesophageal foreign bodies. INTERVENTION: The foreign bodies were removed endoscopically, using either a larynoscope or a rigid oesophagoscope, under general anaesthesia. RESULTS: There were 108 patients, with a mean age of 3.0 +/- 0.8 years and age range of 23 days to 14 years. The male to female ratio was 1.5: 1. Sixty four (59.3%) patients were within 1-5 year age group. Coins constituted 79.6% of the retained foreign bodies. Over 90% of patients presented within the first four days of the incidence. The dominant complaints were drooling of saliva in 37 (40.2%) and difficulty in swallowing in 35 (38.1%) patients. The commonest sites of the foreign body retention were the upper third of oesophagus 52(48.2%) and the hypopharynx 36(33.3%). The main complications following the foreign body removal were oesophageal perforations 4(3.7%) and lacerations 16(15%). CONCLUSION: Retained oesophageal foreign body is a common childhood health hazard. Early diagnosis and prompt skillful removal are necessary for a satisfactory outcome. PMID- 12638826 TI - Herbal medicine in Kenya: evidence of safety and efficacy. PMID- 12638827 TI - Prophylactic effect of multi-herbal extract 'Agbo-Iba' on malaria induced in mice. AB - OBJECTIVE: To determine the efficacy of a multi-herbal preparation extract of 'Agbo-Iba' on rodent malaria induced in mice. DESIGN: An experimental design in which mice were divided into four groups A,B,C,D representing control, prophylactic, chloroquine and 'Agbo-Iba' groups respectively. Each mouse was intraperitoneally inoculated with Plasmodium yoelii nigeriensis and treated with oral herbal extract or chloroquine syrup depending on group. SETTING: College of Medicine of the University of Lagos Medical Microbiology and Parasitology Laboratory. SUBJECTS: One hundred and twenty male and female albino mice aged 10 12 weeks with an average weight of 25 grams. MAIN OUTCOME MEASURES: The herbal extract was effective, preventing the development of parasitaemia in the prophylactic group of mice. RESULTS: After intraperitoneal inoculation of Plasmodium yoelii nigeriensis, a prepatent period of two days was observed before parasitaemia was established in all but the prophylactic group of mice. Induced infection was promptly aborted with oral chloroquine treatment in group C, while in groups A and D, infection terminated fatally. Group B mice appeared normal throughout the duration of investigation with 100% survival rate. CONCLUSION: 'Agbo-Iba' extract has some prophylactic action against malaria induced in mice with no apparent significant side effects. PMID- 12638828 TI - Missed opportunities and caretaker constraints to childhood vaccination in a rural area in Uganda. AB - BACKGROUND: Despite concerted support to vaccination programmes, coverage remains low. While health service reasons for this are known, there is little information on caretaker constraints to vaccination in Africa. OBJECTIVE: To establish the prevalence of missed vaccination opportunities and caretaker constraints to childhood vaccinations. DESIGN: Cross-sectional descriptive study. SUBJECTS: Caretakers of 408 children aged 12-23 months were interviewed. SETTING: Kiyeyi, a rural area in Eastern Uganda. RESULTS: Complete vaccination coverage by card was 26.7% while by history and card it was 44.6%. Of the 215 eligible children who sought treatment in a health facility where vaccination could be offered, 59.6% missed an opportunity to be vaccinated while 24.4% of the children missed an opportunity during routine vaccination sessions. Reasons for non-completion of vaccination included caretaker 'not bothered', being busy, or ill and fear of rude health workers. While most caretakers were aware of vaccination and its benefits, none knew the immunisation schedule. The major caretaker constraints were low level of formal education, fear of vaccine side effects, and perceived contraindications to vaccinations. CONCLUSION: Promotion of formal education for girls and educating mothers and health workers on the timing of vaccinations, their side effects and management might contribute to higher vaccination coverage. PMID- 12638829 TI - Prevalence of vitamin A deficiency among pre-school and school-aged children in Arssi Zone, Ethiopia. AB - OBJECTIVE: To investigate vitamin A status of pre-school and school aged children in the study area. DESIGN: Cross-sectional. SETTING: Arssi, Ethiopia. SUBJECTS: Four hundred and two children. RESULTS: Night blindness, Bitots spot, corneal xerosis, corneal ulceration and corneal scar were observed in 7.2%, 2.2 %, 0.2%, 0.5%, and 0.5% of the children respectively. The prevalence of xerophthalmia was higher in school aged children than pre-school children (P< 0.0001). Based on the WHO recommended cut-off level, serum retinol levels were in the "low" range (<20 microl/dl) in 51% of the children. CONCLUSION: The results indicate that vitamin A deficiency (VAD) is a public health problem in Arssi, with higher prevalence among school aged children than pre-school children. PMID- 12638830 TI - Irritable bowel syndrome in Kenyans. AB - OBJECTIVE: To study the prevalence and clinical presentation of irritable bowel syndrome (IBS) in black Kenyans. DESIGN: A retrospective study. SETTING: Nairobi Hospital, an urban private institution. SUBJECTS: Case files of all patients presenting with abdominal pain. MAIN OUTCOME MEASURES: Prevalence of IBS using modified Manning's Symptomatic criteria in 223 consecutive adult patients presenting with abdominal pain and discomfort who had normal clinical findings and normal general and gastrointestinal investigations. RESULTS: Out of the 3472 patient files screened, there were 281 patients who fitted the diagnosis of irritable bowel syndrome giving an overall prevalence of 8%. The mean age was 40 years with range of 11 to 75 years with a peak prevalence in the 3rd decade. The male to female ratio of 1.4:1 being statistical significant at a p-value of 0.010. There was no statistical significant difference between the males and females in constipation and diarrhoea at p-value of 0.84 and 0.82 respectively. Fifteen percent of the patients were either on antidepressants or had been seen by a psychiatrist. Nine percent had undergone laparatomy for the abdominal pain. CONCLUSION: The Black African patient is equally as exposed to irritable bowel syndrome as is his counterpart in the western world and has similar morbidity patterns. The syndrome may account for over 10% of patients with abdominal pain presenting to gastroenterologists in Africa. Proper attention to symptomatology may lead to clinching the diagnosis without resorting to many unnecessary and expensive investigations. PMID- 12638831 TI - Anthrax revisited. AB - BACKGROUND: Anthrax is an ancient disease affecting animals and humans. Sporadic cases of anthrax and small epidemics have been seen from time to time in different parts of the world and in Africa. However many clinicians are not very familiar with the various presentations and management of anthrax. It is relevant for the health care workers to re-familiarise themselves with all aspects of anthrax, with the impending threat of bioterrorism. OBJECTIVE: To familiarise healthcare workers on all aspects of anthrax. STUDY SELECTION: To describe epidemiology pathogenesis, clinical features, management and prevention of anthrax including measures to take when weapons grade anthrax is suspected. DATA SYNTHESIS: Three forms of the disease are recognised, cutaneous, inhalational and intestinal. Cutaneous anthrax is the most common form. Inhalation anthrax is the most severe form of anthrax. The treatment of anthrax in most cases is penicillin, however with the threat of bioterrorism, intentional releases of anthrax spores in the environment has caused much concern. Weapons grade anthrax of more virulent strain and resistant to commonly used antibiotics is possible. CONCLUSION: In view of the different clinical presentations and outcomes it is important that health care workers re-familiarise themselves with the disease and in the event of bioterrorism are able to take appropriate measures. PMID- 12638832 TI - Review of eight cases of insulinoma. AB - OBJECTIVE: To review patients records operated with the diagnosis of insulinoma and to discuss their clinical presentations, diagnostic and therapeutic modalities. DESIGN: Retrospective study. SETTING: Ankara Numune Teaching and Research Hospital, Turkey. SUBJECTS: Eight cases were operated in the Department of 6th Surgery, Ankara Numune Teaching and Research Hospital between 1994 and 2000. All patients had neuroglycopenic symptoms. Six patients had blood glucose levels of lower than 50 mg/dL during the admission. The other two patients had hypoglycaemia in the prolonged fasting test. Serum insulin/glucose ratio was diagnostic in all patients except one. Abdominal ultrasonography and computerised tomography could successfully localise the tumour in one case. In six patients tumours could be localised by endoscopic pancreatic ultrasonography. In one patient none of the studies could localise the tumour. Three tumours were located at the pancreatic head, one in the neck, two at the body and two at the tail. All tumours except one were palpable. Enucleation was the procedure of choice in four cases and distal pancreatectomy was the procedure of choice in four. RESULTS: Post-operative course was uneventful in seven patients. One patient died due to intra-abdominal sepsis. Hypoglycaemia was controlled in all patients after the surgery. CONCLUSION: Surgery is the mainstay of treatment of insulinoma. Enucleation should be the procedure of choice if possible. Endoscopic pancreatic ultrasonography has promising results and may replace invasive angiographic studies in the future. PMID- 12638833 TI - Surgical admissions to the Rift Valley Provincial General Hospital, Kenya. AB - OBJECTIVE: To describe the pattern of surgical admissions to the Rift Valley Provincial General Hospital and in particular the epidemiologic characteristics of trauma admissions. DESIGN: Retrospective study. SETTING: Rift Valley Provincial General Hospital, Nakuru from (1st January 1998 - 31st December 1999). SUBJECTS: All patients admitted to the various surgical wards (excluding eye patients) during the stated period and whose medical records were available and complete. METHODS: Medical records of all surgical patients admitted during the period of study were retrieved using admission data from casualty, surgical out patient and Annex Hospital registers. Further medical data were obtained from wards admission registers, nurses report books and records from theatre and radiology books. A special data-form was used to collect the required information. The data was subjected to simple statistical analysis. RESULTS: There were 5,907 surgical admissions of whom 3,411 cases seventy three point five percent were trauma admissions, 1,499 cases (25.4%) were non-traumatic emergency surgical admissions and 997 cases (16.8%) were elective surgical admissions. 73.5% of all trauma admissions were males and 57.6% were in the 21-60 year age group. The most common injuries were soft-tissue injuries, fractures, burn injuries and head injuries, while the leading causes of trauma were road traffic accidents (32.7%), assaults (23.8%), falls (15.5%) and burns (13%). Fifty one point seven percent of all performed surgical operations were trauma-related. The mean length of hospital stay for trauma patients was 10.4 days. Trauma was the leading cause of death (6.6%) among all surgical admissions. CONCLUSION: Trauma, particularly due to road traffic accidents and violence is a growing public health problem in this region that urgently calls for specific intervention measures. Further studies of disability levels as well as costs of trauma care are recommended. PMID- 12638834 TI - Evaluation of 25-gauge Quincke and 24-gauge Gertie Marx needles for spinal anaesthesia for caesarean section. AB - OBJECTIVE: To compare the insertion characteristics and rate of complications between 25-gauge Quincke and 24-gauge Gertie Marx needles. DESIGN: Prospective, randomized study. SETTING: University of Benin Teaching Hospital; a university affiliated tertiary centre. SUBJECTS: Parturients (ASA 1 and 2) scheduled for elective caesarean section. They were randomly assigned to receive spinal anaesthesia with either 25-gauge Quincke needle or 24-gauge Gertie Marx needle. The patients with abnormal spaces, coagulopathy, infection, pre eclampsia/eclampsia or obesity were excluded. MAIN OUTCOME MEASURES: The number of attempts at successful identification of the spinal space, intraoperative complications, incidence of postdural puncture headache (PDPH), non-postdural puncture headache (NPDPH) and backache. RESULTS: Sixty women were studied. The 24 gauge Gertie Marx needle resulted in more successful location of the spinal space on the second attempt (P<0.05). Non-postdural puncture headache was seen in 43% of the study population. PDPH was seen in 10% of the Quincke group and none in the Gertie Marx group. There was no difference in the incidence of backache in both groups. CONCLUSION: The ease of insertion and low incidence of PDPH with the Gertie Marx needle may encourage trainee anaesthetists to use this needle for caesarean section. PMID- 12638835 TI - Childcare practices of commercial sex workers. AB - OBJECTIVE: To determine the childcare practices of commercial sex workers (CSWs). DESIGN: A descriptive cross-sectional survey was conducted between July and December 2000 during which a structured questionnaire was administered. SETTING: Kibera slum, Nairobi, Kenya. SUBJECTS: Three hundred eighty five CSWs and four focus group discussions (FGDs) held. Health cards from 126 under five years old children belonging to the respondents were reviewed for immunization status and regularity of growth monitoring. RESULTS: The mean age of the 385 CSWs surveyed was 32 +/- 7 years and mean duration of sex work was 6 +/- 4 years. The mean number of living children was 3.4 +/- 2 and 81.2% of the mothers lived with their children. Three quarters of the CSWs practised prostitution at home. The most common daily childcare activities by the mothers were food preparation (96.2%) and washing children's clothes (91.3%). Overall 96.8% of their under-five years old children were fully immunized and 80% of their under one year old children had their growth monitored monthly. About three quarters of the mothers with adolescent children educated them on HIV/STDs. Health seeking behaviour for the children was hampered by health care cost (71.4%) and consumption of alcohol by the mothers. Like other mothers, the CSWs encouraged their adolescent children to take up some adult roles such as maintaining a clean house (93.3%). However only 2.0% took time to converse or counsel the children. Focus group discussions (FGDs) with the CSWs showed that children were left unattended at night while the mothers went out in search of clients. Efforts to provide better education for the children were undermined by lack of funds (52.2%) and truancy (46.6%). One third of the study population had invested for the future maintenance of their children. CONCLUSION: There was more emphasis on physical, rather than psychological aspect of childcare. The practice of living with the children ensured that earnings from the sex trade were used for the immediate needs of the children such as food. However this practice had a negative influence on the children as the majority of the respondents conducted their sexual business at home with little or no privacy. Health seeking behaviour for the children was hampered by lack of funds and to some extent alcohol consumption by the mothers. Efforts to invest in the education of their children were undermined by lack of funds and truancy. PMID- 12638836 TI - Rhinocerebral mucormycosis: case report. AB - Rhinocerebral mucormycosis is a rare, fulminating opportunistic fungal infection caused by a fungus of the order mucorales. These fungi are ubiquitous, subsisting on decaying vegetation and diverse organic material. Although the fungi and spores of mucorales show minimal intrinsic pathogenicity towards normal persons, they can initiate aggressive and fulminating infection in the immune compromised host. Because rhinocerebral mucormycosis occurs infrequently it may pose a diagnostic and therapeutic dilemma for those who are not familiar with its clinical presentation. We present a patient with classical presentation of rhinocerebral mucormycosis involving the paranasal sinuses, the orbit and cranial base who, was treated by a combination of aggressive surgical and medical therapy and subsequently had surgical repair of the oral defect. The purpose of this presentation is to draw attention to the clinical presentation and pathogenesis of rhinocerebral mucormycosis and to emphasise the need for high index of suspicion in its diagnosis and management. PMID- 12638837 TI - Improving the quality of reports of randomised controlled trials: the CONSORT statement. PMID- 12638838 TI - The changing spectrum of type 2 diabetes mellitus. PMID- 12638839 TI - Nephropathy in patients with recently diagnosed type 2 diabetes mellitus in black Africans. AB - BACKGROUND: Albuminuria is long recognised as a sign of renal disease in diabetes. In type 1 diabetes, renal disease occurs after a longer duration of diabetic state. In type 2 diabetes, it is more variable. OBJECTIVE: To determine the prevalence and any risk factors of albuminuria in short-term (< or = 2 yrs) type 2 diabetes. DESIGN: Cross sectional, descriptive study. Microalbuminuria was assessed using micro II strips. SETTING: Outpatient diabetic clinic at Kenyatta National Hospital, Nairobi. SUBJECTS: Patients who were newly diagnosed or had had type 2 diabetes for two years or less. MAIN OUTCOME MEASURES: Microalbuminuria, lipids, glycated haemoglobin, fasting blood glucose and blood pressure. RESULTS: One hundred and thirty nine patients who had type 2 diabetes mellitus for < or = 2 yrs were seen, but only 100 patients were included in the study over a six month period. Their mean (SD) age was 53.7 (9.3) years. Mean (SD) duration of diabetes was 10.3 (7.5) months. Fifty per cent of the study patients were hypertensive. Only 48% had HbA1c < 8% while 36% had HbA1c > 9%. The lipid profile of total, LDL-HDL-cholesterol and triglycerides were predominantly within normal limits. Twenty six per cent were established to have albuminuria of which one patient had macroalbuminuria. Blood pressure, glycated haemoglobin and lipid parameters were not significantly different from patients without albuminuria CONCLUSION: Albuminuria occurred in a significant proportion of patients with short term type 2 diabetes. Comparable to studies done elsewhere on short-term type 2 diabetes, albuminuria is both a sign of nephropathy and a cardiovascular risk factor. It should be looked for in all patients with type 2 diabetes attending this clinic, even at diagnosis. PMID- 12638840 TI - Platelet functions in patients with meningococcal meningitis at the Kenyatta National Hospital, Nairobi. AB - OBJECTIVE: To determine platelet abnormalities in patients with menigococcal meningitis. DESIGN: Case control study. SUBJECTS: Fifty seven cases of mennigococcal meningitis based on a cerebrospinal fluid gram stain for gram negative diplococcus or positive culture were recruited. Fifty-seven controls matched for age and sex were also recruited. The following platelet functions tests were performed; platelet counts, platelet adhesiveness, platelet aggregation and clot retraction. RESULTS: Fifty seven patients (41 males and 16 females) with meningococcal meningitis were studied. Their mean age was 25.5 +/- 8.32 years with a range of 15 to 45 years. Five patients had purpura, four peripheral gangrene, eight conjunctival haemorrhages and one was in shock. There was a statistical significant difference in the platelet aggregation and clot retraction between the patients and controls at p-values of 0.0001 and 0.0002 respectively. There was no significant difference in the platelet count and adhesiveness between the patients and the controls at a p-value of 0.203 and 0.22 respectively. No association was found between the platelet functions and the clinical presentations. CONCLUSION: Patients with meningococcal meningitis have abnormalities in the platelet functions mainly in aggregation and adhesiveness. PMID- 12638841 TI - Medical students perception of problem topics in anatomy. AB - OBJECTIVE: To examine the existence of perceived problem topics/concepts in anatomy, possible reasons for the problems and also student suggestions for alleviating the problems. DESIGN: An open-ended questionnaire. SETTING: School of Anatomical Sciences, University of the Witwatersrand, Johannesburg, South Africa. SUBJECTS: Two hundred and fifty nine second year medical students. MAIN OUTCOME MEASURES: The questionnaire included identification of problem areas, reasons for problems and possible solutions. RESULTS: The survey revealed a wide range of problem topics, with the majority being experienced in gross anatomy followed by histology and embryology. It was clear that the students experienced difficulties with 3-dimensional conceptualization and the visualisation of structures. CONCLUSION: A variety of teaching aids may help in addressing this problem and encourage spatially oriented thinking, leading ultimately to an integrated and in depth understanding of anatomy. PMID- 12638842 TI - Multiple drug resistance in urinary pathogens at Gondar College of Medical Sciences Hospital, Ethiopia. AB - OBJECTIVE: To determine multiple drug resistance and its associated factors of urinary pathogens. DESIGN: Cross-sectional study. SETTING: Gondar College of Medical Sciences teaching and Referral Hospital, Northwest Ethiopia, between January and October 2000. SUBJECTS AND METHODS: Mid stream urine samples from 420 subjects were studied by quantitative culture method. Designed Questionnaires were used for data collection on the previous use of antimicrobials, catheterisation and hospitalisation. MAIN OUTCOME MEASURES: Rates of multiple drug resistance and the associated factors. RESULTS: Multiple drug resistance was common in the isolates tested against ten antibiotics showing more than 68% of the isolates being resistant to two or more antimicrobials. Significant variables associated with this were found to be urinary catheterisation, hospitalisation and previous use of antibiotics for urinary tract infection. CONCLUSION: The rate of multiple drug resistance was very high in this study. Probable contributing factors were found to be previous antibiotic exposure, urinary catheterisation and hospitalisation. Reduction of hospital stays and catheterisation, aseptic care of catheterised patients and selective use of antibiotics and strict follow up of hospital disease controls are recommended. PMID- 12638844 TI - Role of Campylobacter jejuni/coli in diarrhoea in Ile-Ife, Nigeria. AB - BACKGROUND: Campylobacter jejuni/coli are well established causative agents of diarrhoea. In Nigeria, gastroenteritis due to C. jejuni was first reported in northern part of the country in 1981 and the South-Western part in Ile-Ife in 1983. OBJECTIVE: To re-examine the role of C. jejuni as an agent of diarrhoea after more than a decade at Ile-Ife, Nigeria and to determine the biological characteristics of local strains. DESIGN: A prospective case control study. SETTING: Obafemi Awolowo University Teaching Hospitals Complex (OAUTHC) Ile-Ife, Nigeria. SUBJECTS: Three hundred and three children with diarrhoea and 100 children with other medical conditions other than diarrhoea were randomly selected. MAIN OUTCOME MEASURES: Isolation of C. jejuni/coli from stool samples collected from the patients and controls. Biological characteristics of the isolates. RESULTS: Fifty eight (19.1%) and 6% of the patients and controls, had the organism respectively. Campylobacter coli accounts for 53.3% of isolates. All the isolates were susceptible to erythromycin and there was no evidence of beta lactamase production. CONCLUSION: Campylobacter jejuni is an important diarrhoea agent in our environment and should be considered strongly in children with diarrhoea. Complete characterization of local, strains is necessary. PMID- 12638843 TI - Quantitative analysis of gait pattern in hemiparetic patients. AB - OBJECTIVE: To characterise gait pattern in hemiparetic patients quantitatively using clinical footprint method. DESIGN: A case control study. SUBJECTS: Sixteen hemiparetic patients (12 males and 4 females) aged 16 to 64 years who attended neurological clinic at Queen Elizabeth Central Hospital, Blantyre, Malawi. MAIN OUTCOME MEASURES: Stride length, step width, foot rotation angle measured using footprint method. RESULTS: The difference in mean values of the three variables of gait between affected and unaffected sides in hemiparetic patients was not significant. However, strides were significantly longer in controls than in patients (P<0.001) while patients had significantly broader steps (P<0.02). In the controls, foot was rotated externally in 87.7% of steps and internally in 14.3% of steps. In the hemiparetic patients, intoeing pattern was observed in 41.3% of steps. The difference in variability of stride length and foot angle between affected and non-affected sides of patients was not significant. Stride to-stride variability in stride length in patients was 1.6 times higher than in controls. Variability of step width and foot progression angle was 1.4 and 1.6 times higher in patients than in controls. CONCLUSION: Footprint method provides fast and inexpensive tool for clinical gait analysis and is suitable for evaluation of hemiparetic patients. Our findings suggest that areas of emphasis for physical therapy of hemiparetic patients should include increasing stride length and decreasing step width and internal rotation of foot, particularly on the affected side. PMID- 12638845 TI - Nutrition status of children in Kasese district at the Uganda-Congo border. AB - OBJECTIVE: To establish the magnitude and risk factors for malnutrition in Kasese district at the Uganda-Congo border. DESIGN: Cross sectional nutrition survey. METHODS: Thirty clusters were selected. The height, weight, and mid upper arm circumference of at least 31 children per cluster were measured. Data on food frequency, prevalence of cough, fever and diarrhoea was also collected. RESULTS: Half of the 932 children (49.8%) were stunted, and 21.9% were severely stunted. While 17.4% of the children were under weight, 1.29% were wasted and 3.7% had MUAC <12.5 cm. Risk factors for stunting included not consuming milk, fathers' low formal education, having no immunisation card, and not breast feeding among 12-23 months old. Risk factors for underweight included fathers' and mothers' low education level, consumption of legumes, and fever or diarrhoea two weeks before the survey. For wasting the risk factors were consumption of vegetables, legumes or starchy foods. Those not consuming high-energy or animal foods including milk were likely to be severely wasted as were those not breast feeding in the 12-23 months age group. Fever or a diarrhoea episode two weeks preceding the survey and living in a female-headed household, were also risk factors for severe wasting. CONCLUSION: Attention to education, poverty alleviation, appropriate feeding practices, prevention and treatment of infections might assist in combating malnutrition in this district. PMID- 12638846 TI - Nosocomial infections in developing countries: cost effective control and prevention. AB - OBJECTIVES: To review the efficient and cost-effective preventive, control and surveillance measures that could be employed against nosocomial infections in developing countries. DATA SOURCES: Literature search on compact disk-read only memory (CD-ROM), Medline and Internet, using the key words: nosocomial infection, prevention and control, use of antibiotics and use of computers. Some articles were manually reviewed. STUDY SELECTION: Relevant studies or articles on nosocomial infections in developing and developed countries were included in the review. DATA EXTRACTION: From individual studies or articles. DATA SYNTHESIS: Information on nosocomial infections from developing and developed countries with some emphasis on Kenya is synchronized under the headings; introduction, historical background of nosocomial infections. Current situation of nosocomial infections and predisposing factors, nosocomial infections and antimicrobial resistance, consequences of nosocomial infections, hospital infection control programme and use of computers in nosocomial infection surveillance, and the cost benefit of infection prevention and control programme. CONCLUSION: Nosocomial infections may be contained more effectively by having an infection prevention and control programme. Computer-assisted epidemiological surveillance appears to be the most important aspect of monitoring infection control programmes, and to identify changes in risk factors that can increase the infection rate. Even minimally, effective infection control programmes are cost-effective. For the war against nosocomial infections to be won, the whole exercise should be handled as a global project with significant inputs from developing countries. PMID- 12638847 TI - Soil transmitted nematodes in children in Buea Health District of Cameroon. AB - OBJECTIVE: To investigate the prevalence, intensity of infection and control of soil-transmitted nematodes in primary school pupils of urban, suburban and rural localities. SETTING: Buea Health District. DESIGN: A cross section experimental study. SUBJECTS: Three hundred and forty primary school children. INTERVENTION: Mebendazole treatment complemented with health education. RESULTS: The overall prevalence rate of soil-transmitted nematodes in the Buea Health District was 59.1%. The infection rate was significantly different in the urban, suburban and rural localities of the health district (P<0.01), with the rural area having the highest rate of infection. Ascaris lumbricoides was the most predominant parasite. The intensity of infection with the various helminth species was not significantly different between children of urban, suburban and rural localities. Mebendazole treatment significantly reduced the intensities of infection with Ascaris and Trichuris (P<0.05). The treatment, however, was ineffective against hookworm infection. Re-infection rate with Ascaris and Trichuris was slower in children who additionally received health education than in those who received only anthelminthic treatment. CONCLUSION: Ascaris lumbricoides, Trichuris trichiura and hookworms are the predominant nematodes reported in children of urban, suburban and rural localities in the health district. Health education is advocated as a complement to chemotherapy in the control of soil-transmitted nematodes. PMID- 12638849 TI - Wandering spleen presenting as a right hypochondrial mass and intestinal obstruction. AB - This is a case report of a 23 year old multiparous woman who presented with intestinal obstruction and a right hypochondrial mass. Laparatomy revealed an infarcted 1.4 Kg spleen in the right lumbar region compressing the ascending colon. There was also ileal volvulus around the splenic pedicle. This is probably the first documented case of wandering spleen in the right hypochondrium, presenting as right large bowel obstruction, to be reported in our region. Wandering spleen is a rare condition, often asymptomatic, but may present as an acute abdomen. Pre-operative diagnosis is difficult and rarely made. Laboratory tests are seldom useful, but imaging studies do assist. Up to 1971 only 350 cases had been reported in the western literature. Review of English literature from 1900 to 1991 reported only 51 cases in children. In our region 11 cases were reported in Uganda between 1968 and 1971. No other literature is available from our region. Clinical presentation, aetiology, investigation, and management of wandering spleen is discussed. PMID- 12638850 TI - Rideal lecture. Universal features of the fluid to solid transition for attractive colloidal particles. AB - Attractive colloidal particles can exhibit a fluid to solid phase transition if the magnitude of the attractive interaction is sufficiently large, if the volume fraction is sufficiently high, and if the applied stress is sufficiently small. The nature of this fluid to solid transition is similar for many different colloid systems, and for many different forms of interaction. The jamming phase transition captures the common features of these fluid to solid translations, by unifying the behavior as a function of the particle volume fraction, the energy of interparticle attractions, and the applied stress. This paper describes the applicability of the jamming state diagram, and highlights those regions where the fluid to solid transition is still poorly understood. It also presents new data for gelation of colloidal particles with an attractive depletion interaction, providing more insight into the origin of the fluid to solid transition. PMID- 12638848 TI - Levels of malondialdehyde, glutathione and ascorbic acid in idiopathic thrombocytopaenic purpura. AB - BACKGROUND: Idiopathic thrombocytopaenic purpura (ITP) is an autoimmune disorder, which causes an acute or chronic thrombocytopenia, and may result in potentially life-threatening haemorrhage. Oxidative damage may be involved in the pathogenesis of autoimmune diseases. Antibodies to bind to membrane lipids and platelet destruction may play a role on lipid peroxidation in ITP. OBJECTIVES: To investigate the posible role of lipid peroxidation and antioxidants in patients with ITP. DESIGN: The levels of plasma and erythrocyte malondialdehyde (MDA), erythrocyte glutathione and ascorbic acid were analysed in patients with ITP. METHODS. The MDA levels were performed according to the method of Bidlack WR. Plasma MDA, erythrocyte glutathione and ascorbic acid levels were carried out according to the methods of Ohkawa H, Beutler E and Bauer JD, respectively. RESULTS: The erythrocyte and plasma MDA levels in patients with ITP were found to be 9.52+/-4.65, 3.03+/-1.44 (p<0.001) and in control group were found to be 2.49+/-0.57, 1.03+/-0.28 nmol/ml (p<0.001), respectively. Erythrocyte glutathione was found to be 3.71+/-0.82, 6.26+/-0.66 micromol/gr Hb (p<0.001). Ascorbic acid levels of these groups were 1.09+/-0.25, 1.70+/-0.33 mg/dl (p<0.001). CONCLUSION: The oxidative damage is involved in the pathogenesis of ITP. In patients with ITP, the platelet destruction and bleeding may play significant role on elevation of lipid peroxidation and reduction of antioxidant capacity. Further studies on oxidant and antioxidant status of ITP are also needed to confirm these results. PMID- 12638851 TI - The nature of the colloidal 'glass' transition. AB - The dynamically arrested state of matter is discussed in the context of athermal systems, such as the hard sphere colloidal arrest. We believe that the singular dynamical behaviour near arrest expressed, for example, in how the diffusion constant vanishes may be 'universal', in a sense to be discussed in the paper. Based on this we argue the merits of studying the problem with simple lattice models. This, by analogy with the the critical point of the Ising model, should lead us to clarify the questions, and begin the program of establishing the degree of universality to be expected. We deal only with 'ideal' athermal dynamical arrest transitions, such as those found for hard sphere systems. However, it is argued that dynamically available volume (DAV) is the relevant order parameter of the transition, and that universal mechanisms may be well expressed in terms of DAV. For simple lattice models we give examples of simple laws that emerge near the dynamical arrest, emphasising the idea of a near-ideal gas of 'holes', interacting to give the power law diffusion constant scaling near the arrest. We also seek to open the discussion of the possibility of an underlying weak coupling theory of the dynamical arrest transition, based on DAV. PMID- 12638852 TI - Real-space fluorescence recovery after photo-bleaching of concentrated suspensions of hard colloidal spheres. AB - In concentrated suspensions of fluorescent colloidal hard spheres (close to and above the glass transition density), we bleached part of the system in cube shaped regions using high intensity laser light. Recovery of these bleached cubes was followed in real space using confocal scanning laser microscopy (CSLM). This method provides mean squared particle displacements up to timescales that are three orders of magnitude beyond those available by present experimental techniques. We show that, above the (hard sphere) glass transition density, particles move over distances of the order of their own diameter on timescales of 10(6) to 10(8) Brownian times. Moreover, the mean squared displacement, (x2), shows powerlaw behavior over seven time (tau) decades: (x2) alpha tau(0.30 +/- 0.05). This behavior is different from earlier observations by dynamic light scattering. It is argued that these differences are caused by gravity effects, as the only difference between the systems is the buoyant mass of the colloids. PMID- 12638853 TI - Ergodic and non-ergodic phase transitions in globular protein suspensions. AB - The equilibrium and nonequilibrium phase behavior of a protein suspension is investigated as a function of strength of interparticle attraction and protein concentration. The equilibrium phase behavior suggests that the range of the particle attractions is a small fraction of their diameter. At volume fractions and strengths of attractions smaller than those characterizing the spinodal of a metastable fluid-fluid transition, the suspensions gel before they crystallize. At higher volume fractions and lower strengths of attraction, gels are formed first from which crystals nucleate over a period of time. However, at higher strength of attraction, crystals form first at lower volume fractions while gels are observed at higher volume fractions. We hypothesize that this behavior results from the competition between the rates of gelation and crystal nucleation. The location of the gel line is well predicted by mode coupling theories adapted for low volume fractions and square well fluids. At low strength of attraction, the gel line occurs at small supersaturations suggesting gels will be seen before nucleation. However, at higher strengths of attraction the gel line occurs at greatly increased supersaturations such that crystal nucleation can occur before the gel line is crossed. That mode coupling theories should predict gelation when crystallization does not intervene is tested by investigating the gel dynamics. PMID- 12638854 TI - The effect of free polymer on the interactions and the glass transition dynamics of microgel colloids. AB - By studying the phase behaviour and the short range order of polystyrene microgel colloids in a good solvent we demonstrated that these particles can be considered as hard spheres or nearly hard spheres if the internal cross link density is made sufficiently high. A binary mixture of 1:50 crosslinked (1 crosslink per 50 monomers) particles with a size ratio R(small)/R(large) = 0.83 and a number ratio N(small)/N(large) = 2.7 was characterized with respect to the short range order and glass transition dynamics. We found that this mixture can be mapped onto a one-component hard sphere system with an effective polydispersity of 11%, even though for the particle interactions an inverse power potential u(r) alpha r(-35) was established. The only significant difference observed was a shift of the glass transition to a higher volume fraction of 0.595. Thus, the binary mixture was used as a non-crystallizing reference system to study the effect of free polymer on interactions and glass transition dynamics. Addition of linear polystyrene with a size ratio delta = R(g,polymer)/R(colloid) = 0.054 induced short-ranged depletion attractions between microgel particles. In the case of a glassy sample already a rather small amount of polymer leads to melting of the glass and a re-entrant glass transition is observed at high polymer content. From the difference of the line shapes of the density autocorrelation functions on approaching the different transition lines. measured over the full dynamic range, a change of the freezing mechanism from a packing-driven glass transition at low polymer content to a bonding-driven glass transition at high polymer content was deduced. PMID- 12638855 TI - Phase transitions and gelation of silica-polystyrene mixtures in benzene. AB - We have performed experiments on mixtures of sterically stabilised silica particles and large poly(styrene) polymers in benzene. Using this model system, we were able to independently vary short and long ranged attractive interactions by changing the temperature and/or the polymer concentration. We show that the addition of long ranged attractions to particles with initially only short ranged attractions leads to the appearance of a gas-liquid phase transition. Our results are in accordance with earlier theoretical predictions. PMID- 12638856 TI - Nonequilibrium pattern formation in strongly interacting driven colloids. AB - Dynamical instabilities are discussed for strongly interacting colloidal suspensions which are driven into nonequilibrium by an external field in the limit where hydrodynamic interactions can be neglected. Brownian dynamics computer simulations indicate that stripe-like patterns of particles driven alike are spontaneously formed if the external drive exceeds a critical strength. Recent previous studies of stripe formation obtained for symmetric equimolar mixtures in the steady state are reviewed. These results are then extended in two directions: first, we show that stripe-like segregation occurs also in asymmetric mixtures and observe an additional compression/expansion effect in the stripes composed of the small/large particles. Second, we study the relaxation into the stripe-patterned steady state starting from a uniform demixed state and show that different transient processes such as jamming, anisotropic coarsening and void formation are relevant on the route into the stratified steady state. PMID- 12638857 TI - Colloidal epitaxy: playing with the boundary conditions of colloidal crystallization. AB - We have studied, with quantitative confocal microscopy, epitaxial colloidal crystal growth of particles interacting with an almost hard-sphere (HS) potential in a gravitational field and density matched colloids interacting with a long range (LR) repulsive potential with a body-centred cubic (BCC) equilibrium crystal phase. We show that in both cases it is possible to grow thick, stacking fault-free metastable crystals: close-packed crystals with any stacking sequence, including hexagonal close packed (HCP), for the HS particles and face-centred cubic (FCC) in the case of the LR colloids. In accordance with recent computer simulations done for HS particles it was found that the optimal lattice constant to grow HS HCP crystals was larger than that of equilibrium FCC crystals. In addition, because of the absence of gravity, pre-freezing could be observed for the particles with the LR potential on a template of charged lines. We also argue that the ability to manipulate colloids with highly focused light, optical traps or tweezers, will become an important tool in both the study of colloidal crystallization and in making new structures. We show how cheap 2D and 3D templates can be made with optical tweezers and demonstrate, in proof of principle experiments with core-shell colloids, how light fields can generate crystal nuclei and other structures in the bulk of concentrated dispersions and how the effect of these structures on the rest of a dispersion can be studied quantitatively in 3D. PMID- 12638858 TI - Self-assembly of a binary mixture of particles and diblock copolymers. AB - Using theoretical models, we undertake the first investigation into the synergy and rich phase behavior that emerges when binary particle mixtures are blended with microphase-separating copolymers. We isolate an example of spontaneous hierarchical self-assembly in such hybrid materials, where the system exhibits both nanoscopic ordering of the particles and macroscopic phase transformation in the copolymer matrix. Furthermore, the self-assembly is driven by entropic effects involving all the different components. The results reveal that entropy can be exploited to create highly ordered nanocomposites with potentially unique electronic and photonic properties. PMID- 12638859 TI - Sheared colloidal crystals in confined geometry: a real space study on stationary structures under shear. AB - We constructed an optical plate-plate shear cell suitable for the study of aqueous suspensions of charged colloidal spheres under low electrolyte concentrations (10(-6) M). The variable gap height was adjusted to 30 microm corresponding to 15-30 interparticle distances. The concentration of 300 microm diameter polystyrene spheres was chosen around 1 microm(-1) where previous studies had revealed the equilibrium structure to be fluid or body centred cubic. Under shear, layer structures of hexagonal symmetry form, often coexistent with a fluid phase. We used an adapted high resolution video microscope to perform a detailed study of the structure. The central ray of light was prevented from entering the objective and the images are constructed from higher order scattered light. The cover glass correction was adjusted in such a way that several layers of particles were visible simultaneously. The effective depth of sharpness was thus increased to several microm. From the 2D video frames we constructed time averaged 2D particle correlation diagrams (PCD) which correspond to the distribution functions for the projections of particle positions into the x y plane. They allow us to discriminate different layers and to analyse their structure and their relative position. Layer structures showed approximately hexagonal symmetry bat with an elongation in the flow direction. They were always observed to be wall based. Melting in the z-direction occurred before in-plane melting and with a different mechanism. The resulting salt concentration-shear rate phase diagram is discussed in comparison with results from computer simulation. PMID- 12638860 TI - Structure and dynamics of particle monolayers at a liquid-liquid interface subjected to shear flow. AB - The effect of shear flow on the structure and dynamics of monodisperse spherical polystyrene particles suspended at the interface between decane and water was observed. While undisturbed, the particles arrange themselves on a hexagonal lattice due to strong dipole-dipole repulsion resulting from ionizable sulfate groups on their surfaces. As the interface is subjected to shear flow, however, the lattice adopts a new semi-ordered, anisotropic state for which two distinct regimes are observed. At low particle concentrations or high shear rates, nearest neighbors in the lattice align in the flow direction and create strings of particles that slip past each other fairly readily. This results in a stretching of the overall structure and achievement of a steady state orientation in the system. In contrast, at high concentrations or low shear rates, the interparticle forces gain importance and tend to keep the particles more strongly in their lattice positions. As a result, domains within the lattice are forced to rotate, thus giving rise to movement of particles perpendicular to the flow direction. Thus a rotation, in addition to stretching, of the structure is apparent in this case. PMID- 12638862 TI - The influence of surface forces on thin film drainage between a fluid drop and a flat solid. AB - An experiment is described in which a mica surface is driven towards a mercury drop immersed in aqueous electrolyte. Under appropriate conditions, hydrodynamic pressure in the aqueous film creates a classical dimple in the mercury drop. The use of optical interferometry and video recording to monitor the shape of the drop and the thickness of the aqueous film with sub-nanometre resolution yields a high density of precise data showing the formation and evolution of the dimple as the film drains. Variation of electrical potential applied to the mercury phase allows control of the surface forces acting between the drop and the mica surface, so that the effect of surface forces on the film drainage process is highlighted. It is found that the film thickness at the centre of the dimple and the lateral extent of the dimple are not significantly affected by surface forces. On the other hand, the minimum film thickness at the edge of the dimple is sensitive even to weak surface forces. Since this minimum film thickness is a major determinant of the film drainage rate, it is shown that surface forces have an important effect on the overall drainage process. PMID- 12638861 TI - Shear-banding and microstructure of colloids in shear flow. AB - We shall discuss the following phenomena found in various colloidal systems in shear flow. We recently observed shear-banding in suspensions of fd-virus in a cylindrical shear cell. Small angle light scattering experiments revealed that the shear-banding transition is preceded by a relatively fast process (minutes) of nematic-to-paranematic phase separation during which inhomogeneities on the micrometer length scale are formed. After the formation of these inhomogeneities, a slow (hours) appearence of shear-bands is observed, which have a height of a few mm. In the stationary state it is found, by means of polarization microscopy, that inhomogeneities exist within the bands. Small angle, time resolved light scattering experiments on near-critical microstructural order in a mixture of colloidal spheres and free polymer under stationary shear flow are discussed. The unexpected distortion of microstructure in directions perpendicular to the flow direction is quantitatively explained by extending an already existing theory, to include shear-induced short-ranged microstructural distortion. In colloidal systems consisting of "hairy colloids", where a spherical core is decorated with relatively long polymers, shear-induced polymer brush deformation might be important for its structural and rheological behavior. Preliminary neutron scattering and rheology experiments are performed to study polymer brush deformation in (semi-) dilute suspensions. PMID- 12638863 TI - Interactions between sickle hemoglobin fibers. AB - We report on observations of "zippering" that occurs when two sickle hemoglobin fibers come together side by side. A transient Y-shaped object is formed which "zips " closed. We have been able to show how the strength of the interactions that drive this may be estimated by studying the frustrated structures sometimes formed between several fibers. Our measurements, when combined with mechanical constants determined by an analysis of bending fluctuations, allow us to make the first estimate of the magnitude of these interactions, of the order of 7kBT microm(-1). Hemoglobin volume fractions of tens of %, lead to significant depletion forces. We estimate the magnitude of both the depletion and Van der Waals forces between pairs of single fibers. We study how these are effected by the helical nature of the fibers and renormalised by bending fluctuations, calculations that could have wider applications beyond sickle hemoglobin fibers. Our theoretical analysis of single fibers is in encouraging, although not fully quantitative, agreement with our measurements. We conclude that the physics and rheology of the hemoglobin gel, as well as the pathology of sickle cell anemia itself, may be influenced by depletion interactions. PMID- 12638865 TI - How are colloidal suspensions that age rejuvenated by strain application? AB - We present here a microscopic study of the effect of shear on a dense purely repulsive colloidal suspension. We use multispeckle diffusing wave spectroscopy to monitor the transient motions of colloidal particles after being submitted to an oscillatory strain. This technique proves efficient to record the time evolution of the distribution of relaxation times. After a high oscillatory shear, we show that this distribution displays full aging behavior. Conversely, when a moderate shear is applied the distribution is modified in a non-trivial way. Whereas high shear is able to erase all the sample history and rejuvenate it, a moderate shear helps it to age. We call this phenomenon overaging. We demonstrate that overaging can be understood if the complete shape of the relaxation time distribution is taken into account. We finally report how the soft glassy rheology model accounts for this effect. PMID- 12638866 TI - Schematic models for dynamic yielding of sheared colloidal glasses. AB - The nonlinear rheological properties of dense suspensions are discussed within simplified models, suggested by a recent first principles approach to the model of Brownian particles in a constant-velocity-gradient solvent flow. Shear thinning of colloidal fluids and dynamical yielding of colloidal glasses arise from a competition between a slowing down of structural relaxation, because of particle interactions, and enhanced decorrelation of fluctuations, caused by the shear advection of density fluctuations. A mode coupling approach is developed to explore the shear-induced suppression of particle caging and the resulting speed up of the structural relaxation. PMID- 12638864 TI - Universal non-diffusive slow dynamics in aging soft matter. AB - We use conventional and multispeckle dynamic light scattering to investigate the dynamics of a wide variety of jammed soft materials, including colloidal gels, concentrated emulsions, and concentrated surfactant phases. For all systems, the dynamic structure factor f(q,t) exhibits a two-step decay. The initial decay is due to the thermally activated diffusive motion of the scatterers, as indicated by the q(-2) dependence of the characteristic relaxation time, where q is the scattering vector. However, due to the constrained motion of the scatterers in jammed systems. the dynamics are arrested and the initial decay terminates in a plateau. Surprisingly, we find that a final, ultraslow decay leads to the complete relaxation of f(q,t), indicative of rearrangements on length scales as large as several microns or tens of microns. Remarkably, for all systems the same very peculiar form is found for the final relaxation of the dynamic structure factor: f(q,t) approximately exp[-(t/tau s)p], with p approximately equal to 1.5 and tau s approximately q(-1), thus suggesting the generality of this behavior. Additionally, for all samples the final relaxation slows down with age. although the aging behavior is found to be sample dependent. We propose that the unusual ultraslow dynamics are due to the relaxation of internal stresses, built into the sample at the jamming transition, and present simple scaling arguments that support this hypothesis. PMID- 12638867 TI - Yielding and flow of colloidal glasses. AB - We investigate the yielding and flow of hard-sphere colloidal glasses by combining rheological measurements with the technique of light scattering echo. The polymethylmethacrylate particles used are sufficiently polydisperse that crystallization is suppressed. Creep and recovery measurements show that the glasses can tolerate surprisingly large strains, up to at least 15%, before yielding irreversibly. We attribute this behaviour to 'cage elasticity', the ability of a particle and its cage of neighbours to retain their identity under quite large distortion. Results from light scattering echo, which measures the extent of irreversible particle rearrangement under oscillatory shear, support the notion of cage elasticity. In the lower concentration glasses we find that particle trajectories are partly reversible under strains which significantly exceed the yield strain. PMID- 12638868 TI - Colloidal dynamics in polymer solutions: optical two-point microrheology measurements. AB - We present an extension of the two-point optical microrheology technique introduced by Crocker et al. [J. C. Crocker, M. T. Valentine, E. R. Weeks, T. Gisler, P. D. Kaplan, A. G. Yodh and D. A.Weitz, Phys. Rev. Lett., 2000, 85, 888] to high frequencies. The correlated fluctuations of two probe spheres held by a pair of optical tweezers within a viscoelastic medium are determined using optical interferometry. A theoretical model is developed to yield the frequency dependent one- and two-particle response functions from the correlated motion. We demonstrate the validity of this method by determining the one- and two-particle correlations in a semi-dilute solution of polystyrene in decalin. We find that the ratio of the one- and two-particle response functions is anomalous which we interpret as evidence for a slip boundary condition caused by depletion of polymer from the surface of the particle. PMID- 12638869 TI - On the validity of Stokes-Einstein-Debye relations for rotational diffusion in colloidal suspensions. AB - According to the Stokes-Einstein-Debye (SED) relation, the rotational diffusion coefficient of a colloidal tracer sphere scales with the inverse of the solvent viscosity. Here we investigate the generalization of the SED relation to tracer diffusion in suspensions of neutral and charged colloidal host spheres. Rotational diffusion coefficients are measured with dynamic light scattering and phosphorescence spectroscopy, and calculated including two- and three-particle hydrodynamic interactions. We find that rotational tracer diffusion is always faster than predicted by the SED relation, except for large tracer/host size ratios lambda. In the case of neutral particles this observation is rationalized by introducing an apparent lambda-dependent slip boundary coefficient. For charged spheres at low ionic strength, large deviations from SED scaling are found due to the strongly hindered host sphere dynamics. Finally, we present some first experiments on tracer sphere diffusion in suspensions of host rods, showing that hydrodynamic hindrance by rods is much stronger than by spheres. We conclude by pointing to some interesting unresolved issues for future research. PMID- 12638870 TI - Colloid flow during thickening--a particle level understanding for core-shell particles. AB - The flow curve of sheared concentrated colloids can show a region of shear thickening. The underlying mechanisms involved in this effect has long been an issue. Recently the author and co-workers have used Stokesian dynamics approximated for high concentrations to simulate models of polymer coated particles in the thickening regime. This work continues the search for a particle level of understanding these systems in the thickening regime. Previous work found that shear thickening is associated with the formation of a network of contacts between polymer coats. Previous work by the author has examined the fabric (geometry) and texture (density distribution) of the contact network. However despite this many-body effect, it was also found that a very simple mean field type argument at the level of a pair of particles in contact relates the particle interaction laws to the thickening part of the flow curve up to an unknown factor. In the argument this factor is determined by aspects of the destruction of particle contacts. In this paper several new results are reported. The first concerns the definition of the network and its coordination. The paper contrasts systems with different thickness of polymer coats and gives tentative evidence that the network formed sits close to what is known as the iso-static coordination. The second set of issues concerns the analysis of the lifetimes of particle contacts--in effect an examination of the unknown factor and assumptions of the pair argument. A surprising result is that some particle pairs have relatively long lifetimes. Finally, data is reported for the stress fluctuations in the thickening regime. PMID- 12638871 TI - Fundamentals of aggregation in concentrated dispersions: fiber-optic quasielastic light scattering and linear viscoelastic measurements. AB - Fiber-optic quasielastic light scattering and oscillatory shear rheology are employed to monitor the structure formation in concentrated, aqueous dispersions of model, electrostatically stabilized, polymer colloids. A regime of fractal scaling is observed in the vicinity or the gel point, as signified by a power law decay in the autocorrelation function with delay time and a power law dependence of the shear moduli on frequency. The power law exponents and the gel times extracted from the two techniques are compared for the first time on the same dispersion. The details of structure formation and aggregation kinetics in these concentrated dispersions are compared to the universal behavior of DLCA and RLCA aggregation observed at dilute concentrations in these and other dispersions, as well as to polymer gelation. PMID- 12638873 TI - Aberrations of diffracted wave fields: distortion. AB - Near-field diffraction patterns are merely aberrated Fraunhofer diffraction patterns. These aberrations, inherent to the diffraction process, provide insight and understanding into wide-angle diffraction phenomena. Nonparaxial patterns of diffracted orders produced by a laser beam passing through a grating and projected upon a plane screen exhibit severe distortion (W311). This distortion is an artifact of the configuration chosen to observe diffraction patterns. Grating behavior expressed in terms of the direction cosines of the propagation vectors of the incident and diffracted orders exhibits no distortion. Use of a simple direction cosine diagram provides an elegant way to deal with nonparaxial diffraction patterns, particularly when large obliquely incident beams produce conical diffraction. PMID- 12638874 TI - Transmission-reflection analysis for localization of temporally successive multipoint perturbations in a distributed fiber-optic loss sensor based on Rayleigh backscattering. AB - A novel method is presented for the localization of multipoint loss-inducing perturbations in a distributed fiber-optic sensor. The proposed simple technique is based on measurement of the transmitted and the Rayleigh-backscattered powers of an unmodulated light launched into a sensing fiber. The positions of consecutive perturbations are determined by measuring the slopes of the dependence of normalized Rayleigh-backscattering power versus the square of normalized transmitted power. It is shown that these slopes uniquely depend on the positions of the disturbances along the test fiber. The method allows localization of any number of the perturbations that appear one after another at different positions along the test fiber without ambiguity. Good agreement is obtained between calculated and experimentally measured slopes for a loss that was consecutively induced near the source and remote ends of 2.844-km-long fiber. PMID- 12638872 TI - Particle dynamics in concentrated colloidal suspensions. AB - We have studied the dynamical and structural properties of dense charge stabilized colloidal suspensions by a combination of small-angle neutron scattering (SANS), 3D dynamic light scattering (3DDLS) and diffusing wave spectroscopy (DWS). SANS and 3DDLS extend static and dynamic light scattering techniques to the regime of opaque samples while DWS additionally provides information on short length scales, typically from 1-50 nm. This offers an increased range of accessible length and time scales perfectly suited for the (non-invasive) investigation of highly concentrated suspensions. Different systems have been examined ranging from hard sphere like suspensions over strongly repulsive charged spheres to colloidal gels. We furthermore present an extended theoretical frame for DWS to characterize the internal dynamics of turbid gels made from nanosized colloidal particles. PMID- 12638875 TI - Measurement method for profiling the residual stress of an optical fiber: detailed analysis of off-focusing and beam-deflection effects. AB - The effects of off-focusing and beam deflection on polarimetric stress measurements of optical fibers are investigated. A simple method for reducing the distortion of the phase retardation caused by unwanted beam deflections in residual stress measurement is introduced. The method is examined numerically by ray-tracing techniques and experimentally by use of hollow silica fibers into which various index-matching liquids have been inserted. An autofocusing technique is introduced. The error in stress measurement reproducibility was determined to be less than 4%. We tested the absolute error in measured stress by applying incremental external tension and determined that it is less than 0.464 MPa. PMID- 12638876 TI - Analysis of optical elements with flat boundary surfaces. AB - Optical systems are currently evaluated by use of ray-tracing techniques to extract performance quantities such as aberration and spot size. To improve on the use of optical equations, we formulate various important optical functions using a 4 x 4 homogeneous transformation matrix to design and analyze skew rays that cross flat optical boundary surfaces. We address three important topics: (1) the direction of a reflected or refracted ray is determined according to Snell's law, (2) sensitivity analysis expresses differential changes of reflected or refracted rays in terms of differential changes of incident rays, and (3) aberration of polychromatic light is presented analytically. A solid-glass corner cube and a Pechan prism are used to demonstrate the validity of the developed methodology. PMID- 12638877 TI - Holographic interference filters for infrared communications. AB - We demonstrate that high-quality interference filters for the wavelength range 1300-1600 nm can be holographically fabricated in DuPont HRF-800X001 photopolymer material by use of visible laser illumination. We also summarize a chain-matrix technique, which we call thin-film decomposition, that is useful for modeling multilayer films with an arbitrary index profile n(z). We use the thin-film decomposition technique to create design curves that allow one to choose the proper exposure angle and film thickness with which to fabricate a holographic interference filter with a desired transmission efficiency and bandwidth at a particular wavelength. These curves are of general utility and are not confined to any particular holographic recording medium. Excellent agreement between theory and experiment is found. PMID- 12638878 TI - Simulation of the depolarization effect in porous silicon. AB - We describe a radiative transfer (RT) equation for the simulation of optical scattering effects in a nanostructured semiconductor for spectroscopic ellipsometry (SE). As an example, we chose porous silicon (PS), whose pores are considered to act as light scatterers. We examined the effects of pore radius, slab thickness, and incident angle. The volume scattering effect in the internal morphology of the PS generates incoherent light, leading to depolarization. By simulating the four Stokes parameters through the RT equation, we could theoretically assess the degree of polarization that is essential for SE measurements of some nanostructured semiconductors. PMID- 12638879 TI - Spectral polarization measurements by use of the grating division-of-amplitude photopolarimeter. AB - The grating division-of-amplitude photopolarimeter (G-DOAP) is an instrument that exploits the multiple-beam-splitting, polarizing, and dispersive properties of diffraction gratings for the time-resolved measurement of the complete state of polarization of collimated broadband incident light, as represented by the four Stokes parameters as a function of wavelength across the spectrum. It is a compact, high-speed sensor that has no moving parts and is simple to install and operate. These characteristics make the G-DOAP well suited for in situ spectroscopic ellipsometry (SE) applications for monitoring and controlling thin film processes. The design and performance of a prototype instrument are presented. Precise SE measurements, to +/-0.04 degrees in psi and +/-0.1 degrees in delta, are demonstrated in the 550-940-nm wavelength range. PMID- 12638880 TI - Detection of transient surface acoustic waves of nanometric amplitude with double pulsed TV holography. AB - We describe the detection of bursts of surface acoustic waves by a double-pulsed TV holography technique. We describe mathematically the long- and short-wave bursts in the output correlograms and validate theoretical results with experimental images. The use of short-wave bursts permits us to scan the surface and makes it easier to distinguish, for purposes of nondestructive testing, the disturbances produced by flaws. PMID- 12638881 TI - Laser direct writing polymeric single-mode waveguide devices with a rib structure. AB - A focused argon-ion laser beam is used on a spin-coated polymeric thin-film deposited upon a SiO2/Si substrate to polymerize the core for fabrication of Gaussian profile optical channel waveguides. A rib structure that allows only the fundamental mode to propagate even with its higher dimension and high-index contrast between the core and the cladding was fabricated. When the thickness of the core-index region outside the rib section decreases, the waveguide produces higher-order modes at the output. The waveguide reported here has cross-sectional dimensions and numerical apertures that match the single-mode fibers for efficient coupling. I used a mixture of two intermiscible acrylate monomers for the cladding and the core of the waveguides. The polymerization process and its dependent dwell time or scan speed and the laser power intensity are shown. I present the operational characteristics of directional couplers using a rib waveguide structure with a core-cladding index difference. PMID- 12638882 TI - Sensing and control in dual-recycling laser interferometer gravitational-wave detectors. AB - We introduce length-sensing and control schemes for the dual-recycled cavity enhanced Michelson interferometer configuration proposed for the Advanced Laser Interferometer Gravitational Wave Observatory (LIGO). We discuss the principles of this scheme and show methods that allow sensing and control signals to be derived. Experimental verification was carried out in three benchtop experiments that are introduced. We present the implications of the results from these experiments for Advanced LIGO and other future interferometric gravitational-wave detectors. PMID- 12638883 TI - Dual-recycled cavity-enhanced Michelson interferometer for gravitational-wave detection. AB - The baseline design for an Advanced Laser Interferometer Gravitational-Wave Observatory (Advanced LIGO) is a dual-recycled Michelson interferometer with cavities in each of the Michelson interferometer arms. We describe one possible length-sensing and control scheme for such a dual-recycled, cavity-enhanced Michelson interferometer. We discuss the principles of this scheme and derive the first-order sensing signals. We also present a successful experimental verification of our length-sensing system using a prototype tabletop interferometer. Our results demonstrate the robustness of the scheme against deviations from the idealized design. We also identify potential weaknesses and discuss possible improvements. These results as well as other benchtop experiments that we present form the basis for a sensing and control scheme for Advanced LIGO. PMID- 12638884 TI - Signal extraction and optical design for an advanced gravitational-wave interferometer. AB - We present an experimental demonstration of a locking and control scheme for an interferometer using a power-recycled resonant sideband extraction configuration and show that the measured response to mirror vibrations matches an optical model. We discuss some aspects of resonant sideband extraction that are relevant to gravitational-wave detection. PMID- 12638885 TI - Power-recycled Michelson interferometer with resonant sideband extraction. AB - We present a system for the control and signal extraction of a power-recycled Michelson interferometer with resonant sideband extraction. This control system is capable of locking four degrees of freedom to a fixed point while locking the signal cavity to an arbitrary detuned point. One of the strengths of this system is that it can quickly change the instrument's frequency response without disrupting continuous operation. We report on an experimental implementation of this control system on a benchtop prototype and present broadband measurements of the prototype's frequency response for a range of signal cavity detunings. PMID- 12638886 TI - Spectral interference Mirau microscope with an acousto-optic tunable filter for three-dimensional surface profilometry. AB - A nonmechanical scanning Mirau-type spectral interference microscope has been developed for the measurement of three-dimensional surface profiles of discontinuous objects. An acousto-optic tunable filter (AOTF) is used as a high resolution spectral filter, which scans the optical frequency of the broadband light emitted from a superluminescent diode. To generate spectral fringes that make full use of the limited coherence length of the filtered light we unbalanced the Mirau interferometric system by positioning the reference mirror nearly halfway between the top and the bottom of the step height. When the frequency of the broadband light source is scanned by an AOTF, the interference fringes move in opposite directions on the top and the bottom of the object. To uniquely determine the sign of the fringe movement over the large area of the object, we developed a three-dimensional Fourier-transform technique, and from the detected sign of the fringe movement and phase information, we determined the three dimensional step height. Experimental results of the measurement of 100-microm step height are presented. The main advantages of the proposed system are that it provides nonmechanical scanning and a large measurement range without ambiguity in the sign of the phase. PMID- 12638887 TI - Automatic element addition and deletion in lens optimization. AB - A mechanism is established for the automatic addition and deletion of optical elements during the course of lens optimization. Two lens-form parameters, quantifying the symmetry of the optical system and the optical-power distribution among the individual lens elements, are used as criteria in this automatic procedure. Design examples are provided that demonstrate the practicability of the scheme. PMID- 12638888 TI - Development of a laser-scattering-based probe for on-line measurement of surface roughness. AB - The design and properties of an optical probe for on-line measurement of surface roughness are discussed. Based on light scattering, a probe that consists of a laser diode, a measuring lens, and a linear photodiode array was designed to detect surface roughness, in which the light scattered from a test surface at a relatively large scattering angle phi (=28 degrees) can be collected to enhance measuring range and repeatability. A coaxial design that incorporates a dual laser probe and compressed air makes the proposed system insensitive to the position of the test surface and to surface conditions such as the presence of debris, vibration, and lubricants that result from machining. The results from measurements of several sets of specimens have demonstrated the feasibility of measuring surface roughness by using light scattering. On-line measurement on a diamond-turning lathe has shown that the proposed technique is stable and compact enough to be applicable to on-line measurement of surface roughness of an engineering surface. PMID- 12638889 TI - Use of information on the manufacture of samples for the optical characterization of multilayers through a global optimization. AB - We present a procedure for the optical characterization of thin-film stacks from spectrophotometric data. The procedure overcomes the intrinsic limitations arising in the numerical determination of many parameters from reflectance or transmittance spectra measurements. The key point is to use all the information available from the manufacturing process in a single global optimization process. The method is illustrated by a case study of solgel applications. PMID- 12638890 TI - Multilayer thin-film structures with high spatial dispersion. AB - We demonstrate how to design thin-film multilayer structures that separate multiple wavelength channels with a single stack by spatial dispersion, thus allowing compact manufacturable wavelength multiplexers and demultiplexers and possibly beam-steering or dispersion-control devices. We discuss four types of structure--periodic one-dimensional photonic crystal superprism structures, double-chirped structures exploiting wavelength-dependent penetration depth, coupled-cavity structures with dispersion that is due to stored energy, and numerically optimized nonperiodic structures utilizing a mixture of the other dispersion effects. We experimentally test the spatial dispersion of a 200-layer periodic structure and a 66-layer nonperiodic structure. Probably because of its greater design freedom, the nonperiodic structure can give both a linear shift with wavelength and a larger usable shift than the thicker periodic structure gives. PMID- 12638891 TI - Symmetrical periods in antireflective coatings for plastic optics. AB - Plastic optical parts require antireflective as well as hard coatings. A novel design concept for coating plastics combines both functions. Symmetrical three layer periods with a phase thickness of 3/2pi are arranged in a multilayer to achieve a step-down refractive-index profile. It is shown mathematically that the equivalent index of symmetrical periods can be lower than the lowest refractive index of a material used in the design, if the phase thickness of the symmetrical period is set equal to 3/2pi instead of the usual pi/2. The straightforward application of the concept to the design of antireflection coatings in general is demonstrated by example. PMID- 12638892 TI - Diffuse reflection of ceramics coated with dielectric thin films. AB - We have studied the diffuse reflection properties of ceramics in the presence of dielectric thin films on the surface. A simple optical model was proposed in which interference effects in a thin film were considered for light scattered out of a ceramic in various directions. Measurements were performed on angle-resolved reflection spectra of a thin-film-coated alumina ceramic in the case of normal incidence. They showed that the presence of the thin film on the ceramic's surface modified the angular distributions of scattered radiation from that of a bare ceramic, which suggested a way to tailor the scattering properties of a diffuse reflector as needed. PMID- 12638893 TI - Computational design of solar reflection and far-infrared transmission films for a variable emittance device. AB - A smart radiation device (SRD) that is a variable emittance radiator has been studied as a method of thermal control for spacecraft. The SRD consists of manganese oxide with a perovskite-type structure, and the total hemispherical emittance of the SRD changes considerably depending on temperature. Here we propose an optimal method of designing multilayer films for the SRD by using agenetic algorithm. The multilayer films reflect solar radiation and transmit far infrared radiation to maintain variation of the infrared optical properties of the SRD. PMID- 12638894 TI - Optimum pulse duration and radiant exposure for vascular laser therapy of dark port-wine skin: a theoretical study. AB - Laser therapy for cutaneous hypervascular malformations such as port-wine stain birthmarks is currently not feasible for dark-skinned individuals. We study the effects of pulse duration, radiant exposure, and cryogen spray cooling (CSC) on the thermal response of skin, using a Monte Carlo based optical-thermal model. Thermal injury to the epidermis decreases with increasing pulse duration during irradiation at a constant radiant exposure; however, maintaining vascular injury requires that the radiant exposure also increase. At short pulse durations, only a minimal increase in radiant exposure is necessary for a therapeutic effect to be achieved because thermal diffusion from the vessels is minimal. However, at longer pulse durations the radiant exposure must be greatly increased. There exists an optimum pulse duration at which minimal damage to the epidermis and significant injury within the targeted vasculature occur. For example, the model predicts optimum pulse durations of approximately 1.5, 6, and 20 ms for vessel diameters of 40, 80, and 120 microm, respectively. Optimization of laser pulse duration and radiant exposure in combination with CSC may offer a means to treat cutaneous lesions in dark-skinned individuals. PMID- 12638895 TI - Influence of cell shape and aggregate formation on the optical properties of flowing whole blood. AB - We studied the influence of shape and secondary, or intercellular, organization on the absorption and scattering properties of red blood cells to determine whether these properties are of any practical significance for optical evaluation of whole blood and its constituents. A series of measurements of transmittance and reflectance of light from bovine blood in a flow cuvette was conducted with a 650-900-nm integrating sphere at shear rates of 0-1600 s(-1), from which the influence of cell orientation, elongation, and aggregate formation on the absorption (mu(a)) and the reduced scattering (mu(s)') coefficients could be quantified. Aggregation was accompanied by a decrease of 4% in mu(s)' compared with the value in randomly oriented single cells. Increasing the degree of cell alignment and elongation as a result of increasing shear rate reduced mu(s)' by 6% and mu(a) by 3%, evaluated at a shear rate of 1600 s(-1). Comparison with T matrix computations for oblate- and prolate-shaped cells with corresponding elongation and orientation indicates that the optical properties of whole blood are determined by those of its individual cells, though influenced by a collective scattering factor that depends on the cell-to-cell organization. We demonstrate that cell morphological changes must be taken into consideration when one is conducting whole blood spectroscopy. PMID- 12638896 TI - Clinical and epidemiological aspects in the antiphospholipid syndrome. AB - The antiphospholipid syndrome (APS) is defined by the occurrence of venous and arterial thromboses, often multiple, and pregnancy morbidity (mainly, recurrent fetal losses and premature births), frequently accompanied by a moderate thrombocytopenia, in the presence of antiphospholipid antibodies (aPL), namely lupus anticoagulant (LA), anticardiolipin antibodies (aCL), or both. Other autoantibodies have also been detected in many patients with an APS, such as anti beta2 glycoprotein I (GPI), antimitochondrial (M5 type), antiendothelial cell, antiplatelet, antierythrocyte, and antinuclear antibodies. The APS can be found in patients having neither clinical nor laboratory evidence of another definable condition (primary APS) or it may be associated with other diseases. Systemic lupus erythematosus (SLE) is the disorder in which an APS is most commonly associated. Less frequently, aPL and, rarely, an APS may also be encountered in other groups of patients (Table 1) (1). PMID- 12638897 TI - Antiphospholipid antibodies: are they pro-atherogenic or an epiphenomenon of atherosclerosis? AB - Antiphospholipid antibodies are the hallmark of the antiphospholipid syndrome which is characterized by thrombosis. There are currently data supporting an association between these autoantibodies and atherosclerosis as well. Human studies suggest that anti-cardiolipin and anti-beta2-glycoprotein-I antibodies are elevated in patients having coronary artery disease compared with controls. Anti-cardiolipin antibodies are also associated with typical chest pain, significant coronary artery stenosis on angiography and prediction of myocardial infarction. Laboratory studies and murine models support the pro-atherogenic role of these autoantibodies, as they are involved in uptake of oxidized LDL into macrophages, and immunization of mice with them results in enhanced atherosclerosis. There is some evidence that high anti-beta2-glycoprotein-I antibodies can present a risk factor for atherosclerosis, but more epidemiological data are required in order to confirm whether the pro-atherogenic properties of anti-phospholipid antibodies signifies an independent risk factor for atherosclerosis and its complications. PMID- 12638898 TI - Atherogenic autoantigen: oxidized LDL complexes with beta2-glycoprotein I. AB - Beta2-Glycoprotein I (beta2-GPI) is a major antigen for antiphospholipid antibodies present in patients with antiphospholipid syndrome (APS). In 1997, we demonstrated that beta2-GPI specifically binds to Cu2+-oxidized low-density lipoprotein (oxLDL) and that the beta2-GPI-oxLDL complex is subsequently targeted by anti-beta2-GPI antibodies in vitro. Then ligands for beta2-GPI were purified from oxLDL and characterized as omega-carboxylated 7-ketocholesteryl esters, such as 7-ketocholesteryl-9-carboxynonanoate (oxLig-1) and 7-ketocholesteryl-12 carboxy (keto) dodecanoate (oxLig-2). These ligands mediate to form oxLDL-beta2 GPI complexes, and the complexes are taken up avidly by macrophages via anti beta2-GPI autoantibody-mediated phagocytosis. We recently demonstrated that appearance of autoantibodies against a complex of beta2-GPI and oxLig-1 are highly associated with a history of arterial thrombosis. Serum oxLDL-beta2-GPI complex and their IgG immune complexes are also risk factors arterial thrombosis in APS patients. There is increasing circumstantial evidence of autoimmune mechanism involving beta2-GPI and oxLDL in the atherogenesis in APS. PMID- 12638899 TI - Atherosclerosis, oxidative stress and auto-antibodies in systemic lupus erythematosus and primary antiphospholipid syndrome. AB - Overwhelming evidence spanning three decades has consistently shown that coronary artery disease is a major cause of morbidity and mortality in Systemic Lupus Erythematosus. Traditionally this was explained by abnormalities of the lipid profile induced by prolonged steroid treatment. Subsequently, antiphospholipid antibodies were presented as an additional cardiovascular risk factor. Recently, antibodies towards high-density lipoprotein and antiapolipoprotein A-I have been identified. These, together with anti-beta2 glycoprotein-1, interfere with the major antioxidant defence of patients with SLE and with primary antiphospholiqid syndrome exposing them to the atherogenic potential of enhanced oxidative stress. The present review discusses how the latter auto-antibodies, together with abnormalities of their target lipid auto-antigens, could enhance the risk of atherosclerosis in SLE and APS. PMID- 12638900 TI - Endothelium as a target for antiphospholipid antibodies. AB - Several, and not necessarily alternative, pathogenic mechanisms have been suggested to sustain the thrombophilic diathesis of the anti-phospholipid syndrome. Among them, interference of anti-phospholipid antibodies with cell acting in the coagulation cascade likely plays a major role. Anti-phospholipid antibodies have been shown to react with endothelial cells mainly by reacting with beta 2 glycoprotein I expressed on the cell membrane surface. Beta 2 glycoprotein I can adhere to endothelial cell surface through the Annexin II receptor and through negatively charged structures (heparin-like molecules) that are bound by the phospholipid-binding site of the molecule. The autoantibody binding involves a yet unknown receptor that activates a signalling pathway able to translocate NFkappaB from the cytoplasm to the nucleus and to activate genes for adhesion molecule, pro-inflammatory cytokine and Tissue Factor up-regulation. The ultimate effect is the induction of a pro-inflammatory and a pro-coagulant endothelial phenotype that has been reproduced both in vitro and in vivo experimental models. Additional effects of anti-phospholipid-mediated endothelial cell activation are the interference with the protein C/S system, with the Annexin V binding, the up-regulation of endothelin I synthesis and the induction of apoptosis. Altogether these effects cooperate in switching endothelium from an anti-coagulant to a pro-coagulant surface. PMID- 12638901 TI - Viral origin of antiphospholipid antibodies: endothelial cell activation and thrombus enhancement by CMV peptide-induced APL antibodies. AB - Our observations and those from others, give further support to our hypothesis that "autoimmune aPL" may be generated by immunization with products from bacteria or viruses after incidental exposure or infection. We also were able to generate APS-like syndrome in a strain of mice susceptible to autoimmunity, indicating that other factors such as genetics are likely to be involved in the development of APS. Furthermore, not all aPL antibodies generated by immunization with bacterial or viral products are pathogenic. Based on the clinical experience and on the numerous reports indicating presence of aPL in a large number of infectious diseases, it may be expected that not all aPL antibodies produced during infection will be pathogenic. We hypothesize that a limited number aPL antibodies induced by certain viral/bacterial products would be pathogenic in certain groups of predisposed individuals. Identification of these bacterial and/ or viral agents may help to find strategies for the prevention of production of aPL "pathogenic" antibodies. Alternatively, free peptides may be used to induce tolerance against aPL production. PMID- 12638902 TI - Antibodies to factor XII: a possible predictive marker for recurrent foetal loss. AB - Antibodies to factor XII (FXIIabs) have been demonstrated in some patients with the anti-phospholipid syndrome (APS). The presence of these antibodies were shown to lead to statistically significantly reduced levels of FXII (p = 0.02). In an extension to this study forty female patients with either primary APS (n = 26) or systemic lupus erythematosus (APS positive) (n = 14) were investigated for levels of factor XII, the presence of lupus anticoagulant and antibodies to cardiolipin, beta 2-glycoprotein I and factor XII. Twenty one of the forty patients had a history of foetal loss (> 2, mean = 2.6). Lupus anticoagulant positivity showed a weak association with foetal loss (odds ratio = 1.1). While there was no association between the presence of antibodies to cardiolipin or beta 2 glycoprotein I with foetal loss, antibodies to factor XII showed a strong and statistically significant association (odds ratio = 5.4, p = 0.025). PMID- 12638904 TI - Nodular regenerative hyperplasia (NRH) of the liver--a manifestation of 'organ specific antiphospholipid syndrome'? AB - Nodular regenerative hyperplasia (NRH) of the liver is a local hyperplastic response of hepatocytes, probably due to vascular abnormalities. Since it was shown in a few case reports that NRH may be associated with antiphospholipid antibodies (APA) we wanted to analyze the relevance of APA in patients with this disease. Sera from 13 patients with histologically defined NRH were tested for APA by an in-house ELISA using as antigens cardiolipin (CL), beta2-glycoprotein I (beta2-gp I), phosphatidylserine (PS), and thromboplastin (TP), a mixture of different phospholipids and phospholipid-binding proteins. As controls, sera from patients with serologically and histologically defined autoimmune liver diseases (primary biliary cirrhosis n = 14; autoimmune hepatitis n = 14) without histological evidence for NRH as well as from 14 healthy blood donors were analyzed. 77% of the NRH patients had APA. In 46% they were directed against CL. In contrast, only 14% of the patients with autoimmune liver diseases and 14% of the healthy controls had anti-CL antibodies (p < 0,05). Antibodies to beta2-gp I and TP did not discriminate between NRH and autoimmune liver diseases. Anti-PS antibodies were not observed. These data indicate that determination of anti-CL antibodies in NRH may help to identify a subgroup of patients in whom an 'organ specific antiphosholipid syndrome' of the liver may be involved in the pathogenesis. PMID- 12638903 TI - Inhibition of ras by farnesylthiosalicylate significantly reduces the levels of autoantibodies in two animal models of the antiphospholipid syndrome. AB - BACKGROUND: Stimulation and proliferation of lymphocytes require activation of Ras. S-farnesylthiosalicylic acid (FTS) is a synthetic substance that detaches Ras from the inner cell membrane and induces its rapid degradation. Antiphospholipid antibodies (aPL) are a heterogeneous group of antibodies detected in patients with antiphospholipid syndrome (APS), which is associated with thrombosis, pregnancy losses, and thrombocytopenia. OBJECTIVE: To examine the effect of FTS treatment on aPL levels in a genetic autoimmune model (the MRL/lpr mice) and in an induced model of APS. METHODS: Female Balb/C mice immunized once with beta2-glycoprotein I (beta2-GPI) in complete Freund's adjuvant (CFA) and female MRL/lpr mice were treated intraperitoneally with either FTS (5 mg/Kg/day) or saline 3-5 times a week. aPL and anti-beta2-GPI antibodies were measured by ELISA. RESULTS: FTS treatment 3 times a week resulted in significant decreases of aPL and anti-beta2-GPI antibodies in both animal models. In contrast, more frequent treatment (5 times a week) had no significant effect on autantibody levels in both animal models. We further compared 2 protocols in the induced APS model, one for alternate day treatment and the other for daily treatment on the first 3 days each week, and found a decrease in autoantibody levels only in the alternate day protocol. CONCLUSIONS: Inhibition of Ras activation by FTS is effective in decreasing autoantibody levels in models of APS. The differential modulation of immune function by alternate day compared to daily treatment may provide better understanding of the role of Ras activation in this system. PMID- 12638905 TI - Frequency and specificities of antiphospholipid antibodies (aPL) in volunteer blood donors. AB - In the State of Indiana, blood donors are screened by a written questionnaire prior to donation to identify potential exposures to infectious diseases and to assess medications. The potential donor is not asked about aPL-related events. Animal experiments have shown, however, that passive transfer of aPL can produce aPL-associated pathology in the recipient. We determined the incidence of antiphospholipid antibodies (aPL) in 775 volunteer blood donors in Indiana. Tubing segments containing 2 ml of anticoagulated blood were obtained from donor units. The average donor age was 43 years (range 17-82); 45% were female. Our in house aPL ELISA tested for IgG, IgA and IgM to cardiolipin (aCL), phosphatidylserine (aPS), phosphatidylethanolamine (aPE) and phosphatidylcholine (aPC). The plasmas were tested with and without supplemental phospholipid (PL) binding plasma proteins from adult bovine plasma (ABP). A total of 24 tests were performed for each donor. Normal ranges were determined for 98% of the donors by calculating multiples of the means (MoM) of OD values for each antigen-isotype combination. No decimals were used, all MoMs were rounded up to the next whole number. An additional two MoMs were added to the calculated cutoff values to eliminate borderline positive values. Results revealed that 63 (8.1 %) donors had positive findings for one or more aPL. The relatively high number of aPL-positive individuals reflects the observation that different donors were often in the highest 2% of each antigen-isotype combination tested. The aPL specificities, isotypes, and PL-binding protein dependence are summarized in this report. PMID- 12638906 TI - The combination of different antiphospholipid antibody subgroups in the sera of patients with autoimmune diseases is a strong predictor for thrombosis. A retrospective study from a single center. AB - OBJECTIVE: [corrected] To determine the distribution of different antiphospholipid antibodies (APL-Ab) and their association with thrombosis in patients with autoimmune diseases. METHODS: Clinical data and laboratory features of 30 patients with different autoimmune diseases with positive APL-Ab were retrospectively studied for a period of more than two years. Anti-cardiolipin (aCL), anti-phosphatidylserine (aPS) and anti-beta2-glycoprotein I (abeta2-GPI) antibodies were determined by ELISA. RESULTS: Autoantibodies that target only PS were detected in 53.3% (n = 16) patients, aCL antibodies only were found in one patient (3,3%). In 43.3% (n = 13), aPS were associated with elevated levels of aCL and/or abeta2-GPI antibodies. No thrombotic event occurred in patients with aPS antibodies only compared to 6 patients from the group with different APL-Ab during 808 +/- 92 days of observation. CONCLUSION: The combination of different antiphospholipid antibody subgroups seems to be a predictor for thrombosis. The presence of aPS antibodies without additional aCL or abeta2-GPI is not associated with thrombosis. The measurement of the APL specificities in addition to the aCL antibodies may be important to develop predictive markers for the risk to develop thrombotic events. PMID- 12638908 TI - Mixed feelings on RCVS proposals for lifelong learning and assessment. PMID- 12638907 TI - Exposure of anionic phospholipids serves as anti-inflammatory and immunosuppressive signal--implications for antiphospholipid syndrome and systemic lupus erythematosus. AB - In contrast to necrotic cells, the clearance of apoptotic ones usually is an anti inflammatory process which elicits only a marginal immune response. During apoptosis phosphatidylserine (PS) is exposed on the outer leaflet of the cytoplasmic membrane and serves as target for the PS receptor of phagocytes. The latter is responsible for anti-inflammatory signalling and the induction of TGFbeta. We were interested whether the immunogenicity of apoptotic cells can be increased by masking PS. We observed that treatment of xenogeneic apoptotic cells with annexin V (AxV) significantly increased the humoral immune response against surface epitopes of these cells. Furthermore, AxV-coated irradiated tumour cells were able to elicit a long lasting tumour specific cytotoxic T lymphocyte response. AxV efficiently blocked the uptake of irradiated cells by macrophages but not by dendritic cells. Furthermore, AxV skewed the phagocytosis of irradiated cells towards inflammation. Investigation of patients with autoimmune diseases further supported the role of anionic surface phospholipids for anti inflammatory clearance of apoptotic cells. Impaired clearance and opsonisation with anti-phospholipid-antibodies are discussed to be responsible for the development of systemic lupus erythematosus and anti-phospholipid-syndrome, respectively. Presentation of cryptic epitopes from late apoptotic cells in a proinflammatory context may challenge T cell tolerance. In addition, accumulation of uncleared apoptotic debris in the germinal centres of lymph nodes may result in the survival of autoreactive B cells. PMID- 12638909 TI - Seroepidemiological survey of Bordetella bronchiseptica and canine parainfluenza 2 virus in dogs in Sweden. AB - The prevalence of antibodies against Bordetella bronchiseptica and canine parainfluenza-2 virus (CPiV-2) was investigated in a population of 302 pet dogs in Sweden. Sera were analysed for B bronchiseptica-specific immunoglobulin G by means of an ELISA, and for CPiV-2 specific neutralising antibody by means of a haemagglutination inhibition test. B bronchiseptica had a seroprevalence of 22 per cent and CPiV-2 had a seroprevalence of 28 per cent. The two pathogens did not appear to circulate together. The crowding of dogs together was significantly associated with the seroprevalence of CPiV-2, but not with the seroprevalence of B bronchiseptica. The dogs' ages, gender or their Federation Cynologique Internationale breed group affiliation was not correlated with the seroprevalence of either pathogen. PMID- 12638910 TI - Abattoir-based survey of contagious bovine pleuropneumonia in cattle in Turkey. AB - Blood samples collected from 945 cattle at four local abattoirs in Turkey were examined for contagious bovine pleuropneumonia (CBPP) by the complement fixation test (CFT) and competitive ELISA (cELISA). In addition, the carcases of the animals were examined macroscopically at the abattoirs and 62 lung samples which had lesions suggestive of CBPP were collected for bacteriological culture. To identify suspicious isolates the PCR was used in addition to the routine biochemical tests. By the CFT, two of the 945 serum samples were seropositive, and by the cELISA, four of them were seropositive. In the bacteriological culture of the lungs, growth was observed in 18 (29 per cent) of the samples by the observation of turbidity in the broths. However, when these broths were inoculated into an agar base, growth was observed in only three (4.8 per cent) samples. These isolates were identified as Mycoplasma species on the basis of biochemical tests. In the PCR analysis of DNA extracted from the broths, none of the isolates was identified as Mycoplasma mycoides subspecies mycoides small colony or one of the members of the M mycoides cluster, but amplification was obtained in only eight (44.4 per cent) of 18 samples, using Mycoplasma-genus specific primers. These DNA samples were examined further with primers specific to 16S rRNA and were then sequenced and compared with the databanks; DNA homologies at different levels were observed in five samples, with Mycoplasma alkalescens, Mycoplasma canadense, Mycoplasma bovis and Mycoplasma bovigenitalium. PMID- 12638911 TI - Effects of atropine, scopolamine and xylazine on the placement of an orally administered magnet in cows. AB - This study was carried out to determine whether the administration of atropine, scopolamine or xylazine to cows before the administration of a magnet orally would help to position it in the reticulum. The transit time of the magnet through the oesophagus was also measured. Sixty Swiss Braunvieh cows were examined by radiography and ultrasonography to locate the reticulum. They were then divided into six groups of 10. Before the administration of the magnet, a control group received 4 ml saline solution subcutaneously, one group received 0.10 mg/kg of atropine subcutaneously, a second received 0.05 mg/kg of atropine intravenously, a third received 0.15 mg/kg of scopolamine intravenously, a fourth group received 0.02 mg/kg of xylazine intravenously, and the cows in the fifth group were positioned so that their forelimbs were 30 cm lower than their hindlimbs during the administration of the magnet. The passage of the magnet through the oesophagus was timed with a stopwatch and monitored with a compass. In the control group the magnet passed through in less than 60 seconds, but in four of the cows receiving either atropine or xylazine intravenously, or having their forelimbs positioned lower than their hindlimbs, it took longer than 60 seconds. In the cows receiving atropine subcutaneously or scopolamine intravenously, it took the same time as in the control group. All the cows were radiographed one-and-a-half hours after the administration of the magnet to determine its location. In seven of the 10 cows in the control group, the magnet was located in the reticulum, but in the other three it was in the cranial dorsal blind sac of the rumen. In the other five groups the magnet was located in the reticulum of between four and seven of the 10 cows, but in the cranial dorsal sac of the rumen, the rumen or in other sites in the other cows. PMID- 12638912 TI - Effect of body condition of postweaning 'flushed' sows and weaning-to-mating interval on sow reproductive performance. PMID- 12638913 TI - Molecular epidemiology of peste des petits ruminants viruses from southern India. PMID- 12638914 TI - Grade III mastocytoma in a donkey. PMID- 12638915 TI - Launch of a dog and cat exotic disease surveillance scheme. PMID- 12638916 TI - Farm animal disease surveillance. PMID- 12638917 TI - Framework for veterinary education and training. PMID- 12638918 TI - Medication of horses in training. PMID- 12638919 TI - Vaccination in the control of avian influenza in the EU. PMID- 12638920 TI - Ectoparasite awareness in sheep. PMID- 12638921 TI - Feline cystitis. PMID- 12638922 TI - Designing radiotherapy plans with elastic constraints and interior point methods. AB - A new linear programming model used to aid in the design of radiotherapy plans is introduced. This model incorporates elastic constraints, and when solved with a path following interior point method, produces favorable plans. A sound mathematical analysis shows how to interpret the solution, and hence, the treatment planner receives meaningful knowledge about the radiotherapy plan being developed. Preliminary experiments are conducted. PMID- 12638923 TI - The initial effects of Physician Compensation Programs in Taiwan hospitals: implications for staff model HMOs. AB - This paper examines whether a Physician Compensation Program (PCP), which was based on the responsibility centers system, improved departmental efficiency in a large Taiwan teaching hospital. PCPs in Taiwan may have implications for staff model HMOs. Monthly financial data and related information for 58 departments in the 5 months following the introduction of the program (the PCP period) and the corresponding 5 months before the introduction of the program (the pre-PCP period) were provided by the case hospital. The Data Envelopment Analysis (DEA) model is used to measure the operational efficiency of each department in the case hospital over the two periods. We first use asymptotic DEA-based tests to examine whether differences in efficiency scores between the two periods are significant. Then, a multi-factor tobit model is used to examine factors that might explain the observed differences in efficiency. The data of each month in the PCP period (November 1996-March 1997) and the pre-PCP period (November 1995 March 1996) are used to calculate efficiency scores and control for monthly effects. We find that average efficiency improves after the implementation of the PCP, with or without controlling for other related factors. Physicians' seniority and percentage of physicians' service time in the department are associated with improved efficiency. Finally, departments with higher profits and fewer numbers of employees are associated with higher efficiency. The findings suggest that to achieve an increase in hospital efficiency in Taiwan, responsibility centers should be integrated with formal physician compensation programs. Such results have implications for staff model HMOs in the US and their variants in countries with national health insurance. PMID- 12638924 TI - Relevance of outlier cases in case mix systems and evaluation of trimming methods. AB - OBJECTIVES: To determine the most appropriate outlier trimming method when the main source of information for case mix classification is length of stay (LOS) because cost information is unavailable. METHODS: Discharges (35,262) from two public hospitals were analysed. LOS and cost outliers were calculated using different trimming methods. The agreement between cost and LOS trimming was analysed. RESULTS: The trimming method using the geometric mean with two standard deviations (GM2) showed the highest level of agreement between cost and LOS and revealed the greatest proportion of extreme costs. Nearly 5% of cases were outliers, containing 16% of total LOS. This was the best approximation to 18% of extreme cost because when GM2 was applied to LOS, 88% of outlier cost was revealed. CONCLUSIONS: The methods were analysed because they are the most frequently used but the same methodology could be employed to compare other outlier determination methods. Outliers should be calculated because they ought to be valued differently from inlier cases. PMID- 12638925 TI - Evaluating Medicaid HMOs when encounter data are missing: case of developmentally delayed children. AB - In evaluating Medicaid Health Maintenance Organizations (HMOs), crucial information regarding severity of illness of patients is often missing--in part because encounter data are not available. If we assume that patients are either in the HMO or in fee-for-service (FFS) plans (i.e., no in or out migration); then severity of HMO patients can be deduced from encounters of FFS patients. We applied this approach to effectiveness of HMO services for developmentally delayed children. Data supported the assumption of a closed system. Data also showed that over 12 months, severity of FFS patients declined. Therefore, we inferred that the HMO was attracting sicker patients. The HMO was paid less than FFS plan, despite the fact that it attracted sicker patients. PMID- 12638926 TI - An exploratory study using data envelopment analysis to assess neurotrauma patients in the intensive care unit. AB - Patients with severe head trauma were studied retrospectively to determine if Data Envelopment Analysis (DEA) could successfully model patients early in their stay in an intensive care unit. Variables examined were cerebral perfusion pressure, body temperature, mean arterial pressure, serum osmolarity and pCO2. Unlike regression-based models that focus on mean values for the group, DEA evaluates each patient individually calculating an "efficiency" score based on a patient's ability to maximize output for a given set of physiologic inputs. Patients with high efficiency scores were found to have a better chance of making a full recovery than similarly injured patients that were inefficient. This approach needs further study but may offer physicians the opportunity to improve patient outcome subject to the manipulation of individual variables from the results of a DEA model rather than aiming for normal or average physiologic values. PMID- 12638927 TI - Patient flow analysis and referrals--how 1,137 ESRD patients started dialysis during 1998-1999. AB - The study consisted in collecting and analysing data from 15 dialysis centres on referral modalities of 1,137 patients with end-stage renal failure starting renal replacement therapy. All the centres in question had implemented PFA Patient Flow Analysis, a management programme for pre-dialysis care from Baxter designed to help improve the collection of data on dialysis patients, from the first visit to a centre through to the start of renal replacement therapy. The aims of the study were threefold: a) describe patient referral modalities and the eligibility of patients for renal replacement treatment; b) determine how many early referrals (ERs) and late referrals (LRs)* have a permanent PD (Peritoneal Dialysis) or HD (Haemodialysis) access at the first treatment; c) for the main outcomes (permanent access at the first treatment and permanent dialysis treatment), compare the performance of centres that offer enhanced education with those that do not. The main characteristics (sex and age) of ERs (54%) are comparable to those of LRs. However, ERs generally have greater access to PD or the opportunity to choose the dialysis treatment. The vast majority (86%) have permanent access at the first dialysis treatment and a large number (44%) have PD as permanent dialysis treatment. Centres with structured pre-dialysis educational programmes experience a larger number of ERs, therefore ensuring better outcomes. For example, 66.3% of patients at centres with structured pre-dialysis educational programmes (compared to 48.2% at centres without enhanced education) start the dialysis treatment with permanent access; and more patients (40% vs. 22%) receive permanent PD. PMID- 12638929 TI - Evaluation of blood recirculation in catheters for haemodialysis and its relation to blood circuit pressures. AB - The objectives of this study were to evaluate the degree of blood recirculation (REC) in venous catheters for dual lumen haemodialysis, and its relation to the drop in arterial line pressure and return venous pressure during haemodialysis. A total of 25 catheters were evaluated (15 temporary 15 cm catheters, and 10 permanent tunnelled 32 cm catheters). REC was calculated using a blood temperature monitor. With blood flow stabilised at 250 ml/min,the session was initiated and the following was determined: real blood flow, drop in arterial line pressure (pre-pump pressure), positive pressure and blood recirculation. Mean blood recirculation in the total of catheters was 6.4 +/- 3%, mean real blood flow was 230 +/- 9 ml/min. Mean blood pressure was -146 +/- 52 mmHG. Mean positive pressure was 127 +/- 47 mmHG. A strong correlation was found between real flow and arterial pressure (r = 0.79, p < 0.01), and between arterial pressure and blood recirculation (r = -0.57, p < 0.01). Comparing the variables studied, a lower degree of REC (5.2+/-2 vs. 8.5+/-3, p < 0.01), a higher real blood flow (234 +/- 7 vs. 223 +/- 7, p < 0.01) and a lower drop in arterial pressure (-127 +/- 49 vs. -180 +/- 40, p < 0.05) were found in temporary catheters than in permanent catheters. No differences were observed in positive pressure. As a conclusion, these results suggest that blood recirculation is lower in temporary catheters than in permanent tunnelled catheters. This is related to a lower drop in pre-pump blood pressure and a higher real blood flow. However, positive pressure does not seem to influence the degree of blood recirculation in venous dual lumen catheters for haemodialysis. PMID- 12638928 TI - Belgian peer review experience on the Achille's Heel in haemodialysis care: vascular access. AB - AIMS: In order to improve the supervision and to evaluate the quality of care in dialysis units, a national project was promoted as a Peer Review. It consisted of systematic, continuous and critical evaluation of the care and the application of international guidelines and compared the reality of care with standards. METHOD: The first chart consisted of the evaluation of infectious episodes of vascular access. This point is particularly relevant since infection represents the second cause of mortality in haemodialysis. A questionnaire concerning each patient was designed. Questions concerned the description of vascular access and the related infectious events. Each questionnaire included 21 items. The project involved 29 dialysis centres, 1,644 patients and 1,775 vascular accesses. The database included 90,525 data. RESULTS: Among the 29 centres, the native arteriovenous fistula (AVF) is the first choice (67.5%) in vascular access, but the proportion of AVF decreases with age contrary to the catheter, which is more frequently chosen, in older patients. Independent of age, 20% of hospitalisations are among patients with catheters and only 7% among patients with AVF. The RR (relative risk) of being hospitalised (any complication of vascular access) is 1.68 for patients with catheters compared to patients with AVF. The rate of infections does not increase with age but is higher for patients with catheters (RR = 2.26). The number of infections appears to be dependent on the staphylococcus aureus carriage in the year. CONCLUSIONS: This first step allows each centre to compare itself to others in an anonymous way. This approach should lead to specific recommendations to improve the quality of care in dialysis units. PMID- 12638930 TI - Causes of late leaks in peritoneal dialysis patients. AB - BACKGROUND/AIMS: Late dialysis leakage is an important complication of peritoneal dialysis. The aim of our study was to investigate the causes of late leaks in patients on peritoneal dialysis. METHODS: 25 patients (19 F: 6 M; mean age: 56 +/ 14) with and 25 patients (10 F : 15 M; mean age: 57 +/- 12) without dialysis leakage on maintenance peritoneal dialysis treatment were included in the study. Data on demographic characteristics including age, sex, body mass index, body surface area, primary renal disease and peritoneal dialysis characteristics including implantation technique of the catheter, site of the catheter, time from catheter implantation to initiation of peritoneal dialysis, daily total filling volumes, use of hypertonic solutions, membrane transport properties and total and peritoneal creatinine clearances were retrospectively collected for both groups. RESULTS: The number of female vs. male patients (76% vs. 40%, p < 0.05) and patients on CAPD vs. APD (68% vs. 56%, p < 0.05) were significantly higher in the group with dialysis leakage. Patients with leakage also had lower average transport properties (p < 0.05) and lower total creatinine clearances (p < 0.05). Other demographic and peritoneal dialysis characteristics were similar in both groups. CONCLUSIONS: Female gender and CAPD as opposed to male gender and APD are the risk factors involved in the development of late dialysis leaks. Peritoneal membrane transport characteristics as well as total creatinine clearance also seems to influence the development of complications. PMID- 12638932 TI - Patient education in renal transplantation. AB - Renal transplantation is the surgical implantation of a human kidney from a compatible donor into a recipient. Renal transplantation is a means of restoring renal function to normal in most patients, thus allowing a return to a healthy lifestyle. Patients and their families often ask nurses for health information. Professional nurses must ensure that patients and families understand the transplantation and therapeutic regimen and provide health education. It should be remembered that certain physiological, psychosocial and pathological conditions might inhibit the education process. Before renal transplantation the patient's understanding of the procedure and follow-up regimen and also the patient's ability to cope with a complex medication regimen are assessed. The nurse and the patient and family should work together to set realistic, achievable goals, the aims of which are mutually agreed. The agreed goals of health education should be documented in a care plan, which will also provide reinforcement for both the nurse and patient. PMID- 12638931 TI - Continuing CAPD after herniotomy. AB - There is still controversy as to whether PD-treatment can be safely continued after herniotomy (HT). Many nephrologists withhold PD-treatment for several weeks after HT in fear of dialysate leakage and/or hernia recurrence. We report on 12 patients (2 women, 10 men) in whom HT was performed either for umbilical (n = 6), inguinal (n = 6) or open processus vaginalis (n = 3). Surgery was performed according to the Lichtenstein method with insertion of a Marlex-mesh and ligation of the hernia sac. In all patients PD treatment was paused for the day of surgery and 1 to 3 days postoperatively, depending on RRF. Low volume (1.0 to 1.5 l) and high frequency exchanges (6 exchanges per day) were started for several days with a gradual reinstitution of the former PD-regimen within the next 2 to 4 weeks. All patients did well rapidly with no uraemia-or dialysis-related complications. No leakage and no hernia recurrence could be observed 3 months thereafter. None of the patients had to be haemodialysed intercurrently. In conclusion, continuing a modified regimen of PD-treatment after HT seems to be safe and comfortable for the patient. PMID- 12638933 TI - In our haemodialysis unit when using the Critline haemoscan, there are problems using this device when administrating transfusions. Any ideas on how to solve this? PMID- 12638934 TI - Has anyone got any information on using CCPD + one HD a week for some one who cant stand HD! How do we calculate their Kt/V? any ideas please? PMID- 12638935 TI - Iron management in renal failure patients--how do we achieve the best results? AB - The majority of patients with renal insufficiency suffer from a normochromic, normocytic anaemia. This renal anaemia affects the quality of life of these patients and reduces their chance of survival. Despite the availability of recombinant human erythropoetin for the treatment of renal anaemia, many patients do not achieve the target haemoglobin concentration of more than 11 g/dl. A major factor contributing to suboptimal correction of renal anaemia and reduced responsiveness to epoetin is a lack of iron availability to the stimulated erythropoesis.Thus, forthe majority of patients with renal insufficiency intravenous iron therapy will be the treatment of choice to replete and maintain adequate iron stores. PMID- 12638936 TI - Do demographic variables affect the timing of referral to the nephrologist? AB - The consequences of late referrals for nephrological care include: increased morbidity, poorer quality of life on dialysis and probably increased mortality. Few studies look at the socio-demographic factors which influence referral to the nephrologist. There is good evidence from studies in other areas of health care that socio-demographic and economic factors influence access to health care. It is important that the nephrology community understand whom the individuals are who likely to be referred late so that we can address any inequality in access to services. We studied all of the patients who started renal replacement therapy in our unit over a five-year period, 1st January 1996 to 31st December 2000 (n = 494). We collected data on gender, age at referral, ethnicity, the date that the individual started dialysis as well as the date they were first seen by a nephrologist. We analysed the data to see if age, gender or ethnicity was associated with timing of referral. If an individual had seen a nephrologist more than three months prior to starting dialysis, they were termed 'early referred', if not they were termed 'late referred' Since this was a sociologically driven research project, we set statistical significance at the 10% (0.1) level. Our data showed that gender did not affect the timing of referral (p = ns), ethnicity affected referral in so much as whites were more likely to be referred late than blacks (p = 0.08) but no more so than non-whites (p = ns). People under the age of 30 were statistically more likely to be referred late than people over the age of 30 years (p = 0.027) as were people under the age of 40 (p = 0.047). We interpret these finding as demonstrating that health care professionals are referring older people and people from the black community in good time and that, in contrast to other studies of inequalities in health, these findings demonstrate that the elderly and ethnic minorities are not being disadvantaged. PMID- 12638937 TI - Emergency management of inherited metabolic diseases. AB - Inherited metabolic diseases with acute severe manifestations can be divided into five categories: (1) disorders of the intoxication type, (2) disorders with reduced fasting tolerance, (3) disorders with disturbed energy metabolism, (4) disorders of neurotransmission and (5) disorders in which no specific emergency treatment is available. Diagnostic emergency laboratory evaluation should cover all differential diagnoses that are therapeutically relevant and should always include ammonia, glucose, lactate and acid-base status as well as testing the urine for ketones. These are indispensable for planning and conducting the first steps of metabolic emergency treatment and should be available within 30 min. According to the clinical situation and biochemical derangement, special metabolic investigations must be initiated in parallel. These include acylcarnitine profiling with tandem mass spectrometry (in plasma or dried blood spots) and analysis of amino acids in plasma and of organic acids in urine. The results of all laboratory investigations relevant to the diagnosis of metabolic disorders for which specific emergency therapy exists should be available within 24 h. There is general agreement with regard to some therapeutic strategies that are clearly explained by pathophysiology: in disorders with endogenous intoxication, anabolism must be promoted and specific detoxification measures initiated. In disorders with reduced fasting tolerance, administration of glucose at the rate of hepatic glucose production forms the basis of treatment. Correction of acidosis is a major goal in disorders with disturbed mitochondrial energy metabolism, while glucose supply may have to be limited. Many current therapeutic strategies are based on case reports and personal experiences at different metabolic centres. The aim of devising the 'best' management is often hampered by the lack of objective evidence of efficacy. PMID- 12638938 TI - Glucose-free medium exacerbates microvesicular steatosis in cultured skin fibroblasts of genetic defects of fatty acid oxidation. A novel screening test. AB - Skin fibroblasts from patients with various fatty acid oxidation defects (FAOD) and four normal controls were subcultured in standard glucose-containing medium or in glucose-free medium simulating fasting. The FAOD fibroblasts developed microvesicular steatosis, which was greatly exacerbated in glucose-free medium. 'Rescue treatment' with glucose-containing medium was performed in the short chain L-3-hydroxyacyl-CoA dehydrogenase-deficient (SCHADD) fibroblasts and resulted in a partial resolution of the steatosis and improved cellular viability. Transmission electron microscopy of autopsy specimens from the SCHADD patient demonstrated that most renal interstitial fibroblasts and approximately 50% of fibroblasts in the heart had microvesicular steatosis. The demonstration of microvesicular steatosis in parenchymal and/or cultured skin fibroblasts may provide important and cost-effective screening tools for the detection of genetic defects of fatty acid oxidation. PMID- 12638939 TI - Identification and characterization of novel mutations of the aspartoacylase gene in non-Jewish patients with Canavan disease. AB - Canavan disease, an inherited leukodystrophy, is caused by mutations in the aspartoacylase (ASPA) gene. It is most common among children of Ashkenazi Jewish descent but has been diagnosed in many diverse ethnic groups. Two mutations comprise the majority of mutant alleles in Jewish patients, while mutations in the ASPA gene among non-Jewish patients are different and more diverse. In the present study, the ASPA gene was analysed in 22 unrelated non-Jewish patients with Canavan disease, and 24 different mutations were found. Of these, 14 are novel, including five missense mutations (E24G, D68A, D249V, C152W, H244R), two nonsense mutations (Q184X, E214X), three deletions (923delT, 33del13, 244delA), one insertion mutation (698insC), two sequence variations in one allele ([10T>G; 11insG]), an elimination of the stop codon (941A>G, TAG-->TGG, X314W), and one splice acceptor site mutation (IVS1 - 2A>T). The E24G mutation resulted in substitution of an invariable amino acid residue (Glu) in the first esterase catalytic domain consensus sequence. The IVS1 - 2A>T mutation caused the retention of 40 nucleotides of intron 1 upstream of exon 2. The results of transient expression of the mutant ASPA cDNA containing these mutations in COS-7 cells and assays for ASPA activity of patient fibroblasts indicated that these mutations were responsible for the enzyme deficiency. In addition, patients with the novel D249V mutation manifested clinically at birth and died early. Also, patients with certain other novel mutations, including C152W, E214X, X314W, and frame shift mutations in both alleles, developed clinical manifestations at an earlier age than in classical Canavan disease. PMID- 12638940 TI - Simple method for detection of mutations causing hereditary fructose intolerance. AB - Aldolase B is critical for sugar metabolism, and a catalytic deficiency due to mutations in its gene may result in hereditary fructose intolerance (HFI) syndrome, with hypoglycaemia and severe abdominal symptoms. This report describes two cases of HFI, which were identified by intravenous fructose tolerance test and a new RFLP (restriction fragment length polymorphism) test that detects the two most common mutations, A149P and A174D. The method includes PCR of a 224-base pair segment of exon 5, a subsequent 3 h incubation with Cac8I and agarose electrophoresis, which reveals either or both of the mutations in one single reaction. The method might be useful for screening of these mutations, which may account for more than 70% of the mutations causing HFI. PMID- 12638941 TI - Glutathione synthetase deficiency: is gamma-glutamylcysteine accumulation a way to cope with oxidative stress in cells with insufficient levels of glutathione? AB - Glutathione (GSH) plays a major role in the cellular defence against oxidative stress and other vital cellular functions. It therefore seems inevitable that patients with severe depletion of GSH will not survive. However, at least some with glutathione synthetase (GS) deficiency do. This study was done to determine whether these patients have a mechanism to compensate for their GSH deficiency. Cell-free extracts of cultured fibroblasts from 9 patients with GS deficiency and 9 control subjects were analysed by HPLC for low-molecular-weight thiol compounds. The patients' cells contained 7.4 nmol of GSH per mg of protein (median; range 2.8-25.2) compared to 33.0 nmol in control fibroblasts (range 26.7 51.4) (p < 0.01). On the other hand, the patients' cells accumulated 18.1 nmol of gamma-glutamylcysteine (gamma-GC) per mg of protein (median; range 6.9-71.7), whereas the control cells contained 0.1 nmol (range 0.05-0.16) (p < 0.01). The cysteine concentrations in the patients' cells were 20.7 nmol/mg protein (median; range 9.4-52.9) compared to 8.9 nmol in control cells (range 3.0-12.4) (p < 0.01). Cultured fibroblasts from patients with GS deficiency have low levels of GSH, but instead accumulate gamma-GC. We suggest that gamma-GC, which contains both reactive groups of GSH (i.e. the sulphydryl and gamma-glutamyl groups), can compensate for GSH in the cellular defence against oxidative stress. Thus, gamma GC may alleviate, but only partly prevent, serious consequences of insufficient GSH levels in affected patients. Since the sum of the levels of GSH and gamma-GC in GS-deficient cells (median 31.5 nmol/mg protein, range 16.2-79.0) was similar to the level of GSH alone in control cells (33.0 nmol/mg protein, range 26.7 51.4), we propose that the cultured fibroblasts may have a mechanism to regulate in a coordinated way the levels of GSH and gamma-GC; for instance, by both compounds acting as feedback inhibitors of gamma-GC synthetase. PMID- 12638942 TI - Mutation analysis of the acid ceramidase gene in Japanese patients with Farber disease. AB - Farber disease is a rare lysosomal storage disease, characterized by the accumulation of ceramide in tissues due to acid ceramidase deficiency. Here we report the identification of three novel mutations in the acid ceramidase gene from two Japanese patients. Patient 1 showed joint problems at around 10 months of age and the patient is now emaciated, with multiple nodules and mild neurological problems at 10 years of age. Patient 2 had consanguineous parents and showed joint contractures at around 8 months of age. He showed neurological symptoms around 2 years of age and died at 6 years owing to respiratory failure. The diagnosis was made clinically and was confirmed by enzymatic assay of acid ceramidase. Molecular analysis of cultured skin fibroblasts showed normal mRNA levels expressed in both patients. By direct sequencing of cDNA, missense mutations of V97E in exon 4 and G235R in exon 9 were detected in patient 1 and 96delV in exon 4 was homozygously identified in patient 2. These mutations were also confirmed in genomic DNA. Expression of mutated acid ceramidase cDNA in COS 1 cells showed acid ceramidase activity decreased to 35%, 2% and 37% of control value, respectively. We also found a new polymorphism V3691 in exon 14 in the allele from the mother of patient 1. To date, 13 mutations, including our newly identified mutations, have been reported. All these mutations were genetically private and genotype-phenotype correlations could not be made. PMID- 12638943 TI - Pamidronate treatment of severe osteogenesis imperfecta in a newborn infant. PMID- 12638944 TI - Contributions to the MODY5 phenotype. PMID- 12638945 TI - Newborn screening compared to clinical identification of biochemical genetic disorders. AB - A group of 28 patients with inherited metabolic disease (homocystinuria galactosaemia, maple syrup urine disease and biotinidase deficiency) diagnosed by screening were compared with a group of 17 similar patients identified clinically. The rate of hospitalization was similar for the two groups. The patients diagnosed clinically showed a higher incidence of mental retardation and their parents experienced greater stress and found greater difficulty in meeting their child's needs. PMID- 12638946 TI - G317D mutation in the tissue-nonspecific alkaline phosphatase gene associated with childhood hypophosphatasia in a German family. PMID- 12638947 TI - Deoxyuridine accumulation in urine in thymidine phosphorylase deficiency (MNGIE). PMID- 12638948 TI - A chemist's approach to biochemical complexity. Interview by Susan Aldridge. PMID- 12638949 TI - Renaissance of immobilized catalysts. New types of polymer-supported catalysts, 'microencapsulated catalysts', which enable environmentally benign and powerful high-throughput organic synthesis. AB - Immobilized catalysts have been reinvestigated from two aspects; as keys to environmentally benign chemical processes and high-throughput organic synthesis for combinatorial chemistry. While most known polymer-supported catalysts are less active than the corresponding original catalysts, new types of polymer supported catalysts, microencapsulated catalysts, have been developed. The catalysts were immobilized on to polymers using physical envelopment by polymer backbones and interaction between pi electrons of benzene rings of the polystyrenes used as polymer backbones and vacant orbitals of the catalysts. Microencapsulated Sc, Os, Pd and Ru catalysts have been successfully prepared and high activities have been attained. In all cases, no leaching of the catalysts occurred, and the immobilized catalysts were recovered quantitatively by simple filtration and reused without loss of activity. It is noted that this method enables direct immobilization of metals onto polymers, and that normally unstable species such as Pd(0)(PPh3) can be kept stable by this immobilization technique. It is expected that other metal catalysts can be immobilized using this microencapsulation technique. PMID- 12638950 TI - Synthesis and biological evaluation of spongistatin/altohyrtin analogues: E-ring dehydration and C46 side-chain truncation. AB - Simplified analogues of the potent antimitotic marine macrolide spongistatin 1/altohyrtin A were synthesised and evaluated as growth inhibitory agents against a range of human tumour cell lines, including Taxol-resistant strains, revealing that E-ring dehydration leads to enhanced cytotoxicity at the low picomolar level while truncation of the side-chain at C46 results in a drastic decrease in activity. PMID- 12638951 TI - Redox-robust pentamethylamidoferrocenyl metallodendrimers that cleanly and selectively recognize the H2PO4- anion. AB - The first pentamethylferrocenyl (Fc*) dendrimers are synthesized from DSM polyamine dendrimers (generations 1 to 5) and cleanly and selectively recognize the H2PO4- anion. PMID- 12638952 TI - The Suzuki coupling of aryl chlorides in TBAB-water mixtures. AB - Palladium acetate in a mixture of TBAB and water can be used as an effective catalyst for the Suzuki coupling of deactivated aryl chloride substrates. PMID- 12638953 TI - A 2,3-butanedione protected chiral glycine equivalent--a new building block for the stereoselective synthesis of enantiopure N-protected alpha-amino acids. AB - A new chiral glycine equivalent 7 has been synthesised from glycidol using a chiral memory protocol, and its use in the synthesis of N-Z protected alpha-amino acids was demonstrated in a series of diasteroselective lithium enolate alkylation reactions and subsequent acid hydrolyses. PMID- 12638954 TI - Ammoximation of ketones catalyzed by titanium-containing ethane bridged hybrid mesoporous silsesquioxane. AB - The first example of ammoximation of bulky ketones over highly hydrophobic titanium incorporated ethane bridged hybrid mesoporous silsesquioxane with high selectivity of oximes is presented. PMID- 12638955 TI - Molecular tectonics: infinite cationic double stranded helical coordination networks. AB - Upon crystallisation of two bismonodentate tectons based on two pyridine units, interconnected at the meta position by a tetra- or hexa-ethylene glycol fragment and Ag+ cation, double stranded helical infinite coordination networks were formed and structurally characterised. The cationic double helical architectures obtained may be regarded as analogues of DNA in terms of topology. PMID- 12638956 TI - Systematic manipulation of surface chemical reaction on the nanoscale: a novel approach for constructing three-dimensional nanostructures. AB - Nanoscale patches, created by nanografting a maleimide-terminated thiol into a self-assembled monolayer, were elaborated by sequential chemical reactions. Each stage in the nanofabrication was followed by atomic force microscopy (AFM), providing a controlled approach to the fabrication of novel three-dimensional (3D) surface nanostructures. PMID- 12638957 TI - Liquid clathrate formation in ionic liquid-aromatic mixtures. AB - 1-Alkyl-3-methylimidazolium containing ionic liquids with hexafluorophosphate, bis(trifyl)imide, tetrafluoroborate, and chloride anions form liquid clathrates when mixed with aromatic hydrocarbons; in the system 1,3-dimethylimidazolium hexafluorophosphate-benzene, the aromatic solute could be trapped in the solid state forming a crystalline 2:1 inclusion compound. PMID- 12638958 TI - Structural characterization of the first hydrothermally synthesized plutonium compound, PuO2(IO3)2 x H2O. AB - Single crystals of PuO2(IO3)2 x H2O were synthesized under hydrothermal conditions (180 degrees C) representing the first structurally characterized transuranium iodate. PMID- 12638959 TI - Soluble polymer-supported convergent parallel library synthesis. AB - Soluble polymer-supported convergent synthesis has for the first time been successfully exploited for parallel library synthesis; sub-libraries of tripeptide iodoarenes and arylboronic acids reacted smoothly in a multipolymer PdII-catalyzed Suzuki coupling reaction to generate a library of bisaryl-linked hexapeptides. PMID- 12638960 TI - Fluoride-induced chemiluminescent decomposition of 1,2-dioxetanes bearing a phenyl moiety substituted with a methyl having an electron-withdrawing group. AB - A new type of dioxetane bearing a 3-(cyanomethyl)phenyl (1a), 3 (methoxycarbonylmethyl)phenyl (1b), 3-(benzoylmethyl)phenyl (1c), or 3 [cyano(methoxycarbonyl)methyl]phenyl group (1d) decomposed through an intermediary dioxetane bearing a benzylic carbanion to afford crimson to yellow light on treatment with TBAF in DMSO. PMID- 12638961 TI - Self-organized formation of a hierarchical self-similar structure with calcium carbonate. AB - Self-organized growth of calcite crystals in a supersaturated solution with a gradual increase in the influence of silicate anions resulted in the formation of a hierarchical self-similar morphology consisting of three-pointed stars with sizes ranging from 100 nm to 100 micrometer. PMID- 12638962 TI - A novel and efficient method for the Pd-catalysed oxidative carbonylation of amines to symmetrically and unsymmetrically substituted ureas. AB - A new method for the Pd-catalysed oxidative carbonylation of amines to symmetrically and unsymmetrically substituted ureas with unprecedented catalytic efficiencies for this kind of reaction has been developed. PMID- 12638963 TI - Synthesis, spectral and electrochemical investigations of bichromophoric pentads possessing tetraazaporphyrin and (bipy)2RuII/(phen)2RuII moieties. AB - Four (bipy)2RuII and (phen)2RuII moieties have been attached to the magnesium tepraazaporphyrin periphery by exploiting thioether coordination to obtain pentads 1 and 2 respectively. These molecules exhibit strong inter-chromophore electronic interaction as reflected by the changes in their spectral and oxidation potential shifts. PMID- 12638964 TI - Bond energy M-C/H-C correlations: dual theoretical and experimental approach to the sensitivity of M-C bond strength to substituents. AB - DFT methods are used to quantify the relationship between M-C and H-C bond energies; for MLn = Re(eta5-C5H5)(CO)2H and fluorinated aryl ligands, theoretical and experimental investigations of ortho-fluorine substitution indicate a much larger increase in the M-C than in the H-C bond energy, so stabilising C-H activation products. PMID- 12638966 TI - A new organic superconductor, (DODHT)2BF4 x H2O. AB - In addition to two organic superconductors (DODHT)2X [DODHT = (1,4-dioxane-2,3 diyldithio)dihydrotetrathiafulvalene; X = PF6 and AsF6] previously reported by us, the BF4 salt of DODHT containing one water molecule [(DODHT)2BF4 x H2O] has been found to undergo a superconducting transition at 3.2 K under a hydrostatic pressure of 15.5 kbar. PMID- 12638965 TI - Effect of water on the catalytic behaviour of VPO in the selective oxidation of propane to acrylic acid. AB - The role of water in propane oxidation to acrylic acid is associated to slow structural modifications of VPO enhancing catalytic performances. PMID- 12638967 TI - A novel approach to polymeric hollow nanospheres with stabilized structure. AB - Polymeric hollow nanospheres with stabilized structure have been realized by self assembly of crosslinkable rigid poly(amic acid) ester (PAE) oligomer and coil like poly(4-vinylpyridine) (PVPy) via hydrogen bonding in their common solvent and subsequent photo-crosslinking of the PAE. PMID- 12638968 TI - A novel role of zeolite NaY in the thermal reaction of alkyl aryl selenoxides in its supercages. AB - Thermal reaction of alkyl aryl selenoxides in the presence of water or methanol in the supercage of zeolite NaY gave beta-hydroxy- or beta-methoxyalkyl aryl selenides, respectively, and NaY played a novel role to stabilize reactive ArSeOH and to separate an anion of -OH from a carbonium ion which was simultaneously present with the -OH in a supercage. PMID- 12638969 TI - Designed layer assembly: a three-dimensional framework with 74% extra-framework volume by connection of infinite two-dimensional sheets. AB - A coordination polymer with 74% extra-framework volume is prepared by predictable linking of the honeycomb network to generate a framework-structured solid designed with two distinct connecting ligands. PMID- 12638970 TI - Head-to-tail dimerization of acrylates catalyzed by iridium complexes. AB - Head-to-tail dimerizations of acrylates and vinyl ketone were successfully performed by the use of iridium complexes in good yields. An iridium hydride complex generated in situ from [IrCl(cod)]2 and alcohols in the presence of Na2CO3 and (MeO)3P was found to be an active species promoting the head-to-tail dimerization of acrylates. Thus, butyl acrylate afforded the corresponding head to-tail dimer in 86% yield. PMID- 12638971 TI - Transformation of a C-methylcalix[4]resorcinarene-based host-guest complex from a wave-like to a novel triangular brick-wall architecture. AB - A novel 2D triangular brick-wall framework based on CMCR and bpe with included ruthenocene guest molecules is formed with time by conversion of a 1D wave-like polymer structure with an accompanying bowl-to-boat conformational change of the CMCR molecules. PMID- 12638972 TI - A novel, solventless reductive coupling of carbonyl compounds by alkali metals, catalysed by bromobenzene. AB - Catalysed by bromobenzene, alkali metals including Li, Na, and K can mediate the reductive coupling of carbonyl compounds under mild, solventless conditions to furnish the corresponding pinacol products. PMID- 12638973 TI - Direct analysis of catalysts immobilised in ionic liquids using electrospray ionisation ion trap mass spectrometry. AB - Catalysts immobilised in ionic liquids have been analysed using electrospray ionisation ion trap mass spectrometry, which allows identification of the catalyst without contamination from the ionic liquid; additionally, a new ionic liquid containing the Monsanto catalyst as the anionic component, [Rh(CO)2I2]-, has been characterised using the same method. PMID- 12638974 TI - Mono(sulfido)-bridged mixed-valence nitrosyl complex: protonation and oxidative addition of iodine across the Ir(II)-Ir(0) bond. AB - Treatment of [Cp*IrH(SH)(PMe3)] (Cp* = eta5-C5Me5) with [IrCl2(NO)(PPh3)2] in the presence of triethylamine yielded the sulfido-bridged Ir(II)Ir0 complex [Cp*Ir(PMe3)(mu-S)Ir(NO)(PPh3)], which further reacted with I2 and triflic acid to give the diiodo complex [Cp*Ir(PMe3)(mu-I)(mu-S)IrI(NO)(PPh3)] and the hydrido complex [Cp*Ir(PMe3)(mu-H)(mu-S)Ir(NO)(PPh3)][OSO2CF3], respectively. PMID- 12638975 TI - First example of a tetra-carboxylate bridged dimanganese species. AB - The compound [Mn(tda)(bipy)] (tda = S(CH2COO)2(2-)) features the first structurally characterized tetra-carboxylate dimanganese antiferromagnetic system with 'copper acetate' core; the binuclear units, completed by bipyridine chelates, are doubly chained by the tda anions that adopt an unprecedented mu,mu' eta1,eta1,eta1,eta1 coordination mode. PMID- 12638976 TI - Reduction of carbonyl groups to the corresponding methylenes with Ni-Al alloy in water. AB - The reduction of carbonyl compounds 1a-h using Ni-Al alloy in water under reflux proceeded to give the corresponding methylene compounds 2a-h within 2 h in 89.0 99.8% relative yields. PMID- 12638977 TI - Unusual structure of the dimeric 4-bromocalcimycin-Zn2+ complex. AB - The X-ray structure of [Zn(4-bromocalcimycin)2 x H2O] complex shows two highly different conformations of the ligand in the dimeric association, unusual in this ionophore family. PMID- 12638978 TI - A miniature biofuel cell operating at 0.78 V. AB - We report the highest voltage miniature biofuel cell to date, a membrane-less cell operating at 37 degrees C in pH 5 buffer at 0.78 V. PMID- 12638979 TI - Aqueous organometallic reactions of rhodium porphyrins: equilibrium thermodynamics. AB - The reactivity and equilibrium thermodynamic studies of tetra-p-sulfonatophenyl porphyrin rhodium hydride ([(TSPP)Rh-D]-4) with CO, aldehydes and olefins that produce formyl, alpha-hydroxyalkyl and alkyl complexes have been explored in water and compared with the related reactions in non-aqueous media. PMID- 12638980 TI - Yttrium alkyl complexes with a sterically demanding benzamidinate ligand: synthesis, structure and catalytic ethene polymerisation. AB - The benzamidinate yttrium dialkyl complexes [PhC(NAr)2]Y(CH2SiMe3)2(THF)n (Ar = 2,6-diisopropylphenyl; n = 1, 2) were prepared; when activated with [PhNMe2H][B(C6F5)4], the species with n = 1 polymerises ethene to polyethene with a narrow polydispersity. PMID- 12638981 TI - Novel and efficient chiral sulfideoxathiane ligands for palladium-catalyzed asymmetric allylic alkylation. AB - Easily prepared, chiral sulfideoxathiane ligands are described which give excellent enantioselectivity (up to 99% ee) in the Pd-catalyzed allylic alkylation of 1,3-diphenyl-2-propenyl acetate with a range of alkyl malonate nucleophiles. PMID- 12638982 TI - Formal radical cyclization onto benzene rings--a general method proceeding via cross-conjugated dienones. AB - Cross-conjugated dienones of type 5 (X = I), which are readily available from phenols, undergo radical cyclization (5 --> 6 --> 7), and the products are easily aromatized (7 --> 8), giving substances that are formally derived by radical cyclization onto a benzene ring (cf. eqn. (1)). PMID- 12638984 TI - Beta zeolite supported on silicon carbide for Friedel-Crafts fixed-bed reactions. AB - Beta zeolite supported on silicon carbide, with high thermal conductivity and high mechanical strength, was successfully used as an active and stable catalyst for Friedel-Crafts reactions in a fixed bed configuration. PMID- 12638983 TI - The first NH aldimine organometallic compound. Isolation and crystal structure. AB - The first NH aldimine organometallic derivative is unexpectedly formed by the cleavage of the nitrogen-carbon bond of the amino acid fragment of the Schiff base 2,4,6-Me3C6H2CH=NCH(CH2Ph)COOEt when the imine is treated with palladium acetate. PMID- 12638985 TI - A novel cationic heteropolyoxovanadium(IV) cluster functionalized with organic ligands: synthesis and characterization of the fully reduced species [Mn(II)V(IV)6O6[(OCH2CH2)2N(CH2CH2OH)]6]Cl2. AB - The solvothermal reaction of (HOCH2CH2)3N with [(n-C4H9)4N]3[H3V10O28] and MnCl2 x 4H2O in CH3CN and CH3OH yields a novel cationic heteropolyoxovanadium(IV) cluster, [Mn(II)V(IV)6O6[(OCH2CH2)2N(CH2CH2OH)]6]2+, containing a fully reduced new cyclic [MnV6N6O18] core with the Anderson structure. PMID- 12638986 TI - Palladium(0) catalyzed regioselective carbonyl propargylation across tetragonal tin(II) oxide via redox transmetallation. AB - Facile homopropargylation of aldehydes by propargyl bromides over tetragonal tin(II)oxide and catalytic palladium(0) occurs which is proposed to involve the prior formation of dinuclear allenylpalladium followed by redox transmetallation to beta-SnO. PMID- 12638987 TI - A convenient stereoselective synthesis of trifluoromethyl-substituted polyfunctionalized cyclopropane: synthesis of (+/-)-trans-trifluoronorcoronamic acid. AB - Trifluoromethylated polyfunctionalized cyclopropanes were obtained in high stereoselectivity by reacting 2-bromo-3,3,3-trifluoropropene (BrTFP) with active methylenes. This novel method was further applied to the synthesis of (+/-)-trans trifluoronorcoronamic acid. PMID- 12638989 TI - Purification of dodecanethiol derivatised gold nanoparticles. AB - Monolayer protected gold nanoparticles synthesised by a two-phase method contain a significant amount of tetraoctylammonium bromide (TOABr), the quaternary ammonium salt used as the phase transfer reagent, as a persistently retained impurity. A simple purification protocol is described. PMID- 12638988 TI - Controlled synthesis of homopolymers and block copolymers based on 2 (acetoacetoxy)ethyl methacrylate via RAFT radical polymerisation. AB - Reversible addition-fragmentation chain transfer (RAFT) radical polymerisation enables to synthesise well-defined homopolymers and block copolymers based on 2 (acetoacetoxy)ethyl methacrylate, which are capable to strongly coordinate to metals and metal ions. PMID- 12638990 TI - Stoichiometric synthesis of a pure ferrite from a tailored layered double hydroxide (hydrotalcite-like) precursor. AB - Calcination of a layered double hydroxide precursor containing MgII, FeII and FeIII cations with an Mg2+:(FeII + FeIII) ratio of 0.5 affords a pure ferrite spinel, MgFe2O4, which has a higher saturation magnetization than samples of the same material produced by conventional ceramic routes. PMID- 12638991 TI - An evaluation of college online alcohol-policy information. AB - Excessive and underage drinking by US college and university students continues to be a significant problem. Curtailing the misuse of alcohol on college campuses is an important goal of college and university administrators because of the many negative consequences resulting from alcohol misuse. As part of their prevention programs, US colleges and universities are required by law to make information about their alcohol policies available to students. Often the source of this information is the school's Web site. The authors evaluated the alcohol-policy information that is available on the Web sites of the 52 top national universities listed in the 2002 rankings of US News and World Report. In general, they found that the information was difficult to find, was located in many areas of the Web site, and did not provide complete information about the school's alcohol policy. PMID- 12638992 TI - The association of campus resources for gay, lesbian, and bisexual students with college students' condom use. AB - The author and colleagues examined campus resources for gay, lesbian, and bisexual (GLB) students at 119 US colleges to determine how the presence of such resources affected sexually active students' condom use. A random sample of unmarried, sexually active college students surveyed in 1999 completed questionnaires. Condom use was compared across schools with different levels of GLB resources. Condom use rates, analyzed by sex and sex-partner groups, ranged from 32% to 52%. With additional college variables controlled for, GLB resources were positively associated with the proportion of sexually active students who used condoms (p < .001). This association existed primarily for students with only opposite-sex partners. The author's findings suggest that campus resources for GLB students may be associated with college students' condom use. PMID- 12638993 TI - Prevalence and correlates of alcohol-induced blackouts among college students: results of an e-mail survey. AB - The authors conducted an e-mail survey of 772 college students to learn more about their experiences with blackouts. Approximately half (51%) of those who had ever consumed alcohol reported they had experienced a blackout at some point in their lives, and 40% had experienced 1 in the year before the survey. Among those who drank in the 2 weeks before the survey, nearly 1 in 10 (9.4%) had experienced a blackout during that period. Many later learned that, during the blackout, they had vandalized property, driven an automobile, had sexual intercourse, or engaged in other risky behaviors. Experiencing 3 or more blackouts was associated with a variety of other experiences, including heavier drinking, lower grades, an earlier age of drinking onset, and having others express concerns about their drinking. The female students who reported blackouts during the 2 weeks before the survey drank far less than male students did during this time period, supporting the use of gender-specific definitions of risky drinking. PMID- 12638994 TI - The role of student health centers in community-university collaborations. PMID- 12638995 TI - The agony of ecstasy: responding to growing MDMA use among college students. PMID- 12638996 TI - The failure of higher education to reduce the binge drinking rate. PMID- 12638997 TI - Drinking water contamination in Walkerton, Ontario: positive resolutions from a tragic event. AB - In May 2000, Escherichia coli O157:H7 and Campylobacter jejuni contaminated the drinking water supply in Walkerton, Ontario. Seven people died and over 2,000 were ill as a result. The Ontario Provincial Government set up a judicial Inquiry into the circumstances surrounding the outbreak and also moved quickly to introduce a new Drinking Water Regulation that incorporated some significant requirements for drinking water providers. The Inquiry itself was in three parts: (a) part 1 related to the events that occurred in Walkerton and why the water contamination occurred; (b) part 1A related specifically to the role of the Provincial Government in the event; and (c) part 2 related to the future of drinking water safety in Ontario with potential to influence regulation on a wider basis. A number of other actions were taken after Walkerton. In August 2000, the Ontario Government, through the Regulatory body, the Ontario Ministry of the Environment (MOE) (a) re-issued and revised the Ontario Drinking Water Objectives (ODWO) as the Ontario Drinking Water Standards (ODWS) and (b) introduced new regulations governing drinking water in Ontario--the Ontario Drinking Water Protection Regulation. One of the key features of the Drinking Water Protection Regulation was the requirement to produce an independent Engineers' Report on all water systems. This paper provides a unique perspective on the Walkerton tragedy and its aftermath. The author was active in many aspects of the resulting activity (Chair of the Ontario Water Works Association's (a section of the AWWA) Special Committee involved in Part 2 of the Walkerton Inquiry; author of several of the Engineers' Reports mandated by Regulation; reviewer on behalf of the Regulator of Engineers' Reports submitted by others). The Engineers' Reports were of interest because (1) the drinking water providers (mostly municipalities) were mandated by regulation to complete the Reports by specific dates and are paying for the Reports, (2) the work had to be done by a registered professional engineer who is not an employee of the owner or the operator if a different entity and (3) the engineer had to sign a declaration that the Regulator could rely on the accuracy of the Report. In other words, the Municipality retained the Engineer and paid them to produce the Report--the Engineer essentially carried the liability while the Regulator had the final say in the acceptability of the Report, a sort of eternal triangle of responsibilities. The paper will outline how the drinking water profession in North America worked together to provide the Walkerton Inquiry with the benefit of its experience and knowledge of best practices to the benefit of consumers and the drinking water providers. It will also outline the procedures adopted to produce the independent Engineers' Reports and how the findings are being applied to further improve drinking water safety in Ontario, across Canada and in similar situations around the world. PMID- 12638998 TI - A fatal waterborne disease epidemic in Walkerton, Ontario: comparison with other waterborne outbreaks in the developed world. AB - An estimated 2,300 people became seriously ill and seven died from exposure to microbially contaminated drinking water in the town of Walkerton, Ontario, Canada in May 2000. The severity of this drinking water disaster resulted in the Government of Ontario calling a public inquiry by Mr. Justice Dennis O'Connor to address the cause of the outbreak, the role (if any) of government policies in contributing to this outbreak and, ultimately, the implications of this experience on the safety of drinking water across the Province of Ontario. The circumstances surrounding the Walkerton tragedy are an important reference source for those concerned with providing safe drinking water. Although some circumstances are obviously specific to this epidemic, others are uncomfortably reminiscent of waterborne outbreaks that have occurred elsewhere. These recurring themes suggested the need for attention to broad issues of drinking water security and they present the challenge for how drinking water safety can be managed to prevent such tragedies in the future. PMID- 12638999 TI - The impact of urban discharges on the health-related microbiological quality of untreated surface water: can catchment management solve the problem? AB - Results showed that surface resource water quality in the study catchment was generally of a poor health-related microbiological water quality. This was due to poor quality discharges received from urban developments in the catchment. The quality results implied that several water uses in the catchment would impact negatively on the health of the users. Current developments in setting Resource Quality Objectives to manage the quality of the resources involved do not provide for the various water uses. The particular Catchment Management Agencies would, therefore, not be in a position to judge the health-related status of their surface water resource quality objectively. To alleviate this problem, tentative water-use classifications as well as microbiological criteria are proposed. PMID- 12639001 TI - Phenotypic population characteristics of the enterococci in wastewater and animal faeces: implications for the new European directive on the quality of bathing waters. AB - Faecal pollution of recreational bathing waters may derive from point sources of various wastewaters or from more diffuse sources such as run-off of agricultural wastes. The paper describes the application of population similarity studies to the enterococcal flora of various animal faeces and municipal wastewaters as a means of distinguishing human from animal faecal material. A simplified phenotypic testing technique (PhenePlate, PhP) was used to study the fermentation kinetics of eleven carbohydrates by all bacterial isolates. Enterococcal isolates (1,766) from six sources were investigated. Enterococcal population diversity (measured as Simpson's Diversity Index) in wastewater samples was high (mean D(i) = 0.95) compared with those of non-human faeces. The mean diversity of isolates in seabird faeces was 0.72, in sheep and donkey faeces 0.44, in dog faeces 0.42 and in cattle faeces 0.32. Analysis of population similarity coefficients demonstrated that faeces from sheep and cattle showed the greatest similarity (S(p) = 0.72). Sheep and cattle faeces demonstrated a low similarity to municipal wastewater samples. This would suggest that population similarity studies might be a useful tool for distinguishing the relative contributions of municipal wastewater and agricultural run-off to bathing water pollution. The PhP procedure identified a specific PhP type that appears to have high specificity to non-human faeces. It may, therefore, represent an important tool in source tracking. Additional phenotypic and genotypic analysis of PhP types that demonstrate a high degree of source specificity is required. The benefits and limitations of the use of population similarity studies to distinguish pollution sources are discussed in comparison with other source tracking approaches and the implications of these developments for future European Union legislation on the quality of bathing waters are discussed. PMID- 12639000 TI - Identification and genetic characterisation of Giardia and Cryptosporidium strains in humans and dairy cattle in the Waikato Region of New Zealand. AB - Giardia intestinalis and Cryptosporidium parvum are commonly acquired waterborne parasites but little is known about their transmission cycles with regard to humans and animals. Faecal samples were collected over two time periods within the Waikato region of New Zealand from dairy cattle and humans and all positive isolates were genotyped. Of the 724 faecal specimens examined (354 cows, 304 calves and 66 humans), 16 cows, 32 calves and 5 humans were positive for G. intestinalis. Phylogenetic group 1 was found in 26 G. intestinalis positive dairy cattle with 22 being group 2. One G. intestinalis positive human isolate was group 1 and four were group 2. Of the 724 faecal specimens examined two cows, 33 calves and 66 human specimens were positive for C. parvum. All 35 C. parvum positive dairy cattle exhibited the bovine genotype while the 66 positive humans showed a seasonal shift in the prevalent genotype with the bovine dominating the spring (100%) and the human dominating the late summer period (96%). PMID- 12639002 TI - Elucidation of potential transmission routes of Campylobacter in New Zealand. AB - Campylobacter is the most commonly reported notifiable disease in New Zealand. The cost of Campylobacter infections in the country during 1994 was estimated as dollar 61.7M although the true cost was probably higher. Investigation of the main environmental reservoirs and routes of transmission to humans is necessary to formulate the most appropriate intervention strategies. This project investigated the reservoirs of Campylobacter in a defined geographical area within New Zealand and compared strains isolated from humans and environmental sources within this area as a prelude to investigating the likely transmission routes to humans. Campylobacter jejuni was commonly found in faeces from dairy cows, beef cattle, sheep and ducks, chicken carcasses, sheep offal and surface waters and C. coli was commonly found in sheep faeces. Preliminary analysis of Penner types was suggestive of transmission to humans from dairy and beef cattle and possibly from sheep. PMID- 12639003 TI - The use of chemical and molecular microbial indicators for faecal source identification. AB - Identifying the source of faecal pollution is important to enable appropriate management of faecal pollution of water. We are developing and evaluating a combination of these microbial and chemical indicators better able to identify the source of faecal pollution. These assays make use of a combination of direct PCR, culturing, and colony hybridisation to identify source specific species of Bifidobacterium, Rhodococcus and Bacteroides. In conjunction with assays for (a) fluorescent whitening agents and (b) faecal sterols and stanols, these indicators were able to identify human derived faecal pollution in river water containing inputs from septic tanks, municipal oxidation ponds, farmed animals and feral animals. Differentiating amongst the animal sources was more difficult and will require development of molecular assays for organisms specific to each animal group. PMID- 12639004 TI - Novel bacterial ratio for predicting faecal age. AB - This study presents an extension of ongoing research into the utility of the ratio of colonies isolated on membrane filters during the total coliform test using m-Endo broth media. Investigations into the relative shifts in concentrations of indicator bacterial populations over time, in laboratory-based survival studies conducted with filtered river water, were undertaken. Also, analysis of Kentucky River water quality data collected from the inlet of a local water treatment plant was carried out. Survival studies found that the ratio between the raw concentrations of atypical colonies (AC) and total coliform colonies (TC) was directly related to the amount of time coliform spiked river water had been held in open jars in the laboratory. The AC/TC ratio in the jars would rise from < 1 at the time of coliform spiking to > 200 within 4d. The rise in AC/TC ratio with time in river water was confirmed in the analysis of two years of Kentucky River water quality data where the average AC/TC ratio during months with high river flow (rain) was 3.37 and rose to an average of 27.58 during months with low flow. The average AC/TC ratio during high flow months compared to that of raw human sewage (3.9) and the ratio increased to values associated with animal impacted urban runoff (18.9) during low flow months. PMID- 12639005 TI - Microbiological aspects of an urban river used for unrestricted irrigation in the semi-arid region of north-east Brazil. AB - This study compared the behaviour of pathogenic bacteria (Salmonella and Listeria), faecal indicators (faecal coliforms FC and faecal streptococci FS), somatic coliphages and F-specific bacteriophages in an urban river contaminated with domestic sewage and surface run-off from agricultural and cattle grazing lands. The influence of physical and chemical parameters was also investigated as well as Salmonella and Listeria serotype diversity and drug resistance patterns. Faecal contamination was high (FC = 5 x 10(6) - 4 x 10(3) CFU/100 mL; FS = 4 x 10(5) - 2 x 10(2) CFU/100 mL) but decreased along the river by up to 99.5% following 47% reduction of BOD5 and 91% increase of DO, both associated with the self purification process. Somatic coliphages (6.9 x 10(5) - 1 x 10(3) PFU/100 mL) and F-specific bacteriophages (5.8 x 10(4) - 65 PFU/100 mL) behaved similarly with reductions of 99.85%. Salmonella and Listeria were isolated at all sampling points with highestfrequencies (91-100%) at those with sewage discharge and rural water run-off. The lowest value (35%) occurred at the end of the river where it was (a) wider and shallower, (b) it ran slower and was warmer (29-33 degrees C), (c) the pH was alkaline (8.2-9.9), (d) electrical conductivity (2,200-5,800 microS/cm) and DO (6-13 mg/L) were highest. Pathogen decline did not follow exactly FC and FS reduction patterns, while physical and chemical parameters apparently did not interfere with Salmonella and Listeria survival to the same extent as they did with FC and FS. Somatic coliphages and F-specific bacteriophages did not show more resistance than bacterial indicators. Catchment area contribution seemed to be more significant for pathogens than for indicators and rainy periods increased pathogenic isolation frequency. Five Salmonella serotypes and five serogroups were identified. S. hadar and serogroup E were predominant (50%); both are increasing in Brazil apparently from animal sources. Nearly 25% of Salmonella strains were resistant to at least one of twelve antimicrobials tested. Resistance to tetracycline was common (17%) followed by cefalotine (3%). Five Listeria serogroups were isolated and L. grayi (43%) and L. monocytogenes (9%) were present at all points. Listeria drug resistance rates were 100% for oxaciline followed by clindamicine (97%), tetracycline (34%) and vancomycin (32%). Both pathogenic bacterial strains presented resistance to the same drugs observed in humans and warm blood animals but the high number of sensitive strains and the low numbers of strains resistant to more than one drug was not expected because of the heavy anthropogenic impact in this basin. PMID- 12639006 TI - Scope of potential bacterial agents of diarrhoea and microbial assessment of quality of river water sources in rural Venda communities in South Africa. AB - The microbial quality of several, usually untreated, surface domestic water sources, used by rural communities in the Venda Region of South Africa, was assessed to gauge their fitness for human consumption and to highlight the possible impact of waterborne diseases. The water sources studied were six points on the Levubu River and the rivers Mutale, Ngwedi, Tshinane, Makonde, Mutshindudi and Mudaswali. Total and faecal coliform, heterotrophic bacteria, enterococci and coliphage counts were used as indicators/surrogates to estimate the degree of bacterial and viral contamination respectively by standard methods. The presence of potential bacterial agents of diarrhoea such as Salmonella, Shigella, Campylobacter, Plesiomonas, Aeromonas and Vibrio was also determined. Results showed that the ranges of counts with regard to all the water sources investigated were 2.9 x 10(2) - 6.3 x 10(4) CFU/100 mL for faecal coliforms, 6.0 x 10(2) - 3.7 x 10(4) CFU/100 mL for total coliforms, 1.8 x 10(2) - 1.3 x 10(6) CFU/mL for heterotrophic plate count, 1.0 x 10(1) - 3.7 x 10(4) CFU/100 mL for enterococci and 0-13 PFU/100 mL for coliphages. These values are far higher than the acceptable maximum limits prescribed for South Africa by the Dept of Water & Forestry and the Water Research Commission - 0 CFU/100 mL, 5 CFU/100 mL, 1.0 x 10(2) CFU/mL, 0 CFU/100 mL and 1 PFU/100 mL for faecal coliforms, total coliforms, heterotrophic bacteria, enterococci and coliphages respectively. Salmonella, Shigella, Vibrio cholerae, Campylobacter, Aeromonas and Plesiomonas were isolated from several of the water sources investigated. The use of these water sources for drinking and domestic purposes poses a serious threat to the health and well being of the users and calls for urgent government intervention. PMID- 12639007 TI - Environmental factors influencing the microbiological contamination of commercially harvested shellfish. AB - Filter-feeding bivalve molluscs (such as oysters, clams, mussels and cockles) can concentrate contaminants from the water column. The extent of faecal contamination in shellfish is usually estimated by determining the concentration of faecal coliforms and/or Escherichia coli. Three sample points in each of three geographically separate commercial shellfisheries were selected for analysis for the effect of season, spring/neap and high/low tidal cycles, rainfall and wind direction on the results of routine E. coli monitoring. General linear modelling was used for the analyses. The principle factors affecting the contamination of shellfisheries were season, high/low tidal cycle and rainfall. The effects varied between harvesting areas and between individual sampling points within harvesting areas. Undertaking such analyses for all harvesting areas would contribute to the management of monitoring programmes and assist in the evaluation of potentially contaminating sources, such as sewage discharges. The type of analyses undertaken on E. coli monitoring data would also be pertinent for the analysis of putative viral indicators, such as F+ coliphage, and could be extended to data on bacterial and viral pathogens. PMID- 12639008 TI - The use of IL-6 induction as a human biomarker for inflammatory agents in water. AB - A rapid whole blood culture (WBC) assay system was developed to monitor the inflammatory potential of water samples collected in the Manguang Municipal area, South Africa. Several systems associated with the Modder River catchment area were investigated in this preliminary study to find out whether there were potential problems in this catchment area. Three focus areas were investigated: (a) the impact of the Klein Modder River, which passes through a densely populated residential area, on the inflammatory activity of the Modder River; (b) the efficiency of the treatment plants in eliminating inflammatory activity; and (c) the efficiency of chlorination of treated water in eliminating inflammatory activity that was not eliminated by the treatment system. Water contaminated with inflammatory substances induced the pro-inflammatory hormone interleukin 6 (IL 6). All water samples collected from the Modder River induced IL-6 secretion, and the quantity of IL-6 secreted was dependent on the concentration and origin of the sample. The lowest IL-6 inducing activity for river water was obtained for samples collected near the origin of the river (Rustfontein). Samples at subsequent points downstream from confluence with the Klein Modder River showed a major increase in IL-6 inducing activity. The two treatment plants along the Modder River also received water of different inflammatory activity. Both treatment plants eliminated some of the inflammatory activity during processing, however inflammatory residues remained after processing which might have been potentially harmful for infants and people with suppressed immune systems. Chlorination eliminated most of the inflammatory activity, indicating that chlorination of water was a necessary process in ensuring water of low inflammatory content. PMID- 12639009 TI - Predation of Cryptosporidium oocysts by protozoa and rotifers: implications for water quality and public health. AB - Predation by free-living protozoa and rotifers was investigated as a possible mechanism for the removal of Cryptosporidium parvum oocysts in aquatic ecosystems including wastewater treatment plants. Free-living ciliated protozoa (Stylonychia mytilus, Paramecium caudatum and an unidentified wastewater wetland ciliate), an amoeba (Acanthamoeba culbertsoni) and rotifers, all commonly found in aquatic ecosystems, were exposed to varying doses of C. parvum oocysts. All organisms investigated ingested oocysts. Predation activity and rates of ingestion varied with predator species and prey density. Ciliated protozoa demonstrated greater predation activity than A. culbertsoni or rotifers when exposed to 2 x 10(5) oocyst/mL for up to 3 h. Greatest predation after 1 h exposure was observed in P. caudatum, the largest ciliate, with on average 1.9 oocysts/cell (range 0-9 oocysts/cell). Stylonychia mytilus and the wetland ciliate had a similar mean ingestion of around 0.3 oocysts/cell, with numbers internalised ranging from 0-3 oocysts/cell. Rotifers ingested on average 1.6 oocysts/individual (range 0-7 oocysts/individual) whilst amoebae ingested on average 1.8 oocysts/cell after 2 h exposure (up to 3 oocysts/cell). Grazing activity by P. caudatum was demonstrated at a variety of prey levels ranging from 9 to 9,000 oocysts. Numbers of oocysts internalised by Paramecium frequently exceeded the reported human infective dose of 30 oocysts. In general, numbers of internalised oocysts increased with incubation time of up to 20-30 min although the rate of accumulation was slower at lower dose levels. The significance of predation on the fate of Cryptosporidium oocysts in the environment is discussed. PMID- 12639010 TI - Comparative reduction of Norwalk virus, poliovirus type 1, F+ RNA coliphage MS2 and Escherichia coli in miniature soil columns. AB - Norwalk-like viruses (NLVs) are important agents of waterborne illness and have been linked to several groundwater-related outbreaks. The presence of human enteric viruses, in particular the presence of NLVs, is difficult to detect in the environment. Consequently, surrogate organisms are typically used as indicators of viruses from faecal contamination. Whether traditional bacterial indicators are reliable indicators for viral pathogens remains uncertain. Few studies have directly compared mobility and reduction of bacterial indicators (e.g. coliforms, Escherichia coli) and other surrogate indicators (coliphages) with pathogenic human viruses in soil systems. In this study the mobility and comparative reduction of the prototype NLV, Norwalk Virus (NV), was compared to poliovirus 1 (PV1), a bacterial indicator (E coli, EC) and a viral indicator (coliphage MS2) through miniature soil columns. Replicate, 10 cm deep, miniature columns were prepared using three soils representing a range of soil textures (sand, organic muck, and clay). Columns were initially conditioned, then incubated at 10-14 degrees C, dosed twice weekly for 8 weeks with one column pore volume of virus-seeded groundwater per dose, followed by 8 weeks of dosing with one column pore volume per dose of unseeded, simulated rainwater. Columns were allowed to drain after each dosing until an effluent volume equivalent to an applied dose was collected. Column effluents and doses were assayed for all viruses and EC. Rapid mobility with minimal reduction was observed for all organisms in the sand. Similar reductions were observed in organic muck for most organisms but NV showed a greater reduction. No organisms were shown to pass through the clay columns. Elution of viruses, in particular PV1, from the columns was gradual. After cessation of microbe dosing, E. coli was less detectable than viruses in column effluents and, therefore, unreliable as a virus indicator. PMID- 12639011 TI - Comparative survival of Cryptosporidium, coxsackievirus A9 and Escherichia coli in stream, brackish and sea waters. AB - Discharge of inadequately treated wastewater into streams may result in the dissemination of pathogens and the contamination of surface water sources. Determining the die-off rate of pathogenic microorganisms in stream and sea waters may serve as the basis for evaluating the health risks posed by the presence of pathogens in seawater. This study was conducted to determine the effect of microbial load, temperature, salinity and turbidity on the die-off of oocysts of Cryptosporidium as compared to that of coxsackie A9 virus (Cox A9) and E. coli. The test microorganisms were seeded into stream, outfall or sea waters and incubated at either 30 degrees C (summer) or 15 degrees C (winter). At 30 degrees C, the fastest die-off was observed for Cox A9 where < 5-log was reduced regardless of the water quality. At 1 degrees C Cox A9 persistence was similar to that of Cryptosporidium where no change was detected in the concentration of either throughout the study period. E. coli die-off reached 5 orders of magnitude within 10d then its concentration remained unchanged. The die-off of E. coli was faster than observed for Cox A9 at 15 degrees C regardless of the water quality. No decrease was observed in the viability of Cryptosporidium under all tested conditions throughout the study period indicating the unsuitability of E. coli to serve as an indicator for the presence of parasites and viruses in stream and marine waters. The prolonged persistence of pathogenic microorganisms in marine waters suggested that discharge of contamination into streams may present a serious environmental health risk. PMID- 12639012 TI - Recovery of feline calicivirus infectious particles and genome from water: comparison of two concentration techniques. AB - The aim of this work was to determine the recovery rate of feline calicivirus (FCV-F9) infectious particles and genome from water after a concentration step using either adsorption elution on glass wool or filtration through an electropositive membrane. The results showed that the membrane filtration technique allowed a 75% recovery rate of FCV-F9 infectious particles while the yield was only 5.3% for FCV-F9 genome. Using the glass wool adsorption-elution technique, the recovery rate of FCV-F9 infectious particles was 0.5% while the yield was 102.5% with Poliovirus 1. PMID- 12639014 TI - Detection, enumeration and isolation of strains carrying the stx2 gene from urban sewage. AB - Verotoxigenic Escherichia coli strains have been related with waterborne outbreaks. Besides 0157:H7, several serotypes of E. coli and other enterobacteria have been implicated in outbreaks and reported to carry the shiga toxin genes. Shiga toxins, stx1 and stx2, are important virulence factors of these strains. These genes have been linked to bacteriophages and consequently are susceptible to lateral transmission. To better understand the ecology of these genes a study of the presence of the shiga toxin 2 gene (stx2) among coliform bacteria present in sewage samples was carried out. A procedure based on colony hybridisation was developed for the isolation of enterobacteria carrying this gene. Colony growth on Chromocult agar was transferred to a membrane and hybridised with a gene specific probe. The procedure allowed detection of about one colony carrying the gene among around 1,000 faecal coliform colonies. The numbers of bacteria carrying the gene in sewage were also estimated by PCR indicating that the numbers of bacteria carrying the stx2 gene were about 1/1,000 faecal coliforms. The detected numbers by both methods were similar. Positive colony hybridisation was detected in four sewage origins. Fifty-two colonies showing positive signal were isolated from the Chromocult agar plates, confirmed to be stx2 positive by PCR and phenotypically characterised. Results of the characterisation showed certain diversity among the isolates even in isolates from the same sample. Most of these isolates would not have been isolated with the methods regularly used for the isolation of E. coli 0157:H7 strains. The method will allow study of the numbers and characteristics of bacteria carrying the stx2 gene in different water environments and isolate them in order to determine their role in the spread of the gene. PMID- 12639016 TI - Laser scanning detection of FISH-labelled Escherichia coli from water samples. AB - The development of rapid and accurate methods for the detection and quantification of bacteria without cultivation is of increasing importance for water monitoring. The aim of this study was to develop a solid phase cytometry detection method for DVC-FISH labelled Escherichia coli cells. In order to allow Stomatic detection with ChemScan RDI, the fluorescein-tyramide was combined with an oligonucleotide probe directly labelled with horseradish peroxidase to increase the fluorescence intensity. The method developed was tested for the enumeration of pure cultures, for GAC-filtered and drinking water samples. The method, which appeared to be equivalent to the culture method, was less sensitive than the DVC-FISH method followed by microscopic analysis. Research is underway to further optimise the labelling conditions. PMID- 12639013 TI - PCR as a test for the presence or absence of Legionella in (cooling) water. AB - Legionella pneumophila, a Gram-negative bacterium, is the causative agent of legionellosis. Traditionally, culture methods are normally used to detect Legionella species in different types of water (e.g. surface or tap water, circulating systems, air conditioners and their cooling devices). In this study the PCR conditions to detect Legionella were optimised based on the EnviroAmp Legionella kit (Perkin-Elmer) which is no longer commercially available. The PCR is very sensitive and specific in indicating the presence or absence (no quantification with classical PCR) of Legionella spp in general and more specifically L. pneumophila. To identify L. pneumophila. DNA sequences from the mip (macrophage infectivity potentiator) gene were amplified. The mip gene is conserved and specific for L. pneumophila although mip-like genes are also present in other Legionella spp. The PCR techniques were able to detect small amounts of Legionella in tap water samples. Cooling water, however, often contained PCR-inhibiting substances that could result in false negative PCR results for Legionella. PMID- 12639015 TI - Detection of Naegleria spp. and Naegleria fowleri: a comparison of flagellation tests, ELISA and PCR. AB - To detect Naegleria spp, in particular Naegleria fowleri, the causative agent of human primary amoebic meningoencephalitis, a flagellation test (FT) is routinely used followed by a specific ELISA. A positive FT indicates the presence of Naegleria spp although some false negatives are likely to occur since parameters for enflagellation vary greatly. As negative FTs are not routinely screened any further for the presence of N. fowleri, this could result in an underestimation of the presence of this pathogen. Therefore, amoebae were further analysed using ELISA and standard PCR not only after a positive but also after a negative FT. In this study 39 cultures containing amoebae were tested with FT, ELISA and the two PCR assays with 11 positive for FT. These were submitted to ELISA and four confirmed as N. fowleri. PCR with the common primer-set on these 11 positive FTs revealed all as Naegleria spp. The specific PCR used on these cultures detected four positive for N. fowleri, corresponding totally with the ELISA results. The 28 negative flagellation tests were also submitted to ELISA and PCR. Of these, 11 were identified as Naegleria spp with common PCR and six as N. fowleri as well as with ELISA and the specific PCR. When the detection of Naegleria spp is based on intermediary processes, such as flagellation tests, false negatives are likely to occur leading to severe underestimations. This study has shown that amoebae taken from negative FTs can be identified as Naegleria spp and N. fowleri when using PCR and ELISA. The application of at least one of the specific N. fowleri tests is recommended for routine screening. The heterogeneous distribution of the false negative results between the different power plants suggested the presence of different genotypes. PMID- 12639017 TI - Laboratory testing protocol to identify critical factors in bacterial compliance monitoring. AB - This research focused on providing guidelines for water utilities on the collection and handling of routine bacteriological samples and in developing scientifically-based approaches in selecting the most representative sampling locations. A laboratory-scale pilot distribution system was designed comprising two parallel loops, one using unlined cast-iron pipe and one using PVC pipe. Each loop contained six sampling ports, including (1) a distribution main dead end faucet, (2) one long (5.5 m; 18 feet) and (3) one short (0.3 m; 1 foot) household copper service line with threaded hose-bibb taps, (4) one hose-bibb with welded faucet, (5) one dedicated sampling port (modeled after a manufacturer's specifications) and (6) one laboratory-style (PVC) stop-cock sampling port. Residual chlorine concentrations were maintained at 0, 0.5, 1.5 and 2.0 mg/L stages during the course of the experiment. Bacterial samples were collected from the different sampling ports and assayed by membrane filtration and/or spread plate. Nutrient and R2A agars were used for heterotrophic plate counts (HPC), m Endo agar for total coliform (TC) counts and Chromocult agar for injured bacterial analyses. Several methods of sample collection were tested using various combinations of flushing and tap disinfection, including "first flush" (no flushing, without tap disinfection), flushing only, tap disinfection only (using alcohol or hypochlorite solution) and flushing coupled with tap disinfection. The results indicated that the bacterial counts in samples drawn from dead ends were not significantly different from counts in samples from the other sample port configurations. First flush samples consistently produced the highest bacterial count results. Bacterial counts in samples from the long household copper service line were typically three orders of magnitude higher than in samples from the other sample ports. Thus, there is evidence that long copper household service connections may be unsuitable sample tap configurations for collecting samples intended to represent microbial quality in the distribution system. PMID- 12639018 TI - A chemiluminescent immunofocus assay (CIFA) for non-microscopic enumeration of Cryptosporidium parvum infectivity in cell culture. AB - As Cryptosporidium parvum continues to cause waterborne disease, despite extensive efforts by drinking water suppliers and regulators, it is important to have reliable and convenient methods for detection of this pathogen in wastewater discharges, environmental source waters and finished drinking water supplies. In order to better understand the health risks of this organism, it is necessary that detection methods be able to distinguish between infectious and non infectious Cryptosporidium oocysts in these environmental samples. Cryptosporidium infectivity assay systems based on infections in mice and on in vitro infections in continuous mammalian cell lines are available. Currently, these methods are impractical for routine analysis of water samples because they are tedious, lengthy and costly. These methods rely on careful microscopic examination or further analysis by PCR and then characterisation of the amplified DNA. Practical and affordable non-microscopic methods to determine Cryptosporidium infectivity are much needed for environmental analysis. A cell culture infectivity detection system was developed for infectious Cryptosporidium oocysts that does not rely on microscopic examination of samples to score results, is applicable to a variety of samples and has the potential to be used for routine water monitoring and other environmental or biomedical analysis. Using a chemiluminescent immunoassay, the discrete foci of developmental stages of Cryptosporidium in cell cultures are clearly visible as discrete objects in an image of the entire cell culture layer in a dish or on a slide. These objects are directly countable with the unaided eye and their identity can be further confirmed or verified by microscopic examination. PMID- 12639019 TI - A detection method for Legionella spp in (cooling) water: fluorescent in situ hybridisation (FISH) on whole bacteria. AB - Although traditional culture methods are appropriate for detection of Legionella species, such culture takes several days. Rapid detection (< 24 h) of individual Legionella is possible using fluorescent in situ hybridisation (FISH) on whole bacteria. Water samples were filtered and the concentrated bacteria were immediately detected (without culture) with a fluorescence microscope following appropriate labelling. The detection level was very high and quantification was possible. For the detection of all Legionella spp. the probe LEG705 was used, complementary to a 16S rRNA sequence conserved in all Legionella spp. For specific detection of L. pneumophila the probe LEGPNE1 was used. This probe is designed against a variable domain of the 16S rRNA sequence from L. pneumophila. CY3 and FLUOS labels were tested and CY3 showed clearly detectable bacteria with minimum background staining. This FISH technique is very sensitive, fast, reliable and individual bacteria are easily detected. PMID- 12639020 TI - Inactivation of Salmonella spp. from secondary and tertiary effluents by UV irradiation. AB - The aim of this study was to verify the efficiency of UV irradiation in the inactivation of Salmonella spp. in treated wastewater with different levels of turbidity and exposed to increasing doses of UV irradiation. Experiments were carried out in a batch reactor and in a real scale reactor. Salmonellae obtained from clinical samples were seeded into autoclaved wastewater collected from a wastewater treatment plant (WWTP) comprising an association of a UASB reactor followed by three submerged aerated biofilters (BAF) and one tertiary filter. The results showed that salmonellae were not inactivated in effluents from the UASB reactor indicating that the presence of suspended solids was an important obstacle to UV penetration in bacteria. However, UV irradiation was efficient in inactivating Salmonella of effluents from aerated secondary and tertiary biofilm reactors. PMID- 12639021 TI - Efficiency of autothermal thermophilic aerobic digestion and thermophilic anaerobic digestion of municipal wastewater sludge in removing Salmonella spp. and indicator bacteria. AB - The study is focused on the comparison of autothermal thermophilic aerobic digestion, thermophilic and mesophilic anaerobic digestion, based on long-term monitoring of all processes in full-scale wastewater treatment plants, with an emphasis on the efficiency in destroying pathogens. The hygienisation effect was evaluated as a removal of counts of indicator bacteria, thermotolerant coliforms and enterococci as CFU/g total sludge solids and a frequency of a positive Salmonella spp. detection. Both thermophilic technologies of municipal wastewater sludge stabilisation had the capability of producing sludge A biosolids suitable for agricultural land application when all operational parameters (mainly temperature, mixing and retention time) were stable and maintained at an appropriate level. PMID- 12639022 TI - Removal of microbes from municipal wastewater effluent by rapid sand filtration and subsequent UV irradiation. AB - The elimination of wastewater microbes is often necessary when effluent receiving waters are reused for different purposes e.g. for irrigation or as a raw water source of drinking water. In the present study, rapid sand filtration (SF) combined with the use of polyaluminium chloride coagulation was used as a pre treatment to improve the quality of wastewater effluent before further treatment with UV irradiation. Pilot-scale experiments were run in four treatment plants in Finland. Treatment performance was followed by measuring physical and microbial parameters. Rapid sand filtration reduced suspended solids, turbidity and colour of effluents by about 90%, 70-80% and 20-50% respectively. It also improved the UV transmittance of water by up to 20%. Microbes and phosphorus were reduced by 90-99% and to 0.05 mg/L respectively. UV irradiation further reduced the number of microbes up to 99.9%. The efficiency of UV doses in pilot UV reactors was confirmed with collimated-beam device determinations and with added FRNA phages. More than 99.9% reduction of MS2 was achieved with the dose of 140mWs/cm2 in pilot UV reactors. Rapid sand filtration and the subsequent UV irradiation reduced the number of all the tested microbes to a low level, often below the detection limit. Suspended solids and the water turbidity were reduced to 1-2 mg/L and approximately 1 NTU respectively. PMID- 12639023 TI - Parasite contamination of liquid sludge from urban wastewater treatment plants. AB - This study was performed on sludge samples from 20 wastewater treatment plants located in the north west of France with capacities of 1,000-20,000 inhabitant equivalents. The types of treatment studied were activated sludge low charge with and without denitrification. Respectively, 110 samples of fresh sludge and 84 samples of discharged sludge for spreading were analysed. Globally 78.6% of samples contained helminth eggs belonging to the cestodes (6.1%) and nematodes (93.9%). Most of the nematode eggs detected were viable with 135 positive samples. The distribution, according to genera, indicated a high prevalence of Toxocara eggs (77.4%) followed by Capillaria (13.2%), Trichuris (8.1%) and Ascaris (1.3%). For viable nematode eggs, the concentrations detected ranged from < 1 to 28/4 gDM for fresh sludge and from < 1 to 9.6/4 gDM for discharged sludge. PMID- 12639024 TI - Effects of peracetic acid and monochloramine on the inactivation of Naegleria lovaniensis. AB - Biocidal activities of monochloramine and peracetic acid were studied on cysts of Naegleria lovaniensis. Until recently the most commonly used biocide to disinfect cooling water systems was hypochlorite. Owing to its negative impact on the aquatic environment, ecologically less harmful alternatives have been sought. As the biocidal activity of monochloramine and peracetic acid makes them good candidates for inactivation of pathogenic Naegleria species, these biocides were tested against Naegleria lovaniensis, a relative of the pathogen Naegleria fowleri, as an alternative treatment to hypochlorite. Under laboratory conditions the biocidal activity of hypochlorite was 8- 10x stronger than that of the two investigated substances. Hypochlorite, at a concentration of 0.5 mg/L, killed 100% Naegleria lovaniensis after 1 h exposure (25 degrees C, pH 7.3- 7.4). To achieve similar results with monochloramine and peracetic acid, 3.94 mg/L or 5.33 mg/L had to be used respectively (25 degrees C, pH 8). It was known that the in situ biota of the biofilm, along with any organic material in the water column, had a negative impact on the efficiency of the biocides. There are, however, indications that the relative efficacy of monochloramine and peracetic acid was quite good under such conditions when compared with hypochlorite. PMID- 12639025 TI - Ultrasonic treatment of Cryptosporidium oocysts. AB - The effect of 20 kHz ultrasound on the viability of Cryptosporidium oocysts was investigated. More than 90% of the dispersed Cryptosporidium oocysts could be deactivated in about 1.5 min of continuous sonication. In order to apply this technique to large quantities of contaminated water, quantitative filtration and redispersion of Cryptosporidium oocysts were investigated and found to be easily achievable. The estimated cost of sonication showed that the ultrasound treatment of Cryptosporidium oocysts contaminated water could be a very effective means of "deactivating" Cryptosporidium oocysts. PMID- 12639026 TI - Assessing UV reactor performance for treatment of finished water. AB - Recently, use of low levels of medium- and low-pressure ultraviolet light for successful inactivation of Cryptosporidium parvum oocysts has generated tremendous excitement in the water industry. Accurate delivery of the target dose, lamp performance, sensor stability and impact of water characteristics are some factors that could impact disinfection efficacy, in turn influencing decisions on application of this technology. To this end, American Water Systems, the largest investor owned water utility in the US, has responded to some of these challenges by ascertaining the long-term feasibility of applying UV for treatment of finished water. A 4 x 1 UV reactor with a 12 inch (0.3 m) diameter was installed after granular activated carbon filtration and was operated with a finished water flow rate of 600 gpm (2,700 L/min). Over a 12-month period, various chemical (THM, HAA, UV254, DOC, TOC, metals, nitrate, nitrites) and physical measurements (lamp voltage, current, sensor measurements) were monitored to define their impact (if any) on the operation of the reactor. MS2 bacteriophage challenge studies were conducted with various lamp configurations and lamp age. These inactivation data demonstrated high levels of correlation with controlled bench scale inactivation data. For C. parvum oocysts, bench scale studies were performed with a modified in vitro infectivity assay using HCT-8 cells, an enhanced infectivity protocol and with either immunofluorescence or quantitative PCR based detection. While both assays indicated increasing infections levels of HCT-8 cells with increasing oocyst inocula, UV treatment of oocysts produced markedly different infectivity responses. Based on the data generated in this study, one in vitro infectivity assay was selected to demonstrate > 3 logs inactivation with low UV doses (5 mJ/cm(20-10 mJ/cm2). PMID- 12639027 TI - Inactivation differences of microorganisms by low pressure UV and pulsed xenon lamps. AB - UV disinfection has been applied to water treatment in recent years with low pressure and medium-pressure UV lamps mainly used as the light source. In general, UV disinfection is considered to be inefficient with water of high turbidity because of inhibition of light penetration. Additionally, photoreactivation may be a problem that should be considered in case a disinfected water is discharged to the environment where sunlight causes reactivation. Recently, other types of lamps have been proposed including a flush type lamp (such as a pulsed-xenon lamp) that emits high energy and wide wavelength intermittently. In this study, the difference between inactivation efficiencies by low-pressure UV (LPUV) and pulsed-xenon (PXe) lamps was investigated using two coliphage types and three strains of Escherichia coli. PXe had a suppressive effect on photoreactivation rate of the E. coli strains even though there was no significant effect on inactivation rate and maximum survival ratio after photoreactivation. PXe also had a benefit when applied to high turbidity waters as no tailing phenomena were observed in the low survival ratio area although it was observed in LPUV inactivation. This efficiency difference was considered to be due to the difference in irradiated wavelength of both lamps. PMID- 12639028 TI - Effectiveness of guideline faecal indicator organism values in estimation of exposure risk at recreational coastal sites. AB - Decay rates in coastal water and sediment for the bacterial pathogens Salmonella typhimurium and S. derby were compared in laboratory-based microcosms with results previously obtained for a number of faecal indicators. In general, the decay rates of Salmonella spp. were greater than either enterococci or coliphage in overlying water and sediment. Decay rates of E. coli were similar to Salmonella spp. in overlying water, although greater in sediment. Raised temperature resulted in an increased decay rate for all organisms in the overlying water (and to a lesser extent in the surface sediment layer). It was demonstrated that decay rates for both S. typhimurium and S. derby were greater in overlying water compared with sediment. This suggested that sediments may be acting as a reservoir for pathogenic microorganisms released into the coastal environment during recreational activity and should be considered when estimating environmental exposure. Using measured decay rates and available dose-response data, a quantitative microbial risk assessment (QMRA) utilising Monte Carlo simulation was undertaken to estimate the risk of infection to Salmonella spp. following exposure to recreational coastal water subject to a range of faecal contamination levels. In waters of extremely poor quality, subject to contamination by faecal coliforms (10(6) CFU/100 mL), the maximum probability of infection on the day of an accidental release was above 2.0 x 10(-1) and remained above 1 x 10(-3) for three days following the initial high concentration. PMID- 12639029 TI - Factors influencing faecal contamination in coastal marinas. AB - Recreational pleasure and fishing boats are potential sources of human faecal contamination because the sanitary wastes from boat occupants may be discharged into the surrounding water. The impacts of such faecal contamination from boats is potentially high in marinas where boats are often kept and occupied for varying periods of time. The nature and extent of such faecal contamination from boats in marinas of different design and use is still inadequately understood. In this study the levels of faecal coliform (FC) bacteria were measured in the waters of a confined and an open water marina over a 6 d period encompassing a holiday weekend. Levels of FC rose with increasing occupancy by boats and boaters during the study period and then declined again. FC levels were higher in waters of a confined or basin marina than in waters of an open or unconfined marina. In both confined and open water marinas, FC levels were highest in water samples collected near boats but they also became elevated on occasion in water samples taken a distance of 305 m from boats. Concentrations of FC in marina waters exceeded the standards and guidelines for shell-fishing and primary contact recreation waters. Therefore, greater consideration of human health risks from enteric microbes in marina waters and shellfish is recommended. PMID- 12639030 TI - The role of resuspension in enterococci distribution in water at an urban beach. AB - This study investigated the process and effects of bacterial resuspension on microbiological water quality in a small urban embayment. Water and sediments were sampled for enterococci at a small urban bay, on both irregular and intensive time scales, with a focus on the potential sources of faecal contamination to the system. Distribution of enterococci in sediments was influenced by the location and microbiological quality of major sources of enterococci to the embayment. Stream and storm water contributed the greatest numbers of enterococci and, consequently, high numbers of enterococci were found in both water and sediments surrounding discharge points for these sources. To investigate bacterial resuspension, water samples were collected from within the surf zone (at water depths of 1-1.5 m) as a wave crest passed. Two samples were collected simultaneously at each sampling location at 10 cm above the seabed and 10 cm below the water surface. Samples were analysed for enterococci and data compared with bacterial numbers in adjacent sediments as well as in stream and storm water sources. Vertical distribution data for enterococci in the water column revealed evidence of spatial and temporal variability in bacterial resuspension and the role of wave action was demonstrated. Bacterial resuspension under waves was directly related to weather and wave conditions. The resuspension of enterococci was not detected beyond the surf zone suggesting that wave action was the main cause of resuspension at the study site. PMID- 12639031 TI - The significance of sunlight for the elimination of indicator bacteria in small scale bathing ponds in central Europe. AB - In order to determine the impact of sunlight on the reduction rate of indicator bacteria, bathing waters of different turbidity were artificially contaminated with indicator bacteria and exposed to sunlight. The reduction rates determined for the indicator bacteria showed a significant correlation with radiation intensity and, thus, also the time of day as well as the turbidity of the water. Under conditions of strong radiation intensity and low water turbidity (< 5 FTU), indicator bacteria were eliminated within 1-2 h of contamination with an initial concentration of 10(5) CFU/mL. Moreover, differences between different indicator bacteria were found. Enterococcus faecalis proved to be significantly more resistant to sunlight than the other indicator bacteria tested (Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus). The results of the present study, namely the complete elimination of indicator bacteria within a few hours, showed that sunlight, given an appropriate intensity and good water transparency, was the most important factor in the reduction of hygienically-relevant microorganisms in surface waters and, thus, also in small-scale bathing ponds. PMID- 12639032 TI - Water safety plans for small systems: a model for applying HACCP concepts for cost-effective monitoring in developing countries. AB - The development of water safety plans (WSPs) for small systems should be based on a thorough understanding of the relationships between risk factors and contamination events. This can be achieved through the use of well-designed assessments of water quality that provide better evidence to support the identification of control measures, performance limits, monitoring parameters and verification procedures. Training of community operators is critical to the success of the WSP and the understanding gained from the assessments provides a sound basis for addressing these needs. The WSP approach provides for more effective control of water quality and the use of targeted assessments is cost effective in improving the design of WSPs. PMID- 12639033 TI - Chlorination and safe storage of household drinking water in developing countries to reduce waterborne disease. AB - Simple, effective and affordable methods are needed to treat and safely store non piped, gathered household water. This study evaluated point-of-use chlorination and storage in special plastic containers of gathered household water for improving microbial quality and reducing diarrhoeal illness of consumers living under conditions of poor sanitation and hygiene. Community families were recruited and randomly divided into intervention (household water chlorination and storage in a special container) and control (no intervention) households. Microbes in stored household water were extensively inactivated by 1-5-mg/L doses of hypochlorite. Escherichia coli levels in stored household waters were < 1/100 mL in most intervention households but readily detectable at high levels in control households. Stored water of intervention households was also lower in Clostridium perfringens and heterotrophic plate count bacteria than in control households. The intervention reduced household diarrhoeal illness. In Bolivia, monthly episodes of household diarrhoeal illness were 1.25 and 2.2 in intervention and control families, respectively (P = < 0.002) indicating that 43% of community diarrhoea was preventable by using the intervention. In Bangladesh, mean episodes of child diarrhoea/1,000 d were 19.6 and 24.8 in intervention and control groups respectively (P = < 0.03) indicating that about 24% of observed diarrhoea was preventable by using the intervention. Chlorine disinfection and storage in an appropriate container significantly improved the microbiological quality of non-piped household drinking water and reduced community diarrhoeal disease. Widespread use of this simple treatment and storage system for non-piped domestic water has the potential to dramatically reduce the global burden of waterborne diarrhoeal disease. PMID- 12639034 TI - Water quality problems associated with intermittent water supply. AB - A controlled study was conducted in Lebanon over a period of 12 months to determine bacterial regrowth in a small network supplying the Beirut suburb of Naccache that had a population of about 3,000. The residential area, which is fed by gravity, is supplied twice a week with chlorinated water from two artesian wells of a confined aquifer. A significant correlation was detected between the turbidity and the levels of heterotrophic plate count bacteria (HPC) in the samples from the distribution network as well as from the artesian wells. However, a negative significant correlation was found between the temperature and the HPC count in the samples collected from the source. A statistically significant increase in counts, possibly due to regrowth, was repeatedly established between two sampling points lying on a straight distribution line but 1 km apart. Faecal coliforms were detected in the source water but none in the network except during a pipe breakage incident with confirmed Escherichia coli reaching 40 CFU/100 mL. However, coliforms such as Citrobacter freundii, Enterobacter agglomerans, E. cloacae and E. skazakii were repeatedly isolated from the network, mainly due to inadequate chlorination. A second controlled study was conducted to determine the effect of storage on the microbial quality of household storage tanks (500 L), which were of two main types - galvanized cast iron and black polyethylene. The mean bacterial count increased significantly after 7 d storage in both tank types. A significant difference was found in the mean HPC/mL between the winter and the summer. Highest counts were found April-June although the maximum temperature was reported later in the summer. A positive correlation was established between the HPC/mL and pH, temperature and storage time. PMID- 12639035 TI - Artificial neural network modelling: a summary of successful applications relative to microbial water quality. AB - Artificial neural networks (ANN) are modelling tools that can be of great utility in studies of microbial water quality. The ability of ANNs to work with complex, inter-related multiparameter databases and provide superior predictive power in non-linear relationships suits their application to microbial water quality studies. To date ANNs have been successfully applied (a) for the prediction of peak microbial concentrations, (b) to sort land use associated faecal pollution sources and relative ages of runoff and (c) towards the selection and study of surrogate parameters. Predictions of peak microbial contamination or faecal pollution sources have been greater than 90% accurate. The importance of a subgroup of organisms that are isolated by the total coliform membrane filter test on m-Endo media in defining faecal sources was revealed through parameter selection exercises. The result is the definition of a new bacterial ratio that can be directly related to the age of faecal contamination in animal impacted runoff. PMID- 12639037 TI - Antibiotic selective pressure for the maintenance of antibiotic resistant genes in coliform bacteria isolated from the aquatic environment. AB - Coliform bacteria isolated from the aquatic environment were investigated for antibiotic susceptibility and detailed structures of class 1 integrons. A high proportion of isolates were found to be resistant to sulfamethoxazole, aminoglycosides, and beta-lactams. The 750 (53.6%) isolates were resistant to one or more of the antibiotics tested out of 1,400 coliform bacteria. Based on the MIC of antibiotics and antibiogram, 150 isolates were selected and further studied for class 1 integrons. The intI1 gene was found in 36 (24.0%) of the 150 isolates. Twelve isolates carried the gene cassettes responsible for antibiotic resistance, while no gene cassettes were found in 24 isolates. Seven different genes, dfrA5, dfrA7, dfrA12, dfrA17, aaA2, aaA5, and aad(3'), were detected in gene cassettes. The dfrA and aad genes located on class 1 integrons were responsible for resistance to trimethoprim and aminoglycosides. The remaining 24 coliform bacteria had the incomplete or non-functional class 1 integrons. These results indicated that antibiotic selective pressures may play an important role to maintain gene cassettes of class 1 integrons and in the absence of sustained antibiotic pressures, such as the aquatic environment, coliform bacteria may carry empty or non-functional class 1 integrons. PMID- 12639036 TI - Quantitative risk assessment of Cryptosporidium in surface water treatment. AB - Quantitative microbiological risk assessment requires quantitative data to assess consumer exposure to pathogens and the resulting health risk. The aim of this study was to evaluate data sets on the occurrence of Cryptosporidium oocysts in raw water and on the removal of model organisms (anaerobic spores, bacteriophages) to perform such a risk assessment. A tiered approach was used by first calculating approximate point estimates and when the point estimate was close to the required safety level (10(4) annual risk of infection), fitting the data to probability distributions and Monte Carlo analysis to calculate the distribution of the risk of infection. Sensitivity analysis showed that the variability in the Cryptosporidium data in raw water (largely introduced by the variability of the recovery efficiency of the detection method) determined most of the variance in the risk estimate. PMID- 12639038 TI - Evaluation of Bacillus subtilis and coliphage MS2 as indicators of advanced water treatment efficiency. AB - The assessment of water treatment facilities for their efficiency using alternate indicators is of paramount importance. Current methods for assessing efficiency are limited by the specific characteristics of the microorganisms, such as their different sensitivities to disinfectants. A pilot study was carried out to compare different treatment scenarios for the future upgrade of the Sergio Cuevas Water Treatment plant (the largest in the Caribbean) in San Juan, Puerto Rico. The treatment units under investigation included a coagulation-flocculation sedimentation unit, dual-media filters, micro-filtration units, intermediate ozone injection and contact columns as well as a biological filtration unit. The plant was challenged at different stages of treatment with Bacillus subtilis spores and MS2 coliphages in an attempt to test them as possible alternate indicators of treatment plant performance. These organisms were chosen because of their resistance to disinfection and desiccation, their low analysis costs and ease of detection. The removal of spores and coliphages by each treatment unit tested was calculated by seeding a known concentration (5-7 log10) of spores and coliphages and following the removal or disinfection rates. The seeded indicators were detected using traditional culture techniques. Ballasted clarification was shown to be highly efficient at removing 99.1% (approximately 3 log10) of the spores and 85.1% (approximately 0.86 log10) of MS2. Ozone treatment inactivated 80.37% (approximately 1.4 log10) spores and 99.95% (approximately 3.07 log10) coliphages. The coliphage inactivation rate obtained confirmed data obtained by previous studies indicating that MS2 was less resistant to ozonation than B subtilis spores. The membrane technology had the best efficiency in terms of physical removal of spores achieving over 99.9% (> 3 log10) removal. Coliphage removal mechanisms remain to be determined and will be a future focus of the study. Preliminary results indicate that aerobic spores and coliphages may be useful as indicators to determine the efficiency of different drinking water treatment technologies. PMID- 12639039 TI - Virological control of groundwater quality using biomolecular tests. AB - Deep groundwater, even if generally protected, could be contaminated by surface or rain water infiltration through soil fractures, septic tanks, cesspits, land irrigation, disposal of wastewater and disposal of muds from depuration systems. The sanitary importance of such possible contamination is related to the different uses of the water and it is at the maximum level when it is intended for human use. Routine microbiological analyses do not consider viruses, only bacterial parameters, as contamination indicators. However, it is known that enteric viruses can survive a long time in deep aquifers and that they may not always be associated with bacterial indicators. The virological analysis of waters intended for drinking use is provided only as an occasional control exercised at the discretion of the sanitary authority. Technological difficulties with obtaining data about groundwater viral contamination led to a study to devise rapid and efficient methods for their detection and the application of these methods to samples from different sources. Four acid nucleic extraction techniques have been tested (classic proteinase K- phenol/chloroform, QIAamp Viral RNA Kit (Qiagen), SV Total RNA Isolation System (Promega) and NucleoSpin Virus L (Macherey-Nagel). Sensitivity and specificity of RT-PCR protocols for entero- (EV), hepatitis A (HAV) and small round structured (SRSV) viruses have been verified. Deep groundwater samples (100 L) were concentrated (2-step tangential flow ultrafiltration) and the concentrate contaminated with serial 10 fold dilutions of a known titre of poliovirus type 3. Extracted RNA was concentrated (microcon-100) and analysed by RT-PCR using specific EV primers and visualising amplification products by agarose gel electrophoresis. In addition, two different methods of RT-PCR for non-cultivable viruses have been tested: (a) RT-PCR and nested RT-PCR for HAV and (b) RT-PCR with generic primers and RT-PCR with specific primers for SRSV. Different specificity tests have been carried out in the presence of some of the commoner microorganisms. The most efficient, sensitive and specific protocols were used to test 35 x 100L deep groundwater samples. Sample concentrates were split with one part treated with chloroform and analysed by cell culture (BGM and Frp/3, derived from FrHK/4, cells) and the other tested by RT-PCR for HAV, EV and SRSV. Results demonstrated the high efficiency of the classic and QIAamp methods. Microcon-100 did not increase the sensitivity of the technique used. The highest sensitivity was observed for RT PCR with specific primers for SRSV and for nested RT-PCR for HAV. One sample showed a cytopathic effect, not confirmed at the third subculture, while the RT PCR allowed the detection of echovirus 7. Cell culture did not allow detection of the majority of the enteric viruses while PCR gave sensitive, specific and rapid detection of a range of agents in the same samples. Even if it was impossible to fix a virological quality standard, it would be necessary to find a viral indicator in order to achieve a complete preventive check which would be particularly useful in some cases (e.g. water never used before, after pollution accidents, for seasonal checking). PMID- 12639040 TI - A laboratory evaluation of the palatability of legumes to the field slug, Deroceras reticulatum Muller. AB - Slugs are major pests of many crops, including winter wheat, in temperate climates, yet current methods of control are often unreliable. The aim of this study is to investigate the potential for common legume species to act as an alternative source of food, or trap crop, for the most damaging agricultural pest species, the grey field slug, Deroceras reticulatum Muller, thereby reducing damage to the wheat crop. A series of three controlled-environment experiments were designed to assess this aim. Individual slugs were fed leaves of one of ten legume species together with winter wheat leaves for a 72-h period. A clear hierarchy of acceptability was shown, with red clover, lucerne, lupin and white clover showing significantly higher Acceptability Indices than the other six species tested. Red clover produced the greatest reduction in mean wheat consumption (78%) from day 1 to day 3. When species were fed individually, red clover was consumed in significantly greater quantities than any of the other treatments: 40% more than white clover and 56% more than wheat. Furthermore, when fed with red clover the amount of wheat consumed was some 50% less than when the latter was fed alone. The results indicate that legumes vary greatly in their acceptability to D. reticulatum and it is essential that a legume with a high Acceptability Index is chosen, which results in the least amount of wheat consumed. PMID- 12639041 TI - Determination of residues of endosulfan in human blood by a negative ion chemical ionization gas chromatographic/mass spectrometric method: impact of long-term aerial spray exposure. AB - A new and sensitive analytical method using negative ion chemical ionization gas chromatography/mass spectrometry in selective ion monitoring (SIM) mode has been developed for the determination of residues of endosulfan in the human blood. The residues of endosulfan are extracted from whole blood samples without separating the serum by the addition of 60% sulfuric acid at 10 degrees C followed by partition with hexane + acetone (9 + 1 by volume). The total endosulfan is quantified as the sum of alpha-endosulfan, beta-endosulfan and endosulfan sulfate in SIM mode. The mass-fragment ions used for this purpose that are monitored for in SIM mode include endosulfan diol: 95, 169, 214, 313, alpha-endosulfan: 99, 242, 270, 406, beta-endosulfan: 99, 242, 270, 406, and endosulfan sulfate: 97, 353, 386. Recovery experiments were conducted at the concentration range 1.0-100 pg ml(-1). Results showed 112-98% recovery of total endosulfan from the whole blood samples. The relative standard deviation was 1.49-2.68%. The method was found to be highly sensitive in quantifying endosulfan residues down to the 0.1 pg ml(-1) level. Conversion of endosulfan to endosulfan diol was found to be less than 0.1% under the conditions used. The results were compared with published data. The applications of the analytical method for the determination of endosulfan residues in real samples was tested by analyzing 106 human blood samples collected from a population living in Padre village, Kasargode District, Kerala, India, where aerial spraying of endosulfan has been a common agricultural practice over the years. The results showed that none of the blood samples contained residues of endosulfan (alpha-endosulfan 4 beta-endosulfan + endosulfan sulfate) or endosulfan diol. The results were confirmed by the detection of the appropriate amounts in a number of these samples which had subsequently been spiked with endosulfan. PMID- 12639042 TI - Monitoring of atrazine treatment on soil bacterial, fungal and atrazine-degrading communities by quantitative competitive PCR. AB - We report the development of quantitative competitive (QC) PCR assays for quantifying the 16S, 18S ribosomal and atzC genes in nucleic acids directly extracted from soil. QC-PCR assays were standardised, calibrated and evaluated with an experimental study aiming to evaluate the impact of atrazine application on soil microflora. Comparison of QC-PCR 16S and 18S results with those of soil microbial biomass showed that, following atrazine application, the microbial biomass was not affected and that the amount of 16S rDNA gene representing 'bacteria' increased transitorily, while the amount of 18S rDNA gene representing fungi decreased in soil. In addition, comparison of atzC QC-PCR results with those of atrazine mineralisation revealed that, in response to atrazine treatment, the amount of atzC gene increased transitorily in soil pre-treated with atrazine, suggesting that accelerated atrazine biodegradation in soil could be due to a transient increase in the size of the atrazine mineralising community. PMID- 12639043 TI - Learning performances of honeybees (Apis mellifera L) are differentially affected by imidacloprid according to the season. AB - To establish the sublethal concentrations domain, acute and chronic oral tests were conducted on caged honeybee workers (Apis mellifera L) using imidacloprid and a metabolite, 5-OH-imidacloprid, under laboratory conditions. The latter showed a 48-h oral LD50 value (153 ng per bee) five times higher than that of imidacloprid (30 ng per bee). Chronic feeding tests indicated that the lowest observed effect concentrations (LOEC) of imidacloprid and of 5-OH-imidacloprid on mortality of winter bees were 24 and 120 microg kg(-1) respectively. Behavioural effects of imidacloprid and 5-OH-imidacloprid were studied using the olfactory conditioning of proboscis extension response at two periods of the year. Winter bees surviving chronic treatment with imidacloprid and 5-OH-imidacloprid had reduced learning performances. The LOEC of imidacloprid was lower in summer bees (12 microg kg(-1)) than in winter bees (48 microg kg(-1)), which points to a greater sensitivity of honeybees behaviour in summer bees, compared to winter bees. PMID- 12639044 TI - Field relevance of a synergistic effect observed in the laboratory between an EBI fungicide and a chloronicotinyl insecticide in the honeybee (Apis mellifera L, Hymenoptera). AB - It had been found earlier that the chloronicotinyl insecticide thiacloprid (as the 480 g litre(-1) SC Calypso) poses a favourably low toxicity hazard to the honeybee, Apis mellifera L. As with pyrethroids, the metabolization of chloronicotinyl compounds involves monooxygenases, which are known to be inhibited by some ergosterol biosynthesis inhibitor (EBI) fungicides potentially co-applied with these insecticides. The potential synergistic enhancement of the toxicity of thiacloprid to honeybees when co-applied with such fungicides was therefore studied under laboratory and semi-field conditions. Fungicides of other chemical classes were also examined for synergistic potential to reveal other metabolic interactions. In the laboratory, only a slight synergistic effect was observed with the anilinopyrimidine fungicide examined, while a significant enhancement of thiacloprid toxicity to honeybees was found with EBI fungicides. In three tunnel tests conducted under different environmental conditions to simulate field exposure, no increased mortality was observed when honeybees were directly sprayed with thiacloprid (Calypso) alone or in combination with the EBI fungicide tebuconazole (250 g litre(-1) EW, Folicur). There was also no synergized reduction in the foraging intensity on the treated crop. In general, the foraging intensity decreased after thiacloprid treatment but was restored within 24-48 h. The hive vitality was not affected by either thiacloprid or its tank mix with tebuconazole. Our results suggest that, at the recommended use rates, thiacloprid poses a negligible lethal risk to honeybees when applied either alone or in tank mixes with fungicides of various chemical classes. PMID- 12639045 TI - Effects of cyazofamid against Plasmodiophora brassicae Woronin on Chinese cabbage. AB - Cyazofamid (4-chloro-2-cyano-N,N-dimethyl-5-p-tolylimidazole-1-sulfonamide) is a novel fungicide with high levels of activity against Oomycetes fungi and Plasmodiophora brassicae Woronin. The effects of cyazofamid were investigated against P. brassicae, the causal agent of clubroot disease in Chinese cabbage. Cyazofamid at 0.3 mg litre(-1) inhibited resting spore germination of this pathogen by about 80%. Cyazofamid at 3-10 mg litre(-1) exhibited fungicidal activity to resting spores of P. brassicae 1-10 days after treatment. When cyazofamid was applied to infested soil, both root-hair infections and club formation caused by P. brassicae were strongly inhibited at 1-3 mg kg(-1) dry soil. These results suggest that cyazofamid directly inhibits resting spore germination, thereby leading to the inhibition of root-hair infection and club formation. Cyazofamid at 3 mg kg(-1) dry soil also exhibited complete control of clubroot disease. The effect of broadcast soil application using a dust formulation at 2 kg AI ha(-1) (equivalent to 1.3 mg AI kg(-1) dry soil), and plug seedling tray application by a suspension concentrate formulation at 200 and 400 mg AI tray(-1) (30 x 60 x 4 cm3) against P. brassicae was also evaluated. Cyazofamid exhibited good efficacy against the pathogen. The sequential treatment including plug seedling tray application with cyazofamid and pre-plant broadcast soil application with the fungicide fluazinam also exhibited excellent levels of control. These results indicate that cyazofamid has a high potential to be an effective fungicide for the control of clubroot disease. PMID- 12639046 TI - Modulators of membrane drug transporters potentiate the activity of the DMI fungicide oxpoconazole against Botrytis cinerea. AB - Modulators known to reduce multidrug resistance in tumour cells were tested for their potency to synergize the fungitoxic activity of the fungicide oxpoconazole, a sterol demethylation inhibitor (DMI), against Botrytis cinerea Pers. Chlorpromazine, a phenothiazine compound known as a calmodulin antagonist, appeared the most potent compound. Tacrolimus, a macrolide compound with immunosuppressive activity, was also active. The synergism of chlorpromazine negatively correlated with the sensitivity of the parent strain and mutants of B. cinerea. The synergism was highest in a mutant that overexpressed the ATP-binding cassette transporter BcatrD, known to transport DMI fungicides such as oxpoconazole. The synergism of chlorpromazine positively correlated with its potency to enhance the accumulation of oxpoconazole in BcatrD mutants. These results indicate that chlorpromazine is a modulator of BcatrD activity in B. cinerea and suggest that mixtures of DMI fungicides with modulators may represent a perspective for the development of new resistance management strategies. PMID- 12639047 TI - Impact of alternative respiration and target-site mutations on responses of germinating conidia of Magnaporthe grisea to Qo-inhibiting fungicides. AB - Qo-inhibiting fungicides act as respiration inhibitors by binding to the Qo center of cytochrome b. Sensitivities of fungi to Qo inhibitors can be influenced by the induction of alternative respiration or by mutational changes of the cytochrome b target site. Previous studies on both mechanisms in Magnaporthe grisea (Hebert) Barr were focused on the mycelial stage of the pathogen. The present study describes the expression and impact of both resistance mechanisms during the stage of conidia germination. In the absence of a host, alternative respiration provided a >500-fold rescue from azoxystrobin during the germination of conidia derived from four wild-type isolates of M. grisea. This rescue potential during conidia gemination was substantially more pronounced than for mycelial growth. However, the pronounced effectiveness of alternative respiration during conidia germination was not apparent when barley leaves were protected with azoxystrobin prior to inoculation with conidia. In a comparison of a wild type strain and an alternative respiration-deficient mutant, azoxystrobin efficacies in suppressing symptom development differed by a factor of two, with full disease control achieved for both genotypes at 1 microg ml(-1) azoxystrobin. In contrast, conidia derived from two QoI-resistant target site mutants were highly resistant to azoxystrobin and trifloxystrobin and fully capable of infecting leaf surfaces protected with 10 microg ml(-1) of azoxystrobin. Both target-site mutants had emerged spontaneously in the presence of high azoxystrobin doses when residual mycelial growth was supported by alternative respiration. The effective silencing of alternative respiration in protective applications of Qo-inhibiting fungicides might constitute a strategy of slowing the emergence of highly resistant target site mutants. PMID- 12639048 TI - Evaluation of Erysiphe graminis f sp tritici field isolates for resistance to strobilurin fungicides with different SNP detection systems. AB - A single nucleotide polymorphism (snp) in the cytochrome b gene confers resistance to strobilurin fungicides in Erysiphe graminis DC fsp tritici Marchal. On the basis of this point mutation three different types of molecular markers have been developed. Cleaved amplified polymorphic sequences and allele-specific PCR were used to score resistant and sensitive isolates from specifically selected regional populations across Europe. The results of molecular tests were in total agreement with the resistance phenotypes revealed by in vivo tests. Serial dilutions of mixed samples (resistant/sensitive) delimited the detection for strobilurin-resistant alleles to a range of 10-50% for both marker classes. Due to these detection limits no mixture of mitochondria within individual isolates was found. Denaturing high performance chromatography was used to increase the detection sensitivity for the mutant allele. Although the detection limit was lowered to 5-10%, there was no evidence for the existence of mixed mitochondrial genotypes. PMID- 12639049 TI - Relative effects of the insecticide thiamethoxam on the predator Podisus nigrispinus and the tobacco whitefly Bemisia tabaci in nectaried and nectariless cotton. AB - The predaceous stinkbug Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) feeds on plants as well as on arthropod prey. The question arises whether feeding on plants might expose the predator to systemic insecticide via ingestion of the active ingredient or its metabolites through plant sap of treated plants. This interaction was investigated with nectaried and nectariless cotton plants cropped in pots and treated with the systemic insecticide thiamethoxam at 0.5, 1.0 and 2.0 mg per plant as a root drench. Development of P. nigrispinus fed on prey and on treated nectaried and nectariless cotton plants and confined at 15, 30 and 45 days after insecticide application, and adult reproduction from nymphs caged 30 days after treatment were determined. Podisus nigrispinus life history traits were unaffected by the type of cotton plant, nectaried or nectariless, but were significantly affected by insecticide dose and time after application. Developmental time was extended and fresh adult body weight was reduced by feeding on prey and treated plants. Nymphs caged on treated plants with the highest thiamethoxam concentration at 15 days after application produced only 13.2% of adults. Females emerged from nymphs caged on both plants and at all thiamethoxam concentrations at day 30 after application presented similar reproductive characteristics, except for age of first oviposition, which was delayed on plants treated with the highest thiamethoxam concentration. Thiamethoxam at 0.5 mg per plant restrained tobacco whitefly, Bemisia tabaci Gennadius (Homoptera: Aleyrodidae) colonization only during the first 15 days after application to either cotton plant, and similar immature densities were sampled at day 35 after application on treated and untreated plants. However, plants treated with 1.0 and 2.0 mg per plant as a drench and cropped in pots were protected from tobacco whitefly for up to 45 days after exposure to a whitefly colony. PMID- 12639050 TI - Laboratory studies on formation of bound residues and degradation of propiconazole in soils. AB - Laboratory studies on the formation of bound residues and on the degradation of the triazole fungicide propiconazole were conducted in two different soils. Soils treated with 14C-propiconazole were incubated at 22 degrees C and extracted exhaustively with a solvent at each sampling date until no further propiconazole was extracted. The solvent-extractable residues were used to measure propiconazole remaining in the soil, and the extracted soils were used to investigate bound residues of propiconazole. Mineralization of propiconazole was investigated by measuring [14C]carbon dioxide evolved from the soil samples. Formation of bound residues of propiconazole was higher in silty clay loam soil than in sandy loam soil, giving approximately 38 and 23% of the applied 14C, respectively. In contrast, the rates of degradation and mineralization of propiconazole were lower in silty clay loam soil than in sandy loam soil. Decreased extractability of the 14C residues with incubation time was observed with increased formation of bound residues. When the propiconazole remaining in the solvent-extractable residues was quantitatively measured by high-pressure liquid chromatographic analysis, the half-life value in sandy loam soil was about 315 days, while the half-life in silty clay loam soil exceeded the duration of the 1 year experimental period. Increased formation of bound residues was observed as propiconazole degraded with incubation time, suggesting that degradation products are involved in the formation of bound residues. Our study suggests that the formation of bound residues of propiconazole contributes to the persistence of this fungicide in soil. PMID- 12639051 TI - A mechanism for the formation of bis-glutathione conjugates of propargyl alcohol. AB - Our earlier research clearly revealed glutathione (GSH) conjugation as a major pathway for the metabolism of propargyl alcohol (2-propyn-1-ol) in rats and in mice. The identification of the metabolite 3,3-bis[(2-acetylamino-2 carboxyethyl)thio]-1-propanol (I) and its congeners represented the first example of multi-glutathione addition to a triple bond, and invoked further research to determine the mechanism for bis-conjugation. To determine whether GSH conjugated directly with propargyl alcohol or after oxidation of the latter to 2-propynal, urinary metabolites from rats administered deuterium-labeled propargyl alcohol were characterized. Following TLC separation and HPLC purification, mass spectrometry was used to show a single mass unit increase for metabolite I over that of the chemically synthesized standard. This result indicates that conjugation of propargyl alcohol with GSH to form the bis-conjugates occurred after initial oxidation to 2-propynal, a reaction that is analogous to a Michael addition. PMID- 12639053 TI - Biochemical study of resistance to imidacloprid in B biotype Bemisia tabaci from Guatemala. AB - Systemic uptake bioassays using excised cotton leaves confirmed resistance to imidacloprid in a Guatemalan population of the tobacco whitefly Bemisia tabaci Gennadius. Polyacrylamide gel electrophoresis of naphthyl esterases identified the insects as B-types. Upon collection from the field, resistance was determined to be 58-fold relative to a susceptible strain originating in the Imperial Valley of California. Resistance levels increased to 126-fold in this population during its continuous exposure to systemically treated cotton. In biochemical investigations, there was no detectable NADPH-dependent mixed function oxidase metabolism of 14C-imidacloprid at any time during the selection process. In contrast, microsomal preparations from housefly abdomens readily produced significant amounts of the mono-hydroxy and olefin derivatives of the parent compound. Detoxification of imidacloprid by housefly MFOs may account for reports of lower toxicity of the insecticide towards this insect compared with whiteflies, despite similar binding properties between imidacloprid and the nicotinic acetylcholine receptors in both species. PMID- 12639052 TI - Effect of the combined treatment of insecticides and an attractant for the control of Phloeotribus scarabaeoides, a pest of Olea europea. AB - Different insecticides have been tested for the control of the olive bark beetle, Phloeotribus scarabaeoides Bern. This scolytid can be managed at two points in its biological cycle: in pruned logs, where it excavates reproduction galleries, or in living trees, after emergence from the logs, where it digs feeding galleries. In mortality laboratory bioassays, the efficiency of organophosphorus insecticides has been ranked as follows: chlorpyrifos + dimethoate < formothion < methidathion. Formothion and methidathion, the two most efficient, were sprayed on olive logs together with a pyrethroid insecticide, deltamethrin, and a formulation which combined an organophosphorus (fenitrothion) and a pyrethroid (cypermethrin) insecticide. Deltamethrin inhibited the excavation of new reproduction galleries and induced a repellent effect on the olive pest. In contrast, none of the organophosphorus insecticides or the combination, fenitrothion + cypermethrin, were able to control the olive bark beetle. In olive trees, deltamethrin controlled this olive pest without showing the repellent effect observed for logs. Ethylene, a plant hormone, has been reported as an attractant for the olive bark beetle. The use of dispensers which released ethylene increased the number of P. scarabaeoides approaching the treated olive trees, thus favouring its use in a lure-and-trap control system. PMID- 12639054 TI - Toxicity of cyanogen to insects of stored grain. AB - Range-finding studies on the toxicity of cyanogen to all stages of five species of stored product Coleoptera are reported. The species were Rhyzopertha dominica (F), Sitophilus granarius (L), Sitophilus Oryzae (L), Tribolium castaneum (Herbst), Tribolium confusum Jacquelin du Val and Ephestia cautella (Walker). Exposures for 24 h to cyanogen at 1.3 mg litre(-1) controlled all external stages. Control of internal stages of Sitophilus species was achieved by a 5-day exposure to initial concentrations between 13.7 and 27.4 mg litre(-1), whereas R. dominica was controlled at 13.7 mg litre(-1). Cyanogen showed similar toxicity to all tested external stages and, in this respect, was more similar to methyl bromide than to phosphine. Its toxicity to insects increased with both relative humidity and concentration of carbon dioxide. Cyanogen was toxic to insects whether applied as a gas or in an aqueous solution. PMID- 12639055 TI - Wind tunnel studies on spray deposition on leaves of tree species used for windbreaks and exposure of honey bees. AB - A wind tunnel study was conducted to determine pesticide deposition on commonly used windbreak tree species used as spray drift barriers and associated exposure of honey bees. Although it has been known that windbreaks are effective in reducing chemical drift from agricultural applications, there is still an enormous information and data gap on details of the dependence of the mechanism on the biological materials of the barriers and on standardization of relevant assessment methods. Beneficial arthropods like honey bees are adversely affected by airborne drift of pesticides. A study was initiated by first establishing a wind tunnel to create a controlled environment for capture efficiency work. Suitable spray parameters were determined after a preliminary study to construct and develop a wind tunnel protocol. A tracer dye solution was sprayed onto the windbreak samples and honey bees located in the wind tunnel at various simulated wind speeds. Analysis of data from this work has shown that needle-like foliage of windbreak trees captures two to four times more spray than broad-leaves. In addition, it was determined that, at lower wind speeds, flying bees tend to capture slightly more spray than bees at rest. PMID- 12639056 TI - Development and preventative effect against pine wilt disease of a novel liquid formulation of emamectin benzoate. AB - Injection of the poorly water-soluble emamectin benzoate (EB) into pine trunks required the development of an efficient liquid formulation. For injection into big trees in forests a good rate of injection and a high active content were required. Tests on the viscosity and EB-solubilizing ability of 14 various solubilizers in diethylene glycol monobutyl ether (DGMBE) led to the selection of Sorpol SM-100PM as the solubilizer of the formulation. Relationships between the solubilizing ability and amounts of Sorpol SM-100PM and DGMBE relative to that of EB, and between the concentration of the latter and the viscosity or the injection rate of the formulation led to a novel 40 g litre(-1) emamectin benzoate formulation (Shot Wan Liquid Formulation), which was composed of EB (40), Sorpol SM-100PM (120), DGMBE (160) and distilled water (50 g litre(-1)) in methanol. Injection of this formulation at a dose of 10 g EB per unit volume of pine tree prevented over 90% of the trees from wilting caused by pine wood nematode, and this preventative effect continued for 3 years. Neither discolouration of the leaves nor injury around the injection hole on the trees was observed after injection of the formulation. PMID- 12639057 TI - Common names of pesticides recently approved by the BSI. PMID- 12639058 TI - Aspects of quality of life through end-stage renal disease. AB - This study was conducted to explore differentiated aspects of quality of life (QoL) throughout the course of end-stage renal disease (ESRD) with special regard to kidney transplantation (KTX). In detail two questions were addressed: (1) does social support change over time? and (2) is psychological distress a predictor of generic and disease-specific QoL? Two groups of ESRD-patients were investigated twice with different psychodiagnostic inventories. At t0 the Brief Symptom Inventory (BSI) and a validated German Social Support Questionnaire were administered. At t1 social support as well as generic and disease-specific QoL were investigated (SF-36, End-Stage Renal Disease Symptom Checklist Transplantation Module [ESRD-SCL]). Group A consisted of 80 patients treated by dialysis while on the waiting list for transplantation when investigated at t0 and was followed after successful transplantation at t1. Group B consisted of 222 patients investigated twice after successful KTX. Groups A and B did not differ with respect to clinical data and comorbidity, except for stronger immunosuppression in transplanted patients of group A. Group A demonstrated a statistically significant (5% explanation of variance) increased satisfaction with social support after KTX, whereas group B reported a decrease. The psychological distress at t0 predicted QoL at t1 in both groups (2-13% explanation of variance of SF-36, ESRD-SCL-scores). High psychological distress on dialysis and after KTX is a predictor for low long-term QoL. PMID- 12639059 TI - Validity of the St George's respiratory questionnaire at acute exacerbation of chronic bronchitis: comparison with the Nottingham health profile. AB - The purpose of the study is to compare a generic and a specific quality of life (QoL) instrument in the assessment of QoL in chronic bronchitis. Data from 320 patients were collected at acute exacerbation of chronic bronchitis (AECB), and from 230 patients during a subsequent stable phase (non-AECB), utilising both the specific St George's respiratory questionnaire (SGRQ) and the generic Nottingham health profile (NHP). Patients (maximum n = 200) reported significantly poorer QoL at AECB than at non-AECB for all domains except the SGRQ symptom domain (SRM = 0.02). The SGRQ was more sensitive than the NHP to QoL differences between patients. The correlations between the scores across the two assessments were generally higher for the SGRQ, with the correlations between the NHP and SGRQ being lower at AECB, suggesting that the instruments are measuring different constructs at AECB, with the SGRQ being less responsive to QoL change. Indeed, the NHP was generally more responsive to QoL change in terms of 'risk' and clinical factors, with the SGRQ symptom domain appearing particularly non responsive. In conclusion, this study suggests that the condition-specific SGRQ, and particularly its symptom domain, is less responsive than the generic NHP to QoL change accompanying AECB. This reflects the construct of the symptom domain which measures chronic bronchitis symptoms over the previous year. PMID- 12639060 TI - Interpretation of change and longitudinal validity of the quality of life for respiratory illness questionnaire (QoLRIQ) in inpatient pulmonary rehabilitation. AB - The quality of life for respiratory illness questionnaire (QoLRIQ) is an outcome measure for patients with asthma or chronic obstructive pulmonary disease (COPD). This study assessed the longitudinal validity, reliability of the change score and the interpretation of changes on the QoLRIQ in inpatient pulmonary rehabilitation, completed by 108 patients with moderate to severe asthma (39) or COPD (69). Domains and total score of the QoLRIQ changed significant (all p < 0.0002) with standardized response means from 0.46 to 0.90. All QoLRIQ-change scores were significantly correlated with self-rated change in health and in disease symptoms and with change in self-assessed health status (r from 0.2 to 0.61). There were several significant correlations between QoLRIQ-change scores and change in experienced invalidity, emotional well-being, anxiety, depressive symptoms and Rand-36-domains (r from 0.2 to 0.68). The intraclass correlation coefficient of change was 0.90. The size of a minimal important difference (MID), computed from a retrospective global rating of change by the patients and with the standard error of measurement, was 0.5 points on a 7-point response scale. Computation of the MID from retrospective assessment of change may not be valid because this change was significantly correlated to post-treatment health status and significantly higher than serial assessment of change. We conclude that the QoLRIQ is sensitive to change, longitudinally valid and reliable, with a MID of 0.5 points. These results enable the use of the QoLRIQ as an outcome measure in clinical trials with patients with moderate to severe asthma or COPD. The longitudinal measurement properties in less severe patients still need to be studied. PMID- 12639061 TI - Responsiveness of the quality of life in epilepsy inventory (QOLIE-89) in an antiepileptic drug trial. AB - This study examined relationships among responsiveness indices for health-related quality of life (HRQOL) measures administered to adults with epilepsy enrolled in an antiepileptic drug trial. The Quality of Life in Epilepsy (QOLIE)-89 was completed at baseline and at a 28-week follow-up. Six responsiveness indices (effect size (ES), standardized response mean (SRM), responsiveness statistic, paired t-test, area under receiver operating characteristic curve or ROC, F statistic) were calculated for each of the 16 QOLIE-89 scales, using two different external criteria for clinically significant change: (1) attainment of freedom from seizures with altered awareness, and (2) a two-category improvement between baseline and follow-up in a self-rating of the subject's overall condition. Spearman correlations among the six responsiveness indices for the 16 QOLIE-89 scales tended to be moderate to large (Spearman's p = 0.53-1.00; p's < 0.05 for 29 out of the 30 correlations). Rankings of the 16 scales across the two external criteria for change were similar for the responsiveness statistic (Spearman's p = 0.62; p < 0.05), but dissimilar for the other responsiveness indices (all p > 0.05). Both ES and SRM were well predicted by the other indices, except for ROC, using regression modeling. In conclusion, results using different responsiveness indices are comparable for a given external criterion. However, only the responsiveness statistic yielded robust results across two different external criteria. Responsiveness of this HRQOL measure can be reported in terms of previously established benchmarks for ESs, which can be predicted from other indices. PMID- 12639062 TI - How quality of life data contribute to our understanding of cancer patients' experiences? A study of patients with lung cancer. AB - A prospective study was conducted to measure quality of life in newly diagnosed lung cancer patients attending a chest clinic in a large teaching and district general hospital in a geographically defined area (northern sector of Glasgow, Scotland). Quality of life was assessed at two points in time, pre-diagnosis (baseline) and 3 months after diagnosis (follow-up) using three standard measures; the Nottingham Health Profile (NHP); the European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30), and its lung cancer supplement (QLQ-LC13). Out of 133 lung cancer patients diagnosed during the study period, 129 patients (97%) were interviewed pre diagnosis. Of these, only 63% of the patients had an active treatment. Ninety-six patients were alive at follow-up, of whom 82 patients were re-interviewed. Thus, only 82 patients who had complete data were used in the analysis. Comparing patients' pre-diagnosis and follow-up scores on the NHP, only sleep difficulties improved slightly. Patients reported increased perceived health problems of all other characteristics studied (energy, p = 0.0004; physical mobility, p = 0.0008). Similar results were observed on the EORTC questionnaires indicating that patients' functioning and global quality of life had decreased. The only significant improvement after 3 months was seen in patients' cough (p = 0.006). There were marked increases in hair loss (p < 0.0001), constipation (p = 0.007), and sore mouth (p = 0.0004). The findings suggest that patient-centred variables should receive sufficient consideration in the treatment of lung cancer. The study results clearly indicate that information on quality of life contributes to our understanding of patients' experiences of their cancer treatment. PMID- 12639063 TI - Measuring quality of palliative care: psychometric properties of the FAMCARE Scale. AB - This study measures quality of palliative care in 181 family members (i.e. spouse, child) to cancer victims with terminal disease 1 month after the time of death. The specific aim was to explore the underlying factor structure and dimensionality of the 20 items of the FAMCARE Scale, measuring family satisfaction with health care given to the patient and to them. The results from a factor analysis, a Mokken Scaling Program analysis, and a reliability analysis, showed that 19 out of the 20 items form a strong one-dimensional scale. Since the scale is one-dimensional, the possibility of reducing the number of items should be explored in future research. Our recommendation is to measure satisfaction with care 1-2 months after the death of the patient. One should also explore the possibilities of measuring satisfaction with care prospectively as an integral part of the palliative care program. PMID- 12639064 TI - Evaluating podiatry services: testing a treatment specific measure of health status. AB - This study reports on the preliminary testing of a new measure designed for use alongside EQ-5D in evaluating outcomes in podiatry: the Podiatry Health Questionnaire (PHQ). Individuals aged 18 years or more, receiving podiatry services in clinic or domicilliary locations across four NHS Trusts in Yorkshire and Humberside UK took part in a questionnaire survey. Respondents reported high levels of problems on all six PHQ dimensions. Correlations suggested that the PHQ and EQ-5D were measuring distinct constructs. The levels on each dimension were well defined in terms of self-rated morbidity on the PHQ visual analogue scale (PHQvas) and the EQ-5Dvas, although PHQvas appeared to be slightly more sensitive to changes in health on the dimensions. There was a strong relationship between clinicians' Podiatry Clinical Score rating and reported symptoms for four out of six PHQ dimensions and PHQvas. The PHQ was able to distinguish respondents in terms of their self-reported morbidity in EQ-5D and in terms of their morbidity as assessed by clinicians. It is suggested that the respondent completed PHQ appears to be a useful new measure for assessing foot-related health. However, further investigation of the psychometric properties of the measure is required. PMID- 12639065 TI - Unidimensionality and reliability under Mokken scaling of the Dutch language version of the SF-36. AB - The sub-scales of the SF-36 in the Dutch National Study are investigated with respect to unidimensionality and reliability. It is argued that these properties deserve separate treatment. For unidimensionality we use a non-parametric model from item response theory, called the Mokken scaling model, and compute the corresponding scalability coefficients. We estimate reliability under the Mokken model, assuming that the items are double homogeneous, and compare it to Cronbach's alpha. The scalability of the sub-scale general health perceptions is medium (H = 0.46), and for the other sub-scales it is strong (H > or = 0.6). The reliability in terms of alpha indicates that all sub-scales can be used in basic research (alpha > 0.70), but that only physical functioning can be used for clinical applications of quality of life (alpha > 0.90). The relative merits of our approach are discussed. PMID- 12639066 TI - Health-related quality of life in patients with coronary artery disease treated for angina: validity and reliability of German translations of two specific questionnaires. AB - The German versions of two patient-perceived heart disease specific health related quality of life (HRQL) questionnaires, the Seattle Angina Questionnaire (SAQ) and the MacNew Heart Disease questionnaire, were examined for their psychometric properties in patients with angiographically documented coronary artery disease and angina who were treated either medically or invasively and followed up for 1 year. Both HRQL questionnaires and the modified Canadian Cardiovascular Society (CCS) angina-associated disability scale were completed by 158 patients at baseline and 12 months later when they also completed a generic health status questionnaire, the SF-36. Both specific HRQL questionnaires were acceptable to patients. Three of the four MacNew scales, but none of the SAQ scales, discriminated between patients by baseline CCS disability levels I and IV. Internal consistency ranged from 0.75 to 0.94 for the SAQ and from 0.86 to 0.97 for the MacNew scales. Test-retest reliability over a 4-week period of time ranged from 0.45 to 0.81 for the SAQ scales and 0.61 to 0.68 for the MacNew scales. Over 12 months, HRQL improved (p < 0.001) on three of the five SAQ and on all four of the MacNew scales with the responsiveness statistic ranging from 0.59 to 1.55 for the SAQ and 0.86 to 1.12 for the MacNew. The 12 month scores on all SAQ and MacNew scales were significantly higher in patients who improved than those who deteriorated on the SF-36 reported health transition question. We conclude that the SAQ and the MacNew are both valid, reliable, and responsive in German, that the MacNew discriminates better between angina grades at baseline, that HRQL improves over 12 months with both measures, that the SAQ angina frequency and disease perception scales have the largest effect sizes, and that the 12-month change in HRQL with both instruments was associated with change in SF-36 reported health transition status. PMID- 12639067 TI - Applying exercise stage of change to a low-income underserved population. AB - OBJECTIVES: To validate the transtheoretical model for exercise behavior and the constructs of decisional balance and self-efficacy for exercise in a low-income, poorly educated primary care sample. METHODS: Patients attending public primary care clinics from 4 separate sites in Louisiana were interviewed regarding their health behaviors. RESULTS: The data provide equivocal support for applying the transtheoretical model for exercise and integrating it with other models of behavior change within this population. CONCLUSIONS: Further studies modifying the decisional balance measures are necessary before definitive statements regarding the applicability of these models to exercise within this specialized population can be made. PMID- 12639068 TI - Parental monitoring and adolescent alcohol risk in a clinic population. AB - OBJECTIVES: To assess the association between parental monitoring and adolescent alcohol risk. METHODS: Adolescent managed-care patients completed a survey of their involvement with alcohol and their perceptions of monitoring and other forms of parental influence. RESULTS: Frequent monitoring was associated with less likelihood that adolescents had engaged in a variety of alcohol-risk behaviors or had been in a variety of risk situations. Choosing to share alcohol related concerns with other adults and placing importance on parents' opinions were not as consistently related to alcohol risk. CONCLUSIONS: The need to enhance parental monitoring as a proactive protective parental response is indicated. PMID- 12639069 TI - Conducting web-based survey research: a lesson in internet designs. AB - OBJECTIVES: To examine the tacit knowledge resulting from designing a complex Web site to facilitate administration of an Internet-based survey. METHODS: Formative methods guided studying the feasibility of conducting an online survey using versions of the Keirsey Temperament Sorter II and the National College Health Risk Behavior Survey. RESULTS: Web-site design software and pilot testing were critical to success. Incorporating the target audience as consumers in both Web site development and formative evaluation simplified data collection and analysis. CONCLUSIONS: Online instrumentation used in survey research is both practical and desirable. Future refinements are necessary, but improved designs can emanate from this particular effort. PMID- 12639070 TI - Skin penetration operators' knowledge and attitudes towards infection control. AB - OBJECTIVES: To assess the knowledge and attitudes of owners/managers of commercial skin-penetration premises regarding infection control. METHODS: A telephone survey was conducted with a randomly selected sample of 874 owners/managers. RESULTS: Participants appeared to lack knowledge of essential infection-control practices. Less than 39% correctly identified recommended disinfection procedures, and between 12% to 67% were not aware of inappropriate sterlization procedures. Almost all participants accepted the need for guidelines. Half acknowledged a need to improve their infection-control compliance, and most accepted having their premises regularly checked by the councils. CONCLUSIONS: There is a considerable opportunity to increase infection control compliance among skin-penetration operators. PMID- 12639071 TI - Focus groups with school staff to explore asset-building innovativeness. AB - OBJECTIVE: To assist with interpretations of findings from a survey assessing the diffusion of an asset-building innovation in 3 school districts. METHODS: Eight focus groups were conducted with 69 school administrators, teachers, and guidance counselors in 3 school districts. Three reviewers examined transcripts for common themes. RESULTS: The themes that arose related to effective communication channels, barriers that prevent diffusion in the schools, the need for improved relationships, and addressing of students' needs. CONCLUSIONS: The study illustrates the advantages of linking qualitative and quantitative methods when performing evaluations by enhancing both the validity and usefulness of findings. PMID- 12639072 TI - Differences in the use of a school mental health program in secondary schools. AB - OBJECTIVE: To examine the use of a school-based mental health program that works to prevent drug and alcohol problems and promote student mental health. METHODS: Students' presenting concerns, services used, service format and people to whom students were referred for services were investigated. Student use among schools was compared using chi-square analyses. RESULTS: The program was highly used, with a range of concerns differentiated by school type. Student concerns, service format, and student referral and follow-up varied significantly. CONCLUSION: Student use is a product of student need, school structure, and mental health educator. PMID- 12639073 TI - A multiple case study of neighborhood partnerships for positive youth development. AB - OBJECTIVE: To identify factors associated with successful neighborhood mobilization to prevent teen pregnancy using a positive youth development approach. METHODS: A multiple case study was conducted on 5 neighborhood partnerships using in-person interviews with key informants, observation of meetings, and review of existing documents. RESULTS: Competent staff, strong sense of community, support of key organizations, shared leadership, effective group process, and a perceived need for the initiative influenced level of mobilization. CONCLUSION: Organizing neighborhoods around teen pregnancy prevention, even using a youth development approach, is challenging but can be accomplished when the right constellation of factors is in place. PMID- 12639074 TI - Suggestions for using the web to collect data. PMID- 12639076 TI - Implantable cardioverter defibrillators in primary and secondary prevention: a systematic review of randomized, controlled trials. AB - BACKGROUND: Sudden cardiac death is common in persons with cardiovascular disease. PURPOSE: To assess the efficacy of implantable cardioverter defibrillators (ICDs) in persons at increased risk for sudden cardiac death. DATA SOURCES: MEDLINE (1980-2002), EMBASE (1980-2002), Cochrane Controlled Clinical Trial Registry (2002, Volume 3), other databases, and conference proceedings. Primary study authors and device manufacturers were contacted, and bibliographies of relevant papers were hand searched. STUDY SELECTION: Randomized, controlled clinical trials evaluating ICDs versus usual care were selected. DATA EXTRACTION: Two reviewers extracted data independently. DATA SYNTHESIS: Eight trials were included in the final analysis (4909 patients, 1154 deaths). Compared with usual care (most commonly amiodarone therapy), ICDs significantly reduced sudden cardiac death (relative risk [RR], 0.43 [95% CI, 0.35 to 0.53]) and all-cause mortality (RR, 0.74 [CI, 0.67 to 0.82]). The included trials were divided a priori into two categories: secondary prevention (involving patients resuscitated after cardiac arrest or unstable ventricular tachycardia or ventricular fibrillation [ n = 1963]) and primary prevention (involving patients at increased risk for sudden cardiac death but without documented cardiac arrest, ventricular fibrillation, or ventricular tachycardia [ n = 2946]). Regardless of baseline risk, ICDs were equally efficacious in preventing sudden cardiac death in both types of trials (RR, 0.50 [CI, 0.38 to 0.66] for secondary prevention vs. 0.37 [CI, 0.27 to 0.50] for primary prevention). However, the magnitude of benefit in total mortality varied within the primary prevention trials depending on baseline risk for sudden cardiac death. CONCLUSIONS: Implantable cardioverter defibrillators prevent sudden cardiac death regardless of baseline risk. However, their impact on total mortality is sensitive to baseline risk for arrhythmic death. Decisions about resource allocation for ICDs depend on accurate stratification of patients according to risk. PMID- 12639077 TI - High prevalence of anal human papillomavirus infection and anal cancer precursors among HIV-infected persons in the absence of anal intercourse. AB - BACKGROUND: Anal cancer and its precursor lesion, anal squamous intraepithelial lesions (SILs), are associated with human papillomavirus (HPV) infection. Anal HPV infection and anal SIL are common in HIV-positive men who have sex with men; receptive anal intercourse is presumed to be the mode of acquisition of HPV. OBJECTIVE: To assess the prevalence and risk factors for anal HPV infection and anal SIL in HIV-positive men with no history of anal intercourse. DESIGN: Cross sectional study. SETTING: Hopital Europeen Georges Pompidou outpatient clinic, Paris, France. PATIENTS: 118 HIV-infected men. MEASUREMENTS: 50 HIV-positive heterosexual male injection drug users with no history of anal intercourse and 67 HIV-infected men who had sex with men were evaluated by using anal cytologic, anal histologic, and anal HPV DNA testing. RESULTS: 23 of the 50 heterosexual injection drug users (46%) had anal HPV infection. Low-grade SIL (LSIL) was found in 8 patients (16%) and high-grade SIL (HSIL) in 9 patients (18%). Among the 67 men who had sex with men, anal HPV infection was found in 57 patients (85%), LSIL in 33 patients (49%), and HSIL in 12 patients (18%). In univariate analysis, risk factors for abnormal anal cytologic or histologic findings in injection drug users included CD4+ cell counts less than 250 x 10(6) cells/L (odds ratio, 5.7 [95% CI, 1.6 to 20.4]), plasma HIV RNA viral load greater than 1.7 log copies/mL (odds ratio, 8.9 [CI, 1.1 to 76.0]), previous AIDS-defining event (odds ratio, 4.3 [CI, 1.2 to 15.6]), and anal HPV detection (odds ratio, 5.7 [CI, 1.6 to 20.4]). Risk factors among men who had sex with men included having more than 10 lifetime receptive anal intercourse episodes (odds ratio, 5.6 [CI, 1.6 to 19.8]) and anal HPV detection (odds ratio, 8.7 [CI, 1.9 to 39.0]). CONCLUSIONS: Anal HPV infection and anal SIL may be acquired in the absence of anal intercourse in HIV positive men. The prevalence of HSIL is high among HIV-positive injection drug users. All HIV-positive men with CD4+ cell counts less than 500 x 10(6) cells/L, regardless of history of anal intercourse, should be considered for anal cytologic screening; however, additional studies are needed to determine the efficacy of this procedure to prevent anal cancer in these populations. PMID- 12639078 TI - The impact of protein intake on renal function decline in women with normal renal function or mild renal insufficiency. AB - BACKGROUND: In individuals with moderate to severe renal insufficiency, low protein intake may slow renal function decline. However, the long-term impact of protein intake on renal function in persons with normal renal function or mild renal insufficiency is unknown. OBJECTIVE: To determine whether protein intake influences the rate of renal function change in women over an 11-year period. DESIGN: Prospective cohort study. SETTING: Nurses' Health Study. PARTICIPANTS: 1624 women enrolled in the Nurses' Health Study who were 42 to 68 years of age in 1989 and gave blood samples in 1989 and 2000. Ninety-eight percent of women were white, and 1% were African American. MEASUREMENTS: Protein intake was measured in 1990 and 1994 by using a semi-quantitative food-frequency questionnaire. Creatinine concentration was used to estimate glomerular filtration rate (GFR) and creatinine clearance. RESULTS: In multivariate linear regression analyses, high protein intake was not significantly associated with change in estimated GFR in women with normal renal function (defined as an estimated GFR > or = 80 mL/min per 1.73 m2). Change in estimated GFR in this subgroup over the 11-year period was 0.25 mL/min per 1.73 m2 (95% CI, -0.78 to 1.28 mL/min per 1.73 m2) per 10-g increase in protein intake; the change in estimated GFR was 1.14 mL/min per 1.73 m2 (CI, -3.63 to 5.92 mL/min per 1.73 m2) after measurement-error adjustment for protein intake. In women with mild renal insufficiency (defined as an estimated GFR > 55 mL/min per 1.73 m2 but <80 mL/min per 1.73 m2), protein intake was significantly associated with a change in estimated GFR of -1.69 mL/min per 1.73 m2 (CI, -2.93 to -0.45 mL/min per 1.73 m2) per 10-g increase in protein intake. After measurement-error adjustment, the change in estimated GFR was -7.72 mL/min per 1.73 m2 (CI, -15.52 to 0.08 mL/min per 1.73 m2) per 10-g increase in protein intake, an association of borderline statistical significance. High intake of nondairy animal protein in women with mild renal insufficiency was associated with a significantly greater change in estimated GFR (-1.21 mL/min per 1.73 m2 [CI, -2.34 to -0.33 mL/min per 1.73 m2] per 10-g increase in nondairy animal protein intake). CONCLUSIONS: High protein intake was not associated with renal function decline in women with normal renal function. However, high total protein intake, particularly high intake of nondairy animal protein, may accelerate renal function decline in women with mild renal insufficiency. PMID- 12639079 TI - The relative safety of ephedra compared with other herbal products. AB - BACKGROUND: Ephedra is widely used in dietary supplements that are marketed to promote weight loss or increase energy; however, the safety of this product has been questioned because of numerous case reports of adverse events. OBJECTIVE: To compare the risk for adverse events attributable to ephedra and other herbal products. DESIGN: Comparative case series. SETTING: American Association of Poison Control Centers Toxic Event Surveillance System Database Annual Report, 2001. MEASUREMENTS: The relative risk and 95% confidence interval for experiencing an adverse reaction after ephedra use compared with other herbs. This risk was defined as the ratio of adverse reactions to ephedra versus other products, divided by the ratio of their relative use in the United States. RESULTS: Products containing ephedra accounted for 64% of all adverse reactions to herbs in the United States, yet these products represented only 0.82% of herbal product sales. The relative risks for an adverse reaction in persons using ephedra compared with other herbs were extremely high, ranging from 100 (95% CI, 83 to 140) for kava to 720 (CI, 520 to 1100) for Ginkgo biloba. CONCLUSIONS: Ephedra use is associated with a greatly increased risk for adverse reactions compared with other herbs, and its use should be restricted. PMID- 12639081 TI - The mini-CEX: a method for assessing clinical skills. AB - OBJECTIVE: To evaluate the mini-clinical evaluation exercise (mini-CEX), which assesses the clinical skills of residents. DESIGN: Observational study and psychometric assessment of the mini-CEX. SETTING: 21 internal medicine training programs. PARTICIPANTS: Data from 1228 mini-CEX encounters involving 421 residents and 316 evaluators. INTERVENTION: The encounters were assessed for the type of visit, sex and complexity of the patient, when the encounter occurred, length of the encounter, ratings provided, and the satisfaction of the examiners. Using this information, we determined the overall average ratings for residents in all categories, the reliability of the mini-CEX scores, and the effects of the characteristics of the patients and encounters. MEASUREMENTS: Interviewing skills, physical examination, professionalism, clinical judgment, counseling, organization and efficiency, and overall competence were evaluated. RESULTS: Residents were assessed in various clinical settings with a diverse set of patient problems. Residents received the lowest ratings in the physical examination and the highest ratings in professionalism. Comparisons over the first year of training showed statistically significant improvement in all aspects of competence, and the method generated reliable ratings. CONCLUSIONS: The measurement characteristics of the mini-CEX are similar to those of other performance assessments, such as standardized patients. Unlike these assessments, the difficulty of the examination will vary with the patients that a resident encounters. This effect is mitigated to a degree by the examiners, who slightly overcompensate for patient difficulty, and by the fact that each resident interacts with several patients. Furthermore, the mini-CEX has higher fidelity than these formats, permits evaluation based on a much broader set of clinical settings and patient problems, and is administered on site. PMID- 12639082 TI - Update in cardiology. PMID- 12639080 TI - Support for physician deception of insurance companies among a sample of Philadelphia residents. AB - BACKGROUND: Some physicians seem to be willing to sanction deception of insurance companies. Little is known about public attitudes regarding this practice. OBJECTIVE: To assess public attitudes regarding physician deception of insurance companies. DESIGN: Cross-sectional survey using clinical vignettes. SETTING: Philadelphia County Courthouse, Philadelphia, Pennsylvania. PARTICIPANTS: Convenience sample of 700 prospective jurors. MEASUREMENTS: Participants were asked whether, in response to restriction of health care, a physician should 1) accept restriction, 2) appeal restriction, or 3) misrepresent a patient's condition to obtain the desired service. The proportion of respondents reporting that the physician should misrepresent a patient's condition was determined. RESULTS: 26% of respondents sanctioned deception, 70% supported appealing, and 4% supported accepting the insurance company decision. Among the 27% of respondents believing physicians have inadequate time to appeal coverage decisions, 50% sanctioned deception. CONCLUSIONS: Sanctioning of deception was substantial in this sample of prospective jurors. Preferences regarding insurance company deception are related to perceptions of physician workload and may further pressure physicians struggling to balance advocacy with honesty. PMID- 12639083 TI - Evaluation of 21st-century risks of smallpox vaccination and policy options. AB - The United States stopped vaccinating against smallpox in 1972 because the risks were judged to outweigh the benefits. The possibility of a terrorist attack using smallpox has led to renewed interest in a vaccination program. Smallpox vaccination carries considerable risks, which may be of greater concern today than in the late 1960s because of the increased prevalence of immunosuppression and atopy in the population. This paper reviews the clinical presentations of major adverse events after vaccination and the rates of occurrence of these events observed in the 1960s. The normal dynamics of the spread of smallpox is slow, and usually only persons who have had close personal contact with an overtly ill patient are affected. There are several preattack vaccination policy options, but immunization of medical workers, especially those who might have close contact with infected patients, is sufficient in the absence of a known threat of a bioterrorist attack or the identification of a smallpox-infected person. PMID- 12639084 TI - Prevention of ventilator-associated pneumonia: an evidence-based systematic review. AB - BACKGROUND: Ventilator-associated pneumonia is a common cause of morbidity in critically ill patients. Interventions beneficial to the prevention of ventilator associated pneumonia would therefore have a significant impact on the care of these patients. PURPOSE: To perform a literature review and synthesis of methods for prevention of ventilator-associated pneumonia. DATA SOURCES: MEDLINE (1966 2001), the Cochrane Library, and bibliographies of retrieved articles. STUDY SELECTION: Studies were required to be prospective and controlled in design and to evaluate clinically important or surrogate outcomes. Surrogate outcomes were required to have a direct link to clinically important outcomes supported by the literature. DATA EXTRACTION: Data on patients, definitions, study design, and outcomes were abstracted and graded by using preestablished criteria. DATA SYNTHESIS: The preventive practices with the strongest supportive evidence were semi-recumbent positioning, sucralfate instead of H2-antagonists for stress ulcer prophylaxis, and selective digestive tract decontamination. Aspiration of subglottic secretions and oscillating beds may be useful in select populations. There is no evidence to support specific methods of enteral feeding or increased frequency of ventilator circuitry changes. CONCLUSIONS: After evaluation of potential benefits and risks, the authors recommend considering several specific interventions to reduce the incidence of ventilator-associated pneumonia: semi recumbent positioning in all eligible patients, sucralfate rather than H2 antagonists in patients at low to moderate risk for gastrointestinal tract bleeding, and aspiration of subglottic secretions and oscillating beds in select patient populations. Selective digestive tract decontamination is not recommended because routine use may increase antimicrobial resistance. PMID- 12639085 TI - A new paradigm for the treatment of sepsis: is it time to consider combination therapy? AB - Despite the advances in supportive care and the availability of potent antimicrobial agents, mortality from sepsis, a leading cause of death in intensive care units, has not improved. Over the last decade, clinical trials with numerous adjunctive therapies, including antiendotoxin antibodies and inhibitors of the inflammatory response, have yielded disappointing results. Recently, treatment with recombinant human activated protein C reduced mortality 6% compared with controls. Given the likelihood that many processes in the complex pathophysiology of sepsis are simultaneously activated, it is unlikely that therapy directed at any one of them, as has been done in the past, will dramatically improve survival. Rather, a combination of therapies directed at many arms of the septic process, much like the strategy used for cancer and HIV infection, is required. Given the likelihood that sepsis represents an excessive innate immune response to microbial products, vigorous attempts must be made to develop rapid assays that reflect the level of innate immune activation. Such assays could be used to identify patients who would benefit from therapy and to monitor their response so that overtreatment does not completely abrogate host defense mechanisms and render these patients susceptible to fatal infection. It is now time to test a new therapeutic paradigm based on an improved understanding of the pathophysiology of the septic process and the recognition that we may have reached the limits of adjunctive monotherapy. PMID- 12639086 TI - Cardiac events in patients undergoing noncardiac surgery: shifting the paradigm from noninvasive risk stratification to therapy. AB - Internists and cardiologists are often asked to estimate the risk for perioperative myocardial infarction or cardiac death in patients being considered for noncardiac surgery. Estimating this risk in an individual patient is difficult and complex. Although noninvasive imaging tests are often used for this purpose, a review of the literature reveals that the positive predictive value of noninvasive imaging tests is uniformly low and that they do not provide information beyond that obtained by assessing simple clinical risk variables. Moreover, no evidence exists that noninvasive imaging tests lead to a therapeutic strategy that reduces the risk for perioperative myocardial infarction or cardiac death. Since the publication of guidelines for preoperative risk stratification by the American College of Cardiology/American Heart Association in 1996 and the American College of Physicians in 1997, three clinical trials have shown that beta-blocker therapy reduces the risk for perioperative cardiac events. This paper focuses on the relationship between risk stratification and subsequent therapy to minimize or eliminate risk. In short, the paradigm is shifting from predicting which patient is at high risk for having a perioperative cardiac event to minimizing the likelihood of such an event with specific perioperative pharmacologic therapy. PMID- 12639087 TI - The clinical use of implantable cardioverter defibrillators: where are we now? Where should we go? PMID- 12639088 TI - Them and us. PMID- 12639089 TI - Tests of glycemia. PMID- 12639091 TI - The wrong patient. PMID- 12639092 TI - The wrong patient. PMID- 12639093 TI - The wrong patient. PMID- 12639095 TI - Bad outcomes of questionable medical decisions. PMID- 12639096 TI - Bad outcomes of questionable medical decisions. PMID- 12639098 TI - Bad outcomes of questionable medical decisions. PMID- 12639100 TI - Myiasis in a traveler: a moving story. PMID- 12639099 TI - A novel defect in interferon-gamma secretion in patients with refractory nontuberculous pulmonary mycobacteriosis. PMID- 12639101 TI - Hypercalcemia after 1,25-dihydroxyvitamin D3 production in an end-stage kidney. PMID- 12639102 TI - Summaries for patients. Ventilator-associated pneumonia. PMID- 12639103 TI - Summaries for patients. Opinions about doctors deceiving insurance companies to obtain health services for their patients. PMID- 12639104 TI - Summaries for patients. Ephedra is associated with more adverse effects than other herbs. PMID- 12639105 TI - Summaries for patients. The effect of dietary protein intake on kidney function in women with normal or mildly abnormal kidneys. PMID- 12639106 TI - Summaries for patients. Anal cancer precursors in persons with HIV infection. PMID- 12639107 TI - Summaries for patients. Who benefits from implantable heart defibrillators? PMID- 12639108 TI - Direct observation of fine structure transitions in a paramagnetic nickel(II) complex using far-infrared magnetic spectroscopy: a new method for studying high spin transition metal complexes. AB - Novel far-infrared (FIR) absorption spectroscopy in conjunction with multiple, fixed external magnetic fields (FIR magnetic spectroscopy, FIRMS) has been used to investigate pseudotetrahedral complexes with the formula M(PPh(3))(2)Cl(2) (M = Ni, Zn; Ph = C(6)H(5)). Crystal structures have been reported for the Ni complex; we report the structure of the Zn complex. Transmission spectra at 5 K of Ni(PPh(3))(2)Cl(2) (S = 1) at zero magnetic field exhibit absorption bands at 11.41, 15.28, and 23.0 cm(-1). The two lower frequency bands show great sensitivity to external magnetic field, and their field dependence is as expected for electron spin transitions allowing precise determination of the following parameters: |D| = 13.35(1) cm(-1), |E| = 1.93(1) cm(-1), g(x,y) = 2.20(1), g(z) = 2.00(1). Corresponding spectra of Zn(PPh(3))(2)Cl(2) (S = 0) exhibit bands only at >20 cm(-1), which show no field dependence. FIRMS is a promising technique for direct investigation of the electronic structure of high-spin transition metal complexes. PMID- 12639109 TI - First radical cation salt of paramagnetic transition metal complex containing TTF as ligand, [Cu(II)(hfac)2(TTF-py)2](PF6) x 2CH2Cl2 (hfac = hexafluoroacetylacetonate and TTF-py = 4-(2-tetrathiafulvalenyl ethenyl)pyridine). AB - The preparation, X-ray crystal structure, EPR data, and magnetic measurement of [Cu(II)(hfac)(2)(TTF-py)(2)](PF(6)).2CH(2)Cl(2), a novel material where the conducting and the localized spin systems are covalently linked through conjugated bridges, are reported. The partial oxidation of the TTF-type organic donor ligand yielded the first radical cation salt of a paramagnetic transition metal complex. Moreover, this compound shows a mixed valence state at the unimolecular level, and additionally, the arrangement of the molecules in the crystal structure revealed the presence of isolated mixed valence TTF dimers. PMID- 12639110 TI - Synthesis and characterization of an open framework ballium selenide: Ga4Se7(en)2 x (enH)2. AB - An open framework gallium selenide, Ga(4)Se(7)(en)(2).(enH)(2), has been prepared by the direct reaction of gallium (Ga) and selenium (Se) in ethylenediamine (en), in which both covalent and hydrogen bonds have been employed to combine the inorganic structures and organic spacers to build layers with micropores. Its structure has been determined by X-ray diffraction. Its thermal and optical properties have been characterized by TGA and UV-vis, Raman, and IR spectroscopies, respectively. PMID- 12639111 TI - Synthesis and Structure of KBGe2O6: the first chiral zeotype borogermanate with 7 ring channels. AB - Among the known zeolite topologies, odd rings are rare with the exception of 5 ring. We report here the hydrothermal synthesis and structural characterization of a novel compound, KBGe(2)O(6), the first germanate-based zeotype material with 7-ring channels. It crystallized in the orthorhombic system, space group P2(1)2(1)2(1) (No. 19), a = 4.8037(3) A, b = 10.2063(7) A, c = 10.7402(10) A, V = 526.57(5) A(3), Z = 3. The framework topology of this compound is previous unknown with the vertex symbol 4.6.4.6.6.7(2)(vertex1), 4.6.4.6.6.7(2)(vertex2), and 6.6.6.6.6(2) x 7(2)(vertex3). It is worth noting that the structure of the compound is chiral containing helices condensed from GeO(4) tetrahedra, which arrange around 2(1) screw axes. Furthermore, the K(+) ions within the channels are mobile and can be partially ion-exchanged with Na(+) at room temperature. PMID- 12639112 TI - Reversible binding of dioxygen by the copper(I) complex with tris(2 dimethylaminoethyl)amine (Me6tren) ligand. AB - At low temperatures, the mononuclear copper(I) complex of the tetradentate tripodal aliphatic amine Me(6)tren (Me(6)tren = tris(2-dimethylaminoethyl)amine) [Cu(I)(Me(6)tren)(RCN)](+) first reversibly binds dioxygen to form a 1:1 Cu-O(2) species which further reacts reversibly with a second [Cu(I)(Me(6)tren)(RCN)](+) ion to form the dinuclear 2:1 Cu(2)O(2) adduct. The reaction can be observed using low temperature stopped-flow techniques. The copper superoxo complex as well as the peroxo complex were characterized by resonance Raman spectroscopy. The spectral characteristics and full kinetic and thermodynamic results for the reaction of [Cu(I)(Me(6)tren)(RCN)](+) with dioxygen are reported. PMID- 12639113 TI - Copper(I)-dioxygen reactivity of [(L)Cu(I)](+) (L = tris(2-pyridylmethyl)amine): kinetic/thermodynamic and spectroscopic studies concerning the formation of Cu-O2 and Cu2-O2 adducts as a function of solvent medium and 4-pyridyl ligand substituent variations. AB - The kinetic and thermodynamic behavior of O(2)-binding to Cu(I) complexes can provide fundamental understanding of copper(I)/dioxygen chemistry, which is of interest in chemical and biological systems. Here we report stopped-flow kinetic investigations of the oxygenation reactions of a series of tetradentate copper(I) complexes [(L(R))Cu(I)(MeCN)](+) (1(R), R=H, Me, tBu, MeO, Me(2)N) in propionitrile (EtCN), tetrahydrofuran (THF), and acetone. The syntheses of 4 pyridyl substituted tris(2-pyridylmethyl)amine ligands (L(R)) and copper(I) complexes are detailed. Variations of ligand electronic properties are manifested in the electrochemistry of 1(R) and nu(CO) of [(L(R))Cu(I)-CO](+) complexes. The kinetic studies in EtCN and THF show that the O(2)-reactions of 1(R) follow the reaction mechanism established for oxygenation of 1(H) in EtCN (J. Am. Chem. Soc. 1993, 115, 9506), involving reversible formation (k(1)/k(-1)) of [(L(R))Cu(II)(O(2-))](+) (2(R)), which further reacts (k(2)/k(-2)) with 1(R) to form the 2:1 Cu(2)O(2) complex [[(L(R))Cu(II)](2)(O(2)(2-))](2+) (3(R)). In EtCN, the rate constants for formation of 2(R) (k(1)) are not dramatically affected by the ligand electronic variations. For R = Me and tBu, the kinetic and thermodynamic parameters are very similar to those of the parent complex (1(H)); e.g., k(1) is in the range 1.2 x 10(4) to 3.1 x 10(4) M(-1) s(-1) at 183 K. With the stronger donors R = MeO and Me(2)N, more significant effects were observed, with the expected increase in thermodynamic stability of resultant 2(R) and 3(R) complexes, and decreased dissociation rates. The modest ligand electronic effects manifested in EtCN are due to the competitive binding of solvent and dioxygen to the copper centers. In THF, a weakly coordinating solvent, the formation rate for 2(H) is much faster (>/=100 times) than that in EtCN, and the thermodynamic stabilities of both the 1:1 (K(1)) and 2:1 (beta = K(1)K(2)) copper-dioxygen species are much higher than those in EtCN (e.g., for 2(H), deltaH(o) (K(1))=-41 kJ mol(-1) in THF versus -29.8 kJ mol(-1) in EtCN; for 3(H), deltaH(o) (beta)=-94 kJ mol(-1) in THF versus -77 kJ mol(-1) in EtCN). In addition, a more significant ligand electronic effect is seen for the oxygenation reactions of 1(MeO) in THF compared to that in EtCN; the thermal stability of superoxo- and peroxocopper complexes are considerably enhanced using L(MeO) compared to L(H). In acetone as solvent, a different reaction mechanism involving dimeric copper(I) species [(L(R))(2)Cu(I)(2)](2+) is proposed for the oxygenation reactions, supported by kinetic analyses, electrical conductivity measurements, and variable-temperature NMR spectroscopic studies. The present study is the first systematic study investigating both solvent medium and ligand electronic effects in reactions forming copper-dioxygen adducts. PMID- 12639114 TI - Synthesis and spectroelectrochemical studies of mixed heteroleptic chelate complexes of ruthenium(II) with 1,8-bis(2-pyridyl)-3,6-dithiaoctane (pdto) and substituted 1,10-phenanthrolines. AB - Reaction of dichlorotris(triphenylphosphine) ruthenium(II) [RuCl(2)(PPh(3))(3)] with 1,8-bis(2-pyridyl)-3,6-dithiaoctane (pdto), a (N(2)S(2)) tetradentate donor, yields a new compound [Ru(pdto)(PPh(3))Cl]Cl (1), which has been fully characterized. (1)H and (31)P NMR studies of 1 in acetonitrile at several temperatures show the substitution of both coordinated chloride and triphenylphosphine with two molecules of acetonitrile, as confirmed by the isolation of the complex [Ru(pdto)(CH(3)CN)(2)]Cl(2) (2). Cyclic voltammetric and spectroelectrochemical techniques allowed us to determine the electrochemical behavior of compound 1. The substitution of the chloride and triphenylphosphine by acetonitrile molecules in the Ru(II) coordination sphere of compound 1 was also established by electrochemical studies. The easy substitution of this complex led us to use it as starting material to synthesize the substituted phenanthroline coordination compounds with (pdto) and ruthenium(II), [Ru(pdto)(4,7-diphenyl-1,10-phenanthroline)]Cl(2).4H(2)O (3), [Ru(pdto)(1,10 phenanthroline)]Cl(2).5H(2)O (4), [Ru(pdto)(5,6-dimethyl-1,10 phenanthroline)]Cl(2).5H(2)O (5), [Ru(pdto)(4,7-dimethyl-1,10 phenanthroline)]Cl(2).3H(2)O (6), and [Ru(pdto)(3,4,7,8-tetramethyl-1,10 phenanthroline)]Cl(2).4H(2)O (7). These compounds were fully characterized, and the crystal structure of 4 was obtained. Cyclic voltammetric and spectroelectrochemical techniques allowed us to determine their electrochemical behavior. The electrochemical oxidation processes in these compounds are related to the oxidation of ionic chlorides, and to the reversible transformation from Ru(II) to Ru(III). On the other hand, a single reduction process is associated to the reduction of the substituted phenanthroline in the coordination compound. The E(1/2) (phen/phen(-)) and E(1/2) (Ru(II)/Ru(III)) for the compounds (3-7) were evaluated, and, as expected, the modification of the substituted 1,10 phenanthrolines in the complexes also modifies the redox potentials. Correlations of both electrochemical potentials with pK(a) of the free 1,10-phenathrolines, lambda(max) MLCT transition band, and chemical shifts of phenanthrolines in these complexes were found, possibly as a consequence of the change in the electron density of the Ru(II) and the coordinated phenanthroline. PMID- 12639115 TI - Synthesis and structure of the metallic K6Tl17: a layered tetrahedral star structure related to that of Cr3Si. AB - The title compound, the Tl-richest in the K-Tl system, has been synthesized in Ta containers via direct reaction of the elements at 400 degrees C followed by quenching to room temperature and subsequent annealing at 150 degrees C for 4 weeks. It crystallizes in the orthorhombic space group Cccm (No. 66) with a = 16.625(1) A, b = 23.594(2) A, c = 15.369(2) A (22 degrees C), and Z = 8. Two different Tl(12) units consisting of augmented tetrahedral stars are condensed into layers of such tetrahedra, and further Tl(2) dumbbells and the potassium cations also interconnect the stars and layers into a three-dimensional network. The former anionic Tl(8) subunits clearly resemble those in the heteroatomic 3-D structure of cubic Cr(3)Si before their augmentation with bridging atoms. The compound is metallic (rho(270) = 22.6 micro omega x cm, alpha = 0.0023 K(-)(1)) and shows Pauli-like paramagnetic susceptibility (chi(296) = 1.1 x 10(-4) emu/mol). EHTB calculations illustrate the importance of Tl p-orbital bonding, the positive Tl-Tl overlap populations up to E(F), and greater strengths of the Tl-Tl bonding between and about the surface of the augmented Tl(12) units. Cations between the thallium layers play specific and important roles in the structure. PMID- 12639116 TI - Half-sandwich titanium(IV) complexes with Klaui's tripod ligand. AB - Treatment of [Ti(O-i-Pr)(2)Cl(2)] with NaL(OEt) (L(OEt)(-) = [CpCo[P(O)(OEt)(2)](3)](-), Cp = eta(5)-C(5)H(5)) afforded [L(OEt)Ti(O-i Pr)(2)Cl] that reacted with HCl in ether to give [L(OEt)TiCl(3)] (1). The average Ti-O and Ti-Cl distances in 1 are 1.975 and 2.293 A, respectively. Reaction of titanyl sulfate with NaL(OEt) in water followed by addition of HBF(4) afforded [L(OEt)TiF(3)] (2), the Ti-O and Ti-F distances of which are 2.020(2) and 1.792(2) A, respectively. The Zr(IV) analogue [L(OEt)ZrF(3)] (3) was prepared similarly from zirconyl nitrate, NaL(Oet), and HBF(4) in water. The Zr-O and average Zr-F distances in 3 are 2.139(2) and 1.938(2) A, respectively. Treatment of 1 with tetrachlorocatechol (H(2)Cl(4)cat) afforded [L(OEt)Ti(Cl(4)cat)Cl] (4). The average Ti-O(P), Ti-O(C), and Ti-Cl distances in 4 are 1.972, 1.926, and 2.334 A, respectively. Hydrolysis of 4 in the presence of Et(3)N yielded the mu oxo dimer [(L(OEt))(2)Ti(2)(Cl(4)cat)(2)(mu-O)] (5). The average Ti-O(P), Ti O(C), and Ti-O(Ti) distances in 5 are 2.027, 1.926, and 1.7977(9) A. Treatment of 1 with 1,1'-binaphthol (BINOLH(2)) in the presence of Et(3)N afforded [(L(OEt))(2)Ti(2)(mu-O)(2)(mu-BINOL)] x 2BINOLH(2) (6.2BINOLH(2)). Complex 1 is capable of catalyzing ring opening of epoxides with Me(3)SiN(3) under solvent free conditions presumably via a Ti-azide intermediate. PMID- 12639117 TI - Synthesis, crystal structures, and magnetic properties of two cyano-bridged tungstate(V)-manganese(II) bimetallic magnets. AB - The reaction of manganese acetate with octacyanotungastate in an aqueous solution of concentrated acetic acid gives two new three-dimensional cyano-bridged manganese(II)-tungstate(V) bimetallic assemblies, [Mn(II)(2)(H(2)O)(2)(CH(3)COO)][W(V)(CN)(8)].2H(2)O (1) (tetragonal space group I4/mcm, a = b = 11.9628(9) A, c = 13.367(2) A, and Z = 4) and Cs(I)(0.5)Mn(II)(2)[W(V)(CN)(8)](CH(3)CO(2))(1.5).H(2)O (2) (monoclinic space group C2/c, a = 16.274(2) A, b = 22.948(6) A, c = 13.196(1) A, beta = 128.040(6) degrees, and Z = 8). In complex 1, W(V)(CN)(8) adopts a square antiprismatic geometry, and each CN group coordinates to the Mn(II) ions forming W-Mn(4)-W Mn(4)-...columnar linkages where four sites on the Mn(II) ion with octahedral geometry are occupied by CN groups. The columns are parallel and interlock, yielding a network structure. Complex 2 contains two different coordination geometries for W(V)(CN)(8), namely, square antiprismatic and dodecahedral. The columnar structures appear also in 2, where the Mn(II) ions in two different environments provide three and four coordinated sites to the CN groups. The columns are bridged by both dodecahedral W(V)(CN)(8) groups and acetates. Cs ions were intercalated in the lattice by the formation of short attractive contacts with the acetates. The field-cooled magnetization, ac susceptibility, and the field dependence of magnetization measurements show that both 1 and 2 are ferrimagnets with ordering temperatures 40 and 45 K, respectively. The investigation of the magnetostructural correlation shows that the ferrimagnetic ordering in 1 and 2 are attributed to the dominant antiferromagnetic exchange pathways d(z)2(W)-d(x')(y')(Mn) and d(x)2(-y)2(W) - d(x')(y')(Mn). PMID- 12639118 TI - Dioxo-bridged dinuclear manganese(III) and -(IV) complexes of pyridyl donor tripod ligands: combined effects of steric substitution and chelate ring size variations on structural, spectroscopic, and electrochemical properties. AB - The syntheses and structural, spectral, and electrochemical characterization of the dioxo-bridged dinuclear Mn(III) complexes [LMn(mo-O)(2)MnL](ClO(4))(2), of the tripodal ligands tris(6-methyl-2-pyridylmethyl)amine (L(1)) and bis(6-methyl 2-pyridylmethyl)(2-(2-pyridyl)ethyl)amine (L(2)), and the Mn(II) complex of bis(2 (2-pyridyl)ethyl)(6-methyl-2-pyridylmethyl)amine (L(3)) are described. Addition of aqueous H(2)O(2) to methanol solutions of the Mn(II) complexes of L(1) and L(2) produced green solutions in a fast reaction from which subsequently precipitated brown solids of the dioxo-bridged dinuclear complexes 1 and 2, respectively, which have the general formula [LMn(III)(mu O)(2)Mn(III)L](ClO(4))(2). Addition of 30% aqueous H(2)O(2) to the methanol solution of the Mn(II) complex of L(3) ([Mn(II)L(3)(CH(3)CN)(H(2)O)](ClO(4))(2) (3)) showed a very sluggish change gradually precipitating an insoluble black gummy solid, but no dioxo-bridged manganese complex is produced. By contrast, the Mn(II) complex of the ligand bis(2-(2-pyridyl)ethyl)(2-pyridylmethyl)amine (L(3a)) has been reported to react with aqueous H(2)O(2) to form the dioxo bridged Mn(III)Mn(IV) complex. In cyclic voltammetric experiments in acetonitrile solution, complex 1 shows two reversible peaks at E(1/2) = 0.87 and 1.70 V (vs Ag/AgCl) assigned to the Mn(III)(2) <--> Mn(III)Mn(IV) and the Mn(III)Mn(IV) <--> Mn(IV)(2) processes, respectively. Complex 2 also shows two reversible peaks, one at E(1/2) = 0.78 V and a second peak at E(1/2) = 1.58 V (vs Ag/AgCl) assigned to the Mn(III)(2) <--> Mn(III)Mn(IV) and Mn(III)Mn(IV) <--> Mn(IV)(2) redox processes, respectively. These potentials are the highest so far observed for the dioxo-bridged dinuclear manganese complexes of the type of tripodal ligands used here. The bulk electrolytic oxidation of complexes 1 and 2, at a controlled anodic potential of 1.98 V (vs Ag/AgCl), produced the green Mn(IV)(2) complexes that have been spectrally characterized. The Mn(II) complex of L(3) shows a quasi reversible peak at an anodic potential of E(p,a) of 1.96 V (vs Ag/AgCl) assigned to the oxidation Mn(II) to Mn(III) complex. It is about 0.17 V higher than the E(p,a) of the Mn(II) complex of L(3a). The higher oxidation potential is attributable to the steric effect of the methyl substituent at the 6-position of the pyridyl donor of L(3). PMID- 12639119 TI - Ruthenium(III) polyaminocarboxylate complexes: efficient and effective nitric oxide scavengers. AB - The preparation of two Ru(III) polyaminocarboxylate complexes, AMD6245 and AMD6221, and their nitrosyl analogues, AMD6204, AMD6263, and AMD3689, is described. The compounds are characterized by IR, ES-MS, and (13)C and (15)N NMR spectroscopy where appropriate and cyclic voltammetry. The crystal structures for AMD6245, AMD6263, and AMD3689 are presented. AMD6245 (C(10)H(14)N(2)O(9)Ru) crystallized in the P2(1)/c space group with a = 8.4382(2) A, b = 8.8304(2) A, c = 17.6321(4) A, beta = 99.603(o), V = 1295.3(2) A(3), and Z = 4. AMD6263 (C(10)H(14)N(3)O(10)Ru) crystallized in the P2(1)/c space group with a = 9.9043(4) A, b = 13.1144(3) A, c = 12.0914(4) A, beta = 100.191(o), V = 1545.8(5) A(3), and Z = 4. AMD3689 (C(14)H(24.56)N(4)O(13.28)Ru) crystallized in the P1 space group with a = 8.838(2) A, b = 9.452(3) A, c = 13.419(4) A, alpha = 78.413(6)(o), beta = 75.804(6)(o), gamma = 73.562(6)(o), V = 1031.8(5) A(3), and Z = 2. The reaction of AMD6245 and AMD6221 with nitric oxide is investigated using EPR spectroscopy and stopped flow kinetics. Upon reaction with NO, a linear, diamagnetic [RuNO](6) complex is formed. The substitution reaction of AMD6245 with NO proceeds with a second-order rate constant of 2.24 x 10(7) M(-1) s(-1) at 7.3 degrees C (pH = 7.4; 50 mM phosphate buffer). The substitution reaction of AMD6221 with NO proceeds with a second-order rate constant of 3 x 10(5) M(-1) s(-1) at 20 degrees C (pH = 7.4; 50 mM phosphate buffered saline). The NO scavenging ability was assessed using a RAW264 murine macrophage assay by measuring the difference in nitrite produced between untreated control cells and treated cells. At 100 microM AMD6245 has [NO(2-)] = 12.5 microM less than the untreated cells and AMD6221 has [NO(2-)] = 37.6 microM less than the untreated cells. There is an insignificant difference in the amount of nitrite produced between AMD6263 or AMD3689 treated cells and untreated cells. PMID- 12639120 TI - Zinc(II)-templated synthesis of a [2]-catenane consisting of a 2,2',6',2' ' terpyridine-incorporating cycle and a 1,10-phenanthroline-containing ring. AB - A nonsymmetrical [2]-catenane has been synthesized, with a 5-coordinated metal center (Zn(2+)) as template. One of the two rings contains a terdentate ligand (2,2',6',2' '-terpyridine) and the other one incorporates a bidentate chelate (1,10-phenanthroline). The first ring was prepared separately and, subsequently, Zn(2+) was used as the gathering and threading element to pass the stringlike component through the ring. This open-chain species bears two terminal olefins, which were reacted with Grubbs first-generation catalyst (ring-closing metathesis) to afford the desired catenane. Hydrogenation of the double bond and removal of the zinc(II) template afforded the final free [2]-catenane in 40% yield from the terdentate ligand-containing cycle and the diolefinic compound. Complexation studies on this new pentacoordinating catenane were carried out with Fe(II) or Cu(II). The most interesting observation is that the 5-coordinated complexes obtained are strongly stabilized. Their electrochemical reduction occurs at negative potentials. PMID- 12639121 TI - Kinetics of oxygen exchange between the two isomers of bisulfite ion, disulfite Ion (S(2)O(5)(2-)), and water as studied by oxygen-17 nuclear magnetic resonance spectroscopy. AB - The nuclear magnetic transverse relaxation time of oxygen-17 in aqueous sodium bisulfite solutions in the pH range from 2.5 to 5 was measured over a range of temperatures, pH, and S(IV) concentrations at an ionic strength of 1.0 m. From these data the rate law for oxygen exchange between bisulfite ion and water was determined and found to be consistent with oxygen exchange occurring via the reactions SHO(3)(-) + H(+) SO(2) + H(2)O, SO(3)H(-) + SHO(3-) SO(3)(2-) + SO(2) + H(2)O, and SO(3)H(-) + SHO(3-) S(2)O(5)(2-) + H(2)O, where the symbol SHO(3-) refers to both isomeric forms of bisulfite ion, one in which the hydrogen is bonded to the sulfur (denoted HSO(3-)) and another in which the hydrogen is bonded to an oxygen atom (denoted SO(3)H(-)). The SO(3)H(-) isomer exchanges oxygen atoms with water much more rapidly than does the HSO(3-) isomer. The value of k(-1) was determined and is in essential agreement with the results of a previous determination by relaxation measurements. The value of k(16a) + k(16b) was also found, and k(16b) is at least as large as k(16a). The rate and mechanism of oxygen exchange between the two bisulfite ion environments were studied by analyzing the broadening of the HSO(3-) resonance. Oxygen exchange occurs through isomerization caused by proton transfers. PMID- 12639122 TI - Synthesis, X-ray crystal structure, and redox and electronic properties of iron(III)-polyimidazole complexes relevant to the metal sites of iron proteins. AB - A new tripod N(3) ligand (L), containing three imidazole rings, was synthesized in good yield. At variance with usual aromatic ligands with N(2) or N(3) donor sets such as pyridine or pyrazole derivatives, L stabilizes the Fe(III) oxidation state. The corresponding iron(III) complexes [Fe(L)Cl(3)] (1) and [Fe(L)(2)](ClO(4))(3) (2) were prepared and characterized by X-ray structural analysis and spectroscopic methods. The coordination environment around all the Fe(III) centers has a distorted octahedral geometry. [Fe(L)Cl(3)] (1) belongs to the monoclinic system, space group P2(1)/n, a = 9.7406(5) A, b = 17.207(2) A, c = 14.615(2) A, beta = 104.448(9)(o) Z = 4, V = 2372.1(4) A(3); R = 0.044, R(w) = 0.055. [Fe(L)(2)](ClO(4))(3) (2) belongs to the monoclinic system, space group P2(1)/c, a = 16.1057(15) A, b = 11.1079(12) A, c = 26.283(2) A, beta = 102.062(10)(o), Z = 4, V = 4598.2(8) A(3); R = 0.0465, R(w) = 0.0902. The Fe N((i)PrIm) bond lengths are systematically longer than the Fe-N(MeIm) ones. Compound 2 is a highly anisotropic low-spin Fe(III) complex displaying a rather unusual EPR spectrum with a sharp signal at g = 3.5 and a broad one at g approximately 1.6. The fitting of this EPR spectrum is discussed. PMID- 12639123 TI - Spectroscopic and thermal investigations of the fluoroaluminate complex formation in NaF-KF and LiF-NaF-KF eutectics. AB - The dissolution and complex formation of fluoroaluminates in two eutectic alkalifluoride mixtures, NaF-KF (FNAK) and LiF-NaF-KF (FLINAK), have been investigated by Raman, NMR, and thermal analysis. Melting and dissolution took place stepwise. The eutectic alkalifluoride mixtures with minor amounts of dissolved fluoroaluminate salts started melting at around 460 and 740 degrees C for FLINAK and FNAK mixtures, respectively. Total melting/dissolution of mixtures with 9-11 mol % aluminum fluoro salts added took place near 780 degrees C in the FLINAK solvent and at approximately 900 degrees C for FNAK solutions. The solidified melts were characterized by Raman bands at 561 (nu(1)), 391 (nu(2)), and 328 cm(-1) (nu(5)) and a (27)Al NMR chemical shift near 0 ppm originating from isolated AlF(6)(3-) octahedral ions. The Raman and NMR signals due to AlF(6)(3-) were also observed at temperatures where the samples were only partly melted. Upon total melting, a pronounced dissociation of AlF(6)(3-) into AlF(5)(2 ) and fluoride ions took place. At even higher temperatures, the equilibrium was displaced in favor of AlF(5)(2-) in the FNAK solvent. The AlF(5)(2-) ion was characterized by an intensive Raman band at 558 cm(-1) and an increasingly positive (27)Al chemical shift with raising temperature, e.g., of 16 ppm at 935 degrees C. PMID- 12639124 TI - Electronic structure of ReO3Me by variable photon energy photoelectron spectroscopy, absorption spectroscopy and density functional calculations. AB - Valence photoelectron (PE) spectra have been measured for ReO(3)Me using a synchrotron source for photon energies ranging between 20 and 110 eV. Derived branching ratios (BR) and relative partial photoionization cross sections (RPPICS) are interpreted in the context of a bonding model calculated using density functional theory (DFT). Agreement between calculated and observed ionization energies (IE) is excellent. The 5d character of the orbitals correlates with the 5p --> 5d resonances of the associated RPPICS; these resonances commence around 47 eV. Bands with 5d character also show a RPPICS maximum at 35 eV. The RPPICS associated with the totally symmetric 4a(1) orbital, which has s-like character, shows an additional shape resonance with an onset of 43 eV. The PE spectrum of the inner valence and core region measured with photon energies of 108 and 210 eV shows ionization associated with C 2s, O 2s, and Re 4f and 5p electrons. Absorption spectra measured in the region of the O1s edge showed structure assignable to excitation to the low lying empty "d" orbitals of this d(0) molecule. The separation of the absorption bands corresponded with the calculated orbital splitting and their intensity with the calculated O 2p character. Broad bands associated with Re 4d absorption were assigned to (2)D(5/2) and (2)D(3/2) hole states. Structure was observed associated with the C1s edge but instrumental factors prevented firm assignment. At the Re 5p edge, structure was observed on the (2)P(3/2) absorption band resulting from excitation to the empty "d" levels. The intensity ratios differed from that of the O 1s edge structure but were in good agreement with the calculated 5d character of these orbitals. An absorption was observed at 45 eV, which, in the light of the resonance in the 4a(1) RPPICS, is assigned to a 4a(1) --> ne, na(2) transition. The electronic structure established for ReO(3)Me differs substantially from that of TiCl(3)Me and accounts for the difference in chemical behavior found for the two complexes. PMID- 12639125 TI - Solid-state chelation of metal ions by ethylenediaminetetraacetate intercalated in a layered double hydroxide. AB - The solid-state chelation of transition metal ions (Co(2+), Ni(2+), and Cu(2+)) from aqueous solutions into the lithium aluminum layered double hydroxide ([LiAl(2)(OH)(6)]Cl x 0.5H(2)O or LDH) which has been pre-intercalated with EDTA (ethylenediaminetetraacetate) ligand has been investigated. The intercalated metal cations form [M(edta)](2)(-) complexes between the LDH layers as indicated by elemental analysis, powder X-ray diffraction, and IR and UV-vis spectroscopies. If metal chloride or nitrate salts are used in the reaction with the LDH then co-intercalation of either the Cl(-) or NO(3)(-) anions is observed. In the case of metal acetate salts the cations intercalate without the accompanying anion. This can be explained by the different intercalation selectivity of the anions in relation to the LDH. In the latter case the introduction of the positive charge into LDH structure was compensated for by the release from the solid of the equivalent quantity of lithium and hydrogen cations. Time-resolved in-situ X-ray diffraction measurements have revealed that the chelation/intercalation reactions proceed very quickly. The rate of the reaction found for nickel acetate depends on concentration as approximately k[Ni(Ac)(2)](3). PMID- 12639126 TI - Syntheses and structures of photochromic silver(I) coordination polymers with cis 1,2-dicyano-1,2-bis(2,4,5-trimethyl-3-thienyl)ethene. AB - Five novel silver(I) coordination polymers with cis-1,2-dicyano-1,2-bis(2,4,5 trimethyl-3- thienyl)ethene (cis-dbe) were synthesized and are characterized in this paper. Treatment of AgCF(3)SO(3) or AgCF(3)CO(2) with cis-dbe afforded [Ag(2)(cis-dbe)(CF(3)SO(3))(2)] (1) and [Ag(2)(cis-dbe)(CF(3)CO(2))(2)] (2), and both complexes exhibit a 1-D infinite chain structure with two cyano groups and two thienyl groups of the ligand bridging four metal ions. Reaction of AgC(n)()F(2)(n)(+1)CO(2) with cis-dbe gave rise to an unprecedented cocrystallization of a 2-D sheet structure, [Ag(2)(cis dbe)(C(n)F(2)(n)(+1)CO(2))(2)], where n = 2 (3), 3 (4), and 4 (5). Upon irradiation with 450 nm light, these five silver(I) complexes turned orange or red from yellow, and the color reverted to yellow on exposure to 560 nm light, indicative of the reversible cyclization/ring-opening reaction occurring in the crystalline phase. Furthermore, different anions gave not only the different structural dimensions but also the different photoresponsive patterns. The correlation between the crystal structures and the photochromic reactivity is discussed. PMID- 12639127 TI - Synthesis and characterization of optically active and racemic forms of cyclometalated Rh(III) complexes. An experimental and theoretical emission study. AB - The optically active cyclometalated Rh(III) complexes, delta[Rh(thpy4,5p(R,R)py)(2)TAP]Cl, lambda[Rh(thpy4,5p(S,S)py)(2)TAP]Cl, and delta[Rh(phpy4,5p(R,R)py)(2)TAP]Cl (where TAP = 1,4,5,8-tetraazaphenanthrene, thpy4,5p(R,R)py = (8R,10R)-2-(2'-thienyl)-4,5-pinenopyridine and phpy4,5p(R,R)py = (8R,10R)-2-(2'-phenyl)-4,5-pinenopyridine) have been prepared and characterized. Their photophysics has been examined in parallel with that of rac[Rh(thpy)(2)TAP]Cl and rac[Rh(phpy)(2)TAP]Cl. Their behaviors have been rationalized from results of TD-DFT calculations. The complexes with thienylpyridine (thpy) as cyclometalating ligands exhibit (3)CT (from thpy to TAP) and (3)LC(pi-pi) (centered on thpy) emissions in a solvent matrix at 77 K and one (3)CT luminescence at room temperature. In contrast, with phenylpyridine (phpy), the complexes show only one (3)CT emission (from phpy to TAP) at both temperatures. PMID- 12639128 TI - Electrochemical and spectral studies on the reductive nitrosylation of water soluble iron porphyrin. AB - The reaction of iron(III) (meso-tetrakis(N-methylpyridinium-4-yl)porphyrin (Fe(III)TMPyP) with nitric oxide (NO) was studied by electronic absorption spectroscopy, ESR, and electrochemical and spectroelectrochemical techniques in aqueous solutions with pH from 2.2 to 12.0. Fe(III)TMPyP has been found to undergo a reductive nitrosylation in all pHs, and the product of nitric oxide binding to the porphyrin has been determined as iron(II) porphyrin nitrosyl complex ([Fe(II)(NO)TMPyP]). The rate of the reductive nitrosylation exhibits a tendency to get faster with increase in pH. An intermediate species was observed around neutral pH by spectroelectrochemical technique and was proposed to be the iron(II) nitrosyl complex of the mu-oxo dimeric form of FeTMPyP, which is known to be a predominant in neutral solutions. PMID- 12639129 TI - Synthesis, structure, and dissociation equilibrium of [Co(eta5-C5H5)(Se2C6H4)]2, a novel metalladiselenolene complex. AB - A (phenylenediselenolato)cobalt complex dimer, [Co(eta(5) C(5)H(5))(Se(2)C(6)H(4))](2) (1), was synthesized by a reaction of carbonyl(eta(5)-cyclopentadienyl)diiodocobalt(III) ([Co(eta(5) C(5)H(5))I(2)(CO)]) with poly(o-diselenobenzene). The structure of 1, determined by single-crystal X-ray crystallography, was found to be located in the space group of P2(1)/c (No. 14), with a = 9.3346(5) A, b = 11.6477(9) A, c = 10.2179(5) A, beta = 111.491(1) degrees, and Z = 2. Covalent Co-Se bonds bridge the metal centers. In solution, dimers and monomers coexist at equilibrium. The dissociation equilibrium constant of 1 in solution was evaluated by (1)H NMR spectra at several temperatures between 20 and 80 degrees C. Dissociation enthalpies/entropies were found to be 50/110, 60/120, and 88 kJ mol(-1)/200 J K( 1) mol(-1) in dimethyl sulfoxide-d(6), benzene-d(6), and chloroform-d(1), respectively. PMID- 12639130 TI - Self-assembly in palladium(II) and platinum(II) chemistry: the biomimetic approach. AB - The self-assembly of complex cationic structures by combination of cis-blocked square planar palladium(II) or platinum(II) units with bis(pyridyl) ligands having bridging amide units has been investigated. The reactions have yielded dimers, molecular triangles, and polymers depending primarily on the geometry of the bis(pyridyl) ligand. In many cases, the molecular units are further organized in the solid state through hydrogen bonding between amide units or between amide units and anions. The molecular triangle [Pt(3)(bu(2)bipy)(3)(mu-1)(3)](6+), M = Pd or Pt, bu(2)bipy = 4,4'-di-tert-butyl-2,2'-bipyridine, and 1 = N-(4 pyridinyl)isonicotinamide, stacks to give dimers by intertriangle NH.OC hydrogen bonding. The binuclear ring complexes [[Pd(LL)(mu-2)](2)](CF(3)SO(3))(4), LL = dppm = Ph(2)PCH(2)PPh(2) or dppp = Ph(2)P(CH(2))(3)PPh(2) and 2 = NC(5)H(4)-3 CH(2)NHCOCONHCH(2)-3-C(5)H(4)N, form transannular hydrogen bonds between the bridging ligands. The complexes [[Pd(LL)(mu-3)](2)](CF(3)SO(3))(4), LL = dppm or dppp, L = PPh(3), and 3 = N,N'-bis(pyridin-3-yl)-pyridine-2,6-dicarboxamide, and [[Pd(LL)(mu-4)](2)](CF(3)SO(3))(4), LL = dppm, dppp, or bu(2)bipy, L = PPh(3), and 4 = N,N'-bis(pyridin-4-yl)-pyridine-2,6-dicarboxamide, are suggested to exist as U-shaped or square dimers, respectively. The ligands N,N'-bis(pyridin-3 yl)isophthalamide, 5, or N,N'-bis(pyridin-4-yl)isophthalamide, 6, give the complexes [[Pd(LL)(mu-5)](2)](CF(3)SO(3))(4) or [[Pd(LL)(mu 6)](2)](CF(3)SO(3))(4), but when LL = dppm or dppp, the zigzag polymers [[Pd(LL)(mu-6)](x)](CF(3)SO(3))(2)(x) are formed. When LL = dppp, a structure determination shows formation of a laminated sheet structure by hydrogen bonding between amide NH groups and triflate anions of the type NH-OSO-HN. PMID- 12639131 TI - Small organoselenium molecules. 1. Dimethyl selenoxide: structure, complexation, and gas-phase transformation. AB - For the first time the structural characterization of dimethyl selenoxide coordinated to metal complexes has been performed confirming the Me(2)SeO arrangement assigned by spectroscopic techniques for the molecule in solution and solid state. The structure of Me(2)SeO is trigonal pyramidal with Se-O and Se-C bond lengths of 1.70 and 1.92 A, respectively, and sigma(X-Se-Y) = 301 degrees. As a ligand, dimethyl selenoxide was found to bind to the rhodium centers of various Lewis acidity strengths by using only its oxo functionality in both terminal and bridging fashions. This O-directed coordination preference contrasts with an ambidentate (-S and -O) binding character revealed by dimethyl sulfoxide upon formation of analogous donor-acceptor complexes. The study of dimethyl selenoxide in the gas phase at 135-140 degrees C resulted in a thermal degradation of this molecule. The major decomposition product has been entrapped by a metal complex and identified as dimethyl selenide. The isolation of the coordinated Me(2)Se fragment clearly demonstrates that the Me(2)SeO molecule is less thermally stable than Me(2)SO, which under similar reaction conditions shows no sign of decomposition at temperatures up to 160 degrees C. PMID- 12639132 TI - The crystal structure and magnetic properties of a new ferrimagnetic semiconductor: Ca21Mn4Sb18. AB - Single crystals of the new transition metal Zintl phase, Ca(21)Mn(4)Sb(18), were prepared by high temperature melt synthesis. The crystal structure was determined by single crystal X-ray diffraction to be monoclinic in the space group C2/c. Crystal information was obtained at 90 K, and unit cell parameters were determined (a = 17.100(2) A, b = 17.073(2) A, c = 16.857(2) A, beta = 92.999(2) degrees, Z = 2, R1 = 0.0540, wR2 = 0.1437). The structure can be described as containing 4 discreet units per formula unit: 1 linear [Mn(4)Sb(10)](22-) anion, 2 dumbbell-shaped [Sb(2)](4-) anions, 4 individual Sb(3-) anions, and 21 Ca(2+) cations. The [Mn(4)Sb(10)](22-) anion contains four edge-shared MnSb(4) tetrahedra with distances between Mn ions of 3.388(4) A, 2.782(4) A, and 2.760(4) A. Electron counting suggests that the Mn are 2+. Temperature dependent magnetization shows a ferromagnetic-like transition temperature at approximately 52 K which is suppressed with increasing magnetic field. The paramagnetic regime is best fit to a ferrimagnetic model, providing a total effective moment of 4.04(2) mu(B), significantly less than that expected for 4 Mn(2+) ions (11.8 mu(B)). Temperature dependent resistivity shows that this compound is a semiconductor with an activation energy of 0.159(2) eV (100-300 K). PMID- 12639133 TI - Structure and bonding in solution of dioxouranium(VI) oxalate complexes: isomers and intramolecular ligand exchange. AB - Structural isomers of [UO(2)(oxalate)(3)](4-), [UO(2)(oxalate)F(3)](3-), [UO(2)(oxalate)(2)F](3-), and [UO(2)(oxalate)(2)(H(2)O)](2-) have been studied by using EXAFS and quantum chemical ab initio methods. Theoretical structures and their relative energies were determined in the gas phase and in water using the CPCM model. The most stable isomers according to the quantum chemical calculations have geometries consistent with the EXAFS data, and the difference between measured and calculated bond distances is generally less than 0.05 A. The complex [UO(2)(oxalate)(3)](4-) contains two oxalate ligands forming five membered chelate rings, while the third is bonded end-on to a single carboxylate oxygen. The most stable isomer of the other two complexes also contains the same type of chelate-bonded oxalate ligands. The activation energy for ring opening in [UO(2)(oxalate)F(3)](3-), deltaU++ = 63 kJ/mol, is in fair agreement with the experimental activation enthalpy, deltaH++ = 45 +/- 5 kJ/mol, for different [UO(2)(picolinate)F(3)](2-) complexes, indicating similar ring-opening mechanisms. No direct experimental information is available on intramolecular exchange in [UO(3)(oxalate)(3)](4-). The theoretical results indicate that it takes place via the tris-chelated intermediate with an activation energy of deltaU++ = 38 kJ/mol; the other pathways involve multiple steps and have much higher activation energies. The geometries and energies of dioxouranium(VI) complexes in the gas phase and solvent models differ slightly, with differences in bond distance and energy of typically less than 0.06 A and 10 kJ/mol, respectively. However, there might be a significant difference in the distance between uranium and the leaving/entering group in the transition state, resulting in a systematic error when the gas-phase geometry is used to estimate the activation energy in solution. This systematic error is about 10 kJ/mol and tends to cancel when comparing different pathways. PMID- 12639134 TI - Coordination properties of tris(2-carboxyethyl)phosphine, a newly introduced thiol reductant, and its oxide. AB - Acid-base properties and metal-binding abilities of tris(2-carboxyethyl)phosphine (TCEP), a newly introduced thiol group protectant, were studied in solution, using potentiometry, (1)H and (31)P NMR, and UV-vis spectroscopy, and also in the solid state by X-ray diffraction. Stability constants of complexes of the P-oxide of TCEP (TCEPO) were established by potentiometry. The list of metal ions studied included Ni(II), Cu(II), Zn(II), Cd(II), and Pb(II). Cu(II) catalyzed oxidation of TCEP to TCEPO. For all other systems ML complexes were found as major species at neutral pH with TCEP and TCEPO. Monoprotonated MHL species were also detected in weakly acidic conditions for all TCEP complexes and for the Pb(II) complex of TCEPO, while hydrolytic MH(-1)L complexes were found for TCEP at the weakly alkaline pH range. The NiL(4) complex was found to form at excess of TCEP. Overall, the complexes were found to be rather weak, with log beta(ML) values around 3-5 for TCEP and 1.5-2.5 for TCEPO. The phosphorus pK(a) value for TCEP, 7.68, suggests that it can be a good buffer for studies at physiological pH. PMID- 12639135 TI - Amine-templated linear vanadium sulfates with different chain structures. AB - Amine-templated vanadium sulfates of the formula [HN(CH(2))(6)NH][(V(IV)O)(2)(OH)(2)(SO(4))(2)].H(2)O, I, [H(3)N(CH(2))(2)NH(3)][V(III)(OH)(SO(4))(2)].H(2)O, II, and [H(2)N(CH(2))(4)NH(2)][(V(IV)O)(H(2)O)(SO(4))(2)], III, have been prepared under hydrothermal conditions. These vanadium sulfates add to the new emerging family of organically templated metal sulfates. Compound I has a linear chain structure consisting of V(2)O(8) square-pyramid dimers connected by corner-sharing SO(4) tetrahedra, creating four-membered rings along the chain. Both II and III possess simple linear chain topologies formed by VO(6) octahedra and SO(4) tetrahedra, with II having the tancoite chain structure. Compound I crystallizes in the triclinic space group P1 (No. 2) with a = 7.4852(4) A, b = 9.5373(5) A, c = 11.9177(6) A, alpha = 77.22 degrees, beta = 76.47(2) degrees, gamma = 80.86 degrees, Z = 2. Compound II: monoclinic, space group P2(1)/c (No. 14), a = 6.942(2) A, b = 10.317(3) A, c = 15.102(6) A, beta = 90.64(4) degrees, Z = 4. Compound III: triclinic, space group P1 (No. 2) with a = 6.2558(10) A, b = 7.0663(14) A, c = 15.592(4) A, alpha = 90.46(2) degrees, beta = 90.47(2) degrees, gamma = 115.68(2) degrees, Z = 2. Magnetic susceptibility measurements reveal weak antiferromagnetic interactions in I and III and ferromagnetic interactions in II. PMID- 12639136 TI - Molecular precursors for ferroelectric materials: synthesis and characterization of Bi2M2(mu-O)(sal)4(Hsal)4(OEt)2 and BiM4(mu-O)4(sal)4(Hsal)3(O(i)Pr)4 (sal = O2CC6H4O, Hsal = O2CC6H4OH) (M = Nb, Ta). AB - Reactions between triphenyl bismuth, salicylic acid, and niobium or tantalum ethoxide have been explored. Four new coordination complexes incorporating bismuth and the group 5 metals niobium or tantalum have been synthesized and characterized spectroscopically, by elemental analysis, and by single crystal X ray diffraction. The new complexes are Bi(2)M(2)(mu-O)(sal)(4)(Hsal)(4)(OEt)(2) (1a, M = Nb; 1b, M = Ta) and BiM(4)(mu-O)(4)(sal)(4)(Hsal)(3)(O(i)Pr)(4) (sal = O(2)CC(6)H(4)-2-O, Hsal = O(2)CC(6)H(4)-2-OH) (2a, M = Nb; 2b, M = Ta). Complexes 1a and 1b are isomorphous, as are 2a and 2b. The thermal and hydrolytic decomposition of 1a has been explored by DT/TGA and powder X-ray diffraction, while scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX) were used to characterize the morphology and composition of the oxides. The heterobimetallic molecules are completely converted to the amorphous bimetallic oxide by heating to 500 degrees C in air. Decomposition of 1a or 1b at 650 degrees C produces the metastable high temperature form of BiNbO(4) as the major crystalline oxide phase. Heating samples of 1a to 850 degrees C favors conversion of the materials to the low temperature phase as well as disproportionation into Bi(5)Nb(3)O(15) and Nb(2)O(5). Thermal decomposition of 1a and 1b produces porous oxides, while hydrolytic decomposition of the complexes has been shown to produce nanometer scale bimetallic oxide particles. The potential of the complexes to act as single-source precursors for ferroelectric materials is considered. PMID- 12639138 TI - Semiconducting perovskites (2-XC6H4C2H4NH3)2SnI4 (X = F, Cl, Br): steric interaction between the organic and inorganic layers. AB - Two new semiconducting hybrid perovskites based on 2-substituted phenethylammonium cations, (2-XC(6)H(4)C(2)H(4)NH(3))(2)SnI(4) (X = Br, Cl), are characterized and compared with the previously reported X = F compound, with a focus on the steric interaction between the organic and inorganic components. The crystal structure of (2-ClC(6)H(4)C(2)H(4)NH(3))(2)SnI(4) is solved in a disordered subcell [C2/m, a = 33.781(7) A, b = 6.178(1) A, c = 6.190(1) A, beta = 90.42(3)(o), and Z = 2]. The structure is similar to the known (2 FC(6)H(4)C(2)H(4)NH(3))(2)SnI(4) structure with regard to both the conformation of the organic cations and the bonding features of the inorganic sheet. The (2 BrC(6)H(4)C(2)H(4)NH(3))(2)SnI(4) system adopts a fully ordered monoclinic cell [P2(1)/c, a = 18.540(2) A, b = 8.3443(7) A, c = 8.7795(7) A, beta = 93.039(1)(o), and Z = 2]. The organic cation adopts the anti conformation, instead of the gauche conformation observed in the X = F and Cl compounds, apparently because of the need to accommodate the additional volume of the bromo group. The steric effect of the bromo group also impacts the perovskite sheet, causing notable distortions, such as a compressed Sn-I-Sn bond angle (148.7(o), as compared with the average values of 153.3 and 154.8(o) for the fluoro and chloro compounds, respectively). The optical absorption features a substantial blue shift (lowest exciton peak: 557 nm, 2.23 eV) relative to the spectra of the fluoro and chloro compounds (588 and 586 nm, respectively). Also presented are transport properties for thin-film field-effect transistors (TFTs) based on spin-coated films of the two hybrid semiconductors. PMID- 12639137 TI - Synthesis and structural characterization of a novel mixed-valent Cu(II)Cu(I)Cu(II) triangular metallomacrocycle using an imine-based rigid ligand. AB - A novel neutral mixed-valent Cu(I)Cu(II)(2) triangular metallomacrocycle [Cu(3)L(2)(HL)].3CH(3)OH.2H(2)O (1) was assembled by reaction of the tetradentate ligand bis(N-salicylidene-4,4'-diphenylamine), H(2)L, with a copper(II) salt. ESI MS show peaks only corresponding to the triangular structural species, indicating the high stability of the trimer structure in solution. Magnetic study confirms that there are two Cu(II) ions and one Cu(I) ion in a discrete triangular molecule. The crystal structure of 1 reveals that the triangle is formed by three deprotonated ligands and three copper ions with a Cu-Cu separation of ca. 11.8 A. Each copper atom is coordinated by two oxygen atoms and two nitrogen atoms from two different bis-bidentate ligands in a heavily distorted tetrahedral geometry, while each ligand is bound to two metal ions in a bis-bidentate coordination mode and links the metal centers overlapping in an unprogressive manner. Strong intramolecular pi.pi interactions between the ligands are found to stabilize the constraint conformation of the triangle. Electrochemical study reveals that the mixed-valent Cu(I)Cu(II)(2) complex is the most stable state in solution condition, and the electrochemical communication between the copper ions might be explained on the basis of the through-bond interaction. UV-vis-NIR spectral measurement demonstrates the Robin-Day class II behavior of the mixed-valence compound with a weak copper-copper interaction. PMID- 12639139 TI - Synthesis, structure, and spectroscopic, photochemical, redox, and catalytic properties of ruthenium(II) isomeric complexes containing dimethyl sulfoxide, chloro, and the dinucleating bis(2-pyridyl)pyrazole ligands. AB - Two isomeric Ru(II) complexes containing the dinucleating Hbpp (3,5-bis(2 pyridyl)pyrazole) ligand together with Cl and dmso ligands have been prepared and their structural, spectroscopic, electrochemical, photochemical, and catalytic properties studied. The crystal structures of trans,cis [Ru(II)Cl(2)(Hbpp)(dmso)(2)], 2a, and cis(out),cis-[Ru(II)Cl(2)(Hbpp)(dmso)(2)], 2b, have been solved by means of single-crystal X-ray diffraction analysis showing a distorted octahedral geometry for the metal center where the dmso ligands coordinate through their S atom. 1D and 2D NMR spectroscopy corroborates a similar structure in solution for both isomers. Exposure of either 2a or 2b in acetonitrile solution under UV light produces a substitution of one dmso ligand by a solvent molecule generating the same product namely, cis(out) [Ru(II)Cl(2)(Hbpp)(MeCN)(dmso)], 4. While the 1 e(-) oxidation of 2b or cis(out),cis-[Ru(II)Cl(2)(bpp)(dmso)(2)](+), 3b, generates a stable product, the same process for 2a or trans,cis-[Ru(II)Cl(2)(bpp)(dmso)(2)](+), 3a, produces the interesting linkage isomerization phenomenon where the dmso ligand switches its bond from Ru-S to Ru-O (K(III)(O)(-->)(S) = 0.25 +/- 0.025, k(III)(O)(-->)(S) = 0.017 s(-1), and k(III)(S)(-->)(O) = 0.065 s(-1); K(II)(O)(-->)(S) = 6.45 x 10(9), k(II)(O)(-->)(S) = 0.132 s(-1), k(II)(S)(-->)(O) = 2.1 x 10(-11) s(-1)). Finally complex 3a presents a relatively high activity as hydrogen transfer catalyst, with regard to its ability to transform acetophenone into 2-phenylethyl alcohol using 2-propanol as the source of hydrogen atoms. PMID- 12639141 TI - Complexing ability of the versatile, redox-active, 3-[3 (diphenylphosphino)propylthio]-3',4,4'-trimethyl-tetrathiafulvalene ligand. AB - The synthesis of a ligand containing as an electroactive core a tetrathiafulvalene moiety, 3-[3-(diphenylphosphino)propylthio]-3',4,4'-trimethyl tetrathiafulvalene, is reported. Its versatile ability to act as a bidentate or a monodentate ligand, as demonstrated by the metal carbonyl complexes obtained, is described. The novel cis-Mo(CO)(4)(P-TTF)(2) 4 and cis-W(CO)(4)(P,S-TTF) 6 complexes have been characterized by X-ray diffraction analyses and cyclic voltammetry measurements. Within complex 4, no significant influence of the two electroactive ligands on the molybdenum center was detected, whereas, in complex 6, a weak influence of the TTF redox-active core can be observed on the redox behavior of the metal center. PMID- 12639142 TI - Luminescent nido-carborane-diphosphine anions [(PR2)2C2B9H10](-) (R = Ph, (i)Pr). Modification of their luminescence properties upon formation of three-coordinate gold(I) complexes. AB - The free nido-diphosphine anions [(PR(2))(2)C(2)B(9)H(10)](-) (R = Ph, (i)Pr) show luminescence properties whereas the closo-diphosphines [(PR(2))(2)C(2)B(10)H(10)] do not. Four families of three-coordinate complexes of stoichiometry [Au[(PR(2))(2)C(2)B(10)H(10)]L]OTf (L = tertiary phosphine) and [Au[(PR(2))(2)C(2)B(9)H(10)]L] have been studied in order to analyze the influence of the closo- or nido-nature of the diphosphine, the monophosphine coordinated to gold and the substituent at the diphosphine on the luminescence of the complexes. Only the nido-derivatives show luminescence. The maxima of the emissions are shifted to lower energies than those of the corresponding free nido diphosphines. When the substituent at the diphosphine is phenyl, a new emission appears, which has been assigned as arising from a metal to ligand charge transfer [Au-->pi(L)] excited state. PMID- 12639140 TI - Cu(II) complexes with heterocyclic substituted thiosemicarbazones: the case of 5 formyluracil. Synthesis, characterization, x-ray structures, DNA interaction studies, and biological activity. AB - Two new 5-formyluracil thiosemicarbazone (H(3)ut) derivatives, Me-H(3)ut (1) and Me(2)-H(3)ut (2), were synthesized by reacting thiosemicarbazides, mono- and dimethylated on the aminic nitrogen, with 5-formyluracil and were subsequently characterized. These ligands, treated with copper chloride and nitrate, afforded three complexes: [Cu(Me-H(3)ut)Cl(2)].H(2)O (3), [Cu(Me(2)-H(3)ut)Cl(2)].H(2)O (4), and [Cu(Me-H(3)ut)(NO(3))(OH(2))(2)]NO(3) (5). The crystal structures of these complexes have been determined by single-crystal X-ray diffraction. In 3 and 4, a similar pentacoordination is present; the copper atom is surrounded by the ligand SNO donor atoms and by two chloride ions. The structure of 5 consists of [Cu(Me-H(3)ut)(NO(3))(OH(2))(2)](+) cations and nitrate anions. The copper coordination (4 + 2) involves the SNO ligand atoms and a water oxygen in the basal plane; the apical positions are occupied by a second water oxygen and by an oxygen of a monodentate nitrate group. Two biochemical techniques, namely DNA titration in the UV-vis region and thermal denaturation, have been employed to probe the details of DNA binding of these compounds. Analysis of the results suggests that our compounds are able to interact with DNA by electrostatic and groove binding but not by intercalation. The compounds have been also tested in vitro on human leukemic cell line U937, but they are not able to inhibit significantly cell proliferation. PMID- 12639143 TI - Unprecedented chemical transformation of benzaldehyde semicarbazone mediated by osmium. AB - para-Nitrobenzaldehyde semicarbazone (O(2)N(para)-C(6)H(4)C(H)=N-NH-CO-NH(2)) undergoes unprecedented chemical transformation during its reaction with [Os(PPh(3))(2)(CO)(2)(HCOO)(2)] in different alcoholic (R'OH, R' = CH(2)CH(2)OCH(3), CH(2)CH(3), CH(2)CH(2)CH(3), and CH(2)CH(2)CH(2)CH(3)) solvents whereby the NH(2) group of the semicarbazone ligand is displaced by a OR' group provided by the solvents. The transformed semicarbazone ligand binds to osmium as a bidentate N,O-donor forming five-membered chelate ring to afford complexes of type [Os(PPh(3))(2)(CO)(H)(L-OR')], where L-OR' refers to the transformed semicarbazone ligand. Structure of the [Os(PPh(3))(2)(CO)(H)(L OCH(2)CH(2)OCH(3))] complex has been determined by X-ray crystallography. All the [Os(PPh(3))(2)(CO)(H)(L-OR')] complexes are diamagnetic and show characteristic (1)H NMR signals. They also show intense absorptions in the visible and ultraviolet region. Cyclic voltammetry on the complexes shows an irreversible oxidative response within 0.69-0.88 V versus SCE. PMID- 12639145 TI - Syntheses and X-ray structures of monocyclic, bicyclic, and spirocyclic gallium and indium boraamidinates. AB - The reactions of [Li(2)[PhB(N(t)Bu)(2)]](2) with GaCl(3) in various stoichiometries yield [Li(thf)(4)][PhB(mu-N(t)Bu)(2)GaCl(2) x GaCl(3)] (1), [PhB(mu-N(t)Bu)(2)GaCl](2) (2), and [mu-Li(OEt(2))[PhB(N(t)Bu)(2)]Ga] (3a), a series of complexes in which the three chloride ligands are successively replaced by the dianion [PhB(N(t)Bu)(2)](2-). The X-ray structures of 1, 2, and 3a show that the boraamidinate ligand adopts an N,N'-chelating mode. In the ion-separated complex 1, one of the nitrogen atoms is coordinated to a GaCl(3) molecule. The related indium complexes [mu-LiCl(thf)(2)][PhB(mu-N(t)Bu)(2)InCl](2) (4) and [mu Li(OEt(2))[PhB(mu-N(t)Bu)(2)]In] (3b) were obtained in a similar manner. Complex 4 is the indium analogue of 2 with the incorporation of a bissolvated LiCl molecule. In 3a and 3b the spirocyclic [[PhB(mu-N(t)Bu)(2)](2)M](-) (M = Ga, In) anions are N,N'-chelated to the [Li(OEt(2))](+) counterion. Prolonged reactions result in the formation of [PhB(mu-N(t)Bu)(2)GaCl][(t)BuN(H)GaCl(2)] (5) and [[PhB(mu-N(t)Bu)(2)InCl][(t)BuN(H)InCl(2)][mu-LiCl(OEt(2))(2)]] (6), respectively. The X-ray structures of 5 and 6 reveal bicyclic structures which formally involve the entrapment of the monomers (t)BuN(H)MCl(2) by a four membered BN(2)M ring (M = Ga, In). The synthesis and X-ray structure of Cl(2)Ga[mu-N(H)(t)Bu](2)GaCl(2) are also reported. PMID- 12639144 TI - Porphyrins linked directly to the 5,5' positions of 2,2'-bipyridine: a new supramolecular building block and switch. AB - The synthesis and coordination chemistry of two porphyrin dimers linked either at the 5,5' or the 4,4' positions of 2,2'-bipyridine are described. These compounds, which may serve a molecular tectons for the constructions of a variety of supramolecular arrays of diverse function, reveal that the ground- and excited state electronic communication between the chromophores is only moderately affected by the complexation state of the bipyridyl moiety. The nature of the metal ion chelated by the bipyridine only slightly perturbs the ground-state spectra, and differences observed in the excited state are largely ascribed to the heavy atom effect. This work also shows that conformational changes in structural subunits, in this case induced by bipyridyl complexation of various metal ions, do not necessarily require reorganization of supramolecular systems. PMID- 12639146 TI - Remarkable dynamical opening and closing of platinum and palladium pentaruthenium carbido carbonyl cluster complexes. AB - The reaction of Ru(5)(CO)(15)(mu(5)-C), 1, with Pt(PBu(t)(3))(2) at room temperature yielded the mixed-metal cluster complex PtRu(5)(CO)(15)(PBu(t)(3))(C), 2, in 52% yield. Compound 2 consists of a mixture of two interconverting isomers in solution. One isomer, 2A, can be isolated by crystallization from benzene/octane solvent. The second isomer, 2B, can be isolated by crystallization from diethyl ether. Both were characterized crystallographically. Isomer 2A consists of a square pyramidal cluster of five ruthenium atoms with a phosphine-substituted platinum atom spanning the square base. Isomer 2B consists of a square pyramidal cluster of five ruthenium atoms with a phosphine-substituted platinum atom on an edge on the square base. The two isomers interconvert rapidly on the NMR time scale at 40 degrees C, deltaG(313)++ = 11.4(8) kcal mol(-1), deltaH++ = 8.8(5) kcal mol(-1), deltaS++ = -8.4(9) cal mol(-1) K(-1). The reaction of Pd(PBu(t)(3))(2) with compound 1 yielded two new cluster complexes: PdRu(5)(CO)(15)(PBu(t)(3))(mu(6)-C), 3, in 50% yield and Pd(2)Ru(5)(CO)(15)(PBu(t)(3))(2)(mu(6)-C), 4, in 6% yield. The yield of 4 was increased to 47% when an excess of Pd(PBu(t)(3))(2) was used. In the solid state compound 3 is structurally analogous to 2A, but in solution it also exists as a mixture of interconverting isomers; deltaG(298)++ = 10.6(6) kcal mol(-1), deltaH++ = 9.7(3) kcal mol(-1), and deltaS++ = -3(1) cal mol(-1) K(-1) for 3. Compound 4 contains an octahedral cluster consisting of one palladium atom and five ruthenium atoms with an interstitial carbido ligand in the center of the octahedron, but it also has one additional Pd(PBu(t)(3)) grouping that is capping a triangular face of the ruthenium cluster. The Pd(PBu(t)(3)) groups in 4 also undergo dynamical interchange that is rapid on the NMR time scale at 25 degrees C; deltaG(298)++ = 11(1) kcal mol(-1), deltaH++ = 10.2(4) kcal mol(-1), and deltaS++ = -3(2) cal mol(-1) K(-1) for 4. PMID- 12639147 TI - Solid-State and solution studies of [Ln(n)(SiW11O39)] polyoxoanions: an example of building block condensation dependent on the nature of the rare earth. AB - The reactivity of the [alpha-SiW(11)O(39)](8-) monovacant polyoxometalate with lanthanide has been investigated for four different trivalent rare-earth cations (Ln = Nd(III), Eu(III), Gd(III), Yb(III)). The crystal structures of KCs(4)[Yb(alpha-SiW(11)O(39))(H(2)O)(2)] x 24H(2)O (1), K(0.5)Nd(0.5)[Nd(2)(alpha SiW(11)O(39))(H(2)O)(11)] x 17H(2)O (2a), and Na(0.5)Cs(4.5)[Eu(alpha SiW(11)O(39))(H(2)O)(2)] x 23H(2)O (3a) are reported. The solid-state structure of compound 1 consists of linear wires built up of [alpha-SiW(11)O(39)](8-) anions connected by Yb(3+) cations, while the linkage of the building blocks by Eu(3+) centers in 3a leads to the formation of zigzag chains. In 2a, dimeric [Nd(2)(alpha-SiW(11)O(39))(2)(H(2)O)(8)](10-) entities are linked by four Nd(3+) cations. The resulting chains are connected by lanthanide ions, leading to a bidimensional arrangement. Thus, the dimensionality, the organization of the polyoxometalate building units, and the Ln/[alpha-SiW(11)O(39)](8-) ratio in the solid state can be tuned by choosing the appropriate lanthanide. The luminescent properties of compound 3a have been studied, showing that, in solution, the polymer decomposes to give the monomeric complex [Eu(alpha SiW(11)O(39))(H(2)O)(4)](5-). The lability of the four exogenous ligands connected to the rare earth must allow the functionalization of this lanthanide polyanion. PMID- 12639148 TI - Versatile ligand behavior of hydrotris(4-ethyl-3-methyl-5-thioxo-1,2,4 triazolyl)borate. Syntheses and crystal structures of Cu(I) and Bi(III) complexes. AB - Preparations of copper(I) and bismuth(III) complexes of hydrotris(4-ethyl-3 methyl-5-thioxo-1,2,4-triazolyl)borate (Tr(Et,Me)) are described. These complexes have been characterized by means of spectroscopy and microanalysis. Molecular structures of [Cu(Tr(Et,Me))](2) x 2.5CH(3)CN x 0.5H(2)O (3a) and [Bi(Tr(Et,Me))(2)]NO(3) x 2CHCl(3) (4a) have been determined by single-crystal X ray diffraction. In the centrosymmetric dimeric copper(I) complex, Tr(Et,Me) acts in the k(3)S,S',H:kS' ' coordination mode. The metal is found in a distorted trigonal geometry as the ligand exhibits an "S(3)-inverted" conformation at the boron center so that a weak [B-H.Cu] agostic interaction renders the overall coordination of the (3 + 1) type. On the other hand, in the bismuth complex, Tr(Et,Me) presents the k(3)S,S',S' ' coordination mode and the "S(3)-normal" conformation. The metal is found in a regular octahedral geometry bound by six thioxo groups of two ligands. Species distributions in solution have been studied using electrospray ionization mass spectrometry upon dissolution of 3a and 4a crystals in acetonitrile. Monomeric and polynuclear copper(I) complexes with different M:L ratios are present in solution, while for 4a only the monomeric species is present. PMID- 12639149 TI - 1H,1H,2H,2H-perfluoroalkyl-functionalization of Ni(II), Pd(II), and Pt(II) mono- and diphosphine complexes: minimizing the electronic consequences for the metal center. AB - A series of fluorous derivatives of group 10 complexes MCl(2)(dppe) and [M(dppe)(2)](BF(4))(2) (M = Ni, Pd or Pt; dppe = 1,2 bis(diphenylphosphino)ethane) and cis-PtCl(2)(PPh(3))(2) was synthesized. The influence of para-(1H,1H,2H,2H-perfluoroalkyl)dimethylsilyl-functionalization of the phosphine phenyl groups of these complexes, as studied by NMR spectroscopy, cyclovoltammetry (CV), XPS analyses, as well as DFT calculations, points to a weak steric and no significant inductive electronic effect. The steric effect is most pronounced for M = Ni and leads in the case of NiCl(2)(1c) (3c) and [Ni(1c)(2)](BF(4))(2) (7c) (1c = [CH(2)P[C(6)H(4)(SiMe(2)CH(2)CH(2)C(6)F(13)) 4](2)](2)) to a tetrahedral distortion from the expected square planar geometry. The solubility behavior of NiCl(2)[CH(2)P[C(6)H(4)(SiMe(3 b)(CH(2)CH(2)C(x)F(2x+1)b)-4](2)](2) (3: b = 1-3; x = 6, 8) in THF, toluene, and c-C(6)F(11)CF(3) was found to follow the same trends as those observed for the free fluorous ligands 1. A similar correlation between the partition coefficient (P) of complexes 3 and free 1 was observed in fluorous biphasic solvent systems, with a maximum value obtained for 3f (b = 3, x = 6, P = 23 in favor of the fluorous phase). PMID- 12639150 TI - Understanding structure does not always explain reactivity: a phosphinoamide anion reacts as an iminophosphide anion. AB - The aminophosphine C(6)H(4)(o-CN)NHPPh(2), 1, containing an electron-withdrawing nitrile group, was prepared from the lithiation of 2-aminobenzonitrile followed by addition of Ph(2)PCl. Lithiation of 1 using (n)BuLi affords the anion [Ph(2)PN(Li)C(6)H(4)(o-CN)(thf)](2), 2. Compound 2 reacts with Ph(2)PCl or MeI to afford C(6)H(4)(o-CN)N=PPh(2)-PPh(2), 3, and [C(6)H(4)(o CN)N=PPh(2)(CH(3))(LiI)(C(2)H(5)O)](2), 4, respectively. In these products new P P and P-C bonds have been formed rather than N-P and N-C bonds. The structures of 1-4 have been determined by single-crystal X-ray diffraction analysis, and the synthetic results are discussed in terms of the structural data and NMR spectroscopic studies. PMID- 12639151 TI - Synthesis and characterization of trifluoromethyl fluoroformyl peroxycarbonate, CF3OC(O)OOC(O)F. AB - The synthesis of CF(3)OC(O)OOC(O)F is accomplished by the photolysis of a mixture of (CF(3)CO)(2)O, FC(O)C(O)F, CO, and O(2) at -15 degrees C using a low-pressure mercury lamp. The new peroxide is obtained in pure form in low yield after repeated trap-to-trap condensation and is characterized by NMR, IR, Raman, and UV spectroscopy. Geometrical parameters were studied by ab initio methods [B3LYP/6 311+G(d)]. At room temperature, CF(3)OC(O)OOC(O)F is stable for many days in the liquid or gaseous state. The melting point is -87 degrees C, and the boiling point is extrapolated to 45 degrees C from the vapor pressure curve log p = 8.384 - 1715/T (p/mbar, T/K). A possible mechanism for the formation of CF(3)OC(O)OOC(O)F is discussed, and its properties are compared with those of related compounds. PMID- 12639152 TI - [Cu(H4C3N2S)Cl2](n), an unprecedented diazole-bridged one-dimensional copper halide: synthesis, structure, and magnetic properties. AB - The simultaneous desulfurization of 2-mercapto-5-methyl-1,3,4-thiadiazole with CuCl(2) x 2H(2)O via mutual diffusion in solvents results in the isolation of air stable dark-green crystals of [Cu(H(4)C(3)N(2)S)Cl(2)](n) (approximately 65% yield). The structure is characterized by a unique one-dimensional copper chain bridged by diazine N-N single bonds rather than halogens, in sharp contrast with the halide bridging mode in conventional copper halide coordination polymers. Each Cu(II) ion shows a square planar coordination geometry featuring a strong Jahn-Teller distortion, as also supported by EPR data. The phase follows a Curie Weiss paramagnetic behavior over 6-300 K. However, the intrachain antiferromagnetic interaction is evident (-2J = 21.1 cm(-1)). Such magnetic coupling is related to the interplay between the Cu(II)-d(x2-y2) and diazine N-N p-orbitals. PMID- 12639153 TI - Three- and four-coordinate gold(I) complexes of 3,6 bis(diphenylphosphino)pyridazine: monomers, polymers, and a metallocryptand cage. AB - The slightly yellow polymeric complexes [Au(2)Cl(2)(P(2)pz)(3)](n), 1 x 6CHCl(3), (P(2)pz is 3,6-bis(diphenylphosphino)pyridazine) and [[Au(2)(P(2)pz)(3)](PF(6))(2)](n), 2, are prepared by the stoichiometric reaction of AuCl(tht) (tht is tetrahydrothiophene) and P(2)pz in either dichloromethane or dichloromethane/methanol, respectively. Addition of 2 equiv of AuCl(tht) to a dichloromethane solution of 1 equiv of P(2)pz generates the simple (AuCl)(2)(P(2)pz) compound, 3. Compound 3 contains nearly linear P-Au-Cl units with intermolecular Au.Au separations of 3.570 A. Au(2)I(2)(P(2)pz)(3), 4, is prepared by reacting excess NaI with 2 in a dichloromethane/methanol mixture. Characterization of 1, 2, and 4 by X-ray crystallography confirms the 2:3 gold/ligand ratio of all three complexes. The coordination polymer 1 maintains a high degree of solvation in the solid-state with three chloroform adducts hydrogen-bonded to the chloride ligand on each gold atom. These chloroform molecules are sandwiched between the two-dimensional polymeric sheets of 1. The crystal structure of 4 reveals an empty, iodide-capped metallocryptand cage with the tetrahedrally distorted gold atoms and the nitrogen atoms on the pyridazine rings directed away from the center of the cavity. No metal ion encapsulation was observed for complex 4. Complex 2 forms one-dimensional arrays of [Au(2)(P(2)pz)(2)](2+) metallomacrocycles connected to each other by a third P(2)pz ligand. The electronic absorption spectra (CH(2)Cl(2)) of 1-4 show broad, nearly featureless absorption bands that tail into the visible with pi-pi bands at 296 nm and discernible shoulders at 314 nm for 2 and 334 nm for 3. Excitation into the low energy band of 2 produces only a modest emission in solution at 540 nm (lambda(ex) 468 nm) and 493 nm (lambda(ex) 403 nm). Under identical conditions, the P(2)pz ligand also emits at 540 and 493 nm. PMID- 12639154 TI - Homoleptic group 12 metal bis(mercaptoimidazolyl)borate complexes M(Bm(R))2 (M = Zn, Cd, Hg). AB - The sodium salt of the bis(2-mercapto-1-methylimidazolyl)borate anion [Bm(Me)](-) and those of the new bis(2-mercapto-1-alkylimidazolyl)borates [Bm(R)](-) (R = Bz, Bu(t), p-Tol) have been readily obtained from NaBH(4) and the appropriate 2 mercapto-1-alkylimidazoles. To contrast the binding preferences of the group 12 metals in a sulfur-rich environment, the four complete series of homoleptic complexes M[Bm(R)](2) (M = Zn, Cd, Hg), including the first bis(mercaptoimidazolyl)borate derivatives of cadmium and mercury, have been prepared. X-ray diffraction studies of Cd[Bm(Me)](2) and M[Bm(tBu)](2) (M = Zn, Cd, Hg) show the presence of distorted tetrahedral [MS(4)] central cores supplemented by two weak vicinal M.H-B bonds, interactions which appear to be a common feature in the coordination chemistry of Bm(R) ligands. In the case of zinc, it has been found that only in the presence of bulky ligands, as in Zn[Bm(tBu)](2), may an unexpected expansion in the coordination number from four to six be induced. This observation suggests the viability of octahedral intermediates in the processes whereby certain zinc enzymes transfer or exchange metal ions. PMID- 12639155 TI - Mixed dinitrogen-organocyanamide complexes of molybdenum(0) and their protic conversion into hydrazide and amidoazavinylidene derivatives. AB - Organocyanamides, Ntbd1;CNR(2) (R = Me or Et), react with trans [Mo(N(2))(2)(dppe)(2)] (1, dppe = Ph(2)PCH(2)CH(2)PPh(2)), in THF, to give the first mixed molybdenum dinitrogen-cyanamide complexes trans [Mo(N(2))(NCNR(2))(dppe)(2)] (R = Me 2a or Et 2b) which are selectively protonated at N(2) by HBF(4) to yield the hydrazide(2-) complexes trans [Mo(NNH(2))(NCNR(2))(dppe)(2)][BF(4)](2) (R = Me, 3a, or Et, 3b). On treatment with Ag[BF(4)], oxidation and metal fluorination occur, and the ligating cyanamide undergoes an unprecedented beta-protonation at the unsaturated C atom to form trans-[MoF(NCHNR(2))(dppe)(2)][BF(4)](2) (R = Me, 4a, or Et, 4b) compounds which present the novel amidoazavinylidene (or amidomethyleneamide) ligands. Complexes 4 are also formed from the corresponding compounds 3, with liberation of ammonia and hydrazine. The crystal structure of 2b was determined by single-crystal X-ray diffraction analysis which indicates that the N atom of the amide group has a trigonal planar geometry. PMID- 12639156 TI - Structural diversity and polytypism of lead phenylphosphonates: BING-6 and BING 9. AB - We report the solvothermal synthesis and characterization of a series of layered lead phenylphosphonates. The crystals were suitable for single crystal X-ray diffraction data, and the two new structures we denote BING-6 [SUNY at Binghamton, Structure No. 6, Pb(PO(3)C(6)H(5)).0.25C(5)H(5)N, triclinic space group Ponemacr;, Z = 2, a = 7.0770(4) A, b = 9.3113(6) A, c = 14.6785(9) A, alpha = 80.456(1) degrees, beta = 78.023(1) degrees, gamma = 73.265(1)(o)] and BING-9 [Pb(PO(3)HC(6)H(5))(PO(3)HC(6)H(4)CH(3)), monoclinic space group C2/c, Z = 4, a = 32.663(8) A, b = 5.6220(13) A, c = 8.3307(19) A, beta = 101.419(4)(o)]. The third structure, a polytype of BING-9, was previously known only from powder X-ray diffraction methods and is denoted 3 [Pb(PO(3)HC(6)H(5))(2), monoclinic space group C2/c, Z = 4, a = 31.681(6) A, b = 5.5639(11) A, c = 8.2515(16) A, beta = 101.814(4)(o)]. All three structures possess Pb(II) and P centers connected by doubly and triply bridging oxygens. The phenyl groups cap and separate the charge neutral layers. The phosphonates of BING-6 are nonprotonated, and the structure therefore has a Pb/P ratio of 1:1. Neutral, partially disordered pyridine solvent molecules also reside in the interlamellar space, increasing the layer to layer distance. BING-9 and 3 are polytypes and contain singly protonated phosphonates, for a Pb/P ratio of 1:2. Further characterization methods are discussed, including powder X-ray diffraction, in-situ variable temperature powder X-ray diffraction, thermogravimetric analysis, and scanning electron microscopy. Related work in the Ge, Sn, and Mn systems is also discussed. These low dimensional materials may be useful intercalation compounds for ion-exchange or sensor applications. PMID- 12639157 TI - Economic impact of ramipril on hospitalization of high-risk cardiovascular patients. AB - OBJECTIVE: To estimate differences in direct costs attributable to avoided hospitalizations and procedures during the years of the HOPE (Heart Outcomes Prevention Evaluation) study after the cost of treatment with ramipril or alternative angiotensin-converting enzyme inhibitor therapy was taken into account. METHODS: A decision analytical model was developed to estimate the economic impact of reductions in hospitalizations and/or procedures both at annual increments and over the first 4 years of the HOPE study. The analysis compared the number of cardiovascular events per endpoint per year in the intervention and placebo group with hospitalization and procedural costs. Cost data were derived from the literature and inflated to the appropriate index year using the consumer price index. RESULTS: For approximately 9000 patients studied, the gross estimated savings in direct costs for 297 events avoided were more than $5 million over 4 years. After the cost of treatment was deducted for both groups, the net estimated savings were $871 000 over 4 years. CONCLUSIONS: The results demonstrate that the use of ramipril provides cost-effective treatment for high-risk cardiovascular patients with an ejection fraction >40%. PMID- 12639159 TI - Ipratropium treatment of acute airways disease. AB - OBJECTIVE: To examine the use of ipratropium bromide in adults with acute exacerbation of either asthma or chronic obstructive pulmonary disease (COPD) during admission and at discharge from 3 teaching hospitals. METHODS: An extensive range of clinical and demographic data was retrospectively extracted from the medical records of consecutive patients aged >/=12 years admitted to the medical wards of the hospitals during 1999-2001 with acute exacerbations of asthma or COPD, either as a primary diagnosis or as a major comorbidity. RESULTS: Data were gathered for 302 patients (97 with asthma, 205 with COPD). Almost 90% of all patients received ipratropium bromide during their hospital admission. The indication for using ipratropium bromide during hospitalization was considered appropriate in 84% of the asthma patients and 68% of the COPD patients. Over 20% of the patients with asthma had been using ipratropium bromide prior to the hospital admission, and almost one-third of the patients with asthma were discharged with ipratropium bromide, even though its role in chronic asthma is limited. More than 90% of the patients received nebulized drug therapy during their hospital stay, with 41% being prescribed nebulized therapy at discharge. This was often considered inappropriate, given that >50% of all patients had been using inhaler devices and nebulizers concurrently while hospitalized. CONCLUSIONS: In the majority of cases, ipratropium bromide had been used during hospitalization in accordance with guidelines for the management of acute exacerbations of asthma or COPD. However, there was also evidence of potentially inappropriate prolonged use of the agent in the community setting, particularly for asthma. Also of concern was the relatively high use of nebulized drug therapy when delivery via other means was appropriate. PMID- 12639158 TI - Economic impact of pharmacists' interventions with nonsteroidal antiinflammatory drugs. AB - OBJECTIVE: To estimate the economic impact of community pharmacists' interventions following the detection of problems related to nonsteroidal antiinflammatory drugs (NSAIDs), whether in a prescription or self-medication format. The evaluation focused on the gastroduodenal adverse events that could be avoided and the subsequent savings of healthcare resources spent on treating these adverse effects. METHODS: A previous study conducted during a 12-week period in 924 French community pharmacies provided the number of interventions for drug-related problems concerning NSAIDs. A simulation model was constructed to compare 2 strategies: a systematic pharmacist's intervention and the absence of intervention. The base-case patient was assumed to have been taking an NSAID for 3 months. The model's inputs were extracted from medical literature and from an institutional medical database. RESULTS: In this study, 608 interventions were the results of NSAID-related problems. All of these interventions reduced the risk of gastrointestinal adverse events and avoided a total cost of 37 300. CONCLUSIONS: This model indicates that the dispensing of NSAIDs by pharmacists and related pharmaceutical care activities have a positive impact by reducing the number of gastrointestinal complications. The model quantifies the costs thus avoided. It also underlines the necessity of effective collaboration between the prescriber and the pharmacist if optimal patient management is to be achieved. PMID- 12639160 TI - Hemodynamic and electrocardiographic effects of short-term Ginkgo biloba. AB - OBJECTIVE: To evaluate the immediate and short-term hemodynamic and electrocardiographic effects of Ginkgo biloba (ginkgo). METHODS: Healthy volunteers were randomized to receive ginkgo 120 mg or placebo twice daily for 7 days in this prospective, double blind trial. After at least a 7-day washout period, subjects were crossed over to an additional 7 days of alternate therapy. Blood pressure, heart rate, and 12-lead electrocardiograms were evaluated immediately before (baseline), and at 1, 3, and 5 hours after observed ingestion of study drug on days 1 and 7 of therapy. Electrocardiographic parameters (P wave and QRS complex duration; PR, QT, and QTc intervals) were measured in lead II by a blinded investigator. RESULTS: Ginkgo had no effect on any of the evaluated electrocardiographic parameters at any time point on days 1 or 7. Additionally, no changes in heart rate or systolic and diastolic blood pressure were found between the groups at any time point on any evaluative day. CONCLUSIONS: Commonly used doses of Ginkgo biloba do not have any immediate or short-term effects on blood pressure, heart rate, or electrocardiographic variables in young, healthy volunteers. PMID- 12639161 TI - Branded versus generic clozapine for treatment of schizophrenia. AB - OBJECTIVE: To report clinical findings resulting from a switch from branded to generic clozapine. METHODS: Twenty patients diagnosed with schizophrenia were followed in this naturalistic outpatient study. The Positive and Negative Syndrome Scale (PANSS), Beck Anxiety Inventory (BAI), Abnormal Involuntary Movement Scale, and the Movement Disorder Assessment were used to assess differences in the clinical status of patients before and after switching from Clozaril to generic clozapine (Mylan Pharmaceuticals). Results were analyzed by means of the paired t-test and by calculation of the percent change in mean scores. A clinically significant change as measured by the PANSS was defined as a +/- 20% change in mean scores at final evaluation. The design was open-label and non-blinded. RESULTS: At the final evaluation, the t-test revealed no significant differences between branded and generic clozapine for the total PANSS, the positive symptom, negative symptom, and the general psychopathology subscales of the PANSS, and the BAI. There were no clinically significant changes for any measure. CONCLUSIONS: In this small group of patients with schizophrenia, no deterioration in clinical status in several domains was noted after changing from branded to generic clozapine. This finding is consistent with pharmacologic data suggesting bioequivalence of the 2 products. Results, however, must be interpreted cautiously due to the lack of optimal study controls and small sample size. PMID- 12639162 TI - Pharmacy practitioners' views on computerized documentation of drug-related problems. AB - OBJECTIVE: To evaluate the practice by community pharmacy practitioners of computerized documentation of drug-related problems (DRPs) and pharmacy interventions in nonprescription drug consumers. METHODS: A questionnaire was administered in December 1999 to pharmacy practitioners in 45 community pharmacies of different sizes and locations across Sweden to survey their attitudes and experiences after participation in a 10-week period of computerized DRP documentation. RESULTS: The participants (n = 376, response rate 84%) found the development of computerized documentation of DRPs and pharmacy interventions to be very important. The instrument was perceived as easy to learn and to work well in daily practice. The documentation made many practitioners more attentive to the drug-related needs of self-care consumers and changed their perception of good quality in self-care counseling. The weighted multiple linear regression analysis showed no correlation between the proportion of practitioners experiencing time constraints and the DRP documentation rate of their work site. However, the magnitude of interest in the documentation practice had a significant effect on the documentation rate, regardless of the extent of the time constraints experienced (p = 0.004). CONCLUSIONS: The positive findings of the evaluation speak in favor of an expanded implementation of computerized documentation of DRPs and pharmacy interventions. Commitment among participating pharmacy practitioners to the new practice is essential and appears to overcome possible time constraints. PMID- 12639163 TI - International survey of self-reported medicine use among adolescents. AB - OBJECTIVE: To examine gender, age, and country variations in adolescents' self reported medicine use. DESIGN: Cross-sectional school surveys of representative samples of 11- to 15-year-old girls and boys were used. The 1997/1998 Health Behaviour in School-aged Children study was referenced. A standardized questionnaire was completed during school hours. SETTING: Canada, US, Greenland, Israel, and 24 European countries. PARTICIPANTS: 123 227 participants equally distributed by gender and by 3 age groups (mean 11.7, 13.6, 15.6 y). MAIN OUTCOME MEASURES: Self-reported medicine use for headache, stomachache, difficulties in getting to sleep, and nervousness during the past month. RESULTS: The magnitude of the adolescents' medicine use for headache, stomachache, difficulties in getting to sleep, and nervousness varied substantially across countries. In each of the 28 countries, more girls than boys used medicine for pain. Use of medicine for headache increased by age; use of medicine for stomachache increased by age among girls, but decreased among boys; and use of medicine for difficulties in getting to sleep and nervousness decreased from the age of 11 to 15 years. There was an increase in the crude girl versus boy ratios for medicine use by age for all 4 symptoms. Multivariate logistic regression analyses, adjusting for age group and country, revealed the following odds ratios (95% CI) for girls' versus boys' medicine use: headache 1.56 (1.53 to 1.60), stomachache 2.16 (2.10 to 2.22), difficulties in getting to sleep 0.96 (0.91 to 1.00), and nervousness 1.04 (0.99 to 1.08). CONCLUSIONS: Substantial proportions of adolescents used medicine for common health problems. The prevalence of use differed between type of symptom for which the medicine was used, between countries, and between gender and age groups. We suggest that young people's medicine use should be addressed in public health policy. PMID- 12639164 TI - Warfarin and amoxicillin/clavulanate drug interaction. AB - OBJECTIVE: To report a case of warfarin-amoxicillin/clavulanate potassium (AM/CL) interaction resulting in an elevated international normalized ratio (INR) and hematuria. CASE SUMMARY: A 58-year-old Hawaiian/Asian/European woman developed an elevated INR and microscopic hematuria as a result of a drug-drug interaction between warfarin and AM/CL. DISCUSSION: Our report of an increased INR with bleeding complications as a result of an interaction between warfarin and AM/CL is consistent with those in the literature. Although the mechanism for this interaction is not fully known, it is suspected that a decrease in vitamin K producing gut flora with resulting vitamin K deficiency would be the most likely contributing factor. An objective causality assessment revealed that this adverse drug event as a result of the warfarin and AM/CL interaction was possible. CONCLUSIONS: An increased INR secondary to warfarin interactions with various antibacterial agents is a known phenomenon. An increased awareness of warfarin AM/CL interaction and appropriate monitoring are essential to control the INR levels and prevent bleeding complications. PMID- 12639165 TI - Ticlopidine-induced cholestatic hepatitis. AB - OBJECTIVE: To report 2 cases of ticlopidine-induced cholestatic hepatitis, investigate its mechanism, and compare the observed main characteristics with those of the published cases. CASE SUMMARIES: Two patients developed prolonged cholestatic hepatitis after receiving ticlopidine following percutaneous coronary angioplasty, with complete remission during the follow-up period. T-cell stimulation by therapeutic concentration of ticlopidine was demonstrated in vitro in the patients, but not in healthy controls. DISCUSSION: Cholestatic hepatitis is a rare complication of the antiplatelet agent ticlopidine; several cases have been reported but few in the English literature. Our patients developed jaundice following treatment with ticlopidine and showed the clinical and laboratory characteristics of cholestatic hepatitis, which resolved after discontinuation of the drug. Hepatitis may develop weeks after discontinuation of the drug and may run a prolonged course, but complete remission was observed in all reported cases. An objective causality assessment revealed that the adverse drug event was probably related to the use of ticlopidine. The mechanisms of this ticlopidine induced cholestasis are unclear. Immune mechanisms may be involved in the drug's hepatotoxicity, as suggested by the T-cell stimulation study reported here. CONCLUSIONS: Cholestatic hepatitis is a rare adverse effect of ticlopidine that may be immune mediated. Patients receiving the drug should be monitored with liver function tests along with complete blood cell counts. This complication will be observed even less often in the future as ticlopidine is being replaced by the newer antiplatelet agent clopidogrel. PMID- 12639166 TI - Intrapleural alteplase in a patient with complicated pleural effusion. AB - OBJECTIVE: To report the intrapleural use of alteplase in a patient diagnosed with complicated pleural effusion (CPE). CASE SUMMARY: A 62-year-old white woman admitted with respiratory distress and hypotension developed a right-sided multi loculated pleural effusion. Thoracentesis and chest tube drainage were not successful in resolving the effusion. In an attempt to increase the drainage of the pleural effusion, alteplase 16 mg was administered into the pleural cavity via the chest tube on 6 consecutive days. As a result, the volume drained from the patient's chest tube increased, there was improvement on the chest X-ray, and she did not require surgical intervention. DISCUSSION: While streptokinase and urokinase have been shown to be useful adjuncts to chest tube drainage in the treatment of complicated pleural effusion and empyema, there have been no reports on the use of intrapleural alteplase. This report demonstrates that intrapleural administration of alteplase is a useful adjunct to tube drainage in resolving CPE. CONCLUSIONS: This patient's CPE resolved when intrapleural alteplase was used as an adjunct to chest tube drainage and antibiotics. Controlled trials need to be conducted to investigate fully the efficacy, dosing, and safety of intrapleural alteplase in the treatment of patients with CPE and empyema. PMID- 12639167 TI - Possible captopril-induced toxic epidermal necrolysis. AB - OBJECTIVE: To report a case of toxic epidermal necrolysis (TEN) possibly induced by captopril. CASE SUMMARY: An 11-week-old boy was referred to our hospital for tetrology of fallot surgical repair, performed on admission day 2. On day 3, the patient developed third-degree heart block, necessitating pacemaker connection and oral theophylline 3 mg/kg 3 times daily. Captopril 1 mg orally twice daily, intravenous furosemide 7 mg every 12 hours, and oral aldactone 7 mg twice daily were started. On day 5, the patient developed scaling erythematous skin lesions. On day 7, his temperature spiked to 37.8 degrees C, and pus discharge from the pacing wire site was noticed. Intravenous vancomycin 80 mg 3 times daily and intravenous ceftazidime 200 mg 3 times daily were initiated. On the same day, captopril was discontinued because we suspected that it had induced the skin reaction. On day 15, the infant's skin problem progressed. The dermatologist diagnosed partial TEN. On that day, theophylline and furosemide were also discontinued. On day 16, the patient still had some blisters, but the skin started to show signs of healing, until complete healing occurred on day 22. The infant was discharged on oral medications: furosemide 7 mg twice daily, aldactone 7 mg twice daily, and enalapril 0.1 mg twice daily. Three weeks later, he was followed up. No recurrences were observed. CONCLUSIONS: This case suggests captopril induces TEN when combined with other sulfonamide medications. An objective causality assessment revealed that the adverse drug event was possible. Although it is a rare complication, healthcare providers should be familiar with its potential to occur and take appropriate treatment and prevention measures. PMID- 12639168 TI - Psychosis associated with interferon alfa therapy for chronic hepatitis B. AB - OBJECTIVE: To report a case of persistent psychosis that developed during interferon alfa (IFN-alpha) therapy for chronic hepatitis B. CASE SUMMARY: A 26 year-old man who was diagnosed with active chronic hepatitis B began treatment with IFN-alpha. Five months after initiation of therapy, he developed acute psychosis with prominent persecutory delusions and auditory hallucinations. Despite discontinuation of IFN-alpha therapy and addition of antipsychotic drug treatment, only partial recovery from psychosis was observed after 4 months of hospitalization. DISCUSSION: Unlike many previously reported cases, this patient showed only partial recovery from psychosis, despite the discontinuation of IFN therapy. Except for receiving a relatively high dose of IFN-alpha (10 million units 3 times/wk), the patient did not have any previously proposed risk factors for developing psychiatric adverse effects. The Naranjo probability scale indicates a probable relationship between the acute psychosis and IFN therapy. CONCLUSIONS: Despite its rare occurrence, psychosis can emerge during IFN-alpha therapy. This adverse effect may persist for several months, even after appropriate medical management. IFN-alpha should be used with careful monitoring of patients' psychiatric status during all stages of therapy. PMID- 12639169 TI - Combination risperidone and SSRI-induced serotonin syndrome. AB - OBJECTIVE: To report 2 cases of serotonin syndrome associated with combined therapy of risperidone and selective serotonin-reuptake inhibitors (SSRIs) in elderly patients. CASE SUMMARIES: An 86-year-old white man was admitted to the emergency department because of increased confusion and generalized weakness over the past several days. His medication history indicated paroxetine 10 mg/d and risperidone 0.25 mg/d. The patient's confusion worsened and underwent acute changes that resembled delirium. He was placed in a geri chair and he became extremely agitated. He was then treated with escalating doses of risperidone. The patient died on day 5 of admission, at which time he was being treated with risperidone 2-3 mg/d. A 78-year-old white female nursing home resident was admitted to the emergency department because of increased confusion and generalized weakness. She was being treated with paroxetine for depression and risperidone for agitation. Her risperidone dose was increased to manage agitation. The patient's agitation worsened with increasing doses of risperidone; she developed tremor, dizziness, and muscle incoordination. After psychopharmacologic consultation, the risperidone and paroxetine were discontinued and she was treated with lorazepam. The patient recovered, returned to baseline status in 2 days, and was later transferred back to the nursing home. DISCUSSION: We believe that in both cases, serotonin syndrome was precipitated by risperidone in combination with SSRI antidepressants. A literature search indicated one report of serotonin syndrome with a combination of risperidone and paroxetine. CONCLUSIONS: An objective causality assessment revealed that the adverse drug event was probable in the first patient and definite in the second patient. We caution clinicians treating elderly patients with combined risperidone and SSRIs to include serotonin syndrome in differential diagnosis if the patient is showing signs of increasing agitation with escalating doses of risperidone. PMID- 12639170 TI - Aztreonam treatment of Pasteurella multocida cellulitis and bacteremia. AB - OBJECTIVE: To report a case of Pasteurella multocida cellulitis and bacteremia treated successfully with aztreonam. CASE SUMMARY: An 81-year-old white man with multiple antibiotic allergies was admitted with severe cellulitis of the left arm and bacteremia due to P. multocida. The patient was treated for 14 days with aztreonam and had complete resolution of the infection. DISCUSSION: To our knowledge, this is the first published case describing successful treatment of P. multocida cellulitis and bacteremia with aztreonam. Antimicrobials recommended for use in P. multocida infections include penicillin, ampicillin, amoxicillin, second- and third-generation cephalosporins, tetracyclines, fluoroquinolones, and trimethoprim/sulfamethoxazole. There is very little information in the current literature regarding the activity of aztreonam toward P. multocida. CONCLUSIONS: This case demonstrates the potential use of aztreonam for P. multocida cellulitis and bacteremia in those instances where antibiotics of choice cannot be given. PMID- 12639171 TI - Recurrent cholestasis due to ampicillin. AB - OBJECTIVE: To present a single case of ampicillin-induced recurrent cholestasis and a literature review. CASE SUMMARY: A 23-year-old man was hospitalized due to recurrent and self-limited cholestatic symptoms. He had used ampicillin before each cholestatic attack. He became well clinically and biochemically each time after cessation of the drug. One year after his recovery and discontinuance of ampicillin, the patient has had no recurrence of cholestasis. An objective causality assessment revealed that the adverse drug reaction was probable. DISCUSSION: Ampicillin-related hepatotoxicity is very rare, with injury being mainly hepatocellular. To our knowledge, there is only 1 case report in the literature referring to chronic cholestatic-type hepatotoxicity related to ampicillin. CONCLUSIONS: Ampicillin, which is one of the most widely used antibiotics, may cause recurrent cholestatic hepatitis. Clinicians should be aware of this adverse effect, and it should be kept in mind during diagnostic workup of liver injury. PMID- 12639172 TI - Possible anaphylaxis after propofol in a child with food allergy. AB - OBJECTIVE: To report a case of anaphylaxis due to propofol in a child with allergies to egg and peanut oil. CASE SUMMARY: A 14-month-old boy with a history of reactive airway disease was hospitalized for treatment of respiratory symptoms. The patient had documented allergies to egg, peanut oil, and mold. Within the first few hours after admission, acute respiratory decompensation occurred, and arrangements were made to transfer the patient to our tertiary-care hospital. Prior to transfer, he was emergently intubated under sedation and paralysis with propofol and rocuronium. When emergency air transport arrived, the patient was hypotensive and tachycardic. His symptoms of anaphylaxis were managed throughout the flight and, upon arrival at our institution, the patient was admitted to the Pediatric Intensive Care Unit. He improved over a 5-day hospital course, and his caregivers were instructed to avoid propofol in the future. The patient's anaphylactic reaction following propofol was rated as a possible adverse drug reaction using the Naranjo probability scale. DISCUSSION: The use of propofol in pediatric patients for procedural sedation has gained increased favor. Since the propofol formulation contains both egg lecithin and soybean oil, its use is contraindicated in patients with hypersensitivities to these components. Several other drugs have a food component, resulting in contraindications and warnings in product labeling. CONCLUSIONS: Propofol should be avoided in patients with allergies to egg and/or soybean oil, if possible. Clinicians should consider the potential for adverse drug events in patients with select food allergies. PMID- 12639173 TI - Venous thromboembolism prevention with LMWHs in medical and orthopedic surgery patients. AB - OBJECTIVE: To discuss the role of low-molecular-weight heparins (LMWHs) in the prevention of venous thromboembolism (VTE) in medical and orthopedic surgery patients. VTE prophylaxis trials in these practice settings establishing the current use of LMWHs marketed in the US are included. An overview is also provided of VTE incidence, risk factors, and prophylaxis consensus guidelines. DATA SOURCES AND STUDY SELECTION: Clinical trials, review articles, and meta analyses for Food and Drug Administration-approved LMWHs were identified from a MEDLINE search (1980-March 2002). Search terms included dalteparin, enoxaparin, internal medicine, low-molecular-weight heparin, orthopedic surgery, risk factors, tinzaparin, and venous thromboembolism. DATA SYNTHESIS: Consensus guidelines are useful as an initial guide to appropriate VTE prophylaxis; however, a review of the primary literature is needed to identify optimal agents, regimens, or interventions. LMWHs have demonstrated sound efficacy in VTE prevention; however, the quantity and quality of literature are not always comparable for the available agents. CONCLUSIONS: Enoxaparin has demonstrated efficacy and safety in VTE prevention in medical patients, whereas information is limited or lacking for dalteparin and tinzaparin. Total hip replacement (THR) trials have been conducted with all US-marketed LMWHs and have demonstrated the efficacy and safety of each agent. Trials specifically establishing the efficacy of an LMWH in total knee replacement surgery (TKR) have been published for enoxaparin. One combination THR and TKR trial has been published for tinzaparin. These trial outcomes have positioned the LMWHs as key alternatives to adjusted dose warfarin for VTE prophylaxis in orthopedic surgery. Inherent differences between LMWHs prevent the extrapolation of clinical outcomes from 1 trial to another. PMID- 12639174 TI - Ramipril for the prevention and treatment of cardiovascular disease. AB - OBJECTIVE: To evaluate the effectiveness of ramipril in the prevention and treatment of cardiovascular disease and determine its need for inclusion on a formulary. DATA SOURCES: A MEDLINE and PubMed database search was conducted (1987 May 2002). Only journals written in the English language were selected for review. DATA EXTRACTION AND STUDY SELECTION: Articles reporting the use of ramipril in humans were evaluated. Emphasis was placed on randomized, controlled trials assessing efficacy. DATA SYNTHESIS: Ramipril is an angiotensin-converting enzyme (ACE) inhibitor that exerts its effects through inhibition of the renin angiotensin-aldosterone system. It exhibits a safety profile that is similar to that of other ACE inhibitors and is comparable in cost to the majority of the available agents. Clinical trials have proven the effectiveness of ACE inhibitors in the treatment of hypertension, heart failure, and nephropathy. Ramipril, however, is the only ACE inhibitor currently approved for the prevention of cardiovascular events in high-risk patients without evidence of left-ventricular dysfunction or heart failure, based on the results of the HOPE (Heart Outcomes Prevention Evaluation) trial. Whether this effect is specific to ramipril has yet to be proven. This article emphasizes the major trials involving ramipril including the AIRE (Acute Infarction Ramipril Efficacy), REIN (Ramipril Efficacy in Nephropathy), and HOPE trials. CONCLUSIONS: Although similar to other ACE inhibitors in many aspects, it cannot be assumed that the benefits shown with ramipril in the HOPE trial are a class effect. Ongoing trials should help to clarify this matter. Until this time, current evidence justifies the inclusion of ramipril on a formulary. PMID- 12639175 TI - Voriconazole: a new triazole antifungal agent. AB - OBJECTIVE: To review the pharmacology, in vitro susceptibility, pharmacokinetics, clinical efficacy, and adverse effects of voriconazole, a triazole antifungal agent. DATA SOURCES: A MEDLINE search, restricted to English language, was conducted from 1990 to June 2002. Supplementary sources included program abstracts from the Interscience Conference on Antimicrobial Agents and Chemotherapy and the Infectious Diseases Society of America from 1996 to 2001 and manufacturer information available through the Food and Drug Administration's Web site. DATA EXTRACTION: All published and unpublished trials and abstracts citing voriconazole were selected. DATA SYNTHESIS: Voriconazole has shown in vitro activity against many yeasts and a variety of mold and dermatophyte isolates. Voriconazole can be administered either orally or parenterally. It exhibits good oral bioavailability, wide tissue distribution including distribution into the central nervous system, and hepatic metabolism. Drug interactions occur through inhibition of the CYP2C9, CYP2C19, and CYP3A4 isoenzymes, resulting in alterations in kinetic parameters of either voriconazole or the interacting agent. Efficacy has been illustrated in open, noncomparative studies of aspergillosis in immunocompromised patients. Human case reports describe successful treatment of rare fungal pathogens. The most commonly reported adverse events include visual disturbances and elevations in liver function tests. CONCLUSIONS: Voriconazole is at least as effective as amphotericin B in the treatment of acute invasive aspergillosis in immunocompromised patients. It has similar efficacy as fluconazole in treatment of esophageal candidiasis. Voriconazole did not achieve statistical non-inferiority to liposomal amphotericin B for empirical therapy in patients with neutropenia and persistent fever, diminishing enthusiasm for use in this indication until additional trials are completed. Based on case reports and in vitro efficacy, voriconazole may prove to be a clinically useful agent in the treatment of other fungal disease. PMID- 12639176 TI - Intravenous proton-pump inhibitors versus H2-antagonists for treatment of GI bleeding. AB - OBJECTIVE: To evaluate the evidence supporting the use of intravenous proton-pump inhibitors in the treatment of gastrointestinal (GI) hemorrhage in comparison with histamine(2) (H(2))-receptor antagonists. DATA SOURCES: Clinical literature was accessed through a MEDLINE search (1966-October 2002). Data from abstracts and fully published articles were retrieved for analysis. Key search terms included pantoprazole, omeprazole, proton-pump inhibitors, gastrointestinal hemorrhage, histamine(2)-receptor antagonists, ranitidine, and cimetidine. DATA SYNTHESIS: There are limited published clinical outcome data evaluating the use of intravenous pantoprazole in patients with upper GI hemorrhage. However, there are several gastric pH studies suggesting that intravenous pantoprazole is effective in quickly obtaining and maintaining a pH >6. When considering the results from studies of high-dose intravenous omeprazole, in addition to the pantoprazole data, the relative efficacy of intravenous proton-pump inhibitors appears to be superior to that of intravenous H(2)-receptor antagonists in providing a more predictable and sustained pH control. CONCLUSIONS: Intravenous proton-pump inhibitors are suitable, possibly superior, alternatives to intravenous H(2)-receptor antagonists in treatment of upper GI bleeding. PMID- 12639177 TI - Safety of selegiline with cold medications. AB - OBJECTIVE: To evaluate the safety of the coadministration of selegiline with cold medications. DATA SOURCES: Clinical literature accessed through MEDLINE (1965 September 2002), IPA database, and Drug-Reax System. The following search terms were used: selegiline, pseudoephedrine, dextromethorphan, MAOI, and drug interactions. Somerset Pharmaceuticals, the marketers of Eldepryl (selegiline HCl), were also contacted. DATA SYNTHESIS: Despite a warning against its concomitant use with pseudoephedrine and dextromethorphan, interactions with selegiline have not been reported. However, there have been reports of patients experiencing adverse events with related agents. CONCLUSIONS: Patients taking selegiline should try to avoid pseudoephedrine and dextromethorphan or use drugs without interaction potential. If selegiline is used with these medications, watch for adverse events or replace selegiline with another drug. PMID- 12639178 TI - Effect of cyclooxygenase-2 inhibitors on blood pressure. AB - OBJECTIVE: To evaluate the effect of cyclooxygenase-2 selective inhibitors (CSIs) on blood pressure. DATA SOURCES: Clinical literature accessed through MEDLINE (1966-May 2002). Key search terms included COX-2 selective inhibitors; anti inflammatory agents, nonsteroidal; celecoxib; rofecoxib; and hypertension. DATA SYNTHESIS: Data from prospective studies on the effects of CSIs on blood pressure are conflicting. Several studies have reported increased blood pressure as an adverse effect of CSIs. CONCLUSIONS: Additional studies are needed to evaluate the effects of CSIs on blood pressure. CSIs should be used with caution in hypertensive patients and blood pressure monitored closely if a CSI is indicated. PMID- 12639179 TI - Pharmacoeconomic implications of large multicenter clinical trials. PMID- 12639180 TI - Atypical tuberculosis skin test reaction. PMID- 12639181 TI - Mirtazapine in suicidal Huntington's disease. PMID- 12639182 TI - Moxifloxacin-induced Clostridium difficile diarrhea. PMID- 12639183 TI - Comment: delayed wound healing with sirolimus after liver transplant. PMID- 12639185 TI - Comment: patient-specific factors predictive of warfarin dosage requirements. PMID- 12639191 TI - Smallpox vaccination and adverse reactions: guidance for clinicians. PMID- 12639193 TI - No surprises in blood pressure awareness study findings: we can do a better job. PMID- 12639194 TI - Safety and statin therapy: reconsidering the risks and benefits. PMID- 12639195 TI - Should routine screening for prostate-specific antigen be recommended? PMID- 12639196 TI - An argument against routine prostate cancer screening. PMID- 12639199 TI - Differential effects of lipid-lowering therapies on stroke prevention: a meta analysis of randomized trials. AB - BACKGROUND: Previous overviews suggested that hydroxymethyl glutaryl coenzyme A reductase inhibitors (statins), but not other lipid-lowering therapy (LLT), may reduce stroke incidence in coronary patients. OBJECTIVE: To investigate the amplitude and sources of heterogeneity of LLT effects on stroke prevention. METHODS: We searched the literature from 1966 to 2001 and then conducted a meta analysis including randomized trials of primary and secondary coronary heart disease prevention, testing statins, nonstatin drugs, diet, or other interventions and providing data on stroke incidence. RESULTS: The meta-analysis (38 trials, 83 161 patients, mean follow-up of 4.7 years) showed a significant relative risk reduction (RRR) of strokes by LLT of 17% (P<.001), without significant heterogeneity between trials and between subgroups according to either the type of prevention (primary or secondary) or the type of LLT. The most substantial effects were obtained, however, with statins (RRR, 26%). Effect model analysis showed that treatment benefit appeared constant whatever the risk of stroke, suggesting that LLT may be effective in a population with a higher risk of stroke. Weighted regression showed a significant correlation between RRR of stroke and total cholesterol levels (baseline, final, and change). Only final cholesterol allowed clear separation between benefit (RRR>0) and no effect (RRR<0) of LLT on stroke incidence, with a cutoff for benefit of 232 mg/dL (6.0 mmol/L). CONCLUSION: Lipid-lowering therapy reduces stroke incidence in coronary patients, especially when total cholesterol level is lowered to less than 232 mg/dL (6.0 mmol/L), which explains the best results being obtained with statins. PMID- 12639200 TI - Awareness, knowledge, and attitudes of older americans about high blood pressure: implications for health care policy, education, and research. AB - BACKGROUND: The prevalence of high blood pressure (HBP), particularly isolated systolic hypertension, rises with age, whereas control rates decline. METHODS: Since awareness, knowledge, and attitudes about HBP can influence control, information on these factors was obtained by telephone interview of a nationally representative sample of 1503 adults 50 years or older. RESULTS: Among Americans 50 years or older, 94% had at least one blood pressure (BP) measurement during the past year, yet 46% did not know their BP. Only 27% acknowledged current HBP, although 37% reported taking antihypertensive medications. Systolic hypertension was probably underrecognized, since 30% who reported a value of 140 mm Hg or higher indicated they did not have HBP. Among those acknowledging current HBP, 80% reported taking medications "precisely as prescribed." Of the approximately 20% of hypertensive patients no longer taking medications or taking fewer medications than prescribed, cost was a major factor in approximately 1 in 5 or roughly 4% of the total. Sixty percent of patients receiving treatment indicated that medications alone do not control HBP. Most survey respondents (>or=90%) concurred that several lifestyle changes lower BP; 75% reported a lifestyle change; and 61% indicated it lowered their BP. When asked what HBP information was most important, 34% reported alternative therapies and 28% reported prevention strategies. CONCLUSIONS: Limited awareness of systolic hypertension emerges as a greater barrier to BP control than cost of medications in Americans 50 years or older. Many older Americans prefer to integrate traditional, complementary, and alternative strategies. Education addressing limited awareness of systolic hypertension, policies facilitating a more holistic management approach, and research identifying the most effective innovations may improve outcomes. PMID- 12639201 TI - Screening for statin-related toxicity: the yield of transaminase and creatine kinase measurements in a primary care setting. AB - BACKGROUND: Recommendations for monitoring levels of transaminases (alanine aminotransferase and aspartate aminotransferase) and of creatine kinase (CK) in patients taking 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) vary and are not based on data from clinical practice. We performed a study to determine the yield of routine screening of aminotransferase and CK levels among patients taking statins. METHODS: We performed a retrospective review of a primary care practice's computerized medical record. A computerized search identified all patients with a statin on their medication list and gave their alanine aminotransferase, aspartate aminotransferase, and CK values for 1998. We reviewed the records of all patients for whom these values were significantly or moderately abnormal to determine the values' relationship to statin therapy and outcomes. RESULTS: During the year of the study, 1014 (85%) of the 1194 patients who had a statin on their medication list had at least 1 monitoring test performed. Of these 1014 patients, 10 (1.0%) had a significant elevation and 5 (0.5%) a moderate elevation of transaminase levels, but none of these abnormalities appeared to be related to statin use. Moreover, 6 (0.9%) patients had at least 1 significantly abnormal CK value but it did not appear to be attributable to a statin; and of the 14 (2.1%) patients who had a moderate CK elevation, it was potentially due to a statin in only 2. There were no documented adverse sequelae associated with these abnormal results. CONCLUSIONS: In this study of statin use in a primary care practice, routine monitoring revealed no cases of significantly or moderately abnormal transaminase values attributable to statins. No significantly abnormal and only 2 moderately abnormal CK values were potentially attributable to statin use. This study questions the usefulness of routine measurement of transaminase and CK levels in all patients taking statins. PMID- 12639202 TI - Anticoagulation of pregnant women with mechanical heart valves. AB - The management of pregnant women with mechanical heart valves is challenging. Recently, based on small numbers of patients and poor-quality data, correspondence from Aventis Pharmaceuticals Inc has described treatment "failures" and concerns about teratogenicity with the use of the low-molecular weight heparin (LMWH) enoxaparin. The company issued a "Warning" that enoxaparin should not be used in patients with prosthetic heart valves and a "Precaution" about potential teratogenicity. This has created a huge problem for physicians managing pregnant women with prosthetic heart valves because the alternatives, unfractionated heparin and warfarin, are problematic. There have been case reports of failures (including death from thrombosed valves) with unfractionated heparin, whereas the package insert for warfarin states that the drug is contraindicated during pregnancy because of potential teratogenicity. Initially, LMWHs appeared suitable for pregnant patients with prosthetic heart valves. Unfortunately, the company correspondence, presumably supported by the Food and Drug Administration (FDA), raises medicolegal concerns with use of any LMWH. We believe that pharmaceutical companies and the FDA should not endorse scientifically unsupported claims that eliminate acceptable therapeutic options. This correspondence has created considerable confusion among patients and treating physicians and is likely to lead to frivolous lawsuits and preclude the performance of properly designed trials in pregnant women. We believe a consensus conference among experts in the field to identify key unresolved issues and a commitment by the FDA and industry to perform appropriate studies are now critical. PMID- 12639203 TI - Echinacea and truth in labeling. AB - BACKGROUND: Echinacea sales represent 10% of the dietary supplement market in the United States, but there is no guarantee as to the content, quality, variability, or contamination in Echinacea preparations. OBJECTIVE: To qualitatively and quantitatively assess the contents of Echinacea-only preparations available in a retail setting. METHODS: One of each single-herb Echinacea preparations that were available in August 2000 was obtained from several stores in the Denver, Colo, area. Thin-layer chromatography (TLC) was used to determine species and measure quantity. From this information, accuracy of species labeling and comparison of constituent to labeled content were assessed. The samples were stratified by whether they were labeled as standardized, and the standardized and nonstandardized samples were compared by ratio of constituent to labeled content. RESULTS: Of the samples, 6 (10%) of 59 preparations contained no measurable Echinacea. The assayed species content was consistent with labeled content in 31 (52%) of the samples. Of the 21 standardized preparations, 9 (43%) met the quality standard described on the label. Labeled milligrams were weakly associated with measured constituent (r = 0.35; P =.02). CONCLUSIONS: Echinacea from retail stores often does not contain the labeled species. A claim of "standardization" does not mean the preparation is accurately labeled, nor does it indicate less variability in concentration of constituents of the herb. PMID- 12639204 TI - Mortality and cancer incidence among individuals with Down syndrome. AB - BACKGROUND: Individuals with Down syndrome (DS) have a predisposition to leukemia and possibly other cancers and excess mortality from other conditions, but information on the magnitude of risk associated with specific cancers or causes of death is sparse. METHODS: Mortality experience and cancer incidence were evaluated in a combined cohort of 4872 individuals with a hospital discharge diagnosis of DS in Sweden (1965-1993) or Denmark (1977-1989) by linkage to national cancer and vital statistics registries. Standardized incidence ratios (SIRs) and standardized mortality ratios (SMRs) were estimated by comparison with age, sex, and calendar-year expected values. RESULTS: Individuals with DS had an increased risk of incident acute lymphocytic (SIR, 24.2; 95% confidence interval [CI], 15.2-36.6; n = 22) and acute nonlymphocytic (SIR, 28.2; 95% CI, 15.7-48.3; n = 14) leukemias. Risks of testicular cancer (SIR, 3.7; 95% CI, 1.0-9.4; n = 4) and liver cancer (SIR, 6.0; 95% CI, 1.2-17.5; n = 3) were also elevated. Individuals with DS also experienced elevated mortality attributed to stomach cancer (SMR, 6.4; 95% CI, 1.7-16.4; n = 4), dementia and Alzheimer disease (SMR, 54.1; 95% CI, 27.9-94.4), epilepsy (SMR, 30.4; 95% CI, 13.9-57.7), ischemic heart disease (SMR, 3.9; 95% CI, 2.7-5.4), other heart disease (SMR, 16.5; 95% CI, 11.0 23.7), cerebrovascular disease (SMR, 6.0; 95% CI, 3.5-9.6), infectious diseases (SMR, 12.0; 95% 6.0-21.4), and congenital anomalies (SMR, 25.8; 95% CI, 21.0 31.4). CONCLUSIONS: Individuals with DS have a substantially increased risk of mortality due to specific causes and may have an elevated risk of other incident cancers in addition to leukemia. These results provide clues regarding chromosome 21 gene involvement in diseases that complicate DS and are important for disease detection and care of affected individuals. PMID- 12639205 TI - Prevalence of current DSM-IV alcohol use disorders in short-stay, general hospital admissions, United States, 1994. AB - BACKGROUND: This study provides, to our knowledge, the first national prevalence estimates of Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV), alcohol use disorders based on a structured, diagnostic instrument for inpatient admissions to US general hospitals. Existing prevalence estimates for inpatient admissions came from studies conducted in 1 or 2 hospitals and therefore do not support national inference. METHODS: A multistage probability sample was designed to represent acute care admissions to nonfederal, short-stay, general hospitals in the contiguous United States; 2040 admissions (1613 males and 427 females) in 90 hospitals participated. RESULTS: An estimated 1.8 million (95% confidence interval, 1.3-2.2 million) annual hospital admissions met the criteria for a current (ie, in the past 12 months) DSM-IV alcohol use disorder. Overall prevalence was estimated to be 7.4% (95% confidence interval, 5.6%-9.1%). Among current-drinking admissions, estimated prevalence was 24.0% (95% confidence interval, 18.7%-29.4%), and males and females had similar rates. Pairwise comparisons showed significant elevations in the prevalence of alcohol use disorders in current-drinking admissions who were younger, black, unmarried, of a lower socioeconomic status, on Medicaid or without health insurance, smokers, or drug users. Prevalence of alcohol use disorders was also significantly higher in current-drinking admissions in hospitals that were government owned, had medical school affiliations, or had a high number of emergency department visits per day. CONCLUSIONS: The prevalence of alcohol abuse or dependence in current-drinking admissions was substantial, suggesting that hospitalization offers a unique opportunity to identify alcohol use disorders. Further research is needed to determine factors that may be associated with significant pairwise results, especially for race or ethnicity. We recommend alcohol screening of all hospitalized drinkers, followed, as appropriate, by diagnostic evaluation and referral or intervention. PMID- 12639206 TI - Effect of 1-year oral administration of dehydroepiandrosterone to 60- to 80-year old individuals on muscle function and cross-sectional area: a double-blind placebo-controlled trial. AB - BACKGROUND: The age-related decline of dehydroepiandrosterone and its sulfate ester levels is thought to be related to the development of age-associated usual modifications, such as neuromuscular function impairments. It is often claimed that individuals can enhance their muscular capacity by boosting dehydroepiandrosterone levels through oral supplementation. However, to our knowledge, there have been no controlled studies on a significant number of individuals demonstrating positive effects on the neuromuscular system. This study determines if 1-year administration of a replacement dose of dehydroepiandrosterone, 50 mg/d, orally administered, could have a beneficial influence on several determinants of the muscular function altered during aging. METHODS: This work was completed within the frame of the DHEAge Study, which was conducted in France from March 1, 1998, to October 31, 1999. It was performed on 280 healthy ambulatory and independent men and women aged 60 to 80 years. The study design was a double-blind placebo-controlled trial. Dehydroepiandrosterone sulfate serum concentration, handgrip strength, isometric and isokinetic knee muscle strength, and thigh (fat and muscle) cross-sectional area were analyzed before and just after 12 months of placebo or dehydroepiandrosterone treatment. RESULTS: The results give evidence that dehydroepiandrosterone administration restores dehydroepiandrosterone sulfate serum concentrations to the normal range for young adults (aged 20-50 years). However, no positive effect inherent to dehydroepiandrosterone treatment was observed either on muscle strength or in muscle and fat cross-sectional areas. CONCLUSIONS: The compensation of the deficit of dehydroepiandrosterone during aging using a 50-mg/d dose does not induce beneficial effects on muscle state in healthy subjects. The conditions in which dehydroepiandrosterone could contribute to preserve or improve muscle strength and morphological features still need to be determined. PMID- 12639207 TI - A cohort study of the incidence of serious acute liver injury in diabetic patients treated with hypoglycemic agents. AB - BACKGROUND: The incidence of acute liver failure or serious liver injury in diabetic patients is needed to evaluate the safety of hypoglycemic drug therapy. METHODS: We conducted a retrospective cohort study of 5 health maintenance organizations. Study patients were 171,264 health plan members 19 years or older when they received oral hypoglycemic drugs or insulin between April 1, 1997, and June 30, 1999. We searched for hospital discharge diagnoses and procedures potentially indicative of acute liver injury and reviewed the full-text medical records. Acute liver failure was defined as acute liver disease and (1) hepatic encephalopathy, (2) prothrombin time prolongation greater than 3 seconds or international normalized ratio greater than 1.5, and (3) a total bilirubin level greater than 3.0 mg/dL (>51 micro mol/L). Acute liver injury was diagnosed in individuals who did not meet 1 or more of the criteria for acute liver failure but had alanine transaminase or aspartate transaminase levels greater than 500 U/L. RESULTS: We identified 35 cases of acute liver failure or injury not clearly attributable to a known cause other than use of hypoglycemic agents. The age- and sex-standardized incidence per 1000 person-years was 0.15 for insulin users, 0.08 for sulfonylurea users, 0.12 for metformin users, and 0.10 for troglitazone users. The incidence was higher (on the order of 0.3 per 1000) during the first 6 months of exposure to all hypoglycemic agents. CONCLUSIONS: Acute liver failure or injury not clearly attributable to other known causes occurred on the order of 1 per 10,000 person-years among diabetic patients treated with oral hypoglycemic drugs or insulin. PMID- 12639208 TI - Thrombocytopenia associated with levodopa treatment. PMID- 12639209 TI - End-of-life care and congestive heart failure. PMID- 12639210 TI - Frailty is a strong modulator of heart failure-associated mortality. PMID- 12639212 TI - HIV sexual risk reduction intervention with heterosexual men. PMID- 12639213 TI - Is MRSA more pathogenic in critically ill patients? PMID- 12639215 TI - Therapeutic role of vasopressin receptor antagonism in patients with liver cirrhosis. AB - Vasopressin, or antidiuretic hormone, is a peptide hormone that is released from the posterior pituitary gland in response to changes in blood pressure and plasma osmolality. The main pathophysiological states associated with high plasma vasopressin concentrations are cirrhosis, cardiac failure and syndrome of inappropriate antidiuretic hormone (SIADH) secretion. Pharmacological treatments for disorders of excess vasopressin secretion have been limited. However, oral bio-available selective and non-selective V(1) and V(2) receptor antagonists have recently become available for clinical use. Water retention in cirrhosis is a common problem, leading to ascites, peripheral oedema and hyponatraemia. Raised plasma vasopressin concentrations and decreased delivery of glomerular filtrate are believed to be the most important factors in the development of water retention. V(2) receptor antagonists are aquaretic agents that promote water excretion and improve hyponatraemia. Their potential role in cirrhosis has been examined in a number of recent studies that have shown increased free water clearance and serum sodium concentrations with few adverse effects. V(2) receptor antagonists represent a novel and promising new class of agent that may have major clinical utility in the treatment of patients with liver cirrhosis. PMID- 12639216 TI - The C242T p22phox polymorphism and endothelium-dependent vasodilation in subjects with hypercholesterolaemia. AB - Oxidative stress plays a major pathogenetic role in cardiovascular disease. The C242T variant of the CYBA gene encoding the p22phox subunit of the NAD(P)H oxidase, a major source of superoxide production, has been shown to be associated with coronary artery disease and with vascular superoxide production in human veins ex vivo. Since superoxide degrades nitric oxide (NO), we hypothesized that the C242T variant influences endothelium-dependent vasodilation of the human forearm vasculature in vivo. In the present study, 90 subjects with elevated cholesterol levels were stratified for the C242T polymorphism of the CYBA p22phox gene. Endothelium-dependent and -independent vasodilation were assessed by plethysmographic monitoring of forearm blood flow responses to intra-arterial infusion of acetylcholine and sodium nitroprusside respectively. N(G)-Monomethyl L-arginine (L-NMMA) was infused to analyse NO-mediated basal vascular tone. Baseline parameters (age, gender, blood pressure, body mass index, cholesterol level) were similar across the genotypes. No differences in forearm blood flow responses to the intra-arterial infusion of acetylcholine, sodium nitroprusside or L-NMMA were found across the CYBA p22phox genotypes. Our sample size of n =90 had a power of >80% (beta=0.20) with a P value of <0.05 (alpha=0.05) to detect a difference greater than 156% in the forearm blood flow response to acetylcholine across genotypes (S.D. 336%; average increase in forearm blood flow=514%). In conclusion, at a power of 80%, our study excludes a major effect of the C242T CYBA p22phox polymorphism on acetylcholine-mediated endothelium-dependent vasodilation and basal NO-mediated vascular tone of the human forearm circulation in subjects with hypercholesterolaemia. PMID- 12639217 TI - Decrease of serum levels of the anti-inflammatory cytokine interleukin-10 in patients with advanced chronic heart failure. AB - Inflammation plays a significant contributory role in the pathogenesis of chronic heart failure (CHF). Many studies have shown enhanced plasma levels of proinflammatory cytokines [i.e. tumour necrosis factor-alpha (TNF-alpha) and interleukin (IL)-6] in patients with CHF. However, there are only few reports on the regulation of anti-inflammatory cytokines such as IL-10. IL-10 has potent deactivating properties in macrophages and T-cells and thus acts as a down regulator of cell-mediated immune responses. The aim of the present study was to assess whether serum concentrations of IL-10 significantly differ between patients with CHF and healthy control subjects. Patients with CHF [ n =50; 66.9+/ 12.6 years; mean ejection fraction, 22.1+/-9.2%; New York Heart Association (NYHA) class II-IV] and 25 healthy controls (63.6+/-10.2 years) were examined. Of the 50 patients with CHF, 32 patients were taking aspirin (100 mg/day) and 33 patients had lipid-lowering therapy with a statin. Serum IL-10 as well as TNF alpha concentrations were measured using commercially available immunoassays. Patients with CHF showed significantly lower IL-10 concentrations (2.3+/-1.9 compared with 5.2+/-2.3 pg/ml; P <0.001). Patients with advanced CHF (NYHA class III and IV) had the lowest IL-10 plasma levels. Aspirin and statin therapy did not significantly influence serum levels of IL-10. The ratio of TNF-alpha to IL 10 was significantly higher in patients with advanced CHF (NYHA class III and IV, ratio 3.2+/-1.2 and 3.1+/-1.1 respectively, compared with control 0.4+/-0.2; P <0.01). Our present study demonstrates significantly decreased serum levels of IL 10 in patients with advanced CHF. Since IL-10 is known as a potent anti inflammatory cytokine, its decrease in advanced CHF may favour the inflammatory milieu in CHF. PMID- 12639218 TI - Decreased left ventricular longitudinal contraction in normotensive and normoalbuminuric patients with Type II diabetes mellitus: a Doppler tissue tracking and strain rate echocardiography study. AB - Type II diabetes mellitus is associated with congestive heart failure with preserved ejection fraction. This group of patients has been assumed to have isolated diastolic dysfunction; however, the longitudinal systolic contraction of the left ventricle has not been studied previously. The objective of the present study was to investigate the longitudinal contraction of the left ventricle in normotensive Type II diabetes mellitus patients with normal ejection fraction. We examined 32 normotensive patients with Type II diabetes mellitus with ejection fraction >0.55 and fractional shortening >0.25. Exclusion criteria were angina pectoris, cardiac valve disease, albuminuria, retinopathy or neuropathy. Normal subjects (n =32) served as controls. A 16 segment model of motion amplitude assessed left ventricular longitudinal contraction and the average of the segments was calculated as the tissue tracking score index. Peak systolic velocity and strain rate was also obtained in each segment. Patients with Type II diabetes mellitus had a significantly lower tissue tracking score index compared with normal subjects (5.8+/-1.6 mm compared with 7.7+/-1.1 mm; P <0.001). Mean peak systolic velocity was also significantly lower (4.3+/-1.5 cm/s compared with 5.4+/-1.0 cm/s; P <0.001), as well as peak systolic strain rate (-1.2+/-0.3 s(-1) compared with -1.6+/-0.4 s(-1); P <0.001). Patients with Type II diabetes mellitus and preserved diastolic function had a significantly lower tissue tracking score index compared with normal subjects (6.6+/-1.5 mm; P <0.001), but patients with diastolic dysfunction had an even more profound decrease in tissue tracking score index compared with patients without diastolic dysfunction (4.9+/ 0.9 mm; P <0.01). In conclusion, the longitudinal systolic contraction was significantly decreased in normotensive patients with Type II diabetes mellitus with normal ejection fraction, which was most profound in patients with concomitant diastolic dysfunction. PMID- 12639219 TI - Tissue inhibitor of metalloproteinases-1 signalling pathway leading to erythroid cell survival. AB - Tissue inhibitors of metalloproteinases (TIMP) are specific inhibitors of matrix metalloproteinases (MMPs) and thus participate in maintaining the balance between extracellular matrix deposition and degradation in several physio-pathological processes. Nevertheless, TIMP must be regarded as multifunctional proteins involved in cell growth, angiogenesis and apoptosis. The molecular mechanisms induced by TIMP remain largely unknown. In the present study, we provide evidence that TIMP-1 induces a significant anti-apoptotic effect in the human erythroleukaemic cell line UT-7 and in the murine myeloid cell line 32D. Using specific kinases inhibitors, we show that TIMP-1-mediated cell survival is dependent upon Janus kinase (JAK) 2 and phosphoinositide 3-kinase (PI 3-kinase) activities. By transient transfection of dominant-negative Akt in UT-7 cells, we demonstrate that this kinase is crucial for the TIMP-1 anti-apoptotic effect. Moreover, TIMP-1 enhances specific phosphorylation of both Akt and Bad (Bcl-2/Bcl X(L)-antagonist, causing cell death) in a PI 3-kinase-dependent manner and, besides, controls the level of the anti-apoptotic protein Bcl-X(L). We conclude that TIMP-1 induces haematopoietic cell survival via the JAK2/PI 3-kinase/Akt/Bad pathway. PMID- 12639220 TI - Paraoxonase-1 promoter haplotypes and serum paraoxonase: a predominant role for polymorphic position - 107, implicating the Sp1 transcription factor. AB - Accumulating data suggest that paraoxonase-1 (PON1) is a primary determinant of the antioxidant and anti-inflammatory capacities of high-density lipoproteins (HDLs). Variations in HDLs and PON1 have been shown to influence the functions of both. There is a wide spectrum of serum PON1 mass in humans, to which promoter polymorphisms make an important contribution. The present studies attempted to define: (i) the relevance in vivo of promoter polymorphisms by analysing haplotype structure; and (ii) molecular mechanisms implicated in promoter activity. Highly significant differences (P <0.0001) in serum mass and activity were observed as a function of haplotype sequence. Of three promoter polymorphisms (-107, -824 and -907), the -107 site was shown to be of predominant importance to serum PON1. Significant increases in serum PON1 mass and activities between haplotype subgroups could be explained by unit increases in the number of high-expresser variants of the -107 site (-107C) alone. No significant contribution was observed for the -824 and -907 sites. The coding-region Leu(55)- >Met (L55M) polymorphism made an independent contribution to serum PON1 mass, which may account for variations in serum PON1 mass and activity within haplotype subgroups defined by the -107 site. A molecular basis for the effect of the -107 polymorphism on serum PON1 was indicated by the greater affinity of the high expresser variant (-107C) for hepatocyte nuclear extracts, indicating higher affinity for transcription factors. Competition studies with oligonucleotides representing the consensus (and mutated) sequence for Sp1, and the use of Sp1 antibodies, confirmed formation of complexes between the transcription factor and the PON1 promoter during incubation with nuclear extracts. The data underline the importance of the region containing the C(-107)T polymorphism for gene expression in vivo. Differences in the affinity of the -107C and -107T polymorphic fragments for nuclear extracts have been demonstrated, and coincide with their impact on gene expression. A potential role for the transcription factor Sp1 has been demonstrated, which is consistent with the disruption of an Sp1 recognition sequence by the -107 polymorphism. PMID- 12639222 TI - European Prospective Investigation into Cancer and Nutrition (EPIC): study populations and data collection. AB - The European Prospective Investigation into Cancer and Nutrition (EPIC) is an ongoing multi-centre prospective cohort study designed to investigate the relationship between nutrition and cancer, with the potential for studying other diseases as well. The study currently includes 519 978 participants (366 521 women and 153 457 men, mostly aged 35-70 years) in 23 centres located in 10 European countries, to be followed for cancer incidence and cause-specific mortality for several decades. At enrollment, which took place between 1992 and 2000 at each of the different centres, information was collected through a non dietary questionnaire on lifestyle variables and through a dietary questionnaire addressing usual diet. Anthropometric measurements were performed and blood samples taken, from which plasma, serum, red cells and buffy coat fractions were separated and aliquoted for long-term storage, mostly in liquid nitrogen. To calibrate dietary measurements, a standardised, computer-assisted 24-hour dietary recall was implemented at each centre on stratified random samples of the participants, for a total of 36 900 subjects. EPIC represents the largest single resource available today world-wide for prospective investigations on the aetiology of cancers (and other diseases) that can integrate questionnaire data on lifestyle and diet, biomarkers of diet and of endogenous metabolism (e.g. hormones and growth factors) and genetic polymorphisms. First results of case control studies nested within the cohort are expected early in 2003. The present paper provides a description of the EPIC study, with the aim of simplifying reference to it in future papers reporting substantive or methodological studies carried out in the EPIC cohort. PMID- 12639223 TI - European Prospective Investigation into Cancer and Nutrition (EPIC) calibration study: rationale, design and population characteristics. AB - The European Prospective Investigation into Cancer and Nutrition (EPIC), which covers a large cohort of half a million men and women from 23 European centres in 10 Western European countries, was designed to study the relationship between diet and the risk of chronic diseases, particularly cancer. Information on usual individual dietary intake was assessed using different validated dietary assessment methods across participating countries. In order to adjust for possible systematic over- or underestimation in dietary intake measurements and correct for attenuation bias in relative risk estimates, a calibration approach was developed. This approach involved an additional dietary assessment common across study populations to re-express individual dietary intakes according to the same reference scale. A single 24-hour diet recall was therefore collected, as the EPIC reference calibration method, from a stratified random sample of 36 900 subjects from the entire EPIC cohort, using a software program (EPIC-SOFT) specifically designed to standardise the dietary measurements across study populations. This paper describes the design and populations of the calibration sub-studies set up in the EPIC centres. In addition, to assess whether the calibration sub-samples were representative of the entire group of EPIC cohorts, a series of subjects' characteristics known possibly to influence dietary intakes was compared in both population groups. This was the first time that calibration sub-studies had been set up in a large multi-centre European study. These studies showed that, despite certain inherent methodological and logistic constraints, a study design such as this one works relatively well in practice. The average response in the calibration study was 78.3% and ranged from 46.5% to 92.5%. The calibration population differed slightly from the overall cohort but the differences were small for most characteristics and centres. The overall results suggest that, after adjustment for age, dietary intakes estimated from calibration samples can reasonably be interpreted as representative of the main cohorts in most of the EPIC centres. PMID- 12639224 TI - Overweight, obesity and fat distribution in 50- to 64-year-old participants in the European Prospective Investigation into Cancer and Nutrition (EPIC). AB - OBJECTIVE: To describe anthropometric characteristics of participants of the European Prospective Investigation into Cancer and Nutrition (EPIC). DESIGN: A cross-sectional analysis of baseline data of a European prospective cohort study. SUBJECTS: This analysis includes study populations from 25 centres in nine European countries. The British populations comprised both a population-based and a 'health-conscious' group. The analysis was restricted to 83 178 men and 163 851 women aged 50-64 years, this group being represented in all centres. METHODS: Anthropometric examinations were undertaken by trained observers using standardised methods and included measurements of weight, height, and waist and hip circumferences. In the 'health-conscious' group (UK), anthropometric measures were predicted from self-reports. RESULTS: Except in the 'health-conscious' group (UK) and in the French centres, mean body mass index (BMI) exceeded 25.0 kg m-2. The prevalence of obesity (BMI> or =30 kg m(-2)) varied from 8% to 40% in men, and from 5% to 53% in women, with high prevalences (>25%) in the centres from Spain, Greece, Ragusa and Naples (Italy) and the lowest prevalences (<10%) in the French centres and the 'health-conscious' group (UK). The prevalence of a large waist circumference or a high waist-to-hip ratio was high in centres from Spain, Greece, Ragusa and Naples (Italy) and among women from centres in Germany and Bilthoven (The Netherlands). CONCLUSIONS: Anthropometric measures varied considerably within the EPIC population. These data provide a strong base for further investigation of anthropometric measures in relation to the risk of chronic diseases, especially cancer. PMID- 12639225 TI - Physical activity of subjects aged 50-64 years involved in the European Prospective Investigation into Cancer and Nutrition (EPIC). AB - OBJECTIVE: To describe physical activity of participants in the European Prospective Investigation into Cancer and Nutrition (EPIC). DESIGN: A cross sectional analysis of baseline data of a European prospective cohort study. SUBJECTS: This analysis was restricted to participants in the age group 50-64 years, which was represented in all EPIC centres. It involved 236 386 participants from 25 centres in nine countries. In each EPIC centre, physical activity was assessed by standardised and validated questions. Frequency distribution of type of professional activity and participation in non professional activities, and age-adjusted means, medians and percentiles of time dedicated to non-professional activities are presented for men and women from each centre. RESULTS: Professional activity was most frequently classified as sedentary or standing in all centres. There was a wide variation regarding participation in different types of non-professional activities and time dedicated to these activities across EPIC centres. Over 80% of all EPIC participants engaged in walking, while less than 50% of the subjects participated in sport. Total time dedicated to recreational activities was highest among the Dutch participants and lowest among men from Malmo (Sweden) and women from Naples (Italy). In all centres, total time dedicated to recreational activity in the summer was higher than in the winter. Women from southern Europe spent the most time on housekeeping. CONCLUSIONS: There is a considerable variation of physical activity across EPIC centres. This variation was especially evident for recreational activities in both men and women. PMID- 12639226 TI - Consumption of vegetables, fruit and other plant foods in the European Prospective Investigation into Cancer and Nutrition (EPIC) cohorts from 10 European countries. AB - OBJECTIVE: To describe and compare the consumption of the main groups and sub groups of vegetables and fruits (V&F) in men and women from the centres participating in the European Prospective Investigation into Cancer and Nutrition (EPIC). DESIGN: Cross-sectional analysis. Dietary intake was assessed by means of a 24-hour dietary recall using computerised interview software and standardised procedures. Crude and adjusted means were computed for the main groups and sub groups of V&F by centre, separately for men and women. Adjusted means by season, day of the week and age were estimated using weights and covariance analysis. SETTING: Twenty-seven centres in 10 European countries participating in the EPIC project. SUBJECTS: In total, 35 955 subjects (13 031 men and 22 924 women), aged 35-74 years, randomly selected from each EPIC cohort. RESULTS: The centres from southern countries had the highest consumption of V&F, while the lowest intake was seen in The Netherlands and Scandinavia for both genders. These differences were more evident for fruits, particularly citrus. However, slightly different patterns arose for some sub-groups of vegetables, such as root vegetables and cabbage. Adjustment for body mass index, physical activity, smoking habits and education did not substantially modify the mean intakes of vegetables and fruits. CONCLUSIONS: Total vegetable and fruit intake follows a south-north gradient in both genders, whereas for several sub-groups of vegetables a different geographic distribution exists. Differences in mean intake of V&F by centre were not explained by lifestyle factors associated with V&F intake. PMID- 12639227 TI - Food sources of carbohydrates in a European cohort of adults. AB - OBJECTIVE: To describe the average consumption of carbohydrate-providing food groups among study centres of the European Prospective Investigation into Cancer and Nutrition (EPIC). METHODS: Of the 27 redefined EPIC study centres, 19 contributed subjects of both genders and eight centres female participants only (men, women, after exclusion of subjects under 35 and over 74 years of age from the original 36 900 total). Dietary data were obtained using the 24-hour recall methodology using the EPIC-SOFT software. The major sources of dietary carbohydrate were identified, and 16 food groups were examined. RESULTS: The 10 food groups contributing most carbohydrate were bread; fruit; milk and milk products; sweet buns, cakes and pies; potato; sugar and jam; pasta and rice; vegetables and legumes; crispbread; and fruit and vegetable juices. Consumption of fruits as well as vegetables and legumes was higher in southern compared with northern centres, while soft drinks consumption was higher in the north. Italian centres had high pasta and rice consumption, but breakfast cereal, potato, and sweet buns, cakes and pies were higher in northern centres. In Sweden, lower bread consumption was balanced with a higher consumption of crispbread, and with sweet buns, cakes and pies. Overall, men consumed higher amounts of vegetables and legumes, bread, soft drinks, potatoes, pasta and rice, breakfast cereal and sugar and jam than women, but fruit consumption appeared more frequent in women. CONCLUSION: The study supports the established idea that carbohydrate-rich foods chosen in northern Europe are different from those in the Mediterranean region. When comparing and interpreting diet-disease relationships across populations, researchers need to consider all types of foods. PMID- 12639228 TI - Soy product consumption in 10 European countries: the European Prospective Investigation into Cancer and Nutrition (EPIC) study. AB - OBJECTIVE: The aim of this study was to describe the variation of soy product intake in 10 European countries by using a standardised reference dietary method. A subsidiary aim was to characterise the pattern of soy consumption among a sub group of participants with a habitual health-conscious lifestyle (HHL), i.e. non meat eaters who are fish eaters, vegetarians and vegans. DESIGN: A 24-hour dietary recall interview (24-HDR) was conducted among a sample (5-12%) of all cohorts in the European Prospective Investigation into Cancer and Nutrition (EPIC). Study participants totalled 35 955 after exclusion of subjects younger than 35 or older than 74 years of age. Soy products were subdivided into seven sub-groups by similarity. Distribution of consumption and crude and adjusted means of intake were computed per soy product group across countries. Intake of soy products was also investigated among participants with an HHL. RESULTS: In total, 195 men and 486 women reported consuming soy products in the 24-HDR interview. Although soy product intake was generally low across all countries, the highest intake level was observed in the UK, due to over-sampling of a large number of participants with an HHL. The most frequently consumed soy foods were dairy substitutes in the UK and France and beans and sprouts among mid-European countries. For both genders, the sub-group of soy dairy substitutes was consumed in the highest quantities (1.2 g day-1 for men; 1.9 g day-1 for women). Participants with an HHL differed substantially from others with regard to demographic, anthropometric and nutritional factors. They consumed higher quantities of almost all soy product groups. CONCLUSIONS: Consumption of soy products is low in centres in Western Europe. Soy dairy substitutes are most frequently consumed. Participants with an HHL form a distinct sub-group with higher consumptions of fruit, vegetables, legumes, cereals and soy products compared with the other participants. PMID- 12639229 TI - Consumption of added fats and oils in the European Prospective Investigation into Cancer and Nutrition (EPIC) centres across 10 European countries as assessed by 24-hour dietary recalls. AB - OBJECTIVE: To evaluate the consumption of added fats and oils across the European centres and countries participating in the European Prospective Investigation into Cancer and Nutrition (EPIC). DESIGN AND SETTING: 24-Hour dietary recalls were collected by means of standardised computer-guided interviews in 27 redefined EPIC centres across 10 European countries. SUBJECTS: From an initial number of 36 900 subjects, single dietary recalls from 22 924 women and 13 031 men in the age range of 35-74 years were included. RESULTS: Mean daily intake of added fats and oils varied between 16.2 g (Varese, Italy) and 41.1 g (Malmo, Sweden) in women and between 24.7 g (Ragusa, Italy) and 66.0 g (Potsdam, Germany) in men. Total mean lipid intake by consumption of added fats and oils, including those used for sauce preparation, ranged between 18.3 (Norway) and 37.2 g day-1 (Greece) in women and 28.4 (Heidelberg, Germany) and 51.2 g day-1 (Greece) in men. The Mediterranean EPIC centres with high olive oil consumption combined with low animal fat intake contrasted with the central and northern European centres where fewer vegetable oils, more animal fats and a high proportion of margarine were consumed. The consumption of added fats and oils of animal origin was highest in the German EPIC centres, followed by the French. The contribution of added fats and oils to total energy intake ranged from 8% in Norway to 22% in Greece. CONCLUSIONS: The results demonstrate a high variation in dietary intake of added fats and oils in EPIC, providing a good opportunity to elucidate the role of dietary fats in cancer aetiology. PMID- 12639230 TI - Meat consumption in the European Prospective Investigation into Cancer and Nutrition (EPIC) cohorts: results from 24-hour dietary recalls. AB - OBJECTIVE: To evaluate meat intake patterns in the European Prospective Investigation into Cancer and Nutrition (EPIC) cohorts. DESIGN AND SETTING: 24 Hour dietary recalls were assessed within the framework of a prospective cohort study in 27 centres across 10 European countries by means of standardised computer-assisted interviews. SUBJECTS: In total, 22 924 women and 13 031 men aged 35-74 years. RESULTS: Mean total meat intake was lowest in the 'health conscious' cohort in the UK (15 and 21 g day-1 in women and men, respectively) and highest in the north of Spain, especially in San Sebastian (124 and 234 g day 1, respectively). In the southern Spanish centres and in Naples (Italy), meat consumption was distinctly lower than in the north of these countries. Central and northern European centres/countries showed rather similar meat consumption patterns, except for the British and French cohorts. Differences in the intake of meat sub-groups (e.g. red meat, processed meat) across EPIC were even higher than found for total meat intake. With a few exceptions, the Mediterranean EPIC centres revealed a higher proportion of beef/veal and poultry and less pork or processed meat than observed in central or northern European centres. The highest sausage consumption was observed for the German EPIC participants, followed by the Norwegians, Swedish, Danish and Dutch. CONCLUSIONS: The results demonstrate distinct differences in meat consumption patterns between EPIC centres across Europe. This is an important prerequisite for obtaining further insight into the relationship between meat intake and the development of chronic diseases. PMID- 12639231 TI - Consumption of dairy products in the European Prospective Investigation into Cancer and Nutrition (EPIC) cohort: data from 35 955 24-hour dietary recalls in 10 European countries. AB - OBJECTIVES: To describe and compare the consumption of dairy products in cohorts included in the European Prospective Investigation into Cancer and Nutrition (EPIC). METHODS: Data from single 24-hour dietary recall interviews collected through a highly standardised computer-based program (EPIC-SOFT) in 27 redefined centres in 10 European countries between 1995 and 2000. From a total random sample of 36 900, 22 924 women and 13 031 men were selected after exclusion of subjects under 35 and over 74 years of age. RESULTS: A high total consumption of dairy products was reported in most of the centres in Spain and in the UK cohort sampled from the general population, as well as in the Dutch, Swedish and Danish centres. A somewhat low consumption was reported in the Greek centre and in some of the Italian centres (Ragusa and Turin). In all centres and for both sexes, milk constituted the dairy sub-group with the largest proportion (in grams) of total dairy consumption, followed by yoghurt and other fermented milk products, and cheese. Still, there was a wide range in the contributions of the different dairy sub-groups between centres. The Spanish and Nordic centres generally reported a high consumption of milk, the Swedish and Dutch centres reported a high consumption of yoghurt and other fermented milk products, whereas the highest consumption of cheese was reported in the French centres. CONCLUSION: The results demonstrate both quantitative and qualitative disparities in dairy product consumption among the EPIC centres. This offers a sound starting point for analyses of associations between dairy intake and chronic diseases such as cancer. PMID- 12639232 TI - Variability of fish consumption within the 10 European countries participating in the European Investigation into Cancer and Nutrition (EPIC) study. AB - OBJECTIVE: To describe and compare the consumption of total fish (marine foods) and the fish sub-groups - white fish, fatty fish, very fatty fish, fish products and crustacea, in participants from the European Investigation into Cancer and Nutrition (EPIC) study. DESIGN: Cross-sectional analysis of dietary intake using a computerised standardised 24-hour recall interview. Crude means, means and standard errors adjusted by age, season and day of the week were calculated, stratified by centre and gender. SETTING: Twenty-seven redefined centres in the 10 European countries participating in the EPIC study. SUBJECTS: In total, 35 955 subjects (13 031 men and 22 924 women), aged 35-74 years, selected from the main EPIC cohort. RESULTS: A six- to sevenfold variation in total fish consumption exists in women and men, between the lowest consumption in Germany and the highest in Spain. Overall, white fish represented 49% and 45% of the intake of total fish in women and men, respectively, with the greatest consumption in centres in Spain and Greece and the least in the German and Dutch centres. Consumption of fatty fish reflected that of total fish. However, the greatest intake of very fatty fish was in the coastal areas of northern Europe (Denmark, Sweden and Norway) and in Germany. Consumption of fish products was greater in northern than in southern Europe, with white fish products predominating in centres in France, Italy, Spain, The Netherlands and Norway. Intake of roe and roe products was low. The highest consumption of crustacea was found in the French, Spanish and Italian centres. The number of fish types consumed was greater in southern than in northern Europe. The greatest variability in consumption by day of the week was found in the countries with the lowest fish intake. CONCLUSIONS: Throughout Europe, substantial geographic variation exists in total fish intake, fish sub-groups and the number of types consumed. Day-to day variability in consumption is also high. PMID- 12639233 TI - Patterns of alcohol consumption in 10 European countries participating in the European Prospective Investigation into Cancer and Nutrition (EPIC) project. AB - OBJECTIVE: The aim of this study was to compare the quantities of alcohol and types of alcoholic beverages consumed, and the timing of consumption, in centres participating in the European Prospective Investigation into Cancer and Nutrition (EPIC). These centres, in 10 European countries, are characterised by widely differing drinking habits and frequencies of alcohol-related diseases. METHODS: We collected a single standardised 24-hour dietary recall per subject from a random sample of the EPIC cohort (36 900 persons initially and 35 955 after exclusion of subjects under 35 and over 74 years of age). This provided detailed information on the distribution of alcohol consumption during the day in relation to main meals, and was used to determine weekly consumption patterns. The crude and adjusted (by age, day of week and season) means of total ethanol consumption and consumption according to type of beverage were stratified by centre and sex. RESULTS: Sex was a strong determinant of drinking patterns in all 10 countries. The highest total alcohol consumption was observed in the Spanish centres (San Sebastian, 41.4 g day-1) for men and in Danish centres (Copenhagen, 20.9 g day-1) for women. The lowest total alcohol intake was in the Swedish centres (Umea, 10.2 g day-1) in men and in Greek women (3.4 g day-1). Among men, the main contributor to total alcohol intake was wine in Mediterranean countries and beer in the Dutch, German, Swedish and Danish centres. In most centres, the main source of alcohol for women was wine except for Murcia (Spain), where it was beer. Alcohol consumption, particularly by women, increased markedly during the weekend in nearly all centres. The German, Dutch, UK (general population) and Danish centres were characterised by the highest percentages of alcohol consumption outside mealtimes. CONCLUSIONS: The large variation in drinking patterns among the EPIC centres provides an opportunity to better understand the relationship between alcohol and alcohol-related diseases. PMID- 12639234 TI - Trends in self-reported past alcoholic beverage consumption and ethanol intake from 1950 to 1995 observed in eight European countries participating in the European Investigation into Cancer and Nutrition (EPIC). AB - OBJECTIVE: To describe the trends of self-reported past consumption of alcoholic beverages and ethanol intake from 1950 to 1995 within the European Prospective Investigation into Cancer and Nutrition (EPIC). DESIGN: Data on consumption of beer/cider, wine and liqueur/spirits were obtained retrospectively at age 20, 30 and 40 years to calculate average consumption and ethanol intake for the time periods 1950-1975 (at age 20), 1960-1985 (at age 30) and 1970-1995 (at age 40). Regression analysis was conducted with the time period data to assess trends in past alcoholic beverage consumption and ethanol intake with time. SETTING: The EPIC project. SUBJECTS: In total, 392 064 EPIC participants (275 249 women and 116 815 men) from 21 study centres in eight European countries. RESULTS: Generally, increases in beer/cider consumption were observed for most EPIC centres for 1950-1975, 1960-1985 and 1970-1995. Trends in wine consumption differed according to geographical location: downward trends with time were observed for men in southern European EPIC centres, upward trends for those in middle/northern European study centres. For women, similar but less pronounced trends were observed. Because wine consumption was the major contributor to ethanol intake for both men and women in most study centres, time trends for ethanol intake showed a similar geographical pattern to that of wine consumption. CONCLUSION: The different trends in alcoholic beverage consumption and ethanol intake suggest that information depicting lifetime history of ethanol intake should be included in analyses of the relationship between ethanol and chronic diseases, particularly in multi-centre studies such as EPIC. PMID- 12639235 TI - Diversity of dietary patterns observed in the European Prospective Investigation into Cancer and Nutrition (EPIC) project. AB - OBJECTIVE: To describe the diversity in dietary patterns existing across centres/regions participating in the European Prospective Investigation into Cancer and Nutrition (EPIC). DESIGN AND SETTING: Single 24-hour dietary recall measurements were obtained by means of standardised face-to-face interviews using the EPIC-SOFT software. These have been used to present a graphic multi dimensional comparison of the adjusted mean consumption of 22 food groups. SUBJECTS: In total, 35 955 men and women, aged 35-74 years, participating in the EPIC nested calibration study. RESULTS: Although wide differences were observed across centres, the countries participating in EPIC are characterised by specific dietary patterns. Overall, Italy and Greece have a dietary pattern characterised by plant foods (except potatoes) and a lower consumption of animal and processed foods, compared with the other EPIC countries. France and particularly Spain have more heterogeneous dietary patterns, with a relatively high consumption of both plant foods and animal products. Apart from characteristics specific to vegetarian groups, the UK 'health-conscious' group shares with the UK general population a relatively high consumption of tea, sauces, cakes, soft drinks (women), margarine and butter. In contrast, the diet in the Nordic countries, The Netherlands, Germany and the UK general population is relatively high in potatoes and animal, processed and sweetened/refined foods, with proportions varying across countries/centres. In these countries, consumption of vegetables and fruit is similar to, or below, the overall EPIC means, and is low for legumes and vegetable oils. Overall, dietary patterns were similar for men and women, although there were large gender differences for certain food groups. CONCLUSIONS: There are considerable differences in food group consumption and dietary patterns among the EPIC study populations. This large heterogeneity should be an advantage when investigating the relationship between diet and cancer and formulating new aetiological hypotheses related to dietary patterns and disease. PMID- 12639238 TI - Key events in the history of electron microscopy. PMID- 12639236 TI - Evaluation of under- and overreporting of energy intake in the 24-hour diet recalls in the European Prospective Investigation into Cancer and Nutrition (EPIC). AB - OBJECTIVE: To evaluate under- and overreporting and their determinants in the EPIC 24-hour diet recall (24-HDR) measurements collected in the European Prospective Investigation into Cancer and Nutrition (EPIC). DESIGN: Cross sectional analysis. 24-HDR measurements were obtained by means of a standardised computerised interview program (EPIC-SOFT). The ratio of reported energy intake (EI) to estimated basal metabolic rate (BMR) was used to ascertain the magnitude, impact and determinants of misreporting. Goldberg's cut-off points were used to identify participants with physiologically extreme low or high energy intake. At the aggregate level the value of 1.55 for physical activity level (PAL) was chosen as reference. At the individual level we used multivariate statistical techniques to identify factors that could explain EI/BMR variability. Analyses were performed by adjusting for weight, height, age at recall, special diet, smoking status, day of recall (weekday vs. weekend day) and physical activity. SETTING: Twenty-seven redefined centres in the 10 countries participating in the EPIC project. SUBJECTS: In total, 35 955 men and women, aged 35-74 years, participating in the nested EPIC calibration sub-studies. RESULTS: While overreporting has only a minor impact, the percentage of subjects identified as extreme underreporters was 13.8% and 10.3% in women and men, respectively. Mean EI/BMR values in men and women were 1.44 and 1.36 including all subjects, and 1.50 and 1.44 after exclusion of misreporters. After exclusion of misreporters, adjusted EI/BMR means were consistently less than 10% different from the expected value of 1.55 for PAL (except for women in Greece and in the UK), with overall differences equal to 4.0% and 7.4% for men and women, respectively. We modelled the probability of being an underreporter in association with several individual characteristics. After adjustment for age, height, special diet, smoking status, day of recall and physical activity at work, logistic regression analyses resulted in an odds ratio (OR) of being an underreporter for the highest vs. the lowest quartile of body mass index (BMI) of 3.52 (95% confidence interval (CI) 2.91-4.26) in men and 4.80 (95% CI 4.11-5.61) in women, indicating that overweight subjects are significantly more likely to underestimate energy intake than subjects in the bottom BMI category. Older people were less likely to underestimate energy intake: ORs were 0.58 (95% CI 0.45-0.77) and 0.74 (95% CI 0.63-0.88) for age (> or =65 years vs. <50 years). Special diet and day of the week showed strong effects. CONCLUSION: EI tends to be underestimated in the vast majority of the EPIC centres, although to varying degrees; at the aggregate level most centres were below the expected reference value of 1.55. Underreporting seems to be more prevalent among women than men in the EPIC calibration sample. The hypothesis that BMI (or weight) and age are causally related to underreporting seems to be confirmed in the present work. This introduces further complexity in the within-group (centre or country) and between-group calibration of dietary questionnaire measurements to deattenuate the diet-disease relationship. PMID- 12639239 TI - Monet's painting under the microscope. AB - An oil painting by Claude Monet, Port-Goulphar, Belle-Ile 1887 (collection of the Art Gallery of New South Wales), was examined to determine both the identity of the pigments used by the artist in this painting and his technique of mixing colors and laying paint on the canvas. The extremely complex construction of the painting was revealed by optical microscopy, scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDS), and X-ray mapping (XRM) analysis of cross sections of paint flakes excised from damaged regions of Port-Goulphar, Belle Ile. Nine different pigments were found on the painting. Many of the identified colors were modern pigments that became available only late in the 19th century as a result of scientific advances in pigment chemistry. Although similar colors were available in a natural mineral form, they lacked the vivid color of their manufactured counterparts. The use of these new synthetic metallic oxide colors by Monet accounts for the brilliance of his paintings. In addition, a separation between successive paint layers was observed in some areas of paint chip cross sections, indicating that oil-based paint was applied to paint that had dried, and consequently, Port-Goulphar, Belle-Ile was painted over a long period of time. This observation is contrary to the general perception of Monet's technique of painting freely and quickly. PMID- 12639240 TI - Cathodoluminescence efficiency dependence on excitation density in n-type gallium nitride. AB - Cathodoluminescence (CL) spectra from silicon doped and undoped wurtzite n-type GaN have been measured in a SEM under a wide range of electron beam excitation conditions, which include accelerating voltage, beam current, magnification, beam diameter, and specimen temperature. The CL intensity dependence on excitation density was analyzed using a power-law model (I CL proportional, variant J m ) for each of the observed CL bands in this material. The yellow luminescence band present in both silicon and undoped GaN exhibits a close to cube root (m = 0.33) dependence on electron beam excitation at both 77 K and 300 K. However, the blue (at 300 K) and donor-acceptor pair (at 77 K) emission peaks observed in undoped GaN follow power laws with exponents of m = 1 and m = 0.5, respectively. As expected from its excitonic character, the near band edge emission intensity depends linearly (m = 1) in silicon doped GaN and superlinearly (m = 1.2) in undoped GaN on the electron beam current. Results show that the intensities of the CL bands are highly dependent not only on the defect concentration but also on the electron-hole pair density and injection rate. Furthermore, the size of the focussed electron beam was found to have a considerable effect on the relative intensities of the CL emission peaks. Hence SEM parameters such as the objective lens aperture size, astigmatism, and the condenser lens setting must also be considered when assessing CL data based on intensity measurements from this material. PMID- 12639241 TI - Application of quantitative analytical electron microscopy to the mineral content of insect cuticle. AB - Quantification of calcium in the cuticle of the fly larva Exeretonevra angustifrons was undertaken at the micron scale using wavelength dispersive X-ray microanalysis, analytical standards, and a full matrix correction. Calcium and phosphorus were found to be present in the exoskeleton in a ratio that indicates amorphous calcium phosphate. This was confirmed through electron diffraction of the calcium-containing tissue. Due to the pragmatic difficulties of measuring light elements, it is not uncommon in the field of entomology to neglect the use of matrix corrections when performing microanalysis of bulk insect specimens. To determine, firstly, whether such a strategy affects the outcome and secondly, which matrix correction is preferable, phi-rho (z) and ZAF matrix corrections were contrasted with each other and without matrix correction. The best estimate of the mineral phase was found to be given by using the phi-rho (z) correction. When no correction was made, the ratio of Ca to P fell outside the range for amorphous calcium phosphate, possibly leading to flawed interpretation of the mineral form when used on its own. PMID- 12639242 TI - Charge-related problems associated with X-ray microanalysis in the variable pressure scanning electron microscope at low pressures. AB - In variable pressure scanning electron microscopy (VPSEM) the current data suggests that considerable caution is required in the interpretation of X-ray data from nonconductive samples, depending on the operating conditions. This article reviews some of the documented approaches and presents data that illustrate the nature and magnitude of the effects of charge above, on, and in the sample on the detected X-ray emissions from the sample and from elsewhere within the VPSEM specimen chamber. The collection of reliable and reproducible X ray data has been found to require relatively high specimen chamber gas pressures, at the upper end of or beyond the available pressures for most VPSEMs. It is also shown that sample characteristics, including composition, strongly influence local charge effects, which can significantly affect the primary electron landing energy and consequently the resultant emitted X-ray signal under low pressure environments. PMID- 12639243 TI - Human immunodeficiency virus type 1 subtype and other factors associated with extrapulmonary Cryptococcosis among patients in Thailand with AIDS. AB - Delineating factors associated with extrapulmonary cryptococcosis (EPC), a major disease burden among Thailand's AIDS patients, can clarify its pathogenesis and guide preventive interventions. From November 1993 through June 1996, enhanced surveillance of 2261 human immunodeficiency virus (HIV)-seropositive patients in a hospital near Bangkok showed EPC among 561 of 1553 AIDS patients (36.1%). Univariate analysis results were confirmed by multivariate analyses of data on 1259 patients. Logistic regression models identified factors significantly associated with EPC to be male sex (adjusted odds ratio [aOR], 2.0; 95% confidence interval [CI], 1.3-2.9), age <33 years (aOR, 1.5; 95% CI, 1.2-1.9), severe immunosuppression (aOR, 1.8; 95% CI, 1.3-2.6), not injecting drugs (aOR, 3.0; 95% CI, 1.7-5.5), and infection with HIV-1 circulating from CRF01_AE (formerly subtype E) versus subtype B (aOR, 2.3; 95% CI, 1.2-4.5). The association with CRF01_AE may result from undetermined markers of exposure or viral subtype effects on host immune responses. Better understanding of the epidemiology of EPC may reduce EPC incidence through targeted primary prevention and treatment. PMID- 12639245 TI - Thymic output during initial highly active antiretroviral therapy (HAART) and during HAART supplementation with interleukin 2 and/or with HIV type 1 immunogen (Remune). AB - The thymic output of patients receiving highly active antiretroviral therapy (HAART) was assessed by sjTREC (signal joint T cell receptor rearrangement excision circle) analysis to determine the thymic contribution to CD4(+) T cell reconstitution during initial therapy and during interleukin 2 (IL-2) and/or Remune supplementation of HAART. Levels of sjTRECs were observed to decline dramatically after the first 4 weeks of HAART and then increased without significant associated changes in CD4(+) T cell counts. HAART supplementation with IL-2 was observed to lead to rapid increases in CD4(+) T cells that were accompanied by sjTREC decreases. No notable changes in CD4(+) T cell counts and sjTRECs were seen in patients receiving HAART supplemented with Remune alone. The results indicate CD4(+) T cell maintenance during initial treatment of HIV-1 with HAART and early CD4(+) T cell reconstitution of patients receiving IL-2 with HAART is largely due to thymus-independent mechanisms, with the thymus making a limited contribution. PMID- 12639244 TI - In vivo antiviral activity of novel human immunodeficiency virus type 1 nucleocapsid p7 zinc finger inhibitors in a transgenic murine model. AB - Control of human immunodeficiency virus through the use of inexpensive chemotherapeutics, with minimal side effects and decreased potential for engendering resistant virus, is a long-term therapeutic goal. In principle, this goal can be accomplished if viral replication in reservoirs of chronically and latently infected cells is addressed. As a first step, we have developed novel antiviral compounds based on a 2-mercaptobenzamide thioester chemotype, including the pyridinioalkanoyl thioesters, which specifically target the zinc fingers of the human immunodeficiency virus nucleocapsid protein (NCp7). Using these compounds in a murine transgenic model, in which infectious human immunodeficiency virus is induced from an integrated provirus, we show inhibition of transgenic spleen cell p24 expression with potencies comparable to acute infection assays using human peripheral blood lymphocytes. More importantly, transgenic mice treated in vivo with two 2-mercaptobenzamide thioesters expressed significantly lower plasma p24, and splenocytes from these animals produced fewer infectious virions. Thus, these thioesters may provide an effective means for inhibiting the expression of human immunodeficiency virus from integrated viral reservoirs. PMID- 12639246 TI - Increased intrathecal release of soluble fractalkine in HIV-infected patients. AB - The CX(3)C chemokine fractalkine is suggested to play an important role in inflammatory brain diseases, for example, because of its chemotactic properties. To investigate the release of soluble fractalkine in HIV-induced brain diseases fractalkine levels were determined in cerebrospinal fluid (CSF) and serum samples of HIV-infected patients with (n = 10) and without (n = 23) HIV-induced CNS complications, using semiquantitative Western blot analysis. Fractalkine CSF levels were significantly elevated (p < 0.05) in HIV-infected patients with CNS diseases compared with those without, and compared with HIV-negative controls (n = 23). Fractalkine serum concentrations did not differ between the two groups of HIV-infected patients, but were significantly elevated (p < 0.05) in HIV-infected patients with CNS complications compared with HIV-negative controls. Levels of fractalkine did not correlate with the CSF and serum HIV load and other CSF parameters. In one patient with HIV-associated dementia and myelopathy CSF fractalkine levels decreased on initiation of antiretroviral therapy and subsequent clinical improvement. In conclusion, intrathecal fractalkine release was observed in the majority of patients with HIV infection. The highest levels of soluble fractalkine were detected in CSF (and serum) samples of patients with HIV-induced CNS disorders. These results suggest a dysregulation of brain soluble fractalkine release during HIV infection. PMID- 12639247 TI - Surface CD4 expression modulated by a cellular factor induced by HIV type 1 infection. AB - Human immunodeficiency virus type 1 (HIV-1) alters gene expression in infected cells, leading to cellular dysfunction. We uncovered a number of host cell genes that are modulated in both CD4(+) T cell lines and primary CD4(+) T lymphocytes infected with HIV-1, using high-density oligonucleotide probe microarray technology. We focused on one gene in particular, nuclear factor I-B2 (NFI-B2), because of its high level of expression. NFI-B2 is a member of the nuclear factor I family of nuclear proteins, which are known to be involved in viral and cellular transcription. To better understand the role of NFI-B2 during HIV-1 infection, we generated a Jurkat T cell line that constitutively expressed NFI B2. After infection with HIV-1, these cells produced fewer viruses because of a downregulation of surface CD4 expression. The surface expression of the coreceptor, CXCR4, remained unchanged. Furthermore, levels of CD4 mRNA were reduced in NFI-B2-producing cells, suggesting that expression of NFI-B2 impairs CD4 transcription. Modulation of NFI-B2 by HIV-1 may represent yet another mechanism by which HIV infection reduces cell surface expression of CD4. PMID- 12639248 TI - Primary African HIV clade A and D isolates: effective cross-clade neutralization with a quadruple combination of human monoclonal antibodies raised against clade B. AB - We investigated the ability of several human neutralizing monoclonal antibodies (nmAbs), originally raised against human immunodeficiency virus (HIV) clade B isolates, to neutralize primary clade A and D isolates as single agents and in combinations. All four primary HIV clade A isolates and five primary HIV clade D isolates tested were neutralized >99% by the quadruple combination of nmAbs IgG1b12, 2G12, 2F5, and 4E10. These mAbs recognize conserved epitopes on HIV-1 envelope (Env), resulting in strong cross-clade neutralization. Previously, we showed synergistic neutralization of primary HIV-1 clade C isolates in vitro by the same nMAb combination. We and others also showed neutralization of primary HIV clade B strains. Together, our data show that the quadruple combination of mAbs effectively neutralized primary HIV clade A, B, C, and D isolates. PMID- 12639250 TI - Analytical procedure for the determination of chlorobenzenes in sediments. AB - This study presents the procedure for the determination of chlorobenzenes in sediment. It consists of solvent extraction (shaking overnight), extract clean-up with the use of a homemade glass column packed with activated silica gel and freshly activated copper, and slow solvent evaporation to a volume of 0.3 mL. Two microliter extract portions are analyzed by means of gas chromatography with an Rtx-624 capillary column (60 m x 0.32 mm, d(f) = 1.8 microm) coupled with mass spectrometry (in selected ion-monitoring mode). Deuterated 1,2-dibromobenzene is used as the recovery standard. The recovery of this method for all chlorobenzenes is high (ranging from 78% to 107%) with the exception of monochlorobenzene, which is 58%. The method is also characterized by good precision, which is commonly accepted in the analysis of trace organic pollution. PMID- 12639249 TI - Characterization and selection of HIV-1 subtype C isolates for use in vaccine development. AB - HIV-1 genetic diversity among circulating strains presents a major challenge for HIV-1 vaccine development, particularly for developing countries where less sequence information is available. To identify representative viruses for inclusion in candidate vaccines targeted for South Africa, we applied an efficient sequence survey strategy to samples from recently and chronically infected persons residing in potential vaccine trial sites. All 111 sequences were subtype C, including 30 partial gag, 26 partial pol, 27 V2-V3 env, and 28 V5 partial gp41 sequences. Of the 10 viruses cultured from recently infected individuals, 9 were R5 and 1 was R5X4. Two isolates, Du151 and Du422, collected within 2 months of infection, were selected as vaccine strains on the basis of their amino acid similarity to a derived South African consensus sequence The selection of recently transmitted R5 isolates for vaccine design may provide an advantage in a subtype C R5-dominant epidemic. The full-length Du422 gag and Du151 pol and env genes were cloned into the Venezuelan equine encephalitis (VEE) replicon particle (VRP) expression system. Du422 Gag protein expressed from the VRP accumulated to a high level and was immunogenic as demonstrated by cytotoxic T lymphocyte responses in mice vaccinated with gag-VRPs. Optimization of codon use for VRP expression in human cells did not enhance expression of the gag gene. The cloned Du151 env gene encoded a functional protein as demonstrated by fusion of VRP-infected cells with cells expressing CD4 and CCR5. Genes identified in this study have been incorporated into the VEE VRP candidate vaccines targeted for clinical trial in South Africa. PMID- 12639251 TI - Rapid analysis of phentolamine by high-performance liquid chromatography. AB - A rapid liquid chromatographic method is validated for the quantitative analysis of phentolamine. Phentolamine exists in three forms for this investigation: as a mesylate salt, hydrochloride salt, and free base. In solution, phentolamine dissociates from its salt and is chromatographed as free phentolamine. This validation confirms the analysis of each form, which is simply based upon molar mass differences encountered in weighing. As such, both the United States Pharmacopeia hydrochloride and mesylate standards are used throughout this validation to demonstrate this equivalency. The validation demonstrates that this method may be used to quantitate phentolamine, regardless of its salt form. PMID- 12639252 TI - Sensitive analysis of alkyl alcohols as decomposition products of alkyl nitrites in human whole blood and urine by headspace capillary GC with cryogenic oven trapping. AB - The abuse of alkyl nitrites is becoming a serious social problem worldwide. In this report, a simple and sensitive method is presented for the determination of n-butyl alcohol, isobutyl alcohol, and isoamyl alcohol as decomposition products of alkyl nitrites in human whole blood and urine samples using capillary gas chromatography (GC) with cryogenic oven trapping. After heating a whole blood or urine sample containing each alkyl alcohol and t-butyl alcohol [the internal standard (IS)] in a 7-mL vial at 55 degrees C for 15 min, 5 mL of the headspace vapor is drawn into a gas-tight syringe and injected into a GC inlet port. The vapor is introduced into an Rtx-BAC2 medium-bore capillary column in the splitless mode at 0 degrees C oven temperature in order to trap the entire analytes, and then the oven temperature is programmed up to 240 degrees C for the GC measurements by flame ionization detection. These conditions give sharp peaks for each compound and the IS and low background noise for whole blood or urine samples. The detection limits of the analytes are 10 ng/mL for whole blood and 5 ng/mL for urine. Linearity and precision are also tested to confirm the reliability of this method. Isobutyl alcohol and methemoglobin could be determined from the whole blood samples of three male volunteers who had sniffed isobutyl nitrite. PMID- 12639253 TI - HPLC-electrospray ionization-MS-MS analysis of Cephalotaxus harringtonia leaves and enhancement of the extraction efficiency of alkaloids therein by SFE. AB - A high-performance liquid chromatographic method is developed for use in the electrospray mass spectrometric (MS)-MS analysis of alkaloids contained in Cephalotaxus harringtonia leaves. Nine alkaloids having ester groups can be separated and detected with good sensitivity. The MS and MS-MS spectra obtained provides information on their chemical structures. Supercritical fluid extraction is also applied in order to improve the extraction efficiency of Cephalotaxus alkaloids such as cephalotaxine, harringtonine, homoharringtonine, and isoharringtonine. When carbon dioxide-methanol-water (80:18:2, v/v) is used the extraction yield is found to be higher than that using the other supercritical solvents evaluated and conventional organic solvent extraction. PMID- 12639254 TI - The USACHPPM gas chromatographic procedures for the analysis of waters and soils for energetics and related compounds. AB - The procedures currently used by the U.S. Army Center for Health Promotion and Preventive Medicine (USACHPPM) for the analysis of energetics and related compounds in water and soils are presented. These procedures are based on the use of isoamyl acetate to extract the analytes of interest from their environmental matrices with subsequent analysis using gas chromatography with electron capture detection. The suite of compounds included are those that have been of environmental significance for years (such as 2,4,6-trinitrotoluene, hexahydro 1,3,5-trinitro-1,3,5-triazine, and dinitrotoluenes) and are the subject of several U.S. Environmental Protection Agency SW-846 methods. The procedures presented in this study are the product of years of development and refinement of methods used for the analysis of many real-world samples by the USACHPPM explosives analysis laboratory. The development, performance, advantages, and details of these procedures are described. The extension of these methods to the analysis of other media is also briefly discussed. PMID- 12639256 TI - Determination of flavones in Crataegus pinnatifida by capillary zone electrophoresis. AB - A capillary electrophoretic method for the separation of four flavones in Crataegus pinnatifida is developed. The four flavones in Crataegus pinnatifida are separated on baseline within 15 min using 50mM borax buffer containing 15% acetonitrile and adjusted to pH 8.15 with phosphoric acid. The detection limits of vitexin-2"-rhamnoside, hyperside, rutin, and vitexin are 0.35, 0.30, 0.40 and, 0.29 microg/mL, respectively. The recovery of these flavones is as follows: vitexn-2"-rhamnoside 96.8%, hyperside 99.9%, rutin 97.1%, and vitexin 97.8%. The results are in accordance with those obtained in the high-performance liquid chromatography system. The content of flavones is higher in Crataegus pinnatifida leaves than in its fruits, and hyperside is not detected in either Crataegus pinnatifida fruits or flowers. PMID- 12639255 TI - Performance analysis of a rapid HPLC determination with the solvent demixing extraction of HIV antiproteases and efavirenz in plasma. AB - A rapid and simple high-performance liquid chromatographic method without internal standardization is evaluated for the drug-level monitoring of most marketed antiproteases and efavirenz. Following plasma deproteinization with acetonitrile, the analytes are extracted into the solvent while it is demixed by the addition of a saturating amount of neutral salt. The organic supernatant is diluted by half with water up to the polarity of the mobile phase before being injected. The isocratic mobile phase is unbuffered water-acetonitrile (52:48), and the stationary phase is LiChrospher 100 RP-8 (5 microm). Analytes are eluted between 4 min (amprenavir and indinavir) and 20 min (nelfinavir). A spreadsheet program including analysis of variance (ANOVA) and regression is used for both the overall validation of milligrams-per-liter determinations and the performance evaluation of analytical steps from chromatographic raw data. Extraction shows acceptable 5% repeatability and nearly 100% recovery, although it is somewhat concentration-dependent. The calibration function is better fitted by bilogarithmic than arithmetical regression, and the ANOVA of raw data is found quite predictive of the quality of the final determinations. PMID- 12639257 TI - Screening of pesticides via solid-phase extraction and gas chromatography. AB - This work describes an analytical method for the screening of pesticide residues in water. The developed method identifies and quantitates a variety of pesticides in water samples such as organochlorine, organophosphate, chloronitrile, phthalimide, dicarboximide, and triazine. The analytical technique employed for the extraction and cleanup step involves solid-phase extraction with C-18 cartridges. The identification and determination of concentration is carried out with gas chromatography. Two columns of different polarity and two specific detectors are used. An electron capture detector is used for organochrolorine, and a thermoinic detector is used for organonitrogen and organophosphate. The good resolution achieved with the temperature program and the combination of columns is shown in the chromatograms obtained. The limits of detection, percent recovery, and their respective standard deviations are determined for each product. According to the results obtained, this method permits monitoring of the pesticides mentioned, many of which are moderately to highly toxic to aquatic organisms. PMID- 12639258 TI - The optimization of HPLC-UV conditions for use with FTIR detection in the analysis of B vitamins. AB - The water-soluble vitamins thiamine (B(1)), riboflavin (B(2)), pantothenic acid (B(5)), and pyridoxine (B(6)) are separated by high-performance liquid chromatography. The mobile phase, column temperature, and flow rate are optimized so that the chromatograph can be used with a Fourier-transform infrared (FTIR) detector. Reproducibility, linearity, and detection limits are evaluated for method validation. Finally, this method is successfully transferred to liquid chromatography-FTIR with a standard mixture. PMID- 12639259 TI - Improvements to EPA method 531.1 for the analysis of carbamates that resulted in the development of U.S. EPA Method 531.2. AB - This project is undertaken to fully optimize the U.S. Environmental Protection Agency Method 531.1 post-column chemistries and to incorporate recent advances in liquid chromatographic separation, post-column derivatization, and detection techniques. Sample preservation and storage stability studies establish citric acid as a suitable replacement for the caustic monochloroacetic acid in the current method and confirm its antimicrobial effectiveness. Performance of an alternate set of commercially available post-column reagents is also investigated. This research has resulted in the publication of Method 531.2, a high-performance liquid chromatographic direct injection method for the analysis of N-methylcarbamoyloximes and N-methylcarbamates using post-column derivatization and fluorescence detection. PMID- 12639260 TI - The relationship between psychoanalysis and schizophrenia. AB - In this article, the author considers psychoanalysts' current attitudes towards schizophrenia. After early optimism of a psychoanalytic approach, interest has waned, other than in the field of first-onset psychosis. This was because of poor outcome figures and regarding schizophrenia as now having a biological, rather than psychological, base. The author argues that there is a paradox, because only psychoanalysis offers a framework for relating to psychotic patients in a way that helps them to make sense of their experiences. A framework is described, with clinical examples, to illustrate the application of analytic thinking to patients with schizophrenia. Psychoanalysis needs to revitalize its attitude to psychosis, as it has a significant contribution to make within general psychiatry, not least in the training of the next generation of psychiatrists. PMID- 12639261 TI - On not being able to dream. AB - In this paper, the author explores the phenomenon of not being able to dream (as opposed to not being able to remember one's dreams) from three different vantage points. First, from the point of view of psychoanalytic theory, he discusses Bion's idea that the work of dreaming creates the conscious and unconscious mind (and not the other way around). A person who cannot dream is unable to generate differentiable conscious and unconscious experience and, consequently, lives in a psychic state in which he is unable to differentiate waking from sleeping, dreaming from perceiving. The author then approaches the problem of the inability to dream from the perspective achieved by a literary work. He discusses a Borges fiction that creates, in a singularly artful way, the experience of not being able to dream. Finally, the author utilises the vantage point of a detailed account of a clinical experience to explore what it means not to be able to dream. He describes an initial state characterised by the patient's proliferation of unutilisable 'psychic noise' which, over a period of years, led to the analyst's experiencing 'reverie-deprivation' and brief periods of countertransference psychosis. Two analytic sessions are presented and discussed in which psychological work was done that contributed to an enhanced capacity on the part of both patient and analyst for genuine dreaming - both in sleep and in analytic reverie states. PMID- 12639262 TI - On psychoanalytic figures as transference objects. AB - One of the tasks that analysts and therapists face at a certain stage in their career is how to develop a way of psychoanalytic thinking and practising of their own. To do this involves modifying or overcoming the transferences established during their training or early career. These transferences are to one's teachers or training analyst, investing them with authority and infallibility, and to received theory, which is treated as though it were dogma. The need to free oneself from such transferences has been discussed in the literature. There is, however, another kind of transference that the developing therapist also needs to resolve, which has received little attention. This is the transference made on to a key figure in the psychoanalytic tradition. Such a psychoanalytic figure will be seen as the originator of or embodiment of those theoretical ideas to which one becomes attached, and/or as standing behind one's training analyst or seminal teachers who become a representative of that figure. The value of an investigation of one's relationship to a psychoanalytic figure is that it is an excellent medium for revealing one's transference, as the figure in question is not a real person but only exists through his/her writings. The body of the paper consists of an extended example of such an analysis, that of my own transference on to the figure of Winnicott. In this example I illustrate how my evaluation of Winnicott's ideas changed from seeing them as providing answers to all my clinical questions to no longer satisfying me in some areas of my work. This change in my relationship to Winnicott's theory went hand in hand with a modification in my transference on to the figure of Winnicott, from seeing him as endowed with authority and goodness to an appreciation of him as a still sustaining figure but now with limits and flaws. In the final part of the paper several questions arising out of my analysis are posed. Can the pull of writing such an account in terms of dramatic rupture rather than gradual and partial change be avoided? Should my account be regarded purely as a form of self analysis or does it have anything to say about Winnicott himself and his theory? And do some psychoanalytic figures attract more intense or sticky transferences than others? PMID- 12639263 TI - Salieri's dilemma: a counterpoint between envy and appreciation. AB - Taking as its starting point Klein's concepts of 1957, this paper stresses certain characteristics of envy that have not yet been taken into account in psychoanalysis. Klein described the conflict between envy and gratitude as inherent in the human being; she suggested an interesting link between envy and admiration, but never developed this idea in her theory. The hypothesis that we put forward in this paper is the existence of a particular intolerance-related to envy-of one's own capacity to recognise the valuable aspects of the object. This situation leads to the paradox that the same faculty that allows the patient to appreciate the good qualities of the object is at the same time the source of unbearable pain. The capacity to recognise the good aspects of the object-whether they are perceived or attributed by projective identification-cannot be accepted as a valuable aspect of the self and is taken as a proof of the self 's unworthiness. We think that the description of this intolerance towards admirative appreciation introduces a change in the understanding of idealization simply as a defence against envy and enriches the conflict expressed in the immediacy of the transference - countertransference. PMID- 12639264 TI - Preconscious defence analysis, memory and structural change. AB - Beginning with the ways in which the use of the couch lends 'depth to the surface' (Erikson, 1954), I explore the topography of the inter- and intrasubjective psychoanalytic situation and process. I suggest that defences are not by definition unconscious but rather can be observed operating at conscious and preconscious levels, particularly under these conditions. A focus on preconscious disavowal provides a window on what has become unconscious repression. As a result of eliciting and then verbalising the operation of such defences with regard to anxieties in the here-and-now transference, declarative memories of increasingly specific childhood fantasies and events begin to hold sway over unmanageable procedural remnants from the analysand's past. With this may even come the possibility of neuronal regeneration, the more generalisable enhancement of declarative and symbolic functions and the sense of identity with which these are associated. Herein may lie one enduring therapeutic effect of the 'talking cure' - putting feelings into words - as one among a variety of psychotherapeutic modalities. PMID- 12639265 TI - Candidate progression in analytic institutes: a multi-center study. AB - This study documents the criteria for graduation and for awarding credit for training cases used by thirteen institutes of the American Psychoanalytic Association (APsA). At the time of this study, these thirteen institutes were training 63% of all candidates enrolled at institutes of the APsA. Questionnaires were sent to the progression committee chairperson (PCC), a junior candidate and a senior candidate, and a recent graduate at each institute. Each participant also had a follow-up semi-structured telephone interview. Return rate was 100%. 92% per cent of the institutes reported that in order to receive credit for cases, the patient needed to be in treatment for some minimum amount of time (average 18.3 months). Many PCCs were skeptical about writing down the criteria for credit, stating that this made candidates overly focused on the time requirement. 85% per cent of PCCs felt that graduation from the institute was based primarily on obtaining credit for cases. Few required a terminated case and only one felt that classroom work was an important factor. 47% per cent of candidates felt that the need to keep patients in treatment to receive credit for cases affected their treatment of patients and only 46% of candidates could state the minimum time requirement used by their institute. These results indicate that graduation from institutes of the APsA depends primarily on receiving credit for cases, that case length is an important criterion for awarding credit and that this method of evaluating candidates may not be the best way to support educational objectives. PMID- 12639266 TI - A framework and two sketches concerning psychoanalytical training and practice. AB - The author discusses the mode of knowledge production in psychoanalysis based on a reflection on the psychoanalytical education and its relationship to clinical practice. She points out that there is a risk in a form of clinical activity found in our education, which, under the fascination of the analyst's power of operating in the metaphorical domain of words, loses sight of the material dimension of the clinical action. In other words, this form of clinical activity loses sight of the meeting with another human being, of the repertoire of theories and experiences that informs this action and the patient's and the analyst's concrete life situation. The author highlights the role of writing as a privileged way of dealing with the material and immaterial facts that constitute the clinical action and reflects on some of the forces that structure nowadays the reception of knowledge production inside the psychoanalytical field. She uses the notion of 'minor literature', by Franz Kafka, to express the possibility that a live circuit of writings exchanged among psychoanalysts can offer to an interchange of experiences and ideas that is the live expression of the history of the psychoanalytical groups. A clinical session is presented in order to promote considerations about the psychoanalytical education, theory and practice. PMID- 12639267 TI - Something more than the 'something more than interpretation' is needed: a comment on the paper by the Process of Change Study Group. AB - A critique is offered of the paper on non-interpretive mechanisms in psychoanalytic therapy by the Process of Change Study Group (Stern et al., 1998a). The attention paid to procedural learning and to the need to attend to implicit relational knowledge is welcomed but a number of the group's assumptions are questioned. In particular, it is argued that the differences between mother infant and therapist-patient relationships are inadequately considered, that the polarisation of non-symbolic and verbal memory is unhelpful and that the separation of the therapy relationship into three strands is problematic. The assumption that pursuing the goal of intersubjective understanding is in itself therapeutic is questioned. It is suggested that the recognition of the importance of implicit relational knowledge calls for a wider reappraisal of important aspects of theory and practice and that direct means of making such knowledge explicit should be considered. PMID- 12639268 TI - Reader and story, viewer and film: on transference and interpretation. AB - Two sides in Freud's attitude towards literature and art are presented: Freud the sensitive listener, whose interest in art is a potential springboard for a rich interdisciplinary dialogue; and Freud the conquistador, whose wish for power in 'invaded' territories is related to troublesome aspects of 'pathography' and 'applied analysis'. The unique contribution of psychoanalysis may not be discovering objectively the true unconscious content of works of art, but rather enriching the exploration of the potential transitional space evolving between artist, work of art and readers or viewers, enhancing our sensitivity to multiple meanings and complex emotional influences of art. This requires exploring our own subjective experiences of art, which may be described as transferences (when art is mostly perceived as a source of insight) or countertransferences (when artists and their work are basically experienced as troubled patients). Transference (broadly defined) and interpretation tend to intermingle, both in the clinical analytic encounter, and in any reading/viewing of art, be it by laymen, analysts or other scholars. Several examples from the psychoanalytic study of literature and film are given, and three pairs of contrasting interpretations are studied, concerning Kafka's The metamorphosis, Minghella's The English Patient and Polanski's Chinatown. PMID- 12639269 TI - Psychoanalytic interpretations: word-music and translation. AB - A perspective on psychoanalytic interpretations as a special case of artistic translations (i.e. translations of both content and formal aspects of discourse) is proposed. Mutative interpretations are seen as creative endeavours that always presuppose a prior text, which is itself already a translation. Freud's main uses of the metaphor of translation as an intrapsychic as well as an intersubjective phenomena, their resonance among later psychoanalytic thinkers and their convergence with classic theories of translation are presented. A perspective on psychoanalytic interpretations as an evolving self-other dialect is developed within the framework of Borges's theory of translation and Bakhtin's concept of exotopy. This framework questions the possibility of an original source, proposes the translator's inevitable implication in translation and underscores intersubjectivity as a necessary way towards the translation of absent meanings. The work of translation also highlights the relevance of space and time contexts, as well as the necessary role of a different other for the interpretation of unconscious processes. Differences between theories of translation reiterate contemporary psychoanalytic controversies about the preeminence of the analysand's subjectivity and the intersubjective approach to treatment. Borges's 'The Aleph' and an excerpt of Dora's case offer paradigmatic demonstrations of the limits and possibilities of understanding through translation. PMID- 12639270 TI - Reassessment of psychoanalytical education: controversies and changes. PMID- 12639271 TI - Panel on 'acting out and/or enactment'. PMID- 12639272 TI - Perception. AB - The article is an overview of some central philosophical problems associated with perception. It discusses what distinguishes perception from other sensory capacities and from conception. It discusses anti-individualism, a view according to which the nature of a perceptual state is dependent not just causally but for its identity or 'essence' on relations to a normal environment in which systems containing that state were formed. It discusses different views about epistemic warrant. By emphasising the deep ways in which human and animal perceptual systems, especially visual systems, are similar, it criticises a dominant view of the last century, in both philosophy and large parts of psychology, according to which a range of sophisticated supplementary abilities have to be learned before a child can perceive objective features of the physical world. PMID- 12639273 TI - The castaway self: a psychoanalytic reading of Castaway (2000). PMID- 12639274 TI - Genetics and educational psychology. AB - BACKGROUND: Molecular genetics, one of the most energetic and exciting areas of science, is slowly but surely coming to educational psychology. AIMS: We review recent molecular genetic research on learning disabilities as a sign of things to come in educational psychology. We also consider some misconceptions about genetics that have slowed the acceptance of genetics in educational psychology. SAMPLES: Diverse samples of children with learning disabilities have been studied, primarily in the UK and the USA. METHODS: Linkage analysis can detect genes that have large effects on learning disabilities. Association analysis can detect genes of much smaller effect size, which is important because common disorders such as learning disabilities are likely to be influenced by many genes as well as by many environmental factors. RESULTS: For reading disability, replicated linkages have been identified on chromosomes 6, 15 and 18. A gene responsible for a rare type of language impairment has recently been identified. For common language impairment, linkages on chromosomes 16 and 19 have recently been reported. More than 200 genetic disorders, most extremely rare, include mental retardation among their symptoms, and chromosomal abnormalities are a major cause of mental retardation. CONCLUSIONS: Although finding specific genes associated with learning disabilities is unlikely to have much of a direct application for teachers in the classroom, such findings will have far-reaching implications for diagnosis, treatment and prevention of learning disabilities and for research in educational psychology. Educational psychology has been slower to accept evidence for the importance of genetics than other areas of psychology in part because of misconceptions about what it means to say that genetics is important for common complex disorders such as learning disabilities. PMID- 12639275 TI - Class size, pupil attentiveness and peer relations. AB - BACKGROUND: Despite a long-running debate over the effects of class size differences on educational performance there is little evidence on the classroom processes that might be involved. AIMS: The effects of class size differences are examined in relation to social and behavioural adjustment to school, in terms of two dimensions: attentiveness and peer relations. It was predicted that as class size increased there would be more inattentiveness in class and more signs of social difficulties between children in the form of more rejection, asocial, anxious and aggressive behaviour, and less prosocial behaviour. SAMPLES: Data came from a large-scale longitudinal study of children over KS1 (4-7 years). The observation study was based on a subsample of 235 children in 21 small (average 19 children) and 18 large (average 33 children) reception classes (aged 5 years). The PBR sample involved over 5,000. METHODS: There were two complementary methods of data collection: first, a systematic observation study of pre-selected target children in terms of three 'social modes' - when with their teachers, other children and when not interacting - and in terms of work, procedural, social and off-task activities; and, second, a teacher administered Pupil Behaviour Rating (PBR) scale comprising over 50 items rated on a 3-point scale grouped into six 'factors': hyperactive/distractible, aggressive, anxious/ fearful, prosocial, asocial, and excluded. RESULTS: Observations showed that children in large classes were more likely to show off-task behaviour of all kinds, and more likely to interact with their peers in terms of off-task behaviour, social, and also on task behaviours. Connections between class size and PBR factors were not strong. There was no support for the view that peer relations are better in smaller classes; indeed, there was a slight tendency for worse peer relations, in terms of aggression, asocial and excluded, in the smallest classes. CONCLUSIONS: There was confirmation that children in large classes are more distracted from work and more often off task. The unexpected result, based on teacher ratings, that small classes may lead to less social and more aggressive relations between children is discussed, along with implications for teachers of a tendency for more peer related contacts in large classes. PMID- 12639276 TI - Tape-assisted reciprocal teaching: cognitive bootstrapping for poor decoders. AB - BACKGROUND: Students who have limited skills in decoding and comprehension and who lack motivation to read present difficulties for practitioners. Difficulties may be compounded when these students lack access to age-appropriate and interesting text and have lost the notion of reading as a process of obtaining meaning from print. AIMS: This research examined the effects of a modified reciprocal teaching intervention for readers with poor decoding skills and poor comprehension. Tape-assisted reciprocal teaching was used to help students with poor decoding skills develop cognitive and metacognitive strategies and improve their comprehension of high interest expository texts. METHODS: Two single subject research design studies involving four groups of students were conducted. Study I involved one experimental group and Study II was a multiple baseline design involving three experimental groups. SAMPLE: Each experimental group comprised a heterogeneous mix of six students, three with poor decoding skills and three with adequate decoding skills, all of whom showed poor comprehension. RESULTS: As a result of the tape-assisted reciprocal teaching, the poor decoders demonstrated improved application of cognitive and metacognitive strategies and improved comprehension. These improvements were shown on both researcher developed and standardised tests as well as on maintenance and transfer measures. The students with adequate decoding skills also showed improvements in comprehension. CONCLUSIONS: The success of the intervention for poor decoders suggests that tape assisted reciprocal teaching may be seen as a form of 'cognitive bootstrapping' to enable poor readers to escape the cycle of reading failure and engage more meaningfully in the process of reading. PMID- 12639277 TI - When can a lack of structure facilitate strategic processing of information? AB - BACKGROUND: Researchers examining the effectiveness of an elaboration strategy (answering 'why') for learning new information have been concerned with the familiarity of the materials and how that affects learning. It may be, however, that both how information is organised and familiarity impact on the potency of the strategy. AIMS: We examined the influence of presentation structures (i.e., organisation of information) on the effectiveness of an elaboration strategy. SAMPLES: All participants were undergraduates (78 females, 67 males) enrolled in a first-year psychology course. Fifteen students participated in Experiment One, 42 students in Experiment Two, and 88 students in Experiment Three. METHODS: In Experiment One, preference for conceptual organisation was assessed. In Experiments Two and Three, students answered 'why' questions when different presentation structures were used. RESULTS: Students' preferred method of organisation did not match the imposed structure found in past research, suggesting that students may have been restricted in their ability to process the information distinctively. Students who were presented with the information in a random order achieved the largest memory scores. CONCLUSIONS: When students have to reconstruct as well as encode the information, these added task demands provide an added benefit for learning. PMID- 12639278 TI - Multiple goals, motivation and academic learning. AB - BACKGROUND: The type of academic goals pursued by students is one of the most important variables in motivational research in educational contexts. Although motivational theory and research have emphasised the somewhat exclusive nature of two types of goal orientation (learning goals versus performance goals), some studies (Meece, 1994; Seifert, 1995, 1996) have shown that the two kinds of goals are relatively complementary and that it is possible for students to have multiple goals simultaneously, which guarantees some flexibility to adapt more efficaciously to various contexts and learning situations. AIM: The principal aim of this study is to determine the academic goals pursued by university students and to analyse the differences in several very significant variables related to motivation and academic learning. SAMPLE: Participants were 609 university students (74% women and 26% men) who filled in several questionnaires about the variables under study. METHOD: We used cluster analysis ('quick cluster analysis' method) to establish the different groups or clusters of individuals as a function of the three types of goals (learning goals, performance goals, and social reinforcement goals). By means of MANOVA, we determined whether the groups or clusters identified were significantly different in the variables that are relevant to motivation and academic learning. Lastly, we performed ANOVA on the variables that revealed significant effects in the previous analysis. RESULTS: Using cluster analysis, three groups of students with different motivational orientations were identified: a group with predominance of performance goals (Group PG: n = 230), a group with predominance of multiple goals (Group MG: n = 238), and a group with predominance of learning goals (Group LG: n = 141). CONCLUSIONS: Groups MG and LG attributed their success more to ability, they had higher perceived ability, they took task characteristics into account when planning which strategies to use in the learning process, they showed higher persistence, and used more deep learning strategies than did the students with predominance of performance goals (Group PG). On the other hand, Groups MG and PG took the evaluation criteria more into account when deciding which strategies to use in order to learn, and they attributed their failures more to luck than did Group LG. Students from Group MG attributed their success more to effort than did the other two groups and they attained higher achievement than Group PG. Group LG tended to attribute their failures more to lack of effort than did the other two groups. PMID- 12639279 TI - Construction and initial validation of the self-worth protection scale. AB - BACKGROUND: The self-worth theory of achievement motivation holds that in certain circumstances students stand to gain by deliberately withdrawing effort. When failure occurs despite effort, students are likely to conclude that failure resulted from lack of ability. Thus, withdrawing effort offers a defence against conclusions of low ability, thereby protecting self-worth. AIM: We undertook to assess the psychometric properties of the Self-Worth Protection Scale (SWPS). SAMPLE: Data were obtained from 243 participants (Study 1) and 411 participants (Study 2) enrolled in undergraduate psychology courses at a university in the United States. METHOD: We administered a number of scales, including the SWPS and scales assessing a fear of negative evaluation, academic self-esteem, uncertain global self-evaluations, self-handicapping, and causal uncertainty. RESULTS: Exploratory factor analysis indicated a three-factor solution (ability doubts, the importance of ability as a criterion of self-worth, and an avoidance orientation) utilising 33 of the original 44 items. A confirmatory factor analysis indicated that this three-factor solution was a poor fit of the data. After modifying the model, a confirmatory factor analysis indicated that a three factor solution with 26 of the original items and a higher order factor of self worth protection was an adequate fit of the data. Reliability measures were acceptable for the three subscales and total score. The total score of the SWPS was positively correlated with theoretically related constructs, demonstrating construct validity. CONCLUSIONS: The SWPS appears to be a psychometrically sound scale to assist in identifying individuals who manifest self-worth protection in achievement situations. PMID- 12639280 TI - Comprehension of arithmetic word problems by fifth-grade pupils: representations and selection of information. AB - BACKGROUND: Understanding word problems leads to the construction of different levels of representation. Some levels specify the elements which are indispensable for solving the problem (problem model, PM) and others specify the agents, actions and events in everyday concepts (situation model, SM). AIMS: By studying how the information is selected, we try to specify the nature of the representations constructed during the reading of a word problem: understanding a problem leads to the construction of two complementary levels of representation (PM and SM) or to the construction of only one representation (PM)? SAMPLE: Ninety-one fifth-grade pupils (mean age 10 years 9 months) took part in this study and were divided into two groups according to their mathematical ability. METHOD: As well as the information considered as indispensable for solving the problems (solving information), different types of information (situational information) were introduced into standard word problems. In a first task, participants were asked to select the information in order to 'make the word problem as short as possible' (locate the elements used for developing PM). In a second task, they were asked to select the information in order to 'make the word problem easier to understand' (determine whether the participants developed a SM). RESULTS: The participants successfully differentiated between the solving information and the situational information. An interaction was also observed between the type of information and the task. The mathematical ability of the participants was seen to have an influence on the selection of situational information. CONCLUSION: Understanding leads to the construction of two complementary representation levels: the problem model and the situation model. PMID- 12639281 TI - Making friends with your data: improving how statistics are conducted and reported. AB - AIM: This paper highlights some of the areas where there are problems with the way that statistics are conducted and reported in psychology journals. Recommendations are given for improving these problems. SAMPLE: The choice of topics is based largely on the questions that authors, reviewers, and editors have asked in recent years. The focus is on null hypothesis significance testing (NHST), choosing a statistical test, and what should be included in results sections. RESULTS: There are several ways to improve how statistics are reported. These should improve both the authors' and the readers' understanding of the data. CONCLUSIONS: Psychology as a discipline will improve if the way in which statistics are conducted and reported is improved. This will require effort from authors, scrutiny from reviewers, and stubbornness from editors. PMID- 12639282 TI - Effects of a Combination of Traditional Chinese Botanicals (Immune+) on the Secretion of Interleukin-1beta and Interferon-gamma by Peripheral Blood Mononuclear Cells. AB - The use of herbal and other botanical products, including those used extensively in traditional Chinese medicine, has increased dramatically in the last decade. Yet, little scientific research exists concerning their efficacy and safety. We examined the effects of Immune+, a combination of five botanicals frequently used in traditional Chinese medicine, on the production of two cytokines. For this purpose, unstimulated or phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells from healthy volunteers were incubated with different concentrations of Immune+. The secretion of interleukin-1beta (IL-1beta) and interferon-gamma (IFN-gamma) was measured after 72 hours of incubation. At the highest concentration tested (100 micro g/ml), Immune+ significantly increased the secretion of IL-1beta. Importantly, PHA alone had no effect on IL-1beta production, and the combination of PHA with Immune+ resulted in the same increase in IL-1beta production as seen with the botanical extract alone. Immune+ did not have any detectable effect on either unstimulated or PHA-stimulated IFN-gamma synthesis. These in vitro data support the concept that Immune+ may enhance human immune responses. PMID- 12639283 TI - Lycopene Content of Tomato Products: Its Stability, Bioavailability and In Vivo Antioxidant Properties. AB - Lycopene is a bioactive carotenoid present in many fruits and vegetables. Tomatoes constitute the major dietary source of lycopene. Recent evidence shows lycopene to be associated with several health benefits. However, very little information is available about the stability of lycopene and its bioavailability. Because tomatoes undergo extensive processing and storage before consumption, a study was conducted to evaluate the stability, isomeric form, bioavailability, and in vivo antioxidant properties of lycopene. Total lycopene and isomers were measured by spectrophotometry and high-performance liquid chromatography, respectively. Lycopene content of tomatoes remained unchanged during the multistep processing operations for the production of juice or paste and remained stable for up to 12 months of storage at ambient temperature. Moreover, subjecting tomato juice to cooking temperatures in the presence of corn oil resulted in the formation of the cis isomeric form, which was considered to be more bioavailable. Lycopene was absorbed readily from the dietary sources. Serum lipid and low-density lipoprotein oxidation were significantly reduced after the consumption of tomato products containing lycopene. PMID- 12639284 TI - MegaNatural((R)) Gold Grapeseed Extract: In Vitro Antioxidant and In Vivo Human Supplementation Studies. AB - Epidemiological studies have produced solid evidence that consumption of fruits results in a reduction of cancer and cardiovascular disease. Red wine has been hypothesized to reduce the risk of heart disease as a result of its antioxidant, endothelial relaxation, and antiplatelet aggregation mechanisms. We investigated a commercial California wine grapeseed extract, MegaNatural((R)) Gold, and compared it to grapes, grape juice, red wine, other grapeseed extracts, and commercial extracts with respect to antioxidant activity, using an oxidation reduction colorimetric assay. The quality of the antioxidants was determined with the use of an in vitro model of heart disease, a dose-response inhibition of low density lipoprotein oxidation. MegaNatural((R)) Gold had a superior quantity/quality antioxidant index compared with all other samples tested. In nine subjects given 600 mg of MegaNatural((R)) Gold, the plasma antioxidant activity as measured by the RANDOX assay was significantly increased by 12% after both 1 and 2 hours. This was equivalent to drinking 300 ml of red wine or consuming 1,250 mg of vitamin C, as found in a previous study. We then determined the dose-response human plasma bioavailability of one of the polyphenols in MegaNatural((R)) Gold (epicatechin) in nine human subjects after giving 200 and 300 mg of the product. The in vivo antioxidant activity was also measured. The higher dose was much more effective. A long-term human supplementation study with 2 x 300 mg/day of MegaNatural((R)) Gold was then done on 17 (9 normal and 8 hypercholesterolemic) subjects. Plasma cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol concentrations were significantly decreased in the subjects with high cholesterol. Triglycerides were significantly increased in the high-cholesterol subjects but were still in the normal range after supplementation. Plasma antioxidant capacity was significantly improved in the high-cholesterol subjects. There was no change in plasma lipids or antioxidant capacity in the normal subjects. PMID- 12639285 TI - Nicotinamide: A Nutritional Supplement that Provides Protection Against Neuronal and Vascular Injury. AB - In addition to functioning as an essential nutrient for cellular growth and maintenance, nicotinamide also may be an attractive therapeutic agent with efficacy demonstrated against free radical ischemic programmed cell death (PCD). Yet, the cellular mechanisms that mediate cellular PCD, as well as protection by nicotinamide, are considered to require further definition. In primary rat hippocampal neurons and rat cerebrovascular endothelial cells (ECs), cellular injury was determined through trypan blue dye exclusion, externalization of membrane phosphatidylserine (PS) residues, and activation of the mitogen activated protein kinase p38 through Western blot analysis. Nicotinamide was without cellular toxicity at concentrations lower than 50 mM in both neuronal and EC populations. Exposure to either anoxia or the nitric oxide (NO) donors sodium nitroprusside and NOC-9 significantly decreased neuronal and EC survival from approximately 85% to 38% and increased membrane PS exposure from approximately 10% to 80% over a 24-hour period. Pretreatment with nicotinamide (12.5 mM) prevented anoxic and NO cytodegeneration by significantly increasing survival and decreasing membrane PS expression. Protection by nicotinamide in both neurons and ECs appeared to be independent and downstream from p38 activation. Further investigations that define the cellular and molecular mechanisms employed by the nutrient nicotinamide may provide greater insight into the potential therapeutic targets that determine neuronal and vascular injury. PMID- 12639286 TI - Effects of Genistein Isoflavone (4',5',7-Trihydroxyisoflavone) and Dexamethasone on Functional Characteristics of Spermatozoa. AB - Caudal epididymal spermatozoa were used to study the influence of genistein isoflavone and dexamethasone (dxm) on the functional characteristics of spermatozoa. The effects of genistein alone and in combination with dxm on sperm motility, sperm morphology, spontaneous acrosome reaction (AcR), and ionophore A23187-induced AcR were investigated. The FITC-PSA/Hoechst 33258 staining procedure was used to assess sperm cell viability and AcR status and thus to differentiate between true AcR and acrosome degeneration. The overall results indicated that (1) lower doses of genistein alone, or in combination with dxm, did not significantly influence sperm motility or sperm morphology; (2) ionophore A23187 induced AcR in rat spermatozoa; (3) there appeared to be no direct correlation between sperm motility and AcR, (4) higher doses of genistein, alone or in combination with dxm, significantly interfered with percentage sperm motility and caused significant detachment of sperm heads but did not cause morphological defects; and (5) higher doses of genistein caused significant decrease in sperm acrosome reactivity with long duration of exposure. In view of the fact that sperm capacitation and AcR are physiological prerequisites for successful fertilization of oocytes, the findings suggest that chronic exposure of spermatozoa to high doses of genistein could be associated with infertility problems through suppression/inhibition of AcR and sperm motility. Dexamethasone did not appear to influence the effect of genistein on the functionality of postspermatogenic spermatozoa. PMID- 12639287 TI - Anticarcinogenic Activity of Strawberry, Blueberry, and Raspberry Extracts to Breast and Cervical Cancer Cells. AB - Freeze-dried fruits of two strawberry cultivars, Sweet Charlie and Carlsbad, and two blueberry cultivars, Tifblue and Premier were sequentially extracted with hexane, 50% hexane/ethyl acetate, ethyl acetate, ethanol, and 70% acetone/water at ambient temperature. Each extract was tested separately for in vitro anticancer activity on cervical and breast cancer cell lines. Ethanol extracts from all four fruits strongly inhibited CaSki and SiHa cervical cancer cell lines and MCF-7 and T47-D breast cancer cell lines. An unfractionated aqueous extract of raspberry and the ethanol extract of Premier blueberry significantly inhibited mutagenesis by both direct-acting and metabolically activated carcinogens. PMID- 12639288 TI - HIV-associated nephropathy: a review of the epidemiology and clinical course in the HAART era. PMID- 12639289 TI - Evaluation of pediatric HIV care provided in Ryan White CARE Act Title IV Women, Infants, Children, and Youth Clinics. AB - This evaluation examines the changing practices and outcomes of pediatric HIV care provided by the Ryan White Comprehensive AIDS Resources and Emergency (CARE) Act, Title IV grantees from 1996 through 1998--a period of rapidly changing medical practice within the United States. Using medical chart abstraction, 26 Title IV grantees reported information from the records of 525 HIV positive clients between the ages of 2 and 12. The chart abstractions covered medical care and case management provided to these clients including the number of clinical visits, use of antiretroviral therapy, use of laboratory tests such as CD4+ cell count (cells/mm3) and HIV-1 RNA viral load (copies/mL), enrollment in clinical trials, and receipt of opportunistic infection prophylaxis. Information on disease progression and hospitalization as well as client socio-demographic characteristics also is analyzed. Study results indicate that use of HIV combination therapy increased, while the occurrence of opportunistic infections and hospitalizations decreased. The increasing use of new pharmaceuticals during the study period indicates the feasibility of transferring information learned about HIV treatments from clinical trials to clinical practices that treat primarily Medicaid and pediatric populations. PMID- 12639290 TI - Sexual functioning among HIV-infected women. AB - The influence of HIV on sexual activity and functioning presents a unique set of issues. HIV transmissibility, partner disclosure, potential vertical transmission and, for some HIV-infected women, problems associated with illicit drug use, may affect sexuality. Few studies have examined HIV-infected women's sexuality and none have studied the relationship between HIV symptoms, HIV illness stage, quality of life, meaning of illness and sexual functioning. This descriptive study examines sexual activity, sexual functioning and safer sex behavior among 101 HIV-seropositive women. Results indicate that (1) the majority of women continue to be sexually active after testing HIV positive, (2) sexual functioning does not change as a result of HIV disease progression, and (3) few women report that HIV itself caused worsening of their sexual functioning. Those women with better mental health, more positive meaning attributed to life with HIV infection, better quality of life, fewer HIV-related symptoms and who never used injection drugs had higher levels of sexual functioning. Greater focus on sexual functioning as an integral part of clinical assessment is needed. Patterns of sexual behavior among HIV-infected women require continued study as we search for more effective means to prevent secondary HIV transmission. PMID- 12639291 TI - Evaluation of the care of women living with HIV/AIDS in Sao Paulo, Brazil. AB - To evaluate care provided to women living with HIV in Sao Paulo, Brazil, based on their own experiences, patients from HIV/AIDS reference clinics answered a questionnaire on circumstances of HIV testing, health personnel's attitudes at diagnosis, adherence to follow-up, services provided by care centers and access to laboratory monitoring. From September 1999 to February 2000, 1,068 women were interviewed. Most had over 8 years of education and reported having regular sexual partners, being mothers and family caregivers. They were diagnosed as HIV infected for 4.36 +/- 3.15 years. The majority had been referred to testing without counseling. Post-test counseling was provided depending on the testing center. Neither pre-test nor post-test counseling were associated to time since diagnosis. Some patients reported having felt indifference, discrimination or criticism at diagnosis, depending on where testing occurred. Compliance to medical follow-up was associated to adherence to antiretrovirals and to shorter time since diagnosis, but not with having a primary caregiver or with commuting time to the care center. Psychological support, nutrition care and oral health were women's less addressed needs. Access to gynecological care was associated with counseling on mother-to-child transmission and on safe sex. Access to CD4+ cell counts and HIV viral loads was concordant with guidelines, but the understanding of their meaning was incomplete. We conclude that women living with HIV/AIDS in Sao Paulo, Brazil have limited HIV risk perception and unmet care needs. Better training of professionals in charge of diagnosis and integrating women's health and family-planning actions to AIDS programs may enhance care. PMID- 12639292 TI - SOLO trial results released. PMID- 12639293 TI - IFN-alpha2b and thalidomide synergistically inhibit tumor-induced angiogenesis. AB - Angiogenesis is an absolute requirement for tumor growth and metastasis. The purpose of this study was to evaluate the antiangiogenic activity of interferon alpha2b (IFN-alpha2b) and thalidomide, as single agents and in combination. The murine dermis model was used to assess tumor-induced angiogenesis in nude mice. Human ACHN (renal), NIH-OVCAR-3 (ovarian), LNCaP (prostate), and SK-Mel-1 (melanoma) tumor cells were inoculated intradermally into the flanks of nude mice. IFN-alpha2b and thalidomide, administered daily, were effective inhibitors of angiogenesis induced by all four tumor types. The combination of IFN-alpha2b and thalidomide caused a synergistic decrease in mean vessel count in tumors that were resistant to the antiproliferative effects of IFN-alpha2b and thalidomide in vitro. This enhanced suppression of angiogenesis translated into synergistic antitumor activity in a xenograft model. Pegylated IFN-alpha (PEG-IFN-alpha2b) (10(6) U) administered once in 10 days was as effective as daily IFN-alpha2b treatment (10(6) U x 10 days). IFN-alpha2b and thalidomide have potentiated antiangiogenic activity when used in combination. A single dose of PEG-IFN alpha2b (10(6) U) was as effective at suppressing vessel growth as an equivalent dose of IFN-alpha2b given daily for 10 days. PMID- 12639294 TI - Pernicious anemia during IFN-alpha treatment for chronic hepatitis C. AB - Some latent diseases, such as immune disorders, can appear during interferon alpha (IFN-alpha) therapy. These disorders are difficult to predict because of their low prevalence in the general population. We describe a case of pernicious anemia (PA) in a patient affected by chronic hepatitis C and macrocytosis during IFN-alpha therapy. Hemoglobin (Hb) concentration reached 7.3 g/dl. Anti-intrinsic factor (IF) antibodies were present, but not antiparietal cell antibodies (APCA). Suspension of IFN-alpha and administration of vitamin B(12) resulted in normal Hb concentrations. This case is the first instance of early PA (at the second month of IFN therapy) in a patient affected by chronic hepatitis C. The only other case of PA in a patient affected by hepatitis C virus (HCV) infection occurred during the second year of maintenance IFN therapy. We recommend that particular attention be paid to such clinical and laboratory conditions as macrocytosis in administering IFN-alpha therapy for chronic hepatitis C. PMID- 12639295 TI - Deletion of IFN-gamma reduces fumonisin-induced hepatotoxicity in mice via alterations in inflammatory cytokines and apoptotic factors. AB - Fumonisin B(1) (FB(1)) produces species-specific and organ-specific toxicity, including equine leukoencephalomalacia, porcine pulmonary edema, and hepatic or renal damage in other animals. FB(1) causes inhibition of ceramide synthase, leading to accumulation of free sphingoid bases. We previously reported that such cytokines as tumor necrosis factor-alpha (TNF-alpha) modify FB(1)-induced hepatic apoptosis in male mice. FB(1) also caused induction of interferon-gamma (IFN gamma) in mouse liver, and, therefore, it was worthwhile to determine the role IFN-gamma plays in FB(1) toxicity. In the current study, male IFN-gamma-knockout (GKO) mice and their wild-type (WT) counterparts, C57BL/6J, were treated subcutaneously (s.c.) with 2.25 mg/kg/day of FB(1) for 5 days and sampled 1 day after the last injection. The levels of circulating liver enzymes were increased in WT animals but considerably less in GKO mice. Reduced hepatotoxicity in GKO mice was evident by histologic evaluation and enumeration of apoptotic cells. The induction of TNF-alpha and interleukin-12 (IL-12) p40 by FB(1) in liver was less in GKO mice compared with WT animals. The GKO mice also had a reduced accumulation of liver sphinganine than did WT mice after FB(1) treatment. Results suggested the implication of IFN-gamma in FB(1)-induced hepatotoxicity, which can be explained by a lack of TNF-alpha and IL-12 amplification in the liver of the GKO mice. In addition, the GKO mice had altered expression of various apoptotic and antiapoptotic factors in liver. These changes were accompanied by a greater number of proliferating cells in the liver of GKO mice after FB(1) treatment, which may also contribute to the reduced hepatotoxicity of FB(1) in GKO mice. Whereas the GKO mice show reduced sensitivity to FB(1) and FB(1) treatment elevates IFN-gamma expression, decreased hepatotoxicity to FB(1) could result from alterations in sphingolipid metabolism in the GKO strain. PMID- 12639296 TI - An IFN-beta-albumin fusion protein that displays improved pharmacokinetic and pharmacodynamic properties in nonhuman primates. AB - The long half-life and stability of human serum albumin (HSA) make it an attractive candidate for fusion to short-lived therapeutic proteins. Albuferon (Human Genome Sciences [HGS], Inc., Rockville, MD) beta is a novel recombinant protein derived from a gene fusion of interferon-beta (IFN-beta ) and HSA. In vitro, Albuferon beta displays antiviral and antiproliferative activities and triggers the IFN-stimulated response element (ISRE) signal transduction pathway. Array analysis of 5694 independent genes in Daudi-treated cells revealed that Albuferon beta and IFN-beta induce the expression of an identical set of 30 genes, including 9 previously not identified. In rhesus monkeys administered a dose of 50 microg/kg intravenously (i.v.) or subcutaneously (s.c.) or 300 microg/kg s.c., Albuferon beta demonstrated favorable pharmacokinetic properties. Subcutaneous bioavailability was 87%, plasma clearance at 4.7-5.7 ml/h/kg was approximately 140-fold lower than that of IFN-beta, and the terminal half-life was 36-40 h compared with 8 h for IFN-beta. Importantly, Albuferon beta induced sustained increases in serum neopterin levels and 2',5' mRNA expression. At a molar dose equivalent to one-half the dose of IFN-beta, Albuferon beta elicited comparable neopterin responses and significantly higher 2',5'-OAS mRNA levels in rhesus monkeys. The enhanced in vivo pharmacologic properties of IFN-beta when fused to serum albumin suggest a clinical opportunity for improved IFN-beta therapy. PMID- 12639297 TI - Comparative DNA sequence analysis of mouse and human CC chemokine gene clusters. AB - The CC chemokines are a closely related subfamily of the chemokine superfamily. Most of the CC chemokine genes form a cluster on chromosome 11 in mice and chromosome 17 in humans. To date, 11 and 16 functional genes have been localized within the mouse and human clusters, respectively. Notably, some of the genes within these clusters appear to have no counterparts between the two species, and the orthologous relationships of some of the genes are difficult to establish solely on the basis of amino acid similarity. In this study, we have taken a comparative genomic approach to reveal some of the features that may be involved in the dynamic evolution of these gene clusters. We sequenced a 122-kb region containing five chemokine genes of the mouse CC cluster. This mouse sequence was combined with those determined by the Mouse Genome Sequencing Project, and the entire sequence of the mouse CC cluster was compared with that of the corresponding cluster in the human genome by percent identity plot and dot-plot analyses. Although no additional chemokine genes have been found in these clusters, our analysis has revealed that numerous gene rearrangements have occurred even after the diversification of rodents and primates, resulting in several species-specific chemokine genes and pseudogenes. In addition, phylogenetic analysis and comparison of the genomic sequences unambiguously identified the orthologous relationships of some of the chemokine genes in the mouse and human CC gene clusters. PMID- 12639298 TI - Antibody and interleukin-12 treatment in murine models of encephalitogenic flavivirus (St. Louis encephalitis, tick-borne encephalitis) and alphavirus (Venezuelan equine encephalitis) infection. AB - Early and sustained treatment with interleukin-12 (IL-12) ameliorated disease in a mouse model of infection with the encephalitogenic flavivirus, St. Louis encephalitis virus (SLEV, Japanese encephalitis serogroup). However, this effect was not reproduced in murine infections with either the flavivirus tick-bore encephalitis virus (TBEV) or the alphavirus Venezuelan equine encephalitis virus (VEEV). IL-12 exacerbated TBEV disease when used in conjunction with monoclonal antibody (mAb), suggesting an enhancement of immunopathology, and was without clinical effects in VEEV infection. These data confirm the need to fully understand the pathogenesis of viral infection before cytokine intervention may be employed as a broad-spectrum antiviral therapy. PMID- 12639299 TI - Minimal change disease in a patient receiving IFN-alpha therapy for chronic hepatitis C virus infection. AB - Chronic hepatitis C virus (HCV) infection is associated with several extrahepatic syndromes. The principal types of renal disorders associated with chronic HCV infection are cryoglobulinemia or noncryoglobulinemic membranoproliferative glomerulonephritis (MPGN). Interferon-alpha (IFN-alpha) may precipitate or exacerbate the occurrence of MPGN. Our patient was a 32-year-old man who tested positive for HCV in July 1997. The patient was treated with IFN-alpha in another medical center for 6 months because his liver biopsy showed chronic active hepatitis. In December 1998, he applied to our clinic for a follow-up examination. The level of aspartate aminotransferase (AST) was 44 U/L, and that of alanine aminotransferase (ALT) was 69 U/L. HCV RNA was positive in serum, and chronic HCV infection was detected by liver biopsy. IFN-alpha therapy (5 million U/day) was administered for 6 months longer. In May 1999, the patient came to our polyclinic with edema of the feet and legs. We detected proteinuria, serum cholesterol of 269 mg/dl, AST of 50 U/L, ALT of 41 U/L, serum total protein of 3.4 g/dl, serum albumin of 1.2 g/dl, positive cryoglobulin, and urine protein of 9.84 g/day. Cryoglobulinemic MPGN was suspected and kidney biopsy was performed, resulting in a diagnosis of minimal change disease (MCD). PMID- 12639300 TI - Stability of mRNA/cationic lipid lipoplexes in human and rat cerebrospinal fluid: methods and evidence for nonviral mRNA gene delivery to the central nervous system. AB - Clinical applications of gene therapy require advances in gene delivery systems. Although numerous clinical trials are already underway, the ultimate success of gene therapies will depend on gene transfer vectors that facilitate the expression of a specific gene at therapeutic levels in the desired cell populations without eliciting cytotoxicity. In clinical applications for which transient expression is desirable, mRNA delivery is of particular interest. We have shown cationic lipid-mediated mRNA delivery to be feasible, efficient, and reproducible in vitro. mRNA delivery to the cerebrospinal fluid (CSF) in vivo would provide a means of vector distribution throughout the central nervous system (CNS). This study examined the functional integrity and protection from degradation of mRNA/cationic complexes (lipoplexes) in human cerebrospinal fluid (hCSF) in vitro and expression of these lipoplexes in vivo. Results obtained from gel electrophoresis indicate that cationic lipids protect mRNA transcripts from RNases in hCSF for at least 4 hr. This is in contrast to the total disappearance of nonlipid-complexed mRNA in less than 5 min. We confirmed the importance of RNase activity by incubating mRNA transcripts encoding luciferase or green fluorescent protein (GFP) in hCSF to which RNase inhibitors had been added. After incubation, these solutions were used to transfect Chinese hamster ovary (CHO) cells in vitro. Next, assays for both GFP and luciferase were used to demonstrate functional integrity and translation of the mRNA transcripts. Finally, we delivered in vitro transcribed mRNA vectors encoding for Hsp70 and luciferase to the lateral ventricle of the rat in a series of preliminary in vivo experiments. Initial immunohistochemistry analysis demonstrates that the distribution, uptake, and expression of reporter sequences using lipid-mediated mRNA vector delivery is extensive, as we earlier reported using similar methods with DNA vectors but that the expression may be less intense. Expression was noted in coronal sections throughout the rat brain, confirming the potential for lipid-mediated mRNA delivery to the CNS. These findings confirm that complexing mRNA with cationic lipid before exposure to CSF confers protection against RNase activity, facilitating distribution, cellular uptake, and expression of mRNA delivered into the CNS. PMID- 12639301 TI - DNA sequences in cationic lipid:pDNA-mediated systemic toxicities. AB - Systemic delivery of synthetic gene transfer vectors such as cationic lipid:plasmid DNA (pDNA) complexes elicits a range of acute physiologic responses, which in the context of therapeutic gene delivery represent dose limiting toxicities. The most prominent responses are transient leukopenia, thrombocytopenia, serum transaminase elevations, and elevations of proinflammatory cytokines such as interferon-gamma (IFN-gamma), interleukin-12 (IL-12), and tumor necrosis factor-alpha (TNF-alpha). The unmethylated CpG sequences present in plasmid DNA have been implicated as a major cause of the robust cytokine response that follows systemic administration of cationic lipid:pDNA complexes. However, the factors causing the additional significant toxicities (leukopenia, thrombocytopenia, and serum transaminase elevations) recently shown to be associated with vector administration have not been defined. We show here that DNA sequences, such as immune stimulatory CpG sequences, play a significant role in inducing the additional acute toxicities associated with cationic lipid:pDNA complex administration. Importantly, while methylating these CpG sequences results in greatly reduced cytokine levels, this modification does not eliminate their ability to generate the other systemic toxicities. Examples of non-CpG DNA sequences that induce distinct toxicity profiles when administered systemically in the form of cationic lipid:DNA complexes are also identified. Taken together, these results imply that specific DNA sequences are responsible for a significant portion of the systemic toxicities observed after administration of cationic lipid:pDNA complexes. PMID- 12639302 TI - Optimization of cis-acting elements for gene expression from nonviral vectors in vivo. AB - While naked DNA gene transfer in vivo usually results in transient gene expression, in some cases long-term transgene expression can be achieved. Here we demonstrate that cis-acting DNA elements flanking the transgene expression cassette and components in the plasmid backbone can significantly influence expression levels from nonviral vectors. To demonstrate this, we administered our most robust human coagulation factor IX (hFIX) expression cassette placed in two different plasmid backbones, into the livers of mice, by hydrodynamic transfection. We found that placing the expression cassette within a minimal plasmid vector pHM5, a modified version of pUC19, resulted in 10 times higher serum hFIX expression levels (up to 20000 ng/ml, 400% of normal hFIX serum levels), compared to a pBluescript backbone. To optimally increase expression levels from a nonviral vector, we added matrix attachment regions (MARs) as cis acting DNA elements flanking the hFIX expression cassette. We detected five fold higher hFIX expression levels in vivo for up to 1-year posttransfection from a vector that contained the chicken MAR from the lysozyme locus. Together, the present work demonstrates that in addition to the transgene expression cassette, cis-acting DNA elements within and outside of the plasmid backbone need to be evaluated to achieve optimal expression levels in a nonviral gene therapy approach. PMID- 12639304 TI - Optimization of the generation and propagation of gutless adenoviral vectors. AB - Adenoviral vectors devoid of all viral coding regions are referred to by many names, including gutless vectors. Gutless vectors display reduced toxicity and immunogenicity, increased duration of transgene expression, and increased coding capacity compared to early generation vectors, which contain the majority of the viral backbone genes. However, the production of gutless vectors at a scale and purity suitable for clinical use has limited the utility of this technology. In this work we describe the optimization of the production of gutless vectors. We constructed an improved helper virus and generated an alternative gutless vector producer cell line, PERC6-Cre. We demonstrated increased gutless vector yields, minimal helper virus contamination, and no replication-competent adenovirus contamination using the optimized system. Furthermore, the PERC6-Cre cells were adapted to serum-free suspension culture and high-titer gutless vector preparations were produced using bioreactor technology, suggesting the feasibility of gutless vector scale-up for clinical use. Finally, we observed that helper virus lacking a packaging signal could be packaged at a low frequency, revealing an inherent limitation to the differential packaging strategy for gutless vector propagation. PMID- 12639303 TI - Phase I dose escalation clinical trial of adenovirus vector carrying osteocalcin promoter-driven herpes simplex virus thymidine kinase in localized and metastatic hormone-refractory prostate cancer. AB - Osteocalcin (OC), a major noncollagenous bone matrix protein, is expressed prevalently in prostate cancer epithelial cells, adjacent fibromuscular stromal cells, and osteoblasts in locally recurrent prostate cancer and prostate cancer bone metastasis [Matsubara, S., Wada, Y., Gardner, T.A., Egawa, M., Park, M.S., Hsieh, C.L., Zhau, H.E., Kao, C., Kamidono, S., Gillenwater, J.Y., and Chung, L.W. (2001). Cancer Res. 61, 6012-6019]. We constructed an adenovirus vector carrying osteocalcin promoter-driven herpes simplex virus thymidine kinase (Ad-OC hsv-TK) to cotarget prostate cancer cells and their surrounding stromal cells. A phase I dose escalation clinical trial of the intralesional administration of Ad OC-hsv-TK followed by oral valacyclovir was conducted at the University of Virginia (Charlottesville, VA) in 11 men with localized recurrent and metastatic hormone-refractory prostate cancer (2 local recurrent, 5 osseous metastasis, and 4 lymph node metastasis) in order to determine the usefulness of this vector for the palliation of androgen-independent prostate cancer metastasis. This is the first clinical trial in which therapeutic adenoviruses are injected directly into prostate cancer lymph node and bone metastasis. Results show that (1). all patients tolerated this therapy with no serious adverse events; (2). local cell death was observed in treated lesions in seven patients (63.6%) as assessed by TUNEL assay, and histomorphological change (mediation of fibrosis) was detected in all posttreated specimens; (3). one patient showed stabilization of the treated lesion for 317 days with no alternative therapy. Of the two patients who complained of tumor-associated symptoms before the treatment, one patient with bone pain had resolution of pain, although significant remission of treated lesions was not observed by image examination; (4). CD8-positive T cells were predominant compared with CD4-positive T cells, B cells (L26 positive), and natural killer cells (CD56 positive) in posttreated tissue specimens; (5). levels of HSV TK gene transduction correlated well with coxsackie-adenovirus receptor expression but less well with the titers of adenovirus injected; and (6). intrinsic OC expression and the efficiency of HSV TK gene transduction affected the levels of HSV TK protein expression in clinical specimens. Our data suggest that this form of gene therapy requires further development for the treatment of androgen-independent prostate cancer metastasis although histopathological and immunohistochemical evidence of apoptosis was observed in the specimens treated. Further studies including the development of viral delivery will enhance the efficacy of Ad-OC-hsv-TK. PMID- 12639305 TI - In vivo quantitative noninvasive imaging of gene transfer by single-photon emission computerized tomography. AB - Systems aimed at detecting gene expression noninvasively in vivo are desirable for evaluating the outcome of gene transfer in clinical trials. Several approaches have been exploited using magnetic resonance imaging and spectroscopy ((31)P MRS), positron emission tomography (PET), single-photon emission tomography (SPECT), and detection of bioluminescent signals. An ideal system is based on transfer of a marker gene, the activity of which can be detected against a background from the target tissue without interfering with normal physiology or eliciting an immune response. The majority of approaches described to date use genes encoding a nonmammalian protein that can elicit immune responses or a transmembrane receptor as a marker gene whose ectopic expression may cause aberrant signaling in the target cell through binding to endogenous ligands. The dopamine transporter (DAT) is normally expressed at high levels, mainly in the dopaminergic neurons of the central nervous system. We previously synthesized a radioactive ligand, [(99m)Tc]TRODAT-1, that binds with high affinity to the dopamine transporter, allowing for SPECT imaging of the striatum in normal control subjects and individuals affected with Parkinson's disease. Here we describe a strategy to monitor gene transfer based on adeno-associated viral vector (AAV)-mediated transduction of DAT in murine muscle followed by [(99m)Tc]TRODAT-1 imaging by SPECT of cells expressing the transgene. We show that quantitative, noninvasive imaging of gene transfer is successfully achieved in vivo, using a single-photon computed tomography camera. This system employs a reporter gene encoding a mammalian protein that is absent in most tissues, has no enzymatic activity, and does not activate intracellular pathways. This should be useful to monitor gene transfer in the settings of gene therapy. PMID- 12639306 TI - Efficiency of onco-retroviral and lentiviral gene transfer into primary mouse and human B-lymphocytes is pseudotype dependent. AB - B lymphocytes are attractive targets for gene therapy of genetic diseases associated with B-cell dysfunction and for immunotherapy. Transduction of B lymphocytes was evaluated using green fluorescent protein (GFP)-encoding onco retroviral and HIV-derived lentiviral vectors which were pseudotyped with ecotropic, amphotropic or vesicular stomatitis virus (VSV-G) envelopes. Transduction of mouse B lymphocytes activated with lipopolysaccharides (LPS) or by cross-linking CD40 in conjunction with interleukin-4 (IL-4) was significantly more efficient (p < 0.003) with ecotropic (11%) than with VSV-G pseudotyped onco retroviral vectors (1%). Using high-titer cell-free ecotropic viral supernatant or by coculture with ecotropic onco-retroviral vector-producing cells, transduction efficiency increased significantly (p < 0.001) to approximately 50%, whereas transduction efficiency by coculture with VSV-G pseudotyped vector producing cells remained low (< 2%). Similarly, transduction of mouse B lymphocytes was significantly more efficient (twofold, p < 0.01) with the ecotropic (7%) than with the VSV-G pseudotyped lentiviral vectors although gene transfer efficiency remained low because of dose-limiting toxicity of the concentrated vector preparations on the LPS-activated murine B cells. Consistent with murine B-cell transduction, human B cells activated with CD40L and IL-4 were also found to be relatively refractory to VSV-G pseudotyped onco-retroviral vectors (< 1%). However, higher transduction efficiencies could be achieved in activated primary human B lymphocytes using VSV-G pseudotyped lentiviral vectors instead (5%-6%). Contrary to the significant increase in mouse B-cell transduction efficiency with ecotropic vectors, the use of amphotropic onco retroviral or lentiviral vectors did not increase transduction efficiency in primary human B cells. The present study shows that the transduction efficiency of onco-retroviral and lentiviral vectors in human and mouse B lymphocytes is pseudotype-dependent and challenges the widely held assumption that VSV-G pseudotyping facilitates gene transfer into all cell types. PMID- 12639307 TI - Improved herpes simplex virus type 1 amplicon vectors for proportional coexpression of positron emission tomography marker and therapeutic genes. AB - For the development of efficient and safe gene therapy protocols for clinical application it is desirable to determine the tissue dose of vector-mediated therapeutic gene expression noninvasively in vivo. The herpes simplex virus type 1 thymidine kinase gene (HSV-1-tk) has been shown to function as a marker gene for the direct noninvasive in vivo localization of thymidine kinase (TK) expression by positron emission tomography (PET). Using bicistronic or multicistronic gene-expressing cassettes with tk as the PET marker gene, the quantitative analysis of tk gene expression may indirectly indicate the distribution and the level of expression of linked and proportionally coexpressed genes. Here, we describe the construction and functional evaluation of HSV-1 amplicon vectors mediating proportional coexpression of HSV-1-tk as PET marker gene and the enhanced green fluorescent protein gene (gfp) as proof of principle and cell culture marker gene and the Escherichia coli cytosine deaminase (cd) as therapeutic gene. Several double-/triple-gene constructs expressing HSV-1-tk, gfp, and E. coli cd were engineered based on gene fusion or the use of an internal ribosome entry site (IRES). Functional analysis in cell culture (green fluorescent protein [GFP] fluorescence and sensitivity to the prodrugs ganciclovir [GCV] and 5-fluorocytosine [5-FC]) and Western blots were carried out after infection of proliferating rat 9L gliosarcoma and human Gli36 glioma cells with helper virus-free packaged HSV-1 amplicon vectors. To study the ability of PET to differentiate various levels of tk expression noninvasively in vivo, retrovirally transduced and selected populations of rat F98 and human Gli36dEGFR glioma cells with defined levels of proportionally coexpressed tk and gfp genes were grown as subcutaneous tumors in nude rats and nude mice, and tk imaging by PET was performed. To study HSV-1 amplicon vector-mediated gene coexpression in vivo, HSV-1 amplicon vectors bearing coexpression constructs were injected (4 x 10(7) to 1 x 10(8) transducing units) into subcutaneously growing Gli36dEGFR gliomas in nude animals, and tk imaging was performed 24 hr later. All vector constructs mediated GFP expression and sensitized 9L and Gli36 cells toward GCV- and 5-FC-mediated cell killing in a drug dose-dependent manner, respectively. The levels of gene expression varied depending on the location of the genes within the constructs indicating the influence of the IRES on the level of expression of the second gene. Moreover, functional proportional coexpression of the PET marker gene HSV-1-tk and the linked therapeutic E. coli cd gene was observed. In selected tumor cell populations, subtle IRES-dependent differences of tk gene expression could be noninvasively distinguished by PET with good correlation between quantitative assays for IRES-dependent attenuated GFP and TK expression in culture and in vivo. After infection of subcutaneously growing gliomas with HSV-1 amplicon vectors, various levels of TK expression were found ranging from 0.011-0.062 percentage injected dose per gram (%ID/g). These values were 4.0- to 5.7-fold lower than positive control tumor cells. TK expression could be imaged by PET in vivo even with the tk gene located at the weak position downstream from the IRES. In conclusion, these HSV-1 amplicon vectors carrying HSV-1-tk as PET marker gene and any linked therapeutic gene will serve an indirect noninvasive assessment of the distribution of therapeutic gene expression by PET. Monitoring the correlation between primary transduction and therapeutic efficiency of a given vector is highly desirable for the development of safe and efficient gene therapy and vector application protocols in clinical applications. PMID- 12639311 TI - A novel approach to the use of genetically modified herbicide tolerant crops for environmental benefit. AB - The proposed introduction of genetically modified herbicide tolerant (GMHT) crops, with claims of improved weed control, has prompted fears about possible environmental impacts of their widespread adoption, particularly on arable weeds, insects and associated farmland birds. In response to this, we have developed a novel weed-management system for GMHT sugar beet, based on band spraying, which exploits the flexibility offered by the broad-spectrum partner herbicides. Here, we show the results from two series of field experiments which, taken together, demonstrate that, by using this system, crops can be managed for enhanced weed and insect biomass without compromising yield, thus potentially offering food and shelter to farmland birds and other wildlife. These results could be applicable widely to other row crops, and indicate that creative use of GMHT technology could be a powerful tool for developing more sustainable farming systems in the future. PMID- 12639312 TI - Inverting the hourglass: quantitative evidence against the phylotypic stage in vertebrate development. AB - The concept of a phylotypic stage, when all vertebrate embryos show low phenotypic diversity, is an important cornerstone underlying modern developmental biology. Many theories involving patterns of development, developmental modules, mechanisms of development including developmental integration, and the action of natural selection on embryological stages have been proposed with reference to the phylotypic stage. However, the phylotypic stage has never been precisely defined, or conclusively supported or disproved by comparative quantitative data. We tested the predictions of the 'developmental hourglass' definition of the phylotypic stage quantitatively by looking at the pattern of developmental-timing variation across vertebrates as a whole and within mammals. For both datasets, the results using two different metrics were counter to the predictions of the definition: phenotypic variation between species was highest in the middle of the developmental sequence. This surprising degree of developmental character independence argues against the existence of a phylotypic stage in vertebrates. Instead, we hypothesize that numerous tightly delimited developmental modules exist during the mid-embryonic period. Further, the high level of timing changes (heterochrony) between these modules may be an important evolutionary mechanism giving rise to the diversity of vertebrates. The onus is now clearly on proponents of the phylotypic stage to present both a clear definition of it and quantitative data supporting its existence. PMID- 12639313 TI - A comparative study of white blood cell counts and disease risk in carnivores. AB - In primates, baseline levels of white blood cell (WBC) counts are related to mating promiscuity. It was hypothesized that differences in the primate immune system reflect pathogen risks from sexually transmitted diseases (STDs). Here, we test for the generality of this result by examining hypotheses involving behavioural, ecological and life-history factors in carnivores. Again, we find a significant correlation in carnivores between mating promiscuity and elevated levels of WBC counts. In addition, we find relationships with measures of sociality, substrate use and life-history parameters. These comparative results across independent taxonomic orders indicate that the evolution of the immune system, as represented by phylogenetic differences in basal levels of blood cell counts, is closely linked to disease risk involved with promiscuous mating and associated variables. We found only limited support for an association between the percentage of meat in the diet and WBC counts, which is consistent with the behavioural and physiological mechanisms that carnivores use to avoid parasite transmission from their prey. We discuss additional comparative questions related to taxonomic differences in disease risk, modes of parasite transmission and implications for conservation biology. PMID- 12639314 TI - Variation in immune defence as a question of evolutionary ecology. AB - The evolutionary-ecology approach to studying immune defences has generated a number of hypotheses that help to explain the observed variance in responses. Here, selected topics are reviewed in an attempt to identify the common problems, connections and generalities of the approach. In particular, the cost of immune defence, response specificity, sexual selection, neighbourhood effects and questions of optimal defence portfolios are discussed. While these questions still warrant further investigation, future challenges are the development of synthetic concepts for vertebrate and invertebrate systems and also of the theory that predicts immune responses based on a priori principles of evolutionary ecology. PMID- 12639315 TI - Variable responses to large-scale climate change in European Parus populations. AB - Spring temperatures in temperate regions have increased over the past 20 years and many organisms have responded to this increase by advancing the timing of their growth and reproduction. However, not all populations show an advancement of phenology. Understanding why some populations advance and others do not will give us insight into the possible constraints and selection pressures on the advancement of phenology. By combining two decades of data on 24 populations of tits (Parus sp.) from six European countries, we show that the phenological response to large-scale changes in spring temperature varies across a species' range, even between populations situated close to each other. We show that this variation cannot be fully explained by variation in the temperature change during the pre- and post-laying periods, as recently suggested. Instead, we find evidence for a link between rising temperatures and the frequency of second broods, which results in complex shifts in the laying dates of first clutches. Our results emphasize the need to consider links between different life-history parameters in order to predict the ecological consequences of large-scale climate changes. PMID- 12639316 TI - Magnetic cues and time of season affect fuel deposition in migratory thrush nightingales (Luscinia luscinia). AB - Bird migration requires high energy expenditure, and long-distance migrants accumulate fat for use as fuel during stopovers throughout their journey. Recent studies have shown that long-distance migratory birds, besides accumulating fat for use as fuel, also show adaptive phenotypic flexibility in several organs during migration. The migratory routes of many songbirds include stretches of sea and desert where fuelling is not possible. Large fuel loads increase flight costs and predation risk, therefore extensive fuelling should occur only immediately prior to crossing inhospitable zones. However, despite their crucial importance for the survival of migratory birds, both strategic refuelling decisions and variation in phenotypic flexibility during migration are not well understood. First-year thrush nightingales (Luscinia luscinia) caught in the early phase of the onset of autumn migration in southeast Sweden and exposed to a magnetic treatment simulating a migratory flight to northern Egypt increased more in fuel load than control birds. By contrast, birds trapped during the late phase of the onset of autumn migration accumulated a high fuel load irrespective of magnetic treatment. Furthermore, early birds increased less in flight-muscle size than birds trapped later in autumn. We suggest that the relative importance of endogenous and environmental factors in individual birds is affected by the time of season and by geographical area. When approaching a barrier, environmental cues may act irrespective of the endogenous time programme. PMID- 12639318 TI - Cuticular lipids as trail pheromone in a social wasp. AB - We investigated the origin and composition of the chemical trail of the common yellow jacket Vespula vulgaris L. (Vespidae) and found that an artificial trail made from an extract of cuticular lipids from V. vulgaris foragers was biologically as active as a trail laid naturally by the foragers. Chemical analysis of natural trail extracts and the behaviourally active cuticular extracts by coupled gas chromatography-mass spectrometry revealed that the majority of cuticular hydrocarbons were also present in the trail extract at similar ratios. Thus, bioassay data and chemical analysis provide strong evidence that these cuticular hydrocarbons act as a trail pheromone in V. vulgaris. PMID- 12639317 TI - Subordinate superb fairy-wrens (Malurus cyaneus) parasitize the reproductive success of attractive dominant males. AB - Explanations of cooperative breeding have largely focused on the indirect benefits philopatric offspring gain from investing in kin. However, recent molecular studies have revealed that in many species subordinates provision unrelated offspring. This has led to the re-evaluation of the direct and indirect benefits of helping behaviour. In this study, we used microsatellite genotyping to assess the extra-group reproductive success of subordinate superb fairy-wrens (Malurus cyaneus), a species with extremely high rates of extra-group paternity. Extra-group subordinate males sired 10.2% (193 out of 1895) of all offspring sampled between 1993 and 2000 and 21.4% (193 out of 901) of all illegitimate offspring sired by known males. The extra-group success of subordinates was greatly influenced by the attractiveness of their dominant male. Subordinates of attractive dominants sired more extra-group young than did average dominants. Evidence suggests that mate choice in superb fairy-wrens is error-prone and subordinates can gain direct reproductive benefits through parasitizing the reproductive success of attractive dominants. PMID- 12639319 TI - Chasing a dummy target: smooth pursuit and velocity control in male blowflies. AB - Male blowflies chase and catch other flies in fast acrobatic flights. To unravel the underlying control system, we presented a black moving sphere instead of a real fly as a pursuit target. By varying the size and speed of the target, we were able to systematically analyse the decisive visual determinants that guide chasing behaviour. Flies pursue targets of a wide range of sizes and velocities. The percentage of pursuits resulting in target capture decreases with increasing target size and speed. Chasing male flies adjust their forward velocity depending on the retinal size of the target, indicating that retinal size is a relevant input variable of the control system. The chasing fly focuses the target with great accuracy in the frontal part of its visual field by means of a smooth pursuit control system using the retinal position of the target to determine the flight direction. We conclude that for a comprehensive understanding of chasing control different time lags in the control systems of angular and forward velocity together with the impact of inertia on fly movements need to be taken into account. PMID- 12639320 TI - Geographical range and speciation in fossil and living molluscs. AB - The notion of a positive relation between geographical range and speciation rate or speciation probability may go back to Darwin, but a negative relation between these parameters is equally plausible. Here, we test these alternatives in fossil and living molluscan taxa. Late Cretaceous gastropod genera exhibit a strong negative relation between the geographical ranges of constituent species and speciation rate per species per million years; this result is robust to sampling biases against small-bodied taxa and is not attributable to phylogenetic effects. They also exhibit weak inverse or non-significant relations between geographical range and (i) the total number of species produced over the 18 million year timeframe, and (ii) the number of species in a single timeplane. Sister-group comparisons using extant molluscan species also show a non-significant relation between median geographical range and species richness of genera. These results support the view that the factors promoting broad geographical ranges also tend to damp speciation rates. They also demonstrate that a strong inverse relation between per-species speciation rate and geographical range need not be reflected in analyses conducted within a single timeplane, underscoring the inadequacy of treating net speciation as a proxy for raw per-taxon rates. PMID- 12639321 TI - Ribbon worm relationships: a phylogeny of the phylum Nemertea. AB - We present the most extensive phylogenetic analysis to date, to our knowledge, of higher-level nemertean relationships, based on sequence data from four different genes (the nuclear genes for nuclear large subunit rRNA (28S rRNA) and histone H3 (H3), and the mitochondrial genes for mitochondrial large subunit rRNA (16S rRNA) and cytochrome c oxidase subunit I (COI)). Well-supported clades are, in general, compatible with earlier, more limited, analyses, and current classification is largely in agreement with our results, although there are some notable exceptions. Bdellonemertea (represented by Malacobdella) is found to be a part of Monostilifera, and Polystilifera is the monophyletic sister group to Monostilifera. Cratenemertidae is the sister group to the remaining monostiliferans (including Malacobdella), a group to which we apply the new name Distromatonemertea. Heteronemertea is monophyletic and forms a clade with Hubrechtella; for this clade we introduce the name Pilidiophora. Finally, Pilidiophora and Hoplonemertea (with Malacobdella) form a monophyletic group, and we introduce the name Neonemertea to refer to this group. Palaeonemertea is found to be non-monophyletic and basal among nemerteans. PMID- 12639322 TI - Recent habitat fragmentation caused by major roads leads to reduction of gene flow and loss of genetic variability in ground beetles. AB - Although habitat fragmentation is suspected to jeopardize the long-term survival of many species, few data are available on its impact on the genetic variability of invertebrates. We assess the genetic population structure of the flightless ground beetle Carabus violaceus L., 1758 in a Swiss forest, which is divided into several fragments by a highway and two main roads. Eight samples were collected from different forest fragments and analysed at six microsatellite loci. The largest genetic differentiation was observed between samples separated by roads and in particular by the highway. The number of roads between sites explained 44% of the variance in pairwise F(ST) estimates, whereas the age of the road and the geographical distance between locations were not significant factors. Furthermore, a comparison of allelic richness showed that the genetic variability in a small forest fragment isolated by the highway was significantly lower than in the rest of the study area. These findings strongly support the hypothesis that large roads are absolute barriers to gene flow in C. violaceus, which may lead to a loss of genetic variability in fragmented populations. PMID- 12639323 TI - Genetic variation over space and time: analyses of extinct and remnant lake trout populations in the Upper Great Lakes. AB - Lake trout (Salvelinus namaycush) in the upper Laurentian Great Lakes of North America experienced striking reductions in abundance and distribution during the mid-twentieth century. Complete collapse of populations was documented for Lake Michigan, and a few remnant populations remained only in lakes Huron and Superior. Using DNA obtained from historical scale collections, we analysed patterns of genetic diversity at five microsatellite loci from archived historical samples representing 15 populations (range 1940-1959) and from three contemporary remnant populations across lakes Huron and Superior (total n = 893). Demographic declines in abundance and the extirpation of native lake trout populations during the past 40 years have resulted in the loss of genetic diversity between lakes owing to extirpation of Lake Michigan populations and a temporal trend for reduction in allelic richness in the populations of lakes Superior and Huron. Naturally reproducing populations in Lake Superior, which had been considered to be remnants of historical populations, and which were believed to be responsible for the resurgence of lake trout numbers and distribution, have probably been affected by hatchery supplementation. PMID- 12639324 TI - Spatio-temporal dynamics of the grey-sided vole in Hokkaido: identifying coupling using state-based Markov-chain modelling. AB - Explaining synchronization of cyclical or fluctuating populations over geographical regions presents ecologists with novel analytical challenges. We have developed a method to measure synchrony within spatial-temporal datasets of population densities applicable to both periodic and irregularly fluctuating populations. The dynamics of each constituent population is represented by a discrete Markov model. The state of a population trajectory at each time-point is classified as one of 'increase', 'decrease', 'peak' or 'trough'. The set of populations at any time-point is characterized by the frequency distribution of these different states, and the time-evolution of this frequency distribution used to test the hypothesis that the dynamics of each population proceeds independently of the others. The analysis identifies years in which population coupling results in synchronous states and onto which states the system converges, and identifies those years in which synchrony remains high but is accounted for by coupling observed in previous years. It also enables identification of which pairs of sites show the highest levels of coupling. Applying these methods to populations of the grey-sided vole on Hokkaido reveals them to be fluctuating in greater synchrony than would be expected from independent dynamics, and that this level of synchrony is maintained through intermittent coupling acting in ca. 1 year in four or five. High synchrony occurs between sites with similar vegetation and of similar altitude indicating that coupling may be mediated through shared environmental stimuli. When coupling is indicated, convergence is equally likely to occur on a peak state as a trough, indicating that synchronization may be brought about by the response of populations to a combination of different stimuli rather than by the action of any single process. PMID- 12639327 TI - A retrospective analysis of the clinical case records of 'autistic psychopaths' diagnosed by Hans Asperger and his team at the University Children's Hospital, Vienna. AB - To date, it is questionable whether the diagnostic criteria for Asperger syndrome (AS) as stated by ICD-10 or DSM-IV still reflect Asperger's original account of 'autistic psychopathy' (AP) from the 1940s. The present study examined 74 clinical case records of children with AP diagnosed by Hans Asperger and his team at the Viennese Children's Clinic and Asperger's private practice between 1950 and 1986. The characteristic features of the children are outlined, including reasons for referral, parental background, behavioural problems, cognitive functioning, communication and interests. Results show that the patients of Asperger described in our study represent a subgroup of children with very high intellectual functioning, specific circumscribed interests and talents but impaired social, communication and motor skills. Sixty-eight percent of the sample met ICD-10 criteria for AS, while 25% fulfilled the diagnostic criteria for autism. Implications for the diagnosis of AS are discussed. PMID- 12639326 TI - Understanding autism: insights from mind and brain. AB - Autism is a developmental disorder characterized by impaired social interaction and communication as well as repetitive behaviours and restricted interests. The consequences of this disorder for everyday life adaptation are extremely variable. The general public is now more aware of the high prevalence of this lifelong disorder, with ca. 0.6% of the population being affected. However, the signs and symptoms of autism are still puzzling. Since a biological basis of autism was accepted, approaches from developmental cognitive neuroscience have been applied to further our understanding of the autism spectrum. The study of the behavioural and underlying cognitive deficits in autism has advanced ahead of the study of the underlying brain abnormalities and of the putative genetic mechanisms. However, advances in these fields are expected as methodological difficulties are overcome. In this paper, recent developments in the field of autism are outlined. In particular, we review the findings of the three main neuro-cognitive theories of autism: theory-of-mind deficit, weak central coherence and executive dysfunction. PMID- 12639328 TI - Identifying neurocognitive phenotypes in autism. AB - Autism is a complex disorder that is heterogeneous both in its phenotypic expression and its etiology. The search for genes associated with autism and the neurobiological mechanisms that underlie its behavioural symptoms has been hampered by this heterogeneity. Recent studies indicate that within autism, there may be distinct subgroups that can be defined based on differences in neurocognitive profiles. This paper presents evidence for two kinds of subtypes in autism that are defined on the basis of language profiles and on the basis of cognitive profiles. The implications for genetic and neurobiological studies of these subgroups are discussed, with special reference to evidence relating these cognitive phenotypes to volumetric studies of brain size and organization in autism. PMID- 12639330 TI - Does the perception of moving eyes trigger reflexive visual orienting in autism? AB - Does movement of the eyes in one or another direction function as an automatic attentional cue to a location of interest? Two experiments explored the directional movement of the eyes in a full face for speed of detection of an aftercoming location target in young people with autism and in control participants. Our aim was to investigate whether a low-level perceptual impairment underlies the delay in gaze following characteristic of autism. The participants' task was to detect a target appearing on the left or right of the screen either 100 ms or 800 ms after a face cue appeared with eyes averting to the left or right. Despite instructions to ignore eye-movement in the face cue, people with autism and control adolescents were quicker to detect targets that had been preceded by an eye movement cue congruent with target location compared with targets preceded by an incongruent eye movement cue. The attention shifts are thought to be reflexive because the cue was to be ignored, and because the effect was found even when cue-target duration was short (100 ms). Because (experiment two) the effect persisted even when the face was inverted, it would seem that the direction of movement of eyes can provide a powerful (involuntary) cue to a location. PMID- 12639329 TI - Why is joint attention a pivotal skill in autism? AB - Joint attention abilities play a crucial role in the development of autism. Impairments in joint attention are among the earliest signs of the disorder and joint attention skills relate to outcome, both in the 'natural course' of autism and through being targeted in early intervention programmes. In the current study, concurrent and longitudinal associations between joint attention and other social communication abilities measured in a sample of infants with autism and related pervasive developmental disorders at age 20 months, and language and symptom severity at age 42 months, were examined. Extending the findings from previous studies, joint attention ability was positively associated with language gains and (lower) social and communication symptoms, and imitation ability was also positively associated with later language. Some specificity in the association between different aspects of joint attention behaviours and outcome was found: declarative, triadic gaze switching predicted language and symptom severity but imperative, dyadic eye contact behaviours did not. Further, although joint attention was associated with later social and language symptoms it was unrelated to repetitive and stereotyped symptoms, suggesting the latter may have a separate developmental trajectory. Possible deficits in psychological and neurological processes that might underlie the impaired development of joint attention in autism are discussed. PMID- 12639331 TI - The pathogenesis of autism: insights from congenital blindness. AB - There is substantial heterogeneity in the aetiology and clinical presentation of autism. So how do we account for homogeneity in the syndrome? The answer to this question will be critical for any attempt to trace the links between brain pathology and the psychological disabilities that characterize autism. One possibility is that the source of homogeneity in autism is not to be found 'in the child', but rather in dysfunction of the system constituted by child-in relation-to-other. We have been exploring this hypothesis through the study of congenitally blind children, among whom features of autism, and the syndrome of autism itself, are strikingly common. To justify such an approach, one needs to establish that the clinical features in blind children have qualities that are indeed 'autistic-like'. We conducted systematic observations of the social interactions of two matched groups of congenitally blind children who do not have autism, rating their social engagement, emotional tone, play and language during three sessions of free play in the school playground. The qualities of social impairment in the more disabled children were similar to those in sighted children with autism. Additional evidence came from independent ratings of the children in a different play setting: on the childhood autism rating scale (CARS), the socially impaired children had 'autistic-like' abnormalities in both social and non-social domains. If we can determine the way in which congenital blindness predisposes to features of autism, we shall be in a better position to trace the developmental pathways that lead to the syndrome in sighted children. PMID- 12639333 TI - The systemizing quotient: an investigation of adults with Asperger syndrome or high-functioning autism, and normal sex differences. AB - Systemizing is the drive to analyse systems or construct systems. A recent model of psychological sex differences suggests that this is a major dimension in which the sexes differ, with males being more drawn to systemize than females. Currently, there are no self-report measures to assess this important dimension. A second major dimension of sex differences is empathizing (the drive to identify mental states and respond to these with an appropriate emotion). Previous studies find females score higher on empathy measures. We report a new self-report questionnaire, the Systemizing Quotient (SQ), for use with adults of normal intelligence. It contains 40 systemizing items and 20 control items. On each systemizing item, a person can score 2, 1 or 0, so the SQ has a maximum score of 80 and a minimum of zero. In Study 1, we measured the SQ of n = 278 adults (114 males, 164 females) from a general population, to test for predicted sex differences (male superiority) in systemizing. All subjects were also given the Empathy Quotient (EQ) to test if previous reports of female superiority would be replicated. In Study 2 we employed the SQ and the EQ with n = 47 adults (33 males, 14 females) with Asperger syndrome (AS) or high-functioning autism (HFA), who are predicted to be either normal or superior at systemizing, but impaired at empathizing. Their scores were compared with n = 47 matched adults from the general population in Study 1. In Study 1, as predicted, normal adult males scored significantly higher than females on the SQ and significantly lower on the EQ. In Study 2, again as predicted, adults with AS/HFA scored significantly higher on the SQ than matched controls, and significantly lower on the EQ than matched controls. The SQ reveals both a sex difference in systemizing in the general population and an unusually strong drive to systemize in AS/HFA. These results are discussed in relation to two linked theories: the 'empathizing systemizing' (E-S) theory of sex differences and the extreme male brain (EMB) theory of autism. PMID- 12639332 TI - The enactive mind, or from actions to cognition: lessons from autism. AB - Normative-IQ individuals with autism are capable of solving explicit social cognitive problems at a level that is not matched by their ability to meet the demands of everyday social situations. The magnitude of this discrepancy is now being documented through newer techniques such as eye tracking, which allows us to see and measure how individuals with autism search for meaning when presented with naturalistic social scenes. This paper offers an approach to social cognitive development intended to address the above discrepancy, which is considered a key element for any understanding of the pathophysiology of autism. This approach, called the enactive mind (EM), originates from the emerging work on 'embodied cognitive science', a neuroscience framework that views cognition as bodily experiences accrued as a result of an organism's adaptive actions upon salient aspects of the surrounding environment. The EM approach offers a developmental hypothesis of autism in which the process of acquisition of embodied social cognition is derailed early on, as a result of reduced salience of social stimuli and concomitant enactment of socially irrelevant aspects of the environment. PMID- 12639334 TI - Towards an understanding of the mechanisms of weak central coherence effects: experiments in visual configural learning and auditory perception. AB - The weak central coherence hypothesis of Frith is one of the most prominent theories concerning the abnormal performance of individuals with autism on tasks that involve local and global processing. Individuals with autism often outperform matched nonautistic individuals on tasks in which success depends upon processing of local features, and underperform on tasks that require global processing. We review those studies that have been unable to identify the locus of the mechanisms that may be responsible for weak central coherence effects and those that show that local processing is enhanced in autism but not at the expense of global processing. In the light of these studies, we propose that the mechanisms which can give rise to 'weak central coherence' effects may be perceptual. More specifically, we propose that perception operates to enhance the representation of individual perceptual features but that this does not impact adversely on representations that involve integration of features. This proposal was supported in the two experiments we report on configural and feature discrimination learning in high-functioning children with autism. We also examined processes of perception directly, in an auditory filtering task which measured the width of auditory filters in individuals with autism and found that the width of auditory filters in autism were abnormally broad. We consider the implications of these findings for perceptual theories of the mechanisms underpinning weak central coherence effects. PMID- 12639335 TI - Disentangling weak coherence and executive dysfunction: planning drawing in autism and attention-deficit/hyperactivity disorder. AB - A tendency to focus on details at the expense of configural information, 'weak coherence', has been proposed as a cognitive style in autism. In the present study we tested whether weak coherence might be the result of executive dysfunction, by testing clinical groups known to show deficits on tests of executive control. Boys with autism spectrum disorders (ASD) were compared with age- and intelligence quotient (IQ)-matched boys with attention deficit/hyperactivity disorder (ADHD), and typically developing (TD) boys, on a drawing task requiring planning for the inclusion of a new element. Weak coherence was measured through analysis of drawing style. In line with the predictions made, the ASD group was more detail-focused in their drawings than were either ADHD or TD boys. The ASD and ADHD groups both showed planning impairments, which were more severe in the former group. Poor planning did not, however, predict detail-focus, and scores on the two aspects of the task were unrelated in the clinical groups. These findings indicate that weak coherence may indeed be a cognitive style specific to autism and unrelated to cognitive deficits in frontal functions. PMID- 12639336 TI - The reach-to-grasp movement in children with autism spectrum disorder. AB - Autism is associated with a wide and complex array of neurobehavioural symptoms. Examination of the motor system offers a particularly appealing method for studying autism by providing information about this syndrome that is relatively immune to experimental influence. In this article, we considered the relationship between possible movement disturbance and symptoms of autism and introduced an experimental model that may be useful for rehabilitation and diagnostic purposes: the reach-to-grasp movement. Research is reviewed that characterizes kinematically the reach-to-grasp movement in children with autism compared with age-matched 'controls'. Unlike the age-matched children, autistic children showed differences in movement planning and execution, supporting the view that movement disturbances may play a part in the phenomenon of autism. PMID- 12639337 TI - Investigating individual differences in brain abnormalities in autism. AB - Autism is a psychiatric syndrome characterized by impairments in three domains: social interaction, communication, and restricted and repetitive behaviours and interests. Recent findings implicate the amygdala in the neurobiology of autism. In this paper, we report the results of a series of novel experimental investigations focusing on the structure and function of the amygdala in a group of children with autism. The first section attempts to determine if abnormality of the amygdala can be identified in an individual using magnetic resonance imaging in vivo. Using single-case voxel-based morphometric analyses, abnormality in the amygdala was detected in half the children with autism. Abnormalities in other regions were also found. In the second section, emotional modulation of the startle response was investigated in the group of autistic children. Surprisingly, there were no significant differences between the patterns of emotional modulation of the startle response in the autistic group compared with the controls. PMID- 12639352 TI - Laparoscopic radical nephrectomy and nephroureterectomy. PMID- 12639338 TI - The role of the fusiform face area in social cognition: implications for the pathobiology of autism. AB - A region in the lateral aspect of the fusiform gyrus (FG) is more engaged by human faces than any other category of image. It has come to be known as the 'fusiform face area' (FFA). The origin and extent of this specialization is currently a topic of great interest and debate. This is of special relevance to autism, because recent studies have shown that the FFA is hypoactive to faces in this disorder. In two linked functional magnetic resonance imaging (fMRI) studies of healthy young adults, we show here that the FFA is engaged by a social attribution task (SAT) involving perception of human-like interactions among three simple geometric shapes. The amygdala, temporal pole, medial prefrontal cortex, inferolateral frontal cortex and superior temporal sulci were also significantly engaged. Activation of the FFA to a task without faces challenges the received view that the FFA is restricted in its activities to the perception of faces. We speculate that abstract semantic information associated with faces is encoded in the FG region and retrieved for social computations. From this perspective, the literature on hypoactivation of the FFA in autism may be interpreted as a reflection of a core social cognitive mechanism underlying the disorder. PMID- 12639353 TI - Four analgesic techniques for shockwave lithotripsy: eutectic mixture local anesthetic is a good alternative. AB - BACKGROUND AND PURPOSE: Various sedative and analgesic medication has been used for shockwave lithotripsy (SWL). The aim of this study was to evaluate the efficacy of different anesthesia modalities in these patients. PATIENTS AND METHODS: One hundred patients were randomly divided into four groups. The first (Group F) received fentanyl 1 microg/kg intravenously (IV), the second (Group D) received diclofenac sodium 1 mg/kg intramuscularly (IM), the third (Group T) received tramadol 1.5 mg/kg IM, and the fourth (Group E) was given 15 g of eutectic mixture local anesthetic (EMLA) cream containing lidocaine and prilocaine. After routine preoperative evaluation, all patients received midazolam 2 mg IV 5 minutes before lithotripsy for sedative premedication. In all groups, a supplemental 25-microg bolus of fentanyl was administered IV when patients complained of pain, moved, or grimaced in response to the shockwaves. Pain intensity was evaluated on a 0- to 100-mm visual analog scale (VAS). The level of sedation was determined using the Observer's Assessment of Alertness/Sedation (OAS/S). Side effects such as bradypnea, oxygen desaturation, bradycardia, pruritus, and nausea and vomiting were recorded. RESULTS: There were no statistically significant differences among the four groups with regard to VAS, OAS/S scores, or side effects. In Group F, the mean arterial pressure was decreased significantly at 10 and 20 minutes. The patients in this group also manifested a decrease of oxygen saturation at the first, tenth, and twentieth minutes and the end of SWL. CONCLUSION: Application of EMLA cream was as safe and effective as fentanyl, diclofenac, and tramadol, and reduction of the fentanyl dose during SWL was possible. PMID- 12639354 TI - Erect and supine radiographs to assess effectiveness of SWL for stones in a caliceal diverticulum or dilated calix. AB - BACKGROUND AND PURPOSE: There are patients who have been treated with shockwave lithotripsy (SWL) for stones in a caliceal diverticulum (CD) or a dilated calix in whom the degree of fragmentation is difficult to assess. The aim of this study was to see if adequate fragmentation could be confirmed by the demonstration of layering of fine fragments on an erect radiograph. PATIENTS AND METHODS: Over a period of 9 months, 13 patients with stones in a CD or a dilated calix with a stenosed infundibulum were studied 2 weeks after SWL with erect and supine radiographs. RESULTS: One of eight patients with a CD cleared all fragments. Two patients showed no change in the appearance of the stone on either the supine or the erect film, and of the remaining five patients, three demonstrated layering of sand at the base of the CD. Five patients with a dilated calix and a narrow infundibulum were also studied: two of these patients became stone free, two demonstrated complete fragmentation with layering of the sand, and in one patient, a 4-mm fragment, hidden within the sand, was revealed only on the erect film. CONCLUSION: There are patients who have been treated with SWL for stones in a CD or a dilated calix in whom adequate fragmentation is difficult to demonstrate. An erect radiograph in these patients may demonstrate layering of the fragments to confirm fragmentation and obviate repeat SWL. PMID- 12639355 TI - Laparoscopic adrenal surgery in a Brazilian center. AB - BACKGROUND AND PURPOSE: The laparoscopic approach to the adrenal gland was first reported in 1992. Since then, many publications about this issue have come from Europe, Japan, and North America. We reviewed our 7-year experience with laparoscopic adrenal surgery. This is the first large series presented from Latin America. PATIENTS AND METHODS: Laparoscopic adrenalectomy was carried out in 61 female and 33 male patients between January 1994 and February 2001. Their ages ranged from 1 to 72 (42.8 +/- 16.4 years) years. Ten patients (10.6%) were 20 years or younger, 10 (10.6%) had unilateral tumors >4 cm, 22 (23.4%) had a Body Mass Index >/= 30, and 10 (10.6%) had had previous open upper abdominal surgery. The size of the lesion ranged from 1 to 9 cm (2.9 +/- 4 cm). Ninety-seven operations were performed, of which 91 were unilateral and 6 were bilateral, adding up to 103 adrenalectomies. Among the 97 procedures, the lateral transperitoneal approach was employed in 94 cases, whereas a lateral retroperitoneal approach enabled 3 adrenalectomies. RESULTS: Unilateral procedures lasted 117 +/- 43.7 minutes (range 45-250 minutes); bilateral procedures lasted 186 +/- 91.6 minutes (range 100-345 minutes). Five operations (5.1%) were converted to open surgery. Twenty patients (21.3%) suffered complications, 8 (8.5%) being intraoperative and 12 (12.8%) postoperative. Six cases (6.4%) were considered major complications. No deaths occurred. The blood transfusion rate was 4.2%. The hospital stay averaged 6.1 +/- 15.3 days (range 1 140 days). The follow-up period was 17 +/- 12.6 months (range 1-60 months). CONCLUSIONS: Laparoscopic adrenalectomy is feasible and has excellent results in properly selected patients. PMID- 12639356 TI - Transplant ureter stricture: Acucise endoureterotomy and balloon dilation are effective. AB - PATIENTS AND METHODS: Eight patients with ureteral stricture after renal transplantation underwent minimally invasive treatment with Acucise incision or balloon dilation. Acucise endoureterotomy was used to treat four patients with strictures at the ureterovesical anastomosis, and balloon dilation was used to treat four patients with a ureteroureterostomy stricture. Success was defined as an acceptable serum creatinine concentration in the absence of hydronephrosis with at least 1 year of follow-up. RESULTS: Acucise endoureterotomy for ureterovesical anastomosis stricture was successful in two of three patients (67%) with a mean follow-up of 20 months. One patient had an indeterminate outcome. Balloon dilation of strictured ureteroureterostomy was successful in three of four patients (75%) with a mean follow-up of 23.7 months. Three of the four patients with previously failed open revision were treated successfully with endourologic techniques. The two patients in whom treatment failed had strictures >/=1.5 cm and manifested comorbidities including diabetes mellitus. CONCLUSION: As our results are comparable to those of other published series, endourologic management of transplant ureteral stenosis is a reasonable strategy. PMID- 12639357 TI - Creation of ureteropelvic junction obstruction and its correction by chemical glue-assisted laparoscopic dismembered pyeloplasty. AB - PURPOSE: We established a porcine model of ureteropelvic junction (UPJ) obstruction using a laparoscopic technique and assessed the outcome of standard suture-assisted and chemical glue-assisted laparoscopic pyeloplasty. MATERIALS AND METHODS: Female domestic pigs (N = 20) underwent laparoscopic suture-ligature to create UPJ obstruction. One month later, laparoscopic end-to-end anastomosis was performed to correct the obstruction: with standard suturing techniques in 10 animals and with chemical (cyanoacrylate) glue in the other 10. Postoperative ureteral stents were not used. Four weeks postoperatively, intravenous urography was performed to evaluate the patency of the anastomoses. The UPJ was procured by laparotomy to assess the anastomoses and periureteral fibrosis histologically. RESULTS: The UPJ obstruction was created in an average of 15 +/- 6 minutes. There was no early postoperative mortality. Eighteen pigs survived for at least 1 month, and UPJ obstruction developed in 17 (95%). Microscopically, the lumen of the UPJ was partially occluded, measuring an average of 40% +/- 5% of normal. After laparoscopic correction, a patent UPJ was found in seven of nine animals treated with traditional sutures. Among the eight animals with chemically glued anastomoses, none had a patent UPJ, and severe periureteral adhesions and intraluminal fibrosis were noted at the pyeloplasty site. Marked ureteral tortuosity was present in six of the eight pigs receiving glue-assisted pyeloplasty but in none of the animals having suture-assisted pyeloplasty. CONCLUSIONS: Ureteropelvic junction obstruction was established by laparoscopic suture-ligature in a porcine model with a 95% success rate. Chemical glue assisted anastomosis was inferior to standard laparoscopic sutures for pyeloplasty to correct the obstruction. PMID- 12639359 TI - Spontaneous rupture of the ureter after cystectomy and creation of orthotopic ileal neobladder: treatment with percutaneous nephrostomy and drainage. AB - A case of rupture of the upper ureter following creation of a continent orthotopic neobladder, treated conservatively with percutaneous nephrostomy and drainage of a paranephric collection, is reported. PMID- 12639358 TI - Controversial cases in endourology. PMID- 12639360 TI - Effects of cryoablation on short-term development of urinary fistulas in the porcine kidney. AB - PURPOSE: To determine the effect of cryoablation on the collecting system in the porcine kidney and the possible development of pelvic injury or fistula. MATERIALS AND METHODS: Six kidneys from three domestic swine were utilized. Under MRI guidance, a 3-mm cryoprobe was placed percutaneously and advanced to a point adjacent to the collecting system. The tissue then was frozen to -40 degrees C for 4 to 6 minutes to create a 2-cm iceball. At 1 week postprocedure, the kidneys were harvested and underwent gross and microscopic examination for evidence of fistula formation or renal pelvis injury. RESULTS: The procedures were performed without complications, although during placement of the probe in one pig, the collecting system was inadvertently entered. The diameter of the iceballs ranged from 2.0 to 2.5 cm. On pathologic examination, five of six of the kidneys, excluding the one noted above, were found to have no gross evidence of injury or fistula formation from the collecting system. Microscopic examination of the collecting system demonstrated necrosis extending up to the mucosa but no injury to the collecting system itself. CONCLUSIONS: In this animal model, no evidence of damage to the renal collecting system secondary to freezing was found. Treatment of renal tumors that are close to the collecting system may be possible without damage to the collecting system. PMID- 12639361 TI - Objective and subjective changes in patients with Peyronie's disease after management with shockwave therapy. AB - BACKGROUND AND PURPOSE: Very few reports have been published on the management of Peyronie's disease by shockwave therapy. Existing publications on this topic are based on subjective improvement described by the patients themselves. Our aim was to determine objectively the effect of shockwave therapy on the signs and symptoms caused by Peyronie's disease. PATIENTS AND METHODS: To date, 65 patients (age 58.4 +/- 8.7 years) have been enrolled in a therapeutic pilot study. The disease duration was 33.7 +/- 42.9 months. Inclusion criteria were palpable plaque together with deviation, pain (visual pain scale), or loss of distal rigidity. Clinical examination (prior to first therapy and 1, 6, and 18 months after last shockwave delivery) included palpation and sonography of the plaque (mean surface size 2.2 +/- 1.1 cm(2)), measurement of deviation, assessment of pain and distal loss of rigidity (artificial erection induced by intracavernosal injection of 5 microg of alprostadil [Caverject]). Shockwaves (1,000 impulses at 12 kV per square centimeter of plaque) were delivered to the nonerect penis once a week for a period of 5 weeks with the Minilith; Storz Medical. RESULTS: Eighteen months (N = 35) after the last shockwave session, the deviation angle had decreased from 59.3 degrees +/- 38.1 degrees to 49.3 degrees +/- 32.5 degrees (N = 24; P = 0.1496). Pain during erection disappeared in 15 of 17 patients and was reduced in 1 other patient (P < 0.0001). There was no effect on distal rigidity in any patient. Six patients achieved satisfactory sexual intercourse (vaginal penetration) before and 15 patients after shockwave therapy. The adverse effects were small skin hematomas in 90% of patients and initial transient macrohematuria in 30%. CONCLUSION: Our study demonstrates objective and subjective changes in patients with Peyronie's disease after shockwave therapy. Artificial erection served as a control to assess improvement of the deviation angle. PMID- 12639363 TI - Health-related quality of life among women of reproductive age: Behavioral Risk Factor Surveillance System (BRFSS), 1998-2001. PMID- 12639364 TI - Toward optimal health: the experts discuss gastrointestinal dysfunction. PMID- 12639365 TI - Current medical, rehabilitation, and surgical management of vertebral compression fractures. AB - Approximately 25% of women over the age of 50 in the United States will suffer one or more vertebral compression fractures (VCFs) related to osteoporosis. VCFs are the most common of all osteoporotic fractures, with an incidence of approximately 700,000 annually. Such a fracture may cause significant pain, disability, and loss of general health and mobility and may lead to a progressive decline in quality of life. This is a review of the clinical literature on VCFs, including patient presentation, methods of diagnosis, and current rehabilitation and medical management. Much of the pain and disability that follow a VCF may be minimized by addressing the psychological impact of such a fracture, using current medications to help limit bone loss and preserve bone mass, adhering to a well-planned rehabilitation program, and consideration of vertebroplasty or kyphoplasty in appropriate patients. A multifaceted approach will help to optimize recovery from a VCF related to osteoporosis. PMID- 12639366 TI - Thyroid hormone use and the risk of hip fracture in women > or = 65 years: a case control study. AB - BACKGROUND: There is controversy about whether thyroid hormone therapy may lead to osteoporosis, and less is known about the clinically more important end point of whether its use increases fracture risk. METHODS: We used a case-control study to examine the association between thyroid hormone use and hip fractures among older women in a large managed care organization in Northern California. The subjects were 501 women > or =65 years of age who were hospitalized for hip fractures and 533 age-matched controls without hip fractures. RESULTS: No difference in the ever use or duration of use of exogenous thyroid hormone was found between cases and controls (odds ratio [OR] 1.1, 95% confidence interval [CI] 0.8,1.6). Hip fracture was associated with evidence of visual impairment, prior use of steroids, and number of falls. CONCLUSIONS: In women > or =65 years, an independent effect of thyroid hormone use on the risk of hip fracture was not found. This finding is reassuring, given the large number of women on thyroid hormone therapy today. PMID- 12639367 TI - Failure of tibolone to potentiate the pharmacological effect of fluoxetine in postmenopausal major depression. AB - BACKGROUND: Perimenopausal depression has been attributed to physiological progressive estrogen decline. Estrogen and derivatives have some mood-enhancing effects, although studies of using estrogen as an antidepressant have had mixed results. The gonadomimetic drug tibolone stimulates estrogen receptors in a tissue-selective fashion, increasing the gonadal activity without causing some of the usual side effects of other estrogen preparations. METHODS: A total of 31 postmenopausal outpatients with a major depressive disorder (MDD) participated in the study. Sixteen received the antidepressant fluoxetine (20 mg/day) plus tibolone (2.5 mg/day), and 15 received the same dose of fluoxetine plus placebo, assigned in a randomized fashion. RESULTS: After 8 weeks of treatment, the two groups had a similar level of improvement in their depressive symptoms. Both treatments were well tolerated, without significant side effects. Pretreatment and posttreatment serum hormonal levels did not predict the final response. CONCLUSIONS: Combining tibolone and fluoxetine did not represent a more robust antidepressant response than fluoxetine alone in postmenopausal women with MDD. PMID- 12639368 TI - Sexual orientation and variation in physical and mental health status among women. AB - OBJECTIVE: To assess and compare the physical and mental health status of women of differing sexual orientation within a population-based sample. METHODS: We used a population-based telephone survey performed using random digit dialing techniques. Our study population was drawn from the 1999 Los Angeles County Health Survey and included women age 18-64 years who reported their sexual orientation (98%, n = 4135). These analyses include 4023 heterosexuals, 69 bisexuals, and 43 lesbians. RESULTS: We assessed the unique association of sexual orientation with physical and mental health status using bivariate and multivariate analyses. Both lesbians and bisexuals were more likely than heterosexual women to report a diagnosis of heart disease. Among women with a depressive disorder, lesbians were more likely than heterosexuals to be using an antidepressant medication. Compared with heterosexuals within the preceding 30 days, lesbians reported significantly more days of poor mental health, and bisexuals reported significantly more days of poor physical health. However, there were no significant differences by sexual orientation in impaired ability to perform daily activities due to physical or mental health. CONCLUSIONS: In this rare opportunity to use population-based data to study lesbian and bisexual health, we found that sexual orientation as a nonheterosexual woman was associated with increased rates of poor physical and mental health. We believe these findings support the need for the increased systematic study of the relationship between sexual orientation and health. PMID- 12639369 TI - Gender and noninvasive diagnosis of coronary artery disease in women and men. AB - OBJECTIVES: The purpose of this prospective study was to investigate the impact of the gender of physicians and patients on the accuracy of diagnosing coronary artery disease (CAD) based on the patient history only. METHODS: Screening involved 1082 consecutive patients undergoing coronary angiography for suspected CAD. Known CAD was an exclusion criterion. The inclusion criteria were met by 144 patients (47 females). All patients were inverviewed by both an experienced male cardiologist (n=6) and an experienced female cardiologist (n=5), who had to state before coronary angiography if CAD was present relying solely on the patient interview. Eligible subjects also answered a questionnaire. Diagnostic performance to assess the presence or absence of CAD by history taking by male and female cardiologists, exercise testing, patient self-assessment, and the questionnaire were compared. RESULTS: Coronary angiography showed significant CAD in 20 females (43%) and 70 males (72%). Diagnostic accuracy was not different between male (79%) and female (79%) cardiologists and comparable to the results of exercise testing (74%) but better than self-assessment by the patients (65%, p=0.01) or a questionnaire (68%, p=0.01). The accuracy of female physicians was better in men than in women (85% vs. 66%, p=0.01). The specificity of male physicians tended to be better in women than in men (74% vs. 59%, p=0.25). Female cardiologists overestimated the presence of CAD in women (specificity 48% vs. 74%, p=0.04). CONCLUSIONS: The female cardiologists did not assess women more accurately than did their male colleagues. The diagnostic accuracy of these male and female cardiologists was quite good. However, both tend to assess patients of the opposite gender more specifically. There may be an impact of the gender of the physician on the accuracy of diagnosis of CAD by history taking. PMID- 12639370 TI - Provider characteristics and mammography recommendation among women in their 40s and 50s. AB - OBJECTIVES: Healthcare provider recommendation for mammography is one of the strongest predictors of women's mammography use, but few studies have examined the association of provider characteristics with mammography recommendations. We examined the relationship of provider gender, age, medical specialty, and duration of relationship with the patient to report mammography recommendation. METHODS: Participants were women ages 40-45 and 50-55 who were part of a larger intervention study of decision making about mammography. We examined the relationship of provider characteristics to patient-reported mammography recommendations at baseline and at 24-month follow-up. RESULTS: At baseline, 74% of women in their 40s and 79% of women in their 50s reported provider mammography recommendations within the prior 2 years. Proportions were similar at the 24 month follow-up. In multivariate logistic regression models including both patient and provider characteristics, women in their 40s who had female providers were more likely to report mammography recommendations than those with male providers at baseline (OR=1.83, p=0.01) and follow-up (OR=1.74, p=0.03). Among women in their 50s, participants whose regular providers were primary care physicians were more likely to report recommendations at baseline than those whose regular providers were obstetrician/gynecologists (OR=1.68, p=0.03). CONCLUSIONS: About one fourth of women in this study reported not having been advised by a healthcare provider to have a mammogram. All women in the study had health insurance. Among women in their 40s, for whom mammography guidelines were controversial at the time of data collection, provider gender was an important predictor of patient-reported mammography recommendation. PMID- 12639371 TI - Physician gender, patient gender, and primary care. AB - BACKGROUND: Studies of the effects of physician gender on patient care have been limited by selected samples, examining a narrow spectrum of care, or not controlling for important confounders. We sought to examine the role of physician and patient gender across the spectrum of primary care in a nationally representative sample, large enough to examine the role of gender concordance and adjust for confounding variables. METHODS: We examined the relationships between physician and patient gender using nationally representative samples (the U. S. National Ambulatory Medical Care Surveys from 1985 to 1992) of encounters of 41,292 adult patients with 1470 primary care physicians (internists, family physicians, and obstetrician/gynecologists). Factors examined included physician (age, gender, region, rural location), patient (age, gender, race, insurance), and visit characteristics (diagnoses, gender-specific and nonspecific prevention, duration, continuity, and disposition). RESULTS: After multivariate adjustment, female physicians were more likely to see female patients, had longer visit durations, and were more likely to perform female prevention procedures and make some follow-up arrangements and referrals. Female physicians were slightly more likely to check patients blood pressure, but there were no significant differences in other nongender-specific prevention procedures or use of psychiatric diagnoses. Among encounters without breast or pelvic examinations, visit length was not related to physician gender, but length was longer in gender concordant visits than gender-discordant visits. CONCLUSIONS: Female physicians were more likely to deliver female prevention procedures, but few other physician gender differences in primary care were observed. Physician-patient gender concordance was a key determinant of encounters. PMID- 12639380 TI - Synthesis and self-assembly of colloidal nanoparticles. AB - We describe recent developments in the synthesis of semiconductor nanoparticles, which lead to a substantial improvement of the luminescence quantum efficiency. Concerning a theoretical model for the growth of an ensemble of nanoparticles, the highest quantum efficiencies are achieved in particles that grow under conditions of a rapid exchange of monomers at the particle surface, leading to a smooth surface structure. Selective etching, core-shell formation and doping of nanoparticles are also discussed as fluorescence-enhancing preparative techniques. Examples of self-assembly of almost-uniformly-sized nanoparticles are given, which result in two-dimensional and three-dimensional superlattices, colloidal crystals and crystalline structures built-up from particles of different sizes. Finally, the self-assembled oriented attachment of quasi spherical ZnO nanoparticles onto single-crystalline nanorods is presented. PMID- 12639372 TI - Pulmonary lymphangioleiomyomatosis (LAM): examining oral contraceptive pills and the onset of disease. AB - BACKGROUND: Lymphangioleiomyomatosis (LAM) is a rare pulmonary disease of women of unknown etiology. Diffuse cystic changes associated with multifocal clusters of smooth muscle-like cells (LAM cells) replace normal lung parenchyma, leading to progressive loss of lung function. Reports of exacerbations of LAM with the use of exogenous estrogens and during pregnancy suggest an association between hormones and clinical deterioration. OBJECTIVES: To conduct the largest case series to date in order to characterize the demographics of a living cohort of patients with LAM. To evaluate the effect of oral contraceptive pill (OCPs) use on the age at onset of symptoms in a large population of women with LAM. METHODS: The study population was drawn from the University of Miami (UM) LAM database consisting of 91 living women previously diagnosed with LAM by a physician. A self-administered questionnaire was mailed to the participants. RESULTS: The final response rate was 91.2% (83 of 91), and the study participants were 96.4% white, 1.2% Asian, 1.2% African American, and 1.2% American Indian; 68.4% identified their ethnicity as European and 14.5% as Jewish. Of the women surveyed, 73 were still menstruating at the time of LAM diagnosis. Among these women, the rate of use of OCPs was approximately 27%, similar to the rate in the general population. There was a significant difference between the age at onset of symptoms in women using OCPs vs. women not using OCPs (29.2 +/- 4.7 vs. 32.9 +/- 8.0, p = 0.0397). CONCLUSIONS: The present study suggests that OCPs may serve as catalysts to promote an earlier occurrence of LAM. PMID- 12639381 TI - Shape control and applications of nanocrystals. AB - Inorganic nanocrystals with well-defined shapes are important for understanding basic size-dependent scaling laws, and may be useful in a wide range of applications. Methods for controlling the shapes of inorganic nanocrystals are evolving rapidly. This paper will focus on how we currently control the shape of nanocrystals and this will be illustrated using CdSe and Co nanocrystals as examples for semiconductors and for metals. These materials show a more pronounced variation of fundamental properties with aspect ratio. However, to take advantage of these shape-dependent properties in possible applications, several challenges need to be overcome. Issues such as alignment, high quantum yield and photostability and precise control of three-dimensional structures need to be addressed. These challenges, as well as several potential applications, will be described briefly. PMID- 12639382 TI - Confinement effects and tunnelling through quantum dots. AB - Several recent theoretical advances concerning semiconductor quantum dots are reviewed. First of all, the effect of the quantum confinement on the energy gap is revisited on the basis of GW and Bethe-Salpeter calculations, showing that the excitonic gap is practically equal to the ordinary eigenvalue gap of single particle approximations. The second part demonstrates that it is now possible to calculate the conductance peaks for the tunnelling current through a nanostructure. Finally, we discuss in some detail the concept of a macroscopic dielectric constant for nanostructures, showing that, except for a thin surface layer, the local dielectric constant still keeps its bulk value down to pretty small nanostructures. PMID- 12639383 TI - Quantum dots as dynamical systems. AB - Quantum dots show a range of time-dependent behaviours. We show that the polarity of II-VI nanoparticles has important dynamical implications for electronic, vibrational and other phenomena. Polarity-dependent phenomena are found even for nearly spherical stoichiometric clusters of ZnO and ZnS in studies based on interatomic potentials or on a plane-wave density-functional approach. We find a substantial dipole moment for free nanoparticles, whether of the zinc blende or wurtzite structure. This dipole causes a highly non-uniform spin-density distribution on electronic excitation or after a change in the dot's electronic charge state. The spin density of the triplet exciton shows that the dipole aligns so as to reduce the dipole moment in the electronically excited state. The polarity of II-VI dots also affects their vibrational properties. High- and low frequency tails of the vibration density of states arise from modes strongly localized at surface atoms, near the poles of the dipole. These features, first noted for free clusters, also hold for particles embedded in a wide-gap dielectric, a-SiO(2). We present the results of molecular dynamics of the ZnS particle embedded into the silica glass, and consider the role played by the soft modes in energy-dissipation processes such as dephasing during non-radiative recombination of excitons, and energy transfer from the dot to the matrix. PMID- 12639384 TI - Dynamical behaviour of nanocrystals in transmission electron microscopy: size, temperature or irradiation effects. AB - High-resolution transmission electron microscopy shows that metal nanoparticles sinter within a fraction of a second under an electron beam at 'room temperature' as long as classical models of thermal equilibrium apply. Images exhibit crystal planes that change in orientation with time as if the particle was undergoing melting and resolidification processes. We explore whether these dynamical effects are the result of heating or transformation effects in the electron microscope or quantum fluctuations in small systems. PMID- 12639385 TI - New synthetic routes for quantum dots. AB - Various methods for the synthesis of quantum dots of compound semiconductors are discussed. Emphasis is placed on methods involving the decomposition of chalcogenide-containing precursors in tri-n-octylphosphine oxide. The use of simple coordination complexes can avoid the use of pyrophoric precursors. Approaches based on melts or the use of imino-bisdichalcogeno-diphosphinates are also discussed. These routes may be suitable for the production of large quantities of quantum dots. PMID- 12639386 TI - Selective nucleation and controlled growth: quantum dots on metal, insulator and semiconductor surfaces. AB - Nucleation and growth models are well developed for nucleation on homogeneous substrates, and they can typically be described in terms of three energy parameters. Nucleation on substrates containing point-defect traps has been investigated, at the cost of introducing more energy parameters. This paper outlines the quantitative description of such growth models, using rate and rate diffusion equations, in terms of energies for individual surface processes, with examples taken from metal-metal, metal-insulator and semiconductor growth. The challenge to modelling is to describe the large range of length and time-scales in thin-film fabrication and degradation, without relying on too many (unknown) material parameters, which often occur in combination. Separating them into elementary processes often proves to be a challenge. One typically requires selective nucleation using patterned substrates, in combination with controlled, self-organized, growth for reliable nanotechnology. Reconstructed semiconductor surfaces offer both a further challenge to modelling and an opportunity for future technology; these paradoxes are discussed briefly. PMID- 12639387 TI - Putting nanocrystals to work: from solutions to devices. AB - The colloidal route to semiconductor nanocrystals is extremely flexible, with a high degree of control over size, size distribution, surface passivation and internal structure of the nanoparticles. Simple chemically controlled techniques can be used to assemble these particles into dense films or other microscopic structures, suitable for photonic devices. Working with semiconductors or semi metals which in the bulk form have low or inverted bandgaps, and taking advantage of the blue shift in the quantum confinement regime, nanocrystals can readily be tuned to the infrared wavelengths of interest for telecommunications. Design flexibility is far greater than with conventional compound semiconductors or rare earth-doped glasses. Preliminary results demonstrating optical gain from II-VI nanocrystal films at room temperature are reported. PMID- 12639388 TI - Increasing the efficiency of erbium-based sources using silicon quantum dots. AB - Silicon nanoclusters exhibit novel and interesting optical and electrical properties that are not observed in bulk silicon. Moreover, it has been discovered that there exists a strong coupling between nanoclusters and rare earth ions that results in efficient energy exchange between the two species. This paper presents a review of recent work at University College London in this area, in which we have studied the optical properties of silicon nanoclusters in silica with rare-earth co-dopants and have developed a model for the excitation of erbium ions in erbium-doped silicon nanocrystals via coupling from optically generated excitons confined within the silicon nanoclusters. The model provides a phenomenological description of the exchange mechanism and allows us to evaluate an effective absorption cross-section for erbium that is up to four orders of magnitude higher than the corresponding value in stoichiometric silica. This paper also discusses the origin of the 1.6 eV emission band associated with the silicon nanoclusters and determines absorption cross-sections and excitonic lifetimes for nanoclusters in silica. PMID- 12639389 TI - Electrical and optical properties of semiconductor nanocrystals. AB - Application of semiconductor nanocrystals in optoelectronic devices requires an understanding not only of their emission and absorption properties, but also of the processes of charge injection and transport in nanocrystalline films. Here, we present measurements of the electrical properties of nanocrystalline films and of blends of nanocrystals with conjugated polymers. We also describe the attachment of nanocrystals to semiconductor surfaces, and we investigate the emission of nanocrystalline films in microcavity structures and at high excitation intensities. PMID- 12639390 TI - Classical versus quantum coherence. AB - Quantum dots are structures engineered to have desirable quantum-mechanical properties. Much of their potential usefulness stems from the ability to design such structures so that their passive properties can be exploited. However, there are many plans to use dots as the basis of a more active quantum engineering, in which the detailed quantum states of the dot's components are manipulated by externally imposed fields. This leads to a new requirement: our ability to model the time development of individual driven quantum systems. I shall discuss some of the problems which arise when single systems, rather than ensembles, are considered and give examples which illustrate the effects of classical and quantum incoherences. I propose a new classification of incoherent processes, based on their effect on individual wave functions. PMID- 12639391 TI - The engineering of quantum-dot devices. AB - Insofar as quantum-dot structures represent the extrapolation of today's microstructures used for electronic and optoelectronic device applications, their low-cost manufacture presents many challenges which will take some time to overcome if tunnel and quantum-well devices are any guide. PMID- 12639392 TI - Effect of lycopene on prostate LNCaP cancer cells in culture. AB - Epidemiological studies have shown an inverse relationship between serum lycopene levels and the risk of prostate cancer. The objective of this study was to measure the effect of lycopene on the proliferation of LNCaP human prostate cancer cells in culture. A new, water-dispersible lycopene in an appropriate vehicle was used. The stock solution was diluted in the medium to obtain lycopene concentrations of 10(-6), 10(-5), and 10(-4) M; their corresponding vehicles were similarly diluted to be used as controls. Cells were grown for 48 hours in RPMI 1640 medium supplemented with 10% fetal bovine serum and antibiotics. Lycopene was then added at different concentrations, and the cells were allowed to grow for 24, 48, 72, and 96 hours. Lycopene at concentrations of 10(-6) and 10(-5) M significantly reduced the growth of LNCaP cells after 48, 72, and 96 hours of incubation, by 24.4% to 42.8% (P <.05). The inhibitory effect of lycopene was significantly higher than that of the corresponding vehicle controls. In a follow up experiment, a lower range of lycopene concentrations (10(-9) to 10(-7) M) was used to determine whether there was a dose-response effect. Lycopene significantly decreased the growth of cells in a dose-dependent manner when cells were incubated for 24, 48, 72, or 96 hours (F = 3.150, 11.27, 54.51, and 297.5, respectively; P <.05). The growth inhibitory effect of lycopene on human prostate cancer cells observed in this study suggests a possibly important role for lycopene as an antioxidant in human prostate cancer; however, investigations of other mechanisms are warranted. PMID- 12639393 TI - Radioprotective role of Amaranthus gangeticus Linn.: a biochemical study on mouse brain. AB - Brain tissue is highly susceptible to oxidative damage due to its high utilization of oxygen and its poorly developed antioxidative defense mechanisms. Radiation or pro-oxidants interact with cells and tissues through secondary ionization mechanisms such as lipid peroxidation (LPO). LPO can be inhibited by antioxidants such as vitamins A, C, and E and the carotenoid beta-carotene. beta Carotene, the provitamin A, plays an important radioprotective role due to its properties as a potent free radical scavenger, singlet oxygen quencher, and lipid antioxidant. Amaranthus gangeticus Linn., widely considered as a weed, has a high content of carotene, ascorbic acid, and folate and may prove an efficient antioxidant. To evaluate the antioxidative efficacy of Amaranthus, healthy Swiss albino mice from an inbred colony were treated with alcoholic extract of A. gangeticus leaves (AE) for 2 weeks, at 800 mg/kg body weight, before radiation exposure. Irradiated mice were examined and autopsied at intervals of 1, 3, 7, 15, and 30 days after exposure. Brain was removed by skull dissection, and various biochemical changes were sought. Radiation caused a maximum increase of 27% in LPO and a maximum decrease of 27.96% in protein content at day 7 in controls. However, in the experimental group the increase in LPO was 9.98% and the increase in protein content was 18.78% at day 7. By day 30 after irradiation, AE brought these values to near-normal levels. AE protected brain biochemical activity in this murine study and may prove beneficial for clinical use as a radioprotector. PMID- 12639394 TI - Influence of Piper betle on hepatic marker enzymes and tissue antioxidant status in ethanol-treated Wistar rats. AB - Piper betle L. is a commonly used masticatory in Asia. This study was carried out to investigate the hepatoprotective and antioxidant properties of P. betle, using ethanol intoxication as a model of hepatotoxic and oxidative damage. Ethanol treated rats exhibited elevation of hepatic marker enzymes and disturbances in antioxidant defense when compared with normal rats. Oral administration of P. betle extract (100, 200, or 300 mg/kg body weight) for 30 days significantly (P <.05) decreased aspartate aminotransferase (AST), alanine aminotransferase (ALT), thiobarbituric acid reactive substances (TBARS), and lipid hydroperoxides in ethanol treated rats. The extract also improved the tissue antioxidant status by increasing the levels of nonenzymatic antioxidants (reduced glutathione, vitamin C, and vitamin E) and the activities of free radical-detoxifying enzymes such as superoxide dismutase, catalase, and glutathione peroxidase in liver and kidney of ethanol-treated rats. The highest dose of P. betle extract (300 mg/kg body weight) was most effective. The results were comparable with the known hepatoprotective drug, silymarin. These results indicate that P. betle could afford a significant hepatoprotective and antioxidant effect. PMID- 12639395 TI - Inhibition of mast cells by algae. AB - There is a history of use of algae as foods and as food additives, or nutraceuticals. Although algae are a safe component of human foods and animal feeds, the effects of the algae other than as a source of protein are not clear. We examined the prevalence of an antiinflammatory activity in selected algae using, as an assay system, the inhibition of histamine release from mast cells. Methanolic extracts of eleven algae were examined for activity to inhibit the release of histamine from mast cells in vitro. This activity was found widely among the samples tested. The activities of these extracts were not uniformly stable in acid methanol. Selected extracts studied further did not separate with the use of size-exclusion filtration filters. LH-20 chromatography suggested at least two main elution areas of activity of the Chlorella extract. In summary, we saw wide phylogenetic dispersion of mast cell inhibition activity, suggesting that this antiinflammatory property is common in algae. This effect was apparently due to multiple activities within the algal extracts. PMID- 12639396 TI - Effect of natural and synthetic antioxidants in a mouse model of chronic fatigue syndrome. AB - Chronic fatigue syndrome (CFS) is an illness characterized by persistent and relapsing fatigue, often accompanied by numerous symptoms involving various body systems. The etiology of CFS remains unclear; however, a number of studies have shown that oxidative stress may be involved in its pathogenesis. In the present study, a mouse model of CFS was used in which mice were forced to swim for one 6 minute session on each day for 15 days and the immobility period was recorded. There was a significant increase in immobility period in saline-treated mice on successive days. Intraperitoneal treatment with the potent antioxidants carvedilol (5 mg/kg) and melatonin (5 mg/kg) produced a significant reduction in immobility period. Similar results were observed with herbal preparations administered orally: Withania somnifera (100 mg/kg), quercetin (50 mg/kg), and St. John's wort (Hypericum perforatum L., 10 mg/kg). Biochemical analysis revealed that chronic swimming significantly induced lipid peroxidation and decreased glutathione (GSH) levels in the brains of mice. The rats also showed decreased levels of antioxidant defense enzymes, superoxide dismutase (SOD), and catalase. Co-administration of antioxidants carvedilol, melatonin, W. somnifera, quercetin or St. John's wort significantly reduced lipid peroxidation and restored the GSH levels decreased by chronic swimming in mice. Further, the treatment increased levels of SOD in the forebrain and of catalase. The findings strongly suggest that oxidative stress plays a significant role in the pathophysiology of CFS and that antioxidants could be useful in the treatment of CFS. PMID- 12639397 TI - Efficacy of a whey protein concentrate on the inhibition of stomach ulcerative lesions caused by ethanol ingestion. AB - The purpose of this research was to test the ability of a whey protein concentrate (WPC) to inhibit gastric mucosal ulcerative lesions caused by oral administration to rats of absolute ethanol. Acute administration (single doses) of WPC resulted in 41% inhibition of the ulcerative lesion index (ULI), and 73% inhibition was obtained with repetitive doses. In a 10-days subchronic treatment study, the inhibition was 64%, all relative to a saline treatment (negative control). Alkylation of sulfhydryl compounds by subcutaneous injection of N ethylmaleimide essentially eliminated the WPC protection. Treating the rats with an intraperitoneal injection of butathionine sulfoximine, which inhibits glutathione synthesis, reduced WPC protection to 35% and 52% for single and double doses, respectively. Taken as a whole, the results indicate that WPC does protect gastric mucosa from ethanol damage and that the protection depends on sulfhydryl compounds present in the WPC, including its capacity to stimulate glutathione synthesis. PMID- 12639398 TI - Anti-platelet fraction from Galega officinalis L. inhibits platelet aggregation. AB - A fraction from crude extract of Galega officinalis L. was purified by gel filtration on Sephadex G-25, Sepharose 4B, and ion-exchange chromatography on diethylaminoethyl (DEAE)-cellulose. The fraction with molecular weight 100-140 kDa appears to have a polysaccharide nature, including protein. The fraction inhibits platelet aggregation initiated by 25 microM adenosine 5'-diphosphate (ADP), 100 microg/ml collagen, and 0.8 U/ml thrombin with the 50% inhibiting concentration (IC(50)) being 11.2 microg/ml for ADP, and the IC(100) being 15.1 microg/ml for collagen and IC(100) 19.6 microg/ml for thrombin. PMID- 12639400 TI - In Vitro and In Vivo Lipoprotein Antioxidant Effect of a Citrus Extract and Ascorbic Acid on Normal and Hypercholesterolemic Human Subjects. AB - Polyphenols and particularly flavonoids are well known in vitro antioxidants. Their consumption in foods has been shown to decrease the risk of heart disease in epidemiological studies. Because flavonoids are consumed with vitamin C in the diet, the combination may prove to be more beneficial than either alone. The combination of citrus extract and vitamin C was found to produce a synergistic antioxidant effect in an in vitro lipoprotein oxidation model. In a double-blind, placebo-controlled study with 26 normal and hypercholesterolemic subjects, the citrus extract and vitamin C, but not vitamin C or vitamin E alone, significantly lowered triglycerides. The combination of citrus extract and vitamin C increased the lag time of lipoprotein oxidation, compared with vitamin C alone or a placebo, and was a significantly better antioxidant than vitamin E. These results and other published studies are highly suggestive of in vitro and in vivo antioxidant synergism between citrus extract and vitamin C. PMID- 12639399 TI - Protective effect of soybean saponins and major antioxidants against aflatoxin B1 induced mutagenicity and DNA-adduct formation. AB - Saponins from various plant sources have been suggested as possible anticarcinogens. Major dietary sources of saponins include legumes such as soybeans. This study was performed to determine the effect of soybean saponins on aflatoxin B(1)(AFB(1))-induced mutagenicity and AFB(1)-DNA adduct formation using Salmonella typhimurium and human liver hepatoma (HepG2) cells, respectively. Major antioxidants including L-ascorbic acid, alpha-tocopherol, all-trans retinol, and butylated hydroxytoluene (BHT), previously reported to possess antimutagenic activity, were used as test materials to evaluate the relative effectiveness of saponins. Results indicated antimutagenicity was in the order of BHT > saponins > alpha-tocopherol > L-ascorbic acid. Soybean saponins exerted a significant effect, inhibiting the mutagenicity of AFB(1) by 52%, 64%, and 81% at concentrations of 600, 900, and 1,200 microg per plate, respectively. The amount of tritiated AFB(1) metabolites-DNA adducts formed in HepG2 cells was significantly reduced when cells were preincubated with 10 or 30 microg/ml of test materials. Soybean saponins inhibited AFB(1)-DNA adduct formation by 50.1% at a concentration of 30 microg/ml, whereas L-ascorbic acid and BHT reduced adduct formation by 38.4% and 32.6%, respectively, at the same concentrations. These results indicate that soybean saponins possess not only a significant antimutagenic activity but a strong inhibitory action against carcinogen-induced DNA damages. Soybean saponins possibly block the initiation stage of carcinogenesis, and further studies are required to elucidate the mechanisms of action. PMID- 12639401 TI - Role of Spirulina in the Control of Glycemia and Lipidemia in Type 2 Diabetes Mellitus. AB - Spirulina, with its high concentration of functional nutrients, is emerging as an important therapeutic food. This study aimed to evaluate the hypoglycemic and hypolipidemic role of Spirulina. Twenty-five subjects with type 2 diabetes mellitus were randomly assigned to receive Spirulina (study group) or to form the control group. At baseline, the control and study groups were matched for various variables. The efficacy of Spirulina supplementation (2 g/day for 2 months) was determined using the preintervention and postintervention blood glucose levels, glycosylated hemoglobin (HbA(1c)) levels, and lipid profiles of the diabetic subjects. Two-month supplementation with Spirulina resulted in an appreciable lowering of fasting blood glucose and postprandial blood glucose levels. A significant reduction in the HbA(1c) level was also observed, indicating improved long-term glucose regulation. With regard to lipids, triglyceride levels were significantly lowered. Total cholesterol (TC) and its fraction, low-density lipoprotein cholesterol (LDL-C), exhibited a fall coupled with a marginal increase in the level of high-density lipoprotein cholesterol (HDL-C). As a result, a significant reduction in the atherogenic indices, TC:HDL-C and LDL-C: HDL-C, was observed. The level of apolipoprotein B registered a significant fall together with a significant increment in the level of apolipoprotein A1. Therefore, a significant and favorable increase in the ratio of A1:B was also noted. These findings suggest the beneficial effect of Spirulina supplementation in controlling blood glucose levels and in improving the lipid profile of subjects with type 2 diabetes mellitus. PMID- 12639402 TI - Pycnogenol((R)) in the Management of Asthma. AB - Asthma is characterized as a chronic inflammatory process. Pycnogenol((R)), a bioflavonoid mixture extracted from Pinus maritima, is known to scavenge free radicals while possessing antioxidant and antiinflammatory properties. The objective of this study was to evaluate the efficiency of this agent in a randomized, double-blinded, placebo-controlled, crossover study in patients with varying asthma severity. Twenty-six patients who fulfilled the American Thoracic Society criteria for asthma were enrolled in the study. Medical history, physical examination, blood sample analyses, and spirometric values were obtained at baseline, 4 weeks, and 8 weeks. The patients were randomly assigned to receive either 1 mg/lb/day (maximum 200 mg/day) Pycnogenol or placebo for the first period of 4 weeks and then crossed over to the alternate regimen for the next 4 weeks. No adverse effects were observed related to the study drug. Within the contingent of 22 patients who completed the study, almost all responded favorably to Pycnogenol in contrast to placebo. Pycnogenol treatment also significantly reduced serum leukotrienes compared with placebo. The results of this pilot study indicate that Pycnogenol may be a valuable nutraceutical in the management of chronic asthma. We recommend that further clinical trials be conducted in larger groups of asthmatics to establish its efficacy. PMID- 12639403 TI - Antioxidant Activity of Anthocyanin Extract from Purple Black Rice. AB - Antioxidant activity was studied for anthocyanins extracted from purple black rice (PBR) by a 3% aqueous trifluoroacetic acid solution (TFA), as well as for anthocyanins extracted from blueberry (Bluetta, high bush type). Capillary zone electrophoresis revealed that the PBR extract contained almost exclusively a single anthocyanin, which was identified as cyanidin 3-O-beta-D-glucoside (Cy 3 Glc) after purification by polyvinylpyrrolidone column chromatography. In contrast, 11 anthocyanins were identified in the blueberry extract. PBR extract showed slightly weaker superoxide scavenging and crocin bleaching activities than blueberry extract did. Both PBR and blueberry extracts, however, showed 10 to 25 times stronger activity than the same concentration of Trolox used as a reference antioxidant. It was further noted that the purified Cy 3-Glc from PBR extract retained approximately 74% of the antioxidant activity (both crocin bleaching and superoxide scavenging) observed in the original TFA extract. The hydroxyl radical scavenging activity of both extracts was several times weaker than that of the same concentration of Trolox, although the PBR extract showed approximately two times stronger activity than blueberry extract did. The hydroxyl radical scavenging activity of the purified Cy 3-Glc from PBR, however, decreased to approximately 20% of that of the original PBR extract. These results indicate that the anthocyanin Cy 3-Glc contributes to the antioxidant activity of PBR through its strong superoxide radical but not hydroxyl radical scavenging activity. PMID- 12639404 TI - Chain-Breaking Antioxidant Capability of Some Beverages as Determined by the Clark Electrode Technique. AB - The protective effects of red wine, tea, and coffee on cancer, atheroclerosis, and other diseases are attributed to the antioxidant activity (AOA) of polyphenols, which are abundant in those beverages. We present a rational procedure for determining the total chain-breaking AOA of beverages with the use of the Clark electrode technique. The procedure is based on the steady monitoring of oxygen consumption accompanying the chain peroxidation of methyl linoleate in Triton X-100 aqueous micelles induced by 2,2'-azobis(2-amidinopropan) dihydrochloride as a source of active free radicals. AOA was characterized by the number of kinetic chains (expressed in concentration units) that could be terminated by a beverage. The procedure was applied to determine the AOA of nine red wines and single samples of green and black teas, white wine, beer, and soluble coffee. The addition of any of the studied beverages to the testing system resulted in pronounced retardation of methyl linoleate oxidation. The period of increase in the oxidation rate with time caused by antioxidant consumption was preceded by a period when the oxidation rate was visibly decreased with time (increase in inhibition). The release of polyphenols from their complexes with other components of beverages was suggested as the most probable mechanism of this event, which was observed for the first time in this study. PMID- 12639405 TI - Glucose but Not Polypeptone Reduces the Effect of Probiotic Preparations to Stimulate Carbohydrate Breakdown and Reduce Net Ammonia Production by Pig Cecal Bacteria In Vitro. AB - Probiotic preparations are used to prevent or treat diarrhea. Probiotic preparations increase the in vitro breakdown of carbohydrates and decrease that of protein by mixed cecal bacteria in the absence of readily fermentable materials. Diarrhea can increase the influx of readily digestible materials into the large intestine. Therefore, we compared production of organic acids and ammonia in batch cultures using pig cecal contents with or without probiotic preparations (Clostridium butyricum, Lactobacillus casei plus Bifidobacterium breve, or L. casei plus Enterococcus faecalis) and with or without readily fermentable substrates (glucose or polypeptone). Effects of additional substrates appeared earlier than those of probiotic preparations. Both probiotic preparations and glucose increased the production of most short-chain fatty acids and lactic acid but decreased those of ammonia and isovaleric acid. Effects of probiotic preparations and glucose were not additive. Effects of different preparations were alike. Polypeptone increased the production of ammonia and isovaleric acid independent of probiotic preparations. Effects of probiotics and glucose may be to provide either energy or carbon to the microbial ecosystem. Effects of glucose were probably mediated through their direct utilization by indigenous bacteria. PMID- 12639406 TI - Immunomodulatory Properties of Cucumariosides from the Edible Far-Eastern Holothurian Cucumaria japonica. AB - Holothurian triterpene glycosides (cucumariosides) are known to possess multiple biological activities. Here we show that cucumariosides from the Far Eastern edible holothurian (sea cucumber), Cucumaria japonica, and their semisynthetic derivatives possess potent immunomodulatory properties. Intraperitoneal injection of cucumariosides (0.2-20 ng per mouse) induced macrophage lysosomal activity in a dose-dependent manner (up to 250% of control). The stimulatory effect was related to the chemical structure of cucumariosides and was especially influenced by the number and position of sulfate groups in the carbohydrate moiety of the molecules. In vitro, an inhibitory rather than a stimulatory effect of cucumariosides on phagocytosis and release of tumor necrosis factor-alpha (TNF alpha) was found. Virtually all cucumariosides inhibited latex bead phagocytosis by human peripheral blood granulocytes in a dose-dependent manner. Also, the lipopolysaccharide-induced TNF-alpha production by immune cells in human blood diluted with RPMI-1640 was decreased, without a clear relation to the structure and dose of the compounds. The data are discussed in terms of possible mechanisms underlying immunomodulatory properties of triterpene glycosides from C. japonica. PMID- 12639407 TI - Correct Identification of Passiflora incarnata Linn., a Promising Herbal Anxiolytic and Sedative. AB - Passiflora incarnata Linn. and Passiflora edulis Sims are the two important plants of the family Passifloraceae that have often been reported as synonymous because of their identical morphological and microscopic characteristics. P. incarnata is a popular sedative and anxiolytic, whereas, P. edulis is rarely reported to possess significant central nervous system depressant activity. P. edulis, as the name of the species reflects, is mainly grown for edible purposes. During a survey of literature on the genus Passiflora, it was noticed that in many references the two plants are mentioned synonymously. The designation by Sir William J. Hooker in 1843, followed by the citation of P. edulis as the synonym of P. incarnata in Index Kewensis of 1895, not only substantiated the controversial identity but also caused confusion to researchers. The prevailing confusion might have led to improper selection of the bioactive plant, thereby accounting for inconclusive and contradictory pharmacological reports on either of the two plants. In this work, we establish key identification parameters to differentiate P. incarnata from P. edulis. Various leaf constants such as vein islet number, vein-termination number, stomatal number, and stomatal index are different for the two species. Physicochemical parameters such as ash values and extractive values and the thin layer chromatography profile of the petroleum ether extract of P. incarnata and P. edulis are also distinct and different. Various clinical uses of P. incarnata for anxiety and allied diseases are discussed. PMID- 12639408 TI - Relation of Soy Consumption to Nutritional Knowledge. AB - Is it reasonable to believe that the consumption of medicinal and functional foods such as soy is driven by nutritional knowledge? A national survey of 770 U.S. consumers indicated that 39% of male and female subjects did not know of any health benefits associated with soy. Among those who know of soy's nutritional benefits, consumption occurred only among those who perceived these benefits as specifically relevant to themselves. People who had even a slight familiarity with functional foods were more likely to consume soy. Improving the taste properties of soy appeared to have a more dramatic potential impact on the consumption of those who were knowledgeable about functional and medicinal foods than on the general population. PMID- 12639409 TI - Clinical Effects of a Dietary Antioxidant Silicate Supplement, Microhydrin((R)), on Cardiovascular Responses to Exercise. AB - Amorphous silicate minerals, often described as rock flour, were once common in natural water sources and abundant in glacial stream waters. Not only do the silica mineral particles bond water and other elements for transport; they also can be adsorbed with reduced hydrogen, which releases electrons, providing antioxidant or reducing potential to surrounding fluids. The purpose of this investigation was to examine the cardiovascular responses during exercise after consumption of a dietary silicate mineral antioxidant supplement, Microhydrin((R)) (Royal BodyCare, Inc., Irving, TX). A clinical trial incorporating a double-blind, placebo-controlled, crossover experimental design was employed. Subjects received either active agent or placebo, four capsules per day, for 7 days before the trial. The trial evaluated six exercise bicycle trained subjects performing a 40-km bicycling time trial. Ratings of perceived exertion and measurements of oxygen uptake, heart rate, performance workload, and preexercise and postexercise blood lactate concentrations were obtained. Although there were no differences (P >/=.05) in work performed, heart rate, oxygen uptake, and ratings of perceived exertion during the time trial, the postexercise blood lactate concentrations were significantly lower (P GSE2 > GSE1 > dimethylsulfoxide. The higher antioxidant capacity and anticlastogenic activity of GSEs can be explained, structurally, by the high number of conjugated structures between the catechol groups in the B rings and the 3-OH free groups of the polymeric polyphenolic skeleton and, in addition, by the stability of the aroxyl flavonoid radical generated in the processes. PMID- 12639421 TI - [General introduction to Examining Committee for Physician of traditional Chinese medicine (TCM) in the 1940s]. AB - Established in 1944, the Examining Committee for Physician was the outcome of struggle of TCM physicians seeking for survival in the Republic of China. The committee comprised persons from the selecting committee of government and the TCM committee of Health Office. Its functions were examining the qualification of practitioners of TCM who applied, according to the qualified conditions of TCM physician decided by Examine Office, and sent out the certification to qualified TCM physicians for the eligibility of practising; laid down and mastered detailed rules of the eligible conditions of TCM physician; laid down the principles and applied interview; participated in the operation of TCM examination and made relevant decisions and suggestions; measured and managed concrete problems in examining process, etc. PMID- 12639422 TI - [Strengthening process of the licensing system for physicians in the Republic of China]. AB - The Provisional Regulation for Administration of Physician, issued by the Internal Affairs Department of the Northern Warlords Government on March 9, 1922; The Provisional Regulation of Physician, issued and carried out by Ministry of Health of Nanjing National Government on January 15, 1929; and The Provisional Regulation of Physician, issued and carried out by Executive Yuan of Nanjing National Government on August 8, 1940 marked preliminary formation of the licensing system for physicians. The Standards of Physician, issued and carried out by Nanjing National Government on September 22, 1943 marked formation of the licensing system for physicians in the Republic of China. In the later stage of the War of Resistance Against Japan, some legal documents, like Detailed Rules of the Standards of Physician etc. issued and carried out by Nanjing National Government were the additional remarks for the administration of the Standards of Physician. All of these showed that the licensing system for physicians was strengthened step by step. PMID- 12639423 TI - [A study of the licensing system in Korean Oriental medicine]. AB - In 1952, the Licensing System of Korean Oriental Practitioners was established by law and practiced until now. It was the most important aspect of the Professionalization of Korean Oriental medicine, and it also affected other aspects of the Professionalization of Korean Oriental medicine, such as the standardization of knowledge, the strengthening of its organization, and the development of an occupational ideology. The Licensing System of Korean Oriental Practitioners came about as a result of conflicts between the National Assembly, governmental agencies, and medical doctors. The range of Korean Oriental Practitioners Licensing is partially limited, especially with respect to certifying someone as a public health doctor and in conferring the right to direct medical technicians. However, the limitations have been reduced gradually. PMID- 12639424 TI - [A study on Cai Xishan's Mai jing (Pulse classic)]. AB - Cai Yuanding, also named Cai Xishan, was a great Confucian scholar in the Southern Song dynasty, and his book, Mai jing (Pulse Classic), was long lost in China. According to Dai Qizong's Mai jue kan wu ji jie (Collected Annotations of Correcting Errors in Pulse Study in Verse) of the Yuan dynasty, Liu Yude's Yi lin xu chuan (Extending Biography of Medicine) of the Ming dynasty etc., the author makes a study on "Pulse Classic", the fourth volume of Zhen mai xu zhi (Notice of Pulse Taking), a book came back from the Cabinet Library of Japan and holds that it was an extract edition of Cai's Pulse Classic. When discussing the pulse, Cai paid attention to the theory of pulse while neglected the pulse conditions and its indications, which was further researched by the author. PMID- 12639425 TI - [Discussion on Tang edition (TE) and notes of the Tang edition (NT) under the "ink cap" in Zheng lei ben cao (Classified materia medica)]. AB - The TE and NT cited under ink cap in Zheng lei ben cao (Classified Materia Medica) cannot be found in the contents and fragmentary volumes of Xin xiu ben cao (Newly Revised Materia Medica), yet can be seen in the "annotations of Shu ben cao (Materia Medica of Sichuan)", cited by Zhang Yuxi. This showed that the TE and NT were, in fact, coming from Shu ben cao, whose old name was Chong guang ying gong ben cao (Augmented Yinggong's Materia Medica). The so - called Yinggong was Li Ji, who compiled Tang ben cao (Materia Medica of the Tang Dynasty) by the imperial order, and gained the title Master Yingguogong, hence the title. Chong guang ying gong ben cao was meant revised Tang ben cao. This was the reason why Tang Shenwei called Shu ben cao as Tang ben cao right away in his Zheng lei ben cao. PMID- 12639426 TI - [Collation and research on the different texts in Qian jin yao fang (Precious essential formulary, PEF)]. AB - There is considerable difference in the texts between PEF and the archaic editions Sun zhen ren qian jin fang (The Immortal Sun's Precious Formulary, ISPF) and the Song - revised edition Bei ji qian jin yao fang (Precious Essential Formulary for Emergency, PEFE). It is hoped that the real features of PEF can be worked out through collated studies on these two editions so as to offer a basis for systematic research on PEF. At the same time, it has been proved that during the revision and collation of PEF by the Song scholars, it is certain that dramatic revisions had been made on its medical materials, especially an abundant amount of medical materials were added. As a result, the current circulated edition of PEFE does have obvious differences with Sun's original work, while ISPF, in contrast, preserves the original features of Sun Simiao. Hence, ISPF is of important academic significance in the in - depth study of PEF. PMID- 12639427 TI - [Science of acupuncture and moxibustion in Si bu yi dian (rGyud - bzhi)]. AB - There are plentiful contents in the science of acupuncture and moxibustion in Si bu yi dian (rGyud - bzhi), which absorbs much knowledge of acu - moxibustion from Central Plains, and forms its own system. There are unique and perfect theories, and abundant techniques in its contents. Great attention was paid to therapeutic method of pyro - moxibustion, and some amazing and mature achievements are created, such as eliminating cataract with bronze needle etc. Especially, there are rather high attainments in the research of blood - letting. rGyud - bzhi is a great book of minority medicine, and helpful for the further study and development of modern science of acu - moxibustion. PMID- 12639428 TI - [Origin and influence of academic contending from cold school and warm school of paediatric]. AB - The academic contending from cold school and warm school of pediatrics was derived from Qian Yi's and Chen Wenzhong's therapeutic opinions for measles and smallpox in the Song dynasty. The former claimed the use of cold and cool drugs, while the latter, maintained the use of warm and hot drugs. During the Jin, Yuan, Ming and Qing dynasties, physicians continually revised and supplied their therapeutic methods and started the academic contending in almost all fields of pediatrics, including therapeutic opinion for measles and smallpox, exogenous pathogenic diseases of children, miscellaneous diseases of children etc., which improved the further research on basic theory of pediatrics, such as constitution, physiological and pathological characteristics of children, and also pushed forward the academic development of pediatrics of traditional Chinese medicine. PMID- 12639429 TI - [The importation of the first X-ray diagnostic machine in China]. AB - There are different opinions and controversies about the first importation of X - ray diagnostic machine, which was imported into China. Dian shi zhai hua bao (Pictorial of Dianshizhai), based on new historical materials, reported that Boxi Hospital of Suzhou imported the X - ray diagnosed machine before the last ten day of December, 1897. PMID- 12639430 TI - [A brief history of hospital development in the West]. AB - Origin and development of hospital in the West was related closely with religions and wars. Before the Medieval Ages, priests or physicians treated their patients mainly in temples, which may be regarded as the earliest type of hospital. The military hospital first appeared in the ancient Rome due to war. During the Medieval Ages, most hospitals were attached to monasteries. The Crusades prompted the development of hospitals and hospices along the route taken by the Crusaders. After the Medieval Ages, as the Protestantism rose, the monastic hospitals suffered a major setback and secular hospitals developed. From the 16th century, hospitals of Europe were spread gradually into North America. During the 18th and 19th centuries, function of hospital in medical treatment, education and research was strengthened gradually. In the 20th century, hospital facilities were matured technically and management of hospital became an important problem to which people devoted themselves to resolve it. PMID- 12639431 TI - [Investigation on folk medicine and health care of She ethnic group in Zhejiang province]. AB - Investigation has been made on the folk medicine and health care in Jingning Autonomous Prefecture of She ethnic group, the only of its kind in China. It is demonstrated that since the living condition is bad, the hygienic environment is very poor, epidemics like measle, malaria and venereal diseases are very prevalent, and a high incident area of hyperthyroidism. Since there are no special writing characters, She ethnic group spread their medical know - how by oral transmission. Collection of herbs and compounding of recipes are mainly through the family. Medicine of the She ethnic group has a long history, and the treatment of bone - injury, snake - bites and wind - pain are very unique. It has been found that there are 599 genuses, 199 families of herbs in She medicine with 1043 species; among them, 214 herbs are in common use, falling under 126 genuses. In addition, 7 new herbal plants are found, and 332 recipes collected. Currently, over 40 She physicians are practicing in the prefecture. PMID- 12639432 TI - [Textual research and explanation of "Qi - Huang"]. AB - Since the main contents of the Huang di nei jing (Yellow Emperor's Inner Canon) was completed in the Warring States, in the form of questions and answers between Huang Di and Qi Bo, Qi - Huang is used as the appellation for Yellow Emperor's Inner Canon in later generation. As the Yellow Emperor's Inner Canon laid the foundation for the development of traditional Chinese medicine (TCM), in the Song Dynasty, Qi Bo and Huang Di was jointly called Qi - Huang for the first time in Bian que xin shu (Medical Understanding of Bian Que) by Dou Cai. And they were also worshipped as the orthodox ancestors of medicine in the shrine with new implication. Since then, Qi - Huang had an extended meanings for orthodox TCM or TCM science. However, in most cases, Qi - Huang was the appellation of TCM and TCM science. Meanwhile, new words derived from Qi - Huang have their respective meanings, such as "the art of Qi - Huang" and "the theoretical principles of Qi - Huang" standing for the science of TCM or the art of healing, and the theoretical principles. As for the "Qi - Huang physicians", it represents the TCM doctors and specialists. Dan xi shou jing (Medical Mirror of Danxi) by Wu Shangmo in the Ming Dynasty says that "meeting Qi - Huang specialists". "Qi - Huang books" means TCM books. And Qi - Huang profession refers to TCM profession, etc. It can be seen that the term Qi - Huang does have a rich content of Chinese traditional culture. PMID- 12639433 TI - [Study on Shi Zhenduo's Ben cao bu (Supplemented materia medica)]. AB - Being called "precursor of pharmacy imported from the west" by Fan Xingzhun, a famous medical historian, Ben cao bu was lost in China. It has been found outside China recently. This article deals with the author, blockprinting edition, contents of the book and makes a comparison with its texts cited in Ben cao gang mu shi yi (Supplemented Compendium of Materia Medica). It also evaluates the practical significance of this book as an early dissemination of pharmacy imported from the west. PMID- 12639434 TI - [Circulation of Song editions of Hong shi ji yan fang (Hong's Collection of effective recipes) and Shang han yao zhi yao fang (Recipes with gist on cold pathogenic diseases)]. AB - The first blockprinted edition, 5 - volume Hong shi ji yan fang (Hong's Collection of Effective Recipes) in the 6th year of Qiandao reign (1170) and the first blockprinted edition of Shang han yao zhi yao fang (Recipes with Gist on Cold Pathogenic Diseases) in the 7th year of Qiandao reign (1171) are collected in the National Beijing Library. Both are highly treasured by book connoisseurs of the Ming and Qing dynasties who made much textual research. The authors, edition uniqueness and circulating facts of both works are discussed here. PMID- 12639435 TI - [Research on original editions of Lei jing (Classified canon)]. AB - After comparing with different extant editions of Nei jing (Inner Canon), the author holds that the portion of Su wen (Plain Questions) in Lei jing (Classified Canon) was similar to Hu's 12 - volume system of Gulinshutang edition, Zhao's Jujingtang edition, while its portion of Ling shu (Miraculous Pivot) is closer to the proofed edition of Zhou's and Ma Shi's annotated 9 - volume edition. After his determination of the original edition, he further made collation with comparison with other editions, and checked the original texts accordingly. Hence it was different from all other available editions. PMID- 12639436 TI - [A preliminary analysis on the detoxified recipe and medicines for aconite intoxication in Wu shi er bing fang (Formularies for 52 kinds of disorders)]. AB - As an excellent analgesic, aconite is highly toxic, hence, its detoxification has become an important topic to the medical professionals, ancient and modern. There is a rich contents with detailed descriptions, and many medicines for the detoxication of aconite in Wu shi er bing fang (Formularies for 52 Kinds of Disorders), being very unique in its method. It not only gives valuable experiences, but also opens an earliest example to detoxify aconite intoxication for later agas. PMID- 12639437 TI - [On the connotation of "Shennong tastes all kinds of herbs and encounters seventy kinds of toxic substances in a single day"]. AB - The description of "Shennong tastes all kinds of herbs and encounters seventy kinds of toxic substances in a single day" is a legend in Chinese medical history, demonstrating that medicine is originated from practice. This description has been interpreted as a semi - mythological legend. In fact, the description has a true and primitive background. Among them, the "day" and "seventy" bear specific implication and deep historical and cultural connotation. As time progresses, the fading of history will be even more deified. Hence, this phenomenon should arouse attention during the study of medical history. PMID- 12639438 TI - [Huang Zongxi and medicine]. AB - Being a very famous Confucianist, Huang Zongxi also had indepth understanding in medicine. When dealing with the famous physicians of his native land in eastern Zhejiang province, such as Zhang Jiebin, Zhao Xianke, Gao Doukui, Huang, in addition to his positive appraisal on these persons' contribution to the theory of Warm diseases, particularly Zhang Jiebin, also made comments dialectically on their detrimental effect of the Theory of Warm - benefiting on the society and the rash action of denying the theory of cold pathogenic disorders and even ignoring channel - collaterals, but biasing to only one channel, particularly that carried in Zhao Xianke's Yi Guan (Medical Permeation). PMID- 12639439 TI - [The early magazines of traditional Chinese medicine in Fuzhou region]. AB - The early magazines of traditional Chinese medicine (TCM) in Fuzhou region, Xian dai yi yao (Modern Medicine) and Yi duo (Medical Bell) did play certain historical role in the protection of TCM, dissemination of Chinese medical know - how, and promotion of academic exchanges. PMID- 12639440 TI - [A brief history of evidence-based medicine]. AB - Evidence - based medicine, a fairly new subject, focusing on the perfect combination of individual clinical experience, existing best evidence available and patient value, has created a new approach and thought for clinicians and researchers. Evidence - based medicine has promoted the transition of health care. The definition and development of evidence - based medicine in China and the world as a whole are reviewed in this article. PMID- 12639441 TI - [Developmental history of the cruciate ligament surgery]. AB - The study of the cruciate ligaments have been made since 3000 B. C., including anatomical observation, functional description, examination and treatment approaches. The developmental history of the cruciate ligaments is the advances of human ideas and techniques in the area generations after generations. PMID- 12639442 TI - [Influence of Neo-Confucianism in the Song and Ming dynasties on the theory of traditional Chinese medicine]. AB - The academic style of study and thoughts of Neo - Confucianism in the Song and Ming dynasties improved the innovation of contemporary physicians and their recognition on the research of medical theories, and strengthened the development of determination of treatment based on the differentiation of symptoms and signs. Its related contents, such as taiji, circulation and change of vital energy, substance and functions, and congenital and postnatal etc. also were absorbed as the elements of medical theories. However, there were some negative influences from Neo - Confucianism on the development of traditional Chinese medicine. PMID- 12639443 TI - [Two characteristics on the developmental history of art of prescription]. AB - By recalling the history of art of prescription and summarizing its law of development, two apparent characteristics can be shown on its primitive accumulation and clinical treating thoughts. One is the trend of transformation from empirical formula to theoretical formula, and from the discrepancy between medical theories and medical prescriptions to their combination. The other one is from treatment based on differentiation of diseases to differentiation of symptoms and signs, and from correspondence of prescription and diseases to correspondence of prescription and syndrome. It is of realistic significance to further recognize the developmental characteristics of the art of prescription, not only for mastering the prescription rules and main points of matching of ingredients in compounding, but also on comprehending its scientific implication and mechanism. PMID- 12639444 TI - [Textual research on Shi Kan and Shi zai zhi fang (Shi Zaizhi's Prescriptions)]. AB - Shi Kan, also known as Zaizhi, was born in Emei, Sichuan, and Zaizhi was more popularly known. He lived in the Northern Song dynasty, and was a successful candidate in the highest imperial examinations. As a government official, he was promoted to the position of governor. When he cured the constipation of Cai Jing only with a single herb Aster root, people considered that he had excellent medical skill as famous doctor Xu Shuwei. He was one of the representatives who were doctors as well as government officials. His book Shi zai zhi fang (Shi Zaizhi's Prescriptions) was a typical formula work of the Song dynasty. In this book he emphasized the treatment principles on strengthening the vital and dispelling the evil. He also underlined the theory of yunqi (five elements' motion and six kinds of natural factors). He had many new ideas about the application of herbs and pulse - taking. According to the book, Zhi zhai shu lu jie ti (Zhizhai's Interpretation on Collected Books), Shi had a 2 - volume book titled Zhi nan fang (Guide Prescriptions). There were different views on whether Guide Prescriptions was the other name for Shi Zaizhi's Prescriptions. By studying the contents of citations from Shi's Guide Prescriptions quoted in Hong shi ji yan fang (Hong's Collections of Proved Recipes), Quan sheng zhi mi fang (A Guidebook of Recipes for Saving Lives), and by combining it with the textual research made by various scholars of the Qing dynasty, the author claims that Guide Prescriptions is another book rather than the other name of Shi Zaizhi's Prescriptions. PMID- 12639445 TI - [Xu Ren's Zhen jiu jing yan fang (Experiential recipe on acupuncture and moxibustion) and its spread]. AB - The book, Zhen jiu jing yan fang (Experiential Recipe on Acupuncture and Moxibustion), written by Xu Ren, an imperial physician of Korea was completed in the 4th year of Renzu emperor (1644), and was an important work for the history of Korean acupuncture and moxibustion. However, there were mistakes on its writing date as described in the books of acu - moxibustion history in former reports. The portion of Mian xue tang zhen jiu ji cheng (Mianxuetang's Complete Work of Acupuncture and Moxibustion) universally recognized to be unique, was plagiarized from Zhen jiu jing yan fang. PMID- 12639446 TI - [Discussion on academic origin of Ao shi shang han jin jing lu (Ao's Records of golden mirror on cold pathogenic diseases]. AB - Ao shi shang han jin jing lu (Ao's Records of Golden Mirror on Cold Pathogenic Diseases) is the extant first work of lingual diagnosis. It is demonstrated that the author of this book is categorized under the school of Liu Wansu based on the location of disease, the nature of disease, therapies and drugs, and pathogenesis in pathological lingual pictures described in this book. The lingual diagnosis was based on Liu Wansu's theory of fire - heat, and used for diagnosing warm diseases. Intuitionistic vision was used in this book to prove the direct relation between red tongue and heat syndrome, which was the important joint for the theory of warm diseases to derive from the theory of cold pathogenic diseases. At the same time, the set up of the model of diagnosis and treatment for red tongue - inner heat syndrome, and clearing inner heat is established. PMID- 12639447 TI - [Study on Wang Ruzun's hand - written copy of Dongyi Bogam (Precious mirror of oriental medicine)]. AB - In the beginning of the 16th century, Hu Jung of the Lee dynasty of Korea collected the chief medical books written before the mid - Ming dynasty of China and some ancient Korean medical books to compile the Dongyi Bogam (Precious Mirror of Oriental Medicine). This marks the formation of a new stage in Korean medicine with unique theoretical system. After this great book was transmitted to China, the first man who tried to blockprint it was a folk doctor, Wang Ruzun. This article briefly introduces Wang's life, gives a general picture of Wang's hand - written copy in 1747, by quoting Chai Chaosheng's preface and Wang Ruzun's postscript in the book. It is firmly believed that Wang's hand - written copy is of great significance. The said copy is collected in our library. PMID- 12639448 TI - [Major surgical achievements in the period from the Warring States to the Three Kingdoms]. AB - The period from the Warring States to the Three Kingdoms was the stage of laying foundation preliminarily for surgical theory of traditional Chinese medicine (TCM). Observing and analyzing it from two aspects, that is development of the clinical practice of surgery and the establishment of the basic theory of surgery, it is helpful for recognizing its process of establishment, development and improvement. PMID- 12639449 TI - [Historic significance of Hua Tuo's theory of "Six parts" and "Three methods" in cold pathogenic diseases]. AB - In the end of the Eastern Han dynasty, Hua Tuo and Zhang Zhongjing explored deeply and separately the diagnostic and therapeutic principles of cold pathogenic diseases, though they didn't know each other. This was not only because of their excelsior spirit, but also showed the severity of cold pathogenic diseases at that time. The doctors in past dynasties researched and inherited the achievements of Zhongjing's Shang han lun (Treatise on Cold Pathogenic Diseases). The theory of Hua Tuo's "transformation of diseases by six parts" was different from Su wen re lun (Discussion on Heat in Plain Questions). It doesn't put 6 - channel theory equally, but giving a series of rules for diagnosing the syndromes transforming from superficies to interior, from the surface to the bottom, and from upper to lower which is the beginning of wei - qi - ying - xue differential diagnosis and three - jiao differential diagnosis. Hua Tuo treated cold pathogenic disorders by "sweating, emesis, and catharsis" with abundant clinical approaches. Hua Tuo's idea of "rotten stomach with petechiae" was valued by scholars of warm disorders and of clinical significance even today. PMID- 12639450 TI - [Centenary history of College of Medicine, National Taiwan University]. AB - Orthodox medical education in Taiwan dates back to the beginning of the Japanese occupation. The Japanese first established a hospital in Taipei (Taihoku in Japanese), and created a "Medical Teaching Institute for Taiwanese", which was the embryonic form of medical education in the period of Japanese occupation. In 1899, a formal institution of medical education was established in the form of the Medical School of the Governor - Generalship of Taiwan. Later, in 1919, it was renamed the Medical College of the Governor - Generalship of Taiwan, and in 1922, its name was again changed to Taihoku Medical College. In 1936, Taihoku Imperial University created a school of medicine, which in 1945 after the liberation from the Japanese was renamed the College of Medicine, National Taiwan University, whose first dean was Dr. Tu Tsung - Ming. At the same time, Taihoku Imperial University Hospital was renamed National Taiwan University Hospital. The College of Medicine, National Taiwan University was the only institution of medical education after the island was restored to Chinese control. During the past 50 years, medical education has undergone major change and development. By 1990, the number of medical colleges in Taiwan had increased to eleven. This paper described the College of Medicine, National Taiwan University and its hospital before and after the restoration, the background and development of medical education in Taiwan. It also describes the College's achievements in the realm of education, research and healthcare trends. PMID- 12639451 TI - [A brief account on the characteristics of confluence of Chinese and Western medicine in medical affairs of modern Shanxi]. AB - Shanxi is one of the most active provinces in China for its research on modern traditional Chinese medicine (TCM), especially on medical practice of confluence of Chinese and Western medicine, where many distinguishing features exist and are deserved to study. This paper gave a brief account on the characteristics of confluence of Chinese and Western medicine in medical affairs of modern Shanxi from the following 5 aspects: the policy and situation of confluence of Chinese and Western medicine; the Promoting Research Committee of Shanxi TCM for promoting TCM; Shanxi Teaching Institute of Medicine where some courses about confluence of Chinese and Western medicine are taught; Medical Magazine for the confluence of Chinese and Western medicine; famous medical workers of confluence of Chinese and Western medicine in Shanxi. PMID- 12639452 TI - [Observation on and naming to plague in folk people and medical practitioners of modern China]. AB - It is helpful to differentiate plague from other infectious diseases from the point of view of ordinary people and medical practitioners, by observing and studying how they observed the activities of rats, described symptoms of patients, and named plague. Although they had well known about such knowledge, they did not distinguish this disease from others and various names were existed. PMID- 12639453 TI - [Historical review on treatment of head and neck cancer in modern medicine]. AB - The summarization includes understandings of the etiological factors, evolution course of treatment of head and neck cancer in pre - modern era (1500 - 1900) and modern age, and the historical summary of head and neck cancer treatment in China, aiming at providing references for its further development through historical reviewing on its law summarized through 150 years of experience. PMID- 12639454 TI - Atopic dermatitis. AB - Atopic dermatitis, the dermatologic manifestation of the atopic diathesis, has a variety of clinical presentations. This disease probably should be considered a syndrome--a group of signs and symptoms that frequently occur together in an identifiable pattern. The following update describes the spectrum of atopic dermatitis and summarizes current thinking about the etiology of the disease. PMID- 12639455 TI - Molluscum contagiosum. AB - Molluscum contagiosum is a disease caused by a poxvirus of the Molluscipox virus genus that produces a benign self-limited papular eruption of multiple umbilicated cutaneous tumors. This common viral disease is confined to the skin and mucous membranes. Transmission requires direct contact with infected hosts or contaminated fomites. It is generally thought to infect humans exclusively, but there are a few isolated reports of Molluscum contagiosum occurring in chickens, sparrows, pigeons, chimpanzees, kangaroos, a dog, and a horse. The infection is found worldwide and has a higher incidence in children, sexually active adults, and those who are immunodeficient. PMID- 12639456 TI - Pimozide: use in dermatology. AB - Pimozide is widely used in psychiatry for chronic psychoses, schizophrenia, the syndrome of Gilles de la Tourette and to a certain extent, also in dermatology. The only dermatological indication is for delusions of parasitosis. Though there is a good rationale for using pimozide in this disease, the majority of the studies on pimozide in dermatology are uncontrolled trials and case reports. PMID- 12639457 TI - Cimetidine: a review of the recent developments and reports in cutaneous medicine. AB - Cimetidine, approved by the FDA for inhibition of gastric acid secretion, has been advocated for a number of dermatological diseases. The cutaneous uses and immunological effects of cimetidine have been actively studied over the past few years, and this review summarizes the literature accumulated since 1997. PMID- 12639458 TI - Recent advances in the treatment of leprosy. AB - Leprosy, a chronic infectious disease caused by Mycobacterium leprae, was identified by G. H. A. Hansen in 1873. The different clinical presentations of the disease are determined by the quality of the host immune response. The bacteria have affinity for the peripheral nerves and are likely the cause of neuropathy, a cardinal manifestation of the disease. WHO recommends a protocol of multidrug therapy (MDT), which effectively controls the disease, hence contributing to the global elimination program. Early detection of leprosy and treatment by MDT are the most important steps in preventing deformity and disability. PMID- 12639459 TI - Herpes simplex virus (HSV)-associated erythema multiforme (HAEM): a viral disease with an autoimmune component. AB - Erythema multiforme (EM) is a clinical conundrum the name of which reflects the broad morphological spectrum of the lesions. Molecular and immunologic evidence that herpes simplex virus (HSV) causes a subset of EM lesions [herpes-associated EM (HAEM)] is reviewed, and new data are presented which suggest that autoreactive T-cells triggered by virus infection play an important role in HAEM pathogenesis. Disease development begins with viral DNA fragmentation and the transport of the DNA fragments to distant skin sites by peripheral blood mononuclear cells (PBMCs). HSV genes within DNA fragments deposited on the skin [notably DNA polymerase (Pol)] are expressed, leading to recruitment of HSV specific CD4+ Th1 cells that respond to viral antigens with production of interferon-gamma (IFN-gamma). This step initiates an inflammatory cascade that includes expression of IFN-gamma induced genes, increased sequestration of circulating leukocytes, monocytes and natural killer (NK) cells, and the recruitment of autoreactive T-cells generated by molecular mimicry or the release of cellular antigens from lysed cells. The PBMCs that pick up the HSV DNA [viz. macrophages or CD34+ Langerhans cells (LC) precursors], their ability to process it, the viral proteins expressed in the skin and the presence of epitopes shared with cellular proteins may determine whether a specific HSV episode is followed by HAEM development. Drug-associated EM (DIEM) is a mechanistically distinct EM subset that involves expression of tumor necrosis factor alpha (TNF-alpha) in lesional skin. It is our thesis that the polymerase chain reaction (PCR) assay for HSV DNA detection in lesional skin and staining with antibodies to IFN-gamma and TNF-alpha, are important criteria for the diagnosis of skin eruptions and improved patient management. PMID- 12639460 TI - A systematic review of adverse effects associated with topical treatments for psoriasis. AB - Mild to moderate psoriasis is a disease that can often be treated with topical medications. The diversity of topical therapies and their disparate side effects complicates treatment planning. Our purpose is to compare the rates of adverse events associated with different topical psoriasis treatments. A review of medical literature from 1996 to March, 2002 was conducted using guidelines set by QUORUM statement criteria. In monotherapy studies, corticosteriods caused fewer adverse reactions compared to vitamin D analogues and tazarotene. In combination studies adverse event rates were higher than in monotherapy studies, except for the combination of topical steroid and calcipotriene which decreased irritation. Irritant contact dermatitis was the main side effect with vitamin D analogues, tazarotene, dithranol or coal tar, while side effects of topical corticosteriods included headache, viral infection and skin atrophy. Topical agents for psoriasis are usually well-tolerated without severe side effects. Formulating a patient's medication regimen should take into account the needs for short-term management and long-term control of psoriasis. Since clearance is not a realistic expectation, reasonable goals should be set as excessive use of topical treatments may increase the risk of both cutaneous and systemic side effects. PMID- 12639461 TI - Cutaneous reactions to temporary tattoos. AB - While allergic reactions to henna painting are extremely rare, we have witnessed an increasing number of reports of cutaneous reactions after henna tattooing, also called temporary tattooing. The reason for this lies in the contemporary composition of the paint, which contains paraphenylenediamine (PPD). We present six patients who developed allergic contact dermatitis after skin painting with "black henna", and who showed hypersensitivity to PPD. We discuss the issue of the short sensitization period of our patients and the reported ones. PMID- 12639463 TI - Intense pulsed light technology and its improvement on skin aging from the patients' perspective using photorejuvenation parameters. AB - Intense pulsed light technology (IPL) has been used for photorejuvenation or the reversal of skin aging. There are few studies to address the putative benefits from the patients' perspective. This information is critical for adequate patient counselling and satisfaction. The goal of this study was to quantify the change in apparent facial age that can be expected from the patients' point of view. Using currently available parameters that have been individualized to patients' responses, five patients underwent treatment with IPL. Their subjective improvement scores were collected using visual analog scales. All patients showed improvement from their perspective. Quantification of this improvement showed that in this population, patients can expect, on average, a two year reduction in perceived age per treatment. This finding may be useful when discussing intense pulsed light technology with current patients and those prospective patients considering this procedure. PMID- 12639462 TI - Pilot study of a novel treatment for androgenetic alopecia using enriched cell culture medium: clinical trials. AB - Androgenetic Alopecia (AA) afflicts a large part of the population and of the many treatments available today none is completely satisfactory. Testing the efficacy and safety of a novel topical treatment for AA which is based on cell culture medium supplemented with insulin, thyroxin and growth hormone (CCM). The 48 participants classified as androgenetic alopecia Type II, III or IV on the Hamilton scale, concluded a randomized, vehicle-controlled, double-blind trial of 6 months duration. Under occlusive cover the gel was self applied for at least 3 hours daily. Evaluation was based on hair counts, investigator global assessment and participants self-administered questionnaire. Cessation of hair loss was reported by most participants within 28 weeks, and further confirmed by the hair count (HC) in ~80% of participants. Moreover, as early as 4 months after the start of the treatment, a time dependent increase of up to 50% in HC was observed. The average change in HC between the two groups differed significantly (p=0.007), with values of 4.1% for control and 13.8% for CCM. Following 4 months of treatment, a time dependent increase in HC (>10%) above minimal was observed in 55% of the CCM and 25% of the control and this trend continued. At 6 months 63% of the CCM and 33% of the control group exhibited increase of HC higher than 10%. The average increase in HC in the CCM and the control groups was 17.1% and 8.9% respectively (p=0.035). Self evaluation questionnaires revealed a time dependent increase in satisfaction in the CCMusers compared to the control. While the average score at T2 was similar in CCM and control (2.7 and 2.6 respectively), the score at T6 in the CCM increased to 5.9 and decreased to -0.4 in the control (p=0.007). Global-clinical evaluation following six months treatment revealed significantly (p=0.02) more hair loss in the control group (40%) compared to the CCM (7%) treated group. CCM was found effective in treating androgenetic alopecia in men. It induced cessation of hair loss, increased rate of hair growth and appearance of new hair. No side effects were reported or observed. PMID- 12639464 TI - Trichophyton raubitschekii: a new agent of dermatophytosis in Brazil? AB - A microbiological and ultrastructural study of Trichophyton raubitschekii recovered from a patient with tinea faciei is presented. This is the second case of isolation of this fungus in Sao Paulo and the sixth case in Brazil. Upon culture, the morphological pattern and the physiological tests performed confirmed the identification of T. raubitschekii. The ultrastructural study of T. raubitschekii showed the presence of a membrane-like structure located in the outer portion of the hyphal walls. This structure was bi-stratified and very like the one observed in T. rubrum. Is T. raubitschekii a new fungus emerging in Brazil or is it a T. rubrum variant? PMID- 12639466 TI - A case of dissecting cellulitis and a review of the literature. AB - Dissecting cellulitis (also called perifolliculitis capitis abscedens et suffodiens) manifests with perifollicular pustules, nodules, abscesses and sinuses that evolve into scarring alopecia. It predominantly occurs in African American men between 20-40 years of age, but can occasionally affect other races and women too. Associated musculoskeletal findings are sometimes reported. When it occurs with acne conglobata, hidradenitis suppurativa, and pilonidal cysts, the syndrome is referred to as the follicular occlusion triad or tetrad. Its course is chronic and relapsing, and treatment is often difficult. Medical therapies include isotretinoin, antibiotics, and prednisone. Destructive therapies include X-ray therapy, surgical excision, and skin grafting. Laser epilation of hair follicles is a promising new therapy for dissecting cellulitis. PMID- 12639465 TI - Tubular apocrine adenoma in association with syringocystadenoma papilliferum. AB - Tubular apocrine adenoma is a very rare sweat gland tumor. In this report, a case of tubular apocrine adenoma in association with syringocystadenoma papilliferum on the scalp is presented. The stroma of the tubular apocrine adenoma consisted of numerous, young fibroblasts with mitotic activity. It was difficult to distinguish stromal cells and epithelial cells from each other in some areas. The characteristics and differences in histopathologic and immunohistochemical findings in these tumors are described. PMID- 12639467 TI - Nodular secondary syphilis. AB - Secondary syphilis can have protean clinical manifestations and may present with unusual lesions, which may go unrecognized. We report a case of secondary syphilis with nodular lesions. A 22 year old male presented with nodular and annular skin lesions over the face, back and limbs and condylomata lata lesion at the penoscrotal junction associated with generalized lymphadenopathy, fever and malaise. Prior to onset of these lesions the patient also had history of a painless genital sore, which healed within two weeks. The serology revealed a reactive VDRL(1:64) and positive TPHA. The HIV serology was non-reactive. The patient responded to a single dose of benzathine penicillin, 2.4 million units, given intramuscularly. This case highlights that secondary syphilis may present with nodular lesions and should be suspected in the appropriate clinical setting. PMID- 12639468 TI - Metastatic cutaneous squamous cell carcinoma arising from a previous area of chronic hypertrophic lichen planus. AB - Malignant transformation of cutaneous lichen planus is a rare event. We report a 34 year old Caucasian male who presented with an exophytic tumor on the right foreleg. The tumor gradually developed within previous areas of histologically proven hypertrophic lichen planus that had existed for about 10 years. However, the current histological examination of the excised tumor revealed highly differentiated squamous cell carcinoma with a depth of tumor invasion of 10 mm. At that time, neither sentinel lymph node biopsy nor further imaging diagnostics revealed evidence for metastatic spreading. Nevertheless, five months after surgery inguinal lymph node metastases were detected. Initial chemotherapy and inguinal lymph node dissection were unable to stop the spread of the tumor. One year later, parailiacal lymph node metastases were detected by computed tomography. Further cycles of chemotherapy resulted in significant reduction of the parailiacal tumor masses. This report indicates that the long-standing hypertrophic form of lichen planus seems to have a considerable propensity for malignant transformation, even in young patients. PMID- 12639469 TI - Annular atrophic plaques of skin (Christianson's disease). AB - Atrophic plaques with white borders are occasionally seen on sun-exposed areas of the skin. These patients are usually elderly and have solar elastosis. This condition is referred to as annular atrophic plaques of skin and we describe a typical case. PMID- 12639470 TI - Acquired Blaschko Dermatitis-seventh case. AB - The February edition of Dermatology Online Journal 2002;7(1):8, contained an article entitled Acquired Blaschko Dermatitis.[1] A 64 year old patient with erythematous patches and papules in a reticulate pattern on the left upper extremity and on the left side of the chest, abdomen, back and buttock was described. Three months later, in the Department of Dermatology, Health Center Krusevac, we examined a 65 year old woman with similar lesions, distributed in a linear pattern on her right lower limb, following the lines of Blaschko. PMID- 12639471 TI - Cutaneous variant of angiokeratoma corporis diffusum. AB - A case of angiokeratoma corporis diffusum (ACD) involving the skin of a 22-year old patient presenting with normal physical and mental development is reported. ACD presenting with skin lesions alone is a rare but specific clinical entity, which differs from hereditary sphingolipidoses such as Fabry's disease. PMID- 12639472 TI - Methotrexate-induced acral erythema with bullous reaction. AB - Chemotherapy-induced acral erythema (CIAE), a toxic reaction to a number of different chomotherapeutic agents, causes a symmetrical, painful erythema of both the palms and soles which is self-limiting. The association of this syndrome with methotrexate is unusual; only nine cases have been reported in the literature. We describe the tenth case of this syndrome associated with methotrexate, which is also the third case of the bullous variant of methotrexate-induced acral erythema. Our case is unusual in that the acral erythema was present only on the soles of the feet and in that it was associated with the presence of diffuse maculopapular lesions over the legs and trunk. PMID- 12639473 TI - Report of a new case with four skin diseases. AB - A 70 year old man is presented with four distinct skin diseases that were diagnosed concomitantly. Lichen planus, vitiligo, lichen sclerosis, and disseminated actinic porokeratosis were all diagnosed clinically and histologically in this man, who was otherwise healthy. PMID- 12639474 TI - Cicatricial alopecia due to sarcoidosis. AB - Sarcoidosis of the scalp is a rare manifestation of cutaneous sarcoidosis. We report a case in a Syrian man without systemic involvement. Sarcoidosis should be considered in the differential diagnosis of cicatricial and non-cicatricial alopecia along with discoid lupus erythematous (DLE), lichen planopilaris, and scleroderma. PMID- 12639475 TI - Easy photodermoscopy for teledermatology. PMID- 12639476 TI - Offering rubella vaccination to sexually active adolescents. PMID- 12639477 TI - Pneumococcal conjugate vaccines: proceedings from an interactive symposium at the 41st Interscience Conference on Antimicrobial Agents and Chemotherapy. AB - Globally, Streptococcus pneumoniae is a leading cause of invasive and noninvasive disease in infants and young children. The emergence of antibiotic-resistant strains has increased interest in prevention through immunization. Currently, the only available conjugate pneumococcal vaccine is a seven-valent formulation, PNCRM7. This paper presents excerpts from a symposium that provided an update of ongoing surveillance data and clinical trials evaluating pneumococcal conjugate vaccines. The topics addressed included: (1) PNCRM7 postmarketing safety data; (2) the impact of PNCRM7 in premature infants; (3) the direct and indirect effect of pneumococcal conjugate vaccines on colonization; (4) the effect of pneumococcal conjugate vaccines on replacement disease and the rate of resistance among replacement serotypes; (5) the current recommendations for the use of PNCRM7; and (6) the potential impact of conjugate vaccines in Europe and the Asia Pacific region. PMID- 12639478 TI - Cross-protective and infection-enhancing immunity in mice vaccinated against flaviviruses belonging to the Japanese encephalitis virus serocomplex. AB - The Japanese encephalitis virus serocomplex is a group of mosquito-borne flaviviruses that cause severe encephalitic disease in humans. The recent emergence of several members of this serocomplex in geographic regions where other closely related flaviviruses are endemic has raised urgent human health issues. Thus, the impact of vaccination against one of these neurotropic virus on the outcome of infection with a second, serologically related virus is unknown. We show here that immunity against Murray Valley encephalitis virus in vaccinated mice can cross-protect but also augment disease severity following challenge with Japanese encephalitis virus. Immunepotentiation of heterologous flavivirus disease was apparent in animals immunized with a 'killed' virus preparation when humoral antiviral immunity of low magnitude was elicited. PMID- 12639480 TI - A fish specific expression vector containing the interferon regulatory factor 1A (IRF1A) promoter for genetic immunization of fish. AB - DNA plasmid vectors were constructed with inducible rainbow trout promoters to take the place of the cytomegalovirus immediate early promoter in DNA vaccines for fish. DNA fragments containing the promoter regions upstream of the rainbow trout Mx1 and interferon regulatory factor 1A (IRF1A) genes were obtained and their nucleotide sequences were determined. Vectors containing the G gene of IHNV linked to the trout promoters were found to be very effective as vaccines in rainbow trout. In addition, we demonstrated that the IRF1A promoter is transcriptionally active in mouse cells NIH-3T3 suggesting that this fish promoter might be used for mammalian DNA vaccine development as well. PMID- 12639479 TI - Synthetic IgE peptide vaccine for immunotherapy of allergy. AB - An immunotherapeutic vaccine for allergy was produced by designing IgE-based synthetic peptide immunogens and selecting them for functional immunogenicity. The vaccine targets the binding site on IgE for the high affinity receptor Fc epsilon RI, by active immunization. The peptide target site on IgE heavy chain was selected from among the amino acid sequences for the C epsilon 2, C epsilon 3, and C epsilon 4 domains. These were characterised by epitope mapping studies for cross-reactivity to IgE and functional antigenicity. A peptide, modified from positions 413-435 of a loop region of C epsilon 3 and subjected to conformational constraint, elicited anti-IgE antibodies that blocked IgE-mediated histamine release. It was immunopotentiated by linkage to a promiscuous T helper site to produce a wholly synthetic chimaeric immunogen. This immunogen was shown to induce polyclonal site-specific anti-IgE antibodies that obstruct binding to Fc epsilon RI, inhibit histamine release by IgE-sensitised basophils, inhibit passive cutaneous anaphylaxis, and do not signal degranulation. Immunized dogs experienced significant reductions in total serum IgE. PMID- 12639481 TI - Monoclonal anti-idiotypic antibody mimicking the gastrointestinal carcinoma associated epitope CO17-1A elicits antigen-specific humoral and cellular immune responses in colorectal cancer patients. AB - Monoclonal rat anti-idiotypic antibody (Ab2) BR3E4 mimicking the colorectal carcinoma (CRC)-associated epitope CO17-1A induced antigen-specific humoral and cellular immune responses in mice and rabbits. Ab2 BR3E4 was administered in a phase I trial to CRC patients either as intact IgG or as F(ab')(2) coupled to keyhole limpet hemocyanin (KLH). There was a trend for the F(ab')(2)-KLH immunized patients to show higher immune response rates (18/21 and 5/15 patients with anti-anti-idiotypic antibodies and T cells, respectively) than the IgG immunized patients (15/23 and 3/15 patients positive). Clinical responses were rare in these patients with liver metastases. PMID- 12639482 TI - Induction of cytotoxic T lymphocyte responses by cholera toxin-treated bone marrow-derived dendritic cells. AB - Cholera toxin (CT), a powerful mucosal adjuvant, is a potent inducer of Th2-type responses via activation of co-stimulatory molecules for the induction of IgA antibody responses. Less appreciated is the ability of CT to induce and regulate cytotoxic T lymphocyte (CTL) responses. In order to help for clarifying mechanisms underlying the CTL-inducing ability of CT, we have examined the effects of CT on dendritic cells (DCs) that could lead to the induction of cytotoxic CD8(+) T cells. When bone marrow-derived DCs (BM-DCs) were cultured with CT in vitro, B7-1 but not B7-2 molecules were significantly enhanced and allogenic CTL responses were induced. Also, increased numbers of IFN-gamma secreting CD8(+) T cells were elicited when CT-treated BM-DCs were co-cultured with allogenic CD8(+) CTLs. Antibody blockade of B7-1 on CT-treated BM-DCs suppressed allogenic CTL responses, further indicating the importance of CT induced B7-1 molecules on DCs for the acquisition of cytolytic function by CTL precursors. CD40 signaling was proven not necessary for the CT-induced CTL response since CT-treated CD40(-/-) BM-DCs developed CTL responses equivalent to those detected in CT-treated BM-DCs derived from normal mice. Our results suggest that CT-treated DCs are effective inducers of CD8(+) CTL, and this induction is mediated through CT's ability to enhance B7-1 expression on DCs. PMID- 12639483 TI - Comprehensive systematic surveillance for adverse effects of anthrax vaccine adsorbed, US Armed Forces, 1998-2000. AB - Routine vaccinations of US military personnel with Anthrax Vaccine Adsorbed began in 1998. To systematically identify clinical diagnoses reported more frequently after vaccination than before, all military personnel were retrospectively assigned to pre- or post-vaccination cohorts. Cohort assignments were based on vaccination statuses each day of the 3-year surveillance period. For each cohort, rates of hospitalizations and ambulatory visits for 843 specific diagnoses were calculated using data in a public health surveillance system. Compared to the pre vaccination cohort, the post-vaccination cohort had statistically higher rates of hospitalizations for 17 diagnoses, of ambulatory visits for 34 diagnoses, and in both clinical settings for one diagnosis (malaria). After accounting for systematic differences in coding/reporting and residual confounding, the number and nature of clinical diagnoses more frequent after anthrax vaccination than before were consistent with expectations due to random variation. This surveillance suggests that Anthrax Vaccine Adsorbed has few, if any, clinically significant adverse effects. PMID- 12639484 TI - Enhanced induction of hepatitis C virus-specific cytotoxic T lymphocytes and protective efficacy in mice by DNA vaccination followed by adenovirus boosting in combination with the interleukin-12 expression plasmid. AB - We evaluated the prime-boost immunization consisting of hepatitis C virus (HCV) core expression plasmid (pCEP4-core) and replication-defective adenovirus expressing HCV-core (Adex1SR3ST) for core-specific CTL induction in mice. Compared to a single booster, double boosters after priming enhance CTL induction. The prime-double boosts immunization involving pCEP4-core priming followed by pCEP4-core and Adex1SR3ST boostings (pC/pC/aC) can induce core specific CTLs as well as other combinations: pC/aC/aC; aC/pC/pC; aC/aC/aC, whereas pC/pC/pC does not induce CTLs. Furthermore, co-administration of interleukin-12 (IL-12) expression plasmid leads to the highly efficient CTL induction and clearance of HCV-core expressing vaccinia virus challenged. Thus, the prime-double boosts immunization together with IL-12 may be promising for HCV vaccine. PMID- 12639485 TI - Modulating gene expression using DNA vaccines with different 3'-UTRs influences antibody titer, seroconversion and cytokine profiles. AB - To determine if modulating the amount of foreign antigen produced by a DNA vaccine can influence the overall intensity and cytokine polarization of the ensuing immune response, three different plasmids, each encoding the hepatitis B (HB) surface antigen, were constructed. In each construct, HBs gene expression was driven by the cytomegalovirus immediate early promoter, but differed in the 3'-untranslated regions (3'-UTR) containing the polyadenylation sequence. These 3'-UTR sequences were derived from either the hepatitis B virus (HBVpA), bovine growth hormone (BGHpA), or rabbit beta-globin (betapA). BALB/c mice were immunized intramuscularly with equimolar amounts of each plasmid and blood was collected bi-weekly. Following immunization, total IgG titers correlated with in vitro antigen production levels (from transfected CHO cells), as evidenced by the following response pattern: HBVpA>BGHpA>>betapA. All groups demonstrated a heavy bias toward a Th1 immune response, as evidenced by high serum IgG2a/IgG1 ratios and the predominance of IFN-gamma over IL-4 secretion from cultured splenocytes. In addition, the HBVpA construct resulted in a seroconversion rate of 100%, in comparison to 40-50% in the BGHpA, and 0% in the betapA group. Surprisingly, splenocytes isolated from mice immunized with the betapA construct secreted the highest levels of IFN-gamma. Taken together, these findings suggest that altering the level of gene expression not only affects the overall titer and seroconversion rates of vaccinated animals, but also may play a role in modulating cytokine profiles. PMID- 12639487 TI - Issues in selection of DTwP-based combination vaccines. AB - This study examines regulatory, supply, acceptance and financing issues of combinations based on diphtheria-tetanus-pertussis (whole cell) vaccines (DTwP). These combination vaccines are currently produced in Europe mostly for export. Future regulatory oversight issues could have an impact on their availability. Before use of acellular pertussis-containing vaccines, the number of doses of DTwP vaccines offered in response to United Nations agency procurement tenders far exceeded the projected demand. Current demand and supply are converging. Most of that supply comes from developing country manufacturers, a potential new source of combination vaccines as well. The expected development and use of DTwP based combination vaccines raises antigen allocation issues that could further affect supply. These combination vaccines have strong programmatic advantages, but pose complex selection issues involving disease burden, presentation, and availability of long-term financing. Vaccine price is not the major driving factor. A model examining important selection factors for regional country groupings provides predictions that have been validated by decisions on use of DTwP-based combination vaccines. This model may be useful in providing long-term uptake projections for other combination vaccines. PMID- 12639486 TI - Expression of malaria transmission-blocking vaccine antigen Pfs25 in Pichia pastoris for use in human clinical trials. AB - In previously published studies, Saccharomyces cerevisiae recombinant protein expression systems have been employed to express the malaria parasite antigen Pfs25, a candidate transmission-blocking vaccine antigen against Plasmodium falciparum malaria. However, despite having been in two Phase 1 trials, the recombinant Pfs25 so produced (previously called TBV25H) exists as a mixture of two monomeric protein conformational forms, Pfs25H-A and Pfs25H-B. In this study, we optimized the expression and purification of the two Pfs25H conformers in S. cerevisiae, and characterized their biochemical and antigenic properties, immunogenicities, and transmission-blocking activities. Pfs25H-A is apparently homogeneous, and has the correct conformation as measured by monoclonal antibody recognition. It is, however, expressed at a low yield of only 0.19mg/l. By contrast, Pfs25H-B is produced as a heterogeneous population of molecules that do not seem to have the correct conformation. Nonetheless, both forms appear equally effective in their ability to produce transmission-blocking antibodies in mice. To address the low yield seen with S. cerevisiae, we also expressed Pfs25 in Pichia pastoris. P. pastoris is apparently superior to S. cerevisiae in producing higher yield, immunologically more potent, biologically more active Pfs25H-A. PMID- 12639488 TI - Immunization onto shaved skin with a bacterial enterotoxin adjuvant protects mice against respiratory syncytial virus (RSV). AB - This study evaluated the potential of the skin as a non invasive route for RSV vaccination using two G protein-derived molecules, G2Na and G5 in mice. G2Na contains T and B-cell epitopes whether G5 is a pure B-cell epitope. In contrast to G5, G2Na coadministered with CT three times at 1 month interval onto 1cm of square area shaved skin, elicited a consistent serum anti-G2Na and anti-CT IgG response. The anti-G2Na IgG response was dominated by IgG1 isotype, an indirect marker of a Th2 type of response. Dramatic reduction and decrease of RSV titers in lung tissues and in the nasal tract, respectively, following intranasal virus challenge revealed biological relevance of the transcutaneous immunization in the context of RSV vaccine. These results suggest that the transcutaneous route may offer a promising potential for novel RSV vaccine strategy, simple, painless and economical. PMID- 12639489 TI - Immune response and protection elicited by DNA immunisation against Taenia cysticercosis. AB - The study evaluated DNA vaccination in Taenia solium cysticercosis prevention by using cDNA of an antigen (cC1) from T. solium metacestode. pcDNA3-cC1 DNA vaccine was constructed by inserting the cDNA into the eukaryotic expression plasmid pcDNA3. Positive expression of the pcDNA3-cC1 product was confirmed by its transfection into COS7 cell and enzyme-linked immunoabsorbent assay using serum of pigs infected with T. solium metacestode. Immunisation of BALB/c mice with three injections of pcDNA3-cC1 induced antigen-specific immune responses of the Th1 phenotype. Inoculation of new-born pigs induced protection against challenge with T. solium by 73.3% reduction of the metacestode number. Antibodies elicited by DNA immunisation with pcDNA3-cC1 specifically reacted with native cC1 protein, which was mainly restricted to the cyst wall of T. solium metacestode. Positive apoptosis signals were also detected in the cyst wall cells of metacestode slices from pigs immunised with pcDNA3-cC1 by TUNEL staining method. Those suggested that apoptosis played a role in protecting pigs immunised with pcDNA3-cC1 nucleic acid vaccine from pathogen challenge. PMID- 12639490 TI - The vaccine data link in Nha Trang, Vietnam: a progress report on the implementation of a database to detect adverse events related to vaccinations. AB - Real, perceived and unknown adverse events secondary to vaccinations are a source of concern for care providers of children. In the USA large linked databases have provided helpful information regarding the safety of vaccines. Very little prospectively collected data on vaccine safety is available from resource poor countries, but safety concerns may be even more relevant in such settings. Vaccine manufacturers do not have to pass the same rigorous safety standards as vaccine manufacturers in rich countries. Vaccines, which protect against cholera, Japanese encephalitis, rabies or typhoid fever are predominantly used in resource poor, tropical countries and frequently do not undergo vigorous post marketing surveillance. New vaccines specifically suited for resource poor countries are sometimes marketed without the scrutiny of vigilant, independent regulatory authorities. We describe here the design and implementation of a large linked database for a semi-rural province in central Vietnam. The design overcomes several problems inherent in data bases of medical events and vaccinations in developing countries. Assigning a permanent identification (ID) number to each resident avoids the ambiguities of ID numbers based on the address. The distribution and use of medical identification cards with a permanent ID number assists in the unambiguous identification of vaccinees and patients. Medical records of all admissions are coded according to International Classification of Diseases (ICD-10) and transcribed into a computer system. Because these processes are novel the data collected by the study will be validated. Project staff will check records on vaccinations and hospital admissions through household visits at regular intervals. Data describing vaccinations and medical events are linked to the data collected by the project staff in a computer system. Based on the validation of the data we hope to optimize this model. Once we find the model working it is planned export this vaccine data safety link to other settings of similar economic status. PMID- 12639491 TI - Boosting immunity to influenza H5N1 with MF59-adjuvanted H5N3 A/Duck/Singapore/97 vaccine in a primed human population. AB - In 1997, influenza A/Hong Kong/97 (H5N1) emerged as a potential human threat. In 1999, a randomised study comparing two doses of MF59-adjuvanted and non adjuvanted influenza A/Duck/Singapore/97 (H5N3) surface-antigen vaccine found non adjuvanted vaccine was poorly immunogenic. Addition of MF59 significantly boosted antibody to H5N1 to levels associated with protection. At 16 months, we undertook a follow-up study to assess the effect of H5N3 revaccination. Geometric mean titres (GMTs) of antibody by haemagglutination-inhibition (HI), microneutralisation (MN) and single radial haemolysis (SRH) indicated that protective antibody titres did not exist at 16 months after two-dose priming. Twenty-one days after revaccination, there was significant boosting of antibody compared to GMTs achieved 21 days after two-dose priming in the original study (P<0.001). MF59 significantly increased GMTs of antibody when compared to non adjuvanted vaccine (P<0.001). PMID- 12639492 TI - Recombinant Vibrio cholerae ghosts as a delivery vehicle for vaccinating against Chlamydia trachomatis. AB - An efficacious vaccine is needed to control the morbidity and burden of rising healthcare costs associated with genital Chlamydia trachomatis infection. Despite considerable efforts, the development of reliable chlamydial vaccines using conventional strategies has proven to be elusive. The 40kDa major outer membrane protein (MOMP) of C. trachomatis is so far the most promising candidate for a subunit vaccine. The lack of satisfactory protective immunity with MOMP-based vaccine regimens to date would suggest that either MOMP alone is inadequate as a vaccine candidate or better delivery systems are needed to optimize the effect of MOMP. Recombinant Vibrio cholerae ghosts (rVCG) are attractive for use as non living vaccines because they possess strong adjuvant properties and are excellent vehicles for delivery of antigens of vaccine relevance to mucosal sites. The suitability of the ghost technology for designing an anti-chlamydial vaccine was evaluated by constructing a rVCG vector-based candidate vaccine expressing MOMP (rVCG-MOMP) and assessing vaccine efficacy in a murine model of C. trachomatis genital infection. Intramuscular delivery of the rVCG-MOMP vaccine induced elevated local genital mucosal as well as systemic Th1 responses. In addition, immune T cells from immunized mice could transfer partial protection against a C. trachomatis genital challenge to nai;ve mice. These results suggest that rVCG expressing chlamydial proteins may constitute a suitable subunit vaccine for inducing an efficient mucosal T cell response that protects against C. trachomatis infection. Altogether, the potency and relatively low production cost of rVCG offer a significant technical advantage as a chlamydial vaccine. PMID- 12639493 TI - A population-based register study of vaccine coverage among children in Greenland. AB - To assess coverage rates of standard childhood vaccinations in Greenland, a geographically isolated and sparsely populated area, and to determine risk factors for low coverage, we performed a register-based cohort study among 596 children from 1993 to 1998 in Sisimiut, Greenland. For vaccines given before the age of 2 years (pertussis 1-3, DT-IPV 1-3, and MMR 1) coverage rates in general were impressively high being on or above levels of Western countries. A risk factor for low coverage was migration. The Greenlandic vaccination model with free vaccinations administered by health workers who systematically call in children at scheduled times seems highly efficient and could be a model for other similar countries. PMID- 12639494 TI - Serological response and poliovirus excretion following different combined oral and inactivated poliovirus vaccines immunization schedules. AB - A controlled study was conducted in Karachi, Pakistan to compare humoral and mucosal immune responses against polioviruses in infants who received oral poliovirus vaccine (OPV) at birth and at 6, 10, and 14 weeks according to the Expanded Program on Immunization (EPI) with infants who received either three doses of inactivated poliovirus vaccine (IPV) at 6, 10, and 14 weeks together with OPV or one additional dose of IPV at 14 weeks together, with the last dose of OPV. A total of 1429 infants were enrolled; 24-week serum specimens were available for 898 infants (63%). They all received a challenge dose of OPV type 3 at 24 weeks of age. The addition of three doses of IPV to three doses of OPV induced a significantly higher percentage of seropositive children at 24 weeks of age for polio 1 (97% versus 89%, P<0.001) and polio 3 (98% versus 92%) compared to the EPI schedule. However, the one supplemental dose of IPV at 14 weeks did not increase the serological response at 24 weeks. Intestinal immunity against the challenge dose was similar in the three groups. Combined schedules of OPV and IPV in the form of diphtheria-pertussis-tetanus-IPV vaccine (DPT-IPV) may be useful to accelerate eradication of polio in developing countries. PMID- 12639495 TI - Design of the Dutch prevention of influenza, surveillance and management (PRISMA) study. AB - Rationale and design of a study on the cost-effectiveness of the Dutch influenza vaccination campaign are described. During two influenza epidemics, about 75,000 primary care patients recommended for influenza vaccination are included. Cases have fatal or non-fatal influenza, pneumonia, otitis media, acute respiratory disease (ARD), heart failure, myocardial infarction, depression or diabetes dysregulation. Per case four controls are sampled, frequency matched on age and high-risk co-morbidity (<18 years, 18-64, >/=65 healthy, >/=65 with co morbidity). Baseline and outcome data are retrieved from patient records. During the 1999-2000 influenza A epidemic 5891 (7.9%) high-risk children, 24,848 (33.2%) high-risk adults aged 18-64 years, 18,484 (24.7%) elderly with co-morbidity and 25,527 (34.1%) healthy elderly had been included. The mortality rate was 5.2 per 1000 and 2035 non-fatal outcome events were recorded (incidence rate 27.2/1000). PMID- 12639496 TI - Coproantibodies to the enteropathogenic Escherichia coli vaccine candidates BfpA and EspB in breastfed and artificially fed children. AB - A diagnostic test was developed to discriminate active from past enteropathogenic Escherichia coli (EPEC) infection, which uses the affinity-purified recombinant proteins BfpA (bundle-forming pilus (BFP) structural repeating subunit A) and EspB (pore-forming secreted protein B) as reliable markers of virulence to detect antigen-specific coproantibodies by immunoblot analysis, and verification of active typical EPEC infection by gene-specific (bfpA and espB) PCR amplification using DNA extracted directly from specimens and/or culture-enriched preparations. To begin addressing the potential protective role of anti-EPEC antibodies at early age, the prevalence of IgA coproantibodies to these antigens was determined in either breastfed or artificially fed children <2 years of age hospitalized for watery diarrhea. PMID- 12639497 TI - DNA vaccination of neonate piglets in the face of maternal immunity induces humoral memory and protection against a virulent pseudorabies virus challenge. AB - DNA vaccination represents a unique opportunity to overcome the limitations of conventional vaccine strategy in early life in the face of maternal-derived immunity. We used the model of pseudorabies virus (PRV) infection in pigs to further explore the potential of DNA vaccination in piglets born to sows repeatedly vaccinated with a PRV inactivated vaccine. A single immunisation of 8 week-old piglets with a DNA vaccine expressing secreted forms of PRV gB, gC, and gD, triggered an active serological response, confirming that DNA vaccination can over-ride significant residual maternal-derived immunity. A clear anamnestic response was evidenced when a secondary DNA vaccination was performed at 11 weeks of age, suggesting that DNA vaccination, performed in the face of passive immunity, elicited a strong humoral memory. We subsequently explored the potential of DNA vaccination in neonate piglets (5-6 days of age) in the face of very high titres of maternal antibodies and demonstrated that very high titres of passive antibodies selectively inhibited serological responses but not the establishment of potent memory responses. Finally, we demonstrated that DNA vaccination provided protection against an infectious PRV challenge at the end of the fattening period (i.e. at approximately 5 months of age). Collectively, our results pave the way for a new flexible vaccination program, which could ensure uninterrupted protection of fattening pigs over their entire economical life under field conditions. PMID- 12639498 TI - Negatively charged 2- and 10-microm particles activate vanilloid receptors, increase cAMP, and induce cytokine release. AB - Exposure to airborne pollutants, such as particulate matter (PM), is associated with increased mortality and morbidity. Indirect evidence suggested that PM induced responses could be initiated by the activation of proton-gated receptors, including vanilloid receptors (VRs) and acid-sensitive ion channels (e.g. ASICS). We tested this hypothesis by characterizing the effects of 10- and 2-microm polystyrene carboxylate-modified particles (PC(10) and PC(2)) on HEK 293 cells expressing VR1 receptors, rat trigeminal ganglion (TG) neurons, and BEAS-2B airway epithelial cells. Zeta potential measurements revealed that these particles are negatively charged, meaning that when they adhere to a membrane they can lower the surface pH and activate proton-gated receptors. Both types of PCs induced currents and/or elevated intracellular Ca(2+) in cells that were capsaicin sensitive (CS). In about 70% of CS neurons, 10 microM capsazepine (CPZ), a VR antagonist, blocked PC-induced responses. In TG neurons in which VRs were blocked or desensitized, PCs induced an amiloride-inhibitable inward current having the characteristics of ASIC-mediated currents. Incubation of TG neurons with either capsaicin or PCs produced a CPZ-sensitive increase in cyclic AMP and cytokine (IL-6) release. In summary, we provide unequivocal evidence demonstrating that negatively charged PCs could activate VR1 and other proton gated receptors. These data suggest that pharmacological manipulation of such receptors could prevent the physiological actions of PMs. PMID- 12639499 TI - Lead enters Rcho-1 trophoblastic cells by calcium transport mechanisms and complexes with cytosolic calcium-binding proteins. AB - Within the placenta, a specialized Ca(2+) transport pathway develops in trophoblasts to promote growth of the fetus and hypothetically to enhance fetal uptake of Pb(2+). This hypothesis could not be tested until a method to monitor Pb(2+) influx by indo-1 fluorescence quench became available. We have applied this new method to cultured undifferentiated and differentiated Rcho-1 trophoblastic cells. Pb(2+) concentrations of 1 and 10 microM are equivalent to blood levels of 20 and 200 microg/dl in pregnant women. Over this range, Pb(2+) uptake increased with time and concentration in medium containing 1 mM Ca(2+) but was greater in Ca(2+)-omitted solutions. Activation of capacitative Ca(2+) entry (CCE) with thapsigargin, an endoplasmic reticulum (ER) Ca(2+) pump inhibitor, increased Pb(2+) uptake, while inhibition of CCE by La(3+) decreased influx. Parathyroid hormone-related peptide (PTHrP) stimulates the synthesis of Ca(2+) binding proteins (CaBPs), as well as Ca(2+) transporters, during trophoblastic differentiation. Pretreatment for 72 h with PTHrP increased Pb(2+) uptake by undifferentiated Rcho-1 cells but had little effect on the quench in differentiated cells, probably due to their greater content of CaBPs which competed for Pb(2+)-binding with indo-1. This competition was most evident in differentiated cells when 1 microM Pb(2+) caused an initial quench, followed by a rise in fluorescence. This rise was not inhibited by thapsigargin, thereby ruling out sequestration into the ER and leaving complexation of Pb(2+) by CaBPs as the most plausible interpretation. We conclude that trophoblasts have the ability to clear Pb(2+) from the maternal circulation and deliver it to the fetus. PMID- 12639500 TI - Airway fibrosis in a mouse model of airway inflammation. AB - BALB/c mice were sensitized to ovalbumin by systemic injection and then exposed for up to 8 weeks to ovalbumin aerosols in whole body chambers. A pattern of airway inflammation, mucous cell hypertrophy and hyperplasia, and airway remodeling with submucosal fibrosis was observed as lesions evolved over time. Larger conducting airways were removed from the lungs by microdissection. Airway fibrosis was quantified by direct assay for collagen content, which was significantly increased after 4 and 8 weeks of exposure to ovalbumin aerosol. Based upon PCR analysis of mRNA levels in the airways, most of the newly synthesized collagen was Type I. Relaxin, administered by continuous infusion over the second half of a 4-week exposure to ovalbumin, was able to inhibit the accumulation of collagen in the airways of exposed mice. Thus, stimulation of collagen degradation by an activator of collagen breakdown by matrix metalloproteinases appears to be an effective therapeutic strategy in prevention of airway fibrosis in this animal model. Whole body plethysmography of unrestrained mice indicated functional changes in airway reactivity in the lungs of exposed animals occurring in conjunction with the reported structural changes. This result indicates that the ovalbumin-exposed mouse may be a suitable model for examining structure-function relationships in the lungs of animals with a predictable time course of airway inflammation, remodeling, and fibrosis and for testing potential new drugs for treatment of asthma or chronic bronchitis at a mechanistic level. PMID- 12639501 TI - Low-level arsenite causes accumulation of ubiquitinated proteins in rabbit renal cortical slices and HEK293 cells. AB - Arsenic is a known human carcinogen that affects a variety of processes within the cell. In this study, the effects of environmentally relevant As(III) exposures on the ubiquitin (Ub)-proteasome pathway have been investigated. Low level As(III) exposure (0.5 - 10 microM) causes an accumulation of high-molecular weight ubiquitin protein conjugates in both precision-cut rabbit renal-cortical slices and human embryonic kidney (HEK) 293 cells. The As(III) doses that induced these molecular changes were subcytotoxic in both model systems. Doses of 10 microM As(III) decreased cellular activity of the 20S proteasome by 40 and 15% in slices and HEK293 cells, respectively. As(III) did not cause any notable difference in Ub-conjugating activity of rabbit renal slices or HEK293 cells. Since ubiquitination plays such a vital role in maintaining cellular homeostasis, this noticeable perturbation of cellular ubiquitination is likely to have a multitude of signaling effects within the cells and may contribute to the pathogenesis of low-level arsenic. PMID- 12639502 TI - Neurotoxicity induced in differentiated SK-N-SH-SY5Y human neuroblastoma cells by organophosphorus compounds. AB - Organophosphorus (OP) compounds used as insecticides and chemical warfare agents are known to cause potent neurotoxic effects in humans and animals. Organophosphorus-induced delayed neuropathy (OPIDN) is currently thought to result from inhibition of neurotoxic esterase (NTE), but the actual molecular and cellular events leading to the development of OPIDN have not been characterized. This investigation examined the effects of OP compounds on the SY5Y human neuroblastoma cells at the cellular level to further characterize cellular targets of OP neurotoxicity. Mipafox and paraoxon were used as OP models that respectively do and do not induce OPIDN. Mipafox (0.05 mM) significantly decreased neurite length in SY5Y cells differentiated with nerve growth factor (NGF) while paraoxon at the same concentration had no effect when evaluated after each of three 4-day developmental windows during which cells were treated daily with OP or vehicle. In contrast, paraoxon but not mipafox altered intracellular calcium ion levels ([Ca(2+)](i)), as seen in three types of experiments. First, immediately following the addition of a single high concentration of OP to the culture, paraoxon caused a transient increase in [Ca(2+)](i), while mipafox up to 2 mM had no effect. Paraoxon hydrolysis products could also increase intracellular Ca(2+) levels, although the pattern of rise was different than it appeared immediately after paraoxon administration. Second, repeated low-level paraoxon treatment (0.05 mM/day for 4 days) decreased basal [Ca(2+)](i) in NGF differentiated cells, though mipafox had no effect. Third, carbachol, a muscarinic acetylcholine receptor agonist, transiently increased [Ca(2+)](i) in differentiated cells, an affect attenuated by 4-day pretreatment with paraoxon (0.05 mM/day), but not by pretreatment with mipafox. These results indicate that the decrease in neurite extension that resulted from mipafox treatment was not caused by a disruption of Ca(2+) homeostasis. The effects of OPs that cause or do not cause OPIDN were clearly distinguishable, not only by their effects on neurite length, but also by their effects on Ca(2+) homeostasis in differentiated SY5Y cells. PMID- 12639524 TI - CT colonography: methods, pathology and pitfalls. AB - Computed tomography colonography (CTC) is a relatively new technique that is currently challenging more established methods of large bowel imaging. Several workers have suggested CTC surpasses the barium enema and approaches conventional endoscopy for detection of colorectal neoplasia. Accurate diagnosis relies on technically good studies, the main aim of which is adequate bowel cleansing and distension. Furthermore, the learning curve is steep and normal colonic anatomy has to be re-learned in a CT context. This review aims to describe the technique, revise the imaging features of both normal and pathological colon, and to highlight potential diagnostic pitfalls and their avoidance. PMID- 12639503 TI - Genotyping and site-directed mutagenesis of a cytochrome P450 meander Pro-X-Arg motif critical to CYP4B1 catalysis. AB - CYP4B1 isoforms from rodents and other common laboratory animals are involved in the bioactivation of a range of protoxins, including 2-aminofluorene, 4 ipomeanol, and valproic acid. However, an earlier study provided evidence for a human allele encoding a nonfunctional CYP4B1 enzyme due to a Pro427Ser transversion in the meander region of the protein. In the present study, the CYP4B1 gene from several racial groups, Caucasians, African-Americans, and Hispanics, and from six nonhuman primate species was genotyped using a PCR-Hinf1 restriction enzyme fragment length polymorphism assay or by direct sequencing. All human populations examined were found to possess only the Ser allele at codon 427 ((1279)TCT) and all of the nonhuman primate species possessed only the Pro (CCT) allele. Therefore, an inactivating (1279)C-->T mutation in the human CYP4B1 gene likely arose following divergence of the Homo and Pan clades. Amino acid sequence alignments revealed further that this key Pro residue is located two amino acid residues N-terminal to the distal Arg of a Glu-Arg-Arg triad thought to participate in heme binding and/or redox partner interactions. Mutation of the corresponding Arg424 residue in rabbit CYP4B1 to Leu, but not His, resulted in a loss of lauric acid hydroxylase activity and ability to generate a reduced-CO binding spectrum. These data provide additional evidence for the importance of this meander region Pro-X-Arg motif in CYP4B1 heme binding and catalytic function. PMID- 12639525 TI - Unusual causes of colonic wall thickening on computed tomography. AB - Computed tomography (CT) appearances in "colitis" are often non-specific, and include mural thickening and mesenteric fat stranding. In the western world, the majority of cases will have, or be subsequently diagnosed with, inflammatory bowel disease, pseudomembranous colitis or ischaemic colitis. However, other rare conditions may also produce these rather non-specific signs. We present a number of cases demonstrating colonic wall thickening on CT due to rarer diagnoses, which are correlated with the histopathological features. Some of these CT appearances have not been described previously in the literature. PMID- 12639526 TI - Imaging in haematopoietic stem cell transplantation. AB - Haematopoietic stem cell transplantation (SCT) is used to treat a wide range of malignant and non-malignant haematological conditions, solid malignancies, and metabolic and autoimmune diseases. Although imaging has a limited role before SCT, it is important after transplantation when it may support the clinical diagnosis of a variety of complications. It may also be used to monitor the effect of therapy and to detect recurrence of the underlying disease if the transplant is unsuccessful. We present a pictorial review of the imaging of patients who have undergone SCT, based upon 15 years experience in a large unit performing both adult and paediatric transplants. PMID- 12639527 TI - Evaluation of recurrent gastric malignancy with [F-18]-FDG positron emission tomography. AB - AIM: We retrospectively assessed the use of [(18)F] fluorodeoxyglucose positron emission tomography (FDG PET) in the evaluation of recurrent disease in patients with history of gastric malignancy. MATERIALS AND METHODS: Eighteen patients were referred for FDG PET for evaluation of recurrent gastric cancer. Prior treatments included total (n = 4) or partial gastrectomy (n = 14) followed by chemotherapy alone (n = 7) or combined chemoradiation therapy (n = 2). The interval between the most recent treatment and PET ranged from 3 months to 2 years. Correlative diagnostic data were available in 16 patients and were all obtained within 3 months of the PET study. Validation was by clinical or imaging follow-up (2-45 months) in 16 patients and histology in two patients. RESULTS: PET was concordant with computed tomography (CT) in 12 patients (5 TP, 6 TN, 1 FN). In one patient with negative imaging studies, an incidental finding of left obstructive uropathy was determined to be due to metastatic ureteral stricture. Discordant imaging findings were present in four patients (22% of total). PET-detected diffuse metastatic lesions in three of these patients with rising serum tumour markers while other imaging studies were negative. Additional chemotherapy was initiated in these three patients (17% of total) based on PET localization of disease. PET and a gastric anastomosis biopsy were negative in another patient with positive CT. The remaining two patients without correlative imaging studies died shortly after positive PET studies with presumed recurrent cancer. CONCLUSION: FDG PET may be useful in the evaluation of recurrent gastric cancer, and can localize the disease when CT is non-diagnostic. Imaging evaluation with PET may also impact on the clinical management of patients with recurrent gastric cancer. PMID- 12639528 TI - Percutaneous laser photocoagulation of osteoid osteomas under CT guidance. AB - AIM: The aim was to evaluate laser photocoagulation and the use of the Bonopty needle system in the treatment of osteoid osteoma. MATERIALS AND METHODS: Five patients with osteoid osteomas were treated with computed tomography (CT)-guided, percutaneous laser photocoagulation using the Bonopty biopsy system. RESULTS: Complete pain relief was obtained in four patients. In one patient, pain persisted until the 6 weeks follow-up but resolved within 24 h of repeating the procedure. There were no complications, and patients remained symptom free at follow-up of 4-23 months (mean, 14 months). CONCLUSION: CT-guided laser photocoagulation of osteoid osteoma is a minimally invasive technique that represents a cost-effective alternative to surgical excision. The Bonopty needle system allows successful penetration of the sclerotic bone surrounding the nidus with manual pressure alone. PMID- 12639529 TI - Evaluation of spinal ultrasound in spinal dysraphism. AB - AIMS: The aims of this study were to evaluate the role of spinal ultrasound in detecting occult spinal dysraphism (OSD) in neonates and infants, and to determine the degree of agreement between ultrasound and magnetic resonance imaging (MRI) findings. MATERIALS AND METHODS: Eighty-five consecutive infants had spinal ultrasound over 31 months. Of these, 15 patients (age 1 day-7 months, mean 40 days; nine male) had follow-up MRI. Ultrasound and MRI findings were correlated retrospectively. RESULTS: Six out of 15 (40%) ultrasound examinations showed full agreement with MRI, seven of 15 (47%) had partial agreement, and two of 15 (13%) had no agreement. In the present series ultrasound failed to visualize: four of four dorsal dermal sinuses, three of four fatty filum terminales, one of one terminal lipoma, two of four partial sacral agenesis, three of four hydromyelia and one of 10 low-lying cords. CONCLUSION: Agreement between ultrasound and MRI was good, particularly for the detection of low-lying cord (90%). Therefore we recommend ultrasound as a first-line screening test for OSD. If ultrasound is abnormal, equivocal or technically limited, MRI is advised for full assessment. PMID- 12639530 TI - Audit of a policy of magnetic resonance imaging with diffusion-weighted imaging as first-line neuroimaging for in-patients with clinically suspected acute stroke. AB - AIM: To audit the feasibility and use of diffusion-weighted (DW) magnetic resonance imaging (MRI) as initial neuroimaging for in-patients with clinically suspected acute stroke. MATERIALS AND METHODS: In April 2000, MRI with DW and T2 weighted sequence was locally instituted as initial neuroimaging for patients with clinically suspected acute stroke. This retrospective study reviewed imaging performed for in-patients with suspected acute stroke over a 9-month period. Data were collected on image type, result and need for repeat imaging. RESULTS: During the study period, 124 patients had neuroimaging for suspected cerebrovascular accident, and 119 were MRI safe. Eighty-eight (73.9%) patients underwent DW MRI as first-line investigation. Five patients were not MRI safe and 31 had computed tomography (CT) as first-line imaging due to lack of available MRI capacity. Repeat neuroimaging was performed in 16 (12.9%) patients. Study times were comparable for both types of neuroimaging: a mean of 13 min for MRI and 11 min for CT. CONCLUSION: The audit standard was achieved in 88 (73.9%) patients. The use of DW MRI as a first-line investigation for patients with a clinical diagnosis of acute stroke is achievable in a district general hospital setting. PMID- 12639531 TI - Confidence intervals and sample size calculations for studies of film-reading performance. AB - The relaxation of restrictions on the type of professions that can report films has resulted in radiographers and other healthcare professionals becoming increasingly involved in image interpretation in areas such as mammography, ultrasound and plain-film radiography. Little attention, however, has been given to sample size determinations concerning film-reading performance characteristics such as sensitivity, specificity and accuracy. Illustrated with hypothetical examples, this paper begins by considering standard errors and confidence intervals for performance characteristics and then discusses methods for determining sample size for studies of film-reading performance. Used appropriately, these approaches should result in studies that produce estimates of film-reading performance with adequate precision and enable investigators to optimize the sample size in their studies for the question they seek to answer. PMID- 12639532 TI - The antenatal diagnosis of migration disorders: a series of four cases. PMID- 12639533 TI - De novo formation of a cavernous malformation of the brain in the presence of a developmental venous anomaly. PMID- 12639534 TI - Optimal length of ureteric stents. PMID- 12639535 TI - Magnetic resonance angiography for the diagnosis of renal artery stenosis. PMID- 12639538 TI - Pyridazinones as selective cyclooxygenase-2 inhibitors. AB - Pyridazinone was found to be an excellent core template for selective COX-2 inhibitors. Two potent, selective and orally active COX-2 inhibitors, which were highly efficacious in rat paw edema and rat pyresis models, have been obtained. PMID- 12639540 TI - Synthesis, antiviral activity and pharmacokinetics of P1/P1' substituted 3 aminoindazole cyclic urea HIV protease inhibitors. AB - A series of P1/P1' substituted cyclic urea analogues were prepared in an attempt to increase the intra-cellular antiviral potency of the nonsymmetrical 3 aminoindazoles DMP 850 and DMP 851. The effect of alkyl substitution of the P1/P1' residues on cellular antiviral potency, protein binding, resistance profile and pharmacokinetics are described. PMID- 12639539 TI - Synthesis of N,N',N"-trisubstituted thiourea derivatives and their antagonist effect on the vanilloid receptor. AB - Twenty-seven N,N',N"-trisubstituted thiourea derivatives were prepared. Among them, 1-[3-(4'-hydroxy-3'-methoxy-phenyl)-propyl]-1,3-diphenethyl-thiourea (8l, IC(50)=0.32 microM), showed 2-fold higher antagonistic activity than that of capsazepine (3, IC(50)=0.65 microM) against the vanilloid receptor in a (45)Ca(2+)-influx assay. PMID- 12639542 TI - Synthesis of non-natural C2-homo-ceramide and its apoptotic activity against HL 60 cells. AB - Non-natural ceramide analogues, C2-homo-ceramide and C2-homo-dihydroceramide, were prepared from L-aspartic acid via L-homo-serine. The apoptotic activities of the synthesized ceramide analogues were examined in HL-60 human leukemia cells. C2-homo- and C2-bishomo-ceramide indicate low but considerable apoptotic activities in comparison with C2-ceramide. PMID- 12639541 TI - Synthesis of 6-formyl-pyridine-2-carboxylate derivatives and their telomerase inhibitory activities. AB - Twenty-one pyridine-2-carboxylate derivatives were prepared by the coupling of 6 formyl-2-carboxylic acid with the corresponding phenol, thiophenol, and aniline, substituted with various functional groups. Among them, the 3,4 dichlorothiophenol ester (9p) showed the highest in vitro telomerase inhibitory activity and quite significant in vivo tumor suppression activity. PMID- 12639543 TI - Determination of active components in rhubarb and study of their hydrophobicity by micellar electrokinetic chromatography. AB - A micellar electrokinetic chromatographic (MEKC) method has been developed for the determination of five anthraquinones and one distyrene derivative in rhubarb. The separation conditions were optimized and two kinds of rhubarb plants and rhubarb-containing medicines were analyzed. The negatively charged solutes migrated toward the anode and were retarded by their interaction with the micelle. Hydrophobicity of the solutes was studied by both MEKC with SDS and SDS free capillary zone electrophoresis in the buffer of 15 mmol/L NaH(2)PO(4)+ 20 mmol/L borax and 15% ethanol (v/v). Linear correlation between log k' and log P(OW) was obtained for the five anthraquinones in SDS micelle system. The capacity factor, k', and free energy differences delta(deltaG) derived from this method provided fundamental information on the interaction between the solutes and the micelle. PMID- 12639544 TI - Enterolosaponins A and B, novel triterpene bisdesmosides from Enterolobium contortisiliquum, and evaluation for their macrophage-oriented cytotoxic activity. AB - Two novel triterpene bisdesmosides, designated as enterolosaponin A (1) and B (2), were isolated from Enterolobium contortisiliquum. The chemical structures of 1 and 2 were determined by analysis of their extensive spectroscopic data, as well as hydrolysis followed by chromatographic study. Enterolosaponins have a 2 amino-2-deoxy-D-glucosyl unit (D-glucosamine) as one of the monosaccharides constituting their oligosaccharide moieties, which have been rarely found in natural product research. Enterolosaponin A (1) exhibited a highly selective cytotoxicity against BAC1.2F5 mouse macrophages, and it should be notable that the macrophage death caused by 1 was shown to be neither necrotic nor due to induction of apoptosis from morphology of the died cells, whose cytosol occurred in vacuolation. PMID- 12639545 TI - Synthesis of dopamine transporter selective 3-[2-(diarylmethoxyethylidene)]-8 alkylaryl-8-azabicyclo[3.2.1]octanes. AB - A series of 3-[2-(diarylmethoxyethylidene)]-8-alkylaryl-8 azabicyclo[3.2.1]octanes was synthesized and the binding affinities of the compounds were determined at the dopamine and serotonin transporters. The 8 phenylpropyl analogues 8a (K(i)=4.1 nM) and 8b (K(i)=3.7 nM) were the most potent compounds of the series with binding affinities 3 times greater than GBR-12909. In addition, 8a (SERT/DAT=327) was over 300-fold more selective for the dopamine transporter than the serotonin transporter. PMID- 12639546 TI - Identification of a high-affinity phosphopeptide inhibitor of Stat3. AB - Stat3 is a latent transcription factor that exhibits elevated activity in a variety of human cancers. To find a lead peptide for peptidomimetic drug development we synthesized and tested phosphopeptides derived from known receptor docking sites and found Y(p)LPQTV as the optimal sequence. SAR studies showed that each residue from pY to pY+3 provided binding energy. PMID- 12639547 TI - Discovery and biological evaluation of potent dual ErbB-2/EGFR tyrosine kinase inhibitors: 6-thiazolylquinazolines. AB - We have identified a novel class of 6-thiazolylquinazolines as potent and selective inhibitors of both ErbB-2 and EGFR tyrosine kinase activity, with IC(50) values in the nanomolar range. These compounds inhibited the growth of both EGFR (HN5) and ErbB-2 (BT474) over-expressing human tumor cell lines in vitro. Using xenograft models of the same cell lines, we found that the compounds given orally inhibited in vivo tumor growth significantly compared with control animals. PMID- 12639548 TI - Synthesis and preliminary biological evaluation of 6-O-[11C] [(methoxymethyl)benzyl]guanines, new potential PET breast cancer imaging agents for the DNA repair protein AGT. AB - Novel radiolabeled O(6)-benzylguanine derivatives, 6-O-[(11)C] [(methoxymethyl)benzyl]guanines ([(11)C]p-O(6)-MMBG, 1a; [(11)C]m-O(6)-MMBG, 1b; ([(11)C]o-O(6)-MMBG, 1c), have been synthesized for evaluation as new potential positron emission tomography (PET) breast cancer imaging agents for DNA repair protein, O(6)-alkylguanine-DNA alkyltransferase (AGT). PMID- 12639549 TI - Inhibition of inosine monophosphate dehydrogenase (IMPDH) by 2-[2-(Z) fluorovinyl]inosine 5'-monophosphate. AB - Inosine 5'-monophosphate dehydrogenase (IMPDH; EC 1.1.1.205) isolated from Escherichia coli B3 cells was strongly inhibited by 2-[2-(Z)-fluorovinyl]inosine 5'-monophosphate (2-FVIMP). Inhibition of IMPDH appears to be irreversible with k(inact) and K(i) values of 0.0269 s(-1) and 1.11 microM, respectively. PMID- 12639550 TI - Structure-activity relationship of linear peptide Bu-His6-DPhe7-Arg8-Trp9-Gly10 NH2 at the human melanocortin-1 and -4 receptors: DPhe7 and Trp9 substitution. AB - A series of pentapeptides, based on hMC4R pentapeptide agonist (Bu-His(6)-DPhe(7) Arg(8)-Trp(9)-Gly(10)-NH(2)), was prepared in which either DPhe(7) or Trp(9) residue was systematically substituted. A number of interesting DPhe surrogates (D-Thi, D-3-CF(3)Phe, D-2-Nal and D-3,4-diClPhe) as well as Trp surrogates (2-Nal and Bta) were identified in this study. PMID- 12639551 TI - Protective effect of imidazolopyrazinone antioxidants on ischemia/reperfusion injury. AB - A series of 2-substituted 3,7-dihydroimidazolo[1,2-a]pyrazine-3-ones has been synthesized and evaluated for their antioxidant activity. Compounds 1-8 are inhibitors of AAPH-induced lipid peroxidation (in vitro) and excellent protectors against microvascular damages in ischemia/reperfusion (in vivo). Hence, the bicyclic structure typical of coelenterazine (luciferin) could be considered as a useful lead in medicinal chemistry. PMID- 12639552 TI - Design, synthesis and biological evaluation of benzimidazole/benzothiazole and benzoxazole derivatives as cyclooxygenase inhibitors. AB - We have synthesised a series of 2-[[2-alkoxy-6-pentadecylphenyl)methyl]thio]-1H benzimidazoles/benzothiazoles and benzoxazoles from anacardic acid and investigated their ability to inhibit human cyclooxygenase-2 enzyme (COX-2). The active compounds were screened for cyclooxygenase-1 (COX-1) inhibition. Compound 13 is 384-fold and 19 is more than 470-fold selective towards COX-2 compared to COX-1. Thus, this class of compounds may serve as excellent candidates for selective COX-2 inhibition. PMID- 12639553 TI - Synthesis of fluorescence-labeled sphingosine and sphingosine 1-phosphate; effective tools for sphingosine and sphingosine 1-phosphate behavior. AB - A fluorescence-labeled sphingosine and sphingosine 1-phosphate have been successfully synthesized from the oxazolidinone methyl ester derived from glycidol via monoalkylation and the stereoselective reduction of the resulting ketone. The labeled sphingosine was converted into its phosphate by treatment with sphingosine kinase 1 (SPHK1) from mouse, and in platelets, and it was incorporated into the Chinese Hamster Ovarian (CHO) cells. In addition, MAPK was activated by NBD-Sph-1-P through Edg-1, Sph-1-P receptor. PMID- 12639554 TI - Optimisation of aryl substitution leading to potent methionyl tRNA synthetase inhibitors with excellent gram-positive antibacterial activity. AB - Optimisation of the left-hand-side aryl moiety of a file compound screening hit against Staphylococcus aureus methionyl tRNA synthetase led to the identification of a series of potent nanomolar inhibitors. The best compounds showed excellent antibacterial activity against staphylococcal and enterococcal pathogens, including strains resistant to clinical antibiotics. PMID- 12639555 TI - Synthesis and anti-influenza evaluation of orally active bicyclic ether derivatives related to zanamivir. AB - We synthesized bicyclic ether sialidase inhibitors such as tetrahydro-furan-2-yl, tetrahydro-pyran-2-yl, and oxepan-2-yl derivatives related to zanamivir. These compounds substituted by diol at the C-3' and C-4' positions resulted in the retention of low nanomolar inhibitory activities against not only influenza A virus sialidase but also influenza A virus in cell culture. Compound 11a in particular showed comparable efficacy in vivo relative to that of oseltamivir phosphate. PMID- 12639556 TI - Ganglioside binding pattern of CD33-related siglecs. AB - Our study deals with the interaction of CD33 related-siglecs-5,-7,-8,-9,-10 with gangliosides GT1b, GQ1b, GD3, GM2, GM3 and GD1a. Siglec-5 bound preferentially to GQ1b, but weakly to GT1b, whereas siglec-10 interacted only with GT1b ganglioside. Siglec-7 and siglec-9 displayed binding to gangliosides GD3, GQ1b and GT1b bearing a disialoside motif, though siglec-7 was more potent; besides, siglec-9 interacted also with GM3. Siglec-8 demonstrated low affinity to the gangliosides tested compared with other siglecs. Despite high structural similarity of CD33 related siglecs, they demonstrated different ganglioside selectivity, in particular to the Neu5Acalpha2-8Neu5Ac motif. PMID- 12639557 TI - Exploring new non-sugar sulfated molecules as activators of antithrombin. AB - New non-sugar, small, sulfated molecules, based on our de novo rationally designed activator (-)-epicatechin sulfate (ECS), were investigated to bind and activate antithrombin, an inhibitor of plasma coagulation enzyme factor Xa. For the activators studied, the equilibrium dissociation constant (K(D)) of the interaction with plasma antithrombin varies nearly 53-fold, with the highest affinity of 1.8 microM observed for morin sulfate, while the acceleration in factor Xa inhibition varies 2.6-fold. The results demonstrate that antithrombin binding and activation is a common property of these small sulfated molecules and suggests plausible directions for designing more potent activators. PMID- 12639558 TI - Antineoplastic agents 390. Isolation and structure of phakellistatin 12 from a Chuuk archipelago marine sponge. AB - A new cancer cell growth inhibitory (P388 lymphocytic leukemia ED(50) 2.8 microg/mL) cyclodecapeptide designated phakellistatin 12 (2) has been isolated as a trace (1.7 x 10(-6)% yield) constituent of the Western Pacific Ocean (Federated States of Micronesia-Chuuk) sponge Phakellia sp. Employing principally a combination of high resolution FAB with high field (500 MHz) 1H, 13C and 2-D NMR and chiral GC analyses the structure (all S chirality) cyclo-Ile-Phe-Thr-Leu-Pro Pro-Tyr-Ile-Pro-Pro was assigned. PMID- 12639559 TI - Synthesis, spectral studies and screening for amoebicidal activity of new palladium(II) complexes derived from thiophene-2-carboxaldehyde thiosemicarbazones. AB - In view of the antiamoebic properties observed for many thiophene-2 carboxaldehyde thiosemicarbazones, a series of N(4)-substituted thiosemicarbazones metal complexes derived from thiophene-2-carboxaldehyde was prepared for evaluation against Entamoeba histolytica. Reaction of thiophene-2 carboxaldehyde with cycloalkylaminothiocarbonylhydrazines having different amines gave the corresponding thiosemicarbazones. Reaction of latter with [Pd(DMSO)(2)Cl(2)] gave requisite palladium thiosemicarbazone complexes of the type [Pd(TSC)Cl(2)] (where TSC=thiosemicarbazones). Screening of antiamoebic activity of these compounds was assayed in vitro against (HM-1:1MSS) strain of E. histolytica. Enhancement of antiamoebic resulted from introducing palladium metal in the thiosemicarbazone moiety. Among the studied compounds, [Pd(2-TCA-1,2,3,4 THQTSC)Cl(2)] (2a) showed better activity. PMID- 12639560 TI - Novel N1-(benzyl)cinnamamidine derived NR2B subtype-selective NMDA receptor antagonists. AB - Novel (E)-N(1)-(benzyl)cinnamamidines were prepared and evaluated as NR2B subtype NMDA receptor ligands. Excellent affinity was achieved by appropriate substitution of either phenyl ring. The 2-methoxybenzyl compound 1h had approximately 1,000-fold lower IC(50) in NR2B than NR2A-containing cells. Replacement of the styryl unit by 2-naphthyl was well tolerated. PMID- 12639561 TI - Orally efficacious NR2B-selective NMDA receptor antagonists. AB - A novel series of benzamidines was synthesized and shown to exhibit NR2B-subtype selective NMDA antagonist activity. Compound 31 is orally active in a carrageenan induced rat hyperalgesia model of pain and shows no motor coordination side effects. PMID- 12639562 TI - Design, synthesis, and discovery of 5-piperazinyl-1,2,6,7-tetrahydro-5H azepino[3,2,1-hi]indol-4-one derivatives: a novel series of mixed dopamine D2/D4 receptor antagonist. AB - 5-piperazinyl-1,2,6,7-tetrahydro-5H-azepino[3,2,1-hi]indol-4-one derivatives were designed, synthesized, and identified as a new series of mixed dopamine D(2)/D(4) receptor antagonists. This series featured a rigid tricyclic ring system as an important pharmacophore core structure for high binding affinity. Molecular modeling studies are also described. PMID- 12639563 TI - Biological evaluation and interconversion studies of rotamers of SCH 351125, an orally bioavailable CCR5 antagonist. AB - The separation and biological evaluation of rotamers as well as interconversion studies on rotamers of our clinical candidate SCH 351125 are described. PMID- 12639564 TI - Oximino-piperidino-piperidine-based CCR5 antagonists. Part 2: synthesis, SAR and biological evaluation of symmetrical heteroaryl carboxamides. AB - The synthesis, SAR and biological evaluation of symmetrical amide analogues of our clinical candidate SCH 351125 are described. A series of potent and orally bioavailable CCR5 antagonists containing symmetrical 2,6-dimethyl isonicotinamides and 2, 6-dimethyl pyrimidines amides were generated with enhanced affinity for the CCR5 receptor. PMID- 12639565 TI - Four novel bis-(naphtho-gamma-pyrones) isolated from Fusarium species as inhibitors of HIV-1 integrase. AB - Integration of viral DNA into host cell DNA is an essential step in retroviral (HIV-1) replication and is catalyzed by HIV-1 integrase. HIV-1 integrase is a novel therapeutic target and is the focus of efforts to identify effective inhibitors that will prevent/or cure HIV infections. Four novel naphtho-gamma pyrones, belonging to the chaetochromin and ustilaginoidin family, were discovered as inhibitors of HIV-1 integrase from the screening of fungal extracts using a recombinant in vitro assay. These compounds inhibit both the coupled and strand transfer activity of HIV-1 integrase with IC(50) values of 1-3 and 4-12 microM, respectively. The discovery, structure elucidation, chemical modification and the structure-activity relationship of these compounds are described. PMID- 12639566 TI - A structure-permeability study of small drug-like molecules. AB - A systematic structure-permeability relationship study on a set of small drug like molecules with log D values in the range -2.5 to 3 and carrying a diverse array of functionality reveals that the compounds with log D>0 and <3 are highly permeable. Surprisingly, several tetrazole derivatives were found to be substrates for efflux pump(s). PMID- 12639567 TI - Design, synthesis, and structure-activity relationships of unsubstituted piperazinone-based transition state factor Xa inhibitors. AB - A series of novel transition state factor Xa inhibitors containing a variety of lactam ring systems as central templates was synthesized in an expedient manner and allowed for a great deal of structural variability. Among them, the piperazinone-based inhibitors were found to be not only active against factor Xa but also selective over thrombin. Optimization of the P4 moiety yielded several potent compounds with IC(50) below 1 nM against factor Xa. PMID- 12639568 TI - Design, synthesis, and structure-activity relationships of substituted piperazinone-based transition state factor Xa inhibitors. AB - The structure-activity relationship of a novel series of substituted piperazinone based factor Xa inhibitors is described. The most potent compound 34 displays IC(50) of 0.9 nM. PMID- 12639569 TI - Binding of nicotine and homoazanicotine analogues at neuronal nicotinic acetylcholinergic (nACh) receptors. AB - A total of 20 substituted analogues of nicotine (1a) and homoazanicotine (3a) were examined in order to determine whether or not they might bind in a similar manner at alpha4beta2 nicotinic acetylcholinergic (nACh) receptors. It was found that parallel structural changes in the two series resulted in parallel shifts in affinity. Evidence suggests that the two series are binding in a comparable fashion. PMID- 12639570 TI - Functional expression and characterization of EryA, the erythritol kinase of Brucella abortus, and enzymatic synthesis of L-erythritol-4-phosphate. AB - The eryA gene of the bacterial pathogen Brucella abortus has been functionally expressed in Escherichia coli. The resultant EryA was shown to catalyze the ATP dependent conversion of erythritol to L-erythritol-4-phosphate (L-E4P). The steady state kinetic parameters of this reaction were determined and the enzyme was used to prepare L-E4P which was shown to be a weak inhibitor of 2-C-methyl-D erythritol-4-phosphate cytidyltransferase (YgbP). PMID- 12639572 TI - Lead discovery of alpha,beta-unsaturated sulfones from a combinatorial library as inhibitors of inducible VCAM-1 expression. AB - alpha,beta-Unsaturated sulfones have been discovered from a combinatorial library as leads for a new series of inhibitors of inducible VCAM-1 expression. Although not essential, further conjugation of the sulfonyl group to another vinyl group or a phenyl group increases the potency dramatically. PMID- 12639571 TI - Substituted aminopyridines as potent and selective phosphodiesterase-4 inhibitors. AB - The synthesis and the biological evaluation of new potent phosphodiesterase type 4 (PDE4) inhibitors are presented. This new series was elaborated by replacement of the metabolically resistant phenyl hexafluorocarbinol of L-791,943 (1) by a substituted aminopyridine residue. The structure-activity relationship of N substitution on 3 led to the identification of (-)-3n which exhibited a good PDE4 inhibitor activity (HWB-TNFalpha=0.12 microM) and an improved pharmacokinetic profile over L-791,943 (rat t(1/2)=2 h). (-)-3n was well tolerated in ferret with an emetic threshold of 30 mg/kg (po) and was found to be active in the ovalbumin induced bronchoconstriction model in guinea pig (54%, 0.1 mg/kg, ip) as well as the ascaris-induced bronchoconstriction model in sheep (64%/97%, early/late, 0.5 mg/kg, iv). PMID- 12639573 TI - 1,4-dibenzylpiperazines possess anticocaine activity. AB - N,N-dibenzylpiperazines have high affinity for sigma receptors, and we aimed to increase their anticocaine activity by introducing substituents known to enhance such activity in other sigma ligands. Ligands with high affinity for sigma-1 receptors resulted, but their activity in attenuating cocaine-induced convulsions did not correlate with sigma-1 binding affinity, and may be more closely related to their sigma-2 binding affinities. PMID- 12639574 TI - Synthesis and biological evaluation of novel indoloazepine derivatives as non peptide vasopressin V2 receptor antagonists. AB - A series of novel 3,4,5,6-tetrahydro-1H-azepino[4,3,2-cd]indoles was synthesized and tested for vasopressin receptor antagonist activity. We identified compounds with high affinity for the human V2 receptor and good selectivity over the human V1a receptor. Compound 6c bound to V2 receptors with an IC(50) value of 20 nM, had >100-fold selectivity over V1a receptors, and inhibited cAMP formation in a cellular V2 functional assay with an IC(50) value of 70 nM. PMID- 12639575 TI - Cryptophycin affinity labels: synthesis and biological activity of a benzophenone analogue of cryptophycin-24. AB - An efficient synthesis of a C16 side chain benzophenone analogue of cryptophycin 24 using a crotylboration reaction and Heck coupling as key steps is described. In an in vitro tubulin assembly assay, the benzophenone analogue of the beta isomer (IC(50)=7.4 microM) is twice as active as cryptophycin-24 (IC(50)=15 microM). PMID- 12639576 TI - Synthesis and biological activities of novel beta-carbolines as PDE5 inhibitors. AB - A series of N(2)-furoyl and N(2)pyrimidinyl beta-carbolines was discovered to possess potent inhibitory activity against PDE5. During the synthesis we developed a tandem resin quenching protocol, which allowed us to synthesize large number of target compounds in a rapid fashion. Representative compounds exhibit superior selectivity to sildenafil versus other isozymes of PDEs, and demonstrated in vivo efficacy in increasing introcavernosal pressure in dogs. PMID- 12639577 TI - Novel, highly potent, selective 5-HT2A/D2 receptor antagonists as potential atypical antipsychotics. AB - The discovery of N-substituted-pyridoindolines and their binding affinities at the 5-HT(2A), 5-HT(2C) and D(2) receptors, and in vivo efficacy as 5-HT(2A) antagonists is described. The structure-activity relationship of a series of core tetracyclic derivatives with varying butyrophenone sidechains is also discussed. This study has led to the identification of potent, orally bioavailable 5 HT(2A)/D(2) receptor dual antagonists as potential atypical antipsychotics. PMID- 12639578 TI - Same-sex attraction in a birth cohort: prevalence and persistence in early adulthood. AB - There is a continuing debate about the importance of social versus biological factors in the expression of same-sex attraction. Investigation of prevalence, continuities, and changes over time among young adults growing up in a country with a relatively accepting climate to homosexuality is likely to illuminate this debate. Analyses were therefore undertaken of self-reported same-sex attraction at age 21 and 26, in a cohort of about 1000 people born in 1972/3 in one New Zealand city. Participants were also asked about same-sex behaviour and attitudes to same-sex relationships. By age 26, 10.7% of men and 24.5% of women reported being attracted to their own sex at some time. This dropped to 5.6% of men and 16.4% of women who reported some current same-sex attraction. Current attraction predominantly to their own sex or equally to both sexes (major attraction) was reported by 1.6% of men and 2.1% of women. Occasional same-sex attraction, but not major attraction, was more common among the most educated. Between age 21 and 26, slightly more men moved away from an exclusive heterosexual attraction (1.9% of all men) than moved towards it (1.0%), while for women, many more moved away (9.5%) than towards (1.3%) exclusive heterosexual attraction. These findings show that much same-sex attraction is not exclusive and is unstable in early adulthood, especially among women. The proportion of women reporting some same sex attraction in New Zealand is high compared both to men, and to women in the UK and US. These observations, along with the variation with education, are consistent with a large role for the social environment in the acknowledgement of same-sex attraction. The smaller group with major same-sex attraction, which changed less over time, and did not differ by education, is consistent with a basic biological dimension to sexual attraction. Overall these findings argue against any single explanation for homosexual attraction. PMID- 12639579 TI - How important is sex in later life? The views of older people. AB - Stereotypes of an asexual old age remain pervasive, shaping not only popular images of older people, but also research and policy agendas. However, older people's own attitudes towards the role and value of sex in later life remain relatively unexplored. This paper draws on both quantitative and qualitative data to examine how sex is prioritised in middle age and later life. Data collection methods involved completion of two quality of life measures (WHOQOL-100 and WHOQOL Importance Scale), followed by semi-structured interviews. In total the sample comprised 69 individuals recruited from the age/sex register of a general practice in Sheffield in the UK. This paper will focus upon the accounts of 21 men and 23 women aged 50-92 years. Ratings of the importance of sex to participants were gathered from the WHOQOL Importance Scale; the in-depth interviews enabled the basis for this prioritisation to be explored. Analysis identified the following key themes. Participants who did not consider sex to be of any importance to them neither had a current sexual partner, nor felt that they would have another sexual partner in their lifetime. Indeed, all participants who had a current sexual partner attributed at least some importance to sex, with many rating sex as 'very' or 'extremely' important. However, experiencing barriers to being sexually active led them to place less importance on sex; this was particularly apparent when health problems and widowhood were experienced. Age was seen as facilitating coping when sex became less frequent, or stopped altogether. This was explained in terms of sexual desire decreasing with age (for some male participants), the cessation of sex being easier to cope with in a relationship of long duration and the expectation that sex will become less possible with 'normal ageing'. The discussion considers the implications of these findings for this developing field. PMID- 12639580 TI - Managing menopause: a qualitative analysis of self-help literature for women at midlife. AB - This article reports on a qualitative social constructionist analysis that aims to examine the ways in which menopause and women's bodies are represented in self help texts. In particular, we aim to compare texts with a more traditional 'medical' approach and others taking a more 'woman-centred' perspective. Four diverse self-help books on menopause and HRT available in England were analysed to examine the ways in which women, menopause and midlife were portrayed, and to investigate the construction of notions of knowledge, expertise and responsibility. The selected texts were published between 1992 and 1996 and covered a range of perspectives, including medical, alternative and feminist. Results showed that menopause was constructed as a 'deficiency disease' in all four texts, although in three of the texts this 'disease' discourse was counterposed by the simultaneous use of a 'menopause as natural' discourse. Menopause was also constructed as inherently complex and confusing, as were women's bodies. A discourse of 'change' was drawn upon in which menopause was portrayed as only one of the stressful events women must cope with at midlife. Finally, most of the texts drew on a discourse of 'management' rather than one of 'treatment' or 'cure' when discussing how menopause, and women's relationship to menopause should be handled. The medical profession was constructed as the primary source of expertise on menopause and women's bodies, although responsibility for the 'management' of menopause as a chronic condition lay solely with individual women. Although there were a number of differences in representations of menopause in medically oriented self-help texts and those adopting a more woman-centred perspective, our analysis also revealed several areas of similarity and overlap with regard to the construction of menopause and its 'management'. The implications of these findings for the construction of menopause in self-help texts for women are discussed. PMID- 12639581 TI - Feminist ethics and menopause: autonomy and decision-making in primary medical care. AB - The construction of menopause as a long-term risk to health and the adoption of discourses of prevention has made necessary a decision by women about medical treatment; specifically regarding the use of hormone replacement therapy. In a study of general practitioners' accounts of menopause and treatment in Australia, women's 'choice', 'informed decision-making' and 'empowerment' were key themes through which primary medical care for women at menopause was presented. These accounts create a position for women defined by the concept of individual choice and an ethic of autonomy. These data are a basis for theorising more generally in this paper. We critically examine the construct of 'informed decision-making' in relation to several approaches to ethics including bioethics and a range of feminist ethics. We identify the intensification of power relations produced by an ethic of autonomy and discuss the ways these considerations inform a feminist ethics of decision-making by women. We argue that an 'ethic of autonomy' and an 'offer of choice' in relation to health care for women at menopause, far from being emancipatory, serves to intensify power relations. The dichotomy of choice, to take or not to take hormone replacement therapy, is required to be a choice and is embedded in relations of power and bioethical discourse that construct meanings about what constitutes decision-making at menopause. The deployment of the principle of autonomy in medical practice limits decision-making by women precisely because it is detached from the construction of meaning and the self and makes invisible the relations of power of which it is a part. PMID- 12639582 TI - Factors predicting choice of treatment for menorrhagia in gynaecology outpatient clinics. AB - The aim of this prospective study was to investigate factors predicting choice of treatment for excessive menstrual bleeding, with special emphasis on women's pre treatment preference. A cohort of women with heavy menstruation and their treatment process in gynaecology outpatient clinics were followed-up for 1yr. A total of 383 35-54-yr-old women attending 14 Finnish hospitals participated. They completed a questionnaire before their first outpatient clinic visit, and postal follow-ups were conducted 3 and 12 months later. Information on treatment(s) during the follow-up was taken from medical records and questionnaires. The choice between hysterectomy and conservative treatments, and fulfillment of pre treatment preference were the main outcome measures. During the 1-yr follow-up, 51% (n=196) of the women underwent hysterectomy and nine were still awaiting it, 12% (n=44) had a minor surgical procedure, 11% (n=41) had oral medication, 9% (n=33) used a hormonal intrauterine system, and nine women changed preventive method. Forty-two women (11%) reported having had no treatment. Data on previous treatments suggested that conservative treatment modalities were under-used. Most of the treatment decisions were made within the first 3-month period. Women's pre treatment preference was the strongest predictor of chosen treatment. Unemployment, irregular periods and anxiety decreased the probability of a decision for hysterectomy, while pelvic pain and inconvenience due to bleeding increased it. The treatment plan accorded with pre-treatment preference in 72% of the women preferring hysterectomy and in 74% of those preferring a conservative option. PMID- 12639583 TI - Elderly people's ratings of the importance of health-related factors to their self-assessments of health. AB - Identifying the bases for self-assessed health (SAH) has interested researchers in their attempts to understand its validity as a predictor of future health outcomes. Quantitative approaches typically used statistical methods to identify correlates of SAH while qualitative approaches asked people to elaborate on the reasons underlying their rating of health. The current study used a quantitative methodology, asking 487 elderly people to rate the importance of 42 health related factors as bases for their SAH judgment. Factors indicating overall functioning/vitality were rated highly by all participants. Factors indicating current disease were rated highly by people reporting poor/fair SAH while risk factors and positive indicators were rated highly by those reporting good, very good, or excellent health. Thus, there seems to be a clear distinction between poor and fair SAH that reflect levels of illness, and higher levels of SAH that reflect levels of health. PMID- 12639584 TI - What lies behind the subjective evaluation of health status? AB - The evaluation of health or subjective health (SH) is considered a legitimate indicator of overall health status, providing a valid, reliable and cost effective means of health assessment. This study looks at factors reported by respondents as influencing their SH, it analyzes to which extent people compare themselves with others when evaluating their health, and examines the difference between people with optimal or sub-optimal SH. Face-to-face interviews were conducted with 383 Israeli residents. Three models for judging health status were identified: the biomedical or disease oriented, the emotional or "general feeling", and the functional-related model. The reported influence of some factors for the evaluation of health changed by age and by level of subjective health. Respondents with sub-optimal health reported tiredness and pain as more influential. Most interviewees spontaneously reported comparing their health to reference groups. Age and level of subjective health were associated with the way people compare their health to others. The young reporting sub-optimal health did not compare themselves to people their age, but a high percentage of the old did so. Among those with excellent health, the young rather than the old were more likely to compare themselves to people their age. These findings imply that each individual tries to find ways to evaluate his/her health in a more positive light. When old and not healthy they tend to compare themselves to friends or people their age, but if young and not healthy they do not perform the comparison so as not to feel worse. Understanding how people evaluate their health can contribute to the conceptual development of subjective health. PMID- 12639585 TI - Undernutrition in Benin-an analysis based on graphical models. AB - Undernutrition is one of the most important health problems in developing countries. Examining its determinants implies the investigation of a complex association structure including a large number of potential influence variables and different types of influences. A recently developed statistical technique to cope with such situations are graphical chain models. In this paper, this approach is used to investigate the determinants of undernutrition in Benin (West Africa). Since this method also reveals indirect influences, interesting insight is gained into the association structure of all variables incorporated. The analysis identifies mother's education, socioeconomic status, and religion as three variables with particularly strong direct and indirect linkages to undernutrition. PMID- 12639586 TI - Developing a framework for assessment of the environmental determinants of walking and cycling. AB - The focus for interventions and research on physical activity has moved away from vigorous activity to moderate-intensity activities, such as walking. In addition, a social ecological approach to physical activity research and practice is recommended. This approach considers the influence of the environment and policies on physical activity. Although there is limited empirical published evidence related to the features of the physical environment that influence physical activity, urban planning and transport agencies have developed policies and strategies that have the potential to influence whether people walk or cycle in their neighbourhood. This paper presents the development of a framework of the potential environmental influences on walking and cycling based on published evidence and policy literature, interviews with experts and a Delphi study. The framework includes four features: functional, safety, aesthetic and destination; as well as the hypothesised factors that contribute to each of these features of the environment. In addition, the Delphi experts determined the perceived relative importance of these factors. Based on these factors, a data collection tool will be developed and the frameworks will be tested through the collection of environmental information on neighbourhoods, where data on the walking and cycling patterns have been collected previously. Identifying the environmental factors that influence walking and cycling will allow the inclusion of a public health perspective as well as those of urban planning and transport in the design of built environments. PMID- 12639587 TI - Social and health determinants of the efficiency of cotton farmers in Northern Cote d'Ivoire. AB - This article assesses the role of malaria and some social determinants on the agricultural development and more precisely on efficiency in the context of cotton crop in the Korhogo region in the North of Cote d'Ivoire. Data envelopment analyses (DEA) was first applied for the purpose of calculating relative efficiencies in production. A Tobit regression model was then used to explain the variation in the DEA scores and check the hypotheses that the efficiency deviations between farmers can be explained by the disparity of malaria morbidity rate among the farmers and their family, by social cohesiveness and cultural behaviour. Field data were collected by the authors between March 1997 and February 1998 on 700 rural households living in three rice production systems differently exposed to the malaria risk. Two malaria indicators were used for the active (11-55 years old) family members of the farm: Plasmodium falciparum infection rate and high parasite density infection rate. The DEA model was applied on the sub-sample of cotton growers (about one third of the households of the full sample). Results of the different DEA and Tobit models (depending of the production process hypothesis) show that high parasite density infection has a direct and indirect negative effect on efficiency in the cotton crop. They also show that more cotton growers in the village improve efficiency, although villages where cotton is growing more widespread have weaker social cohesion. PMID- 12639588 TI - Indigenous social insurance as an alternative financing mechanism for health care in Ethiopia (the case of eders). AB - With increasing demand for services further propagated by population growth and by people's response to newly emerging pathologies, nations in sub-Saharan Africa are faced with insurmountable problems in sustaining their health systems. Realizing the inadequacy of solely relying on the public sector, these countries are seeking alternative mechanisms for health financing. Among the alternatives suggested are risk-sharing mechanisms that include community-based schemes that tap the potential of indigenous social arrangements. In Ethiopia, eders are major forms of indigenous arrangements utilized mainly for assisting victims in bereavement and executing funeral-related activities. These associations are also called upon in various self-help activities and sometimes provide health insurance, even though mostly in an informal manner. Therefore, they have the potential to serve as social financing mechanisms. Since these are already functioning groups, the administrative cost for the extra health-related activity will not be as high as in the case of forming a new insurance entity. In addition, the fact that eders are based on mutual understanding among members minimizes the possibility of adverse selection. Based on the above background, an exploratory study was conducted in 40 villages distributed in various parts of Ethiopia to assess the possible roles eders might play in providing insurance for health financing. Both qualitative and quantitative (household and health facility exit interview surveys) methods of data collection were utilized. The study concludes that eder-based schemes are, indeed, options for experimentation as mechanisms for financing health care in rural Ethiopia. It was also found that 21.5% of respondents in the household and 16% of those in the exit surveys were already utilizing eders to finance part of their health expenditure. In addition, 86% of the respondents in the household and 90% of those in the exit survey were willing to participate in eder-based health insurance schemes. PMID- 12639590 TI - Self-esteem and women with disabilities. AB - This study examines the sense of self of women with physical disabilities in terms of self-esteem, self-cognition (perceptions of how others see them), and social isolation. It was hypothesized that these variables mediate the relation of precursor variables (age, education, severity of disability, and childhood experiences, including overprotection, familial affection, and school environment) and outcomes (intimacy, employment, and health promoting behaviors). Data were gathered from a sample of 881 community-dwelling women in the USA, 475 with a variety of mild to severe physical disabilities, and 406 without disabilities. Correlation analyses indicated that the women with disabilities had significantly lower self-cognition and self-esteem, and greater social isolation than the women without disabilities, as well as significantly less education, more overprotection during childhood, poorer quality of intimate relationships, and lower rates of salaried employment. Path analysis indicated that each of the sense of self mediators was significantly related to the outcome of intimacy, that both social isolation and self-esteem were significantly related to health promoting behaviors, and that only self-esteem was significantly related to employment. Respondents who were older, less disabled, less educated, less over protected, and had more affection shown in the home tended to feel that others saw them more positively. Women with positive school environments, less over protection, and more affection in the home experienced less social isolation; age, education, and disability severity were not significantly related to social isolation. Older respondents with less disability, a more positive school environment, less over-protection, and more affection in the home tended to have greater self-esteem; education was not significantly related to self-esteem. Older respondents tended to report less intimacy. Younger, more educated, and less disabled respondents were significantly more likely to be employed. More highly educated respondents reported engaging in more health promoting behaviors. PMID- 12639589 TI - The influence of cost-effectiveness information on physicians' cancer screening recommendations. AB - Physicians are increasingly faced with choices in which one screening strategy is both more effective and more expensive than another. One way to make such choices is to examine the cost-effectiveness of the more costly strategy over the less costly one. However, little is known about how cost-effectiveness information influences physicians' screening decisions. We surveyed 900 primary care US physicians, and presented each with a hypothetical cancer-screening scenario. We created three familiar screening scenarios, involving cervical, colon, and breast cancer. We also created three unfamiliar screening scenarios. Physicians were randomized to receive one of nine questionnaires, each containing one screening scenario. Three questionnaires posed one of the familiar screening scenarios without cost-effectiveness information, three posed one of the familiar scenarios with cost-effectiveness information, and three posed one of the unfamiliar scenarios with cost-effectiveness information. The cost-effectiveness information for familiar scenarios was drawn from the medical literature. The cost effectiveness information for unfamiliar scenarios was fabricated to match that of a corresponding familiar scenario. In all questionnaires, physicians were asked what screening alternative they would recommend. A total of 560 physicians responded (65%). For familiar scenarios, providing cost-effectiveness information had at most a small influence on physicians' screening recommendations; it reduced the proportion of physicians recommending annual Pap smears (p=0.003), but did not significantly alter the aggressiveness of colon cancer and breast cancer screening (both p's<0.1). For all three unfamiliar scenarios, physicians were significantly less likely to recommend expensive screening strategies than in corresponding familiar scenarios (all p's<0.001). Physicians' written explanations revealed a number of factors that moderated the influence of cost effectiveness information on their screening recommendations. Providing physicians with cost-effectiveness information had only a moderate influence on their screening recommendations for cervical, colon, and breast cancer. Significantly, fewer physicians recommended aggressive screening for unfamiliar cancers than for familiar ones, despite similar cost-effectiveness. Physicians are relatively reluctant to abandon common screening strategies, even when they learn that they are expensive, and are hesitant to adopt unfamiliar screening strategies, even when they learn that they are inexpensive. PMID- 12639591 TI - "Good jobs" to "bad jobs": replicated evidence of an employment continuum from two large surveys. AB - The goal of this study was to offer an expanded conceptualization of the employment continuum and test its utility by examining the association of different employment statuses with physical health and depression. Using data from two large cross-sectional surveys we develop five different employment categories (i.e., "optimal", "economically good", "psychologically good", "barely adequate", and "inadequate" employment) in addition to unemployment to form an employment continuum. Evidence from these studies indicates that less than optimal forms of employment are not randomly distributed throughout the population, and that a substantial number of adults are working in "inadequate or "barely adequate" jobs. Moreover, our analyses revealed a consistent association between less than optimal jobs and poorer physical and mental health among adults. We conclude our paper by discussing important policy implications of these findings and offering suggestions for future research. PMID- 12639592 TI - Quality of life in an urban Asian population: the impact of ethnicity and socio economic status. AB - The relationships between ethnicity, socio-economic status (SES) and health related quality of life (HRQoL) have not been well characterised in most Asian populations. We therefore studied the influence of ethnicity and SES on HRQoL in a multi-ethnic urban Asian population, adjusting for the influence of other known determinants of HRQoL. In a disproportionately stratified, cross-sectional, population-based survey, Chinese, Malay and Indian subjects in Singapore completed the Short Form 36 Health Survey (SF-36) HRQoL measure and were assessed to determine demographic, socio-economic, psychosocial and other characteristics. Multiple linear regression models were used to study the influence of ethnicity and SES on SF-36 scores while adjusting for the influence of other determinants of HRQoL. The survey participation rate was 92.8%. Ethnic differences in HRQoL were present for all 8 SF-36 scales (p<0.001 for all scales except General Health) among the 4122 Chinese, Malays and Indians surveyed. These ethnic groups also differed in several known determinants of HRQoL (e.g., Chinese had more years of education and Indians had more chronic medical conditions). After adjusting for the influence of these factors, ethnicity and SES independently influenced HRQoL, with mean differences in SF-36 scores due to ethnicity ranging from 1.4 to 13.1 points. Educational level and housing type (markers of SES) were also associated with SF-36 scores (0.5-0.6 point increase per year of education and 3.5-4.0 point increase with better housing type, respectively). Better HRQoL was also associated with better family support, and poorer HRQoL with acute and chronic medical conditions and sick days. The study concludes that ethnicity and SES are associated with clinically important differences in HRQoL in a multi ethnic, urban Asian population. PMID- 12639593 TI - Primary health and social care services in the UK: progress towards partnership? AB - There has been a shift in the theoretical debates around the ways in which organisations deliver the state's objectives of providing health and social care services for its citizens, focusing on issues of welfare governance and the encouragement of partnership working between organisations. This article develops these theories by focusing on developments in primary health and social care policy in the UK, which have undergone a radical change recently. Responsibility for commissioning health care services now falls to primary care groups and trusts (PCG/Ts), run by general practitioners, other primary care practitioners, managers and lay members, and there is some pressure on primary care groups and primary care trusts to engage in partnership working with social services, for example, to cut hospital waiting lists or provide intermediate care services. One policy option is for these organisations to form Care Trusts, integrating the commissioning of health and social care for older people and ending the historical organisational divisions between health and social care in the UK. This paper examines evidence from the first stage of a 3-year longitudinal quantitative and qualitative study of the development of partnership working between PCG/Ts and social services departments in England. It examines whether the evidence suggests that the integration of health and social care is feasible or desirable in older people's services. PMID- 12639594 TI - Suicide attempts in the United States: the role of physical illness. AB - The study aimed to determine the relationship between physical illness, mental disorder, and the likelihood of suicide attempt among adults aged 15-54 in the United States. Data were drawn from the National Comorbidity Survey (N=8,098), a national probability sample of adults in the United States. Multivariate logistic regression analyses were used to determine the relationship between self-reported physical illness and the likelihood of suicide attempt. Lung disease (OR=1.8 (1.1, 2.7)), ulcer (OR=2.1 (1.3, 3.4)), and AIDS (OR=44.1 (10.5, 185.6)) were each associated with a significantly increased likelihood of suicide attempt, independent of the effects of mental disorders. Consistent with previous studies, the number of physical illnesses was linearly related to an increased odds of suicide attempt (OR=1.3 (1.2, 1.5)). Possible mechanisms for these associations are discussed. These findings call for the inclusion of a range of physical health problems, especially chronic illnesses, in future research on suicide attempts in the population. PMID- 12639595 TI - Predictors of adherence to treatment for latent tuberculosis infection in high risk Latino adolescents: a behavioral epidemiological analysis. AB - The objective was to test whether theoretical variables predict adherence to treatment for latent tuberculosis infection in high-risk Latino adolescents. 286 Latino adolescents, age 13-18 years, were recruited from 10 middle/high schools in San Diego County, San Diego, USA. Participants completed a baseline interview and up to 9 monthly interviews. The cumulative number of pills consumed in 9 months was regressed on 16 independent variables, entered hierarchically in seven blocks. The final model accounted for 25% of the variance in adherence to isoniazid (INH), F (16, 230)=4.69, p<0.001. Adherence counseling (+), age (-), grades (+), being bicultural (+), and risk behaviors (-) were significantly related to adherence. Learning theories presume that adherence to medical regimens requires social support and freedom from physical and social barriers. Results support these theories. Future studies should explore additional precepts in order to identify additional predictors and to maximize adherence to INH among Latino adolescents and other high-risk populations. Doing so should decrease the risk of active TB among high-risk racial/ethnic and foreign-born populations. PMID- 12639596 TI - Social capital and neighborhood mortality rates in Chicago. AB - Several empirical studies have suggested that neighborhood characteristics influence health, with most studies having focused on neighborhood deprivation or aspects of the physical environment, such as services and amenities. However, such physical characteristics are not the only features of neighborhoods that potentially affect health. Neighborhoods also matter because of the nature of their social organization. This study examined social capital as a potential neighborhood characteristic influencing health. Using a cross-sectional study design which linked counts of death for persons 45-64 years by race and sex to neighborhood indicators of social capital and poverty for 342 Chicago neighborhoods in the USA, we tested the ecological association between neighborhood-level social capital and mortality rates, taking advantage of the community survey data collected as part of the Project on Human Development in Chicago Neighborhoods. We estimated a hierarchical generalized linear model to examine the association of race and sex specific mortality rates to social capital. Overall, neighborhood social capital-as measured by reciprocity, trust, and civic participation-was associated with lower neighborhood death rates, after adjustment for neighborhood material deprivation. Specifically, higher levels of neighborhood social capital were associated with lower neighborhood death rates for total mortality as well as death from heart disease and "other" causes for White men and women and, to a less consistent extent, for Blacks. However, there was no association between social capital and cancer mortality. Although, the findings from this study extend the state-level findings linking social capital to health to the level of neighborhoods, much work remains to be carried out before social capital can be widely applied to improve population health, including establishing standards of measurement, and exploring the potential "downsides" of social capital. PMID- 12639597 TI - Economy, community and mortality in British Columbia, Canada. AB - Stimulated by the growing body of literature relating economic inequalities to inequalities in health, this article explores relationships between various economic attributes of communities and mortality rates among 24 coastal communities in British Columbia, Canada. Average household income, a measure of community wealth, was negatively related and the incidence of low incomes, a measure of poverty, was positively related to age-standardized mortality. Both were more strongly related to female than male mortality. Mean and median household income, the incidence of low incomes and a lack of disposable income, and the proportion of total income dollars derived from government sources were significantly related to mortality rates for younger and middle-aged men but not for elderly men. Mortality rates for younger and middle-aged women were not explicated by these economic attributes of communities: among elderly women only, mortality rates were higher in communities with a lower average household income and in those with a higher incidence of low incomes. Finally, a higher concentration in white-collar industries was related to higher mortality rates for females, even after controlling for other economic attributes of communities. These results do not obviously support a psychosocial argument for an individual level relationship between income and health that assumes residents perceive their status primarily in relation to other members of the same community, but do provide moderate support for the materialist argument and moderate support for the psychosocial argument that assumes community residents perceive their status in relation to an encompassing reference group. Other viable interpretations of these relationships pertain to ecological characteristics of communities that are related to both economic well-being and population health status; in this instance, concentration in specific economic industries may help to understand the ecological relationships presented here. PMID- 12639598 TI - Social relations and self-reported health: a prospective analysis of the French Gazel cohort. AB - Social networks and social support are strongly associated with health, yet the pathways between social relations and health are not well understood. In one of the very few studies on this issue conducted in France, we used data from the French GAZEL cohort of employees of the national gas and electricity company, to (1) explore the relationship between the structure and function of the social environment upon self-reported health and (2) test the hypothesis that social relations directly affect health. In a prospective analysis over a 12-month follow-up period, we found that a lack of social support and dissatisfaction with social relations are predictive of poor health status, while weak social networks are not. Thus, functional aspects of social relations are better predictors of poor health than the structure of social interactions. Low social support was an independent risk factor for poor health in men and women: for men the effect was strongest among individuals who held a high occupational status, for women among those in high and low occupational groups. This study suggests that in France social relations exert an independent effect on health, modified by gender and socio-economic factors. Our results indicate that both clinicians in their practice and researchers may do well to focus increased attention on the health impact of social support. PMID- 12639599 TI - Context effects on texture border localization bias. AB - Observers are able to locate precisely a border defined by changes in texture orientation. The prevailing theory is that such localization takes place using a hierarchical, filter-rectify-filter mechanism. An alternative theory is that contextual modulation causes the border elements to stand out. Here we show that perceived border location is inconsistent with contextual modulation from iso oriented elements. The perceived location of a vertical border defined by vertical texture on one side, and horizontal texture on the other side, is biased towards the vertical texture. We found the same bias in a single row of texture. Therefore, the bias is not due to contextual influences from surrounding iso oriented elements. Contextual influences between cross-oriented elements can explain the data. PMID- 12639600 TI - Sustained deviation of gaze direction can affect "inverted vection" induced by the foreground motion. AB - A slowly moving foreground with an orthogonally moving background can induce self motion perception in the same direction as the foreground motion (inverted vection; [Vision Research 40 (2000) 2915]). In the present study, we investigate the effect of sustained gaze deviation on inverted vection. We hypothesized that gaze deviation affects eye-movement information registered in the perceptual system, which might be a primary factor for causing inverted vection. The experiment revealed that strength of inverted vection decreases with observer's gaze deviation in the same direction as the foreground motion, while it increases with the deviation in the opposite direction to the foreground. These results support our hypothesis and suggest that inverted vection is affected by eye movement information. PMID- 12639601 TI - Failure of signed chromatic apparent motion with luminance masking. AB - It has been suggested that there are two types of chromatic motion mechanisms: signed chromatic motion, in which correspondence across successive frames is based on chromatic content of image regions, and unsigned chromatic motion based on movement of chromatically-defined borders. We investigate whether signed and unsigned red-green chromatic motion are mediated by a genuinely chromatic mechanism. Direction discrimination of signed and unsigned red-green chromatic motion were measured in the presence of a dynamic luminance masking noise. Increasing the luminance noise contrast systematically impaired signed motion, regardless of contrast and speed. This result suggests that signed red-green chromatic motion is derived from a luminance-based signal, rather than a genuinely chromatic motion mechanism. In the case of unsigned chromatic motion, there is no effect of luminance masking noise, indicating there exists a genuine chromatic mechanism for second-order motion perception. PMID- 12639602 TI - Temporal modulation of spatial contrast vision in pigeons (Columba livia). AB - Spatiotemporal contrast sensitivity (CS) functions were obtained from four White Carneaux pigeons. The spatial frequency for each session was selected randomly from a group of five spatial frequencies ranging from 0.42 to 1.26 c/deg. Within the session, the temporal frequency varied from 1 to 32 Hz. When plotted as a function of spatial frequency, the CS functions peaked in the range 0.7-1.0 c/deg. When compared to data that had been collected at 0 Hz temporal modulation, the temporally modulated spatial CS functions showed reduced CS, especially at the higher spatial frequencies, and reduced peak spatial frequency. When plotted as a function of temporal frequency, the CS functions were flat up to 8-16 Hz. Above 16 Hz, the curves showed a sharp roll off. When plotted as a three dimensional, spatiotemporal CS surface, the data had a number of characteristics in common with the three-dimensional spatiotemporal model of CS proposed by Burbeck and Kelly (J. Opt. Soc. Am. 70 (1980) 1121). PMID- 12639603 TI - Scaling of letter size and contrast equalises perception across eccentricities and set sizes. AB - Double E(2)N(2) scaling, i.e. magnifying size and contrast, allows modelling of the deterioration of face recognition performance with increasing eccentricity (E) and the size (N) of the set from which a target face has to be identified. E(2) and N(2) values represent the eccentricities and set sizes at which stimulus size and contrast must double in order to keep performance unchanged, whilst parameter K represents the multiplicative interaction between E and N. In the current study we investigated whether double E(2)N(2) scaling can model performance deterioration with increasing eccentricity and set size in letter perception too. Contrast sensitivity for letter perception was investigated as a function of letter size at N=1-8 and E=0 degrees -10 degrees. The superimposition of contrast sensitivity functions produced two scaling surfaces, one for letter size and another for contrast, which allowed modelling of the changes in letter perception with increasing E and N. With increasing eccentricity/set size the change of scale was much faster for contrast than letter size. Thus, in letter perception, contrast scaling was more important than spatial scaling. When compared with face perception, the change of spatial scale with increasing eccentricity was slower for letters whereas the change of contrast scale was similar for both. With increasing set size the changes of both spatial and contrast scales are faster for faces. In spatial scaling the interaction between eccentricity and set size was similar for letters and faces whereas in contrast scaling letters showed no interaction. Thus, letter perception was less affected by eccentricity and set size than face perception. PMID- 12639604 TI - FliMax, a novel stimulus device for panoramic and highspeed presentation of behaviourally generated optic flow. AB - A high-speed panoramic visual stimulation device is introduced which is suitable to analyse visual interneurons during stimulation with rapid image displacements as experienced by fast moving animals. The responses of an identified motion sensitive neuron in the visual system of the blowfly to behaviourally generated image sequences are very complex and hard to predict from the established input circuitry of the neuron. This finding suggests that the computational significance of visual interneurons can only be assessed if they are characterised not only by conventional stimuli as are often used for systems analysis, but also by behaviourally relevant input. PMID- 12639605 TI - Visual field defects in albino ferrets (Mustela putorius furo). AB - The extent of the horizontal visual field was determined behaviourally in 4 pigmented and 5 albino ferrets (Mustela putorius furo, Carnivora, Mammalia) using perimetry. During binocular vision, all pigmented and three albino ferrets responded equally well to stimuli presented anywhere along the horizontal perimeter in the central 180 degrees of the visual field. The remaining two albinos had a visual field defect in the right hemifield (>30 degrees eccentricity). During monocular vision, a significant difference between the visual fields of pigmented and albino ferrets became apparent. In pigmented ferrets, the visual field of each eye included the ipsilateral (temporal) and a substantial portion of the contralateral (nasal) hemifield. In albinos, the visual field of each eye was limited to the ipsilateral hemifield and reactions to visual stimuli abruptly declined directly beyond the vertical meridian. PMID- 12639606 TI - A laminar cortical model of stereopsis and three-dimensional surface perception. AB - A laminar cortical model of stereopsis and later stages of 3D surface perception is developed and simulated. The model describes how initial stages of monocular and binocular oriented filtering interact with later stages of 3D boundary formation and surface filling-in in the lateral geniculate nucleus and cortical areas V1, V2, and V4. In particular, it details how interactions between layers 4, 3B, and 2/3A in V1 and V2 contribute to stereopsis, and clarifies how binocular and monocular information combine to form 3D boundary and surface representations. Along the way, the model modifies and significantly extends the disparity energy model. Neural explanations are given for psychophysical data concerning: contrast variations of dichoptic masking and the correspondence problem, the effect of interocular contrast differences on stereoacuity, Panum's limiting case, the Venetian blind illusion, stereopsis with polarity-reversed stereograms, da Vinci stereopsis, and various lightness illusions. By relating physiology to psychophysics, the model provides new functional insights and predictions about laminar cortical architecture. PMID- 12639607 TI - Mixture models and the probabilistic structure of depth cues. AB - Monocular cues to depth derive their informativeness from a combination of perspective projection and prior constraints on the way scenes in the world are structured. For many cues, the appropriate priors are best described as mixture models, each of which characterizes a different category of objects, surfaces, or scenes. This paper provides a Bayesian analysis of the resulting model selection problem, showing how the mixed structure of priors creates the potential for non linear, cooperative interactions between cues and how the information provided by a single cue can effectively determine the appropriate constraint to apply to a given image. The analysis also leads to a number of psychophysically testable predictions. We test these predictions by applying the framework to the problem of perceiving planar surface orientation from texture. A number of psychophysical experiments are described that show that the visual system is biased to interpret textures as isotropic, but that when sufficient image data is available, the system effectively turns off the isotropy constraint and interprets texture information using only a homogeneity assumption. Human performance is qualitatively similar to an optimal estimator that assumes a mixed prior on surface textures--some proportion being isotropic and homogeneous and some proportion being anisotropic and homogeneous. PMID- 12639609 TI - Can DHEA be used to prevent bone loss and osteoporosis? PMID- 12639610 TI - Is degenerative disk disease genetically determined? PMID- 12639611 TI - Parvovirus B19 and autoimmune diseases. AB - Parvovirus B19 (B19) causes many clinical disorders, of which the most common are erythema infectiosum, aplastic crisis complicating chronic hemolytic anemia, and hydrops fetalis. In young adults, the skin eruption caused by B19 is accompanied with polyarthritis and polyarthralgia in 60% of the cases. The joint abnormalities predominate in the hands and feet and usually resolve within a week (range 2-21 d). Serological tests show IgM antibodies against B19, confirming the diagnosis of recent infection. Protracted polyarthritis occurs in some patients and seems associated with the DR4 histocompatibility alleles. Rheumatoid factors can be produced transiently in these patients. Other autoantibodies produced in the wake of B19 infection include anti-nuclear antibodies, anti-DNA, anti SSA/SSB, and anti-phospholipids. Acute B19 infection can simulate early rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE) (lupus-like eruption over the cheeks, cytopenia, etc.). In addition, there have been a few reports of erosive RA or SLE developing shortly after a B19 infection, with positive PCR tests for B19 DNA in synovial tissue or blood cells. Studies in large series indicate that B19 is probably an extremely rare cause of RA or SLE. Vasculitides affecting the small vessels (Henoch-Schonlein purpura, Wegener's granulomatosis), medium-sized vessels (periarteritis nodosa), and large vessels (giant cell arteritis) can occur after B19 infection. Here again, the number of clinical cases is small. PMID- 12639612 TI - Reflex sympathetic dystrophy syndrome and neuromediators. AB - Concepts related to the pathophysiology of reflex sympathetic dystrophy syndrome (RSDS) are changing. Although sympathetic influences are still viewed as the most likely mechanism underlying the development and/or perpetuation of RSDS, these influences are no longer ascribed to an increase in sympathetic tone. Rather, the most likely mechanism may be increased sensitivity to catecholamines due to sympathetic denervation with an increase in the number and/or sensitivity of peripheral axonal adrenoceptors. Several other pathophysiological mechanisms have been suggested, including neurogenic inflammation with the release of neuropeptides by primary nociceptive afferents and sympathetic efferents. These neuromediators, particularly substance P, calcitonin gene-related peptide, and neuropeptide Y (NPY), may play a pivotal role in the genesis of pain in RSDS. They induce an inflammatory response (cutaneous erythema and edema) and lower the pain threshold. Neurogenic inflammation at the site of the lesion with neuromediator accumulation or depletion probably contributes to the pathophysiology of RSDS. However, no single neuromediator has been proved responsible, and other hypotheses continue to arouse interest. PMID- 12639613 TI - Leptin: a potential mediator for protective effects of fat mass on bone tissue. AB - Body weight is among the most powerful predictors of bone status, and adipose tissue plays a substantial role in weight-related protective effects on bone. An understanding of the mechanisms underlying the relation between adipose tissue and bone may open up new perspectives for treatment. Leptin, which is known to regulate appetite and energy expenditures, may also contribute to mediate the effects of fat mass on bone. Although reported data are somewhat conflicting, there is some evidence that leptin may decrease bone formation via a central nervous effect and may stimulate both bone formation and bone resorption via direct peripheral effects on stromal precursor cells. The net result of these central and peripheral effects may depend on serum leptin levels and blood-brain barrier permeability, of which the first increase and the second decrease as obesity develops. Further work is needed to improve our understanding of these effects. PMID- 12639614 TI - Total knee arthroplasty in hemophilic arthropathy. AB - Chronic arthropathy causes major functional disability in patients with severe hemophilia. OBJECTIVE: To evaluate the results of total knee arthroplasty (TKA) and its impact on both quality of life and clotting factor use in patients with severe hemophilia. PATIENTS AND METHODS: We evaluated 17 TKAs in 12 patients. The TKAs were performed between 1986 and 1996, and follow-up was 8-132 months (mean, 54 months). Mean age at arthroplasty was 39 years (22-51 years). Quality of life was evaluated using the Short Form 36 (SF-36). RESULTS: Results were good or excellent in 94% of patients. The improvement was greatest for pain. Recurrent hemarthrosis in six patients and development of an anticoagulant in two patients were the only postoperative complications. Clotting factor use did not decrease significantly after surgery. SF-36 scores showed an increase in physical activity responsible for an improvement in quality-of-life indicators. However, this improvement in functional capabilities seemed to wane over time as a result of arthropathy in other joints and of intercurrent diseases. CONCLUSION: TKA for hemophilic arthropathy provides good results that translate into quality-of-life gains. PMID- 12639615 TI - Temporal arteritis and polymyalgia rheumatica in north-eastern Spain: clinical spectrum and relationship over a 15 year period. AB - OBJECTIVE: To examine the clinical spectrum of polymyalgia rheumatica (PMR) and temporal arteritis (TA) and their relationship over a period of 15 years in an area of north-eastern Spain. METHODS: We undertook a descriptive study of an unselected population of 163 patients with PMR and/or TA diagnosed from 1985 to 1999. RESULTS: Of the 163 patients included, 90 had isolated PMR, 41 had PMR associated with TA, and 32 had isolated TA. The clinical spectrum of both conditions in our area was similar to that reported in other populations, including a marked female predominance. However, in our series, no patient developed permanent blindness or other major ischemic complications. PMR was observed in 56% of patients with TA. Conversely, 7% of patients originally suffering from PMR without clinical evidence of arteritis at presentation developed later symptoms of TA, and there were no predictive features for this. Interestingly, none of these patients suffered visual loss or other ischemic complications. The low risk of major complications in these cases does not support the need for systematic arterial biopsy in all patients with symptoms of PMR alone. On comparing patients with isolated TA with patients with PMR associated with TA, no differences were observed, thus discarding the possibility that the second constitutes a distinct and independent subgroup of TA. In contrast, when comparing patients with isolated PMR with patients with PMR associated with TA, we found significant differences between both the groups, with greater abnormality of clinical and laboratory markers of inflammation in patients with PMR associated with TA. These differences seem to reflect a greater degree of systemic inflammation linked to the presence of TA. CONCLUSION: In our area, TA appears nowadays as a benign disease which infrequently presents blindness or other major complications. Our experience confirms that even after a good clinical response with normalization of a high ESR in PMR, the patient is at risk for clinical development of TA. Finally, our study also shows that isolated TA and PMR associated with TA seem to be the same condition, different from isolated PMR. PMID- 12639616 TI - Bone mineral density in systemic lupus erythematosus and its relation to age at disease onset, plasmatic estradiol and immunosuppressive therapy. AB - OBJECTIVE: The aim of this paper was to evaluate bone mineral density (BMD) in patients with systemic lupus erythematosus (SLE), to determine the role of corticosteroids and cytotoxic drugs and to assess estrogen effect on BMD in SLE. PATIENTS AND METHODS: BMD (DEXA) at lumbar vertebrae (L2-L4) and at femoral neck was performed in 60 pre-menopausal SLE patients and in 64 controls. Estradiol level was measured in all the individuals. Age, age at disease onset, body mass index (BMI), time of disease, disease activity (SLEDAI), prednisone dose at the evaluation, total cumulative and cumulative prednisone dose in the last year and cytotoxic drugs were assessed. RESULTS: The mean plasmatic estradiol was 175.9 pg/ml in patients and 149.9 in controls. BMD was inferior in patients than that in controls (P < 0.0001). The mean current, cumulative and previous year prednisone doses were, respectively, 19.17 mg/d, 28.78 g and 5.33 g. There was no association between corticosteroids or cytotoxic drug used and low bone mass. The serum concentration of estradiol did not influence the bone mass. The BMI and age at disease onset exhibited an influence on BMD at L2. CONCLUSIONS: BMD was significantly lower in SLE patients but not related to CS (Corticosteroids)or other drugs; the estradiol in these patients had no effect on BMD. Low BMI interacting with early onset of disease might influence the probability of loss of bone mass. PMID- 12639617 TI - Is ultrasound of bone relevant for corticosteroid-treated patients? A comparative study with bone densitometry measured by DEXA. AB - Corticosteroid treatment diminishes bone mass and alters bone quality. The objective was to evaluate bone in corticosteroid-treated patients and controls and in fractured and non-fractured patients treated with corticosteroids using both X-ray densitometry (DEXA) and ultrasound. We evaluated 34 women aged 58 +/- 14 years (X +/- SD), who had been on long-term low dose prednisone therapy for at least 6 months, and who had never received specific treatment for osteoporosis. Bone mineral density of total skeleton (TS), lumbar spine (LS), femoral neck (FN), and vertebral morphometry (MXA) were measured by DEXA. Speed of sound (SOS), broadband ultrasound attenuation (BUA) and stiffness were measured using an Achilles Plus system. Forty-two healthy women served as controls. Both densitometric and ultrasound parameters in the patients were significantly diminished compared with controls: TS: P < 0.002, LS: P < 0.025, FS: P < 0.005, Stiffness: P < 0.001, BUA: P < 0.002 and SOS: P < 0.002. The percentage of patients with a Z score below -2 was higher in Stiffness and BUA: 38% and 47%, respectively, compared with a range of 16-24% in the other parameters (P < 0.05 BUA vs. DEXA measurements). Eleven patients with previous bone fracture had values lower than the non-fractured patients, both according to DEXA and ultrasound measurements, but the difference was only significant for BUA (P < 0.02). BUA of the calcaneus was more effective in detecting the specific skeletal alterations and fracture risk of the group of patients receiving chronic corticosteroid treatment. PMID- 12639618 TI - Informing patients about serious side effects of drugs. A 2001 survey of 341 French rheumatologists. AB - OBJECTIVES: In May 2001, we surveyed French rheumatologists to determine how they inform their patients about serious side effects of drugs. RESULTS: Of the 600 rheumatologists invited to participate in the study, 341 completed and returned the questionnaire. Most respondents believed that full information on patients would become the rule (mean, 62.1 +/- 28.3, the items being scored on a 100-point scale where 0 was "not at all" and 100 "absolutely"). The answer to this item varied little across age groups. Respondents felt that patient information fell primarily on the physician, (78 +/- 23), followed by pharmaceutical companies (64 +/- 32) and pharmacists (46 +/- 33). The respondents believed that full information of patients about side effects would primarily benefit physicians (51 +/- 32), followed by pharmaceutical companies (43 +/- 30) and patients (38 +/- 28). The reaction of patients to information on drug-related risks was described as "always favorable" by 2% of the respondents and as "fairly often favorable" by 55%. Among respondents, 9% always gave information on serious side effects, 66% fairly often, and 4% never; 2% always communicated information on the approximate frequency of serious adverse events, 21% fairly often, and 33% never. Similarly, 3% of the respondents always warned their patients of lethal risks, 11% fairly often, and 44% never. When evaluated on a 100-point scale (0, not at all knowledgeable; 100, very knowledgeable), knowledge of serious side effects of drugs for musculoskeletal conditions was 73 +/- 23, whereas knowledge of the frequency of these effects was only 52 +/- 22. Eight percent of the respondents always recommended that patients read the package insert, 27% fairly often, and 32% never. Conversely, the possible adverse effects of not taking the treatment were always specified by 21% of the respondents and fairly often by 88%. PMID- 12639619 TI - Recurrent hemarthrosis following total knee arthroplasty. Report of a case treated with arterial embolization. AB - Recurrent spontaneous hemarthrosis of the knee is common and often related to a detectable cause. In contrast, hemarthrosis after knee arthroplasty is rare and frequently unexplained. We report a case of recurrent hemarthrosis 6 years after total knee arthroplasty. The cause was hypervascularization about the joint, and embolization therapy was effective. PMID- 12639620 TI - Osteitis pubis after Marshall-Marchetti-Krantz urethropexy. AB - We report a case of osteitis pubis after Marshall-Marchetti-Krantz urethropexy. There are several theories on the pathogenesis of the postoperative osteitis pubis and inconsistent results with a wide variety of treatments. Also, osteomyelitis and osteitis pubis can have identical symptoms. In our case, the use of a Mitek anchor system and its migration within the joint cavity might have induced a foreign-body reaction as the responsible mechanism of the joint destruction. PMID- 12639622 TI - Giant intraosseous cyst-like lesions in rheumatoid arthritis report of a case. AB - The term "intraosseous synovial cyst" is used to designate both the epiphyseal cyst-like lesions seen in patients with rheumatoid arthritis (RA) and mucoid cysts, which occur in a different setting. We report the case of a patient in whom a 4-cm cyst-like lesion developed in the left tibia 18 years after onset of RA and 6 years after osmic acid synovectomy of the left knee. Positive contrast arthrography and magnetic resonance imaging visualized a communication between the lesion and the joint space. Preexisting bone and joint lesions and increased intraarticular pressure play a major role in the genesis of cyst-like lesions in RA. In our patient, the osmic acid synovectomy may have contributed to the development of the lesion. "Synovial cyst" is a misnomer for these giant lesions, which are geodes rather than cysts. Despite their low incidence, these lesions deserve attention because they raise diagnostic and therapeutic problems. PMID- 12639621 TI - Aggressive systemic mastocytosis. AB - Systemic mastocytosis is a rare and occasionally aggressive condition that raises major diagnostic challenges. We report a case in a 72-year-old patient in whom the diagnosis of malignant mastocytosis required two bone marrow smears and three bone marrow biopsies examined using specific staining techniques. Despite interferon therapy, a mast-cell sarcoma of the sternum developed 1 year after symptom onset, followed 1 year later by acute myeloblastic leukemia, which was rapidly fatal. PMID- 12639623 TI - Fasciola hepatica infestation with joint symptoms. PMID- 12639624 TI - A case of cutaneous sensory neuropathy associated with Churg-Strauss syndrome. AB - Cutaneous sensory neuropathy manifests as multiple, sharply demarcated areas of hypoesthesia with a variable degree of pain. This rare neuropathy is caused by a multifocal infection or inflammation of the small sensory nerves of the skin. We report a case in a patient with febrile arthritis and eosinophilia. Her chronic cough and the presence of extravascular infiltrates of eosinophils in a neuromuscular biopsy specimen suggested Churg-Strauss syndrome. The course was favorable under corticosteroid therapy. PMID- 12639626 TI - Andrew Taylor Still and the birth of osteopathy (Baldwin, Kansas, USA, 1855). AB - Osteopathy has gained ground in recent years and has been seeking recognition in France. Physicians often lack the information needed to answer patients who have derived from the media, advertisements, and other patients what they believe is a clear idea of osteopathy and its twin sister chiropractic. My academic activities led me to the heart of the United States, to Kansas, where settlers and Indians once stood face to face and where wagon trains left daily for the Western territories. There, in Baldwin, Andrew Taylor Still "discovered" osteopathy. I conducted an in-depth study of the birth of osteopathy and of the ideological and cultural influences that shaped this doctrine. The circumstances that surrounded the development of osteopathy deserve to be widely known because they explain how contemporary osteopaths work. Indeed, although the terms are different, the ideology that underlies osteopathy seems unchanged. The history of osteopathy emphasizes the importance of logical thinking in medicine, of the principle of pathophysiological foundation, of diagnostic hypotheses, and of careful treatment selection complying with the rules of deontology and ethics. Osteopathy is without doubt a product of society and perhaps also of vogue. It cannot leave physicians indifferent. PMID- 12639625 TI - Anterior tibial compartment syndrome due to the pyomyositis in a patient with rheumatoid arthritis. A case report. AB - Anterior tibial compartment syndrome was developed due to pyomyositis in a 33 year-old male patient with rheumatoid arthritis while receiving steroid therapy during the follow-up period. The preoperative physical examination, laboratory findings, MRI images, intraoperative observation and postoperative histopathological examinations confirmed the association with pyomyositis. The surgical drainage and antibiotic treatment were effective, and in the follow-up period, neuromuscular dysfunctions disappeared completely within 6 months. The patient has been asymptomatic for 4 years of follow-up. To date, anterior tibial compartment syndrome due to pyomyositis in a case with rheumatoid arthritis has not been reported. PMID- 12639627 TI - Sciatic nerve palsy as presenting sign of a perianal abscess. PMID- 12639628 TI - Upper extremity contractures heralding Parkinson's disease. PMID- 12639629 TI - Insufficiency fracture of the sternum report of a case. PMID- 12639630 TI - Rationale for cavernous nerve restorative therapy to preserve erectile function after radical prostatectomy. PMID- 12639631 TI - Initial evaluation of Cyberknife technology for extracorporeal renal tissue ablation. AB - OBJECTIVES: To determine whether Cyberknife technology can be applied to renal tissue safely and effectively. The goal was to achieve the high efficacy of a surgical treatment, with the low morbidity of a noninvasive intervention. METHODS: The Cyberknife is a frameless, image-guided radiosurgical device. This innovative extracorporeal treatment combines a linear accelerator mounted on a highly maneuverable robotic arm. The Cyberknife is unique in that it divides the high-dose radiation necessary to ablate the lesion completely into up to 1200 beams. Each one of these beams of radiation has a significantly reduced dose. Therefore, the individual dose of each beam is essentially benign to the pathway and surrounding tissue. However, at the focal point of these beams, the dose is additive, and the desired ablative dose is attained. Predetermined "lesions" in 16 kidneys were treated in vivo in the porcine model. Complete treatment was accomplished in one session per animal, with no complications. Gross and histologic evaluations were completed at 4, 6, or 8 weeks. RESULTS: The degree of radiation changes correlated with longer treatment intervals. After 8 weeks, the lesions showed complete fibrosis. The zones of complete fibrosis were characterized by dense, paucicellular connective tissue completely devoid of all normal kidney elements, including tubules and glomeruli. CONCLUSIONS: This initial preclinical evaluation of the Cyberknife for extracorporeal renal tissue ablation appears to be very promising and demonstrated its ability to ablate a targeted area precisely and completely with relative sparing of the surrounding tissue. This innovative technology introduces an exciting approach as a potential treatment option of renal masses in the future. PMID- 12639632 TI - Long-term results for Malone antegrade continence enema for adults with neurogenic bowel disease. AB - OBJECTIVES: To determine the long-term outcomes from the Malone antegrade continence enema (ACE) procedure in adult neurogenic patients. METHODS: A retrospective review of adult patients who underwent the ACE procedure for neurogenic bowel was done. Patients were studied if they had follow-up of greater than 4 years. We compared pre-ACE and post-ACE toileting times, bowel continence status, and complications, and elicited patient subjective satisfaction with their quality of life. RESULTS: Six patients were available for study. Mean age was 35 years with a mean follow-up of 4.5 years. Urinary diversion was done in 5 patients. Pre-ACE toileting time was 190 +/- 45 minutes versus post-ACE toileting time of 28 +/- 20 minutes (P <0.001). Four of six patients pre-ACE were incontinent of stool per rectum compared with 1 of these 4 patients post-ACE (P = 0.03). Five patients were continent of stool per ACE stoma. Four patients (67%) had complications. Three of five patients (60%) who underwent synchronous urinary diversion required postoperative re-exploration. Five patients (83%) were satisfied with their outcome and rated their quality of life higher after the ACE procedure compared with pre-ACE. CONCLUSIONS: The ACE procedure is effective in the long-term management of adult neurogenic bowel. The complication and re exploration rates are high. Patients must be properly selected to determine appropriate motivation. PMID- 12639633 TI - Comparison of red blood cell volume distribution curves and phase-contrast microscopy in localization of the origin of hematuria. AB - OBJECTIVES: To determine the ability, reliability, and accuracy of urinary red blood cell volume distribution curves obtained with the use of an autoanalyzer to identify the origin of isolated microscopic hematuria and compare the results with those obtained with phase-contrast microscopy (PCM). METHODS: A prospective evaluation was performed in 45 patients with glomerular or nonglomerular microhematuria detected by urinalysis, PCM, radiologic evaluation, endoscopy, and, sometimes, renal biopsy. Urine samples were analyzed in an electronic particle-size analyzer, and the tests were repeated to assess reliability. The kappa correlation coefficient was used to assess reliability and to compare the results with the final diagnosis and with those obtained with PCM. RESULTS: Of the 28 patients who had a single definite cause of hematuria, 16 had glomerular bleeding and 12 nonglomerular bleeding. The origin of hematuria was correctly identified by the autoanalyzer in 60.7% of cases. A statistically significant correlation was found with the final diagnosis (kappa = 0.433, P = 0.048). The reliability was excellent (kappa = 0.917, P <0.0001). Of 16 patients with glomerulonephritis, 10 (62.5%) were correctly identified by PCM and 14 (87.5%) by the autoanalyzer. In 12 patients with nonglomerular bleeding, PCM was accurate in 7 (58%) and the autoanalyzer in 3 (25%). The results were statistically correlated with the findings of PCM (kappa = 0.327, P <0.00001). CONCLUSIONS: The use of an autoanalyzer is easy, reproducible, and noninvasive. It provides reliable information to orient the diagnosis toward glomerular or nonglomerular bleeding. It is as accurate as PCM for screening for the source of hematuria. PMID- 12639634 TI - Cold-knife endoureterotomy for nonmalignant ureterointestinal anastomotic strictures. AB - OBJECTIVES: To evaluate the long-term results of cold-knife incision (CNI) of nonmalignant ureterointestinal anastomosis strictures (UASs) after urinary diversion in a consecutive series of patients. METHODS: Since 1994, we have evaluated retrospectively 40 patients with 43 UASs, who were primarily treated with CNI (group 1). Six patients from group 1 with 7 UASs who failed primary CNI comprised group 2. After placement of an 8F nephrostomy tube, a 0.035-inch guidewire bypassed the stricture in an antegrade fashion under guidance of a centrally opened ureteral catheter (5F). A wire-mounted cold-knife was pulled through the strictured area in retrograde fashion under fluoroscopic control. Postoperatively, an 8 to 12F stent was left indwelling for 6 to 12 weeks. Successful treatment was defined as radiographic and scintigraphic resolution of obstruction and symptomatic relief. RESULTS: In group 1, after removal of the stent, the ureteroenteric area remained patent in 26 (60.5%) of 43 UASs during a follow-up period of 38.8 months (range 12 to 85). The success rate at 1, 2, and 3 years was 86%, 67.8%, and 60.5%, respectively. In group 2, no success occurred. The diameter and length of the stricture, kidney function, hydronephrosis grade, presence of urinary infection at presentation, past CNI or radiotherapy, number of incisions with the cold-knife, and premature appearance of the anastomosis stricture were statistically significant influences on the outcome (P <0.05). Considering only the patients (n = 8) with the most favorable predictive factors (interval to stricture formation 12 months or longer, stricture length 1.5 cm or less, and hydronephrosis grade I-II), the success rate was 100%. No complications were observed. CONCLUSIONS: CNI is an effective and minimally invasive treatment for primary UASs, providing durable results compared with other modalities used for endoureterotomy, and should be considered as an initial approach. The selection of patients with the most favorable prognostic factors leads to excellent results. As a secondary procedure, CNI was not successful. PMID- 12639636 TI - Assessment of stricture formation with the ureteral access sheath. AB - OBJECTIVES: To analyze the long-term incidence of ureteral stricture formation in a series of patients in whom a new-generation ureteral access sheath was used. A new generation of ureteral access sheaths has been developed to facilitate ureteroscopic procedures. However, some have questioned their safety and whether the device might cause significant ureteral trauma. METHODS: Between September 1999 and July 2001, 150 consecutive ureteroscopic procedures with adjunctive use of an access sheath were performed. A retrospective chart review to April 2002 was done. Of the 150 patients, 130 underwent ureteroscopy for ureteral stones. Patients who underwent endoureterotomy or treatment of transitional cell carcinoma were excluded from this analysis. Sixty-two patients had follow-up greater than 3 months and were included in the analysis. Overall, 71 ureteroscopic procedures were performed, with 9 patients undergoing multiple procedures. Ninety-two percent of the patients had pathologic findings above the iliac vessels. The average patient age was 45.3 years (range 17 to 76), and 70% and 30% of the patients were male and female, respectively. The mean clinical follow-up was 332 days (range 95 to 821), and follow-up imaging was performed within 3 months after ureteroscopy in all patients. RESULTS: The 10/12F access sheath was used in 8 ureteroscopic procedures (11.2%), the 12/14F access sheath in 56 (78.9%), and the 14/16F access sheath in 7 (9.8%). One stricture was identified on follow-up imaging of 71 procedures performed, for an incidence of 1.4%. The patient developed the stricture at the ureteropelvic junction after multiple ureteroscopic procedures to manage recurrent struvite calculi. The access sheath did not appear to be a contributing factor. CONCLUSIONS: The results of our series indicate that the ureteral access sheath is safe and beneficial for routine use to facilitate flexible ureteroscopy. However, awareness of the potential ischemic effects with the use of unnecessarily large sheaths for long periods in patients at risk of ischemic injury should be considered. We advocate the routine use of the device for most flexible ureteroscopic procedures proximal to the iliac vessels. PMID- 12639640 TI - Improved detection of renal pathologic features on multiphasic helical CT compared with IVU in patients presenting with microscopic hematuria. AB - OBJECTIVES: To examine the virtues of multiphasic helical computed tomography (CT) in the diagnosis of upper urinary tract lesions refractory to identification by intravenous urography (IVU). METHODS: A total of 86 patients (59 men and 27 women), 27 to 88 years old, with microscopic hematuria and negative IVU findings were examined with multiphasic helical CT consisting of a pre-enhancement, late arterial-early cortical-medullary, nephrographic, and excretory phase helical CT of the kidneys, using 3 to 5-mm collimation and 7.5-mm/s table feed. RESULTS: The multiphasic helical CT was conclusive in 84 lesions. Twenty-five cases of early papillary and medullary necrosis, 7 of 8 inflammatory lesions, 3 caliceal diverticula, 1 lupus nephritis, 26 small calculi, 2 medullary sponge kidney, 5 vascular anomalies, and 3 infarcts presented with characteristic manifestations on CT but lacked findings on IVU. Similarly 8 of 9 small malignant neoplasms, 2 small benign neoplasms, and 2 small cysts produced no detectable findings on IVU but were readily diagnosed on helical CT. CONCLUSIONS: Characteristic findings, particularly on late arterial, early corticomedullary, parenchymal, and excretory phase helical CT make possible the diagnosis of early inflammatory disease, small masses and neoplastic lesions, and vascular abnormalities. Limited resolution (needed to identify small calculi) and the lack of ability to capture phases significantly reduce the diagnostic ability of IVU. PMID- 12639639 TI - Biochemical profile of stone-forming patients with diabetes mellitus. AB - OBJECTIVES: To test the hypothesis that stone-forming patients with type II diabetes (DM-II) have a high prevalence of uric acid (UA) stones and present with some of the biochemical features of gouty diathesis (GD). METHODS: The demographic and initial biochemical data from 59 stone-forming patients with DM II (serum glucose greater than 126 mg/dL, no insulin therapy, older than 35 years of age) from Dallas, Texas and Durham, North Carolina were retrieved and compared with data from 58 patients with GD and 116 with hyperuricosuric calcium oxalate urolithiasis (HUCU) without DM. RESULTS: UA stones were detected in 33.9% of patients with DM-II compared with 6.2% of stone-forming patients without DM (P <0.001). Despite similar ingestion of alkali, the urinary pH in patients with DM II and UA stones (n = 20) was low (pH = 5.5), as it is in patients with GD, and was significantly lower than in patients with HUCU. The urinary pH in patients with DM-II and calcium stones (n = 39) was intermediate between that in those with DM-II and UA stones and those with HUCU. However, both DM groups had fractional excretion of urate that was not depressed, as it is in those with GD, and was comparable to the value obtained in those with HUCU. The urinary content of undissociated UA was significantly higher, and the saturation of calcium phosphate (brushite) and sodium urate was significantly lower in those with DM-II and UA stones than in those with HUCU. CONCLUSIONS: Stone-forming patients with DM-II have a high prevalence of UA stones. Diabetic patients with UA stones share a key feature of those with GD, namely the passage of unusually acid urine, but not the low fractional excretion of urate. PMID- 12639641 TI - Retroperitoneoscopic nephroureterectomy for transitional cell carcinoma of the renal pelvis and ureter: Nagoya experience. AB - OBJECTIVES: To evaluate the efficacy of our new retroperitoneoscopic nephroureterectomy for patients with transitional cell carcinoma of the renal pelvis and ureter, we present the operative procedure and analysis of the clinical outcome of retroperitoneoscopy in 23 patients. METHODS: Twenty-three patients with transitional cell carcinoma of the upper urinary tract underwent retroperitoneal laparoscopic nephroureterectomy between February 2000 and February 2002. Patient age ranged from 44 to 83 years (mean 66.7). Each kidney was retroperitoneoscopically dissected en bloc, together with the perirenal fatty tissue, lymph nodes, and/or adrenal gland, without transecting the ureter. The lower ureter was resected with the bladder cuff transected using an ultrasonic scalpel and an endoscopic gastrointestinal automatic stapler. The dissected specimen was removed intact through a 6-cm-long original incision. RESULTS: The mean operating time was 4.8 hours, including 0.7 hours for complete removal of the ureteral end with the bladder cuff. The mean estimated blood loss was 304 mL. The mean time to recovery to normal activity was 18 days. In the mean follow-up period of 15 months, 2 patients died of cancer progression in the sixth postoperative month, 2 died of other causes, and 4 had recurrent transitional cell carcinoma of the bladder after surgery. CONCLUSIONS: Our retroperitoneal laparoscopic nephroureterectomy using an endoscopic gastrointestinal automatic stapler is a fast, low-risk, and minimally invasive procedure and might be an alternative to other laparoscopic techniques and open nephroureterectomy. However, long-term follow-up is necessary to confirm the efficacy for patients with transitional cell carcinoma of the renal pelvis and ureter. PMID- 12639642 TI - Evaluation of DD23 as a marker for detection of recurrent transitional cell carcinoma of the bladder in patients with a history of bladder cancer. AB - OBJECTIVES: To determine whether DD23 increases the sensitivity of urinary-based detection of transitional cell carcinoma (TCC) recurrence. The murine monoclonal antibody DD23 recognizes a 185-kDa tumor-associated antigen that is expressed in human bladder cancer cells in vitro and in vivo but is not detected in normal urothelium. METHODS: Using alcohol-fixed urinary cytology, matched voided urine and bladder wash specimens were evaluated for the contribution of DD23 antigen expression in the detection of recurrent TCC. The selected patient population had a history of bladder cancer, and urine cytology analysis was performed in a single commercial reference laboratory. DD23 antigen expression in a cohort of 81 patients was compared with urine cytology findings, and the sensitivity and specificity for each urine-based test was determined. The presence of recurrent disease was determined by positive pathologic biopsy. RESULTS: The 81-patient cohort produced 151 urine specimens for which both biopsy and cytology information were obtained. Of these specimens, 64 were confirmed by a tissue diagnosis for TCC recurrence. These biopsy-proven recurrences were used as the dependent variable to assess the accuracy of cytology testing. For the detection of TCC, the DD23 antigen had a sensitivity of 70.3% and a specificity of 59.8%. Combined with cytopathologic findings, DD23 enhanced the sensitivity for the detection of TCC from 43.8% (cytology alone) to 78.1%. For low-grade TCC (n = 20) DD23 enhanced the sensitivity from 20.0% (cytology alone) to 55.0%. For high grade TCC (n = 25), DD23 enhanced the sensitivity from 64.0% (cytology alone) to 76.0%. In patients with a prior history of intravesical treatment, DD23 had a sensitivity of 94.7% and a specificity of 33.3% compared with a sensitivity of 52.6% and a specificity of 83.3% for cytology. CONCLUSIONS: DD23 antigen expression can be used as an adjunct to cytopathologic evaluation to enhance the sensitivity of urinary cytology detection of TCC. In addition, DD23 does not appear to lose sensitivity in patients with a prior history of bladder cancer treated with intravesical agents. PMID- 12639643 TI - Videourodynamic analysis of the relationship of Valsalva and cough leak point pressures in women with stress urinary incontinence. AB - OBJECTIVES: To use videourodynamic assessment to investigate the relationship between Valsalva leak point pressure (VLPP) and cough leak point pressure (CLPP) in women with stress urinary incontinence (SUI). METHODS: A total of 116 women with SUI were retrospectively analyzed by videourodynamic results, urethral pressure profilometry, and leak point pressure findings. All women had urine leakage during both coughing and performance of the Valsalva maneuver. The grade of SUI, VLPP, CLPP, maximal urethral closure pressure, and bladder neck descent during the stress test were compared among patients categorized according to three VLPP groups of less than 60 cm H(2)O, 60 or greater but less than 90 cm H(2)O, and 90 cm H(2)O or greater. RESULTS: A significant inverse correlation was found between the grade of SUI and VLPP (r = -0.620, P = 0.000) and CLPP (r = 0.506, P = 0.000). The VLPP was less than 60 cm H(2)O in 41 women, 60 or greater but less than 90 cm H(2)O in 36, and 90 cm H(2)O or greater in 39. The CLPP was greater than the VLPP in 101 women (87.1%) and equaled the VLPP in 15 women (12.9%). Of the 41 women with a VLPP less than 60 cm H(2)O, 38 (92.6%) had a CLPP greater than the VLPP and 16 (39%) had greater bladder neck descent. The maximal urethral closure pressure had a weak but significant correlation with the VLPP (r = 0.299, P = 0.011) and CLPP (r = 0.325, P = 0.005). CONCLUSIONS: The results of this study have demonstrated that the Valsalva maneuver and cough caused urinary leakage by different activities. A lower leak point pressure was found with VLPP than with CLPP to diagnose intrinsic sphincter deficiency. Complete assessment of urethral function, VLPP, CLPP, and bladder neck descent is essential for diagnosis and treatment of SUI. PMID- 12639645 TI - Effect of botulinum a toxin in the treatment of voiding dysfunction due to detrusor underactivity. AB - OBJECTIVES: To investigate the effects of botulinum A toxin in treating patients with voiding dysfunction due to detrusor underactivity. METHODS: Twenty patients with chronic urinary retention (n = 13) or severe dysuria (n = 7) received 50 U of botulinum A toxin by urethral injection. The clinical effects, obstructive symptom score, quality-of-life index, and urodynamics were compared at baseline and after treatment. RESULTS: Of the 4 males and 16 females (age range 14 to 86 years) with voiding dysfunction (cauda equina lesion in 5, dysfunctional voiding in 5, peripheral neuropathy in 6, and detrusor failure of unknown origin in 4), 18 (90%) were treated satisfactorily. Among these patients, the mean quality-of life score decreased significantly from 5.68 +/- 0.67 to 1.16 +/- 1.61. The median voiding pressure (56.5 +/- 41.2 versus 39.0 +/- 38.4 cm H(2)O) decreased significantly, as did the maximal urethral closure pressure (65.5 +/- 38.1 versus 50 +/- 32.1 cm H(2)O) and residual urine volume (300 +/- 189.1 versus 50 +/- 153.6 mL) at 2 weeks after treatment and remained stationary for 3 months. The subjective maximal effect was achieved within 1 to 2 weeks. In 7 patients, the indwelling catheters were removed, and in 4 patients who performed clean intermittent self-catheterization, the frequency decreased or it was discontinued. The other 7 patients with difficult urination had significant improvement in the obstructive symptom score (18 +/- 3.3 versus 7 +/- 4.5, P = 0.000). CONCLUSIONS: Botulinum A toxin at a dose of 50 U was effective in reducing urethral sphincter resistance among our patients with detrusor underactivity and difficult urination. PMID- 12639646 TI - Intravesical ethanol test: an ineffective measure of bladder hyperpermeability. AB - OBJECTIVES: To evaluate the intravesical ethanol test as a quantitative measure of bladder hyperpermeability in patients with interstitial cystitis. METHODS: Patients with a clinical diagnosis of interstitial cystitis and a positive result on the Parsons potassium test were enrolled in the study. The intravesical ethanol test was conducted after completion of the Parsons potassium test. Dose escalation (10% to 20% ethanol) was performed in an attempt to achieve detectable levels of ethanol. Breathalyzer data were recorded in 5-minute intervals for 30 minutes using a calibrated Intox Breathalyzer Model IV Alcosensor. The bladder was then drained, and volume recovered was recorded. The breathalyzer data were recorded for an additional 30 minutes after removal of the catheter at 5-minute intervals. RESULTS: The 3 patients studied demonstrated significant qualitative evidence of bladder hyperpermeability, with pain and urgency scores ranging from 4 to 5 on the Parsons potassium test. No detectable levels of ethanol were identified in any of the 3 patients after the intravesical ethanol test, with up to 50 mL of 20% ethanol instilled for 30 minutes. Intravesical ethanol instillation was not associated with significant pain or irritative voiding symptoms. CONCLUSIONS: The intravesical ethanol test did not prove to be a useful clinical adjunct in the diagnosis and management of interstitial cystitis. The evaluation of higher concentrations of ethanol may be warranted. PMID- 12639647 TI - Conservative treatment of female urinary incontinence with functional magnetic stimulation. AB - OBJECTIVES: To determine the efficacy and safety of functional magnetic stimulation (FMS) produced by the Pulsegen device compared with placebo in the treatment of women with urinary incontinence. METHODS: Fifty-five women with urinary incontinence were randomly assigned to the active FMS group (30 patients) or the placebo group (22 patients). Each patient in the active group received a Pulsegen device, which produced a pulsating magnetic field of B = 10 microT intensity and a frequency of 10 Hz. Patients were asked to wear the Pulsegen device day and night for 2 months. Clinical and urodynamic data were collected before and after FMS and analyzed using nonparametric statistics. RESULTS: Compared with the placebo, the number of pads used was significantly lower (P = 0.0031) after FMS, as was the pad weight (P = 0.014). In patients from the active group, a significant improvement in the power of the pelvic floor muscle contractions (P = 0.0071), as well as in the duration of the pelvic floor muscle contractions (P = 0.038), was observed. After FMS, a 56.3% improvement in urinary incontinence symptoms was reported by patients in the active group, a significantly greater difference (P = 0.00012) compared with the reported 26.3% improvement in symptoms in the placebo group. CONCLUSIONS: We believe that FMS represents a new method in the conservative treatment of urinary incontinence. Magnetic stimulation with the Pulsegen device is efficient and safe. It can be used at home and, because of its small size, wearing the device is not annoying for patients. PMID- 12639648 TI - Preoperatively evaluated bladder wall tension as a prognostic parameter for postoperative success after surgery for bladder outlet obstruction. AB - OBJECTIVES: To evaluate a novel method for the determination of bladder wall tension (BWT) and to correlate these findings with postoperative persistent residual urine, postoperative uroflow, International Prostate Symptom Score, and quality-of-life index in patients with bladder outlet obstruction. METHODS: In 28 male patients with prostate enlargement or bladder neck sclerosis undergoing surgical treatment, the preoperative BWT was determined after urodynamic investigation and ultrasound determination of bladder weight. The patients were divided into two groups: group 1 (n = 24), postoperative residual urine volume less than 50 mL; and group 2 (n = 4), persistent residual urine volume greater than 50 mL. Five patients in group 1 were unobstructed in accordance with the Abrams-Griffiths nomogram. This group was compared separately with group 2, in which all 4 patients were also classified as unobstructed. RESULTS: The preoperative BWT in group 1 was 5.2 +/- 4.1 N/cm(2), significantly different from the preoperative BWT in group 2 (0.98 +/- 0.3 N/cm(2)). The BWT in the 5 unobstructed patients in group 1 (3.4 +/- 1.3 N/cm(2)) was significantly greater than that in patients in group 2. BWT was the sole parameter that was distinctly different between these 5 patients and the patients in group 2, with a significant influence on postoperative uroflow, International Prostate Symptom Score, and quality-of-life index. CONCLUSIONS: The results of this pilot study show that the determination of BWT allows further evaluation of the detrusor function. Especially in patients classified as unobstructed according to the Abrams-Griffiths nomogram, preoperative determination of the BWT could become a supplemental and important parameter with predictive value for postoperative success in patients with prostate enlargement or bladder neck sclerosis. PMID- 12639649 TI - Posterior tibial nerve stimulation in the treatment of idiopathic nonobstructive voiding dysfunction. AB - OBJECTIVES: To evaluate the effect of stimulation of the posterior tibial nerve in the treatment of voiding dysfunction. METHODS: Thirty-nine patients with chronic voiding dysfunction necessitating clean intermittent catheterization were enrolled in a prospective multicenter trial in the Netherlands (n = 19) and Italy (n = 20). They underwent 12 weekly sessions of posterior tibial nerve stimulation. Frequency/volume charts, an incontinence quality-of-life instrument, and the MOS 36-item Short-Form Health Survey were completed at 0 and 12 weeks. Subjective success was defined by the patient's positive response resulting in a request to continue treatment. Efficacy was based on analysis of the frequency/volume charts comparing the baseline values with the data at 12 weeks. A reduction of 50% or more in total catheterized volume was considered as an objective success (primary outcome measurement). RESULTS: Of the 39 patients, 23 (59%) chose to continue treatment. The frequency/volume charts showed a 50% decrease in total catheterized volume in 16 (41%) of 39 patients. Additionally, 10 patients (26%) noted a reduction of 25% to 50% in their total catheterized volume. For all patients, the total catheterized volume decreased by a mean of 228 mL (range -49 to -528). The incontinence quality-of-life instrument and Short Form Health Survey parameters improved significantly. CONCLUSIONS: Percutaneous stimulation of the posterior tibial nerve seems to be an effective, minimally invasive option worth trying in patients with idiopathic voiding dysfunction. Improvement was seen in objective micturition parameters, as well as in subjective quality-of-life data. PMID- 12639650 TI - Interstitial laser coagulation versus transurethral prostate resection for treating benign prostatic obstruction: a randomized trial with 2-year follow-up. AB - OBJECTIVES: To investigate whether interstitial laser coagulation (ILC) is as effective and as safe as transurethral resection of the prostate (TURP). The treatment of choice for bladder outflow obstruction secondary to benign prostatic hyperplasia is TURP. However, ILC is a less invasive outpatient procedure that may be as effective and safe as TURP. METHODS: In a multicenter randomized trial at six U.S. tertiary care hospitals, we treated 72 men with bladder outflow obstruction secondary to benign prostatic hyperplasia with either TURP (n = 35) or ILC (n = 37). The outcome measures were peak flow rate, postvoid residual urine volume, prostate volume, prostate-specific antigen levels, symptom and quality-of-life indexes, sexual function, and adverse event rates. Measurements were taken at baseline and at 3, 6, 12, and 24 months. RESULTS: At 2 years, the TURP patients had better median peak flow rates, but not significantly so (range 16.5 to 13.9 mL/s, 95% confidence interval for the 2.6 mL/s difference of -0.4 to 7.6). The median scores on the symptom indexes and quality-of-life measures were similarly improved in both groups. Of 37 ILC patients, 6 (16%) were retreated with TURP in the first year. Sexual function declined in the TURP group but remained stable in the ILC group. The adverse event rates were similar, although the events were more serious in the TURP group. CONCLUSIONS: ILC compares respectably with TURP. Given the advantages of an outpatient procedure, similar results in symptom reduction and quality-of-life measures, and less severe adverse effects, ILC can be an acceptable alternative to TURP. PMID- 12639651 TI - Incidence and severity of sexual adverse experiences in finasteride and placebo treated men with benign prostatic hyperplasia. AB - OBJECTIVES: To evaluate the incidence and resolution of sexual adverse experiences (AEs) in men with benign prostatic hyperplasia treated with finasteride 5 mg compared with placebo. METHODS: The Proscar Long-term Efficacy and Safety Study (PLESS) was a 4-year, randomized, double-blind, placebo controlled trial assessing the efficacy and safety of finasteride 5 mg in 3040 men, aged 45 to 78 years, with symptomatic benign prostatic hyperplasia, enlarged prostates, and no evidence of prostate cancer. Patients completed a questionnaire at screening regarding their history of sexual dysfunction. During treatment, spontaneously self-reported sexual AEs were recorded. RESULTS: At screening, 46% of patients in each treatment group reported some history of sexual dysfunction. During year 1 of the study, 15% of finasteride-treated patients and 7% of placebo treated patients had sexual AEs that were considered drug related by the investigator (P <0.001). During years 2 to 4, no between-group difference was noted in the incidence of new sexual AEs (7% in each group). The drug-related sexual AE profile for finasteride was similar for men with or without a history of sexual dysfunction. Sexual AEs resolved while continuing therapy in 12% of finasteride patients and 19% of placebo patients. Only 4% of finasteride and 2% of placebo patients discontinued the study because of sexual AEs. In men who discontinued with a sexual AE, 50% and 41% experienced resolution of their sexual AE after discontinuing finasteride or placebo therapy, respectively. CONCLUSIONS: Compared with placebo, men treated with finasteride experienced new drug-related sexual AEs with an increased incidence only during the first year of therapy. PMID- 12639652 TI - Periprostatic administration of local anesthesia during transrectal ultrasound guided biopsy of the prostate: a randomized, double-blind, placebo-controlled study. AB - OBJECTIVES: To evaluate the effect of periprostatic infiltration with local anesthesia on the level of discomfort associated with transrectal ultrasound guided needle biopsy of the prostate. Transrectal ultrasound-guided needle biopsy of the prostate is the standard procedure to diagnose prostate cancer. METHODS: A prospective, randomized, double-blind study was performed on 100 men referred for biopsy of the prostate. Fifty subjects were randomized to periprostatic injection of 10 mL of 2% lidocaine solution without epinephrine, and 50 were randomized to injection of placebo (10 mL of 0.9% NaCl). Each subject completed three 10-point visual analog scales for pain after a series of 15 needle biopsies. Pain was rated during the biopsy, diagnostic investigation, and injection of lidocaine. RESULTS: Patients who received local anesthesia had significantly lower visual analog scale scores compared with the group without lidocaine during the biopsy (mean score 0.76 versus 3.62, P <0.001) and diagnostic examination (mean score 1.08 versus 1.86, P = 0.025). Lidocaine injection caused no adverse effects. CONCLUSIONS: Periprostatic injection of lidocaine represents a simple and safe procedure that significantly reduces discomfort during probe manipulation and biopsy. We recommend this procedure in men undergoing transrectal ultrasound guided prostate biopsy. PMID- 12639653 TI - Using the percentage of biopsy cores positive for cancer, pretreatment PSA, and highest biopsy Gleason sum to predict pathologic stage after radical prostatectomy: the Center for Prostate Disease Research nomograms. AB - OBJECTIVES: To develop probability nomograms to predict pathologic outcome at the time of radical prostatectomy (RP) on the basis of established prognostic factors and prostate biopsy quantitative histology. METHODS: Using information from the database of the Center for Prostate Disease Research (CPDR), univariate and multivariate analyses were performed on 1510 men who had undergone transrectal ultrasound and biopsy for diagnosis and had radical prostatectomy as primary therapy, with variables of age, race, clinical stage, pretreatment prostate specific antigen (PSA), biopsy Gleason sum, and percentage of biopsy cores positive for cancer (total number of cores positive for cancer divided by the total number of cores obtained). The percentages of biopsy cores positive were grouped as less than 30%, 30% to 59%, and greater than or equal to 60%. The three most significant variables were used to develop probability nomograms for pathologic stage. RESULTS: PSA, biopsy Gleason sum, and percentage of cores positive were the three most significant independent predictors of pathologic stage. The assigned percentage of biopsy core-positive subgroups along with pretreatment PSA and highest Gleason sum were used to develop probability nomograms for pathologic stage. CONCLUSIONS: Pretreatment PSA, highest biopsy Gleason sum, and the percentage of cores positive for cancer are the most significant predictors for pathologic stage after radical prostatectomy. On the basis of these findings, CPDR probability nomograms were developed to predict pathologic outcome at the time of RP. PMID- 12639654 TI - Preoperative cardiopulmonary risk assessment as predictor of early noncancer and overall mortality after radical prostatectomy. AB - OBJECTIVES: To evaluate the capability of the preoperative cardiopulmonary risk assessment to predict early noncancer and overall mortality after radical prostatectomy for clinically localized prostate cancer. METHODS: In 444 consecutive radical prostatectomy patients, the American Society of Anesthesiologists Physical Status classification and the presence of cardiac insufficiency (New York Heart Association classification), angina pectoris (Canadian Cardiovascular Society classification), diabetes, hypertension, history of thromboembolism, and chronic obstructive or restrictive pulmonary disease were assessed. Kaplan-Meier time-event curves and Mantel-Haenszel hazard ratios were estimated for noncancer (other deaths were censored) and overall mortality. Cox proportional hazard models were used to analyze possible combined effects of risk factors. RESULTS: During an average follow-up of 4.7 years, 36 patients died: 15 of noncancer causes, 14 of prostate cancer, 6 of other cancers, and 1 in a car accident. The comorbidity scores for American Society of Anesthesiologists Physical Status classification, New York Heart Association classification, and Canadian Cardiovascular Society classification and combinations between the latter two scores were significantly associated with early noncancer mortality in a dose-response pattern. Furthermore, patients with chronic obstructive pulmonary disease were at increased risk. The association with overall mortality was less strong. CONCLUSIONS: The preoperative cardiopulmonary risk assessment may be used as a predictor of early noncancer and overall mortality after radical prostatectomy and should be evaluated further as a source of prognostic information in surgical oncology. PMID- 12639655 TI - Influence of body weight and prostate volume on intraoperative, perioperative, and postoperative outcomes after radical retropubic prostatectomy. AB - OBJECTIVES: To determine the influence of body weight and prostate volume on surgical outcomes after radical retropubic prostatectomy. METHODS: Between January 1994 and July 2000, 1024 men underwent radical retropubic prostatectomy by a single surgeon. Information was collected on body weight, prostate volume, operative time, estimated blood loss, allogenic transfusion rate, intraoperative complications, perioperative complications, postoperative complications, surgical margin status, and length of hospital stay. Postoperative continence and erectile function were assessed from self-administered questionnaires. RESULTS: Body weight was not significantly related to operative time (P = 0.09), estimated blood loss (P = 0.12), allogenic transfusion rate (P = 0.49), intraoperative complications (P = 0.37), perioperative complications (P = 0.84), postoperative complications (P = 0.44), positive margin rate (P = 0.07), length of hospital stay (P = 0.27), postoperative continence (P = 0.72), or postoperative erectile function (P = 0.92). The prostate volume was significantly and directly related to the estimated blood loss (P = 0.02), allogenic transfusion rate (P = 0.01), and length of hospital stay (P = 0.01). The prostate volume was significantly and inversely related to the positive surgical margin rate (P = 0.03). The prostate volume was not significantly related to the operative time (P = 0.12), intraoperative complications (P = 0.68), perioperative complications (P = 0.73), postoperative complications (P = 0.14), postoperative continence (P = 0.23), or postoperative erectile function (P = 0.90). CONCLUSIONS: No clinically relevant risks were associated with increasing body weight. Intraoperative bleeding resulting in higher transfusion rates represented the only clinically significant outcome that was adversely impacted by prostate volume. In the hands of experienced surgeons, radical retropubic prostatectomy can be performed in heavy men and in men with large prostates with excellent outcomes. PMID- 12639656 TI - Limited value of bone scintigraphy and computed tomography in assessing biochemical failure after radical prostatectomy. AB - OBJECTIVES: To define the utility of bone scan and computed tomography (CT) in the evaluation of patients with biochemical recurrence after radical prostatectomy. METHODS: A retrospective analysis of the Center for Prostate Disease Research database was undertaken to identify patients who underwent radical prostatectomy between 1989 and 1998. Patients who developed biochemical recurrence (two prostate-specific antigen [PSA] levels greater than 0.2 ng/mL) and underwent either bone scan or CT within 3 years of this recurrence were selected for analysis. The preoperative clinical parameters, pathologic findings, serum PSA levels, follow-up data, and radiographic results were reviewed. RESULTS: One hundred thirty-two patients with biochemical recurrence and a bone scan or CT scan were identified. Of the 127 bone scans, 12 (9.4%) were positive. The patients with true-positive bone scans had an average PSA at the time of the bone scan of 61.3 +/- 71.2 ng/mL (range 1.3 to 123). Their PSA velocities, calculated from the PSA levels determined immediately before the radiographic studies, averaged 22.1 +/- 24.7 ng/mL/mo (range 0.14 to 60.0). Only 2 patients with a positive bone scan had a PSA velocity of less than 0.5 ng/mL/mo. Of the 86 CT scans, 12 (14.0%) were positive. On logistic regression analysis, PSA and PSA velocity predicted the bone scan result (P <0.001 each) and PSA velocity predicted the CT scan result (P = 0.047). CONCLUSIONS: Patients with biochemical recurrence after radical prostatectomy have a low probability of a positive bone scan (9.4%) or a positive CT scan (14.0%) within 3 years of biochemical recurrence. Most patients with a positive bone scan have a high PSA level and a high PSA velocity (greater than 0.5 ng/mL/mo). PMID- 12639657 TI - Prospective comparison of short-term convalescence: laparoscopic radical prostatectomy versus open radical retropubic prostatectomy. AB - OBJECTIVES: To evaluate and compare prospectively the convalescence of patients after laparoscopic radical prostatectomy (LRP) and open radical retropubic prostatectomy (RRP) in a standardized clinical care pathway at a single institution by two surgeons of equal experience and training. METHODS: The study included all 60 patients undergoing LRP and RRP by two fellowship-trained surgeons in their first year of practice. The postoperative care of these patients was uniform and standardized. The medical records were reviewed and convalescence data obtained by an independent urologist and physician's assistant. RESULTS: Of the 60 patients, 24 underwent RRP and 36 underwent attempted LRP; 3 patients were converted from LRP to RRP. The differences in mean age, preoperative prostate-specific antigen level, Gleason score, in-hospital morphine equivalent requirement, time to oral intake, and hospital stay were not statistically significant between the LRP and RRP groups. The operating room time was significantly longer (5.8 +/- 1.2 hours versus 2.8 +/- 0.55 hours, P <0.0001) and the estimated blood loss was significantly lower in the LRP group (533 +/- 212 mL versus 1473 +/- 768 mL, P <0.0001) than in the RRP group. Pain medication use at home was significantly less in the LRP group (9 +/- 13 versus 17 +/- 15 oxycodone tablets, P <0.04), as was the time to complete convalescence (30 +/- 18 days versus 47 +/- 21 days, P <0.002). CONCLUSIONS: Although LRP took almost twice as long to complete as RRP in our initial clinical experience, the patients had a similar hospital course. LRP patients required less pain medication after discharge and had a shorter time to complete recovery than did RRP patients. Additional studies are needed to address long-term cancer control, potency, and continence outcomes to determine the precise role of LRP in the treatment of men with clinically localized prostate cancer. PMID- 12639658 TI - Assessment of surgical technique and perioperative morbidity associated with extraperitoneal versus transperitoneal laparoscopic radical prostatectomy. AB - OBJECTIVES: To compare the perioperative morbidity and surgical technique of the two main variants of laparoscopic radical prostatectomy: the more often used transperitoneal approach with primary dissection of the plane posterior to the seminal vesicles (TP-LRP) and a purely extraperitoneal approach (EP-LRP). METHODS: Our initial 20 consecutive patients who underwent EP-LRP and the last 20 consecutive patients who underwent TP-LRP were included in this retrospective study. The two groups were well matched for age, American Society of Anesthesiologists score, body mass index, prostate-specific antigen level, and Gleason score. The mean time for completion of the individual laparoscopic steps was assessed for both approaches. RESULTS: With EP-LRP, the mean operative time was shorter (169.6 minutes versus 224.2 minutes, P <0.001) and patients resumed a full diet earlier (mean 1.6 days versus 2.6 days, P = 0.002). The mean total dose and duration of morphine administration were higher in the TP-LRP group, but the difference was not statistically significant (6.0 mg versus 12.8 mg and 0.5 day versus 0.9 day, respectively). The catheter time was nearly identical in the two groups (mean 5.3 days with TP-LRP and 4.2 days with EP-LRP). The incidence of positive margins was not affected by the surgical approach. CONCLUSIONS: These preliminary results suggest that the initial posterior dissection associated with TP-LRP is less efficacious than direct access of the prevesical space during EP LRP. Measures of perioperative morbidity were at least equivalent, if not superior, for the EP-LRP technique. These findings, together with the significant decrease in operative time with EP-LRP, appear sufficiently important to abandon the transperitoneal technique. PMID- 12639659 TI - Neurophysiologic evaluation of central-peripheral sensory and motor pudendal pathways in primary premature ejaculation. AB - OBJECTIVES: Pudendal nerve somatosensory evoked potentials (SEPs), the bulbocavernosus (BC) reflex, and BC perineal motor evoked potentials after transcranial magnetic cortical stimulation were performed in patients with primary premature ejaculation to investigate the somatic sensory and motor function of the genital area. METHODS: Fourteen patients with primary premature ejaculation underwent psychological counseling, urologic physical examination, transrectal ultrasound examination, laboratory testing, and the Stamey test. The spinal and cortical pudendal nerve SEPs were performed by dorsal nerve stimulation at the penile shaft (DN-SEPs) in all patients and at the glans penis (GP-SEPs) in 3 of them. The BC reflex was obtained by stimulating the base of the penis. RESULTS: The mean sensory threshold did not significantly differ between the patients and normal subjects. Cortical DN-SEPs were normal in all patients. The sensory central conduction time, calculated in 6 patients, was normal. The mean cortical DN-SEP amplitude was significantly smaller in patients than in controls. In 3 patients and in 3 controls who underwent both DN-SEP and GP-SEP testing, the glans penis sensory threshold was lower than the dorsal nerve threshold and the cortical GP-SEP latency was longer than the cortical DN-SEP latency. The BC reflex was normal in most patients. The BC motor evoked potentials were normal in all patients, but one. CONCLUSIONS: We did not confirm either a faster conduction along the pudendal sensory pathway or a greater cortical representation of the sensory stimuli from the genital area in our patients. Moreover, we did not confirm hyperexcitability of the BC reflex in them. Our results suggest that the electrophysiologic approach is probably not sufficient to clarify the causes of primary premature ejaculation. A more integrated investigation could allow better results in this field. PMID- 12639660 TI - Serum androgen levels in men: impact of health status and age. AB - OBJECTIVES: To investigate the impact of health status on androgen levels in men and to assess the age-related changes in androgen levels. METHODS: Sera were obtained from 526 men (age 20 to 89 years) participating in a health screening project and 35 men selected according to the SENIEUR protocol, which excludes those with underlying disease. The health screening project participants were divided into two groups: healthy men (n = 133; group 1) and the remainder (n = 393; group 2). The total testosterone and sex hormone-binding globulin levels were quantified, and the amount of free testosterone (cfT) was calculated. RESULTS: The lowest annual declines in androgen level were observed in group 3 (testosterone 0.2%; cfT 0.4%), followed by group 1 (testosterone 0.4%; cfT 0.8%) and group 2 (testosterone 0.8%; cfT 0.9%). Body mass index and levels of cholesterol, triglycerides, and glucose correlated negatively (P <0.01) with testosterone and cfT. Decade-adjusted reference values (mean +/- 2 standard deviations) for testosterone and cfT were defined. The proposed lower testosterone reference value declined gradually from 3.1 ng/mL (20 to 29 years) to 1.7 ng/mL (70 years or older). CONCLUSIONS: In this cross-sectional study, serum androgens declined in aging men, and the extent of this decrease correlated with health status. A fixed, non-age-adjusted reference value does not adequately reflect this and might lead to a false diagnosis of androgen deficiency, particularly in elderly men. The clinical value of age-related reference values, however, needs to be determined in prospective studies. PMID- 12639661 TI - Management of mullerian duct remnants. AB - OBJECTIVES: To describe the presentation and radiologic investigation of 6 patients who had persistent mullerian duct remnants (MDRs) in association with severe hypospadias and to review the long-term results of surgical management using the transtrigonal approach. METHODS: The hospital records of 6 patients who had MDRs and who underwent transvesical transtrigonal excision were reviewed and long-term assessment of urinary control and continence was made by interview. RESULTS: One patient presented in the newborn period with clinically palpable MDRs and five others escaped detection until after the hypospadias repair, despite ultrasonography of the pelvis specifically looking for MDRs. All had undescended testis with severe hypospadias. The karyotype was 45XO/46XY in five and 46XY in one. The most consistently useful examinations were voiding cystourethrography and magnetic resonance imaging. The excision was transtrigonal in each case. Early or late postoperative complications were not encountered in any of our patients. Gonadal histologic examination showed streak gonad in four and a dysgenetic testis in one. Histopathologic examination showed rudimentary uterus and fallopian tubes in 4 patients and a cystic structure lined with squamous and columnar epithelium in 2 patients. No areas of metaplasia were noted. Bladder emptying was normal after surgery. CONCLUSIONS: Ultrasound examination alone is not always sufficient to diagnose MDRs, which may be responsible for symptoms after hypospadias surgery. The transtrigonal approach provided excellent visualization and access to MDRs and prostatic urethra. PMID- 12639665 TI - Metastatic knee. PMID- 12639663 TI - Testicular torsion: is there a seasonal predilection for occurrence? AB - OBJECTIVES: To test the hypothesis that there is a bimodal spring-fall seasonal predilection for acute testicular torsion and to compare our series with those previously reported outside the United States that support a seasonal predilection. METHODS: A retrospective chart review of operative notes was performed for all cases of acute torsion of the testis confirmed at surgery during a 15-year period at our institution. The diagnosis, child's age, and date of occurrence were verified; 135 confirmed cases were identified. RESULTS: The months with the highest incidences of acute testicular torsion were December (14%) and January (11%). Thirty percent of cases occurred in the fall (October, November, December), 24% in the winter (January, February, March), 22% in the summer (July, August, September), and 23% in the spring (April, May, June). CONCLUSIONS: No statistically significant differences were seen with regard to seasonal or monthly occurrence of acute torsion. However, the "cold" weather trend identified is consistent with those of larger international studies. It is possible that our regional monthly temperatures do not vary enough to demonstrate whether cold weather influences the incidence of torsion. To our knowledge, this is the only reported series regarding a seasonal predilection for torsion in the United States. A multi-institutional, prospective study representing all regions within the United States is needed to establish the significance of season and climate on testicular torsion. PMID- 12639666 TI - Donor to native ureteroureterostomy in a patient with pre-existing ileal conduit during renal transplantation. PMID- 12639667 TI - Pinball calculus in a ureterosigmoidostomy. AB - We report a complication during the treatment of lithiasis with extracorporeal shock wave lithotripsy in a patient with a ureterosigmoidostomy. This woman presented with renal colic bilaterally and renal insufficiency and was found to have an extremely mobile calculus. A significant gaseous reflux from the sigmoid colon was found to propel the solitary calculus in a retrograde fashion across the ureteroileal anastomosis up the ureter into one kidney, and then later, after re-descent to the level of the anastomosis, up into the opposite kidney. After several days of playing hide and seek with this migrating calculus, using extracorporeal shock wave lithotripsy, the patient became stone free. PMID- 12639668 TI - Intravesical phallus in cloacal exstrophy. AB - We report a case of a fused phallus located within the urinary bladder in a child with cloacal exstrophy. Surgical exploration revealed a phallus formed by fusion of the left and right corpus cavernosum and completely covered by bladder urothelium. The phallus was surgically separated from the bladder and mobilized to a more normal position and a neourethra created from tubularized bladder urothelium. Skin coverage was done using perineal skin flaps to cover the penile shaft, with the bladder urothelium covering the tip of the phallus being left intact to provide the appearance of a glans penis. PMID- 12639669 TI - Pediatric pyogenic granuloma of the glans penis. AB - Pyogenic granulomas are benign vascular proliferations of the skin and mucous membranes. We present a case report of a 13-year-old uncircumcised boy with phimosis and a pyogenic granuloma of the glans penis. The relationship between these lesions, phimosis, smegma, and circumcision is discussed. When the lesion is found in conjunction with phimosis, consideration should be given for circumcision. Close follow-up to rule out recurrence is necessary. PMID- 12639670 TI - Clear cell adenocarcinoma in a female urethral diverticulum. AB - Primary urethral carcinoma is an uncommon diagnosis, and carcinomas arising from within a urethral diverticulum are rare. Because of the limited number of diagnosed cases, optimal treatment guidelines are not available. However, patients require an aggressive treatment approach to provide the best chance for cure. PMID- 12639671 TI - Complete resection of synovial sarcoma of prostatic fascia. AB - Synovial sarcomas primarily occur chiefly in the para-articular regions of the extremities and extremely rarely in the prostatic region. We report a patient with synovial sarcoma of the prostatic fascia. The patient underwent retropubic radical prostatectomy and at 6 months of follow-up, no evidence of another lesion or local recurrence was found. PMID- 12639672 TI - Durability of the Malone antegrade continence enema in pregnancy. AB - The Malone antegrade continence enema (MACE) is a therapeutic option to treat chronic constipation and fecal incontinence in patients with neurogenic bowel. Previous reports have described the short-term success of this procedure, but no report has described the durability of the procedure during pregnancy. We present the case of a spinal cord injury patient who underwent an uncomplicated pregnancy after a MACE procedure with no stomal catheterization difficulties or leakage during or after the pregnancy. PMID- 12639673 TI - Multilocular prostatic cystadenoma with high-grade prostatic intraepithelial neoplasia. AB - Multilocular prostatic cystadenoma is a rarely encountered neoplasm located in the midline between the bladder and rectum that is either attached to the prostate by a pedicle or separate from the prostate entirely. Histologically and immunohistochemically these lesions resemble benign prostate tissue. We report the first case of this entity for which multifocal high-grade prostatic intraepithelial neoplasia (PIN) is identified. Conceptually, the finding of high grade PIN in multilocular prostatic cystadenomas provides further evidence that these lesions are fully analogous to the prostate gland not only in their morphology and immunohistochemistry but also in their predilection for the same diseases. PMID- 12639674 TI - Successful treatment of arterial priapism with radiologic selective transcatheter embolization of the internal pudendal artery. AB - We report our successful treatment of arterial priapism by means of radiologic selective transcatheter embolization of the internal pudendal artery using an absorbable gelatin sponge (Spongel). PMID- 12639675 TI - Spontaneous expulsion of a screw during urination: an unusual complication 9 years after internal fixation of pubic symphysis diastasis. AB - Nine years after treatment of symphysiolysis and dislocation of the left sacroiliac joint, a screw was spontaneously voided during urination. Unstable plate fixation of the symphysis pubis probably caused screw migration into the bladder, creating a fistula with abscess formation and septic complications. PMID- 12639676 TI - Pulsed radiofrequency as a treatment for groin pain and orchialgia. AB - Inguinal and testicular pain are challenging problems for which no reliable, standardized treatment exists. We report 3 patients with groin pain or orchialgia who were treated with pulsed radiofrequency of the nerves innervating these areas. All 3 patients reported complete pain relief at their 6-month follow-up visits. The techniques and settings used for the nerve blocks and radiofrequency procedures are explained in detail, along with a brief synopsis of the rationale for using it. Randomized, placebo-controlled studies are needed to better assess the efficacy of this procedure and identify eligible candidates. PMID- 12639677 TI - Role of mitochondrial cytochrome c in cocaine-induced apoptosis in rat testes. AB - OBJECTIVES: We have previously demonstrated that cocaine exposure leads to apoptosis in rat testes. To understand further the mechanism of cocaine-induced testicular damage, we studied the effect of cocaine on cytochrome c release from the mitochondria. We also determined the caspase 3, caspase 8, and caspase 9 activities in rat testes after chronic cocaine exposure. METHODS: Thirty-day-old male Sprague-Dawley rats received cocaine hydrochloride or equal volumes of normal saline subcutaneously daily for 90 days. The testes were removed at 15, 30, and 90 days of cocaine or saline administration. Mitochondria and cytosolic fractions from testes were isolated. Western blotting was performed in both fractions using anti-cytochrome c antibody. Caspase 3, caspase 8, and caspase 9 activities were determined by fluorometric assay. RESULTS: The expression of cytochrome c protein in the cytosolic fraction was increased on day 15 and persisted for up to 90 days after cocaine injection compared with controls. However, the expression of cytochrome c in testes was decreased in the mitochondria fraction on days 15, 30, and 90 after cocaine injections compared with the corresponding controls. The caspase activity study showed caspase 3 and caspase 9 activities increased in cocaine-treated testes at each point of the study compared with the corresponding controls. However, the caspase 8 activity in cocaine-treated testes did not change significantly at each point of the study compared with the corresponding controls. CONCLUSIONS: Our results suggest that the release of cytochrome c from mitochondria and its subsequent activation of caspase 9 and caspase 3 in testes play a key role in cocaine-induced germ cell apoptosis. Our findings also indicate that cocaine-induced testicular germ cell apoptosis in rats is at least initiated through a mitochondria-associated pathway. PMID- 12639678 TI - Effect of castration on acetyl salicylic acid metabolism in rabbits. AB - OBJECTIVES: The biotransformation of acetyl salicylic acid (ASA) differs within species, and gender differences have been documented and attributed to the effect of sex hormones. Castration remains a standard therapy for men with advanced prostate cancer. We studied the effect of castration on the metabolism of ASA in rabbits to find out whether the metabolism of ASA is adversely affected after castration. METHODS: ASA in doses of 12.5, 25, and 50 mg/kg body weight was given intravenously to male and female prepubertal and adult rabbits, castrated adult male rabbits, and castrated male rabbits given testosterone (3 animals per group). Blood samples were collected at 0, 10, 30, 60, 120, and 180 minutes. The high-performance liquid chromatography method was used for the quantitation of salicylic acid (SA) in serum. The percentage of SA not metabolized was determined by comparing the serum level at 10 and 180 minutes for each group. RESULTS: At a dose of 50 mg/kg in the adult rabbits, the mean +/- SD of SA in serum at 10 and 180 minutes was 146.54 +/- 29.54 microg/mL and 19.12 +/- 5.93 microg/mL for males, 158.25 +/- 6.70 microg/mL and 33.24 +/- 2.78 microg/mL for females, 229.72 +/- 47.85 microg/mL and 44.33 +/- 5.64 microg/mL for castrated male rabbits, and 170.88 +/- 12.03 microg/mL and 68.1 +/- 37.54 microg/mL for castrated male rabbits given testosterone, respectively. Also, at 180 minutes, the percentage of SA not metabolized in adult male rabbits was 12.82% +/- 1.65% compared with 21.04% +/- 2.14% (P <0.01) in adult females, 19.53% +/- 1.73% (P <0.01) in castrated adult male rabbits, and 38.95% +/- 19.48% (P <0.001) in castrated male rabbits given testosterone. At all doses of ASA, the serum SA concentration in male and female prepubertal rabbits was not significantly different for each time point. CONCLUSIONS: These results indicate that male rabbits are able to metabolize ASA faster than are females. After castration, this ability is significantly decreased. If these experimental results are confirmed in humans, men who are undergoing hormonal manipulation for advanced prostate cancer and who require high-dose ASA, such as in the treatment of stroke or rheumatoid arthritis or as an antioxidant, may need lower doses to reduce the possible toxic effects of ASA. PMID- 12639679 TI - Intravesical liposome administration--a novel treatment for hyperactive bladder in the rat. AB - OBJECTIVES: To examine the effect of intravesical administration of liposomes (LPs) on chemically induced bladder hyperactivity in the rat. It has been suggested that interstitial cystitis (IC) is associated with a dysfunctional or leaky epithelium. Thus, enhancement of epithelial barrier function might be useful in the treatment of IC. LPs are vesicles that are concentric phospholipid bilayers separated by an aqueous compartment and can fuse with cells to provide a molecular film that can promote wound healing. METHODS: The intravesical pressure was recorded using a transurethral catheter in adult female Sprague-Dawley rats anesthetized with urethane (1.2 g/kg subcutaneously). Some animals were pretreated with capsaicin (125 mg/kg subcutaneously) 4 days before the experiments. Continuous cystometrograms were performed by slowly filling the bladder (0.04 mL/min) with solutions of varying compositions, including saline, acetic acid (AA, 0.1%), potassium chloride (KCl, 500 mM), protamine sulfate (PS, 10 mg/mL), LPs, PS/KCl, or LPs/KCl. The parameters measured included the intercontraction interval (ICI), amplitude of bladder contractions, compliance, and micturition pressure threshold. RESULTS: The ICI was decreased after exposure to AA (79.8% decrease) or PS/KCl (81% decrease); however, the ICI was not changed after LPs, PS, or KCl alone. The decreased ICI was partially reversed after infusion of LPs (172.8% increase) or LPs/KCl (63% increase), but was not significantly changed after switching to saline or KCl administration. Pretreatment with capsaicin delayed the onset of the irritative effects of AA by approximately 30 to 60 minutes, but had not changed the magnitude after 2 hours of infusion. CONCLUSIONS: Intravesical administration of PS/KCl or AA activates capsaicin-sensitive and capsaicin-resistant afferents in a time-dependent sequence that is partially reversed by LP infusion. We hypothesize that LPs might enhance the barrier properties of a dysfunctional uroepithelium and increase resistance to irritant penetration. Thus, intravesical LP administration could be a novel treatment of patients with IC. PMID- 12639681 TI - Effects of methoctramine on bladder overactivity in a rat model. AB - OBJECTIVES: To determine the functional effects of methoctramine as an M(2) muscarinic receptor antagonist on isolated detrusor strips in vitro and bladder overactivity in vivo in rats. METHODS: A total of 114 Sprague-Dawley rats were used in the present study. Isolated rat detrusor strips were contracted by depolarizing the preparations with carbachol. Methoctramine was added to the tissue bath in increasing concentrations, and contraction inhibition was assessed. Isovolumetric contractions were evoked by electrical stimulation using a bipolar electrode. Efficacy against bladder instability was evaluated using the obstructed hypertrophied bladder model in the rat. The acetic acid bladder cystometry model was used to assess the efficacy of methoctramine in neurogenic detrusor overactivity. RESULTS: Methoctramine inhibited carbachol-induced bladder contractions significantly in isolated rat detrusor strips in a concentration dependent manner. The amplitude of electrically evoked isovolumetric contractions was decreased significantly after methoctramine exposure. In vivo methoctramine administered intravenously significantly increased the voiding interval and bladder compliance. In addition, a decrease occurred in the number of spontaneous contractions during the filling phase in a model of neurogenic and obstruction induced detrusor overactivity. CONCLUSIONS: M(2) antagonists in general may represent a new useful class of drug worth considering in the treatment of bladder overactivity. PMID- 12639680 TI - Intravesical protamine sulfate and potassium chloride as a model for bladder hyperactivity. AB - OBJECTIVES: An acute animal model for hyperactive bladder in rats was developed using intravesical infusion of protamine sulfate (PS), an agent thought to break down urothelial barrier function, and physiologic concentrations of potassium chloride (KCl). METHODS: Continuous cystometrograms (CMGs) were performed in urethane-anesthetized female rats by filling the bladder (0.04 mL/min) with normal saline followed by intravesical infusion of a test solution consisting of either KCl (100 or 500 mM) or PS (10 or 30 mg/mL) for 60 minutes. Subsequently, the 10 mg/mL PS-treated animals were infused intravesically with 100, 300, or 500 mM KCl. Some animals were pretreated with capsaicin (125 mg/kg subcutaneously) 4 days before the experiments. RESULTS: Unlike KCl (100 or 500 mM) or a low concentration of PS (10 mg/mL) alone, the intravesical administration of a high concentration of PS (30 mg/mL) produced irritative effects with a decreased intercontraction interval (by 80.6%). After infusion of a low concentration of PS, infusion of 300 or 500 mM KCl produced irritative effects (intercontraction interval decreased by 76.9% or 82.9%, respectively). The onset of irritation occurred more rapidly after 500 mM KCl (10 to 15 minutes) than after 300 mM KCl (20 to 30 minutes). Capsaicin pretreatment delayed the onset (approximately 60 minutes) and reduced the magnitude (intercontraction interval decreased by 35.5%) of irritative effects. CONCLUSIONS: Intravesical administration of KCl after PS treatment activates capsaicin-sensitive afferents and detrusor muscle and presumably capsaicin-resistant afferents. Modest, noncytotoxic affronts to urothelial barrier function can result in dramatic irritative responses. This model may be useful in the study of bladder irritation and hyperactivity. PMID- 12639683 TI - New dimensions in color perception. AB - Colors are generally ordered in three dimensions, with hue and saturation as polar coordinates of a color circle, and brightness as the third dimension. Intuitively, lines of constant hue (but variable saturation) in such a color space should converge on an achromatic point devoid of hue. However, in new experiments by Ekroll et al. using colored patches in colored surrounds, constant hue lines converge not on 'gray' but on the surround color. This paradoxical observation suggests that the standard three-dimensional conception of perceived color is inadequate. PMID- 12639682 TI - Clinical significance of heat shock protein-70 expression in bladder cancer. AB - OBJECTIVES: To investigate the expression of heat shock protein-70 (HSP-70) in transitional cell bladder cancer and correlate it with normal and inflammatory bladder tissue samples. METHODS: We used a three-step avidin-biotin method to examine the localization of a rabbit anti-HSP-70 polyclonal antibody in normal (n = 10), inflammatory (n = 10), and malignant (n = 67) bladder tissue samples. RESULTS: Fifty-eight percent of the bladder cancer samples and only 10% and 20% of the normal and inflammatory bladder tissue samples, respectively, expressed HSP-70. Localization of the antibody correlated with the grade (P <0.001), stage (P <0.001), and survival (P <0.05) of the patients with malignancy. CONCLUSIONS: HSP-70 is frequently overexpressed by bladder cancer cells and could be used as biochemical marker in patients with bladder cancer. PMID- 12639684 TI - Taking the MAX from neuronal responses. AB - By taking the MAX from their inputs, neurons in the ventral visual pathway might preserve their selectivity even when stimulated with natural scenes. This computational hypothesis has received recent direct physiological evidence from recordings of V4 neuronal responses, in a recent study by Gawne and Martin (2002). Object vision might rely more heavily on parallel processing than generally thought. PMID- 12639685 TI - The nature of cognitive development. AB - Theories of cognitive development have led to enduring and fierce arguments that have been long on rhetoric but short on evidence. Constructivist theory has roots in Piagetian notions of cognitive development as proceeding from self-directed action during infancy. Nativist theories subsequently became popular by producing claims of cognitive precocity, but left open many central questions concerning mechanisms of development. Now, a new view of constructivism is experiencing a renaissance, having achieved greater psychophysical, computational and neural plausibility. PMID- 12639686 TI - Out-of-body experiences: from Penfield to present. AB - Can the brain, when stimulated, yield entirely novel experiences? Blanke et al. (2002) describe a patient who reported spontaneous out-of-body experiences during electrical stimulation of her angular gyrus. These findings, although apparently extraordinary, agree with much earlier reports from a patient tested by Wilder Penfield. Such studies can provide clues about the nature of conscious experience. PMID- 12639687 TI - Show us the model. PMID- 12639688 TI - The past-tense debate: exocentric form versus the evidence. PMID- 12639689 TI - Beyond one model per phenomenon. PMID- 12639690 TI - Mental imagery: against the nihilistic hypothesis. PMID- 12639691 TI - Explaining mental imagery: now you see it, now you don't. Reply to Kosslyn et al. PMID- 12639692 TI - Return of the mental image: are there really pictures in the brain? AB - In the past decade there has been renewed interest in the study of mental imagery. Emboldened by new findings from neuroscience, many people have revived the idea that mental imagery involves a special format of thought, one that is pictorial in nature. But the evidence and the arguments that exposed deep conceptual and empirical problems in the picture theory over the past 300 years have not gone away. I argue that the new evidence from neural imaging and clinical neuropsychology does little to justify this recidivism because it does not address the format of mental images. I also discuss some reasons why the picture theory is so resistant to counterarguments and suggest ways in which non pictorial theories might account for the apparent spatial nature of images. PMID- 12639693 TI - Religious thought and behaviour as by-products of brain function. AB - Religious concepts activate various functionally distinct mental systems, present also in non-religious contexts, and 'tweak' the usual inferences of these systems. They deal with detection and representation of animacy and agency, social exchange, moral intuitions, precaution against natural hazards and understanding of misfortune. Each of these activates distinct neural resources or families of networks. What makes notions of supernatural agency intuitively plausible? This article reviews evidence suggesting that it is the joint, coordinated activation of these diverse systems, a supposition that opens up the prospect of a cognitive neuroscience of religious beliefs. PMID- 12639694 TI - Spatial cognition: evidence from visual neglect. AB - Recent work on human attention and representational systems has benefited from a growing interplay between research on normal attention and neuropsychological disorders such as visual neglect. Research over the past 30 years has convincingly shown that, far from being a unitary condition, neglect is a protean disorder whose symptoms can selectively affect different sensory modalities, cognitive processes, spatial domains and coordinate systems. These clinical findings, together with those of functional neuroimaging, have increased knowledge about the anatomical and functional architecture of normal subsystems involved in spatial cognition. We provide a selective overview of how recent investigations of visual neglect are beginning to elucidate the underlying structure of spatial processes and mental representations. PMID- 12639695 TI - Task switching. AB - Everyday life requires frequent shifts between cognitive tasks. Research reviewed in this article probes the control processes that reconfigure mental resources for a change of task by requiring subjects to switch frequently among a small set of simple tasks. Subjects' responses are substantially slower and, usually, more error-prone immediately after a task switch. This 'switch cost' is reduced, but not eliminated, by an opportunity for preparation. It seems to result from both transient and long-term carry-over of 'task-set' activation and inhibition as well as time consumed by task-set reconfiguration processes. Neuroimaging studies of task switching have revealed extra activation in numerous brain regions when subjects prepare to change tasks and when they perform a changed task, but we cannot yet separate 'controlling' from 'controlled' regions. PMID- 12639696 TI - The cognitive revolution: a historical perspective. AB - Cognitive science is a child of the 1950s, the product of a time when psychology, anthropology and linguistics were redefining themselves and computer science and neuroscience as disciplines were coming into existence. Psychology could not participate in the cognitive revolution until it had freed itself from behaviorism, thus restoring cognition to scientific respectability. By then, it was becoming clear in several disciplines that the solution to some of their problems depended crucially on solving problems traditionally allocated to other disciplines. Collaboration was called for: this is a personal account of how it came about. PMID- 12639697 TI - New approaches to treatment of various cancers based on cytotoxic analogs of LHRH, somatostatin and bombesin. AB - The development of targeted cytotoxic analogs of hypothalamic peptides for the therapy of various cancers is reviewed and various oncological studies on experimental tumors are summarized. Novel therapeutic modalities for breast, prostate and ovarian cancer consist of the use of targeted cytotoxic analogs of LH-RH containing doxorubicin (DOX) or 2-pyrrolino-DOX. The same radicals have been incorporated into cytotoxic analogs of somatostatin which can be also targeted to receptors for this peptide in prostatic, mammary, ovarian, renal and lung cancers, brain tumors and their metastases. A targeted cytotoxic analog of bombesin containing 2-pyrrolino-DOX has also been synthesized and successfully tried in experimental models of prostate cancer, small cell lung carcinoma and brain tumors. The development of these new classes of peptide analogs should lead to a more effective treatment for various cancers. PMID- 12639698 TI - Acute effects of troglitazone and nitric oxide on glucose uptake in L929 fibroblast cells. AB - The thiazolidinedione class of antidiabetic drugs, including troglitazone, has an insulin-sensitizing effect for patients with type 2 diabetes. However, in some tissues, studies have shown that troglitazone also has an acute insulin independent effect on glucose uptake. To determine the extent of this acute action of troglitazone, the effect of troglitazone on 2-deoxyglucose (2DG) uptake in L929 fibroblast cells was measured. Troglitazone stimulated 2DG uptake in a dose dependent manner with a maximum stimulation of >300% at 5-10 microM. In addition, nitric oxide has been shown to stimulate glucose uptake in peripheral muscle tissue. Therefore, the effect of nitric oxide on 2DG uptake in L929 cells was also investigated using the nitric oxide donor, sodium nitroprusside (SNP). SNP stimulated 2DG uptake by >200% with a maximally effective concentration of 5 mM. The combined effect of maximally effective concentrations of both stimulants (10 microM troglitazone + 5 mM SNP) was not additive suggesting a shared pathway for 2DG uptake. However, the nitric oxide synthase inhibitor, N(G)-monomethyl-L arginine (L-NMMA, 50 microM) had no effect on troglitazone stimulated 2DG uptake, indicating that the troglitazone and nitric oxide pathways converge after nitric oxide production. In addition, 12.5 microM dantrolene was shown to have no effect on either troglitazone or SNP stimulated 2DG uptake suggesting that these stimulatory effects are independent of changes in calcium ion concentrations. These data provide important evidence for the acute regulation of glucose transport through GLUT 1 transporters. PMID- 12639699 TI - Proerectile effects of apomorphine in mice. AB - Dopaminergic pathways play a key role in the central control of sexual behavior. Stimulation of central dopaminergic receptors elicits penile erection in a variety of species and has been proposed as a treatment option for erectile dysfunction in humans. The present study investigated the proerectile effects of apomorphine in mice. In this species, subcutaneous injection of apomorphine (range: 0.11-110 microg/kg sc) elicited three different behavioral responses: erection, erection-like responses and genital grooming. Proerectile effects of apomorphine were dose-dependent. More than 50% of mice displayed erections after administration of 1.1-11 microg/kg of apomorphine sc. Proerectile effects of apomorphine were blocked by haloperidol, a central D2 antagonist, but not by domperidone, a peripherally active dopaminergic antagonist. We conclude that apomorphine elicits erection in mice. This effect is dose-dependent and due to activation of central D2 dopaminergic receptors. The mouse model may be useful for pharmacological approaches designed to provide a better understanding of the central mechanisms of penile erection and sexual behavior. PMID- 12639700 TI - Interaction of tocopherols and phenolic compounds with membrane lipid components: evaluation of their antioxidant activity in a liposomal model system. AB - This paper describes the use of complex liposomes as real membrane models to evaluate the potential benefits of several antioxidants in relation to lipid peroxidation. The xanthine oxidase/Fe(3+)-ADP-EDTA and the Fe(2+)/H2O2 systems have been used to generate hydroxyl radicals and the water soluble azo-compound 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) to generate carbon centered radicals (A*) by thermal decomposition. The antioxidant behavior of the rosemary and citrus plant extracts and vitamin-E and vitamin-E acetate alpha-tocopherols have been analyzed. The order of effectiveness in avoiding radical chain reactions has been established by using the colorimetric thiobarbituric acid reaction and the fluorescent probe DPH-PA. ESR spectroscopy has been used to carry out the pursuit of the oxidation processes on the basis of the identification of the radical species resulting from the oxidant system and the ability of the antioxidants to act as scavengers for hydroxyl and AAPH-derived radicals. The modification of the main transition temperature for the lipid mixture and the splitting of the calorimetric peak in the presence of the antioxidants were demonstrated by differential scanning calorimetry. The results obtained showed that the phenols-containing plant extracts and alpha-tocopherols perturb the phase behavior of the BBE lipid bilayer and have a fluidifying effect that could favor the known antioxidant capability and scavenging characteristics of these compounds. 31P-NMR results could be interpreted as, after the incorporation of these antioxidants, those lipid molecules interacting with antioxidants give rise to lamellar phase spectral components with resonance position at lower fields or to isotropic signals in accordance with a higher motion of their phosphate groups. PMID- 12639701 TI - Reduction of inflammation in rats by diazepam: tolerance development. AB - High doses of diazepam (10-20 mg/kg) were shown to reduce the volume of acute carrageenan-induced inflammatory paw edema in rats. This effect was not observed after adrenalectomy or long-term use of similar doses of diazepam. The present experiment was undertaken to analyze the effects of long-term (21 daily injections) treatment with diazepam (10 mg/kg) on both carrageenan-induced paw edema (CIPE) and corticosterone serum levels. For comparison, the effects of a single and acute 10 mg/kg dose of diazepam were also analyzed. Results showed that: 1- long-term diazepam treatment induced no changes in CIPE values and corticosterone serum levels; 2- acute diazepam treatment reduced CIPE values and increased corticosterone serum levels; 3- the plasmatic levels of diazepam measured 1 hour after the single treatment or 1 hour after the last dose of long term diazepam administration were not different. These results indicate the development of tolerance to diazepam effects on both CIPE and corticosterone serum levels and suggest a relevant role for corticosterone on diazepam-induced inhibition of acute inflammation. Data were discussed in the light of peripheral benzodiazepine receptor site (PBR) activation within adrenal gland cells by diazepam, thereby increasing the serum levels of corticosterone and thus reducing CIPE. Possible actions of diazepam on HPA axis activity and/or on cytokine network were also discussed. PMID- 12639702 TI - Red wine micronutrients as protective agents in Alzheimer-like induced insult. AB - Plant extract micronutrients are commonly added to diets for health and prevention of degenerative disease. However, there are barriers to the introduction of these products as antioxidant therapies in counteracting chronic human diseases, probably because the molecular bases of their therapeutic potential are poorly clarified. The present study was designed to evaluate the possible protective effect of combined micronutrients present in black grape skin on toxicity induced by 25-35 beta-amyloid peptid or by serum of Alzheimer's disease patients, in human umbilical vein endothelial cells (HUVECs). The hypothesis was tested by examining the results of lactic dehydrogenase (LDH) release to estimate cytoplasmic membrane breakdown; activity of mitochondrial complexes, reactive oxygen species (ROS) production and malonyl dialdehyde (MDA) levels as markers of oxidative stress induction and COMET assay to evaluate DNA fragmentation. The results demonstrate that black grape skin extract reduces the ROS production, protects the cellular membrane from oxidative damage, and consequently prevents DNA fragmentation. The experimental results suggest that this natural compound may be used to ameliorate the progression of pathology in AD disease therapy. PMID- 12639703 TI - Triterpene-enriched extracts from Ganoderma lucidum inhibit growth of hepatoma cells via suppressing protein kinase C, activating mitogen-activated protein kinases and G2-phase cell cycle arrest. AB - The medicinal mushroom Ganoderma lucidum (G. lucidum) has been used in the Orient for the prevention and treatment of various diseases including cancer. Except for the immune enhancing properties of its polysaccharide constituent, very little is known about the anticancer activity of another major constituent, triterpenes. In this report, we studied the anticancer mechanism of triterpene-enriched extracts from G. lucidum. The triterpene-enriched fraction, WEES-G6, was prepared from mycelia of G. lucidum by sequential hot water extraction, removal of ethanol insoluble polysaccharides and then gel-filtration chromatography. We found that WEES-G6 inhibited growth of human hepatoma Huh-7 cells, but not Chang liver cells, a normal human liver cell line. Treatment with WEES-G6 caused a rapid decrease in the activity of cell growth regulative protein, PKC, and the activation of JNK and p38 MAP kinases. The changes in these molecules resulted in a prolonged G2 cell cycle phase and strong growth inhibition. None of these effects were seen in the normal liver cells. Our findings suggest that the triterpenes contained in G. lucidum are potential anticancer agents. PMID- 12639704 TI - Nucleotide changes in the translated region of SCN5A from Japanese patients with Brugada syndrome and control subjects. AB - The mutations of the SCN5A gene have been implicated to play a pathogenetic role in Brugada syndrome, which causes ventricular fibrillation. To determine the Brugada-associated mutations in Japanese patients, facilitate pre-symptomatic diagnosis, and allow genotype-phenotype studies, we screened unrelated patients with Brugada syndrome for mutations. DNAs from 6 Japanese patients were obtained and the sequence in the translated region of SCN5A was determined. We could not find the mutations reported previously, but found 17 sites of nucleotide change, consisting of 7 synonymous and 10 non-synonymous nucleotide changes in our patients. Among them, two non-synonymous nucleotide changes (G1663A and G5227A) are specific to our patients and these changes were not found in 53 healthy controls. In 4 patients out of 6, no specific nucleotide change for Brugada syndrome could be detected. Our findings demonstrating no patient-specific change in the translated region of the SCN5A gene among two thirds of the small number of patients examined here imply that another gene other than the SCN5A may be associated with this disease, supporting previous investigations in Japan and other countries. PMID- 12639705 TI - Age and anesthetic effects on murine electrocardiography. AB - Murine models offer potential insights regarding human cardiac disease, but efficient and reliable methods for phenotype evaluation are necessary. We employed non-invasive electrocardiography (ECG) in mice, investigating statistical reliability of these parameters with respect to anesthetic and animal age. Mice (C57BL/6, 8 or 48 weeks) were anesthetized by ketamine/xylazine (K/X, 80/10 mg/kg ip) or by inhalation anesthetic (halothane, HAL; sevoflurane, SEV) and 6 lead ECGs were recorded. P wave duration and QT interval was significantly prolonged with K/X compared to HAL and SEV, indicating slowed atrial and ventricular conduction. P-R interval (atrio-ventricular conduction) was significantly increased in aged mice under all anesthetics. Heart rate was inversely correlated to QT interval and P wave duration. We also detected significant age effects with respect to optimal approaches for QT interval corrections. Power analysis showed 4-fold higher number of mice/group, were required for K/X, to achieve identical statistical sensitivity. These data demonstrate the importance of anesthetic selection for relevant and reliable ECG analysis in mice and illustrate the selective influences of anesthetics and age on cardiac conductance in this species. PMID- 12639706 TI - Regulation of hepatobiliary excretion of sinomenine by P-glycoprotein in Sprague Dawley rats. AB - Sinomenine, an herbal ingredient isolated from Sinomenium acutum, is used for the amelioration of arthritis. Using microdialysis and a specially constructed hepato duodenal shunt probe, the present study investigated the pharmacokinetics of sinomenine in rat blood and bile and the effects of P-glycoprotein modulation and cytochrome P450 inhibition. The results indicated that the pharmacokinetics of sinomenine in rat blood appeared to be dose dependent in the 3 to 30 mg/kg range. The disposition of sinomenine in the bile exhibited a slow elimination phase, reaching a peak concentration in 20-40 min following intravenous administration. The area under the concentration versus time curves (AUC's) for sinomenine in the bile were significantly greater than those in the blood at dosages of 3, 10, and 30 mg/kg with the blood-to-bile distribution ratios (k = AUC(bile) / AUC(blood)) being 3.85 +/- 0.29 and 3.52 +/- 0.28 at 10 and 30 mg/kg, respectively, indicating active hepatobiliary excretion. Coadministration with 20 mg/kg of cyclosporin A 10 min prior to sinomenine administration resulted in a significant reduction of the bile AUC's for the dosages of 10 and 30 mg/kg., resulting in the bile/blood distribution ratio being significantly reduced to 0.47 +/- 0.05 and 0.49 +/- 0.05, respectively. On the other hand, proadifen treatment increased both the blood and bile AUC's, resulting in insignificant effects on the blood-to bile distribution ratios. In conclusion, our results indicated that sinomenine underwent active hepatobiliary elimination which may be regulated by the P glycoprotein and that P-450 was likely involved in its metabolism. PMID- 12639709 TI - Mechanisms of regulating the Raf kinase family. AB - The MAP Kinase pathway is a key signalling mechanism that regulates many cellular functions such as cell growth, transformation and apoptosis. One of the essential components of this pathway is the serine/threonine kinase, Raf. Raf (MAPKK kinase, MAPKKK) relays the extracellular signal from the receptor/Ras complex to a cascade of cytosolic kinases by phosphorylating and activating MAPK/ERK kinase (MEK; MAPK kinase, MAPKK) that phosphorylates and activates extracellular signal regulated kinase (ERK; mitogen-activated protein kinase, MAPK), which phosphorylates various cytoplasmic and nuclear proteins. Regulation of both Ras and Raf is crucial in the proper maintenance of cell growth as oncogenic mutations in these genes lead to high transforming activity. Ras is mutated in 30% of all human cancers and B-Raf is mutated in 60% of malignant melanomas. The mechanisms that regulate the small GTPase Ras as well as the downstream kinases MEK and extracellular signal regulated kinase (ERK) are well understood. However, the regulation of Raf is complex and involves the integration of other signalling pathways as well as intramolecular interactions, phosphorylation, dephosphorylation and protein-protein interactions. From studies using mammalian isoforms of Raf, as well as C. elegans lin45-Raf, common patterns and unique differences of regulation have emerged. This review will summarize recent findings on the regulation of Raf kinase. PMID- 12639708 TI - Molecular recognitions in the MAP kinase cascades. AB - The mitogen-activated protein kinase (MAPK) cascades play a pivotal role in many aspects of cellular functions, and are evolutionarily conserved from yeast to mammals. In mammals, there are four subfamily members in the MAPKs. Each MAPK has its own activators, substrates and inactivators. In order to achieve normal cellular functions, the MAPK cascades should transduce signals with high efficiency and fidelity. However, the molecular basis for the mechanism underlying the specific reactions in the MAPK cascades has not been fully understood. The MAPKs form a globular structure without a distinct domain specific for protein-protein interactions. Recent studies revealed two mechanisms regulating the signalling, the docking interaction and the scaffolding. The docking interaction is achieved through the common docking domain (the CD domain) on MAPKs, and is different from a transient enzyme-substrate interaction through the active centre of the enzymes. Almost all the MAPK-interacting molecules have a conserved motif interacting with the CD domain. The scaffolding usually utilizes a third molecule to tether several components of the MAPK cascades. Both of them are thought to regulate the enzymatic specificity and efficiency. PMID- 12639711 TI - Interaction of Gbeta3s, a splice variant of the G-protein Gbeta3, with Ggamma- and Galpha-proteins. AB - The T-allele of a polymorphism (C825T) in the gene of the G-protein beta3-subunit is associated with a complex phenotype (hypertension, obesity, altered drug responses) and the occurrence of a splice variant termed Gbeta3s which lacks one of the seven WD-domains that compose Gbeta-proteins. Here, we analysed Gbetagamma dimer formation and Galpha activation by Gbeta3s, key functional characteristics of Gbeta-proteins. Cleavage protection assays frequently used to analyse Gbeta1gamma and Gbeta2gamma dimer formation failed for Gbeta3 and Gbeta3s, while in coprecipitation assays, dimerization of Gbeta3 and Gbeta3s with Ggamma5, Ggamma8(c) and Ggamma12 could be demonstrated. Upon expression of Gbeta3s in COS 7 and Sf9 insect cells, binding of GTPgammaS to Galpha-proteins induced by mastoparan-7 and the M(2) muscarinic acetylcholine receptor was facilitated in comparison with cells overexpressing wildtype Gbeta3, as indicated by twofold reduced agonist EC(50) values. Together, these results indicate that Gbeta3s is a biologically active Gbeta-protein that may mediate the enhanced signal transduction observed in cells with the 825T-allele. PMID- 12639710 TI - Calcineurin activates NF-kappaB in skeletal muscle C2C12 cells. AB - Calcineurin (CnA) is an important signalling molecule in skeletal muscle, in the promotion of differentiation, slow-fibre phenotype and possibly fibre hypertrophy. We found that stable expression of constitutively active CnA in muscle C2C12 cells strongly activated NF-kappaB, a key mediator of muscle wasting. NF-kappaB activation by CnA was associated with elevated phospho IkappaBalpha, and could be repressed by specific genetic (porZAKI-4 and porDSCR1) and chemical (cyclosporin A) inhibitors of CnA, but tumour necrosis factor-alpha (TNF-alpha) appeared not to be a key component in the cross-talk. Functionally, CnA-induced NF-kappaB activation seemed to interfere with terminal muscle differentiation. We therefore showed a functional interaction between the CnA and NF-kappaB pathways in skeletal muscle cells, which involved opposing phenotypic effects of CnA. PMID- 12639712 TI - Caveolar compartmentation of caspase-3 in cardiac endothelial cells. AB - Endothelial cell apoptosis is intimately involved in the balance between blood vessel growth and regression and is promoted by numerous stimuli including angiostatin and endostatin, reactive oxygen species (ROS) released during inflammatory processes, and chronic use of drugs of abuse such as cocaine. Apoptosis is characterized by many biological signalling events, including the activation of caspases. Caveolar domains have been hypothesized to mediate apoptotic signalling. We have addressed this hypothesis in cardiac endothelial cells and here we show that caspase-3 proenzyme (32 kDa) and its activated counterpart (17 kDa) co-purify with low-density, caveolin-enriched microdomains and that caspase-3 can be localized with caveolae in intact cells using fluorescent microscopy. Disruption of caveolae results in temporal and spatial changes in enzyme activity. While caspase-3 has been associated with mitochondrial, cytosolic, and high-density regions, the co-purification of activated caspase-3 and caveolar domains reported here suggests the possibility that sarcolemmal caspase-3 may be targeted to plasma-membrane associated substrates. PMID- 12639713 TI - Induction of cyclooxygenase-2 by lipopolysaccharide in canine tracheal smooth muscle cells: involvement of p42/p44 and p38 mitogen-activated protein kinases and nuclear factor-kappaB pathways. AB - Lipopolysaccharide (LPS) was found to induce inflammatory responses in the airways and exerted as a potent stimulus for PG synthesis. This study was to determine the mechanisms of LPS-enhanced cyclooxygenase (COX)-2 expression associated with PGE(2) synthesis in tracheal smooth muscle cells (TSMCs). LPS markedly increased the expression of COX-2 and release of PGE(2) in a time- and concentration-dependent manner, whereas COX-1 remained unaltered. Both the expression of COX-2 and the generation of PGE(2) in response to LPS were attenuated by a tyrosine kinase inhibitor genistein, a phosphatidylcholine phospholipase C inhibitor D609, a phosphatidylinositol-phospholipase C inhibitor U73122, protein kinase C inhibitors, GF109203X and staurosporine, removal of Ca(2+) by addition of BAPTA/AM plus EGTA, and phosphatidylinositol 3-kinase (PI3 K) inhibitors, LY294002 and wortmannin. Furthermore, LPS-induced NF-kappaB activation correlated with the degradation of IkappaB-alpha, COX-2 expression, and PGE(2) synthesis, was inhibited by transfection with dominant negative mutants of NIK and IKK-alpha, but not by IKK-beta. LPS-induced COX-2 expression and PGE(2) synthesis were completely inhibited by PD98059 (an inhibitor of MEK1/2) and SB203580 (an inhibitor of p38 MAPK inhibitor), but these two inhibitors had no effect on LPS-induced NF-kappaB activation, indicating that NF kappaB is activated by LPS independently of activation of p42/p44 MAPK and p38 MAPK pathways in TSMCs. Taken together, these findings suggest that the increased expression of COX-2 correlates with the release of PGE(2) from LPS-challenged TSMCs, at least in part, independently mediated through MAPKs and NF-kappaB signalling pathways. LPS-mediated responses were modulated by PLC, Ca(2+), PKC, tyrosine kinase, and PI3-K in these cells. PMID- 12639714 TI - Interleukin-8 production induced by the endozepine triakontatetraneuropeptide in human neutrophils: role of calcium and pharmacological investigation of signal transduction pathways. AB - The endozepine triakontatetraneuropeptide (TTN) induces intracellular calcium ([Ca(2+)](i)) changes and is chemotactic for human neutrophils (PMNs). Because interleukin-8 (IL-8) production is Ca(2+) dependent and can be induced by chemotactic stimuli, we have investigated the ability of TTN to induce IL-8 production in PMNs, as well as the signal transduction mechanisms involved. Our results show that TTN increases IL-8 release and IL-8 mRNA expression in a concentration- and time-dependent fashion, and these effects are prevented by the Ca(2+) chelator BAPTA-AM. TTN-induced [Ca(2+)](i) changes and IL-8 mRNA expression are sensitive to pertussis toxin, to the phospholipase C (PLC) inhibitor U73122 (but not to its inactive analogue U73343) and to the protein kinase C (PKC) inhibitor calphostin C. It is therefore suggested that TTN-induced IL-8 production in human PMNs results from a G protein-operated, PLC-activated [Ca(2+)](i) rise, and PKC contributes to this effect. These findings further support the possible role of TTN in the modulation of the inflammatory processes. PMID- 12639715 TI - Activation of B2 bradykinin receptors by neurotensin. AB - Many previous reports suggested that relatively high concentrations of neurotensin were required to exert its effects on neurotransmitter secretion. The neurotensin binding sites, which recognize high concentrations of neurotensin, were characterized in rat pheochromocytoma (PC12) cells. When PC12 cells were treated with neurotensin, [3H]norepinephrine secretion and elevation of cytosolic calcium were evoked at EC(50) values of 59+/-4 and 37+/-7 microM, respectively. Both calcium release and inositol 1,4,5-trisphosphate (IP(3)) production induced by neurotensin suggested involvement of phospholipase C. Experiments with simultaneous or sequential treatment with neurotensin and bradykinin suggested that neurotensin and bradykinin act on the same binding sites. Furthermore, both inhibition of bradykinin- and neurotensin-induced calcium rises by bradykinin receptor antagonists with similar IC(50) values and receptor binding analysis using [3H]bradykinin confirmed that neurotensin directly binds to B2 bradykinin receptors. The data suggest that neurotensin binds and activates the B2 bradykinin receptors. PMID- 12639716 TI - Protein kinase A- and C-induced insulin release from Ca2+ -insensitive pools. AB - Insulin secretion is known to depend on an increase in intracellular Ca(2+) concentration ([Ca(2+)](i)). However, recent studies have suggested that insulin secretion can also be evoked in a Ca(2+)-independent manner. In the present study we show that treatment of intact mouse islets and RINm5F cells with protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) or protein kinase A (PKA) activator forskolin promoted insulin secretion with no changes of [Ca(2+)](i). Moreover, insulin secretion mediated by PMA or forskolin was maintained even when extracellular or cytosolic Ca(2+) was deprived by treatment of cells with ethylene glycol bis(beta-amino ethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) or 1,2-bis(2-amino phenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis(acetoxy methyl ester) (BAPTA/AM) in RINm5F cells. The secretagogue actions of PMA and forskolin were blocked by GF109203X and H89, selective inhibitors for PKC and PKA, respectively. PMA treatment caused translocation of PKC-alpha and PKC- epsilon from cytosol to membrane, implying that selectively PKC-alpha and PKC- epsilon isoforms might be important for insulin secretion. Co treatment with high K(+) and PMA showed a comparable level of insulin secretion to that of PMA alone. In addition, PMA and forskolin evoked insulin secretion in cells where Ca(2+)-dependent insulin secretion was completed. Our data suggest that PKC and PKA can elicit insulin secretion not only in a Ca(2+)-sensitive manner but also in a Ca(2+)-independent manner from separate releasable pools. PMID- 12639717 TI - Survival pathways regulating the apoptosis induced by tumour necrosis factor alpha in primary cultured bovine endothelial cells. AB - The aim of the present study was to identify biochemical pathways driving the resistance of endothelial cells to apoptosis induced by tumour necrosis factor alpha (TNF). (1) Although nuclear factor-kappa B (NF-kappaB) was activated by TNF, its inhibition by MG-132 failed to sensitize these cells. (2) The activation of protein kinase C (PKC) by phorbol ester completely abolished the TNF-induced cell death. (3) The phosphatidylinositol 3-kinase (PI3K) inhibitor wortmannin (Wo) triggered apoptosis and enhanced the TNF-induced cell death. (4) The MEK inhibitor PD98059 did not affect the TNF-induced apoptotic process. (5) The p38 is activated by TNF and its inhibition by SB203580 sensitized the cells to TNF. This is correlated with the inhibition of phosphorylation of heat-shock protein of 27 kDa (HSP27). These results indicate that TNF activates NF-kappaB, which does not drive any anti-apoptotic response, and p38, which plays an anti apoptotic function probably through HSP27 phosphorylation. Moreover, PKC and PI3K are involved in the control of survival pathways. PMID- 12639718 TI - Serum and urine nitrate and nitrite are not reliable indicators of intrathecal nitric oxide production in acute brain injury. AB - This study examined the correlation between nitric oxide (NO) metabolites in the three major body fluid compartments and assessed performance of newly described vanadium-based assay for simultaneous detection of nitrite and nitrate (NO(x)) in human samples. Vanadium reduces nitrate to nitrite, which can be measured after a colorimetric reaction with Griess reagents. Cisternal cerebro spinal fluid (CSF), serum and urine samples from 10 patients with acute brain injury (ABI) were compared to control subjects. Significantly higher CSF NO(x) levels were found in brain injury patients compared to control patients (19.7+/-13.7 vs. 6.5+/-2.3 microM; p=0.01), which persisted for 10-day period of observation. The serum and urine levels of NO(x) on admission were not statistically different (42.8+/-28.2 microM; 584.1+/-337.8 micromol/g Cr, respectively) from controls (36.8+/-14.8 microM; 819.7+/-356.0 micromol/g Cr), but tended to decrease during the disease course reaching the lowest level on day 6 (serum: 19.3+/-8.4 microM, urine: 300.4+/-111.9 micromol/g Cr). CSF levels of NO(x) correlated moderately with those in serum (p=0.001, R=0.5). Serum NO(x) concentrations correlated weakly with urine levels (p=0.04, R=0.3). There was no significant correlation between CSF NO(x) and urine NO(x) levels. In conclusion, patients suffering brain injury had increased NO(x) concentrations in CSF, which remained independent from other body fluid compartments. Serum and urinary NO(x) levels cannot be used as a reliable index to assess intrathecal NO production. PMID- 12639719 TI - Protection of dopaminergic neurons with a novel astrocyte modulating agent (R)-( )-2-propyloctanoic acid (ONO-2506) in an MPTP-mouse model of Parkinson's disease. AB - We examined the neuroprotective effects of a novel astrocyte-modulating agent, (R)-(-)-2-propyloctanoic acid (ONO-2506), in a mouse model of Parkinson's disease. Male C57BL/6 mice received four intraperitoneal injections of 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP) (10 mg/kg) at 1-h intervals. Dopamine content in the striatum, measured with HPLC 3 days after MPTP injection, was reduced to 23% of control. But this dopamine depletion was dose-dependently prevented by repeated treatments with ONO-2506 (3, 10 and 30 mg/kg, i.p.) administered 1, 6, 24 and 48 h after MPTP injection (51% of control in 30 mg/kg group, p<0.01). ONO-2506 treatment (30 mg/kg) started after 6 h, followed by treatments at 24 and 48 h, also prevented the reduction of dopamine content (42% of control vs. 11% of control in the saline-treated group, p<0.01). We also performed immunohistochemistry for tyrosine hydroxylase (TH) and glial fibrillary acidic protein (GFAP). The MPTP injection resulted in a loss of TH-positive dopaminergic neurons (42% of control, p<0.01) in the substantia nigra after 7 days, but ONO-2506 treatment prevented this neuronal loss (70% of control, p<0.01). The MPTP injection led to reactive astrocytosis in the striatum after 7 days, but ONO-2506 induced earlier, moderate astrocytic activation after 3-7 days. These findings show that ONO-2506 protects dopaminergic neurons against MPTP neurotoxicity probably through facilitating astrocytic support for neuronal recovery from injury. Pharmacological modulation of astrocytes may offer a novel therapeutic strategy for Parkinson's disease. PMID- 12639720 TI - Association of phosphorylation site of tau protein with neuronal apoptosis in Alzheimer's disease. AB - In addition to neuritic changes and amyloid deposits, neuronal and glial cell apoptosis is an important pathological feature of Alzheimer's disease (AD). Several factors have been postulated as causes or triggers of cellular apoptotic change. This study focused on a quantifiable relationship between phosphorylation sites of tau protein in the neurofibrillary tangles (NFT) and neuronal apoptosis. Five monoclonal anti-tau antibodies (AT180, AT8, HT7, Tau2 and Tau5) for NFT labeling and TdT-mediated UTP nick-end labeling (TUNEL) for localizing apoptotic change were employed. TUNEL-stained neuronal nuclei showed significantly high density in the entorhinal cortex, cornu ammonis (CA) and the parietal cortex. In all regions, density of TUNEL-stained neuronal nuclei showed significantly direct correlation with that of AT8-, AT180- and Tau2-positive neurons. Correlation of TUNEL-stained neuronal nuclei with tau-positive neurons differed depending on the cerebral regions. Density of TUNEL-stained neuronal nuclei showed inverse correlation with that of both AT8-positive and Gallyas-stained NFT in the CA and showed significantly direct correlation with AT8- and HT7-positive neurons in the frontal cortex. Density of tau-positive and Gallyas-stained NFT was higher than that of TUNEL-stained nuclei. We conclude that phosphorylation sites of tau, 159 163 and 202-205, are probably associated with neuronal apoptosis and apoptotic change follows abnormal phosphorylation of tau. PMID- 12639721 TI - Predictors of gastrointestinal bleeding in acute intracerebral haemorrhage. AB - BACKGROUND: Gastrointestinal (GI) haemorrhage is an important and sometimes serious complication in critically ill neurological patients who suffered from stroke and head injury and those in intensive care. There is no study evaluating frequency, severity and risk factors of GI haemorrhage in patients with primary intracerebral haemorrhage (ICH). AIMS: To evaluate the frequency, severity and predictors of GI haemorrhage in patients with ICH. METHODS: In a prospective hospital-based study, consecutive CT-proven ICH patients within 10 days of the ictus were included. The patients with history of peptic ulcer, GI haemorrhage, liver and kidney disease, bleeding diathesis and those on antiplatelet, anticoagulant or nonsteroidal antiinflammatory drugs (NSAIDS) were excluded. A detailed neurological evaluation was carried out. Glasgow coma scale (GCS) was used for assessment of consciousness level and Canadian neurological scale (CNS) for severity of stroke. The haematomas were classified into small (<20 ml), medium (20-40 ml) and large (>40 ml). The occurrence of GI haemorrhage during 14 days of ictus was considered due to ICH. To evaluate the predictors of GI haemorrhage, various clinical and CT scan findings were evaluated by univariate followed by multivariate logistic regression analysis. RESULTS: Fifty-one patients with ICH were included whose age ranged between 30 and 80 years and 14 were female. The mean GCS score was 8.9 (3-15) and CNS score was 2.2 (2-4). Haematoma was small (<20 ml) in 11 patients and medium (20-40 ml) and large (>40 ml) in 20 patients each. Evidences of septicemia were present in 20 patients. Gastric haemorrhage (GH) was noted in 15 patients which was more than 40 ml in 4 patients and one of these patients needed blood transfusion. On univariate analysis, the size of haematoma, septicemia, motor signs on the nonhemiplegic side and pupillary asymmetry were significantly related to GI haemorrhage. On multivariate analysis, the best set of predictors of gastric haemorrhage included size of haematoma, septicemia and GCS score. CONCLUSION: GI haemorrhage is more likely present in patients with larger haematoma having septicemia. Our study highlights the importance of septicemia, which is an important and modifiable risk factor for GI bleeding in ICH patients. PMID- 12639722 TI - Calculation impairment in neurodegenerative diseases. AB - We examined oral calculation in patients with corticobasal degeneration (CBD; N=17), frontotemporal dementia (FTD; N=17), and Alzheimer's disease (AD; N=20), as well as 17 healthy seniors matched for age and education. Our calculation model involves at least three components: numerosity, combinatorial processes, and executive resources such as working memory. We assessed addition, subtraction, multiplication, and division involving small numbers (small, single digit answers) and large numbers (larger, single- and double-digit answers). We also assessed dot counting for small numbers (2-5) and large numbers (6-9), as well as a measure of working memory. All patient groups differed from healthy seniors in oral calculation. CBD (36% correct) and FTD (65% correct) demonstrated a significant overall impairment in oral calculation relative to AD (76% correct). CBD (66% correct) had more difficulty counting dots overall relative to AD (94% correct) and FTD (86% correct), consistent with our hypothesis that the calculation deficit in CBD is due in large part to a numerosity deficit. FTD had more difficulty relative to AD in their performance of reverse digit span, consistent with our hypothesis that FTD patients' executive resource limitation contributes to their pattern of calculation impairment. PMID- 12639723 TI - HIV-associated Guillain-Barre syndrome. AB - Human immunodeficiency virus-associated Guillain-Barre syndrome (HIV-GBS) has been reported since 1985. Based on previous reports, this neuropathy typically occurs early in HIV infection, even at seroconversion, prior to developing acquired immunodeficiency syndrome (AIDS). Patients with GBS and CD4 counts of <50 have been proposed to have cytomegalovirus (CMV) infection and empiric gancyclovir is recommended. We reviewed medical records of 10 patients with HIV GBS at five hospitals from 1986 to 1999. The mean CD4 count was 367/mm(3) (range 55-800). GBS was the first symptom of HIV infection in three patients. Four patients had AIDS with CD4 counts ranging from 55 to 190. CSF white blood cell (WBC) was 0 wbc/mm(3) in four patients, 2-10 wbc/mm(3) in three and 11-17 wbc/mm(3) in two. Three had recurrent weakness from 9 weeks to 4 years after the onset of symptoms, which persisted. HIV-GBS occurs in early and late stages of HIV infection, and may follow the onset of AIDS. No patients were seen with severe immunosuppression (CD4<50). A mild cerebrospinal fluid (CSF) pleocytosis in GBS suggests HIV infection, but is frequently absent. Compared to HIV-negative people, HIV-GBS may be associated with more frequent recurrent episodes or the development of CIDP. PMID- 12639724 TI - Changes in serum macrophage-related factors in patients with chronic inflammatory demyelinating polyneuropathy caused by intravenous immunoglobulin therapy. AB - Chronic inflammatory demyelinating polyneuropathy (CIDP) is a slowly progressive or recurrent neuropathy accompanied by infiltration of macrophages in the peripheral nerves. Macrophage colony-stimulating factor (M-CSF) and monocyte chemoattractant protein-1 (MCP-1) are a macrophage-related cytokine and chemokine, respectively. Although, intravenous immunoglobulin (IVIg) infusion therapy has been used for treating CIDP patients, not all CIDP patients have responded to IVIg infusion therapy. To determine the mechanisms of the action of IVIg, we examined serum M-CSF and MCP-1 levels during and after IVIg infusion therapy in 19 CIDP patients treated with IVIg (0.4 g/kg/day for 5 days). Ten of the 19 patients (52.6%) responded to IVIg therapy. Both M-CSF and MCP-1 concentrations in IVIg responders were significantly higher on day 1 postinfusion than those in nonresponders, but decreased to their pretreatment values on day 5 postinfusion. The results suggest that immunomodulation through M-CSF and MCP-1 is involved in the mechanisms underlying the effect of IVIg infusion therapy in CIDP patients. PMID- 12639726 TI - A semiautomated measure of whole-brain atrophy in multiple sclerosis. AB - Brain atrophy is a proposed MRI marker of irreversible pathologic damage in multiple sclerosis (MS). The brain parenchymal fraction (BPF) is the ratio of brain parenchymal volume to the total volume within the surface contour. We developed a semiautomated measure of BPF using commercially available edge finding and thresholding software (30-min analysis time per patient). We measured BPF in 78 patients with MS and 17 healthy controls. BPF was lower in a cohort of patients with MS (n=50) (0.843+/-0.042, range 0.743-0.906) age-matched to controls (0.877+/-0.020, range 0.835-0.901) (p<0.001). BPF correlated inversely with third ventricular width (r=-0.785, p<0.001), and total T1 hypointense lesion volume (r=-0.347, p=0.011), but not with total T2 hyperintense lesion volume (r= 0.213, p=0.13). BPF correlated negatively with expanded disability status scale (EDSS) score (r=-0.391, p=0.0006) and disease duration (r=-0.281, p=0.01). Stepwise regression compared the relative abilities of MRI variables to predict clinical data. By regression of age, BPF, third ventricular width, T2 lesions, and T1 lesions, BPF was the best predictor of disability score (R(2)=0.204, p<0.001). Third ventricular width was the best predictor of disease duration (R(2)=0.316, p<0.001). None of the MRI variables differed between relapsing remitting (RR) (n=60) and secondary progressive (SP) (n=18) disease course (p>0.05). The intrarater, interrater, and scan-rescan BPF variability (COV) was 0.31%, 0.34%, and 0.41% and the accuracy against a phantom was 99.1%. We conclude that whole-brain atrophy in MS can be reliably and readily quantified by a semiautomated approach. Longitudinal studies are warranted to determine if this method provides a sensitive biologic marker of the MS disease process. PMID- 12639725 TI - Dual impairment of GABAA- and GABAB-receptor-mediated synaptic responses by autoantibodies to glutamic acid decarboxylase. AB - Anti-glutamate decarboxylase autoantibodies (GAD-A) are associated with a group of patients with progressive cerebellar ataxia. We reported previously that cerebellar GABA(A)-mediated synaptic transmission was presynaptically depressed by GAD-A in the cerebrospinal fluid (CSF). Using whole-cell recording of rat cerebellar slices, we found in the present study that CSF immunoglobulins from ataxic patients reduced gamma-aminobutyric acid (GABA) release from cerebellar interneurons, thereby attenuating presynaptic inhibition on neighboring excitatory synapses through GABA(B) receptors (GABA(B)Rs). Our results suggest that in in vitro slices, GAD-A elicited the pathophysiological action of reduction in GABA release, which subsequently resulted in dual synaptic impairment in the cerebellar circuit, by depression of GABA(A) receptor (GABA(A)R)-mediated inhibitory synaptic transmissions, and attenuation of GABA(B) receptor-mediated inhibition of excitatory transmissions. PMID- 12639727 TI - Decreased galectin-1 immunoreactivity of the skin in amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease involving motor neurons. In addition to motor neuron signs and symptoms, a lack of bedsores has been considered a feature of ALS. Recently, we revealed that galectin-1 is a component of the axonal spheroid, which is an early pathological change of the spinal cord in ALS. To investigate whether galectin-1 is associated with skin changes in ALS, we performed an immunohistochemical investigation using anti-galectin-1 antibodies. The present study revealed that galectin-1 immunoreactivity is reduced in the skin of patients with ALS, suggesting that cutaneous galectin-1 is involved in the pathological process of ALS. PMID- 12639728 TI - Assessment of cerebrovascular and cardiovascular responses to lower body negative pressure as a test of cerebral autoregulation. AB - The aim of this study was to determine whether lower body negative pressure (LBNP), combined with noninvasive methods of assessing changes in systemic and cerebral vascular resistance, is suitable as a method for assessing cerebral autoregulation. In 13 subjects we continuously assessed heart rate, blood pressure, cerebral blood flow velocity (CBFV) and cardiac output during graded levels of LBNP from 0 to -50 mm Hg. With increasing levels of LBNP, cardiac output declined significantly (to 55.8+/-4.5% of baseline value) but there was no overall change in mean arterial pressure. CBFV also fell at higher levels of LBNP (to 81.4+/-3.2% of baseline) but the percentage CBFV change was significantly less than that in cardiac output (P<0.01). The maximum increase in cerebrovascular resistance (pulsatility ratio) was significantly less than that in total peripheral resistance (17+/-6% vs. 105+/-16%, P<0.01). Spectral analysis showed that the power of low-frequency oscillations in mean arterial pressure, but not CBFV, increased significantly at the -50 mm Hg level of LBNP. These results show that, even during high levels of orthostatic stress, cerebral autoregulation is preserved and continues to protect the cerebral circulation from changes in the systemic circulation. Furthermore, assessment of cardiovascular and cerebrovascular parameters during LBNP may provide a useful clinical test of cerebral autoregulation. PMID- 12639729 TI - Prolonged peripheral nerve stimulation induces persistent changes in excitability of human motor cortex. AB - This study sought to determine whether prolonged peripheral nerve stimulation was effective in inducing persistent "plastic" changes in the excitability of the human motor cortex. The amplitude of the electromyographic response evoked in resting intrinsic hand muscles by focal transcranial magnetic stimulation (TMS) was taken as an index of motor cortical excitability. Twelve subjects were stimulated with each of three protocols, one of which was given on each of three separate occasions. The protocols consisted of various schedules of electrical stimulation of the radial and ulnar nerves or the motor point of the first dorsal interosseous muscle (FDI), or stimulation of FDI motor point paired with low frequency TMS. Amplitudes of TMS-elicited motor evoked potentials (MEPs) were measured before peripheral stimulation and for 2 h after stimulation. The data from one subject were unusable. In every other subject, all three protocols induced a prolonged, significant facilitation of MEPs in at least some of the three intrinsic hand muscles used. In some instances, MEPs were not enlarged and occasionally were significantly depressed. Different protocols based on peripheral afferent stimulation can induce plastic changes in the organisation of the motor cortex that persist for at least 2 h. PMID- 12639730 TI - Unusual clinical findings and Complex III deficiency in a family with myotonic dystrophy. AB - Myotonic dystrophy type 1 (DM1), an autosomal dominant disease characterized by a CTG expansion in the 3' region of the DMPK gene in chromosome 19, is a highly heterogeneous disease. In this study, we present a family with early onset classical type DM, and a homogeneous phenotype highlighted by severe neuromuscular symptoms and mental dysfunction with subcortical-type dementia. Neuroradiological abnormalities included brain atrophy, white matter lesions, and basal ganglia calcifications. A very large CTG triplet expansion was present in the DMPK locus in leukocytes in the three patients. One patient showed ragged-red fibers (RRF) and a defect complex III of the respiratory chain, but no mutations were found in the cytochrome b gene of mtDNA. PMID- 12639731 TI - Hypertensive brainstem encephalopathy: clinical and radiographic features. AB - Reversible posterior leukoencephalopathy syndrome associated with hypertension rarely presents with predominant involvement of the brainstem and relative sparing of the supratentorial regions. We describe the clinical and neuroimaging features of three patients with reversible hypertensive brainstem encephalopathy. Headache and confusion in the setting of severe hypertension, with a relative paucity of brainstem signs and symptoms, despite extensive neuroimaging abnormalities in the brainstem support the diagnosis. The clinical findings and neuroimaging abnormalities resolve with treatment of the hypertension, further distinguishing this syndrome from brainstem infarction. PMID- 12639732 TI - Subacute brainstem angioencephalopathy: a case report and review of the literature. AB - A previously healthy 69-year-old man developed a progressive neurological illness with bulbar signs and ataxic paraparesis. Repeated MRI examinations revealed a large space occupying lesion in the lower brain stem with patchy contrast enhancement. MRI angiography was unremarkable and CSF had normal cell count but raised protein content. A brainstem tumor was suspected and a course of intravenous glucocorticosteroids was started. No improvement occurred and the patient died of pneumonia 11 weeks after onset. Neuropathology revealed confluent areas of complete or incomplete necrosis with marked edema in the lower brainstem. Predominantly venous meningeal vessels of the brainstem showed extensive fibromuscular thickening of all layers with luminal narrowing. In addition, intramural mononuclear infiltration was found. With the exception of localisation, this case exhibits all pathologic features of subacute diencephalic angioencephalopathy (SDAE), a rare fatal disease of unknown aetiology. In addition, the clinical features of typical age, male sex, disease duration and raised CSF proteins are shared. A common disease entity is suggested and the pathogenetic relevance of inflammation and venous outflow obstruction is discussed. PMID- 12639733 TI - Statin therapy and small fibre neuropathy: a serial electrophysiological study. AB - We describe three patients who developed small fibre neuropathy after 1 month of statin therapy with clinical resolution upon prompt drug withdrawal. All patients showed abnormal sympathetic skin responses (SSR) in comparison with controls. SSRs returned to normal in tandem with clinical improvement. One patient redeveloped small and large fibre neuropathy when the similar drug was readministered. The SSR is of value in the electrophysiological assessment and follow-up of statin-related small fibre neuropathy. PMID- 12639734 TI - Hemichorea: a rare presentation of tuberculoma. AB - Hemichorea due to tuberculoma has not been reported. We managed a 16-year-old girl who developed transient left hemichorea due to right sided striatal tuberculoma. Chorea regressed following 1 week of antitubercular treatment with corticosteroid therapy and disappeared at 3 months. In a patient presenting with chorea, striatal tuberculoma should also be considered especially in the area where tuberculosis is prevalent. PMID- 12639735 TI - Sarcoidosis of the cauda equina mimicking Guillain-Barre syndrome. AB - Neurosarcoidosis is a great mimicker. It is often difficult to diagnose particularly when there is no prior history of systemic sarcoidosis. Although certain sites of the neuraxis are more commonly involved than others, any site of the central or peripheral nervous system can be affected. We report a case of sarcoidosis involving the cauda equina in a 38-year-old African American male without prior history of systemic disease. Initial clinical presentation was suggestive of Guillian-Barre syndrome, but the evaluation proved this case to be neurosarcoidosis involving the cauda equina. We have followed this patient for 8 years, and he remains clinically stable on prednisone 5 mg/day. PMID- 12639738 TI - Cultural categorization of febrile illnesses in correlation with herbal remedies used for treatment in Southwestern Nigeria. AB - The ethnographic study was conducted in two communities in Oyo State in Southwestern Nigeria. The study sites consisted of a rural and an urban local government area located in the tropical rain forest zone of Nigeria. The study was designed to obtain information on febrile illnesses and herbal remedies for treatment with the aim of identifying potential antimalarial drugs. The study revealed that fever is a general term for describing illnesses associated with elevated body temperature. The indigenous Yoruba ethnic population has categorized fever based on symptoms and causes. The present communication is the result of focus group discussion and semi-structured questionnaire administered to traditional healers, herb sellers, elders and mothers. This was on types of fevers, symptoms and causes of febrile illnesses. The investigation also included use of traditional herbs in the prevention and treatment of the illnesses in the two communities.A total of 514 respondents were interviewed. This was made up of 266 (51.8%) from Atiba local government area (LGA), an urban centre while 248 (48.2%) respondents were interviewed from Itesiwaju LGA, a rural community. The LGAs are located in Oyo State of Nigeria. The respondents proffered 12 types of febrile illnesses in a multiple response answering system in Yoruba language. The most common ones (direct translation into English) were: yellow fever (39.1%), typhoid (34.8%), ordinary (28.8%), rainy season (20.8%) and headache (10.5%) fevers, respectively. Perceived causes of each of the febrile illnesses included stress, mosquito bites, unclean water, rains and over exposure to the sun. Methods of fever prevention were mainly with the use of herbal decoctions, powdered herbs, orthodox medications and maintenance of proper hygiene. Of a total of 112 different herbal remedies used in the treatment of the febrile illnesses compiled from the study, 25 recipes are presented. Recipes consisted of 2-7 ingredients. Oral decoctions (84%), oral powders (63%), use as soaps and creams (40%) in a multiple response system, were the most prevalent routes of administration of prepared herbs used in the treatment of the fevers. Boiling in water or alcohol was the most common method used in the preparation of the remedies. The four most frequently mentioned (multiple response system) plants in the Southwest ethnobotany for fevers were Azadirachta indica (87.5%), Mangifera indica (75.0%), Morinda lucida (68.8%) and Citrus medica (68.8%). PMID- 12639739 TI - Anti-inflammatory activity of 'TAF' an active fraction from the plant Barleria prionitis Linn. AB - 'TAF' fraction from the methanol-water extract of Barleria prionitis Linn. was evaluated for anti-inflammatory and anti-arthritic activities against different acute and chronic animal test models. It exhibited significant anti-inflammatory activity against different inflammagens like carrageenan, histamine and dextran. The anti-inflammatory activity in adrenalectomised rats was maintained showing that the effect of fraction 'TAF' is not activated by the pituitary-adrenal axis. Significant anti-arthritic activity was observed in adjuvant-induced polyarthritis test in rats. 'TAF' also showed inhibition of vascular permeability and leucocytes migration in vivo into the site of inflammatory insult.Ibuprofen (Cadilla India Ltd., Mumbai) was used as a standard reference drug. The oral (p.o.) LD(50) was more than 3000mg/kg, with no signs of abnormalities or any mortality observed for 15 days after single-dose drug administration. The intraperitoneal (i.p.) LD(50) was found to be 2530mg/kg (+/-87mg/kg S.E.) [Proceedings of Society of Experimental Biological Medicine 57 (1944) 261]. PMID- 12639740 TI - Effects of Fagonia cretica L. constituents on various haematological parameters in rabbits. AB - Effects of powdered Fagonia cretica plant and its two major triterpenoid saponins (saponin-I and saponin-II), isolated from its ethanolic extract, on red blood cells (RBC) count, haemoglobin concentration (HC), mean corpuscular haemoglobin (MCH) and on total leucocyte (WBC) count of normal male rabbits were investigated. Saponins treated three dose groups of animals indicated significant decrease in RBC count during the experimental period of 16 days. This effect was more pronounced in animals treated with saponin-II than saponin-I. The 0.50 and 1.0g crude drug treated animals and 10 and 20mg of both the saponins treated animals, indicated a decreasing tendency of HC up to the 4th day, which increased afterwards up to the 16th day. The third dose of these materials (1.50g of crude drug and 30mg of both saponins) exhibited a highly significant decreasing pattern, up the 16 days. The decreasing effect of haemoglobin concentration was more distinct in the saponin-II treated animals than saponin-I and the crude drug. The MCH followed a reverse pattern than RBC and HC. A continued decreasing trend was found in total WBC during the 16 days treatment. An amount of 1.50g of the crude drug and 30mg of both the saponins had highly significant decreasing effects on the amount of total leukocyte count of rabbit's blood. PMID- 12639741 TI - Hypoglycaemic and hypolipidemic effect of ethanolic extract of seeds of Eugenia jambolana in alloxan-induced diabetic rabbits. AB - The hypoglycaemic and hypolipidemic effect of ethanolic extract obtained from seeds of E. jambolana was investigated in alloxan-induced diabetic rabbits. Hypoglycaemic activity was assessed by reduction in fasting blood glucose (FBG) at 90min and also fall in peak blood glucose during glucose tolerance test (GTT) in sub-diabetic and mild diabetic (MD) rabbits, but in severe diabetic (SD) rabbits by reduction in FBG at 90min. Ethanolic extract (100mg/kg body weight) when given orally to sub-diabetic (AR) for 1 day, MD for 7 days and SD for 15 days showed significant fall in FBG at 90min (12% AR, 18.9% MD and 29% SD) and also produced 16.9% fall in peak blood glucose in AR and 21% in MD rabbits during GTT. When administered daily for 15 days to MD and SD rabbits, significant fall in FBG (41.3% MD, 31.6% SD) and glycosylated haemoglobin (GHb) levels (23.3% MD, 26.6% SD) were observed, while serum insulin level showed significant increase (32.8% MD, 26.9% SD). Liver and muscle glycogen content also increased. The ethanolic extract of seeds also exhibited significant hypolipidemic effect as evident from fall in total serum cholesterol (TC)/high density lipoprotein cholesterol (HDL-c) ratio, serum low density lipoprotein cholesterol (LDL-c) levels and decreased activity of HMG-CoA reductase. The histopathological studies of liver, pancreas and aorta in alcoholic extract treated diabetic groups revealed almost normal appearance. PMID- 12639742 TI - An evaluation of the activity related to inflammation of four plants used in Thailand to treat arthritis. AB - The leaves of Acanthus ebracteatus, stembark of Oroxylum indicum and the stems of Cryptolepis buchanani and Derris scandens are used as traditional remedies in Thailand for arthritis. Aqueous and alcoholic extracts were tested using three different in vitro systems for effects relevant to anti-inflammatory activity. The aqueous extracts of O. indicum and D. scandens significantly reduced myeloperoxide release. Eicosanoid production was reduced only by the aqueous extracts of A. ebracteatus and D. scandens. D. scandens extract showed potent inhibitory activity against generation of leukotriene B(4) and also displayed antioxidant activity. In the rat hind paw edema test, D. scandens extract showed significant activity when given intraperitoneally but did not produce a significant reduction when given orally. The results therefore supported to some extent the traditional use of D. scandens for arthritic conditions and provided slight indication of activity which could explain the use of O. indicum and A. ebracteatus. No relevant activity was demonstrated in any of the tests for C. buchanani extracts. PMID- 12639743 TI - Evaluation of the anti-inflammatory properties of Sclerocarya birrea (A. Rich.) Hochst. (family: Anacardiaceae) stem-bark extracts in rats. AB - This study was designed to evaluate the anti-inflammatory effect of Sclerocarya birrea (family: Anacardiaceae) stem-bark aqueous and methanolic extracts in rats. Young adult, male Wistar rats (Rattus norvegicus) weighing 250-300g were used. The anti-inflammatory effects of aqueous and methanolic stem-bark extracts of the plant (SB, 500mg/kg p.o.) were examined on rat paw oedema induced by subplantar injections of fresh egg albumin (0.5ml/kg). Acetylsalicylic acid (ASA, 100mg/kg p.o.) was used as the reference anti-inflammatory agent for comparison. Both the aqueous and methanolic stem-bark extracts of S. birrea (SB, 500mg/kg p.o.) progressively and time-dependently reduced rat paw oedema induced by subplantar injections of fresh egg albumin. However, the methanolic extract of the plant produced relatively greater and more pronounced anti-inflammatory effect than its aqueous extract counterpart in the experimental animal model used. The two extracts of S. birrea stem-bark were found to be markedly less potent than ASA as anti-inflammatory agent. Although both the aqueous and methanolic extracts of S. birrea stem-bark are less potent than ASA as anti-inflammatory agent, the results of this experimental animal study indicate that the extracts possess anti inflammatory activity, and thus lend credence to the suggested folkloric use of the plant in the management and/or control of arthritis and other inflammatory conditions in certain communities of South Africa. PMID- 12639744 TI - The analgesic, antipyretic and anti-inflammatory activity of Diospyros variegata Kruz. AB - Pharmacological studies were conducted with the hexane extract of the dry stem of Diospyros variegata Kruz. (Ebenaceae) on experimental animals for evaluating the analgesic, antipyretic and anti-inflammatory activities. In the analgesic test, the hexane extract elicited inhibitory intensity on acetic acid-induced writhing response and on the late phase of formalin test but possessed only a weak effect on the tail-flick response and on the early phase of formalin test. The hexane extract also elicited antipyretic action when tested in yeast-induced hyperthermia in rats. In addition, the hexane extract showed an anti-inflammatory effect when tested in ethyl phenylpropiolate (EPP)- and arachidonic acid (AA) induced rat ear edema. PMID- 12639745 TI - Antioxidant activity of the ethanolic extract of Striga orobanchioides. AB - Plants containing flavonoids have been reported to possess strong antioxidant properties. The ethanolic extract of Striga orobanchioides was screened for in vitro and in vivo antioxidant properties using standard procedures. The ethanol extract exhibited IC(50) values of 18.65+/-1.46 and 11.20+/-0.52 micro g/ml, respectively in DPPH and nitric oxide radical inhibition assays. These values were less than those obtained for ascorbic acid and rutin, used as standards. In the in vivo experiments the extract treatment at 100mg/kg body weight dose caused a significant increase in the level of the catalase in the liver and the kidneys. A significant increase in the level of SOD in the liver was observed. The treatment also caused a significant decrease in the TBA-RS and increase in the ascorbic acid levels. These results suggest strong antioxidant potentials of the ethanolic extract of S. orobanchioides. PMID- 12639746 TI - Antimicrobial activity of aqueous and methanol extracts of Juniperus oxycedrus L. AB - Aqueous and methanol extracts of the leaves of Juniperus oxycedrus were investigated for their in vitro antimicrobial properties. The plant was collected from Pelitli Village of Gebze, Kocaeli, in the Marmara region of Turkey. Juniperus oxycedrus is widely used as traditional folk medicine in Turkey for treatment of different infectious diseases. The antimicrobial activity of the extracts against 143 laboratory strains belonging to 56 bacterial species, and 31 isolates of 5 fungi species were evaluated based on the inhibition zone using the disc-diffusion assay, minimal inhibition concentration (MIC) and minimal bactericidal concentration (MBC) values. The aqueous extract of J. oxycedrus had no antimicrobial effect against the test microorganisms whereas the methanol extract had inhibitory effects on the growth of 57 strains of 24 bacterial species in the genera of Acinetobacter, Bacillus, Brevundimonas, Brucella, Enterobacter, Escherichia, Micrococcus, Pseudomonas, Staphylococcus, and Xanthomonas. In addition 11 Candida albicans isolates at a concentration of 31.25 250 micro g/ml were also inhibited. PMID- 12639748 TI - Ethnomedical field study in northern Peruvian Andes with particular reference to divination practices. AB - The results of a field study carried out in August and September 1988, in the northern Peruvian Andes are described. The area of investigation extends from Ayabaca City (about 2900m above sea level) to Haughtiness Lagunas (about 3800m above sea level) in the Ayabaca District, Department of Piura. This is the first time that this location has been the subject of an ethnobotanical investigation. We have collected 46 plant species, belonging to 20 families, used in the treatment of various diseases. For each plant, we report the common/local names, the crude drug formulation, method of preparation, dosage and claimed toxicity. The disease concept of this Andean population concerning the "hot" and "cold" aspects of diseases and the plants to treat them, is also discussed. Very important appear to be the use and knowledge of psychoactive plants, in particular "cimoras," Brugmansia and Trichocereus species. PMID- 12639747 TI - Effect of T. foenumgraecum on glycogen content of tissues and the key enzymes of carbohydrate metabolism. AB - The Indian traditional system of medicine prescribed plant therapies for diseases including diabetes mellitus called madhumeh in Sanskrit. One such plant mentioned in Ayurveda is Trigonella foenumgraecum (FG). In the present study, FG (1g/kg PO) was assessed for its effect on glycogen levels of insulin dependent (skeletal muscle and liver), insulin independent tissues (kidneys and brain) and enzymes such as glucokinase (GK), hexokinase (HK), and phosphofructokinase (PFK). Administration of FG led to decrease in blood glucose levels by 14.4 and 46.64% on 15th and 30th day of the experiment. Liver and 2-kidney weight expressed as percentage of body weight was significantly increased in diabetics (P<0.0005) versus normal controls and this alteration in the renal weight (P<0.0005) but not liver weight was normalized by feeding of FG. Renal glycogen content increased by over 10 folds while hepatic and skeletal muscle glycogen content decreased by 75 and 68% in diabetic controls versus controls and these alteration in glycogen content was partly prevented by FG. Activity of HK, GK and PFK in diabetic controls was 35, 50 and 60% of the controls and FG partially corrected this alteration in PFK, HK and GK. PMID- 12639749 TI - Tribulus terrestris: preliminary study of its diuretic and contractile effects and comparison with Zea mays. AB - OBJECTIVES: Tribulus terrestris L. (Zygophyllaceae) which is called Al-Gutub (in Iraqi dialect) or Quti;ba (in classical Arabic medicine), and Zea mays were both used alone or in combination by Iraqi herbalists to propel urinary stones. We studied the aqueous extract of the leaves and fruits of T. terrestris and the hair of Z. mays, to determine their diuretic activity and the contractile effect of T. terrestris. METHODS: The aqueous extract was filtered and the solvent was evaporated to produce a dry crude extract. The dry extract was then dissolved in physiological saline to make the required concentrations. Wistar male rats were used for the diuresis test and strips of isolated Guinea pig ileum were used for the contractility test. RESULTS: The aqueous extract of T. terrestris, in oral dose of 5g/kg elicited a positive diuresis, which was slightly more than that of furosemide. Z. mays aqueous extract did not result in significant diuresis when given alone in oral dose of 5g/kg, while combination of Z. mays and T. terrestris extracts produced the same extent of diuresis as that produced by T. terrestris alone. Na(+), K(+) and Cl(+) concentrations in the urine had also much increased. In addition to its diuretic activity T. terrestris had evoked a contractile activity on Guinea pig ileum. CONCLUSION: T. terrestris has long been used empirically to propel urinary stones. The diuretic and contractile effects of T. terrestris indicate that it has the potential of propelling urinary stones and merits further pharmacological studies. PMID- 12639750 TI - Anti-inflammatory and analgesic activity in the polyherbal formulation Maharasnadhi Quathar. AB - Maharasnadhi Quathar (MRQ) is a polyherbal preparation recommended by Ayurvedic medical practitioners for treatment of arthritic conditions. An investigation has been carried out with rats and human rheumatoid arthritis (RA) patients, to determine the anti-inflammatory and analgesic potential of MRQ. Results obtained demonstrate that MRQ can significantly and dose-dependently inhibit carrageenan induced rat paw oedema (the inhibition at 3h was greater than at 1h after induction of oedema). MRQ could also increase the reaction time of rats in the hot-plate test (by 57% after the first hour of treatment), although it had no effect on the reaction time in the tail-flick test, indicating that MRQ possesses analgesic activity that is probably mediated via a supra-spinal effect.MRQ also exerted a dose-dependent (a) protective effect on heat-induced erythrocyte lysis, and (b) inhibition of 5-lipoxygenase activity. In RA patients, after 3 months of MRQ treatment, there was a marked improvement in the pain and inflammation experienced by the patients as well as in the mobility of the affected joints. From the overall results obtained, it may be concluded that MRQ possesses significant anti-inflammatory and analgesic activities. Alteration in synthesis of prostaglandins and leukotrienes, membrane stabilization and anti-oxidant activity are some of the possible mechanisms through which MRQ mediates its anti arthritic effects. PMID- 12639751 TI - Medicinal use and social status of the soap berry endod (Phytolacca dodecandra) in Ethiopia. AB - Berries from Phytolacca dodecandra L'Herit. (endod in Amharic) offer a readily available molluscicide to control schistosomiasis. Parts of the endod plant have been used as a detergent and as traditional medicine for centuries in Ethiopia. An interview survey was performed in the highlands of Ethiopia to provide information on the distribution of the plant, people's traditional use of it, their perception of the plant, and the potential for increased production and use of endod as a soap for indirect control of schistosomiasis. People of all ages report that they are familiar with the plant and its detergent and medicinal uses. The plant is largely disappearing from unprotected areas due to land clearing. Younger people appear to use endod as a soap whenever it is available. Older women prefer commercial soap and consider endod to be associated with poor people. Common medicinal uses include treatment of skin itching (ringworm), abortion, gonorrhea, leeches, intestinal worms, anthrax and rabies. Two thirds of the people express interest in cultivating endod for personal use if supplied with rooted cuttings. Increased cultivation of endod and use of berries for washing might be possible if information about schistosomiasis and its control is disseminated among people. Preference for commercial soap and lack of land for cultivation are major obstacles for increasing the availability and use of endod. PMID- 12639752 TI - Tigliane diterpenes from the latex of Euphorbia obtusifolia with inhibitory activity on the mammalian mitochondrial respiratory chain. AB - Six diterpenes isolated from the latex of Euphorbia obtusifolia Poir. (Euphorbiaceae) were evaluated for their inhibition of the NADH oxidase activity in submitochondrial particles from beef heart. 4,20-Dideoxyphorbol-12,13 bis(isobutyrate) was the most potent inhibitor and showed an inhibitory concentration with IC(50) value of 2.6+/-0.3mM. In the present study, some structure-activity trends are suggested for the inhibitory activity of the mammalian mitochondrial respiratory chain of these natural product derivatives of 4-deoxyphorbol esters. PMID- 12639753 TI - Inhibitory effect of aqueous extract from the gall of Rhus chinensis on alpha glucosidase activity and postprandial blood glucose. AB - The present study examined the inhibitory effect of aqueous extract from the gall of Rhus chinensis (AEGRC) on alpha-glucosidase activity, an enzyme responsible for digestion of carbohydrate to monosaccharides in the process of intestinal absorption. AEGRC inhibited Bacillus alpha-glucosidase acitvity with an IC(50) of 0.9 micro g/ml. Its inhibition on alpha-glucosidase was determined to be noncompetitive and reversible when the enzyme-substrate mixture was simultaneously treated with AEGRC as an inhibitor. In addition, when it was orally administered to rats with sucrose (2g/kg), AEGRC (250-1000mg/kg) significantly suppressed the increase of blood glucose levels after sucrose loading in a dose dependent manner. These results suggest that AEGRC might exert anti-diabetic effect by suppressing carbohydrate absorption from intestine, and thereby reducing the postprandial increase of blood glucose. PMID- 12639754 TI - Inhibition of human pathogenic fungi by members of Zingiberaceae used by the Kenyah (Indonesian Borneo). AB - Extracts from 11 plant species belonging to the Zingiberaceae were tested for antifungal activity using disc diffusion bioassays. Extracts from several members, especially Alpinia galanga, Curcuma zedoaria and Zingiber purpureum, were found to have pronounced inhibitory activities against a wide variety of human pathogenic fungi, including strains resistant to the common antifungals amphotericin B and ketoconazole. As members of the Zingiberaceae are generally regarded as safe for human consumption, these species are excellent candidates for development as novel therapeutics. PMID- 12639759 TI - The role of polyol pathway in glucose-induced apoptosis of cultured retinal pericytes. AB - The pathogenesis of pericyte loss, an initial deficit in the early stage of diabetic retinopathy, remains unclear. Recent studies have suggested that polyol pathway hyperactivity and apoptosis may be involved in pericyte loss. The mechanisms of the glucose-induced apoptosis in retinal pericytes were investigated to evaluate the pathogenesis of diabetic retinopathy. Under the 20 mM glucose condition, intracellular calcium concentrations and caspase-3 activities were significantly increased, and reduced glutathione (GSH) contents were significantly decreased compared with those under the 5.5 mM glucose condition. These abnormalities were all significantly prevented by an aldose reductase inhibitor, SNK-860. Glucose-induced apoptosis was partially but significantly prevented by SNK-860, an inhibitor of calcium-dependent cysteine protease, calpain, or GSH supplementation, and completely normalized by a caspase 3 inhibitor. These observations suggest that glucose-induced apoptosis in retinal pericytes, as one of the pathogenic factors of diabetic retinopathy, would be mediated through an aldose reductase-sensitive pathway including calcium-calpain cascade and increased oxidative stress, and that caspase-3 would be located furthest downstream of these apoptotic signals. PMID- 12639760 TI - Enhanced vascular production of superoxide in OLETF rat after the onset of hyperglycemia. AB - This study was aimed to characterize the vascular production of superoxide in Otsuka Long-Evans Tokushima Fatty (OLETF) rat, a model of type 2 diabetes. The nitroblue tetrazolium staining in the aorta from old (30 weeks) OLETF rat was more prominent than that of age-matched control (LETO) rat, which was significantly inhibited by diphenyleneiodonium (10 micromol/l), but not by inhibitors for other oxidases such as xanthine oxidase, mitochondrial oxidase, nitric oxide synthase, and cyclooxygenase. In the aorta from old OLETF rat with hyperglycemia, the enhanced NADH oxidase activity in association with upregulated expression of p22phox and gp91phox was observed, but not in both LETO and young (10 weeks) OLETF rats without hyperglycemia. Furthermore, there was a positive correlation (P<0.01) between elevation of blood glucose level and increase in vascular NADH oxidase activity. Based on these results, it was suggested that the enhanced NADH oxidase activity in the aorta from OLETF rat occurred after the onset of hyperglycemia, thereby resulting in the increased vascular production of superoxide. PMID- 12639761 TI - Insulinoma accompanied by diabetes mellitus. AB - A 53-year-old type 2 diabetic man was admitted due to spontaneous relatively hyperinsulinemic hypoglycemia. Oral glucose ingestion and arginine tolerance test showed hyperinsulinemic response. Arterial stimulation and venous sampling (ASVS) showed hyperinsulinemic response measured from the splenic artery after calcium gluconate stimulation. Diagnosis was insulinoma in the pancreas feeding from the artery. He has not suffered from spontaneous hypoglycemia since removal of the pancreatic body, tail and spleen. The specimen showed a solitary islet cell tumor. The high homeostasis model assessment of insulin resistance (HOMA-R) levels reflecting insulin resistance and hyperinsulinemic response after operation remained almost unchanged, indicating high insulin resistance and an insulin hypersecreting diabetic patient. PMID- 12639762 TI - A case showing an association between type 1 diabetes mellitus and Kabuki syndrome. AB - The case of a 31-year-old female suffering from type 1 diabetes mellitus (DM) and Kabuki syndrome is presented. The patient was diagnosed as having impaired glucose tolerance (IGT) at age 18; secondary amenorrhea occurred at age 20, following acute body weight loss. Extensive examination revealed the patient to have a slowly progressive form of type 1 DM and, based on the physical findings, including her facial features, she was diagnosed as also having congenital Kabuki syndrome. Since then, this patient has experienced several episodes of diabetic ketoacidosis, all of which were brought about by prolonged bronchial infection. Although it is perhaps reasonable at present to consider this case to represent a chance association, further clinical investigations will be carried out to clarify whether or not Kabuki syndrome and type 1 DM have any common pathogenic features. PMID- 12639763 TI - HRT does not improve urinary albumin excretion in postmenopausal diabetic women. AB - The effect of 6 months combined, continuous hormone replacement therapy (HRT) with conjugated equine oestrogen (0.625 mg) and medroxyprogesterone acetate (2.5 mg) on albumin/creatinine ratio (ACR) was determined in postmenopausal diabetic women in a randomised, controlled study. Mean (interquartile range) change in plasma ACR was not (P=0.96) different in women receiving HRT [2 (-11, 21) mg/g, n=20] compared with those randomised to placebo [2 (-1, 14) mg/g, n=27]. Also, the proportion of women with microalbuminuria did not change (P=0.75) during HRT (baseline, 0.45; end of study, 0.53). Furthermore, several risk factors for microalbuminuria including systolic blood pressure (SBP), fasting blood glucose, glycated haemoglobin (HbA1c) and adiposity did not vary significantly during HRT. These data suggest that 6 months HRT does not reverse microalbuminuria caused by prolonged hyperglycaemia and other risk factors that underlie leakage of albumin into the urine in postmenopausal women with type 2 diabetes. PMID- 12639764 TI - Factors affecting perinatal morbidity and mortality in pregnancies complicated by diabetes mellitus in Sudan. AB - To investigate the influence of obstetric factors and indices of maternal metabolic control on perinatal morbidity and mortality, 88 diabetic pregnant Sudanese women (type 1, n=38; type 2, n=31; gestational diabetes, n=19) and 50 non-diabetic pregnant control women were studied. The mean fasting blood glucose was 11.1+/-2.8 mmol/l and the mean HbA(1c) at booking interview was 8.8+/-2.1% in the diabetic women. Pregnancy complications such as Caesarean sections, urinary tract infections, pregnancy-induced hypertension and intrauterine foetal death were higher among diabetic compared with control women (P<0.0001) and varied with the type of diabetes. Infants of diabetic mothers had a higher incidence of neonatal complications than those of non-diabetic women (54.4% vs. 20.0%; P<0.0001). Infants without complications and who were born to diabetic mothers had better Apgar scores at 5 min (9.8+/-0.5 vs. 8.9+/-1.6; P<0.01) and lower cord C-peptide when compared to infants with complications (P<0.05). In conclusion, the prevalence of maternal and neonatal complications among Sudanese diabetic women and their infants is high. Maternal hyperglycaemia is an important factor affecting maternal wellbeing and neonatal morbidity and mortality. PMID- 12639765 TI - NEUROD polymorphism Ala45Thr is associated with Type 1 diabetes mellitus in Czech children. AB - Association of the NEUROD Ala45Thr polymorphism with Type 1 diabetes mellitus (DM) has been found in some but not all populations. We performed a study on the association of two NEUROD exon 2 polymorphisms, the Ala45Thr and the Pro197His, with childhood-onset Type 1 DM in the Czech population. We compared 285 children with Type 1 DM diagnosed under the age of 15 years with 289 non-diabetic control children. The genotypes were determined using novel real-time allele-specific PCR assays in the TaqMan format, and data were analysed using logistic regression. The numbers of subjects with codon 45 genotypes Ala/Ala, Ala/Thr, Thr/Thr were 95, 145, 45 among cases and 117, 130, 42 among controls. Thr45 phenotypic positivity was associated with a significant risk of Type 1 DM (OR=2.01, CI 95% 1.25-3.24) in a multivariate logistic regression model involving also the insulin gene -23HphI genotype and the presence of Type 1 DM-associated HLA-DQB1*0302 DQA1*03 (DQ8) and DQB1*0201-DQA1*05 (DQ2) molecules. No association was observed for the Pro197His mutation which was carried by 5.3% cases and 5.9% controls. Our results confirm that the NEUROD Ala45Thr polymorphism is associated with childhood-onset Type 1 DM. PMID- 12639766 TI - Association of limited joint mobility and high plantar pressure in diabetic foot ulceration in Asian Indians. AB - AIM: To investigate the association of limited joint mobility (LJM) and plantar foot pressure in Asian Indian diabetic subjects. SUBJECTS AND METHODS: The LJM and plantar pressure were measured in 345 consecutive subjects attending the foot clinic. The study groups were: Control-non-diabetic controls (n=50), DM-diabetic patients without neuropathy (n=100), DM+N-diabetic neuropaths (n=110) and DM+NU diabetic neuropaths with past history of foot ulceration (n=85). Joint mobility was assessed using a goniometer. Plantar pressure was measured using the RS-Scan platform system. All subjects were able to walk comfortably unaided at their own pace. Data obtained on the metatarsal heads were used for analysis. Neuropathy status was assessed using the biothesiometer. RESULTS: The diabetic patients had higher prevalence of LJM and higher plantar pressure than control subjects (P<0.0001). Among the diabetic patients, those with neuropathy and history of plantar ulceration had higher LJM and plantar pressure compared to non-neuropaths (P<0.001). CONCLUSIONS: Both LJM and high plantar pressure appear to be important determinants of foot ulceration in susceptible neuropathic South Indian diabetic patients. PMID- 12639767 TI - Cutoff value of 1 h, 50 g glucose challenge test for screening of gestational diabetes mellitus in a Japanese population. AB - A total of 2651 consecutive native Japanese women who underwent a glucose challenge test (GCT) were retrospectively investigated. GCT was performed between 24 and 27 weeks of gestation; each subject received a 50 g oral glucose load without regard to the fasting or fed state, followed by a determination of 1 h venous plasma glucose level. Women demonstrating GCT exceeding 130 mg/dl received a 75 g, 2 h oral glucose tolerance test to determine whether or not they had gestational diabetes mellitus (GDM). All women with GDM were treated with a strict diabetic protocol including insulin therapy. Forty-nine (1.8%) women were diagnosed to have GDM. The receiver-operator characteristic curve identified a GCT finding above 140 mg/dl as the cutoff value for detecting GDM, which showed a sensitivity and specificity of 96 and 76%, respectively. Our results suggest that the cutoff value of a 50 g GCT is 140 mg/dl to identify pregnancies with GDM in a Japanese population. PMID- 12639768 TI - Type-1 diabetes mellitus with insufficient serum immunoreactive insulin elevation after subcutaneous NPH-insulin injection. AB - We report a case of Type-1 diabetes with insufficient serum immunoreactive insulin (IRI) elevation after subcutaneous NPH-insulin injection. Favorable glycemic control was achieved by a continuous subcutaneous insulin infusion (CSII) using regular insulin. A 34-year-old woman with Type-1 diabetes (height 158 cm, weight 43.4 kg) was admitted to our hospital to improve glycemic control. On admission, her glycosylated hemoglobin (HbA(1c)) level was 10.9% and her fasting plasma glucose (FPG) level was 332 mg/dl. After admission, her insulin regimen was altered from two injections a day using premixed insulin to four injections a day using regular insulin before each meal and NPH insulin at bedtime. Although the dosage of NPH insulin at bedtime was increased to 32 U/day, there was no improvement in the FPG level. The peak IRI value after NPH insulin injection was not observed but that after the regular insulin injection was observed. Therefore, her insulin administration regimen was changed to CSII, using regular insulin alone. Her fasting plasma glucose level decreased, glycosylated hemoglobin (HbA(1c)) level improved to 7.0%, her body weight increased to 46.6 kg 4 months after starting CSII. PMID- 12639769 TI - Is high diabetes mortality in Taiwan due to coding artifact? Implications for international comparisons. PMID- 12639770 TI - Hippocampal malrotation with normal corpus callosum in a child with Opitz syndrome. AB - We present cranial magnetic resonance imaging (MRI) findings in a 6-year-old boy with Opitz syndrome. The electroencephalogram revealed epileptiform abnormalities in both frontal regions. MRI showed normal callosal development, but a rounded shape of the left hippocampus due to malrotation. The temporal horn was slightly dilated on the left side. These features were consistent with isolated hippocampal malrotation. PMID- 12639771 TI - Magnetic resonance imaging of the intratemporal facial nerve in idiopathic peripheral facial palsy. AB - The aim of this study was to investigate the prevalence of facial nerve involvement with gadolinium-enhanced magnetic resonance imaging (Gd-MRI) in patients with idiopathic peripheral facial palsy (IPFP), and to discuss the localization and the pattern of enhancement. A total of 13 patients (9 female, 4 male) with IFPF were included in this study. Topographic tests and electromyography (EMG) were performed, and MRI was taken. Ten subjects whose cranial MRIs were taken for nonorganic pathology served as the control group. Twelve of 13 paralytic facial nerves had enhancement on postcontrast images. Two facial nerves of the control group demonstrated enhancement. We found a correlation between the enhancement of the facial nerve and the time for recovery. The average time from the onset of facial palsy to the recovery in patients with enhancement was 14 weeks, whereas it was 6 weeks in patient with no enhancement. Finally, all patients had complete recovery of the facial nerve function. We concluded that contrast enhancement of the paralytic facial nerve can be a radiological sign of a neural inflammation and may indicate a prolonged recovery. PMID- 12639772 TI - Juxtapapillary diverticulum: findings on CT and MRI. AB - The purpose of this study was to evaluate the computed tomography (CT) and magnetic resonance imaging (MRI) features of juxtapapillary diverticulum. CT and/or MRI examinations of 12 patients were evaluated. The size, location of the diverticula and imaging findings of associated pancreaticobiliary disease were assessed. On CT, oral contrast air level was the characteristic imaging feature. On MRI, T2-weighted images demonstrated the air fluid level with hyperintense fluid and signal void air level above. Associated imaging findings were dilated common bile duct (CBD), cholecystitis, cholecystolithiasis and chronic pancreatitis. PMID- 12639773 TI - Tumor and tumor-like lesions of duodenum: CT and barium imaging features. AB - Neoplasms of the duodenum, either primary or secondary, are uncommon. However, imaging diagnosis for presurgical evaluation is extremely important due to complex regional anatomy. Computed tomography (CT) and barium-based double contrast examination (UGI series) are most frequently employed to evaluate the tumor invasion and intraluminal mucosal pattern. In this pictorial review, the CT and UGI series imaging features of benign and malignant duodenal tumors and tumor like lesions are demonstrated and discussed. PMID- 12639774 TI - Hepatopulmonary syndrome: angiography and therapeutic embolization. AB - We report a case where an embolization therapy was successfully performed for diffusely dilated pulmonary vessels in a patient with hepatopulmonary syndrome (HPS) as a palliative treatment before liver transplantation. To our knowledge, this therapeutic approach has been tried only for arteriovenous malformation type HPS, and not for diffuse dilatation [Chest 91 (1987) 920]. PMID- 12639775 TI - Portal venous system: evaluation with contrast-enhanced 3D MR portography. AB - The purpose of this study is to compare contrast-enhanced three-dimensional (3D) magnetic resonance (MR) portograms to Doppler sonography in detection of portal venous abnormalities. Thirty-five consecutive patients, who were suspected of having portal venous system abnormalities, were examined with MR portography and Doppler sonography. Vascular abnormalities were identified in 27 of 35 patients. There was statistically significant agreement between the results of MR portography and Doppler sonography. The major limitation of contrast-enhanced 3D MR portography was its inability to provide objective hemodynamic data regarding flow direction and flow pattern. PMID- 12639776 TI - Solid, cystic and vanishing tumors of the pancreas. AB - Two girls with pancreatic tumors are reported. They were presented with abdominal pain, no history of abdominal injury and no tumor marker abnormalities. Imaging studies demonstrated that the tumors in two patients had both solid and cystic components. The tumors gradually regressed and finally disappeared without any treatment. There has been no evidence of recurrence in 5- and 4-year observation periods, respectively. Pancreatic tumor with solid and cystic components may rarely be self-limited. PMID- 12639777 TI - Fat necrosis mimicking liposarcoma in a patient with pelvic lipomatosis. CT findings. AB - Pelvic lipomatosis is a rare condition characterized by an overgrowth of normal fat in the perirectal and perivesical spaces. The most important differential diagnosis is liposarcoma. We present a case of pelvic lipomatosis associated with a mass, which was diagnosed radiologically as a liposarcoma, but surgical biopsy result revealed fat necrosis. PMID- 12639778 TI - Comparison of reliabilities of the delayed images of helical renal CT in detecting small renal masses. AB - The reliabilities of the delayed images of helical computed tomography (CT) in detecting renal mass lesions of 30 mm or less in diameter was compared. Nephrographic, excretory and nephrographic+excretory phase images of all patients were evaluated separately to detect mass lesions of < or = 5 and 5-30 mm. There was not any statistically significant difference in the results of three groups. Nephrographic, excretory and nephrographic + excretory phase images are not different from or superior to each other in lesion detection. PMID- 12639779 TI - CT findings of omental torsion and infarction: case report and review of the literature. AB - A 37-year-old woman presented with progressive diffuse abdominal pain. Computed tomography (CT) showed diffuse streaking of the greater omentum with a mass of fat density located anteriorly just below the umbilicus, showing a whirling pattern of concentric streaks. Surgery and pathology revealed torsion and infarction of the greater omentum. Because of its rarity and nonspecific clinical features, the diagnosis is seldom made preoperatively. We describe a patient with characteristic CT findings of omental torsion. Preoperative diagnosis is important since conservative management has been suggested. PMID- 12639780 TI - Multiplanar reformat display technique in abdominal multidetector row CT imaging. AB - The purpose of this study was to evaluate the image quality and diagnostic value of multiplanar reformat (MPR) of abdominal CT performed with multidetector row spiral technique (MDCT). Forty-five patients were referred for an abdominal MDCT angiography. No significant differences in image quality and in the number of lesions detected were found between axial and MPR images (t-test, P>.90 and chi square, P>.95). Additional diagnostic value to axial images could be obtained with MPR display. PMID- 12639781 TI - T2-weighted echo planar MR imaging of the abdomen: optimization of imaging parameters. AB - Echo planar imaging (EPI) is an ultra-fast magnetic resonance (MR) imaging method that provides strong T2-weighted contrast, which is important for the detection and characterization of various abdominal abnormalities. However, EPI may be associated with significant artifacts. The prominence of these artifacts, as well as overall image quality, is dramatically impacted by selection of imaging parameters. The purpose of this study is to demonstrate the effects of controlled alterations in operator-defined imaging parameters on image quality in breath hold abdominal T2-weighted EPI. An understanding of these effects facilitates optimizing parameter selection for acquisition of abdominal EPI images. PMID- 12639782 TI - Clinically occult isolated right iliac mycotic aneurysm with duodenal involvement in a diabetic elderly man: multislice CT diagnosis. AB - A diabetic old man presented with vague abdominal discomfort and intermittent tarry stools for 2 days and gastric ulcers with bleeding was diagnosed after endoscopy. Multislice computed tomography (MSCT) clearly depicted an isolated right iliac mycotic aneurysm with retroperitoneal extension and duodenal involvement. Timely operation and effective antibiotic treatment resulted in complete recovery. To our knowledge, this is the first report of an isolated mycotic iliac artery aneurysm (IAA) complicated with an aneurysmo-duodenal fistula induced by Klebsiella pneumoniae. PMID- 12639783 TI - MR-angiographic diagnosis of an aberrant retroaortic left renal vein and review of the literature. AB - MR-angiography is suitable for depiction of developmental vascular anomalies such as an aberrant left renal vein. We report MRA findings of one case of a retroaortic left renal vein and the review of the literature. PMID- 12639805 TI - Kinin B2 receptor localization and expression in the hypothalamo-pituitary adrenal axis of spontaneously hypertensive rats. AB - OBJECTIVE: An enhanced hypothalamo-pituitary-adrenocortical (HPA) activity has been demonstrated during onset of high blood pressure in spontaneously hypertensive rats (SHR). Furthermore, compared to normotensive Wistar-Kyoto (WKY) rats, SHR show hypersensitivity to bradykinin (BK)-induced pressor responses which may be caused by an upregulation of B(2) receptor expression in the brain. METHODS: We performed an immunohistochemical localization and measured gene expression of B(2) receptors in the hypothalamus, pituitary and adrenal glands of SHR at three ages corresponding to the development of hypertension, i.e. prehypertensive phase, onset of hypertension and established hypertension. Using reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot technique, B(2) receptor mRNA and protein levels, respectively, were measured. RESULTS: A specific immunostaining for B(2) receptors was observed in the hypothalamic nuclei paraventricularis (PVN) and supraopticus (SON). In the pituitary and adrenal glands, a strong immunostaining was observed in neurohypophysis (NH) and adrenal medulla, respectively. At all ages tested, B(2) receptor mRNA and protein levels were higher in the hypothalamus and adrenal glands of SHR compared to age-matched WKY rats. Among SHR, the mRNA level was increased in neurohypophysis with age, and no difference was found in the adenohypophysis (AH) between SHR and WKY rats. CONCLUSION: The data demonstrate a specific localization and an upregulation of B(2) receptor expression in the hypothalamus and adrenal glands of SHR, providing an anatomical and molecular basis for a possible contributory role to bradykinin-induced hypersensitivity of cardiovascular responses. The increased B(2) receptor expression in the hypothalamus and adrenal glands may also play a role in the abnormalities of the HPA axis in SHR during the development of hypertension. PMID- 12639806 TI - Effect of bradykinin and PMA on the synthesis of proteoglycans during the cell cycle of endothelial cells in culture. AB - Bradykinin (BK) and phorbol 12-myristate-13-acetate (PMA) were used in the present work to study the biosynthesis of proteoglycans (PG) during the cell cycle of endothelial cells. PMA, an activator of PKC, stimulated the synthesis of heparan sulfate proteoglycan (HSPG) secreted to the medium of endothelial cells mainly during the G(1) phase of the cell cycle [J. Cell. Biochem. 70 (1998) 563]. BK is a vasoactive peptide that increases calcium levels inside the cells indirectly stimulating PKC. Treatment of the endothelial cells with BK, as well as PMA, stimulated the synthesis of HSPG secreted to the medium and produced an antimitogenic effect on the cell cycle. These results led to the conclusion that PKC is directly involved in the synthesis of HSPG secreted to the medium. Also, comparing the effect showed by BK with PMA, one may suggest that different PKC isoforms are involved in these two processes and that their isoforms are mainly Ca(2+) dependent. PMID- 12639808 TI - Bradykinin and the pathophysiology of angioedema. AB - Angioedema has different causes and different clinical presentations. Some types of angioedema may be mediated by bradykinin. We measured plasma levels of bradykinin-(1-9)nonapeptide by radioimmunoassay after high-performance liquid chromatography in patients with different types of angioedema during acute attacks and/or in remission, i.e. hereditary C1-inhibitor deficiency, angiotensin converting enzyme (ACE) inhibitor treatment, idiopathic non histaminergic and responders to antihistamines. Eleven patients with the deficiency of C1-inhibitor had very high levels of bradykinin during acute attacks of angioedema (18.0-90.0 pM) (normal range 0.2-7.1 pM). In three patients with history of ACE inhibitor related angioedema, plasma bradykinin was high during ACE inhibitor treatment (62.0, 8.9 and 27.0 pM) and in a fourth patient was 47.0 pM during an acute attack and decreased by 93% to 3.2 pM after withdrawal of the ACE inhibitor. The patient with idiopathic angioedema, during an acute attack involving the right arm, had high levels of bradykinin in the venous blood refluent from the angioedematous arm (20.0 pM) while in the contralateral arm bradykinin levels were normal (6.6 pM), similarly to what we previously observed in cases of brachial angioedema due to C1-inhibitor deficiency. The four patients with angioedema responsive to antihistamines had normal levels of bradykinin even during acute attacks (5.7, 3.4, 4.7 and 1.2 pM). In one of these patients who had a brachial angioedema, bradykinin levels were normal in the venous blood refluent from both arms. Bradykinin is involved in hereditary C1-inhibitor deficiency angioedema, in ACE inhibitor-related angioedema, and in idiopathic non histaminergic angioedema, while bradykinin is not related to allergen-dependent or idiopathic angioedema that are responsive to antihistamines. PMID- 12639807 TI - Complement component C1q induces endothelial cell adhesion and spreading through a docking/signaling partnership of C1q receptors and integrins. AB - The interaction of C1q with endothelial cells elicits a multiplicity of biologic responses. Although these specific responses are thought to be mediated by the interaction of C1q with proteins of the endothelial cell surface, the molecular identity of the participant(s) has not been clearly defined. In this study, we examined the role of two C1q-binding proteins, cC1q-R/CR and gC1q-R/p33, on C1q mediated adhesion and spreading of human dermal microvascular endothelial cells (HDMVECs). A specific and dose-dependent adhesion and spreading was observed when HDMVECs were cultured in microtiter plate wells coated with concentrations of C1q ranging from 0 to 50 microg/ml. The extent of adhesion and spreading was similar to the adhesion seen on collagen-coated wells. Furthermore, the effect of C1q was mimicked by either polyclonal anti-cC1q-R or mAb 60.11, but not with isotype- and species-matched control IgG. More importantly, however, a 100% inhibition of spreading but not adhesion to C1q-coated wells was observed when HDMVECs were cultured in the presence of 30 mM of the peptide GRRGDSP but not GRRGESP. Furthermore, while anti-beta1 integrin antibody blocked adhesion and spreading, antialpha5 integrin only blocked spreading. Since earlier studies have shown that zinc induces the exposure of hydrophobic sites in the C-terminus of gC1q-R including the putative high-molecular weight kininogen (HK)-binding site corresponding to residues 204-218, we also examined the effect of zinc on antibody binding to cell surface gC1q-R. Flow cytometric data show that the binding of mAb 74.5.2, which recognizes residues 204-218, is greatly enhanced when endothelial cells were incubated in the presence of 50 microM zinc. In summary, our data show that: (a) C1q-mediated endothelial cell adhesion and spreading requires the cooperation of both C1q receptors and 1 integrins, and possibly other membrane-spanning molecules, and (b) zinc can induce the exposure of hydrophobic sites in the C-terminal domain of gC1q-R allowing a more efficient binding of mAb 74.5.2 and HK. PMID- 12639809 TI - Vascular permeability enhancement in solid tumor: various factors, mechanisms involved and its implications. AB - Most solid tumors are known to exhibit highly enhanced vascular permeability, similar to or more than the inflammatory tissues. Common denominators affecting both cancer and inflammatory lesions are now well known: bradykinin (BK), nitric oxide (NO), peroxynitrite (ONOO(-)), prostaglandins (PGs), collagenases or matrix metalloproteinases (MMPs) and others. Incidentally, enzymes involved in these mediator syntheses are upregulated or activated. Initially described vascular permeability factor (VPF) (proteinaceous) was later identified to be the same as vascular endothelial growth factor (VEGF), which promotes angiogenesis of cancer tissues as well. These mediators cross-talk or co-upregulate each other, such as BK-NO-PGs system. Therefore, vascular permeability observed in solid tumor may reflect the other side of the coin (angiogenesis). The vascular permeability and accumulation of plasma components in the interstitium described here is applicable for predominantly macromolecules (molecular weight, Mw>45 kDa), but not for low molecular compounds as most anticancer agents are. Macromolecular compounds (e.g., albumin, transferrin) or many biocompatible water-soluble polymers show this effect. Furthermore, they are not cleared rapidly from the sites of lesion (cancer/inflammatory tissue), thus, remain for prolonged time, usually for more than a few days. This phenomenon of "enhanced permeability and retention effect" observed in cancer tissue for macromolecules and lipids is coined "EPR effect", which is now widely accepted as a gold standard for anticancer drug designing to seek more cancer-selective targeting using macromolecular drugs. Consequently, drastic reduction of the systemic side effect is observed, while the macromolecular drugs will continuously exert antitumor activity. Other advantages of macromolecular drugs are also discussed. PMID- 12639810 TI - Evaluation of tissue kallikrein activity on survival time after acute coronary artery ligation in hypertensive rats. AB - It is known that the tissue kallikrein-kinin system is located in the cardiac tissue, and the lack of this system in the cardiac tissue might induce cardiac dysfunctions. In this study, we investigated the potential role of tissue kallikrein and Trasylol, an inhibitor of tissue kallikrein, on survival time with acute left coronary artery ligation for 15 min in spontaneously hypertensive rats (SHR). Tissue kallikrein (8 and 16 microg/kg, i.v.) treatment caused significant (P<0.05) increases in the survival time of SHR as compared with the saline treated control SHR. Trasylol pretreatment abolished (P<0.05) the beneficial effect on tissue kallikrein on survival time. The ligation of coronary artery resulted in significant (P<0.05) reduction in systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR) of SHR compared with the saline-treated control SHR. The tissue kallikrein treatment caused greater (P<0.001) reduction in the SBP, DBP and HR of SHR, when the mean values were compared between before coronary artery ligation and after coronary artery ligation. Trasylol (6 microg/kg) treatment antagonized the effects of tissue kallikrein associated with survival time, SBP, DBP and HR. These findings may suggest that tissue kallikrein is able to act as a cardioprotective agent as demonstrated by the increase in survival time of SHR with acute coronary artery ligation. The significance of these observations is discussed. PMID- 12639811 TI - Kinins are involved in the antiproteinuric effect of angiotensin-converting enzyme inhibition in experimental diabetic nephropathy. AB - The present study examined non-insulin-treated streptozotocin (STZ)-induced diabetic rats to determine the role of kinins in diabetic nephropathy. Their involvement in the renoprotective effect of the angiotensin-converting enzyme inhibitor (ACEI) ramipril was investigated using the bradykinin (BK) B(2) receptor antagonist, icatibant (HOE 140), or a combination of the two drugs.Although, none of the treatments prevented the decline of the glomerular filtration rate (GFR) in diabetic rats, ramipril (3 mg/kg/day), but not icatibant (HOE 140; 500 microg/kg/day), prevented proteinuria in these animals. However, the antiproteinuric effect of ramipril was reduced by 45% when combined with icatibant. To explore whether the renal kallikrein-kinin system (KKS) belongs to the underlying mechanisms of these findings, we also determined urinary BK levels, renal kallikrein (KLK) and angiotensin-converting enzyme (ACE) activity as well as renal cortical mRNA levels of neutral endopeptidase 24.11 (NEP) and low-molecular weight (LMW) kininogen. STZ led to a reduction of renal KLK and ACE activity and NEP expression and to a three-fold increase of urinary BK excretion and renal kininogen expression. Icatibant given alone had no effect on these parameters. In contrast, ramipril treatment normalized urinary protein and BK excretion as well as kininogen mRNA expression without affecting NEP mRNA expression or KLK and ACE activity. Our data demonstrate that renal BK is increased in severe STZ-induced diabetes mellitus, but may affect glomerular regulation only to a minor degree under this condition. However, kinins are partly involved in the antiproteinuric action of ACEI at this stage of diabetic nephropathy. PMID- 12639812 TI - Maturation-dependent expression of C1q-binding proteins on the cell surface of human monocyte-derived dendritic cells. AB - The expression and cell surface levels of many important receptors are dependent on the maturation stage of dendritic cells (DCs), and related to the unique function of immature and mature DCs. In this report, we show for the first time that human monocyte-derived DCs express two types of C1q-receptors, gC1qR and cC1qR. Furthermore, immature DCs secrete detectable amount of C1q into the culture supernatant. Immature DCs express higher cell surface levels of both C1qRs than mature ones, while the total C1qR protein and mRNA levels remain the same. The following experimental evidence supports this conclusion. (1) Inflammatory cytokines and LPS, which induce maturation of DCs, downregulate surface expression of both C1qR molecules. (2) Cytokines and drugs (IL-10, IFNalpha, dexamethasone) that keep DCs phenotypically and functionally immature significantly upregulate the cell surface expression of both C1qRs. (3) Neither of these treatments changed the intracellular gC1qR level nor the gC1qR mRNA levels measured by real-time RT-PCR. The elevated surface expression of C1qRs on DCs has been found not to be due to increased apoptosis or cell death as the result of DC treatment. Taken together, these data show that human monocyte derived DCs express gC1qR and cC1qR, their expression on the cell surface is maturation dependent and imature DCs secrete C1q. These data strongly suggest the role of C1qRs in immature DC function and in the regulation of immune processes. PMID- 12639813 TI - Reduced apoptosis by pentoxiphylline among patients with HIV infection. AB - BACKGROUND: Apoptosis is a significant cause of CD4(+) T cell death. Caspase 8 (FLICE) is involved in apoptosis mediated by Fas and p55 tumor necrosis factor (TNF) receptor ligation. It is also partially mediated by interleukin-1beta (IL 1beta)-converting enzyme (ICE; caspase 1). We and others have shown that pentoxiphylline inhibits TNF-alpha. We used it among patients with HIV infection to determine if 24 weeks of therapy altered the levels of caspase 1 and caspase 8. PATIENTS AND METHODS: Nineteen HIV-infected patients having no opportunistic infection at the time of recruitment were administered pentoxiphylline 400 mg thrice daily for 24 weeks. Caspase levels were measured using a single-step ELISA using commercially available monoclonal antibodies against caspase 1 and caspase 8. RESULTS: Mean CD4 counts of the patients were 202.6+/-111.6 (/mm(3)). Mean OD value of caspase 1 among patients before therapy was 0.302+/-0.197 and was higher than that of controls (0.287+/-0.064), but this was not statistically significant. Following 24 weeks of therapy with pentoxiphylline, the OD value declined significantly to 0.164+/-0.028 among patients (p<0.001). Mean OD value of caspase 8 among patients prior to therapy was 0.927+/-0.249. This was significantly higher than that of controls, whose level was 0.0074+/-0.004 (p<0.001). Following 24 weeks of therapy with pentoxiphylline, the OD value declined to 0.199+/-0.064 among patients and this was significantly lower than the value at the start of treatment (p<0.001). CONCLUSION: Therapy with pentoxiphylline for 24 weeks is associated with a decline in the levels of caspase 1 and caspase 8. Since the drug is known to produce TNF inhibition, this might result in reduced apoptosis and an improved CD4 lymphocyte survival. PMID- 12639814 TI - Suppressive effect of a traditional Japanese medicine, Hachimi-jio-gan (Ba-Wei-Di Huang-Wan), on the hyperresponsiveness to IL-18 in autoimmune MRL/MPJ-lpr/lpr mice. AB - Oral administration of Hachimi-jio-gan (HMG, Ba-Wei-Di-Huang-Wan), a traditional Japanese herbal medicine, for several weeks, ameliorates some autoimmune symptoms of MRL/lpr mice. In the short time treatment for 9 days, hyperresponsiveness of mesenteric lymph node (MLN) cells to interleukin (IL)-18 manifested by the proliferation or the production of interferon (IFN)-gamma was significantly suppressed. Additionally, the treatment with HMG suppressed the expressions of IL 18Ralpha and IL-18Rbeta mRNA in CD45R(-) T cells, and also the expression of IL 18Ralpha mRNA in unpurified whole cells. Although the short treatment with HMG had no effect on the mRNA expressions of IL-10, IL-12 and IL-18, or the phosphorelated signal transducer and activator transcription (STAT)4 protein level in CD45R(-) MLN cells, the IL-4 mRNA expression or the phosphorelated STAT6 protein level were up-regulated by HMG, and the IL-4 mRNA up-regulation was clearer in whole cells than CD45R(-) cells. Furthermore, the treatment with HMG promoted the mRNA expression of invariant Valpha14 TCR that is uniquely expressed on NKT cells. Valpha14 NKT cells can produce large amount of IL-4 and play a crucial role in controlling the development of MRL/lpr mouse autoimmune disease. Therefore, these results suggested that HMG reduced the hyperresponsiveness of MRL/lpr mouse MLN cells to IL-18 through the reduction of IL-18Rs caused by Valpha14 NKT cell-produced IL-4, and consequently HMG suppressed the development of MRL/lpr autoimmune diseases. PMID- 12639815 TI - Stimulation of murine macrophages by native and heat-denatured lectin from Abrus precatorius. AB - We have evaluated the immunostimulant activity of native agglutinin (NA) and heat denatured agglutinin (HDA) obtained from Abrus precatorius seed kernels on murine macrophages. Activated macrophages play an important role in nonspecific immunostimulation in turn by activating the other immune cells in the cascade. Significant alterations are observed in the macrophage functions both by native and heat-denatured Abrus agglutinin. Increased production of nitric oxide and hydrogen peroxide and a high phagocytic and bactericidal activity is potentiated by both NA and HDA. It is also observed that activated macrophages also released interleukin-1. These results suggest that both NA and HDA act as immunostimulants in vitro. PMID- 12639816 TI - In vitro activity of labdane diterpene from Alomia myriadenia (Asteraceae): immunosuppression via induction of apoptosis in monocytes. AB - A screening program in Brazilian flora was carried out to detect the presence of immunosuppressive compounds by using the in vitro phytohemagglutinin A (PHA) induced human peripheral blood mononuclear cell (PBMC) proliferation assay. In this screening, we isolated from Alomia myriadenia Schultz-Bip. ex. Baker (Asteraceae), a labdane-type diterpene named myriadenolide. Incubation of human PBMC with this compound reduced significantly the percentage of CD14(+) cells, but it has no effect on the relative amount of CD3(+)CD4(-)CD8(+) and CD3(+)CD4(+)CD8(-) T lymphocyte subpopulations. Neither viability nor proliferative competence of T lymphocytes was significantly affected by myriadenolide. The toxic effect on monocytes (CD14(+) cells) may explain the inhibitory effect observed on PHA-induced lymphocyte proliferation. The cytotoxic effect of myriadenolide on monocytes was determined by measuring the percentage of hypodiploid nuclei content by propidium iodide staining, electron microscopy and simultaneous detection of CD14 and annexin V binding by flow cytometry. The results showed that myriadenolide induces a dose-dependent apoptosis in monocytes and thus explain the immunosuppressive effect observed. PMID- 12639817 TI - Effect of dietary whey protein concentrate on primary and secondary antibody responses in immunized BALB/c mice. AB - Proteins derived from the whey fraction of bovine milk are known to modulate immune responses. We have previously described a rennet whey protein concentrate (WPC) that can boost intestinal tract antibody responses to orally administered T dependent antigens. In the present study, we investigated the effects of feeding WPC to mice on specific antibody responses to several orally or parenterally administered antigens, including influenza vaccine, diphtheria and tetanus toxoids, poliomyelitis vaccine, ovalbumin and cholera toxin sub-unit. WPC-fed mice produced elevated levels of antigen-specific intestinal tract and serum antibodies against all tested antigens, compared to mice that were fed a standard chow diet. Both primary and secondary intestinal tract antibody responses were elevated by WPC feeding, while only secondary serum responses were increased in WPC-fed mice. Significant up-regulation of intestinal tract antibody was observed within 2 weeks of primary oral immunizations. A period of pre-feeding with WPC, prior to commencement of immunization, did not alter the kinetics or magnitude of immune enhancement. These results identify bovine WPC as a potentially important dietary protein supplement, capable of enhancing humoral immune responses to a range of heterologous antigens. PMID- 12639818 TI - Gallium arsenide exposure impairs splenic B cell accessory function. AB - Gallium arsenide (GaAs) is utilized in industries for its semiconductor and optical properties. Chemical exposure of animals systemically suppresses several immune functions. The ability of splenic B cells to activate antigen-specific helper CD4(+) T cell hybridomas was assessed, and various aspects of antigen presenting cell function were examined. GaAs-exposed murine B cells were impaired in processing intact soluble protein antigens, and the defect was antigen dependent. In contrast, B cells after exposure competently presented peptides to the T cells, which do not require processing. Cell surface expression of major histocompatibility complex (MHC) class II molecules and several costimulatory molecules on splenic B cells, which are critical for helper T cell activation, was not affected by chemical exposure. GaAs exposure also did not influence the stability of MHC class II heterodimers, suggesting that the defect may precede peptide exchange. GaAs-exposed B cells contained a normal level of aspartyl cathepsin activity; however, proteolytic activities of thiol cathepsins B and L were approximately half the control levels. Furthermore, two cleavage fragments of invariant chain, a molecular chaperone of MHC class II molecules, were increased in GaAs-exposed B cells, indicative of defective degradation. Thus, diminished thiol proteolytic activity in B cells may be responsible for their impaired antigen processing and invariant chain degradation, which may contribute to systemic immunosuppression caused by GaAs exposure. PMID- 12639819 TI - OM197-MP-AC induces the maturation of human dendritic cells and promotes a primary T cell response. AB - Dendritic cell (DC) maturation is critical for the induction of antigen-specific T lymphocyte responses and may be essential for the development of human vaccines relying on T cell immunity. We investigated the effects on human DC of OM-197, a synthetic pseudodipeptide derived from amino acids, linked to three fatty acid chains and devoid of endotoxin properties. OM-197 upregulated the expression of HLA-DR, CD80, CD86, CD83, CD40 and CD54 at the surface of myeloid DC naturally present in blood as well as of DC generated in vitro from monocytes using IL-4 and GM-CSF. OM-197 also induced the release of IL-12 and TNF-alpha from DC. Finally, DC incubated with OM-197 after pulsing with hepatitis B surface antigen (HBs Ag) induced in vitro expansion of IFN-gamma-secreting HBs Ag-specific CD4(+) T lymphocytes from naive individuals. Taken together, these data identify OM-197 as a potential vaccine adjuvant for the induction of Th1-type responses. PMID- 12639820 TI - Cyclosporine A up-regulates expression of matrix metalloproteinase 2 and vascular endothelial growth factor in rat heart. AB - Cyclosporine A (CsA) is the first immunosuppressant universally used in allotransplantation. However, it has been demonstrated that this drug produces side effects in several organs, particularly in the kidney and in the heart. Since the immunosuppressive therapy induces myocardial toxicity, the aim of this study was to evaluate in the myocardium of CsA-treated rats the expression variations of vascular endothelial growth factor (VEGF) and matrix metalloproteinase 2 (MMP2), to verify if: VEGF increased, VEGF increase was associated with MMP2 increase and they could be considered as repair proteins. The study was carried out on 28 Wistar rats divided into two groups. The group I animals served as control and so they were subcutaneously injected daily with castor oil for 21 days; group II: animals were subcutaneously injected daily with CsA (Sandimmun, Sandoz) at the therapeutic dose (15 mg/kg) for 21 days. The group I animals (control) showed normal heart architecture and low levels of MMP2 and VEGF. The group II animals (CsA-treated) showed structural degenerative changes with myocardial fibrosis and a clear increase both in MMP2 and VEGF. These data show that the immunosuppressant therapy induces a high increase in both the proteins and that VEGF variations are associated with MMP2 variations. These findings suggest that these proteins are involved in the endogenous emergency cell mechanisms induced by stress conditions, probably playing a role against myocardial injury as repair proteins. PMID- 12639821 TI - Novel cyclic and linear oligopeptides that bind to integrin beta1 chain and either inhibit or costimulate T lymphocytes. AB - There is a redundancy of cellular beta1 integrin (very late antigen or VLA) receptors that mediate interactions between different extracellular matrix proteins (ECMP) and T lymphocytes. This suggests that antagonists targeted at individual VLA receptors may be of limited therapeutic efficacy in T cell mediated diseases and that agents such as monoclonal antibody 4B4, which bind to the common integrin beta1 chain and inhibit interactions between effector T cells and a range of ECMP, may be of greater therapeutic interest if toxicity can be avoided. We have therefore sought proof of principle as to whether small molecules that interact with the integrin beta1 chain at or near the 4B4 binding site can modulate T cell costimulation and adhesion in the presence of type I collagen or fibronectin (FN). Two phage display libraries, each expressing more than 10(9) independent cyclic or linear 7-mer peptides, were used to identify molecules of interest by an enrichment process involving specific recovery of phage bound to a human T cell line by elution with a large excess of 4B4 antibody. Novel cyclic and linear peptides have thus been identified and found to inhibit interactions between T cells and both type I collagen and fibronectin. A separate cyclic peptide was found to costimulate T cells in a beta1 integrin dependent manner. These findings form a basis for the development of small molecules that interact in inhibitory or stimulatory capacities with the common integrin beta1 chain, and may be of interest as therapeutic antagonists or immunologic adjuvants. PMID- 12639822 TI - Enhanced proliferation of functionally competent bone marrow cells in different strains of mice treated with swainsonine. AB - The immunomodulatory alkaloid swainsonine (8alphabeta-indolizidine 1alpha,2alpha,8beta-triol) has potential for overcoming the bone marrow suppressive effects of cancer chemotherapy and radiotherapy. An earlier study showed that multiple doses of swainsonine enhanced bone marrow cellularity in four different strains (C57BL/6; C3H-HEN; Balb/C and DBA-2 mice) of inbred mice which were not exposed to any chemotherapeutic agents or radiation. In vitro assessment of total colony formation capacity of bone marrow cells (BM CFUs) showed a 2- to 8-fold increase in swainsonine-treated mice compared to control mice that were given sham injections of physiological saline. In the current study, we have evaluated the functional competence of the bone marrow cells produced in response to swainsonine treatment of normal healthy mice. In particular, colony forming units-granulocyte-macrophage (CFU-GM), erythroid-burst forming units (BFUe) and CFU-Mix (or CFU-granulocyte-erythrocyte-monocyte megakaryocyte (CFU-GEMM)) levels, were determined using in vitro assays. The time course of the changes in CFU-GM, BFUe and CFU-Mix (CFU-GEMM) were also followed. Our results demonstrate that swainsonine bolsters the CFU capacity of BM cells without loss of function to levels which are several folds higher than in sham treated control mice. Swainsonine treatment caused an increase in all lineages of marrow cells without loss of function. This effect was reproduced in all four strains of inbred mice in this investigation. Examination of the peripheral blood did not reveal increase in white blood cells or changes in the hematocrit levels. The long-term effects of swainsonine treatment are not known at present. Nonetheless, swainsonine-induced increase in CFU capacity of bone marrow cells and related cells along the different differentiation paths should find clinical application in cancer treatment with chemotherapeutic agents and/or radiation. PMID- 12639823 TI - Obesity and oxidative stress: a direct link to CVD? PMID- 12639824 TI - Is oxidant stress a connection between obesity and atherosclerosis? PMID- 12639825 TI - Hyperhomocysteinemia: a million ways to lose control. PMID- 12639826 TI - Is there a need for cholesteryl ester transfer protein inhibition? PMID- 12639827 TI - Successful hunt for quantitative trait locus in thrombophilia. PMID- 12639828 TI - ATVB in focus: Platelet signal transduction and interactions with vascular cells. PMID- 12639829 TI - ATVB in focus: Smooth muscle cells. PMID- 12639831 TI - The beginning of the science underpinning occupational hygiene. AB - Sherwood and Greenhalgh's 1960 paper is a seminal one for the development of the science of human exposure. There are three key elements in the paper that deserve to be highlighted: the development of the first personal sampling pump and sampling head; the first comparison between personal sampling and static sampling; the first observation of the possible effect of personal sampling on the individual being sampled. PMID- 12639833 TI - Overview and characteristics of some occupational exposures and health risks on offshore oil and gas installations. AB - This review considers the nature, and recognition and control, of health risks in the offshore oil and gas industry from the occupational hygiene point of view. Particular attention is given to the changes in the nature of exposure and control of inhalation risks from substances hazardous to health in the UK sector, but other risks (e.g. dermatitis, noise and vibration) are also considered. The amount of published information on exposure to these hazards in the sector, or indeed on long-term health outcomes of working offshore, is limited. The approach taken to occupational health and hygiene in the sector has to be set in the context of the challenge of working in a remote and hostile environment where attention to safety and the need for emergency response to acute, rather than chronic, medical events are vital. However, changes in attitudes towards occupational health in the sector, legislation, the impact of environmental protection requirements and technology have all contributed to increasing the attention given to assessment and control of chemical and physical hazards. The health risks and benefits associated with the abandonment of installations, the application of new technologies, recovery of oil from ever deeper waters, lower staffing levels, environmental changes, the ageing workforce and the recognition of exposure patterns needing further attention/control (sequential multiple exposures, smaller workforce, peak/short-term exposures, etc.) are other current and future occupational hygiene challenges. PMID- 12639832 TI - Bioaerosol health effects and exposure assessment: progress and prospects. AB - Exposures to bioaerosols in the occupational environment are associated with a wide range of health effects with major public health impact, including infectious diseases, acute toxic effects, allergies and cancer. Respiratory symptoms and lung function impairment are the most widely studied and probably among the most important bioaerosol-associated health effects. In addition to these adverse health effects some protective effects of microbial exposure on atopy and atopic conditions has also been suggested. New industrial activities have emerged in recent years in which exposures to bioaerosols can be abundant, e.g. the waste recycling and composting industry, biotechnology industries producing highly purified enzymes and the detergent and food industries that make use of these enzymes. Dose-response relationships have not been established for most biological agents and knowledge about threshold values is sparse. Exposure limits are available for some contaminants, e.g. wood dust, subtilisins (bacterial enzymes) and flour dust. Exposure limits for bacterial endotoxin have been proposed. Risk assessment is seriously hampered by the lack of valid quantitative exposure assessment methods. Traditional culture methods to quantify microbial exposures have proven to be of limited use. Non-culture methods and assessment methods for microbial constituents [e.g. allergens, endotoxin, beta(1- >3)-glucans, fungal extracellular polysaccharides] appear more successful; however, experience with these methods is generally limited. Therefore, more research is needed to establish better exposure assessment tools and validate newly developed methods. Other important areas that require further research include: potential protective effects of microbial exposures on atopy and atopic diseases, inter-individual susceptibility for biological exposures, interactions of bioaerosols with non-biological agents and other potential health effects such as skin and neurological conditions and birth effects. PMID- 12639834 TI - Dust control measures in the construction industry. AB - Quartz is a human carcinogen and a causative agent of silicosis. Exposure levels often exceed exposure limits in the construction industry. The need for effective control measures is high, but the complex structure of the construction industry, the variability in sources of exposure and the frequent changes of worksite makes it difficult to implement even simple and potentially effective control measures. The aim of this study was to evaluate the impact of control measures for reducing quartz dust exposure and to assess the extent of their use. Full-shift respirable dust measurements (n = 61) and short-term measurements among construction workers were performed and results of a questionnaire study among 1335 construction workers were analysed. Full-shift measurements showed respirable quartz exposure levels up to 63 times the maximum allowable concentration (MAC) value (0.075 mg/m(3)). More than half of the measurements were above the MAC value. Control measures were not very strongly associated with the full-shift exposure estimates, but the short-term measurements showed large reduction factors (>70%) when wet dust suppression or local exhaust ventilation was used. The effectiveness of control measures is potentially high, and a significant part of the construction worker population is indeed using them on a regular basis. Still, both the exposure study and questionnaire survey show that the use of respiratory protection is the most widely used preventive measure in the construction industry. Respiratory protection might not always reduce exposure sufficiently. Only the combined use of more than one control measure can reduce exposures to acceptable levels. PMID- 12639836 TI - An in-field demonstration of the true relationship between skin infections and their sources in occupational diving systems in the North Sea. AB - INTRODUCTION: Skin infections in saturation diving are caused by microbes that flourish in saturation environments. Improvements in the prevention of infections must therefore be based on environmental control and elimination. Furthermore, only a few genotypes seem to be responsible for the majority of infections in the Norwegian sector of the North Sea, and these have all been demonstrated in saturation systems for many years. Although reservoirs of infectious genotypes have been identified, their true sources have not been identified. OBJECTIVES: The purpose of this field study was to log the contamination by Pseudomonas aeruginosa of the saturation system throughout a diving operation. MATERIALS AND METHODS: Daily water samples from the vessels drinking water system and from the heated seawater systems to divers suits were taken throughout the diving period of 1 month in the summer of 2001. All P.aeruginosa isolates were genotyped by pulsed field gel electrophoresis. RESULTS: A total of 17 P.aeruginosa genotypes were identified in the course of this field study. None of the most common infectious genotypes previously observed in the Norwegian sector were among these strains. Two genotypes were involved in skin infections during the period of operation: TP2 and TP12. TP2 was shown to be an inhabitant of the diving systems throughout the investigation period, while TP12 was introduced from seawater in the course of the operation and rapidly spread and established itself throughout the diving system. CONCLUSIONS: The study has demonstrated seawater as a true source of an infectious P.aeruginosa genotype in occupational diving systems. PMID- 12639835 TI - Exposure assessment to alpha- and beta-pinene, delta(3)-carene and wood dust in industrial production of wood pellets. AB - The main aim of the study was to measure the exposure to monoterpenes (alpha- and beta-pinene and Delta(3)-carene) and wood dust during industrial production of wood pellets and briquettes. Additional aims were to compare the results from wood dust sampled on a filter with real time measurements using a direct reading instrument and to identify peak exposures to dust. Twenty-four men working at six companies involved in industrial production of wood pellets and briquettes participated in the study. Monoterpenes were measured by diffusive sampling and wood dust was measured as total dust. A data logger (DataRAM) was used for continuous monitoring of dust concentration for 18 of the participants. The sampling time was approximately 8 h. The personal exposure to monoterpenes ranged from 0.64 to 28 mg/m(3) and a statistically significant (Kruskal-Wallis test, P = 0.0002) difference in levels of monoterpenes for workers at different companies was seen. In the companies the personal exposure to wood dust varied between 0.16 and 19 mg/m(3) and for 10 participants the levels exceeded the present Swedish occupational exposure limit (OEL) of 2 mg/m(3). The levels of wood dust during the morning shift were significantly (Mann-Whitney test, P = 0.04) higher compared with the afternoon shift. Continuous registration of dust concentration showed peak values for several working operations, especially cleaning of truck engines with compressed air. For 24 workers in six companies involved in industrial production of wood pellets the personal exposure to monoterpenes was low and to wood dust high compared with the present Swedish OEL and previous studies in Swedish wood industries. Since the DataRAM can identify critical working tasks with high wood dust exposure a reduction in exposure levels could probably be achieved by changes in working routines and by the use of protective equipment. PMID- 12639837 TI - Dust exposure during small-scale mining in Tanzania: a pilot study. AB - Small-scale mining in developing countries is generally labour-intensive and carried out with low levels of mechanization. In the Mererani area in the northern part of Tanzania, there are about 15000 underground miners who are constantly subjected to a poor working environment. Gemstones are found at depths down to 500 m. The objectives of this pilot study were to monitor the exposure to dust during work processes, which are typical of small-scale mining in developing countries, and to make a rough estimation of whether there is a risk of chronic pulmonary diseases for the workers. Personal sampling of respirable dust (n = 15) and 'total' dust (n = 5) was carried out during three consecutive days in one mine, which had a total of 50 workers in two shifts. Sampling started immediately before the miners entered the shaft, and lasted until they reappeared at the mine entrance after 5-8 h. The median crystalline silica content and the combustible content of the respirable dust samples were 14.2 and 5.5%, respectively. When drilling, blasting and shovelling were carried out, the exposure measurements showed high median levels of respirable dust (15.5 mg/m(3)), respirable crystalline silica (2.4 mg/m(3)), respirable combustible dust (1.5 mg/m(3)) and 'total' dust (28.4 mg/m(3)). When only shovelling and loading of sacks took place, the median exposures to respirable dust and respirable crystalline silica were 4.3 and 1.1 mg/m(3). This study shows that the exposure to respirable crystalline silica was high during underground small-scale mining. In the absence of personal protective equipment, the miners in the Mererani area are presumably at a high risk of developing chronic silicosis. PMID- 12639838 TI - Occupational health conditions in extreme environments. AB - The problems of work in extreme environments have been studied for many years. This paper discusses various aspects of work in hot and cold environments and at high pressure. PMID- 12639839 TI - Effect of CD14 blockade on endotoxin-induced acute lung injury in mice. AB - CD14 functions as a cell surface receptor for endotoxin (lipopolysaccharide [LPS]) and is thought to have an essential role in innate immune responses to infection. Previous studies have revealed attenuation of the systemic response after sepsis by blocking CD14. In this study, we tested the hypothesis that CD14 blockade protects against inflammatory responses associated with LPS pneumonia. We examined the effect of an anti-murine CD14 monoclonal antibody (4C1) on the development of acute lung injury induced by intratracheal LPS in mice. We also measured the production of cytokines (tumor necrosis factor-alpha, interleukin-6, and macrophage inflammatory protein-2) and nitric oxide by murine peritoneal macrophages exposed to LPS in vitro. Nuclear factor (NF)-kappa B translocation was evaluated in nuclear extracts from lung homogenates. 4C1 significantly attenuated pulmonary edema and neutrophil emigration after LPS administration. The production of cytokines and nitric oxide by LPS-stimulated macrophages was significantly decreased by 4C1 treatment. NF-kappa B translocation induced by LPS instillation was also suppressed by 4C1. These results suggest that blockade of CD14 might attenuate acute lung injury after intratracheal instillation of LPS through the suppression of NF-kappa B translocation. The inhibitory effect of CD14 blockade on cytokine production and nitric oxide release of macrophages might contribute to the attenuation of lung injury. PMID- 12639840 TI - Immunomodulatory effects of sensory nerves during respiratory syncytial virus infection in rats. AB - Respiratory syncytial virus (RSV) infection is associated with exaggerated neurogenic inflammation in the airways. This study sought to determine whether irritation of the mucosal sensory fibers affects the recruitment of lymphocytes and monocytes to RSV-infected airways. Pathogen-free rats were inoculated with RSV or with virus-free medium and were injected 5 days later with capsaicin to stimulate airway sensory nerves. Bronchoalveolar lavage was performed 1, 5, or 10 days after nerve stimulation, and samples were analyzed by differential cell count and flow cytometry. Without nerve stimulation, RSV caused a minimal increase in the number of lymphocytes and monocytes above pathogen-free control levels. After nerve stimulation, numerous lymphocytes, predominantly CD4+ T cells, and monocytes were recruited in the airways of infected rats, whereas no difference was found in pathogen-free controls. RSV induced overexpression of the neurokinin 1 (NK1) receptor for substance P on discrete lymphocyte subpopulations within the bronchial-associated lymphoid tissue (BALT), and treatment with a specific NK1 receptor antagonist abolished the recruitment of both lymphocytes and monocytes to infected airways. Our data suggest that airborne irritants stimulating mucosal sensory fibers during RSV infection exert important immunomodulatory effects by attracting to the infected airways selected lymphocyte subpopulations from the local BALT as well as monocytes. PMID- 12639841 TI - SP-B deficiency causes respiratory failure in adult mice. AB - Targeted deletion of the surfactant protein (SP)-B locus in mice causes lethal neonatal respiratory distress. To assess the importance of SP-B for postnatal lung function, compound transgenic mice were generated in which the mouse SP-B cDNA was conditionally expressed under control of exogenous doxycycline in SP-B-/ mice. Doxycycline-regulated expression of SP-B fully corrected lung function in compound SP-B-/- mice and protected mice from respiratory failure at birth. Withdrawal of doxycycline from adult compound SP-B-/- mice resulted in decreased alveolar content of SP-B, causing respiratory failure when SP-B concentration was reduced to <25% of normal levels. Decreased SP-B was associated with low alveolar content of phosphatidylglycerol, accumulation of misprocessed SP-C proprotein in the air spaces, increased protein content in bronchoalveolar lavage fluid, and altered surfactant activity in vitro. Consistent with surfactant dysfunction, hysteresis, maximal tidal volumes, and end expiratory volumes were decreased. Reduction of alveolar SP-B content causes surfactant dysfunction and respiratory failure, indicating that SP-B is required for postnatal lung function. PMID- 12639842 TI - Inhibition of prostaglandin synthesis during polystyrene microsphere-induced pulmonary embolism in the rat. AB - Our objective was to test the effect of inhibition of thromboxane synthase versus inhibition of cyclooxygenase (COX)-1/2 on pulmonary gas exchange and heart function during simulated pulmonary embolism (PE) in the rat. PE was induced in rats via intrajugular injection of polystyrene microspheres (25 micro m). Rats were randomized to one of three posttreatments: 1) placebo (saline), 2) thromboxane synthase inhibition (furegrelate sodium), or 3) COX-1/2 inhibition (ketorolac tromethamine). Control rats received no PE. Compared with controls, placebo rats had increased thromboxane B(2) (TxB(2)) in bronchoalveolar lavage fluid and increased urinary dinor TxB(2). Furegrelate and ketorolac treatments reduced TxB(2) and dinor TxB(2) to control levels or lower. Both treatments significantly decreased the alveolar dead space fraction, but neither treatment altered arterial oxygenation compared with placebo. Ketorolac increased in vivo mean arterial pressure and ex vivo left ventricular pressure (LVP) and right ventricular pressure (RVP). Furegrelate improved RVP but not LVP. Experimental PE increased lung and systemic production of TxB(2). Inhibition at the COX-1/2 enzyme was equally as effective as inhibition of thromboxane synthase at reducing alveolar dead space and improving heart function after PE. PMID- 12639843 TI - Magnitude-dependent regulation of pulmonary endothelial cell barrier function by cyclic stretch. AB - Ventilator-induced lung injury syndromes are characterized by profound increases in vascular leakiness and activation of inflammatory processes. To explore whether excessive cyclic stretch (CS) directly causes vascular barrier disruption or enhances endothelial cell sensitivity to edemagenic agents, human pulmonary artery endothelial cells (HPAEC) were exposed to physiologically (5% elongation) or pathologically (18% elongation) relevant levels of strain. CS produced rapid (10 min) increases in myosin light chain (MLC) phosphorylation, activation of p38 and extracellular signal-related kinase 1/2 MAP kinases, and actomyosin remodeling. Acute (15 min) and chronic (48 h) CS markedly enhanced thrombin induced MLC phosphorylation (2.1-fold and 3.2-fold for 15-min CS at 5 and 18% elongation and 2.1-fold and 3.1-fold for 48-h CS at 5 and 18% elongation, respectively). HPAEC preconditioned at 18% CS, but not at 5% CS, exhibited significantly enhanced thrombin-induced reduction in transendothelial electrical resistance but did not affect barrier protective effect of sphingosine-1 phosphate (0.5 microM). Finally, expression profiling analysis revealed a number of genes, including small GTPase rho, apoptosis mediator ZIP kinase, and proteinase activated receptor-2, to be regulated by CS in an amplitude-dependent manner. Thus our study demonstrates a critical role for the magnitude of CS in regulation of agonist-mediated pulmonary endothelial cell permeability and strongly suggests phenotypic regulation of HPAEC barrier properties by CS. PMID- 12639844 TI - Thickness of the blood-gas barrier in premature and 1-day-old newborn rabbit lungs. AB - The pulmonary capillaries of neonatal lungs are potentially vulnerable to stress failure because of the complex changes in the pulmonary circulation that occur at birth. We studied the ultrastructure of the blood-gas barrier (BGB) in premature and 1-day-old rabbit lungs and compared it with the ultrastructure of adult lungs. Normal gestation of rabbits is 30 days. After extensive pilot measurements, three premature (27 days gestation) and three newborn (1 day old) rabbit lungs were perfusion-fixed at arterial, venous, and airway pressures of 25, 0, and 10 cmH2O, respectively, and the measurements were compared with those of three adult lungs. The thickness of the capillary endothelium, alveolar epithelium, and interstitium of the BGB was measured at right angles to the barrier at random points. A striking finding was the large number of measurements of the interstitial thickness in 1-day-old lungs that were very thin (0-0.1 microm). The percentages of occurrence of very thin interstitium in premature, 1 day-old, and adult lungs were 35.3 +/- 9.4, 71.7 +/- 5.2, and 43.0 +/- 2.6, respectively (P < 0.02 for 1 day old vs. premature and adult). Given the previously found relationship between stress failure and interstitial thickness, this large proportion of very thin interstitial layers in the capillaries of 1 day-old lungs is a reasonable explanation for their previously demonstrated vulnerability to stress failure. PMID- 12639845 TI - Fibronectin matrix polymerization regulates small airway epithelial cell migration. AB - The continuous conversion of soluble fibronectin into extracellular matrix fibrils occurs through a dynamic, cell-dependent process. As the extracellular matrix is assembled, changes in the conformation of matrix proteins may expose biologically active, matricryptic sites that alter cell behavior. In this study, an in vitro model of wound healing was used to determine the role of matrix fibronectin in airway epithelial cell motility. Our findings indicate that, under basal conditions, small airway epithelial cell (SAEC) migration requires active fibronectin matrix polymerization. Furthermore, SAEC migration is increased significantly by the interaction of cells with a recombinant construct containing fibronectin's matricryptic III-1 site. In contrast, addition of increasing amounts of fibronectin to SAECs significantly decreased the rate of cell migration. This fibronectin-induced inhibition of cell migration was overcome by blocking excess fibronectin matrix deposition. These data indicate that SAEC migration is regulated in a biphasic manner by the polymerization of fibronectin in the extracellular matrix and suggest a stimulatory role for fibronectin's matricryptic III-1 site in cell motility. PMID- 12639846 TI - Thymulin evokes IL-6-C/EBPbeta regenerative repair and TNF-alpha silencing during endotoxin exposure in fetal lung explants. AB - Chorioamnionitis is associated with increased risks of perinatal respiratory failure; however, components of the inflammatory acute-phase response are known to actively promote lung maturation. To manipulate this relationship, we examined the effect of the thymic immunomodulator thymulin on fetal lung mesenchyme epithelial differentiation during exposure to Escherichia coli lipopolysaccharide (LPS). Gestation day 14 fetal rat lung explants were cultured for 96 h at fetal (23 mmHg) or ambient (142 mmHg) Po(2). Airway surface complexity (ASC, perimeter/ radical area(2)) was greater at fetal vs. ambient Po(2); however, exposure to 0.1 50 microg/ml LPS significantly raised ASC at 2 microg/ml in ambient Po(2) explants. LPS (50 microg/ml) depressed ASC in both conditions to untreated ambient Po(2) control values without changes in necrosis or apoptosis. To manipulate LPS-evoked TNF-alpha and IL-6 release, we exposed explants and A549 cells to combinations of 50 microg/ml LPS, 10 microM ZnCl(2), and 0.1-1,000 ng/ml thymulin at either Po(2). Thymulin+Zn(2+) suppressed and potentiated LPS-evoked TNF-alpha and IL-6 release, yielding an IC(50(TNF-alpha)) of 0.5 +/- 0.01 ng/ml and EC(50(IL-6)) of 1.4 +/- 0.3 ng/ml in A549 cells. This was accompanied by activation of the p38 MAPKMAPKAP-K2 pathway with sustained expression of TNF alpha and IL-6 transcripts at ambient Po(2). LPS+thymulin+Zn(2+)-treated explants showed proliferation of CCAAT-enhancer binding protein-beta (C/EBPbeta) and fibroblast growth factor-9 immunoreactive mesenchyme, which was abolished by IL-6 antisense oligonucleotides. The posttranscriptional suppression of immunogenic TNF-alpha synthesis coupled with raised IL-6 and C/EBPbeta-dependent mesenchyme proliferation suggests a role for bioactive thymulin in regulating regenerative repair in the fetal lung. PMID- 12639847 TI - Insulin does not mediate the attenuation of fatigue associated with glucose infusion in rat plantaris muscle. AB - Glucose infusion attenuates fatigue in rat plantaris muscle stimulated in situ, and this is associated with a better maintenance of electrical properties of the fiber membrane (Karelis AD, Peronnet F, and Gardiner PF. Exp Physiol 87: 585-592, 2002). The purpose of the present study was to test the hypothesis that elevated plasma insulin concentration due to glucose infusion ( approximately 900 pmol/l), rather than high plasma glucose concentration ( approximately 10-11 mmol/l), could be responsible for this phenomenon, because insulin has been shown to stimulate the Na+-K+ pump. The plantaris muscle was indirectly stimulated (50 Hz, for 200 ms, 5 V, every 2.7 s) via the sciatic nerve to perform concentric contractions for 60 min, while insulin (8 mU x kg-1x min-1: plasma insulin approximately 900 pmol/l) and glucose were infused to maintain plasma glucose concentration between 4 and 6 [6.2 +/- 0.4 mg x kg-1x min-1: hyperinsulinemic euglycemic (HE)] or 10 and 12 mmol/l [21.7 +/- 1.1 mg. kg-1. min-1: hyperinsulinemic-hyperglycemic clamps (HH)] (6 rats/group). The reduction in submaximal dynamic force was significantly (P < 0.05) less with HH (-53%) than with HE and saline only (-66 and -70%, respectively). M-wave characteristics were also better maintained in the HH than in HE and control groups. These results demonstrate that the increase in insulin concentration is not responsible for the increase in muscle performance observed after the elevation of circulating glucose. PMID- 12639849 TI - Kinetic analysis of pulmonary neutrophil retention in vivo using the multiple indicator-dilution technique. AB - Multiple-indicator-dilution experiments were performed in the lungs of 13 anesthetized dogs by simultaneous bolus injection of 111In-labeled neutrophils, 51Cr-labeled red blood cells, and Evans blue-labeled albumin. Concomitant counts of unlabeled neutrophils were similar in pulmonary artery and aortic blood samples, demonstrating a dynamic balance across the lungs in the physiological state. Outflow profiles of labeled neutrophils were analyzed on the basis of a recirculatory pharmacokinetic model of labeled albumin. The outflow profiles of the recovered neutrophils were composed of a throughput component of circulating neutrophils and a component of reversibly marginated neutrophils. They were interpreted by a model incorporating neutrophil margination (transfer coefficient = 0.195 +/- 0.081 s-1), rapid demargination (0.054 +/- 0.027 s-1), and transfer to a slow marginated pool (0.023 +/- 0.018 s-1). It will be interesting to apply the analysis in future studies aimed at determining whether it could be a useful research tool to investigate the interactions between the pulmonary endothelium and neutrophils in physiological and diseased states. PMID- 12639848 TI - Gravitational deposition in a rhythmically expanding and contracting alveolus. AB - In a previous simulation, our laboratory demonstrated that the flow induced by a rhythmically expanding and contracting alveolus is highly complex (Haber S, Butler JP, Brenner H, Emanuel I, and Tsuda A, J Fluid Mech 405: 243-268, 2000). Based on these earlier findings, we hypothesize that the trajectories and deposition of aerosols inside the alveoli differ substantially from those previously predicted. To test this hypothesis, trajectories of fine particles (0.5-2.5 microm in diameter) moving in the foregoing alveolar flow field and simultaneously subjected to the gravity field were simulated. The results show that alveolar wall motion is crucial in determining the enhancement of aerosol deposition inside the alveoli. In particular, 0.5- to 1-microm-diameter particles are sensitive to the detailed alveolar flow structure (e.g., recirculating flow), as they undergo gravity-induced convective mixing and deposition. Accordingly, deposition concentrations within each alveolus are nonuniform, with preferentially higher densities near the alveolar entrance ring, consistent with physiological observations. Deposition patterns along the acinar tree are also nonuniform, with higher deposition in the first half of the acinar generations. This is a result of the combined effects of enhanced alveolar deposition in the proximal region of the acinus due to alveoli expansion and contraction and reduction in the number of particles remaining in the gas phase down the acinar tree. We conclude that the cyclically expanding and contracting motion of alveoli plays an important role in determining gravitational deposition in the pulmonary acinus. PMID- 12639850 TI - Effect of voluntary facilitation on the diaphragmatic response to transcranial magnetic stimulation. AB - We assessed recruitment curves of the surface diaphragm motor-evoked potential (MEP) after transcranial magnetic stimulation during relaxation and at three different levels of facilitation (20, 40, and 60% of maximal inspiratory esophageal pressure) in 10 healthy subjects (six young and four elderly). MEP amplitude recruitment curves varied between individuals during relaxation and at each level of facilitation. Amplitude recruitment curves during relaxation were reproducible in individual subjects. Inspiratory maneuvers caused a decrease in motor threshold and latency and an increase in MEP amplitude, positively correlated to the intensity of facilitation. These changes were similar in young and elderly subjects. The best fit for MEP amplitude recruitment curves for each condition was obtained with a Boltzmann model. The performance of repeated submaximal inspiratory maneuvers did not affect the amplitude recruitment curves of the relaxed diaphragm. We conclude that the recruitment curve of the diaphragm with transcranial magnetic stimulation is repeatable and changes consistently with facilitation and will, therefore, be a robust experimental tool for the investigation of supraspinal pathways to the diaphragm. PMID- 12639852 TI - Heat balance precedes stabilization of body temperatures during cold water immersion. AB - Certain previous studies suggest, as hypothesized herein, that heat balance (i.e., when heat loss is matched by heat production) is attained before stabilization of body temperatures during cold exposure. This phenomenon is explained through a theoretical analysis of heat distribution in the body applied to an experiment involving cold water immersion. Six healthy and fit men (mean +/ SD of age = 37.5 +/- 6.5 yr, height = 1.79 +/- 0.07 m, mass = 81.8 +/- 9.5 kg, body fat = 17.3 +/- 4.2%, maximal O2 uptake = 46.9 +/- 5.5 l/min) were immersed in water ranging from 16.4 to 24.1 degrees C for up to 10 h. Core temperature (Tco) underwent an insignificant transient rise during the first hour of immersion, then declined steadily for several hours, although no subject's Tco reached 35 degrees C. Despite the continued decrease in Tco, shivering had reached a steady state of approximately 2 x resting metabolism. Heat debt peaked at 932 +/- 334 kJ after 2 h of immersion, indicating the attainment of heat balance, but unexpectedly proceeded to decline at approximately 48 kJ/h, indicating a recovery of mean body temperature. These observations were rationalized by introducing a third compartment of the body, comprising fat, connective tissue, muscle, and bone, between the core (viscera and vessels) and skin. Temperature change in this "mid region" can account for the incongruity between the body's heat debt and the changes in only the core and skin temperatures. The mid region temperature decreased by 3.7 +/- 1.1 degrees C at maximal heat debt and increased slowly thereafter. The reversal in heat debt might help explain why shivering drive failed to respond to a continued decrease in Tco, as shivering drive might be modulated by changes in body heat content. PMID- 12639853 TI - Coronary flow: clinical considerations. AB - In the measurement of coronary blood flow to determine the success of percutaneous coronary intervention, invasive techniques, coupled with plaque characterisation and other intracoronary imaging modalities, may prove invaluable. PMID- 12639854 TI - Computed tomography imaging of cardiac tamponade secondary to a posterior pericardial abscess. PMID- 12639855 TI - An unusual presentation of delayed cardiac perforation caused by atrial screw-in lead. PMID- 12639856 TI - Cardiac surgery and the brain: differences between adult and paediatric studies. AB - Evidence is growing that patients with congenital heart disease who undergo surgery may be at increased risk of neurodevelopmental dysfunctions, particular paediatric survivors. However, paediatric studies involve different challenges from those conducted on adults. PMID- 12639857 TI - Protecting the damaged heart during coronary surgery. AB - In coronary surgery, protective strategies that offer adequate safety for patients with healthy ventricles may not be adequate for those with severe impairment of ventricular function. PMID- 12639858 TI - Pulmonary thromboendarterectomy almost 50 years after the first surgical attempts. AB - With increased experience in preoperative evaluation, surgical approach and postoperative care of chronic thromboembolic pulmonary hypertension, pulmonary thromboendarterectomy can be performed with an acceptably low risk of death. Most patients, even those in a very compromised state, have excellent, long lasting results. PMID- 12639860 TI - Prevention of embolisation during percutaneous vein graft intervention using a Filter Wire distal protection device. PMID- 12639862 TI - Prolonged cardiac arrest caused by glossopharyngeal neuralgia. PMID- 12639861 TI - Long QTc predicts future cardiac death in stroke survivors. AB - OBJECTIVES: To test the hypothesis that the QTc of any lead of the ECG predicts death after stroke, and to determine which lead of the ECG carries the greatest risk of cardiac death when its QTc is measured. DESIGN: Standard 12 lead ECGs were analysed by one observer who was blind to patient outcome. SETTING: A major teaching hospital in Scotland. PATIENTS: 404 stroke survivors were studied at approximately one year after the cerebrovascular event and followed for up to 6.3 years. OUTCOME MEASURES: Death from any cause and cardiac mortality. RESULTS: The QTc measured from any lead of the ECG (except aVR) was associated with death from any cause. A prolonged QTc in limb lead III and chest lead V6 carried the highest relative risk of cardiac death (a 3.1-fold incease). After adjusting for overt ischaemic heart disease, pulse pressure, glucose, and cholesterol, a prolonged QTc in lead V6 was associated with a relative risk of cardiac death of 2.8 (95% confidence interval (CI) 1.1 to 7.3) (p = 0.028) and of death from all causes of 2.9 (95% CI 1.6 to 5.3) (p < 0.001). If the QTc in V6 exceeded 480 ms, then the specificity of predicting cardiac death within five years after the stroke was 94%. CONCLUSIONS: Although treatment of the conventional modifiable risk factors is important, stroke survivors with a prolonged QTc in lead V6 are still at a high risk of cardiac death and may need more intensive investigations and treatments than are currently routine practice. PMID- 12639859 TI - The therapeutic potential of new insights into myocardial excitation-contraction coupling. AB - The physiological mechanisms that link myocyte depolarisation and contraction are referred to collectively as excitation-contraction coupling. This important process uses calcium as a second messenger to convert electrical depolarisation of the myocyte sarcolemma into the coordinated contraction of the cell's internal myofilament apparatus. The inotropic properties of the cell are determined by the efficiency of this process and when this efficiency is lost contractile dysfunction and heart failure develop, along with a propensity for arrhythmias. Previous attempts to use positive inotropic drugs in the management of chronic heart failure have been disappointing. Such drugs have been associated with unacceptable side effects and worse morbidity and mortality outcomes, primarily through their non-specific amplification of intracellular cascade pathways that modify the cell's inotropic state. As a result of recent advances in our understanding of how excitation-contraction coupling works in both health and disease it may be possible to design more specifically targeted drug treatment that has the potential to avoid the detrimental effect of currently available drugs while at the same time improving the inotropic properties of the cell. PMID- 12639864 TI - Patency of coronary artery lumen surrounded by metallic stent evaluated by three dimensional volume rendering images using ECG gated multislice computed tomography. PMID- 12639863 TI - Persistence of systolic coronary flow reversal predicts irreversible dysfunction after reperfused anterior myocardial infarction. AB - OBJECTIVE: To investigate serial assessments of systolic coronary flow reversal in the infarct related artery for predicting poor left ventricular functional recovery after reperfused acute myocardial infarction. SETTING: Regional hospital. PATIENTS AND METHODS: 49 patients with anterior acute myocardial infarction had transthoracic Doppler echocardiography to record coronary flow velocity in the left anterior descending coronary artery immediately after successful primary coronary angioplasty (day 0), and at 48 hours, one week, and three weeks. MAIN OUTCOME MEASURES: Coronary flow velocity at each time point; regional wall motion score index (RWMSI) at day 0 and at three weeks. Irreversible dysfunction was defined as a decrease in RWMSI to < 0.22. RESULTS: Measurements of coronary flow velocity could be made in 45 patients. Patients were divided into three groups: no systolic flow reversal (group 1, n = 27), systolic flow reversal observed only on day 0 (group 2, n = 8), and systolic flow reversal persisting until 48 hours (group 3, n = 10). Although baseline RWMSI was similar among the three groups, the value at three weeks was significantly higher in group 3 than in the other two groups. In predicting irreversible dysfunction, the persistence of systolic flow reversal up to 48 hours had a higher positive predictive value (100%) than the presence of systolic flow reversal on day 0 (67%, p < 0.04). The negative predictive value of systolic flow reversal at 48 hours (83%) was comparable in accuracy to the presence of systolic flow reversal on day 0 (85%, NS). CONCLUSIONS: In reperfused anterior acute myocardial infarction, serial assessment of coronary flow velocity in the left anterior descending coronary artery is feasible using transthoracic Doppler echocardiography, and the persistence of systolic flow reversal at 48 hours is a more specific marker of irreversible dysfunction than peak creatine kinase or diastolic deceleration time. PMID- 12639865 TI - White blood cell count and long term mortality after non-ST elevation acute coronary syndrome treated with very early revascularisation. AB - OBJECTIVE: To evaluate the predictive value of white blood cell count (WBC) for short and long term mortality in patients with non-ST elevation acute coronary syndromes (NSTACS) treated with a very early invasive strategy. DESIGN: Prospective cohort study in 1391 consecutive patients with NSTACS undergoing very early revascularisation. Patients were stratified according to quartiles of WBC determined on admission. RESULTS: Kaplan-Meier survival analysis showed a cumulative three year survival of 93.8% in the first quartile of WBC (< 6800/mm(3)), 94.4% in the second quartile (6800-8000/mm(3)), 95.1% in the third quartile (8000-10000/mm(3)), and 82.4% in the fourth quartile (> 10000/mm(3)) at 36 months (p < 0.001 by log rank). Relative to patients in the three lower WBC quartiles, patients in the highest quartile were three times more likely to die during the hospitalisation (hazard ratio 3.2, 95% confidence interval (CI) 1.5 to 7.1; p = 0.003) and during long term follow up (hazard ratio 3.4, 95% CI 2.2 to 5.3; p < 0.001). By multivariate Cox regression analysis including baseline demographic, clinical, and angiographic covariables, WBC in the highest quartile remained a strong independent predictor of mortality (hazard ratio 3.3, 95% CI 1.9 to 5.6; p < 0.001). CONCLUSIONS: WBC is a strong independent predictor of short and long term mortality after NSTACS treated with very early revascularisation. PMID- 12639866 TI - Impact of microvascular integrity and local viability on left ventricular remodelling after reperfused acute myocardial infarction. AB - OBJECTIVE: To assess left ventricular remodelling in patients with reperfused acute myocardial infarction and to study its relation to microvascular damage. PATIENTS: 25 patients successfully treated by primary percutaneous coronary angioplasty for acute myocardial infarction. SETTING: University hospital METHODS: Indexed end diastolic (EDVi) and end systolic (ESVi) volumes were assessed on admission and repeated at days 1 and 8. Coronary flow reserve (CFR) was assessed in the infarct related artery on day 1. Myocardial blood volume was assessed on admission and at day 8 by myocardial contrast echocardiography. In patients who manifested persistent myocardial dysfunction at hospital discharge (n = 21), local inotropic reserve was assessed by dobutamine echocardiography at day 7. RESULTS: On admission, patients with and without local viability had similar EDVi and ESVi (EDVi 67 (9) and 73 (14) ml/m(2), respectively; ESVi 34 (8) and 40 (11) ml/m(2), respectively; NS). EDVi increased to 97 (22) ml/m(2) in patients without local viability (p < 0.01 v admission) but remained unchanged at 70 (11) ml/m(2) in patients with viable myocardium (NS v admission). For pooled patient data, the percentage change in EDVi correlated with CFR (r = 0.76, p < 0.0001) and myocardial blood volume in the infarct territory (r = 0.80, p < 0.0001). CONCLUSION: Left ventricular dilatation may preferably occur in patients without local viability and is correlated with early CFR and extent of myocardial blood volume in the infarct territory. PMID- 12639867 TI - Variation during the week in the incidence of acute myocardial infarction: increased risk for Japanese women on Saturdays. AB - BACKGROUND: Variations in the incidence of acute myocardial infarction during the week may differ between and within communities, according to lifestyle. OBJECTIVE: To identify potential triggering factors for acute myocardial infarction by examining variations in incidence in the days of the week within the Osaka area of Japan. PATIENTS: Of 2511 consecutive patients in this region who were admitted to hospital for acute myocardial infarction between April 1998 and March 2001 and consented to take part, 2400 who had a definitely identified time of onset were enrolled. RESULTS: For this group as a whole, no significant difference in incidence was noted between days of the week. However, in subgroup analyses women were shown to have significant variation through the week, peaking on Saturday with a 39% increase in relative risk (p = 0.037); working subjects showed a peak on Monday, with a 26% increase in relative risk (p = 0.038). Stratified analyses showed that in working men there was a prominent Monday peak in the onset of infarction, with a 30% increase in relative risk (p = 0.022), while in working women, there was no significant variation through the week. CONCLUSIONS: Earlier findings of a Monday peak linked to increased physical and mental occupational stress are confirmed. There is also an increase in uncertain risk factors on Saturdays for Japanese women, possibly involving a stressful weekend burden for women. Confirmation of this finding in other communities may help identify triggers of acute myocardial infarction and be useful in prevention. PMID- 12639868 TI - Attenuation of hypercapnic carbon dioxide chemosensitivity after postinfarction exercise training: possible contribution to the improvement in exercise hyperventilation. AB - OBJECTIVE: To elucidate the responsible mechanisms of increased slope of minute ventilation relative to carbon dioxide production (VE/VCO(2)) during exercise after acute myocardial infarction without overt signs of heart failure, patients who had an acute myocardial infarction were examined after participating in a three month supervised exercise training programme. DESIGN: Exercise testing, hypercapnic CO(2) chemosensitivity measurement (rebreathing method), and pulmonary function test were repeated at entry and after three months in 50 acute myocardial infarction patients with neither symptoms nor signs of heart failure who completed the training programme. Ten patients who performed initial inhospital training served as controls. RESULTS: Age, peak oxygen uptake, left ventricular ejection fraction, CO(2) chemosensitivity, respiratory parameters (percentage of predicted normal vital capacity (%VC), forced expiratory volume in one second, and carbon monoxide transfer factor (%TLCO)) were all significantly correlated with VE/VCO(2) slope. Multivariate regression analysis showed that age (beta = 0.29, p = 0.01), %TLCO (beta = -0.27, p = 0.01), and CO(2) chemosensitivity (beta = 0.49, p < 0.001) were independent determinants of VE/VCO(2) slope. After three months, there was no significant change in these parameters in the control group. Peak oxygen uptake, %TLCO, and %VC and attenuation in CO(2) chemosensitivity increased significantly in the training group. The VE/VCO(2) slope decreased marginally (p = 0.11). The changes in VE/VCO(2) slope were correlated only with those in CO(2) chemosensitivity (r = 0.50, p < 0.001). CONCLUSION: After acute myocardial infarction, exercise hyperventilation is seen in association with aging, enhanced hypercapnic CO(2) chemosensitivity, and reduced TLCO, even in the absence of overt heart failure. The correlation of VE/VCO(2) attenuation after training with the reduction in CO(2) chemosensitivity suggests that exercise training may reduce increased VE/VCO(2) slope, at least partially by reducing CO(2) chemosensitivity. PMID- 12639870 TI - Evolution of statin prescribing 1994-2001: a case of agism but not of sexism? AB - OBJECTIVE: To study trends in the use of lipid lowering drugs in the UK, and to assess which patient factors influence prescribing. METHODS: Routinely collected computerised medical data were analysed from 142 general practices across England and Wales that provide data for the Doctors' Independent Network database. Subjects included were people aged 35 years or more with treated ischaemic heart disease, averaging annually over 30,000. The temporal trend from 1994 to 2001 in prescription of lipid lowering drugs and daily statin dose and the odds ratios (ORs) for receiving a statin prescription in 1998 were examined. RESULTS: Lipid lowering drug prescribing increased greatly over time, entirely because of statins, so that in 2001 56.3% of men and 41.1% of women with ischaemic heart disease received lipid lowering drugs. However, 33% of these patients were on a < 20 mg simvastatin daily equivalent. In 1998 the OR for receiving a statin fell from 1 at age 55-64 to 0.64 at 65-74 and 0.16 at 75-84 years. The age effect was similar in those without major comorbidity. Revascularised patients were much more likely to receive a statin than those with angina (OR 3.92, 95% confidence interval (CI) 3.57 to 4.31). Men were more likely to receive a statin than women (OR 1.62, 95% CI 1.54 to 1.71) but this difference disappeared after adjustment for age and severity of disease (OR 1.06). Geographical region had little effect but there was a very weak socioeconomic gradient. CONCLUSIONS: Although prescribing has increased, many patients who may benefit from lipid lowering drugs either do not receive it or are undertreated, possibly because of lack of awareness of the relative potency of the different statins. Patients with angina and the elderly are less likely to receive treatment that may prevent a coronary event. PMID- 12639869 TI - Serum magnesium aberrations in furosemide (frusemide) treated patients with congestive heart failure: pathophysiological correlates and prognostic evaluation. AB - OBJECTIVES: To determine the prevalence of hypomagnesaemia and hypermagnesaemia, to discern various factors associated with abnormal serum magnesium, and to estimate prognostic significance of serum magnesium aberrations in patients with congestive heart failure. DESIGN: Observational study. SETTING: Medical department of a university hospital (tertiary referral centre). PATIENTS: 404 consecutive patients admitted with congestive heart failure as one of the diagnoses and previously treated with furosemide (frusemide) for at least three months. MAIN OUTCOME MEASURES: Clinical, biochemical, and electrocardiographic variables were analysed with respect to serum magnesium aberrations. Following discharge, mortality rates, including sudden death, were registered. RESULTS: Hypomagnesaemia was found in 50 patients (12.3%) and 20 (4.9%) were hypermagnesaemic. Female sex (p < 0.04), diabetes mellitus (p < 0.006), hypocalcaemia (p = 0.03), hyponatraemia (p < 0.05), malignant disease (p = 0.05), and high fever (p = 0.05) were statistically associated with hypomagnesaemia. Renal failure, severe congestive heart failure, and high dose furosemide treatment (> 80 mg/day) were associated with hypermagnesaemia (p < 0.001, p = 0.05, and p < 0.03, respectively). Hypermagnesaemic patients were older and weighed less. On follow up (median duration 43 months), 169 (41.8%) died, with 22 (13%) sudden deaths. Mortality was highest with hypermagnesaemia, lowest with normomagnesaemia, and intermediate with hypomagnesaemia. After adjustment for renal failure, old age, and severity of congestive heart failure, hypomagnesaemia but not hypermagnesaemia emerged as being significantly associated with shorter survival (p = 0.009). No statistical association was found between sudden death and magnesium concentrations. CONCLUSIONS: While hypermagnesaemia seems to represent a prognostic marker only, hypomagnesaemia appears to have an adverse pathophysiological effect. The subgroup of patients at risk for hypomagnesaemia requires frequent serum magnesium determinations and magnesium replacement for as long as hypomagnesaemia persists. PMID- 12639871 TI - Arterial dysfunction in syndrome X: results of arterial reactivity and pulse wave propagation tests. AB - OBJECTIVE: To assess arterial distensibility using pulse wave velocity (PWV) measurements and its relation with endothelium dependent vasodilatation (EDV) in patients with cardiological syndrome X. METHODS: The study group consisted of 92 patients: 52 with syndrome X (34 women, 18 men, mean (SD) age 45 (3) years) and 40 healthy volunteer controls (27 women, 13 men, mean (SD) age 41 (2) years) without risk factors of atherosclerosis and with negative ECG exercise test and normal proximal coronaries on transoesophageal echocardiography. Patients with arterial hypertension, diabetes mellitus, valvar disease, or cardiomyopathy were excluded. PWV measured by a Complior Colson device was calculated for each patient. EDV was assessed from two dimensional Doppler measurement using an Acuson Sequoia with 8 MHz linear transducer at rest, during postischaemic reactive hyperaemia, and after an oral dose of 400 micro g of glyceryl trinitrate. RESULTS: PWV was significantly higher in patients with syndrome X than in healthy subjects (9.3 (0.7) m/s v 8.2 (0.9) m/s, respectively, p < 0.001). Baseline brachial artery diameter was similar in the syndrome X and control groups (4.0 (0.6) mm v 4.08 (0.64) mm, NS). EDV was impaired in patients with syndrome X compared with controls (6.6 (3.0)% v 11.1 (3.9)%, p < 0.001). Endothelium independent vasodilatation was similar in both groups. In patients with syndrome X there was a positive correlation between PWV and the degree of EDV (r = 0.864, p < 0.001). The cut off value for PWV was 8.5 m/s, with a sensitivity of 62% and a specificity of 91%. CONCLUSIONS: EDV but not glyceryl trinitrate induced vasodilatation is decreased in patients with syndrome X. There is a strong correlation between PWV and the degree of endothelial dysfunction of peripheral arteries in patients with syndrome X. PWV assessment may be useful to identify abnormal vascular physiology in these patients. PMID- 12639873 TI - Coronary surgery with non-cardioplegic methods in patients with advanced left ventricular dysfunction: immediate and long term results. AB - OBJECTIVE: To evaluate perioperative results and long term survival in patients with severe left ventricular (LV) dysfunction undergoing coronary artery bypass grafting (CABG) using non-cardioplegic methods. METHODS: From April 1990 through December 1999, 4100 consecutive patients underwent isolated CABG using hypothermic ventricular fibrillation. Of these, 141 (3.4%) had severe LV dysfunction (ejection fraction < 30%). Mean age was 58.3 (9.6) years. 64 patients (45.4%) were in Canadian Cardiovascular Society class III or IV and 16 (11.3%) were subjected to urgent or emergent surgery. A previous myocardial infarction was recorded in 127 (90.1%). The majority (89.4%) had triple vessel and 26 (18.4%) had left main disease. The mean number of grafts per patient was 3.1. At least one internal thoracic artery was used in all patients and 21 (14.8%) had bilateral internal thoracic artery grafts (1.2 arterial grafts per patient). RESULTS: Perioperative mortality was 2.8% (4 patients) and the incidence of acute myocardial infarction 2.8%. 50 (35.5%) patients required inotropes but only 16 (11.3%) required it for longer than 24 hours; 5 patients (3.5%) needed mechanical support. The incidence of renal failure was 3.5%. Mean duration of hospital stay was 9.6 (8.3) days. Follow up was 95% complete and extended for a mean of 57 (30) months. Late mortality was 11.5%. Actuarial survival rates at 1, 3, and 5 years were 96%, 91%, and 86%, respectively. CONCLUSIONS: Non-cardioplegic techniques are safe and effective in preserving the myocardium during CABG in patients with coronary artery disease and poor LV function, with low operative mortality and morbidity, and encouraging medium to long term survival rates. PMID- 12639875 TI - Validation of four different risk stratification systems in patients undergoing off-pump coronary artery bypass surgery: a UK multicentre analysis of 2223 patients. AB - BACKGROUND: Various risk stratification systems have been developed in coronary artery bypass graft surgery (CABG), based mainly on patients undergoing procedures with cardiopulmonary bypass. OBJECTIVE: To assess the validity and applicability of the Parsonnet score, the EuroSCORE, the American College of Cardiology/American Heart Association (ACC/AHA) system, and the UK CABG Bayes model in patients undergoing off-pump coronary artery bypass surgery (OPCAB) in the UK. METHODS: Data on 2223 patients who underwent OPCAB in eight cardiac surgical centres were collected. Predicted mortality risk scores were calculated using the four systems and compared with observed mortality. Calibration was assessed by the Hosmer-Lemeshow (HL) test. Discrimination was assessed using the receiver operating characteristic (ROC) curve area. RESULTS: 30 of 2223 patients (1.3%) died in hospital. For the Parsonnet score the HL test was significant (p < 0.001) and the receiver operating characteristic curve (ROC) area was 0.74. For the EuroSCORE the HL test was also significant (p = 0.008) and the ROC area was 0.75. For the ACC/AHA system the HL test was non-significant (p = 0.7) and the ROC area was 0.75. For the UK CABG Bayes model the HL test was also non significant (p = 0.3) and the ROC area was 0.81. CONCLUSIONS: The UK CABG Bayes model is reasonably well calibrated and provides good discrimination when applied to OPCAB patients in the UK. Among the other three systems, the ACC/AHA system is well calibrated but its discrimination power was less than for the UK CABG Bayes model. These data suggest that the UK CABG Bayes model could be an appropriate risk stratification system to use for patients undergoing OPCAB in the UK. PMID- 12639877 TI - Obstacles to the initiation of beta blockers for heart failure in a specialised clinic within a district general hospital. PMID- 12639876 TI - Neurodevelopment at 1 year of age in infants with congenital heart disease. AB - OBJECTIVE: To assess psychomotor development and neurological sequelae in infants after surgery for congenital heart defects. DESIGN AND SETTING: Single institution prospective cohort study. PATIENTS: 90 of 112 consecutive surviving infants of less than 1 year of age, without brain anomalies, conditions, or syndromes associated with delayed mental development, who underwent cardiac surgery during an 18 month period; 20 control infants with minor or no congenital heart defects. MAIN OUTCOME MEASURES: Griffiths developmental scales and standardised neurological examination at 1 year. RESULTS: Mean (SD) developmental quotient (DQ) in index infants was 99 (10.6), compared with 106.7 (6.6) in controls (p < 0.001). DQ was lower in infants after palliative surgery (n = 16; 88 (12.2)) than after corrective surgery (n = 74; 101.4 (8.6)) (p < 0.001). Of the 90 index infants, 24 (27%) had a DQ below 93.5 (more than 2 SD below the mean of controls). Developmental delay (DQ < 93.5) was more common after palliative surgery (10/16, 63%) than after corrective surgery (14/74, 19%) (p < 0.001). Of the 90 index infants, 29 (32%) had neurological abnormalities, compared with only one of the 20 controls (5%) (p = 0.013). Neurological abnormalities were more frequent after palliative surgery (11/16, 69%) than after corrective surgery (18/74, 24%) (p < 0.001). CONCLUSIONS: There is a considerable rate of neurodevelopmental impairment at 1 year of age in infants after cardiac surgery. Psychomotor impairment and neurological sequelae are apparently more severe in infants in whom only palliative surgery is possible. PMID- 12639879 TI - Lack of association between angiotensin converting enzyme (ACE) genotype, serum ACE activity, and haemodynamics in patients with primary pulmonary hypertension. PMID- 12639878 TI - Three dimensional images with ECG gated multislice computed tomography revealed stenosis of the descending aorta in 2 month old baby. PMID- 12639880 TI - Position and Mobilisation Post-Angiography Study (PAMPAS): a comparison of 4.5 hours and 2.5 hours bed rest. PMID- 12639881 TI - An unusual complication of transvenous temporary pacing. PMID- 12639882 TI - Conduit arterial stiffness is associated with impaired left ventricular subendocardial function. PMID- 12639883 TI - Lipid lowering to delay the progression of coronary artery disease. PMID- 12639884 TI - Management of ischaemic mitral regurgitation. PMID- 12639886 TI - Neonatal ECG changes caused by supratherapeutic flecainide following treatment for fetal supraventricular tachycardia. PMID- 12639885 TI - Challenges in the care of adult patients with congenital heart defects. PMID- 12639887 TI - A diagnostic pitfall in a patient with an implantable cardioverter-defibrillator. PMID- 12639889 TI - Successful radiofrequency catheter ablation of "clockwise" and "counterclockwise" bundle branch re-entrant ventricular tachycardia in the absence of myocardial or valvar dysfunction without detecting bundle branch potentials. AB - A case is reported of a patient with only isolated conduction abnormalities of the His-Purkinje system with no identifiable myocardial or valvar dysfunction, leading to "clockwise" and "counterclockwise" bundle branch re-entrant ventricular tachycardias (BBRVTs). The electrophysiological study showed infra Hisian conduction system disease and two different inducible wide QRS complex tachycardias. Neither right bundle branch nor left bundle branch potentials were recorded despite extensive catheter manipulation. However, these tachycardias were diagnosed as BBRVTs by using entrainment manoeuvres and comparing the HV intervals during both sinus rhythm and the tachycardias. These tachycardias were eliminated by catheter ablation of the right bundle branch, using the morphology of the local electrograms and anatomical findings. PMID- 12639890 TI - Concordance for hypoplastic left heart syndrome in a monochorionic twin pregnancy. AB - The risk of structural heart disease is significantly higher in twin pregnancies than in singleton pregnancies, but the concordance rate has been found to be relatively low, even in monochorionic pregnancies. This is the first report of a monochorionic twin pregnancy concordant for hypoplastic left heart syndrome (HLHS), the diagnosis having been made by fetal echocardiography at 15 weeks' gestation. The findings were confirmed at necropsy at 17 weeks' gestation, following termination of pregnancy. Both twins had mitral and aortic atresia, with severely hypoplastic aortic arches. This report adds weight to there being a genetic component to the cause of HLHS in some cases and illustrates how the findings from early fetal echocardiography with postmortem follow up can help to extend the understanding of the aetiology of this condition. PMID- 12639891 TI - Toad venom poisoning: resemblance to digoxin toxicity and therapeutic implications. AB - A healthy man developed gastrointestinal symptoms after ingesting purported aphrodisiac pills. He had severe unrelenting bradycardia, hyperkalaemia, and acidosis. He rapidly developed severe life threatening cardiac arrhythmias and died after a few hours. He was found to have positive serum digoxin concentrations, although he was not taking digoxin. Toad venom poisoning is similar to digitalis toxicity and carries a high mortality. Cardiac glycoside poisoning can occur from ingestion of various plants and animal toxins, and the venom gland of cane toad (Bufo marinus) contains large quantities of cardiac glycosides. Toad venom, a constituent of an aphrodisiac, was considered responsible for the development of clinical manifestations and death in this patient. Digoxin specific Fab fragment has been reported to be beneficial in the treatment of toad venom poisoning. This report alerts physicians to the need to be aware of a new community toxic exposure, as prompt treatment with digoxin specific Fab fragment may be life saving. The treatment approach to patients with suspected toad venom poisoning is described. PMID- 12639892 TI - Administration of PTH-(7-84) antagonizes the effects of PTH-(1-84) on bone in rats with moderate renal failure. AB - Circulating parathyroid hormone (PTH) is a mixture of PTH-1-84 and carboxy terminal (C-PTH) fragments. Recently, the "intact" PTH assay was reported to detect not only PTH-(1-84) but also a C-PTH fragment, presumably PTH-(7-84). The purpose of this study was to determine whether PTH-(7-84) antagonizes the PTH-(1 84) effects on bone. Forty-eight rats were thyroparathyroidectomized (TPTX), eight were used as controls and the remaining TPTX rats (10/group) were nephrectomized (Nx) and subsequently given PTH-(1-84), PTH-(7-84), PTH-(1-84) and PTH-(7-84) or no PTH peptide. Another ten rats were sham-operated and served as controls. Administration of PTH-(1-84) brought serum calcium and osteoblast activity i.e., bone turnover, to normal levels. Concomitant administration of PTH (7-84) abrogated these effects. Administration of PTH-(7-84) alone did not further suppress the levels of serum calcium and bone turnover in these TPTX-Nx rats. Moreover, radioligand binding experiments demonstrate that PTH-(7-84) binds specifically to SaOS cells and is equally displaced by both PTH-(1-84) and (7 84), but only partially displaced by PTH-(1-34). These data indicate that PTH-(7 84) antagonizes PTH-(1-84) effect not only on serum calcium but also on osteoblasts to affect bone turnover. PMID- 12639893 TI - TGF-beta3 regulates the blood-testis barrier dynamics via the p38 mitogen activated protein (MAP) kinase pathway: an in vivo study. AB - Recent studies using Sertoli cells cultured in vitro to permit tight junction (TJ) assembly have shown that TJ dynamics are regulated, at least in part, by TGF beta3 via the p38 mitogen activated protein (MAP) kinase pathway. This in turn regulates the production of occludin, a TJ-integral membrane protein, by Sertoli cells. Yet it is not known if this pathways is used by Sertoli cells to regulate the blood-testis barrier (BTB) function in vivo. Using an in vivo model for studying BTB dynamics, we report herein the CdCl(2)-induced BTB damage in rats was associated with a significant reduction in testicular occludin along with a loss of immunoreactive occludin in the seminiferous epithelium at the site of the BTB. Also, this CdCl(2)-induced occludin loss from the BTB coincided with a surge in testicular TGF-beta3, as well as p-p38 MAP kinase (the phosphorylated/activated form of p38), but not p38 MAP kinase and neither extracellular signal-regulated kinase nor its phosphorylated form (ERK/p-ERK), consistent with results of in vitro studies. More important, intratesticular administration of SB202190, a specific p38 MAP kinase inhibitor, could block the CdCl(2)-induced occludin loss from the BTB. These results illustrate that BTB dynamics in vivo are regulated by the TGF-beta3/p38 MAP kinase pathway, which in turn determines the level of occludin at the site of Sertoli cells TJs. PMID- 12639894 TI - Differential patterns of Fos induction in the hypothalamus of the rat following central injections of galanin-like peptide and galanin. AB - Galanin and its newly discovered relative galanin-like peptide (GALP) are neuropeptides that are implicated in the neuroendocrine regulation of body weight and reproduction. GALP has been shown to bind in vitro to galanin receptor subtypes 1 and 2, but whether it has its own specific receptor(s) is unknown. We reasoned that if GALP acts through a receptor that is distinct from galanin receptors, then GALP should activate central pathways that are different from those activated by galanin. The purpose of this study was to determine whether galanin and GALP produce different patterns of neuronal activation within the hypothalamus. Quantitative analysis of Fos immunoreactivity showed that galanin induced a significantly greater number of Fos-positive nuclei in the paraventricular nucleus compared with GALP (P < 0.001); however, compared with galanin, GALP induced significantly more Fos-positive cells in the horizontal limb of the diagonal band of Broca, caudal preoptic area, arcuate nucleus, and median eminence (P < 0.05). These observations suggest that GALP and galanin act through different receptor-mediated pathways to exert their effects on the regulation of body weight and reproduction and identify target cells for GALP's specific actions in the hypothalamus, including the preoptic area, paraventricular and arcuate nuclei, and the median eminence. PMID- 12639895 TI - Proteomic analysis of androgen-regulated protein expression in a mouse fetal vas deferens cell line. AB - During sex differentiation, androgens are essential for development of the male genital tract. The Wolffian duct is an androgen-sensitive target tissue that develops into the epididymis, vas deferens, and seminal vesicle. The present study aimed to identify androgen-regulated proteins that are involved in development of Wolffian duct-derived structures. We have used male mouse embryos transgenic for temperature-sensitive simian virus 40 large tumor antigen at 18 d of gestation, to generate immortalized mouse fetal vas deferens (MFVD) parental and clonal cell lines. The MFVD parental and clonal cell lines express androgen receptor protein and show features of Wolffian duct mesenchymal cells. Clonal cell line MFVD A6 was selected for proteomic analysis and cultured in the absence or presence of androgens. Subsequently, two-dimensional gel electrophoresis was performed on total cell lysates. Differentially expressed proteins were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and two androgen-regulated proteins were identified as mElfin and CArG-binding factor-A (CBF-A). CBF-A and mElfin are known to bind to cytoskeletal F-actin. Both proteins appeared to be regulated by androgens at the posttranslational level, possibly involving phosphorylation. Posttranslational modification of mElfin and CBF-A by androgens may be associated with a cytoskeletal change that is involved in androgen-regulated gene expression. PMID- 12639896 TI - Differential mechanisms and development of leptin resistance in A/J versus C57BL/6J mice during diet-induced obesity. AB - Changes in the biological efficacy of leptin were evaluated in obesity-resistant (A/J) and obesity-prone (C57BL/6J) mice at weaning and after consuming a high-fat (HF) diet for 4 and 8 wk. There was no evidence of leptin resistance in either strain at the start of the study, but after 4 and 8 wk on the HF diet, C57BL/6J mice became unresponsive to ip leptin. C57BL/6J mice responded to intracerebroventricular leptin at these time points but developed peripheral resistance to sympathetic stimulation of retroperitoneal white adipose tissue. In contrast, intracerebroventricular leptin was fully effective in A/J mice, reproducing the complete profile of responses observed in weanling mice. A/J mice were also partially responsive to ip leptin at both time points, increasing uncoupling protein 1 mRNA expression in brown adipose tissue and decreasing leptin mRNA in white adipose tissue. The findings indicate that retention of leptin responsiveness is an important component of the ability of A/J mice to mount a robust adaptive thermogenic response and resist diet-induced obesity. PMID- 12639897 TI - Dexamethasone induces rapid serine-phosphorylation and membrane translocation of annexin 1 in a human folliculostellate cell line via a novel nongenomic mechanism involving the glucocorticoid receptor, protein kinase C, phosphatidylinositol 3 kinase, and mitogen-activated protein kinase. AB - Our recent studies on rat pituitary tissue suggest that the annexin 1 (ANXA1) dependent inhibitory actions of glucocorticoids on ACTH secretion are effected via a paracrine mechanism that involves protein kinase C (PKC)-dependent translocation of a serine-phosphorylated species of ANXA1 (Ser-P-ANXA1) to the plasma membrane of the nonsecretory folliculostellate cells. In the present study, we have used a human folliculostellate cell line (PDFS) to explore the signaling mechanisms that cause the translocation of Ser-P-ANXA1 to the membrane together with Western blot analysis and flow cytometry to detect the phosphorylated protein. Exposure of PDFS cells to dexamethasone caused time dependent increases in the expression of ANXA1 mRNA and protein, which were first detected within 2 h of steroid contact. This genomic response was preceded by the appearance within 30 min of substantially increased amounts of Ser-P-ANXA1 and by translocation of the phosphorylated protein to the cell surface. The prompt membrane translocation of Ser-P-ANXA1 provoked by dexamethasone was inhibited by the glucocorticoid receptor, antagonist, mifepristone, but not by actinomycin D or cycloheximide, which effectively inhibit mRNA and protein synthesis respectively in our preparation. It was also inhibited by a nonselective PKC inhibitor (PKC(9-31)), by a selective inhibitor of Ca(2+)-dependent PKCs (Go 6976) and by annexin 5 (which sequesters PKC in other systems). In addition, blockade of phosphatidylinositiol 3-kinase (wortmannin) or MAPK pathways with PD 98059 or UO 126 (selective for MAPK kinse 1 and 2) prevented the steroid-induced translocation of Ser-P-ANXA1 to the cell surface. These results suggest that glucocorticoids induce rapid serine phosphorylation and membrane translocation of ANXA1 via a novel nongenomic, glucocorticoid receptor-dependent mechanism that requires MAPK, phosphatidylinositiol 3-kinase, and Ca(2+)-dependent PKC pathways. PMID- 12639898 TI - Inhibitory and stimulatory regulation of testicular inhibin B secretion by luteinizing hormone and follicle-stimulating hormone, respectively, in the rhesus monkey (Macaca mulatta). AB - This study examined the relative role of FSH and LH in governing testicular inhibin B secretion in the rhesus monkey. Adult male monkeys, rendered hypogonadotropic and hypogonadal by administration of a GnRH receptor antagonist (acyline), were implanted with testosterone (T)-filled or empty capsules. Following T-induced restoration of spermatogenesis, both groups received recombinant human FSH and vehicle for 12 d. Juvenile male monkeys received an 11 d infusion of single-chain recombinant human LH and recombinant human FSH, either alone or in combination. In adults, chronic hypogonadotropism resulted in a modest reduction of circulating inhibin B levels, which was more than fully reversed by FSH. In the presence of T, which exerted a marked suppression in inhibin B secretion, FSH restored inhibin B levels only to those observed before acyline treatment. In juveniles, treatment with single-chain recombinant human LH led to a suppression of inhibin B secretion and curtailed the FSH-induced stimulation of this testicular hormone. The T-induced decrease in inhibin B secretion was associated with suppression in inhibin-beta(B) mRNA levels, but FSH stimulation of inhibin B secretion occurred in the absence of clear changes in expression of this subunit gene. These findings indicate that inhibin B secretion by the monkey testis is governed by the inhibitory and stimulatory action of LH and FSH, respectively. The action of LH is presumably indirect and likely mediated by T inhibition of inhibin-beta(B) gene expression. The molecular basis of the stimulatory action of FSH on inhibin B secretion requires further study. PMID- 12639899 TI - Conservation of the hydroxysteroid sulfotransferase SULT2B1 gene structure in the mouse: pre- and postnatal expression, kinetic analysis of isoforms, and comparison with prototypical SULT2A1. AB - A novel mouse hydroxysteroid sulfotransferase cDNA has been cloned, and organization of its gene structure has been determined. The new mouse sulfotransferase, SULT2B1a, and its closely related isoform, SULT2B1b, are derived from a single SULT2B1 gene as a result of an alternative exon I and differential splicing. Thus, the only structural distinction between the two SULT2B1 isoforms is at their amino-terminal ends. Importantly, in contrast to the prototypical mouse hydroxysteroid sulfotransferase SULT2A1, the SULT2B1 isoforms have a predilection for cholesterol. Real-time RT-PCR reveals that the SULT2B1a isoform is most abundantly expressed in the brain and spinal cord, whereas SULT2B1b and SULT2A1 are weakly, if at all, expressed in the central nervous system. On the other hand, the SULT2B1b isoform is the most prominent hydroxysteroid sulfotransferase expressed in skin, whereas SULT2A1 is strikingly expressed in the liver. The substrate specificities and differential expression patterns of the three SULT2 isozymes strongly suggest that they have distinct biologic roles to play. Of further interest, the mouse SULT2B1 and SULT2A1 genes are differentially expressed during embryonic development, with the former being expressed at all stages from E8.5-E19, whereas the latter is not expressed until E19. It is speculated that, during embryonic development, SULT2B1b is required for production of cholesterol sulfate essential for normal skin development, whereas SULT2B1a produces pregnenolone sulfate, an essential neurosteroid during development of the central nervous system. PMID- 12639900 TI - Interleukin-18 is a novel mitogen of osteogenic and chondrogenic cells. AB - IL-18 was identified due to its ability to induce interferon-gamma (IFNgamma) production by T cells. It is a pleiotropic factor that shares structural features with IL-1 and functional activities with IL-12. IL-18 has a role in T cell development, where it has been demonstrated to act cooperatively with IL-12 to regulate IFNgamma. In bone, IL-18 is mainly produced by macrophages, but is also expressed by osteoblasts and inhibits osteoclast formation through granulocyte macrophage colony-stimulating factor (GM-CSF) and not IFNgamma production by T cells. We have investigated the effects of IL-18 on mature osteoclast activity and for potential actions on osteoblasts or chondrocytes. The effects of IL-18 on mature osteoclast activity were determined using two assays: isolated mature osteoclast cell culture and neonatal murine calvarial organ culture. IL-18 did not affect bone resorption in either assay system. The actions of IL-18 on osteogenic cells (primary cell cultures of fetal rat and neonatal mouse osteoblasts, as well as neonatal mouse calvarial organ culture) and primary chondrocytes (canine) were assessed by proliferation assays (quantification of cell numbers and thymidine incorporation). In each assay system, IL-18 acted as a mitogen to the osteogenic and chondrogenic cells. Since IL-18 signal transduction may involve IFNgamma or GM-CSF, we assessed their involvement in the IL-18 response. IL-18 did not induce IFNgamma production by primary osteoblasts, but, of greater significance, IFNgamma had the opposing action to IL-18 in that it inhibited the primary osteoblast cell proliferation. Although IL-18 rapidly induced GM-CSF production by primary osteoblasts, IL-18 was still mitogenic in osteoblast preparations established from GM-CSF-deficient mice. Combined, these studies indicate that IL-18 may have an autocrine/paracrine mitogen role for both osteogenic and chondrogenic cells, independent of the production of IFNgamma or GM-CSF. PMID- 12639901 TI - Novel neurotrophin-1/B cell-stimulating factor-3 (cardiotrophin-like cytokine) stimulates corticotroph function via a signal transducer and activator of transcription-dependent mechanism negatively regulated by suppressor of cytokine signaling-3. AB - Novel neurotrophin-1/B cell-stimulating factor-3 (NNT-1/BSF-3) is a recently cloned gp130 cytokine, acting through the tripartite ciliary neurotrophic factor receptor (CNTFR) alpha/leukemia inhibitory factor receptor (LIFR)/gp130 receptor complex. The aim of the current study was to investigate the role of NNT-1/BSF-3 in corticotroph cell function and further characterize NNT-1/BSF-3 signaling pathways. Using RT-PCR, expression of ciliary neurotrophic factor receptor alpha, leukemia inhibitory factor receptor, and gp130 could be demonstrated in mRNA derived from murine corticotroph AtT-20 cells and murine pituitary tissue. Incubation of AtT-20 cells with 10 ng/ml recombinant human NNT-1/BSF-3 rapidly induced tyrosine-phosphorylation of signal transducer and activator of transcription (STAT)3 and STAT1 at 5 and 10 min. Proopiomelanocortin promoter activity and suppressor of cytokine signaling (SOCS)-3 promoter activity were significantly stimulated by NNT-1/BSF-3 4.0 +/- 0.3- and 5.9 +/- 0.2-fold, respectively. In comparison with untreated control, NNT-1/BSF-3 significantly stimulated ACTH secretion at 24 and 48 h 1.7 +/- 0.2-fold and 1.5 +/- 0.1-fold above baseline. In comparison with mock-transfected cells, stable overexpression of SOCS-3 in AtT-20 cells abolished NNT-1/BSF-3-induced STAT1 and STAT3 phosphorylation and almost completely inhibited STAT-dependent proopiomelanocortin promoter and SOCS-3 promoter activities. In addition, NNT 1/BSF-3-induced ACTH secretion at 48 h was significantly attenuated by SOCS-3 overexpression. In summary, we have shown that NNT-1/BSF-3 is a modulator of corticotroph cell function, which is negatively regulated by SOCS-3. Our data indicate that the activation of the Jak-STAT cascade is essential for corticotroph NNT-1/BSF-3 signaling. Further studies will have to investigate the possible in vivo role of NNT-1/BSF-3 as a neuroimmunoendocrine modulator of hypothalamus-pituitary-adrenal axis stress response. PMID- 12639902 TI - Insulin receptor substrate-4 is expressed in muscle tissue without acting as a substrate for the insulin receptor. AB - Insulin receptor substrate (IRS) proteins represent key elements of the insulin signaling cascade. IRS-4 is the most recently characterized member of the IRS family with an undefined in vivo function. In contrast to IRS-1 and IRS-2, IRS-4 exhibits a limited tissue expression, and IRS-4 protein has not been detected in any mouse or primary human tissue so far. The purpose of the present study was to analyze the expression of IRS-4 in rat muscle and human skeletal muscle cells and assess involvement of IRS-4 in initial insulin signaling. Using immunoblotting and immunoprecipitation, the specific expression of IRS-4 protein could be demonstrated in rat soleus and cardiac muscle and human skeletal muscle cells, but it was not significantly detectable in quadriceps and gastrocnemius. A prominent down-regulation of IRS-4 was observed in heart and soleus muscle of WOKW rats, an animal model of the metabolic syndrome. In human skeletal muscle cells, both IRS-1 and IRS-2 are rapidly phosphorylated on tyrosine in response to insulin, whereas essentially no tyrosine phosphorylation of IRS-4 was observed in response to both insulin and IGF-I. Instead, a 2-fold increase in IRS-4 tyrosine phosphorylation was observed in myocytes subjected to osmotic stress. In conclusion, IRS-4 protein is expressed in heart and skeletal muscle in a fiber type specific fashion. Our data suggest that IRS-4 does not function as a substrate of the insulin and the IGF-I receptor in primary muscle cells but may be involved in nonreceptor tyrosine kinase signaling. PMID- 12639903 TI - Gonadal steroids differentially regulate the messenger ribonucleic acid expression of pituitary orexin type 1 receptors and adrenal orexin type 2 receptors. AB - Hypothalamic prepro-orexin as well as pituitary and adrenal orexin receptors are gender-specifically expressed. To assess the regulation by gonadal steroids, we investigated the effect of 17beta-estradiol in female and of testosterone in male rats on prepro-orexin and orexin receptor mRNA expression. Rats were either sham operated or gonadectomized and subsequently treated with placebo, 17beta estradiol, or testosterone for 21 d. Tissue mRNA levels of prepro-orexin, orexin type-1 (OX(1)), and orexin type-2 (OX(2)) receptors were measured using quantitative real-time RT-PCR. In female rats, pituitary OX(1) receptor mRNA levels were increased 12-fold after ovariectomy compared with sham- operated rats. The increase of pituitary OX(1) receptor mRNA was inhibited by treatment with 17beta-estradiol. Adrenal mRNA levels of OX(2) receptors in ovariectomized rats were increased 2-fold compared with sham-operated rats and were also reduced by treatment with 17beta-estradiol. In male rats, orchidectomy increased the mRNA levels of pituitary OX(1) receptors compared with sham-operated rats. In contrast, adrenal OX(2) receptor mRNA was reduced after orchidectomy. Testosterone treatment reversed the effect of orchidectomy on pituitary OX(1) and adrenal OX(2) receptors. In the hypothalamus, no differences were found in the mRNA levels of prepro-orexin, OX(1), and OX(2) receptors between sham-operated, placebo-treated, and steroid-treated female or male rats. Our results indicate that gonadal steroids differentially regulate pituitary OX(1) receptors and adrenal OX(2) receptors in male and female rats and may contribute to specific sex- dependent neuroendocrine and endocrine actions of orexins. PMID- 12639904 TI - Parathyroid hormone enhances mechanically induced bone formation, possibly involving L-type voltage-sensitive calcium channels. AB - PTH and mechanical loading might act synergistically on bone formation. We tested the in vivo effect of the L-type voltage-sensitive calcium channel (VSCC) blocker, verapamil, on bone formation induced by human PTH-(1-34) (PTH) injection with or without mechanical loading. Adult rats were divided into eight groups: vehicle, verapamil, PTH, or verapamil plus PTH with or without mechanical loading. Verapamil (100 mg/kg) was given orally 90 min before loading. PTH (80 micro g/kg) was injected sc 30 min before loading. Loading applied to tibia and ulna for 3 min significantly increased the bone formation rate on both the endocortical surface of tibia and the periosteal surface of ulna (P < 0.0001). Treatment with PTH enhanced load-induced bone formation by 53% and 76% (P < 0.001) on the endocortical and periosteal surfaces, respectively. Treatment with verapamil suppressed load-induced bone formation rate by 77% and 59% (P < 0.01). Furthermore, verapamil suppressed bone formation in rats subjected to PTH plus loading by 74% and 68% (P < 0.0001) at the tibia and ulna, respectively. In the groups without loading, neither verapamil nor PTH treatment significantly changed any bone formation parameter. This study indicates that L-type VSCCs mediate load induced bone formation in vivo. Furthermore, PTH enhances load-induced bone adaptation through involvement of L-type VSCCs. PMID- 12639905 TI - Estrogen signaling at the cell surface coupled to nitric oxide release in Mytilus edulis nervous system. AB - In previous studies we have demonstrated release of nitric oxide (NO) in human tissues following exposure to estrogen. We now designed experiments to determine whether estrogen is present in the neural tissue of Mytilus edulis, a marine mollusk, and whether, as in vertebrates, it stimulates constitutive NO synthase activity. After HPLC purification of 17beta-estradiol (17beta-E(2)) from M. edulis ganglionic tissue, we confirmed the presence of 17beta-E(2) by RIA and ES Q-TOF-MS analysis. We further found that when either exogenous or endogenous (purified HPLC fraction) 17beta-E(2) was added to pedal ganglia, there was immediate concentration-dependent NO release. Furthermore, 17beta-E(2) conjugated to BSA also stimulated NO release, suggesting mediation by a membrane surface receptor. Tamoxifen, an estrogen receptor antagonist, inhibited the action of both 17beta-E(2) and 17beta-E(2) conjugated to BSA, further supporting the presence of an estrogen receptor. In addition, by Western blot analysis with anti ER-beta antibodies, we observed a 55-kDa protein in both the membrane and cytosolic fractions in pedal ganglia as well as in human leukocytes (that have been previously shown to express ER-beta). In summary, our results suggest that a physiological dose of estrogen acutely stimulates NO release within pedal ganglia via an estrogen cell surface receptor. PMID- 12639906 TI - Effect of iodide on nicotinamide adenine dinucleotide phosphate oxidase activity and Duox2 protein expression in isolated porcine thyroid follicles. AB - Thyroperoxidase requires H(2)O(2) to catalyze the biosynthesis of thyroxine residues on thyroglobulin. Iodide inhibits the generation of H(2)O(2), and consequently the synthesis of thyroid hormones (Wolff-Chaikoff effect). The H(2)O(2) generator is a calcium-dependent nicotinamide adenine dinucleotide phosphate (NADPH) oxidase involving the flavoprotein Duox2. NADPH oxidase activity and Duox2 require cAMP to be expressed in pig thyrocytes. We studied the effect of iodide on NADPH oxidase activity, the DUOX2 gene, and Duox2 protein expression in pig thyroid follicles cultured for 48 h with forskolin or a cAMP analog. Iodide inhibited the cellular release of H(2)O(2) and NADPH oxidase activity, effects prevented by methimazole. Northern blot studies showed that iodide did not reduce DUOX2 mRNA levels but did reduce those of TPO and NIS. Western blot analyses using a Duox2-specific antipeptide showed that Duox2 has two N-glycosylation states, which have oligosaccharide motifs accounting for about 15 kDa and 25 kDa, respectively, of the apparent molecular mass. Cyclic AMP increased the amount of the highly glycosylated form of Duox2, an effect antagonized by iodide in a methimazole-dependent manner. These data suggest that an oxidized form of iodide inhibits the H(2)O(2) generator at a posttranscriptional level by reducing the availability of the mature Duox2 protein. PMID- 12639907 TI - Dynamics of Myc/Max/Mad expression during luteinization of primate granulosa cells in vitro: association with periovulatory proliferation. AB - Granulosa cell luteinization involves the attenuation of gonadotropin-induced proliferation. Although recent evidence indicates that primate granulosa cells stop dividing within 12 h of an ovulatory stimulus, early events in cell cycle arrest remain unknown. In the current study an in vitro model of primate granulosa cell luteinization is established that allows assessment of early events in terminal differentiation. A luteinizing dose of human chorionic gonadotropin (hCG) results in a secondary rise in proliferation before cell cycle arrest that is paralleled by a transient increase in the expression of c-Myc. In contrast, the c-Myc antagonists Mad1, Mad4, and Mxi1 are transiently repressed by hCG. Max, the common dimerization partner for Myc and Mad, is similarly repressed by hCG, suggesting that changes in the expression of this gene may further regulate the activity of Myc and Mad. To determine whether other cell cycle regulatory families are involved in luteinization, the expression of p53 and the wild-type p53-inducible phosphatase (wip1) was examined. Similar to Mad and Max, p53 and wip1 are transiently repressed by hCG, suggesting that the p53 and Mad pathways have either parallel or cooperative roles in luteinization. Thus, luteinization of primate granulosa cells is preceded by a burst of proliferation that is regulated by changes in the relative levels of c-Myc, Max, and Mad as well as p53. PMID- 12639908 TI - Identification of androgen-responsive genes in the rat ventral prostate by complementary deoxyribonucleic acid subtraction and microarray. AB - We have reported the identification of approximately two dozen androgen responsive genes, on the basis of their induction by androgens in the castrated rat ventral prostate, using PCR-based subtractive hybridization. The same prostatic cDNA samples were subjected to a modified subtractive hybridization, resulting in the identification of 21 new androgen-responsive genes, of which 14 were known and 7 were novel genes. To complement our subtraction study, we have used an Incyte rat cDNA microarray, consisting of 8951 known genes and expressed sequence tags, and found that 162 genes were up-regulated and 143 genes were down regulated 2.3-fold or more by androgens. As expected, the genes isolated from our subtraction overlap with genes found with microarray. Northern blot was carried out on all of the genes identified by subtraction and a few selected genes from microarray. All of the assayed genes are regulated by androgens, validating our subtraction and microarray studies. The identified genes can be classified into several functional groups, including metabolism, protein chaperoning and trafficking, protein synthesis, secretions, cell cycle and apoptosis, structural and extracellular matrix proteins, and novel proteins. Identification of these androgen-responsive genes will contribute to the understanding of androgen action in the normal and cancerous prostate. PMID- 12639909 TI - Nongenomic effect of aldosterone on Na+,K+-adenosine triphosphatase in arterial vessels. AB - Aldosterone increases Na(+),K(+)-adenosine triphophatase (Na(+),K(+)-ATPase) pump activity and abundance under chronic conditions in several tissues, including rat arterial vessels. The present study was undertaken to evaluate whether aldosterone has also short-term effects on the Na(+),K(+)-ATPase of rat aorta. The pump function was measured as ouabain-sensitive (86)Rb/K uptake in aortic rings. Addition of aldosterone induced a rapid inhibition of the Na(+),K(+) ATPase (57.0 +/- 2.3% of control values; P < 0.05; n = 8), followed by a return to control values after 120 min. The aldosterone-induced decrease in ouabain sensitive (86)Rb/K uptake was prevented by the new mineralocorticoid receptor antagonist eplerenone. The inhibition of gene transcription (actinomycin D) or protein synthesis (cycloheximide) had no effect on short-term aldosterone action on Na(+),K(+)-ATPase. The rapid aldosterone inhibition was also observed in the presence of monensin, a sodium-specific ionophore. Rapamycin, an immunosuppressive drug that stabilizes the heat shock protein-steroid receptor complex, blocked the rapid aldosterone effect. Bisindole I, an inhibitor of protein kinase C, also blocked nongenomic action of aldosterone on the Na pump. The nongenomic effect of aldosterone was inhibited by disrupters of microtubule (colchicine). Plasma membrane protein biotinylation of aortic segments and Western blot indicated a diminished presence of catalytic isoforms of Na(+),K(+) ATPase on the cell surface. Our findings indicate that aldosterone has a nongenomic effect on the Na(+),K(+)-ATPase of vascular tissue. This effect is mediated through protein kinase C activation and implies reduced cell surface abundance of catalytic subunits. These observations together with our previous report on chronic hormone replacement suggest that aldosterone is directly involved in ionic cellular homeostasis of the vascular system through Na pump regulation. PMID- 12639910 TI - Translational control of protein synthesis in muscle and liver of growth hormone treated pigs. AB - GH treatment increases protein deposition and the efficiency of dietary protein used for growth. To identify the mechanisms that regulate tissue protein synthesis in response to exogenous GH treatment, fully fed, growing swine were treated with GH for 7 d. Fasted and fed pigs were infused with [1-(13)C]leucine to determine protein synthesis rates, and translation initiation factor activity levels were measured in skeletal muscle and liver. Feeding increased protein synthesis and translational efficiency in both muscle and liver of control and GH treated pigs, and this was associated with increased 4E-BP1 and S6 kinase 1 phosphorylation, decreased association of eukaryotic initiation factor (eIF) 4E with 4E-BP1, and increased association of eIF4E with eIF4G. GH increased muscle protein synthesis and translational efficiency in fed pigs. GH increased liver protein synthesis of fasted and fed pigs in association with increased ribosome number. In muscle, but not liver, GH increased eIF2B activity and 4E-BP1 phosphorylation in both the fasted and fed state and increased the association of eIF4E with eIF4G in the fed state. We conclude that GH increases muscle protein synthesis in the fed state, in part, via mechanisms that enhance the binding of mRNA and methionyl-tRNA to the 40S ribosomal subunit, whereas GH increases liver protein synthesis in the fasted and fed states by increasing ribosome number. The results further indicate that the GH-induced protein synthetic response is dependent upon nutritional state and is tissue specific. PMID- 12639911 TI - Prostaglandin (PG) FP and EP1 receptors mediate PGF2alpha and PGE2 regulation of interleukin-1beta expression in Leydig cell progenitors. AB - Prostaglandins (PG) mediate IL-1beta regulation of several interleukin mRNAs in progenitor Leydig cells. PGE(2) and PGF(2alpha) potently reverse indomethacin (INDO; a cyclooxygenase inhibitor) inhibition of IL-1beta autoinduction. IL-1beta increases PGE(2) and PGF(2alpha) production. To determine the PG receptors involved in this regulation, this study established by RT-PCR and Western analyses which specific receptors for PGE(2) (EP receptors) and PGF(2alpha) (FP receptors) are expressed in progenitors. Pharmacological characterization of receptors involved in PGE(2) and PGF(2alpha) regulation of IL-1beta mRNA levels was ascertained using real-time PCR analyses. FP, EP(1), EP(2), and EP(4) receptor mRNAs and proteins, and an EP(3) receptor subtype were detected. IL 1beta treatment (24-h) significantly decreased EP(1) receptor levels; INDO abrogated this down-regulation. FP, EP(2), and EP(4) receptor levels increased after IL-1beta and IL-1beta + INDO. A selective FP agonist, cloprostenol (0.1 micro M), and PGF(2alpha) (10 micro M) had similar effects on IL-1beta mRNA levels in progenitors treated with IL-1beta + INDO. None of the EP(2)/EP(4) agonists [butaprost, misoprostol, or 11-deoxy PGE(1) (10 micro M)] affected IL 1beta mRNA levels. In contrast, EP(1)/EP(3) agonists (17-phenyl trinor PGE(2) and sulprostone) increased IL-1beta mRNAs in a dose-dependent manner. EP(1) receptor subtype-selective antagonist, SC-51322, blocked IL-1beta-induced and [IL-1beta + INDO + 17-phenyl trinor PGE(2)]-induced increases in IL-1beta mRNAs. Taken together, our data demonstrate that FP and EP(1) receptors mediate PGF(2alpha) and PGE(2) induction of progenitor IL-1beta expression. PMID- 12639912 TI - Cyclic adenosine 3',5'-monophosphate regulation of corticotropin-releasing hormone promoter activity in AtT-20 cells and in a transformed hypothalamic cell line. AB - The regulation of CRH promoter activity by cAMP was studied in two cell lines, the pituitary corticotroph cell line AtT-20 and the immortalized hypothalamic cell line 4B, which expresses CRH and vasopressin. In 4B cells transfected with a CRH promoter-luciferase construct, the adenylyl cyclase stimulator, forskolin, increased luciferase activity in parallel with increases in intracellular cAMP. In 4B cells, however, the phosphodiesterase inhibitor, isobutylmethylxanthine, potentiated forskolin-stimulated cAMP without affecting further increases in luciferase activity. In AtT-20 cells, forskolin plus isobutylmethylxanthine elevated cAMP only slightly, but increased luciferase activity to levels similar to those observed in 4B cells. AtT-20 cells were also unresponsive to 8-bromo cAMP, due in part to higher phosphodiesterase (PDE) activities. Although both cells contained PDE1, -3, and -4, inhibition of either PDE4 or PDE1 potentiated luciferase activity stimulated by submaximal forskolin concentrations in 4B cells, while only simultaneous inhibition of PDE3 and PDE4 was effective in AtT 20 cells. The data show that minor elevations in intracellular cAMP are sufficient for full stimulation of CRH promoter activity regardless of the cell line. Furthermore, poor CRH promoter activation in AtT-20 cells appears to result from deficient cAMP production and rapid cAMP degradation by PDE. PMID- 12639913 TI - Regulation of melanocortin-4 receptor signaling: agonist-mediated desensitization and internalization. AB - Disruption of the hypothalamic melanocortin-4 receptor (MC4R) pathway results in obesity both in humans and rodents, demonstrating a crucial role for hypothalamic MC4Rs in the regulation of energy homeostasis. Because even haploinsufficiency of the MC4R gene can cause obesity in humans and mice, subtle changes in receptor numbers or signaling are likely to impact upon the regulation of food intake and energy expenditure. Little is known about the intracellular regulation of MC4R signaling. Using GT1-7 cells, we show for the first time that the MC4R undergoes ligand-mediated desensitization. We then addressed the possible mechanisms underlying the desensitization using HEK293 and COS-1 cells transfected with hemagglutinin-tagged human MC4R. Preexposure of GT1-7 cells that express endogenous MC4R to the agonist for MC4R, alpha-melanocyte-stimulating hormone, resulted in impaired cAMP formation to a second challenge of alpha-melanocyte stimulating hormone. The desensitization of MC4R was accompanied by time dependent internalization of the receptor in HEK293 cells, which was partly inhibited by pretreatment with a specific protein kinase A (PKA) inhibitor, H89. In COS-1 cells, overexpression of dominant-negative G protein-coupled receptor kinase (GRK) 2-K220R partly inhibited the agonist-mediated internalization of MC4R, whereas it did not in HEK293 cells. Overexpression of dominant-negative mutants of beta-arrestin1-V53D and dynamin I-K44A prevented agonist-mediated internalization of MC4R. Mutagenesis studies revealed that Thr312 and Ser329/330 in the C-terminal tail are potential sites for PKA and GRK phosphorylation and may play an essential role in the recruitment of beta-arrestin to the activated receptor. Our data demonstrate that, through PKA-, GRK-, beta-arrestin-, and dynamin-dependent processes, MC4R undergoes internalization in response to agonist, thereby providing novel insights into the regulation of MC4R signaling. PMID- 12639914 TI - Thyroid hormone transport by the rat fatty acid translocase. AB - We examined the hypothesis that rat fatty acid translocase (rFAT) mediates the cellular uptake of T(3) and other iodothyronines. Uninjected Xenopus laevis oocytes and oocytes injected 4 d previously with rFAT cRNA were incubated for 60 min at 25 C in medium containing 0.01-10 micro M [(125)I]T(3) and 0.1% BSA, or 1 100 micro M [(3)H]oleic acid and 0.5% BSA. Injection of rFAT cRNA resulted in a 1.9-fold increase in uptake of T(3) (10 nM) and a 1.4-fold increase in uptake of oleic acid (100 micro M). Total T(3) uptake was lower in the presence than in the absence of BSA, but relative to the free T(3) concentration, uptake was increased by BSA. The fold induction of T(3) uptake by rFAT was not influenced by BSA. By analyzing uptake as a function of the ligand concentration, we estimated a K(m) value of 3.6 micro M for (total) T(3) and 56 micro M for (total) oleic acid. In addition to T(3), rFAT mediates the uptake of T(4), rT(3), 3,3'-diiodothyronine, and T(3) sulfate. The injection of human type III deiodinase cRNA with or without rFAT cRNA resulted in the complete deiodination of T(3) taken up by the oocytes, indicating that T(3) is indeed transported to the cytoplasm. In conclusion, our results demonstrate transport of T(3) and other iodothyronines by rFAT. PMID- 12639915 TI - Targeted restoration of cleavage in a noncleaving thyrotropin receptor demonstrates that cleavage is insufficient to enhance ligand-independent activity. AB - Two unusual features of the TSH receptor (TSHR) ectodomain are its intramolecular cleavage at the cell surface into disulfide-linked subunits and its constraint of ligand-independent (constitutive) activity inherent to the serpentine region. Whether ectodomain cleavage alters the level of TSHR constitutive activity is an important unanswered question. To address this issue, we used a TSHR engineered so as not to undergo spontaneous cleavage into subunits (deletion of amino acid residues 317-366 and GQE(367-369)NET substitution). Into this noncleaving TSHR (termed TSHR-D1-NET), we introduced thrombin recognition motifs (termed Thr 6 and Thr 18) at the site of spontaneous cleavage. Treatment of intact Chinese hamster ovary cells expressing TSHR-D1-NET-Thr 6 and -Thr 18 with thrombin induced cleavage into A and B subunits, as determined by (125)I-TSH covalent cross linking. Nevertheless, constitutive activity of the thrombin-cleaved TSHR was unaltered. The level of TSHR constitutive activity was, therefore, fully dissociated from intramolecular cleavage into subunits. Trypsin treatment of the same cells expressing the noncleaving TSHR also generated disulfide-linked A and B subunits but, in contrast to thrombin, enhanced TSHR constitutive activity. Therefore, the activating effect of trypsin appears to involve clipping at an additional, as-yet unidentified, site. In summary, our data demonstrate that TSHR cleavage is, by itself, insufficient to reduce TSHR ectodomain constraint on ligand-independent constitutive activity. These data are consistent with other evidence that A subunit shedding consequent to TSHR cleavage is a critical factor in enhancing TSHR constitutive activity. PMID- 12639916 TI - Hypothalamic proopiomelanocortin neurons are glucose responsive and express K(ATP) channels. AB - Hypothalamic proopiomelanocortin (POMC) neurons are critical for controlling homeostatic functions in the mammal. We used a transgenic mouse model in which the POMC neurons were labeled with enhanced green fluorescent protein to perform visualized, whole-cell patch recordings from prepubertal female hypothalamic slices. The mouse POMC-enhanced green fluorescent protein neurons expressed the same endogenous conductances (a transient outward K(+) current and a hyperpolarization-activated, cation current) that have been described for guinea pig POMC neurons. In addition, the selective micro -opioid receptor agonist DAMGO induced an outward current (maximum of 12.8 +/- 1.2 pA), which reversed at K(+) equilibrium potential (E(K+)), in the majority (85%) of POMC neurons with an EC(50) of 102 nM. This response was blocked by the opioid receptor antagonist naloxone with an inhibition constant of 3.1 nM. In addition, the gamma aminobutyric acid(B) receptor agonist baclofen (40 micro M) caused an outward current (21.6 +/- 4.0 pA) that reversed at E(K+) in these same neurons. The ATP sensitive potassium channel opener diazoxide also induced an outward K(+) current (maximum of 18.7 +/- 2.2 pA) in the majority (92%) of POMC neurons with an EC(50) of 61 micro M. The response to diazoxide was blocked by the sulfonylurea tolbutamide, indicating that the POMC neurons express both Kir6.2 and sulfonylurea receptor 1 channel subunits, which was verified using single cell RT PCR. This pharmacological and molecular profile suggested that POMC neurons might be sensitive to metabolic inhibition, and indeed, we found that their firing rate varied with changes in glucose concentrations. Therefore, it appears that POMC neurons may function as an integrator of metabolic cues and synaptic input for controlling homeostasis in the mammal. PMID- 12639917 TI - Role of the different mitogen-activated protein kinase subfamilies in the stimulation of dog and human thyroid epithelial cell proliferation by cyclic adenosine 5'-monophosphate and growth factors. AB - We have investigated the role of the different classes of MAPKs, i.e. ERKs, c-Jun N-terminal kinases (JNKs), and p38 MAPK in the proliferation of dog and human thyroid epithelial cells (thyrocytes) in primary cultures. In these cells, TSH, acting through cAMP, epidermal growth factor, hepatocyte growth factor (HGF), and phorbol 12-myristate 13-acetate induce DNA synthesis. With the exception of HGF, all of these factors require the presence of insulin for mitogenic effects to be expressed. We found that TSH and forskolin are without effect on the phosphorylation and activity of the different classes of MAPKs. In contrast, all the cAMP-independent growth factors, whereas without effect on the phosphorylation and activity of JNKs and p38 MAPK, stimulated the ERKs. This effect was strong and sustained in response to HGF, epidermal growth factor and 12-myristate 13-acetate but weak and transient in response to insulin. Moreover, whereas in stimulated cells DNA synthesis was inhibited by PD 098059, an inhibitor of MAPK kinase 1 and consequently of ERKs, it was not modified by SB 203580, an inhibitor of p38 MAPK. Taken together, these data 1) exclude a role of JNKs and p38 MAPK in the proliferation of dog and human thyrocytes; 2) suggest that the mitogenic action of the cAMP-independent agents requires a strong and sustained activation of both ERKs and phosphatidylinositol 3-kinase/protein kinase B as realized by HGF alone or by the other agents together with insulin; and 3) show that TSH and cAMP do not activate ERKs but that the weak activation of ERKs by insulin is nevertheless necessary for DNA synthesis to occur. PMID- 12639918 TI - Estrogen regulates adrenal angiotensin type 1 receptors by modulating adrenal angiotensin levels. AB - Estrogen inhibits adrenal angiotensin type 1 receptor (AT(1)R) binding sites and attenuates the adrenal responsivity to angiotensin II (Ang II). Ang II modulates AT(1)R expression. Here, we determined if estrogen-induced down-regulation of adrenal AT(1)Rs involves modulation of adrenal Ang II. Female rats were ovariectomized (OVX) and injected with 17beta-estradiol benzoate (E(2); 40 micro g/kg) or vehicle for 7 d. Adrenal Ang II was separated from other angiotensin peptides by HPLC and measured by RIA. Scatchard analysis of radioligand binding curves showed that E(2) or captopril (Cap; 0.5 g/liter water) significantly reduced adrenal AT(1)R binding (maximum binding capacity) by 22% and 19%, respectively, compared with OVX (276 +/- 2.09 fmol/mg protein). E(2) and Cap lowered adrenal Ang II levels by 39% and 21%, respectively, compared with OVX (4.10 +/- 0.44 pmol/g). E(2) caused no further reductions in adrenal AT(1)R binding or in Ang II levels in Cap-treated OVX rats. High-dose Ang II infusion (1000 ng/kg.min) increased adrenal Ang II levels by 71% and lowered AT(1)R binding by 18%. Under these infusion conditions, E(2) did not reduce adrenal Ang II or AT(1)R binding. No differences in AT(1)R affinity (dissociation constant) were observed among groups. These data suggest that E(2) regulates the number of adrenal AT(1)R binding sites indirectly by modulating adrenal Ang II. PMID- 12639920 TI - Differential gene expression in well-regulated and dysregulated pancreatic beta cell (MIN6) sublines. AB - To identify genes involved in regulated insulin secretion, we have established and characterized two sublines derived from the mouse pancreatic beta-cell line MIN6, designated B1 and C3. They have a similar insulin content, but differ in their secretory properties. B1 responded to glucose in a concentration- and cell confluence-dependent manner, whereas C3 did not. B1 cells were stimulated by phorbol 12-myristate 13-acetate, leucine, arginine, glibenclamide, isobutylmethylxanthine, and KCl, whereas C3 did not respond (leucine, arginine, and glibenclamide) or responded to a lesser extent (isobutylmethylxanthine, phorbol 12-myristate 13-acetate, and KCl). Although intracellular Ca(2+) rose in response to glucose in B1 but not C3 cells, KCl increased intracellular Ca(2+) in a similar manner in both sublines. GLUT-1, GLUT-2, Kir6.2, and SUR1 expression was not significantly different between B1 and C3 cells, whereas E-cadherin was more abundantly expressed in B1 cells. A more complete list of differentially expressed genes was established by suppression subtractive hybridization and high density (Affymetrix) oligonucleotide microarrays. Genes were clustered according to known or putative function. Those involved in metabolism, intracellular signaling, cytoarchitecture, and cell adhesion are of potential interest. These two sublines should be useful for identification of the genes and mechanisms involved in regulated insulin secretion of the pancreatic beta-cell. PMID- 12639919 TI - Immunotoxin lesion of hypothalamically projecting norepinephrine and epinephrine neurons differentially affects circadian and stressor-stimulated corticosterone secretion. AB - Hindbrain norepinephrine (NE) and epinephrine (E) neurons play a pivotal role in the central distribution of sensory signals derived from the internal environment. Their projections influence the various secretory patterns of the hypothalamo-pituitary-adrenal axis and are essential for feeding and adrenal medullary responses to glucoprivation. NE and E terminals in the paraventricular nucleus of the hypothalamus (PVH) and associated hindbrain cell bodies can be virtually eliminated by PVH microinjection of a retrogradely transported conjugate of saporin (SAP, a ribosomal toxin) and a monoclonal antibody against dopamine beta-hydroxylase (dbetah), i.e. dbetah mouse monoclonal antibody conjugated to SAP (DSAP). To examine the effects of selective elimination of NE/E afferents on hypothalamo-pituitary-adrenal activation, we injected DSAP into the PVH and measured corticosterone secretion under basal circadian conditions and in response to two distinct challenges: glucoprivation and forced swim. DSAP lesions profoundly impaired glucoprivation-induced corticosterone secretion and induction of CRH heteronuclear RNA and Fos mRNA in the PVH, without impairing basal CRH mRNA expression, circadian corticosterone release, or the corticosterone response to swim stress. Thus, NE/E projections influence corticosterone secretion only in certain circumstances. They are required for the response to glucoprivation, but are dispensable for circadian activation and for the response to swim stress. PMID- 12639922 TI - Gene therapy of a rat follicular thyroid carcinoma model with adenoviral vectors transducing murine interleukin-12. AB - Interleukin-12 (IL12) is a heterodimeric cytokine that plays an important role in the development of cellular immunity. We previously reported the antitumor activity of mouse IL12 (mIL12) transduced by adenovirus in a medullary thyroid carcinoma model. In this study, a rat thyroid follicular cancer cell line (RTC R2) was employed to develop tumors after sc injection in Wistar rats. In five of five animals, RTC-R2 cells infected with mIL12 transducing adenovirus (AdCMVmIL12) in vitro failed to be tumorigenic in vivo in syngenic rats, whereas four of five animals developed tumors after injection of luciferase transducing adenovirus (AdCMVLuc)-infected cells. After intratumoral treatment with AdCMVmIL12 at 1 x 10(9) plaque-forming units per rat, 90% (26/29) of animals bearing small (<100 mm(3)) tumors were apparently cured. Larger tumors treated by injection of AdCMVmIL12 became significantly smaller than AdCMVLuc-treated animals, and growth stabilized. Challenge studies showed that only 3 of 28 animals previously treated and cured with AdCMVmIL12 developed a tumor after sc reinjection of RTC-R2 cells, whereas all animals developed tumors in naive animals. Thus, AdCMVmIL12-treated animals developed long-term antitumor immunity. We also studied animals with two tumors, injecting virus in one. Tumors regressed at both sites in five of six animals after treatment of one tumor with AdCMVmIL12, and in the sixth animal one site tumor regressed and another tumor continued to grow. In AdCMVLuc-treated animals, both tumors regressed in only one animal, and the reminder continued to grow. To detect toxicity to liver and other tissues after administration of AdCMVmIL2, the vector was administrated intratumorally or iv at the dose of 2 x 10(9) plaque-forming units per rat. No change in behavior was observed in any of the treated animals. Rats were killed at different time after virus administration. An overt increase of spleen size was observed 7 d after infection in all animals treated with AdCMVmIL12. All animals given virus IV had some lymphocyte infiltration in the sinusoids and triads of the liver, whereas AdCMVmIL12 injected intratumorally did not cause this effect. Spleens of some virus-treated animals showed decreased white pulp, with apparently increased hematopoiesis. No specific changes were found in lungs and kidneys. Iv administration of AdCMVmIL12 induced a moderate increase of glutamic-oxalacetic transaminase and glutamic-pyruvic transaminase, whereas AdCMVmIL12 injected intratumorally did not. This study confirms the efficient antitumor activity of an adenovirus expressing mIL12 after in vivo delivery in an animal model and indicates the possibility of application to patients because of the low toxicity. PMID- 12639921 TI - Neuroendocrine control of follicle-stimulating hormone (FSH) secretion: III. Is there a gonadotropin-releasing hormone-independent component of episodic FSH secretion in ovariectomized and luteal phase ewes? AB - Our previous studies in ovariectomized ewes have provided direct evidence that FSH secretion is comprised of basal and episodic modes. In those studies, each GnRH pulse coincided with an FSH pulse, but additional FSH pulses were noted. To determine whether non-GnRH-associated pulses of FSH represent a GnRH-independent component of FSH secretion, we determined whether episodic FSH secretion persists after blockade of GnRH action with a GnRH antagonist. Hypophyseal portal and jugular blood was collected from five ovariectomized and six luteal phase ewes at 5-min intervals for 6 h before and 6 h after a single iv injection of Nal-Glu (10 micro g/kg body weight). Hypophyseal portal LH and FSH and jugular patterns of FSH were compared with patterns of GnRH. Before Nal-Glu, in both models, there was a one-to-one concordance between GnRH and portal LH pulses, and each GnRH pulse was associated with a FSH pulse. However, additional non-GnRH-associated pulses of FSH were present. Nal-Glu administration eliminated LH but not FSH pulsatility. Nal-Glu inhibited interaction of GnRH I with GnRH type I receptor but not interaction of GnRH II with type II receptor. These studies provide the first direct evidence of the existence of an acute GnRH I-independent component of episodic FSH secretion. PMID- 12639923 TI - Adrenocorticotropin induces mitogen-activated protein kinase phosphatase 1 in Y1 mouse adrenocortical tumor cells. AB - ACTH signaling pathway includes the action of both protein kinases, mainly cAMP dependent protein kinase (protein kinase A, PKA), and serine/threonine and tyrosine phosphatases. MAPK phosphatase-1 (MKP-1) is a dual activity protein phosphatase involved in the dephosphorylation of MAPK. To determine whether MKP-1 is a component of ACTH cascade, here we investigate the expression levels of MKP 1 gene in Y1 mouse adrenocortical tumor cells under ACTH stimulation. ACTH transiently increased MKP-1 mRNA and protein levels. MKP-1 mRNA increase occurred at 30 min, peaked at 1 h (6-fold), and returned to basal levels thereafter. The ACTH-mediated mRNA increase was blunted by actinomycin D and enhanced by cycloheximide. A cell permeable cAMP analog, 8-bromo-cAMP, also transiently induced MKP-1 mRNA (4-fold) and the PKA inhibitor N-[2-(p bromocinnamylamino)ethyl]-5-isoquinolinesulfonamid abolished this effect. In contrast, N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamid only partially reduced the effect of ACTH, suggesting the participation of PKA independent mechanisms in the hormone-induced MKP-1 expression. In addition, we show that the rise in intracellular Ca(2+) and protein kinase C activation had a potent synergic effect on ACTH- and 8-bromo-cAMP-mediated MKP-1 induction. In summary, our findings demonstrate that MKP-1 is another component of ACTH signaling cascade and indicate that this hormone may potentially down-regulate MAPKs. PMID- 12639924 TI - Mitogen-activated protein kinases potentiate thyroid hormone receptor transcriptional activity by stabilizing its protein. AB - Transcriptional regulation of downstream gene expression by thyroid hormone (T(3)) is mediated by the thyroid hormone receptor (TR). T(3) binding induces a complicated transition, where TR converts from a transcriptional repressor into a transcriptional activator and instigates downstream gene transcription. Binding of T(3) to TR also induces the degradation of TR, resulting in desensitization of the cells to further T(3) treatment. It has been shown that phosphorylation of TR plays a critical role in its activity and stability after T(3) binding. However, the kinases in control of phosphorylating TR in the nucleus have not been identified. In this study we demonstrate that MAPKs are possible candidates responsible for the nuclear phosphorylation of TR. Suppression of MAPKs with specific inhibitors repressed TR transcriptional activity and antagonized okadeic acid-induced TR transcriptional activity potentiation. Overexpression of the MAPK activator, MKK6, and its constitutively active mutant, MKK6EE, significantly increased TR activity and protected TR from degradation. Involvement of the 26S ubiquitin proteasome in hormone binding-induced TR degradation was also examined. We found that MAPKs enhanced the DNA binding affinity of TR. Our results suggest that MAPKs are the major kinases responsible for the nuclear phosphorylation of TR and are critical factors modulating the transcriptional activity and protein stability of TR subsequent to ligand binding. PMID- 12639926 TI - Prenatal programming of reproductive neuroendocrine function: fetal androgen exposure produces progressive disruption of reproductive cycles in sheep. AB - In the agonadal, androgenized ewe testosterone before birth produces a precocious pubertal rise in circulating LH and abolishes the LH surge mechanism. The present study tested two predictions from this model in the ovary-intact female: 1) prenatal androgen exposure produces early ovarian stimulation; and 2) despite early ovarian stimulation, progestogenic cycles would not occur because of the abolition or disruption of the LH surge. Pregnant ewes were injected with testosterone propionate twice per week from either d 30-90 (T60 group; 100 mg/injection) or d 60-90 (T30 group; 80 mg/injection) of gestation (term, 147 d). Control ewes received no injections. At birth, the androgenized and control lambs were divided into two groups: ovary-intact to determine the effects of prenatal androgen on the timing of puberty and subsequent ovarian function, and ovariectomized to assess the timing of the pubertal decrease in sensitivity to estrogen negative feedback and the subsequent increase in LH. Neonatally orchidectomized, estrogen-treated males were included for comparison of the timing of this pubertal rise in LH secretion. Neuroendocrine puberty (determined on the basis of LH increase) was advanced in the androgenized females to a similar age as in males. Repeated progesterone cycles of the same duration and number occurred in the ovary-intact ewes, and they began at the same time as for control females, thus negating both predictions. Differences appeared during the second breeding season, when reproductive cycles were either absent (T60) or disrupted (T30 group). Our findings reveal that exposure to androgens in utero causes a progressive loss of cyclic function in adulthood. PMID- 12639925 TI - Paraventricular nucleus administration of calcitonin gene-related peptide inhibits food intake and stimulates the hypothalamo-pituitary-adrenal axis. AB - Calcitonin gene-related protein (CGRP) inhibits food intake and stimulates the hypothalamo-pituitary-adrenal (HPA) axis after intracerebroventricular injection in rats. However, the hypothalamic site and mechanism of action are unknown. We investigated the effects of intraparaventricular nucleus administration (iPVN) of CGRP on food intake and the HPA axis in rats and the effect of CGRP on the release of hypothalamic neuropeptides in vitro. In addition, we investigated the effects of food deprivation on hypothalamic CGRP expression. CGRP dose dependently reduced food intake in the first hour after iPVN injection in fasted male rats (saline, 5.1 +/- 0.8 g; 0.3 nmol CGRP, 1.1 +/- 0.5 g; P < 0.001 vs. saline). iPVN injection of CGRP(8-37) (a CGRP(1) receptor antagonist) alone had no effect on food intake. However, the reduction in food intake by iPVN CGRP was attenuated by prior administration of CGRP(8-37) [CGRP(8-37) (10 nmol)/CGRP (0.3 nmol), 3.0 +/- 0.8 g; P < 0.05 vs. 0.3 nmol CGRP]. CGRP (100 nM) stimulated the release of alpha-melanocyte stimulating hormone, cocaine- and amphetamine-related transcript, corticotropin-releasing hormone, and arginine vasopressin from hypothalamic explants to 127 +/- 19%, 148 +/- 10%, 158 +/- 17%, and 198 +/- 21% of basal levels, respectively (P < 0.05 vs. basal), but did not alter the release of either neuropeptide Y or agouti-related protein. Hypothalamic CGRP mRNA levels in 24-h fasted rats were increased to 130 +/- 8% of control levels [CGRP mRNA (arbitrary units), 4.75 +/- 0.4; controls, 3.65 +/- 0.34; P < 0.05]. Our data suggest that CGRP administered to the PVN inhibits food intake and stimulates the HPA axis. PMID- 12639928 TI - Glucagon-like peptide-1 inhibits apoptosis of insulin-secreting cells via a cyclic 5'-adenosine monophosphate-dependent protein kinase A- and a phosphatidylinositol 3-kinase-dependent pathway. AB - The activation of the glucagon-like peptide-1 (GLP-1) receptor has been shown to have an important role in the functional activity of islet beta-cells and in the expansion of the islet cell mass. Constant remodeling of islet cell mass is mediated in vivo by proliferative and apoptotic stimuli to ensure a dynamic response to a changing demand for insulin. The present study was undertaken to investigate the biological activity of GLP-1 when cells were challenged by a proapoptotic stimulus. We have shown that activation of the GLP-1 receptor inhibits H(2)O(2)-induced apoptosis in a cultured mouse insulinoma cell line, termed MIN6. GLP-1 reduced DNA fragmentation and improved cell survival. This was mediated by an increased expression of the antiapoptotic proteins Bcl-2 and Bcl xL. GLP-1 also prevented the H(2)O(2)-dependent cleavage of poly-(ADP-ribose) polymerase. Inhibition of the GLP-1-dependent increase of cAMP by Rp-cAMP blocked the antiapoptotic action of GLP-1, as determined by DNA fragmentation and poly (ADP-ribose)-polymerase assays and by detection of Bcl-2 and Bcl-xL protein levels. Investigation of the role of the protein kinases, PI-3 kinase (PI3K) and MAPK, by use of the inhibitors PD098059 and LY294002 demonstrated that the activation of PI3K, but not MAPK, was required to prevent proapoptotic events in cells exposed to H(2)O(2). The present study provides evidence that GLP-1 has an antiapoptotic action mediated by a cAMP- and PI3K-dependent signaling pathway. PMID- 12639927 TI - Spatial distribution of the messenger ribonucleic acid and protein of the nuclear receptor coactivator, amplified in breast cancer-3, in mice. AB - Transcriptional activities of nuclear receptors are modulated by coactivators and corepressors. The amplified in breast cancer-3 protein (AIB3, also known as ASC 2, RAP250, PRIP, TRBP, and NCR) is a newly identified nuclear receptor coactivator that is amplified and overexpressed in breast cancers. This study aims to investigate the spatial expression of AIB3 mRNA and protein in various murine tissues. Quantitative measurements revealed that the concentrations of AIB3 mRNA differ substantially in different tissues in a descending order from the following: testis, brain, thymus, white fat, pituitary, ovary, adrenal gland, lung, uterus, kidney, heart, skeletal muscle, liver, and virgin mammary gland. The AIB3 mRNA level in the testis is 165-fold higher than that in the virgin mammary gland. Specific antiserum was generated and used to map the distribution of AIB3 protein by immunohistochemistry. Although AIB3 protein was detected in many tissues, the AIB3 immunoreactivities varied significantly from cell type to cell type. High levels of AIB3 immunoreactivity were observed in hormone target cells including the testicular Sertoli cells, follicular granulosa cells, and epithelial cells of the prostate, uterus, mammary gland, and kidney tubules. Medium and low levels of AIB3 immunoreactivities were also detected in a variety of other cell types. These results demonstrate that AIB3 mRNA and protein are preferentially expressed in specific cell types, suggesting that AIB3 may support the function of nuclear receptors in a cell type-specific manner. PMID- 12639929 TI - Cytotoxic activity of gonadotropin-releasing hormone (GnRH)-pokeweed antiviral protein conjugates in cell lines expressing GnRH receptors. AB - Pokeweed antiviral protein (PAP), a 29-kDa ribosome-inactivating protein isolated from the leaves of Phytolacca americana, has potent cytotoxic activity once it enters the cytoplasm of a cell. It is incapable of entering cells by itself. Therefore, our objective was to determine whether a GnRH analog could be used to deliver PAP specifically to cells expressing GnRH receptors. D-Lys(6)-GnRH-Pro(9) ethylamide was conjugated to PAP (GnRH-PAP). Chinese hamster ovary cells stably transfected with cDNA for the murine GnRH receptor and a mouse gonadotroph tumor cell line that expresses endogenous GnRH receptors (alphaT3-1 cells) were used to evaluate the cytotoxic effects of GnRH-PAP. We also examined cytotoxicity of GnRH PAP using human endometrial, breast, and prostate cancer cell lines. Treatment of GnRH receptor-positive cells with GnRH-PAP resulted in dose-dependent cytotoxicity. Cytotoxicity of GnRH-PAP was dependent on number of GnRH receptors (r(2) = 0.871, P < 0.05) and duration of exposure of GnRH-PAP to the cells. In contrast, GnRH-PAP was not cytotoxic to Chinese hamster ovary cells not harboring GnRH receptors. Moreover, the cytotoxic activity of GnRH-PAP could be inhibited by addition of excess GnRH analog. Neither PAP nor GnRH analog alone was cytotoxic. These results suggest that GnRH analogs can be used to specifically deliver toxin molecules to cells that express GnRH receptors. Thus, a new class of biomedicines that act as hormonotoxins against cells expressing GnRH receptors provides a novel approach for inhibiting reproduction and treating cancers that are dependent on reproductive hormones. PMID- 12639930 TI - Enhanced expression of uterine stathmin during the process of implantation and decidualization in rats. AB - We used the library subtraction technique to identify genes specifically expressed in the rat uterus during early pregnancy. One such gene was that for stathmin, a factor that is associated with tubulin binding and the destabilization of microtubules. Stathmin was expressed at higher levels in implantation sites than in interimplantation sites on d 6 and 7 of pregnancy; the levels on d 6 and 7 were higher in implantation sites than in the entire uterus on d 3-5 of pregnancy or in nonpregnant uteri. Intense expression of stathmin mRNA was primarily limited to the subluminal stromal cells at the implantation site. Expression was also detected in the decidual zones and was accentuated during the period of decidualization (d 7-12). In the delayed implantation pregnant rat model, uterine stathmin expression was low, but increased after implantation induced by administration of 17beta-estradiol to the progesterone primed animal. Further, decidualization in the pseudopregnant rat, induced by intrauterine infusion of oil, enhanced stathmin expression. Stathmin expression clearly increases in the uterus when stimulated by embryo implantation and decidualization and may play a role in the early stages of pregnancy. PMID- 12639931 TI - Cellular and molecular characterization of the adipose phenotype of the aromatase deficient mouse. AB - Estrogen deficiency in the aromatase knockout (ArKO) mouse leads to the development of obesity by as early as 3 months of age, which is characterized by a marked increase in the weights of gonadal and infrarenal fat pads. Humans with natural mutations of the aromatase gene also develop a metabolic syndrome. In the present study cellular and molecular parameters were investigated in gonadal adipose tissue from 10-wk-old wild-type (WT) and ArKO female mice treated with 17beta-estradiol or placebo to identify the basis for the increase in intraabdominal obesity. Stereological examination revealed that adipocytes isolated from ArKO mice were significantly larger and more abundant than adipocytes isolated from WT mice. Upon treatment with estrogen, the volume of these adipocytes was greatly reduced, whereas the reduction in the number of adipocytes was much less pronounced. Transcriptional analysis using real-time PCR revealed concomitant changes with adipocyte volume in the levels of transcripts encoding leptin and lipoprotein lipase, whereas peroxisome proliferator-activated receptor gamma levels followed a pattern closer to that of adipocyte number. Little change was observed in levels of transcripts for factors involved in de novo fatty acid synthesis, beta-oxidation, and lipolysis, suggesting that changes in the uptake of lipids from the circulation are the main mechanisms by which estrogen regulates lipid metabolism in these mice. PMID- 12639932 TI - Raloxifene acts as an estrogen agonist on the rabbit growth plate. AB - Estrogen treatment has been used to induce growth plate fusion, thereby reducing the final height in girls expected to achieve extreme tall stature. The treatment is effective, in terms of limiting final height, but concerns have been raised that it might also increase the risk for malignancies later in life. Raloxifene, a selective estrogen receptor modulator, has been shown to act as an estrogen agonist on bone density but as an estrogen antagonist on breast and uterine tissue. The effect of raloxifene treatment on growth plate fusion and final height is unknown. The aim of this study was to determine whether raloxifene would act as an estrogen agonist or antagonist on growth plate cartilage. Ovariectomized immature rabbits were treated for 4 wk with vehicle (controls), estradiol cypionate (E2), or raloxifene. Tibial growth velocity was decreased in both E2- (P < 0.001) and raloxifene-treated animals (P < 0.001), compared with controls. E2 and raloxifene treatment also decreased chondrocyte proliferation and the height of the proximal tibial growth plate. In addition, E2 and raloxifene hastened fusion of the distal tibial growth plate (P < 0.05) and decreased the number of proliferative and hypertrophic chondrocytes per column in the proximal tibial growth plate. As expected, the uterus was enlarged by estrogen, but not raloxifene, treatment. We conclude that raloxifene acts as an estrogen agonist on the growth plate, accelerating growth plate senescence and thus hastening epiphyseal fusion. PMID- 12639933 TI - Connection between integrins and cell activation in rat adrenal glomerulosa cells: a role for Arg-Gly-Asp peptide in the activation of the p42/p44(mapk) pathway and intracellular calcium. AB - Integrins are responsible for adhesion and activation of several intracellular cascades. The present study was aimed at determining whether the interaction between fibronectin and integrins could generate pathways involved in physiological functions of rat adrenal glomerulosa cells. Immunofluorescence studies and adhesion assays showed that fibronectin was the best matrix in promoting the formation of focal adhesion. Binding of glomerulosa cells to fibronectin, but not to collagen I or poly-L-lysine, involved the integrin binding sequence Arg-Gly-Asp (RGD). Activation of glomerulosa cells with Arg-Gly Asp-Ser (RGDS) induced an increase in [Ca(2+)](i), whereas fibronectin triggered a release of Ca(2+) from InsP(3)-sensitive Ca(2+) stores. Aldosterone secretion induced by ACTH, angiotensin II, and RGDS and proliferation were improved on fibronectin, compared with poly-L-lysine. The RGDS peptide induced a transient increase in the activity of the p42/p44(mapk), independent of phosphatidylinositol-3 kinase and protein kinase C. Integrins alpha(5) and alpha(V) as well as their fibronectin receptor partners beta(1) and beta(3), were identified. These results suggest that in rat adrenal glomerulosa cells, binding of the alpha(5)beta(1), alpha(v)beta(1), or alpha(v)beta(3) integrins to fibronectin is involved in the generation of two important signaling events, increase in intracellular calcium, and activation of the p42/p44(mapk) cascade, leading to cell proliferation and aldosterone secretion. PMID- 12639934 TI - Gonadotropin-releasing hormone attenuates pregnancy-associated thymic involution and modulates the expression of antiproliferative gene product prohibitin. AB - Thymic involution during pregnancy is believed to be a critical adaptive mechanism for regulation and control of the maternal immune system. These regulatory feedback mechanisms are important for the survival of the semiallogeneic fetus. In the present study, we examined the effects of GnRH on pregnancy-induced thymic involution by characterizing the expression patterns of prohibitin (PHB), an antiproliferative gene product, GnRH, and GnRH receptor (GnRH-R) proteins in the rat thymus and in mature splenic lymphocytes. GnRH agonist infusions in pregnant rats markedly attenuated pregnancy-induced thymic involution resulting in significant increases in thymic weight and thymocyte numbers. In addition, histological examination of the thymus revealed increase in cortical cellularity. Western blot analyses revealed a significant increase of total PHB protein content in thymi during pregnancy. Furthermore, distinct changes in PHB isoform expression were observed in the pregnant involuting thymi with greater expression of the basic PHB isoform. Basic isoform expression decreased in pregnant rats and was comparable with nonpregnant rat thymi upon GnRH agonist treatment. PHB is mainly expressed in mature cells of the thymic medulla, where it strongly colocalized with GnRH. We have observed GnRH-R immunoreactivity mainly in thymic medulla. Furthermore, as assessed by immunofluorescence double labeling with proliferating cell nuclear antigen, PHB was preferentially expressed in nonproliferating thymocytes. In this study, we demonstrated that GnRH, GnRH-R, and PHB show characteristic polarized expression in thymocytes. In addition, GnRH and PHB were coexpressed in mature splenic T cells. Our results suggest that PHB and GnRH are involved in thymic growth and may be important for maturation of T lymphocytes. PMID- 12639935 TI - Ghrelin-induced food intake is mediated via the orexin pathway. AB - The hypothalamus regulates energy intake by integrating the degree of starvation or satiation with the status of the environment through a variety of neuronal and blood-derived signals. Ghrelin, a peptide produced in the stomach and hypothalamus, stimulates feeding and GH secretion. Centrally administered ghrelin exerts an orexigenic activity through the neuropeptide Y (NPY) and agouti-related protein systems. The interaction between ghrelin and other hypothalamic orexigenic peptides, however, has not been clarified. Here, we investigated the anatomical interactions and functional relationship between ghrelin and two orexigenic peptides, orexin and melanin-concentrating hormone (MCH), present in the lateral hypothalamus. Ghrelin-immunoreactive axonal terminals made direct synaptic contacts with orexin-producing neurons. Intracerebroventricular administration of ghrelin induced Fos expression, a marker of neuronal activation, in orexin-producing neurons but not in MCH-producing neurons. Ghrelin remained competent to induce Fos expression in orexin-producing neurons following pretreatment with anti-NPY IgG. Pretreatment with anti-orexin-A IgG and anti orexin-B IgG, but not anti-MCH IgG, attenuated ghrelin-induced feeding. Administration of NPY receptor antagonist further attenuated ghrelin-induced feeding in rats treated with anti-orexin-IgGs. Ghrelin-induced feeding was also suppressed in orexin knockout mice. This study identifies a novel hypothalamic pathway that links ghrelin and orexin in the regulation of feeding behavior and energy homeostasis. PMID- 12639936 TI - Differential role of melanocortin receptor subtypes in cachexia. AB - Animals and humans respond to starvation with a complex neuroendocrine response that ultimately leads to an increase in appetite, a sparing of lean body mass (LBM) and burning of fat, and an overall decrease in basal metabolic rate. In contrast, cachexia is a pathological state of malnutrition associated with many infections and chronic diseases, wherein appetite is diminished concomitant with an increase in metabolic rate, and a relative wasting of LBM. In previous studies, we demonstrated that anorexia and weight loss in mouse cachexia models induced by lipopolysaccharide (LPS) administration and by tumor growth are ameliorated by central melanocortin-4 (MC4) receptor (MC4-R) blockade. In contrast to the results seen with MC4 blockade, melanocortin-3 (MC3) receptor knockout (MC3-RKO) mice show illness-induced anorexia and weight loss with LPS administration and with cytokine administration, and they have similar decreases in mobility. Both MC3-RKOs and MC4-RKOs have an intact corticosterone response and fever with LPS injection. In tumor models, we show that MC4-RKO mice resist the loss of LBM brought about by tumor growth, whereas MC3-RKO animals show enhanced tissue wasting. These data underscore the importance of central melanocortin signaling in weight homeostasis and demonstrate differential effects of MC3-R and MC4-R blockade on the development of cachexia. PMID- 12639938 TI - Aquaporin water channel genes are differentially expressed and regulated by ovarian steroids during the periimplantation period in the mouse. AB - The periimplantation period is marked by edematous changes in the uterus. In the mouse, increased uterine vascular permeability occurs in response to estrogen and certain vasoactive mediators, but the mechanisms that regulate fluid transport during implantation are not fully understood. Aquaporins (AQPs) are a family of membrane channel proteins that facilitate bulk water transport. To assess their role in implantation, we examined the expression of AQPs 0-9 in the mouse uterus on d 1-8 of pregnancy. Our results show distinct uterine expression patterns for AQP1, AQP4, and AQP5. AQP1 is localized to the inner circular myometrium throughout the periimplantation period. AQP4 is highly expressed in the luminal epithelium on d 1 of pregnancy but barely detectable at the time of implantation. AQP5 is expressed at low levels in the glandular epithelium during early pregnancy but is markedly increased on d 5. By immunohistochemistry, AQP5 is localized in the basolateral region of the uterine glands. Treatment of adult ovariectomized mice with replacement steroids demonstrates an estrogen-induced shift in AQP1 signals from the myometrium to the uterine stromal vasculature, suggesting a role in uterine fluid imbibition. In contrast, AQP5 is induced only in estrogen-treated, progesterone-primed uteri. We also observed expression of AQP8 in the inner-cell mass and AQP9 in the mural trophectoderm of the implanting blastocyst. Collectively, these results suggest that members of the AQP family are involved in embryo and uterine fluid homeostasis during implantation. PMID- 12639937 TI - Corticotropin-releasing factor receptor type 2 messenger ribonucleic acid in rat pituitary: localization and regulation by immune challenge, restraint stress, and glucocorticoids. AB - CRF receptor 2 (CRF R2) has been identified in the rat pituitary. However, the cell types that express the receptor remained to be determined. In the present study, we localized CRF R2 mRNA in gonadotropes of the anterior pituitary. Ribonuclease protection assays of anterior pituitary mRNA further showed that the dominant receptor type is CRF R2alpha. We also demonstrated that the expression of CRF R2 in the pituitary is sensitive to alterations to the hypothalamic pituitary-adrenal axis as CRF R2 mRNA levels in the anterior pituitary of male rats were significantly decreased 6 h after bacterial endotoxin lipopolysaccharide (LPS) injection or restraint stress. Subcutaneous corticosterone injections also resulted in significant suppression of CRF R2 mRNA levels in the pituitary, suggesting that glucocorticoids are involved in modulating CRF R2 mRNA levels in the pituitary under stress. LPS administration still caused a significant suppression of CRF R2 mRNA levels in the anterior pituitary of adrenalectomized rats. This suggests that one or more additional factors is involved in the regulation of CRF R2 expression in the anterior pituitary. Taken together, these data suggest that CRF R2alpha in the anterior pituitary might be involved in the regulation of gonadal functions under stress. PMID- 12639939 TI - Gonadotropin-releasing hormone neurons coexpress orexin 1 receptor immunoreactivity and receive direct contacts by orexin fibers. AB - The orexins are produced in neurons of the lateral hypothalamic area and implicated in the regulation of both feeding and reproductive function. Orexins stimulate LH secretion in steroid-primed ovariectomized female rats and suppress LH secretion in nonprimed ovariectomized rats. The aim of the present study was to characterize the neuroanatomical pathway by which orexin might modulate LH secretion in the rat. Using double- and triple-label immunofluorescence coupled with confocal microscopy, we found that 75-85% of GnRH neurons were contacted by orexin fibers, and triple labeling with synaptophysin provided additional confirmation of close contacts. Furthermore, about 85% of GnRH neurons were colocalized with the orexin receptor 1 (OX-R1), and the OX-R1-expressing GnRH neurons were contacted by orexin terminals, providing the basis for a functional neuroanatomical pathway. GnRH nerve terminals in the median eminence, however, do not express OX-R1. An additional study investigated the coexpression of neuropeptide Y Y4-like receptors and orexin fibers in relation to GnRH neurons. There is evidence that Y4 receptor stimulation results in LH release, and studies from our laboratory show Y4-like immunoreactivity in the majority of orexin cell bodies in the lateral hypothalamic area and some orexin fibers scattered throughout the hypothalamus. The present study found that, although Y4-positive orexin fibers are in present in the area of GnRH neurons, they never come in close contact with GnRH neurons. Together, these data suggest that Y4 receptor modulation of LH release is likely to be indirect through orexin cell bodies and that orexin modulates GnRH neurons directly via OX-R1. PMID- 12639940 TI - Rab8B GTPase and junction dynamics in the testis. AB - Throughout spermatogenesis, germ cells migrate from the basal to the adluminal compartment while remaining attached to Sertoli cells via actin-based adherens and intermediate filament-based anchoring junctions. However, the events that trigger deadhesion and adhesion remain largely unknown. As part of our continued effort in elucidating the mechanism of germ cell movement, we have examined the role of Rab8B, a GTPase probably participating in intracellular trafficking events at the site of the adherens junction. By RT-PCR Rab8B mRNA was found in the brain, testis, heart, kidney, and spleen. Immunohistochemical studies revealed that Rab8B was concentrated predominantly in the basal compartment, localizing to a similar site at which immunoreactive E-cadherin was found. Additional experiments demonstrated that Rab8B associated with the actin, intermediate filament, and microtubule cytoskeletal networks. When Sertoli cells were cultured at high density or germ cells were cocultured with Sertoli cells, Rab8B increased significantly during junction assembly. Moreover, inclusion of germ cell-conditioned medium in Sertoli cell cultures resulted in stimulation of Rab8B expression. Conversely, treatment of adult rats with 1-(2,4-dichlorobenzyl) indazole-3-carbohydrazide reduced Rab8B mRNA and protein levels, coinciding with the time of germ cell loss from the epithelium. Taken collectively, these studies suggest that Rab8B participates in adherens junction dynamics in the testis. PMID- 12639941 TI - Targeting sst2A receptor-expressing cells in the rat hypothalamus through in vivo agonist stimulation: neuroanatomical evidence for a major role of this subtype in mediating somatostatin functions. AB - Numerous physiological studies as well as in situ hybridization and PCR experiments concur in reporting a role for the sst2A receptor in transducing somatostatin (SRIF) actions in the rat hypothalamus. However, the distribution of this receptor protein is not known within this structure. Regional and cellular localization of the sst2A receptor was therefore examined in the rat hypothalamus using highly sensitive immunohistochemical techniques. In close correspondence with the distribution of SRIF-immunoreactive fibers, numerous hypothalamic areas displayed sst2A receptor immunoreactivity. Receptor labeling was, however, diffusely distributed over the tissue, and few immunopositive cells were apparent. Unraveling the distribution of receptor-expressing cells was achieved through acute in vivo agonist stimulation and subsequent receptor internalization. At the cellular level, double-immunolabeling experiments with synaptophysin and microtubule-associated protein 2 demonstrated that sst2A receptors were predominantly internalized in perikarya and dendrites. Double labeling experiments with SRIF revealed that 93% of arcuate, but only 18% of periventricular, SRIF-positive neurons expressed internalized receptors. Taken together, these results demonstrate for the first time that the sst2A receptor protein is widely, but selectively, distributed in the hypothalamus, and that postsynaptic sst2A auto- and heteroreceptors are well poised to play an important role in the somatostatinergic regulation of hypothalamic endocrine and metabolic processes. PMID- 12639942 TI - Somatostatin inhibits tumor angiogenesis and growth via somatostatin receptor-3 mediated regulation of endothelial nitric oxide synthase and mitogen-activated protein kinase activities. AB - Somatostatin was reported to inhibit Kaposi's sarcoma (KS) cell (KS-Imm) xenografts through an antiangiogenic activity. Here, we show that somatostatin blocks growth of established KS-Imm tumors with the same efficacy as adriamycin, a clinically effective cytotoxic drug. Whereas KS-Imm cells do not express somatostatin receptors (SSTRs), endothelial cells express several SSTRs, in particular SSTR3. We investigated the molecular mechanisms and receptor specificity of somatostatin inhibition of angiogenesis. Somatostatin significantly inhibited angiogenesis in vivo in the matrigel sponge assay; this inhibition was mimicked by the SSTR3 agonist L-796778 and reversed by the SSTR3 antagonist BN81658, demonstrating involvement of SSTR3. In vitro experiments showed that somatostatin directly affected different endothelial cell line proliferation through a block of growth-factor-stimulated MAPK and endothelial nitric oxide (NO) synthase (eNOS) activities. BN81658 reversed somatostatin inhibition of cell proliferation, NO production, and MAPK activity, indicating that SSTR3 activation is required for the effects of somatostatin in vitro. Finally in vivo angiogenesis assays demonstrated that eNOS inhibition was a prerequisite for the antiangiogenic effects of somatostatin, because high concentrations of sodium nitroprusside, an NO donor, abolished the somatostatin effects. In conclusion, we demonstrate that somatostatin is a powerful antitumor agent in vivo that inhibits tumor angiogenesis through SSTR3-mediated inhibition of both eNOS and MAPK activities. PMID- 12639943 TI - A new cell secreting insulin. AB - The pancreatic beta-cell is the only cell in animals that expresses the insulin gene and secretes insulin protein. We have found copious release of immunoreactive and bioactive insulin into the medium from the primary culture of carp adipocytes. Glucose augmented this release to more than 2-fold, and glucose transporter, Glut2, was detected in these cells. These all reflect characteristics of a pancreatic beta-cell. The expression of the adipocyte specific flotillin gene, the presence of peroxisomal proliferator-activated receptor gamma and Glut4, and the colocalization of insulin and leptin confirmed the identity of these cells as adipocytes. Purified carp adipocyte insulin (AdpInsl) comigrated with porcine and bovine insulin in SDS-PAGE, indicating the similarity of their molecular sizes (5.5 kDa). AdpInsl strongly reduced hyperglycemia in streptozotocin-induced diabetic rats. It also stimulated significantly higher glucose uptake in carp and hamster adipocytes than porcine insulin. Adipocyte RNA hybridized with rat and zebrafish insulin cDNA showing the expression of the insulin gene in this cell. Using oligonucleotide primers designed on the basis of conserved insulin domain, AdpInsl cDNA was reverse transcribed, cloned, and sequenced. The deduced amino acid sequence of AdpInsl A and B chain exhibited 98% homology with zebrafish and more than 70% homology with human, porcine, and murine insulin. To understand the structure-function relationship between AdpInsl and mammalian beta-cell insulin, we have analyzed the amino acid sequences and three-dimensional structure of AdpInsl. In the critical determinant segment for receptor binding, AdpInsl has His at the A8 position instead of Thr in human and porcine insulin, and this attributed greater biological activity to AdpInsl. Our results show that carp adipocyte is a unique cell. As an insulin target cell it can express the insulin gene and secrete highly active insulin protein; thus, it may serve as a natural alternative to pancreatic beta-cell insulin. PMID- 12639944 TI - Expression of ghrelin in the cyclic and pregnant rat ovary. AB - Ghrelin, a 28-amino acid acylated peptide, has been recently identified as the endogenous ligand for the GH secretagogue receptor. Previous studies demonstrated that ghrelin, acting centrally, strongly stimulates GH release and food intake. In this study we provide novel evidence for the expression of ghrelin in the cyclic and pregnant rat ovary. Persistent expression of ghrelin gene was demonstrated in rat ovary throughout the estrous cycle, although its relative mRNA levels varied depending on the stage of the cycle, with the lowest levels in proestrus and peak expression values on diestrous d 1, i.e. during the luteal phase of the cycle. Ghrelin immunoreactivity was predominantly located in the luteal compartment of the ovary; with intense immunostaining being detected in steroidogenic cells from corpus luteum of the current cycle as well as in all generations of regressing corpora lutea. Indeed, predominant expression of ghrelin in the corpus luteum was confirmed using a pseudopregnant rat model, where maximum ghrelin mRNA levels were detected in dissected luteal tissue. To note, the cyclicity in the profile of ovarian expression of ghrelin appeared to be tissue specific, as it was not detected in the stomach, nor was it observed in terms of circulating ghrelin levels. In addition, cyclic expression of ovarian ghrelin mRNA was disrupted by blockade of the preovulatory gonadotropin surge and ovulation by means of administration of a potent GnRH antagonist. Finally, ghrelin mRNA expression was persistently detected in rat ovary throughout pregnancy, with higher levels in early pregnancy and lower expression during the later part of gestation. In conclusion, our data provide novel evidence for the expression of ghrelin in the cyclic and pregnant rat ovary. Dynamic changes in the profile of ghrelin expression were detected during the estrous cycle and throughout pregnancy, thus suggesting a precise regulation of ovarian expression of ghrelin. Overall, our present findings may represent an additional link between body weight homeostasis and female reproductive function. PMID- 12639945 TI - Activin signaling through type IB activin receptor stimulates aromatase activity in the ovarian granulosa cell-like human granulosa (KGN) cells. AB - In addition to a stimulatory effect on FSH production by the pituitary gland, activin is thought to have a paracrine or autocrine role in follicular development in the ovary, where it is produced. Recently, we established a human ovarian granulosa tumor cell line, KGN, which possesses in vivo characteristics of granulosa cells, namely the expression of functional FSH receptors and cytochrome P-450 aromatase. Here, we have demonstrated the activin signaling pathway and its role in KGN cells. A series of transient transfection experiments revealed that activin type IB receptor (ActRIB) is an essential component of the activin signaling pathway in KGN cells. Smad2 was found to act downstream of ActRIB as an intracellular signal transmitter. Smad7, but not Smad6, was an inhibitory Smad in the pathway. Finally, we show that FSH receptor expression and cytochrome P-450 (P-450) aromatase activity was up-regulated by activin stimulation through ActRIB in KGN cells. These results show that we have clarified the signaling mechanisms and the roles of activin in the human granulosa cell line, KGN. Activin signaling mediated by ActRIB-Smad2 system in the ovary may thus be essential for the regulation of follicular differentiation. PMID- 12639946 TI - Androgen down-regulated and region-specific expression of germ cell nuclear factor in mouse epididymis. AB - Germ cell nuclear factor (GCNF), a nuclear orphan receptor, involved in spermatogenesis, neurogenesis, differentiation, and embryo development, was highly expressed with two transcripts (7.4 and 2.3 kb) in mouse testis and with only one transcript (7.4 kb) slightly expressed in brain, liver, and kidney. The 2.3-kb transcript was restricted to round spermatids at stages VII and VIII of the spermatogenic cycle. The present report demonstrated its expression in epididymis as well, but at a very low level. Northern blot analysis showed two transcripts: a common 7.4-kb transcript and a unique 3.1-kb transcript. The expression levels of both GCNF transcripts in epididymis were down-regulated by androgen, as observed in castrated animals and aged mice. Polyclonal antisera against GCNF protein were raised. Western blot analysis showed the presence of only one band in total protein extracts from either mouse testis or epididymis. It indicated that the two mRNAs (7.4 and 3.1 kb) encode for the same protein as in testis. Fluorescent immunohistochemical staining and in situ hybridization showed that its expression was in the principal cell abundant in the corpus region. It implies that some androgen-regulated gene expressions located at the corpus principal cells might be controlled by GCNF. PMID- 12639947 TI - Alphavbeta3 integrin signaling pathway is involved in insulin-like growth factor I-stimulated human extravillous trophoblast cell migration. AB - IGF-I and -II provide paracrine and autocrine stimuli, respectively, for extravillous trophoblast (EVT) cell migration. This study examined the role of alpha(v)beta(3) integrin and its signaling pathway in IGF-I-stimulated migration. Migration assays were conducted using cultured EVT cells treated with or without IGF-I in the presence or absence of alphaIR3, Arg-Gly-Asp (RGD) hexapeptide, and antibody against alpha(v)beta(3) integrin. Morphological changes were studied using scanning electron microscopy. Colocalization of alpha(5)beta(1) alpha(v)beta(3) integrins, vinculin, focal adhesion kinase, and paxillin were determined by immuno-cytochemistry and immunoblotting. The results showed that IGF-I could stimulate EVT cell migration in a time- and dose-dependent manner and addition of alphaIR3, Arg-Gly-Asp hexapeptide, and antibody against alpha(v)beta(3) integrin attenuated the IGF-I migratory effect. Scanning electron microscopy images revealed that IGF-I promoted lamellipodia formation. Immunostaining and immunoblotting exhibited the colocalization of alpha(v)beta(3) integrin with phosphorylated focal adhesion kinase, paxillin, and vinculin at focal adhesions after IGF-I treatment. Immunoblotting demonstrated an increase in focal adhesion kinase and paxillin tyrosine phosphorylation followed by tyrosine phosphorylation of IGF-I receptor in a time- and dose-dependent manner. These findings indicated alpha(v)beta(3) integrin localization in the core of focal adhesions of EVT cells and that alpha(v)beta(3) integrin signaling pathways are activated in IGF-I-mediated migration of these cells. PMID- 12639949 TI - Hypoxia-inducible factor and its biomedical relevance. PMID- 12639948 TI - Dual effect of pituitary adenylate cyclase activating polypeptide on prostate tumor LNCaP cells: short- and long-term exposure affect proliferation and neuroendocrine differentiation. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide that elicits the increase of intracellular cAMP levels and protein kinase A activity in various cell systems. Here we show that the pattern of cAMP elevation triggered by PACAP is critical for the fate of LNCaP prostate cancer cells. We demonstrate that these cells express PACAP and its type 1 receptor. A short-term stimulation with PACAP, which generates a transient cAMP rise, induces proliferation of LNCaP cells through a protein kinase A-dependent activation of the MAPK cascade. On the contrary, we observed that chronic PACAP stimulation, giving rise to a sustained cAMP accumulation, leads to proliferation arrest and neuroendocrine differentiation. Moreover, PACAP stimulates phosphory-lation and activation of the cAMP response element binding transcription factor (CREB), and MAPK activation is necessary for its full transcriptional activity, indicating a direct involvement of cAMP response element in PACAP action. These findings demonstrate that a crucial event determining the outcome of prostatic cancer cells progression is the sustained vs. transient intracellular cAMP increase. PMID- 12639950 TI - Escherichia coli YrbI is 3-deoxy-D-manno-octulosonate 8-phosphate phosphatase. AB - 3-Deoxy-d-manno-octulosonate 8-phosphate (KDO 8-P) phosphatase, which catalyzes the hydrolysis of KDO 8-P to KDO and inorganic phosphate, is the last enzyme in the KDO biosynthetic pathway for which the gene has not been identified. Wild type KDO 8-P phosphatase was purified from Escherichia coli B, and the N-terminal amino acid sequence matched a hypothetical protein encoded by the E. coli open reading frame, yrbI. The yrbI gene, which encodes for a protein of 188 amino acids, was cloned, and the gene product was overexpressed in E. coli. The recombinant enzyme is a tetramer and requires a divalent metal cofactor for activity. Optimal enzymatic activity is observed at pH 5.5. The enzyme is highly specific for KDO 8-P with an apparent K(m) of 75 microm and a k(cat) of 175 s(-1) in the presence of 1 mm Mg(2+). Amino acid sequence analysis indicates that KDO 8 P phosphatase is a member of the haloacid dehalogenase hydrolase superfamily. PMID- 12639952 TI - PAS domains. Common structure and common flexibility. AB - PAS (PER-ARNT-SIM) domains are a family of sensor protein domains involved in signal transduction in a wide range of organisms. Recent structural studies have revealed that these domains contain a structurally conserved alpha/beta-fold, whereas almost no conservation is observed at the amino acid sequence level. The photoactive yellow protein, a bacterial light sensor, has been proposed as the PAS structural prototype yet contains an N-terminal helix-turn-helix motif not found in other PAS domains. Here we describe the atomic resolution structure of a photoactive yellow protein deletion mutant lacking this motif, revealing that the PAS domain is indeed able to fold independently and is not affected by the removal of these residues. Computer simulations of currently known PAS domain structures reveal that these domains are not only structurally conserved but are also similar in their conformational flexibilities. The observed motions point to a possible common mechanism for communicating ligand binding/activation to downstream transducer proteins. PMID- 12639951 TI - MICoA, a novel metastasis-associated protein 1 (MTA1) interacting protein coactivator, regulates estrogen receptor-alpha transactivation functions. AB - The transcriptional activity of estrogen receptor-alpha (ER-alpha) is modified by coactivators, corepressors, and chromatin remodeling complexes. We have previously shown that the metastasis-associated protein-1 (MTA1), a component of histone deacetylase and nucleosome remodeling complexes, represses ER-driven transcription by recruiting histone deacetylases to the estrogen receptor element (ERE)-containing target gene chromatin in breast cancer cells. Using a yeast two hybrid screening to clone MTA1-interacting proteins, we identified a previously uncharacterized molecule, which we named as MTA1-interacting coactivator (MICoA). Our findings suggest that estrogen signaling promotes nuclear translocation of MICoA and that MICoA interacts with MTA1 both in vitro and in vivo. MICoA binds to the C-terminal region of MTA1, whereas MTA1 binds to the N-terminal MICoA containing one nuclear receptor interaction LSRLL motif. We showed that MICoA is an ER coactivator, cooperates with other ER coactivators, stimulates ER transactivation functions, and associates with the endogenous ER and its target gene promoter chromatin. MTA1 also repressed MICoA-mediated stimulation of ERE mediated transcription in the presence of ER and ER variants with naturally occurring mutations, such as D351Y and K303R, and that it interfered with the association of MICoA with the ER-target gene chromatin. Because chromatin is a highly dynamic structure and because MTA1 and MICoA could be detected within the same complex, these findings suggest that MTA1 and MICoA might transmodulate functions of each other and any potential deregulation of MTA1 is likely to contribute to the functional inactivation of the ER pathway, presumably by derecruitment of MICoA from ER target promoter chromatin. PMID- 12639953 TI - Exosome release is regulated by a calcium-dependent mechanism in K562 cells. AB - Multivesicular bodies (MVBs) are endocytic structures that contain small vesicles formed by the budding of an endosomal membrane into the lumen of the compartment. Fusion of MVBs with the plasma membrane results in secretion of the small internal vesicles termed exosomes. K562 cells are a hematopoietic cell line that releases exosomes. The application of monensin (MON) generated large MVBs that were labeled with a fluorescent lipid. Exosome release was markedly enhanced by MON treatment, a Na+/H+ exchanger that induces changes in intracellular calcium (Ca2+). To explore the possibility that the effect of MON on exosome release was caused via an increase in Ca2+, we have used a calcium ionophore and a chelator of intracellular Ca2+. Our results indicate that increasing intracellular Ca2+ stimulates exosome secretion. Furthermore, MON-stimulated exosome release was completely eliminated by 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester (BAPTA-AM), implying a requirement for Ca2+ in this process. We have observed that the large MVBs generated in the presence of MON accumulated Ca2+ as determined by labeling with Fluo3-AM, suggesting that intralumenal Ca2+ might play a critical role in the secretory process. Interestingly, our results indicate that transferrin (Tf) stimulated exosome release in a Ca2+-dependent manner, suggesting that Tf might be a physiological stimulus for exosome release in K562 cells. PMID- 12639954 TI - Characteristics of the interaction of a synthetic human tristetraprolin tandem zinc finger peptide with AU-rich element-containing RNA substrates. AB - Tristetraprolin (TTP) and its two known mammalian family members are tandem CCCH zinc finger proteins that can bind to AU-rich elements (AREs) in cellular mRNAs and destabilize those transcripts, apparently by initiating their deadenylation. Previous studies have shown that the approximately 70-amino acid tandem zinc finger domain of TTP is required and sufficient for RNA binding, and that the integrity of both zinc fingers is also required. However, little is known about the kinetics or structure of the peptide-RNA interaction, in part because of difficulties in obtaining soluble recombinant protein or peptides. We characterized the binding of a synthetic 73-amino acid peptide from human TTP to the tumor necrosis factor (TNF) ARE by gel mobility shift analyses and fluorescence anisotropy experiments. Both types of studies yielded a peptide-RNA dissociation constant of approximately 10 nM. Surprisingly, we found that the "footprint" from the TNF ARE required for peptide binding was only approximately 9 bases and that two molecules of peptide could bind to probes containing as little as 19 bases. An identical recombinant peptide exhibited gel shift characteristics similar to those of the synthetic peptide. NMR analysis of the 15N-labeled recombinant peptide suggested that its first zinc finger was structured in solution but that the second was not. The titration of oligonucleotides representing 17, 13, and even 9 bases of the TNF ARE caused an essentially identical, dramatic shift of existing resonances, and the appearance of new resonances in the peptide spectra, so that all amino acids could be assigned. These data suggest that this TTP peptide-RNA complex is structured in solution and might be amenable to NMR structure determination. PMID- 12639955 TI - Thermosensor action of GrpE. The DnaK chaperone system at heat shock temperatures. AB - Temperature directly controls functional properties of the DnaK/DnaJ/GrpE chaperone system. The rate of the high to low affinity conversion of DnaK shows a non-Arrhenius temperature dependence and above approximately 40 degrees C even decreases. In the same temperature range, the ADP/ATP exchange factor GrpE undergoes an extensive, fully reversible thermal transition (Grimshaw, J. P. A., Jelesarov, I., Schonfeld, H. J., and Christen, P. (2001) J. Biol. Chem. 276, 6098 6104). To show that this transition underlies the thermal regulation of the chaperone system, we introduced an intersubunit disulfide bond into the paired long helices of the GrpE dimer. The transition was absent in disulfide-linked GrpE R40C but was restored by reduction. With disulfide-stabilized GrpE, the rate of ADP/ATP exchange and conversion of DnaK from its ADP-liganded high affinity R state to the ATP-liganded low affinity T state continuously increased with increasing temperature. With reduced GrpE R40C, the conversion became slower at temperatures >40 degrees C, as observed with wild-type GrpE. Thus, the long helix pair in the GrpE dimer acts as a thermosensor that, by decreasing its ADP/ATP exchange activity, induces a shift of the DnaK.substrate complexes toward the high affinity R state and in this way adapts the DnaK/DnaJ/GrpE system to heat shock conditions. PMID- 12639956 TI - Crystal structures of allosamidin derivatives in complex with human macrophage chitinase. AB - The pseudotrisaccharide allosamidin is a potent family 18 chitinase inhibitor with demonstrated biological activity against insects, fungi, and the Plasmodium falciparum life cycle. The synthesis and biological properties of several derivatives have been reported. The structural interactions of allosamidin with several family 18 chitinases have been determined by x-ray crystallography previously. Here, a high resolution structure of chitotriosidase, the human macrophage chitinase, in complex with allosamidin is presented. In addition, complexes of the allosamidin derivatives demethylallosamidin, methylallosamidin, and glucoallosamidin B are described, together with their inhibitory properties. Similar to other chitinases, inhibition of the human chitinase by allosamidin derivatives lacking a methyl group is 10-fold stronger, and smaller effects are observed for the methyl and C3 epimer derivatives. The structures explain the effects on inhibition in terms of altered hydrogen bonding and hydrophobic interactions, together with displaced water molecules. The data reported here represent a first step toward structure-based design of specific allosamidin derivatives. PMID- 12639957 TI - The cationic amphipathic alpha-helix of HIV-1 viral protein R (Vpr) binds to nucleic acids, permeabilizes membranes, and efficiently transfects cells. AB - Viral protein R (Vpr) is a small protein of 96 amino acids that is conserved among the lentiviruses human immunodeficiency virus type 1 (HIV-1), HIV-2, and simian immunodeficiency virus. We recently sought to determine whether the karyophilic properties of Vpr, as well as its ability to bind nucleic acids, could be used to deliver DNA into cells. We have found that the C-terminal domain of Vpr-(52-96) is able to efficiently transfect various cell lines. Here, we show that the shortest active sequence for gene transfer corresponds to the domain that adopts a alpha-helix conformation. DNA binding studies and permeabilization assays performed on cells demonstrated that the peptides that are efficient in transfection condense plasmid DNA and are membranolytic. Electron microscopy studies and transfection experiments performed in the presence of inhibitors of the endocytic processes indicated that the major entry pathway of Vpr-DNA complexes is through endocytosis. Taken together, the results show that the cationic C-terminal alpha-helix of Vpr has DNA-condensing as well as membrane destabilizing capabilities, both properties that are indispensable for efficient DNA transfection. PMID- 12639958 TI - Membrane topology of gamma-secretase component PEN-2. AB - PEN-2 is an integral membrane protein that is a necessary component of the gamma secretase complex, which is central in the pathogenesis of Alzheimer's disease and is also required for Notch signaling. In the absence of PEN-2, Notch signaling fails to guide normal development in Caenorhabditis elegans, and amyloid beta peptide is not generated from the amyloid precursor protein. Human PEN-2 is a 101-amino acid protein containing two putative transmembrane domains. To understand its interaction with other gamma-secretase components, it is important to know the membrane topology of each member of the complex. To characterize the membrane topology of PEN-2, we introduced single amino acid changes in each of the three hydrophilic regions of PEN-2 to generate N-linked glycosylation sites. We found that the N-linked glycosylation sites present in the N- and C-terminal domains of PEN-2 were utilized, whereas a site in the hydrophilic "loop" region connecting the two transmembrane domains was not. The addition of a carbohydrate structure in the N-terminal domain of PEN-2 prevented association with presenilin 1, whereas glycosylation in the C-terminal region of PEN-2 did not, suggesting that the N-terminal domain is important for interactions with presenilin 1. Immunofluorescence microscopy with selective permeabilization of the plasma membrane of cells expressing epitope-tagged forms of PEN-2 confirmed the lumenal location of both the N and C termini. A protease protection assay also demonstrated that the loop domain of PEN-2 is cytosolic. Thus, PEN-2 spans the membrane twice, with the N and C termini facing the lumen of the endoplasmic reticulum. PMID- 12639959 TI - Assessing the role of the T cell receptor beta gene enhancer in regulating coding joint formation during V(D)J recombination. AB - To assess the role of the T cell receptor (TCR) beta gene enhancer (Ebeta) in regulating the processing of VDJ recombinase-generated coding ends, we assayed TCRbeta rearrangement of Ebeta-deleted (DeltaEbeta) thymocytes in which cell death is inhibited via expression of a Bcl-2 transgene. Compared with DeltaEbeta, DeltaEbeta Bcl-2 thymocytes show a small accumulation of TCRbeta standard recombination products, including coding ends, that involves the proximal Dbeta Jbeta and Vbeta14 loci but not the distal 5' Vbeta genes. These effects are detectable in double negative pro-T cells, predominate in double positive pre-T cells, and correlate with regional changes in chromosomal structure during double negative-to-double positive differentiation. We propose that Ebeta, by driving long range nucleoprotein interactions and the control of locus expression and chromatin structure, indirectly contributes to the stabilization of coding ends within the recombination processing complexes. The results also illustrate Ebeta dependent and -independent changes in chromosomal structure, suggesting distinct modes of regulation of TCRbeta allelic exclusion depending on the position within the locus. PMID- 12639960 TI - Microarray analysis reveals interleukin-6 as a novel secretory product of the hypothalamo-neurohypophyseal system. AB - Physiological activation of the hypothalamo-neurohypophyseal system (HNS) by dehydration results is a massive release of vasopressin (VP) from the posterior pituitary. This is accompanied by a functional remodeling of the HNS. In this study we used cDNA arrays in an attempt to identify genes that exhibit differential expression in the hypothalamus following dehydration. Our study revealed nine candidate genes, including interleukin-6 (IL-6) as a putative novel secretory product of HNS worthy of further analysis. In situ hybridization and immunocytochemistry confirmed that IL-6 is robustly expressed in the supraoptic (SON) and the paraventricular (PVN) nuclei of the hypothalamus. By double staining immunofluorescence we showed that IL-6 is largely co-localized with VP in the SON and PVN. In situ hybridization, immunocytochemistry, and Western blotting all revealed IL-6 up-regulation in the SON and PVN following dehydration, thus validating the array data. The same dehydration stimulus resulted in an increase in IL-6 immunoreactivity in the axons of the internal zone of the median eminence and a marked reduction in IL-6-like material in the posterior pituitary gland. We thus suggest that IL-6 takes the same secretory pathway as VP and is secreted from the posterior pituitary following a physiological stimulus. PMID- 12639961 TI - Differential effects of scavenger receptor BI deficiency on lipid metabolism in cells of the arterial wall and in the liver. AB - Scavenger receptor class B, type I (SRBI) is a key regulator of high density lipoprotein (HDL) metabolism. It facilitates the efflux of cholesterol from cells in peripheral tissues to HDL and mediates the selective uptake of cholesteryl esters from HDL in the liver. We investigated the effects of SRBI deficiency in the arterial wall and in the liver using SRBI-deficient mice and wild-type littermates fed a Western-type diet. The SRBI-deficient mice showed massive accumulation of cholesterol-rich HDL in the circulation, reflecting impaired delivery to the liver. Strikingly, SRBI deficiency did not alter hepatic cholesterol (ester) content nor did it affect the expression of key regulators of hepatic cholesterol homeostasis, including HMG-CoA reductase, the low density lipoprotein receptor, and cholesterol 7alpha-hydroxylase. However, a approximately 40% reduction in biliary cholesterol content was observed, and the expression of ABCG8 and ABCG5, ATP half-transporters implicated in the transport of sterols from the liver to the bile, was attenuated by 70 and 35%, respectively. In contrast to the situation in the liver, SRBI deficiency did result in lipid deposition in the aorta and atherosclerosis. Vascular mRNA analysis showed increased expression of inflammatory markers as well as of genes involved in cellular cholesterol homeostasis. Our data show that, although hepatic cholesterol homeostasis is maintained upon feeding a Western-type diet, SRBI deficiency is associated with de-regulation of cholesterol homeostasis in the arterial wall that results in an increased susceptibility to atherosclerosis. PMID- 12639963 TI - The prostate-derived sterile 20-like kinase (PSK) regulates microtubule organization and stability. AB - Sterile 20 (STE20) protein kinases, which include germinal center kinases and p21 activated protein kinases, are known to activate mitogen-activated protein kinase pathways (c-Jun NH(2)-terminal kinase, p38, or extracellular signal-regulated kinase), leading to changes in gene transcription. Some STE20s can also regulate the cytoskeleton, and we have shown that the germinal center kinase-like kinase prostate-derived STE20-like kinase (PSK) affects actin cytoskeletal organization. Here, we demonstrate that PSK colocalizes with microtubules; and that this localization is disrupted by the microtubule depolymerizing agent nocodazole. The association of PSK with microtubules results in the production of stabilized perinuclear microtubule cables that are nocodazole-resistant and contain increased levels of acetylated alpha-tubulin. Kinase-defective PSK (K57A) or the C terminus of PSK (amino acids 745-1235) lacking the kinase domain are sufficient for microtubule binding and stabilization, demonstrating that the catalytic activity of the protein is not required. The localization of PSK to microtubules occurs via its C terminus, and PSK binds and phosphorylates alpha- and beta tubulin in vitro. The N terminus of PSK (1-940) is unable to bind or stabilize microtubules, demonstrating that PSK must associate with microtubules for their reorganization to occur. These results demonstrate that PSK interacts with microtubules and affects their organization and stability independently of PSK kinase activity. PMID- 12639962 TI - The hepatitis C virus core protein modulates T cell responses by inducing spontaneous and altering T-cell receptor-triggered Ca2+ oscillations. AB - Alterations of cytokine responses are thought to favor the establishment of persistent hepatitis C virus (HCV) infection, enhancing the risk of liver cirrhosis and hepatocellular carcinoma. Expression of the HCV core (C) protein modulates transcription of the IL-2 promoter in T lymphocytes by activating the nuclear factor of activated T lymphocyte (NFAT) pathway. Here we report on the effect of HCV C on Ca2+ signaling, which is essential for activation of NFAT. Expression of HCV C correlated with increased levels of cytosolic Ca2+ and spontaneous Ca2+ oscillations in transfected Jurkat cells. Triggering of the T cell receptor induced a prolonged Ca2+ response characterized by vigorous high frequent oscillations in a high proportion of the responding cells. This was associated with decreased sizes and accelerated emptying of the intracellular calcium stores. The effect of HCV C on calcium mobilization was not dependent on phospholipase C-gamma 1 (PLC-gamma) activity or increased inositol 1,4,5 trisphosphate (IP3) production and did not require functional IP3 receptors, suggesting that insertion of the viral protein in the endoplasmic reticulum membrane may be sufficient to promote Ca2+ leakage with dramatic downstream consequences on the magnitude and duration of the response. Our data suggest that expression of HCV C in infected T lymphocytes may contribute to the establishment of persistent infections by inducing Ca2+ oscillations that regulate both the efficacy and information content of Ca2+ signals and are ultimately responsible for induction of gene expression and functional differentiation. PMID- 12639964 TI - Common location of determinants in initiator transfer RNAs for initiator elongator discrimination in bacteria and in eukaryotes. AB - Initiator tRNAs are used exclusively for initiation of protein synthesis and not for elongation. We show that both Escherichia coli and eukaryotic initiator tRNAs have negative determinants, at the same positions, that block their activity in elongation. The primary negative determinant in E. coli initiator tRNA is the C1xA72 mismatch at the end of the acceptor stem. The primary negative determinant in eukaryotic initiator tRNAs is located in the TPsiC stem, whereas a secondary negative determinant is the A1:U72 base pair at the end of the acceptor stem. Here we show that E. coli initiator tRNA also has a secondary negative determinant for elongation and that it is the U50.G64 wobble base pair, located at the same position in the TPsiC stem as the primary negative determinant in eukaryotic initiator tRNAs. Mutation of the U50.G64 wobble base pair to C50:G64 or U50:A64 base pairs increases the in vivo amber suppressor activity of initiator tRNA mutants that have changes in the acceptor stem and in the anticodon sequence necessary for amber suppressor activity. Binding assays of the mutant aminoacyl-tRNAs carrying the C50 and A64 changes to the elongation factor EF-Tu.GTP show marginally higher affinity of the C50 and A64 mutant tRNAs and increased stability of the EF-Tu.GTP. aminoacyl-tRNA ternary complexes. Other results show a large effect of the amino acid attached to a tRNA, glutamine versus methionine, on the binding affinity toward EF-Tu.GTP and on the stability of the EF-Tu.GTP.aminoacyl-tRNA ternary complex. PMID- 12639966 TI - Stk10, a new member of the polo-like kinase kinase family highly expressed in hematopoietic tissue. AB - The Ste20 family of serine/threonine kinases plays an important role in numerous cellular functions such as growth, apoptosis, and morphogenesis. We have identified a previously cloned but uncharacterized family member termed Stk10, which is a human homolog of murine Lok, a serine/threonine kinase highly expressed in lymphocytes. Northern analysis demonstrated that the Stk10 transcript is present in many tissues, although highest expression levels are seen in hematopoietic cells. Due to close sequence homology to human Slk and Xenopus laevis xPlkk1, two polo-like kinase kinases, we investigated whether Stk10 might also play a role as a Plk1 activator. Plk1 has been shown to be overexpressed in multiple tumor types, thus attracting high interest to its potential upstream regulators. We show here that Stk10 can associate with Plk1 in cells and furthermore can phosphorylate Plk1 in vitro. Engineered NIH-3T3 cell lines that overexpress a dominant negative version of Stk10 display an altered cell cycle phenotype characterized by increased DNA content, raising the possibility that expression of a dominant negative Stk10 may impinge upon Plk1 function in vivo; it has previously been shown that unregulated expression of Plk1 can result in a variety of nuclear defects. We suggest, therefore, that Stk10 is a novel polo-like kinase kinase that cooperates with hSlk to regulate Plk1 function in human cells. PMID- 12639965 TI - Thymidine phosphorylase and 2-deoxyribose stimulate human endothelial cell migration by specific activation of the integrins alpha 5 beta 1 and alpha V beta 3. AB - Thymidine phosphorylase is an angiogenic factor that is frequently overexpressed in solid tumors, in rheumatoid arthritis, and in response to inflammatory cytokines. Our previous studies showed that cells expressing thymidine phosphorylase stimulated endothelial cell migration in vitro. This was a consequence of the intracellular metabolism of thymidine by thymidine phosphorylase and subsequent extracellular release of 2-deoxyribose. The mechanisms by which 2-deoxyribose might mediate thymidine phosphorylase-induced cell migration in vitro, however, are obscure. Here we show that both thymidine phosphorylase and 2-deoxyribose stimulated the formation of focal adhesions and the tyrosine 397 phosphorylation of focal adhesion kinase in human umbilical vein endothelial cells. Although similar actions occurred upon treatment with the angiogenic factor vascular endothelial growth factor (VEGF), thymidine phosphorylase differed from VEGF in that its effect on endothelial cell migration was blocked by antibodies to either integrin alpha 5 beta 1 or alpha v beta 3, whereas VEGF-induced endothelial cell migration was only blocked by the alpha v beta 3 antibody. Further, thymidine phosphorylase and 2-deoxyribose, but not VEGF, increased the association of both focal adhesion kinase and the focal adhesion-associated protein vinculin with integrin alpha 5 beta 1 and, in intact cells, increased the co-localization of focal adhesion kinase with alpha 5 beta 1. Thymidine phosphorylase and 2-deoxyribose-induced focal adhesion kinase phosphorylation was blocked by the antibodies to alpha 5 beta 1 and alpha v beta 3, directly linking the migration and signaling components of thymidine phosphorylase and 2-deoxyribose action. Cell surface expression of alpha 5 beta 1 was also increased by thymidine phosphorylase and 2-deoxyribose. These experiments are the first to demonstrate a direct effect of thymidine phosphorylase and 2-deoxyribose on signaling pathways associated with endothelial cell migration. PMID- 12639967 TI - Regulation of the p53 homolog p73 by adenoviral oncogene E1A. AB - p73 is a p53 homolog, as they are similar structurally and functionally. Unlike p53, p73 is not inactivated by the products of viral oncogenes such as SV40 T antigen and human papilloma virus E6. Here we show that the product of adenoviral oncogene E1A inhibits the transcriptional activation by both p73alpha and p73beta. Electrophoretic mobility shift assays revealed that E1A does not inhibit the sequence-specific DNA binding by p73. Transcriptional activation by a fusion protein containing the Gal4 DNA-binding domain and either of the activation domains of p73 was inhibited by wild-type (WT) E1A, but not by the N-terminal deletion mutant E1A(Delta2-36). E1A(Delta2-36), which does not bind to the p300/CBP family of coactivators, failed to inhibit p73-mediated transcription, whereas E1A(DeltaCR2), a deletion mutant that does not bind to the pRb family of proteins, inhibited p73-mediated transcription as efficiently as WT E1A. Consistent with these observations, growth arrest induced by p73 expressed from a recombinant adenovirus was abrogated by WT E1A, which correlated with inhibition of p73-mediated induction of p21(WAF1/CIP1) by E1A. However, p73 was able to induce p21(WAF1/CIP1) and to mediate growth arrest in the presence of E1A(Delta2 36). Furthermore, the expression of either wild-type E1A or E1A(Delta2-36) resulted in the stabilization of endogenous p73. However, p73 stabilized in response to the expression of E1A(Delta2-36), but not WT E1A, was able to activate the expression of p21(WAF1/CIP1). These results suggest that the transcriptional activation function of p73 is specifically targeted by E1A through a mechanism involving p300/CBP proteins during the process of transformation and that p73 may have a role to play as a tumor suppressor. PMID- 12639968 TI - Observed hysteresis of virus capsid disassembly is implicit in kinetic models of assembly. AB - For many protein multimers, association and dissociation reactions fail to reach the same end point; there is hysteresis preventing one and/or the other reaction from equilibrating. We have studied in vitro assembly of dimeric hepatitis B virus (HBV) capsid protein and dissociation of the resulting T = 4 icosahedral capsids. Empty HBV capsids composed of 120 capsid protein dimers were more resistant to dissociation by dilution or denaturants than anticipated from assembly experiments. Using intrinsic fluorescence, circular dichroism, and size exclusion chromatography, we showed that denaturants dissociate the HBV capsids without unfolding the capsid protein; unfolding of dimer only occurred at higher denaturant concentrations. The apparent energy of interaction between dimers measured in dissociation experiments was much stronger than when measured in assembly studies. Unlike assembly, capsid dissociation did not have the concentration dependence expected for a 120-subunit complex; consequently the apparent association energy systematically varied with reactant concentration. These data are evidence of hysteresis for HBV capsid dissociation. Simulations of capsid assembly and dissociation reactions recapitulate and provide an explanation for the observed behavior; these results are also applicable to oligomeric and multidomain proteins. In our calculations, we find that dissociation is impeded by temporally elevated concentrations of intermediates; this has the paradoxical effect of favoring re-assembly of those intermediates despite the global trend toward dissociation. Hysteresis masks all but the most dramatic decreases in contact energy. In contrast, assembly reactions rapidly approach equilibrium. These results provide the first rigorous explanation of how virus capsids can remain intact under extreme conditions but are still capable of "breathing." A biological implication of enhanced stability is that a triggering event may be required to initiate virus uncoating. PMID- 12639969 TI - Excision of the first intron from the gonadotropin-releasing hormone (GnRH) transcript serves as a key regulatory step for GnRH biosynthesis. AB - The mammalian gonadotropin-releasing hormone (GnRH) gene consists of four short exons (denoted as 1, 2, 3, and 4) and three intervening introns (A, B, and C). Recently, we demonstrated that excision of the first intron (intron A) from the GnRH transcript is regulated in a tissue- and developmental stage-specific fashion and is severely attenuated in hypogonadal (hpg) mouse because of its lack of exonic splicing enhancers (ESE) 3 and 4. In the present study, we examined the influence of intron A on translational efficiency, thereby establishing a post transcriptional control over GnRH biosynthesis. First, we verified that an intron A-retained GnRH transcript is a splicing variant but not a splicing intermediate. Intron A-retained transcripts can be transported to the cytoplasm in contrast to intron B-containing transcripts, which are restricted to the nucleus. This result implicates the intron A-retained GnRH transcript as a splicing variant; it has a long 5'-untranslated region, as the GnRH prohormone open reading frame (ORF) begins on exon 2. We investigated whether an intron A-retained GnRH transcript can properly initiate translation at the appropriate start codon and found that intron A completely blocks the translation initiation of its downstream reporter ORF both in vivo and in vitro. The inhibition of translation initiation appears to be due to the presence of a tandem repeat of ATG sequences within intron A. Constructs bearing mutations of ATGs to AAGs restored translation initiation at the downstream start codon; the extent of this restoration correlated with the number of mutated ATGs. Besides the failure in the translation initiation of GnRH coding region in the intron A-containing variant, the present study also suggests that the interference between mature GnRH mRNA and intron A-retained splicing variant could occur to lower the efficiency of GnRH biosynthesis in the GT1-1 immortalized GnRH-producing cell line. Therefore, our results indicate that the precise and efficient excision of intron A and the joining of adjacent exons may be a critical regulatory step for the post-transcriptional regulation of GnRH biosynthesis. PMID- 12639970 TI - The neuroprotective effects of heat shock protein 27 overexpression in transgenic animals against kainate-induced seizures and hippocampal cell death. AB - The 27-kDa heat shock protein (HSP27) has a potent ability to increase cell survival in response to a wide range of cellular challenges. In order to investigate the mode of action of HSP27 in vivo, we have developed transgenic lines, which express human HSP27 at high levels throughout the brain, spinal cord, and other tissues. In view of the particular property of HSP27 compared with other HSPs to protect neurons against apoptosis, we have tested these transgenic lines in a well established in vivo model of neurotoxicity produced by kainic acid, where apoptotic cell death occurs. Our results demonstrate for the first time the marked protective effects of HSP27 overexpression in vivo, which significantly reduces kainate-induced seizure severity and mortality rate (>50%) in two independent lines and markedly reduces neuronal cell death in the CA3 region of hippocampus. This reduced seizure severity in HSP27 transgenic animals was associated with a marked attenuation of caspase 3 induction and apoptotic features. These studies clearly demonstrate that HSP27 has a major neuroprotective effect in the central nervous system in keeping with its properties demonstrated in culture and highlight an early stage in the cell death pathway that is affected by HSP27. PMID- 12639971 TI - Glucuronidation of arachidonic and linoleic acid metabolites by human UDP glucuronosyltransferases. AB - Arachidonic acids (AA) and linoleic acids (LAs) are metabolized, in several tissues, to hydroxylated metabolites that are important mediators of many physiological and pathophysiological processes. The conjugation of leukotriene B4 (LTB4), 5-hydroxyeicosatetraenoic acid (HETE), 12-HETE, 15-HETE, and 13 hydroxyoctadecadienoic acid (HODE) by the human UDP-glucuronosyltransferase (UGT) enzymes was investigated. All substrates tested were efficiently conjugated by human liver microsomes to polar derivatives containing the glucuronyl moiety as assessed by mass spectrometry. The screening analyses with stably expressed UGT enzymes in HK293 showed that glucuronidation of LTB4 was observed with UGT1A1, UGT1A3, UGT1A8, and UGT2B7, whereas UGT1A1, UGT1A3, UGT1A4, and UGT1A9 also conjugated most of the HETEs and 13-HODE. LA and AA metabolites also appear to be good substrates for the UGT2B subfamily members, especially for UGT2B4 and UGT2B7 that conjugate all HETE and 13-HODE. Interestingly, UGT2B10 and UGT2B11, which are considered as orphan enzymes since no conjugation activity has so far been demonstrated with these enzymes, conjugated 12-HETE, 15-HETE, and 13-HODE. In summary, our data showed that several members of UGT1A and UGT2B families are capable of converting LA and AA metabolites into glucuronide derivatives, which is considered an irreversible step to inactivation and elimination of endogenous substances from the body. PMID- 12639972 TI - Fatty acid flux suppresses fatty acid synthesis in hamster intestine independently of SREBP-1 expression. AB - Hamsters were fed a control diet or diets containing palm, olive, safflower, or fish oil for 2 weeks. In villus cell populations from duodenum, jejunum, and ileum, rates of intestinal fatty acid and cholesterol synthesis were estimated, as were sterol regulatory element-binding protein (SREBP)-1a, SREBP-1c, SREBP-2, HMG-CoA synthase, fatty acid synthase, ATP citrate lyase, acetyl-CoA carboxylase mRNA levels, and SREBP-1 and SREBP-2 mass. Plasma cholesterol and triacylglcerol levels were increased in animals ingesting palm oil and decreased in animals ingesting fish oil. Fatty acid synthesis and fatty acid synthase activity were decreased in the proximal intestine of animals ingesting all the fat-containing diets. Intestinal cholesterol synthesis was unaltered. In animals fed fat, SREBP 1c gene expression was modestly increased in the duodenum of hamsters fed palm oil or olive oil, and decreased in animals ingesting safflower oil or fish oil. Fatty acid synthase, acetyl-CoA carboxylase, ATP citrate lyase, SREBP-2, and HMG CoA synthase mRNA levels were not altered, nor were SREBP-1 or SREBP-2 mass. In the intestine, dietary polyunsaturated fatty acids suppress SREBP-1c mRNA without altering expression of its target genes, fatty acid synthase, acetyl-CoA carboxylase, or ATP citrate lyase. Fatty acid influx decreases intestinal fatty acid synthesis by a posttranscriptional mechanism independent of the SREBP pathway. PMID- 12639973 TI - Visualizing caveolin-1 and HDL in cholesterol-loaded aortic endothelial cells. AB - Caveolae are vesicular invaginations of the plasma membranes that regulate signal transduction and transcytosis, as well as cellular cholesterol homeostasis. Our previous studies indicated that the removal of cholesterol from aortic endothelial cells and smooth muscle cells in the presence of HDL is associated with plasmalemmal invaginations and plasmalemmal vesicles. The goal of the present study was to investigate the location and distribution of caveolin-1, the main structural protein component of caveolae, in cholesterol-loaded aortic endothelial cells after HDL incubation. Confocal microscopic analysis demonstrated that the caveolin-1 appeared to colocalize with HDL-fluorescein 1,1' dioctadecyl 3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) conjugates on the cell surface. No free HDL-DiI conjugates were revealed in the cytoplasm. Immunoelectron microscopy further demonstrated that caveolin-1 gold (15 nm) conjugates colocalized with HDL gold (10 nm) conjugates in the plasmalemmal invaginations. These morphological results indicated that caveolae are the major membrane domains facilitating the transport of excess cholesterol to HDL on the cell surface of aortic endothelial cells. PMID- 12639974 TI - Hepatic lipase and dyslipidemia: interactions among genetic variants, obesity, gender, and diet. AB - Hepatic lipase (HL) plays a central role in LDL and HDL remodeling. High HL activity is associated with small, dense LDL particles and with reduced HDL2 cholesterol levels. HL activity is determined by an HL gene promoter polymorphism, by gender (lower in premenopausal women), and by visceral obesity with insulin resistance. The activity is affected by dietary fat intake and selected medications. There is evidence for an interaction of the HL promoter polymorphism with visceral obesity, dietary fat intake, and with lipid-lowering medications in determining the level of HL activity. The dyslipidemia with high HL activity is a potentially proatherogenic lipoprotein profile in the metabolic syndrome, in Type 2 diabetes, and in familial combined hyperlipidemia. PMID- 12639975 TI - Natural genetic variation as a tool in understanding the role of CETP in lipid levels and disease. AB - Since the identification of cholesteryl ester transfer protein (CETP), its role in the modulation of HDL levels and cardiovascular disease has been debated. With the early detection of genetic variants followed by the finding of families deficient in CETP, genetic studies have played a large role in the attempts to understand the association of CETP with lipids and disease; however, results of these studies have often led to disparate conclusions. With the availability of a greater variety of genetic polymorphisms and larger studies in which disease has been examined, it is now possible to compare the breadth of CETP genetic studies and draw better conclusions. The most broadly studied polymorphism is TaqIB for which over 10,000 individuals have been genotyped and had HDL levels determined. When these studies are subjected to a meta-analysis, the B2B2 homozygotes are found to have higher HDL levels than B1B1 homozygotes (0.12 mmol/l, 95% CI = 0.11 0.13, P < 0.0001). A similar analysis of the I405V polymorphism yields 0.05 mmol/l higher HDL levels in 405VV homozygotes than in 405II homozygotes (95% CI = 0.03-0.07, P < 0.0001). The implications of these studies for cardiovascular disease will be addressed. PMID- 12639976 TI - Familial hypobetalipoproteinemia: a review. AB - We review the genetics and pathophysiology of familial hypobetalipoproteinemia (FHBL), a mildly symptomatic genetically heterogeneous autosomal trait. The minority of human FHBL is caused by truncation-specifying mutations of the APOB gene on chromosome 2. In seven families, linkage to chromosome 2 is absent, linkage is instead to chromosome 3 (3p21). In others, linkage is absent to both APOB and to 3p21. Apolipoprotein B-100 (apoB-100) levels are approximately 25% of normal, instead of the 50% expected based on the presence of one normal allele due to reduced rates of production. The presence of the truncating mutation seems to have a "dominant recessive" effect on apoB-100 secretion. Concentrations of apoB truncations in plasma differ by truncation but average at approximately 10% of normal levels. Lipoproteins bearing truncated forms of apoB are cleared more rapidly than apoB-100 particles. In contrast with apoB-100 particles cleared primarily in liver via the LDL receptor, most apoB truncation particles are cleared in renal proximal tubular cells via megalin. Since apoB defects cause a dysfunctional VLDL-triglyceride transport system, livers accumulate fat. Hepatic synthesis of fatty acids is reduced in compensation. Informational lacunae remain about genes affecting fat accumulation in liver, and the modulation of liver fat in the presence apoB truncation defects. PMID- 12639977 TI - Membrane dynamics and cell polarity: the role of sphingolipids. AB - In recent years, glycosphingolipids (GSLs) have attracted widespread attention due to the appreciation that this class of lipids has a major impact on biological life. Inhibition of the synthesis of glucosylceramide, which serves as a precursor for the generation of complex glycosphinglipids, is embryonic lethal. GSLs play a major role in growth and development. Metabolites of sphingolipids, such as ceramide, sphinganine, and sphingosine, may function as second messengers or regulators of signal transduction that affect events ranging from apoptosis to the (co)regulation of the cell cycle. In addition, GSLs can provide a molecular platform for clustering of signal transducers. The ability of sphingolipids, with or without cholesterol, to form microdomains or rafts is critical in sorting and membrane transport that underlies the biogenesis of polarized membrane domains. Here, a brief summary is presented of some recent developments in this field, with a particular emphasis on raft assembly and membrane transport in the establishment of membrane polarity. PMID- 12639978 TI - The role of medical school admissions committees in the decline of physician scientists. PMID- 12639979 TI - Specificity of a third kind: reactive oxygen and nitrogen intermediates in cell signaling. PMID- 12639980 TI - HIF hydroxylation and the mammalian oxygen-sensing pathway. PMID- 12639981 TI - Oxidative stress, cell cycle, and neurodegeneration. PMID- 12639982 TI - Can SOCS make arthritis better? PMID- 12639983 TI - Autonomic "myasthenia": the case for an autoimmune pathogenesis. PMID- 12639984 TI - Bone marrow: an extra-pancreatic hideout for the elusive pancreatic stem cell? PMID- 12639985 TI - The challenge of molecular medicine: complexity versus Occam's razor. PMID- 12639986 TI - Measles virus infection results in suppression of both innate and adaptive immune responses to secondary bacterial infection. AB - Among infectious agents, measles virus (MV) remains a scourge responsible for 1 million deaths per year and is a leading cause of childhood deaths in developing countries. Although MV infection itself is not commonly lethal, MV-induced suppression of the immune system results in a greatly increased susceptibility to opportunistic bacterial infections that are largely responsible for the morbidity and mortality associated with this disease. Despite its clinical importance, the underlying mechanisms of MV-induced immunosuppression remain unresolved. To begin to understand the basis of increased susceptibility to bacterial infections during MV infection, we inoculated transgenic mice expressing the MV receptor, CD46, with MV and Listeria monocytogenes. We found that MV-infected mice were more susceptible to infection with Listeria and that this corresponded with significantly decreased numbers of macrophages and neutrophils in the spleen and substantial defects in IFN-gamma production by CD4(+) T cells. The reduction in CD11b(+) macrophages and IFN-gamma-producing T cells was due to reduced proliferative expansion and not to enhanced apoptosis or to altered distribution of these cells between spleen, blood, and the lymphatic system. These results document that MV infection can suppress both innate and adaptive immune responses and lead to increased susceptibility to bacterial infection. PMID- 12639987 TI - Spontaneous circulation of myeloid-lymphoid-initiating cells and SCID repopulating cells in sickle cell crisis. AB - The only curative therapy for sickle cell disease (SCD) is allogeneic hematopoietic stem cell (HSC) transplantation. Gene therapy approaches for autologous HSC transplantation are being developed. Although earlier engraftment is seen when cells from GCSF-mobilized blood are transplanted than when bone marrow is transplanted, administration of GCSF to patients with SCD can cause significant morbidity. We tested whether primitive hematopoietic progenitors are spontaneously mobilized in the blood of patients with SCD during acute crisis (AC SCD patients). The frequency of myeloid-lymphoid-initiating cells (ML-ICs) and SCID-repopulating cells (SRCs) was significantly higher in blood from AC-SCD patients than in blood from patients with steady-state SCD or from normal donors. The presence of SRCs in peripheral blood was not associated with detection of long-term culture-initiating cells, consistent with the notion that SRCs are more primitive than long-term culture-initiating cells. As ML-ICs and SRCs were both detected in blood of AC-SCD patients only, these assays may both measure primitive progenitors. The frequency of ML-ICs also correlated with increases in stem cell factor, GCSF, and IL-8 levels in AC-SCD compared with steady-state SCD and normal-donor sera. Because significant numbers of ML-ICs and SRCs are mobilized in the blood without exogenous cytokine treatment during acute crisis of SCD, collection of peripheral blood progenitors during crisis may yield a source of autologous HSCs suitable for ex-vivo correction by gene therapy approaches and subsequent transplantation. PMID- 12639988 TI - Mechanisms of TNF-alpha- and RANKL-mediated osteoclastogenesis and bone resorption in psoriatic arthritis. AB - Psoriatic arthritis (PsA) is an inflammatory joint disease characterized by extensive bone resorption. The mechanisms underlying this matrix loss have not been elucidated. We report here that blood samples from PsA patients, particularly those with bone erosions visible on plain radiographs, exhibit a marked increase in osteoclast precursors (OCPs) compared with those from healthy controls. Moreover, PsA PBMCs readily formed osteoclasts in vitro without exogenous receptor activator of NF-kappaB ligand (RANKL) or MCSF. Both osteoprotegerin (OPG) and anti-TNF antibodies inhibited osteoclast formation. Additionally, cultured PsA PBMCs spontaneously secreted higher levels of TNF alpha than did healthy controls. In vivo, OCP frequency declined substantially in PsA patients following treatment with anti-TNF agents. Immunohistochemical analysis of subchondral bone and synovium revealed RANK-positive perivascular mononuclear cells and osteoclasts in PsA specimens. RANKL expression was dramatically upregulated in the synovial lining layer, while OPG immunostaining was restricted to the endothelium. These results suggest a model for understanding the pathogenesis of aggressive bone erosions in PsA. OCPs arise from TNF-alpha-activated PBMCs that migrate to the inflamed synovium and subchondral bone, where they are exposed to unopposed RANKL and TNF-alpha. This leads to osteoclastogenesis at the erosion front and in subchondral bone, resulting in a bidirectional assault on psoriatic bone. PMID- 12639989 TI - The role of the Grb2-p38 MAPK signaling pathway in cardiac hypertrophy and fibrosis. AB - Cardiac hypertrophy is a common response to pressure overload and is associated with increased mortality. Mechanical stress in the heart can result in the integrin-mediated activation of focal adhesion kinase and the subsequent recruitment of the Grb2 adapter molecule. Grb2, in turn, can activate MAPK cascades via an interaction with the Ras guanine nucleotide exchange factor SOS and with other signaling intermediates. We analyzed the role of the Grb2 adapter protein and p38 MAPK in cardiac hypertrophy. Mice with haploinsufficiency of the Grb2 gene (Grb2(+/-) mice) appear normal at birth but have defective T cell signaling. In response to pressure overload, cardiac p38 MAPK and JNK activation was inhibited and cardiac hypertrophy and fibrosis was blocked in Grb2(+/-) mice. Next, transgenic mice with cardiac-specific expression of dominant negative forms of p38alpha (DN-p38alpha) and p38beta (DN-p38beta) MAPK were examined. DN p38alpha and DN-p38beta mice developed cardiac hypertrophy but were resistant to cardiac fibrosis in response to pressure overload. These results establish that Grb2 action is essential for cardiac hypertrophy and fibrosis in response to pressure overload, and that different signaling pathways downstream of Grb2 regulate fibrosis, fetal gene induction, and cardiomyocyte growth. PMID- 12639991 TI - Acceleration of type 1 diabetes mellitus in proinsulin 2-deficient NOD mice. AB - Accumulating evidence favors a role for proinsulin as a key autoantigen in diabetes. In the mouse, two proinsulin isoforms coexist. Most studies point to proinsulin 2 as the major isoform recognized by T cells in the NOD mouse. We studied mice in which a null proinsulin 2 mutation was transferred from proinsulin 2-deficient 129 mice onto the NOD background along with 16 genetic markers (including I-A(g7) MHC molecule) associated with diabetes. Intercross mice from the fourth backcross generation showed that proinsulin 2(-/-) mice develop accelerated insulitis and diabetes. The high prevalence of anti-insulin autoantibodies in proinsulin 2(-/-) mice indicates that diabetes acceleration relates to altered recognition of proinsulin. The prevalence of anti-glutamic acid decarboxylase autoantibodies and of sialitis is not increased in proinsulin 2(-/-) mice. We give evidence that proinsulin 2 expression leads to silencing of T cells specific for an epitope shared by proinsulin 1 and proinsulin 2. In the human, alleles located in the VNTR region flanking the insulin gene control beta cell response to glucose and proinsulin expression in the thymus and are key determinants of diabetes susceptibility. Proinsulin 2(-/-) NOD mice provide a model to study the role of thymic expression of insulin in susceptibility to diabetes. PMID- 12639990 TI - In vivo derivation of glucose-competent pancreatic endocrine cells from bone marrow without evidence of cell fusion. AB - Bone marrow harbors cells that have the capacity to differentiate into cells of nonhematopoietic tissues of neuronal, endothelial, epithelial, and muscular phenotype. Here we demonstrate that bone marrow-derived cells populate pancreatic islets of Langerhans. Bone marrow cells from male mice that express, using a CRE LoxP system, an enhanced green fluorescent protein (EGFP) if the insulin gene is actively transcribed were transplanted into lethally irradiated recipient female mice. Four to six weeks after transplantation, recipient mice revealed Y chromosome and EGFP double-positive cells in their pancreatic islets. Neither bone marrow cells nor circulating peripheral blood nucleated cells of donor or recipient mice had any detectable EGFP. EGFP-positive cells purified from islets express insulin, glucose transporter 2 (GLUT2), and transcription factors typically found in pancreatic beta cells. Furthermore, in vitro these bone marrow derived cells exhibit - as do pancreatic beta cells - glucose-dependent and incretin-enhanced insulin secretion. These results indicate that bone marrow harbors cells that have the capacity to differentiate into functionally competent pancreatic endocrine beta cells and that represent a source for cell-based treatment of diabetes mellitus. The results generated with the CRE-LoxP system also suggest that in vivo cell fusion is an unlikely explanation for the "transdifferentiation" of bone marrow-derived cells into differentiated cell phenotypes. PMID- 12639992 TI - Rescue of cardiomyocyte dysfunction by phospholamban ablation does not prevent ventricular failure in genetic hypertrophy. AB - Cardiac hypertrophy, either compensated or decompensated, is associated with cardiomyocyte contractile dysfunction from depressed sarcoplasmic reticulum (SR) Ca(2+) cycling. Normalization of Ca(2+) cycling by ablation or inhibition of the SR inhibitor phospholamban (PLN) has prevented cardiac failure in experimental dilated cardiomyopathy and is a promising therapeutic approach for human heart failure. However, the potential benefits of restoring SR function on primary cardiac hypertrophy, a common antecedent of human heart failure, are unknown. We therefore tested the efficacy of PLN ablation to correct hypertrophy and contractile dysfunction in two well-characterized and highly relevant genetic mouse models of hypertrophy and cardiac failure, Galphaq overexpression and human familial hypertrophic cardiomyopathy mutant myosin binding protein C (MyBP C(MUT)) expression. In both models, PLN ablation normalized the characteristically prolonged cardiomyocyte Ca(2+) transients and enhanced unloaded fractional shortening with no change in SR Ca(2+) pump content. However, there was no parallel improvement in in vivo cardiac function or hypertrophy in either model. Likewise, the activation of JNK and calcineurin associated with Galphaq overexpression was not affected. Thus, PLN ablation normalized contractility in isolated myocytes, but failed to rescue the cardiomyopathic phenotype elicited by activation of the Galphaq pathway or MyBP-C mutations. PMID- 12639993 TI - Human phospholamban null results in lethal dilated cardiomyopathy revealing a critical difference between mouse and human. AB - In human disease and experimental animal models, depressed Ca(2+) handling in failing cardiomyocytes is widely attributed to impaired sarcoplasmic reticulum (SR) function. In mice, disruption of the PLN gene encoding phospholamban (PLN) or expression of dominant-negative PLN mutants enhances SR and cardiac function, but effects of PLN mutations in humans are unknown. Here, a T116G point mutation, substituting a termination codon for Leu-39 (L39stop), was identified in two families with hereditary heart failure. The heterozygous individuals exhibited hypertrophy without diminished contractile performance. Strikingly, both individuals homozygous for L39stop developed dilated cardiomyopathy and heart failure, requiring cardiac transplantation at ages 16 and 27. An over 50% reduction in PLN mRNA and no detectable PLN protein were noted in one explanted heart. The expression of recombinant PLN-L39stop in human embryonic kidney (HEK) 293 cells and adult rat cardiomyocytes showed no PLN inhibition of SR Ca(2+) ATPase and the virtual absence of stable PLN expression; where PLN was expressed, it was misrouted to the cytosol or plasma membrane. These findings describe a naturally-occurring loss-of-function human PLN mutation (PLN null). In contrast to reported benefits of PLN ablation in mouse heart failure, humans lacking PLN develop lethal dilated cardiomyopathy. PMID- 12639994 TI - DNA damage is a novel response to sublytic complement C5b-9-induced injury in podocytes. AB - In response to Ab-complement-mediated injury, podocytes can undergo lysis, apoptosis, or, when exposed to sublytic (<5% lysis) amounts of C5b-9, become activated. Following the insertion of sublytic quantities of C5b-9, there is an increase in signaling pathways and growth factor synthesis and release of proteases, oxidants, and other molecules. Despite an increase in DNA synthesis, however, sublytic C5b-9 is associated with a delay in G(2)/M phase progression in podocytes. Here we induced sublytic C5b-9 injury in vitro by exposing cultured rat podocytes or differentiated postmitotic mouse podocytes to Ab and a complement source; we also studied the passive Heymann nephritis model of experimental membranous nephropathy in rats. A major finding was that sublytic C5b-9-induced injury caused an increase in DNA damage in podocytes both in vitro and in vivo. This was associated with an increase in protein levels for p53, the CDK inhibitor p21, growth-arrest DNA damage-45 (GADD45), and the checkpoint kinases-1 and -2. Sublytic C5b-9 increased extracellular signal-regulated kinase 1 and -2 (ERK-1 and -2), and inhibiting ERK-1 and -2 reduced the increase in p21 and GADD45 and augmented the DNA damage response to sublytic C5b-9-induced injury. These results show that sublytic C5b-9 induces DNA damage in vitro and in vivo and may explain why podocyte proliferation is limited following immune mediated injury. PMID- 12639995 TI - Inactivation of hMLH1 and hMSH2 by promoter methylation in primary non-small cell lung tumors and matched sputum samples. AB - We performed a genetic and epigenetic study of the hMLH1 and hMSH2 mismatch repair genes in resected primary tumors from 77 non-small cell lung cancer (NSCLC) patients. The molecular alterations examined included the loss of mRNA and protein expression as well as promoter methylation, and the allelic imbalance of the chromosomal regions that harbor the genes. We found that 78% and 26% of patients showed at least one type of molecular alteration within the hMLH1 and hMSH2 genes, respectively. Promoter methylation of the hMLH1 gene was present in 55.8% of tumors, and was significantly associated with the reduction in mRNA and protein expression (P = 0.001). A 72% concordance of aberrant methylation in sputum samples with matched resected tumors was found. In addition, a 93% consistency between the promoter methylation and the mRNA expression of the hMSH2 gene was found in 14 female NSCLC patients. However, no correlation was found between the expression of hMLH1 and hMSH2 proteins and the allelic imbalance of five microsatellite markers closely linked to the genes. Our results suggest that hMLH1 is the major altered mismatch repair gene involved in NSCLC tumorigenesis, and that promoter methylation is the predominant mechanism in hMLH1 and hMSH2 deregulation. In addition, promoter methylation of the hMLH1 gene may be identified in sputum samples to serve as a potential diagnostic marker of NSCLC. PMID- 12639996 TI - Deficiency of cathepsin S reduces atherosclerosis in LDL receptor-deficient mice. AB - Human atherosclerotic lesions overexpress the lysosomal cysteine protease cathepsin S (Cat S), one of the most potent mammalian elastases known. In contrast, atheromata have low levels of the endogenous Cat S inhibitor cystatin C compared with normal arteries, suggesting involvement of this protease in atherogenesis. The present study tested this hypothesis directly by crossing Cat S-deficient (CatS(-/-)) mice with LDL receptor-deficient (LDLR(-/-)) mice that develop atherosclerosis on a high-cholesterol diet. Compared with LDLR(-/-) mice, double-knockout mice (CatS(-/-)LDLR(-/-)) developed significantly less atherosclerosis, as indicated by plaque size (plaque area and intimal thickening) and stage of development. These mice also had markedly reduced content of intimal macrophages, lipids, smooth muscle cells, collagen, CD4(+) T lymphocytes, and levels of IFN-gamma. CatS(-/-)LDLR(-/-) monocytes showed impaired subendothelial basement membrane transmigration, and aortas from CatS(-/-)LDLR(-/-) mice had preserved elastic laminae. These findings establish a pivotal role for Cat S in atherogenesis. PMID- 12639997 TI - Immunization with neuronal nicotinic acetylcholine receptor induces neurological autoimmune disease. AB - Neuronal nicotinic AChRs (nAChRs) are implicated in the pathogenesis of diverse neurological disorders and in the regulation of small-cell lung carcinoma growth. Twelve subunits have been identified in vertebrates, and mutations of one are recognized in a rare form of human epilepsy. Mice with genetically manipulated neuronal nAChR subunits exhibit behavioral or autonomic phenotypes. Here, we report the first model of an acquired neuronal nAChR disorder and evidence for its pertinence to paraneoplastic neurological autoimmunity. Rabbits immunized once with recombinant alpha3 subunit (residues 1-205) develop profound gastrointestinal hypomotility, dilated pupils with impaired light response, and grossly distended bladders. As in patients with idiopathic and paraneoplastic autoimmune autonomic neuropathy, the severity parallels serum levels of ganglionic nAChR autoantibody. Failure of neurotransmission through abdominal sympathetic ganglia, with retention of neuronal viability, confirms that the disorder is a postsynaptic channelopathy. In addition, we found ganglionic nAChR protein in small-cell carcinoma lines, identifying this cancer as a potential initiator of ganglionic nAChR autoimmunity. The data support our hypothesis that immune responses driven by distinct neuronal nAChR subtypes expressed in small cell carcinomas account for several lung cancer-related paraneoplastic disorders affecting cholinergic systems, including autoimmune autonomic neuropathy, seizures, dementia, and movement disorders. PMID- 12639998 TI - Suppressor of cytokine signaling-1 regulates acute inflammatory arthritis and T cell activation. AB - Suppressor of cytokine signaling-1 (SOCS-1) is a negative regulator of cytokine signaling. To investigate the role of SOCS-1 in regulating inflammatory and immune responses in disease, acute inflammatory arthritis was induced in mice lacking SOCS-1. Expression of SOCS-1 protein was detected within synovial granulomas and pannus tissue of WT mice by day 7 following induction of acute arthritis. The severity of synovial inflammation and joint destruction at the peak of disease was greater in the absence of SOCS-1, although disease resolution occurred normally. There was an increased percentage of myeloid cells infiltrating the synovium in mice lacking SOCS-1, and SOCS-1 promoter activity was present in synovial macrophages, lymphocytes, and fibroblasts, but not granulocytes. The T cell response in draining LNs was also dysregulated, as popliteal LNs from mice lacking SOCS-1 contained approximately fivefold more cells at the peak of acute arthritis. These cells were hyperproliferative on exposure to antigen in vitro, and purified splenic CD4(+) T cells from mice lacking SOCS-1 proliferated more strongly in response to stimulation with anti CD3. Reporter gene expression was detected in CD4(+) T cells bearing the activation markers CD25, CD44, and CD69. SOCS-1 is therefore expressed in hematopoietic and nonhematopoietic cell types in vivo and is an important regulator of acute inflammatory arthritis and of CD4(+) T cell activation. PMID- 12640001 TI - The cartography of breathing. PMID- 12640003 TI - Has the phoenix risen? PMID- 12640002 TI - C-terminal interaction of KCNQ2 and KCNQ3 K+ channels. AB - Coexpression of KCNQ2 and KCNQ3 channels results in a 10-fold increased current amplitude compared to that of KCNQ2 alone, suggesting the formation of heteromultimeric channels. There is no interaction of either channel with KCNQ1. We evaluated the C-terminus as a potential interaction domain by construction of chimeras with interchanged C-termini of KCNQ1, KCNQ2 and KCNQ3 and functional expression in Xenopus oocytes. The chimera of KCNQ1 with a KCNQ2 C-terminus (Q1ctQ2) showed an 8-fold increase in current amplitude, and Q1ctQ3 a 3-fold increase when coexpressed with KCNQ3 and KCNQ2, respectively, indicating that the C-terminus contains an interaction domain. To characterize this interacting region, we studied further chimeras of KCNQ1 containing different parts of the KCNQ3 C-terminus for interaction with KCNQ2. We also evaluated short sequences of the KCNQ2 C-terminus for a dominant-negative effect on Q1ctQ3. According to the results of these experiments, functional interaction of KCNQ2 and KCNQ3 requires a highly conserved region of about 80 amino acids, previously called the A domain, plus either 40 residues downstream of the A-domain (B-domain) or the proximal C-terminus between S6 and the A-domain. Furthermore, the chimeras Q1ctQ3 and Q2ctQ3 showed > 10-fold increased current amplitudes compared to KCNQ1 or KCNQ2 alone and a strong depolarizing shift of voltage-dependent activation. The proximal part of the KCNQ3 C-terminus was necessary to produce these effects. Our results indicate that specific parts of the C-terminus enable the interaction between KCNQ2 and KCNQ3 channels and that different parts of the KCNQ3 C-terminus are important for regulating current amplitude. PMID- 12640004 TI - The fetal dilemma: spare the brain and spoil the liver. PMID- 12640005 TI - Regulation of kainate receptors by protein kinase C and metabotropic glutamate receptors. AB - Kainate receptors have recently been shown to be involved in synaptic transmission, to regulate transmitter release and to mediate synaptic plasticity in different regions of the CNS. However, very little is known about endogenous mechanisms that can control native kainate receptor signalling. In this study we have found that GluR5-containing kainate receptor-mediated actions can be modulated by activation of protein kinase C (PKC) but not protein kinase A (PKA). However, both PKA and PKC directly phosphorylate the GluR5 subunit of kainate receptors. Metabotropic glutamate (mGlu) receptors are well known to be involved in synaptic transmission, regulation of transmitter release and synaptic plasticity in a variety of brain regions. We now demonstrate that kainate receptor signalling is enhanced by activation of group I mGlu receptors, in a PKC dependent manner. These data demonstrate for the first time that kainate receptor function can be modulated by activation of metabotropic glutamate receptors and have implications for understanding mechanisms of synaptic transmission, plasticity and disorders such as epilepsy. PMID- 12640006 TI - Reflex sympathetic activation during static exercise is severely impaired in patients with myophosphorylase deficiency. AB - During static exercise, metabolites accumulate in the muscle interstitium where they stimulate chemosensitive afferent nerves that reflexly increase efferent muscle sympathetic nerve activity (MSNA) and blood pressure. In experimental animals, lactic acid potently stimulates the muscle metaboreflex, but its role in humans is more controversial. To determine if lactic acid is a critical mediator of metaboreflex activation in humans, we performed microelectrode recordings of MSNA in eight patients with myophosphorylase deficiency (McArdle's disease) who cannot metabolize intramuscular glycogen and do not generate lactic acid in exercising muscles. Each patient was matched with three healthy control subjects to maximize statistical power. In controls, 2 min of static handgrip performed at 33 % or 45 % of maximal voluntary contraction (MVC) produced intensity-dependent increases in MSNA (171 +/- 22 % and 379 +/- 95 %, respectively). In the patients, MSNA responses to static handgrip were markedly attenuated (33 +/- 14 % at 33 % MVC; 32 +/- 19 % at 45 % MVC; P < 0.05 vs. controls). Likewise, when static handgrip (30 % MVC) was performed to fatigue, MSNA increased by 366 +/- 73 % in controls but only by 51 +/- 14 % in patients (P < 0.05). Pressor responses to static handgrip were also attenuated in patients compared to controls, whereas heart rate responses were identical. In contrast to exercise, the MSNA responses to other reflex stimuli (the cold pressor test or Valsalva's manoeuvre) were similar in patients and controls. Together these data indicate that appropriate activation of glycogenolytic pathways is obligatory for normal metaboreflex mediated sympathoexcitation during static exercise in humans. PMID- 12640007 TI - The action of BDNF on GABA(A) currents changes from potentiating to suppressing during maturation of rat hippocampal CA1 pyramidal neurons. AB - During the development of the hippocampus, the action of GABA shifts from depolarizing to hyperpolarizing, and brain-derived neurotrophic factor (BDNF) has important roles in GABAergic transmission. We demonstrate that BDNF (20 ng ml-1) rapidly and reversibly potentiates postsynaptic GABAA receptor-mediated currents (by 80.5 +/- 14.3 %, n = 10) in hippocampal CA1 pyramidal neurons isolated from postnatal day (P)6 rats, using nystatin-perforated patch-clamp recordings. This potentiation is caused by an elevation of intracellular Ca2+ that occurs in response to the activation of Trk B receptor tyrosine kinase and phospholipase C gamma. The modulation of the GABAA responses by BDNF in hippocampal CA1 pyramidal neurons isolated from P10 rats was more diverse (from potentiating to inhibitory), and at P14, BDNF induced a long-lasting inhibition. In addition, Ca2+/calmodulin-dependent protein kinase 2 plays important roles in the potentiating, but not in the inhibitory effect, of BDNF on the GABAA responses. These results suggest that changes in the intracellular signalling pathway could contribute to the developmental shift of the actions of BDNF on inhibitory systems. PMID- 12640008 TI - Voltage-dependent gating and block by internal spermine of the murine inwardly rectifying K+ channel, Kir2.1. AB - The mechanism of inward rectification was investigated by recording single channel currents through an inwardly rectifying K+ channel (Kir2.1). cDNA encoding a wild-type (WT) channel, a mutant replacing Asp 172 with Asn (D172N), and a tandem tetramer WT-(D172N)2-WT, was transfected into COS-1 cells using the liposome method, and after 48-72 h single-channel currents were recorded in the inside-out configuration at 150 mM internal and external K+. Steady-state open probability of outward currents decreased with larger depolarizations. The activation curve was fitted with a single Boltzmann equation. The voltages of half-activation in the absence of spermine were +35.9 mV (WT), +55.0 mV (WT (D172N)2-WT) and +76.7 mV (D172N). Open-time and zero-current-time histograms were constructed. The open-time histogram was fitted with a single exponential function. Two exponential functions were necessary to fit the closed-time histogram. In each channel, internal spermine at a concentration of 1-100 nM reduced the open time of the outward currents in a concentration-dependent manner and produced one blocked state without affecting the inward currents, suggesting that spermine acts as an open channel blocker. The normalized steady-state open probability-spermine concentration curve was fitted by saturation kinetics with a Hill coefficient of 1. On the assumption of the linear sequential state model, the unblock and blocking rates were estimated in each channel. Unblock rates depended on the number of D172N mutant subunits, but blocking rates did not. The results suggest that closing gates work independently of the spermine block and D172 is involved in both intrinsic gating and the spermine block. PMID- 12640009 TI - Defining ventral medullary respiratory compartments with a glutamate receptor agonist in the rat. AB - The regional organization of the ventral respiratory group (VRG) was examined with respect to generation of respiratory rhythm (breathing frequency) versus control of the respiratory motor pattern on individual nerves. In urethane anaesthetized, neuromuscularly blocked and vagotomized Sprague-Dawley rats, arterial blood pressure (ABP) and respiratory motor outputs (phrenic, pharyngeal branch of the vagus, or superior laryngeal nerves) were recorded. The VRG organization was mapped systematically using injections of the excitatory amino acid DL-homocysteic acid (DLH; 5-20 mM, 2-6 nl) from single- or double-barrel pipettes at 100-200 microm intervals between the facial nucleus and the calamus scriptorius. Recording of respiratory neurons through the injection pipette ensured that the pipette was located within the VRG. At the end of each experiment, the injection pipette was used to make an electrical lesion, thereby marking the electrode position for subsequent histological reconstruction of injection sites. Four rostrocaudal regions were identified: (1) a rostral bradypnoea area, at the level of the Botzinger complex, in which respiratory rhythm slowed and ABP increased, (2) a tachypnoea/dysrhythmia area, at the level of the preBotzinger complex, in which breathing rate either increased or became irregular, with little or no change in ABP, (3) a caudal bradypnoea area at the level of the anterior part of the rostral VRG in which ABP decreased and (4) a caudal 'no effect' region in the posterior part of the rostral VRG. The peak amplitude of phrenic nerve activity decreased with injections into all three rostral regions. Changes in respiratory rhythm were associated with opposite changes in inspiratory (TI) and expiratory (TE) durations after injections into either the Botzinger complex or anterior rostral VRG, while both TI and TE decreased after injections into the preBotzinger complex. Effects on selected cranial nerves were similar to those on the phrenic nerve except that tonic activity was elicited on the superior larygneal nerve ipsilateral to injections in the Botzinger complex and on the pharyngeal branch of the vagus ipsilateral to injections in the preBotzinger complex. These data reinforce the subdivision of the VRG into functionally distinct compartments and suggest that a further subdivision of the rostral VRG is warranted. They also suggest that region specific influences, especially on the pattern of cranial motor discharge, can be used to assist the identification of recording sites within the VRG. However, the postulated clear functional separation of rhythm- versus pattern-generating regions was not supported. PMID- 12640010 TI - Prostaglandins but not nitric oxide protect renal medullary perfusion in anaesthetised rats receiving angiotensin II. AB - Angiotensin II (Ang II) fails to constrict renal medullary vasculature, possibly due to the counteraction of local vasodilators, such as prostaglandins or nitric oxide (NO). The effects of exogenous Ang II on intrarenal circulation were determined in anaesthetised rats that were untreated or pretreated with indomethacin (Indo) or L-NAME. The total renal blood flow (RBF), representing cortical perfusion, and outer and inner medullary blood flow (OMBF and IMBF) were measured. In untreated rats, Ang II decreased RBF in a dose dependent manner. Intravenous administration of 30 ng kg-1 min-1 Ang II decreased RBF by 38 % and OMBF by 9 % (both significant); IMBF was unaffected. Indo (5 mg kg-1 I.V.) significantly and similarly decreased OMBF and IMBF without affecting RBF. Ang II decreased IMBF by 27 % in Indo-pretreated rats, but caused no change in rats without pretreatment. The decreases in OMBF and RBF were comparable with or without Indo pretreatment. Inhibition of NO synthesis with L-NAME (0.6 mg kg-1 I.V.) significantly decreased RBF, OMBF and IMBF. Ang II infusion into L-NAME pretreated rats induced a further significant decrease in RBF and OMBF without changing IMBF. We conclude that within the inner medulla, but not the outer medulla or cortex, prostaglandins effectively counteract the vasopressor effect of circulating Ang II. PMID- 12640011 TI - Brain-derived neurotrophic factor attenuates mouse cerebellar granule cell GABA(A) receptor-mediated responses via postsynaptic mechanisms. AB - In addition to exerting long-term neurotrophic influences on developmental process such as neuronal survival and neuritic outgrowth, brain-derived neurotrophic factor (BDNF) has been reported to modulate synaptic transmission in the short-term. Considerable evidence indicates that BDNF acutely modulates NMDA receptor-mediated synaptic activity. However, whether BDNF modulates inhibitory synaptic transmission remains to be firmly established. In the present study, we examined the effect of acute BDNF exposure on GABA-evoked whole-cell responses as well as GABAergic synaptic activity in cultured mouse cerebellar granule cells. GABA-evoked responses were reduced by 39.5 +/- 4.7 % upon acute and focal application of BDNF (100 ng ml-1). The reduction of the GABA response recovered only partially even minutes after removal of BDNF. TrkB-IgG and K252a, but not K252b, prevented the BDNF-induced attenuation of the GABA response. BDNF exposure shifted the cumulative peak amplitude distribution leftward for both spontaneous IPSCs (sIPSCs) and miniature IPSCs (mIPSCs) without affecting the rise time and decay time constants. Acute exposure to BDNF also resulted in internalization of GABAA receptors in cultured cerebellar granule cells, as reflected by diminished immunostaining with an antibody against the GABAA receptor beta2/3 subunit. Although the BDNF-induced GABAA receptor internalization was sensitive to K252a, it did not become manifest until 5 min after exposure to BDNF. Therefore, receptor internalization alone cannot account for the prompt BDNF-induced attenuation of GABA-mediated activity. We conclude that BDNF modulates GABAA receptor-mediated activity through TrkB receptor signalling that triggers a kinase-dependent short latency effect and a delayed longer latency effect hallmarked by receptor internalization. PMID- 12640012 TI - Axonal propagation: does the spike stop here? PMID- 12640013 TI - PKA-mediated inhibition of a novel K+ channel underlies the slow after hyperpolarization in enteric AH neurons. AB - Postspike after-hyperpolarizations (AHPs) control the excitability of neurons and are important in shaping firing patterns. The duration of some of these events extends to tens of seconds and they can render neurons inexcitable for much of their time course. While consensus is strong that the medium duration (< 1 s AHPs are mediated by the opening of small conductance Ca2+-activated K+ channels, the K+ channels mediating slow AHPs (> 5 s in a subset of enteric (AH) neurons) have an intermediate unit conductance (IKCa). Using whole-cell and excised-patch recording, we have demonstrated that the cAMP-protein kinase A (PKA) pathway regulates the activity of these channels. In whole-cell mode, forskolin (0.003-1 microM) inhibited the current underlying the slow AHP (IsAHP) by 90 %, and this was partially sensitive to inhibition of PKA with internal Rp-cAMPS (500 microM). Rp-cAMPS alone increased the current following break-in and caused a 20 mV hyperpolarization, suggesting that PKA maintains slow AHP channels in the closed state. Internal perfusion of the inhibitory peptide PKI5-24 slightly increased the IsAHP and opposed the inhibitory action of forskolin. Internal perfusion of the catalytic subunit of PKA (PKAcat) suppressed the IsAHP by 50 % without affecting membrane potential or action potential configuration. In inside-out patches containing IKCa-like channels, PKAcat decreased the open probability of IKCa-like channels while alkaline phosphatase activated them. These results suggest that the IKCa-like channels that underlie the slow AHP in myenteric AH neurons are subject to inhibition by PKA-dependent phosphorylation and that PKA plays an integral role in their gating. PMID- 12640014 TI - Regulation by voltage and adenine nucleotides of a Ca2+-activated cation channel from hamster vomeronasal sensory neurons. AB - Bipolar sensory neurons within the vomeronasal organ (VNO) are thought to mediate the detection of pheromones in vertebrates. In the mouse, VNO neurons respond to pheromones with a rise in intracellular Ca2+ that accompanies a depolarization of the cell. Transduction of the pheromone appears to occur through the activation of a phosphatidylinositol signalling pathway, but the ion channels that respond to this signalling pathway have not been identified. In this report patch-clamp recording from hamster vomeronasal sensory neurons was used to identify second messenger-gated channels that might play a role in transduction. The results demonstrate that VNO neurons show abundant expression of a Ca2+-activated non selective (CaNS) cation channel. The CaNS channel does not discriminate between Na+ and K+ and has a slope conductance of 22 pS. Half-activation of the channel occurs at a Ca2+ concentration of 0.5 mM (at -80 mV). The probability of opening (Po) of the channel is further augmented at positive potentials, and shows an e fold voltage dependence per 37 mV. The channel exhibits rapid rundown following patch excision with Po decreasing from near 1.0 to near 0. The adenine nucleotides ATP and cAMP block the channel with an apparent affinity of 3 and 42 microM, respectively (-80 mV). Both the activation of the channel by Ca2+ and the block of the channel by adenine nucleotides show a mild voltage dependence, which can be accounted for by the voltage dependence of Po. The properties of this channel make it a candidate to either directly mediate vomeronasal sensory transduction, or to amplify the primary sensory response. PMID- 12640015 TI - Intraterminal Ca2+ concentration and asynchronous transmitter release at single GABAergic boutons in rat collicular cultures. AB - Neurotransmitter release in response to a single action potential has a precise time course. A significant fraction of the releasable vesicles is exocytosed synchronously, within a few milliseconds after the arrival of an action potential. If repeatedly activated, stimulus-locked phasic synchronous release declines, but synaptic transmission can be maintained through tonic asynchronous transmitter release. The desynchronisation of release during repetitive activation is generally attributed to a build-up of intraterminal Ca2+ concentration. However, the precise relationship between presynaptic Ca2+ level and asynchronous release rate at small central synapses has remained unclear. Here we characterise this relationship for single GABAergic terminals in rat collicular cultures. In the presence of tetrodotoxin, inhibitory postsynaptic currents (IPSCs) and presynaptic Ca2+ transients were recorded in response to direct presynaptic depolarisation of individual boutons. Repetitive stimulation indeed resulted in a shift from phasic to asynchronous neurotransmitter release. A clear dominance of the asynchronous release mode was observed after 10 pulses. The steady-state asynchronous release rate showed a third-power dependency on the presynaptic Ca2+ concentration, which is similar to that of evoked release. The Ca2+ sensor for asynchronous release exhibited a high affinity for Ca2+ and was far from saturation. These properties of the Ca2+ sensor should make the asynchronous release very sensitive to any modification of presynaptic Ca2+ concentration, including those resulting from changes in presynaptic activity patterns. Thus, asynchronous release represents a powerful but delicately regulated mechanism that ensures the maintenance of appropriate inhibition when the readily releasable pool of vesicles is depleted. PMID- 12640016 TI - Developmental changes in contractility and sarcomeric proteins from the early embryonic to the adult stage in the mouse heart. AB - Developmental changes in force-generating capacity and Ca2+ sensitivity of contraction in murine hearts were correlated with changes in myosin heavy chain (MHC) and troponin (Tn) isoform expression, using Triton-skinned fibres. The maximum Ca2+-activated isometric force normalized to the cross-sectional area (FCSA) increased mainly during embryogenesis and continued to increase at a slower rate until adulthood. During prenatal development, FCSA increased about 5 fold from embryonic day (E)10.5 to E19.5, while the amount of MHC normalized to the amount of total protein remained constant (from E13.5 to E19.5). This suggests that the development of structural organization of the myofilaments during the embryonic and the fetal period may play an important role for the improvement of force generation. There was an overall decrease of 0.5 pCa units in the Ca2+ sensitivity of force generation from E13.5 to the adult, of which the main decrease (0.3 pCa units) occurred within a short time interval, between E19.5 and 7 days after birth (7 days pn). Densitometric analysis of SDS-PAGE and Western blots revealed that the major switches between troponin T (TnT) isoforms occur before E16.5, whereas the transition points of slow skeletal troponin I (ssTnI) to cardiac TnI (cTnI) and of beta-MHC to alpha-MHC both occur around birth, in temporal correlation with the main decrease in Ca2+ sensitivity. To test whether the changes in Ca2+ sensitivity are solely based on Tn, the native Tn complex was replaced in fibres from E19.5 and adult hearts with fast skeletal Tn complex (fsTn) purified from rabbit skeletal muscle. The difference in pre replacement values of pCa50 (-log([Ca2+] M-1)) required for half-maximum force development) between E19.5 (6.05 +/- 0.01) and adult fibres (5.64 +/- 0.04) was fully abolished after replacement with the exogenous skeletal Tn complex (pCa50 = 6.12 +/- 0.05 for both stages). This suggests that the major developmental changes in Ca2+ sensitivity of skinned murine myocardium originate primarily from the switch of ssTnI to cTnI. PMID- 12640017 TI - A novel O2-sensing mechanism in rat glossopharyngeal neurones mediated by a halothane-inhibitable background K+ conductance. AB - Modulation of K+ channels by hypoxia is a common O2-sensing mechanism in specialised cells. More recently, acid-sensitive TASK-like background K+ channels, which play a key role in setting the resting membrane potential, have been implicated in O2-sensing in certain cell types. Here, we report a novel O2 sensitivity mediated by a weakly pH-sensitive background K+ conductance in nitric oxide synthase (NOS)-positive neurones of the glossopharyngeal nerve (GPN). This conductance was insensitive to 30 mM TEA, 5 mM 4-aminopyridine (4-AP) and 200 microM Cd2+, but was reversibly inhibited by hypoxia (O2 tension (PO2) = 15 mmHg), 2-5 mM halothane, 10 mM barium and 1 mM quinidine. Notably, the presence of halothane occluded the inhibitory effect of hypoxia. Under current clamp, these agents depolarised GPN neurones. In contrast, arachidonic acid (5-10 microM) caused membrane hyperpolarisation and potentiation of the background K+ current. This pharmacological profile suggests the O2-sensitive conductance in GPN neurones is mediated by a class of background K+ channels different from the TASK family; it appears more closely related to the THIK (tandem pore domain halothane-inhibited K+) subfamily, or may represent a new member of the background K+ family. Since GPN neurones are thought to provide NO-mediated efferent inhibition of the carotid body (CB), these channels may contribute to the regulation of breathing during hypoxia via negative feedback control of CB function, as well as to the inhibitory effect of volatile anaesthetics (e.g. halothane) on respiration. PMID- 12640018 TI - Developmental regulation of hepatic and renal gluconeogenic enzymes by thyroid hormones in fetal sheep during late gestation. AB - Tissue glucose-6-phosphatase (G6P) and phosphoenolpyruvate carboxykinase (PEPCK) activities were investigated in sheep fetuses after experimental manipulation of thyroid hormone status. Increments in hepatic and renal G6P and PEPCK activities seen between 127-130 and 140-145 days of gestation (term, 145 +/- 2 days) were abolished when the normal prepartum rise in plasma triiodothyronine (T3), but not cortisol, was prevented by fetal thyroidectomy (TX). At 127-130 days, hepatic and renal G6P, and renal PEPCK, activities were similar in intact and TX fetuses; however, hepatic PEPCK was increased by TX. At 140-145 days, tissue G6P and PEPCK activities in TX fetuses were lower than in intact fetuses. In immature fetuses infused with cortisol (2-3 mg (kg body wt)-1 day-1) for five days, hepatic and renal enzyme activities were increased to those seen in mature fetuses near term. After five days of T3 infusion (8-12 microg (kg body wt)-1 day-1), G6P and PEPCK activities in the liver and kidney were greater than in saline-infused fetuses, but only renal G6P and PEPCK increased to the level seen close to term. Therefore, in fetal sheep, thyroid hormones are important for the prepartum rises in G6P and PEPCK activities in the liver and kidney and may mediate, in part, the maturational effects of cortisol. PMID- 12640019 TI - Compromised neuroimmune status in rats with experimental colitis. AB - In colitis, chronic and recurrent inflammation is associated with a breakdown in host defence mechanisms that leads to local and systemic infection. Whether this is due to a compromised neuroimmune response has not been studied. Our aim was to determine if colitis altered the host neuroimmune response as reflected in either body temperature rhythm or the febrile responses to lipopolysaccharide (LPS). Body temperature was monitored by telemetry from conscious, unrestrained male rats treated with trinitrobenzene sulphonic acid or saline. Twenty-six days after initial induction, colitis was reactivated. Animals were given LPS (50 microg kg 1 Escherichia coli LPS) during colitis and after reactivation. At the peak of colitis, treated rats showed a disruption of circadian body temperature rhythm, manifested as day-time fever followed by night-time hypothermia. In response to LPS, controls displayed a characteristic fever, whereas treated animals had a significantly reduced fever and low plasma levels of interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-alpha). During reactivation of colitis, treated animals did not mount a fever or exhibit increased plasma levels of IL-6 and TNF alpha after LPS. We conclude that experimental colitis is associated with a compromised neuroimmune status. PMID- 12640020 TI - Presence of (phospho)creatine in developing and adult skeletal muscle of mice without mitochondrial and cytosolic muscle creatine kinase isoforms. AB - We assessed the relationship between phosphocreatine (PCr) and creatine (Cr) content and creatine kinase (CK) activity in skeletal muscle of mice. The PCr and total Cr (tCr) concentrations, as well as CK activity, in hindlimb muscles of mice, with or without the cytosolic and mitochondrial isoforms of muscle creatine kinase (wild-type or CK--/-- mice), were determined by in vivo magnetic resonance (MR) spectroscopy and by biochemical means during postnatal growth and adulthood. In wild-type muscle the [tCr], PCr/ATP ratio and CK activity increased rapidly in the first 4-7 weeks. Remarkably, CK--/-- mice showed a similar increase in the PCr/ATP ratio during the first month in the presence of only minor brain-type BB CK activity. Uptake of Cr in muscle was seemingly unrelated to CK activity as tCr increased in the same way in the muscles of both mouse types. At older ages the PCr/ATP ratio decreased in CK--/-- muscles, in contrast to wild-type where it still slowly increased, whereas [tCr] was similar for muscle of both mouse types. Using a new in vivo MR approach with application of [4-13C]Cr, a lower PCr/tCr ratio was also observed in CK--/-- muscle. From these data it follows that in vivo global ATP levels at rest are similar in the presence or absence of CK. Although Cr could still be converted to PCr in mature CK--/-- muscle, the immediate availability of PCr decreased, and PCr became partly inconvertible at older ages. Apparently, catalysis of the CK reaction by BB-CK, although significant in muscles of newborn mice, gradually declines to very low levels in adulthood. Part or all of this BB-CK may arise from satellite cells fusing with myotubes, a process that is most active during the first months of life. Finally, our observation that the MR and chemical assessment of muscle [tCr] and PCr/tCr ratio were similar for all mice does not support the existence of a significant MR-invisible or immobile pool of Cr, with a role for CK in this phenomenon. PMID- 12640023 TI - Pressure-volume behaviour of the rat upper airway: effects of tongue muscle activation. AB - Our hypothesis was that the simultaneous activation of tongue protrudor and retractor muscles (co-activation) would constrict and stiffen the pharyngeal airway more than the independent activation of tongue protrudor muscles. Upper airway stiffness was determined by injecting known volumes of air into the sealed pharyngeal airway of the anaesthetized rat while measuring nasal pressure under control (no-stimulus) and stimulus conditions (volume paired with hypoglossal (XII) nerve stimulation). Stimulation of the whole XII nerves (co-activation) or the medial XII branches (protrudor activation) effected similar increases in total pharyngeal airway stiffness. Importantly, co-activation produced volume compression (airway narrowing) at large airway volumes (P < 0.05), but had no effect on airway dimension at low airway volumes. In comparison, protrudor activation resulted in significant volume expansion (airway dilatation) at low airway volumes and airway narrowing at high airway volumes (P < 0.05). In conclusion, both co-activation and independent protrudor muscle activation increase airway stiffness. However, their effects on airway size are complex and depend on the condition of the airway at the time of activation. PMID- 12640021 TI - Acute interleukin-6 administration does not impair muscle glucose uptake or whole body glucose disposal in healthy humans. AB - The cytokine interleukin (IL)-6 has recently been linked with type 2 diabetes mellitus and has been suggested to affect glucose metabolism. To determine whether acute IL-6 administration affects whole-body glucose kinetics or muscle glucose uptake, 18 healthy young men were assigned to one of three groups receiving a high dose of recombinant human IL-6 (HiIL-6; n = 6), a low dose of IL 6 (LoIL-6; n = 6) or saline (Con; n = 6) infused into one femoral artery for 3 h. The stable isotope [6,6-2H2] glucose was infused into a forearm vein throughout the 3 h infusion period and for a further 3 h after the cessation of infusion (recovery) to determine endogenous glucose production and whole-body glucose disposal. Infusion with HiIL-6 and LoIL-6 resulted in a marked (P < 0.05) increase in systemic IL-6 concentration throughout the 3 h of infusion (mean arterial plasma [IL-6]s of 319 and 143 pg ml-1 for HiIL-6 and LoIL-6, respectively), followed by a rapid decline (P < 0.05) during the recovery period. Subjects experienced clinical symptoms such as shivering and discomfort during HiIL-6 administration, but were asymptomatic during LoIL-6 administration. In addition, only HiIL-6 elevated (P < 0.05) plasma adrenaline (epinephrine). IL-6 infusion, irrespective of dose, did not result in any changes to endogenous glucose production, whole-body glucose disposal or leg- glucose uptake. These data demonstrate that acute IL-6 administration does not impair whole-body glucose disposal, net leg-glucose uptake, or increase endogenous glucose production at rest in healthy young humans. PMID- 12640022 TI - Single-axon action potentials in the rat hippocampal cortex. AB - Whether all action potentials propagate faithfully throughout axon arbors in the mammalian CNS has long been debated, and remains an important issue because many synapses occur far from the soma along extremely thin, unmyelinated, varicosity laden branches of axon arbors. We detected unitary action potentials along individual axon branches of adult hippocampal CA3 pyramidal cells using extracellular electrodes, and analysed their conduction across long distances (mean, 2.1 mm) at 22 and 37 degrees C. Axons nearly always transmitted low frequency impulses. At higher frequencies, most axons also transmitted impulses with striking fidelity. However, at paired-pulse frequencies in the hundreds of kilohertz range, axons exhibited variability: refractory periods ranged from 2.5 to 10 ms at 37 degrees C and from 5 to 40 ms at 22 degrees C. Although the basis for the refractory period variability could not be determined, these limits overlap with CA3 spike frequencies observed in vivo, raising the possibility that some axonal branches act as filters for the higher-order spikes in bursts, in contrast to the observed first-spike reliability. These results extend the observations of propagation reliability to a much longer distance and higher frequency domain than previously reported, and suggest a high safety factor for action potential propagation along thin, varicose axons. PMID- 12640024 TI - Antibiotics acting on the translational machinery. PMID- 12640025 TI - Tensegrity II. How structural networks influence cellular information processing networks. AB - The major challenge in biology today is biocomplexity: the need to explain how cell and tissue behaviors emerge from collective interactions within complex molecular networks. Part I of this two-part article, described a mechanical model of cell structure based on tensegrity architecture that explains how the mechanical behavior of the cell emerges from physical interactions among the different molecular filament systems that form the cytoskeleton. Recent work shows that the cytoskeleton also orients much of the cell's metabolic and signal transduction machinery and that mechanical distortion of cells and the cytoskeleton through cell surface integrin receptors can profoundly affect cell behavior. In particular, gradual variations in this single physical control parameter (cell shape distortion) can switch cells between distinct gene programs (e.g. growth, differentiation and apoptosis), and this process can be viewed as a biological phase transition. Part II of this article covers how combined use of tensegrity and solid-state mechanochemistry by cells may mediate mechanotransduction and facilitate integration of chemical and physical signals that are responsible for control of cell behavior. In addition, it examines how cell structural networks affect gene and protein signaling networks to produce characteristic phenotypes and cell fate transitions during tissue development. PMID- 12640026 TI - Focal adhesion kinase: the first ten years. AB - The protein tyrosine kinase focal adhesion kinase (FAK) plays a prominent role in integrin signaling. FAK activation, demonstrated by an increase in phosphorylation of Tyr397 as well as other sites in the protein, is best understood in the context of the engagement of integrins at the cell surface. Activation of FAK results in recruitment of a number of SH2-domain- and SH3 domain-containing proteins, which mediate signaling to several downstream pathways. FAK-dependent activation of these pathways has been implicated in a diverse array of cellular processes, including cell migration, growth factor signaling, cell cycle progression and cell survival. PMID- 12640027 TI - Observation of keratin particles showing fast bidirectional movement colocalized with microtubules. AB - Keratin intermediate filament networks were observed in living cultured epithelial cells using the incorporation of fluorescently tagged keratin from a transfected enhanced green fluorescent protein (EGFP) construct. In steady-state conditions EGFP-keratin exists not only as readily detectable intermediate filaments, but also as small particles, of which there are two types: a less mobile population (slow or static S particles) and a highly dynamic one (fast or F particles). The dynamic F particles move around the cell very fast and in a non random way. Their movement is composed of a series of steps, giving an overall characteristic zig-zag trajectory. The keratin particles are found all over the cell and their movement is aligned with microtubules; treatment of cells with nocodazole has an inhibitory effect on keratin particle movement, suggesting the involvement of microtubule motor proteins. Double-transfection experiments to visualize tubulin and keratin together suggest that the movement of keratin particles can be bidirectional, as particles are seen moving both towards and away from the centrosome area. Using field emission scanning and transmission electron microscopy combined with immunogold labelling, we also detected particulate keratin structures in untransfected epithelial cells, suggesting that keratin particles may be a natural component of keratin filament dynamics in living cells. PMID- 12640028 TI - Chronic acid exposure leads to activation of the cdx2 intestinal homeobox gene in a long-term culture of mouse esophageal keratinocytes. AB - To explore mechanisms whereby Malpighian keratinocytes can transdifferentiate into an intestinal-like epithelium, as observed in the early steps of Barrett's esophagus (BE) development, long-standing cultures of esophageal keratinocytes derived from normal mouse esophageal explants were developed. These cells were able to form multilayers and to differentiate on filter support by the formation of differentiated layers of basal cells (cytokeratine 14 positive) on which secondary suprabasal cell layers (cytokeratine 4 positive) spontaneously developed. Thus, these cultured cells, referred to as P3E6, reproduced, at least in part, the proliferation and stratification pattern existing in the normal esophagus. Because chronic exposure to acid pH is known to be a critical factor for BE development, culture medium at pH 3.5 was added into the apical chamber of cell cultures. This led to a decrease in the overall number of cells but it did not affect cell proliferation. Furthermore, external acid environment triggered expression of the GFP reporter gene fused downstream of the cdx2 intestinal homeogene regulatory sequences in P3E6 transfected cells. Expression of the endogenous CDX2 protein, detected by western blot and immunocytochemical analysis, correlated with promoter activation. These findings demonstrate that chronic exposure of esophageal keratinocytes to acid pH induces transcription of cdx2, an intestinal specific homeobox gene known to play a critical role in the differentiation and maintenance of intestinal epithelial functions. The results suggest that chronic acid exposure can modify the fate of P3E6 esophageal keratinocytes towards an intestinal program. This can be a key step in the development of intestinal metaplasia often observed in esophagus-cardia junction. PMID- 12640029 TI - Two phases of signalling between mitochondria during apoptosis leading to early depolarisation and delayed cytochrome c release. AB - We investigated the mode of signalling between mitochondria during apoptosis by monitoring the behaviour of non-irradiated mitochondria following microscopic photosensitisation of half the mitochondria in single human osteosarcoma cells loaded with CMXRos. Following partial irradiation of cells, non-irradiated mitochondria underwent a rapid depolarisation (within 10 minutes). The depolarisation was not inhibited by the caspase inhibitor zVAD-fmk but was suppressed by the intracellular Ca(2+) chelator BAPTA and overexpression of Bcl 2. Significantly, such depolarisation occurred even after prior conversion of extended filamentous mitochondria into individual punctate structures, indicating that lumenal continuity is not required for communication between the irradiated and non-irradiated mitochondria. Partial irradiation of cells expressing cytochrome c-GFP revealed cytochrome c-GFP release from non-irradiated mitochondria at a delayed but unpredictable time interval (between 30 minutes and more than 2.5 hours) following irradiation, which was unaffected by zVAD-fmk. Once activated, cytochrome c-GFP release occurred within a 10 minute period. Immunocytochemistry failed to reveal the recruitment of Bax to non-irradiated mitochondria, which suggests that Bax does not mediate the release of cytochrome c from mitochondria. We conclude that signals (mediated by Ca(2+)) emanating from irradiated mitochondria are processed by their non-irradiated counterparts and comprise two temporally distinct phases, both independent of caspase-mediated amplification, which generate an initial rapid depolarisation and subsequent delayed release of cytochrome c. PMID- 12640030 TI - Chlamydomonas DIP13 and human NA14: a new class of proteins associated with microtubule structures is involved in cell division. AB - We have cloned and characterized a single copy C. reinhardtii gene containing an open reading frame of 333 nucleotides encoding a 12.7 kDa protein. The novel protein, DIP13, exhibits 60% identity with two mammalian proteins, human NA14 and an unnamed mouse protein. Homologous sequences are also present in several protozoan, trematode and fish genomes, but no homologs have been found in the completed genomes of yeast, Drosophila, C. elegans and A. thaliana. By using a specific antibody we have localized DIP13 to microtubule structures, namely basal bodies, flagellar axonemes and cytoplasmic microtubules. Anti-DIP13 antibody also specifically recognized human NA14 by immunofluorescence and stained basal bodies and flagella of human sperm cells as well as the centrosome of HeLa cells. Expression of the DIP13 open reading frame in antisense orientation in Chlamydomonas resulted in multinucleate, multiflagellate cells, which suggests a role for this protein in ensuring proper cell division. Thus, DIP13/NA14 could represent the founding members of a new class of highly conserved proteins that are associated with microtubule structures. PMID- 12640031 TI - The scaffold protein IB1/JIP-1 is a critical mediator of cytokine-induced apoptosis in pancreatic beta cells. AB - In insulin-secreting cells, cytokines activate the c-Jun N-terminal kinase (JNK), which contributes to a cell signaling towards apoptosis. The JNK activation requires the presence of the murine scaffold protein JNK-interacting protein 1 (JIP-1) or human Islet-brain 1(IB1), which organizes MLK3, MKK7 and JNK for proper signaling specificity. Here, we used adenovirus-mediated gene transfer to modulate IB1/JIP-1 cellular content in order to investigate the contribution of IB1/JIP-1 to beta-cell survival. Exposure of the insulin-producing cell line INS 1 or isolated rat pancreatic islets to cytokines (interferon-gamma, tumor necrosis factor-alpha and interleukin-1beta) induced a marked reduction of IB1/JIP-1 content and a concomitant increase in JNK activity and apoptosis rate. This JNK-induced pro-apoptotic program was prevented in INS-1 cells by overproducing IB1/JIP-1 and this effect was associated with inhibition of caspase 3 cleavage. Conversely, reducing IB1/JIP-1 content in INS-1 cells and isolated pancreatic islets induced a robust increase in basal and cytokine-stimulated apoptosis. In heterozygous mice carrying a selective disruption of the IB1/JIP-1 gene, the reduction in IB1/JIP-1 content in happloinsufficient isolated pancreatic islets was associated with an increased JNK activity and basal apoptosis. These data demonstrate that modulation of the IB1-JIP-1 content in beta cells is a crucial regulator of JNK signaling pathway and of cytokine induced apoptosis. PMID- 12640032 TI - Cadherin-mediated adhesion is essential for myofibril continuity across the plasma membrane but not for assembly of the contractile apparatus. AB - The strong coordinated contraction of heart muscle is dependent on the correct alignment and connection of the myofibrils across the plasma membrane. Previous studies indicate that N-cadherin is involved in cardiac myocyte adhesion and myofibrillogenesis. To investigate whether N-cadherin is specifically required for normal myocyte structure and function, we cultured myocytes from wild-type, N cadherin-null and mutant embryos expressing the epithelial cadherin E-cadherin. In contrast to previous studies in chicken using N-cadherin-perturbing antibodies, our in vitro studies with mouse cells demonstrate that N-cadherin is not required for myofibrillogenesis, but is critical for myofibril organization. That is, N-cadherin-deficient myocytes beat and myofibrils were well formed; however, alignment of the myofibrils through regions of cell-cell contact was lost, resulting in their random orientation. Gap junctions were perturbed in the N-cadherin-null myocytes. By contrast, focal contacts appeared normal in the mutant cells. Furthermore, E-cadherin restored normal cell morphology and behavior to the N-cadherin-deficient myocytes, including proper alignment of the myofibrils. We conclude that a different adhesive system, most probably integrin, is responsible for myofibrillogenesis in the N-cadherin-null myocytes. PMID- 12640033 TI - Direct correlation of collagen matrix deformation with focal adhesion dynamics in living corneal fibroblasts. AB - The purpose of this study was to develop and apply a new model for investigating how the organization and movement of cell-matrix adhesion sites correlate with force generation by corneal fibroblasts on a fibrillar collagen extracellular matrix. Primary cultures of rabbit corneal fibroblasts were transfected using a vector encoding GFP-zyxin to allow visualization of adhesion sites. Cells were plated at low density on top of 100 micro m thick fibrillar collagen lattices embedded with 2 micro m diameter red fluorescent beads. Time-lapse imaging was performed at one minute intervals for up to 3 hours. At each time interval, GFP zyxin, bead and DIC images were acquired in rapid succession using filter wheels. Cells were treated with cytochalasin D and/or Triton X-100 at the end of each experiment. The movements of adhesions and nearby matrix landmarks were measured and correlated from the time-lapse digital images, and the size, intensity and orientation of the adhesions were quantified. GFP-zyxin was detected in adhesions of transfected corneal fibroblasts as confirmed using vinculin counterstaining. Time-lapse imaging revealed extensions and retractions of cell processes and displacements of the fiduciary beads that were similar to control cells. Extending processes exhibited the most complex behavior, with new adhesions continuously forming at the leading edge while existing adhesions moved backward in a retrograde fashion. This process generated tractional forces as indicated by pulling in of the extracellular matrix in front of the cell. Interestingly, during extension, adhesions along the ventral surface of the cell body generally moved toward those at the tip, resulting in contractile-like shortening and matrix compression at the base of lamellipodia. Overall, a high correlation was found between both the magnitude (R=0.87, P<0.001) and direction (R=0.98, P<0.001) of the adhesions and nearby matrix displacements. Cytochalasin D induced rapid and reversible disassembly of adhesions, cell elongation and matrix relaxation, including decompression at the base of the lamellipodia. This new experimental model allows direct, dynamic assessment of cell-matrix interactions on a fibrillar collagen matrix. Our results are consistent with the previously described 'frontal towing' model of cell motility and demonstrate for the first time that this mechanism is employed by cells interacting with a fibrillar extracellular matrix. PMID- 12640034 TI - A p50-like Y-box protein with a putative translational role becomes associated with pre-mRNA concomitant with transcription. AB - In vertebrates free messenger ribonucleoprotein (RNP) particles and polysomes contain an abundant Y-box protein called p50 (YB-1), which regulates translation, presumably by affecting the packaging of the RNA. Here, we have identified a p50 like protein in the dipteran Chironomus tentans and studied its relation with the biogenesis of mRNA in larval salivary glands. The salivary gland cells contain polytene chromosomes with the transcriptionally active regions blown up as puffs. A few giant puffs, called Balbiani rings (BRs), generate a transcription product, a large RNP particle, which can be visualised (with the electron microscope) during its assembly on the gene and during its transport to and through the nuclear pores. The p50-like protein studied, designated Ct-p40/50 (or p40/50 for short), was shown to contain a central cold-shock domain, an alanine- and proline rich N-terminal domain, and a C-terminal domain with alternating acidic and basic regions, an organisation that is characteristic of p50 (YB-1). The p40/50 protein appears in two isoforms, p40 and p50, which contain 264 and 317 amino acids, respectively. The two isoforms share the first 258 amino acids and thus differ in amino-acid sequence only in the region close to the C-terminus. When a polyclonal antibody was raised against p40/50, western blot analysis and immunocytology showed that p40/50 is not only abundant in the cytoplasm but is also present in the nucleus. Immunolabelling of isolated polytene chromosomes showed that p40/50 appears in transcriptionally active regions, including the BRs. Using immunoelectron microscopy we revealed that p40/50 is added along the nascent transcripts and is also present in the released BR RNP particles in the nucleoplasm. Finally, by UV crosslinking in vivo we showed that p40/50 is bound to both nuclear and cytoplasmic poly(A) RNA. We conclude that p40/50 is being added cotranscriptionally along the growing BR pre-mRNA, is released with the processed mRNA into the nucleoplasm and probably remains associated with the mRNA both during nucleocytoplasmic transport and protein synthesis. Given that the p40/p50 protein, presumably with a role in translation, is loaded onto the primary transcript concomitant with transcription, an early programming of the cytoplasmic fate of mRNA is indicated. PMID- 12640035 TI - Essential role of the C. elegans Arp2/3 complex in cell migration during ventral enclosure. AB - Migration of cells through the reorganization of the actin cytoskeleton is essential for morphogenesis of multicellular animals. In a cell culture system, the actin-related protein (Arp) 2/3 complex functions as a nucleation core for actin polymerization when activated by the members of the WASP (Wiskott-Aldrich syndrome protein) family. However, the regulation of cell motility in vivo remains poorly understood. Here we report that homologues of the mammalian Arp2/3 complex and N-WASP in Caenorhabditis elegans play an important role in hypodermal cell migration during morphogenesis, a process known as ventral enclosure. In the absence of one of any of the C. elegans Arp2/3 complex subunits (ARX-1, ARX-2, ARX-4, ARX-5, ARX-6 or ARX-7) or of N-WASP (WSP-1), hypodermal cell migration led by actin-rich filopodia formation is inhibited during ventral enclosure owing to the reduction of filamentous actin formation. However, there is no effect on differentiation of hypodermal cells and dorsal intercalation. Disruption of the function of ARX-1 and WSP-1 in hypodermal cells also resulted in hypodermal cell arrest during ventral enclosure, suggesting that their function is cell autonomous. WSP-1 protein activated Arp2/3-mediated actin polymerization in vitro. Consistent with these results, the Arp2/3 complex and WSP-1 colocalized at the leading edge of migrating hypodermal cells. The stable localization of WSP-1 was dependent on the presence of Arp2/3 complex, suggesting an interaction between the Arp2/3 complex and WSP-1 in vivo. PMID- 12640036 TI - cAMP-induced AQP2 translocation is associated with RhoA inhibition through RhoA phosphorylation and interaction with RhoGDI. AB - We have recently demonstrated that inhibition of Rho GTPase with Clostridium difficile toxin B, or with Clostridium botulinum C3 toxin, causes actin depolymerization and translocation of aquaporin 2 (AQP2) in renal CD8 cells in the absence of hormonal stimulation. Here we demonstrate that Rho inhibition is part of the signal transduction cascade activated by vasopressin leading to AQP2 insertion into the apical membrane. Quantitation of active RhoA (GTP-bound) by selective pull down experiments demonstrated that the amount of active RhoA decreased upon stimulation of CD8 cells with the cAMP-elevating agent forskolin. Consistent with this observation, forskolin treatment resulted in a decreased expression of membrane-associated (active) Rho, as assessed by cell fractionation followed by western blotting analysis. In addition, the abundance of the endogenous Rho GDP dissociation inhibitor (Rho-GDI) was found to have decreased in the membrane fraction after forskolin stimulation. Co-immunoprecipitation experiments revealed that, after forskolin stimulation, the amount of Rho-GDI complexed with RhoA increased, suggesting that Rho GTPase inhibition occurs through association of RhoA with Rho-GDI. Finally, forskolin stimulation was associated with an increase in Rho phosphorylation on a serine residue, a protein modification known to stabilize the inactive form of RhoA and to increase its interaction with Rho-GDI. Taken together, these data demonstrate that RhoA inhibition through Rho phosphorylation and interaction with Rho-GDI is a key event for cytoskeletal dynamics controlling cAMP-induced AQP2 translocation. PMID- 12640037 TI - Evidence that Ca(2+) cycling by the plasma membrane Ca(2+)-ATPase increases the 'excitability' of the extracellular Ca(2+)-sensing receptor. AB - The extracellular Ca(2+)-sensing receptor (CaR) is a widely expressed G-protein coupled receptor that translates information about [Ca(2+)] in the extracellular milieu to the interior of the cell, usually via intracellular Ca(2+) signaling pathways. Using fura-2 imaging of cytoplasmic [Ca(2+)], we observed that HEK293 cells expressing CaR oscillated readily under conditions permissive for CaR activation. Spiking was also triggered in the absence of external Ca(2+) by the CaR agonist spermine (1 mM). Oscillating cells were typically located in clusters of closely apposed cells, but Ca(2+) spiking was insensitive to the gap junction inhibitor 18alpha-glycyrrhetinic acid. We hypothesized that Ca(2+) signals might be amplified, in part, through a positive feedback loop in which Ca(2+) extrusion via the plasma membrane Ca(2+)-ATPase (PMCA) activates CaRs on the same cell or adjacent cells through local increases in [Ca(2+)](out). In support of this idea, addition of exogenous Ca(2+) buffers (keeping free [Ca(2+)](out) constant) attenuated or eliminated Ca(2+) signals (manifested as oscillations), as did PMCA inhibitors (HgCl(2), orthovanadate and Caloxin 2A1). Measurement of extracellular [Ca(2+)] using the near membrane probe fura-C(18) revealed that external [Ca(2+)] rose following receptor activation, sometimes displaying an oscillatory pattern. Our data suggest that PMCA-mediated cycling of Ca(2+) across the plasma membrane leads to localized increases in [Ca(2+)](out) that increase the excitability of CaR. PMID- 12640038 TI - Expression in Xenopus oocytes shows that WT1 binds transcripts in vivo, with a central role for zinc finger one. AB - The Wilms' tumour suppressor gene WT1 encodes a protein involved in urogenital development and disease. The salient feature of WT1 is the presence of four 'Kruppel'-type C(2)-H(2) zinc fingers in the C-terminus. Uniquely to WT1, an evolutionarily conserved alternative splicing event inserts three amino acids (KTS) between the third and fourth zinc fingers, which disrupts DNA binding. The ratio of +KTS:-KTS isoforms is crucial for normal development. Previous work has shown that WT1 (+KTS) interacts with splice factors and that WT1 zinc fingers, particularly zinc finger one, bind to RNA in vitro. In this study we investigate the role of zinc finger one and the +KTS splice in vivo by expressing tagged proteins in mammalian cells and Xenopus oocytes. We find that both full-length +/ KTS isoforms and deletion constructs that include zinc finger one co-sediment with ribonucleoprotein particles (RNP) on density gradients. In Xenopus oocytes both isoforms located to the lateral loops of lampbrush chromosomes. Strikingly, only the +KTS isoform was detected in B-snurposomes, but not when co-expressed with -KTS. However, co-expression of the C-terminus (amino acids 233-449, +KTS) resulted in snurposome staining, which is consistent with an in vivo interaction between isoforms via the N-terminus. Expressed WT1 was also detected in the RNA rich granular component of nucleoli and co-immunoprecipitated with oocyte transcripts. Full-length WT1 was most stably bound to transcripts, followed by the C-terminus; the least stably bound was CTDeltaF1 (C-terminus minus zinc finger one). Expression of the transcription factor early growth response 1 (EGR1), whose three zinc fingers correspond to WT1 zinc fingers 2-4, caused general chromosomal loop retraction and transcriptional shut-down. However, a construct in which WT1 zinc finger one was added to EGR1 mimicked the properties of WT1 (-KTS). We suggest that in evolution, WT1 has acquired the ability to interact with transcripts and splice factors because of the modification of zinc finger one and the +KTS alternative splice. PMID- 12640039 TI - PARP-3 localizes preferentially to the daughter centriole and interferes with the G1/S cell cycle progression. AB - A novel member of the poly(ADP-ribose) polymerase (PARP) family, hPARP-3, is identified here as a core component of the centrosome. hPARP-3 is preferentially localized to the daughter centriole throughout the cell cycle. The N-terminal domain (54 amino acids) of hPARP-3 is responsible for its centrosomal localization. Full-length hPAPR-3 (540 amino acids, with an apparent mass of 67 kDa) synthesizes ADP-ribose polymers during its automodification. Overexpression of hPARP-3 or its N-terminal domain does not influence centrosomal duplication or amplification but interferes with the G1/S cell cycle progression. PARP-1 also resides for part of the cell cycle in the centrosome and interacts with hPARP-3. The presence of both PARP-1 and PARP-3 at the centrosome may link the DNA damage surveillance network to the mitotic fidelity checkpoint. PMID- 12640040 TI - Genome-wide expression screens indicate a global role for protein kinase CK2 in chromatin remodeling. AB - Protein kinase CK2, a vital, pleiotropic and highly conserved serine/threonine phosphotransferase is involved in transcription-directed signaling, gene control and cell cycle regulation and is suspected to play a role in global processes. Searching for these global roles, we analyzed the involvement of CK2 in gene expression at cell cycle entry by using genome-wide screens. Comparing expression profiles of Saccharomyces cerevisiae wild-type strains with strains with regulatory or catalytic subunits of CK2 deleted, we found significant alterations in the expression of genes at all cell cycle phases and often in a subunit- and isoform-specific manner. Roughly a quarter of the genes known to be regulated by the cell cycle are affected. Functionally, the genes are involved with cell cycle entry, progression and exit, including spindle pole body formation and dynamics. Strikingly, most CK2-affected genes exhibit no common transcriptional control features, and a considerable proportion of temporarily altered genes encodes proteins involved in chromatin remodeling and modification, including chromatin assembly, (anti-)silencing and histone (de-)acetylation. In addition, various metabolic pathway and nutritional supply genes are affected. Our data are compatible with the idea that CK2 acts at different levels of cellular organization and that CK2 has a global role in transcription-related chromatin remodeling. PMID- 12640041 TI - Aspergillus fumigatus conidia survive and germinate in acidic organelles of A549 epithelial cells. AB - Aspergillus fumigatus is an environmental mould that can cause invasive disease in the immunocompromised host. Previous work has shown that conidia can be internalized by lung epithelial cells (A549) and murine macrophages (J774) in vitro. Therefore, the purpose of this study was to determine the fate of A. fumigatus conidia within the endosomal network of these cells. Co-localization of conidia expressing green fluorescent protein with proteins present in the early endosomal (CD71) and lysosomal (CD63, LAMP-1) membrane was assessed using confocal microscopy. In J774 cells, 75% of internalized conidia were found in phagosomes containing LAMP-1 120 minutes post-infection. In A549 cells, 55% and 58% of internalized conidia were found to co-localize with LAMP-1 and CD63 by 24 hours. Cathepsin D also co-localized with internalized conidia in A549 cells. Phagosomes containing conidia were shown to be acidified in both cell types. Less than 1% of the initial inoculum survived in J774 cells by 12 hours post infection. After 24 hours, 3% of internalized conidia survived in A549 cells and 34% of these had germinated. By 36 hours, the germlings were able to escape the phagosome and form extracellular hyphae without lysis of the host cell. PMID- 12640042 TI - The block of ryanodine receptors selectively inhibits fetal myoblast differentiation. AB - Differentiation and morphogenesis of skeletal muscle are complex and asynchronous events that involve various myogenic cell populations and extracellular signals. Embryonic and fetal skeletal myoblasts are responsible for the formation of primary and secondary fibers, respectively, although the mechanism that diversifies their fate is not fully understood. Calcium transients appear to be a signaling mechanism that is widely utilized in differentiation and embryogenesis. In mature skeletal muscle, calcium transients are generated mainly by ryanodine receptors (type 1 and type 3), which are involved in excitation-contraction coupling. However, it is not clear whether the activity of these receptors is important for contractile activity alone or whether it may also play a role in regulating the differentiation/developmental processes. To clarify this point, we first examined the expression of the receptors during development. The results show that the expression of both receptors appears as early as E13 during limb muscle development and parallels the expression of skeletal myosin. The expression and the activity of both receptors is maintained in vitro by all myogenic cell populations isolated from different stages of development, including somitic, embryonic and fetal myoblasts and satellite cells. Blocking ryanodine receptor activity by using ryanodine inhibits in vitro differentiation of fetal myoblasts (judged by the expression of sarcomeric myosin and formation of multinucleated myotubes) but not of somitic or embryonic and satellite muscle cells. This block is caused by the transcriptional inhibition of markers characteristic of terminal differentiation, rather than commitment, as the expression of muscle regulatory factors is not impaired by ryanodine treatment. Taken together, the data reported in this paper demonstrate that, although calcium transients represent a general mechanism for the control of differentiation and development, multiple calcium-dependent pathways may be relevant in different myogenic populations during development. Moreover, since fetal myoblasts are responsible for the formation of secondary fibers during development, and therefore for the building of the bulk of muscular mass, these results suggest that calcium release from ryanodine receptors plays a role in the histogenesis of mammalian skeletal muscle. PMID- 12640043 TI - A novel endocytic pathway induced by clustering endothelial ICAM-1 or PECAM-1. AB - Antibody conjugates directed against intercellular adhesion molecule (ICAM-1) or platelet-endothelial cell adhesion molecule (PECAM-1) have formed the basis for drug delivery vehicles that are specifically recognized and internalized by endothelial cells. There is increasing evidence that ICAM-1 and PECAM-1 may also play a role in cell scavenger functions and pathogen entry. To define the mechanisms that regulate ICAM-1 and PECAM-1 internalization, we examined the uptake of anti-PECAM-1 and anti-ICAM-1 conjugates by endothelial cells. We found that the conjugates must be multimeric, because monomeric anti-ICAM-1 and anti PECAM-1 are not internalized. Newly internalized anti-ICAM-1 and anti-PECAM-1 conjugates did not colocalize with either clathrin or caveolin, and immunoconjugate internalization was not reduced by inhibitors of clathrin mediated or caveolar endocytosis, suggesting that this is a novel endocytic pathway. Amiloride and protein kinase C (PKC) inhibitors, agents known to inhibit macropinocytosis, reduced the internalization of clustered ICAM-1 and PECAM-1. However, expression of dominant-negative dynamin-2 constructs inhibited uptake of clustered ICAM-1. Binding of anti-ICAM-1 conjugates stimulated the formation of actin stress fibers by human umbilical vein endothelial cells (HUVEC). Latrunculin, radicicol and Y27632 also inhibited internalization of clustered ICAM-1, suggesting that actin rearrangements requiring Src kinase and Rho kinase (ROCK) were required for internalization. Interestingly, these kinases are part of the signal transduction pathways that are activated when circulating leukocytes engage endothelial cell adhesion molecules, suggesting the possibility that CAM-mediated endocytosis is regulated using comparable signaling pathways. PMID- 12640044 TI - Migration of adult myogenic precursor cells as revealed by GFP/nLacZ labelling of mouse transplantation chimeras. AB - We studied the migratory behaviour of adult muscle precursor cells in the mouse into and from skeletal muscle grafts using green fluorescent protein (GFP) and nuclear LacZ transgenes as complementary and double markers of the cell's origin. Owing to the small molecular mass and extreme solubility of GFP, this label provided a drastically increased sensitivity for detection compared with the markers that had been used previously. During the first six weeks after the operation, the graft/host border was well defined, with only occasional local intermingling and co-fusion of host and donor myogenic cells. Seven to eleven weeks after the operation we found that the host myogenic cells had migrated into the graft, and graft myogenic cells had migrated into the adjacent host muscle, with integration of donor nuclei into pre-existing myotubes or muscle fibres. There was no indication of an origin of, or target for, these myogenic cells besides neighbouring muscles. Our observations indicate migration of these cells through solid muscle tissue, over a distance of several millimetres. The migratory activity of adult myogenic precursor cells can be stimulated by traumatic events in either the target muscle or the muscle of origin. PMID- 12640045 TI - Mechanics of cell spreading: role of myosin II. AB - As it migrates over a substratum, a cell must exert different kinds of forces that act at various cellular locations and at specific times. These forces must therefore be coordinately regulated. The Rho-family GTPases Rac1 and Cdc42 promote actin polymerization that drives extension of the leading cell edge. Subsequently, RhoA regulates myosin-dependent contractile force, which is required for formation of adhesive contacts and stress fibers. During cell spreading, however, the activity of RhoA is reduced by a mechanism involving the tyrosine kinases c-Src and focal adhesion kinase (FAK), and the p190RhoGAP. It has been proposed that this reduction of RhoA activity facilitates edge extension by reducing myosin-dependent contractile forces that could resist this process. We have directly tested this hypothesis by correlating myosin activity with the rate of cell spreading on a substratum. The rate of spreading is inversely related to the myosin activity. Furthermore, spreading is inhibited by low concentrations of cytochalasin D, as expected for a process that depends on the growth of uncapped actin filaments. Cell indentation measurements show that a myosin-dependent viscoelastic force resists cell deformation. PMID- 12640046 TI - Central-pair-linked regulation of microtubule sliding by calcium in flagellar axonemes. AB - The movement of eukaryotic flagella and cilia is regulated by intracellular calcium. We have tested a model in which the central pair of microtubules mediate the effect of Ca(2+) to modify the dynein activity. We used a novel microtubule sliding assay that allowed us to test the effect of Ca(2+) in the presence or absence of the central-pair microtubules. When flagellar axonemes of sea-urchin sperm were exposed to ATP in the presence of elastase, they showed different types of sliding disintegration depending on the ATP concentration: at low concentrations of ATP (/=100 micro M), a large proportion of the axonemes showed limited sliding and split lengthwise into a pair of two microtubule bundles, one of which was thicker than the other. The sliding behaviour of the axonemes was also influenced by Ca(2+). Thus, at 1 mM ATP, the proportion of axonemes that split into two bundles increased from 25% at <10(-9) M Ca(2+) to 60% at 10(-4) M Ca(2+), whereas the sliding velocity of doublets during the splitting did not change. Electron microscopy of split bundles showed that the thicker bundles contained five or six doublets and the central pair, whereas the thinner bundles contained three or four doublets but not the central pair. Closer examinations revealed that the thicker bundles were dominated by four patterns of doublet combinations: doublets 8-9-1-2-3-4, 8-9-1-2-3, 4-5-6-7-8 and 3-4-5-6-7-8. This indicates that the sliding occurred preferentially at one or two fixed interdoublet sites on either side of the central-pair microtubules, whereas the sliding at the remaining interdoublet sites was inhibited under these conditions. Ca(2+) reduced the appearance of the 4-5-6-7-8 and 3-4-5-6-7-8 patterns and increased the 8-9-1-2-3-4 and 8-9-1-2-3 patterns. The splitting patterns are possibly related to the switching mechanism of the dynein activity underlying the cyclical flagellar bending. To investigate the role of the central pair in the regulation of the dynein activity by Ca(2+), we studied the behaviour of singlet microtubules applied to the dynein arms exposed on the doublets of the split bundles that were either associated with the central pair or not. Microtubules moved along both the thicker and the thinner bundles but the frequency of microtubule sliding on the thinner (i.e. the central-pair-less) bundles was three to four times (at 62 or < 62 years) and disease severity (H-Y > III or H-Y < or = II), the median H/M ratio of the older and more severe group was significantly lower than that of the younger and less severe group (p = 0.005). CONCLUSION: This study suggests that late onset, high severity PD is associated with myocardial sympathetic dysfunction. PMID- 12640056 TI - A note on Hoover's sign. PMID- 12640055 TI - Limb length and dementia in an older Korean population. AB - OBJECTIVES: There has been little research into risk factors for dementia outside Western settings, in particular the importance of early life nutrition as estimated by adult body size. This study investigated the associations of arm and leg length with cognitive impairment and dementia in a community sample of older Korean people. METHODS: 746 community residents aged 65 or over were clinically assessed for dementia and cognitive impairment. The following were also measured: arm length (demispan), leg length (iliac crest height), and sitting height (standing height minus iliac crest height). Reproductive history was also ascertained in women. RESULTS: Shorter demispan and leg length were associated with increased age and lower education. They were also associated with dementia and Alzheimer's disease after adjustment for these factors. These associations were only significant in women but were not explained substantially by timing of the menarche or menopause. The association between lower education and dementia was also stronger in women, but was not explained substantially by limb length. CONCLUSIONS: Shorter limb length was associated with lower childhood socioeconomic status, as estimated by the presence/duration of formal education. It was also independently associated with dementia and Alzheimer's disease. Sex differences in this association might be explained by gender disadvantage in early life for this cohort or by different associations with health states (for example, cardiovascular disease) later in life. PMID- 12640058 TI - One year changes in disability in multiple sclerosis: neurological examination compared with patient self report. AB - OBJECTIVE: To characterise prospectively the relation between one year changes in neurologist rating of neurological exam abnormalities as measured by the Expanded Disability Status Scale (EDSS) and changes in patient perceived disability as measured by the Guy's Neurological Disability Scale (GNDS) in patients with multiple sclerosis. METHODS: Two hundred and fifty patients with MS were recruited at an outpatient clinic. Disability at baseline and one year follow-up was assessed using the EDSS and GNDS. Correlations between change in EDSS, GNDS sum score, Functional Systems and GNDS subcategories were studied as well as the significance of changes in EDSS associated with changes in perceived disability. RESULTS: The correlation between one year changes in EDSS versus GNDS was substantially lower (0.19) than cross-sectional correlations between EDSS and GNDS either at baseline (0.62) or at follow-up (0.77). Notably, changes in functional system scores that are based on neurological examination are poorly or not at all correlated with changes in disability as perceived by the patient. Analysing the impact of a significant worsening in EDSS-score, a commonly applied outcome criterion in clinical trials, we found that this was associated with significant worsening, insignificant change, and significant improvement in the patients' perceived disability in 45%, 39% and 15% of patients, respectively. CONCLUSION: Patients' perception of change in disability differs not only quantitatively but also qualitatively from that of an examining physician. This seems to be due both to the fact that there are true differences in change as perceived by the patient and that measured by the physician and to the fact that changes in many dimensions of disability that are relevant to the patient have no measurable impact on the EDSS. PMID- 12640057 TI - Neuropsychological prediction of conversion to dementia from questionable dementia: statistically significant but not yet clinically useful. AB - BACKGROUND: Verbal memory impairment, one of the earliest signs of Alzheimer's disease (AD), may help identify people with cognitive impairment, insufficient for a diagnosis of dementia (questionable dementia: QD), at risk of developing AD. Other cognitive parameters have been found that may indicate which people with QD will go on to develop dementia. Nevertheless, some researchers have reported only partial success in differentiating between mild AD and age related cognitive impairment. OBJECTIVES: To discover if there are early, pre-clinical cognitive markers that could help identify patients attending our memory clinic who were at risk of developing dementia. METHODS: Multidisciplinary assessment of a consecutive sample of 195 patients with QD seen in a National Health Service hospital outpatient clinic; 135 seen for a mean follow up of 24.5 months. RESULTS: Conversion rate to dementia was 27.4% (37 of 135). A diagnosis of probable or possible AD was made in 15.6% (21 of 135) of cases. Despite statistically significant differences in some cognitive tasks between those who did and those who did not go on to dement, Cox regression analyses failed to improve prediction rates markedly above base rates and were unstable. CONCLUSION: A large number of studies claim good prediction of conversion to dementia using cognitive test scores. Although this study produced similarly good sensitivity and specificity values, proper consideration of the statistical analyses and their clinical significance suggested that these prediction methods are currently too imprecise for clinical use. Use of cognitive indicators combined with neuroradiological, neuropathological, and genetic factors for predicting conversion to dementia might prove more reliable but may be beyond the scope of many geriatric services. PMID- 12640060 TI - Cognitive impairment in probable multiple sclerosis. AB - OBJECTIVES: To evaluate and characterise cognitive impairment in the very early stage of multiple sclerosis (MS), in which patients are still diagnosed as suffering from probable MS. METHODS: The Brief Repeatable Battery Neuropsychological (BRB-N) that has been validated for MS patients was used. Abnormal performance was defined as one standard deviation below the mean reported for healthy age matched subjects. Neurological disability and brain magnetic resonance imaging (MRI) were performed for all patients. Correlation coefficients were calculated between disease burden variables and performance on the BRB-N. RESULTS: Sixty seven patients with probable MS were evaluated within a mean of one month of the onset of new neurological symptoms. Evidence for the presence of cognitive impairment was shown in 53.7% of patients. Verbal abilities and attention span were most frequently affected. Impairment was not correlated with neurological disability or MRI disease burden. CONCLUSION: Prevalent cognitive impairment already exists at onset of MS. PMID- 12640061 TI - Reliability of the variables in a new set of models that predict outcome after stroke. AB - OBJECTIVES: To provide valid predictions of outcome, the variables included in a prognostic model must be capable of reliable collection. The authors have recently reported a set of simple but rigorously developed models that predict outcome after stroke. The aim of this study was to establish the inter-rater reliability of the variables included in the models. METHODS: Inter-rater agreement was measured prospectively (between two clinicians; 92 patients) and retrospectively (between two auditors; 200 patients) and the validity of the data collected retrospectively was estimated by comparing them with data collected prospectively (195 patients). In the prospective study inter-rater agreement for urinary incontinence and for the variables of three other previously published models was also measured. The median difference (md) between ages and kappa statistics for other variables was calculated. RESULTS: For the model variables, prospective agreement ranged from good to excellent (age: md 0 years; living alone before the stroke kappa 0.84; pre-stroke functional independence kappa 0.67; normal verbal Glasgow Coma Scale score kappa 0.79; ability to lift both arms against gravity kappa 0.97; ability to walk unaided kappa 0.91) while retrospective agreement (age: md 0 years; kappa 0.55-0.92) and agreement between prospective and retrospective observers (age: md 0 years; kappa 0.49-0.78) was acceptable but less good. Prospective agreement was excellent for urinary incontinence (kappa 0.87) and variable for the other models (kappa 0.23-0.81) CONCLUSION: The variables included in these new simple models of outcome after stroke are capable of reliable collection, comparable to or better than that of the other predictive variables considered. When collected retrospectively, the model variables are likely to remain reliable and reasonably valid. PMID- 12640062 TI - Visible infarction on computed tomography is an independent predictor of poor functional outcome after stroke, and not of haemorrhagic transformation. AB - OBJECTIVES: To examine a very large dataset to provide a robust answer to the question of whether visible infarction on computed tomography was (a) an independent predictor of functional outcome at all times up to 48 hours after stroke, and (b) independently associated with haemorrhagic transformation, with or without antithrombotic treatment. METHODS: The study assessed associations between visible infarction, time to randomisation, baseline neurological deficit, stroke syndrome, allocated aspirin or heparin treatment, recurrent haemorrhagic stroke, early death and six month functional outcome in the International Stroke Trial. RESULTS: Of 12 550 patients, 6267 (50%) had visible infarction up to 48 hours after stroke. The prevalence of visible infarction increased with increasing time from onset and extent of the stroke syndrome. Visible infarction was independently associated with increased death within 14 days (odds ratio (OR) 1.17, 95% CI 1.02 to 1.35), and of death or dependency at six months (OR 1.42, 95% CI 1.31 to 1.55), an absolute increase of 13%, or 130 per 1000 more dead or dependent patients with visible infarction than without it. There was no significant independent relation between visible infarction and fatal or non fatal haemorrhagic transformation, or interaction between visible infarction and aspirin or heparin treatment allocation with six month functional outcome. CONCLUSIONS: Visible infarction on computed tomography up to 48 hours after stroke is an independent adverse prognostic sign. PMID- 12640063 TI - Recanalisation of cerebral venous thrombosis. AB - OBJECTIVE: To investigate recanalisation in the first 12 months after cerebral venous thrombosis. METHODS: 33 consecutive patients presenting with cerebral venous thrombosis were enrolled in the study. Diagnosis was made by magnetic resonance imaging (MRI) and magnetic resonance venography (MRV) or catheter angiography. Patients were initially treated with intravenous heparin. Warfarin was given for at least four months. Cerebral MRI and MRV were done at four months and repeated after 12 months if venous thrombosis persisted. Outcome was evaluated by the Rankin scale at 12 months. RESULTS: Outcome at 12 months was good, with a median modified Rankin scale score of 0 (range 0 to 2); 27 patients (82%) had no residual deficits. No patient suffered recurrent cerebral venous thrombosis, deep vein thrombosis, or pulmonary embolism during follow up. After four months, all deep cerebral veins and cavernous sinuses, 94% of superior sagittal sinuses, 80% of straight sinuses, 73% of jugular veins, 58% of transverse sinuses, and 41% of sigmoid sinuses had recanalised. No further recanalisation was observed thereafter. CONCLUSIONS: The results suggest that recanalisation only occurs within the first four months following cerebral venous thrombosis and not thereafter, irrespective of oral anticoagulation. PMID- 12640064 TI - Specific phobia is a frequent non-motor feature in stiff man syndrome. AB - OBJECTIVE: To investigate systematically the rate and type of phobia in stiff man syndrome and its variants, and to compare patients with stiff man syndrome with and without phobia for sociodemographic and neurological characteristics. METHODS: 43 consecutive patients with stiff man syndrome referred to a university department of neurology were assessed using the anxiety disorders interview schedule, revised (ADIS-R), a structured diagnostic interview for anxiety disorders, in addition to a full clinical neurological and psychiatric assessment. RESULTS: 19 patients (44.2%) developed task specific phobia--that is, fear and avoidance of situations difficult to master owing to the motor symptoms of stiff man syndrome (such as crossing streets). Three further patients (7%) had subthreshold phobia--that is, phobic anxiety without avoidance. There were no significant differences between patients with and without phobia in terms of age, illness duration, type of stiff man syndrome, antibody status, or frequency of falls. Patients with phobia were more likely to present with exaggerated startle responses and to have an initial misdiagnosis of psychogenic movement disorder. CONCLUSIONS: Specific phobia is a frequent non-motor symptom of stiff man syndrome. Early recognition is an important aid to correct diagnosis. The aetiology of phobia in stiff man syndrome is unknown. There is no evidence of a direct pathogenic role of autoantibodies directed against glutamic acid decarboxylase in the development of phobia. PMID- 12640065 TI - Clinical application of neuroimaging in epilepsy. AB - OBJECTIVE: To evaluate the use of neuroimaging in clinical practice and to assess the prevalence of detected structural abnormalities in epilepsy patients in a clinical set up. METHODS: 919 outpatients were identified and the scan results reviewed. A total of 677 patients had chronic active epilepsy (88 had idiopathic generalised epilepsy (IGE), 588 had localisation related epilepsy, one had symptomatic generalised epilepsy), 57 had a single epileptic seizure, 46 were in remission, and 139 had non-epileptic attacks. RESULTS: 391 patients had no scan (53 patients in this group had IGE, 182 had localisation related epilepsy, one had generalised symptomatic epilepsy, 18 had single epileptic attacks, 21 were in remission, 116 had non-epileptic attacks). Altogether 528 patients had a scan, the results were not available in 33, 163 had x ray computed tomography (CT) only, 178 had standard magnetic resonance imaging (MRI) (slice thickness 5 mm), and 154 had high resolution MRI (including a T1 weighted sequence with 1.5 mm thick slices). Some 252 of 495 scans (51%) were abnormal. Abnormalities were hippocampal sclerosis (n=128), atrophy or non-specific white matter lesions (n=35), vascular abnormalities (n=27), tumours (n=25), brain damage (n=24), malformations of cortical development (n=13). Excluding atrophy and non-specific white matter lesions the prevalence of detected abnormalities was 54% in localisation related epilepsy, 18% in single seizure patients, 16% in epilepsy in remission, and 0% in IGE and non-epileptic attacks. CONCLUSIONS: Abnormalities were detected in more than half of all patients with localisation related epilepsy, and in about one in five patients with single seizures or epilepsy in remission. Many patients had no scan or only CT or standard MRI. The true prevalence of structural abnormalities may be have been higher. Scanning did not add any information in patients with IGE or non-epileptic attacks. PMID- 12640066 TI - Preoperative motor system brain mapping using positron emission tomography and statistical parametric mapping: hints on cortical reorganisation. AB - OBJECTIVES: This study investigated the applicability of statistical parametric mapping (SPM) for analysing individual preoperative brain mapping studies in patients with cerebral mass lesions for neurosurgical planning. The study further investigated if hints on functional reorganisation processes can be found. METHODS: Nine adult patients with cerebral mass lesions underwent activation [(15)O]water-PET under stimulation by finger (n=9) and foot (n=4) movement. Individual SPM-t-maps were computed without anatomical normalisation and coregistered to the individual magnetic resonance imaging. Relative cerebral blood flow change maps were calculated for comparison. RESULTS: The spatial relation between the sensorimotor cortex and the lesion could be determined in all cases. Additional activations covered the ipsilateral sensorimotor cortex and the bilateral cerebellum, premotor cortices and supplementary motor areas. Patients with motor symptoms of the stimulated hand (paresis, focal seizures) activated the ipsilateral premotor cortices and contralateral cerebellum more often than patients without motor symptoms. The SPM results for p<0.005 and cerebral blood flow change maps showed considerably overlapping motor area activations. For p<0.001, SPM missed three sensorimotor cortex activations depicted by cerebral blood flow change maps and by SPM for p<0.005 in typical localisation. SPM analyses showed less activations probably unrelated to task performance. CONCLUSION: It is concluded that SPM provides an efficient method for analysing individual preoperative PET activation studies. Activations of the ipsilateral premotor cortices and contralateral cerebellum may indicate an enhanced recruitment of ipsilateral motor pathways evoked by functional reorganisation processes. However, this changed activation pattern was not necessarily associated with a better neurological status. PMID- 12640067 TI - Is a combination of Tc-SPECT or perfusion weighted magnetic resonance imaging with spinal tap test helpful in the diagnosis of normal pressure hydrocephalus? AB - OBJECTIVE: The aim of this study was to evaluate the combination of spinal tap test (STT) with cerebral perfusion measurement assessed either by Tc-bicisate SPECT (Tc-SPECT) or perfusion weighted MRI (pwMRI), or both, for a better preoperative selection of promising candidates for shunt operations in suspected idiopathic normal pressure hydrocephalus. METHODS: 27 consecutive patients were examined with a standard clinical protocol (assessed by the Homburg Hydrocephalus Scale (HHS)) as well as with 99m Tc-bicisate-SPECT (n=27) or additionally by pwMRI (n=12) before and after STT. The results of these examinations were compared preoperatively for each patient and correlated with postoperative clinical outcome after shunt surgery. RESULTS: Nine patients showed both, a clinical improvement, and increased cerebral perfusion after STT. They underwent shunt surgery with good to excellent results. In another nine patients increasing cerebral perfusion was detected although they did not show a clear clinical improvement after STT. Six of them also received a shunt operation with good to excellent outcome. Three patients of the last group could have an operation. Nine patients did not show any clinical improvement or any kind of increasing cerebral perfusion after STT. Therefore, they did not undergo surgery. The results of SPECT and pwMRI correlated in 92 % of the patients (11 of 12). CONCLUSION: It is concluded that a combination of clinical assessment with SPECT or pwMRI is helpful in the preoperative selection of patients for shunting procedures with suspected NPH syndrome. This combination is a minimal invasive and objective test modality that is superior to STT alone. Further studies are necessary for a comparison of the described imaging techniques with different diagnostic tests in this difficult field of cerebral disease. PMID- 12640069 TI - Treatment of IgM antibody associated polyneuropathies using rituximab. AB - OBJECTIVES: Polyneuropathies with associated serum IgM antibodies are often difficult to treat. Rituximab is a monoclonal antibody directed against the B cell surface membrane marker CD20. Rituximab eliminates B cells from the circulation, and, over time, could reduce cells producing autoantibodies. This study tested the ability of rituximab to produce changes in serum antibody titres, and improvement in strength, in patients with neuromuscular disorders and IgM autoantibodies. METHODS: Over a period of two years, the authors evaluated changes in strength, measured by quantitative dynamometry, and concentrations of several types of serum antibodies in patients with polyneuropathies and serum IgM autoantibodies. Twenty one patients treated with rituximab were compared with 13 untreated controls. RESULTS: Treatment with rituximab was followed by improved strength (an increase of mean (SEM) 23% (2%)of normal levels of strength), a reduction in serum IgM autoantibodies (to 43% (4%) of initial values), and a reduction in total levels of IgM (to 55% (4%) of initial values). There was no change in levels of serum IgG antibodies. There were no major side effects, even though B cells were virtually eliminated from the circulation for periods up to two years. CONCLUSIONS: In patients with IgM autoantibody associated peripheral neuropathies, rituximab treatment is followed by reduced serum concentrations of IgM, but not IgG, antibodies, and by improvement in strength. Additional studies, with placebo controls and blinded outcome measures, are warranted to further test the efficacy of rituximab treatment of IgM associated polyneuropathies. PMID- 12640071 TI - Beta endorphin concentrations in PBMC of patients with different clinical phenotypes of multiple sclerosis. AB - The possible link between the opioid peptide beta endorphin and the heterogeneity of the clinical course of multiple sclerosis (MS) was investigated. Peripheral blood mononuclear cells (PBMC) concentrations of beta endorphin were measured in 50 patients in different phases of MS. Thirty nine patients also underwent post contrast magnetic resonance imaging of the brain. Among MS forms, the highest beta endorphin concentrations were found in PBMC from patients with relapsing remitting MS and the lowest in patients with the progressive forms. Average beta endorphin concentrations were lower, although not significantly, in patients with than in those without magnetic resonance imaging enhanced lesions. These data suggest that beta endorphin may have a role in the downregulation of the inflammatory process. PMID- 12640070 TI - Neuromuscular and sensory disturbances in patients with well treated coeliac disease. AB - OBJECTIVES: A body of evidence shows that coeliac disease is associated with protean manifestations outside the intestine, and neurological disorders are well recognised. However, it remains obscure whether there are signs of clinical or subclinical nervous system involvement even in patients adopting an adequate gluten free diet. The aim of this study was to assess in a controlled study whether patients with treated coeliac disease carry an increased risk for neuropathy and characterise the type of possible neuropathy. METHODS: Electroneuromyographic findings and vibration, thermal, and tactile thresholds of 26 patients with coeliac disease and 23 patients with reflux disease were analysed. RESULTS: Six (23.1 %) coeliac disease patients and one (4.3 %) reflux disease patient showed findings of chronic axonal neuropathy in quantitative needle EMG. In addition, two coeliac disease patients showed findings suggestive for myopathy. There were no significant differences in warm, cold, or vibration thresholds between the groups but means of heat pain thresholds and tactile thresholds were significantly higher in coeliac patients than in controls. CONCLUSION: An increased occurrence of axonal neuropathy was observed in well treated coeliac disease. This further indicates that neurological manifestations occur even in patients without overt malabsorption. PMID- 12640072 TI - Motor evoked potentials from the pelvic floor in patients with multiple sclerosis. AB - The use of motor evoked potentials (MEPs) to study the integrity of pelvic floor motor innervation is poorly described in the literature. This study evaluated the clinical use of pelvic floor MEPs in 16 women with multiple sclerosis. Lower urinary tract dysfunction was assessed with urodynamic investigations. Transcutaneous magnetic stimulation was applied over the motor cortex and spinal roots, and MEPs were recorded from the puborectalis, the external urethral sphincter, and the abductor hallucis muscles. In many patients, responses from the pelvic floor muscles could not be evoked, and central motor conduction times for the puborectalis motor pathways could only be calculated in 56%. There was a poor correlation of abnormal conduction to lower urinary tract dysfunction. It is concluded, that unevokable responses from pelvic floor muscles in a patient with multiple sclerosis should be interpreted with care, and that pelvic floor MEPs have a limited clinical value in the investigation of suspected demyelinating disease. PMID- 12640073 TI - A diffusion tensor magnetic resonance imaging study of brain tissue from patients with migraine. AB - OBJECTIVE: To measure in vivo, using diffusion tensor magnetic resonance imaging (DT-MRI) the extent of pathological damage of normal appearing brain tissue (NABT) from patients with migraine. METHODS: Dual echo and DT-MRI scans of the brain were acquired from 34 patients with migraine and 17 sex and age matched healthy volunteers. Mean diffusivity (MD) and fractional anisotropy (FA) histograms of the NABT were obtained from all subjects and the histograms' peak heights and average NABT MD and FA measured. When present, average MD and FA values of T2 visible lesions were also measured. RESULTS: In comparison with healthy volunteers, patients with migraine had lower MD histogram peak height (p=0.02) of the NABT. No differences were found in FA histogram derived metrics between migraine patients and healthy subjects. No difference was found for any MD and FA histogram derived metrics between migraine patients with and without brain MRI lesions, and between patients with and without aura. CONCLUSIONS: This study shows that, although brain damage may extend beyond T2 weighted abnormalities in patients with migraine, the severity of these "occult" changes is mild compared with that found in other diseases associated with white matter abnormality. PMID- 12640075 TI - Paraneoplastic ophthalmoplegia and subacute motor axonal neuropathy associated with anti-GQ1b antibodies in a patient with malignant melanoma. AB - A 68 year old woman developed oculomotor paresis shortly after metastatic progression of her melanoma was discovered. She was then immunised with the tumour antigen MAGE-3 in combination with an immunological adjuvant. During immunisation her symptoms worsened and she developed severe, predominantly proximal axonal motor neuropathy and became bedridden. IgM antibodies against gangliosides GM2, GD3, and GQ1b were detected in serum obtained two weeks before and nine weeks after the onset of symptoms. Immunohistochemically, the patient's IgM reacted with the tumour and co-localised with GQ1b. She improved neurologically following steroid treatment and became ambulatory. PMID- 12640074 TI - Thyrotoxic autoimmune encephalopathy: a repeat positron emission tomography study. AB - Thyroid related autoantibodies have been related to the development of encephalopathy, known as Hashimoto's encephalopathy. However, their relation with the encephalopathy occurring in patients with Graves' disease has not been well established. The case is reported of a 51 year old woman presenting with subacute progressive dementia with evidence of hyperthyroidism. She had Graves' disease associated with high titres of thyroid related autoantibodies. Her encephalopathy was not improved by antithyroid drugs, but promptly responded to corticosteroid treatment, and stabilised with a gradual reduction of thyroid related autoantibody titres. Brain positron emission tomography initially showed a diffuse and multifocal cerebral hypometabolism with subsequent normalisation on her clinical recovery, which was consistent with the acute and reversible cerebral inflammation probably mediated by autoimmune mechanisms. PMID- 12640076 TI - Middle cerebral artery territory infarction sparing the precentral gyrus: report of three cases. AB - We report three patients with large middle cerebral artery infarctions in the non dominant hemisphere, with striking recovery of motor function. In each case this excellent functional outcome correlated with selective sparing of the motor cortex in the precentral gyrus. We discuss some of the possible circulatory variants that might underlie this pattern of infarction. PMID- 12640077 TI - Effects of decompressive craniectomy on brain tissue oxygen in patients with intracranial hypertension. AB - This report examined the intraoperative course of partial pressure of brain tissue oxygen (P(ti)O(2)) and intracranial pressure (ICP) during surgical decompressive craniectomy for medically intractable intracranial hypertension due to diffuse brain swelling in three patients after severe subarachnoid haemorrhage and aneurysm coiling. The mean ICP decreased from 59 mm Hg to 10 mm Hg in a two step fashion, relating to bone flap removal and dural opening. Simultaneously, P(ti)O(2) increased rapidly from 0.8 kPa (6 mm Hg) to 3.07 kPa (23 mm Hg). P(ti)O(2) and ICP remained at non-critical ranges postoperatively. Despite these beneficial effects on ICP and P(ti)O(2), the patients' clinical status remained poor with two in a persistent vegetative state and one dead. PMID- 12640078 TI - Fridtjof Nansen (1861-1930). PMID- 12640079 TI - American trypanosomiasis (Chagas' disease): an unrecognised cause of stroke. AB - BACKGROUND: American trypanosomiasis, known as Chagas' disease (CD) is a major cause of cardiomyopathy in South America. Irreversible damage to the heart can appear 10 to 20 years after chagasic infection. The frequency of cerebrovascular complications in chronic CD is unknown. OBJECTIVES: To describe a group of patients with chronic or latent CD affected by ischaemic stroke and identify predictive variables for stroke in CD patients. PATIENTS AND METHODS: Retrospective case series of stroke patients with CD was studied using a cross sectional, descriptive design. CD was confirmed by positive immunofluorescence and haemaglutination serology. Data were collected on age, sex, vascular risk factors, previous history of CD, diagnostic stroke subtype, electrocardiograph and echocardiography findings. Frequency of vascular risk factors were compared with a control group of 239 non-chagasic stroke patients. RESULTS: 136 consecutive CD stroke patients, mean age 56 years, 72 women and 64 men were identified. Vascular risk factors were observed in 81.6% of CD patients. Hypertension (70.29% versus 51.47%; p=0.0004), diabetes mellitus (15.9% versus 6.61%; p=0.0143), and tobacco use (53.98% versus 30.88%; p=0.00002) were significantly less frequent in the CD stroke group. Cardiomyopathy was significantly higher in CD stroke patients (45.58% versus 24.69%; p=0.00005). Abnormal electrocardiograms was observed in 82% of chagasic patients (right bundle branch block 39.5%, left anterior fascicular block 35.8%). Left ventricle (LV) diastolic dysfunction (61.47%), LV systolic dysfunction (51.18%), congestive cardiomyopathy (29.92%), and apical aneurysm (15.74%) were the most frequent echocardiographic findings. Aetiologies were cardioembolism (52.2%), undetermined (36.76%), atherothrombotic (8.82%), and small vessel stroke (2.2%). A diagnosis of CD was established after presentation with stroke in 38.23% of the patients. CONCLUSIONS: CD should be included in the differential diagnosis of stroke in patients of South American origin. PMID- 12640080 TI - Surgery for Parkinson's disease: lack of reliable clinical trial evidence. AB - There has been a striking resurgence of interest in surgery for Parkinson's disease (PD) with new targets identified and new procedures developed. This systematic review identified over 500 studies of surgery for PD published since 1990, including over 10 000 patients. However, the authors were unable to assess the value of PD surgery reliably because only seven randomised trials were identified including just 196 patients. Studies of surgery for PD have generally been of poor quality with too few patients, too short follow up, inappropriate choice of outcome measures, and lack of control groups. Much larger, randomised, controlled trials are needed to assess the longer term effects of surgery on patient rated quality of life and cost effectiveness. PMID- 12640081 TI - High concentrations of procollagen propeptides in chronic subdural haematoma and effusion. AB - Procollagen propeptides increase in the CSF after subarachnoid haemorrhage, reflecting increased collagen synthesis in the arachnoid. We studied the induction of dural collagen synthesis after cerebral trauma by measuring the carboxyterminal propeptide of type I procollagen (PICP) and the aminoterminal propeptide of type III procollagen (PIIINP) in 17 subdural haematoma or effusion fluid samples obtained at operation on days 10-85 after head trauma. The concentration of PICP was 78-fold higher and that of PIIINP 156-fold higher, relative to that in the serum. These results indicate that meningeal trauma is followed by a long lasting increase in dural collagen synthesis, and suggest that enhanced synthesis of the various extracellular matrix components may have a role in the development of chronic subdural haematoma or effusion. PMID- 12640083 TI - Adult onset Niemann-Pick disease type C presenting with psychosis. AB - Niemann-Pick disease type C (NPC) is an autosomal recessive neurometabolic disorder that rarely presents in adulthood, and is associated with cognitive decline, various movement disorders (ataxia, chorea, dystonia, and myoclonus), a vertical supranuclear gaze palsy (VSGP), and seizures. A recent case report demonstrated a delay in diagnosis of eight years when a patient with NPC presented with psychosis. This article reviewed all cases seen at the Mayo Clinic with a possible diagnosis of NPC between 1976 and 2000. Of the 52 possible cases, five had an established diagnosis of adult onset NPC. Of these, two presented with psychosis and were not diagnosed with NPC for 5 and 15 years, respectively. NPC may initially present in adulthood with psychosis, and when psychosis is associated with VSGP, various dyskinesias, and seizures, NPC should be suspected. PMID- 12640082 TI - Ictal magnetoencephalographic discharges from elementary visual hallucinations of status epilepticus. AB - PURPOSE: To report the rare opportunity to study ictal magnetoencephalography (MEG) in a 26 year old man with simple partial status epilepticus that presented as elementary visual hallucinations (EVHs) in the right upper visual field. METHODS: The patient described his EVHs as "snowing on TV," "flickering lights," and "rotating coloured balls" that continued for several days. MEG and simultaneous EEG were recorded twice: during an episode of EVHs (ictal recordings) and after EVHs were controlled by medications (interictal recordings). RESULTS: During EVHs, MEG showed continuous periodic epileptiform discharges over the left posterior superior temporal region, while simultaneous EEG showed rhythmic theta waves and sporadic spikes over the left temporal region. The MEG discharge consisted of a three phase spike complex. Equivalent current dipoles (ECDs), modelled from spike complexes, localised in the left superior temporal area. After drug treatment controlled the EVHs, interictal MEG and EEG showed rare spikes over the same left temporal region. The average ictal ECD moment (mean (SD)) (128.7 (32.8 nAm)) was significantly weaker than the average interictal ECD moment (233.0 (63.9) nAm) (p<0.05). CONCLUSIONS: The continuous, periodic, and clustered discharges seen on ictal MEG were the sources of EVH. The weaker ictal ECD sources were frequently not detected by scalp EEG, while the stronger interictal sources, presumably originating from an extensive interictal zone, were sufficiently large to be seen as EEG spikes. PMID- 12640085 TI - Measurement of pulse pressure profiles in patients with trigeminal neuralgia. AB - Applanation tonometry is a non-invasive method of assessing the arterial blood pressure profiles in both the peripheral and systemic circulation. In this study the authors examined whether there were differences in these profiles in patients with trigeminal neuralgia. The carotid artery and derived aortic blood pressure waveforms were obtained using a pulse wave analysis system. The ratio of the pressure wave amplitude above the systolic shoulder to the total systolic blood pressure (augmentation index, AIx) was recorded. Thirty two patients with trigeminal neuralgia (16 male and 16 female) and 100 controls (50 male and 50 female) were recruited. Eleven patients had been treated by microvascular decompression, mean (SD) time from surgery 17 (24) months (range 3-86 months). For the patients with trigeminal neuralgia, the right and left carotid artery AIxs (mean (SD)) were 120.6 (21.7)% and 120.7 (19.1)% respectively. Corresponding values for the control group were 120.5 (19.3)% and 120.9 (19.5)%. The calculated AIx for the ascending aorta was 27.7 (10.1)% and 27.2 (10.5)% for the patients with trigeminal neuralgia and controls respectively. No significant differences were seen in either the right or left carotid artery (p=0.5 and p=0.6 respectively) or the derived ascending aorta (p=0.8). The results show that there does not seem to be a generalised increase in arterial stiffness in patients with trigeminal neuralgia. PMID- 12640084 TI - Interlocking finger test: a bedside screen for parietal lobe dysfunction. AB - This study sought to determine the utility of an interlocking finger task in screening for parietal lobe dysfunction. The ability of 69 patients to imitate a standardised set of four interlocking finger figures was compared with concurrent performance on formal neurocognitive tests. Poor interlocking finger test scores correlated most highly with standard measures of parietal lobe dysfunction. In addition, an analytical model of parietal dysfunction indicated the interlocking finger test was similar to, if not better than, standard tests of parietal lobe dysfunction. Attempts to imitate these figures should serve as a fast and simple screen of parietal lobe dysfunction. PMID- 12640086 TI - Brain cysts associated with mutation in the Aristaless related homeobox gene, ARX. AB - The novel Aristaless related homeobox gene, ARX, is widely expressed in the brain and is thought to play a key role in the regulation of brain development. Neurological phenotypes caused by ARX mutations have recently started to unfold. We describe a 72 year old man with X-linked mental retardation due to a 24 bp duplication mutation in exon 2 of the ARX gene. Cerebral MRI showed bilateral cystic-like cavities in both the cerebral and cerebellar hemispheres. No retraction or expansion in neighbouring parenchyma was observed, there was no history of acute neurological impairment, and no risk factors for cerebrovascular disease were found. The lesions appeared to be congenital and represented benign developmental cysts, possibly caused by the ARX mutation. PMID- 12640087 TI - Sudden bilateral simultaneous deafness with vertigo as a sole manifestation of vertebrobasilar insufficiency. AB - A 68 year old woman presented with bilateral sudden simultaneous hearing loss and transient spontaneous vertigo as a sole manifestation of vertebrobasilar insufficiency. Extensive investigation to exclude other causes was unremarkable. Magnetic resonance imaging of the brain, including diffusion images, showed no abnormalities. A magnetic resonance angiogram showed severe stenosis of the middle third of the basilar artery. A pure tone audiogram showed moderate sensorineural-type hearing loss bilaterally. The localisation and mechanism of an isolated cochleovestibular dysfunction are discussed. PMID- 12640088 TI - The natural course of Sneddon syndrome: clinical and magnetic resonance imaging findings in a prospective six year observation study. AB - Sneddon syndrome (SS) is increasingly recognised as a cause of ischaemic stroke in young adults. As the natural course of SS is not well defined, the authors performed a prospective six year clinical and neuroradiological follow up study. Thirteen patients with definite diagnosis of SS (livedo racemosa, characteristic skin biopsy, and history of stroke) entered a follow up programme that consisted of clinical examinations, two magnetic resonance imaging (MRI) investigations, and a comprehensive laboratory follow up protocol. The most frequent clinical findings during follow up had been headache (62%) and vertigo (54%). Seven patients (54%) suffered from transient ischaemic attacks, however, completed stroke has not been obtained during follow up. Progression of white matter lesions detected in MRI were present in 10 of 13 patients. Laboratory follow up protocol revealed transient antiphospholipid antibodies in two subjects. This prospective six year follow up study suggests a low incidence of territorial stroke but outlines progressive leucencephalopathy in patients with SS. PMID- 12640089 TI - Effect of deprivation on time to hospital in acute stroke. PMID- 12640090 TI - Paraoxonase 1 promoter and coding region polymorphisms in Parkinson's disease. PMID- 12640091 TI - Immunohistochemical study of caveolin-3 in idiopathic hyperCKaemia. PMID- 12640092 TI - Multifocal myoclonus secondary to tranexamic acid. PMID- 12640093 TI - Dysphagia due to Chiari I malformation mimicking ALS. PMID- 12640094 TI - Benefit of folic acid supplementation in parkinsonian patients treated with levodopa. PMID- 12640095 TI - High dose intravenous immune globulin in the treatment of hereditary recurrent brachial plexus neuropathy. PMID- 12640097 TI - Value of mesothelin immunostaining in the diagnosis of mesothelioma. AB - Mesothelin is a cell surface antigen of unknown function that is strongly expressed in mesothelial cells. Although it was reported in 1992 that immunostaining with the K1 anti-mesothelin antibody could be very useful in distinguishing between epithelioid mesotheliomas and pulmonary adenocarcinomas, no further studies have been published on the value of this marker in the diagnosis of mesotheliomas. To determine whether mesothelin can assist in discriminating epithelioid mesotheliomas from lung adenocarcinomas or from other carcinomas metastatic to the serosal membranes, 55 mesotheliomas (44 epithelioid, 3 biphasic, and 8 sarcomatoid), 48 carcinomas of the lung (31 adenocarcinomas, 17 squamous carcinomas), and 86 nonpulmonary adenocarcinomas (14 ovary, 5 peritoneum, 9 endometrium, 11 pancreas, 4 stomach, 16 colon, 12 breast, 9 kidney, 4 thyroid, and 2 prostate) were investigated for mesothelin expression using the recently available 5B2 anti-mesothelin monoclonal antibody. Reactivity was obtained in all 44 (100%) of the epithelioid mesotheliomas, 12 (39%) of the lung adenocarcinomas, and 42 (49%) of the nonpulmonary adenocarcinomas (14 [100%] ovary; 5 [100%] peritoneum; 6 [67%] endometrium; 10 [91%] pancreas; 2 [50%] stomach; 5 [31%] colon; and in none [0] of the breast, kidney, thyroid, or prostate). Three (18%) of the squamous carcinomas of the lung, but none of the sarcomatoid mesotheliomas, exhibited positivity for this marker, nor was any reactivity seen in the spindle cell component of the biphasic mesotheliomas. It is concluded that despite the low specificity of mesothelin for discriminating between epithelioid mesotheliomas and adenocarcinomas, immunostaining for this marker may have some utility in those instances in which the results obtained with the standard panel of immunohistochemical markers used for the diagnosis of mesotheliomas are equivocal. Because mesothelin is a highly sensitive positive marker for epithelioid mesotheliomas, a negative staining for this marker is an indication against such a diagnosis; however, because of its limited utility, it is not recommended for inclusion in the standard panel of immunohistochemical markers used in the distinction between mesotheliomas and adenocarcinomas. PMID- 12640096 TI - Immunohistochemical expression of CK20, p53, and Ki-67 as objective markers of urothelial dysplasia. AB - Urothelial dysplasia and carcinoma in situ (CIS) are related to recurrence and progression of urothelial carcinoma. Distinguishing CIS and dysplasia from reactive atypia is often difficult on the basis of histological features alone. Cytokeratin 20 (CK20), p53, and Ki-67 are related either to neoplastic change or prognosis in urothelial proliferations. The objective of the present study was to establish the immunohistochemical pattern of these three antibodies in urothelial dysplasia and CIS. Three groups of patients were evaluated: 40 nonneoplastic urothelial samples, 50 cases with histologically incontrovertible CIS, and 30 samples with nonconclusive atypical changes (atypia of unknown significance). Monoclonal antibodies (MoAb) against CK20, p53, and Ki-67 (MIB-1) were used on paraffin-embedded samples. Nonneoplastic urothelium showed no reactivity to CK20 except for umbrella cells; p53 and Ki-67 were negative or weakly positive in <10% of basal cells. In the CIS group, 42% showed positivity for all three MoAb; 44%, for two; and 14%, only for one. CK20 was positive through the full thickness of the urothelium in 72% of cases, p53 was positive in 80% of cases, and Ki-67, in 94% of cases. In the third group, the suspected dysplastic cells showed strong positivity in scattered cells through the epithelium in 75% of cases. Aberrant CK20 expression in urothelial cells plus overexpression of p53 and Ki-67 are indicators of dysplastic change in urothelial mucosa. Thus, immunohistochemistry is a useful tool to confirm the diagnosis of CIS and could be helpful to distinguish dysplastic changes from reactive atypia. PMID- 12640098 TI - Prognostic significance of matrix metalloproteinase 2 and tissue inhibitor of metalloproteinase 2 expression in prostate cancer. AB - Matrix metalloproteinases (MMPs) are proteolytic enzymes capable of degrading the structural support network for normal and malignant cells, promoting neoplastic cell invasion and metastasis. Tissue inhibitors of metalloproteinases (TIMPs) maintain connective tissue integrity by modulating MMP activity. Formalin-fixed paraffin-embedded tissue sections from 138 prostatic adenocarcinomas (PACs) were immunostained by a combined automated/manual method using monoclonal antibodies against MMP2 and TIMP2. Immunoreactivity was semiquantitatively scored based on stain intensity and distribution, and results were correlated with Gleason grade, pathologic stage, ploidy status, and disease recurrence. One hundred five of 138 (76%) and 113/138 (82%) PACs expressed MMP2 and TIMP2, respectively. Co expression was observed in 94/138 (68%) of PACs (P =.01), correlated with advanced tumor stage (P =.05), and tended to be associated with disease recurrent cases (P =.07). TIMP2 expression individually correlated with advanced tumor stage (P =.04) and reached near significance with disease recurrence (P =.06). MMP2 expression was also more frequent in recurrent PACs, although this value did not reach statistical significance (P =.07). However, on multivariate analysis, only pathologic stage (P =.009) and ploidy status (P =.03) independently predicted disease recurrence. In conclusion, MMP2 and TIMP2 are co-expressed in a majority of PACs and correlate with prognostic variables. Interestingly, contrary to the previously documented anti-tumor effects of TIMPs, TIMP2 expression appears to have a tumor-promoting role in PACs and warrants further investigation. PMID- 12640099 TI - Assessment of utility of ductal lavage and ductoscopy in breast cancer-a retrospective analysis of mastectomy specimens. AB - Early detection of breast lesions continues to be an important goal in the management of breast cancer. At present, mammographic imaging in addition to physical examination is the main screening method for the detection of cancer. Fiberoptic ductoscopy and duct lavage are being recently used to evaluate patients at risk for breast cancer. Both techniques examine the nipple and central duct area to identify intraductal lesions. In this study, we examined the frequency of involvement of these structures in mastectomy specimens as a surrogate marker to estimate the utility of these methods in breast cancer patients. The presence and type of involvement of the nipple and central duct area was retrospectively evaluated in 801 mastectomy specimens from a 4-year period that had been performed for infiltrating or in situ carcinoma. Atypical proliferation or cells, when seen in the ducts of this region, was considered as evidence of nipple involvement, even if definite evidence of malignancy was lacking. The review of 801 mastectomies showed nipple and central duct involvement in 179 (22%) cases. Among the 665 cases of infiltrating carcinoma, 17% did not have an intraductal component. The relative rarity of nipple and central duct in mastectomy specimens and the lack of an in situ component in many cases raise questions about the utility of fiberoptic ductoscopy and duct lavage as methods for screening of breast cancer. Additionally, as these methods examine only 1-2 ducts of the 15-20 ducts that open at the nipple, they might fail to detect focal abnormalities. PMID- 12640100 TI - Cyclin alterations in giant cell tumor of bone. AB - Cyclins play an important role in regulating the passage of dividing cells through critical checkpoints in the cell cycle. Because alterations of several cyclins, especially cyclin D1, have been implicated in the development of many human neoplasms, we examined 32 cases of giant cell tumor of long bones for cyclin D1 gene amplification and protein overexpression using differential polymerase chain reaction and immunohistochemistry, respectively. In addition, the expression of cyclin D3, cyclin B1, and the proliferation-associated antigen Ki-67 (MIB-1) was assessed immunohistochemically. Low-level cyclin D1 gene amplification was detected in 61% of giant cell tumor cases. All tumors showed cyclin D1, cyclin D3, cyclin B1, and Ki-67 (MIB-1) staining; however, the distribution was very characteristic. Cyclin D1 protein expression was seen predominantly in the nuclei of the giant cells, with occasional mononuclear cells staining. There was no correlation between cyclin D1 gene amplification and protein overexpression. Cyclin D3 staining showed a similar distribution, with 88% of cases showing protein overexpression. Cyclin D1 and/or D3 staining in the giant cells was never associated with staining for either cyclin B1 or Ki-67 (MIB 1), as the expression of the latter two proteins was restricted to the mononuclear cells. Cyclin B1 overexpression was seen in 44% of cases. Ki-67 (MIB 1) staining was present in all cases, and between 10 to 50% of the mononuclear cells were positive. These results suggest that alterations in cyclin D1 and/or D3 might play a role in the pathogenesis of giant cell tumor of bone. PMID- 12640101 TI - Calcifications in ovary and endometrium and their neoplasms. AB - In this study, we investigated the role of hormones in the pathogenesis of calcifications in ovary and in endometrium and their neoplasms of the gynecologic tract and assessed the anatomic location and incidence of these calcifications. The study consists of three parts designed to investigate the pathogenesis, the location, and the incidence of calcifications in ovary and endometrium and their neoplasms. In the first part, 79 female guinea pigs were divided into 10 groups, and different hormones, given weekly for 12 months, were administered to the guinea pigs by group. A control group of 7 guinea pigs received sterile water. Calcifications developed in 5 of 7 guinea pigs treated with prolactin, 10 of 20 treated with human chorionic gonadotropin, 5 of 11 treated with estradiol, 3 of 7 treated with estrone, 1 of 6 treated with growth hormone, and 1 of 10 treated with testosterone; in 20 of the guinea pigs, the calcifications developed in the stroma of the endometrium, and in 5 guinea pigs, they developed in the ovary. The second part of the study consisted of an evaluation of the specific location of calcifications in 43 consecutive human surgical ovaries and endometria. Calcifications were seen only in the stroma in 100% of the ovarian serous adenofibroma specimens; in ovarian serous borderline neoplasms, the stroma contained 70 to 100% of the calcifications, and the epithelium had 0 to 30% of the calcifications. In ovarian serous carcinoma specimens, the calcifications were seen in the stroma in 50 to 60% of the cases, in the epithelium in 40% of the cases, and in areas of necrosis in 10% of the cases. The third part of the study was directed to determine the frequency of calcifications in ovarian lesions. We found that all cases of endosalpingiosis and ovarian low-grade serous carcinoma had calcifications, whereas 80% of the cases of serous borderline tumor had calcifications, and only 50% of the cases of ovarian high-grade serous carcinoma contained calcifications. The results of this study indicate that the majority of the calcifications in the ovary and the endometrium and their neoplasms are present in the stroma. This is most probably secondary to metabolic changes, which could be related to hormones and not caused by degenerative changes in epithelial cells. PMID- 12640102 TI - Lymphocytic mastitis and diabetic mastopathy: a molecular, immunophenotypic, and clinicopathologic evaluation of 11 cases. AB - Lymphocytic mastitis and diabetic mastopathy are uncommon fibroinflammatory breast diseases. The lesions seen in these entities are unique in that the associated lymphoid infiltrates are composed of predominantly B cells. In addition, B-cell lymphoepithelial lesions, a finding commonly associated with extranodal marginal zone B-cell/mucosa-associated lymphoid tissue (MALT) lymphomas, are also often present in lymphocytic mastitis and diabetic mastopathy. Although the clinical and immunomorphologic features are well characterized, the clonality of the B-cell infiltrate and the lymphomatous potential of lymphocytic mastitis and diabetic mastopathy have not been emphasized in the literature. We evaluated 11 cases of lymphocytic mastitis/diabetic mastopathy for immunoglobulin heavy chain gene rearrangement and correlated the findings with all available clinical data. A longstanding history of Type I diabetes mellitus was present in seven patients. One nondiabetic patient had Sjogren's syndrome, and two patients had no history of diabetes mellitus or other autoimmune disease. Clinical data were unavailable for one patient. B-cell-predominant lymphoid infiltrates were seen in all cases, and B-cell lymphoepithelial lesions were found in five. No evidence of a B-cell clone was found in any of the 11 cases by appropriately controlled immunoglobulin heavy chain gene rearrangement studies, and none of the patients developed lymphoma during follow-up intervals ranging from 2-126 months. These findings suggest that despite the presence of B-cell-predominant lymphoid infiltrates and lymphoepithelial lesions, lymphocytic mastitis and diabetic mastopathy do not appear to be associated with an increased risk for lymphoma. PMID- 12640103 TI - Immunophenotype of desmoplastic small round cell tumors as detected in cases with EWS-WT1 gene fusion product. AB - Desmoplastic small round cell tumor is a rare tumor typically involving peritoneum. Although the histogenesis of desmoplastic small round cell tumor has yet to be elucidated, immunophenotypical and morphological analysis shows a characteristic divergent phenotype overlapping with other round cell tumors such as Ewing's sarcoma/primitive neuroectodermal tumor, rhabdomyosarcoma, small cell mesothelioma, and carcinoma. Detection of the EWS-WT1 gene fusion is characteristic of desmoplastic small round cell tumor and has been used reliably in tumor diagnosis. In this study, we evaluated the immunophenotype of 23 desmoplastic small round cell tumor cases with the EWS-WT1 gene fusion product identified by reverse transcription-polymerase chain reaction. Paraffin sections were stained with antibodies against calretinin, WT1 (C19), desmin, myoglobin, MyoD, Myf5, myogenin, placental alkaline phosphatase, cytokeratins, MIC2, HER2/neu and c-kit using standard immunohistochemical methods. Immunoreactivity was evaluated semiquantitively by light microscopy. Desmoplastic small round cell tumors showed reactivity with calretinin in 4/21, desmin in 21/23, myoglobin in 5/17, placental alkaline phosphatase in 17/21, HER2/neu in 7/18 (3+ in 1 and 1+ in 6), c-kit in 2/14, MIC2 in 13/23, WT1 in 16/23, CAM5.2 in 21/23, and AE1/3 in 16/23 cases. The most sensitive myogenic and epithelial markers are desmin and CAM 5.2. Although nuclear reactivity of the early myogenic regulatory factors (MyoD, myogenin, Myf5) was not detected, myoglobin immunoreactivity was present in 29% of desmoplastic small round cell tumors. HER2/neu overexpression (3+) and c-kit expression are uncommon in desmoplastic small round cell tumors. A panel of myogenic and epithelial markers should be used to detect the divergent phenotype in desmoplastic small round cell tumors, a key feature in the differential diagnosis. Detection of EWS-WT1 fusion becomes critical for the diagnosis when the characteristic divergent phenotype cannot be detected immunohistochemically. PMID- 12640104 TI - Primary ear and temporal bone meningiomas: a clinicopathologic study of 36 cases with a review of the literature. AB - "Primary" ear and temporal bone meningiomas are tumors that are frequently misdiagnosed and unrecognized, resulting in inappropriate clinical management. To date, a large clinicopathologic study of meningiomas in this anatomic site has not been reported. Thirty-six cases of ear and temporal bone meningiomas diagnosed between 1970 and 1996 were retrieved from our files. Histologic features were reviewed, immunohistochemical analysis was performed (n = 19), and patient follow-up was obtained (n = 35). The patients included 24 females and 12 males, aged 10-80 years (mean, 49.6 years), with female patients presenting at an older age (mean, 52.0 years) than male patients (mean, 44.8 years). Patients presented clinically with hearing changes (n = 20), otitis (n = 7), pain (n = 5), and/or dizziness/vertigo (n = 3). Symptoms were present for an average of 24.6 months. The tumors affected the middle ear (n = 25), external auditory canal (n = 4), or a combination of temporal bone and middle ear (n = 7). The tumors ranged in size from 0.5 to 4.5 cm in greatest dimension (mean, 1.2 cm). Radiographic studies demonstrated a central nervous system connection in 2 patients. Histologically, the tumors demonstrated features similar to those of intracranial meningiomas, including meningothelial (n = 33), psammomatous (n = 2), and atypical (n = 1). An associated cholesteatoma was identified in 9 cases. Immunohistochemical studies confirmed the diagnosis of meningioma with positive reactions for epithelial membrane antigen (79%) and vimentin (100%). The differential diagnosis includes paraganglioma, schwannoma, carcinoma, melanoma, and middle ear adenoma. Surgical excision was used in all patients. Ten patients developed a recurrence from 5 months to 2 years later. Five patients died with recurrent disease (mean, 3.5 years), and the remaining 30 patients were alive (n = 25, mean: 19.0 years) or had died (n = 5, mean: 9.5 years) of unrelated causes without evidence of disease. We conclude that extracranial ear and temporal bone meningiomas are rare tumors histologically similar to their intracranial counterparts. They behave as slow-growing neoplasms with a good overall prognosis (raw 5-y survival, 83%). Extent of surgical excision is probably the most important factor in determining outlook because recurrences develop in 28% of cases. PMID- 12640106 TI - Most malignant fibrous histiocytomas developed in the retroperitoneum are dedifferentiated liposarcomas: a review of 25 cases initially diagnosed as malignant fibrous histiocytoma. AB - Forty-four samples from 25 cases of retroperitoneal sarcoma initially diagnosed as malignant fibrous histiocytoma were histologically reviewed. Immunohistochemistry for mdm2 and cdk4 was performed on 20 cases. Comparative genomic hybridization was performed on 18 samples from 13 patients. Seventeen cases were reclassified as dedifferentiated liposarcoma. Twenty-one of 32 samples from these patients showed areas of well-differentiated liposarcoma, allowing the diagnosis of dedifferentiated liposarcoma. Immunohistochemistry performed in 15 of these cases showed positivity for mdm2 and cdk4. Comparative genomic hybridization analysis performed on 15 samples from 11 of these patients showed an amplification of the 12q13-15 region. Eight cases were reclassified as poorly differentiated sarcoma. Twelve samples from these patients showed no area of well differentiated liposarcoma. Immunohistochemistry showed positivity for mdm2 and cdk4 in one of six of these patients and showed positivity for CD34 in another one. Comparative genomic hybridization analysis performed on three samples from two of these patients showed no amplification of the 12q13-15 region but showed complex profiles. This study shows that most so-called malignant fibrous histiocytomas developed in the retroperitoneum are dedifferentiated liposarcoma and that a poorly differentiated sarcoma in this area should prompt extensive sampling to demonstrate a well-differentiated liposarcoma component, immunohistochemistry for mdm2 and cdk4, and if possible, a cytogenetic or a molecular biology analysis. PMID- 12640105 TI - KI-67 AND hTERT expression can aid in the distinction between malignant and benign pheochromocytoma and paraganglioma. AB - The clinical and histopathological distinction between benign and malignant pheochromocytomas and paragangliomas is difficult, and reliable diagnostic markers are lacking. Here we have evaluated the prognostic value of human telomerase reverse transcriptase (hTERT) gene expression detected by reverse transcription PCR (RT-PCR); telomerase activity (TA) measured by TRAP (telomeric repeat amplification protocol) assay; immunohistochemical staining for Ki-67/MIB 1; and the mRNA expression of matrix metalloproteinase (MMP)-2 and EMMPRIN (extracellular matrix metalloproteinase inducer) analyzed by in situ hybridization in 32 primary pheochromocytomas or abdominal paragangliomas. hTERT was expressed in 7/11 malignant tumors (defined as presence of metastasis and/or extensive local invasion) as compared with in 2/21 benign tumors. All of the benign tumors showed <1% proliferative activity, as measured by Ki-67/MIB-1 staining. In all three patients with malignant tumors who developed metastases and/or invasive local recurrence during follow-up, the tumors were positive for either hTERT expression or Ki-67/MIB-1 immunoreactivity. TA was not a significant discriminator between benign and malignant tumors, and the value of EMMPRIN and MMP-2 as predictive markers was limited. In conclusion, the findings imply that the combined use of Ki-67/MIB-1 and hTERT, in addition to histopathology, provides a highly specific tool to identify benign pheochromocytoma and abdominal paraganglioma cases that are not at risk of developing recurrent or metastatic disease. PMID- 12640107 TI - Oral and salivary gland angiosarcoma: a clinicopathologic study of 29 cases. AB - Angiosarcomas of the oral and salivary gland area are extremely rare, mostly presented as case reports. We wanted to study the clinicopathologic features of a series of oral and salivary gland angiosarcomas. Cases coded as "angiosarcoma" were retrieved from the Oral and Maxillofacial Pathology Department of the Armed Forces Institute of Pathology. Patient folders and pathology were reviewed and recorded; immunohistochemistry and follow-up were obtained. Inclusion required oral or salivary gland location, vasoformative growth, cytologic atypia, mitoses, and vascular markers. Skin, bone, and subcutaneous angiosarcomas were excluded. Primary and secondary (metastatic) oral angiosarcomas were included. The 22 primary angiosarcomas involved tongue (n = 9), parotid (n = 4), lip (n = 4), submandibular gland (n = 3), and 1 each of soft and hard palate. The 7 secondary angiosarcomas involved the gingiva (n = 4) and parotid gland (n = 3). Overall, patient ages ranged from 6-90 years (mean, 55 years). There were 15 males and 14 females. Symptoms included a mass with recent enlargement and bleeding. Tumor sizes ranged from 0.8-7.0 cm (mean, 2.6 cm). Histologically, all tumors were vasoformative; 86% had solid and 17% had distinctive papillary areas. Eight (28%) were classified as the epithelioid subtype. Immunohistochemical stains showed that the tumor cells were positive for Factor VIIIrag in 19/21, CD31 in 16/19, CD34 in 7/12, and Ulex in 1/1. Primary tumors were classified as low grade (n = 7, in all locations except salivary gland), intermediate (n = 7), and high grade (n = 8); all secondary tumors were high grade. Follow-up was available on 14/22 primary and 7/7 secondary angiosarcomas. Of primary tumors, two tongue angiosarcoma patients died at 1 and 9 years, but 4 were alive without disease over a mean of 7.3 years (range, 1-13 years). Four primary salivary gland angiosarcoma patients were alive without disease over a mean of 5.8 years (range, 1-14 years), and 1 had only a late (15 years) metastasis and death (at 20 years). Three primary lip angiosarcoma patients were without disease over a mean of 14.3 years (range, 13-16 years). Of secondary tumors, three salivary gland angiosarcoma patients died within 1 year, and all four secondary gingival angiosarcoma patients died of disease within 3 years. Assessing follow-up of primary oral and salivary gland angiosarcoma patients by grade, 5 patients with high-grade tumors had no evidence of disease over a mean of 7.6 years (range, 1 16 years), 3 patients with intermediate-grade tumors had no evidence of disease over a mean of 12.7 years (range, 11-14 years), 2 patients with intermediate grade tumors died of disease at 9 and 20 years, 3 patients with low-grade tumors had no evidence of disease over a mean of 6.3 years (range, 1-14 years), and 1 patient with low-grade tumor died of disease at 1 year. Primary oral and salivary gland angiosarcomas, albeit rare, mostly involve the tongue, parotid gland, and lip of adults, often with relatively good outcome. Although the most common angiosarcoma morphology in this area is spindled vasoformative and solid, almost one third of oral and salivary gland angiosarcomas are the rare epithelioid angiosarcoma variant. Most gingival and few parotid angiosarcomas appear to be metastases from other locations, with many patients succumbing to death within 3 years. Despite predominantly high- or intermediate-grade morphology, patients with primary angiosarcoma of the tongue, salivary gland, and lip have a better prognosis than do patients with primary cutaneous or deep soft tissue angiosarcoma, including those patients with secondary oral and salivary gland involvement. PMID- 12640108 TI - Role of the DPC4 tumor suppressor gene in adenocarcinoma of the ampulla of Vater: analysis of 140 cases. AB - The K-ras oncogene is activated in approximately 90% of pancreatic adenocarcinomas, and the DPC4 (MADH4/SMAD4) tumor suppressor gene is inactivated in approximately 55% of pancreatic adenocarcinomas. The contributions of these genetic alterations to the development of adenocarcinoma of the ampulla of Vater have not been fully established. One hundred forty surgically resected ampullary adenocarcinomas (76 with associated adenomas with high-grade dysplasia) were immunohistochemically labeled for the DPC4 gene product, and in 85 cases the results were correlated with the status of the K-ras oncogene from previously reported data. The results were correlated with clinical outcome and with other pathologic predictors of prognosis. Complete loss of Dpc4 labeling was identified in 34% (95% confidence interval [CI]: 26%, 43%) of the invasive carcinomas and in none (upper 95% CI: 6%) of the associated adenomas. Focal loss of Dpc4 was seen in three (4%; 95% CI: 1%, 14%) of the areas of high-grade dysplasia. Complete loss of Dpc4 expression was seen in 28/77 intestinal-type tumors, in 17/46 pancreaticobiliary-type tumors, and in 0/10 colloid carcinomas. Activating point mutations in the K-ras gene were identified in 40% of the invasive cancers. There was no correlation between K-ras gene mutations and Dpc4 expression and no correlation between these variables and survival. The overall 5-year survival rate was 38%. Lymph node metastases were associated with shorter survival (P =.03). Loss of Dpc4 expression occurs in approximately one third of invasive ampullary cancers but is not seen in adenomas; thus, loss of Dpc4 expression occurs late in ampullary carcinogenesis. Although ampullary and pancreatic adenocarcinomas share histologic and molecular features, ampullary carcinomas are less likely to show loss of Dpc4 expression or K-ras gene mutations. PMID- 12640109 TI - Negative regulation of BRCA1 gene expression by HMGA1 proteins accounts for the reduced BRCA1 protein levels in sporadic breast carcinoma. AB - A drastic reduction in BRCA1 gene expression is a characteristic feature of aggressive sporadic breast carcinoma. However, the mechanisms underlying BRCA1 downregulation in breast cancer are not well understood. Here we report that both in vitro and in vivo HMGA1b protein binds to and inhibits the activity of both human and mouse BRCA1 promoters. Consistently, murine embryonic stem (ES) cells with the Hmga1 gene deleted display higher Brca1 mRNA and protein levels than do wild-type ES cells. Stable transfection of MCF-7 cells with the HMGA1b cDNA results in a decrease of BRCA1 gene expression and in a lack of BRCA1 induction after estrogen treatment. Finally, we found an inverse correlation between HMGA1 and BRCA1 mRNA and protein expression in human mammary carcinoma cell lines and tissues. These data indicate that HMGA1 proteins are involved in transcriptional regulation of the BRCA1 gene, and their overexpression may have a role in BRCA1 downregulation observed in aggressive mammary carcinomas. PMID- 12640110 TI - Mitochondrial protein import: recognition of internal import signals of BCS1 by the TOM complex. AB - BCS1, a component of the inner membrane of mitochondria, belongs to the group of proteins with internal, noncleavable import signals. Import and intramitochondrial sorting of BCS1 are encoded in the N-terminal 126 amino acid residues. Three sequence elements were identified in this region, namely, the transmembrane domain (amino acid residues 51 to 68), a presequence type helix (residues 69 to 83), and an import auxiliary region (residues 84 to 126). The transmembrane domain is not required for stable binding to the TOM complex. The Tom receptors (Tom70, Tom22 and Tom20), as determined by peptide scan analysis, interact with the presequence-like helix, yet the highest binding was to the third sequence element. We propose that the initial recognition of BCS1 precursor at the surface of the organelle mainly depends on the auxiliary region and does not require the transmembrane domain. This essential region represents a novel type of signal with targeting and sorting functions. It is recognized by all three known mitochondrial import receptors, demonstrating their capacity to decode various targeting signals. We suggest that the BCS1 precursor crosses the TOM complex as a loop structure and that once the precursor emerges from the TOM complex, all three structural elements are essential for the intramitochondrial sorting to the inner membrane. PMID- 12640111 TI - Lysyl oxidase inhibits ras-mediated transformation by preventing activation of NF kappa B. AB - Lysyl oxidase (LO), which catalyzes the oxidation of lysine residues, was previously shown to have anti-oncogenic activity on ras-transformed cells. Since oncogenic Ras mediates transformation, in part, through the activation of the transcription factor nuclear factor-kappa B (NF-kappa B), we tested here the effects of LO on NF-kappa B activity. Expression of LO in ras-transformed NIH 3T3 cells led to decreased NF-kappa B binding and activity, as well as the expression of the NF-kappa B target gene c-myc. Importantly, ectopic expression of LO led to a dramatic decrease in colony formation by ras-transformed NIH 3T3 cells, a finding comparable to the expression of the I kappa B alpha dominant-negative mutant, which could be rescued by p65/p50 NF-kappa B subunit expression. LO was unable to directly inhibit the activity of ectopically expressed p65 and c-Rel NF kappa B subunits, suggesting that LO affected an upstream signaling pathway(s) induced by Ras. Consistent with this hypothesis, LO expression decreased both the rate of I kappa B alpha turnover and the activities of IKK alpha and IKK beta. Moreover, the ectopic expression of a constitutively active version of either kinase reversed the negative effects of LO. Ras can induce NF-kappa B via both the phosphatidylinositol 3-kinase (PI3K)/Akt and Raf/MEK pathways. LO potently downregulated the PI3K and Akt kinases, while partially inhibiting MEK kinase activity. Expression of a constitutively activated, myristylated Akt or PDK1 was able to counteract the effect of LO on NF-kappa B, whereas constitutively activated Raf was only partially effective. Importantly, LO blocked membrane localization of Akt and PDK1 in Ras-transformed cells. Overall, these results strongly argue that the anti-oncogenic effects of LO on ras-mediated transformation are due to its ability to inhibit signaling pathways that lead to activation of NF-kappa B. PMID- 12640112 TI - The Delta Np63 alpha phosphoprotein binds the p21 and 14-3-3 sigma promoters in vivo and has transcriptional repressor activity that is reduced by Hay-Wells syndrome-derived mutations. AB - p63 is a recently identified homolog of p53 that is found in the basal layer of several stratified epithelial tissues such as the epidermis, oral mucosa, prostate, and urogenital tract. Studies with p63(-/-) mice and analysis of several human autosomal-dominant disorders with germ line p63 mutations suggest p63 involvement in maintaining epidermal stem cell populations. The p63 gene encodes six splice variants with reported transactivating or dominant-negative activities. The goals of the current study were to determine the splice variants that are expressed in primary human epidermal keratinocytes (HEKs) and the biochemical activity p63 has in these epithelial cell populations. We found that the predominant splice variant expressed in HEKs was Delta Np63 alpha, and it was present as a phosphorylated protein. During HEK differentiation, Delta Np63 alpha and p53 levels decreased, while expression of p53 target genes p21 and 14-3-3 sigma increased. Delta Np63 alpha had transcriptional repressor activity in vitro, and this activity was reduced in Delta Np63 alpha proteins containing point mutations, corresponding to those found in patients with Hay-Wells syndrome. Further, we show that Delta Np63 alpha and p53 can bind the p21 and 14 3-3 sigma promoters in vitro and in vivo, with decreased binding of p63 to these promoters during HEK differentiation. These data suggest that Delta Np63 alpha acts as a transcriptional repressor at select growth regulatory gene promoters in HEKs, and this repression likely plays an important role in the proliferative capacity of basal keratinocytes. PMID- 12640114 TI - p120 Catenin-associated Fer and Fyn tyrosine kinases regulate beta-catenin Tyr 142 phosphorylation and beta-catenin-alpha-catenin Interaction. AB - beta-Catenin has a key role in the formation of adherens junction through its interactions with E-cadherin and alpha-catenin. We show here that interaction of beta-catenin with alpha-catenin is regulated by the phosphorylation of beta catenin Tyr-142. This residue can be phosphorylated in vitro by Fer or Fyn tyrosine kinases. Transfection of these kinases to epithelial cells disrupted the association between both catenins. We have also examined whether these kinases are involved in the regulation of this interaction by K-ras. Stable transfectants of the K-ras oncogene in intestinal epithelial IEC18 cells were generated which show little alpha-catenin-beta-catenin association with respect to control clones; this effect is accompanied by increased Tyr-142 phosphorylation and activation of Fer and Fyn kinases. As reported for Fer, Fyn kinase is constitutively bound to p120 catenin; expression of K-ras induces the phosphorylation of p120 catenin on tyrosine residues increasing its affinity for E-cadherin and, consequently, promotes the association of Fyn with the adherens junction complex. Yes tyrosine kinase also binds to p120 catenin but only upon activation, and stimulates Fer and Fyn tyrosine kinases. These results indicate that p120 catenin acts as a docking protein facilitating the activation of Fer/Fyn tyrosine kinases by Yes and demonstrate the role of these p120 catenin associated kinases in the regulation of beta-catenin-alpha-catenin interaction. PMID- 12640113 TI - Retinoic acid repression of bone morphogenetic protein 4 in inner ear development. AB - Bone morphogenetic protein 4 (BMP4) and retinoic acid are important for normal development of the inner ear, but whether they are linked mechanistically is not known. BMP4 antagonists disrupt semicircular canal formation, as does exposure to retinoic acid. We demonstrate that retinoic acid directly down-regulates BMP4 transcription in a mouse inner ear-derived cell line, and we identify a novel promoter in the second intron of the BMP4 gene that is a target of this regulation both in the cell line and in the mouse embryonic inner ear in vivo. The importance of this down-regulation is demonstrated in chicken embryos by showing that the retinoic acid effect on semicircular canal development can be overcome by exogenous BMP4. PMID- 12640116 TI - Polymerase mu is a DNA-directed DNA/RNA polymerase. AB - DNA polymerases are defined as such because they use deoxynucleotides instead of ribonucleotides with high specificity. We show here that polymerase mu (pol mu), implicated in the nonhomologous end-joining pathway for repair of DNA double strand breaks, incorporates both ribonucleotides and deoxynucleotides in a template-directed manner. pol mu has an approximately 1,000-fold-reduced ability to discriminate against ribonucleotides compared to that of the related pol beta, although pol mu's substrate specificity is similar to that of pol beta in most other respects. Moreover, pol mu more frequently incorporates ribonucleotides when presented with nucleotide concentrations that approximate cellular pools. We therefore addressed the impact of ribonucleotide incorporation on the activities of factors required for double-strand break repair by nonhomologous end joining. We determined that the ligase required for this pathway readily joined strand breaks with terminal ribonucleotides. Most significantly, pol mu frequently introduced ribonucleotides into the repair junctions of an in vitro nonhomologous end-joining reaction, an activity that would be expected to have important consequences in the context of cellular double-strand break repair. PMID- 12640115 TI - MEK kinase 2 and the adaptor protein Lad regulate extracellular signal-regulated kinase 5 activation by epidermal growth factor via Src. AB - Lad is an SH2 domain-containing adaptor protein that binds MEK kinase 2 (MEKK2), a mitogen-activated protein kinase (MAPK) kinase kinase for the extracellular signal-regulated kinase 5 (ERK5) and JNK pathways. Lad and MEKK2 are in a complex in resting cells. Antisense knockdown of Lad expression and targeted gene disruption of MEKK2 expression results in loss of epidermal growth factor (EGF) and stress stimuli-induced activation of ERK5. Activation of MEKK2 and the ERK5 pathway by EGF and stress stimuli is dependent on Src kinase activity. The Lad binding motif is encoded within amino acids 228 to 282 in the N terminus of MEKK2, and expression of this motif blocks Lad-MEKK2 interaction, resulting in inhibition of Src-dependent activation of MEKK2 and ERK5. JNK activation by EGF is similarly inhibited by loss of Lad or MEKK2 expression and by blocking the interaction of MEKK2 and Lad. Our studies demonstrate that Src kinase activity is required for ERK5 activation in response to EGF, MEKK2 expression is required for ERK5 activation by Src, Lad and MEKK2 association is required for Src activation of ERK5, and EGF and Src stimulation of ERK5-regulated MEF2-dependent promoter activity requires a functional Lad-MEKK2 signaling complex. PMID- 12640117 TI - von Hippel-Lindau protein-mediated repression of tumor necrosis factor alpha translation revealed through use of cDNA arrays. AB - Based on evidence that the von Hippel-Lindau (VHL) tumor suppressor protein is associated with polysomes and interacts with translation regulatory factors, we set out to investigate the potential influence of pVHL on protein translation. To this end, renal cell carcinoma (RCC) cells that either lacked pVHL or expressed pVHL through stable transfection were used to prepare RNA from cytosolic (unbound) and polysome-bound fractions. Hybridization of cDNA arrays using RNA from each fraction revealed a subset of transcripts whose abundance in polysomes decreased when pVHL function was restored. The tumor necrosis factor alpha (TNF alpha) mRNA was identified as one of the transcripts that preferentially associated with polysomes in pVHL-deficient cells. Additional evidence that the TNF-alpha mRNA was a target of translational repression by pVHL was obtained from reporter gene assays, which further revealed that pVHL's inhibitory influence on protein synthesis occurred through the TNF-alpha 3'-untranslated region. Our findings uncover a novel function for the pVHL tumor suppressor protein as regulator of protein translation. PMID- 12640118 TI - HMG-I(Y) and the CBP/p300 coactivator are essential for human papillomavirus type 18 enhanceosome transcriptional activity. AB - A strong epithelial specific enhancer drives transcription of the human papillomavirus type 18 (HPV18) oncogenes. Its activity depends on the formation of a higher-order nucleoprotein complex (enhanceosome) involving the sequence specific JunB/Fra2 transcription factor and the HMG-I(Y) architectural protein. Here we show that proteins from HeLa cell nuclear extract cover almost all of the HPV18 enhancer sequences and that it contains seven binding sites for the purified HMG-I(Y) protein, providing evidence for a tight nucleoprotein structure. Binding of HMG-I(Y) and the AP1 heterodimer from HeLa nuclear extract to overlapping sites of the core enhanceosome is cooperative. The integrity of this specific HMG-I(Y) binding site is as essential as the AP1 binding site for the enhancer function, indicating the fundamental role played by this architectural protein. We demonstrate that the CBP/p300 coactivator is recruited by the HPV18 enhanceosome and that it is limiting for transcriptional activation, since it is sequestered by the adenovirus E1A protein and by the JunB/Fra2 positive factor in excess. We show the involvement of JunB and p300 in vivo in the HPV18 transcription by chromatin immunoprecipitation of HPV18 sequences in HeLa cells. PMID- 12640119 TI - Transcription-dependent degradation of topoisomerase I-DNA covalent complexes. AB - Topoisomerase I (Top I)-DNA covalent complexes represent a unique type of DNA lesion whose repair and processing remain unclear. In this study, we show that Top I-DNA covalent complexes transiently arrest RNA transcription in normal nontransformed cells. Arrest of RNA transcription is coupled to activation of proteasomal degradation of Top I and the large subunit of RNA polymerase II. Recovery of transcription occurs gradually and depends on both proteasomal degradation of Top I and functional transcription-coupled repair (TCR). These results suggest that arrest of the RNA polymerase elongation complex by the Top I DNA covalent complex triggers a 26S proteasome-mediated signaling pathway(s) leading to degradation of both Top I and the large subunit of RNA polymerase II. We propose that proteasomal degradation of Top I and RNA polymerase II precedes repair of the exposed single-strand breaks by TCR. PMID- 12640120 TI - Commitment point during G0-->G1 that controls entry into the cell cycle. AB - Initiation of T-lymphocyte-mediated immune responses involves two cellular processes: entry into the cell cycle (G(0)-->G(1)) for clonal proliferation and coordinated changes in surface and secreted molecules that mediate effector functions. However, a point during G(0)-->G(1) beyond which T cells are committed to enter the cell cycle has not been defined. We define here a G(0)-->G(1) commitment point that occurs 3 to 5 h after CD3 and CD28 stimulation of human CD4 or CD8 T cells. Transition through this point requires cdk6/4-cyclin D, since inhibition with TAT-p16(INK4A) during the first 3 to 5 h prevents cell cycle entry and maintains both naive and memory T cells in G(0). Transition through the G(0)-->G(1) commitment point is also necessary for T cells to increase in size, i.e., to enter the cellular growth cycle. However, transition through this point is not required for the induction of effector functions. These can be initiated while cells are maintained in G(0) with TAT-p16(INK4A). We have termed this quiescent, activated state G(0(A)). Our data provide proof of the principle that entry of T cells into the cell cycle and cellular growth cycles are coupled at the G(0)-->G(1) commitment point but that these processes can be uncoupled from the early expression of molecules of effector functions. PMID- 12640122 TI - Synthesis of signals for de novo DNA methylation in Neurospora crassa. AB - Most 5-methylcytosine in Neurospora crassa occurs in A:T-rich sequences high in TpA dinucleotides, hallmarks of repeat-induced point mutation. To investigate how such sequences induce methylation, we developed a sensitive in vivo system. Tests of various 25- to 100-bp synthetic DNA sequences revealed that both T and A residues were required on a given strand to induce appreciable methylation. Segments composed of (TAAA)(n) or (TTAA)(n) were the most potent signals; 25-mers induced robust methylation at the special test site, and a 75-mer induced methylation elsewhere. G:C base pairs inhibited methylation, and cytosines 5' of ApT dinucleotides were particularly inhibitory. Weak signals could be strengthened by extending their lengths. A:T tracts as short as two were found to cooperate to induce methylation. Distamycin, which, like the AT-hook DNA binding motif found in proteins such as mammalian HMG-I, binds to the minor groove of A:T rich sequences, suppressed DNA methylation and gene silencing. We also found a correlation between the strength of methylation signals and their binding to an AT-hook protein (HMG-I) and to activities in a Neurospora extract. We propose that de novo DNA methylation in Neurospora cells is triggered by cooperative recognition of the minor groove of multiple short A:T tracts. Similarities between sequences subjected to repeat-induced point mutation in Neurospora crassa and A:T-rich repeated sequences in heterochromatin in other organisms suggest that related mechanisms control silent chromatin in fungi, plants, and animals. PMID- 12640121 TI - Manganese superoxide dismutase-mediated gene expression in radiation-induced adaptive responses. AB - Antioxidant enzymes are critical in oxidative stress responses. Radioresistant variants isolated from MCF-7 human carcinoma cells following fractionated ionizing radiation (MCF+FIR cells) or overexpression of manganese superoxide dismutase (MCF+SOD cells) demonstrated dose-modifying factors at 10% isosurvival of 1.8 and 2.3, respectively. MCF+FIR and MCF-7 cells (exposed to single-dose radiation) demonstrated 5- to 10-fold increases in MnSOD activity, mRNA, and immunoreactive protein. Radioresistance in MCF+FIR and MCF+SOD cells was reduced following expression of antisense MnSOD. DNA microarray analysis and immunoblotting identified p21, Myc, 14-3-3 zeta, cyclin A, cyclin B1, and GADD153 as genes constitutively overexpressed (2- to 10-fold) in both MCF+FIR and MCF+SOD cells. Radiation-induced expression of these six genes was suppressed in fibroblasts from Sod2 knockout mice (-/-) as well as in MCF+FIR and MCF+SOD cells expressing antisense MnSOD. Inhibiting NF-kappa B transcriptional activity in MCF+FIR cells, by using mutant I kappa B alpha, inhibited radioresistance as well as reducing steady-state levels of MnSOD, 14-3-3 zeta, GADD153, cyclin A, and cyclin B1 mRNA. In contrast, mutant I kappa B alpha was unable to inhibit radioresistance or reduce 14-3-3 zeta, GADD153, cyclin A, and cyclin B1 mRNAs in MCF+SOD cells, where MnSOD overexpression was independent of NF-kappa B. These results support the hypothesis that NF-kappa B is capable of regulating the expression of MnSOD, which in turn is capable of increasing the expression of genes that participate in radiation-induced adaptive responses. PMID- 12640124 TI - Caveolin-induced activation of the phosphatidylinositol 3-kinase/Akt pathway increases arsenite cytotoxicity. AB - The inhibitory effect of caveolin on the cellular response to growth factor stimulation is well established. Given the significant overlap in signaling pathways involved in regulating cell proliferation and stress responsiveness, we hypothesized that caveolin would also affect a cell's ability to respond to environmental stress. Here we investigated the ability of caveolin-1 to modulate the cellular response to sodium arsenite and thereby alter survival of the human cell lines 293 and HeLa. Cells stably transfected with caveolin-1 were found to be much more sensitive to the toxic effects of sodium arsenite than either untransfected parental cells or parental cells transfected with an empty vector. Unexpectedly, the caveolin-overexpressing cells also exhibited a significant activation of the phosphatidylinositol 3-kinase (PI3K)/Akt pathway, which additional studies suggested was likely due to decreased neutral sphingomyelinase activity and ceramide synthesis. In contrast to its extensively documented antiapoptotic influence, the elevated activity of Akt appears to be important in sensitizing caveolin-expressing cells to arsenite-induced toxicity, as both pretreatment of cells with the PI3K inhibitor wortmannin and overexpression of a dominant-negative Akt mutant markedly improved the survival of arsenite-treated cells. This death-promoting influence of the PI3K/Akt pathway in caveolin overexpressing cells appeared not to be unique to sodium arsenite, as wortmannin pretreatment also resulted in increased survival in the presence of H(2)O(2). In summary, our results indicate that caveolin-induced upregulation of the PI3K/Akt signaling pathway, which appears to be a death signal in the presence of arsenite and H(2)O(2), sensitizes cells to environmental stress. PMID- 12640123 TI - Structural requirements of SLP-76 in signaling via the high-affinity immunoglobulin E receptor (Fc epsilon RI) in mast cells. AB - The adapter SLP-76 plays an essential role in Fc epsilon RI signaling, since SLP 76(-/-) bone marrow-derived mast cells (BMMC) fail to degranulate and release interleukin-6 (IL-6) following Fc epsilon RI ligation. To define the role of SLP 76 domains and motifs in Fc epsilon RI signaling, SLP-76(-/-) BMMC were retrovirally transduced with SLP-76 and SLP-76 mutants. The SLP-76 N-terminal and Gads binding domains, but not the SH2 domain, were critical for Fc epsilon RI mediated degranulation and IL-6 secretion, whereas all three domains are essential for T-cell proliferation following T-cell receptor (TCR) ligation. Unexpectedly, the three tyrosine residues in SLP-76 critical for TCR signaling, Y112, Y128, and Y145, were not essential for IL-6 secretion, but were required for degranulation and mitogen-activated protein kinase activation. Furthermore, a Y112/128F SLP-76 mutant, but not a Y145F mutant, strongly reconstituted mast cell degranulation, suggesting a critical role for Y145 in Fc epsilon RI-mediated exocytosis. These results point to important differences in the function of SLP 76 between T cells and mast cells. PMID- 12640125 TI - Gab3-deficient mice exhibit normal development and hematopoiesis and are immunocompetent. AB - Gab proteins are intracellular scaffolding and docking molecules involved in signaling pathways mediated by various growth factor, cytokine, or antigen receptors. Gab3 has been shown to act downstream of the macrophage colony stimulating factor receptor, c-Fms, and to be important for macrophage differentiation. To analyze the physiological role of Gab3, we used homologous recombination to generate mice deficient in Gab3. Gab3(-/-) mice develop normally, are visually indistinguishable from their wild-type littermates, and are healthy and fertile. To obtain a detailed expression pattern of Gab3, we generated Gab3-specific monoclonal antibodies. Immunoblotting revealed a predominant expression of Gab3 in lymphocytes and bone marrow-derived macrophages. However, detailed analysis demonstrated that hematopoiesis in mice lacking Gab3 is not impaired and that macrophages develop in normal numbers and exhibit normal function. The lack of Gab3 expression during macrophage differentiation is not compensated for by increased levels of Gab1 or Gab2 mRNA. Furthermore, Gab3-deficient mice have no major immune deficiency in T- and B lymphocyte responses to protein antigens or during viral infection. In addition, allergic responses in Gab3-deficient mice appeared to be normal. Together, these data demonstrate that loss of Gab3 does not result in detectable defects in normal mouse development, hematopoiesis, or immune system function. PMID- 12640126 TI - Myocardin is a critical serum response factor cofactor in the transcriptional program regulating smooth muscle cell differentiation. AB - The SAP family transcription factor myocardin functionally synergizes with serum response factor (SRF) and plays an important role in cardiac development. To determine the function of myocardin in the smooth muscle cell (SMC) lineage, we mapped the pattern of myocardin gene expression and examined the molecular mechanisms underlying transcriptional activity of myocardin in SMCs and embryonic stem (ES) cells. The human and murine myocardin genes were expressed in vascular and visceral SMCs at levels equivalent to or exceeding those observed in the heart. During embryonic development, the myocardin gene was expressed abundantly in a precise, developmentally regulated pattern in SMCs. Forced expression of myocardin transactivated multiple SMC-specific transcriptional regulatory elements in non-SMCs. By contrast, myocardin-induced transactivation was not observed in SRF(-/-) ES cells but could be rescued by forced expression of SRF or the SRF DNA-binding domain. Furthermore, expression of a dominant-negative myocardin mutant protein or small-interfering-RNA-induced myocardin knockdown significantly reduced SM22 alpha promoter activity in SMCs. Most importantly, forced expression of myocardin activated expression of the SM22 alpha, smooth muscle alpha-actin, and calponin-h1 genes in undifferentiated mouse ES cells. Taken together, these data demonstrate that myocardin plays an important role in the SRF-dependent transcriptional program that regulates SMC development and differentiation. PMID- 12640127 TI - Changes in histone acetylation are associated with differences in accessibility of V(H) gene segments to V-DJ recombination during B-cell ontogeny and development. AB - Although V(D)J recombination is thought to be regulated by changes in the accessibility of chromatin to the recombinase machinery, the mechanisms responsible for establishing "open" chromatin are poorly understood. We performed a detailed study of the acetylation status of histones associated with 11 V(H) gene segments, their flanking regions, and various intergenic elements during B cell development and ontogeny, when V(D)J recombination is highly regulated. Histone H4 shows higher and more-regulated acetylation than does histone H3 in the V(H) locus. In adult pro-B cells, V(H) gene segments are acetylated prior to V(D)J rearrangement, with higher acetylation associated with J(H)-distal V(H) gene segments. While large regions of the V(H) locus have similar patterns of histone acetylation, acetylation is narrowly confined to the gene segments, their flanking promoters, and recombinase signal sequence elements. Thus, histone acetylation in the V(H) locus is both locally and globally regulated. Increased histone acetylation accompanies preferential recombination of J(H)-proximal V(H) gene segments in early B-cell ontogeny, and decreased histone acetylation accompanies inhibition of V-DJ recombination in a transgenic model of immunoglobulin heavy-chain allelic exclusion. Thus, changes in histone acetylation appear to be important for both promotion and inhibition of V-DJ rearrangement during B-cell ontogeny and development. PMID- 12640128 TI - Involvement of poly(ADP-Ribose) polymerase 1 and poly(ADP-Ribosyl)ation in regulation of centrosome function. AB - The regulatory mechanism of centrosome function is crucial to the accurate transmission of chromosomes to the daughter cells in mitosis. Recent findings on the posttranslational modifications of many centrosomal proteins led us to speculate that these modifications might be involved in centrosome behavior. Poly(ADP-ribose) polymerase 1 (PARP-1) catalyzes poly(ADP-ribosyl)ation to various proteins. We show here that PARP-1 localizes to centrosomes and catalyzes poly(ADP-ribosyl)ation of centrosomal proteins. Moreover, centrosome hyperamplification is frequently observed with PARP inhibitor, as well as in PARP 1-null cells. Thus, it is possible that chromosomal instability known in PARP-1 null cells can be attributed to the centrosomal dysfunction. P53 tumor suppressor protein has been also shown to be localized at centrosomes and to be involved in the regulation of centrosome duplication and monitoring of the chromosomal stability. We found that centrosomal p53 is poly(ADP-ribosyl)ated in vivo and centrosomal PARP-1 directly catalyzes poly(ADP-ribosyl)ation of p53 in vitro. These results indicate that PARP-1 and PARP-1-mediated poly(ADP-ribosyl)ation of centrosomal proteins are involved in the regulation of centrosome function. PMID- 12640129 TI - Supramolecular complex formation between Rad6 and proteins of the p53 pathway during DNA damage-induced response. AB - The HR6A and -B genes, homologues of the yeast Rad6 gene, encode ubiquitin conjugating enzymes that are required for postreplication repair of DNA and damage-induced mutagenesis. Using surface plasmon resonance, we show here that HR6 protein (referred as Rad6) physically interacts with p53. Analysis of proteins coimmunoprecipitated with Rad6 antibody from metabolically labeled normal MCF10A human breast epithelial cells not only confirmed Rad6-p53 interactions in vivo but also demonstrated for the first time that exposure of MCF10A cells to cisplatin or adriamycin (ADR) induces recruitment of p14ARF into Rad6-p53 complexes. Further analysis of ADR-induced p53 response showed that stable Rad6-p53-p14ARF complex formation is associated with a parallel increase and decrease in monoubiquitinated and polyubiquitinated p53, respectively, and arrest in G(2)/M phase of the cell cycle. Interestingly, the ADR-induced suppression of p53 polyubiquitination correlated with a corresponding decline in intact Hdm2 protein levels. Treatment of MCF10A cells with MG132, a 26S proteasome inhibitor, effectively stabilized monoubiquitinated p53 and rescued ADR-induced downregulation of Hdm2. These data suggest that ADR-induced degradation of Hdm2 occurs via the ubiquitin-proteasome pathway. Rad6 is present in both the cytoplasmic and nuclear compartments of normal MCF10A cells, although in response to DNA damage it is predominantly found in the nucleus colocalizing with ubiquitinated p53, whereas Hdm2 is undetectable. Consistent with in vivo data, results from in vitro ubiquitination assays show that Rad6 mediates addition of one (mono-) to two (multimono-) ubiquitin molecules on p53 and that inclusion of Mdm2 is essential for its polyubiquitination. The data presented in the present study suggest that Rad6-p53-p14ARF complex formation and p53 ubiquitin modification are important damage-induced responses that perhaps determine the fidelity of DNA postreplication repair. PMID- 12640131 TI - Functional analysis of basic transcription element binding protein by gene targeting technology. AB - Basic transcription element binding protein (BTEB) is a transcription factor with a characteristic zinc finger motif and is most remarkably enhanced by thyroid hormone T(3) treatment (R. J. Denver et al., J. Biol. Chem. 272:8179-8188, 1997). To investigate the function of BTEB per se and to touch on the effects of T(3) (3,5,3'-triiodothyronine) on mouse development, we generated BTEB-deficient mice by gene knockout technology. Homologous BTEB(-/-) mutant mice were bred according to apparently normal Mendelian genetics, matured normally, and were fertile. Mutant mice could survive for at least 2 years without evident pathological defects. From the expression of lacZ, which was inserted into the reading frame of the BTEB gene, BTEB showed a characteristic tissue-specific expression profile during the developmental process of brain and bone. Dramatically increased expression of BTEB was observed in Purkinje cells of the cerebellum and pyramidal cell layers of the hippocampus at P7 when synapses start to form in the brain. Although general behavioral activities such as locomotion, rearing, and speed of movement were not so much affected in the BTEB(-/-) mutant mice, they showed clearly reduced activity levels in rotorod and contextual fear-conditioning tests; this finding was probably due to defective functions of the cerebellum, hippocampus, and amygdala. PMID- 12640133 TI - Gads/Grb2-mediated association with LAT is critical for the inhibitory function of Gab2 in T cells. AB - A docking protein, Gab2, is recruited to the vicinity of the TCR complex and inhibits downstream signaling by interaction with negative regulators. However, the molecular mechanisms of this recruitment remain unclear. We have found that Gab2 associates with LAT upon TCR stimulation and that LAT is essential for Gab2 phosphorylation. By analysis of several Gab2 mutants, the c-Met binding domain (MBD) of Gab2 was found to be both necessary and sufficient for stimulation induced LAT binding. Within the MBD domain, a novel Grb2 SH3 binding motif, PXXXR, is critical for constitutive association with Gads/Grb2. Through this association, Gab2 is recruited to the lipid raft after TCR ligation and exerts inhibitory function. The in vivo significance of this association is illustrated by the fact that T-cell responses are impaired in transgenic mice expressing wild type Gab2 but not in mice expressing mutant Gab2 lacking the motif. Furthermore, T cells from Gab2-deficient mice showed enhanced proliferative responses upon TCR stimulation. These results indicate that Gads/Grb2-mediated LAT association is critical for the inhibitory function of Gab2, implying that Gab2 induced in stimulated T cells may exert an efficient negative feedback loop by recruiting inhibitory molecules to the lipid raft and competing with SLP-76 through Gads binding. PMID- 12640130 TI - GGAPs, a new family of bifunctional GTP-binding and GTPase-activating proteins. AB - G proteins are molecular switches that control a wide variety of physiological functions, including neurotransmission, transcriptional activation, cell migration, cell growth. and proliferation. The ability of GTPases to participate in signaling events is determined by the ratio of GTP-bound to GDP-bound forms in the cell. All known GTPases exist in an inactive (GDP-bound) and an active (GTP bound) conformation, which are catalyzed by guanine nucleotide exchange factors and GTPase-activating proteins (GAPs), respectively. In this study, we identified and characterized a new family of bifunctional GTP-binding and GTPase-activating proteins, named GGAP. GGAPs contain an N-terminal Ras homology domain, called the G domain, followed by a pleckstrin homology (PH) domain, a C-terminal GAP domain, and a tandem ankyrin (ANK) repeat domain. Expression analysis indicates that this new family of proteins has distinct cell localization, tissue distribution, and even message sizes. GTPase assays demonstrate that GGAPs have high GTPase activity through direct intramolecular interaction of the N-terminal G domain and the C-terminal GAP domain. In the absence of the GAP domain, the N-terminal G domain has very low activity, suggesting a new model of GGAP protein regulation via intramolecular interaction like the multidomain protein kinases. Overexpression of GGAPs leads to changes in cell morphology and activation of gene transcription. PMID- 12640132 TI - Modulation of Rab5 and Rab7 recruitment to phagosomes by phosphatidylinositol 3 kinase. AB - Phagosomal biogenesis is central for microbial killing and antigen presentation by leukocytes. However, the molecular mechanisms governing phagosome maturation are poorly understood. We analyzed the role and site of action of phosphatidylinositol 3-kinases (PI3K) and of Rab GTPases in maturation using both professional and engineered phagocytes. Rab5, which is recruited rapidly and transiently to the phagosome, was found to be essential for the recruitment of Rab7 and for progression to phagolysosomes. Similarly, functional PI3K is required for successful maturation. Remarkably, inhibition of PI3K did not preclude Rab5 recruitment to phagosomes but instead enhanced and prolonged it. Moreover, in the presence of PI3K inhibitors Rab5 was found to be active, as deduced from measurements of early endosome antigen 1 binding and by photobleaching recovery determinations. Though their ability to fuse with late endosomes and lysosomes was virtually eliminated by wortmannin, phagosomes nevertheless recruited a sizable amount of Rab7. Moreover, Rab7 recruited to phagosomes in the presence of PI3K antagonists retained the ability to bind its effector, Rab7-interacting lysosomal protein, suggesting that it is functionally active. These findings imply that (i) dissociation of Rab5 from phagosomes requires products of PI3K, (ii) PI3K-dependent effectors of Rab5 are not essential for the recruitment of Rab7 by phagosomes, and (iii) recruitment and activation of Rab7 are insufficient to induce fusion of phagosomes with late endosomes and lysosomes. Accordingly, transfection of constitutively active Rab7 did not bypass the block of phagolysosome formation exerted by wortmannin. We propose that Rab5 activates both PI3K-dependent and PI3K-independent effectors that act in parallel to promote phagosome maturation. PMID- 12640134 TI - Dissecting the contribution of p16(INK4A) and the Rb family to the Ras transformed phenotype. AB - Although oncogenic Ras commonly contributes to the development of cancer, in normal primary cells it induces cell cycle arrest rather than transformation. Here we analyze the additional genetic changes required for Ras to promote cell cycle progression rather than arrest. We show that loss of p53 is sufficient for oncogenic Ras to stimulate proliferation in the absence of extrinsic mitogens in attached cells. However, surprisingly, we find that p53 loss is not sufficient for Ras to overcome anchorage dependence or contact inhibition. In contrast, expression of simian virus 40 (SV40) large T antigen (LT) allows Ras to overcome these additional cell cycle controls. Mutational analysis of SV40 LT shows that this action of SV40 LT depends on its ability to inactivate the retinoblastoma (Rb) family of proteins, in concert with the loss of p53. Importantly, we show that inactivation of the Rb family of proteins can be mimicked by loss of the cyclin-dependent kinase inhibitor p16(INK4A). p16(INK4A) is commonly lost in human tumors, but its contribution to the transformed phenotype is unknown. We demonstrate here a role for p16(INK4A) in the loss of cell cycle controls required for tumorigenesis and show how accumulating genetic changes cooperate and contribute to the transformed phenotype. PMID- 12640135 TI - Common interaction surfaces of the toll-like receptor 4 cytoplasmic domain stimulate multiple nuclear targets. AB - Toll-like receptor 4 (TLR4) mediates the host response to lipopolysaccharide (LPS) by promoting the activation of pro- and anti-inflammatory cytokine genes. To activate each gene, numerous signal transduction pathways are required. The adaptor proteins MyD88 and TIRAP contribute to the activation of several and possibly all pathways via direct interactions with TLR4's Toll/interleukin-1 receptor (IL-1R) (TIR) domain. However, additional adaptors that are required for the activation of specific subsets of pathways may exist, which could contribute to the differential regulation of target genes. Furthermore, it remains unknown whether direct interactions that have been reported between TIR domains and other proteins are required for TLR4 signaling. To address these issues, we systematically mutated the TLR4 TIR domain in the context of a CD4/TLR4 fusion protein. Several exposed residues defining at least two structural surfaces were required in macrophages for activation of the proinflammatory IL-12 p40 and anti inflammatory IL-10 promoters, as well as promoters dependent on individual transcription factors. Interestingly, the same residues were required by all promoters tested, suggesting that the signaling pathways diverge downstream of the adaptors. The mutant phenotypes provide a framework for future studies of TLR4 signaling, as the interaction supported by each critical surface residue will need to be defined. PMID- 12640136 TI - p53 Binding protein 53BP1 is required for DNA damage responses and tumor suppression in mice. AB - 53BP1 is a p53 binding protein of unknown function that binds to the central DNA binding domain of p53. It relocates to the sites of DNA strand breaks in response to DNA damage and is a putative substrate of the ataxia telangiectasia-mutated (ATM) kinase. To study the biological role of 53BP1, we disrupted the 53BP1 gene in the mouse. We show that, similar to ATM(-/-) mice, 53BP1-deficient mice were growth retarded, immune deficient, radiation sensitive, and cancer prone. 53BP1( /-) cells show a slight S-phase checkpoint defect and prolonged G(2)/M arrest after treatment with ionizing radiation. Moreover, 53BP1(-/-) cells feature a defective DNA damage response with impaired Chk2 activation. These data indicate that 53BP1 acts downstream of ATM and upstream of Chk2 in the DNA damage response pathway and is involved in tumor suppression. PMID- 12640137 TI - Myeloid cell function in MRP-14 (S100A9) null mice. AB - Myeloid-related protein 14 (MRP-14) and its heterodimeric partner, MRP-8, are cytosolic calcium-binding proteins, highly expressed in neutrophils and monocytes. To understand the function of MRP-14, we performed targeted disruption of the MRP-14 gene in mice. MRP-14(-/-) mice showed no obvious phenotype and were fertile. MRP-8 mRNA but not protein is present in the myeloid cells of these mice, suggesting that the stability of MRP-8 protein is dependent on MRP-14 expression. A compensatory increase in other proteins was not detected in cells lacking MRP-8 and MRP-14. Although the morphology of MRP-14(-/-) myeloid cells was not altered, they were significantly less dense. When Ca(2+) responses were investigated, there was no change in the maximal response to the chemokine MIP-2. At lower concentrations, however, there was reduced responsiveness in MRP-14(-/-) compared with MRP-14(+/+) neutrophils. This alteration in the ability to flux Ca(2+) did not impair the ability of the MRP-14(-/-) neutrophils to respond chemotactically to MIP-2. In addition, the myeloid cell functions of phagocytosis, superoxide burst, and apoptosis were unaffected in MRP-14(-/-) cells. In an in vivo model of peritonitis, MRP-14(-/-) mice showed no difference from wild-type mice in induced inflammatory response. The data indicate that MRP 14 and MRP-8 are dispensable for many myeloid cell functions. PMID- 12640139 TI - Coactivator-dependent acetylation stabilizes members of the SREBP family of transcription factors. AB - Members of the SREBP family of transcription factors control cholesterol and lipid homeostasis and play important roles during adipocyte differentiation. The transcriptional activity of SREBPs is dependent on the coactivators p300 and CBP. We now present evidence that SREBPs are acetylated by the intrinsic acetyltransferase activity of p300 and CBP. In SREBP1a, the acetylated lysine residue resides in the DNA-binding domain of the protein. Coexpression with p300 dramatically increases the expression of both SREBP1a and SREBP2, and this effect is dependent on the acetyltransferase activity of p300, indicating that acetylation of SREBPs regulates their stability. Indeed, acetylation or mutation of the acetylated lysine residue in SREBP1a stabilizes the protein. We demonstrate that the acetylated residue in SREBP1a is also targeted by ubiquitination and that acetylation inhibits this process. Thus, our studies define acetylation-dependent stabilization of transcription factors as a novel mechanism for coactivators to regulate gene expression. PMID- 12640138 TI - DNA hypermethylation in Drosophila melanogaster causes irregular chromosome condensation and dysregulation of epigenetic histone modifications. AB - The level of genomic DNA methylation plays an important role in development and disease. In order to establish an experimental system for the functional analysis of genome-wide hypermethylation, we overexpressed the mouse de novo methyltransferase Dnmt3a in Drosophila melanogaster. These flies showed severe developmental defects that could be linked to reduced rates of cell cycle progression and irregular chromosome condensation. In addition, hypermethylated chromosomes revealed elevated rates of histone H3-K9 methylation and a more restricted pattern of H3-S10 phosphorylation. The developmental and chromosomal defects induced by DNA hypermethylation could be rescued by mutant alleles of the histone H3-K9 methyltransferase gene Su(var)3-9. This mutation also resulted in a significantly decreased level of genomic DNA methylation. Our results thus uncover the molecular consequences of genomic hypermethylation and demonstrate a mutual interaction between DNA methylation and histone methylation. PMID- 12640141 TI - Mice lacking the 68-amino-acid, mammal-specific N-terminal extension of WT1 develop normally and are fertile. AB - Mutations in the Wilms' tumor 1 gene, WT1, cause pediatric nephroblastoma and the severe genitourinary disorders of Frasier and Denys-Drash syndromes. High levels of WT1 expression are found in the developing kidney, uterus, and testis- consistent with this finding, the WT1 knockout mouse demonstrates that WT1 is essential for normal genitourinary development. The WT1 gene encodes multiple isoforms of a zinc finger-containing protein by a combination of alternative splicing and alternative translation initiation. The use of an upstream, alternative CUG translation initiation codon specific to mammals results in the production of WT1 protein isoforms with a 68-amino-acid N-terminal extension. To determine the function in vivo of mammal-specific WT1 isoforms containing this extension, gene targeting was employed to introduce a subtle mutation into the WT1 gene. Homozygous mutant mice show a specific absence of the CUG-initiated WT1 isoforms yet develop normally to adulthood and are fertile. Detailed histological analysis revealed normal development of the genitourinary system. PMID- 12640140 TI - Native polycystin 2 functions as a plasma membrane Ca2+-permeable cation channel in renal epithelia. AB - Mutations in polycystin 2 (PC2), a Ca(2+)-permeable cation channel, cause autosomal dominant polycystic kidney disease. Whether PC2 functions in the endoplasmic reticulum (ER) or in the plasma membrane has been controversial. Here we generated and characterized a polyclonal antibody against PC2, determined the subcellular localization of both endogenous and transfected PC2 by immunohistochemistry and biotinylation of cell surface proteins, and assessed PC2 channel properties with electrophysiology. Endogenous PC2 was found in the plasma membrane and the primary cilium of mouse inner medullar collecting duct (IMCD) cells and Madin-Darby canine kidney (MDCK) cells, whereas heterologously expressed PC2 showed a predominant ER localization. Patch-clamping of IMCD cells expressing endogenous or heterologous PC2 confirmed the presence of the channel on the plasma membrane. Treatment with chaperone-like factors facilitated the translocation of the PC2 channel to the plasma membrane from intracellular pools. The unitary conductances, channel kinetics, and other characteristics of both endogenously and heterologously expressed PC2 were similar to those described in our previous study in Xenopus laevis oocytes. These results show that PC2 functions as a plasma membrane channel in renal epithelia and suggest that PC2 contributes to Ca(2+) entry and transport of other cations in defined nephron segments in vivo. PMID- 12640142 TI - Disruption of mouse CD46 causes an accelerated spontaneous acrosome reaction in sperm. AB - Human membrane cofactor protein (MCP, CD46) is a ubiquitously expressed protein known to protect cells from complement attack. Interestingly, when we examined the expression of mouse CD46, which we recently cloned, the message was found only in testis and the protein was found on the inner acrosomal membrane of sperm. In order to elucidate the function of CD46, we produced mice carrying a null mutation in the CD46 gene by using homologous recombination. Despite the absence of CD46, the mice were healthy and both sexes were fertile. However, to our surprise, the fertilizing ability of males appeared to be facilitated by disruption of the CD46 gene, as the average number of pups born from CD46(-/-) males was significantly greater than that of wild-type males. It was also revealed that the incidence of the spontaneous acrosome reaction doubled in CD46( /-) sperm compared to that in wild-type sperm. It was assumed that this increase caused the heightened fertilizing ability found in CD46(-/-) sperm. These data suggest that CD46 may have some role in regulating sperm acrosome reaction. PMID- 12640143 TI - Targeted inhibition of Stat3 with a decoy oligonucleotide abrogates head and neck cancer cell growth. AB - The transcription factor signal transducer and activator of transcription 3 (Stat3) is constitutively activated in a variety of cancers including squamous cell carcinoma of the head and neck (SCCHN). Previous investigations have demonstrated that activated Stat3 contributes to a loss of growth control and transformation. To investigate the therapeutic potential of blocking Stat3 in cancer cells, we developed a transcription factor decoy to selectively abrogate activated Stat3. The Stat3 decoy was composed of a 15-mer double-stranded oligonucleotide, which corresponded closely to the Stat3 response element within the c-fos promoter. The Stat3 decoy bound specifically to activated Stat3 and blocked binding of Stat3 to a radiolabeled Stat3 binding element. By contrast, a mutated version of the decoy that differed by only a single base pair did not bind the activated Stat3 protein. Treatment of head and neck cancer cells with the Stat3 decoy inhibited proliferation and Stat3-mediated gene expression, but did not decrease the proliferation of normal oral keratinocytes. Thus, disruption of activated Stat3 by using a transcription factor decoy approach may serve as a novel therapeutic strategy for cancers characterized by constitutive Stat3 activation. PMID- 12640144 TI - Genetic architecture and evolutionary constraint when the environment contains genes. AB - The environment provided by conspecifics is often the most important component of the environment experienced by individuals, frequently having profound effects on fitness and trait expression. Although these social effects on fitness and trait expression may appear to be purely environmental, they differ from other sorts of environmental influences, because they can have a genetic basis and thus can contribute to evolution. Theory has shown that these effects modify the definition of genetic architecture by making the phenotype the property of the genotypes of multiple individuals and alter evolutionary dynamics by introducing additional heritable components contributing to trait evolution. These effects suggest that genetic and evolutionary analyses of traits influenced by social environments must incorporate the genetic components of variation contributed by these environments. However, empirical studies incorporating these effects are generally lacking. In this paper, I quantify the contribution of genetically based environmental effects arising from social interactions during group rearing to the quantitative genetics of body size in Drosophila melanogaster. The results demonstrate that the genetic architecture of body size contains an important component of variation contributed by the social environment, which is hidden to ordinary genetic analyses and opposes the direct effects of genes on body-size development within a population. Using a model of trait evolution, I show that these effects significantly alter evolutionary predictions by providing hidden constraints on phenotypic evolution. The importance of relatedness of interactants and the potential impact of kin selection on the evolution of body size are also examined. PMID- 12640145 TI - Insight into the mechanism of aromatic hydroxylation by toluene 4-monooxygenase by use of specifically deuterated toluene and p-xylene. AB - The present studies address the mechanism of aromatic hydroxylation used by the natural and G103L isoforms of the diiron enzyme toluene 4-monooxygenase. These isoforms have comparable catalytic parameters but distinct regiospecificities for toluene hydroxylation. Hydroxylation of ring-deuterated p-xylene by the natural isoform revealed a substantial inverse isotope effect of 0.735, indicating a change in hybridization from sp(2) to sp(3) for hydroxylation at a carbon atom bearing the deuteron. During the hydroxylation of 4-(2)H(1)- and 3,5-(2)H(2) toluene, similar magnitudes of intramolecular isotope effects and patterns of deuterium retention were observed from both isoforms studied, indicating that the active-site mutation affected substrate orientation but did not influence the mechanism of hydroxylation. The results with deuterated toluenes show inverse intramolecular isotope effects for hydroxylation at the position of deuteration, normal secondary isotope effects for hydroxylation adjacent to the position of deuteration, near-quantitative deuterium retention in m-cresol obtained from 4 (2)H(1)-toluene, and partial loss of deuterium from all phenolic products obtained from 3,5-(2)H(2)-toluene. This combination of results suggests that an active site-directed opening of position-specific transient epoxide intermediates may contribute to the chemical mechanism and the high degree of regiospecificity observed for aromatic hydroxylation in this evolutionarily specialized diiron enzyme. PMID- 12640147 TI - Gastrointestinal stromal tumors: radiologic features with pathologic correlation. AB - Gastrointestinal stromal tumors (GISTs) are unique neoplasms that occur throughout the gastrointestinal tract, mesentery, omentum, and retroperitoneum. They are the most common mesenchymal neoplasm of the gastrointestinal tract and are defined by their expression of KIT (CD117), a tyrosine kinase growth factor receptor. The expression of KIT is important to distinguish GISTs from other mesenchymal neoplasms such as leiomyomas, leiomyosarcomas, schwannomas, and neurofibromas and to determine the appropriateness of KIT-inhibitor therapy. The series described herein was accumulated over 2 years and includes 64 pathologically proved GISTs (28 gastric, 27 small intestinal, six anorectal, one colonic, one esophageal, and one from the small bowel mesentery). Radiologic features of GISTs vary depending on tumor size and organ of origin. Since most GISTs arise within the muscularis propria of the stomach or intestinal wall, they most commonly have an exophytic growth pattern and manifest as dominant masses outside the organ of origin. Dominant intramural and intraluminal masses are less common radiologic manifestations. GISTs occurring in the gastrointestinal tract and mesentery characteristically have hemorrhage, necrosis, or cyst formation that appears as focal areas of low attenuation on computed tomographic images. Although the radiologic features of GISTs are often distinct from those of epithelial tumors, criteria to separate GISTs radiologically from other nonepithelial tumors have not yet been fully developed. PMID- 12640148 TI - Splenic torsion. PMID- 12640146 TI - Histone deacetylase inhibitors prevent oxidative neuronal death independent of expanded polyglutamine repeats via an Sp1-dependent pathway. AB - Oxidative stress is believed to be an important mediator of neurodegeneration. However, the transcriptional pathways induced in neurons by oxidative stress that activate protective gene responses have yet to be fully delineated. We report that the transcription factor Sp1 is acetylated in response to oxidative stress in neurons. Histone deacetylase (HDAC) inhibitors augment Sp1 acetylation, Sp1 DNA binding, and Sp1-dependent gene expression and confer resistance to oxidative stress-induced death in vitro and in vivo. Sp1 activation is necessary for the protective effects of HDAC inhibitors. Together, these results demonstrate that HDAC inhibitors inhibit oxidative death independent of polyglutamine expansions by activating an Sp1-dependent adaptive response. PMID- 12640149 TI - Persistent Mullerian duct syndrome. PMID- 12640150 TI - Clinical role of FDG PET in evaluation of cancer patients. AB - Positron emission tomography (PET) is a diagnostic imaging technique that allows identification of biochemical and physiologic alterations in tumors. Use of PET performed with 2-[fluorine-18]fluoro-2-deoxy-D-glucose (FDG) significantly improves the accuracy of tumor imaging. In terms of oncologic applications, FDG PET has already gained widespread acceptance in the initial staging of cancer, management of recurrent cancer, and monitoring the response to therapy. With conventional imaging modalities, size criteria are used to distinguish between benign and malignant disease in lymph nodes; conversely, FDG PET is based on identification of fundamental aspects of tumor metabolism. FDG uptake in tumors is proportional to the metabolic rate of viable tumor cells, which have an increased demand for glucose. The high sensitivity and high negative predictive value of FDG PET in most malignant tumors enable this technique to play an even greater role in tumor management at initial staging and follow-up. PMID- 12640151 TI - Radionuclide bone imaging: an illustrative review. AB - Bone scintigraphy with technetium-99m-labeled diphosphonates is one of the most frequently performed of all radionuclide procedures. Radionuclide bone imaging is not specific, but its excellent sensitivity makes it useful in screening for many pathologic conditions. Moreover, some conditions that are not clearly depicted on anatomic images can be diagnosed with bone scintigraphy. Bone metastases usually appear as multiple foci of increased activity, although they occasionally manifest as areas of decreased uptake. Traumatic processes can often be detected, even when radiographic findings are negative. Most fractures are scintigraphically detectable within 24 hours, although in elderly patients with osteopenia, further imaging at a later time is sometimes indicated. Athletic individuals are prone to musculoskeletal trauma, and radionuclide bone imaging is useful for identifying pathologic conditions such as plantar fasciitis, stress fractures, "shin splints," and spondylolysis, for which radiographs may be nondiagnostic. A combination of focal hyperperfusion, focal hyperemia, and focally increased bone uptake is virtually diagnostic for osteomyelitis in patients with nonviolated bone. Bone scintigraphy is also useful for evaluating disease extent in Paget disease and for localizing avascular necrosis in patients with negative radiographs. Radionuclide bone imaging will likely remain a popular and important imaging modality for years to come. PMID- 12640152 TI - Imaging of complications of acute mastoiditis in children. AB - Acute mastoiditis is a serious complication of acute otitis media in children. Suppurative disease in the mastoid region occasionally spreads to the adjacent dura mater of the posterior and middle cranial fossae and the sigmoid sinus by means of thrombophlebitis, osseous erosion, or anatomic pathways, producing intracranial complications. Computed tomography (CT) should be performed early in the course of the disease to classify the mastoiditis as incipient or coalescent and to detect intracranial complications. On the basis of the clinical features and imaging findings, the disease is managed conservatively with intravenously administered antibiotics or treated with mastoidectomy and drainage plus antibiotic therapy. CT is therefore a decisive diagnostic tool in determining the type of therapy. In addition, magnetic resonance imaging is performed in patients with clinical symptoms or CT findings suggestive of intracranial complications because of its higher sensitivity for detection of extraaxial fluid collections and associated vascular problems. PMID- 12640153 TI - Aids to successful shoulder arthrography performed with a fluoroscopically guided anterior approach. AB - One method for performing effective shoulder arthrography with a fluoroscopically guided anterior approach is described. The technique can ensure success while injury to the cartilaginous labrum is avoided, which is essential when arthrography is performed in conjunction with magnetic resonance (MR) imaging. The key features of the technique include supine positioning of the patient with the shoulder in external rotation, marking the skin just lateral to the humeral head cortex, inserting the needle perpendicular to the fluoroscopy beam, testing the injection with an anesthetic agent, and confirming intraarticular needle placement with iodinated contrast material. The procedure can be followed by an injection of dilute gadolinium solution for subsequent MR imaging. Proper patient and needle positioning as well as accurate confirmation of intraarticular needle placement are critical to a successful and atraumatic shoulder arthrographic examination. PMID- 12640155 TI - US of the shoulder: non-rotator cuff disorders. AB - The most common indication for shoulder ultrasonography (US) is the diagnosis of rotator cuff disease. However, there is a spectrum of non-rotator cuff abnormalities that are amenable to US examination, including instability of the biceps tendon, glenohumeral joint, and acromioclavicular joint; arthropathies and bursites (inflammatory diseases, degenerative and infiltrative disorders, infections); nerve entrapment syndromes; and space-occupying lesions. Many of these conditions may be overlooked clinically or can even mimic rotator cuff tears, and US can help redirect the diagnosis if a complete shoulder examination rather than a simple rotator cuff assessment is performed. In addition, US can be remarkably helpful in guiding either needle aspiration procedures or local injection therapy in patients with synovial processes. Although radiography, magnetic resonance (MR) imaging, and computed tomographic and MR arthrography are effective modalities for the evaluation of non-rotator cuff disorders, US is both less costly and less invasive and will likely be used more frequently in this setting as experience increases. Once adequate radiographs have been obtained to exclude apparent bone disorders, high-resolution US should be the first-line imaging modality in the assessment of non-rotator cuff disorders of the shoulder, assuming the study is performed with high-end equipment by an experienced examiner. PMID- 12640156 TI - MR imaging of common and uncommon large pelvic masses. AB - Magnetic resonance (MR) imaging is often used in the detection and staging of large pelvic masses. Many large masses in the female pelvis arise from the reproductive organs (eg, uterus, cervix, ovaries, fallopian tubes). In addition, these masses may arise from the gastrointestinal system, urinary system, adjacent soft tissues, peritoneum, or retroperitoneum or from metastases. The majority of large masses in the female pelvis represent such commonly encountered entities as uterine fibroid tumor, dermoid tumor, ovarian cyst, and ovarian cancer. However, uncommon pelvic masses such as mesothelioma, adenocarcinoma, carcinosarcoma, leiomyosarcoma, and desmoid tumor may also be seen. Thus, the differential diagnosis for female pelvic masses is extensive. However, the site of origin, MR imaging characteristics, and clinical history may all help narrow the differential diagnosis. Although with large tumors it may not always be possible to determine the site of origin or distinguish between various tumors at radiology, familiarity with the clinicopathologic and MR imaging features of common and uncommon pelvic masses is important for diagnosis and treatment. PMID- 12640157 TI - MR imaging of the uterine cervix: imaging-pathologic correlation. AB - Magnetic resonance (MR) imaging is useful not only for preoperative staging of gynecologic malignancies but also for prediction of the histopathologic features of a variety of intrapelvic tumors. Familiarity with the specific imaging findings that have been reported for the uterine cervix is a goal of radiologists. The typical MR imaging findings of uterine cervical lesions correspond to the histopathologic features. These lesions can be categorized as epithelial neoplasms, nonepithelial neoplasms, and nonneoplastic diseases. Cervical carcinoma accounts for most cases of malignant lesions and is staged by using the classification system established by the International Federation of Gynecology and Obstetrics. MR imaging allows differentiation between endophytic and exophytic growth and between normal and abnormal findings after hysterectomy and irradiation. Other epithelial neoplasms of the uterine cervix include adenoma malignum, which is a special type of cervical adenocarcinoma, as well as carcinoid tumor and malignant melanoma. Nonepithelial neoplasms of the uterine cervix include malignant lymphoma and leiomyoma. Nonneoplastic diseases of the uterine cervix include cervical pregnancy, cervicitis, nabothian cysts, polyps, and endometriosis. PMID- 12640158 TI - Papillary neoplasms of the bile duct that mimic biliary stone disease. AB - Papillary tumors of the bile duct are intraductal tumors with innumerable minute, frondlike papillary projections. These tumors may be either fixed to or detached from the bile duct wall. However, because the papillary projections on the surface of papillary tumors are long and slender, the tumors are friable and slough easily. The sloughed tumor fragments may float within the bile ducts, resulting in intermittent partial biliary obstruction and mimicking bile duct stones at clinical examination and at ultrasonography (US), computed tomography (CT), and cholangiography. A tumor manifests radiologically as thickening and irregularity of the bile duct wall or as a fixed or sloughed intraductal mass. A nonshadowing intraductal echogenic cast seen at US, an intraductal noncalcified soft-tissue mass with asymmetric wall thickening seen at CT, and an intraductal mass with a papillary surface and a serrated bile duct margin seen at cholangiography are all appearances that suggest a papillary tumor and may be helpful in differentiating a tumor from a bile duct stone. PMID- 12640160 TI - Mesenteric neoplasms: CT appearances of primary and secondary tumors and differential diagnosis. AB - Computed tomography (CT) remains the optimal imaging modality for diagnosing tumors in the mesentery. Although primary neoplasms arising from the mesenchymal tissues of the mesentery are rare, the small bowel mesentery is a major avenue for the dissemination of tumor within the peritoneal cavity. Tumors spread to the mesentery by four major routes: (a) direct extension, commonly seen with carcinoid tumor of the small intestine as well as intraabdominal cancers such as pancreatic and colon cancer; (b) lymphatic dissemination of lymphoma and some epithelial malignancies; (c) hematogenic spread resulting in embolic metastases to the small intestinal wall, usually seen in melanoma and breast cancer; and (d) seeding through the peritoneum from ovarian and gastrointestinal malignancies as well as some lymphomas. Although percutaneous imaging-guided or surgical biopsy is often necessary to guide management, analysis of CT features along with the clinical history may be useful in differentiating mesenteric tumors from infectious, inflammatory, or vascular processes affecting the mesentery. The article presents the characteristic appearances of primary and secondary mesenteric neoplasms at CT and offers a rational approach to the differential diagnosis of mesenteric masses depicted at CT. PMID- 12640161 TI - Hydatid disease from head to toe. AB - Hydatid disease (HD) is a unique parasitic disease that is endemic in many parts of the world. HD can occur almost anywhere in the body and demonstrates a variety of imaging features that vary according to growth stage, associated complications, and affected tissue. Radiologic findings range from purely cystic lesions to a completely solid appearance. Calcification is more common in HD of the liver, spleen, and kidney. HD can become quite large in compressible organs. Hydatid cysts (HCs) can be solitary or multiple. Chest radiography, ultrasonography (US), computed tomography (CT), magnetic resonance (MR) imaging, and even urography can depict HCs. The imaging method used depends on the involved organ and the growth stage of the cyst. US most clearly demonstrates the hydatid sands in purely cystic lesions, as well as floating membranes, daughter cysts, and vesicles. CT is best for detecting calcification and revealing the internal cystic structure posterior to calcification. MR imaging is especially helpful in detecting HCs of the central nervous system. Radiologic and serologic findings can generally help establish the diagnosis of HD, but an HC in an unusual location with atypical imaging findings may complicate the differential diagnosis. Nevertheless, familiarity with imaging findings, especially in patients living in endemic regions, is advantageous in this context. PMID- 12640162 TI - Color Doppler imaging of posttraumatic priapism before and after selective embolization. AB - High-flow priapism usually follows perineal or penile trauma with disruption of an intracavernosal artery. Angiographic embolization of the lacerated artery is currently considered the treatment of choice. The contribution of gray-scale and color Doppler ultrasonography (US) in diagnosis and treatment of 10 patients with high-flow priapism was investigated. In patients with recent arterial laceration, the cavernous tissue surrounding the arterial-sinusoidal fistula appears as a hypoechoic region with undefined margins. In long-standing priapism, this area is usually more regular and circumscribed, mimicking a pseudoaneurysm. Color Doppler US is highly sensitive for detection of the arterial-sinusoidal fistula that causes extravasation of blood from the lacerated cavernosal artery. After angiography, color Doppler US allows confirmation of both successful embolization by demonstrating disappearance or size reduction of the fistula and unsuccessful treatment by demonstrating patency of collateral feeding vessels or early recanalization of the embolized artery. Limitations of color Doppler US include underestimation of the number of accessory feeding vessels, which may become patent only after embolization of the main vascular supply, and difficulty in recognizing vessels that feed the fistula from the opposite side. PMID- 12640163 TI - MR imaging: a "one-stop shop" modality for preoperative evaluation of potential living kidney donors. AB - At many institutions, magnetic resonance (MR) angiography is the technique of choice for assessment of the renal arteries and renal parenchyma in potential living kidney donors. The renal arteries and renal veins have a varied anatomy and may consist of one or more vessels at several levels with variable calibers and levels of branching. These findings may play an important role in the surgeon's decision about which kidney to harvest, especially if laparoscopic nephrectomy is used. A comprehensive MR imaging protocol is used at one hospital to assess the arteries, veins, parenchyma, and collecting system of the kidneys. The protocol includes T2-weighted single-shot fast spin-echo imaging, fat saturated T2-weighted fast spin-echo imaging, three-dimensional MR angiography and MR venography, and delayed fat-saturated three-dimensional T1-weighted gradient-echo imaging. Meticulous assessment of the source images as well as images produced with various postprocessing methods, such as full maximum intensity projection, targeted maximum intensity projection, and axial and oblique reformation, allows detailed description of the vascular anatomy and its relationship to the collecting system and parenchyma to facilitate the surgeon's decision making. The findings of MR imaging are comparable with those of other imaging modalities. PMID- 12640165 TI - Using the IHE scheduled work flow integration profile to drive modality efficiency. AB - The IHE (Integrating the Healthcare Enterprise) Scheduled Work Flow integration profile describes a communication and work flow environment that provides benefits for radiology departments who want to standardize system software. The IHE technical framework defines this environment by specifying the systems involved and the messages exchanged by those systems. The Modality Worklist is a key component of the Scheduled Work Flow integration profile that allows an operator at each modality in the department to retrieve a list of scheduled procedure steps to perform and to automate the process of entering the correct patient identification information in all the images created with the modality. The IHE technical framework defines two transactions used by the modality to tell the Image Manager and Order Filler what was performed and how many images were acquired: the Modality Procedure Step in Progress and Modality Procedure Step Completed. Users who specify the Scheduled Work Flow integration profile will benefit immediately by achieving a certain baseline of functionality. However, users will derive further benefits of increased operational efficiency through negotiation with the providers of software solutions. The integration profile defines features that are optional; users should evaluate these features and request those that are determined to be beneficial. PMID- 12640171 TI - Timing is everything--delaying therapy for delayed cell death. PMID- 12640172 TI - New approaches to brain injury in preterm infants. PMID- 12640173 TI - Under what circumstances can seizures produce hippocampal injury: evidence for age-specific effects. AB - Mesial temporal sclerosis (MTS) is the characteristic hippocampal pathology of temporal lobe epilepsy in adults. Both clinical and experimental studies indicate that although the immature brain is highly susceptible to seizures, it is more resistant to the development of the seizure-induced hippocampal pathology akin to MTS, compared with the adult brain. However, seizures in the immature brain may produce age-specific effects on hippocampal morphology or function. The spectrum of these effects is still unknown. Factors such as the presence of prior neurological abnormalities, age, etiology of the seizures, repetitive seizures and genetic predisposition may affect the range and severity of hippocampal changes. The key point is to identify the significance of these changes and design age-appropriate preventative treatments. PMID- 12640174 TI - Animal models of developmental brain injury: relevance to human disease. A summary of the panel discussion from the Third Hershey Conference on Developmental Cerebral Blood Flow and Metabolism. PMID- 12640175 TI - Prolonged neonatal seizures exacerbate hypoxic-ischemic brain damage: correlation with cerebral energy metabolism and excitatory amino acid release. AB - BACKGROUND: Perinatal hypoxia-ischemia (HI) is the most common precipitant of seizures in the first 24-48 h of a newborn's life. In a previous study, our laboratory developed a model of prolonged, continuous electrographic seizures in 10-day-old rat pups using kainic acid (KA) as a proconvulsant. Groups of animals included those receiving only KA, or HI for 15 or 30 min, followed by KA infusion. Our results showed that prolonged electrographic seizures following 30 min of HI resulted in a marked exacerbation of brain damage. We have undertaken studies to determine alterations in hippocampal high-energy phosphate reserves and the extracellular release of hippocampal amino acids in an attempt to ascertain the underlying mechanisms responsible for the damage promoted by the combination of HI and KA seizures. METHODS: All studies were performed on 10-day old rats. Five groups were identified: (1) group I--KA alone, (2) group II--15 min of HI plus KA, (3) group III--15 min of HI alone, (4) group IV--30 min of HI plus KA, and (5) group VI--30 min of HI alone. HI was induced by right common carotid artery ligation and exposure to 8% oxygen/balance nitrogen. Glycolytic intermediates and high-energy phosphates were measured. Prior to treatment, at the end of HI (both 15 and 30 min), prior to KA injection, and at 1 (onset of seizures), 3, 5 (end of seizures), 7, 24 and 48 h, blood samples were taken for glucose, lactate and beta-hydroxybutyrate. At the same time points, animals were sacrificed by decapitation and brains were rapidly frozen for subsequent dissection of the hippocampus and measurement of glucose, lactate, beta hydroxybutyrate, adenosine triphosphate (ATP) and phosphocreatine (PCr). In separate groups of rats as defined above, microdialysis probes (CMA) were stereotactically implanted into the CA2-3 region of the ipsilateral hippocampus for measurement of extracellular amino acid release. Dialysate was collected prior to any treatment, at the end of HI (15 and 30 min), prior to KA injection, and at 1 (onset of seizures), 3, 5 (end of seizures), 7 and 9 h. Determination of glutamate, serine, glutamine, glycine, taurine, alanine, and GABA was accomplished using high-performance liquid chromatography with EC detection. RESULTS: Blood and hippocampal glucose concentrations in all groups receiving KA were significantly lower than control during seizures (p < 0.05). beta Hydroxybutyrate values displayed the inverse, in that values were significantly higher (p < 0.01) in all KA groups compared with pretreatment controls during seizure activity. Values returned to control by 2 h following the cessation of seizures. Lactate concentrations in brain and blood mimicked those of beta hydroxybutyrate. ATP values declined to 0.36 mmol/l in both the 15 and 30 min hypoxia groups compared with 1.85 mmol/l for controls (p < 0.01). During seizures, ATP and PCr values declined significantly below their homologous controls. Following seizures, ATP values only for those animals receiving KA plus HI for 30 min remained below their homologous controls for at least 24 h. Determination of amino acid release revealed elevations of glutamate, glycine, taurine, alanine and GABA above pretreatment control during HI, with a return to normal prior to KA injections. During seizures and for the 4 h of recovery monitored, only glutamate in the combined HI and KA group rose significantly above both the 15 min of HI plus KA and the KA alone group (p < 0.05). CONCLUSION: Under circumstances in which there is a protracted depletion of high energy phosphate reserves, as occurs with a combination of HI- and KA-induced seizures, excess amounts of glutamate become toxic to the brain. The latter may account for the exacerbation of damage to the newborn hippocampus, and serve as a target for future therapeutic intervention. PMID- 12640176 TI - Neuroprotection of creatine supplementation in neonatal rats with transient cerebral hypoxia-ischemia. AB - We hypothesized that creatine (Cr) supplementation would preserve energy metabolism and thus ameliorate the energy failure and the extent of brain edema seen after severe but transient cerebral hypoxia-ischemia (HI) in the neonatal rat model. Six-day-old (P6) rats received subcutaneous Cr monohydrate injections for 3 consecutive days (3 g/kg body weight/day), followed by 31P-magnetic resonance spectroscopy (MRS) at P9. In a second group, P4 rats received the same Cr dose as above for 3 days prior to unilateral common carotid artery ligation followed 1 h later by 100 min of hypoxia (8% O2) at P7. Rats were maintained at 37 degrees C rectal temperature until magnetic resonance imaging was performed 24 h after HI. Cr supplementation for 3 days significantly increased the energy potential, i.e. the ratio of phosphocreatine to beta-nucleotide triphosphate (PCr/betaNTP) and PCr/inorganic phosphate (PCr/Pi) as measured by 31P-MRS. Rats with hemispheric cerebral hypoxic-ischemic insult that had received Cr showed a significant reduction (25%) of the volume of edemic brain tissue compared with controls as calculated from diffusion-weighted images (DWI). Thus, prophylactic Cr supplementation demonstrated a significant neuroprotective effect 24 h after transient cerebral HI. We hypothesize that neuroprotection is probably due to the availability of a larger metabolic substrate pool leading to a reduction of the secondary energy failure because DWI has been reported to correlate with the PCr/Pi ratio in the acute phase of injury. Additional protection by Cr may be related to prevention of calcium overload, prevention of mitochondrial permeability transition pore opening and direct antioxidant effects. PMID- 12640177 TI - Inhibition of nNOS and iNOS following hypoxia-ischaemia improves long-term outcome but does not influence the inflammatory response in the neonatal rat brain. AB - In this study, we tested the hypothesis that combined inhibition of nNOS and iNOS will reduce neuronal damage and the inflammatory response induced by perinatal hypoxia-ischaemia (HI). In 12-day-old rats, HI was induced by right carotid artery occlusion followed by 90 min of 8% O2. Immediately upon reoxygenation, the rats were treated with NOS inhibitors (n = 24) or placebo (n = 24). Neuropathology was scored at 6 weeks after HI on a 4-point scale (n = 12 per group). The expression of heat shock protein 70 (HSP70) and mRNA expression for cytokines were measured 12 h after HI (n = 12 per group). Histopathological analysis showed that the ipsilateral hemisphere in the NOS inhibition group was less damaged than in the placebo group (p < 0.05). HI induced a significant increase in HSP70 levels (p < 0.05) in the ipsilateral hemispheres, which tended to be lower in the NOS inhibition group (p = 0.07). HI induced an increase in mRNA expression for IL-1beta, TNF-alpha and TNF-beta, but there was no difference between the ipsi- and contralateral hemispheres. Combined inhibition of nNOS and iNOS did not induce any change in cytokine expression. We conclude that the long term neuroprotective effects of combined nNOS and iNOS inhibition were not achieved by an altered cytokine response. PMID- 12640178 TI - Effects of selective nitric oxide synthase inhibition on IGF-1, caspases and cytokines in a newborn piglet model of perinatal hypoxia-ischaemia. AB - Selective inhibition of neuronal and inducible nitric oxide synthase (NOS) with 2 iminobiotin previously showed a reduction in brain cell injury. In the present study, we investigated the effects of 2-iminobiotin treatment on insulin-like growth factor-1 (IGF-1) expression, caspase activity and cytokine expression in a newborn piglet model of perinatal hypoxia-ischaemia. Newborn piglets were subjected to 1 h of hypoxia-ischaemia and were treated intravenously with vehicle or 2-iminobiotin. Vehicle-treated piglets showed reduced IGF-1 mRNA expression and increased caspase-3 activity and DNA fragmentation. 2-Iminobiotin treatment, administered immediately upon reperfusion, prevented these observations. No differences in caspase-8 and -9 activity and cytokine [interleukin (IL) 1alpha/beta, IL-6, tumour necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta] mRNA expression were demonstrated between vehicle- and 2-iminobiotin treated piglets at 24 h following hypoxia-ischaemia. IGF-1 mRNA correlated inversely with caspase-3 and transferase-mediated dUTP-biotin in situ nick end labelling score in the cortex, but positively with caspase-8. Cytokine mRNA did not correlate with IGF-1 mRNA, caspase-3 activity or DNA fragmentation. The present results indicate that the previously demonstrated neuroprotective effect of 2-iminobiotin treatment after perinatal hypoxia-ischaemia coincided with a preservation of the endogenous IGF-1 production and reduced caspase-3 activity, but not with a significant decrease in cytokine production. PMID- 12640179 TI - Evidence that p38 mitogen-activated protein kinase contributes to neonatal hypoxic-ischemic brain injury. AB - We tested the response of stress-activated mitogen-activated protein kinases (MAPKs) - p38 MAPK and c-JUN NH2-terminal kinase (JNK) - following hypoxia ischemia (H-I) induced by unilateral carotid artery ligation and hypoxia (8% O2 and 92% N2) for 2.5 h in postnatal-day-7 rats. Phosphorylation of p38 MAPK increased in the hippocampus and cortex immediately following H-I and returned to a basal level 6 h later. In contrast to p38 MAPK, phosphorylation of JNK decreased in the hippocampus and cortex immediately following H-I. Intracerebroventricular administration of two different p38 MAPK inhibitors prior to H-I significantly protected the neonatal brain from H-I injury. Interestingly, p38 MAPK inhibitors did not attenuate caspase-3 activation 24 h after H-I. Thus, these data suggest that p38 MAPKs contribute to the rapid, early component of brain injury following neonatal H-I. PMID- 12640180 TI - Hypoxic preconditioning increases brain glycogen and delays energy depletion from hypoxia-ischemia in the immature rat. AB - Recent studies have shown a protection from cerebral hypoxic-ischemic (HI) brain damage in the immature rat following a prior systemic hypoxic exposure when compared with those not exposed previously. To investigate the mechanism(s) of hypoxic preconditioning, brain glycogen and high-energy phosphate reserves were measured in naive and preconditioned rat pups subjected to HI. Groups in this study included untouched (naive) controls, preconditioned controls (i.e., hypoxia only), preconditioned with HI insult, and naive pups with HI insult. Hypoxic preconditioning was achieved in postnatal-day-6 rats subjected to 8% systemic hypoxia for 2.5 h at 37 degrees C. Twenty-four hours later, they were subjected to unilateral common carotid artery ligation and systemic hypoxia with 8% oxygen at 37 degrees C for 90 min. Animals were allowed to recover from HI for up to 24 h. At specific intervals, animals in each group were frozen in liquid nitrogen for determination of cerebral metabolites. Preconditioned animals showed a significant increase in brain glycogen 24 h following the initial hypoxic exposure, corresponding to the beginning of the HI insult. Measurement at the end of 90 min of HI showed a depletion of high-energy phosphates, ATP and phosphocreatine, in all animals although ATP remained significantly higher in the preconditioned animals. Thus, the energy from increased glycogen following preconditioning slowed high-energy phosphate depletion during HI, thereby allowing for long-term protection. PMID- 12640181 TI - Early appearance of functional deficits after neonatal excitotoxic and hypoxic ischemic injury: fragile recovery after development and role of the NMDA receptor. AB - We sought to determine whether neonatal rats that sustain unilateral cerebral hypoxic-ischemic or excitotoxic insults (1) manifest contralateral sensorimotor deficits during development or in adulthood and (2) recover from those deficits. Seven-day-old (P7) rats received a right intrastriatal injection of the glutamate analog N-methyl-D-aspartate (NMDA). Unilateral hypoxia-ischemia (HI) was induced by right carotid ligation followed by 1.5 h in 8% O2. Both procedures produce neuronal loss in the striatum and sensorimotor cortex. Nonlesioned controls were included. We scored percent forepaw placement on the edge of a horizontal surface, with lateral vibrissa stimulation, from P9 to P19, and at P33 and P50. Then, on P50, rats were treated with the NMDA antagonist MK-801 to determine whether deficits could be reinstated. NMDA- and HI-lesioned rats exhibited a deficit in contralateral vibrissa-stimulated forepaw placing that emerged during the second week of life. Yet, by P33 and P50, the lesioned groups and controls were indistinguishable. MK-801 injection on P50 resulted in transient reinstatement of the placing deficit. After unilateral neonatal excitotoxic or HI brain injury, contralateral sensorimotor deficits are detected, but in many animals, these deficits have resolved by adulthood. Thus, timing of sensorimotor tests may influence their sensitivity for detection of focal neuropathology originating in the neonatal period. PMID- 12640182 TI - Damage to the choroid plexus, ependyma and subependyma as a consequence of perinatal hypoxia/ischemia. AB - Cerebral hypoxia/ischemia (H/I) of the premature infant is a major cause of cerebral palsy and mental retardation. An important determinant of the ultimate outcome from this insult is the extent to which the stem cells and progenitors in the brain are affected. Irreversible injury to these cells will impair normal development of the infant's brain and, hence, its function. In the present study, we examine early intervals after H/I to identify which cells in the periventricular region are most vulnerable. At 0 h of recovery from a perinatal H/I insult, the choroid plexus shows extensive necrotic damage. The adjacent ependymal and subependymal cells are also affected. Swelling of the ependymal and medial subependymal cells is observed; however, these cells rarely sustain permanent damage. By contrast, cells in the most lateral aspect of the subventricular zone (SVZ) show more delayed, but extensive apoptotic and hybrid cell deaths. Interestingly, activated macrophages/microglia are observed adjacent to the swollen ependymal cells as well as within the affected subependyma. We conclude that the choroid plexus is an especially vulnerable structure in the immature brain, whereas the ependymal and adjacent subependymal cells are relatively resistant to damage. As the medial aspect of the SVZ contains neural stem cells, we predict that neural stem cells will be especially resistant to perinatal H/I brain damage. PMID- 12640183 TI - The clinical epidemiology of Hodgkin lymphoma in HIV-infected patients in the highly active antiretroviral therapy (HAART) era. AB - Studies conducted before the introduction of highly active antiretroviral therapy (HAART) suggested that the risk of Hodgkin lymphoma in persons with human immunodeficiency virus (HIV) infection was increased. However, little is known about the features of this malignancy in patients receiving HAART. From January 1996 through December 2001, 23 cases of Hodgkin lymphoma were diagnosed among 3,945 HIV infected patients attending the Harris County Hospital District in Houston, Texas. Twenty (87%) of the HIV-infected patients diagnosed with Hodgkin lymphoma were receiving HAART and 3 (13%) were naive to antiretroviral therapy. The incidence per 1,000 patients of Hodgkin lymphoma in patients receiving HAART was 6.5. The median duration of HAART before the diagnosis of Hodgkin lymphoma was 16 months (range, 7-22 mo). The median CD4 cell count was 235 cells/mm(3) (range, 189-325 cells/mm(3)) for the 20 HIV-infected patients receiving HAART at the time of diagnosis of Hodgkin lymphoma and 90 cells/mm (range, 72-120 cells/mm(3)) for the 3 patients naive for antiretroviral therapy. Among patients with Hodgkin lymphoma receiving HAART, 50% (10/20) had an HIV-RNA viral load in plasma below the level of detection <400 copies/mL). Chemotherapy was administered to all patients, but a complete response was achieved in 30% (6/20) of the patients receiving HAART and 0% (0/3) of the patients naive to antiretroviral therapy. These results suggest that Hodgkin lymphoma has a low incidence in HIV-infected patients receiving HAART, but the malignancy is an aggressive disease with unfavorable clinical outcome in these patients. PMID- 12640184 TI - Peripheral blood lymphocyte subset counts in patients with dermatomyositis: clinical correlations and changes following therapy. AB - Lymphocytopenia has been reported in patients with connective tissue diseases, including dermatomyositis (DM). However, the risk of infectious complications and the changes of lymphocytic subsets during treatment have been poorly investigated in these patients. We investigated the alterations of peripheral blood lymphocyte counts in patients with DM. A retrospective analysis was conducted in patients with an ascertained diagnosis of DM admitted from 1994 to 2000 in both departments of Dermatology of the Saint-Louis Hospital in Paris. All patients had a peripheral blood absolute lymphocyte count available before therapy. From an initial set of 63 patients, 47 were included in the study. The median absolute lymphocyte count was 888/mm(3) (range, 400-4,070). Low peripheral blood CD4+ and CD8+ T-cell and B-cell counts were consistent findings (median CD4+: 382/mm(3); CD8+: 211/mm(3); CD19+: 122/mm(3)). There was a significant increase in lymphocyte count after 1 month (p < 0.0001), 3-6 months (p = 0.001), and 6-12 months (p = 0.0005) of corticosteroid treatment. Infectious events, mainly pneumonia (PCP), occurred in 12 patients. Their initial lymphocyte count was lower than that of patients who did not develop infections (p = 0.0001). These results support the high prevalence of lymphocytopenia in patients with DM and emphasize the risk for opportunistic infections, mainly PCP, in these patients. Further studies are warranted to evaluate the risk/benefit balance of PCP prophylaxis in patients with DM and severe lymphocytopenia. PMID- 12640185 TI - Severe autoimmune cytopenias in treatment-naive hepatitis C virus infection: clinical description of 35 cases. AB - To determine the clinical characteristics and outcome of patients with chronic hepatitis C virus (HCV) infection presenting severe autoimmune cytopenia unrelated to interferon alpha therapy, we analyzed characteristics and outcomes of 35 patients with HCV (16 from our departments and 19 from the literature). We considered active autoimmune hemolytic anemia (AHA) as a decrease of at least 2 g/dL in hemoglobin levels, an increase of at least 0.6 mg/dL in the serum unconjugated bilirubin level, a reticulocyte count >5%, and a positive direct Coombs test. Severe neutropenia was defined as a neutrophil count <0.5 x 10(9)/L, and severe thrombocytopenia as a platelet count <30 x 10(9)/L. We identified the following cytopenias: AHA (17 cases), severe thrombocytopenia (16 cases), aplastic anemia (2 cases), severe neutropenia (1 case), refractory sideroblastic anemia (1 case), and pure red cell aplasia (1 case). Three patients simultaneously presented 2 types of severe cytopenias. Twenty-seven patients (77%) were female and 8 (23%) male, with a mean age at diagnosis of cytopenia of 51.7 years (range, 18-84 yr). Immunologic markers were detected in 19 (68%) of 28 patients, the most frequent being hypocomplementemia in 16 (57%), cryoglobulins in 15 (54%), antinuclear antibodies in 12 (43%), and rheumatoid factor in 5 (18%). Other associated processes were autoimmune diseases in 14 (50%) of 28 and human immunodeficiency virus (HIV) coinfection in 3 (9%) of 32. We found clinical and immunologic differences between HCV patients with AHA and those with severe thrombocytopenia. Patients with HCV-related AHA showed a higher prevalence of associated autoimmune diseases (71%), cryoglobulins (67%), and cirrhosis (59%). All had a good response to corticosteroids, but a poor prognosis (47% mortality). In contrast, patients with HCV-related severe thrombocytopenia had a lower prevalence of associated autoimmune diseases (11%), a poorer response to corticosteroids (55%), and lower mortality (6%), with HIV/HBV coinfections in some patients. The 35 cases presented demonstrate that different types of immune mediated cytopenias may be severe and clinically significant in patients with HCV infection. Hemolytic anemia and severe thrombocytopenia were the most frequent cytopenias observed. Most patients responded well to corticosteroids, although a higher rate of mortality was observed in those with liver cirrhosis. PMID- 12640186 TI - Endogenous endophthalmitis: an 18-year review of culture-positive cases at a tertiary care center. AB - A retrospective chart review of all patients seen at the Cleveland Clinic Foundation with infectious endogenous endophthalmitis between January 1982 and August 2000 revealed 34 affected eyes in 27 patients. During this time, the median incidence of endogenous endophthalmitis was 1.8 cases/year, and 48.1% of patients presented as outpatients. Twenty-six patients presented to an ophthalmologist, and the diagnosis was initially missed in almost half the cases. Eleven patients had an unremarkable physical exam except for eye findings. We found an equal incidence of bacterial and fungal endophthalmitis and a predominance of among the fungal etiologic agents. We did not, however, note a predominance of Gramnegative organisms seen mostly in reports from Asia. The microbiologic diagnosis was based on aqueous and vitreous cultures or positive eye histopathology stains in almost two-thirds of cases. The sensitivity of the Gram stain was poor, but its specificity and positive predictive value were excellent. The vitreous cultures obtained by vitrectomy instruments were more sensitive in making the diagnosis than the vitreous needle biopsy. Aside from blood cultures and eye specimen cultures, half the patients had an additional infectious focus, most frequently a urinary tract infection, whereas infectious endocarditis was seen in a small minority. Twelve patients had visual improvement with treatment with a final visual acuity better than 20/200 in 44% of the eyes. Good visual outcome was associated with visual acuity of 20/200 or better at diagnosis and with the absence of hypopyon. PMID- 12640187 TI - Adrenal involvement in the antiphospholipid syndrome: clinical and immunologic characteristics of 86 patients. AB - To describe the clinical and immunologic characteristics of patients with adrenal involvement and antiphospholipid syndrome (APS), we conducted a computer-assisted (PubMed) search of the literature to identify all cases of primary adrenal insufficiency associated with antiphospholipid antibodies published in English, French, and Spanish from 1983 (when APS was first defined) through March 2002. We reviewed 86 patients (80 from the literature plus 6 from our cohort); 55% were male, and the mean age at presentation was 43 +/- 16 years. Sixty-one (71%) patients had primary APS, and 14 (16%) had systemic lupus erythematosus. In 31 (36%) patients, adrenal insufficiency was the first clinical manifestation of APS. Abdominal pain was present in 55% of patients, followed by hypotension (54%), fever (40%), nausea or vomiting (31%), weakness or fatigue (31%), and lethargy or altered mental status (19%). The main finding in imaging techniques was compatible with adrenal hemorrhage (59%) and in histopathologic study was a hemorrhagic infarction with vessel thrombosis (55%). Lupus anticoagulant was detected in 97% of patients and the anticardiolipin antibodies titer was positive in 93% of patients. Most patients (95%) were positive for the IgG isotype of anticardiolipin antibodies, whereas 40% were positive for the IgM isotype. Baseline cortisol levels were decreased in 98% of patients, ACTH hormone levels were increased in 96% of patients, and the cosyntropin stimulation test was positive in 100% of patients tested. Steroid replacement therapy was the most frequent treatment (84%), followed by anticoagulation (52%) and aspirin (6%). Thirty-two of 35 (91%) patients with prolonged anticoagulant therapy were in good health with a mean follow-up of 25 months, whereas 25 of the 69 (36%) patients with outcome data available had died. The results of the present review stress the clinical importance of systematic screening for lupus anticoagulant and anticardiolipin antibodies in all cases of adrenal hemorrhage or infarction. An initial screening for hypoadrenalism is mandatory in any antiphospholipid antibody-positive patient who complains of abdominal pain and undue weakness or asthenia. PMID- 12640188 TI - Group B streptococcus (Streptococcus agalactiae) pyogenic arthritis in nonpregnant adults. AB - We analyzed the cases of pyogenic arthritis from group B streptococcus (GBS), or, in nonpregnant adults diagnosed in the Hospital Universitari de Bellvitge, a 1,000-bed tertiary care teaching hospital in Barcelona, Spain, during a 10-year period, and we reviewed the available literature to summarize the experience with this infectious entity. From the database of our institution, which does not attend pediatric, obstetric, or burn patients, we collected all microbiologically proven cases of infectious arthritis seen from January 1992 to December 2001. We excluded patients with infection limited to spine; patients with prosthetic joint infection; patients undergoing articular surgery during the year before diagnosis; and those with tuberculous, brucellar, or fungal arthritis. Of a total of 112 patients identified, GBS was the causative organism in 11 (10%) cases. We reviewed the literature using a MEDLINE search (1972-2001), and found 64 additional cases. Of the 75 patients, 34 (45%) were men and 41 (55%) women, with ages ranging from 20 to 87 years (mean age, 57.9 +/- 14.9 yr); 37 patients (49%) were over 60 years. Sixty-eight percent (51/75) of the patients presented with monoarthritis, while in 32% (24/75) more than 1 joint was involved. The most common location was the knee (36%), followed by the shoulder (25%). In 66% (43/65) of cases, bacteremia was documented. In 64% (47/74) of patients, a systemic predisposing factor for infection was noted; the most common conditions were diabetes mellitus, malignancies, and chronic liver diseases. In 31% (23/75) of patients, a concomitant infectious process due to the same microorganism was found, mainly vertebral osteomyelitis and urinary tract infection. Penicillin was the main antibiotic used after bacterial identification; surgical drainage was performed in 36% (27/75) of cases. The overall mortality rate was 9% (7/75). GBS is now a significant causative agent of pyogenic arthritis in nonpregnant adults. In this population, joint infection by GBS is a disease that mainly affects aged patients with underlying medical illnesses; polyarticular involvement, bacteremia, and the presence of a concomitant infectious process are frequent conditions. The case-fatality rate is substantial. PMID- 12640189 TI - Clinical features and metabolic derangements in acquired generalized lipodystrophy: case reports and review of the literature. AB - We present clinical descriptions, metabolic features, and patterns of body fat loss of 16 patients with acquired generalized lipodystrophy (AGL) seen by us over the last 10 years. In addition, we review 63 cases of AGL reported in the literature. Based on these data, we propose new diagnostic criteria for AGL, the essential criterion being selective loss of body fat from large regions of the body occurring after birth. We also propose a subclassification of AGL into 3 varieties, type 1, the panniculitis variety; type 2, the autoimmune disease variety; and type 3, the idiopathic variety, which affect nearly 25%, 25%, and 50% of patients, respectively. Most of the patients presented in childhood and adolescence. Females were affected approximately 3 times more than males. Subcutaneous fat loss was severe and usually affected the face, trunk, abdomen, and extremities. In some patients, fat loss also involved the palms and soles and intraabdominal region; however, the bone marrow and retroorbital fat were preserved in all patients. Clinically, patients may have voracious appetite, fatigue, and acanthosis nigricans. Hepatomegaly was common, mostly due to hepatic steatosis. Most AGL patients had fasting and/or postprandial hyperinsulinemia, diabetes mellitus, hypertriglyceridemia, and low serum levels of high-density lipoprotein cholesterol, leptin, and adiponectin. Diabetes mellitus and hypertriglyceridemia were less prevalent in the panniculitis variety compared with the idiopathic and autoimmune varieties. The management of AGL includes cosmetic surgery for loss of fat. Severe hypertriglyceridemia should be treated with a very low-fat diet and omega-3 polyunsaturated fatty acid supplementation from fish oils. Management of diabetes is difficult and may necessitate insulin therapy in large doses. Insulin sensitizers such as metformin and thiazolidinediones have been used, although their long-term efficacy and safety remain unknown. Subcutaneous administration of recombinant leptin in AGL patients with hypoleptinemia effectively improves hyperglycemia, hypertriglyceridemia, and hepatic steatosis. Leptin therapy, however, remains investigational. Fibrates alone or in combination with statins may be used to treat hypertriglyceridemia. PMID- 12640190 TI - Long-term outcomes in infants born to HIV-infected women. PMID- 12640191 TI - Mannose-binding lectin in HIV infection: relation to disease progression and highly active antiretroviral therapy. AB - To gain insight into the possible role of mannose-binding lectin (MBL) in HIV infection, we analyzed serum levels and the functional complement activation capacity of MBL in different clinical stages of HIV infection during cross sectional analysis (n = 62) and longitudinal testing (n = 23) as well as during highly active antiretroviral therapy (HAART) (n = 40). The results were correlated with serum levels of C-reactive protein (CRP). Our main findings were as follows. MBL levels and the capacity of complement activation by the MBL pathway were increased in HIV-infected patients with advanced clinical disease as shown in both cross-sectional analysis and longitudinal testing. There was no "normalization" of these parameters during HAART. In fact, MBL levels increased during therapy, and this increase was associated with a good virologic response. Although both MBL and CRP are regarded as acute-phase proteins, no correlation was seen between these proteins. Thus, the notably diverse patterns of MBL responses among patients with different clinical courses and treatments suggest that MBL and complement activation by the MBL pathway could be involved in the pathophysiology of HIV infection. It is not inconceivable that the net effects of MBL responses may vary in different clinical settings. PMID- 12640192 TI - Simian immunodeficiency viruses from multiple lineages infect human macrophages: implications for cross-species transmission. AB - Zoonotic transfer of simian immunodeficiency virus (SIV) from chimpanzees and sooty mangabeys to humans has been documented on at least seven occasions. Several recently identified SIV isolates have also been shown to replicate efficiently in human peripheral blood mononuclear cells (PBMCs) in vitro, indicative of the potential for additional cross-species transmission via T cell infection. Although SIV predominantly uses the macrophage-tropic HIV chemokine coreceptor CCR5, little is known about the ability of SIV to infect human macrophages. In this study, 16 SIV isolates belonging to five different primate lentivirus lineages were tested for their ability to infect human monocyte derived macrophages (MDMs). Twelve of the viruses were capable of infecting MDMs, and 11 of these were also able to replicate in human PBMCs. The replication capacity of the isolates differed within and between the various families and was dependent on particular donor macrophages. Our results suggest that most simian lentiviruses characterized to date not only have the ability to infect primary human T lymphocytes but also replicate efficiently in macrophages, thereby increasing the potential for cross-species transmission into the human population. Comparative studies using these isolates may facilitate the identification of characteristics that contribute to virus infectivity and pathogenicity. PMID- 12640193 TI - Placenta and cord blood mitochondrial DNA toxicity in HIV-infected women receiving nucleoside reverse transcriptase inhibitors during pregnancy. AB - Recent studies of pregnant women and animal models have raised concerns regarding potentially serious mitochondrial toxicity-related side effects in infants born to mothers who received nucleoside reverse transcriptase inhibitors (NRTIs) during their pregnancy to prevent HIV-1 perinatal transmission. The aim of this study was to assess mitochondrial DNA (mtDNA) content of cord blood and placenta in HIV-infected pregnant women receiving NRTI compared with HIV-negative women, hypothesizing that placenta and cord blood mtDNA copies per cell would be decreased in women on NRTI therapy. Immediately following delivery, placenta and cord blood were obtained from eight HIV-infected pregnant women on NRTIs and five HIV-negative women. Assessment of mtDNA copies per cell was accomplished by quantitative real-time PCR. The mean mtDNA copies per cell from the placenta of the HIV-infected women compared with HIV-negative women was 152 +/- 119 and 880 +/- 136 ( =.0016), respectively. Similarly, the mean mtDNA copies per cell from the cord blood of the HIV-positive women compared with HIV-negative women was 144 +/- 101 and 865 +/- 331 ( =.0026), respectively. There was a statistically significant decrease in mtDNA copies per cell in placenta and cord blood between the HIV-infected women on NRTIs compared with HIV-negative women. Further studies are needed to better understand the morbidity to infants and mothers treated with NRTI to prevent vertical transmission of HIV. PMID- 12640195 TI - Exposure to antiretroviral therapy in utero or early life: the health of uninfected children born to HIV-infected women. AB - Concerns have been raised over possible adverse effects of prophylactic antiretroviral therapy (ART) on the fetus and newborn. We analyzed data relating to uninfected children enrolled in the European Collaborative Study and investigated the association between ART exposure, perinatal problems, and major adverse health events later in life. Median length of follow-up was 2.2 (0-15.9) years. Of the 2414 uninfected children, 687 (28%) were exposed to ART in all three periods (antenatal, intrapartum, and neonatal). Of the 1008 infants exposed to ART at any time, 906 (90%) were exposed antenatally, 840 (83%) neonatally, and 750 (74%) both antenatally and neonatally. ART exposure was not significantly associated with pattern or prevalence of congenital abnormalities or low birth weight. In multivariate analysis, prematurity was associated with exposure to combination therapy without a protease inhibitor (PI) (OR = 2.66; 95% CI: 1.52 4.67) and with a PI (OR = 4.14; 95% CI: 2.36-7.23). ART exposure was associated with anemia in early life ( <.001). There was no evidence of an association with clinical manifestations suggestive of mitochondrial abnormalities. The absence of serious adverse events in this large cohort of uninfected children exposed to prophylactic ART in the short to medium term is reassuring. PMID- 12640194 TI - Pharmacokinetics of once-daily saquinavir hard-gelatin capsules and saquinavir soft-gelatin capsules boosted with ritonavir in HIV-1-infected subjects. AB - OBJECTIVE: To investigate the pharmacokinetics of once-daily saquinavir (SQV) hard-gelatin capsule (HGC)/ritonavir (RTV), 1600/100 mg, compared with once-daily SQV soft-gelatin capsule (SGC)/RTV, 1600/100 mg. METHODS: We evaluated 13 randomly selected HIV-1-infected subjects taking once-daily SQV SGC/RTV, 1600/100 mg, plus dual nucleoside reverse transcriptase inhibitors (NRTIs) in this pharmacokinetic (PK) substudy. Subjects took 1 week of SQV HGC/RTV and NRTIs, followed by steady-state SQV PK determinations. Subjects then changed to SQV SGC/RTV and NRTIs for 1 week, followed again by steady-state SQV PK determinations. Area under the plasma concentration versus time curve (AUC), maximum concentration (C(max)), minimum concentration (C(min)), time to C(max), and elimination half-life were calculated. RESULTS: There was no significant difference in AUC values between HGCs and SGCs, with a median (plus interquartile range [IQR]) of 50.0 (42.6-71.5) versus 35.5 (28.0-50.2) mg/L/h, respectively ( =.056). Intersubject variability resulted in 4 of 13 subjects on the SQV SGCs and 2 of 13 subjects on the SQV HGCs having a C(min) below the minimum effective concentration of 0.05 mg/L. CONCLUSION: Once-daily SQV HGCs, 1600 mg, boosted with once-daily RTV, 100 mg, resulted in PK parameters that were similar to those observed with 1600 mg of SQV SGC/100 mg RTV once daily. Once-daily SQV HGC/RTV, 1600/100 mg, may be easier to use in developing countries and may increase access where drug costs can be less, the capsule size is smaller, and the need for refrigeration is lessened. PMID- 12640196 TI - Comparison of clients of a mobile health van and a traditional STD clinic. AB - The objective of this study was to determine if there were any demographic, behavioral, and clinical differences between clients seen aboard a mobile sexually transmitted disease (STD)/HIV clinic compared with those seen in a traditional municipal STD/HIV health clinic for receipt of STD/HIV services. Clients seen in the two different settings were interviewed about demographic characteristics, reasons for their visit, STD history, their HIV/STD risk factors, and the risk factors of their sex partners. Clients in both settings were also offered testing for syphilis, gonorrhea, chlamydia, and HIV. Results suggested that clients seen at the mobile clinic were older, more likely to be injecting drug users themselves and/or to have sex partners who were, or had engaged in prostitution for money or drugs. Over half (54.4%) of the mobile clinic clients sought testing for HIV, and they were far less likely to be seeking care for symptoms of an STD. In contrast, only 7.1% of municipal clinic clients indicated HIV testing as the reason for their visit, whereas nearly two thirds (64.5%) reported symptoms of disease. Two percent of municipal clinic clients and 5.4% of mobile clinic clients had a positive HIV test ( p<.001), and 17.8% of STD clinic clients and 5.6% of mobile van clients had a positive gonorrhea and/or test ( p<.001). These data suggest that a mobile STD/HIV clinic may be an effective strategy to reach individuals at high risk for HIV who are not being served by traditional municipal STD/HIV health clinics. PMID- 12640197 TI - HIV knowledge and risk behaviors among Pakistani and Afghani drug users in Quetta, Pakistan. AB - Situated on the Pakistan-Afghan border, Quetta is home to growing numbers of Afghan refugees. We studied HIV knowledge and risk behaviors among Pakistani and Afghani drug users between July 2001 and November 2001. Of 959 drug users, all were male and the majority used heroin. Most were Pakistani (84.8%), 14.9% were Afghani, and 0.3% were Iranian. Relative to Pakistani drug users, a higher proportion of Afghanis reported no formal education, homelessness, and unemployment ( p <.001). Afghanis were more likely to have used an opiate as their first illicit drug (16% vs. 7%, p <.001), to have ever injected (18.8% vs. 12.3%, p =.04), to report needle sharing (72.2% vs. 48.2%, p =.08), or to report a drug user in their family ( p =.08). None of sexually active Afghanis had ever used a condom compared with 5.0% of the Pakistanis ( p =.01). Only 4.3% of Afghans had ever heard of HIV/AIDS compared with 18.3% of Pakistanis ( p <.001). Extremely low levels of HIV/AIDS awareness and high HIV risk behaviors were evident among drug users in Quetta, among whom Afghanis were especially vulnerable. Interventions to prevent transition to injection, needle exchange, and drug treatment are urgently required to prevent blood-borne infections. PMID- 12640198 TI - Factors associated with the use of highly active antiretroviral therapy in patients newly entering care in an urban clinic. AB - Ethnic minority, female, and drug-using patients may be less likely to receive highly active antiretroviral therapy (HAART), despite its proven benefits. We reviewed the medical records of a consecutive population of 354 patients entering care in 1998 at the Thomas Street Clinic, an academically affiliated, public, HIV specialty clinic in Houston, to determine the factors associated with not receiving HAART as recorded in pharmacy records. Ninety-two patients (26.0%) did not receive HAART during at least 6 months of follow-up. Patients who did not receive HAART were more likely to be women and to have missed more than two physician appointments and were less likely to have a CD4 count <200 cells/microL or a viral load > or = 10 copies/mL. In multivariate logistic analysis, missed appointments (OR = 5.85, p<.0001), female sex (OR = 2.53, =.001), and CD4 count > or = 200 cells/microL (OR = 2.50, p=.001) were independent predictors of not receiving HAART. More than half the patients who never received HAART never returned to the clinic after their first appointment. Among patients new to care, women and those with poor appointment adherence were less likely to receive HAART. Efforts to improve clinic retention and further study of the barriers to HAART use in women are needed. PMID- 12640199 TI - Modulation of infection and type 1 cytokine expression parameters by morphine during in vitro coinfection with human T-cell leukemia virus type I and HIV-1. AB - Infection of injection drug users (IDUs) with the human T-cell leukemia viruses (HTLVs) or HIV is considerably higher than in the non-IDU population. Also, coinfection with HIV-1 and HTLV type I (HTLV-I) occurs more frequently. There is little or no information on the effects of opiates (i.e., morphine) on HTLV infection alone or on coinfection of HTLV-I-infected cells with HIV-1. Therefore, in this report, we analyzed the in vitro effects of morphine on HIV or HTLV infection alone as well as on dual infection with HTLV-I and HIV-1. Morphine decreased the in vitro levels of interferon-gamma (IFN gamma) and IL-2 during single infections, and this effect was reversed by the addition of the opioid antagonist naloxone. In contrast, treatment with morphine resulted in a 31% and 36% increase in IFN gamma and IL-2 levels, respectively, during dual infection. In addition, naloxone had an apparent additive effect on the morphine-associated enhancement of IFN gamma and IL-2 expression in the dual-infection model. Despite the high levels of IFN gamma expression, the viability of the coinfected cells in the presence of morphine was maintained. Importantly, morphine treatment was associated with augmented viral reverse transcription activity in dually infected cultures, apparently to the benefit of HTLV-I. If a similar putative morphine induced advantage for HTLV-I production also occurs during in vivo coinfection, opiates such as morphine could contribute to the observed increased rate of HIV 1/HTLV-I infection in the IDU population in a more direct fashion than was previously believed. PMID- 12640200 TI - Public policy regulating private and public space in gay bathhouses. AB - Public policies prohibiting either public or private space in gay bathhouses vary across cities. New York, San Francisco, Los Angeles, and Chicago all have different policies. The objective of this study was to assess reported risk behavior as an indicator of success of one policy over another. Data are from a telephone survey of a probability sample of men living in the four cities who reported having sex with men. Analyses focused on city differences in behavior of adult men who, in the past year, were sexually active with a male and visited a bathhouse ( n= 827). Respondents reported numbers of sex partners, one-night stands, visits to bathhouses in the past year, and casual sex partners with whom they had engaged in unprotected anal intercourse (UAI) as well as whether they had UAI in a public setting. Among men who visit bathhouses, no significant city differences were observed except that there were city differences in where UAI occurred-i.e., San Francisco men were significantly less likely to report UAI in a public place than were men in other cities. The data suggest that different city policies may affect where, but not whether, UAI occurs. PMID- 12640201 TI - Identifying recent HIV infections using the avidity index and an automated enzyme immunoassay. AB - We evaluated a procedure for identifying recent HIV infections, using sequential serum samples from 47 HIV-positive persons for whom the seroconversion date could be accurately estimated. Each serum sample was divided into two aliquots: one diluted with phosphate-buffered saline and the other diluted with 1 M guanidine. We assayed the aliquots with the automated AxSYM HIV1/2gO test (Abbott Diagnostics Division), without modifying the manufacturer's protocol. We then calculated the avidity index (AI): the ratio of the sample/cutoff value for the guanidine aliquot to that of the phosphate-buffered saline aliquot. We analyzed 216 serum samples: 34 samples were collected within 6 months of seroconversion (recent seroconversions), and 182 were collected after 6 months. The mean AIs, by time from seroconversion, were 0.68 +/- 0.16 (within 6 months) and 0.98 +/- 0.10 (after 6 months) (P < 0.0001). AI of <0.90 correctly identified 88.2% of recent infections but misclassified as recent infections 13.2% of serum samples collected afterward. The probability of an infection being classified as recent and having AI of > or = 0.90 would be 0.7% in a population with 5% recent infections. AI can identify with a certain degree of accuracy recent HIV infections, and being a quantitative index, it provides different levels of sensitivity and specificity, depending on the selected cutoff value. The standard assay procedure is not modified. This test is simple and inexpensive and could be used for surveillance, decision-making in treatment, and prevention. PMID- 12640202 TI - Psychological distress and progression to AIDS in a cohort of injection drug users. AB - OBJECTIVE: We investigated whether distress was independently associated with more rapid progression to AIDS among HIV-infected injection drug users (IDUs). DESIGN: A cohort study of IDUs in Baltimore followed from 1988 through 1999. A total of 451 IDUs met the eligibility criteria of being HIV-seropositive but AIDS free at baseline. METHODS: We used Cox proportional hazards models to investigate progression to AIDS within 2 years of baseline, controlling for CD4 lymphocyte count, HIV-1 viral load, and oral thrush. RESULTS: Of the 451 participants, 76.3% were male and 95.8% were African-American; the median age at enrollment was 34 years. The cumulative incidence of AIDS within 2 years of baseline was 7.1%. In multiple regression analysis, distress was associated with more rapid time to AIDS (adjusted relative hazard [RH] = 2.39; 95% CI: 1.16-4.90). The strongest association was observed among IDUs with the lowest (<200 x 10 /L) CD4 counts (adjusted RH = 4.94; 95% CI: 1.30-18.77). CONCLUSIONS: Psychological distress was independently associated with shorter time to AIDS among HIV-infected IDUs, especially among those with the lowest CD4 cell counts, but was not predictive of mortality in this cohort. Further study of the effects of psychological distress on AIDS progression within this population is warranted. PMID- 12640203 TI - Adherence to antiretroviral therapy and persistence of HIV RNA in semen. AB - CONTEXT: Long-term adherence to antiretrovirals is critical for sustained virologic response to HIV therapy in blood. Although antiretroviral therapy (ART) reduces HIV seminal shedding, little is known about the relationship between adherence to ART and HIV suppression in semen. OBJECTIVE: To determine predictors of seminal HIV RNA suppression after 6 months of ART. DESIGN: Prospective observational cohort of 93 HIV-infected subjects before and after introduction of ART. Seminal HIV RNA was measured at baseline and 1, 2, 3, and 6 months after the introduction of therapy. Adherence to therapy was measured by self-report. SETTING: A large academic HIV reference center in Rio de Janeiro, Brazil. MAIN OUTCOME MEASURE: Detectable HIV RNA in semen. RESULTS: In a multivariate logistic model with undetectable seminal HIV RNA after 6 months of therapy as the outcome variable, adjusting for baseline seminal viral load, both being adherent to therapy (OR = 11.8, < 0.01) and using triple-drug ART (OR = 6.48, = 0.04) were independently associated with seminal HIV RNA suppression. CONCLUSIONS: Inability to adhere to therapy was strongly associated with persistent shedding of HIV RNA in semen. Measures to improve adherence are urgently needed to reduce the sexual spread of potentially drug-resistant HIV among subjects using antiretrovirals. PMID- 12640204 TI - A cluster of human T-cell lymphotropic virus type I-associated myelopathy/tropical spastic paraparesis in Jujuy, Argentina. AB - Compared with other regions in Argentina, greater human T-cell lymphotropic virus type I (HTLV-I) seroprevalence has been reported in Jujuy Province, where it reaches 2.32% in the general population, so that a search for HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) cases deserved to be carried out. Accordingly, a clinically diagnosed and serologically confirmed cluster of cases in 1 man and 10 women, including 2 sisters, is described here. Most patients (9/11) were born in Cochinoca Department, located in an Andes highland area called Puna Jujena, situated at more that 3400 m above sea level. No history of risk factors was disclosed, except for a single transfusion in 1 patient. In contrast to the Andean region of Bolivia, where high HTLV-I seroprevalence is in part attributable to Japanese immigrants, the Jujuy population mainly consists of aborigines, mestizos, and European descendants. Therefore, the long-term presence of virus in Jujuy natives may be taken for granted. Considering the HAM/TSP cluster described here plus previously reported isolated cases in neighboring Salta Province, we speculate that the Puna Jujena region and regions in that vicinity would be a microepidemic focus of disease. To determine the role of possible pathogenic cofactors such as geographic, ethnic, genetic, and cultural features, further pertinent surveys are required in subtropical northwestern Argentina. PMID- 12640205 TI - Age differences in sexual partners and risk of HIV-1 infection in rural Uganda. AB - OBJECTIVES: To assess whether differences in age between sexual partners affect the risk of HIV infection in female adolescents and young adults. METHODS: A total of 6177 ever sexually active women aged 15 to 29 years completed a sociodemographic and sexual behavior questionnaire and provided a blood sample for HIV-1 serology. The age difference between partners was categorized as men 0 to 4 years older (referent group), 5 to 9 years older and 10 or more years older. HIV prevalence and incidence were assessed, and adjusted RR was estimated by multivariate regression. RESULTS: Prevalent HIV-1 infection in female participants increased with older male sexual partners. Among women aged 15 to 19 years, the adjusted risk of HIV infection doubled (RR = 2.04; 95% CI: 1.29-3.22) among those reporting male partners 10 or more years older compared with those with male partners 0 to 4 years older; among women 20 to 24 years of age, the RR was 1.24 (95% CI: 0.96-1.60). The attributable fraction (exposed) of prevalent HIV infection in women aged 15 to 24 years associated with partners 10 or more years older was 9.7% (95% CI: 5.2-14.0). HIV incidence did not increase with differences in age of partners. CONCLUSION: The age difference between young women and their male partners is a significant HIV risk factor, suggesting that high HIV prevalence in younger women is caused, in part, by transmission from older male partners. PMID- 12640206 TI - Virologic, immunologic, and clinical response to highly active antiretroviral therapy: the gender issue revisited. AB - BACKGROUND: Highly active antiretroviral therapy (HAART) has dramatically improved the prognosis for patients with HIV. There is ongoing debate over a potential gender effect on patient outcome after HAART. METHODS: Individuals were from the EuroSIDA cohort, naive to protease inhibitors and nonnucleoside reverse transcriptase inhibitors, and had at least one viral load and CD4 measurement prior to starting HAART. Endpoints were virologic (time to <500 copies/mL, time to rebound [first of two consecutive viral loads >500 copies/mL]), immunologic (time to a 100/mm cell rise in CD4 count) and clinical (time to new AIDS and death). Hazard ratios (HR), derived using Cox regression models, compared female to male rates of achieving endpoints. RESULTS: Of 2547 patients, 20% (511) were female. Significantly more females than males were nonwhite (24% vs. 10%, p <.001). Males were older (median age 39 vs. 35 years, p <.0001), had lower CD4 counts (211 vs. 240/mm, p =.03), higher viral loads (4.6 vs. 4.4 log copies/mL, p <.0001), were more likely to have a history of AIDS (26% vs. 18%, p <.001) and were more likely to be treatment-naive (34% vs. 29%, p =.03). Adjusted HR for association between gender (comparing females with males) and the outcomes studied were as follows: for reaching <500 copies/mL 0.91 (0.81-1.03, p =.17), rebound 1.17 (0.95-1.44, p =.15), for 100 cell CD4 count rise 1.02 (0.88-1.14, p =.99), for progression to new AIDS 1.12 (0.73-1.71, p =.59) and for time to death 1.15 (0.69-1.92, p =.57). CONCLUSIONS: We found no significant evidence of a gender difference in virologic, immunologic, or clinical outcomes after starting HAART. PMID- 12640207 TI - HIV-1 RNA levels, resistance, and drug diffusion in semen versus blood in patients receiving a lopinavir-containing regimen. PMID- 12640208 TI - Increased production of oncostatin-M by lymphomononuclear cells from HIV-1 infected patients with neuroAIDS. PMID- 12640209 TI - Is treatment failure for hepatitis C virus infection in HIV-positive drug users associated with a shift in HCV genotypes? PMID- 12640210 TI - The safety of over-the-counter drugs: some reflections and unanswered questions. PMID- 12640211 TI - The efficacy and safety of olanzapine for the treatment of geriatric psychosis. AB - Elderly psychiatric patients often present with psychotic symptoms that need antipsychotic treatment. Olanzapine is one of the atypical antipsychotics with efficacy for psychotic symptoms and a safer side-effect profile than typical antipsychotics. This study was conducted to assess the efficacy and safety of olanzapine for treatment of geriatric psychosis. The sample population comprised 94 acute-ward patients who were 65 years of age or older. Clinical assessment was conducted at baseline and also at 4 weeks after commencement of olanzapine treatment, with use of the Brief Psychiatric Rating Scale (BPRS) and Clinical Global Impression Improvement (CGI-I) instruments. A 4-week therapeutic evaluation was completed for 80 patients, 73 of whom (91.3%) experienced mild to substantial improvement as determined from the CGI-I. A mean 52.6% reduction from baseline was also determined from the BPRS. The mean daily dosage of olanzapine in the fourth week was 10.1 +/- 5.3 mg/d (range, 2.5-20.0). Higher olanzapine dosages were administered for patients with functional psychoses than for an analogous group with organic mental disorders. Adverse effects were monitored for all 94 patients, the most common of which were somnolence (18.1%), dizziness (18.1%), and weakness of legs or bradykinesia (16.0%). Body weight and fasting triglyceride and sugar levels were significantly elevated after olanzapine treatment (2.2, 39.9, and 8.9% from baseline, respectively). It seems reasonable to suggest that olanzapine is efficacious for geriatric patients with psychosis and that the dosage should be diagnosis-dependent. PMID- 12640212 TI - Role of the smoking-induced cytochrome P450 (CYP)1A2 and polymorphic CYP2D6 in steady-state concentration of olanzapine. AB - This study investigated whether the smokinginducible cytochrome P450 (CYP) 1A2 and the polymorphic CYP2D6 play significant roles in the metabolism of olanzapine and its clinical effects at steady-state treatment. Caffeine and debrisoquine were used as measures of CYP1A2 and CYP2D6, respectively. After drug therapy for 15 days, the effect of olanzapine on the activities of CYP1A2 and CYP2D6 was also examined. Seventeen psychiatric patients (9 men and 8 women) were orally administered olanzapine, at a mean +/- standard deviation (SD) dosage of 10 mg/d for all smokers (n = 8) and 7.5 +/- 2.5 mg/d (range, 5-10 mg) for nonsmokers (n = 9;p <0.01). The plasma concentration-to-dose (C:D) ratio was closely correlated to the CYP1A2 activity ( s = -0.89;p <0.0001). The mean urinary caffeine indexes of nonsmokers and smokers were 17 +/- 8 and 101 +/- 44, respectively, indicating that smoking had induced a sixfold higher CYP1A2 activity (p <0.0001). Likewise, the olanzapine plasma C:D ratio (ng.mL.mg) was about fivefold lower in smokers (7.9 +/- 2.6) than in nonsmokers (1.56 +/- 1.1;p <0.0001). On day 15 of the antipsychotic therapy, the percentage decrease in Brief Psychiatric Rating Scale (BPRS) total score relative to the predosing score (in the drug-free period) was higher for nonsmokers than for smokers (30.4 +/- 10% vs. 12.5 +/- 14%;p <0.01). Six nonsmokers and three smokers experienced side effects with olanzapine. After 15 days of drug treatment, olanzapine had caused significant (p <0.0001) and substantial CYP1A2 inhibition (by 50%) in comparison with predosing values, and such inhibition can contribute to adverse drug interactions. In conclusion, smoking-induced increased CYP1A2 activity significantly diminished plasma olanzapine concentrations and the antipsychotic effect of the drug. The performance of a simple caffeine test may assist in individualization of the olanzapine dosage. PMID- 12640213 TI - The effect of risperidone treatment on superoxide dismutase in schizophrenia. AB - Some reports have shown that schizophrenia is accompanied by the abnormal metabolism of free radicals. The purpose of this study was to investigate the effect of the atypical antipsychotic drug risperidone on blood superoxide dismutase (SOD), a critical enzyme in the detoxification of superoxide radicals, and to explore the relationship between changes in SOD and the therapeutic outcome. Forty-one inpatients with diagnosed schizophrenia (DSM-III-R) were assigned to 12 weeks of treatment with risperidone at a fixed dosage of 6 mg/d after a 2-week washout period. Clinical efficacy was determined with the Positive and Negative Syndrome Scale (PANSS). Blood SOD was assayed by radioimmunoassay (RIA) in schizophrenic patients before and after the 12-week treatment, and the values were compared with those of 50 age-, sex-, and smoking-matched subjects without schizophrenia. Risperidone treatment significantly decreased the initially high blood SOD levels in schizophrenia. There was a significantly positive relationship between the change in SOD at pretreatment and posttreatment and the reduction in the PANSS negative subscore. These findings suggest that risperidone treatment significantly decreased the blood SOD levels of schizophrenic patients, a change which may be associated with the diminishment of symptoms. The limitations of this study are the measurement of SOD levels by RIA rather than biochemical assay; the 2-week washout, which may not be adequate; and the measurement of only SOD enzyme and not the other antioxidant enzymes. PMID- 12640214 TI - Acute dysphoric mania: treatment response to olanzapine versus placebo. AB - A substantial number of patients with mania have significant concomitant depressive features, and they may respond differently to mood stabilizers than patients with pure mania. This post-hoc analysis explored the response characteristics of olanzapine versus placebo in bipolar I manic patients with dysphoric and nondysphoric mania (differentiated by baseline Hamilton Depression Rating Scale [HAM-D] score of >20). Two similar, double-blind, randomized trials comparing olanzapine, 5-20 mg, to placebo were pooled for these analyses (N = 246). Mean changes in Young-Mania Rating Scale (Y-MRS) and HAM-D scores during 3 weeks of treatment were examined. Twenty-eight percent of patients had dysphoric mania (olanzapine, n = 33; placebo, n = 35). Among these patients, olanzapine treated patients had greater improvement within 1 week than did placebo-treated patients on both mania ratings (Y-MRS: -9.7 vs. -3.0 points; = 0.011) and depressive symptom ratings (HAM-D: -9.9 vs. -5.4 points; = 0.025). Among those manic subjects without prominent depressive symptoms (olanzapine, n = 91; placebo, n = 87), mean Y-MRS improvement from baseline to endpoint with olanzapine (-11.5 points) versus placebo (-6.13 points) was comparable to the improvement seen with olanzapine versus placebo in the dysphoric mania subgroup ( = 0.476, test of interaction). In acutely ill manic patients with significant depressive symptoms, olanzapine demonstrated a broad spectrum of efficacy, effectively treating both manic and depressive symptoms. The magnitude of the antimanic response appears similar, regardless of baseline depressive features. Additional experience with putative mood stabilizers and atypical agents in mixed mania should include an exploration of their efficacy in treating both manic and depressive mood symptoms. PMID- 12640215 TI - Initial platelet serotonin (5-HT) transport kinetics predict nortriptyline treatment outcome. AB - According to the hypothesis of initial conditions, drug response may be determined by different initial states of neurotransmitter protein recognition systems. Platelet serotonin (5-HT) transport kinetics were studied as initial conditions predictors of antidepressant response in 24 depressed patients before and after 3 weeks of treatment with nortriptyline (75 mg). The initial affinity of the 5-HT transporter (5-HTT) correctly predicted 71% of the outcome. The pretreatment affinity constant ( Km) correlated (r = 0.61; p < 0.002) with that measured after 3 weeks of treatment (Kapp). Responding patients had a significantly higher initial Km before treatment and a significantly higher Kapp after treatment. Nonresponders had an initial Km significantly lower than that of 24 controls. Nortriptyline plasma levels were not statistically different between response groups. These results are consistent with two previously published observations, which indicate that the initial affinity of the 5-HTT predicted response to fluvoxamine or fluoxetine in the same way. Insofar as all three drugs increase the apparent affinity of the 5-HTT, it appears that a better response is related to those cases where the initial affinity is already higher before treatment. PMID- 12640216 TI - Economic evaluation of antidepressive agents: a systematic critique of experimental and observational studies. AB - The purpose of this study was to examine whether experimental and observational pharmacoeconomic analyses of antidepressant drugs support the choice of one of the selective serotonin reuptake inhibitors or newer antidepressants as first line treatment for patients with major depression. We systematically reviewed economic evaluations of two or more antidepressants completed in clinical practice. A systematic electronic search yielded 38 studies meeting the inclusion criteria, of which 23 were administrative database analyses, 12 were observational studies, and 3 were randomized clinical trials. Experimental data indicated that tricyclic antidepressants are equivalent to selective serotonin reuptake inhibitors in terms of total expenditure. While the database analyses are susceptible to bias and confounding variables, they provided an added dimension based on observations from everyday clinical practice. The majority of these studies failed to show any significant difference. Taken together, available pharmacoeconomic studies indicate that tricyclic drugs and selective serotonin reuptake inhibitors have similar cost effectiveness in the health care systems where these comparisons have been made. PMID- 12640217 TI - Clinical and neurobiological effects of tianeptine and paroxetine in major depression. AB - Selective serotonin reuptake inhibitors (SSRIs) are widely used as effective pharmacological agents to treat depressive disorders. In contrast to the SSRIs, which block the presynaptic serotonin (5-HT) transporter and by this route increase the concentration of serotonin in the synaptic cleft, the antidepressant tianeptine enhances the presynaptic neuronal reuptake of 5-HT and thus decreases serotonergic neurotransmission. Both SSRIs and tianeptine are clinically effective; however, their opposite modes of action challenge the prevailing concepts on the need of enhancement of serotonergic neurotransmission. To better understand the differences between these two opposite pharmacological modes of action, we compared the changes induced by tianeptine and paroxetine on psychopathology, the hypothalamic-pituitary-adrenocortical (HPA) system, and cognitive functions in a double-blind, randomized, controlled trial including 44 depressed inpatients over a period of 42 days. Depressive symptomatology significantly improved in all efficacy measures, with no significant differences between tianeptine and paroxetine. There was a trend toward better response to the SSRI among women. Assessment of the HPA system showed marked hyperactivity before the beginning of treatment, which then normalized in most of the patients, without significant differences between the two antidepressants. Cognitive assessments showed no significant differences between the two drugs investigated. The results of the current study suggest that the initial effect, i.e., enhancement or decrease of 5-HT release, is only indirectly responsible for antidepressant efficacy, and they support the notion that downstream adaptations within and between nerve cells are crucial. The normalization of the HPA system as a common mode of action of different antidepressants seems to be of special interest. PMID- 12640218 TI - The role of cytochrome P450 2C19 activity in flunitrazepam metabolism in vivo. AB - Flunitrazepam, a hypnotic benzodiazepine, is widely prescribed around the world for the treatment of insomnia and as a preanesthetic. In vitro studies have shown that the metabolism of flunitrazepam to desmethylflunitrazepam and 3 hydroxyflunitrazepam is mediated in part by the polymorphic enzyme CYP2C19. The objective was to examine the role of CYP2C19 activity in determining flunitrazepam kinetics in vivo. Sixteen healthy volunteers (14 genotypic extensive metabolizers and 2 poor metabolizers) were recruited who had a wide range of CYP2C19 activity (0.50-28.8), as determined by the omeprazole/ 5 hydroxyomeprazole ratio (OMR) at 3 hours following administration of omeprazole, 20 mg orally. Each subject received flunitrazepam, 1 mg orally. Blood samples were collected immediately before and up to 48 hours after drug administration and were assayed by HPLC for flunitrazepam and its metabolites, 7 aminoflunitrazepam, desmethylflunitrazepam, and 3-hydroxyflunitrazepam. Spearman correlations were determined for OMR and pharmacokinetic parameters. With increasing OMR (decreasing CYP2C19 activity), the ratio of flunitrazepam to both desmethylflunitrazepam and 3-hydroxyflunitrazepam AUCs increased ( r = 0.55, p = 0.03 and r = 0.65, p = 0.01, respectively). However, variation in CYP2C19 activity did not significantly affect the AUCs of flunitrazepam or its metabolites. The authors conclude that CYP2C19 contributes to the metabolism of flunitrazepam to desmethylflunitrazepam and 3-hydroxyflunitrazepam in vivo, but these data suggest that its role is minor and that differences in CYP2C19 activity do not likely substantially influence its clinical effects. PMID- 12640219 TI - Safety and efficacy of switching psychiatric patients from a delayed-release to an extended-release formulation of divalproex sodium. AB - This study evaluated the safety and efficacy of divalproex sodium extended release (ER) when patients were switched from therapy with divalproex sodium delayed-release (DR) to divalproex sodium ER. This open-label, 7-day study included 55 patients with bipolar disorder, major depression, schizophrenia, schizoaffective disorder, Alzheimer's disease, dementia, or intermittent explosive disorder. Baseline plasma valproate concentrations were determined, and patients received their usual morning dose of divalproex sodium DR. At 9:00 p.m. the same day, they received divalproex sodium ER at a dose equal to their total daily dose of divalproex sodium DR. Valproate concentrations were monitored, and efficacy was measured with the Positive and Negative Syndrome Scale (PANSS). Side effects were assessed using the Udvalg for Kliniske Undersogelser (UKU) Side Effect Rating Scale. Valproate concentrations for 52 patients remained within the therapeutic range. Inpatient PANSS scores significantly improved from baseline to final evaluation on all subscales. For the combined inpatient and outpatient populations, a small but statistically significant improvement from baseline to final evaluation was seen for positive, general, and total PANSS subscale scores. At study's end, patients reported a significant decrease in the number and severity of adverse events; 54 of 55 patients elected to continue therapy with once-daily divalproex sodium ER. This study suggests that divalproex sodium ER is at least as effective as the DR formulation for treating patients with psychiatric illness and may be better tolerated. The ER formulation offers the advantage of once-daily dosing, which may help improve compliance. PMID- 12640220 TI - Antidepressant properties of anticonvulsant drugs for bipolar disorder. AB - A growing number of anticonvulsant drugs are receiving attention as possible mood stabilizers. This attention is based mainly on the assumption that the antimanic efficacy of anticonvulsants makes them suitable as mood stabilizers. However, their antidepressant properties have received less scrutiny. In this review, current evidence concerning the acute and prophylactic efficacy of divalproex, carbamazepine, gabapentin, lamotrigine, and topiramate in bipolar depression is evaluated. Clinical outcome data are considered, together with limitations of existing studies and the concept of unmet clinical needs. Findings in placebo controlled trials suggest an acute and prophylactic antidepressant effect with lamotrigine monotherapy and more modest antidepressant benefits with other agents administered as monotherapies. Results of published studies are considered with respect to the conceptualization of mood stabilization as arising from antimanic and antidepressant efficacy in bipolar disorder. PMID- 12640221 TI - Low-dose risperidone as adjunctive therapy for irritable aggression in posttraumatic stress disorder. AB - Increased aggressive behavior can occur in association with posttraumatic stress disorder (PTSD). This study tested the hypothesis that low-dose risperidone reduces aggression and other PTSD-related symptoms in combat veterans. Subjects were male combat veterans with PTSD who scored 20 or higher on cluster D (hyperarousal) of the Patient Checklist for PTSD-Military Version (PCL-M). Subjects were randomly assigned to either risperidone or placebo treatment groups. Drugs were administered over a 6-week treatment period in a double-blind manner. Subjects received either risperidone (0.5 mg/day; n = 7) or matched placebo (n = 8) tablets during the first 2 weeks of the treatment period. The dose of risperidone could then be increased up to 2.0 mg/day on the basis of response. Prerandomization psychotropic regimens were continued. Subjects were evaluated with the PCL-M and the Overt Aggression Scale-Modified for Outpatients (OAS-M). In comparison with placebo treatment, reductions in scores between baseline and the last week of treatment were significantly greater for OAS-M irritability and PCL-M cluster B (intrusive thoughts) subscales and on the PCL-M total scale. These results suggest that low-dose risperidone administration reduces irritability and intrusive thoughts in combat-related PTSD. PMID- 12640222 TI - Concomitant use of antipsychotics and drugs that may prolong the QT interval. AB - Concomitant use of drugs that prolong the QT interval is a risk factor for torsades de pointes, a ventricular arrhythmia associated with sudden death. This study compared the concomitant use of drugs that may prolong the QT interval ("other QT drugs") among two groups of patients: one that took antipsychotics that may prolong the QT interval (the QT antipsychotic group, n = 1,750) and one that used antipsychotics that do not result in QT prolongation (the non-QT antipsychotic group, n = 1,139). Data were pharmacy claim and eligibility information from January 1, 2000, through December 31, 2000, from a research database of a large pharmacy benefit manager. Concomitant use of antipsychotics and other QT drugs was examined for each participant over a 3- to 12-month follow up period. Results showed that 51% of QT antipsychotic group members used other QT drugs concomitantly for at least 1 day in the follow-up period. Logistic regression indicated that there was no significant difference between the QT antipsychotic and non-QT antipsychotic groups with concomitant use of other QT drugs when potential confounders were controlled ( p = 0.6013). Although female sex is a risk factor for drug-induced torsades de pointes, women were more likely to concomitantly use other QT drugs than men in both the QT (56.2% vs. 43.2%; p < 0.001) and non-QT (53.1% vs. 43.0%;p < 0.001) antipsychotic groups. Findings suggest that the use of other QT drugs is not being minimized among patients taking QT antipsychotics. PMID- 12640223 TI - Antidepressive treatment with amitriptyline and paroxetine: effects on saliva cortisol concentrations. AB - With regard to the course of basal human hypothalamus-pituitary-adrenal (HPA) system activity, there is a lack of data for comparing different classes of antidepressants. Ninety-four patients were included in a study comparing standardized treatment with paroxetine (PAROX) and amitriptyline (AMI) after a drug-free period of at least 6 days. Saliva for measurement of cortisol concentrations was obtained daily at 0800, 1600, and 2200 during the 6 days of drug-free washout and 35 days of active treatment. The course of HPA system activity and psychopathology, as assessed by the Hamilton Depression Scale, was compared by means of repeated-measurement analyses of variance (ANOVA-rm). Only AMI responders-not PAROX responders or nonresponders to either antidepressant-had a significant decline in saliva cortisol concentrations. In hypercortisolemically depressed patients, treatment with AMI may be preferable to PAROX in order to lower HPA system activity. PMID- 12640224 TI - Psychic akathisia. PMID- 12640225 TI - An open-label trial of modafinil augmentation in patients with partial response to antidepressant therapy. PMID- 12640226 TI - Plasma cotinine, 3'-hydroxycotinine, and their glucuronides in white and black smokers. PMID- 12640227 TI - Nitrous oxide has no effect in the treatment of alcohol withdrawal syndrome: a double-blind placebo-controlled randomized trial. PMID- 12640229 TI - The ALLHAT study: if you believe in evidence-based medicine, stick to it! PMID- 12640228 TI - Kinematic analysis of the effects of donepezil hydrochloride on hand motor function in patients with Alzheimer dementia. PMID- 12640231 TI - Size isn't everything--ALLHAT in perspective. PMID- 12640230 TI - Treating hypertension: it is not how you start but where you end that matters. PMID- 12640232 TI - Pulse wave analysis and pulse wave velocity: a critical review of their strengths and weaknesses. AB - The study of the pulse using the technique of applanation tonometry is undergoing a resurgence with the development of new computerized equipment. We aim here to present a critical review of the uses, potential uses, strengths and weaknesses of the technique of applanation tonometry for the assessment of augmentation index and pulse wave velocity. We will review the technique of applanation tonometry, the physiological factors affecting pulse wave velocity and pulse wave analysis, the changes in pulse wave velocity and pulse wave analysis with pharmacological interventions, and the use of the technique of applanation tonometry as a prognostic tool. We conclude that, although the technique of applanation tonometry initially seems promising, several pertinent issues need to be addressed before it can be used reliably as a clinical or research tool. Importantly, use of the technique of applanation tonometry to derive the central waveform from non-invasively acquired peripheral data needs to be validated prospectively. PMID- 12640233 TI - Defining hypertension: never as simple as it seems. PMID- 12640234 TI - Obesity and hypertension in coronary heart disease: two independent PRIMErs? PMID- 12640235 TI - Determinants of heart rate and heart rate variability. PMID- 12640236 TI - White coat effect: semantics, assessment and pathophysiological implications. PMID- 12640237 TI - Arterial stiffness, gender and heart rate. PMID- 12640238 TI - Effects of caffeine on arterial function and haemodynamics: implications for cardiovascular risk. PMID- 12640239 TI - How to assess central arterial blood pressure? PMID- 12640240 TI - Second messengers in primary hypertension. PMID- 12640241 TI - Do COX-2 inhibitors really prevent hypertension and proteinuria? Another brick in the wall for the COX-2 inhibition and cardiovascular disease controversy. PMID- 12640242 TI - Subcutaneous resistive arteries and coronary conductance. PMID- 12640243 TI - Hypertension in the young: a therapeutic perspective? PMID- 12640244 TI - Assessing the prevalence of hypertension in populations: are we doing it right? AB - BACKGROUND: Although it is well recognized that the diagnosis of hypertension should be based on blood pressure (BP) measurements taken on several occasions, notably to account for a transient elevation of BP on the first readings, the prevalence of hypertension in populations has often relied on measurements at a single visit. OBJECTIVE: To identify an efficient strategy for assessing reliably the prevalence of hypertension in the population with regards to the number of BP readings required. DESIGN: Population-based survey of BP and follow-up information. SETTING AND PARTICIPANTS: All residents aged 25-64 years in an area of Dar es Salaam (Tanzania). MAIN OUTCOME MEASURES: Three BP readings at four successive visits in all participants with high BP (n = 653) and in 662 participants without high BP, measured with an automated BP device.RESULTS BP decreased substantially from the first to third readings at each of the four visits. BP decreased substantially between the first two visits but only a little between the next visits. Consequently, the prevalence of high BP based on the third reading--or the average of the second and third readings--at the second visit was not largely different compared to estimates based on readings at the fourth visit. BP decreased similarly when the first three visits were separated by 3-day or 14-day intervals. CONCLUSIONS: Taking triplicate readings on two visits, possibly separated by just a few days, could be a minimal strategy for assessing adequately the mean BP and the prevalence of hypertension at the population level. A sound strategy is important for assessing reliably the burden of hypertension in populations. PMID- 12640245 TI - Body mass index, hypertension and 5-year coronary heart disease incidence in middle aged men: the PRIME study. AB - OBJECTIVE: The aim of the present study was to assess the joint contribution of hypertension and body mass index to coronary heart disease risk. DESIGN Prospective study on men aged 50-59 years free of coronary heart disease at entry recruited in three regions of France (n = 7359) and in Northern Ireland (n = 2399). PARTICIPANTS: The recruitment frame was based on industry and various employment groups, on health screening centers and general practice. MAIN OUTCOME: Incident cases of effort angina, unstable angina, myocardial infarction and coronary death were recorded over a 5-year follow-up. RESULTS: Compared with the reference group [body mass index (BMI) 25 kg/m2], the relative risk of coronary event was higher in the second (25 < or =BMI 27.6) and third BMI tertiles: 1.27 (95% confidence interval 0.94-1.70) and 1.14 (0.84-1.56) after adjustment for confounders and covariates, including diabetes, hypertension and lipoprotein levels. Further analyses revealed a significant interaction between hypertension and BMI on coronary disease risk (P <0.05), suggesting that hypertension modifies coronary heart disease (CHD) risk attributable to BMI. Among hypertensive men, the relative risk of coronary heart disease was 1.34 (0.85-2.11) and 1.61 (1.04-2.50) in the second and third BMI tertiles, respectively. In normotensive men, BMI was not associated with CHD risk; relative risk 1.25 (0.85-1.85) and 0.66 (0.40-1.09) in the second and third BMI, respectively. CONCLUSION: These results indicate that hypertension and overweight jointly increase coronary heart disease risk. PMID- 12640246 TI - Host and environmental determinants of heart rate and heart rate variability in four European populations. AB - OBJECTIVE: In a population-based sample of nuclear families recruited in the framework of the European Project on Genes in Hypertension (EPOGH), we investigated the association between heart rate (HR) and its variability (HRV), and gender, age, posture, breathing frequency, body mass index, systolic blood pressure, family history of hypertension and various lifestyle factors, such as smoking, alcohol and coffee consumption and physical activity. METHODS: RR interval and respiration were registered in the supine and standing positions (15 min each) in 1208 subjects in Bucharest (Romania, n= 267), Cracow (Poland, n= 323), Mirano (Italy, n= 323) and Novosibirsk (Russian Federation, n= 295). After exclusion of 199 participants on antihypertensive treatment and/or patients with diabetes mellitus (n= 40) or myocardial infarction (n= 4), 993 subjects were eligible for analysis. We evaluated 858 participants with high-quality recordings. Using fast Fourier transform, we decomposed HRV into low-frequency (LF: 0.04-0.15 Hz) and high-frequency (HF: 0.15-0.40 Hz) components, which were expressed in normalized units. RESULTS: Mean values were 35.3 years for age, 24.3 kg/m for body mass index (BMI) and 121.0/77.2 mmHg for blood pressure. The group included 462 (53.8%) women. Across four centres, HR and HRV were similarly and independently associated with gender, age and postural position (P <0.001). In the supine position, HR was higher in women than men (67.2 versus 63.7 bpm). Men had higher normalized LF power than women (48.8 versus 41.5), but lower HF power (40.6 versus 47.4). The normalized HF power decreased with age (r = -0.43), whereas LF power increased (r = 0.32). On standing, HR increased (83.3 versus 65.6 bpm), normalized HF power declined (19.2 versus 44.3) and LF power increased (67.4 versus 44.9). The independent effects of respiration frequency, systolic blood pressure, family history of hypertension, body mass index and lifestyle factors on HRV differed between populations, and explained no more than 8% of the total variance. CONCLUSIONS: Across four European populations, gender, age and posture were consistent and independent correlates of HR and HRV. Lifestyle seems to have small but varying influences on HR and/or HRV, probably depending on the environmental and cultural background of the population under study. PMID- 12640247 TI - Influence of height, leg and trunk length on pulse pressure, systolic and diastolic blood pressure. AB - OBJECTIVE: To compare the effects of height, leg and trunk length on pulse pressure (PP), systolic (SBP) and diastolic blood pressure (DBP) in men and women. DESIGN: Prospective, population-based, birth cohort study. SETTING: England, Scotland and Wales. PARTICIPANTS: A total of 1472 men and 1563 women aged 53 years and followed since their birth in 1946. MAIN OUTCOME MEASURES: PP, SBP and DBP at age 53 years. RESULTS: PP increased linearly with decreasing height and leg length in men and women [shortest compared with the tallest height (leg length) group, men 54.6 versus 51.0 mmHg (55.8 versus 50.7 mmHg), women 52.9 versus 48.4 mmHg (53.3 versus 48.6 mmHg); P for trend < or = 0.001 in each case]. Adjustment for adult confounding factors and childhood social class or birth weight only slightly weakened these results (P for trend < or = 0.01 in each case). SBP, but not DBP, showed similar but weaker trends in men and women, except for the association with height in men, which lost statistical significance after adjustment. Trunk length was not associated with any outcome measure in men or women. CONCLUSIONS: Short height and leg length are associated with increased PP and SBP, but not DBP, in middle-aged men and women. PP is a potential mediator between short height and leg length and increased risk of coronary heart disease. Future studies of the association between shortness and coronary heart disease should consider the role of arterial hemodynamics. PMID- 12640248 TI - The white-coat effect is unrelated to the difference between clinic and daytime blood pressure and is associated with greater reactivity to public speaking. AB - OBJECTIVE: To compare the blood pressure (BP) response to doctor's visit with the BP reaction to a psycho-social challenge and with the difference between clinic and daytime BP (DeltaC-D). SUBJECTS: We studied 64 young stage-1 hypertensive subjects and 33 normotensive controls. MAIN OUTCOME MEASURES: Relationship between direct and surrogate measure of white-coat effect (WCE) and assessment of BP response to public speaking in subjects with normal or increased reaction to BP measurement. METHODS: The responses to BP measurement by a doctor and to public speaking were assessed with beat-to-beat Finapres recording. DeltaC-D was calculated on the basis of two BP monitorings and used as a surrogate measure of WCE. RESULTS: BP and heart rate changes elicited by the visit were unrelated to DeltaC-D and were correlated to the changes caused by the speech test [P <0.001 for systolic BP (SBP), P = 0.01 for diastolic BP (DBP), and P <0.001 for heart rate]. Hypertensive subjects with SBP response to doctor's visit above the median (hyper-reactive) showed increased reactivity also to public speaking (61 +/- 15 mmHg), while those with BP response below the median (normo-reactive) had a response to the psycho-social challenge (40 +/- 21 mmHg, 0.001 versus hyper reactive) similar to that of the normotensive controls (38 +/- 17 mmHg). Epinephrine urinary output was greater in the hyper-reactive than the normo reactive subjects (23 versus 12 microg/24 h, = 0.01). The SBP response to public speaking was greater in the hypertensive subjects with higher systolic daytime BP than in those with lower daytime BP (55.3 +/- 20.9 versus 45.1 +/- 20.6 mmHg, = 0.046). CONCLUSIONS: Subjects with increased WCE have an exaggerated response also to psycho-social stimuli. Average daytime BP, which incorporates the BP reactions to many psycho-social triggers can, thus, not be taken as the basal BP of an individual. This helps explain why DeltaC-D does not reflect the true WCE. PMID- 12640249 TI - Influence of sex on the relation between heart rate and aortic stiffness. AB - BACKGROUND: Heart rate and aortic pulse wave velocity (PWV) are both cardiovascular risk factors. The aim of this study was to evaluate the influence of sex on the heart rate-PWV relationship in two populations of normotensive and hypertensive men and women. PATIENTS AND METHODS: In a first study, steady-state data describing the heart rate-PWV relationship were determined in 558 normotensive men, 308 normotensive women, 323 hypertensive men and 93 hypertensive women. In a second study, the changes in blood pressure and PWV under atrial pacing at 60, 80 and 100 beats/min were investigated in 15 men and 15 women who were either normotensive or hypertensive. RESULTS: In women in study 1, 26.8% of PWV variance in normotensive patients and 15.4% of PWV variance in hypertensive patients was explained by a multiple regression including only age and systolic blood pressure as covariates. In men in study 1, 27% of PWV variance in normotensive patients and 28.5% of PWV variance in hypertensive patients was explained by a multiple regression including age, systolic blood pressure, weight, and also heart rate, as covariates. In study 2, atrial pacing in the overall population tended to decrease mean blood pressure (P = 0.05) and increase pulse pressure (P = 0.003), with no substantial change in PWV. However, when heart rate increased, PWV tended to decrease in women and to increase in men (interaction: P = 0.07).CONCLUSION Accelerated heart rate influences PWV in both normotensive and hypertensive men and, through this mechanism, could influence cardiovascular risk. However, heart rate does not influence PWV in women. PMID- 12640250 TI - Effect of caffeine on aortic elastic properties and wave reflection. AB - OBJECTIVE: Caffeine is the most widely used pharmacologically active substance. Aortic elastic properties and arterial wave reflection are important factors for the efficient performance of the cardiovascular system, as well as prognosticators of cardiovascular risk. We investigated the effect of caffeine on aortic elastic properties and wave reflection. DESIGN: We studied the effect of caffeine (250 mg) in 20 healthy subjects according to a randomized, placebo controlled, double-blind, cross-over design. METHODS: Aortic stiffness was evaluated with carotid-femoral pulse wave velocity and wave reflection with augmentation index of the aortic pressure waveform. RESULTS: Pulse wave velocity increased (by 0.51 m/s, 0.001) denoting an increase in aortic stiffness. Augmentation index and augmented pressure increased (by 6.8%, and by 4.4 mmHg, respectively, P <0.001 for both) denoting increased wave reflections. Concurrently, both radial and aortic systolic, diastolic and pulse pressure increased significantly. However, this increase was 20 and 9% larger for aortic systolic pressure (at 30 and 60 min, P <0.001 and P <0.05, respectively) and 56% larger for aortic pulse pressure (at 30 min, P <0.001) compared with the corresponding upper limb values. This indicates that peripheral pressure measurements are not an accurate guide for the pressor effect of caffeine in central arteries. CONCLUSIONS: Caffeine affects unfavorably aortic stiffness and enhances wave reflections. This finding has implications for the impact of caffeine consumption on cardiovascular risk. PMID- 12640251 TI - Peripheral blood pressure measurement is as good as applanation tonometry at predicting ascending aortic blood pressure. AB - BACKGROUND: The Sphygmocor system purports to be able to assess ascending aortic blood pressure using a transfer function. It has been shown to be accurate when data obtained invasively are used, but has not been tested prospectively using data obtained non-invasively. OBJECTIVE: To investigate the accuracy of this equipment when measurements are obtained non-invasively, as would normally be the case in the clinic setting. DESIGN AND METHODS: The study was observational. Ascending aortic pressure measurements were taken simultaneously with radial artery pressure wave recordings for estimation of ascending aortic blood pressure, in 28 patients undergoing diagnostic cardiac catheterization. RESULTS: The transfer function in the Sphygmocor system significantly underestimated invasively measured systolic blood pressure [mean -7.23 +/- 10.07 mmHg; 95% confidence interval (CI) -3.3 to -11.14 mmHg, P = 0.001] and significantly overestimated invasively measured diastolic blood pressure (mean 12.20 +/- 7.14 mmHg; 95% CI 9.43 to 14.97 mmHg, P <0.001). Oscillometrically measured brachial systolic blood pressure was not significantly different from that measured invasively in the ascending aorta (mean 3.36 +/- 10.47 mmHg; 95% CI -0.69 to 7.43 mmHg, P = 0.1), but oscillometric measurement of brachial diastolic blood pressure gave a significant overestimation of that measured invasively (mean 11.70 +/- 7.18 mmHg; 95% CI 8.91 to 14.49 mmHg, P <0.001). CONCLUSIONS The transfer function in the Sphygmocor system is no better at estimating ascending aortic blood pressure than are standard peripheral blood pressure measurements. It may be necessary to derive a new transfer system that is based on data that are acquired entirely non-invasively. PMID- 12640252 TI - High-salt-induced increase in blood pressure: role of capsaicin-sensitive sensory nerves. AB - OBJECTIVE: To test the hypothesis that sensory afferents are significant functional components in preventing salt-induced increases in blood pressure. DESIGN AND METHODS: Neonatal Wistar rats were subcutaneously injected with 50 mg/kg capsaicin or vehicle on the first and second days of life. After weaning, male rats were divided into three groups and treated for 4 weeks with: control plus normal (0.5%, CON-NS) or high (4%, CON-HS) sodium diet, and capsaicin pretreatment plus HS diet (CAP-HS). Mean arterial pressure (MAP) and its response to bolus injection of calcitonin gene-related peptide (CGRP) and its antagonist, CGRP(8-37), were measured by carotid arterial catheterization. Radioimmunoassay was used to measure CGRP levels in plasma and dorsal root ganglia (DRG). Expression of components of the CGRP receptor, calcitonin receptor-like receptor (CRLR) and receptor activity modifying protein 1 (RAMP1), was determined by the use of Western blot analysis. RESULTS: Baseline MAP was increased in CAP-HS compared with CON-HS and CON-NS rats, and it was higher in CON-HS than in CON-NS rats. MAP response to exogenous CGRP was enhanced in CAP-HS and CON-HS than in CON-NS rats, but MAP response to CGRP(8-37) was increased only in CON-HS rats. Plasma CGRP levels were not different among three groups, but CGRP content in DRG was decreased in CAP-HS compared with CON-HS and CON-NS rats. CRLR expression in mesenteric resistance arteries was upregulated in CAP-HS and CON-HS compared with CON-NS rats, but RAMP1 content was not different among these groups. CONCLUSION: Chronic high-salt intake upregulates expression of mesenteric CGRP receptors without altering CGRP levels in plasma and DRG. Increased expression of mesenteric CGRP receptors may play a counter regulatory role in attenuating salt induced increases in blood pressure. PMID- 12640253 TI - Phospholipase C activity is enhanced in skin fibroblasts obtained from patients with essential hypertension. AB - OBJECTIVES: In human hypertension, the response of phospholipase C (PLC) to stimuli is enhanced in signal transduction where receptors are coupled to pertussis toxin-sensitive G protein. We investigated PLC activity and its role in human hypertension. METHODS AND RESULTS: Skin fibroblasts were cultured from 15 normotensives subjects (53 +/- 4 years, four men and 11 women) and 19 essential hypertension (EH) patients (58 +/- 2 years, nine men and 10 women). Plasma membrane PLC activity, assessed by conversion of the tritiated exogenous phosphatidylinositol-4,5-bisphosphate to inositol trisphosphate, was greater in EH patients than in normotensive subjects (1.4 +/- 0.2 versus 0.7 +/- 0.1 pmol/mg protein/min, P <0.05). There was a positive correlation between PLC activity and mean blood pressure measured at admission and 7 days after admission (r = 0.47 and 0.37 respectively, both P <0.05). The value of the Michaelis constant was lower in EH patients than in normotensive subjects (32.1 +/- 5.6 versus 58.3 +/- 10.0 micromol/l, P <0.05), despite the fact that maximal velocity of the reaction was no different. Western blot analysis against PLC beta2 and beta3, gamma, delta1, and G protein gamma2 and gamma5 revealed that most PLC and G protein isoforms detected were delta1 of PLC and gamma2 of G protein, and no difference was detected in their amount between two groups. CONCLUSIONS: We conclude that enhanced PLC delta1 activity may be involved in the pathogenesis of human hypertension. PMID- 12640254 TI - Baroreflex and chemoreflex function after bilateral carotid body tumor resection. AB - OBJECTIVE: To investigate whether bilateral carotid body tumor resection invariably and chronically affects arterial baroreflex or peripheral chemoreflex function. METHODS: We studied eight consecutive patients (two men and six women; ages 48.1 +/- 11.8 years), a median time of 3.4 years (range 1.3-20.6 years) after bilateral carotid body tumor resection, and 12 healthy control individuals (eight men and four women; ages 53.7 +/- 10.1 years). Baroreflex sensitivity (phenylephrine), blood pressure and its variability (24 h Spacelabs and 5 h Portapres recordings), responses to standard cardiovascular reflex tests and the ventilatory responses to normocapnic and hypercapnic hypoxia were assessed. RESULTS: Baroreflex sensitivity was lower in patients (6.4 +/- 7.2 ms/mmHg) than in controls (14.7 +/- 6.6 ms/mmHg; P +/- 0.011). Mean office blood pressure and heart rate were normal in patients (123.3 +/- 11.9/79.0 +/- 7.3 mmHg and 67.5 +/- 9.4 beats/min, respectively) and controls (117.8 +/- 10.6/74.0 +/- 6.8 mmHg and 61.1 +/- 9.2 beats/min, respectively). Blood pressure variability was increased during ambulatory measurements. Three patients exhibited orthostatic hypotension. The Valsalva ratio, an index of baroreflex-mediated cardiovagal innervation, was lower in patients (1.4 +/- 0.2) than in controls (1.8 +/- 0.5; P +/- 0.008). The normocapnic ventilatory response to hypoxia was absent in all patients, whereas a small residual response to hypoxia was observed under hypercapnic conditions in two patients. CONCLUSIONS: Bilateral carotid body tumor resection results in heterogeneous expression of arterial baroreflex dysfunction, whereas the normocapnic hypoxic drive is invariably abolished as a result of peripheral chemoreflex failure. PMID- 12640255 TI - Inhibition of COX-2 prevents hypertension and proteinuria associated with a decrease of 8-iso-PGF2alpha formation in L-NAME-treated rats. AB - BACKGROUND: The inhibition of nitric oxide (NO) exerts injurious effects on the cardiovascular system by several mechanisms, such as the activation of the renin angiotensin system, oxidative stress, and inflammatory cytokines. We examined whether COX-2, an inducible isoform of cyclooxygenase, is associated with the pathogenesis observed in N(omega)-nitro-L-arginine methyl ester (L-NAME)-induced hypertensive rats. METHODS: Three groups of 8-week-old male Sprague-Dawley rats were studied (n = 6 in each group): group 1, untreated controls; group 2, treated with L-NAME (1 g/l for 3 weeks, p.o.); and group 3, L-NAME co-treated with COX-2 inhibitor NS-398 (5 mg/kg per day, i.p.). The L-NAME-induced expression of COX-2 mRNA and protein was semi-quantified in the kidneys and the thoracic aorta. Urinary excretion of the prostaglandin 6-keto PGF(1alpha), thromboxane B2 (TXB2), and prostaglandin E2 (PGE2) was measured in the three groups. Moreover, urinary excretion of 8-iso-PGF(2alpha), a potent vasoconstricting arachidonic acid metabolite acting through thromboxane A (TXA) receptor activation, proposed recently as a marker of oxidative stress, was also measured. RESULTS: L-NAME induced significant increases in systolic blood pressure (P< 0.01), urinary protein (P< 0.05), and renal excretion of 8-iso-PGF(2alpha)(P< 0.01), compared with the control. In L-NAME-treated rats, the levels of COX-2 mRNA and protein were more than 50% higher in the kidneys (P< 0.05), and six-fold higher in the thoracic aorta (P< 0.01) than in control rats. NS-398 significantly ameliorated an increase in systolic blood pressure (P< 0.01) and urinary protein (P< 0.05) induced by L-NAME. CONCLUSIONS: These data indicate that an increase in COX-2 expression might have a hypertensive effect, partly associated with 8-iso PGF(2alpha) formation in l-NAME-treated rats. PMID- 12640256 TI - Cardiovascular protective role of a low-dose antihypertensive combination in obese Zucker rats. AB - OBJECTIVE: Obesity, non-insulin-dependent diabetes mellitus (NIDDM) and hypertension are leading causes associated with increased cardiovascular morbidity and mortality. In modern times, the combined first line antihypertensive therapy with at least two drugs with a different mechanism of action to achieve a better blood pressure control, is increasing in acceptance worldwide. The aim of the present study was to determine possible beneficial effects of the low-dose combination (LDC) of an angiotensin-converting enzyme (ACE) inhibitor, perindopril (PER), and the diuretic indapamide (IND), regarding myocardial and vessels protection in an animal model of hypertension, obesity and NIDDM, such as the obese Zucker rat (OZR), and control lean Zucker rats (LZR). DESIGN: Ten-week-old male OZR (fa/fa) and LZR (Fa/fa) were used in this study. OZR group (G1, n=8), OZR with LDC group (G2, n=8); LZR group (G3, n=8) and LZR with LDC group (G4, n=8). During 6 months, G2 and G4 received a daily dose of 1 mg/kg combination of 0.76 mg/kg PER + 0.24 mg/kg IND, (ratio of doses 0.32), by gavage, and G1 and G3 received an equal volume of vehicle throughout the experiment. In order to evaluate cardiac dimensions and left ventricular mass (LVM) transthoracic echocardiograms were performed, at baseline and at the end of the experiment. Urine and blood samples for biochemical determination were obtained. After 6 months of treatment all rats were sacrificed, hearts were harvested for light microscopy (LM), high-resolution light microscopy (HRLM), immunohistochemistry including monoclonal antibodies against transforming growth factor beta (TGFbeta1) and anti-collagen type I (COL I) and type III (COL III) and electron microscopy (EM) studies. RESULTS: At the end of the study OZR treated with LDC presented: (1) lower blood pressure (128.9 +/- 4 versus 150.3 +/ 3.6 mmHg, P< 0.05); (2) smaller cardiac dimensions (P< 0.01); (3) lower LVM/100 g body weight (0.17 +/- 0.02 versus 0.30 +/- 0.05, P< 0.01); (4) higher fractional shortening (34.5 +/- 3.2 versus 23.3 +/- 5.9%, P< 0.01) than OZR untreated. Moreover, OZR that received LDC showed higher: (1) myocyte density (48 +/- 1.5 versus 20 +/- 2.5 myocytes/area, P< 0.01); (2) capillary density (30.5 +/ 3.1 versus 9.5 +/- 1.6 capillaries/area, P<0.01); (3) myofilament thickness (12.05 +/- 0.27 versus 9.83 +/- 0.39 nm, P<0.01); and lower amounts of: (1) TGFbeta1 in myocytes (P< 0.01), interstitium (P< 0.01) and vessel wall (P< 0.05); (2) COL I and COL III (P< 0.01), and COL I /COL III ratio (P< 0.01), compared with untreated OZR. Finally, OZR-treated with LDC showed not only unsubstantial modification in carbohydrate and lipid metabolism when compared with untreated OZR, but also an improvement in insulin/glucose ratio (P< 0.05). CONCLUSION: These results suggest that LDC of PER + IND can control cardiovascular damage in OZR providing an additional help in the metabolic scenario likewise. PMID- 12640257 TI - B2 bradykinin receptor (B2BKR) polymorphism and change in left ventricular mass in response to antihypertensive treatment: results from the Swedish Irbesartan Left Ventricular Hypertrophy Investigation versus Atenolol (SILVHIA) trial. AB - OBJECTIVE: Hypertension is associated with a number of adverse morphologic and functional changes in the cardiovascular system, including left ventricular (LV) hypertrophy. Studies have demonstrated that bradykinin, through the B2 bradykinin receptor (B2BKR), mediates important cardiovascular effects that may protect against LV hypertrophy. Recently, a +9/-9 exon 1 polymorphism of the B2BKR was shown to be strongly associated with LV growth response among normotensive males undergoing physical training. We aimed to clarify whether the processes found in exercise-induced LV growth in normotensive people also occur in pathological LV hypertrophy. DESIGN AND METHODS: We determined the B2BKR genotype of 90 patients with essential hypertension and echocardiographically diagnosed LV hypertrophy, included in a double-blind study to receive treatment for 48 weeks with either the angiotensin II type 1 (AT1) receptor antagonist irbesartan or the beta1 adrenoceptor antagonist atenolol. RESULTS: B2BKR +9/+9 genotypes responded poorly in LV mass regression, independent of blood pressure reduction or treatment, as compared to the other genotypes (adjusted mean change in LV mass index = -10.0 +/ 4.6 versus -21.6 +/- 2.2 g/m2, P = 0.03). CONCLUSIONS: Our results suggest an impact of the B2BKR polymorphism on LV mass regression during antihypertensive treatment. PMID- 12640258 TI - Relationships between coronary flow vasodilator capacity and small artery remodelling in hypertensive patients. AB - BACKGROUND: Arterial hypertension is frequently associated with the presence of structural alterations in small arteries. Moreover, a reduced coronary flow reserve and vasodilator capacity has been observed in essential hypertensive patients, possibly due, at least in part, to microangiopathy of small coronary vessels. The aim of the present study was to evaluate a possible relationship between subcutaneous small artery structure and coronary flow reserve or vasodilator capacity in patients with essential hypertension. METHODS AND RESULTS: A total of 20 patients with mild to moderate essential hypertension were included in the study, and underwent a biopsy of the subcutaneous fat from the gluteal region. Small arteries were dissected and mounted on a micromyograph. The media thickness, the normalized internal diameter and the media:lumen ratio (M/L) were then calculated. In addition, a transesophageal Doppler echocardiographic study, which allows the measurement of coronary flow velocity before and during maximal pharmacological vasodilatation, was performed. Coronary flow reserve (CFR) was measured as the ratio of coronary flow velocity assessed during adenosine infusion and that measured in basal conditions. From blood pressure and coronary flow velocity during adenosine infusion, minimum coronary resistance was calculated. CFR as well as minimum coronary resistance were significantly correlated to both M/L and to normalized internal diameter of subcutaneous small arteries. CONCLUSIONS: Our results are consistent with the hypothesis of a generalized remodelling of small arteries in the body, including the coronary circulation; this remodelling may play an important role in the reduction of coronary vasodilator capacity in patients with mild to moderate essential hypertension. PMID- 12640259 TI - How good is blood pressure control among treated hypertensive children and adolescents? AB - BACKGROUND: Since good control of arterial hypertension is of paramount importance, the present study was carried out to evaluate blood pressure control in pediatric patients with hypertension receiving regular medical care. STUDY DESIGN: The charts of 80 hypertensive children receiving medical care were reviewed. Their antihypertensive medication had been stable during three or more separate clinic visits and during 3 or more months. Patients with office hypertension were excluded. RESULTS: Blood pressure values higher than the corresponding 95th centiles were noted in 20 of the 80 patients. Hypertension was systolic in seven, diastolic in four and both systolic and diastolic in nine patients. The number of prescribed antihypertensive drugs and the number of doses/day of prescribed antihypertensive drugs was similar in patients with good and in those with poor blood pressure control. Plasma creatinine was higher in patients with poor than in those with good blood pressure control. CONCLUSIONS: The present survey indicates that the goal of antihypertensive medication is not achieved in a noticeable number of pediatric patients with treated hypertension. PMID- 12640260 TI - Mental stress and hypertension, an evolutionary framework: some historical perspectives of the 1960 World Health Organization Prague Hypertension Meeting. AB - Emotional stress acutely and repetitively causing blood pressure increase or aggravating existing hypertension is usually not reflected by norepinephrine and epinephrine increase but by a sudden rise of dopamine, the third "defensive" catecholamine coping with the damaging neuropsychological and cardiovascular actions of the first two. This double-edged sympathetic response to emotional stress evolves during human lifespan and long-term evolution of hypertension. In the course of philogenesis it carries a potential mismatch between the normal physiology of the human dopaminergic system and current environmental (emotional particularly) conditions in industrialized countries. This offers a rational support to a mental stress-cardiovascular diseases relationship proposed 40 years ago in a WHO report which followed a memorable 1960 Prague Hypertension Meeting. PMID- 12640261 TI - The vascular amplifier and persisting resisters. PMID- 12640262 TI - Estimation of 24-h urinary sodium excretion using lean body mass and overnight urine collected by a pipe-sampling method. PMID- 12640269 TI - Science versus art of medicine. PMID- 12640270 TI - Pulmonary surfactant for neonatal respiratory disorders. AB - Surfactant therapy has revolutionized neonatal care and is used routinely for preterm infants with respiratory distress syndrome. Recent investigation has further elucidated the function of surfactant-associated proteins and their contribution toward surfactant and lung immune defense functions. As the field of neonatology moves away from intubation and mechanical ventilation of preterm infants at birth toward more aggressive use of nasal continuous positive airway pressure, the optimal timing of exogenous surfactant therapy remains unclear. Evidence suggests that preterm neonates with bronchopulmonary dysplasia and prolonged mechanical ventilation also experience surfactant dysfunction; however, exogenous surfactant therapy beyond the first week of life has not been well studied. Surfactant replacement therapy has been studied for use in other respiratory disorders, including meconium aspiration syndrome and pneumonia. Commercial surfactant preparations currently available are not optimal, given the variability of surfactant protein content and their susceptibility to inhibition. Further progress in the treatment of neonatal respiratory disorders may include the development of "designer" surfactant preparations. PMID- 12640272 TI - The controversies surrounding oxygen therapy in neonatal intensive care units. AB - Despite the knowledge that excess amounts of oxygen in the blood (hyperoxia) can be damaging to preterm infants, there is a wide variation in approaches to oxygen therapy within neonatal intensive care units. This is predominantly determined by institutional or individual practices or preferences and might stem from a lack of understanding of the relative merits and demerits of the different techniques of oxygen monitoring in extremely preterm babies who are different from more mature babies. This article provides the physiological rationale and evidence from recent clinical studies suggesting that keeping the oxygen therapy to an "acceptable" minimum in premature babies does not do any harm and may be even advantageous. PMID- 12640271 TI - Immunotherapy in the prophylaxis and treatment of neonatal sepsis. AB - Neonatal sepsis is a significant cause of morbidity and mortality in the neonatal intensive care unit. The epidemiology of neonatal infections is complex; however, they are in large part secondary to developmentally immature host defense mechanisms. These immunodeficiencies, which are exaggerated in premature and sick neonates, include quantitative and qualitative deficits in phagocytes, complement components, cytokines, and immunoglobulins. Therapies that modulate or augment host defenses may attenuate the virulence of neonatal infections. In this paper, we have reviewed immunotherapies that modulate the immune system of the neonate, including: intravenous immunoglobulins, myeloid hematopoietic growth factors, and granulocyte transfusions. Future studies should focus on investigating other abnormalities of neonatal host defense and/or combined immunotherapy approaches in an attempt to circumvent the immaturity of host defense and potentially reduce both the incidence and severity of neonatal sepsis. PMID- 12640273 TI - Fetal origins of cardiovascular disease. AB - Several epidemiologic studies have shown that intrauterine growth retardation is a risk factor for the development of cardiovascular disease in later life. In this review, we discuss these epidemiologic studies and animal models that have been developed to investigate the pathophysiology of this phenomenon. We discuss data suggesting that intrauterine growth retardation leads to fetal exposure to maternal glucocorticoids. In addition, we present other data showing that fetal exposure of glucocorticoids during specific times of fetal development results in focal and segmental glomerulosclerosis, a reduced number of nephrons, hypertension, and diabetes. These studies suggest that at critical times during fetal development fetal injury programs the development of cardiovascular disease and diabetes in later life. PMID- 12640274 TI - Focal segmental glomerulosclerosis: pathogenesis and treatment. PMID- 12640276 TI - New imaging applications in the evaluation of pediatric renal disease. AB - Continued improvements in imaging technology have changed many of the traditional diagnostic algorithms for evaluating pediatric renal disease. Newer imaging modalities offer more accurate, specific, and early diagnoses, but can be time consuming and costly. Less invasive modalities, such as ultrasound, computed tomography, and magnetic resonance imaging have widespread applications in pediatric practice. The risks of radiation exposure, contrast toxicity, and sedation or anesthesia versus the potential benefits of obtaining precise diagnostic information should always be considerations before electing any imaging procedure in children. PMID- 12640275 TI - Primary ureteral reflux: emerging insights from molecular and genetic studies. AB - The incidence of primary vesicoureteral reflux is about 1% to 2% of the general population and is as high as 50% in siblings as well as offspring of affected patients, suggesting autosomal dominant inheritance. The current diagnosis of vesicoureteral reflux involves voiding cystourethrograms, which are invasive and costly. Consequently, vesicoureteral reflux screening in siblings and offspring is not routinely practiced, because of the known high risk. Early detection of vesicoureteral reflux will be valuable for prevention of reflux nephropathy, because the incidence of reflux nephropathy can be reduced effectively by antibiotic prophylaxis. Furthermore, the presence of reflux nephropathy can only be accurately assessed currently by dimercapto-succinic acid nuclear scans, which are costly, time and labor intensive, and often require conscious sedation by a pediatric anesthesiology team. As a result, the clinical assessment of reflux nephropathy is also not routinely practiced. There is a pressing need to develop less invasive and less costly tests for the early diagnosis of primary vesicoureteric reflux and reflux nephropathy. Recent molecular and genetic studies have greatly increased our understanding of vesicoureteral reflux and provide a promise of novel non-invasive tests. Targeted disruption of angiotensin type II receptor and uroplakin III genes result in the phenotype of primary vesicoureteral reflux. There are characteristic patterns of message and protein changes in the knockout animals, providing the basis for detection of genetic mutations leading to vesicoureteral reflux in humans by studying differential gene expression by functional genomics methodology. The urothelium is also known to secrete proteins into the urine. Preliminary studies showed unique fingerprints in urinary protein patterns in children with primary VUR, providing the basis for developing novel noninvasive molecular diagnostic tests of vesicoureteral reflux by proteomics methodology. PMID- 12640278 TI - Amiodarone-an "old" drug with new recommendations. AB - Amiodarone has gained recognition as an antiarrhythmic medication after recent publication of the newly revised American Heart Association guidelines for pediatric resuscitation. Although support for the widespread use of amiodarone in adults has been supported by research, the few pediatric studies demonstrate limited efficacy and highlight the need for additional data. Because of the nature of the need for this type of resuscitation medication, controlled prospective studies will be difficult to obtain if not morally contraindicated. This article reviews the properties of amiodarone and the pertinent pediatric studies to provide healthcare providers supplemental information regarding amiodarone when choosing antiarrhythmics for acute resuscitation. Individual providers need to discern whether the pediatric data available supports widespread acceptance into current treatment regimens. PMID- 12640279 TI - Newer pharmacologic agents for procedural sedation of children in the emergency department-etomidate and propofol. AB - Procedural sedation for pediatric patients having painful or anxiety-producing procedures is a necessary but often a daunting task for emergency medicine providers. This article focuses on the two agents that have most recently been described for use in this population-etomidate and propofol. Etomidate is a nonbarbiturate sedative hypnotic agent with no analgesic properties. Its rapid onset of action, short duration of action, and minimal hemodynamic effects make it an attractive agent for use in procedural sedation. Similar to previous adult studies, recent studies have shown that etomidate is both safe and effective in this pediatric population. Propofol is also a sedative hypnotic agent with rapid onset and short duration of action. Typically, it is administered as a bolus injection followed by an infusion. It has long been used for surgical procedures as well as in the intensive care unit setting, but little literature has supported its use in the pediatric emergency department. Recent studies appear to support propofol's use in this setting; however, a significant rate of side effects, including hypoxia, apnea, and decreased blood pressure, may limit its use. PMID- 12640280 TI - Inhalant abuse in the pediatric population: a persistent challenge. AB - Inhalants remain one of the most commonly abused substances by adolescents in the United States today. Recent epidemiologic studies have demonstrated a decrease in the prevalence of inhalant abuse, but overall rates remain high. Inhalants have the potential to result in serious organ system dysfunction or even sudden death. This article reviews the most recent epidemiologic data on inhalant abuse, summarizes the types of substances used and their mechanism of actions, and discusses clinical toxicity and medical management. PMID- 12640281 TI - The resurgence and abuse of heroin by children in the United States. AB - Heroin, one of the most addictive and "hardest" drugs of abuse, carries significant morbidity and mortality. Although its use is usually associated with the adult population in the United States, the last decade has witnessed a decrease in the median age of heroin users. An increase in the availability of inexpensive and pure heroin that could be snorted rather than injected made the drug accessible to adolescents and reduced the fear associated with the transmission of the human immunodeficiency virus (HIV). Because of the atypical demographics and the alternative drug use patterns, this young population of heroin users is not easily identified by parents or by healthcare providers. Lack of social support or access to healthcare prevents young heroin users from participation in detoxification programs, suggesting that changes may be needed in our healthcare and social systems to properly target and provide care to the youngest heroin abusers. PMID- 12640282 TI - Updates on attention deficit hyperactivity disorder, child abuse and neglect, and sudden infant death syndrome. PMID- 12640283 TI - A 24-day old with episodic lethargy, hypotonia, and apnea: the eyes have it. PMID- 12640284 TI - Long-term outcomes in patients with sickle cell disease and frequent vaso occlusive crises. AB - BACKGROUND: The frequency of vaso-occlusive crises correlates with mortality in patients with sickle cell disease (SCD). We examined the degree to which a high number of hospitalization days for these events affected survival. PATIENTS AND METHODS: We reviewed data for 58 adult patients with SCD (mean age, 29.9 +/- 7.3 years) treated at our hospital between 1986 and 1994 who had at least 100 hospitalization days during any of these years. Their mean follow up period was 6.65 years (median, 6.84 years; range, 0.15-14.51 years). RESULTS: Thirty-five patients (60.3%) died during follow-up. In multivariate analysis that included age, gender, and numbers of transfusions and hospitalization days, only age was significantly associated with mortality. The National Cooperative Study of Sickle Cell Disease (CSSCD) estimates the 10-year mortality at 15% for all 20-year old SS patients and also for all 30-year-old women. For 30-year-old men, the CSSCD estimated a 10-year mortality of about 28%. Thus, the 60.3% mortality of our patients after a median follow-up of only 6.84 years was substantially higher. Fifty-one patients were still alive after 1992, when hydroxyurea became available for SCD. The median survival of 15 hydroxyurea-treated patients was 7.3 years, compared with 4.3 years in 36 patients who did not take the drug. CONCLUSIONS: Mortality for patients with SCD with a high number of hospitalization days was much higher than that expected for patients with SCD in general. There was a (nonsignificant) trend for longer survival in these severely ill patients if they took hydroxyurea. PMID- 12640285 TI - Glomerular and tubular dysfunction in children with congenital cyanotic heart disease: effect of palliative surgery. AB - BACKGROUND: Nephropathy has long been recognized as a potential complication of congenital cyanotic heart disease (CCHD). The present study was undertaken to investigate some aspects of glomerular function by measuring urinary total protein, microalbumin, and tubular function by assessing urinary alpha-1 microglobulin. The structural integrity of the renal proximal tubules was also studied by measuring urinary activities of the brush-border enzyme leucine aminopeptidase and the lysosomal enzyme -acetyl-beta-d-glucosaminidase. The levels of hematocrit (Hct) and oxygen saturation were also investigated as predisposing factors for renal impairment in CCHD. METHODS: These investigations were done by recruiting 86 children who were grouped as follows: the control group (G1 ) consisted of 14 children (aged 4-12 years); the other 72 children with CCHD were divided according to age (ie, duration of cyanosis) into 4 equal groups, each containing 18 patients: G2 (age <1 year), G3 (age > or = 1 year and <5 years), G4 (age > or = 5 years and <10 years), and G5 (age > or = 10 years). In addition, 10 of the 72 patients underwent a palliative surgery and were included as G6 (regardless of age: 2 from G3, 4 from G4, and 4 from G5 ) to study the effect of the palliative surgery on the above-mentioned parameters. RESULTS: Results of the present work showed that with increasing duration of cyanosis (ie, on going from G2 to G5 ) among the studied children with CCHD, there was a significant elevation in the urinary excretion of the investigated functional and structural parameters of the glomeruli and proximal tubules compared with the control children. The data also showed a significant increase in Hct, whereas oxygen saturation was significantly decreased. Results of G6 after the palliative surgery demonstrated a significant decrease in the urinary excretion of the investigated parameters of the kidney, with a significant decrease in Hct and increase in oxygen saturation levels, compared with the results of the patients of this group before the palliative surgery. CONCLUSIONS: These results suggest impairment of both glomerular and tubular functions as well as structure of the proximal tubules among children with CCHD and that the palliative surgery has significantly improved this impairment. PMID- 12640286 TI - Isometric exercise increases the size of forearm veins in patients with chronic renal failure. AB - OBJECTIVES: Delay in maturation or failure of maturation of Cimino-Brescia fistulae contributes to the significant vascular access-related morbidity of chronic hemodialysis patients. Increased size and capacitance of native veins before the formation of vascular access has been considered an important variable in the success rate of native fistulae. We evaluated whether a formal exercise program might alter the size of native veins. METHODS: The effect of exercise on venous size was evaluated in 5 patients with severe chronic renal failure [glomerular filtration rate, 30.6 +/- 5.3 mL/min (mean +/- SD)]. Five male patients with a mean age of 57 +/- 9 years underwent a 6-week forearm exercise training program, involving nondominant arms, that included isometric hand-grip contractions to 25 to 35% of MVC lasting 40 to 120 seconds and repetitive squeezing of squash and racquet balls. Both the volume and intensity of exercise training was increased weekly based on strength measured by hand-grip dynamometer and on the patients' indicated level of comfort. Cephalic vessel size in both the nondominant (trained) and dominant (control) arms, with and without a tourniquet, were obtained using Doppler ultrasound before and after the 6-week exercise training program. RESULTS: The size of the cephalic vein of the exercised arm increased significantly ( < 0.05) compared with the control arm when measured in both the absence (048 +/- 0.016 versus 0.024 +/- 0.023 cm ) and the presence of a tourniquet (0.056 +/- 0.022 versus 028 +/- 0.027 cm ). CONCLUSIONS: These findings indicate that a simple, incremental resistance, exercise-training program can cause a significant increase in the size of the cephalic vein commonly used in the creation of an arteriovenous fistula. The increase in size and resultant probable increase in blood flow might accelerate the maturation of native arteriovenous fistulae, thereby lessening the morbidity associated with vascular access. PMID- 12640287 TI - The role of DNA testing in breast, ovarian, and colon cancers. AB - A proportion of cases of breast and colon cancers are caused by inherited mutations that confer a greatly increased susceptibility to malignancy. Certain clinical features may help distinguish patients with a genetic cause for their cancer from the larger number of patients with sporadic tumors. A thorough family medical history is also necessary to identify those at high risk for developing cancer. Many of the normal functions of these genes are understood, and mutation analysis for patients and their families is now available as a clinical service. Presymptomatic detection of mutations allows the patient to pursue preventive measures to reduce the probability of developing a malignancy. Evidence is now available that some prophylactic measures do reduce the incidence of cancer and reduce mortality in mutation carriers, and the standard of care is evolving rapidly. The essential elements necessary to provide accurate interpretations of molecular genetics test results to patients are described. PMID- 12640288 TI - Process of care and outcomes in peripheral arterial disease. AB - Peripheral arterial disease (PAD) is a prevalent illness that most commonly affects patients older than 60 years. As the population ages, the prevalence of PAD and its associated adverse outcomes will also increase. Adverse outcomes in PAD are either systemic (ie, cardiovascular events such as myocardial infarctions or strokes) or localized to the legs (ie, bypass surgery or amputation). Although much research has focused on adverse systemic outcomes in patients with PAD, less is known about those factors related to adverse limb events. The Centers for Disease Control and Prevention estimate that more than 100,000 amputations are performed in the United States each year. What remains to be determined is the association between how physicians manage patients with PAD (ie, process of care) and the development of adverse limb outcomes. Determining the association of the management of PAD with adverse limb outcomes will highlight those areas in which to focus to improve the quality of care for patients with this disease. Understanding the relationship between risk factors, process of care, and outcomes will be of importance to patients, clinicians, and policy makers. The purpose of this article is to review the burden of PAD and to discuss the association of process of care with adverse limb outcomes in patients with PAD. PMID- 12640289 TI - Extrahepatic manifestations of hepatitis C infection. AB - Hepatitis C virus (HCV) affects 1.8% of the American population, and approximately 38% of patients with HCV will manifest symptoms of at least 1 extrahepatic manifestation during the illness. Renal disease, neuropathy, lymphoma, and Sjogren syndrome with or without mixed cryoglobulinemia are all strongly associated with HCV infection. Porphyria cutanea tarda and diabetes have also been linked to HCV. Most extrahepatic manifestations of chronic HCV infection are immunological, and the chronic infection seems to be necessary for their development. The molecular study of the unique way in which the HCV virus interacts with the human immune system is beginning to provide plausible explanations of the pathogenic role of HCV in some of these syndromes, but many pathogenetic links remain completely obscure. PMID- 12640290 TI - Polycythemia vera responds to imatinib mesylate. AB - We report 2 patients with polycythemia vera who were demonstrated to be -negative and were unable to tolerate either hydroxyurea or interferon-alpha but who had excellent clinical responses to imatinib mesylate (STI-571). This effect is consistent with the inhibitory effect of imatinib mesylate on c-kit's tyrosine kinase activity as demonstrated by its effectiveness in patients with gastrointestinal stromal tumors. PMID- 12640291 TI - An unusual cause of hypokalemic paralysis: chronic licorice ingestion. AB - Long-term licorice ingestion is a well-known cause of secondary hypertension and hypokalemia. Nevertheless, its initial presentation with a very severe degree of hypokalemia and paralysis is exceedingly rare. We report an elderly Asian man who presented to the emergency department with marked muscle weakness that progressed to paralysis. His blood pressure was 160/96 mm Hg. The major biochemical abnormalities were hypokalemia (plasma K+ concentration, 1.8 mmol/L) and metabolic alkalosis (HCO - 3 , 36 mmol/L). His renal potassium excretion was higher (transtubular potassium gradient of 9). Plasma renin activity and aldosterone concentration were suppressed and cortisol concentration was normal. A detailed history revealed that he had ingested tea flavored with 100 g of natural licorice root containing 2.3% glycyrrhizic acid daily for 3 years. Note that renal potassium wasting and hypertension persisted for 2 weeks after discontinuing licorice consumption along with KCl supplement and spironolactone. Long-term licorice ingestion should be kept in mind as a cause of paralysis with an extreme degree of hypokalemia to avoid missing this recognizable and curable medical disorder. PMID- 12640292 TI - Metastatic melanoma presenting as cardiac mass and hemobilia. AB - We present a rare case of malignant melanoma of unknown primary origin presenting as cardiac and cholecystic masses. In this 72-year-old man, a cardiac lesion caused right ventricular outlet obstruction, and gallbladder lesions led to hemobilia. Right atrial and gallbladder metastases were resected. After 2 years of disease-free survival, the patient had recurrent skin lesions, which were treated with local resection and dacarbazine. The patient has survived for more than 5 years after initial diagnosis, with continued complete clinical remission. This case illustrates the importance of aggressive surgical resection and chemotherapy in the treatment of metastatic malignant melanoma. PMID- 12640293 TI - Prevention of ischemia-reperfusion-induced hepatic microcirculatory disruption by inhibiting stellate cell contraction using rock inhibitor. AB - BACKGROUND: We demonstrated that hepatic stellate cells (HSCs) isolated from rat livers exposed to warm ischemia are significantly contractile when compared with HSCs from intact rat livers. This suggests that ischemia-reperfusion (IR)-induced impairment of sinusoidal microcirculation results, at least in part, from contraction of HSCs. METHODS: Rho-associated coiled-coil forming protein serine/threonine kinase (ROCK) is one of the key regulators of HSCs motility. Therefore we investigated whether Y-27632, a p160ROCK-specific inhibitor, has beneficial effects on warm IR injury in an in vivo rat partial liver IR model and a rat orthotopic liver transplantation model. RESULTS: After reperfusion following 90 min of warm ischemia, livers in untreated control rats had persistent congestion and impaired mitochondrial respiration, as demonstrated by increasing deoxy-hemoglobin and reduced cytochrome oxidase contents in the hepatic tissues using in vivo near-infrared spectroscopy. Serum levels of transaminase and endothelin (ET)-1 in these rats were markedly increased 1 hr after reperfusion. In contrast, when Y-27632 (3-30 mg/kg) was administered orally, hepatic tissue contents of deoxy-hemoglobin and cytochrome oxidase rapidly normalized. In such animals, the elevation of serum transaminase levels, but not that of ET-1 levels, was significantly suppressed. This is consistent with in vitro data demonstrating that Y-27632 causes HSCs to undergo relaxation even in the presence of ET-1. Moreover, in a rat orthotopic liver transplantation model, Y-27632 pretreatment dramatically improved the survival of recipients with liver grafts subjected to 45 min of warm ischemia. CONCLUSIONS: Y-27632 attenuates IR-induced hepatic microcirculation disruption by inhibiting contraction of HSCs. PMID- 12640294 TI - Predominant expression of Th2 cytokines and interferon-gamma in xenogeneic cardiac grafts undergoing acute vascular rejection. AB - BACKGROUND: The Th1 response has been shown to play a role in acute allograft rejection, whereas the Th2 response has been implicated in the protection of allografts. Unlike allografts, the pattern of cytokines in response to solid organ xenografts has been the subject of limited studies. We investigated intragraft cytokine expression in a concordant cardiac xenograft model (rat-to mouse) to test if a particular cytokine profile predominates. METHODS: Intra abdominal cardiac transplantation was performed using C57BL/10 mice as recipients of PVG.R8 rat hearts. Syngeneic grafts (C57BL/10-to-C75BL/10) served as controls. Cardiac grafts harvested on various days posttransplantation were analyzed for histology and intragraft cytokine expression using reverse-transcriptase polymerase chain reaction. RESULTS: The grafts in this model were rejected with a mean survival time of 7+/-1 days and showed extensive evidence of acute vascular rejection, consisting of global distortion of myocardial architecture, fewer cellular infiltrates, interstitial hemorrhage with myocyte necrosis thrombosis, and vasculitis with neutrophils and lymphocytes infiltrating vessel walls. Cardiac xenografts predominantly expressed Th2 cytokines, interleukin (IL)-4, IL 10, and transforming growth factor-beta with various kinetics. IL-10 was detectable on day 1 and reached its peak level of expression on day 6 posttransplantation. IL-4 showed minimal and undetectable expression on days 1 and 3 and significant expression on day 6 posttransplantation. Transforming growth factor-beta was expressed moderately on all days examined. The expression of interferon (IFN)-gamma, a Th1 cytokine, was specific to xenografts and showed a gradual increase from days 3 to 6 posttransplantation. In marked contrast, IL-2 showed complete lack of expression. CONCLUSIONS: Our data demonstrate predominant expression of Th2 cytokines and IFN-gamma in cardiac xenografts undergoing acute vascular rejection. The Th2 cytokines may promote acute vascular rejection by regulating the humoral response, and IFN-gamma may delay, but not prevent, this response. PMID- 12640295 TI - Glycine ameliorates lung reperfusion injury after cold preservation in an ex vivo rat lung model. AB - BACKGROUND: The role of glycine has not been investigated in lung ischemia reperfusion injury after cold preservation. Furthermore, the role of apoptosis after reperfusion following cold preservation has not been fully understood. METHODS: Lewis rats were divided into three groups (n=6 each). In the GLY(-) and GLY(+) groups, isolated lungs were preserved for 15 hr at 4 degrees C after a pulmonary artery (PA) flush using our previously developed preservation solution (ET-K; extracellular-type trehalose containing Kyoto), with or without the addition of glycine (5 mM). In the Fresh group, isolated lungs were reperfused immediately after a PA flush with ET-K. They were reperfused for 60 min with an ex vivo perfusion model. Pulmonary function, oxidative stress, apoptosis, and tumor necrosis factor (TNF)-alpha expression were assessed after reperfusion. RESULTS: Shunt fraction and peak inspiratory pressure after reperfusion in the GLY(-) group were significantly higher than those in the GLY(+) and Fresh groups. Oxidative damage and apoptosis in the alveolar epithelial cells of the GLY(-) group, assessed by immunohistochemical staining and quantification of 8-hydroxy 2'-deoxyguanosine and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling method, were significantly higher than those of the GLY(+) and Fresh groups. There were correlations among shunt fraction, oxidative damage, and apoptosis. There was no expression of TNF-alpha messenger RNA in all groups evaluated by the reverse transcription-polymerase chain reaction. CONCLUSIONS: Glycine attenuates ischemia/reperfusion injury after cold preservation by reducing oxidative damage and suppressing apoptosis independent of TNF-alpha in this model. The suppression of apoptosis might ameliorate lung function after reperfusion. PMID- 12640296 TI - Islet transplantation in the discordant tilapia-to-mouse model: a novel application of alginate microencapsulation in the study of xenograft rejection. AB - BACKGROUND: Tilapia islet xenograft rejection is characterized by infiltration with macrophages (Mphis), eosinophils (Ephis), and T lymphocytes. The presence of these cells indicates they contribute to rejection; therefore, an attempt was made to assess their role through host immunomodulation. METHODS: Tilapia islet cells were transplanted under the kidney capsule of streptozotocin diabetic Balb/c mice, which were then treated with one of several immunomodulatory regimes targeting Mphis, Ephis, or T cells. Mphis were depleted using either silica or liposome-entrapped Cl2MDP. Ephi migration was blocked using monoclonal antibodies (mAbs) targeting interleukin (IL)-4 or IL-5. T-cell function was altered with mAbs targeting CD3, CD4, or CD8. Finally, T helper (Th)1 and Th2 activity was altered by depleting essential Th1 or Th2 cytokines with mAbs or by promoting a Th1 response with the injection of exogenous IL-12. The effects of antibody mediated immunomodulation on graft survival were initially screened by cotransplanting alginate-encapsulated, mAb-secreting hybridoma cells into the peritoneal cavity at the time of islet transplantation. Significant prolongation was then confirmed using purified antibodies injected at the time of islet transplantation. Rejected grafts were examined histologically, and immunohistochemistry was used to assess the cellular infiltrates for each of the treatment groups. RESULTS: Modulation of Mphis and Ephis alone did not significantly delay functional rejection of tilapia islet grafts (maximal mean graft survival time [mGST]=7.1+/-1.7 and 9.4+/-3.4, respectively) compared with untreated controls (mGST=8.2+/-1.0). Treatment of transplanted animals with antibodies against CD3 or CD4 significantly promoted graft survival (maximal mGST=16.3+/-5.8 and 34.0+/-11.6, respectively), whereas targeting CD8 and Th1 and Th2 cytokines showed no prolonging effect (maximal mGST=7.8+/-2.9 and 9.5+/-4.3, respectively). CONCLUSION: Our results indicate that rejection in the tilapia-to mouse model follows a pattern similar to other models of discordant islet cell xenotransplantation. PMID- 12640297 TI - Accommodation after lung xenografting from hamster to rat. AB - BACKGROUND: Long-term xenograft survival can be achieved in hamster hearts transplanted into rats treated with cobra venom factor (CVF) and cyclosporine A (CsA). This phenomenon of "accommodation" is associated with expression of protective genes such as bcl-2, bcl-X(L), and heme-oxygenase-1. We examined whether accommodation could be induced in hamster-to-rat lung xenografts and whether the pattern of protective genes is similar to cardiac xenografts. METHODS: We used hamster-to-rat cardiac and lung xenotransplantation models. Cardiac xenotransplants were treated with CVF+CsA and compared with untreated controls. Lung xenotransplants were treated with either CVF+CsA or FK506 and cyclophosphamide (Cp) and compared with untreated controls. All recipients were killed by 21 days after transplantation. We examined graft survival and protein expression of protective genes, and we performed histologic and immunohistologic analyses. RESULTS: Rejection occurred rapidly in untreated rats. CVF+CsA or FK506+Cp treatment significantly influenced graft survival. Eight of 12 CVF+CsA treated heart transplants survived 21 days. Seven of 16 CVF+CsA-treated lung grafts and five of 12 FK506+Cp-treated lung xenografts survived 21 days. We observed significant protein expression of bcl-2, bcl-X(L), and heme-oxygenase-1 in cardiac xenografts treated with CVF+CsA at 2, 14, and 21 days after transplantation, compared with normal hamster hearts. We also observed significant expression of these proteins in lung xenografts treated with either CVF+CsA or FK506+Cp at 21 days after transplantation, compared with normal lungs. CONCLUSIONS: Accommodation may be a general phenomenon for all organs, mediated through protective genes. Induction of accommodation does not require disruption of the complement system. PMID- 12640299 TI - Indefinite islet protection from autoimmune destruction in nonobese diabetic mice by agarose microencapsulation without immunosuppression. AB - BACKGROUND: The recurrence of autoimmunity and allograft rejection act as major barriers to the widespread use of islet transplantation as a cure for type 1 diabetes. The aim of this study was to evaluate the feasibility of immunoisolation by use of an agarose microcapsule to prevent autoimmune recurrence after islet transplantation. METHODS: Highly purified islets were isolated from 6- to 8-week-old prediabetic male nonobese diabetic (NOD) mice and microencapsulated in 5% agarose hydrogel as a semipermeable membrane. Islet function was evaluated by a syngeneic islet transplantation model, in which islets were transplanted into spontaneously diabetic NOD mice. RESULTS: The nonencapsulated islet grafts were destroyed and diabetes recurred within 2 weeks after transplantation in all 12 mice. In contrast, 13 of the 16 mice that underwent transplantation with microencapsulated islets maintained normoglycemia for more than 100 days after islet transplantation. Histologic examination of the nonencapsulated islet grafts showed massive mononuclear cellular infiltration with beta-cell destruction. In contrast, the microencapsulated islets showed well granulated beta cells with no mononuclear cellular infiltration around the microcapsules or in the accompanying blood capillaries between the microcapsules. CONCLUSIONS: Agarose microcapsules were able to completely protect NOD islet isografts from autoimmune destruction in the syngeneic islet transplantation model. PMID- 12640300 TI - Cariporide (HOE-642) improves cardiac allograft preservation in a porcine model of orthotopic heart transplantation. AB - BACKGROUND: Acute graft dysfunction caused by ischemia-reperfusion injury is recognized as a major source of morbidity and mortality following adult heart transplantation. The aim of this study was to determine whether treating the donor and recipient with cariporide, an inhibitor of the sodium-hydrogen exchanger, could reduce ischemia-reperfusion injury. METHODS: A porcine model of donor brain death, hypothermic ischemic preservation, and orthotopic cardiac transplantation was used. Allografts in both the control group (CON, n=6) and treatment group (CAR, n=6) were arrested and stored for 4 hours in the extracellular crystalloid cardioplegia currently used in the clinical transplantation program at our institution. In addition, both the donor and recipient animals in the CAR group received a single intravenous dose of cariporide (2 mg/kg) 15 minutes before harvesting and reperfusion, respectively. RESULTS: The initial rate of troponin I release was significantly lower in recipients of CAR hearts than in recipients of CON hearts (P =0.020). All hearts were weaned successfully from bypass. More CAR hearts were weaned successfully at the first attempt, at 1 hour post-reperfusion, than CON hearts (6 of 6 vs 3 of 6), but this did not achieve statistical significance. Left ventricular contractility (preload recruitable stroke-work relationship) and left ventricular compliance (end-diastolic pressure-volume relationship) were significantly better preserved in CAR hearts than CON hearts (both P <0.0001). CONCLUSIONS: Myocardial injury was reduced, and contractile function was better preserved in allografts that received cariporide, compared with allografts that received conventional preservation alone. PMID- 12640298 TI - Long-term survival of intratesticular porcine islets in nonimmunosuppressed beagles. AB - BACKGROUND: The testis is an immunoprivileged organ, and at 37 degrees C, the intratesticular microenvironment supports the survival of allogeneic islets. The objective of this study was to determine whether the immunoprotection afforded by the intratesticular environment is potent enough to prevent the rejection of xenogeneic porcine islets in a large-animal model. METHODS: A bilateral cryptorchid condition was surgically created in sexually mature beagle dogs. Porcine islets were prepared from neonatal pigs by collagenase digestion and 9 days of culture, after which they were injected into each of the cryptorchid testes. Control dogs received liver subcapsular space transplants of porcine islets and autologous islets. After 100 days, the testes and relevant portions of liver were studied immunohistochemically for the presence of islet tissue. RESULTS: The testicular interstitial space of all dogs contained abundant islet tissue. No evidence of lymphocytic infiltration or inflammation was observed. In contrast, porcine islets transplanted to the liver subcapsular space do not survive, although autologous islets engraft well in that position. This occurs even though the recipient's serum contains preformed cytotoxic antibodies to porcine islets that persist after transplantation. CONCLUSIONS: These results demonstrate that the microenvironment existing within the surgically repositioned intra-abdominal testis supports the survival of xenogeneic tissue. The survival of xenogeneic tissue in the absence of immunosuppression in this large-animal model raises the possibility that xenogeneic porcine islet tissue will also survive in humans if transplanted into a similar environment. PMID- 12640301 TI - Generation of donor hematolymphoid cells after rat-limb composite grafting. AB - BACKGROUND: Composite tissue allografts are unique because they provide the vascularized bone marrow with stroma, which is the supportive microenvironment. In this study, we investigated the beneficial effect of donor-derived bone marrow cells within the long-surviving recipient rats after limb transplantation. METHODS: Green fluorescent protein (GFP) transgenic rats developed for paramount cell marking were donors, and wild Wistar rats were recipients. Orthotopic hind limb transplantation was performed using a microsurgical technique. Tacrolimus (1.0 mg/kg) was intramuscularly injected for 14 days postoperatively. The skin graft from GFP donor onto the GFP recipient was performed as a control. Flow cytometric analyses of recipient peripheral blood and bone marrow were carried out at 4 to 6 days, 18 to 21 days, 6 weeks, and 2, 4, 6, 9, and 12 months after transplantation. RESULTS: The rats that received tacrolimus therapy achieved prolonged composite graft acceptance more than 12 months, whereas GFP skin grafts were rejected at 47 days under the same immunosuppressive protocol. Numerous GFP lymphocytes and granulocytes were detected within the recipient bone marrow for the first 6 weeks post limb transplantation. These cells remained relatively stable for more than 12 months. CONCLUSIONS: The results showed that donor derived hematopoietic stem cells engrafted in recipient bone marrow and differentiated to lymphocytes and granulocytes after limb transplantation. The vascularized bone marrow, transplanted as a part of the hind limb, could have contributed to mixed chimerism and worked as the bone-marrow source in the recipients. PMID- 12640302 TI - Prevention of kidney allograft rejection using anti-CD40 and anti-CD86 in primates. AB - BACKGROUND: Costimulation blockade has been proposed to induce allograft tolerance. We combined an antagonist anti-CD40 monoclonal antibody (mAb) with an antagonist anti-CD86 mAb in a rhesus monkey kidney allograft model. We chose this combination because it leaves CD80-CD152 signaling unimpaired, allowing for the down-regulatory effect of CD152 signaling to take place through this pathway. METHODS: Rhesus monkeys underwent transplantation with a major histocompatibility complex-mismatched kidney. One group of animals received anti-CD40 alone, and a second group received the combination of anti-CD40 and anti-CD86, twice weekly for 56 days. RESULTS: Three animals with low levels of anti-CD40 rejected the transplanted kidney while still receiving treatment. Three animals with high levels of anti-CD40 rejected at days 91, 134, and 217 with signs of chronic rejection. Animals treated with the combination of anti-CD40 and anti-CD86 mAbs rejected their kidneys at days 61, 75, and 78, shortly after cessation of treatment. Two animals were killed on days 71 and 116 with a blocked ureter. These animals developed virtually no signs of tubulitis or infiltration during treatment and no donor-specific alloantibodies. CONCLUSIONS: Both treatment protocols prevented rejection for the duration of the treatment in most animals. Blocking costimulation by anti-CD40 or by anti-CD40 plus anti-CD86 may be an effective method to prevent graft rejection and may obviate the need for other immunosuppressive drugs, especially in the immediate posttransplantation period. PMID- 12640303 TI - Long-term limb allograft survival using anti-CD40L antibody in a murine model. AB - BACKGROUND: Costimulation blockade has been shown to be effective in achieving donor-specific immune unresponsiveness in models of organ transplantation. This study represents the first application of blockade of the CD40 costimulatory pathway to a murine model of limb allotransplantation. METHODS: Eighteen Balb/c mice (H-2K(d)) were randomized to four groups. The control group (n=5) received syngeneic limb transplants from Balb/c donors. The experimental groups were recipients of limb allografts from C57Bl/6 mice (H-2K(b)) and received either no treatment (n=5) or treatment with MR1 (hamster antimouse CD40 ligand monoclonal antibody) 500 microg intraperitoneally (IP) on days 0, 2, 4, 6, 14, 28, and 60 (n=5). A fourth group received myocutaneous allografts from C57Bl/6 donors and the same treatment with MR1 (n=5). RESULTS: Untreated limb allografts were rejected at a mean of 9.6+/-1.1 days postoperatively. MR1-treated limb allografts underwent rejection of the skin component at a mean of 75+/-25 days whereas the musculoskeletal component survived to a mean of 222+/-84 days with two allografts surviving more than 10 months (P<0.001). The MR1-treated myocutaneous allografts were rejected after 16.2+/-2 days. All groups demonstrated acute rejection on histology except the treated limb allograft group, which was more suggestive of a chronic process. No chimerism was detected in this group by flow cytometry. CONCLUSIONS: CD40 costimulatory blockade significantly prolonged limb-allograft survival, and the bone-marrow component may have played an important role. Tolerance was not achieved, and histologic evaluation suggested chronic rejection as a possible cause of allograft loss. PMID- 12640304 TI - Recurrent lupus nephritis in renal transplant recipients revisited: it is not rare. AB - BACKGROUND: Although recurrent lupus nephritis (RLN) after kidney transplantation is reported to be rare (1%-4%), recent studies suggest a higher incidence. The purpose of this study was to determine the incidence of RLN in a large cohort of renal transplant recipients with systemic lupus erythematosus (SLE). METHODS: The records of 54 renal transplant recipients with SLE were reviewed. Thirty-one patients underwent biopsy because of worsening renal function and proteinuria. All biopsy specimens were evaluated by light microscopy, immunofluorescence (IF), and electron microscopy (EM). RESULTS: Among the 50 patients with at least 3 months of follow-up, RLN was present in 15 (52% of patients who underwent biopsy, 30% of total patients): mesangial lupus nephritis (LN) (class II) in eight, focal proliferative LN (class III) in four, and membranous LN (class Vb) in three patients. One patient had graft loss because of RLN (class II) at 10.5 years. The duration of dialysis before transplantation was not different between patients with RLN compared to patients without RLN (P=0.40). Overall patient survival (n=50) was 96% at 1 year and 82% at 5 years, and graft survival was 87% at 1 year and 60% at 5 years. Graft survival was worse in patients who underwent biopsy compared with patients who never underwent biopsy (P<0.01). CONCLUSIONS: RLN is more common than previously reported, but in our series, graft loss because of RLN was rare. Aggressive use of allograft biopsies and morphologic evaluation with IF and EM are important factors in the diagnosis of RLN. The impact of new immunosuppressive agents on the incidence of RLN remains to be seen. PMID- 12640305 TI - Functional antibodies to leukocyte adhesion molecules in antithymocyte globulins. AB - BACKGROUND: Polyclonal antithymocyte globulins (ATG) induce T-cell depletion and functional impairment of nondeleted lymphocytes. Interference of ATG with the main leukocyte surface molecules involved in cellular adhesion and leukocyte endothelium interaction was investigated in the present study. METHODS: In three rabbit ATG, the authors measured antibodies to integrins, beta2-integrin ligands, and chemokine receptors by flow cytometry; chemotactic responses; and down modulation of cell surface expression on lymphocytes, monocytes, and neutrophils. RESULTS: Antibodies to CD11a/CD18 (leukocyte function-associated antigen-1 [LFA 1]) present in ATG induced a dose-dependent down-modulation of cell surface expression of this beta2 integrin on lymphocytes, monocytes, and neutrophils. In contrast, anti-LFA-1 monoclonal antibodies did not induce LFA-1 modulation unless cross-linked by a second antibody. ATG also contained functional antibodies to the beta1 integrin CD49d/CD29 (VLA-4), the alpha4beta7 integrin, CD50, CD54, and CD102 but not to CD62L. ATG were shown to bind to CXCR4 and CCR7 on lymphocytes, CXCR4, and CCR5 on monocytes; to down-modulate cell surface expression of CCR7; and to decrease monocyte chemotactic response to CCL5 (RANTES) and lymphocyte chemotactic response to CCL19 (MIP-3beta). CONCLUSION: These results show that ATG may interfere with leukocyte responses to chemotactic signals but mostly inhibit the expression of integrins required for firm cellular adhesion. The latter property of inhibition is not shared by monoclonal antibodies, and it may contribute to decreasing graft cellular infiltration during acute rejection and possibly after postischemic reperfusion. PMID- 12640306 TI - Complement fragment C4d deposition in peritubular capillaries in acute humoral rejection after ABO blood group-incompatible human kidney transplantation. AB - BACKGROUND: Acute humoral rejection (AHR) is the most important risk factor for early graft loss in ABO-incompatible (ABO-i) kidney transplantation (RTx). The pathogenesis and diagnostic criteria for AHR after ABO-i RTx remain unclear. Complement fragment C4d deposition in peritubular capillaries (PTC), which is a sensitive indicator for activation of the classical complement pathway, was studied to establish the pathologic diagnostic indicator of AHR. METHODS: Forty four graft biopsy specimens from 19 patients with ABO-i living donors were analyzed within 90 days after RTx. Nineteen biopsy specimens with acute rejection after ABO-compatible (ABO-c) living-related RTx were used as controls. Diffuse and bright C4d deposition in PTC was considered significantly positive. RESULTS: All of 8 recipients with AHR showed significantly positive C4d in PTC in the ABO i group, but 9 of 11 recipients without AHR were negative. In the ABO-c RTx group, 16 of 19 recipients were negative for C4d in PTC. The prevalence of C4d in PTC was significantly higher in ABO-i RTx (P<0.05). CONCLUSIONS: C4d deposition is valuable as a specific and sensitive indicator for AHR, even of mild severity, in ABO-i RTx. PMID- 12640307 TI - Erosive enterocolitis in mycophenolate mofetil-treated renal-transplant recipients with persistent afebrile diarrhea. AB - BACKGROUND: Diarrhea is the most frequently reported adverse event in mycophenolate mofetil (MMF)-treated transplant patients. The aim of this study was to explore the gastrointestinal tract in MMF-treated renal transplant recipients with persistent afebrile diarrhea to characterize its nature and etiology. METHODS: Renal transplant recipients with persistent afebrile diarrhea (daily fecal output >200 g) were prospectively investigated for infections, morphologic, and functional (gastrointestinal motility and intestinal absorptive capacity) integrity of the gastrointestinal tract; 26 patients met the inclusion criteria. RESULTS: All but one patient had an erosive enterocolitis. Seventy percent of the patients had malabsorption of nutrients, contributing to the diarrhea. In +/-60%, an infectious origin was demonstrated and successfully treated with antimicrobial agents without changes in immunosuppressive regimen. In +/-40%, no infection occurred, but a Crohn's disease-like pattern of inflammation was noted. These patients also had a less pronounced bile-acid malabsorption but a significant faster colonic transit time, correlating with the trough level of mycophenolic acid (MPA). Cessation of MMF, however, was associated with allograft rejection in one third of these patients. CONCLUSIONS: Persistent afebrile diarrhea in renal transplant recipients is characterized by erosive enterocolitis, which is of infectious origin in +/-60%. In +/-40%, a Crohn's disease-like (entero-)colitis was present. Because reduction or cessation of MMF was the only effective therapy, MPA or one of its metabolites may be suggested as a possible cause. However, reduction or cessation of MMF was associated with an increased risk for rejection. PMID- 12640308 TI - Distinct intragraft response pattern in relation to graft size in liver transplantation. AB - BACKGROUND: The molecular mechanism of small-for-size graft injury remains unclear. The aim of this study is to investigate the gene expression pattern of acute phase response in relation to graft size in a rat-liver transplantation model. METHODS: A rat orthotopic liver transplantation model using 30%, 50%, and whole grafts was used. The graft survival rates and liver morphology were compared among the three groups. Two transcription factors, nuclear factor (NF) kappaB (p65) and early growth response (Egr-1), and their downstream genes were compared. RESULTS: According to the graft size, the rats were grouped as follows: group 1 (n=20), 32% (24-47%); group 2 (n=10), 56% (50-65%); and group 3 (n=10), 104% (89-120%). The 7-day survival rates were 20% (P=0.039 vs. group 2, P=0.000 vs. group 3), 60%, and 100% in groups 1, 2, and 3, respectively. Dilation of hepatic sinusoids and vacuolization of hepatocytes were observed in group 1. Up regulation of Egr-1 and endothelin (ET)-1 and over-expression of nitric oxide synthase (iNOS) was found in group 1, but heme oxygenase (HO)-1 and A20 were down regulated. At 24 hours after reperfusion, the intragraft protein level of heat shock protein (Hsp)-70 was significantly lower in group 1 than that in group 3 (12.4 vs. 17.0 ng/mL, P=0.04). More apoptotic nuclei were found in group 1. CONCLUSIONS: Small-for-size graft injury was related to early over-expression of Egr-1 associated with up-regulation of ET-1 and deterioration of intracellular homeostasis reflected by down-regulation of Hsps and A20. PMID- 12640310 TI - Induction of epidermal fatty acid binding protein in intravascular monocytes of renal allografts. AB - During acute rejection of rat renal allografts, numerous activated monocytes accumulate in the vasculature of the graft. These monocytes seem to be involved in allograft destruction. Proinflammatory and effector functions of monocytes and macrophages can be down-regulated by peroxisome proliferators, which are probably transported in the cytoplasm by fatty acid binding proteins (FABPs). We performed renal transplantation in rats in the Dark Agouti-to-Lewis strain combination. Intravascular graft leukocytes were harvested 4 days posttransplantation. Epidermal (E)-FABP mRNA and protein expression were investigated by reverse transcriptase polymerase chain reaction and immunoblotting, respectively. E-FABP expressing cells were identified by immunofluorescence. After allogeneic transplantation, intravascular graft leukocytes expressed E-FABP mRNA and protein. In isografts, significantly lower expression levels were observed. E FABP protein was detected in monocytes expressing ED1 and in alphabeta-T-cell receptor positive T lymphocytes. E-FABP might regulate monocyte activation and may represent a promising target for a therapeutic intervention in allograft rejection. PMID- 12640309 TI - Migration of mesenchymal stem cells to heart allografts during chronic rejection. AB - BACKGROUND: Mesenchymal stem cells (MSC) are pluripotent progenitors for a variety of cell types, including fibroblasts and muscle cells. Their involvement in the tissue repair of allografts during the development of chronic rejection has been hypothesized, but not yet substantiated, by experimental evidence. METHODS: Rat MSC were isolated from circulation using an aortic pouch allograft as a trapping device. The plasticity of these cells was examined in differentiation cultures. One of the resulting MSC lines was immortalized and transduced to express a marker gene. The -labeled cells were then transferred to F344 rats bearing Lewis (LEW) cardiac allografts to measure their localization and contribution to graft tissue repair. RESULTS: The MSC isolated from circulation exhibited multipotential for differentiation in culture, developing into various lineages including osteoblasts, lipocytes, chondrocytes, myotubes, and fibroblasts. Intravenous engraftment of the -labeled cells into recipients of heart transplant resulted in migration of the beta-gal+ cells into the lesions of chronic rejection in the cardiac grafts and homing of the cells to the bone marrow. The majority of beta-gal+ cells present in the allografts exhibited fibroblast phenotypes, and a small number of the cells expressed desmin, indicative of myocyte differentiation. CONCLUSION: MSC vigorously migrated into the site of allograft rejection. This data suggests that they may be attracted to this site to actively participate in tissue repair during chronic rejection. In addition, given the robust migration, the inhibition of MSC differentiation toward fibroblast progeny and induction toward the myocyte lineage may serve as a new strategy for treatment of chronic rejection and allograft tissue repair. PMID- 12640311 TI - Impact of renal transplantation on small vessel reactivity. AB - BACKGROUND: The function of large arteries is altered after renal transplantation. Whether transplantation also induces agonist-dependent functional changes in small arterial renal and extrarenal vessels has not yet been studied. METHODS: Chronic rejection was induced by grafting Lewis rats with kidneys from Fischer rats (FL). Rats that underwent transplantation were bilaterally nephrectomized. Rats that underwent syngeneic transplantation, uninephrectomized rats, uninephrectomized rats with denervated kidneys or with kidneys made ischemic, and native rats served as controls. All animals were treated with cyclosporine for 10 days. Eighteen weeks after surgery, the reactivity of small arteries (220-270 microm) was tested by myography. RESULTS: Weight gain, glomerular filtration rate, and arterial pressure were similar in all groups, whereas proteinuria was elevated in FL. Only kidneys from FL showed glomerular lesions, tubular atrophy, and vasculopathy. Responsiveness of coronary, mesenteric, and femoral resistance vessels to both constrictor and dilator agonists was similar in transplanted and nontransplanted animals. Resistance vessels obtained from both allogeneically and syngeneically transplanted kidneys were more sensitive to norepinephrine, phenylephrine, angiotensin II, and vasopressin than renal vessels from weight-matched controls. Vasodilation in response to acetylcholine and sodium nitroprusside was mitigated in transplanted versus nontransplanted kidneys. CONCLUSIONS: In rat renal transplantation, renal resistance vessel responsiveness to constrictor or dilator stimuli is altered. Extrarenal small vessel function is not affected. The changes in function of renal resistance vessels are not explained by reduction of nephron mass, denervation, ischemia, or chronic rejection. PMID- 12640312 TI - Adenovirus-mediated expression of human CD55 or CD59 protects adult porcine islets from complement-mediated cell lysis by human serum. AB - BACKGROUND: Protection against complement activation may reduce acute islet damage in pig-to-human islet xenotransplantation. Expression of the human complement regulatory proteins decay-accelerating factor (DAF, CD55) or CD59 was induced on intact adult porcine islets (APIs) by adenoviral transduction. The functional capacity of the transgenes was examined in vitro after exposure to fresh human serum. METHODS: Intact APIs were transduced with adenoviral vectors Ad.hDAF or Ad.hCD59 or a control vector. After 3 days, the islets were trypsin dissociated to a single-cell suspension. A cytotoxicity assay was performed in which the islet cells were incubated with human complement active AB serum. Flow cytometry and immunohistochemistry were used to evaluate transgene expression. RESULTS: APIs could be transduced to express hDAF or hCD59. Flow cytometry analysis of islet single cells revealed that only a fraction of the cells expressed the transgene; immunohistochemical staining of transduced islets demonstrated that mainly cells located in the periphery of the islets were expressing the protein. Cells from nontransduced islets or islets expressing the control protein were sensitive to lysis in human sera (66+/-4.0% and 73+/-3.7% cytotoxicity, respectively). Single cells from islets transduced with hDAF and hCD59 were partially protected from lysis. Islet cells expressing hCD59 were slightly less sensitive to lysis (33+/-3.3%) than cells expressing hDAF (45+/ 3.5%). CONCLUSIONS: These data show that intact pig islets can be transduced to express human regulators of complement activation on the surface and that pig islet cells expressing hDAF or hCD59 are less sensitive to complement-mediated lysis. PMID- 12640313 TI - Apoptosis and necrosis after warm ischemia-reperfusion injury of the pig liver and their inhibition by ONO-1714. AB - BACKGROUND: It is still controversial whether a major mode of cell death during hepatic ischemia-reperfusion (I/R) injuries is apoptosis or necrosis. Moreover, the correlation between these cell deaths and the effects of a novel inducible nitric oxide synthase inhibitor (ONO-1714) has not been studied before. METHODS: Pigs were subjected to 180 min of hepatic warm I/R under extracorporeal circulation. The control group was not administered ONO-1714. In the ONO-1714 group, ONO-1714 was administered 5 min before ischemia at a dose of 0.05 mg/kg through a portal vein catheter. The apoptotic and necrotic changes after reperfusion were examined by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling and hematoxylin-eosin staining. Nitrotyrosine, active caspase 3, and cytochrome c were examined by immunohistochemistry. The plasma NO + NO, aspartate aminotransferase, and lactate dehydrogenase levels were also examined. RESULTS: In the control group, the frequency of apoptotic cells was only 2.6%; nevertheless, that of necrotic cells was 37% at 24 hr after reperfusion. ONO-1714 significantly attenuated apoptosis and necrosis, the expression of nitrotyrosine, and the increases of the plasma aspartate aminotransferase, lactate dehydrogenase, and NO(2)- + NO(3)- levels in the reperfusion phase. CONCLUSIONS: A major mode of cell death during hepatic warm I/R injury was necrosis, and apoptosis was not dominant. These necrotic changes were caused by the excess production of peroxynitrite, and ONO-1714 greatly attenuated I/R injuries as the result of inhibition of the peroxynitrite production. PMID- 12640314 TI - Interindividual variations in constitutive interleukin-10 messenger RNA and protein levels and their association with genetic polymorphisms. AB - BACKGROUND: Genetic variations in the interleukin (IL)-10 gene promoter have been associated with levels of induced production of IL-10, disease susceptibility, and allograft rejection. Small amounts of this cytokine are constitutively produced and are important in maintaining the physiologic function of the cytokine network. In this study, we evaluated the distribution of IL-10 basal levels and its genetic regulation in a healthy Spanish population. METHODS: Polymorphisms at the -1,082, -819, and -512 positions of the IL-10 promoter were analyzed by polymerase chain reaction amplification and hybridization with fluorescent-labeled allele-specific probes in 183 Spanish people. Levels of IL-10 messenger (m)RNA were tested by real-time reverse transcription-polymerase chain reaction in 123 healthy donors. Serum concentrations of IL-10 were measured by a highly sensitive ELISA, whereas protein amounts in lipopolysaccharide culture supernatants were quantified by an in-house ELISA. RESULTS: The frequency of IL 10 promoter alleles and haplotypes in our population showed remarkable differences from other Caucasian populations. Large interindividual variations were found in mRNA and protein constitutive levels of IL-10, which allowed its classification in low and intermediate/high producers. We found statistical differences in mRNA concentration between the polymorphic variant GCC/GCC and the low producer genotypes. The G allele at position -1082 was the most important genetic factor in the regulation of constitutive IL-10 mRNA levels. Similarly, we also found an association of this polymorphic position with serum concentration greater than 2 pg/mL. CONCLUSIONS: Constitutive levels of IL-10 (mRNA and serum protein) displayed remarkable interindividual variations, which are genetically controlled by polymorphic variants at the cytokine gene promoter. PMID- 12640315 TI - Human leukocyte antigens DR and AB and kidney retransplantation. AB - BACKGROUND: Although class II human leukocyte antigen (HLA) DR mismatching has been shown to demonstrate a significant effect on kidney regraft survival, it has not generally been clinically emphasized. METHODS: We examined 2,574 kidney retransplants performed in Southeastern Organ Procurement Foundation centers between January 1988 and December 1997 in which there was ABDR typing on both donor and recipient and pretransplant panel reactive antibody (PRA) data. RESULTS: Cox regression of multiple variables demonstrated that the most important risk factors in descending order were DR mismatching, non-white recipient, female donor, PRA as a continuous variable, and cold ischemia time. Although DR mismatching demonstrated a significant effect in white recipients, the impact was much greater in non-white recipients. In both groups, zero to four AB mismatches demonstrated no significant effect on regraft survival if DR was matched and only a minimal effect when DR was mismatched. The discrepancy of these findings with reports that demonstrate a stepwise decrease in regraft survival on the basis of the total zero to six ABDR mismatches was explained by the fact that the zero to six ABDR mismatches are a combination of AB mismatches with little effect and DR mismatches with a major effect. Regraft survival decreased progressively in association with increasing PRA. CONCLUSIONS: DR matching is critically important in kidney retransplantation. There was no significant difference in survival of zero ABDR mismatched retransplants compared with one to four AB and zero DR mismatched retransplants. On the other hand, kidney graft survival of all one to four AB and zero DR mismatches exceeded that of one or two DR mismatched retransplants. We propose that the association of decreasing regraft survival with increasing PRA reflects undetected sensitization to class II, and possibly class I, antigens. PMID- 12640316 TI - Viral escape and T-cell immunity during ganciclovir treatment of cytomegalovirus infection: case report of a pancreatico-renal transplant recipient. AB - BACKGROUND: Pancreas-kidney transplant recipients are at high risk for cytomegalovirus (CMV) disease despite prophylactic ganciclovir therapy. Because the impact of antiviral therapy on anti-CMV immune reactions is unknown, CMV specific T-cell subsets in primary and recurrent CMV infection were analyzed in a pancreas-kidney transplant case study. METHODS: Major histocompatibility complex class I tetramers were used to detect peripheral CMV pp65-specific CD8 T cells. Intracellular cytokine staining was used to determine the frequency of CMV specific CD4 T cells. Conventional virologic parameters and routine laboratory parameters were monitored. For ganciclovir resistance testing, CMV-UL97 genotyping was performed. RESULTS: Despite prophylactic ganciclovir therapy, primary CMV infection induced in vivo expansion of activated CMV-specific CD8 T cells. Interestingly, viral dissemination during recurrent CMV disease was a result of partially ganciclovir-resistant CMV. Recovery after discontinued ganciclovir treatment was associated with the expansion of CMV-specific CD4 T cells. CONCLUSION: Immunologic monitoring may contribute to clinical management of recurrent CMV disease. PMID- 12640317 TI - Life-threatening veno-occlusive disease after living-related liver transplantation. AB - Veno-occlusive disease (VOD) can develop in association with the administration of cytotoxic chemotherapeutic agents and irradiation. In solid-organ transplant settings, azathioprine has been implicated as a predisposing factor. VOD with fatal outcome occurred in a post liver-transplant recipient who had never been exposed to any agents that have the potential to induce VOD. At onset, the disease manifested clinically as gross ascites and progressive jaundice and was observed after clinically diagnosed acute graft rejection. The disease was confirmed by histologic examinations. Histologic studies of biopsy samples from this patient revealed that most small hepatic veins less than 300 microm in diameter were affected, exhibiting concentric intimal thickening with sparse inflammatory cells. A few of the hepatic veins exhibited active endotheliitis with occasional extension of inflammation to neighboring centrilobular areas. Despite intensified immunosuppression, the observed fibrous obliterative changes were irreversible. Although the cause of VOD in this patient is tentative, the damage to the endothelium, associated with acute rejection, is likely to be attributable. VOD deserves recognition as one of the causes for liver dysfunction and persistent ascites after liver transplantation. PMID- 12640318 TI - Angiotensin-converting enzyme inhibitor-induced isolated visceral angioedema in a liver transplant recipient. AB - Isolated visceral angioedema is an extremely rare complication of angiotensin converting enzyme inhibitors (ACEIs). We report the first known case of ACEI associated visceral angioedema occurring in a liver transplant recipient who presented with acute-onset abdominal pain, nausea, vomiting, diarrhea, radiologic findings of small bowel edema, and ascites. Heightened awareness of the phenomenon of isolated ACEI-associated visceral angioedema is necessary given the increasing use of these medications for treating hypertension related to calcineurin inhibitors and the need to avoid unnecessary surgical or diagnostic interventions in solid-organ transplant recipients. PMID- 12640319 TI - Unusual onset of chronic graft-versus-host disease after adult living-related liver transplantation from a homozygous donor. AB - Graft-versus-host disease (GVHD) is a rare complication that occurs after living related liver transplantation (LRLT). This condition usually occurs early after transplantation and is fatal. Although one-way matching between a human leukocyte antigen (HLA)-homozygous donor and a haploidentical recipient has been shown to be a significant risk factor for the development of acute GVHD, chronic GVHD has not been clarified. A 50-year-old woman underwent LRLT for primary biliary cirrhosis with her 32-year-old HLA-homozygous son as the donor. The patient developed chronic GVHD 114 days posttransplant, presenting with a skin rash only. Her atypical onset included neither fever nor gastrointestinal symptoms. The patient responded to corticosteroid therapy and is now doing well at home. Careful donor selection and HLA matching before LRLT should be performed to prevent GVHD. However, the risks associated with grafts from homozygous donors may be unavoidable at present because of the shortage of cadaveric donors in Japan. PMID- 12640320 TI - Impaired revascularization of transplanted mouse pancreatic islets is chronic and glucose-independent. AB - BACKGROUND: Pancreatic islets are avascular immediately after transplantation and depend on revascularization. Recently, the authors found decreased vascular density in mouse islets 1 month after implantation into nondiabetic recipients. This study investigated possible differences in revascularization between islets implanted into nondiabetic and diabetic recipients, and also evaluated changes in vascular density up to 6 months posttransplantation. METHODS: Islets were syngenically transplanted beneath the renal capsule of normoglycemic or alloxan diabetic C57BL/6 mice. One to 6 months later, the animals were killed and the grafts removed. Histologic slides were prepared and stained with Bandeiraea simplicifolia. RESULTS: The vascular density in all transplanted islets was decreased compared with native islets. There were no differences in the islet graft vascular density between nondiabetic and diabetic animals. No improvement over time occurred. CONCLUSIONS: The vascular density is decreased in islets implanted to cure diabetic recipients. No improvement occurs in transplanted islets after 1 month posttransplantation. PMID- 12640321 TI - [Parasuicide and violence: criminological aspects from a study of 21,314 male army recruits in Switzerland]. AB - Suicidal behaviors are not uncommon, but rarely admitted in the families and in society at large. Violence is a public health and social problem of growing concern. Research on the relationship between violence and suicidal behaviors is scarce for the last decades, perhaps because suicide is no longer considered an offense. The present study is based on a sample of 21,314 valid interviews with 20 year old Swiss men, representing over 70% of this cohort. The questionnaire contained over 900 variables, addressing issues such as previous history of parasuicide, self-reported violent acts and injuries, during childhood and adolescence, as well as over the last year before recruits training. The questionnaire also contains various items about mental health, familial and social conditions and situational factors. We used an ordinal scale for measuring violence and operationalisations of mental disturbances according to theoretical and empirical concepts such as conduct disorder and antisocial personality disorder according to DSM IV, the dissocial syndrome according to Rauchfleisch's (1981) definition; 2,6% of the recruits reported at least one suicide attempt during the last year, one third even multiple attempts. Life-time prevalence at age 20 was 3,9%. Suicide attempters often showed signs of psychiatric disorders, especially for a dissocial (OR=6,9) or an antisocial (OR=5,8) syndrome. Other studies have shown that the rate of suicide in France and Switzerland is very high compared to other European countries. Our study confirms the existence of a real behavioural and existential problem among young Swiss men. Half of the suicide attempters had a history of physical violence against others and 20% were themselves victims of violent acts during the previous year. Suicide attempters who were victims were also more violent than those who were not victimized. A suicide attempt during the previous year is a marker for violent behaviours (OR=2,1) and victimisation (OR=1,9) during this same year. We also found a positive link between multiple attempted suicides and the commission of serious violence. It is concluded that parasuicide among young men is related to other violent acts and should be considered as a positive marker of violent behavior in general and victimisation by crime. It seems that suicide is a transgression of social and cultural values because of the amount of aggression behind it, even if it is not a criminal offense. Suicide attempts of young men should receive more attention by professionals in the field of medicine, psychology and social work. Treatment of these men not only prevents an individual mental health problem but also contributes to crime prevention at the community level. Criminological theories could inspire further studies in suicidology, in order to improve the multidisciplinary knowledge of the phenomenology and etiology of parasuicide. In the same way, criminologists as policy makers should attach more attention to parasuicide given its many theoretical and practical connections with crime. PMID- 12640322 TI - [Cannabis and schizophrenia: demographic and clinical correlates]. AB - The high prevalence of psychoactive substance abuse or dependence among schizophrenic patients has now been well established. Mueser et al. stressed the need to assess the abuse of specific classes of substances and analyse the data accordingly. The objective of this study was to compare the socio-demographic correlates and the clinical features in a group of schizophrenic patients with a lifetime cannabis abuse or dependence according to the DSM III-R with a group of schizophrenic patients who had never presented any abuse or dependence. SUBJECTS AND METHODS: The study included 124 subjects with diagnoses of schizophrenia or schizoaffective disorders according to the DSM III-R. Inclusion criteria for participation in the study were age 18 years or older and willingness to provide consent to participate in the study. The inpatients were evaluated when their condition was stabilised. Assessment tools were the psychoactive substance use disorder section of the Composite International Diagnostic Interview (CIDI), the Positive and Negative Syndrome Scale (PANSS), the Global Assessment of Functioning Scale (GAF). Subjects with cannabis abuse or dependence during their lifetime were compared with subjects without abuse or dependence, using chi(2) test for categorical variables and analyses of covariance (ANCOVA) for quantitative variables. RESULTS: Forty-nine subjects (42,6%) presented lifetime abuse or dependence on one or more substances. Since 19 patients with alcohol, stimulant, sedative or opiate abuse or dependence were excluded, the study finally included 96 subjects including a first group of schizophrenic patients with cannabis abuse (n=6) or dependence (n=24) and a second group without any psychoactive substance abuse (n=66). Thirteen (11.3%) patients presented cannabis abuse or dependence within the 6 months prior to the assessment. The mean SD age of onset of cannabis abuse or dependence was 19.6 +/- 3.0 years. Cannabis abuse/dependence preceded the first psychiatric treatment in 70% of the subjects (n=21). 83.3% of the schizophrenic patients with cannabis abuse or dependence were male (n=25) compared to 62.1% in the group without substance abuse (n=41) (chi(2)=4.32, df=1, p=0.04). Schizophrenic patients with cannabis abuse were significantly younger (mean age: 28.9 +/- 6.3 vs 37.0 +/- 12.7, ANCOVA, F=7.2, df=1,96 p=0.009). There was no significant difference between the two groups for marital status, (chi(2)=5.34, df=2, p=0.07), level of education, (chi(2)=0.93, df=2, p=0.62) professional status, (chi(2)=8.7, df=5, p=0.11), on PANSS total score (ANCOVA, F=0.42, df=1,93, p=0.52), GAF score (ANCOVA, F=0.06, df=1,92, p=0.80), mean number of hospitalizations (ANCOVA, F=3.25, df=1,85, p=0.08), mean age of first psychiatric contact (ANCOVA, F=0.74, df=1,93, p=0.39), and neuroleptic dosages (ANCOVA, F=0.03, df=1,90, p=0.87). In contrast, the total duration of hospitalization was significantly longer for the group with cannabis abuse. Patients with cannabis abuse were more likely to have an history of suicide attempts than subjects without substance abuse (chi(2)=11.52, df=1, p=0.0007). DISCUSSION: The prevalence rates for substance abuse and the socio demographic characteristics of the population of our study are consistent with findings of previous studies. Male gender and age were significantly related to history of cannabis abuse or dependence. Cannabis abuse frequently preceded the onset of psychiatric treatment. However, both schizophrenia and substance abuse tend to develop gradually, with no clear demarcation for the onset of schizophrenia. The absence of any link between the scores for the subscales of the PANSS and cannabis abuse, both in our study and in some retrospective previous studies, is not suggestive of cannabis abuse as a self-medication of positive or negative symptoms of schizophrenia. Self-medication could concern other symptoms, such as cognitive deficits. In addition, the hypothesis of self medication has especially been suggested in cocaine abuse or dependence. Some limitations to this study can be discussed. First, although the recruitment was systematic and done in a public mental health service, the patients of our study are not necessarily representative of all schizophrenic patients. Secondly, as in any retrospective study, the prevalence of lifetime substance abuse may have been under-estimated. Urinary toxicology tests may have been able to improve the sensitivity of the diagnosis of recent substance abuse, but structured interviews are more appropriate for the diagnosis of lifetime substance abuse in schizophrenic patients than urinary toxicology tests. CONCLUSION: The socio demographic characteristics of cannabis abuse or dependence in schizophrenia are similar to those found in general population. Cannabis using schizophrenic patients were more likely to be younger and male than non users. The duration of hospitalization was significantly longer for the group with cannabis abuse. Prevalence of suicide attempts in schizophrenia is closely correlated to cannabis abuse. PMID- 12640323 TI - [Post-traumatic stress disorder (PTSD): the syndrome with multiple faces]. AB - We choose to discuss from the PTSD's point of view because this diagnostic reference is commonly used. We wish outline its restrictive sight which could prevent the professional from having a diagnosis of PTSD. We don't want to say there is a PTSD everywhere but it appears to us that a traumatic reading can be a precious advantage for the clinician to establish a real therapeutic relation with some patients. Post-traumatic syndrome differs from the majority of other diagnostic categories as it includes in its criteria the presumptive cause of the trauma (criterion A). In the case that this syndrome originates in war experiences, the presumed cause presents itself as an exceptional event overcoming the individual's resources. The notion of war traumatisation has been extended to other events such as catastrophes, physical attacks, rapes, child and wife battering, and sexual abuses. But the events which cause PTSD (Post Traumatic Stress Disorder) are significantly more numerous. It can be seen that medical events such as giving birth, miscarriage, heart attack, cancer, or hospitalisation following resuscitation may give rise to PTSD. Further, people experiencing prolonged periods of distress may equally develop a post-traumatic syndrome without any particular event having occurred to surpass their defences. It's the case of the Prolonged Duress Stress Disorder (PDSD). The series of discontinuous stress "waste" the psychic balance and may give rise, at one moment, to posttraumatic symptoms described in DSM, without any specific stressful event. The existence of criterion A is therefore not a necessary prerequisite in establishing a diagnosis of PTSD. It is, in fact, very difficult to predict which events could cause a PTSD, and this, especially, as the subjective aspects count at least as much as the objective aspects. The clinician should have to carefully explore how the patient experienced the event or, how he apprehended the event itself and it's outcome, if he wants get the traumatic range of a life event. The feeling of deep distress, the feeling of being trapped, the loss of control, the collapse of basic beliefs, the feeling that one's life is in jeopardy, that the physical integrity is (really or in one's imagination) threatened, the feeling of helplessness, are quite as much clues for a possible PTSD which hides behind others clinical manifestations either psychological or somatic. Furthermore, the "pure" form described in the DSM and grouping together three further criteria (reliving events, avoiding stimuli associated with the trauma, hyper-reactivity) is extremely rare in the chronic form. An untreated post-traumatic syndrome evolves with time and may present, initially, with very different pathological symptoms giving rise to equally varied diagnoses. Different etiopathogenic models propose to account for the PTSD 's heterogeneous appearance and instability with time. The comorbidity concept sees the PTSD as an independent entity other independent pathologies coexist with. The typologic concept suggests that the PTSD is an independent entity which shows different clinical appearances based on symptomatic descriptions. The "cascade" concept suggests to see the PTSD as an independent entity which offers, with time, different symptomatic appearances, in evolution, because of events caused by after effects, in different areas of the PTSD itself. All of these concepts outline the transnosologic appearance of the PTSD which makes it hardly recognizable. The "chronic" syndrome is rarely diagnosed forming a real challenge to prevention. In effect, the present authors insist on the crucial nature of early detection of PTSD since the greater the time elapsed the more difficult it becomes due to the evolutionary aspect of the syndrome, which initially has more readily recognizable symptoms. The consequences of an unrecognised PTSD are serious and affect both the individual and his immediate family and friends, contributing further to the aggravation of the problems. When a PTSD is diagnosed, it can allow the clinician to further a more global care which will help the patient to get a better recovery. With patients who suffered an infarct, the treatment of PTSD which prevents their recovery will help to go back to the way they lived before the event. It has been showed how important could be the PTSD detection on the severe burned people's pain control. Thus it seems to be crucial for the clinician to keep this diagnosis in mind alongside any other. PMID- 12640324 TI - [Dimensional personality assessment of the members of the French junior national team of road cycling]. AB - The high-standard sport practice requires an optimum functioning level of the individual in all its physiological systems as a whole. If the physical program training progressed much these last years, the techniques of mental preparations are still very heterogeneous and are not based on any validated procedures, based only on individual practices. Nevertheless, the majority of athletes and trainers stress the major importance of "mental" in the realization of performances. One of the obstacles in the mental training of the athletes is the difficulty in finding tools making it possible to evaluate and measure the psychic state of the individual and its mode of coping and adjustment, apart from any psychopathology. Few studies have been carried out on applicability of the personality questionnaires derived from the current dimensional models in these populations. Our study aimed to describe the personality of the members of the french junior team of road cycling, using the Temperament and Character Inventory (TCI) developed by Cloninger to explore the seven dimensions of his psychobiologic model of personality. In this model, four dimensions are temperament factors, and three are character factors. The three main dimensions of temperament are Novelty Seeking (NS), ie the tendency towards excitement in response to novel or rewarding stimuli, Harm Avoidance (HA) hypothesized to represent the tendency to respond intensely to signals of adverse stimuli, and Reward Dependence reflecting the tendency to respond intensely to signals of reward and to maintain behavior previously associated with reward. These personality traits are hypothetically related to underlying neurotransmitter systems (especially NS to dopaminergic function and HA to serotonergic function). The two main dimensions of character are Self-Directedness (SD) and Cooperativeness (C), measuring maturity traits respectively concerning individual and social adaptation. In this study, we used a computerized version of the TCI in a sample of 18 racing cyclists, ie all the members of the french junior national team. Their results were compared to those of 26 male, young, community subjects, derived from the french normative validation sample of the TCI. Mean comparisons were performed with the Mann Whitney test, with Bonferroni correction for multiple comparisons. A significant difference between subjects and controls was obtained for Reward Dependence (p<0.0001), attachment RD3 (p<0.0001), and dependence RD4 (p<0.0025). No significant difference appeared between both groups for the other temperament or character dimensions. Based on temperamental Cloninger typology, the type "passionate" appeared to be the most frequent in this sample (55.6%). These results suggested that the TCI can be easily and usefully used in athletes, and that the personality profile of young competitive cyclists is not abnormal except a high level of reward dependence. Numerous applications can be derived from this use: the development of specific and personalized mental training, the screening and prevention of psychological vulnerability. PMID- 12640325 TI - [Self-esteem: a comparison study between eating disorders and social phobia]. AB - Eating disorder patients evidenced very often a low self-esteem. Self-esteem in eating disorder patients is excessively based on body dissatisfaction. In eating disorders there seems to be a link between body image dissatisfaction and social anxiety. We hypothesised: self-esteem would be as low in eating disorder patients as in social phobia patients; self-esteem would be lower in eating disorder patients with social phobia than in patients with social phobia alone; self esteem would be lower in eating disorder patients with depressive cognitions than in social phobia patients with depressive cognitions; self-esteem could have different characteristics in the two disorders; self-esteem would be as low in anorexia as in bulimia; 103 eating disorder patients (33 restrictive anorectics, 34 anorectics-bulimics, 36 bulimics) and 26 social phobia patients diagnosed according to DSM IV and ICD-10 criteria have been investigated by the Self-Esteem Inventory of Coopersmith, the Assertiveness Schedule of Rathus, the Fear Survey Schedule of Wolpe (FSS III) and the Beck Depression Inventory (BDI). Patients were free of medication and presented no episode of major depression according to DSM IV criteria. Evaluations took place before any psychotherapy. Self-esteem in eating disorder patients is reduced at the same level as in social phobia patients; 86.1% of the total sample and 84.5% of the eating disorder patients have a very low self-esteem (score 33 in the SEI). Eating disorder patients have significantly higher scores in the Social (p=0.016) and Professional (p=0.0225) sub-scales of the SEI than social phobia patients. Eating disorder patients show higher scores on the Assertiveness Schedule of Rathus (p=0.0013) than social phobia patients. Eating disorder patients disclose higher scores on the BDI (p=0.0003) but eating disorder patients with depressive cognitions do not differ from social phobia patients with depressive cognitions in the level of self esteem. The FSS III scores are significantly lower in eating disorder patients (p<0.0001). There is a difference in the nature of the deficit of self-esteem between the two patient populations. Self-esteem is not influenced by the Body Mass Index (BMI) and is identically reduced in all groups of eating disorder patients. Whereas eating disorder patients have the same complaints compared to social phobia, they differ significantly from social phobia patients in their characteristics of social phobia and self-esteem. PMID- 12640326 TI - [The Tuaregs addiction to tea, to smokeless tobacco and to milk: ethnological and clinical approach]. AB - Some rituals about a regular consumption of tea, smokeless tobacco (chewing) and milk are described by one of the authors at the time of his anthropological investigation among the Tuaregs of Timbuktu's region (Mali). He carries out some ethnographical and clinical materials which highlight the dependence to these substances and the role of their psychostimulant and anorexigene effects in a society much ritualised. The subject of this article appears original in the literature which approaches more the dependence to coffee than tea, to cigarettes than to chewing tobacco. The observation of daily life of a tuareg encampment shows a ritual consumption of tea at four time a day. The motivations of the Tuaregs are the increase of vigilance and performance with that psychostimulant substance. They describe an intoxication syndrome related to caffeineism, observed among European tourists. The Tuaregs are aware of their addiction to tea and distinguish psychological dependence from physical dependence. The psychological dependence corresponds to a powerful desire to drink tea at ritual moments, while the physical dependence appears at waking-up and when the time of preparing this beverage is too late. The Tuaregs describe also a phenomenon of loss tolerance after an abstinence period. In spite of the maraboutic prohibition to drink tea, which diverts Tuaregs of their religious practice, they defy this ban from the waking-up to take that infusion before the matinal prayer. That addiction appears also in the identity of the Tuaregs who are called "the sons of tea". The consumption of chewing tobacco, mixed with ash, rhythms the daily life. The mean number of chewing is about fifteen by day; every chewing last 30 minutes. The first chewing of the day occurs 15 minutes after waking-up. The Tuaregs use tobacco in order to get relaxation and vigilance. They suggest intoxication symptoms and especially a withdrawal syndrome which appears at the waking-up or after an important interval between chewing. The authors raise the idea about the dependence to this type of tobacco, consistent with the Anglo Saxon literature of the 80th which tried to implement scales and criteria as to assess the dependence to smokeless tobacco. The Tuaregs could be more addicted than American consumers in regard to american studies: they use more chewing a day and they can't refrain from chewing at the waking-up. Empirical addition of plant ash, made up of hydroxide of calcium, may act a role in pharmacokinetic by alkalinising the pH. It could increase the absorption of nicotine through the mouth mucus membrane. The authors raise the idea about the dependence to the milk, much consumed and ritualised among those nomadic breeders. They rely on the observation of a withdrawal syndrome clearly identified in the tuareg medical nosography. These regular consumptions integrate the daily life within other rituals. Tea and tobacco facilitate certain motor stimulation, a struggle against hunger and some relaxation regarding an hostile environment over climatological, ecological and economical plan. The brutal and unexpected occurring of one of those rituals disrupt, indeed invert, the usual order of social rituals. Those social and religious disruptions materialise the pathological effect of that double dependence to nicotine and caffeine. That one is called by a term which translate its subjective and social appearance, reflecting so the interaction between man, environment and psychoactive substance. This article highlight the importance of cultural factors in the etiopathogeny of poly-dependence among Tuareg subjects. The question about the diagnostic of the dependence in the DSM IV and the CIM-10 is raised. The DSM IV could be completed because it doesn't evoke addiction to caffeine of tea such like it is consumed in West actually. That hermeneutic approach, including anthropological observations and clinical investigations, allow to understand that addiction to psychoactive substances among Tuareg subjects is consistent with their survival in hostile environments. PMID- 12640327 TI - [Lipids, depression and suicide]. AB - Polyunsatured fatty acids are made out of a hydrocarbonated chain of variable length with several double bonds. The position of the first double bond (omega) differentiates polyunsatured omega 3 fatty acids (for example: alpha-linolenic acid or alpha-LNA) and polyunsatured omega 6 fatty acids (for example: linoleic acid or LA). These two classes of fatty acids are said to be essential because they cannot be synthetised by the organism and have to be taken from alimentation. The omega 3 are present in linseed oil, nuts, soya beans, wheat and cold water fish whereas omega 6 are present in maize, sunflower and sesame oil. Fatty acids are part of phospholipids and, consequently, of all biological membranes. The membrane fluidity, of crucial importance for its functioning, depends on its lipidic components. Phospholipids composed of chains of polyunsatured fatty acids increase the membrane fluidity because, by bending some chains, double bonds prevent them from compacting themselves perfectly. Membrane fluidity is also determined by the phospholipids/free cholesterol ratio, as cholesterol increases membrane viscosity. A diet based on a high proportion of essential polyunsatured fatty acids (fluid) would allow a higher incorporation of cholesterol (rigid) in the membranes to balance their fluidity, which would contribute to lower blood cholesterol levels. Brain membranes have a very high content in essential polyunsatured fatty acids for which they depend on alimentation. Any dietary lack of essential polyunsatured fatty acids has consequences on cerebral development, modifying the activity of enzymes of the cerebral membranes and decreasing efficiency in learning tasks. EPIDEMIOLOGICAL DATA: The prevalence of depression seems to increase continuously since the beginning of the century. Though different factors most probably contribute to this evolution, it has been suggested that it could be related to an evolution of alimentary patterns in the Western world, in which polyunsatured omega 3 fatty acids contained in fish, game and vegetables have been largely replaced by polyunsatured omega 6 fatty acids of cereal oils. Some epidemiological data support the hypothesis of a relation between lower depression and/or suicide rates and a higher consumption of fish. These data do not however prove a relation of causality. CHOLESTEROL AND DEPRESSION: Several cohort studies (on nondepressed subjects) have assessed the relationship between plasma cholesterol and depressive symptoms with contradictory results. Though some results found a significant relationship between a decrease of total cholesterol and high scores of depression, some other did not. Studies among patients suffering from major depression signalled more constantly an association between low cholesterol and major depression. Besides, some trials showed that clinical recovery may be associated with a significant increase of total cholesterol. CHOLESTEROL AND SUICIDAL BEHAVIOR: The hypothesis that a low cholesterol level may represent a suicidal risk factor was discovered accidentally following a series of epidemiological studies which revealed an increase of the suicidal risk among subjects with a low cholesterol level. Though some contradictory studies do exist, this relationship has been confirmed by several subsequent cohort studies. These findings have challenged the vast public health programs aimed at promoting the decrease of cholesterol, and even suggested to suspend the administration of lipid lowering drugs. Recent clinical studies on populations treated with lipid lowering drugs showed nevertheless a lack of significant increase of mortality, either by suicide or accident. In addition, several controlled studies among psychiatric patients revealed a decrease of the concentrations of plasma cholesterol among patients who had attempted suicide in comparison with other patients. POLYUNSATURATED FATTY ACID AND DEPRESSION: In major depression, all studies revealed a significant decrease of the polyunsaturated omega 3 fatty acids and/or an increase of the omega 6/omega 3 ratio in plasma and/or in the membranes of the red cells. In addition, two studies found a higher severity of depression when the level of polyunsaturated omega 3 fatty acids or the ratio omega 3/omega 6 was low. Parallel to these modifications, other biochemical perturbations have been reported in major depression, particularly an activation of the inflammatory response system, resulting in an increase of the pro inflammatory cytokines (interleukins: IL-1b, IL-6 and interferon g) and eicosanoids (among others, prostaglandin E2) in the blood and the CSF of depressed patients. These substances cause a peroxidation and, consequently a catabolism of membrane phospholipids, among others those containing polyunsaturated fatty acids. The cytokines and eicosanoids derive from polyunsaturated fatty acids and have opposite physiological functions according to their omega 3 or omega 6 precursor. Arachidonic acid (omega 6) is, among others, precursor of pro-inflammatory prostaglandin E2 (PGE2), whereas polyunsaturated omega 3 fatty acids inhibit the formation of PGE2. It has been shown that a dietary increase of polyunsaturated omega 3 fatty acids reduced strongly the production of IL-1 beta, IL-2, IL-6 and TNF-alpha (tumor necrosis factor-alpha). In contrast, diets with a higher supply of linoleic acid (omega 6) increased significantly the production of pro-inflammatory cytokines, like TNF alpha. Therefore, polyunsaturated omega 3 fatty acids could be associated at different levels in the pathophysiology of major depression, on the one hand through their role in the membrane fluidity which influences diverse steps of neurotransmission and, on the other hand, through their function as precursor of pro-inflammatory cytokines and eicosanoids disturbing neurotransmission. In addition, antidepressants could exhibit an immunoregulating effect by reducing the release of pro-inflammatory cytokines, by increasing the release of endogenous antagonists of pro-inflammatory cytokines like IL-10 and, finally, by acting like inhibitors of cyclo-oxygenase. THERAPEUTIC USE OF FATTY ACIDS: Data available concerning the administration of supplements of DHA (docosahexanoic acid) or other polyunsaturated fatty acids omega 3 are limited. In a double blind placebo-controlled study on 30 patients with bipolar disorder, the addition of polyunsaturated omega 3 fatty acids was associated with a longer period of remission. Moreover, nearly all the other prognosis measures were better in the omega 3 group. Very recently, a controlled trial showed the benefits of adding an omega 3 fatty acid, eicosopentanoic acid, among depressed patients. After 4 weeks, six of the 10 patients receiving the fatty acid were considered as responders in comparison with only one of the ten patients receiving placebo. CONCLUSIONS: Some epidemiological, experimental and clinical data favour the hypothesis that polyunsaturated fatty acids could play a role in the pathogenesis and/or the treatment of depression. More studies however are needed in order to better precise the actual implication of those biochemical factors among the various aspects of depressive illness. PMID- 12640328 TI - [Criteria defining antimanic drugs (psychopharmacological specificity and/or nonspecificity?)]. AB - The question as to whether specific antimanic drugs differ in their action profile from nonspecific drugs is addressed in regard to symptomatic, nosographic, regulatory and physiopathological issues. Results from clinical studies have shown that mood stabilizers and typical neuroleptics differ as regards improvement of manic symptoms: the former appear to act more evenly on all symptoms of mania, showing a more total normalization of affect, ideation and behaviour whereas the latter tend to sedate patients or to cause a psychomotor retardation, leaving the core manic symptoms unaffected. This has been many times underlined, in particular for lithium, notwithstanding the fact that rating scales employed in clinical trials have often been charged to fall far short of being sensitive enough to pick up the qualitative changes in manic psychopathology. Antimanic drugs may also be more or less specific in their capacities to treat all facets of the manic episode (psychotic, depressive, irritable) whatever the bipolar subtype (bipolar I, II, rapid and non-rapid cycling, secondary bipolar disorder) or the disease stage (early and late episodes). In this respect divalproate seems to have a broader spectrum of efficacy than other available agents. Newer antipsychotics such as olanzapine are promising too. From a regulatory point of view, the current European requirements for a specific antimanic drug are more stringent than the US requirements of the Food and Drug Administration (FDA). Efficacy must be demonstrated in short-term studies showing an effect in acute mania; moreover it has to be shown that efficacy is to be maintained during the episode. So far, three armed randomized controlled trials are required, in which the test product is compared both with placebo and with a standard treatment. A possible design is a comparison of test product, placebo and active control for 3 weeks followed by a two-arm phase for the remaining 9 weeks, comparing only test product and active control. In addition, a specific antimanic has to demonstrate that it does not cause switching to depression. As regards physiopathology, integrative models of bipolar disorder, ie kindling and behavioural sensitization, offer an exciting perspective on the specificity issue; agents active in these models initialize a cascade of intracellular signaling that leads to changes in the expression of immediate early genes as well as late effector genes in corticolimbic structures: the former may contribute to acute symptomatology whereas the latter give rise to neuroanatomical reorganization which could underlie more stable changes in mood and cognition. Due to their action on intracellular messaging systems, dopamine D(1) receptors, serotonin 5HT(1a) or 5HT(2a) receptors, especially in orbitofrontal circuit, antimanic agents may exhibit a more specific activity than other drugs. This specificity could concern a whole spectrum of bipolarity which might be characterized by impulsivity. PMID- 12640329 TI - [Efficacy of a non blind placebo prescription]. AB - The aim of our open clinical trial was to determine the efficacy and the tolerance of a blue coloured placebo in moderated anxious patients (Hamilton score below 15) and a red one for tired patients without DSM IV criteria of major depressive disorder. All the patients knew that treatment was a placebo and so, had no pharmacological effect. The trial run over seven days. All patients were considered as responders if their initial symptoms were reduced more than 50%. At the end of the week, the final clinical evaluation showed that 18 patients about 34 were in good condition (10 anxious patients, 7 women and 3 men, and 8 tired patients, 4 women and 4 men). The anxious score of Hamilton scale was reduced of 63%; 16 of the 18 responders were absolutely sure that the treatment was usefull. Four patients were obliged to stop their treatment because side effects: insomnia, tiredness and sleepiness, gastric pain and itching of fore arm. PMID- 12640330 TI - [Catatonia de novo, report on a case: immediate vital prognosis and psychiatric prognosis in longer term]. AB - We report on the case of a 20 year old woman with no previous psychiatric history, who displayed a first episode of catatonia with acute onset. Symptoms started plainly with sudden general impairment, intense asthenia, headache, abdominal pain and confusion. After 48 hours, the patient was first admitted to an emergency unit and transferred to an internal medicine ward afterwards. She kept confused. Her behaviour was bizarre with permanent swinging of pelvis, mannerism, answers off the point and increasingly poor. The general clinical examination was normal, except for the presence of a regular tachycardia (120 bpm). The paraclinical investigations also showed normal: biology, EEG, CT Scan, lumbar puncture. Confusion persisted. The patient remained stuporous, with fixed gazing and listening-like attitudes. She managed to eat and move with the help of nurses but remained bedridden. The neurological examination showed hypokinaesia, extended hypotonia, sweating, urinary incontinence, bilateral sharp reflexes with no Babinski's sign and an inexhaustible nasoorbicular reflex. The patient was mute and contrary, actively closed her eyes, but responded occasionally to simple instructions. For short moments, she suddenly engaged in inappropriate behaviors (wandering around) while connecting back to her environment answering the telephone and talking to her parents. The patient's temperature rose twice in the first days but with no specific etiology found. During the first 8 days of hospitalization, an antipsychotic treatment was administered: haloperidol 10 mg per os daily and cyamemazine 37.5 mg i.m. daily. Despite these medications, the patient worsened and was transferred to our psychiatric unit in order to manage this catatonic picture with rapid onset for which no organic etiology was found. On admission, the patient was stuporous, immobile, unresponsive to any instruction, with catalepsy, maintenance of postures, severe negativism and refusal to eat. A first treatment by benzodiazepine (clorazepate 20 mg i.v.) did not lead to any improvement. The organic investigations were completed with cerebral MRI and the ruling out of a Wilson's disease. Convulsive therapy was then decided. It proved dramatically effective from the first attempt; 4 shocks were carried out before the patient's relatives ask for her discharge from hospital. The patient revealed she had experienced low delirium during her catatonic state. The clinical picture that followed showed retardation with anxiety. She was scared with fear both for the other patients and the nursing team. She kept distant and expressed few affects. The treatment at the time of discharge was olanzapine 10 mg per os. She was discharged with a diagnosis of catatonia but with no specific psychiatric etiological diagnosis associated. She discontinued her follow-up a few weeks later. After one year, we had no information about her. Catatonia has now become rare but remains a problem for clinicians. We reviewed data concerning short term vital prognosis and psychiatric long term prognosis in catatonia. Lethal catatonia is associated with acute onset, both marked psychomotor and neurovegetative symptoms. In the light of literature, there is no proband clinical criterion during the episode that is of relevant diagnostic value to ascertain the psychiatric etiology. PMID- 12640331 TI - Heparin stimulates production of bFGF and TGF-beta 1 by human normal, keloid, and fetal dermal fibroblasts. AB - BACKGROUND: Heparin decreases dermal fibroblast proliferation and collagen production according to several studies. Heparin may mediate these effects by altering the levels of growth factors such as basic fibroblast growth factor (bFGF) and transforming growth factor-beta 1 (TGF-b1). This study sought to delineate the effect of heparin on proliferation and bFGF and TGF-b1 production by human normal, keloid, and fetal dermal fibroblasts. MATERIAL/METHODS: Human normal, keloid, and fetal dermal fibroblasts were propagated in a serum-free in vitro model, with exposure to 0 microg/ml, 50 microg/ml, 300 microg/ml, or 600 microg/ml heparin for 0, 24, 72, or 96 hours. Cell counts were determined by phase contrast microscopy. Levels of bFGF and TGF-b1 in the supernatants were determined by enzyme-linked immunosorbant assay (ELISA). RESULTS: Heparin inhibited keloid and fetal fibroblast proliferation. All doses of heparin significantly stimulated production of bFGF by normal (341% to 1137% increase), keloid (237% to 1955% increase), and fetal fibroblasts (292% to 1866% increase) at all time points (p<0.05). Heparin (300 microg/ml and 600 microg/ml) also stimulated production of TGF-b1 by normal (56% to 75%), keloid (105% to 269%), and fetal fibroblasts (25% to 57%), with statistical significance (p<0.05) at various time points. 600 microg/ml heparin generally caused the greatest increase in growth factor levels. CONCLUSIONS: Heparin inhibits proliferation by keloid and fetal fibroblasts and significantly stimulates production of bFGF and TGF-b1 by normal, keloid, and fetal dermal fibroblasts. These effects of heparin on dermal fibroblasts may have implications for wound healing in vivo. PMID- 12640332 TI - A mouse model for studying rapid intraoperative methods of skin closure and wound healing. AB - BACKGROUND: We re-examine the adequacy of an albino (hairless) mouse dorsal distally based skin flap model by determining elastic modulus and by assessing viability after application of rapid intermittent stretch to facilitate wound closure. MATERIAL/METHODS: Modulus of elasticity was determined with materials testing. Viability was accessed by laser Doppler flowmetry and by quantifying marginal tissue necrosis (mm2) at 5 days. Effective length gained and tension relieved were measured by subjecting flaps to simple undermining plus 3-10 g of initial tension (linear stretch) using a skin hook. Immediate hemispherical stretch was applied with an implanted subcutaneous silicone balloon (Rapid Intraoperative Tissue Expansion or RITE) at 1 or 2 cycles of 3-8 min and 6-12 ml for each cycle. Flaps subjected to simple and extensive undermining with or without application of initial linear stretch served as postoperative controls. RESULTS: The elastic modulus of 9.85 (1.02) N/mm2 approximated the skin of the human face. Flowmetry demonstrated that all flaps were adequately perfused. Stretching significantly decreased flap tension and increased flap length over simple undermining alone. It drastically improved flap viability by provoking a 50-75% decline in 5-day necrosis. The benefits were dependent upon the initial linear tension applied, but independent of the timing, cycling, and volume selected for RITE and the flap dimensions at closure. CONCLUSIONS: Results demonstrate that the albino (hairless) mouse dorsal distally based skin flap is a reliable model for studying wound closure and that rapid intermittent stretching in this model facilitates closure and wound healing. PMID- 12640333 TI - Anomalous traversement of the suprascapular artery through the suprascapular notch: a possible mechanism for undiagnosed shoulder pain? AB - BACKGROUND: The literature is deficient in studies aimed at quantifying the occurrence of a suprascapular artery traversing the suprascapular notch with the suprascapular nerve. MATERIAL/METHODS: We have performed a cadaveric study using 60 adult specimens (120 sides). Thirty-eight specimens were male and twenty-two were female. RESULTS: Our findings conclude that in this group, the suprascapular artery travels with the suprascapular nerve through the suprascapular notch in 2.5% of specimen sides. Both specimens in whom this occurred were male. CONCLUSIONS: We hypothesize that this anatomic anomaly may also constitute a possible unrecognized mechanism in which shoulder pain is generated. Further studies focused at various races and the presence of this peculiarity are now needed to confirm our findings. In addition, unexplained chronic shoulder pain may be explored with vascular studies aimed at identifying the course of the suprascapular artery. PMID- 12640334 TI - A human whole blood model of LPS-mediated suppression of T cell activation. AB - BACKGROUND: Lipopolysaccharide (LPS) is the main initiator of the early signaling events leading to sepsis caused by Gram-negative bacteria. Late stages of sepsis are associated with impairments of T lymphocyte function, a condition associated with nosocomial infection and poor outcome. The molecular basis for septic immunosuppression is not fully understood. MATERIAL/METHODS: Human whole blood was incubated ex vivo with purified LPS. Cytokine responses and T cell proliferation were assessed, and the role of cyclic 3',5'-adenosine monophosphate (cAMP) in T cell suppression by LPS was studied using a cAMP-antagonist (Rp-8-Br cAMPS). RESULTS: Adding LPS (0.01 to 10 microg/ml) to human blood ex vivo caused a release of prostaglandin E2 (PGE2) in a concentration- and time-dependent manner, with maximal levels of PGE2 obtained with 10 microg LPS per ml blood after 10 hours of incubation. Adding PGE2-concentrations ranging from 0.03 to 10 microM to purified T cells completely abrogated T cell activation and proliferative response, which was largely reversed by adding Rp-8-Br-cAMPS. Peripheral blood mononuclear cells (PBMC) and T cells harvested from whole blood cultured in the presence of LPS ex vivo showed attenuated proliferative response (30-70%) (purified T cells and PBMC) and reduced IL-2 production (85%) upon T cell receptor (TCR)/CD3 activation with anti-CD3. The proliferation in T cells and PBMC was in part restored by Rp-8-Br-cAMPS. CONCLUSIONS: The human whole blood model of LPS-mediated T lymphocyte suppression described in this paper is time and cost efficient, as well as easy to use. PMID- 12640335 TI - Does losartan prevent cerebral edema? A preliminary study using a vascular compartment model. AB - BACKGROUND: The purpose of this study was to ascertain the effect of Losartan, a non-peptide angiotensin II receptor antagonist used for the treatment of hypertension, on the movement of proteins and fluids across the vascular compartment in ischemic cat brains. MATERIAL/METHODS: The experiments were carried out on anesthetized cats under artificial ventilation and autohemoperfusion of the brain with a stable volume of blood with the help of a resistograph. Cerebral ischemia was induced by a 15-minute arrest of the autohemoperfusion pump, tying various anastomoses in the neck region, and reducing arterial pressure to 40-30 mm Hg by hemorrhage with subsequent reinfusion of the lost blood. RESULTS: In the postischemic period in the cat brain, control experiments showed the onset of metabolic acidosis and an increase in permeability of the brain capillaries to fluids and protein molecules. Intravenous introduction of losartan, an angiotensin II receptor antagonist, at a dose of 3 mg x kg(-1) 10 minutes into the postischaemic period, enhanced the normalization of metabolic and transcapillary exchange. Thus vector permeability was reversed from blood-to-tissue in the control situation (without losartan) to tissue-to-blood during losartan administration. CONCLUSIONS: The results provide strong evidence that losartan may play a role in preventing cerebral edema, and that the renin-angiotensin system plays an important role in postischemic cerebrovascular events. PMID- 12640336 TI - The effect of brief food withdrawal on the level of free radicals and other parameters of oxidative status in the liver. AB - BACKGROUND: The aim of our study was to determine the effect of brief starvation on the level of free radicals and other parameters of oxidative status in the liver. MATERIAL/METHODS: The Wistar male rats used for the experiment were divided into two groups: Group I had free access to water and standard chow, while Group II was subjected to a 36-hour fast. The activity of antioxidant enzymes--catalase (CAT), glutathione peroxidase (GSH-Px) and Cu, Zn-superoxide dysmutase (CuZn-SOD)--was measured in the livers of both control and fasting animals. The hepatic level of free radicals and the concentration of malonyldialdehyde (MDA) were also determined in both groups of animals. RESULTS: Starvation resulted in decreased activity of CAT and CuZn-SOD, while the GSH-Px activity remained unchanged. Food deprivation also increased the level of free radicals. However, this rise was not paralleled by enhanced hepatic lipid peroxidation, measured as a change in MDA content. CONCLUSIONS: Since brief starvation causes an increase in the level of free radicals in the liver, it appears to effectively impair the tissue defence system, in part through the decreased activity of antioxidant enzymes, CAT and CuZn-SOD. PMID- 12640337 TI - Association of the ACP1 genotype with metabolic parameters upon initial diagnosis of type 1 diabetes. AB - BACKGROUND: ACP1, also called cLMWPTP (cytosolic Low Molecular Weight PTPase) is a highly polymorphic enzyme involved in the modulation of signal transduction by insulin, PDGF receptors, and T-cell receptors. The enzyme is controlled by a locus on chromosome 2, with three common codominant alleles; the corresponding six genotypes show strong variations in total enzymatic activity. The purpose of our research was to determine the relationship of ACP1 with glycemic level, ketoacidosis and HbA1C in children with Type 1 diabetes. MATERIAL/METHODS: We studied 189 consecutive children with Type 1 diabetes, from the Pediatric Clinic of Sassari University. The ACP1 genotype was determined by PCR and digestion by specific restriction enzymes. RESULTS: At initial diagnosis a strong negative correlation was observed between glycemic level and ACP1 activity, with the highest levels in genotypes with low activity. Ketoacidosis and HbA1C show a similar pattern of relationship with ACP1. A comparative analysis of the data on Type 1 diabetes with previously obtained data on Type 2 diabetes shows an opposite pattern of relationship between ACP1 and metabolic parameters. Moreover, correlation between glycemia and HbA1C in Type 1 diabetes is much weaker than in Type 2 diabetes. CONCLUSIONS: These differences suggest that the enzyme might be involved in different signal transduction pathways relevant in the pathogenesis of these two classes of diabetic disorders. It would be interesting to study the possible correlation in Type 1 diabetes between ACP1 and immunological parameters. PMID- 12640338 TI - Experience in using three different minimally invasive approaches in cardiac operations. AB - BACKGROUND: In order to reach a clear understanding of minimally invasive approaches in cardiac operations, the authors review clinical experience in using three such approaches: inferior partial median sternotomy, right anterolateral minor thoracotomy, and the right parasternal approach. MATERIAL/METHODS: Sternotomy and the three different minimally invasive approaches were applied in and 2431 and 323 patients respectively. The approaches were selected according to the circumstances of the individual case. Both external and internal cardiac structures were observed during the operations. The length of the incision, the postoperative drainage, operative time, and cardiopulmonary bypass time were investigated. The postoperative complications occurring after minimally invasive approaches were observed. RESULTS: In inferior partial median sternotomy, all structures except for the ascending aorta could be exposed well. In right anterolateral minor thoracotomy, only the structures on the right side of the heart could be exposed, but the mitral valve could also be exposed well. The exposure of the right parasternal approach was similar to that of right anterolateral minor thoracotomy. There were statistically significant differences between sternotomy and the minimally invasive approaches in terms of incision length and postoperative drainage, but no difference in operative time and cardiopulmonary bypass time. The postoperative complications of MIAs included air embolism (n = 3), chest pain (n = 9), chest wall malacia (n = 1), rib fracture (n = 2), and sternum fracture (n = 2). The total incidence of complications in minimally invasive approaches was 5.3%. CONCLUSIONS: The minimally invasive approaches can have satisfactory clinical results if the approaches are correctly chosen and performed. PMID- 12640340 TI - Laypersons' expectation of the sequelae of whiplash injury: a cross-cultural comparative study between Canada and Greece. AB - BACKGROUND: The objective of the present study is to compare the frequency and nature of expected 'whiplash' symptoms in Greece [a country where the chronic whiplash syndrome is rare or unknown] with that in Canada. MATERIAL/METHODS: A symptom checklist was administered to 2 subject groups selected from local companies in Patras Greece, and Edmonton, Canada, respectively. Subjects were asked to imagine having suffered a neck sprain [whiplash injury] with no loss of consciousness in a motor vehicle collision, and to check which, of a variety of symptoms, they would expect might arise from the injury. For symptoms they anticipated, they were asked to select the period of time they expected those symptoms to persist. RESULTS: In both the Greek and Edmontonian groups, the pattern of symptoms anticipated closely resembled the acute symptoms commonly reported by accident victims with acute neck sprain, but while up to 50% of Edmontonians also anticipated symptoms to last months or years, very few Greek subjects selected any symptoms as likely to persist. CONCLUSIONS: In Greece, despite the documented occurrence of neck sprain symptoms in individuals following motor vehicle collisions, there is a very low rate of expectation of any sequelae from this injury. What current or previous aspects of society that underlie this remain uncertain. This lack of expectation of chronicity in Greece may, in part, determine the low prevalence of the chronic whiplash syndrome there. Further studies of symptom expectation as an etiologic factor in the chronic whiplash syndrome are needed. PMID- 12640339 TI - Effects of hypolipidemic treatment on serum markers of vascular inflammation in dyslipidemic men. AB - BACKGROUND: The purpose of our study was to assess the effect of hypolipidemiant drugs on serum markers of vascular inflammation (E-Selectin, VCAM-1 and MCP-1) in dyslipidemic men without cardiovascular disease. MATERIAL/METHODS: 84 dyslipidemic men were consecutively recruited from the Lipid Unit of a tertiary hospital. The patients were placed on statins (n=44) or fibrates (n=22), depending on the lipid profile, for 4 months. In the control group (n=18), a hypolipidemiant diet alone was indicated. RESULTS: Baseline levels of VCAM-1 and MCP-1 were not correlated with the lipid profile. By contrast, baseline E Selectin levels correlated directly with glucose and triglyceride levels, and negatively with HDL-C. In multiple regression analysis, HDL-C and glucose concentrations independently influenced E-selectin levels. After treatment, we observed a significant decrease of E-Selectin levels in patients treated with statins, and the changes in E-Selectin levels were inversely associated with HDL C variations. We did not observe any changes in VCAM-1 levels after the treatment regime we used. Regarding MCP-1, a significant increase was detected in the patients receiving fibrates. In addition, the percentage increment of MCP-1 was higher in patients treated with gemfibrozil than in patients who received bezafibrate. CONCLUSIONS: We observed a reduction in E-Selectin levels after statin therapy. This finding was associated with increased HDL-C. Fibrates, especially gemfibrozil, increased MCP-1 concentrations. This deleterious effect was unrelated to changes in lipid profile, and may help explain why fibrates have less impact than statins in reducing cardiovascular disease. PMID- 12640341 TI - Baroreflex function in vasodepressive syncope: detection of early impairment. AB - BACKGROUND: Defective baroreflex function has been suggested as a potential mechanism accounting for the development of syncopal episodes. The present study was therefore aimed at assessing the non-invasive, indirect hemodynamic profile and baroreflex function by means of tilting, which is a natural stimulation crucial to physiological baroreflex activity, in syncopal patients and healthy controls. MATERIAL/METHODS: Seventeen consecutive patients with a positive response to head-up tilting and fourteen healthy subjects as controls underwent continuous and non-invasive beat-to-beat heart rate and arterial pressure measurements in order to evaluate systolic, diastolic, and dicrotic pressures, as well as heart rate. Baroreflex function was calculated as the slope of the linear regression line relating systolic arterial pressure to RR interval changes during upward and downward phases of tilting, respectively. RESULTS: When compared to healthy subjects, vasodepressive patients showed a significantly weaker correlation between systolic pressure and RR interval changes both in upward tilting, (r = 0.68 vs r = 0.91, p<0.05) and downward tilting (r = 0.48 vs r = 0.93, p<0.01). CONCLUSIONS: Our results show that an impairment in baroreflex mediated adjustment to postural challenge can be detected in syncopal patients also during upward tilting, that is, in the early phase of the test. Moreover, our investigation emphases the utility of a noninvasive, complete hemodynamic evaluation of the early phase of tilting in order to detect peculiar behaviours of pulse wave contour and related parameters. PMID- 12640342 TI - Most ventricular depolarization and repolarization abnormalities are not predictors of arrhythmic events in postinfarction patients with normal intraventricular conduction. AB - BACKGROUND: Survivors of acute myocardial infarction (AMI) are at increased risk for arrhythmic events (AE), which include sudden death (SD) or sustained ventricular tachycardia (sVT). In the prethrombolytic era, abnormal parameters of depolarization and repolarization were considered to be markers of susceptibility to these events. The purpose of the present study was to assess whether these variables should still be considered predictors of AE in postinfarction patients with normal intraventricular conduction. MATERIAL/METHODS: The study population consisted of 236 survivors of AMI, in whom the parameters of depolarization on SAECG (QRSd, LAS, RMS, LPs) and repolarization on ECG (QTc-max, QTd) were assessed before hospital discharge. The patients were followed for 18 months, and all episodes of SD and sVT were recorded. RESULTS: During long-term observation, 3 patients died from SD, and sVT occurred in 1 patient. Univariate Cox regression analysis showed that among the SA-ECG and ECG variables, only QRSd was significantly related to the incidence of AE (p<0.04). A comparison of event-free survival curves by the Kaplan-Meier method at the dichotomy limit of 119ms showed significantly worse prognosis in patients with QRSd> or =119ms (p<0.01). CONCLUSIONS: In postinfarction patients with normal intraventricular conduction treated in the acute and chronic phase of MI according to recent management strategies, QRSd> or =119ms on SAECG is associated with arrhythmic events. Other parameters of depolarization, such as LAS, RMS, and positive LPs, and also repolarization, such as prolonged QTc-max and QTd, seem to be of little use in predicting these events. PMID- 12640343 TI - Atrial fibrillation after conventional and off-pump coronary artery bypass grafting: two opposite trends in timing of atrial fibrillation occurrence? AB - BACKGROUND: Atrial fibrillation (AF) is one of the most common complications after cardiac surgery. It occurs most often between the 2nd and 3rd postoperative day. The aim of this study was to compare the daily distribution of AF incidence between patients operated on with the standard coronary artery bypass grafting (CABG) technique and off-pump coronary artery bypass grafting on the beating heart (OPCABG) without extracorporeal circulation. MATERIAL/METHODS: This was a prospective analysis of 1067 patients with coronary artery disease (CAD) operated on at our institute. 891 patients underwent CABG and 176 subjects were operated on by beating heart surgery using the OPCABG technique. Heart rhythm was analyzed continuously, and additionally a standard 12-lead ECG was performed once a day and continually in the case of clinical manifestations of arrhythmia. The observation period included 7 postoperative days (POD). RESULTS: 164 patients in the CABG group (18.43%) developed AF compared with 28 (16%) in the OPCABG group (OR=1.18; 95% CI (0.75-1.85); p=0.451). The peak of AF incidence in the CABG group appeared on the 2nd POD, while in contrast the peak of AF incidence after OPCABG was observed on the 1st POD. CONCLUSIONS: We found no statistically significant difference in AF incidence after CABG and OPCABG. Different trends were noticed in the timing of occurrence of AF in these two groups. PMID- 12640344 TI - Validation of PAH genotype-based predictions of metabolic phenylalanine hydroxylase deficiency phenotype: investigation of PKU/MHP patients from Lithuania. AB - BACKGROUND: Phenylalanine hydroxylase (PheOH) deficiency is inherited as an autosomal recessive trait. The associated hyperphenylalaninemia phenotype is highly variable, primarily due to great allelic heterogeneity in the PAH locus. The goal of our study was to assess the relationship between individual PAH locus mutations and biochemical and metabolic phenotypes in phenylketonuria (PKU) and mild hyperphenylalaninemia (MHP) patients. MATERIAL/METHODS: In this study, a total of 184 independent PAH chromosomes (92 unrelated patients with PKU and MHP residing in Lithuania) were investigated. All 13 exons of the PAH gene of all PKU probands tested were scanned for DNA sequence alterations by denaturing gradient gel electrophoresis (DGGE); mutations were identified by direct fluorescent automated sequencing or by restriction enzyme digestion analysis of the relevant exons. PAH genotype-based prediction of metabolic PhOH deficiency phenotype in PKU/MHP patients form Lithuania was estimated by the assigned value (AV) and functional hemizygosity methods. RESULTS: Our data provide evidence that a simple genotype-phenotype correlation does exist in most patients with PheOH deficiency: we observed a perfect match between the expected and observed phenotypes in 96% of the cases investigated. CONCLUSIONS: The results obtained confirm that methods of functional hemizygosity and AV sum are applicable for the estimation of the genotype-phenotype correlation in the investigated group of PKU/MHP patients. PMID- 12640345 TI - Is there any relationship between lipids and vitamin B levels in persons with elevated risk of atherosclerosis? AB - BACKGROUND: There is increasing evidence that plasma homocysteine level is an independent risk factor for atherosclerosis. Low levels of serum folates, cobalamin and pyridoxine are associated with increased risk of cardiovascular disease. Most dietary products contain cholesterol as well as methionine, so hyperlipidemia could be associated with a higher level of homocysteine and inversely with lower levels of B vitamins. The aim of this study was to investigate the differences in levels of lipids and vitamins affecting homocysteine metabolism in different groups of patients. MATERIAL/METHODS: We examined 38 healthy persons, 55 patients hospitalised for cardiac surgery, and 62 patients without clinical evidence of atherosclerosis but with one of the atherosclerosis risk factors (hypercholesterolemia, NIDDM or chronic renal insufficiency). The levels of total cholesterol, triglycerides, vitamin B12, folic acid and vitamin B6 index in serum were determined using routine laboratory methods. RESULTS: We found no association between lipids and B vitamins in any examined group. There were significant differences between concentrations of analysed parameters in all groups of patients as compared to controls. CONCLUSIONS: The lack of correlation between the levels of lipid parameters and B vitamins in serum indicates that these may be independent, additional risk factors for atherosclerosis. Higher vitamin B6 deficiency in dialysis patients is probably caused by low intake combined with the increased requirements of uremic patients. Permanent monitoring of B vitamins in serum is necessary in patients with elevated risk of atherosclerosis, as well as long-term education, careful diet planning and supplementation. PMID- 12640346 TI - Androgen and SHBG serum concentrations in late post-menopause women. AB - BACKGROUND: The purpose of our study was to evaluate androgen and SHBG concentration in the blood serum of late post-menopausal women. MATERIAL/METHODS: We examined women between 65 and 75 years of age and compared their results to those of a group of women in reproductive age. All the subjects, in addition to medical and gynecologic examinations, had determinations done of bound testosterone, free testosterone, androstenedione, dehydroepiandrosterone (DHEA) and dehydroepiandrosterone sulphate (DHEAS), sex hormone binding globulin (SHBG), and estradiol. RESULTS: The mean concentration of bound testosterone was found to be higher in postmenopausal women than in young women. The concentration of free testosterone did not vary, while the concentration of androstenedione was lower in the postmenopausal women, as were the DHEA and DHEAS concentrations. The mean SHBG concentration was higher in the postmenopausal women, and lower in obese postmenopausal women than in non-obese women. No differences were found in terms of the examined parameters between subjects with cardiovascular diseases and those without, or between smokers and non-smokers. CONCLUSIONS: The extinction of ovary function and aging of a woman is related to secondary changes of androgen concentrations in blood serum. In late postmenopausal women, the SHBG concentration increases, which is related to the increase of total testosterone concentration in blood serum. Obesity affects the reduction of SHBG concentration in blood serum in postmenopausal women. Cardiovascular diseases and smoking have no significant effect on androgen concentration and SHBG changes in blood serum in postmenopausal women. PMID- 12640347 TI - Right atrial extension of Wilms' tumor. AB - BACKGROUND: Right atrial involvement in children with Wilms' tumor (nephroblastoma) is a very rare phenomenon. CASE REPORT: The authors present four children with nephroblastoma, in whom the tumor involved the inferior vena cava and the right atrium. In two children the intracardiac tumor extension was asymptomatic, while the other two had Budd-Chiari syndrome. Therapeutic management included initial three-drug chemotherapy in three patients, and primary tumor excision in one child, since the venous involvement had been not diagnosed. In each child, the therapeutic strategy was individualized and tailored to the course of the disease. Two patients were cured. The child with bilateral Wilms' tumor died due to disease progression. Another child died suddenly at home in the course of an intermission between consecutive cycles of successful chemotherapy. CONCLUSIONS: The extension of Wilms' tumor to the great vessels and the right atrium indirectly affects the final outcome. Preoperative chemotherapy in children with Wilms' tumor invading the inferior vena cava and the right atrium is the method of choice. The extent of surgery depends on the preliminary chemotherapy results. PMID- 12640348 TI - Cortical index and size of hand bones: segregation analysis and linkage with the 11q12-13 segment. AB - BACKGROUND: Bone geometry (BG) and size (BS) are important factors in determining bone fragility. Previous studies have suggested that more than half of BG and BS variation is genetically determined. The possible chromosomal locations of genes involved in BS and BG determination have not been explored. We evaluated the extent and mode of inheritance of the radiographic hand BS index (BSI) and the metacarpal cortical index (MCI), and tested the hypothesis of linkage between these traits and the 11q 12-13 chromosomal region. MATERIAL/METHODS: Hand radiographs and blood samples were collected from 1190 individuals belonging to 349 Chuvasha nuclear families (Russian Federation). Segregation analysis was conducted on a total sample. Transmission disequilibrium testing (TDT) and model based linkage analyses (MBLA) were performed on a sub-sample of 163 families. RESULTS: The hypothesis of a major gene effect was confirmed for both studied traits. The best-fitting models were Mendelian, with an additive type of inheritance. The inferred major gene explained 50% of the CI and 40% of the BSI variation. The TDT and MBLA results did not permit confirmation of hypotheses about linkage between hand BSI and the 11q 12-13 chromosomal region, but a possible linkage between CI and that region cannot be ruled out. CONCLUSIONS: We support the hypothesis of a major gene effect in the heritability of BSI and MCI. We provide suggestive evidence for possible linkage disequilibrium between MCI and the 11q12-13 chromosomal segment (marker D11S1983), but not for a linkage between BSI and this PMID- 12640349 TI - Urinary bile acid profile in children with inborn errors of bile acid metabolism and chronic cholestasis; screening technique using electrospray tandem mass spectrometry (ES/MS/MS). AB - BACKGROUND: It is well known that urine becomes the major route for bile acid excretion in liver diseases and thus we examined bile acid profile in urine obtained from normal children and children having chronic liver diseases using electrospray tandem mass spectrometry (ES/MS/MS). MATERIAL/METHODS: Bile acid were extracted from 5 ml of urine obtained from five healthy children or from twenty patients with various liver diseases including patients with unknown chronic liver diseases, Zellweger syndrome, peroxisomal bifunctional protein deficiency disease, tyrosinema type 1, biliary atresia, and patients with progressive familial intrahepatic cholestasis (PFIC) of undetermined type. Identification and quantification of bile acids were achieved in 5 minutes using electrospray tandem mass spectrometry (ES/MS/MS). RESULTS: Urinary bile acid excretion increased in liver diseases an average of 100 times as compared to control values. There was a specific profile for different liver disease which confirms the pathology of the disease and could be used for its diagnosis. The results also show that the ions used for the diagnosis of oxo-steroid reductase deficiency disease were present in other chronic liver diseases suggesting that these atypical bile acids may not be a result of an inborn error of bile acid metabolism. CONCLUSIONS: The urinary bile acid profile obtained in this study by ES/MS/MS can be of use for the diagnosis of certain chronic liver diseases. PMID- 12640350 TI - Neuroanatomical correlates of the post-stroke aphasias studied with cerebral blood flow SPECT scanning. AB - BACKGROUND: Researchers are not in complete agreement over the extent to which specific language functions are subserved by certain brain areas. The purpose of this article was to determine neuroanatomical correlates of aphasia following cerebrovascular accident. MATERIAL/METHODS: The participants included 50 stroke patients with a single left-hemisphere lesion and residual mild to severe aphasia. Language, assessed by the Boston Diagnostic Aphasia Examination (BDAE), was affected to various degrees by a wide range of pathologies. Single-photon emission computed tomography (SPECT) images of the brain were acquired with 740 MBq (20 mCi) of Tc-99m-labeled ECD on a triple-headed gamma camera equipped with low-energy, high-resolution collimator. Correlation between reduced cerebral perfusion and the BDAE score was analyzed. RESULTS: The most prominent perfusion abnormalities in Broca's aphasia, as determined by the laterality index, were found in the frontal lobe, and to a lesser degree, the parietal lobe and striatum, whereas the most prominent deficits in Wernicke's aphasia were found in the left temporal and parietal areas. In global aphasia, SPECT images evidenced the most extensive damage throughout the perisylvian region of the left hemisphere. CONCLUSIONS: There is need for reinterpretation of the anatomical correlation of selected aphasic syndromes, especially classic Broca's and Wernicke's aphasias. The present study highlights the integrative role of some subcortical structures in language and speech functions. The results support the usefulness of regional cerebral blood flow SPECT imaging as a diagnostic aid in the post-stroke aphasias. PMID- 12640351 TI - Prognostic significance of the immunoexpression of matrix metalloproteinase MMP2 and its inhibitor TIMP2 in laryngeal cancer. AB - BACKGROUND: Matrix metalloproteinases (MMPs) are implicated in several pathological conditions, including cancer invasion and metastases. Among the MMPs associated with the tumor cell surface, gelatinase A (MMP2) is believed to be particularly important for cancer invasion. MATERIAL/METHODS: The immunoexpression of matrix metalloproteinase MMP2 and its inhibitor TIMP2 was investigated in 40 laryngeal carcinomas, in order to estimate the prognostic significance of these factors. The tissue sections were studied morphometrically using computer image analysis. RESULTS: The mean value of MMP2 immunoexpression in squamous cell carcinoma in patients without lymph node involvement was 7.36I7.3, and the mean value of MMP2 immunostaining in patients with lymph node metastases was 23.24I14.9. The difference between groups was statistically significant (P<0.001). Statistical analysis did not reveal a significant difference in TIMP2 expression between groups with metastases (mean: 11.38I7.7) and without (mean: 11.47I9.1). The survival time of patients with tumors with higher MMP2 expression (greater than the median, 9.6) was significantly shorter (P<0.004) than those with tumors showing low MMP2 expression (<9.6). The survival analysis revealed no significant difference between the survival time of patients in groups with high (> the median, 10.6) and low (<10.6) expression of TIMP2. CONCLUSIONS: Our results suggest that MMP2 expression may be of prognostic importance in laryngeal cancer. However, imbalances in the extracellular activities of matrix metalloproteinases and tissue inhibitors of matrix metalloproteinases, linked to pathological tissue destruction, are more complex and need further investigation. PMID- 12640352 TI - Propafenone versus ibutilide for post operative atrial fibrillation following cardiac surgery: neither strategy improves outcomes compared to rate control alone (the PIPAF study). AB - BACKGROUND: It is unclear whether acute conversion of atrial fibrillation (AF) with anti-arrhythmic drugs following cardiac surgery restores and/or maintains sinus rhythm or reduces hospital length of stay (LOS). MATERIAL/METHODS: A randomized prospective pilot study was conducted in 2 teaching hospitals from 3/28/98 to 8/2/99 to study the effect of the early use of ibutilide or propafenone on the duration of AF, rhythm at discharge, and LOS. A total of 42 stable patients with new AF after surgery were randomized to oral propafenone (600mg, single dose; n=20), ibutilide (1 mg up to 2 doses if necessary; n=10), or rate control only (n=12). Agents used for rate control were left to the discretion of the primary physician but beta-blockers were encouraged. RESULTS: Pre-randomization distribution of diabetes, CHF, previous AF, and the use of beta blockers were similar in all groups. At 24 hours 0%, 65% and 34% of patients in the ibutilide (p=0.01), propafenone (p=ns), and rate control groups respectively remained in AF. Although ibutilide decreased AF duration, recurrence rates were 90%, 41%, and 58% in those groups (p=ns compared to rate control). Of the 3 patients who did not convert, all received propafenone. There was no difference in LOS or rhythm at discharge. CONCLUSIONS: Ibutilide but not propafenone decreases the duration of AF after cardiac surgery and neither appears to affect LOS or rhythm at discharge. This data suggests that post operative AF is transient and routine anti-arrhythmic therapy is not necessary for the majority of patients. PMID- 12640353 TI - Oral citicoline treatment improves visual pathway function in glaucoma. AB - BACKGROUND: Increased latency and reduced amplitude of visual evoked potentials (VEP), frequently encountered in ocular hypertension or open-angle glaucoma, suggest slowed neural conduction in the visual pathways. An improvement in VEP latency and amplitude has been reported following repeated intramuscular injections of citicoline, a neuroprotective drug. Our aim was to find whether citicoline given orally would produce a similar effect. MATERIAL/METHODS: VEP latency and amplitude were measured in 21 glaucomatous eyes prior to and after two bi-weekly courses of citicoline taken orally in a dose of 1 gram/day. The treatment courses were separated by a two-week break; post-treatment VEP measurement was performed two weeks after the end of the second treatment. RESULTS: 62% of the eyes showed a response to the treatment, with VEP latency reduced from 123.5 (3.9 SEM) ms to 111.9 (1.9 SEM) ms (P=0.0008), and VEP amplitude increased from 6.56 (1.39 SEM) to 7.88 (1.16 SEM) (P=0.04). CONCLUSIONS: Citicoline given orally improves visual evoked potentials in some glaucoma patients. PMID- 12640354 TI - A comparative study of cefaclor vs amoxicillin/clavulanate in pediatric pharyngotonsillitis. AB - BACKGROUND: Pharyngotonsillitis (PT) caused by group A beta hemolytic streptococci (GABHS) is one of the most common infections of childhood. Two antibiotic suspensions, cefaclor (CEF) and amoxicillin/clavulanate (AMC), are commonly used in Poland for the treatment of PT caused by GABHS in children. MATERIAL/METHODS: This multi-center, randomized, single-blinded study was undertaken in order to compare the efficacy and safety of CEF (20 mg/kg/d) and AMC (25 mg/kg/d) in 10 days treatment of GABHS-related PT. 100 children (mean age 6 years) were enrolled into the study. Clinical and bacteriological assessments were done on the 14-18 th, and 38-45 th days after randomization. RESULTS: No GABHS strain isolated from throat smears was resistant in vitro to both antibiotics. Both antibiotics had almost 98% effectiveness at the post therapy visit. On follow-up, significantly more relapses and recurrences were observed in the AMC-treated group than in the CEF-treated group (relapse rate 21.28% vs 15.56%, p<0.02, recurrence 10.64% vs 6.66%, p<0.002). The relapse odds ratio in the AMC group was 1.7 times greater than in the CEF group, and recurrence was 1.5 times higher. There were significantly higher rates of gastrointestinal adverse events in children treated by AMC (p<0.02). CONCLUSIONS: CEF provides a clinically and bacteriologically effective treatment for children with PT caused by GABHS, comparable to AMC but significantly safer in terms of gastrointestinal side effects. AMC shows a greater risk of relapse and recurrence than CEF. PMID- 12640355 TI - The nongenomic protective effects of estrogen on the male cardiovascular system: clinical and therapeutic implications in aging men. AB - Although the preponderance of studies investigating the effects of estrogen on vasomotor tone and function have focused on women, a number of recent studies have intriguingly shown that estrogen's rapid vasodilatory properties is also preserved in men. Unlike classical steroid transcription mediated pathways, estrogen's acute vasodilatory effect is mediated by calcium dependent cell surface estrogen receptors that stimulate constitutive endothelial nitric oxide synthase (eNOS) activity. The transient release of eNOS derived nitric oxide exerts profound physiological effects on the vasculature exerting a state of cellular inhibition (i.e. vasodilation). Thus, the partial or complete attenuation of this rapid signaling system can promote endothelial dysfunction, an early pathophysiological event in atherosclerotic development. Consequently, human males experiencing age-related declines in testosterone and aromatase derived estradiol plasma levels may lose a vital cardioprotective mechanism that preserves proper endothelial function. Therapeutic strategies to preserve basal nitric oxide levels through the maintenance of normal physiological estradiol levels may confer cardiovascular benefits to aging males. PMID- 12640356 TI - Surgery, surgical education and surgical diagnostic procedures in the digital era. AB - Computers have quickly proven to be an essential part of routine, everyday clinical work. New disciplines are being developed: telesurgery (surgery at a distance), computer- and robotic-aided surgery, image-guided surgery, medical teleconsultations, postoperative telefollow-up, etc. What are the future directions of computer-aided medicine? The digital revolution in medicine, which we are currently witnessing, is leading towards the development of new surgical treatment modalities, toward surgical simulation and improved surgical teaching. The old surgical teaching rule, 'See one, do one, teach one' promulgated by Halsted is fading into history. There are new diagnostic tools expanding the available diagnostic modalities and turning their results into a more comprehensible form. Neural networks are being successfully introduced into differential diagnostics. The capacity of human hands is being increased by telemanipulation and movement downscaling. The cost of useful applications of computers is in many cases not as high as might be expected. Analogue telephone lines are often good enough for telemedical purposes, which is particularly important for developing countries. We review the current status of computer aided surgery. It is presumed that surgery in the future will rely on computers to a much larger extent than today. All surgery will be to some extent computer aided, or even completely robotic, but it can never be performed without human professional guidance, supervision and control. PMID- 12640357 TI - Bacterial adhesins and the role of sialic acid in bacterial adhesion. PMID- 12640358 TI - Health disparities: reframing the problem. AB - Health disparities are of continuing concern to the community of public health professionals. Despite concerted efforts on a number of fronts, little progress seems to have been made towards eliminating these disparities. This is due in part to a frame of reference that focuses on race and racism. While racism plays a role, continued focus on socially constructed racial groups will not lead to solutions to the problem. Humans are biological organisms and the presence of disease indicates a maladaptation between the individual human organism and its environment. Lumping together into a 'racial' group large numbers of individuals who share little in terms of phenotype, culture, and/or behavior inhibits reaching appropriate solutions. Progress will only be made when the issue of health disparities is reframed as one of phenotype/environmental mismatch. Such a frame crosscuts current racial groups. Health disparities such as hypertension, prostate cancer, low-birth-weight (LBW) infants, infant mortality, rickets, and melanoma are affected by the interactions of degree of pigmentation, amount of exposure to ultraviolet B (UVB) radiation, and levels of serum vitamin D. Reframing the problem of health disparities from one of race and racism to one of phenotype/environmental mismatch permits a solution to an otherwise intractable problem. PMID- 12640359 TI - Infertility in Poland--present status, reasons and prognosis as a reflection of Central and Eastern Europe problems with reproduction. AB - Poland has been considered a representative country for the population of Central and Eastern Europe as it has genetically homogenous population and reveals profound similarities in economical and historical processes common for all countries of the region. According to our observations there is approximately 20% of infertile couples in Poland (lack of concentration after at least 12 month of unprotected intercourse) and 40-60% of them are infertile because of the male factor. However, studies of infertility in Poland that have been conducted so far need to be verified. The changes connected with the style of life and new tendencies observed during the tedious political changes had considerable influence on childbearing rate in Poland. Additional factors diminishing the fertility rate are: the change of the status of women in a society, raising amount of everyday stress, the high rate (17%) of unemployment and the low average income. Furthermore, in Poland, the diagnostics and treatment (assisted reproductive technology) are entirely financed from the private funds of a patient. Summing up, the problem of childlessness in Central and Eastern Europe is a complex one and very alarming therefore it is necessary to work out and to implement research, preventative and therapeutic programmes strengthened through the international cooperation of specialized centers of excellence. PMID- 12640360 TI - Why are children's hospitals so busy? PMID- 12640361 TI - Evaluating resuscitation practices on the basis of evidence: the findings at first glance may seem illogical. PMID- 12640362 TI - Surfactant metabolism in newborns-insights from imprecise measurements. PMID- 12640363 TI - Abnormal weight gain and weight management: are carbohydrates the enemy? PMID- 12640364 TI - Letting the "Gini" out of the bottle: social causation and the obesity epidemic. PMID- 12640365 TI - Can diagnostic evaluations to "rule out" endocarditis be improved? PMID- 12640366 TI - Relief for the (missed) blue baby blues? PMID- 12640367 TI - So you think your mother is always looking over your shoulder?--She may be in your shoulder! PMID- 12640368 TI - The potential of DNA microarrays for the care of children. PMID- 12640369 TI - Oxidative stress in asphyxiated term infants resuscitated with 100% oxygen. AB - OBJECTIVE: To test the hypothesis that resuscitation of asphyxiated infants with pure oxygen causes hyperoxemia and oxidative stress.Study design Asphyxiated term newborn infants (n = 106) were randomly resuscitated with room air (RAR = 51) or 100% oxygen (OxR = 55). The Apgar score, time of the first cry, and establishment of a sustained pattern of respiration were recorded. Assays performed included: blood gases; reduced glutathione (GSH) and oxidized glutathione (GSSG) in whole blood; glutathione-related enzyme activities; and superoxide dismutase activity (SOD) in erythrocytes. RESULTS: The RAR group needed less time of ventilation for resuscitation (5.3 +/- 1.5 vs 6.8 +/- 1.2 min; P <.05). Pure oxygen caused hyperoxemia (PO(2), 126.3 +/- 21.8 mm Hg) that did not occur with the use of room air (PO(2), 72.2 +/- 6.8 mm Hg). GSH was decreased and GSSG, the glutathione cycle enzymes, and SOD activities were increased in both asphyxiated groups. However, the 100% oxygen-resuscitated group showed significantly greater alterations that correlated positively with hyperoxemia. CONCLUSIONS: Asphyxia causes oxidative stress in the perinatal period, and resuscitation with 100% oxygen causes hyperoxemia and increased oxidative stress. Because there are no advantages to resuscitation with 100% oxygen, room air may be preferred under certain circumstances for the resuscitation of asphyxiated neonates. PMID- 12640370 TI - Surfactant phosphatidylcholine pool size in human neonates with congenital diaphragmatic hernia requiring ECMO. AB - OBJECTIVE: We measured surfactant phosphatidylcholine (PC) pool size and half life in human congenital diaphragmatic hernia (CDH) patients who required extracorporeal membrane oxygenation (ECMO). Study design Surfactant PC pool size and half-life were measured by endotracheal administration of deuterium-labeled dipalmitoylphosphatidylcholine in 8 neonates with CDH on ECMO (CDH-ECMO), in 7 neonates with meconium aspiration syndrome on ECMO (MAS-ECMO), and in 6 ventilated infants (NON-ECMO). RESULTS: Lung PC pool size in the CDH-ECMO group was 73 +/- 17 mg/kg (mean +/- SEM), which was not significantly different from the MAS-ECMO (50 +/- 18 mg/kg) and the NON-ECMO group (69 +/- 38 mg/kg). Surfactant PC concentration in tracheal aspirates was not different between groups (~6 mg/mL). However, the percentage of palmitic acid in surfactant PC was significantly lower in the MAS-ECMO (56.3%) and the NON-ECMO (55.8%) group compared with the CDH-ECMO (67.6%) group. Surfactant PC half-life (~24 hours) was not different between the groups. A correlation was found between the surfactant PC half-life and the duration of ECMO. CONCLUSIONS: These data show no decreased surfactant PC pool size in high risk CDH patients who require ECMO. A shorter half-life of surfactant PC, indicating a faster turnover, may result in a faster improvement of the pulmonary condition during ECMO. PMID- 12640371 TI - Effects of a low-carbohydrate diet on weight loss and cardiovascular risk factor in overweight adolescents. AB - OBJECTIVES: To compare the effects of a low-carbohydrate (LC) diet with those of a low-fat (LF) diet on weight loss and serum lipids in overweight adolescents. DESIGN: A randomized, controlled 12-week trial. SETTING: Atherosclerosis prevention referral center. METHODS: Random, nonblinded assignment of participants referred for weight management. The study group (LC) (n = 16) was instructed to consume <20 g of carbohydrate per day for 2 weeks, then <40 g/day for 10 weeks, and to eat LC foods according to hunger. The control group (LF) (n = 14) was instructed to consume <30% of energy from fat. Diet composition and weight were monitored and recorded every 2 weeks. Serum lipid profiles were obtained at the start of the study and after 12 weeks. RESULTS: The LC group lost more weight (mean, 9.9 +/- 9.3 kg vs 4.1 +/- 4.9 kg, P <.05) and had improvement in non-HDL cholesterol levels (P <.05). There was improvement in LDL cholesterol levels (P <.05) in the LF group but not in the LC group. There were no adverse effects on the lipid profiles of participants in either group. CONCLUSIONS: The LC diet appears to be an effective method for short-term weight loss in overweight adolescents and does not harm the lipid profile. PMID- 12640372 TI - Alterations in height, weight, and body mass index of newborns, children, and young adults in eastern Germany after German reunification. AB - OBJECTIVE: To examine the effects of the socioeconomic changes occurring during the German reunification on anthropometric data of newborns, children, and young adults. Study design Data were drawn from different perinatal surveys and cross sectional studies that were conducted in the newly formed German states before and after reunification (1984-2000). RESULTS: The mean birth weight of newborns increased 151 g between 1984 and 1985 (n = 51,447) and 1997 (n = 83,052), whereas length increased by 0.2 cm. Among 7- to 10-year-old school children in the federal state of Saxonia between 1984 and 1985 (n = 2359) and 1999 and 2000 (n = 11,575), an increase of height by 1 to 2 cm and weight by 1.2 to 2.6 kg was observed. Average body mass index (BMI) increased by 0.7 to 0.9 kg/m(2). Among military recruits from Eastern Germany, an increase in weight and height was observed between 1985 and 1998. BMI values rose from 21.5 to 22.6 kg/m(2), and "centimeter weight" increased from 381 to 405 g/cm. CONCLUSION: The results show that the change in socioeconomic conditions was associated with alteration in anthropometric measures within a short period. This may have been caused by changes in the nutrient supply and composition as well as reduced physical activity. PMID- 12640373 TI - Does assessment of pretest probability of disease improve the utility of echocardiography in suspected endocarditis in children? AB - OBJECTIVE: To compare the yield rate (YR) of echocardiography when evaluating children with suspected infectious endocarditis (IE) in both the actual clinical setting and in the hypothetic setting where strict clinical criteria are applied. Study design Medical records of 101 children undergoing echocardiography for suspected IE were reviewed. Echocardiograms with positive findings were identified and the actual diagnostic YR was calculated. With the use of clinical criteria proposed by von Reyn (VR), the probability of IE was retrospectively classified as (1) rejected, (2) possible, or (3) probable. Theoretic YR of echocardiography was calculated for each classification. RESULTS: The actual YR of echocardiography was 12% (12/101). The YR of echocardiography by VR class was 0% in rejected, 20% in possible, and 80% in probable cases (chi(2) = 55.1, P <.0001). Echocardiography did not change the probability of IE in any patient classified as rejected, but allowed reassignment of disease probability in a significant proportion of patients with possible or probable IE. CONCLUSIONS: The YR of echocardiography was significant when clinical probability of IE was intermediate-to-high, and low, with marginal clinical utility, when clinical probability was low. Strict pretest assessment of disease probability may lead to more effective utilization of echocardiography in this population. PMID- 12640374 TI - The use of pulse oximetry to detect congenital heart disease. AB - OBJECTIVE: To evaluate whether pulse oximetry can be used as a screening test to detect congenital heart disease (CHD) in otherwise well newborns before nursery discharge. METHODS: Births at Lakeland Regional Medical Center (LRMC) who met inclusion criteria, had a single pulse oximeter reading before discharge. A normal reading was considered to be >94%. Abnormal readings were repeated on a different extremity and if still abnormal, an echocardiography was performed. The number of children with CHD and the number of echocardiograms were compared with the previous year at LRMC. RESULTS: There were 2114 infants enrolled, 88 echocardiograms (3.8%) were performed, and 43 were abnormal (yield = 48.9%), of which 12 required management (13.6%). The control group consisted of 2851 births, producing 108 echocardiograms (3.8%), 42 of which were abnormal (38.9%), and 13 required management (12.0%). No comparisons were significant. During both years, 3 children with cyanotic CHD were born. Routine pulse oximetry failed to detect a child with total anomalous pulmonary venous return. Not a single normal echocardiogram was generated by the study. CONCLUSION: Routine pulse oximetry was nearly 100% specific for detecting cyanotic CHD; as a result there was no increase in the number of echocardiograms. Routine pulse oximetry did detect a child who might not have been detected otherwise but failed to detect one that should have been detected. PMID- 12640375 TI - Zinc protoporphyrin/heme as an indicator of iron status in NICU patients. AB - OBJECTIVE: Zinc protoporphyrin/heme ratio (ZnPP/H) has been well established as an indicator of functional iron deficiency in subjects 6 months of age to adult. The primary objective of this study was to establish normative values for ZnPP/H in NICU patients and secondarily to explore the utility of this test as an indicator of iron deficiency in neonates. Study design ZnPP/H and complete blood counts were obtained weekly on consecutive NICU patients. Gestational age, growth variables, iron supplementation, erythropoietin treatment, and blood transfusions were documented. Results are reported as mean +/- SD. A value of P <.05 was considered significant. RESULTS: ZnPP/H ratios (n = 639) were evaluated from 143 infants. During the first week of life, ZnPP/H was inversely correlated with gestational age (n = 78, P <.001, r = -0.72). Maternal diabetes, growth retardation, and exposure to chorioamnionitis were independent risk factors for high ZnPP/H. Both iron supplementation and blood transfusion decreased ZnPP/H (P <.001). Erythropoietin treatment was associated with an increase in reticulocyte count and ZnPP/H (P <.001). CONCLUSIONS: ZnPP/H is inversely correlated with gestational age, and the range in all newborn infants is higher than in adults. ZnPP/H is elevated in certain infant subpopulations, which suggests that they may require additional iron supplementation. PMID- 12640376 TI - The Maternal Lifestyle Study (MLS): effects of prenatal cocaine and/or opiate exposure on auditory brain response at one month. AB - OBJECTIVE: To study absolute and interpeak latencies of the auditory brain response in infants exposed to cocaine and/or opiates in utero.Study design The sample included 477 exposed and 554 comparison infants matched for race, sex, and gestational age. Mothers were recruited at 4 urban university-based centers; most were black, receiving public assistance, and had received adequate prenatal care. Exposure was determined by meconium assay and self-report with alcohol, marijuana, and tobacco present in both groups. At 1 month, infants were tested by masked examiners with the auditory brain response. RESULTS: Analyses were conducted for exposed and comparison groups and for level of prenatal cocaine exposure with adjustment for covariates (alcohol, marijuana, tobacco, gestational age at birth, social class, and site). Heavy prenatal cocaine exposure (>/=3 days per week, first trimester) led to an increase in the I-III, I-V, and III-V interpeak latencies and to a shorter latency to peak I. Infants with prenatal opiate exposure showed a longer latency to peak V and a longer III-V interpeak latency. CONCLUSIONS: Prenatal cocaine and/or opiate exposure affects neural transmission. Detection of these effects requires a large sample with control for gestational age, other drugs, and level of cocaine use. PMID- 12640377 TI - Growth and bone density in children with mild-moderate asthma: a cross-sectional study in children entering the Childhood Asthma Management Program (CAMP). AB - OBJECTIVES: We sought to determine whether mild-moderate persistent asthma sufficient to produce a decrease in baseline lung function is associated with an adverse effect on growth and bone mineral density (BMD) in children. METHODS: This was a cross-sectional study of 1041 children, 5 to 12 years old (32% ethnic/racial minorities and 40% female), enrolled into the Childhood Asthma Management Program (CAMP). Measures of asthma severity included: Spirometry; bronchial hyperresponsiveness; duration of asthma symptoms; and symptom-based assessment of severity. Multiple regression analyses were used to relate the asthma severity on the primary outcome variables: Height by stadiometry and BMD by dual energy radiographic absorptiometry. RESULTS: The mean +/- SD height percentile was 56.0 +/- 28.5 percentile for the population. The only significant relationship between asthma severity and height percentile was with methacholine bronchoprovocation in girls (beta 2.98, P =.019, covariate multiple regression). The mean +/- SD BMD was 0.65 +/- 0.10 g/cm(2) for the population. The past use of corticosteroids did not adversely affect either growth or BMD. CONCLUSIONS: We found that mild-moderate asthma of as long as 4 to 7 years duration in children does not produce an adverse effect on linear growth or BMD. PMID- 12640378 TI - Natural killer cell dysfunction in patients with systemic-onset juvenile rheumatoid arthritis and macrophage activation syndrome. AB - OBJECTIVES: To assess natural killer (NK) and cytotoxic functions in patients with systemic-onset juvenile rheumatoid arthrithis (soJRA) complicated by macrophage activation syndrome (MAS). METHODS: NK cells (CD56+/TCRalphabeta-), NK T cells (CD56+/TCRalphabeta+) and CD8+ cells were assessed for perforin expression by flow cytometry. NK cytotoxic activity was measured after coincubation of mononuclear cells with an NK-sensitive K562 cell line. RESULTS: Two major patterns of immunologic abnormalities were detected. Four of 7 patients had decreased NK activity, low NK cell numbers, and mildly increased levels of perforin expression in CD8+ and CD56+ cytotoxic cells. Three remaining patients with MAS, however, had decreased NK activity associated with low levels of perforin expression in all cytotoxic cell populations, a pattern indistinguishable from that in carriers of perforin-deficient familial hemophagocytic lymphohistiocytosis. Remarkably, two of these patients had previous episodes of MAS. CONCLUSIONS: NK dysfunction is an immunologic abnormality common to both familial hemophagocytic lymphohistiocytosis and MAS of soJRA. The extent of NK cell abnormalities in soJRA needs to be further investigated. PMID- 12640379 TI - Vitamin A supplementation and childhood morbidity from diarrhea and respiratory infections: a meta-analysis. AB - OBJECTIVES: To perform an updated meta-analysis of the effect of vitamin A supplementation on childhood morbidity from respiratory tract infections and diarrhea. Study design A comprehensive search of the 1966 to 2000 MEDLINE database and review of the reference lists of relevant articles identified 9 randomized controlled trials dealing with morbidity from respiratory infections and diarrhea in children 6 months to 7 years of age and provided "intention-to treat" data. RESULTS: The combined results indicated that vitamin A supplementation has no consistent overall protective effect on the incidence of diarrhea (relative risk, 1.00; 95% CI, 0.94-1.07) and that it slightly increases the incidence of respiratory tract infections (relative risk, 1.08; 95% CI, 1.05 1.11). CONCLUSIONS: High-dose vitamin A supplements are not recommended on a routine basis for all preschool children and should be offered only to individuals or populations with vitamin A deficiency. PMID- 12640380 TI - Long-term outcome in children with Guillain-Barre syndrome. AB - OBJECTIVE: To determine the long-term sequelae and early predictors of sequelae for children with Guillain-Barre syndrome (GBS) after the introduction of intravenous gammaglobulin (IVIG). Study design A cross-sectional case series of children with GBS, at least 2 years after recovery, was performed. Manual muscle strength testing was done on 34 muscle groups. The functional independence measure was used to assess function. A chart review was completed, identifying acute factors that may predict long-term sequelae. RESULTS: Forty-seven children with GBS were identified, of whom 30 received IVIG. Persisting long-term muscle weakness with at least one muscle group at a grade 6 level (muscle moves the joint against gravity but only minimal resistance to an applied force) was found in 23% of cases. Each patient had a perfect score on the functional independence measure. Long-term muscle weakness was predicted by young age (P =.03) and a rapid progression to maximal weakness (P =.03). CONCLUSIONS: Despite the introduction of IVIG, 23% of children with GBS had evidence of long-term mild muscle weakness, with minimal impact on function. Young age and a rapid progression during the acute GBS period predicted long-term sequelae. PMID- 12640381 TI - Severe deficiency of the specific von Willebrand factor-cleaving protease (ADAMTS 13) activity in a subgroup of children with atypical hemolytic uremic syndrome. AB - OBJECTIVE: The von Willebrand factor-cleaving protease (VWF-cp) activity has been reported to be deficient in adults with thrombotic thrombocytopenic purpura (TTP) and generally normal in adults with hemolytic uremic syndrome (HUS). The goal of this study was to determine VWF-cp activity in children with typical postdiarrheal (d+) HUS or atypical non-postdiarrheal (d-) HUS. Study design We measured VWF-cp activity in the plasma of 64 children with either (d+) HUS (n = 41) or (d-) HUS (n = 23). RESULTS: In the acute phase of HUS, VWF-cp activity was normal (>50%) in 54 children and undetectable (<5%) in one (d+) HUS and in 6 (d-) HUS children. After a 3-month remission, the (d+) HUS patient recovered a 100% VWF-cp activity, and the 6 (d-) HUS patients kept an undetectable level. In these 6 (d-) HUS patients, the disease was characterized by a neonatal onset and several relapses (hemolytic anemia, thrombocytopenia, transient acute renal failure, cerebral ischemia), and sometimes the development of arterial hypertension or end stage renal failure. CONCLUSION: A subgroup of pediatric patients with atypical (d-) HUS, with hematologic symptoms starting at birth and a recurrent course progressively involving kidney and brain, is related to VWF-cp deficiency and actually corresponds to Upshaw-Schulman syndrome revisited as congenital TTP. PMID- 12640382 TI - Transient neonatal hyperkalemia in the antenatal (ROMK defective) Bartter syndrome. AB - OBJECTIVE: Identification of neonatal hyperkalemia as a complication of Bartter syndrome (BS), a disorder usually characterized by hypokalemic metabolic alkalosis. Study design Case-series description of a group of 12 infants with mutations in the renal potassium channel ROMK, causing one of the antenatal variants of BS. RESULTS: Prematurity, postnatal polyuria, and dehydration were seen in all cases. Plasma potassium was as high as 9.0 +/- 1.2 mmol/L and sodium as low as 124 +/- 3.5 mmol/L, appearing usually at day 3 of life and normalizing by the end of the first postnatal week. No hyperkalemia was found in 12 neonates with the variant of BS and deafness. The mean plasma potassium level during the first week of life among a group of very low-birth-weight infants with similar relative azotemia was 4.9 +/- 1 mmol/L (P <.001). The postneonatal period in the ROMK-defective children with BS was characterized by failure to thrive, hypercalciuria, nephrocalcinosis, and minimal-to-no hypokalemia. CONCLUSIONS: Early postnatal hyperkalemia, sometimes severe, may complicate antenatal BS associated with ROMK mutations. Its association with hyponatremia and hyperreninemic hyperaldosteronism may erroneously suggest the diagnosis of pseudohypoaldosteronism type 1. The expression of ROMK in both the thick ascending limb and cortical collecting duct may explain this apparently tubular maturation phenomenon. PMID- 12640383 TI - Dental abnormalities in patients with familial hypophosphatemic vitamin D resistant rickets: prevention by early treatment with 1-hydroxyvitamin D. AB - OBJECTIVE: To evaluate the dental effects of 1-hydroxylated vitamin D3 treatment in patients with familial hypophosphatemic vitamin D-resistant rickets. Study design Forty-eight children and adult patients were included in the study; 16 had received no treatment or phosphate supplements with vitamin D/25-(OH) D3 before puberty. The 32 younger ones had received phosphate supplements with 1alpha (OH)D3 from infancy. All patients were clinically examined, and panoramic and periapical radiographs were made. Evaluations of decayed, missing, or filled teeth and decayed or filled teeth indexes and of pulp ratios allowed comparison with healthy age-matched control patients. RESULTS: Poor dental health and characteristic dental anomalies were found in the 16 older patients. In contrast, the 32 younger patients had a normal dental status as regards reference ranges in healthy age-matched populations, although they still showed prominent pulp horns on deciduous teeth and increased pulp area/tooth area ratios. CONCLUSIONS: This investigation shows the beneficial effects of 1alpha-(OH)D3 treatment on the dental status of vitamin D-resistant rickets patients and emphasizes the necessity of early treatment. Remaining defects may result from early exposure of odontoblasts and surrounding osteoblasts to hypophosphatemia, before the commencement of treatment, and/or from intrinsic cell disturbances linked to the genetic alteration(s). PMID- 12640384 TI - Indication for genetic testing: a checklist for Rett syndrome. AB - OBJECTIVE: We reevaluated 49 girls with either Rett syndrome (RTT) or features of RTT who had negative test results for mutations in the MECP2 gene and compared them with 49 girls who had positive test results. The girls with MECP2-positive results included 2 girls with forme fruste and 2 with congenital RTT. Study design Based on the original diagnostic criteria for RTT, we developed a 10-item checklist with a score ranging from 0 to 12. RESULTS: If only girls with a score of 8 or more had been tested, 46% of the girls without mutations would have been excluded from testing without missing a single girl with MECP2-positive results. CONCLUSIONS: This checklist provides a simple aid for deciding whether or not a genetic test for RTT should be performed with only a minimal risk of missing girls with MECP2-positive results. PMID- 12640385 TI - Molecular analysis in Turner syndrome. AB - OBJECTIVE: The frequency of Y-chromosome material is high in Turner syndrome (TS), but the ocurrence of gonadoblastoma seems to be low. We performed a study to evaluate whether DNA analysis might be a useful tool in the evaluation of patients with TS. SUBJECTS: Unrelated patients with TS (n = 52) of Venezuelan mestizo ethnic origin were diagnosed by cytogenetic analysis as having TS. METHODS: Clinical assessment, karyotyping, endocrine evaluation, fluorescence in situ hybridization, and polymerase reaction chain analysis of the Y-chromosome loci. RESULTS: We found that 7.69% (4 of 52) patients with TS had Y-chromosome material. A low occurrence of gonadoblastoma was also found (2 of 52 [3.85%]). Two patients showed a 45,X/46,XY karyotype, and gonadoblastoma in the gonadal biopsy specimen was not found. Two patients had no Y chromosome on initial karyotype; they were positive on lymphocyte DNA to Y-sequences specific. Both patients (45,X) had bilateral gonadoblastoma. The four patients with Y-chromosome material in peripheral blood lymphocytes had Y-chromosome sequences on gonadal DNA. Fluorescence in situ hybridization confirmed their Y-chromosome origin. CONCLUSIONS: Our results suggest that the detection of Y-chromosome material should be carried out in all patients with TS and not be limited to patients with TS with cytogenetically identifiable Y chromosome and/or virilization. PMID- 12640386 TI - Turner syndrome with complex mosaic monosomy and structural aorta anomalies. PMID- 12640387 TI - An 8-year old boy with recurrent macroscopic hematuria, weight loss, and kidney failure. PMID- 12640388 TI - DNA diagnosis confirms hemoglobin deletion in newborn screen follow-up. AB - Molecular genetic confirmatory testing with polymerase chain reaction amplification is integral to neonatal hemoglobinopathy screening programs. In this study, we demonstrate applicability of polymerase chain reaction-based testing for the common deletions in blacks responsible for hereditary persistence of fetal hemoglobin. This approach will provide rapid diagnostic clarification in newborn screening follow-up. PMID- 12640389 TI - Arginase deficiency with lethal neonatal expression: evidence for the glutamine hypothesis of cerebral edema. AB - We describe a rare and lethal case of arginase deficiency in a 2-day-old female infant with encephalopathy and cerebral edema. The levels of glutamine and arginine but not ammonia were markedly elevated, lending support to the "glutamine hypothesis" as the mechanism of cerebral edema in urea cycle defects. PMID- 12640390 TI - Metabolic disease and sudden unexpected death. PMID- 12640392 TI - Growth hormone deficiency caused by pituitary stalk interruption in Diamond Blackfan anemia. PMID- 12640393 TI - Time for a fresh look at ROP screening. PMID- 12640394 TI - Verteporfin photodynamic therapy in the UK: implications of the NICE appraisal. PMID- 12640396 TI - Excision and cryosurgery of conjunctival malignant epithelial tumours. PMID- 12640395 TI - Regarding assessing G-Probe function. PMID- 12640397 TI - Safety of local anaesthesia for cataract surgery: why we should look again. PMID- 12640399 TI - Corneal sensitivity and ocular surface changes following preserved amniotic membrane transplantation for nonhealing corneal ulcers. PMID- 12640398 TI - Cataract surgery for natural rubber latex allergic patients. AB - The prevalence of reactions against natural rubber latex (NRL) is thought to be increasing in both the general public and healthcare workers. These can vary from mild benign skin reactions to bronchospasm, anaphylactic shock, and death. Difficulties exist for ophthalmic departments wishing to establish protocols in providing 'latex-free environments' for patients undergoing cataract surgery. Currently no legislation exists regarding the labelling of NRL-containing products in the United Kingdom with information on a product's NRL content provided by the manufacturer on a voluntary basis only. It is hoped this review article will act as a basic guide in the management of NRL-sensitive patients undergoing cataract surgery in the United Kingdom. PMID- 12640400 TI - Temporary amniotic membrane patching for acute chemical burns. AB - PURPOSE: To describe the surgical technique, and its usefulness, of temporary amniotic membrane patching (AMP) in the acute phase of ocular chemical injury. METHODS: Temporary AMP with modification in suture placement was performed on five eyes of five consecutive patients inflicted with acute chemical injury having a greater than grade II injury by the Roper-Hall classification. RESULTS: All patients reported herein presented with a large epithelial defect on the cornea and conjunctiva. Case 3 was classified as grade III while the other four cases were classified as grade II. The causative chemical agents were anhydrous acetic acid in Case 1, calcium oxide in Case 2, sodium hydroxide in Case 3, sodium silicate in Case 4, and sulphuric acid in Case 5. All cases experienced rapid relief of pain after AMP. Epithelialization of the cornea with improvement of visual acuity was observed in all cases when the amniotic membrane was removed within 2 weeks after surgery. During the mean follow-up of 19.6 months, the ocular surface remained stable and no cicatricial complications were noted. CONCLUSIONS: These results suggest that immediate AMP is quite useful for managing moderately severe acute ocular chemical injury by facilitating rapid epithelialization and pain relief, and securing ocular surface integrity. PMID- 12640401 TI - Multifocal ERG recording with simultaneous fundus monitoring using a confocal scanning laser ophthalmoscope. AB - PURPOSE: To assess the general feasibility of recording multifocal electroretinograms (mfERGs) with simultaneous fundus monitoring in a clinical setting. METHODS: An mfERG system (RETIscan) and a confocal scanning laser ophthalmoscope (Heidelberg Retina Angiograph) were connected to record mfERGs elicited by a pseudorandom binary m-sequence stimulus generated by a 514 nm laser source. Recordings were compared to a conventional mfERG system using monitor stimulation. A total of five subjects (two normal subjects and three patients) were examined. RESULTS: In normal volunteers records obtained by the fundus monitored mfERG showed highest response densities in the central fundus area and a decrease of response amplitudes towards the periphery paralleling cone receptor density. However, compared to the monitor stimulation, the drop-off of amplitudes as a function of eccentricity was not as clearly defined. The responses obtained from patients with retinal diseases were diminished in the areas of retinal dysfunction. CONCLUSIONS: These preliminary findings indicate that the technique of laser stimulation in principle allows for topographic retinal recording. This method might be useful, for example if the retinal position of the stimulus array is not centred onto the fovea but deviates due to fixation problems. However, further improvement of the technique appears necessary before considering routine clinical application. PMID- 12640402 TI - Assessing the 'cyclodiode G-probe' using a grey scale test: reproducibility and differences between probes. AB - AIMS: 1. To determine the number of clinicians performing cyclodiode therapy who reuse the 'G-probe' used for the delivery of cyclodiode therapy. 2. To show a simple method to assess the output of the 'G-probe' that can be used in the clinical setting. METHODS: A total of 71 questionnaires were sent to ophthalmologists who have an Oculight SLx Iris Medical Diode Laser. Ophthalmologists were asked as to whether they performed cycloablative therapy using the 'G-probe' and whether they reused the G-probe. They were also asked as to the frequency of any reuse of probes. To determine the output of the 'G probe', paper copies of a custom-made grey scale chart containing graded blocks of increasing shades of grey densities were produced. A special probe holder was made so that the G-probe tip could be held at a fixed distance from the grey scale chart. Laser burns were made on the grey scale using this arrangement and measurements of the burn size were made. After using 'standard settings' of 2000 ms and 2000 mW, 'threshold' burns were defined. Five new probes (with two different operators) were tested to assess the interprobe, interoperator, and intersheet variability of test. Probes were then tested for the burn size produced between 1000 and 3000 mW, and 1000 and 3000 ms. RESULTS: Results from the questionnaire showed that of the 44 respondents (62.0% response), 93.2% performed cyclodiode therapy with 58.5% reusing the G-probe. Among them, 56.1% reused probes on more than one occasion. Results from testing a new G-probe on the grey scale chart showed that with 'standard settings', highly reproducible burns at grey density 8 could be produced. No significant interprobe, interoperator, and intersheet variations were noted. Above 3 J of laser energy, the test could detect a 20% increase in energy settings and it was found that at levels of 4 J or above, alterations to the power setting had a greater influence on burn production than alterations to the time setting. CONCLUSIONS: This study demonstrates 1. that many clinicians in the UK reuse G-probes, 2. a simple, quick, and highly reproducible method to assess the laser output from the G-probe used for cyclodiode therapy. The method can help the ophthalmic surgeon to test the G-probe prior to commencement of therapy and with a standard treatment protocol, may produce a more predictable intraocular pressure reduction. PMID- 12640403 TI - Retinal light sensitivity in haemodialysis patients. AB - PURPOSE: The aim of the present study was to compare retinal light sensitivity between normal healthy subjects and chronic renal failure patients treated with maintenance haemodialysis (HD), as well as to determine whether there is a correlation between visual field loss and the age of HD patients, duration of HD treatment and hypertensive retinopathy in HD patients. METHODS: A total of 50 eyes of 25 HD patients (16 male, nine female) and 30 eyes of 15 controls underwent visual field testing on the C 30-2 program of the Humphrey field analyser. RESULTS: Significant reduction in retinal light sensitivity with mean deviation (MD) P values less than 5% was found in 36% (18 eyes), pattern standard deviation (PSD) P values less than 5% in 16 eyes (32%) and corrected pattern standard deviation (CPSD) P values less than 5% in 16 eyes (32%) of HD patients. In control group, all MD, PSD, and CPSD P values were within normal limits. No correlation between reduction of retinal light sensitivity and age or duration of dialysis treatment was observed in HD patients. The reduction of retinal light sensitivity was significantly greater in HD patients with hypertensive retinopathy. CONCLUSION: In 36% of eyes from our HD patients without ophthalmoscopically evident arteriolar occlusion on fundus examination, a significant reduction in retinal light sensitivity was observed. The reduction was significantly greater in HD patients with hypertensive retinopathy. PMID- 12640404 TI - Retinopathy of prematurity in practice. I: screening for threshold disease. AB - AIMS: To review an 11-year period of screening for retinopathy of prematurity (ROP) in the north of England by a single ophthalmologist. To identify the gestational ages and birth weights of babies reaching different stages of ROP. To investigate the workload involved in screening to detect threshold ROP, and that the practical outcomes had narrower inclusion criteria for screening. To identify babies treated for threshold disease. METHODS: During the period August 1987 October 1998, babies were screened according to the national guidelines and the results were prospectively entered onto a computerised database. These data were then systematically reviewed. RESULTS: Data were available for 484 babies: 203 (41.9%) developed any ROP, 46 (9.5%) reached stage 3 ROP, and 25 (5.2%) reached threshold and were treated. Data on 425 babies showed them to require an average of 2.3 screening examinations per baby. It took an average of 39 screening examinations to detect one case of threshold ROP. The more premature and lighter birth weight babies required the most examinations. Therefore, restricting the inclusion criteria for screening would only have reduced the total number of screenings modestly and could have allowed us to miss two of our threshold cases who were both of 30 weeks gestational age and >1400 g birth weight. CONCLUSIONS: Screening is time consuming but worthwhile in view of the benefits of treatment. As applied to babies in the north of England, the current national screening criteria are satisfactory. The results of treatment of the babies identified in this study are presented in the accompanying paper. PMID- 12640405 TI - Retinopathy of prematurity in practice. II: long-term results following treatment for threshold disease. AB - OBJECTIVE: To determine the long-term outcome of treatment for threshold retinopathy of prematurity (ROP) in one UK unit. METHODS: A total of 35 infants diagnosed with threshold ROP were treated by a single ophthalmologist (DGC) between 1987 and 1998. The parents of the 29 surviving patients were contacted; 21 brought their child for special review of acuity, motility, refraction and fundoscopy (Group 1). Of the eight patients who were unable to attend, six patients (Group 2) had sufficient follow-up detail in their records to be included in the study. RESULTS: A total of 40 eyes in the 21 patients in Group 1 had been treated; 10 had received cryotherapy (eight bilateral) and 11 had bilateral laser. Mean follow-up was 5 years (range 2-12 years). A total of 23 treated eyes (in 16 patients) had an acuity of 6/12+ (57.5% of treated eyes). Four eyes of three patients had no light perception. Seven eyes in four patients had myopia of more than 3 D. Eight patients had a manifest squint: six esotropia, two exotropia. The retinal status of 36 eyes (90%) showed a favourable outcome. The four blind eyes had stage 5 disease. In Group 2, four eyes of two patients (33%) had 6/12+ acuity recorded; five eyes in three patients had stage 5 disease and no perception of light. In total, a favourable structural outcome was achieved in 81.4% and favourable functional outcome observed in 74.6%. CONCLUSION: These results show that the favourable outcomes reported in the CRYO ROP trial can be achieved in routine practice and maintained after long follow up. PMID- 12640406 TI - Surgically induced miosis during phacoemulsification in patients with diabetes mellitus. AB - PURPOSE: To assess the incidence of surgically induced miosis during phacoemulsification in diabetic patients. METHODS: A total of 76 patients with diabetes mellitus were compared to 76 age- and race-matched controls. A combination of cyclopentolate 1%, phenylephrine 2.5% and diclofenac sodium 0.1% was applied topically 60, 45 and 30 min before surgery. Adrenaline mixed with buffered saline solution was used for irrigation during surgery. The procedure included phacoemulsification and implantation into the bag of a foldable acrylic implant. Measurements of the horizontal pupillary diameter were taken at three stages: before corneal incision, after phacoemulsification, and at the end of surgery. The duration of phacoemulsification was also recorded. RESULTS: Surgically induced miosis or dilation of the pupil was defined as constriction or dilation noted at any interval during surgery. The pairs of diabetic-control were grouped into three groups: those in which constriction was noted, those in which dilation was noted, and those in which there was no change in pupil size during the procedure. Surgically induced miosis was noted more often in the diabetics (McNemar's test, chi(2), P=0.016). The mean pupil size at the beginning of surgery was 7.38 (+/-0.95) mm in the diabetics as compared to 7.65 (+/-0.89) mm in the control group. No statistically significant difference was noted between the two groups (paired t-test, P=0.07). The mean (+/-SD) duration of phacoemulsification in the diabetic group was 2.31 (+/-1) min as compared to 2.05 (+/-0.82) min in the control group. No statistically significant difference was found between the two groups (paired t-test, P=0.08). CONCLUSION: Surgically induced miosis occurred more often in the diabetics. Therefore, it is advisable that phacoemulsification in this group of patients is undertaken by an experienced surgeon. PMID- 12640407 TI - The efficacy and safety of 0.75% levobupivacaine vs 0.75% bupivacaine for peribulbar anaesthesia. AB - BACKGROUND: Levobupivacaine, the S(-)-enantiomer of racemic bupivacaine, is associated with a similar efficacy but a reduced potential for cardiovascular and central nervous system toxicity than racemic bupivacaine. Thus, this prospective, randomised, double-masked study was undertaken to assess the efficacy and safety of 0.75% levobupivacaine vs 0.75% bupivacaine, each with hyaluronidase, for peribulbar anaesthesia. METHODS: A total of 60 patients undergoing elective anterior segment surgery were randomly allocated to receive either agent by a single, inferotemporal peribulbar injection technique, supplemented with a medial canthus injection if necessary. Ocular akinesia and orbicularis oculi function were assessed by scoring systems at 2 min intervals until satisfactory akinesia was achieved, and movements were reassessed on the day after surgery to confirm regression of the block. RESULTS: The time taken to reach a state of satisfactory anaesthesia and akinesia was deemed to be the primary measure of efficacy. Both agents achieved this in a similar median time of 2 min after receiving 5 ml of the injectate, and the treatment difference was not statistically significant (P=0.24). Blood samples from the first 20 patients were taken at intervals up to 4 h. These were analysed for plasma levels and confirmed similar plasma concentration vs time profiles for the two agents. Seven patients in each group (23%) complained of pain on injection but the technique was generally well tolerated. Two patients in the levobupivacaine group experienced serious adverse events, but neither was considered related to the study medication. The most common minor post-operative adverse event was prolongation of the local anaesthetic block, which was reported by nine patients (four in the levobupivacaine group and five in the bupivacaine group). CONCLUSIONS: Levobupivacaine and bupivacaine are equally successful in achieving clinically satisfactory peribulbar anaesthesia with few adverse effects. PMID- 12640408 TI - Beta irradiation: new uses for an old treatment: a review. AB - Beta radiation has a long history as a treatment modality in ophthalmology. It is a convenient and practical method of applying radiation and has the advantage of minimal tissue penetration. There has been a recent resurgence in the use of beta radiation in other areas in medicine, such as the prevention of restenosis after coronary artery stenting. Beta radiation has been shown in vitro and in vivo to inhibit proliferation of human Tenon's fibroblasts, which enter a period of growth arrest but do not die. Effects on the cell cycle controller p53 have been shown to be important in this process. In ophthalmology, beta radiation has been used widely for the treatment of pterygium and is under evaluation for treatment of age-related macular degeneration and for controlling wound healing after glaucoma drainage surgery. In this latter role, beta radiation may be particularly appropriate for use in developing countries to improve the results of trabeculectomy while potentially avoiding some of the side effects of other antimetabolites. PMID- 12640409 TI - Malignant eyelid tumours in Taiwan. AB - AIMS: To describe the clinical characteristics of patients with eyelid cancers in Taiwan. METHODS: Between 1980 and 2000, 127 (58 males and 69 females) patients (mean age 62.6; range 10-91 years) with histologically confirmed eyelid cancers were retrospectively evaluated at the National Taiwan University Hospital and Far Eastern Memorial Hospital in Taiwan. Clinical data of all patients were reviewed from medical records. The mean follow-up period was 62.4 months (range 3-240) for 113 patients. RESULTS: The 127 eyelid cancers included 79 basal cell carcinomas (62.2%), 30 sebaceous gland carcinomas (23.6%), 11 squamous cell carcinomas (8.7%), five malignant melanomas (3.9%), one Kaposi's sarcoma (0.8%), and one metastatic cancer (0.8%). Tumours developed more commonly in the lower (37.0%) than the upper eyelid (33.9%). The clinical accuracy in predicting eyelid malignancy was 90.5%. Primary treatment modality was mainly surgical excision. The recurrence, metastasis, and mortality rates at 5 years were 15.2, 11.7, and 7.3%, respectively, for all eyelid malignancies. Rates of sebaceous gland carcinoma recurrence, metastasis, and mortality were significantly higher (P<0.05) than those of basal cell carcinoma. The mean interval of recurrence or metastasis after primary treatment was 26.3 months (range 4-112) for all eyelid cancers. CONCLUSIONS: Although basal cell carcinoma is the most common eyelid cancer in Taiwan, sebaceous gland carcinoma is also common. Of the two, basal cell carcinoma has a better prognosis and sebaceous gland carcinoma has a higher mortality and therefore should be treated much more aggressively. Long-term follow-up is needed after treatment of malignant eyelid tumours. PMID- 12640410 TI - Vitreous surgery in the management of chronic endogenous posterior uveitis. AB - OBJECTIVES: There is evidence that pars plana vitrectomy (PPV) has a beneficial effect on the clinical course of chronic endogenous posterior uveitis (EPU) possibly by physically removing any resident inflammatory cells with the vitreous. We assessed the anatomical and therapeutic effects of PPV performed on patients with chronic EPU for any indication. PATIENTS AND METHODS: Retrospective review of 41 eyes of 38 consecutive patients with EPU who underwent a PPV for any reason, over a 5-year period. The mean age of the patients was 36.2 years, 46% of the eyes had intermediate uveitis, 32% panuveitis, and 22% posterior uveitis. The visual acuity, disease activity, and the requirement for medications to control it were recorded for 12 months pre- and postoperatively. RESULTS: Overall, 61% of the eyes gained more than 2 Snellen lines (P<0.001) and the incidence of cystoid macular oedema (CMO) significantly reduced from 44 to 20% (P<0.05). Postoperatively, there was a significant decrease in the recurrence rate of intermediate uveitis, posterior uveitis, and panuveitis (P<0.001). The use of systemic and local depot immunosuppressive agents did not change over the study period, although the use of topical agents increased (P<0.05). CONCLUSION: PPV appears to have a beneficial effect on the clinical course of EPU in selected cases. This may be mediated by the physical clearance of inflammatory debris, the anti-inflammatory effect of replacing vitreous by aqueous humour, by a reduction of CMO and/or the anatomical correction of sight-threatening retinal pathology. PMID- 12640411 TI - Excision and cryosurgery in the treatment of conjunctival malignant epithelial tumours. AB - PURPOSE: To evaluate the long-term results of combined treatment with excision and cryosurgery for malignant epithelial tumours of the conjunctiva. METHODS: We retrospectively reviewed the medical records of patients diagnosed with conjunctival malignant epithelial tumours and treated with excision and cryosurgery between 1980 and 1998. The demographic features of the patients, characteristics of the tumours, and the peculiarities of the recurrences were recorded. The SPSS V 9.05 computed program was used for the statistical analysis of the results. Kaplan-Meier plots were used to determine the recurrence-free survival, and the logrank test was used to compare the recurrences according to histological classification. RESULTS: A total of 55 subjects (57 eyes), 19 female and 36 male, were included in the study. Their ages ranged between 15 and 82 years (mean 55+/-14 years). Histological diagnosis was conjunctival intraepithelial neoplasia (CIN) in 26 (46%) and invasive squamous cell carcinoma (SCC) of the conjunctiva in 31 eyes (54%). All eyes were treated with excision and cryosurgery with nitrous oxide probe. During the follow-up period of 6 months to 15 years, seven (12.3%) recurrences were observed. The success rate for patients with CIN and invasive SCC was 88.5 and 87.1%, respectively. There was no difference for recurrence rates between CIN and invasive SCC groups according to logrank test (P=0.68). CONCLUSIONS: Over the long-term follow-up, cryosurgery following excision is a successful method in the treatment of conjunctival intraepithelial tumours and invasive squamous cell carcinoma of conjunctiva with favourable outcome and minimal complications. PMID- 12640412 TI - Management of band keratopathy with excimer phototherapeutic keratectomy: visual, refractive, and symptomatic outcome. AB - PURPOSE: To report on the outcome of excimer phototherapeutic keratectomy (PTK) in the management of band keratopathy. METHODS: Analysis was performed on patients undergoing PTK for band keratopathy between January 1996 and December 1999. Patients were assessed as to whether the indication for PTK was for visual rehabilitation (visual group) or for ocular surface improvement (nonvisual group). Further data including pre- and postoperative best-corrected visual acuity, pre- and postoperative spherical equivalent (SE), and associated or previous ocular pathology were collected. RESULTS: A total of 45 eyes of 33 patients underwent PTK for band keratopathy. In the visual group, there were 33 eyes of 22 patients, and in the nonvisual group 12 eyes of 11 patients. The visual acuity was improved or unchanged in 18/33 eyes (55%) of the visual group, while there was loss of one or more lines of acuity in 15/33 (45%). Loss of acuity was attributed to other factors in 11/15 eyes. There was a significant myopic shift (P<0.5) from the mean preoperative SE of +1.42 D (SD+/-3.21 D) to the mean postoperative SE of +0.32 (SD+/-3.26 D). Of the eyes in the nonvisual group, 10/12 (83%) had significant improvement of symptoms. CONCLUSION: Excimer PTK is an effective procedure in the management of band keratopathy whether it is performed for visual rehabilitation or for ocular surface improvement to prevent erosions. Concurrent pathology is common and may limit the extent of visual improvement postoperatively. Factors associated with the myopic shift seen in this series may be epithelial remodelling, concurrent ocular pathology, or inaccuracy of preoperative refractive data. PMID- 12640413 TI - Visual performance of acrylic and PMMA intraocular lenses. AB - AIM: To evaluate the quality of visual functions after cataract surgery and intraocular lens (IOL) implantation with different lens materials and compare the results with age-matched subjects with clear phakic eyes. METHODS: Control and pseudophakic groups involved individuals aged between 50 and 75 years, without any accompanying ocular or systemic disease. In all, 50 eyes implanted with foldable acrylic IOLs, and 41 eyes implanted with polymethyl-methacrylate (PMMA) IOLs were compared with 45 phakic eyes as controls. Visual functions were evaluated for contrast sensitivity function and glare disability. The results were compared statistically using one-way analysis of variance (ANOVA). RESULTS: At high luminance levels, the difference among groups for contrast sensitivity was statistically significant for all spatial frequencies (P<0.05). Although the acrylic IOL-implanted eyes had better results at all three spatial frequencies, no significant difference existed between the two study groups for these spatial frequencies (P>0.05). Glare disability scores were significantly higher in the PMMA-IOL group compared to the control and acrylic-IOL groups. CONCLUSION: The visual quality achieved in pseudophakic eyes was not as good as in clear phakic eyes in regard to contrast sensitivity and glare. However, acrylic IOLs fared better than PMMA IOLs. PMID- 12640414 TI - Vitrectomy for phacolytic glaucoma in a patient with ectopia lentis et pupillae. AB - This case report highlights the successful management, by vitrectomy alone, of a case of chronic phacolytic glaucoma secondary to a dislocated hypermature lens in the vitreous of a patient with ectopia lentis et pupillae (ELP). The features and complications of ELP are discussed. PMID- 12640415 TI - New retinal detachment following removal of a scleral explant. PMID- 12640416 TI - Unilateral pellucid marginal degeneration. PMID- 12640417 TI - Miragel explant fragmentation 10 years after scleral buckling surgery. PMID- 12640418 TI - Acute visual loss as an early manifestation of metastatic neuroblastoma. PMID- 12640419 TI - Primary nonfamilial ocular amyloidosis. PMID- 12640422 TI - Preseptal cellulitis in systemic onset juvenile idiopathic arthritis. PMID- 12640420 TI - Bilateral third-nerve palsy with aberrant regeneration in Guillain-Barre syndrome. PMID- 12640421 TI - Factitious keratoconjunctivitis (not another case of ocular Munchausen's syndrome). PMID- 12640423 TI - Iatrogenic retinal tear and vitreous haemorrhage with Rycroft cannula during phacoemulsification cataract surgery. PMID- 12640424 TI - Spontaneous suprachoroidal haemorrhage following a valsalva manoeuvre. PMID- 12640425 TI - Complete spontaneous regression of a basal cell carcinoma. PMID- 12640426 TI - Delayed allergic reaction to hyaluronidase: a rare sequel to cataract surgery. PMID- 12640427 TI - Iris heterochromia after vertical squint surgery in a patient with SLE. PMID- 12640428 TI - Extrusion of a radon seed after 40 years, a case of mistaken identity. PMID- 12640429 TI - Optic disc metastasis presenting as an initial sign of recurrence of adenoid cystic carcinoma of the larynx. PMID- 12640430 TI - A delayed complication of cataract surgery in a patient with pseudoexfoliation: dislocation of the intraocular lens. PMID- 12640431 TI - Bilateral microphthalmos and orbital cyst. PMID- 12640432 TI - Should we discontinue tamoxifen in a patient with vision-threatening ocular toxicity related to low-dose tamoxifen therapy? PMID- 12640433 TI - The ocular hazards of egg throwing. PMID- 12640434 TI - Penetrating eye injury following a snake attack. PMID- 12640435 TI - Consanguinity and disease coincidence. PMID- 12640436 TI - Central retinal vein obstruction with cilio-retinal infarction. PMID- 12640438 TI - Orbital cellulitis following corneal gluing under sub-Tenon's local anaesthesia. PMID- 12640439 TI - The Royal College of Ophthalmologists Cataract Surgery Guidelines: what can patients see with their operated eye during cataract surgery? PMID- 12640440 TI - Young adults with advanced glaucoma. PMID- 12640442 TI - Potassium channel gating observed with site-directed mass tagging. AB - Potassium channels allow the selective flow of K+ ions across otherwise impermeable membranes. During a process called gating, these channels undergo a conformational change that proceeds from a closed to an open state. The closed state of KcsA, a prokaryotic potassium channel, has been structurally well characterized with equilibrium structural techniques. However, attempts to obtain a structural description of the gating transition of the channel have been hampered because the open state is only transiently occupied and, therefore, not readily accessible to such techniques. Here we describe a non-equilibrium technique that we call site-directed mass tagging and use this technique to probe the conformational change that KcsA undergoes during gating. The results indicate that KcsA is a dynamically modular molecule; the extracellular half of the membrane-spanning region is held rigid during gating, while the intracellular half undergoes a significant conformational change. PMID- 12640443 TI - A HEX-1 crystal lattice required for Woronin body function in Neurospora crassa. AB - The Woronin body is a dense-core vesicle specific to filamentous ascomycetes (Euascomycetes), where it functions to seal the septal pore in response to cellular damage. The HEX-1 protein self-assembles to form this solid core of the vesicle. Here, we solve the crystal structure of HEX-1 at 1.8 A, which provides the structural basis of its self-assembly. The structure reveals the existence of three intermolecular interfaces that promote the formation of a three-dimensional protein lattice. Consistent with these data, self-assembly is disrupted by mutations in intermolecular contact residues and expression of an assembly defective HEX-1 mutant results in the production of aberrant Woronin bodies, which possess a soluble noncrystalline core. This mutant also fails to complement a hex-1 deletion in Neurospora crassa, demonstrating that the HEX-1 protein lattice is required for Woronin body function. Although both the sequence and the tertiary structure of HEX-1 are similar to those of eukaryotic initiation factor 5A (eIF-5A), the amino acids required for HEX-1 self-assembly and peroxisomal targeting are absent in eIF-5A. Thus, we propose that a new function has evolved following duplication of an ancestral eIF-5A gene and that this may define an important step in fungal evolution. PMID- 12640444 TI - Loading direction regulates the affinity of ADP for kinesin. AB - Kinesin is an ATP-driven molecular motor that moves processively along a microtubule. Processivity has been explained as a mechanism that involves alternating single- and double-headed binding of kinesin to microtubules coupled to the ATPase cycle of the motor. The internal load imposed between the two bound heads has been proposed to be a key factor regulating the ATPase cycle in each head. Here we show that external load imposed along the direction of motility on a single kinesin molecule enhances the binding affinity of ADP for kinesin, whereas an external load imposed against the direction of motility decreases it. This coupling between loading direction and enzymatic activity is in accord with the idea that the internal load plays a key role in the unidirectional and cooperative movement of processive motors. PMID- 12640445 TI - Crystal structure of the ascorbate peroxidase-ascorbate complex. AB - Heme peroxidases catalyze the H2O2-dependent oxidation of a variety of substrates, most of which are organic. Mechanistically, these enzymes are well characterized: they share a common catalytic cycle that involves formation of a two-electron, oxidized Compound I intermediate followed by two single-electron reduction steps by substrate. The substrate specificity is more diverse--most peroxidases oxidize small organic substrates, but there are prominent exceptions- and there is a notable absence of structural information for a representative peroxidase-substrate complex. Thus, the features that control substrate specificity remain undefined. We present the structure of the complex of ascorbate peroxidase-ascorbate. The structure defines the ascorbate-binding interaction for the first time and provides new rationalization of the unusual functional features of the related cytochrome c peroxidase enzyme, which has been a benchmark for peroxidase catalysis for more than 20 years. A new mechanism for electron transfer is proposed that challenges existing views of substrate oxidation in other peroxidases. PMID- 12640446 TI - Neuropathology of human Alzheimer disease after immunization with amyloid-beta peptide: a case report. AB - Amyloid-beta peptide (Abeta) has a key role in the pathogenesis of Alzheimer disease (AD). Immunization with Abeta in a transgenic mouse model of AD reduces both age-related accumulation of Abeta in the brain and associated cognitive impairment. Here we present the first analysis of human neuropathology after immunization with Abeta (AN-1792). Comparison with unimmunized cases of AD (n = 7) revealed the following unusual features in the immunized case, despite diagnostic neuropathological features of AD: (i) there were extensive areas of neocortex with very few Abeta plaques; (ii) those areas of cortex that were devoid of Abeta plaques contained densities of tangles, neuropil threads and cerebral amyloid angiopathy (CAA) similar to unimmunized AD, but lacked plaque associated dystrophic neurites and astrocyte clusters; (iii) in some regions devoid of plaques, Abeta-immunoreactivity was associated with microglia; (iv) T lymphocyte meningoencephalitis was present; and (v) cerebral white matter showed infiltration by macrophages. Findings (i)-(iii) strongly resemble the changes seen after Abeta immunotherapy in mouse models of AD and suggest that the immune response generated against the peptide elicited clearance of Abeta plaques in this patient. The T-lymphocyte meningoencephalitis is likely to correspond to the side effect seen in some other patients who received AN-1792 (refs. 7-9). PMID- 12640447 TI - Predicting hepatitis B virus-positive metastatic hepatocellular carcinomas using gene expression profiling and supervised machine learning. AB - Hepatocellular carcinoma (HCC) is one of the most common and aggressive human malignancies. Its high mortality rate is mainly a result of intra-hepatic metastases. We analyzed the expression profiles of HCC samples without or with intra-hepatic metastases. Using a supervised machine-learning algorithm, we generated for the first time a molecular signature that can classify metastatic HCC patients and identified genes that were relevant to metastasis and patient survival. We found that the gene expression signature of primary HCCs with accompanying metastasis was very similar to that of their corresponding metastases, implying that genes favoring metastasis progression were initiated in the primary tumors. Osteopontin, which was identified as a lead gene in the signature, was over-expressed in metastatic HCC; an osteopontin-specific antibody effectively blocked HCC cell invasion in vitro and inhibited pulmonary metastasis of HCC cells in nude mice. Thus, osteopontin acts as both a diagnostic marker and a potential therapeutic target for metastatic HCC. PMID- 12640448 TI - Clonality analysis after retroviral-mediated gene transfer to CD34+ cells from the cord blood of ADA-deficient SCID neonates. AB - A clinical trial of retroviral-mediated transfer of the adenosine deaminase (ADA) gene into umbilical cord blood CD34(+) cells was started in 1993. ADA-containing peripheral blood mononuclear cells (PBMCs) have persisted in patients from this trial, with T lymphocytes showing the highest prevalence of gene marking. To gain a greater understanding of the nature and number of the transduced cells that were engrafted, we used linear amplification-mediated PCR (LAM-PCR) to identify clonal vector proviral integrants. In one patient, a single vector integrant was predominant in T lymphocytes at a stable level over most of the eight-year time span analyzed and was also detected in some myeloid samples. T-cell clones with the predominant integrant, isolated after eight years, showed multiple patterns of T-cell receptor (TCR) gene rearrangement, indicating that a single pre-thymic stem or progenitor cell served as the source of the majority of the gene-marked cells over an extended period of time. It is important to distinguish the stable pattern of monoclonal gene marking that we observed here from the progressive increase of a T-cell clone with monoclonal gene marking that results from leukemic transformation, as observed in two subjects in a clinical trial of gene therapy for X-linked severe combined immunodeficiency (SCID). PMID- 12640449 TI - Alzheimer disease's double-edged vaccine. PMID- 12640450 TI - T cell fitness determined by signal strength. AB - Two potential outcomes confront proliferating antigen-stimulated naive T cells: differentiation to effector and memory cells, or deletion. How stimulation affects cell fate is unclear. Autonomous CD8+ T cell differentiation has been proposed, but this does not explain the abortive proliferation of T cells induced by immature dendritic cells. Here we show that human and mouse CD4+ and CD8+ T cells receiving short or weak stimulation of the T cell receptor proliferate in response to interleukin 2 (IL-2) but are not 'fit' because they die by neglect, fail to proliferate in response to IL-7 and IL-15 and disappear in vivo. Conversely, prolonged or strong stimulation promotes 'fitness' by enhancing survival and cytokine responsiveness. Our results are consistent with the concept that signal strength drives progressive T cell differentiation and the acquisition of fitness. PMID- 12640451 TI - Dynamic programming of CD8+ T lymphocyte responses. AB - The initial encounter with an antigen-presenting cell (APC) is the primary force behind the expansion, differentiation and survival of naive T cells. Using an APC that permits temporal control of priming, we examined whether the duration of antigenic stimulation can influence the functional development of CD8+ cytotoxic T lymphocytes (CTLs) in vivo. Whereas CTLs given a 4-h stimulus underwent an abortive clonal expansion with transient surface CD25 expression, those given a 20-h stimulus sustained CD25 up-regulation, proliferated extensively, and efficiently mediated destruction of peripheral target tissues. Our results show that an instructional program preceding the first cell division integrates differences in signal strength into the decision to activate versus tolerize specific CTL clones. PMID- 12640452 TI - A splicing mutation affecting expression of ataxia-telangiectasia and Rad3 related protein (ATR) results in Seckel syndrome. AB - Seckel syndrome (OMIM 210600) is an autosomal recessive disorder characterized by intrauterine growth retardation, dwarfism, microcephaly and mental retardation. Clinically, Seckel syndrome shares features in common with disorders involving impaired DNA-damage responses, such as Nijmegen breakage syndrome (OMIM 251260) and LIG4 syndrome (OMIM 606593). We previously mapped a locus associated with Seckel syndrome to chromosome 3q22.1-q24 in two consanguineous Pakistani families. Further marker analysis in the families, including a recently born unaffected child with a recombination in the critical region, narrowed the region to an interval of 5 Mbp between markers D3S1316 and D3S1557 (145.29 Mbp and 150.37 Mbp). The gene encoding ataxia-telangiectasia and Rad3-related protein (ATR) maps to this region. A fibroblast cell line derived from an affected individual displays a defective DNA damage response caused by impaired ATR function. We identified a synonymous mutation in affected individuals that alters ATR splicing. The mutation confers a phenotype including marked microcephaly (head circumference 12 s.d. below the mean) and dwarfism (5 s.d. below the mean). Our analysis shows that UV-induced ATR activation can occur in non-replicating cells following processing by nucleotide excision repair. PMID- 12640453 TI - Polyalanine expansion and frameshift mutations of the paired-like homeobox gene PHOX2B in congenital central hypoventilation syndrome. AB - Congenital central hypoventilation syndrome (CCHS or Ondine's curse; OMIM 209880) is a life-threatening disorder involving an impaired ventilatory response to hypercarbia and hypoxemia. This core phenotype is associated with lower penetrance anomalies of the autonomic nervous system (ANS) including Hirschsprung disease and tumors of neural-crest derivatives such as ganglioneuromas and neuroblastomas. In mice, the development of ANS reflex circuits is dependent on the paired-like homeobox gene Phox2b. Thus, we regarded its human ortholog, PHOX2B, as a candidate gene in CCHS. We found heterozygous de novo mutations in PHOX2B in 18 of 29 individuals with CCHS. Most mutations consisted of 5-9 alanine expansions within a 20-residue polyalanine tract probably resulting from non homologous recombination. We show that PHOX2B is expressed in both the central and the peripheral ANS during human embryonic development. Our data support an essential role of PHOX2B in the normal patterning of the autonomous ventilation system and, more generally, of the ANS in humans. PMID- 12640454 TI - Loss of neuropathy target esterase in mice links organophosphate exposure to hyperactivity. AB - Neuropathy target esterase (NTE) is involved in neural development and is the target for neurodegeneration induced by selected organophosphorus pesticides and chemical warfare agents. We generated mice with disruptions in Nte, the gene encoding NTE. Nte(-/-) mice die after embryonic day 8, and Nte(+/-) mice have lower activity of Nte in the brain and higher mortality when exposed to the Nte inhibiting compound ethyl octylphosphonofluoridate (EOPF) than do wild-type mice. Nte(+/-) and wild-type mice treated with 1 mg per kg of body weight of EOPF have elevated motor activity, showing that even minor reduction of Nte activity leads to hyperactivity. These studies show that genetic or chemical reduction of Nte activity results in a neurological phenotype of hyperactivity in mammals and indicate that EOPF toxicity occurs directly through inhibition of Nte without the requirement for Nte gain of function or aging. PMID- 12640455 TI - Sir3p phosphorylation by the Slt2p pathway effects redistribution of silencing function and shortened lifespan. AB - An organism's lifespan is modulated by environmental conditions. When nutrients are abundant, the metabolism of many organisms shifts to growth or reproduction at the expense of longer lifespan, whereas a scarcity of nutrients reverses this shift. These correlations suggest that organisms respond to environmental changes by altering their metabolism to promote either reproduction and growth or long life. The only previously reported signaling mechanism involved in this response is the nutrient-responsive insulin/insulin-like growth factor-1 receptor pathway. Here we report another pathway that controls the length of yeast lifespan. Commitment to cell growth activates the Slt2p MAP kinase pathway, which phosphorylates the transcriptional silencing protein Sir3p, resulting in a shorter lifespan. Elimination of the Sir3p phosphorylation site at Ser275 extended lifespan by 38%. Lifespan extension occurs by a mechanism that is independent of suppressing rDNA recombination. Thus, Slt2p is an enzymatic regulator of silencing function that couples commitment to cell growth and shorter lifespan. PMID- 12640456 TI - Neurotoxic esterase: not so toxic? PMID- 12640457 TI - Hyperpolarization moves S4 sensors inward to open MVP, a methanococcal voltage gated potassium channel. AB - MVP, a Methanococcus jannaschii voltage-gated potassium channel, was cloned and shown to operate in eukaryotic and prokaryotic cells. Like pacemaker channels, MVP opens on hyperpolarization using S4 voltage sensors like those in classical channels activated by depolarization. The MVP S4 span resembles classical sensors in sequence, charge, topology and movement, traveling inward on hyperpolarization and outward on depolarization (via canaliculi in the protein that bring the extracellular and internal solutions into proximity across a short barrier). Thus, MVP opens with sensors inward indicating a reversal of S4 position and pore state compared to classical channels. Homologous channels in mammals and plants are expected to function similarly. PMID- 12640458 TI - Zinc-mediated inhibition of GABA(A) receptors: discrete binding sites underlie subtype specificity. AB - Zinc ions are concentrated in the central nervous system and regulate GABA(A) receptors, which are pivotal mediators of inhibitory synaptic neurotransmission. Zinc ions inhibit GABA(A) receptor function by an allosteric mechanism that is critically dependent on the receptor subunit composition: alphabeta subunit combinations show the highest sensitivity, and alphabetagamma isoforms are the least sensitive. Here we propose a mechanistic and structural basis for this inhibition and its dependence on the receptor subunit composition. We used molecular modeling to identify three discrete sites that mediate Zn2+ inhibition. One is located within the ion channel, and the other two are on the external amino (N)-terminal face of the receptor at the interfaces between alpha and beta subunits. We found that the characteristically low Zn2+ sensitivity of GABA(A) receptors containing the gamma2 subunit results from disruption to two of the three sites after receptor subunit co-assembly. PMID- 12640459 TI - Graspable objects grab attention when the potential for action is recognized. AB - Visually guided grasping movements require a rapid transformation of visual representations into object-specific motor programs. Here we report that graspable objects may facilitate these visuomotor transformations by automatically grabbing visual spatial attention. Human subjects viewed two task irrelevant objects--one was a 'tool', the other a 'non-tool'--while waiting for a target to be presented in one of the two object locations. Using event-related potentials (ERPs), we found that spatial attention was systematically drawn to tools in the right and lower visual fields, the hemifields that are dominant for visuomotor processing. Using event-related fMRI, we confirmed that tools grabbed spatial attention only when they also activated dorsal regions of premotor and prefrontal cortices, regions associated with visually guided actions and their planning. Although it is widely accepted that visual sensory gain aids perception, our results suggest that it may also have consequences for object directed actions. PMID- 12640460 TI - Translation of clock rhythmicity into neural firing in suprachiasmatic nucleus requires mGluR-PLCbeta4 signaling. PMID- 12640461 TI - EphB receptors and ephrin-B ligands regulate spinal sensory connectivity and modulate pain processing. PMID- 12640462 TI - Islet beta-cell secretion determines glucagon release from neighbouring alpha cells. AB - Homeostasis of blood glucose is maintained by hormone secretion from the pancreatic islets of Langerhans. Glucose stimulates insulin secretion from beta cells but suppresses the release of glucagon, a hormone that raises blood glucose, from alpha-cells. The mechanism by which nutrients stimulate insulin secretion has been studied extensively: ATP has been identified as the main messenger and the ATP-sensitive potassium channel as an essential transducer in this process. By contrast, much less is known about the mechanisms by which nutrients modulate glucagon secretion. Here we use conventional pancreas perfusion and a transcriptional targeting strategy to analyse cell-type-specific signal transduction and the relationship between islet alpha- and beta-cells. We find that pyruvate, a glycolytic intermediate and principal substrate of mitochondria, stimulates glucagon secretion. Our analyses indicate that, although alpha-cells, like beta-cells, possess the inherent capacity to respond to nutrients, secretion from alpha-cells is normally suppressed by the simultaneous activation of beta-cells. Zinc released from beta-cells may be implicated in this suppression. Our results define the fundamental mechanisms of differential responses to identical stimuli between cells in a microorgan. PMID- 12640464 TI - PKCepsilon is a permissive link in integrin-dependent IFN-gamma signalling that facilitates JAK phosphorylation of STAT1. AB - The critical dependence of receptor-triggered signals on integrin-mediated cell substrate interactions represents a fundamental biological paradigm in health and disease. However, the molecular connections of these permissive inputs, which operate through integrin-matrix interactions, has remained largely obscure. Here we show that the serine-threonine kinase protein kinase C epsilon (PKCepsilon) functions as a signal integrator between cytokine and integrin signalling pathways. Integrins are shown to control PKCepsilon phosphorylation acutely by determining complex formation with protein phosphatase 2A (PP2A) and the upstream kinase PDK1 (phosphoinositide-dependent kinase 1). The PP2A-induced loss of PKCepsilon function results in attenuated interferon gamma (INF-gamma)-induced phosphorylation of STAT1 (signal transducer and activator of transcription 1) downstream of Janus kinase 1/2 (JAK1/2). PKCepsilon function and the IFN-gamma response can be recovered by inhibition of PP2A if PDK1 is associated with PKCepsilon in this complex. More directly, a PP2A-resistant mutant of PKCepsilon is sufficient for restoration of the IFN-gamma response in suspension culture. Thus, PKCepsilon functions as a central point of integration through which integrin engagement exerts a permissive input on IFN-gamma signalling. PMID- 12640463 TI - Human CENP-I specifies localization of CENP-F, MAD1 and MAD2 to kinetochores and is essential for mitosis. AB - The kinetochore, a macromolecular complex located at the centromere of chromosomes, provides essential functions for accurate chromosome segregation. Kinetochores contain checkpoint proteins that monitor attachments between the kinetochore and microtubules to ensure that cells do not exit mitosis in the presence of unaligned chromosomes. Here we report that human CENP-I, a constitutive protein of the kinetochore that shares limited similarity with Mis6 of Schizosaccharomyces pombe, is required for the localization of CENP-F and the checkpoint proteins MAD1 and MAD2 to kinetochores. Depletion of CENP-I from kinetochores causes the cell cycle to delay in G2. Although monopolar chromosomes in CENP-I-depleted cells fail to establish bipolar connections, the cells are unable to arrest in mitosis. These cells are transiently delayed in mitosis in a MAD2-dependent manner, even though their kinetochores are depleted of MAD2. The delay is extended considerably when the number of unattached kinetochores is increased. This suggests that no single unattached kinetochore in CENP-I-depleted cells can arrest mitosis. The collective output from many unattached kinetochores is required to reach a threshold signal of 'wait for anaphase' to sustain a prolonged mitotic arrest. PMID- 12640465 TI - Semisynthetic production of unnatural L-alpha-amino acids by metabolic engineering of the cysteine-biosynthetic pathway. AB - There is an increasing demand for peptide-mimicking molecules to modulate the interactions between proteins of pharmaceutical and agrochemical interest and their target polypeptides. Unnatural L-alpha-amino acids differing from the 20 naturally proteinogenic amino acids only in their side chain are ideal for this purpose, but their chemical synthesis is complex. Here we describe a fermentation based approach for biosynthesis of unnatural amino acids after re-engineering the cysteine-biosynthetic pathway in Escherichia coli. O-acetylation of serine, the committed step of the pathway, was released from feedback inhibition by mutating the serine acetyltransferase gene. Next, the naturally broad substrate specificity of O-acetylserine sulfhydrylase was exploited for the direct in vivo incorporation of an unnatural side chain in a semisynthetic fermentation process comparable to the production of beta-lactams. O-acetyl-L-serine extruded from the cells by way of the O-acetylserine efflux protein was amenable to further biotransformations. PMID- 12640466 TI - Widespread occurrence of antisense transcription in the human genome. AB - An increasing number of eukaryotic genes are being found to have naturally occurring antisense transcripts. Here we study the extent of antisense transcription in the human genome by analyzing the public databases of expressed sequences using a set of computational tools designed to identify sense-antisense transcriptional units on opposite DNA strands of the same genomic locus. The resulting data set of 2,667 sense-antisense pairs was evaluated by microarrays containing strand-specific oligonucleotide probes derived from the region of overlap. Verification of specific cases by northern blot analysis with strand specific riboprobes proved transcription from both DNA strands. We conclude that > or =60% of this data set, or approximately 1,600 predicted sense-antisense transcriptional units, are transcribed from both DNA strands. This indicates that the occurrence of antisense transcription, usually regarded as infrequent, is a very common phenomenon in the human genome. Therefore, antisense modulation of gene expression in human cells may be a common regulatory mechanism. PMID- 12640467 TI - Multiherbicide tolerance conferred by AtPgp1 and apyrase overexpression in Arabidopsis thaliana. AB - Herbicide resistance is an important trait often introduced into crop plants. Mechanisms of resistance can involve a mutant target protein that is unaffected by the herbicide, or metabolic detoxification or degradation of the herbicide. Recently, we showed that overexpression in Arabidopsis thaliana of either psNTP9, the garden pea apyrase gene, or AtPgp1, the A. thaliana homolog of the plant multidrug resistance (MDR) gene, enabled A. thaliana to germinate on the toxin cycloheximide and to grow better on toxic levels of the plant hormone N6-[2 isopentyl]adenine (2iP). Here we report that overexpression of either MDR or apyrase proteins resulted in increased resistance to herbicides from different chemical classes. Apyrase inhibition by small molecule inhibitors reversed this resistance. Treatment of untransformed plants with an apyrase inhibitor increased their sensitivity to the same herbicides. These results indicate that the genes may be involved in a resistance mechanism relating to decreased retention or increased active efflux of herbicide from the plant cell. PMID- 12640468 TI - New directions for WHO? PMID- 12640469 TI - Compatible poliomyelitis cases in India during 2000. AB - OBJECTIVE: To describe the characteristics of compatible poliomyelitis cases and to assess the programmatic implications of clusters of such cases in India. METHODS: We described the characteristics of compatible poliomyelitis cases, identified clusters of compatible cases (two or more in the same district or neighbouring districts within two months), and examined their relationship to wild poliovirus cases. FINDINGS: There were 362 compatible cases in 2000. The incidence of compatible cases was higher in districts with laboratory-confirmed poliomyelitis cases than in districts without laboratory-confirmed cases. Of 580 districts, 96 reported one compatible case and 72 reported two or more compatible cases. Among these 168 districts with at least one compatible case, 123 had internal or cross- border clusters of compatible cases. In 27 districts with clusters of compatible cases, no wild poliovirus was isolated either in the same district or in neighbouring districts. Three of these 27 districts presented laboratory-confirmed poliomyelitis cases during 2001. CONCLUSION: Most clusters of compatible cases occurred in districts identified as areas with continuing wild poliovirus transmission and where mopping-up vaccination campaigns were carried out. As certification nears, areas with compatible poliomyelitis cases should be investigated and deficiencies in surveillance should be corrected in order to ensure that certification is justified. PMID- 12640470 TI - Violence against women increases the risk of infant and child mortality: a case referent study in Nicaragua. AB - OBJECTIVE: To investigate the impact of violence against mothers on mortality risks for their offspring before 5 years of age in Nicaragua. METHODS: From a demographic database covering a random sample of urban and rural households in Le n, Nicaragua, we identified all live births among women aged 15-49 years. Cases were defined as those who had died before the age of 5 years, between January 1993 and June 1996. For each case, two referents, matched for sex and age at death, were selected from the database. A total of 110 mothers of the cases and 203 mothers of the referents were interviewed using a standard questionnaire covering mothers' experience of physical and sexual violence. The data were analysed for the risk associated with maternal experience of violence of infant and under-5 mortality. FINDINGS: A total of 61% of mothers of cases had a lifetime experience of physical and/or sexual violence compared with 37% of mothers of referents, with a significant association being found between such experiences and mortality among their offspring. Other factors associated with higher infant and under-5 mortality were mother's education (no formal education), age (older), and parity (multiparity). CONCLUSIONS: The results suggest an association between physical and sexual violence against mothers, either before or during pregnancy, and an increased risk of under-5 mortality of their offspring. The type and severity of violence was probably more relevant to the risk than the timing, and violence may impact child health through maternal stress or care-giving behaviours rather than through direct trauma itself. PMID- 12640471 TI - Violence against women and the risk of infant and child mortality. PMID- 12640472 TI - The cost of diabetes in Latin America and the Caribbean. AB - OBJECTIVE: To measure the economic burden associated with diabetes mellitus in Latin America and the Caribbean. METHODS: Prevalence estimates of diabetes for the year 2000 were used to calculated direct and indirect costs of diabetes mellitus. Direct costs included costs due to drugs, hospitalizations, consultations and management of complications. The human capital approach was used to calculate indirect costs and included calculations of forgone earnings due to premature mortality and disability attributed to diabetes mellitus. Mortality and disability attributed to causes other than diabetes were subtracted from estimates to consider only the excess burden due to diabetes. A 3% discount rate was used to convert future earnings to current value. FINDINGS: The annual number of deaths in 2000 caused by diabetes mellitus was estimated at 339,035. This represented a loss of 757,096 discounted years of productive life among persons younger than 65 years (> billion US dollars). Permanent disability caused a loss of 12,699,087 years and over 50 billion US dollars, and temporary disability caused a loss of 136,701 years in the working population and over 763 million US dollars. Costs associated with insulin and oral medications were 4720 million US dollars, hospitalizations 1012 million US dollars, consultations 2508 million US dollars and care for complications 2,480 million US dollars. The total annual cost associated with diabetes was estimated as 65,216 million US dollars (direct 10,721 US dollars; indirect 54,496 US dollars). CONCLUSION: Despite limitations of the data, diabetes imposes a high economic burden to individuals and society in all countries and to Latin American and the Caribbean as whole. PMID- 12640473 TI - Prevalence of goitre and urinary iodine status of primary-school children in Lesotho. AB - OBJECTIVE: To estimate the prevalence of goitre, urinary iodine status, coverage of supplementation of iodized oil capsules, and current use of iodized salt in children in Lesotho. METHODS: Cross-sectional study of children from 50 primary schools in Lesotho. Thyroid glands of children aged 8-12 years were measured by palpation and graded according to the WHO, UNICEF, and the International Council for the Control of Iodine Deficiency's (ICCIDD) joint criteria. The use of iodized oil capsules was determined by a structured questionnaire and verified with the children's health booklets. Iodine content of household salt samples was analysed. Casual urine samples were analysed for urinary iodine. FINDINGS: Median urinary iodine concentrations of 26.3 microg/l (range 22.3-47.9 microg/l) indicated moderate iodine deficiency. More children in the mountains than in the lowlands were severely iodine deficient (17.7% vs 1.9%). Adjusted prevalence of goitre (4.9%) increased with age, was higher in girls than boys, and ranged from 2.2% to 8.8% in the different districts; this indicated no public health problem. Overall, 94.4% of salt samples were iodized, and coverage of supplementation with iodized oil capsules was 55.1%. CONCLUSION: Mild-to-moderate iodine deficiency exists in Lesotho. Iodine deficiency was more severe in the mountains than the lowlands and is still a concern for public health. Use of iodized salt coupled with iodized oil supplementation effectively controls iodine deficiency disorders. Effective monitoring programmes would ensure the use of adequately iodized salt throughout Lesotho and serve to evaluate progress towards optimal iodine nutrition. Iodized oil capsule supplementation should continue in the mountains. PMID- 12640474 TI - A comparison of the efficacy of single doses of albendazole, ivermectin, and diethylcarbamazine alone or in combinations against Ascaris and Trichuris spp. AB - OBJECTIVE: To determine the efficacy of single doses of albendazole, ivermectin and diethylcarbamazine, and of the combinations albendazole + ivermectin and albendazole + diethylcarbamazine against common intestinal helminthiases caused by Ascaris and Trichuris spp. METHODS: In a randomized, placebo-controlled trial, infected children were randomly assigned to treatment with albendazole + placebo, ivermectin + placebo, diethylcarbamazine + placebo, albendazole + ivermectin, or albendazole + diethylcarbamazine. The Kato-Katz method was used for qualitative and quantitative parasitological diagnosis. The chi2 test was used to determine the significance of cure rates, repeated measures analysis of variance for the comparison of mean log egg counts, the Newman-Keuls procedure for multiple comparison tests, and logistic regression for the comparison of infection rates at days 180 and 360 after treatment. FINDINGS: Albendazole, ivermectin and the drug combinations gave significantly higher cure and egg reduction rates for ascariasis than diethylcarbamazine. For trichuriasis, albendazole + ivermectin gave significantly higher cure and egg reduction rates than the other treatments: the infection rates were lower 180 and 360 days after treatment. CONCLUSION: Because of the superiority of albendazole + ivermectin against both lymphatic filariasis and trichuriasis, this combination appears to be a suitable tool for the integrated or combined control of both public health problems. PMID- 12640475 TI - Healthy life expectancy in Hong Kong Special Administrative Region of China. AB - Sullivan's method and a regression model were used to calculate healthy life expectancy (HALE) for men and women in Hong Kong Special Administrative Region (Hong Kong SAR) of China. These methods need estimates of the prevalence and information on disability distributions of 109 diseases and HALE for 191 countries by age, sex and region of the world from the WHO's health assessment of 2000. The population of Hong Kong SAR has one of the highest healthy life expectancies in the world. Sullivan's method gives higher estimates than the classic linear regression method. Although Sullivan's method accurately calculates the influence of disease prevalence within small areas and regions, the regression method can approximate HALE for all economies for which information on life expectancy is available. This paper identifies some problems of the two methods and discusses the accuracy of estimates of HALE that rely on data from the WHO assessment. PMID- 12640476 TI - Tobacco control in India. AB - Legislation to control tobacco use in developing countries has lagged behind the dramatic rise in tobacco consumption. India, the third largest grower of tobacco in the world, amassed 1.7 million disability-adjusted life years (DALYs) in 1990 due to disease and injury attributable to tobacco use in a population where 65% of the men and 38% of the women consume tobacco. India's anti-tobacco legislation, first passed at the national level in 1975, was largely limited to health warnings and proved to be insufficient. In the last decade state legislation has increasingly been used but has lacked uniformity and the multipronged strategies necessary to control demand. A new piece of national legislation, proposed in 2001, represents an advance. It includes the following key demand reduction measures: outlawing smoking in public places; forbidding sale of tobacco to minors; requiring more prominent health warning labels; and banning advertising at sports and cultural events. Despite these measures, the new legislation will not be enough to control the demand for tobacco products in India. The Indian Government must also introduce policies to raise taxes, control smuggling, close advertising loopholes, and create adequate provisions for the enforcement of tobacco control laws. PMID- 12640477 TI - Domestic violence in rural Uganda: evidence from a community-based study. AB - Although domestic violence is an increasing public health concern in developing countries, evidence from representative, community-based studies is limited. In a survey of 5109 women of reproductive age in the Rakai District of Uganda, 30% of women had experienced physical threats or physical abuse from their current partner--20% during the year before the survey. Three of five women who reported recent physical threats or abuse reported three or more specific acts of violence during the preceding year, and just under a half reported injuries as a result. Analysis of risk factors highlights the pivotal roles of the male partner's alcohol consumption and his perceived human immunodeficiency virus (HIV) risk in increasing the risk of male against female domestic violence. Most respondents- 70% of men and 90% of women--viewed beating of the wife or female partner as justifiable in some circumstances, posing a central challenge to preventing violence in such settings. PMID- 12640478 TI - Immunization of children at risk of infection with human immunodeficiency virus. AB - This paper reviews the English language literature on the safety, immunogenicity and effectiveness in children infected with the human immunodeficiency virus (HIV) of vaccines currently recommended by WHO for use in national immunization programmes. Immunization is generally safe and beneficial for children infected with HIV, although HIV-induced immune suppression reduces the benefit compared with that obtained in HIV-uninfected children. However, serious complications can occur following immunization of severely immunocompromised children with bacillus Calmette-Gu rin (BCG) vaccine. The risk of serious complications attributable to yellow fever vaccine in HIV-infected persons has not been determined. WHO guidelines for immunizing children with HIV infection and infants born to HIV infected women differ only slightly from the general guidelines. BCG and yellow fever vaccines should be withheld from symptomatic HIV-infected children. Only one serious complication (fatal pneumonia) has been attributed to measles vaccine administered to a severely immunocompromised adult. Although two HIV-infected infants have developed vaccine-associated paralytic poliomyelitis, several million infected children have been vaccinated and the evidence does not suggest that there is an increased risk. The benefits of measles and poliovirus vaccines far outweigh the potential risks in HIV-infected children. The policy of administering routine vaccines to all children, regardless of possible HIV exposure, has been very effective in obtaining high immunization coverage and control of preventable diseases. Any changes in this policy would have to be carefully examined for a potential negative impact on disease control programmes in many countries. PMID- 12640479 TI - New organization to promote African health research. PMID- 12640480 TI - In South Africa HIV infection is decreasing, safe sex increasing. PMID- 12640481 TI - Drug companies should cut prices for developing countries, says G8 report. PMID- 12640483 TI - Peru tries vinegar against cervical cancer. PMID- 12640484 TI - Is breast cancer linked to smoking? PMID- 12640491 TI - Effect of lung resection and sham surgery on the frequency of infection in alloxan-diabetic rats. AB - The present study was carried out in order to determine the effect of lung resection on the frequency of infections in alloxan-diabetic rats. Adult female Wistar rats were injected with alloxan (40 mg/kg, iv) to induce diabetes mellitus (group D; N = 45) or with vehicle (1.0 ml/kg, iv) to be used as controls (group C; N = 45). Thirty-six days after receiving alloxan both groups were randomly divided into three subgroups: no operation (NO; N = 15), sham operation (SO; N = 15), and left pneumonectomy (PE; N = 15). The rats were sacrificed 36 days after surgery and their lungs were examined microscopically and macroscopically. The occurrence of thoracic wall infection, thoracic wall abscess, lung abscess and pleural empyema was similar in groups D and C. In contrast, the overall infection rate was higher (P<0.05) in the diabetic rats (SO-D and PE-D subgroups, but not in the NO-D subgroup). Considering that the overall infection rate was similar in the SO-D and PE-D subgroups, we suggest that surgery but not pneumonectomy was related to the higher prevalence of infection in diabetic rats. PMID- 12640492 TI - Pulmonary function, cholinergic bronchomotor tone, and cardiac autonomic abnormalities in type 2 diabetic patients. AB - This prospective study analyzed the involvement of the autonomic nervous system in pulmonary and cardiac function by evaluating cardiovascular reflex and its correlation with pulmonary function abnormalities of type 2 diabetic patients. Diabetic patients (N = 17) and healthy subjects (N = 17) were evaluated by 1) pulmonary function tests including spirometry, He-dilution method, N2 washout test, and specific airway conductance (SGaw) determined by plethysmography before and after aerosol administration of atropine sulfate, and 2) autonomic cardiovascular activity by the passive tilting test and the magnitude of respiratory sinus arrhythmia (RSA). Basal heart rate was higher in the diabetic group (87.8 +/- 11.2 bpm; mean +/- SD) than in the control group (72.9 +/- 7.8 bpm, P<0.05). The increase of heart rate at 5 s of tilting was 11.8 +/- 6.5 bpm in diabetic patients and 17.6 +/- 6.2 bpm in the control group (P<0.05). Systemic arterial pressure and RSA analysis did not reveal significant differences between groups. Diabetes intragroup analysis revealed two behaviors: 10 patients with close to normal findings and 7 with significant abnormalities in terms of RSA, with the latter subgroup presenting one or more abnormalities in other tests and clear evidence of cardiovascular autonomic dysfunction. End-expiratory flows were significantly lower in diabetic patients than in the control group (P<0.05). Pulmonary function tests before and after atropine administration demonstrated comparable responses by both groups. Type 2 diabetic patients have cardiac autonomic dysfunction that is not associated with bronchomotor tone alterations, probably reflecting a less severe impairment than that of type 1 diabetes mellitus. Yet, a reduction of end-expiratory flow was detected. PMID- 12640493 TI - Insulin secretion, insulin sensitivity, and hepatic insulin extraction in first degree relatives of type 2 diabetic patients. AB - To identify early metabolic abnormalities in type 2 diabetes mellitus, we measured insulin secretion, sensitivity to insulin, and hepatic insulin extraction in 48 healthy normal glucose-tolerant Brazilians, first-degree relatives of type 2 diabetic patients (FH+). Each individual was matched for sex, age, weight, and body fat distribution with a person without history of type 2 diabetes (FH-). Both groups were submitted to a hyperglycemic clamp procedure (180 mg/dl). Insulin release was evaluated in its two phases. The first was calculated as the sum of plasma insulin at 2.5, 5.0, 7.5, and 10.0 min after the beginning of glucose infusion, and the second as the mean plasma insulin level in the third hour of the clamp procedure. Insulin sensitivity index (ISI) was the mean glucose infusion rate in the third hour of the clamp experiment divided by the mean plasma insulin concentration during the same period of time. Hepatic insulin extraction was determined under fasting conditions and in the third hour of the clamp procedure as the ratio between C-peptide and plasma insulin levels. FH+ individuals did not differ from FH- individuals in terms of the following parameters [median (range)]: a) first-phase insulin secretion, 174 (116-221) vs 207 (108-277) microU/ml, b) second-phase insulin secretion, 64 (41-86) vs 53 (37 83) microU/ml, and c) ISI, 14.8 (9.0-20.8) vs 16.8 (9.0-27.0) mg kg-1 min-1/ microU ml-1. Hepatic insulin extraction in FH+ subjects was similar to that of FH ones at basal conditions (median, 0.27 vs 0.27 ng/microU) and during glucose infusion (0.15 vs 0.15 ng/ micro U). Normal glucose-tolerant Brazilian FH+ individuals well-matched with FH- ones did not show defects of insulin secretion, insulin sensitivity, or hepatic insulin extraction as tested by hyperglycemic clamp procedures. PMID- 12640494 TI - The calcium-dependent protease of Loxosceles gaucho venom acts preferentially upon red cell band 3 transmembrane protein. AB - Eighty micrograms red blood cell (RBC) ghosts from patients who had previously exhibited the cutaneous form of loxoscelism (presenting localized dermonecrosis) and the viscerocutaneous form of loxoscelism (presenting dermonecrosis, hemoglobinuria, hematuria, and jaundice) and from controls were incubated with 2.5 microg crude Loxosceles gaucho venom in 5 mM phosphate buffer, pH 7.4, at 37 C. Among all membrane proteins, quantitative proteolysis of the important integral transmembrane protein 3 increased with venom dose and with incubation time from 30 to 120 min, as demonstrated by gel densitometry. Similar quantitative data were obtained for RBC ghosts from patients and from control subjects, a fact that argues against the possibility of genetic factors favoring the hemolytic viscerocutaneous form. These data suggest that the clinical forms may be different types of the same disease, with the viscerocutaneous form being the result of large amounts of intravascularly injected venom and the superficial form being the result of in situ venom action. Since protein 3 is a housekeeping integral membrane protein, whose genetic deficiency leads to hemolytic anemia, it is reasonable to relate it to the hemolysis which occurs in the viscerocutaneous form of loxoscelism. The venom protease responsible for the process was not inhibited after 120-min incubation by 0.2 mM paramethylsulfonyl fluoride or by 0.2 mM N-ethylmaleimide but was inhibited by 25 mM ethylenediaminetetraacetic acid (a calcium-chelating agent) in 5 mM phosphate buffer at pH 7.4, which suggests that the enzyme is a calcium-dependent metalloprotease. PMID- 12640495 TI - The availability of full match sibling donors and feasibility of allogeneic bone marrow transplantation in Brazil. AB - The feasibility of allogeneic bone marrow transplantation (alloBMT) in a developing country has not yet been demonstrated. Many adverse factors including social and economic limitations may reduce the overall results of this complex and expensive procedure. Our objective was to characterize the most important clinical, social and economic features of candidates for transplantation and their potential donors as well as the influence of these factors on overall survival in a retrospective and exploratory analysis at a university hospital. From July 1993 to July 2001, candidates for BMT were referred to the Bone Marrow Transplantation Unit by Hematology and Oncology Centers from several regions of Brazil. A total of 1138 patients were referred to us as candidates for alloBMT. Median age was 25 years (range: 2 months-60 years), 684 (60.1%) were males and 454 (39.9%) were females. The clinical indications were severe aplastic anemia and hematological malignancies. From the total of 1138 patients, 923 had HLA typing; 497/923 (53.8%) candidates had full match donors; 352/1138 (30.8%) were eligible for alloBMT. Only 235 of 352 (66.7%) were transplanted. Schooling was 1st to 8th grade for 123/235 (52.3%); monthly family income ranged from US$60 (7%) to more than US$400 (36%). Overall survival for patients with chronic myeloid leukemia, severe aplastic anemia and acute myeloid leukemia was 58, 60 and 30%, respectively. Thus, overall survival rates for the most frequent hematological diseases were similar to those reported in the International Registry, except for acute myeloid leukemia. This descriptive and exploratory analysis suggests the feasibility of alloBMT in a developing country like Brazil. PMID- 12640496 TI - Hepatitis C virus infection in a Brazilian population with sickle-cell anemia. AB - Patients with sickle-cell anemia submitted to frequent blood transfusions are at risk of contamination with hepatitis C virus (HCV). Determination of HCV RNA and genotype characterization are parameters that are relevant for the treatment of the viral infection. The objective of the present study was to determine the frequency of HCV infection and the positivity for HCV RNA and to identify the HCV genotype in patients with sickle-cell anemia with a history of blood transfusion who had been treated at the Hospital of the HEMOPE Foundation. Sera from 291 patients were tested for anti-HCV antibodies by ELISA 3.0 and RIBA 3.0 Chiron and for the presence of HCV RNA by RT-PCR. HCV genotyping was performed in 19 serum samples. Forty-one of 291 patients (14.1%) were anti-HCV positive by ELISA and RIBA. Both univariate and multivariate analysis showed a greater risk of anti-HCV positivity in those who had started a transfusion regime before 1992 and received more than 10 units of blood. Thirty-four of the anti-HCV-positive patients (34/41, 82.9%) were also HCV RNA positive. Univariate analysis, used to compare HCV RNA-negative and -positive patients, did not indicate a higher risk of HCV RNA positivity for any of the variables evaluated. The genotypes identified were 1b (63%), 1a (21%) and 3a (16%). A high prevalence of HCV infection was observed in our patients with sickle-cell anemia (14.1%) compared to the population in general (3%). In the literature, the frequency of HCV infection in sickle-cell anemia ranges from 2 to 30%. The serological screening for anti-HCV at blood banks after 1992 has contributed to a better control of the dissemination of HCV infection. Because of the predominance of genotype 1, these patients belong to a group requiring special treatment, with a probable indication of new therapeutic options against HCV. PMID- 12640497 TI - Acute leukemias in Piaui: comparison with features observed in other regions of Brazil. AB - Differences in age and sex distribution as well as FAB (French-American-British classification) types have been reported for acute leukemias in several countries. We studied the demographics and response to treatment of patients with acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) between 1989 and 2000 in Teresina, Piau , and compared these results with reports from Brazil and other countries. Complete data concerning 345 patients (230 ALL, 115 AML) were reviewed. AML occurred predominantly in adults (77%), with a median age of 34 years, similar to that found in the southeast of Brazil but lower than the median age in the United States and Europe (52 years). FAB distribution was similar in children and adults and FAB-M2 was the most common type, as also found in Japan. The high frequency of FAB-M3 described in most Brazilian studies and for Hispanics in the United States was not observed. Overall survival for adults was 40%, similar to other studies in Brazil. A high mortality rate was observed during induction. No clinical or hematological parameter influenced survival in the Cox model. ALL presented the characteristic peak of incidence between 2-8 years. Most of the cases were CD10+ pre-B ALL. In 25%, abnormal expression of myeloid antigens was observed. Only 10% of the patients were older than 30 years. Overall survival was better for children. Age and leukocyte count were independent prognostic factors. These data demonstrate that, although there are regional peculiarities, the application of standardized treatments and good supportive care make it possible to achieve results observed in other countries for the same chemotherapy protocols. PMID- 12640498 TI - Effect of surgical treatment on the cellular immune response of gastric cancer patients. AB - Patients with gastric cancer have a variety of immunological abnormalities. In the present study the lymphocytes and their subsets were determined in the peripheral blood of patients with gastric cancer (N = 41) both before and after surgical treatment. The percent of helper/inducer CD4 T cells (43.6 +/- 8.9) was not different after tumor resection (43.6 +/- 8.2). The percent of the cytotoxic CD8+ T cell population decreased significantly, whether patients were treated surgically (27.2 +/- 5.8%, N = 20) or not (27.3 +/- 7.3%, N = 20) compared to individuals with inflammatory disease (30.9 +/- 7.5%) or to healthy individuals (33.2 +/- 7.6%). The CD4/CD8 ratio consequently increased in the group of cancer patients. The peripheral blood lymphocytes of gastric cancer patients showed reduced responsiveness to mitogens. The defective blastogenic response of the lymphocytes was not associated with the production of transforming growth factor beta (TGF- ) since the patients with cancer had reduced production of TGF- Beta1 (269 +/- 239 pg/ml, N = 20) in comparison to the normal individuals (884 +/- 175 pg/ml, N = 20). These results indicate that the immune response of gastric cancer patients was not significantly modified by surgical treatment when evaluated four weeks after surgery and that the immunosuppression observed was not due to an increase in TGF- 1 production by peripheral leukocytes. PMID- 12640499 TI - Vertical growth phase and positive sentinel node in thin melanoma. AB - Sentinel node (SN) status is the most important prognostic factor for localized melanoma. Usually, patients with Breslow thickness of less than 1.0 mm are not included in SN protocols. However, the literature presents a rate ranging from 3 to 7% of nodal recurrence in thin melanoma. Ulceration, regression and high mitotic rate have been considered to be indications for an SN biopsy. The metastatic potential of the vertical growth phase is uncertain. To correlate pathological features in thin melanoma with SN metastasis, we reviewed 358 patients submitted to SN biopsy. Seventy-seven patients with lesions of 1 mm or smaller were included in the study group. Histological evaluation of the primary tumor included thickness, Clark level, mitotic rate, ulceration, regression, and growth phase. Lymphoscintigraphy was performed on all patients. Lymphatic mapping and gamma probe detection were both used for SN biopsy. Histological examination of SN consisted of hematoxylin-eosin and immunohistochemical staining. Median follow-up was 37 months. Six patients had micrometastases. Statistical analysis by the Fisher test showed that ulceration (P = 0.019), high mitotic rate (P = 0.008) and vertical growth phase (P = 0.002) were positively correlated with micrometastases. If other studies confirm these results, more melanoma patients must be submitted to SN biopsy. PMID- 12640500 TI - Biological activity of Serratia marcescens cytotoxin. AB - Serratia marcescens cytotoxin was purified to homogeneity by ion-exchange chromatography on a DEAE Sepharose Fast Flow column, followed by gel filtration chromatography on a Sephadex G100 column. The molecular mass of the cytotoxin was estimated to be about 50 kDa. Some biological properties of the cytotoxin were analyzed and compared with well-characterized toxins, such as VT1, VT2 and CNF from Escherichia coli and hemolysin produced by S. marcescens. The sensitivity of the cell lines CHO, HeLa, HEp-2, Vero, BHK-21, MA 104 and J774 to the cytotoxin was determined by the cell viability assay using neutral red. CHO and HEp-2 were highly sensitive, with massive cellular death after 1 h of treatment, followed by BHK-21, HeLa, Vero and J774 cells, while MA 104 was insensitive to the toxin. Cytotoxin induced morphological changes such as cell rounding with cytoplasmic retraction and nuclear compactation which were evident 15 min after the addition of cytotoxin. The cytotoxic assays show that 15 min of treatment with the cytotoxin induced irreversible intoxication of the cells, determined by loss of cell viability. Concentrations of 2 CD50 (0.56 g/ml) of purified cytotoxin did not present any hemolytic activity, showing that the cytotoxin is distinct from S. marcescens hemolysin. Antisera prepared against S. marcescens cytotoxin did not neutralize the cytotoxic activity of VT1, VT2 or CNF toxin, indicating that these toxins do not share antigenic determinants with cytotoxin. Moreover, we did not detect gene sequences for any of these toxins in S. marcescens by PCR assay. These results suggest that S. marcescens cytotoxin is not related to any of these toxins from E. coli. PMID- 12640501 TI - In vitro-induced antibody production in chronic hepatitis C virus infection. AB - The objectives of the present study were to assess the in vitro-induced anti hepatitis C virus (HCV) antibody production (IVIAP) in relation to the clinical, biochemical, virologic and histologic variables of patients with HCV infection. The study included 57 patients (60% males) with HCV infection (anti-HCV and HCV RNA positive). Alanine aminotransferase (ALT) was elevated in 89% of the patients. Mean viral load was 542,241 copies/ml and histology of the liver showed chronic hepatitis in 27/52 (52%) and cirrhosis in 11/52 (21%) patients. IVIAP levels were determined by immunoenzymatic assay at median absorbance of 0.781 at 450 nm. IVIAP was negative in 14% of the patients. When groups with IVIAP levels above and below the median were compared, high IVIAP levels were associated with the male sex, elevated ALT levels and more advanced disease stage. After logistic regression analysis, advanced histologic damage to the liver remained as the only independent variable associated with elevated IVIAP levels. Using a receiver operator characteristic curve, the best cut-off level for IVIAP was established (= 1.540), with 71% sensitivity and 94% specificity for the detection of more advanced disease stages (grades 3 and 4). These findings are consistent with the participation of immunological mechanisms in the genesis of the hepatic lesions induced by HCV and indicate that the IVIAP test may be useful as a noninvasive marker of liver damage either alone or in combination with other markers. PMID- 12640502 TI - The X-X-/E+E+ genotype of the XbaI/EcoRI polymorphisms of the apolipoprotein B gene as a marker of coronary artery disease in a Brazilian sample. AB - Studies that consider polymorphisms within the apolipoprotein B (apo B) gene as risk factors for coronary artery disease (CAD) have reported conflicting results. The aim of the present study was to search for associations between two DNA RFLPs (XbaI and EcoRI) of the apo B gene and CAD diagnosed by angiography. In the present study we compared 116 Brazilian patients (92 men) with CAD (CAD+) to 78 control patients (26 men) without ischemia or arterial damage (CAD-). The allele frequencies at the XbaI (X) and EcoRI (E) sites did not differ between groups. The genotype distributions of CAD+ and CAD- patients were different (chi (1) = 6.27, P = 0.012) when assigned to two classes (X-X-/E+E+ and the remaining XbaI/EcoRI genotypes). Multivariate logistic regression analysis showed that individuals with the X-X-/E+E+ genotype presented a 6.1 higher chance of developing CAD than individuals with the other XbaI/EcoRI genotypes, independently of the other risk factors considered (sex, tobacco consumption, total cholesterol, hypertension, and triglycerides). We conclude that the X-X /E+E genotype may be in linkage disequilibrium with an unknown variation in the apo B gene or with a variation in another gene that affects the risk of CAD. PMID- 12640503 TI - Serum hexosaminidase and beta-glucuronidase activities in infants: effects of age and sex. AB - We investigated the effect of age and sex on the serum activity of hexosaminidase (HEX) and -glucuronidase (BGLU) in 275 normal term infants aged 12 h to 12 months. Up to six weeks of life, HEX was significantly higher in boys (P<=0.023). During the age period of 1-26 weeks, BGLU was also higher in boys, but differences were significant only at 2-6 and 7-15 weeks (P<=0.016). The developmental pattern of HEX and BGLU was sex dependent. HEX activity increased in both sexes from 4-7 days of life, reaching a maximum of 1.4-fold the birth value at 2-6 weeks of age in boys (P<0.001) and a maximum of 1.6-fold at 7-15 weeks in girls (P<0.001). HEX activity gradually decreased thereafter, reaching significantly lower levels at 27-53 weeks than during the first three days of life in boys (P = 0.002) and the same level of this age interval in girls. BGLU increased in both sexes from 4-7 days of age, showing a maximum increase at 7-15 weeks (3.3-fold in boys and 2.9-fold in girls, both P<0.001). Then BGLU decreased in boys to a value similar to that observed at 4-7 days of age. In girls, BGLU remained elevated until the end of the first year of life. These results indicate a variation of HEX and BGLU activities during the first year of life and a sex influence on their developmental pattern. This observation should be considered in the diagnosis of GM2 gangliosidosis and mucopolysaccharidosis type VII. PMID- 12640504 TI - Discrepancies between clinical diagnoses and autopsy findings. AB - Autopsy examination is considered to be an essential element for medical auditing and teaching. Despite the significant progress in diagnostic procedures, autopsy has not always confirmed the clinical diagnosis. In the present study, we compared the diagnosis recorded on medical charts with reports of 96 autopsies performed at the University Teaching Hospital of the Faculdade de Medicina de Botucatu, Botucatu, SP, Brazil, between 1975 and 1982, and of 156 autopsies performed at the same institution between 1992 and 1996. The clinical diagnosis of the basic cause of death was confirmed at autopsy in 77% of cases. The percent confirmation fell to 60% when the immediate terminal cause of death was considered, and in 25% of cases, the terminal cause was only diagnosed at autopsy. The discrepancies between clinical and autopsy diagnosis were even larger for secondary diagnoses: 50% of them were not suspected upon clinical diagnosis. Among them, we emphasize the diagnosis of venous thromboses (83%), pulmonary embolisms (80%), bronchopneumonias (46%) and neoplasias (38%). Iatrogenic injuries were very frequent, and approximately 90% of them were not described in clinical reports. Our results suggest that highly sensitive and specific diagnostic tests are necessary but cannot substitute the clinical practice for the elaboration of correct diagnoses. PMID- 12640505 TI - Detection of renal dysfunction based on serum creatinine levels in a Brazilian community: the Bambui Health and Ageing Study. AB - There are few population-based studies of renal dysfunction and none conducted in developing countries. In the present study the prevalence and predictors of elevated serum creatinine levels (SCr > or = 1.3 mg/dl for men and 1.1 mg/dl for women) were determined among Brazilian adults (18-59 years) and older adults (>60 years). Participants included all older adults (N = 1742) and a probabilistic sample of adults (N = 818) from Bambu town, MG, Southeast Brazil. Predictors were investigated using multiple logistic regression. Mean SCr levels were 0.77 +/- 0.15 mg/dl for adults, 1.02 +/- 0.39 mg/dl for older men, and 0.81 +/- 0.17 mg/dl for older women. Because there were only 4 cases (0.48%) with elevated SCr levels among adults, the analysis of elevated SCr levels was restricted to older adults. The overall prevalence of elevated SCr levels among the elderly was 5.09% (76/1494). The prevalence of hypercreatinemia increased significantly with age (chi = 26.17, P = 0.000), being higher for older men (8.19%) than for older women (5.29%, chi = 5.00, P = 0.02). Elevated SCr levels were associated with age 70-79 years (odds ratio [OR] = 2.25, 95% confidence interval [CI]: 1.15-4.42), hypertension (OR = 3.04, 95% CI: 1.34-6.92), use of antihypertensive drugs (OR = 2.46, 95% CI: 1.26-4.82), chest pain (OR = 3.37, 95% CI: 1.31-8.74), and claudication (OR = 3.43, 95% CI: 1.30-9.09) among men, and with age >80 years (OR = 4.88, 95% CI: 2.24-10.65), use of antihypertensive drugs (OR = 4.06, 95% CI: 1.67-9.86), physical inactivity (OR = 2.11, 95% CI: 1.11-4.02) and myocardial infarction (OR = 3.89, 95% CI: 1.58-9.62) among women. The prevalence of renal dysfunction observed was much lower than that reported in other population-based studies, but predictors were similar. New investigations are needed to confirm the variability in prevalence and associated factors of renal dysfunction among populations. PMID- 12640506 TI - White-coat hypertension and normotension in the League of Hypertension of the Hospital das Clinicas, FMUSP: prevalence, clinical and demographic characteristics. AB - OBJECTIVE: To assess the prevalence of white-coat normortension, white-coat hypertension, and white-coat effect. METHODS: We assessed 670 medical records of patients from the League of Hypertension of the Hospital das Clinicas of the Medical School of the University of S o Paulo. White-coat hypertension (blood pressure at the medical office: mean of 3 measurements with the oscillometric device > or = 140 or > or = 90 mmHg, or both, and ambulatory blood pressure monitoring mean during wakefulness < 135/85) and white-coat normotension (office blood pressure < 140/90 and blood pressure during wakefulness on ambulatory blood pressure monitoring > or = 135/85) were analyzed in 183 patients taking no medication. The white-coat effect (difference between office and ambulatory blood pressure > 20 mmHg for systolic and 10 mmHg for diastolic) was analyzed in 487 patients on treatment, 374 of whom underwent multivariate analysis to identify the variables that better explain the white-coat effect. RESULTS: Prevalence of white-coat normotension was 12%, prevalence of white-coat hypertension was 20%, and prevalence of the white-coat effect was 27%. A significant correlation (p<0.05) was observed between white-coat hypertension and familial history of hypertension, and between the white-coat effect and sex, severity of the office diastolic blood pressure, and thickness of left ventricular posterior wall. CONCLUSION: White-coat hypertension, white-coat normotension, and white-coat effect should be considered in the diagnosis of hypertension. PMID- 12640507 TI - Exercise testing in hypertensive patients taking different angiotensin-converting enzyme inhibitors. AB - OBJECTIVE: To compare blood pressure response to dynamic exercise in hypertensive patients taking trandolapril or captopril. METHODS: We carried out a prospective, randomized, blinded study with 40 patients with primary hypertension and no other associated disease. The patients were divided into 2 groups (n=20), paired by age, sex, race, and body mass index, and underwent 2 symptom-limited exercise tests on a treadmill before and after 30 days of treatment with captopril (75 to 150 mg/day) or trandolapril (2 to 4 mg/day). RESULTS: The groups were similar prior to treatment (p<0.05), and both drugs reduced blood pressure at rest (p<0.001). During treatment, trandolapril caused a greater increase in functional capacity (+31%) than captopril (+17%; p=0.01) did, and provided better blood pressure control during exercise, observed as a reduction in the variation of systolic blood pressure/MET (trandolapril: 10.7 1.9 mmHg/U vs 7.4 1.2 mmHg/U, p=0.02; captopril: 9.1 1.4 mmHg/U vs 11.4 2.5 mmHg/U, p=0.35), a reduction in peak diastolic blood pressure (trandolapril: 116.8 3.1 mmHg vs 108.1 2.5 mmHg, p=0.003; captopril: 118.2 3.1 mmHg vs 115.8 3.3 mmHg, p=0.35), and a reduction in the interruption of the tests due to excessive elevation in blood pressure (trandolapril: 50% vs 15%, p=0.009; captopril: 50% vs 45%, p=0.32). CONCLUSION: Monotherapy with trandolapril is more effective than that with captopril to control blood pressure during exercise in hypertensive patients. PMID- 12640508 TI - Mildly symptomatic chronic mitral regurgitation. Analysis of left ventricular systolic function and mitral regurgitation fraction under pharmacological influence. Echocardiographic study. AB - OBJECTIVE: To study echocardiographic parameters of left ventricular systolic function and valvar regurgitation under pharmacological influence in mildly symptomatic patients with chronic mitral regurgitation (MR). METHODS: We carried out a double-blind placebo controlled study in 12 patients with MR, mean aged 12.5 years old, who were randomized in 4 phases: A) digoxin; B) enalapril; C) digoxin + enalapril; D) placebo. The medication was administered for 30 days in each phase, and the following variables were analyzed: shortening and ejection fractions, wall stress index of left ventricle, left ventricular meridional end systolic wall stress, Doppler-derived mean rate of left ventricular pressure rise (mean dP/dt), stroke volume and MR jet area. The clinical variables analysed were heart rate and systemic arterial pressure. RESULTS: No significant variation was observed in the clinical variables analysed. The shortening and ejection fraction, the mean dP/dt and stroke volume significantly increased and the wall stress index of left ventricle, the meridional left ventricular end systolic wall stress and the mitral regurgitation jet area decreased in the phases with medication as compared with that in the placebo phase. CONCLUSION: The parameters of left ventricular systolic function improved significantly and the degree of MR decreased with the isolated administration of digoxin or enalapril in mildly symptomatic patients with chronic MR. The combination of the drugs, however, did not show better results. PMID- 12640509 TI - Comparison of the effect of two HMG CoA reductase inhibitors on LDL susceptibility to oxidation. AB - OBJECTIVE: To study the differences between fluvastatin and pravastatin regarding LDL susceptibility to oxidation, plasma levels of total cholesterol (TC), HDL-C, LDL-C and triglycerides (TG) in hypercholesterolemic patients with established coronary heart disease (CHD). METHODS: A double-blind randomized parallel study was conducted that included 41 hypercholesterolemic outpatients with CHD treated at the Instituto de Cardiologia do Rio Grande do Sul. The inclusion criteria were LDL-C above 100 mg/dL and triglycerides below 400 mg/dL based on 2 measures. After 4 weeks on a low cholesterol diet, those patients that fullfilled the inclusion criteria were randomized into 2 groups: the fluvastatin group (fluvastatin 40 mg/day) and the pravastatin group (pravastatin 20 mg/day), for 24 weeks of treatment. LDL susceptibility to oxidation was analyzed with copper induced production of conjugated dienes (Cu2+) and water-soluble free radical initiator azo-bis (2'-2'amidinopropanil) HCl (AAPH). Spectroscopy nuclear magnetic resonance was used for determination of lipids. RESULTS: After 24 weeks of drug therapy, fluvastatin and pravastatin significantly reduced LDL susceptibility to oxidation as demonstrated by the reduced rate of oxidation (azo and Cu) and by prolonged azo-induced lag time (azo lag). The TC, LDL-C, and TG reduced significantly and HDL-C increased significantly. No differences between the drugs were observed. CONCLUSION: In hypercholesterolemic patients with CHD, both fluvastatin and pravastatin reduced LDL susceptibility to oxidation. PMID- 12640510 TI - Impact of nutritional counseling in reducing serum cholesterol in public health service patients. AB - OBJECTIVE: To assess the impact of nutritional attention on the lipid profile and nutritional status of hypercholesterolemic patients attended in health centers of Belo Horizonte. METHODS: Using nutritional attendance patient record cards from two health units, the evolution of the lipid profile and the nutritional state (BMI) was monitored of 96 hypercholesterolemic patients who received diet. The patients were appraised at the following moments: initial (1st consultation), after 3 months (2nd consultation) and last consultation (variable for each patient). RESULTS: On the first attendance, 44,4% of the patients presented not only high total cholesterol and LDL-c, but also hypertriglyceridemia and 70.3% were overweight or obese, but most patients (75.6%) presented adequate HDL-c levels. There was significant reduction in the BMI, total cholesterol, LDL-c values (p < 0.01) and also in the triglyceride levels (p < 0.05) in the first three months, without alteration in the HDL-c levels. A significant reduction (p < 0.01) was observed in the frequency of individuals with high cholesterol (from 89.6% down to 47.9%), high and very high LDL-c (from 82.6% down to 45.7%), as well as high and very high triglyceride (from 43.6% down to 16.7%). The observed reduction in frequency of the low HDL-c was statistically meaningless. CONCLUSION: This study evidences the effect of the nutritional attention on lipid profile in hypercholesterolemic patients, reinforcing the need for a multiprofessional team to attend them at the public health services. PMID- 12640511 TI - Socioeconomic aspects of spousal concordance for hypertension, obesity, and smoking in a community of Rio de Janeiro, Brazil. AB - OBJECTIVE: The present study aims to evaluate the environmental role in the distribution of hypertension, obesity, and smoking and spousal concordance for the presence/absence of these 3 cardiovascular risk factors. METHODS: A cross sectional study was conducted in a community in Rio de Janeiro, Brazil. The households were randomly selected. Odds ratios were estimated to measure spousal concordance, across socioeconomic levels. RESULTS: Overall a significant aggregation of all 3 risk factors was present. The crude odds ratio for hypertension was 1.78 (95%CI=1.02-3.08); for obesity, it was 1.80 (95%CI=1.09 2.96); and for smoking, it was 3.40 (95% CI=2.07-5.61). The spousal concordance for hypertension decreased significantly (p<0.001) from the lower to the higher educational level. In the case of obesity and smoking, the opposite was observed, although p-values for the linear trend were 0.10 and 0.08, respectively. CONCLUSION: In lower socioeconomic levels, couples are more concordant for hypertension and discordant for smoking. For hypertension and smoking, education seems to be a discriminant stronger than income, but for obesity the 2 socioeconomic indicators seem to represent different aspects of the environmental determinants of risk factor distribution. PMID- 12640512 TI - Can patients with left main coronary artery disease wait for myocardial revascularization surgery? AB - OBJECTIVE: To assess the occurrence of cardiac events in patients diagnosed with left main coronary artery disease on diagnostic cardiac catheterization and waiting for myocardial revascularization surgery. METHODS: All patients diagnosed with left main coronary artery disease (stenosis > or = 50%) consecutively identified on diagnostic cardiac catheterization during an 8-month period were selected for the study. The group comprised 56 patients (40 males and 16 females) with a mean age of 61 10 years. The cardiac events included death, nonfatal acute myocardial infarction, acute left ventricular failure, unstable angina, and emergency surgery. RESULTS: While waiting for surgery, patients experienced the following cardiac events: 7 acute myocardial infarctions and 1 death. All events occurred within the first 60 days after the diagnostic cardiac catheterization. More patients, whose indication for diagnostic cardiac catheterization was unstable angina, experienced events as compared with those with other indications [p=0.03, relative risk (RR) = 5.25, 95% confidence interval = 1.47 - 18.7]. In the multivariate analysis of logistic regression, unstable angina was also the only factor that independently contributed to a greater number of events (p = 0.02, OR = 8.43, 95% CI =1.37 - 51.7). CONCLUSION: Unstable angina in patients with left main coronary artery disease acts as a high risk factor for cardiac events, emergency surgery being recommended in these cases. PMID- 12640513 TI - Right ventricular diastolic dysfunction in the postoperative period of tetralogy of Fallot. AB - OBJECTIVE: To assess right ventricular diastolic function in the intermediate postoperative period of repair of tetralogy of Fallot. METHODS: We carried out a case-control study with 60 patients divided into 2 groups as follows: 1) group I 30 patients who had undergone repair of tetralogy of Fallot and 2) group II - 30 healthy children. The 2 groups were paired for age, sex, and body surface. The flows in the pulmonary and tricuspid valves were analyzed with Doppler echocardiography. The presence of anterograde flow at the end of diastole in the pulmonary artery defined restrictive right ventricular physiology. Surgical, radiological, electrocardiographic, and echocardiographic variables were analized in the group I. RESULTS: The velocity of the A wave and the E/A ratio for the tricuspid valve showed significant differences between the groups. Cases with E/A < 1.30 predominated in inspiration (group I - 19/30, and group II - 5/30). The duration of the QRS complex on the electrocardiogram was significantly increased in patients with E/A <1.30. Nineteen (63.3%) patients had restrictive right ventricular physiology, which had a longer postoperative period, longer duration of the QRS complex, and a lower E/A ratio in inspiration. The surgical and radiological variables showed no statistical difference. CONCLUSION: Restrictive right ventricular physiology was detected on the intermediate follow-up of most patients undergoing repair of tetralogy of Fallot. The postoperative period and QRS duration were increased in patients with impairment in diastolic function. PMID- 12640514 TI - Value of the radiological study of the thorax for diagnosing left ventricular dysfunction in Chagas' disease. AB - OBJECTIVE: To determine the value of the radiological study of the thorax for diagnosing left ventricular dilation and left ventricular systolic dysfunction in patients with Chagas' disease. METHODS: A cross-sectional study of 166 consecutive patients with Chagas' disease and no other associated diseases. The patients underwent cardiac assessment with chest radiography and Doppler echocardiography. Sensitivity, specificity, and positive and negative predictive values of chest radiography were calculated to detect left ventricular dysfunction and the accuracy of the cardiothoracic ratio in the diagnosis of left ventricular dysfunction with the area below the ROC curve. The cardiothoracic ratio was correlated with the left ventricular ejection fraction and the left ventricular diastolic diameter. RESULTS: The abnormal chest radiogram had a sensitivity of 50%, specificity of 80.5%, and positive and negative predictive values of 51.2% and 79.8%, respectively, in the diagnosis of left ventricular dysfunction. The cardiothoracic ratio showed a weak correlation with left ventricular ejection fraction (r=-0.23) and left ventricular diastolic diameter (r=0.30). The area calculated under the ROC curve was 0.734. CONCLUSION: The radiological study of the thorax is not an accurate indicator of left ventricular dysfunction; its use as a screening method to initially approach the patient with Chagas' disease should be reevaluated. PMID- 12640515 TI - Tetralogy of Fallot and hypertrophic cardiomyopathy: a rare association. AB - Tetralogy of Fallot is known as the most common cyanotic congenital heart disease and has a prevalence of 10% of all congenital heart diseases. Although many other heart anomalies may coexist, the association of tetralogy of Fallot and hypertrophic cardiomyopathy is extremely rare. We report this association in a 15 month-old female, cyanotic since birth, in her first hospital admission for diagnosis and treatment of recurring cyanotic crises. In addition, a review of the literature and of the problems related to the treatment is provided. PMID- 12640516 TI - [Anatomoclinical correlation. Case 1/03 - male, 27 years-old, with reagent serology for Chagas disease and history of rheumatic fever for 11 years (Pontificia Universidade Catolica - Campinas, SP)]. PMID- 12640517 TI - [Clinico-radiographic correlation. Case 2/2003 - Instituto do Coracao do Hospital das Clinicas da FMUSP]. PMID- 12640518 TI - [Standardization of equipments and techniques for exams of ambulatory blood pressure mapping and home blood pressure monitoring]. PMID- 12640519 TI - Disruption of mitochondrial function in Candida albicans leads to reduced cellular ergosterol levels and elevated growth in the presence of amphotericin B. AB - A respiratory-deficient mutant of Candida albicans MEN was generated by culturing cells in medium supplemented with ethidium bromide at 37 degrees C for 5 days. The respiratory-deficient mutant (C. albicans MMU11) was incapable of growth on glycerol, had a reduced oxygen uptake rate and demonstrated an altered mitochondrial cytochrome profile. Respiratory-competent cybrids were formed by mitochondrial transfer following fusion of protoplasts with those of C. albicans ATCC 44990. Mutant MMU11 possessed lower levels of ergosterol than the parental isolates and the cybrids, and demonstrated a small but statistically significant increase in tolerance to amphotericin B. The results demonstrated that disruption of mitochondrial function in C. albicans increases the tolerance to amphotericin B, possibly mediated by a reduction in cellular ergosterol content. PMID- 12640520 TI - Exploring the Penicillium marneffei genome. AB - Penicillium marneffei is a dimorphic fungus that intracellularly infects the reticuloendothelial system of humans and bamboo rats. Endemic in Southeast Asia, it infects 10% of AIDS patients in this region. The absence of a sexual stage and the highly infectious nature of the mould-phase conidia have impaired studies on thermal dimorphic switching and host-microbe interactions. Genomic analysis, therefore, could provide crucial information. Pulsed-field gel electrophoresis of genomic DNA of P. marneffei revealed three or more chromosomes (5.0, 4.0, and 2.2 Mb). Telomeric fingerprinting revealed 6-12 bands, suggesting that there were chromosomes of similar sizes. The genome size of P. marneffei was hence about 17.8-26.2 Mb. G+C content of the genome is 48.8 mol%. Random exploration of the genome of P. marneffei yielded 2303 random sequence tags (RSTs), corresponding to 9% of the genome, with 11.7, 6.3, and 17.4% of the RSTs having sequence similarity to yeast-specific sequences, non-yeast fungus sequences, and both (common sequences), respectively. Analysis of the RSTs revealed genes for information transfer (ribosomal protein genes, tRNA synthetase subunits, translation initiation, and elongation factors), metabolism, and compartmentalization, including several multi-drug-resistance protein genes and homologues of fluconazole-resistance gene. Furthermore, the presence of genes encoding pheromone homologues and ankyrin repeat-containing proteins of other fungi and algae strongly suggests the presence of a sexual stage that presumably exists in the environment. PMID- 12640521 TI - Length of ICU stay for chronic obstructive pulmonary disease varies among large community hospitals. AB - OBJECTIVE: To determine whether differences exist among large community hospitals in length of Intensive Care Unit (ICU) stay, hospital stay or hospital mortality for patients admitted to ICU and whose most responsible diagnosis was chronic obstructive pulmonary disease (COPD). DESIGN: Retrospective cohort study. SETTING: All seven large community hospitals in British Columbia, Canada. PATIENTS. All 296 patients who were admitted to ICUs and whose most responsible diagnosis was COPD during the 3 fiscal years 1994-1997. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: After adjusting for age, gender, case-mix group, and co-morbidity, we found a significant difference in length of ICU stay for these patients among hospitals ( P <0.03). No differences were found in hospital mortality or length of hospital stay for the same patients among the same hospitals. CONCLUSIONS: There is significant variation in length of ICU stay for patients who are admitted to ICU and whose most responsible diagnosis is COPD, among large community hospitals. These small area variations may point to opportunities to improve efficiency of care. Further prospective, detailed data collection is required to validate these observations and to identify factors responsible for any differences found. PMID- 12640522 TI - Undifferentiated mesenchymal neoplasm of the esophagus in a child: case report and comparison with gastrointestinal stromal tumor. AB - Mesenchymal neoplasms of the esophagus are relatively uncommon in adults and exceedingly rare in children. Childhood tumors consist almost exclusively of smooth muscle tumors (leiomyomas). We report a case of an undifferentiated mesenchymal neoplasm occurring in the distal esophagus of a 15-year-old boy which is not a gastrointestinal stromal tumor. To our knowledge, this is the first reported case of such a neoplasm occurring in the esophagus of either an adult or child. PMID- 12640523 TI - Bacterial hemorrhagic enterocolitis. AB - Bacterial diarrhea can be classified into two clinical entities, noninflammatory diarrhea and inflammatory diarrhea syndromes. The latter type of diarrhea is characterized by bloody and puruloid mucus stool, and is often accompanied by fever, tenesmus, and severe abdominal pain. Pathogenic bacteria causing the inflammatory diarrhea syndrome include Salmonella, Vibrio, Shigella, enteroinvasive and enterohemorrhagic Escherichia coli, Campylobacter, Yersinia, Chlamydia, and Clostridium difficile. The pathologic changes in the inflammatory diarrhea syndrome range from a superficial exudative enterocolitis to a transmural enterocolitis with overt ulceration. This syndrome is also designated as bacterial hemorrhagic enterocolitis because of its usual manifestation by bloody diarrhea. The diagnostic approach needs information on the patient's age, travel history, epidemiological associations, sexual practice, and medical history, including usage of antibiotics. Bacterial information can be obtained by microscopic study, culture, and the identification of specific bacterial toxins. Flexible colonoscopy with biopsy is useful for the differentiation of bacterial hemorrhagic enterocolitis from idiopathic ulcerative colitis and ischemic colitis. Physicians should be familiar with the diagnostic modalities used to detect the specific pathogens causing hemorrhagic bacterial enterocolitis; namely, bacterial culture, serology, histology, and nucleic acid technologies. PMID- 12640524 TI - Anti-Saccharomyces cerevisiae antibodies in Japanese patients with inflammatory bowel disease: diagnostic accuracy and clinical value. AB - BACKGROUND: The diagnostic accuracy of the determination of anti- Saccharomyces cerevisiae antibodies (ASCA) and its clinical significance in Crohn's disease (CD) have been reported in Western countries, but scarcely reported in Japan, where the dietary habits of people differ markedly from those of Western countries. Th present study was undertaken to examine the diagnostic accuracy and clinical significance of ASCA determination in Japanese patients with CD or ulcerative colitis (UC). METHODS: Seventy-five serum samples obtained from 68 patients with CD, 34 serum samples obtained from 30 patients with UC, 35 serum samples from 35 patients with liver cirrhosis, and 31 serum samples from 31 healthy controls were examined. The optical density of each sample was measured by an enzyme-linked immunosorbent assay (ELISA) method to quantify ASCA IgA and IgG. RESULTS: The cutoff level, as determined by discriminant analysis of the data, was 0.1502 for ASCA IgA and 0.156 for ASCA IgG. Of the 68 patients with CD, 31 (45.6%) were ASCA-positive, and of the 30 patients with UC, 4 (13.3%) were ASCA-positive according to these cutoff levels. The sensitivity and specificity of ASCA determination for the differential diagnosis of CD from UC were 45.6% and 86.7%, respectively. When the relationship between ASCA and the clinical features of CD was analyzed, ASCA positivity was found to be correlated with duration of illness. CONCLUSIONS: The specificity of ASCA was high; however, the sensitivity was not. ASCA titers were generally low in the Japanese population examined. It would be desirable to determine cutoff levels for ASCA tailored to the Japanese people for the diagnosis of inflammatory bowel disease. PMID- 12640525 TI - A comparison of the effects of medium- and long-chain triglycerides on neutrophil stimulation in experimental ileitis. AB - BACKGROUND: In ileitis, the chain length of dietary fats affects inflammation, and medium-chain triglycerides (MCTs), but not long-chain triglycerides (LCTs), reduce intestinal damage. The mechanism of this effect has not been fully elucidated. In this work, we studied the effects of MCTs and LCTs on polymorphonuclear neutrophil (PMN) action in trinitrobenzene sulfonic acid (TNB) induced ileitis. METHODS: Twelve-week-old male Sprague-Dawley (SD) rats received TNB in the ileal lumen and were then fed MCTs or LCTs for 3 days. RESULTS: We detected no significant differences in the morphological damage between the MCT and the LCT groups. The content of interleukin (IL)-8, on the other hand, was significantly lower in the MCT group than in the LCT group, as was myeloperoxidase activity. The CD11b expression by PMNs was higher in the LCT group, but the difference was not of statistical significance. CONCLUSIONS: These findings suggested that proinflammatory activity was greater in the LCT group in comparison with the MCT group. PMID- 12640526 TI - Germinated barley foodstuff, a prebiotic product, ameliorates inflammation of colitis through modulation of the enteric environment. AB - BACKGROUND: Germinated barley foodstuff (GBF), which contains glutamine-rich protein and hemicellulose-rich fiber, exhibits therapeutic effects in ulcerative colitis; however, its mechanism is still under investigation. The aim of this study was to evaluate the anti-inflammatory effects of GBF on colitis in terms of the epithelial inflammatory response. METHODS: Mice with dextran sulfate sodium induced colitis were used. The effects of GBF on the colitis were evaluated by measuring the body weight; disease activity; mucosal damage (histology, mucosal inflammatory parameters, nuclear factor kappa B [NFkB] activation, and signal transducer and activator of transcription 3 [STAT3]); serum interleukin 6 (IL-6) level; cecal short-chain fatty acids (SCFAs); and bile acid contents. RESULTS: GBF significantly prevented disease activity and body weight loss after induction of colitis. Serum IL-6 level and mucosal STAT3 expression were also significantly attenuated, with a conspicuous reduction of mucosal damage; NFkB activity showed the same tendency. Cecal butyrate content was significantly higher and, interestingly, GBF mice had lower bile acid concentrations than the control group. CONCLUSIONS: GBF has the potential to reduce the epithelial inflammatory response by depressing STAT-3 expression and inhibiting NFkB binding activity. These effects may be brought about by an increase of butyrate production and adsorption of bile acids. PMID- 12640527 TI - SEN virus infection in patients with chronic liver disease and hepatocellular carcinoma in Thailand. AB - BACKGROUND: SEN virus (SENV) has been recently identified as a candidate agent of non-A-E hepatitis virus. However, the exact role of this novel virus in the pathogenesis of chronic liver disease, including chronic hepatitis and cirrhosis, and the development of hepatocellular carcinoma (HCC) remains to be established. METHODS: Using seminested polymerase chain reaction (PCR) amplification to detect SENV-D and SENV-H strains in serum, we investigated SENV infection in voluntary blood donors and in patients with chronic liver disease and HCC. RESULTS: SENV was detected in 5 of 100 blood donors (5%), in 15 of 60 patients with chronic liver disease (25%), and in 25 of 60 patients with HCC (42%). The prevalence of SENV in patients with HCC was higher than that in patients with chronic liver disease (P = 0.05) and in blood donors (P < 0.001). An age-specific prevalence of SENV was found at high levels among individuals aged 21-40 years, but was not detected among individuals in the lower age group. No differences between SENV infected and non-infected patients were demonstrated with respect to demographic data, assumed source of infection, biochemical abnormalities, and severity of chronic liver disease and HCC. Moreover, SENV infection had no apparent effect on the survival of patients with HCC. CONCLUSIONS: Our data suggest that SENV infection is frequent among patients with chronic liver disease and HCC. However, pathogenic effects associated with SENV infection in chronic liver disease and HCC need further investigation. PMID- 12640528 TI - Interleukin 6 concentration in ascitic fluid of cirrhotic patients: relationship with previous episodes of spontaneous bacterial peritonitis. AB - BACKGROUND: We aimed to determine whether inter-leukin (IL)-6 concentrations in plasma and ascitic fluid from cirrhotic patients with previous episodes of spontaneous bacterial peritonitis (SBP) differred from those with no previous episodes of SBP. METHODS: Cirrhotic patients with ascites were divided into two groups: G1, without previous episodes of SBP (n = 6), and G2, with previous episodes of SBP (n = 6). Patients with chronic heart failure (n = 5) with ascites, without hepatic diseases, were included as controls (G3). IL-6 concentration was determined in ascitic fluid and plasma by enzyme-linked immunosorbent assay (ELISA). A group of healthy subjects (n = 8) was used as a control for the blood IL-6 assay. RESULTS: IL-6 concentration in ascitic fluid was significantly higher in G1 (P < 0.05) than in G2 or G3. No significant difference in plasma IL-6 concentrations was observed between the groups or between each group and the healthy subjects. A significant positive linear correlation (P < 0.002) was observed between IL-6 concentration and total protein in ascitic fluid. CONCLUSIONS: A low concentration of IL-6 in ascitic fluid could predispose cirrhotic patients to SBP. PMID- 12640529 TI - Correlation between ultrasonographic and pathologic diagnoses of hepatitis B and C virus-related cirrhosis. AB - BACKGROUND. We aimed to evaluate the validity of ultrasonography (US) in the diagnosis of cirrhosis in patients with chronic hepatitis B virus (HBV) or C virus (HCV) infection. METHODS: A total of 210 patients, 67 with chronic HBV and 143 with HCV infection, were evaluated for the cirrhotic status of liver by both needle biopsy and US. According to the pathological findings, a fibrosis score 4 on the histology activity index was the gold standard for the diagnosis of cirrhosis. A US scoring system consisting of liver surface, parenchyma, vascular structure, and splenic size was used to describe the severity of hepatic parenchymal damage. RESULTS: Cirrhosis was found in 27 (40%) of the 67 HBV patients and in 51 (36%) of the 143 HCV patients pathologically. The mean fibrosis scores were 0.95, 1.24, 2.35, 2.95, 3.8 and 3.7 in patients with US scores of 4, 5, 6, 7, 8, and 9 or more, respectively. The US scores were significantly correlated with the hepatic fibrosis scores (P < 0.05). Based on the receiver operating characteristic (ROC) curve, a US score of 7 was the best cutoff point for the prediction of HBV-related cirrhosis, with sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of 77.8%, 92.5%, 87.5%, 86.0%, and 86.6%, respectively. In HCV-related cirrhosis, a US score of 6 provided results of 82.4%, 70.7%, 60.9%, 87.8%, and 74.8%, respectively. The specificity, positive predictive value, and accuracy were significantly higher in patients with HBV than in those with HCV infection (P = 0.012, P = 0.032, and P= 0.079, respectively). CONCLUSIONS: Cirrhosis can be predicted well by US, especially in patients with HBV infection. PMID- 12640530 TI - Efficacy of prolonged interferon therapy for patients with chronic hepatitis C with HCV-genotype 1b and high virus load. AB - BACKGROUND: In patients with hepatitis C virus (HCV)-genotype 1b and a high virus load, of more than 1 Meq/ml by the DNA probe assay, the clearance of HCV-RNA was achieved in only 10% with a 6-month interferon (IFN) course. We therefore assessed the efficacy of prolonged IFN therapy in patients with HCV-genotype 1b and a high virus load. METHODS: A total of 51 patients with HCV genotype 1b who were given 6 million units (MU) of natural IFN-alpha daily for 8 weeks followed by three-times-weekly treatment with natural IFN-alpha for 16 weeks, were enrolled in this trial. These 51 patients were randomly assigned to one of two schedule groups at the time of termination of the first IFN therapy. The 48-week group patients (n = 25) were given 6 MU of natural IFN-alpha by intramuscular injection three times weekly for 24 weeks, beginning within a week after the termination of the first IFN therapy. The 72-week-group patients (n = 26) were given 6 MU of IFN-alpha by intramuscular injection three times a week for 48 weeks, beginning within a week after the termination of the first IFN therapy. The therapeutic efficacy was evaluated 24 and 30 months after the initiation of the first IFN treatment. A virological response (VR) to IFN therapy was defined as the normalization of serum alanime amino transferase (ALT) level (ALT less, similar 50 IU) and HCV-RNA negativity at the two time points. Biochemical response (BR) was defined as the normalization of serum ALT, but positivity for HCV-RNA, assessed by commercial Amplicor HCV qualitative assays, at the two time points. RESULTS: The efficacy of IFN treatment was assessed in relation to the IFN administration schedule by intention-to-treat (ITT) analysis and per-protocol analysis. With respect to the IFN regimen, VR occurred in 16.6% (4/24) of the patients in the 48-week-group with additional IFN and in 20% (5/25) in the 72 week-group with additional IFN by ITT analysis. The BR rate was 33.3% (8/24) in the 48-week group and 48% (12/25) in the 72-week group. CONCLUSIONS: We found that prolonged IFN therapy could be a worthwhile treatment strategy for patients with HCV genotype 1b and a high serum virus load. PMID- 12640531 TI - Evaluation of gastric acid secretion in two patients (each aged over 90 years) with Helicobacter pylori-negative nonsteroidal anti-inflammatory drug-caused duodenal ulcers. AB - We treated two patients (each aged over 90 years) with Helicobacter pylori negative nonsteroidal anti-inflammatory drug (NSAID)-caused duodenal ulcers, and had the opportunity to determine gastric acidity by means of 24-h pH monitoring. Endoscopic and histological examination showed no remarkable atrophic change in the gastric mucosa. The gastric pH was low throughout the day and night, and the gastric pH > or = 3 holding time ratio during 24 h was 17.1% and 25.8%, respectively in the two patients, so it was considered that they had gastric acid secretion of the same level as that in normal subjects of the same age or that in the young without H. pylori infection. Because of the complication of reflux esophagitis with a hiatal hernia, rabeprazole sodium, one of the proton pump inhibitors (PPIs), was administered and both patients made excellent progress. In conclusion, gastric acid secretion in patients with H. pylori-negative NSAID caused duodenal ulcers is fully maintained even in the elderly, so PPIs may be the first choice of treatment. PMID- 12640532 TI - Rapidly growing early gastric cancer with microsatellite instability. AB - A 77-year-old man underwent gastrointestinal endoscopy that revealed a small flat elevated lesion in the gastric antrum. Endoscopically, we assessed the tumor as intramucosal cancer or adenoma, and biopsy specimens were diagnosed as showing borderline malignancy. At the second examination, performed 6 months later, the elevated lesion had grown, showing remarkable changes in its appearance, and a diagnosis of adenocarcinoma was confirmed pathologically. The patient underwent distal gastrectomy about 8 months after the first examination, and the tumor in the resected specimens showed a moderately differentiated adenocarcinoma invading through the entire submucosal layer, with metastasis to a regional lymph node. Microsatellite analysis of this early gastric cancer disclosed mutations in all six markers examined, indicating high-level microsatellite instability (MSI-H). The tumor also showed frameshift mutations of TGFbetaRII and BAXgenes, as well as hypermethylation of the hMLH1promoter. These results suggest that a deficiency of the mismatch repair system played a critical role in the progression of this cancer. PMID- 12640533 TI - Malignant pheochromocytoma associated with pseudo-obstruction of the colon. AB - Effective treatment has not yet been established for intestinal pseudo obstruction, a rare complication of malignant pheochromocytoma. We report the case of a 41-year-old man who presented with malignant pheochromocytoma associated with pseudo-obstruction of the colon. His serum catecholamine level was markedly elevated, and his large intestine was distended with gas and lodged stool. Laxatives and enemas were not effective in relieving his symptoms. We subsequently performed an ileostomy, after which the patient had good bowel movement and was able to resume oral food intake. PMID- 12640534 TI - Adult intussusception due to lymphangioma of the colon. AB - We present herein a case report of adult intussusception due to lymphangioma of the colon. On May 27, 2000, a 39-year-old woman with right lower abdominal pain was admitted to our hospital. Preoperative imaging studies, by ultrasonography, computed tomography (CT), and barium enema examination, showed right colon intussusception with a multilocular cystic tumor as a leading point. Emergency operation was performed. During the operation, normograde ileocecal intussusception with a 9 x 6-cm soft submucosal tumor of the cecum was recognized. Ileocecal resection was performed, and the patient's postoperative course was uneventful. Pathological diagnosis of the resected specimen was a cystic type lymphangioma of the cecum. Recently, lymphangioma of the colon has been diagnosed more frequently by colonoscopy and endoscopic ultrasonography. However, only a few cases of colon intussusception due to lymphangioma have been reported in the literature to date. Large lymphangioma of the colon is rare, but it should be taken into consideration that it is one of the organic lesions that causes adult intussusception. PMID- 12640535 TI - Gastrointestinal stromal tumor of the rectal mesentery. AB - We report a case of gastrointestinal stromal tumor (GIST) arising from the rectal mesentery. GIST of the large intestine is a rare tumor that accounts for only 0.1% of all colorectal cancers. The patient presented to our hospital with constipation and abdominal distension. Computed tomography (CT) revealed a huge mass in the pelvic cavity, and laparotomy disclosed diffuse peritoneal dissemination from the primary tumor. Radiochemotherapy was commenced, but the patient became too ill to complete it and died of the disease 2 months after surgery. A large high-grade tumor with diffuse dissemination was recognized as an indicator of poor survival in this patient. PMID- 12640536 TI - Intracystic hemorrhage of a simple liver cyst mimicking a biliary cystadenocarcinoma. AB - Simple liver cysts are rarely complicated by intracystic hemorrhage. We encountered a case of simple liver cyst that was morphologically similar to biliary cystadenocarcinoma, which was complicated by asymptomatic intracystic hemorrhage and successfully treated by right lobectomy. A large cystic lesion of the liver was detected in a 57-year-old woman during a mass screening health check. Abdominal ultrasonography (US) revealed that the cystic lesion, containing many hyperechoic papillary structures, occupied almost the entire region of the right hepatic lobe. In addition, a round mural nodule, measuring approximately 5 cm in diameter, was detected in the cystic wall. Abdominal computed tomography (CT) revealed that the inner part of the cystic lesion showed homogeneous low density, but CT did not show the round nodule detected by US. On T1-weighted sequence of magnetic resonance imaging (MRI), the lesion showed homogeneous high signals, together with a low-signal tumorous lesion in the cystic wall. T2 weighted sequence of MRI showed unhomogeneous high signals, together with high signals in the tumorous part. These findings did not exclude the possibility of a malignant cystic tumor, such as biliary cystadenocarcinoma. Therefore, right lobectomy was performed. Histological examinations of resected tissue specimens revealed that the lesion was a liver cyst containing a large amount of blood clot, and that the tumorous lesion detected by US and MRI was a large mass of blood clot which was partly liquefied. This case indicates the diagnostic importance of the morphological discordance between CT and US or MRI findings for liver cyst containing a large amount of blood clot. PMID- 12640537 TI - A case of Meckel's diverticulum adherent to the posterior abdominal wall: the efficacy of small-bowel radiography coupled with barium enema examination. PMID- 12640538 TI - Hepatocellular carcinoma complicating autoimmune hepatitis without either hepatitis C viral infection or corticosteroid therapy. PMID- 12640539 TI - Interaction between luminal bacteria and innate immunity. PMID- 12640540 TI - What features of the pathological morphology of the liver increase the accuracy of diagnostic imaging for cirrhosis? PMID- 12640541 TI - Indication of prolonged interferon therapy for chronic hepatitis C. PMID- 12640542 TI - Postoperative cell mediated immune response is better preserved after laparoscopic vs open colorectal resection in humans. AB - BACKGROUND: Animal studies have documented significantly better preserved postoperative cell-mediated immune function, as measured by serial delayed-type hypersensitivity (DTH) challenges, after laparoscopic-assisted than after open bowel resection. Similarly, in humans, the DTH responses after open cholecystectomy have been shown to be significantly smaller than preoperative responses; whereas after laparoscopic cholecystectomy, no significant change in DTH response has been noted. The purpose of this study was to assess cell mediated immune function via serial DTH skin testing in patients undergoing laparoscopic or open colectomy. METHODS: A total of 35 subjects underwent either laparoscopic (n = 18) or open colectomy (n = 17) in this prospective but not randomized study. Only patients who were judged to be immunoresponsive by virtue of having responded successfully to a preoperative DTH challenge were eligible for entry in the study. DTH challenges were carried out at three time points in all patients: preoperatively, immediately following surgery, and on the third postoperative day (POD 3). Responses were measured 48 h after each challenge and the area of induration calculated. There were no significant differences between the laparoscopic (LC) and open (OC) colorectal resection groups in regard to demographics, indications for surgery, or type of resection carried out. The percentage of patients transfused was similar in both groups (17%, LC; 12% OC; p = NS). In the LC group, all cases were completed without conversion using minimally invasive methods. There were no perioperative deaths, and the rate of postoperative complications was similar in both groups. The preoperative and postoperative DTH results were analyzed and compared within each surgical group using several methods. RESULTS: In regards to the OC group results, the median sum-total DTH responses for the day of surgery challenges (0.44 +/- 69 cm2) and the POD 3 challenges (0.72 +/- 3.37 cm2) were significantly smaller than the preoperative results (3.61 +/- 3.83 cm2, p <0.0005 vs op day and p <0.0003 vs POD 3 results). When the LC group results were similarly analyzed, no significant difference in DTH response was noted between the pre- and the postoperative challenge results. Additionally, when the median percent change from baseline was calculated and considered for the OC group's DTH results, both postoperative challenge time points demonstrated significantly decreased responses when compared to their preoperative results (vs day of surgery, p <0.007; vs POD 3, p <0.006). Similar analysis of the LC group's results yielded nonsignificant differences between the pre- and postoperative responses. Lastly, when the LC and the OC groups median percent change from baseline results were directly compared for each of the postoperative challenges, a significant difference was noted for the POD 0 challenge (LC, -21%; OC 88%; p <0.004) but not for the POD 3 challenge. CONCLUSIONS: The postoperative DTH responses of the open surgery patients were significantly smaller than their preoperative responses. This was not the case for the laparoscopic group (a combination of fully laparoscopic and laparoscopic assisted resections). When the open and laparoscopic groups results are directly compared, regarding the results of the day of surgery DTH challenges, the LC groups median percent change from baseline was significantly less than that observed in the OC group. These results imply that open colorectal resection is associated with a significant suppression of cell-mediated immune response postoperatively, whereas in this study laparoscopic colorectal resection was not. Further human studies are needed to verify these findings and to determine the clinical significance, if any, of this temporary difference in immune function following colon resection. PMID- 12640543 TI - Surgical smoke: a review of the literature. Is this just a lot of hot air? AB - Surgical smoke is omnipresent in the day-to-day life of the surgeon and other medical personnel who work in the operating room. In addition, patients are also exposed, especially and uniquely so in laparoscopic cases where smoke is created and trapped in a closed and absorptive space. Surgical smoke has typically been produced by electrocautery but is now ever more present in a new form with the burgeoning use of the laser and the harmonic scalpel. Several cases of transmission of human papillomavirus (HPV) from patient to treating professional via laser smoke have alerted us to the reality that surgical smoke in certain situations is far form benign. However, surgeons rarely take measures to protect themselves, their co-coworkers and patients from surgical smoke. Should we and, if so, how do we differentiate between different types of smoke and should we move toward increasing our efforts to protect ourselves, our co-workers, and patients from it? This article attempts to sort through the available data and draw some reasonable conclusions regarding surgical smoke. In general, surgical smoke is a biohazard and cannot be ignored. At a minimum, surgical smoke is a toxin similar to cigarette smoke. However, other dangers exist. This is especially true in specific circumstances such as when tissue infected with dangerous viruses is aerosolized by lasers. In addition, smoke generated by the harmonic scalpel, being a relatively cold vapor similar to laser smoke, should be further investigated for its potential ill effects and until then, looked upon with reasonable caution. Although not a high-priority in most surgical cases, surgeons should support efforts to minimize OR personnel, patients, and their own exposure to surgical smoke. PMID- 12640544 TI - Fetal DNA in maternal serum: does it persist after pregnancy? AB - Fetal DNA and cells present in maternal blood have previously been used for non invasive prenatal diagnosis. However, some fetal cells can persist in maternal blood after a previous pregnancy. Fetal rhesus status and sex determination have been performed by using amplification by real-time polymerase chain reaction (PCR) of fetal DNA sequences present in maternal circulation; no false-positive results related to persistent fetal DNA from a previous pregnancy have been reported. This idea has recently been challenged. An SRY real-time PCR assay was performed on the serum of 67 pregnant women carrying a female fetus but having previously given birth to at least one boy and on the serum of 30 healthy non pregnant women with a past male pregnancy. In all cases, serum was negative for the SRY gene. These data suggest that fetal DNA from a previous pregnancy cannot be detected in maternal serum, even by using a highly sensitive technique. Therefore, non-invasive prenatal diagnosis by fetal sex determination for women at risk of producing children with X-linked disorders, and fetal RHD genotyping is reliable and secure as previously demonstrated. PMID- 12640545 TI - [Cataract prevention. Therapeutic approaches and critical review of current status]. AB - Research on the pathophysiology of the lens already in the early days of the last century led to first attempts to clinically influence cataract development with vitamins. More detailed investigation of lens aging and its interaction with internal and external cataract risk factors led to two different therapeutic strategies: (1) compounds or mixtures expected to slow down the aging processes in the lens (food additives) and (2) compounds to reduce or even arrest the effect of a specific harmful factor. Various mixtures expected to influence aging processes were even developed into approved OTC drugs although their effectiveness was never demonstrated. Among those compounds designed to act on a specific pathomechanism, mainly aldose reductase inhibitors were designed and successfully tested in animal studies. None of these,however, could be developed into a market-approved drug. Larger controlled clinical studies have been performed with various compositions of food additives, but also with nonsteroidal anti-inflammatory drugs such as salicylic acid and ibuprofen. None of the clinical trials,however, evidenced any convincing anti-cataract effect of the compounds or mixtures tested such that a successful anti-cataract drug still remains to be developed. PMID- 12640546 TI - [Antioxidant micronutrients and cataract. Review and comparison of the AREDS and REACT cataract studies]. AB - Age-related cataract remains the major cause of preventable blindness throughout the world. It has long been realized that one of the important etiological factors for this disease is oxidative and in particular photooxidative damage to the lens. Therefore, the antioxidant micronutrients, vitamins C and E and the carotenoids, in particular beta-carotene, have been discussed as factors that could reduce the risk for this disease. The present article reviews what is known about the transport of these substances to the lens, their accumulation, and their concentrations in the lens. Furthermore, the available epidemiological literature is briefly mentioned, but more emphasis has been placed on a description and discussion of major clinical intervention studies. Finally, the design and results of two of those trials using antioxidant micronutrients, the Age-Related Eye Disease Study (AREDS) and the Roche European American Cataract Trial (REACT), are compared. The AREDS trial did show a positive effect only for age-related macular degeneration but not for cataract, while the REACT trial demonstrated a small but statistically significant deceleration of cataract progression. The techniques for following the course of a cataract in the REACT study were more sensitive to subtle changes than those used in the AREDS study, and this may have been one important factor accounting for the differences. The authors' detailed comparison of these studies, however, suggests that even more important may have been the fact that in the REACT study intervention started earlier in the disease process,with higher doses of vitamins C and E and beta carotene and consequently with larger plasma concentrations of these antioxidant micronutrients. The REACT trial results support the early complementation of a diversified diet with supplements containing vitamins C and E and beta-carotene as well as other carotenoids. The authors also believe that it is reasonable to include these micronutrients in the therapeutic armamentarium of general ophthalmological practice. PMID- 12640548 TI - [Chorioretinal anastomosis as unfavourable prognostic factor during photodynamic therapy]. AB - BACKGROUND: The aim of this study was the documentation of chorioretinal anastomosis in neovascular age-related macular degeneration (AMD) and correlation with functional and angiographic results following photodynamic therapy (PDT). METHOD: A total of 100 patients presenting with neovascular AMD and indication for PDT based on the presence of predominantly classic choroidal neovascularization (CNV), underwent ophthalmoscopic and angiographic screening for chorioretinal anastomosis. Conventional PDT using verteporfin was performed according to the recommended standard procedure. The pre- and post-treatment status at 3 and 6 months post-PDT in all patients and at 1 week in selected patients was documented with respect to the central visual acuity test (ETDRS), ophthalmoscopy as well as fluorescein (FA) and indocyanine green angiography (ICGA). RESULTS: A primary chorioretinal anastomosis was found in 6% ( n=12) of all eyes with CNV and classic PDT indication. Mean visual loss within the first 6 months after therapy was 3 lines indicating lack of visual stabilization according to the PDT study criteria.Furthermore, an increase in visual acuity could not be documented in any case. Angiography demonstrated continuous progression of CNV size although PDT had been uneventful. The characteristic initial occlusion of the CNV with homogeneous hypofluorescence was absent angiographically at 1 week post-PDT. The anastomosis was detected by ICGA in all eyes and by ophthalmoscopy or optical coherence tomography (OCT) in most eyes. DISCUSSION: Chorioretinal anastomosis is not a rare finding associated with predominantly classic CNV. The presence of anastomosis appears to be an unfavourable prognostic factor during PDT. If a chorioretinal shunt is suspected evaluation by ICGA and/or OCT should be performed and the indication for PDT should be rediscussed. PMID- 12640547 TI - [Is a cataract avoidable? Current status with special emphasis on the pathophysiology of oxidative lens damage, nutritional factors, and the ARED study]. AB - The benefit of long-term nutrient intake to reduce the risk of age-related ocular disease such as cataract or macular degeneration is subject to controversy. Conclusions about the benefits and risks of antioxidant supplements can be expected after reviewing the current literature concerning oxidative-induced lens damage and nutritional effects. Identification of influenceable risk factors for senile cataracts could achieve immense economical relevance. In contrast to former longitudinal epidemiological studies, the AREDS report failed to verify protective properties of highly concentrated vitamin supplements on cataract formation. Although there are enough epidemiological indications for reducing the risk of cataracts by the intake of antioxidants, a general recommendation for the use of supplements is untimely or even wrong until stringent evidence of efficacy is provided. The usefulness of cataract prevention is discussed. PMID- 12640549 TI - [Experience with the artificial iris diaphragm in hypotonic eyes]. AB - BACKGROUND: As a result of a partial or complete loss of the natural iris diaphragm,longstanding silicone oil tamponade frequently causes keratopathy. An artificial iris diaphragm can avoid such a complication. In hypotony, with insufficient circulation of aqueous humor, the "closed" artificial iris diaphragm is used. METHODS: In this retrospective study 41 patients (41 eyes) were reviewed. Each patient had a single aphakic eye, in which a closed iris diaphragm was implanted after silicone oil surgery. The underlying diagnosis included in these 41 cases included trauma (22 eyes, 54%), retinal detachment due to proliferative vitreoretinopathy (12 eyes, 29%), severe uveitis (4 eyes, 10%), and proliferative diabetic retinopathy (3 eyes, 7%). The mean follow-up time was 12 month. RESULTS: In 25 eyes (61%) silicone oil was retained behind the diaphragm. In 14 eyes (34%), silicone oil prolapsed in the anterior chamber. In 2 eyes (5%) the silicone oil could not been assessed due to a corneal opacity.A deteriorated corneal situation after implantation of the diaphragm was observed in 11 eyes (27%). As a longstanding complication permanent hypotony (< or = 5 mmHg) developed in 29 eyes (71%), fibrous reaction in 13 eyes (32%) within fibrotic membranes (9 eyes, 22%). The visual acuity remained stable in 39 eyes (95%), improved in none, and deteriorated in 2 eyes (5%). CONCLUSION: Despite all complications, the artificial iris diaphragm represents an important progress in the salvage of severely traumatized eyes though persistent hypotony remains in 29 eyes (71%). Contact of silicone oil with the endothelium was avoided in 25 eyes (61%). There was no phthisis bulbi or enucleation. Improvement of anterior posterior separation is necessary. PMID- 12640550 TI - [Amniotic membrane transplantation improves herpetic keratitis by local and not by systemic effects]. AB - PURPOSE: The immune mediated HSV-1 stromal keratitis (HSK) rapidly improves after amniotic membrane transplantation (AMT). This study investigated whether AMT modulates the T cell response and whether the anti-inflammatory action of AMT is due to local or systemic effects. METHODS: Corneas of BALB/c mice were infected with 10(5) (PFU) of HSV-1. Animals with ulcerating keratitis on day 14 post infection were divided into 4 groups: group 1 ( n=12): right eye AMT;group 2 ( n=12): right eye tarsorrhaphy; group 3 ( n=8): right eye tarsorrhaphy, left eye AMT;group 4 ( n=8): both eyes tarsorrhaphy. The mice were examined for clinical signs of HSV keratitis after 2 days. Corneal sections were studied histologically and the inflammatory cell infiltration was studied by immunohistochemical staining. DTH response and the HSV-specific 3H-thymidin-uptake were compared between the groups. RESULTS: Compared to group 2, ulceration and stromal inflammation was profoundly improved in group 1 ( p<0.01). The corneas in the AMT mice had fewer inflammatory cells, CD3+,CD4+ and CD8+ cells than the control mice ( p<0.01). There were no significant differences between groups 1 and 2 with respect to the delayed type hypersensitivity reaction (DTH response) and the HSV specific 3H-thymidin uptake. AMT or tarsorrhaphy on the left eyes in groups 3 and 4 had no influence on the course of keratitis or the T cell response. CONCLUSIONS: Ulcerating herpetic keratitis markedly improves after AMT. Our observations indicate that this is caused predominantly by local and not by systemic AMT-related effects. PMID- 12640551 TI - [Anthrax--an old disease becomes topical: cutaneous anthrax and the eye]. AB - BACKGROUND: Anthrax disease and its eye manifestations were rare in the last 100 years, but the threat of terrorist actions has revived its topicality. MATERIALS AND METHODS: After an introductory historical review, the pathogenesis of this disease with regard to the virulence of Bacillus anthracis is reported. On the basis of photos displaying the course of the disease, the symptoms particularly of the cutaneous form of this disease as well as the diagnostic possibilities are described. RESULTS AND CONCLUSIONS: The current status of therapy and research for more effective treatment is discussed, with particular emphasis on the development of new substances with antitoxin properties and better vaccines. Bacillus anthracis is once again an actual threat, and therefore it is necessary for doctors to familiarize themselves with the current knowledge of this infection. PMID- 12640552 TI - [Corneal complications after hematopoietic stem cell transplantation]. AB - AIM: To evaluate the incidence and clinical course of corneal complications in patients with severe dry eye syndrome after hematopoietic stem cell transplantation (HSCT). PATIENTS AND METHODS: 50 consecutive patients (aged 9-65 years; average=42+/-11 years) with Sjogren-like syndrome after hematopoietic stem cell transplantation were examined. In order to assess the severity of the dry eye syndrome, the frequency of blinking and break-up time were determined and Schirmer-I, fluorescein,and rose bengal tests were carried out. Biopsy and histological examination were performed in cases with suspected conjunctival graft-versus-host reaction. RESULTS: Patients with Sjogren-like dry eye syndrome were referred within an average of 13 months after HSCT (SD+/-20 months).The follow-up was up to 83 months (mean: 10.2 months,SD+/-14.97). Of the 50 patients 15 developed severe corneal complications with significant loss of vision which resulted in enucleation of the eye in 1 patient. Of these 15 patients 4 had a viral and 2 a bacterial keratitis,7 had trophic corneal thinning and sterile ulcers. Two patients had limbal stem cell insufficiency or pseudomembranous conjunctivitis, both due to conjunctival graft-versus-host reaction. Another patient developed a toxic keratopathy, probably induced by cyclosporin A eye drops. Of 50 patients 35 presented with signs of an inactive or active conjunctival graft-versus-host reaction which could be proven histologically in 26 patients. DISCUSSION: Patients with Sjogren-like dry eye syndrome after hematopoietic stem cell transplantation are at high risk to develop corneal complications.These complications may result from an aggressive extension of the graft-versus-host reaction towards the conjunctiva and/or the lacrimal gland and seem to occur more often during the period of reduction of systemic immunosuppressive therapy.Furthermore, infectious as well as trophic or toxic corneal complications may be supported by local immunosuppressive therapy. We suggest frequent ophthalmological checks of patients receiving hematopoietic stem cell transplantation who need local immunosuppressive therapy or are in the phase of reduction of systemic immunosuppressive therapy PMID- 12640553 TI - [Combined deep sclerectomy and trabeculotomy in congenital glaucoma with complications]. AB - BACKGROUND: Lately there has been growing interest in deep sclerectomy as a therapeutic approach for the management of congenital glaucoma.In theory, its pressure-lowering effect is independent of external filtration, which minimizes the risk of failure due to conjunctival or episcleral scarring. In patients who have failed to benefit from previous filtering procedures and have refractory congenital glaucoma,deep sclerectomy could become an alternative. PATIENTS AND METHODS: A 3-year-old girl with refractory congenital glaucoma underwent deep sclerectomy in her left eye. Intraoperatively, percolation through the trabeculodescemetic window was weak, so that the procedure was supplemented by a trabeculotomy. RESULTS: By now, throughout a follow-up period of 20 months, stabilisation of disc cupping and ocular axial length has been achieved without additional glaucoma surgery or medications. CONCLUSIONS: In patients with congenital glaucoma,the combination of deep sclerectomy with trabeculotomy may provide an additional therapeutic approach whenever the prospect of success appears to be limited with deep sclerectomy alone. PMID- 12640554 TI - [Pigmented retinopathy as a presenting sign of mitochondrial encephalomyopathy without external ophthalmoplegia]. AB - BACKGROUND: Mitochondrial encephalomyopathies result from deletions in the nuclear or mitochondrial (mt) DNA. Deletions in the mtDNA are often sporadic. Mitochondriopathies are commonly associated with chronic progessive external ophthalmoplegia (CPEO). Here we describe a patient with a structural mtDNA aberration whose presenting sign was impaired visual acuity in the presence of a pigmented retinopathy but lack of impaired ocular motility. PATIENT: A 7-year-old girl presented with impaired visual acuity (0.4 OD and 0.5 OS), coarse hyperpigmentation of the posterior pole and diffuse hyperpigmentation with irregular depigmentation in the periphery. Scotopic and photopic as well as multifocal ERG were abnormal. Further symptoms included an incomplete inner ear deafness, ataxia, lapses of coordination and an intention tremor. Compared with her twin sister, the patient's speech was less modulated and slower. MRI scanning disclosed symmetric changes of density in the basal ganglia and nucleus dentatus as well as in the brainstem. ECG yielded no evidence of an AV-node block. Molecular biological analysis showed a structural rearrangement of the mtDNA. CONCLUSIONS: Mitochondrial encephalomyopathies in early ages may present with pronounced retinal changes in the absence of external ophthalmoplegia.Therefore, it appears prudent to include a neuropediatric evaluation as well as a mutation screening of the mtDNA in the evaluation of pediatric patients with diffuse non specific pigmented retinopathies. PMID- 12640555 TI - [Eyelid eczema. A therapeutic challenge]. PMID- 12640559 TI - A healthy hemophilic patient without arthropathy: from concept to clinical reality. AB - Hemophilia is characterized by joint hemorrhage with subsequent development of arthropathy. During the past 40 to 50 years, factor VIII concentrates have been developed that enable effective treatment of bleeding. However, the development of dosing, including the implementation of prophylaxis, has been hampered by concentrate transmission of blood-borne diseases. Since the advent of viral reducing procedures that were applied to plasma-derived products and the implementation of recombinant DNA technology into the manufacturing procedure, concentrates can be considered as safe today, and hemophilic care can be more focused on the treatment modalities as such. Thus, prophylaxis is being improved and made available to more patients. Studies are in progress in order to give more scientific evidence behind its use. Magnetic resonance imaging (MRI) seems promising in scaling early arthropathy and may be a tool in the future, superior to plain X-ray and orthopedic examination, to follow patients on prophylaxis, especially children. The development of inhibitors is the most severe threat to the health of hemophiliacs today and in the future. Much has been done to elucidate genetic factors predisposing for inhibitors, and the reactivity of inhibitors with the factor VIII molecule has also been investigated. More has to be done, however, and there are still many open issues regarding immune tolerance treatment and treatment of acute bleeding in inhibitor patients. Hemophilic care has come far in order to restore health for the hemophilic population. The treatment is effective in the prevention of joint disease and the inhibitor problem is handled better now. Side effects have been reduced to a minimum. In addition, carrier and prenatal diagnosis add important contributions to improve life for the patient and his family. The next important step for the future is gene therapy, and early clinical studies are already in progress. PMID- 12640557 TI - Antisense imaging of epidermal growth factor-induced p21(WAF-1/CIP-1) gene expression in MDA-MB-468 human breast cancer xenografts. AB - Molecular imaging of the expression of key genes which determine the response to DNA damage following cancer treatment may predict the effectiveness of a particular treatment strategy. A prominent early response gene for DNA damage is the gene encoding p21(WAF-1/CIP-1), a cyclin-dependent kinase inhibitor that regulates progression through the cell cycle. In this study, we explored the feasibility of imaging p21(WAF-1/CIP-1) gene expression at the mRNA level using an 18-mer phosphorothioated antisense oligodeoxynucleotide (ODN) labeled with (111)In. The known induction of the p21(WAF-1/CIP-1) gene in MDA-MB-468 human breast cancer cells following exposure to epidermal growth factor (EGF) was used as an experimental tool. Treatment of MDA-MB-468 cells in vitro with EGF (20 n M) increased the ratio of p21(WAF-1/CIP-1) mRNA/beta-actin mRNA threefold within 2 h as measured by the reverse transcription polymerase chain reaction (RT-PCR). A concentration-dependent inhibition of EGF-induced p21(WAF-1/CIP-1) protein expression was achieved in MDA-MB-468 cells by treatment with antisense ODNs with up to a tenfold decrease observed at 1 microM. There was a fourfold lower inhibition of p21(WAF-1/CIP-1) protein expression by control sense or random sequence ODNs. Intratumoral injections of EGF (15 microg/dayx3 days) were employed to induce p21(WAF-1/CIP-1) gene expression in MDA-MB-468 xenografts implanted subcutaneously into athymic mice. RT-PCR of explanted tumors showed a threefold increased level of p21(WAF-1/CIP-1) mRNA compared with normal saline treated tumors. Successful imaging of EGF-induced p21(WAF-1/CIP-1) gene expression in MDA-MB-468 xenografts was achieved at 48 h post injection of (111)In-labeled antisense ODNs (3.7 MBq; 2 microg). Tumors displaying basal levels of p21(WAF-1/CIP-1) gene expression in the absence of EGF treatment could not be visualized. Biodistribution studies showed a significantly higher tumor accumulation of (111)In-labeled antisense ODNs in the presence of EGF induction of the p21(WAF-1/CIP-1) gene (0.32%+/-0.06% injected dose/g) compared with normal saline-treated control mice (0.11%+/-0.07% injected dose/g). The tumor/blood ratio for antisense ODNs in the presence of EGF induction of the p21(WAF-1/CIP-1) gene (4.87+/-0.87) was also significantly higher than for control random sequence ODNs (2.14+/-0.69) or for mice receiving antisense ODNs but not treated with EGF (2.07+/-0.37). We conclude that antisense imaging of upregulated p21(WAF-1/CIP-1) gene expression is feasible and could represent a promising new molecular imaging strategy for monitoring tumor response in cancer patients. To our knowledge, this study also describes the first report of molecular imaging of the upregulated expression of a downstream gene target of the EGFR, a transmembrane tyrosine kinase receptor. PMID- 12640556 TI - Measuring [(18)F]FDG uptake in breast cancer during chemotherapy: comparison of analytical methods. AB - Over the years several analytical methods have been proposed for the measurement of glucose metabolism using fluorine-18 fluorodeoxyglucose ([(18)F]FDG) and positron emission tomography (PET). The purpose of this study was to evaluate which of these (often simplified) methods could potentially be used for clinical response monitoring studies in breast cancer. Prior to chemotherapy, dynamic [(18)F]FDG scans were performed in 20 women with locally advanced ( n=10) or metastasised ( n=10) breast cancer. Additional PET scans were acquired after 8 days ( n=8), and after one, three and six courses of chemotherapy ( n=18, 10 and 6, respectively). Non-linear regression (NLR) with the standard two tissue compartment model was used as the gold standard for measurement of [(18)F]FDG uptake and was compared with the following methods: Patlak graphical analysis, simplified kinetic method (SKM), SUV-based net influx constant ("Sadato" method), standard uptake value [normalised for weight, lean body mass (LBM) and body surface area (BSA), with and without corrections for glucose (g)], tumour to non tumour ratio (TNT), 6P model and total lesion evaluation (TLE). Correlation coefficients between each analytical method and NLR were calculated using multilevel analysis. In addition, for the most promising methods (Patlak, SKM, SUV(LBMg) and SUV(BSAg)) it was explored whether correlation with NLR changed with different time points after the start of therapy. Three methods showed excellent correlation ( r>0.95) with NLR for the baseline scan: Patlak10-60 and Patlak10-45 ( r=0.98 and 0.97, respectively), SKM40-60 ( r=0.96) and SUV(LBMg) ( r=0.96). Good correlation was found between NLR and SUV-based net influx constant, TLE and SUV(BSAg) (0.90< r<0.95). The 6P model and TNT had the lowest correlation ( r 1% and those that are guided by clinical bleeding patterns of individual patients. In both types of strategies, treatment should be started early, before the start of joint damage. However, clotting factor consumption in the first group of strategies is expected to be higher, whereas bleeding frequencies are expected to be slightly lower. The differences in long-term outcome between both strategies may only become apparent after more than two decades. In addition, some patients may be able to stop taking prophylaxis in adulthood while maintaining a low bleeding frequency. In conclusion, although the optimal prophylactic regimen is still under debate, it is well established that prophylaxis reduces or even prevents joint bleeds and hemophilic arthropathy. Therefore, prophylaxis should be offered to all children with severe hemophilia, at least until adulthood. PMID- 12640566 TI - Orthopedic outcome studies and cost issues. AB - Standard therapy for years was on-demand therapy. Readers will learn of results of clinical experience in many different countries with a prophylactic approach to therapy as well as ongoing experimental designs to define optimal therapy. The costs of current prophylaxis regimens will be discussed. PMID- 12640567 TI - Immunobiology of inhibitor development in hemophilia A. AB - After treatment with factor (F) VIII concentrate a significant number of patients with hemophilia A develop inhibitory antibodies that neutralize FVIII. Epitope mapping revealed that antibodies bind to selected regions within the A2, A3, and C2 domains of FVIII. Anti-A2 and anti-A3 antibodies interfere with assembly of FVIIIa with FIXa, whereas anti-C2 antibodies impede the interaction of FVIII with phospholipids. The immunologic mechanisms underlying inhibitor development in hemophilia A have not been fully elucidated. FVIII is recognized by the immune system as a foreign antigenic substance that evokes the T cell-dependent formation of high-affinity antibodies. Clonal analysis of B cell responses in hemophilia A patients has given further insight into the epitope specificity and molecular characteristics of FVIII inhibitors. Costimulatory blockade of FVIII reactive T cells in a mouse model for hemophilia A has suggested new approaches for treatment of inhibitor patients. In this article, recent studies on the immunobiology of FVIII inhibitors are summarized and discussed with reference to their potential impact on treatment and prevention of immune responses in patients with hemophilia. PMID- 12640569 TI - Treatment of the bleeding inhibitor patient. AB - The development of inhibitory antibodies to factor (F) VIII and FIX continues to be a major challenge in the treatment of patients with hemophilia. In patients with low-responding inhibitors, it is usually possible to saturate the inhibitor with the deficient factor and to achieve hemostasis, but in patients with high responding inhibitors, two major tasks have to be considered. One is how to treat the acute bleedings and the other is how to permanently eliminate the immune response, in other words, to induce tolerance. There are several hemostatic agents available for bleeding patients with high-responding inhibitors. Nonactivated and activated prothrombin complex concentrates (PCCs) have been used for almost 30 years, and since the beginning of the 1980s, porcine FVIII has also been used. In more recent years, recombinant FVIIa has been added to the therapeutic armamentarium and has been shown to control hemostasis in most patients. Immunoadsorption may temporarily reduce the inhibitor, enabling replacement therapy for several days. Available data on these alternative regimens will be discussed with a focus on the mechanisms of action, pharmacokinetics, safety, monitoring, and clinical experience. PMID- 12640568 TI - Immune tolerance induction in hemophilia A: a review. AB - In this article, a comparative analysis of the data stemming from the studies conducted in the field of immune tolerance treatment (ITT) of hemophilia A was attempted. Comparisons are difficult because previous studies differ in many respects, including the dosage of factor (F) VIII, the number of FVIII administrations per day, the association with immunosuppressive drugs (prednisone, cyclophosphamide), and, most importantly, the definition of success in terms of the reacquisition of tolerance. However, a number of variables consistently influenced outcome: the inhibitor titer, either the maximum one or the one assayed before immune tolerance (IT) start and age. As to the FVIII dose, results are contrasting, even though the overall impression is that high dosages are associated with higher success rates. Treatment duration analyses appear to be fairly consistent in all the studies, with 1 year as the crucial time period useful in predicting outcome. Preliminary data suggest that concentrates containing von Willebrand factor may increase the success rate and allow patients refractory to IT procedures carried out with ultrapure or recombinant concentrates to become tolerant in the end. These data need to be formally confirmed in trials that compare the different brands. In the studies published so far, a common language and widely accepted definitions of the variables have emerged, which are important in setting up the controlled trials and improving the comparison among published data. PMID- 12640570 TI - Management of musculoskeletal complications of hemophilia. AB - Prophylactic treatment from ages 2 to 18 years could prevent the development of hemophilic arthropathy if the concentration of the patient's deficient factor is kept from falling below 1% of normal. Early treatment is of paramount importance because the immature skeleton is very sensitive to the complications of hemophilia. Major hemarthrosis and chronic hemophilic synovitis should be treated aggressively to prevent hemophilic arthropathy. When advanced hemophilic arthropathy is present with severe disability, the aim should be to restore function while minimizing the risk to the patient. Joint debridement is an effective method to achieve this goal, especially around the elbow or ankle, and can be considered an alternative to knee replacement in the younger age groups. Proximal tibial valgus osteotomy is a reliable treatment method for painful genu varum of the mobile hemophilic knee. Supramalleolar tibial varus osteotomy is an attractive alternative to the more commonly used surgical option of arthrodesis. Finally, joint replacement can usually be relied on to restore both mobility and function in a diseased joint. The potential benefits of joint replacement must always be weighed against the long-term sequelae, however, especially loosening and consequent revision surgery. PMID- 12640571 TI - Radionuclide synovectomy (radiosynoviorthesis) in hemophilia: a very efficient and single procedure. AB - Radionuclide synovectomy, so-called radioactive synoviorthesis, is easy to perform and a not very expensive procedure (for example, 150 US dollars for an adult knee). Moreover, it is very efficient in diminishing the frequency and intensity of recurrent hemarthrosis related to chronic hypertrophic hemophilic synovitis. It should be performed as soon as possible in order to try to halt the long-term damage that intra-articular blood will produce in the involved joint. Personal experience and the general recommendation among orthopedic surgeons and hematologists is that when three early consecutive synoviortheses (repeated every 3 months) fail to halt synovitis, a surgical synovectomy (open or by arthroscopy) should be immediately considered. PMID- 12640572 TI - Desmopressin in mild hemophilia A: indications, limitations, efficacy, and safety. AB - Replacement therapy with blood products has long been the only available therapeutic option for patients with bleeding disorders. Plasma-derived cryoprecipitate and factor (F) VIII concentrates, which have been used for hemophilia A patients, involve the risk of transmitting blood-borne diseases. Both plasma-derived and recombinant FVIII concentrates are expensive, and there is a global shortage. The synthetic vasopressin analogue desmopressin acetate (1 deamino-[8-D-arginine]-vasopressin, DDAVP) increases plasma concentrations of coagulation FVIII and von Willebrand factor (vWF) two fold to six fold through endogenous release. The drug is an attractive therapeutic alternative because it carries no risk of transmission of infectious diseases. Desmopressin is today a widely used hemostatic agent not only in patients with mild hemophilia A or von Willebrand disease (vWD) but also in those with congenital or acquired platelet dysfunction. There is a long clinical experience with the drug because it has been used for prevention of bleedings in connection with invasive procedures and for treatment of bleedings since the mid-1970s. Not all hemophilia A patients can be treated. The clinical usefulness depends on the postdesmopressin plasma concentration of FVIII, which in turn depends on the patient's basal FVIII level. Therefore, a test dose is recommended in candidate patients. In general, only the mildest hemophilia A patients respond sufficiently. Optimal hemostatic effect is achieved with a dosage of 0.3 microg/kg given intravenously. An intranasal desmopressin spray is suitable for the home treatment. PMID- 12640573 TI - Gene transfer as an approach to treating hemophilia. AB - Gene therapy is a novel area of therapeutics in which the active agent is a nucleic acid sequence rather than a protein or small molecule. Successful clinical applications of gene transfer have been limited to date because of shortcomings in the available gene delivery vehicles. The goal of gene transfer for hemophilia is to achieve sustained expression of factor (F) VIII or FIX at levels high enough to improve the symptoms of the disease. Hemophilia has proved to be an attractive model for those interested in gene transfer, and multiple gene transfer strategies are currently being investigated. So far, five different trials, three for hemophilia A and two for hemophilia B, have enrolled approximately 40 patients with severe hemophilia. This article summarizes the gene transfer strategies being investigated, the available preclinical data, and the early clinical results. In the past year, several groups have demonstrated sustained expression of clotting factors at levels of 5 to 10% of normal in large animal models of hemophilia. The goal of the ongoing clinical studies is to determine whether these results can safely be extended to humans. PMID- 12640575 TI - Endoscopic surgery of the lumbar epidural space (epiduroscopy): results of therapeutic intervention in 93 patients. AB - Determination and therapy of the underlying pathology in chronic pain syndrome in the lumbar spine is frequently difficult. Minimally invasive and microsurgical techniques may offer advantages. Epiduroscopy is available for visualization of the lumbar epidural space. 93 patients with chronic back-leg pain syndrome were epiduroscopically operated. When findings were appropriate, mechanical instruments and the holmium:YAG laser were applied therapeutically. 45.9 % of these patients presented with positive results in postoperative examination. Pathomorphological processes corresponding to the multifactorial pain processes, which escape detection in modern imaging procedures, can be diagnosed in the epidural space using epiduroscopy Therapeutic intervention is basically possible. However, use is limited due to technical difficulties. Navigation of the endoscope is especially limited in access via the hiatus sacralis. PMID- 12640576 TI - Image-guided stereotaxy in the interventional MRI. AB - BACKGROUND: Stereotactic procedures employing frame-based systems and utilizing pre-operative MR or CT have several shortcomings such as long procedure time, patient discomfort and transport, poor fail-safe capabilities and targeting inaccuracies due to brain shift. Conducting all procedural steps in an interventional MRI has the potential of alleviating some of these deficiencies. METHODS: A stereotactic system consisting of a skull-mounted mechanical positioning device and customized navigation software has been developed. The accuracy of this system was tested within an interventional MRI employing a skull phantom. RESULTS: The mean distance between the targets hit and the planned target coordinates was 0.70 mm +/- 0.3 mm with a maximum distance of 1.3 mm. INTERPRETATION: The results indicate that the proposed stereotactic system can be used for stereotactic procedures in the interventional MRI. PMID- 12640577 TI - Endoscopic release in carpal tunnel syndrome: analysis of clinical results in 200 cases. AB - Carpal tunnel syndrome is the most common peripheral neuropathy. Conventional carpal tunnel surgery has been performed as a primary procedure for the decompression of the median nerve at the wrist in patients who have idiopathic carpal tunnel syndrome. While the results have been excellent, this surgical procedure has been reported to be related to high postoperative morbidity and extended length of recovery time. Over the past decade, endoscopic release of the transverse carpal ligament has been developed as a new, alternative method to the open procedures. Endoscopic carpal tunnel release has been reported to ensure less postoperative morbidity, more rapid recovery of strength, with earlier return to work, reduced disability time and a better cosmetic result. The authors present a surgical series of 200 hands in 164 patients (36 bilaterals) with idiopathic carpal tunnel syndrome, who underwent a single-portal endoscopic carpal tunnel release (Agee technique), with regards to the clinical outcome and complications occurred after 4-months follow-up. PMID- 12640578 TI - The role of third ventriculostomy in the management of obstructive hydrocephalus. AB - OBJECTIVE: Endoscopic third ventriculostomy (ETV) is an effective treatment for occlusive hydrocephalus caused by an obstruction of the CSF flow in the aqueduct or the posterior fossa. We evaluated the factors age, pathology and surgical technique on the results of the ETV. METHODS: Between November 1992 and October 2000 171 ETV have been performed in 159 patients. The follow-up was evaluated in 150 patients. The age ranged from 10 days to 77 years (mean age 35 years). The hydrocephalus was caused by benign aqueductal stenosis in 77 patients, space occupying lesions in 59, by intraventricular hemorrhages in 11, and by other causes in 3 patients. The trajectory was planned in 31 patients by frame-based and in 4 patients by frameless stereotaxy. RESULTS: The overall success rate of a single ETV was 71.3 % and including successful re-ETV 76.7 %. Best results were observed in adults and children older than 1 year. Infants demonstrated significantly worser outcomes. Patients with benign aqueductal stenosis and tumor compressing the aqueduct had the greatest profit from the ETV. The stereotactic guidance had no influence on the outcome and the number of severe complications. Complications were one arterial bleeding, three venous bleedings, and one ICB, all without permanent deficit, except one permanent hemiparesis. No mortality was observed. PMID- 12640579 TI - End-to-side anastomoses in small arteries with partial lateral clipping. An experimental study. AB - The problem of total therapeutic occlusion of the cranial arteries remains a relatively important area of concern for neurosurgery, neurology, neuroradiology and neuroanesthesia as well as for the patients. Cerebral revascularization procedures require total occlusion of the cranial arteries for a while. End-to side anastomoses with partial lateral clipping were done in 20 cadaver arteries and then, in one rat common carotid artery. The results of the study were compared with the current cerebrovascular bypass procedures on a theoretical basis and possible clinical implications of the method were suggested. All the anastomoses were found to be open. As conclusions; 1) End-to-side anastomosis with partial lateral clipping and along with this technique the maintenance of blood flow in the target vessel during the anastomosis procedure are experimentally possible. 2) Although the available aneurysm clips may be used in the bigger vessels (> 2.5 mm), currently, the exclusive clips for the technique are not available commercially. 3) Further clinical implications of the technique may be investigated. PMID- 12640580 TI - Anatomic variations of the frontal sinus. AB - INTRODUCTION: Minimally invasive approaches are increasing in popularity. We have undertaken an anatomic and radiological study of the frontal sinus that is frequently involved in the supercilliary minicraniotomy used to access the parasellar region. OBJECTIVE: We wanted to evaluate the prevalence and morphological characteristics of the frontal sinus in a certain population sample. METHODS: 98 randomly assigned individuals were subjected to CT examinations. The antero-posterior diameter, sagittal diameter, transverse diameter and total volume of the frontal sinus were calculated. RESULTS: The frontal sinus tended to be larger in males than in females with the exception of the transverse diameter (p < 0.10). CONCLUSION: Knowledge of the anatomic variations of the frontal sinus is important in surgical approaches through the superciliary arc in order to avoid complications such as infections and CSF fistula. PMID- 12640581 TI - Peri-operative levels of s-100 protein in serum: marker for surgical manipulation and postoperative complications. AB - Although minimally invasive neurosurgical techniques are highly sophisticated nowadays, almost any operative procedure causes an inevitable surgical trauma to the brain. As a consequence unfavorable functional outcomes are not rare. Intraoperative biochemical monitoring can be helpful first to detect but also to prevent brain damage. We investigated if serum S-100 protein (S-100) levels are a reliable marker for the extent of acute cerebral damage caused by surgical trauma or postoperative complication. S-100 is present in the cytosol of glial cells. This protein leaks into the extracellular space after cell damage and can be detected both in the cerebrospinal fluid (CSF) and serum. To determine S-100 protein levels, serum samples from 20 patients with various intracranial tumors were collected before surgery, and at one day, as well as at seven days after surgery. It was hypothesised that the size of the tumor-brain contact surface (TBCS) was closely related to the dimension of the surgical trauma. TBCS was measured from radiological imaging. The pre- and postoperative (day 1 and day 7) clinical condition of each patient was assessed. The S-100 levels were correlated with the TBCS and the clinical condition. Levels of S-100 on day 1 and day 7 were significantly higher as compared with levels on day 0 ( p = 0.02, respectively p = 0.01). There was a significant relationship between rise of S-100 level and worsening of clinical condition between day 0 and day 1 ( p = 0.001). Also a significant positive relationship between TBCS and the level of S-100 could be found on day 1 and on day 7 ( R = 0.71, p = 0.0009, respectively R = 0.73, p = 0.004). Furthermore, a significant relationship between the rise of S-100 level between day 0 and day 1, as well as between day 0 and day 7, and TBCS could be documented ( R = 0.61, p = 0.01, respectively R = 0.64, p = 0.005). In conclusion, serum S-100 levels are a reliable marker for acute or recent CNS damage caused by neurosurgical manipulation or as a result of secondary postoperative complications. Therefore, intraoperative monitoring of serum S-100 levels seems very promising. In such a setting the negative effects of surgical manipulation can be measured instantaneously, which should bring the neurosurgeon to change his strategy. As a consequence the surgical trauma can be minimized and functional outcome can be optimized. PMID- 12640582 TI - Improving planning procedure in brain biopsy: coupling frame-based stereotaxy with navigational device STP 4.0. AB - 37 consecutive patients with space-occupying intracerebral lesions were operated via frame-based stereotaxy. After CT-localizing of suspect lesions and computer supported definition of entry and target coordinates a serial stereotactic biopsy was performed. Biopsy specimens allowed a satisfactory neuropathological examination and diagnostic result in 36 cases (97 %). Only three patients (8.1 %) showed an intraoperative bleeding, which was not associated with any postoperative CT-detectable hematoma, neurosurgical intervention nor with any neurological deficits. In summary we described the method of a computerized planning technique for stereotactic biopsy with the use of a special stereotactic planning program. High percentage of satisfactory neuropathological diagnoses and comfortable and accurate definition of target and entry coordinates justify computer support as a routine method. PMID- 12640583 TI - Capillary haemangiomas of the spinal cord: report of two cases. AB - The clinicopathological features of two cases of capillary haemangioma of the spinal cord are described. The presenting symptoms were similar to those of common intradural, intramedullary tumours or tumour-like mass lesions. Radiological features of these two lesions resembled other vascular lesions of the spinal cord. The patients underwent surgery, and recoveries were good. Histologically, the lesions resembled capillary haemangioma of superficial tissues that are composed of lobules of small capillaries with feeding vessels. A fibrous capsule enveloped the lesions. Capillary haemangiomas of the central nervous system are rare. Awareness of their existence may help the surgeon to avoid misdiagnosis and overtreatment of these benign tumour-like lesions. PMID- 12640584 TI - Trigeminal neuralgia presenting as Chiari I malformation. AB - Chiari I malformation usually presents with headache, numbness, weakness and gait abnormalities. We present a 38-year-old female with trigeminal neuralgia who was found to have a Chiari I malformation. She had a foramen magnum decompression with complete resolution of her symptoms. PMID- 12640585 TI - Schwannoma of the lateral ventricle: eight-year follow-up and literature review. AB - Intracerebral and intramedullary schwannomas are uncommon; but, in general, spinal intramedullary schwannomas are more frequent than intracerebral schwanomas. We present a case of right lateral ventricle schwannoma in a 21-year old man and review the associated literature. The 21-year-old right-handed man presented with loss of the left-eye vision approximately 8 months before referral to an ophthalmologist. The patient was immediately subjected to computed tomography (CT) scan, which showed an enhanced lesion with cystic component in the right occipital horn of the lateral ventricle. And consecutively, he was admitted to our department. The tumor was evacuated via craniotomy with marked improvement in his clinical state. The postoperative course was uneventful and postoperative CT control showed no residue. On MRI control no recurrence was noted after a follow-up period of 8 years. Intracerebral schwannoma is a rare, benign neoplasm. It is usually located superficially or adjacent to a ventricle. Characteristic imaging features include cyst formation, calcification, and evidence of peritumoral edema or gliosis. The recognition of this benign and potentially curable neoplasm and its differentiation from other neoplasms, some of which have less favourable outcomes, is of obvious importance. PMID- 12640586 TI - Endoscope-assisted microsurgery of the corpus callosum. AB - The authors describe the use of a rigid endoscope lens to enhance microsurgical visualization of the corpus callosum. Using cadaver preparations, endoscopic dissection was carried out through a narrow corridor without significant retraction of the brain. The endoscope improves the operative exposure in corpus callosotomy and other transcallosal procedure. PMID- 12640587 TI - Stereotactically-guided fourth ventriculo-peritoneal shunting for the isolated fourth ventricle. AB - The isolated fourth ventricle (IFV) develops in which obstruction to the out flow of cerebrospinal fluid from the choroid plexus of the fourth ventricle occurs rostrally and caudally. IFV has been a rare occurrence and is difficult to treat. We had an occasion to admit a 28-year-old female to our hospital due to hydrocephalus: she also had a history of meningitis a year ago. The patient was initially managed by a lateral ventriculo-peritoneal shunting procedure. Six months after the procedure the patient began to suffer from vomiting, nausea, and diplopia. CT and MRI scans demonstrated an isolated fourth ventricle enlargement; and thus, a fourth ventriculo-peritoneal shunting procedure was performed under stereotactic conditions. The authors present a case of an isolated fourth ventricle after lateral ventriculo-peritoneal shunting for hydrocephalus, which was treated with a stereotactically guided fourth ventriculo-peritoneal shunting procedure. The technique of this procedure is described below. PMID- 12640588 TI - A new device for electrical stimulation mapping of the brainstem and spinal cord. AB - With the goal to minimize the risk of sequelae in brainstem and spinal cord surgery, methods of intraoperative mapping using direct electrical stimulations were developed in the last decade. However, the current was delivered with a monopolar stimulator with risk of a spreading, or using a bipolar probe or silicone strip with 5 mm of precision. To improve the accuracy and easiness of the mapping, the authors describe in this work a new device with the following characteristics: bipolar probe without current diffusion; reduction of the space tips to 2 mm; building of a bent handle, improving the field of view and ergonomics under the microscope. This device has been used successfully in 6 consecutive patients, 4 harboring a brainstem lesion and 2 with a spinal cord tumor. This new device allows the surgeon to perform repetitive intraoperative brainstem and spinal cord electrical mapping under the microscope in a very simple, quick and accurate way all along the resection. Improvement of such a mapping first minimizes the risk of sequelae, second optimizes the quality of resection, and third may also give a prognostic value concerning the changes of postsurgical recovery in case of a preoperative deficit. PMID- 12640589 TI - [Effects of quetiapine at low doses on psychosis motor disability and stress of the caregiver in patients with Parkinson's disease]. AB - INTRODUCTION: Quetiapine is a novel neuroleptic drug with pharmacological properties close to clozapine, the most effective drug in the management of psychotic symptoms in patients with Parkinson s disease (PD). Unlike clozapine, however, quetiapine does not induce agranulocytosis and therefore no haematologic controls are required. OBJECTIVES: To assess tolerability and effectiveness of low dose quetiapine on psychotic symptoms, sleep disturbances and stress of the caregiver in PD patients with dopaminomimetic psychosis. PATIENTS AND METHODS: We carried out a 16 week, prospective, open label study on the effects of quetiapine in 7 consecutive PD patients with psychosis. A ceiling dose was arbitrarily established at 25 mg/d. Antiparkinsonian medication remained unchanged throughout the study. Motor symptoms were assessed with UPDRS motor subscale portion, Schwab England scale, and Hoehn Yahr diseases staging. The effects on hallucinations and paranoia, sleep disorder, and stress in the caregiver were scored separately, aside from a global score on the Neuropsychiatric Inventory (NPI). Global impression of both, investigators and caregiver was obtained at study conclusion. A patient died from unrelated causes. RESULTS: Under a mean quetiapine dose of 24.9 mg/d (12.5 37.2) motor impairment during on periods remained unchanged, both according to patients and caregiver whereas mean UPDRS motor score remained unchanged (35.5 4.5 versus 32.8 5.2). Levodopa induced dyskinesias disappeared in the single patient with this complication. Improvement occurred in mean NPI global score, as well as subscores for hallucinations and paranoia, and caregiver stress. In over half the patients, effectiveness was scored as very good or good both by examiners and caregivers. A confusional episode occurred in one patient under 25 mg/d but readily resolved while maintaining the benefit following dose reduction to 12.5 mg. CONCLUSION: Quetiapine at low doses appears a useful alternative for psychotic symptoms, sleep disorders and stress of the caregiver in patients with PD PMID- 12640590 TI - [Aetiological diagnosis of cerebral infarction in a county hospital]. AB - INTRODUCTION: Cerebral infarction (CI) can be classified aetiologically in several different ways using explicit diagnostic criteria. However, the extent to which these diagnostic criteria are actually implemented in clinical practice is unknown. Aims. The aim of this study was to analyse the management and use of diagnostic tests in the aetiological diagnosis of CI in two county hospitals and to compare this with the most common recommendations. We also sought to analyse the clinical and demographic variables that may help to explain why these guidelines are not followed. PATIENTS AND METHODS: We reviewed the discharge abstracts of 307 cases of CI attended in two county hospitals between 1999 and 2000 and we analysed the clinical data, diagnostic tests and the final diagnosis. The diagnoses were reorganised using the TOAST, Laussane, NINDS and SEN 98 classifications and we analysed the frequency with which the diagnostic tests were employed in each aetiological subtype. RESULTS: Average age: 71.3 years; 59.3% were males. CAT scans were performed in 97.1% of cases, neurosonology was used in 40.1% and echocardiography was performed in 8.5%. The aetiological diagnosis was: atherothrombotic 22.4%, cardioembolic 10.7%, lacunar 26%, unusual causes 0.3% and unknown causes 1.6%. In 37.4% of cases the diagnosis was given as unspecified CI. On reclassifying the diagnoses according to SEN 98 criteria, we obtained the following: atherothrombotic 19.5%, cardioembolic 2.8%, lacunar 13.7% and of unknown origin 63.5%. 0.6% of the cases were unclassifiable. Factors that exerted an influence on the fact that diagnostic tests were less frequently carried out included age, level of awareness and mortality. The most frequent cause of incomplete studies was the absence of carotid Doppler. CONCLUSIONS: The guidelines for aetiological diagnosis of CI are not often followed. Systematic performance of a neurosonological study would improve aetiological diagnosis of CI. PMID- 12640591 TI - [Migraine and MIDAS (MIDASELA) in Colombian hospital workers]. AB - AIMS: The aim of this study was to determine the prevalence of migraine and its implications in the occupational and outside employment/daily activities of the workers at a regional hospital in the Cundiboyacense Plateau in Colombia. PATIENTS AND METHODS: The available members of the house staff at this institution were interviewed by applying the neuroepidemiological protocol drawn up by the World Health Organization (WHO); general doctors performed the initial screening and the determination of neurological disease was carried out by a clinical neurologist, both in patients who were positive and negative for neurological disease. Quality of life was evaluated by means of the MIDAS (Migraine Disability Assessment) survey, MIDASELA (in Spanish for Latin America); the analysis was performed using the EPI 6.04 software application. RESULTS: A total of 238 people were studied: 188 females and 50 males; the prevalence of migraine was 22.5% (15.5% by the WHO protocol and 7% false negatives), with predominance in females (OR: 5.49; p< 0.005). In the MIDASELA questionnaire, 47.2% of the patients had a 50% alteration in their productivity, with regard to their occupational and outside employment/daily activities. A minimum number of patients (3.8%) were unable to work because of migraine, as compared to the figures for outside employment/daily activities (29.7%) or those involving leisure/family (66.4%). The average work, outside employment/daily and family time lost per patient because of migraine was 0.3, 2 and 5.2 days, respectively. CONCLUSION: A sub register in the prevalence of migraine could be reported in places where the WHO protocol is used. This pathology will have to be taken well into account in Colombian hospital workers due to its individual, occupational, family, social and economic impact. It is necessary to introduce health policies and programmes aimed at evaluating, controlling and treating this type of pathology in an appropriate manner, in order to improve the quality of life of those whose job it is to look after and improve that of the other Colombians. PMID- 12640592 TI - [A statistical analysis of 1300 patients with dizziness-vertigo. Its most frequent causes]. AB - PATIENTS AND METHODS: We analysed the records of the individuals who were attended because of dizziness or vertigo in the vestibular sector, with the aim of describing the epidemiological clinical profile of a group of patients with such symptoms. 1300 patients were systematically evaluated according to our neuro otological examination protocol. Diagnoses were ordered, according to the international classification reported by Drachman and later modified by Bahlo, in four categories: 1. Vertigo, 2. Instability, 3. Pre syncope and 4. Miscellaneous. Each of these classes was organised according to the topography of the lesion and these were in turn grouped by aetiologies (viral, vascular, tumoural, demyelinating, post traumatic, idiopathic, autoimmune, etc.). The data were stored and analysed in a computer database, Epi info 6.02 (OMS 1994), which was especially adapted by the researchers for the purpose. RESULTS: 63.1% were women. The average age was 55.5 years old (SD: 17.5, interval: 4 93). Vertigo was diagnosed in 68.9%, instability was found in 12.4%, 1.8% presented syncope and miscellaneous disorders occurred in 16.9% (of these, 64.1% had disorders of the central integrator and 16.4% were of a psychogenic origin). Of the 1300 patients, 896 presented vertigo; the positional type was seen in 54%, sustained in 6.5%, recurrent in 27.7% and 11.8% were found to have the otolithic type. CONCLUSIONS: The relevance of the epidemiological work based on clinical evaluation and the thorough neuro otological examination in our medium must be highlighted. These findings were similar to those reported in the international literature in more delimited series. PMID- 12640593 TI - [Tolerance and adverse events of treatment with acetylcholinesterase inhibitors in a clinical sample of patients with very slight and mild Alzheimer s disease over a six-month period]. AB - INTRODUCTION: At present acetylcholinesterase inhibitors (AChEI) are used in the treatment of the cognitive deterioration associated with Alzheimer s disease (AD). The side effects of these drugs are linked with the increase in acetylcholine, which limits their effectiveness, and must be adjusted to the patient close to the maximum tolerated dose. PATIENTS AND METHODS: We conducted a comparative retrospective study of the tolerance and the adverse events (AE) of two AChEI in a group of patients with very slight and mild probable AD over a 6 month period. RESULTS: The sample was made up of 175 patients, of which 134 began therapy with 5 10 mg/day of donepezil and 41 with 6 12 mg/day of rivastigmine. 20% of the patients presented AE and 8% abandoned the treatment. Gastrointestinal disorders (GID) were the main AE observed (57.1%). Only 6% of the patients treated with donepezil abandoned the therapy because of the AE as opposed to 14.6% of the patients treated with rivastigmine. Patients treated with rivastigmine displayed a higher incidence of GID and the relative risk of presenting GID was 4.4 times higher than in the patients treated with donepezil. CONCLUSIONS: The GID associated to therapy with AChEI are the main reason for abandoning treatment and occur more frequently in patients treated with rivastigmine. PMID- 12640594 TI - [Cognitive evoked potentials in autistic children]. AB - AIMS: The aim of this study was to examine the latency, amplitude and distribution of N400 potential in order to evaluate the semantic processing capacity in autistic children and in children suffering from Asperger s syndrome (AS), and to compare them with a control group. PATIENTS AND METHODS: 24 autistic children, six boys with AS and 25 controls, aged between 6 and 14 years old. The cases were examined using the DSM IV diagnostic criteria. Auditory stimulation was performed with pairs of congruent and incongruent words: two lists of 20 pairs of semantically related words (congruent) and 20 pairs of words with no semantic relationship whatsoever (incongruent). RESULTS: The most striking parameter is the increase in latency in N400 for the group of autistic children, which did not occur in the group of children with AS. Maximum N400 negativity for the children with autism was found in the left frontocentral region. No significant differences were observed for the amplitude of N400 between the three groups that were studied. CONCLUSION: Neurophysiologically, the autistic children and those affected by AS perhaps use different neuronal networks in semantic processing. The N400 wave can be a valid test for monitoring verbal processing in these children. PMID- 12640595 TI - [Reflex myoclonic epilepsy in infancy: a new reflex epilepsy syndrome or a variant of benign myoclonic epilepsy in infancy]. AB - CASE REPORTS: We report a clinical and EEG study of 8 children with reflex myoclonic epilepsy of infancy to further confirm the existence of this syndrome first described by Ricci et al in 1995. RESULTS: Between February 1990 to July 2002, we identified 64 epileptic patients with myoclonic seizures with an onset in the first six years of life. Eight (12.5%) of these patients had myoclonic seizure stimuli sensible. The seizures were characterized by generalized, myoclonic jerks triggered by tactile stimuli in six patients and acoustic stimuli in two, in one of them myoclonic jerks were triggered by both types of stimuli. The seizures appeared between 5 and 20 months of age. Two of the 8 patients had spontaneous myoclonic attacks during sleep. Interictal EEG was normal during wakefulness and occasional discharges were evident during sleep. In contrast, the ictal EEG during both wakefulness and sleep showed generalized spike wave and polyspike slow wave paroxysms. Neurologic examination, neuroimaging and neurometabolic studies were normal. Myoclonic jerks disappeared in 6 patients after valproic acid administration and in two after clobazan administration. Antiepileptic treatment was discontinued in 6 patients and no seizure recurrence was observed during a median follow up of 6 years. CONCLUSION: Our patients presented electro clinical criteria compatible with the syndrome of reflex myoclonic epilepsy of infancy. This syndrome could be considered to be a new reflex epileptic syndrome or a variant of benign myoclonic epilepsy in infancy. PMID- 12640596 TI - [Can treatment associated with ticlopidine and nifedipine increase serum levels of phenobarbital?]. AB - INTRODUCTION: We report a case in which the association between ticlopidine, nifedipine and phenobarbital was linked with a higher than expected phenobarbital concentration in serum, which suggested a possible interaction between these drugs. CASE REPORT: A 67 year old male who received treatment with phenobarbital, digoxin, nifedipine, ticlopidine, paroxetine and clorazepate dipotassium. The first control of the level of phenobarbital in serum was performed without any symptoms or signs of toxicity or ineffectiveness. A phenobarbital concentration in serum of 21.4 mg/L was obtained, with a serum level/dosage ratio of 16.7. DISCUSSION: The serum level/dosage ratio of phenobarbital that was found in this case is almost twice as high as expected. In the absence of other factors that can explain this finding, we believe that two drugs (ticlopidine and nifedipine) may be involved in an interaction with phenobarbital. CONCLUSIONS: The high value of the serum level/dosage ratio that was found makes it advisable to monitor the concentrations of phenobarbital in serum in treatment associated with ticlopidine or nifedipine, especially when adjusting the dosage, beginning or ending treatment with these drugs. PMID- 12640597 TI - [Giant atherosclerotic aneurysm. A case report]. AB - INTRODUCTION: Intracraneal aneurysms are frequent in medical practice, most of them never produce symptoms, that is why occasionally, they are not diagnosed before death. Sacular is the most frequent aneurysm, and atherosclerotic aneurysm is rare. CASE REPORT: In this article we present the case of a 75 year old woman who has been suffering from high blood pressure and coronary artery disease since more than 20 years and in the last four months she had been complaining of dysphagia and dysartria. She was taken to the hospital because 24 hours ago mild headache started accompanied by vertigo, nausea and vomiting. On neurological exam we found sleepiness, dysartria, and right braguio crural hemiparesis, therefore we decided admitt her, but after 48 hours left braquio crural hemiparesis was added and later respiratory sepsis appeared. The patient died due to an acute respiratory failure. CONCLUSION: Necropsy revealed a huge atherosclerotic intracraneal aneurysm (31 mm) located on basilar artery. The case has been discussed and images were shown. PMID- 12640598 TI - [Functional heterogeneity of the piramidal system: corticobulbar and corticospinal tracts]. AB - INTRODUCTION AND OBJECTIVE: The pyramidal system is constituted by the cortical fibers reaching the bulbar pyramids at the level of the inferior Olive. Close to 75% of these fibers decussate terminating at bulbar level about half of them while the other half form the crossed corticospinal tract. The majority of the uncrossed fibers descend to the cord forming the direct corticospinal tract. This revision presents evidence demonstrating the functional heterogeneity of the pyramidal system. DEVELOPMENT: The corticobulbar fibers are slower conducting and branch more than the corticospinal fibers. The conduction speed of the corticospinal fibers increases as they reach more caudal segments, thus assuring that the different signals modulating the inter segmental activity reach their targets simultaneously. Those from the somatosensory cortex end in the dorsal horn while those from the primary motor cortex end primarily on interneurons of the reflex pathways to distal limb muscles although in rats, primates and humans about 10% of them terminate directly on motoneurons. CONCLUSIONS: The corticobulbar component of the pyramidal system selects the adequate motor synergies and modulates other descending systems to coordinate the activity of the distal musculature and the associated postural adjustments. The corticospinal component is functionally related to motor activities requiring accuracy, and to motoneuronal recruitment to adjust the contractile force. Both pyramidal subsystems modulate the processing and integration of the ascending somatosensory information generated by the movement itself. PMID- 12640599 TI - [The value of topiramate used with conventional schemes as an adjunctive therapy in the treatment of Lennox-Gastaut syndrome]. AB - INTRODUCTION: Lennox Gastaut syndrome (LGS), which appears in children aged between 2 and 8 years old, is characterised by a triad of epileptic seizures with different patterns, variable degrees of mental retardation, an electroencephalogram (EEG) with slow spike wave complexes at 1.5 4 Hz and bursts of rapid centrotemporal activity, with a variable response in the control of the epileptic seizures. It requires polytherapy with associations between conventional and new antiepileptic drugs, including topiramate, with variable results in the control of seizures, especially in this syndrome, which has no known response in Mexico. Aim. To determine how it responds when topiramate (TPM) is associated with another antiepileptic drug, in order to diminish the recurrence of seizures by 50% and to improve the quality of life of Mexican children. METHOD: The study was conducted in a sample of 15 children aged between 2 and 15 years old who had been diagnosed as suffering from LGS and who used more than three conventional antiepileptic drugs. After obtaining prior authorisation from the person in charge, the frequency, duration and clinical patterns of the seizures were evaluated. Since the dosages used and the serum levels, which should be within those considered to be therapeutic, were already known for each of the subjects, we administered a Quolie 10 survey before beginning with TPM. Those who presented no modifications in the frequency and duration of the seizures over a two month period were included and treatment began gradually with 2 mg/kg/day and rose to 10 mg/kg/day as the maximum dosage. Once the frequency and duration had diminished by more than 50%, the decision was made to stop more than two antiepileptic drugs, but preferably to continue with the valproic acid. The result obtained from 15 children was a remission in over 50% of the cases and an improvement in the quality of life of the children, despite the fact that the duration ranged from six months to a year. CONCLUSION: In this group of patients TPM is useful for the control or remission of seizures and, consequently, we suggest it should be administered in the Mexican population associated with valproic acid or new antiepileptic drugs. PMID- 12640600 TI - [Methodology of basic research in cognitive neuropsychology]. AB - INTRODUCTION AND METHOD: The issue regarding the relations between behaviour and the brain is no longer the specific goal of neuropsychology, but rather that of the coordinated multidisciplinary activity of all the disciplines that go to make up the neurosciences. The specific contribution of neuropsychology to this multidisciplinary activity consists in trying to determine the nature of cognitive processes so that, in the future, it will be possible to ascertain the anatomophysiological foundations for each of them. The aim of basic research in cognitive neuropsychology is precisely that. The growing interest in research in this discipline that has been observed over the past few years in Spain has not usually been accompanied by the awareness of the fact that conducting research with damaged brains requires (if it is to be valid) a methodology that is specific and different to the methodology used in research with intact brains. In this study we describe some of the main fundamental principles of basic research in cognitive neuropsychology. Through the scientific analysis of a recent piece of research work, we illustrate the consequences of not bearing these principles in mind. CONCLUSIONS: We propose a methodology that can be used to approach basic research in cognitive neuropsychology that allows these undesirable consequences to be avoided. This same methodology constitutes a guide that enables us to evaluate the conclusions of the research, regardless of the statistical processing of the data that support them. Scientific criticism contributes more to the advance of science than actual research does. PMID- 12640601 TI - [The importance of antiphospholipid antibodies in strokes]. AB - INTRODUCTION AND METHOD: The study of antiphospholipid antibodies has aroused a great deal of interest among researchers, as some of them are related to neurological diseases and in particular with cerebral ischemia. Cases of strokes in which the aetiology is unknown have been reported in young patients. Until now anticardiolipin antibodies and lupus anticoagulant have received the most attention in studies, but recently descriptions have been published of anticardiolipin antibodies with other particularities that can act as more specific immunity markers for strokes of undetermined origin in young adults. Recent research has been directed towards gaining an understanding of the mechanisms that allow these antibodies to play a direct role in the physiopathology of thrombosis and how certain risk factors smoking, high blood pressure, lipid disorders can exert an influence on the expression of phospholipids in the cerebral endothelium. Several authors have described different pathological mechanisms that help to understand the heterogeneous clinical manifestations of antiphospholipid syndrome. CONCLUSION: There is a need to study the different possible antigens of the antiphospholipid antibodies, as well as their specificities and pathological mechanisms, in greater depth in order to obtain a phospholipid immunity marker that is useful in the diagnosis of young stroke patients who are positive for these antibodies. PMID- 12640602 TI - [Brain electromagnetic tomography distinguishes primary generalised discharges from secondary bilateral synchrony]. PMID- 12640603 TI - [Multiple cavernomatosis. Differential diagnosis by gradient-echo]. PMID- 12640606 TI - The seriously ill hospitalized patient: preferred role in end-of-life decision making? AB - PURPOSE: The objective of this study was to further our understanding of the decision-making process near the end of life. Specifically, we ascertained the seriously ill patients' preferred role in the decision-making process, what factors were associated with this role, and how this stated preference related to physicians' perception of preferred role. MATERIALS AND METHODS: Prospective cohort study of hospitalized patients with end-stage congestive heart disease, chronic pulmonary disease, cirrhosis, or metastatic cancer. Eligible patients were interviewed to ascertain their personal views on end-of-life decision making, desired role, and level of symptoms experienced. RESULTS: A total of 135 patients were enrolled in this study. The majority of patients (103, 76%) had thought about end-of-life issues although only 48 (36%) had discussed them with their doctor in the hospital. With respect to preferred role in decision making, in the scenario of a competent patient, 14 (10%) preferred to leave all decisions to the doctor, 12 (9%) preferred that the doctor make the final decision after considering their opinion, 43 (32%) preferred that the doctor shared responsibility with them to make the decision, 32 (24%) patients preferred to make the final decision after considering the doctor's opinion, 21 (16%) preferred to make the treatment decision alone, and 13 (10%) did not answer. Physicians were not able to accurately predict patient's preferred role nor could the variability in patient choice be accounted for by demographic or symptom covariates. CONCLUSION: Seriously ill hospitalized patients desire to discuss end of-life issues with their physicians but their preferred role in decision making is variable and difficult to predict. PMID- 12640607 TI - End-of-life intensive care unit decisions, communication, and documentation: an evaluation of physician training. AB - PURPOSE: The majority of patients dying in intensive care units (ICUs) do so after the forgoing of life-sustaining therapies (FLST). Communication between physicians, patients, and their families regarding the decision to FLST has not been evaluated in Israel. MATERIALS AND METHODS: All patients who had FLST in a general ICU were enrolled in the study. We evaluated whether physicians communicated and documented the FLST decisions with patients or the patients' families. We also assessed the effect of the physician's geographic place of training on communication behavior. RESULTS: Over a period of 8.5 months, 385 patients were admitted to a general ICU in Israel. Fifty-seven patients died or had FLST. Twelve of these 57 were excluded from the study. Thus, 45 (79%) patients had FLST and were enrolled in the study. All patients were deemed medically incompetent to make FLST decisions. In 24 (53%) patients, FLST was discussed with the family before the decision to forgo therapy. Discussion occurred later with 6 other families, who were unavailable at the time the FLST decision was made. In 15 patients, there were no discussions with families. American-trained physicians discussed FLST with 22 of 29 families initially and 5 other families later (93%), whereas the Eastern European-trained physicians discussed FLST with only 3 of 16 (19%) families (P <.001). Documentation of FLST was present in 26 (90%) patients of American-trained physicians and 8 (50%) patients of Eastern European-trained physicians (P <.001). CONCLUSIONS: FLST is common in an Israeli ICU. Patients are not medically competent to make FLST decisions. American-trained physicians discuss and document FLST more often than Eastern European-trained physicians. PMID- 12640609 TI - Why are critical care clinicians so powerfully distressed by family demands for futile care? PMID- 12640610 TI - Seasonal bed closures in an intensive care unit: a qualitative study. AB - OBJECTIVE: To describe perceptions of the administrative procedures for seasonal bed closures and their consequences in the intensive care unit (ICU), and to critique this example of health care priority setting for legitimacy and fairness. DESIGN: A qualitative study using case study methods and interviews with key participants. We evaluated fairness and legitimacy of the bed closure process using 4 domains of the ethical framework of "accountability for reasonableness." SETTING: An university-affiliated medical/surgical ICU in Eastern Canada. PARTICIPANTS: ICU clinicians (9 bedside nurses and 5 physicians), and administrators (3 ICU managers and 2 senior hospital executives). MAIN OUTCOME MEASURES: Perceptions of ICU clinicians and administrators regarding the ICU bed closure decision-making process and its consequences. RESULTS: Emerging themes concerned: (1) bed closure rationale (including arbitrary decision making, bed closure masquerading as a code for a nursing shortage, and suboptimal evidence base for implementing closures); (2) bed closure process (viewed as unclear with insufficient prior publicity and inadequate subsequent review); and (3) adverse consequences (including safety issues, negative professional working relationships, and poor morale). Although an appeals mechanism existed, nurses were not available to staff reopened beds so this condition is only partially met. The relevance, publicity, and enforcement conditions for accountability of reasonableness were not satisfied, offering opportunities for improvement. CONCLUSION: Clinicians and administrators are readily able to identify shortcomings in the seasonal bed closure process in the ICU. These shortcomings should be targeted for improvement so that intensive care health services delivery is legitimate and fair. PMID- 12640608 TI - Intensive care nurses' and physicians' experiences with demands for treatment: some implications for clinical practice. AB - OBJECTIVE: This study was conducted to develop an empiric description of intensive care unit (ICU) physicians' and nurses' (participants) experiences providing life-sustaining treatments at the insistence of family members, treatments that they believed should have been withheld or withdrawn. From this description, steps to minimize or prevent their sources of distress in such situations are suggested. DESIGN: Semistructured, open-ended interviews. Participants were asked to describe cases in which treatment had been provided primarily in response to demands from family members. PARTICIPANTS: Six physicians and 6 nurses from 6 university-affiliated ICUs in Canada. All were members of a task force developing a multicenter policy to address demands for treatment, and physician members were heads of their ICUs. OUTCOME MEASURES: Systematic analysis of interview transcripts and synthesis of findings. RESULTS: Participants recalled 28 cases in which treatment had been provided at the insistence of family members. Many cases described were very distressing for both medical staff and family members. Consistently problematic areas included: (1) suffering of dying patients, (2) the marked distress of family members, and (3) a breakdown in the relationship between care providers and families. CONCLUSIONS: Conflict with family members about decisions to limit life-sustaining treatment can be very stressful for health care providers. Three important areas that give rise to distress were identified in this study. These key sources of distress should be looked for. They could be addressed by: (1) identifying to family members the importance of minimizing suffering and ongoing bodily injury of patients at risk for dying, (2) by doing so addressing directly the distress of family members by the provision of emotional support, and when appropriate directed toward helping them accept that the patient is dying, and (3) pursuing efforts to maintain or create a good relationship with family members despite disagreement about the appropriateness of continuing life-sustaining treatment. PMID- 12640612 TI - Introduction: theme articles on technology assessment in critical care. PMID- 12640611 TI - Errors in measuring blood gases in the intensive care unit: effect of delay in estimation. AB - Arterial blood gas measurement is subject to a number of potential sources of error. We investigated some of these in the intensive care unit (ICU). We audited samples for adequate volume and the presence of air and found that all samples were of adequate volume, but 40% contained bubbles or froth. We compared pulse oximeter estimations of oxygen saturation (SpO(2)) with laboratory estimates (SO(2)) from arterial blood samples, and found that there was less than a 5% chance of a difference of 5% or more. We audited the delay between sampling and processing and looked for errors arising as a result. We found that 4% of samples waited longer than 30 minutes to be analyzed in the laboratory, but that there was no correlation between delay and error in partial pressure of oxygen (PO(2)), carbon dioxide (PCO(2)), or SO(2). We performed a bench study to document the changes in PO(2) and PCO(2) over time with samples stored at room temperature and on ice. We found that samples in 1.5-mL PICO 70 syringes (Radiometer Medical A/S, Bronshoj, Denmark) were stable for PO(2) and SO(2) for up to 30 minutes either at room temperature or kept in iced water, and that changes after 60 minutes were small and unlikely to be clinically significant. PCO(2) showed a statistically significant increase after 20 minutes at room temperature, but the changes were not clinically significant. PMID- 12640613 TI - Medical informatics in the intensive care unit: overview of technology assessment. AB - Effective patient care in the intensive care unit (ICU) depends on the ability of clinicians to process large amounts of clinical and laboratory data. Recently, medical informatics applications have been developed to store and display patient information and assist clinical decision making. Despite the proliferation of these systems and their potential to improve patient care, there are no comprehensive health technology assessments incorporating considerations of safety, functionality, technical performance, clinical effectiveness, economics, and organizational implications. The objectives and methods of informatics evaluations depend on the type of application and the stage of development. Qualitative and quantitative nonrandomized evaluations of comprehensive information management systems like electronic medical records and picture archiving and communications systems should concentrate on technical and functional issues. Specific applications like clinical decision support systems and computerized patient care systems are designed to improve patient outcomes and clinical performance; randomized controlled trials (RCTs) to assess clinical effectiveness are important in their assessment. Although studies of these applications in the ICU setting are increasing, there are currently very few published randomized trials. PMID- 12640614 TI - Acute renal failure in the ICU: assessing the utility of continuous renal replacement. AB - Acute renal failure (ARF) in the ICU patient still remains a common problem and is associated with increased morbidity, mortality, and cost. Potential advantages of continuous renal replacement (CRRT), compared with intermittent hemodialysis (IHD) include enhanced hemodynamic stability, increased solute removal, and greater ultrafiltration. Although it was hoped that CRRT would lead to improvement in patient outcomes, there are few prospective, randomized clinical studies comparing this modality with conventional hemodialysis in the treatment of patients with ARF. The difficulties associated with designing such prospective studies are the complex status of the medical patients and the ethical dilemma of randomizing patients to a certain dialysis modality. At this time, there is no evidence to support the assertion that CRRT improves clinical outcomes compared with IHD. PMID- 12640615 TI - Procalcitonin as a diagnostic test for sepsis: health technology assessment in the ICU. AB - Elevation in the serum concentration of procalcitonin (PCT) is associated with systemic infection. This association has led to the proposed use of PCT as a novel biomarker of bacterial sepsis. The advantages and limitations of the American College of Chest Physicians/Society of Critical Care Medicine (ACCP/SCCM) definitions of sepsis are an important consideration that affects the impact of any diagnostic test for sepsis and these issues are discussed. Our main objective is to perform a systematic health technology assessment of PCT as a diagnostic test for sepsis. In an adult intensive care unit (ICU) population, we identify a specific and important question-can PCT accurately distinguish sepsis in patients with systemic inflammatory response syndrome (SIRS) who have a suspected infection? Likelihood ratios are calculated from published data to attempt to find the best answer. The published evidence does not support a general claim that PCT is a useful decision support tool for diagnosing sepsis in patients who have SIRS. Procalcitonin has a slightly better ability to exclude the diagnosis of sepsis. The role for using PCT testing in the ICU will continue to evolve along with our understanding and definition of sepsis. PMID- 12640616 TI - Health technology assessment in the ICU: noninvasive positive pressure ventilation for acute respiratory failure. AB - Critical care practitioners have a number of health-related technologies at our disposal to provide the best possible care for our critically ill patients. Although certain technologies may improve outcomes in the intensive care unit (ICU), many technologies are disseminated without rigorous evaluation. Health technology assessment (HTA) in critical care is a complex and dynamic process, which is a powerful tool to assess a health technology for its initial use or continued application in the ICU. This article applies an HTA framework to the use of noninvasive positive pressure ventilation (NPPV) for patients with acute respiratory failure (ARF). The strongest evidence to date supports the use of NPPV in patients with ARF caused by exacerbations of chronic obstructive pulmonary disease (COPD); the benefit for patients with acute nonhypercarbic, hypoxemic respiratory failure is less clear. The success of NPPV technology depends on operator education and experience. The cost effectiveness of NPPV has been evaluated in patients with ARF caused by COPD, and cost reduction is attributed to the prevention of ventilator-associated pneumonia by avoiding endotracheal intubation. An HTA framework can help health care practitioners make important decisions regarding the acquisition of new technologies and the evaluation of current technologies. Careful evaluation of health technologies in the ICU should be an ongoing priority. PMID- 12640617 TI - Pneumomediastinum causing pneumoperitoneum during mechanical ventilation. AB - The causes of nonsurgical pneumoperitoneum include numerous diseases. We report here a case of nonsurgical pneumoperitoneum caused by benign pneumomediastinum during mechanical ventilation, and we discuss the anatomic pathways and the pathophysiologic mechanisms responsible for the development of this association. PMID- 12640618 TI - Fire exit is a potential four transmembrane protein expressed in developing Drosophila glia. AB - Glia from many diverse organisms play a number of important roles during the development of the nervous system. Therefore, knowing the molecules that control glial cell function will further our understanding of the mechanisms that control nervous system development. We have isolated a novel gene in Drosophila melanogaster that is expressed in a subset of the peripheral glia. We call this gene "Fire exit" (Fie), as the glia that express this gene do so during a time when they mark the entry and exit point of axons at the CNS/PNS boundary. This subset of peripheral glia act as intermediate targets during pathfinding and migration of the sensory axons in particular. Fire exit has been cloned and found to encode a novel transmembrane protein. Fire exit belongs to a group of proteins identified in the Drosophila melanogaster and Anopheles gambiae databases which contain four predicted transmembrane domains and a shared intracellular motif. Mutations that remove the fire exit protein have no obvious disruption to glial function. On the other hand, glia expressing the Fire exit gene bridge the transition zone between CNS and PNS and play a role in sensory axon guidance. Therefore, it appears that, while the glia that express this protein mediate axon guidance, Fire exit itself plays a nonessential part in this function. A role for Fire exit in glial development may be suggested by its evolutionary relationship to a family of lysosome-associated proteins called LAPTMs and suggests that Fire exit may function in intracellular transport during glial development. PMID- 12640620 TI - Versatile inducible activation system of Akt/PKB signaling pathway in mice. AB - We report a transgenic mouse line in which Akt/protein kinase B (PKB) pathway can be activated in an inducible manner in defined cell types. In this transgenic mouse line, Cre expression allows the expression of a tamoxifen-activatable form of Akt/PKB in a defined cell type. Subsequent injection of tamoxifen triggers the transient activation of Akt/PKB in mice. Thus, this transgenic line allows the transient activation of Akt/PKB pathway in a predefined cell type. We expect that this transgenic system will provide a unique tool to study the roles of Akt/PKB pathway in mice. PMID- 12640619 TI - Evidence for novel fate of Flk1+ progenitor: contribution to muscle lineage. AB - Flk1 is one of the specific cell surface receptors for vascular endothelial growth factor and one of the most specific markers highlighting the earliest stage of hematopoietic and vascular lineages. However, recent new evidence suggests that these Flk1(+) mesodermal progenitor cells also contribute to muscle lineages. All evidence is based on the experiments using in vitro differentiation and in vivo transplantation systems. Although this approach revealed a differentiation potential range of Flk1(+) cells that is wider than previously expected, it fails to determine whether Flk1(+) cells contribute to muscle lineage as part of the normal developmental process. To obtain direct evidence for the fate of Flk1(+) cells in development, we used a knock-in mouse line where Cre is expressed in Flk1(+) cells. Studies with these Cre lines provide direct evidence that Flk1(+) cells are progenitors for muscles, in addition to hematopoietic and vascular endothelial cells. PMID- 12640621 TI - New serum-free in vitro culture technique for midgestation mouse embryos. AB - Current in vitro culture methods for mouse embryos are critically dependent on specially prepared rodent serum. Rodent serum requires careful preparation and stringent assessment of serum quality, while commercially available whole embryo culture serum is expensive and shows considerable lot variability. Thus, preparation and testing of suitable serum represents a considerable investment of time and resources, particularly for laboratories with only short-term embryo culture requirements. In addition, serum supplementation of culture medium may introduce unknown serum components that could interfere with interpretation of experimental results, especially where the study is geared towards analysis of a specific growth factor. Here we describe the composition of a standardized serum free culture medium comprised of commercially available stem cell media supplements. With this method, we have successfully cultured midgestation stage mouse embryos and demonstrated, using both morphological and gene expression criteria, that these embryos exhibited proper developmental progression. We believe this method to be a significant advance in whole embryo culture technology that will be of particular use to laboratories needing to utilize whole embryo culture to study midgestation organogenesis. PMID- 12640623 TI - Tissue-specific developmental requirements of Drosophila tyrosine hydroxylase isoforms. AB - Drosophila tyrosine hydroxylase (DTH) is a key enzyme in dopamine (DA) biosynthesis, which is expressed in neural and hypodermal DA-synthesizing cells. We previously reported that two DTH isoforms are produced in flies through tissue specific alternative splicing that show distinct regulatory properties. We have now selectively expressed each DTH isoform in vivo in a pale (ple, i.e., DTH deficient) mutant background. We show that the embryonic lethality of ple can be rescued by expression of the hypodermal, but not the neural, DTH isoform in all DA cells, indicating that the hypoderm- isoform is absolutely required for cuticle biosynthesis and survival in Drosophila. In addition, we report new observations on the consequences of DTH overexpression in the CNS and hypoderm. Our results provide evidence that tissue-specific alternative splicing of the DTH gene is a vital process in Drosophila development. PMID- 12640622 TI - Efficient gene delivery into murine ovarian cells by intraovarian injection of plasmid DNA and subsequent in vivo electroporation. AB - We describe the use of direct injection of circular plasmid DNA and subsequent in vivo electroporation (EP) for efficient gene delivery to the ovarian cells, including follicular cells and oocytes of mice. When Trypan blue (TB) was injected into the central portion of an ovary by a glass micropipette, rapid dispersion of TB to each preantral and antral follicle was observed. Injections of lacZ-expressing plasmid DNA and subsequent in vivo EP resulted in transfection of follicles with efficiencies ranging from 8-60%, together with cells in the thecal portion of the ovary. Of the lacZ-positive follicles, some oocytes were also positive for lacZ activity. These findings suggest that a solution introduced inside the ovary is rapidly dispersed to each follicle. With this technique, we expect great progress in genetic engineering in murine ovary. PMID- 12640624 TI - The jewel wasp Nasonia: querying the genome with haplo-diploid genetics. AB - The jewel wasp Nasonia vitripennis is considered the "Drosophila melanogaster of the Hymenoptera." This diminutive wasp offers insect geneticists a means for applying haplo-diploid genetics to the analysis of developmental processes. As in bees, haploid males develop from unfertilized eggs, while diploid females develop from fertilized eggs. Nasonia's advantageous combination of haplo-diploid genetics and ease of handling in the laboratory facilitates screening the entire genome for recessive mutations affecting a developmental process of interest. This approach is currently directed toward understanding the evolution of embryonic pattern formation by comparing Nasonia embryogenesis to that of Drosophila. Haplo-diploid genetics also facilitates developing molecular maps and mapping polygenic traits. Moreover, Nasonia embryos are also proving amenable to cell biological analysis. These capabilities are being exploited to understand a variety of behavioral, developmental, and evolutionary processes, ranging from cytoplasmic incompatibility to the evolution of wing morphology. PMID- 12640625 TI - Analysis of an expanded width of albumin fraction by cellulose acetate membrane electrophoresis in IgA nephropathy urine before treatment. AB - Cellulose acetate membrane electrophoresis with colloidal silver stain re-vealed that the width of the albumin fraction in IgA nephropathy (IgAN) urine before treatment was significantly expanded. This phenomenon was not shown in IgAN urine after treatment or in non-IgAN urine. There was a reverse correlation between the width of the albumin fraction and the albumin con-centration in IgAN urine. By immuno-fixation, Tamm-Horsfall protein (THP) was located in the same position as the albumin band in IgAN urine before treatment; however, in the urine of a healthy subject it was located in the same position as alpha(1)-globulin. By ELISA, the THP-albumin complex concentration in IgAN urine before treatment was significantly higher than in the other two diseases. The width of the albumin fraction and the sodium ion concentra-tion of the urine were significantly correlated. The THP/albumin ratio in IgAN urine before treatment was significantly higher than in the other two groups. This suggests that the characteristic expanded width of albumin found by immunofixation indicates a THP albumin complex, and that the sodium concentration of urine is involved in the formation of this complex. PMID- 12640626 TI - Cellulose acetate membrane electrophoresis in the analysis of urinary proteins in patients with tubulointerstitial nephritis. AB - Urinary proteins from 14 patients with tubulointerstitial nephritis were analyzed by cellulose acetate membrane electrophoresis. Urinary total protein concentrations were measured, and urinary 15 proteins (prealbumin, albumin, alpha(1)-microglobulin, alpha(1)-antitrypsin, alpha(2)-macroglobulin, haptoglobin, retinol binding protein, transferrin, beta(2)-microglobulin, IgA, IgG, kappa- and lambda-light chains, cystatin C, and lysozyme) were identified by the use of a rapid and highly sensitive colloidal silver staining reagent suited for use with cellulose acetate membranes, as reported previously by Matsuda et al. (J Clin Lab Anal 15:171-174, 2001; Clin Chem47:763-766, 2001) and Hiratsuka et al. (J Clin Lab Anal 10:403-406, 1996). We also analyzed urinary total protein concentration and urinary protein fractions according to the presence of acute or nonacute interstitial nephritis. In addition, the relationship between urinary protein fraction and complications of interstitial nephritis was analyzed. The goal of this work was to find a useful index for the diagnosis of tubulointerstitial nephritis. PMID- 12640627 TI - New enzymatic assay for serum urea nitrogen using urea amidolyase. AB - We established an enzymatic assay for measurement of serum urea nitrogen using urea amidolyase (EC 3.5.1.45) from yeast species. The method is based on hydrolysis of urea by the enzyme. In this assay, we eliminated endogenous ammonium ion by use of glutamate dehydrogenase (EC 1.4.1.4). Then in the presence of urea amido-lyase, ATP, bicarbonate, magnesium, and potassium ions, ammonium ion was produced proportionally to urea concentration in serum. The concentra tion of ammonium ion formed was determined by adding GLDH to produce NADP(+) in the presence of 2-oxoglutarate and NADPH. We then monitored the change of absorbance at 340 nm. The inhibitory effect of calcium ion on this assay was eliminated by adding glyco-letherdiamine-N, N, N', N'-tetraacetic acid to the reaction system. The with-in-assay coefficient of variations (CVs) of the present method were 1.80-3.76% (n = 10) at 2.8-19.0 mmol/L, respectively. The day-to-day CVs were 2.23-4.59%. Analytical recovery was 92-115%. The presence of ascorbic acid, bilirubin, hemoglobin, lipemic material, ammo-nium ion, or calcium ion did not affect this assay system. The correlation be-tween values obtained with the present method (y) and those by another enzy-matic method (x) was 0.997 (y = 1.02x - 0.10 mmol/L, Sy/x = 0.841, n = 100), with a mean difference of -0.18 +/- 0.86 mmol/L [(values by reference method - that of present method) +/- SD] using the Bland-Altman technique. J. Clin. Lab. Anal. 17:52-56, 2003. PMID- 12640628 TI - TAP1 gene AccI polymorphism is associated with atopic bronchial asthma. AB - Asthma is a hyperresponsive airway disease that may involve inflammation responses. A transporter associated with the antigen processing 1 gene (TAP1) is involved in antigen processing, and is therefore considered to play a role in the pathogenesis of bronchial asthma. The aim of this study was to test whether the polymorphisms of the TAP1 gene are a genetic marker for susceptibility to bronchial asthma. A normal control group comprised of 43 healthy people, and 116 patients with allergic asthma were examined in this study. The polymorphism was detected by polymerase chain reaction (PCR)-based restriction analysis. Associations between atopic bronchial asthma and TAP1 polymorphisms were evaluated. The results revealed no significant differences between normal individuals and asthmatics in regard to the TAP1 gene DpnII polymorphism (P=0.752). However, there was a significant difference between the control and asthma groups as regards the TAP1 gene AccI polymorphism (P=0.020). The odds ratio (OR) of GG homozygotes of the TAP1 AccI polymorphism was 229.8 compared with the AA homozygote group. The results show that the AccI polymorphism may be an indicator for atopic bronchial asthma. PMID- 12640629 TI - Serum microminerals and the indices of lipid metabolism in an apparently healthy population. AB - Serum copper and zinc concentrations were measured in 560 apparently healthy Kuwaitis (238 males and 322 females) aged 15-80 years to assess micromineral effect on the indices of lipid metabolism. Following the recommended guidelines of the European Atherosclerosis Society (EAS) and the National Cholesterol Education Program Expert Panel (NCEPEP), the incidence of dyslipidemia was assessed from enzymatic assay data of triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) concentrations. Males had significantly lower TC (P=0.029) and HDL-C (P<0.0001) levels than females, while TG were significantly (P=0.023) lower in females. The prevalence of hypercholesterolemia, hypertriglyceridemia, elevated LDL-C, and low HDL-C levels were 35, 30, 22, and 13%, respectively. Copper did not correlate with zinc (r = -0.067, P = 0.135) but was positively associated with TC (r=0.196, P<0.0001), LDL-C (r=0.134, P = 0.003), TG (r = 0.092, P=0.039), and age (r=0.281, P<0.0001). It is concluded that unlike in animal studies, copper excess in humans is associated with hyperlipidemia and therefore will predispose to atherosclerosis. PMID- 12640630 TI - Reticulocyte parameters and hemoglobin F production in sickle cell disease patients undergoing hydroxyurea therapy. AB - Hemoglobin F (HbF) is an effective inhibitor of HbS polymerization. Hydroxyurea (HU) is used to increase HbF synthesis and improve the clinical course of sickle cell disease (SCD) patients. We studied a series of laboratory parameters concerning HbF production and reticulocyte response, and compared data between two groups: 1) 13 SCD patients treated with HU, and 2) 33 untreated SCD patients. Higher values of Hb concentration, mean cell volume (MCV), mean cell hemoglobin (MCH), mean reticulocyte volume (MRV), HbF concentration, percentage of F-cells, and amount of HbF/F-cells were observed in the treated group of patients. There was no correlation between Hb and HbF elevations. The reticulocyte count, immature reticulocyte count, mean fluorescence index (MFI), and neutrophil count were significantly lower in treated patients. Taken together, these findings suggest that a decreased hemolytic process occurred in patients undergoing HU treatment. There was a significant correlation between MCV and HbF, between MRV and HbF, and between MRV and F-cell in patients taking HU. These data indicate that macroreticulocytes correspond to F-reticulocytes, and that an increase in MRV in SCD patients using HU may be an indirect signal of F-cell production. The concentration of HbF/F-cells was higher in patients treated with HU, but this increase apparently was independent of F-cell production. Reticulocyte (RTC) parameters, as assessed by hematological analyzers, may be useful for following erythropoietic changes in patients receiving HU, and can indirectly indicate HbF and F-cell production induced by HU therapy. PMID- 12640634 TI - Copper carbene complexes: advanced catalysts, new insights. AB - Controlling the selectivity of reactive intermediates is a major goal of organic chemists. Carbenes generated by thermal or photochemical extrusion of nitrogen from diazo compounds defy external control and tend to give complex product mixtures. However, the catalyzed decomposition of diazo compounds gives rise to highly selective "carbenoids" which have found extensive application in synthesis although little is known of their precise nature. On the other hand, a large variety of carbene metal complexes have been prepared and characterized which more or less lack carbenoid reactivity. Structural evidence for the transients of catalyzed diazo decomposition is becoming available through experiment and computation. Although the present report focuses on copper complexes, outstanding results with other metals are also covered. PMID- 12640633 TI - Chemical genetics goes (zebra) fishing. PMID- 12640635 TI - meso Compounds: stepchildren or favored children of stereoselective synthesis? AB - After more than a quarter of a century of development, the methodology of stereoselective synthesis appears to be fully matured. In line with this, the potential that meso compounds offer in stereoselective synthesis is clearly recognized. The use of meso compounds in synthesis is, however, in no way commensurate with this potential, because, ironically, the synthesis of meso compounds in the first place is a problem of stereoselective synthesis. Present day methodology does not provide many useful solutions to this problem. This Review therefore addresses the strategies available for the synthesis of more elaborate meso compounds whose stereogenic centers have a distance >1,4 between them. meso Compounds with more than four stereogenic centers are also considered. The criteria used in choosing from several strategies in the synthesis of such compounds are discussed. PMID- 12640636 TI - New tricks for an old dog: the carbonate ion as a building block for networks including examples of composition [Cu6(CO3)12(C(NH2)3)8]4- with the sodalite topology. PMID- 12640637 TI - Time-controlled desorption of ultrathin polymer films triggered by enzymatic degradation. PMID- 12640638 TI - Recognition of stereoregular polymers by using structurally regulated ultrathin polymer films. PMID- 12640639 TI - A new route to organic nanotubes from porphyrin dendrimers. PMID- 12640640 TI - Diastereospecific photochemical dimerization of a stilbene-containing daisy chain monomer in solution as well as in the solid state. PMID- 12640641 TI - Aluminum dihydroxide with terminal OH groups: an unprecedented congener of boronic acid. PMID- 12640643 TI - Direct synthesis of propylene oxide with CO2 as the solvent. PMID- 12640642 TI - Combining SPOT synthesis and native peptide ligation to create large arrays of WW protein domains. PMID- 12640644 TI - [(tBu2MeSi)3Ge]+: an isolable free germyl cation lacking conjugation to pi bonds. PMID- 12640645 TI - Overcoming the "upper bound" in polymeric gas-separation membranes. PMID- 12640646 TI - The role of Mo atoms in nitrogen fixation: balancing substrate reduction and dihydrogen production. PMID- 12640647 TI - High-throughput catalytic science: parallel analysis of transients in catalytic reactions. PMID- 12640648 TI - A zeolite structure (ITQ-13) with three sets of medium-pore crossing channels formed by 9- and 10-rings. PMID- 12640649 TI - Oligo(2,6-anthrylene)s: acene-oligomer approach for organic field-effect transistors. PMID- 12640650 TI - Synthesis and structural characterization of redox-active dodecamethoxoheptaoxohexavanadium clusters. PMID- 12640651 TI - Solid-phase-assisted solution-phase synthesis with minimum purification preparation of 2-deoxyglycoconjugates from thioglycosides. PMID- 12640652 TI - Synthesis of a chiral tube. PMID- 12640653 TI - Picotube tetraanion: a novel lithiated tubular system. PMID- 12640655 TI - Sex hormone-induced alterations in the activities of antioxidant enzymes and lipid peroxidation status in the prostate of Noble rats. AB - BACKGROUND: Oxidative stress has been implicated in prostate carcinogenesis. In the Noble rat model, treatment of rats with testosterone (T) plus 17beta estradiol (E(2)) induced dysplasia and adenocarcinoma. Previous reports from us and other have indicated a linkage between steroid hormones and oxidative status in prostate cells in vivo and in vitro. Here, we provide further evidence that androgens and estrogens could induce a lobe-specific shift of prooxidant antioxidant balance and alterations in antioxidant enzyme activities, leading to oxidative stress in the rat prostate gland in vivo. METHODS: Male Noble rats were subjected to single (T, E(2), or diethylstilbestrol [DES] alone) or combined (T + E(2) or T + DES) hormone treatments. Lipid peroxidation status and antioxidant enzyme activities of superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), glutathione reductase (GR), glucose-6-phosphate dehydrogenase (G6PD) were assayed spectrophotometrically. RESULTS: Treatment of rats with T did not alter lipid peroxidation in either the ventral prostate (VP) or the dorsolateral prostate (DLP). In contrast, exposure of rats to DES or E(2) modestly elevated lipid peroxidation in rat VP or DLP, respectively. Of importance, T + DES and T + E(2) treatments of rats induced marked increases in lipid peroxidation in the VP and the DLP, respectively. Antioxidant enzyme activities in the VP and DLP exhibited differential responses to sex hormone challenges. The activities of catalase, GPx, GR, and G6PD were mostly suppressed in either single or dual hormone-treated DLP, whereas there is a general increase of GR and G6PD activities in the VP after hormonal exposures. The changes in T + DES-treated VP were most dramatic with a marked activation of GPx (by one-fold), GR (by one fold), and G6PD (by five-fold). CONCLUSION: The lobe-specific differential responses of hormone-induced oxidative stress and modulations of antioxidant enzymatic defenses in the rat prostate suggest that reactive oxygen species may play a role in hormone-induced prostate carcinogenesis. PMID- 12640656 TI - Polarity of prostate specific membrane antigen, prostate stem cell antigen, and prostate specific antigen in prostate tissue and in a cultured epithelial cell line. AB - BACKGROUND: Madin-Darby canine kidney (MDCK) cells are immortalized epithelial cells that have been used extensively as a model system to study intracellular molecular trafficking, polarized expression, and secretion of proteins in various epithelia. In order to determine if MDCK cells might serve as a model to study molecular events within prostate epithelial cells, we have evaluated the polarized distribution of three prostate restricted proteins, PSMA, PSCA, and PSA, in situ, and in MDCK cells. METHODS: Using immunofluorescence, confocal microscopy, cell surface biotinylation, antibody internalization, and biochemical assays we evaluated surface expression and secretion of three prostate restricted proteins expressed in MDCK cells. We compared these patterns of expression to results observed within prostatic epithelium. RESULTS: We demonstrate that PSMA is localized primarily to the apical plasma membrane in both the prostatic epithelium and transfected MDCK cells, whereas PSCA is expressed in a non polarized fashion. We also show that PSA is secreted predominantly from the apical surface of transfected MDCK cells, consistent with in vivo observations. CONCLUSIONS: Similar patterns of localization among MDCK and prostatic epithelial cells suggest that the mechanisms of polarized sorting within these cell types are conserved. Thus, MDCK cells offer a useful model system to study mechanisms of targeting of these proteins within the prostate. PMID- 12640657 TI - Tissue microarray analysis reveals prognostic significance of syndecan-1 expression in prostate cancer. AB - BACKGROUND: Tissue microarrays (TMA) have recently emerged as powerful tools to rapidly analyze the clinical significance of new molecular markers in human tumors. Here, we have tested several molecular markers on a prostate TMA containing 637 different specimens. METHODS: The specimens were from 551 patients with prostate cancer and long-term follow-up information on progression (median 5.3 years), tumor-specific and overall survival (median 5.9 years). Eighty-six specimens from benign prostatic hyperplasia were included as controls. Expression of Ki67, Bcl-2, p53, CD-10 (neutral endopeptidase), and syndecan-1 (CD-138) was analyzed by immunohistochemistry. RESULTS: Gleason grade and Ki67 Labeling Index (LI) were independent predictors of early recurrence and poor survival. Bcl-2 predicted early recurrence, whereas p53 was associated with poor survival. Syndecan-1 overexpression also predicted early recurrence and was significantly associated with tumor specific survival, high Gleason grade, Ki67 LI, and Bcl-2 overexpression. Neoadjuvant hormonal therapy was associated with overexpression of Bcl-2 and inhibition of Ki67 LI and CD-10, but did not affect the expression of the remaining markers. CONCLUSIONS: The results of this TMA study confirm a dominant prognostic significance of Gleason grading and Ki67 LI in prostate cancer, as compared to a less pronounced role of Bcl-2, and p53. We identified syndecan-1 as a new prognostic factor and provide evidence for an androgen dependent regulation of CD-10 expression. PMID- 12640658 TI - Cellular senescence in the pathogenesis of benign prostatic hyperplasia. AB - BACKGROUND: Senescent cells accumulate in tissues with age and show changes in protein expression that may influence the function of adjacent cells and contribute to the development of tissue pathologies associated with aging. Benign prostatic hyperplasia (BPH) is an extremely common disease of older men characterized by increased growth of prostatic epithelial and stromal cells. In BPH, there is an increased expression of Il-1alpha by prostatic epithelial cells that results in elevated expression of FGF7 by stromal cells, which in turn is strongly correlated with epithelial proliferation. METHODS: Human BPH tissue and primary cultures of prostatic epithelial cells were analyzed by histochemical and quantitative assays for senescence-associated beta galactosidase (SA-beta gal). Il-1alpha expression was localized by immunohistochemistry and Il-1alpha tissue content determined by enzyme-linked immunoabsorption assay. RESULTS: Expression of Il-1alpha is significantly increased in vitro when cultured prostatic epithelial cells undergo senescence. In BPH tissue a substantial population of epithelial cells express senescence-associated beta galactosidase (SA-beta gal), a marker of cellular senescence. By quantitative assay, SA-beta gal activity is correlated with both tissue levels of Il-1alpha and the severity of BPH. CONCLUSIONS: One mechanism driving BPH in older men is the accumulation of senescent epithelial cells expressing Il-1alpha, which in turn increases FGF7 secretion and proliferation of non-senescent epithelial cells. Thus there is a mechanistic linkage between cellular senescence and one of the most common pathologies of older men. PMID- 12640659 TI - MicroPET imaging of prostate cancer in LNCAP-SR39TK-GFP mouse xenografts. AB - BACKGROUND: The aim of this study was to develop models that allow serial, noninvasive imaging of human prostate cancer cells in immunodeficient mice using a dedicated small animal positron emission tomography scanner (microPET). METHODS: LNCaP tumor cells were stably transduced ex-vivo with the mutant herpes simplex virus type 1 thymidine kinase (HSV-sr39tk) PET reporter gene and green fluorescent protein (GFP). The stably transduced LNCaP cells were then enriched via fluorescent cell sorting and implanted into SCID mice. Beginning 2 weeks after tumor cell inoculation, mice were repeatedly scanned by microPET performed 1 hr after tail-vein injection of approximately 200 muCi Fluorine-18 labeled penciclovir ((18)F-FHBG). PET-images were correlated to tumor size, % injected dose (ID)/g tumor tissue, PSA levels, autoradiography, and histology. RESULTS: Monitoring LNCaP xenografts using microPET and our reporter gene approaches is feasible. MicroPET was capable of detecting subcutaneous tumors as small as 3 mm in diameter (approximately 0.2% ID/g). The magnitude of (18)F-FHBG-uptake in PET images correlated with the tumor volumes and the serum PSA levels. Other non-HSV1 TK-specific tracers were also studied. While (18)F-flurodeoxyglucose ((18)F-FDG) gave poor imaging results in LNCaP cells, (11)C-acetate gave satisfactory images. CONCLUSIONS: We demonstrated the feasibility of monitoring prostate cancer xenografts in a mouse model using microPET and the HSV1-sr39tk PET reporter gene/(18)F-FHBG reporter probe system. Extension of this approach may allow repetitive imaging of tumor metastases. PMID- 12640660 TI - Biochemical recurrence following radical prostatectomy: a comparison between prostate cancers located in different anatomical zones. AB - BACKGROUND: To assess whether differences of biochemical recurrence after radical prostatectomy exist between prostate cancers located in the transition zone (TZ) and peripheral zone (PZ). METHODS: The 5-year biochemical recurrence rate of 307 patients was evaluated. A serum prostate specific antigen (PSA) level > or =0.1 ng/ml was defined as biochemical failure. Cancers were characterized by the location of the largest tumor area as TZ or PZ cancers. Pure PZ cancers were matched to TZ cancers by comparable pathological tumor stage, Gleason score, and surgical margin status. RESULTS: In 63 (20.5%) patients the largest tumor area was located in the TZ. A Kaplan-Meier analysis of the matched pairs calculated an 80% actuarial cure rate of TZ cancers compared to 89% of pure PZ cancers (log rank test P = 0.742). CONCLUSIONS: TZ and pure PZ cancers matched by comparable pathological tumor stage, Gleason score, and surgical margin status showed no statistical difference in regard to biochemical cure following radical prostatectomy. PMID- 12640661 TI - Molecular characterization of prostatic small-cell neuroendocrine carcinoma. AB - OBJECTIVES: A subset of prostate carcinomas is composed predominantly, even exclusively, of neuroendocrine (NE) cells. In this report, we sought to characterize the gene expression profile of a prostate small cell NE carcinoma by assessing the diversity and abundance of transcripts in the LuCaP 49 prostate small cell carcinoma xenograft. METHODS: We constructed a cDNA library (PRCA3) from the LuCap 49 prostate small cell xenograft. Single pass DNA sequencing of randomly selected cDNA clones followed by sequence assembly and annotation produced a library of Expressed Sequence Tags (ESTs) representing the LuCaP 49 transcriptome. Comparative sequence analysis with ESTs derived from prostate adenocarcinoma libraries was performed using statistical algorithms designed to identify differentially expressed sequences. Putative NE cell-specific genes were further examined by Northern analysis. RESULTS: Sequence assembly and analysis identified 1,447 distinct genes expressed in the LuCaP 49 cDNA library. These include cDNAs encoding the NE markers secretogranin (SCG2), CD24, and ENO2. Northern analysis revealed that three additional genes, ASCL1, INA, and SV2B are expressed in LuCaP 49 but not in various prostate cancer cell lines or xenografts. Fifteen genes were identified with a statistical probability (P > 0.9) of being up-regulated in LuCaP 49 small cell carcinoma relative to prostate adenocarcinoma (two primary prostate adenocarcinomas and the LNCaP prostate adenocarcinoma cell line). CONCLUSIONS: Prostate small cell carcinoma expresses a diverse repertoire of genes that reflect characteristics of their NE cell of origin. ASCL1, INA, and SV2B are potential molecular markers for small cell NE tumors and NE cells of the prostate. This small cell NE carcinoma gene expression profile may yield insights into the development, progression, and treatment of subtypes of prostate cancer. PMID- 12640662 TI - Laminin alpha-1, alpha-3, and alpha-5 chain expression in human prepubertal [correction of prepubetal] benign prostate glands and adult benign and malignant prostate glands. AB - BACKGROUND: Laminins (Lns) are a family of extracellular matrix glycoproteins located in the basement membrane (BM) of epithelial cells. They exist as heterotrimers composed of an alpha, beta, and gamma chain. Presently, five alpha (alpha1-5), three beta (beta1-3), and three gamma (gamma1-3) chains have been identified with different combinations of these chains resulting in 14 laminin heterotrimers thus far identified (1, 3-5). METHODS: In this study, using immunohistochemistry with chain-specific antibodies, we characterized the expression of the alpha1 (Lns-1/3), alpha3 (Lns 5,6,7), and alpha5 (Lns 10/11) chains in fetal, newborn, infant, prepubertal, and adult benign and malignant prostate glands. RESULTS: In general, alpha1 expression was higher in normal fetal prostate glands and declined by full-term birth, whereas the alpha3 and alpha5 chains remained highly expressed in the adult normal glands. In carcinoma alpha1 (Lns 1/3) and alpha5 (Lns 5,6,7) are lost, whereas alpha5 (Lns 10/11) persists. CONCLUSIONS: Alpha 1 (Lns 1/3) is prominent in BM, but is replaced by a laminin matrix rich in alpha3 (Lns 5,6,7) and alpha5 (Lns 10/11) in benign adult prostate glands. In carcinoma, both alpha1 (Lns-1/3) and alpha3 (Lns 5,6,7) are not expressed with persistence of a BM rich in alpha5 (Lns 10/11). PMID- 12640663 TI - Loss of CD10 (neutral endopeptidase) is a frequent and early event in human prostate cancer. AB - BACKGROUND: We hypothesized that the aggressive LNCaP-derived androgen independent cell line, CL1, might differ from LNCaP in their repertoire of cell surface markers and that these differences might typify changes that occur during clinical prostate cancer progression. METHODS: The cell surface marker expression profiles of CL1 and LNCaP were examined using flow cytometry. Markedly differential gene expression was confirmed using RT-PCR and further examined using immunohistochemistry among the prostate cancer cell lines LAPC-4, LNCaP, CL1, CL2, DU145, and PC-3. The expression of the most markedly differentially expressed surface marker, CD10, was further explored in a tissue microarray containing radical prostatectomy samples from 219 hormone naive prostate cancer patients. RESULTS: There were marked differences in the expression of CD10, CD13, CD26, CD33, CD44, CD54, CD55, and CD104 between CL1 and LNCaP. Results from both the RT-PCR and immunohistochemistry confirmed the differential expression and found that CD10 demonstrated a pattern of expression in hormone sensitive but not hormone refractory cell lines. When CD10 expression was examined in a tissue microarray, CD10 expression was below the 25th percentile of matched normal prostate tissue in 68% of prostate cancers, below the median expression of matched normal prostate tissue in 86% of cancers, and completely absent in 34% of cancers. Samples of prostatic intraepithelial neoplasia demonstrated CD10 expression that was intermediate between normal prostatic tissue and prostate cancer. Among prostate cancer patients, CD10 expression did not correlate with Gleason score, pathological stage, or biochemical recurrence following radical prostatectomy. CONCLUSIONS: These findings demonstrate that loss or decreased expression of CD10 is an early and frequent event in human prostate cancer and implicates CD10 as a potential therapeutic target for early stage hormone sensitive prostate cancer. PMID- 12640665 TI - Internal connectivity of the homing pigeon (Columba livia) hippocampal formation: an anterograde and retrograde tracer study. AB - The avian hippocampal formation (HF) is a structure necessary for learning and remembering aspects of environmental space. Therefore, understanding the connections between different HF regions is important for determining how spatial learning processes are organized within the avian brain. The prevailing feed forward, trisynaptic internal connectivity of the mammalian hippocampus and its importance for cognition have been well described, but the internal connectivity of the avian HF has only recently been investigated. To examine further the connectivity within the avian HF, small amounts of cholera toxin subunit B, primarily a retrograde tracer (n = 15), or biotinylated dextran amine, primarily an anterograde tracer (n = 10), were injected into localized regions of the HF. Examination of the immunohistochemically labeled tissue showed projections from extrinsic sensory processing areas into dorsolateral HF and the dorsal portion of the dorsomedial HF (DMd). DMd in turn projected into the medial (VM) and lateral (VL) ventral cell layers. A projection from VM into VL was found, and together these areas and DM provided input into the contralateral ventral cell layers. Ipsilaterally, a ventral portion of dorsomedial HF (DMv) received input from VL and VM. From DMv, projections exited HF laterally. The highlighted projections formed a discernible feed-forward processing network through the avian HF that resembled the trisynaptic circuit of the mammalian HF. PMID- 12640664 TI - Comparative expression patterns of T-, N-, E-cadherins, beta-catenin, and polysialic acid neural cell adhesion molecule in rat cochlea during development: implications for the nature of Kolliker's organ. AB - We investigated the expression patterns of several cell adhesion molecules (CAMs) during rat cochlea ontogeny, from embryo day 16 to adulthood, with the use of immunohistochemistry: neural cadherin (N-cad) and polysialic acid neural CAM (PSA NCAM) as two different neural CAM paradigms; epithelial cadherin (E-cad), which was restricted to the epitheloid phenotype; and the cytoplasmic domain-free truncated-cadherin (T-cad). We made the following observations. (1) T-cad was present in all types of fibrocyte and in subdomains within the pillar cells. (2) E- and N-cad were expressed with mutually exclusive patterns and did not overlap with T-cad. All cochlear epithelial cells, including the sensory outer hair cells (OHCs), were E-cad-positive, except for the negative inner hair cells (IHCs) and the nonsensory Kolliker's organ domain close to the IHCs. N-cad expression appeared first in the developing IHCs and then in the neighboring Kolliker's organ in an increasingly mediolateral gradient in opposition to the E-cad gradient. The OHCs, which are never N-cad positive, intensively expressed E-cad, as did the Hensen cells at the beginning of their differentiation. (3) The cadherin-linked molecule beta-catenin, absent in fibrocytes, was detected in all epithelial cell membranes and was prominent in the E-cad-rich modiolar extremity of Kolliker's organ. (4) Gradual PSA-NCAM expression was observed in the lateral portion of Kolliker's organ, and the intense PSA-NCAM expression was seen surrounding the IHCs. As development proceeded, PSA-NCAM immunoreactivity progressively became restricted to the basal poles of the IHCs, where it remained in the adult rat cochlea, suggesting a synaptic plasticity. Synaptic plasticity in rat cochlea and hypotheses about T-cad functions and neosensory features of the Kolliker's organ are discussed. PMID- 12640667 TI - Retinal influences specify cortico-cortical maps by postnatal day six in rats and mice. AB - Studies of callosal projections in striate cortex show that the retina is involved in the development of topographical connections. In normal animals callosal fibers connect retinotopically corresponding, nonmirror-symmetric cortical loci, whereas in animals bilaterally enucleated at birth, callosal fibers connect topographically mismatched, mirror-symmetric loci. Moreover, in rodents the overall pattern of visual callosal connections is adult-like by postnatal day 12 (P12). In this study we delayed the onset of retinal deafferentation in rats and mice in order to determine the period when retinal influences are critically needed for the development of retinotopically matched callosal linkages. Callosal maps were revealed by placing small injections of retrogradely and anterogradely transported tracers into different loci of lateral striate cortex. We found that the patterns of callosal linkages in rats enucleated at P12, P8, and P6 were nonmirror-symmetric, as in normally reared rats. In contrast, the patterns of linkages in rats enucleated at P4 closely resembled the mirror-symmetric pattern seen in rats enucleated at birth (P0). A similar reversal in topography (from symmetric to nonsymmetric) occurred in mice when enucleation was delayed from P4 to P6. These findings indicate that retinal input prior to P6, but not prior to P4, is sufficient for specifying normal callosal topography. Moreover, they suggest that development of retinotopically matched callosal linkages depends critically on retinal influences during a brief period between P4 and P6, when callosal connections are still very immature. PMID- 12640666 TI - Projections from the paraventricular nucleus of the thalamus to the rat prefrontal cortex and nucleus accumbens shell: ultrastructural characteristics and spatial relationships with dopamine afferents. AB - The paraventricular nucleus of the thalamus (PVT) participates in the functional integration of limbic cortical and striatal circuitry. In the rat, the PVT projects to the deep layers of the medial prefrontal cortex (PFC) and to the shell of the nucleus accumbens (NAc). However, the synaptic organization of PVT afferents within these regions remains undescribed. Furthermore, although dopamine (DA) modulates excitatory glutamate transmission in both areas, possible anatomic substrates for specific DA modulation of PVT inputs have not yet been investigated. To address these issues, immunoperoxidase labeling for tyrosine hydroxylase (TH) in DA axons was combined with anterograde tract-tracing, either by biotinylated dextran amine (BDA) labeled with immunogold-silver or by degeneration after lesions of the PVT. In both regions, and with either tracing method, PVT terminals formed primarily asymmetric axospinous synapses; in the NAc, a proportion of PVT terminals also synapsed onto dendrites. PVT profiles in both regions were often seen in direct apposition to TH-immunoreactive axons; this association was more evident in the NAc where the DA innervation is denser. Within the PFC, PVT profiles and TH-labeled axons were occasionally apposed to the same dendrites, but synaptic specializations were not typically seen at these seeming points of convergence. Within the NAc, PVT profiles occasionally made synapses onto spines and distal dendrites that received convergent synapses from TH-immunoreactive varicosities. These findings represent the first demonstration of postsynaptic convergence between DA and thalamic afferents to a striatal region and are consistent with direct synaptic modulation of PVT transmission by DA in the NAc but not the PFC. PMID- 12640668 TI - Tactile sensory function in the forearm of the monotreme Tachyglossus aculeatus. AB - Peripheral tactile neural mechanisms in the forepaw of the echidna (Tachyglossus aculeatus, from the order Monotremata) were investigated to establish the extent of correspondence or divergence that has emerged over the widely different evolutionary paths taken by monotreme and placental mammals. Electrophysiological recordings were made in anesthetized echidnas from 29 single tactile sensory nerve fibers isolated in fine strands of the median or ulnar nerves of the forearm. Controlled tactile stimuli were applied to the forepaw glabrous skin to classify fibers, initially, into two broad divisions, according to their responses to static skin displacement. One displayed slowly adapting (SA) response properties, and the other showed a selective sensitivity to the dynamic components of the skin displacement. The SA class was made up of low-threshold SA fibers and other less sensitive SA fibers, and the purely dynamically sensitive tactile fibers could be subdivided according to vibrotactile sensitivity and receptive field characteristics into a rapidly adapting (RA) class, sensitive to low-frequency (< or =50-Hz) vibration, that resembled a corresponding RA class in placental species, and another class, sensitive to a broader range of vibrotactile frequencies (approximately 50-300 Hz), that may represent a monotreme equivalent of the Pacinian corpuscle (PC)-related fiber class of placental mammals. The differential tactile sensitivity of the three principal fiber classes and their individual coding characteristics, determined by quantitative stimulus-response analysis, indicate, first, that this triad of fiber classes can subserve high-acuity tactile signalling from the echidna footpad and, second, that peripheral tactile sensory mechanisms are highly conserved across evolutionarily divergent mammalian orders. PMID- 12640669 TI - Visually guided injection of identified reticulospinal neurons in zebrafish: a survey of spinal arborization patterns. AB - We report here the pattern of axonal branching for 11 descending cell types in the larval brainstem; eight of these cell types are individually identified neurons. Large numbers of brainstem neurons were retrogradely labeled in living larvae by injecting Texas-red dextran into caudal spinal cord. Subsequently, in each larva a single identified cell was injected in vivo with Alexa 488 dextran, using fluorescence microscopy to guide the injection pipette to the targeted cell. The filling of cells via pressure pulses revealed distinct and often extensive spinal axon collaterals for the different cell types. Previous fills of the Mauthner cell had revealed short, knob-like collaterals. In contrast, the two segmental homologs of the Mauthner cell, cells MiD2cm and MiD3cm, showed axon collaterals with extensive arbors recurring at regular intervals along nearly the full extent of spinal cord. Furthermore, the collaterals of MiD2cm crossed the midline at frequent intervals, yielding bilateral arbors that ran in the rostral caudal direction. Other medullary reticulospinal cells, as well as cells of the nucleus of the medial longitudinal fasciculus (nMLF), also exhibited extensive spinal collaterals, although the patterns differed for each cell type. For example, nMLF cells had extensive collaterals in caudal medulla and far-rostral spinal cord, but these collaterals became sparse more caudally. Two cell types (CaD and RoL1) showed arbors projecting ventrally from a dorsally situated stem axon. Additional cell-specific features that seemed likely to be of physiological significance were observed. The rostral-caudal distribution of axon collaterals was of particular interest because of its implications for the descending control of the larva's locomotive repertoire. Because the same individual cell types can be identified from fish to fish, these anatomical observations can be directly linked to data obtained in other kinds of experiments. For example, 9 of the 11 cell types examined here have been shown to be active during escape behaviors. PMID- 12640670 TI - Hippocampal cell genesis does not correlate with spatial learning ability in aged rats. AB - Aging in rodents is known to lead to deficits in spatial learning and memory, including decreased performance on the Morris water maze. Recent attention has focused on the possible role of adult hippocampal neurogenesis in regulating spatial learning and memory. Therefore, in this study, we have examined levels of hippocampal cell proliferation in relation to water maze performance in aged and young male Fischer 344 rats. Aged rats (24 months old) were divided into aged unimpaired and aged-impaired groups based on comparison with performance of young animals. Animals received five daily injections of the thymidine-analog bromodeoxyuridine (BrdU) and were killed 1 week later. Total numbers of BrdU labeled cells were quantified in the hippocampal dentate gyrus and hilus and were related to behavioral performance. Whereas aging was associated with a significant reduction in the number of BrdU-labeled cells, behavioral impairment with aging was not associated with a further reduction in BrdU labeling. In the context of aging, these finding do not support a direct relationship of adult hippocampal neurogenesis with learning and memory capability. PMID- 12640671 TI - Circulating nucleosomes and response to chemotherapy: an in vitro, in vivo and clinical study on cervical cancer patients. AB - It is known that cell-free DNA circulates in plasma/serum of patients with cancer and that part of this DNA circulates as nucleosomes that can be quantified by ELISA. We analyzed the effect of tumor and chemotherapy upon the levels of nucleosomes in vitro, in vivo and in cervical cancer patients. The levels of nucleosomes pre- and post-treatment were correlated with response in 11 patients receiving chemotherapy. Nucleosomes were determined in nude mice treated with or without cisplatin and carrying tumors generated with HeLa cells, and in the cell lysate and supernatant of HeLa cells exposed to cisplatin in culture. In addition, nucleosomes were determined at different time points in patients and in rats receiving chemotherapy. Nucleosomes were higher in patients that controls (1,760 vs. 601, p = 0.0001). After 24 hr of treatment with oxaliplatin and gemcitabine, the levels decreased in 6 patients of whom 5 had response. Nucleosome levels differed between mice xenografted and not xenografted (765 vs. 378, p = 0.001) and between xenografted treated with or without cisplatin (650 vs. 765, p = 0.010), but not in tumor-free animals treated and untreated with cisplatin (378 vs. 379, p = 0.99). In vitro, nucleosomes reached at peak 8 hr in cell lysates to decrease thereafter, whereas in supernatant, levels continued to increase up to 24 hr. Serial determination of nucleosomes in patients showed a rise within 6-12 hr and then a reduction to below the basal at 24 hr. In rats, nucleosomes had no major changes in those receiving oxaliplatin or the triple combination of cisplatin, gemcitabine and paclitaxel as compared to untreated controls. An overdose of this triple combination produced a transient elevation of almost 1,000 AU over the basal. Our results demonstrate that most of circulating nucleosomes originate from the tumor and that chemotherapy produces an early rise most likely due to tumor apoptosis and that nucleosomes are rapidly cleared from circulation. On the contrary, chemotherapy within the therapeutic range of doses has no effect on nucleosome levels in healthy mice and rats. This data suggests that the determination of circulating nucleosomes pre- and post treatment could be a useful test to predict response to chemotherapy in cancer patients. PMID- 12640672 TI - Metabolic abnormalities (hypertension, hyperglycemia and overweight), lifestyle (high energy intake and physical inactivity) and endometrial cancer risk in a Norwegian cohort. AB - Since high energy intake, inactivity, hypertension and diabetes are linked to obesity and an unfavorable hormonal profile, we wanted to test whether energy intake, physical activity, blood pressure and serum glucose are related to the risk of endometrial cancer independent of the body mass index (BMI). A cohort of 24,460 women, aged 20-49 years, attended a Norwegian health screening twice during 1974-1981; they answered questions about diet, physical activity and chronic diseases, and their height, weight, blood pressure and non-fasting serum glucose were measured. By the end of 1996, during 15.7 years of follow-up, 130 cases of endometrial carcinomas were identified. The relative risks (RRs) for endometrial cancer were estimated in proportional hazards models including potentially confounding factors. Obese women (BMI > or = 30 kg/m(2)) were at 2.6 times increased risk of endometrial cancer compared to normal weight women (BMI < 25 kg/m(2)) (RR = 2.57, 95%CI = 1.61-4.10). Among overweight women (BMI > or = 25 kg/m(2)), non-fasting serum glucose in the upper quartile vs. in the lower quartile was associated with a 2.4 times increase in risk (RR = 2.41, 95%CI = 1.08-5.37), whereas among obese women, blood pressure above 140/90 mmHg vs. below 140/90 mmHg in both surveys was associated with a 3.5 times increase in risk (RR = 3.47, 95%CI = 1.24-9.70). Especially in women younger than 50 years, high energy intake (5,044-6,401 kJ/day) conferred higher risk compared to low energy intake (< 4266 kJ/day) (RR = 3.40, 95%CI = 1.52-7.60). Increasing recreational activity tended to be protective. Among obese women with non-sedentary jobs at both screenings, RR declined to 0.18 (95%CI = 0.05-0.62) as the level of sustained occupational activity increased (p(trend) = 0.03). Our results suggest that inactivity and high energy intake are major risk factors for endometrial cancer independent of BMI, and that hypertension and relative hyperglycemia are significant markers of risk, especially among the heaviest women. PMID- 12640673 TI - Overexpression of Id-1 is associated with poor clinical outcome in node negative breast cancer. AB - Id-1 is an important regulator of cellular growth and differentiation and controls malignant progression of breast cancer cells. The aim of our study was to assess the clinical impact of Id-1 expression in breast cancer, i.e., its potential impact on prognosis and prediction of treatment response. Id-1 protein expression was determined immunohistochemically in 191 patients with lymph-node negative breast cancer, and univariate and multivariate survival analysis was carried out. Fifteen (7.9%) specimens showed strong expression, 75 (39.3%) moderate, 55 (28.8%) weak expression and 46 (24.1%) cases no expression of Id-1. Patients with strong or moderate Id-1 expression had a significant shorter overall (p = 0.003, Cox regression) and disease-free survival (p = 0.01, Cox regression) compared to those with absent or low expression. Progesterone receptor density was significantly higher in breast cancers with absent/low Id-1 expression compared to those with moderate/strong expression (p < 0.001, t-test). Id-1 expression was significantly stronger in cases positive for p16(INK4a) expression compared to those negative for p16 (p = 0.049, Mann-Whitney test). The influence of Id-1 on clinical outcome seems much stronger in patients with negative estrogen receptor status compared to those with positive status, who received receptor antagonists as adjuvant therapy in most cases. Overexpression of Id-1 protein represents a strong independent prognostic marker in node negative breast cancer, and future therapies inhibiting Id-1 expression might be beneficial for these patients. Our results also suggest that due to the apparent interaction of Id-1 with the steroid-receptor system in breast cancer, hormonal therapies might influence Id-1 expression and its impact on clinical outcome. PMID- 12640674 TI - Requirement of both mucins and proteoglycans in cell-cell dissociation and invasiveness of colon carcinoma HT-29 cells. AB - Human colon carcinomas are characterized by an aberrant expression of mucins, which in some case leads to an abundant presence of mucus such as in mucinous and signet ring cell carcinomas. Cellular cloning of the human colon carcinoma cell line HT-29 (HT-29 STD), which is mainly composed of undifferentiated cells, yielded a highly mucin-secreting variant (HT-29 5M21). The latter cloned cells cultured on plastic display a polarized organization with an apical secretion of MUC5AC mucin (Lesuffleur et al., Int J Cancer 1998;76:383-92.). Our aim was to study these 2 cell-types as for the invasive and adhesive properties with regard to the function of E-cadherin. HT-29 STD cells were noninvasive in collagen type I, whereas HT-29 5M21 cells were invasive, and the latter behavior was connected to a loss of function of E-cadherin. Likewise, HT-29 5M21 cells were characterized by a cell-cell adhesion independent of E-cadherin, in contrast to the E-cadherin dependent cell-cell adhesion of HT-29 STD cells. Immunofluorescence of HT-29 5M21 cells cultured on collagen type I showed the disappearance of the polarized organization, with a redistribution of apical mucins to the entire cell surface. Treatment of HT-29 5M21 cells by 1-benzyl-2 acetamido-2-deoxy-alpha-D-galactopyranoside (GalNAcalpha-O-bn) or by beta-D xyloside revealed that both mucins and proteoglycans were involved in the loss of E-cadherin function. The use of specific antibodies allowed to show that MUC5AC, MUC1 and heparan sulfate proteoglycans cooperated in the formation of a biological inhibitory complex towards the function of E-cadherin in this invasive HT-29 clone. PMID- 12640675 TI - BCL-2/JH translocation in peripheral blood lymphocytes of unexposed individuals: lack of seasonal variations in frequency and molecular features. AB - BCL-2/J(H) rearrangement has been proposed as a biomarker for evaluating the genotoxicity of occupational and environmental exposures. Available data on time related modification of this rearrangement in peripheral blood lymphocytes in unexposed healthy individuals is scarce. We investigated the characteristics of BCL-2/J(H) rearrangements in 33 adults unexposed to genotoxins at 2 seasonal time points: winter and spring. BCL-2/J(H) rearrangement was detected in 79% of individuals (detection limit = 8.48 x 10(-8)). Its frequency ranged from <1 to 40 translocations per million lymphocytes with a significant (p = 0.04) positive correlation with age. No significant modifications of BCL-2/J(H) rearrangement frequency or in the number of clones harboring this rearrangement were observed according the 2 time points. No obvious influence of season-related environmental factors on frequency or molecular features of BCL-2/J(H) rearrangements was found in this population suggesting that this would not be a confounding factor. PMID- 12640676 TI - Profile of gene expression induced by the tumour promotor TPA in murine epithelial cells. AB - Malignant transformation of mouse skin by chemical carcinogens and tumour promoters, such as the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), is a multistage process that leads to squamous cell carcinoma (SCC) formation. In an effort to identify tumour-associated genes, we studied the influence of short term TPA-treatment on the gene expression profile of murine skin. A comprehensive microarray with some 5,000 murine gene specific cDNA fragments was established and hybridised with pooled RNA derived from control and TPA-treated dorsal skin samples. Of these genes, 54 were up- and 35 were down-regulated upon TPA application. Additionally, we performed suppression subtractive hybridisation (SSH) with respective RNA pools to generate and analyse a cDNA library enriched for TPA-inducible genes. Expression data of selected genes were confirmed by quantitative real-time PCR and Northern blot analysis. Comparison of microarray and SSH data revealed that 26% of up-regulated genes identified by expression profiling matched with those present in the SSH library. Besides numerous known genes, we identified a large set of unknown cDNAs that represent previously unrecognised TPA-regulated genes in murine skin with potential function in tumour promotion. Additionally, some TPA-induced genes, such as Sprr1A, Saa3, JunB, Il4ralpha, Gp38, RalGDS and Slpi exhibit high basal level in advanced stages of skin carcinogenesis, suggesting that at least a subgroup of the identified TPA regulated genes may contribute to tumour progression and metastasis. PMID- 12640677 TI - A novel platinum compound inhibits telomerase activity in vitro and reduces telomere length in a human hepatoma cell line. AB - Telomerase activity is detectable in most human tumors but not in most normal somatic cells or tissues. Telomerase inhibition has, therefore, been proposed as a novel and potentially selective strategy for antitumor therapy. In the present study, we found that platinum compounds, including cisplatin [cis diamminedichloro-platinum (II)], strongly inhibited the activity of partially purified rat telomerase. Among the agents tested, 2,3-dibromosuccinato [2 (methylaminomethyl)pyridine]platinum (II) (compound E) exhibited the strongest inhibition, with an median inhibitory concentration (IC(50)) of 0.8 micro M. The mode of inhibition was noncompetitive with either dNTPs or TS (first) primer, with K(i) values estimated to be 2.3 or 3.9 micro M for varied TS primer or dNTPs, respectively. Notably, cisplatin also inhibited the telomerase activity, with an IC(50) of 2.0 micro M. Again, the mode of inhibition was noncompetitive, with K(i) values estimated as 7.3 or 8.1 micro M. Preincubation of TS primer with compound E did not affect the telomerase inhibition, whereas preincubation with cisplatin caused remarkable enhancement. Treatment of a human hepatoma cell line HepG2 with a low concentration of compound E gradually reduced the telomere length, indicating that this compound was able to inhibit telomerase in living cells as well as in vitro. PMID- 12640678 TI - Suppression of EGFRvIII-mediated proliferation and tumorigenesis of breast cancer cells by ribozyme. AB - EGFRvIII is a tumor specific, ligand-independent, constitutively active variant of the epidermal growth factor receptor. Its expression has been detected in many human malignancies including breast cancer. No detectable level of EGFRvIII has, however, been observed in adult tissues, including normal breast tissues. These unique features of the EGFRvIII make it an excellent target for biologically based therapies. We have designed and generated a tumor specific ribozyme targeted to EGFRvIII. This specific EGFRvIII ribozyme is able to effectively cleave EGFRvIII mRNA under physiological conditions in a cell-free system, but does not cleave wild-type EGFR and other EGF-family receptors. While expressing this EGFRvIII-ribozyme in breast cancer cells, EGFRvIII-ribozyme is capable of downregulating endogenous EGFRvIII expression at the mRNA and protein levels. Inhibition of proliferation was observed in EGFRvIII-ribozyme transfectants. In addition, downregulation of EGFRvIII in breast cancer cells significantly inhibited tumor growth in athymic nude mice. Furthermore, this ribozyme has no effect on EGF-family receptor expression or the proliferation of breast cancer cells, which do not express EGFRvIII but express wild-type EGFR and other EGF family receptors. These results suggest that we have generated a tumor-specific, biologically functional ribozyme and further demonstrate that EGFRvIII plays a significant role in breast cancer cell proliferation. The ultimate goal of this approach is to provide a potential treatment for breast cancer by specifically targeting this receptor. PMID- 12640679 TI - Differential effects between amphoterin and advanced glycation end products on colon cancer cells. AB - Amphoterin is 1 ligand of the receptor for advanced glycation end products (RAGE). We studied expression of amphoterin and RAGE mRNA and proteins in colorectal carcinoma cells and investigated their associations with the invasive activities of cells exposed to advanced glycation end products (AGE). Expression of RAGE and amphoterin was examined in 4 colorectal carcinoma cell lines. All cell lines expressed both RAGE and amphoterin. The effects of RAGE and amphoterin on cell growth (MTT assay), migration (wound healing assay) and invasion (in vitro invasion assay) were tested by treatment of cells with RAGE and amphoterin antisense S-oligodeoxynucleotides (ODNs). Cell growth, migration and invasion were inhibited significantly in Colo320 and WiDr carcinoma cells treated with RAGE and amphoterin antisense S-ODNs compared with sense-treated cells. Differences in ligand activity between amphoterin and AGE were examined with AGE bovine serum albumin (BSA). AGE-BSA decreased cell growth, migration and invasion of amphoterin antisense S-ODN-treated Colo320 and WiDr cells compared with those of cells treated with Colo320 conditioned medium. Phosphorylation of extracellular signal-regulated kinase-1/2, Rac1 and AKT and production of matrix metalloproteinase 9 were increased to a greater degree by amphoterin than by AGE BSA. In contrast, production of inducible nitric oxide synthase and nuclear factor-kappaBp65 were increased to a greater degree by AGE-BSA than by amphoterin. PMID- 12640680 TI - Immunohistochemical analysis of p16INK4a, p14ARF, p18INK4c, p21CIP1, p27KIP1 and p73 expression in 271 meningiomas correlation with tumor grade and clinical outcome. AB - Routine pathological examination cannot distinctively predict the clinical course of meningiomas because even histologically benign tumors may recur after gross total resection. Numerous efforts have been made for the evaluation of different immunohistochemical assays in meningioma prognosis. We investigated the prognostic significance of p16INK4a, p14ARF, p18INK4c, p21CIP1, p27KIP1 and p73 expression by immunohistochemical analysis of 271 meningiomas. All tumors were additionally stained for the proliferation markers Ki-67 and DNA topoisomerase II alpha (TopoIIalpha). Significant differences between the number of p16INK4a-, p18INK4c- and p21CIP1-positive cases were noted among the 3 grades of meningiomas. p16INK4a- and p21CIP-positive tumors were found to prevail among benign meningiomas, whereas p18INK4c immunostaining was closely associated to anaplastic meningiomas. The number of p16INK4a- and p21CIP-positive cases was significantly lower in the cohort of recurrent meningiomas. In contrast, p18INK4c positive cases were clustered among recurrent meningiomas regardless of tumor grade. Immunoreactivity of p14ARF, p27KIP1 and p73 did not show any differences between meningiomas of various histology and clinical outcomes. Multivariate analysis revealed that only tumor grade and TopoIIalpha index are independent criteria for predicting meningioma recurrence. Thus, the immunohistochemical assessment of p16INK4a, p14ARF, p18INK4c, p21CIP1, p27KIP1 and p73 expression in meningiomas does not appear to provide prognostically useful information. Further studies are needed to identify more reliable prognostic markers and to address in more detail the role of cell cycle aberrations in these tumors. PMID- 12640681 TI - Hypermethylation, but not LOH, is associated with the low expression of MT1G and CRABP1 in papillary thyroid carcinoma. AB - We previously obtained gene expression profiles of 8 matched papillary thyroid carcinoma (PTC) and normal tissues using DNA microarrays. To identify novel tumor suppressor genes involved in thyroid carcinogenesis, we here analyze genes showing lower expression in PTC tumors than in normal thyroid tissues. A search for loss of heterozygosity (LOH) in 49 regions that harbor consistently down regulated genes revealed LOH in only 4 regions and in just a very small number of tumors. To determine whether the underexpression might be due to promoter methylation, we used combined bisulfite restriction analysis and bisulfite sequencing to study 7 underexpressed genes. Loss of expression of MT1G and CRABP1 is accompanied by hypermethylation in the 5' regions of these genes, but methylation was not seen in other genes tested. Combined treatment with the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (5-Aza-dC) and the histone deacetylase inhibitor trichostatin A (TSA) resulted in demethylation and re expression of the MT1G gene in the cell line K2. Treatment with 5-Aza-dC alone restored CRABP1 expression in a colorectal cancer cell line, SW48. In conclusion, LOH is a remarkably rare mechanism of loss of gene function in PTC. In contrast, hypermethylation of promoter CpG islands seems to occur at higher frequency. MT1G and CRABP1 are novel genes that are likely involved in the pathogenesis of sporadic PTC. PMID- 12640683 TI - Refined mapping of 1q32 amplicons in malignant gliomas confirms MDM4 as the main amplification target. AB - We previously reported on the amplification and overexpression of the mouse double minute 4 homolog gene (MDM4) from 1q32 in a subset of malignant gliomas (Riemenschneider et al., Cancer Res 1999;59:6091-6). More recently, amplification and overexpression of the neighboring contactin 2 gene (CNTN2) was reported in individual malignant gliomas without MDM4 amplification (Rickman et al., Cancer Res 2001;61:2162-8). To address the question of whether 1q32 carries 2 independent amplification targets or a common target other than MDM4 and CNTN2, we analyzed primary malignant gliomas for amplification and overexpression of 17 different genes from this region. Our results indicate a single region of amplification that comprises the genes MDM4, GAC1, PIK3C2B and PEPP3, with only MDM4 amplification being invariably associated with overexpression. CNTN2 was found to be coamplified with MDM4 in 3 malignant gliomas but overexpressed in only 1 of these tumors. No CNTN2 amplification was detected in any of 102 malignant gliomas without MDM4 amplification. Our data therefore corroborate the notion that MDM4 is the main amplification target on 1q32 in malignant gliomas. However, coamplification and overexpression of adjacent genes may provide an additional growth advantage in some malignant gliomas with MDM4 amplification. PMID- 12640682 TI - Analysis of chromosomal instability in pulmonary or liver metastases and matched primary hepatocellular carcinoma after orthotopic liver transplantation. AB - To investigate the genetic mechanism of metastatic spread in hepatocellular carcinoma (HCC), we analyzed genomic changes in lung or liver metastases and the corresponding primary tumors (83 tumor samples) in 18 patients who underwent orthotopic liver transplantation. We studied the incidence of microsatellite instability (MSI) and loss of heterozygosity (LOH) involving 8 highly polymorphic microsatellite markers and the polyA tract, Bat26. We also sought alterations of p53 and beta-catenin gene mutations. High MSI (>30-40% of the loci analyzed) was found only in primary tumors (11%), whereas LOH was observed in 50% of primary and in 39% of recurrent tumors. p53 mutations were found in 2 cases of primary HCC but not in the corresponding metastases. P53 was overexpressed in 4 primary HCC (22%) and 7 metastases (39%). The percentage of beta-catenin gene mutations was low (6%). Lung metastases retained the D16S402 microsatellite abnormalities observed in the primary tumors, whereas recurrent liver tumor did not (p = 0.02). In conclusion, LOH and P53 protein overexpression, rather than mutations in the p53 or beta-catenin genes or MSI, seem to be involved in the spreading of HCC, suggesting the presence of metastasis suppressor genes in the vicinity of the chromosomal loci in question. PMID- 12640684 TI - Association between self-reported environmental tobacco smoke exposure and lung cancer: modification by GSTP1 polymorphism. AB - Environmental Tobacco Smoke (ETS) exposure has been associated with lung cancer risk. ETS is composed of emissions from cigarette smoke and contains a higher concentration of tobacco smoke carcinogens than mainstream smoke. Polymorphisms in genes that metabolize tobacco smoke carcinogens have been studied as effect modifiers of the association between active smoking and lung cancer risk. GSTP1 is a polymorphic gene that encodes for GST pi, a detoxification enzyme and has a high expression in the lung. We investigated the association between ETS and lung cancer risk and the modification of this association by the GSTP1 polymorphism. Using a case-control design, individuals were genotyped for GSTP1 using PCR-RFLP techniques. All analyses were carried out using multiple logistic regression. The association between ETS exposure and lung cancer risk was evaluated in different strata based on smoking habits to evaluate the consistency of results. The effect of the GSTP1 polymorphisms on lung cancer risk was evaluated by considering the joint effect of having both an ETS exposure and the GSTP1 GG genotype compared to the absence of ETS exposure and the GSTP1 AA genotype as a reference group as well as doing stratified analysis by genotype. ETS exposure was associated consistently with higher lung cancer risk in all the strata considered. The adjusted odds ratios (AOR) evaluating the association between ETS and lung cancer risk for the different strata were: nonsmokers (Cases/Controls 66/413; AOR = 1.38; 95% CI = 0.78-2.43), ex-smokers (Cases/Controls 560/527; AOR = 1.66; 95% CI = 1.22-2.25), current smokers (Cases/Controls 415/219; AOR = 1.56; 95% CI = 1.00 2.41). The AORs for ex-smokers and light smoking subgroups were: ex-smokers who quit for 19 years or more (Cases/Controls 144/244; AOR = 2.64; 95% CI = 1.55 4.50), ex-smokers who quit for 10-19 years (Cases/Controls 141/128; AOR = 1.16; 95% CI = 0.66-2.04), ex-smokers who quit for 10 years or less (Cases/Controls 247/122; AOR = 1.45; 95% CI = 0.83-2.55) and participants who had <15 packyears and nonsmokers combined (Cases/Controls 143/640; AOR = 1.52; 95% CI = 1.02-2.28). Among those with the GSTP1 GG genotype the ETS-lung cancer risk association was greater than those with the GSTP1 AA genotype: nonsmokers (GSTP1 GG AOR = 7.84; 95% CI = 0.80-76.68; GSTP1 AA AOR = 1.15; 95% CI = 0.46-2.90), ex-smokers (GSTP1 GG AOR = 2.32; 95% CI = 0.90-5.96; GSTP1 AA AOR = 2.15; 95% CI = 1.34-3.44), current smokers (GSTP1 GG AOR = 1.75; 95% CI = 0.42-7.32; GSTP1 AA AOR = 1.32; 95% CI = 0.67-2.58) and participants who had <15 packyears and nonsmokers (GSTP1 GG AOR = 1.93; 95% CI = 0.54-6.97; GSTP1 AA AOR = 1.58; 95% CI = 0.83-3.01). We found that ETS exposure is associated with higher lung cancer risk. Furthermore, the presence of the GSTP1 GG genotype appears to enhance the magnitude of the association between ETS exposure and lung cancer. Larger studies will be needed to confirm these preliminary findings. PMID- 12640685 TI - Incidence trends and familial risks in invasive and in situ cutaneous melanoma by sun-exposed body sites. AB - We studied incidence trends, age-incidence relationships and familial risks in invasive and in situ cutaneous melanoma, based on the Swedish Family-Cancer Database of more than 10 million individuals. Offspring were 0-66 years of age. Cancers were obtained from the Swedish Cancer Registry from years 1961-98. The study was based on 9,771 offspring and 22,888 parents with invasive melanoma and 2,446 offspring and 5,017 parents with in situ melanoma. Incidence rates increased markedly for invasive melanoma in the trunk. For in situ melanoma, trunk and head and neck were affected, and, in addition, legs for women. The maximal incidence was around age 80 years, independent of the type or site in men; in women early onset superficially spreading melanoma shifted the age for maximal incidence to about 60 years. For in situ melanoma, lentigo maligna was the main histogenetic type in the head and neck but in the trunk and legs superficially spreading melanoma was somewhat more common. Standardized incidence ratios (SIR) were calculated for familial risk at exposed and covered sites. The combined familial risks for invasive and in situ melanoma were higher at covered (SIR 3.56 from parents) than sun-exposed (1.92 from parents) sites and they agreed when familiality was defined between parents and offspring or between siblings; the sibling SIRs were 3.90 at covered and 2.53 at exposed sites. The data suggest that the higher melanoma density at exposed sites masks familial effects. Furthermore, sun exposure does not appear to reinforce the familial effect. PMID- 12640686 TI - Childhood cancer in relation to infections in the community during pregnancy and around the time of birth. AB - In a retrospective cohort study of 404,106 live births in the northern region of England, 1975-1986, we investigated whether higher levels of community infections during the mother's pregnancy and in early life were risk factors for cancer, by diagnostic group (leukaemia and non-Hodgkin's lymphoma, Hodgkin's disease, brain/spinal tumours, neuroblastoma, other tumours), diagnosed 1975-2001 under age 15 years. Logistic regression was used to relate risk to measures of community infections (measles, respiratory and other infections) in 3 prenatal and 2 postnatal quarters. There was an increased risk of Hodgkin's disease among children exposed around birth to higher levels of measles (odds ratio for trend = 2.3, 95% confidence interval 1.3-4.2, p = 0.01). For other diagnostic groups, there was no consistent evidence of an association between risk and exposure to infections. Although the significant association observed for Hodgkin's disease may be a chance finding, consequent to multiple hypothesis testing or the ecologic nature of the study, it is consistent with other recent epidemiologic results suggesting that the risk of Hodgkin's disease may be associated with exposure to infections. PMID- 12640687 TI - Important increase of invasive lobular breast cancer incidence in Geneva, Switzerland. AB - A recent paper from the United States reported a sharp and unexplained increase in invasive lobular breast cancer incidence since 1977 (Li et al., Cancer 2000;88:2561-9). We investigated if this trend was also present in Geneva, Switzerland, where breast cancer incidence is one of the highest in Europe. We analyzed trends in breast cancer incidence according to histologic subtype, age and stage, to clarify the pattern. Our population-based study includes all histologically confirmed invasive breast carcinomas (n = 6,247) recorded between 1976 and 1999 at the Geneva Cancer Registry. Breast histology was classified as ductal carcinoma, lobular carcinoma and other. Incidence trends were studied by log-linear regression analyses. Models including effects of age, period and birth cohorts were used to describe rising incidence trends. The incidence of ductal carcinoma increased 1.2% per year (p(trend) < 0.001) from 85.2 to 110.1/100,000. This increase concerned women aged 50-69 years and early-stage tumors. Lobular cancer incidence increased disproportionately (14.4% per year, p(trend) < 0.01) and rose from 2.9 to 20.5/100,000. This increase affected all age categories and both localized and advanced stages. In addition, a strong age-cohort effect was present (p < 0.05), and women aged 50-59 years born after 1944 experienced the most marked increase. Our study shows a disproportionate increase of lobular breast cancer incidence compared to ductal cancer incidence. Contrary to ductal cancer, trends for lobular cancer are unlikely to be explained by increased use of screening mammography. Other explanations must be researched, in particular the role played by hormone replacement therapy. PMID- 12640688 TI - Photodynamic therapy with Pd-Bacteriopheophorbide (TOOKAD): successful in vivo treatment of human prostatic small cell carcinoma xenografts. AB - Small cell carcinoma of the prostate (SCCP), although relatively rare, is the most aggressive variant of prostate cancer, currently with no successful treatment. It was therefore tempting to evaluate the response of this violent malignancy and its bone lesions to Pd-Bacteriopheophorbide (TOOKAD)-based photodynamic therapy (PDT), already proven by us to efficiently eradicate other aggressive non-epithelial solid tumors. TOOKAD is a novel bacteriochlorophyll derived, second-generation photosensitizer recently, developed by us for the treatment of bulky tumors. This photosensitizer is endowed with strong light absorbance (epsilon(0) approximately 10(5) mol(-1) cm(-1)) in the near infrared region (lambda=763nm), allowing deep tissue penetration. The TOOKAD-PDT protocol targets the tumor vasculature leading to inflammation, hypoxia, necrosis and tumor eradication. The sensitizer clears rapidly from the circulation within a few hours and does not accumulate in tissues, which is compatible with the treatment of localized tumor and isolated metastases. Briefly, male CD1-nude mice were grafted with the human SCCP (WISH-PC2) in 3 relevant anatomic locations: subcutaneous (representing tumor mass), intraosseous (representing bone metastases) and orthotopically within the murine prostate microenvironment. The PDT protocol consisted of i.v. administration of TOOKAD (4 mg/kg), followed by immediate illumination (650-800 nm) from a xenon light source or a diode laser emitting at 770 nm. Controls included untreated animals or animals treated with light or TOOKAD alone. Tumor volume, human plasma chromogranin A levels, animal well being and survival were used as end points. In addition, histopathology and immunohistochemistry were used to define the tumor response. Subcutaneous tumors exhibited complete healing within 28-40 days, reaching an overall long-term cure rate of 69%, followed for 90 days after PDT. Intratibial WISH-PC2 lesions responded with complete tumor elimination in 50% of the treated mice at 70-90 days after PDT as documented histologically. The response of the orthotopic model was also analyzed histologically with similar results. The study with this model suggests that TOOKAD-based PDT can reach large tumors and is a feasible, efficient and well-tolerated approach for minimally invasive treatment of local and disseminated SCCP. PMID- 12640690 TI - Hodgkin's disease etiology and novel viruses: clues from groups exposed to blood products. PMID- 12640689 TI - New prognostic indicator for non-small-cell lung cancer, quantitation of thymidylate synthase by real-time reverse transcription polymerase chain reaction. AB - Thymidylate synthase (TS) is an enzyme that catalyzes an important DNA biosynthesis process. The gene expression of TS has not been reported in non small-cell lung cancer (NSCLC) patients. To clarify the correlation between TS mRNA levels and clinicopathological features of NSCLC, we examined 70 Stage I and II NSCLC patients for intra-tumoral expression of TS using TaqMan reverse transcription polymerase chain reaction (RT-PCR) assay and immunohistochemistry methods. We also investigated the TS promoter 28 bp polymorphism in 48 cancer tissues using PCR amplification of genomic DNA. Lung cancer tissue showed higher TS mRNA levels than normal lung tissue (Mann-Whitney U-tests; p = 0.0020). Further, TS mRNA expression was correlated with immunohistochemical TS expression (p = 0.029). We obtained 2 different DNA fragments, which indicated triple-repeat (3R) and double-repeat (2R) type alleles. Cancer tissues with the 3R/3R genotype showed significantly higher TS mRNA levels as compared to those with other genotypes (p = 0.0019). The TS genotype was also correlated with immunohistochemical TS expression (chi(2) test; p = 0.0079). The disease-free survival of the low TS mRNA level group was significantly better than those with high TS mRNA levels (log-rank test; p = 0.010), however, there were no significant differences found by immunohistochemical evaluation (p = 0.34) or TS genotype analysis (p = 0.11). A multivariate analysis revealed that high TS mRNA levels independently contributed to disease-free survival. The quantitation of TS mRNA levels is clinically more sensitive and useful for determining the prognosis of Stage I and II NSCLC patients than an immunohistochemical evaluation. PMID- 12640699 TI - [Research on genetic engineering vaccine of Helicobacter pylori]. PMID- 12640700 TI - [Construction of prokaryotic expression system of ureB gene from a clinical isolate of Helicobacter pylori and identification of immunogenicity of the fusion protein]. AB - OBJECTIVE: To clone Helicobacter pylori ureB gene, to construct prokaryotic expression system of the gene and to identify immunogenicity of the fusion protein. METHODS: The ureB gene from a clinical isolate Y06 of H.pylori was amplified by high fidelity PCR. The nucleotide sequence of the target DNA amplification fragment was sequenced after T-A cloning. The expression vector pET32a with inserted ureB gene was constructed. ureB fusion protein was expressed in E.coli strain BL21DE3 induced by IPTG at different dosages. Western blot using antibody against whole cell of H.pylori as well as immunodiffusion assay using antiserum of rabbit against the fusion protein was applied to determine immunogenicity of the fusion protein. RESULTS: In comparison with the reported corresponding sequences, the homology of nucleotide sequence of the cloned ureB gene was from 96.88% approximate, equals 97.82%, while the homology of its putative amino acid sequence was as high as 99.65% approximate, equals 99.82%. The expression output of UreB protein in pET32a-ureB-BL21DE3 system was approximately 40%of the total bacterial proteins. UreB protein was able to combine with antibody against whole cell of H.pylori and induce rabbit to produce high titer antibody after the animal was immunized with the protein. CONCLUSION: An expression system with high efficiency of H.pylori ureB gene has been established successfully. The expressed UreB protein with satisfactory immunogenicity and immunoreactivity can be used as antigen in H.pylori vaccine. PMID- 12640701 TI - [Cloning, expression and identification of hpaA gene from a clinical isolate of Helicobacter pylori]. AB - OBJECTIVE: To clone Helicobacter pylori adhesin (hpaA) gene,to construct the expression vector of the gene and to identify immunogenicity of the fusion protein. METHODS: The hpaA gene from a clinical isolate Y06 of H.pylori was amplified by high fidelity PCR. The nucleotide sequence of the target DNA amplification fragment was sequenced after T-A cloning. The expression vector pET32a with inserted hpaA gene was constructed. hpaA fusion protein was expressed in E.coli strain BL21DE3 induced by IPTG at different dosages. Western blot using antibody against whole cell of H.pylori as well as immunodiffusion assay using antiserum of rabbit against the fusion protein was applied to determine immunogenicity of the fusion protein. RESULTS: In comparison with the reported corresponding sequences, the homology of nucleotide sequence of the cloned hpaA gene was from 94.25% approximate, equals 97.32%, while the homology of its putative amino acid sequence was as high as 95.38% approximate, equals 98.46%. The expression output of HpaA fusion protein in pET32a-hpaA-BL21DE3 system was approximately 40% of the total bacterial proteins. HpaA fusion protein was able to combine with antibody against whole cell of H.pylori and induce rabbit to preduce high titer antibody after the animal was immunized with the protein. CONCLUSION: An expression system with high efficiency of H.pylori hpaA gene has been established successfully. The expressed HpaA fusion protein with satisfactory immunogenicity and immunoreactivity can be used as antigen in H.pylori vaccine. PMID- 12640702 TI - [Cloning, expression and identification of flaB gene from a clinical isolate of Helicobacter pylori]. AB - OBJECTIVE: To clone Helicobacter pylori flagellin B gene (flaB) to construct prokaryotic expression system of the gene and to identify immunity of the fusion protein. METHODS: The flaB gene from a clinical isolate Y06 of H.pylori was amplified by high fidelity PCR. The nucleotide sequence of the target DNA amplification fragment was sequenced after T-A cloning. The expression vector pET32a with inserted flaB gene was constructed. FlaB fusion protein was expressed in E.coli strain BL21DE3 inducted by IPTG at different dosages. Western blot was applied to determine immunoreactivity and immunogenicity of the fusion protein and antibody against whole cell of H.pylori and rabbit antiserum immunized with the fusion protein, respectively. RESULTS: In comparison with the reported corresponding sequences, the homology of nucleotide sequence of the cloned flaB gene was from 96.31% approximate, equals 97.73%, while the homology of its putative amino acid sequence was as high as 99.41% approximate, equals 100%. The expression output of FlaB fusion protein in pET32a-flaB-BL21DE3 system was approximately 40% of the total bacterial proteins. FlaB fusion protein was able to combine with antibody against whole cell of H.pylori and induce rabbit to produce specific antibody with high titer after the animal was immunized with the protein. CONCLUSION: A prokaryotic expression system of H. pylori flaB gene with high efficiency has been established successfully. The expressed FlaB fusion protein with satisfactory immunogenicity and immunoreactivity can be used as antigen in H.pylori vaccine and detect kit. PMID- 12640704 TI - [Establishment of Helicobacter pylori infection model in Mongolian gerbil]. AB - OBJECTIVE: To establish a stable and reliable model of Helicobacter pylori infection in Mongolian gerbil and to observe pathological changes in gastric mucosa from the infected animals. METHODS: Mongolian gerbils were randomly divided into six groups infected with H.pylori strain NCTC11637 (n=6, group N), six groups infected with H.pylori clinical strain Y06 (n=6, group Y) and six groups as negative control (n=4, group C). H.pylori suspensions at the concentrations of 2 X 10(8)CFU/ml and 2 X 10(9) CFU/ml of strain NCTC11637 and strain Y06 were prepared with Brucella broth from Columbia agar containing sheet blood. The animals in one group N and in one group Y were orally challenged once with 0.5 ml of 2 X 10(8) CFU/ml H.pylori suspension. The animals in another group N and in another group Y were orally challenged with 0.5 ml of 2 X 10(9) CFU/ml H.pylori suspension for three times at the intervals of 24 hours, respectively. The animals were killed after 2nd, 4th and 6th week of the last infection and the gastric mucosal samples were taken for urease test, bacterial isolation, routine pathological and H.pylori histochemical examinations. RESULTS: Infection rates of the animals in group N and group Y at the 2nd, 4th and 6th week after one challenge were 0%, 0%, 66.7% and 0%, 16.7%, 16.7%, respectively. Infection rates of the animals in groups N and Y at the 2nd, 4th and 6th week after three challenges were 66.7%, 100%, 100% and 66.7%, 66.7%, 100%, respectively. In animals with positive bacterial isolation H.pylori was found to colonized on the surface of gastric mucosal cells and in the gastric pits, and the lamina propria of gastric mucosal was infiltrated with chronic inflammatory cells. CONCLUSION: By using H.pylori suspension at high concentration of 1 X 10(9) CFU for multiple times, the orally challenged Mongolian gerbils can be prepared as a stable and reliable H.pylori infection model. H.pylori can colonize in gastric mucosa of the infected animals, and mild inflammation reactions can be seen. PMID- 12640703 TI - [Cloning, expression and identification of Escherichia coli LTB gene and Vibrio cholerae CTB gene]. AB - OBJECTIVE: To clone the LTB gene of E.coli and the CTB gene of V.cholerae, and to construct expression vectors of these genes. METHODS: The LTB gene from E.coli strain 44815 and the CTB gene from V.cholerae strain eastern 74 were amplified by high fidelity PCR. The nucleotide sequences of the two target DNA amplification fragments were sequenced after T-A cloning. pET32a expression vectors with inserted LTB and CTB genes were constructed. The LTB and CTB fusion proteins were expressed in E.coli strain BL21DE3 inducted by IPTG at different dosages. The two expression products were identified by SDS-PAGE and G(M1)-ELISA. RESULTS: In comparison with the reported LTB and CTB sequences, the nucleotide sequence homologies of the cloned LTB gene and CTB gene were from 99.12% approximate, equals 99.71% and 98.54% approximate, equals 99.42%, while their putative amino acid sequence homologies were as high as 97.58% approximate, equals 99.19% and 96.77% approximate, equals 99.19%. The expression outputs of LTB and CTB fusion proteins in pET32a LTB BL21DE3 and pET32a-CTB-BL21DE3 systems were approximately 30% and 10% of the total bacterial proteins, respectively. The LTB and CTB fusion proteins were able to combine with bovine G(M1) confirmed by ELISA. CONCLUSION: The expression systems of LTB and CTB genes have been successfully established. Both the expressed LTB and CTB fusion proteins possess mucosal adjuvant immunoactivity. PMID- 12640705 TI - [Analysis on the vacA dominant genotypes and their nucleotide sequences of Helicobacter pylori isolates in Zhejiang area]. AB - OBJECTIVE: To determine vacA dominant genotypes of Helicobacter pylori in patients with peptic ulcer (PU) or chronic gastritis (CG). METHODS: H.pylori strains were isolated from mucosa samples of gastric antrum and corpus of patients suffering from PU (n=29) or CG (n=34), 126 strains of H.pylori were selected for PCR to detect s and m regions in vacA gene of the isolates. Parts of the amplification products were sequenced after T A cloning. The correlation between infection or coinfection with different vacA genotypes of H.pylori and different gastroduodenal diseases was further analyzed. RESULTS: The positive amplification products of vacA gene, s and m regions, were found in the DNA samples of all the isolates. In these products, s1a/m1, s1a/m2, s1a/m1b and s1a/m1b m2 genotypes of vacA gene were detected and s1b and s2m1a genotypes absent. Proportions of the s1a/m1, s1a/m2, s1a/m1b and s1a/m1b-m2 genotypes were 7.1% (9/126), 61.9% (78/126), 29.4% (37/126) and 1.6% (2/126), respectively. 17.5% (11/63) of the patients were confirmed to be coinfected with different genotype H.pylori strains. No statistical differences were found in the distribution of different genotype H.pylori strain infection in the gastric diseases (P>0.05). In comparison with the reported sequences of H.pylori strain 60190 with s1a genotype and strain 87-203 with m2 genotype, homologies of the nucleotide sequences of s1a PCR products from 6 strains of H.pylori isolates and m2 PCR products from 4 strains of H.pylori isolates were 93.15% approximate, equals 94.86%and 93.63% approximate, equals 97.61%, respectively. CONCLUSION: H.pylori with s1a/m2 or s1a/m1b are the dominant genotypes in the PU or GC patients in Zhejiang area. The nucleotide sequences of partial amplification products from the vacA dominant genotypes of H.pylori show high homology compared with the reported sequences. Part of the patients may be coinfected with different vacA genotypes of H.pylori. PMID- 12640706 TI - [Analysis of babA2 cagA and vacA genotypes of Helicobacter pylori in chronic gastritis and peptic ulcer]. AB - OBJECTIVE: To investigate the distribution of babA2 cagA and vacA genotypes of Helicobacter pylori (Hp) in chronic gastritis and peptic ulcer and to discuss the relationship between babA2, cagA and vacA genotypes of Hp and chronic gastritis and peptic ulcer. METHODS: babA2, cagA genotypes and vacA subtype of 58 Hp strains isolated from patients of Zhejiang province with chronic gastritis or peptic ulcer were tested by polymerase chain reaction. RESULTS: The positive rates of babA2, cagA, vacAs1a, vacA m1 and vacA m2 of 58 Hp strains were 87.9%, 100%, 93.1%, 1.7% and 65.5%, respectively. There were no significant differences between the positive rates of babA2, vacA s1a and vacA m2 genes of Hp strains isolated from patients with chronic gastritis and peptic ulcer. CONCLUSION: The genotypes of Hp isolated from patients of Zhejiang province were predominatly babA2 positive, cagA positive and vacA s1a/m2. The relationships between babA2, cagA and vacA genotypes of Hp and chronic gastritis and peptic ulcer can not be identified. PMID- 12640707 TI - [Preparation of monoclonal antibodies against oh(8)dG and their biological characteristics]. AB - OBJECTIVE: To prepare monoclonal antibodies against oh(8)dG and to evaluate the relationship between Hp infection and oxidative DNA damage by detecting oh8dG in gastric mucosa. METHODS: BALB/C mice were immunized with BSA-oh(8)dG conjugate, monoclonal antibodies were prepared by hybridoma technique, the biological characteristics of antibodies were analysed by competitive ELISA, Western blot and immunohistochemistry. RESULTS: Two strains of hybridoma cell were obtained. ELISA and Western blot indicated that the antibodies were fairly specific for oh(8)dG. In immunohistochemistry,the positive rate of oh(8)dG expression in Hp positive tissues and Hp negative tissues was 55% and 5%, respectively(P<0.01). CONCLUSION: The prepared antibodies can specially recognize oh(8)dG and immunohistochemistry with the monoclonal antibodies showed Hp infection can increase oh(8)dG level in gastric mucosa. PMID- 12640708 TI - [Correlation of rdxA gene mutation and metronidazole resistance of Helicobacter pylori]. AB - OBJECTIVE: To demonstrate the correlation of rdxA gene mutation and metronidazole (MTZ) resistance of H.pylori isolates in the local area. METHODS: Clinical strains of H.pylori were isolated from gastric biopsy of patients. Resistance to metronidazole of the isolates was determined by using diffusion test and two fold dilution test. Genome DNAs of the isolates were prepared for PCR to detect rdxA gene. The target amplification products were sequenced after T-A cloning. The sequences were compared with the reported sequences from Hp26695 and 134 other strains of H.pylori. RESULTS: MTZ resistance rate was 76.1% in 21 clinical isolates. The target fragment 886 bp in length containing rdxA gene could be successfully amplified. In comparison with the reported corresponding sequence of H.pylori stain 26695, homologies of the nucleotide sequences from the amplification products were 90.1% approximate, equals 95.1%. Mutations caused by base insertion/deletion and substitution in the MTZ resistance isolates were found. Among these mutations, two types of insertion mutations have not been reported in literatures. No same mutations were present in the MTZ sensitive isolates. CONCLUSION: The rdxA gene mutation may play an important role in MTZ resistance of H.pylori. PMID- 12640709 TI - [Lipid-peroxidantion damage of embryo and placenta induced by artesunate in rats]. AB - OBJECTIVE: To examine the effect and mechanism of artesunate (Art) on embryo development. METHODS: Rat embryo and placental glutathione peroxidase (GSH-Px)and malondialdehyde (MDA) were identified by using DTNB (dithionitrobenzene) direct method and TBA (thiobarbituric acid). We investigated the damage of decidual cells caused by Art using cell culture techniques. RESULTS: Subcutaneous administration of Art in rats on d 6 approximate, equals d 10 of gestation induced developmental toxicity. Absorption increased when progressively increased doses were given (r=0.996,P<0.01). Twenty four hours post injection, GSH-Px in embryo decreased significantly while MDA content was significantly higher than that in the control group (P<0.05). GSH Px: study group was(43.7+/ 10.7)micromol/min.mg(-1)Hb, control group was(54.5+/-10.1)micromol/min.mg(-1)Hb; MDA:study group was(230.2+/-19.8)nmol/g tissue, control group was(150.4+/ 44.1)nmol/g tissue. Placental GSH-Px activity was significantly higher than that in the control group(P<0.01). After cultured human decidual cells were exposed to Art for 24 h, the LC50 was (25.2+/-3.5)mg/L. CONCLUSION: Art may induce developmental toxicity in rat embryo and placenta by neutralizing the antioxidant defense mechanism. PMID- 12640710 TI - [Population pharmacokinetics of phenytoin in pediatric patients]. AB - OBJECTIVE: To study population pharmacokinetics of phenytoin in pediatric patients by using sparse data. METHODS: We used routinely collected therapeutic drug monitoring data, derived from the steady state serum concentrations of phenytoin in 42 pediatric outpatients with epilepsy. Depending on whether the patients were administered with phenytoin alone or coadministered with phenobabital or clonazepam, the subjects were divided into two groups: phenytoin group and coadministration group. The population parameter and individual parameter of phenytoin in children were estimated using Monte Carlo method. RESULTS: The children's phenytoin population pharmacokinetic parameters Vm and Km were 9.8 mg.kg(-1).d(-1) and 2.73 mg/L in phenytoin group; and 9.2 mg.kg(-1).d( 1) and 3.24 mg/L in coadministration group. There were good relationship between predicted and determined concentrations with correlation coefficient of 0.999 and 0.984, respectively. CONCLUSION: The coadministration of phenobarbital or clonazepam obviously affected the pharmacokinetics of phenytoin. The population pharmacokinetics of phenytoin in children may provide a usefull index for individualization of dosage regimen. PMID- 12640711 TI - [Improvement of cardiomyocyte isolation for different purposes]. AB - OBJECTIVE: To improve the methods of isolating single ventricular myocytes for purposes of different experiments. METHODS: The cardiac ventricular myocytes were isolated enzymatically by Langendorff perfusion technique at constant flow rate. The patch clamp whole-cell recording, a spectrofluorometric method and a video tracking system were used to verify the basic electrophysiological properties, intracellular calcium transient and cell contraction of the single myocytes. RESULTS: The resting membrance potential of the myocyte was (-74.06+/-4.54)mV. The whole-cell sodium, potassium and calcium currents, intracellular calcium transients and contractile properties were stable and typical. CONCLUSION: The myocytes obtained by adjusting perfusion parameters are well suited for electrophysiological recording, intracellular calcium and single cell contraction measurements. PMID- 12640712 TI - [Serum levels of soluble intercellular adhesion molecule-1 in patients with cerebral infarct]. AB - OBJECTIVE: To observe the changes of the levels of soluble intercellular adhesion molecule-1(sICAM-1) in serum of patients with cerebral infarct and to explore the effect of sICAM-1 on cerebral infarct. METHODS: The serum levels of sICAM-1 in 55 patients with cerebral infarct both in acute stage(within 2 days) and convalescence(2 weeks after attack) were detected by using ELISA. At the same time, we compare the results with those of 32 patients having other neurologic diseases(20 patients with sciatica, 12 with trigeminal neuralgia) and 30 healthy subjects. RESULTS: (1) The serum levels of sICAM-1 of patients with cerebral infarct (acute stage: 766+/-179 microgram/L, convalescence: 602+/-155 microgram/L, respectively) were significantly higher than those of the control groups(530+/-77 microgram/L and 521+/-116 microgram/L, respectively, P<0.01). (2)There was a positive correlation of SICAM levels with the amount of leukocytes in acute stage(r=0.285,P<0.05), but negative correlation to clinical severity of cerebral infarct(r= 0.333,P<0.05). And there was no significant correlation between the level of sICAM-1 and the levels of cholesterol and triglyceride in serum(r= 0.042 and r=0.061, respectively, P>0.05). (3)There was no significant difference between sICAM levels of patients of cerebral cortex infarct and those of patients with basal ganglia infarct(773+/-178 microgram/L and 758+/-183 microgram/L, respectively, P>0.05). (4)The levels of sICAM-1 between patients of cerebral infarct with or without hypertension were no significant difference(774+/-189 microgram/L and 754+/-165 microgram/L, respectively, P>0.05). CONCLUSION: The levels of sICAM-1 increase significantly in patients with cerebral infarct. sICAM-1 may participate in the pathophysiologic process through inflammatory mechanism. PMID- 12640713 TI - [Experimental study on the relationship between insulin and hypertension]. AB - OBJECTIVE: To investigate the association between insulin and hypertension. METHODS: Twenty spontaneously hypertension rats(SHR) and twenty Wistar-Kyoto rats(WKY) were randomly divided into two groups control group and insulin injection group. Systolic blood pressure(SBP),pulse rate(P), fasting blood sugar(FBS), fasting serum insulin(FINS) and insulin sensitivity index(ISI) were observed or calculated before and at the 60th day of the experiment. RESULTS: (SBP 165.5+/-5.8 compared with 108.2+/-4.7mmHg,P<0.01), P(405.8+/-19.6 compared with 336.6+/-9.6 /min P<0.01), FINS (6.89+/-0.99 compared with 5.78+/ 0.91mU/L,P<0.05)of SHR were higher than those of WKY before experiment, but there was a lower ISI of SHR -3.397+/-0.191 compared with -3.085+/-0.132,P<0.01 . There were increases of SBP(210.0+/-8.5 compared with 184.3+/-8.0 mmHg,P<0.01),P(452.2+/-13.9 compared with 406.0+/-22.7/min P<0.01) and FINS (28.37+/-3.86 compared with 7.32+/-0.87 mU/L,P<0.01) in insulin injection group of SHR than those in controls,but ISI -4.119+/-0.260 compared with -3.604+/-0.174 P<0.01 decreased in insulin injection group; The same changes were observed in WKY rats after insulin injection (131.6+/-6.7 compared with 110.4+/-5.1 mmHg, 378.2+/-13.2 compared with 347.1+/-14.9/min 22.64+/-2.13 compared with 5.55+/ 0.77 mU/L,-3.474+/-0.214 compared with 3.094+/-0.191 P<0.01 respectively). CONCLUSION: Insulin resistance and hyperinsulinemia exist in SHR,chronic hyperinsulinemia may increase SBP and P,decrease ISI of WKY and SHR. PMID- 12640714 TI - [Microvessel density, epithelial-stromal vascular cuffing and expression of vascular endothelial growth factor in human cervical carcinoma]. AB - OBJECTIVE: To observe microvessel density(MVD), epithelial stromal vascular cuffing(VC) and expression of vascular endothelial growth factor(VEGF) in human cervical carcinomas and to clarify their significance in the invasion and metastasis of cervical carcinoma. METHODS: VEGF and CD34 were stained immunohistochemically (SP) in 57 cases of cervical carcinoma (30 cases of squamous cell carcinoma, 20 of adenocarcinoma 7 of glandular and squamous cell carcinoma), 29 cases of cervical intraepithelial neoplasia (CIN) and 16 cases of normal cervices, meanwhile, MVD and VC were also assayed. RESULTS: There were significant differences among the above 5 groups for MVD P<0.01 . The VC pattern showed a significant difference between cervical carcinoma and CIN or control group P<0.01). The positive rates of VEGF in normal cervical epithelium, CIN, squamous cell carcinoma, adenocarcinoma, glandular and squamous cell carcinoma were 18.8% 3/16, 82.8% 24/29), 93.3% 28/30), 100% 20/20 and 7/7(100%), respectively. There were significant differences between these cervical lesion groups and the control group(P<0.001). The MVD showed significant differences between the positive pelvic node metastasis and negative pelvic node metastasis P<0.05). There was no significant correlation between the expression of VEGF and the tumor diameter, clinical stage, pathologic grade and pelvic node metastasis. CONCLUSION: The expression of VEGF may play an important role in the angiogenesis of cervical carcinoma. Degree of malignancy of cervical carcinoma has a close association with microvessel density. PMID- 12640715 TI - [Detection of P-170 and treatment with berbamine in primary acute leukemia]. PMID- 12640716 TI - [Prospective study of preoperative transcatheter arterial chemo-embolization for Wilms'tumor]. PMID- 12640717 TI - [Preserving anterior lens capsule on pars plana lensectomy-vitrectomy in ocular injury]. PMID- 12640718 TI - [The CT manifestation of endometriotic cyst ovary]. PMID- 12640719 TI - [Progress in the use of transgenic plants as vaccine production systems]. PMID- 12640720 TI - [Dendritic cell and autoimmune diseases]. PMID- 12640721 TI - Tumor necrosis factor-alpha and interferon-gamma directly impair epithelial barrier function in cultured mouse cholangiocytes. AB - BACKGROUND/AIMS: In primary biliary cirrhosis (PBC), cytokines from CD4+ T lymphocytes were suggested to contribute to the intralobular bile duct damage together with cellular immunity by CD8+ T lymphocytes. Recently, we reported that immunolocalization of 7H6--a tight junction (TJ)-associated protein--was significantly diminished in cholangiocytes in the PBC liver. In this study, we examined the direct effects of several cytokines--tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), interleukin-2 and 4 (IL-2 and 4)--on TJ in immortalized mouse cholangiocytes. Moreover, we examined the inhibitory effect of ursodeoxycholic acid (UDCA) on cytokine-induced changes in paracellular permeability. METHODS: Barrier function of TJ was evaluated by measuring transepithelial electrical resistance (TER) and 3H-inulin flux. We also performed immunostaining and immunoblotting for TJ-associated proteins--claudin-1 and -3, occludin, zonula occluden-1 (ZO-1) and 7H6. RESULTS: TNF-alpha and IFN-gamma, but neither IL-2 nor IL-4, significantly decreased TER (P < 0.005). 3H-inulin flux studies confirmed IFN-alpha-induced increases in paracellular permeability of cholangiocytes (P < 0.001). In immunostaining and immunoblotting studies, TJ associated proteins were well preserved in TNF-alpha- or IFN-gamma-treated cells. Ursodeoxycholic acid has been found to have no inhibitory effect on increased paracellular permeability induced by TNF-alpha or IFN-gamma. CONCLUSION: These findings show that TNF-alpha and IFN-gamma disrupt barrier function of TJ in cholangiocytes without major structural changes to TJ and suggest that disruption of TJ function and subsequent leakage of the bile constituents may influence the aggravation of cholestasis in PBC. PMID- 12640723 TI - Effect of in situ hypothermic perfusion on intrahepatic pO2 and reactive oxygen species formation after partial hepatectomy under total hepatic vascular exclusion in pigs. AB - AIM: This study examined attenuation of ischemia and reperfusion (I/R) induced liver injury during liver resections by hypothermic perfusion of the liver under total hepatic vascular exclusion (THVE). METHOD: Reactive oxygen species (ROS) formation, microcirculatory integrity and endothelial cell damage were investigated. Left hemihepatectomy (LHX) was performed without in situ perfusion (control-LHX, n = 5) or with concomitant in situ perfusion with hypothermic (4 degrees C) Ringer-glucose (cold-LHX, n = 5) or normothermic (38 degrees C) Ringer glucose (warm-LHX, n = 5). Glutathione (GSH) and malondialdehyde (MDA) concentrations, tissue pO2 levels and hyaluronic acid (HA) uptake capacity were determined. RESULTS: After cold, warm and control-LHX, 24 h survival was 5/5, 0/5 and 3/5, respectively. GSH levels were best preserved after cold-LHX during reperfusion. MDA levels increased in all groups without significant differences between the groups during reperfusion. Tissue pO2 levels increased after cold-LHX whereas after warm-LHX and control-LHX, pO2 levels decreased during reperfusion. HA uptake capacity remained normal after cold-LHX. After warm-LHX and control LHX, HA uptake capacity decreased after 6 h of reperfusion but recovered after 24 h of reperfusion in the control-LHX group. CONCLUSION: Moderate hypothermic perfusion protects the liver from I/R injury during LHX under THVE. This protective effect depended on maintenance of liver microcirculation rather than a reduction in ROS formation. PMID- 12640722 TI - Previous hepatitis B virus infection is associated with worse disease stage and occult hepatitis B virus infection has low prevalence and pathogenicity in hepatitis C virus-positive patients. AB - BACKGROUND: Anti-hepatitis C virus (anti-HCV) patients with chronic liver disease (CLD) frequently show markers of previous hepatitis B virus (HBV) infection. Moreover, they may carry occult HBV infection. These features might influence clinical and biochemical features as well as stage of disease. AIM: To assess the prevalence and clinical associations of previous (positivity for anti-HBs and/or anti-HBc antibodies) and occult HBV infection (positivity for HBV-DNA by nested PCR) in the serum of anti-HCV-positive, HCV-RNA-positive, HBsAg-negative patients with various degrees of CLD seen at a tertiary referral centre. PATIENTS: A total of 119 patients fulfilled the inclusion criteria (84 chronic hepatitis and 35 liver cirrhosis). RESULTS: Forty-eight patients (40.3%) showed markers of previous HBV infection. This feature was more frequent (P = 0.02) among cirrhotics (57%) as compared to chronic hepatitis patients (33%). Chronic hepatitis patients positive for markers of previous HBV infection had worse histology as compared to negative ones (grading: 6.4 +/- 2.7 versus 4.6 +/- 3.0, P = 0.004; staging: 1.6 +/- 1.2 versus 1.0 +/- 1.0, P = 0.01). Eight patients were positive for HBV-DNA in serum (6.7%). No difference in the presence of occult HBV infection was seen between various degrees of liver disease (7.1% of chronic hepatitis, 5.7% of cirrhosis) and among patients who were positive (10.4%) or negative (4.2%) for markers of previous HBV infection. No significant biochemical, virological, or histological difference was observed between age, age at infection, duration of infection, marker patterns of previous HBV infection-matched HBV-DNA-positive and negative chronic hepatitis patients. CONCLUSIONS: Our findings suggest that previous HBV infection among anti-HCV patients is associated with worse disease stage. In these patients, the prevalence of occult HBV infection is low and there is no difference in distribution among patients with or without markers of previous HBV infection. Furthermore, it does not seem to be associated with disease stage. Lastly, at least among patients with chronic hepatitis, it does not seem to affect the severity of disease. PMID- 12640724 TI - Quantitative versus morphological assessment of liver fibrosis: semi-quantitative scores are more robust than digital image fibrosis area estimation. AB - BACKGROUND/AIM: Digital image analysis (DIA) allows quantitative assessment of fibrosis on liver biopsy. Accurate determination of a threshold greyscale level representing fibrous tissue is critical. This method has not been fully evaluated in clinical practice. METHODS: Digital images of stained liver biopsy sections were captured by microscopy and converted to greyscale. A novel method of determining the threshold greyscale value at which to measure fibrosis area was developed (peak proportion area change (PPAC)). Reproducibility was tested in comparison with standard interactive thresholding and with semi-quantitative scoring using the Histological activity index (HAI) system by a histopathologist. Fibrosis areas for different sections from the same biopsy core were also compared by each method. RESULTS: Comparison between PPAC and interactive thresholding method demonstrated superior reproducibility of the PPAC method: r > 0.7, P < 0.001 compared with r = 0.19-0.64 (not all reaching significance). On a single section, reproducibility was similar for PPAC and the modified HAI system. When different sections from the same core were compared, the HAI system was more robust. CONCLUSIONS: The PPAC method is superior to standard interactive thresholding. However, variability in DIA scores between sections invalidates the technique for clinical use and semi-quantitative scoring systems remain the gold standard for fibrosis assessment. PMID- 12640725 TI - Hepatocellular adenoma and polycystic ovary syndrome. AB - Various identified risk factors predispose to hepatocellular adenomas. We present the case of a young woman with liver adenoma in a context of polycystic ovary syndrome, associated with high levels of androgens and following a high dose hormonal therapy. In view of this complication, we recommend a close screening of patients with such hormonal imbalance, especially those who are treated with high doses of hormones, with repeated liver tests and ultrasonographies. PMID- 12640726 TI - Des-gamma-carboxy prothrombin (DCP) ratio, a novel parameter measured by monoclonal antibodies MU-3 and 19B7, as a new prognostic indicator for hepatocellular carcinoma. AB - BACKGROUND/AIM: Recently, a novel des-gamma-carboxy prothrombin (DCP) antibody named 19B7 has been developed. It has been reported that the DCP ratio defined by DCP measured by MU-3 antibody/DCP measured by 19B7 antibody demonstrated differences between hepatocellular carcinoma and vitamin K deficiency. The aim of this study was to clarify the relationship between the DCP ratio and tumour malignancy, as well as the prognostic significance of the DCP ratio after hepatectomy for hepatocellular carcinoma. METHODS: From January 1996 to December 1999, 79 patients diagnosed with hepatocellular carcinoma underwent hepatectomy. To detect DCP in the plasma before surgery, we used a new EIA kit, Eitest PIVKA II kit (Eisai, Tokyo, Japan) with both MU-3 and 19B7 antibody. The DCP ratio was calculated using the formula: DCP ratio = DCP level measured by MU-3 antibody (mAU/ml)/DCP level measured by 19B7 antibody (mAU/ml). Pathologic findings using resected specimens were classified according to the Japanese General Rules for Primary Liver Cancer. RESULTS: Among DCP-positive patients, the DCP ratio was significantly higher in those with tumour thrombus in the portal vein (vp) or with a moderately or poorly differentiated hepatocellular carcinoma than in those without vp or with a well-differentiated carcinoma (P = 0.0162, 0.0109, respectively). Patients with DCP ratio > 3 had a significantly higher incidence of vp than those with DCP ratio < or = 3 (P = 0.0027). The survival rate was significantly higher in patients with DCP ratio < or = 3 than in those with DCP ratio > 3 (P = 0.0002). This result was the same as in DCP positive cases (P = 0.0027). CONCLUSIONS: The DCP ratio can distinguish DCP produced by hepatoma cells from DCP produced by normal hepatocytes in vitamin K deficiency. The DCP ratio is a useful prognostic indicator that can be assessed before hepatectomy. PMID- 12640727 TI - Five-year survival predictive factors in patients with excessive alcohol intake and cirrhosis. Effect of alcoholic hepatitis, smoking and abstinence. AB - AIM: To evaluate 5-year survival predictive factors in hospitalised patients with excessive alcohol intake and cirrhosis, including in a multivariate analysis the severity of the liver disease, gastrointestinal bleeding, concomitant viral B or C infection, smoking status, presence of alcoholic hepatitis at inclusion and abstinence from alcohol during follow-up. METHODS: In a non-concurrent cohort study, 122 patients with excessive alcohol intake and cirrhosis were followed up at least five years or till death. Two patients were lost to follow-up. RESULTS: The 5-year survival rates were 43% in the 122 patients and 66%, 50% and 25% in Child-Pugh class A, B and C patients, respectively. In multivariate analysis, age (P = 0.01), Child-Pugh score (P = 0.0001), gastrointestinal bleeding (P = 0.01), presence of HBs Ag and/or anti-HCV (P = 0.03), smoking (P = 0.01), absence of histologically proven alcoholic hepatitis (P = 0.05) and persistent alcohol intake (P = 0.002) were associated with significantly increased risk ratios of death. CONCLUSIONS: In hospitalised patients with excessive alcohol intake and cirrhosis: (1) age, liver failure, gastrointestinal bleeding, concomitant viral B or C infection and persistent alcohol intake are independent poor prognostic markers, (2) smoking may contribute to the aggravation of cirrhosis, and (3) alcoholic hepatitis, being a potentially reversible cause of liver failure, has a favourable prognostic significance. PMID- 12640728 TI - Portal vein arterialisation as a technical option in liver transplantation: impact on function, regeneration, and morphology of the liver following hemihepatectomy in pigs. AB - BACKGROUND: Arterialisation of the portal vein has been propagated as a technical variant in liver transplantation. However, the consequences of this unphysiological vascular supply are insufficiently known. METHODS: Twenty-seven healthy pigs were subjected to a left hemihepatectomy and divided into three groups. The first group received complete arterialisation of the portal vein (PVA). In the second group hepatic artery blood flow was also interrupted by dividing the artery (PVA/DHA). Nine animals served as controls (Group C). RESULTS: Early mortality in the PVA/DHA group (6/9) was significantly increased in comparison with the PVA (2/9) and control (2/9) groups (P < 0.05). In the surviving animals, arterialisation (PVA and PVA/DHA) led to significantly faster hepatic regeneration in comparison with control animals, with comparable liver function and with liver size increasing to 278% and 293% vs 134% (P = 0.002) after 3 weeks, in liver ... weeks. This was accompanied by enhanced hepatic expression of the proliferation markers MIB-1 (22.4% and 16.7% vs 5.9%, P = 0.002) and PCNA (86% and 68% vs 66%, P = 0.002) one week postoperation. At the same time, the number of apoptotic hepatocytes increased from 1.6% to 2.5% and 2.3% (P = 0.002). No significant difference was found in the collagen content of the liver after 3 weeks. CONCLUSIONS: Arterialisation of the portal vein promotes early and enhanced hepatic regeneration without impairing liver function. This technique may therefore be useful in split-graft liver transplantation, where this aspect would be of particular importance. PMID- 12640729 TI - Impact of oestrogens on the progression of liver disease. AB - As of this writing, the most common cause of hepatic fibrosis is chronic hepatitis C virus infection (HCV), the characteristic feature of which is hepatic steatosis. Hepatic steatosis leads to an increase in lipid peroxidation in hepatocytes, which in turn activates hepatic stellate cells (HSCs). HSCs are also thought to be the primary target cells for inflammatory stimuli, and produce extracellular matrix components. Based on available clinical information, chronic hepatitis C appears to progress more rapidly in men than in women, and cirrhosis is predominately a disease of men and postmenopausal women. It should be noted that estradiol (E2) is a potent endogenous antioxidant. A recent study has shown that hepatic steatosis became evident in an aromatase-deficient mouse and was diminished in animals, after treatment with E2. Our studies showed that E2 suppressed hepatic fibrosis in hepatic fibrosis models, inhibited the activation of activator protein 1 and nuclear factor-kappa B in cultured hepatocytes undergoing oxidative stress, and attenuated HSC activation in primary culture. Recently, variant oestrogen receptors (ERs) were found to be expressed to a greater extent in male patients with chronic liver disease than in female subjects. We also demonstrated decreased levels of ERs in postmenopausal women and cirrhotic patients of both genders. The actions of E2 are mediated through ER alpha and beta. HSCs have also been found to possess functional ER beta but not ER alpha. A better understanding the basic mechanisms underlying the gender associated differences observed in the development of hepatic fibrosis would provide valuable information relative to the search for effective antifibrogenic therapies. PMID- 12640730 TI - Neural crest and tooth morphogenesis. AB - Mammalian teeth develop from two types of cells: stomodeal ectoderm, which forms ameloblasts, and cranial neural-crest-derived (ecto) mesenchyme cells, which form odontoblasts and cementoblasts. These two cell types, juxtaposed in the developing oral cavity, interact to control the entire process of tooth initiation, morphogenesis, and cytodifferentiation. Cell-cell signaling pathways and their target nuclear factors have been identified as key mediators of the progressively complex exchange of information between ectoderm and ectomesenchyme. The constantly changing direction of the reciprocal signaling and cell responses between ectoderm and ectomesenchyme enables cells to monitor their relative spatial positions and differentiated states continuously. The least understood of the early processes in tooth development are morphogenesis and patterning. From a seemingly uniform layer of oral ectoderm and underlying mass of ectomesenchyme, different types (shapes) of teeth develop in different positions. Tooth type is determined very early in development, before the overt onset of morphogenesis. Thus, the early ectoderm-ectomesenchyme cell interactions must in some way either create or respond to positional differences in the jaw primordia. PMID- 12640732 TI - Enamel knots as signaling centers linking tooth morphogenesis and odontoblast differentiation. AB - Odontoblasts differentiate from the cells of the dental papilla, and it has been well-established that their differentiation in developing teeth is induced by the dental epithelium. In experimental studies, no other mesenchymal cells have been shown to have the capacity to differentiate into odontoblasts, indicating that the dental papilla cells have been committed to odontoblast cell lineage during earlier developmental stages. We propose that the advancing differentiation within the odontoblast cell lineage is regulated by sequential epithelial signals. The first epithelial signals from the early oral ectoderm induce the odontogenic potential in the cranial neural crest cells. The next step in the determination of the odontogenic cell lineage is the development of the dental papilla from odontogenic mesenchyme. The formation of the dental papilla starts at the onset of the transition from the bud to the cap stage of tooth morphogenesis, and this is regulated by epithelial signals from the primary enamel knot. The primary enamel knot is a signaling center which forms at the tip of the epithelial tooth bud. It becomes fully developed and morphologically discernible in the cap-stage dental epithelium and expresses at least ten different signaling molecules belonging to the BMP, FGF, Hh, and Wnt families. In molar teeth, secondary enamel knots appear in the enamel epithelium at the sites of the future cusps. They also express several signaling molecules, and their formation precedes the folding and growth of the epithelium. The differentiation of odontoblasts always starts from the tips of the cusps, and therefore, it is conceivable that some of the signals expressed in the enamel knots may act as inducers of odontoblast differentiation. The functions of the different signals in enamel knots are not precisely known. We have shown that FGFs stimulate the proliferation of mesenchymal as well as epithelial cells, and they may also regulate the growth of the cusps. We have proposed that the enamel knot signals also have important roles, together with mesenchymal signals, in regulating the patterning of the cusps and hence the shape of the tooth crown. We suggest that the enamel knots are central regulators of tooth development, since they link cell differentiation to morphogenesis. PMID- 12640731 TI - Epigenetic signals during odontoblast differentiation. AB - Odontoblast terminal differentiation occurs according to a tooth-specific pattern and implies both temporospatially regulated epigenetic signaling and the expression of specific competence. Differentiation of odontoblasts (withdrawal from the cell cycle, cytological polarization, and secretion of predentin/dentin) is controlled by the inner dental epithelium, and the basement membrane (BM) plays a major role both as a substrate and as a reservoir of paracrine molecules. Cytological differentiation implies changes in the organization of the cytoskeleton and is controlled by cytoskeleton-plasma membrane-extracellular matrix interactions. Fibronectin is re-distributed during odontoblast polarization and interacts with cell-surface molecules. A non-integrin 165-kDa fibronectin-binding protein, transiently expressed by odontoblasts, is involved in microfilament reorganization. Growth factors (TGF beta 1, 2, 3/BMP2, 4, and 6), expressed in tooth germs, signal differentiation. Systemically derived molecules (IGF1) may also intervene. IGF1 stimulates cytological but not functional differentiation of odontoblasts: The two events can thus be separated. Immobilized TGF beta 1 (combined with heparin) induced odontoblast differentiation. Only immobilized TGF beta 1 and 3 or a combination of FGF1 and TGF beta 1 stimulated the differentiation of functional odontoblasts over extended areas and allowed for maintenance of gradients of differentiation. Presentation of active molecules in vitro appeared to be of major importance; the BM should fulfill this role in vivo by immobilizing and spatially presenting TGF beta s. Attempts are being made to investigate the mechanisms which spatially control the initiation of odontoblast differentiation and those which regulate its propagation. Analysis of molar development suggested that odontoblast differentiation and crown morphogenesis are interdependent, although the possibility of co-regulation requires further investigation. PMID- 12640733 TI - Molecular insights into the lineage-specific determination of odontoblasts: the role of Cbfa1. AB - The role of stable transcription complexes in initiating and consolidating programs of gene expression during lineage specification has been extensively studied. Despite the progress made in the identification of key molecules of tooth initiation and patterning, the mechanisms leading to cell differentiation during odontogenesis are unknown. Odontoblasts are exclusive dentin-producing cells that are phenotypically and functionally distinct from osteoblasts. However, not much is known about the precise determinants of odontoblast terminal differentiation--in particular, how the fate of these cells becomes delineated from that of osteogenic mesenchyme. Cbfa1(-/-) mice completely lack osteoblasts and bone, while tooth development arrests at the time of odontoblast differentiation. The purpose of this paper is to overview our studies on the role of Cbfa1 in odontoblast determination and differentiation using the Cbfa1(-/-) mouse model and various experimental approaches. Our expression analyses confirm the down-regulation of Cbfa1 expression in newly differentiated and functional odontoblasts. Second, we demonstrate that Cbfa1(-/-) incisor organs arrest at a later stage than molars, and that alpha 1 (I) collagen, a marker of odontoblast differentiation shared in common with osteoblasts, is not significantly affected by the absence of the transcription factor. Interestingly, Dspp expression in Cbfa1(-/-) appeared markedly down-regulated in putative odontoblasts. The overexpression of Cbfa1 in an odontoblast cell line (MDPC-23) results in the selective down-regulation of Dspp and not type I collagen. It is likely that, in addition to its influence on tooth epithelial morphogenesis, Cbfa1 plays a non redundant and stage-specific role in the lineage determination and terminal differentiation of odontoblasts from dental papilla mesenchyme. PMID- 12640734 TI - Utilization of MO6-G3 immortalized odontoblast cells in studies regarding dentinogenesis. AB - Tooth formation is the result of reciprocal instructive interactions between oral epithelium and cranial neural-crest-derived ectomesenchymal tissues. These interactions lead to the cytodifferentiation of highly specialized matrix-forming cell types, the ameloblast, odontoblast, and cementoblast, that produce the mineralized tissues enamel, dentin, and cementum, respectively. Our laboratory has been developing immortalized dental cell lines representative of these various cell types to facilitate studies on gene regulation, cell differentiation, matrix formation, and mineralization. Odontoblasts are solely responsible for the synthesis and secretion of the dentin extracellular matrix bilayer that consists of non-mineralized predentin and mineralized dentin. The mouse immortalized MO6-G3 cell line expresses the major matrix proteins associated with the odontoblast phenotype, producing a matrix that is capable of mineralization. This cell line serves as a useful tool in studies designed to explore the various processes of dentinogenesis. In this paper, we present studies using the mouse odontoblast cell line MO6-G3 as examples of the various research applications. Studies highlighted are: in vitro promoter studies investigating the tooth-specific gene regulation of the major non-collagenous dentin matrix protein, dentin sialophosphoprotein; regulation of tertiary dentin formation by cytokines, such as transforming growth factor-Beta 1; and the utilization of dentally relevant cells in dental material biocompatibility testing. PMID- 12640735 TI - New genes involved in odontoblast differentiation. AB - The odontoblast phenotype has been mainly approached by the biochemical characterization of dentin matrix proteins and by extrapolation of the knowledge of bone cell biology, since dentin and bone share many similarities. In fact, direct investigations of the odontoblast phenotype have been hindered by the limited number of cells within the dental pulp and the difficulty in microdissection and isolation of a pure population of these cells. To overcome these obstacles, we previously developed a cell-culture system that promotes differentiation of human dental pulp cells into odontoblasts. This material now permits the study of odontoblasts through molecular biology techniques. Therefore, we constructed a cDNA library enriched for odontoblast-specific genes using the suppression subtractive hybridization technique (SSH). This library led us to identify new genes expressed by odontoblasts. In this paper, we will focus on some genes implied in various functions associated with odontoblast differentiation, such as cell polarization (MAP1B), dentin mineralization (PHEX, osteoadherin), and relationships between odontoblasts and nerve cells (reelin). Another important fact is that about 40% of the cDNA were unknown genes. Therefore, one can speculate that some of them will be odontoblast-specific, since, until now, only one gene (DSPP) presents this characteristic. PMID- 12640736 TI - FGFs-1 and -2, and TGF beta 1 as inductive signals modulating in vitro odontoblast differentiation. AB - We have studied the expression of FGF1 and FGF2 during mouse odontogenesis by immunohistochemistry. FGF1 was detected in differentiated odontoblasts and at the secretory pole of ameloblasts. Localization of FGF2 was mainly observed within the basement membrane interposed between dental epithelium and dental mesenchyme. These findings indicate that FGF1 and FGF2 may participate in the control of odontoblast and ameloblast differentiation. Thereafter, we studied the ability of FGF1 and FGF2, alone or in combination with TGF beta 1, to induce polarization and/or functional differentiation of preodontoblasts. Dental papillae (DP) obtained from first lower molars of 17-day-old mouse embryo were cultured in the presence or the absence of growth factors. DP cultured with FGF1 + TGF beta 1 showed gradients of odontoblast-like cell differentiation, which displayed alkaline phosphatase reactivity. DP treated with FGF2 + TGF beta 1 exhibited pre odontoblast cell polarization, and the cell bodies displayed long cytoplasm processes. However, following this treatment we did not observe extracellular matrix secretion, and alkaline phosphatase activity was completely inhibited. In summary, our results show that exogenous addition of FGF1 to pre-odontoblasts induces their terminal differentiation, by synergistically acting with TGF beta 1. In contrast, FGF2 may regulate the effect of TGF beta 1, permitting cell polarization but restraining pre-odontoblast functions. PMID- 12640737 TI - The effect of TGF-beta 2 on dentin apposition and hardness in transgenic mice. AB - Transforming growth factor beta, TGF-beta, is expressed during tooth formation and can induce pre-odontoblast differentiation and formation of functional odontoblast-like cells in vitro. In addition, exogenous TGF-beta can increase reparative dentin formation, presumably by acting on odontoblasts. In this study, we examined the tooth phenotype of transgenic mice, in which TGF-beta 2 expression is directed by the osteocalcin promoter. Previous studies have shown that these mice have a bone phenotype that resembles that of human osteoporosis, including the existence of spontaneous fractures. Microhardness testing of the enamel and dentin showed no differences in the molars of these transgenic mice as compared with those of their wild-type littermates. Consistent with the increase in bone mineral apposition rate previously reported in these mice, the dentin apposition rate appeared to be increased in the TGF-beta 2-overexpressing mice. Thus, in teeth, as in bone, TGF-beta 2 appears to stimulate the synthesis and deposition of matrix. Further studies are needed to understand the effect of TGF beta 2 on distinct mineralized tissues (bone, dentin, and cementum) and to determine whether exogenous TGF-beta 2 may be useful for tooth repair. PMID- 12640738 TI - Sex- and age-related differences in primary and secondary dentin formation. AB - Clinical studies carried out on dentin thickness in adults, as well as experimental studies carried out on ovariectomized animals, indicate that odontoblast activity, like that of osteoblastic cells, differs in the two sexes. To examine the evidence for differences in odontoblast activity before puberty, we have measured dentin thickness and other crown dimensions from bitewing radiographs of the lower first molars in 240 children aged 4-16 years. The radiographs were obtained from pedodontic clinics throughout Israel. Only teeth without caries or fillings were used, and the study population had minimal attrition. The results showed that dentin thickness, measured on the roof of the pulp chamber, was significantly greater in boys than in girls at all ages, and that the differences increased during puberty. The differences remained highly significant even when standardized for crown size. They demonstrate that dimorphism in dentin thickness is present even in the earliest stages of odontogenesis and increase with puberty. PMID- 12640739 TI - Molecular regulation of odontoblast activity under dentin injury. AB - Pulp tissue responds to dentin damage by laying down a tertiary dentin matrix (reactionary or reparative) beneath the site of injury. Reactionary dentin is secreted by surviving odontoblasts in response to environmental stimuli, leading to an increase in metabolic activities of the cells. The inductive molecules that determine the success of the pulp healing may be released from the damaged dentin as well as from the pulp tissue subjacent to the injury. This paper will schematically consider two major growth factors probably implicated in the control of odontoblast activity: TGF beta-1 released from demineralized dentin and NGF from pulp. To analyze their role with an in vitro system that mimics the in vivo situation, we have used thick-sliced teeth cultured as described previously. The supply of factors was accomplished by means of a small tube glued onto the dentin. The tube was filled with TGF beta-1 (20 ng/mL) or NGF (50 ng/mL), and slices were cultured for 4 or 7 days. Results showed that TGF beta-1 binding sites are strongly detected on odontoblasts in the factor-rich zone. A strong expression of alpha 1(I) collagen transcripts was also detected. In the NGF-rich environment, p75NTR was re-expressed on odontoblasts and the transcription factor NF-kappa B activated. Modifications in the odontoblast morphology were observed with an atypical extension of the cell processes filled with actin filaments. These results suggest that odontoblasts respond to influences from both dentin and pulp tissue during pulp repair. PMID- 12640740 TI - Trans-dentinal stimulation of tertiary dentinogenesis. AB - Trans-dentinal stimulation of tertiary dentinogenesis has long been recognized, and has traditionally been ascribed to diffusion of irritant substances arising during injury and restorative treatment. Identification of bio-active components, especially growth factors including TGF-beta s, sequestered within dentin matrix provides a new explanation for cellular signaling during tertiary dentinogenesis. Both isolated dentin matrix components and pure growth factors (TGF-beta s) have been shown to signal cellular events leading to reactionary and reparative tertiary dentinogenesis. Release of these bio-active components from dentin matrix may arise during carious attack and other injury to the tissue, and also during subsequent surgical intervention and restoration of the tooth. Both cavity conditioning agents and leaching from restorative materials may contribute to release of these components. Distance of diffusion, as determined by cavity residual dentin thickness, and other restorative parameters may influence the signaling process after release of these components. Careful consideration of the interplay between tissue injury and surgical and restorative material factors is required for optimum exploitation of the exquisite regenerative capacity of dentin-pulp for more biological approaches to clinical treatment of dental disease. PMID- 12640741 TI - Human odontoblast culture method: the expression of collagen and matrix metalloproteinases (MMPs). AB - Studies on mature human odontoblasts have suffered for the lack of in vitro models. We recently introduced a human odontoblast and pulp tissue organ culture method, in which the odontoblasts are cultured in the pulp chamber after removal of the pulp tissue, and the pulp tissue can be cultured separately (Tjaderhane et al., 1998a). With this method, we have studied the effects of growth factors on the expression of collagen and extracellular matrix (ECM)-degrading enzymes, matrix metalloproteinases (MMPs), in mature human odontoblasts. TGF-beta 1 was selected because of its ability to regulate the response of the dentin-pulp complex to external irritation. The effect of TGF-beta 1 (10 ng/mL) on pro alpha 1(I) collagen mRNA was analyzed by quantitative PCR, and type I procollagen propeptide (PINP) was analyzed from conditioned culture media with RIA. Odontoblast media were also assayed for respective type III procollagen propeptide (PIIINP). TGF-beta had a negligible effect on collagen mRNA expression or protein synthesis, indicating that TGF-beta alone does not markedly induce dentin matrix formation per se in the human dentin-pulp complex (Palosaari et al., 2001). However, TGF-beta 1 seems to regulate MMP expression in mature human odontoblasts differentially. A strong down-regulation of MMP-8 (Palosaari et al., 2000), a modest down-regulation of MMP-20 (Tjaderhane et al., 2000), and considerable up-regulation of MMP-9, with no apparent effect on MMP-2 expression (Tjaderhane et al., 1998b), indicate that growth factors may affect the matrix synthesis by controlling the expression and activity of MMPs instead of collagen synthesis. The altered expression of MMPs may result in altered ECM formation, which in turn may contribute to the formation of atubular reparative dentin. PMID- 12640742 TI - Molecular aspects of tooth pathogenesis and repair: in vivo and in vitro models. AB - Several growth factors and extracellular matrix molecules, which are expressed during embryonic tooth development, are re-expressed in dental tissues under pathological conditions. Pathological conditions such as caries lesions and dental injuries are often lethal to the odontoblasts, which are then replaced by other pulp cells. These cells are able to differentiate into odontoblast-like cells and produce a reparative dentin. Here we demonstrate the in vivo distribution of several molecules in human permanent teeth under normal and pathological conditions. The intermediate filament protein nestin, which is a marker of young odontoblasts, is absent from old permanent teeth. Similarly, the Notch protein, which is involved in cell fate specification and is localized in the sub-odontoblastic cell layer during odontogenesis, is not detected in adult dental tissues. In carious and injured teeth, nestin is expressed in a selective manner in odontoblasts surrounding the injury site, while Notch is expressed in the sub-odontoblastic layer of cells. We reproduced this physiological event in an in vitro culture system. Pulp cells cultured in the presence of beta glycerophosphate formed mineralization nodules. As odontoblasts, pulp cells contributing to the nodule formation express type I collagen, osteonectin, dentin sialophosphoprotein, and nestin. In this in vitro assay system, nestin is up regulated after local application of Bone Morphogenetic Protein 2 and 4. Fourier transform infrared microspectroscopy showed that both the organic and the mineral compositions of the nodules have the characteristics of human dentin and differ from those of enamel and bone. These findings show that both the molecular and the mineral characteristics of the human dentin matrix are respected in the in vitro culture conditions. PMID- 12640743 TI - Expression of TGF-beta receptors I and II in the human dental pulp by in situ hybridization. AB - Members of the TGF-beta family of growth factors are important in modulation of odontoblast secretory activity during dental tissue repair. Odontoblast expression of TGF-beta isoforms during development leads to their sequestration within the dentin matrix, from where they may be released during carious injury and participate in reparative processes. Two receptors, implicated in TGF-beta mediated cell signaling, have been identified immunohistochemically in both odontoblasts and pulpal cells of healthy and carious human molar teeth. This study aimed to characterize the expression of the TGF-beta receptors I and II in sound and carious teeth by means of in situ hybridization, to help our understanding of the response of these cells to TGF-beta stimulation. Sound and carious human third molar teeth were routinely processed immediately following extraction, and 10-microns paraffin-embedded sections prepared. These sections were hybridized with 32P-labeled probes to TGF-beta receptors I and II, and the subsequent signal was detected by autoradiography. mRNA for both receptors I and II was mainly detected within the odontoblasts and nerve-associated cells of healthy tissues, with expression at lower levels seen within the subodontoblast and pulp core cells. The expression in odontoblasts was higher for TGF-beta receptor I than for receptor II. Expression of both receptors was more homogenous in all pulp cells within carious teeth, because of an increase of signal within the underlying pulp cell population, including blood-vessel-associated cells. We conclude that the TGF-beta receptors I and II were expressed in odontoblasts and pulp cells, and that subtle variations in the levels of their expression could be involved in the tissue response to injury. PMID- 12640744 TI - Odontoblast function seen as the response of dentinal tissue to dental caries. AB - Microbes are responsible for the initiation and maintaining of carious processes. They have an efficient machinery for dissolving crystalline hydroxyapatite. When initiating carious processes, microbial acid formation determines the rate of the process in enamel. When the process reaches dentin, the micro-environment changes. Dential fluid in dentin tubules is the liquid where dissolving products of apatites are destroyed. Inorganic composition of dentinal fluid, however, is not altered much during the carious process, indicating that a functional secretory domain is working to pump the dissolved calcium and phosphate ions out of the fluid. Activation of odontoblast alkaline phosphatase and dentin latent collagenases is the known cellular event during the carious process in dentin. Because the caries lesion is by definition undermining, this suggests that, in this degradation process, the extracellular compartment, crystalline hydroxyapatite is dissolved by microbial acids, and a mixture of proteinases degrades the organic matrix. The degradation products of collagen and other matrix components in dentinal fluid must be transported either through the caries lesion in the enamel to saliva or through the odontoblast to the pulp (active transport). This facilitates further processing of the degradation products intracellularly during the passage through the cell. PMID- 12640745 TI - The effects of high levels of glucose and insulin on type I collagen synthesis in mature human odontoblasts and pulp tissue in vitro. AB - High levels of dietary sucrose affect the metabolism of the pulp-dentin complex and enhance the caries process in dentin. The high-sucrose diet reduces dentin formation in young rats (Tjaderhane et al., 1994; Hietala and Larmas, 1995; Tjaderhane, 1996) and in pups of rat dams fed high-sucrose diet during lactation (Pekkala et al., 2000a). However, the mechanisms behind the effects are unknown. A direct effect of elevated blood glucose or an indirect effect via insulin has been suggested. We investigated the effects of high glucose and insulin on type I collagen synthesis in human odontoblasts and pulp tissue in vitro, using an organ culture method for functional post-mitotic odontoblasts. Odontoblasts and pulp tissue were cultured separately for 10 days in DMEM with 15% FBS containing additional glucose (G) (4.45 g/L) or insulin (I) (0.6 microgram/mL) or both together (GI). We evaluated type I collagen synthesis with RIA, measuring the level of N-terminal propeptide of type I collagen (PINP) secreted into the culture media. PINP secretion decreased in odontoblasts and pulp tissue in G and GI groups when compared with the control and insulin samples (p = 0.001 in both groups in the pulp samples). Insulin alone did not affect PINP secretion distinctly. The results indicate that high levels of glucose, but not insulin, directly down-regulate the type I collagen synthesis in young, differentiated human odontoblasts and pulp tissue. Insulin does not affect the inhibitory effect of high sucrose. These in vitro findings indicate that the high-sucrose diet may alter odontoblast function independently of insulin. PMID- 12640746 TI - Further observations on tertiary dentin in human deciduous teeth. AB - The structure of reparative tertiary dentin in human deciduous teeth has been studied. Reparative dentin is secreted by a new generation of odontoblast-like cells which have been subject to strong stimuli, e.g., trauma or deep active caries lesions with associated pulp inflammation. Ground sections of 25 teeth were prepared, and contact microradiographs were produced. Another 30 teeth were demineralized, embedded in paraffin, sectioned, and stained with hematoxylin and eosin. Some demineralized sections from each tooth were also studied in the scanning electron microscope. Most of the teeth showed some type of tertiary dentin formation. Mineralized tissue with a varied morphology was observed. In teeth which had been subject to trauma, the entire pulp chamber was sometimes obliterated. Mineralization seemed to start in the incisal region, and the central part of the pulp was the last part to be obliterated. Radiolucent voids and canals were seen. The organic matrix was dense and fibrous. In the pulp chamber and especially in the root canals, resorption had often occurred, indicating that signals giving rise to odontoclasts were also present. Resorption was often followed by deposition of various amounts of cementum-like repair tissue. The cells responsible for the formation of reparative dentin are believed to be subodontoblasts or undifferentiated ectomesenchymal cells. The varied morphology of the reparative dentin, observed in the pulp of the teeth examined, indicates that different stimuli lead to induction of hard-tissue-forming cells which produce different types of hard tissue. PMID- 12640747 TI - Presence or absence of tertiary dentinogenesis in relation to caries progression. AB - Studies have shown that dental caries may or may not be associated with tertiary dentin formation in the pulp. On the basis of histological examinations of 69 clinical well-defined caries lesions, a hypothesis is proposed on the dynamics of the hard-tissue responses of the pulp to caries. In active non-cavitated lesions, the formation of tertiary dentin seems to be initiated by primary odontoblast cells that subsequently result in atubular dentin/fibrodentinogenesis, whereas, in similarly aged but more rapidly progressing cavitated enamel lesions, no tertiary dentin is laid down by primary odontoblast cells. In all old-dentin exposed lesions, a so-called closed lesion environment was defined with subjacent atubular dentin formation. As these lesions progress, a shift from a closed to a more large and open lesion environment may develop in the very old lesions, and a new tubular dentinal matrix is noted on the top of the fibrodentin, also defined as reparative dentinogenesis. In very old slowly progressing lesions, a relatively small open lesion environment is also observed, with tubular tertiary dentin resembling the primary dentin being strictly tubular. It is suggested that the absence of tertiary dentinogenesis can be expected in very rapid caries lesions, whereas a variety of tertiary dentin is observed in older dentin cavitated lesions guided by a changing external lesion environment over time. PMID- 12640748 TI - Factors in pulpal repair after oral exposure. PMID- 12640749 TI - Technique sensitivity: biological factors contributing to clinical success with various restorative materials. AB - Since the 1950s, clinicians have relied on various formulations of Ca(OH)2 to stimulate dentin bridge formation. Various studies (Kozlov and Massler, 1966; Massler, 1967; Brannstrom, 1978; Cox et al., 1987; Snuggs et al., 1993) have demonstrated that pulp healing and dentin bridging can occur against a pH spectrum of materials. Recent studies (Akimoto et al., 1998; Cox et al., 1998, 1999; Tarim et al., 1998; Kitasako et al., 1999; Hafez et al., 2000) have reported successful pulp healing and dentin bridging using adhesives for direct capping of exposed pulps. However, others (Costa et al., 1997; Stanley and Pameijer, 1997; Pameijer, 1998; Hebling et al., 1999; Carvalho et al., 2000) have reported unsatisfactory results when exposures were direct-capped with adhesives. Biological and technical factors, or a combination of both, might be postulated to explain these differences. Recent studies have demonstrated that biological success is dependent upon proper hemorrhage control at the exposure site. This review explores the differences and common factors influencing successful dentin bridging, focusing on data derived from animal studies conducted according to ISO usage guidelines for cavity preparation and material placement. In the past, there has been concern that etching of vital dentin leads to immediate pulp death due to low pH. Recent studies have reported that acidic cements cause breakdown of only the smear layer and fail to seal the restoration interface, leading to inflammation and necrosis. A properly hybridized dentinadhesive interface provides a "bacteriometic" seal to both dentin and pulp tissues. Recent ISO usage studies have shown a high incidence of dentin bridging with adhesives following proper hemorrhage control and removal of both operative debris and biofilm at the dentin-pulp interface by agents such as NaOCl. These are important technique sensitivity factors to be considered for pulp healing and dentin bridge formation. PMID- 12640750 TI - Application of bioactive molecules in pulp-capping situations. AB - To evaluate the effects of bioactive molecules in pulpal wound healing, we carried out experiments using the rat upper molars as an in vivo model. Cavities were prepared on the mesial aspect, and pulp perforation was accomplished by the application of pressure with the tip of a steel probe. After the pulp-capping procedure, the cavities were filled with a glass-ionomer cement. Comparison was made between and among: (1) sham-operated controls with dentin and predentin fragments implanted in the pulp during perforation after 8, 14, and 28 days; (2) carrier without bioactive substance; (3) calcium hydroxide; (4) Bone Sialoprotein (BSP); (5) different concentrations of Bone Morphogenetic Protein-7 (BMP-7), also termed Osteogenic Protein-1 (OP-1); and (6) N-Acetyl Cysteine (NAC), an anti oxidant agent preventing glutathione depletion. Histologic and morphometric comparison, carried out among the first 4 groups on demineralized tissue sections, indicated that, at 28 days after implantation, BSP was the most efficient bioactive molecule, inducing homogeneous and well-mineralized reparative dentin. BMP-7 gave reparative dentin of the osteodentin type in the coronal part of the pulp, and generated the formation of a homogeneous mineralized structure in the root canal. These findings indicate that the crown and radicular parts of the pulp bear their own specificity. Both BSP and BMP-7 were superior to calcium hydroxide in their mineralization-inducing properties, and displayed larger areas of mineralization containing fewer pulp tissue inclusions. The overall mineralization process to these molecules appeared to proceed by mechanisms that involved the recruitment of cells which differentiate into osteoblast-like cells, producing a mineralizing extracellular matrix. We also provide preliminary evidence that NAC induces reparative dentin formation in the rat molar model. Pulp-capping with bioactive molecules provides new prospects for dental therapy. PMID- 12640751 TI - Dentin regeneration in vital pulp therapy: design principles. AB - The nature and specificity of the mechanisms by which the amputated dentin-pulp interface is therapeutically healed determine the properties of the barrier at this site and play a critical role in the outcome of vital pulp therapy. Healing of the dentin-pulp complex proceeds either by natural repair-which results in defensive hard-tissue formation, or therapeutically regulated dentin regeneration, which aims to reconstitute the normal tissue architecture at the pulp periphery. Progress in biomedical research opens new directions for the design of biologically effective pulp therapies. Application of biocompatible and biodegradable carrier vehicles for local delivery of signaling molecules in pulp capping situations showed induction of fibrodentin/reparative dentin formation, but often at the expense of underlying pulp tissue. An alternative pre-clinical model aiming to reconstitute normal tissue architecture directly at the dentin pulp interface should be designed on the basis of the direct induction of odontoblast-like cell differentiation and reparative dentin formation at the pulp capping material interface. Experimental data clearly showed that pulpal cells can differentiate directly into odontoblast-like cells in association with specific extracellular matrices (dentinal or fibrodentinal matrix) or TGF beta 1 containing artificial substrates. Dentin-induced dentinogenesis can be used as a master plan for the achievement of new therapeutic opportunities. In the present study, several short-term experimental studies on dog teeth for potential direct induction of odontoblast-like cell differentiation at the surface of rhTGF beta 1 containing artificial substrates (Millipore filters, hydroxyapatite granules, calcium hydroxide, pure titanium) failed to induce any specific reparative dentinogenic effects. PMID- 12640752 TI - The in vitro and in vivo influence of 4-META/MMA-TBB resin components on dental pulp tissues. AB - The purpose of this study was to qualitate the penetration of the major components of 4-META/MMA-TBB adhesive resin (4-META resin) and to characterize their influence on the in vitro and in vivo wound healing of dental pulp tissues. Fresh 4-META resin was applied to rabbit mesentery; its components penetrated the mesentery to form three of layers, depending on the amounts of monomer components in the tissue. The superficial layer was a soft-tissue hybrid layer (STHL), the intermediate layer contained small particles of polymerized 4-META resin, while the deepest layer contained unpolymerized monomer components including MMA and butanol, which were detected by gas chromatography (GC). To characterize the in vivo effects of the deepest layer, we immersed the pulp tissue in MMA or in 5% 4 META/MMA and autotransplanted it to placement beneath a rabbit kidney capsule. The MMA-immersed pulp was positive for osteocalcin and presented osteodentin formation at 7 days, as did the untreated control pulp tissue. In contrast, the 5% 4-META/MMA-immersed pulp collapsed into the cell-deficient fibrous connective tissue, with slight calcification by 7 days and less osteodentin formation at 14 days. Analysis of these data suggests that MMA does not inhibit osteogenic activity of pulp tissue, while 5% 4-META/MMA does inhibit osteogenic activity to some extent. PMID- 12640753 TI - Enamel matrix derivative promotes reparative processes in the dental pulp. AB - During odontogenesis, amelogenins from the preameloblasts are translocated to differentiating odontoblasts in the dental papilla, suggesting that amelogenins may be associated with odontoblast changes during development. In the present study, we have explored the effects of enamel matrix derivative (EMD) on the healing of a pulpal wound. Coronal pulp tissue of permanent maxillary premolars of miniature swine were exposed through buccal class V cavities. The exposed pulp was capped with EMD. The contralateral teeth served as controls and were capped with a calcium hydroxide paste (Dycal). The cavities were sealed with glass ionomer cement. After 2 and 4 weeks, the histology of the teeth was analyzed. In the EMD-treated teeth, large amounts of newly formed dentin-like hard tissue with associated formative cells outlined the pulpal wound separating the cavity area from the remaining pulp tissue. Inflammatory cells were present in the wound area but not subjacent to the newly formed hard tissue. Morphometric analysis showed that the amount of hard tissue formed in EMD-treated teeth was more than twice that of the calcium-hydroxide-treated control teeth (p < 0.001), suggesting that EMD is capable of promoting reparative processes in the wounded pulp more strongly than is calcium hydroxide. PMID- 12640754 TI - Pulp-capping with recombinant human insulin-like growth factor I (rhIGF-I) in rat molars. AB - The aim of this study was to explore pulp healing and reparative dentinogenesis following pulp-capping by using recombinant human insulin-like growth factor I (rhIGF-I). Exposures were made through the mesial pulp horn in first upper molars in two-month-old Wistar rats. The pulp was covered with one dose of sterile 4% methylcellulose gel containing either 400 ng rhIGF-I or saline in contralateral controls. The exposure site was closed with sterile Teflon membrane, and the cavity was filled with IRM cement. Additional molars were capped with Dycal as controls. After 3, 7, or 28 days, animals were anesthetized and fixed by intravascular glutaraldehyde perfusion. Molars were decalcified and processed for histological analysis and cut with membrane and residual methacrylate from IRM in situ. Only specimens with acceptable pulp sealing according to blinded microscopy control were included. On day 3, identical inflammatory responses in the upper pulp were observed in molars with rhIGF-I gel or control gel. On day 7, granulation tissue ingrowth had partly replaced inflammatory infiltration in both groups. After 28 days, complete dentin bridging and tubular dentin formation were observed more frequently and closer to the test substance containing rhIGF-I. The reparative dentin response to capping with rhIGF-I was similar to that after the use of Dycal. In conclusion, microscopic control of membrane sealing in situ gives valid information on the more subtle pulp effects of growth factors. The observations suggest that pulp-capping of rat molars by means of rhIGF-I enhances reparative dentinogenesis in comparison with vehicle controls. PMID- 12640755 TI - Use of electrosurgery in the perioperative setting. AB - The concept of using heat to stop bleeding goes back hundreds of years. Researchers have constructed a variety of devices that used electricity as a means to heat tissue and control bleeding. Electrosurgery became widely used in the late 1920s because of the urgent need to safely control bleeding in operative procedures. This article will cover basics of electricity and electrosurgery, clinical applications, technologies, and nursing care during electrosurgery. PMID- 12640756 TI - Protect your practice: informed consent. PMID- 12640757 TI - AORN position statements and resolution adopted. PMID- 12640758 TI - To brush or not to brush...? PMID- 12640759 TI - Glossary of legal terms. PMID- 12640760 TI - Try it, you'll like it. PMID- 12640761 TI - Sexual healing. PMID- 12640762 TI - Osteoporosis and implant prostheses. PMID- 12640763 TI - More questions and answers about the HIPAA Privacy Rule. PMID- 12640764 TI - Why the concern over mercury in wastewater? PMID- 12640766 TI - Caring for families of chronically ill kids. PMID- 12640767 TI - Are you assessing for melanoma? PMID- 12640768 TI - Clinical research: it's rewarding work. PMID- 12640769 TI - A quick guide to verbal reports. PMID- 12640770 TI - A new generation of contraceptives. PMID- 12640771 TI - Ready? HIPAA's here. PMID- 12640772 TI - Ethics, where art thou? PMID- 12640773 TI - Oral piercing: the hole story. AB - Oral and perioral jewelry is increasingly being viewed as an acceptable fashion statement in our society. Accordingly, dental professionals are being presented with questions by their patients as to the risks associated with this form of body piercing. This article reviews recommendations for individuals with, or contemplating, an oral piercing, common locations of piercings, risks associated with the piercing procedure, types of jewelry used, and post-piercing home care instructions. Current trends in the piercing industry will be discussed, and the recommendation made that dental professionals advocate for improved safety standards at body modification studios. PMID- 12640774 TI - Good people everywhere. Interview. PMID- 12640775 TI - Creating a supportive environment. An update from the Minnesota Dental Association's Committee on Environment, Wellness and Safety. PMID- 12640776 TI - "You are here"/2003: education, research, and service at the University of Minnesota School of Dentistry. Interview. PMID- 12640777 TI - Common sense, common ground, and a common language: the Health Insurance Portability and Accountability Act privacy rule. PMID- 12640778 TI - What's a dentist to do? PMID- 12640779 TI - Tobacco use patterns and attitudes among teens being seen for routine primary care. AB - PURPOSE: To describe the tobacco-related attitudes, behaviors, and needs of smoking and nonsmoking teens being seen for routine pediatric care and to identify predictors of tobacco use. DESIGN: Cross-sectional survey of adolescent primary care patients who completed self-administered questionnaires in medical office waiting rooms while waiting for routine care visits. SETTING: A group practice HMO in the Pacific Northwest. SUBJECTS: A sample of 2526 teenagers, ages 14 to 17, who consented to receive health promotion interventions as a part of a randomized trial in seven pediatric and family practice offices. MEASURES: A 38 item questionnaire assessed tobacco use history, attitudes, quit attempts, and stage of acquisition or cessation along with gender, age, race/ethnicity, body mass index, educational plans, frequency of exercise, attempts to lose weight, and depressed mood. RESULTS: Sixty-seven percent of teens approached (2526 of 3747) consented to complete a questionnaire and receive tobacco- or diet-related interventions as a part of their medical visit. About 23% of teen patients reported smoking at least one cigarette in the last month, although only 14% described themselves as current "smokers." Most current smokers (84%) smoked at least 20 days in the last month. Logistic regression predictors of smoking included older age, Native American ethnicity, lower educational aspirations, lower body mass index, smoking among half or more friends, smokers at home, and a positive depression screen. Among ever-regular smokers, most were in the action (28%), preparation (21%), or contemplation (22%) readiness to quit smoking stages, and 77% of current smokers had made one or more serious quit attempts in the last year. CONCLUSIONS: Most teens in these medical facilities consented to receive tobacco and diet interventions, and most self-described current smokers were contemplating or preparing to quit. Medical visits provide attractive opportunities for tobacco intervention, but messages should be tailored based on the patient's tobacco status and stage of acquisition or cessation. PMID- 12640780 TI - Smoking attitudes and practices among low-income African-Americans: qualitative assessment of contributing factors. AB - PURPOSE: Studies have shown that African-American adolescents are less likely to smoke cigarettes than white youth. National data suggest that this pattern changes in late adolescence and early adulthood. Specifically, African-American adults have a relatively high smoking prevalence rate when compared with other racial/ethnic groups. The purpose of this study was to qualitatively examine the sociocultural factors associated with smoking attitudes and practices among low income African-American young adults. DESIGN: Cross-sectional qualitative study. SETTINGS: High schools, 2-year colleges, housing developments, and trade schools in New Orleans, Louisiana, and Memphis, Tennessee. SUBJECTS: One hundred eighteen low-income African Americans between 18 and 35 years of age (65 men and 53 women). MEASURE/PROCEDURE: Fourteen focus groups were conducted with the target population. Nonmonetary incentives were provided for each participant in the 1 hour sessions. The majority of focus group moderators were African-American females trained in focus group moderation. Participants were recruited through flyers and project liaisons at each field location. RESULTS: Themes elicited from the focus groups were classified according to the PEN-3 model, and they included: lighting cigarettes for parents as a first experience with cigarettes, perceived stress relief benefits of smoking, use of cigarettes to extend the sensation of marijuana, and protective factors against smoking such as respect for parental rules. CONCLUSION: The results indicate that there are specific contextual and familial factors that can contribute to smoking initiation, maintenance, and cessation among low-income African-American young adults. Limitations of this study include the exploratory nature of focus groups and the relatively small sample size. Further studies are necessary to quantitatively examine the role of these factors on smoking patterns in this population. PMID- 12640781 TI - Environmental and policy factors associated with overweight among adults in Missouri. AB - PURPOSE: To measure the association between environmental and policy factors (i.e., community perceptions, community infrastructure, and worksite infrastructure) and being overweight. DESIGN: Cross-sectional data from the Missouri Cardiovascular Disease Survey, a one-time random-digit-dialed telephone interview of noninstitutionalized adults. SETTING: Missouri, 1999 to 2000. SUBJECTS: The response rate was 69.6%. A total of 2821 adults completed the interview. The sample was weighted to represent the population of Missouri: 52% female, 71% white, and 59% overweight. MEASURES: The survey comprised 92 closed ended multiple-choice items. Overweight was defined as a body mass index greater than or equal to 25 kg/m2 on the basis of self-reported height and weight. RESULTS: After adjustment for demographic and behavioral factors, environmental variables associated with being overweight included negative (i.e., unsafe and unpleasant) community perceptions (adjusted odds ratio [OR] = 1.6; 95% confidence interval [CI] = 1.1, 2.3) and the absence of outdoor exercise facilities (OR = 1.2; CI = 1.0, 1.5). Worksite policy variables were not related to the outcome; however, negative community perceptions (OR = 2.8; CI = 1.5, 5.2) and the absence of sidewalks and shoulders (OR = 1.7; CI = 1.3, 2.4) were associated with overweight among employed persons. CONCLUSIONS: An ecological perspective, focusing on the physical and social environment, was adopted to address the subject of overweight and obesity. Results of this study show that negative perceptions of the physical environment and the absence of enabling infrastructure are modestly associated with overweight, in comparison with other known risk factors. Environmental and policy interventions that promote healthier lifestyles by encouraging physical activity and healthy eating may have an effect on reducing the prevalence of overweight and obesity. PMID- 12640782 TI - Relationships of alcohol use, stress, avoidance coping, and other factors with mental health in a highly educated workforce. AB - PURPOSE: The relationships of drinking, stress, life satisfaction, coping style, and antidepressant use to mental health were examined in a highly educated workforce. DESIGN: This study used a one-time mail-out, mail-back cross-sectional survey design to examine the relationships of mental health with three kinds of stress (life events, work stress, home stress); two kinds of life satisfaction (work and home); use of avoidance coping; and antidepressant use. SETTING: This study was conducted at a large worksite in northern California in which the workforce was comprised of predominantly highly educated employees. SUBJECTS: Questionnaires were mailed to a random sample of 10% of 8567 employees, and 504 were completed and returned by participants (59%). Complete data were provided by 460 participants (53%). MEASURES: Respondents completed the Mental Health Index, the Alcohol Use Disorders Identification Test (AUDIT), and measures of coping style, work and home stress and satisfaction, stressful life events, and antidepressant use. RESULTS: Mean Mental Health Index scores were at the 32nd percentile of the U.S. population-based norms, with low percentile values associated with worse mental health. Using multiple regression analysis, the factors examined in this study were significantly related to Mental Health Index scores as the dependent variable [F(16, 443) = 27.41, p < .001, adjusted overall R2 = .48]. Poor mental health scores were significantly related to the following: age (p < .05); screening positively for current harmful or hazardous drinking (p < .05); having high levels of stress at work (p < .05) or home (p < .01); experiencing dissatisfaction with work (p < .001) or home life (p = .01); engaging in avoidance coping (p < .001); and using antidepressants (p < .001). Employees currently using antidepressants had significantly more outpatient medical and mental health visits, indicating higher health costs. Furthermore, mental health status was also significantly related to the interactions between several pairs of these variables: education and gender, age and job stress, home satisfaction and work stress, home satisfaction and avoidance coping, and home satisfaction and use of antidepressants. CONCLUSION: Mental health status was poorer on average in a highly educated workforce compared with general U.S. norms. Most of the factors that were found to be associated with poorer mental health were ones that are potentially modifiable, such as experiencing more stress and less satisfaction in work and home life and engaging in current hazardous or harmful drinking. The findings that mental health is worse among individual employees who exhibit combinations of these factors suggest that we need to better understand possible effects of these factors in the context of one another. As interpretation of these results may be limited by the single worksite that participated in this study, future research should re-examine these relationships in other worksites varying from this one in geography and demographic characteristics. PMID- 12640783 TI - Depression, functional status, treatment for psychiatric problems, and the health related practices of elderly HMO members. AB - PURPOSE: To study relationships between depression, functional limitations, psychiatric treatment, and the health-related practices of elderly individuals. DESIGN: Cross-sectional, observational study based on survey data (response rate = 90%) analyzed using multiple linear and logistic regression models. SETTING: Kaiser Permanente Northwest, a large nonprofit HMO. SUBJECTS: Medicare members (4615) aged 65-103 years who responded to a health status survey. MEASURES: Depression, health and functional status, frequency of alcohol consumption, smoking status, advance medical directive completion, influenza or pneumonia vaccinations, mammograms, recent psychiatric treatment, and attempts to lose weight or increase exercise. RESULTS: Self-reported depression was related to a greater risk of smoking (odds ratio [OR] = 1.54). Health-related functional limitations were related to fewer attempts to change health behaviors and less preventive service use (significant ORs ranged from 0.84 to 0.94). Minority group membership and Supplemental Security Income were negatively associated with better health practices and with completing advance medical directives (ORs 0.41 and 0.79, respectively). Falls in the prior year, more prescription medications, and psychiatric treatment were positively associated with better health practices (significant ORs ranged from 1.12 to 1.88). CONCLUSIONS: Depression and smoking are associated among elderly individuals, as are psychiatric treatment and attempts to lose weight. Those with functional limitations, minority status, or Supplemental Security Income use fewer preventive services; the latter two groups are less likely to complete advance medical directives. Older adults with these characteristics might need assistance in improving health-related practices. PMID- 12640784 TI - Nurses can make sure it never happens again. PMID- 12640785 TI - Combating stress. PMID- 12640786 TI - Managing toxic shock syndrome. AB - A recent case review showed that nurses play an important role in diagnosing toxic shock syndrome (TSS). It is a rare, and potentially fatal condition, which may develop in individuals of all ages. The average A&E nurse will probably only encounter a single case. However, front-line nurses will usually be the first to meet the patient with this condition. Some knowledge of TSS is important because early diagnosis and treatment may save lives. PMID- 12640787 TI - What you need to know about ... gallstones. PMID- 12640788 TI - Bed baths. PMID- 12640789 TI - Management and treatment of patients with fractured ribs. AB - Rib fractures are a common injury to the chest. The mainstay of treatment is adequate analgesia coupled with meticulous respiratory care in order to prevent complications. Treatment can be as an outpatient, but patients with more severe fractures may be admitted to hospital for oxygen therapy, ventilation, aggressive analgesic techniques and physiotherapy. Significant morbidity and mortality rates have been reported with rib fractures. To avoid complications, accurate diagnosis, adequate analgesia and effective physiotherapy are essential. PMID- 12640790 TI - Inpatient experiences of ward rounds in acute psychiatric settings. AB - This study explores inpatients' experiences of ward rounds. Eight patients on an acute admissions ward were interviewed using a semi-structured interview. Data was analysed using content analysis. The findings showed satisfaction with the ward-round process, but participants identified issues that could be improved upon, with some expressing feelings of fear and intimidation. A number of participants referred to decisions about their treatment being made before attendance and no participant expressed feeling part of the decision-making process. Despite study limitations it was concluded that some interesting themes were identified, which could be usefully explored in future studies. PMID- 12640791 TI - 'The most exciting job I ever had'. PMID- 12640792 TI - Clot busters. PMID- 12640793 TI - The challenge of wound exudate. AB - The past two decades have seen many advances in tissue viability. Our knowledge of the processes involved in wound healing has increased vastly, and as a result many products and techniques have been developed to speed it up. However, patients with wounds, and the professionals who care for them, still experience a number of challenges. PMID- 12640794 TI - Telemedicine: a new model of care. AB - An estimated one per cent of the population will have a leg ulcer and prevalence increases markedly with age (Angle and Bergan, 1997). At any one time it is estimated that 100,000 patients have open leg ulcers that require treatment. This treatment is mainly repeat dressings over extended periods of time, carried out by community nurses. This is estimated to cost 600m Pounds each year in the UK (Logan, 1997). Improving healing time for leg ulcers and reducing recurrence leads to an improved quality of life for patients and their carers in addition to major cost savings for the NHS. PMID- 12640795 TI - Managing wound exudate. AB - Wound exudate is produced as a normal part of the healing process. During the inflammatory response blood vessel walls dilate and become more porous allowing leakage of protein-rich fluid into the wounded area (White, 2000). Managing exudate and maintaining a wound environment that is moist but not wet is a constant challenge. PMID- 12640796 TI - Topical negative pressure therapy. AB - Topical negative pressure therapy (TNP) has been available for clinical use in wound management since the late 1990s. It has been shown to be of particular benefit for the treatment of chronic nonhealing wounds and has also recently been positively linked to wound bed preparation. PMID- 12640798 TI - Pressure ulcers and litigation. AB - In the past, relatives and patients appeared to accept that pressure ulcers were an inevitable result of chronic conditions and reduced mobility. Now they are viewed as evidence of a failure to provide a reasonable standard of care and action can be brought against those responsible, for compensation. PMID- 12640797 TI - Wound swabbing technique. AB - Many nurses perform clinical procedures that are based on ritual, without understanding the rationale behind them (Parker, 1999). Health policy in the UK emphasises the need for evidence-based care and requires practitioners to ensure that clinical practice is founded on scientifically-derived findings rather than intuition and ritual. PMID- 12640799 TI - A holistic approach to wound care. AB - Ms Black is a 60-year-old widow with no children, who has severe rheumatoid arthritis and uses a wheelchair. She has pulmonary disease and requires continuous oxygen therapy. For the past two years her 90-year-old mother, who has dementia, has lived with her. PMID- 12640800 TI - The management of skin tears. AB - During the ageing process the layers of the skin start to atrophy; the epidermis becomes thin and fragile, and dermal thickness decreases by 20 per cent (White et al, 1994). This makes skin tears a common problem among older people. PMID- 12640801 TI - Caring for the health of those who care. PMID- 12640802 TI - General prescribing principles. The prescription. AB - Remember ... Prescribe within your qualifications. Write clear, legible and complete prescription. You are responsible for your prescription. Good record keeping is essential. Keep prescription pads secure. PMID- 12640803 TI - Growing pains. PMID- 12640804 TI - The lost children. PMID- 12640805 TI - Work until you drop. PMID- 12640806 TI - Straight talking. PMID- 12640807 TI - Patient advocacy: a concept analysis. AB - AIM: To clarify the ill-defined concept of patient advocacy and develop a model. METHOD: An eclectic concept analysis was used in the study. RESULTS: Results of the analysis reveal that advocacy has three essential attributes: valuing, apprising and interceding. Antecedents to advocacy include a vulnerable population and a nurse willing to take on the responsibility for advocacy. The consequences of acting as a patient advocate can be potentially negative or positive for patient and nurse. On their own, the attributes are one of a number of helping strategies. CONCLUSION: Advocacy is a contemporary nursing issue comprising three essential attributes. Individually, each of the attributes is a helping strategy used in nursing. Only when all three attributes are present can advocacy be said to be realised. PMID- 12640808 TI - Post-exposure rabies vaccination. AB - Bites from dogs, cats, bats and monkeys are the most common source of rabies infection in humans. Fatal cases of rabies are relatively rare in the UK, because of ready access to post-exposure immunisation and wound treatment (DoH 2000). However, some people present to general practice seeking advice following exposure to an animal abroad. PMID- 12640809 TI - The management of prostate cancer. AB - Prostate cancer is one of the most common malignancies in westernised countries. An alarming fact is that the incidence of prostate cancer is showing an upward trend. The author discusses the anatomy and physiology, epidemiology, aetiology and management of this disease. PMID- 12640810 TI - Growing need. PMID- 12640812 TI - Reducing malpractice exposure. PMID- 12640811 TI - Incident-to billing not worth the professional price. PMID- 12640813 TI - Shared services billing. PMID- 12640814 TI - Controlling blood sugar with nutrients & botanicals. PMID- 12640815 TI - Dry at night. Treating nocturnal enuresis. PMID- 12640816 TI - Patient information. Developmental milestones for infants. PMID- 12640817 TI - Coach and activist. The NP's role in preventing youth violence. PMID- 12640818 TI - Improving child and adolescent mental health. An evidence-based approach. PMID- 12640819 TI - Family support for children with ADHD. An essential part of daily management. PMID- 12640820 TI - Too much energy for rest. Sleep problems in children with ADHD. PMID- 12640821 TI - Acne vulgaris. Current treatment approaches. PMID- 12640822 TI - Vaccine information statements. Are federal guidelines being followed? PMID- 12640823 TI - Needle stick injuries. Providers bear ultimate responsibility--and consequence. PMID- 12640824 TI - Premenstrual symptoms. Today's theories and treatments. PMID- 12640825 TI - Acupuncture. Essential information for nurse practitioners. PMID- 12640826 TI - When should you suspect endocarditis? PMID- 12640827 TI - Coronary heart disease in South Asians. PMID- 12640828 TI - A case of breathlessness. PMID- 12640829 TI - Diagnosing chronic stable angina. PMID- 12640830 TI - Cardiac rehabilitation and primary care. PMID- 12640831 TI - Inguino-scrotal problems in children. PMID- 12640832 TI - Managing sleep disorders. PMID- 12640833 TI - Passing the simulated surgery module. PMID- 12640834 TI - Stay visible! PMID- 12640835 TI - The influence of DNR orders on patient care in adult ICUs: a review of the evidence. AB - BACKGROUND: Although do-not-resuscitate (DNR) orders have been used in health care for many years, there is controversy about their use and the implications for patient care. A common perception in the intensive care unit (ICU) setting is that life-sustaining interventions such as dialysis or mechanical ventilation may no longer be provided or should be withdrawn on patients who are assigned DNR status. In order to gain insight into the impact of DNR orders on the care provided to adult patients in the ICU, a review of the research conducted on this topic within the last decade was undertaken. OBJECTIVE: To provide an integrated review of recent research examining the effects of DNR orders on the care of adult ICU patients. METHODS: Two large databases, CINAHL and MEDLINE, were used for the search as well as the reference lists from relevant research articles. The search terms used were: DNR orders in adult ICUs, resuscitation orders, critical care, and nursing and/or medical care provided to these patients. Studies were not limited to any one specific research design. The search was restricted to research conducted over the last decade. RESULTS: Five studies were found that met the search criteria. CONCLUSIONS: While the care that critically ill adult patients receive in ICUs is, in some way, influenced by DNR orders, there is no evidence that patients are abandoned as a result. Given the inconsistency in the findings of the five studies and certain weaknesses in methodology, it is not possible to identify a direct impact of DNR orders on patients care at this time. Further investigation is needed. PMID- 12640836 TI - The CNA national nursing forum. PMID- 12640837 TI - Pediatric critical care examination in 2003. PMID- 12640838 TI - A number of ICUs have received boards from the CACCN national office that they have mounted on the wall in their unit. What is the purpose of these boards and how does an ICU go about acquiring one? PMID- 12640839 TI - A review of neuromuscular blockade in the critically ill patient. PMID- 12640840 TI - ACLS 2000: overview of changes to the guidelines. PMID- 12640842 TI - Wrapping up.... PMID- 12640841 TI - Aprotinin: antifibrinolytic and anti-inflammatory mechanisms of action in cardiac surgery with cardiopulmonary bypass. AB - Cardiopulmonary bypass results in activation of the coagulation, fibrinolytic, inflammatory, and complement cascades. These activated cascades result in a decrease in the number of circulating coagulation factors, hyperfibrinolysis, thrombocytopenia, platelet defects, coagulopathies, and an acute inflammatory response. Patients experiencing cardiac surgery with cardiopulmonary bypass are at risk for many potential problems. The use of aprotinin, an antifibrinolytic agent, has multiple effects that tend to reduce hematological defects and blunts the inflammatory response that is associated with cardiac surgery. The pathophysiological consequences of cellular activation associated with cardiopulmonary bypass, basic mechanisms of action of aprotinin, pharmacodynamic and pharmacokinetic properties, dosing, adverse reactions, and cost/benefit ratio are discussed in this article. Critical care nurses need to know about aprotinin to understand its role in reducing blood loss and transfusions during and after cardiac surgery. PMID- 12640843 TI - What action, if any, did the board of directors take regarding Bill 85 in Ontario: "the Patient Restraints Minimization Act"? PMID- 12640844 TI - Refining your research question. PMID- 12640845 TI - New drug approvals in 2001. PMID- 12640846 TI - Prescribing for patients who aren't there. PMID- 12640847 TI - Refining the art of porcelain laminate veneers. AB - As with any other operative or restorative technique, the long-term results, both esthetic and functional, depend directly on [figure: see text] the effort made by the dentist. Attention to detail, allowing enough time, and a steady hand are necessary to obtain the desired result. A few operative tips have been provided to aid the dentist in achieving these sometimes-elusive maximum results. Proper diagnosis, canine-guided occlusion, and attention to detail will help you give your patients that dazzling new smile! PMID- 12640848 TI - New caries detection technologies and modern caries management: merging the strategies. AB - Literature over the past decade has suggested that dental caries be treated as a curable and preventable infectious disease. The emphasis is placed on early detection and intervention, including chemical therapies and minimally invasive strategies. There are now new detection technologies commercially available to help detect carious lesions early; however, what seems to be lacking is a clear strategy on how to merge these new technologies in a way consistent with today's changing paradigm of caries management. There is a real danger that these early detection technologies may be used to unnecessarily and overaggressively "drill and fill" using Black's "extension for prevention" preparations. This paper will review the literature and key strategies for caries management and early detection and suggest one way (not necessarily the only way) in which they may be used in harmony. Two relatively new detection technologies (DIAGNOdent and DIFOTI) will be reviewed. PMID- 12640850 TI - In vitro testing of xylitol as an anticariogenic agent. AB - A number of studies involving xylitol chewing gum have demonstrated that xylitol is both noncariogenic and anticariogenic. The ability of xylitol to act as an anticariogenic agent most likely is due to its ability to be transported into caries-causing oral bacteria and inhibiting fermentation either by depleting the cell of high-energy phosphate or by poisoning the glycolytic system. In vitro tests were conducted to determine the concentration of xylitol required to inhibit the growth of three strains of oral streptococcus (S. mutans, S. salivarius, and S. sanguis). All three strains were inhibited significantly at xylitol concentrations of 12.5% and higher; however, only S. mutans was inhibited significantly at a xylitol concentration of 1.56%. PMID- 12640849 TI - A paradigm shift in the treatment of caries. AB - This paper presents a shift in the paradigm for treating dental caries from a surgical to a medical approach. Two key points are essential to understanding caries. First, caries is a bacterial infection caused by specific bacteria. Second, before cavitation, caries is a reversible multifactoral process. Two features, caries activity and caries risk, provide a diagnosis and prognosis leading to a treatment plan. Patients receive treatment using a medical model consisting of four steps: bacterial control; reduction of risk levels for at-risk patients; reversal of active sites by remineralization; and follow-up and maintenance. PMID- 12640851 TI - Detection of occlusal carious lesions. AB - The purpose of this study was to determine the sensitivity and the specificity of detecting the presence and extent of occlusal carious lesions using traditional examination techniques performed by three experienced restorative dentists and a laser fluorescence-measuring device (DIAGNOdent) on 100 extracted third maxillary and mandibular molars. The extracted teeth were selected to ensure presence of questionable occlusal caries and intact occlusal surfaces. The results revealed that the DIAGNOdent demonstrated a higher sensitivity (94%) compared to the clinicians' consensus (82%) but a lower specificity. This study also indicated that higher values of the DIAGNOdent do not appear to correlate well with the depth of the carious lesion in the dentin (r = 0.3809) as measured by a caries detecting dye. PMID- 12640852 TI - Degree of fluorosis in areas of South Africa with differing levels of fluoride in drinking water. AB - The purpose of this study was to study the relationship between degree of fluorosis and varying fluoride concentrations in the drinking water in a relatively dry region in South Africa. A strong positive association was found between drinking water fluoride levels and fluorosis but only up to a level of approximately 2.0 ppm for areas with a high average daily temperature (approximately 79 degrees F). The results indicate that the optimum drinking water fluoride concentration for a region with a high average daily temperature should be less than 1.00 ppm. PMID- 12640853 TI - Dental needs of the elderly in the 21st century. AB - The average age of the world's population is increasing rapidly. The "graying of America" presents new opportunities and new challenges for improving the oral health of the elderly, particularly those afflicted with neurocognitive impairments. The dental problems associated with these conditions include but are not limited to a decrease in oral hygiene; difficulty in controlling and retaining dentures; xerostomia, which often is drug-associated, and consequential root caries, recurrent decay, and purposeless chewing. Parkinson's disease and Alzheimer's disease are the most prevalent type of progressive neurocognitive impairing illnesses, affecting millions of elderly Americans. As the adult population increases, a greater number of patients with these diagnoses will require dental care. Dental providers need to be aware of the special problems associated with the treatment of the older healthy subject and the neurocognitively impaired patient. PMID- 12640854 TI - Glass ionomer endodontic sealers--a literature review. AB - The properties of endodontic glass ionomer sealers are summarized and compared to properties of Grossman's ideal sealer and a typical ZOE sealer. A review of the literature regarding glass ionomer and other endodontic sealers also is included. Endodontic glass ionomer sealer appears to be an alternative to conventional ZOE sealers in canals that are adequately isolated, instrumented, and dried. PMID- 12640856 TI - Red, bumpy palate. Inflammatory papillary hyperplasia. PMID- 12640855 TI - Anesthetic efficacy of different volumes of lidocaine with epinephrine for inferior alveolar nerve blocks. AB - Volumes of 1.8 mL and 3.6 mL of 2% lidocaine with 1:100,000 epinephrine were compared in inferior alveolar nerve blocks. The first molars, first premolars, and lateral incisors of 462 subjects were pulp tested for 55-60 minutes following administration of inferior alveolar nerve blocks. Results showed no significant difference between the two volumes of anesthetic in achieving successful pulpal anesthesia. PMID- 12640857 TI - Rough white patch of gingiva. Verruciform xanthoma. PMID- 12640858 TI - A call for nursing research in assisted living facilities. PMID- 12640859 TI - Conducting research related to treatment of Alzheimer's disease. Ethical issues. AB - Researchers are obligated to protect the rights of study participants. Protecting the rights of patients with Alzheimer's disease (AD) is particularly complicated because of the special needs of this patient population, and the characteristics of developing treatments and technologies. Respecting autonomy and the right to self-determination are complicated by difficulties associated with assuring competence, understanding, and voluntariness in the informed consent process. Protecting patients with AD from harm may be complicated because new treatments have subtle side effects that may be difficult to detect in patients experiencing communication difficulties. Harm to patients with AD also may occur from withholding proven treatments in placebo-controlled trials, and in the use of genetic testing. Issues of justice in the allocation of research dollars and the ability of patients with AD to participate in research are also discussed. By recognizing potential pitfalls, researchers involved in testing new treatments for patients with AD can take proper steps to assure ethical treatment of study participants. PMID- 12640860 TI - Health and safety risk at a skilled nursing facility. Nursing assistants' perceptions. AB - Little is known about the perceptions nursing assistants certified (NAC) have related to their occupational health and safety risks and how these perceptions influence the safety precautions taken. The purpose of this study was to describe the perceptions of the occupational health and safety risks perceived by NAC, along with determining the protective mechanisms they used to prevent injury and illness. Ten NAC employed by a skilled nursing facility were interviewed using a semi-structured interview guide. Numerous hazards and protective mechanisms were identified and these were categorized and environmental, physical or cognitive in nature. Based on the NAC perceptions of their occupational hazards, there were a number of areas in which they were ignorant to concepts the administration believed to be readily available. While the facility has a number of health promotion and safety programs already in place, these alone seem inadequate to meet the perceived needs of the NAC. Strategies are needed in three specific areas to address the knowledge deficits and enhance the overall effectiveness of improving the health and safety of the NAC--communication, fostering teamwork, and educational and training opportunities. PMID- 12640861 TI - Mahler's developmental theory. Training the nurse to treat older adults with borderline personality disorder. PMID- 12640862 TI - Family caregiver assessment. Essential for effective home health care. AB - Home health care nurses do not routinely assess the health of family caregivers despite their essential contribution to the client's care. In this study, home care nurses collected data on 51 older family caregivers from their caseloads to assess their health. The average age of the caregivers was 75.1 years (SD = 6.09). Most (66.7%) were women and were the home health client's spouse (82.4%). These individuals had been caregivers for up to 20 years, and reported spending an average of 13.3 hours per day in this role (SD = 9.15). Approximately half (n = 25) of the caregivers reported poor or fair health, with 33.3% (17) reporting a decline in their health over the previous 6 months. Unmet health needs included the need for blood pressure monitoring, mammograms, PAP smears, and prostate examinations. Referrals to other health care providers or community agencies were required by 78.4% of the caregivers, 78.4% required health teaching, and 23.5% required home health services themselves. Those requiring home health care were more likely to be on more medications and classify their health as fair or poor. These findings confirm those of a pilot study conducted on 51 other family caregivers. Nurses serving the geriatric population need to conduct systematic family caregiver assessments to identify caregiver health needs that could impair their caregiving ability. In particular, for home health care to be effective, nurses must conduct caregiver assessments. PMID- 12640863 TI - Group diabetes patient education. A model for use in a continuing care retirement community. PMID- 12640864 TI - Comparison of family caregivers. Stroke survivors vs. person with Alzheimer's disease. AB - The purpose of this study was to compare family caregivers of person with dementia and caregivers of stroke survivors with similar levels of caregiving responsibilities. The design was descriptive and included characteristics of care recipients, caregivers, and care situations. Caregivers of individuals with dementia reported care recipients were more impaired with independent activities of daily living and memory and behavior problems. There were no differences in caregiver depression and fatigue. Even with the benefit of respite care, a substantial number of caregivers had depression scores above the level indicating possible clinical depression. Consultation by advanced practice psychiatric nurses for caregivers and care recipients may be beneficial in detecting depression and making recommendations for appropriate treatment. PMID- 12640865 TI - Intravenous infusion--the case for keeping vitamin C in the emergency drug cupboard. PMID- 12640866 TI - Bone densitometry--role of quantitative CT. PMID- 12640867 TI - Bone densitometry--role of quantitative CT. PMID- 12640868 TI - Legal termination of pregnancy. PMID- 12640869 TI - The area needs certificate. PMID- 12640870 TI - Nuclear medicine in South Africa. PMID- 12640871 TI - Ethical standards. PMID- 12640872 TI - The Employment Equity Act--putting the record straight. PMID- 12640873 TI - Tackling the crowded jail health threat. PMID- 12640874 TI - Birth deformities--a heavy burden. PMID- 12640875 TI - Sleep disorder adds to disease burden. PMID- 12640876 TI - Summarised highlights of the latest amendments to the regulations of the Medical Schemes Act: Part II. PMID- 12640877 TI - Models of practice management. PMID- 12640878 TI - Egg allergy and MMR vaccination. PMID- 12640879 TI - What do South African psychiatrists and GPs think, feel and know about evidence based mental health care? PMID- 12640880 TI - Eczema and foods. PMID- 12640881 TI - South Africa's role in medical training in Malawi. PMID- 12640882 TI - A frightening situation? Youth in the USA today. PMID- 12640883 TI - Permeability of intestinal mucosa to crystalline and tabletted isoniazid (INH). PMID- 12640884 TI - Clinically silent renal tumour producing erythropoietin. PMID- 12640885 TI - Amoebic liver abscess--results of a conservative management policy. AB - OBJECTIVE: To evaluate the safety and efficacy of conservative management of amoebic liver abscesses. DESIGN: A prospective study carried out over a 1-year period. SETTING: Inpatients and outpatients in a tertiary referral institution. SUBJECTS: Amoebic liver abscess was diagnosed on clinical, ultrasonographic, and serological features. All patients were treated with metronidazole. The indication for ultrasound-guided aspiration of the abscess was failure to improve clinically within 48-72 hours. MAIN OUTCOME MEASURES: Clinical improvement, clinical deterioration and failure of clinical improvement (persistent pain). RESULTS: In total 178 patients (male-to-female ratio 5:1) with 203 abscesses were treated during this period. Of these, 23 patients required percutaneous aspiration and 150 patients were managed without intervention and clinically resolved spontaneously. Abscesses requiring aspiration tended to be larger than those managed without aspiration (10.7 cm v. 8.2 cm) (p = 0.003). There were no complications following aspiration. Mean hospital stay was longer (12.3 days) for patients who underwent aspiration compared with those who did not (6.7 days) (p = 0.031). Only 5 patients presented with ruptured abscesses, 1 cutaneously and 4 intraperitoneally, with the only death in this latter category. CONCLUSION: Conservative medical management of amoebic liver abscess is safe. Percutaneous ultrasound-guided aspiration is indicated only in patients who fail to improve clinically after 48-72 hours rather than on rigid criteria. PMID- 12640886 TI - An epidemiological perspective of substance use among high school pupils in rural KwaZulu-Natal. AB - OBJECTIVES: To investigate prevalence and the factors influencing substance use among rural high school pupils in KwaZulu-Natal in order to develop and implement intervention programmes. DESIGN: Cross-sectional study. SETTING: Twenty-eight high schools in southern KwaZulu-Natal. SUBJECTS: One thousand three hundred and eighteen grade 10 pupils. OUTCOME MEASURES: An anonymous self-reporting questionnaire was used to investigate the use of alcohol, tobacco (cigarettes), cannabis and solvents. RESULTS: Of the male scholars, 52.9% (95% confidence interval (CI): 45.4-60.3) reported ever using alcohol, 16.9% (CI: 11.5-24.0) reported using cannabis, and 13.1% (CI: 7.2-22.5) had smoked more than one cigarette daily. Among male pupils 45.5% (CI: 38.6-52.6) had inhaled benzine and 34.6% (CI: 28.0-41.8), thinners; 7.4% (CI: 3.4-15.2) had used cocaine and 4.1% (CI: 1.0-10.6), crack. Female pupils reported significantly less use of alcohol (25.5%, CI: 17.6-35.3), cannabis (2.3%, CI: 1.3-4.1), cigarettes (more than one daily) (2.0%, CI: 1.0-4.1), and inhalation of benzine (18.8%, CI: 13.8-25.2) and thinners (10.8%, CI: 7.2-16). Logistical regression indicated that the odds of smoking cigarettes increased significantly (p < 0.0005) with use of the other substances. CONCLUSION: The results of this study confirm the prevalence of multi substance use among pupils at the majority of rural high schools in this district and the need for targeted interventions to reduce/prevent this. PMID- 12640887 TI - Lessons learned from infertility investigations in the public sector. AB - OBJECTIVES: To determine the main factors causing infertility in an urban, tertiary hospital population. To establish if any such major causal factor could be used to rationalise and improve the service for infertile couples in the public sector. DESIGN: A retrospective analysis of the hospital records of 206 women who had a tubal patency test (hysterosalpingogram) performed and the results of the investigations performed in the couples with infertility. RESULTS: Of the 206 women 38 (18.5%) had normal fallopian tubes on hysterosalpingogram; 33 (16%) had unilateral obstruction and 135 (65.5%) had bilateral tubal obstruction. Of the latter group 81 (60%) had significant hydrosalpinges. Semen analysis results in 148 partners (71.8%) demonstrated a normal count in 85 (62%), normal motility in 70 (51%) and normal morphology in only 25 (18%). Testing for ovulation (mid-luteal phase progesterone) was positive in 91 of 124 women tested (73%). Compliance, technical and logistical problems were encountered with both semen analysis and mid-luteal phase progesterone tests. CONCLUSIONS: Infertility is a major problem in South Africa, with limited resources for investigation and treatment in the public sector. Tubal factor infertility was the most common cause of infertility demonstrated in this study. In the presence of bilateral tubal obstruction with hydrosalpinges the prognosis is so poor that unless assisted reproductive techniques are available and affordable, further infertility investigations do not seem justified. Recommendations on an approach to the infertile couple in the public sector is outlined. PMID- 12640888 TI - Cancer patterns in four districts of the Transkei region--1991-1995. AB - BACKGROUND: Oesophageal cancer (OC) is an important public health problem among the Xhosa-speaking people of the Transkei region in the Eastern Cape Province, South Africa, with incidence rates for males among the highest in the world. OBJECTIVES: To record the occurrence of cancer among men and women of all ages in four districts in the Transkei during the period 1991-1995, to identify common cancers and to compare the variations in cancer incidences in this region with incidences in Africa and the rest of the world. DESIGN: Cancer registration of cases reported from all clinics and hospitals was conducted in the four selected districts. SETTING: The districts included Centane (Kentani), Butterworth, Bizana and Lusikisiki in the Transkei region. METHODS: Active and passive methods were used to collect data, which were analysed using the Statistical Analyses Systems (SAS) package. RESULTS: The mean annual number of all cancer cases reported was 310, with age-standardised incidence rates (ASIRs, world standard) of 98.2/100,000 and 74.3/100,000 for males and females, respectively. The most frequently reported cancer was OC, with mean annual ASIRs of 76.6/100,000 and 36.5/100,000 for males and females, respectively, with a male/female ratio of 2:1. CONCLUSION: The present data confirm previous reports that OC rates in Centane have consistently remained very high, whereas time-dependent changes in the incidence of OC have occurred in Butterworth, Bizana and Lusikisiki suggesting changes in the risk determinants in these districts. PMID- 12640889 TI - The relentless spread of tuberculosis in Zambia--trends over the past 37 years (1964-2000). AB - OBJECTIVE: To review trends in the rates of tuberculosis (TB) case notifications over a 37-year period. DESIGN: A retrospective study of Ministry of Health records on TB notifications between 1 January 1964 and 31 December 2000. SETTING: Zambia, sub-Saharan Africa. METHODS: Retrospective analysis of case-notification data for TB of the Zambia Ministry of Health annual returns. OUTCOME MEASURES: Annual TB case-notification rates and trends over the past 37 years. RESULTS: TB case-notification data from 1964 to 2000 show a 12-fold increase over the past two decades, and apparent gains in controlling TB seen in the 1960s and 1970s have been reversed over the past two decades. A stable situation during the period 1964-1984 (case-notification rate remained around 100 per 100,000 population) was followed by an exponential increase since the mid-1980s. The absolute number of new TB cases increased from 8,246 in 1985 (124/100,000) to 38,863 (409/100,000) in 1996 and 52,000 (512/100,000) in 2000. Comparison of case notification rates over the past 2 decades with neighbouring countries (Zimbabwe, Malawi and Tanzania) show that Zambia has one of the highest case-notification rates in the region. CONCLUSIONS: Zambia, like many countries in Africa, is in the midst of a serious TB epidemic and there are no signs that it is abating. This increase was most likely due to the impact of the HIV/AIDS epidemic and subsequent breakdown of TB services. Concerted donor-government efforts should invest appropriately in long-term plans for TB control. PMID- 12640891 TI - [Scientific practice]. PMID- 12640890 TI - Percutaneous arthrodesis. AB - It has been generally accepted that residual cartilage and subchondral bone has to be removed in order to get bony fusion in arthrodeses. In 1998 we reported successful fusion of 11 rheumatoid ankles, all treated with percutaneous fixation only. In at least one of these ankle joint there was cartilage left. This was confirmed by arthrotomy in order to remove an osteophyte, which hindered dorsiflexion. More than 25 rheumatoid patients with functional alignment in the ankle joint have subsequently been operated on with the percutaneous technique, and so far we have had only one failure. Patients with rheumatoid arthritis are known to sometimes fuse at least their subtalar joints spontaneously, and the destructive effect of the synovitis on the cartilage could contribute to fusion when using the percutaneous technique. In a rabbit study we therefore tested the hypothesis that even a normal joint can fuse merely by percutaneous fixation. The patella was fixated to the femur with lag screw technique without removal of cartilage, and in 5 of 6 arthrodeses with stable fixation bony fusion followed. Depletion of synovial fluid seemed to be the mechanism behind cartilage disappearance. The stability of the fixation achieved at arthrodesis surgery is an important factor in determining success or failure. Dowel arthrodesis without additional fixation proved to be deleterious. A good fit of the bone surfaces appears necessary. In the ankle joint, it would be technically demanding to retain the arch-shaped geometry of the joint after resection of the cartilage. Normally the joint surfaces are resected to produce flat osteotomy surfaces that are thus easier to fit together, encouraging healing to occur. On the other hand it is considered an advantage to preserve as much subchondral bone as possible, as the strong subchondral bone plate can contribute to the stability of the arthrodesis. Ankle arthrodesis can be successfully performed in patients with rheumatoid arthritis by percutaneous screw fixation without resection of the joint surfaces. This procedure has two advantages: first, it is less surgically traumatic, second, both the arch-shaped geometry and the subchondral bone are preserved, and thus both could contribute to the postoperative stability of the construct. Intuitively, preservation of the arch-shape should increase rotational stability. The results of our experimental sawbone study indicate that the arch shape and the subchondral bone should be preserved when ankle arthrodesis is performed. The importance of this is likely to increase in weak rheumatoid bone. In a finite element study the initial stability provided by two different methods of joint preparation and different screw configurations in ankle arthrodesis, was compared. Better initial stability is predicted for ankle arthrodesis when joint contours are preserved rather than resected. Overall, inserting the two screws at a 30-degree angle with respect to the long axis of the tibia and crossing them above the fusion site improved stability for both joint preparation techniques. The question rose as to whether patients with osteoarthritis could also be operated on solely by percutaneous fixation technique. The first metatarsophalangeal joint in patients with hallux rigidus was chosen as an appropriate joint to test the percutaneous technique. In this small series we have shown that it is possible to achieve bony fusion with a percutaneous technique in an osteoarthrotic joint in humans, but failed to say anything about the fusion rate. PMID- 12640892 TI - [Money or life?]. PMID- 12640893 TI - [How much does the society want to pay?]. PMID- 12640895 TI - [Diagnosis of renal artery stenosis with Doppler sonography]. PMID- 12640894 TI - [Gaucher disease]. PMID- 12640896 TI - [The importance of adipose tissue for development of obesity and diabetes mellitus type 2]. PMID- 12640897 TI - [Isoprostanes--new markers of oxidative stress]. PMID- 12640898 TI - [Transmission of methicillin resistant Staphylococcus aureus to primary health care workers in a home environment]. PMID- 12640899 TI - [Knee injuries in the Oslo emergency ward]. PMID- 12640900 TI - [Transfusion of "old" blood from the blood banks--time for a new strategy?]. PMID- 12640901 TI - [Gynecological laparoscopy in Norway]. PMID- 12640902 TI - [Adverse drug reaction reporting and drug monitoring]. PMID- 12640903 TI - [Scientific criteria methods for estimating and evaluation of screening]. PMID- 12640904 TI - [Evaluation of new laboratory tests]. PMID- 12640905 TI - [New health technologies--from development to everyday practice]. PMID- 12640906 TI - [Physicians and research--when is the lowest point reached?]. PMID- 12640907 TI - [Health service research in American backwoods]. PMID- 12640908 TI - [Financing of enzyme treatment in Gaucher disease]. PMID- 12640909 TI - [Autopsies at Norwegian hospitals]. PMID- 12640910 TI - [Sharp condemnation, not a cold professional assessment!]. PMID- 12640911 TI - [Biopsychosocial medicine and phenomenology]. PMID- 12640912 TI - The gentle touch. PMID- 12640913 TI - Capital transfer. PMID- 12640914 TI - A quiet word. PMID- 12640915 TI - Patient choice. Something to write home about. PMID- 12640916 TI - Open space. Support system. PMID- 12640917 TI - Workforce planning. A force for good. AB - A workforce development confederation facing a shortage of 2,000 clinical staff by 2004 has initiated a programme to train assistant practitioners. The programme is aimed at drawing workers into the NHS who have some care experience but not the educational requirements for traditional professional courses. The project, which involves 14 organisations across health and social care, has attracted 216 entrants. The course takes two years full time, or three years part time. PMID- 12640918 TI - HSJ event. Catching the drift. PMID- 12640919 TI - Human resources. In the eye of the storm. PMID- 12640920 TI - New-venture profits help balance SNF shortfalls. PMID- 12640922 TI - OIG 2003 work plan ready for SNFs. PMID- 12640921 TI - Measuring quality with QMs. PMID- 12640924 TI - Worrying findings require action. PMID- 12640925 TI - All together now? PMID- 12640923 TI - When the HIPAA rule applies. PMID- 12640926 TI - Country life. PMID- 12640927 TI - Care provision in Scotland: background, policy context and research. PMID- 12640928 TI - Supporting roles within a stroke rehabilitation setting. PMID- 12640929 TI - Learning from carers' experiences: helping older people with dementia to eat and drink. AB - Eating and drinking difficulties are common in people with dementia and yet some health staff are inclined to neglect this important area of care. This article explores how, combined with the skills of nurses, the experiences of those caring for relatives with dementia can help potential problems to be avoided. PMID- 12640930 TI - Assessing fluid balance in older people: fluid replacement. PMID- 12640931 TI - Highland leader. Interview by Daniel Allen. PMID- 12640932 TI - It's not what you do--it's the way that you do it. PMID- 12640933 TI - Winning the race. PMID- 12640934 TI - Developing education and support for community nurses. Principles and practice of palliative care. PMID- 12640935 TI - Recruitment and retention in the NHS. A study in motivation. PMID- 12640936 TI - Leadership in the care home sector. PMID- 12640938 TI - Communication and managerial effectiveness. PMID- 12640939 TI - Where are we now? The status of nurse practitioner practice in statute and regulation December of 2002. PMID- 12640940 TI - Diagnosis and treatment of hypertension in children and adolescents. AB - PURPOSE: To provide nurse practitioners (NPs) with updated information regarding the etiology, diagnosis, and treatment of childhood hypertension. DATA SOURCES: Extensive review of the scientific literature regarding hypertension, including the latest NIH recommendations. CONCLUSIONS: Hypertension affects more than 350,000 American children. While the majority of hypertension in early childhood occurs from secondary causes, the incidence of essential hypertension in later childhood and adolescence is rising, raising concerns as elevated pressures in childhood "track" into adulthood. Early detection and treatment of elevated childhood pressures represent important steps in reducing long-term cardiovascular risk. IMPLICATIONS FOR PRACTICE: NPs must be able to accurately differentiate between primary and secondary hypertension in childhood. Secondary hypertension requires prompt diagnosis and treatment, and controlling primary childhood hypertension has lifelong implications. Given the familial predisposition to hypertension, it is important for adult NPs to be aware of the risks faced by children of hypertensive patients. PMID- 12640941 TI - Interstitial cystitis. AB - PURPOSE: To describe the pathophysiology, assessment, diagnosis, and management of interstitial cystitis. DATA SOURCES: Selected professional publications and presentations. CONCLUSIONS: Interstitial cystitis (IC) is a chronic and disabling condition. Symptoms include urinary frequency and urgency, pain, dyspareunia, and nocturia. It occurs primarily in women. Onset is predominately in adulthood although IC does occur in childhood. Bladder wall defects, auto-immune disorder, viral and/or bacterial infection, toxin exposure, pelvic floor dysfunction and inflammatory response are possible causes. Diagnosis is by history, physical examination, laboratory tests, and cystoscopic examination. The management of interstitial cystitis includes dietary changes, antihistamines, tricyclic antidepressants, oral and intravesicle glucosaminoglycans, hydrodistention, pain management and emotional support. IMPLICATIONS FOR PRACTICE: Nurse practitioners need to be knowledgeable about the occurrence and debilitating effect of IC. Inclusion of IC into the differential diagnoses related to urgency, frequency, and abdominal pain will help assure the timely and effective diagnosis and management of this unusual disease. PMID- 12640942 TI - Myasthenia gravis: diagnosis and treatment. AB - PURPOSE: To provide information about myasthenia gravis (MG), including pathophysiology, differential diagnosis, treatments, diagnostic work up, patient teaching, and diagnostic algorithms for ptosis and weakness related to MG. DATA: Clinical and research journal articles, medical textbooks, and a patient with MG. CONCLUSIONS: Myasthenia Gravis is a very complex disease with many different signs and symptoms, which may be quite noticeable or vague. The presenting symptoms of ptosis and/or diplopia should cause the clinician to consider a work up for MG. IMPLICATIONS FOR PRACTICE: Because of the low incidence MG (1 in 10,000) has in the U.S., a delay in diagnosis of one to two years is not uncommon. It is vital to diagnose this disease and begin treatment as soon as possible to decrease the signs and symptoms. PMID- 12640943 TI - Cognitive function assessment in individuals at risk for Alzheimer's disease. AB - PURPOSE: To describe brief assessments of functional performance and cognition to detect Alzheimer's disease (AD) and depression in older adults for the primary care provider. DATA SOURCES: Review of the literature. CONCLUSIONS: It is important for the clinician to interview both the patient and a knowledgeable informant to assess changes in the patients' functioning in daily life tasks and to administer brief screening tests to detect cognitive impairment and depression in older adults. Several suggested instruments for use in the primary care setting are included. IMPLICATIONS FOR PRACTICE: Standardized assessments of functional performance and brief cognitive tests identify individuals with clinically meaningful cognitive impairment and provide baseline measurement against which to compare future assessments. PMID- 12640944 TI - Using the nursing interventions classification to describe the work of the nurse practitioner. AB - PURPOSE: To extend the specialty use research based on the Iowa Intervention Project using the Nursing Interventions Classification (NIC) and identify and the 20 most prevalent interventions described by practicing nurse practitioners (NPs). DATA SOURCES: A descriptive survey using a mailed questionnaire was sent to 1,190 NPs with prescriptive authority in Minnesota. Results are based on 414 (37%) useable responses. CONCLUSIONS: Of the 486 NIC interventions on the questionnaire, NPs reported using an average of 120 at least once per month. The 20 most frequently selected were reported by 71%-90% of respondents as being used at least once per month. Four core interventions (documentation, telephone consultation, teaching: prescribed medication, and emotional support) were common to NPs in all specialties. IMPLICATIONS FOR PRACTICE: Common medical coding does not reflect the nursing aspects of NP practice. As NPs attempt to define what is different about NP practice, the use of a standardized nursing language such as the NIC may facilitate this process. One clarification that needs to be made with regard to the NIC in future research with NPs is the distinction between prescribing and performing the interventions. PMID- 12640946 TI - Streptococcal fasciitis with toxic shock syndrome in the pediatric patient. AB - Necrotizing fasciitis (NF) is a distinctive soft tissue infection usually caused by Group A beta-hemolytic streptococcus, often seen in association with varicella in the previously healthy child. Its fulminant course is associated with great morbidity and high case-fatality rates, especially when it occurs in conjunction with streptococcal toxic shock syndrome. It differs from simple cellulitis in clinical presentation, appearance on examination, and need for urgent surgical intervention. The patient is irritable and complains of severe pain in the involved site, which is usually the extremity. On examination, exquisite pain on palpation of the involved site is confirmed, which is out of proportion to the cutaneous findings. Shock, multiorgran failure, and death will ensue if the diagnosis is not promptly recognized. The five keys to management include fluid management, aggressive debridement of necrotic tissues, anticipation and management of multisystem organ failure, appropriate parenteral antimicrobial therapy using penicillin and clindamycin, and use of intravenous immunoglobulin (IVIG). PMID- 12640947 TI - Bicycle injuries and safety helmets in children. Review of research. AB - Bicycle injuries are the most common cause of serious head injury in children, and most of these injuries are preventable. The protective effect of bicycle helmets is well documented, but many child bicyclists do not wear them. This article summarizes the current state of research on bicycle injuries and helmet use and examines the effectiveness of legislation and injury-prevention strategies. Current studies indicate that children who wear helmets experience fewer head injuries and decreased severity of injury. Community-wide helmet promotion campaigns combined with legislation are most successful in increasing helmet use and decreasing injury. Nurses can participate both at the institutional level and in community advocacy groups to promote bicycle safety for children. PMID- 12640949 TI - Child and adolescent violence. AB - Although the juvenile violent crime rate has decreased steadily during the past 5 years, the problem of violence and violence-related behaviors in the lives of our children and adolescents remains. The incidence of violent victimization against children and violence and violence-related behavior by today's youth is related to a variety of factors. Exposure to violence in the home, school, community, or video games and other entertainment significantly influences aggressive behaviors among children and adolescents. Other childhood violence predictors include alcohol and drug use, gender, and low self-esteem. The childhood violence risk indicators have implications for child and adolescent violence prevention and intervention programs. Nurses who recognize dangerous and potentially dangerous behavior in children and adolescents are better able to provide violence prevention and intervention services and referrals to children at risk or in danger. Because orthopaedic nurses often see adolescents who have already sustained injury from violence, identification of those at risk is particularly important. PMID- 12640948 TI - Gout: not just for the rich and famous! Everyman's disease. AB - Gout is a painful form of arthritis that affects more than a million Americans annually. Using a case study, a discussion of the clinical onset, signs and symptoms, diagnosis, and treatment of acute and chronic gout is presented. The incidence and etiology of gout is presented, along with current diagnostic criteria and the most common differential diagnoses to consider. The current trends in diagnosing and treating the acute and chronic forms of gout are also addressed as well as some of the controversies and concerns about treatment options. The treatment plan and follow-up for the patient in the case presentation conclude the discussion. PMID- 12640950 TI - Use of evidence for prevention of work-related musculoskeletal injuries. AB - Work-related injuries among nurses are a complex and costly problem. To thoughtfully approach this issue, one medical center developed a set of injury reduction strategies and related tools based on a review of research literature and locally obtained evidence. This article describes the evidence-based approach to prevention of work-related musculoskeletal injuries. It also covers research based observations about prevention strategies; provides a description of selected interventions, such as a patient handling guide; and includes sample tools that enable collection of meaningful data about local patient handling practices. PMID- 12640951 TI - Patient empowerment after total hip and knee replacement. AB - PURPOSE: This research was designed to gain an enhanced understanding of empowerment within in-home care relationships after hospital discharge of elderly patients who had undergone total hip or total knee replacement. METHOD: An interpretive phenomenology method was used. SAMPLE: Nine participants were interviewed on audiotape, guided by a semistructured interview guide. Five themes emerged centering on a strong desire to maintain independence. Overall, participants experienced disempowered relationships with professional in-home care providers and a more equitable empowered relationship with nonprofessional care providers. CONCLUSION: Disempowered relationships with professional personnel were not identified as a dissatisfier to most participants. Rather, patients expressed deference to the traditional expert model of healthcare. Further investigation is needed to understand the effect of empowerment on client satisfaction and clinical outcomes. PMID- 12640953 TI - Clinical corner ... caffeine & women's health. PMID- 12640952 TI - Rheumatoid arthritis and the Arava (leflunomide) controversy. PMID- 12640954 TI - PA works toward state small pox plan. PMID- 12640955 TI - Nurse staffing and patient outcomes. This is news? PMID- 12640956 TI - Bedside placement of small-bowel feeding tubes in the intensive care unit. AB - Early enteral feeding is beneficial to critically ill patients, although it is often a challenge to accomplish. When required, placing feeding tubes into the small bowel can be costly and difficult to carry out in a timely manner, often requiring fluoroscopic or endoscopic guidance. We implemented a modified protocol that enabled nurses to place feeding tubes at the bedside without fluoroscopy. This technique has resulted in timely placement, with initiation of enteral feeding in less than 24 hours. Ninety-five percent of the attempted placements were successfully completed at the bedside. Transport of patients was avoided, and no specialized equipment was required. Therefore, greater satisfaction for patients with less risk was possible, and caregivers benefited as well. PMID- 12640957 TI - Reducing coronary artery disease by decreasing homocysteine levels. PMID- 12640958 TI - Managing nausea and vomiting. Current strategies. PMID- 12640959 TI - Use of nesiritide to treat acute decompensated heart failure. PMID- 12640960 TI - Early identification and management of critical incident stress. AB - Everyone experiences stress. That stress may be related to work (internal), community (external), or family; it may be cumulative or related to a particular critical incident. The cost related to treating acute stress is staggering, both to individuals and to organizations. Critical care nurses are well educated in the physiological responses to the stress of acute illness. Recognizing the emotional impact of stress and the techniques to manage it in themselves and in those with whom they work is equally as important. CISD is widely advocated as an intervention after critical incidents. Although debriefing in and of itself is effective, a single-session semistructured crisis intervention will not prevent posttraumatic stress; thus, the use of CISD as part of a comprehensive multifaceted approach to the management of acute stress related to a critical incident is recommended. PMID- 12640961 TI - Delirium during emergence from anesthesia: a case study. PMID- 12640962 TI - AACN Synergy model for patient care. Case study of a CHF patient. PMID- 12640963 TI - Nutrition support for the mechanically ventilated patient. AB - Nutrition support is a hotly debated topic in most intensive care units. Is enteral nutrition or TPN best? Is gastric or small-bowel feeding safer? Are specialized formulas needed? These are only some of the issues, and the fact remains that there is a paucity of clear, solid data. Folklore has become the standard of practice in many areas of medicine; it is richly found in nutrition support. We must be careful not to get caught up in the trappings of our beliefs about nutrition support. Instead, we must continue to evaluate our own practices and fine-tune our skills of clinical assessment and common sense. PMID- 12640964 TI - Measuring advanced practice nursing outcome, strategies and resources. PMID- 12640965 TI - New advances and novel treatments in heart failure. AB - As briefly summarized in this report, the prevalence of heart failure is high and it will continue to rise as the population ages. There will be over 1 million hospitalizations for acutely decompensated heart failure this year. The goals of treatment for patients with acutely decompensated heart failure are to lower cardiac filling pressures, remove fluids and improve symptoms of dyspnea, decrease vascular resistance, and increase cardiac output without activating the RAAS. There are few guidelines for the treatment of individuals with acutely decompensated heart failure and many different agents have been used in patients with this disease. Many of these drugs are not completely effective and may lead to serious adverse events. BNP is a natural protein produced by myocardial cells in response to ventricular distension, and its level is dramatically increased in patients with heart failure. The results of several recent clinical trials have shown that administration of nesiritide is safe and highly effective for the initial treatment of patients with acutely decompensated heart failure and can help physicians and nurses meet treatment goals for the management of patients with this serious condition. PMID- 12640966 TI - Who let the dogs out? Managing conflict with courage and skill. AB - Although nurses often think in terms of clinical ability when discussing competence, strong interpersonal communication skills are also essential for effective practitioners. Conflict management skills are key to creating work environments with good retention, work satisfaction, and high-quality measures. Nurses desiring to learn to communicate with courage and directness can master these skills, contributing to environments where caregivers can collaborate in providing the highest quality care. PMID- 12640967 TI - How nurse preceptors influence new graduates. PMID- 12640968 TI - AACN Synergy Model for Patient Care: a framework to optimize outcomes. PMID- 12640969 TI - BMP treatment for improving tendon repair. Studies on rat and rabbit Achilles tendons. PMID- 12640970 TI - [Genetic tests give the finishing touch to individual drug dosage]. PMID- 12640971 TI - [Pharmacogenetics--pathway to individualized antihypertensive pharmacotherapy]. AB - The variations in response to pharmacotherapy can in part be explained by genetic factors. Pharmacogenetics, a term coined in the 1950's, describes these relationships. Great technological advances in molecular biology and computational sciences, crowned by the sequencing of the human genome, have brought forth the current interest in pharmacogenetics. Continued technological developments may lead to the understanding of the molecular mechanisms behind complex diseases like cardiovascular disease, and holds the promise of individualized pharmacotherapy. The article gives a background to the field, with emphasis on antihypertensive therapy, and prospects for the future. PMID- 12640972 TI - [Medical education in Lund activates both students and teachers. Increased emphasis on general practice]. PMID- 12640974 TI - [Genetic research on blood samples stored for years in biobanks. Most people are willing to provide informed consent]. AB - Genetic research involving biobanks has been the subject of numerous declarations, recommendations and guidelines produced by professionals, administrators, legislators and other decision-makers. Yet, there is little published empirical information on people's willingness to participate in genetic studies on blood samples that they donated many years ago, at a time when the informed consent did not include the possibility of genetic research. We here report on our experiences of obtaining informed consent for academic and as well as commercial genetic research on blood samples collected more than a decade ago. Participants in a population-based risk factor survey who had donated blood to a biobank in 1990 were contacted eleven years later (in 2001) and asked for informed consent for genetic studies. A total of 1,311 out of 1,409 participants (93%) gave their consent to use blood samples for academic genetic research, provided that an Ethics Committee had approved the research. Thirty-one participants (2.2%) did not give their consent for academic genetic research. Another 35 subjects (2.5%) did not consent to industrial genetic research even if their blood samples were anonymized. Sixty-four individuals did not reply or provided incomplete answers (together 4.8%). Of the 1,311 individuals that agreed to participate, 292 (22.3%) wanted to be informed and give their concent to every new specific project. All the others gave a general concent provided that an Ethics Committee had approved the research. PMID- 12640973 TI - [Common, mostly in women and easy to treat. Primary hypothyroidism--an unknown public disease]. PMID- 12640975 TI - [In cases of coma and blue-green urine should overdosage of zaleplon be considered. First case reports]. PMID- 12640976 TI - [A twisted foot rescued Ferdinand Fagerlin from cholera]. PMID- 12640977 TI - [The proposal of the National Board of Health and Welfare counteracts knowledge development within health care]. PMID- 12640978 TI - [Private practitioners about Protos: the Medical Society fights for family practitioners' future!]. PMID- 12640979 TI - [New structure for patient-centered care is required]. PMID- 12640980 TI - [Researchers and psychiatrists defending Gillberg's research on ADDH: Karfve's campaign is a form of personal persecution and scientific basis is missing]. PMID- 12640981 TI - [Use continuous suture technique]. PMID- 12640982 TI - [Why there is no debate concerning the integrity-insulting ICD codes?]. PMID- 12640984 TI - [If the war against Iraq comes: Who will take the responsibility for five hundred thousand injured?]. PMID- 12640983 TI - [Setback for gene therapy: Two cases of leukemia in a French trial]. PMID- 12640985 TI - [Breaking bad news in oncology--more a matter of relation than information]. AB - Papers in clinical journals dealing with how to tell cancer patients bad news rely mostly on the opinion of the physician. The purpose of the present study was to contribute with knowledge from the patient's perspective by analysing how patients with recently diagnosed cancer narrate the manner in which they received their diagnosis. A consecutive series of 187 patients who had received their cancer diagnosis 2-8 months prior to the outset of this study were asked to describe the manner in which they learned of their diagnosis in writing. A crucial finding from the submitted 138 written narratives was that the participants often described experiences from the first contact with hospital staff to the end of their treatment, rather than as a single instance of diagnosis communication. The relational meaning of these experiences was obvious. From the patient's perspective, "bad news" reflects the process of being diseased by cancer, and how medical services are available when one is in need of establishing a helpful relationship. PMID- 12640986 TI - [Well-designed training programs crucial for dialysis patients]. AB - For many reasons patients on hemodialysis treatment have a low physical capacity. In this study the physical capacity was tested in 41 patients with chronic renal failure during their first weeks on hemodialysis and after 18 months of treatment. Compared to reference values the test result showed a significantly decreased physical strength, endurance and balance capacity already at the start of the dialysis. The patients were tested and got individually tailored information at the kidney unit and received some physical training outside the hospital. After 18 months the result for the group was not improved in spite of the fact that the 11 most frail patients had died. Diabetic and elderly patients had a significantly lower result than the rest of the patients. Future research will show, if more regular exercise (predialytic) individually programmed by a specially trained physiotherapist who can obtain up-to-date medical information as working in close contact to a dialysis unit, will further improve the condition of these patients. PMID- 12640988 TI - [A Scottish physician served as a model for Sherlock Holmes]. PMID- 12640987 TI - [Vascular agents can prevent type 2 diabetes]. PMID- 12640989 TI - [The governmental proposal on ethical review of research projects: manifestation of distrust concerning professional competence within universities and colleges]. PMID- 12640991 TI - [There are no special, fundamental reasons to say no to reproductive cloning]. PMID- 12640990 TI - [Final reply on bone density measurement in the heel: Diagnosis of osteoporosis should be done by DXA-measurement in the hip and spine]. PMID- 12640992 TI - [Unreasonably short time to considerate the proposal about the national family practice system]. PMID- 12640993 TI - [Doubtful re-evaluation of Roaccutane]. PMID- 12640994 TI - [Bone Marrow Transplantation-/Stem Cell Transplantation Congress of Nursing in Marburg: methods of support in extreme life circumstances]. PMID- 12640995 TI - [Otorhinolaryngology--4: Pro and contra of ear drops]. PMID- 12640996 TI - [Meal sharing in psychiatry: daily routine on forced responsibility]. PMID- 12640997 TI - [Re-sterilization of disposable products: a gamble]. PMID- 12640998 TI - [Development and implementation of public health plans: blind adoption is prohibited]. PMID- 12640999 TI - [Compliance with hand disinfection: evaluating intensive care units]. PMID- 12641000 TI - [Surface disinfection in the hospital: "Germany is number 1 in disinfection, yet still not a winner" (interview by Katrin Balzer)]. PMID- 12641001 TI - [In service learning in nursing care: sore points]. PMID- 12641002 TI - [Decubitus ulcer prevalence--a study of 11 health facilities: warning numbers]. PMID- 12641003 TI - [Nursing care of patients with HIV and AIDS: curriculum in the red ribbon symbol]. PMID- 12641005 TI - [Preparation for personal capital negotiation--5: Rating--communication with the bank]. PMID- 12641004 TI - [Viral hepatitis and HIV/AIDS: patients and personnel must be protected]. PMID- 12641006 TI - [Motivating co-workers: flexible control promises success]. PMID- 12641007 TI - [Life leadership: preserving balance in life]. PMID- 12641008 TI - [Memorandum regarding the Committee of Nursing Occupations]. PMID- 12641009 TI - Insights into vascular pathology after intracoronary brachytherapy. AB - Post-angioplasty restenosis is a major limitation of interventional cardiology. A large body of evidence reveals that expression of myofibroblast activity promoters moves progressively from the neoadventitia to the neointima. Brachytherapy inhibits vascular cell activity (proliferation, migration), mitigates recruitment of intimal cells, and shows a favorable prophylactic effect on late vascular remodeling by preventing adventitial constriction at the injured site. These effects of brachytherapy are dose related. Clinical and experimental data demonstrate that restenosis is determined by the balance between arterial remodeling and intimal hyperplasia. Apparently, brachytherapy-induced positive remodeling plays the principal role in increasing the luminal diameter after PTCA and, in case of a lower dose or dose fall-off, to cause detrimental edge effects. With regard to clinical course, healing defects, endothelial dysfunction and stent-vessel wall malapposition are apparently important and possibly underestimated features of vascular pathology, since they may contribute to late thrombosis and delayed intimal hyperplasia in long-term course after intracoronary brachytherapy. PMID- 12641010 TI - Validation of the localization of the target tissue for intracoronary brachytherapy. AB - In a previous meta-analysis of intracoronary brachytherapy (ICBT) studies, we identified the target tissue at 0.6 to 0.7 mm tissue depth and we developed two models, describing the relationship between dose and ICBT effectiveness. The purpose of the present study was to validate the identified target tissue depth and the developed dose models, using the results of 1) two prospective animal studies with ICBT, 2) a retrospective analysis of animal studies with external beam irradiation and 3) results of recent clinical ICBT trials. ICBT effectiveness in the porcine restenosis studies was quantified as inhibition of neointima proliferation. The results of these studies were correlated with the developed dose-effectiveness model. Finally, the agreement of the restenosis rates of the recent clinical trials with the developed dose-restenosis model was tested. The porcine restenosis studies demonstrated a dose-related inhibition of neointima proliferation. The radiation effectiveness of both prospective studies and the effectiveness of the studies with external beam irradiation demonstrated the best agreement with the developed dose model at a tissue depth of 0.6 mm. Furthermore, the restenosis rates of the recent clinical ICBT studies were in concordance with the developed dose-restenosis model. In conclusion, the current study validated the localization of the target tissue for ICBT at a tissue depth of 0.6 to 0.7 mm as well as the relationship between dose and ICBT effectiveness at this depth. The data provide a rationale for setting a common dose prescription point at 0.6 to 0.7 mm tissue depth. PMID- 12641011 TI - Serial intravascular ultrasound analysis after intracoronary beta-radiation in long in-stent restenotic lesions. AB - In-stent restenosis (ISR) represents the major limitation of stent implantation. Treatment, although of relative technical ease, is unsatisfactory due to the high incidence of recurrent restenosis. Long ISR lesions are especially prone to restenosis. Vascular brachytherapy (VBT) has emerged as a powerful adjunct therapeutic modality to treat ISR. However, VBT may be less effective in very long, diffuse ISR lesions. The present study investigated serial changes of the extent and distribution of neointima formation after beta-radiation as assessed by intravascular ultrasound (IVUS). Following interventional procedures of long ISR in 30 patients, a 40 mm or 60 mm non-centered 90Sr/90Y seed train was used for VBT. Serial (post radiation (PR) and follow-up (FU)) quantitative coronary angiography (QCA) measurements of minimal lumen diameter (MLD) and late lumen loss (LLL) and intravascular ultrasound measurements (IVUS) of cross-sectional areas of the lumen (L-CSA), stent (S-CSA) and intimal hyperplasia (IH-CSA) were performed and compared with historic controls. LLL (0.34 +/- 0.27 mm; p = 0.196), mean decrease of L-CSA (-1.0 +/- 0.8 mm2; p = 0.024) and mean increase of IH-CSA (0.5 +/- 1.3 mm2; p = 0.038) in long ISR were comparable with previously reported results of short ISR. In conclusion the average changes of lumen and intimal hyperplasia after beta-radiation of long ISR are similar to those of short ISR. PMID- 12641012 TI - Dose finding in intracoronary brachytherapy--consequences from empirical trials. AB - In-stent restenosis has been shown to be associated with a high recurrence rate of repetitive restenosis and remains a challenging task in interventional cardiology. Randomized, placebo-controlled trials have established that beta- as well as gamma-based vascular brachytherapy reduces the incidence of restenosis and clinical event rates following percutaneous coronary intervention (PCI) for the treatment of in-stent restenosis with focal and moderate length. Despite the number of clinical trials with impressive and convincing data, dose finding in most trials is empirical and remains an open question in this fairly new field of percutaneous interventional procedures. Current clinical trials have unequivocally demonstrated a clear dose dependency for the inhibition of intimal proliferation and a significant effectiveness for the treatment of in-stent restenosis with a dose around 20 Gy. Theoretical considerations and empirical data, however, support the need for a dose escalation with current systems to even further improve clinical results. A controlled dose escalation seems, thus, justified and is apparently not related with an increased risk of major adverse cardiac events. The current article gives an overview about theoretical considerations of dosing for intracoronary brachytherapy, presents recent data from important clinical trials in different views, and opens new perspectives for the successful treatment of in-stent restenosis. PMID- 12641013 TI - The IST registry. AB - Intracoronary brachytherapy has been established in Europe and the US as an evidence-based treatment of in-stent restenoses. The objective of the IST Registry is to register all patients treated in Germany with intracoronary radiation and to observe the clinical outcome for a duration of 5 years. The required set of data for each patient is kept to a minimum to encourage participation. All data are entered online. In the internet, each participating site can, at any time, check their most important parameters and compare them with those of other sites. Presently, the Novoste System is used in 58 catheter labs and the Guidant System in 16, while several sites use both. The requirements regarding radiation safety in intracoronary application of gamma radiation are very strict in Germany, so the Cordis-Gamma System is used in only one German lab. In a first analysis of 332 radiated stenoses, it was observed that late vessel occlusion could occur even after 6-month administration of clopidogrel (in addition to ASA)--without new stents being implanted within the brachytherapy session. Clopidogrel should thus be administered in addition to aspirin for at least a year. Ca. 270 patients per month receive intracoronary radiation in Germany, so the IST Registry will provide important data regarding long-term observation and a foundation for future negotiations with insurance companies potentially bearing the costs. At present, neither the physician's service nor the material costs are reimbursed. The IST Registry can furthermore be used as a comparative database regarding long-term outcome following implantation of antiproliferative-coated stents. PMID- 12641014 TI - Drug eluting stents: initial experiences. AB - Local delivery of immunosuppressive or antiproliferative agents using a drug eluting stent is a new technology meant to inhibit in-stent restenosis providing both a biological and mechanical solution and has recently emerged as a very promising approach. Up to now several agents have been in use: Paclitaxel, Rapamycin, Actinomycin D or Tacrolimus. Evaluating these drugs regarding their release kinetics, effective dosage, safety in clinical practice and benefit, several studies have been published or are still ongoing: SCORE (Paclitaxel derivative), TAXUS I, II, III, IV (Paclitaxel), ELUTE, ASPECT (Paclitaxel), RAVEL, SIRIUS (Sirolimus), ACTION (Actinomycin), EVIDENT, PRESENT (Tacrolimus). Paclitaxel was the first stent-based antiproliferative agent under clinical investigation providing profound inhibition of neointimal thickening, depending on delivery duration and drug dosage. The randomized multicenter SCORE trail (Quanam stent, Paclitaxel coated) enrolled 266 patients at 17 sites. At 6 month follow-up, a drop of 83% in stent restenosis using the drug-eluting stent could be achieved (6.4% drug-eluting stent vs. 36.9% control group) attributable to a remarkable decrease in intimal proliferation. Unfortunately, due to both frequent stent thrombosis and side-branch occlusions the reported 30-day MACE rate was 10.2%. The randomized TAXUS I safety trail (NIRx, Paclitaxel coated) also demonstrated beneficial reduction of restenotic lesions at 6-month FU (0% vs. 11%) but, this time, associated with the absence of thrombotic events presumably due to the lower drug dosage. The ongoing TAXUS II, III and IV trails are aimed at providing additional insight regarding the efficacy of the TAXUS Paclitaxel eluting stent. Both the RAVEL and the SIRIUS trial have been conducted to evaluate a Sirolimus-coated stent (Bx VELOCITY stent). From the results available, the beneficial findings regarding reduction of renarrowing using a drug-eluting stent have been confirmed without any adverse effects. Although parameters like drug toxicity, optimal drug dosage or delayed endothelial healing need to be further evaluated, summarizing the today's clinical experience the strategy of drug-coated stents promises a striking benefit in interventional treatment of coronary lesions. PMID- 12641015 TI - Sirolimus-eluting stents: a review of experimental and clinical findings. AB - Sirolimus (rapamycin), a macrolide antibiotic with known potent immunosuppressive properties, acts in the first phase (G1) of the cell cycle, blocking its further progression to the phase of DNA synthesis (S). In experimental models, rapamycin is effective in inhibiting smooth muscle cell proliferation and migration after vessel wall injury with balloon angioplasty. These results lead to the clinical application of sirolimus-eluting stents in 45 patients in Sao Paulo and Rotterdam (FIM Registry) and 238 patients in a randomized, European multicenter trial (RAVEL). These trials showed, by angiography and intravascular ultrasound, almost complete abolition of in-stent late hyperplasia up to one year after the procedure. In this review, we describe the experimental and clinical results of sirolimus-eluting stents including our experience of 26 stents implanted in 17 patients. In elective de novo lesions has shown remarkably clear lumens at follow up angiography and intravascular ultrasound within the stented segments were observed with no lesion progression at the stent margins or thrombosis after a 2 month regimen of aspirin, and ticlopodine or clopidogrel. New large-scale ongoing clinical trials will investigate the efficacy of sirolimus-eluting stents in lesions that are traditionally associated with high restenosis rates after stent implantation, such as long lesions, bifurcations and instent restenosis. PMID- 12641016 TI - Covered stent grafts: role in intervention of coronary arteries and degenerated vein grafts. AB - Covered stent grafts, metal stents covered by a polytetrafluoroethylene (PTFE) membrane, were developed in order to prevent restenosis, which is due to proliferation of tissue through the stent mesh. However, observational studies suggest that the cardiovascular event rate is elevated, when stent grafts are used electively in native vessels. In contrast, stent grafts appeared to be safe in the larger vessels of degenerated vein grafts. Potentially, microembolism of atherosclerotic debris might be reduced when used in saphenous vein grafts. Therefore, a randomized prospective study (STING) was performed to compare conventional stents with stent grafts in this indication. However, preliminary results do not support the hypothesis that stent grafts are superior. Further ongoing studies have to be awaited before a final conclusion can be drawn. Nevertheless, stent grafts are very helpful and reduce cardiovascular event rates and the need to perform emergency surgery in the case of a coronary rupture during a percutaneous intervention. Thus, having a stent graft available in the catheterization laboratory provides the interventionalist an important anchor for bail out situations. PMID- 12641017 TI - Clinical practice of primary angioplasty for the treatment of acute myocardial infarction in Germany: results from the MITRA and MIR registries. AB - The pooled data of two German AMI registries: the Maximal Individual Therapy in Acute Myocardial Infarction (MITRA) study and the Myocardial Infarction Registry (MIR) were analysed in order to 1) describe current clinical practice of primary angioplasty in Germany, 2) compare the results of primary angioplasty with those of thrombolysis in the "real world" and 3) define subgroups of patients profiting probably most from primary angioplasty. Between 1994 and 1998, 20,306 AMI patients were included in the registries. At the 271 participating hospitals angioplasty facilities were available at 18.5%. Thrombolysis was still the most frequently used reperfusion therapy at hospitals without (96%) as well as hospitals with such facilities (55%). Transfer of AMI patients for angiography was performed in 3.6% of AMI patients admitted to hospitals without angioplasty facilities. A total of 9906 lytic eligible AMI patients with a pre-hospital delay of no more than 12 hours were treated with either primary angioplasty (n = 1327) or thrombolysis (n = 8579). Univariate analysis of hospital mortality showed a more favourable course for patients treated with primary angioplasty: 6.4% versus 11.3%, OR = 0.54, 95% CI: 0.43-0.67, p < 0.0001. This was confirmed by logistic regression analysis: multivariate OR = 0.58, 95% CI: 0.44-0.77, p < 0.0001. Primary angioplasty was associated with a lower mortality in all subgroups analysed. There was a significant correlation between mortality and the absolute risk reduction (r = 0.82, p < 0.0001) in the different subgroups, which showed an increasing absolute benefit of primary angioplasty compared to thrombolysis with increasing mortality risk. PMID- 12641018 TI - Debulking prior to stenting--a worthwhile effort? AB - The rationale of debulking prior to stenting is to improve acute and long-term results of complex lesion angioplasty by reduction of plaque burden and/or reduction of calcified material prior to stenting, thus, allowing subsequent optimized stent-implantation and expansion. There are several registries analyzing the effect of various debulking-strategies prior to stenting in groups of selected patients and in high-risk subgroups. The majority of these studies performed DCA prior to stenting. In summary, these studies showed a reduction in restenosis rates and need for reintervention at long-term follow-up. The total TLR-rate (PCI, CABG) during a mean follow-up of 6 to 14 months is reported to be 4 to 14%; the rate of restenosis is given as 7 to 14%. However, until now, no large scale randomised "debulking prior to stenting" versus "conventional stenting without debulking" study has been published in detail. Several studies suggest benefit for debulking prior to stenting in complex lesion subsets, e.g., total chronic occlusions, ostial lesions, bifurcation lesions and saphenous vein grafts, but as of now, no convincing strategy has evolved. PMID- 12641019 TI - Transcatheter closure of intracardiac shunts. AB - In the last decade percutaneous transcatheter techniques and devices for closure of intracardiac defects have considerably improved. Transcatheter ASD closure has become a routine procedure in many centers. It is technically feasible in about 80% of the patients with a secundum type defect. The success rate today is higher than 99% and the rate of any complication below 0.5%. The advantages compared to surgery are the avoidance of a thoracotomy, postoperative pain and morbidity. The length of hospital stay is much shorter and most often general anesthesia is not necessary. Patent foramen ovale has been recognized as a potential cause of paradoxical embolism and embolic stroke particularly in younger patients. Transcatheter closure has recently gained popularity since several studies demonstrated a reduced rate of recurrent strokes after device closure. The occlusion is technically relatively simple and can be performed with ASD closure devices or with devices modified according to the anatomy of the foramen ovale. Catheter closure has certain advantages compared with life-long anticoagulation. For some patients this is the only therapeutic option. Nevertheless, there is still a need for randomized studies. Catheter closure of ventricular septal defects is possible today in selected patients. There are devices available specifically designed for muscular defects, for perimembraneous defects and subaortic ventricular septal defects. PMID- 12641020 TI - Myocardial perfusion after transcutaneous/percutaneous myocardial laser revascularization. AB - The effectiveness of transcutaneous and percutaneous myocardial laser revascularization in patients with endstage coronary artery disease and angina refractory to pharmacological therapy has been proved by various studies. Angiogenesis associated with an improvement of microcirculation and myocardial perfusion may be responsible for the reduction of angina and the improved physical performance. Myocardial perfusion studies published so far are compared to our own results utilizing positron emission tomography before and after percutaneous myocardial laser revascularization. PMID- 12641021 TI - Ten years after introduction of intravascular ultrasound in the catheterization laboratory: tool or toy? AB - Intravascular ultrasound (IVUS) represents the gold standard in the assessment of atherosclerotic disease. It has deeply affected our understanding of coronary artery disease and therapeutic strategies. We learned that a phenomenon described in pathologic series plays a very important role during life--that of vessel remodeling. The type of remodeling (positive or negative) cannot been assessed by angiography but is clinically relevant for optimal, procedural lumen gain. The modern stent implantation technique with a high balloon to artery ratio and high balloon pressure is the result of the IVUS finding that the majority of stents are not optimally deployed despite appropriate angiographic results. IVUS is essential in the clinical practice for the assessment of ambiguous angiography, left main stem disease, complex lesion and in-stent restenosis. Furthermore, IVUS has become an indispensable research tool in the analysis of new therapeutic strategies such as coronary brachytherapy. PMID- 12641022 TI - Influence of stent design and deployment technique on neointima formation and vascular remodeling. AB - A variety of different stent types is available for the treatment of coronary stenosis. However, in-stent restenosis remains the major limitation for the use of these devices. Intracoronary ultrasound (ICUS) in addition to coronary angiography provides precise measurements of coronary wall dimensions during stent implantation and of intimal hyperplasia during follow-up. The extent of coronary injury during stent implantation was shown to play an important role in neointima formation. It is characterized by endothelial exposure, intima laceration, and media permeation. Stent-induced coronary injury has been considered to depend on stent design and stent strut size with consecutive deep wall laceration. ICUS analysis showed a correlation between the stent design and the amount of neointimal tissue proliferation. The role of adventitial remodeling in the process of restenosis is discussed controversially. Post-procedural stent expansion may provoke adventitial remodeling. The stent design and stenting strategy determines the extent of peri- and post-procedural coronary injury. Post procedural coronary morphologic changes and changes of the stent geometry depend upon the stent design. Beside further modifications as the use of drug-eluting stents the decrease of stent-related vessel injury should be an important criterion for the development of future stent design. PMID- 12641023 TI - Intravascular sonotherapy for treatment of in-stent restenosis. AB - Intravascular sonotherapy has been introduced as a potential alternative to brachytherapy for prophylactic and therapeutic application in patients after coronary stent placement. Sonotherapy represents a non-ablative, non-thermal form of therapeutic ultrasound. The ultrasound catheter with diameter between 4 and 5 French carries up to 6 electronic elements operating currently at a frequency of 1.0 to 1.4 MHz. Animal studies have proven the efficacy of intravascular sonotherapy with respect to a reduction of neointimal ingrowth after stent placement. A first multicenter observational study in humans has documented that its application is safe and yields favorable results after coronary stenting, also in complex patient and lesion characteristics. This technique is currently under investigation in large prospective, multicenter, randomized studies performed in patients with de novo stenting or documented in-stent restenosis. PMID- 12641024 TI - Rationale of combined intracoronary pressure and flow velocity measurements. AB - The routine use of sensor-tipped guide wires for the assessment of functional coronary lesion severity has become widespread in the catheterization laboratory. The physiological parameters derived from pressure or flow velocity measurements, fractional flow reserve, relative and absolute coronary blood flow velocity reserve show a high agreement with noninvasive stress testing for myocardial ischemia. However, since these parameters are based on either intracoronary pressure or flow velocity, they do not investigate the hemodynamics of the coronary circulation completely, leading to ambiguous or conflicting outcomes. Only the use of simultaneously measured distal pressure and flow velocity will avoid any possible misinterpretation of the data. We have successfully used an index of stenosis resistance during maximal hyperemia to characterize the functional significance of a coronary stenosis. Recent developments in guide wire technology, currently used for research investigations, put these simultaneous measurements within clinical reach. PMID- 12641025 TI - Safety of FFR-based treatment strategies: the Munich experience. AB - Fractional flow reserve (FFR) as a new technique for physiological assessment of coronary stenoses could identify patients with CAD in whom the deferral of an intended PCI was more beneficial than performing the planned procedure. It is up to now unknown whether a FFR-based therapy stratification is also safe in patients with multivessel disease and complex coronary lesions. This study demonstrates in 71 symptomatic patients with predominantly multivessel disease and angiographically intermediate coronary lesions that patients do not benefit from PCI procedures in terms of overall survival, target vessel patency or clinical symptoms during 12 month follow-up if FFR is above 0.75. PMID- 12641026 TI - Simultaneous Doppler and pressure recordings to assess microvascular dysfunction in chronic total coronary occlusions--potential for recovery during follow-up. AB - BACKGROUND: Microvascular dysfunction (MD) is frequently observed in chronic total coronary occlusions (TCOs). This impairs the use of Doppler velocimetry for the guidance of PTCA. It is yet unknown to what extent microvascular dysfunction may recover during follow-up, and whether this may influence long-term outcome. METHODS: After recanalization and stenting of TCOs (duration > 4 weeks) in 56 patients, coronary flow velocity reserve (CFVR) and fractional flow reserve (FFR) were recorded. MD was defined as a CFVR < 2.0 in the absence of a significant epicardial lesion (FFR > or = 0.75). Patients were followed for 5.3 +/- 1.5 months with reassessment of CFVR, FFR, and of LV function during a repeat angiography. RESULTS: MD was observed in 50% after PTCA. The CFVR improved during follow-up from 1.93 +/- 0.62 to 2.35 +/- 0.69 (p < 0.001), with a decrease of baseline APV (31.0 +/- 13.6 cm/s to 25.7 +/- 15.0 cm/s; p = 0.022). At follow-up MD was observed less frequently (20%). In 20 of 36 patients with initially impaired LV function, the ejection fraction (LVEF) improved. The CFVR at baseline did not predict LV recovery. Target vessel failure (restenosis > 50% diameter stenosis) occurred in 43%. CFVR did not predict TVF, but FFR tended to be lower in patients with TVF. In 8 patients with FFR > or = 0.93 after recanalization, no TVF was required. CONCLUSIONS: MD improved in more than 50% with initially impaired MD after PTCA during a long-term follow-up. CFVR at baseline was not closely related to LV recovery. The high rate of TVF was not predictable by CFVR. The FFR may be more valuable for optimizing PTCA in TCOs than Doppler parameters. PMID- 12641027 TI - Early detection of myocardial microcirculatory disturbances after primary PTCA in patients with acute myocardial infarction: coronary blood flow velocity versus sestamibi perfusion imaging. AB - BACKGROUND: The extent of myocardial salvage after primary percutaneous transluminal coronary angioplasty (PTCA) in acute myocardial infarction (AMI) is variable and can not be predicted on the basis of vessel patency. The aim of this study was to evaluate the tissue salvage and the microvascular integrity after successful intervention in AMI by coronary blood flow velocity and sestamibi perfusion imaging. METHODS: Twenty-two patients (17 m, 5f; mean age 57 +/- 14 yrs.) undergoing primary PTCA and stent implantation for AMI were studied. 99mTc Sestamibi was injected intravenously before intervention and single photon emission computed tomography (SPECT) was performed immediately after successful reperfusion to determine the area at risk before PTCA due to the minimal 99mTc Sestamibi redistribution. Sestamibi SPECT was repeated 3 days and 6 months after AMI. Area at risk (%) was determined automatically by myocardial perfusion tomography (PERFIT) with the use of a multistage, 3D iterative inter-subject registration of patient images to normal templates (2SD) and myocardial salvage was calculated. Coronary flow velocity was measured using a Doppler-tipped guidewire in the infarct-related artery after successful completion of primary PTCA and in an angiographically normal reference vessel. Absolute coronary flow reserve (CFR) and relative CFR (rCFR) were calculated using hyperemic to basal average peak velocity. RESULTS: Despite successful reperfusion of the target vessel (TIMI grade III flow) CFR and rCFR remained impaired (1.8 +/- 0.9 and 0.77 +/- 0.21). Area at risk decreased significantly from 21 +/- 9% to 9 +/- 10% (p < 0.05) corresponding to 11 +/- 8% myocardial salvage. Acute CFR and rCFR showed no correlation with the area at risk before and after primary PTCA. The increase of CFR within 6 months correlated with the myocardial salvage (p < 0.05). CONCLUSIONS: Despite successful primary PTCA in AMI, CFR and rCFR often remain impaired because of a significant loss of microvascular integrity. The long-term success of primary PTCA can be assessed by myocardial salvage and the change of CFR which might be a useful parameter for additional reperfusion strategies such as glycoprotein IIb/IIIA receptor inhibition. PMID- 12641028 TI - Fractional flow reserve predicts major adverse cardiac events after coronary stent implantation. AB - OBJECTIVES: Determination of fractional flow reserve (FFR) allows the functional assessment of coronary stenoses before and after an intervention. Preliminary data suggest that a FFR > or = 0.94 is associated with an excellent clinical outcome after stent implantation. However, these results were limited both by the number of patients included and the use of non-contemporary stent designs. We sought to determine the prognostic value of FFR measurements in a large patient cohort undergoing coronary stent implantation. METHODS: Eighty-nine consecutive patients were enrolled in whom a stent implantation was performed and a pressure wire was used as a guide wire. Patients were followed for at least 6 months. Death, myocardial infarction (MI) and target vessel revascularization (TVR) were considered cardiac events. A FFR > or = 0.94 was regarded as an optimal functional result. RESULTS: A complete follow-up was available in all patents. Pre-interventional FFR increased from 0.66 +/- 0.16 to 0.95 +/- 0.05 (p < 0.0001) after stent implantation. Sixteen (18%) events occurred during follow-up including 10 (11.2%) TVR. Final FFR was significantly higher in patients without compared to patients with an event (0.92 +/- 0.06 vs. 0.96 +/- 0.05, p < 0.003). By univariate analysis, the presence of diabetes mellitus, left ventricular function, residual diameter stenosis and final FFR were associated with a worse clinical outcome. In the multivariate analysis, only the final FFR and left ventricular function remained as significant predictors for cardiac events (relative risk, 3.50; 95% CI: 1.29-9.52, P < 0.014, and 0.97; 95% CI: 0.93-1.00, p = 0.047). CONCLUSION: These results demonstrate in a nonselected patient cohort a strong correlation between FFR values after coronary stenting and subsequent cardiac events. Further studies have to investigate whether outcome after stenting might be improved by guiding the procedure with a pressure guide wire. PMID- 12641029 TI - Examining the economic impact of restenosis: implications for the cost effectiveness of an antiproliferative stent. AB - Over the past decade, coronary stenting has been shown to reduce the rates of angiographic and clinical restenosis compared with conventional balloon angioplasty. Despite these improved outcomes, however, coronary stenting is still hampered by a high incidence of restenosis, with important clinical and economic consequences. Recently, the development of antiproliferative, drug-eluting stents has emerged as a promising solution for the primary prevention of restenosis. This paper summarizes the current evidence on the economic impact of coronary restenosis and explores the potential impact of introducing an antiproliferative stent on the cost-effectiveness of percutaneous coronary revascularization. A decision-analytic model based on current clinical and cost data is used to examine the potential cost-effectiveness of such stents. Although prospectively designed studies will be critical to define the true impact of drug-eluting stents on long-term survival, quality of life, and costs in a broad patient population our decision analytic model suggests that as long as these stents are reasonably priced, they may be cost saving for certain patients and cost effective for virtually all patients undergoing PCI--at least within the U.S. healthcare system. PMID- 12641030 TI - The role of decision-analytic models in the prevention, diagnosis and treatment of coronary heart disease. AB - BACKGROUND: The rapidly expanding number and scope of health technologies used in the prevention, diagnosis, and treatment of coronary heart disease present physicians and policy makers with complex decisions that must be made even under uncertainty about the risks and benefits of these procedures. A physician must determine which procedures offer the best trade-off between potential harm and benefit, and yield the maximum expected health benefit for the patient. In addition, the cost-effectiveness of alternative choices must be considered to optimize resource allocations from a societal perspective, further complicating the decision making process. METHODS: Decision analysis is the application of explicit and quantitative methods to analyze decisions under conditions of uncertainty. Briefly, the basic concepts of decision analysis and cost effectiveness analysis are described. Three examples of decision-analytic models for the prevention, diagnosis, and treatment of coronary heart disease are discussed to demonstrate situations in which decision analysis may be helpful, and to introduce different methodological approaches. CONCLUSION: Decision analysis may aid clinical decisions affecting individual patients as well as inform clinical policy decisions and decisions regarding national health policy. However, this method is not a complete procedure for determining resource allocation decisions in health care, because it cannot incorporate all the values relevant to such decisions. PMID- 12641031 TI - If you were called upon in a tragedy would you be prepared. PMID- 12641032 TI - Hygiene--profit center or loss leader? PMID- 12641033 TI - Dentistry in colonial America. PMID- 12641035 TI - Oral pathology quiz #37. Case number 2. Primary squamous cell carcinoma. PMID- 12641034 TI - Oral pathology quiz #37. Case number 1. Ameloblastoma. PMID- 12641036 TI - Oral pathology quiz #37. Case number 3. Focal cemento-osseous dysplasia. PMID- 12641037 TI - Oral pathology quiz #37. Case number 4. Hyperplastic dental follicle. PMID- 12641038 TI - Thinking about things not thought of. Why it is important to volunteer and thank those who do. PMID- 12641039 TI - Tick talk. PMID- 12641040 TI - Collaboration and presence protect practice. PMID- 12641041 TI - Be part of nursing's solutions. Participate in nurse days at the capitol. PMID- 12641042 TI - What will solve the nursing shortage? PMID- 12641043 TI - Partnerships plan for potential smallpox. PMID- 12641044 TI - New safe bifurcated needle devices now available. PMID- 12641045 TI - Mercury thermometers pose problems. PMID- 12641047 TI - What is HIPAA privacy? PMID- 12641046 TI - Ban on artificial nails recommended for anyone having direct patient contact. PMID- 12641048 TI - [The political dimension of auxiliary personnel education: nursing and the unified health system (SUS)]. AB - The change in the administration of the municipality of Sao Paulo in 1989 innovated the management of health services aiming at implementing the Unified Health System (SUS) and using as a fundamental strategy the workers' qualification. Thus, the purpose of this work was to analyze the incorporation of the SUS principles in the programs for the capacitation of nursing auxiliary personnel. Based on the concepts that nurtured the Sanitary Reform and the construction of SUS principles, the methodological framework comprehended interviews with the coordinators of the program. Data analysis enabled authors to recognize that the municipal management policies and practices were committed to SUS guidelines, giving priority to the qualification of health workers who did not have this opportunity and adopting a pedagogy directed to the agents' transformation. The coordinators were motivated to develop the qualification program, recognizing this process as a tool for the SUS implementation. In addition, the coordinators at the regional level understood the technical dimension of this process, isolated from its political dimension. PMID- 12641050 TI - [Professional trajectory of obstetric nurses from the University of Sao Paulo College of Nursing: a focus on social phenomenology]. AB - This study aimed at understanding the experiences of students that were enrolled in the Obstetric Nursing Program offered by the University of Sao Paulo College of Nursing. Data were collected through interviews with the former students, who had different life histories after their graduation. The proposal of this research was based on Alfred Schutz' Social Phenomenology framework, considering that people express, in their actions socially experienced, the meaning of these experiences. Results showed two social types: the ones who continue in the area because they like what they do and the ones who do not work in the area as they were disappointed with the lack of professional autonomy. The reasons mentioned offer important guidelines that must be analyzed by the ones who are still working in the area, by the class associations and the ones who are responsible for the capacitation of these professionals. PMID- 12641049 TI - [Professionalization of nursing aides at the State of Sao Paulo: a study of supply/demand regarding training]. AB - The purpose of this exploratory study was to verify the problem of nursing aides professionalization and to analyze the offer of courses and demand regarding technical level capacitation at the state of Sao Paulo, Brazil. Data were obtained through the analysis of documents on professional capacitation in the area in the 1990s. Authors identified a scenario of conceptual and political changes regarding the professionalization and the existence of a great number of aides being professionalized. They also verified a tendency to increase the offer of courses and the demand regarding the capacitation and specialization at the technical level. PMID- 12641051 TI - The use of the "progressive muscle relaxation" technique for pain relief in gynecology and obstetrics. AB - Pain is one of the most frequent symptoms observed in patients and various treatments are proposed for its relief, including relaxation techniques. With the purpose of testing the effect of a specific intervention (progressive muscle relaxation) in a determined situation (pain), this study aimed at verifying the level of pain in post-surgery patients prior to and after the application of the Progressive Muscle Relaxation technique. The subjects, 61 patients, had been submitted to abdominal surgical interventions, of which 52.5% had a gynecological nature and 47.5% obstetrical. Our data showed statistically significant alterations in life parameters as well as muscular alterations after the application of the Progressive Muscle Relaxation technique. It was concluded that the use of the Progressive Muscle Relaxation Technique enabled the subjects to determine that their pain levels decreased. PMID- 12641053 TI - [Type of delivery: women's expectations]. AB - This descriptive and exploratory study was carried out through interviews with 221 puerperal women who gave birth in Sao Paulo State public maternities located within the capital. The purposes of the work were: 1. to identify the type of childbirth women expected to have--whether vaginal births or cesareans--and why; 2. to verify to what extent women's expectations corresponded to the type of birth they had; 3. to compare medical indications for c-sections with women's understanding of justifications they were given for this intervention. Data revealed that 74.7% of the women expected to have vaginal births and 25.3% expected to have cesarean sections. Vaginal birth, expected by 165 interviewees, occurred in 66.1% of these cases. Among women who expected having vaginal births, the most mentioned justification was that recuperation afterwards was faster. Among women who expected cesareans, the most mentioned justification was a previous c-section. The justifications presented by 61 women for having been submitted to c-sections did not coincide with medical indications for this intervention in 47.5% of the cases. PMID- 12641052 TI - [The elderly and asylums at the municipality of Campinas]. AB - This study aimed at characterizing the asylums at the municipality of Campinas regarding their residents, human resources and routines. Authors investigated six private institutions with residents, whose majority were dependent and with quantitatively and qualitatively insufficient human resources. It was possible to verify the capacitation of the health professionals in the areas of geriatrics and gerontology, in particular, as well as of the nursing team that is really important to the improvement of the care provided to the elderly. PMID- 12641054 TI - [The Saranson's Social Support questionnaire: studies regarding its adaptation to portuguese]. AB - With the aim of developing psychometrical studies using the Saranson's Social Support Questionnaire, an adapted version of the instrument was applied to 113 mothers. The Kappa Statistical Parameter, the Index of Gross Agreement and Spearman's Correlation were used to the analysis. The analysis of the internal consistency instrument showed a high level of it. In addition, the results indicated an acceptable test-retest reliability as well as statistical relationships between the social support indexes and variables related to work, pregnancy and income. Information on social support sources and the extension of the network suggest a greater participation of the family as the provider of the support. PMID- 12641055 TI - [Caring for chronic mental patients from the point of view of the nurse: a phenomenological focus]. AB - The purpose of this study, using the phenomenological framework, was to understand the phenomenon: caring about the care provided to the mentally ill patients in nurses' perspective. Therefore, authors interviewed nurses who work with mentally ill patients in two hospitals located at a city in the interior of the state of Sao Paulo, Brazil and also observed these places. In addition to the study with the aim of finding out nurse's view regarding the chronically ill patient, authors found out about nurse's perception about the care/work provided to these patients. Authors also reflected on caring, considering Heidegger's thought, what enabled them to better understand the phenomenon. PMID- 12641056 TI - [Evidence based practice: theoretical considerations for its implementation in perioperative nursing]. AB - Evidence-based practice enables the quality improvement of the nursing care delivered to clients/patients. This work aimed at providing elements that will lead to reflections on the scenario concerning perioperative nursing. Accordingly, based on literature, authors have made considerations regarding evidence-based nursing practice, general concepts, its importance and the obstacles to its implementation in professional practice, aiming at encouraging surgical nurses to seek for scientific knowledge by developing research and applying its results. PMID- 12641057 TI - [Reflexive education in post-modernity: a bibliographic review]. AB - This is a bibliographical review that aimed at considering teaching at the actual historical moment, denominated post-modernity, and that has resulted in a pragmatic change in all levels of human being understanding. Thus, authors rescued and presented the presuppositions for a post-modern education, emphasizing among them the reflexive education proposed by Donald A. Shon, that is based on practical learning centered in "reflexion-in-action", as an alternative for the formation of a reflexive professional. PMID- 12641058 TI - [Informed management in nursing work]. AB - Informed management presupposes that all individuals will prioritize a taming and determined conduct in detriment of a passive one. It also presupposes deep concern about issues related to the human species, ethical problems, the relationship with nature and, finally, about men's future. In this way, the authors, based on the assumptions underlying the Policies of Informed Management proposed by Maslow, aim at focusing on this type of management in nurses' work. They believe that, by means of such reflection, nurses will be able to incorporate new values which are more harmonized with human nature. PMID- 12641059 TI - [Pediatric nursing and the family approach: contribution to undergraduate teaching]. AB - This work aims at presenting the instruments used in the family nursing approach as depicted in the international literature as well as at providing subsidies for their utilization by Brazilian nursing professionals in family health approaches to child care. The authors present the experience of introducing a methodology for systematized care in Pediatric Nursing programs using instruments for data collection such as the family APGAR, Genogram and Ecomap. They argue that these instruments do not replace the theoretical basis of care or the reason and discernment in order to provide data with meaning and find better solutions to care for children and their family. PMID- 12641060 TI - [Adjustment of the theory of self-care deficit in home care in the light of Barnum's model]. AB - The study aimed at analyzing the Self-Care Deficit Theory adjustment, focusing on the atmosphere and the home care provider. Authors adopted the Barnum's model that suggests questions on the use of its concepts, seeking for its interrelations and scope. The answers to those questions were based on the Self Care Deficit Theory applied to a real situation, observed in home care visits occurred in the second semester of 2000 to a paraplegic patient, whose care was provided by this wife and the family health nursing team. Authors found minimal adjustment conditions for home care, considering the environment and the care providers. They considered, comparatively, that the chosen theory may be directed to the care of patients in home hospitalization under the supervision of a nurse. PMID- 12641061 TI - [Tutorial programs for nursing courses]. AB - The implementation of political-pedagogical projects, encouraged by health and education policies, generated the need to incorporate in nursing curricula several programs to attend the interests and necessities of undergraduate courses. One of these is the Tutorial Program that aims at considering the student as the subject of the education and care. In this way, this study aimed at contributing to the evaluation of a tutorial model in order to fill in gaps found in the process of construction of political-pedagogical projects. Therefore, the article describes the idealized theoretical model with the purpose to include it in undergraduate nursing programs. PMID- 12641062 TI - [Care for parturient women by obstetrical nurses in the Midwifery project: an experience report]. AB - This study focuses on the nursing care models that value the woman in childbirth process. Authors described the humanized care provided to the parturient by obstetric nurses in the "Midwifery Project" developed at a University Maternity Hospital. They emphasized that the practice of these models enable the nurse's active participation in the childbirth process, resulting in the satisfaction of the parturient and of the professional. PMID- 12641063 TI - Peer mentorship: a crucial ingredient for success in nursing school. PMID- 12641064 TI - Support the uninsured March 10-16 is "cover the uninsured" week. PMID- 12641065 TI - Advanced practice nursing. Options and opportunities. PMID- 12641066 TI - A career in nurse anesthesia. PMID- 12641067 TI - The clinical nurse specialist. PMID- 12641068 TI - Nurse midwifery. PMID- 12641069 TI - An old tool for a new job market. PMID- 12641070 TI - 12 frequently asked questions about the NCLEX-RN. PMID- 12641072 TI - [Abstracts of the VIII Ukrainian Biochemistry Congress. October 1-3. Chernivtsi]. PMID- 12641071 TI - [Abstracts of the VIII Ukrainian Biochemical Congress. October 1-3, 2002. Chernivtsi]. PMID- 12641073 TI - Psychological coping and cancer. Search strategy used is inadequate. PMID- 12641074 TI - Reconfiguring health systems. Hospital buildings and increasing specialisation are here to stay. PMID- 12641075 TI - Effectiveness of trained nurses in preoperative assessment. Paper is missing the point of preoperative assessment. PMID- 12641076 TI - Mind and body split. Medical professionals in Asia have subscribed to Western thought. PMID- 12641079 TI - The American College of Cardiology (ACC)--51st Annual Scientific Session. PMID- 12641080 TI - Cardiostin 2002. PMID- 12641081 TI - Heart failure 2001. Fourth international meeting organized by the Working Group on Heart Failure of the European Society of Cardiology. PMID- 12641082 TI - Undocumented patient information: an impediment to quality of care. AB - PURPOSE: Poor documentation in medical records might reduce the quality of care and undermine analyses based on retrospective chart reviews. We assessed the documentation of cardiac risk factors and cardiac history in the records of patients hospitalized with myocardial infarction or heart failure. METHODS: We performed a retrospective cohort study involving direct chart audit of all consecutive hospitalizations for myocardial infarction (n = 2,109) or heart failure (n = 3,392) in Nova Scotia, Canada, from October 15, 1997, to October 14, 1998. The main outcome measures were the documentation rates for prespecified clinical items, including cardiac risk factors and history of myocardial infarction or heart failure, which were recognized as indicators of the quality of care for the conditions under study. RESULTS: Information was not documented in a high proportion of cases, ranging from 9% (smoking) to 58% (previous history of heart failure) in charts from patients hospitalized for myocardial infarction, and from 19% (smoking) to 69% (hyperlipidemia) in charts from heart failure hospitalizations. Lack of documentation was more common in women and the elderly. CONCLUSION: Documentation of important clinical information is poor even in the hospital charts of patients with severe conditions. This quality-of-care issue has implications for health services and outcomes research, including the development of report cards. PMID- 12641083 TI - Re: Bilateral laparoscopic inguinal hernia repair can complicate subsequent radical retropubic prostatectomy. PMID- 12641084 TI - A randomized trial comparing radical prostatectomy with watchful waiting in early prostate cancer. PMID- 12641085 TI - Quality of life after radical prostatectomy or watchful waiting. PMID- 12641087 TI - A novel founder mutation in the RNASEL gene, 471delAAAG, is associated with prostate cancer in Ashkenazi Jews. PMID- 12641086 TI - Germline mutations and sequence variants of macrophage scavenger receptor 1 gene are associated with prostate cancer risk. PMID- 12641088 TI - RNASEL Arg462Gln variant is implicated in up to 13% of prostate cancer cases. PMID- 12641089 TI - Serum proteomic patterns for detection of prostate cancer. PMID- 12641091 TI - Dietary supplement use in the Prostate Cancer Prevention trial: implications for prevention trials. PMID- 12641090 TI - Lack of effect of a low-fat, high-fruit, -vegetable, and -fiber diet on serum prostate-specific antigen of men without prostate cancer: results from a randomized trial. PMID- 12641092 TI - Comparison of the efficacy of local therapies for localized prostate cancer in the prostate-specific antigen era: a large single-institution experience with radical prostatectomy and external-beam radiation. PMID- 12641093 TI - On the edge: low-wage workers and their health insurance coverage. Findings from the Commonwealth Fund 2001 Health Insurance Survey. PMID- 12641094 TI - Defining sexual outcomes after treatment for localized prostate carcinoma. PMID- 12641096 TI - (In)-efficacy of salvage radiotherapy for rising PSA or clinically isolated local recurrence after radical prostatectomy. PMID- 12641095 TI - Postoperative radiotherapy in 423 patients with pT30NO prostate cancer. PMID- 12641098 TI - Post-nerve-sparing prostatectomy, dose-escalated intensity-modulated radiotherapy: effect on erectile dysfunction. PMID- 12641097 TI - Process involved in reading imaging studies: workflow analysis and implications for workstation development. AB - Software development for imaging workstations has lagged behind hardware availability. To guide development and to analyze work flow involved in interpretation of cross-sectional imaging studies, we assessed the cognitive and physical processes. We observed the performance and interpretation of body computed tomography (CT scans and recorded the events that occurred during this process. We studied work flow using a bottleneck analysis. Twenty-four ofa total of 54 cases (44%) involved comparing the images with those of prior scans. Forty seven of 54 scans (87%) were viewed using windows other than soft tissue, or compared with precontrast scans. In 46 cases (85%), the interpretation stopped to return to a previous level for review. Measurement of lesions was performed in 24 of 54 (44%) cases, and in 15 (63%)of these cases, measurements were taken of lesions on old studies for comparison. Interpretation was interrupted in 14 of 54 cases (26%) by referring clinicians desiring consultation. The work flow analysis showed film folder retrieval by the film room to be the bottleneck for interpretation by film. For picture archiving and communication system (PACS) reading,the CT examination itself proved to be the bottleneck. We conclude that workstations for CT interpretation should facilitate movement within scans, comparison with prior examinations, and measuring lesions on these scans. Workstation design should consider means of optimizing time currently not used between interpretation sessions, minimizing interruptions and providing more automated functions currently requiring physician interaction. PMID- 12641099 TI - Predictive factors of urinary retention following prostate brachytherapy. PMID- 12641100 TI - The impact of androgen deprivation on quality of life after radical prostatectomy for prostate carcinoma. PMID- 12641101 TI - Altered cognitive function in men treated for prostate cancer with leutinizing hormone-releasing hormone analogues and cyproterone acetate: a randomized controlled study. PMID- 12641102 TI - Renal angiomyolipoma: relationships between tumor size, aneurysm formation, and rupture. PMID- 12641103 TI - Tumors and tumorlike lesions of the testis: radiologic-pathologic correlation. PMID- 12641104 TI - Imaging for nephron-sparing surgery. PMID- 12641106 TI - Exstrophy of the cloaca and exstrophy of the bladder: two different expressions of a primary development field defect. PMID- 12641105 TI - Results of systematic voiding cystourethrography in infants with antenatally diagnosed pelvis dilation. PMID- 12641107 TI - Is an artificial sphincter the best choice for incontinent boys with Spina Bifida? Review of our long term experience with the AS-800 artificial sphincter. PMID- 12641108 TI - High-pressure bladder: an underlying factor mediating renal damage in the absence of reflux? PMID- 12641110 TI - A long-term follow-up of autoaugmentation in myelodysplastic children. PMID- 12641109 TI - Detrusor myotomy: a 5-year review in unstable and non-compliant bladders. PMID- 12641111 TI - Endocrine-related resources from the National Institutes of Health. PMID- 12641112 TI - The outcome of voiding dysfunction managed with clean intermittent catheterization in neurologically and anatomically normal children. PMID- 12641113 TI - Cremasteric reflex and retraction of a testis. PMID- 12641115 TI - Ambiguous genitalia with perineoscrotal hypospadias in 46,XY individuals: long term medical, surgical, and psychosexual outcome. PMID- 12641114 TI - 46,XY intersex individuals: phenotypic and etiologic classification, knowledge of condition, and satisfaction with knowledge in adulthood. PMID- 12641116 TI - Medical photography: ethics, consent and intersex patient. PMID- 12641117 TI - Conversations with Robert Truax. Interview by J.R. Wilson. PMID- 12641118 TI - Periprostatic nerve block gives better analgesia for prostatic biopsy. PMID- 12641119 TI - Perioperative changes in serum creatine after gynecologic surgery. PMID- 12641120 TI - Laparoscopic radical cystectomy with urinary diversion: completely intracorporeal technique. PMID- 12641121 TI - Urethra-sparing cystectomy and orthotopic urinary diversion in women with malignant pelvic tumors. PMID- 12641123 TI - Treatment of benign ureterointestinal anastomatic strictures with permanent ureteral Wallstent after Camey and Wallace urinary diversion: long-term follow up. PMID- 12641122 TI - Long-time experience with Kock ileal reservoir for continent urinary diversion. PMID- 12641125 TI - Ten years' experience with submucosally embedded in situ appendix in continent cutaneous diversion. PMID- 12641124 TI - Health-related quality of life after radical cystectomy for bladder cancer: a comparison of ileal conduit and orthotopic bladder replacement. PMID- 12641126 TI - Ileal T-pouch as a urinary continent cutaneous diversion: clinical and urodynamic evaluation. PMID- 12641127 TI - A high easy-to-treat complication rate is the price for continent stoma. PMID- 12641128 TI - Asymptomatic bacteriuria in elderly humans is associated with increased levels of circulating TNF receptors and elevated number in neutrophils. PMID- 12641129 TI - Geysers spawn hot new nanotechnology. PMID- 12641130 TI - Global molecular epidemiology of the O15:K52:H1 extraintestinal pathogenic Escherichia coli clonal group: evidence of distribution beyond Europe. PMID- 12641131 TI - Molecular epidemiology of 3 putative virulence genes for Escherichia coli urinary tract infection--usp, iha, and iroN(E. coli). PMID- 12641132 TI - Columbia--aftermath of a tragedy. PMID- 12641133 TI - Identification of target tissue glycosphingolipid receptors for uropathogenic, F1C-fimbriated Escherichia coli and its role in mucosal inflammation. PMID- 12641134 TI - Refuse-to-crash: NASA tackles loss of control. PMID- 12641135 TI - Non-secretor status is associated with female acute uncomplicated pyelonephritis. PMID- 12641136 TI - Short compared with standard duration of antibiotic treatment for urinary tract infection: a systematic review of randomised controlled trials. PMID- 12641137 TI - Antibiotic treatment for five days is effective in children with acute cystitis. PMID- 12641138 TI - Utility of blood cultures in febrile children with UTI. PMID- 12641139 TI - Urinary tract infection as a possible marker for teenage sex. PMID- 12641140 TI - National Institutes of Health Chronic Prostatitis Symptom Index for Japanese men. PMID- 12641141 TI - Cytologic evaluation of urine is important in evaluation of chronic prostatitis. PMID- 12641142 TI - Transurethral needle ablation for the treatment of chronic pelvis pain syndrome (category III prostatitis): a randomized, sham-controlled study. PMID- 12641143 TI - Evaluation of the DipStreak, a new device with an original streaking mechanism for detection, counting, and presumptive identification of urinary tract pathogens. PMID- 12641144 TI - Office-based transurethral microwave thermotherapy using TherMatrx TMx-2000. PMID- 12641145 TI - Association of polymorphisms within androgen receptor, 5 alpha-reductase, and PSA genes with prostate volume, clinical parameters, and endocrine status in elderly men. PMID- 12641146 TI - Boosted decision tree analysis od surface-enhanced laser desorption/ionization mass spectral serum profiles discriminates prostate cancer from noncancer patients. PMID- 12641147 TI - The value of power Doppler imaging to predict the histologic components of benign prostatic hyperplasia. PMID- 12641148 TI - Expression of somatostatin receptor subtypes 2 and 4 in human benign prostatic hyperplasia and prostatic cancer. PMID- 12641149 TI - Effects of intrathecal injection of tamsulosin and naftopidil, alpha-1A and 1D adrenergic receptor antagonists, on bladder activity in rats. PMID- 12641150 TI - Quantitative evaluation of prostatectomy for benign prostatic hypertrophy under a national health insurance law: a multi-centre study. PMID- 12641151 TI - Increased bladder apoptosis with alpha-1 adrenoceptor antagonists in benign prostatic hyperplasia. PMID- 12641152 TI - The nursing shortage--a reason for concern? PMID- 12641153 TI - The creation of formal planning guidelines for newborn intensive care units (NICUs). PMID- 12641154 TI - "Cold" solid-phase microextraction method for the determination of volatile halocarbons present in the atmosphere at ultra-trace levels. AB - Anthropogenic volatile halocarbons are compounds of great enviromnental concern because of their involvement in global change phenomena. They are present in the atmosphere at concentration levels in the order of parts per trillion by volume. The chosen analytical method for their determination is capillary gas chromatography coupled to mass spectrometry, preceded by an enrichment step on suitable adsorbent resins. The method here presented makes use of the solid-phase microextraction as a pre-analytical technique, using sub-ambient temperature in order to enhance the retention capability of the fiber coating. The proposed method was evaluated in terms of extraction efficiency, linearity, reproducibility, andlimits of detection. Results obtained showed that trace atmospheric halocarbons are detectable even when enriching very small air sample volumes. A good chromatographic resolution is obtained as a consequence of the extremely low injection volume. Finally a standard GC-MS instrumentation equipped with a simple split-splitless injector was employed, thus avoiding the use of expensive dedicated apparatus. The method was also applied to the analysis of actual samples collected both in remote, and in semi-remote sites. PMID- 12641155 TI - Competitive extraction of multi-component contaminants in water by Carboxen polydimethylsiloxane fiber during solid-phase microextraction. AB - A headspace analysis for groundwater contaminated with BTEX (benzene, toluene, ethylbenzene, and xylenes) was employed to investigate the feasibility and limitations of Carboxen-PDMS (polydimethylsiloxane) fiber during SPME (solid phase microextraction). Although the response of the Carboxen-PDMS fiber was much higher than that of conventional PDMS fiber, a reduction of the extracted amount was also observed under multi-component conditions due to competitive replacement. The general affinity of analytes to the fiber was high in the order xylene>ethylbenzene>toluene> benzene. The behavior of the Carboxen-PDMS fiber was examined more precisely at constant compositional ratio, because the analysis of contaminants using Carboxen-PDMS fiber was reported to be possible at known composition. The relative affinity of each component was shown to differ according to the total amount of analytes. Furthermore, the extracted amounts of benzene and toluene did not show a consistent tendency as the concentration of each component increased. These results indicate that caution should be exercised if Carboxen-PDMS fibers are used for the analysis of BTEX in groundwater samples. PMID- 12641156 TI - Determination of single component isotherms and affinity energy distribution by chromatography. AB - Adsorption isotherm data were acquired by frontal analysis (FA) and large sample size band profiles were recorded for phenol and caffeine. For both compounds, the isotherm data fit well to the Langmuir, Toth, and Bi-Langmuir models of adsorption. The Langmuir model must be dismissed because it does not predict accurately the overloaded band profiles. However, profiles calculated using the unimodal Toth and the bimodal Bi-Langmuir models are indistinguishable. The expectation-maximization procedure was used to calculate directly the affinity energy distribution (AED) from the raw FA data points. For both compounds, the AED converges to a bimodal distribution at high numbers of iterations. This result, which shows the high sensitivity of the EM method, suggest that the Bi Langmuir model makes better physical sense than the Toth model. This model also permits a detailed investigation of the properties of active sites, a feature often evoked in chromatography but so far rarely the topic of a quantitative investigation. PMID- 12641157 TI - Influence of pressure on the chromatographic behavior of insulin variants under nonlinear conditions. AB - The effect of pressure on the chromatographic behavior of two insulin variants in RPLC was investigated on a YMC-ODS C18 column, under nonlinear conditions. The adsorption isotherm data of porcine insulin and Lispro were measured at average column pressures ranging from 52 to 242 bar. These data fit well to the Toth and the bi-Langmuir isotherm models. The saturation capacity increases rapidly with increasing pressure while the affinity (or equilibrium) constant and the parameter characterizing the surface heterogeneity decrease. It is noteworthy that the distribution coefficient of the insulin variants increases with increasing pressure whereas their equilibrium constant b decreases for porcine insulin and increases for Lispro. The association constant b(ds), which characterizes the adsorption and desorption equilibrium of insulin in the system, increases with increasing pressure. The excellent agreement between the experimental overloaded profiles recorded under different pressures and those calculated using the POR model suggests that the chromatographic behavior of insulin is controlled more by equilibrium thermodynamics than by the mass transfer kinetics. The latter seems to be nearly independent of the average column pressure. Thus, increasing the average column pressure is an efficient, albeit costly, way to increase the loading capacity of the column, hence the production rate in preparative chromatography. PMID- 12641158 TI - Modeling of preparative chromatography processes with slow intraparticle mass transport kinetics. AB - Mathematical modeling of the preparative chromatography process accompanied with complex intraparticle mass transport mechanism involving surface diffusion is discussed. As an experimental base for the analysis two steroid compounds, methyl esters of hydroxycholanic acids (bile acids), deoxycholic and cholic acids were selected. For these compounds surface diffusion kinetics were found to have a marked influence on the band broadening. The isocratic chromatography process was performed in a normal-phase preparative system with ternary mixture of solvents containing hexane, ethyl acetate and methanol as a modifier under different operating conditions, e.g., at various mobile phase compositions and inlet concentrations. The efficiency of the system was found to be dependent on the mass of sample injected as well as on the contents of the modifier. Such a phenomenon was suggested to originate from the contribution of the surface diffusion kinetics to the overall mass transport mechanism. For identifying the general trends and concentration dependencies of the surface diffusion coefficient the simplified approach was proposed. The set of chromatographic band profiles registered at different inlet concentration and mobile phase composition were used for determining the influence of the local solid-phase concentration on the mass transport mechanism. For the simulations the transport-dispersive model was used, in which all sources of mass transport resistances were lumped in the properly adjusted mass transport coefficient. The accuracy of this model was verified by comparing its predictions to the solutions of the general rate model. PMID- 12641159 TI - Parcel model for peak shapes in chromatography numerical verification of the temporal distortion effect to peak asymmetry. AB - The traditional plate concept has been reassessed and improved to a parcel matrix model, which can be used to imitate the chromatographic behavior of a hypothetic column on a computer worksheet. Under programmed conditions, various peak shapes (nearly Gaussian, and with prolonged or fronting tails) are generated. The peak tailing has been separated into two major fractions: spatial and temporal. The former fraction is caused by the retention nature of a column, whereas the latter is induced by the observer's relative position and the changing of the zone broadening rate. The temporal distortion effect can be identified qualitatively and quantitatively through a normalized peak-overlapping process. In general, a chromatographic peak may carry a prolonged (or normal type) tail under linear isotherms, while both prolonged and fronting tails will appear under non-linear conditions. The temporal distortion is proved to be significant, and may be regarded as the major cause of peak asymmetry in most cases. This is in contrast to the conclusions of many previous studies. The model is also eligible to simulate chromatographic peaks for various injection sizes. PMID- 12641160 TI - Classification and regression tree analysis for molecular descriptor selection and retention prediction in chromatographic quantitative structure-retention relationship studies. AB - The use of the classification and regression tree (CART) methodology was studied in a quantitative structure-retention relationship (QSRR) context on a data set consisting of the retentions of 83 structurally diverse drugs on a Unisphere PBD column, using isocratic elutions at pH 11.7. The response (dependent variable) in the tree models consisted of the predicted rention factor (log kw) of the solutes, while a set of 266 molecular descriptors was used as explanatory variables in the tree building. Molecular descriptors related to the hydrophobicity (log P and Hy) and the size (TPC) of the molecules were selected out of these 266 descriptors in order to describe and predict retention. Besides the above mentioned, CART was also able to select hydrogen-bonding and molecular complexity descriptors. Since these variables are expected from QSRR knowledge, it demonstrates the potential of CART as a methodology to understand retention in chromatographic systems. The potential of CART to predict retention and thus occasionally to select an appropriate system for a given mixture was also evaluated. Reasonably good prediction, i.e. only 9% serious misclassification, was observed. Moreover, some of the misclassifications probably are inherent to the data set applied. PMID- 12641161 TI - Using second-order calibration to identify and quantify aromatic sulfonates in water by high-performance liquid chromatography in the presence of coeluting interferences. AB - We used the Generalized Rank Annihilation Method (GRAM), a second-order calibration method, to quantify aromatic sulfonates in water with high performance liquid chromatography (HPLC) when interferences coeluted with the analytes of interest. With GRAM, we can quantify in only two chromatographic analyses, one for a calibration sample and one for the unknown sample. The calculated concentrations were not statistically different to those obtained when the chromatographic separation of the unknown sample was modified in order to completely separate the analyte from the interferences before univariate calibration. With GRAM, the concentrations are determined much more quickly because a complete resolution is not required. PMID- 12641162 TI - Lowering the molecular mass limit of thermal field-flow fractionation for polymer separations. AB - Thermal field-flow fractionation (ThFFF) is capable of separating a wide molecular mass range of polymers by their molecular mass (Mr) and chemical composition. However, retention and resolution decrease significantly for polymers with Mr<20 kDa. Various approaches for increasing the retention of low Mr (<15 kDa) polymers were investigated. Our results showed that temperature conditions and single-component solvents had a limited effect on polymer retention and that certain binary solvent mixtures caused a dramatic increase in retention. The binary solvents approach has enabled the use of a ThFFF system and temperature conditions to separate 2.6 kDa PS from 4.4 kDa PS, thereby extending the applicability of ThFFF to lower molecular masses. The effect of binary solvent mixtures on polymer retention is correlated with the mixture viscosity. PMID- 12641163 TI - Systematic development and validation of stability indicating micellar electrokinetic chromatography methods for early stage drug candidates. AB - Suitable methods for assessment of purity, potency and stability of new drug substances and drug products are required to be rapidly developed and validated to provide appropriate data for early project development decisions. In order to routinely provide methods of consistent and suitable quality to meet increasingly aggressive timelines, systematic approaches to both develop and validate analytical technologies have been developed. Systematic approaches to evaluate separation parameters such as buffer pH, buffer ionic strength, surfactant concentration, organic modifiers, organic modifier concentration, applied voltage and temperature were evaluated for an early stage drug candidate. Techniques to improve method precision and ruggedness were also examined. Finally, the validation results from the micellar electrokinetic chromatography method utilizing an internal standard were compared against the simultaneously developed high-performance liquid chromatography method. PMID- 12641165 TI - Dental postcards XXIII. At the Dentist. PMID- 12641164 TI - Explanation for the enhanced dissolution of silica column packing in high pH phosphate and carbonate buffers. AB - It has been reported that at high pH, the rate of bonded phase packing degradation in methanol/water mobile phases is greater for carbonate and phosphate buffers than for amine buffers. This conclusion was based on buffer pH determined in the aqueous buffer before dilution with methanol. Changes in buffer species pKa, and therefore buffer pH, upon methanol dilution are consistent with the observed degradation results. Measurements of pH in the methanol/water solutions confirm that the carbonate and phosphate buffers were considerably more basic than the amine buffer, even though all the buffers were pH 10 before dilution with methanol. These results demonstrate that it can be misleading to extrapolate aqueous pH data to partially aqueous solutions. Measurements of pH in the mixed solvent provide more reliable predictions of column and sample stability. PMID- 12641166 TI - The 1850 Webster/Parkman Trial: Dr. Keep's forensic evidence. AB - Shortly before two o'clock on a chilly November afternoon in 1849, the celebrated Harvard physician and surgeon, Dr. George Parkman, left his home on Boston's fashionable Beacon Hill, expecting to return in a few hours. He was never seen alive again. This account describes Parkman's brutal murder and explores the dynamics which preceded this crime. It explains how and why Dr. John White Webster, MD, Professor of Chemistry and Mineralogy at Harvard University, killed Dr. Parkman and unsuccessfully attempted to destroy the physical evidence. Webster's subsequent trial, conviction and ultimate punishment are also detailed. The Parkman-Webster case remains one of the classic murders in the annals of American crime. Compelling dental evidence presented by the famous American dentist, Dr. Nathan Cooley Keep, directly led to the conviction of Dr. Webster. This graphic, ground-breaking case clearly established the viable role of forensic dentistry in legal criminal investigation. PMID- 12641167 TI - Gleanings about dentistry from the world of literature (twenty-ninth in a series). PMID- 12641168 TI - Levi Spear Parmly: father of dental hygiene and children's dentistry in America. AB - Levi Spear Parmly (1790-1859) was one of the outstanding dental practitioners and teachers in the early part of the eighteenth century, both in America and Europe. He published two highly regarded books, practiced and taught in London for several years, before returning to America, where he became the most eminent dental practitioner in the southern United States. He had great financial success and used his time and money to offer free dental service to children. His major contribution to dental science was the concept that dental caries was caused by external influences related to foreign material on the tooth surface. This was before our knowledge of dental plaque and the action of the oral bacteria. Based upon this concept, he stressed the importance of clean teeth to prevent tooth decay. PMID- 12641169 TI - The triumph and tragedy of James Baxler Bean, MD, DDS (1834-1870). AB - In 1863, James Baxter Bean, a Southern physician and dentist, invented the interdental splint. This device was used to treat hundreds of Confederate soldiers who had received gun shot-related facial and jaw injuries during the Civil War. Made of vulcanized India-rubber, the splint provided a dramatic breakthrough in the treatment of maxillofacial wounds. In an Atlanta, Georgia hospital, Dr. Bean utilized his invention by establishing the first ward devoted exclusively to the treatment of jaw fractures. He also invented an apparatus that manufactured and administered nitrous oxide. Additionally, Bean's groundwork in casting aluminum as a denture base material led to Taggart's later invention (in 1907) of the casting machine. After the Civil War, Dr. Bean became a highly successful dentist, practicing in Baltimore, Maryland. In the fall of 1870, at age 36, Bean, representing the Smithsonian Institution in Washington, D.C., traveled to Europe to gather geological specimens. A short time after arriving, Bean decided to climb Mont Blanc with ten other men. The entire group perished in a raging 8-day snow storm on the mountain peak. This tragedy, a compelling drama, is legendary in the annals of mountaineering history. After Dr. Bean's passing, his wife lost her sanity and subsequently died. Later, the death of the couple's only child, Chapin, sadly ended the family line. Although his life was cut short, Bean's contributions to dentistry have been significant and far-reaching. PMID- 12641170 TI - 19th century dentistry advertising trade cards. Gates of Pearl. PMID- 12641171 TI - William T.G. Morton and "The Great Moment". AB - The Great Moment, a Paramount movie released in 1944 about dentist William T.G. Morton's discovery of ether anesthesia a century earlier, was the odd-man-out among the movies made by the highly acclaimed director Preston Sturges in that period. It failed to attract large audiences and generally received only lukewarm reviews. Several biographies of Sturges have discussed the reasons for this anomaly; but only recently have drafts of the various versions of Sturges' scripts been published, plus additional background material about the film's production, revisions and editing. Using all this information, the author analyzes the movie and its history and asks what went wrong - and, more importantly, what went right. The general conclusion is that this little-known film has stood the test of time and is worthy of a revival among enthusiasts of dental history and a serious reassessment by movie critics in general. Despite some flaws in the final version, The Great Moment is in fact a remarkable medical biography, incorporating innovative flashback techniques and themes of inspiration and sacrifice mixed with some humor, while remaining reasonably true to historical facts surrounding dentistry's greatest triumph. PMID- 12641172 TI - Dentistry on currency. Tiradentes. PMID- 12641173 TI - The first dental college: emergence of dentistry as an autonomous profession. AB - Old as the ills of mankind, dentistry has been practiced as a specialty of surgery by physicians, surgeons, and artisans, at various times with titles such as barber surgeon, toothdrawer, operator for the teeth, and surgeon dentist. There being universal need for dental services, and lacking enforceable standards of practice and qualification, the profession was invaded by dubiously prepared pretenders at practice. The unconscionable consequence remained largely unremedied until realization of a philosophically sound basis of professional literature, organization, and education incident to the institution of the first dental college and the American system of dental education that elevated dentistry to the status of a recognized autonomous profession. PMID- 12641174 TI - Mechanisms underlying dependencies of performance on stimulus history in a two alternative forced-choice task. AB - In choice reaction time tasks, response times and error rates demonstrate differential dependencies on the identities of up to four stimuli preceding the current one. Although the general profile of reaction times and error rates, when plotted against the stimulus histories, may seem idiosyncratic, we show that it can result from simple underlying mechanisms that take account of the occurrence of stimulus repetitions and alternations. Employing a simple connectionist model of a two-alternative forced-choice task, we explored various combinations of repetition and alternation detection schemes in an attempt to account for empirical results from the literature and from our own studies. We found that certain combinations of the repetition and the alternation schemes provided good fits to the data, suggesting that simple mechanisms may serve to explain the complicated but highly reproducible higher order dependencies of task performance on stimulus history. PMID- 12641175 TI - A computational model of anterior cingulate function in speeded response tasks: effects of frequency, sequence, and conflict. AB - A growing body of evidence from functional neuroimaging and computational modeling studies indicates that the anterior cingulate cortex (ACC) detects the presence of response conflict and conveys this information to other brain regions, enabling subsequent adjustments in cognitive control. The present study examined previous empirical findings of increased ACC for low-frequency stimuli across three distinct speeded response tasks (two-alternative forced choice, go/no-go, and oddball). Simulations conducted in a neural network model incorporating sequential priming mechanisms (developed in Cho et al., 2002) confirmed that a computational measure of response conflict was higher on low frequency trials across all three tasks. In addition, the model captured detailed aspects of behavioral reaction time and accuracy data, predicted the dynamics of ACC activity related to trial sequence effects, and provided evidence for the functional role of conflict information in performance monitoring and optimization. The results indicate that the conflict-monitoring hypothesis, augmented by mechanisms for encoding stimulus history, can explain key phenomena associated with performance in sequential speeded response tasks. PMID- 12641176 TI - Selective hippocampal lesions disrupt a novel cue effect but fail to eliminate blocking in rabbit eyeblink conditioning. AB - The classical conditioning task of blocking involves the adding of a novel but redundant stimulus to a previously trained stimulus. Both blocking and novelty detection are thought to involve the hippocampus. Previously, Solomon (1977) found that nonselective aspiration lesions of the hippocampal region eliminated blocking in rabbit eyeblink conditioning. We tested the effects of selective ibotenic acid lesions of the hippocampus on blocking, as well as on novelty detection, when training is switched from a tone conditioned stimulus (CS) to a compound tone-light CS in eyeblink conditioning. Selective hippocampal lesions did not eliminate blocking but did lead to a facilitation of conditioned response (CR) acquisition to the tone and to the light, but not to the tone-light compound. Selective hippocampal lesions disrupted a CR decrement observed in sham surgical controls when transferred from tone training to tone-light training. It appears that although selective hippocampal lesions do not eliminate blocking in eyeblink conditioning, they do disrupt novelty detection and may facilitate learning to a previously blocked cue. PMID- 12641178 TI - Somatic markers, working memory, and decision making. AB - The somatic marker hypothesis formulated by Damasio (e.g., 1994; Damasio, Tranel, & Damasio, 1991) argues that affective reactions ordinarily guide and simplify decision making. Although originally intended to explain decision-making deficits in people with specific frontal lobe damage, the hypothesis also applies to decision-making problems in populations without brain injury. Subsequently, the gambling task was developed by Bechara (Bechara, Damasio, Damasio, & Anderson, 1994) as a diagnostic test of decision-making deficit in neurological populations. More recently, the gambling task has been used to explore implications of the somatic marker hypothesis, as well as to study suboptimal decision making in a variety of domains. We examined relations among gambling task decision making, working memory (WM) load, and somatic markers in a modified version of the gambling task. Increased WM load produced by secondary tasks led to poorer gambling performance. Declines in gambling performance were associated with the absence of the affective reactions that anticipate choice outcomes and guide future decision making. Our experiments provide evidence that WM processes contribute to the development of somatic markers. If WM functioning is taxed, somatic markers may not develop, and decision making may thereby suffer. PMID- 12641177 TI - Behavioral consequences of radiation exposure to simulated space radiation in the C57BL/6 mouse: open field, rotorod, and acoustic startle. AB - Two experiments were carried out to investigate the consequences of exposure to proton radiation, such as might occur for astronauts during space flight. C57BL/6 mice were exposed, either with or without 15-g/cm2 aluminum shielding, to 0-, 3-, or 4-Gy proton irradiation mimicking features of a solar particle event. Irradiation produced transient direct deficits in open-field exploratory behavior and acoustic startle habituation. Rotorod performance at 18 rpm was impaired by exposure to proton radiation and was impaired at 26 rpm, but only for mice irradiated with shielding and at the 4-Gy dose. Long-term (>2 weeks) indirect deficits in open-field activity appeared as a result of impaired experiential encoding immediately following exposure. A 2-week recovery prior to testing decreased most of the direct effects of exposure, with only rotorod performance at 26 rpm being impaired. These results suggest that the performance deficits may have been mediated by radiation damage to hippocampal, cerebellar, and possibly, forebrain dopaminergic function. PMID- 12641179 TI - The Union Carbide disaster in Bhopal: a review of health effects. AB - The authors have reviewed studies of human health effects that resulted from exposure to methyl isocyanate gas that leaked from the Union Carbide plant in Bhopal, India, in 1984. The studies were conducted during both the early and late recovery periods. Major organs exposed were the eyes, respiratory tract, and skin. Although mortality was initially high, it declined over time, but remained elevated among the most severely exposed population. Studies conducted during the early recovery period focused primarily on ocular and respiratory systems. Major findings included acute irritant effects on the eyes and respiratory tract. In follow-up studies, investigators observed persistent irritant effects, including ocular lesions and respiratory impairment. Studies conducted during the late recovery period focused on various systemic health endpoints. Significant neurological, reproductive, neurobehavioral, and psychological effects were also observed. Early and late recovery period studies suffered from several clinical and epidemiological limitations, including study design, bias, and exposure classification. The authors herein recommend long-term monitoring of the affected community and use of appropriate methods of investigation that include well designed cohort studies, case-control studies for rare conditions, characterization of personal exposure, and accident analysis to determine the possible components of the gas cloud. PMID- 12641180 TI - Investigation of health effects of hydrogen sulfide from a geothermal source. AB - Little is known about health effects from chronic exposure to hydrogen sulfide (H2S). The city of Rotorua, New Zealand, is exposed to H2S by virtue of its location over a geothermal field. In this study, the authors classified areas within Rotorua as high-, medium, or low-H2S exposure areas. Using 1993-1996 morbidity data, standardized incidence ratios were calculated for neurological, respiratory, and cardiovascular effects. Poisson regression analysis was used to confirm results. Results showed exposure-response trends, particularly for nervous system diseases, but also for respiratory and cardiovascular diseases. Data on confounders were limited to age, ethnicity, and gender. The H2S exposure assessment had limitations. Assumptions were that recent exposure represented long-term exposure and that an individual's entire exposure was received at home. The results of this study strengthen the suggestion that there are chronic health effects from H2S exposure. Further investigation is warranted. PMID- 12641181 TI - High prevalence of anti-glutamic acid decarboxylase (anti-GAD) antibodies in employees at a polychlorinated biphenyl production factory. AB - An increased prevalence of thyroid antibodies was seen in employees of a factory that formerly produced polychlorinated biphenyls (PCBs). In this study, the authors expand the evaluation of possible long-term PCB effects by comparing the prevalence of glutamic acid decarboxylase (anti-GAD) antibodies with the development of diabetes mellitus. The sera of 240 factory employees and 704 control subjects were analyzed. Anti-GAD antibody values exceeded 1.20 U/ml in all employees (40.4%), was 4 times higher (p < .001) than in all controls (10.5%), and were 5 times higher in employees aged 51-60 yr (53.2%) than in age matched controls (10.5%) (p < .001). Although the prevalence of diabetes could not be determined from this retrospective study, this is the first report of a possible relationship between xenobiotics and the prevalence of anti-GAD antibodies, and it supports the concept of an immunomodulatory effect of PCBs. However, such antibodies may be present decades before the development of clinical diabetes, and not all anti-GAD antibody-positive individuals become diabetic. Presently, it is unknown whether there is an increased prevalence of diabetes among the former factory employees. PMID- 12641183 TI - Smoking-induced squamous-cell cancer of the nose. AB - Squamous-cell cancer (SCC) isolated to the anterior nose is uncommon. Numerous industrial and environmental contaminants have been implicated as causative factors in nasal carcinoma. The association between nasal cancer and smoking was recognized in the 1980s, but results have not been consistent and no possible mechanisms have been suggested. The case histories of 11 patients with SCC limited to the anterior nasal cavity were reviewed. The authors found no relationship with exposures to chemicals previously implicated in nasal cancer. Nine patients with SCC were exposed to cigarette smoke (mean = 103 pack-years). Cigarette smoke may induce cancer directly, thus affecting the mucosa, or by inducing genetic alterations. The authors favor the former mechanism. PMID- 12641182 TI - Tobacco smoke and formation of N-(2-hydroxyethyl)valine in human hemoglobin. AB - Human exposure to ethylene oxide (EtO) occurs mainly through inhalation of polluted air in occupational workplaces and/or via tobacco smoke. A significant biochemical reaction of EtO converts the terminal valine of hemoglobin into N-(2 hydroxyethyl)valine (HOEtVal). In the present study, the extent of HOEtVal formation in 360 healthy adults who were not occupationally exposed to EtO was measured with a gas chromatograph/mass spectrometer in the electron-capture negative chemical ionization mode. This parameter was correlated with smoking habits and urinary cotinine concentration, exhibiting a positive relationship between HOEtVal and the number of cigarettes smoked (r2 = .4416). Urinary cotinine measurements also correlated with HOEtVal and the number of cigarettes smoked. This positive correlation between urinary cotinine and HOEtVal (r2 = .3893) provides a new perspective on the early stages of carcinogenic processes. PMID- 12641184 TI - Effect of environmental changes on oxidative deoxyribonucleic acid (DNA) damage in systemic lupus erythematosus. AB - In a study conducted in Japan, the authors used urinary 8-hydroxydeoxyguanosine (8-OHdG) to study the effects of high-intensity and low-intensity sunlight on oxidative damage to deoxyribonucleic acid (DNA) in patients who had systemic lupus erythematosus (SLE). During late May through early September (i.e., a period of high-intensity sunlight), the mean urinary 8-OHdG level in SLE patients was significantly higher than in controls (31.0 +/- 20.6 [standard deviation] ng/mg vs. 15.4 +/- 7.2 ng/mg, respectively [p < .05]). During late November through early March (i.e., low-intensity sunlight season), however, no significant differences were noted (15.4 +/- 5.5 ng/mg vs. 16.3 +/- 4.6 ng/mg, respectively). The mean urinary 8-OHdG level in SLE patients during the period of high-intensity sunlight was significantly higher than during the period of low intensity sunlight (21.3 +/- 20.6 ng/mg vs. 12.6 +/- 6.7 ng/mg, respectively; p < .01), although no such seasonal changes were observed among controls (16.2 +/- 8.0 ng/mg vs. 15.7 +/- 5.1 ng/mg, respectively). The effect of sunlight intensity (i.e., season) may require consideration when oxidative DNA damage occurs in individuals who have SLE. PMID- 12641185 TI - Symptomatology and etiology of multiple chemical sensitivities in the southeastern United States. AB - A questionnaire was administered to individuals who had reported a hypersensitivity to common chemical products in an earlier epidemiological study in the Atlanta, Georgia, metropolitan area. The questionnaire investigated the nature of the symptoms and factors that potentially initiated hypersensitivity and subsequently triggered reactions. Also examined were associated lifestyle modifications and the relationships of hypersensitivity with other illnesses. The authors found that a majority of hypersensitive individuals (52.2%) experienced either "severe" or "somewhat severe" symptoms. The most common triggers of symptoms were cleaning products (88.4%), tobacco smoke (82.6%), perfume (81.2%), pesticides (81.2%), and car exhaust (72.5%). Only 1.4% of the subjects had a prior history of emotional problems, whereas 37.7% developed such problems after the emergence of their hypersensitivity. Lifestyle modifications varied; 76.8% changed their household cleaning/personal hygiene products, 47.8% began using water and/or air filtration systems, and 13% found it necessary to change residence. Although hypersensitivity was more common in females than males, the condition affects individuals in all categories of race/ethnicity, age, household income, and educational level. PMID- 12641186 TI - Transient decrease of exhaled nitric oxide after acute exposure to passive smoke in healthy subjects. AB - Nitric oxide (NO) is produced and detected in the exhalate from the respiratory tract where it plays important regulatory functions. Exhaled nitric oxide (eNO) concentrations are reduced in active cigarette smokers between cigarettes and in nonsmoking subjects during short-term exposure to environmental tobacco smoke. In this study, the authors evaluated eNO before and after an acute exposure to environmental tobacco smoke in healthy, nonsmoking subjects (n = 12). Baseline eNO levels were measured by chemiluminescence at baseline (1 hr before exposure), shortly after the end of exposure, and 10 and 30 min after the end of exposure. Mean room air NO concentration increased from 3 ppb to 4 ppm (range, 560 ppb-8.5 ppm) during the exposure period. Carboxyhemoglobin levels were assessed before and after the exposure with spectrophotometry. All subjects had decreased eNO with exposure to environmental tobacco smoke (mean +/- standard error of the mean: 16.65 +/- 1.35 ppb to 13.86 +/- 1.33 ppb; p < .001). These concentrations remained significantly decreased at 10 min and recovered within 30 min. No modifications in airway resistance or increase in carboxyhemoglobin levels were observed. Exposure to environmental tobacco smoke transiently--but consistently- decreased eNO concentration in healthy, nonsmoking subjects, suggesting that second-hand smoke can directly affect NO in the airway environment. PMID- 12641187 TI - High prevalence of reversible airway obstruction in asbestos-exposed workers. AB - In this retrospective study of 97 male workers exposed to asbestos for 2-50 yr, data were collected on chest x-ray findings and lung function, including lung volumes, forced expiratory flow rates (i.e., forced expiratory volume in 1 sec [FEV1.0], forced expiratory flow measured between 25% and 75% of forced vital capacity [FEF(25-75%)]), airway resistance (R(aw)), carbon monoxide (CO) diffusing capacity, and the existence of airway obstruction reversible by a beta adrenergic agonist (RAO). The authors performed multiple-regression analyses to correlate the variations in lung function data with age, smoking habits, duration of asbestos exposure, and time since last exposure. Occupational activities of subjects that might induce specific lung abnormalities were also considered. No significant decrease was seen in lung volumes or CO-diffusing capacity; however, a decrease in FEV1.0 and an increase in R(aw) were measured in 65% of the subjects, and an isolated decrease in FEF(25-75%) occurred in only 18%. There was no difference in lung-function data between subjects who had chest x-ray signs of abnormalities (n = 59) and those who did not (n = 38). A significant relationship was found between the decrease in FEV1.0 and age; however, no correlation was noted between altered lung function and cigarette smoking, duration of asbestos exposure, or time since last exposure. RAO prevalence was higher (34%) than previously reported (9%) in subjects with chronic obstructive pulmonary disease (COPD) who were not exposed to asbestos or outdoor pollution. The RAO prevalence in asbestos-exposed workers was nearly the same as that measured in COPD subjects who lived downtown and who were exposed to outdoor pollution (36%). The high RAO prevalence in asbestos-exposed workers was found in 43% of subjects who were exposed only to asbestos, and in 33% of subjects who were also exposed to air pollution due to their occupational activities. It is hoped that the observations in this study will encourage practitioners to check for RAO in asbestos workers who use inhaled bronchodilators. PMID- 12641188 TI - Vanadium in ambient air: concentrations in lung tissue from autopsies of Mexico City residents in the 1960s and 1990s. AB - Vanadium concentrations in lung tissue were determined by atomic absorption spectrometry from autopsy specimens taken from residents of Mexico City during the 1960s and 1990s (20 males and 19 females, and 30 males and 18 females, respectively). Samples from the 1990s had significantly increased mean vanadium concentrations (mean +/- standard deviation: 1.36 +/- 0.08), compared with those from the 1960s (1.04 +/- 0.05). Concentrations were not correlated with gender, smoking habit, age, cause of death, or occupation. These findings suggest that vanadium in ambient air is increasing and it represents a potential health hazard for Mexico City residents. Air pollution monitoring efforts should include vanadium concentrations in suspended particles to follow-up the findings reported herein. Researchers need to acquire a better knowledge of the levels of airborne vanadium exposure at which risk to human health occurs. PMID- 12641190 TI - Association between petrochemical air pollution and adverse pregnancy outcomes in Taiwan. AB - The petrochemical industry is the main source of industrial air pollution in Taiwan. To date, little is known about the possible effects of such pollution on the human fetus. In this study, the authors investigated the relationship between term low birthweight and preterm delivery and residence in a petrochemical industrial municipality (PIM). The study areas included 16 PIMs (which were defined as municipalities in which the number of workers in the petroleum and petrochemical industry made up at least 2% of the municipality's total population) and 16 matched reference municipalities (RFMs). Among 39,750 1st parity singleton live births, the prevalences of preterm delivery in the PIMs and RFMs were 4.72% and 4.58%, respectively, and the prevalences of term low birthweight were 2.51% and 2.35%, respectively. When PIMs were compared with RFMs, and after controlling for possible confounders (including maternal age, marital status, maternal education, and gender of the baby), the adjusted odds ratios were 1.07 (95% confidence interval [CI] = 0.95, 1.22) for term low birthweight and 1.03 (95% CI = 0.94, 1.13) for preterm delivery, respectively. PMID- 12641189 TI - Assessment of physical education time and after-school outdoor time in elementary and middle school students in south Mexico City: the dilemma between physical fitness and the adverse health effects of outdoor pollutant exposure. AB - Strategies to promote lifelong physical activity among children are needed to stem the adverse health consequences of inactivity. However, the health effects in growing children of long-term exposure to a polluted atmosphere are of deep concern. The atmosphere of south Mexico City (SMC) is characterized by a complex mixture of air pollutants, including ozone, particulate matter, and aldehydes. Radiological evidence suggests that small-airway disease could be present in clinically healthy, tobacco unexposed SMC children. The aim of this study was to assess, by means of a self-reported questionnaire, the physical education class times, daily outdoor after-school exposure time, and tobacco exposure in students attending public elementary and middle schools in SMC. Additionally, the time each student spent viewing television was assessed, and the authors measured each student's weight and height to determine body mass index (BMI, weight in kg divided by height in m2). The survey included 1,159 students in grades 7-9. The authors identified 2 critical periods of outdoor exposure in SMC children that coincided with significant concentrations of both ozone and particulate matter with diameters less than 10 micrometers (PM10): during school time after 11:00 A.M. and in the after-school outdoor activity period, usually extending from 1:00 P.M. to 6:00 P.M. Thirty-two percent of elementary and 61% of middle school students have physical education classes after 11:00 A.M. Students in SMC spend an average of 19.6 hr/wk outdoors in the after-school period, during which time they are engaged in light to moderate physical activities. Half of the students are exposed to tobacco smoke at home, and 7% of middle school students smoke. On the basis of BMI, 60% of students were classified as undernourished, overweight, or obese. No correlations were found between BMI and time spent viewing TV, time outdoors (on weekdays and weekends), or exposure to environmental tobacco smoke. Children and adolescents in SMC are participating in physical activities that enhance multiple components of health-related fitness. However, their activities occur outdoors, where they are exposed to high concentrations of air pollutants throughout the year. The authors believe that SMC children and adolescents must be educated, through both the school and health systems, regarding ways to obtain the necessary exercise while protecting themselves from the high concentrations of pollutants. Individuals should instruct and encourage young people to be involved in lifetime fitness activities and to eat balanced diets, if the goal is to control health-care costs, reduce disease incidence, and improve the overall quality of life of the Mexico City population. PMID- 12641191 TI - Season modifies the relationship between bone and blood lead levels: the Normative Aging Study. AB - Bone serves as a repository for 75% and 90-95% of lead in children and adults, respectively. Bone lead mobilization heightens during times of increased bone turnover, such as pregnancy, lactation, hyperthyroidism, and the rapid growth of childhood. Blood lead levels show seasonal periodicity. Children demonstrate peak blood lead levels in mid-summer and a secondary peak in late winter. Pregnant women demonstrate the highest mean blood lead levels in winter (January-March) and the lowest in summer (July-September). This fluctuation in blood lead levels may be related to seasonal patterns of environmental exposures, but it may also be partially related to the increased mobilization of bone lead stores during the winter months. We performed bone lead measurements using a K-x-ray fluorescent instrument to determine micrograms of lead per gram of bone mineral (parts per million) in middle-aged and elderly men who participated in the Normative Aging Study. We obtained measurements of blood and bone lead during the high sun exposure months of May-August (summer; n = 290); the intermediate sun exposure months of March, April, September, and October (spring/fall; n = 283); and the low sun exposure months of November-February (winter; n = 191). Mean blood lead concentrations were 5.8 microg/dl, 6.1 microg/dl, and 6.6 microg/dl for the summer, spring/fall, and winter, respectively. Mean patella (trabecular bone) lead concentrations were 34.3 microg/gm, 29.7 microg/gm, and 29.0 microg/gm for the summer, spring/fall, and winter time periods, respectively. In multivariate regression models, adjusted for age, smoking, alcohol ingestion, and dietary intake of iron and vitamin C, the authors found a strong interaction between season and bone lead level--with bone lead levels exerting an almost 2-fold greater influence on blood levels during the winter months than the summer months. The authors concluded that elevated blood lead levels in winter may be related to increased mobilization of endogenous bone lead stores, potentially from decreased exposure to sunlight, lower levels of activated vitamin D, and enhanced bone resorption. PMID- 12641192 TI - Contaminant exposure among women of childbearing age who eat St. Lawrence River sport fish. AB - Little information is available concerning the level of consumption and degree of contaminant exposure for North American women of childbearing age who eat sport fish. The authors reanalyzed a 1995-1996 study of Montreal-area (Canada) sport fishers. The authors focused on women sport fishers of childbearing age and male sport fishers who had spouses of childbearing age. The primary research involved an on-site questionnaire about fish consumption, with follow-up assessment of sport fishers estimated to have either the highest or lowest levels of fish-based contaminant exposure. Among the 1,654 interviewees were 100 women less than 45 yr of age who had eaten sport fish for an average of 11 yr; 45% ate fish less than once a month. From the follow-up subsample of high- and low-level consumers, the authors identified 17 women less than 45 yr of age and 25 males whose spouses who were less than 45 yr of age and who consumed similar quantities of sport fish. Among this group of 42, the high-exposure women differed from the low-exposure women with respect to their yearly consumption of freshwater fish, blood mercury levels (median = 6.4 vs. 1.4 microgram/l), and plasma polychlorinated biphenyl congener 99 (median = 10.5 vs. 5.9 microgram/kg plasma lipids). Few Montreal-area women of childbearing age consume local sport fish frequently or for extended periods. However, among the small proportion that consumes sport fish frequently or for extended periods, blood mercury concentrations approach levels of concern for fetal protection. PMID- 12641193 TI - Effect of maternal bone lead on length and head circumference of newborns and 1 month-old infants. AB - The authors evaluated the effects that maternal bone lead stores have in anthropometry at birth in 223 mother-infant pairs. The participants were recruited between April and November 1994. Anthropometric data were collected within the first 12 hr following delivery. Maternal information was obtained 1 mo after delivery occurred. Bone lead burden was determined with in-vivo K-x-ray fluorescence of the tibia (cortical bone) and the patella (trabecular bone). The authors transformed anthropometric measurements to an ordinal 5-category scale, and the association of measurements with other factors was evaluated with ordinal logistic-regression models. Mean bone lead levels were 9.8 microgram/gm bone mineral and 14.4 microgram/gm bone mineral for the tibia and patella, respectively. Birth length of newborns decreased as tibia lead levels increased. Compared with women in the lower quintiles of the distribution of tibia lead, those in the upper quintile had a 79% increase in risk of having a lower birth length newborn (odds ratio = 1.79; 95% confidence interval = 1.10, 3.22). The authors adjusted by birth weight, and the effect was attenuated--but nonetheless significant. Patella lead was positively and significantly related to the risk of a low head circumference score; this score remained unaffected by inclusion of birth weight. The authors estimated the increased risk to be 1.02 per microgram lead/gm bone mineral (95% confidence interval = 1.01, 1.04 per microgram lead/gm bone mineral). Odds ratios did not vary substantially after the authors adjusted for birth weight and other important determinants of head circumference. PMID- 12641194 TI - Blood lead concentrations and pregnancy outcomes. AB - In this study, the authors related blood lead concentrations to Apgar scores, birth weight, gestational age, small-for-gestational age, and hypertension in pregnancy (HIP)/toxemia. Data and blood were collected 4 times during pregnancy from 705 women, aged 12-34 yr. Blood lead concentrations, measured by atomic absorption spectrophotometry, were related to reproductive outcomes, abstracted from medical records. Average blood lead concentrations were 1.2 microgram/dl (standard error = +/- 0.03). Maternal blood lead concentrations were related significantly to HIP/toxemia--before and after adjusting for age, calcium intake, and race/ethnicity (p < .03). Longitudinal regression analyses revealed that blood lead concentrations in women with HIP/toxemia changed by 0.02 microgram/dl for every 0.01 microgram/dl change in women without HIP/toxemia. Maternal blood lead concentration and its change were not significantly associated with other reproductive outcomes. Low levels of maternal blood lead concentrations were significantly associated with HIP/toxemia. PMID- 12641196 TI - Leptin and leptin receptors during malaria infection in mice. AB - Leptin, which is involved in a range of physiological processes, could be an important factor in the pathogenesis of malaria. We found that levels of leptin in serum and urine in Plasmodium berghei-infected mice increased progressively after infection, reaching a maximum value on day 6 post-infection. Serum values were approximately five-fold higher in infected mice than in non-infected controls. A similar relation was found for values of leptin in urine. Soluble leptin receptor levels also increased significantly in serum, more or less in line with the leptin increase. Our work represents the first report of visibly augmented leptin and soluble leptin receptor secretion in malarial infection. PMID- 12641195 TI - Dietary intakes and plasma organochlorine contaminant levels among Great Lakes fish eaters. AB - Nutritional intakes and contaminant burdens should be assessed jointly in individuals who are at high risk of environmental exposures to contaminants through food. In this study, the authors used shore surveys and community contacts to recruit 91 individuals who frequently consumed Great Lakes fish. These individuals provided dietary intake information and fasting blood samples for lipid and contaminant analyses. Participants ate an annual median of 88 meals of Great Lakes fish. Asian-Canadians consumed more total fish meals (i.e., Great Lakes, non-Great Lakes, and other) (medians = 213.0 females, 223.0 males) than Euro-Canadians (medians = 131.0 females, 137.5 males). The higher total fish consumption by Asian-Canadians was associated with a lower percentage of energy derived from fat, higher protein and iron intakes, and higher plasma concentrations of omega-3 essential fatty acids (e.g., median docosahexaenoic acid levels [microgram/l] in Asian-Canadian females = 5.48, males = 4.38; in Euro Canadian females = 2.93, males = 2.27). Plasma organochlorine contaminant lipid weight concentrations varied by country of origin and by gender (e.g., median total polychlorinated biphenyls [microgram/kg] in Asian-Canadian females = 490.6, males = 729.0; in Euro-Canadian females = 339.6, males = 355.5). Age was the most consistent predictor (+ve) of contaminant concentrations, followed by years spent in Canada (for Asian-Canadians). Associations with sport fish consumption variables were less consistent than for the aforementioned predictors. Given both the health benefits and potential risks of fish consumption, policies that address diverse ethnocultural groups should support continued consumption of sport fish, but from less-contaminated sources than are currently used. PMID- 12641198 TI - Light and scanning electron microscopy of Myxobolus porofilus sp. n. (Myxosporea: Myxobolidae) infecting the visceral cavity of Prochilodus lineatus (Pisces: Characiformes: Prochilodontidae) cultivated in Brazil. AB - Myxobolus porofilus sp. n. is described infecting the visceral cavity of Prochilodus lineatus (Valenciennes, 1836) cultivated in Sao Paulo State, Brazil. The plasmodial form of the parasite is 3-5 mm in length and appeared compressed between the wall of the visceral cavity and the pyloric caecum, reposing on this organ. The spores are small (length 5.7 +/- 0.3 microm, width 4.8 +/- 0.2 microm; mean +/- SD) and round to elliptical in frontal view. The valve surfaces are smooth and have sutural folds. The polar capsules are ovoid, small (length 1.6 +/ 0.1 microm, width 1.1 +/- 0.1 microm) and equal in size. The polar filaments have three turns aligned perpendicularly to the longitudinal axis of the capsule. A conspicuous polar filament pore is arranged at the anterior end of the spore. The only reaction observed upon histological analysis was the presence of a capsule of connective tissue surrounding the plasmodia. This is the first report of a myxosporean parasite in the Prochilodontidae. PMID- 12641197 TI - Myxobolus lentisuturalis sp. n. (Myxozoa: Myxobolidae), a new muscle-infecting species from the Prussian carp, Carassius gibelio from China. AB - A new highly pathogenic muscle-infecting species of the genus Myxobolus Butschli, 1882 is described from the Prussian carp, Carassius gibelio (Bloch, 1782) using spore morphology and SSU rDNA sequence data. Phylogenetic analyses elucidated relationship of the newly described Myxobolus lentisuturalis to other Myxobolus species and supported its position of an independent species. PMID- 12641199 TI - Infection prevalence, seasonality and host specificity of actinosporean types (Myxozoa) in an Atlantic salmon fish farm located in Northern Scotland. AB - A total of 28,387 oligochaetes belonging to the families Tubificidae, Lumbriculidae, Naididae and Enchytraeidae were examined at regular intervals for actinosporean infections from October 1996 to August 1998 from a freshwater salmon farm in Northern Scotland. A total of 21 types of actinosporeans belonging to seven collective groups synactinomyxon (three types), aurantiactinomyxon (four types), echinactinomyxon (five types), raabeia (six types), triactinomyxon (one type), neoactinomyxum (one type) and siedleckiella (one type) were found. Synactinomyxon type 1, echinactinomyxon type 1 and raabeia type 4 were most abundant. The overall infection prevalence of oligochaetes was 2.9%. Aurantiactinomyxon, synactinomyxon and neoactinomyxum were most common in summer and autumn. Raabeia was most common in spring and summer and echinactinomyxon in winter and spring. Siedleckiella was found only in spring and triactinomyxon in all seasons except winter. A positive relationship between water temperature and the number of actinosporean types released was observed. Most actinosporean types were found in only one host species. PMID- 12641200 TI - Trickle and single infection with Discocotyle sagittata (Monogenea: Polyopisthocotylea): effect of exposure mode on parasite abundance and development. AB - Experimental infection of rainbow trout Oncorhynchus mykiss (Walbaum) with the monogenean Discocotyle sagittata (Leuckart, 1842) allowed comparison between trickle and single exposure, two infection modes demonstrated to occur in the wild. Both types of infection resulted in mean larval attachment success around 50%, which was significantly dependent on dose of infective larvae used (P < 0.0001), but was not affected by mode of infection (P = 0.244). Worms recovered from fish exposed to the same number of oncomiracidia but different mode of infection differed in their rate of development. The developmental stage attained by parasites was significantly affected by number of infective larvae used (P = 0.005), and by the interaction between dose and mode of infection (P = 0.026), suggesting competition among attached larvae. Statistical analysis demonstrated that in the early stages of infestation, worm distribution over the gill arches can be explained by the relative amount of water flowing over them. One, two and three months post-infection parasite numbers were comparable (P = 0.805), but their observed distribution gradually decreased in gill arches III and IV and increased in gill arch I, suggesting that parasites migrate after initial attachment. These results reproduce phenomena observed in the field, indicating that the experimental infection system could be employed to study infection dynamics and host-parasite interactions under controlled conditions. PMID- 12641201 TI - Observations on the phylogeny of Opisthadena Linton, 1910 and related genera (Hemiuridae: Opisthadeninae) from Australian and French Polynesian waters. AB - Mitrostoma nototheniae Manter, 1954 is redescribed from Aplodactylus arctidens Richardson, off northern Tasmania. Opisthadena dimidia Linton, 1910 is reported from Kyphosus bigibbus Lacepede, Ningaloo, Western Australia, K. cinerascens (Forsskal), off Heron Island and Ningaloo, Western Australia, Kyphosus cornelii (Whitley), off Kalbarri, Western Australia, K. sydneyanus Gunther, off Fremantle, Western Australia, K. sydneyanus ?, Ningaloo, Western Australia and K. vaigiensis (Quoy et Gaimard), off Heron and Lizard Islands, Queensland and Moorea, French Polynesia: measurements and an illustration are given. Neopisthadena habei Machida, 1980 is reported from K. sydneyanus off Fremantle, Western Australia: measurements and an illustration are given. Data derived from these specimens are used to recode a data-matrix developed by Leon-Regagnon et al. (1996), and the resultant tree produced is almost congruent with that of these authors. Our data indicate that this group of parasites is associated mostly with herbivorous hosts and that O. dimidia, while geographically widespread, is stenoxenic to the genus Kyphosus. PMID- 12641202 TI - Ultrastructure of the forebody and foregut tegument and eccrine gland cells of Crepidostomum metoecus (Trematoda: Digenea: Allocreadiidae). AB - The forebody and foregut of Crepidostomum metoecus Braun, 1900 are invested with a tegument bearing regularly arranged surface tubercles comparable with the aspidogastrean surface structures. The tegument of the ventrolateral lobes and of the prepharynx is penetrated by ducts of eccrine gland cells. The frontal and prepharyngeal gland cells, localised in the parenchyma, discharge electron-dense granules. Their ducts are lined by peripheral microtubules and fixed to the tegument plasmalemma by a septate junction. The functional roles of these glands are discussed. PMID- 12641203 TI - Anthobothrium altavelae sp. n. (Cestoda: Tetraphyllidea) from the spiny butterfly ray Gymnura altavela (Elasmobranchii: gymnuridae) in Tunisia. AB - Seven Gymnura altavela (Linnaeus, 1758) (Elasmobranchii, Myliobatiformes) caught off the Tunisian coast were examined for endoparasites during a three-year period (1995-1998). A phyllobothriid cestode new to science was found in the spiral intestine of all host specimens. The presence of a tetrabothridiate scolex, bothridia lacking an apical sucker, laciniate strobila, and possession of postvaginal testes are sufficient to place this species in the genus Anthobothrium Van Beneden, 1850. In this genus, we consider that only two species, both previously reported from carcharhiniform sharks, are valid: Anthobothrium cornucopia Van Beneden, 1850 and A. laciniatum Linton, 1890. Anthobothrium altavelae sp. n. can be distinguished from its congeners by its length, strobila morphology, and the number of testes. The taxonomic position of the species attributed to Anthobothrium parasitic in gymnurids is discussed. PMID- 12641204 TI - Moravecnema segonzaci gen. et sp. n. (Nematoda: Cystidicolidae) from Pachycara thermophilum (Zoarcidae), a deep-sea hydrothermal vent fish from the Mid-Atlantic Ridge. AB - A new cystidicolid nematode, Moravecnema segonzaci gen. et sp. n. is described from the intestine of the teleost fish Pachycara thermophilum Geistdoerfer (Zoarcidae) from the hydrothermal sites Logatchev and Snake Pit-Moose of the Mid Atlantic Ridge, at depths of 3,008; 3,492, and 3,510 m. The new genus Moravecnema is characterised by a dorsoventrally elongated oral opening, rudimentary pseudolabia, and four pairs of precloacal and six pairs of postcloacal caudal papillae in the male. The species has two spicules of unequal length, about 330 and 80 microm long. Males are about 5 mm and females about 5-10 mm long. Eggs have long thin polar filaments. This is the first species of parasitic nematode described from a fish endemic to hydrothermal deep-sea vents. PMID- 12641205 TI - Three new species of the ectoparasitic mites of the genus Syringophiloidus Kethley, 1970 (Acari: Syringophilidae) from passeriform birds from Slovakia. AB - Three new quill mite species of the genus Syringophiloidus Kethley, 1970 (Acari: Syringophilidae) from Slovakia are described and figured: S. bombycillae sp. n. from the waxwing Bombycilla garrulus (L.) (Passeriformes: Bombycillidae), S. schoeniclus sp. n. from the reed bunting Emberiza schoeniclus L. (Passcriformes: Emberizidae) and S. montanus sp. n. from the tree sparrow Passer monlanus (L.) (Passeriformes: Ploceidae). A key to all known species of the genus Syringophiloidus is given. PMID- 12641206 TI - Nymphal diapause and its photoperiodic control in the tick Ixodes scapularis (Acari: Ixodidae). AB - It is shown experimentally that the option between developmental diapause and non diapause development in nymphs of Ixodes scapularis Say, 1821 (Middle Atlantic population) is determined by photoperiodic conditions according to a two-step photoperiodic reaction of short-day long-day type. Diapause arrest of development is induced by an impact of either long day upon unfed nymphs, or short day upon engorged nymphs, while non-diapause development completed in 2-2.5 months at 20 degrees C needs the change from short-day to long-day conditions. Some ecophysiological aspects of mechanisms controlling seasonal development of ticks belonging to Ixodes ricinus complex are discussed. PMID- 12641207 TI - Screening of the T- and B-cell antigenicity in neonatal calves of the His-tagged Cryptosporidium parvum antigens CP15, CP15/60, P23 and TRAP-C1. PMID- 12641208 TI - Potential significance of transovarial transmission in the circulation of tick borne encephalitis virus. PMID- 12641210 TI - Endoplasmic reticulum-associated protein degradation. AB - Proteins that fail to fold properly as well as constitutive or regulated short lived proteins of the endoplasmatic reticulum (ER) are subjected to proteolysis by cytosolic 26 S proteasomes. This process, termed ER-associated protein degradation (ERAD), has also been implicated in the generation of some important human disorders, for example, cystic fibrosis. To become accessible to the proteasome, ERAD substrates must first be retrogradely transported from the ER into the cytosol, in a process termed dislocation. Surprisingly, protein dislocation from the ER seems to require at least some components that also mediate import into this compartment. Moreover, polyubiquitination of ERAD substrates at the ER membrane as well as the cytoplasmic Cdc48p/Npl4p/Ufd1p complex were shown to contribute to this export reaction. In this article we will summarize our current knowledge on ERAD and discuss the possible function of certain components involved in this process. PMID- 12641209 TI - Thymic nurse cells: a microenvironment for thymocyte development and selection. AB - Thymic nurse cells (TNCs) represent a unique microenvironment in the thymus for MHC restriction and T cell repertoire selection composed of a cortical epithelial cell surrounding 20-200 immature thymocytes. TNCs have been isolated from many classes of animals from fish to humans. Studies performed using TNC lines showed that TNCs bind viable alphabetaTCRlow CD4(+)CD8(+)CD69(-) thymocytes. A subset of the bound cells is internalized, proliferates within the TNC, and matures to the alphabetaTCRhigh CD4(+)CD8(+)CD69(+) stage, indicative of positive selection. A subset of the internalized population is released while cells that remain internalized undergo apoptosis and are degraded by lysosomes within the TNC. A TNC-specific monoclonal antibody added to fetal thymic organ cultures resulted in an 80% reduction in the number of thymocytes recovered, with a block at the double positive stage of development. Together these data suggest a critical role for TNC internalization in thymocyte selection as well as the removal and degradation of negatively selected thymocytes. Recent studies have shown that in addition to thymocytes, peripheral circulating macrophages are also found within the TNC complex and can present antigens to the developing thymocytes. These circulating macrophages could provide a source of self-antigens used to ensure a self-tolerant mature T cell repertoire. A reduction in TNC numbers is associated with a variety of autoimmune diseases including thyroiditis and systemic lupus erythematosis. PMID- 12641211 TI - Functional complexity of intermediate filament cytoskeletons: from structure to assembly to gene ablation. AB - The cell biology of intermediate filament (IF) proteins and their filaments is complicated by the fact that the members of the gene family, which in humans amount to at least 65, are differentially expressed in very complex patterns during embryonic development. Thus, different tissues and cells express entirely different sets and amounts of IF proteins, the only exception being the nuclear B type lamins, which are found in every cell. Moreover, in the course of evolution the individual members of this family have, within one species, diverged so much from each other with regard to sequence and thus molecular properties that it is hard to envision a unifying kind of function for them. The known epidermolytic diseases, caused by single point mutations in keratins, have been used as an argument for a role of IFs in mechanical "stress resistance," something one would not have easily ascribed to the beaded chain filaments, a special type of IF in the eye lens, or to nuclear lamins. Therefore, the power of plastic dish cell biology may be limited in revealing functional clues for these structural elements, and it may therefore be of interest to go to the extreme ends of the life sciences, i.e., from the molecular properties of individual molecules including their structure at the atomic level to targeted inactivation of their genes in living animals, mouse, and worm to define their role more precisely in metazoan cell physiology. PMID- 12641212 TI - Morphodynamics of the follicular-luteal complex during early ovarian development and reproductive life. AB - Female reproductive activity depends upon cyclic morphofunctional changes of the ovarian tissue during the female's fertile period, but the primum movens of an active gonadal rearrangement can be found from early phases of embryo development. To offer a basic account of the main steps of ovarian dynamics, we review the morphofunctional behavior of the follicular-luteal complex in an integrated study using light microscopy and transmission and scanning electron microscopy as well as through the use of numerous drawings. Particular emphasis is given to some reproductive aspects including (1) germ-somatic cell relationships and onset of folliculogenesis during early gonadal development; (2) follicular development and oocyte-follicle cell associations through adult folliculogenesis, finally leading to ovulation; (3) morphodynamics of corpus luteum formation, development, and regression, and (4) degenerative processes involving germ and somatic cells. The results reported, many of which originated in our laboratory, arise from some experiments on laboratory mammals but mostly from a large selection of human specimens. The data obtained are integrated and correlated with classic reports as well as with current views. Crucial biochemical, histophysiological, and clinical aspects are also emphasized. PMID- 12641222 TI - Development of a multiple-class high-resolution gas chromatographic relative retention time model for halogenated environmental contaminants. AB - A predictive model for the relative gas chromatographic retention times (GC-RRTs) of the following nine classes of halogenated environmental contaminants was developed: polybrominated diphenyl ethers (PBDEs); polychlorinated diphenyl ethers (PCDEs); polychlorinated biphenyls (PCBs); polychlorinated naphthalenes (PCNs); polychlorinated dibenzo-p-dioxins (PCDDs); polychlorinated dibenzofurans (PCDFs); polybrominated dibenzo-p-dioxins (PBDDs); polybrominated dibenzofurans (PBDFs); and organochlorine pesticides. MOPAC calculated physicochemical properties and structural descriptors in the model include molecular weight, square root of the number of halogen substituents, ionization potential, dipole moment, and the number of ortho, meta, and para halogen substituents. Using these variables, individual models for each of the contaminant classes were combined into a multiple class model incorporating the GC-RRTs of the 375 compounds of interest. The individual and multiclass GC-RRT models had acceptable fits between observed and predicted GC-RRTs (r2 = 0.9741-0.9990 for PBDEs, PCDEs, PCBs, PCNs, PCDD/Fs, and PBDD/Fs; r2 = 0.9250 for pesticides; and r2 = 0.9631 for the multiclass model) over a wide range of retention times and molecular structures. The combined model was tested on known GC-RRTs of hydroxylated PCBs and chlorinated phenoxyphenols and provided satisfactory results, demonstrating the strength of the model in predicting GC-RRT windows for contaminant classes not used in constructing the model. Such models will be useful in predicting the GC retention characteristics of novel environmental contaminants and their degradation products, for which analytical standards may not be available. PMID- 12641221 TI - Use of artificial neural networks for the accurate prediction of peptide liquid chromatography elution times in proteome analyses. AB - The use of artificial neural networks (ANNs) is described for predicting the reversed-phase liquid chromatography retention times of peptides enzymatically digested from proteome-wide proteins. To enable the accurate comparison of the numerous LC/MS data sets, a genetic algorithm was developed to normalize the peptide retention data into a range (from 0 to 1), improving the peptide elution time reproducibility to approximately 1%. The network developed in this study was based on amino acid residue composition and consists of 20 input nodes, 2 hidden nodes, and 1 output node. A data set of approximately 7000 confidently identified peptides from the microorganism Deinococcus radiodurans was used for the training of the ANN. The ANN was then used to predict the elution times for another set of 5200 peptides tentatively identified by MS/MS from a different microorganism (Shewanella oneidensis). The model was found to predict the elution times of peptides with up to 54 amino acid residues (the longest peptide identified after tryptic digestion of S. oneidensis) with an average accuracy of approximately 3%. This predictive capability was then used to distinguish with high confidence isobar peptides otherwise indistinguishable by accurate mass measurements as well as to uncover peptide misidentifications. Thus, integration of ANN peptide elution time prediction in the proteomic research will increase both the number of protein identifications and their confidence. PMID- 12641223 TI - Methodological refinements in the determination of 146 polychlorinated biphenyls, including non-ortho- and mono-ortho-substituted PCBs, and 26 organochlorine pesticides as demonstrated in heron eggs. AB - A method for the determination of 146 polychlorinated biphenyls (PCBs), including four non-ortho- and eight mono-ortho-substituted congeners, and 26 chlorinated pesticides is described. The method consists of ultrasonic extraction, Florisil cleanup, HPLC fractionation over porous graphitic carbon (PGC), and final determination with GC/ECD, GC/MS, or both. Two PCB congeners (PCB 30 and PCB 161) and two polybromobiphenyls (2,4',5-tribromobiphenyl and 3,3',4,4' tetrabromobiphenyl) were used as surrogate standards to evaluate the analytical efficiency. Four PCB congeners, PCB 14 and PCB 159 for the first fraction, PCB 61 for the second fraction, and PCB 204 for the third fraction, were used as internal standards to monitor the GC performance. The retention behavior of PCBs and pesticides on the PGC column are discussed. The method was found to be reproducible, effective, and reliable under the operational conditions proposed and was applied successfully to the analysis of individual PCBs and chlorinated pesticides in black-crowned night heron (Nycticorax nycticorax) egg samples. PMID- 12641224 TI - Membrane-based nanoscale proteolytic reactor enabling protein digestion, peptide separation, and protein identification using mass spectrometry. AB - A miniaturized trypsin membrane reactor housed inside a commonly used capillary fitting is developed and demonstrated for enabling rapid and sensitive protein identification by on-line proteolytic digestion and analysis of protein digests using nano-ESI-MS and MALDI-MS. The design and assembly of the capillary fitting based trypsin membrane reactor are straightforward and highly robust, without the need for expensive fabrication technology and procedures. The resultant protein digests can also be further concentrated and resolved using capillary reversed phase liquid chromatography or transient capillary isotachophoresis/zone electrophoresis prior to the mass spectrometric analysis in an integrated platform. By comparing these results with the results obtained from our previous studies using plastic microfluidics (Gao et al., Anal. Chem. 2001, 73, 2648 2655), significant reduction in dead volume and sample consumption can be achieved using this newly developed tryptic digestion station. This nanoscale reaction system enables rapid proteolytic digestion in seconds instead of hours for a protein concentration of less than 10(-8) M, consumes very little sample (< or = 5 fmol), and offers capillary interfaces with various separation and mass spectrometry techniques. The ultrafast enzymatic turnover for attaining complete peptide coverage in protein identification is contributed by the highly porous structure of the membrane media, providing excessive trypsin loading while eliminating the constraints of diffusion-limited reaction kinetics. PMID- 12641225 TI - Measurement of attogram quantities of 231Pa in dissolved and particulate fractions of seawater by isotope dilution thermal ionization mass spectroscopy. AB - A technique has been developed to quantify ultratrace 231Pa (50-2000 ag; 1 ag = 10(-18) g) concentrations in seawater using isotope-dilution thermal ionization mass spectrometry (TIMS). The method is a modification of a process developed by Pickett et al. (Pickett, D. A.; Murrell, M. T.; Williams, R. W. Anal. Chem. 1994, 66, 1044-1049) and extends the technique to very low levels of protactinium. The procedural blank is 16 +/- 15 ag (2sigma), and the ionization efficiency (ions generated/atom loaded) approaches 0.5%. Measurement time is <1 h. The amount of 231Pa needed to produce 231Pa data with an uncertainty of +/-4-12% is 100-1000 ag (approximately 3 x 10(5) to 3 x 10(6) atoms). Replicate measurements made on known standards and seawater samples demonstrate that the analytical precision approximates that expected from counting statistics and that, based on detection limits of 38 and 49 ag, protactinium can be detected in a minimum sample size of surface seawater of approximately 2 L for suspended particulate matter and <0.1 L for filtered (<0.4 microm) seawater, respectively. The concentration of 231Pa (tens of attograms per liter) can be determined with an uncertainty of +/-5-10% (2sigma) for suspended particulate matter filtered from 5 to 10 L of seawater. For the dissolved fraction, 0.5-1 L of seawater yields 231Pa measurements with a precision of 1-10%. Sample size requirements are orders of magnitude less than traditional decay-counting techniques and significantly less than previously reported ICP-MS techniques. Our technique can also be applied to other environmental samples, including cave waters, rivers, and igneous rocks. PMID- 12641226 TI - Broadband coupling into a single-mode, electroactive integrated optical waveguide for spectroelectrochemical analysis of surface-confined redox couples. AB - A single-mode, electroactive waveguiding platform capable of measuring spectroelectrochemical responses of surface-adsorbed redox-active molecules over a broad spectral bandwidth has been created. This new planar waveguide spectrometer is a combination of the previously developed electroactive integrated optical waveguide (EA-IOW; Dunphy, D. R.; Mendes, S. B.; Saavedra, S. S.; Armstrong, N. R. Anal. Chem. 1997, 69, 3086-3094) with a recently reported simplified approach to broadband coupling (Bradshaw, J. T.; Mendes, S. B.; Saavedra, S. S. Anal. Chem. 2002, 74, 1751-1759). With the use of a commercially available prism as an incoupling element, the EA-IOW can now guide visible light from at least 500 to 700 nm, improving upon its previously demonstrated monochromatic nature. Coupling profiles of various laser lines along with transmission spectra of narrow band-pass filters at various potentials are used to demonstrate the optical characteristics of this broadband EA-IOW and to compare its response to that of a conventional transmission instrument. Assessment of spectral resolution, performed by measuring the fwhm of various laser lines, ranges from 0.6 to 0.8. To demonstrate the capabilities of this technology, we show the acquisition of absorbance spectra of two different adsorbates, cytochrome c and ferrocenedicarboxylic acid, as a function of applied potential. Subtleties in the redox chemistries of adsorbed molecules, which were difficult to monitor with a monochromatic waveguide, are readily apparent when using the broadband coupling scheme. PMID- 12641227 TI - Scanning tunneling microscopy with chemically modified gold tips: in situ reestablishment of chemical contrast. AB - A method was developed for the reestablishment of chemical contrast in STM images obtained with chemically modified gold tips. Such tips display selective chemical contrast, which allows the selective imaging of specific species on the sample surface. Chemically modified STM tips can be fabricated by forming a self assembled monolayer (SAM) on an electrochemically etched gold tip. One difficulty with this method thus far has been the relatively short lifetime of SAM-treated tips. The method described here utilizes the brief application of a high bias voltage between the sample and the tip to cause SAM molecules to reoccupy the tip apex, thereby allowing the tips to display selective chemical contrast in imaging. These treatments consist of applying a +1.9-V sample bias for 0.5-10 min under tunneling conditions. The usable lifetime of SAM-modified tips could be increased by more than 2 orders of magnitude, from hours to at least a month, dramatically increasing the efficiency of using SAM-modified gold tips. SAM molecules can also be removed from the tip apex by application of a negative sample bias (-2.0 V for 0.5-10 min) making it possible to alternate between conventional STM images and STM images with chemically enhanced contrasts. PMID- 12641229 TI - Electrochemiluminescence of tris(8-hydroxyquinoline-5-sulfonic acid)aluminum(III) in aqueous solution. AB - The electrochemiluminescence (ECL) of tris(8-hydroxyquinoline-5-sulfonic acid)aluminum(III) in aqueous solution is reported. ECL is generated by complexing aluminum ions with the chelating agent 8-hydroxyquinoline-5-sulfonic acid (HQS) to form Al(HQS)3, followed by oxidation in the presence of tri-n propylamine (TPrA). The ECL intensity peaks a potential corresponding to oxidation of both TPrA and Al(HQS)3, and the ECL emission spectrum (lambda(max) = 499 nm) matches the photoluminescence emission spectrum, indicating that the emission is from a Al(HQS)3* excited state. ECL efficiencies (phi(ecl), photons generated per redox event) of 0.002 using Ru(bpy)3(2+) (phi(ecl) = 1) as relative standard. Conditions for ECL emission were optimized and used to generate a calibration curve that was linear over the 7 x 10(-6)-4 x 10(-4) M (5-281 mg/L (ppm)) range with a theoretical limit of detection of 1 ppm. The ECL of several metal ions other than aluminum with HQS and effects on Al(HQS)3 ECL were also examined. PMID- 12641228 TI - Ion sensing and inhibition studies using the transmembrane ion channel peptide gramicidin A entrapped in sol-gel-derived silica. AB - The development of new, targeted drugs relies heavily on innovative technologies that allow for high-throughput screening of drug libraries against biologically relevant targets, particularly membrane-associated receptors. Therefore, immobilization of natural receptors is of the utmost importance to allow for screening of small molecule libraries. Herein, we describe the immobilization of liposomes containing the transmembrane peptide ion-channel gramicidin A into sol gel-derived silicate materials. Steady-state fluorescence measurements of the intrinsic tryptophan residues of reconstituted gramicidin A in phospholipid vesicles consisting of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) were obtained in solution and following entrapment in diglyceryl silane (DGS)-derived silicate to examine the effects of entrapment on the conformation of the ion channel. Only minor deviations were observed in the fluorescence properties of gramicidin following entrapment in DGS-derived silicate. DOPC vesicles containing a 50 microM internal solution of the potential sensitive fluorescent dye safranine O were used to study ion flux through the membrane ion channel. The dependence of ion flux on both ion concentration and amount of gramicidin embedded in the membrane were examined before and after entrapment in sol-gel derived silicate. It was found that ion channel activity upon entrapment in DGS derived silicate mirrored very closely that observed in solution. Moreover, the ability to inhibit ion flux through gramicidin A due to blockage by calcium ions was retained after the immobilization procedure. The implications for development of drug-screening and -sensing platforms are discussed. PMID- 12641230 TI - Model-based optimal design of polymer-coated chemical sensors. AB - A model-based methodology for optimal design of polymer-coated chemical sensors is developed and is illustrated for the example of infrared evanescent field chemical sensors. The methodology is based on rigorous and computationally efficient modeling of combined fluid mechanics and mass transfer, including transport of multiple analytes. A simple algebraic equation for the optimal size of the sensor flow cell is developed to guide sensor design and validated by extensive CFD simulations. Based upon these calculations, optimized geometries of the sensor flow cell are proposed to further improve the response time of chemical sensors. PMID- 12641231 TI - Electrochemical protein chip with arrayed immunosensors with antibodies immobilized in a plasma-polymerized film. AB - An electrochemical protein chip was microfabricated. A thin-film three-electrode system, including an array of 36 platinum working electrodes, a set of thin-film Ag/AgCl electrodes, and platinum auxiliary electrodes, was integrated on a glass substrate. Capture antibodies were immobilized in a 4.5-nm-thick double layer of a hexamethyldisiloxane plasma-polymerized film. Because of their highly cross linked network structure, the capture antibodies could be firmly immobilized. No nonspecific adsorption was observed during a series of procedures to detect target proteins, and electrochemical cross talk between neighboring sites was negligible. The sandwich immunoassay was conducted on a single chip using model proteins, alpha-1-fetoprotein and beta2-microglobulin. A distinct current increase following the oxidation of hydrogen peroxide produced by the enzymatic reaction of glucose oxidase was observed, which indicates that the capture proteins could actually bind the target proteins. Two kinds of protein were detected independently on multiple sites with respective capture antibodies. PMID- 12641232 TI - Characterization of an EDTA bonded conducting polymer modified electrode: its application for the simultaneous determination of heavy metal ions. AB - An EDTA bonded conducting polymer modified electrode (EDTA-CPME) was fabricated by polymerization of 3',4'-diamino-2,2';5',2''-terthiophene monomer on a GCE, followed by the reaction with EDTA in the presence of catalyst. The surface of the resulting modified electrode was characterized with EQCM, ESCA, SEM, Auger electron spectroscopy, scanning Auger microscopy, and electrochemical methods. The amounts of polymer and EDTA attached on the polymer film were determined. Simple immersing of the EDTA-CPME into a sample solution led to the chemical deposition through the complexation with Pb2+, Cu2+, and Hg2+ ions, simultaniously. Various experimental parameters that affect the simultaneous analysis of the metal ions, e.g., EDTA amount, pH, deposition time, and deposition temperature, were optimized. Calibration plots for the EDTA-CPME with square wave voltammetry were obtained in the concentration range between 5.0 x 10(-10) and 1.0 x 10(-7) M for Cu(II) and between 7.5 x 10(-10) and 1.0 x 10(-7) M for Pb(II) and Hg(II). The detection limits for Pb(II), Cu(II), and Hg(II) ions were determined to be about 6.0 x 10(-10), 2.0 x 10(-10), and 5.0 x 10(-10) M, respectively. Interference effects from other metal ions were studied at various pHs and it was found that there was little or no effect on the simultaneous determination. The stability of the EDTA-CPME was remarkably improved by coating the surface with the Nafion film, and the electrode can be used for more than one month. Analytical availability of the EDTA-CPME was demonstrated by the application for the certified standard urine reference material and tap water. PMID- 12641234 TI - Immobilized particle arrays: coalescence of planar- and suspension-array technologies. AB - Combining positive attributes of planar arrays and suspension arrays, immobilized particle arrays offer a new format in which immobilized submicrometer particles are arrayed on hydrogel-coated slides, providing 100+ assay replicates within each spot. This research describes how to prepare immobilized protein arrays and how to assay the binding of labeled target molecules to the arrayed capture probes. The assay system exhibits an intrinsic dynamic range of two to three decades, with coefficients of variation from 5 to 10%. For antibody-antigen binding, target capture appears to be reaction rate limited. For labeled antibody binding to antigen on the immobilized particles, the detection limit is approximately 0.5 ng/mL. When antibodies on the immobilized particles exhibit multivalent binding of target molecules, the detection limit is approximately 0.01 ng/mL. For protein arrays, potential advantages of this format are improved coating of the capture reagent, an increased number of options for protein presentation, reduced mass transport effects, and higher density multiplexing. PMID- 12641233 TI - Microarrays assembled in microfluidic chips fabricated from poly(methyl methacrylate) for the detection of low-abundant DNA mutations. AB - Low-density arrays were assembled into microfluidic channels hot-embossed in poly(methyl methacrylate) (PMMA) to allow the detection of low-abundant mutations in gene fragments (K-ras) that carry point mutations with high diagnostic value for colorectal cancers. Following spotting, the chip was assembled with a cover plate and the array accessed using microfluidics in order to enhance the kinetics associated with hybridization. The array was configured with zip code sequences (24-mers) that were complementary to sequences present on the target. The hybridization targets were generated using an allele-specific ligase detection reaction (LDR), in which two primers (discriminating primer that carriers the complement base to the mutation being interrogated and a common primer) that flank the point mutation and were ligated joined together) only when the particular mutation was present in the genomic DNA. The discriminating primer contained on its 5'-end the zip code complement (directs the LDR product to the appropriate site of the array), and the common primer carried on its 3' end a fluorescent dye (near-IR dye IRD-800). The coupling chemistry (5'-amine containing oligonucleotide tethered to PMMA surface) was optimized to maximize the loading level of the zip code oligonucleotide, improve hybridization sensitivity (detection of low-abundant mutant DNAs in high copy numbers of normal sequences), and increase the stability of the linkage chemistry to permit re interrogation of the array. It was found that microfluidic addressing of the array reduced the hybridization time from 3 h for a conventional array to less than 1 min. In addition, the coupling chemistry allowed reuse of the array > 12 times before noticing significant loss of hybridization signal. The array was used to detect a point mutation in a K-ras oncogene at a level of 1 mutant DNA in 10,000 wild-type sequences. PMID- 12641235 TI - Imaging and analysis of immobilized particle arrays. AB - An automated imaging system was developed to quantify fluorescence signals from particles immobilized on hydrogel-coated slides. Arrays of submicrometer-diameter particles were printed with up to 600 particles/spot. The slides were read under 20x magnification without cover slips. Software was written to image individual spots and measure the median particle fluorescence in each spot. To locate array spots, an alignment program made use of two fiducial grids of fluorescent reference particles at either end of the slide. Focusing was adjusted locally using spots of reference particles located at the centers of focusing neighborhoods. The response was linear across a two-decade range, and the precision of readings was better than 5% down to approximately 1000 fluors/particle. Exposure times varied with signal intensity, reaching 1 s at the lowest levels of fluorescence. Data demonstrate feasibility for measuring fluorescence from immobilized particle arrays on an automated microscope with accuracy and precision similar to fluorescence measurements of microparticles with a flow cytometer. This work provides automation of imaging and analysis procedures necessary for development of immobilized particle arrays as an analytical platform that combines advantageous features of both planar and suspension arrays. PMID- 12641237 TI - Development of a methodology based on metal-catalyzed oxidation reactions and mass spectrometry to determine the metal binding sites in copper metalloproteins. AB - Efforts have been made to develop a method that uses metal-catalyzed oxidation (MCO) reactions and mass spectrometry (MS) to identify the binding site of copper in metalloproteins. This method uses MCO reactions to oxidize the amino acids in the metal-binding site and MS to identify the amino acids that have been oxidized. Several reaction conditions, including Cu(II)/ascorbate/O2, Cu(II)/O2/H2O2, and Cu(II)/ascorbate/O2/H2O2, have been tested at varying concentrations to find the optimum conditions for specific oxidation of only the amino acids bound to copper. For small peptides, such as angiotensin I (Agt I) and [Gln11]-amyloid-beta-protein fragment 1-16 (A beta(1-16)), the optimum conditions for specific modification involve the use of Cu(II)/ascorbate/O2. For a larger protein, azurin, the speed and specificity of the MCO reactions are enhanced by the presence of a relatively high concentration of ascorbate (100 mM) and a small concentration of H2O2 (1 mM). Optimized reaction conditions combined with MS/MS and MSn analysis on a quadrupole ion trap mass spectrometer allow the copper-binding sites to be specifically identified. For Agt I and A beta(1-16), the amino acids bound to copper can be identified without any false positives. For azurin, four of the five amino acids bound to copper are identified with one false positive. This false positive, however, corresponds to the oxidation of Met44, which is probably due to its susceptibility to oxidation and its proximity to the only residue not identified (i.e., Gly45). The results altogether suggest that MCO reactions and MS provide a very promising approach for identifying the amino acid residues bound to copper in metalloproteins. PMID- 12641236 TI - Statistical characterization of ion trap tandem mass spectra from doubly charged tryptic peptides. AB - Collision-induced dissociation (CID) is a common ion activation technique used to energize mass-selected peptide ions during tandem mass spectrometry. Characteristic fragment ions form from the cleavage of amide bonds within a peptide undergoing CID, allowing the inference of its amino acid sequence. The statistical characterization of these fragment ions is essential for improving peptide identification algorithms and for understanding the complex reactions taking place during CID. An examination of 1465 ion trap spectra from doubly charged tryptic peptides reveals several trends important to understanding this fragmentation process. While less abundant than y ions, b ions are present in sufficient numbers to aid sequencing algorithms. Fragment ions exhibit a characteristic series-specific relationship between their masses and intensities. Each residue influences fragmentation at adjacent amide bonds, with Pro quantifiably enhancing cleavage at its N-terminal amide bond and His increasing the formation of b ions at its C-terminal amide bond. Fragment ions corresponding to a formal loss of ammonia appear preferentially in peptides containing Gln and Asn. These trends are partially responsible for the complexity of peptide tandem mass spectra. PMID- 12641238 TI - Variable-gradient generator for micro- and nano-HPLC. AB - A new, simple device generates accurate nano- and microflow rate gradients from any conventional HPLC system. The core of the new device is represented by an electric-actuated, computer-controlled, multiposition HPLC valve. The valve hosts six reservoirs for as many different mobile-phase compositions of increasing strength. A low flow rate stream pushes the weakest solvent through the column as long as required and at the desired flow rate, until the chromatographic run is started. From this time on, the electric actuation allows one to select which reservoir will be on-line with the column and for how long, thus generating a specific solvent gradient, through a sequence of controlled segments of precise mobile-phase composition. This permits one not only to exactly reproduce the programmed slope but also to achieve different gradient shapes (i.e., linear, convex, concave) for different separation needs. The new device has proven to be reliable and reproducible even at the lowest flow rate tested (250 nL x min(-1)) and in different chromatographic conditions. PMID- 12641239 TI - On-chip coupling of isoelectric focusing and free solution electrophoresis for multidimensional separations. AB - We have developed an acrylic microfluidic device that sequentially couples liquid phase isoelectric focusing (IEF) and free solution capillary electrophoresis (CE). Rapid separation (<1 min) and preconcentration (73x) of species were achieved in the initial IEF dimension. Using full-field fluorescence imaging, we observed nondispersive mobilization velocities on the order of 20 microm/s during characterization of the IEF step. This transport behavior allowed controlled electrokinetic mobilization of focused sample bands to a channel junction, where voltage switching was used to repeatedly inject effluent from the IEF dimension into an ampholyte-based CE separation. This second dimension was capable of analyzing all fluid volumes of interest from the IEF dimension, as IEF was 'parked' during each CE analysis and refocused prior to additional CE analyses. Investigation of each dimension of the integrated system showed time-dependent species displacement and band-broadening behavior consistent with IEF and CE, respectively. The peak capacity of the 2D system was approximately 1300. A comprehensive 2D analysis of a fluid volume spanning 15% of the total IEF channel length was completed in less than 5 min. PMID- 12641240 TI - Homogeneous immunoassay for detection of TNT and its analogues on a microfabricated capillary electrophoresis chip. AB - A homogeneous immunoassay for TNT and its analogues is developed using a microfabricated capillary electrophoresis chip. The assay is based on the rapid electrophoretic separation of an equilibrated mixture of an anti-TNT antibody, fluorescein-labeled TNT, and unlabeled TNT or its analogue. The band intensities of the free fluorescein-labeled TNT and of the antibody-antigen complex reveal the relative equilibrated concentrations. Titration of the anti-TNT antibody with a fluorescein-labeled TNT derivative yields a binding constant of (3.9 +/- 1.3) x 10(9) M(-1). The dissociation rate constant of the complex is determined by kinetic capillary electrophoresis using a folded channel and a rotary scanner to interrogate the separation at multiple time points. The dissociation rate constant is found to be 0.035 +/- 0.005 s(-1), and the resulting binding rate constant is (1.4 +/- 0.7) x 10(7) M(-1) s(-1). Binding constants of TNT and five of its analogues are determined by competitive assays: TNT (4.3 +/- 2.6) x 10(8) M(-1); 1,3,5-trinitrobenzene (5.1 +/- 3.3) x 10(7) M(-1); picric acid (7.5 +/- 4.4) x 10(6) M(-1); 2,4-dinitrotoluene (7.9 +/- 4.0) x 10(6) M(-1); 1,3 dinitrobenzene (1.0 +/- 0.7) x 10(6) M(-1); and 2,4-dinitrophenol (5.1 +/- 3.0) x 10(4) M(-1). TNT and its analogues can be assayed with high sensitivity (LOD 1 ng/mL) and with a wide dynamic range (1-300 ng/mL) using this chip-based method. PMID- 12641241 TI - On-line immunoaffinity-liquid chromatography-mass spectrometry for identification of amyloid disease markers in biological fluids. AB - Disease-specific alterations in proteins and peptides such as the appearance of new isoforms, changed relative concentrations of known isoforms, or changed catabolism characterize the group of protein precipitation disorders collectively known as amyloidoses. The goal of this study was to develop an approach for isolating and characterizing the pool of isoforms of a polypeptide of interest from biological fluids for use in development of diagnostic markers and elucidation of pathogenesis. For this purpose, we employed an on-line immunoaffinity-liquid chromatography-mass spectrometry (IA-LC-MS) modular approach using antibodies binding populations of protein isoforms. In this system, crude biological samples, e.g., serum, may be injected and subjected to fast hands-off analysis. The setup consists of an optional preclear column for removal of unspecific binding components, an immunoaffinity column, a short cartridgelike reversed-phase column, and an electrospray time-of-flight mass spectrometer. We have tested the system for the automated analysis of three amyloid-related polypeptides, serum amyloid P component, amyloid beta-peptide, and beta2-microglobulin, and we show the feasibility of detection of altered isoforms or determination of relative abundance of isoforms of the proteins from serum or cerebrospinal fluid samples. For each new protein investigated, the only change needed in the system is a new antibody or antibody mixture and the selection of a reversed-phase cartridge of appropriate hydrophobicity. PMID- 12641242 TI - Pulsed excitation source multiplexed fluorometry for the simultaneous measurement of multiple analytes. Continuous measurement of atmospheric hydrogen peroxide and methyl hydroperoxide. AB - Presently, solid-state sources such as light-emitting diodes (LEDs) provide for intense, nearly monochromatic light. They are available over a broad range of emission wavelengths. Unlike incandescent and discharge lamps, LEDs can be turned on and off at high speeds. The resulting light pulses are highly reproducible. This allows the use of a single photomultiplier tube (PMT), often the most expensive component in a high-sensitivity measurement system, as a multiplexed detector with multiple, fiber-optic-coupled, fluorescence-detection cells excited by solid-state sources. A time resolution of 1 min is adequate in many continuous detection schemes. This enables multiple-channel single-detector multiplexed measurement without any loss of S/N. On the basis of this principle, we describe a new automated continuous instrument for the simultaneous measurement of atmospheric hydrogen peroxide and methyl hydroperoxide (MHP). A Nafion membrane diffusion scrubber (DS) is used with hematin-catalyzed oxidation of thiamine to thiochrome for the measurement of H2O2, and an expanded poly(tetrafluoroethylene) (ePTFE) DS is used with a H2O2 destruction catalyst and horseradish peroxidase catalyzed oxidation of thiamine to thiochrome for the measurement of MHP. The respective limits of detection are 25 pptv and 15 pptv. Design, performance details, and illustrative results from a field campaign (Philadelphia NEO3PS study, 2001) are presented. PMID- 12641243 TI - Development of a method for the determination of Fusarium fungi on corn using mid infrared spectroscopy with attenuated total reflection and chemometrics. AB - A novel method, which enables the determination of fungal infection with Fusarium graminearum on corn within minutes, is presented. The ground sample was sieved and the particle size fraction between >250 and 100 microm was used for mid infrared/attenuated total reflection (ATR) measurements. The sample was pressed onto the ATR crystal, and reproducible pressure was applied. After the spectra were recorded, they were subjected to principle component analysis (PCA) and classified using cluster analysis. Observed changes in the spectra reflected changes in protein, carbohydrate, and lipid contents. Ergosterol (for the total fungal biomass) and the toxin deoxynivalenol (DON; a secondary metabolite) of Fusarium fungi served as reference parameters, because of their relevance for the examination of corn based food and feed. The repeatability was highly improved by sieving prior to recording the spectra, resulting in a better clustering in PCA score/score plots. The developed method enabled the separation of samples with a toxin content of as low as 310 microg/kg from noncontaminated (blank) samples. Investigated concentration ranges were 880-3600 microg/kg for ergosterol and 310 2596 microg/kg for DON. The percentage of correctly classified samples was up to 100% for individual samples compared with a number of blank samples. PMID- 12641244 TI - Effect of caged fluorescent dye on the electroosmotic mobility in microchannels. AB - We report on measurements of electroosmotic mobility in polymer microchannels and silica capillaries with and without the addition of a caged fluorescein dye to the buffer. For PMMA microchannels, the mobility was found to increase from (2.6 +/- 0.1) x 10(-4) cm2 V(-1) s(-1) to (4.6 +/- 0.1) x 10(-4) cm2 V(-1) s(-1) upon addition of 1.2 mmol/L of caged dye. For PC microchannels, the mobility increased from (4.3 +/- 0.2) x 10(-4) cm2 V(-1) s(-1) to (5.4 +/- 0.1) x 10(-4) cm2 V(-1) s(-1) upon addition of caged dye. For PDMS microchannels, the mobility increased from (4.3 +/- 0.2) x 10(-4) cm2 V(-1) s(-1) to (6.4 +/- 0.5) x 10(-4) cm2 V(-1) s(-1) upon addition of caged dye. For fused-silica capillaries, the mobility ((5.5 +/- 0.2) x 10(-4) cm2 V(-1) s(-1)) was unaffected by the addition of the caged dye. PMID- 12641245 TI - Jet expansion module for liquid-sampled GC-discharge ionization detection of liquefied gases. AB - A rugged jet expansion module was designed, built, and utilized to sample liquefied gas for gas chromatography-discharge ionization detector (GC-DID) analysis. The module is inserted at an intermediate stage between a higher pressure sample cylinder and lower pressure gas injection manifold. The title jet module permits selection of liquid-phase or vapor-phase sampling by setting the valve-cylinder orientation up or down (relative to gravity). It consists of a four-way stainless steel cross, plus the following components connected to pipe thread ports: (1) mechanical pressure gauge, -15 to +30 psig; (2) threaded nut with 0.010-in. jet orifice (connect to sample cylinder valve); (3) bellows valve (connect to GC manifold); (4) bellows valve (connect to expansion cylinder). Making use of the jet module, measurement of air concentration in [15N]ammonia and sulfur hexafluoride (single-component) samples was found to be significantly lower in the liquid phase by GC-DID. This compact apparatus enables phase selective sampling, reduces primary manifold contamination, and improves operator safety. PMID- 12641246 TI - Examination of plant performance and filter ripening with particle counters at full-scale water treatment plants. AB - Filter performance has traditionally been assessed using turbidity as the main water quality parameter. However interest in process optimization and the ability to accurately monitor filtration and particle removal has led to particle counting becoming increasingly more popular. The objective of this research was to evaluate filtration performance and filter ripening at four full-scale water treatment plants. Most of the plants investigated in this study were capable of achieving 2 log total particle removal as well as an effluent turbidity of 0.1 NTU. In some cases 2 log removal was achieved in the lower particle size ranges of 2-5 and 5-10 microm. Log removals in this study compare particle counts in the filtered and raw water. For water treatment plants that do not filter to waste, the ripening period following a filter backwash represents a time where the plant may be most vulnerable to breakthrough of waterborne pathogens. For the plants studied, the ripening period which was defined as the time required to reach peak particle removal and a turbidity of 0.1 NTU, were very similar in duration. The ripening period generally lasted for one hour after the backwash event had been completed. Because the times to reach 2 log total particle removal and 0.1 NTU are similar, the practice of using 0.1 NTU turbidity as a measure of filter ripening may be a useful benchmark for utilities that do not have particle counters. PMID- 12641247 TI - Variation of pH, conductivity and potential values in chromium (VI) removal by wool. AB - Variation of pH, conductivity and potential values in chromium (VI) removal by wool was investigated in this study. In addition to chromium (VI) concentrations, pH, conductivity and potential values were also traced during the experimental studies. After applying linear regression to experimental data, it was seen that there was a relationship between pH, conductivity, potential values and chromium (VI) concentrations in the aqueous solution after treatment with wool. This relationship was evaluated in the light of the analyses of the linear regression. Kinetic data and adsorption isotherms for chromium (VI) adsorption by wool were also determined. PMID- 12641248 TI - Filamentous bulking control by sponge media in lab-scale SBR process. AB - Polyurethane foam sponge media were examined with respect to their ability to control filamentous bulking in a sequencing batch reactor process. In a high-rate nutrient removal study conducted with a 201 lab-scale reactor and synthetic wastewater of approximately 200 mgBOD 1(1), the presence of 20% v/v sponge media in the reactor improved the settling properties of biomass significantly, resulting in enhanced nutrient removal performance, i.e., < 10 mgT-N 1(-1) and < 1 mgT-P 1(-1) even at the HRT of 0.67 day. Without the support of sponge media, the suspended biomass was so bulky as to lead to its heavy washout from the reactor. A microscopic study under filamentous bulking revealed that the sponge media physically cut or broke biomass to shorter filaments and smaller flocs, mitigating the severe bulking condition. Indirect effects derived from the physical breakdown of biomass, such as more aerobic conditions in flocs, are expected to further create favorable conditions for suppressing filamentous bulking. PMID- 12641249 TI - Effect of organic waste amendments on degradation of PAHs in soil using thermophillic composting. AB - The feasibility of using various types of organic wastes including pig manure, sewage sludge, and soybean refuse for remediation of soil spiked with phenanthrene, anthracene and pyrene (PAHs) was evaluated through batch-scale composting reactors. The most active degradation of PAHs occurred between day 4 to 30 and maximum removal at the end of composting accounted for 90% of the initial concentrations of the three PAH compounds. Among the three PAHs, degradation of pyrene in the composting mass was relatively slow as indicated by a longer lag period than that of phenanthrene and anthracene. This corresponded well with the high molecular weight and log K(ow) values of pyrene. The organic amendments were effective in enhancing the degradation of PAHs, and pig manure amendment exhibited a slightly higher removal efficiency than sewage sludge and soybean refuse. A decrease in total organic matter in all treatments indicated that the decomposition process occurred. Toxicity test with cress seed germination was evaluated and no phytotoxicity was noted after 21 days of composting. This preliminary study positively supports that pig manure is an effective organic additive for bioremediation of PAH-contaminated soil using composting as a treatment technology. PMID- 12641250 TI - Comparison of suspended growth and hybrid systems for nitrogen removal in ammonium bisulfite pulp mill wastewater. AB - A series of bench-scale nitrification/denitrification tests were carried out with both suspended growth and hybrid bioreactors. The hybrid reactor was filled with plastic (polyethylene) media to evaluate the effects of biofilm. Two types of reactor configurations were tested; 4-compartment and 6-compartment modes. The experiments were initiated with a half-strength pulp and paper wastewater and its strength increased stepwise to the raw wastewater. Solid retention time was fixed at 10 days after a start-up period while hydraulic retention time was extensively varied from 3.5 to 0.5 days. The results from each type of reactor were compared in terms of nitrification/denitrification efficiency and stability. Experimental results demonstrate that the hybrid system showed greater stability in nitrification and higher denitrification efficiency than the suspended growth system. In the hybrid system, attached volatile solids formed 61- 72% of total volatile solids in the reactor and the amount of attached volatile solids insignificantly varied with the organic loading rate (0.37 - 2.76 kg COD M(-3) d( 1)) after initial biomass attachment. Under the conditions tested (0.1 - 2.8 kg COD m(-3) d(-1)), organic loading rate insignificantly influenced the nitrification. Better performance was obtained in denitrification when the anoxic zone was better isolated from the aerobic compartments (6-compartment mode). Overall, the hybrid system with fixed-film growth had better resistance to upset caused by transients such as changes in influent composition or hydraulic retention time. PMID- 12641251 TI - Biomass characterization of slow sand filtration schmutzdecke and its effects on filter performance. AB - A new approach of representing biomass by measuring the Total Kjeldahl Nitrogen (TKN) content of sand in the schmutzdecke of slow sand filtration (SSF) was established by correlating TKN, Volatile Solids (VS) and Total Plate Count, a better way for the representation of biomass against the usual method by measuring VS. Reasonably good correlations were found between TKN and Chemical Oxygen Demand (COD) and Biochemical Oxygen Demand (BOD) of the extract from the schmutzdecke sand. The effects of the concentration of TKN on the efficiency of the SSF used as a tertiary unit of a wastewater treatment plant in removing BOD, COD, Ammonia Nitrogen (NH(3)-N) and Total Microorganisms have been evaluated and better performances of the filter have been observed at higher TKN concentration. The removal of NH(3)-N was almost proportional to TKN concentration further proving the importance of measuring the NH(3)-N in the effluent to identify the degree of ripening of the filter. PMID- 12641252 TI - Integrated horizontal-flow anaerobic and radial-flow aerobic reactors for the removal of organic matter and nitrogen from domestic sewage. AB - This paper presents the conception and discusses the results obtained from the operation of an integrated biological anaerobic/aerobic/anaerobic system composed of horizontal-flow anaerobic and radial-flow aerobic reactors for domestic sewage treatment. The performance of a horizontal-flow anaerobic immobilized biomass reactor, with five stages,followed by a radial-flow aerobic immobilized biomass reactor was evaluated along 22 weeks. After the 14th week, the last stage of the HAIB reactor was used as a denitrifying unit. Polyurethane foam cubic matrices with 1-cm sides were used as support for biomass immobilization in all the units. The influent domestic sewage presented mean chemical oxygen demand of 365 +/- 71 mg. 1(-1) and the temperature was 23 +/- 3degrees C. The integrated system achieved COD removal efficiency of 90% while the maximum ammonium removal efficiency was 97% in the aerobic post-treatment unit. The nitrification process was found to be better represented by first-order reactions in series model. The apparent first-order kinetic coefficient for nitrate formation was about 50 times higher than that estimated for the nitrite formation. The denitrification process was well represented by a Monod-type kinetic model. The maximum specific denitrifying rate and the half-saturation coefficient were 2.9 x 10(-4) mg NO(3)( )-N mg(-1) VSS h(-1) and 19.4 mg NO(3)(-)-N 1(-1), respectively. PMID- 12641253 TI - Effects of partial stone removal on the co-composting of olive-oil processing solid residues with poultry manure and the quality of compost. AB - Full scale composting studies were performed with stoned (SEOOPSW) and non stoned exhausted olive-oil processing solid waste (NSEOOPSW) mixed with 20% poultry manure to reach a C/N ratio of around 30 and to improve the chemical and microbiological properties of the end products. The experiment was performed on two windrows (3 m wide, 2.5 m high and 10 tonne weight) to investigate the effects of stone removal and poultry manure addition on composting process efficiency and on the quality of composts. The partially stone removal operation reduced the total composting time by at least one month compared with the untreated waste. The addition of poultry manure improved the chemical quality of the end product, as shown by the doubling of the nitrogen concentration after four months composting together with the increased amount of P and K. Compost from stoned exhausted olive-oil processing solid waste had higher levels of cation exchange capacity (CEC) and humic acid content with a lower [FA/HA] ratio indicating a high polymerisation level. Agronomic field test showed that both composts are not phytotoxic. When applied at the rate of 40 tonne ha(-1) to a potato culture, they produced approximately the same yield (46 and 48.5 tonne ha( 1)) but they acted differently in terms of plant development. Compost produced from the NSEOOPSW acted more positively on the stem length (84.1 against 77.6 cm) and on leaf weight (354 against 238 g), whereas compost from the SEOOPSW more positively affected tuber sizes (76% large grade compared to 55%). PMID- 12641254 TI - Reductive dechlorination of TCE by zero valent bimetals. AB - Six zero valent bimetal combinations were tested for trichloroethene (TCE) dechlorination. Palladium, nickel, and copper were coated at 500 mg kg(-1) or less onto zero valent iron, producing Pd/Fe, Ni/Fe, and Cu/Fe bimetals, and zinc, producing Pd/Zn, Ni/Zn, and Cu/Zn bimetals. The order of reactivity of the six bimetals and two metals based on surface area normalized pseudo first order rate constants was observed to be: Pd/Fe > Pd/Zn > Ni/Fe > Cu/Fe > Ni/Zn > Cu/Zn > Fe > Zn. Thus, in general, 1) iron metal and bimetals exhibit faster reaction rates than the corresponding zinc reductants, 2) bimetals exhibit faster reaction rates than pure metals, and 3) metal coating enhance reactivity in the order of Pd > Ni > Cu. Chlorinated intermediate products were observed in small amounts for Fe only. In all cases, the chloride balance, calculated from TCE and chloride concentrations, ranged from 90 to 111%. The results are discussed in terms of the effects of bimetal combinations on catalytic dechlorination, prevention of nonreactive film and metal corrosion. PMID- 12641255 TI - Water flow patterns in subsurface flow constructed wetlands designed for on-site domestic wastewater treatment. AB - Bypass flow in subsurface flow constructed wetlands, which may be related to several wetland characteristics, reduces detention time and may result in inadequately treated wastewater. Subsurface flow constructed wetlands, 2.3 m wide by 4.8 m long and containing a gravel matrix, were used to investigate the impact of wetland depth, inlet pipe location, loading volume, and plants on water flow. Flow patterns were determined using blue dye or bromide as tracers. The blue dye adsorbed to the gravel and was not an effective tracer for following water movement. Water dispersed as it flowed through the wetland, and approximately two pore volumes of added water were required to displace approximately 99% of the bromide tracer added as a pulse. In 17 and 25 cm deep wetlands, water flow was uniform with depth, and inlet depth had little influence on water flow patterns. Water flow in a 40 cm deep wetland was not uniform with depth for either inlet pipe placement. The presence of plants caused preferential water flow around root masses, thereby, limiting their potential to interface with wastewater. Water mixing by dispersion from the surface to deeper depths may enhance aeration. Bromide was first detected in effluent after only 0.5 pore volumes of tap water had been added. This indicates that detention time for some wastewater would be less than expected, since plug flow is usually assumed in subsurface flow constructed wetlands. This occurrence should be considered for time-dependent treatments, such as fecal coliform and biological oxygen demand reduction. PMID- 12641256 TI - Removal of algae using froth flotation. AB - This paper describes the removal of algae and attached water using a froth flotation method as a function of the collector type, aeration rates, the pH of the algal suspension and temperature of operation. Dispersed air flotation was used in this study to remove Scenedesmus quadricaudo. The addition of surfactants such as cetyltrimethylammonium bromide and sodium dodecylsulfate increased the aeration rates and reduced the size of air bubbles. Only cetyltrimethylammonium bromide gave high algal removal (90%) whereas sodium dodecylsulfate gave poor algal removal (16%). However, by decreasing the pH values of the algal suspension, it was possible to increase the algal removal efficiency up to 80%. Low temperature operation had an important effect on reducing the rate of algal removal but when the temperature was 20 degrees C or higher there was little change with further temperature rises. The amount of water removed with the algal phase was reduced by using a two-stage flotation process. PMID- 12641257 TI - Evaluation of organic matter and nutrient composition of partially decomposed and composted spent pig litter. AB - Characterization of soil-applied organic material is necessary in order to clarify the nature of the organic matterand nutrients in it. In this study, the organic matter and nutrient contents of the spent pig litter (a mixture of partially decomposed pig manure and sawdust) was characterized before and after windrow composting to: (1) determine their changes during composting, and (2) assess the suitability of the composted spent litter as a soil amendment. Results demonstrated that the time required to reach maturity, and the composition of composted spent litter, depended on the chemical properties of the initial compost feedstock as well as the compost strategies used during composting. Total N, P, and K concentrations of the composted litter depended on chemical properties of the initial material. On the other hand, C:N ratio, humic and fulvic acid and cation-exchange capacity were influenced by differences in composition of the initial spent litter and composting strategy. If moisture content was maintained weekly at 60% with a four-day turning frequency, the litter reached maturity in 56 days. Maturation of spent litter was accompanied by a decline in total C, water-extractable metals, NH(4)(+)-N, increase in ash, (NO(3)(-)+NO(2)(-)-N, humic acid, humic acid:fulvic acid ratio, and cation exchange capacity, and elimination of phytotoxicity. The stability of nutrient and organic matter, acceptable pH and electrical conductivity values, and low levels of undesirable components such as heavy metals and phytotoxic compounds of the spent litter provided substantial evidence that agronomically suitable compost can be obtained after composting in windrows. PMID- 12641258 TI - Activated sludge acclimatisation kinetics to non-ionic surfactants. AB - The biodegradation of surfactants is a frequent and complex problem in domestic and industrial wastewater treatment processes. In addition to the resulting metabolites being sometimes refractory, the complete biodegradation of many of the most employed non-ionic surfactants requires long hydraulic retention times and the presence of specialised bacterial consortia. Preliminary acclimatisation tests highlighted the importance of the sludge acclimatisation state to a specific surfactant substrate for biotreatment efficiency. This paper reports on studies aimed at quantifying activated sludge acclimatisation and memory retention levels when subjected to changes in the type of surfactant included in the feed. Several transitions were tested, namely from an alkylphenol ethoxylate to a linear alkyl ethoxylate and the reverse, and between alkyl ethoxylates with different hydrophobic and hydrophilic molecular chain lengths. The kinetic results showed that sludge activation and memory loss were more dynamic for primary biodegradation It was found that the sludge was harder to adapt to alkylphenol ethoxylate than to alkyl ethoxylate. The former also apparently introduced an inhibitory effect, resulting in very slow degradation kinetics when imposed to alkyl ethoxylate acclimatised sludge. When replacing an alkyl ethoxylate with another surfactant of the same family, a longer ethoxylate chain reduced the degradation rates. This effect was further enhanced by simultaneously increasing the hydrophobic chain length of the substrate. The acclimatisation kinetic after the replacement of an alkyl ethoxylate by a longer counterpart was slower than the reverse case, and memory was also more easily lost. PMID- 12641259 TI - Pathogens and manure management systems: a review. AB - There has been an increasing concern about the effects of pathogens that are present in animal manure on humanand animal health. In recent years, outbreaks of food-borne diseases associated with the consumption of animal products havereceived much attention from the media in North America and Europe, leading to increased consumer concerns about the safety of their food supply. The health risks associated with animal operations depend on various factors. The most important ones appear to be related to the animal species being reared and the concentration of pathogenic microorganisms in animal manure. The ability of the pathogens to survive for long periods and through treatment to remain infective in the environment until ingested by human or animal host is an added concern. On the other hand, the role of livestock in most waterborne bacterial outbreaks has often been difficult to clarify since both humans and various wildlife species can shed the same microorganisms and thereby serve as sources of infection. This paper summarizes existing information on the main microbial pathogens present in livestock wastes, and discusses the impact of livestock wastes and agricultural drainage on microbiological quality of water, as well as available management and treatment technologies to minimize the prevalence of pathogens in animal wastes. Despite the fact that most disease outbreaks have been associated with food poisoning by cross-contamination during meat or milk processing and during finished product storage this review shows that a number of best management practices and technical solutions have been developed in the last few years that can be effective tools in minimizing the spread of pathogens from livestock operations in the environment. PMID- 12641260 TI - Reforming pharmaceutical policies in the European Union: a "penguin effect"? AB - Pharmaceutical policies form a substantial part of health care services, from the point of view of both equity and efficiency goals. Expenditure on pharmaceuticals has been growing steadily over the last few decades, and countries are finding the financing of drugs increasingly difficult. This article surveys the changes in pharmaceutical policies in the E.U. countries from the mid-1980s through the 1990s. It focuses primarily on policies dealing with cost control of publicly funded pharmaceuticals. In their analysis of these changes, the authors classify policies (or "packages of measures"), map out their incidence in each country, and assess their impact on the control of public pharmaceutical spending. They conclude that the E.U. countries are taking up apparently similar measures- dressing like penguins in a row--despite the limited effectiveness and limited evaluation of many of the measures adopted. The authors also analyze the role of national and international actors (most prominently, the European Union) in defining public pharmaceutical policies; look at how innovative policy ideas could be connected with the economic, political, and social interests that mold public action in this field; and propose new lines of investigation. PMID- 12641261 TI - Globalization of clinical research by the pharmaceutical industry. AB - Drug companies' quest for speedy results has led to a boom in trials based in developing countries, where ethical standards may be lax and the impoverished sick abundant. According to the U.S. Department of Health and Human Services Inspector General's office, the number of researchers based outside the United States seeking new drug approvals has increased 16-fold over the last decade. In this article, a 1996 Pfizer trial in Nigeria--the subject of a controversial class-action suit--illustrates the dangers. PMID- 12641262 TI - The paradoxical impact of health care restructuring in Canada on nursing as a profession. AB - This article draws on the concept of "countervailing powers" to explore some of the contradictory effects of Canadian health care restructuring on nursing. The main focus is on key institutional powers in the nursing field, the major individual and collective strategies nurses have adopted in response to restructuring, and the ways in which the interaction between global and national market forces and the aggregate responses of nurses has created a severe shortage of nurses. The global shortage has led to a global competition for nurses' labor. This, along with government budget surpluses, has increased nurses' bargaining power, forcing governments and hospital managers to reverse nursing spending cuts; to offer more secure professional jobs, as opposed to casual work; to engage in aggressive, bonuses-laden recruitment of nurses, both within Canada and abroad; and, more generally, to rethink some of their restructuring strategies. However, since the bargaining power of nurses is largely market dependent and, as such, highly variable, there does not seem to be much potential for a sustained increase in the institutionalized power of the nursing profession. PMID- 12641263 TI - Physician collective bargaining in a U.S. public hospital. AB - In a study to investigate the factors that would drive attending physicians employed in a public hospital to seek collective bargaining with their employer, the authors developed an instrument to determine which variables and which hypotheses were predictive of union proneness. The findings reveal that a desire for voice was the number one reason for physicians' wanting to join a union. Union-prone physicians had a lower salary on average, were more dissatisfied with their income, were more likely to feel the effects of work "speed up" (too many patients and too little time), were less likely to have administrative functions (thus a larger patient care role), had a strong sense of entitlement to collective bargaining, believed that unions improve participation in decisions affecting their jobs (reinforcing their desire for voice), and had a sense that a union would improve their treatment by supervisors (reinforcing their desire for due process and equity). PMID- 12641264 TI - Back to the market: yet more reform of the National Health Service. AB - Yet more reform of the National Health Service in England has been announced by the Department of Health. In opposition, the Labour Party criticized the creation of an "internal market" for health care by the Conservative government, but five years into the Blair administration, market incentives are to be reinvigorated and the private sector is to be embraced in ways not seen hitherto. New guidance signals the introduction of competitive contracting using cost-per-case currencies, more choice for patients in where they will receive hospital treatment, and the freeing of NHS care providers from the direct political control of ministers. It is intended that the monopolistic features of the NHS in England should give way to greater pluralism, in particular through contracts with privately owned health care organizations. However, there is little evidence to suggest that these policies will be effective, and a number of practical problems may obstruct implementation. PMID- 12641265 TI - Integrating primary care and public health: learning from the community-oriented primary care model. AB - Community-oriented primary care (COPC), a 50-year-old widely applied innovative approach to primary care development, seems to be the same combination of public health and general practice perspectives currently sought in the formation of primary care trusts in Britain's NHS. The article reviews the experience of implementing COPC methods, the outcomes, and the applicability to and implications for primary care policy, taking the current British reforms as an example. The COPC model has been developed mainly in underserved populations to integrate public health objectives and primary care through interdisciplinary approaches, with active involvement of the target population. COPC methods are time consuming, can create problems with professional boundaries, and are vulnerable to socioeconomic changes. They can also deliver complex packages of care for target populations, particularly in poor areas underserved by traditional medical services. British primary care reforms may be seen as an unplanned, uncontrolled, nationwide experiment in applying COPC methods. They differ from COPC as applied elsewhere because change has been introduced from above rather than below, into a well-developed primary care system rather than underserved communities. International experience suggests the need for attention to factors promoting and impeding success and to reliable outcome measures. If this experiment succeeds, COPC methodology may facilitate similar changes in other health care systems. PMID- 12641266 TI - The future of work environment reforms: does the concept of work environment apply within the new economy? AB - "Work environment," as a conceptual framework for reforming working life, may not be readily transferable from the tangible conditions of the industrial context in which it was conceived to the more flexible conditions of modern labor. Since the flexible conditions of work generally presuppose an increased responsibility and some sort of personal commitment on the part of the worker, isolating the environmental conditions from the personal abilities of the worker is becoming more difficult. As a consequence, only to a limited extent can the problems of modern labor be interpreted as work environment issues and subjected to work environment measures. With the propagation of flexible working conditions, work environment institutions and their reform ambitions will be passed by. And work environment reforms, rather than being a practical task of coordinating different protective measures at workplaces, will be reduced to an argument within the ideologically motivated rejection of an increasingly polarized labor market. PMID- 12641267 TI - Immunization campaigns and political agendas: retrospective from Ecuador and El Salvador. AB - Since the mid-1980s international donors have promoted vertical, campaign-based strategies to help improve immunization coverage in poor countries. National immunization days (NIDs) are currently in vogue and are prominent in the worldwide polio eradication efforts. In spite of their widespread use, campaigns that include NIDs have not been well evaluated for their effects on coverage, reduction in vaccine-preventable diseases, or effects on the health system. An assessment of the results of two such campaigns implemented in Ecuador and El Salvador shows limited impact on short-term coverage and questionable effects on long-term coverage and disease incidence. Although NIDs may have substantial short-term political benefits, the vertical approach can undermine provision of routine services by ministries of health and may be counterproductive in the long term. PMID- 12641268 TI - The health crisis in Argentina. AB - The health crisis in Argentina is part of the larger crisis that has resulted from a collapse in the country's economic and political systems. After a brief review of the country's history over the last century, from international success story to economic failure, the author explains the health crisis in particular and the social crisis in general in terms of failed neoliberal policies imposed on Argentina by the United States and International Monetary Fund through the mediation of the country's political class. PMID- 12641269 TI - If they get sick, they are in trouble: health care restructuring, user charges, and equity in Vietnam. AB - The transition from a centrally planned economy in the 1980s and the implementation of a series of neoliberal health policy reform measures in 1989 affected the delivery and financing of Vietnam's health care services. More specifically, legalization of private medical practice, liberalization of the pharmaceutical industry, and introduction of user charges at public health facilities have effectively transformed Vietnam's near universal, publicly funded and provided health services into a highly unregulated private-public mix system, with serious consequences for Vietnam's health system. Using Vietnam's most recent household survey data and published facility-based data, this article examines some of the problems faced by Vietnam's health sector, with particular reference to efficiency, access, and equity. The data reveal four important findings: self-treatment is the dominant mode of treatment for both the poor and nonpoor; there is little or no regulation to protect patients from financial abuse by private medical providers, pharmacies, and drug vendors; in the face of a dwindling share of the state health budget in public hospital revenues and low salaries, hospitals increasingly rely on user charges and insurance premiums to finance services, including generous staff bonuses; and health care costs, especially hospital costs, are substantial for many low- and middle-income households. PMID- 12641270 TI - Reflections of an Indian scholar. AB - In India, health programs have often been imposed on the deprived poor by a syndicate of foreign agencies and the local ruling class. During the first two decades after Independence, the political setting was somewhat conducive to scientific debates on the development of health services for India's people. The scenario changed radically during the next three and a half decades, when the country's ruling class became more oppressive and foreign agencies exerted increasing pressure to impose a prefabricated and scientifically suspect agenda of health services that were even more unequal and iniquitous to the interests of the deprived. With the help of themes from some of his major works, the author reflects on his experiences of the last half-century to make a case for using scientific critiques as an instrument for resisting foreign domination. PMID- 12641271 TI - A review of data on the U.S. health sector Fall 2002. AB - This report presents information on the state of U.S. health care in mid-2002. It provides data on the uninsured and underinsured and their difficulties in finding health care; the increasing costs of care; health, social, and economic inequalities; and the role of corporate money in health care. Information is also presented on mental health care, the hospital and pharmaceutical industries, Medicare HMOs, and the state of nursing. The author then provides updates on Congressional activity and the results of polls on matters of health, and some data on health care systems elsewhere in the world. PMID- 12641272 TI - Chromosome instability in cancer: how, when, and why? AB - The majority of malignant tumors exhibit complex genomic aberrations, and it has been suggested that an elevated rate of chromosome mutation is present in neoplastic cells. This chromosomal evolution is probably initiated at a preinvase stage and then proceeds continuously but at variable rates during tumor progression. Even though the initiating events may vary among neoplasms, the end point is often a similar set of cytogenetic and cytological events. These include an excessive shortening of telomeric repeats, disrupting the normal mitotic process and triggering chromosomal breakage-fusion-bridge events and gross genomic imbalances. Telomere dysfunction may also cause a failure of cytokinesis, resulting in tetraploidization and the accumulation of supernumerary centrosomes. In turn, this could cause multipolar cell division and gross aberrations in chromosome number. Several well-known cancer risk factors could contribute to these processes. For instance, breakage-fusion-bridge instability can be induced directly by clastogenic substances, whereas papilloma virus infection has been shown to dysregulate centrosome function. A prerequisite for the induction of chromosome instability through these mechanisms is partial inactivation of the DNA damage response and the mitotic checkpoint functions. Also, overexpression of telomerase appears to play a crucial role for moderating the rate of chromosomal evolution. PMID- 12641273 TI - Telomerase inhibitors: a new option for chemotherapy. AB - Telomerase is expressed in most types of tumors but not in most somatic cells. This observation has led to two hypotheses: (i) telomerase activity is necessary for the proliferation of cancer cells and (ii) telomerase inhibitors are a powerful strategy for cancer chemotherapy. The properties and function of human telomerase are reviewed briefly. The effects of telomerase inhibitors on telomere length and tumor cell proliferation are then described. Much evidence, both pro and con, has accumulated about the value of telomerase as a target for therapy. Resolving the controversy will require a thorough understanding of telomere biology and clinical trials designed around potent inhibitors. PMID- 12641274 TI - Heterogeneous pathogenicity of retroviruses: lessons from birds, primates, and rodents. PMID- 12641275 TI - Epstein-Barr virus and undifferentiated nasopharyngeal carcinoma: new immunobiological and molecular insights on a long-standing etiopathogenic association. AB - Undifferentiated nasopharyngeal carcinoma (UCNT) is characterized by its unique epidemiologic, immunobiologic, virologic, and clinicopathologic features. Aside from environmental risk factors and possible genetic susceptibility, infection by the Epstein-Barr virus (EBV) constitutes a well-documented link for the development of UCNT. However, despite the fact that UCNT is the human tumor associated most consistently with EBV, the role played by this virus in the pathogenesis of UCNT is still largely speculative and is a matter of ongoing debate. The purpose of this article is to review recent advances, particularly at the immunovirological and molecular levels, linking EBV infection to UCNT, and to discuss those aspects that may be of relevance for a better diagnosis and/or prognosis of this tumor, as well as for development of novel therapeutic strategies. PMID- 12641276 TI - Advances in immunotherapy for prostate cancer. AB - Prostate cancer is the most common malignancy in American men. Metastatic prostate cancer is incurable, with the currently best treatment, androgen ablation, being only palliative. Therefore, there is a need to develop new, more effective therapies against this disease. Multiple immunotherapeutic strategies are being explored for the treatment of prostate cancer, with the hope that such treatment will be more effective and have fewer side effects than current treatment options. Several immunotherapy strategies have been shown to be effective against prostate tumors in animal models, and many of these strategies are beginning to be tested in clinical trials for their efficacy against human prostate cancer. It is likely that effective treatment of prostate cancer will require the use of both immunotherapeutic and traditional approaches in multimodality treatments. In addition, for immunotherapy to be effective against prostate cancer, ways to overcome immune evasion and immunosuppression by the tumor cells will need to be developed. PMID- 12641277 TI - Role of CD4+ T lymphocytes in antitumor immunity. PMID- 12641278 TI - Comparison of preparative characteristics of micro open parallel plate separators and microbore columns for concentration of trace species by displacement chromatography. AB - The potential for preparative concentration of trace species by displacement chromatography in a micro open parallel plate separator (microOPPS) has been explored. A comparison of the performance of the microOPPS with micro open tubular columns (microOTC) has been presented. Using simulation models, the effects of operating and equilibrium parameters on throughput and yield have been determined. It is shown that the microOPPS can offer considerable advantages over the traditional microOTCs. Most significantly, throughputs can be enhanced by more than an order of magnitude in many cases. This is primarily due to the higher loading per cycle in the microOPPS, made possible by the ability to change the depth of the channel independent of the width. While yield is generally lower in the microOPPS, this effect can be overcome by the proper selection of operating conditions. PMID- 12641279 TI - Use of synthetic adsorbents in preparative normal-phase liquid chromatography. AB - Normal-phase liquid chromatographic separation using polymeric synthetic adsorbents was investigated. The retention behavior of dialkyl phthalates under a hexane-isopropanol eluent system revealed that both polystyrenic and polymethacrylic adsorbents can be used for normal-phase liquid chromatography, and that the polymethacrylic adsorbent has stronger retentivity than the polystyrenic adsorbent. Applicability of these synthetic adsorbents, especially the polymethacrylic adsorbents, was verified by the separation of polyunsaturated fatty acids, schizandrin in Schisandra chinensis fruit, tocopherols and tocotrienols in vegetable oils using a 10-microm adsorbent packed into an HPLC column. Furthermore, the separation was expanded to preparative scale by using polymethacrylic adsorbents with 17- and 31-microm fine grade particle sizes. The effect of loading volume on the retention or separation was also examined, and separation was maintained up to the loading of several grams per liter adsorbent. These results demonstrate that commercially available synthetic adsorbents possess considerable performance as normal-phase chromatographic media for more precise purification of bioactive compounds. including pharmaceuticals and nutraceuticals with scale-up possibilities. PMID- 12641280 TI - Linear two-step gradient counter-current chromatography analysis based on a recursive solution of an equilibrium stage model. AB - The implementation of gradients in continuously operated chromatographic counter current processes has recently attracted considerable interest as a method to improve the performance of this effective separation method. If liquid mobile phases are applied it is advantageous to set the solvent strength in the desorbent stream higher than that in the feed stream. As a consequence. the components to be separated are more retained in the adsorption zones and more easily eluted in the desorption zones. Due to the additional degrees of freedom the design and the optimization of such a two-step gradient counter-current process is difficult. In this paper a steady state equilibrium stage model is used to simulate the process under linear conditions. A simple solution of the underlying model equations is presented capable to describe efficiently the unit for large stage numbers typically encountered in chromatographic columns. Due to the rapidity of the algorithm developed a broad range of operating conditions can be evaluated systematically for different types of gradients. The impact of (a) the functional dependence of the adsorption equilibrium constants on the solvent composition, (b) the number of equilibrium stages and (c) the specification of purity requirements is illustrated and discussed based on results of parametric calculations. The results achieved emphasize the potential of two-step gradient counter-current chromatography. PMID- 12641281 TI - Modeling of adsorption in hydrophobic interaction chromatography systems using a preferential interaction quadratic isotherm. AB - A preferential interaction quadratic isotherm model for hydrophobic interaction chromatographic systems is presented in this paper. In this isotherm, the nonlinear effect of salt on the capacity factor is described using the preferential interaction model developed by Perkins et al. [J. Chromatogr. A, 766 (1997) 1]. This is then coupled with a quadratic nonlinear isotherm to describe nonlinear adsorption behavior at high solute concentrations. The resulting preferential interaction quadratic isotherm is examined for its ability to describe solute adsorption behavior under both linear and nonlinear conditions over a wide range of salt concentrations in HIC systems. The results indicate that this isotherm is well suited for predicting nonlinear adsorption behavior in HIC systems for both proteins and low-molecular mass HIC displacers. PMID- 12641282 TI - Studies on the long-term thermal stability of stationary phases in subcritical water chromatography. AB - The long-term thermal stability of five commercially available reversed-phase columns has been evaluated under subcritical water conditions (100 and 150 degrees C). The five columns included Zorbax RX-C8, Nucleosil C18 AB, Hypersil BDS C18, PRP-1 (poly(styrene-divinylbenzene)), and ZirChrom-PS (polystyrene) columns. Retention factors, plate numbers, and peak areas were monitored over a prolonged period of time. Comparing the three silica-based columns, the Zorbax RX C, column was the most stable followed by the Nucleosil C18, AB column. The Hypersil BDS C18 column was the least stable under subcritical water conditions. The ZirChrom-PS column was stable at 100 degrees C for at least 7600 column volumes. Of all five columns, the polymeric PRP-1 column was the most stable under subcritical water conditions. PMID- 12641283 TI - Design of a new, twelve-channel electrophoretic apparatus based on the Gradiflow technology. AB - The Gradiflow technology, originally designed to carry out binary, size-based and charge sign-based electrophoretic protein separations, has been extended to simultaneously obtain multiple protein fractions from a single electrophoretic separation. The separation unit of the new apparatus houses the anode and cathode compartments and up to twelve shallow separation compartments through which the background electrolyte solution that contains the separated protein fractions is recirculated. The separation compartments are formed from grids as thin as 1.2 mm and polyacrylamide membranes as thin as 0.15 mm, all with corresponding multiple inlet and outlet ports. The average pore size of the polyacrylamide membranes can be varied to permit passage of proteins in the 5000-800 000 molecular mass range. The electric field, orthogonal to the flow paths of the recirculated background electrolyte, selectively moves the sample components across the polyacrylamide separation membranes. Selective protein transport can be achieved by exploiting differences in either the relative size of the proteins or the charge sign of the proteins. The advantages of the new apparatus stem from the synergistic combination of the short electrophoretic transfer distances, high electric field strength, large effective surface areas of the separation membranes, and the great flexibility with which apparati containing one to twelve separation compartments can be created. PMID- 12641284 TI - Preparative-scale isoelectric trapping enantiomer separations. AB - The new Gradiflow BF200 IET unit, developed for isoelectric trapping protein separations has been modified and used to carry out preparative-scale enantiomer separations. Hydroxypropyl beta-cyclodextrin was used as the chiral resolving agent to induce an isoelectric point difference between the enantiomers. Three isoelectric membranes with isoelectric points below, in between and above the isoelectric points of the complexed enantiomers were used to trap the separated enantiomers in the anodic and cathodic separation compartments of the Gradiflow BF200 IET apparatus, respectively. The production rates were about 15 times higher than those previously obtained with another isoelectric trapping device and about 30% higher than those obtained in a continuous free-flow electrophoretic device operated in the isoelectric focusing mode. The remarkable separation speed observed in the modified Gradiflow BF200 IET unit is attributed to the favorable interplay of the short electrophoretic transfer distance, the high electric field strength and the large effective surface areas of the isoelectric membranes. PMID- 12641285 TI - Use of ultrasound to monitor the packing of large-scale columns, the monitoring of media compression and the passage of molecules, such as monoclonal antibodies, through the column bed during chromatography. AB - The novel use of ultrasound as a detector in pilot- and production-scale chromatography is described. The difficulties in packing production scale chromatography columns using an integral packing valve are reviewed. Results are presented from the packing of 400- and 600-mm diameter columns with various medias. From these results it is proposed that when packing large columns using a packing valve, for a given medium and column size, there is an ideal rate and pattern (or control "corridor") by which the bed builds in order to give optimum performance. Ultrasound was shown to be able to monitor the building of such a column bed as the medium was pumped into the column. It was found that the ultrasound detector was sensitive to bed compression, mobile phase composition and components such as acetone, albumin, casein and monoclonal antibodies while on the chromatography bed. This enabled the visualisation, by ultrasound, of these components as they were chromatographed through the column. PMID- 12641286 TI - Multiobjective optimization of simulated moving bed and Varicol processes using a genetic algorithm. AB - The size of the packing material, the total number of columns and the total feed concentration have significant impacts on the economics of a preparative chromatographic separation, through their effects on column pressure drop, column efficiency and thermodynamics. In this work, the role of these parameters on the performances of a simulated moving bed and a Varicol process is investigated on a chiral separation system from literature, using an equilibrium stage model. A multiple objective optimization technique based on a genetic algorithm is adopted, which allows to maximize simultaneously the purity of the extract and productivity of the unit. In this way, it is possible to optimize and compare the performances of both processes in a wide range of parameter values, so as to assess their relative potential under equally optimized conditions. The optimization results, i.e. the so-called Pareto sets, have been discussed in the frame of equilibrium theory and the roles of these three parameters have been clarified. PMID- 12641287 TI - Direct enzyme adsorption from an unclarified microbial feedstock using suspended bed chromatography. AB - Suspended bed chromatography (SBC) is a new hybrid technique concomitantly benefiting from batch adsorption, the process advantages of an enclosed system, and its compatibility with established commercial chromatographic contactors and adsorbents. SBC was evaluated in the anion-exchange capture and chromatographic fractionation of native glyceraldehyde-3-phosphate dehydrogenase (G3PDH) from the complex mixture of molecular and particulate moieties that constitute wet-milled bakers' yeast. Method scouting established operating conditions exploiting Whatman Express-Ion Exchanger Q at pH 7.5 and a disrupted biomass concentration equivalent to 3.5% (wet mass/v original intact cells). Partially purified G3PDH was recovered directly from the yeast disruptate in a scaled-down process developed at 1/756 process scale. This was used to establish operating parameters to facilitate process scale-up to exploit a 44 cm I.D. Millipore IsoPak column, 18 kg (swollen mass) of Express-Ion Q anion-exchange cellulose and 275 1 of 3.5% (wet w/v) bakers' yeast disruptate. The generic utility of SBC was demonstrated for direct product adsorption from feedstocks characterised by a modest content of bioparticulates (equivalent to < 4% (wet w/v) disrupted cells). Analyses illustrated an enrichment of G3PDH in respect of enzyme concentration and significant reduction in product turbidity and Pico-Green reactivity (correlated with double stranded (ds) DNA content). The application niche for this new approach to primary protein recovery is discussed with particular reference to the downstream processing of coarsely clarified whole broths, cell disruptates and biological extracts. PMID- 12641288 TI - Tobacco protein separation by aqueous two-phase extraction. AB - Tobacco has long been considered as a host to produce large quantity of high valued recombinant proteins. However, dealing with large quantities of biomass is a challenge for downstream processing. Aqueous two-phase extraction (ATPE) has been widely used in purifying proteins from various sources. It is a protein friendly process and can be scaled up easily. In this paper, ATPE was studied for its applicability to recombinant protein purification from tobacco with egg white lysozyme as the model protein. Separate experiments with poly(ethylene glycol) (PEG)-salt-tobacco extract and PEG-salt-lysozyme were carried out to determine the partition behavior of tobacco protein and lysozyme, respectively. Two-level fractional factorial designs were used to study the effects of factors such as, PEG molecular mass, PEG concentration, the concentration of phase forming salt, sodium chloride concentration and pH, on protein partitioning. The results showed that, among the studied systems, PEG-sodium sulfate system was most suitable for lysozyme purification. Detailed experiments were conducted by spiking lysozyme into the tobacco extract. The conditions with highest selectivity of lysozyme over native tobacco protein were determined using a response surface design. The purification factor was further improved by decreasing the phase ratio along the tie line corresponding to the phase compositions with the highest selectivity. Under selected conditions the lysozyme yield was predicted to be 87% with a purification factor of 4 and concentration factor of 14. From this study, ATPE was shown to be suitable for initial protein recovery and partial purification from transgenic tobacco. PMID- 12641289 TI - Interaction of immunoglobulin G with N,N,N',N' ethylenediaminetetramethylenephosphonic acid-modified zirconia. AB - Zirconia beads (25-38 microm in diameter) were modified with N,N,N'.N' ethylenediaminetetramethylenephosphonic acid to generate a pseudo-biospecific support, r_PEZ. To better understand the force of interaction between the IgG and the r_PEZ, the equilibrium dissociation constant (Kd) was determined by static binding isotherms, as a function of temperature and by frontal analysis at different linear velocities. Temperature had no significant impact on the maximum static binding capacity (Q(max)) and the equilibrium-binding constant (Kd), whereas pH and the salt concentration had a noticeable impact on both Q(max) and Kd values. Q(max) was found to be in the range of 55-65 mg IgG per ml of beads and unaffected by temperature. The maximum dynamic binding capacity (Qx) was found to be in the range of 20-12 mg IgG per ml of beads. The adsorption rate constant (ka) was determined by a split-peak approach to be between 982 and 32421 mol(-1) s(-1) depending on the linear velocity. Adsorption rate of IgG on r_PEZ was studied as a function of both feed concentration and linear velocity. The standard enthalpy and entropy values were estimated for the interaction of IgG with this novel support. The binding constants were also determined by modeling the batch protein-uptake data. PMID- 12641290 TI - Evaluation of protein-A chromatography media. AB - In process-scale antibody purification, protein-A affinity chromatography is commonly used as the initial purification step. In this paper, two different protein-A media were evaluated. These adsorbents have a porous glass backbone with different pore sizes: 700 A and 1000 A. Adsorption equilibrium data of human immunoglobulins on these media were measured via a batch technique and correlated using the Langmuir isotherm model. A larger static capacity was found for the smaller pore size material, which is probably a result of the larger specific surface area and associated higher ligand concentration. The protein uptake kinetics were also obtained via a stirred tank experiment using different initial protein concentrations. A surface layer model was used to represent the protein uptake by the media and to estimate values of a concentration-independent effective diffusivity within the particle. Experimental breakthrough curves were also obtained from packed beds operated under different conditions. Calculated breakthrough profiles were found to be in good agreement with the experimental results. Experimental breakthrough data were used to determine the dependence of the dynamic capacity of the media as a function of the fluid residence time. A larger dynamic capacity was also found for the smaller pore size media. The permeability of large scale packed beds was also reported and used in conjunction with the dynamic capacity to calculate the process production rate. PMID- 12641291 TI - Identification of protein A media performance attributes that can be monitored as surrogates for retrovirus clearance during extended re-use. AB - A potential safety concern in biotechnology purification schemes that employ re use of column media, often for large numbers of chromatography runs, is loss of the virus removal capacity of the chromatographic purification operation over time. To define chromatography performance attributes that best predict retrovirus clearance during extended re-use of protein A media, small-scale protein A columns were cycled 150 to 460 times using concentrates of murine hybridoma cell culture supernatants, standard low pH elution buffers and different cleaning solutions (6 M urea, 6 M guanidine, 100 mM NaOH or 500 mM NaOH). Load, flow-through and eluate samples were taken periodically and assayed for reverse transcriptase (RT, an enzyme component of retroviruses) activity, bovine IgG (a component of the culture media), genomic DNA, leached protein A, and mouse IgG. Under all cleaning conditions tested, the log,10 reduction value (LRV) of RT activity did not decrease and impurity co-elution did not increase during the 150 to 460 purification/cleaning cycles. In the two studies in which the columns were cleaned with NaOH, the chromatography performance attribute that best predicted the column media lifespan was column capacity, as measured by antibody (Ab) step yield and breakthrough. In both studies, Ab capture decayed in a biphasic manner starting at cycle 200 (100 mM NaOH) or cycle 50 (500 mM NaOH). For media cycled 300+ times using 6 M urea or 6 M guanidine cleaning buffers, column performance, including RT activity LRV, was more stable, although small upward trends in Ab breakthrough were evident. In summary, our studies identify Ab step yield and breakthrough as performance attributes that decay prior to retrovirus LRV when protein A media is multiply-cycled. Thus, we propose that virus removal validation studies should be performed on new media only and these attributes can be monitored during protein A unit operations in lieu of performing virus removal validation studies with cycled protein A media. PMID- 12641292 TI - Adsorptive membrane chromatography for purification of plasmid DNA. AB - Adsorptive membranes were investigated for the downstream processing of plasmid DNA by quantifying both separation efficiencies and adsorption uptake with the anion-exchange membranes. Separation efficiencies of the 10-ml Mustang-Q were measured using pulses of 6.1-kilo base pair plasmid DNA and lysozyme tracers, and comparing the responses for both conventional and reverse-flow operation. The plasmid exhibited nearly 200 plates/cm, almost as high efficiency as the protein despite the large difference in size. This behavior contrasts strongly with typical behavior for spherical porous particle packings, which predicted large decreases in efficiency with increases in tracer size. Batch adsorption isotherms for the 6.1-kilo base pair plasmid on small sheets of anion-exchange membranes at various ionic strengths showed high capacities for very large biomolecules. The maximum binding capacity for the membrane unit was calculated as 10 mg plasmid/ml, an order of magnitude greater than typical values reported for porous beads. PMID- 12641293 TI - Separation of vanadium isotopes by ion-exchange chromatography. AB - Cation-exchange displacement chromatography of VO2+ was carried out for studying vanadium isotope effects in carboxylate ligand-exchange systems. The heavier isotope 51V was enriched in the carboxylate complex solution. The isotope separation coefficients epsilon(= alpha-1) for 50V/51V were 2.2 x 10(-4) and 2.4 x 10(-4) for citrate and lactate systems at 298 K, respectively. These values are much larger than those obtained in a previous study on the malate system. The existence of binuclear complexes of VO2+ may create the conditions for larger isotope fractionation. From the viewpoint of the process development of isotope separation, the heights equivalent to a theoretical plate of these processes were analyzed and found to be very small in each system due to the homogeneous, small and highly porous resin used. Citrate may be better than the other tested systems for the vanadium isotope separation. PMID- 12641294 TI - Storytelling, marginality, and community in Australia: how immigrants position their difference in health care settings. AB - Stories of conflict with hospital services, medical mismanagement, and negative outcomes of procedures and treatment circulate within immigrant communities. While the interpretations of medical events are often based on misperceptions and misunderstandings, the stories have instructional value in that they explain an unfamiliar system to new immigrants and provide starting points for advocacy for improved services. Our analysis of gossip and storytelling among women from the Horn of Africa involves an examination of stories of "pork injections," rejection of "black babies," and clinical incompetence. The data are drawn from a study of reproductive health and reproductive rights that was conducted among refugee and immigrant women from Sahel African and Middle Eastern communities in Melbourne, Australia. PMID- 12641295 TI - Sexuality, AIDS, and the lures of modernity: reflexivity and morality among young people in rural Tanzania. AB - An understanding of young people's perceptions of AIDS and their sexuality is an essential precondition for the effective planning of AIDS campaigns in sub Saharan Africa. In examining how young Luo men and women in Tanzania describe their sexual behavior, I show that cultural conceptions of sexuality gender, and trust have an important impact on their actions. I also show that these conceptions have been rendered ambiguous by globalization, modernity, and by AIDS campaigns themselves. The values that are imparted to young people from family or peers often conflict with the preventive advice provided by both governmental and non-governmental organizations. However, by critically reflecting upon the ambiguities and inconsistencies in their lives, the young Luo have proven to be self-conscious actors and moral subjects who are actively involved in the process of social change. In the concluding section I suggest how elements of self critique and self-reflection, as well as the often differing perspectives and dilemmas experienced by young men and women, can be taken into account in order to make future educational campaigns more effective. PMID- 12641296 TI - Medicine for the heart: the embodiment of faith in Morocco. AB - "Medicine for the Heart" is a Moroccan way of describing the power of the Quran to heal the heart, an organ and symbol that links spiritual, emotional, and physical experience. The recitation of Quranic verses offers housewives living in southeastern Saharan Morocco a way to manage emotions associated with recent social changes--emotions that distress the heart and the body. This article is a preliminary examination of how, in everyday life, the phenomenology of faith illuminates an embodied structure of well-being, where spiritual experience merges with physical and emotional experience. Combining the perspectives of the anthropology of experience and social psychology, I highlight some of the spiritual aspects of well-being that emerge in the household production of health, particularly in relation to the core cultural values of unity and interiority. The concept of the spiritual body locates this experience inside the body (in this case radiating from the heart) rather than outside (i.e., in cognitive or symbolic space). PMID- 12641297 TI - The end of the art of obstetrics. Introduction. PMID- 12641299 TI - Obstetric perineal injury: risk factors and strategies for prevention. AB - Fecal incontinence due to anal sphincter injury is the most important consequence of perineal trauma at vaginal delivery and may be of muscular or neurological origin. The risk of sphincter injury is increased at first delivery and in association with instrumental assistance, prolonged second stage, occipito posterior position, and midline episiotomy, but is not predictable in individual cases. Injury can be prevented or minimized by enhancing uterine contractility during first labors, optimizing perineal repair technique, and by appropriate postnatal assessment of symptomatic women. PMID- 12641298 TI - The role of episiotomy in current obstetric practice. AB - This review traces the clinical use of episiotomy from the eighteenth century to the present and explains why the procedure has a restricted function in current practice. The types of episiotomy are described, and the controversies surrounding the sequelae associated with the procedure are explored. The modern indications for episiotomy including the procedure's place at operative vaginal delivery are discussed. Although the role of the episiotomy in modern obstetrics may be limited, the procedure is important in situations involving nonreassuring fetal status, shoulder dystocia, and perhaps operative vaginal delivery. The optimal type of episiotomy, if any, at forceps or vacuum delivery is yet to be determined. PMID- 12641300 TI - Cesarean section on demand. AB - The safety of cesarean section has improved dramatically over the past 50 years. During the past 20 years a greater awareness of and discussion about the symptomatic morbidity that can result for women following vaginal delivery has occurred and women's expectations for the outcome of pregnancy for them and their babies has increased. A culture of choice has been promoted in recent years, but contrary to the anticipated demand for less obstetric intervention by those promoting choice, there has been an increase in demand for delivery by cesarean section rather than the reverse. With the balance in favor of benefit for the baby from delivery by cesarean section, it is now difficult to sustain the argument favoring vaginal delivery rather than planned cesarean section, using maternal morbidity and mortality statistics. A critical evaluation of the costs indicates that there are probably few grounds for denying women their request for cesarean section for economic reasons. It seems likely, therefore, that in the near future those advising women on the options for delivery will need to ensure that the risks of vaginal delivery are explained as well as those for planned cesarean section. PMID- 12641301 TI - Breech vaginal delivery at or near term. AB - Three percent to 4% of term fetuses will be breech at delivery. Evidence from randomized controlled trials has found a policy of planned cesarean section to be significantly better for the singleton fetus in breech presentation at term compared to a policy of planned vaginal birth. However, some women may wish to avoid cesarean section and for others, cesarean section may not be possible. We undertook this review to identify factors associated with higher and lower risk of adverse fetal or neonatal outcome at term during vaginal breech delivery. We searched MEDLINE from 1966 to 2002 using the search terms vaginal breech delivery and breech presentation and retrieved all relevant articles. We also reviewed personal references and reference lists of articles retrieved. Women who are older or who have a fetus that is either in footling presentation, has a hyperextended head or is estimated to weigh <2500 g or >4000 g may be at higher risk of adverse fetal outcome. Prolonged labor or not having an experienced clinician at vaginal breech birth may also increase the risk. Women with a fetus in breech presentation at term should be offered the option of delivery by planned cesarean section and should be informed that this will reduce their risk of adverse fetal or neonatal outcome. Practitioners should develop and maintain skills at vaginal breech delivery for those women not wishing or not able to be delivered by cesarean section. PMID- 12641302 TI - Controversies in the use of the vacuum extractor. AB - Operative pelvic delivery is an important component of obstetrical care. Vacuum extraction assumes a prominent role, and when appropriately performed, has been proven safe and effective. However, controversies continue to exist. Historical background and review of the latest literature are presented to delineate these issues and promote consensus and direct research to continue to provide the safest means of delivery for the mother and baby. PMID- 12641303 TI - Intrapartum management of multifetal pregnancies. AB - Over the past several decades advances in assisted-reproductive technologies have resulted in a dramatic increase in the number of multifetal gestations. Concomitant with this increase there has been a gradual rise in the overall preterm birth rate, as well as other pregnancy complications related to these pregnancies. Twin, triplet, and other high-order multifetal gestation pregnancies pose a number of important issues related to antepartum and intrapartum management. Antepartum issues include ultrasound determination of zygosity, management and prevention of preterm labor, maternal/fetal surveillance for complications, and specific interventions focused on prevention of adverse maternal and/or fetal outcomes. Intrapartum issues include those related to timing of delivery, labor management, anesthesia options, and determination of an optimal delivery modality. Clearly, these issues related to the management of multifetal pregnancies are of paramount importance to optimize pregnancy outcome. As many of the issues related to antepartum care for women with multifetal gestations have been reviewed elsewhere, we have restricted the focus of this article to intrapartum management. Thus, this article reviews salient issues related to the intrapartum management of multifetal gestations, including twins, triplets, and other high-order pregnancies. PMID- 12641304 TI - Current controversies in cervical cerclage. AB - Cervical incompetence is defined as the inability to support a full-term pregnancy because of a functional or structural defect of the cervix. It is characterized clinically by acute, painless dilatation of the cervix usually in the mid-trimester culminating in prolapse and/or premature rupture of the membranes with resultant preterm and often previable delivery. Cervical cerclage has become the mainstay for the management of cervical incompetence, but remains one the more controversial surgical interventions in obstetrics. This article reviews the current state of the literature as regards the indications, contraindications, and techniques of cervical cerclage. This article also focuses in detail on 4 areas of controversy, namely transabdominal cerclage, cervical cerclage for a short cervix, the management of cerclage after preterm premature rupture of the membranes, and the utility of a second (salvage) cerclage. PMID- 12641305 TI - Management of obstetric hemorrhage. AB - A reluctance to proceed with hysterectomy for obstetric hemorrhage may be a more likely cause of preventable death in obstetrics than a lack of surgical or medical skills. Every obstetric unit should have protocols available to deal with hemorrhage and, in addition, have specific guidelines for patients who object to blood transfusions for various reasons. Risk factors for hemorrhage should be identified antenatally, using all possible imaging modalities available, and utilizing multidisciplinary resources whenever possible. Novel strategies for prenatal diagnosis of abnormal placentation include advanced sonography and magnetic resonance imaging. Placement and utilization of arterial catheters for uterine artery embolization is becoming more widespread and new surgical technology such as the argon beam coagulator seems promising. When intra or postpartum hemorrhage is encountered, a familiar protocol for dealing with blood loss should be triggered. Timely hysterectomy should be performed for signs of refractory bleeding. Application of medical and surgical principles combined with recent technologic advances will help the obstetrician avoid disastrous outcomes for both mother and fetus. PMID- 12641306 TI - VBAC--is it worth the risk? AB - Enthusiasm for vaginal birth after cesarean section has waned. As a result, the cesarean birth rate is again on the rise. As a medical community and society we must decide whether the most appropriate question is "What is safest for my baby?" or "Is the risk associated with vaginal birth after cesarean acceptable?" There are risks associated with vaginal birth after cesarean, but in a hospital setting with appropriate resources these risks are low and would still seem to be acceptable. PMID- 12641307 TI - Rotational forceps: should these procedures be abandoned? AB - Forceps delivery remains an important part of the obstetric armamentarium. When applied by practitioners skilled in their use, forceps delivery can quickly and safely deliver a fetus at risk. Unfortunately, forceps can also be an instrument of harm for the women or her infant. This is particularly true of rotational forceps. The goal of this monograph is to review in detail the indications, contraindications, technique, as well as risks and complications of forceps delivery, with particular attention to rotational forceps. We conclude by asking the question: Should rotational forceps be abandoned altogether? PMID- 12641308 TI - The role of self-enhancing evaluations in a successful life transition. AB - The objective of the study was to investigate how self-enhancing evaluations, obtained via positive social comparisons and reflected appraisals, were related to mental health in a later life transition. The sample consisted of 266 women who were interviewed once before and 3 times after the experience of community relocation. Results extended prior findings suggesting the dynamic impact of self enhancing evaluative processes on psychological well-being and depressive symptoms. Additional analyses showed evidence for reverse causality, that is, that mental health also influenced self-evaluative processes. This study underscores the significance of holding a positive view of self vis-a-vis others in negotiating life challenges as well as the reciprocal influence of well-being on social self-evaluative processes. PMID- 12641309 TI - Looking back and looking ahead: adult age differences in consistency of diachronous ratings of subjective well-being. AB - The goal of this study is to investigate the consistency of diachronous ratings of subjective well-being (SWB). A heterogeneous sample (25-74-year-olds; N = 3,596) provided ratings of their present SWB, reconstructed their SWB of 10 years ago, and anticipated their SWB 10 years from now. Developmental tasks and self evaluative principles were used to predict age differences in diachronous consistency. As predicted, in young adulthood, past SWB was rated lower and future SWB higher than present SWB. In contrast, in later adulthood, the past was rated higher and the future lower than present SWB. Analyses of rank-order consistency demonstrated that in later adulthood both future and past SWB were more strongly related to present SWB than in young adulthood. Results show how models of self-evaluation play out at different points in the life span. PMID- 12641310 TI - Personality is associated with perceived health and functional status in older primary care patients. AB - Using data collected on 265 primary care medical patients 60 years of age and older, the authors examined the personality bases of subjective health (perceived health, functional status) after controlling for observer-rated depression and medical burden. Four hypotheses were tested: High Neuroticism is associated with poorer perceived health, low Extraversion is associated with poorer perceived health, low Openness to Experience is associated with worse functional status, and age moderates the relationships between personality and subjective health. Findings supported the notion that personality is associated with subjective health; moreover, this effect appeared to grow more pronounced with increasing age. This study underscores the conceptual and heuristic value of examining moderators of the links between personality variables and health. PMID- 12641311 TI - Longitudinal hierarchical linear models of the memory functioning questionnaire. AB - Three hypotheses about the nature of self-rated memory as measured by the Memory Functioning Questionnaire (MFQ; M. J. Gilewski, E. M. Zelinski, & K. W. Schaie, 1990) were tested: that ratings reflect memory performance, that personality traits underlie ratings, and that ratings reflect implicit theories of memory change. Baseline scores and 19 year change slopes for the 4 MFQ factor ratings of a sample of 97 participants aged 30-81 were investigated. There were significant mean declines for all MFQ ratings except Frequency of Forgetting and significant individual differences in slopes for Frequency, Retrospective Functioning, and Mnemonics. Personality predicted baseline Frequency and Seriousness ratings and list and text recall slopes predicted Mnemonics slopes. Different mechanisms may underlie baseline ratings and changes in ratings for different factors. PMID- 12641313 TI - Perceived self-efficacy and everyday problem solving among young and older adults. AB - This research tested the hypothesis that age differences in both self-efficacy perceptions and problem-solving performance would vary as a function of the ecological relevance of problems to young and older adults. The authors developed novel everyday problem-solving stimuli that were ecologically representative of problems commonly confronted by young adults (young-adult problems), older adults (older adult problems), or both (common problems). Performance on an abstract problem solving task lacking in ecological representativeness (the Tower of Hanoi problem) also was examined. Although young persons had higher self-efficacy beliefs and performance levels on the Tower of Hanoi task problem and the young adult problems, this pattern reversed in the domain of older adult problems, where the self-efficacy beliefs and performance of older persons exceeded those of the young. PMID- 12641312 TI - Age-related changes in selective attention and perceptual load during visual search. AB - Three visual search experiments were conducted to test the hypothesis that age differences in selective attention vary as a function of perceptual load (E. A. Maylor & N. Lavie, 1998). Under resource-limited conditions (Experiments 1 and 2), the distraction from irrelevant display items generally decreased as display size (perceptual load) increased. This perceptual load effect was similar for younger and older adults, contrary to the findings of Maylor and Lavie. Distraction at low perceptual loads appeared to reflect both general and specific inhibitory mechanisms. Under more data-limited conditions (Experiment 3), an age related decline in selective attention was evident, but the age difference was not attributable to capacity limitations as predicted by the perceptual load theory. PMID- 12641314 TI - Depression in later life: cross-sequential patterns and possible determinants. AB - Research on the relationship between age and depression has not yielded a consistent picture. Approaches emphasizing resilience compete with assumptions of an increased incidence of depressive problems in later life. This study investigated these issues within a research format combining cross-sectional and 8-year longitudinal assessments of depressive tendencies (690 participants, aged 54-77 years). The developmental patterns suggest 2 phases: Relative stability extending to about 70 years and a subsequent increase of depressive tendencies. Antecedent and mediating factors were explored within a broader context of variables (health status, social and economic resources, coping competences, and future perspectives). Differences in time perspective and a disposition of accommodative flexibility predicted subsequent changes in depression and mediated age effects in depressive tendencies. PMID- 12641315 TI - What needs to be explained to account for age-related effects on multiple cognitive variables? AB - Structural equation methodology was used to investigate age-related influences across a number of cognitive variables in 204 adults ranging from 18 to 91 years of age with a hierarchical structure that contained 4 1st-order factors and 1 2nd order common factor. Direct age relations were found to the common factor as well as to 1st-order speed and memory factors. Replicability of the findings was explored by investigating the same structure of age relations, using 2 different data sets, and a similar patten was found in each. These results suggest that at least 3 statistically distinct types of age-related influences are operating on a wide variety of cognitive variables and presumably require separate explanatory mechanisms. PMID- 12641316 TI - Memory performance and beliefs as a function of goal setting and aging. AB - This research examined the impact of goals on memory and memory beliefs. Older and younger adults completed memory beliefs questionnaires and list recall at baseline. After additional recall trials, the questionnaires were repeated. In Experiment 1, participants were assigned to low challenge or high challenge goals. In Experiment 2, moderate challenge goals were compared to control. In both studies, participants were given a specific goal based on their own performance and received positive feedback for memory gains. Both older and younger adults responded to the goals, showing improved performance across trials, with little change in the control condition. Memory beliefs changed in the moderate and low challenge goal conditions, showing more striking changes for the older groups. These results confirmed that self-regulatory processes related to goal setting can have considerable impact on memory across the adult life span. PMID- 12641317 TI - Breaking the waves: age differences in electrical brain activity when reading text with distractors. AB - The authors tested whether older adults have greater difficulty than younger adults in ignoring task-irrelevant information during reading as a result of age related decline in inhibitory processes. Participants were shown target sentences containing distractor words. They were instructed to read aloud each sentence and ignore distractors. The N400 event-related potential (ERP) was used to measure the extent of semantic processing of target and distracting information. It showed that younger adults semantically processed both target and distracting material, whereas online processing of target sentences in older adults was disrupted by the distractors. In older adults, memory for target information related to their susceptibility to distraction and inhibition efficiency. Implications for age-differences in inhibitory control, working memory, and resource capacity are discussed. PMID- 12641318 TI - Cerebral white matter abnormalities and lifetime cognitive change: a 67-year follow-up of the Scottish Mental Survey of 1932. AB - Cerebral white matter abnormalities relate to cognitive functioning in elders. We examine whether this association is (a) independent of mental ability in youth and (b) related to general and/or specific mental abilities. We retested 83 participants of the Scottish Mental Survey of 1932 on a battery of mental tests. Their brains were scanned by magnetic resonance imaging. Three independent ratings (Fazekas) were made of periventricular, and subcortical and deep white matter abnormalities. Structural equation models showed that, irrespective of brain location, white matter abnormalities contributed about 14% of cognitive function variance in old age. Some of this effect might be due to hypertension. This contribution is independent of mental function in early life and is associated with general cognitive ability. PMID- 12641319 TI - Selective adult age differences in an age-invariant multifactor model of declarative memory. AB - Confirmatory factor analysis was used to test competing models of declarative memory. Data from middle-aged participants provided support for a model comprised of 2 2nd-order (episodic and semantic memory) and 4 1st-order (recall, recognition, fluency, and knowledge) factors. Extending this model across young old and old-old participants established support for age invariance. Tests of group differences showed an age deficit in episodic memory that was more pronounced for recall than for recognition. For semantic memory, there was an increase in knowledge from middle to young-old age and thereafter a decrease. Overall, the results support the view that episodic memory is more age sensitive than semantic memory, but they also indicate that aging has differential effects within these 2 forms of memory. PMID- 12641320 TI - Aging and functional spatial relations in comprehension and memory. AB - Previous research has suggested that, relative to younger adults, older adults devote a greater proportion of their discourse processing to the situation model level. The current experiment assessed whether this is due, in part, to a preserved ability to focus on functionally appropriate information. The focus here was on spatial relations. Both reading time and recognition data showed superior performance for functional over nonfunctional information, and this functional effect was similar in younger and older adults. This is consistent with the idea that older adults' ability to process information at the situation model level is relatively well preserved. PMID- 12641321 TI - Antimycobacterial 3-aryl-2H-1,3-benzoxazine-2,4(3H)-diones. AB - A series of 153 derivatives of 3-phenyl-2H-benzoxazine-2,4(3H)-dione substituted in position 6 or 7 on benzoxazine and on the phenyl ring was synthesized. The compounds were evaluated in vitro for antimycobacterial activity against Mycobacterium tuberculosis, Mycobacterium kansasii and Mycobacterium avium. The activity of the compounds increases with increasing hydrophobicity and electron withdrawing properties of the substituents on the phenyl ring, whereas the effect of the substituents on the benzoxazine ring seems to be more complex. PMID- 12641322 TI - [New findings on the synthesis of the centrally acting muscle relaxant chlormezanone and its resolution of a gram scale using a Chiralcel OD column]. AB - 2-(4-Chlorophenyl)-4-metathiazanone (2) is the intermediate product for the two step-synthesis of chlormezanone (1), a centrally acting muscle relaxant. The second step includes the oxidation of its sulfur atom. It has been found that the foregoing reaction of 4-chlorobenzaldehyde with methylamine forming the hemiaminale and the subsequent addition of beta-mercaptopropionic acid leads to a remarkable better yield (67% of th.) than the route via the hemimercaptale (42% of th.). 2 could be oxidized with sodium perborate superior to potassium permanganate. The racemic chlormezanone (1) is resolved quickly on a gram scale by preparative column chromatography on a Chiralcel OD column (tris(3,5-dimethyl phenyl-carbamoyl)cellulose on silicagel). The resolution needed only 40 min, if flow rate, composition of the mobile phase and temperature as the most important factors are determined prior with an analytical column. Both dissociation constants could be determined for the first time with the aid of a log pKa Titrator of the Sirius Co., which needs for the registration of the curves only 15-17 min in the pH range of 2-12. This speed outplayed the disturbing cleavage of the S-C bond of chlormezanone at strong acidic and alkaline pH values. PMID- 12641323 TI - Prodrugs and mutual prodrugs: synthesis of some new pyrazolone and oxadiazole analogues of a few non-steroidal anti-inflammatory drugs. AB - Naproxen, probenecid, diclofenac, ibuprofen and indomethacin were converted to hydrazide derivatives via their methyl ester by reacting with hydrazine hydrate, which were further condensed with beta-keto esters to get the pyrazolone derivatives. The hydrazide derivatives of probenecid and diclofenac were also reacted with biphenyl acetic acid while naproxen hydrazide was reacted with p chloro benzoic acid besides biphenyl acetic acid to synthesize their oxadiazole analogues. Some selected members of the compounds prepared were screened for their anti-inflammatory and analgesic activity. PMID- 12641324 TI - [Reactions of isopropyl-alkylamine with 2,3-dichloro-1,4-naphthoquinone and 2,3 dichloro-1,4-naphthoquinone/acetaldehyde]. AB - Isopropyl-alkylamines 2 react with 2,3-dichloro-1,4-naphthoquinone (1) to give red 2-chloro-3-isopropyl-alkylamino-1,4-naphthoquinones 3 and with 2,3-dichloro 1,4-naphthoquinone/acetaldehyde to give blue 2-chloro-3-isopropylalkylamino-vinyl 1,4-naphthoquinones 7. It is evident that the formation of 7 is preferred sterically to the formation of 3. The reaction between 2, 1 and acetaldehyde give also red aminoquinones 3 and blue green 2-chloro-3-(4-isopropylalkylamino-buta 1,3-dienyl)-1,4-naphthoquinone like 11. PMID- 12641325 TI - Coupled column chromatography for separation and determination of enantiomers of phenylcarbamic acid derivatives in serum. AB - Columns packed with vancomycin coupled to an achiral C18 column and beta cyclodextrin were used for the separation and the determination of enantiomers of alkoxysubstituted esters of phenylcarbamic acid in blood serum. The method involves off-line SPE, the separation of the racemate on a reversed-phase stationary phase, and the separation of the enantiomers on a chiral stationary phase. The limit of detection was 1.0 microg/ml for the vancomycin column and 10.0 microg/ml for the beta-cyclodextrin column in standard solution. In vitro degradation studies of enantiomers demonstrated a difference in the concentation of the enantiomers after the treatment. It was found that the rate constants of R(-)- and S(+)-forms of enantiomers are not significantly different. PMID- 12641326 TI - Capillary isotachophoresis of cystine in urine with on-line isotachophoresis sample pretreatment. AB - Capillary isotachophoresis (ITP) with on-line isotachophoresis sample pretreatment and conductivity detection has been proposed as a method for the separation, detection and determination of cystine present in urine. Sample pretreatment was achieved using a 90 mm FEP capillary tube with a 800 microm internal diameter. ITP separations and conductivity detection were carried out in a 160 mm capillary tube with a 300 microm internal diameter, filled with a hydrochloric acid leading electrolyte and glycine terminating electrolyte, at 50 microA and 25 degrees C. The results show that the proposed method is suitable for resolution of cystine from matrix interferents present in the urine samples and is able to detect it at 10(-4) mol/l concentrations by a universal detection technique after removal of a significant part of the anionic constituents of the sample migrating in the on-line coupled capillary isotachophoresis stack. This simple ITP procedure for monitoring cystine in urine samples requires no sample preparation. This makes it an alternative to the currently used procedures. PMID- 12641328 TI - Developing a discriminating dissolution test for three mebendazole polymorphs based on solubility differences. AB - Mebendazole, a broad spectrum anthelmintic drug, is practically insoluble in water and exists in three polymorphic forms, A, B, and C, of which C is pharmaceutically favoured. Since the dissolution of drugs from solid oral dosage forms can depend on the crystal form of the drug an attempt should be made while developing dissolution tests to set test parameters that are sensitive to changes in the crystal form. USP 24 describes 0.1 M hydrochloric acid containing 1.0% sodium lauryl sulphate (SLS) as the dissolution medium for mebendazole tablets. Results showed that the high concentration of sodium lauryl sulphate in the USP dissolution medium does not allow the use of this test to distinguish between the solubility differences of the three mebendazole polymorphs. By decreasing the amount of sodium lauryl sulphate in the dissolution medium clear differences in the dissolution rates of the three forms were observed. The most discriminating medium was 0.1 M HCl, containing no sodium lauryl sulphate. PMID- 12641327 TI - Kinetic-spectrophotometric determination of propylthiouracil based on its inhibitory effect on the reduction of neutral red by hypophosphite. AB - A new kinetic-spectrophotometric method for determination of propylthiouracil (PTU) is described. The proposed method is simple, rapid, inexpensive and sensitive for the determination of PTU in pure and tablet forms. This method is based on the inhibitory effect of propylthiouracil on the palladium(II)-catalyzed reaction between neutral red and hypophosphite ions. The effect of various parameters such as: dye, hypophosphite, and Pd(II) concentrations, pH, ionic strength, and temperature were optimized. Two distinct linear calibration graphs were observed in the ranges 0.006-0.033 (n = 6, r = 0.9991) and 0.033-0.300 ppm (n = 10, r = 0.9980). The variable time method was used. The limit of detection was 0.004 ppm. The proposed method was applied for the determination of propylthiouracil in pure and tablet forms. PMID- 12641330 TI - Application of spectrofluorometry for evaluation of dry powder inhalers in vitro. AB - Andersen cascade impactor (ACI) is commonly used for the testing of pharmaceutical aerosols, which has to be coupled with an instrument for quantitative analysis of drug depositing on each stage of the ACI. This procedure consumes much time in operation. Therefore, this study was aimed at speeding up the process of drug analysis in aerosol formulations after obtaining samples from the ACI. From the results obtained, it was proved that the validated spectrofluorometric method was accurate and sensitive. It was capable of giving similar results to those we obtained from HPLC-UV analysis. There was no interference from the amount of lactose carrier incorporated in the formulation in the step of salbutamol analysis indicating specificity of the method. As a result, samples were analyzed without further separation. The detection limit was 0.1 microg/ml. Hence, spectrofluorometry can be used as a substitute method to HPLC-UV in determining the small quantity of salbutamol after aerosolization from dry powder aerosols. The present study suggests that spectrofluometry can be a rapid and efficient method in the pharmaceutical analysis of aerosols. PMID- 12641329 TI - Alkaline hydrolysis of cladribine. AB - The kinetics of hydrolysis of 2-chloro-2'-deoxyadenosine (cladribine) was studied at various sodium hydroxide concentrations and temperatures. HPLC analysis of reaction mixtures showed that the main products were 2'-deoxyisoguanosine and 2' deoxyguanosine. The first one was the result of the hydroxyl anion attack, whereas the presence of the other nucleoside has evidenced the existence of hitherto undescribed rearrangement reaction in purine derivatives. PMID- 12641331 TI - Development of indomethacin Carbopol ETD 2001 gels and the influence of storage time and temperature on their stability. AB - We investigated the development of a topical indomethacin gel formulation of suitable consistency using Carbopol ETD 2001 as the gelling agent. Topical gel formulations containing 1% w/w indomethacin (IND), 1% w/w Carbopol ETD 2001 (C2001), 1% of trethanolamine (TEA), 30% hexylene glycol (HG) and 10% polyethylene glycol (PEG 300) were prepared with excipients Tween 80, PVP 25, both Tween 80 and PVP 25 or neither agent. These four gel formulations were tested after a period of 1 and 4 weeks at storage temperature of 6 degrees C, 20 +/- 2 degrees C and 45 degrees C. Physical evaluation of the stability of these gels was carried out by microscopic and rheological tests, measurement of pH and by visual inspection. Rheological properties were studied using the cone and plate method at shear rates of 600 to 6000 1/s. Viscosities corresponding to shear rates were also calculated. Our results indicated that C2001 could be used as a gelling agent for IND in topical preparations. IND-C2001 gels were clear and exhibited an acceptable appearance; gel behaviour was non-Newtonian and pseudoplastic. The addition of either Tween 80 or PVP 25 to the base gel formulation significantly increased the shear stresses and viscosity of the gels. These novel formulations exhibited good physical stability throughout the 4-week examination periods as inferred from pH measurements and microscopic examination. Additionally, the non-Newtonian pseudoplastic behaviour of the gels was maintained throughout storage, with only a minimal decrease in gel viscosity after 4 weeks. Differences in consistencies of the four formulations, although initially apparent, were no longer evident after 4 weeks of storage for all temperature conditions examined. Gels stored at high temperature (45 degrees C) developed a dark yellow color and decreased in viscosity compared to other storage temperature. PMID- 12641333 TI - Characterization and pharmacological actions of tecostanine, an alkaloid of Tecoma stans. AB - Tecostanine (1) was isolated from Tecoma stans leaves. Its sterochemistry was elucidated as well as its antihyperglycemic activity and its affinity to opioid and nicotinic receptors. The oxalate salt of 1 did not significantly affect blood glucose levels in normoglycaemic and hyperglycaemic rats. It did not appear to interact with opioid receptors (mu type) and showed only moderate affinity to the nicotinic receptor. PMID- 12641332 TI - A toxicological evaluation of 2-dimethylaminomethyl-1-phenyl-2-propen-1-one hydrochloride. AB - 2-Dimethylaminomethyl-1-phenyl-2-propen-1-one hydrochloride (3) is a novel cytotoxic and anticancer agent. The objective of this study was to obtain information pertaining to possible toxic symptoms detected by in vivo evaluations in mice and an in vitro test for mutagenicity. The data obtained revealed that 3 had no effect on alanine transaminase, aspartate transaminase, HDL cholesterol and protein concentrations in sera nor were variations in the numbers of red and white blood cells detected. Furthermore autopsies of treated mice revealed no pathological symptoms in the heart, kidney, brain, spleen and testes. However elevation of the concentrations of total cholesterol, triglycerides, creatinine and urea were noted in treated mice as well as inflammation of the liver and lungs. Chromosomal aberrations were detected in a micronuclei test. In the Ames test, compound 3 was converted into one or more mutagens in the presence (but not the absence) of a murine liver homogenate. Thus future molecular modifications of 3 should bear in mind approaches to reduce or minimize unwanted side effects. PMID- 12641334 TI - Bioactive kaurane diterpenes and coumarins from Fortunella margarita. AB - Two bioactive pyranocoumarins 1 (sesselin) and 3 (xanthyletin) and one prenylated coumarin 2 (suberosin), beside three rare kaurene diterpenes 5-7 were isolated from the roots of Fortunella margarita. Their structures were determined from their spectroscopic data, including 1H/13C 2D NMR experiments. The kaurene diterpenes 5-7 are reported in Rutaceae for the first time. Diterpene 5 was found to be a potent stimulator of uterine contraction; it also caused stimulation of brain activity. PMID- 12641336 TI - The history of electrotherapy of pain--or: what Voltaren has to do with voltage. AB - To reasons not yet completely understood, the pharmaceutical trademark Voltaren or Voltarol refers to the Italian scientist Alessandro Volta (1745-1827) who played an important role in the history of electricity. It is shown that electrotherapy has been used for the treatment of pain, gout and rheumatic disorders from ancient times to the present day. This therapeutic attempt was also discussed in scientific literature in the late 1960s, the time Voltaren was under development. Therefore, it is suggested that the electrotherapy of pain in history is a background for chosing this trademark. PMID- 12641335 TI - Cytotoxic triterpenes from Ligulariopsis shichuana. AB - Five olean-12-ene triterpenes (1-5) were isolated from the whole plant of Ligulariopsis shichuana. Their structures were elucidated by spectroscopic methods, including IR, EIMS positive HRSIMS, 1DNMR, and 2DNMR. Among them, 16beta,28-dihydroxyolean-12-en-3-one (3), olean-12-en-3beta,6beta,16beta,28 tetraol (4), 6beta,16beta-dihydroxyolean-12-en-3-one (5) are new compounds. In addition, compounds 4 and 5 showed cytotoxic activities on human hepatomacells (SMMC-7721), human ovarian neoplasm cells (HO-8910) and human hepatocytes cells (LO2). PMID- 12641337 TI - Loratadine: hydroxymethylation in syrup. PMID- 12641338 TI - In vitro release studies on topical gel formulations of nimesulide. PMID- 12641339 TI - Study of local anaesthetics, part 161: Influence of additives upon the release of a potential drug of a group of esters of phenylcarbamic acids from a hydroxyethyl cellulose hydrogel. PMID- 12641340 TI - A new flavonol glycoside from leaves of Zizyphus lotus. PMID- 12641341 TI - Dendritic cells under investigation in autoimmune disease. AB - Autoimmune disorders play an increasing role in public health, especially in light of the fact of the growing aged population, which primarily develop such diseases. A clear understanding of the mechanisms leading to the development of autoimmune responses and finally to autoimmune disease does not exist. Autoimmunity is characterized by the presence of autoantibodies and/or autoreactive T cells and the corresponding organ manifestation. Following the discovery of autoreactive T cells found in the periphery of mice and humans, the old immunological concept that autoreactive T cells are completely deleted in the thymus during evolution has been revised in recent years. Although antigen presenting cells and particularly dendritic cells are known to play an important role in the regulation of immune responses and the activation of T cells, recent evidence suggests that the role of dendritic cells in the development of autoimmunity has been underestimated previously. This article aims to give a general overview on the basic immunological principles involved and gives a short review of the current literature on the functional relevance of dendritic cells in various human and murine autoimmune disorders. PMID- 12641342 TI - Biogenic amines and polyamines: similar biochemistry for different physiological missions and biomedical applications. AB - Biogenic amines are organic polycations derived from aromatic or cationic amino acids. All of them have one or more positive charges and a hydrophobic skeleton. Nature has evolved these molecules to play different physiological roles in mammals, but maintains similar patterns for their metabolic and intracellular handling. As deduced from this review, many questions still remain to be solved around their biochemistry and molecular biology, blocking our aims to control the relevant pathologies in which they are involved (cancer and immunological, neurological, and gastrointestinal diseases). Advances in this knowledge are dispersed among groups working on different biomedical areas. In these pages, we put together the most relevant information to remark how fruitful it can be to learn from Nature and to take advantage of the biochemical similarities (key protein structures and their regulation data on metabolic interplays and binding properties) to generate new hypothesis and develop different biomedical strategies based on biochemistry and molecular biology of these compounds. PMID- 12641343 TI - Iron metabolism in the reticuloendothelial system. AB - Comprised mainly of monocytes and tissue macrophages, the reticuloendothelial system (RES) plays two major roles in iron metabolism: it recycles iron from senescent red blood cells and it serves as a large storage depot for excess iron. Although iron recycling by the RES represents the largest pathway of iron efflux in the body, the precise mechanisms involved have remained elusive. However, studies characterizing the function and regulation of Nramp1, DMT1, HFE, FPN1, CD163, and hepcidin are rapidly expanding our knowledge of the molecular aspects of RE iron handling. This review summarizes fundamental physiological and biochemical aspects of iron metabolism in the RES and focuses on how recent studies have advanced our understanding of these areas. Also discussed are novel insights into the molecular mechanisms contributing to the abnormal RE iron metabolism characteristic of hereditary hemochromatosis and the anemia of chronic disease. PMID- 12641344 TI - A novel approach to the treatment of gunshot injuries to the sacrum. AB - Two patients (a 17-year-old male and a 19-year-old female) sustained single gunshot wounds to the abdomen. Given the nature of their wounds and presumed peritoneal penetration they were taken urgently to the operating room for exploratory laparotomy. Both patients sustained multiple intra-abdominal injuries including gastric and small and large bowel perforations. Given the extensive nature of these injuries both patients required bowel resections. Upon further exploration they were both noted to have significant bleeding from the bony aspect of their sacrum. A surgical hemostat (CoStasis; Cohesion Technologies, Inc., Palo Alto, CA) (4.5-9.0 cm3) was directly applied to the injured area of the sacrum in both cases after standard surgical hemostatic techniques were unsuccessful. After application no further attempts of hemostatic control by standard methods were attempted. In both cases the sacral bleeding ceased without any further surgical intervention. Postoperatively neither patient rebled from the sacral injuries. Penetrating injuries to the sacrum can be life threatening and difficult to control with standard surgical hemostatic techniques. CoStasis, a new surgical hemostat, was effective in obtaining immediate and definitive hemostatic control. Future prospective trials on the use of CoStasis in trauma patients are warranted. PMID- 12641345 TI - A study of the patient factors affecting reconstruction after mastectomy for breast carcinoma. AB - Breast shape may be retained after surgery for breast cancer with either wide local excision (WLE) and radiotherapy or mastectomy (M) and immediate reconstruction (M+R). We determined the proportions of patients who would be suitable for these options, would accept them, and if they declined M+R their reasons for doing so. Over a 10-month period 177 women were assessed at a combined breast clinic by general and plastic surgeons and by radiation oncologists. A prospective record was made of the patient demographic data, the clinical decisions (and their reasons), and the patient choices (and their reasons). A transverse rectus abdominis myocutaneous flap was the commonest method of reconstruction. One hundred thirty-five (76%) were judged to be suitable for locoregional surgery. Of these M+R was offered to 83 patients, whereas 53 were not considered because of combinations of cosmetic considerations (31), risk factors (25), old age (13), and oncological factors (nine). Fifty-one of the 83 (61%) offered M+R declined it because they preferred a simpler procedure (34), regarded breast appearance as unimportant (15), preferred breast conservation (five), did not have a partner (three), felt that they were too old (two), or had religious reasons (two). Ultimately 69 (51%) underwent M, 34 (25%) WLE, and 32 (24%) M+R. There was no correlation between acceptance or not of M+R and age, race, employment, education level, or marital status. We conclude that many patients were suitable for M+R, but fewer than half accepted it; this decision was unrelated to age, race, employment, or marital status. PMID- 12641346 TI - Carcinoid tumor of the biliary tract: treating a rare cause of bile duct obstruction. AB - Biliary carcinoids are rare with fewer than 30 cases reported in the English literature. The objective of this report is to describe an additional patient found to have a biliary carcinoid and to define the presentation, diagnosis, and management of patients with this rare biliary tumor. In our case the patient initially presented with clinical jaundice and elevated transaminases. Endoscopic retrograde cholangiogram established a mass suspicious for cholangiocarcinoma (Klatskin tumor). The patient was initially managed with an endostent, which was later removed in favor of a percutaneous transhepatic cholangiogram tube. At the time of surgery successful removal of a firm nodular mass at the area of the ductal bifurcation was achieved and biliary continuity re-established with a Roux en-Y hepaticojejunostomy. Pathology revealed carcinoid tumor of the bile duct with one lymph node positive for tumor. The patient did not receive any adjuvant radiation or chemotherapy. This case serves to highlight that extrahepatic biliary carcinoids constitute a rare but identifiable subset of bile duct tumors. Diagnostic workup should include US, CT, and cholangiography. Surgical exploration is universally indicated in physiologically fit patients with operative management to include resection and re-establishment of biliary continuity. Data on adjuvant therapy remain investigational; however, available information suggests that patients with biliary carcinoid have an overall favorable prognosis after aggressive surgical management. PMID- 12641347 TI - Serial abdominal closure technique (the "SAC" procedure): a novel method for delayed closure of the abdominal wall. AB - Abdominal compartment syndrome may occur after any elective or emergent abdominal operations that are complicated by postoperative hemorrhage or in the trauma patient who has massive fluid replacement for intra-abdominal bleeding. Once the abdomen is decompressed the type of closure varies as much as the surgeon performing the procedure. We have devised a simple, reproducible, inexpensive, and safe method to close the abdomen at the bedside. Serial abdominal closure (SAC) was performed on three patients 45, 54, and 14 years of age who had developed abdominal compartment syndrome secondary to an upper gastrointestinal bleed requiring massive transfusion, a tear of the superior mesenteric vein, and a grade 4 liver laceration respectively, all necessitating abdominal decompression. All three patients had their abdominal wounds closed at the bedside over the course of several days with our SAC technique. Subsequent postoperative course was uneventful and the abdominal wall was free of defects at one-year follow-up. SAC is an efficient, inexpensive, and easily reproducible method of managing the open abdomen. The use of SAC prevented abdominal closure related complications such as enteric fistula and hernia formation in our three patients. PMID- 12641348 TI - Splenic rupture from metastatic granulosa cell tumor 29 years after curative resection: case report and review of the literature. AB - Granulosa cell tumors account for 2 to 3 per cent of all ovarian neoplasms. This case report describes a patient with splenic rupture from a bulky metastatic deposit 29 years after her initial surgery. A review of the literature regarding the patterns and timing of recurrence is also discussed. PMID- 12641349 TI - Recurrent spigelian hernia: a case report. AB - Only seven cases of spigelian hernia recurrence have been previously reported. We report the case of a 75-year-old male patient who presented with extremely large hernia after four unsuccessful suture repairs over 12 years. The abdominal wall defect was repaired with Marlex mesh. The advantage of using prosthetic mesh in both primary and recurrent spigelian hernia is supported by recent clinical research data indicating a generalized collagen metabolism disorder in patients with primary and recurrent hernia. Mesh repair allows for tension-free anatomic restoration of distorted tissues associated with repair failures. PMID- 12641350 TI - Technical limitations of sentinel node biopsy in breast cancer: a single surgeon's experience. AB - Few studies have attempted to critically identify patient- and tumor-related factors that limit sentinel node biopsy (SNB). These studies have been limited by sample size and surgeon variability. The present study attempts to enumerate these limitations in a unique group of patients. One hundred twenty-five SNBs performed by a single surgeon between May 1997 and June 2001 were reviewed. Overall SNB was successful in 96 per cent of patients with a 97 per cent correlation with the axillary node dissection. Sentinel node identification was not affected by age, tumor size, tumor location, prior segmental resection, or neoadjuvant therapy. No false negatives were noted in the neoadjuvant group. The use of blue dye alone significantly understaged patients when compared with isotope alone (P = 0.02). SNB is a highly accurate method to identify axillary metastases and its limitations are not affected by patient or tumor related factors. In the present study SNB detection by both isotope and blue dye has been shown to be superior to blue dye alone. This finding demonstrates that these limitations may be overcome with the standardization of the technique used. PMID- 12641351 TI - The role of [18F]fluorodeoxyglucose positron emission tomography imaging in the evaluation of hepatocellular carcinoma. AB - It has been well established that hepatocellular carcinomas (HCCs) accumulate [18F]fluorodeoxyglucose (FDG) to varying degrees; this is thought to be due to differing amounts of FDG-6-phosphatase activity. The purpose of this study was to evaluate the impact of FDG imaging on the management of patients diagnosed with hepatocellular carcinoma. We conducted a retrospective review of the clinical data of 91 consecutive patients diagnosed with HCC who underwent FDG-positron emission tomography (PET) imaging between August 1993 and March 2001. The patients were divided into two groups. In Group one 67 of 91 (74%) patients were evaluated for proven but untreated hepatic lesions using PET. In Group two the remaining 24 patients (26%) were referred for evaluation of HCC recurrence but did not have prior PET. The FDG images were acquired with two dedicated PET tomographs [Siemens ECAT 933, CTI (Knoxville, TN) and GE Advance, General Electric Medical Systems (Milwaukee, WI)] one hour after the intravenous administration of 10 mCi of FDG. Tumor biopsy or resection specimens were available for review from 34 patients and were evaluated for histologic grade, presence of cirrhosis, tumor necrosis, and intratumoral fibrosis. In group one 43 of 67 (64%) of the HCCs accumulated FDG. Sixteen of the 43 patients in whom FDG was accumulated had multiple subsequent FDG-PET scans either for monitoring therapy or for detection of recurrence. FDG-PET imaging had an impact on the management of 20 of these patients: by guiding the biopsy at the metabolically active site of a large necrotic tumor (one), by identifying distant metastases (five), by monitoring the response to treatment with hepatic chemoembolization and guiding additional regional therapy (12), and by detecting recurrence (two). In group two recurrence and/or metastases were demonstrated with FDG-PET imaging in six of 24 (25%) patients, three of whom had multiple subsequent FDG-PET scans to monitor their treatment. Higher histopathologic grade and intratumoral fibrosis but not necrosis or cirrhosis correlated with PET positivity. In this study only 64 per cent of HCCs accumulated FDG. Despite this limitation FDG-PET imaging remains a useful tool in the diagnosis and treatment of HCC. FDG-PET imaging had a clinically significant impact in 26 of 91 (28%) patients with HCC. This includes detection of unsuspected metastatic disease in high-risk patients including liver transplant candidates-and monitoring response to hepatic-directed therapy. FDG-PET should be considered as part of the workup and management of selected patients with HCC. PMID- 12641352 TI - Low-dose adjuvant interferon for stage III malignant melanoma. AB - The role of interferon as adjuvant therapy in stage III melanoma has recently been questioned. Prospective randomized studies have shown conflicting results regarding the efficacy of adjuvant treatment. The purpose of this study was to examine the use of low-dose adjuvant interferon alpha2-b (IFN) in stage III melanoma patients treated at a single institution. This study was a retrospective analysis of 60 stage III melanoma cases from Wake Forest University treated between 1983 and 1998. Cases were identified via the tumor registry. All patients underwent standard lymphadenectomy after diagnosis. After recovery from surgery patients were offered low-dose IFN (3 million units subcutaneous TIW for 1 month and then 6 million units subcutaneous TIW for 11 months) as adjuvant therapy for stage III melanoma. The patients were followed up jointly by medical and surgical oncology. There were 39 male and 21 female patients with mean age of 60.0 (range 37-89) years. The average number of positive nodes was 3.6 (median = 1) for the treated group and 1.8 (median = 1) for those untreated (P = 0.71). The average tumor thickness was similar between groups: 4.71 versus 4.88 mm for the IFN and observation groups respectively. The IFN group (N = 25) that received low-dose treatment had a median survival of 7.9 years with a 5-year survival rate of 69 per cent. The 35 cases that did not receive interferon had a median survival of 6.5 years and a 5-year survival rate of 52 per cent. These survival rates were not significantly different (P = 0.91). The median disease-free survival for patients who did not receive IFN treatment was 2.4 years and 1.4 years for the treated group (P = 0.19). The data show that there was similar survival for those who did and did not receive the low-dose IFN treatment. Although only large prospective trials can conclusively exclude a small survival time this experience suggests that there is no significant survival advantage to low-dose adjuvant IFN therapy for stage III melanoma patients. Hopefully upcoming cooperative group trials will clarify the potential value of adjuvant IFN in this setting. However, although the toxicity of this regimen was mild we suggest that low-dose adjuvant IFN for stage III melanoma should not be utilized outside the setting of a clinical trial. PMID- 12641353 TI - Bacterial lipopolysaccharide-stimulated release of tumor necrosis factor-alpha from the isolated rat heart: the effect of aprotinin and forskolin. AB - Aprotinin has been reported to reduce plasma levels of inflammatory cytokines associated with cardiopulmonary bypass (CPB). Because CPB is also associated with elevated levels of bacterial lipopolysaccharide (LPS) and LPS stimulates release of inflammatory cytokines from the heart we tested the hypothesis that aprotinin would inhibit cardiac release of tumor necrosis factor-alpha (TNF) provoked by LPS. Isolated rat hearts were perfused Langendorf style. After 30 minutes of equilibration LPS (100 ng/mL) was infused for 60 minutes. Timed samples of coronary effluent were collected at 0, 30, 60, 90, 120, and 150 minutes after the initiation of LPS for the measurement of coronary flow and the determination of TNF and cyclic AMP. Other hearts were perfused with buffer containing aprotinin [137 kallikrein-inhibiting units (KIU)/mL or 250 KIU/mL] and then infused with LPS. An additional group received forskolin (10 microM) and LPS. In hearts perfused as controls with buffer alone no TNF was detected in the coronary effluent. In hearts perfused with LPS TNF was reliably detected in the coronary effluent at 60 minutes (606 +/- 450 pg/min) and increased with time to a level of 1792 +/- 650 pg/min at 150 minutes. The addition of aprotinin had no significant effect on LPS-stimulated TNF release. For instance in hearts perfused with 137 KIU/mL aprotinin LPS-stimulated release at 150 minutes was 2141 +/- 732 pg/min and in hearts perfused with 250 KIU/mL LPS-stimulated TNF release was 2049 +/- 789 pg/min. Forskolin administration was associated with release of cyclic AMP from the heart and completely inhibited LPS-stimulated TNF release. We conclude that LPS stimulated release of TNF from the heart. Adding aprotinin to the perfusion buffer in either high or low concentrations did not attenuate LPS stimulated cytokine release. Elevating myocardial cyclic AMP with forskolin completely attenuated LPS-stimulated TNF release. PMID- 12641354 TI - Synchronous occurrence of glioblastoma multiforme and esophageal adenocarcinoma. AB - Synchronous malignancies are rare occurrences for which there may be a genetic link between two cancers or which may be simply coincidental. Although glioblastoma multiforme and esophageal adenocarcinoma have few clinical similarities there are no known biochemical or genetic links between the two malignancies. This case discussion details the synchronous occurrences of these two lesions and highlights possible clinical, biochemical, and genetic commonalities. PMID- 12641355 TI - Physiologic predictors of survival in post-traumatic arrest. AB - Traumatic cardiac or pulmonary arrest is often associated with a dismal outcome and is considered by many to be an example of medical futility and inappropriate use of resources. This study aimed to identify the predictors of survival in patients experiencing traumatic cardiac arrest. We retrospectively reviewed all trauma patients undergoing cardiopulmonary resuscitation on arrival to the Emergency Department (ED) at an American College of Surgeons-designated Level I trauma center over 4 years. ED survival, hospital survival, and neurologic outcomes on discharge were the primary outcomes. Survival rates were examined in relation to demographics, mechanism of injury, airway management, cardiac electrical rhythm, and pupil size and reactivity. Statistical analyses used chi square and t tests, P < 0.05 was considered significant. A total of 195 patients arrived in the ED with traumatic cardiac arrest; 34 were pronounced dead on arrival (no signs of life), and no resuscitation efforts were initiated. Of the remaining 161 patients 53 (33%) survived to leave the ED, and only 15 (9%) left the hospital alive. Demographic features were similar in survivors and nonsurvivors. The setting of intubation (prehospital vs ED) did not influence survival (P = 0.36). Penetrating trauma adversely affected survival in the ED (P = 0.01); however, this only approached significance in the final outcome of hospital survival (P = 0.06). The presence of sinus rhythm and nondilated reactive pupils was highly significant in predicting ED and hospital survival (P = 0.001). No patient with agonal rhythm or ventricular fibrillation/tachycardia survived, and 14 of the 15 hospital survivors had reactive pupils on arrival to the ED. We conclude that sinus rhythm and pupil size and reactivity are important physiologic variables that predict potential survival and may be used to guide continuation of resuscitative efforts in patients with traumatic cardiac arrest. PMID- 12641357 TI - Trends in preparation for colorectal surgery: survey of the members of the American Society of Colon and Rectal Surgeons. AB - The utility of antibiotic and mechanical preparation for colorectal surgery is controversial, and numerous different regimens are used. The aim of this study was to detect trends in preparation for surgery among American colon and rectal surgeons. Members of the American Society of Colon and Rectal Surgeons practicing in the United States were surveyed with a postal questionnaire regarding their routine preparations for colon and rectal surgery. Five hundred fifteen (40%) of the 1295 questionnaires sent were returned. Eighty-one per cent of the respondents had completed an accredited colorectal training program, and the average experience in practice was 13.7 (+/- 8.7) years. Half of the surgeons felt that prophylactic oral antibiotic is essential, 41 per cent felt it was doubtful, and 10 per cent considered oral prophylaxis unnecessary. Despite these statements 75 per cent of the surgeons routinely utilized oral antibiotics (96% of them used a combination of two drugs), 11 per cent used them selectively, and only 13 per cent omitted oral prophylaxis from their practice. Similarly although the usefulness of intravenous antibiotics was questioned by 11 per cent of the surgeons 98 per cent routinely used them. The average number of postoperative doses was two (+/- 1.9). Although 10 per cent of the surgeons questioned the importance of mechanical preparation more than 99 per cent routinely used it. Forty-seven per cent of the surgeons used sodium phosphate, 32 per cent used polyethylene glycol, and 14 per cent alternated between these two options. We conclude that although the use of oral antibiotic prophylaxis for colorectal surgery is controversial among surgeons it is still routinely practiced by 75 per cent. Intravenous antibiotic prophylaxis and mechanical cleansing, however, are still a dogma and almost invariably used. There is a trend toward the use of a shorter course of postoperative intravenous antibiotics and the use of sodium phosphate for mechanical cleansing. PMID- 12641356 TI - Acute lower gastroenteric bleeding retrospective analysis (the ALGEBRA study): an analysis of the triage, management and outcomes of patients with acute lower gastrointestinal bleeding. AB - Many algorithms have been developed for patients with acute lower gastrointestinal hemorrhage (ALGIH). Their clinical usefulness is not readily apparent. It is important first to observe patterns in admission, triage, and management to formulate hypotheses as to how outcomes might be affected. We reviewed patient charts with the diagnosis of gastrointestinal hemorrhage from June 1998 to January 2001. Patients with ALGIH were entered into a database. We defined patients as having ALGIH if presentation included melena or hematochezia. Patients with hematemesis, bloody nasogastric aspirate, or occult fecal blood were excluded. Observations were made on 420 patients. Seventy-six per cent of patients were admitted to the medical service. Lower endoscopy was the first diagnostic method in 33 per cent. Medical management comprised 52 per cent of first management strategies. Surgeons used angiography (3% vs 1%) or surgery (25% vs 5%) more than other services. Fourteen per cent of patients managed with endoscopy, 16 per cent medically, 17 per cent with surgery, and 67 per cent with interventional radiology required two or more subsequent packed red blood cell transfusions. Mean admission Acute Physiology and Chronic Health Evaluation II score was 9.2 whereas that for those with mortality was 13.5. We conclude that the construction of a database will allow for formation and testing of hypotheses in managing ALGIH. PMID- 12641358 TI - Endovascular repair of acute innominate artery injury due to blunt trauma. AB - The utility of endovascular techniques has expanded greatly over the past decade. Physicians now have choices regarding the treatment of many injuries that have traditionally required open surgical repair. Technological advances in materials as well as improved training and expertise among practitioners has led to increased availability of endovascular procedures that can often provide an effective and less invasive means of management. The following case report describes the successful treatment of a traumatic blunt injury to the innominate artery using endovascular techniques. Also provided is a review of the physical and radiographic findings associated with innominate artery rupture as well as a discussion on the diagnosis and treatment of such an injury. Isolated injuries of the innominate artery are exceedingly rare, and very little has been published about the endovascular repair of this specific injury. An extensive MEDLINE search was conducted to investigate whether or not endovascular repair of this particular injury had yet been described, and we found no published reports in the American medical literature. Although the technical aspects of this case are not particularly unique this case report demonstrates yet another successful application of endovascular intervention in the acute setting of blunt injury. PMID- 12641359 TI - Laparoscopic repair of a recurrent chronic traumatic diaphragmatic hernia. AB - Traditionally the approach to a long-standing traumatic diaphragmatic hernia has been a closure of the diaphragmatic defect via a thoracotomy. The evolution of minimally invasive surgery has allowed surgeons to challenge many of the traditional approaches. Herein we describe the first reported case of laparoscopic repair of recurrent chronic traumatic diaphragmatic hernia as well as review the current literature on minimally invasive surgery for traumatic diaphragmatic hernia. With proper advanced laparoscopic skills and techniques laparoscopic diaphragmatic herniorrhaphy for a chronic and recurrent defect is a safe and viable option. PMID- 12641360 TI - Liposarcoma of the spermatic cord masquerading as an incarcerated inguinal hernia. AB - We present a rare case of liposarcoma of the spermatic cord. There are only 61 reports in the literature. The presenting complaint is usually a painless bulge in the inguinal or scrotal region. Our patient presented with a new-onset inguinoscrotal swelling that was misdiagnosed preoperatively as an incarcerated indirect hernia. The treatment for a spermatic cord liposarcoma is radical orchiectomy with high ligation of the cord. Radiation therapy is recommended in addition to surgery in situations with evidence of tumor with propensity for more aggressive behavior (i.e., high-grade tumor, lymphatic invasion, inadequate margin, or recurrence). The current literature, diagnosis, and management of malignant tumors of the spermatic cord are reviewed. PMID- 12641361 TI - Perianal mucinous adenocarcinoma: unusual case presentations and review of the literature. AB - Perianal mucinous adenocarcinoma is a rare cancer constituting 3 to 11 per cent of all anal carcinomas. It may arise de novo or from a fistula or abscess cavity. We present two cases of this disease process. Case One is a 52-year-old man with a chronic history of perianal abscesses who presented to the emergency room with a large bowel obstruction. He required diversion and wide local excision with lateral internal sphincterotomy for relief of the obstruction. Pathology from the excised material revealed the unexpected diagnosis of invasive mucinous adenocarcinoma of the anus. Case Two is a 59-year-old man with a chronic history of complex fistulas and abscesses who presented to our office with a horseshoe fistula and deep postanal space abscess. Because of the nonhealing nature of the wound, biopsies from the abscess crater, fistulous tract, and the perianal skin opening were taken. The pathology department identified the specimens as invasive mucinous adenocarcinoma of the anal canal. This is an aggressive cancer often misdiagnosed clinically as benign pathology. A high index of suspicion and biopsy of fistulous tracts and abscesses are the keys to early diagnosis and treatment. With combination chemotherapy and radiation therapy in conjunction with aggressive surgical resection long-term survival might be obtained. PMID- 12641362 TI - Limitations of technetium 99m sestamibi scintigraphic localization for primary hyperparathyroidism associated with multiglandular disease. AB - Successful surgical treatment of primary hyperparathyroidism requires the localization and excision of the parathyroid tissue responsible for excessive parathyroid hormone secretion while ensuring that the patient will have sufficient endogenous parathyroid hormone production to maintain eucalcemia. In selecting patients with primary hyperparathyroidism for unilateral parathyroidectomy the surgeon should be able to diagnose multiglandular disease either preoperatively or intraoperatively. We performed a retrospective review of 123 patients who underwent surgical treatment for primary hyperparathyroidism to determine the potential feasibility of selecting patients for minimally invasive surgery based on preoperative imaging studies. All patients were studied preoperatively with 99m technetium-sestamibi scintigraphy. High-resolution ultrasonography was performed in 119 of these patients. All patients except one underwent bilateral cervical exploration. A patient with an intrathoracic adenoma was successfully diagnosed by scintigraphy thereby allowing treatment by a limited thoracotomy. One hundred eight patients had solitary adenomas and 15 had multiglandular disease. In none of the patients with bilateral multiglandular disease were all abnormal glands localized preoperatively. Patients in our study with primary hyperparathyroidism and multiglandular disease were underdiagnosed by preoperative imaging. A minimally invasive approach based solely on preoperative imaging studies may result in treatment failure in patients with multiglandular involvement. PMID- 12641363 TI - Dexon mesh splenorrhaphy for intraoperative splenic injuries. AB - The preferred management option for intraoperative splenic injuries is organ repair and preservation rather than splenectomy given the important immunologic function of the spleen. Wrapping the injured spleen with a Dexon mesh has been shown to be an effective alternative to splenectomy for significant splenic bleeding. However, this technique uses a foreign body that carries a theoretical infectious risk particularly in cases in which the alimentary tract has been opened. This study was undertaken to evaluate whether Dexon mesh splenorrhaphy when used for intraoperative splenic injuries was associated with significant infectious complications. The clinical courses of 23 patients who had Dexon mesh splenorrhaphy performed at a university teaching hospital for intraoperative splenic injury from 1991 to 1999 were reviewed. Eleven patients (48%) had their gastrointestinal tract opened during the surgery. No patients developed an intra abdominal abscess or required reoperation for bleeding. The most common postoperative complications were left lower lobe atelectasis (18 patients, 78%), postoperative fever (13 patients, 56%), and left pleural effusion (12 patients, 52%). Dexon mesh splenorrhaphy effectively controls splenic bleeding due to intraoperative injury without significant infectious complications. PMID- 12641364 TI - Is the Socratic method illegal? PMID- 12641365 TI - Association of depression with agitation in elderly nursing home residents. AB - Agitation is a serious problem for elderly individuals with dementia. It is often the major reason for admission to a restrictive environment such as a nursing home or hospital. The objectives of the current study were to (1) identify the components of agitation embedded in the Psychogeriatric Dependency Rating Scale (PGDRS) and (2) find race, gender, depression, and cognitive deficits associated with the factors extracted from the PGDRS in demographic variables and clinical variables. The study sample comprised 2285 subjects who were admitted to 59 nursing homes across Maryland. The factor analysis of the PGDRS confirmed that agitation is made up of a number of different elements ranging from physical and/or verbal aggression to wandering. Correlates of these elements varied, as did possible treatments. For example, physical and/or verbal aggression often accompanied severe depression, suggesting that treating the depression may alleviate this problem. However, wandering and psychotic behavior may be less amenable to existing treatments as these behaviors were associated with severe cognitive impairment. PMID- 12641366 TI - Correlates of behavioral disturbances and pattern of psychotropic medication use in five skilled nursing facilities. AB - There are several treatment options for behavioral disturbances (BDs) in dementia. However, the choice of a specific psychotropic agent is directed by personal preferences and local community practice patterns. We examined the relationship between common clusters of BDs and the use of different classes of psychotropic agents in our community. A cross-sectional study of 430 long-term care residents from 5 nursing homes was undertaken. The Behavior Measurement Scale (BMS) was used to measure the frequency of BDs grouped in 4 categories. Residents with > 4 BD episodes in at least one category during a 2-week observation period were the behavior group and were considered to have clinically significant BDs. A sample of patients who had < 4 BDs in all BMS categories during the same observation period defined the nonbehavior group. A BD cluster was defined as > 4 BDs occurring in one or more BMS categories during the 2-week observation. Data on functional status, comorbidity, use of benzodiazepines, antidepressants, and neuroleptic agents were collected with chart review. The chi square test was used to examine the correlation between variables. Clinically significant BDs were identified in 27.2% (117/430) of the residents in the sample. Five of 15 behavior clusters accounted for 73% of all clinically significant BDs. The 5 clusters were verbally nonaggressive behaviors (cluster 1, 20.5%), behaviors from all 4 categories (cluster 2, 17.9%), verbally and physically nonaggressive behaviors (cluster 3, 14.5%), physically nonaggressive behaviors (cluster 4, 12.8%), and verbally aggressive and nonaggressive behaviors (cluster 5, 7.7%). Cluster 5 had a negative correlation with functional impairment (P = .009). There was a significant correlation between cluster 2 and benzodiazepine use (P = .014). No other significant correlation was found between any of the 5 clusters and demographic variables, comorbidity status, and use of antidepressant or neuroleptic medications. Residents in the behavior group had higher impairment in self-feeding (P = .036) and bathing (P < .001) and were more likely to be treated with benzodiazepines (P = .004) and neuroleptic agents (P = .009) than residents in the nonbehavior group (n = 116). The higher use of neuroleptics and benzodiazepines in the behavior group compared with the nonbehavior group indicates that BDs are being identified for treatment, but the medications used may not be efficacious. The lack of association between specific classes of psychotropic medications and distinct behavior clusters indicates that clinicians are not using a standardized approach to target the neurochemical abnormalities that may underlie certain behavior clusters. Some behavior clusters correlate with impairment in specific activities of daily living categories such as bathing and feeding, making room for nonpharmacologic interventions. PMID- 12641367 TI - Health service utilization by Alzheimer's disease patients: a 2-year follow-up of primary versus subspecialty care. AB - All dementia patients and their caregivers who had received a university-based comprehensive evaluation and a diagnosis of Alzheimer's disease during 1997 (N = 80) were identified. Of the original cohort, 48.8% (n = 39) were able to be contacted approximately 2 years after their initial assessment, and the caregivers were the informants for this follow-up. Two subgroups were defined: 22 patients were being seen only by their primary care physicians (MED), while 17 patients were also being treated by a geriatric psychiatry faculty member (GERO). There were statistically significant differences between the 2 groups (MED versus GERO, respectively) at follow-up in terms of (1) institutionalization (30.0% versus 4.6%, P < .05), (2) CDR (2.3 versus 1.5, P < .005), and (3) prescription of donepezil at follow-up (45.5% versus 76.5%, P = .05). These differences are being assessed in a larger scale prospective study. PMID- 12641368 TI - Smoking and cognitive performance in the community elderly: a longitudinal study. AB - This prospective study investigated the association between smoking and cognitive performance in a community of nondemented elderly subjects aged 65 or older. All subjects were categorized as current smokers, former smokers, or never smokers. The lifetime cigarette exposure was computed. At baseline, we found the abstainers from smoking had better cognitive performances; however, the differences were not significant after adjusting for age, education, hypertension, diabetes, and vascular events. The lifetime cigarette exposure was not predictive of the cognitive status. At a 3-year follow-up, neither the smoking status nor the lifetime cigarette exposure predicted the declination of cognition. PMID- 12641369 TI - Construct validity of the 15-item geriatric depression scale in older medical inpatients. AB - The construct validity of the 15-item Geriatric Depression Scale (sfGDS) has been assessed in selected populations. The aim of this study was to assess the appropriateness of applying the sfGDS to unselected older inpatients. The main component analysis of sfGDS was performed in 2032 medical inpatients (mean age = 76.3 +/- 8.4). sfGDS did not qualify as a unidimensional test. Three factors explained 47.7% of variance and explored the following dimensions: positive attitude toward life, distressing thoughts/negative judgment about the own condition, and inactivity/reduced self-esteem. The internal homogeneity was poor (Cronbach's alpha = .46). A higher fraction of variance was explained in patients independent in all or dependent in > or = 1 activity of daily living (ADL). In older medical inpatients, sfGDS is not a single construct, which prevents the univocal interpretation of the final score. The higher fraction of explained variance in patients with comparable ADL performance probably reflects the dependency of affective from physical status. PMID- 12641370 TI - Treatment of delirium in older adults with quetiapine. AB - Delirium is a neuropsychiatric syndrome characterized by impairment of consciousness, changes in cognition, or perceptual disturbances. In addition, delirium is often accompanied by delusions, hallucinations, and agitation. In this study, 12 older patients with delirium were treated for neuropsychiatric symptoms with quetiapine. The mean duration for stabilization was 5.91 +/- 2.22 days, and the mean dose was 93.75 +/- 23.31 mg/day. None of the 12 patients developed extrapyramidal symptoms. There were significant improvements on all measures used in this study. Interestingly, the Delirium Rating Scale scores along with scores of the Mini-Mental State Examination and Clock Drawing Test continued to improve throughout the 3-month study period. In our study, we found that quetiapine was a safe and effective treatment in hospitalized older patients with delirium. PMID- 12641371 TI - Recorded delirium in a national sample of elderly inpatients: potential implications for recognition. AB - This retrospective study examined delirium and related confusional diagnoses recorded in patients older than age 60 discharged from Veterans Affairs (VA) acute inpatient units nationally in 1996 (n = 267,947). Only 4% of patients had delirium or related confusional diagnoses recorded. Patients with recorded delirium had significantly higher mortality than did those without recorded delirium or those with other confusional diagnoses ("organic psychoses"); the most common delirium types were dementia with delirium and alcohol intoxication/withdrawal delirium. Organic psychoses patients had the longest lengths of stay and significantly more admissions to nonmedical/surgical units and discharges to nursing homes; almost 20% were African American. The recorded rate of delirium in the VA health system likely underestimates true prevalence and possibly reflects nonrecognition of delirium in many older veterans. Certain motoric and etiologic types of delirium may be more commonly diagnosed and recorded. Future research should prospectively examine recognition of motoric and etiologic delirium subtypes and racial differences in delirium diagnoses. PMID- 12641372 TI - Long-term effects of donepezil on P300 auditory event-related potentials in patients with Alzheimer's disease. AB - The P300, one of the cognitive event-related potentials (ERPs) of the cerebral cortex, reflects the functioning of the neurochemical system involved in cognitive processes. We investigated clinical significance of the components of auditory P300 ERPs, in comparison with neuropsychologic tests including the Mini Mental State Examination and the Japanese version of the Alzheimer's Disease Assessment Scale-cognitive subscale (ADAS-J cog), for evaluating of the effect of donepezil (DPZ) (5 mg daily for 6 months), an acetylcholinesterase inhibitor, in patients with Alzheimer's disease (AD). Reduction of P300 latency associated with a parallel improvement of ADAS-J cog scores was observed after administration of 5 mg/day of DPZ in patients with AD. P300 latency gives very useful information on the progression of AD, especially in the longitudinal follow-up of patients with AD during treatment with DPZ acting on cholinergic pathways. PMID- 12641373 TI - Age-related impairments in comprehending affective prosody with comparison to brain-damaged subjects. AB - This study evaluated the ability to produce and comprehend affective prosody across age groups and compared patterns of impaired performance to deficits observed after focal brain damage. Sixty-nine healthy subjects, ages 22 to 83 years, were given the Aprosodia Battery, a test that distinguishes between affective prosodic processing deficits following right- versus left-brain damage through the use of stimuli with progressively reduced verbal articulatory content. Production of affective prosody, measured by variation in fundamental frequency, was unimpaired in older subjects, whereas comprehension of affective prosody was impaired, particularly for tasks with reduced verbal articulatory content. The pattern of performance across affective comprehension tasks in the older subjects resembled the pattern found after right-brain damage. The results demonstrate age-related loss in comprehension of affective prosody that is most likely due to a processing deficit involving the right hemisphere. PMID- 12641374 TI - Effects of cognitive challenge on gait variability in patients with Parkinson's disease. AB - Attention plays an important role in the gait disturbances of patients with Parkinson's disease (PD); however, the effects of "dual tasking" on fall risk and gait instability have not been well studied. The authors tested the hypothesis that gait variability increases when subjects with PD walk while performing a cognitively challenging task (CCT). Subjects with idiopathic PD walked under normal conditions and while performing a CCT. During normal walking, gait variability was significantly (.0001 < P < .05) correlated with fall risk, disease duration, disease severity, motor function, mentation, behavior and mood, and cognitive function. When walking while cognitively challenged, gait variability increased (from 47 +/- 29 msec to 223 +/- 281 msec, P < .002). During dual-task walking, only disease duration remained significantly associated with gait variability. These results highlight the profound effects of attention on gait and indicate that walking while cognitively challenged impairs the ability of patients with PD to maintain a stable walk. PMID- 12641376 TI - Childhood abuse history, secondary traumatic stress, and child welfare workers. AB - Social workers are exposed to trauma vicariously through the trauma of their clients. This phenomenon, called secondary traumatic stress, vicarious traumatization, or compassion fatigue, presents a risk of negative personal psychological consequences. Based on a sample of 166 child welfare workers and using standardized measures, the study findings document the link between a personal history of primary trauma, childhood abuse or neglect, and the heightened risk for secondary traumatic stress in child welfare workers. PMID- 12641377 TI - Predictors of short-term reunification in South Australian substitute care. AB - This study examines the factors that contribute to short-term reunification for 235 children placed in South Australian substitute care from 1998 to 1999. Proportional hazard analyses showed that non-Aboriginal children and those placed because of parental incapacity were significantly more likely to go home, whereas neglected and Aboriginal children were significantly less likely to go home. A detailed profile analysis highlighted the specific case characteristics or changes thought to have contributed to reunification in each case. PMID- 12641378 TI - Illinois's child welfare research agenda: an approach to building consensus for practice-based research. AB - This article presents a consensus-building initiative to develop a statewide research agenda that responds to the needs of Illinois's child welfare community. Researchers conducted this process through a university-community partnership to engage those interested in child welfare services. The process and findings resulted in a living document that will guide child welfare research throughout the state. Findings of the inquiry suggest that the development of best practice models may be one of the most important contributions research can make to practice. PMID- 12641375 TI - Familial dementia with Lewy bodies with an atypical clinical presentation. AB - The authors report a case of a 64-year-old male with Alzheimer's disease (AD) and dementia with Lewy bodies (DLB) pathology at autopsy who did not manifest the core symptoms of DLB until very late in his clinical course. His initial presentation of early executive and language dysfunction suggested a cortical dementia similar to frontotemporal lobar degeneration (FTLD). Core symptoms of DLB including dementia, hallucination, and parkinsonian symptoms were not apparent until late in the course of his illness. Autopsy revealed both brainstem and cortical Lewy bodies and AD pathology. Family history revealed 7 relatives with a history of dementia including 4 with possible or probable DLB. This case is unique because of the FTLD-like presentation, positive family history of dementia, and autopsy confirmation of DLB. PMID- 12641379 TI - A first look at the need for enhanced support services for kinship caregivers. AB - This article describes findings from a series of focus groups conducted with formal kinship caregivers in Maryland. The findings reveal that kinship caregivers are committed to providing safety and stability for the children placed with them. The findings also suggest that caregiving can be a significant adjustment for many kinship caregivers and that expanded support services are needed to enhance their relationship with the child welfare agency. PMID- 12641380 TI - The Clara cells activated by acetaminophen. AB - Many high mountaineers die of acute mountain sickness in the high mountains of Nepal during their adventure. Most of them administer acetaminophen (AP) as an analgesic. Various cells with higher expression of cytochrome P-450 (CYP) would metabolize AP and lead to cellular impairment. CYP is a major enzyme system in the metabolism of xenobiotics in different organs of the human being including in Clara cells of the lungs. Clara cells contain the highest concentration of CYP. We injected AP to C57BL/6J mice to examine the toxicity of AP in Clara cells. In 8 hours Clara cells show more prominent apical protrusion with edematous smooth endoplasmic reticulum (sER), reduced secretary granules, and edema. We speculate that using AP in a high mountain area might be harmful for Clara cells being a trigger of high altitude sickness. PMID- 12641382 TI - Vibration analysis of obturator prostheses with different bulb height designs. AB - The purpose of this study was to investigate the effect on the vibratory characteristics of a cast hollow obturator prosthesis retainer when varying its bulb height. Bulb parts with high (H), middle (M), and low (L) lateral walls were prepared. A Vibration Generator was used to excite the three obturators, while a Laser-Doppler Vibrometer was employed to detect the vibrations at specified measurement points. The frequency response functions of each obturator were recorded on an FFT analyzer to enable their modal shapes to be identified. In addition, transient response simulations were carried out and the decay rate (DR) and the maximum amplitude (MA) of the rest and clasp parts of each obturator were obtained. These were statistically analyzed by ANOVA and Fisher's PLSD test (p < 0.01). The modal shapes were almost the same in all obturators. Significant differences were found in DR in all pairs among the three obturator prostheses, with the DR of the L type being the highest. On the other hand, there were no significant differences in MA. We conclude that the L type is preferable from the standpoint of its vibratory characteristics. PMID- 12641381 TI - High extracellular calcium affects osteoclastogenesis in mouse bone marrow cell culture. AB - The effects of high extracellular calcium (high Ca) in the local microenvironment on osteoclasts, osteoclast progenitors and stromal cells are not fully understood. We examined high Ca effect on osteoclastogenesis in mouse bone marrow cell culture. Mouse bone marrow cells were cultured for up to 6 days in the medium supplemented with 1, 25(OH)2 vitamin D3 (D3). High Ca treatment at the early stage of culture (the initial 24 hours) reduced the number of tartrate resistant acid phosphatase-positive multinuclear cells (TRAP(+)MNCs). This treatment slightly up-regulated the mRNA expressions of receptor activator of NF (B ligand (RANKL), RANK and osteoprotegerin (OPG). This inhibitory effect on the formation of TRAP(+)MNCs was recovered by RANKL. In contrast, high Ca treatment at the later stage of osteoclastogenesis (the last 2 days of culture) stimulated the formation of TRAP(+)MNCs, increased RANKL and RANK mRNA expressions and decreased OPG mRNA. High Ca at neither the early nor the later stage of culture affected the total number of adherent cells and the mRNA expression of alkaline phosphatase and osteopontin. In conclusion, high Ca affects osteoclastogenesis in a manner depending on the stage of osteoclastogenesis, which is partly mediated via the RANKL-RANK-OPG regulatory system. PMID- 12641383 TI - Study on the effects of shortening the distal end of a mouthguard using modal analysis. AB - Using a modal analysis system, we carried out a comparative assessment of the vibratory properties of the maxillary dentition of a human dry skull with a different distal end of a mouthguard. Mouthguards used in this study were: 1. a mouthguard covering up to the third molar (8MG), 2. the second molar (7MG), 3. the first molar (6MG), and 4. the second premolar (5MG). To identify the modal shape, the frequency response functions were recorded on a fast Fourier transform analyzer from the force signal impacted with a vibration generator and the response signal picked up using a laser-doppler vibrometer. The transient response waves were obtained throughout the transient response simulation procedures after curve-fitting procedures. Mean decay rates (sec(-1)) of the transient response waves were calculated for the anteriors, premolars and molars, and compared among different sets of experimental conditions. The decay rates became lower as the distal end of a mouthguard was short. The decay rates with 8MG, 7MG and 6MG (anteriors) exceeded the no mouthguard's decay rate + 2S.D. values. These results imply that a mouthguard should distally cover up to the second molar at least to ensure efficient absorption and/or dispersion of a traumatic force. PMID- 12641384 TI - Mechanism underlying the prevention of aneurismal rupture by coil embolization. AB - OBJECT: Endovascular treatment with Guglielmi detachable coil has been developed as a less invasive treatment for cerebral aneurysm. The aim of this study is to clarify the mechanism of the preventive effect of coil embolization. METHOD: Two aneurysm models were employed. One was a T-shaped bifurcation tube with a spherical dome made of glass, which was used for the measurement of pressure and visualization of flow pattern. The other model was a T-shaped glass tube with a spherical elastic silicone dome, which was used for the measurement of aneurismal wall displacement due to pulsation of flow. RESULT: 1, Guglielmi detachable coil caused no change in intra-aneurismal fluid pressure. 2, Coil insertion obstructed and slowed intra-aneurismal flow. This flow stagnation in the aneurysm might promote thrombus formation. 3, With increase in numbers of coils anchored at the intra-aneurismal wall, the displacement of the wall was considerably depressed. This suggests that coil insertion decreases the stress on the aneurismal wall. CONCLUSION: 1, Coil insertion depresses the pulsatile aneurysm wall movement, and diminishes the stress of the aneurysm wall. 2, Coil insertion obstructs intra aneurismal flow and facilitates thrombus formation in the aneurysm. These two factors may operate synergistically to prevent aneurysm rupture. PMID- 12641385 TI - Influence of patients perception on their acceptance and understanding of dental care within an education system. AB - This study examined the effects of conditions (that is, the fee structure and the easy of obtaining a dental appointment) on patient understanding and cooperation in clinical practice for dental hygiene and dental students. A questionnaire was given sequentially to 1483 patients attending the Tokyo Medical and Dental University dental hospital on "clinical education and patient satisfaction". Direct participants consisted of 650 patients, of which 213 (32.8%) were male and 422 (64.9%) female. The remaining 15 (2.3%) did not specify their gender. Patients who were satisfied with care received by dentists tended to be older compared to patients who were dissatisfied with dentists. The difference was significant (p < 0.001). Acceptable conditions for patients such as fees and appointments at the clinical session had an effected on patient acceptance of clinical training. The findings of this study suggest that patient satisfaction should be considered as part of the reasons for patients accepting dental care by students in their clinical education programs. Fees and appointment schedules strongly affect patient acceptance of clinical education. PMID- 12641386 TI - Effects of a denture adhesive on masticatory functions for complete denture wearers--consideration for the condition of denture-bearing tissues--. AB - The purpose of this study was to examine effects of a denture adhesive on masticatory functions for complete denture wearers considering the condition of denture-bearing tissues. Sixteen edentulous subjects wearing well-fitting complete dentures volunteered to participate in this study. According to the condition of denture-bearing tissues, subjects were divided into two groups; "good group" and "poor group". Maximum biting forces, masticatory performance, and electromyography of the masseter muscle during mastication were recorded with and without a denture adhesive. Durations of chewing burst and cycle, and coefficients of variation for these variables were calculated using electromyography recordings. Data were analyzed by using two-way repeated measured ANOVA and paired t-test in order to assess the effect of the use of a denture adhesive. The use of the denture adhesive increased maximum biting force and provided rhythmic masseter muscle activity during mastication for both groups. Masticatory performance was improved and duration of chewing burst was decreased only for "poor group". It was concluded that the effects of the denture adhesive on masticatory functions were observed overall for both groups, and more significant for denture wearers with poor denture-bearing tissues than with good denture-bearing tissues. PMID- 12641387 TI - Respiratory muscle stretch gymnastics in patients with post coronary artery bypass grafting pain: impact on respiratory muscle function, activity, mood and exercise capacity. AB - A new rehabilitation (New-RH) program including respiratory muscle stretch gymnastics (RMSG) was developed to alleviate post-coronary artery bypass grafting pain (PCP). Effects on respiratory muscle function, pain, activities of daily living (ADL), mood and exercise capacity were investigated. Subjects were 16 consecutive patients undergoing median full sternotomy coronary artery bypass grafting (CABG), and were randomly divided into equal New-RH (S-group) and conventional therapy (C-group) groups. Rib cage dominant breathing was observed postoperatively in both groups. With preoperative tan deltaVrc/deltaVab, increases at 1-week postoperatively and decreases at discharge for S-group tended to exceed those of C-group (p > .05). Decreased maximum inspiratory and expiratory pressure status for functional residual capacity and percent forced expiratory volume in one second at discharge again only tended to be smaller for S-group (p > .05). S-group displayed significantly reduced pain around both scapulas at discharge (p = .049), and increased mean overall ADL and profile of mood states (POMS)/Vigor scores (p = .031 and p = .018, respectively). POMS/Tension-Anxiety scores at discharge for S-group were significantly smaller than those preoperatively (p = .025), and S-group displayed significantly increased distance walked over 6-minutes at discharge than C-group (p = .029). New-RH improves patient participation in exercise therapy and increases exercise capacity by reducing PCP, relieving anxiety and tension, and improving ADL. PMID- 12641388 TI - Complete denture treatments for a cerebral palsy patient by using a treatment denture. A case report. AB - This article describes the prosthodontic treatment for a patient with cerebral palsy, in which complete dentures were successfully stabilized using treatment dentures. A 69-year-old edentulous male with no medical complications or mental retardation presented to our clinic. Opening movement of the jaw was possible, but a conspicuous mandibular shift towards the right was observed. He had never received any prosthodontic treatment. Initially, treatment dentures with flat tables were fabricated to rectify his erratic mandibular movements. During the first 3 weeks, the treatment dentures functioned poorly. Eventually, the patient could make tapping movement to some degree and have a meal with less effort. Indentation marks from the cusps of the opposing maxillary denture could be clearly seen on the flat tables. After six weeks, as he did not complain of any pain, definitive dentures were fabricated. When flat table treatment dentures are used, it is considered that the mucosa provides information regarding the vertical stop and bite force. In addition, it is speculated that there is an increase in the response from masseter muscle. In the present case, flat tables were effective for rehabilitation of the mandibular movement. PMID- 12641389 TI - Click-evoked ABR at high stimulus repetition rates for neonatal hearing screening. AB - A new, fast screening algorithm based on auditory brainstem response (ABR) recorded at a high click repetition rate is proposed. Response detection is carried out in the frequency domain by a statistical test procedure which includes the fundamental frequency and the harmonics below 800 Hz. First, the method was tested in 25 young adults. ABRs were recorded in the repetition rate range 20/s to 400/s. With a mean response detection time of 31 s, a click repetition rate of 140/s was found to be the optimum rate among the adult group. The method was then tested using a group of 114 neonates in whom the repetition rate range 60/s to 200/s was examined. At the repetition rate 90/s, which was found to be the optimum rate in neonates, the mean detection time was 24.6 s. In addition to the fast ABR detection, the proposed screening algorithm also allows simultaneous hearing screening of both ears using a one-channel data recording. PMID- 12641390 TI - Stacked tone-burst-evoked auditory brainstem response (ABR): preliminary findings. AB - In 1997, Don and his co-workers described a new method for evaluating the auditory brainstem response (ABR)--yielding the stacked-wave-V ABR--which may permit the reliable detection of even small vestibular schwannomas (VSs). However, this method requires a masking technique that may not be readily available to the clinician. Furthermore, relatively high-level noise is required and may be annoying to the patient. An alternative method that might permit the same manipulation of the ABR was thus explored whereby tone-bursts (TBs) were used to obtain frequency-specific ABRs. The goal of the study was to determine whether TBs could be used to achieve the sort of de-convolution and subsequent enhancement of wave V reported with the original method of Don et al. First, the TB-stacked ABR (wave V) was compared to that obtained using derived narrowband masked ABRs in young normal-hearing subjects. Second, the TB method was employed in subjects with unilateral VSs and in patients with unilateral sensory hearing loss in a preliminary clinical trial. The observed results are promising for the use of the TB method as an alternative approach to ABR 'stacking' and warrant further research and development of the TB method. PMID- 12641391 TI - Neuro-otological findings in Pendred syndrome. AB - Pendred syndrome is an autosomal recessive inherited disorder characterized by profound hearing impairment and inappropriate iodine release by the thyroid on perchlorate challenge. Thirty-three cases comprising members of 13 families and eight isolated cases were studied, with detailed audiological and vestibular investigation and computerized tomography. A uniform, profound, symmetrical sensorineural hearing loss was identified in all cases. Approximately one-third of the group reported progressive hearing impairment, in childhood or adolescence, associated with head injury, infection, or delayed secondary hydrops. Ninety per cent of the cases scanned showed dilated vestibular aqueducts, and all cases with progression of the hearing impairment demonstrated this structural abnormality. Approximately one-third of the cases had normal vestibular function, but a further third demonstrated a unilateral peripheral deficit, while the remaining third showed bilateral vestibular hypofunction. There was no intra-familial concordance of vestibular findings, and no correlation between vestibular abnormality and presence or absence of a dilated vestibular aqueduct, with or without a Mondini malformation. In older children and adults, Pendred syndrome was associated with a profound, symmetrical, sensorineural auditory impairment, and a variety of vestibular abnormalities, which are not uniform within families, or correlated with structural labyrinthine deformities. PMID- 12641392 TI - Hearing loss induced by noise, ear infections, and head injuries: results from the Nord-Trondelag Hearing Loss Study. AB - Pure-tone audiometry was administered to the adult population in Nord-Trondelag County, Norway, 1995-97. The 51,975 participants also provided questionnaire information about occupational and leisure noise exposure, previous ear infections, and head injury. Values averaged over both ears were calculated for low (250 and 500 Hz), middle (1 and 2 kHz) and high frequencies (3, 4, 6 and 8 kHz). The aim was to estimate the magnitude of hearing loss associated with various types of exposure by age and sex. Noise, ear infections and head injury explained 1-6% of the variance in hearing loss (varying with age, sex, and frequency range), in addition to what could be explained by age alone (30-58%). Only moderate effects of noise could be detected among women. The upper tenth percentile regarding occupational noise among men older than 44 years had on average an 8-9-dB high-frequency loss, adjusted for other predictors. Exposure to impulse noise (hunting, sports shooting) caused a 7-8-dB high-frequency loss in the same group. No significant effects of frequent use of personal stereo players or regular attendance at discotheques or rock concerts could be demonstrated. There were clear effects of recurrent ear infections and head injury. PMID- 12641393 TI - Hearing confirms existence and identity--experiences from persons with presbyacusis. AB - The purpose of the present qualitative study was to describe how elderly persons with presbyacusis experience living with that type of hearing loss. The ultimate goal is for these experiences to be used in personal-adjustment counselling in audiological rehabilitation. The study included seven men and seven women with mild-to-moderate hearing impairment of the typical presbyacusis type. Open-ended interviews were conducted with each person. The interviewees were analysed according to the phenomenographic approach, and 10 categories emerged: 'Conversation takes away or maintains identity', 'It's other people's fault that I can't hear', 'Other people make you realize you can't hear', 'Society makes you think you shouldn't mind about your hearing loss', 'It's natural to hear badly when you are old', 'You should hear well all your life', 'You want to keep a feeling of continuity in your daily life in spite of your hearing loss', 'You don't need to hear everything', 'You want to hear so you feel that you're alive', and 'You want to hear so you understand and keep yourself informed'. All these categories deal with identity or existence and form the basis for how the hearing impairment is experienced and managed. The subjects protected their identity in various ways, but above all by blaming their poor hearing on old age, and managing it with simple everyday strategies that did not break the feeling of continuity in everyday life. Not until they experienced the lack of sound as a lack of contact with life was there any interest in help in the form of hearing technology. There is a need for information about the possibility of rehabilitating presbyacusis, as hearing is important not only for communication and spatial orientation, but also as affirmation of our existence as human beings. PMID- 12641394 TI - Estrogen receptor action. AB - Today we have to consider the existence of two estrogen receptors, alpha and beta, in our attempts to understand the role of estrogen receptors in physiology and pathology, and to explain the pharmacology of estrogens, antiestrogens, and SERMs. Both ERalpha and ERbeta belong to the large family of nuclear receptors that function as hormone-dependent transcription factors with important roles in the endocrine signaling system. Estrogen signaling is dependent on multiple mechanisms involving the two estrogen receptors and, perhaps, also other, non estrogen receptor-related proteins. In the post-genomic era, with the help of technological advancements, we will see a dramatic increase in our understanding of the biological role of the two estrogen receptors, and in the function of estrogens, antiestrogens, and SERMs, which will significantly enhance our chances to develop designer drugs with dramatically improved therapeutic profiles. PMID- 12641395 TI - Development of gene-based therapies for cartilage repair. AB - Articular cartilage is particularly vulnerable to injury and degenerative conditions, and has a limited capacity for self-repair. Although current clinical procedures cannot restore a normal articular surface, there are a growing number of proteins that may be used to augment a repair process, or protect cartilage from degeneration. Because proteins are often difficult to administer effectively, gene therapy approaches are being developed to provide their sustained synthesis at sites of injury or disease. To promote cartilage repair, cDNAs can be targeted to synovium, or cartilage. Gene transfer to the synovium is generally considered more suitable for chondroprotective therapies that rely on expression of large amounts of anti-inflammatory mediators. The delivery of genes to cartilage defects to promote enhanced repair can be performed by either direct administration of gene delivery vectors, or by implantation of genetically modified chondrogenic cells. Variations of these methods have been used to demonstrate that exogenous cDNAs encoding growth factors can be delivered locally to sites of cartilage damage where they are expressed at physiologically relevant levels. Data is beginning to emerge that suggests that delivery and expression of these genescan influence a repair response toward the synthesis of normal articular cartilage in vivo. This article reviews the current status of gene delivery for cartilage healing and presents some of the remaining challenges. PMID- 12641396 TI - Genetic analysis of pain mechanisms. AB - Pain is the major reason for visits to the doctor. It is a complex sensation, with both a sensory and an emotional component. Its detailed regulation at all levels of the central nervous system, from the periphery to the cortex, has hindered our understanding of the neurobiological basis of pain. Furthermore, injury can produce long-lasting changes in pain pathways and chronic pain. The recent use of genetically modified organisms has significantly advanced our comprehension of the molecular and cellular mechanisms underlying pain. In this article we present the current state of knowledge regarding pain transmission, modulation, and plasticity, and some of the contributions made by studies of genetically altered mice. The cellular mechanisms explaining observed phenotypes, the involvement of supraspinal areas, and the plasticity underlying chronic pain are the three areas just beginning to be explored. PMID- 12641397 TI - Functional genomics in rat models of hypertension: using differential expression and congenic strains to identify and evaluate candidate genes. AB - Hypertension is a leading contributor to cardiovascular diseases such as heart attack and stroke. Genetic and environmental factors contribute to the development of hypertension. Animal models have been developed to study the genetic contributions to blood pressure (BP) regulation and to identify chromosomal regions harboring candidate genes causative of differences in BP regulation (i.e., BP quantitative trait loci [QTL]). Advances in both mammalian genome projects and global gene expression analysis present opportunities to study functional genomics in these animal models. In this article, novel approaches for designing experiments and interpreting global gene expression data using the Dahl salt-sensitive hypertension rat model are presented. We describe two-step screening protocols that can be used to identify BP QTL candidate genes. Genetically determined expression differences are identified in the target organs of inbred strains of contrasting phenotype in the first screen. Expression patterns in a panel of congenic strains or expression differences stemming from gene x environment interactions are examined in the second screen. Chromosomal locations of these genes can then be examined to determine whether they map to BP QTL-containing regions. Another approach is to study the expression of genes identified from public databases to be located within BP QTL-containing congenic regions. Several candidate genes have been identified using these strategies. PMID- 12641398 TI - A contrarian view of malaria therapy policy in Africa. PMID- 12641399 TI - Comparative efficacy of aminoquinoline-antifolate combinations for the treatment of uncomplicated falciparum malaria in Kampala, Uganda. AB - Resistance to chloroquine (CQ) requires its replacement as first-line therapy for uncomplicated malaria in much of Africa. Combination therapy may improve efficacy and delay the selection of resistant malaria parasites. Combinations of sulfadoxine-pyrimethamine (SP) with 4-aminoquinolines offer affordable and available alternatives to CQ. We conducted a randomized, single-blinded trial to compare the efficacy of SP monotherapy with combinations of SP and either CQ or amodiaquine (AQ) for the treatment of uncomplicated falciparum malaria in patients over 6 months of age in Kampala, Uganda. Of the 448 patients enrolled, 428 (95%) completed follow-up. Clinical treatment failure after 14 days occurred in 21/140 (15.0%, 95% CI 9.5-22.0%) SP-treated, 11/152 (7.2%, 95% CI 3.7-12.6%) SP/CQ-treated, and 0/136 (0%, 95% CI 0-2.7%) SP/AQ-treated patients. Combination therapies were safe and offered superior efficacy to SP monotherapy. SP/AQ was the most efficacious. This low-cost combination regimen may provide an optimal alternative to CQ for the treatment of uncomplicated malaria in Uganda. PMID- 12641400 TI - Distinguishing recrudescence from reinfection in a longitudinal antimalarial drug efficacy study: comparison of results based on genotyping of msp-1, msp-2, and glurp. AB - Genotyping frequently is used to distinguish recrudescent from new infections in antimalarial drug efficacy trials, but methodology and interpretation of results have not been standardized. We compared the utility of polymorphisms within 3 Plasmodium falciparum genes during a longitudinal trial in Kampala, Uganda. Merozoite surface protein-1 (msp-1) and merozoite surface protein-2 (msp-2) revealed greater diversity than glutamate-rich protein. Genotypes based on msp-1, msp-2, and all 3 genes combined were compared for 394 initial and subsequent isolates. Classification of most episodes as due to recrudescence or reinfection was straightforward. In 24% (msp-1), 16% (msp-2), and 62% (3 genes combined) of samples, subsequent episodes contained identical and new alleles, however. Our analysis suggested that such episodes should be classified as reinfections and not recrudescence. Comparing the 3 studied genes, msp-2 results were most accurate, and analysis of this single gene effectively distinguished recrudescence from reinfection in our study population. PMID- 12641401 TI - Drug-resistant malaria in Bangladesh: an in vitro assessment. AB - Forty-four Plasmodium falciparum isolates from Bangladesh and 22 from western Thailand were successfully tested for their drug susceptibility. High degrees of resistance were observed against chloroquine with geometric mean IC50s of 114.25 and 120.5 nM, respectively, for Bangladesh and western Thailand. Most isolates from both sites were sensitive to quinine, and all were sensitive to artesunate. Many isolates were considered in vitro resistant to mefloquine, but the geometric mean IC50 for the Thai isolates (98.79 nM) was 1.6 times (P = 0.002) higher than that of isolates from Bangladesh (60.3 nM). The high prevalence of in vitro mefloquine resistance in Bangladesh suggests that close surveillance is necessary to delay widespread multidrug resistant problems in the area. PMID- 12641402 TI - Association of Plasmodium falciparum chloroquine resistance transporter variant T76 with age-related plasma chloroquine levels. AB - The mutation leading to the substitution of a threonine (T76) for a lysine at position 76 of the Plasmodium falciparum chloroquine resistance transporter (PfCRT) was genotyped in 100 Nigerian children with asymptomatic parasitemia. Isolates containing both pfcrt variants were found to harbor higher numbers of parasite clones (P < 0.002). The prevalence of the pfcrt T76 variant decreased with age (P < 0.0001) and increased with blood levels of chloroquine (CQ) (P < 0.0001). Whereas the K76 allele was more frequent in individuals without detectable plasma CQ levels (79.7%), only the pfcrt T76 variant was observed in children with CQ levels > 150 nmol/L. In individuals without detectable plasma CQ, the proportion of those with pfcrt T76 decreased from 60% in children < 2 years old to 23.5% in children > or = 6 years old (P < 0.01). The association of actual blood levels of CQ and the occurrence of pfcrt T76 underlines that the pfcrt T76 variant is in fact the mediator of CQ resistance. PMID- 12641403 TI - Recrudescence in artesunate-treated patients with falciparum malaria is dependent on parasite burden not on parasite factors. AB - Artemisinin derivatives are first-line antimalarial drugs in Thailand. No firm evidence of clinically relevant artemisinin resistance exists. When used as monotherapy, artesunate has been associated with a high treatment failure (recrudescence) rate, which could be due to low-level artemisinin resistance. To understand the causes of recrudescence, we retrospectively studied a cohort of 104 malaria patients treated with artesunate monotherapy, 32 of whom recrudesced. There was no difference in in vitro artesunate sensitivities between 6 nonrecrudescent isolates and 16 paired admission and recrudescent isolates. Paired admission and recrudescent isolates from 10 patients were genotyped; only 3 had pfmdr1 mutations. Patients with admission parasitemias >10,000 per microl had a 9-fold higher likelihood of recrudescence (adjusted odds ratio) compared with patients with lower parasitemias. This study suggests (1) recrudescence after treatment with artesunate is not the result of inherent parasite resistance, and (2) admission parasitemia may be useful in choosing therapeutic options. PMID- 12641404 TI - Descriptive study on the efficacy and safety of artesunate suppository in combination with other antimalarials in the treatment of severe malaria in Sudan. AB - Documentation on the efficacy of artesunate in Africa is limited, and no experience of artesunate use in Sudan is documented. Severe malaria in rural areas of Sudan, where facilities for the safe and effective use of parenteral quinine are lacking, is a frequent problem. Early treatment with artesunate suppositories would provide a simple method for use by unskilled staff and would be an alternative approach to treat malaria in settings with poor resources. We describe a hospital-based study of rectal artesunate in 100 adult patients with severe falciparum malaria with a dose derived from pharmacokinetic data (200 mg every 8 hours) over 3 days, which halted progression of severe disease and had a low fatality rate. The dosage schedule led to a rapid clinical response and reduced parasite clearance and fever subsidence times of (31.5 +/- 10.1 hours) and (31.4 +/- 11.1 hours). The sequential treatment of rectal artesunate with either doxycycline or pyrimethamine/sulfadoxine or mefloquine resulted in similar clinical cure rates of around 100%, and the combination of artesunate with either doxycycline or pyrimethamine/sulfadoxine was equally effective as mefloquine in preventing recrudescence. There were no significant adverse effects or signs of toxicity related to the treatment observed during the 28-day follow-up. The combination regimens could be used in areas where there is limited access to parenteral therapy for malaria. PMID- 12641405 TI - Case report: An unusual late relapse of Plasmodium vivax malaria. AB - We observed an unusual case of Plasmodium vivax malaria who presented with an initial relapse four years after the primary infection. This occurred in Cameroon, where the patient, a 56-year-old priest, acquired a mild form of malaria and was treated with only chloroquine. Since he returned to Italy, he had not experienced any malaria-like symptoms, had not visited any other areas endemic for malaria, and had not received a blood transfusion. Blood smear microscopy confirmed the presence of Plasmodium spp. parasites, but unclear morphologic characteristics did not allow discrimination between P. vivax and P. ovale. A nested polymerase chain reaction-based molecular analysis identified P. vivax as the plasmodial species responsible. This case emphasizes the importance of taking into account the possibility of a very late initial relapse of P. vivax malaria and the relevant issues in terms of infection control. PMID- 12641406 TI - Transmission of mixed Plasmodium species and Plasmodium falciparum genotypes. AB - We studied malaria transmission by comparing parasite populations in humans and mosquito vectors at the household level. Blood samples were collected from all inhabitants for microscopic detection of gametocytes and polymerase chain reaction analysis. The next morning, blood-fed resting mosquitoes were collected inside the bed nets used by the individuals surveyed the previous afternoon. After 8 days of maintenance, mosquitoes were dissected, and midguts and salivary glands were recovered for polymerase chain reaction analysis. Results showed that parasite distribution was the same in the 2 hosts when compared at each household but was different when whole populations were analyzed. Different associations of Plasmodium species seem to occur in humans (Plasmodium falciparum/Plasmodium malariae) and mosquitoes (P. falciparum/Plasmodium ovale). Regarding P. falciparum infections, a higher proportion of single-genotype infections and less allele diversity are observed in mosquitoes than in humans. PMID- 12641407 TI - Malaria transmission in urban sub-Saharan Africa. AB - The rapid increase in the world's urban population has major implications for the epidemiology of malaria. A review of malaria transmission in sub-Saharan African cities shows the strong likelihood of transmission occurring within these sprawling cities, whatever the size or characteristics of their bioecologic environment. A meta-analysis of results from studies of malaria transmission in sub-Saharan Africa shows a loose linear negative relationship between mean annual entomologic inoculation rates (EIR) and the level of urbanicity. Few studies have failed to find entomologic evidence of some transmission. Our results show mean annual EIRs of 7.1 in the city centers, 45.8 in periurban areas, and 167.7 in rural areas. The impact of urbanization in reducing transmission is more marked in areas where the mean rainfall is low and seasonal. Considerable variation in the level of transmission exists among cities and within different districts in the same city. This article presents evidence from past literature to build a conceptual framework to begin to explain this heterogeneity. The potential for malaria epidemics owing to decreasing levels of natural immunity may be offset by negative impacts of urbanization on the larval ecology of anopheline mosquitoes. Malaria control in urban environments may be simpler as a result of urbanization; however, much of what we know about malaria transmission in rural environments might not hold in the urban context. PMID- 12641408 TI - Strong association between house characteristics and malaria vectors in Sri Lanka. AB - The objective of this study was to determine whether house characteristics could be used to further refine the residual insecticide-spraying program in Sri Lanka. Indoor-resting mosquito densities were estimated in 473 houses based on fortnightly collections over a two-and-a-half-year period. The type of house construction and the exact location of all houses were determined. In a multivariate analysis, distance of less than 750 meters between a house and the main vector-breeding site was strongly associated with the presence of Anopheles culicifacies in the house (odds ratio [OR] 4.8, 95% confidence interval [CI] 3.4 6.8) and to a lesser extent with the presence of An. subpictus (OR 1.4, 95% CI 1.1-1.7). Poor housing construction also was an independent risk factor (OR for An. culicifacies 1.3, 95% CI 1.0-1.9; OR for An. subpictus 1.3, 95% CI 1.0-1.6). It is recommended that a malaria control strategy focus on residential areas within 750 meters of streams and rivers, with special attention given to areas with the poorest type of house construction. PMID- 12641409 TI - Normal riboflavin status in malaria patients in Gabon. AB - Previous publications reported commonly the occurrence of riboflavin deficiency and a positive correlation between riboflavin status and parasitemia in patients with Plasmodium falciparum malaria. In these studies, riboflavin status was determined by erythrocyte glutathione reductase activation coefficients (EGRACs). Inherited low erythrocyte glutathione reductase activity is highly prevalent in malarial regions, however. To rule out falsely diagnosed riboflavin deficiency in affected patients, we conducted an investigation using a high-performance liquid chromatography method (HPLC) instead of the EGRAC method. In 29 infants (age range, 1-5 years), 22 schoolchildren (age range, 6-12 years), and 33 adolescents and adults (age range, 13-74 years) from Lambarene, Gabon, with acute P. falciparum malaria, plasma concentrations of riboflavin, flavin mononucleotide (FMN), and flavin adenine dinucleotide (FAD) were measured by HPLC. Results were correlated with parasite densities. Profiles of plasma concentrations of all 3 flavin compounds were within the normal range in all patients. Concentrations of free riboflavin were not different between the 3 age groups. In adolescents and adults, FMN and FAD concentrations were higher than in infants (P = 0.002 and P = 0.001) and schoolchildren (P = 0.003 and P = 0.002). Comparing children with hyperparasitemic and uncomplicated malaria, no difference in the concentrations of either flavin compound was found. Neither the concentrations of free riboflavin nor the concentrations of one of the flavin nucleotides correlated with parasitemia within subgroups of age or of children with uncomplicated and hyperparasitemic malaria. Our data indicate that nutritional riboflavin deficiency might have been overestimated in previous malaria studies and do not support a relationship between flavin concentrations and parasitemia in P. falciparum malaria. PMID- 12641411 TI - Serotype-specific dengue virus circulation and dengue disease in Bangkok, Thailand from 1973 to 1999. AB - Dengue virus circulation and association with epidemics and severe dengue disease were studied in hospitalized children with suspected dengue at the Queen Sirikit National Institute of Child Health in Bangkok, Thailand, from 1973 to 1999. Dengue serology was performed on all patients and viral isolation attempted on laboratory-confirmed patients. Acute dengue was diagnosed in 15,569 children and virus isolated from 4,846. DEN-3 was the most frequent serotype in primary dengue (49% of all isolates), DEN-2 in secondary and in dengue hemorrhagic fever (37% and 35%, respectively). The predominant dengue serotype varied by year: DEN-1 from 1990-92, DEN-2 from 1973-86 and 1988-89; DEN-3 in 1987 and 1995-99; and DEN 4 from 1993-94. Only DEN-3 was associated with severe outbreak years. Our findings illustrate the uniqueness of each serotype in producing epidemics and severe disease and underscore the importance of long-term surveillance of dengue serotypes in understanding the epidemiology of these viruses. PMID- 12641410 TI - Human genetic polymorphisms and asymptomatic Plasmodium falciparum malaria in Gabonese schoolchildren. AB - Several studies have focused their attention on the relationship between host genetic factors and susceptibility/resistance to severe malaria. However, there is a paucity of information concerning the role of host genetic factors in asymptomatic malaria, a form of low-grade Plasmodium falciparum infection without clinical symptoms. We investigated in this study the potential relationship between the host (human) genetic polymorphisms (glucose-6-phosphate dehydrogenase [G6PD], mannose binding lectin [MBL], tumor necrosis factor alpha [TNFalpha]( 308) and (-238), and nitric oxide synthase 2 [NOS2](-954)) and the prevalence and profile of asymptomatic P. falciparum infection in 158 Gabonese schoolchildren. We found that G6PD A- heterozygous females (18 of 74) have a low prevalence of asymptomatic malaria (38.9% versus 67.3%; P = 0.03, by chi-square test). Children heterozygous for TNFalpha(-238) (25 of 156) carry high number of diverse infecting parasite genotypes (2.5 versus 1.99; variance F = 3.05). No statistically significant association was found between MBL, TNFalpha(-308), or NOS2 polymorphisms and asymptomatic malaria. Upon combining our data on asymptomatic forms with those from the literature for others forms, we conclude that G6PD A- heterozygous females are protected against all forms of P. falciparum malaria, and that the TNFalpha(-238A) allele confers protection against clinical malaria. PMID- 12641412 TI - Low oral receptivity for dengue type 2 viruses of Aedes albopictus from Southeast Asia compared with that of Aedes aegypti. AB - Dengue hemorrhagic fever has been a major health problem in Asia since the 1950s. During this period, the former principal vector of dengue viruses in Asia, Aedes albopictus, was replaced by Aedes aegypti in most major cities of the area. Ae. aegypti is now considered the main vector of dengue viruses in Asia. Surprisingly, however, this mosquito has been described as having a relatively low oral receptivity for dengue viruses compared with Ae. albopictus. In the present study, we compared the relative oral receptivities of Ae. aegypti and Ae. albopictus collected in southeast Asia from both sympatric and allopatric breeding sites. In all instances, the oral receptivity of Ae. aegypti to the dengue type 2 virus used was significantly higher than that of Ae. albopictus. We also compared the relative oral receptivity of Ae. aegypti and Ae. albopictus for two other low-passage strains of dengue 2. In all instances, Ae. aegypti was significantly more receptive than Ae. albopictus. It should be noted, however, that the difference was found only for Ae. albopictus recently collected from the field (Ta Promh strain, Cambodia, 2001) and not for an Ae. albopictus strain that had been colonized for many years (Oahu strain, Hawaii, 1971). We also observed a significant increase in the infection rate of Ae. albopictus of the Ta Promh strain with increasing generations in the laboratory. These observations demonstrate the importance of considering the colonization history of mosquitoes when assessing their susceptibility to infection with dengue viruses and, perhaps, other arboviruses. PMID- 12641413 TI - Dengue and its vectors in Thailand: calculated transmission risk from total pupal counts of Aedes aegypti and association of wing-length measurements with aspects of the larval habitat. AB - Working in a village dengue focus in Chachoengsao Province, Thailand, aedine mosquito larvae and pupae were counted in all containers of 10 houses per month. The wings of female Aedes aegypti (L.) emerging from pupae were measured. Number of pupae and size of emerging females increased in containers with qualities that favored availability of larval food sources (e.g., uncovered containers). The small size of most mosquitoes compared with those raised in the laboratory indicated that the larval population as a whole was under nutritional stress. Applying the number of pupae per house and measurement of air and water temperature with an existing model, the risk of dengue transmission was greatest in May and June. The estimated number of female Ae. aegypti per house was well above the threshold for increasing transmission in all months but December through February. A phased approach to sampling immature aedine mosquitoes in Thailand is proposed, which would consist of routine surveillance of larval index and occasional total counts with measurement of wing size. Such a system would combine the benefits of the simple application of larval surveillance with the valuable data gathered from pupal counts and wing measurements. PMID- 12641414 TI - Limited potential for mosquito transmission of genetically engineered, live attenuated western equine encephalitis virus vaccine candidates. AB - Specific mutations associated with attenuation of Venezuelan equine encephalitis (VEE) virus in rodent models were identified during efforts to develop an improved VEE vaccine. Analogous mutations were produced in full-length cDNA clones of the Cba 87 strain of western equine encephalitis (WEE) virus by site directed mutagenesis in an attempt to develop an improved WEE vaccine. Isogenic viral strains with these mutations were recovered after transfection of baby hamster kidney cells with infectious RNA. We evaluated two of these strains (WE2102 and WE2130) for their ability to replicate in and be transmitted by Culex tarsalis, the principal natural vector of WEE virus in the United States. Each of the vaccine candidates contained a deletion of the PE2 furin cleavage site and a secondary mutation in the E1 or E2 glycoprotein. Both of these potential candidates replicated in mosquitoes significantly less efficiently than did either wild-type WEE (Cba 87) virus or the parental clone (WE2000). Likewise, after intrathoracic inoculation, mosquitoes transmitted the vaccine candidate strains significantly less efficiently than they transmitted either the wild-type or the parental clone. One-day-old chickens vaccinated with either of the two vaccine candidates did not become viremic when challenged with virulent WEE virus two weeks later. Mutations that result in less efficient replication in or transmission by mosquitoes should enhance vaccine safety and reduce the possibility of accidental introduction of the vaccine strain to unintentional hosts. PMID- 12641415 TI - Entomological evaluation by pool screen polymerase chain reaction of Onchocerca volvulus transmission in Ecuador following mass Mectizan distribution. AB - The prevalence of infected and infective black flies was estimated by pool screen polymerase chain reaction (PCR) amplification in the three river basins in Ecuador endemic for onchocerciasis. Mass distribution of ivermectin (Mectizan) resulted in dramatic declines in the prevalence of infected and infective flies. In the Rio Santiago river basin, no infections were detected, suggesting that transmission had ceased. The ratio of infected to infective flies in Simulium exiguum was 10-fold lower than the corresponding ratio for Simulium quadrivittatum, suggesting that S. exiguum is a more-competent vector for Onchocerca volvulus than S. quadrivittatum. However, the prevalence of infective flies in the two species was not different, suggesting that S. quadrivittatum may play an important vectorial role where it is the dominant human-biting species. The data demonstrate that pool screen PCR is an efficient way to monitor transmission in areas subject to control, and to certify an area as free of O. volvulus transmission. PMID- 12641416 TI - Detection of Cryptosporidium parvum and Giardia lamblia carried by synanthropic flies by combined fluorescent in situ hybridization and a monoclonal antibody. AB - Wild-caught synanthropic flies were tested for the presence of Cryptosporidium parvum and Giardia lamblia on their exoskeletons and in their digestive tracks by fluorescent in situ hybridization and fluorescein isothiocyanate (FITC) conjugated monoclonal antibody (MAb) against Cryptosporidium and Giardia cell wall epitopes. The levels of C. parvum were positively correlated with the levels of G. lamblia, indicating a common source of contamination. The majority of oocysts and cysts were potentially viable (C. parvum = 80% and G. lamblia = 69%). More G. lamblia cysts occurred on the exoskeleton of the flies than within the digestive tracts; the opposite relationship was observed for C. parvum. No genotype other than C. parvum G2 was found to be associated with flies. Because filth flies carry viable C. parvum oocysts and G. lamblia cysts acquired naturally from unhygienic sources, they can be involved in the epidemiology of cryptosporidiosis and giardiasis. Fluorescent oligonucleotide probes used together with FITC-conjugated MAb represent a convenient and cost-effective technique for rapid and specific identification of human-infectious species of Cryptosporidium and Giardia mechanically transported by flies, and for the assessment of the viability of these pathogens. PMID- 12641417 TI - Short report: Treatment failure in Hansen's disease. AB - Areas of low endemicity of Hansen's disease, such as Texas, California, and Hawaii, exist due to immigration and rare autochthonous infections. Managing this disease in these areas of low endemicity is difficult, especially in observing for relapse. The accurate diagnosis of relapse is imperative so that appropriate therapy can be promptly reinstituted and unnecessary treatment can be avoided. To assess treatment failures in an area of low endemicity, we retrospectively evaluated 113 patients with Hansen's disease treated in southern Texas. Of 57 patients who completed therapy, 11 were later restarted on medications for this disease for presumed relapse. However, nine of the 11 were found not to have true relapses of Hansen's disease. The accurate diagnosis of relapse of this disease is essential not only in the individual patient but also for prospective treatment trials to establish best practices. PMID- 12641418 TI - Association and epidemiologic features of Trypanosoma cruzi and human T cell lymphotropic a virus type II in inhabitants of the Paraguayan Gran Chaco. AB - Serologic evidence of Trypanosoma cruzi infection was demonstrated in 43.5% of 519 Paleoamerindians and in only 2.5% of 161 non-Indians (Mennonites of German descent and Paraguayans of Spanish descent) inhabiting an area of western Paraguay that belongs to the Gran Chaco territory. These people ranged in age between two and 80 years. All were also tested for infection with the human T cell lymphotropic virus type II (HTLV-II). The prevalence of HTLV-II infection was 22.1% in Indians and 3.7% in non-Indians. As determined by a multivariate logistic regression analysis that controlled for relevant confounders, an HTLV-II infected individual was 2.28 times more likely to be seropositive for T. cruzi than an HTLV-II negative. Possible explanations for this finding are discussed. The difference in T. cruzi prevalence between Indians and non-Indians was associated with differences between these groups in exposure to known risk factors for infection with the parasite. There were significant differences in the seroprevalence of T. cruzi among the two predominant Indian groups, even when they inhabited communities that were close to each other. These differences were associated with differences in the prevalence of HTLV-II infection but not with differences in exposure to known risk factors for T. cruzi infection. Infection with T. cruzi increased with age, was greater in males than in females, and clustered in families. PMID- 12641419 TI - Analysis of the presence of Trypanosoma cruzi in the heart tissue of three patients with chronic Chagas' heart disease. AB - It is still unclear to what extent myocarditis-associated, chronic Chagas' heart disease is due to persisting Trypanosoma cruzi. In the present study, we have analyzed tissue samples from the hearts of three patients with this disease. In situ hybridization provided little evidence for the presence of intact T. cruzi even at sites of strong inflammation. Nevertheless, micromanipulation techniques detected remnants of both T. cruzi kinetoplast DNA and nuclear DNA. Trypanosoma cruzi DNA was also detected in single macrophages dissected directly from frozen heart tissue sections. Thus, this analysis demonstrates that T. cruzi kinetoplast DNA and nuclear DNA are widely dispersed in the heart tissue, although in low amounts. Since we rarely detected intact T. cruzi parasites during the chronic phase of Chagas' heart disease, we can exclude heart tissue as a major parasite reservoir. PMID- 12641420 TI - Morphologic and functional characterization of Chagasic heart disease in non human primates. AB - Chagasic heart disease has been documented in non-human primates, but noninvasive characterization of systolic and diastolic function has not been previously reported. Seventeen seropositive (12 females; mean age, 20) and 13 age- and gender-matched seronegative baboons underwent Doppler echocardiography. Systolic function indices included left ventricular (LV) fractional shortening (FS %), velocity of circumferential fiber shortening (VCF, circ/sec), LV mass index, and left and right ventricular ejection fractions (RVEF %). Diastolic function indices included transmitral E-wave, A-wave, E/A ratio, E-deceleration time, and isovolumic relaxation time. Twelve-lead electrocardiographic (ECG) recordings were obtained. There were no significant differences between groups for body size or blood pressure. Seropositive and seronegative groups revealed diffuse non specific T wave changes precluding differentiation; however, tall "P" waves were seen in four seropositive and two seronegative baboons. Four of the 17 (24%) seropositive baboons had decreased FS (25 +/- 8% versus 40 +/- 5%, P < 0.005) and VCF (1.05 +/- 0.36 circ/sec versus 1.84 +/- 0.23 circ/sec, P < 0.0001), prolonged isovolumic relaxation time (71 +/- 16 msec versus 55 +/- 9 msec, P < 0.02), and reduced RVEF (44 +/- 9% versus 54 +/- 4%, P < 0.05), as compared with the other seropositive baboons. We conclude that chagasic heart disease is present in 24% of the naturally infected baboons in this study. ECG evidence of right atrial enlargement was more common in the seropositive animals. There were systolic and diastolic abnormalities of both ventricles. The LV systolic dysfunction may be segmental or diffuse. PMID- 12641422 TI - The association of health-care use and hepatitis C virus infection in a random sample of urban slum community residents in southern India. AB - To determine whether health-care use was associated with prevalent hepatitis C virus (HCV) infection in Chennai, India, 1,947 adults from 30 slum communities were randomly selected to be interviewed about parenteral and sexual risks for HCV infection and to provide biological specimens for HCV and sexually transmitted infection (STI) testing. Prevalent HCV infection was detected in 2.4% of non-injection drug using (IDU) participants. Controlling for other associated factors, and excluding IDU, men who used informal health-care providers were five times as likely to be HCV infected as those who did not use informal providers (Adjusted Odds Ratio, AOR = 5.83; 95% confidence interval [CI]: 1.57, 21.6), a finding not detected in women. More research is needed to determine the extent to which HCV infection is associated with reuse of contaminated injection equipment in health-care settings in developing countries. PMID- 12641421 TI - Multiple hemorrhagic brain infarcts after viper envenomation. AB - We report the case of a 65-year-old woman who presented with severe neurologic complications after envenomation by a viper snake. A computed tomography (CE) scan revealed multiple brain hemorrhagic infarcts. Conservative treatment in this case proved to be sufficient and repetitive CT scans displayed a complete resolution of the radiologic findings. Possible mechanisms for the cerebral infarctions are discussed. The mechanism of infarctions in this case was believed to be the vasomotor and coagulation disorders caused by the toxins present in the snake's venom and was one of the reasons that led to conservative treatment. PMID- 12641423 TI - Ciguatera poisoning in Vanuatu. AB - Ciguatera poisoning is endemic in many tropical and subtropical countries. We conducted a retrospective study of admissions to two hospitals on the islands of Vanuatu in the southwestern Pacific region. We estimated the annual hospital admission rate for fish poisoning to be 65 (95% confidence interval [CI] = 55 75)/100,000 population on the island of Santo and 29 (95% CI = 19-43)/100,000 population on the island of Ambae. Hospital admission was more common in males 20 29 years old. Death was a rare complication. In the face of increases in both tourism and in the global trade in tropical and exotic fish, physicians in both endemic and non-endemic areas should be familiar with the epidemiology and clinical features of this important condition. PMID- 12641424 TI - Origins of individual variability in P4501A induction. PMID- 12641425 TI - Selenium compounds modulate the activity of recombinant rat AsIII methyltransferase and the methylation of arsenite by rat and human hepatocytes. AB - Formation of methylated metabolites is a critical step in the metabolism of inorganic arsenic or selenium. We have previously shown that under conditions of a concurrent exposure sodium selenite inhibits methylation of arsenite by cultured rat hepatocytes. Here, we compare the effects of sodium selenite and mono-, di-, and trimethylated selenium compounds on the methylation of arsenite by purified recombinant rat As(III)-methyltransferase (Cyt19) and by primary rat and human hepatocytes. Among these compounds, sodium selenite was the most potent inhibitor of the methylation of arsenite by the recombinant enzyme (K(i) = 1.4 microM) and by cultured cells. In both systems, methylseleninic acid was an order of magnitude less potent an inhibitor (K(i) = 19.4 microM) than was sodium selenite. Dimethylselenoxide and trimethylselenonium iodide were weak activators of recombinant As(III)-methyltransferase activity but were weak inhibitors of arsenite methylation in hepatocytes. These data suggest that selenite, rather than its methylated metabolites, is responsible for inhibition of arsenite methylation in cultured hepatocytes and that inhibition may involve direct interactions between selenite and As(III)-methyltransferase. PMID- 12641426 TI - Adduction of the chloroform metabolite phosgene to lysine residues of human histone H2B. AB - Human exposure to trihalomethanes such as chloroform has been associated with both cancer and reproductive toxicity. While there is little evidence for chloroform mutagenicity or DNA adduct formation, in vivo studies in rats have demonstrated adduction to histones and other nuclear proteins. Histones play a key role in controlling DNA expression particularly through the acetylation of lysine residues in their N-termini. Therefore, we studied the reaction of phosgene, the major active metabolite of chloroform, with the N-terminus of human histone H2B (Hpep, Pro-Glu-Pro-Ala-Lys-Ser-Ala-Pro-Ala-Pro-Lys-Lys-Gly-Ser-Lys Lys-Ala-Val-Thr-Lys-Ala-Gln-Lys) in a model chemical system. The aim of this study was to assess whether phosgene is able to form irreversible adducts with this peptide and to investigate which residues are most susceptible. Hpep was reacted with a range of phosgene concentrations (0.03-36 mM) at 37 degrees C, pH 7.4. The products of these reactions, analyzed by matrix-assisted laser desorption ionization MS, showed that up to three CO moieties could be adducted to the peptide. The singly and doubly adducted peptides were purified by HPLC and then hydrolyzed with trypsin to produce a series of fragments that were analyzed by HPLC-MS. The tryptic products showed that adduction occurred principally at lysine residues, and that all seven lysine residues of the peptide were subject to adduction. Collision-induced dissociation analysis using ion trap MS-MS of the tryptic fragment [Pro-Glu-Pro-Ala-Lys-Ser-Ala-Pro-Ala-Pro-Lys + CO] and of the full-length singly adducted peptide supported the role of lysine residues in adduction; the data also indicated that the N-terminal proline and the serine residues are susceptible. Addition of glutathione to the reaction mixture only partially attenuated adduct formation and allowed production of another adducted species, i.e., Hpep-CO-glutathione. The occurrence of such reactions to the N termini of histones, if confirmed by in vivo studies, could help to explain the mechanism of chloroform carcinogenicity. PMID- 12641427 TI - Synthesis, cytotoxicity, and QSAR analysis of X-thiophenols in rapidly dividing cells. AB - In this study, the cytotoxicities of a series of X-thiophenols vs rapidly growing mouse leukemia cells in vitro are determined. The resulting ID(50) values are then used to formulate a quantitative structure-activity relationship, which is well-correlated by the Brown variation of the Hammett electronic parameter, sigma plus (sigma(+) ), such that Log 1/ID(50) = -0.93 (+/-0.18) sigma(+) + 0.86 (+/ 0.24) I(H) + 3.99 (+/-0.13). I(H) represents an indicator variable that calls attention to the unusual activity of halogens and pseudohalogens. In lieu of sigma(+), homolytic bond dissociation energies (BDE) are also used successfully to correlate the cellular cytotoxicities of thiophenols. The nature of substituent effects on cellular toxicity is examined, and they reveal that electron-releasing substituted thiophenols such as 4-amino thiophenol and the 4 alkoxy thiophenols are highly cytotoxic and effective at inhibiting cellular proliferation at physiological pH. On the other hand, electron-attracting substituted thiophenols such as the 4-cyano and 4-halogen analogues show a reduced ability to inhibit the cell growth of this cell line. Thus, there is a clear parallel between enhanced biological activity and electron releasing ability as measured by sigma(+) constants or BDE values. The susceptibility of the cellular interaction to electronic effects as delineated by the coefficient with the sigma(+) term (also called the Hammett rho value) is high (-0.96), suggesting that substantial energetic assistance is provided by the substituents and that a weak initiating radical reactant such as superoxide radical may be involved. Previous cytotoxicity studies of a large diverse data set of X-phenols in this cell line and embryo cells have also revealed a more pronounced dependence on sigma(+) and deltaBDE. A comparison of reaction constants obtained from thiophenoxy radical formation reactions and phenoxy radical formation reactions in organic media suggests radical-mediated involvement in cell cytotoxicity. Such cells could be more vulnerable to the effects of reactive thiyl species on their metabolism and subsequent proliferation. PMID- 12641428 TI - Synthesis and characterization of N-demethylated metabolites of malachite green and leucomalachite green. AB - Malachite green (MG), a triphenylmethane dye used to treat fungal and protozoan infections in fish, undergoes sequential oxidation to produce various N demethylated derivatives (monodes-, dides(sym)-, dides(unsym)-, trides-, and tetrades-) both before and after reduction to leucomalachite green (LMG). The close structure resemblance of the metabolites with aromatic amine carcinogens implicates a potential genotoxicity from exposure to MG. The availability of the synthetic standards is important for metabolic and DNA adduct studies of MG. This paper describes a simple and versatile method for the synthesis of MG, LMG, and their N-demethylated metabolites. The synthesis involves a coupling of 4 (dimethylamino)benzophenone or 4-nitrobenzophenone with the aryllithium reagents derived from appropriately substituted 4-bromoaniline derivatives, followed by treatment with HCl in methanol. The resulting cationic MG and their leuco analogues showed systematic UV/vis spectral and tandem mass fragmentation patterns consistent with sequential N-demethylation. The extensive (1)H and (13)C spectral assignments of the metabolites were aided by the availability of (13)C(7)-labeled MG and LMG. The results indicate the existence of a resonance structure with the cationic charge located in the central methane carbon (C(7)). The synthetic procedure is general in scope so that it can be extended to the preparation of N-demethylated metabolites of other structurally related N methylated triphenylmethane dyes. PMID- 12641429 TI - An integrated metabonomic investigation of acetaminophen toxicity in the mouse using NMR spectroscopy. AB - An integrated metabonomics study using high-resolution 1H NMR spectroscopy has been applied to investigate the biochemical composition of intact liver tissue (using magic angle spinning), liver tissue extracts, and blood plasma samples obtained from control and acetaminophen-treated mice. Principal components analysis was used to visualize similarities and differences in biochemical profiles. The time- and dose-dependent biochemical effects of acetaminophen were related to the drug toxicity, as determined using histopathology. Metabolic effects in intact liver tissue and lipid soluble liver tissue extracts from animals treated with the high dose level of acetaminophen included an increase in lipid triglycerides and monounsaturated fatty acids together with a decrease in polyunsaturated fatty acids, indicating mitochondrial malfunction with concomitant compensatory increase of peroxisomal activity. In addition, a depletion of phospholipids was observed in treated liver tissue, which suggested an inhibition of enzymes involved in phospholipid synthesis. There was also a depletion in the levels of liver glucose and glycogen. In addition, the aqueous soluble liver tissue extracts from high dose animals also revealed an increase in lactate, alanine, and other amino acids, together with a decrease in glucose. Plasma spectra showed increases in glucose, acetate, pyruvate, and lactate. These observations all provide evidence for an increased rate of glycolysis. These findings could indicate a mitochondrial inability to use pyruvate in the citric acid cycle and also reveal the impairment of fatty acid beta-oxidation in liver mitochondria of such treated mice. PMID- 12641430 TI - Spectroscopic characterization of the 4-hydroxy catechol estrogen quinones derived GSH and N-acetylated Cys conjugates. AB - Estrogens, including the natural hormones estrone (E(1)) and estradiol (E(2)), are thought to be involved in tumor induction. Specifically, catechol estrogen quinones (CEQs) derived from the catechol estrogens 4-hydroxyestrone (4-OHE(1)) and 4-hydroxyestradiol (4-OHE(2)) react with DNA and form DNA adducts (Cavalieri, E. L., et al. (1997) Proc. Natl Acad. Sci. U.S.A. 94, 10037). CEQs are also conjugated with GSH, a reaction that prevents damage to DNA, providing biomarkers of exposure to CEQs. Current detection limits for these analytes by HPLC with multichannel electrochemical detection are in the picomole range (Devanesan, P., et al. (2001) Carcinogenesis 22, 489). To improve the detection limit of CEQ derived conjugates, spectrophotometric monitoring was investigated. Fluorescence and/or phosphorescence spectra of the 4-OHE(1), 4-OHE(2), Cys, N-acetylcysteine (NAcCys), 4-OHE(1)-2-SG, and 4-OHE(2)-2-SG conjugates and their decomposition products 4-OHE(1)-2-NAcCys and 4-OHE(2)-2-NAcCys were obtained at 300 and 77 K. It is shown that (i) 4-OHE(1)- and 4-OHE(2)-derived SG and NAcCys conjugates are weakly fluorescent at 300 K (with the emission maximum at 332 nm) but strongly phosphorescent at 77 K; (ii) Cys and NAcCys exhibit fluorescence and phosphorescence only at 77 K; and (iii) 4-OHE(1) and 4-OHE(2) are weakly fluorescent at 300 and 77 K and not phosphorescent. The phosphorescence spectra of SG and NAcCys conjugates are characterized by a weak origin band at approximately 383 nm and two intense vibronic bands at 407 and 425 nm. After they are cooled from 300 to 77 K, the total luminescence intensity of SG and NAcCys conjugates increases by a factor of approximately 150 predominantly due to phosphorescence enhancement. Theoretical calculations revealed, in agreement with the experimental data, that the lowest singlet (S(1)) and triplet (T(1)) states of 4-OHE(2)-2-NAcCys are of n,pi* and pi,pi* character, respectively, leading to a large intersystem crossing yield and strong phosphorescence. The limit of detection (LOD) for CEQ-derived conjugates, based on phosphorescence measurements, is in the low femtomole range. The concentration LOD is approximately 10(-9) M. Therefore, we propose that capillary electrophoresis interfaced with low temperature phosphorescence detection can be used to test for human exposure to CEQs by analyzing urine. PMID- 12641432 TI - Identification of intermediate pathways of 4-hydroxynonenal metabolism in the rat. AB - The formation of 4-hydroxy-2-nonenal (HNE) conjugates with glutathione (GSH) by Michael addition and subsequent cleavage to yield the related mercapturic acid (MA) conjugates are a major detoxication process. To characterize the metabolic pathways involved in the formation of urinary HNE-MA conjugates in the rat, the metabolism of HNE-thioethers (HNE-GSH, HNE-MA, and HNE-Cys) by rat liver and kidney cytosolic fractions was investigated. The experimental results showed that HNE-GSH is a good substrate for cytosolic incubations whereas HNE-MA and HNE-Cys are poorly metabolized. About 80% of the urinary MA conjugates originate from the primary and major HNE metabolite, namely, the hemiacetalized HNE-GSH. The direct reduction of HNE-GSH by a cytosolic aldo-keto reductase (NADPH) leads to 1,4 dihydroxynonene-GSH (DHN-GSH) and subsequently to DHN-MA. The direct oxidation of HNE-GSH by aldehyde dehydrogenase (NAD)(+) leads to 4-hydroxynonenoic-lactone GSH, the partial hydrolysis of which occurs at physiological pH and accounts for the corresponding 4-hydroxynonenoic-GSH. Both the spontaneous- and the glutathione S-transferases-catalyzed retro-Michael cleavages of HNE-GSH and HNA lactone-GSH are the source of HNE and HNA-lactone, respectively. This latter compound, with both lipophilic and electrophilic properties, is available for microsomal omega-hydroxylation by cytochrome P450 4A enzymes and conjugation with thiol groups and therefore is the most likely candidate for the formation of omega-hydroxylated HNE-mercapturic acid conjugates excreted in rat urine. PMID- 12641431 TI - 11-Deoxy,16,16-dimethyl prostaglandin E2 induces specific proteins in association with its ability to protect against oxidative stress. AB - Prostaglandins (PGs) act locally to maintain cellular homeostasis and stimulate stress response signaling pathways. These cellular effects are diverse and are tissue-dependent. PGE(2), and the synthetic analogue, 11-deoxy,16,16-dimethyl PGE(2) (DDM-PGE(2)), protect renal proximal tubular epithelial (LLC-PK1) cells against cellular injury induced by the potent nephrotoxic and nephrocarcinogenic metabolite of hydroquinone, 2,3,5-tris-(glutathion-S-yl)hydroquinone. Although this cytoprotective response (in LLC-PK1 cells) is mediated through a thromboxane or thromboxane-like receptor coupled to AP-1 signaling pathways, the mechanism of cytoprotection is unknown. In this study, we utilized HPLC-electrospray ionization tandem mass spectrometric (ESI MS/MS) and matrix-assisted laser desorption ionization time-of-flight mass spectrometric (MALDI TOF) analysis of proteins isolated from DDM-PGE(2)-stimulated LLC-PK1 cells to identify candidate cytoprotective proteins. DDM-PGE(2) selectively stimulated the synthesis of several proteins in LLC-PK1 cells. Peptide sequencing by ESI-MS/MS of in-gel tryptic protein digests revealed the identity of eight proteins: endothelial actin binding protein, myosin, elongation factor 2 (EF-2), elongation factor 1alpha-1 (EF-1alpha), heat shock protein 90beta (HSP90beta), glucose-regulated protein 78 (GRP 78), membrane-organizing extension spike protein, and actin. Both ESI-MS/MS and MALDI-MS analysis resulted in the same protein identification. Western analysis confirmed the temporal induction of the majority of these proteins, including EF-2, EF-1alpha, HSP90beta, GRP78, and actin. The collective expression of these proteins suggests that DDM-PGE(2)-mediated cytoprotection may involve alterations in cytoskeletal organization and/or stimulation of an endoplasmic reticulum (ER) stress response. The present studies provide insights into potential downstream targets of PG signaling. PMID- 12641433 TI - Ascorbic acid modifies the surface of asbestos: possible implications in the molecular mechanisms of toxicity. AB - Ascorbic acid is one of the major components of the antioxidants defenses of the lung lining layer where inhaled asbestos fibers are deposited. Crocidolite fibers were incubated at 37 degrees C in a 0.01 M aqueous solution of ascorbic acid for 25 days in order to investigate modifications in surface reactivity. Iron (820 nmol/mg) and monomeric silica (470 nmol/mg) were released in the supernatant, while ascorbic acid was consumed. The amount of iron and silicon released, respectively, 17 and 6% (in atoms) of the total fiber content, exceeded what was expected at the surface, suggesting a partial disgregation of crocidolite promoted by ascorbic acid. In the absence of ascorbic acid but at the same pH, the release of iron and monomeric silica was minimal. At time intervals, aliquots of fibers were withdrawn to evidence chemical modifications progressively taking place. Three families of Fe(II) centers, differing in coordinative unsaturation and progressively removed during incubation, have been evidenced from the FTIR spectra of NO adsorbed onto the fibers. The most uncoordinated ones are removed first. New highly uncoordinated iron sites are exposed at the fiber surface as a consequence of the erosion of the outmost layers while hydration of silica tetrahedra yields new silanol groups. The activity in the Fenton-like reaction (*OH from H(2)O(2)) decreases following surface iron depauperation. Conversely, the homolytic cleavage of the C-H bond (CO(2)*-) from the formate ion) appears related to the small fraction of iron ions always present but easily quenched by the adsorption of ascorbic acid or its oxidation products. PMID- 12641434 TI - Metabolism of capsaicin by cytochrome P450 produces novel dehydrogenated metabolites and decreases cytotoxicity to lung and liver cells. AB - Capsaicin is a common dietary constituent and a popular homeopathic treatment for chronic pain. Exposure to capsaicin has been shown to cause various dose dependent acute physiological responses including the sensation of burning and pain, respiratory depression, and death. In this study, the P450-dependent metabolism of capsaicin by recombinant P450 enzymes and hepatic and lung microsomes from various species, including humans, was determined. A combination of LC/MS, LC/MS/MS, and LC/NMR was used to identify several metabolites of capsaicin that were generated by aromatic (M5 and M7) and alkyl hydroxylation (M2 and M3), O-demethylation (M6), N- (M9) and alkyl dehydrogenation (M1 and M4), and an additional ring oxygenation of M9 (M8). Dehydrogenation of capsaicin was a novel metabolic pathway and produced unique macrocyclic, diene, and imide metabolites. Metabolism of capsaicin by microsomes was inhibited by the nonselective P450 inhibitor 1-aminobenzotriazole (1-ABT). Metabolism was catalyzed by CYP1A1, 1A2, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, and 3A4. Addition of GSH (2 mM) to microsomal incubations stimulated the metabolism of capsaicin and trapped several reactive electrophilic intermediates as their GSH adducts. These results suggested that reactive intermediates, which inactivated certain P450 enzymes, were produced during catalytic turnover. Comparison of the rate and types of metabolites produced from capsaicin and its analogue, nonivamide, demonstrated similar pathways in the P450-dependent metabolism of these two capsaicinoids. However, production of the dehydrogenated (M4), macrocyclic (M1), and omega-1-hydroxylated (M3) metabolites was not observed for nonivamide. These differences may be reflective of the mechanism of formation of these metabolites of capsaicin. The role of metabolism in the cytotoxicity of capsaicin and nonivamide was also assessed in cultured lung and liver cells. Lung cells were markedly more sensitive to cytotoxicity by capsaicin and nonivamide. Cytotoxicity was enhanced 5 and 40% for both compounds by 1-ABT in BEAS-2B and HepG2, respectively. These data suggested that metabolism of capsaicinoids by P450 in cells represented a detoxification mechanism (in contrast to bioactivation). PMID- 12641435 TI - Major intermediates in organophosphate synthesis (PCl3, POCl3, PSCl3, and their diethyl esters) are anticholinesterase agents directly or on activation. AB - Three phosphotrichlorides [phosphorus trichloride (PCl(3)), phosphorus oxychloride (POCl(3)), and thiophosphoryl chloride (PSCl(3))] with an annual U.S. production of >500,000,000 pounds and their diethyl esters are intermediates in the production of organophosphorus pesticides, plastics, flame retardants, and hydraulic fluids. They are classified as highly toxic to mammals based on acute oral and inhalation data with rats. This study considers their mechanisms of toxicity. PCl(3) and POCl(3) inhibit acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) from several species with in vitro IC(50) values of 5-36 and 88-1200 microM, respectively; PSCl(3) is a less potent inhibitor. These phosphotrichlorides have high vapor toxicity to houseflies with in vivo inhibition of brain AChE activity correlating with mortality. PCl(3) and POCl(3) produce cholinergic poisoning signs on ip administration to mice, and all three phosphotrichlorides give marked in vivo inhibition of serum BChE but not brain AChE activity. PCl(3) is a direct acting AChE inhibitor. Our earlier proposed activation of POCl(3) is confirmed here by preparing pure Cl(2)P(O)OH and its potassium and dicyclohexylamine salts that reproduce the action of POCl(3) as in vitro AChE inhibitors and toxicants in mice. PSCl(3) on hydrolysis yields Cl(2)P(O)SH [which oxidizes with peracid to Cl(2)P(O)SOH] as the proposed activation product. Vapors of (EtO)(2)PCl, (EtO)(2)P(O)Cl, and (EtO)(2)P(S)Cl are lethal to houseflies as in vivo AChE inhibitors, the first two acting directly and the last one on oxidative activation to (EtO)(2)P(O)Cl (possibly by P450) or (EtO)(2)P(O)SCl (a phosphorylating agent in a peracid oxidation system). Thus PCl(3), (EtO)(2)PCl, and (EtO)(2)P(O)Cl act directly as AChE inhibitors whereas POCl(3) and PSCl(3) undergo hydrolytic activation and (EtO)(2)P(S)Cl undergoes oxidative activation. In contrast, the toxicity to mice of phosphofluorides [FP(O)Cl(2), F(Cl)P(O)OH, and F(2)P(O)OH; studied as model compounds for comparison] may be due to liberating fluoride ion. PMID- 12641436 TI - Quantification of tamoxifen DNA adducts using on-line sample preparation and HPLC electrospray ionization tandem mass spectrometry. AB - The nonsteroidal antiestrogen tamoxifen is used as an adjuvant chemotherapeutic agent for the treatment of all stages of hormone-dependent breast cancer and more recently as a chemopreventive agent in women with elevated risk of developing the disease. While clearly beneficial for the treatment of breast cancer, tamoxifen has been reported to increase the risk of endometrial cancer in women. Furthermore, it has been shown to be hepatocarcinogenic in rats. Tamoxifen is clearly genotoxic in rat liver, as indicated by the formation of DNA adducts; the occurrence of tamoxifen DNA adducts in human endometrial tissue is more controversial. The detection and quantitation of tamoxifen DNA adducts have relied primarily upon (32)P-postlabeling, with other techniques, such as immunoassays and accelerator mass spectrometry, being used to a much lesser extent. To expand the range of available analytical methodologies for quantifying tamoxifen DNA adducts, we have developed an assay using on-line sample preparation, coupled with HPLC and electrospray ionization tandem mass spectrometry (ES-MS/MS). alpha-Acetoxytamoxifen was reacted with salmon testis DNA at ratios between 0.1 ng and 1 mg alpha-acetoxytamoxifen per mg DNA. After enzymatic hydrolysis to nucleosides, the most highly modified DNA samples were analyzed by HPLC-UV, which indicated the presence of two adduct peaks in approximately a 1:4 ratio. The major adduct was isolated, rigorously characterized as (E)-alpha-(deoxyguanosin-N(2)-yl)tamoxifen, and quantified on the basis of its molar extinction coefficient. A similar reaction was conducted with [N(CD(3))(2)]-alpha-acetoxytamoxifen to prepare a deuterated adduct that could serve as an internal standard for ES-MS/MS. The limit of detection for the HPLC-ES-MS/MS method was approximately 5 adducts/10(9) nucleotides, with an intra and interassay precision of 3% relative standard deviation. The method was validated over the range of 8-1 520,000 adducts/10(8) nucleotides using 100 microg samples of DNA modified in vitro. Analysis of liver DNA from female Sprague-Dawley rats treated by gavage with seven daily doses of 20 mg tamoxifen/kg body weight gave a value of 496 +/- 16 adducts/10(8) nucleotides for (E)-alpha-(deoxyguanosin-N(2)-yl)tamoxifen and 626 +/- 18 adducts/10(8) nucleotides for (E)-alpha-(deoxyguanosin-N(2)-yl)-N-desmethyltamoxifen. These data indicate that the HPLC-ES-MS/MS methodology has sufficient sensitivity and precision to be useful in the analysis of tamoxifen DNA adducts formed in vivo in experimental models and may be able to detect tamoxifen DNA adduct formation in human tissue samples. PMID- 12641437 TI - Transformation and activation of benzidine by oxidants of the inflammatory response. AB - Aromatic amines, such as benzidine (BZ), initiate bladder cancer in humans. Inflammation/infection play an important role in this cancer. This study was designed to assess the influence of inflammatory oxidants, including reactive nitrogen oxygen species (RNOS), on BZ transformation and activation. RNOS were generated under various conditions and reacted with BZ, and the products were examined by HPLC. Conditions that generate nitrogen dioxide radical, NO(2)(-) + myeloperoxidase + H(2)O(2) and ONOO(-), produced primarily a single new product, which was identified by MS as azobenzidine (AZO-BZ). The myeloperoxidase catalyzed reaction was inhibited by 1 mM cyanide and did not require NO(2)(-). Chloride (100 mM) reduced the myeloperoxidase reaction by 30% with taurine having little effect. In contrast, conditions that generate N(2)O(3), i.e., NO donor diethylamine (DEA) NONOate, produced two products, which were identified by MS as 4'-OH-4-aminobiphenyl (4'-OH-ABP) and 4-aminobiphenyl (ABP). 2-(4-Carboxyphenyl) 4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, an oxidant of NO thought to produce NO(2)(*), had a biphasic effect on product formation. At a concentration equal to DEA NONOate, a 5-fold increase in BZ nitrosation was observed, while at higher concentrations nitrosation was greatly diminished and formation of AZO-BZ occurred. Glutathione prevented RNOS transformation of BZ. With MPO and ONOO(-), a new product was formed that cochromatographed with 3-(glutathione-S-yl)BZ. Glutathione also prevented nitrosation of BZ but did not form additional BZ products. HOCl-mediated activation of BZ, 4'-OH-ABP, and ABP to bind DNA was assessed. A higher level of binding was observed at pH 5.5 than pH 7.4. BZ elicited the most binding. More binding was observed at both pH values with 4'-OH ABP than ABP. Thus, components of the inflammatory response are capable of BZ transformation and activation. PMID- 12641438 TI - Studies on metabolic pathways of cocaine and its metabolites using microsome preparations from rat organs. AB - Cocaine metabolism has been studied previously with respect to the formation of predominant hydrolytic or hepatotoxic metabolites via oxidative pathways. In the present study, cocaine and eight of its metabolites (norcocaine, ecgonine methyl ester, benzoylecgonine, benzoylnorecgonine, 3-hydroxy-benzoylecgonine, cocaethylene, norcocaethylene, and ecgonine ethyl ester) were incubated with microsomes from rat liver, kidney, lung, and brain. Qualitative analysis of the metabolites produced was performed using solid phase extraction (SPE), trimethylsilylation, and GC/MS. It was found that the metabolites with a free carboxylic group (e.g., benzoylecgonine) were not further oxidized by microsomal enzymes and their presence in urine or blood may therefore be due to hydrolysis of the respective alkylated entities. Although microsomes from all organs exhibited oxidative metabolism, significant differences were noted. Kidney microsomes produced essentially the same results as liver, but aryl hydroxylated metabolites were not found in incubations with lung and brain microsomes. N Hydroxy-norcocaine was found only in traces with brain microsomes. It appears that cocaine is converted to N-hydroxy-norcocaine (which is the precursor of toxic metabolites) not only in the liver but also in other organs of rat. This might be relevant in the development of lung toxicity observed in smokers of cocaine ("crack"). PMID- 12641439 TI - Identification of products formed by reaction of 3',5'-di-O-acetyl-2' deoxyguanosine with hypochlorous acid or a myeloperoxidase-H2O2-Cl- system. AB - Hypochlorous acid (HOCl), generated by myeloperoxidase from H(2)O(2) and Cl(-), plays an important role in host defense and inflammatory tissue injury. We have studied the reaction of 3',5'-di-O-acetyl-2'-deoxyguanosine with reagent HOCl and with a human myeloperoxidase-H(2)O(2)-Cl(-) system in order to characterize polar reaction products. When 100 microM 3',5'-di-O-acetyl-2'-deoxyguanosine was reacted with 100 microM HOCl at pH 7.4 and 37 degrees C and the reaction was terminated by N-acetylcysteine, 3',5'-di-O-acetyl derivatives of previously reported products, such as diastereomers of spiroiminodihydantoin nucleoside, a diimino-imidazole nucleoside, an amino-imidazolone nucleoside, and 8-chloro-2' deoxyguanosine were formed. In addition, we report the formation of 3',5'-di-O acetyl derivatives of a guanidinohydantoin nucleoside, an iminoallantoin nucleoside, and a diamino-oxazolone nucleoside in this system. The identification of the products was based on their identical ESI-MS and UV spectra and HPLC retention times with authentic compounds synthesized with other oxidation systems. All of these products were also formed in the reaction of 3',5'-di-O acetyl-2'-deoxyguanosine with the myeloperoxidase-H(2)O(2)-Cl(-) system under mildly acidic conditions. The yields of the products were greatly affected by the pH of the reaction mixture. The total yields of these products formed by HOCl at pH 7.4 and by the myeloperoxidase-H(2)O(2)-Cl(-) system at pH 4.5 were 72 and 43% of the consumed 3',5'-di-O-acetyl-2'-deoxyguanosine, respectively, indicating that nearly half of the consumption of 3',5'-di-O-acetyl-2'-deoxyguanosine by HOCl and the myeloperoxidase-H(2)O(2)-Cl(-) system can be accounted for by the formation of these products. PMID- 12641440 TI - The mechanism of the peroxynitrite-mediated oxidation of myoglobin in the absence and presence of carbon dioxide. AB - Among the cellular components that can react directly with peroxynitrite in the presence of physiological CO(2) concentrations are sulfur-, selenium-, and metal containing proteins, in particular hemoproteins. We have previously shown that the reactions of peroxynitrite with oxymyoglobin (MbFeO(2)) and oxyhemoglobin proceed via the corresponding ferryl species, which, in a second step, are reduced to the iron(III) forms of the proteins (metMb and metHb). In this study, we have investigated the influence of the concentration of added CO(2) on the kinetics and the mechanism of the peroxynitrite-mediated oxidation of MbFeO(2). We found that this reaction proceeds in two steps via the formation of MbFe(IV)=O also in the presence of millimolar amounts of CO(2). As compared to the values measured in the absence of added CO(2), the second-order rate constant for the first reaction step in the presence of 1.2 mM CO(2) is significantly larger [(4.1 +/- 0.7) x 10(5) M(-1) s(-1), at pH 7.5 and 20 degrees C], whereas that for the peroxynitrite-mediated reduction of MbFe(IV)=O to metMb is almost unchanged [(3.2 +/- 0.2) x 10(4) M(-1) s(-1), at pH 7.5 and 20 degrees C]. Finally, we show that because of the parallel decay of peroxynitrite, 8-25 equiv are needed to completely oxidize MbFeO(2) to metMb, with larger amounts required to oxidize diluted MbFeO(2) solutions in the presence of CO(2). Simulation of the reaction in the absence and presence of CO(2) was carried out to get a better understanding of the mechanism. The results suggest that CO(3)*(-) and NO(2)* may be involved in the reaction and interact with MbFeO(2) and MbFe(IV)=O, respectively. PMID- 12641441 TI - Reactions of N,N-bis(2-chloroethyl)-p-aminophenylbutyric acid (chlorambucil) with 2'-deoxyadenosine. AB - N,N-bis(2-chloroethyl)-p-aminophenylbutyric acid (chlorambucil, 1; 0.6 mM) was allowed to react with 2'-deoxyadenosine (16.1 mM) at physiological pH (cacodylic acid, 50% base), and the reactions were followed by HPLC-MS and HPLC-MS/MS techniques. Although the predominant reaction observed was chlorambucil hydrolysis, ca. 7% of 1 reacted with various heteroatoms of the nucleoside. The principal site of alkylation was N1. Several other adducts were also detected. The N1, N6, N3, and N7 derivatives were characterized by means of MS/MS, UV, and (1)H NMR. The N6 adduct is derived directly from alkylation of N6 of 2'-dAdo. Dimroth rearrangement of the N1 adduct to the N6 adduct was very slow under the reaction conditions employed. Minor adducts such as a carbohydrate derivative were tentatively characterized by MS/MS. No cross-links were detected. The role of chlorambucil-2'-deoxyadenosine adducts in the cytotoxicity and mutagenicity of 1 is also discussed. PMID- 12641442 TI - Reactive oxygen species are involved in arsenic trioxide inhibition of pyruvate dehydrogenase activity. AB - Arsenite was shown to inhibit pyruvate dehydrogenase (PDH) activity through binding to vicinal dithiols in pure enzyme and tissue extract. However, no data are available on how arsenite inhibits PDH activity in human cells. The IC(50) values for arsenic trioxide (As(2)O(3)) to inhibit the PDH activity in porcine heart pure enzyme preparation and in human leukemia cell line HL60 cells were estimated to be 182 and 2 microM, respectively. Thus, As(2)O(3) inactivation of PDH activity was about 90 times more potent in HL60 cells than in purified enzyme preparation. The IC(50) values for As(2)O(3) and phenylarsine oxide to reduce the vicinal thiol content in HL60 cells were estimated to be 81.7 and 1.9 microM, respectively. Thus, As(2)O(3) is a potent PDH inhibitor but a weak vicinal thiol reacting agent in HL60 cells. Antioxidants but not dithiol compounds suppressed As(2)O(3) inhibition of PDH activity in HL60 cells. Conversely, dithiol compounds but not antioxidants suppressed the inhibition of PDH activity by phenylarsine oxide. As(2)O(3) increased H(2)O(2) level in HL60 cells, but this was not observed for phenylarsine oxide. Mitochondrial respiration inhibitors suppressed the As(2)O(3)-induced H(2)O(2) production and As(2)O(3) inhibition of PDH activity. Moreover, metal chelators ameliorated whereas Fenton metals aggravated As(2)O(3) inhibition of PDH activity. Treatment with H(2)O(2) plus Fenton metals also decreased the PDH activity in HL60 cells. Therefore, it seems that As(2)O(3) elevates H(2)O(2) production in mitochondria and this may produce hydroxyl through the Fenton reaction and result in oxidative damage to the protein of PDH. The present results suggest that arsenite may cause protein oxidation to inactivate an enzyme and this can occur at a much lower concentration than arsenite binding directly to the critical thiols. PMID- 12641444 TI - Inorganic and dimethylated arsenic species induce cellular p53. AB - Arsenic compounds are known for their ability both to cause and to treat human cancers, although the molecular mechanisms underlying these actions are incompletely understood. The simplest explanation is that arsenic causes DNA damage that leads to mutations. However, the majority of scientific evidence indicates that arsenic is not a genotoxin or DNA-damaging agent. DNA damage typically leads to cellular responses designed to minimize the replication of damaged DNA, such as the induction of p53, and p53 induction has therefore been used as an indicator of DNA damage. Because this approach can be applied to human cells and does not rely on a specific, heritable mutation occurring at a particular site, it seemed possible that this method could detect DNA damage that was undetectable using other techniques. To examine the genotoxic potential of arsenic compounds, therefore, seven of these compounds (sodium arsenite, sodium arsenate, methyloxoarsine, iododimethylarsine, disodium methyl arsonate, dimethylarsinic acid, and arsenic trioxide) were tested for their ability to increase the cellular level of p53 as measured by ELISA. Of this group, arsenic trioxide was the strongest inducer of cellular p53, while dimethylarsinic acid, iododimethylarsine, and sodium arsenite also caused p53 induction in a dose- and time-dependent manner. Sodium arsenate, as well as the two monomethyl compounds tested, methyloxoarsine and disodium methyl arsonate, did not cause detectable increases in cellular p53. Our results indicate, therefore, that cells respond to several of these arsenic compounds as they do to chemicals that damage DNA, suggesting that exposure of cells to these compounds does in fact cause DNA damage. Such damage could then result in mutations and the observed development of cancer. PMID- 12641443 TI - Naturally occurring coumarins inhibit human cytochromes P450 and block benzo[a]pyrene and 7,12-dimethylbenz[a]anthracene DNA adduct formation in MCF-7 cells. AB - Naturally occurring coumarins (NOCs) inhibit polycyclic aromatic hydrocarbon induced skin tumor initiation in mice by blocking cytochrome P450 (P450)-mediated bioactivation of benzo[a]pyrene (B[a]P) and 7,12-dimethylbenz[a]anthracene (DMBA). Bergamottin selectively inhibits tumor initiation by B[a]P, whereas imperatorin and isopimpinellin inhibit tumor initiation with both carcinogens. The goals of the current study were to examine the ability of NOCs to inhibit human P450s in vitro and to establish whether NOCs, which are anticarcinogenic in mice, can block carcinogen bioactivation in cultured human cells. For the initial experiments, incubations containing 5 microM P450, P450 substrate, an NADPH generating system, and NOCs were used to determine the concentrations of each inhibitor that blocked 50% of P450 activity (IC(50)). These results confirmed that NOCs are capable of inhibiting multiple human P450s and that they exhibit selectivity for certain isoforms of human P450s. In subsequent experiments, we examined the effects of bergamottin, imperatorin, and isopimpinellin on DMBA and B[a]P DNA adduct formation in the human breast MCF-7 adenocarcinoma cell line. Coincubation of cells with the three different NOCs significantly inhibited DMBA DNA adduct formation by 29-82% at doses ranging from 2 to 10 microM and significantly inhibited B[a]P DNA adduct formation by 37-80% at doses ranging from 20 to 80 microM. HPLC analysis of the DNA hydrolysates demonstrated that inhibition of DNA adducts corresponded to inhibition of the major B[a]P and DMBA diol-epoxide-derived adducts. Although bergamottin was not effective at blocking DMBA bioactivation in the mouse skin model, it was similar in effectiveness to imperatorin and isopimpinellin in MCF-7 cells. These results demonstrate that NOCs, which are present in citrus fruits and other components of the human diet, are capable of inhibiting carcinogen metabolizing enzymes and blocking bioactivation of both B[a]P and DMBA in MCF-7 cells. PMID- 12641446 TI - Insights into the amyloid folding problem from solid-state NMR. PMID- 12641447 TI - Dual-site recognition of different extracellular matrix components by anti angiogenic/neurotrophic serpin, PEDF. AB - Pigment epithelium-derived factor (PEDF), a member of the serine protease inhibitor (serpin) superfamily, possesses anti-angiogenic and neurotrophic activities. PEDF has been reported to bind to extracellular matrix (ECM) components such as collagens and glycosaminoglycans (GAGs). In this study, to determine the binding sites for collagens and GAGs, we analyzed the interaction of recombinant mouse PEDF (rPEDF) with collagen I and heparin. By utilizing residue-specific chemical modification and site-directed mutagenesis techniques, we revealed that the acidic amino acid residues on PEDF (Asp(255), Asp(257), and Asp(299)) are critical to collagen binding, and three clustered basic amino acid residues (Arg(145), Lys(146), and Arg(148)) are necessary for heparin binding. Mapping of these residues on the crystal structure of human PEDF (Simonovic, M., Gettins, P. G. W., and Volz, K. (2001) Proc. Natl. Acad. Sci. U.S.A. 98, 11131 11135) demonstrated that the collagen-binding site is oriented toward the opposite side of the highly basic surface where the heparin-binding site is localized. These results indicate that PEDF possesses dual binding sites for different ECM components, and this unique localization of ECM-binding sites implies that the binding to ECM components could regulate PEDF activities. PMID- 12641448 TI - Role of two residues proximal to the active site of Ubc9 in substrate recognition by the Ubc9.SUMO-1 thiolester complex. AB - The small ubiquitin-like modifier SUMO-1 is covalently attached to lysine residues on target proteins by a specific conjugation pathway involving the E1 enzyme SAE1/SAE2 and the E2 enzyme Ubc9. In an ATP-dependent manner, the C terminus of SUMO-1 forms consecutive thiolester bonds with cysteine residues in the SAE2 subunit and Ubc9, before the Ubc9.SUMO-1 thiolester complex catalyzes the formation of an isopeptide bond between SUMO-1 and the epsilon-amino group of the target lysine residue on the protein substrate. The SUMO-1 conjugation pathway bears many similarities with that of ubiquitin and other ubiquitin-like protein modifiers (Ubls), and because of its production of a singly conjugated substrate and the lack of absolute requirement in vitro for E3 enzymes, the SUMO 1/Ubc9 system is a good model for the analysis of protein conjugation pathways that share this basic chemistry. Here we describe methods of both steady-state and half-reaction kinetic analysis of Ubc9, and use these techniques to determine the role of two residues, Asp(100) and Lys(101) of Ubc9 which are not found in E2 enzymes from other protein conjugation pathways. These residues are found close to the active site Cys in the tertiary structure of Ubc9, and although they are shown to inhibit the transesterification reaction from SAE1/SAE2, they are important for substrate recognition in the context of the thiolester complex with SUMO-1. PMID- 12641449 TI - Structural analysis of the photosystem I supercomplex of cyanobacteria induced by iron deficiency. AB - Here we describe the three-dimensional structure of the newly discovered CP43' photosystem I (PSI) supercomplex of cyanobacteria calculated by single-particle analysis of images obtained by electron cryomicroscopy (cryo-EM). This large membrane protein complex has a molecular mass of approximately 2 MDa and is found in cyanobacteria when grown in iron deficient media. It is composed of a reaction center trimer surrounded by 18 subunits of the chlorophyll a binding CP43'protein, encoded by the isiA gene, which increases the light harvesting capacity of PSI by approximately 70%. By modeling higher-resolution structural data obtained from X-ray crystallography into the three-dimensional (3D) cryo-EM map, we have been able to gain a better understanding of the structure and functional properties of this supermolecular complex. We have identified three separate clusters of chlorophyll molecules at the periphery of the PSI core which may aid energy transfer from the CP43' antenna ring to the reaction center. Moreover, it is shown that despite the replacement of ferredoxin with flavodoxin as an electron acceptor under iron stress conditions, the 3D map has density to accommodate the extrinsic proteins, PsaC, PsaD, and PsaE. The presence of these three proteins was also confirmed by immunoblotting. PMID- 12641450 TI - Sterol carrier protein-2 functions in phosphatidylinositol transfer and signaling. AB - Over 20 years ago, it was reported that liver cytosol contains at least two distinct proteins that transfer phosphatidylinositol in vitro, phosphatidylinositol transfer protein (PITP) and a pH 5.1 supernatant fraction containing sterol carrier protein-2 (SCP-2). In contrast to PITP, there has been minimal progress on the structural and functional significance of SCP-2 in phosphatidylinositol transport. As shown herein, highly purified, recombinant SCP 2 stimulated up to 13-fold the rapid (s) transfer of radiolabeled phosphatidylinositol (PI) from microsomal donor membranes to highly curved acceptor membranes. SCP-2 bound to microsomes in vitro and overexpression of SCP 2 in transfected L-cells resulted in the following: (i) redistribution of phosphatidylinositols from intracellular membranes (mitochondria and microsomes) to the plasma membrane; (ii) enhancement of insulin-mediated inositol triphosphate production; and (iii) 5.5-fold down regulation of PITP. Like PITP, SCP-2 binds two ligands required for vesicle budding from the Golgi, PI, and fatty acyl CoA. Double immunolabeling confocal microscopy showed SCP-2 significantly colocalized with caveolin-1 in the cytoplasm (punctate) and plasma membrane of SCP-2 overexpressing hepatoma cells (72%), HT-29 cells (58%), and SCP 2 overexpressing L-cells (37%). Taken together, these data show for the first time that SCP-2 plays a hitherto unrecognized role in intracellular phosphatidylinositol transfer, distribution, and signaling. PMID- 12641451 TI - Specificity determinants of human cathepsin s revealed by crystal structures of complexes. AB - Cathepsin S, a lysosomal cysteine protease of the papain superfamily, has been implicated in the preparation of MHC class II alphabeta-heterodimers for antigen presentation to CD4+ T lymphocytes and is considered a potential target for autoimmune-disease therapy. Selective inhibition of this enzyme may be therapeutically useful for attenuating the hyperimmune responses in a number of disorders. We determined the three-dimensional crystal structures of human cathepsin S in complex with potent covalent inhibitors, the aldehyde inhibitor 4 morpholinecarbonyl-Phe-(S-benzyl)Cys-Psi(CH=O), and the vinyl sulfone irreversible inhibitor 4-morpholinecarbonyl-Leu-Hph-Psi(CH=CH-SO(2)-phenyl) at resolutions of 1.8 and 2.0 A, respectively. In the structure of the cathepsin S aldehyde complex, the tetrahedral thiohemiacetal adduct favors the S configuration, in which the oxygen atom interacts with the imidazole group of the active site His164 rather than with the oxyanion hole. The present structures provide a detailed map of noncovalent intermolecular interactions established in the substrate-binding subsites S3 to S1' of cathepsin S. In the S2 pocket, which is the binding affinity hot spot of cathepsin S, the Phe211 side chain can assume two stable conformations that accommodate either the P2-Leu or a bulkier P2-Phe side chain. This structural plasticity of the S2 pocket in cathepsin S explains the selective inhibition of cathepsin S over cathepsin K afforded by inhibitors with the P2-Phe side chain. Comparison with the structures of cathepsins K, V, and L allows delineation of local intermolecular contacts that are unique to cathepsin S. PMID- 12641452 TI - HIV-1 vaccine development: constrained peptide immunogens show improved binding to the anti-HIV-1 gp41 MAb. AB - The human immunodeficiency virus type I (HIV-1) transmembrane glycoprotein gp41 mediates viral entry through fusion of the target cellular and viral membranes. A segment of gp41 containing the sequence Glu-Leu-Asp-Lys-Trp-Ala has previously been identified as the epitope of the HIV-1 neutralizing human monoclonal antibody 2F5 (MAb 2F5). The 2F5 epitope is highly conserved among HIV-1 envelope glycoproteins. Antibodies directed at the 2F5 epitope have neutralizing effects on a broad range of laboratory-adapted HIV-1 variants and primary isolates. Recently, a crystal structure of the epitope bound to the Fab fragment of MAb 2F5 has shown that the 2F5 peptide adopts a beta-turn conformation [Pai, E. F., Klein, M. H., Chong, P., and Pedyczak, A. (2000) World Intellectual Property Organization Patent WO-00/61618]. We have designed cyclic peptides to adopt beta turn conformations by the incorporation of a side-chain to side-chain lactam bridge between the i and i + 4 residues containing the Asp-Lys-Trp segment. Synthesis of extended, nonconstrained peptides encompassing the 2F5 epitope revealed that the 13 amino acid sequence, Glu-Leu-Leu-Glu-Leu-Asp-Lys-Trp-Ala-Ser Leu-Trp-Asn, maximized MAb 2F5 binding. Constrained analogues of this sequence were explored to optimize 2F5 binding affinity. The solution conformations of the constrained peptides have been characterized by NMR spectroscopy and molecular modeling techniques. The results presented here demonstrate that both inclusion of the lactam constraint and extension of the 2F5 segment are necessary to elicit optimal antibody binding activity. The ability of these peptide immunogens to stimulate a high titer, peptide-specific immune response incapable of viral neutralization is discussed in regard to developing an HIV-1 vaccine designed to elicit a 2F5-like immune response. PMID- 12641453 TI - Understanding quinone cofactor biogenesis in methylamine dehydrogenase through novel cofactor generation. AB - Cofactors made from constitutive amino acids in proteins are now known to be relatively common. A number of these involve the generation of quinone cofactors, such as topaquinone in the copper-containing amine oxidases, and lysine tyrosylquinone in lysyl oxidase. The biogenesis of the quinone cofactor tryptophan tryptophylquinone (TTQ) in methylamine dehydrogenase (MADH) involves the post-translational modification of two constitutive Trp residues (Trp(beta)(57) and Trp(beta)(108) in Paracoccus denitrificans MADH). The modifications for generating TTQ are the addition of two oxygens to the indole ring of Trp(beta)(57) and the formation of a covalent cross-link between Cepsilon3 of Trp(beta)(57) and Cdelta1 of Trp(beta)(108). The order in which these events occur is unknown. To investigate the role Trp(beta)(108) may play in this process, this residue was mutated to both a His (betaW108H) and a Cys (betaW108C) residue. For each mutant, the majority of the protein that was isolated was inactive and exhibited weaker subunit-subunit interactions than native MADH. Analysis by mass spectrometry suggested that the inactive protein was a biosynthetic intermediate with only one oxygen atom incorporated into Trp(beta)(57) and no cross-link with residue beta108. However, in each mutant preparation, a small percentage of the mutant enzyme was active and appears to possess a functional tryptophylquinone cofactor. In the case of betaW108C, this cofactor may be identical to cysteine tryptophylquinone, recently described in the bacterial quinohemoprotein amine dehydrogenase. In betaW108H, the active cofactor is presumably a histidine tryptophylquinone, which has not been previously described, and represents the synthesis of a novel quinone protein cofactor. PMID- 12641454 TI - Oxidation of Met144 and Met145 in calmodulin blocks calmodulin dependent activation of the plasma membrane Ca-ATPase. AB - Methionine oxidation in calmodulin (CaM) isolated from senescent brain results in an inability to fully activate the plasma membrane (PM) Ca-ATPase, which may contribute to observed increases in cytosolic calcium levels under conditions of oxidative stress and biological aging. To identify the functional importance of the oxidation of Met(144) and Met(145) near the carboxyl-terminus of CaM, we have used site-directed mutagenesis to substitute leucines for methionines at other positions in CaM, permitting the site-specific oxidation of Met(144) and Met(145). Prior to their oxidation, the CaM-dependent activation of the PM-Ca ATPase by these CaM mutants is similar to that of wild-type CaM. Likewise, oxidation of individual methionines has a minimal effect on the CaM concentration necessary for half-maximal activation of the PM-Ca-ATPase. These results are consistent with previous suggestions that no single methionine within CaM is essential for activation of the PM-Ca-ATPase. Oxidation of either Met(144) and Met(145) or all nine methionines in CaM results in an equivalent inhibition of the PM-Ca-ATPase, resulting in a 50-60% reduction in the level of enzyme activation. Oxidation of Met(144) is largely responsible for the decreased extent of enzyme activation, suggesting that this site is critical in modulating the sensitivity of CaM to oxidant-induced loss-of-function. These results are discussed in terms of a possible functional role for Met(144) and Met(145) in CaM as redox sensors that function to modulate calcium homeostasis and energy metabolism in response to conditions of oxidative stress. PMID- 12641455 TI - Effects of vinylphosphonate internucleotide linkages on the cleavage specificity of exonuclease III and on the activity of DNA polymerase I. AB - We have previously reported the synthesis of vinylphosphonate-linked thymidine dimers and their incorporation into synthetic oligonucleotides to create vinylphosphonate internucleotide linkages in the DNA. Such linkages have a profound effect on DNA backbone rotational flexibility, and we have shown that the PcrA helicase, which requires such flexibility, is inhibited when it encounters these linkages on the translocating strand. In this study, we have investigated the effects of these linkages on the dsDNA specific exonuclease III and on the ssDNA specific mung bean nuclease to establish whether our modification confers resistance to nucleases making it suitable for antisense therapy applications. We also investigated the effect on DNA polymerase I to establish whether we could in the future use this enzyme to incorporate these linkages in the DNA. Our results show that a single modification does not affect the activity of DNA polymerase I, but four vinylphosphonate linkages in tandem inhibit its activity. Furthermore, such linkages do not confer significant nuclease resistance to either exonuclease III or mung bean nuclease, but unexpectedly, they alter the cleavage specificity of exonuclease III. PMID- 12641456 TI - Modulation of human nucleotide excision repair by 5-methylcytosines. AB - Previous reports showed that methylated CpG sites are primary targets of bulky lesions induced by UV radiation, benzo[a]pyrene (B[a]P), or other environmental genotoxic agents. This study was performed to determine whether the repair of DNA damage formed preferentially at CpG dinucleotides is sensitive to 5 methylcytosine substitutions. Reactivation assays using UV- or B[a]P diol epoxide damaged shuttle vectors established that human nucleotide excision repair enzymes are able to process fully methylated target DNA molecules. Repair reactions in human cell extracts suggested that 5-methylcytosines modulate local repair efficiency in a seemingly unpredictable manner. In fact, excision of the predominant (+)-trans-anti-B[a]P-dG adduct situated in a mutational hot spot sequence (codon 273 of the p53 gene) was stimulated by CpG methylation. Interestingly, excision activity was increased by a single 5-methylcytosine residue flanking the adduct in the damaged strand, but the same stimulatory effect was also induced by a single 5-methylcytosine residue located opposite the adduct in the undamaged strand. No such stimulation was observed when the (+) trans-anti-B[a]P-dG lesion was placed in a different site containing a sequence of contiguous guanines, and strong inhibition was detected when a representative of the rare (+)-cis-anti-B[a]P-dG isomer was tested in the same assay. These results raise the possibility that 5-methylcytosines in CpG dinucleotides modulate not only the distribution of bulky DNA lesions but, at least in some cases, also the kinetics of subsequent excision repair reactions. This study confirms that the efficiency of bulky lesion repair is determined by the configuration of base pairs at damaged sites. PMID- 12641457 TI - RPA phosphorylation in mitosis alters DNA binding and protein-protein interactions. AB - The heterotrimeric DNA-binding protein, replication protein A (RPA), consists of 70-, 34-, and 14-kDa subunits and is involved in maintaining genomic stability by playing key roles in DNA replication, repair, and recombination. RPA participates in these processes through its interaction with other proteins and its strong affinity for single-stranded DNA (ssDNA). RPA-p34 is phosphorylated in a cell cycle-dependent fashion primarily at Ser-29 and Ser-23, which are consensus sites for Cdc2 cyclin-dependent kinase. By systematically examining RPA-p34 phosphorylation throughout the cell cycle, we have found there are distinct phosphorylated forms of RPA-p34 in different cell-cycle stages. We have isolated and purified a unique phosphorylated form of RPA that is specifically associated with the mitotic phase of the cell cycle. The mitotic form of RPA (m-hRPA) shows no difference in ssDNA binding activity as compared with recombinant RPA (r hRPA), yet binds less efficiently to double-stranded DNA (dsDNA). These data suggest that mitotic phosphorylation of RPA-p34 inhibits the destabilization of dsDNA by RPA complex, thereby decreasing the binding affinity for dsDNA. The m hRPA also exhibits altered interactions with certain DNA replication and repair proteins. Using highly purified proteins, m-hRPA exhibited decreased binding to ATM, DNA pol alpha, and DNA-PK as compared to unphosphorylated recombinant RPA (r hRPA). Dephosphorylation of m-hRPA was able to restore the interaction with each of these proteins. Interestingly, the interaction of RPA with XPA was not altered by RPA phosphorylation. These data suggest that phosphorylation of RPA-p34 plays an important role in regulating RPA functions in DNA metabolism by altering specific protein-protein interactions. PMID- 12641458 TI - Using a novel dual fluorescence quenching assay for measurement of tryptophan depth within lipid bilayers to determine hydrophobic alpha-helix locations within membranes. AB - A novel fluorescence method for determining the depth of Trp residues in membrane inserted polypeptides is introduced. Quenching of Trp by acrylamide and 10 doxylnonadecane (10-DN) was used to measure Trp depth. Transmembrane helices with Trp residues at varying positions (and thus locating at different depths in lipid bilayers) were used to calibrate the method. It was found that acrylamide quenches Trp close to the bilayer surface more strongly than it quenches deeply buried Trp, while 10-DN quenches Trp close to the center of the bilayer more strongly than Trp close to the surface. The ratio of acrylamide quenching to that of 10-DN was found to be nearly linearly dependent on the depth of Trp in a membrane. It was also found that it was possible to detect coexisting shallowly and deeply inserted populations of Trp-containing polypeptides using these quenchers. In the presence of such mixed populations, acrylamide induced large blue shifts in fluorescence emission lambda(max) whereas 10-DN induced large red shifts. In a more homogeneous population quencher-induced shifts were found to be minimal. Dual quencher analysis can be used to distinguish hydrophobic helices with a transmembrane orientation from those located close to the bilayer surface and, when applied to a number of different peptides, revealed novel aspects of hydrophobic helix behavior. PMID- 12641459 TI - Cumulative effects of amino acid substitutions and hydrophobic mismatch upon the transmembrane stability and conformation of hydrophobic alpha-helices. AB - The effects of amino acid substitutions upon the behavior of poly(Leu)-rich alpha helices inserted into model membrane vesicles were investigated. One or two consecutive Leu residues in the hydrophobic core of the helix were substituted with A, F, G, S, D, K, H, P, GG, SS, PG, PP, KK, or DD residues. A Trp placed at the center of the sequence allowed assessment of peptide behavior via fluorescence emission lambda(max) and dual quenching analysis of Trp depth [Caputo, G. A., and London, E. (2003) Biochemistry 42, 3265-3274]. In vesicles composed of dioleoylphosphatidylcholine (DOPC), all of the peptides with single substitutions adopted a transmembrane (TM) state. Experiments were also performed in thicker bilayers composed of dierucoylphosphatidylcholine (DEuPC). In DEuPC vesicles TM states were destabilized by mismatch between helix length and bilayer thickness. Nevertheless, in DEuPC vesicles TM states were still prevalent for peptides with single substitutions, although less so for peptides with P, K, H, or D substitutions. In contrast to single substitutions, certain consecutive double substitutions strongly interfered with formation of TM states. In both DOPC and DEuPC vesicles DD and KK substitutions abolished the normal TM state, but GG and SS substitutions had little effect. In even wider bilayers, a SS substitution reduced the formation of a TM state. A peptide with a PP substitution maintained the TM state in DOPC vesicles, but in DEuPC vesicles the level of formation of the TM state was significantly reduced. Upon disruption of normal TM insertion peptides moved close to the bilayer surface, with the exception of the KK-substituted peptide in DOPC vesicles, which formed a truncated TM segment. These studies begin to provide a detailed relationship between sequence and the stability of TM insertion and show that the influence of insertion-destabilizing residues upon hydrophobic helices can be strongly modulated by properties such as mismatch. For certain helix-forming hydrophobic sequences, sensitivity to lipid structure may be sufficient to induce large conformational changes in vivo. PMID- 12641460 TI - Binding of a photoaffinity analogue of glutathione to MRP1 (ABCC1) within two cytoplasmic regions (L0 and L1) as well as transmembrane domains 10-11 and 16-17. AB - MRP1 (or ABCC1) is an ABC membrane protein that transports a wide range of natural products as well as glutathione (GSH)-, glucuronate-, and sulfate conjugated metabolites. In addition, free GSH is required for MRP1 to transport several chemotherapeutic drugs. However, the mechanisms regulating the influence of GSH on MRP1 is poorly understood, and the location of GSH binding site(s) within MRP1 have yet to be determined. To address these issues, we have synthesized a [(125)I] labeled azido-derivative of GSH (IAAGSH) to photoaffinity label MRP1. Our results revealed that IAAGSH labeled MRP1 with high specificity, and binding was inhibited by MRP1 substrates leukotriene C(4) and MK571. Interestingly, verapamil and vincristine enhanced IAAGSH photolabeling of MRP1, in agreement with observations that both drugs enhance GSH transport. We observed GSH to be the best inhibitor of photoaffinity labeling, as compared to oxidized glutathione (GSSG) and four different GSH alkyl derivatives. These observations indicate that IAAGSH interacted with MRP1 in a similar manner as unmodified GSH. Moreover, using eight MRP1-HA variants, each containing hemagglutinin A (HA) epitopes inserted at different sites in MRP1, we mapped the GSH binding sites in MRP1. Our GSH analogue photoaffinity labeled four MRP1 polypeptides that were located within two cytoplasmic domains in linker sequences (L0 and L1) as well as transmembrane domains 10-11 and 16-17. The photoaffinity labeling of polypeptides within L0 and L1 domains is further confirmed using two MRP1-specific monoclonal antibodies (MRPr1 and QCRL1) with epitopes within the linker domains. Taken together, this study provides the most precise information to date on the location of GSH binding sites in MRP1. PMID- 12641461 TI - Structural changes of the prion protein in lipid membranes leading to aggregation and fibrillization. AB - Prion diseases are associated with a major refolding event of the normal cellular prion protein, PrP(C), where the predominantly alpha-helical and random coil structure of PrP(C) is converted into a beta-sheet-rich aggregated form, PrP(Sc). Under normal physiological conditions PrP(C) is attached to the outer leaflet of the plasma membrane via a GPI anchor, and it is plausible that an interaction between PrP and lipid membranes could be involved in the conversion of PrP(C) into PrP(Sc). Recombinant PrP can be refolded into an alpha-helical structure, designated alpha-PrP isoform, or into beta-sheet-rich states, designated beta-PrP isoform. The current study investigates the binding of beta-PrP to model lipid membranes and compares the structural changes in alpha- and beta-PrP induced upon membrane binding. beta-PrP binds to negatively charged POPG membranes and to raft membranes composed of DPPC, cholesterol, and sphingomyelin. Binding of beta-PrP to raft membranes results in substantial unfolding of beta-PrP. This membrane associated largely unfolded state of PrP is slowly converted into fibrils. In contrast, beta-PrP and alpha-PrP gain structure with POPG membranes, which instead leads to amorphous aggregates. Furthermore, binding of beta-PrP to POPG has a disruptive effect on the integrity of the lipid bilayer, leading to total release of vesicle contents, whereas raft vesicles are not destabilized upon binding of beta-PrP. PMID- 12641462 TI - Mutation in rod PDE6 linked to congenital stationary night blindness impairs the enzyme inhibition by its gamma-subunit. AB - Photoreceptor cGMP phosphodiesterase (PDE6) is the effector enzyme in the vertebrate visual transduction cascade. The activity of rod PDE6 catalytic alpha- and beta-subunits is blocked in the dark by two inhibitory Pgamma-subunits. The inhibition is released upon light-stimulation of photoreceptor cells. Mutation H258N in PDE6beta has been linked to congenital stationary night blindness (CSNB) in a large Danish family (Rambusch pedigree) (Gal, A., Orth, U., Baehr, W., Schwinger, E., and Rosenberg, T. (1994) Nat. Genet. 7, 64-67.) We have analyzed the consequences of this mutation for PDE6 function using a Pgamma-sensitive PDE6alpha'/PDE5 chimera, Chi16. Biochemical analysis of the H257N mutant, an equivalent of PDE6betaH258N, demonstrates that this substitution does not alter the ability of chimeric PDE to dimerize or the enzyme's catalytic properties. The sensitivity of H257N to a competitive inhibitor zaprinast was also unaffected. However, the mutant displayed a significant impairment in the inhibitory interaction with Pgamma, which was apparent from a approximately 20-fold increase in the K(i) value (46 nM) and incomplete maximal inhibition. The inhibitory defect of H257N is not due to perturbation of noncatalytic cGMP binding to the PDE6alpha' GAF domains. The noncatalytic cGMP-binding characteristics of the H257N mutant were similar to those of the parent PDE6alpha'/PDE5 chimera. Since rod PDE6 in the Rambusch CSNB is a catalytic heterodimer of the wild-type PDE6alpha and mutant PDE6beta, Chi16 and H257N were coexpressed, and a heterodimeric PDE, Chi16/H257N, was isolated. It displayed two Pgamma inhibitory sites with the K(i) values of 5 and 57 nM. Our results support the hypothesis that mutation H258N in PDE6beta causes CSNB through incomplete inhibition of PDE6 activity by Pgamma, which leads to desensitization of rod photoreceptors. PMID- 12641463 TI - Control of the reactivation kinetics of homodimeric triosephosphate isomerase from unfolded monomers. AB - Homodimeric triosephosphate isomerases from Trypanosoma cruzi (TcTIM) and Trypanosoma brucei (TbTIM) have markedly similar catalytic properties and 3-D structures; their overall amino acid sequence identity is 68% and 85% in their interface residues. Nonetheless, active dimer formation from guanidinium chloride unfolded monomers is faster and more efficient in TcTIM than in TbTIM. The enzymes thus provide a unique opportunity for exploring the factors that control the formation of active dimers. The kinetics of reactivation at different protein concentrations showed that the process involved three reactions: monomer folding, association of folded monomers, and a transition from inactive to active dimers. The rate constants of the reactions indicated that, at relatively low protein concentrations, the rate-limiting step of reactivation was the association reaction; at high protein concentrations the transition of inactive to active dimers was rate limiting. The rates of the latter two reactions were higher in TcTIM than in TbTIM. Studies with a mutant of TcTIM that had the interface residues of TbTIM showed that the association rate constant was similar to that of TbTIM. However, the rate of the transition from inactive to active dimers was close to that of TcTIM; thus, this transition depends on the noninterfacial portion of the enzymes. When unfolded monomers of TcTIM and TbTIM were allowed to reactivate together, TcTIM, the hybrid, and TbTIM were formed in a proportion of 1:0.9:0.2. This distribution suggests that, in the hybrid, the characteristics of the TcTIM monomers influence the properties of TbTIM monomers. PMID- 12641464 TI - Site-specific mutations provide new insights into the origin of pH effects and alternative spectral forms in the photoactive yellow protein from Halorhodospira halophila. AB - Acid/base titrations of wild-type PYP and mutants, either in buffer or in the presence of chaotropes such as thiocyanate, establish the presence of four spectral forms including the following: a neutral form (446-476 nm), an acidic form (350-355 nm), an alkaline form (430-440 nm), and an intermediate wavelength form (355-400 nm). The acidic species is formed by protonation of the oxyanion of the para-hydroxy-cinnamyl cysteine chromophore as a secondary result of acid denaturation (with pK(a) values of 2.8-5.4) and often results in precipitation of the protein, and in the case of wild-type PYP, eventual hydrolysis of the chromophore thioester bond at pH values below 2. Thus, the large and complex structural changes associated with the acidic species make it a poor model for the long-lived photocycle intermediate, I(2), which undergoes more moderate structural changes. Mutations at E46, which is hydrogen-bonded to the chromophore, have only two spectral forms accessible to them, the neutral and the acidic forms. Thus, an intact E46 carboxyl group is essential for observation of either intermediate or alkaline wavelength forms. The alkaline form is likely to be due to ionization of E46 in the folded protein. We postulate that the intermediate wavelength form is due to a conformational change that allows solvent access to E46 and formation of a hydrogen-bond from a water molecule to the carboxylic acid group, thus weakening its interaction with the chromophore. Increasing solvent access to the intermediate spectral form with denaturant concentration results in a continuously blue-shifted wavelength maximum. PMID- 12641465 TI - Mechanism of S-nitrosation of recombinant human brain calbindin D28K. AB - Mass spectrometry and UV-vis absorption results support a mechanism for NO donation by S-nitrosoglutathione (GSNO) to recombinant human brain calbindin D(28K) (rHCaBP) that requires the presence of trace copper, added as either Cu,Zn superoxide dismutase (CuZnSOD) or CuSO(4). The extent of copper-catalyzed rHCaBP S-nitrosation depends on the ratio of protein to GSNO and on the reaction time, and NO-transfer is prevented when copper chelators are present. CuZnSOD is an efficient catalyst of rHCaBP S-nitrosation, and the mechanism of CuZnSOD catalyzed S-nitrosation involves reduction of the active-site Cu(II) by a number of the five free thiols in rHCaBP, giving rise to thiyl radicals. The Cu(I)ZnSOD formed catalyzes the reductive cleavage of GSNO present in solution to give GSH and release NO. rHCaBP thiyl radicals react with NO to yield the S nitrosoprotein. Cu(II)ZnSOD is also reduced by GSH in a concentration-dependent manner up to 5 mM but not at higher GSH concentrations. However, unlike the rHCaBP thiyl radicals, GS(*) radicals dimerize to GSSG faster than their reaction with NO. The data presented here provide a biologically relevant mechanism for protein S-nitrosation by small S-nitrosothiols. S-nitrosation is rapidly gaining recognition as a major form of protein posttranslational modification, and the efficient S-nitrosation of CaBP by CuZnSOD/GSNO is speculated to be of neurochemical importance given that CaBP and CuZnSOD are abundant in neurons. PMID- 12641466 TI - Mechanistic origins of the substrate selectivity of serine proteases. AB - Serine proteases catalyze the hydrolysis of amide bonds of their protein and peptide substrates through a mechanism involving the intermediacy of an acyl enzyme. While the rate constant for formation of this intermediate, k(2), shows a dramatic dependence on peptide chain length, the rate constant for the intermediate's hydrolysis is relatively insensitive to chain length. To probe the mechanistic origins of this phenomenon, we determined temperature dependencies and solvent isotope effects for the alpha-chymotrypsin-catalyzed hydrolysis of Suc-Phe-pNA (K(s) = 1 mM, k(2) = 0.04 s(-)(1), and k(3) = 11 s(-)(1)), Suc-Ala Phe-pNA (K(s) = 4 mM, k(2) = 0.9 s(-)(1), and k(3) = 42 s(-)(1)), and Suc-Ala-Ala Pro-Phe-pNA (K(s) = 0.1 mM, k(2) = 98 s(-)(1), and k(3) = 71 s(-)(1)). We found that while the van't Hoff plots for K(s) and the Eyring plots for k(3) are linear for all three reactions, the Eyring plots for k(2) are convex, indicating that the process governed by k(2) is complex, possibly involving a coupling between active site chemistry and protein conformational isomerization. This interpretation is strengthened by solvent isotope effects on k(2) that are largely temperature-independent. Furthermore, the dependence of k(2) on peptide length is manifested entirely in the enthalpy of activation, suggesting a mechanism of catalysis by distortion. Taken together, this analysis of acylation suggests that extended substrates which can engage in subsite interactions are able to efficiently trigger the coupling mechanism between chemistry and a conformational isomerization that distorts the substrate and thereby promotes nucleophilic attack. PMID- 12641467 TI - DSC studies of a family of natively disordered fragments from Escherichia coli thioredoxin: surface burial in intrinsic coils. AB - The accumulating data from proteome analysis indicates that numerous proteins have segments and/or domains, involved in regulatory functions of the eukaryotic cell, which are entirely unstructured under physiological conditions, challenging the structure-function paradigm. Although many such natively unfolded proteins have been structurally analyzed by NMR spectroscopy, little is known about solvent inaccessible surfaces in premolten globules and intrinsic coils. Recent DSC studies of two protein fragments have shown a promising way to estimate the predominantly hydrophobic buried surfaces [Georgescu, R. E., Garcia-Mira, M. M., Tasayco, M. L., and Sanchez-Ruiz, J. M. (2001) Eur. J. Biochem. 268, 1-10]. Here we report a systematic heat capacity analysis of a family of natively disordered complementary fragments of oxidized Escherichia coli thioredoxin (1-31/32-108, 1 37/38-108, 1-50/51-108, and 38-73) which provides insights into the local and nonlocal interactions contributing to the burial of predominantly hydrophobic surface in intrinsic coils. PMID- 12641468 TI - Bacterial phytoene synthase: molecular cloning, expression, and characterization of Erwinia herbicola phytoene synthase. AB - Phytoene synthase (PSase) catalyzes the condensation of two molecules of geranylgeranyl diphosphate (GGPP) to give prephytoene diphosphate (PPPP) and the subsequent rearrangement of the cyclopropylcarbinyl intermediate to phytoene. These reactions constitute the first pathway specific step in carotenoid biosynthesis. The crtB gene encoding phytoene synthase was isolated from a plasmid containing the carotenoid gene cluster in Erwinia herbicola and cloned into an Escherichia coli expression system. Upon induction, recombinant phytoene synthase constituted 5-10% of total soluble protein. To facilitate purification of the recombinant enzyme, the structural gene for PSase was modified by site directed mutagenesis to incorporate a C-terminal Glu-Glu-Phe (EEF) tripepetide to allow purification by immunoaffinity chromatography on an immobilized monoclonal anti-alpha-tubulin antibody YL1/2 column. Purified recombinant PSase-EEF gave a band at 34.5 kDa upon SDS-PAGE. Recombinant PSase-EEF was then purified to >90% homogeneity in two steps by ion-exchange and immunoaffinity chromatography. The enzyme required Mn(2+) for activity, had a pH optimum of 8.2, and was strongly stimulated by detergent. The concentration of GGPP needed for half-maximal activity was approximately 35 microM, and a significant inhibition of activity was seen at GGPP concentrations above 100 microM. The sole product of the reaction was 15,15'-Z-phytoene. PMID- 12641469 TI - Dispensability of glutamic acid 48 and aspartic acid 134 for Mn2+-dependent activity of Escherichia coli ribonuclease HI. AB - The activities of the eight mutant proteins of Escherichia coli RNase HI, in which the four carboxylic amino acids (Asp(10), Glu(48), Asp(70), and Asp(134)) involved in catalysis are changed to Asn (Gln) or Ala, were examined in the presence of Mn(2+). Of these proteins, the E48A, E48Q, D134A, and D134N proteins exhibited the activity, indicating that Glu(48) and Asp(134) are dispensable for Mn(2+)-dependent activity. The maximal activities of the E48A and D134A proteins were comparable to that of the wild-type protein. However, unlike the wild-type protein, these mutant proteins exhibited the maximal activities in the presence of >100 microM MnCl(2), and their activities were not inhibited at higher Mn(2+) concentrations (up to 10 mM). The wild-type protein contains two Mn(2+) binding sites and is activated upon binding of one Mn(2+) ion at site 1 at low ( approximately 1 microM) Mn(2+) concentrations. This activity is attenuated upon binding of a second Mn(2+) ion at site 2 at high (>10 microM) Mn(2+) concentrations. The cleavage specificities of the mutant proteins, which were examined using oligomeric substrates at high Mn(2+) concentrations, were identical to that of the wild-type protein at low Mn(2+) concentrations but were different from that of the wild-type protein at high Mn(2+) concentrations. These results suggest that one Mn(2+) ion binds to the E48A, E48Q, D134A, and D134N proteins at site 1 or a nearby site with weaker affinities. The binding analyses of the Mn(2+) ion to these proteins in the absence of the substrate support this hypothesis. When Mn(2+) ion is used as a metal cofactor, the Mn(2+) ion itself, instead of Glu(48) and Asp(134), probably holds water molecules required for activity. PMID- 12641470 TI - Evaluation of number average length analysis in quantifying double strand breaks in genomic DNAs. AB - Double strand breaks in DNA can be quantified down to very low frequencies (a few per Gigabase pair) in nanogram quantities of nonradioactive, genomic DNA by dispersing the DNAs on electrophoretic gels, digitizing them by quantitative electronic imaging, and calculating the DNA lengths by number average length analysis. No specific distribution of damages is required for number average length analysis. To test the validity of this approach, we used DNA populations of known absolute lengths and break frequencies as experimental DNAs and calculated the number average lengths and double strand break levels. Experimental DNAs and length standards were dispersed using pulsed field electrophoretic modes (unidirectional pulsed field, contour clamped homogeneous field, or transverse alternating field) appropriate for their size range, stained with ethidium, destained, and a quantitative electronic image obtained. A dispersion curve was constructed from the migration-mobility relationships of the length standard DNAs, and the number average lengths of the experimental DNAs were calculated. The calculated DNA lengths agreed well with the actual lengths. Furthermore, the double strand break frequencies calculated through number average length analysis of DNAs dispersed by these pulsed field gel modes and digitized by quantitative electronic imaging were in excellent agreement with the actual values for populations of DNA over the size range of approximately 4 kbp to approximately 3 Mbp. The use of this approach in quantifying DNA damages is illustrated for double strand breaks and damage clusters (e.g., OxyPurine clusters recognized by Escherichia coli Fpg protein) induced in T7 DNA by ionizing radiation. PMID- 12641471 TI - Structural genomics. PMID- 12641472 TI - Facilities and methods for the high-throughput crystal structural analysis of human proteins. AB - Facilities and methods for the high-throughput crystal structure analysis of human proteins are described as recently established in the Protein Structure Factory, a Berlin-area structural genomics project. Genes encoding human proteins are expressed in either recombinant Escherichia coli or yeast (Saccharomyces cerevisiae or Pichia pastoris). To facilitate and standardize protein purification, the target proteins are produced with various tags for affinity chromatography. For high-throughput crystallization, a robotic station is being set up that has the capacity to handle 960 000 experiments simultaneously. The resulting protein crystals will be subjected to X-ray diffraction experiments at the third-generation synchrotron storage ring BESSY where protein crystallography beamlines are currently under construction. The Protein Structure Factory's strategy for high-throughput production and structure analysis of human proteins is evaluated based on first results. PMID- 12641473 TI - Medium-scale structural genomics: strategies for protein expression and crystallization. AB - While high-throughput methods of protein production and crystallization are beginning to be well documented, owing to the output of large structural genomics programs, medium-throughput methods at the laboratory scale lag behind. In this paper, we report a possible way for an academic laboratory to adapt high throughput to medium-throughput methods, on the basis of the first results of two projects aimed at solving the 3D structures of Escherichia coli and Mycobacterium tuberculosis (Tb) proteins of unknown function. We have developed sequential and iterative procedures as well as new technical processes for these programs. Our results clearly demonstrate the value of this medium-throughput approach. For instance, in the first 14 months of the E. coli program, 69 out of 108 target genes led to soluble proteins, 36 were brought to crystallization, and 28 yielded crystals; among the latter, 13 led to usable data sets and 9 to structures. These results, still incomplete, might help in planning future directions of expression and crystallization of proteins applied to medium-throughput structural genomics programs. PMID- 12641474 TI - High-throughput crystallography at an affordable cost: the TB Structural Genomics Consortium Crystallization Facility. AB - The crystallization facility of the TB Structural Genomics Consortium, one of nine P50 structural genomics centers sponsored by the National Institutes of Health, provides TB consortium members with automated crystallization, data collection, and basic molecular replacement structure solution up to bias minimized maps. In contrast to venture capital -funded commercial enterprises, the TB consortium facilities are decentralized and aim to develop high-throughput crystallography methods and automation on a comparatively small budget. In addition to financial constraints, the logistics and organization of a production environment differ considerably from academic settings. The TB Structural Genomics Consortium crystallization facility may thus provide a model for cost effective, efficient high-throughput crystallography. Processes and methods presented in this review should assist academic institutions planning to invest in high-throughput structural biology to assess both the rewards and risks of establishing structural genomics programs. PMID- 12641475 TI - Structural proteomics: toward high-throughput structural biology as a tool in functional genomics. AB - Structural proteomics is the determination of atomic resolution three-dimensional protein structures on a genome-wide scale in order to better understand the relationship between protein sequence, structure, and function. Here we describe our ongoing structural proteomics project on the nonmembrane proteins of the archeaon, Methanobacterium thermoautotrophicum. This article provides a snapshot of an ongoing pilot project in an emerging area of multidisciplinary research that involves bioinformatics, molecular biology, biochemistry, and instrumental methods such as NMR spectroscopy and X-ray crystallography. An assessment of the technical challenges in this type of large-scale project along with a comparison of the efficiency of sample production for both X-ray crystallography and NMR spectroscopy will be discussed. Examples of new insights into protein function and the relationship between structure and sequence will also be presented. PMID- 12641476 TI - The Southeast Collaboratory for Structural Genomics: a high-throughput gene to structure factory. AB - The Southeast Collaboratory for Structural Genomics consists of four working groups. The protein production group supplies/develops high-output production of Pyrococcus furiosus, Caenorhabditis elegans, and selected human proteins. The X ray crystallography group conducts high-throughput structure production in parallel with production-related research/development in nanocrystallization robotics, capillary crystallization cassette, synchrotron/home X-ray instrumentation, sample mounting robotics, data processing and pipelined structure analysis, combined refinement/validation protocols, and direct use of unlabeled native crystals (Direct Crystallography). The NMR group emphasizes/develops sample screening and backbone structure determination from residual dipolar coupling data. The bioinformatics group implements/develops local database interfaces, pipelined sequence/structure information search/updates, and database/bioinformatics toolkits. PMID- 12641477 TI - Structural genomics of membrane proteins. AB - A program on the structural genomics of membrane proteins has started at the BIRC, AIST, involving other academic institutions and industrial companies. Emphasis is being put on the development of techniques for the structural determination of membrane proteins of biological importance and ligand-receptor interactions by means of electron microscopy, X-ray diffraction, NMR, and computer simulation. Most efforts at the present stage, however, are being directed to finding suitable expression and purification systems and crystallization conditions for such proteins. The program is expected to be linked with the human full-length cDNA project and should lead to medical and industrial uses. PMID- 12641478 TI - NMR and structural genomics. AB - The role of NMR in structural genomics is outlined, with particular emphasis on using protein domains as targets. Strategies for domain expression, characterization, and labeling are presented. PMID- 12641479 TI - Structural genomics of proteins involved in copper homeostasis. AB - Genome sequencing projects have provided a wealth of data, most notably the primary sequences of all the proteins that a given organism can produce. The understanding of this information at the functional level is still in the beginning stages. Three-dimensional structural information is necessary to unravel at the atomic level the mechanisms by which a protein carries out its function, and such information can often be very useful to predict at least gross functional features, even in the absence of biochemical data. An exhaustive structural characterization of the proteins encoded in the genomes is thus highly desirable. To enhance the functional insights provided by genome-scale structural determination, we have prioritized our research to target specific processes of the cell, i.e., those responsible for controlling metal homeostasis. In this Account, we present the results obtained by the Magnetic Resonance Center of the University of Florence on proteins involved in the homeostasis of copper. The general research strategy is presented, followed by a discussion focused on different key experimental aspects. An overview of the initial results and of their relevance to the understanding of molecular function and cellular processes is also given. PMID- 12641480 TI - The Perindopril Protection Against Recurrent Stroke Study (PROGRESS): clinical implications for older patients with cerebrovascular disease. AB - Blood pressure levels are strongly predictive of the risks of first-ever and recurrent stroke. The benefits of blood pressure-lowering therapy for the prevention of fatal and non-fatal stroke in middle-aged individuals are well established. However, until recently, there has been uncertainty about the consistency of such benefits across different patient groups and in particular, for older people and in those with a history of stroke. This paper discusses the evidence surrounding the effectiveness of blood pressure-lowering therapy, specifically in older patients with a history of stroke, with particular attention paid to the results from the Perindopril Protection Against Recurrent Stroke Study (PROGRESS). PROGRESS was a randomised, double-blind, placebo controlled trial of 6105 individuals with a history of cerebrovascular disease recruited from 172 hospital outpatient clinics in ten countries. Participants (mean age 64 years; range 26-91 years) were randomly assigned to receive active treatment with an ACE inhibitor-based blood pressure-lowering regimen (perindopril) with or without addition of the diuretic indapamide, or matched placebo. At the end of follow up (mean of 4 years), active treatment reduced the incidence of total stroke by 28% (95% CI 17-38%) and the rate of major vascular events by 26% (95% CI 16-34%). Importantly, benefits of treatment were consistent across key patient subgroups, including those with and without hypertension, patients who were Asian and non-Asian, and for both ischaemic and haemorrhagic strokes subtypes. Current evidence is now strong for clinicians to consider blood pressure-lowering therapy as pivotal in the prevention of stroke, especially in patients with a known history of cerebrovascular disease (and vascular disease, in general), irrespective of blood pressure levels, as soon as patients are clinically stable after an acute stroke or other vascular event. Additional age specific analyses of the PROGRESS data, together with those from other completed trials, will provide more reliable information about the size of the benefits of blood pressure-lowering therapy, specifically for different age groups, and particularly in the oldest old (those aged >80 years). In the meantime though, an ACE inhibitor plus diuretic treatment regimen that maximises the degree of blood pressure reduction has a good safety profile and is an effective treatment that should be considered in all patients with stroke, including the elderly. PMID- 12641481 TI - Cost of pharmacological care of the elderly: implications for healthcare resources. AB - Expenditures on prescribed medicines are significantly higher among those aged 65 years and over than among younger people. As populations in developed countries age so the cost of pharmacological care associated with the older population can be expected to increase. While pharmacological care represents only one component of healthcare, its costs are increasing rapidly because of advances in technology and increasing use. However, such costs should be considered within a context of decreasing disability in the elderly population, improving economic conditions among seniors and the relationship of these costs with those in other aspects of healthcare. Where medications have been demonstrated to be cost-effective, attempts to curtail expenditure growth may prove a false economy resulting in significantly higher growth elsewhere such as in the hospital and long-term care sectors. Policy responses to this issue should encompass the inclusion of elderly patients in clinical trials, the use of evidence-based principles of practice and strategies to ensure that this population obtain maximum benefit from medication through education and counselling. PMID- 12641483 TI - Prevalence and implications of uncontrolled systolic hypertension. AB - Risk of cardiovascular disease (CVD) increases incrementally with blood pressure, even within the high-normal range. In the general population, 27% of CVD in women and 37% in men is attributable to hypertension. A high percentage of these hypertension-related events occur in those with high-normal blood pressure and mild hypertension; about one-fourth of CVD events in elderly women and one-third in elderly men in the Framingham Study occurred in persons who had blood pressures of 140-159 mm Hg systolic and/or 90-95 mm Hg diastolic. The average systolic blood pressure (SBP) at which coronary heart disease occurs is rather modest (141 mm Hg), as is the pulse pressure (59-63 mm Hg). Of the CVD events in elderly participants in the Framingham Study, 24% in men and 36% in women occurred in persons receiving treatment for hypertension. There is a growing recognition of the importance of the systolic component of blood pressure. About 65% of hypertension in the elderly is isolated systolic hypertension (ISH), and CVD risk increases with pulse pressure. Pulse pressure is not simply a marker for stiff diseased arteries; treatment of ISH in trials promptly reduces the CVD risk, indicating that the pulse pressure generated by the stiff artery is the culprit. Analysis of data from clinical trials indicates that greater reliance should be placed on systolic pressure in evaluating the CVD potential of hypertension. Hypertension, including ISH, seldom occurs in isolation from other risk factors and overt CVD. Risk varies widely depending on the burden of accompanying risk factors. This makes global risk assessment mandatory for evaluating risk and the urgency and nature of treatment required. Evidence incriminating systolic pressure as the dominant blood pressure determinant of CVD has not been translated into clinical practice. Most of the uncontrolled hypertension observed in the Framingham Study is concentrated in those with ISH. This also extends to African-Americans, people with diabetes mellitus and the elderly. When should SBP be considered controlled? Substantial evidence supports the value of treating ISH with SBP exceeding 160 mm Hg. Trial data are not yet available to support recommendations to treat lesser elevations of ISH or pulse pressure per se, but since one-half of patients with mild ISH have two or more additional risk factors, most are candidates for treatment. In such patients, ISH should be considered controlled when their global CVD risk is reduced to below the average for their age. PMID- 12641484 TI - Systolic blood pressure elevation: it's where the action is. PMID- 12641482 TI - Statin therapy in the elderly: does it make good clinical and economic sense? AB - HMG-CoA reductase inhibitors (statins) have been established as the dominant treatment for coronary heart disease (CHD). This dominance is based on an impressive body of clinical trial evidence showing significant benefits in primary prevention of cardiovascular events in individuals at risk for CHD and in secondary prevention of such events in patients with CHD and high or normal plasma cholesterol levels. There is, however, significant room for improvement in the treatment of CHD with respect both to drug efficacy and to the disparity between evidence-based medicine and actual clinical practice particularly in relation to treatment strategies for the elderly. Current statins fall short of requirements for 'ideal' lipid-lowering treatment in several respects; 'super' statins and other agents currently in development may satisfy more of these requirements. Moreover, available therapies are not applied optimally, because of physician nonacceptance and/or patient noncompliance; thus, the majority of patients with CHD or its risk factors still have cholesterol levels that exceed guideline targets. There is also evidence that older patients with CHD, or at high risk of CHD, are undertreated - possibly because of concerns regarding the increased likelihood of adverse events or drug interactions or doubts regarding the cost effectiveness of statin therapy in this population. This group is of particular clinical relevance, since it is showing a proportionate rapid expansion in most national populations. To address their potential healthcare needs, the ongoing Pravastatin in the Elderly at Risk (PROSPER) study is assessing the effects of pravastatin in elderly patients (5804 men and women aged 70-82 years) who either have pre-existing vascular disease or are at significant risk for developing it, with the central hypothesis that statin therapy (pravastatin 40 mg/day) will diminish the risk of subsequent major vascular events compared with placebo. After a 3.2-year treatment period, a primary assessment will be made of the influence of statin treatment on major cardiovascular events (a combination of CHD death, nonfatal myocardial infarction, and fatal or nonfatal stroke). Optimal deployment of the currently available agents and of newer agents (no matter how well they satisfy requirements for ideal treatment) ultimately depends on the establishment of an evidence base and may require far-reaching educational programmes that change the way risk factor management is viewed by caregivers and patients alike. PMID- 12641485 TI - Fluoroquinolones in the elderly: safety considerations. AB - Fluoroquinolones such as ciprofloxacin, levofloxacin, moxifloxacin and gatifloxacin are widely used for the treatment of bacterial infections. Fluoroquinolone-induced adverse effects have not been reported to occur with increased frequency in the elderly, but large trials comparing the tolerability in aged and young individuals are not available. Renal function declines consistently with age and recommendations for dosage changes of renally eliminated fluoroquinolones (ofloxacin, levofloxacin, gatifloxacin) are related to changes in kidney function rather than to age per se. However, during routine clinical work, creatinine clearance data are usually not available; thus it seems more practical to recommend dosage adjustment for elderly individuals in whom low creatinine clearance values can be expected. Reactions of the gastrointestinal tract are the most often observed adverse effects during therapy with fluoroquinolones; however, compared with many other antibacterials, fluoroquinolones are less frequently associated with diarrhoea. Similarly, hypersensitivity reactions, as observed during therapy with penicillins and other beta-lactam agents, occur more rarely during fluoroquinolone therapy. Adverse reactions of the CNS are of particular concern for the elderly population. Elderly patients with impairments of the CNS (e.g. epilepsy, pronounced arteriosclerosis) should be treated with fluoroquinolones only under close supervision. Probably, many signs of possible adverse reactions such as confusion, weakness, loss of appetite, tremor or depression are often mistakenly attributed to old age and remain unreported. Fluoroquinolones can cause QT interval prolongation. Therefore, they should be avoided in patients with known prolongation of the QT interval, patients with uncorrected hypokalaemia or hypomagnesaemia and patients receiving class IA (e.g. quinidine, procainamide) or class III (e.g. amiodarone, sotalol) antiarrhythmic agents. Chondrotoxicity of fluoroquinolones, as observed in immature animals, has led to restricted use in paediatric patients, but there is no indication that similar effects could occur in joint cartilage of adults. Tendinitis and tendon ruptures have occurred in rare cases as late as several months after treatment with some fluoroquinolones. Chronic renal diseases, concomitant use of corticosteroids and age over 60 years have been recognised as risk factors for fluoroquinolone-induced tendon disorders. Overall, the widely used fluoroquinolones discussed in this review are generally well tolerated. Nevertheless, as with all drugs, their specific adverse effect profiles must be considered when they are chosen for treatment of bacterial infections. Because of physiological changes in renal function and in case of certain comorbidities, some special considerations are necessary when fluoroquinolones are used to treat elderly patients. PMID- 12641487 TI - Molecular therapies for viral hepatitis. AB - Current treatment modalities available for hepatitis B virus (HBV) or hepatitis C virus (HCV) infections are not efficient. The enormous disease burden caused by these two infections makes the development of novel therapies critical. For HCV, the development of an effective vaccine is urgent in view of the escalating number of infected individuals. Molecular therapies for HBV and HCV infection can be directed at reducing viral load by interfering with the life cycle of the viruses or at generating immune response against viral epitopes. The antiviral approaches consist of the delivery or expression of antisense RNAs, ribozymes or dominant negative proteins. Viral biology can be interrupted by attacking various potential targets within the two viruses. DNA-based vaccination strategies are being explored for both prevention and treatment of these diseases. Both non viral and recombinant viral vectors are being developed for safe, effective and long-term gene transfer to the liver. Although no "ideal" vector is available at this time, the ingenuity of numerous investigators is leading to the improvement of the vector systems, promising successful application of gene therapy to the prevention and treatment of viral hepatitis in the foreseeable future. PMID- 12641488 TI - Glucagon-like peptide 1 and gastric inhibitory polypeptide: potential applications in type 2 diabetes mellitus. AB - Although the insulinotropic actions of gastric inhibitory polypeptide (GIP) and glucagon-like peptide 1 (GLP-1) have been known for almost 2 decades, the incretin hormones have not yet become available for clinical application. This can be explained by their unfavourable pharmacological properties. Both hormones are rapidly inactivated by the enzyme dipeptidyl peptidase IV (DPP IV), yielding biologically inactive fragments. There have been several attempts to make use of the antidiabetogenic potential of the incretin hormones. Various analogues of GLP 1 and GIP have been generated in order to achieve resistance to DPP IV degradation. The natural GLP-1 receptor agonist exendin-4, found in the saliva of the Gila monster, has a longer biological half-life after subcutaneous injection than GLP-1, and inhibition of DPP IV using, for example, pyrrolidine derivatives provides elevated concentrations of intact, biologically active GIP and GLP-1 endogenously released from the gut. A continuous intravenous infusion of native GLP-1 for a limited time may be suitable in certain clinical situations. Numerous clinical studies are currently underway to evaluate these approaches. Therefore, an antidiabetic treatment based on incretin hormones may become available within the next 5 years. PMID- 12641486 TI - Novel drug delivery systems for insulin: clinical potential for use in the elderly. AB - Diabetes mellitus is a very common disease in the elderly and its complications are responsible for excess morbidity/mortality, loss of independence and impaired quality of life. Recent studies, while not performed in the elderly, have outlined the importance of achieving tight glycaemic control in order to prevent complications. Eighty years after its discovery, subcutaneous insulin remains a major treatment for diabetes. It is used as a first-line agent in type 1 diabetes, and in type 2 diabetes when oral antihyperglycaemic agents combined with diet and exercise fail to achieve an appropriate metabolic control. To avoid injections, other routes of insulin administration have been studied, including oral, dermal and rectal routes but they were not found to be appropriate for clinical use. Buccal or nasal insulin combined with absorption enhancers proved to have interesting properties. Inhaled insulin appears to be suitable for use in patients with diabetes because of its better bioavailability and a pharmacokinetic profile that mimics the time kinetics of insulin secretion after a meal. Clinical studies were conducted among small numbers of patients with type 1 or type 2 diabetes who had been treated with subcutaneous insulin. Inhaled insulin was given three times daily, just before meals, and was combined with a bedtime subcutaneous injection of long-acting insulin. In patients with type 1 or type 2 diabetes the metabolic control achieved with inhaled insulin was similar to that obtained with a subcutaneous insulin regimen. Tolerance of inhaled insulin was good and treatment satisfaction was better than that with the subcutaneous regimen. Insulin inhalation appears to be an interesting way of insulin delivery for elderly patients with diabetes. However, no studies have been conducted in elderly patients with diabetes to assess this route's acceptability, convenience and ease of use in this particular population. In addition, it is necessary to conduct pharmacokinetic studies in the elderly because lung aging might reduce the bioavailability of inhaled insulin. PMID- 12641489 TI - Dendritic cell vaccine design: strategies for eliciting peripheral tolerance as therapy of autoimmune diseases. AB - Dendritic cells (DC), as potent antigen-presenting cells (APC), constitute a complex system of cells that initiate and regulate immune responses that result in two opposite outcomes: immunity or tolerance. The fine regulation of these two distinct functions is not completely understood. After loading with antigen, DC exhibit the properties of both antigen and adjuvant, the functional components of vaccines. For a long time, attention has focused on the exceptional ability of DC as professional APC capable of eliciting T and B cell-mediated responses, and on their potential as immunotherapy in cancer. DC exhibit both heterogeneity and plasticity. On the one hand, distinct DC subsets exhibit distinct functions. On the other hand, DC functions can be altered by the cytokine environment or other factors. There is increasing evidence that DC could be used as a tool to induce peripheral tolerance. Because DC-based immunotherapy in autoimmune diseases depends on tolerogenic DC, discerning markers for tolerogenic DC is of great importance. Immature DC, plasmacytoid DC and interleukin-10-modified DC can mediate immune tolerance by inducing T-cell anergy or T-helper type 2 responses. Several possibilities exist for rational modulation of DC to achieve therapeutic tolerance against autoimmune diseases. The final goal is to create optimal prerequisites to use autologous DC that are prepared from the individual patient with autoimmune disease, to render such DC tolerogenic by exposure in vitro to factors that promote tolerogenicity, and to re-infuse these pretreated DC to the patient in order to treat the ongoing autoimmune disease and prevent its future exacerbation. PMID- 12641490 TI - Inhibitors of p38 mitogen-activated protein kinase: potential as anti inflammatory agents in asthma? AB - Asthma is an inflammatory disease of the airways, which in patients with mild to moderate symptoms is adequately controlled by either beta(2)-adrenoceptor agonists or corticosteroids, or a combination of both. Despite this, there are classes of patients that fail to respond to these treatments. In addition, there is a general trend towards increasing morbidity and mortality due to asthma, which suggests that there is a need for new and improved treatments. The p38 mitogen-activated protein kinases (MAPKs) represent a point of convergence for multiple signalling processes that are activated in inflammation and that impact on a diverse range of events that are important in inflammation. Small molecule pyridinyl imidazole inhibitors of p38 MAPK have proved to be highly effective in reducing various parameters of inflammation, in particular cytokine expression. Like corticosteroids, inhibitors of p38 MAPK appear to be able to repress gene expression at multiple levels, for example, by transcriptional, posttranscriptional and translational repression, and this raises the possibility of a similarly broad spectrum of anti-inflammatory activities. Indeed these molecules have proved to be effective in numerous in vitro and in vivo models of inflammation and septicaemia, which suggests that such compounds may be effective as therapeutic agents against inflammatory disorders. Despite these very promising indications of the possible therapeutic use of p38 MAPK inhibitors, a number of events that are p38-dependent are in fact also beneficial to the resolution or modulation of diseases such as asthma. We conclude that the overall effect of p38 MAPK inhibition would be beneficial in inflammatory diseases such as rheumatoid arthritis and asthma. However, these drugs may result in a complex phenotype that will require careful evaluation. Currently, a number of second or third generation inhibitors of p38 MAPK are being tested in phase I and phase II clinical trials. PMID- 12641491 TI - Prostate cancer: advances in immunotherapy. AB - The absence of curative therapies for advanced or recurrent forms of prostate cancer has prompted a vigorous search for novel treatment strategies. Immunotherapy encompasses one particularly promising systemic approach to the treatment of prostate cancer. Immune-based strategies for treating prostate cancer have recently been facilitated by the identification of a number of prostate tissue/tumour antigens that can be targeted, either by antibody or T cells, to promote prostate tumour cell injury or death. These same prostate antigens can also be used for the construction of vaccines to induce prostate specific T cell-mediated immunity. Greater insight into specific mechanisms that govern antigen-specific T cell activation has brought with it a number of innovative methods to induce and enhance T cell-mediated responses against prostate tumours. For instance, autologous dendritic cells loaded with prostate antigens have proved useful to induce prostate-specific T cell activation. Similarly, in vivo manipulations of T cell costimulatory pathway receptors can greatly facilitate tumour-specific T cell activation and potentiate T cell mediated responses against a number of malignancies, including prostate cancer. For example, blocking T cell cytotoxic lymphocyte-associated antigen 4 (CTLA-4) receptor binding to its ligand prevents the down-regulation of T cell responses and can even potentiate T cell antitumoural immunity in mouse models of prostate cancer. Androgen ablation (AA) may induce prostate tumour/tissue-specific T cell mediated inflammation and, as such, a phase II trial is currently in progress to ascertain whether CTLA-4 blockade can enhance AA-induced treatment responses in patients with advanced prostate cancer. Nevertheless, further basic and clinical investigation is still required to establish immunotherapy as a true prostate cancer treatment option. PMID- 12641492 TI - Spotlight on etanercept in rheumatoid arthritis, psoriatic arthritis and juvenile rheumatoid arthritis. AB - Etanercept (Enbrel) is a subcutaneously administered biological response modifier that binds and inactivates tumour necrosis factor-alpha, a proinflammatory cytokine. In patients with early active rheumatoid arthritis, etanercept 25mg twice weekly was associated with a more rapid improvement in disease activity and a significantly greater cumulative response than methotrexate over 12 months of treatment in a randomised, double-blind trial. In addition, etanercept recipients showed a slower rate of radiographic progression and a more rapid improvement in quality of life than methotrexate recipients. The efficacy of etanercept was maintained at 3 years' follow-up. Etanercept was also significantly better than placebo at reducing disease activity in patients who had an inadequate response to previous treatment with disease-modifying antirheumatic drugs (DMARDs) in several well controlled trials. At study end (after 3 or 6 months' treatment), the percentage of patients achieving an American College of Rheumatology 20% (ACR20) response with etanercept (25mg or 16 mg/m(2) twice weekly) was 59-75% as monotherapy and 71% in combination with methotrexate; corresponding placebo response rates were 11-14% and 27%, respectively. Response has been maintained in patients who continued treatment for up to 5 years. In patients with psoriatic arthritis, etanercept 25mg twice weekly significantly reduced disease activity and improved skin lesions in two double-blind, placebo-controlled, 12- to 24-week trials. In the 24-week study, ACR20 response rates (50 vs 13%), psoriatic arthritis response rates (70 vs 23%) and the median improvement in skin lesions (33 vs 0%) were significantly greater in etanercept than in placebo recipients. In patients with polyarticular-course juvenile rheumatoid arthritis, etanercept resulted in improvements in all measures of disease activity and was significantly more effective than placebo at reducing disease flare. Eighty percent of patients receiving etanercept achieved a > or =30% reduction in disease activity over 7 months of treatment, and this was maintained for up to 2 years in a trial extension. Etanercept was generally well tolerated in children and adults in clinical trials; the most commonly occurring adverse effects included injection site reactions, infection, headache, rhinitis and dizziness. In conclusion, etanercept has emerged as an important new treatment option in inflammatory arthritis. Etanercept provides rapid and sustained improvements in disease activity in patients with early and DMARD-refractory rheumatoid arthritis and has been shown to inhibit radiographic progression in those with early disease. Well controlled studies have also demonstrated the efficacy of etanercept in patients with psoriatic arthritis or polyarticular-course juvenile rheumatoid arthritis. PMID- 12641493 TI - Extraction of cells for single-stage seeding of vascular-bypass grafts. AB - Experimental data are reported for the seeding of prosthetic vascular grafts with either mesothelial or endothelial cells as part of a research strategy in tissue engineering with the aim of improving graft patency and developing new techniques for single-stage cell extraction and seeding that would give a step reduction in surgery time. New data are reported for two different sources of cells, peritoneal lavage and subcutaneous fat. All experiments were undertaken in patients undergoing abdominal aortic aneurysm repair. Cells extracted from peritoneal lavage were insufficient for a single-stage seeding process. Subcutaneous fat was processed using either a positive cell-extraction method using CD31 Dynabeads or by a negative extraction method using CDw90-coated magnetic beads. Only positive cell extraction gave reliably sufficient numbers of endothelial cells as a source for single-stage seeding of vascular grafts. PMID- 12641494 TI - Bioactive natural compounds for the treatment of gastrointestinal disorders. AB - Many healthy subjects and patients are taking natural bioactive products for the prevention and treatment of multiple conditions, including gastrointestinal disorders. Based on current evidence, the scientific validity of the use of many of these commercial compounds by the general public is severely limited, with quality control and regulatory issues continuing to be a concern. Nevertheless, there is sufficient preliminary data to warrant further research of these products in order to identify novel compounds for potential clinical use in addition to performing formal randomized controlled clinical trials of the commercial preparations. PMID- 12641496 TI - Review article: Pancreatic function testing. AB - Pancreatic function tests are most commonly used to diagnose chronic pancreatitis. These tests include tests which document exocrine or endocrine gland insufficiency and tests which instead measure gradations of decreased secretory capacity. The tests in the former category generally become abnormal when advanced, longstanding chronic pancreatitis is present. Tests in the latter category, however, have the potential to detect chronic pancreatitis at an earlier stage than other available diagnostic tests, including commonly used imaging tests such as computed tomography and endoscopic retrograde cholangiopancreatography. This potential advantage of diagnostic sensitivity is, however, counterweighed by the fact that these tests which measure stimulated secretory capacity are only available at a very few referral centres. This article will review the variety of pancreatic function tests and describe their rationale, accuracy, cost, and clinical usefulness. PMID- 12641495 TI - Insulin and dexamethasone induce GLUT4 gene expression in foetal brown adipocytes: synergistic effect through CCAAT/enhancer-binding protein alpha. AB - Treatment of foetal brown adipocytes in primary culture with either dexamethasone or insulin, at physiological concentrations, for 24 h up-regulates the expression of the GLUT4 gene, producing a synergistic effect on mRNA accumulation (20-fold increase), in the amount of protein in the total membrane fraction (8-fold increase) and in the transactivation of a full-promoter GLUT4 -chloramphenicol acetyltransferase gene ( CAT ) construct (7-fold increase). However, GLUT1 expression remains essentially unmodified regardless of the presence of the hormones. As a consequence, exposure of brown adipocytes to dexamethasone and insulin results in a dramatic increase of glucose uptake (12-fold). Dexamethasone induces the expression of CCAAT/enhancer-binding protein (C/EBP) alpha, insulin promotes myocyte enhancer factor-2 DNA-binding activity and both combined produces a significant increase in C/EBPalpha DNA-binding activity. Moreover, co transfection with a wild-type C/EBPalpha construct transactivates a full-promoter GLUT4 - CAT fusion gene, whereas a dominant-negative C/EBPalpha expression vector impairs the hormonal effects. Our results show that the synergism between insulin and glucocorticoids on glucose uptake is a consequence of the activation of the GLUT4 promoter by the transcription factor C/EBPalpha. PMID- 12641497 TI - Systematic review: Rabeprazole-based therapies in Helicobacter pylori eradication. AB - AIM: To perform a systematic review of the efficacy of rabeprazole-based therapies in Helicobacter pylori eradication, and to conduct a meta-analysis comparing the efficacy of rabeprazole and other proton pump inhibitors when co prescribed with antibiotics. METHODS: Studies evaluating rabeprazole plus antibiotics were considered. Only randomized trials comparing rabeprazole and other proton pump inhibitors with antibiotics, and differing only in the proton pump inhibitor, were included in the meta-analysis. Electronic and manual bibliographic searches were conducted. The percentage (weighted mean) of successful eradication was calculated. Meta-analysis was performed by combining the odds ratios (OR) of the individual studies. RESULTS: The eradication rates were as follows: 14-day rabeprazole-amoxicillin, 73%; rabeprazole-amoxicillin clarithromycin for 3, 5, 7 and 10 days, 44%, 72%, 78% and 75%, respectively; low dose rabeprazole (20 mg/day), amoxicillin and clarithromycin for 7 days, 81%; high-dose rabeprazole (40 mg/day), amoxicillin and clarithromycin for 7 days, 75%; 7-day rabeprazole-clarithromycin-nitroimidazole, 85%. Twelve comparative studies were included in the meta-analysis. The eradication rate with rabeprazole plus antibiotics was 79%; it was 77% with other proton pump inhibitors (OR = 1.15; 95% confidence interval, 0.93-1.42). Sub-analysis comparing rabeprazole at low doses (10 mg b.d.) with other proton pump inhibitors at standard doses (omeprazole 20 mg b.d. or lansoprazole 30 mg b.d.) showed no differences. CONCLUSIONS: Rabeprazole achieves similar H. pylori eradication rates to omeprazole and lansoprazole when co-prescribed with antibiotics. Low doses of rabeprazole (10 mg b.d.), when administered with two antibiotics, may be sufficient to eradicate H. pylori infection. PMID- 12641498 TI - Cost-effectiveness analysis of therapeutic strategies for patients with chronic hepatitis C previously not responding to interferon. AB - BACKGROUND: The efficacy of combination therapy in patients with chronic hepatitis C previously not responding to interferon monotherapy is lower than that in naive patients, and there has been no economic evaluation in this population. AIM: To develop a cost-effectiveness analysis of therapeutic regimens with interferon-alpha and ribavirin in previous interferon non-responders. METHODS: A Markov simulation model was used to project the clinical and economic outcomes of five different therapeutic strategies, including a 'no treatment' alternative, using the health care system perspective. The efficacy data for the different doses and durations were obtained from a previously performed meta analysis. A sensitivity analysis was performed to test the robustness of the model, analysing changes in different variables. RESULTS: Applying a 3% discount rate, the standard patient on combination therapy for 12 months showed increases of 0.80 years and 1.55 quality-adjusted life years, when compared with the 'no treatment' strategy. This option led to an incremental cost-effectiveness ratio of 11,767 euros per year of life gained and 6073 euros per quality-adjusted life year. CONCLUSIONS: Combination therapy with interferon plus ribavirin is cost effective in previous interferon non-responders and is within the range of some well-accepted medical interventions in our health care system. PMID- 12641499 TI - Eradication of Helicobacter pylori increases nocturnal intragastric acidity during dosing with rabeprazole, omeprazole, lansoprazole and placebo. AB - BACKGROUND: The eradication of Helicobacter pylori decreases the antisecretory activity of omeprazole and lansoprazole. Rabeprazole is a potent proton pump inhibitor that may not be affected as greatly by H. pylori status. AIM: To compare the effect of H. pylori eradication on intragastric acidity and plasma gastrin during dosing with lansoprazole, omeprazole, rabeprazole and placebo. METHODS: Twenty-four healthy H. pylori-infected volunteers were studied on day 7 of dosing with placebo, lansoprazole 30 mg, omeprazole 20 mg and rabeprazole 20 mg, before and at least 5 weeks after H. pylori eradication. On each occasion, the 24-h intragastric acidity was measured by gastric aspiration. Plasma gastrin concentrations were measured hourly from 08.00 to 13.00 h. RESULTS: Sixteen subjects completed the study. For all three drugs and placebo, H. pylori eradication increased intragastric acidity, particularly nocturnal acidity, and decreased plasma gastrin. There were no differences between the three drugs with respect to 24-h acidity, percentage of time pH > 4 or 5-h plasma gastrin, either before or after H. pylori eradication. Before eradication, the percentage nocturnal time at pH > 3 was significantly greater during rabeprazole than during lanso-prazole dosing. CONCLUSIONS: The increase in intragastric acidity seen after H. pylori eradication during dosing with proton pump inhibitors is a drug class effect, particularly affecting nocturnal acid control. This is related to increased spontaneous intragastric acidity after H. pylori eradication. PMID- 12641500 TI - Low-dose or standard-dose proton pump inhibitors for maintenance therapy of gastro-oesophageal reflux disease: a cost-effectiveness analysis. AB - BACKGROUND: Studies on the use of low-dose proton pump inhibitor for the maintenance therapy of gastro-oesophageal reflux disease have shown that it might be comparable with standard-dose proton pump inhibitor treatment and superior to standard-dose histamine-2 receptor antagonist therapy. AIM: To compare the impact of standard-dose histamine-2 receptor antagonist, low-dose proton pump inhibitor and standard-dose proton pump inhibitor treatment for the maintenance therapy of gastro-oesophageal reflux disease on symptom control and health care resource utilization from the perspective of a public health organization in Hong Kong. METHODS: A Markov model was designed to simulate, over 12 months, the economic and clinical outcomes of gastro-oesophageal reflux disease patients treated with standard-dose histamine-2 receptor antagonist, low-dose proton pump inhibitor and standard-dose proton pump inhibitor. The transition probabilities were derived from the literature. Resource utilization was retrieved from a group of gastro oesophageal reflux disease patients in Hong Kong. Sensitivity analysis was conducted to examine the robustness of the model. RESULTS: The standard-dose proton pump inhibitor strategy was associated with the highest numbers of symptom free patient-years (0.954 years) and quality-adjusted life-years gained (0.999 years), followed by low-dose proton pump inhibitor and standard-dose histamine-2 receptor antagonist. The direct medical cost per patient in the standard-dose proton pump inhibitor group (904 US dollars) was lower than those of the low-dose proton pump inhibitor and standard-dose histamine-2 receptor antagonist groups. CONCLUSIONS: The standard-dose proton pump inhibitor strategy appears to be the most effective and least costly for the maintenance management of patients with gastro-oesophageal reflux disease in Hong Kong. PMID- 12641501 TI - A rapid, low-dose, 13C-urea tablet for the detection of Helicobacter pylori infection before and after treatment. AB - BACKGROUND: A new urea breath test (UBT) has been described which uses a tablet formulation of 13C-urea with citric acid and allows breath sampling to be performed as early as 10 min after ingestion of the tablet. AIM: To assess the diagnostic accuracy of tablet-based 13C-UBTs (50 and 100 mg 13C-urea) before and after Helicobacter pylori eradication treatment, compared with an endoscopy gold standard and a conventional 13C-UBT (75 mg 13C-urea). METHODS: Two hundred dyspeptic patients underwent endoscopy, followed by tablet-based 13C-UBTs (50 and 100 mg 13C-urea) and a conventional 13C-UBT (75 mg 13C-urea). H. pylori-infected patients were prescribed treatment and asked to return 4-6 weeks after the end of therapy for repeat endoscopy and 13C-UBTs. RESULTS: One hundred and thirteen patients were infected with H. pylori. The sensitivity and specificity of the conventional 13C-UBT were both 100%; the sensitivity and specificity of the 100 mg tablet-based 13C-UBT were 100% and 98.85%, respectively. For the 50-mg tablet based 13C-UBT, cut-off values of the difference over baseline of between 1.65 and 3.15 provided a sensitivity and specificity of 100%. At follow-up, the sensitivity and specificity of the conventional and 100-mg tablet-based 13C-UBTs were both 100%. For the 50-mg tablet-based 13C-UBT, cut-off values of the difference over baseline of between 1.49 and 1.56 gave a sensitivity and specificity of 100%. CONCLUSIONS: New 10-min 13C-UBTs using tablet formulations of 13C-urea with citric acid are reliable for the assessment of H. pylori status pre- and post-treatment. PMID- 12641502 TI - Effect of treatment of Helicobacter pylori on the prevention of gastroduodenal ulcers in patients receiving long-term NSAIDs: a double-blind, placebo-controlled trial. AB - BACKGROUND: There is controversy as to whether Helicobacter pylori and non steroidal anti-inflammatory drugs interact to cause peptic ulcers. AIM: To study whether the eradication of H. pylori in patients on long-term non-steroidal anti inflammatory drug therapy prevents the development of ulcers. METHODS: Patients infected with H. pylori whilst receiving long-term non-steroidal anti inflammatory drug therapy, but with no ulcers at baseline endoscopy, were randomized to receive either triple antibiotic therapy (metronidazole 300 mg, clarithromycin 250 mg and amoxicillin 500 mg, given four times daily; n = 70) or placebo (n = 70) for 2 weeks. Non-steroidal anti-inflammatory drugs were continued throughout the study period. Endoscopy was repeated 12 weeks after the end of treatment. The development of ulcers was compared between the two groups. RESULTS: Endoscopy at 12 weeks revealed peptic ulcer development in five [7%; 95% confidence interval (CI), 2-16] of the patients who received triple therapy and in six (9%; 95% CI, 3-18) of those who received placebo (P = 1.00). No significant difference in the development of ulcers was found between patients with persistent H. pylori infection (7/80; 9%; 95% CI, 4-17) and those with the eradication of H. pylori (4/52; 8%; 95% CI, 2-19) (P = 1.00). CONCLUSIONS: The eradication of H. pylori in patients receiving long-term treatment with non steroidal anti-inflammatory drugs did not prevent ulcer development. However, because the rate of ulcer development was low, a study with a larger sample size is required to confirm this finding. PMID- 12641503 TI - Therapy of osteoporosis in patients with Crohn's disease: a randomized study comparing sodium fluoride and ibandronate. AB - BACKGROUND: Osteoporosis is a frequent complication in Crohn's disease. Although the efficacy of both sodium fluoride and aminobisphosphonates in postmenopausal osteoporosis has been investigated in long-term therapy studies, no long-term results are available regarding the effect of these agents in the management of osteoporosis in patients with Crohn's disease. METHODS: Eighty-four patients with Crohn's disease and pathological bone mineral density findings were randomized to receive either vitamin D3 (1000 IU) and calcium citrate (800 mg) daily (group A) or sodium fluoride (25 mg b.d., group B) or intravenous ibandronate (1 mg every 3 months, group C) in addition to daily calcium/vitamin D substitution. On admission to the study and after 12 and 27 months, patients underwent dual-energy X-ray absorptiometry and radiological examination of the spine. RESULTS: Sixty eight patients completed the 1-year observation period and were available for the intention-to-treat analysis. No new vertebral fractures were diagnosed. In group A, lumbar bone density increased by 2.6% (P = 0.066, N.S.), in group B by 5.7% (P = 0.003) and in group C by 5.4% (P = 0.003). Therapy with sodium fluoride was associated with an increase in osteocalcin (N.S.), whereas administration of ibandronate was associated with a decrease in the resorption parameter, carboxy terminal cross-linked type-I collagen telopeptide (P < 0.05). Both sodium fluoride and ibandronate resulted in significant decreases in the serum concentration of osteoprotegerin after 9 months (P < 0.001). CONCLUSIONS: The findings of the present study show that both sodium fluoride and ibandronate are effective in combination with calcium and vitamin D substitution in the management of osteopenia and osteoporosis in patients with Crohn's disease. Both agents are safe and well tolerated, and induce continuous increases in lumbar bone density. PMID- 12641504 TI - Non-steroidal anti-inflammatory drugs, aspirin and newly diagnosed colitis: a case-control study. AB - BACKGROUND: There have been a number of reports of colitis following exposure to non-steroidal anti-inflammatory drugs (NSAIDs) and salicylates. AIM: To conduct a case-control analysis of new cases of colitis, with particular reference to the usage of NSAIDs and salicylates prior to the development of the disease. METHODS: One hundred and five consecutive new cases of colitis presenting to a single gastroenterologist were questioned about their recent usage of NSAIDs and salicylates. For comparison, the frequency of usage of these compounds was studied in two groups of 105 age- and sex-matched controls taken from hospital in patients and community cases attending the Accident and Emergency Department. RESULTS: Of the 105 cases of colitis studied, 78 patients (74%) had been taking NSAIDs or salicylates prior to or during the development of their disease. By comparison, 20% of community controls were using NSAIDs or salicylates (P < 0.001) and 30% of hospital in-patients were taking these compounds (P < 0.001). Comparison of these frequencies with those of the colitis group gave odds ratios of 9.1 (4.5, 21.9) with the community controls and 6.2 (3.2, 13.5) with the hospital controls. CONCLUSIONS: In new patients presenting with colitis, there is a significantly high frequency of antecedent exposure to NSAIDs or salicylates, supporting the concept that these agents may be important in the pathogenesis of colitis. PMID- 12641505 TI - Azathioprine, mercaptopurine and birth outcome: a population-based cohort study. AB - BACKGROUND: Data on the safety of azathioprine and mercaptopurine during pregnancy are very sparse. AIM: To examine the risk of adverse birth outcomes in women who took up prescriptions for azathioprine or mercaptopurine during pregnancy. METHODS: This is a Danish cohort study based on data from a population based prescription registry, the Danish Birth Registry and the Hospital Discharge Registry. To examine the risk of congenital malformations, we included nine pregnancies exposed 30 days before conception or during the first trimester. To examine perinatal mortality, pre-term birth and low birth weight, we included 10 pregnancies exposed during the entire pregnancy. Eleven different exposed women were included in the study. Outcomes were compared with those of 19 418 pregnancies in which no drugs were prescribed to the mothers. RESULTS: Fifty-five per cent of the exposed women had inflammatory bowel disease and 45% other diseases. Adjusted odds ratios for congenital malformations, perinatal mortality, pre-term birth and low birth weight were 6.7 (95% confidence interval, 1.4-32.4), 20.0 (2.5-161.4), 6.6 (1.7-25.9) and 3.8 (0.4-33.3), respectively. CONCLUSIONS: Our results suggest that there is an increased risk of congenital malformations, perinatal mortality and pre-term birth in children born to women treated with azathioprine or mercaptopurine during pregnancy. More data are needed to determine whether the associations are causal or occur through confounding. PMID- 12641506 TI - Health-related anxiety and the effect of open-access endoscopy in US patients with dyspepsia. AB - BACKGROUND: The role of endoscopy in dyspepsia is the subject of debate. The detection of lesions is infrequent, but patients may benefit from the knowledge that the examination is normal. We sought to determine the prevalence of health related anxiety in dyspeptic patients referred for open-access endoscopy and to investigate the effect of endoscopy on health-related anxiety. METHODS: Consecutive patients referred for open-access endoscopy from primary care were studied using a validated questionnaire for health-related anxiety before and after endoscopy, at 1 month and 6 months. Symptoms were assessed using a validated questionnaire at 1 and 6 months. RESULTS: One hundred and nine patients were studied (69 women and 40 men; mean age, 49 +/- 15 years). Thirty-six of the 109 patients (33%) had high anxiety scores at baseline (mean score, 41 +/- 1), which decreased after endoscopy to 35 +/- 1 (P < 0.05). The changes persisted at 1 month (33 +/- 1) and 6 months (33 +/- 1). Endoscopic findings were as follows: normal examination, 120; erosive oesophagitis, 11 (Grade A); erosive duodenitis (all Helicobacter pylori-negative), 6. Scales for preoccupation with health and fear of illness and death showed significant improvement after endoscopy, and the effects were preserved for 6 months. Anxiety scores in our population were substantially higher than in a corresponding UK population. CONCLUSIONS: Health related anxiety is common in dyspeptic patients referred for endoscopy. Endoscopy decreases the preoccupation with health and fear of illness and death in patients with severe anxiety, and the effects persist for 6 months. PMID- 12641507 TI - Predictors of conventional and alternative health care seeking for irritable bowel syndrome and functional dyspepsia. AB - BACKGROUND: The drivers of conventional and, especially, alternative health care use for irritable bowel syndrome and functional dyspepsia are not clear. AIM: To determine the predictors of conventional and alternative health care use for irritable bowel syndrome and functional dyspepsia. METHODS: Two hundred and seven subjects with irritable bowel syndrome or functional dyspepsia, identified from a previous population survey, were included in the study. Individuals with irritable bowel syndrome/functional dyspepsia were defined as consulters (n = 103) if they had visited their doctor for gastrointestinal symptoms more than once in the past year. Controls (n = 100) did not report having any abdominal pain. Subjects were given structured interviews to assess the Diagnostic and Statistical Manual - version IV (DSM-IV) and International Classification of Disorders - version 10 (ICD-10) psychiatric diagnosis for anxiety, depression, somatization or any psychiatric diagnosis, aspects of health care use and symptom factors. RESULTS: About one-half (n = 103, 49.8%) of community subjects with irritable bowel syndrome/functional dyspepsia had sought conventional care for gastrointestinal symptoms in the past 12 months. Lifetime rates for alternative health care use for gastrointestinal symptoms were 20.8% (n = 43). Independent predictors of conventional health care use were more frequent abdominal pain, greater interference of gastrointestinal symptoms with work and activities and a greater satisfaction with the physician-patient relationship. Being female independently predicted alternative health care use. CONCLUSIONS: Psychological morbidity did not predict conventional or alternative health care use for gastrointestinal symptoms. Other factors were more important. PMID- 12641508 TI - Longevity of silicone and polyurethane catheters in long-term enteral feeding via percutaneous endoscopic gastrostomy. AB - BACKGROUND: As percutaneous endoscopic gastrostomy (PEG) is often used for many months or years, the longevity of the feeding tubes plays an important role in the global outcome and costs of PEG. AIM: A retrospective study to evaluate the longevity of silicone and polyurethane PEG catheters. METHODS: The records of 297 patients who were fed via PEG for over 90 days were evaluated. The material of the PEG catheter, duration of follow-up, local complications, need to remove PEG because of tube deterioration or local complications and time from PEG placement to PEG removal were recorded and compared. RESULTS: Two hundred and twenty-eight patients had polyurethane and 69 had silicone PEG catheters. The follow-up ranged from 116 to 3207 days for the polyurethane group and from 98 to 1861 days for the silicone group. No differences were observed in either local complications or PEG removal because of local complications. Tube deterioration causing PEG removal occurred in 36 of the 228 polyurethane PEG catheters and in 25 of the 69 silicone PEG catheters (P = 0.0005). Tube deterioration occurred significantly earlier in the 25 silicone catheters than in the 36 polyurethane catheters. The mean time from PEG placement to PEG removal was 287 days (95% confidence interval, 239-335) for silicone tubes and 573.9 days (95% confidence interval, 425-723) for polyurethane tubes (P = 0.0024). CONCLUSION: Polyurethane PEG catheters seem to be more resistant to deterioration than silicone PEG catheters, and at present they should be preferred for long-term enteral feeding via PEG. PMID- 12641509 TI - Review article: indicators and predictors of response to anti-viral therapy in chronic hepatitis C. AB - The complications of chronic hepatitis C, including cirrhosis and hepatocellular carcinoma, are expected to increase dramatically world-wide over the next 10-20 years. Immunomodulatory/anti-viral therapy, employing interferon alfa both alone and in combination with ribavirin, affords the only effective treatment for hepatitis C. Accurate early prediction of response to interferon therapy may decrease or eliminate unnecessary or ineffective treatment, permit greater flexibility in tailoring therapy on an individual basis, and enhance the cost effectiveness of treatment. Liver biopsy provides valuable information about the baseline severity and subsequent progression of hepatitis C. Severe fibrosis or cirrhosis on the pre-treatment liver biopsy is associated with decreased response rates. The measurement of viral RNA levels and genotyping may be used to optimize individual patient treatment. Genotype non-1 and a low viral load are the most significant pre-treatment indicators of sustained virological response. The most reliable predictor of a poor virological response is continued seropositivity for viral RNA during therapy. Therefore, a decision to stop or continue treatment can be based on a positive viral RNA test at 12 weeks for interferon-naive patients receiving interferon or pegylated interferon therapy. PMID- 12641510 TI - Review article: visceral hypersensitivity. AB - Visceral hypersensitivity is highly prevalent in all functional bowel disorders. Most also demonstrate wider patterns of somatic referral of intestinal pain or discomfort. This hypersensitivity may explain the symptoms as the sensitive gut can be more easily provoked by normal or abnormal motor events in the gut. Visceral hypersensitivity may increase during psychosocial stress and during periods of symptom exacerbation, although this requires confirmation. Pharmacological therapy to reduce visceral hypersensitivity is now possible using antagonists to neurotransmitters, opening up an exciting new era for the treatment of functional gastrointestinal disorders. PMID- 12641511 TI - Gut-focused hypnotherapy normalizes disordered rectal sensitivity in patients with irritable bowel syndrome. AB - BACKGROUND: We have previously shown that hypnotherapy alters rectal sensitivity in some patients with irritable bowel syndrome. However, this previous study used incremental volume distension of a latex balloon, which might be susceptible to subject response bias and might compromise the assessment of compliance. In addition, the study group was symptomatically rather than physiologically defined. AIM: To assess the effect of hypnotherapy on rectal sensitivity in hypersensitive, hyposensitive and normally sensitive irritable bowel syndrome patients using a distension technique (barostat) that addresses these technical issues. METHODS: Twenty-three irritable bowel syndrome (Rome I) patients (aged 24 72 years) were assessed before and after 12 weeks of hypnotherapy in terms of rectal sensitivity, symptomatology, anxiety and depression. Normal values for sensitivity were established in 17 healthy volunteers (aged 20-55 years). RESULTS: Compared with controls, 10 patients were hypersensitive, seven hyposensitive and six normally sensitive before treatment. Following hypnotherapy, the mean pain sensory threshold increased in the hypersensitive group (P = 0.04) and decreased in the hyposensitive group, although the latter failed to reach statistical significance (P = 0.19). Normal sensory perception was unchanged. Sensory improvement in the hypersensitive patients tended to correlate with a reduction in abdominal pain (r = 0.714, P = 0.07). CONCLUSION: Hypnotherapy improves abnormal sensory perception in irritable bowel syndrome, leaving normal sensation unchanged. PMID- 12641512 TI - The prevalence, patterns and impact of irritable bowel syndrome: an international survey of 40,000 subjects. AB - AIM: To determine the prevalence, symptom pattern and impact of the irritable bowel syndrome, across eight European countries, using a standardized methodology. METHODS: A community survey of 41 984 individuals was performed using quota sampling and random digit telephone dialing to identify those with diagnosed irritable bowel syndrome or those meeting diagnostic criteria, followed by in-depth interviews. RESULTS: The overall prevalence was 11.5% (6.2-12%); 9.6% had current symptoms, 4.8% had been formally diagnosed and a further 2.9%, 4.2% and 6.5% met the Rome II, Rome I or Manning criteria, respectively. Bowel habit classification varied by criteria: 63% had an 'alternating' bowel habit by Rome II vs. 21% by self-report. On average, 69% reported symptoms lasting for 1 h, twice daily, for 7 days a month. Irritable bowel syndrome sufferers reported more peptic ulcer (13% vs. 6%), reflux (21% vs. 7%) and appendectomy (17% vs. 11%), but not hysterectomy, cholecystectomy or bladder procedures. Ninety per cent had consulted in primary care and 17% in hospital; 69% had used medication. Irritable bowel syndrome substantially interfered with lifestyle and caused absenteeism. CONCLUSIONS: Irritable bowel syndrome is common with major effects on lifestyle and health care. The majority of cases are undiagnosed and the prevalence varies strikingly between countries. Diagnostic criteria are associated with varying prevalences and bowel habit sub-types. This limits their utility in clinical practice and the transferability of research findings using them. PMID- 12641513 TI - General practitioner and hospital specialist attitudes to functional gastrointestinal disorders. AB - BACKGROUND: Functional gastrointestinal symptoms generate a large workload in primary care. Research on functional gastrointestinal disorders is focused on hospital patients, but these patients may differ from those managed in primary care. AIM: To investigate any differences in attitudes of general practitioners and hospital specialists towards functional gastrointestinal illnesses. METHODS: A questionnaire was sent to 200 general practitioners and 200 British Society of Gastroenterology members. RESULTS: The response rate was 76%. Sixty-two general practitioners believed that functional gastrointestinal symptoms represented a 'real' currently unexplained gastrointestinal disorder, and 67 believed such symptoms probably represented somatization of a psychological illness. In contrast, most consultants (120) believed that functional gastrointestinal symptoms represented a 'real' gastrointestinal disorder, with only 36 perceiving them to have a psychological basis (chi2 = 26.7, P < 0.001). More consultants than general practitioners believed that the understanding of functional gastrointestinal disorders had improved in the last 20 years (chi2 = 4.31, P < 0.05). Most consultants and most general practitioners thought that treatment for these disorders had not improved over this period. Only 21% of general practitioners had heard of the Manning criteria for the diagnosis of irritable bowel syndrome, compared to 81% of consultants (chi2 = 107, P < 0.0001); 12% of general practitioners and 83% of consultants had heard of the Rome criteria for the diagnosis of functional gastrointestinal disorders (chi2 = 154, P < 0.0001); 37% of consultants used the Manning criteria and 40% used the Rome criteria; 11% of general practitioners used the Manning criteria and 3% used the Rome criteria. CONCLUSIONS: General practitioners and consultants have differing views on functional gastrointestinal disorders. In both primary and secondary care, most doctors do not use diagnostic criteria. Further research on the factors used to diagnose functional gastrointestinal disorders in primary care is warranted. PMID- 12641514 TI - Trends in oesophageal and gastric cancer incidence, mortality and survival in England and Wales 1971-1998/1999. AB - BACKGROUND: In England and Wales, 7% of cancers diagnosed in 1997 were oesophageal or gastric cancer. AIM: To review the epidemiology of these cancers in England and Wales using cancer registration data from 1971 to 1998 and mortality data from 1971 to 1999. METHODS: We calculated age-specific and age standardised incidence and mortality rates; crude and relative survival by age group; age-standardised relative survival; and relative survival by socio economic status. RESULTS: The age-standardised incidence of oesophageal cancer increased from 7.6 to 12.8 per 100,000 men and from 4.2 to 5.7 per 100,000 women. The incidence of gastric cancer decreased from 31.8 to 18.9 per 100,000 men and from 15.1 to 7.3 per 100,000 women. For both cancers, the 1- and 5-year age standardised relative survival improved for men and women from 1971-1975 to 1992 1994; the 5-year age-standardised relative survival was significantly higher in women than in men diagnosed in 1992-1994; for some periods from 1971-1975 to 1986 1990, the 1-year relative survival was significantly higher in the least deprived men than in the most deprived men. CONCLUSIONS: Survival improved for both cancers over the study period, but there were unexplained differences related to sex and deprivation. PMID- 12641515 TI - Trends in the subsite and morphology of oesophageal and gastric cancer in England and Wales 1971-1998. AB - BACKGROUND: The incidence of adenocarcinoma of the oesophagus and gastric cardia has increased in many countries. AIM: To describe the trends in the subsite and morphology of oesophageal and gastric cancer using cancer registry data from 1971 to 1998. METHODS: We calculated the overall age-standardised incidence in each year; the age-standardised incidence by subsite, by morphology and by subsite and morphology; and the ratio of the rates in men and women in 1971 and 1998. RESULTS: The incidence of oesophageal adenocarcinoma increased from 1.5 to 7.0 per 100,000 men and from 0.4 to 1.5 per 100,000 women. The incidence of cancer at the cardia also increased, from 2.0 to 5.4 per 100,000 men and from 0.6 to 1.4 per 100,000 women, but the incidence of gastric cancer without a specified subsite decreased markedly from 21.3 to 9.3 per 100,000 men and from 10.7 to 4.2 per 100,000 women. CONCLUSIONS: Although some of the increase in the incidence of adenocarcinoma of the gastric cardia is probably real, this interpretation is limited by the proportion of cancers without specified subsites or morphologies. PMID- 12641516 TI - Oesophageal cancer and cachexia: the effect of short-term treatment with thalidomide on weight loss and lean body mass. AB - BACKGROUND: Cachexia is common in patients with advanced cancer and has a direct impact on well-being and mortality. AIM: To test the hypothesis that thalidomide can promote weight gain and lean body mass in patients with advanced oesophageal cancer. METHODS: In an open-label study, 11 patients with non-obstructing and inoperable oesophageal cancer were established on an isocaloric diet for 2 weeks, followed by 2 weeks on thalidomide, 200 mg daily. The primary end-points were weight change and lean body mass. Secondary end-points were quality of life and changes in resting energy expenditure. RESULTS: Ten patients completed the study protocol. The average caloric intake remained the same throughout the study period in all patients. Nine of 10 patients (95% confidence interval, 0.60, 0.98) lost weight on diet alone. The mean weight gain on thalidomide in the following 2 weeks was 1.29 kg (median, 1.25 kg). A similar trend was shown in the lean body mass. Eight of nine patients (95% confidence interval, 0.57, 0.98) initially lost lean body mass on diet alone (missing data in one patient). The mean gain in lean body mass on thalidomide in the following 2 weeks was 1.75 kg (median, 1.33 kg). CONCLUSIONS: Thalidomide treatment appeared to reverse the loss of weight and lean body mass over the 2-week trial period. PMID- 12641517 TI - The prophylactic use of a proton pump inhibitor before food and alcohol. AB - BACKGROUND: Patients report that the prophylactic consumption of a proton pump inhibitor minimizes gastrointestinal symptoms expected to be provoked by late night food and alcohol consumption. The efficacy of this practice has not been studied formally. AIM: To perform a randomized, double-blind, placebo-controlled trial of a single dose of lansoprazole (30 mg) taken prior to a large meal and alcohol consumption. METHODS: Study subjects were recruited randomly from local primary care and hospital physicians. Each participant (n = 56; 37 male, 19 female; mean age, 38 years) completed questionnaires before and after the meal. Approximately 90 min prior to the provocative meal, participants were witnessed taking either placebo or 30 mg lansoprazole. Bar tokens were dispensed to permit the accurate quantification of alcohol consumption (mean, 15 units). RESULTS: Forty per cent of subjects reported significant reflux symptoms. For the entire group, there was no significant difference between lansoprazole and placebo. Post prandial reflux was more frequent in those consuming > 15 units of alcohol (13/26, 50%) compared with those consuming < 15 units (7/30, 24%; P < 0.05). In the group who consumed > 15 units of alcohol, lansoprazole was associated with a lower rate of heartburn (5/15, 33%) compared with placebo (8/11, 73%; P < 0.05). CONCLUSION: A single dose of a proton pump inhibitor prior to indulgence was only associated with reduced heartburn in those consuming > 15 units of alcohol. PMID- 12641518 TI - A cost-effectiveness analysis of peginterferon alfa-2b plus ribavirin for the treatment of naive patients with chronic hepatitis C. AB - AIM: To estimate the cost-effectiveness of therapy and analyse the effect of therapy compliance in naive patients with chronic hepatitis C. METHODS: A decision analysis using the Markov model was performed for four different therapeutic strategies using peginterferon alfa-2b plus ribavirin or interferon alfa-2b plus ribavirin. Clinical data were obtained from available published reports and from the Spanish health system perspective. RESULTS: The incremental cost-effectiveness ratio of peginterferon alfa-2b plus ribavirin at a fixed dose, compared with interferon alfa-2b plus ribavirin, was 8478 euros per life year saved and 3737 euros per quality-adjusted life year gained. Good therapeutic compliance and weight-adjusted doses of ribavirin decreased the incremental cost effectiveness ratio to 1636 euros per life year saved and 721 euros per quality adjusted life year gained. In compliant genotype 1 patients, the incremental cost effectiveness ratio decreased to 916 euros per life year saved and 404 euros per quality-adjusted life year gained, with an increase from 64 to 69 years in the threshold age at which therapy was cost-effective. The sensitivity analysis demonstrated that changes in the values of the most relevant parameters do not modify the study outcomes. CONCLUSION: From the clinical and pharmaco-economics perspective, the use of decision therapeutic analysis models suggests that the most effective therapy for chronic hepatitis C is peginterferon alfa-2b plus ribavirin adjusted to patient body weight and with good compliance, particularly in genotyped patients. PMID- 12641519 TI - Role of immunosuppression in the development of quinolone-resistant Escherichia coli spontaneous bacterial peritonitis and in the mortality of E. coli spontaneous bacterial peritonitis. AB - BACKGROUND: Norfloxacin decreases the incidence of spontaneous bacterial peritonitis in cirrhotics, but promotes the appearance of quinolone-resistant Escherichia coli. AIM: : To define the characteristics of quinolone-resistant E. coli spontaneous bacterial peritonitis. METHODS: E. coli-positive ascitic fluid cultures were identified during a 6-year period. Data on quinolone-sensitive and quinolone-resistant E. coli spontaneous bacterial peritonitis were compared. RESULTS: One hundred and two E. coli-positive ascitic fluid cultures were detected. Cirrhotics accounted for 67 cases. Spontaneous bacterial peritonitis was found in 47 of the 67 (70%) cases [35 (74%) caused by quinolone-sensitive and 12 (26%) caused by quinolone-resistant E. coli]. Norfloxacin prophylaxis was higher in the quinolone-resistant group (92% vs. 6%, P < 0.001). Compared with patients with quinolone-sensitive E. coli spontaneous bacterial peritonitis, those with quinolone-resistant E. coli spontaneous bacterial peritonitis showed a higher prevalence of associated immunosuppressive factors (immunosuppressive drugs, human immunodeficiency virus infection or cancer) (92% vs. 20%, P < 0.001). Steroid therapy was independently associated with quinolone-resistant E. coli spontaneous bacterial peritonitis (odds ratio, 49; 95% confidence interval, 3.4-699; P = 0.004). The Child-Pugh score (P = 0.03), immunosuppression (P = 0.02) and renal failure (P = 0.01) were independent predictors of E. coli spontaneous bacterial peritonitis-related mortality. CONCLUSIONS: Associated immunosuppression is an important co-factor for the development of quinolone resistant E. coli spontaneous bacterial peritonitis and for E. coli spontaneous bacterial peritonitis-related mortality. PMID- 12641520 TI - Response, relapse and mucosal immune regulation after infliximab treatment in fistulating Crohn's disease. AB - BACKGROUND: Infliximab reduces mucosal inflammation in some, but not all, patients with Crohn's disease. AIM: To monitor clinical data and changes in mucosal cytokine levels after infliximab treatment to identify differences between responders and non-responders. METHODS: Twenty-six patients with fistulating Crohn's disease received three infliximab infusions at weeks 0, 2 and 6. Follow-up was for 1 year and included clinical examination, colonoscopy, ano rectal ultrasound and magnetic resonance imaging. Biopsies were taken at weeks 0, 8, 26 and 52. Cell cultures were established and analysed for tumour necrosis factor-alpha, interferon-gamma and interleukin-10 levels, and related to clinical status and fistula healing. RESULTS: Eleven of 15 patients (73%) with active disease (Crohn's disease activity index > 150) obtained remission (Crohn's disease activity index < 150) at 8 weeks. In in vitro cell cultures, there was reduced tumour necrosis factor-alpha and interleukin-10 production at week 26, with the latter persistent throughout the study period. When the disease deteriorated or relapsed, there was increased interferon-gamma production in in vitro cell cultures. Fistula healing was associated with reduced production of interferon-gamma, tumour necrosis factor-alpha and interleukin-10. CONCLUSIONS: Infliximab down-regulates mucosal immune activation in Crohn's disease. Monitoring of mucosal cytokine levels after infliximab treatment by whole biopsy cultures may be useful as interleukin-10, tumour necrosis factor-alpha and interferon-gamma production are different in responders and at relapse. PMID- 12641521 TI - Quality of life in Chinese patients with inflammatory bowel disease: validation of the Chinese translation of the Inflammatory Bowel Disease Questionnaire. AB - BACKGROUND: Health-related quality of life is an important outcome measure in inflammatory bowel disease. The Inflammatory Bowel Disease Questionnaire is a quality of life questionnaire that has not been validated previously in Chinese patients with inflammatory bowel disease. AIM: To develop and validate a Chinese translation of the Inflammatory Bowel Disease Questionnaire, specifically determining its construct validity, discriminant ability, reliability and sensitivity to change. METHODS: We developed a Chinese version of the Inflammatory Bowel Disease Questionnaire. Chinese patients with Crohn's disease and ulcerative colitis completed the Chinese Inflammatory Bowel Disease Questionnaire and visual analogue scales measuring systemic, social, bowel and emotional well-being. Patients also completed a validated Chinese SF-36 generic quality of life questionnaire, the Crohn's disease activity index or the clinical activity index for ulcerative colitis. RESULTS: One hundred and thirty-five patients (59 with Crohn's disease and 76 with ulcerative colitis) were enrolled, 99 of whom also completed the Chinese Inflammatory Bowel Disease Questionnaire for a second time. The Chinese Inflammatory Bowel Disease Questionnaire correlated well with the SF-36 for all four domains (Spearman: r = 0.55-0.80), the Crohn's disease activity index (r = -0.62-0.72) and the clinical activity index for ulcerative colitis (r = -0.44-0.68), as well as with the visual analogue scales. The Chinese Inflammatory Bowel Disease Questionnaire accurately distinguished between active and inactive disease. Test-re-test reliability showed excellent intra-class correlation (0.76-0.92; all P < 0.001). The Chinese Inflammatory Bowel Disease Questionnaire was also sensitive to changes in disease activity (P < 0.05). CONCLUSION: The Chinese Inflammatory Bowel Disease Questionnaire is a valid and reliable test that correlates well with the patients' subjective well-being and clinical disease activity. PMID- 12641522 TI - High eradication rates of Helicobacter pylori with a new sequential treatment. AB - BACKGROUND: Eradication rates of Helicobacter pylori with standard triple therapy are disappointing, and studies from several countries confirm this poor performance. AIM: To assess the eradication rate of a new sequential treatment regimen compared with conventional triple therapy for the eradication of H. pylori infection. METHODS: One thousand and forty-nine dyspeptic patients were studied prospectively. H. pylori-infected patients were randomized to receive 10 day sequential therapy [rabeprazole (40 mg daily) plus amoxicillin (1 g twice daily) for the first 5 days, followed by rabeprazole (20 mg), clarithromycin (500 mg) and tinidazole (500 mg) twice daily for the remaining 5 days] or standard 7 day therapy [corrected] [rabeprazole (20 mg), clarithromycin (500 mg) and amoxicillin (1 g) twice daily]. H. pylori status was assessed by histology, rapid urease test and 13C-urea breath test at baseline and 6 weeks or more after completion of treatment. RESULTS: Higher eradication rates were found with the sequential regimen compared to the standard regimen (intention-to-treat: 92% vs. 74%, P < 0.0001; per protocol: 95% vs. 77%, P < 0.0001). Higher eradication rates were also seen in patients with peptic ulcer disease and non-ulcer dyspepsia. In both treatments, compliance was similar (> 90%), as was the rate of side-effects, which were mild. CONCLUSIONS: This 10-day sequential treatment regimen achieves high eradication rates in peptic ulcer disease and non-ulcer dyspepsia. PMID- 12641523 TI - Stool antigen for the diagnosis of Helicobacter pylori infection in cirrhosis: comparative usefulness of three different methods. AB - BACKGROUND: Helicobacter pylori infection may lead to peptic ulcer disease, and causes significant morbidity in patients with cirrhosis. The measurement of H. pylori antigens in human stools has been proposed as a valuable, non-invasive, diagnostic tool. A number of tests have recently been commercialized. However, very few data are available on their reliability in patients with cirrhosis. AIM: To evaluate the usefulness of three new tests--HpSA (Meridian Diagnostics Inc., Cincinnati, OH, USA), Simple H. pyl (OPERON S.A., Zaragoza, Spain) and FemtoLab H. pylori (Connex, Martinsried, Germany)--in the diagnosis of H. pylori infection in cirrhotic patients. METHODS: H. pylori infection was determined in 79 cirrhotic patients (48 men, 31 women; age range, 29-82 years; mean, 62 +/- 11 years) by concordance of histology and urea breath test. The sensitivity, specificity and positive and negative predictive values of each stool test were calculated. RESULTS: According to the reference method, the sensitivities of HpSA, Simple H. pyl and FemtoLab H. pylori immunoassays were 76%, 87% and 78%, respectively, and their specificities were 93%, 62% and 79%, respectively. CONCLUSIONS: Faecal tests are non-invasive and easy-to-perform tools for the diagnosis of H. pylori infection. However, their sensitivity and specificity seem to be non-optimal in patients with cirrhosis. PMID- 12641525 TI - Research governance: implications for health library and information professionals. AB - The Research Governance Framework for Health and Social Care published by the Department of Health in 2001 provides a model of best practice and a framework for research in the health and social care sector. This article reviews the Department of Health Research Governance Framework, discusses the implications of research governance for library and information professionals undertaking research in the health- and social-care sector and recommends strategies for best practice within the information profession relating to research governance. The scope of the Framework document that covers both clinical and non-clinical research is outlined. Any research involving, amongst other issues, patients, NHS staff and use or access to NHS premises may require ethics committee approval. Particular reference is made to the roles, responsibilities and professional conduct and the systems needed to support effective research practice. Issues such as these combine to encourage the development of a quality research culture which supports best practice. Questions arise regarding the training and experience of researchers, and access to the necessary information and support. The use of the Framework to guide research practice complements the quality issues within the evidence-based practice movement and supports the ongoing development of a quality research culture. Recommendations are given in relation to the document's five domains of ethics, science, information, health and safety and finance and intellectual property. Practical recommendations are offered for incorporating research governance into research practice in ways which conform to the Framework's standards and which are particularly relevant for research practitioners in information science. Concluding comments support the use of the Research Governance Framework as a model for best practice. PMID- 12641526 TI - Using SERVQUAL in health libraries across Somerset, Devon and Cornwall. AB - This study provides the results of a survey conducted in the autumn of 2001 by ten NHS library services across Somerset, Devon and Cornwall. The aim of the project was to measure the service quality of each individual library and to provide an overall picture of the quality of library services within the south west peninsula. The survey was based on SERVQUAL, a diagnostic tool developed in the 1980s, which measures service quality in terms of customer expectations and perceptions of service. The survey results have provided the librarians with a wealth of information about service quality. The service as a whole is perceived to be not only meeting but also exceeding expectations in terms of reliability, responsiveness, empathy and assurance. For the first time, the ten health library services can measure their own service quality as well as benchmark themselves against others. PMID- 12641527 TI - Patterns and costs of printed and online journal usage. AB - This study in an academic medical sciences library setting examines the correlation of usage of a matched set of print and online titles, the validity of e-journals usage statistics and the impact of online journals on print journal usage. The print and online usage was determined for 270 journals, both versions of which were available. Print usage was determined annually since 1997 using the reshelving and the error-copies method. Online usage statistics were delivered by five publishers and corrected for redundant multiple accesses. Print journal usage decreased by 22.3 and 30.2% respectively over each of 2 years after the introduction of online journals. Journals published both in print and online lost 30.4% of their print usage within 2 years. The total loss of usage of print-only titles in the same period was somewhat higher, at 45.8%. The average correlation between online and print usage is 0.60 and 0.67 respectively. For the examined titles, users accessed the online versions ten times as often as the print version. Two clearly distinguishable groupings emerged: while with Academic Press and Elsevier, e-journal usage exceeded print usage by a factor of 3 or 4, the e journals of Blackwell, HighWire and Springer were used on average 14.6 times as frequently as the corresponding print journals. Each usage of a print article cost 2.79-50.82 Euro, each usage of an online article 0.31-15.10 Euro, depending on the publisher. On average, the usage of an online article was 5.4 times cheaper. Within 2-3 years the usage of online journals has outstripped that of print titles by a factor of ten, but the specific spectrum of usage remains much the same as when only print journals alone existed. Print titles not available online suffer a greater decline in usage compared with print/online journals. This confirms that what is read or purchased is determined primarily by ease of access and that there is a steady tendency to reduce the multiplicity of access modes to a manageable few. The availability of journals online seems to have created a new clientele, at least in the case of the German-language Springer journals. The connection between supplier and supply is much less clear with e journals than it is with print titles. Therefore it is very important to stress and encourage the role of the library as the supplier of this sort of information in the university environment. Collection building issues are discussed in the light of the results. PMID- 12641528 TI - Effectiveness of training health professionals in literature search skills using electronic health databases--a critical appraisal. AB - The objective was to assess the effect on health professionals' skills of one to eight hours literature search and retrieval training from electronic health databases. We searched: Cochrane library (2002; Issue 3), MEDLINE (1977-2002/5), EMBASE (1980-2002/7); CINAHL (1982-2002/5); ASSIA (1982-2002/7), BNI (1994 2002/5), ERIC (1985-2002/6); LISA (1969--current), NRR (2002, Issue 2), the world wide-web and references. The selection criteria consisted of randomised controlled trials, controlled before and after, and controlled cohort studies in comparison with no training. The intervention had to be one to eight hours training in literature search and retrieval skills for health professionals. The outcome was the effect on health professionals' literature search and retrieval skill levels measured through reliable instruments. For data collection and ana lysis, one reviewer extracted data and assessed the quality of the studies and the second reviewer checked it. The results indicate that there is some evidence of positive impact on health professionals' skill levels in literature searching and they find the training useful. In conclusion, the size of the positive effect is debatable as only three small and methodologically weak studies met the inclusion criteria and out of those only two showed the positive effect. PMID- 12641529 TI - Impact of a Graduate Entry Programme on a medical school library service. AB - The aim of this study was to compare the use of library facilities by first year undergraduate medical students and Graduate Entry Programme students (GEP). More specifically it tried to determine which library services (if any) were more frequently used by GEP so that this could be taken into account in future Information Services planning. A questionnaire on the use of Library and Information Services was posted to all first year GEP students and undergraduates on the 5-year course. In addition, user statistics of library entry and borrowing were collated from gate readings and the library Unicorn management system. Overall, GEP students were found to make a greater daily/weekly use of library facilities than undergraduates on the 5-year course. The facilities most used by both sets of students were essential texts, e-mail, PCs and study facilities. Computer Aided Learning packages, journals and video facilities were least used. However, on a daily/weekly basis GEP students made 74% more use of journals (P < 0.01), 59% more use of e-journals (P < 0.05), 36% more use photocopiers (P < 0.05), 42% more use of printers (P < 0.05), 56% more use of the library catalogue (P < 0.05) and 50% more use of databases (P < 0.05). This difference in use should be taken into account by LIS providers as there is expected to be an increase in fast-track graduate courses offered by medical schools throughout the UK. PMID- 12641530 TI - The Practice Nurse Project: a partnership approach to information provision in Leeds. PMID- 12641531 TI - A quest for questionnaires. PMID- 12641532 TI - Learning and teaching. PMID- 12641533 TI - Ghrelin and insulin metabolism. PMID- 12641534 TI - Complex disease genetics enters the high street. PMID- 12641535 TI - How to make clinical use of bacterial evasion strategies. PMID- 12641536 TI - Lack of association of the Glu298Asp polymorphism of endothelial nitric oxide synthase with manifest coronary artery disease, carotid atherosclerosis and forearm vascular reactivity in two Austrian populations. AB - OBJECTIVE: Conflicting data exists about the possible contribution of the homozygous Asp/Asp genotype of the Glu298Asp polymorphism of endothelial nitric oxide synthase to human atherosclerotic vascular disease. We investigated the polymorphism in two independent study populations: a case-control study including patients with angiographically verified coronary artery disease (CAD) on the one hand and a cross-sectional epidemiological study on the other hand. METHODS: The Glu298Asp polymorphism was determined by PCR-RFLP as established. In the case control study (240 patients and 248 controls) a possible association between the polymorphism and CAD, and age of onset of CAD and myocardial infarction was investigated. In the cross-sectional epidemiological study (932 subjects) intima media thickness (IMT) of the carotid artery as well as morphological plaque burden and forearm vascular reactivity (peak postischemic reactive hyperaemia, determined by venous occlusion plethysmography) were measured. RESULTS: In the case-control study genotype distribution (Glu/Glu; Glu/Asp; Asp/Asp) was not different between the CAD patients (43/46/11%) and the controls (49/41/10%, P = NS). No association of the polymorphism with age of onset of CAD or myocardial infarction was found. In the epidemiological study no influence of the genetic variant on IMT was observed after correction for classical determinants of IMT (average IMT: Asp/ Asp: 0.077 +/- 0.011 mm; Glu/Glu and Glu/Asp: 0.080 +/- 0.012 mm, P = NS). Forearm vascular reactivity was also not different between homozygous Asp/Asp subjects and Glu/Glu and Glu/Asp subjects (peak-reactive hyperaemia 20.1 +/- 7.3 mL min-1 100 mL-1 vs. 20.0 +/- 6.5 mL min-1 100 mL-1, P = NS). CONCLUSIONS: Our results suggest that there is no association of the Glu298Asp polymorphism with coronary or carotid atherosclerosis or forearm vascular reactivity in these populations recruited in a country with a rather high risk for atherosclerosis. We suggest additional investigations to be performed in populations at different risk for coronary events to further elucidate the possible contribution of this polymorphism to vascular disease. PMID- 12641537 TI - Cardiovascular responses to endurance training in children: effect of gender. AB - BACKGROUND: The aim of the present study was to determine in healthy children the effect of a well-controlled endurance training programme on cardiac function at maximal exercise and to define whether gender affects the training-induced cardiovascular response. The contribution of factors potentially involved in those adaptations such as cardiac dimensions and diastolic and systolic function was also investigated. METHODS: Thirty-five l0-11-year-old children took part in this study: 19 children (10 girls and nine boys) were assigned to participate in a 13-week endurance training programme (3 x 1 h week-1, intensity: > 80% HR max), and 16 (seven girls and nine boys) served as a control group. A resting echocardiographic evaluation and a maximal upright cycle test, including measurement of stroke volume (SV), cardiac output (Q) and blood pressure, were performed in all children before and after the study period. RESULTS: The training programme led to a rise in maximal O2 uptake (VO2max), brought about however, only by an increase in SVmax in both genders. Moreover, the boys increased their VO2max to a greater extent than the girls (boys: +15%; girls: +8%) only because of a higher SVmax improvement (boys: +15%; girls: +11%). No alterations were noticed in the SV pattern from rest to maximal exercise, indicating that the increase in SVrest was a key factor in the improvement of SVmax and thus VO2max. Regarding resting echocardiographic data, an increase in the left ventricular end-diastolic diameter, concomitant with an improvement in diastolic function, was observed after training and constituted an essential element in the rise in VO2max after training in these children. Moreover, during maximal exercise, a decrease in systemic vascular resistances, probably indicating peripheral cardiovascular adaptive changes, might also play an important role in the increase in VO2max. CONCLUSION: Whatever gender, aerobic training increases VO2max in children, mediated by an improvement in SVmax only. Similar mechanisms, including loading conditions and cardiac morphology, seem to be involved in both genders in order to explain such an improvement. PMID- 12641538 TI - Homocysteine-lowering treatment with folic acid plus vitamin B6 lowers urinary albumin excretion but not plasma markers of endothelial function or C-reactive protein: further analysis of secondary end-points of a randomized clinical trial. AB - BACKGROUND: Hyperhomocysteinaemia is an independent risk factor for atherosclerosis and is thought to induce its effects through causing endothelial dysfunction. We studied the effect of homocysteine-lowering treatment with folic acid plus vitamin B6 on urinary and plasma markers of endothelial function, and on plasma C-reactive protein, a marker of chronic inflammation. DESIGN: We performed a placebo-controlled 2-year trial among 158 healthy siblings of patients with premature atherosclerotic disease to determine the effect of daily folic acid (5 mg) plus vitamin B6 (250 mg) treatment as compared with placebo medication (n = 80) on markers of endothelial function (urinary albumin-to creatinine ratio and plasma concentrations of soluble E-selectin, soluble vascular cell adhesion molecule-1, von Willebrand factor, tissue-type plasminogen activator and plasminogen activator inhibitor-1) and inflammation (C-reactive protein). Outcome variables were assessed at baseline and after 1 and 2 years of treatment. RESULTS: Fasting homocysteine concentrations ( micromol L-1) at baseline and after treatment were 14.7 +/- 8.2 and 7.4 +/- 1.9 in the vitamin and 14.7 +/- 8.8 and 12.0 +/- 5.4 for the placebo group, respectively. Vitamin treatment was associated with a decreased urinary albumin-to-creatinine ratio at follow up [regression coefficient (beta) -0.20 mg mmol-1 (CI: -0.43-0.03); P = 0.09]. After adjustment for age, sex, baseline concentrations of postmethionine total homocysteine plus the baseline albumin-to-creatinine ratio, the beta was 0.23 mg mmol-1 (CI: -0.43 to -0.02; P = 0.03), which amounts to a decrease of approximately 20%. There was no apparent effect of vitamin treatment on the other markers. CONCLUSIONS: Homocysteine-lowering vitamin treatment in healthy siblings of patients with premature atherosclerotic disease is associated with a decreased urinary albumin-to-creatinine ratio, but not with other markers of endothelial dysfunction, or in plasma C-reactive protein. The clinical significance of these findings remains to be determined. PMID- 12641540 TI - Body mass index is the main risk factor for arterial hypertension in young subjects without major comorbidity. AB - BACKGROUND: Analytical statistics revealed a variety of risk factors for hypertension, but the complex interplay between different factors remains to be determined by more powerful statistical techniques. METHODS: Analytical as well as new, explorative statistical methods such as natural segmentation (k-means) and predictive modelling algorithms (C4.5) were used to classify the interactions of the individual risk factors for arterial hypertension in a large cohort of subjects. Fifty-five attributes (subject base, sociodemographic, medical history, laboratory data) were obtained from each of the 3547 participants of a community based health survey. The study subjects, mean age of 41 years, were free of major comorbidity. RESULTS: Twenty-five percent of the subjects had at least stage 1 hypertension. No clear linear dependency of risk factors with the diagnosis hypertension could be derived by the analytical statistics. In particular, the mutual amplification of different risk factors towards hypertension could not be revealed by these techniques. Explorative analytics however, uncovered body mass index (BMI) as the main single risk factor associated with hypertension. High predictive accuracy was achieved when combinations of certain risk factors including male gender and age were used. CONCLUSIONS: In summary, the survey of risk factors for hypertension using explorative analytics yielded high increases for the correct prediction of arterial hypertension. In this cohort, BMI was the single strongest parameter associated with arterial hypertension. PMID- 12641539 TI - Novel LPL mutation (L303F) found in a patient associated with coronary artery disease and severe systemic atherosclerosis. AB - BACKGROUND: Patients with lipoprotein lipase (LPL) deficiency had been generally thought to be spared accelerated atherosclerosis in spite of a marked elevation of plasma triglyceride levels. However, it has been recently reported that some heterozygous and homozygous LPL-deficient patients are associated with premature atherosclerosis. In this paper, we report a 55-year-old type I hyperlipidaemic patient with a novel missense mutation in the LPL gene. PATIENT AND RESULTS: The patient had suffered from coronary artery disease, abdominal aortic aneurysm, and stenoses of the bilateral renal arteries and superficial femoral arteries. Sequencing of the genomic DNA revealed that the patient was a homozygote for the mutation, a G to C transition at nucleotide position 1069 in the exon 6, resulting in an amino acid substitution of Phe for Leu303 (L303F). Approximately 6% and approximately 40% of normal LPL activity and LPL mass, respectively, were detected in the patient's postheparin plasma. An in vitro expression study demonstrated that COS7 cells transfected with L303F mutant cDNA produced a 40% amount of LPL protein in cell lysates compared with normal cDNA, but no protein was detected in the media. Lipoprotein lipase activity was completely absent in both lysates and media of the cells transfected with the mutant cDNA, suggesting that this mutation in the LPL gene results in the production of a functionally inactive protein. CONCLUSION: This case suggests that the LPL missense mutation (L303F), which impairs lipolysis but preserves the LPL mass, is proatherogenic. PMID- 12641541 TI - Effects of insulin lispro and chronic vitamin C therapy on postprandial lipaemia, oxidative stress and endothelial function in patients with type 2 diabetes mellitus. AB - BACKGROUND: Insulin therapy may influence cardiovascular disease (CVD) and lipid metabolism in type 2 diabetes (T2D). Exaggerated postprandial lipaemia (PPL) is a feature of diabetic dyslipidaemia affecting CVD via enhanced oxidative stress (OS) and endothelial dysfunction. We assessed endothelial function and OS during PPL following insulin and vitamin C. Twenty (17 M) T2D patients were studied (mean Hba1c 8.4%) at baseline, following 6 weeks of insulin lispro (0.2 Iu kg-1) and vitamin C 1-g daily. Eight-h lipid and glucose profiles were measured following a fatty meal. Endothelial function (flow-mediated vasodilatation: FMD) and OS were measured at fasting, 4 h and 8 h. MATERIALS AND METHODS: Glucose, body mass index, and total and LDL cholesterol remained unchanged. FMD improved. Placebo group: fasting, 1.1 +/- 1.2 to 4.2 +/- 1.1% (P < 0.001); 4-h, 0.3 +/- 1.2 to 3.1 +/- 0.9% (P < 0.01); 8-h, 0.7 +/- 1.1 to 3.76 +/- 1.1% (P < 0.001). Vitamin C group: fasting, 0.9 +/- 1.1 to 6.1 +/- 1.3% (P < 0.001); 4-h, 0.7 +/- 1.5 to 4.9 +/- 2.1% (P < 0.001); 8-h, 0.8 +/- 0.9 to 5.8 +/- 0.6% (P < 0.01). Post-prandial lipaemia was attenuated: TG area-under-curve (mmol L-1 8 h-1), 52.6 +/- 11 to 39.1 +/- 12.5 (placebo group), P < 0.02; and 56.9 +/- 8 to 40.1 +/- 10.3 (vitamin C group), P < 0.02. Oxidative stress was reduced, with greater changes in the vitamin C group. CONCLUSION: Insulin may thus exert vascular benefits in T2D, by modifying fasting and postprandial lipid metabolism resulting in reduced OS and improved EF. Vitamin C therapy may augment the vascular benefits of insulin in T2D through additional effects on OS and EF. PMID- 12641542 TI - Exocrine pancreatic dysfunction in sepsis. AB - BACKGROUND: Sepsis in critical illness is associated with the progressive failure of multiple organs. This study aims to establish a correlation between the severity of sepsis and exocrine pancreatic dysfunction. MATERIALS AND METHODS: In a prospective cohort study pancreatic exocrine function was tested by means of a secretin-cholecystokinin test in 21 critically ill, mechanically ventilated patients with sepsis according to criteria of the American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference Committee (ACCP/SCCM): 11 patients with shock and 10 patients without shock. Data were compared with seven healthy controls. RESULTS: The volume of duodenal fluid was not statistically different in the three groups. Sepsis patients without shock had significantly reduced content of amylase and chymotrypsin in duodenal juice compared with healthy controls (P < 0.01). Secretion of amylase, chymotrypsin, trypsin (P < 0.01 each) and bicarbonate in duodenal fluid (P < 0.05) was impaired in the septic shock patients when compared with the healthy controls. The content of trypsin was different between sepsis patients and septic shock patients (P < 0.05). Spearman correlation analysis was significant between the amylase secretion and the APACHE III and SOFA scores (P < 0.01). The SOFA score was also related to secretion of trypsin (P < 0.05). In patients on pressor therapy, use of norepinephrine was associated with a significant decrease in bicarbonate secretion (P < 0.05). CONCLUSIONS: Sepsis is associated with secretory pancreatic dysfunction that is worse in septic shock than in sepsis without shock. Impaired exocrine function was significantly correlated to the APACHE III and SOFA scores. PMID- 12641543 TI - Topical butyrate improves efficacy of 5-ASA in refractory distal ulcerative colitis: results of a multicentre trial. AB - BACKGROUND: The treatment of distal ulcerative colitis, refractory to conventional 5-ASA/steroid treatment, is still a matter of debate. The present study aimed at confirming, with adequate statistical power, previous data indicating the usefulness of topical butyrate and 5-ASA in the treatment of this condition. DESIGN: Double-blind, placebo-controlled, multicentre study. A total of 51 patients with distal (< 65 cm) ulcerative colitis, refractory to topical 5 ASA/cortisone, were randomly allocated to receive topical 5-ASA 2 g and 80 mM L-1 sodium-butyrate bid (Group A; 24 patients) or 5-ASA 2 g and 80 mL saline bid (Group B; 27 patients) for 6 weeks. Sigmoidoscopy with biopsies, as well as clinical and laboratory evaluations, were carried out at enrollment and at the end of the trial. Primary endpoints: remission or marked improvement in endoscopic, histologic and clinical findings. RESULTS: Most parameters showed a significant improvement vs. baseline in both groups. Remission in six patients and improvement in 12 patients in Group A vs. one remission and 13 with improvement in Group B (P < 0.05). A significant difference in favour of Group A was recorded regarding the number of bowel movements (P < 0.01), urgency (P < 0.05) and the patients' self evaluation (P < 0.01). DISCUSSION: The combined treatment with topical butyrate and 5-ASA is significantly more effective than 5 ASA alone in the management of refractory distal colitis. Further improvements in the treatment of refractory distal ulcerative colitis may be feasible based on the identification of patient subgroups and the association of two or more active drugs. Butyrate may well be one of them. PMID- 12641544 TI - Portal hypertension is associated with increased mRNA levels of vasopressor G protein-coupled receptors in human hepatic arteries. AB - BACKGROUND: The contractile response of human splanchnic vessels to different vasoconstrictors is attenuated in cirrhosis. Functional studies indicate a cellular signalling defect upstream of the G-protein level. The aim of the present study was to analyze expression and mRNA levels of the following most relevant vasopressor receptors in the smooth musculature of human hepatic arteries: alpha1 adrenoceptor (AR) subtypes a, b and d, angiotensin II type 1 receptor (AT1), arginine vasopressin receptor type 1a (V1a), endothelin receptor type A (ETA) and B (ETB). MATERIALS AND METHODS: Hepatic arteries were collected from 10 donors (noncirrhotic) and 14 recipients (cirrhotic) at liver transplantations. Real-time-PCR was performed to quantify steady-state levels of receptor mRNAs. RESULTS: alpha 1aAR mRNA levels showed no significant difference between the cirrhotic arteries and the controls while the mRNA levels of the other vasoactive receptors were significantly higher in the cirrhotic hepatic arteries (alpha 1bAR: 4-fold, P = 0.013; AT1: 16-fold, P = 0.024; V1a: 23-fold, P = 0.001; ETA: 4-fold, P = 0.02; ETB: 8-fold, P = 0.008). No mRNA for the alpha 1dAR was detected either in the donor or recipient hepatic arteries. CONCLUSION: We conclude that vascular hyporeactivity to the most relevant endogenous vasoconstrictors of cirrhotic hepatic arteries is not caused by a receptor down regulation at mRNA levels. In contrast they were up-regulated. PMID- 12641545 TI - Time to progression is dependent on the expression of the tumour suppressor PTEN in ovarian cancer patients. AB - BACKGROUND: Quantitative analyses of PTEN expression of ovarian cancer tissues were performed in this study. PTEN expression was investigated in terms of each patient's progression-free interval to indicate the role of PTEN in the generation of platinum refractory tumours. MATERIALS AND METHODS: The study group comprised 20 ovarian cancer patients from whom fresh frozen tissues of both the primary tumour and specimens of progressive disease were available. The PTEN protein and phosphorylation of the downstream effector protein kinase B (PKB) were quantified by Western blot analyses and subsequent densitometry. Data were analyzed for individual PTEN variation with respect to the clinical course as defined by the progression-free interval. RESULTS: Applying the usual clinical criteria for platinum-sensitivity after progression, seven patients were considered platinum-sensitive whereas 13 patients had suffered a progression within 12 months after the chemotherapy. In 5/7 (71%) cases with prolonged time to progression, an increase in PTEN was observed. Decline of PTEN expression occurred in 9/13 (69%) patients with poor outcome. PTEN expression corresponds inversely to PKB phosphorylation in 14/20 (70%) tissues investigated. CONCLUSIONS: The data suggest that decreased PTEN expression accompanies the progression of ovarian cancer. Declining PTEN expression results in a shortened relapse-free interval, whereas an increase of PTEN prolongs the time to progression. However, as far as recurrence occurs, PTEN is not the only mechanism to suppress tumour progression in ovarian cancer. PMID- 12641546 TI - Quantitative determination of MDR1 mRNA expression in peripheral blood lymphocytes: a possible role of genetic polymorphisms in the MDR1 gene. AB - BACKGROUND: P-glycoprotein is a transmembrane efflux pump that extrudes a wide variety of drugs, thereby reducing their intracellular access. In humans, P glycoprotein is encoded by the MDR1 gene. Recently, several single nucleotide polymorphisms in the MDR1 gene were identified. Moreover, it was postulated that, in addition to the full-length P-glycoprotein, a 'mini' P-glycoprotein was also present in lymphocytes. MATERIALS AND METHODS: We investigated the effect of the genetic polymorphisms G2677T and C3435T in the MDR1 gene on MDR1 mRNA expression in FACS-sorted peripheral blood CD4+, CD8+, CD19+, and CD56+ cells. MDR1 mRNA expression was determined in 45 healthy individuals using a real-time quantitative RT-PCR. RESULTS: We detected the highest expression of MDR1 mRNA in CD56+ cells, followed by CD8+ > CD4+ > CD19+ cells. However, genetic polymorphisms of the MDR1 gene failed to affect (P > 0.05) MDR1 mRNA levels in the peripheral blood lymphocytes. Furthermore, the transcript levels for the MDR1 N-terminal half were almost two-fold lower than that of the MDR1 C-terminal half in all cell populations investigated (P < 0.0001). CONCLUSIONS: An almost two fold difference in MDR1 C- and N-terminal half expressions supports the presence of mini-P-glycoprotein, an alternatively spliced form of the full-length molecule, in peripheral blood lymphocytes. PMID- 12641547 TI - Homocysteine during the menstrual cycle in depressive women. AB - BACKGROUND: Two possible factors that may have a causal relation with both depressive disorder and cardiovascular disease are elevated homocysteine and steroid hormones. Our previous study found significant changes in the plasma homocysteine concentration during the menstrual cycle in healthy women. The purpose of this study therefore was to test homocysteine in depressive women treated with fluoxetine during the menstrual cycle. MATERIALS AND METHODS: Thirteen premenopausal women suffering from mixed anxiety-depressive disorder and a control group of 15 healthy women were enrolled in this study. The homocysteine concentration was determined by high-performance liquid chromatography with fluorescence detection, and estradiol, progesterone and cortisol by RIA methods. RESULTS: We found significantly higher plasma homocysteine concentrations in the follicular phase than in the luteal phase of the menstrual cycle in both the depressive group (P < 0.003) and the controls (P < 0.0009). Moreover, the patient values of total homocysteine were significantly higher in the follicular phase (P < 0.03) and also in the luteal phase (P < 0.007) than the values of the controls. Estradiol and cortisol were significantly higher in the follicular phase of the patients compared with the control group. CONCLUSION: According to our results, women suffering from mixed anxiety-depressive disorder have not only significantly different concentrations of homocysteine in the follicular and luteal phase of the menstrual cycle but also higher plasma homocysteine compared with healthy women. More elevated homocysteine in the depressive than in the healthy premenopausal women points to the notion that psychological factors might be important when considering the homocysteine concentration. PMID- 12641551 TI - The development of cancer services: reflections on a learning journey. PMID- 12641548 TI - Elevated exhalation of hydrogen peroxide in patients with systemic sclerosis. AB - BACKGROUND: Systemic sclerosis is accompanied by an influx of activated phagocytes into distal airways. These cells release H2O2, which may evaporate from the airways surface and be detected in expired breath condensate. We tested whether patients with systemic sclerosis exhale more H2O2 than healthy subjects and whether breath condensate H2O2 levels correlate with some clinical parameters. MATERIAL AND METHODS: H2O2 was measured fluorimetrically in the expired breath condensate of 27 patients (22 women, five men, mean age 49 +/- 13.1 years) with systemic sclerosis and 27 age- and sex- matched healthy controls. RESULTS: Exhaled H2O2 levels were 3.5-fold higher (0.88 +/- 0.62 microM vs. 0.25 +/- 0.17 microM, P < 0.001) in the patients with systemic sclerosis than in the controls. Treatment with cyclophosphamide and/or prednisone (29 +/- 50 months, range 3-168 months) did not significantly decrease H2O2 exhalation (0.78 +/- 0.50 microM, n= 10 vs. 0.94 +/- 0.67 microM, n= 17, P > 0.05). No significant difference was found between patients with limited and diffuse scleroderma (1.03 +/- 0.69 microM, n= 17 vs. 0.63 +/- 0.41 microM, n= 10, P > 0.05). H2O2 levels correlated with disease duration (r = 0.38, P < 0.05) and time from the first Raynaud's episode (r = 0.44, P < 0.05). CONCLUSIONS: Patients with systemic sclerosis exhale more H2O2 than healthy controls, suggesting involvement of reactive oxygen species in disease processes. Lack of significant intergroups differences in H2O2 levels may have resulted from the small number of patients analyzed. PMID- 12641552 TI - A survey exploring the educational needs of care practitioners in learning disability (LD) settings in relation to death, dying and people with learning disabilities. AB - This paper presents the findings of a small scale pilot study which explored the educational base and needs of qualified care practitioners in Learning Disability (LD) settings in relation to death, dying and people with learning disabilities. Eighty questionnaires were sent to two NHS Trusts in the South of England. The response rate for the qualified care practitioners from Cherry Blossom (CB) was 100%, whereas for Greengages (GG), the response rate was only 25%. The response from the unqualified care practitioners was disappointingly low, hence we declared them null and void. The analysis of data highlighted major concerns: namely, a lack of consistent policy in the recording of death in residential homes for dying persons with LD; a lack of knowledge, particularly in psychosocial aspects and skills in care of dying persons. The majority of the qualified care practitioners surveyed highlighted the importance of communication with clients and their families. We recommend that communication and interpersonal skills in the care and management of the terminally ill persons with LD be the core component in the nursing curriculum which at present only indicates a trace of it. It is not made explicit that it is essential. This study supports the notion that issues of LD override and obscure physical illness. Our study also highlights ambiguity in the use of concepts and terminology, and demonstrates some limitations in our methodology. We propose that further research, using different methodological approaches, such as Ethnography, Ethnomethodology, or a combination of these, would be appropriate. PMID- 12641553 TI - Cancer-related fatigue--a difference of opinion? Results of a multicentre survey of healthcare professionals, patients and caregivers. AB - The objective of this study was to investigate the perceptions of patients with cancer, their caregivers and healthcare professionals (HCPs) about fatigue and its impact on quality of life. It was a cross-sectional survey, the respondents were patients with cancer attending three UK regional cancer centres (n = 1,370), their informal caregivers (n = 1,370) and a random selection of HCPs (oncologists/nurses/radiographers/haematologists; n = 1,098). The response rates for patients, caregivers and HCPs were 42%, 33% and 34% respectively. Fatigue was reported to affect 56% of patients and to have a considerable impact on quality of life. Caregivers also recognized that fatigue was a common problem, with significant effects on patients' quality of life and impact on themselves. Healthcare professionals recognized that fatigue was a common problem for their patients but overestimated its impact on some aspects of patients' daily lives. Although most HCPs reported that they prescribed/recommended treatment for over half of their patients, only 14% of patients reported receiving any such treatment. The most common advice was to take more rest and relaxation. CONCLUSIONS: patients with cancer report that fatigue is a common and distressing symptom and the importance of this symptom is generally recognized by both HCPs and lay-carers. Healthcare professionals need more information about the effectiveness of existing interventions for cancer-related fatigue and further research is required to improve the current management of this debilitating symptom. PMID- 12641554 TI - Italy reforms national policy for cancer pain relief and opioids. AB - Treatment of pain caused by cancer in Italy has been reported to be inadequate for more than a decade. The problem has been documented in the literature by International Narcotics Control Board (INCB) reports that show Italy's low consumption of morphine, and by INCB statements reflecting concern that pain medications are not adequately available to suffering cancer patients. The reasons for undertreatment include lack of physician education, low public awareness about pain management and overly restrictive regulations that inhibit the prescribing of opioid analgesics. In 1999, a workgroup appointed by the Ministry of Health began to address inadequate cancer pain management. In 2000, the workgroup issued a proposal to reform national policy, including simplification of opioid prescribing requirements. The proposal was adopted by Parliament in January 2001 and was effective from March 6th. Distribution of a new opioids prescription form to selected regions in Italy began in December 2001. The change, when fully implemented, will be an important step toward relieving cancer pain. PMID- 12641555 TI - Do newly diagnosed lung cancer patients feel their concerns are being met? AB - Lung cancer is the leading cause of cancer death worldwide. It has a poor prognosis and the majority of those affected are elderly. Evidence suggests that providing clear, relevant information and addressing patients' concerns can make a worthwhile difference to patients. This study aimed to: explore the concerns of lung cancer patients shortly after diagnosis; and enquire whether these concerns had been discussed by their care teams. Eighty patients with a new diagnosis of primary lung cancer were interviewed 14-28 days after the date on which they were told the diagnosis. Interviews were conducted either in the hospital ward, outpatient clinic or at home. Participants were asked to rate 17 specific items of concern from 1 = 'not a worry' to 5 = 'extremely worried', plus one non specific item. Patients rated at least two items as worrying them to some degree with a median of nine concerns being reported. Major concerns for patients were the illness itself; the future relating to the illness and concerns about the family. Overall, patients in the study felt that less than half of their concerns (43%) had been discussed by the care team. Although levels of concern about physical symptoms were relatively low, these had been more frequently addressed than the psychosocial issues, which were rated higher by patients. There were some differences in the number of concerns reported between males/females and younger/older age groups, but the pattern of concerns was similar. There were no differences in the level of concerns between treatment groups; the location of the interview nor in the interval between diagnosis and delivery of the checklist. This study supports previous findings that there is a need for health professionals to provide emotional support and respond to the psychosocial needs of patients by eliciting their concerns and attempting to address them in the early stages of the disease process. PMID- 12641556 TI - Distress, quality of life and strategies to 'keep a good mood' in patients with carcinoid tumours: patient and staff perceptions. AB - Patients with carcinoid tumours have reported a relatively good health-related quality of life (HRQoL) (European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-C30, EORTC QLQ-C30), and low levels of anxiety and depression (Hospital Anxiety and Depression Scale, HADS). The aim was to test the validity of these results. Data were gathered through interviews with 19 patients and 19 staff. Participants were asked about disease and treatment related distress, important aspects of quality of life and strategies to 'keep a good mood'. Patients were interviewed about themselves and staff were interviewed about a certain patient. Data were analysed by content analysis. Identified aspects of distress and quality of life were referred to an emotional, a physical and a social dimension. Most aspects of distress were of a physical character whereas most aspects of quality of life were of a social character. Several aspects of emotional distress not included in the EORTC QLQ-C30 and/or the HADS were identified. PMID- 12641557 TI - Validation of the French 'multidimensional fatigue inventory' (MFI 20). AB - Fatigue has become an important symptom in research and also for clinical diagnosis and follow up. Many physical illnesses, in particular chronic ones such as cancer, are highly associated with fatigue. Various questionnaires for measuring fatigue have been developed, but currently no validated questionnaire exists in French language. We selected the 'Multidimensional Fatigue Inventory' (MFI) which has been validated in its English version and then translated into French by the designers. This study describes the validation step of the French version of MFI and presents the psychometric properties of this instrument. A sample of 225 patients was divided into three groups 'Tired' (82 subjects), 'Moderately tired' (36) and 'Not tired' (107). The analysis of the structure validity found four dimensions: 'General Fatigue', 'Mental Fatigue', 'Reduced Activities' and 'Motivation'. The convergent validity showed highly significant correlation (P < 0.001) with a visual analogue scale. The French MFI has been able to distinguish patients with different fatigue levels. Cronbach's alpha measurement of the scale and the subscales are good, cronbach alpha > 0.70. The reproducibility and sensitivity to change in patients who were followed up one month later show satisfactory results. This validation study of the French version of MFI shows that this instrument is valid for clinical application and allows different dimensions of fatigue to be assessed which is of particular interest for physicians, especially for cancer carers, where fatigue assessment is an important aspect of the follow up. PMID- 12641558 TI - Young people with cancer--how should their care be organized? AB - The objective of this study was to assess the attitudes and views of young people receiving treatment for cancer, along with the views of their parents, their healthy peers and those involved in treating them without a cancer centre. The study analysed qualitative data collected using focus groups and patient interviews. Fourteen patients aged between 15 and 20 years were interviewed at a major cancer centre in Yorkshire. A focus group of healthy adolescents was held at a location away from the hospital to gain the perspectives of healthy adolescents. No disadvantages of a more centralized model of care for adolescents were discovered. The particular feature felt to be important was the expertise of the staff rather than the physical environment. Some suboptimal care was identified in referring units, though this appeared to apply to that associated with the treatment of solid tumours rather than leukaemias. The case for a more centralized approach to the management of young people with cancer is confirmed, and supports the model already implemented in the care of children with cancer. PMID- 12641559 TI - Symptom distress, functional status and health-related quality of life before high-dose chemotherapy with stem-cell transplantation. AB - The aims of this study were to describe how a group of patients with different malignant diseases perceived symptom distress (SD), functional status (FS) and health-related quality of life (HRQOL) on admission to the hospital for stem-cell transplantation (SCT), to compare the obtained data regarding FS and HRQOL with similar data from two general-population groups, and to relate the results to disease- and treatment-specific data. Fifty-one patients participated in the study. Three instruments were used to collect data: SFID-SCT, SIP and SWED-QUAL. The majority of the patients (92%) reported ongoing symptoms even before the SCT with tiredness (67%) and anxiety (53%) as the two most commonly reported symptoms. Although tiredness and anxiety were reported to be the most frequently occurring symptoms, these symptoms were not considered to cause that much distress. Instead, vomiting, reduced mobility and fever, although less commonly occurring, were reported as highly distressing when present. Compared with the general-population groups, the patients reported significantly poorer FS and HRQOL but no statistically significant correlations were found between SD, FS or HRQOL and the time since the last chemotherapy cycle or cycles respectively. Patients with advanced disease and patients with multiple myeloma were found to report more SD and poorer FS and HRQOL. PMID- 12641560 TI - Reports of information and support needs of daughters and sisters of women with breast cancer. AB - The aim of this study was to describe the information and support needs of women who have primary relatives with breast cancer. The Information and Support Needs Questionnaire (ISNQ) was developed and revised from previous qualitative and pilot studies. The ISNQ addressed concepts of the importance of, and the degree to which, 29 information and support needs related to breast cancer had been met. The study sample consisted of 261 community-residing women who had mothers, sisters, or a mother and sister(s) with breast cancer. Data were collected using a mailed survey. In addition to the ISNQ, additional items addressed family and health history, breast self-care practices, perception of the impact of the relative's breast cancer and other variables. Also included were established and well-validated measures of anxiety and depression. The findings document women's priority information and support needs. The information need most frequently identified as very important was information about personal risk of breast cancer. Other highly rated needs addressed risk factors for breast cancer and early detection measures. Generally, the women perceived that their information and support needs were not well met. These findings illuminate needs of women for more information and support when they have close family relatives with breast cancer and opportunities for primary care providers to assist women in addressing their needs. PMID- 12641561 TI - An evidence-based specialist breast nurse role in practice: a multicentre implementation study. AB - The objective of this study was to examine the feasibility, implementation, acceptability and impact of an evidence-based specialist breast care nurse (SBN) model of care in Australia. Primary data were collected from four diverse Australian breast cancer treatment centres over a 12-month period. The design was a multicentre demonstration project. Information about the provision of care and patient needs was collected through prospective logs. Structured interviews were conducted with women who received the SBN intervention (N = 167) and with a control group of women treated prior to the intervention period (N = 133). Health professionals (N = 47) were interviewed about their experience of the SBN. Almost all women had contact with an SBN at five scheduled consultations and 67% of women in the intervention group requested at least one additional consultation with the SBN. Women in the intervention group were more likely to receive hospital fact sheets and to be told about and participate in clinical trials. Ninety-eight per cent of women reported that the availability of an SBN would affect their choice of hospital, with 48% indicating that they would recommend only a hospital with a SBN available. Health professionals reported that SBNs improved continuity of care, information and support for the women, and resulted in more appropriate referrals and use of the time of other members of the team. In conclusion, the SBN model is feasible and acceptable within diverse Australian treatment centres; there is evidence that some aspects of care were improved by the SBN. PMID- 12641564 TI - Epigenetic reprogramming in mammalian nuclear transfer. AB - With the exception of lymphocytes, the various cell types in a higher multicellular organism have basically an identical genotype but are functionally and morphologically different. This is due to tissue-specific, temporal, and spatial gene expression patterns which are controlled by genetic and epigenetic mechanisms. Successful cloning of mammals by transfer of nuclei from differentiated tissues into enucleated oocytes demonstrates that these genetic and epigenetic programs can be largely reversed and that cellular totipotency can be restored. Although these experiments indicate an enormous plasticity of nuclei from differentiated tissues, somatic cloning is a rather inefficient and unpredictable process, and a plethora of anomalies have been described in cloned embryos, fetuses, and offspring. Accumulating evidence indicates that incomplete or inappropriate epigenetic reprogramming of donor nuclei is likely to be the primary cause of failures in nuclear transfer. In this review, we discuss the roles of various epigenetic mechanisms, including DNA methylation, chromatin remodeling, imprinting, X chromosome inactivation, telomere maintenance, and epigenetic inheritance in normal embryonic development and in the observed abnormalities in clones from different species. Nuclear transfer represents an invaluable tool to experimentally address fundamental questions related to epigenetic reprogramming. Understanding the dynamics and mechanisms underlying epigenetic control will help us solve problems inherent in nuclear transfer technology and enable many applications, including the modulation of cellular plasticity for human cell therapies. PMID- 12641565 TI - Expression of matrix metalloproteinase 8 (MMP-8) and tyrosinase-related protein-1 (TYRP-1) correlates with the absence of metastasis in an isogenic human breast cancer model. AB - The multi-step nature of metastasis poses difficulties in both design and interpretation of experiments to unveil the mechanisms causing the process. In order to facilitate such studies, we have previously derived a pair of breast tumor cell lines that originate from the same breast tumor but which have diametrically opposite metastatic capabilities. In this system, the monoclonal cell line M-4A4 is metastatic to the lungs of athymic mice, whereas clone NM-2C5 is equally tumorigenic but non-metastatic. Here, we report that representational difference analysis (RDA) of cDNA obtained from the two clonal populations revealed an increased expression of tyrosinase-related protein-1 (TYRP-1) and the matrix metalloproteinase-8 (MMP-8) genes in the non-metastatic cell line. RNA and protein analyses in cultured cells and in primary xenograft tissues confirmed that the non-metastatic cell line expresses TYRP-1 and MMP-8 at levels that are at least 20-fold higher than the metastatic counterpart. Other members of the MMP family (MMP-9 and MMP-2) and the tissue inhibitor of metalloproteinase-2 (TIMP-2) were found to be expressed at similar levels in both populations. The effects of MMP-8 and TYRP-1 on in vitro invasion and migration were assessed in cells whose expression of these genes was altered by stable transduction with sense and antisense constructs. Specific down-regulation of MMP-8 in non-metastatic NM-2C5 cells resulted in a 2.5-fold increased capacity to invade through Matrigel. Unlike other members of the matrix metalloproteinase family, MMP-8 has not previously been implicated in the processes of tumorigenesis or metastasis. The successful identification of two proteins that are differentially expressed in these matched clonal cell lines and the tumors that they produce demonstrates the feasibility of using this approach to search for genes that are associated with aberrant differentiation toward metastatic behavior. PMID- 12641566 TI - Increased levels of the translation initiation factor eIF4E in differentiating epithelial lung tumor cell lines. AB - Rates of eukaryotic protein synthesis and proliferation are dependent upon the availability of eIF4F, the cap-binding translation initiation complex that guides the ribosome onto the mRNA. One possible rate-limiting factor in eIF4F complex formation is the availability of eIF4E, which interacts specifically with the mRNA cap structure. As such, it has a potential role in the selective translation of growth-related mRNAs, with overexpression of eIF4E resulting in aberrant cell growth and transformation. A number of studies suggest that eIF4E may play a role in cellular differentiation as well as proliferation. We have previously reported that post-transcriptional regulation is involved in the induction of keratins in epithelial lung tumor cell lines exposed to the differentiation-modulating agent, bromo-deoxyuridine (BrdU). Here, we demonstrate that these BrdU-treated lung cells express elevated levels of eIF4E protein and enhanced phosphorylation of eIF4E. Overexpression of eIF4E by cDNA transfection in the poorly differentiated, keratin-negative human lung cell line, DLKP, was found to promote a flattened, more epithelial appearance to these cells, coupled with the induction of simple keratins (keratins 8 and 18). In contrast, levels of eIF4E expression were found to decrease during BrdU-induced differentiation of the leukemic cell line, HL-60, suggesting that there are cell-type differences in the response to BrdU and in the requirement for eIF4E during differentiation. PMID- 12641567 TI - Developmental fate of embryonic germ cells (EGCs), in vivo and in vitro. AB - Embryonic germ cells (EGCs) derived from mouse primordial germ cells (PGCs) are known both to colonize all cell lineages of the fetus and to make tumors in vivo. When aggregated with eight-cell embryos, EGCs from a new EGC line expressing green fluorescent protein (GFP) were found to contribute preferentially to the epiblast but unexpectedly were also capable of colonizing primary endoderm. When injected under the kidney capsule, EGCs derived from 12.5 days post coitum (dpc) PGCs formed differentiated tumors. The ability of EGCs to differentiate in an organ culture system depends upon their partners in cell culture. When EGCs, marked with a LacZ transgene, were mixed with disaggregated and reaggregated mouse fetal lung in an organ culture system, they remained undifferentiated. In urogenital ridge reaggregates on the other hand, some EGCs were capable of differentiating to form small epithelial cysts. PMID- 12641568 TI - The Balbiani body and generation of early asymmetry in the oocyte of a tiger beetle. AB - The developmental changes within the Balbiani body in previtellogenic and early vitellogenic oocytes of a tiger beetle, Pseudoxycheila angustata, are described. Our study showed that the Balbiani body forms in a juxtanuclear position in previtellogenic oocytes. Subsequently, it disperses within the ooplasm while multivesicular bodies, a prominent component of the Balbiani body in this species, segregate out and are targeted to the posterior pole of the oocyte. We demonstrated that the Balbiani body is a temporary site of organelle accumulation and sorting and it is involved in the creation of an early polarity during oogenesis. Our data suggest that the multivesicular bodies, initially associated with the Balbiani body, may ultimately contribute to the formation of the pole plasm (oosome). Our study is the first description of the presence of the Balbiani body in oocytes of an insect with a meroistic ovary and only the second known example of the Balbiani body in insects in general. In addition, we showed, for the first time, that the components of Balbiani body participate in the formation of the pole plasm in insects. Interestingly, the oocytes of a European species of tiger beetles do not develop the Balbiani body. We discuss the developmental and evolutionary aspects of this finding. PMID- 12641569 TI - An expression pattern screen for genes involved in the induction of the posterior nervous system of zebrafish. AB - The posterior nervous system, including the hindbrain and the spinal cord, has been shown to be formed by the transformation of neural plate of anterior character by signals derived from non-axial mesoderm. Although secreted factors, such as fibroblast growth factors (FGFs), Wnts, retinoic acid (RA) and Nodal, have been proposed to be the posteriorizing factors, the mechanism how neural tissue of posterior character is induced and subsequently specified along the anteroposterior axis remains elusive. To identify intercellular signaling molecules responsible for posteriorization of the neural plate as well as to find molecules induced intracellularly by the posteriorizing signal in the caudal neural plate, we screened by in situ hybridization for genes specifically expressed in posterior tissues, including the posterior neural plate and non axial mesoderm when posteriorization of the neural plate takes place. From a subtracted library differentiating anterior versus posterior neural plate, 420 cDNA clones were tested, out of which 76 cDNA fragments showed expression restricted to the posterior tissue. These clones turned out to represent 32 different genes, including one novel secreted factor and one transmembrane protein. Seven genes were induced by non-axial mesodermal implants and bFGF beads, suggesting that these are among the early-response genes of the posteriorizing signal. Thus, our approach employing cDNA subtraction and subsequent expression pattern screening allows us to clone candidate genes involved in a novel signaling pathway contributing to the formation of the posterior nervous system. PMID- 12641570 TI - Endoderm- and mesenchyme-dependent commitment of the differentiated epithelial cell types in the developing intestine of rat. AB - During organogenesis, the intestinal tract progressively acquires a functional regionalization along the antero-posterior axis. Positional information needed for enterocytes has been studied, but the mechanisms that control Paneth and endocrine cell differentiation are poorly understood. We have used a model of endoderm/mesenchyme cross-associations to evaluate the respective roles of endoderm and mesenchyme in the cytodifferentiation of these epithelial cells. Heterotopic cross-associations comprising endoderm and mesenchyme from the presumptive proximal jejunum and colon were developed as xenografts in nude mice. Our results show that endoderm from the presumptive proximal jejunum when associated with colonic mesenchyme generate small intestinal enterocytes. Interestingly, no lysozyme-producing cells were generated. On the other hand, associations comprising colon endoderm and jejunal mesenchyme showed heterodifferentiation with typical small intestinal morphology with sucrase isomaltase expression and Paneth cell differentiation. Heterotopic associations developed enteroendocrine cell patterns according to the normal fate of the endodermal moiety. As enteroendocrine cell commitment seems to occur before the other intestinal cell types, we cannot exclude a role of instructive signals from the mesenchyme on endocrine cell differentiation earlier in the development. These results identified a complex pattern of cell commitment, dependent of the differentiation type of the epithelial cell, on the regional origin of the endoderm and the associated mesenchyme. PMID- 12641571 TI - Contact sensitization to N-(cyclohexylthio)phthalimide. AB - Previous reports revealed relative high sensitization rates to the rubber chemical N-(cyclohexylthio)phthalimide (CTP; CAS-No. 17796-82-6), but the relevance of positive reactions remained unknown. It was discussed whether the test concentration of 1% pet. needed to be changed. The German Contact Dermatitis Research Group (DKG) added CTP in 3 concentrations, i.e. 0.25% pet., 0.5% pet, 1% pet., to the rubber series. From June 1999 to December 2000, 1936 patients in 30 departments of dermatology were tested with CTP. Of the 56 patients with a positive test reaction (2.9%), 52 reacted to CTP 1% pet., 21 to CTP 0.5% pet., and 9 to CTP 0.25% pet. The reaction indices were about the same with all concentrations. 34 patients with a positive reaction to CTP 1% pet. did not react to the lower concentrations. The majority of these reactions are probably false positive. With CTP 0.25% pet., however, the majority of true allergic reactions to CTP were missed. Analysis of population characteristics and concomitant sensitizations to other rubber chemicals led to the conclusion that a positive reaction to CTP 0.5% pet. was a good indicator of contact allergy to CTP. Thus, the DKG decided to continue patch testing with CTP 0.5% pet. in the rubber series. Manufacturers' information about the use of CTP seem to partly contradict the patients' characteristics seen in this study. So the relevance of positive CTP patch test reactions, and the causative exposures in patients with CTP allergy, still remain to be clarified. PMID- 12641572 TI - Hand eczema in 45 bakers - a clinical study. AB - The skin of bakers is heavily exposed to dough, spices, water and detergents. This is follow-up of a previous Swedish questionnaire study showing bakers to have a 3-fold increased risk of hand eczema. The aims were to establish diagnoses and to study consequences of hand eczema. From a cohort study of 2226 bakers, a random sample among bakers reporting hand eczema was examined. Of 60 randomly selected bakers reporting a history of hand eczema, 52 attended an examination comprising a standardized interview, documentation of clinical skin signs, patch testing and prick testing with standard and bakery series, and serum analyses. In all, 45 bakers confirmed a history of hand eczema, for which 11 (24%) had been on sick leave, with a median duration of 14 weeks. 13 (29%) had changed their occupation due to skin disease, 19 had positive patch test reactions to standard contact allergens - and 5 to bakery contact allergens. 16 bakers had positive prick tests to standard allergens, 10 to bakery allergens, of whom 9 reacted to flours. Since considerable medical and social consequences of hand eczema are seen, thorough diagnosis of contact allergy and IgE-mediated allergy in bakers, as well as preventive measures, are essential. PMID- 12641573 TI - Primary oxidation products affect the quantification of formaldehyde in autoxidized fatty alcohol ethoxylates when using DNPH derivatization. AB - Non-ionic surfactants, e.g. fatty alcohol ethoxylates, are considered to cause less skin irritation than other types of surfactants. However, the autoxidation of alcohol ethoxylates generates products that are both skin irritating and sensitizing, such as formaldehyde. It has been suggested that formaldehyde be used as an indicator of the degree of autoxidation of fatty alcohol ethoxylates, and thus also as an indicator of the sensitizing capacity of the product. Attempts to analyse formaldehyde in autoxidized ethoxylates using 2,4 dinitrophenylhydrazine derivatization resulted in formation of the corresponding hydrazone not only from free formaldehyde but also from autoxidation products during the derivatization reaction. Consequently, this derivatizing agent is inappropriate for the determination of formaldehyde in autoxidized fatty alcohol ethoxylates. Focus on the content of primary oxidation products, i.e. hydroperoxides/peroxides, is suggested, particularly as they also are potent sensitizers. PMID- 12641574 TI - Erythema multiforme due to contact with weeds: a recurrence after patch testing. AB - Erythema multiforme (EM) as a complication of patch testing (PT) is rare. A 52 year-old woman with a 13-year history of episodes of EM, after contact with weeds during home gardening, had had no recent history of herpes simplex, other infection, drug ingestion or vaccination. On examination, EM lesions were distributed on the exposed skin. 5 weeks after complete resolution, PT and photopatch testing (PPT) were done with fresh plants she brought in. She was PT with a standard series and the Hermal-Trolab plants, woods, tars, balsams and flavors series. Intradermal testing, with a 3 + reaction to mixed weed pollens, was done 3 weeks later. Specific IgE to weed pollens class 1 (CAP-Pharmacia) was detected. Eczematous PT reactions were obtained with fresh leaves: common chickweed (Stellaria media Caryophyllaceae), dandelion (Taraxacum officinale Compositae), field-milk thistle (Sonchus arvensis Compositae) and white clover (Trifolium repens Leguminosae). Photoaggravation was seen to common chickweed and dandelion. Positive PT was also seen with alantolactone. By the 4-day reading, a typical EM had commenced, coming up to quite the same extent as seen on admission. There was no photosensitivity (UV skin tester, K. Waldmann). In the essential oil obtained from common chickweed, thin layer chromatography (TLC) revealed the well-known contact allergens borneol, menthol, linalool, 1,8 cineole, and other terpenes such as epoxy-dehydro-caryophyllene, monoterpene alcohol-ester and caryophyllene. Up to now, no data on essential oil in Stellaria media (common chickweed) have been reported. It can be concluded that EM developed due to contact with weeds, and recurred after patch testing. Neither blistering nor eczematous lesions have been seen on her skin, making this case very unusual. As far as the world literature is concerned, this is only the 4th report of EM developing in association with patch testing. PMID- 12641575 TI - Profile of irritant patch testing with detergents: sodium lauryl sulfate, sodium laureth sulfate and alkyl polyglucoside. AB - The cutaneous reaction to detergents follows distinct kinetic rules: the duration of application and the irritant concentration are of major importance. The aim of this study was to evaluate the differences in kinetics of skin reaction between the standard irritant sodium lauryl sulfate (SLS), and 2 modern detergents: sodium laureth sulfate (SLES) and alkyl polyglucoside (APG). We performed patch testing with SLS and SLES (or APG) at different concentrations (0.125, 0.25, 0.5, 1.0 and 2.0%) and with different exposure times (6, 12 and 24 h). Evaluation was conducted by measurement of transepidermal water loss (TEWL) and laser Doppler flowmetry (LD) 24 h, 7 and 10 days after patch removal. We found a pronounced reaction to SLS, and a far milder one to SLES. Even at the highest concentration the skin reaction to APG was hard to detect. During the regeneration period (day 3-10) SLS showed even at day 10 an increased TEWL at all concentrations tested. The irritation due to SLES was convincingly detectable only up to day 7, whereas the APG-tested skin areas showed no significant reaction even at day 3. These results demonstrate the improvement in reduction of skin irritation achieved by development of novel detergents. PMID- 12641576 TI - Risk factors for contact allergy to nickel - results of a multifactorial analysis. AB - To quantify the independent impact of potential risk factors for nickel contact allergy (NCA), a multifactorial Poisson regression analysis of standardized anamnestic and patch test data (with nickel sulfate, 5% in petrolatum) was performed, comprising 74 940 patients assessed in the 33 German and Austrian contact dermatitis units of the Information Network of Departments of Dermatology (IVDK) between 1992 and 2000. NCA was observed in 15.5% patients. Female sex was the strongest risk factor (prevalence ratio 3.74, 95% CI: 3.51-3.98). Risk increased monotonically and significantly with decreasing age. Atopic dermatitis was not a risk factor. The year of patch test had no influence on NCA risk. Significant variation of risk between occupations was observed. In conclusion, our multifactorial analysis was able to quantify the impact of established risk factors and additionally address other, e.g. occupational, factors yet unidentified. PMID- 12641577 TI - Occupational rubber glove allergy: results of the Information Network of Departments of Dermatology (IVDK), 1995-2001. AB - About 21% of the patients with occupational contact dermatitis registered in the Information Network of Departments of Dermatology (IVDK) in the years 1995-2001 were patch tested due to suspected rubber glove allergy. We analysed reaction frequencies to thiurams, dithiocarbamates, mercaptobenzothiazole (MBT) and its derivatives, thioureas, and 1,3-diphenylguanidine (1,3-DPG). Thiurams were by far the most frequent rubber allergens in these patients (16.2% positive reactions, age- and sex-standardized), and the reaction frequency showed a decline from 20.9% in 1997 to 12.8% in 2000. However, this trend was not statistically significant, and was followed by an increase to 16.5% in 2001. All other rubber allergens showed no time trend at all. Although, according to manufacturers' information, the use of dithiocarbamates and MBT derivatives in rubber glove production increased in recent years, these allergens elicited positive reactions in only about 3% of the patients tested, and showed no increasing trend. Thioureas and 1,3-DPG are not widely used in rubber glove production, and play only a minor role in rubber glove contact allergy. Most of the positive reactions to 1,3-DPG are probably false-positive, irritant reactions. PMID- 12641578 TI - Textile dyes sensitization: a study of 49 patients allergic to disperse dye alone. PMID- 12641579 TI - Patch test sensitization to methylchloroisothiazolinone + methylisothiazolinone and 4,4'-diaminodiphenylmethane. PMID- 12641580 TI - Photoaggravated allergic contact dermatitis due to Rosmarinus officinalis cross reactive with Thymus vulgaris. PMID- 12641581 TI - Failure to induce sensitization to budesonide in the guinea pig. PMID- 12641582 TI - Contact allergy to octocrylene. PMID- 12641583 TI - Metal allergy resurfaces in failed hip endoprostheses. PMID- 12641584 TI - Allergic contact dermatitis from isopalmityl diglyceryl sebacate in lipsticks. PMID- 12641585 TI - Contact urticaria photoinduced by benzophenones. PMID- 12641586 TI - Procedure for supplying fragrance information promptly to dermatologists. PMID- 12641587 TI - Early childhood caries in northern Philippines. AB - OBJECTIVES: To determine prevalence and severity of early childhood caries (ECC) among children in northern Philippines. Further, to describe and determine the impact of child-rearing practices and dental visits on caries status. METHODS: Cross-sectional survey of 993 children aged 2-6 years. Caries was diagnosed based on WHO recommendations. Additional behavioral information was obtained from 452 children aged 3-6 years. RESULTS: Caries prevalence and mean dmft (+/-SD) by age were as follows: 2 years = 59% (4.2 +/- 5.3); 3 years = 85% (7.4 +/- 5.5); 4 years = 90% (8.8 +/- 5.6); 5 years = 94% (9.8 +/- 5.5); and 6 years = 92% (10.1 +/- 5.5). Caries rates mirrored those of developing countries with untreated lesions dominating all ages. Mixed breast and bottle feeding was the norm. Almost half were weaned at more than 2 years old. Majority had toothbrushing practices, but mostly without parental assistance. Toothbrushing was initiated at an average age of 2 years. Only a small proportion had a dental visit, mostly for emergency reasons. Among 3-4-year-old children, a significant increase in caries levels were noted for those who started brushing at a later age, had frequent snacks, and had a dental visit for emergency reasons. For children aged 5-6 years, those who went for emergency visits also had significantly more caries. Results indicated an urgent need to (i) increase awareness that ECC is a public health problem in these areas, (ii) advocate use of fluoride as a public health measure, and (iii) increase access to preventive dental services for preschool children. PMID- 12641588 TI - Validity of two oral health-related quality of life measures. AB - OBJECTIVES: To assess the validity of the Oral Impacts on Daily Performance (OIDP) and the short form of the Oral Health Impact Profile (OHIP 14) in the UK. SETTING: Primary care department at a UK dental hospital. SAMPLE: Consecutive patients. METHOD: Cross-sectional comparison of impacts using OIDP and OHIP 14 against clinical findings, Global Oral Health Ratings and pain. RESULTS: A total of 179 patients participated (83.2% response rate). OIDP had weak face validity because it contained contingency questions. Both instruments were developed from the same theoretical model and appeared to have reasonable content validity. In regression analyses, the number of impacts detected by each measure and the total score using OHIP 14 were related to the presence of oral disease and inversely related to age. No suitable transformation could be found to allow regression analysis of OIDP total scores. OHIP 14 correlated more closely with Global Oral Health Ratings but both measures correlated similarly to the experience of pain (0.43 < r < 0.47). The correlation between OHIP and OIDP scores was +0.78. The use of a simple additive method for calculating the total OHIP 14 score did not compromise its validity. CONCLUSION: Both instruments have some validity as measures of Oral Health-Related Quality of Life (OHRQoL) among dental hospital patients. The superior face, criterion and convergent validity and greater amenability to analysis of OHIP 14 render it more suitable for questionnaire based research and for comparing groups. The additive method may be used to calculate the total score for OHIP 14. PMID- 12641589 TI - Dental caries in postwar Bosnia and Herzegovina. AB - OBJECTIVES: To present the prevalence of dental caries in postwar Bosnia and Herzegovina (BH). METHODS: A survey focused on dmft and DMFT indices was carried out in four cantons of the Federation of BH in 1997. The dental health of 6- and 12-year-olds was assessed in random samples (n = 238 and 318, respectively). DMFT index in adult population was assessed in clients coming to dental offices in the same area (35-44-year-olds; n = 401). All the subjects were clinically investigated. RESULTS: During the 1991-95 war, all four cantons were affected by the migration of population. The population per dentist ratio considerably increased in three cantons. In 6-year-olds, the average dmft (+/-SD) was 4.9 +/- 4.0, and 86% of the children were affected with dental caries. The average DMFT of 12-year-olds was 6.2 +/- 3.9. On the average, 94% of the 12-year-olds were affected with dental caries. Mean DMFT of adult clients was 15.1 +/- 7.0. Almost every adult (98%) was affected with dental caries. CONCLUSIONS: The dental status in postwar BH is rather poor and the mean DMFT in all investigated age groups may be considered high. The detrimental effect of 1991-95 war on social conditions and the health care system should be considered in explaining such poor dental health. Despite the limitations, the presented data may be used to assess the treatment needs, assure the visibility for dental issues in BH, and help the west European countries to plan dental services for refugees from BH. PMID- 12641590 TI - Dental clinic attendance in Soweto, South Africa, before and after the introduction of free primary dental health services. AB - OBJECTIVE: To compare patient attendance rates 1 year before and after the introduction of free primary dental health services in Soweto, South Africa. METHODS: Monthly clinic records for clinic attendance, both casual and booked, and number of dental operators were recorded 1 year either side of April 1996. RESULTS: Patient attendance fluctuated by day and season. There was a mean 46% increase in attendance after free primary dental health services were introduced, with more than a sixfold increase in casual attendees (pain, sepsis) than in booked patients (restorative treatment, dentures, orthodontics). CONCLUSIONS: There was a statistically significant increase in casual patient attendance in the year after free primary dental health care was introduced with an increased dental operator load. PMID- 12641591 TI - The shortened dental arch concept--attitudes of dentists in Tanzania. AB - OBJECTIVES: The shortened dental arch concept is a problem-oriented strategy to reduce complex restorative treatment in the posterior regions. The aim of this study was to determine the attitude of Tanzanian dentists about this concept. METHODS: A questionnaire was sent to all 77 dentists treating patients in Tanzania. The questionnaire contained items to assess the dentist's opinions such as dentist's judgement about the oral function of subjects with shortened dental arches, the type of treatment (or no treatment) of subjects with shortened dental arches and the reactions of patients to the proposal to shorten the dental arch. A short description about the problem-oriented approach and the shortened dental arch concept was included. RESULTS: Most of the responding dentists judged that shortened dental arches can provide satisfactory or acceptable chewing function (71%) and dental appearance (79%). Most dentists (89%) indicated that the shortened dental arch concept, as an example of a problem-oriented approach, has a useful place in clinical practice. However, most dentists (89%) inserted free end acrylic partial dentures in these cases. Dentists with experience to apply the concept indicated that most of the patients have no objections after explanation of the proposal to shorten the dental arch. CONCLUSIONS: From this study it is concluded that the shortened dental arch concept is an acceptable strategy for dentists in Tanzania. However, many dentists doubt its implementation in clinical practice. PMID- 12641592 TI - Are there differences in oral health and oral health behavior between individuals with high and low dental fear? AB - Epidemiological studies of the relationship between dental fear, use of dental services, and oral health in different age groups in a common population are scarce. Dental fear and its relationships are usually described in individuals with high dental fear only. The purposes of this study were to describe the prevalence of dental fear in the Norwegian adult population according to age, and to explore differences in oral health, oral hygiene, and visiting habits between individuals with high and low dental fear. For the present study, data from the Trondelag-94 study were used. The prevalence of dental fear in our study population of adults in Trondelag, Norway was 6.6%. There was a tendency for individuals with high dental fear to engage in avoidance behavior more frequently than the low dental fear group. Individuals with high dental fear had a statistically significantly higher number of decayed surfaces (DS), decayed teeth, (DT) and missing teeth (MT) but a statistically significantly lower number of filled surfaces (FS), filled teeth (FT), functional surfaces (FSS), and functional teeth (FST). There were no differences in DMFS and DMFT between the groups of high and low dental fear. Since one of the superior aims of the dental profession is to help a patient to achieve a high number of functional teeth throughout life, consequently detecting and treating dental fear should therefore be an important aspect of dental processionals' work. PMID- 12641593 TI - Comparison between restorations in the permanent dentition produced by hand and rotary instrumentation--survival after 3 years. AB - OBJECTIVES: In the early 1990s, the Ministry of Education in Syria introduced a school oral health programme based on preventive and educational activities only. A restorative component was missing at that time. A few years later, the Atraumatic Restorative Treatment (ART) approach was considered an option to complement the oral health programme but little was known about the longevity of ART restorations in permanent teeth. The null hypothesis which was researched in this trial was: there is no difference between the survival of restorations placed through the ART approach using glass ionomer and those produced through the traditional approach using amalgam (MTA) in permanent dentitions after 3 years. METHODS: Using a parallel group design, 679 grade 2 children, with a mean age of 7.5 years, participated. A total of 369 children were treated through the ART and 310 children through the MTA approach. Eight dentists produced a total of 1118 single- and multiple-surface restorations. RESULTS: The 3-year dropout of the single-surface restorations was 15.3%. Testing the null hypothesis for all single- and multiple-surface restorations revealed a statistically significant difference between the two approaches in favour of the ART approach (P = 0.04). The study showed a 3-year cumulative survival percentage of single-surface ART and MTA restorations of 82.1 (SE = 1.9%) and 76.9% (SE = 2.3%), respectively. The difference was not statistically significant. The number of 3-year surviving multiple-surface restorations was too low for further analyses. The main failure characteristics for both single-surface ART and MTA restorations were restoration missing and gross marginal defect. The 3-year survival percentage of both single surface ART and MTA restorations varied widely among the eight operators resulting in an operator effect (P = 0.01). CONCLUSIONS: The null hypothesis was rejected in favour of the ART approach. It is recommended to select the ART approach to complement the educational and preventive activities of the school oral health programme in Syria. PMID- 12641594 TI - Assessment of manual restorative treatment (MRT) with amalgam in high-caries Filipino children: results after 2 years. AB - BACKGROUND: The atraumatic restorative treatment (ART), using only hand instruments and glass-ionomer cement as adhesive material is recommended for restorative dental treatment in disadvantaged communities lacking electricity and sophisticated dental equipment. Research is required on more durable restorative materials appropriate for populations with high-caries experience. OBJECTIVES: The aim of the study was to evaluate, under field conditions, the applicability and effectiveness of an encapsulated amalgam as restorative material in ART prepared cavities in permanent teeth of children with high-caries rates. As the definition of ART restricts the manual treatment to adhesive materials the approach used is called the manual restorative treatment (MRT). METHODS: Two dentists and two trained healthcare workers, using hand instruments and an encapsulated amalgam that was mixed with a manually driven triturator, placed a total of 934 restorations in the permanent dentition in 466 children. Due to irregular school attendance of Filipino children only 611 restorations could be evaluated by one independent dentist. The average age at reassessment of restorations was 27 +/- 4.5 months. RESULTS: 93.3% of the restorations were acceptable. The failure rate of occlusal surface restorations was 5.6% compared with 13.6% of buccal surface fillings. The survival rate of large occlusal restorations was 95.1% compared with 93.7% for small restorations. The survival of MRT restorations was not influenced by the experience and professional level of the operator. CONCLUSIONS: Amalgam is a suitable MRT material, especially for extensive occlusal lesions in high-caries populations. Studies of longer duration are needed to confirm this finding. PMID- 12641595 TI - Classifying periodontitis among adolescents: implications for epidemiological research. AB - OBJECTIVES: To evaluate the performance of four clinical classification systems proposed for periodontitis in young subjects when applied to epidemiological data on clinical attachment loss. We assess the extent to which the use of different case definition systems may influence the outcome of descriptive and analytical epidemiological studies. METHODS: The data originate in a screening examination for periodontitis carried out among 9162 high school students. Each of four previously published classification systems was applied to the data. The prevalence of cases according to each system was estimated and the association between case status, as defined by each system, and a set of candidate determinant variables was assessed using multivariable logistic regression analyses. RESULTS: The four classification systems yielded rather different prevalence estimates. For localized periodontitis the estimates varied by a factor of 10, and for generalized periodontitis, these varied by a factor of 30. The results of the logistic regression analyses using the different case definitions essentially confirmed the results of a population-based analysis. However, the precision of the estimates decreased with decreasing numbers of cases identified by the classification systems. CONCLUSIONS: From an epidemiological point of view there is little justification for the use of the complicated classification systems. An approach based on the simple definition of a case as a person with clinical attachment loss, e.g. >/=3 mm, is preferable. PMID- 12641596 TI - Psychosocial consequences of dental fear and anxiety. AB - OBJECTIVES: The aim of this study was to examine the negative psychosocial impacts of dental anxiety in a sample of dentally fearful and anxious individuals recruited from the general population. The associations between psychosocial impacts, dental anxiety scale (DAS) scores and other severe fears were explored. METHODS: One hundred and thirty-five subjects who were anxious or fearful about dental treatment were divided into low and high general fear groups based on the number of other severe fears they reported. Negative psychosocial impacts were assessed using a modified form of the scale developed by Kent et al. (1996). This consisted of three dimensions: psychological reactions, social relationships and avoidance/inhibition. Other measures included self-ratings of oral, general and emotional health and scales to assess self-esteem and morale. RESULTS: Overall, 93.1% of subjects reported one or more impacts. Those in the high-fear group had higher psychosocial impact scores than those in the low-fear group (means of 4.19 vs. 2.85; P < 0.05). Differences were most marked with respect to psychological consequences and avoidance/inhibition. The high-fear group had scores indicative of lower self-esteem and lower morale. Forward stepwise linear and logistic regression analyses indicated that both dental anxiety and general fearfulness contributed to these negative outcomes. However, the latter was a more consistent predictor in that it entered six of seven models generated while the former entered only four. CONCLUSION: The study indicated that dental fear and anxiety have pervasive psychosocial consequences, and that these are more marked among subjects with high levels of general fearfulness. It also provided evidence of the validity of a modified form of the psychosocial impact scale developed by Kent et al. (1996). PMID- 12641597 TI - Trends in dental caries in Latin American and Caribbean 5-6- and 11-13-year-old children: a systematic review. AB - OBJECTIVES: To determine trends in dental caries prevalence and severity in Latin America and Caribbean. METHODS: A systematic review was done of studies published between 1970 and 2000 among 5-6- and 11-13-year-old children that used WHO caries diagnostic criteria. RESULTS: Over the last 30 years, there has been a significant decrease in caries severity for children 5-6 years old and also a significant decrease in caries prevalence and severity for children 11-13 years old. The same broad trends were observed when caries prevalence and severity was analyzed for the last 20 and 10 years but these were not statistically significant. CONCLUSIONS: Evidence of a decrease in dental caries in Latin American and Caribbean children has been shown, although the decrease was less prominent in the past few years. PMID- 12641598 TI - The smoking-periodontitis association in developing nations. PMID- 12641599 TI - The single breath transfer factor (Tl,co) and the transfer coefficient (Kco): a window onto the pulmonary microcirculation. AB - The transfer factor, Tl,co (with the transfer coefficient, Kco, also known as the transfer factor per unit alveolar volume, [Tl/Va]), is one of the most useful clinical tests of pulmonary function, the only one which specifically focuses on pulmonary microcirculation. It was originally devised in 1909 as a physiological tool to assess the diffusive capacity of the lung as a gas exchanger. It was subsequently developed as a clinical tool, but cumbersome analytical techniques delayed its introduction into clinical medicine until 1950s. The physiology of the carbon monoxide transfer factor (also called the diffusing capacity Dl,co) is based on the Roughton-Forster equation which partitions Dl,co, a conductance, into membrane (Dm) and red cell (thetaVc) diffusion conductances. Recent work (1987-2001) suggests that 70-80% of the resistance to CO (and O2) diffusion may reside in the red cell fraction. The clinical implication is that Tl,co and Kco are 'windows' onto the pulmonary microcirculation. As regards reference values for clinical use, Tl,co depends on age, height and gender. Kco, which is actually a rate constant, is independent of gender, and is affected principally by age. A schema is presented for the clinical interpretation of Tl,co. As Tl,co is derived from the product of Kco and the accessible alveolar volume (Va), examination of these two components (Kco and Va) will usually suggest a specific pathophysiological mechanism as the explanation for a reduction in Tl,co. PMID- 12641600 TI - Poincare plot indexes of heart rate variability capture dynamic adaptations after haemodialysis in chronic renal failure patients. AB - The hypothesis that the Poincare plot indexes of heart rate variability (HRV) detect dynamic changes after haemodialysis (HD) over the HRV in haemodynamically stable chronic renal failure (CRF) patients was examined. Minor axis (SD1), major axis (SD2) and the SD1/SD2 ratio were compared against standard HRV indexes in time and frequency domain, in a group of healthy subjects and in a group of CRF patients before and after HD. These indexes were estimated from Poincare plots reconstructed with lags of one, two and four heartbeats. The surrogate data analysis technique was applied in order to discern if only random components or linear features of HRV contribute to its dynamics. None of the standard linear HRV indexes changed after HD. The Poincare plot indexes measured from CRF patients were smaller than the ones measured from healthy subjects. In CRF patients the SD1/SD2 ratio decreased after HD, when a lag of four heartbeats was used (0.68 +/- 0.19 before HD versus 0.55 +/- 0.12 after HD, P<0.05). The presence of deterministic components in HRV were confirmed for all measures of the SD1/SD2 ratio. Moreover, a loss of non-linear components after HD was detected by the surrogate analysis over the SD1/SD2 ratio with a lag of four heartbeats. In conclusion, the SD1/SD2 ratio measured at lag of 4 heartbeats capture dynamic changes after HD upon the HRV of CRF patients that are not solely related to linear autocorrelations of HRV. This suggests that the SD1/SD2 ratio reflects non-linear information of HRV. PMID- 12641601 TI - Acute changes in neuromuscular excitability after exhaustive whole body vibration exercise as compared to exhaustion by squatting exercise. AB - The effects of hard squatting exercise with (VbX+) and without (VbX-) vibration on neuromuscular function were tested in 19 healthy young volunteers. Before and after the exercise, three different tests were performed: maximum serial jumping for 30 s, electromyography during isometric knee extension at 70% of the maximum voluntary torque, and the quantitative analysis of the patellar tendon reflex. Between VbX+ and VbX- values, there was no difference found under baseline conditions. Time to exhaustion was significantly shorter in VbX+ than in VbX- (349 +/- 338 s versus 515 +/- 338 s), but blood lactate (5.49 +/- 2.73 mmol l-1 versus 5.00 +/- 2.26 mmol l-1) and subjectively perceived exertion (rate of perceived exertion values 18.1 +/- 1.2 versus 18.6 +/- 1.6) at the termination of exercise indicate comparable levels of fatigue. After the exercise, comparable effects were observed on jump height, ground contact time, and isometric torque. The vastus lateralis mean frequency during isometric torque, however, was higher after VbX+ than after VbX-. Likewise, the tendon reflex amplitude was significantly greater after VbX+ than after VbX- (4.34 +/- 3.63 Nm versus 1.68 +/ 1.32 Nm). It is followed that in exercise unto comparable degrees of exhaustion and muscular fatigue, superimposed 26 Hz vibration appears to elicit an alteration in neuromuscular recruitment patterns, which apparently enhance neuromuscular excitability. Possibly, this effect may be exploited for the design of future training regimes. PMID- 12641602 TI - Beat-to-beat measurement of the finger arterial pressure pulse shape index at rest and during exercise. AB - Arterial pressure waveform can be characterized by the pulse shape index kpulse determined as kpulse = (Pmean - Pdiast) / (Psyst - Pdiast). For brachial artery, the shape index value of 0.33 is usually applied to approximate Pmean from the measured Psyst and Pdiast. Our purpose was to test whether this value can validly be applied to finger vascular beds under different experimental conditions. By using Finapres, we non-invasively estimated the beat-to-beat values of kpulse in the fingers of young healthy persons in supine position at rest and during a 4 min moderate exercise (rhythmical exercise with the quadricep muscles in combination with handgrip compression). To detect intensive peripheral vasoconstrictions, a laser-Doppler probe was attached to the thumb pulp of the same hand. Periods of 30 s without intensive vasoconstriction for rest, different stages of exercise and recovery were involved in the analysis in every subject. The results demonstrated that the group-averaged value of kpulse (median with a 95% confidence interval) in the fingers of 11 healthy volunteers aged from 20 to 24, equalled 0.33 (0.31- 0.34), 0.31 (0.28-0.34), 0.35 (0.33-0.39) and 0.38 (0.34 0.43) for rest, first and second stages of exercise and recovery, respectively. We conclude that in the fingers of young healthy persons in supine position formula Pmean = Pdiast + 1/3 (Psyst - Pdiast) gives an adequate approximation for rest and low intensity exercise (first stage), and slightly underestimates the actual finger mean blood pressure during moderate exercise (second stage) and recovery. PMID- 12641603 TI - Cerebral oxygenation responses to standing in elderly patients with predominantly diastolic dysfunction. AB - Patients with left ventricular dysfunction may have different orthostatic responses of blood pressure (BP) and cerebral oxygenation than healthy elderly subjects. We investigated orthostatic changes in systemic haemodynamic variables and cerebral oxygenation in 21 elderly patients with heart failure New York Heart Association class I-III in stable condition (age 70-83 years) after withdrawal of furosemide and captopril for 2 weeks, and in 18 healthy elderly subjects (age 70 84 years). Frontal cortical concentration changes of oxyhaemoglobin ([O2Hb]) and deoxyhaemoglobin ([HHb]) were continuously measured by near-infrared spectrophotometry and BP changes by Finapres before and during 10 min of standing. Upon standing [O2Hb] reflecting blood flow, changed by -1.2 +/- 0.9 micromol L-1 (mean +/- SEM) in the patients, whereas it decreased by -4.5 +/- 0.6 micromol L-1 (P<0.01) in the healthy subjects after standing (P<0.05 between groups). [HHb] reflecting the sum of cerebral blood flow, arterial oxygen saturation and cerebral oxygen uptake, increased by 1.5 +/- 0.5 micromol L-1 (P<0.05) and 1.7 +/- 0.6 micromol L-1 (P<0.05), respectively. Compared with healthy elderly subjects, elderly patients with left ventricular dysfunction showed smaller orthostatic [O2Hb] decreases (P<0.01), in relation to higher orthostatic BP rises (P<0.05). These findings indicate that BP changes and an altered cardiovascular balance may influence orthostatic cortical haemodynamic responses in elderly subjects. PMID- 12641604 TI - Effect of acute carbon monoxide exposure on heart rate variability in patients with coronary artery disease. AB - Epidemiological studies have shown that air pollution is associated with increased cardiovascular mortality. Although the pathophysiological mechanisms behind this association have remained largely unknown, it has been suggested that changes in cardiac autonomic function may play a role. In this study, we investigated the association between acute carbon monoxide (CO) exposure and cardiac autonomic function as measured by heart rate variability (HRV) in subjects with stable coronary artery disease. Twenty-four hour ambulatory electrocardiographic recording with simultaneous continuous personal CO concentration monitoring was performed in six male patients with angiographically verified coronary artery disease three times with 1-week intervals. Time domain measures of HRV were calculated for 5-min segments before and during the CO exposure periods. For further analysis CO exposures were divided into low (2.7 p.p.m.) CO exposure periods. The mean of maximum CO levels during 61 CO exposure periods was 4.6 p.p.m. (SD 5.0 p.p.m.). High CO exposure was associated with an increase in the square root of the mean of the sum of the squares of differences between adjacent RR intervals (r-MSSD) (P = 0.034). Heart rate remained unchanged during the CO exposure. In conclusion, acute CO exposure which represented most likely exposure derived from traffic seems to modify cardiac autonomic control in patients with stable coronary artery disease. PMID- 12641605 TI - Relationship between breath-synchronous arterial pressure and heart rate variations during orthostatic stress. AB - It has recently been shown that the phase relationship between respiration induced changes in arterial pressure (AP) and heart rate (HR) are different in supine and upright postures. We wanted to further analyse the coupling between respiration, arterial blood pressure and HR in the time domain, and how this coupling was altered during orthostatic stress. Nine healthy subjects were studied. Respiration-induced changes in AP and HR were recorded during frequency- and volume-controlled breathing. This was done during supine rest with and without lower body negative pressure (-50 mmHg) (LBNP). All experiments were performed after beta1-blockade. Responses were averaged breath-by-breath to enhance the time resolution and to eliminate noise. The respiration-induced changes in arterial pulse pressure (PP) were different between control and LBNP: The peak in PP during the respiratory cycle occurred 0.9 +/- 0.8 (mean +/- SD) s before the onset of inspiration during supine control and 0.8 +/- 2.1 s after the onset of inspiration during LBNP (P = 0.03). These changes in the timing of peak PP significantly distorted the cyclic systolic AP and mean AP fluctuations during LBNP. Despite the altered AP response with LBNP, HR fluctuations closely correlated in time with respiration in all conditions, albeit with a significantly reduced amplitude during LBNP (-49%, P = 0.01). The results points to a lack of coupling between AP and HR during paced breathing and thus suggest that respiratory sinus dysrhythmia at least, to a large extent, is independent of the arterial baroreflex. PMID- 12641606 TI - Whole-body impedance recording--a practical method for the diagnosis of sleep apnoea. AB - The aim of this study was to evaluate the utility of whole-body impedance cardiography (ICGWB) in sleep studies, particularly in sleep apnoea detection. A comparison between simultaneous whole night ICGWB and standard polysomnographic recordings were made in 14 patients with a clinical suspicion of obstructive sleep apnoea, a mean age of 46 years (range 30-63 years) and a mean BMI of 29 kg m-2 (25-47). Obstructive apnoeas, central apnoeas and hypopnoeas all caused characteristic patterns in the ICGWB tracing. For an apnoea-hypopnoea index (AHI) > 15 events h-1, the sensitivity of ICGWB was 89% and the specificity 80%. In conclusion, ICGWB signal includes valuable physiological information that can be effectively used for the detection of sleep apnoea episodes. The method seems promising in cases where the multichannel polysomnography is not applicable or when ICGWB is used for haemodynamic monitoring in seriously ill and postoperative patients. PMID- 12641608 TI - Syncope--an unusual presentation of ventricle dysfunction in a patient with Fontan circulation. AB - We report a case of recurrent syncope in association with moderate ventricular dysfunction and mild AV-valve regurgitation in an 18-year-old girl, 4 years after she underwent total cavopulmonary connection surgery. Cardiac catheterization revealed a transpulmonary gradient of 1-2 mmHg. During exercise, a dramatic fall in blood pressure and blood oxygenation was observed, paralleled by an increase in heart rate and central venous pressure. Although a slight increase in pulmonary vascular resistance could not be excluded, the reaction was interpreted in terms of an extremely low transpulmonary gradient in association with ventricular dysfunction. Five months after heart transplantation, the patient has been completely free from syncope. Fontan circulation usually involves a delicate haemodynamic situation which may necessitate haemodynamic re-evaluations, including dynamic measurements of the central venous pressure during exercise. Also a moderate ventricular dysfunction may result in compromised pulmonary circulation which in turn may lead to syncope during exercise as a result of insufficient systemic circulation. PMID- 12641607 TI - The effect of different knee angles on knee volume measured with the Perometer device in uninjured subjects. AB - The Perometer is an optoelectronic device used to calculate limb volume. However, knee angle effect on knee volume during a Perometer measurement has not been reported. This could be a potential confounding factor in assessing knee volume in subjects with impaired knee mobility and where volume measurements are taken over time. To address this issue, bilateral knee volume measurements were recorded from 20 uninjured subjects in seven different knee angles from 0-60 degrees flexion. All 40 knees were not measured in all seven angles because of obstruction of the measurement frame when some knees were in 50 degrees and 60 degrees flexion. Thus, the volume data was separated into three subgroups for analysis: 40 knees at knee flexion angles 0, 10, 20, 30 and 40 degrees; 36 knees at 50 degrees; and 16 knees at 60 degrees. Repeated measures analysis of variance showed a statistically significant difference between the volume measurements in the three subgroups: 0-40 degrees knee flexion [F(4,156) = 35.146, P<0.001], 50 degrees knee flexion (F(5,175) = 56.826, P<0.001] and 60 degrees knee flexion, [F(6,90) = 45.825, P<0.001]. Subsequent paired t-tests showed statistically significant [P<0.001] differences in knee volume for all angle comparisons except for 0 and 10 degrees (absolute volume difference of 1.2 ml; P = 0.772) and 0 and 20 degrees (absolute volume difference of 10.9 ml; P = 0.036) (Bonferroni adjustment applied). The absolute values of the differences that were shown to be statistically significantly different ranged from 12-47 ml for the 0-40 degrees subgroup; 39-85 ml for the 50 degrees subgroup and 35-107 ml for the 60 degrees subgroup. In conclusion, knee angle affects Perometer knee volume measurements and statistically significant differences occur as knee flexion exceeds 20 degrees with greater differences occurring as the knee is more flexed. Hence, knee angle should be reproduced in re-testing and in side-to-side comparisons when evaluating knee volume with the Perometer. PMID- 12641609 TI - Analysis of red cell and platelet morphology using an imaging-combined flow cytometer. AB - Laser-based flow cytometry and aperture-impedance methods are still the dominant technologies used for cell analysis in haematology, but both are limited to areas such as morphological analysis of red cell shape and high-sensitivity detection of platelet agglutinates and aggregates. Flow cytometry alone does not provide precise measurement of red cell volume without chemical pretreatment before detection and aperture-impedance is still considered the gold standard in the field of particle volume analysis. In the present study, an experimental prototype instrument called the imaging-combined flow cytometer (IFC) was evaluated. The IFC is equipped with an imaging device consisting of a pulse laser, lens units and a charge-coupled device (CCD) camera in addition to the flow-cytometric optical set-up. A personal computer was attached to the instrument to handle images derived from the imaging device. Laser illumination was triggered so that the image of an object was captured for each exposure of the CCD camera. Objects in the images were used to calculate size and shape information and to compute fractal texture features by image processing after each measurement. The advantage of the IFC is that it can capture images of selected cells of interest at the same time as flow-cytometric detection. Estimation of red cell volume, discrimination of red cells and platelets, and detection of platelet agglutinates and aggregates were attempted using the IFC in combination with image processing, It was found that image analysis on the IFC could provide a substitutional function for mean corpuscular volume (MCV) estimation and detection of platelet agglutinates and aggregates. The additional information generated by the IFC may be useful in diagnostic haematology. PMID- 12641610 TI - Automated detection of malaria-associated intraleucocytic haemozoin by Cell-Dyn CD4000 depolarization analysis. AB - Laboratory tests for malaria are only performed if there is clinical suspicion of the disease, and a missed diagnosis contributes substantially to morbidity and mortality. Malaria parasites produce haemozoin, which is able to depolarize light and this allows the automated detection of malaria during routine complete blood count analysis (CBC) with some Abbott Cell-Dyn instruments. In this study, we evaluated the Cell-Dyn CD4000 with 831 blood samples submitted for malaria investigations. Samples were categorized as malaria negative (n = 417), convalescent malaria (n = 64) or malaria positive (n = 350) by reference to thin/thick film microscopy, 'rapid test' procedures, polymerase chain reaction analysis and clinical history. With regard to CD4000 depolarization analysis, a malaria positive CD4000 pattern was ascribed to samples that showed one or more abnormal depolarizing purple events, which corresponded to monocytes containing ingested malaria pigment (haemozoin). Positive CD4000 patterns were observed in 11 of 417, 50 of 64 and 281 of 350 of malaria negative, convalescent malaria and malaria positive samples respectively. The specificity and positive predictive values for malaria (active and convalescent) were very high (97.4 and 96.8%, respectively), while sensitivity and negative predictive values were 80.0 and 83.0% respectively. Depolarization analysis was particularly effective for Plasmodium falciparum malaria but there was lower detection sensitivity for White compared with Black African patients. CD4000 90 degrees depolarization vs 0 degrees analysis revealed a proportion of samples with small nonleucocyte associated depolarizing particles. Appearance of such events in the form of a discrete cluster was associated with P. vivax rather than P. falciparum infection. PMID- 12641612 TI - Determination of hemoglobin levels in the finger using near infrared spectroscopy. AB - We previously demonstrated that near-infrared spectroscopy can be used to measure blood flow. The spectrum of blood is dominated by hemoglobin. Therefore, it should be possible to determine the concentration of hemoglobin in tissue using near-infrared transmittance. We attempted to do this in the finger using a unique handheld multiple wavelength spectrophotometer. We took samples from 73 subjects and performed repeat measurements in several subjects. Hemoglobin level was determined at the time of near-infrared measurement. We performed correlation analysis between the hemoglobin values and the absorbance values. There was a strong correlation between hemoglobin levels and the 14 wavelengths (r = 0.738, n = 121, SEE = 1.7). We categorized the patients by hemoglobin level as either normal (12-16 g/dl), mildly anemic (10-12 g/dl), or moderately anemic (<10 g/dl). There were 21 patients in the low hemoglobin group, 29 in the middle range, and 23 in the normal range. The mean hemoglobin levels were 8.4 +/- 0.3 g/dl for the low group, 10.9 +/- 0.1 g/dl for the mildly anemic group, and 13.8 +/- 0.3 g/dl for the group with normal hemoglobin. There was a clear separation of absorbance values among the three groups. The major differences seen were in the midrange of the spectrum. It is encouraging that this first study of hemoglobin measurement yielded data permitting a discrimination of hemoglobin levels. It is hoped that future refinements will lead eventually to a non-invasive technique for the measurement of blood hemoglobin concentration. PMID- 12641611 TI - An improved method for detecting red cells with hemoglobin H inclusions that does not require glass capillary tubes. AB - alpha-Thalassemia trait is the most common inherited abnormality worldwide. Diagnosis of alpha-thalassemia trait can be difficult as there are no abnormalities detectable by hemoglobin electrophoresis or high-performance liquid chromatography. Detection of individuals with alpha-thalassemia trait, particularly the type present in many Asian populations, is important for genetic counseling purposes, because these individuals are at risk for having offspring with hemoglobin Bart's hydrops fetalis, a fatal condition. The best routine diagnostic method to detect individuals with alpha-thalassemia trait is staining reticulocyte-enriched red cell preparations with brilliant cresyl blue to detect hemoglobin H inclusions. Current methods use centrifugation of microhematocrit tubes to enrich for reticulocytes, which presents a laboratory safety hazard. In this report, we describe an alternative technique to enrich for reticulocytes that does not require glass capillary tubes, but is as effective as the capillary tube method for reticulocyte enrichment and detection of cells containing hemoglobin H inclusions. PMID- 12641613 TI - A randomized trial investigating an iron-rich natural mineral water as a prophylaxis against iron deficiency in pregnancy. AB - Spatone Iron-Plus is a naturally occurring mineral water from Trefriw Wells Spa in Conwy County, North Wales, UK. It contains approximately 0.20 mg of iron per millilitre as ferrous sulphate and has been shown to provide iron in a highly bio available form. A 24 ml sachet contains approximately 5 mg of iron. Iron deficiency is common in the obstetric population. However, compliance with traditional iron supplements is poor because of gastrointestinal side-effects. We designed a randomized, double-blind, placebo-controlled trial. A total of 102 low risk antenatal patients, who were noncompliant with routinely prescribed ferrous sulphate tablets, were randomized to receive 48 ml of Spatone water or placebo. The study was conducted between 22 and 28 weeks gestation. Primary outcome measures were compliance, gastrointestinal side-effects and changes in ferritin levels during the trial period. Compliance in the intervention group was 57% compared with 67% in the control group, P = 0.22. Dyspepsia scores, as determined by a recognized and well-validated questionnaire, did not differ between the two groups. During the trial period, mean ferritin levels fell by 24% in the Spatone Iron-Plus group compared with a mean fall of 51% in ferritin levels among the control group, P = 0.016. PMID- 12641615 TI - Significance of HTLV-1 proviral load quantification by real-time PCR as a surrogate marker for HTLV-1-infected cell count. AB - We developed a real-time (RT) PCR quantitative assay to measure the level of the integrated viral genome of HTLV-1 in host peripheral blood-mononuclear cells (PB MNC) from healthy carriers and patients with adult T-cell leukemia (ATL). All of the clinical specimens were serologically and molecularly characterized by enzyme linked immunosorbent assay (ELISA) and Southern blot hybridization (SBH) analyses. The assay system for quantifying the proviral copy level was sensitive, accurate, and reproducible over a wide range of density from 100 to 0.1% with a coefficient of variation (%) of 4.5 to 9.6. The proviral load of the healthy carriers and patients with ATL was 301 +/- 339 copies per 10(4) MNC (3 +/- 3.4%) on average and varied depending on the ATL cell number and the SBH band-status of single or multiple bands. In ATL cases with multiple bands detected by SBH analysis, their ATL cells were shown to harbor multiple copies within one ATL cell, so that the corrected copy number interpolated by the band number in SBH was closely equivalent to the expected ATL cell number in PB, corresponding to the virus-infected cell burden. The proviral load in healthy carriers ranged from 0.1 to 15% of PB-MNC, and, in combination with the fraction (%) of ATL-like flower cells defined by PB smear morphology, enabled carriers to be subgrouped into three categories. This result indicates that the detection of proviral load by(RT) PCR is sufficient and relevant to monitor the infected cell number in the PB and to evaluate the HTLV-1 pathologic status. PMID- 12641614 TI - Urine biochemical markers of early renal dysfunction are associated with iron overload in beta-thalassaemia. AB - Renal dysfunction in thalassemia patients can be attributed to chronic anemia, and iron overload as well as to desferioxamine (DFO) toxicity. We analyzed the urine of 91 well-maintained homozygous beta-thalassemia patients, with no evidence of renal disease, for early evidence of kidney dysfunction by means of electrophoresis and quantitative biochemical tests. Measurement of liver magnetic resonance imaging (MRI) T2 values and serum ferritin concentration was used to estimate iron overload. In 55 of the 91 patients, urine analysis indicated signs of tubular dysfunction. The urine concentration of albumin and beta 2 microglobulin, as well as the activity of N-acetyl-beta-D-glucosaminidase (NAG), correlated positively with serum ferritin concentration and liver iron deposition, as detected by MRI T2 values. This suggested that the cause of renal dysfunction in homozygous beta-thalassemia is iron overload. On the other hand, the same urine markers did not correlate with age, indicating that chronic anemia or desferrioxamine (DFO) treatment are not related to renal dysfunction in thalassemia. PMID- 12641617 TI - A simple and safe nomogram for the management of oral anticoagulation prior to minor surgery. AB - In 80 consecutive, anticoagulated patients scheduled for minor surgery, we reduced warfarin daily dosage by 50% on days 4, 3 and 2 before the surgery, restoring the original dose the day immediately before surgery. The evening after surgery, patients took a double warfarin dose, and then the usual maintenance dose was reintroduced. The mean International Normalized Ratio (INR) value assessed 1 week before surgery was 2.63 (range 1.88-3.87); it decreased at the moment of performing surgery to 1.68 (range 1.42-2.20; P < 0.05 with respect to the preoperative value), and returned to 2.43 7 days after (range 1.96-3.51, P = ns with respect to the preoperative value). No significant difference was found comparing prothrombin fragment 1.2 (F1.2) levels 1 week before surgery and on the morning of surgery (0.49 ng/ml vs 0.67 ng/ml, P = ns), suggesting that no activation of blood coagulation had taken place following the reduction of anticoagulant therapy. Patients developed neither major nor minor bleeding, nor thromboembolism during the procedures or up to 1 month after surgery. In our experience, this method for the management of anticoagulation before minor surgery has been shown to be safe and useful, avoiding the cumbersome shift to either intravenous or subcutaneous heparin. PMID- 12641616 TI - Morphological effects of imatinib mesylate (STI571) on the bone marrow and blood of patients with Philadelphia chromosome (Ph) positive chronic myeloid leukaemia. AB - There have been few reports on the morphological findings in patients with chronic myeloid leukaemia (CML) undergoing treatment with imatinib mesylate. We examined morphological changes in the marrow and peripheral blood of 27 patients with chronic phase (CP), accelerated phase (AP) and blastic phase (BP) CML, 3 and 6 months after treatment with imatinib. At 3 months there was a significant clearance of leukaemic cells as evidenced by a complete haematological response (CHR) in the peripheral blood in 25 patients, together with reduced marrow cellularity in 25 (median reduction CP 42%; AP/BP 68%) and a reduction in the number of megakaryocytes (13 of 18 CP: five of six AP/BP; three patients did not have an assessable marrow) with an increase in the amount of normal megakaryopoiesis. After 6 months, there was continued morphological improvement in eight of 17 CP patients (one patient died after 3 months) with continued cytogenetic response (7 out of 15 patients with assessable metaphases had no abnormal Ph+ cells and three had <35% Ph+ cells) and maintenance of haematologic response in all patients. After an initial response to treatment at 3 months in the AP/BP group, with CHR and a reduction in cellularity in all patients, we found morphological evidence of a loss of response to treatment, with an increase in leukaemic cells, as evidenced by loss of CHR in three of nine and an increase in median cellularity in five patients. No patient in this group achieved a complete cytogenetic response. In summary, in CML-CP patients treated with 6 months of imatinib, there was a significant reduction in leukaemic cells as evidenced by a haematolological response in the peripheral blood, together with reduced marrow cellularity, restoration of morphologically normal haemopoiesis and a meaningful cytogenetic response. Maintaining a response to treatment appeared less likely to occur in the AP/BP group patients, especially those who did not achieve any cytogenetic response to treatment. PMID- 12641618 TI - Open heart surgery with mitral valve replacement--ordeal of an undiagnosed haemophilia patient. PMID- 12641619 TI - Primary medical therapy for acromegaly. AB - There is now considerable evidence that the clinical outcome in patients with acromegaly can be improved very substantially by means of better surgical expertise and effective medical therapies used in a flexible and innovative manner. Medical therapy alone in patients who have not undergone surgery or radiotherapy (primary medical therapy) offers the prospect of near normalisation of GH/IGF-I levels together with substantial tumour shrinkage in a significant number of patients. PMID- 12641620 TI - Guidelines for the management of thyroid cancer. PMID- 12641621 TI - Blocking-type anti-TSH receptor antibodies and relation to responsiveness to antithyroid drug therapy and remission in Graves' disease. AB - OBJECTIVE: Antithyroid drugs are effective in some patients with Graves' disease but not in others. The factors responsible for this difference are still unknown. We examined the relationship between the nature of anti-TSH receptor (TSH-R) antibodies and responsiveness to drugs in Graves' disease. PATIENTS: Twenty-eight untreated patients with Graves' disease were treated with thiamazole and followed for up to 13 years. MEASUREMENT: Antithyroid microsomal antibodies (MCHAs) and antithyroglobulin antibodies (TGHAs) were measured by the passive haemagglutination method. Anti-TSH-R antibodies were measured by a radioreceptor assay (TBII), and thyroid-stimulating antibodies (TSAbs) and TSH-stimulation blocking antibodies (TSBAbs) were measured by bioassays using FRTL-5 cells. Blocking antibodies were also measured by the conversion assay. In order to confirm the usefulness of the conversion assay, in vitro experiments using mixtures of TSAb serum and TSBAb serum were performed. RESULTS: In in vitro conversion experiments, the conversion assay sensitively detected coexisting blocking antibodies. TSBAb was found in seven patients and a positive conversion ratio was found in four patients, and, of these, three patients had both antibodies. Finally, eight patients (28.6%) had blocking antibodies and 25 of 28 patients (89.3%) had stimulating antibodies. These patients with blocking antibodies (Group A) responded well initially to antithyroid drugs and showed earlier normalization of the serum T4 level (3.0 +/- 1.2 weeks) than patients without blocking antibodies (Group B, 10.7 +/- 8.5, P < 0.001). Unexpectedly, remission of Graves' thyrotoxicosis was earlier in Group B (5.1 +/- 4.4 years) than in Group A (8.0 +/- 4.3 years, P < 0.05). Other parameters, including serum T4, goitre size, ophthalmopathy, TBII, TSAb, TGHA and MCHA, were not different between the two groups. CONCLUSIONS: Graves' patients with coexisting blocking antibodies initially respond well to thiamazole but are relatively slow to achieve remission. Measurement of blocking antibodies may be useful for selection of treatment options in Graves' disease. PMID- 12641622 TI - Two novel mutations in the gene encoding thyroxine-binding globulin (TBG) as a cause of complete TBG deficiency in Taiwan. AB - OBJECTIVE: Thyroxine-binding globulin (TBG) encoded by the TBG gene on chromosome Xq22 is the major transport protein, carrying approximately 75% of circulating T4. Inherited defects in TBG are associated with three phenotypes based on the level of TBG in serum of affected hemizygous males: complete TBG deficiency (TBG CD), partial TBG deficiency (TBG-PD) and TBG excess (TBG-E). In this study, we report two unrelated Han Chinese males with complete TBG deficiency who carry different mutations in the TBG gene. PATIENTS: Two index cases of Han males who were diagnosed as having TBG deficiency on the basis of undetectable serum TBG and an additional 75 (50 males and 25 females) normal Han Chinese. MEASUREMENT: Serum thyroid hormones were measured by chemiluminescent immunoassay, thyroid autoantibodies by an agglutination test, and TSH receptor antibody and TBG by radioimmunoassay. Genomic DNA extraction, polymerase chain reaction (PCR) and DNA sequence analysis of the TBG gene were performed with standard methods. RESULTS: One index case had one missense mutation in his copy of the gene, a G --> A transition in codon 52 that results in the replacement of serine by asparagine, and a known polymorphism in codon 283 (TTG --> TTt) that results in the replacement of leucine by phenylalanine. The allelic frequency of TBG-Poly allele in 75 normal Han Chinese (100 chromosomes) was 31%. A second index case was hemizygous for a nonsense mutation in codon 280 of exon 3 (TGG --> TGa). This mutation, located in the C-terminal of TBG, predicts a markedly truncated protein. CONCLUSIONS: This is the first report of complete thyroxine-binding globulin deficiency (TBG-CDT1 and TBG-CDT2) due to TBG gene mutations in Taiwan. PMID- 12641623 TI - Chylomicron remnant metabolism studied with a new breath test in postmenopausal women with and without type 2 diabetes mellitus. AB - OBJECTIVES: The kinetic basis for the effect of type 2 diabetes mellitus (DM) on postprandial lipoproteins has not been fully established. We investigated chylomicron remnant metabolism using a stable isotope breath test and fasting measurements of plasma apolipoprotein (apo) B-48 and apoC-III concentrations in postmenopausal women with and without type 2 DM. PATIENTS: Twenty-four postmenopausal women without DM and 14 postmenopausal women with diet-controlled DM of similar age and body mass index (BMI) were studied in the postabsorptive state. METHODS: The fractional catabolic rate (FCR) of an intravenously injected chylomicron remnant-like emulsion was determined from the appearance of 13CO2 in the breath using isotope-ratio mass spectrometry and multicompartmental modelling. apoB-48, a marker of particle number of intestinal lipoproteins, was determined immunoelectrophoretically. apoC-III was measured by immunoturbidimetric assay. RESULTS: Compared with the nondiabetic women, the women with DM had significantly higher plasma apoB-48 concentration (16.40 +/- 1.18 mg/l vs. 13.0 +/- 0.9 mg/l; mean +/- standard error mean; P = 0.021), higher plasma apoC-III concentration (204.24 +/- 15.18 mg/l vs. 170.74 +/- 10.75 mg/l; P = 0.042) and lower FCR of the chylomicron remnant-like emulsion (0.06 +/- 0.05 pools/h vs. 0.12 +/- 0.02 pools/h; P < 0.001). In the diabetic patients, the FCR of the emulsion was correlated significantly with plasma apoB-48 levels (r = 0.641, P = 0.007) but not with apoC-III levels. CONCLUSIONS: In postmenopausal women, diabetes mellitus appears to decrease the catabolism of chylomicron remnants and result in an accumulation of these particles in plasma. This may chiefly be due to decreased clearance by hepatic receptors related to an effect of insulin resistance. Impairment in the catabolism of chylomicron remnants may contribute to increased risk of atherosclerosis in postmenopausal women with type 2 diabetes mellitus. PMID- 12641625 TI - Effects of low-iodide diet on postsurgical radioiodide ablation therapy in patients with differentiated thyroid carcinoma. AB - OBJECTIVE: Most patients with differentiated thyroid carcinoma (DTC) undergo total thyroidectomy followed by routine radioiodide thyroid remnant ablation. Most centres that routinely perform radioiodide ablation prescribe a low-iodide diet (LID) to increase the radioiodide accumulation in thyroid remnants. The efficacy of an LID on thyroid remnant ablation, however, has never been demonstrated convincingly. DESIGN AND METHODS: In a retrospective study, we studied two groups of DTC patients without distant metastases, who had received either a standard diet or an LID during ablation (LID group, n = 59, and control group, n = 61). Both groups were compared for radioiodide uptake in thyroid remnants during ablation and efficacy parameters of remnant ablation, 6 months after ablation. A subgroup without extrathyroidal tumour growth was analysed separately (stages T1-3, N0). RESULTS: In the total group, the LID during ablation decreased the 24-h urinary iodide excretion to 26.6 micro g compared with 158.8 micro g in controls whereas radioiodide uptake in thyroid remnants was increased by 65% (P < 0.001). Six months after ablation, patients were investigated after thyroid hormone withdrawal. In the total group, no significant effects of the LID during ablation were observed on thyroglobulin (Tg) or the percentage of patients with persistent neck activity after 185 MBq 131I. However, in the LID group, 65% of patients without Tg antibodies had undergone successful ablation (defined by absent neck activity and Tg < 2 micro g/l) compared with 48% in the control group (P < 0.001). In the subgroup (T1-3, N0), 8% of the patients who had undergone the LID had Tg >/= 2 micro g/l vs. 32% in the control group (P = 0.012), whereas successful ablation was achieved in 71% of patients without Tg antibodies in the LID vs. 45% in the control group (P < 0.001). CONCLUSION: We conclude from this study that a low-iodide diet during thyroid remnant ablation improves the efficacy of this treatment. PMID- 12641624 TI - Effects of therapeutic doses of 131I in thyroid papillary carcinoma patients with elevated thyroglobulin level and negative 131I whole-body scan: comparative study. AB - OBJECTIVE: Previous studies have shown a high rate of visualization of uptake and a decrease in serum thyroglobulin (Tg) after therapeutic doses of 131I in well differentiated thyroid cancer patients with elevated thyroglobulinaemia but negative diagnostic 131I whole-body scan (DxWBS), but its therapeutic effect remains controversial. We evaluate the effect of therapeutic doses of 131I in patients with elevated thyroglobulin level but negative DxWBS. DESIGN: Among papillary thyroid carcinoma patients who underwent total or near-total thyroidectomy and remnant ablation with radioiodine during 1996 to 2000 in our hospital, the patients who showed elevated serum Tg levels and no abnormal uptake in DxWBS were selected. The selection for treatment or no treatment was decided according to the preference of the patients, considering side-effects of therapeutic doses of 131I, and the patients were thereafter studied retrospectively. PATIENTS: Sixty papillary thyroid carcinoma patients with elevated thyroglobulinaemia but negative DxWBS were included. Twenty-eight patients were treated, and 32 were untreated. MEASUREMENTS: We compared serum Tg levels measured at less than 3 months before the administration of therapeutic doses of 131I or DxWBS with the levels at 6-12 months after administration between two groups. Comparable data on changes in serum Tg levels during TSH suppression (Tg-on) and those in hypothyroid phase (Tg-off) were available in 25 and 49 patients, respectively. RESULTS: Percentage decreases in both Tg-on and Tg off levels of the treated group [41.2 (10.1-94.1)% and 37.0 (-176.6-88.4)%, respectively] were significantly higher than those of the untreated group [-43.6 (-180.1-7.3)% and -66.6 (-10644.2-39.1)%, respectively] (P < 0.001). The treated patients were followed-up for 23.8 +/- 19.6 months after the administration of therapeutic doses of 131I. In four cases, serum Tg levels converted to negative (< 1.0 ng/ml) both on and off T4 15-22 months after the administration of therapeutic doses of 131I, and negative serum Tg levels persisted for 24-70 months. However, negative conversion of elevated serum Tg levels was not observed in any of the untreated group. Post-treatment WBS revealed pathologic uptake in 12 of 28 cases (42.9%). CONCLUSIONS: This study revealed that the administration of therapeutic doses of 131I has a therapeutic effect, at least for palliation in short-term observation, considering the serum Tg level as an index of tumour burden, and that it can disclose previously undiagnosed lesion in some patients with differentiated thyroid cancer who show elevated thyroglobulin level but negative diagnostic 131I whole-body scan. PMID- 12641626 TI - Evaluation of two health status measures in adults with growth hormone deficiency. AB - OBJECTIVE: To evaluate the psychometric properties of two health status measures for adults with growth hormone deficiency (GHD): Nottingham Health Profile (NHP) and Short-Form Health Survey (SF-36). DESIGN: (1) A cross-sectional survey of adults with treated or untreated GHD to assess reliability and validity of the questionnaires. (2) A randomized, placebo-controlled study of 3 months' GH withdrawal from GH-treated adults to assess the sensitivity of the questionnaires to change. PATIENTS: (1) A cross-sectional survey of 157 patients with severe GHD (peak GH < 10 mU/l on provocative testing), mean age 48.9 years (range 23-70 years), who had either received GH replacement therapy for at least 6 months immediately prior to the study or had not received GH treatment in the previous 6 months. (2) GH treatment was withdrawn from 12 of 21 GH-treated adults, all with severe GHD (peak GH < 7.7 mU/l on provocative testing), mean age 44.9 years (range 25-68 years). MEASUREMENTS: The NHP and SF-36 were used once in the cross sectional survey, but twice in the GH-withdrawal study, at baseline and end-point (after 3 months). RESULTS: (1) Cross-sectional survey. Both questionnaires had high internal consistency reliability with subscale Cronbach's alphas of > 0.73 (NHP) and > 0.78 (SF-36). Calculation of an NHP Total Score, occasionally reported in the literature, was shown to be inadvisable. Overall, patients with GHD were found to have significantly worse perceived functioning than the UK general population in SF-36 subscales of General Health, Bodily Pain, Social Functioning, Physical Functioning, Role-Emotional, Role-Physical, and Vitality. Although neither questionnaire found significant differences between GH-treated and non-GH-treated patients, there were correlations with duration of GH treatment (P < 0.01) for GH-treated patients in SF-36 Mental Health (r = 0.29, N = 87) and SF-36 Vitality (r = 0.33, N = 88), indicating improvement with increasing treatment duration. The SF-36 was also more sensitive than the NHP to sex differences: men had significantly better health status compared with women (P < 0.05) in all SF-36 subscales but Mental Health, but only in one NHP subscale (Physical Mobility). (2) GH-withdrawal study. Significant between-group differences in change were found in SF-36 General Health [t(17) = 2.76, P = 0.013, two-tailed] and SF-36 Mental Health [t(17) = 2.41, P = 0.027, two-tailed]: patients withdrawn from GH reported reduced general health and mental health at end-point. The NHP found no significant change. CONCLUSIONS: The SF-36 is a better measure than the NHP of health status of people with GH deficiency because of its greater discriminatory power, with ability to detect lesser degrees of disability. It also has superior sensitivity to some subgroup differences and superior sensitivity to change compared with the NHP. The SF-36 is highly acceptable to respondents, and has very good internal consistency reliability. The SF-36 is recommended to measure the health status of adults with GH deficiency. PMID- 12641627 TI - A 3-year prospective study to assess uterine growth in girls with Turner's syndrome by pelvic ultrasound. AB - OBJECTIVE: Adult women with Turner's syndrome who have used the donor ovum IVF programme are reported to have reduced pregnancy outcome with an increased risk for first trimester spontaneous abortion. This is considered to be related to a small uterine size and reduced endometrial thickness. This study examines whether adequate oestrogen replacement during the early adolescent years will result in normal adult uterine dimensions, with consequent reduction in these pregnancy risks. DESIGN: A prospective evaluation of uterine dimensions by pelvic ultrasound examination over 3 years, in a group of 18 girls commencing pubertal induction with oestrogen or entering puberty spontaneously. PATIENTS: Girls with Turner's syndrome attending the outpatient clinc at the Royal Children's Hospital and due to start oestrogen treatment were invited to participate in the study. MEASUREMENTS: Data were collected for clinical parameters of age, pubertal staging, menarche, oestrogen dose and karyotype. Ultrasonographic measurements of uterine length, sagittal and transverse width, endometrial cavity and identification of ovaries were also included. RESULTS: The mean age at commencement of the study was 14.6 years, mean age at final evaluation was 17.1 years. Karyotype was 45XO in 6/18, mosaic in 12/18. Spontaneous pubertal onset occurred in 5/18. One of these later required the addition of oestrogen treatment. Pubertal induction with oestrogen was used in 13/18 girls. A total of 15/18 girls have either achieved spontaneous menarche or are using adult doses of oestrogen and progestogen with regular withdrawal bleeds. All 18 girls have achieved a uterine length of 5.8-8.6 cm (mean 7.04 cm) within the normal adult range (5-8 cm). Mean uterine volume was 30.23 cm3. CONCLUSION: The study suggests that adequate oestrogen replacement in early to mid adolescence mimicking spontaneous timing of puberty results in normal uterine growth and adult uterine dimensions. Further follow-up of uterine growth in these girls is warranted. PMID- 12641628 TI - Feasibility of a randomized trial on adjuvant radio-iodine therapy in differentiated thyroid cancer. AB - BACKGROUND: Justification for adjuvant radio-iodine (I-131) therapy in differentiated thyroid cancer (DTC) is purely based on retrospective data. This is true for ablative therapy and even more so for high-dosage adjuvant schedules. Randomized trials on the latter application are considered impossible due to anticipated formidable sample sizes required in a disease with an overall excellent prognosis like DTC. OBJECTIVE: To develop and validate a model that could stratify for risk of recurrence, rather than survival, as is usually done in prognostic indices, and secondly, to use this model to estimate the sample size required for a randomized trial. DESIGN, PATIENTS AND RESULTS: From databases of three large Dutch centres, we identified 342 consecutive patients without known residual DTC after (near-) total thyroidectomy. Using Cox proportional hazards analysis, a model was validated that clearly distinguished risk categories of recurrence using commonly available baseline variables. The model included age, N stage at presentation and T stage in papillary carcinoma. According to this stratification, a subset of patients at substantial risk for relapse (30-40%) was identified. They could be eligible for a trial assessing the impact of high-dose adjuvant I-131 on recurrence rates. Assuming a clinically relevant effect of 30% reduction of relapses, 290 patients would have to be entered in either arm (alpha 0.05, power 80%). CONCLUSION: We conclude that even though a randomized trial on this issue will be difficult to design and conduct, sample size is not the main problem. PMID- 12641629 TI - Re-assessment of growth hormone secretion in young adult patients with childhood onset growth hormone deficiency. AB - OBJECTIVE: Patients with childhood-onset GH deficiency (coGHD) need retesting in late adolescence or young adulthood to verify whether they need to continue GH treatment. For this purpose the Growth Hormone Research Society (GRS) recommends the insulin tolerance test (ITT), or as an alternative the arginine + growth hormone releasing hormone test (ARG + GHRH test) as a diagnostic tool in adolescents and adults. However, there are no standardized cut-off levels based on normal GH secretion for determining GHD vs. GH sufficiency in young adults for the ITT, the ARG + GHRH test or the pyridostigmine + GHRH (PD + GHRH) test, a further new GH stimulation test. PATIENTS AND MEASUREMENTS: We studied 43 patients (28 with organic coGHD, 15 with idiopathic coGHD; 30 males, 13 females; aged 20.4 years, range 16.2-25.4; body mass index 23.5, range 16.3-35.8) using the ARG [0.5 g/kg intravenously (i.v.)] + GHRH (1 micro g/kg i.v.) test, the PD (120 mg orally) + GHRH (1 micro g/kg i.v.) test and the ITT (0.1 IU/kg i.v.) and compared these data with the results of 40 healthy age- and weight-matched volunteers. RESULTS: The GH response in patients was significantly lower than in healthy controls: ARG + GHRH test, 0.8 micro g/l (interquartile range 0.3-2.6) vs. 51.8 micro g/l (32.6-71.2) in controls (P < 0.0001); PD + GHRH test, 0.9 micro g/l (0.3-1.9) vs. 40.4 micro g/l (27.1-54.4) in controls (P < 0.0001); ITT, 0.1 micro g/l (0.0-0.8) vs. 20.3 micro g/l (14.7-31.7) in controls (P < 0.0001). In the ARG + GHRH test we found a diagnostic sensitivity of 100% and a specificity of 97.5% for a cut-off range from 15.1 to 20.3 micro g/l, in the PD + GHRH test a sensitivity of 100% and a specificity of 97% (cut-off range 9.1-13.1 micro g/l) and in the ITT a sensitivity and specificity of 100% each within a cut off range from 2.7 to 8.8 micro g/l. CONCLUSION: There were no marked differences in sensitivity and specificity in young adults among ARG + GHRH test, PD + GHRH test and the ITT in assessing GH secretion. Because of the lack of side-effects, the ARG + GHRH test is the recommended method for re-evaluation of coGHD in young adults when pituitary GHD is suspected. Furthermore, in adult patient groups where organic pituitary coGHD is common, the ITT may be completely replaced by the ARG + GHRH test. Because of the predominance of hypothalamic GHD in childhood, the ITT is commonly performed for the re-evaluation of patients with childhood-onset GHD because of its mechanism of GH stimulation. The present results confirm the high discriminatory capability of the ITT in young adults. PMID- 12641630 TI - GH-secreting pituitary adenomas infrequently contain inactivating mutations of PRKAR1A and LOH of 17q23-24. AB - OBJECTIVE: The molecular events leading to the development of GH-secreting pituitary tumours remain largely unknown. Gsalpha (GNAS1) mutations are found in 27-43% of sporadic GH-secreting adenomas in the Caucasian population, but the frequency of GNAS1 mutations in Japanese and Korean acromegalic patients was reported to be lower, 4-9% and 16%, respectively. Other genes responsible for the tumourigenesis of GH-secreting pituitary adenomas have not been detected yet. PRKAR1A, which codes for the RIalpha regulatory subunit of cyclic AMP-dependent protein kinase A (PKA) on 17q23-24, was recently reported to contain inactivating mutations in some Carney complex families, which involved GH-secreting adenomas in about 10%. We re-evaluated the frequency of GNAS1 mutations and investigated PRKAR1A on the hypothesis that it might play a role in the tumourigenesis of GH secreting adenomas. DESIGN: We analysed exons 8 and 9 of GNAS1 and all exons and the exon-intron boundaries of PRKAR1A with the PCR and by direct sequencing using genomic DNA extracted from 32 GH-secreting pituitary adenomas (30 GH-secreting adenomas, two GH and PRL-secreting adenomas) and 28 corresponding peripheral blood samples, and performed loss of heterozygosity (LOH) analysis of 17q23-24 with four microsatellite markers and intragenic markers of PRKAR1A. RESULTS: Seventeen of 32 (53.1%) tumours showed somatic-activating mutations of GNAS1: 16 (53.3%) of 30 GH-secreting adenomas and one of two GH and PRL-secreting adenomas. Neither inactivating somatic mutations of PRKAR1A nor LOH of 17q23-24 were detected in any of the tumours examined. CONCLUSION: We reconfirm the important role of activating mutations of GNAS1 in GH-secreting adenomas, and conclude that PRKAR1A does not play a significant role in the tumourigenesis. PMID- 12641631 TI - Preoperative lanreotide treatment for GH-secreting pituitary adenomas: effect on tumour volume and predictive factors of significant tumour shrinkage. AB - OBJECTIVE: The objective of this open study of 104 patients was to determine whether the somatostatin analogue lanreotide shrinks GH-secreting adenomas and to identify the predictive factors of a significant tumour volume reduction (> 20%). PATIENTS: A total of 104 previously untreated and newly diagnosed acromegalic patients received the prolonged release (PR) formulation of lanreotide (lanreotide 30 mg, one intramuscular injection every 10 days) for either 1 (n = 84), 2 (n = 13), or 3 or more (n = 7) months before transsphenoidal surgery. MEASUREMENTS: Pituitary tumour volumes, tumour extension grade and possible cavernous sinus invasion were assessed in blinded conditions by a centralized team of radiologists. Factors such as demographics, tumour characteristics, GH and IGF-I levels were evaluated as possible predictive factors of a significant tumour volume reduction. The clinical activity and random GH, IGF-I, IGFBP-3, PRL, TSH, free T4 and lanreotide levels serum concentrations were measured under basal conditions and in the 10 days before surgery. All analyses were done in a centralized laboratory. The tolerability of preoperative PR lanreotide and the surgical outcome at the 6th month after surgery were assessed. RESULTS: The presurgical treatment improved the symptoms of acromegaly and induced a statistically significant reduction of GH, IGF-I and IGFBP-3. Despite the short duration of the preoperative lanreotide 30 mg treatment, IGF-I levels were normalized in 25% of patients. A statistically significant reduction in tumour volume (P < 0.001) was observed. The median value of the differences was -152 mm3. A reduction in tumour volume was observed in 66% of patients and was > 20% in 29% of all patients included. Both the univariate analyses and the logistic regression model demonstrated that a positive hormone response to preoperative lanreotide 30 mg was the sole predictive factor of a significant tumour shrinkage (odds ratio of 7.8, 95% confidence interval 1.6-37.1). Preoperative PR lanreotide did not modify the expected soft consistence of the tumour. The main adverse events consisted of minor gastrointestinal problems. Univariate analyses revealed that younger age, higher GH and IGF-I levels at diagnosis, higher preoperative tumour volume, more than one tumour extrasellar extension and the presence of cavernous sinus invasion were statistically significant determinants of persistent disease at the 6th month after surgery. The multivariate analysis revealed that higher IGF-I levels at diagnosis and the preoperative cavernous sinus invasion were each statistically significant prognostic factors of persistent disease. CONCLUSIONS: A short administration of preoperative lanreotide 30 mg induced a statistically significant shrinkage of GH-secreting pituitary adenomas where this reduction was > 20% of the pretreatment value in 29% of the whole population. Among the factors considered was the fact that positive hormone response to preoperative lanreotide 30 mg was the sole predictive factor of this significant tumour volume reduction. PMID- 12641632 TI - Gender variation in leptin circadian rhythm and pulsatility in adult growth hormone deficiency: effects of growth hormone replacement. AB - OBJECTIVES: Adult growth hormone deficiency (AGHD) is characterized by obesity and associated with increased leptin concentration and decreased leptin pulsatility. Growth hormone replacement (GHR) results in a decrease in leptin concentration and increase in leptin pulsatility, followed by reduction in body fat mass (BFM). In both health and AGHD, women exhibit relatively higher leptin concentrations compared to men. The effect of gender on leptin rhythm and pulse parameters in AGHD is yet to be defined and the gender difference in the response of leptin secretory pattern to GHR has not been determined. Therefore the aim of this study was to evaluate the effect of gender on circadian and pulse parameters of leptin secretion in AGHD, and examine the gender variation in response of these parameters to GHR. STUDY DESIGN: A prospective, open treatment design study to determine the effect of gender on leptin rhythm and pulse parameters in untreated and treated AGHD. GH was commenced at a daily dose of 0.5 IU, and titrated up by increments of 0.25 IU at 2-weekly intervals to achieve and maintain IGF-I SDs between the median and upper end of the age-related reference range. PATIENTS: Twelve patients (six men, six women) with severe AGHD following pituitary surgery, defined as peak GH response < 9 mU/l to provocative testing were studied. All patients required additional pituitary replacement hormones following pituitary surgery and were on optimal doses at recruitment. MEASUREMENTS: Plasma leptin was measured at half-hourly intervals for 24 h, before and 1 month after initiation of GHR. Cosinor analysis was used to determine the circadian rhythm parameters: MESOR (rhythm-adjusted mean), acrophase and amplitude; and ULTRA algorithm used for pulse analysis. Body composition was measured using bioelectrical impedance. RESULTS: BFM was higher in women than men at both visits (P < 0.05), but there was no significant change in BFM in either gender following 1 month of GHR. Women had a higher mean 24-h leptin concentration, MESOR, circadian amplitude and pulse amplitude, both before and after GHR (P < 0.05). Following treatment, mean leptin concentration and MESOR decreased significantly in both men and women (P < 0.05), with no significant difference in percentage change between the genders. Pulse frequency increased and duration decreased significantly after GHR in both groups, without any significant gender difference. IGF-I and IGF SDs were similar in both genders at baseline (P = 0.93). However, after 1 month GHR, the increase in both measurements was greater in men than women (P = 0.005) and men had significantly higher IGF-I and IGF SDs than women (P = 0.01). CONCLUSIONS: As in healthy individuals, leptin levels were higher in women with AGHD than men, both prior to and after GHR. Decline in leptin concentrations and increase in leptin pulsatility following 1 month of GH treatment were similar in both genders. Changes in leptin secretory parameters appeared to occur without any significant decrease in BFM, suggesting a regulatory role for GH. Additionally, the action of GH on leptin secretory pattern does not appear to be mediated by IGF-I. Our data suggest that changes in leptin concentration and rhythm parameters following GHR are independent of gender. PMID- 12641633 TI - Calcitriol regulates the expression of the genes encoding the three key vitamin D3 hydroxylases and the drug-metabolizing enzyme CYP3A4 in the human fetal intestine. AB - BACKGROUND AND AIMS: The human fetal jejunum has been shown to harbour the vitamin D3 (D3) nuclear receptor (VDRn) and to be responsive to calcitriol/1,25 dihydroxyvitamin D3[1,25(OH)2D3] through modulation of proliferation and differentiation processes. The aim of the study was to evaluate the presence as well as the effect of 1,25(OH)2D3 exposure on the expression levels of the three key D3-hydroxylase gene transcripts (25-hydroxylase, CYP27A; 24-hydroxylase, CYP24; 1alpha-hydroxylase, CYP27B1) as well as that of the 1,25(OH)2D3-responsive endobiotic/xenobiotic metabolizing enzyme CYP3A4 (which is also considered a major detoxifiying enzyme) in the human proximal and distal intestine.methods Specimens from normal fetuses ranging from 15 to 20 weeks of gestation were obtained following elective termination of normal pregnancies. Intestinal explants were cultured for a period of 24 h or 48 h with 10-7 m 1,25(OH)2D3. All data were compared to paired-control cultures without 1,25(OH)2D3. Total RNA was extracted and cDNA synthesized by RT-PCR. The cDNA obtained was amplified by radioactive PCR, the signal intensity evaluated by densitometric analyses and expressed in relation to the levels of GAPDH. RESULTS: Data indicate that VDRn, the three D3-hydroxylases as well as CYP3A4 are expressed in all segments of the human fetal small intestine and in the colon. Basal expression levels of VDRn, CYP27A, CYP24 and CYP3A4 were found to be similar in the proximal, median and distal jejunum as well an in the proximal and distal colon. In contrast, basal 1alpha-hydroxylase CYP27B1 expression levels were found to be 65% higher in the colon than in the small intestine (P < 0.02). The 1alpha-hydroxylase was also found to be sensitive to 1,25(OH)2D3 with a 31% decrease in its expression levels within 24 h of 1,25(OH)2D3 exposure to reach a 55% decrease after 48 h of incubation in the presence of the hormone (P < 0.05). Furthermore, the levels of the 25-hydroxylase gene transcript were also decreased by 10% within the first 24 h and by 29% after 48 h of incubation in the presence of 1,25(OH)2D3 (P < 0.003). VDRn expression levels were also found to be reduced following incubation in the presence of 1,25(OH)2D3. In contrast, exposure to 1,25(OH)2D3 contributed to a 4.8 fold increase in the expression of the 24-hydroxylase gene transcript within the first 24 h of exposure (P < 0.03), and to a highly significant induction (24, 22 and 1.5 fold over basal values) of the CYP3A4 gene transcript in 3 of the 4 specimens studies. CONCLUSIONS: Collectively, the data illustrate that at mid gestation 1,25(OH)2D3 is fully active in the modulation of all D3-hydroxylases in the human developing intestine. They also show that the detoxifying enzyme CYP3A4 is not only present along the intestinal tract but is also sensitive to 1,25(OH)2D3, indicating that the hormone may be a key element in intestinal development and in the maintenance of the intestinal mucosa integrity in the basal state and in response to damage-inducing agents. PMID- 12641634 TI - Cortisol and the metabolic syndrome in South Asians. AB - OBJECTIVE: The cardiovascular risk factors which comprise the metabolic syndrome are associated with increased hypothalamic-pituitary-adrenal axis (HPAA) activity in some Caucasian populations. South Asians have high rates of cardiovascular disease and its risk factors. We have investigated the relationships between HPAA activity, adiposity and the metabolic syndrome in a South Asian population. DESIGN: Cross-sectional cohort study. PARTICIPANTS: A total of 509 men and women born at the Holdsworth Memorial Hospital, Mysore, South India between 1934 and 1954 and still living in the area. MEASUREMENTS: Fasting 09.00 h cortisol and corticosteroid-binding globulin. The cohort had previously been investigated for features of the metabolic syndrome. RESULTS: At 09.00 h, cortisol concentration was strongly associated with systolic and diastolic blood pressure (r = 0.25 and r = 0.24, respectively; P < 0.001), fasting glucose concentration (r = 0.26; P < 0.001), insulin resistance (r = 0.20; P < 0.001) and fasting triglyceride concentration (r = 0.17; P < 0.001). In general, higher cortisol concentrations added to the effect of adiposity in increasing cardiovascular risk factors, but there was evidence of an interaction between cortisol and adiposity in determining fasting glucose concentration (P = 0.045) and insulin resistance (P = 0.006). CONCLUSIONS: Associations between 09.00 h cortisol concentration and cardiovascular risk factors in this South Asian cohort were stronger than those previously shown in Caucasian populations, despite similar mean cortisol concentrations, and were amplified by adiposity. This suggests that increased glucocorticoid action may contribute to ethnic differences in the prevalence of the metabolic syndrome, particularly among men and women with a higher body mass index. PMID- 12641635 TI - Demonstration of progesterone receptor-mediated gonadotrophin suppression in the human male. AB - OBJECTIVE: Synthetic gestogens in combination with testosterone have potential as a male hormonal contraceptive, predominantly acting by augmenting suppression of gonadotrophin secretion. Little is known, however, of the effects of gestogens in the male. Gestogens have affinity for both androgen and progesterone receptors but the relative contribution of action at these two receptors in gonadotrophin suppression remains unclear. In this study the effects of progesterone, with no significant androgen-receptor affinity are compared to desogestrel, a synthetic gestogen with relatively low affinity for the androgen receptor, on gonadotrophin secretion in normal men. DESIGN: Subjects received either 50 mg progesterone intramuscularly (i.m.) or 300 micro g desogestrel orally daily for 7 days. Frequent blood sampling over 12 h was undertaken before and after drug administration. GnRH [100 micro g intravenously (i.v.)] was administered 2 h before the end of the frequent sampling period. SUBJECTS: Twenty healthy men were randomly allocated to the two treatment groups. RESULTS: Both progesterone and desogestrel administration resulted in decreases in the concentration of both LH and FSH secretion, as well as testosterone. Analysis of the pulsatile nature of LH secretion indicated that both treatments reduced LH pulse amplitude, and that progesterone reduced LH pulse frequency. Progesterone, but not desogestrel, treatment also reduced the increase in LH secretion in response to GnRH. CONCLUSIONS: The effects of progesterone were at least as marked as those of a maximally effective dose of desogestrel. As progesterone has negligible affinity for the androgen receptor, these results are compatible with the suppressive effects of synthetic 19-norgestogens on gonadotrophin secretion in the male being mediated via the progesterone receptor, with its androgenicity contributing minimally to gonadotrophin suppression. PMID- 12641636 TI - Preoperative plasma concentrations of vascular endothelial growth factor and matrix metalloproteinase 9 are associated with stage progression in papillary thyroid cancer. AB - OBJECTIVE: Vascular endothelial growth factor (VEGF), a potent angiogenic peptide, and matrix metalloproteinase 9 (MMP-9), a proteolytic enzyme, are found to be abundantly expressed in several types of cancer and correlate with tumour progression. In this study, we investigated the relationship between preoperative plasma VEGF, MMP-9 levels and disease stages in papillary thyroid cancer. DESIGN: Plasma samples were consecutively collected from 30 patients with papillary thyroid cancer preoperatively (seven males and 23 females with a mean age of 45 +/- 16 years) and control plasmas obtained from 30 patients with benign goitre (seven males and 23 females with a mean age of 44 +/- 18 years) and 23 healthy persons (four males and 19 females with a mean age 44 +/- 15 years). Plasma VEGF and MMP-9 concentrations were determined by enzyme-linked immunosorbent assay. Cancer progression was staged by the TNM classification of the American Joint Committee on Cancer (AJCC) and the International Union Against Cancer (UICC). RESULTS: In thyroid cancer patients, the plasma VEGF and MMP-9 values were higher than those in controls (51.8 +/- 11.5 ng/l vs. 27.0 +/- 0.8 ng/l; 72.3 +/- 23.3 micro g/l vs. 22.1 +/- 3.0 micro g/l, respectively; P < 0.05), and those in benign goitre (51.8 +/- 11.5 ng/l vs. 39.7 +/- 8.8 ng/l; 72.3 +/- 23.3 micro g/l vs. 23.0 +/- 2.2 micro g/l, respectively; P < 0.05). But, there was no difference of plasma VEGF and MMP-9 between patients with goitre and normal subjects. A comparison of VEGF and MMP-9 levels in patients at each cancer stage and in patients with benign nodule found that plasma VEGF and MMP-9 values in TNM stages III and IV, but not stage I or II, were significantly elevated (P < 0.05). When cancer patients were grouped according to clinopathological features, the plasma VEGF and MMP-9 concentrations were both significantly elevated in patients with large tumour size (P < 0.01), lymph node involvement (P < 0.01), extrathyroidal invasion (P < 0.05), distant metastasis (P < 0.05) or advanced stages (P < 0.01). CONCLUSION: Our study demonstrated that circulating VEGF and MMP-9 levels correlated with progression of papillary thyroid cancer, suggesting plasma VEGF and MMP-9 may serve as preoperative adjuvant markers for assessing disease activity in papillary thyroid cancer. However, they should not be used as a diagnostic tool for differentiating malignant from benign thyroid disorders. PMID- 12641637 TI - Clinical presentation and outcome of pituitary adenomas in teenagers. AB - OBJECTIVE: Pituitary adenomas rarely occur in childhood and adolescence, but their mass effect and endocrine abnormalities can compromise both quality and length of life. In this study we evaluated the symptoms at onset and the long term consequences induced in teenagers by functioning or nonfunctioning pituitary adenomas. DESIGN AND PATIENTS: Clinical, biochemical and neuroradiological data of 44 young patients (12 males and 32 females, aged 16.3 +/- 1.9 years at diagnosis) with pituitary adenomas were evaluated retrospectively at baseline and after therapy. Patients underwent surgery, radiotherapy and/or medical treatment depending on clinical history and endocrine secretion of the tumour. Follow-up ranged from 8 to 252 months (median 55 months). MEASUREMENTS: Baseline and dynamic pituitary function were evaluated in all cases at diagnosis and after treatments. Magnetic resonance imaging (MRI) or computed tomography (CT) scan were performed before therapy and during follow-up. Hormone levels were measured using commercial radioimmunologic or immunoradiometric methods. RESULTS: Pituitary macroadenomas (group 1) or microadenomas (group 2) were found in 61% and 39% of cases, respectively. Overall, 68% were PRL-secreting, 7% GH-secreting, 5% ACTH-secreting and 20% nonfunctioning. The most frequent symptoms at onset were oligoamenorrhoea (62%) and galactorrhoea (59%) in the girls, and headache (58%) in the boys. Pubertal development was delayed in 12/27 (44%) cases with macroadenoma. Growth failure was observed in 4/44 (9%) patients (3 in group 1 and 1 in group 2). At diagnosis, hypopituitarism was detected in 10/27 (37%) patients with macroadenoma. Surgery alone cured 4/18 (22%) and 4/9 (44%) patients in group 1 and group 2, respectively. Adjuvant therapies (second surgery and/or radiotherapy and/or medical treatment) cured the disease in 2/13 (15%) patients with macroadenoma and allowed a persistent normalization in other 4/13 (31%) and 2/4 (50%) cases in group 1 and group 2, respectively. Medical treatment alone cured 2/9 (22%) patients with PRL-secreting macroadenoma and normalized PRL levels in another six (66%) with macroprolactinoma and in 2/7 (28%) patients with microprolactinoma. CONCLUSION: Delay of growth was rarely observed in teenagers with pituitary adenomas. At the onset of the disease, many girls complained of oligoamenorrhoea and galactorrhoea, while headache and delay of pubertal development were the symptoms more frequently referred by boys. Surgery alone was effective in a minority of patients and adjuvant therapies were helpful to obtain the remission of the disease in many cases. In patients with PRL-secreting pituitary adenoma, medical treatment, both as first choice or as adjuvant therapy, normalizes serum PRL levels in 14/27 (52%) cases. PMID- 12641638 TI - Multiple endocrine neoplasia type 2A in Chinese families. PMID- 12641640 TI - Recent advances in the understanding of the repeated bout effect: the protective effect against muscle damage from a single bout of eccentric exercise. AB - The repeated bout effect refers to the adaptation whereby a single bout of eccentric exercise protects against muscle damage from subsequent eccentric bouts. While the mechanism for this adaptation is poorly understood there have been significant recent advances in the understanding of this phenomenon. The purpose of this review is to provide an update on previously proposed theories and address new theories that have been advanced. The potential adaptations have been categorized as neural, mechanical and cellular. There is some evidence to suggest that the repeated bout effect is associated with a shift toward greater recruitment of slow twitch motor units. However, the repeated bout effect has been demonstrated with electrically stimulated contractions, indicating that a peripheral, non-neural adaptation predominates. With respect to mechanical adaptations there is evidence that both dynamic and passive muscle stiffness increase with eccentric training but there are no studies on passive or dynamic stiffness adaptations to a single eccentric bout. The role of the cytoskeleton in regulating dynamic stiffness is a possible area for future research. With respect to cellular adaptations there is evidence of longitudinal addition of sarcomeres and adaptations in the inflammatory response following an initial bout of eccentric exercise. Addition of sarcomeres is thought to reduce sarcomere strain during eccentric contractions thereby avoiding sarcomere disruption. Inflammatory adaptations are thought to limit the proliferation of damage that typically occurs in the days following eccentric exercise. In conclusion, there have been significant advances in the understanding of the repeated bout effect, however, a unified theory explaining the mechanism or mechanisms for this protective adaptation remains elusive. PMID- 12641641 TI - Predictors of 6-minute walk test results in lean, obese and morbidly obese women. AB - The aim of this study was first, to assess the presence of medical conditions that might interfere with walking; second, to assess the differences in walking capacity, perceived exertion and physical complaints between lean, obese and morbidly obese women; and third, to identify anthropometric, physical fitness and physical activity variables that contribute to the variability in the distance achieved during a 6-minute walk test in lean and obese women. A total of 85 overweight and obese females (18-65 years, body mass index (BMI) > or = 27.5 kg m(-2)), 133 morbidly obese females (BMI > or = 35 kg m-2) and 82 age-matched sedentary lean female volunteers (BMI < or = 26 kg m(-2)) were recruited. Patients suffering from severe musculoskeletal and cardiopulmonary disease were excluded from the study. Prior to the test, conditions that might interfere with walking and hours of TV watching were asked for. Physical activity pattern was assessed using the Baecke questionnaire. Weight, height, body composition (bioelectrical impedance method), isokinetic concentric quadriceps strength (Cybex) and peak oxygen uptake (peakVO2_bicycle ergometer) were measured. A 6 minute walk test was performed and heart rate, walking distance, Borg rating scale of perceived exertion (RPE) and physical complaints at the end of the test were recorded. In obese and particularly in morbidly obese women suffering from skin friction, urinary stress incontinence, varicose veins, foot static problems and pain, wearing insoles, suffering from knee pain, low back pain or hip arthritis were significantly more prevalent than in lean women (P < 0.05). Morbidly obese women (BMI > 35 kg m(-2)N = 133) walked significantly slower (5.4 km h(-1)) than obese (5.9 km h(-1)) and lean women (7.2 km h(-1), P < 0.05), were more exerted (RPE 13.3, 12.8 and 12.4, respectively, P < 0.05) and complained more frequently of dyspnea (9.1%, 4.7% and 0% resp., P < 0.05) and musculoskeletal pain (34.9%, 17.7% and 11.4% resp., P < 0.05) at the end of the walk. In a multiple regression analysis, 75% of the variance in walking distance could be explained by BMI, peakVO2, quadriceps muscle strength age, and hours TV watching or sports participation. These data suggest that in contrast with lean women, walking ability of obese women is hampered not only by overweight, reduced aerobic capacity and a sedentary life style, but also by perceived discomfort and pain. Advice or programs aimed at increasing walking for exercise also need to address the conditions that interfere with walking, as well as perceived symptoms and walking difficulties in order to improve participation and compliance. PMID- 12641642 TI - Comparison of Kistler 9861A force platform and Chattecx Balance System for measurement of postural sway: correlation and test-retest reliability. AB - Measurement of postural sway has several potential applications in sports medicine. Traditionally, however, rather complicated equipment has been applied. The purpose of the study was to compare two devices based on two different methods of measuring postural sway: (i) a sophisticated Kistler 9861A force platform (KIS)--which all but requires a laboratory setting--and (ii) Chattecx Balance System (CBS)--which is particularly suited for measurement of sway in the dynamic environment of sports. Measurement of sway was performed in 29 subjects twice at baseline and twice at follow-up 2-4 weeks later. One measurement consisted of four 25-second sequences (eyes open, parallel feet; eyes closed, parallel feet; eyes open, tandem Romberg; eyes closed, tandem Romberg). Factor analysis revealed influence of sight and stance on sway, marked interaction between these two factors and a decrease in sway on retest on the same day. Coefficients of variation were--KIS: 0.13-0.23; CBS: 0.11-0.25. Body height was a covariate for all parameters. Kistler 9861A force platform and CBS were correlated (baseline: r(s) = 0.47; follow-up: r(s) = 0.9). These findings suggest that, when the effects of acclimatization and covariance of body height were taken into account, CBS was as reliable and reproducible as KIS in our laboratory. PMID- 12641644 TI - Effect of short-term and intermittent normobaric hypoxia on endogenous erythropoietin isoforms. AB - The aim of the present study was to evaluate whether the Epo isoforms in blood, induced by short-term and intermittent hypoxia, are different from those at normoxia at sea level and if this could be an impediment to the use of a direct Epo doping test based upon the electric charge of the Epo isoforms. Ten healthy subjects, 9 men and 1 woman, participated in the study. Median age was 22 years (range 20-32). Normobaric hypoxia was administered differently in 3 sub-groups; two groups with 12 h hypoxia and 12 h normoxia up to 10 days: IM 2000 and IM 2700 living in 16.2% and 14.9% O2, corresponding to 2000 and 2700 m above sea level, respectively, and training in normoxia. The third group, C 2700, lived in hypoxia, 14.9% O2 corresponding to 2700 m, continuously for 48 h. The mean serum Epo level increased from 10.9 IUL(-1) (range 8.8-12.5) to 23.5 IUL(-1) (15.6 29.1) after 2 days followed by 19.7 IUL(-1) (16.1-24.1) after 10 days exposure for intermittent hypoxia. The highest values 39.5 IUL(-1) (31.5-50) were obtained for the group exposed for continuous hypoxia for 48 h. The median electrophoretic mobility of the serum Epo isoforms was above the cut-off limit of 670 AMU, previously estimated for discrimination between recombinant and endogenous Epo, in all samples taken before and after exposure to hypoxia. The highest values, mean 730 mAMU (range 703-750) were obtained after 10 days of intermittent hypoxia. CONCLUSION: If the method had been used as a doping test, no false positive results would have been registered for the 15 serum samples from the 10 individuals exposed for hypoxia. Thus, the results indicate that the basic principle for direct detection of recombinant Epo doping, based upon the change in electric charge on Epo, can be used also on individuals having lived in a hypoxic milieu. PMID- 12641643 TI - The effects of proprioceptive or strength training on the neuromuscular function of the ACL reconstructed knee: a randomized clinical trial. AB - OBJECTIVES: The purpose of this study was to determine the effects of a proprioceptive training program (PT) vs. a strength training (ST) program on neuromuscular function after anterior cruciate ligament (ACL) reconstruction. The second purpose was to establish the determinants of functional ability for the operated limb. METHODS: Ten participants with unilateral ACL reconstructions were randomly assigned to one of the following 12-week training protocols: (1) isotonic ST, and (2) PT. The outcome measures were: (1) peak torque time of the hamstring muscles (PeakTT), (2) average concentric and eccentric torques of the quadriceps and hamstring muscles, (3) one-legged single hop for distance (SLHD), (4) one-legged time hop (TH), and (5) subjective scores. RESULTS: : There was a significant group by time interaction effect for PeakTT (P = 0.017). The PT group demonstrated greater percent change in isokinetic torques than the ST group at the end of the 12 weeks (P < or = 0.05). Participants in both groups demonstrated similar significant gains in functional ability and subjective scores (P < or = 0.014). Quadriceps strength is a determinant of functional ability for the operated limb (R2 = 0.72). CONCLUSIONS: : Both training protocols influenced PeakTT. The beneficial effects of ST on PeakTT appear to be load-dependent, while sufficient practice may be crucial in maintaining PeakTT improvements induced by PT. Proprioceptive training alone can induce isokinetic strength gains. Restoring and increasing quadriceps strength is essential to maximize functional ability of the operated knee joint. PMID- 12641645 TI - Secular trends in body dimensions and physical fitness among adolescents in Sweden from 1974 to 1995. AB - AIM: To investigate changes over time in body dimensions, and muscular and aerobic fitness in a representative sample of 16-year-old-girls and boys in secondary schools in Sweden from 1974 to 1995 (n = 855). METHOD: Height and weight were measured and five tests were performed: run-walk, two-hand-lift, Sargent jump, sit-ups, and bench-press. RESULTS: In 1995, girls and boys had higher body mass index (BMI) than in 1974. Girls and boys performed less well in bench-press, sit-ups, and run-walk-tests in 1995. Boys, but not girls, performed better in Sargent jump in 1995 than in 1974. Girls and boys performed better in two-hand-lift in 1995 than in 1974. However, after adjustment for body dimensions, there were no differences in performance in two-hand-lift or run-walk tests between 1974 and 1995. CONCLUSION: There was a decreased aerobic fitness and an increased maximal static strength among adolescents in Sweden between 1974 and 1995. These changes were partly due to increased BMI. However, decreased daily physical activity level cannot be excluded as a contributing factor to the decreased aerobic fitness. The reduced performance in muscular endurance in 1995 was not related to increased BMI. Instead, it is suggested that it is to be related to a specific decrease in hip flexion and arm muscle endurance. PMID- 12641646 TI - Nutritional supplements in Norwegian elite athletes--impact of international ranking and advisors. AB - The aims of this study were to investigate (a) the use of nutritional supplements (NS) (vitamins, minerals, Omega 3, antioxidants, ginseng, amino acids, Creatine and energy supplements) in elite athletes of different international ranking (b) why athletes are using NS, and (c) who recommends the elite athletes to use NS. The total population of elite athletes in Norwegian National Teams (n = 1620, 960 males and 660 females aged 15-39 years) and randomly selected (n = 1681) (916 males and 765 females) controls from the general population, were given a questionnaire including questions about use of nutritional supplements (NS), and from whom athletes had received information about nutrition and recommendations to use NS. The response rate was 76% for male and 92% for female athletes and 75% and 81% for male and female controls, respectively. A similar percentage of female athletes (54%) and controls (52%) reported use of one or more NS, but more male athletes (51%) than male controls (32%) used NS (P < 0.001). However, independent of gender, more athletes as compared to controls used minerals (males 26% vs. 8%; females 42% vs. 20%), amino acids (males 12% vs. 4%; females 3% vs. 0), and Creatine (males 12% vs. 2%; females 3% vs. 0). A lower percentage of NS users were observed in the best female athletes (52%) as compared to female athletes with less experience of international competition (73%) (P < 0.01). In male athletes, NS use was independent of international ranking (49%-53%). The coach was the main advisor for use of NS for both male (58%) and female athletes (52%). For male and female athletes, the main reason for using NS was that they felt it was needed in addition to their daily intake (56% and 67%, respectively). Forty one percent of the male and 37% of the female athletes using NS felt they were well informed about nutrition in general and NS. However, 8% of the NS users did not know whether the NS they used was doping classified or not. IN CONCLUSION: we found that a similar percentage of female elite athletes and controls, but a higher percentage of male elite athletes than controls, reported the use of NS. There was a lower percentage of NS use among the top female athletes, but not the top male athletes as compared to the less successful elite athletes. The coach was the main advisor for NS use both for male and female elite athletes. PMID- 12641648 TI - Genetic heterogeneity of autoimmune disorders in the nonobese diabetic mouse. AB - The nonobese diabetic mouse is highly susceptible not only to diabetes but to several autoimmune diseases, and one might suspect that these are controlled by a shared set of genes. However, based on various gene-segregation experiments, it seems that only a few loci are shared and that each disorder is influenced also by a unique set of genes. PMID- 12641649 TI - Sporothrix schenckii lipid inhibits macrophage phagocytosis: involvement of nitric oxide and tumour necrosis factor-alpha. AB - The role of cell-wall compounds in the immune response to sporotrichosis is unknown. The effect of cell-wall compounds and exoantigen obtained from Sporothrix schenckii in macrophage/fungus interaction was analysed with respect to nitric oxide (NO) and tumour necrosis factor-alpha (TNF-alpha). The lipid compound of the cell wall plays an important role in the pathogenesis of this mycosis and was found to inhibit the phagocytic process and to induce high liberation of NO and TNF-alpha in macrophage cultures in the present study. This is a very interesting result because it is the first report about one compound of the fungus S. schenckii that presents this activity. PMID- 12641650 TI - The formyl peptide N-formyl-methionyl-leucyl-phenylalanine downregulates the expression of FcgammaRs in interferon-gamma-activated monocytes/macrophages in vitro and in vivo. AB - N-Formyl peptides are cleavage products of bacterial and mitochondrial proteins that have pro-inflammatory activities and play an important role in antibacterial host defence. FcgammaRI is a receptor for the Fc portion of immunoglobulin G expressed in monocytes that mediates cytotoxicity and is upregulated by interferon-gamma (IFN-gamma) and interleukin-10 (IL-10). In this report, we demonstrate that N-formyl-methionyl-leucyl-phenylalanine (FMLP) downregulates the expression of FcgammaRI in IFN-gamma-treated monocytes, but not in IL-10-treated monocytes. We determine that supernatants obtained from monocytes treated with IFN-gamma and then exposed to FMLP induce the downregulation of FcgammaRI in naive monocytes. This effect is abrogated by the protease inhibitors phenylmethylsulphonyl fluoride and phosphoramidon, which inhibit serine and metalloproteases, respectively. Supernatants from FMLP-treated neutrophils also induce the downregulation of FcgammaRI, when added to naive monocytes. Similar observations were obtained in vivo in a mouse model of chronic inflammation. In vivo, FMLP also downregulates the expression of FcgammaRs in IFN-gamma-activated macrophages. Our results support the existence of a new mechanism through which FMLP could modulate the activity of monocytes/macrophages during bacterial infections. PMID- 12641652 TI - Cross-reaction of anti-simian immunodeficiency virus envelope protein antibodies with human immunoglobulins. AB - It has been recently established that retroviral envelope proteins (REPs) have structural features similar to those of immunoglobulins (Igs). In this study, we asked whether anti-REP antibodies cross-react with human Igs (hIgs). To this end, murine monoclonal antibodies (mMoAbs) that had been raised against a simian immunodeficiency virus (SIV) envelope protein, SIVMac251gp120, were screened for their ability to react with human monoclonal Igs (HMIgs). We show that two HMIgs, RFSJ2 (a rheumatoid factor) and PAMLN6 (a human anti-hIg V region antibody), but not a number of other HMIgs, could be weakly, but consistently, bound by anti SIVMac251gp120 mMoAbs KK17 and KK46, as judged by indirect enzyme-linked immunosorbent assay and a liquid-phase inhibition immunoassay. Both mMoAbs are specific to amino acid residues in the V3 loop of the SIVMac251gp120. The RFSJ2 Ig heavy-chain V region (VH) is coded in part by a human VH gene, VH3-30.3 and includes the idiotope 7B4 (NKYY), which was previously shown to be present in the gp120 protein of a number of HIV-2 and SIV strains. However, an entirely different VH gene codes the PAMLN6 VH region, opening the possibility that epitope(s) shared between SIVMac251gp120 and hIgs may not be limited to the 7B4 idiotope. PMID- 12641651 TI - DNA injection in combination with electroporation: a novel method for vaccination of farmed ruminants. AB - Injection of plasmid DNA encoding antigens into rodents followed by electroporation improved the immune response when compared with injection without electroporation (Widera et al. J Immunol 2000;164:4635-40; Zucchelli et al. J Virol 2000;74:11598-607; Kadowaki et al. Vaccine 2000;18:2779-88). The present study describes the extension of this technology to farm animals, by injecting plasmid DNA encoding mycobacterial antigens (MPB70, Ag85B and Hsp65) into the muscles of goats and cattle using two different types of electrodes, both allowing DNA delivery at the site of electroporation. The animals were vaccinated under local anaesthesia without any observed immediate or long-term distress or discomfort, or any behavioural signs of muscle damage or pathological changes after the electroporation. DNA-injected and electroporated goats showed increased humoral response after the primary vaccination when compared with nonelectroporated animals. Improved T-cell responses following electroporation were observed in hsp65 DNA-vaccinated cattle. DNA injection with or without electroporation did not compromise the specificity of the tuberculin skin test. In conclusion, a protocol applying in vivo electroporation free of side effects to farmed ruminants was established. In addition, we show that DNA vaccination in combination with electroporation can improve the primary immune responses to the encoded antigens. PMID- 12641653 TI - Pretransplant xenogeneic blood transfusions reduce the humoral response in a mouse-to-rat heart transplantation model. AB - A mouse heart transplanted to a rat is rejected promptly 3 days after transplantation, independent of whether cyclosporin A (CyA) is used as an immunosuppressant or not. Adding a short course of deoxyspergualin (DSG) initially, in addition to continuous CyA treatment, results in long-term graft survival and permits retransplantation during CyA monotherapy. In this paper, we have explored the possibility of substituting the initial heart transplant with blood transfusions. Lymphocyte-enriched blood transfusions combined with CyA and an initial course of DSG proved to lower or eliminate the haemagglutinating antibody titre normally seen in acute vascular xenorejection. The therapy, however, did not prolong the mean survival of the cardiac xenograft, but the same treatment protocol could result in either hyperacute rejection or prolonged survival of up to 11 days. In conclusion, this and earlier studies propose that a humoral unresponsiveness can be induced if the recipient vascular circulation is exposed to a xenoantigen in a mouse-to-rat combination. PMID- 12641654 TI - T-cell activation and the development of an apoptosis-resistant CD45RO+ T-cell population. AB - Growing experimental evidence supports a broadening role for the caspases; not only do they participate in the process of apoptosis but also in the control of the cell cycle and cellular proliferation. The biological role of the caspases in the process of T-cell activation and proliferation is still not defined. In the present study, we propose a potential role, by demonstrating an association of T cell receptor-mediated caspase activity with the development of an apoptosis resistant memory CD45RO+ T-cell population. As previously shown by us, a time dependent induction of caspase activity, in the absence of apoptosis, can be observed in CD3-stimulated human peripheral blood lymphocytes. We here show that a population of CD45RO+ cells, with activated caspase-3 and with resistance to tributyltin-induced apoptosis, develops after 3 days of stimulation. A concomitant expression of the anti-apoptotic protein Bcl-xL accompanied the caspase activity and the development of the apoptosis-resistant phenotype. Finally, upon co-culturing with dexamethasone (DEX), the CD3-induced caspase-3 activity was blocked. During this condition, the expression of the activation marker HLA-DR as well as the cellular proliferative response was strongly suppressed. The development of memory cells with a CD45RO+ phenotype was also blocked. Our data support the hypothesis that caspase-3 activity, observed in CD3 stimulated cells, may be an important component in the proliferation process and, furthermore, might play a role for the development of memory T cells, and DEX inhibits this process. PMID- 12641655 TI - Antigen-specific T-cell downregulation by human dendritic cells following blockade of NF-kappaB. AB - Dendritic cells (DCs) are important for presenting antigen to T cells, especially naive T cells. It has recently been shown that blocking the transcription factor, nuclear factor kappa B (NF-kappaB) in human DCs inhibited the mixed leukocyte reaction. The aim of this study was to investigate the effect of blocking NF kappaB in DCs during presentation of antigen to memory T cells in vitro. Peripheral blood monocytes were differentiated into immature DCs with interleukin 4 (IL-4) and granulocyte-macrophage colony-stimulating factor, and pulsed with an immunogenic tetanus toxoid peptide. Upon maturation, the antigen-pulsed DCs were highly effective in presenting antigen to autologous T cells. However, stimulation with antigen-pulsed DCs overexpressing IotakappaBetaalpha, the endogenous inhibitor of NF-kappaB, led to a significant reduction in T-cell proliferation, and decreased production of interferon-gamma, IL-4 and IL-10, whereas transforming growth factor-beta production was low throughout. There was a significant increase in apoptosis of antigen-specific T cells, even in the presence of IL-2, which was not found in resting T cells. Similar findings were observed using a proteasome inhibitor to block NF-kappaB. The effective downregulation of antigen-specific T-cell responses following blockade of NF kappaB in DCs could be a useful approach for immunomodulating inflammatory T-cell responses. PMID- 12641656 TI - Brown Norway rat ovalbumin-specific immunoglobulin E antibodies increase the human basophil expression of CD63 marker. AB - Anaphylactic shock is an immunoglobulin E (IgE)-dependent hypersensitivity. Biological tests like leucocyte histamine release (LHR) and human basophil activation (HBA), frequently used in human allergy, reflect both the amount of IgE fixed on cells and the cellular reactivity. To assess whether serum-specific IgE from Brown Norway (BN) rats prepared for ovalbumin (OVA)-induced anaphylactic shocks can activate human basophils which has a potential interest in experimental allergy: such a test could rapidly assert an IgE sensitization in laboratory animals genetically T-helper 2 (Th2)-predisposed. Rats (n = 39) were immunized three times (day 0, day 5 and day 21) with OVA injected subcutaneously. One week after the third immunization, a shock was induced with an intravenous (i.v.) bolus of OVA. Sensitization was assessed by passive cutaneous anaphylaxis (PCA) test and dosages of serum IgE antibodies anti-OVA by enzyme-linked immunosorbent assay. Blood basophils were counted before and during the shock. Before the shock induction (at day 21), an LHR test was performed on rat blood, and human basophils were sensitized with rat sera. HBA was demonstrated by the increase in the percentage of cells expressing CD63 antigen membrane, measured by flow cytometry. Twenty-one days after the first subcutaneous (s.c.) immunization, the rat serum induced a significant HBA. HBA was observed neither with the same serum previously heated nor with the serum from nonimmunized rats (NIRs). OVA specific IgEs were significantly increased in immunized rat (IR) serum. The PCA test was negative when the serum was previously heated (56 degrees C). We never observed any circulating basophils, and LHR test was negative. After OVA i.v. administration, all IRs died rapidly. HBA testing strongly suggests a mediation by specific IgE in the increase of CD63 in BN rats. Thus, HBA test seems useful in assessing whether an experimental allergy was induced in animals genetically predisposed to an immune response, Th2-mediated, like BN rat. We also conclude that rat basophil activation does not participate in the histamine release during anaphylactic shock in sensitized BN rats. PMID- 12641657 TI - Expression of transforming growth factor-beta isoforms and their receptors in lepromatous and tuberculoid leprosy. AB - Leprosy is an infectious disease with two polar forms, tuberculoid leprosy (TT) and lepromatous leprosy (LL), that are characterized by strong cell-mediated immunity (CMI) and CMI anergy, respectively. Transforming growth factor-beta (TGF beta) belongs to a family of pleiotropic cytokines (TGF-beta1, TGF-beta2 and TGF beta3) that participate in the control of cell differentiation and proliferation, as well as tissue repair. This cytokine family is unique because it suppresses CMI. In this study, we compared the expression of the three TGF-beta isoforms and their receptors in skin biopsies from LL and TT patients (LL = 20; TT = 20) using immunohistochemistry and automated morphometry. The percentage of cells immunostained for the three TGF-beta isoforms and cells positive for the three TGF-beta receptors in the inflammatory infiltrate located in the papillary dermis, reticular dermis and periadnexal tissue were significantly higher in LL than that in TT, with macrophages being the most common and strongest immunoreactive cells. Some lymphocytes, fibroblasts, keratinocytes and epithelial cells from sweat glands and hair roots were also positive. In situ reverse transcription polymerase chain reaction corroborated the capacity of these cells to synthesize TGF-beta1 and TGF-beta receptor 2. This high expression of TGF-beta isoforms and their receptors could contribute to CMI anergy and other clinical characteristic features of leprosy, like skin atrophy. PMID- 12641658 TI - A correlation between migraine, histamine and immunoglobulin e. AB - Although migraine affects about 15% of population and many studies have been performed to find the mechanism and a successful management, the physiopathology of migraine is still largely unknown. The possibility of an immunoglobulin E (IgE)-mediated allergic mechanism and the role of histamine remain controversial. The aim of the present study was to evaluate serum total IgE and histamine levels in migraine patients and the influence of allergy on them. Seventy patients (18 58 years) with migraine without aura were divided into two groups according to their history of allergy (60% with and 40% without allergy). Serum samples were collected during fasting without allowing any premedication during the two periods of attack and remission. There was a control group containing 45 healthy volunteers. Serum total IgE and histamine levels were measured by enzyme-linked immunosorbent assay and fluorimetric methods, respectively. Mean and standard errors of serum histamine (ng/ml) and total IgE (IU/ml) levels were found in the control group to be 48.16 +/- 2.70 and 38.31 +/- 3.20, in the migraine with allergy group 159.11 +/- 4.60 and 303.30 +/- 42.50 and in the migraine without allergy group 105.01 +/- 8.50 and 79.07 +/- 2.70, respectively. Total IgE levels in migraine with allergy group were found to be significantly (P < 0.0001) above that in the control and another group, suggesting an influence of an IgE-mediated mechanism on migraine. Although the plasma histamine levels, which were significantly elevated (P < 0.0001) in patients with migraine, both during headache and symptom-free periods, when compared with the control group, indicate that there is an increased susceptibility to histamine in allergic conditions, this molecule has also an unrelated role in migraine. The relationship between allergy and migraine can be based, in part, on an IgE-mediated mechanism, and histamine release plays an important role. Thus, the avoidance of allergic conditions in migraine patients may be a simple, helpful way for prophylaxis or their treatment. PMID- 12641659 TI - Cellular profile of cytokine production in a patient with visceral leishmaniasis: gammadelta+ T cells express both type 1 cytokines and interleukin-10. AB - The cytokine profile of CD4+, CD8+ T cells, gammadelta+ T cells and natural killer (NK) cells (CD94+CD3-) was studied in a patient with visceral leishmaniasis (VL). The otherwise healthy, human immunodeficiency virus-negative patient acquired the disease in Tuscany, Italy. Diagnosis was made by demonstration of high concentrations of antibodies against Leishmania antigens in serum. Flow cytometry for the detection of intracellular interferon-gamma (IFN gamma), interleukin (IL)-2, IL-4, IL-6, IL-10, IL-13 and tumour necrosis factor (TNF)-alpha expression in peripheral blood mononuclear cells stimulated with phorbol 12-myristate 13-acetate and ionomycin was performed, followed by treatment with liposomal amphotericin B. CD4+ cells were identified as major cytokine-expressing cells, capable of producing both type 1 and type 2 cytokines. A high frequency of IL-4- and IL-13-expressing CD8+ cells was noted. NK cells and gammadelta+ T cells, thought to be involved in innate host defences against Leishmania, expressed IFN-gamma and TNF-alpha. Ten per cent of gammadelta+ T cells expressed IL-10, predominantly together with IFN-gamma, suggesting additional immune-regulatory roles for this T-cell subset in VL. PMID- 12641660 TI - Immunoglobulin G3 and immunoglobulin M isotype plasma levels are influenced by interleukin-1alpha genotype. AB - The immunoglobulin (Ig) plasma levels are known to be, at least partially, genetically regulated, but all the genes involved are not known. Interleukin-1 (IL-1) is a potent proinflammatory cytokine able to serve as an adjuvant for immune responses. IL-1alpha gene is polymorphic, and at least one of the polymorphisms has been identified in the 5' regulatory region of the promoter, a biallelic base exchange (C-->T) at position -889. We set out to study whether the IL-1alpha genotype might contribute to the genetic component seen in the steady state antibody levels of healthy individuals. Four hundred healthy blood donors (218 males and 182 females) were genotyped, and the plasma levels of IgM, IgG as well as IgG subclasses were measured. An association was found between IgG3 plasma levels and the IL-1alpha genotype; the 1.1 homozygotes had increased IgG3 levels compared with the 1.2 heterozygotes (P < 0.001 in males and P = 0.04 in females, Mann-Whitney U-test). A similar significant association was also found between IgM plasma levels and the IL-1alpha genotype in males, but it was no longer present in females; the 1.1 homozygotes had higher IgM levels than the 2.2 homozygotes (P = 0.03, Mann-Whitney U-test). The data suggest that IL-1alpha mediated signals are critical for IgG3 and IgM responses, which are induced by thymus-independent antigens and are important in activating complement. PMID- 12641679 TI - S(+)-ketamine in paediatric anaesthesia. PMID- 12641680 TI - Children's preoperative anxiety and postoperative behaviour. PMID- 12641681 TI - A paediatric trial comparing midazolam/Syrpalta mixture with premixed midazolam syrup (Roche). AB - BACKGROUND: The bitter taste of midazolam is more acceptable to children when the drug is mixed with fruit juice or syrup. We use a thick grape syrup (Syrpalta), and children are sedated in 10-15 min. A premixed cherry-flavoured midazolam solution (Roche), 2 mg.ml (-1), is currently available. It has been our impression that the premixed midazolam has a slower onset of action. Our aim was to evaluate the effects of the midazolam mixtures (midazolam 0.5 mg.kg (-1), 2 mg.ml (-1)) on children's anxiety, sedation, separation anxiety, mask acceptance, and recovery time. METHODS: Seventy-six healthy children, 1-4 years of age, scheduled for elective placement of ear tubes, were enrolled. The trial was double-blinded and randomized. For premedication, one group received the premixed midazolam, and a second group received the midazolam/Syrpalta mixture. An independent blinded observer evaluated the children, using anxiety and sedation scales at baseline, at 5, 10 and 15 min and at parental separation. Mask acceptance and awakening time were evaluated. RESULTS: Children who received the midazolam/Syrpalta mixture had less anxiety at 15 min (P = 0.046) and at parental separation (P < 0.001) than those who received the premixed midazolam solution. Mask acceptance was not different. CONCLUSIONS: We concluded that the midazolam/Syrpalta mixture has a faster onset of action than the premixed midazolam solution. PMID- 12641682 TI - Effective reduction of anxiety and pain during venous cannulation in children: a comparison of analgesic efficacy conferred by nitrous oxide, EMLA and combination. AB - BACKGROUND: EMLA cream is the current technique of choice to reduce pain during venous cannulation in most paediatric practice. Its use is limited by logistic arrangements and failure to improve cooperation and allay anxiety. Nitrous oxide (N2O) would appear to be an effective alternative. A combination technique may be useful in selected patients. METHODS: One hundred and twenty unpremedicated ASA 1 and 2 day surgery patients, aged 8-15 years were randomized into group 1 (EMLA + air/O2), group 2 (50% N2O/50% O2) and group 3 (EMLA + 50% N2O/50% O2). All patients underwent cannulation on the dorsum of the hand with a 22-G intravenous catheter. Pain behaviour before cannulation was assessed by an observer with Children's Hospital of Eastern Ontario Pain Scale (CHEOPS). Pain during cannulation was evaluated with CHEOPS by an observer and Visual Analogue Scale (VAS) (0-100 mm) by the patient. Satisfaction score (0-100%) for the experience were reported by the patient. Degree of ease of cannulation, time for cannulation were assessed. Heart rate, oxygen saturation were compared before, during and after cannulation. RESULTS: The self-reported VAS for group 3 (10.10 +/- 14.99) was significantly lower than group 1 (26.13 +/- 27.59) and group 2 (18.35 +/- 18.11) (P = 0.003). No significant difference existed between VAS for group 1 and 2. There were also significantly more patients with VAS = 0 in group 3 (23/40) versus group 2 (11/40) versus group 1 (10/40), P = 0.004. The satisfactory score in group 3 (93 +/- 9.96) was significantly higher (P = 0.039) than group 1 (81.13 +/- 24.61) and group 2 (84 +/- 22.02). The increase in CHEOPS from before to during cannulation was significant only in group 1 (P = 0.002). There was no significant difference between frequency of patients with side-effects, ease of cannulation and time taken for cannulation in the three groups. CONCLUSIONS: EMLA and 50% N2O are equally effective for pain reduction while a combination technique provides superior analgesia and satisfaction. N2O has an advantage over EMLA in reduction of pain related behaviour in older children. PMID- 12641683 TI - Target concentration of propofol required to insert the laryngeal mask airway in children. AB - BACKGROUND: The aim of this prospective study was to determine the target concentration (CPRED) curves for laryngeal mask airway (LMA) insertion in children. METHODS: The CPRED of propofol required for insertion of the LMA was determined in 72 unpremedicated children, aged 3-10 years, ASA physical status I. CPRED was reached in 2 min according to a sequential allocation. LMA insertion was then performed by an experienced anaesthetist who was unaware of the concentration. Insertion conditions were noted as satisfactory (jaw relaxed, no coughing, gagging, swallowing or laryngeal spasm, minimal limb movement), excellent (no movement at all) or unacceptable if otherwise. RESULTS: The CPRED50 and CPRED90 were 7.86 microg.ml(-l)[95% confidence interval (CI) 6.46-9.26] and 10.71 microg.ml(-l) (95% CI 9.31-12.12), respectively, in the satisfactory group and 10.86 microg.ml(-l) (95% CI 10.14-11.57) and 13.71 microg.ml(-l) (95% CI 12.99-14.42) in the excellent group when calculations were based on Kataria's pharmacokinetics. These figures were increased 1.5-fold when calculations were based on Marsh's pharmacokinetics. With Schuttler's pharmacokinetics, CPRED50 were 4.62 microg.ml(-l) and 13.66 microg.ml(-l) for satisfactory and excellent groups, respectively. CONCLUSIONS: The target-controlled infusion technique for anaesthesia induction and insertion of the LMA was a safe and effective technique in our study. PMID- 12641684 TI - Effects of halothane and sevoflurane on QT dispersion in paediatric patients. AB - BACKGROUND: The QT dispersion (QTd) of the ECG is an indirect measure of heterogeneity of ventricular repolarization which may contribute to complex ventricular arrhythmias. We compared the effects of halothane and sevoflurane on QTd, and heart-rate corrected QT dispersion (QTcd). METHODS: Fifty ASA physical status I patients, aged 5-15 years, undergoing general anaesthesia were studied. A control ECG recording was printed before induction of anaesthesia. In the halothane group, anaesthesia was induced with halothane 4% in 2 : 1 ratio of air : O2 mixture and in the sevoflurane group with sevoflurane 8% in 2 : 1 ratio of air : O2 mixture. The ECG was recorded 1 and 3 min after induction of anaesthesia, 1 and 3 min after the administration of vecuronium 0.08 m.kg(-1) intravenous and 1 and 3 min after the tracheal intubation. All ECGs were analysed by two cardiologists blinded to the anaesthetic. RESULTS: Although QTd increased in both groups following intubation, this difference was not statistically significant when compared with control values. Following intubation five patients in the halothane group had ventricular arrhythmias of short duration, whereas no arrhythmias were recorded in the sevoflurane group (P = 0.052). Following intubation, QTd (45 +/- 15 ms vs 40 +/- 14 ms) and QTcd (60 +/- 17 ms vs 55 +/- 16 ms) values in the halothane group were significantly greater than the sevoflurane group (P < 0.05). CONCLUSION: Neither sevoflurane nor halothane caused a significant increase in QTd compared with control values before induction. Only QTd following intubation was significantly greater in the halothane group than the sevoflurane group. PMID- 12641685 TI - Ketamine and midazolam is an inappropriate preinduction combination in uncooperative children undergoing brief ambulatory procedures. AB - BACKGROUND: We prospectively studied the effects of intramuscular (i.m.) ketamine alone, or combined with midazolam, on mask acceptance and recovery in young children who were uncooperative during induction of anaesthesia. METHODS: The Institutional Review Board (IRB) approval was obtained to study 80 children, 1-3 years, scheduled for bilateral myringotomies and tube insertion (BMT). Mask induction was attempted in all the children. Those who were uncooperative were randomly assigned to one of the four preinduction treatment groups: group I, ketamine 2 mg.kg(-1); group II, ketamine 2 mg.kg(-1) combined with midazolam 0.1 mg.kg(-1); group III, ketamine 2 mg.kg(-1) with midazolam 0.2 mg.kg(-1); or group IV, ketamine 1 mg.kg(-1) with midazolam 0.2 mg.kg(-1). Anaesthesia was continued with nitrous oxide and halothane by facemask. RESULTS: Children in all treatment groups achieved satisfactory sedation in less than 3 min following the administration of the preinduction drug(s). Compared with patients who received halothane induction (comparison group), the use of ketamine alone did not significantly (P > 0.0167, a Bonferroni corrected significance level) delay recovery and discharge times (18.8 +/- 2.5 and 82.5 +/- 30.7 min vs 12.6 +/- 4.6 and 81.0 +/- 33.8 min, P = 0.030 and P = 0.941, respectively). Patients who received ketamine/midazolam combinations, however, had significantly longer recovery and discharge times vs halothane (32.3 +/- 14.0 and 128.0 +/- 36.6 min, P = 0.001, P = 0.007, respectively). These times were so clinically unacceptable, that the study had to be terminated with only 17 patients receiving study drugs. CONCLUSIONS: It is concluded that ketamine/midazolam combination is not appropriate for preinduction of anaesthesia in paediatric ambulatory patients because of unacceptably prolonged recovery and delayed discharge times. PMID- 12641686 TI - Spread of radioopaque dye in the epidural space in infants. AB - BACKGROUND: This prospective study aimed to assess the extent of spread of dye in the epidural space and whether it would vary in direct proportion to the volume when injecting two volumes of dye. METHODS: Ten infants, aged 2-36 days (mean +/- SD, 13.30 +/- 13.68 days) and weighing 1.8-4.5 kg (mean +/- SD, 2.60 +/- 0.97 days), who were undergoing major thoracoabdominal surgery under epidural and general anaesthesia, were studied. At the end of surgery, two volumes of radioopaque dye (omnipaque) 0.5 ml.kg(-1) and 1 ml.kg(-1) were injected into the epidural space at a rate of 1 ml.2 min(-1). The spread was studied by taking X rays after both injections in the left lateral position. RESULTS: There were 10 different patterns of spread in the 10 cases. Uniformly circumferential and cylindrical spread was seen only in one infant. In the others, there were segregated patches of anterior and posterior spread with or without interspersed patches of circumferential spread. There was variation in the extent, location and the density of spread, filling defects and skipped segments with both volumes. Back leak of dye along the needle track was seen in three cases. Statistically, segments were 9.30 +/- 3.68 for 0.5 ml.kg(-1), for 1 ml.kg(-1) 11.50 +/- 3.03, 3.03, S, P=0.014; circumferential spread for 0.5 ml.kg(-1) 2.70 +/- 2.16, for 1 ml.kg(-1) 5.90 +/- 3.14 3.59, P=0.006; anterior spread for 0.5 ml.kg(-1) 3.60 +/- 1.58, for 1 ml kg(-1) 7.90 +/- 2.33 5.88, P=0.001; posterior spread for 0.5 ml.kg(-1) 8.20 +/- 3.71, for 1 ml.kg(-1) 9.80 +/- 3.68 3.54, P=0.006. Doubling of spread with doubling of the volume occurred in only one patient. There was a variable increase in extent or in the density of spread with reduction of skipped segments with the 1 ml.kg-1. The probable reasons for this variable spread and the mechanism of epidural anaesthesia in the presence of such spread are discussed. CONCLUSIONS: There is a difference in quantitative as well as qualitative spread in different patients and in the same patient with different volumes. There were statistically significant increases in the number of segments, circumferential, anterior and posterior locations in the 1.0 ml group. Both extent and density of spread improve with the higher volume but not in direct proportion to volume. 1 ml.kg(-1) has a better quantitative as well as qualitative spread than 0.5 ml and has a better chance of producing adequate anaesthesia. PMID- 12641687 TI - Caudal bupivacaine and s(+)-ketamine for postoperative analgesia in children. AB - BACKGROUND: The aim of this study was to evaluate whether caudal s-ketamine would prolong analgesia together with caudal bupivacaine. METHODS: Thirty children, ASA I-II, 1 month-9 years, scheduled for minor inguinal or penile surgery, were randomly assigned in a prospective, double-blind fashion to receive single shot caudal blockade by either bupivacaine 0.125% 1 ml.kg(-1) (group B, n = 15) or a mixture of bupivacaine 0.125% 1 ml.kg(-1) and preservative-free s-ketamine 0.5 mg.kg(-1) (group K, n = 15). Postoperative pain was assessed by means of an observational 10-point scoring system and analgesia was administered if the pain score exceeded a value of 3. RESULTS: Within 24 h after caudal block 10 patients (67%) in group K and three patients in group B (20%) did not require additional analgesic medication (P < 0.01). There were no significant differences between the groups for incidence of haemodynamic changes. We did not observe any negative side-effects attributable to the caudal block or s-ketamine. CONCLUSIONS: Addition of preservative-free s-ketamine 0.5 mg.kg(-1) to caudal bupivacaine 0.125% 1 ml.kg(-1) provides significant prolongation of analgesia without producing negative side-effects. PMID- 12641688 TI - Tramadol for pain relief in children undergoing tonsillectomy: a comparison with morphine. AB - BACKGROUND: Pain control for paediatric patients undergoing tonsillectomy remains problematic. Tramadol is reported to be an effective analgesic and to have a side effect profile similar to morphine, but is currently not licensed for paediatric use in the UK. METHODS: We conducted a prospective, double-blind, randomized controlled trial in children who were scheduled for elective tonsillectomy or adenotonsillectomy at the Royal Aberdeen Children Hospital. Following local ethics committee approval and after obtaining a drug exemption certificate from the Medicines Licensing Agency for an unlicensed drug, we recruited 20 patients each into morphine (0.1 mg.kg(-1)), tramadol (1 mg.kg(-1)) and tramadol (2 mg.kg( 1)) groups. These drugs were given as a single injection following induction of anaesthesia. In addition, all patients received diclofenac (1 mg.kg(-1)) rectally. The postoperative pain scores, analgesic requirements, sedation scores, signs of respiratory depression and nausea and vomiting, as well as antiemetic requirements, were noted at 4-h intervals until discharge. RESULTS: There were no statistically significant differences in age, weight, type of operation or induction of anaesthesia, 4-h sedation and pain scores and further analgesic requirements. There were no episodes of respiratory depression. Morphine was associated with a significantly higher incidence of vomiting following discharge to the wards (75% versus 40%, P=0.03) compared with both tramadol groups. CONCLUSIONS: Tramadol has similar analgesic properties, when compared with morphine. The various pharmaceutical presentations and the availability as a noncontrolled substance may make it a useful addition to paediatric anaesthesia if it becomes licensed for paediatric anaesthesia in the UK. PMID- 12641689 TI - Spinal anaesthesia in a child with hypoplastic left heart syndrome. AB - Hypoplastic left heart syndrome (HLHS), a complex congenital heart disease, is the most common lethal cardiac defect in neonates. Its treatment includes cardiac transplantation and/or surgical palliation. Associated extracardiac congenital abnormalities are exceptional. We report the case of a neonate with HLHS and anorectal atresia who required urgent surgical management to relieve intestinal obstruction. The surgery was successfully performed under spinal anaesthesia. PMID- 12641690 TI - Anaesthetic implications of Costello syndrome. AB - Costello syndrome is a disorder that primarily involves ectodermal tissues and is characterized by mental and growth retardation, distinctive coarse facies, redundant skin (neck, palms and soles), and papillomata (perioral, nasal and anal). Of primary concern to anaesthesiologists are potential airway difficulties related a short neck, macroglossia, hypertrophied tonsillar and supraglottic tissues, laryngeal papillomata and choanal atresia. A significant percentage of patients also have cardiac involvement which may manifest as congenital heart defects, arrhythmias, valvular dysfunction, or hypertrophic cardiomyopathy. Central nervous system involvement includes developmental delay and seizure disorders while endocrine abnormalities have been reported including hypothalamic pituitary dysfunction resulting in hypopituitarism, hypothyroidism, and hypoadrenalism. The authors present a 2-year old child with Costello syndrome who required anaesthesia for direct laryngoscopy, rigid bronchoscopy, bilateral pressure equalization tubes and tonsillectomy. The perioperative implications of the syndrome are discussed. PMID- 12641691 TI - Anaesthetic and airway management in a child with Hanhart's syndrome. AB - Hanhart's syndrome (oromandibular-limb hypogenesis syndrome) is a rare disease characterized by hypoglossia/aglossia, various distal limb defects and micrognathia. Difficult airway due to micrognathia may complicate anaesthetic management in this syndrome. We describe the anaesthetic management of a child with Hanhart's syndrome undergoing plastic reconstructive surgery. PMID- 12641692 TI - Anaesthetic management for Edward's syndrome. AB - We report the anaesthetic management for a Cohen procedure in a girl with Edward's syndrome (trisomy 18). Information in the literature about anaesthetic management in trisomy 18 patients is poor. The patients are dysmorphic, have severe mental retardation and multiple organ malformations of which congenital heart disease is the most important. Our patient was a 7 year-old girl, such a survival being exceptional with trisomy 18. Anaesthesia is high risk and pain assessment is difficult. Establishing good relations with the family and appraising the need for surgical procedures are the main other difficulties. PMID- 12641693 TI - Anaesthetic management in a case of a type IV laryngotracheo-oesophageal cleft. AB - We report the case of a newborn baby with a type IV laryngotracheo-oesophageal cleft and the anaesthetic management during the rigid bronchoscopy that was performed at 5 days of age. After anaesthetic induction with sevoflurane and atropine, the child was maintained with sevoflurane 2-2.5% and remifentanil at an infusion rate of 0.5 microg.kg(-1).min(-1). Ventilation was managed through the lateral port of the bronchoscope. The patient breathed sevoflurane and oxygen/N2O spontaneously via a Jackson-Rees circuit. To prevent the stomach from filling up with anaesthetic gases, a Foley catheter was placed orally into the stomach. The Foley balloon was inflated and retracted until it sealed the gastro-oesophageal junction. Tracheal intubation was performed after bronchoscopy to allow suture of the stomach into two chambers. Oxygenation was adequate with no air leakage, with spontaneous ventilation. The Foley catheter was removed afterwards and the patient awakened. We review the literature on different ways of managing the airway in these cases and protecting it from gastric aspiration during ventilation. PMID- 12641694 TI - Combined anaesthesia in a young patient with dystrophic epidermolysis bullosa. PMID- 12641695 TI - Anaesthetic management of a child with Hallermann-Streiff Francois syndrome. PMID- 12641696 TI - Hazard when using a cuff pressure regulator with a three-way stopcock for tracheal cuff pressure control. PMID- 12641697 TI - Evolution of accommodative function and development of ocular movements in children. AB - The evolution of the accommodative function and development of ocular movement are evaluated in a non-clinical paediatric population (1056 subjects) aged 6-12 years, providing means for each age in the optometric tests that evaluate the accommodative amplitude, accommodative facility, accommodative response (lag), and saccadic movements. A comparison of these values between ages (anova) established three distinct trends in the behaviour of these parameters. The accommodative amplitude, measured by modified dynamic retinoscopy, and the evaluation of the saccadic movements by the development of ocular movements [developmental eye movement (DEM)] test showed continuous change with age. The values for monocular and binocular accommodative facility, measured by +/-2.00 D flippers, indicated the need to divide the population into two age groups (6-7 and 8-12 years). Finally, the means of accommodative response, measured by monocular estimation model (MEM) retinoscopy, and the direct observation of saccadic movement revealed no significant differences between ages, establishing a single mean reference value for the age group studied. PMID- 12641698 TI - Polarization properties of the in vitro old human crystalline lens. AB - We have studied the spatially resolved polarization properties of the in vitro intact old human crystalline lens (from 56 to 88 years old) by using Mueller matrix imaging polarimetry. Analysis was performed within an average of 54 h of death. Results show that the overall retardation is small (7 degrees on average) and decreases from the centre of the lens to the periphery. Lenticular birefringence is linear but has a spatial dependence, reducing outwards along the radius. The distribution of azimuthal angle of the birefringent structure of the crystalline lens changes depending on each individual lens. Diattenuation and polarizance were found to be small, however, depolarization was about 35% for the set of lenses studied here. PMID- 12641699 TI - Experience and training as determinants of grading reliability when assessing the severity of contact lens complications. AB - PURPOSE: A study was conducted to investigate the influence of experience and training on grading reliability when assessing the severity of contact lens complications. METHODS: Twenty-three optometry students who were unfamiliar with the use of grading scales each used the Efron Grading Tutor computer program to ascertain grading reliability at an 'initial' experimental session and a 'final' session 3 weeks later. Twelve subjects (the 'trained' group) were given a tutorial on grading techniques and were asked to complete two grading exercises between the initial and final sessions. The other 11 subjects (the 'untrained' group) received no such training between the two sessions. Differences in grading reliability between the initial and final grading sessions were evaluated. RESULTS: Grading reliability was superior (lower) for the combined subject cohort at the final session (mean +/- standard deviation 0.33 +/- 0.12) compared with the initial session (0.46 +/- 0.25) (p = 0.004). However, there was no difference in the improvement in grading reliability between the two groups. CONCLUSIONS: Grading reliability improves statistically with some experience, although perhaps not to a clinically meaningful extent. No added benefit can be derived from supplemental training. PMID- 12641700 TI - The UWCDot colour vision test and low vision. AB - PURPOSE: Previous studies have shown that colour vision defects are common in the low vision population even when properly designed tests are used. However, there are very few clinical tests available that are suitable for this group of patients. One of the more common is the Jumbo D-15 (JD15). Although this test uses caps more suitable for the reduced acuity, it requires the patient to have some knowledge of colour order and sufficient dexterity to manipulate the caps. We compared the JD15 with the University of Waterloo Colored Dot test (UWCDot), which has neither of these requirements, to determine whether the UWCDot test could be used as a substitute for the JD15. METHODS: The colour vision of 40 consecutive low vision patients was evaluated with both tests. Acuities ranged from 6/6 to 6/1600 with a median value of 6/30. RESULTS: All subjects could perform both tests. The kappa coefficient of agreement between tests was high at 0.85 when any major crossing was a failure on the JD15 and any mistake was a failure on the UWCDot. Classification of the type of defect was also reasonable when the defect was relatively severe. CONCLUSIONS: The UWCDot can be used as a substitute for the JD15 in the low vision clinic. Both tests identify individuals with moderate-to-severe deficiencies, but the UWCDot does not require any manual dexterity and it does not require knowledge of colour-order. PMID- 12641701 TI - Variation of multifocal electroretinogram with axial length. AB - INTRODUCTION: The first-order kernel response of multifocal electroretinogram (mfERG) decreases in myopia. A recent study indicates that the flash ERG is also reduced with increased axial length. The aim of this study was to investigate the variations in the first-order response (K1) and the first slice of second-order response (K2.1) across the retina for different axial lengths. METHODS: Thirty healthy subjects with axial length from 23.72 to 28.13 mm (spherical equivalent refractive errors from plano to -10.50 D) were recruited for mfERG measurement using VERIS 4.0. All subjects were fully corrected after cycloplegic refraction and pupils were dilated prior to mfERG recording. There is one trough, n1, and one peak, p1, in the K1 response and three troughs, n1, n2, n3, and three peaks, p1, p2, p3, in the K2.1 response. The amplitudes and implicit times of K1 and K2.1 responses were analysed to determine the characteristic of the responses across retina and the correlation to axial length. RESULTS: The amplitudes of p1 (in the first-order kernel-K1) decreased in the central region and the paracentral region (ring 3) as the axial length increased. The central retinal region showed high rates of reduction in both n1 and p1 (in K1). The amplitudes of n1p1 and n2p2 (in the first slice of the second-order kernel-K2.1) were reduced in the paracentral region (from ring 2 to ring 5) as axial length increased. The average n1 and p1 in K1, and n1p1 and n2p2 in K2.1 mfERG responses are decreased in amplitude by 6-10% per millimetre elongation of axial length. CONCLUSION: Eyes with longer axial lengths, usually with high myopia, have a weaker mfERG response and this attenuation is across the measured retina (from central to paracentral regions) but different kernel responses show a different pattern of attenuation at different retinal eccentricities. The weaker mfERG responses may be related to the morphological changes associated with increased axial length. PMID- 12641702 TI - The asphericity, curvature and tilt of the human cornea measured using a videokeratoscope. AB - The EyeSys videokeratoscope (VK) measurements of the principal corneal meridians of 98 subjects already analysed by Douthwaite et al. [Ophthal. Physiol. Opt. (1999)19:467-474] were re-analysed in order to revise the assessment of asphericity, to derive information on corneal tilt and to assess the degree to which the corneal section approximates to that of a conic section. The range of normality for the revised p-value (asphericity) was from 0.57 to 0.97 for the near horizontal and from 0.56 to 1.08 in the near vertical principal meridians. The approximate corneal tilt angles ranged from -3.95 to +8.13 degrees in the horizontal and from -8.99 to +9.33 degrees in the vertical meridian. A tilted conicoidal surface will display a linear relationship (r = 1) when a scatterplot is drawn of the perpendicular distance squared vs radius squared, after first averaging the two semimeridian results for each VK ring. Analysing the results from the human cornea in the same way allows an assessment of the degree to which the corneal section approximates to that of the conic section. PMID- 12641703 TI - Legal issues in contact lens practice with special reference to the practice of orthokeratology. AB - This report aims to encourage optometrists to reflect on the legal implications of clinical contact lens practice, with particular reference to the practice of orthokeratology (ortho-k), which has seen a recent revival of interest. A patient may claim compensation if an optometrist is negligent by breaching his duty to exercise reasonable care and skill in diagnosis, advice or treatment. However, the optometrist will only be liable for reasonably foreseeable harm to the patient, so practitioners need to be fully aware of the foreseeable risks. Failure to adequately inform the patient may lead to a claim for negligence, if disclosure of the risks would have influenced the patient's decision to undergo or forego the procedure. It is important that the professional bodies establish guidelines on acceptable practice, with particular emphasis on the provision of information to patients. Reasonable disclosure, use of appropriate information/consent forms, and proper documentation of cases, should all focus on the needs of, and benefits to, patients. PMID- 12641704 TI - Morphology of corneal nerves in soft contact lens wear. A comparative study using confocal microscopy. AB - The aim of this study was to evaluate corneal innervation in soft contact lens wearers using the Tomey Confoscan confocal microscope (40x/0.75 objective lens). Three distinct age- and sex-matched subject groups were involved, including extended soft (hydrogel and silicone-hydrogel) contact lens wearers, overnight soft (hydrogel) contact lens wearers, and non contact lens wearers. A number of variables were objectively measured, subjectively evaluated, or graded in order to investigate the distribution and morphology of corneal nerves. For most of the evaluated parameters, no statistically significant differences were found. However, qualitative observations showed noticeable differences in corneal nerve appearance among the different subject groups; the degree of corneal oedema was suggested as the main causative factor. In conclusion, neither the short-term (overnight wear) nor the long-term (12-month extended wear) soft contact lens wear appeared to affect the morphology and/or distribution of corneal nerves as viewed with confocal microscopy. PMID- 12641706 TI - Age, lens transmittance, and the possible effects of light on melatonin suppression. AB - Recently it has been suggested that a previously undetected, rhodopsin-based, visual pigment, located in some retinal ganglion cells and having a peak sensitivity around 460 nm, may be responsible for light-induced melatonin suppression and, perhaps, maintenance of the circadian rhythm. Using data from the literature, it is shown that, as absorption in the crystalline lens for shorter visible wavelengths increases substantially with age, while the pupil diameter tends to decrease, the effective retinal exposure received under the same ambient lighting conditions by the pigment is almost 10 times lower in an old, as compared with a young, eye. Interestingly, replacement of the old crystalline lens by an intraocular implant restores the exposure to youthful levels. The possible effects of these changes with age on circadian rhythms are discussed. PMID- 12641705 TI - The projected increase in glaucoma due to an ageing population. AB - The number of cases of primary open angle glaucoma (POAG) in England and Wales is estimated to increase by a third over the 20 years to 2021, and then continue upwards at a similar pace to 2031. The estimates are based on official population projections, organised by 5-year age groups, together with an age-specific prevalence curve for POAG derived from the results of 11 epidemiological prevalence surveys. To cope with additional pressures, a thorough reappraisal of the present system for detection, referral, diagnosis, treatment and monitoring of the disease is likely to be required. PMID- 12641709 TI - Enhanced secretion and low temperature stabilization of a hyperthermostable and Ca2+-independent alpha-amylase of Geobacillus thermoleovorans by surfactants. AB - AIMS: Selection of suitable surfactants for enhancing and stabilizing alpha amylase of Geobacillus thermoleovorans. METHODS AND RESULTS: Geobacillus thermoleovorans was cultivated in shake flasks containing 50 ml of starch-yeast extract-tryptone (SYT) medium with/without surfactants. Titres of the enzyme in media were monitored. The enzyme was also preserved at 4 degrees C with/without surfactants and enzyme activities were determined. Among polyethylene glycol (PEGs) of different molecular weights, PEG 8000 (0.5%, w/v) caused a slight increase in the enzyme titre, while Tween-20, Tween-40 and Tween-60 (0.03%, w/v) exerted a significant stimulatory effect on enzyme secretion. In the presence of SDS, Tween-80 and cholic acid (0.03%, w/v), the enzyme production was nearly twofold higher than that in the control. The anionic (SDS, cholic acid) and non ionic (Tweens) detergents increased the cell membrane permeability, and thus, enhanced alpha-amylase secretion. Furthermore, anionic surfactants exhibited stabilizing effect on the enzyme during preservation at 4 degrees C. CONCLUSIONS: PEG 8000 and the ionic detergents (SDS, cholic acid and Tween-80) were more effective in the solubilization of cell membrane components, and enhancing enzyme yields than the cationic detergents such as CTAB (N,Cetyl-N,N,N-trimethyl ammonium bromide). Further, these surfactants were found to stabilize the enzyme at 4 degrees C. SIGNIFICANCE AND IMPACT OF THE STUDY: The secretion of Ca2+ independent hyperthermostable alpha-amylase was enhanced in the presence of certain anionic and non-ionic detergents in the medium. Furthermore, the surfactants stabilized the enzyme during preservation at 4 degrees C. The use of this enzyme in starch hydrolysis eliminates the addition of Ca2+ in starch liquefaction and its subsequent removal by ion exchange from sugar syrups. PMID- 12641710 TI - Development of a miniaturized four-culture method for the rapid enumeration of four bacterial groups in ground beef. AB - AIMS: A simplified and rapid method was developed to identify and enumerate total mesophilic microbial load, gram-negative bacteria, coliforms, and Escherichia coli in ground beef, using a single module. METHODS AND RESULTS: A 96-well microtiter plate was divided into four sections of two lanes (24 wells), with each section dedicated to each of the four critical bacterial groups. Double strength of four different selective broths (100 microl) was added to each well and an aliquot of sample (100 microl) from the first well was transferred to the next to achieve a twofold serial dilution in each series (24 wells). Initial bacterial load was calculated using the final consecutive positive reaction exhibiting color change or fluorescence. To validate the method, ground beef (n = 32), inoculated with a three-strain mixture of E. coli, together with 30 samples of commercial ground beef, were assayed by the four-culture method and conventional plating. For all media tested, the coefficients of determinations (r2) between the results derived from the four-culture method and those of conventional plating method ranged from 0.83 to 0.92. CONCLUSIONS: The high degree of association between the two methods indicates that the four-culture method could be easily applied to the enumeration of four critical bacterial groups in ground beef. SIGNIFICANCE AND IMPACT OF THE STUDY: This method will facilitate sanitation monitoring at meat processing plants by shortening time required for results, reducing consumable material costs, simplifying procedures, reducing the space required, and cutting the cost of start up equipment. PMID- 12641711 TI - Production and partial characterization of a beta-amylase by Xanthophyllomyces dendrorhous. AB - AIMS: The characterization of a beta-amylase produced by Xanthophyllomyces dendrorhous. METHODS AND RESULTS: Growth in different culture media showed that X. dendrorhous produces an amylase whose synthesis is repressed by the carbon source and induced by starch and maltose. Enzymatic assays using substrates with different degrees of polymerization together with viscosity experiments revealed that the enzyme was beta-amylase. According to the biochemical characterization, the enzyme has a molecular weight of 240 kDa and a Km of 1.35 mg ml-1. The optimum pH and temperature were 5.5 and 50 degrees C, respectively. Using different inhibitors of the enzymatic activity it was shown that cysteine, tryptophan and serine are essential amino acids for catalysis. CONCLUSIONS: Xanthophyllomyces dendrorhous CECT1690 synthesizes and secretes beta-amylase that could be a by-product, in addition to carotenoid pigments, in the fermentation downstream. SIGNIFICANCE AND IMPACT OF THE STUDY: The beta-amylase produced by X. dendrorhous may have certain industrial applications. PMID- 12641712 TI - Prevalence of bacterial faecal pathogens in separated and unseparated stored pig slurry. AB - AIMS: To examine the prevalence and diversity of bacterial faecal pathogens in unseparated slurry, separated solids and liquid fractions from a commercial pig farm. METHODS: A total of 43 stored slurry specimens originating from a fattening house over the period February-April 2002 were analysed, consisting of unseparated (n = 14) slurry, separated solids (n = 16) and separated liquid (n = 13). Specimens were examined for the presence of five bacterial pathogens including Salmonella spp., Shigella spp., Campylobacter spp., Escherichia coli O157 and Yersinia enterocolitica. Selective enrichment and plating methods were employed for detection of Salmonella spp. and Campylobacter spp. and conventional selective plating techniques for the remaining genera. Antibiogram profiles to 12 antibiotic agents were obtained for all Salmonella isolates obtained. RESULTS: Salmonella spp. were identified in all components of the slurry specimens, whereas Campylobacter spp. was only recovered from the unseparated and separated liquid fractions. In both cases, the separated liquid fraction had the highest prevalence of pathogens and the separated solid fraction had the lowest prevalence. None of the slurry specimens examined were positive for E. coli O157:H7, Shigella spp. or Y. enterocolitica. Twenty-nine isolates of Salmonella were recovered from the slurry specimens, comprising seven serovars, of which Salmonella manhattan was the most prevalent, accounting for over half [15 of 29 (51.7%)] of all Salmonella isolates. Salmonella anatum, Salm. derby, Salm. give, Salm. heidelberg, Salm. simi and Salm. stanley serovars were also recovered. All Salmonella isolates were sensitive to ampicillin, augmentin (amoxicillin/clavulanic acid), chloramphenicol, ciprofloxacin, gentamicin, kanamycin and trimethoprim, but has variable resistance to tetracycline (100%), sulphonamides (84.6%), furazolidone (38.5%), nalidixic acid (15.4%) and streptomycin (15.4%). The majority (57.7%) of isolates displayed antibiotic resistance to at least two antibiotic agents, followed by 34.6% of isolates being resistant to three agents and the remainder (7.7%) being resistant to four antibiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated a marked reduction in the prevalence of Campylobacter and Salmonella in the solids component of separated pig slurry. The adoption of control processes such as aeration of slurry prior to its spread onto agricultural land and newer approaches to pathogen reduction should be investigated, to reduce the transmission of pathogens from pig slurry to the environment. PMID- 12641713 TI - An in vitro model for investigating intestinal adhesion of potential dairy propionibacteria probiotic strains using cell line C2BBe1. AB - AIMS: The purposes of this study were to screen the adhesion properties of dairy propionibacteria strains and evaluate whether C2BBe1 could be used in the screening of potential probiotic strains. METHODS AND RESULTS: Thirteen dairy propionibacteria strains and two control strains, Lactobacillus acidophilus MJLA1 and Bifidobacterium lactis BDBB2, were tested for adhesion to C2BBe1. Electron microscopic observations demonstrated that the control strains, L. acidophilus MJLA1 and B. lactis BDBB2, had similar adhesive ability to C2BBe1 as had been previously shown to Caco-2. Only one of the 13 strains of dairy propionibacteria, strain P. jensenii 702, demonstrated adhesion to C2BBe1. CONCLUSIONS: C2BBe1 can provide an alternative to Caco-2 for assessing in vitro adhesion properties of probiotic strains. Adhesion properties of dairy propionibacteria were strain dependent. SIGNIFICANCE AND IMPACT OF THE STUDY: C2BBe1 is highly suitable for application in bacterial adhesion studies, and was used successfully to select a new potential probiotic. PMID- 12641714 TI - Evaluation of selective and non-selective enrichment PCR procedures for Salmonella detection. AB - AIMS: To compare PCR combined with enrichment media with the standard microbiological techniques (SMT) and to determine the most sensitive method for the detection of Salmonella and the identification of Salm. typhimurium (ST), Salm. enteritidis (SE), Salm. gallinarum (SG) and Salm. pullorum (SP). METHODS AND RESULTS: We analysed 87 samples from poultry using PCR and SMT, PCR being performed from non-selective (NS) and Rappaport-Vassiliadis (RV) media. PCR-NS was less sensitive than PCR-RV and SMT for the detection and identification of Salmonella. PCR-RV detected more positive samples of Salmonella sp. than SMT but both these methods showed similar sensitivity regarding the identification of Salmonella serovars. CONCLUSIONS: PCR-RV was more sensitive and decreased the time necessary to detect and identify Salmonella. SIGNIFICANCE AND IMPACT OF THE STUDY: PCR-RV is a powerful tool for the rapid and accurate detection and identification of Salmonella and can be implemented in diagnostic and food analysis laboratories. PMID- 12641715 TI - Molecular fingerprinting of Helicobacter pylori strains from duodenal ulcer patients. AB - AIMS: To characterize the molecular fingerprinting of Helicobacter pylori population isolated in duodenal ulcer patients treated with triple therapy. METHODS AND RESULTS: Gastric biopsy specimens from corpus and antrum, were cultured for H. pylori isolation. Helicobacter pylori eradication was evaluated after 4 and 16 weeks. DNAs of all isolates were characterized by random amplified polymorphic DNA typing and cagA gene was also detected. After the therapy, five patients harboured the microorganism at 4 weeks and two of them remained H. pylori positive at 16 weeks. The analysis of DNA fingerprinting of strains isolated from antrum and corpus of patients susceptible to treatment, showed similar patterns. Instead, when the therapy was not effective, strains isolated from sequential biopsies from initial and after 4 and 16 weeks, showed distinct fingerprintings and retained the cagA status, over time. CONCLUSIONS: The drugs used for therapy could exercise an effect in genotypical rearrangement among H. pylori cells. SIGNIFICANCE AND IMPACT OF THE STUDY: The variableness among H. pylori strains represents a way to challenge environmental stress. PMID- 12641716 TI - Proteolytic activity of a yeast cell wall lytic Arthrobacter species. AB - AIMS: To investigate properties of the proteolytic activity of a yeast cell wall lytic soil bacterium identified as an Arthrobacter species. METHODS AND RESULTS: The organism was grown at pH 7.5 and 30 degrees C in shake flasks on media with different complex subtrates. Highest proteolytic activity assayed with azocaseine was detected in media with wheat gluten. In addition, l-leucine, l-alanine exopeptidase activity and esterase activity were found. The proteolytic activity showed stability up to pH 12, with a maximum at pH 11. The temperature optimum was at 55 degrees C, but there was a loss in enzyme activity of 50% within 2 h. The proteolytic activity was inhibited by 3,4-dichloroisocumarin, whereas there was little or no effect with EDTA, pepstatin A or E64. CONCLUSIONS: The proteolytic activity is highly alkaline stable. The formation of the enzyme can be induced by media with high protein content. PMID- 12641717 TI - An improved selective isolation medium for the recovery of Listeria monocytogenes from smoked fish. AB - AIMS: The aim of this study was to improve the selective isolation of Listeria monocytogenes from smoked haddock fillets. METHODS AND RESULTS: Listeria selective agar (LSA)--Oxford formulation was supplemented with 25 microg x ml(-1) of colistin sulphate and 30 microg x ml(-1) of nalidixic acid. Inocula from four smoked haddock fillets produced colonies (approx. 2-13 bacteria x g(-1)), identified as L. monocytogenes, on LSA supplemented with antimicrobial compounds (MLSA). Moreover, there was only negligible evidence of bacteria which were not L. monocytogenes on MLSA. In contrast, LSA supported dense bacterial growth, which was not equated with L. monocytogenes. SIGNIFICANCE AND IMPACT OF THE STUDY: The modified medium permitted the recovery of L. monocytogenes from smoked haddock fillets and reduced the growth of contaminating bacteria. PMID- 12641718 TI - sanC--a novel gene involved in nikkomycin biosynthesis in Streptomyces ansochromogenes. AB - AIMS: To investigate the genes involved in the nikkomycin biosynthesis and their molecular mechanism. METHODS AND RESULTS: A 0.9 kbp SmaI fragment was cloned and sequenced which contains a complete open reading frame designated sanC (GenBank accession no. AF228522). In search of database, the deduced product of sanC was not homologous with any known proteins. The disruption and complementation of sanC showed that sanC is essential for nikkomycin biosynthesis in Streptomyces ansochromogenes. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: sanC is a novel and essential gene involved in nikkomycin biosynthesis in S. ansochromogenes. PMID- 12641719 TI - Improvement of Pseudomonas fluorescens CHA0 biocontrol activity against root-knot nematode by the addition of ammonium molybdate. AB - AIMS: To improve the efficacy of Pseudomonas fluorescens CHA0 and its genetically modified (GM) derivatives by adding ammonium molybdate to control Meloidogyne javanica, the root-knot nematode in mungbean. METHODS AND RESULTS: Culture filtrate of P. fluorescens CHA0 and its GM derivative (antibiotic overproducing strain CHA0/pME3424 and antibiotic-deficient CHA89) obtained from nutrient broth yeast extract medium amended with 1, 2 or 4 mm of ammonium molybdate (NH4-Mo) caused substantial mortality of M. javanica juveniles in vitro. Pseudomonas fluorescens CHA0 or CHA0/pME3424 applied in conjunction with NH4-Mo caused greater reduction of nematode penetration in mungbean roots compared with the bacterial application alone. Ammonium molybdate at 4 mg kg-1 of soil along with CHA0 also enhanced plant height while shoot weight remained unaffected. Either used alone or in conjunction with NH4-Mo, strain CHA89 did not reduce nematode invasion compared with the controls. Bacterial strains did not differ significantly in their colonization potential in the mungbean rhizosphere. Efficacy of the biocontrol bacteria to control root-knot nematode was accentuated when soil was treated with NH4-Mo and zinc (both at 1 mg kg-1 of soil). CONCLUSION: The addition of ammonium molybdate enhances the production of nematicidal compounds by P. fluorescensin vitro and improves bacterial efficacy against root-knot nematode under glasshouse conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Application of minerals such as ammonium molybdate is appealing because they are cheap and can easily be applied under field conditions to improve biocontrol potential of the bacterial inoculants. They also significantly reduce the amount of biocontrol inoculant biomass required to achieve root-knot disease control, with a consequent reduction in cost. PMID- 12641720 TI - Anomalies in species identification of enterococci from veterinary sources using a commercial biochemical identification system. AB - AIMS: A commercial biochemical panel ID kit was used to identify presumptive enterococci isolates of veterinary or agricultural origin obtained during different steps of culture. METHODS AND RESULTS: Fifty isolates identified as enterococci using a genus PCR assay were tested for genus and species identification using the BBL Crystal Identification Gram-Positive ID kit (Becton Dickinson, Sparks, MD, USA). Following sub-culture of the isolates three times, 59% agreement with the original panel ID was obtained. After four and six sub cultures, percentage agreement increased to 61 and 64%, respectively. Nineteen of the 50 cultures were identified as both Enterococcus faecalis and E. faecium. CONCLUSIONS: As a result of the variability between speciation of isolates following re-culture, additional methods for speciation are warranted. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests that the identification of the genus and species of non-human enterococcal isolates can vary greatly during successive passages when using this kit. PMID- 12641721 TI - Importance of Gram-positive naphthalene-degrading bacteria in oil-contaminated tropical marine sediments. AB - AIMS: The aim of this study was to isolate, characterize and evaluate the importance of naphthalene-degrading bacterial strains from oil-contaminated tropical marine sediments. METHODS AND RESULTS: Three Gram-positive naphthalene degrading bacteria were isolated from oil-contaminated tropical intertidal marine sediments by direct isolation or enrichment using naphthalene as the sole source of carbon and energy. Bacillus naphthovorans strain MN-003 can also grow on benzene, toluene, xylene and diesel fuel while Micrococcus sp. str. MN-006 can also grow on benzene. Staphylococcus sp. str. MN-005 can only degrade naphthalene and was not able to use the other aromatic hydrocarbons tested. Strain MN-003 possessed the highest maximal specific growth rate with naphthalene as sole carbon source. An enrichment culture fed with naphthalene as sole carbon source exhibited a significant increase in the relative abundances of the three isolates after 21 days of incubation. The three isolates constituted greater than 69% of the culturable naphthalene-degrading microbial community. Strain MN-003 outcompeted and dominated the other two isolates in competition studies involving batch cultures inoculated with equal cell densities of the three isolates and incubated with between 1 and 10 mg l-1 of naphthalene. CONCLUSIONS: Three Gram positive naphthalene-degrading bacteria were successfully isolated from oil contaminated tropical marine sediments. Gram-positive bacteria might play an important role in naphthalene degradation in the highly variable environment of oil-contaminated tropical intertidal marine sediments. Among the three isolates, strain MN-003 has the highest maximal specific growth rate when grown on naphthalene, and outgrew the other two isolates in competition experiments. SIGNIFICANCE AND IMPACT OF THE STUDY: This research will aid in the development of bioremediation schemes for oil-contaminated marine environments. Strain MN-003 could potentially be exploited in such schemes. PMID- 12641722 TI - The cortical serotonin2A receptor and the pathology of schizophrenia: a likely accomplice. AB - A large body of evidence shows that there is a change in the density of cortical serotonin2A receptors (5HT2AR) in post-mortem CNS from subjects with schizophrenia. Furthermore, some antipsychotic drugs have also been shown to cause a decrease in the density of 5HT2AR in the rat CNS. Thus, it appeared possible that changes in this receptor in human post-mortem CNS simply reflected an antipsychotic drug effect. However, a great deal of research on the 5HT2AR and schizophrenia now suggests that the changes in this receptor are complex and may be involved in both the pathology of the disorder and the effects of some antipsychotic drugs. Moreover, recent advances in basic research on the role of the 5HT2AR in the CNS add further support to the hypothesis that the receptor could be involved in the pathology of the illness. In particular, an argument will be developed that the changes in the 5HT2AR in schizophrenia are reflective of a real or perceived change in serotonergic tone and that this forms an important part of the pathology of the illness. PMID- 12641723 TI - The roles of calcium/calmodulin-dependent and Ras/mitogen-activated protein kinases in the development of psychostimulant-induced behavioral sensitization. AB - Although the development of behavioral sensitization to psychostimulants such as cocaine and amphetamine is confined mainly to one nucleus in the brain, the ventral tegmental area (VTA), this process is nonetheless complex, involving a complicated interplay between neurotransmitters, neuropeptides and trophic factors. In the present review we present the hypothesis that calcium-stimulated second messengers, including the calcium/calmodulin-dependent protein kinases and the Ras/mitogen-activated protein kinases, represent the major biochemical pathways whereby converging extracellular signals are integrated and amplified, resulting in the biochemical and molecular changes in dopaminergic neurons in the VTA that represent the critical neuronal correlates of the development of behavioral sensitization to psychostimulants. Moreover, given the important role of calcium-stimulated second messengers in the expression of behavioral sensitization, these signal transduction systems may represent the biochemical substrate through which the transient neurochemical changes associated with the development of behavioral sensitization are translated into the persistent neurochemical, biochemical and molecular alterations in neuronal function that underlie the long-term expression of psychostimulant-induced behavioral sensitization. PMID- 12641724 TI - Changes in dialysate concentrations of glutamate and GABA in the brain: an index of volume transmission mediated actions? AB - Brain microdialysis has become a frequently used method to study the extracellular concentrations of neurotransmitters in specific areas of the brain. For years, and this is still the case today, dialysate concentrations and hence extracellular concentrations of neurotransmitters have been interpreted as a direct index of the neuronal release of these specific neurotransmitter systems. Although this seems to be the case for neurotransmitters such as dopamine, serotonin and acetylcholine, the extracellular concentrations of glutamate and GABA do not provide a reliable index of their synaptic exocytotic release. However, many microdialysis studies show changes in extracellular concentrations of glutamate and GABA under specific pharmacological and behavioural stimuli that could be interpreted as a consequence of the activation of specific neurochemical circuits. Despite this, we still do not know the origin and physiological significance of these changes of glutamate and GABA in the extracellular space. Here we propose that the changes in dialysate concentrations of these two neurotransmitters found under specific treatments could be an expression of the activity of the neurone-astrocyte unit in specific circuits of the brain. It is further proposed that dialysate changes of glutamate and GABA could be used as an index of volume transmission mediated actions of these two neurotransmitters in the brain. This hypothesis is based firstly on the assumption that the activity of neurones is functionally linked to the activity of astrocytes, which can release glutamate and GABA to the extracellular space; secondly, on the existence of extrasynaptic glutamate and GABA receptors with functional properties different from those of GABA receptors located at the synapse; and thirdly, on the experimental evidence reporting specific electrophysiological and neurochemical effects of glutamate and GABA when their levels are increased in the extracellular space. According to this concept, glutamate and GABA, once released into the extracellular compartment, could diffuse and have long-lasting effects modulating glutamatergic and/or GABAergic neurone-astrocytic networks and their interactions with other neurotransmitter neurone networks in the same areas of the brain. PMID- 12641725 TI - delta-Opioid receptors exhibit high efficiency when activating trimeric G proteins in membrane domains. AB - Low-density membrane fragments (domains) were separated from the bulk of plasma membranes of human embryonic kidney (HEK)293 cells expressing a delta-opioid (DOP) receptor-Gi1alpha fusion protein by drastic homogenization and flotation on equilibrium sucrose density gradients. The functional activity of trimeric G proteins and capacity of the DOP receptor to stimulate both the fusion protein linked Gi1alpha and endogenous pertussis-toxin sensitive G proteins was measured as d-Ala2, d-Leu5-enkephalin stimulated high-affinity GTPase or guanosine-5' [gamma-35S]triphosphate ([35S]GTPgammaS) binding. The maximum d-Ala2-d-Leu5 enkephalin (DADLE)-stimulated GTPase was two times higher in low-density membrane fragments than in bulk of plasma membranes; 58 and 27 pmol/mg/min, respectively. The same difference was obtained for [35S]GTPgammaS binding. Contrarily, the low density domains contained no more than half the DOP receptor binding sites (Bmax = 6.6 pmol/mg versus 13.6 pmol/mg). Thus, when corrected for expression levels of the receptor, low-density domains exhibited four times higher agonist-stimulated GTPase and [35S]GTPgammaS binding than the bulk plasma membranes. The regulator of G protein signaling RGS1, enhanced further the G protein functional activity but did not remove the difference between domain-bound and plasma membrane pools of G protein. The potency of the agonist in functional studies and the affinity of specific [3H]DADLE binding to the receptor were, however, the same in both types of membranes - EC50 = 4.5 +/- 0.1 x 10(-8) and 3.2 +/- 1.4 x 10(-8) m for GTPase; Kd = 1.2 +/- 0.1 and 1.3 +/- 0.1 nm for [3H]DADLE radioligand binding assay. Similar results were obtained when sodium bicarbonate was used for alkaline isolation of membrane domains. By contrast, detergent-insensitive membrane domains isolated following treatment of cells with Triton X100 exhibited no DADLE-stimulated GTPase or GTPgammaS binding. Functional coupling between the DOP receptor and cognate G proteins was also blocked by high-energy ultrasound and repeated freezing-thawing. Our data indicate, for the first time, that membrane domains isolated using 'detergent-free' procedures exhibit higher efficiency of coupling between a G protein-coupled receptor and its corresponding G protein(s) than bulk plasma membranes. Detergent-extraction diminishes these interactions, even when the receptor and G proteins are physically tethered together. PMID- 12641726 TI - ASK1-p38 MAPK/JNK signaling cascade mediates anandamide-induced PC12 cell death. AB - Anandamide is a neuroimmunoregulatory molecule that triggers apoptosis in a number of cell types including PC12 cells. Here, we investigated the molecular mechanisms underlying anandamide-induced cell death in PC12 cells. Anandamide treatment resulted in the activation of p38 mitogen-activated protein kinase (MAPK), c-Jun N-terminal kinase (JNK), and p44/42 MAPK in apoptosing cells. A selective p38 MAPK inhibitor, SB203580, or dn-JNK, JNK1(A-F) or SAPKbeta(K-R), blocked anandamide-induced cell death, whereas a specific inhibitor of MEK-1/2, U0126, had no effect, indicating that activation of p38 MAPK and JNK is critical in anandamide-induced cell death. An important role for apoptosis signal regulating kinase 1 (ASK1) in this event was also demonstrated by the inhibition of p38 MAPK/JNK activation and death in cells overexpressing dn-ASK1, ASK1 (K709M). Conversely, the constitutively active ASK1, ASK1DeltaN, caused prolonged p38 MAPK/JNK activation and increased cell death. These indicate that ASK1 mediates anandamide-induced cell death via p38 MAPK and JNK activation. Here, we also found that activation of p38 MAPK/JNK is accompanied by cytochrome c release from the mitochondria and caspase activation (which can be inhibited by SB203580), suggesting that anandamide triggers a mitochondrial dependent apoptotic pathway. The caspase inhibitor, zVAD, and the mitochondrial pore opening inhibitor, cyclosporine A, blocked anandamide-induced cell death but not p38 MAPK/JNK activation, suggesting that activation of these kinases may occur upstream of mitochondrial associated events. PMID- 12641728 TI - Immunocytochemical localization of glycogen phosphorylase isozymes in rat nervous tissues by using isozyme-specific antibodies. AB - Isozyme-specific antibodies were raised against peptides from the low-homology regions of the sequences of rat glycogen phosphorylase BB and MM isozymes by immunization of rabbits and guinea pigs. Immunocytochemical double-labelling experiments on frozen sections of rat nervous tissues were performed to investigate the isozyme localization pattern. Astrocytes throughout the brain and spinal cord expressed both isozymes in perfect co-localization. Ependymal cells only expressed the BB isozyme. Most neurones were not immunoreactive. The rare neurones that contained glycogen phosphorylase only expressed the BB isozyme. Nearly all of these neurones formed part of the afferent somatosensory system. These findings stress the general importance of glycogen in neural energy metabolism and indicate a special role for the glycogen phosphorylase BB isozyme in neurones in the somatosensory system. PMID- 12641727 TI - Contribution of two conserved glycine residues to fibrillogenesis of the 106-126 prion protein fragment. Evidence that a soluble variant of the 106-126 peptide is neurotoxic. AB - The fibrillogenic peptide corresponding to the residues 106-126 of the prion protein sequence (PrP 106-126) is largely used to explore the neurotoxic mechanisms underlying the prion disease. However, whether the neuronal toxicity of PrP 106-126 is caused by a soluble or fibrillar form of this peptide is still unknown. The aim of this study was to correlate the structural state of this peptide with its neurotoxicity. Here we show that the two conserved Gly114 and Gly119 residues, in force of their intrinsic flexibility, prevent the peptide assuming a structured conformation, favouring its aggregation in amyloid fibrils. The substitution of both Gly114 and Gly119 with alanine residues (PrP 106-126 AA mutated peptide) reduces the flexibility of this prion fragment and results in a soluble, beta-structured peptide. Moreover, PrP 106-126 AA fragment was highly toxic when incubated with neuroblastoma cells, likely behaving as a neurotoxic protofibrillar intermediate of the wild-type PrP 106-126. These data further confirm that the fibrillar aggregation is not necessary for the induction of the toxic effects of PrP 106-126. PMID- 12641730 TI - JKK1, an upstream activator of JNK/SAPK, is activated in Alzheimer's disease. AB - JNK/SAPK has been implicated in the pathogenesis of Alzheimer's disease, but the upstream cascade leading to JNK/SAPK activation has not been elucidated in the disease. In this study, we focused on one of the physiological activators of JNK/SAPK, JNK kinase 1 (JKK1). Although there was no significant difference in the level and distribution of total JKK1 between Alzheimer's disease (AD) and age matched control cases, increased levels of activated phospho-JKK1 were specifically localized to neurofibrillary pathology including neurofibrillary tangles, senile plaque neurites, granulovaualar degenerations and neuropil threads in severe AD (Braak stage V-VI), considerably overlapping with its downstream effector, phospho-JNK/SAPK, suggesting both a functional and mechanistic link. Nuclear localization of phospho-JKK1 was also found in mild (Braak stage III-IV) but not in severe AD cases (Braak stage V-VI), suggesting a possible re-distribution correlating with the progress of the disease. By immunoblot analyses, phospho-JKK1 was significantly increased in AD over control cases. Together, these findings lend further credence to the notion that the JNK/SAPK pathway is dysregulated in AD and also indicate an active role for this pathway in disease pathogenesis. PMID- 12641729 TI - Effects of L-dopa and other amino acids against paraquat-induced nigrostriatal degeneration. AB - Exposure to the herbicide paraquat causes selective nigrostriatal degeneration and aggregation of alpha-synuclein in the mouse brain. The purpose of this study was to assess mechanisms of paraquat entry into the CNS and, in particular, the effects of substrates of the blood-brain barrier (BBB) neutral amino acid transporter (System L carrier) on paraquat accumulation and neurotoxicity. Using a paraquat antibody, robust immunoreactivity was observed in the midbrain of mice injected with the herbicide. This immunoreactivity was abolished by administration of l-valine or l-phenylalanine, two System L substrates, immediately before paraquat exposure. Pre-treatment with these amino acids completely protected against paraquat-induced loss of nigrostriatal dopaminergic cells and formation of thioflavine S-positive intracellular deposits. Interestingly, the anti-parkinsonian drug l-dopa, which is transported across the BBB through the same neutral amino acid carrier, was also neuroprotective when administered 30 min prior to paraquat. In contrast, paraquat-induced toxicity was unaffected if animals (i) were pre-treated with d-valine, the biologically inactive d-isomer of l-valine, or with l-lysine, a substrate of the basic rather than the neutral amino acid carrier, or (ii) were injected with l-dopa 24 h after paraquat exposure. Data are consistent with a critical role of uptake across the BBB in paraquat neurotoxicity, and suggest that dietary elements (e.g. amino acids) or therapeutic agents (e.g. l-dopa) may modify the effects of toxicants targeting the nigrostriatal system. PMID- 12641731 TI - Behavioural and gene transcription alterations induced by spontaneous cannabinoid withdrawal in mice. AB - This study examined behavioural signs that occur during tolerance development to cannabinoid treatment and hormonal and gene expression alterations induced by spontaneous cannabinoid withdrawal in mice. Tolerance to CP-55,940 treatment developed for hypothermia, ambulatory and exploratory locomotor activity. Cessation of cannabinoid treatment resulted in a behavioural withdrawal syndrome characterized by a pronounced increase in ambulatory activity and rearings. Corticosterone plasma concentrations dramatically increased 24 and 72 h after cessation of cannabinoid treatment. Similarly, an increase (40%) in cannabinoid [35S]GTPgammaS binding autoradiography was detected on days 1 and 3 of abstinence. Spontaneous cannabinoid withdrawal produced time-related significant alterations in gene transcription: (i) decreased (20%) tyrosine hydroxylase (TH) mRNA levels in the ventral tegmental area and increased (50%) in substantia nigra; (ii) increased proenkephalin (PENK) gene expression more than 100% in caudate-putamen, nucleus accumbens, olfactory tubercle and piriform cortex; (iii) increased (20-40%) pro-opiomelanocortin (POMC) gene expression in the arcuate nucleus of the hypothalamus. These results suggest that spontaneous cannabinoid withdrawal occur after cessation of CP-55,940 treatment. This 'syndrome' includes behavioural, hormonal and gene transcription alterations that seems to be part of the regulation of neuronal plasticity induced by spontaneous cannabinoid withdrawal. PMID- 12641732 TI - Amphetamine-induced c-fos mRNA expression in the caudate-putamen and subthalamic nucleus: interactions between dose, environment, and neuronal phenotype. AB - When administered in a novel environment relatively low doses of amphetamine induce c-fos mRNA in the subthalamic nucleus (STN) and in preproenkephalin mRNA containing (ENK+) neurons in the caudate-putamen (CPu). When administered at home, however, low doses of amphetamine do not produce these effects. Environmental novelty also facilitates the behavioral effects of acute and repeated amphetamine, but this is dose-dependent. The purpose of the present experiment therefore was to determine if the effect of context on amphetamine induced c-fos expression is also dose-dependent. It was found that: (i) No dose of amphetamine tested (1-10 mg/kg) induced c-fos in many ENK+ cells when given at home. (ii) When given in a novel environment low to moderate doses of amphetamine (1-5 mg/kg) induced c-fos in substantial numbers of ENK+ cells, but the highest dose examined (10 mg/kg) did not. (iii) Environmental novelty enhanced the ability of low to moderate doses of amphetamine to induce c-fos in the STN, but the highest dose of amphetamine induced robust c-fos mRNA expression in the STN regardless of context. The results do not support the idea that engaging ENK+ cells, at least as indicated by c-fos mRNA expression, is critical to produce robust behavioral sensitization, but do suggest a possible role for the STN. Furthermore, the results highlight the importance of drug-environment interactions on the neurobiological effects of drugs, and have implications for thinking about the circuits by which context modulates the acute and long-lasting consequences of amphetamine treatment. PMID- 12641733 TI - Proteasome inhibition by paired helical filament-tau in brains of patients with Alzheimer's disease. AB - Alzheimer's disease (AD) is characterized neuropathologically by intracellular neurofibrillary tangles (NFTs) formed of tau-based paired helical filaments (PHFs) and extracellular beta-amyloid plaques. The degree of Alzheimer dementia correlates with the severity of PHFs and NFTs. As an intraneuronal accumulation of oxidatively damaged proteins has been found in the brains of patients with AD, a dysfunction of the proteasomal system, which degrades damaged proteins, has been assumed to cause protein aggregation and therefore neurodegeneration in AD. In this study, we revealed that such proteasome dysfunction in AD brain results from the inhibitory binding of PHF-tau to proteasomes. We analysed the proteasome activity in brains from patients with AD and age-matched controls, and observed a significant decrease to 56% of the control level in the straight gyrus of patients with AD. This loss of activity was not associated with a decrease in the proteasome protein. PHF-tau co-precipitated during proteasome immunoprecipitation and proteasome subunits could be co-isolated during isolation of PHFs from AD brain. Furthermore, the proteasome activity in human brains strongly correlated with the amount of co-precipitated PHF-tau during immunoprecipitation of proteasome. Incubation of isolated proteasomes with PHF-tau isolated from AD brain, and with PHFs after in vitro assembly from human recombinant tau protein, resulted in a distinct inhibition of proteasome activity by PHF-tau. As this inhibition of proteasome activity was sufficient to induce neuronal degeneration and death, we suggest that PHF-tau is able directly to induce neuronal damage in the AD brain. PMID- 12641734 TI - CIP98, a novel PDZ domain protein, is expressed in the central nervous system and interacts with calmodulin-dependent serine kinase. AB - Receptors and various molecules in neurons are localized at precise locations to perform their respective functions, especially in synaptic sites. Among synaptic molecules, PDZ domain proteins play major roles in scaffolding and anchoring membrane proteins for efficient synaptic transmission. In the present study, we isolated CIP98, a novel protein (98 kDa) consisting of three PDZ domains and a proline-rich region, which is widely expressed in the central nervous system. In situ hybridization and immunohistochemical staining patterns demonstrate that CIP98 is expressed strongly in certain types of neurons, i.e. pyramidal cells in layers III-V of the cerebral cortex, projecting neurons in the thalamus and interneurons in the cerebellum. The results of immunocytochemical staining and electron microscopy revealed that CIP98 is localized both in dendrites and axons. Interestingly, CIP98 interacts with CASK (calmodulin-dependent serine kinase), a member of the membrane-associated guanylate kinase (MAGUK) family that plays important roles in the molecular organization of proteins at synapses. CIP98 was shown to co-localize with CASK along the dendritic processes of neurons. In view of its direct association with CASK, CIP98 may be involved in the formation of CASK scaffolding proteins complex to facilitate synaptic transmission in the CNS. PMID- 12641735 TI - Ethanol specifically decreases peroxisome proliferator activated receptor beta in B12 oligodendrocyte-like cells. AB - Peroxisome proliferator activated receptors (PPARs) are nuclear receptors that control important genes involved in lipid metabolism. Their role in nerve cells is uncertain, although anomalous myelination of the corpus callosum has been described in the PPARbeta-null mouse, and abnormalities of this tissue have been documented in fetal alcohol syndrome in humans. We report here that ethanol treatment of B12 oligodendrocyte-like cells induces a concentration- and time dependent decrease in the mRNA and protein levels of PPARbeta, with no effect on PPARalpha or PPARgamma. The effect on PPARbeta is seen as an increase in mRNA degradation, as assessed by run-off assays, due to a significant decrease in PPARbeta mRNA half-life, with no observed changes in intracellular localization. Our results suggest a possible link between PPARbeta function and ethanol-induced abnormal myelination in oligodendrocytes. PMID- 12641737 TI - Neuronal activity-dependent nucleocytoplasmic shuttling of HDAC4 and HDAC5. AB - The class II histone deacetylases, HDAC4 and HDAC5, directly bind to and repress myogenic transcription factors of the myocyte enhancer factor-2 (MEF-2) family thereby inhibiting skeletal myogenesis. During muscle differentiation, repression of gene transcription by MEF-2/HDAC complexes is relieved due to calcium/calmodulin-dependent (CaM) kinase-induced translocation of HDAC4 and HDAC5 to the cytoplasm. MEF-2 proteins and HDACs are also highly expressed in the nervous system and have been implicated in neuronal survival and differentiation. Here we investigated the possibility that the subcellular localization of HDACs, and thus their ability to repress target genes, is controlled by synaptic activity in neurones. We found that, in cultured hippocampal neurones, the localization of HDAC4 and HDAC5 is dynamic and signal-regulated. Spontaneous electrical activity was sufficient for nuclear export of HDAC4 but not of HDAC5. HDAC5 translocation to the cytoplasm was induced following stimulation of calcium flux through synaptic NMDA receptors or L-type calcium channels; glutamate bath application (stimulating synaptic and extrasynaptic NMDA receptors) antagonized nuclear export. Activity-induced nucleocytoplasmic shuttling of both HDACs was partially blocked by the CaM kinase inhibitor KN-62 with HDAC5 nuclear export being more sensitive to CaM kinase inhibition than that of HDAC4. Thus, the subcellular localization of HDACs in neurones is specified by neuronal activity; differences in the activation thresholds for HDAC4 and HDAC5 nuclear export provides a mechanism for input-specific gene expression. PMID- 12641736 TI - Iron porphyrin treatment extends survival in a transgenic animal model of amyotrophic lateral sclerosis. AB - Oxidative damage, produced by mutant Cu/Zn superoxide dismutase (SOD1), may play a role in the pathogenesis of amyotrophic lateral sclerosis (ALS), a devastating motor neuron degenerative disease. A novel approach to antioxidant therapy is the use of metalloporphyrins that catalytically scavenge a wide range of reactive oxygen and reactive nitrogen species. In this study, we examined the therapeutic potential of iron porphyrin (FeTCPP) in the G93A mutant SOD1 transgenic mouse model of ALS. We found that intraperitoneal injection of FeTCPP significantly improved motor function and extended survival in G93A mice. Similar results were seen with a second group of mice wherein treatment with FeTCPP was initiated at the onset of hindlimb weakness-roughly equivalent to the time at which treatment would begin in human patients. FeTCPP-treated mice also showed a significant reduction in levels of malondialdehyde (a marker of lipid peroxidation), in total content of protein carbonyls (a marker of protein oxidation), and increased neuronal survival in the spinal cord. These results therefore provide further evidence of oxidative damage in a mouse model of ALS, and suggest that FeTCPP could be beneficial for the treatment of ALS patients. PMID- 12641738 TI - Aggregate formation and the impairment of long-term synaptic facilitation by ectopic expression of mutant huntingtin in Aplysia neurons. AB - Huntington's disease (HD) is caused by an expansion of a polyglutamine (polyQ) tract within huntingtin (htt) protein. To examine the cytotoxic effects of polyQ expanded htt, we overexpressed an enhanced green fluorescent protein (EGFP) tagged N-terminal fragment of htt with 150 glutamine residues (Nhtt150Q-EGFP) in Aplysia neurons. A combined confocal and electron microscopic study showed that Aplysia neurons expressing Nhtt150Q-EGFP displayed numerous abnormal aggregates (diameter 0.5-5 microm) of filamentous structures, which were formed rapidly (approximately 2 h) but which were sustained for at least 18 days in the cytoplasm. Furthermore, the overexpression of Nhtt150Q-EGFP in sensory cells impaired 5-hydroxytryptamine (5-HT)-induced long-term synaptic facilitation in sensori-motor synapses without affecting basal synaptic strength or short-term facilitation. This study demonstrates the stability of polyQ-based aggregates and their specific effects on long-term synaptic plasticity. PMID- 12641739 TI - Dephosphorylation of pCREB by protein serine/threonine phosphatases is involved in inactivation of Aanat gene transcription in rat pineal gland. AB - The rat pineal gland is a suitable model to investigate neurotransmitter controlled gene expression, because it is well established that the stimulation of melatonin biosynthesis by norepinephrine (NE) depends on the activation of the gene that encodes arylalkylamine N-acetyltransferase (AANAT), the melatonin rhythm enzyme. The mechanisms responsible for downregulation of Aanat transcription are less clear. In this in vitro study we investigated the role of pCREB dephosphorylation for termination of Aanat gene transcription. Immunosignals for pCREB, strongly induced after NE stimulation, rapidly decreased after withdrawal of NE. The immunoreactivity of the inhibitory transcription factor ICER increased twofold after NE treatment for 6 h, but did not change within 30 min after removal of the stimulus. Application of protein serine/threonine phosphatase (PSP) inhibitors prevented pCREB dephosphorylation and blocked the decreases in Aanat mRNA levels, AANAT protein amount and melatonin biosynthesis all of which occurred rapidly after NE withdrawal. PSPs in the rat pineal gland were characterized by immunocytochemistry and immunoblotting. NE-stimulation for 8 h induced accumulation of PSP1-catalytic subunit (CSU) in pinealocyte nuclei, but did not affect the distribution of PSP2A CSU. The results identify dephosphorylation of pCREB by PSPs as an essential mechanism for downregulation of Aanat transcription in the rat pineal gland. PMID- 12641740 TI - Interaction between flavonoids and the blood-brain barrier: in vitro studies. AB - There is considerable current interest in the neuroprotective effects of flavonoids. This study focuses on the potential for dietary flavonoids, and their known physiologically relevant metabolites, to enter the brain endothelium and cross the blood-brain barrier (BBB) using well-established in vitro models (brain endothelial cell lines and ECV304 monolayers co-cultured with C6 glioma cells). We report that the citrus flavonoids, hesperetin, naringenin and their relevant in vivo metabolites, as well as the dietary anthocyanins and in vivo forms, cyanidin-3-rutinoside and pelargonidin-3-glucoside, are taken up by two brain endothelial cell lines from mouse (b.END5) and rat (RBE4). In both cell types, uptake of hesperetin and naringenin was greatest, increasing significantly with time and as a function of concentration. In support of these observations we report for the first time high apparent permeability (Papp) of the citrus flavonoids, hesperetin and naringenin, across the in vitro BBB model (apical to basolateral) relative to their more polar glucuronidated conjugates, as well as those of epicatechin and its in vivo metabolites, the dietary anthocyanins and to specific phenolic acids derived from colonic biotransformation of flavonoids. The results demonstrate that flavonoids and some metabolites are able to traverse the BBB, and that the potential for permeation is consistent with compound lipophilicity. PMID- 12641741 TI - Fluorescent two-dimensional difference gel electrophoresis and mass spectrometry identify age-related protein expression differences for the primary visual cortex of kitten and adult cat. AB - The recent introduction of fluorescent two-dimensional difference gel electrophoresis, combined with mass spectrometry, has greatly simplified the analysis and identification of differentially expressed proteins by eliminating intergel variability. In this report, we describe the successful application of this functional proteomics approach to compare protein expression levels in visual cortical area 17 of adult cats and 30-day-old kittens, in order to identify proteins expressed in an age-related fashion. We identified 16 proteins that were more abundantly expressed in kitten striate cortex and 12 proteins with a pronounced expression in adult cat area 17. Among those isolated from kitten area 17 were proteins related to axon growth and growth cone guidance and to the formation of cytoskeletal filaments. Glial fibrillary acidic protein, as identified in adult cat area 17, has been implicated previously in the termination of the critical period for cortical plasticity in kittens. In situ hybridization experiments for two of the identified proteins, glial fibrillary acidic protein and collapsin response mediator protein 5, confirmed and extended their differential expression to the mRNA level. Our findings show that two dimensional difference gel electrophoresis combined with mass spectrometry is a powerful approach that permits the identification of small protein expression differences correlated to different physiological conditions. PMID- 12641742 TI - Glutamate regulation of non-quantal release of acetylcholine in the rat neuromuscular junction. AB - Glutamate, previously demonstrated to participate in regulation of the resting membrane potential in skeletal muscles, also regulates non-quantal acetylcholine (ACh) secretion from rat motor nerve endings. Non-quantal ACh secretion was estimated by the amplitude of endplate hyperpolarization (H-effect) following blockade of skeletal muscle post-synaptic nicotinic receptors by (+)-tubocurarine and cholinesterase by armin (diethoxy-p-nitrophenyl phosphate). Glutamate was shown to inhibit non-quantal release but not spontaneous and evoked quantal secretion of ACh. Glutamate-induced decrease of the H-effect was enhanced by glycine. Glycine alone also lowered the H-effect, probably due to potentiation of the effect of endogenous glutamate present in the synaptic cleft. Inhibition of N methyl-d-aspartate (NMDA) receptors with (+)-5-methyl-10,11-dihydro-5H dibenzocyclohepten-5,10-imine (MK801), dl-2-amino-5-phosphopentanoic acid (AP5) and 7-chlorokynurenic acid or the elimination of Ca2+ from the bathing solution prevented the glutamate-induced decrease of the H-effect with or without glycine. Inhibition of muscle nitric oxide synthase by NG-nitro-l-arginine methyl ester (l NAME), soluble guanylyl cyclase by 1H[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) and binding and inactivation of extracellular nitric oxide (NO) by haemoglobin removed the action of glutamate and glycine on the H-effect. The results suggest that glutamate, acting on post-synaptic NMDA receptors to induce sarcoplasmic synthesis and release of NO, selectively inhibits non-quantal secretion of ACh from motor nerve terminals. Non-quantal ACh is known to modulate the resting membrane potential of muscle membrane via control of activity of chloride transport and a decrease in secretion of non-quantal transmitter following muscle denervation triggers the early post-denervation depolarization of muscle fibres. PMID- 12641744 TI - The adhesion protein TAG-1 has a ganglioside environment in the sphingolipid enriched membrane domains of neuronal cells in culture. AB - We studied the interactions between gangliosides and proteins at the exoplasmic surface of the sphingolipid-enriched membrane domains by ganglioside photolabeling combined with cell surface biotin labeling. After cell photolabeling with radioactive photoactivable derivatives of GM3, GM1 and GD1b gangliosides, followed by cell surface biotin labeling, sphingolipid-enriched domains were prepared and immunoprecipitated with streptavidin-coupled beads, under experimental conditions preserving the integrity of the lipid domain. About 50% of the total radioactivity linked to proteins was associated with acylated tubulin, about 10% with a 135-kDa protein present as a series of species with pI ranging from 6.5 to 8.0, about 5% with a protein of about 70 kDa and with pI near to 6.5. By immunoprecipitation with streptavidin-coupled beads under conditions disrupting the integrity of the lipid domain, the 135 kDa protein was recovered in the immunoprecipitate, that did not contain tubulin. Thus, the 135 kDa protein has an exoplasmic domain, and it was then identified as the GPI-anchored neural cell adhesion molecule TAG-1. Remarkably, TAG-1 was cross-linked in a similar extent by the photoactivated ganglioside GM3, GM1 and GD1b. The three gangliosides bear different oligosaccharide chains, suggesting that the ganglioside/TAG-1 interaction is not specifically associated with the ganglioside oligosaccharide structure. PMID- 12641743 TI - Tetrahydrobiopterin precursor sepiapterin provides protection against neurotoxicity of 1-methyl-4-phenylpyridinium in nigral slice cultures. AB - Complex-I inhibition and oxidative processes have been implicated in the loss of nigral dopamine neurones in Parkinson's disease and the toxicity of MPTP and its metabolite MPP+. Tetrahydrobiopterin, an essential cofactor for tyrosine hydroxylase, may act as an antioxidant in dopaminergic neurones and protects against the toxic consequences of glutathione depletion. Here we studied the effects of manipulating tetrahydrobiopterin levels on MPP+ toxicity in organotypic, rat ventral mesencephalic slice cultures. In cultures exposed to 30 micro m MPP+ for 2 days, followed by 8 days 'recovery' in control medium, we measured dopamine and its metabolites in the tissue and culture medium by HPLC, lactate dehydrogenase release to the culture medium, cellular uptake of propidium iodide and counted the tyrosine hydroxylase-immunoreactive neurones. Inhibition of tetrahydrobiopterin synthesis by 2,4-diamino-6-hydroxypyrimidine had no significant synergistic effect on MPP+ toxicity. In contrast, the tetrahydrobiopterin precursor l-sepiapterin attenuated the MPP+-induced dopamine depletion and loss of tyrosine hydroxylase-positive cells in a dose-dependent manner with 40 micro m l-sepiapterin providing maximal protection. Accordingly, increasing intracellular tetrahydrobiopterin levels may protect against oxidative stress by complex-I inhibition. PMID- 12641746 TI - Peripherin-mediated death of motor neurons rescued by overexpression of neurofilament NF-H proteins. AB - In previous studies, we showed that overexpression of peripherin, a neuronal intermediate filament (IF) protein, in mice deficient for neurofilament light (NF L) subunits induced a progressive adult-onset degeneration of spinal motor neurons characterized by the presence of IF inclusion bodies reminiscent of axonal spheroids found in amyotrophic lateral sclerosis (ALS). In contrast, the overexpression of human neurofilament heavy (NF-H) proteins provoked the formation of massive perikaryal IF protein accumulations with no loss of motor neurons. To further investigate the toxic properties of IF protein inclusions, we generated NF-L null mice that co-express both peripherin and NF-H transgenes. The axonal count in L5 ventral roots from 6 and 8-month-old transgenic mice showed that NF-H overexpression rescued the peripherin-mediated degeneration of motor neurons. Our analysis suggests that the protective effect of extra NF-H proteins is related to the sequestration of peripherin into the perikaryon of motor neurons, thereby abolishing the development of axonal IF inclusions that might block transport. These findings illustrate the importance of IF protein stoichiometry in formation, localization and toxicity of neuronal inclusion bodies. PMID- 12641747 TI - Social isolation-induced increase in the sensitivity of rats to the steroidogenic effect of ethanol. AB - Social isolation of rats for 30 days immediately after weaning results in marked decreases in the cerebrocortical and plasma concentrations of pregnenolone, progesterone, 3alpha-hydroxy-5alpha-pregnan-20-one (3alpha,5alpha-TH PROG), and 3alpha,5alpha-tetrahydrodeoxycorticosterone (3alpha,5alpha-TH DOC), as well as a moderate increase in the plasma concentration of corticosterone. This mildly stressful condition has now been shown to increase the sensitivity of rats to the effect of acute ethanol administration on the cerebrocortical and plasma concentrations of neuroactive steroids. The percentage increases in the brain and plasma concentrations of pregnenolone, progesterone, 3alpha,5alpha-TH PROG, and 3alpha,5alpha-TH DOC, apparent 20 min after a single intraperitoneal injection of ethanol (1 g/kg), were thus markedly greater in isolated rats than in group housed animals. A subcutaneous injection of isoniazid (300 mg/kg) also induced greater percentage increases in the concentrations of these steroids in isolated rats than in group-housed animals. These results suggest that mild chronic stress, such as that induced by social isolation, enhances the steroidogenic effect of ethanol, a drug abused by humans under stress or affected by neuropsychiatric disorders. Social isolation also induced hyper-responsiveness of the hypothalamic-pituitary-adrenal (HPA) axis, as was apparent after reduction of GABA-mediated inhibitory tone by isoniazid administration. PMID- 12641745 TI - Early events of target deprivation/axotomy-induced neuronal apoptosis in vivo: oxidative stress, DNA damage, p53 phosphorylation and subcellular redistribution of death proteins. AB - The mechanisms of injury- and disease-associated apoptosis of neurons within the CNS are not understood. We used a model of cortical injury in rat and mouse to induce retrograde neuronal apoptosis in thalamus. In this animal model, unilateral ablation of the occipital cortex induces apoptosis of corticopetal projection neurons in the dorsal lateral geniculate nucleus (LGN), by 7 days post lesion, that is p53 modulated and Bax dependent. We tested the hypothesis that this degenerative process is initiated by oxidative stress and early formation of DNA damage and is accompanied by changes in the levels of pro-apoptotic mediators of cell death. Immunoblotting revealed that the protein profiles of Bax, Bak and Bad were different during the progression of neuronal apoptosis in the LGN. Bax underwent a subcellular redistribution by 1 day post-lesion, while Bak increased later. Bad showed an early sustained increase. Cleaved caspase-3 was elevated maximally at 5 and 6 days. Active caspase-3 underwent a subcellular translocation to the nucleus. A dramatic phosphorylation of p53 was detected at 4 days post lesion. DNA damage was assessed immunocytochemically as hydroxyl radical adducts (8-hydroxy-2-deoxyguanosine) and single-stranded DNA. Both forms of DNA damage accumulated early in target-deprived LGN neurons. Transgenic overexpression of superoxide dismutase-1 provided significant protection against the apoptosis but antioxidant pharmacotreatments with trolox and ascorbate were ineffective. We conclude that overlapping and sequential signaling pathways are involved in the apoptosis of adult brain neurons and that DNA damage generated by superoxide derivatives is an upstream mechanism for p53-regulated, Bax-dependent apoptosis of target-deprived neurons. PMID- 12641748 TI - Xenobiotic response element binding enriched in both nuclear and microsomal fractions of rat cerebellum. AB - Xenobiotic response element (XRE) is a core nucleotide sequence at the upstream of inducible target genes for the transcription factor aryl hydrocarbon receptor (AhR) that is responsible for signal transduction of exogenous environmental pollutants in eukaryotic cells. Immunoblotting analysis revealed the constitutive expression of AhR-related proteins in rat liver and brain, while specific binding of a radiolabelled probe containing XRE was detected in nuclear preparations of both liver and brain on gel retardation electrophoresis. Among discrete rat brain structures examined, cerebellum exhibited the highest XRE binding with less potent binding in hypothalamus, midbrain, medulla-oblongata, hippocampus, cerebral cortex and striatum. In contrast to liver and hippocampus, cerebellum also contained unusually higher XRE binding in microsomal fractions than that in either nuclear or mitochondrial fractions. Limited proteolysis by V8 protease did not markedly affect XRE binding in cerebellar nuclear extracts, with concomitant diminution of that in hepatic and hippocampal nuclear extracts. In primary cultured cerebellar neurons, indigo was effective in significantly increasing XRE binding only when determined immediately after sustained exposure for 120 min in the presence of high potassium chloride. These results suggest the abundance of as-yet unidentified proteins with high affinity for XRE and responsiveness to indigo in both nuclear and microsomal fractions of rat cerebellum. PMID- 12641749 TI - Astrocyte metabolism is disturbed in the early development of experimental hydrocephalus. AB - The proper diagnosis of the arrested or the progressive form of hydrocephalus has a critical impact on treatment, but remains difficult. The assessment of early changes in cerebral metabolism might help in the development of adequate non invasive diagnostic tools. This study examined the alterations in label incorporation in neurotransmitter amino acids and other compounds in kaolin induced progressive hydrocephalus in rats by means of magnetic resonance spectroscopy (MRS) combined with the administration of [1-13C]glucose and [1,2 13C]acetate. Some 2, 4 and 6 weeks after kaolin injection into the cisterna magna, cerebrum, brainstem and cerebellum were dissected. Interestingly, labelling of most amino acids derived from [1-13C]glucose showed no alterations, whereas labelling from [1,2-13C]acetate was affected. Two weeks after induction of hydrocephalus the taurine concentration was decreased, whereas the concentration of [1,2-13C]lactate was increased in the cerebrum and that of [1,2 13C]GABA in the brainstem. Furthermore, labelling from [1,2-13C]acetate was significantly decreased in [4,5-13C]glutamate, [1,2-13C]glutamate and [1,2 13C]GABA in cerebrum from 4 weeks after hydrocephalus induction. The concentration of N-acetylaspartate, a neuronal marker, was unchanged. However, labelling of the acetyl group from [1-13C]glucose was decreased in cerebellum and brainstem at 6 weeks after the induction of hydrocephalus. As glucose is metabolized predominately by neurones, whereas acetate is exclusively taken up by astrocytes, these results indicate that mostly astrocytic, and only later neuronal, metabolism is disturbed in the kaolin model of hydrocephalus. If verified in patients using in vivo MRS, impaired astrocyte metabolism might serve as an early indication for operative treatment. PMID- 12641750 TI - Regulation of neuritogenesis and synaptic transmission by msec7-1, a guanine nucleotide exchange factor, in cultured Aplysia neurons. AB - msec7-1, a mammalian homologue of yeast sec7p, is known as a GDP/GTP exchange factor (GEF) for the ADP ribosylation factor (ARF) family of small GTPases. Here, we report that msec7-1 overexpression in cultured Aplysia neurons leads to an extensive neuritogenesis in a GEF activity-dependent manner through the modulation of the actin cytoskeleton. Similarly, the overexpression of ARNO, another mammalin GEF, produces extensive neuritogenesis in Aplysia neurons. In addition, msec7-1 overexpression increases the number of varicosities with an altered size and shape in a GEF activity-dependent manner. The overexpression of msec7-1 in pre-synaptic sensory neurons co-cultured with post-synaptic target motor neurons leads to an increase in the amplitude of the excitatory post synaptic potential through its GEF activity. Our results demonstrate that msec7-1 regulates neuritogenesis and synaptic transmission. PMID- 12641752 TI - Proceedings of the NFO-7, the International Conference on Near-field Optics and Related Techniques. University of Rochester, USA. 11-15 August 2002. PMID- 12641754 TI - Apertureless scanning near-field optical microscopy for ion exchange channel waveguide characterization. AB - We report the characterization of an integrated Ag+/Na+ ion exchange waveguide realized in a silicate glass substrate using apertureless scanning near-field optical microscopy. Our experimental set-up is based on the combination of a commercial atomic force microscope with an optical confocal detection system. Thanks to this system, the topography and evanescent optical field at the waveguide top surface are mapped simultaneously. Also, the process of apertureless scanning near-field optical microscopy image formation is analysed. In particular, fringe patterns appearing in the image reveal the intrinsic interferometric nature of the collected signal, due to interference between the field scattered by the tip end and background fields related to guide losses. We give a quantitative interpretation of these fringes. Evanescent intensity mapping on the sample surface allowed us to extract physical waveguide parameters. In particular, it shows an unambiguous multimode beat along the waveguide propagation axis. Furthermore, we show that analysis of this intensity profile reveals back-reflection effects from the waveguide exit facet. The resulting standing waves pattern allows us to evaluate the eigenmode propagation constants. PMID- 12641753 TI - Fabrication of near-field optical apertures in aluminium by a highly selective corrosion process in the evanescent field. AB - A simple, one-step process to fabricate high-quality apertures for scanning near field optical microscope probes based on aluminium-coated silicon nitride cantilevers is presented. A thin evanescent optical field at a glass-water interface was used to heat the aluminium at the tip apex due to light absorption. The heat induced a breakdown of the passivating oxide layer and local corrosion of the metal, which selectively exposed the front-most part of the probe tip from the aluminium. Apertures with a protruding silicon nitride tip up to 72 nm in height were fabricated. The height of the protrusion was controlled by the extent of the evanescent field, whereas the diameter depended on the geometry of the probe substrate. The corrosion process proved to be self-terminating, yielding highly reproducible tip heights. Near-field optical resolution in a transmission mode of 85 nm was demonstrated. PMID- 12641755 TI - Near-field and confocal surface-enhanced resonance Raman spectroscopy at cryogenic temperatures. AB - For laser spectroscopy at variable temperatures with high spatial resolution a combined scanning near-field optical and confocal microscope was developed. Rhodamine 6G (R6G) dye molecules dispersed on silver nano-particles or nano clusters were investigated. For optical excitation of the molecules, either an aperture probe or a focused laser spot in confocal arrangement were employed. Raman spectra in the wavenumber range between 300 cm-1 and 3000 cm-1 at room temperatures down to 8.5 K were recorded. Many of the observed Raman lines can be associated with the structure of the adsorbed molecule. Intensity fluctuations in spectral sequences were observed down to 77 K and are indicative of single molecule sensitivity. PMID- 12641756 TI - Surface plasmon micro- and nano-optics. AB - We report the experimental realization of key elements of a two-dimensional optics based on surface plasmon polaritons (SPPs). Local SPP sources, Bragg mirrors and beamsplitters were lithographically built from silver nanostructures. The SPP fields were imaged by detecting the fluorescence of molecules that act as local probes of the SPP near-field intensity. PMID- 12641757 TI - Near-field measurement of short-range correlation in optical waves transmitted through random media. AB - Two-dimensional near-field images of speckle patterns formed by optical waves transmitted through a disordered porous silica glass sample are measured. The corresponding 2D intensity correlation function, C, is extracted. The subwavelength spatial resolution of near-field microscopy allows us to resolve in the spatial distribution of C the expected subwavelength oscillations and to follow their dependence on the excitation wavelength. Finally, we deduce the effective refractive index of the material by fitting the theoretical spatial dependence of C to our experimental results. PMID- 12641759 TI - Characterization and fabrication of fully metal-coated scanning near-field optical microscopy SiO2 tips. AB - The fabrication of silicon cantilever-based scanning near-field optical microscope probes with fully aluminium-coated quartz tips was optimized to increase production yield. Different cantilever designs for dynamic- and contact mode force feedback were implemented. Light transmission through the tips was investigated experimentally in terms of the metal coating and the tip cone-angle. We found that transmittance varies with the skin depth of the metal coating and is inverse to the cone angle, meaning that slender tips showed higher transmission. Near-field optical images of individual fluorescing molecules showed a resolution < 100 nm. Scanning electron microscopy images of tips before and after scanning near-field optical microscope imaging, and transmission electron microscopy analysis of tips before and after illumination, together with measurements performed with a miniaturized thermocouple showed no evidence of mechanical defect or orifice formation by thermal effects. PMID- 12641760 TI - Phase separation in polyfluorene-polymethylmethacrylate blends studied using UV near-field microscopy. AB - In this paper we present a near-field microscopy study of thin films of a phase separated blend of the fluorescent conjugated-polymer poly(9,9-dioctylfluorene) [PFO] with the non-fluorescent polymer polymethylmethacrylate [PMMA]. A scanning near-field optical microscope (NSOM) was used to generate (blue) fluorescence from the PFO following UV excitation at 362 nm. A range of different concentrations of PFO in PMMA were studied ranging from 1 to 50% PFO in PMMA by mass. By studying both the shear force and fluorescence images we were able accurately to determine the distribution of PFO in the PMMA. We found that phase separation occurs over a number of different length-scales between 5 micro m and 250 nm. We show that at PFO concentrations of 1%, the PFO lies on top of the PMMA. At a PFO relative concentration of 50%, the PMMA phase extends through the whole thickness of the film to the underlying substrate. We use such samples to discuss the resolution of NSOM when imaging thick organic films. Furthermore, we confirm that the length-scales of phase separation can be modified via control over spin-casting protocols. PMID- 12641758 TI - Two-photon excitation of excitons in CuCl in total reflection geometry. AB - We have observed emission spectra of a CuCl film on a TiO2 prism surface(110) associated with the two-photon excitation of the exciton system in total reflection geometry. The I1 bound exciton emission, which resonantly appeared at the two-photon excitation of the Z3-longitudinal excitons and Z1,2-exciton band, was observed. The dependence of the emission intensities on the polarization of the excitation light was explained from the field intensity and the penetration depth of the evanescent light in CuCl accompanying the totally reflected light at the TiO2/CuCl interface. PMID- 12641761 TI - Near-field imaging of ultrathin magnetic films with in-plane magnetization. AB - A new approach to near-field magneto-optical imaging was developed capable of visualization of in-plane magnetization of ultrathin magnetic structures. The approach relies on the magneto-optical effect specific for thin magnetic layers and employs near-field transmission measurements of longitudinal and/or transverse magneto-optical effect arising from the presence of thin film interfaces. The near-field magneto-optical contrast of in-plane domain structure of ultrathin Co film has been demonstrated in different polarization configurations. PMID- 12641762 TI - Low-temperature scanning system for near- and far-field optical investigations. AB - A combined system for far- and near-field optical spectroscopy consisting of a compact scanning near-field optical microscope and a dedicated spectrometer was realized. The set-up allows the optical investigation of samples at temperatures from 10 to 300 K. The sample positioning range is as large as 5 x 5 x 5 mm3 and the spatial resolution is in the range of 1.5 micro m in the far-field optical microscopy mode at low temperatures. In the scanning near-field optical microscope mode the resolution is defined by the microfabricated cantilever probe, which is placed in the focus of a double-mirror objective. The tip-to sample distance in the scanning near-field optical microscope is controlled by a beam deflection system in dynamic scanning force microscopy mode. After a description of the apparatus, scanning force topography images of self-assembled InAs quantum dots on a GaAs substrate with a density of less than one dot per square micrometre are shown, followed by the first spectroscopic investigations of such a sample. The presented results demonstrate the potential of the system. PMID- 12641763 TI - Implementation of a short-tip tapping-mode tuning fork near-field scanning optical microscope. AB - We present the implementation of a short-tip tapping-mode tuning fork near-field scanning optical microscope. Tapping frequency dependences of the piezoelectric signal amplitudes for a bare tuning fork fixed on the ceramic plate, a short-tip tapping-mode tuning fork scheme and an ordinary tapping-mode tuning fork configuration with an 80-cm optical fibre attached are demonstrated and compared. Our experimental results show that this new short-tip tapping-mode tuning fork scheme provides a stable and high Q factor at the tapping frequency of the tuning fork and will be very helpful when long optical fibre probes have to be used in an experiment. Both collection and excitation modes of short-tip tapping-mode tuning fork near-field scanning optical microscope are applied to study the near field optical properties of a single-mode telecommunication optical fibre and a green InGaN/GaN multiquantum well light-emitting diode. PMID- 12641765 TI - Nano-patterning photosensitive polymers using local field enhancement at the end of apertureless SNOM tips. AB - We show experimentally that local optical field enhancement can occur at the end of an apertureless SNOM tip illuminated by an external light source. Our approach consists in the use of a photosensitive polymer, placed in the tip near-field, to record intensity distribution in the vicinity of the tip end. The excited nanometre-size light source permits us to produce nano-patterns on the polymer surface which are then characterized by atomic force microscopy. Experimental images show the influence, on the field enhancement, of three important experimental parameters: the polarization state of the incident light, the geometry of the external illumination and the radius of curvature of the tip apex. These results are shown to be in good agreement with two-dimensional numerical calculations based on the finite-difference time-domain method. We show preliminary nanometre-size patterns created by this nano-source excited at a metallic tip extremity and discuss the potential of this approach for near-field optical lithography. PMID- 12641766 TI - Tracking of secretory vesicles of PC12 cells by total internal reflection fluorescence microscopy. AB - Total internal reflection fluorescence microscopy is used to detect cellular events near the plasma membrane. Behaviours of secretory vesicles near the cell surface of living PC12 cells, a neuroendocrine cell line, are studied. The secretory vesicles are labelled by over-expression of enhanced green fluorescent protein-tagged Rab3A, one of the small G proteins involved in the fusion of secretory vesicles to plasma membrane in PC12 cells. Images acquired by a fast cooled charge-coupled device camera using conventional fluorescence microscopy and total internal reflection fluorescence microscopy are compared and analysed. Within the small evanescent range (< 200 nm), the movements of the secretory vesicles of PC12 cells before and after stimulation by high K+ are examined. The movements of one vesicle relative to another already docked on the membrane are detected. Total internal reflection fluorescence microscopy provides a novel optical method to trace and analyse the exocytotic events and vesicle specifically near a cell membrane without interference of signals from other parts of the cell. PMID- 12641767 TI - Near-field Raman imaging of morphological and chemical defects in organic crystals with subdiffraction resolution. AB - In this study we report on the application of an aperture near-field optical microscope for Raman imaging of organic materials. Spectral analysis and detailed Raman imaging are performed with integration times of 100 ms per point, without the aid of field enhancement effects. The studied samples consist of two high Raman-efficiency molecular samples: a 7,7',8,8'-tetracyanoquinodimethane crystal showing surface defects and a 7,7',8,8'-tetracyanoquinodimethane thin film characterized by the presence of submicrometre-sized organometallic copper-salt complexes. In the first case, the effect of the surface deformation was studied, whereas in the second sample we were able to chemically image the formation of salt complexes. Subdiffraction resolution was achieved in both studies. PMID- 12641764 TI - Surface plasmon polariton waveguiding in random surface nanostructures. AB - In this study, guiding of surface plasmon polaritons excited at a gold film surface along corrugation-free channels in regions that are covered with randomly located surface scatterers, is considered using near-field microscopy for imaging of surface plasmon polariton intensity distributions at the surface. In the wavelength range 713-815 nm, we observed complete inhibition of the surface plasmon polariton propagation inside the random structures composed of individual ( approximately 70 nm high) gold bumps (and their clusters) placed on a 55 nm thick gold film with a bump density of 75 micro m-2. We demonstrate well-defined surface plasmon polariton guiding along corrugation-free 2 micro m wide channels in random structures and, in the wavelength range 738-774 nm, low-loss guiding around 20 degrees bends having a bend radius of approximately 15 micro m. PMID- 12641768 TI - Near-field imaging of surface plasmon on gold nano-dots fabricated by scanning probe lithography. AB - We obtained scanning near-field optical microscopy images to study the excitation of surface plasmons on metallic dots fabricated using scanning probe lithography. Gold nano-dots were fabricated by applying electric voltages to conducting probes installed in an atomic force microscope using the mechanism of field-induced diffusion and nano-oxidation plus Au-coating. High spatial resolution of scanning near-field optical microscopy revealed a 'bifold' pattern of surface plasmon mode on fabricated Au dots in the polarization direction of incident light. We found that scanning near-field optical microscopy imaging combined with scanning probe lithography is able to provide a systematic study of surface plasmon excitation on nano-metallic structures. PMID- 12641770 TI - Single-molecule near-field optical energy transfer microscopy with dielectric tips. AB - The fluorescence lifetime and the fluorescence rate of single molecules are recorded as a function of the position of a Si3N4 atomic force microscopy tip with respect to the molecule. We observe a decrease of the excited state lifetime and the fluorescence rate when the tip apex is in close proximity to the molecule. These effects are attributed to the fact that the dielectric tip converts non-propagating near-fields to propagating fields within the dielectric tip effectively quenching the fluorescence. The spatial extension of the quenching area is of subwavelength dimensions. The results are discussed in terms of molecular fluorescence in a system of stratified media. The experiment provides surprising new insights into the interactions between a fluorescent molecule and a dielectric tip. The methodology holds promise for applications in ultra high-resolution near-field optical imaging at the level of single fluorophores. PMID- 12641769 TI - Photonic nanopatterns of gold nanostructures indicate the excitation of surface plasmon modes of a wavelength of 50-100 nm by scanning near-field optical microscopy. AB - Scanning near-field optical microscopy images of metal nanostructures taken with the tetrahedral tip (T-tip) show a distribution of dark and bright spots at distances in the order of 25-50 nm. The images are interpreted as photonic nanopatterns defined as calculated scanning near-field optical microscopy images using a dipole serving as a light-emitting scanning near-field optical microscopy probe. Changing from a positive to a negative value of the dielectric function of a sample leads to the partition of one spot into several spots in the photonic nanopatterns, indicating the excitation of surface plasmons of a wavelength in the order of 50-100 nm in metal nanostructures. PMID- 12641771 TI - Optical transition process in AgOx super-resolution near-field structure. AB - A sandwiched 15 nm AgOx thin film of the super-resolution, near-field optical disk was studied using a confocal Z-scan system. Nonlinear optical properties of quartz glass/ZnS-SiO2 (170 nm)/AgOx (15 nm)/ZnS-SiO2 (40 nm) were measured using a Q-switch Nd : YAG pulse laser of wavelength 532 nm, pulse width 0.7 ns, and 15.79 kHz repetition rate. Transmittance and reflectance of the sandwiched AgOx thin film show important optical responses at the focused position of Z-scan. The dissociation processes of AgOx, recombination of the silver and oxygen, and the resonance of the localized surface plasmon of the nano-composites of the AgOx thin film are correlated to transmittance and reflectance at the focused position of the Z-scan for different input laser powers. An irreversible upper threshold intensity of 4.40 x 106 mW cm-2 at the focused position was found. A reversible working window of the focusing intensity between 1.86 x 106 and 4.40 x 106 mW cm 2 was measured with sandwiched AgOx thin film alone. The near-field interactions of the AgOx thin film and the recording layers of super-resolution near-field optical disk are also discussed. PMID- 12641773 TI - Photoplastic near-field optical probe with sub-100 nm aperture made by replication from a nanomould. AB - Polymers have the ability to conform to surface contours down to a few nanometres. We studied the filling of transparent epoxy-type EPON SU-8 into nanoscale apertures made in a thin metal film as a new method for polymer/metal near-field optical structures. Mould replica processes combining silicon micromachining with the photo-curable SU-8 offer great potential for low-cost nanostructure fabrication. In addition to offering a route for mass production, the transparent pyramidal probes are expected to improve light transmission thanks to a wider geometry near the aperture. By combining silicon MEMS, mould geometry tuning by oxidation, anti-adhesion coating by self-assembled monolayer and mechanical release steps, we propose an advanced method for near-field optical probe fabrication. The major improvement is the possibility to fabricate nanoscale apertures directly on wafer scale during the microfabrication process and not on free-standing tips. Optical measurements were performed with the fabricated probes. The full width half maximum after a Gaussian fit of the intensity profile indicates a lateral optical resolution of approximately 60 nm. PMID- 12641772 TI - Demonstrating nanophotonic switching using near-field pump-probe photoluminescence spectroscopy of CuCl quantum cubes. AB - We demonstrated a novel optical switching operation using three CuCl quantum cubes with a size ratio of. Their quantized excitonic energy levels resonate with one another, and the switching mechanism was based on the resonant near-field energy transfer between the quantum cubes. Using near-field pump-probe photoluminescence spectroscopy, we succeeded in controlling the near-field energy transfer and obtained a controlled (i.e. switched) signal in a differential photoluminescence spectrum with and without a pump beam. The internal quantum efficiency of the switching operation was close to 1. These results suggest the possibility of making a nanophotonic switching device smaller than 30 nm. PMID- 12641775 TI - Cutaneous vasculitis: a review. AB - As the skin is commonly involved in systemic vasculitic disorders as well as those hypersensitivity states whose expression is largely skin-confined, cutaneous vasculitic lesions offer a window to diagnosis and a ready source of accessible tissue for biopsy. In this review, we discuss the pathologic manifestations of chronic vasculitic syndromes such as granuloma faciale and erythema elevatum diutinum; IgA-associated vasculitis including Henoch-Schonlein purpura; vasculitis seen in the setting of cryoglobulinemia and hypergammaglobulinemia of Waldenstrom, hereditary deficiencies of complement, and IgA deficiency; those leukocytoclastic vasculitides resulting from hypersensitivity reactions to drug, chemical and foodstuff ingestion; and those vasculitides seen in patients with systemic diseases such as polyarteritis nodosa, rheumatoid arthritis, mixed connective tissue disease, systemic lupus erythematosus, Sjogren's syndrome, relapsing polychondritis, Behcet's disease, Wegener's granulomatosis, and allergic granulomatosis of Churg and Strauss. PMID- 12641774 TI - Near-field optics and quantum optics: an assignation arranged by four kinds of photons. AB - In the process of emergence a photon emitted from an atom (or a molecule) is usually no better localized in space than to the near-field zone of the source. Near-field optics therefore is of central importance for understanding fundamental statistical aspects related to single-photon tunnelling, the space time description of photon dynamics, and the photon position-operator problem. In the present work an attempt is made to study the microscopic near-field optical interaction from a quantum statistical point of view. In near-field quantum electrodynamics (QED) scalar and longitudinal photons always are involved and this makes the covariant formulation of QED attractive also in the low-energy regime. We show that the Lorenz gauge condition on the global state vector relates to the near-field electrodynamics of the d-photons. The gauge photon is shown to be of no importance in near-field interactions. To understand the role of the lack of photon localizability we finally study near-field quantum optical correlations in a new so-called propagator gauge. PMID- 12641776 TI - Loss of expression of tuberin and hamartin in tuberous sclerosis complex associated but not in sporadic angiofibromas. AB - BACKGROUND: Angiofibromas occur sporadically, and they develop in most patients with tuberous sclerosis complex (TSC), which is associated with alterations of the tumor suppressor genes TSC1 or TSC2. Loss of tuberin, the protein product of TSC2, has been shown in the interstitial fibroblast compartment of TSC-associated angiofibromas. It is unclear whether there is also a loss of hamartin, the product of TSC1 in TSC-associated and sporadic angiofibromas. METHODS: The expression of hamartin and tuberin was analyzed by immunohistochemistry in 59 TSC associated and 12 sporadic angiofibromas using affinity-purified antibodies. RESULTS: Loss of expression of both tuberin and hamartin was detected in 14 angiofibromas, loss of only tuberin in three, and loss of only hamartin in four TSC-associated angiofibromas; but there was no loss in the sporadic angiofibromas. Only the interstitial cells, but not the vascular cells, showed a loss of expression of tuberin or hamartin. CONCLUSIONS: Loss of tuberin or hamartin occurred in a minority of the TSC-linked angiofibromas, but not in the sporadic angiofibromas. The absence of both tuberin and hamartin in some of the tumors suggests that the stability of tuberin and hamartin, which are believed to form an active complex in vivo, is negatively affected by the absence of either of the partners. PMID- 12641777 TI - Loss of mismatch repair proteins in sebaceous gland tumors. AB - BACKGROUND: Sebaceous gland neoplasms are rare tumors that are associated with visceral malignancies in patients with Muir-Torre syndrome (MTS). The majority of the MTS-associated tumors reveal mutations in DNA mismatch repair (MMR) genes (most often hMSH-2 and hMLH-1) and microsatellite instability. The sebaceous gland lesions in patients with MTS can often precede or occur concurrently with the visceral neoplasms. The early recognition of those lesions and their differentiation from sporadic sebaceous gland tumors are critical for proper patient management. Here we investigate the MMR gene expression in a variety of sebaceous gland tumors, with or without associated visceral malignancy. METHODS: We studied the expressions of hMLH-1 and hMSH-2 in 10 consecutive sebaceous hyperplasias, 10 sebaceus nevi, 12 sebaceous adenomas, seven sebaceous carcinomas and the adjacent normal sebaceous glands using immunohistochemistry and paraffin embedded sections. RESULTS: The normal sebaceous glands and the glands of all the sebaceus nevi were positive for hMLH-1 and hMSH-2. Loss of hMSH-2 expression was found in 1/10 (10%) sebaceous hyperplasias, 3/12 (25.0%) sebaceous adenomas, and 2/7 (28.6%) sebaceous carcinomas. Loss of hMLH-1 expression was seen in 1/10 (10%) hyperplasias, 3/12 (25.0%) adenomas, and 1/7 (14.3%) carcinomas. No concurrent loss of both hMLH-1 and hMSH-2 was observed. Loss of MMR (either hMLH 1 or hMSH-2) was detected in 80% of the benign sebaceous lesions associated with malignancy. In comparison, only 23% of sebaceous lesions not associated with malignancy showed loss of MMR proteins. No loss of hMSH-2 protein was found in the visceral cancer in one patient with hMSH-2-negative sebaceous adenoma. CONCLUSIONS: Our results confirm the previous reports of alterations of mismatch repair genes in the sebaceous neoplasms of patients with MTS. However, we showed that those changes also occur early at the stage of sebaceous hyperplasia, even in the absence of a visceral malignancy. This indicates the importance of the abnormal DNA mismatch repair in the progression of this disease. PMID- 12641778 TI - CD44 and hyaluronate in the differential diagnosis of dermatofibroma and dermatofibrosarcoma protuberans. AB - The histological distinction between dermatofibroma (DF) and dermatofibrosarcoma protuberans (DFSP) may be extremely difficult. CD34 and Factor XIIIa have been used to differentiate DF from DFSP. However, there is an overlap and relative lack of specificity of their expressions. CD44 is a widely distributed integral membrane glycoprotein, which is expressed as a multitude of isoforms generated by alternative splicing of at least 10 different variant exons and post translational modifications. CD44 is currently thought to be the principal cell surface receptor for hyaluronate (HA), the major component of the extracellular matrix. In this study we aimed to assess the expression of standard CD44 (CD44s) and its isoforms (CD44v3, CD44v4, CD44v5, CD44v6, CD44v7, CD44v7v8, and CD44v10), and HA in DF and DFSP. Immunohistochemical staining was performed on the biopsy specimens of 15 cases of DF and four cases of DFSP, using antibodies that recognize the CD44s, different CD44 isoforms and the hyaluronate binding protein (HABP). Tumor cells displayed a strong CD44s immunoreactivity in all cases of DF whereas a faint HA positivity was observed in the tumor stroma. The DF cells were negative for CD44v3, CD44v4, CD44v6, CD44v7 and CD44v7v8 but showed a strong reactivity for CD44v5 and CD44v10. In contrast, CD44s' expression was significantly reduced or absent in all DFSP lesions and the tumor stroma displayed strong staining for HA. Our results indicate that CD44 and HA can be used as additional diagnostic markers to distinguish DF from DFSP. PMID- 12641779 TI - Dermatofibrosarcoma protuberans of the vulva and groin: detection of COL1A1-PDGFB fusion transcripts by RT-PCR. AB - BACKGROUND: Dermatofibrosarcoma protuberans (DFSP) is an uncommon infiltrative tumor of the dermis with characteristic cytogenetic features, specifically the translocation t(17;22)(q22;q13) which often manifests as a supernumerary ring chromosome r(17;22). The translocation results in the fusion of two genes, collagen type I alpha 1(COL1A1) and platelet-derived growth factor B-chain (PDGFB). The trunk and extremities are the anatomic sites of predilection for DFSP, whereas the vulva and groin are quite uncommon sites of involvement. METHODS: This investigation evaluated seven DFSPs (four vulvar and three groin) for the presence of COL1A1-PDGFB fusion transcripts by reverse transcriptase polymerase chain reaction (RT-PCR), using archival formalin-fixed, paraffin embedded tissue. RESULTS: Six of seven cases (three vulvar, three groin) contained a COL1A1-PDGFB fusion transcript. Sequence analysis of the PCR products revealed that the break-point of the COL1A1 gene was located after exon 40 in two patients, after exon 42 in one patient, after exon 44 in one patient, and after exon 47 in two patients; the break-point in the PDGFB gene was before exon 2 in all cases. No fusion transcript could be amplified in one case. CONCLUSIONS: As in DFSP at other sites, COL1A1- PDGFB chimeric fusion transcripts are present in vulvar and groin DFSP. The transcripts can be detected in formalin-fixed, paraffin-embedded tumor tissues, and have the same general pattern of exon boundaries as in DFSP at other sites. PMID- 12641780 TI - Basal cell carcinoma with tricholemmal (at the lower portion) differentiation within seborrheic keratosis. AB - BACKGROUND: Recent genetic investigations support the idea that basal cell carcinoma (BCC) is trichoblastic carcinoma. However, it is generally thought that clear cell basal cell carcinoma is a result of degeneration rather than tricholemmal differentiation. METHODS: We report a case of BCC, with clear cell components, that developed within seborrheic keratosis, with histopathological and immunohistochemical findings. RESULTS: The clear cell components in the present case showed the following four characteristics: (i) at the periphery of the aggregations, columnar clear cells were aligned in a palisade along a well defined basement membrane; (ii) the nuclei of the columnar clear cells were at the pole opposite the basement membrane; (iii) the clear cells contained glycogen; (iv) in the aggregations with clear cell components, there was diffuse positive staining for cytokeratin 7 (CK7) (OV/TLR/30), but only the inner region stained positive for CK17. These four characteristics are comparable to those of the lower portion of normal outer root sheath. In addition, the BCC in the present case was partly composed of squamous cells that contained glycogen and were selectively positive for CK17 - features similar to those of squamous cells in normal outer root sheath. CONCLUSIONS: Some clear cell BCCs are simply the result of degenerative change, but other clear cell BCCs may be the result of tricholemmal (at the lower portion) differentiation. PMID- 12641781 TI - 'Eruptive syringoma': a misnomer for a reactive eccrine gland ductal proliferation? AB - BACKGROUND: Syringomas have traditionally been categorized as benign neoplasms of the eccrine gland ductal epithelium. However, the variety of clinical presentations reported in the literature and some cases recently observed by the authors cast doubt upon the neoplastic nature of eruptive syringomas. Our goal is to challenge the traditional notion that eruptive syringomas are neoplastic lesions. RESULTS: We observed two patients who presented with an eczematous process, which resolved leaving residual lesions. Biopsies of the late lesions showed features of eccrine syringoma. Yet a biopsy obtained from an incipient lesion in one of the cases showed a lymphocytic inflammatory reaction of the superficial portion of the eccrine duct resulting in tortuous hyperplastic changes. CONCLUSION: Based on our observations, some of the so-called 'eruptive syringoma' may represent a hyperplastic response of the eccrine duct to an inflammatory reaction rather than a true adnexal neoplasm. We proposed the term 'syringomatous dermatitis' for such cases. PMID- 12641783 TI - Carcinomatous transformation of eccrine syringofibroadenoma. AB - BACKGROUND: While squamous cell carcinoma and pseudocarcinomatous hyperplasia have been documented as pre-existing lesions in cases of reactive eccrine syringofibroadenoma (ESFA), to the best of our knowledge carcinoma occurring in a solitary ESFA has not yet been reported. We present one such case in a 91-year old female who had a dome-shaped, reddish tumor on the extensor side of the left forearm. METHODS: We review the histopathological, immunophenotypical and ultrastructural findings of this tumor, including the keratin expression profile. RESULTS: Histopathologically, long, branching, anastomosing, thin and thick strands of small cuboidal epithelial cells were extending from the surface epidermis into the dermis. In the center of the tumor, there were irregular shaped nests of atypical tumor cells invading downward into the dermis. Ultrastructurally, duct-like lumina lined with cuboidal tumor cells were present in the epithelial cords. From these findings, the present case was diagnosed as solitary eccrine syringofibroadenocarcinoma (ESFAC). Keratin expression studies revealed that cells of the thick strands, except for the luminal and basal cells, were positive for differentiation-specific keratins, keratins 1 and 10, and that cells of the thin strands were positive for keratins 5 and 14. CONCLUSIONS: Histopathological, immunophenotypical and ultrastructural evidence, as well as the pattern of keratin expression, suggest differentiation of the present malignant tumor towards the eccrine dermal duct. This case is the first reported case of ESFAC as far as we know. PMID- 12641782 TI - Mixed tubulopapillary hidradenoma and syringocystadenoma papilliferum occurring as a verrucous tumor. AB - Tubulopapillary hidradenoma (TPH)1 is a term proposed to describe morphological dermal ductal tumors with both eccrine and apocrine differentiation. The term TPH encompasses a spectrum of lesions that includes tubular apocrine adenoma (TAA) and papillary eccrine adenoma (PEA):2 PEA and TAA can be indistinguishable both clinically and histologically. We described a case of TPH with both prominent eccrine and apocrine differentiation combined with syringocystadenoma papilliferum (SCAP) over the distal extremity. This rarely encountered dermatopathological phenomenon is the sixth reported case from the literature in which PEA or TAA and SCAP were present in the same lesion.3-7 Furthermore, the tumor had a warty surface, which is histologically consistent with a typical viral verruca. Although PCR and DNA probe hybridization for human papilloma virus (HPV) types 2, 6/11, 16 and 18 failed to reveal positive results, the location and clinicopathologic correlation convinced us that superimposed HPV could not be excluded. PMID- 12641784 TI - HPV in verruciform xanthoma--sensitivity and specificity of detection methods and multiplicity of HPV types affect results. PMID- 12641787 TI - Pyoderma gangrenosum: a review. AB - Since its first description in 1930, the pathogenesis of pyoderma gangrenosum (PG) has remained obscure even as an ever-widening array of systemic diseases has been described in association with it. The histopathologic distinction of PG from other ulcerative processes with dermal neutrophilia is challenging and at times impossible. In consequence, when confronted with a biopsy from such a lesion, the pathologist has an obligation to obtain a full and detailed clinical history. In short, as a diagnosis of PG does not hinge exclusively upon the biopsy findings in isolation from other studies, a solid knowledge of the clinical features, the systemic disease associations and the differential diagnosis will help the pathologist to avoid diagnostic pitfalls or the generation of a report which is non-contributory to patient care. In this review, we describe in detail the different clinicopathologic forms of PG, summarize the diseases associated with this process in the literature and in our experience, and briefly review the treatment options. PMID- 12641788 TI - An immunohistochemical study of CD4, CD8, TIA-1 and CD56 subsets in inflammatory skin disease. AB - BACKGROUND: Antibodies to CD4, CD8, TIA-1, and CD56 are available which perform well in formalin-fixed and paraffin-embedded tissue. While previous studies have investigated CD4 and CD8 subsets in inflammatory skin disease, few have specifically addressed TIA-1 and CD56 reactivity in benign dermatoses. Given that CD8, TIA-1, and CD56 are linked to aggressive lymphoproliferative disorders (i.e. subcutaneous panniculitic T-cell lymphoma, natural killer (NK), and NK/T-cell lymphomas), it would be important to determine their specificity for cutaneous hematologic malignancies. This investigation was undertaken to determine the frequency with which common, benign dermatoses express these four markers. We also sought to determine whether the ratio of CD4- to CD8-positive cells could be used to distinguish among the dermatoses, especially the superficial and deep perivascular ones. METHODS: Formalin-fixed and paraffin-embedded sections from a variety of common inflammatory dermatoses were stained with antibodies to CD4, CD8, TIA-1, and CD56. Positive reactions were scored as a percentage of the entire mononuclear cell infiltrate. RESULTS: All of the dermatoses represented in the study showed TIA-1- and CD56-positive lymphocyte subpopulations. On a case-by case basis, the percentage of positive cells varied, and while all cases were positive for TIA-1, many were completely negative for CD56. For TIA-1, the percentage of positive cells ranged from 21 to 59%, and for CD56, from < 1 to 9%. The CD4:CD8 ratio ranged from 1.0 to 6.0 but was never less than 1.0. In addition to lymphocytes, TIA-1 also stained polymorphonuclear leukocytes, eosinophils, and mast cells. CONCLUSION: TIA-1- and CD56-positive lymphocytes are common participants in routine inflammatory dermatoses, and therefore these markers are not specific for aggressive lymphoproliferative disorders. Using only immunohistochemical data, the ratio of CD4- to CD8-positive lymphocytes could not be used reliably to separate the superficial and deep perivascular dermatoses from one another. Finally, mast cells are positive for TIA-1 and are commonly seen in normal and inflamed skin, and thus TIA-1 is not specific for cytotoxic T lymphocytes. PMID- 12641789 TI - Epithelioid sarcoma: an immunohistochemical analysis evaluating the utility of cytokeratin 5/6 in distinguishing superficial epithelioid sarcoma from spindled squamous cell carcinoma. AB - BACKGROUND: Epithelioid sarcoma (ES) is a rare, aggressive soft tissue tumor characterized by nodular aggregates of epithelioid and/or spindled cells that are immunoreactive to cytokeratins (CKs) and epithelial membrane antigen. ES that arises in the dermis may cause epidermal ulceration and can resemble, clinically, morphologically and immunohistochemically, cutaneous squamous cell carcinoma. CK 5/6 has recently been found to be an excellent marker of squamous cell carcinoma, including spindled variants, but it is not known if this marker can be utilized to distinguish superficial ES from cutaneous spindled squamous cell carcinoma (SSCC). METHODS: Twenty-four cases of ES with typical histologic features and 10 cases of SSCC with ultrastructural evidence of epithelial differentiation were studied. Immunohistochemical analysis using an antibody to CK 5/6 was performed. The extent of immunoreactivity was evaluated in a semiquantitative manner using the following scale: 0, < 5% of cells staining; 1+, 6-25% of cells staining; 2+, 26-50% of cells staining; 3+, 51-75% of cells staining; 4+, > 75% of cells staining. RESULTS: CK 5/6 was expressed in all 10 cases of SSCC, including one case with 3+ staining and six cases with 4+ staining. In contrast, CK 5/6 staining was found only in rare tumor cells (1+ staining) in one of 24 (4%) cases of ES. CONCLUSIONS: CK 5/6 staining is useful in distinguishing superficial ES from cutaneous SSCC. PMID- 12641790 TI - Antibody to S100a6 protein is a sensitive immunohistochemical marker for neurothekeoma. AB - BACKGROUND: Neurothekeoma is a benign tumor of putative peripheral nerve sheath origin. It occurs in a myxoid (classic) variant, cellular variant, and intermediate (mixed) variant. Cellular neurothekeoma (CNT) usually involves the head and neck or extremities of young patients. Histologically, CNT can be confused with melanocytic and fibrohistiocytic lesions. An immunohistochemical antibody panel is often necessary to confirm the histological impression and exclude melanocytic and/or fibrohistiocytic lesions. METHODS: Formalin-fixed, paraffin-embedded archival tissues were evaluated by immunohistochemistry using antibodies specific for S100A6 and PGP9.5 in 11 cases of neurothekeoma (seven cellular, four myxoid). A variety of other antibodies were evaluated by immunohistochemistry at the time of initial diagnosis. RESULTS: All 11 neurothekeoma cases were positive for S100A6 protein (four cases, weak/1+; seven cases, strong/2+), corresponding to 100% sensitivity. In contrast, eight of 11 neurothekeoma cases (73% sensitivity) were positive for PGP9.5. All seven CNT cases were negative for S100B, as expected. CONCLUSIONS: Anti-S100A6 is a highly sensitive antibody for neurothekeomas, including CNT, and, in our experience, is superior in sensitivity to PGP9.5. However, like other antibodies used in evaluating neurothekeomas, S100A6 lacks specificity, as has been demonstrated in previous studies. Nevertheless, S100A6 can be useful in an immunohistochemical antibody panel to evaluate lesions where the differential diagnosis includes CNT. PMID- 12641791 TI - Pathogenesis of solar elastosis: synthesis or degradation? PMID- 12641792 TI - Fibroblastic/myofibroblastic sarcoma of the skin: a report of five cases. AB - BACKGROUND: A number of malignant soft tissue tumors, particularly those of fibroblastic and fibrohistiocytic derivation, have been found to display myofibroblastic differentiation focally. The term myofibroblastic sarcoma, a controversial presumably distinctive entity, defines a malignant soft tissue tumor in which myofibroblasts are quantitatively the predominant cell type. METHODS: Five cases of cutaneous spindle-cell sarcomas showing fibroblastic myofibroblastic differentiation with predominance of fibroblasts were retrieved from the files of three large centers of dermatopathology. Tumors were analyzed histopathologically, immunophenotypically, and, in two cases, ultrastructurally. Results were compared with those previously reported in fibrosarcoma, malignant fibrous histiocytoma, and myofibroblastic sarcoma. RESULTS: Immunophenotypic and ultrastructural profiles of the cases analyzed in this series were closer to fibrosarcoma and to malignant fibrous histiocytoma than to myofibroblastic sarcoma by virtue of quantitative predominance of fibroblasts over myofibroblasts. On the other hand, histopathologic findings were in keeping with those reported in myofibroblastic sarcoma. CONCLUSIONS: Our series highlights the intrinsic problems in attaching certain cutaneous sarcomas with fibroblastic myofibroblastic differentiation to one of the recognized entities and gives support to the hypothesis that fibrosarcoma, malignant fibrous histiocytoma, and myofibroblastic sarcoma are related histogenetically. PMID- 12641793 TI - Effects of electromagnetic radiation from a cellular telephone on epidermal Merkel cells. AB - The number of reports on the effects induced by electromagnetic radiation (EMR) from cellular telephones in various cellular systems is still increasing. Until now, no satisfactory mechanism has been proposed to explain the biological effects of this radiation except a role suggested for mast cells. Merkel cells may also play a role in the mechanisms of biological effects of EMR. This study was undertaken to investigate the influence of EMR from a cellular telephone (900 MHz) on Merkel cells in rats. A group of rats was exposed to a cellular telephone in speech position for 30 min. Another group of rats was sham-exposed under the same environmental conditions for 30 min. Exposure led to significantly higher exocytotic activity in Merkel cells compared with the sham exposure group. This finding may indicate the possible role of Merkel cells in the pathophysiology of the effects of EMR. PMID- 12641794 TI - Effects of biopsy-induced wound healing on residual basal cell and squamous cell carcinomas: rate of tumor regression in excisional specimens. AB - BACKGROUND: Wound healing following a partial biopsy of basal cell (BCC) and squamous cell carcinomas (SCC) may induce tumor regression. METHODS: Nonmelanoma skin cancer (NMSC) biopsy and re-excision specimens from 1994 to 2001 were reviewed for histologic evidence of scar vs. presence of residual tumor in excision specimens. Regressed and non-regressed tumors were analyzed to assess the influence of anatomic location, biopsy technique (punch vs. shave), histologic subtype of BCC or SCC, time interval between biopsy and excision, and patient age. RESULTS: Nine hundred and ten excisions were performed for transected BCC or SCC, 217 (24%) of which showed scar with no residual tumor. Logistic regression analysis revealed significant differences in the regressed vs. non-regressed subsets. SCCs were more likely to regress than BCCs (40% vs. 20%, respectively, p < 0.00001). Independent of the NMSC type, tumors regressed more often following shave rather than punch biopsy (34% vs. 15%, respectively, p < 0.00001), as did tumors on the trunk and extremities compared with head and neck cases (31% vs. 21%, respectively, p < 0.01). CONCLUSIONS: In our series, 24% of NMSCs transected on the initial biopsy showed no residual tumor in the excision specimens, implying that some event in the interval between biopsy and excision may lead to the eradication of residual tumor. The exact mechanism is unclear, but wound healing likely plays an important role. PMID- 12641795 TI - Bednar tumor associated with dermal melanocytosis: melanocytic colonization or neuroectodermal multidirectional differentiation? AB - BACKGROUND: Neuroectodermal differentiation or melanocytic colonization are the opposing theories of histogenesis for the Bednar tumor or pigmented dermatofibrosarcoma protuberans (DFSP). OBSERVATION: A 31-year-old African American woman presented with a 2-cm blue-black shoulder nodule of 1-year duration. Punch biopsy revealed a CD34+, Factor XIIIa-DFSP, harboring numerous, pigmented spindle S100+, Mart-1+ and HMB-45+ cells. Subsequent wide excision demonstrated pigmented dendritic and spindled cells widely scattered throughout the dermis of the 3-cm excisional margins and punch biopsy specimens of normal skin from both shoulders. This latter process was interpreted as dermal melanocytosis (nevus of Ito). The dermal pigmented spindle cells were Mart-1+ and CD34-, and were associated with non-pigmented CD34+, cytologically banal spindle cells, which were more numerous in the excisional margins than the contralateral shoulder. CONCLUSION: Reported herein is a singular case of Bednar tumor associated with dermal melanocytosis. Although the coexistence of these processes implicates colonization of the DFSP by constituent dermal melanocytes, the mixed immunophenotype (CD34+ or Mart-1+ cells) of dispersed dermal spindle cells hints at the possibility of a common cell of origin: the putative neuromesenchymal cell. In effect, the Bednar tumor could represent one part of a spectrum of neural crest-derived dermal tumors that includes dermal melanocytosis, cellular blue nevus and conventional DFSP. PMID- 12641796 TI - Epithelioid angiosarcoma of the breast involving the skin: a highly aggressive neoplasm readily mistaken for mammary carcinoma. AB - BACKGROUND: Angiosarcomas are malignant neoplasms of endothelial cells. Angiosarcoma of the breast is a rare neoplasm that behaves in a highly malignant fashion. It must be differentiated from benign vascular proliferations and from mammary carcinoma. METHODS: We report on a 49-year-old-woman who presented with a large mass involving the left breast. RESULTS: The lesion had an erythematoviolaceous hue and livedoid pattern at the periphery. Histopathologic study showed an epithelioid malignant neoplasm, and immunohistochemical studies demonstrated that neoplastic cells expressed immunoreactivity for endothelial cell markers. CONCLUSIONS: A diagnosis of epithelioid angiosarcoma of the breast was established. The patient was treated with radical mastectomy, but she refused any other additional therapy. PMID- 12641797 TI - Reporting tumor thickness for cutaneous squamous cell carcinoma. PMID- 12641807 TI - The salience of the body: transition in sexual self-identity for women living with multiple sclerosis. AB - AIM: The purpose of this paper is to outline understandings about the construction of sexuality and the impact of a changing body for women living with multiple sclerosis (MS). We suggest that the process of transition towards incorporating the experience of chronic illness into one's life is influenced by the (re)construction of self-identity. DESIGN AND METHODS: A participatory action process guided the research. The women joined the authors for five group sessions that totalled 15 hours of contact time. In addition, we offered women the opportunity for one-to-one interviews at home. Nine women volunteered to participate. This allowed us to gain additional in-depth data about individual experiences. The interpretive framework was guided by the self-identity literature. When reading the transcripts we questioned: What is going on here? What does this say about the construction of self? What does this say about the construction of identity? What influence does the body have in the construction of self-identity? Analysis was collaborative (with the women) and the resultant emerging construction of sexuality is shared in this paper. Data generated during one-to-one interviews are privileged and we include two accounts from women who live with MS. The women's stories focus on sexuality, however, within this sexual context, we observed shifts in self-identity which we contend may shape the illness transition experience. FINDINGS: The rationale for privileging only two accounts is to expand understanding of Ordinariness and Extraordinariness with particular focus on the salience of the body in the 'sexual' lives of the women. Self-identity was shaped by how they felt about themselves as sexual beings, how they experienced their body, how they felt about sexual activities and by the way others reacted to them. Importantly, we view the women's sense of self, identity and the relationship to the body and find that shifts in self identity shape the woman's transition towards Ordinariness. CONCLUSIONS: This exploration of illness experiences is a reminder that our bodies are vehicles for our sense of self and identity. Cultural, educational, social, religious and family contexts all impact on women's capacity to shape the consequences of illness and the choices available to them. Facilitating women towards an awareness of the choices available in order to sustain or reclaim self may in turn expedite transition towards Ordinariness so that illness may become a part of their life. PMID- 12641808 TI - Postnatal quality of life: a pilot study using the Mother-Generated Index. AB - BACKGROUND: Although postnatal morbidity has been well documented in recent years, postnatal quality of life has not been addressed. A newly derived subjective measurement of postnatal quality of life (the Mother-Generated Index) combines a quantitative and qualitative evaluation. AIMS: This part of our pilot study aimed to compare the aspects of their lives nominated by women with low and high quality of life (Primary Index) scores, and to examine the respective importance of these areas. METHODS: The Mother-Generated Index was tested using the Edinburgh Postnatal Depression Scale, Short Form 12, and an established maternal and neonatal physical morbidity index as validators. Four health visitors administered these at 6-8 weeks and 8 months postpartum to 103 women by structured face-to-face interviews between June 2000 and March 2001. Data were entered into Epi-Info, and exported to Microsoft Excel and SPSS for analysis. RESULTS: A wide variety of quality of life aspects were reported, including emotional, social and financial concerns. Tiredness was prevalent in all groups, but other physical problems were rare at 8 months. Mothers with low quality of life (Primary Index) scores at 6-8 weeks and 8 months commonly reported having less personal time. Low scoring areas, which health professionals might consider in greatest need of attention, were often not the ones mothers deemed most important. LIMITATIONS: The study involved only 103 participants, and did not assess the degree of support experienced by the mothers. CONCLUSIONS: The Mother Generated Index helps mothers to identify the areas of their lives which are of most concern to them. This pilot suggests that mothers with high and low quality of life scores have markedly divergent experiences. PMID- 12641809 TI - Effectiveness of informational support in reducing the severity of postnatal depression in Taiwan. AB - AIM: The aim of the study was to investigate the effectiveness of informational support in reducing the Edinburgh Postnatal Depression Scale (EPDS) scores in Taiwan. METHODS: Five hundred Taiwanese women were screened during the fourth week after giving birth and those with a score of over 10 on the EPDS were considered to be at risk of postnatal depression. Seventy postpartum women were randomly allocated to the experimental or control group. Only those in the experimental group received informational support about postnatal depression during the sixth week postpartum. Two groups were assessed by the EPDS at 3 months postpartum to explore their depressive status. RESULTS: Taiwanese women who received informational support about postnatal depression 6 weeks after giving birth experienced lower EPDS scores at 3 months postpartum than those who did not receive this information. CONCLUSIONS: The findings suggest that informational support about postnatal depression given to women in the postnatal period may contribute to psychological well-being. PMID- 12641810 TI - Interaction between caregivers and families expecting a malformed child. AB - BACKGROUND: Earlier research has shown that the care of families expecting a malformed child should be intensified and that the topic is understudied. AIM: This study aimed at generating a practical nursing theory of interaction between caregivers and families expecting a malformed child, as experienced by families. METHODS: A grounded theory study was undertaken at a university hospital, Finland. Data consisted of semi-structured interviews with 29 mothers and fathers, analysed using the constant comparative method. FINDINGS: The interaction process starts with the confirmation of the diagnosis. It usually comes as a shock to the parents, and gives rise to questions about the nature of the malformation, family members' relationship with themselves, family relationships and relationships with people outside the family. These questions are crucial to the family's coping and form the core of interaction. Families' own coping strategies are either reinforced or undermined by caregivers' actions and are reflected in parents' expectations about caregivers, which in part account for their actions and experiences of interaction. The care system as an interactive environment is part of a process which results in the experiences of being helped or being left without help. The core of the interaction process comprises two dimensions: gaining strength and losing strength in relation to malformation issues. CONCLUSIONS: The findings of the study are consistent with earlier research and complement it by providing a detailed delineation of the interaction from the perspective of the family. They can be used to formulate recommendations for improving caregivers' interactive skills and awareness of the topic through training. The care system should be developed to ensure that families receive psychological support and that the family as a whole receives proper care. Consideration of the viewpoint of siblings is a challenge for further research. PMID- 12641811 TI - Self-respect, dignity and confidence: conceptions of integrity among male patients. AB - BACKGROUND: Integrity is a complex and important concept in ethical reasoning and in nursing care. Integrity is part of being a human being, a wholeness, and in this sense it also refers to health. Preserving patients' integrity is an important aspect of nursing care, as a number of situations arise in which they can feel threatened or violated. Integrity can also refer to acting according to general ethical principles and rules, i.e. moral integrity. AIM: The aim of this Swedish study was to describe how male patients conceive integrity. METHODS: Seventeen male patients were strategically selected for interviews, which were analysed by a phenomenographical approach. FINDINGS: The patients expressed 12 different conceptions of integrity that were summarized into three description categories: self-respect, dignity and confidence. This means that patients give themselves integrity when they believe in themselves, have the courage to set boundaries, have control over themselves and their situation, and have the possibility of being alone. Caregivers can preserve a patient's integrity by seeing him as trustworthy and as a whole person, and by showing respect. Confidence in a caring situation is characterized by trust in professionals, a balance between patients' and the caregivers' wishes, patient participation, the possibility of being free and confidentiality of information. DISCUSSION: Integrity could be described as what something 'is' and as a 'relationship' with oneself and others. It is imperative that caregivers are aware of patients' conceptions of integrity in order to identify and preserve their integrity and also so that they treat them in accordance with moral integrity. The findings are limited to male patients, and questions about female patients' conceptions are raised. PMID- 12641812 TI - Cognitive interviewing: verbal data in the design and pretesting of questionnaires. AB - PURPOSE: The purpose of this paper is to discuss problems that occur in questionnaire responses and how cognitive interviewing can be used to identify problematic questions prior to using the questionnaire in the field. BACKGROUND: Questionnaire design involves developing wording that is clear, unambiguous and permits respondents successfully to answer the question that is asked. However, a number of problems in relation to respondents' understanding and successfully completing questionnaires have been identified. Cognitive interviewing, an amalgamation of cognitive psychology and survey methodology, has been developed to identify problematic questions that may elicit response error. The overall aim is to use cognitive theory to understand how respondents perceive and interpret questions and to identify potential problems that may arise in prospective survey questionnaires. METHODS: A literature review is used to examine the process of questionnaire design and how cognitive interviewing can be used to reduce sampling error and increase questionnaire response rates. FINDINGS: Cognitive interviewing involves interviewers asking survey respondents to think out loud as they go through a survey questionnaire and tell them everything they are thinking. This allows understanding of the questionnaire from the respondents' perspective rather than that of the researchers. Cognitive interviews have been used in a number of areas in health care research to pretest and validate questionnaires and to ensure high response rates. Interviewing has been found to be highly effective in developing questionnaires for age specific groups (children and adolescents) and in ascertaining respondents' understanding in health surveys prior to distribution. However, cognitive interviews have been criticized for being overly subjective and artificial. CONCLUSION: Cognitive interviews are a positive addition to current methods of pretesting questionnaires prior to distribution to the sample. They are most valuable in pretesting questions that are complex, where questions are sensitive and intrusive and for specific groups for whom questionnaire completion may pose particular difficulties. PMID- 12641814 TI - The Internet and access to evidence: how are nurses positioned? AB - BACKGROUND: Published literature that describes the use of the Internet by nurses is scant, but it does reveal that there has been a delay in the acceptance of the Internet as a workplace tool by the medical community and, in particular by nurses. AIMS: The purpose of this article is to report on a study of how often and from what location nurses accessed the Internet, as well as the types of information they were seeking. In addition, our goal was to compare nurses' Internet use with that of physicians and the public at large, and to highlight structural and institutional challenges to nurses' use. METHODS: Surveys (1996 and 1998) of Alberta Registered Nurses were used to examine their use of technology at work and at home. Additional data sources were used to compare nurses to physicians and to the general public. RESULTS: While nurses' Internet and e-mail use at home increased over the 2-year period and was comparable with other groups, Internet use at work was low compared with other groups despite adequate workplace access. CONCLUSIONS: Nurses are more likely to value interpersonal contact, and prefer to use personal experience and communication with colleagues and patients rather than on-line and traditional sources of practice knowledge. In order for an information source to be seen as valuable in the clinical setting, contextually relevant information needs to be accessed quickly and efficiently. Energies should be focused on constructing information systems that address the particular needs of nurses. PMID- 12641813 TI - Comparing doctor- and nurse-led care in a sexual health clinic: patient satisfaction questionnaire. AB - BACKGROUND: A new model of comprehensive care nurse-led clinics has enabled experienced genitourinary medicine nurses to co-ordinate the first-line, comprehensive care of female patients presenting with sexually transmitted infections and other sexual health conditions and issues. AIM: This paper describes the development of a patient satisfaction questionnaire to compare the satisfaction of women attending nurse-led or doctor-led clinics at a central London genitourinary medicine clinic. METHODS: A previously validated questionnaire was adapted using the findings of qualitative interviews exploring patient expectations of the service. The draft questionnaire was tested for internal consistency, sub-scale homogeneity, construct validity and stability. The final version consisted of a 34 item, five-point Likert scale, which was found to be both reliable (Cronbach's alpha 0.91) and stable (test-retest 0.95). There was some evidence of construct validity. The questionnaire was then distributed to a convenience sample of 132 women attending a nurse-led clinic and 150 seen at a doctor-led clinic. RESULTS: There was a 90% response rate. The median total satisfaction scores, out of a total of five, were 4.47 and 4.30 for the nurse-led and doctor-led groups, respectively (P = 0.05). Significantly higher scores on the sub-scales measuring quality and competence of technical care (P < 0.001), provision of information (P = 0.01) and overall satisfaction (P = 0.01) were seen for the nurse-led group. No significant differences were found in the sub-scales measuring service attributes and specific attributes of interpersonal relationships. CONCLUSION: The rigorous development, piloting and testing phases of this satisfaction questionnaire led to reliable and valid results. This study demonstrated that nurse-led clinics within this service are an acceptable alternative to the existing doctor-led clinics. PMID- 12641815 TI - Nursing morale: what is it like and why? AB - BACKGROUND: There has been increased concern in the United Kingdom about the problems of recruitment and retention in the nursing profession, and how this is influenced by low morale. Despite this, however, there are relatively few studies, particularly of a qualitative nature, where nurses are asked about their morale and what factors affect it. AIMS: The present study aimed to explore nursing morale and determine the factors which nurses believed to influence it. By doing so, it was hoped that factors which affect nurse recruitment and retention could be identified. METHODS: Fifty-eight nurses (28 males, 30 females) working in the National Health Service in Scotland were interviewed in depth about their morale and their concerns about their career. Thematic analysis was carried out and a number of main issues emerged. FINDINGS: Morale in this group was very low. A large number of nurses were considering leaving the profession and the majority would discourage others from becoming a nurse. The themes that emerged, which related to their disillusionment, included low pay, lack of support for education, limited opportunity for promotion, lack of resources and job insecurity. CONCLUSIONS: The findings suggest that while recent salary increases may have helped to improve morale, other factors must also be addressed if further decline in morale and a subsequent nursing shortage is to be avoided. PMID- 12641816 TI - Horizontal violence: experiences of Registered Nurses in their first year of practice. AB - BACKGROUND: Interpersonal conflict among nurses (traditionally called 'horizontal violence' or 'bullying') is a significant issue confronting the nursing profession. However, there is a dearth of research focusing on horizontal violence experienced by new graduate nurses. AIMS: In order to assess the priority for preventive intervention programmes, the aims of this study were to determine the prevalence of horizontal violence experienced by nurses in their first year of practice; to describe the characteristics of the most distressing incidents experienced; to determine the consequences, and measure the psychological impact, of such events; and to determine the adequacy of training received to manage horizontal violence. METHOD: An anonymous survey was mailed to nurses in New Zealand who had registered in the year prior to November 2000 (n = 1169) and 551 completed questionnaires were returned (response rate 47%). Information was requested on the type and frequency of interpersonal conflict; a description of the most distressing event experienced; the consequences of the behaviour; and training to manage such events. The Impact of Event Scale was used to measure the level of distress experienced. RESULTS: Many new graduates experienced horizontal violence across all clinical settings. Absenteeism from work, the high number of respondents who considered leaving nursing, and scores on the Impact of Event Scale all indicated the serious impact of interpersonal conflict. Nearly half of the events described were not reported, only 12% of those who described a distressing incident received formal debriefing, and the majority of respondents had no training to manage the behaviour. CONCLUSIONS: First year of practice is an important confidence-building phase for nurses and yet many new graduates are exposed to horizontal violence, which may negatively impact on this process. The findings underscore a priority for the development of effective prevention programmes. Adequate reporting mechanisms and supportive services should also be readily available for those exposed to the behaviour. PMID- 12641817 TI - An in vitro examination of an extracellular matrix scaffold for use in wound healing. AB - This paper describes evidence that an extracellular matrix (ECM) secreted by human umbilical vein endothelial cells (HUVECs) assembled on gelatin coated plates overlaid by a mixed matrix secreted by human dermal microvascular endothelial cells (HDMECs) and human dermal fibroblasts provides a viable acellular scaffold for use in wound healing. Trypsinized epidermal keratinocytes or colonies from Dispase-digested fresh and cadaver skin tissue adhered and proliferated on either HUVECs ECM/gelatin or mixed matrix overlaid on HUVECs ECM/gelatin. An epithelial-mesenchymal interaction, previously thought to be tissue-specific, was exposed as well as concomitant integrin versatility. Furthermore, heterologous HDMECs and dermal fibroblasts attached and proliferated on the mixed matrix as well as HUVECs ECM. The conditioned medium from HUVECs (HUVECs CM) was found to neutralize the lingering after effects of Dispase, and could be used for the tissue culture of epidermal keratinocytes, HDMECs and dermal fibroblasts, which share related extracellular secretions. Taken together, these results indicate that cultured epithelial autografts can be redesigned to include both epithelial and dermal elements, and advances the acellular 'sandwich' ECM scaffold as a possible structural replacement for the lamina densa and lamina lucida, damaged or completely missing in some wounds and burns. PMID- 12641818 TI - CpG oligodeoxynucleotides accelerate reovirus type 2-triggered insulitis in DBA/1 suckling mice. AB - We reported previously that reovirus type-2 (Reo-2) triggers T-helper (Th) 1 mediated autoimmune insulitis resulting in temporal impaired glucose tolerance (IGT) approximately 10 days post infection (d.p.i) in suckling DBA/1 mice. We hypothesized that CpG motifs in bacteria may enhance virus-induced insulitis through its content of unmethylated CpG motifs. In the infected mice, the intraperitoneal treatment of synthetic 20-base oligodeoxynucleotides with CpG motifs (CpG ODN) caused increase in cumulative incidence of insulitis with IGT, increased serum interferon (IFN)-gamma concentration, and high frequency of autoantibody against pancreatic islet cells, compared to the infected mice without CpG ODN at 17 d.p.i. Also CD4+ and CD8+ lymphocytes infiltrated in and/or around pancreatic islets in the CpG ODN-treated mice. This evidence suggests that CpG ODN may contribute to accelerate Reo-2-induced autoimmune reaction against pancreatic islet cells via additional effects of Th1 cytokines especially IFN gamma. PMID- 12641819 TI - Studies on the haemotoxicity of chloramphenicol succinate in the Dunkin Hartley guinea pig. AB - In man, chloramphenicol (CAP), induces two major haemotoxic effects. First, a reversible, dose-related reticulocytopenia and anaemia developing during treatment. Second, a non-dose-related aplastic anaemia (AA), developing weeks after treatment, is often irreversible and fatal. In previous studies, we developed a mouse model of the reversible reticulocytopenia/anaemia using CAP succinate (CAPS); attempts to induce AA in the mouse with CAPS were unsuccessful; in the rat, CAPS induced only minimal haemotoxicity. We therefore wished to investigate haematological changes caused by CAPS in a third rodent, particularly in relation to the induction of significant 'late stage' bone marrow depression (AA). Female guinea pigs were gavaged with CAPS in three experiments. In a dose ranging study, CAPS (at 2500 and 3500 mg/kg) was administered daily for 9 days, and blood examined at 1 day post dosing. CAPS induced increased erythrocyte values (an apparent haemoconcentration effect), and reduced reticulocytes and femoral marrow nucleated cell counts (FNCC). In a second experiment, CAPS was given at 333, 666 and 1000 mg/kg (13 days); haematological changes were compared with results from the initial study, with evidence of dose-related effects. In a final experiment, CAPS was administered (825 mg/kg, 16 days) and blood studied at 1, 12, 28 and 63 days post dosing. At day 1, erythrocyte values were decreased (NS), and reticulocytes and FNCC were reduced; the marrow was hypocellular with erythroid depletion. At 12 and 28 days, values returned towards the normal range. At 63 days, parameters were normal. Thus, CAPS (825 mg/kg for 16 days) induced changes comparable to the reversible bone marrow depression seen in man; but there was no evidence of 'late stage' (i.e. at 63 days) marrow depression, as would be seen in a developing or overt marrow aplasia (AA). The guinea pig (like the mouse) is a model for the early events, but is not a good model for CAP induced AA in man. PMID- 12641820 TI - Serum and tissue level of insulin-like growth factor-I (IGF-I) and IGF-I binding proteins as an index of pancreatitis and pancreatic cancer. AB - Previously we have found deregulation of collagen metabolism in human pancreatitis and pancreatic cancer tissues. Insulin-like growth factor-I (IGF-I) is known to stimulate collagen biosynthesis through interaction with IGF-I receptor. IGF-I binding proteins (BPs) regulate the activity of IGF-I. We investigated whether serum and tissue IGF-I and IGF-BPs as well as tissue IGF-I receptor expression may reflect disturbances of collagen metabolism in patients with pancreatitis and pancreatic cancer. In pancreatitis tissue, a significant increase in IGF-I and IGFBP-3 content was accompanied by a distinct increase in IGF-I receptor expression, compared to control pancreas tissue. In contrast, serum from patients with pancreatitis did not show significant increases in IGF-I and IGFBP-3 levels, however, significant increases in IGFBP-1 level (2.5 fold). Moreover, a distinct decrease in radioactive IGF-binding to the BPs, compared to control serum, was found. Pancreatic cancer tissue and serum of patients with pancreatic cancer showed significant increases in IGF-I, IGFBP-3 and IGFBP-1 content, accompanied by dramatic increases in IGF-I tissue receptor expression, compared to controls. In serum of patients with pancreatic cancer distinct increases in radioactive IGF-binding to 46 kDa BP, compared to control serum, were observed. The data suggest that disturbances in tissue collagen metabolism during pancreatic diseases may result from deregulation of IGF-I homeostasis and that elevated serum levels of IGF-I, IGFBP-3 and IGFBP-1 may serve as markers of pancreatic cancer. PMID- 12641821 TI - A novel animal model of thymic tumour: development of epithelial thymoma in transgenic rats carrying human T lymphocyte virus type I pX gene. AB - The pX region encodes a major product of human T lymphocyte virus type I (HTLV-I) that has been implicated previously in tumour formation. To investigate the pathogenesis of pX gene in lymphoid tissues, we established a series of novel transgenic rats carrying the pX gene under the control of a rat lymphocyte specific protein tyrosine kinase (p56lck) proximal promoter. The transgene was constructed with the -269 to +26 region of a rat p56lck proximal promoter and the pX cDNA, and was microinjected into fertilized ova of Fischer 344/jcl female rats. Six transgenic lines from 114 pups were established. Integration and expression of the transgene were detected by polymerase chain reaction (PCR) and Southern hybridization or by reverse transcriptase-PCR, northern hybridization, and immunostaining. Thymic tumours with lethal expansion occurred in 4 of 6 transgenic lines. The tumour consisted of spindle shaped cells. Immunohistochemical and ultra-structural analysis characterized the tumour cells to as epithelial cell type, and in the tumour arose in the medulla. Therefore, the tumour is classified into predominantly epithelial and spindle cell of medullary thymoma (type A of the new World Health Organization classification), as based on the human classification. Tumor occurrence increased in proportion to levels of the pX transcription in the thymus, for each line, and sex distinction was evident regarding rates related to tumour expansion. The transgenic rat model described here is suitable as a model for analysing tumorigenesis in epithelial thymoma occurring in humans. PMID- 12641823 TI - ICSI: a technique too far? AB - Even now 10 years after its introduction, there is still debate on the possible adverse effects of intracytoplasmic sperm injection (ICSI). ICSI has raised a lot of concerns because of the mechanical perforation of the oocyte, the possible transmission of foreign genetic material, the use of immature or senescent germ cells and the association between genetic disorders and some forms of male infertility. To date, the data available indicate that ICSI is a safe procedure provided this treatment is performed in clinics with the highest standards of expertise and with a continuous follow-up programme for the offspring. It is becoming increasingly apparent that it is not ICSI itself, but the background risks of the ICSI patients that represent the major risk factor with regard to congenital malformations. More sex-chromosome abnormalities have been reported in ICSI offspring. Again, the risk of aneuploidy in ICSI progeny reflects the higher aneuploidy rate in the sperm of ICSI fathers. Whether ICSI will eventually perpetuate male infertility is far from clear, because at present the inheritance pattern of idiopathic male infertility is unknown. All ICSI candidates should nevertheless be thoroughly screened and counselled. As long as follow-up studies have a limited power to detect small increases in malformations and as long as no information is available on long-term and next-generation cohorts, ICSI must be used with caution and only when no alternative evidence-based therapy is available. All ICSI candidates should be rigorously screened and thoroughly informed of the limitations of current screening methods and our limited knowledge of the genetic background to male infertility. They should be told that there might be a slight increase in congenital malformation rate after ICSI, but that this increase is probably the result of their own 'background risk'. Well informed patients are in the best position to judge whether the concerns outweigh the benefits of ICSI. PMID- 12641822 TI - Effect of Helicobacter pylori infection on epidermal growth factor receptor (EGFR) expression and cell proliferation of gastric epithelial mucosa: correlation to macroscopic and microscopic diagnosis. AB - Our aim was to compare the expression of EGFR and proliferative cell nuclear antigen (PCNA) in different histological and endoscopic diagnostic groups, in cases of Helicobacter pylori infection, in vivo. Paraffin embedded human gastric biopsy samples (86) were analysed by EGFR and PCNA immunohistochemistry and classified both on the basis of histology and endoscopic findings. In normal epithelia (NE), a positive correlation was found between PCNA and EGFR and in H. pylori-negative gastritis with and without intestinal metaplasia (P < 0.01). On the other hand, a negative correlation was detected between the two immunohistochemical findings in H. pylori-associated gastritis with intestinal metaplasia (HPGIM) and in the atrophic gastritis (AG) group. In HPGIM the percentage of EGFR-positive cells was significantly lower (32.4 +/- 30.4) when compared to either the NE (50.3 +/- 23.7) or H. pylori-negative gastritis with intestinal metaplasia (HNGIM) (48.3 +/- 23.7). In AG, EGFR was significantly lower when compared to the NE (P < 0.05). Based on the endoscopic findings, a significant decrease of EGFR expression was found in gastric ulcer cases as compared to NE, gastritis or erosion cases (P < 0.01). PCNA showed no significant alterations between the NE and gastritis, AG groups. The presence of H. pylori has an inverse effect on PCNA and EGFR expression in HPGIM. PMID- 12641824 TI - The Y chromosome and male fertility and infertility. AB - Since 1995, thanks to a large number of studies, Y chromosome microdeletion screening has become part of the routine diagnostic work-up of severe male factor infertility. Many initial contradictory issues such as variability in deletion frequency, markers to be tested, presence of deletions in 'fertile' men, and genotype-phenotype correlation has been resolved. Past and present unresolved issues are discussed in this review. PMID- 12641826 TI - Evaluation of damage to the testicular cells of golden hamsters caused by experimental cryptorchidism using flow cytometry and confocal microscopy. AB - Artificial unilateral cryptorchidism was performed in golden hamsters which were then held for different periods of time. The non-operated side was used as a control. At various times from 4 to 15 days, hamsters were killed, testes were removed and weighed, single cell suspensions were prepared for flow cytometry analysis and seminiferous tubules were fixed for confocal microscopy. Using DNA staining by propidium iodide or acridine orange followed by flow cytometry analysis, a marked decrease in the haploid condensed cell fraction was detected at the beginning stages of experimental cryptorchidism. In correlation with flow cytometry results, spermiogenic arrest at stages IX and X of seminiferous epithelium was detected in these animals by confocal microscopy and there were no mature forms of haploid cells in the cryptorchid testis. In the testis with more severe damage, there were almost no haploid cells in the seminiferous tubules of cryptorchid animals. In addition, a significant decrease in tetraploid cell fraction and an increase in S-phase fraction was obtained in severe cases. This may be explained by cell arrest before entrance into meiosis. Destruction of tubule structure and cell arrangement were also observed by confocal microscopy in such cases. In conclusion, flow cytometry, combined with confocal analysis, added useful information about spermatogenesis disturbances in cryptorchid testis and it may be used as diagnostic tools in other cases of spermatogenic disorders. PMID- 12641825 TI - The CAG repeat polymorphism within the androgen receptor gene and maleness. AB - The androgen testosterone and its metabolite dihydrotestosterone exert their effects on gene expression and thus effect maleness via the androgen receptor (AR). A diverse range of clinical conditions starting with complete androgen insensitivity has been correlated with mutations in the AR. Subtle modulations of the transcriptional activity induced by the AR have also been observed and frequently assigned to a polyglutamine stretch of variable length within the N terminal domain of the receptor. This stretch is encoded by a variable number of CAG triplets in exon 1 of the AR gene located on the X chromosome. First observations of pathologically elongated AR CAG repeats in patients with X-linked spino-bulbar muscular atrophy showing marked hypoandrogenic traits were supplemented by partially conflicting findings of statistical significance also within the normal range of CAG repeat length: an involvement of prostate tissue, spermatogenesis, bone density, hair growth, cardiovascular risk factors and psychological factors has been demonstrated. The highly polymorphic nature of glutamine residues within the AR protein implies a subtle gradation of androgenicity among individuals within an environment of normal testosterone levels providing relevant ligand binding to ARs. This modulation of androgen effects may be small but continuously present during a man's lifetime and, hence, exerts effects that are measurable in many tissues as various degrees of androgenicity and represents a relevant effector of maleness. It remains to be elucidated whether these insights are important enough to become part of individually useful laboratory assessments. PMID- 12641827 TI - Identification of neurones of the brain and spinal cord involved in the innervation of the ductus deferens using the viral tracing method. AB - Using the viral transneuronal tracing technique cell groups of the spinal cord and brain transsynaptically connected with the ductus deferens were identified. Neurotropic (pseudorabies) virus was injected into the muscular coat of the ductus deferens and after survival times of 3, 4 and 5 days the spinal cord and brain were processed immunocytochemically. Virus-labelled neurones could be detected in the preganglionic sympathetic neurones and the dorsal commissural nucleus (upper lumbar segments) and in the sacral parasympathetic nucleus (L6 S1). Virus-infected perikarya were present in several brain stem nuclei including the gigantocellular and paragigantocellular nucleus, the lateral reticular nucleus, the nucleus of the solitary tract, the caudal raphe nuclei, the A1/C1, A2, A5 and A7 noradrenergic cell groups and the locus coeruleus. In the hypothalamus significant numbers of virus-infected neurones could be detected in the paraventricular nucleus. In most cases moderate numbers of virus-labelled cells were present in the lateral hypothalamic area, in the retrochiasmatic area, in the periventricular region and in the median preoptic area. Double-labelling immunofluorescence detection of virus-infected neurones and thyrosine hydroxylase (TH) showed colocalization of virus protein and TH in portion of neurones of the A1/C1, A2, A5 and A7 noradrenergic cell groups, in the locus coeruleus and in the hypothalamic paraventricular nucleus. The present results provide the first morphological data on the multisynaptic circuit of neurones innervating the ductus deferens. PMID- 12641828 TI - Effects of NCX 4050, a new NO donor, in rabbit and human corpus cavernosum. AB - The effects of NCX 4050, a drug belonging to a new class of NO donors, was investigated in isolated preparations of human and rabbit corpus cavernosum (CC) and in human foetal corpora cavernosa (hfCC) smooth muscle cells. In strips of rabbit CC, NCX 4050 (0.001-100 microM) induced a concentration-dependent relaxation which was influenced neither by Nw-nitro-l-arginine-methyl-ester (l NAME; 100 microm) nor by endothelium deprivation. The NCX 4050-induced relaxation was significantly reduced by the guanylate cyclase inhibitor 1H-[1,2,4] oxadiazolo[4,3-a]quinoxalin-1-one (ODQ; 1 microm) and enhanced by a specific phosphodiesterase 5 inhibitor, sildenafil (300 nm). Moreover, NCX 4050 (0.01-1 microm), induced a concentration-dependent potentiation of the relaxant response induced by electrical field stimulation (EFS) in rabbit preparations pre-treated with guanethidine and indomethacin. The relaxant effect of NCX 4050 was similar to that obtained by increasing concentrations (0.001-100 microm) of sodium nitroprusside (SNP) in either rabbit or human preparations. To further investigate the activity of NCX 4050 on human corpora cavernosa, we exposed cultured hfCC smooth muscle cells to increasing concentrations of NCX 4050 and SNP. We found that both compounds dose-dependently reduced cell proliferation. The antiproliferative effect of all the concentration tested of NCX 4050 was completely blocked by ODQ (1 microm). These results suggest that in rabbit and human corpora cavernosa NCX 4050 acts by activating guanylate cyclase activity, induces smooth muscle relaxation and quiescence. Our results provide a rationale for a possible future use of NCX 4050 in the pharmacotherapy of erectile dysfunction linked to an impaired release of NO from the endothelium. PMID- 12641830 TI - Seminal immunoreactive relaxin in domestic animals and its relationship to sperm motility as a possible index for predicting the fertilizing ability of sires. AB - Although immunoassayable relaxin has been detected in human and boar seminal plasma, there is no evidence suggesting the existence of immunoreactive relaxin in the seminal plasma of other domestic animals. The first objective of this study was to determine whether immunoreactive relaxin was present in the seminal plasma of bulls, rams and he-goats. In addition, the correlation of immunoreactive relaxin with sperm motility as an index for predicting the fertilizing ability of bull sires was investigated. Semen with normal sperm motility was collected from bulls, rams and he-goats, and the relaxin immunoreactivity of the semen samples was measured using a time-resolved fluoroimmunoassay (TR-FIA) for porcine relaxin that we developed. The presence of relaxin immunoreactivity was demonstrated in seminal plasma from bulls, rams and he-goats. The level of immunoreactive relaxin in seminal plasma was the highest in bulls followed by humans, rams, boars and he-goats in that order, when relaxin levels in boar and human semen having normal sperm motility were also assayed under the same conditions. When the correlation between the seminal plasma level of immunoreactive relaxin and sperm motility was examined in bull semen samples as an index for predicting fertilizing ability, it was found that the relaxin level was significantly correlated with the percentage of spermatozoa showing the most intensive motility (r = 0.64, p < 0.05). These results indicate that immunoreactive relaxin is widely found in the seminal plasma of domestic animals and that measuring the relaxin concentration of seminal plasma may be useful to identify subfertile sires or predict the fertility potential of individual sires. PMID- 12641829 TI - Participation of superoxide anion in the capacitation of cryopreserved bovine sperm. AB - Mammalian spermatozoa must undergo a preparation period known as capacitation to become capable of fertilizing oocytes. Controlled amounts of reactive oxygen species (ROS), such as superoxide anion (O2.-) and hydrogen peroxide (H2O2) have been shown essential for capacitation and acrosome reaction. The presence of an oxidase in the sperm plasma membrane has been suggested. The objective of the present study was to provide evidence for the production of O2.- by capacitating cryopreserved bovine spermatozoa. Percentages of capacitation and acrosome reaction were determined by the chlortetracycline assay. The effect of several nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitors on capacitation was also studied. H2O2 production was determined by the fluorometric assay using the p-hydroxyphenylacetic acid-horseradish peroxidase system. Superoxide dismutase (SOD) activity was determined spectrophotometrically at 480 nm. Heparin-dependent capacitation was inhibited by all NADPH oxidase inhibitors tested (p < 0.05). Significant levels of H2O2 were produced during capacitation with heparin; such production was inhibited by diphenyleneiodonium, one of the NADPH oxidase inhibitors. The addition of catalase to the incubation medium failed to modify the capacitation rate; inhibition was only observed when SOD was present (p < 0.05). Endogenous SOD activity was diminished during heparin dependent capacitation (p < 0.05). Similar levels of acrosome reaction induced by lysophosphatidylcholine were obtained in both heparin and O2.--dependent capacitation. Overall results suggest the participation of a sperm oxidase in bovine sperm capacitation. H2O2, generated by O2.- dismutation, failed to participate in capacitation, although this ROS may have been able to decrease endogenous SOD activity. Exogenous O2.- promotes physiological capacitation in cryopreserved bovine sperm, thus allowing the acquisition of fertilizing capacity. PMID- 12641831 TI - The role of the c-kit receptor in the regenerative differentiation of rat Leydig cells. AB - To determine the physiological role of the c-kit receptor, which is highly expressed in Leydig cells, the regenerative differentiation of Leydig cells was studied following transient degeneration induced by ethane dimethyl sulphonate (EDS) in c-kit-deficient mutant rats (Ws/Ws). EDS caused the destruction of Leydig cells; their functional recovery was evaluated by the weight change of the target organs of androgens, which occurred at the same rate in Ws/Ws and wild type rats. These results indicate that the tyrosine kinase activity of the c-kit receptor does not play an essential role in the regenerative differentiation of Leydig cells. PMID- 12641832 TI - Reactive oxygen species generation by human spermatozoa. PMID- 12641834 TI - Development of the angle of the anterior chamber in human and vertebrate animals' eyes. PMID- 12641835 TI - Cataracts in the Bichon Frise. AB - PURPOSE: To determine the clinical characteristics of possible inherited cataract in the Bichon Frise breed. These characteristics include the relative frequency, gender effects, site of first cataract formation, age of onset, relationship of age to cataract maturity, and other concurrent pre- and postoperative ophthalmic diseases. METHODS: Four different populations of Bichon Frise were examined. They included: (1) referred patients of the University of Florida (VMTH; 1990-2000); (2) patients from other universities and large institutions (VMDB; 1970-2000); (3) patients from the Canine Eye Registry Foundation (CERF; 1970-2000); and (4) special patients recruited from eye clinics and ACVO specialty practices (1995 2001). Blood samples were obtained from many patients for future DNA analyzes. Statistical comparisons between groups were by general linear and anova analyzes, and P < 0.05 was considered significant. RESULTS: The four populations of cataractous and total Bichon Frise dogs included: (1) UF-VMTH: 57 cataractous dogs; (2) VMDB: 406 dogs (28%) with cataracts; total dogs--1407; (3) CERF: 505 cataractous dogs (6%); total dogs--8222; and (4) ACVO: 223 cataractous dogs (57%); total dogs--391. In each population group, gender did not affect cataract distribution. Dogs between 2 and 8 years of age were most frequently affected, and initial cataract involvement affected equally the anterior and posterior cortices. Immature cataracts occurred more often in younger dogs, and hypermature cataracts were more frequently diagnosed in older dogs. Dogs with early cataracts were encountered more frequently in the CERF population. Pre- and postoperative retinal detachments (RD) were not infrequently diagnosed in the UF-VMTH and VMDB groups. RD in the UF-VMTH and VMDB groups occurred in 33% and 13% of the patients, respectively. CONCLUSIONS: Cataracts were first encountered in CERF and VMDB data between 1975 and 1979 and have increased since this time. As expected, both similarities and differences between the four different populations of Bichon Frise were encountered. Both sexes of dogs are equally affected. The anterior and posterior cortical areas of the lens were first involved and dogs 2 8 years of age are most frequently affected. Cataracts affected younger dogs in the CERF group and older cataractous dogs in the other populations. Cataract formation appears to be inherited in the Bichon Frise dog. The frequency of pre- and postoperative retinal detachments present higher risks for cataract surgery in this breed. PMID- 12641837 TI - Ultrasound biomicroscopy of the feline anterior segment. AB - Ultrasound biomicroscopy was performed on enucleated feline globes as a model for use in vivo. Quantitative measurements were obtained from the anterior segment using a 50-MHz transducer. Mean values from 26 feline globes were axial corneal thickness, 0.74 mm; anterior chamber depth, 4.20 mm; distance from the first ciliary process to the limbus, 3.17 mm; angle recess, 0.38 mm; angle opening distance, 1.05 mm; iris base width 0.38 mm; iris middle width 0.52 mm; iris tip width 0.13 mm, and iris-lens overlap, 0.69 mm. Distinctive features of the feline anterior segment include a relatively wide iridocorneal angle and deep anterior chamber. PMID- 12641836 TI - Symblepharon with aberrant protrusion of the nictitating membrane in the snowy owl (Nyctea scandiaca). AB - Two young snowy owl chicks were presented with aberrant protrusion of the nictitating membranes. This was caused by conjunctival adhesions causing symblepharon secondary to a previous septicemia episode. While symblepharon has been noted in birds before, this unusual presentation of the nictitating membrane has not been reported. Surgical intervention ameliorated the clinical signs, allowing vision in one bird by removal of the nictitating membranes, a technique which appeared to have no deleterious effects on the ocular surface. PMID- 12641838 TI - Early postnatal development of central corneal thickness in dogs. AB - OBJECTIVE: To investigate the changes in corneal thickness that occur during maturation of the canine eye over the first months of life. ANIMALS STUDIED: Dogs of two different breeds with ages ranging from 14 days to 42 weeks of age. PROCEDURES: The central corneal thickness was measured by ultrasonic pachymetry every week for the first month after eyelid opening (around 14 days) and then every month until 42 weeks of age. Segmented regression was applied to capture the two phases observed in the central corneal thickness plotted against age. Breed, eye and gender were also included in the model. RESULTS: Mean central corneal thickness (CCT) values initially decreased following eyelid opening, with the lowest point being reached at around 6 weeks of age. Then CCT gradually increased as the dogs matured. Differences between left and right eye were not significant. Breed and gender effects were significant factors in the statistical model. CONCLUSIONS: Following eyelid opening there is an initial decrease in corneal thickness until approximately 6 weeks of age, which presumably mirrors maturation of corneal endothelial cell function. After 6 weeks of age the CCT increases with age until approximately 30 weeks of age after which there was only a gradual increase over the remainder of the study period. A similar pattern of changes in corneal thickness in humans has been previously recorded. PMID- 12641839 TI - The effects of topical ocular application of 0.25% demecarium bromide on serum acetylcholinesterase levels in normal dogs. AB - OBJECTIVE: To assess the effects of topical ocular application of 0.25% demecarium bromide on serum acetylcholinesterase (AChE) levels in normal dogs. ANIMALS: Nine adult mixed breed dogs weighing between 18 and 27 kg. PROCEDURES: Fifty micro L of 0.25% demecarium bromide were applied to one eye of each dog every 8 h for 6 days. Blood was analyzed for AChE levels prior to commencement of eye drops, and at 45 min, 1 h 45 min, 4 h 45 min, 1 day, 3 days, and 7 days following commencement of eye drops using a 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) reaction. RESULTS: Acetylcholinesterase levels declined over the first 24 h following commencement of demecarium administration in most dogs. This decline was highly variable and was statistically significant by 24 h. In some individuals AChE levels were suppressed to levels approaching clinical toxicity. By day 3 AChE levels had risen to levels above baseline in most dogs. CONCLUSIONS: Topical ocular application of demecarium causes transient suppression of systemic acetylcholinesterase levels in most dogs. Acetylcholinesterase levels generally do not fall to toxic levels, but may do so in certain individuals. Demecarium bromide eye drops generally do not cause AChE toxicity, but dogs receiving such therapy should be monitored for signs of AChE toxicity, and concomitant use of other AChE inhibitors should be avoided. PMID- 12641841 TI - Structure and presumptive function of the iridocorneal angle of the West Indian manatee (Trichechus manatus), short-finned pilot whale (Globicephala macrorhynchus), hippopotamus (Hippopotamus amphibius), and African elephant (Loxodonta africana). AB - The iridocorneal angles of prepared eyes from the West Indian manatee, short finned pilot whale, hippopotamus and African elephant were examined and compared using light microscopy. The manatee and pilot whale demonstrated capacity for a large amount of aqueous outflow, probably as part of a system compensating for lack of ciliary musculature, and possibly also related to environmental changes associated with life at varying depths. The elephant angle displayed many characteristics of large herbivores, but was found to have relatively low capacity for aqueous outflow via both primary and secondary routes. The hippopotamus shared characteristics with both land- and water-dwelling mammals; uveoscleral aqueous outflow may be substantial as in the marine mammals, but the angular aqueous plexus was less extensive and a robust pectinate ligament was present. The angles varied greatly in size and composition among the four species, and most structures were found to be uniquely suited to the habitat of each animal. PMID- 12641840 TI - Flash electroretinography in standing horses using the DTL microfiber electrode. AB - PURPOSE: The goal of our study was the evaluation of a practical method for the recording of flash electroretinograms (ERGs) in sedated, standing horses with the DTL microfiber electrode. METHODS: The horses were sedated intravenously with detomidine hydrochloride (0.015 mg/kg). The pupil was dilated and the auriculopalpebral nerve was blocked. The ERGs were recorded with the active electrode on the cornea (DTL), the reference electrode near the lateral canthus, and the ground electrode over the occipital bone. The light intensities of the white strobe light were 0.03 cd x s/m2 (scotopic) and 3 cd x s/m2 (scotopic and photopic). Photopic and scotopic single flash and flicker responses to Ganzfeld stimulation were recorded. During the 20-min dark adaptation period the retina was stimulated every 5 min with the 0.03 cd x s/m2 single flash. RESULTS: The median b-wave amplitudes and implicit times were 38 microV and 33 ms (photopic cone-dominated response), 43 microV and 63 ms (5-min dark adaptation), 72 microV and 89 ms (10 min), 147 microV and 103 ms (15 min), 188 microV and 109 ms (20 min, 0.03 cd x s/m2, rod response), and 186 microV and 77 ms (20 min, 3 cd x s/m2, maximal combined rod-cone response). A steady increase in amplitude and implicit time was noted during dark adaptation. No oscillatory potentials could be isolated. CONCLUSIONS: The use of detomidine hydrochloride sedation and the DTL microfiber electrode allowed the recording of good quality ERGs. This protocol should permit the detection of functional problems in the retina without the risk involved with general anesthesia. PMID- 12641842 TI - Seasonal effects on the aerobic bacterial and fungal conjunctival flora of normal thoroughbred brood mares in Florida. AB - OBJECTIVE: To evaluate seasonal effects on the presence or absence of fungal and aerobic bacterial flora of the conjunctival fornix of normal Florida Thoroughbred horses. SAMPLE POPULATION: Both eyes of 100 horses. PROCEDURE: Horses with normal anterior segment ophthalmic examinations from three farms in north central Florida were included. Each animal had the ventral conjunctival fornix of each eye swabbed with sterile culturettes. Samples were taken in October, January, April, and July (1999-2000). Aerobic and fungal cultures were plated. Bacterial cultures were reviewed at 24 and 48 h. Fungal cultures were reviewed weekly for 4 weeks. Logistic regression analysis with season as a factor and age of the horse as a covariate was performed. Statistical significance was set at P < 0.01. RESULTS: Horses ranged from 3 to 24 years of age, with a median age of 9 years. Twenty-four genera of bacteria and 35 genera of fungi were recovered. Corynebacterium sp., Staphylococcus sp., Bacillus sp. and Moraxella sp. were the bacteria most frequently isolated. Mold species, dematiaceous mold species, Chrysosporium sp., Cladosporium sp., and Aspergillus sp. were the most frequently recovered fungi. Season did not have a significant effect on the presence of microorganisms isolated for individual horses adjusted for age. Younger horses had an increased incidence of gram-negative rods and fungal isolates. The number of bacteria and fungi isolated are not uniform across seasons. CONCLUSION: There were no significant differences between the number or type of organisms cultured during the sampling seasons in normal Florida horses. A large range of normal bacterial and fungal flora were isolated from these horses. The number of bacteria and fungi isolated are not uniform across seasons. The likelihood of detecting an organism depends on the horses' age. PMID- 12641843 TI - Fungal flora of normal eyes of healthy horses from the State of Rio de Janeiro, Brazil. AB - The conjunctival fungal flora of 32 adult horses with normal eyes (n = 64) from the State of Rio de Janeiro in Brazil was identified in the fall of 2000 using horses of different breeds, both genders and aged 5-19 years old. The culture samples were taken from the conjunctival sac of both eyes with a sterile cotton swab wetted with saline solution, seeded in Sabouraud's dextrose agar with chloramphenicol, and incubated for 5 days at an average temperature of 25 degrees C. The number of fungal colonies per eye varied between 0 and 250 colony forming units (CFUs). There were often differences in colony types between eyes of the same animal. Filamentous fungi of genera were isolated and identified in the following proportion of the total genera of fungal colonies isolated: Aspergillus (32.2%), Penicillium (25.8%), Scopulariopsis (15.9%), Trichoderma (11.2%), Cladosporium (5.6%), Mucor (2.1%), Syncephalastrum (2.1%), Eurotium (1.7%), Geotrichum (0.9%), Rhizopus (0.9%), Gliomastix (0.4%), Fusarium (0.4%), Staphylotrichum (0.4%) and Verticillium (0.4%). Yeast genera represented 9% of the total isolates. Over half the horses had at least one normal eye with either Aspergillus, Penicillium, Trichoderma or Scopulariopsis isolated, which is a departure from other studies of the normal horse eye. PMID- 12641844 TI - Neuroepithelial tumor of the retina in a dog. AB - ANIMAL STUDIED: At routine vaccination a tumor in the right eye of a Golden Retriever was diagnosed. PROCEDURE: The eye was enucleated, fixed in formalin and processed according to routine paraffin technique. Standard histologic, histochemical and immunohistochemical stainings were applied. Formalin-fixed tissue was used for electron microscopic examination. RESULTS: The tumor originated from the retina and was composed of two different morphologic parts, a retinoblastoma-like part of small neuroepithelial cells and a medulloepithelioma like part of columnar cells arranged in cords. DISCUSSION AND CONCLUSION: It is discussed whether the tumor had its origin in different clones or in one clone, and it is suggested that the double differentiation might be the result of a transformation of retinal cells, demasking stem cell-like potentials. PMID- 12641845 TI - Lens formation in the absence of optic cup in rat embryos irradiated with soft X ray. AB - In order to investigate the effect of soft X-ray irradiation on ocular development, pregnant rats were exposed to a single 12.5 Gy irradiation on embryonic day 9 (ED 9). The embryos obtained by laparotomy on ED 12 and 21 were examined for ocular abnormalities under a binocular stereo-microscope and a light microscope. The ED 12 embryos were stained with osmium tetroxide to facilitate the observation. The stereo-microscopic examination on ED 12 and 21 revealed various types of ocular abnormalities characterized primarily by aplasia or hypoplasia of the optic cup and invaginated lens placode. The light microscopic examination further confirmed these findings histomorphologically, and the hypoplastic abnormalities were classified into three types: (1) hypoplasia of the optic cup and invaginated lens placode, (2) complete malformation of the optic cup and hypoplasia of the invaginated lens placode, and (3) complete malformation of the optic cup and invaginated lens placode. Because the lens was formed in the complete absence of the retina, the development and differentiation of the retina and lens do not seem to be tightly synchronized. Thus, this sequential analysis on ocular abnormalities during the early stage of development supports the notion that the presence of the retina is not always necessary for the development of the lens. PMID- 12641846 TI - Inhibition of collagenase breakdown of equine corneas by tetanus antitoxin, equine serum and acetylcysteine. AB - OBJECTIVE: To determine whether tetanus antitoxin, equine serum, and acetylcysteine, which are currently used in the treatment of equine corneal ulcer, inhibit the digestion of equine corneal collagen when exposed to collagenase in vitro. ANIMALS STUDIED: Corneas from 40 adult horses. PROCEDURES: Sections of equine corneas were incubated with saline, a solution of bacterial collagenase in saline, bacterial collagenase in saline plus equine tetanus antitoxin, bacterial collagenase in saline plus equine serum, or bacterial collagenase in saline plus acetylcysteine. Each one of the collagenase inhibitors was tested at different concentrations. The degree of corneal collagen digestion was determined by concentrations of hydroxyproline released into the incubation media and/or by weight loss of the cornea. RESULTS: Corneas exposed to collagenase released a significant (0.05 level) large amount of hydroxyproline (43.1 +/- 2.3 microg/mL/100 mg cornea/5 h) and decreased cornea weight by up to 89%. Blood serum (200 microL/mL), purified albumin or globulin fractions of serum, tetanus antitoxin (120 units/mL), and acetylcysteine (20 mg/mL) when used at the highest concentrations blocked collagenase digestive activity by approximately 50%. Dilution of inhibitors decreased corneal protection and linearly increased corneal weight loss. Purified equine serum albumin and globulin fractions were equally effective in protecting corneas. CONCLUSIONS: This experiment indicates that tetanus antitoxin, serum and acetylcysteine equally protected corneas from collagenase digestion, in vitro. However, a clinical trial is needed to establish relative therapeutic value. PMID- 12641847 TI - Changes in intraocular pressure and pupil size following intramuscular administration of hydromorphone hydrochloride and acepromazine in clinically normal dogs. AB - OBJECTIVE: To investigate the effects of intramuscularly administered hydromorphone hydrochloride and acepromazine on intraocular pressure (IOP) and pupil size (PS). ANIMALS STUDIED: Seventeen dogs free of clinically relevant ocular abnormalities. PROCEDURE: Measurements of IOP and PS were obtained and the dogs were injected intramuscularly with hydromorphone (0.04-0.08 mg/kg) and acepromazine (0.04 mg/kg). Measurements of IOP and PS were repeated 10 min and 25 min later. RESULTS: Though a decreasing trend in IOP values was demonstrated, no significant difference was noted in IOP from the initial examination to examination following intramuscular administration of hydromorphone and acepromazine. Significant miosis was present in 16 of 17 dogs at 10 min and 25 min following administration of hydromorphone and acepromazine. CONCLUSION: Hydromorphone (0.04-0.08 mg/kg) and acepromazine (0.04 mg/kg) cause significant miosis in dogs at 10 and 25 min following intramuscular administration. PMID- 12641848 TI - Progressive changes in ophthalmic blood velocities in Beagles with primary open angle glaucoma. AB - OBJECTIVE: To measure changes in the ocular and orbital blood flow velocities by color Doppler imaging (CDI) in beagles with primary open angle glaucoma as the disease progressed from early to advanced stages. METHODS: CDI measurements were performed periodically on 13 glaucomatous Beagles during the nontreated mild, moderate and advanced stages of POAG over the course of 4 years. CDI was performed with the dogs lightly anesthetized (butorphanol 0.1 mg/kg IV, acepromazine maleate 0.02 mg/kg IV, and atropine sulfate 0.05 mg/kg) while the CD transducer was placed directly on the cornea anesthetized with 0.5% tetracaine hydrochloride. Intraocular pressure (IOP) by pneumatonography or TonoPen XL, heart rate and mean arterial blood pressure were measured at the beginning, middle and end of each study. The ophthalmic vessels examined included: external ophthalmic arteries and veins, long and short posterior ciliary arteries, anterior ciliary arteries and veins, primary retinal arteries, and vortex veins. Recordings of each vessel included peak systolic velocity (PSV), end diastolic velocity (EDV) and time averaged velocity (TAV), and when possible the resistive index (RI) and pulsatility index (PI) were computed. RESULTS: CDI abnormalities were present before intraocular pressure exceeded the normal range. As the animals aged, and the glaucoma progressed with higher levels of IOP, significant changes occurred in nearly all vessels, and generally included a major increase in RI (P < 0.001) and an increase in the PI (P < 0.001). Mean arterial blood pressure (105 +/- 18 mmHg) and heart rate (118 +/- 33/min) remained reasonably constant. The IOP gradually increased as the disease progressed (early and normotensive: 19.4 +/- 3.9 mmHg; moderate: 29.7 +/- 2 mmHg; and advanced: 44.5 +/ 6 mmHg). The ocular veins seemed most influenced early on in the disease. Late in the disease, ocular venous blood flow could not be consistently demonstrated. An increase in the PI of ocular veins occurred in the moderately and severely affected glaucomatous Beagles. As the IOP increased, there were trends of increasing resistive index and pulsatility index in most arteries, and periods of marked decreased velocities of the vortex and external ophthalmic veins in severe cases. CONCLUSION: CDI measurements in Beagles with primary open angle glaucoma during the course of 4 years indicate easily measurable and repeatable progressive blood flow abnormalities before the elevation of IOP and, thereafter, with gradually increased levels of IOP. PMID- 12641849 TI - Primary ocular osteosarcoma in a dog. AB - An adult male Rottweiler presented to the veterinary medical teaching hospital at Purdue University with a 1-month history of hyphema. On physical examination conjunctivitis, episcleral hyperemia, corneal edema, hyphema, mild glaucoma and loss of vision were observed in the left eye. No other abnormalities were found. The left globe was surgically removed because of the high likelihood of neoplasia and it was fixed in 10% buffered formalin and submitted for pathology. A histologic diagnosis of primary osteosarcoma of the eye was made. Radiographic evaluation did not reveal any evidence of other tumors or pulmonary metastasis. This is the fourth canine case of primary intraocular osteosarcoma to be documented. PMID- 12641856 TI - Additional considerations in longitudinal database research. PMID- 12641857 TI - The use of claims databases in pharmacoeconomic studies. PMID- 12641858 TI - A checklist for retrospective database studies--report of the ISPOR Task Force on Retrospective Databases. AB - INTRODUCTION: Health-related retrospective databases, in particular claims databases, continue to be an important data source for outcomes research. However, retrospective databases pose a series of methodological challenges, some of which are unique to this data source. METHODS: In an effort to assist decision makers in evaluating the quality of published studies that use health-related retrospective databases, a checklist was developed that focuses on issues that are unique to database studies or are particularly problematic in database research. This checklist was developed primarily for the commonly used medical claims or encounter-based databases but could potentially be used to assess retrospective studies that employ other types of databases, such as disease registries and national survey data. RESULTS: Written in the form of 27 questions, the checklist can be used to guide decision makers as they consider the database, the study methodology, and the study conclusions. Checklist questions cover a wide range of issues, including relevance, reliability and validity, data linkages, eligibility determination, research design, treatment effects, sample selection, censoring, variable definitions, resource valuation, statistical analysis, generalizability, and data interpretation. CONCLUSIONS: For many of the questions, key references are provided as a resource for those who want to further examine a particular issue. PMID- 12641859 TI - Indirect costs of respiratory syncytial virus hospitalizations: a commentary. PMID- 12641860 TI - Time and out-of-pocket costs associated with respiratory syncytial virus hospitalization of infants. AB - OBJECTIVE: The objective of this study was to quantify time spent plus out-of pocket costs associated with confirmed respiratory syncytial virus (RSV) hospitalization of infants not prophylaxed against RSV. METHODS: A prospective survey was carried out at multiple tertiary care hospitals in the United States. PATIENTS: The patients consisted of a consecutive sample of infants <12 months, born between 33 and 35 weeks of gestation. One site also enrolled full-term infants hospitalized with confirmed RSV. Daily patient census identified eligible patients. Consenting caregivers of eligible subjects (n=84, 1 refusal) were interviewed on discharge day and by telephone approximately 30 days following discharge regarding time and out-of-pocket costs due to RSV. RESULTS: Total average out of pocket expenses were 643.69 US dollars (range 21-16,867 US dollars; SD 2,403 US dollars) for premature and 214.42 US dollars (range 6-827 US dollars; SD 218 US dollars) (P=.0158) for full-term subjects. Total average economic burden per admission was 4517.07 US dollars for premature and 2135.30 US dollars for full-term infants, including the value of lost productivity but excluding inpatient hospital and physician bills and lost income. Premature infants (n=48) had longer hospital stays (mean 6.9 days; SD 7.5 vs. 3.4 days; SD 2.6 days) (P=.001) with an associated mean total time spent by up to 5 adults of 281.7 hours (range 25-2819.7 hours; SD 465.8 hours) versus a mean of 139.7 hours (range 31.8-561.3 hours; SD 118.1 hours) for term infants (P=.109). Time and out of-pocket costs continued after discharge. CONCLUSIONS: RSV hospitalization of infants is associated with substantial, previously unmeasured time and monetary losses. These losses continued following discharge. The economic burden on families and society appears heavier for infants born at 33 to 35 weeks of gestation than for full-term infants. PMID- 12641861 TI - Economic costs of influenza-related work absenteeism. AB - BACKGROUND: Influenza vaccinations are currently advocated only for individuals over age 50. However, vaccination of all working-age people may be warranted based on reduced absenteeism from work. OBJECTIVE: This study aims to quantify the association between lost workdays and influenza, controlling for other factors. A secondary aim of the study is to assess the net benefit of expanded vaccination in a workplace setting. RESEARCH DESIGN: Multivariate regression analyses of the 1996 Medical Expenditure Panel Survey Household Component are used to estimate the number of workdays missed because of influenza-like illness (ILI) when controlling for other health, demographic, and employment factors. Mean productivity costs are measured in terms of absences from work and valued in dollar terms. The net benefit of influenza vaccination is estimated using a simple decision analysis. SUBJECTS AND MEASURES: Health, demographic, and employment data for employed individuals between the ages of 22 and 64 years are analyzed. RESULTS: The average number of workdays missed due to ILI was 1.30 days, and the average work loss was valued at 137 US dollars per person. The vaccine strategy was not preferred in the baseline analysis; however, this result was sensitive to assumptions regarding the incidence of influenza, the cost of delivering the vaccine, and the productivity impact of worker absenteeism. Moreover, nonproductivity benefits of vaccination were omitted. CONCLUSIONS: The economic attractiveness of expanded investment in influenza vaccination hinges on employer- and population-specific assumptions. Our analysis provides a simple framework within which competing considerations of disease epidemiology, worker productivity, and economic cost may be weighed. PMID- 12641862 TI - Oseltamivir for treatment of influenza in healthy adults: pooled trial evidence and cost-effectiveness model for Canada. AB - BACKGROUND: Influenza is a common viral respiratory infection that is associated with significant morbidity. Oseltamivir (Tamiflu) is a neuraminidase inhibitor-a new class of antiviral treatment for influenza where efficacy and safety has been established but cost-effectiveness is unknown. METHODS: A decision analytic model was used to estimate the costs and effectiveness of two treatment scenarios for empiric management of otherwise healthy nonelderly patients, presenting with influenza-like illness (ILI) to primary care physicians in Canada: 1) where oseltamivir is reimbursed and on formulary for prescription; and 2) where oseltamivir is not on formulary. Outcomes are influenza-days averted and quality adjusted life-years (QALYs) gained. Effectiveness, utility, and pneumonia complication risk estimates are by pooled analysis of patient-level data from four clinical trials. Unit cost information (Canadian dollars) was obtained from published sources in Ontario. Probabilistic sensitivity analysis was conducted using Monte Carlo simulation. RESULTS: Of 2288 patients randomized, influenza was confirmed in 1575 (69%) and oseltamivir treatment reduced the mean time to symptom alleviation by 1.08 days (95% confidence interval CI] 0.58-1.59). Infected patients treated with oseltamivir had higher utility scores (quality of life) than placebo patients over the 7 days of follow-up (P <.05). Cost per influenza-day averted with oseltamivir on formulary is 49 US dollars (95% CI 31 107) and the cost per QALY is 57,863 US dollars (95% CI 48,919- 70,149 US dollars). Results are sensitive to the percentage of patients presenting to their physician beyond 48 hours from symptom onset who get oseltamivir and the prevalence of influenza among patients presenting with ILI. CONCLUSIONS: Oseltamivir for treatment of patients with ILI is potentially cost-effective if clinical diagnostic specificity for influenza observed in clinical trials is applicable to routine practice. More population-based information on the prevalence of influenza among early (<48 hours) presenters with ILI would be valuable. PMID- 12641863 TI - Cost-effectiveness of enoxaparin as thromboprophylaxis in acutely ill medical patients in Spain. AB - OBJECTIVE: The objective of this study was to determine the cost-effectiveness of thromboprophylaxis with enoxaparin versus no thromboprophylaxis in patients with acute medical illness in Spain from the society perspective. METHODS: Markov process analysis techniques were used to model the health economic outcomes. Clinical data were derived mainly from the MEDENOX trial, while health-care utilization was derived from Delphi panels. RESULTS: An analysis over the MEDENOX trial period shows that the cost per event avoided is currency 432, while the cost per life saved is currency 1527. The cost per event includes all medical resource utilization costs associated with the event. The lifetime model, which assumes no higher risk for recurrence of venous thromboembolism (VTE) and mortality in asymptomatic patients, shows that the use of enoxaparin leads a cost per event avoided of currency 270 and cost per life-year gained of currency 71. If the lifetime model assumes a higher risk for recurrence of VTE in asymptomatic patients, enoxaparin is dominant over no thromboprophylaxis. CONCLUSION: The results showed that the favorable clinical benefit of enoxaparin as thromboprophylaxis in patients with acute medical illness, which was observed in the MEDENOX trial, results in a positive health economic benefit in both the short term and the long term in the health-care setting of Spain. PMID- 12641864 TI - Economic consequences of renal dysfunction among cardiopulmonary bypass surgery patients: a hospital-based perspective. AB - BACKGROUND: Renal dysfunction is common after cardiopulmonary bypass procedures and is associated with higher mortality rates and longer lengths of stay. However, less is known about the actual cost of care for these patients. We sought to quantify the hospital costs attributed to renal dysfunction in cardiopulmonary bypass patients at a large academic referral center. METHODS: All patients undergoing cardiopulmonary bypass procedures were identified through administrative databases for a 3-year study period. Renal failure was defined using laboratory values from the hospitalization. Total direct costs and costs by hospital department were determined using the hospital cost-accounting system. A multivariate linear model was developed to determine total direct hospital costs after cardiopulmonary bypass procedures after adjusting for relevant clinical and demographic variables. RESULTS: Nine percent of the study population developed new-onset renal dysfunction, and 18% of patients overall undergoing cardiopulmonary bypass experienced renal dysfunction during the hospitalization. Direct costs were 42% higher for patients with renal dysfunction, an average of 5807 US dollars per case. Higher costs were noted in intensive care unit use and pharmacy, laboratory, and radiology services. CONCLUSIONS: Renal dysfunction increases the direct hospital costs of care, even after adjustment for age, sex, race, and comorbidities. Nationwide, we estimate that renal dysfunction may add up to 643 million US dollars in hospital costs for caring for cardiopulmonary bypass patients. Interventions designed to reduce the incidence and severity of renal dysfunction may significantly reduce hospital costs for these patients. PMID- 12641865 TI - The cost-effectiveness of acetaminophen, NSAIDs, and selective COX-2 inhibitors in the treatment of symptomatic knee osteoarthritis. AB - OBJECTIVE: The objective of this study was to conduct an economic evaluation of rofecoxib and celecoxib compared with high-dose acetaminophen or ibuprofen with and without misoprostol for patients with symptomatic knee osteoarthritis (OA). METHODS: A decision analysis model was designed over 6 months using two measures of effectiveness: 1) number of upper gastrointestinal (GI) adverse events averted; and 2) number of patients who achieved perceptible pain relief. Separate analyses were conducted for all patients and for those who did not respond to acetaminophen. Outcome probabilities were obtained from a comprehensive review of randomized controlled trials and observational studies. Costs were derived from actual resource utilization of OA patients. RESULTS: In terms of averting GI events, acetaminophen dominates the other options for an average risk patient population. For patients who did not respond to acetaminophen, rofecoxib had the lowest incremental cost-effectiveness ratio (ICER) per GI event avoided (32,000 US dollars) relative to ibuprofen. In terms of pain control, ibuprofen had an ICER of 610.77 US dollars per additional patient achieving minimal perceptible clinical improvement (MPCI) relative to acetaminophen, while rofecoxib had an ICER of 12,000 US dollars relative to ibuprofen. For patients who did not respond to acetaminophen and who are at high risk of developing an adverse GI event, rofecoxib dominates ibuprofen as the preferred alternative for both measures of effectiveness. One-way, two-way, and probabilistic sensitivity analyses established that these results were generally robust. CONCLUSIONS: Our results suggest that for average-risk knee OA patients, acetaminophen dominates the other therapies in terms of cost per GI event averted. In terms of pain relief, cost effectiveness acceptability curves indicate that if one values pain relief below 275 US dollars per patient achieving MPCI, acetaminophen is the therapy most likely to be optimal; between 275 US dollars and 14,150 US dollars, ibuprofen is most likely to be optimal; and above 14,150 US dollars, rofecoxib is most likely to be optimal. PMID- 12641866 TI - Economic comparison of home-care-based versus hospital-based treatment of chemotherapy-induced febrile neutropenia in children. AB - OBJECTIVE: The purpose of this study was to compare health-care resource utilization and outcomes among children treated for low-risk febrile neutropenia (FN) in a hospital-based setting with those treated in a home-care-based setting. METHODS: The perspective of this retrospective, cohort study was the health payer. We collected health-care utilization and treatment outcome data from medical records of 63 children (26 boys and 37 girls) with low-risk, chemotherapy induced FN who were treated at the University of Arizona (27 children, the hospital-based group) and University of New Mexico (36 children, the home-care based group). We identified 144 FN episodes (72 episodes in each group). Health care utilization included physician visits, home-care visits, laboratory visits, outpatient visits, hospital days, intensive care unit days, medical tests and studies, and medications used to manage FN (e.g., filgrastim, antimicrobials, and ancilliary drugs and supplies). We applied uniform charges, based on those used at the University of New Mexico in 1998. We collected outcomes of the FN treatment (success vs. failure and time to resolution, defined as number of days of antibiotic therapy). Rates of positive blood cultures during treatment were also compared. Data were analyzed using nonparametric Mann-Whitney U tests for continuous data and chi-square analysis for categorical data. Sensitivity analyses were conducted by varying the amount of total resource utilization, as well as utilization of specific health-care resources. RESULTS: There was no difference in outcome; all episodes of treatment in both groups resulted in successful recovery from FN. Time to resolution of FN was 8.3 +/- 2.7 days for home-care FN episodes versus 7.3 +/- 3.6 days for hospital FN episodes (P =.064). Median charge per FN episode was significantly (P<.001) greater when managed in the hospital compared to home care (9392 US dollars vs. 5893 US dollars). There was greater use of laboratory and radiographic studies in the hospital-based patients (P <.01). However, children in the home-care-based group were more often treated with granulocyte colony-stimulating factor (filgrastim, median charge 1085 US dollars vs. 451 US dollars, P <.001), and median antibiotic charges were higher (2523 US dollars vs. 1526 US dollars, P <.001). Positive blood cultures were more common among the hospital-based FN treatments (30.6 vs. 11.1%, P=.012). CONCLUSIONS: We found that management of low-risk FN in a home-care-based setting was associated with significantly lower median total charges with no differences in outcome. PMID- 12641868 TI - Rational formulary decision making: is drug classification rational? PMID- 12641867 TI - Costs of drug delivery for CHOP, COP/CVP, and fludarabine: an international assessment. AB - OBJECTIVES: The purpose of this analysis was to assess the real-life direct costs of drug delivery for frequently used chemotherapeutic regimens in patients with relapsed low-grade non-Hodgkin's lymphoma (NHL). METHODS: This was a retrospective analysis of direct costs of drug delivery (acquisition plus administration) of relapsed low-grade NHL in 424 patients in Canada, Germany, and Italy. Results were expressed as an average treatment cost per patient for six cycles of chemotherapy. Exchange rates used were $1 (Canada)= currency 0.672, 1 DM (Germany)= currency 0.511, and 1 Lit (Italy)= currency 0.000517. RESULTS: Direct costs of drug delivery were greater for inpatients receiving fludarabine (Canada currency 12,669; Italy currency 13,027) than for CHOP (Canada currency 7856; Germany currency 7218; Italy currency 4251) or COP/CVP (Canada currency 7360; Germany currency 8449). Treatment administration setting was a major cost driver with inpatient treatment up to 9-fold more expensive than the same regimen given to outpatients. Drug administration costs comprised the largest proportion of the total for each regimen in the inpatient setting (69-98%). Costs of drug delivery in the outpatient setting were 10% to 65% of those in the inpatient setting. Again, fludarabine was more expensive (Italy currency 8493; Canada currency 7269) than CHOP (Canada currency 4403; Germany currency 2150; Italy currency 1264) and COP/CVP (Canada currency 3009; Germany currency 867). Administration costs were 2.5- to 15-fold higher for inpatients compared to outpatients. CONCLUSIONS: Costs of drug administration are a major driver for total direct treatment costs in the treatment of relapsed low-grade NHL and are at least as important as drug acquisition costs. Drug administration practices, in terms of inpatient or outpatient treatment, are a major factor in determining overall direct costs. Therapeutic strategies, which offer shortened treatment duration and/or a simple mode of administration, are likely to be economically attractive. PMID- 12641869 TI - The clinical epidemiology of cardiovascular disease in chronic kidney disease. PMID- 12641870 TI - Cardiovascular disease in chronic renal failure: pathophysiologic aspects. AB - Cardiovascular complications are the leading cause of mortality in patients with end-stage renal disease (ESRD). The excess cardiovascular risk and mortality is already demonstrable in early renal disease and in patients with chronic renal failure (CRF), with the highest relative risk of mortality in the youngest patients. The high risk for cardiovascular disease (CVD) results from the additive effect of multiple factors, including hemodynamic overload and several metabolic and endocrine abnormalities more or less specific to uremia. CVD includes disorders of the heart (left ventricular hypertrophy [LVH], cardiomyopathy) and disorders of the vascular system (atherosclerosis, arteriosclerosis), these two disorders being usually associated and interrelated. LVH is the most frequent cardiac alteration in ESRD, resulting from a combined pressure and volume overload. LVH in general is an ominous prognostic sign and an independent risk factor for arrhythmias, sudden death, heart failure, and myocardial ischemia. Regression of LVH needs a combined intervention to reduce hemodynamic overload and is associated with improved prognosis and survival. Clinical studies have shown that damage of large conduit arteries is a major contributing factor for the high incidence of congestive heart failure (CHF), LVH, ischemic heart disease (IHD), sudden death, cerebrovascular accidents, and peripheral artery diseases. Damage to large conduit arteries is principally related to highly calcified occlusive atherosclerotic lesions and to stiffening of large capacitive arteries. These two complications are independent risk factors for survival, and improvement of arterial stiffness is associated with better prognosis and survival. The present review summarizes the most recent works dealing with the pathophysiology of CVD and some aspects of the therapeutic approach. PMID- 12641871 TI - Is chronic kidney disease a cardiovascular disease risk factor? AB - Cardiovascular disease (CVD) is prevalent in patients with chronic kidney disease (CKD) and may account for 50% of all deaths. The recent Dialysis Outcomes Quality Initiative (DOQI) publication on the evaluation, classification, and stratification of CKD states that a reduced glomerular filtration rate (GFR) identifies individuals at greater risk for CVD and death. This risk is the result of traditional and nontraditional CVD risk factors. However, the relative contribution of these risk factors in the CKD population remains uncertain. Recently interest in kidney disease (reduced GFR) as an independent nontraditional risk factor for CVD has come to the forefront. Studies examining this potential link have included community-based cohort studies, studies in patients with extensive comorbidity, and reports in kidney transplant recipients. Herein, results from these studies are reviewed. The difference between an independent CVD risk factor and a causal CVD risk factor is discussed, with particular emphasis on the temporal association between exposure (GFR) and outcome (CVD and CVD death). PMID- 12641872 TI - Clinical epidemiology of cardiovascular disease in chronic kidney disease prior to dialysis. AB - Cardiovascular disease (CVD) remains the leading cause of morbidity and mortality in patients with end-stage renal disease (ESRD). Both in dialysis and in transplant patients, CVD remains the leading cause of death. There is accumulating evidence that the increase in CVD burden is present in patients prior to dialysis, due to both conventional risk factors as well as those specific to kidney disease. Of importance is that even in patients with mild kidney disease, the risk of cardiovascular events and death is increased relative to patients without evidence of kidney disease. The new classification system proposed by the National Kidney Foundation as part of the Dialysis Outcomes Quality Initiative (DOQI) process describes the five stages of kidney disease, as well as those complications associated with chronic kidney disease (CKD), in particular cardiovascular risk factors and disease. Patients with kidney disease are deemed to be at highest cardiovascular risk. CVD, defined as the presence of either congestive heart failure (CHF), ischemic heart disease (IHD), or left ventricular hypertrophy (LVH), is prevalent in cohorts with established CKD (8 40%). The prevalence of hypertension, a major risk factor for coronary artery disease (CAD) and LVH is high in patients with CKD (87-90%). At least 35% of patients with CKD have evidence of an ischemic event (myocardial infarction or angina) at the time of presentation to a nephrologist. The prevalence of LVH increases at each stage of CKD, reaching 75% at the time of dialysis initiation, and the modifiable risk factors for LVH include anemia and systolic blood pressure, which are also worse at each stage of kidney disease. Even under the care of nephrologists, a change in cardiac status (worsening of heart failure or anginal symptoms) occurs in 20% of patients. The presence of CVD predicts a faster decline of kidney function and the need for dialysis, after controlling for all other factors including glomerular filtration rate (GFR), age, and the presence of LVH. This article describes the new classification system for staging of CKD, defines and describes CVD in CKD, and reviews the evidence and its limitations with respect to the current understanding of CKD and CVD. Specifically, methodologic issues related to survival and referral bias limit our current understanding of the complex interaction of conventional and nonconventional kidney disease-specific risk factors. We identify the importance of well-conducted studies of patient groups with and without CVD, with and without CKD, in order to better understand the complex physiology so that treatment strategies can be appropriately applied. PMID- 12641873 TI - Clinical epidemiology of cardiac disease in renal transplant recipients. AB - Cardiovascular disease (CVD) is the major cause of death among renal transplant recipients (RTRs), accounting for 17-50% of deaths. Both cardiomyopathy (congestive heart failure [CHF] and left ventricular hypertrophy [LVH]) and ischemic heart disease (IHD) are important complications of renal transplantation, although the morbid impact of cardiomyopathy has been overlooked until recently. Echocardiographic disorders and clinical CHF occur far more frequently in RTRs than in the general population, suggesting that renal transplantation may be a state of accelerated heart failure. In contrast, the incidence of IHD in RTRs is similar to that in the Framingham cohort. Age, diabetes, and gender remain important markers of risk for both disorders. Smoking, hyperlipidemia, and hypertension appear to be the major reversible risk factors for IHD, while anemia and hypertension are major reversible risk factors for cardiomyopathy. Definitive evidence on optimal intervention is lacking. Clinical trials are needed to define optimum targets for treatment of these risk factors, especially hypertension and anemia. PMID- 12641874 TI - Clinical epidemiology of cardiac disease in dialysis patients: left ventricular hypertrophy, ischemic heart disease, and cardiac failure. AB - Patients with end-stage renal disease (ESRD) are at extreme cardiovascular risk. At least half of all patients starting dialysis therapy have overt cardiovascular disease (CVD). In addition, recent studies suggest annual incidence rates for new onset cardiac failure, peripheral vascular disease, ischemic heart disease (IHD), and stroke of approximately 7%, 7%, 5%, and 1%, respectively. High-level exposure to traditional risk factors, such as smoking and dyslipidemia, hemodynamic overload factors, such as anemia and hypertension, and a myriad of metabolic factors related to uremia are all likely to play a role. There has been explosive growth in observational studies and a heartening, if less dramatic, increase in risk factor intervention trials, suggesting that risk factor modification can lead to meaningful benefit. PMID- 12641876 TI - Traditional cardiac risk factors in individuals with chronic kidney disease. AB - Individuals with chronic kidney disease (CKD) are at increased risk for the development and progression of cardiovascular disease (CVD). The increased risk is due to a higher prevalence of both traditional risk factors as well as nontraditional risk factors. In this review we focus on individuals at all stages of CKD and discuss modifiable traditional risk factors, namely hypertension, dyslipidemia, diabetes mellitus and poor glycemic control, smoking, and physical inactivity. The prevalence of each risk factor and its relationship with CVD is described. Treatment recommendations are provided using evidence available from populations with CKD or evidence extrapolated from the general population when there are insufficient data on individuals with CKD. PMID- 12641877 TI - Hemodynamic cardiovascular risk factors in chronic kidney disease: what are the effects of intervention? AB - Left ventricular (LV) volume and pressure overload occur frequently in chronic kidney disease (CKD). Anemia is a risk factor for left ventricular hypertrophy (LVH) and dilatation, heart failure, and death. Normalization of hemoglobin with erythropoietin may prevent LVH and dilatation in CKD, but in patients in later phases of their cardiac disease, this intervention is not of benefit. Increased vascular volume causes hypertension, which in turn causes LVH, cardiac failure, and ischemic heart disease (IHD). Manifestations of arteriosclerosis are associated with adverse cardiac outcomes, and angiotensin converting enzyme (ACE) inhibitors may improve LVH and markers of arteriosclerosis. Aortic stenosis in dialysis patients occurs infrequently, but may deteriorate rapidly. The hemodialysis milieu is the quintessential model of LV overload cardiomyopathy. PMID- 12641878 TI - Calcium phosphate metabolism and cardiovascular disease in patients with chronic kidney disease. AB - Traditional risk factors for atherosclerotic cardiovascular disease (CVD) do not adequately explain the considerable increase in cardiovascular mortality observed among patients with end-stage renal disease (ESRD): these patients experience mortality rates 10-100 times those without ESRD. Disorders of mineral metabolism, including abnormalities in calcium, phosphorus, parathyroid hormone, and vitamin D, represent cardiovascular risk factors unique to the ESRD population. These disturbances manifest clinically through the promotion of extraskeletal calcification and disorders of bone remodeling, two processes which appear to share a common pathogenesis. This article presents evidence describing the impact of calcification-induced arterial stiffness on cardiovascular outcomes of patients with ESRD, along with data relating altered mineral metabolism to all cause and cardiovascular mortality. Specific management recommendations include 1) early intervention to prevent the development of overt secondary hyperparathyroidism, 2) a more judicious strategy for vitamin D therapy, and 3) a thoughtful approach to the use of calcium-containing phosphate binders, taking into account the underlying bone remodeling disorder and the presence or absence of extraskeletal calcium accumulation. PMID- 12641880 TI - Uremia-related metabolic cardiac risk factors in chronic kidney disease. AB - Growing evidence has been gathered over the last 15 years regarding the role of nontraditional or uremia-related risk factors in the pathogenesis of atherosclerosis in subjects with renal failure. Among those factors, dyslipidemia, inflammation, hyperhomocysteinemia, and oxidant stress have been extensively studied. However, the clinical significance of many of these factors remains controversial in light of reported studies. In this article, the existing evidence regarding the role of uremia-related risk factors in the pathogenesis of atherosclerosis is reviewed, with special emphasis on prevalence, cardiac risk, and management in patients with chronic kidney disease (CKD). Consensus treatment recommendations are provided for risk factors for which there is evidence to support preventive or therapeutic interventions. PMID- 12641881 TI - Applying multiple interventions in chronic kidney disease. AB - The medical, social, and financial burdens posed by end-stage renal disease (ESRD) are many and growing rapidly. People generally reach ESRD as a result of chronic progressive kidney disease. Advancing kidney disease is associated with several treatable complications, which if poorly managed reduce the length and quality of life. In addition, there are strong links between chronic kidney disease (CKD) and cardiovascular disease (CVD). Many people with less advanced CKD will die or suffer complications of CVD before reaching ESRD. Efficacious interventions, such as lowering blood pressure and treating dyslipidemia, can substantially reduce the progression of both kidney and cardiovascular disease. Careful management of these complex and interrelated diseases and risk factors requires detailed longitudinal and focused care which does not seem to be optimally delivered by health service practitioners organized in traditional ways. A disease management approach offers promise in this setting, but requires further study of clinical and economic impact. PMID- 12641882 TI - Management of heart failure and coronary artery disease in patients with chronic kidney disease. AB - Cardiovascular disease (CVD) is a major contributor to the mortality and morbidity of patients who suffer from chronic kidney disease (CKD). Heart failure and ischemic heart disease (IHD) are both highly prevalent in this population. The diagnosis of myocardial dysfunction is usually based on echocardiography. As in the general population, systolic dysfunction is treated with a combination of diuretics, renin-angiotensin system blockade, and beta-receptor antagonists. Diastolic dysfunction is best managed by eliminating the cause. Non-invasive tests for coronary artery disease (CAD) may be less reliable in patients with renal disease compared with nonuremic patients. Medical therapy of IHD in this population is generally similar to that for other patient groups, but surgical revascularization appears to carry a higher risk of complications with poorer clinical outcomes. The choice of revascularization procedure (coronary artery bypass grafting versus percutaneous transluminal angioplasty) should be based on the specific coronary anatomy of a given patient as well as a consideration of other comorbid factors. PMID- 12641883 TI - Guidelines for training, certification, and accreditation for hemodialysis vascular access and endovascular procedures. American Society of Diagnostic and Interventional Nephrology. PMID- 12641885 TI - The technical and interpersonal aspects of telemedicine: effects on doctor patient communication. AB - The influence of telemedicine on the nature and content of doctor-patient communication stems from both its technical and its interpersonal aspects. While the technical aspects are concerned with the communication technologies used and the clinical processes enabled by those technologies, the interpersonal aspects are concerned with relationships between system personnel, providers and patients, and the way in which those relationships are organized. On the one hand, this paper posits that the influence of the technical environment stems from depersonalization of the doctor-patient relationship, participatory enhancements and impediments, and sensory and non-verbal limitations. On the other hand, it posits that the influence of the interpersonal environment stems from third-party participation, social and professional distancing, and underdeveloped norms and standards. A combined positivist and interpretivist evaluation strategy would enable researchers to make better-informed connections between telemedicine, medical encounter behaviour and health outcomes. PMID- 12641886 TI - Videoconferencing for orthopaedic outpatients: one-year follow-up. AB - We studied whether consultations via videoconferencing and traditional outpatient clinic visits differ in terms of the implementation of the patient management plan during a one-year follow-up. First-admission and follow-up orthopaedic patients were randomly allocated to an outpatient visit at the surgical department of Oulu University Hospital or to videoconferencing at a health centre in Pyhajarvi. In a prospective one-year study, there were 145 consecutive orthopaedic patients who met the inclusion criteria: 84 referred for their first visit to a specialist and 61 of them for follow-up. There were 66 males (46%) in the study population. Over half the patients had some form of regenerative arthritis: 15% had hip arthritis, 33% knee arthritis and 4% other arthritis. There were no differences in the implementation of the management plan between the two groups. The study showed that videoconferencing is a valid alternative to outpatient clinic visits for orthopaedic specialist consultations. PMID- 12641887 TI - The effectiveness of telepsychiatry measured using the Health of the Nation Outcome Scale and the Mental Health Inventory. AB - In a telepsychiatry project in rural Queensland, data were collected from 124 patients attending hospital and general practice facilities for mental health care and then again at follow-up one year later. Thirty-two of the patients were dealt with using telepsychiatry. Two health status scales were used to measure effectiveness: the Health of the Nation Outcome Scale (HoNOS), administered by the practitioners; and the Mental Health Inventory (MHI), which was self administered by the patients. There was a significant difference between the initial assessment and follow-up groups on most subscales of the HoNOS, but no significant difference between the face-to-face and telepsychiatry groups. Similarly, the MHI results showed a significant difference on all subscales between the initial assessment and follow-up groups, but no significant difference between the face-to-face and telepsychiatry groups. Individuals who used and did not use telepsychiatry all had improved health outcome scores on the HoNOS and MHI during the study period. Telepsychiatry was as effective as face-to face care. PMID- 12641888 TI - A comparison between three electronic media and in-person learning for continuing education in physical rehabilitation. AB - We produced continuing education material in physical rehabilitation using a variety of electronic media. We compared four methods of delivering the learning modules: in person with a computer projector, desktop videoconferencing, Web pages and CD-ROM. Health-care workers at eight community hospitals and two nursing homes were asked to participate in the project. A total of 394 questionnaires were received for all modalities: 73 for in-person sessions, 50 for desktop conferencing, 227 for Web pages and 44 for CD-ROM. This represents a 100% response rate from the in-person, desktop conferencing and CD-ROM groups; the response rate for the Web group is unknown, since the questionnaires were completed online. Almost all participants found the modules to be helpful in their work. The CD-ROM group gave significantly higher ratings than the Web page group, although all four learning modalities received high ratings. A combination of all four modalities would be required to provide the best possible learning opportunity. PMID- 12641889 TI - Telesurveillance of elderly patients by use of passive infra-red sensors in a 'smart' room. AB - We have developed an automated surveillance system based on passive infra-red sensors. Eight sensors were installed in a hospital room. A computer automatically captured data from the sensors every night from 21:00 until 06:00 the following morning. The sensors were polled twice per second and when a sensor was activated by movement, the event and time were recorded in a data file. At the end of the surveillance period the program analysed the data and generated a report showing the activities taking place in the room and their times. Four elderly patients were observed for a total of 97 nights. A total of 1637 possible sequences of movements by the patient and the hospital staff were detected. The computer was able to identify 1450 sequences (89%) correctly, in comparison with manual analysis. Only 10 movements (0.6%) were undetected by the system; all were very short sequences (five or six activations of the sensors). The system was generally capable of detecting and classifying all major movements in the room. PMID- 12641890 TI - Peer support via video-telephony among frail elderly people living at home. AB - We evaluated the effect of a video-phone network on peer support among frail elderly people living at home. We conducted a one-year trial of network formation among 14 people (five men and nine women, age range 78-85 years) through coordinator intervention and the use of ISDN video-phones. The purpose of the intervention was to support and improve the functional independence of frail elderly people at home and to widen their social network. During the study, the subjects made 1400 video-calls, lasting a total of 25,867 min. One subject made no calls. There was greater use of video-phones by those living outside the central city area. Calls between men and women were infrequent. The men generally had shorter call times and made fewer calls. The participants could be categorized into three groups: those using video-phones every day; those using them once per week; and those using them once per month. All subjects in the first group clearly expressed their satisfaction with the video-phone. A video phone network appears to be helpful for elderly people in their peer support relationships. PMID- 12641891 TI - The quality of medical records in teleconsultation. AB - We collected and examined the medical records from telemedicine cases dealt with by the telemedicine centre of Shanghai Hospital No. 85. This centre handles the second largest number of teleconsultations in the entire network. There were 658 telemedicine cases in total. The medical records included the patient record in 599 cases (91%), transmitted images in 392 cases (60%), the consultant's opinion in 595 cases (90%) and a video-recording of the teleconsultation in 203 cases (31%). The quality of patient records was reviewed and found to be acceptable in 58% of cases. In total, 1794 radiology images (85% of all images) were transmitted via the telemedicine network. The consultant considered 352 of them (20%) to be unreadable on the screen (i.e. 80% of radiology images were considered to be acceptable). For optimum performance of telemedicine, the patient record and associated images should be delivered in advance and the relevant parts of the patient record should be available during a teleconsultation. Three aspects of the management of the medical records for teleconsultations are particularly important: multimedia collection, standardization of patient/record identification and classification, and information management. PMID- 12641892 TI - Patient satisfaction with teledermatology services. AB - We collected data on patient satisfaction with the use of teledermatology services. During a 27-month study, a single dermatologist saw a total of 321 patients via telemedicine. The patients completed 483 surveys, although not all questions were answered on all surveys. There was a high level of satisfaction among patients using teledermatology: 88% of 258 respondents said that they were satisfied with their telemedicine session. In terms of the eight questions on the questionnaire, 84% of the responses were in the top two positive categories on a seven-point Likert scale. In addition, access to dermatology appeared to increase with the availability of the telemedicine service. PMID- 12641893 TI - Development of the Telemedicine Satisfaction Questionnaire to evaluate patient satisfaction with telemedicine: a preliminary study. AB - We developed a questionnaire for the evaluation of patient satisfaction with telemedicine. Items related to patient satisfaction with telemedicine were identified through a review of the literature. The internal consistency and the intraclass correlation coefficient were used to evaluate the reliability of the proposed instrument. Content validity was examined using a panel of experts. The predictive validity of the proposed instrument was assessed by correlating scores on the Telemedicine Satisfaction Questionnaire (TSQ) with haemoglobin A(1c) levels and adherence to self-management activities among 38 patients with type 2 diabetes, after they had completed a four-session interactive diabetes tele education programme. A principal-components analysis with varimax rotation was performed to reduce the number of questionnaire items from 15 to 14, and the scree test was used to select significant factors. There were three components with eigenvalues over 1.0, which together explained 68% of the total variance. These were: quality of care provided, similarity to face-to-face encounter and perception of the interaction. The TSQ demonstrated preliminary reliability and validity but more extensive testing will be required before it can be considered generally applicable. PMID- 12641894 TI - An exploratory study of language interpretation services provided by videoconferencing. AB - We explored the feasibility and acceptability of the remote provision of a language interpretation service during general practice consultations. Three methods were used to provide an interpretation service: a physically present interpreter (PI), a remote interpreter accessed using an ISDN videoconferencing link at 128 kbit/s (VI) and a remote interpreter accessed by telephone (TI). Thirty-six non-English-speaking patients were invited to take part and 35 agreed to do so. Twenty-nine (83%) of the patients recruited were female. The age range of the participants was 24-51 years. Fourteen consultations took place with a physically present interpreter, 11 using videoconferencing and 10 using the telephone. Mean scores on the Patient Enablement Instrument were 5.2 for the PI group, 2.3 for the VI group and 5.1 for the TI group. Mean scores on the Medical Interview Satisfaction Scale were 5.3 for the PI group, 4.9 for the VI group and 5.3 for the TI group. The visual quality and sound quality of remote interpreting were satisfactory. Both videoconferencing and hands-free telephones can deliver an acceptable interpreting service in primary care. PMID- 12641895 TI - Effect of videoconferencing on the process of cooperation between primary and secondary care. PMID- 12641896 TI - The use of videoconferencing to enhance student learning. PMID- 12641897 TI - Seduced by technology? PMID- 12641898 TI - Genital morphology and fertilization success in the dung beetle Onthophagus taurus: an example of sexually selected male genitalia. AB - In animals with internal fertilization and promiscuous mating, male genitalia show rapid and divergent evolution. Three hypotheses have been suggested to explain the evolutionary processes responsible for genital evolution: the lock and-key hypothesis, the pleiotropy hypothesis and the sexual-selection hypothesis. Here, we determine whether variation in male genital morphology influences fertilization success in the dung beetle Onthophagus taurus, as predicted by the sexual-selection hypothesis. Variation in four out of five genital sclerites of the endophallus influenced a male's fertilization success, supporting the general hypothesis that male genitalia can evolve under sexual selection. Furthermore, different genital sclerites were found to enhance first versus second male paternity, indicating that different sclerites serve offensive and defensive roles. Genital-trait variability was comparable to that in other species but was less variable than a non-genital sexually selected trait (head horns). We suggest that directional selection for genital elaboration may be countered by natural selection, which should favour genitalia of a size and shape necessary for efficient coupling and sperm transfer. PMID- 12641899 TI - How the global structure of protein interaction networks evolves. AB - Two processes can influence the evolution of protein interaction networks: addition and elimination of interactions between proteins, and gene duplications increasing the number of proteins and interactions. The rates of these processes can be estimated from available Saccharomyces cerevisiae genome data and are sufficiently high to affect network structure on short time-scales. For instance, more than 100 interactions may be added to the yeast network every million years, a fraction of which adds previously unconnected proteins to the network. Highly connected proteins show a greater rate of interaction turnover than proteins with few interactions. From these observations one can explain (without natural selection on global network structure) the evolutionary sustenance of the most prominent network feature, the distribution of the frequency P(d) of proteins with d neighbours, which is broad-tailed and consistent with a power law, that is: P(d) proportional, variant d (-gamma). PMID- 12641900 TI - Climate change and habitat destruction: a deadly anthropogenic cocktail. AB - Climate change and habitat destruction are two of the greatest threats to global biodiversity. Lattice models have been used to investigate how hypothetical species with different characteristics respond to habitat loss. The main result shows that a sharp threshold in habitat availability exists below which a species rapidly becomes extinct. Here, a similar modelling approach is taken to establish what determines how species respond to climate change. A similar threshold exists for the rate of climate change as has been observed for habitat loss-patch occupancy remains high up to a critical rate of climate change, beyond which species extinction becomes likely. Habitat specialists, especially those of relatively poor colonizing ability are least able to keep pace with climate change. The interaction between climate change and habitat loss might be disastrous. During climate change, the habitat threshold occurs sooner. Similarly, species suffer more from climate change in a fragmented habitat. PMID- 12641901 TI - Climate tolerance and interspecific variation in geographic range size. AB - The fact that climate influences the continental-scale distributions of species is one of the central tenets of ecology and biogeography. Equally elemental is that species exhibit enormous variation in geographic range size, with most occupying comparatively small areas. The degree to which climate can account for this variation remains unclear. Here, I test three alternative climate-based hypotheses for variation in range size using a large sample of tree and shrub species native to North America north of Mexico. I show that the lowest value of January average daily minimum temperature encompassed by a species' geographic range (T(MIN)), representing the 'climate extremes hypothesis', explains almost 80% of the variation in range size. Hypotheses based on seasonality and climate optima find substantially less support. The relationship between range size and T(MIN) does not change across the breadth of latitudes examined, and is general for conifers and hardwoods, and growth form (tree versus shrub). Differential freezing resistance gains support as the mechanism underlying interspecific variation in range size: using 35 species for which data were available, both T(MIN) and range size are shown to be strongly correlated with measures of freezing resistance. PMID- 12641902 TI - Stroke and glide of wing-propelled divers: deep diving seabirds adjust surge frequency to buoyancy change with depth. AB - In order to increase locomotor efficiency, breath-holding divers are expected to adjust their forward thrusts in relation to changes of buoyancy with depth. Wing propulsion during deep diving by Brunnich's guillemots (Uria lomvia) was measured in the wild by high-speed (32 Hz) sampling of surge (tail-to-head) and heave (ventral-to-dorsal) accelerations with bird-borne data loggers. At the start of descent, the birds produced frequent surges (3.2 Hz) during both the upstroke and the downstroke against buoyancy to attain a mean speed of 1.2-1.8 m s(-1) that was close to the expected optimal swim speed. As they descended deeper, the birds decreased the frequency of surges to 2.4 Hz, relaying only on the downstroke. During their ascent, they stopped stroking at 18 m depth, after which the swim speed increased to 2.3 m s(-1), possibly because of increasing buoyancy as air volumes expanded. This smooth change of surge frequency was achieved while maintaining a constant stroke duration (0.4-0.5 s), presumably allowing efficient muscle contraction. PMID- 12641903 TI - Model of a predatory stealth behaviour camouflaging motion. AB - A computational model of a stealth strategy inspired by the apparent mating tactics of male hoverflies is presented. The stealth strategy (motion camouflage) paradoxically allows a predator to approach a moving prey in such a way that it appears to be a stationary object. In the model, the predators are controlled by neural sensorimotor systems that base their decisions on realistic levels of input information. They are shown to be able to employ motion camouflage to approach prey that move along both real hoverfly flight paths and artificially generated flight paths. The camouflaged approaches made demonstrate that the control systems have an ability to predict future prey movements. This is illustrated using two- and three-dimensional simulations. PMID- 12641904 TI - Contrasting relatedness patterns in bottlenose dolphins (Tursiops sp.) with different alliance strategies. AB - Male bottlenose dolphins (Tursiops sp.) in Shark Bay have one of the most complex male societies outside humans. Two broad mating strategies have been identified in males. In the first strategy, there are two types of alliances: stable 'first order' pairs and trios that herd individual females in reproductive condition, and 'second-order' teams of two first-order alliances (five or six individuals) that join forces against rivals in contests for females. In the alternative strategy, a 'super-alliance' of ca. 14 individuals, males form pairs or trios to herd females, but in contrast to the stable alliances, these pairs and trios are highly labile. Here, we show that males in stable first-order alliances and the derived second-order alliances are often strongly related, so that they may gain inclusive fitness benefits from alliance membership. By contrast, members of the super-alliance are no more closely related than expected by chance. Further, the strength of the association of alliance partners within the super-alliance, as measured by an index of joint participation in consorting a female, was not correlated with their genetic relatedness. Thus, within one population and one sex, it appears that there may be simultaneous operation of more than one mode of group formation. PMID- 12641906 TI - Information transfer about roosts in female Bechstein's bats: an experimental field study. AB - Information transfer among group members is believed to play an important part in the evolution of coloniality in both birds and bats. Although information transfer has received much scientific interest, field studies using experiments to test the underlying hypotheses are rare. We used a field experiment to test if communally breeding female Bechstein's bats (Myotis bechsteinii) exchange information regarding novel roosts. We supplied a wild colony, comprising 17 adult females of known relatedness, with pairs of suitable and unsuitable roosts and monitored the arrival of individuals marked with transponders (PIT-tags) over 2 years. As expected with information transfer, significantly more naive females were recruited towards suitable than towards unsuitable roosts. We conclude that information transfer about roosts has two functions: (i) it generates communal knowledge of a large set of roosts; and (ii) it aids avoidance of colony fission during roost switching. Both functions seem important in Bechstein's bats, in which colonies depend on many day roosts and where colony members live together for many years. PMID- 12641905 TI - Wild female baboons bias their social behaviour towards paternal half-sisters. AB - Adult female cercopithecines have long been known to bias their social behaviour towards close maternal kin. However, much less is understood about the behaviour of paternal kin, especially in wild populations. Here, we show that wild adult female baboons bias their affiliative behaviour towards their adult paternal half sisters in the same manner and to the same extent that they bias their behaviour towards adult maternal half-sisters. Females appear to rely heavily on social familiarity as a means of biasing their behaviour towards paternal half-sisters, but may use phenotype matching as well. PMID- 12641907 TI - Pollinator attraction in a sexually deceptive orchid by means of unconventional chemicals. AB - Ophrys flowers mimic virgin females of their pollinators, and thereby attract males for pollination. Stimulated by scent, the males attempt to copulate with flower labella and thereby ensure pollination. Here, we show for the first time, to our knowledge, that pollinator attraction in sexually deceptive orchids may be based on a few specific chemical compounds. Ophrys speculum flowers produce many volatiles, including trace amounts of (omega-1)-hydroxy and (omega-1)-oxo acids, especially 9-hydroxydecanoic acid. These compounds, which are novel in plants, prove to be the major components of the female sex pheromone in the scoliid wasp Campsoscolia ciliata, and stimulate male copulatory behaviour in this pollinator species. The specificity of the signal depends primarily on the structure and enantiomeric composition of the oxygenated acids, which is the same in wasps and in the orchids. The overall composition of the blend differs significantly between the orchid and its pollinator and is of secondary importance. 9 Hydroxydecanoic acid is a rarely occurring compound that until now has been identified only in honeybees. Contrary to the standard hypothesis that Ophrys flowers produce only 'second-class attractivity compounds' and are neglected once the pollinator females are present, we show that flowers are more attractive to the males than are their own females. PMID- 12641908 TI - Influence of female reproductive anatomy on the outcome of sperm competition in Drosophila melanogaster. AB - Females as well as males can influence the outcome of sperm competition, and may do so through the anatomy of their reproductive tracts. Female Drosophila melanogaster store sperm in two morphologically distinct organs: a single seminal receptacle and, normally, two spermathecae. These organs have different temporal roles in sperm storage. To examine the association between sperm storage organ morphology and sperm competition, we used a mutant type of female with three spermathecae. Although the common measure of sperm competition, P(2), did not differ between females with two and three spermathecae, the pattern of sperm use over time indicated that female morphology did affect male reproductive success. The rate of offspring production by females with three spermathecae rose and fell more rapidly than by females with two spermathecae. If females remate or die before using up second male sperm, then second male reproductive success will be higher when they mate with females with three spermathecae. The results indicate that temporal patterns of sperm use as well as P(2) should be taken into account when measuring the outcome of sperm competition. PMID- 12641909 TI - Differential investment and costs during avian incubation determined by individual quality: an experimental study of the common eider (Somateria mollissima). AB - Individuals of different quality may have different investment strategies, shaping responses to experimental manipulations, thereby rendering the detection of such patterns difficult. However, previous clutch-size manipulation studies have infrequently incorporated individual differences in quality. To examine costs of incubation and reproductive investment in relation to changes in clutch size, we enlarged and reduced natural clutch sizes of four and five eggs by one egg early in the incubation period in female common eiders (Somateria mollissima), a sea duck with an anorectic incubation period. Females that had produced four eggs (lower quality) responded to clutch reductions by deserting the nest more frequently but did not increase incubation effort in response to clutch enlargement, at the cost of reduced hatch success of eggs. Among birds with an original clutch size of five (higher quality), reducing and enlarging clutch size reduced and increased relative body mass loss respectively without affecting hatch success. In common eiders many females abandon their own ducklings to the care of other females. Enlarging five-egg clutches led to increased brood care rate despite the higher effort spent incubating these clutches, indicating that the higher fitness value of a large brood is increasing adult brood investment. This study shows that the ability to respond to clutch size manipulations depends on original clutch size, reflecting differences in female quality. Females of low quality were reluctant to increase investment at the cost of lower hatch success, whereas females of higher quality apparently have a larger capacity both to increase incubation effort and brood care investment. PMID- 12641910 TI - Evolutionary coherence of the mammalian amygdala. AB - Despite great interest in the role of the amygdala in animal and human behaviour, its very existence as a structurally and functionally unified brain component has been questioned, on the grounds that cell groups within it display divergent pharmacological and connectional characteristics. We argue that the question of whether particular brain nuclei constitute a valid structural and functional unit is inherently an evolutionary question, and we present a method for answering it. The method involves phylogenetic analysis of comparative data to determine whether or not separate regions of the putative brain structure show statistically correlated evolution. We find that, in three separate groups of mammals (primates and two groups of insectivores), evolutionary changes in the volumes of amygdala components are strongly correlated, even after controlling for volumetric change in a wide range of limbic and other brain structures. This allows us to reject the strong claim that the amygdala is neither a structural nor a functional unit, and demonstrates the importance of evolutionary analysis in resolving such issues in systems neuroscience. PMID- 12641912 TI - [Cardiology]. PMID- 12641911 TI - The de novo selection of drug-resistant malaria parasites. AB - Antimalarial drug resistance emerges de novo predominantly in areas of low malaria transmission. Because of the logarithmic distribution of parasite numbers in human malaria infections, inadequately treated high biomass infections are a major source of de novo antimalarial resistance, whereas use of antimalarial prophylaxis provides a low resistance selection risk. Slowly eliminated antimalarials encourage resistance largely by providing a selective filter for resistant parasites acquired from others, and not by selecting resistance de novo. The de novo emergence of resistance can be prevented by use of antimalarial combinations. Artemisinin derivative combinations are particularly effective. Ensuring adequate treatment of the relatively few heavily infected patients would slow the emergence of resistance. PMID- 12641913 TI - [Respiratology]. PMID- 12641914 TI - [Nephrology]. PMID- 12641915 TI - [Gastroenterology]. PMID- 12641916 TI - [Hematology]. PMID- 12641917 TI - [Rheumatology]. PMID- 12641919 TI - [Psychiatry]. PMID- 12641918 TI - [Neurology]. PMID- 12641920 TI - [Endocrinology]. PMID- 12641922 TI - [Parasitology]. PMID- 12641921 TI - [Lemology]. PMID- 12641923 TI - [Tuberculosis]. PMID- 12641924 TI - [Gerontology]. PMID- 12641925 TI - [Endemiology]. PMID- 12641926 TI - [Cardiovascular surgery]. PMID- 12641927 TI - [General surgery]. PMID- 12641928 TI - [Urology]. PMID- 12641929 TI - [Neurosurgery]. PMID- 12641930 TI - [Organ transplantation]. PMID- 12641931 TI - [Otorhinolaryngology-head and neck surgery]. PMID- 12641932 TI - [Orthognathic surgery]. PMID- 12641933 TI - [Traumatology]. PMID- 12641934 TI - [Gynecology]. PMID- 12641935 TI - [Obstetrics]. PMID- 12641936 TI - [Pediatrics]. PMID- 12641937 TI - [Sexually transmitted diseases]. PMID- 12641938 TI - [Ultrasonic medicine]. PMID- 12641939 TI - [Radiology]. PMID- 12641941 TI - [Laboratory medicine]. PMID- 12641940 TI - [Radiological medicine and protection]. PMID- 12641942 TI - [Nuclear medicine]. PMID- 12641943 TI - [Pathology]. PMID- 12641944 TI - [Cytobiology]. PMID- 12641945 TI - [Pharmacology]. PMID- 12641946 TI - [Chinese traditional medicine and pharmacy]. PMID- 12641947 TI - [Physical medicine and rehabilitation]. PMID- 12641948 TI - [Chemosensitivity testing of oral and maxillofacial cancer with biopsy specimens]. AB - OBJECTIVE: To determine the chemosensitivity in fresh biopsy specimen of human oral and maxillofacial cancer, and the differential chemosensitivity among those drugs used popularly in clinic. METHODS: Human biopsy cancer cells were obtained from 150 oral and maxillofacial malignant tumors. The antitumor drugs tested using modified MTT assay were cisplatin (CDDP), 5-fluorouracil (5-Fu), Pinyangmycin (PYM), Paclitaxel (Taxol), Teniposide (Vm-26), Epi-adriamycin (E ADM), Vindesin (VDS) and Methortrexatum (MTX). RESULTS: The success rate of the MTT assay was 93.33% (140 of the 150 cases). At a drug concentration of Cmax x 5, the inhibition rates of oral tumor cells were 63.76% for Vm-26, 25.93% for CDDP, 25.86% for E-ADM, 23.52% for Taxol, 22.97% for PYM, 22.08% for 5-Fu, 18.42% for VDS and 18.93% for MTX. The inhibition rate of VM26 was significantly higher than any of other seven chemotherapeutic drugs (P < 0.05). Over forty percent patients with squamous cell carcinoma showed moderate chemosensitivity to VM-26, CDDP and E-ADM, and over forty percent cases with adenoid carcinoma showed moderate chemosensitivity to Vm-26, Taxol and E-ADM. CONCLUSIONS: Most oral and maxillofacial cancers showed chemosensitivity to Vm-26, CDDP, E-ADM and Taxol. Vm 26, E-ADM and Taxol were more potent drugs than VDS, 5-Fu and MTX against oral and maxillofacial cancer cells. Chemosensitivity testing using modified MTT assay was useful in selecting antitumor drugs for patients with oral and maxillofacial cancers. PMID- 12641949 TI - [Chromosome localization of the dentinogenesis imperfecta type II locus]. AB - OBJECTIVE: To investigate the linkage between dentinogenesis imperfecta type II and chromosome 4q21 in a Tianjin-Tanggu family of the Hui nationality. METHODS: Blood samples were collected from 13 family members. DNAs were analyzed with 8 STRP markers (D4S2915, D4S2932, GATA62A11, D4S2409, DSP STRP, SPP1 STRP, D4S1563, D4S1544) using fluorescence-based PCR. The linkage between eight markers on chromosome 4q21 and dentinogenesis imperfecta type II locus was tested respectively by lod score analysis. RESULTS: Genotype and haplotype were acquired. Genetic linkage analysis demonstrated the maximum lod score of eight STRPs were all larger than zero, in which five of them were larger than 1. CONCLUSION: The locus of dentinogenesis imperfecta type II in Chinese family is located on human chromosome 4q21, which indicated that the locus of Chinese Hui nationality should be the same as that of other reported European or American family. PMID- 12641950 TI - [A clinical and histological study of Bio-Oss in combination with guided bone regeneration membrane technique]. AB - OBJECTIVE: To evaluate the reliability of Bio-Oss particles as bone grafts in combination with membrane-technique. METHODS: 6 small biopsy specimens were retrieved without causing iatrogenic damage in 6 patients (implant sites). The Donath's hard tissue section and micro grinding technique was adopted, and Leval Laczko staining method was used. RESULTS: The histological results showed Bio-Oss particles were easily distinguished from the newly formed bone, but new bone was formed among different particles of Bio-Oss, no connective tissue was found. Intimate contact between newly formed bone and particles of the graft was present. CONCLUSIONS: Bio-Oss appears to be highly biocompatible and osteoconductive. It can be used success fully as a bone substitute in combination with guided bone regeneration membrane technique. PMID- 12641951 TI - [The lingual region upper airway CT scan of obstructive sleep apnea patients]. AB - OBJECTIVE: To compare the lingual region upper airway CT scan results of OSAS patients and normal adults to explore if there are upper airway stricture in OSAS patients and the reason of stricture. METHODS: 59 patients with OSAS and 57 normal adults were included in the study. CT evaluated the lingual region upper airway with the use of a Philips Tomoscan AV Expander E1 spiral scanner. The shapes of lingual region upper airway were observed. The area and the dimensions of lingual region was studied, and the thickness of retropharyngeal and lateral pharyngeal tissue were evaluated too. The another focus area was tongue, and the evaluated data included the tongue width, genioglossus width, hyogolssus width, tongue length and tongue area. RESULTS: The data comparative results of OSAS and normal groups were as follows: the right to left diameter and area of OSAS patients' lingual region upper airways were less than those of normal adults, the thickness of lateral pharyngeal, tongue width, genioglossus width, hyogolssus width and tongue area were more than those of normal adults. And there were not obviously difference between OSAS patients and normal adults in tongue length, anterior-to-posterior dimension and thickness of retropharyngeal wall tissue of upper airway. CONCLUSIONS: The study suggests that there are obviously difference between the patients with OSAS and normal adult in lingual upper airway CT scan measure. And the anatomic stricture of lingual upper airway is one of the etiology of OSAS. The main reasons of stricture are increasing of thickness of lateral pharyngeal, tongue width, genioglossus width, hyogolssus width and tongue area. PMID- 12641952 TI - [Evaluation for submental island flap]. AB - OBJECTIVE: To evaluate the clinical effect of submental island flap. METHODS: The influence of head and neck radiotherapy and neck lymph node metastasis on the flap was analysed. The clinical effect in the near future and longterm of submental island flap for defects repair was evaluated. RESULTS: Thirty-eight submental island flap were applied for reconstruction of head and neck defects following cancer ablation between July of 1997 to December of 1999. 16 of these had preoperative radiotherapy at both donor site and receive site, and neck dissections with resection of primary lesions were simultaneously performed in 18 patients. The success rate was 89.5% (34 of 38). All repaired cases had well matched skin with facial surface and excellent texture for intraoral function. CONCLUSIONS: Routine preoperative radiation does do not influence the success and healing of the flap. For tumor complete resection, appropriate cases should be chosen. We believe this flap is worthwhile in addition to the existing armamentarium of reconstruction options both facially and intraorally. PMID- 12641953 TI - [Quantitative study of the cellular nuclear DNA contents in odontogenic cysts and ameloblastoma]. AB - OBJECTIVE: To investigate the proliferative potential of the epithelial cells in odontogenic keratocyst, radicular cyst, dentigerous cyst and ameloblastoma. METHODS: DNA contents and ploidy of basal and spinous cells in keratocyst, radicular cyst, dentigerous cyst, and the peripheral column cells and central reticular cells in ameloblastoma were analysis respectively. RESULTS: The more and higher DNA contents and the proliferating ploidy of keratocyst and ameloblastoma than those of radicular cyst and dentigerous cyst indicate the active proliferating potential. The spinous cells showed more active proliferating growth than the basal cells of keratocyst. The higher DNA contents of radicular cyst are related to the stimulus of the inflammation. The dentigerous cysts have more di-ploidy cells without active growth potential. CONCLUSIONS: The active cell proliferating growth in keratocyst and ameloblastoma is probably the pathological basis of their local aggressive biological behavior. PMID- 12641954 TI - [A histological and ultrastructural study of the tidemark in human condylar cartilage]. AB - OBJECTIVE: To study the histology and ultrastructure of the tidemark in the adult condylar cartilage and their significance. METHODS: After embedded in paraffin, 50 adult condyles were stained with HE, partly with Van-Gieson and histochemical methods, then observed by light microscope. 3 cases of the tidemark region were observed by transmission electron microscope, another 5 cases were studied by scanning electron microscope. RESULTS: In the tidemark region, there had shown the presence of the AKP and calcium, absence of the proteoglycan, abundance of the membrane-bound matrix vesicles, crystals of hydroxyapatite and lipid nodule like substances, which were often observed in the load-bearing areas. The collagen fibrils of the noncalcified cartilage crossed the tidemark gradiently and were continuous with those of the calcified cartilage; a band of horizontal fibrils surrounded the whole tidemark region, which was wider in the load-bearing areas than that in the nonload-bearing areas and which interweaved with the gradient fibrils so as to form a net. Digested with papain, the surface of the tidemark was highly undulating, and a lot of chondrocyte lacunae were seen on the surface, which were surrounded by calcified tissues. CONCLUSIONS: In the region of the tidemark, physiological calcification takes place and is more active in the load-bearing areas; gradient and horizontal fibrils interweave with each other, which is correlated with the force on the articulation. PMID- 12641955 TI - [Effect of jaw shape on stresses at implant-bone interface: a three-dimensional finite element analysis]. AB - OBJECTIVE: To study the influence of different shape of lower jaw on stress distribution at bone interface in the mandibular molar region of implant restoration models. METHODS: Six models with different lower jaw shapes were constructed by using three-dimensional finite element method. Implant-bone interface stresses in these models were studied under vertical and lateral loads. RESULTS: No matter in the condition of vertical loading of lateral loading, stress distribution patterns were similar in variant models and differences of stress values were not statistically significant. The maximal differences in Von Mises stress, compressive stress and tensile stress among the six models were 6.4%, 2.8% and 6.2%respectively. CONCLUSION: It is rational to simplify the lower jaw shape in finite element analysis. PMID- 12641956 TI - [Electromyography activities of the head, neck and upper trunk muscles with mandibular movement in normal adults]. AB - OBJECTIVE: To investigate the activities of head, neck and upper trunk muscles with mandibular movement in normal adults. METHODS: The integral data of surface electromyography (EMG) had been recorded to analyze the activities of anterior temporal (Ta), posterior temporal (Tp), sternocleidomastoid (SCM) and trapezius (TRAP) muscles with mandibular movement in ten normal adults. RESULTS: The former mentioned muscles acted constantly when the mandible was in the rest position. The activities of Ta, Tp and SCM muscles increased with protrusion of mandible, mouth opening, tapping, maximum clenching, and chewing movements. The amplitudes of the increased activity of these muscles were correlated significantly with each other. When the activities of Ta and Tp muscles increased to 56 times and 25 times respectively, the activity of SCM muscle increased to 5 - 6 times. The activity of trapezius muscle increased with protrusion of mandible and opening movement, but tended to stop with the maximum clenching and chewing movement. The correlations of left and right sides of the same muscles had been found. CONCLUSIONS: The muscles of head, neck and upper trunk can act as contributors to keep the mandibular positions and take part in the mandibular movements. PMID- 12641957 TI - [A study on safety of opening pulp chamber therapy for patients with a history of remote myocardial infarction]. AB - OBJECTIVE: To study the indication and contraindication of opening pulp chamber therapy for patients with a history of remote myocardial infarction. METHODS: Blood pressure (BP), heart beat rate and electrocardiographic examination were determined during opening pulp chamber therapy in patients with acute and chronic pulpititis and a history of remote myocardial infarction. RESULTS: High blood pressure and quick heart beat were found during local anaesthetic injection and opening of pulp chamber (during local anaesthetic injection BP increased by 12.52/8.04 mmHg, heart rate by 4.02 times/min;during opening of pulp chamber BP and heart rate increased by 15.43/8.0 mmHg, 6.37 times/min respectively). Electrocardiogram showed myocardial ischemic pattern deteriorated in some patients with the history of coronary insufficiency. CONCLUSIONS: Most patients can bear it. Comprehensive physical examinations, evaluations of the patients' cardial status, mental state and indispensible equipment are necessary. The therapy is recommended to be operated under the monitor electrocardiography for the safety of the patients. PMID- 12641958 TI - [Comparison of machinability of two types of dental machinable ceramic]. AB - OBJECTIVE: In terms of the problems of now available dental machinable ceramics, a new type of calcium-mica glass-ceramic, PMC-I ceramic, was developed, and its machinability was compared with that of Vita MKII quantitatively. Moreover, the relationship between the strength and the machinability of PMC-I ceramic was studied. METHODS: Samples of PMC-I ceramic were divided into four groups according to their nucleation procedures. 600-seconds drilling tests were conducted with high-speed steel tools (Phi = 2.3 mm) to measure the drilling depths of Vita MKII ceramic and PMC-I ceramic, while constant drilling speed of 600 rpm and constant axial load of 39.2 N were used. And the 3-point bending strength of the four groups of PMC-I ceramic were recorded. RESULTS: Drilling depth of Vita MKII was 0.71 mm, while the depths of the four groups of PMC-I ceramic were 0.88 mm, 1.40 mm, 0.40 mm and 0.90 mm, respectively. Group B of PMC I ceramic showed the largest depth of 1.40 mm and was statistically different from other groups and Vita MKII. And the strength of the four groups of PMC-I ceramic were 137.7, 210.2, 118.0 and 106.0 MPa, respectively. CONCLUSION: The machinability of the new developed dental machinable ceramic of PMC-I could meet the need of the clinic. PMID- 12641959 TI - [Pre-surgical orthodontic treatment of skeletal class II patients with gingival smile corrected by anterior maxillary segmental osteotomy]. AB - OBJECTIVE: To discuss the pre-surgical orthodontic treatment of skeletal class II patients with gingiva smile corrected by AMSO. METHODS: We analyzed the clinical features of 20 skeletal class II patients treated by AMSO combined with Orthodontic treatment and evaluated the effects of AMSO by means of cephalometric analysis. RESULTS: After the AMSO treatment, ANB angle, the height of anterior maxilla, the protrusion of the upper anterior teeth, and the of A point had reduced significantly (P < 0.05). After AMSO, the appearance of patients had been improved markedly. CONCLUSIONS: AMSO can correct the protrusion of the maxilla and gingival smie efficiently. The anchorage of molars should be controlled carefully. The anterior part of the upper arch should be expended orthodontically to make arch relationship. Extract the upper bicuspid half a year before the surgery was recommended. When necessary, genioplasty could be performed. PMID- 12641960 TI - [Effects of different class II cavity designs on stresses in restoration]. AB - OBJECTIVE: To assess effects of different class II cavities on stresses of amalgam and composite resin restoration. METHODS: Five kinds of class II MO cavities were prepared on the first molars of mandibles. The cavities were filled by amalgam and composite resin, respectively. The stresses were analyzed by using three-dimensional finite element method, after vertical and lateral pressures were loaded on the teeth. RESULTS: The stresses were greater in traditional cavities than those in reformed cavities when 100 N vertical and lateral pressures were loaded on the teeth. But the pressing stress and shearing stress in cavities filled by amalgam were smaller in traditional cavities than those in reformed cavities. The stresses were smaller in cavities filled by resin than those in cavities filled by amalgam significantly, when the cavities were designed with slot retention groove, without slot retention groove and as plate form. The stresses were smaller in two kinds of slot and plate cavities than those in traditional and reformed cavities. CONCLUSION: It is better to design cavity with slot retention groove for amalgam restoration and to design cavity without slot retention groove for composite resin restoration. PMID- 12641961 TI - [Detection of hepatitis C virus antibody in saliva and its clinical significance]. AB - OBJECTIVE: To detect hepatitis C virus (HCV) antibody in saliva and find its clinical significance. METHODS: Sera and saliva samples from eighty-six patients were parallely detected HCV antibodies (anti-HCV) with EIA using a modified protocol. RESULTS: Twenty-nine of 86 patients were positive for anti-HCV in sera samples; and, 27/29 were also anti-HCV positive with saliva samples. For the 57 anti-HCV negative sera, the saliva samples were all negative. Comparing with the results of sera, the sensitivity (93.1%, 27/29) and specificity (100%, 57/57) of salivary detection for anti-HCV could be accepted. CONCLUSION: If the results of sera are considered as "golden standard", these results suggest that tests on saliva can be useful in HCV clinical diagnosis and epidemiological studies for estimating the prevalence of HCV in populations. PMID- 12641962 TI - [Intranasal immunization against dental caries with plasmid DNA encoding pac gene of Streptococcus mutans in gnotobiotic rats]. AB - OBJECTIVES: To assess the efficacy of plasmid DNA encoding pac gene of Streptococcus mutans (S. mutans) intranasally immunized in gnotobiotic rats and to compare the effect of two different delivery systems. METHODS: Sprague Dawley rats, infected with S. mutans at 20 days of age, were intranasally immunized with plasmid pCIA-P (group A), Dosper-DNA complex (group B), Bupivacaine-DNA complex (group C). Control rats were either immunized with plasmid pCI (group D), distilled water (group E) or immunized intramuscularly (group F). All the rats were boosted 2 weeks later. ELISA determined the antibodies against the vaccines. Keyes caries score was used to evaluate the anti- caries effectiveness of the vaccines at the terminal study. RESULTS: As for the antibody reactions, there were significantly (P < 0.01) differences between rats immunized with DNA vaccine and non-immunized rats. And rats in group B and C had the significantly (P < 0.01) higher level of specific salivary anti-PAc IgA antibodies and rats (group B, C, F) had the significantly (P < 0.01) higher specific serum anti-PAc IgG responses to DNA vaccine. Keyes scores of rats (group B and C) were significantly (P < 0.01) lower than others. CONCLUSIONS: Intranasal immunization with plasmid pCIA-P encoding pac gene successfully reduces the caries and appears to be a promising approach against dental caries. Cationic liposome Dosper and local anesthetic bupivacaine could enhance the efficacy of DNA vaccine. PMID- 12641963 TI - [Construction and cellular expression of GTF-PAc fusion anti-caries DNA vaccine]. AB - OBJECTIVE: To construct a fusion anti-caries DNA vaccine pGLUA-P carrying GLU fragment from gtfB gene of Streptococcus mutans GS-5 and A-P fragment including the A region and P region of PAc protein from a DNA anti-caries vaccine pCIA-P, and to investigate its expression in prokaryotic and eukaryotic cells. METHODS: The sequence of GLU fragment in pGLU plasmid was testified by DNA sequencing. The fusion anti-caries DNA vaccine was constructed by ligating A-P fragment from pCIA P to pGLU. The expression of GLUA-P fusion protein in E. coli BL21 (DE3) was induced by IPTG and checked by SDS-PAGE electrophoresis. pGLUA-P was transfected in vitro to cultured rat primary muscle cells by cation liposome Dosper, and immunohistochemical method was used to test the expression of GLUA-P fusion protein in cells. RESULTS: GLU sequence was identical with relative sequence of GTF-I (GS-5 strain) in Gene Bank. Recombinant eukaryotic expression plasmid pGLUA P was confirmed to have both GLU and A-P fragment. After pGLUA-P was transferred into E. coli (DE3), it could express a new 115 000 protein by the induce of IPTG. Specific brown products could be found in the cytoplasm of cultured rat primary muscle cells transfected by pGLUA-P. CONCLUSIONS: Fusion anti-caries DNA vaccine pGLUA-P is successfully constructed and confirmed by sequencing and enzymes digestion. Fusion GLUA-P protein can be correctly expressed in prokaryotic and eukaryotic cells. PMID- 12641964 TI - [Immunogenicity of a synthetic peptide vaccine from glucosyltransferase]. AB - OBJECTIVE: To measure the immunogenicity of a synthetic peptide of glucosyltransferase (GTF) for designing synthetic peptide-based vaccine of dental caries. METHODS: A fusion 27-mer peptide, containing the conserved regions within catalytic and glucan-binding domains of GTF, was synthesized. Serum antibodies to the synthetic peptide were determined by ELISA method. Inhibitions of both GTF activity and S. mutans adherence were detected for the functions of antisera. RESULTS: The sequence of fusion peptide vaccine was ANDVDNSNPVVQAEQLYFRANGVQVKG. The spleen weights of immunized mice were heavier than the control ones. Specific antibodies were effectually elicited. The immune sera not only inhibited GTF enzymatic activity but also inhibited the vitro adherence of S. mutans. CONCLUSIONS: The peptide vaccine which involves antibody-mediated inhibition of the catalytic and the glucan-binding activities of GTF may be valuable for controlling the dental caries. PMID- 12641965 TI - [A clinical evaluation of filling radiation caries]. AB - OBJECTIVE: To evaluate the clinical effects of filling for radiation carious cavities. METHODS: 146 carious cavities of 15 patients after radiotherapy for head and neck malignancies were included and were filled with two new GIC (Ketac Molar and Fuji IX). The 2 years follow-up results were evaluated with USPHS evaluation criterion. RESULTS: (1) The 2 years excellent rates were 51.7% in Ketac-Molar and 52.3% in Fuji IX, no significant differences. (2) Failures and defects were mainly occurred after 1 and 2 years. The main reason of failure was loss of filling. Marginal staining was a common shortcoming of the two materials. (3) There were no significant differences in main indexes except retention. CONCLUSIONS: It is successful for radiation caries to be filled with Ketac-molar and Fuji IX, and is practically significant in stomatological clinic. PMID- 12641966 TI - [Development of a computerized automatic identification system for use in cephalometry]. AB - OBJECTIVE: To establish the automatic X-ray cephalometric analysis system to simplify cephalometric steps and to provide a convenient and reliable method for cephalometric analysis. METHODS: The system which was programmed by visual-c language, and graphics and image processing techniques and artificial intelligence were used. The techniques related to computer digital image processing and pattern recognition such as Median filtering, Histogram equalization, Laplacian and Canny edge detection were introduced. It could automatically outline the contour lines of the hard and soft tissues by establishing the templates of the variable anatomical structures. RESULTS: The following functions were established: (1) automatically outlining the contour lines of the soft tissues. (2) automatically recognizing, measuring and analysing the landmarks of soft tissues. (3) automatically recognizing porion, sella and the landmarks of the mandible. (4) automatically building the contour lines of the hard tissues. In brief, the system used the more advanced methods, calculated more precisely and saved more time and energy than other systems. CONCLUSION: The system is a more convenient and precise tool in cephalometry. PMID- 12641967 TI - [A study on the stability and reliability of 3D laser cranio-maxillofacial reconstruction]. AB - OBJECTIVE: To study the stability and the reliability of the first 3D laser cranio-maxillofacial reconstruction system in China. METHODS: The same 3D laser scan image of a head plaster model was measured twice three weeks before and after. That head cast was also scanned and reconstructed twice three weeks before and after, and then the 3D images were measured. To the same head cast we compared the laser measurement with the direct manual measurement. The statistical analysis of measurements was made using the paired t test. RESULTS: There was no significant difference between two sets of measurements on the same image (P > 0.05). The difference in the measurements was not significant between two reconstructed images of the same model (P > 0.05). And there was also no significant difference between the measurements by laser and by hand (P > 0.05). CONCLUSIONS: The recognition of the author to the 3D laser scan image is reliable. The laser scanning system has a good stability and reliability and can be used in clinical study preliminary. PMID- 12641968 TI - [The detection of DNA adduct in human peripheral lymphocytes by bioluminescence method]. AB - OBJECTIVE: To develop bioluminescence method to measure DNA adducts in human peripheral lymphocytes and study the factors that affect the level of DNA adduct, in order to use it in studying the relationship between water pollution and human DNA adduct levels. METHOD: The measurement of DNA adduct in peripheral lymphocytes was carried out in 234 normal persons, and the relationship between DNA adducts levels and age, smoking and eating habit etc. was analyzed. RESULTS: (1) The level of DNA adduct in males was higher than that in females, but without significance (P > 0.05); (2) The level of DNA adduct was significantly correlated with age, smoking, tea drinking, eating habit and alcohol drinking. Among them, age plus smoking were the most important factors affected the DNA adduct levels (P < 0.01). The level of DNA adduct increased with age. The more cigarette smoked, the higher was the level of DNA adduct. CONCLUSION: The level of DNA adduct is affected by many factors, so it needs further research to apply it in the biomonitoring and risk assessment of environment chemicals. PMID- 12641969 TI - [Immunogenicity and safety of recombinant yeast-derived hepatitis B vaccine in adults]. AB - OBJECTIVE: To study the immunogenicity and safety of recombinant yeast-derived hepatitis B vaccine (YDV) in adults. METHODS: One hundred and twenty-four healthy teachers aged 22 approximately 58 years with serum negative HBsAg, anti-HBs, anti HBc and with normal temperature were randomly selected from Beipiao city, Liaoning province. All the subjects were immunized with 5 microg/0.5 ml of YDV made by Beijing Institute of Biologic Products, for three doses at an interval of one and six months, respectively. RESULTS: The positivity of serum anti-HBs was 35.0%, 83.3%, 65.5% and 32.7% with a geometric mean titre (GMT) of 12.6 mIU/ml, 402.0 mIU/ml, 70.3 mIU/ml and 20.3 mIU/ml, respectively, three, seven, 12 and 24 months after immunization. The positivity and GMT of serum anti-HBs appeared the highest seven months after immunization, then began to decrease sharply. The positivity and GMT of serum anti-HBs in women was higher than that in men either three, or seven, or 12, or 24 months after immunization. The positivity of serum anti-HBs in those of 35 years or over was lower than that less than 35 years, seven months after immunization, but no age difference could be found 12 months after immunization. No local or systematic adverse reactions were found in all the subjects within three days after immunization. CONCLUSION: The recombinant yeast-derived hepatitis B vaccine (YDV) is immunogenic and safe for adults, but the persistency of serum anti-HBs in after immunization should be followed-up further. PMID- 12641970 TI - [Inhalation of sulfur dioxide on the DNA damage of brain cells in mice]. AB - OBJECTIVE: To investigate the inhalation of sulfur dioxide (SO(2)) on the DNA damage of brain cells in mammalian animals. METHOD: Single cell microgel electrophoresis technique (comet test) was used to test the DNA damage of brain cells. RESULTS: (1) After the exposure to SO(2) at 0, 7, 14, 28 mg/m(3), the tail lengths of nuclear DNA in brain cells from male mice were 8.02, 23.14, 46.43 and 53.49 micro m respectively; and that from female mice were 7.23, 12.43, 20.39 and 54.83 micro m respectively. The results showed that: (1) SO(2) inhalation caused damage on DNA of brain cells in a dose-dependent manner; (2) Even under lower concentration of SO(2) as 7 mg SO(2)/m(3), the damage on DNA of brain cells was also reached to 98.8%. It implied that the brain cells of mammalian animals were very sensitive to SO(2) inhalation; (3) The DNA damage of brain cells from male mice is more serious than that from female mice. The reasons remain to be further studied. CONCLUSIONS: SO(2) pollution even at lower concentrations also had a potential risk to the genetic material DNA of brain cells from mammalian animals. The results of our study might explain the recently published epidemiological studies that the workers exposed to SO(2) or SO(2) derivatives had suffered an increase of mortality from brain cancer. PMID- 12641971 TI - [The analysis of medical-related cost for in-patients with injuries in Ningxia region]. AB - OBJECTIVE: To understand socio-economic losses of inpatients and deaths caused by injuries in 2000 in Ningxia and to estimate their extent of harmfulness. METHODS: Eight of 35 local hospitals totaling 5 876 inpatients were recruited with two stage sampling in Ningxia in 2000. All medical cost incurred during their hospitalizations for injuries, cardiovascular and cerebrovascular diseases, respiratory disease, cancer and communicable diseases, losses in labor time were analyzed, and years of potential life lost (YPLL), working years of potential lost (WYPLL), valued years of potential life lost (VYPLL) due to these diseases were estimated for the residents in Ningxia with corrected human capital method. RESULTS: The study showed that indirect economic losses due to hospitalization for injuries accounted for 24 million yuan, higher than those for other diseases. YPLL, WYPLL and VYPLL due to injuries were also higher than those in other diseases. CONCLUSIONS: Injury has caused serious threat to their health of the residents in Ningxia and brought heavy burden for the society and economy. It has become an important public health problem and its prevention and control should be strengthened as soon as possible. PMID- 12641972 TI - [Risk factors in women with gestational diabetes mellitus]. AB - OBJECTIVE: To identify risk factors for gestational diabetes mellitus (GDM) in women and to study the contribution of family history of type-2 diabetes to the risk for DDM. METHODS: A case-control study was performed in 85 women with GDM and 177 cases controls. Univariate and multivariate logistic regression and log linear model were used to identify risk factors of GDM. RESULTS: Multivariate logistic regression showed that obesity before pregnancy, family history of type 2 diabetes, birth weight of pregnant women, age, fasting plasma level of triglyceride, physical inactivity, etc. all were risk factors for GDM. Analysis with log-linear model showed that parents' (father's or mother's) history of type 2 diabetes associated with GDM, with P-values of 0.012 and 0.017, respectively. Prevalence of diabetes in the mothers of proband with GDM was 9.41%, as compared with that in the fathers of proband with GDM 8.24%, with no statistical significance. CONCLUSIONS: Obesity before gestation, family history of type-2 diabetes, low birth weight of mother, age, increased fasting plasma level of triglyceride, as well as parents' history of type-2 diabetes, all were risk factors for GDM. Physical exercise was found to be a protective factor for GDM. Mother's history of type-2 diabetes did not differ from father's in contributing to the onset of GDM in their offspring. PMID- 12641973 TI - [Application of reverse transcriptase polymerase chain reaction for surveillance of influenza virus in Beijing]. AB - OBJECTIVE: To establish the RT-PCR method to type the influenza virus. METHODS: After amplifying the virus by cell culture, we carried out RT-PCR by using two pairs typing primers and four pairs subtyping primers to detect the influenza virus. RESULTS: In those 23 samples which had cytopathologic changes, there were 10 positive strains detected by RT-PCR assay including seven A type (six H3N2 subtype and one H1N1 subtype) and three B type. CONCLUSION: This method is rapid, specific and sensitive and possesses great value for practical application in the surveillance of influenza virus. PMID- 12641974 TI - [Exposure level of total N-nitroso compounds in residents of high-and low-risk areas for esophageal cancer in southern]. AB - OBJECTIVE: To assess the exposure level of total N-nitroso compounds (TNOCs) in the residents of high- and low-risk areas for esophageal cancer in southern China. METHODS: Samples of duplicate plate diets and 12 hr overnight urine were collected from 120 male adults in each of the 2 areas, a high-risk area (Nanao county) and a low-risk area (Lufeng county) for esophageal cancer. The 240 male healthy subjects (35 - 64 years old) were selected by a 3-stage random cluster sampling procedure. Levels of TNOC, N-nitrosamino acids (NAAs) and volatile N nitroso compounds (VNOC) in the samples were measured by Thermo Energy Analyzer. RESULTS: The detectable rate (95%) of diet TNOC, daily dietary TNOC intake (4.25 +/- 0.84) micromol/day, 12-hr urinary TNOC excretion levels (1.76 +/- 0.23 ng/12 h) and daily dietary intake of VNOC (266 +/- 31.2 microg/day) in the high-risk area were all significantly higher than those of the low-risk area. Oesophageal cancer mortality rates were positively and significantly associated with daily dietary TNOC intake and 12-hr urinary TNOC excretion. Urinary NAAs excretion levels were not different in the two areas. CONCLUSION: The results suggest that TNOCs may be implicated in the etiology of esophageal cancer in southern China. PMID- 12641975 TI - [Effect of neuroprotectant agent combined with cocktail on expression of anti apoptotic protein bcl-2 in rats after focal cerebral ischemia]. AB - OBJECTIVE: To investigate whether the protective effect of therapy with different combined neuroprotectant agents was better than that of single agent on focal cerebral ischemia. METHODS: The right middle cerebral artery in the rats was occluded with suture occlusion technique. The rats were divided into five groups treated with FDP (50 mg/kg, n = 10), MK-801 (1 mg/kg, n = 10) and NAC (150 mg/kg, n = 10) singly, or in combination, respectively, by intraperitoneal infusion 30 minutes after vessel occlusion. The rats were weighed and assessed neurologically, based on a 5-point scale, six and 24 hours after focal cerebral ischemia. The expression of anti-apoptotic protein bcl-2 was observed with SDS PAGE protein electrophoresis and Western blot technique. RESULT: The optical density of bcl-2 increased more distinctly in the rats treated with combined neuroprotective agents than that with any single agent six and 24 hours after cerebral ischemia, with a statistically significant difference (P < 0.05). CONCLUSIONS: Treatment with combined neuroprotectant agents could un-regulate the anti-apoptotic protein bcl-2 more distinctly than that with any single agents. Combined use of neuroprotectants might be more effective than that of single agent in protecting rats' brain from ischemia. PMID- 12641976 TI - [The effects of exposure to rare earth (NO3)3 on the immune function of mice off spring via milk]. AB - OBJECTIVE: To study the changes of immune function in mice offspring whose mothers were exposed to Rare Earth (RE)(NO(3))(3). METHODS: RE(NO(3))(3) was administered to mother mice after giving birth by gavage at dosages of 2, 20 and 200 mg/kg bw during breast-feeding period. The weights of spleen and thymus, the spleen plaque forming cells (PFC), the delayed type hypersensitivity (DTH) and the charcoal clearance of the offspring were determined. RESULTS: The results obtained from the offspring after weaning showed that the body weight of offspring treated with 200 mg/kg RE(NO(3))(3) was 18.8% lower than that of the control group; at the dosage of 2 mg/kg, the number of IgM-PFC was increased by 82.7%; and at the dosage of 20 mg/kg the rate of clearance and clearance index were significantly higher than that of the control group. No difference in DTH was found in any treated group as compared to the control group. The results of offspring at three weeks after weaning showed that the number of IgM-PFC of the 20 and 200 mg/kg bw dose groups were 47.0% and 44.7% lower than that of control group respectively; the rate of clearance and clearance index of the 200 mg/kg group were significantly lower than that of the control group. No significant changes in DTH were observed in each exposed group. CONCLUSION: RE(NO(3))(3) treatment affected the immune function of mice offspring which may caused by breast milk. PMID- 12641977 TI - [Effect of 7,12-dimethylbenz(a)athrancene on immune function in metallothionein gene-knocked-out mice]. AB - OBJECTIVE: To study the immunotoxicity induced by 9,10-dimethyl-1,2 benzathrancene (DMBA) in metallothionein gene-knocked-out mice [MT(-/-)] as compared with that in wild-type mice [(MT(+/+)]. METHODS: Female mice were treated with 25 mg/kg and 50 mg/kg of DMBA i.p., respectively and immunized with sheep red blood cells (SRBC) i.v. on the following day and rechallenged by injection of SRBC via footpad s.c. on the fourth day post-immunization. Humoral and cell-mediated immune function was assessed by the number of spleen IgM antibody plaque formation cells (PFC) to SRBC and cell-mediated delayed-type hypersensitivity (DTH) measured by footpad swelling thickness. RESULTS: After treatment with 25 mg/kg DMBA, a decrease in weight of their spleen and thymus and PFC/spleen were observed in MT(-/-) mice, while only decrease in thymus weight of MT(+/+) mice. The humoral function was suppressed by 72% in MT(-/-) mice. No obvious change in cell-mediated immune function was observed both in MT(-/-) and MT(+/+) mice. Both humoral and cell-mediated immune function were suppressed more severe (91%) in MT(-/-) mice treated with 50 mg/kg DMBA than those treated with 25 mg/kg DMBA (72%). DTH was not altered by DMBA in MT(+/+) mice. The weight of their spleen and thymus decreased and humoral immune function suppressed in MT(+/+) mice, but these changes were significantly less severe. No obvious suppression of cell-mediated immune function was observed in MT(+/+) mice. CONCLUSION: Their humoral and cell-mediated immune function was more susceptible to being suppressed by DMBA in MT(-/-) mice, indicating that MT could protect their immune function from damage caused by DMBA. PMID- 12641978 TI - [Dynamic monitoring of neural tube defects in China during 1996 to 2000]. AB - OBJECTIVE: The database from Chinese Birth Defects Monitoring Network was used to describe the epidemiological features and secular trends of neural tube defects (NTDs) prevalence during Jan. 1996 to Dec. 2000, including anencephaly, spina bifida and encephalocele. METHODS: Data were collected with hospital-based cluster sampling method. During the period, all live or still births with 28 weeks or more of gestation were assessed within seven days after delivery. RESULTS: There were 2 873 case with NTDs identified from 2 281 616 births, with an overall prevalence rate of 12.95 per 10,000 births. And, the prevalence rates of anencephaly, spina bifida and encephalocele were 5.02 per 10,000, 6.30 per 10,000 and 1.64 per 10,000, respectively. Significant falls in overall prevalence rates of NTDs and of anencephaly were observed. The prevalence rates of NTDs were 9.75 per 10,000 and 15.96 per 10,000 in male and female births, respectively, 7.76 per 10,000 and 25.20 per 10,000 in the urban and rural areas, respectively, and 19.90 per 10,000 and 5.81 per 10,000 in north and south China, respectively. The prevalence was higher in the groups with maternal age of less than 20 years and more than 30 years than in those other maternal ages. Preterm babies and babies with low birth weight accounted for 50.9% and 50.6% of perinatal babies with NTDs, respectively, with a perinatal mortality of 77.8%. CONCLUSIONS: In China, the occurrence of NTDs was higher in female births than in male births, higher in the rural than in the urban, and higher in the north than in the south. Annual prevalence rate of NTDs in China presented a declining trend, but still higher than that in the other countries at the same time period. Babies with NTDs were in poor birth quality, higher perinatal mortality and poor prognosis. Effective preventive measures and prenatal diagnosis should be strengthened to reduce the occurrences of neural tube defects. PMID- 12641979 TI - [Effects of chrysotile asbestos on the activities of cytochrome P4501A1 and glutathione S-transferase in A549 cell line]. AB - OBJECTIVE: To study the effects of chrysotile on the activities of some enzymes for xenobiotics metabolism. METHODS: UICC chrysotile (UC) and China Mangya chrysotile (MC) asbestos fibers were used at different doses (0, 25, 50, 100, 200 mg/L) to study its effects on the activities of cytochrome P4501A1 and glutathione S-transferase (GST) in A549 cell line. Also, the effects of chrysotile on the activities of CYP1A1 and GST induced by benzo(a)pyrene were studied. RESULTS: The activity of EROD increased slowly with the increasing dose of UC, as A549 cells were incubated with UC for 24 h, and EROD activity increased by 40% at a dose of 200 mg/L UC. However, activity of EROD decreased by 32% with 48 h incubation at the same dose, indicating that lower dose of UC and short time could induce the activity of EROD in A549 cells, whereas higher doses and long time could inhibit its activity. MC exhibited a multiphasic effects on the activity of EROD, whether at a dose of 25 mg/L for 24 h or 48 h all gave the strongest induction, 1.86 times or 1.28 times as controls, respectively. However, EROD activity decreased with the increases in MC doses and incubation time, with the lowest as 35% of the controls. The effect of UC on GST activity was not so obvious, with the highest as the increase in its activity by 20%. The highest induction of MC to GST activity was at a dose of 25 mg/L, 2, 5 times as that in controls. With the increases in its doses, effects of MC on GTS activity became inhibition from induction, like that on EROD activity. MC at a dose of 200 mg/L could lower the activity of GST by 18.7%. A549 cells were incubated with chrysotile fiber for 24 h firstly, and then incubated with benzo(a)pyrene to induce the activities of EROD and GST. The results showed that neither UC nor MC could affect the activity of EROD induced by benzo(a)pyrene. However, UC at a dose of 200 mg/L and MC at 100 mg/L could increase the activity of GST induced by benzo(a)pyrene. CONCLUSION: Chrysotile at different doses or its different types showed varied effects on the activities of EROD and GST in A549 cell lines, probably because of different physicochemical characteristics and surface activity of two kinds of asbestos. PMID- 12641980 TI - [In situ re-grafting of the scalp skin flap after its covering of the exposed devitalized skull following electrical injury]. AB - OBJECTIVE: To investigate the clinical therapeutic effects of in situ re-grafting of the scalp skin flap after its covering of the exposed devitalized skull following electrical injury. METHODS: The scalp wounds were debrided during the early postburn stage and the necrotic skull was preserved. The wounds with necrotic skull were then covered with an adjacent scalp skin flap. The grafted scalp skin flap was re-grafted back to the donor site 3 - 6 months after the first operation. The remaining scalp wound with fresh granulation tissue was recovered with split-thickness skin grafts. RESULTS: Ten scalp skin flaps were applied in 8 cases of electrical injury of the skull with the maximal defect of 24 cm x 10 cm and all survived very well free from infection or necrosis. The wounds were closed primarily. CONCLUSION: In situ re-grafting of the scalp skin flap after its covering of the exposed devitalized skull following electrical injury shortened the wound healing time with satisfactory contour. PMID- 12641981 TI - [The repair of bulky tissue defect of forearm with skin flaps]. AB - OBJECTIVE: To evaluate the repairing methods of bulky tissue defect of forearm with flaps. METHODS: Twenty-one burned patients with wounds in the forearm were enrolled in this study. The injury causes were high-voltage electricity, hot press or crush injury. After local debridement, the forearm defects were repaired with pedicled complex flaps, latissimus dorsi musculocutaneous island flaps or large thoraco-abdominal flaps immediately. RESULTS: All the flaps survived very well with satisfactory results except for 1 patient in whom local ischemic necrosis and sub-flap infection at the distal end of the flap occurred. CONCLUSION: Early debridement followed by skin flaps with pedicles or musculocutaneous flaps transfer could be simple, safe and reliable treatment strategies in the management of bulky tissue defects of the forearm due to burn, electric injury, or other devastating injuries. PMID- 12641982 TI - [The applied anatomy of axial subdermal vascular network skin flap and its application in the management of deep burn wound]. AB - OBJECTIVE: To explore the anatomical basis and clinical application of axial subdermal vascular network skin flap (ASVNF) in the management of deep burn wounds. METHODS: Twelve samples of skin and subcutaneous tissue of lower extremities from adult cadavers were anatomically observed. The course, branching and the architecture of the subdermal vascular network (SVN) in the superficial facia and the ultra-thin area of the anterolateral femoral and saphenous axial subdermal vascular network skin flaps were intensively studied. The wounds in twenty-six patients with deep burn injury in the limbs were repaired with ASVNF. RESULTS: The circular thinning method was employed in the preparation of anterolateral femoral ASVNF whose blood supply was provided by the musculocutaneous perforators. And the "U" shaped ultra-thin method was employed in the preparation of saphenous ASVNF, whose blood supply was directly provided by skin vessels. The flaps in these 26 patients survived very well in terms of the flap color, texture, contour and function after followed-up for 0.5 approximately 2.5 year. CONCLUSION: Thinning methods for different kinds of ASVNF should be individualized. ASVNF might be an optimal choice in the management of deep burn wounds due to its obvious advantages. PMID- 12641983 TI - [Effect of different nutritional routes on the intestinal mucus barrier in scalded rats]. AB - OBJECTIVE: To observe the effect of different nutritional routes of giving nutrition on the intestinal mucus barrier in severely scalded rats. METHODS: Wistar rats inflicted with 30% TBSA III degree scalding on the back were employed as the model and were randomly divided into 3 groups, i.e. control (C), parenteral nutrition (PN) and enteral nutrition (EN) groups. The rats in PN and EN groups were supplied with equal amount of nitrogen and calories and with equal volume of nutrition solution. The dynamic changes in the thickness of intestinal mucus layer and the contents of protein, hexose and acetylneuraminate in the mucus were examined. RESULTS: When compared with those in C group, the intestinal mucus layer became thinner and the contents of protein, hexose and acetylneuraminate in the mucus in both PN and EN groups decreased evidently after scalding. When compared between two nutritional groups, the thickness of intestinal mucus layer and the contents of the hexose and acetylneuraminate in the mucus in EN were much thicker and higher than those in PN group, while the mucus protein content exhibited no obvious difference between PN and EN groups. CONCLUSION: It was suggested that intestinal goblet cell synthesized and secreted less mucus after scalding in rats resulting in thinning of intestinal mucus layer and the change in mucus components. When compared with those in PN group, less injury to the intestinal goblet cells occurred and the intestinal mucus synthesis was less affected in EN group, and the components of intestinal mucus were maintained stable. PMID- 12641984 TI - [The effects of glutamine dipeptide on the improvement of endotoxemia in severely burned patients]. AB - OBJECTIVE: To explore the influence of glutamine dipeptide on the plasma endotoxin levels in severely burned patients. METHODS: Thirty burned patients with TBSA of 30 - 70% and III degree burn area more than 20% were randomly divided into control (C) and study (S) groups. Glutamine dipeptide powder in dose of 0.5 g/kg/day was given orally in bolus to those patients in S group during 1 - 12 postburn days (PBDs). The plasma levels of glutamine were determined during 1 12 PBDs. Simultaneously, the plasma endotoxin level was detected on 1, 3, 6 and 12 PBDs. The wound healing rate at 30 PBD and total hospital stay days were recorded. RESULTS: The plasma glutamine levels at 1 PBD in C and S groups were obviously lower than normal level (659.5 +/- 35 micromol/L), but there was no difference between these two groups (P > 0.05). The plasma glutamine levels in C group was much lower than that in S group at 12 PBD (P < 0.05). The plasma endotoxin concentration on 1 PBD in these two groups increased evidently compared with the normal value (P < 0.05), and there was no difference between the two groups (P > 0.05). The plasma endotoxin level in S group was much lower than that in C group on 3 PBD (P < 0.05). As for the wound healing rate at 30 PBD, it was markedly higher in S group than that in C group (91% vs 85%). On the other hand, the hospital stay days in S group were evidently lower than that in C group (52 vs 67). CONCLUSION: Oral intake of glutamine dipeptide in burn patients could be beneficial to the maintenance of the plasma concentration of glutamine and in decreasing plasma endotoxin level. It would also enhance the wound healing rate at 30 PBD and shorten the hospital stay days. PMID- 12641985 TI - [The influence of high frequency partial liquid ventilation on the cardiopulmonary function in dogs with inhalation injury]. AB - OBJECTIVE: To investigate the influence of high frequency partial liquid ventilation (HFJV) on the cardiopulmonary function in dogs with inhalation injury. METHODS: Sixteen mongrel dogs inflicted by hot steam inhalation were subjected to severe inhalation injury and were randomly divided into control (C) and treatment (T) groups. The dogs in both groups were all given HFJV. In addition, the dogs in T group were simultaneously supplied with perfluorocarbon liquid (3 ml/kg) into the lungs slowly via tracheal intubation for liquid ventilation. The blood gas analysis, pulmonary compliance, airway resistance and hemodynamic parameters were determined at 30, 60 and 90 minutes after ventilation. RESULTS: The PaO(2) in T group increased progressively, which was significantly higher than the post-injury value at all time points (P < 0.05). While the PaO(2) in C group exhibited no difference to the post-injury value at all time points. The PaCO(2) in T group increased obviously and was higher than the post-injury value at 60 and 90 post-ventilation minutes (P < 0.05). Furthermore, the PaO(2) in all the time points in T group was a little higher than that in C group (P > 0.05) and PaCO(2) in T group was much higher than that in C group at 90 min after ventilation (P < 0.05). But there was no difference between the two groups in terms of dynamic/static pulmonary compliance and airway resistance as well as the hemodynamics. CONCLUSION: Compared with simple HFJV, high frequency partial liquid ventilation seemed to be beneficial to the oxygenation after inhalation injury and to be no influence on the hemodynamics. PMID- 12641986 TI - [An experimental study on the role of early postburn massive escharectomy on the prevention of internal organ dysfunction]. AB - OBJECTIVE: To explore the role of massive escharectomy at early postburn stage in the prevention of internal organ dysfunction. METHODS: (1) Ten cases of severely burned patients were randomly divided into early (A) and non-early escharectomy (B) groups in equal number. Venous blood samples were harvested from the patients of the two groups in 1, 3 and 7 postburn days (PBDs), And the samples from 6 healthy volunteers were taken as the control. The serum was separated from the above blood samples and was employed to stimulate cultured HUVECs in vitro. The cell viability and permeability was observed after the stimulation. (2) Seventy Wistar rats inflicted with 30% TBSA III degree scalding were used as an animal model, and were randomized into early (C, n = 30) and non-early escharectomy (D, n = 30) groups, with 5 normal rats as control in each group. Intra-peritoneal fluid infusion was carried out at 1, 3, 6, 12, 24 and 48 postburn hours (PBHs) in rats in both groups. The rats were killed by blood letting at 1 hour after fluid supplementation. The changes in peritoneal macrophage (M Phi) activation state and plasma contents of LPS, IL-8, PLA(2) and MDA were determined at 48 hours after escharectomy in the rats. RESULTS: The cell viability and permeability of the HUVECs co-cultured with the serum from burn patients in E group was much better preserved than that in B group. On the other hand, the peritoneal M Phi activation and the plasma contents of LPS, IL-8, PLA(2) and MDA in C group were obviously decreased compared with those in D group. CONCLUSION: Early postburn escharectomy to remove denatured burned tissue were proved to be helpful in ameliorating endothelial injury and in inhibiting activation of inflammatory cells. Therefore, early escharectomy was assumed to be beneficial in the prevention of postburn SIRS and MODS. PMID- 12641987 TI - [Comparison of three regression methods in the calculation of LA(50)]. AB - OBJECTIVE: To search for an optimal regression method for LA(50) in burn management. METHODS: Three kinds of regression methods, i.e. linear regression, probability type probit regression and logic type probit regression. were employed to calculate the LA(50) and its 95% confidence interval (CI) from different burn data. The results produced by these three methods were compared with one another. RESULTS: Three methods were used to calculate LA(50) of one sample with the same, different or evidently different LA(50), but the order of LA(50) from high to low values appeared to be the same (P > 0.05). When the 95% CI of LA(50) was calculated by three methods, significantly different results were obtained (P < 0.01), especially the 95% CI by linear regression was evidently different to that produced by two other kinds of probit regression (P < 0.01), while there was no difference in the 95% CI between the two probit regression methods (P > 0.05). CONCLUSION: Different methods for the calculation of LA(50) and 95% CI came to different results. The LA(50) from different samples could only be compared as valid when the same calculation method was employed. Probit regression (logic type) seemed to be the best method in the calculation of LA(50) in terms of its simplicity, practicability and accuracy. PMID- 12641988 TI - [The role of C5a in adhesion properties of polymorphonuclear leukocyte to pulmonary vascular endothelial cells in burn patients with acute lung injury]. AB - OBJECTIVE: To explore the postburn adhesion properties of polymorphonuclear leukocyte (PMN) onto pulmonary vascular endothelial cells (PVEC) in burn patients with acute lung injury (ALI), so as to determine the role of C5a on PVEC-PMN adhesion. METHODS: Microtubule sucking technique was employed to determine the PVEC-PMN adhesion. The myeloperoxidase (MPO) was also assayed to reflect the magnitude of PVEC-PMN adhesion. RESULTS: The magnitude of PVEC-PMN adhesion increased and the adhesion force increased along with an increase in rh-C5a concentration. Simultaneously, the MPO activity was increased, which could be inhibited by anti-C5aR McAb in a concentration 1:104. CONCLUSION: Both C5a and C5aR participated in PVEC-PMN adhesion, which might be important in the pathogenesis of ALI. PMID- 12641989 TI - [An observation of the basement membrane remodeling after the combined grafting of xenogenic acellular dermal matrix with autoskin in rats]. AB - OBJECTIVE: To observe the dynamic process of basement membrane remodeling after the combined grafting of xenogenic acellular dermal matrix with autoskin. METHODS: The rat skin wounds were covered with xenogenic porcine acellular dermal matrix overlaid with razor thin autoskin. The skin samples were collected at 1, 2, 3, 4, 8, 12 and 16 post-grafting weeks. The changes in laminin expression in the basement membrane and the ultrastructure of the basement membrane at 12 post grafting weeks were observed by immunohistochemistry and transmission electron microscopy. The results were compared with those in simple thin autoskin grafting as the control. RESULTS: The laminin expression in the combined grafting was higher than that in control. At 12 post-grafting weeks, the basement membrane in combined grafting rats was clear and continuous and the hemidesmosome was relatively more in amount and distributed evenly. While in the autoskin group, the lamina densa in the basement membrane was blurred and discontinuous with a decrease in and uneven distribution of hemidesmosome. CONCLUSION: The increased expression of laminin in the basement membrane in the combined grafting rats might be beneficial to the remodeling of the basement membrane and to strengthening the connection of epithelium to the dermis, thus wound healing quality would be improved. PMID- 12641990 TI - [The relationship between intestinal bifidobacteria and bacteria/endotoxin translocation in scalded rats]. AB - OBJECTIVE: To investigate the potential role of intestinal bifidobacteria in the pathogenesis of gut-origin bacteria/endotoxin translocation in scalded rats. METHODS: Wistar rats inflicted with 30% III degree scalding on the back were employed as the model with the rats undergoing sham injury as the control. The intestinal bacteria/endotoxin translocation and the changes in cecal mucosal microflora were determined by routine methods. And the plasma IL-6 concentration was measured with ELISA. RESULTS: The incident of bacterial translocation into internal organs increased markedly in scalded rats (P = 0.001). The plasma LPS levels on 1, 3 and 5 postburn days (PBDs) in scalded rat group were much higher than those in sham injury group. The number of bifidobacteria decreased sharply 20 - 250 fold, the fungi increased 5 - 60 fold and E. coli increased 0.5 - 30 fold in the caecal mucosal microflora in the scalding group. The ratio of bifidobacteria to E. coli in the scalding group (4 - 800:1) was much lower than that in the sham injury group (25000:1). Furthermore, the plasma IL-6 level increased evidently in the scalding group. It was indicated by further analysis that compared with the rats without bacterial translocation, the bifidobacteria decreased 120 fold, the fungal number increased 50 fold and the E. coli number increased 30 fold in the scalded rats. The bifidobacterial number in the caecal mucosal microflora was negatively correlated with the plasma concentrations of IL 6 and LPS (P < 0.01) in the scalding rat group, and the plasma concentration of IL-6 was significantly and positively correlated with that of LPS. CONCLUSION: Severe scalding injury could lead to an the imbalance of intestinal microflora and the increased intestinal translocation of bacteria and LPS. The decrease of the ratio and number of bifidobacteria in the caecal mucosal microflora might be a contribute to the occurrence of postburn intestinal bacteria/endotoxin translocation. PMID- 12641991 TI - [Study on the relationship between epidermal stem cells and the developing process of sweat gland in human fetal skin]. AB - OBJECTIVE: To explore the relationship between epidermal stem cells and the developing process of sweat gland in human fetal skin, so as to obtain a hint for future induction of epidermal stem cells to differentiate into sweat gland cells. METHODS: Total layer of human skin from the back of fetus at gestational ages from 11 to 31 weeks, obtained from spontaneous abortion was routinely examined. The expressions of beta1 integrin and keratin 19 in sweat gland cords or buds and mature sweat gland cells were dynamically observed with SP immunohistochemical technique. The development and maturation of sweat gland were identified by the positive staining of keratin 8 with immunohistochemistry. RESULTS: It was revealed by histologic observation that basal layer cells of the primary epidermal ridge exhibited focal aggregation and formed hillocks at 16 gestational weeks. The hillocks of cells then migrated downward as cords into the dermis during 18 - 20 gestational weeks. Then, the end part of the cell cord developed into a round lump of twining cords assuming the mature sweat gland. The expressions of beta1 integrin and keratin 19 were found not only in sweat gland cords and buds but also in the mature cells and lasted throughout the total period of sweat gland development. The expression of keratin 8 in sweat gland buds started since 14 - 16 gestational weeks and maintained thereafter. CONCLUSION: The sweat gland started to develop during 14 - 16 gestational weeks and matured at 24 weeks. During the development process of sweat gland, epidermal stem cells were considered to be the key source. PMID- 12641992 TI - [An observation of repair of burn wound with consanguineous skin pretreated with Tripterygium wilfordii]. AB - OBJECTIVE: To explore new source of skin for burn wound coverage. METHODS: Split thickness consanguineous skin was harvested from New Zealand white rabbit and was soaked in 200 g/L of multi-peptides of Tripterygium wilfordii, 50 g/L of dexamethasonel, on 9 g/L of normal saline solution for 15 - 30 mins, respectively. The consanguineous skin was thereafter grafted onto the whole layer skin defects in filial generation of rabbits with non-consanguineous skin as the control. The survival time and rejection of the grafted skin was observed. RESULTS: The rejection appeared evidently less intense and survived significantly longer (43 +/- 3.5 days) when the consanguineous skin was pretreated by Tripterygium wilfordii. However the grafted consanguineous skin survived for 30 +/- 2.5 days when it was pretreated by dexamethasone. The grafted skin was quickly rejected and survived only for 11 +/- 1.6 days when the skin was pretreated by normal saline or the skin was non-consanguineous. CONCLUSION: Consanguineous skin possessed partial compatibility with the recipient due to similar antigen, which was beneficial to the its survival, especially after the skin was pretreated. PMID- 12641993 TI - Evidence-based Medicine in Infectious Diseases and Microbiology: Still a Long Way to Go. PMID- 12641994 TI - Gastrointestinal Infections Caused by Cytomegalovirus. AB - Cytomegalovirus (CMV) is an important human pathogen, causing a variety of syndromes ranging from asymptomatic infections to life-threatening end-organ disease. Infections of the gastrointestinal tract are an especially common form of disease caused by CMV and are usually manifest as luminal infections, such as esophagitis or colitis. Solid organ disease caused by CMV is also known to occur, with hepatitis being the most common syndrome. The large majority of cases of tissue-invasive disease caused by CMV have been reported to occur in immunocompromised hosts. Patients at highest risk for serious CMV-mediated disease are commonly HIV infected, recipients of transplants, or are receiving potent immunosuppressive medications. Intriguingly, several recent reports have discussed the previously underappreciated role of serious forms of CMV-mediated disease occurring in immunocompetent patients. This paper reviews several recent reports addressing important aspects related to the diagnosis, treatment, and management of CMV infections of the gastrointestinal tract. PMID- 12641995 TI - The Occurrence and Prevention of Infections Associated with Gastrointestinal Endoscopy. AB - In the four decades since the introduction of flexible endoscopy into medical practice, nearly 300 cases of human infections or pseudoinfections involving bacteria, fungi, parasites, and viruses have been linked to endoscopic procedures. In the majority of such cases, inadequate cleaning and disinfection techniques during the reprocessing of the instruments or their accessories have been likely contributing factors. Working groups from major gastroenterology societies and infection control organizations have established standards of care for the routine maintenance of endoscopic equipment in order to decrease the rates of infection even further. Since the institution of these standards, rates of transmission of infections to patients have decreased, though have not been completely resolved. This article reviews the available literature on transmission of pathogenic agents through endoscopic procedures, summarizes the current guidelines for the care of endoscopic equipment, and discusses available preventive measures aimed at decreasing the risk of endoscopy-related infections. PMID- 12641996 TI - Glutamine Analogues As Adjunctive Therapy for Infectious Diarrhea. AB - Glutamine is the major fuel for the gut as well as for many cells in the immune system that becomes conditionally essential during catabolic states. Glutamine supplementation improves intestinal mucosal repair and function. Glutamine, even at high doses, is without side effects and is well tolerated. Though unstable in solution, this is overcome by creating stable dipeptides such as alanyl glutamine. In HIV-positive patients with wasting, glutamine enhances intestinal absorptive function and weight gain. Glutamine enhances sodium and water absorption in a rabbit model of cholera and Cryptosporidium-infected piglet intestine. Both glutamine and alanyl-glutamine have recently proven effective in a bovine model of Cryptosporidium as well. Finally, a rat model of cholera toxin induced diarrhea also showed that alanyl-glutamine enhanced water and electrolyte intestinal absorption even better than the traditional glucose solutions. Clearly glutamine and its stabler derivatives hold promise for enhancing repair of mucosal injury by a wide range of infections or toxic agents, and hence have great potential as a new oral rehydration and nutrition therapy for patients with enteric infection, malnutrition, or chemotherapy- or radiation-induced enteritis. PMID- 12641998 TI - Glycoprotein D-adjuvant vaccine to prevent genital herpes. PMID- 12641997 TI - Current Concepts in the Management of Fungal Peritonitis. AB - Fungal pathogens are uncommon isolates in the setting of peritonitis. Secondary peritonitis results from a breach in the gastrointestinal tract with gross contamination of the peritoneum. Peritonitis in patients undergoing peritoneal dialysis represents a unique form of secondary peritonitis often caused by nosocomial bacteria and fungi. Regardless of the clinical circumstances, most cases of fungal peritonitis are caused by Candida species, however, other yeasts and filamentous fungi have been uncommonly reported. Treatment of secondary peritonitis consists of appropriate surgical intervention and systemic antifungal therapy. Systemic antifungals such as amphotericin B or fluconazole are also essential for the treatment of fungal peritonitis in patients who are peritoneal dialysis dependent. Salvage of the peritoneal dialysis catheter may be attempted, however, removal is usually required to achieve cure. Prophylaxis with fluconazole in patients with recurrent gastrointestinal perforations or anastomotic leakages has reduced the incidence of Candida peritonitis. Benefit of nystatin for fungal peritonitis prophylaxis in peritoneal dialysis patients is questionable. PMID- 12641999 TI - Trichomoniasis: What's New? AB - Trichomonas vaginalis is the most common nonviral sexually transmitted disease in the world. Recent research has elucidated several virulence factors that allow T. vaginalis to adhere to extracellular host proteins and evade the host immune system. Further investigation is needed to determine the mechanisms responsible for the perinatal morbidity associated with trichomoniasis and to evaluate the interactions between trichomoniasis and HIV infection. An effort to decrease T. vaginalis associated with perinatal morbidity by metronidazole treatment in pregnancy was unsuccessful. However, more sensitive tools for diagnosis such as polymerase chain reaction may further facilitate future understanding of the epidemiology and risk factors of disease and improve approaches to control this widespread infection. Treatment has remained highly efficacious with single-dose metronidazole, although concern regarding drug resistance has led to more systematic evaluations of the prevalence of metronidazole resistance and alternatives to metronidazole treatment. PMID- 12642000 TI - Sexually Transmitted Diseases in Men Who Have Sex with Men: A Clinical Review. AB - Increases in sexually transmitted diseases (STDs) among men who have sex with men (MSM) have coincided with recent increases in sexual risk behaviors across the United States and Europe. The identification of same-sex sexual risk behavior in men and the subsequent risk for certain bacterial and viral infections requires competency in taking a sexual history. Recent advances in the diagnosis and treatment of STDs have made STD management easier for physicians and patients and expanded the ability of a variety of health care professionals to participate in the management of STDs. This review focuses on recent developments in the epidemiology, pathogenesis, diagnosis, and management of common STDs in MSM. PMID- 12642001 TI - Sexually Transmitted Diseases in Men Who Have Sex with Men: An Epidemiologic Review. AB - The introduction of highly active antiretroviral therapy (HAART) has led to dramatic reductions in morbidity and mortality due to HIV infection. However, the resulting optimism and improved health status produced by HAART appears to have contributed to unanticipated consequences in men who have sex with men (MSM): loss of fear of acquiring and transmitting HIV, an increase in high-risk sex, decreased use of condoms, and a resurgence of gonorrhea and syphilis. Other factors, such as lack of knowledge of sexually transmitted diseases (STDs), use of the Internet as a venue to find sex partners, the increasing use of Viagra (Pfizer, New York, NY) as a recreational drug, and the apparent expanding role of oral sex in STD transmission are fueling these trends. Since ulcerative and inflammatory STDs facilitate HIV transmission, a new wave of HIV infection in MSM may be on the horizon. The rising STD rates and relapses in high-risk sexual behaviors in MSM, both HIV-infected and -uninfected MSM, have profound implications for public health and the clinical management of these patients. Clinicians should be aware of this turn of events, and implement new screening and counseling guidelines that have been issued in response to these alarming reports. PMID- 12642002 TI - Current Issues in Screening for Chlamydia trachomatis. AB - During the past several years, research related to screening for Chlamydia trachomatis has flourished, with new diagnostic tests, new methods for specimen collection, and new prevalence data in different populations. Public health endorsements for screening are now more specific than ever, yet several important clinical and laboratory questions remain, including questions concerning who to screen and how often to screen. Additional important research is addressing issues related to assay validity, efficacy of screening in different populations, and feasibility of screening on the level of the individual patient and the level of the community and health care system. This article discusses major research findings related to chlamydial screening from the past several years and suggests areas in which additional research is needed. PMID- 12642003 TI - Type-specific Serologic Testing for Herpes Simplex Virus-2. AB - Genital herpes due to infection with herpes simplex virus-2 (HSV-2) affects an estimated 60 million adults in the United States. Over 90% are unaware of their infection but are at risk of transmitting HSV to partners. This ongoing "silent" disease is responsible for the continued increase in HSV prevalence. The recent advent of type-specific serologic tests has allowed accurate differentiation of HSV-1 and HSV-2 infection. Screening of at-risk populations will allow identification of individuals with genital herpes and provide an opportunity for risk reduction counseling and interventions to reduce transmission. PMID- 12642004 TI - Human Herpesvirus 8: Biology and Role in the Pathogenesis of Kaposi's Sarcoma and Other AIDS-related Malignancies. AB - Human herpesvirus type 8, or Kaposi's sarcoma-associated herpesvirus (KSHV), is the only known human g(2) herpesvirus (rhadinovirus) and the most recently discovered tumor virus. KSHV is associated with Kaposi's sarcoma and two other lymphoproliferative disorders in the AIDS setting: primary effusion lymphoma and the plasma cell variant of multicentric Castleman's disease. This review offers an update on the epidemiology and the role of KSHV genes in the development of disease. PMID- 12642006 TI - Vascular abnormalities in salt-sensitive hypertension. PMID- 12642005 TI - Update on the Pathogenesis, Diagnosis, and Therapy of AIDS-related Lymphoma. AB - AIDS-related non-Hodgkin's lymphoma (ARL) is an opportunistic malignancy that foreshortens life more than any other commonly occurring HIV-associated cancer. Treatment strategies include low-dose chemotherapy, chemotherapy given with antiretroviral therapy, and infusional chemotherapy regimens. Rituximab, an anti CD20 monoclonal antibody, has been found to be useful in non-HIV-associated lymphomas and is of interest in ARL as well. Since the advent of highly active antiretroviral therapy, the ARL incidence and survival has changed. This paper reviews the epidemiologic, biologic, and clinical features of ARL with the aim of presenting a cohesive overview of these elements. PMID- 12642007 TI - The association between birth weight and capillary recruitment. PMID- 12642008 TI - The effect of aldosterone on glucose metabolism. AB - There is an association of glucose intolerance and diabetes with primary aldosteronism, but the frequency and mechanisms are not clear. This paper reviews the possible mechanisms of impaired glucose metabolism in primary aldosteronism. Patients with primary aldosteronism can have impaired pancreatic insulin release and reduction in insulin sensitivity. These effects may be due to hypokalemia, but the evidence suggests other factors such as a direct impact of excess aldosterone on insulin action in contributing to the metabolic dysfunction. In general adrenal surgery in cases of aldosterone-producing adenoma will correct the metabolic abnormalities, but it is less sure if treatment with spironolactone in cases of idiopathic hyperplasia will correct impaired glucose tolerance. PMID- 12642009 TI - Angiotensin and insulin resistance: conspiracy theory. AB - Resistance to the metabolic effects of insulin is a contender for the short list of major cardiovascular risk factors. Since the elements of the syndrome of insulin resistance were first articulated together in 1988, numerous epidemiologic investigations and treatment endeavors have established a relationship between the metabolic disarray of impaired insulin action and cardiovascular disease. Angiotensin II, the primary effector of the renin angiotensin system, has also achieved a place in the chronicles of cardiovascular risk factors. Conspiracy mechanisms by which angiotensin II and insulin resistance interact in the pathogenesis of cardiovascular disease are reviewed, with particular attention to recent developments in this engaging area of human research. PMID- 12642010 TI - Insulin resistance in HIV-related lipodystrophy. AB - Lipodystrophy (LD) associated with HIV infection is a syndrome of abnormal fat distribution observed in HIV-infected patients treated with various antiretroviral agents. In addition, insulin resistance and dyslipidemia can occur in HIV-infected patients with or without LD. The demonstration of the latter metabolic disorders in normal subjects using protease inhibitors suggests that these agents could play a causative role in their development independently of HIV status. Possible mechanisms whereby protease inhibitors can hinder insulin actions include inhibition of glucose transporter isoform Glut 4, and altered expression of leptin and tumor necrosis factor-a in adipose tissue. The presence of insulin resistance and dyslipidemia can potentially increase the risk of diabetes, cardiovascular disease, and death. However, short-term data in this regard are inconsistent. Treatment of HIV-related LD with metformin may ameliorate insulin resistance, but its impact on fat redistribution requires additional studies. Temporary cessation of antiretroviral therapy does not appear to reverse body fat changes or insulin resistance, but may partially improve the lipid profile. Further investigations are urgently needed to define the mechanisms and cardiovascular consequences of insulin resistance in HIV-related LD, and to find an effective treatment for this complex syndrome. PMID- 12642011 TI - Aldosterone and specific aldosterone receptor antagonists in hypertension and cardiovascular disease. AB - Recent preclinical and clinical studies indicate that aldosterone, independent of angiotensin II and elevated blood pressure, may play a role in health and disease. In addition to its role in fluid and electrolyte balance and circulatory homeostasis, more recent studies have identified aldosterone as a critical mediator of vascular damage. In animal studies, aldosterone is implicated in cardiac and vascular fibrosis, renal disease, and cerebrovascular damage. These lesions are prevented by specific aldosterone receptor blockade. In clinical studies, aldosterone receptor antagonism is associated with decreased hospitalization, symptomatology, and mortality, and improvement of endothelial dysfunction in patients with chronic heart failure. A better understanding of aldosterone's actions in nonepithelial tissues should pave the way to better protection of organs at risk such as the kidneys, heart, and brain. PMID- 12642012 TI - Contribution of angiotensin-(1-7) to cardiovascular physiology and pathology. AB - Increased understanding of the contribution of angiotensin peptides to the physiologic control of arterial pressure and cardiovascular regulation has been made possible with the introduction of agents that either inhibit the activity of angiotensins forming enzymes or block the action of the peptides at their specific receptor subtypes. This review highlights some of the lessons that have been learned from the study of the actions of angiotensin-(1-7) and its inter relationship with other vasodilator mechanisms that modulate the control systems that determine blood pressure and tissue perfusion. The studies suggest that the renin-angiotensin system acts as a humoral mechanism for blood pressure control through the generation of several distinct forms of angiotensin peptides that may bind to diverse receptor subtypes. PMID- 12642013 TI - Intrarenal angiotensin II and hypertension. AB - Elevations in intrarenal angiotensin II (Ang II) cause reductions in renal function and sodium excretion that contribute to progressive hypertension and lead to renal and vascular injury. Augmentation of intrarenal Ang II occurs by several processes, leading to levels much greater than can be explained from the circulating levels. In Ang II-dependent hypertension, Ang II is internalized via an AT1 receptor mechanism, but there is also sustained intrarenal production of Ang II. Ang II exerts a positive feedback action on intrarenal angiotensinogen (AGT) mRNA and protein. The increased intrarenal AGT production is associated with increased intrarenal and intracellular Ang II contents and urinary AGT excretion rates. The increased urinary AGT indicates spillover of AGT into distal nephron segments supporting enhanced distal Ang II formation and sodium reabsorption. The augmentation of intrarenal Ang II provides the basis for sustained actions on renal function, sodium excretion, and maintenance of hypertension. PMID- 12642014 TI - Role of endothelin-1 in hypertension. AB - Endothelin-1 (ET-1) was first characterized as a potent vasoconstrictor and is overexpressed in the vasculature in different models of hypertension, such as deoxycorticosterone acetate-salt rats, Dahl salt-sensitive rats, and stroke-prone spontaneously hypertensive rats. Moreover, patients with moderate to severe hypertension present increased vascular levels of prepro-ET-1 mRNA. In addition to their blood pressure-lowering effects, ET receptor antagonists are able to reduce vascular growth. Recent data suggest the involvement of an inflammatory response in the effects of ET-1, which contributes to vascular remodeling and endothelial dysfunction. Increasing evidence underscores the potential therapeutic benefit of ET receptor antagonists in different hypertension-related complications, not only in essential hypertension, but also in patients with type 2 diabetes. PMID- 12642015 TI - Inhibition of angiogenesis: a new function for angiotensinogen and des(angiotensin I)angiotensinogen. AB - Angiotensinogen (AGT) can be schematically considered to consist of a combination of an angiotensin I (Ang I) function, located at the N-terminal end, and the presence of a serpin (serine protease inhibitor) structure at the opposite end. des(Ang I)AGT, which accounts for more than 97% of the molecule, apparently has no function. Several serpins (antithrombin, maspin, pigment epithelial-derived factor, and kallistatin) have been recently shown to exert an antiangiogenic activity, suggesting a common mechanism of endothelial cell proliferation and migration. AGT and its renin-cleaved product, des(Ang I)AGT, are also angiogenesis inhibitors, both in vitro and in vivo at concentrations within the range of those observed in plasma. This property most likely results from the structure analogy of AGT with serpins. The pathologic relevance of this new function is still not known, but AGT produced by glial cells may play a role in the stabilization of the blood-brain barrier. These new data must be considered in light of the overall action of the renin-angiotensin system in angiogenesis. PMID- 12642017 TI - The role of angiotensin II-stimulated renal tubular transport in hypertension. AB - The kidney contains a renin-angiotensin system that appears to regulate systemic blood pressure. Angiotensin II (Ang II) has stimulatory effects on sodium transport in multiple nephron segments via binding to plasma membrane AT(1) receptors. In the proximal tubule, Ang II production is substantial. The stimulatory effect of Ang II on proximal sodium transport is enhanced by renal nerves, and is associated with internalization of apical and basolateral receptors. In the cortical collecting duct, AT(1) receptors stimulate transport through apical sodium channels, and in the inner medulla, urea transport is enhanced by Ang II, contributing to increased sodium and water reabsorption. AT(1) receptors may also be linked to increased expression of certain tubular sodium transporters. In contrast to the stimulatory effects of AT(1) receptors on sodium transport, AT(2) receptors expressed in the adult kidney are linked to increased urinary sodium excretion and decreased blood pressure. This suggests that renal tubular AT(1) receptor activation serves as a protective mechanism to increase sodium reabsorption and blood pressure when extracellular fluid volume is threatened, whereas AT(2) receptors dampen this response. The interplay between these two receptor pathways in the kidney could have significant effects on long-term blood pressure control. PMID- 12642016 TI - The role of angiotensin II in regulating vascular structural and functional changes in hypertension. AB - A major hemodynamic abnormality in hypertension is increased peripheral resistance due to changes in vascular structure and function. Structural changes include reduced lumen diameter and arterial wall thickening. Functional changes include increased vasoconstriction and/or decreased vasodilation. These processes are influenced by many humoral factors, of which angiotensin II (Ang II) seems to be critical. At the cellular level, Ang II stimulates vascular smooth muscle cell growth, increases collagen deposition, induces inflammation, increases contractility, and decreases dilation. Molecular mechanisms associated with these changes in hypertension include upregulation of many signaling pathways, including tyrosine kinases, mitogen-activated protein kinases, RhoA/Rho kinase, and increased generation of reactive oxygen species. This review focuses on the role of Ang II in vascular functional and structural changes of small arteries in hypertension. In addition, cellular processes whereby Ang II influences vessels in hypertension are discussed. Finally, novel concepts related to signaling pathways by which Ang II regulates vascular smooth muscle cells in hypertension are introduced. PMID- 12642030 TI - Perspectives in HIV pathogenesis, treatment, and prevention. PMID- 12642018 TI - Angiotensin II and the glomerulus: focus on diabetic kidney disease. AB - The renin-angiotensin system plays a key role in the progression of kidney disease, in addition to its well-described role in the maintenance of extracellular fluid volume and blood pressure. Recent studies have shown that blockade of the renin-angiotensin system at the level of the angiotensin II type 1 receptor can have important effects on proteinuria and the rate of progression of kidney disease in patients with type 2 diabetes mellitus. This review first discusses recent experimental studies relating angiotensin II to kidney function in diabetes mellitus and changes in glomerular permselectivity, and then focuses on recent clinical trials with angiotensin II receptor blockers in patients with type 2 diabetes mellitus. PMID- 12642032 TI - Replication at the level of reverse transcription of HIV-1SF2 strain in Th1 and Th2 cells. AB - It has been reported that Th2 cells better support replication of T-tropic HIV-1 strain than Th1 cells. We investigated at the level of reverse transcription the replication of T-tropic HIV-1SF2 strain in Th1 and Th2 cells. To determine the presence of the early (RU5), intermediate (gag) and full-length (LTR/gag) product of reverse transcription of virus RNA in Th1 and Th2 cells, we employed the PCR technique. Our results revealed that the early (RU5) product of reverse transcription was detectable 30 min after infection in Th2 cells and 1 h after infection in Th1 cells. The product of reverse transcription corresponding to the gag region of HIV-1 was detected in Th2 cells 2 h after infection, whereas the same fragment of viral DNA in Th1 cells was observed 4 h later. The full-length viral DNA was seen in Th2 cells 6 h after infection but the same time course was not detected in the Th1 cells. We also observed that higher efficiency of reverse transcription of viral RNA correlated with better replication of HIV-1SF2 in Th2 than Th1 cells. The higher concentration of p24 HIV-1 antigen in culture medium than cytoplasm of Th1 and Th2 cells confirmed the lack of restriction at the budding step of HIV-1SF2 replication in Th1 and Th2 cells. Our observation may confirm the suggestion that Th0/Th2 cell orientation supports at the reverse transcriptional level the replication of T-tropic HIV-1 strains, which accelerates the onset of AIDS symptoms. PMID- 12642031 TI - Antibodies to Tat and Vpr in the GRIV cohort: differential association with maintenance of long-term non-progression status in HIV-1 infection. AB - The HIV-1 regulatory protein Tat and the accessory protein Vpr are thought to stimulate viral replication and contribute to viral pathogenesis as extracellular proteins. Humoral immune responses to these early viral proteins may therefore be beneficial. We examined serum anti-Tat and anti-Vpr IgG by ELISA in the GRIV cohort of HIV-1 seropositive slow/non-progressors (NP) and fast-progressors (FP), and in seronegative controls. Based on information obtained during a brief follow up period (median = 20 months), NPs were sub-grouped as those maintaining non progression status and therefore stable (NP-S), and those showing signs of disease progression (NP-P). As the primary comparison, initial serum anti-Tat and anti-Vpr IgG (prior to follow-up) were analyzed in the NP sub-groups and in FPs. Anti-Tat IgG was significantly higher in stable NP-S compared to unstable NP-P (P = 0.047) and FPs (P < 0.0005); the predictive value of higher anti-Tat IgG for maintenance of non-progression status was 92% (P = 0.029). In contrast, no difference was observed in anti-Vpr IgG between NP-S and NP-P, although both were significantly higher than FPs (P or =1 in 300) were offered diagnostic amniocentesis or chorionic villus sampling. Of 88 observed Down's syndrome pregnancies, 71 (81%) had a positive screening result (81% detection rate, 95% CI 72-89), and of 46105 unaffected pregnancies, 3200 tested positive (7% false positive rate). These results show that the quadruple test is a better method of screening for Down's syndrome than use of maternal age alone (51% detection rate, 14% false-positive rate) and is more effective than other second trimester screening tests. Therefore, we conclude that the quadruple test should be the test of choice in second trimester screening for Down's syndrome. PMID- 12642054 TI - Tobacco-control treaty language approved despite objections. PMID- 12642053 TI - Alternative lengthening of telomeres and survival in patients with glioblastoma multiforme. AB - Despite advances in the molecular pathogenesis of glioblastoma multiforme, no reliable prognostic markers have been identified. We analysed telomerase activity and telomere lengths in glioblastoma multiformes from 77 patients. 19 patients (25%) had tumours with the alternative-lengthening-of-telomere (ALT) phenotype. Median survival for patients with this phenotype was 542 days (95% CI 114-970) compared with 247 days (224-270) for glioblastoma multiformes with normal telomeres (p=0.0003). Cox's regression analysis showed that this association is independent of age. In patients with non-ALT tumours, telomerase activity did not affect survival (median 287 [199-375] vs 236 [230-242] days, p=0.275). We conclude that ALT is a prognostic indicator for patients with glioblastoma multiforme. PMID- 12642057 TI - EU improves rules for handling human cells and tissue. PMID- 12642058 TI - HIV/AIDS lowers UN's global population estimates. PMID- 12642059 TI - South Africa stalls again on access to HIV drugs. PMID- 12642061 TI - Preventing HIV transmission. PMID- 12642062 TI - New internet brings medicine up to speed. PMID- 12642063 TI - Swiss financial capital attracts suicide tourists. PMID- 12642064 TI - Acute myocardial infarction. AB - Acute myocardial infarction is a common disease with serious consequences in mortality, morbidity, and cost to the society. Coronary atherosclerosis plays a pivotal part as the underlying substrate in many patients. In addition, a new definition of myocardial infarction has recently been introduced that has major implications from the epidemiological, societal, and patient points of view. The advent of coronary-care units and the results of randomised clinical trials on reperfusion therapy, lytic or percutaneous coronary intervention, and chronic medical treatment with various pharmacological agents have substantially changed the therapeutic approach, decreased in-hospital mortality, and improved the long term outlook in survivors of the acute phase. New treatments will continue to emerge, but the greatest challenge will be to effectively implement preventive actions in all high-risk individuals and to expand delivery of acute treatment in a timely fashion for all eligible patients. PMID- 12642065 TI - Prostate cancer epidemiology. AB - Because more and more men are being diagnosed with prostate cancer worldwide, knowledge about and prevention of this disease is important. Epidemiological studies have provided some insight about the cause of prostate cancer in terms of diet and genetic factors. However, compared with other common cancers such as breast and lung cancer, the causes remain poorly understood. Several important issues could help in our understanding of this disease-the variation in incidence of prostate cancer between ethnic populations and the factors leading to familial clustering of the diseases. PMID- 12642067 TI - Children locked away from human rights in the UK. PMID- 12642066 TI - Problems of reporting genetic associations with complex outcomes. AB - Inability to replicate many results has led to increasing scepticism about the value of simple association study designs for detection of genetic variants contributing to common complex traits. Much attention has been drawn to the problems that might, in theory, bedevil this approach, including confounding from population structure, misclassification of outcome, and allelic heterogeneity. Other researchers have argued that absence of replication may indicate true heterogeneity in gene-disease associations. We suggest that the most important factors underlying inability to replicate these associations are publication bias, failure to attribute results to chance, and inadequate sample sizes, problems that are all rectifiable. Without changes to present practice, we risk wastage of scientific effort and rejection of a potentially useful research strategy. PMID- 12642068 TI - The humanitarian cost of a war in Iraq. PMID- 12642069 TI - Homelessness and hardship in Moscow. PMID- 12642070 TI - Worse than a crime; a mistake. PMID- 12642072 TI - Drug advertising in medical journals. PMID- 12642074 TI - Drug advertising in medical journals. PMID- 12642075 TI - Drug advertising in medical journals. PMID- 12642076 TI - Drug advertising in medical journals. PMID- 12642077 TI - Drug advertising in medical journals. PMID- 12642079 TI - Nitroglycerin for septic shock. PMID- 12642080 TI - Philanthropist or opportunist? PMID- 12642081 TI - Medical professionals: a need for introspection. PMID- 12642082 TI - Glucose-induced hypoglycaemia in insulinoma with impaired glucose tolerance. PMID- 12642083 TI - Glucose-induced hypoglycaemia in insulinoma with impaired glucose tolerance. PMID- 12642084 TI - National strategy for suicide prevention in Japan. PMID- 12642085 TI - Tumour marker reference ranges and patients' anxiety. PMID- 12642087 TI - Jonathan Kaplan. PMID- 12642092 TI - Comparison of the basal and glucocorticoid-inducible activities of the upstream regulatory regions of HPV18 and HPV31 in multiple epithelial cell lines. AB - Steroid hormone receptors have been shown to bind to response elements in the upstream regulatory region (URR) of human papillomavirus (HPV) in a ligand dependent manner to affect viral promoter activity. To better understand how the enhancer activity of the URR differs between high risk HPV types, we chose to compare the basal and glucocorticoid-dependent activities of the URRs of HPV18 and HPV31. We found that the URR of HPV18 is a stronger enhancer than the URR of HPV31 in six different cell lines of epithelial origin. Furthermore, the activity of the URR of HPV31 was not inducible by the synthetic glucocorticoid dexamethasone (dex) in any cell line tested, while the URR of HPV18 was dex inducible in the majority of these lines. These studies indicate significant differences between the URRs of high risk HPV types. PMID- 12642093 TI - Suppressor of RNA silencing encoded by Beet yellows virus. AB - Using an Agrobacterium-mediated transient assay, we screened the 15.5-kb genome of the Beet yellows virus for proteins with RNA silencing suppressor activity. Among eight proteins tested, only a 21-kDa protein (p21) was able to suppress double-stranded (ds) RNA-induced silencing of the green fluorescent protein (GFP) mRNA. Restoration of GFP expression by p21 under these conditions had no apparent effect on accumulation of the small interfering RNAs. In addition, p21 elevated the transient expression level of the GFP mRNA in the absence of dsRNA inducer. Similar activities were detected using homologs of p21 encoded by other members of the genus Closterovirus. Computer analysis indicated that p21-like proteins constitute a novel protein family that is unrelated to other recognized suppressors of RNA silencing. Examination of the subcellular distribution in BYV infected plants revealed that p21 is partitioned between soluble cytoplasmic form and proteinaceous inclusion bodies at the cell periphery. PMID- 12642094 TI - Analysis of the role of predicted RNA secondary structures in Ebola virus replication. AB - Thermodynamic modeling of Ebola viral RNA predicts the formation of RNA stem-loop structures at the 3' and 5' termini and panhandle structures between the termini of the genomic (or antigenomic) RNAs. Sequence analysis showed a high degree of identity among Ebola Zaire, Sudan, Reston, and Cote d'Ivoire subtype viruses in their 3' and 5' termini (18 nucleotides in length) and within a second region (internal by approximately 20 nucleotides). While base pairing of the two conserved regions could lead to the formation of the base of the putative stem loop or panhandle structures, the intervening sequence variation altered the predictions for the rest of the structures. Using an in vivo minigenome replication system, we engineered mutations designed to disrupt potential base pairing in the viral RNA termini. Analysis of these variants by screening for enhanced green fluorescent protein reporter expression and by quantitation of minigenomic RNA levels demonstrated that the upper portions of the putative panhandle and 3' genomic structures can be destabilized without affecting virus replication. PMID- 12642095 TI - Rescue of Hantaan virus minigenomes. AB - Hantavirus infections are a major public health concern worldwide. Their widespread geographical distribution and their ability to produce serious, often fatal, human disease underline the need for a system that allows manipulation of these viruses. We describe here the first successful establishment of a reverse genetics technology for Hantaan virus, the prototype of the genus Hantavirus. The system offers a unique opportunity to study the biology of hantaviruses, the pathogenesis of the diseases, and the efficacy of antiviral and prophylactic measures against hantavirus infections. PMID- 12642096 TI - Identification of nucleolus localization signal of betanodavirus GGNNV protein alpha. AB - Betanodavirus greasy grouper (Epinephelus tauvina) nervous necrosis viruses (GGNNV) protein alpha, a virus capsid protein, was detected in both nucleolus and cytoplasm of infected cells of Asian sea bass (SB) and transfected cells of SB and Cos-7 with pcDNA3.1/RNA2. To study its subcellular localization, ORF of protein alpha with 338 aa was fused with enhanced green fluorescent protein (EGFP) gene and was detected in transfected cells in the absence of other viral proteins. In both SB and Cos-7 cells, protein alpha was found to localize EGFP to the nucleolus and cytoplasm. Deletion mutants of protein alpha indicated that N terminal 43 amino acid residues were required to import EGFP-alpha protein into the nucleolus. Further deletions within the 43 amino acid backbone, EGFP/33aa(1 33) and EGFP/30aa(14-43), localized to the nucleolus, suggesting that the 20 amino acids from 14 to 33 of protein alpha were the domain of nucleolus localization. To further determine the nucleolus targeting sequence, deletion mutations within the 20 amino acids of protein alpha were constructed. It was found that the deletion of (23)RRR(25), (29)RRR(31), or (23)RRRANNRRR(31) prevented the accumulation of EGFP fusion proteins into the nucleolus, demonstrating that (23)RRRANNRRR(31) contain the signal required for nucleolar localization. A similar distribution pattern of localization of protein alpha and its deletion mutants in SB and Cos-7 cells suggested that N-terminal residues of protein alpha (23)RRRANNRRR(31) constitute a nucleolus localization signal that functions in both fish and mammalian cells. PMID- 12642097 TI - Absence of Epstein-Barr virus DNA in the tumor cells of European hepatocellular carcinoma. AB - The Epstein-Barr virus (EBV) has recently been associated with hepatocellular carcinoma (HCC) arising in Japanese patients. We analyzed 82 cases of HCC from Germany and the U.K. for the presence of EBV DNA and viral gene products within tumor cells. Initial screening of whole sections using quantitative (Q)-PCR detected EBV DNA in 9/58 U.K. cases and in 9/24 German cases; in positive cases viral load was very low, ranging between 1.4 and 49.1 copies of the EBV genome/1000 cell equivalents, compared to much higher values for EBV-positive Hodgkin's disease and nasopharyngeal carcinoma controls (range, 714-3259/1000 cells). EBV DNA was not detected in the tumor cells of any of the Q-PCR-positive cases either by Q-PCR of pure tumor cell populations isolated by laser capture microdissection or by isotopic in situ hybridization. Furthermore, none of the German or U.K. HCC tumors tested positive for EBER or EBNAI expression in tumor cells. Our results provide strong evidence that HCCs from the U.K. or Germany are not associated with EBV. PMID- 12642098 TI - Nuclear export of influenza viral ribonucleoprotein is temperature-dependently inhibited by dissociation of viral matrix protein. AB - The influenza virus copies its genomic RNA in the nuclei of host cells, but the viral particles are formed at the plasma membrane. Thus, the export of new genome from the nucleus into the cytoplasm is essential for viral production. Several viral proteins, such as nucleoprotein (NP) and RNA polymerases, synthesized in the cytoplasm, are imported into the nucleus, and form viral ribonucleoprotein (vRNP) with new genomic RNA. vRNP is then exported into the cytoplasm from the nucleus to produce new viral particles. M1, a viral matrix protein, is suggested to participate in the nuclear export of vRNP. It was found unexpectedly that the production of influenza virus was suppressed in MDCK cells at 41 degrees C, although viral proteins were synthesized and the cytopathic effect was observed in host cells. Indirect immunofluorescent staining with anti-NP or M1 monoclonal antibody showed that NP and M1 remained in the nuclei of infected cells at 41 degrees C, suggesting that a suppression of viral production was caused by inhibition of the nuclear export of these proteins. The cellular machinery for nuclear export depending on CRM1, which mediates the nuclear export of influenza viral RNP, functioned normally at 41 degrees C. Glycerol-density gradient centrifugation demonstrated that vRNP also formed normally at 41 degrees C. However, an examination of the interaction between vRNP and M1 by immunoprecipitation indicated that M1 did not associate with vRNP at 41 degrees C, suggesting that the association is essential for the nuclear export of vRNP. Furthermore, when infected cells incubated at 41 degrees C were cultured at 37 degrees C, the interaction between vRNP and M1 was no longer detected even at 37 degrees C. The results suggest that M1 synthesized at 41 degrees C is unable to interact with vRNP and the dissociation of M1 from vRNP is one of the reasons that the transfer of vRNP into the cytoplasm from the nucleus is prevented at 41 degrees C. PMID- 12642099 TI - Distribution of the attachment (G) glycoprotein and GM1 within the envelope of mature respiratory syncytial virus filaments revealed using field emission scanning electron microscopy. AB - Field emission scanning electron microscopy (FE SEM) was used to visualize the distribution of virus-associated components, the virus-attachment (G) protein, and the host-cell-derived lipid, GM1, in respiratory syncytial virus (RSV) filaments. RSV-infected cells were labeled in situ with a G protein antibody (MAb30) whose presence was detected using a second antibody conjugated to colloidal gold. No bound MAb30 was detected in mock-infected cells, whereas significant quantities bound to viral filaments revealing G protein clusters throughout the filaments. GM1 was detected using cholera toxin B subunit conjugated to colloidal gold. Mock-infected cells revealed numerous GM1 clusters on the cell surface. In RSV-infected cells, these gold clusters were detected on the filaments in low, but significant, amounts, indicating the incorporation of GM1 within the viral envelope. This report describes the first use of FE SEM to map the distribution of specific structural components within the envelope of a Paramyxovirus. PMID- 12642100 TI - The P4 promoter of the parvovirus minute virus of mice is developmentally regulated in transgenic P4-LacZ mice. AB - Activation of the minute virus of mice (MVM) P4 promoter is a key step in the life cycle of the virus and is completely dependent on host transcription factors. Since transcription-factor composition varies widely in different cell types, there is the possibility that only some cell types in the host organism have the capacity to initiate expression from the P4 promoter and therefore that the promoter may be a factor in determining the tropism of MVM. In this study, the ability of various cell types to activate P4, independent of the other virus host interactions, was examined in transgenic mouse lines bearing a beta galactosidase reporter sequence driven by the P4 promoter. It was found that lacZ was expressed during embryogenesis and in the adult in a cell-type-specific and differentiation-dependent pattern. The data are consistent with cell-type and stage-specific activation of the P4 promoter having a role in determining the host cell-type range of MVM. The ability of some parvoviruses to replicate in, and kill oncogenically transformed cells, and to destroy induced tumors in laboratory animals is the basis of recent approaches to use MVM-based vectors in cancer gene therapy. Since these vectors rely on the activation of the P4 promoter by the target tissues, understanding the promoter dependence on cell type and differentiation status is important for their design and potential use. PMID- 12642102 TI - Plasma membrane localization and fusion inhibitory activity of the cowpox virus serpin SPI-3 require a functional signal sequence and the virus encoded hemagglutinin. AB - The cowpox virus (CPV) glycoprotein serpin SPI-3, a functional protease inhibitor, and the viral hemagglutinin (HA) are required to prevent fusion of wt CPV infected cells. SPI-3 and HA from CPV infected cells co-localize to the plasma membrane and are found in extracellular enveloped virus (EEV). We also show that an N-terminal SPI-3 signal sequence, but not glycosylation, is required for membrane localization and fusion inhibition. In the absence of HA (CPVDeltaHA), no SPI-3 is found on the membrane and infected cells fuse. Conversely, HA from both wt CPV and CPVDeltaSPI-3 infections is on the membrane, indicating a requirement of HA for SPI-3 plasma membrane localization. In the absence of HA, secretion of SPI-3 or SPI-3 N-glyc(-) was markedly enhanced, suggesting HA serves to retain SPI-3 on the plasma membrane,thereby preventing cell fusion. PMID- 12642101 TI - Effects of the cowpea chlorotic mottle bromovirus beta-hexamer structure on virion assembly. AB - The X-ray crystal structure of Cowpea chlorotic mottle bromovirus (CCMV) revealed a unique tubular structure formed by the interaction of the N-termini from six coat protein subunits at each three-fold axis of the assembled virion. This structure, termed the beta-hexamer, consists of six short beta-strands. The beta hexamer was postulated to play a critical role in the assembly and stability of the virion by stabilizing hexameric capsomers. Mutational analyses of the beta hexamer structure, utilizing both in vitro and in vivo assembly assays, demonstrate that this structure is not required for virion formation devoid of nucleic acids in vitro or for RNA-containing virions in vivo. However, the beta hexamer structure does contribute to virion stability in vitro and modulates disease expression in vivo. These results support a model for CCMV assembly through pentamer intermediates. PMID- 12642103 TI - Induction of apoptosis in frog virus 3-infected cells. AB - The ability of frog virus 3 (FV3), the type species of the family Iridoviridae, to induce apoptosis was examined by monitoring DNA cleavage, chromatin condensation, and cell-surface expression of phosphotidylserine (PS) in fathead minnow (FHM) and baby hamster kidney (BHK) cells. In productively infected FHM cells, DNA fragmentation was first noted at 6-7 h postinfection and was clearly seen by 17 h postinfection, while chromatin condensation was detected at 8.5 h postinfection. As with some other viruses, FV3-induced apoptosis did not require de novo viral gene expression as both heat-inactivated and UV-inactivated virus readily triggered DNA fragmentation in FHM cells. Moreover, FV3-induced apoptosis was blocked in FHM cells by the pan-caspase inhibitor Z-VAD-FMK, suggesting that virus infection triggers programmed cell death through activation of the caspase cascade. FV3 infection also triggered apoptosis in BHK cells as monitored by TUNEL and annexin V binding assays. To determine whether FV3, similar to other large DNA viruses, encoded proteins that block or delay apoptosis, mock- and FV3 infected FHM cells were osmotically shocked and assayed for DNA fragmentation 3 hours later. DNA fragmentation was clearly seen whether or not shocked cells were previously infected with FV3, indicating that infection with FV3 did not block apoptosis induced by osmotic shock in FHM cells. The above results demonstrate that iridoviruses triggered apoptosis and that the induction of programmed cell death did not require viral gene expression. However, it remains to be determined if virion attachment to target cells is sufficient to induce cell death, or if apoptosis is triggered directly or indirectly by one or more virion-associated proteins. PMID- 12642104 TI - Regulation of MSV and WDV virion-sense promoters by WDV nonstructural proteins: a role for their retinoblastoma protein-binding motifs. AB - In this work we demonstrate that wheat dwarf virus (WDV) RepA can activate WDV and maize streak virus (MSV) virion (V)-sense expression in plant tissues. Rep alone does not have any effect on the silent WDV promoter and it represses the basal MSV promoter activity. MSV promoter activation by RepA depends on an intact RepA retinoblastoma protein (RB)-binding domain. Promoter repression by Rep also depends on this domain to some extent. Mutation of the RepA RB-binding domain has no effect on WDV promoter activation. The WDV promoter contains two sites that fit the consensus E2F-binding site. One, WDV1, binds human E2F-1 in one-hybrid assays in yeast. It also binds specifically to maize and wheat proteins in vitro and, when fused to a minimal 35S promoter, it confers responsiveness to RepA only when the RepA RB-binding domain and the WDV1 site are intact. In the whole WDV V sense promoter context, mutations of this sequence have no effect, suggesting that additional sequences are important for RepA-mediated promoter activation. PMID- 12642105 TI - Distribution of TT virus (TTV), TTV-like minivirus, and related viruses in humans and nonhuman primates. AB - TT virus (TTV) and TTV-like minivirus (TLMV) are small DNA viruses with single stranded, closed circular, antisense genomes infecting man. Despite their extreme sequence heterogeneity (>50%), a highly conserved region in the untranslated region (UTR) allows both viruses to be amplified by polymerase chain reaction (PCR). TTV/TLMV infection was detected in 88 of 100 human plasma samples; amplified sequences were differentiated into TTV and TLMV by analysis of melting profiles, showing that both viruses were similarly prevalent. PCR with UTR primers also detected frequent infection with TTV/TLMV-related viruses in a wide range of apes (chimpanzees, gorillas, orangutans, gibbons) and African monkey species (mangabeys, drills, mandrills). These findings support the hypothesis for the co-evolution of TTV-like viruses with their hosts over the period of primate speciation, potentially analogous to the evolution of primate herpesviruses. PMID- 12642106 TI - Comparative histopathological studies in the early stages of acute pathogenic and nonpathogenic SHIV-infected lymphoid organs. AB - To clarify the early pathological events in simian and human immunodeficiency chimeric virus (SHIV)-infected lymphoid organs, we examined rhesus macaques infected with an acute pathogenic SHIV (SHIV89.6P) or a nonpathogenic SHIV (NM 3rN) by sequential biopsies and serial necropsies. In the SHIV89.6P-infected monkeys, acute thymic involution as shown by increased cortical tingible-body macrophages and by neutrophilic infiltrates without follicular aggregation in the medulla began within 14 days postinoculation (dpi). Cells that were strongly positive for the virus were identified in the thymic medulla. SHIV89.6P-infected lymph nodes showed severe paracortical lymphadenitis with scattered virus positive cells at 14 dpi and they developed paracortical depletion without the obvious follicular involution. In contrast, NM-3rN-infected monkeys showed no signs of thymic dysinvolution and the lymph nodes exhibited only follicular hyperplasia. NM-3rN-infected monkeys showed much fewer virus-positive cells in these lymphoid tissues than did SHIV89.6P-infected monkeys during the same period. These differences clearly reflect the difference in the virulence of these SHIVs. PMID- 12642107 TI - Positive- and negative-acting signals combine to determine differential RNA replication from the paramyxovirus simian virus 5 genomic and antigenomic promoters. AB - The cis-acting signals found at the 3' ends of the genomic and antigenomic RNAs are a major factor determining the level of paramyxovirus RNA replication from each promoter. Using a minigenome system that reconstitutes SV5 RNA synthesis from cDNA-derived components, we show here that the genomic promoter (GP) for the paramyxovirus SV5 directs RNA replication approximately 14-fold lower than that seen from the antigenomic promoter (AGP). The goal of this study was to identify cis-acting signals responsible for differential levels of RNA replication from the SV5 GP and AGP. We have previously shown that the SV5 AGP contains three sequence-dependent elements (CRI, CRII, and Region III) that are separated by sequence-independent spacer regions. Minigenomes containing chimeric promoters were constructed to test the hypothesis that transfer of discrete cis-acting AGP elements to the GP could confer higher replication properties to the GP. Minigenomes containing a substitution of the AGP CRI, CRII, or Region III elements alone in place of the corresponding GP sequences did not show enhanced levels of RNA replication. However, transfer of both the AGP 3' terminal CRI and Region III elements into the corresponding sites of the GP led to a minigenome which replicated to approximately 40% of the levels seen with the AGP. This enhanced RNA replication from the GP was further increased up to AGP levels by also including the intervening AGP segment (bases 20-50) located between CRI and Region III. Importantly, transfer of nonviral sequences in place of GP bases 20 50 also increased RNA replication to levels approaching that of the AGP, but only in the context of the AGP CRI and Region III substitutions. These data indicate that differential levels of RNA replication from the SV5 GP and AGP are due to a combination of positive-acting signals in the AGP (CRI and Region III) and a negative-acting signal in the GP (bases 20-50). Possible functions for the SV5 promoter elements in determining RNA replication levels are proposed. PMID- 12642108 TI - Providence virus: a new member of the Tetraviridae that infects cultured insect cells. AB - We identified a new member of the Tetraviridae, Providence virus (PrV), persistently infecting a midgut cell line derived from the corn earworm (Helicoverpa zea). Virus purified from these cells also productively infected a H. zea fat body cell line, and a cell line from whole embryos of the beet armyworm, Spodoptera exigua. PrV is thus the first tetravirus shown to replicate in cell culture. PrV virions are isometric particles composed of two structural proteins (60 and 7.4 kDa) that encapsidate both the genomic (6.4 kb) and the subgenomic (2.5 kb) RNAs. The monopartite organization of the PrV genome resembles that of Nudaurelia beta virus and Thosea asigna virus, members of the genus Betatetravirus. The predicted sequence of the PrV structural proteins demonstrates homology to tetraviruses in both genera. The infectivity of PrV for cultured cells uniquely permitted examination of tetravirus RNA and protein synthesis during synchronous infection. The discovery of PrV greatly facilitates studies of tetravirus molecular biology. PMID- 12642109 TI - Chimeric immune receptor T cells bypass class I requirements and recognize multiple cell types relevant in HIV-1 infection. AB - Transduction of T cells with a chimeric immune T cell receptor (CIR) has been proposed as a strategy to generate cellular immunity against viral pathogens such as HIV-1. In the case of the CD4-CD3-zeta chain (CD4-zeta) CIR, specificity for HIV-1 is conferred by binding of the CD4 moiety to gp120 on the surface of infected cells. However, it is unclear whether CD4-zeta-T cells may differ from naturally derived CD8(+) cytotoxic T cells (CTL) in their susceptibility to viral escape mechanisms or ability to recognize different cell types that support viral replication. We demonstrate that CIR-T cells can mediate antiviral activity against HIV-1 in cells that are resistant to class I-restricted CTL-mediated activity. Furthermore, CIR-T cells can suppress virus in multiple cell types, including monocytes, dendritic cells, and lymphocyte-dendritic cell clusters. These results provide evidence that T cells can be redirected against novel targets, and that independence from the class I pathway may have distinct advantages. PMID- 12642110 TI - Memory T cells protect chicks from acute infectious bronchitis virus infection. AB - Infectious bronchitis has remained one of the most difficult to control diseases in poultry since it was first described in 1931. Previous studies demonstrated that primary CD8(+) T lymphocytes collected at 10 days post-infection (p.i.) are important in controlling acute infection. To further investigate the role of memory T cells in protection, T lymphocytes collected from B19/B19 chicken spleens at 2, 3, 4, and 6 weeks p.i. were transferred to six-day-old syngeneic chicks one day prior to challenging with 10(6) EID(50) of the IBV Gray strain. Memory immune T cells collected at 3 to 6 weeks p.i. provided dose responsive protection from clinical illness. The greatest protection was observed after the transfer of 10(7) T cells collected at 6 weeks p.i., whereas T cells collected at 2 weeks p.i. did not protect. Annexin-V staining of the spleen cells demonstrated that the cells collected at 2 weeks p.i. were undergoing significantly more apoptosis than cells collected at 10 days p.i. Specific antibody production in sera collected at 7 days p.i. did not correlate with protection. T cell subtype depletion demonstrated that CD8(+), not CD4(+), T cells were critical. Memory T cells can be detected in peripheral blood mononuclear cells up to at least 10 weeks p.i. These results demonstrated that IBV specific CD8(+) memory T cells generated at 3 to 6 weeks p.i. can protect syngeneic chicks from acute IBV infection. PMID- 12642111 TI - Constraints-based models: regulation of gene expression reduces the steady-state solution space. AB - Constraints-based models have been effectively used to analyse, interpret, and predict the function of reconstructed genome-scale metabolic models. The first generation of these models used "hard" non-adjustable constraints associated with network connectivity, irreversibility of metabolic reactions, and maximal flux capacities. These constraints restrict the allowable behaviors of a network to a convex mathematical solution space whose edges are extreme pathways that can be used to characterize the optimal performance of a network under a stated performance criterion. The development of a second generation of constraints based models by incorporating constraints associated with regulation of gene expression was described in a companion paper published in this journal, using flux-balance analysis to generate time courses of growth and by-product secretion using a skeleton representation of core metabolism. The imposition of these additional restrictions prevents the use of a subset of the extreme pathways that are derived from the "hard" constraints, thus reducing the solution space and restricting allowable network functions. Here, we examine the reduction of the solution space due to regulatory constraints using extreme pathway analysis. The imposition of environmental conditions and regulatory mechanisms sharply reduces the number of active extreme pathways. This approach is demonstrated for the skeleton system mentioned above, which has 80 extreme pathways. As regulatory constraints are applied to the system, the number of feasible extreme pathways is reduced to between 26 and 2 extreme pathways, a reduction of between 67.5 and 97.5%. The method developed here provides a way to interpret how regulatory mechanisms are used to constrain network functions and produce a small range of physiologically meaningful behaviors from all allowable network functions. PMID- 12642112 TI - Threat displays are not handicaps. AB - Within a general framework of handicap signalling it was proposed that threat displays are handicaps, they can work only if they put the signaller at a disadvantage, which is only acceptable to honest signallers. The aim of the present article is to investigate this proposal with the help of a simple game theoretical model. It was found: (1) that the use of cost-free signals is an ESS against the invasion of handicapped signals even if cheating is played as part of a mixed strategy in the population; (2) that the use of handicaps may be an ESS against cost-free signals but only if we assume that the invading cost-free signal is not accepted by weak individuals as a signal of strength; (3) that the establishment of a handicapped signal in the first place is an unresolved problem, because both cost free signals and negative-handicaps are evolutionarily stable against the invasion of handicaps; (4) that in contrast to handicaps the use of negative-handicaps can invade a population using cost-free signals (a negative-handicap is a signal which may serve other functions as well); (5) that negative-handicaps are ESS against cost-free signals as well as against handicaps; and (6) thus, the most likely evolutionary end point is that the biggest negative-handicap would be used as a threat display. This is a posture, which prepares the animal most efficiently to fight; hence, most probably it is the initial position of the fighting technique of the given species. (7) Finally, the investigation of the threat displays of well-studied taxa (great tit, cats, dogs, and hoofed mammals) confirms that threat displays are indeed negative handicaps. They do not put the user into a disadvantaged position, instead the initial position of the species specific fighting technique is used as a threat display as predicted by the present model. PMID- 12642113 TI - Modelling the dynamics of LCMV infection in mice: II. Compartmental structure and immunopathology. AB - In this study, we develop a mathematical model for analysis of the compartmental aspects and immunopathology of lymphocytic choriomeningitis virus (LCMV) infection in mice. We used sets of original and published data on systemic (extrasplenic) virus distribution to estimate the parameters of virus growth and elimination for spleen and other anatomical compartments, such as the liver, kidney, thymus and lung as well as transfer rates between blood and the above organs. A mathematical model quantitatively integrating the virus distribution kinetics in the host, the specific cytotoxic T lymphocyte (CTL) response in spleen and the re-circulation of effector CTL between spleen, blood and liver is advanced to describe the CTL-mediated immunopathology (hepatitis) in mice infected with LCMV. For intravenous and "peripheral" routes of infection we examine the severity of the liver disease, as a function of the virus dose and the host's immune status characterized by the numbers of precursor and/or cytolytic effector CTL. The model is used to predict the efficacy of protection against virus persistence and disease in a localized viral infection as a function of the composition of CTL population. The modelling analysis suggests quantitative demands to CTL memory for maximal protection against a wide range of doses of infection with a primarily peripheral site of virus replication without the risk of favoring immunopathology. It specifies objectives for CTL vaccination to ensure virus elimination with minimal immunopathology vs. vaccination for disease. PMID- 12642114 TI - Modeling genetic switches with positive feedback loops. AB - In this paper, we develop a new methodology to design synthetic genetic switch networks with multiple genes and time delays, by using monotone dynamical systems. We show that the networks with only positive feedback loops have no stable oscillation but stable equilibria whose stability is independent of the time delays. In other words, such systems have ideal properties for switch networks and can be designed without consideration of time delays, because the systems can be reduced from functional spaces to Euclidian spaces. Therefore, we can ensure that the designed switches function correctly even with uncertain delays. We first prove the basic properties of the genetic networks composed of only positive feedback loops, and then propose a procedure to design the switches, which drastically simplifies analysis of the switches and makes theoretical analysis and design tractable even for large-scaled systems. Finally, to demonstrate our theoretical results, we show biologically plausible examples by designing a synthetic genetic switch with experimentally well investigated lacI, tetR, and cI genes for numerical simulation. PMID- 12642115 TI - Genome size and operon content. AB - Prokaryotic genes are often organized into operons, clusters of genes that are transcribed together. Because all genes in an operon must be transcribed in the same direction, this organization will be reflected in a tendency for nearby genes to have the same orientation. This tendency can be used to estimate the degree to which the genes in a genome are clustered into operons. Application of the technique to Escherichia coli yields results that are similar to estimates based on detailed examination of the genome and empirical knowledge about particular operons. Results for Saccharomyces cerevisiae are consistent with the near absence of polycistronic transcripts in eukaryotes. The method is easily applied to other genomes that have been sequenced and annotated. Analysis of 26 bacterial and archaeal genomes indicates that the degree of clustering varies widely among prokaryotes. Comparison of these genomes shows that those containing more genes tend to have less clustering of genes into operons. This observation may have implications concerning the evolution of operons. PMID- 12642116 TI - Facultative sexual reproduction under frequency-dependent selection on a single locus. AB - The evolution of a facultative sexual strategy that simultaneously produced sexual and asexual individuals was studied theoretically, under negative frequency-dependence of fitness. The organism was considered to be diploid, characterized by two loci concerning fitness and determining sexual strategy, between which a certain degree of linkage existed. The locus concerning fitness was assumed to involve two alleles, resulting in three genotypes, the relative fitness of an individual being defined by a decreasing function of frequency of its own genotype on this locus in the population. The sexual reproductive strategy was considered to be determined by three alleles; asexual, obligate sexual and facultative sexual. Simulations under various linkages between loci and level of frequency dependence of fitness showed that a facultative sexual strategy was generally able to invade and increase in the population. In particular, when the level of frequency dependence was high to some degree, the facultative strain producing many sexual individuals tended to exclusively occupy the population. Namely, the frequency-dependent selection resulted in a predominance of obligate sexual strategy over asexual strategy, simultaneously causing a subordination of the former to the facultative sexual strategy. This indicated that the evolution of sex should be considered carefully with respect to the possibility of invasion of facultative sex. PMID- 12642117 TI - The universal ancestor was a thermophile or a hyperthermophile: tests and further evidence. AB - The existence of a correlation between the optimal growth temperature of various organisms and a thermophily index (based on the propensity of amino acids to enter more frequently into the proteins of thermophiles/hyperthermophiles) allows inferences to be made on the mesophilic or thermophilic nature of the last universal common ancestor (LUCA). By reconstructing the ancestral sequences of the various ancestors using methods based on maximum likelihood and maximum parsimony, these sequences can be attributed to the mesophiles or (hyper)thermophiles and the following conclusions can be drawn. (1) There is no evidence that the LUCA might have been a mesophile and observations seem to imply that the LUCA was a thermophile or a hyperthermophile; (2) The ancestors of the Archaea and Bacteria domains seem to be (hyper)thermophiles while that of the Eukarya domain turns out to be a mesophile. These conclusions are independent of both (i) where the root is located on the topology of the universal tree (based on that of the small subunit ribosomal RNA) and (ii) the presence of hyperthermophile bacteria near the node of the Bacteria domain ancestor. These conclusions are easier to interpret in the light of the hypotheses that see the origin of life taking place at a high temperature. PMID- 12642118 TI - A simple model for the arterial system. AB - We present a simple model for the arterial part of the cardiovascular system, based on Poiseuille flow constrained by the power dissipated into the cells lining the vessels. This, together with the assumption of a volume-filling network, leads to correct predictions for the evolution of vessel radii, vessel lengths and blood pressure in the human arterial system. The model can also be used to find exponents for allometric scaling, and gives good agreement with data on mammals. PMID- 12642119 TI - Language dynamics in finite populations. AB - Any mechanism of language acquisition can only learn a restricted set of grammars. The human brain contains a mechanism for language acquisition which can learn a restricted set of grammars. The theory of this restricted set is universal grammar (UG). UG has to be sufficiently specific to induce linguistic coherence in a population. This phenomenon is known as "coherence threshold". Previously, we have calculated the coherence threshold for deterministic dynamics and infinitely large populations. Here, we extend the framework to stochastic processes and finite populations. If there is selection for communicative function (selective language dynamics), then the analytic results for infinite populations are excellent approximations for finite populations; as expected, finite populations need a slightly higher accuracy of language acquisition to maintain coherence. If there is no selection for communicative function (neutral language dynamics), then linguistic coherence is only possible for finite populations. PMID- 12642120 TI - Veno-occlusive disease: cytokines, genetics, and haemostasis. AB - Hepatic veno-occlusive disease (VOD) is a major cause of morbidity and mortality following high dose cytotoxic therapy for stem cell transplantation (SCT). Pre existing liver damage, SCT-related therapy, and genetic polymorphisms all appear to increase the risk of developing VOD. Studies of biological markers during SCT suggest that cytokines, haemostasis, and hepatic drug metabolism via the glutathione pathway are all involved in the pathogenesis of VOD. Until recently, treatment options were limited and experimental therapies directed at the pathogenesis of the disease were mostly unsuccessful. However, Defibrotide, a relatively new agent that has modulatory effects on vascular endothelium, cytokine release, and haemostasis, has been used with some success in the management and prophylaxis of VOD. In the future, a better understanding of genetic polymorphisms and biological markers which may be important in the pathogenesis of VOD, may enable us to predict which patients are most likely to be affected. PMID- 12642121 TI - The molecular pathogenesis of acute promyelocytic leukaemia: implications for the clinical management of the disease. AB - Acute promyelocytic leukaemia (APL) is characterised by chromosomal rearrangements of 17q21, leading to fusion of the gene encoding retinoic acid receptor alpha (RARalpha) to a number of alternative partner genes (X), the most frequent of which are PML (>95%), PLZF (0.8%) and NPM (0.5%). Over the last few years, it has been established that the X-RARalpha fusion proteins play a key role in the pathogenesis of APL through recruitment of co-repressors and the histone deacetylase (HDAC)-complex to repress genes implicated in myeloid differentiation. Paradoxically, the X-RARalpha fusion protein has the potential to mediate myeloid differentiation at pharmacological doses of its ligand (all trans-retinoic acid (ATRA)), which is dependent on the dissociation of the HDAC/co-repressor complex. Arsenic compounds have also been shown to be promising therapeutic agents, leading to differentiation and apoptosis of APL blasts. It is now apparent that the nature of the RARalpha-fusion partner is a critical determinant of response to ATRA and arsenic, underlining the importance of cytogenetic and molecular characterisation of patients with suspected APL to determine the most appropriate treatment approach. Standard protocols involving ATRA combined with anthracycline-based chemotherapy, lead to cure of approximately 70% patients with PML-RARalpha-associated APL. Patients at high risk of relapse can be identified by minimal residual disease monitoring. The challenge for future studies is to improve complete remission rates through reduction of induction deaths, particularly due to haemorrhage, identification of patients at high risk of relapse who would benefit from additional therapy, and identification of a favourable-risk group, for which treatment intensity could be reduced, thereby reducing risks of treatment toxicity and development of secondary leukaemia/myelodysplasia. With the advent of ATRA and arsenic, APL has already provided the first example of successful molecularly targeted therapy; it is hoped that with further understanding of the pathogenesis of the disease, the next decade will yield further improvements in the outlook for these patients. PMID- 12642122 TI - The role of hydroxyurea in the management of sickle cell disease. AB - Sickle cell disease (SCD) is one of the most common genetic diseases with some 250,000 new births each year. Most patients suffer intermittent pain crises and life-threatening events while life expectancy is considerably reduced. Until the last decade management was purely preventative or supportive aimed at symptom control. Apart from stem cell transplant, there is no cure but the oral chemotherapeutic drug hydroxyurea (HU) has now established a role in ameliorating the disease and improving life expectancy for most patients. There are side effects and risks of HU treatment in SCD but for moderate and severely affected patients, the benefits can be significant. PMID- 12642123 TI - Safety of the blood supply: role of pathogen reduction. AB - Even though the blood supply is very safe, the risk of transfusion transmitted disease is not zero. To improve the safety of the blood supply, pathogen reduction (PR) technology has been developed. The principle of most current PR strategies involves modifying DNA or RNA templates and making them inaccessible to DNA or RNA polymerase. Several platforms of pathogen reduction are available including psoralens, alkylating compounds, binary ethyleneimine-like compounds, riboflavin, methylene blue, and solvent-detergent treatment. PR systems have been designed for RBC, plasma, and platelets. PR technology has been found to be effective for a variety of pathogens including lipid-enveloped and non-enveloped viruses, bacteria and parasites. Pre-clinical studies and Phase III clinical trials to evaluate the efficacy and safety of these PR technologies are currently ongoing. PMID- 12642124 TI - Carbonylbisphosphonate and (diazomethylene)bisphosphonate analogues of AZT 5(') diphosphate. AB - A novel nucleotide analogue is described, in which the alpha,beta-phosphoric anhydride oxygen of a nucleoside 5(')-diphosphate is replaced by a carbonyl group: the carbonylbisphosphonate analogue 5 of 2('),3(')-dideoxy-3(') azidothymidine 5(')-diphosphate (AZT 5(')-diphosphate). 5 was synthesized from tetramethyl (diazomethylene)bisphosphonate 1 via the trimethyl ester 4 of the corresponding AZT 5(')-(diazomethylene)bisphosphonate 6, which is also a new type of nucleotide analogue. The ultimate product 5 was isolated by reverse-phase HPLC, and characterized by 31P, 13C, and 1H NMR; and by high-resolution mass spectrometry. The ketone group of 5 is a visible chromophore (yellow) and reversibly forms a colorless hydrate. The ketone hydrate 'pK' is about 4.2 when excess of magnesium ion is present. The potential of such analogues as novel inhibitors of enzymes mediating nucleotide-dependent biochemical processes is discussed. PMID- 12642126 TI - Metal ions and the stereochemistry of ribozyme reactions. AB - Inversion of configuration at phosphorus during ribozyme-catalyzed cleavage of RNA is usually considered unequivocal proof of in-line attack, but the relevant pseudorotation diagram for formation of the 2('),3(')-cyclic phosphate shows that inversion is not inconsistent with adjacent attack as long as breakdown of the trigonal bipyramid is in-line. For the reaction to occur by adjacent attack, a normally unstable apical oxyanion in the trigonal bipyramidal intermediate would have to be stabilized. Density-functional calculations show that a metal ion such as magnesium could perform this stabilization. We conclude that the possibility of adjacent attack should not be too hastily dismissed in cases where the setup is closer to adjacent than to in-line geometry. PMID- 12642125 TI - Solution structure and design of dithiophosphate backbone aptamers targeting transcription factor NF-kappaB. AB - A variety of monothio- and dithiosubstituted duplex aptamers targeting NF-kappaB have been synthesized and designed. The specificity and affinity of the dithioate aptamers of p50 and RelA(p65) NF-kappaB homodimers was determined by gel shift experiments. The NMR solution structures for several unmodified and dithioate backbone modified 14-base paired duplex aptamers have been determined by a hybrid, complete matrix (MORASS)/restrained molecular dynamics method. Structural perturbations of the dithioate substitutions support our hypothesis that the dithioate binds cations less tightly than phosphoryl groups. This increases the electrostatic repulsion across the B-form narrow minor groove and enlarges the minor groove, similar to that found in A-form duplexes. Structural analysis of modeled aptamer complexes with NF-kappaB homo- and heterodimers suggests that the dithioate backbone substitution can increase the aptamer's relative affinity to basic groups in proteins such as NF-kappaB by helping to "strip" the cations from the aptamer backbone. PMID- 12642127 TI - Bicelles in structure-function studies of membrane-associated proteins. AB - Bicelles are a novel form of long-chain/short-chain phospholipid aggregates, which are useful for biophysical and biochemical studies of membrane-associated biomolecules. In this work, we review the development of bicelles and their uses in structural characterization (primarily via NMR, circular dichroism, and fluorescence) of membrane-associated peptides. We also show that bicellar phospholipids are substrates for lipolytic enzymes. For this latter work, we employed a 31P NMR enzymatic assay system to examine the kinetic behavior of cobra venom phospholipase A(2) toward a variety of bicellar substrates. This enzyme hydrolyzed all bicelle lipids at rates comparable to those found for the enzyme action on traditional micellar substrates, which are the best substrates for this enzyme. In addition, we found that this PLA(2) showed no significant preference for long-chain or short-chain phospholipids when they were presented as mixtures in bicelles. PMID- 12642128 TI - A novel mechanism for inhibition of HIV-1 reverse transcriptase. AB - The human immunodeficiency virus (HIV) epidemic is an important medical problem. Although combination drug regimens have produced dramatic decreases in viral load, current therapies do not provide a cure for HIV infection. We have used structure-based design and combinatorial medicinal chemistry to identify potent and selective HIV-1 reverse transcriptase (RT) inhibitors that may work by a mechanism distinct from that of current HIV drugs. The most potent of these compounds (compound 4, 2-naphthalenesulfonic acid, 4-hydroxy-7-[[[[5-hydroxy-6 [(4-cinnamylphenyl)azo]-7-sulfo-2-naphthalenyl]amino]carbonyl]amino]-3-[(4 cinnamylphenyl)azo], disodium salt) has an IC(50) of 90 nM for inhibition of polymerase chain extension, a K(d) of 40 nM for inhibition of DNA-RT binding, and an IC(50) of 25-100 nM for inhibition of RNaseH cleavage. The parent compound (1) was as effective against 10 nucleoside and non-nucleoside resistant HIV-1 RT mutants as it was against the wild-type enzyme. Compound 4 inhibited HIV-1 RT and murine leukemia virus (MLV) RT, but it did not inhibit T(4) DNA polymerase, T(7) DNA polymerase, or the Klenow fragment at concentrations up to 200 nM. Finally, compound 4 protected cells from HIV-1 infection at a concentration more than 40 times lower than the concentration at which it caused cellular toxicity. PMID- 12642129 TI - Managing the mutations of an old threat. PMID- 12642140 TI - Carl Linnaeus: pictures and propaganda. AB - How do scientists become famous? Carl Linnaeus was an expert at self-promotion who used portraits to gain patronage and consolidate his reputation. His sexualized classification system was hugely controversial, yet his successors celebrated him as a great hero of botany. PMID- 12642141 TI - A field of great promise: soil bacteriology in America, 1900-1925. AB - In the first decades of the 20th century, soil bacteriologists promised to revolutionize farming practice, much in the same way that medical bacteriologists, in the previous century, had transformed pathology, public health and sanitary engineering. Following the isolation of the microorganisms responsible for nitrification and nitrogen fixation, American soil scientists anticipated the time when farmers could 'seed' their crops and lands with these beneficial bacteria. Soil bacteriologists, during the early 20th century, never fulfilled the promise of supplying a biological source of unending soil fertility. However, in their search for productive microbes, these same researchers directed attention to the underappreciated dimensions of bacterial metabolism and microbial ecology. PMID- 12642142 TI - Jeanne Baret: the first woman to circumnavigate the globe. AB - The voyages of scientific discovery conjure in our minds images of Sir Hans Sloane bioprospecting in Jamaica in 1687 or Joseph Banks voyaging aboard the Endeavour to Tahiti and New Zealand in 1768. But women also set foot on rickety and unsure ocean-going vessels in the 18th century in the service of science. The German-born Maria Sibylla Merian voyaged from Amsterdam to Surinam in South America in 1699 to search for exotic caterpillars. She sought one that would produce a thread to rival silk, a costly and much sought-after fabric in early modern Europe. In the process, she produced one of the most celebrated 18th century natural history books. PMID- 12642143 TI - In the sign of Galileo: pictorial representation in the 17th-century Copernican debate. AB - After Galileo had discovered the four moons of Jupiter in 1609 he became increasingly convinced that the Copernican, heliocentric system of the world was correct. However, this ran against the opinions of the Church and a large number of contemporary astronomers and natural philosophers. The ensuing development culminated in the condemnation of the Copernican system by the Church in 1616 and of Galileo himself, who had propagated the Copernican system in his Dialogue Concerning the Two Chief World Systems (1632), in 1633. Nevertheless, there was a constant debate about the right world system during the whole 17th century. Pictorial representation played an important role in it and the illustrations used as book frontispieces were a significant medium for the dispute. PMID- 12642144 TI - Integrating mind and brain: Warren S. McCulloch, cerebral localization, and experimental epistemology. AB - Recently, historians have focused on Warren S. McCulloch's role in the cybernetics movement during the 1940s and 1950s, and his contributions to the development of computer science and communication theory. What has received less attention is McCulloch's early work in neurophysiology, and its relationship to his philosophical quest for an 'experimental epistemology' - a physiological theory of knowledge. McCulloch's early laboratory work during the 1930s addressed the problem of cerebral localization: localizing aspects of behaviour in the cerebral cortex of the brain. Most of this research was done with the Dutch neurophysiologist J.G. Dusser de Barenne at Yale University. The connection between McCulloch's philosophical interests and his experimental work can be expressed as a search for a physiological a priori, an integrated mechanism of sensation. PMID- 12642145 TI - The philosophy of chemistry. AB - Although chemistry is by far the largest scientific discipline according to any quantitative measure, it had, until recently, been virtually ignored by professional philosophers of science. They left both a vacuum and a one-sided picture of science tailored to physics. Since the early 1990s, the situation has changed drastically, such that philosophy of chemistry is now one of the most flourishing fields in the philosophy of science, like the philosophy of biology that emerged in the 1970s. This article narrates the development and provides a survey of the main topics and trends. PMID- 12642146 TI - Complementary and alternative medicine (CAM) in reproductive-age women: a review of randomized controlled trials. AB - PURPOSE: Complementary and alternative medicine (CAM) therapies are widely used in the general population. This paper reviews randomized controlled trials of CAM therapies for obstetrical and gynecologic conditions and presents therapies that are likely to be used by women of reproductive age and by pregnant women. DATA SOURCES: Sources included English-language papers in MEDLINE 1966-2002 and AMED (1985-2000) and the authors' extensive holdings. STUDY SELECTION: Randomized controlled clinical trials of CAM therapies for obstetric and gynecologic conditions. DATA EXTRACTION: Clinical information was extracted from the articles and summarized in tabular form or in the text. DATA SYNTHESIS: Ninety-three trials were identified, 45 of which were for pregnancy-related conditions, 33 of which were for premenstrual syndrome, and 13 of which were for dysmenorrhea. Data support the use of acupressure for nausea of pregnancy and calcium for PMS. Preliminary studies indicate a role for further research on Vitamin B6 or ginger for nausea and vomiting of pregnancy; calcium, magnesium, Vitamin B6, or chaste tree berry extract for PMS; and a low-fat diet, exercise, or fish oil supplementation for dysmenorrhea. CONCLUSIONS: Limited evidence supports the efficacy of some CAM therapies. Exposure of women of reproductive age to these therapies can be expected. PMID- 12642147 TI - Adrenal-mediated rather than direct effects of nicotine as a basis of altered sex steroid synthesis in fetal and neonatal rat. AB - Prenatal nicotine interferes with rat sexual brain differentiation and may influence human puberty. We studied effects of nicotine and cotinine on perinatal steroid synthesis in offspring of time-pregnant rats. In vitro, cotinine inhibited testosterone synthesis in neonatal rat testis. Both compounds inhibited the brain aromatase of gestational day (GD) 19 male fetuses. Effective concentrations were higher than levels in maternal plasma and fetal tissue at GD 19 after nicotine treatment from GD 12, even though nicotine accumulated in fetal brain. In view of a dual effect of nicotine in male GD 18 fetuses, decreasing plasma testosterone and increasing corticosterone [Dev Brain Res 1991;62:23-31], we administered metyrapone on GD 17 to nicotine-treated dams. 11beta-Hydroxylase inhibition completely reversed the nicotine-induced reduction of plasma testosterone at GD 18. POMC mRNA in anterior pituitary of nicotine-exposed GD 18 fetuses was reduced, probably as a result of corticosterone feedback. These data reveal a novel type of interaction of nicotine with the fetal gonadal axis involving the adrenal. PMID- 12642148 TI - Di(2-ethylhexyl) adipate (DEHA) induced developmental toxicity but not antiandrogenic effects in pre- and postnatally exposed Wistar rats. AB - Di(2-ethylhexyl) adipate (DEHA) has replaced the phthalates in thin plasticized polyvinyl chloride films used for food packaging, mainly because some phthalates induce testis toxicity and antiandrogenic effects. A dose-range finding study followed by a dose-response/effect study in Wistar rats investigated whether pre- and postnatal DEHA doses of 0, 800, or 1200mg/kg/day body weight and doses of 0, 200, 400, or 800mg/kg/day (main study) elicited developmental toxicity including antiandrogenic effects. In the main study, DEHA induced a prolonged gestation period (800mg/kg/day) and a dose-related increase in postnatal death (400 and 800mg/kg/day). DEHA also induced a permanent decrease in offspring body weight (800mg/kg/day). No antiandrogenic endpoints were affected. We conclude that DEHA induced developmental toxicity and the NOAEL is 200mg/kg. DEHA did not induce antiandrogenic effects similar to those of di(2-ethylhexyl) phthalate even though the chemical structures have similarities and the two chemicals have a common metabolite. PMID- 12642150 TI - Pharmacokinetics of D,L-3-hydroxy-3-ethyl-3-phenylpropionamide (HEPP) in pregnant rats at different stages of gestation and maternal-fetal disposition during late pregnancy. AB - The pharmacokinetics of D,L-3-hydroxy-3-ethyl-3-phenylpropionamide (HEPP), an investigational anticonvulsant drug, was evaluated in nonpregnant and in pregnant rats on gestation day (GD) 7, 12, and 21 after an intraperitoneal (i.p.) dose of 50mg/kg. Maternal-fetal disposition in the GD21 group was also evaluated. In all groups, HEPP was rapidly absorbed and the disposition was well described by an open two-compartment kinetic model. The most pronounced effects of pregnancy on the kinetics of HEPP were observed at GD21 in which significant increases in the first-order hybrid disposition rate constants alpha and beta, with corresponding decreases in half-lives were observed. Gestation also affected the intercompartmental transfer rate constants k(12) and k(21), specially at GD12 and at GD21. These changes could be associated with the physiologic increases in blood flow and cardiac output of pregnancy. There was also a slight decrease in the apparent volume of distribution at GD21, and a progressive decrease in the clearance values normalized by the body weight. No other significant differences in kinetic parameters were observed. On GD21, HEPP rapidly transfers from maternal blood to fetuses, to reach concentrations in the placenta and fetuses slightly higher than those of the maternal plasma (fetal:maternal ratio ranging from 1.07 to 1.45). After equilibrium, the concentrations in maternal, placental, and fetal tissues decreased in parallel. PMID- 12642149 TI - Folic acid awareness and intake survey in the United Arab Emirates. AB - OBJECTIVE: The aim of this study was to determine the level of knowledge and use of periconceptional folic acid supplementation in a sample of postpartum women recruited from three hospitals. DESIGN: Cross-sectional survey in which a structured questionnaire was used in a face-to-face encounter between the subject and a trained nurse. SETTINGS: Two teaching hospitals associated with Faculty of Medicine and Health Sciences and one private hospital. SUBJECTS: Postpartum women in the three hospitals were recruited during a 40-day period in November 1999. Women who did not agree to participate, had complicated labor, delivered babies with congenital malformations, or were too exhausted or difficult to examine, were excluded. RESULTS: Univariate analyses showed that overall 46.4% of the respondents had heard about folic acid and only 8.7% knew that it prevented birth defects. 45.5% of respondents took folic acid in the first trimester. The percentage of women who had ever heard about folic acid was higher in those with higher education, and those who were not UAE nationals. Use of folic acid was associated with non-UAE nationality. CONCLUSION: Awareness of the value of periconceptional folic acid was very low and use of folic acid was less prevalent among women of UAE nationality. PMID- 12642151 TI - The presence of environmental pollutants in the follicular fluid of farm animals (cattle, sheep, goats, and pigs). AB - Organochlorine pesticides and polychlorinated biphenyls are widely used in agriculture and industry, respectively. The present study assessed the burden of environmental pollutants in the follicular fluid of farm animals (cattle, sheep, goats, and pigs). An analytical method combining a solid-phase extraction with (C(18)) for clean-up and GC-electron capture detection using a capillary column was implemented for isolation and determination of organochlorine pesticides and polychlorinated biphenyls (PCBs). Of the organochlorine pesticides, hexachlorobenzene (HCB), hexachlorocyclohexane (HCH, alpha-, beta-, and gamma isomers), dieldrin, heptachlor epoxide, and the DDT-related chemicals (o,p'-DDE, p,p'-DDE, p,p'-DDD, p,p'-DDT) were detected and of the PCBs, the congeners PCB 52, -101, -138, -153, and -180 were detected. In all species of farm animals, the most frequently detected pollutant was gamma-HCH (90-100% of samples) followed by HCB (80-100%), and p,p'-DDE (75-90.91%). Species differences in the concentrations of HCB, beta-HCH, heptachlor epoxide, and DDT-related chemicals in follicular fluid were noted as well as differences in the concentrations of some pollutants within the same species. PMID- 12642152 TI - Pyridines in cigarette smoke inhibit hamster oviductal functioning in picomolar doses. AB - Past studies showed that chemicals in cigarette smoke inhibit oviductal functioning in vivo and in vitro. The purposes of this study were to identify individual toxicants in cigarette smoke solutions that inhibit various aspects of oviductal functioning and to determine their effective doses using in vitro bioassays. Solid phase extraction and gas chromatography-mass spectrometry (GC MS) were used to identify individual chemicals in mainstream (MS) and sidestream (SS) cigarette smoke solutions. Pyridines, which were the most abundant class of compounds identified, were purchased, assayed for purity, and tested in dose response studies on hamster oviducts. The lowest observable adverse effect level was determined for each pyridine derivative using the oocyte pick-up rate, ciliary beat frequency, and infundibular muscle contraction assays. 2 Methylpyridine, 4-methylpyridine, 2-ethylpyridine, 3-ethylpyridine, and 4 vinylpyridine were inhibitory at picomolar concentrations in all assays. This work shows picomolar doses of pyridines with single methyl or ethyl substitutions significantly inhibit oviductal functioning raising questions regarding the safety of these compounds. PMID- 12642153 TI - Developmental toxicity study of chlorpyrifos in rats. AB - Chlorpyrifos (O,O-diethyl-O-(3,5,6-trichloro-2-pyridyl)-phosphorothioate) was evaluated for potential developmental toxicity. Groups of 30 bred female Fischer 344 rats were given 0, 5, 15, and 25mg/kg per day by gavage on gestation days 6 15; the fetuses were evaluated on gestation day 21. Clinical signs of toxicity attributed to chlorpyrifos were noted in dams receiving 15 and 25mg/kg per day. Maternal effects in these groups also included depressed body weight and acetylcholinesterase activity. Fetal weight and viability were decreased, and fetal death and early resorption were increased at the 25mg/kg per day maternal dose. Visceral, skeletal, and external variations were also increased in this group. Chlorpyrifos showed fetotoxic and teratogenic effects at a maternal dose of 25mg/kg per day, a dose that also produced maternal toxicity. PMID- 12642154 TI - Influence of the mycotoxins alpha- and beta-zearalenol and deoxynivalenol on the cell cycle of cultured porcine endometrial cells. AB - The present study investigated the effects of the mycotoxins alpha-zearalenol (alpha-ZOL) and beta-zearalenol (beta-ZOL) at concentrations of 7.5, 15, and 30 microM, and deoxynivalenol (DON) at concentrations of 0.94, 1.88, and 3.76 microM on cell cycle distribution (propidium iodide, PI staining) in combination with the proliferating cell nuclear antigen (PCNA) by flow cytometry. The viability of porcine uterine cells was not impaired at 30 microM alpha-ZOL, whereas beta-ZOL at this concentration and 3.76 microM DON significantly decreased cell number. Some cells showed ultrastructural features of cell death indicated by swollen mitochondria, disrupted cell membranes, and many vacuoles. After 24 and 48h of exposure to alpha-ZOL (7.5, 15, or 30 microM), the cell cycle distribution was still comparable to the control groups. An anti-proliferative effect of beta-ZOL and DON was detected by a significant reduction in the S-phase together with arrest of cells in the G(0)/G(1)-phase. The results show that beta-ZOL (7.5, 15, or 30 microM) and DON (0.94, 1.88, or 3.76 microM) control the progression of cells through the cycle by decreasing S-phase and arresting cells in the G(0)/G(1)-phase of the cell cycle. A significant decrease in the expression of the proliferation marker PCNA amounts indicates that beta-ZOL and DON disengaged cells from active cycling. We confirm that alpha-ZOL possesses a relative binding affinity to porcine uterine cytoplasmic estrogen receptor. PMID- 12642155 TI - Metallothionein gene expression under different time in testicular Sertoli and spermatogenic cells of rats treated with cadmium. AB - The rodent testes are generally more susceptible to cadmium (Cd)-induced toxicity than the liver. To clarify the molecular mechanism underlying tissue and cell differences in Cd sensitivity, we compared metallothionein (MT) gene expression, MT protein accumulation, and Cd retention under different times in freshly isolated testicular Sertoli and spermatogenic cells and liver of rats treated with Cd. Adult male Sprague-Dawley rats received a s.c. injection of 4.0 micromol Cd/kg and 1, 3, 6, or 24h later and untreated animals (0h) tissue were sampled and testicular Sertoli and spermatogenic cells isolated. MT1 and MT2 mRNA levels were determined by semi-quantitative RT-PCR analysis followed by densitometry scanning, and MT was estimated by the enzyme-linked immunosorbent assay (ELISA) method. Cadmium content was determined by atomic absorption spectrophotometry. Testicular lesions were not grossly or histologically observed in rats treated with 4.0 micromol Cd/kg. In the present study, we demonstrated that the rat testis indeed expressed MT1 and MT2, the major isoforms. We also found that untreated animals contained relatively high basal levels of both isoform mRNA, which were increased after Cd treatment in liver and peaked at 3h, followed by a decline, in contrast, the mRNA levels in Sertoli cells peaked at 6h. Interestingly, the induction of MT1 mRNA was lower than MT2 mRNA in Sertoli cells and liver of rats treated with Cd. However, the MT1 mRNA levels of spermatogenic cells decreased 0-3h after Cd treatment, followed by an increase; in contrast, MT2 mRNA levels increased 0-3h after Cd treatment, followed by a reduction, but induced extents of them are lower than those of Sertoli cells and liver. Cd exposure substantially increased hepatic MT, but did not increase MT translation in Sertoli and spermatogenic cells. These results indicate: (1) that Cd-induced MT mRNA expression is cell- and time-dependent; (2) that the inability to induce the metal-detoxicating MT protein in response to Cd, might account for higher susceptibility of testes to Cd toxicity and carcinogenesis relative to liver. PMID- 12642156 TI - Effects of single non-ortho, mono-ortho, and di-ortho chlorinated biphenyls on cell functions and proliferation of the human prostatic carcinoma cell line, LNCaP. AB - This study investigated the effects of the non-ortho 3,3',4,4',5 pentachlorobiphenyl (PCB 126) and 3,3',4,4'-tetrachlorobiphenyl (PCB 77), the mono-ortho 2,3',4,4',5-pentachlorobiphenyl (PCB 118), and the di-ortho substituted 2,2',4,4',5,5'-hexachlorobiphenyl (PCB 153) on the human prostatic carcinoma cell line LNCaP. Activities of 5alpha-reductase and ethoxyresorufin-O deethylase (EROD) secretion of testosterone-regulated prostatic specific antigen (PSA) and cell proliferation served as specific and general cell markers. The two non-ortho (dioxin-like) PCBs, PCB 126 and PCB 77, showed anti-androgenic properties. Both PCBs reduced androgen-depending PSA secretion and cell proliferation, and inhibited the DHT-producing enzyme 5alpha-reductase in a concentration-dependent manner. In contrast, the ortho-substituted PCBs, PCB 118 and PCB 153, had no effect on 5alpha-reductase. However, they had a biphasic effect on LNCaP cell proliferation. PCB 153 and, to some extent, PCB 118 induced cell proliferation and PSA secretion at low concentrations, whereas, these parameters were reduced at high concentrations. Since EROD induction and inhibition of 5alpha-reductase activity was not observed, these findings suggest an AhR and AR independent mechanism and post-transcriptional target sites should be considered. The effects of PCB 126 on cell proliferation, PSA secretion, and 5alpha-reductase activity may be partly a result of AhR-dependent inhibition. In conclusion, the endocrine activity of PCBs via direct steroid hormone receptor mediated and Ah receptor-mediated pathways are well known. However, the modulation of post-transcriptional targets may account for some of the endocrine disrupting properties of single PCB congeners. PMID- 12642158 TI - Effects of dinoseb, 4,6-dinitro-o-cresol, and 2,4-dinitrophenol on rat Sertoli germ cell co-cultures. AB - The effects of dinoseb (DNBP), a known testicular toxicant in the rat, on germ cells were investigated in Sertoli-germ cell co-cultures. Two DNBP-related dinitrophenolic compounds, 4,6-dinitro-o-cresol (DNOC) and 2,4-dinitrophenol (DNP), were also examined, as testicular toxicity of these compounds had not been elucidated. Cultures were exposed to each compound (10(-7)-10(-4)M) for 24h and examined for the number and viability of detached cells and morphologic alterations under a light microscope. DNBP significantly increased the number of detached cells (10(-5) and 10(-4)M) and suppressed their viability (10(-6)-10( 4)M). Morphologic observations revealed degenerative alterations in the germ cells and Sertoli cells. Similar effects as observed after DNBP exposure were evident at 10(-4)M DNOC and 10(-4)M DNP. These results demonstrate that DNBP, DNOC, and DNP have in vitro toxicity to these cell populations at high concentration, and suggest the possibility that DNOC and DNP also cause testicular damage in experimental animals and humans. PMID- 12642157 TI - Effects of single dose administration of ifosfamide on testes and semen characteristics in the rabbit. AB - The effects on testes and semen characteristics were studied in the rabbit after a single dose administration of the chemotherapeutic agent ifosfamide. Sexually mature male rabbits received a single intravenous injection of either 0, 60, 90, 120, or 240mg/kg body weight ifosfamide. Two phases of experimentation were conducted, lasting 1 and 18 weeks, respectively, at the end of which half of the animals from each treatment group were euthanized. Reproductive organs weighing, as well as testicular qualitative and quantitative histological examinations were performed 1 and 18 weeks post-treatment, while semen quality and libido were evaluated on a weekly basis. A decrease in the paired testes weight (90, 120, and 240mg/kg groups) and the accessory sex glands weight (240mg/kg group) were noted 1 week post-treatment. Although no histopathologic lesions or significant changes in the quantitative histologic examination were observed, semen quality examination revealed transient oligospermia (60, 90, 120, and 240mg/kg groups), teratozoospermia (120 and 240mg/kg groups), and asthenozoospermia (240mg/kg group), which returned to normal by the 6th (60 and 90mg/kg groups), 7th (120mg/kg group), or 8th week after treatment (240mg/kg group). Libido remained normal. The results suggest that ifosfamide, at a single dose, causes transient and dose-dependent depression of spermatocytogenesis (240mg/kg), spermiogenesis (60, 90, and 120mg/kg), and sperm maturation in the epididymis (240mg/kg). PMID- 12642159 TI - Limit dose testing, lets not forget animal welfare. PMID- 12642160 TI - Short- and long-term influence of perinatal dexamethasone treatment on swine growth. AB - The objective was to evaluate the effects of perinatal dexamethasone (Dex) treatment on postnatal growth in pigs. Experiment 1: 42 piglets were assigned according to birth weight and sex to receive either Dex (1 mg/kg body weight) or sterile saline (Control; equivalent volume) i.m. within 1h of birth. Body weights were recorded weekly and at sacrifice (day 18). Birth weights (1.43 +/- 0.05 kg) did not differ between treatment groups (P > 0.19). At day 18, Dex pigs were heavier than Control pigs (5.46 +/- 0.24 and 4.45 +/- 0.26 kg, respectively). Serum IGF-1 was 17.3% higher in Dex pigs (P < 0.04) compared to Controls. For serum GH, there was a treatment x sex interaction (P < 0.04) with GH being 51% lower in Dex males compared to Control males, and no differences in females. Experiment 2: 71 pigs were assigned according to birth weight and sex to receive either Dex (2 mg/kg body weight) or sterile saline (Control; equivalent volume) i.m. within 1 h of birth. Body weights were recorded weekly until weaning (day 21) and then every 14th day until market weight. Birth weights (1.53 +/- 0.03 kg) did not differ (P > 0.35) between treatment groups or sexes. Dexamethasone increased growth from birth to market weight by 4.15%. Carcass weights were not different (P > 0.34) between Dex (89.9 +/- 1.17 kg) and Control pigs (88.6 +/- 1.36 kg). Overall, Dex enhanced growth in pigs from birth to market weight with minimal effects on carcass and meat quality. PMID- 12642161 TI - Effects of Ghrelin on growth hormone secretion from cultured adenohypophysial cells in pigs. AB - To clarify the direct effects of Ghrelin on growth hormone (GH) release from anterior pituitary (AP) cells in pigs, GH-releasing effects of human Ghrelin (hGhrelin) and rat Ghrelin (rGhrelin) on porcine AP cells were compared with GHRH in vitro. The AP cells were obtained from 6-month-old pigs and the cells (2 x 10(5) cells per well) were incubated for 2 h with the peptides after incubating in DMEM for 3 days. hGhrelin and rGhrelin significantly stimulated GH release from the cultured cells at doses of 10(-8) and 10(-7)M (P < 0.05). The rates of increase in GH at 10(-8) and 10(-7)M of hGhrelin were 82.7 and 131.9%, while those with rGhrelin were 43.9 and 79.5%, respectively. GHRH significantly stimulated GH release from the cells at a dose as low as 10(-11)M (P < 0.05), and the response to GHRH was greater than that induced by Ghrelins. In time-course experiments, GHRH continued to increase GH concentrations in media until 120 min after incubation; however, those in media treated with hGhrelin reached a plateau 60 min after incubation, and the maximal value was approximately one third that obtained with GHRH. When hGhrelin (10(-8)M) and GHRH (10(-8)M) were added together, additive effects of both peptides on the release of GH were observed (P < 0.05). Somatostatin (SS, 10(-7)M) significantly blunted GH release induced by hGhrelin (10(-8)M) and GHRH (10(-8)M) (P < 0.05). In the presence of SS, additive effects of hGhrelin and GHRH on the release of GH were observed (P < 0.05). These results show that Ghrelin directly stimulates GH release from anterior pituitary cells in pigs; however, the GH-releasing effect is weaker than that of GHRH in vitro. The present results also show that Ghrelin interacts with GHRH and SS to in the release of GH from porcine adenohypophysial cells. PMID- 12642162 TI - Relationships between plasma concentrations of leptin and other metabolic hormones in GH-transgenic sheep infused with glucose. AB - To study the regulation of leptin secretion in sheep, we infused glucose (0.32 g/h/kg for 12 h) into GH-transgenic animals (n = 8) that have chronically high plasma concentrations of ovine GH and insulin, but low body condition and low plasma leptin concentrations, and compared the responses with those in controls (n = 8). In both groups, the infusion increased plasma concentrations of glucose and insulin within 1 h and maintained high levels throughout the infusion period (P < 0.0001). Compared with controls, GH-transgenics had higher concentrations of insulin, IGF-1, GH (all P < 0.0001) and cortisol (P < 0.05), but lower GH pulse frequency (P < 0.0001). Overall, leptin concentrations were lower in GH transgenics than in controls (P < 0.01). A postprandial increase in leptin concentrations was observed in both groups, independently of glucose treatment, after which the values remained elevated in animals infused with glucose, but returned to basal levels in those infused with saline, independently of transgene status. In both GH-transgenics and controls, glucose infusion did not affect the concentrations of GH, IGF-1, or cortisol. In conclusion, GH-transgenic and control sheep show similar responses to glucose infusion for leptin and other metabolic hormones, despite differences between them in body condition and basal levels of these hormones. Glucose, insulin, GH, IGF-1 and cortisol are probably not major factors in the acute control of leptin secretion in sheep, although sustained high concentrations of GH and IGF-1 might reduce adipose tissue mass or inhibit leptin gene expression. PMID- 12642163 TI - Methimazole and thyroid hormone replacement in broilers. AB - Seven-day-old chickens were fed diets containing 18% crude protein + 0 or 1g methimazole/kg to produce either euthyroid or hypothyroid groups of birds at 28 days of age. These two groups were then offered diets containing either 0 or 1mg triiodothyronine (T(3))/kg diet. Birds were sampled at 0, 2, 5, and 8 days following the onset of the T(3) treatment. Measurements taken at these intervals included in vitro hepatic lipogenesis (IVL), growth and feed consumption, hepatic enzyme activities (malic enzyme, ME; isocitrate dehydrogenase, ICD; and aspartate amino transferase, AAT), plasma hormones (T(3); thyroxine, T(4); insulin like growth factors I, IGF-I; and insulin like growth factors II, IGF-II) and metabolites (glucose; fatty acids, NEFA; triglyerides; uric acid). Hypothyroidism decreased IVL and ME at 28 days of age; however, T(3) supplementation for 2 days restored both IVL and ME. Paradoxically, continuing T(3) replenishment for an additional 3-6 days decreased IVL without affecting ME activity. In contrast, supplemental T(3) decreased IVL in euthyroid birds, regardless of the dosing interval, but had no effect on ME activity. Methimazole decreased plasma T(3), T(4), uric acid, and IGF-I, but did not affect IGF-II at 28 days. Giving T(3) to birds previously on methimazole increased plasma IGF-I as did feeding a control diet. Supplemental T(3) increased NEFA in both euthyroid and hypothyroid birds, but only for a short period following the initiation of supplementation (2 days post-supplementation). These data may help to explain some of the apparent reported dichotomies in lipid metabolism elicited by changes in the thyroid state of animals. In addition, most metabolic changes in response to feeding T(3) occurred within 2-5 days, suggesting that changes in intermediary metabolism preceded morphological changes. In conclusion, the thyroid state of the animal will determine responses to exogenous T(3). PMID- 12642164 TI - Body composition, leptin, and the leptin receptor and their relationship to the growth hormone (GH) axis in growing wethers treated with zeranol. AB - Age-related changes in body composition, leptin, and hypothalamic-pituitary expression of the leptin receptor and associative relationships of these factors to constituents of the growth hormone (GH) axis were evaluated. Seventy wethers were randomly assigned at birth to one of four treatment groups: control; treatment 1 implanted with the estrogenic compound zeranol (12 mg, Ralgro on days 0, 45, and 90; treatment 2 received zeranol on days 45 and 90; and treatment 3 received zeranol on day 90. Serum and tissues were collected from wethers (n > or = 5) from each group on days 28, 73, 118, and 135. Percent body fat and leptin increased linearly (P < 0.01) with age, but were not influenced (P > or = 0.14) by zeranol. The leptin receptor in the pituitary appeared to be differentially (P = 0.097) expressed across days 73-135, but no differences (P > or = 0.43) were detected in expression of this receptor in the hypothalamus among treatments and ages. Leptin and % body fat were negatively correlated (r > or = -0.52, P < 0.05) to mRNA levels of factors involved in pituitary synthesis and secretion of GH. Serum leptin increased with age as did percent body fat, but zeranol did not influence body composition, serum leptin, or expression of the leptin receptor in the hypothalamus or pituitary; however, the leptin receptor appeared to be differentially expressed among the hypothalamus and pituitary with level of body fat and leptin being inversely associated to transcriptional-factors involved in somatotrope synthesis and secretion of GH. PMID- 12642166 TI - Female prairie voles do not choose males based on their frequency of scent marking. AB - We conducted an experiment to test three alternative hypotheses for the function of frequency of scent marking in male prairie voles, MICROTUS OCHROGASTER: (1) sexual attraction (to advertise male quality for mating); (2) reproductive competition; and (3) self-advertisement or individual identity. In laboratory experiments, males deposited scent on all areas of a bare substrate, and more in an area next to a stimulus animal than other areas, regardless of the stimulus animal's sex. Females did not choose mates based on their frequency of scent marking and scent marking did not antagonize or stimulate aggression between males. The frequency of scent marking by males supports the individual identity hypothesis, and is less consistent with the sexual attraction or reproductive competition hypotheses. Mate choice is likely based on a complex suite of characters, but at least in prairie voles, the frequency of scent marking by males does not appear to be one of them. PMID- 12642165 TI - Partial nucleotide sequence of the porcine corticosteroid-binding globulin (CBG) cDNA and specification of CBG expression sites in postnatal pigs. AB - The purpose of this study was to develop a porcine CBG cDNA probe in order to examine the porcine CBG mRNA expression in major tissues from the postnatal pig. The reverse transcriptase-polymerase chain reaction (RT-PCR) was conducted to develop the porcine CBG cDNA probe using total RNA extracted from liver of 40-day old pig. The RT-PCR product was subcloned into the pGEM vector (Promega, Madison, WI) and subjected to restriction enzyme treatments and DNA sequencing. Northern blot analysis was conducted using total RNA extracted from samples (approximately 200 mg) of liver, lung, kidney, and whole adrenal tissue that were collected from pigs on day 3 (n = 2) or day 40 (n = 2) postpartum. A 500 bp partial porcine CBG cDNA encoded 166 amino acids and had 83, 78, and 77% homology to a 494-nucleotide sequence of CBG from sheep, human, and rabbit, respectively. The deduced peptide sequence of the partial porcine CBG showed 77, 62, 60, and 51% homology to sheep, human, rabbit, and rat CBG sequences, respectively. An approximately 1.53 kb CBG mRNA was detected only in the liver tissue. In conclusion, the development of a partial CBG cDNA for swine makes it possible to study the ontogeny and the regulation of CBG synthesis at the molecular level and, based on tissues examined in this study, the liver appears to be the primary source of CBG biosynthesis in the postnatal pig. PMID- 12642167 TI - Increased effort requirements and risk sensitivity: a comparison of delay and magnitude manipulations. AB - Reward magnitude and delay to reward were independently manipulated in two separate experiments examining risk-sensitive choice in rats. A dual-running wheel apparatus was used and the tangential force resistance required to displace both wheels was low (50g) for half of the subjects, and high (120g) for the remaining subjects. Concurrent FI30-s and FI60-s schedules delivered equivalent amounts of food reward per unit time (i.e. 5 and 10 pellets of food, respectively), and these conditions served as the baseline treatment for all subjects. Variability, either in reward magnitude or delay, was introduced on the long-delay (60s) schedule during the second phase. All subjects were returned to the baseline condition in the third phase, and variability was introduced on the short-delay (30s) interval schedule during phase four. The subjects were again returned to the baseline condition in the fifth and final phase, ultimately yielding a five-phase ABACA design. Original baseline performance was characterized by a slight short-delay interval preference, and this pattern of performance was recovered with each subsequent presentation of the baseline condition. Overall, the data obtained from the reward magnitude and delay-to reward manipulations were indistinguishable; subjects experiencing low-response effort requirement behaved in a risk-indifferent manner and subjects experiencing high-response effort requirement preferred the variable schedule. Implications for the daily energy budget rule on risk-sensitive foraging are discussed in light of these findings. PMID- 12642168 TI - Behavioural responses of farm mink towards familiar and novel food. AB - The study aimed to (i) describe the response of farmed mink towards familiar and novel food, and (ii) assess the suitability of using novel food in order to measure fear responses in mink. A total of 48 farm mink from two behaviourally selected genetic lines were caged individually with one standardised daily feeding. The experimental feedings were carried out in a balanced order on 2 successive days. Behaviour of the mink was video recorded for 10min after food provision. In general, the mink were reluctant to approach and eat the novel food compared to familiar food, which indicates a neophobic feeding strategy, i.e. increased latency to come close to (P<0.01) and eat (P<0.001) the novel food, and a higher frequency (P<0.001) and a longer duration (P<0.001) of sniffing. Provision of unknown food items may be used to evaluate fear responses in mink since mink respond reluctantly towards novel food. However, high inter-individual variation in the present study obliterated a previously shown difference between confident and fearful mink, suggesting that this method for measuring fear responses in mink is appropriate only on a large animal material. PMID- 12642169 TI - Variability of vibratory signals and mate choice selectivity in the southern green stink bug. AB - The southern green stink bug, Nezara viridula, has a complex mate recognition system that implicates chemical and acoustic signals. To localise a sexual partner acoustically, a male and female alternate between the male courtship song (MCrS) and the female calling song type 1 (FCS1). Although previous research has revealed that both signals show geographical variability, until now no studies have explored the form of this variability. We analysed the temporal and spectral characteristics of MCrS and FCS1 pulse trains of males and females from a French and a Guadeloupe population. Pulse train duration of the MCrS varied within and between populations. Likewise, spectral and temporal parameters of FCS1 varied within and between the two populations. Although females did not show any behavioural responses to pre-recorded MCrS, males responded to pre-recorded FCS1 by emitting a higher number of MCrS per minute. Furthermore, males modulated the repetition rate of their MCrS pulse trains to match those of the FCS1. All males responded to FCS1 from French and Guadeloupe females despite the temporal differences in these songs; however, they responded with a shorter latency and a higher rate of MCrS/FCS1 pulse trains to the songs of females from their own population. In choice experiments with two alternatives, responses to FCS1 were inhibited when males received a simultaneous female call from another Pentatomidae sympatric species, Acrosternum hilare. We conclude that, although males prefer FCS1 from their own population, they recognise FCS1 from French and Guadeloupe N. viridula females as species-specific female calls. Variability of vibratory signals might play a role in mate choice. PMID- 12642170 TI - Ontogeny has a phylogeny: background to adjunctive behaviors in pigeons and budgerigars. AB - Animals coping with operant conditioning tasks often show behaviors that are not recorded by keys, levers and similar response transducers. Nevertheless, these adjunctive behaviors should not be disposed of by classifying them as incidental. Often they are found to be at least partially influenced by the experimentally programmed contingencies, and under certain conditions they can in turn influence conditioned behaviors. Here we describe the occurrence and characteristics of two such behaviors, stimulus grasping in operantly key-pecking pigeons and intra delay stereotypies in a delayed matching-to-sample task with budgerigars. It is argued that for a proper account of these behaviors it is necessary to refer to a behavioral systems approach that appeals to longer ranging ontogenetic and phylogenetic histories than is usually considered in the psychological literature. The gaping towards on-key stimuli by pigeons is attributed to the hypothesis that operantly conditioned key-pecks probably relate to a grasp pecking response that is normally executed towards non-edible items covering food. The intra-delay behaviors shown by the budgerigars are assumed to have originated from stress-induced displacement responses that adventitiously came under the influence of differential reinforcement contingencies. Finally, we discuss what kinds of evidence are needed to put these hypothetical explanations on a more certain footing. PMID- 12642171 TI - Multiple exposure to activity anorexia in rats: effects on eating, weight loss, and wheel running. AB - Animals were given five cycles of an activity anorexia (AA) procedure in order to determine the effect of additional experience on eating, running, and weight loss. Female Sprague-Dawley rats were given a 1h meal and allowed access to a running wheel for the remainder of each day. Upon reaching 75% of free-feeding body weight, each animal was denied wheel access and given ad libitum food until it regained the lost weight. Then, food was again restricted and wheel access provided. Sedentary control animals were placed on the restricted feeding schedule for the median number of days experimental animals required to reach weight loss criterion. Experimental animals showed adaptation by increasing food consumption and decreasing the rate of weight loss despite an increase in running across cycles. Additionally, the distribution of running shifted gradually so that during the later cycles, much of the running occurred in the hours just before feeding. The results support the hypothesis that running interferes with adaptation to the restricted feeding schedule and also that the marked increase in anticipatory behavior during the later cycles is primarily responsible for the maintenance of AA. PMID- 12642172 TI - Toward a molecular architecture of personality. AB - Epidemiological studies provided a large body of evidence that personality dimensions are influenced by genetic factors and that the genetic component is highly complex, polygenic, and epistatic. However, consistent findings on the genetic basis of personality have yet remained sparse. In recent years, molecular genetics has begun to identify specific genes coding in particular for components of the serotonergic and dopaminergic neurotransmitter systems representing quantitative trait loci (QTLs) for behavioral traits. The QTL concept suggests that complex traits are not attributable to single genes. According to this polygenic model, the genetic basis of personality and behavior and its pathological variations thus results from additive or nonadditive interactions of various genes. As the number of suitable candidate genes constantly increases, the QTL model provides a reasonable explanation for the genetic basis of personality and its disorders. In this review, the current knowledge on the impact of a large number of candidate gene polymorphisms (e.g. variations in serotonin and dopamine receptor and serotonin transporter genes) on personality and temperament is summarized. Additionally, investigations of gene-gene and gene environment interactions in humans and animals, which currently intensify the identification of genes that underlie behavioral variations, are examined. The findings converge on the notion that a probabilistic rather than deterministic impact of genes on the expression of behavior will contribute to the demystification of behavioral disorders. PMID- 12642173 TI - Influence of synaptic serotonin level on [18F]altanserin binding to 5HT2 receptors in man. AB - The feasibility of in vivo serotonin 5HT(2) receptor binding measurement using [18F]altanserin as a radioligand has been well established. In this study, the postsynaptic receptor binding potential of this ligand was examined as a possible indicator of synaptic serotonin content after pharmacological challenge. Studies were performed in 11 subjects with a history of recurrent major depression. Six of them received serotonergic antidepressive treatment at the time of the experiment, the other five patients were untreated. Two PET measurements were carried out in each subject within 2 or 3 days. Before one of the measurements, 25 mg of the serotonin re-uptake inhibitor clomipramine were given intravenously, the other measurement was done without pharmacological challenge. The data were analyzed using non-linear least-square regression and Logan's graphical method. In the whole group of subjects, binding potential and distribution volume of altanserin decreased following clomipramine challenge. The decrease was between 14 (P=0.03) and 23% (P=0.004). This effect was mainly seen in subjects not on antidepressive medication. Clomipramine challenge probably increased the synaptic serotonin level, which competed with altanserin leading to the lowered binding potential. The paradigm might, thus, be useful to estimate serotonin release in vivo. Pretreatment with serotonergic antidepressants reduces the effect of clomipramine. PMID- 12642174 TI - Effect of postnatal iron administration on MPTP-induced behavioral deficits and neurotoxicity: behavioral enhancement by L-Dopa-MK-801 co-administration. AB - Two experiments were performed to investigate the interactive effects of postnatal iron administration and adult MPTP treatment upon the function of C57 Bl/6 mice tested at adult age and to ascertain the possible ameliatory effects of a subthreshold dose of L-Dopa co-administered with different doses of the uncompetitive glutamate antagonist, MK-801. Experiment I indicated that postnatal iron induced marked deficits (hypoactivity), initially, in all three parameters of motor activity at the 5.0 and 7.5 mg/kg doses, and to a lesser extent at the 2.5 mg/kg dose. Later combination with MPTP (2x40 mg/kg) potentiated severely these deficits. During the final period of testing a marked hyperactivity was obtained for the two higher dose groups; this effect was abolished in mice administered MPTP. Experiment II indicated that the deficits in motor activity parameters induced by postnatal iron at 7.5 mg/kg were alleviated in a dose related manner by the co-administration of the uncompetitive glutamate antagonist, MK-801, with a subthreshold dose of L-Dopa. Postnatal iron (7.5 mg/kg) administration followed by low doses of MPTP (2x20 mg/kg) 3 months later virtually abolished all motor activity. The combination of these compounds increased also the motor activity of mice treated with MPTP (2x20 mg/kg) or mice treated with the combination of postnatal iron and MPTP. The combination of MK 801 with L-Dopa increased locomotor (0.3 mg/kg), rearing (0.1 and 0.3 mg/kg) and total activity (0.3 mg/kg) of iron-treated mice during the initial, hypoactive 30 min period of testing. Locomotor activity (0.1 mg/kg) of MPTP-treated mice was increased too during this period. During the final 30-min period of testing all three parameters of activity (locomotion, 0.3 mg/kg; rearing and total activity, 0.1 and 0.3 mg/kg) were enhanced in the iron-treated mice, locomotion (0.1 mg/kg) and rearing (0.1 mg/kg) in the iron plus MPTP treated mice and only locomotion (0.1 mg/kg) in the MPTP-treated mice. In control mice (vehicle+saline), the higher doses of MK-801 (0.1 and 0.3 mg/kg) enhanced both locomotor and total activity. Analyses of total iron concentration in the frontal cortex and basal ganglia of Fe(2+) and vehicle treated mice indicated that marked elevations basal ganglia iron levels of the 5.0 and 7.5 mg/kg groups, later injected either saline or MPTP, were obtained (Experiment I). In Experiment II, iron concentrations in the basal ganglia were elevated in both the Fe(2+)-sal and Fe(2+)-MPTP groups to 170 and 177% of Veh.-sal values, respectively. There was a significant increase in the frontal cortex of iron-treated mice later administered either saline or MPTP (2x40 mg/kg) in Experiment I as well as in those given iron followed by MPTP (2x20mg/kg) in Experiment II. The implications of iron overload in parkinsonism seem confirmed by the interactive effects of postnatal administration of the metal followed by adult MPTP treatment upon motor activity and the activity enhancing effects of co-administration of L-Dopa with MK-801. PMID- 12642175 TI - Frontal cortex BDNF levels correlate with working memory in an animal model of Down syndrome. AB - Individuals with Down syndrome (DS) develop most neuropathological hallmarks of Alzheimer's disease early in life, including loss of cholinergic markers in the basal forebrain. Ts65Dn mice, an animal model of DS, perform poorly on tasks requiring spatial memory and also exhibit basal forebrain pathology beginning around 6 months of age. We evaluated memory as well as brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) protein levels in basal forebrain, frontal cortex, hippocampus, and striatum in Ts65Dn mice at the age when cholinergic degeneration is first observed, and compared values to normosomic controls. Six-month-old Ts65Dn mice exhibited impairments in working and reference memory as assessed on a water radial-arm maze. The working memory deficit was related to the inability of Ts65Dn mice to successfully sustain performance as the working memory load increased. Coupled with cognitive performance deficiencies, Ts65Dn mice also exhibited lower frontal cortex BDNF protein levels than controls. Further, BDNF levels were negatively correlated with working memory errors during the latter portion of testing in Ts65Dn mice, thereby suggesting that lower BDNF protein levels in the frontal cortex may be associated with the observed working memory impairment. PMID- 12642176 TI - Estradiol valerate and alcohol intake: a comparison between Wistar and Lewis rats and the putative role of endorphins. AB - Studies show that estrogens can influence alcohol consumption; however, findings are variable and an etiology remains unknown. Furthermore, estrogen administration can alter several neurotransmitter systems implicated in alcohol consumption, including the beta-endorphin (beta-EP) system. The present studies investigate (a) whether estradiol valerate (EV) alters voluntary alcohol consumption in Wistar and Lewis rats, (b) if an effect of EV on drinking is associated with changes in hypothalamic or pituitary beta-EP content, and (c) whether differences in alcohol drinking between treatment and rat groups are related to locomotor or defensive behavior/anxiety scores. Of 30 Wistar and 30 Lewis rats used in this study, half were injected with 2 mg EV in 0.2 ml sesame oil, while the remainder were injected with the vehicle only. After 8 weeks, all animals were tested in the open field and elevated plus maze. A week later, 4-6 animals in each group were sacrificed. The remaining animals were tested for voluntary alcohol drinking for 24 days prior to being sacrificed on the last day. Radioimmunoassay was used to estimate hypothalamic and pituitary beta-EP content. Wistar and Lewis rats injected with EV showed an increase in alcohol drinking, but their behavior scores and beta-EP levels remained unaltered. This result suggests that any EV effect on drinking is unrelated to changes in beta-EP or behavioral performance. Furthermore, Wistar rats show higher alcohol drinking, locomotor and defensive behavior scores, and hypothalamic beta-EP than Lewis rats. Higher alcohol drinking by Wistar rats might be due to higher behavioral scores or endogenous opioid activity/sensitivity. PMID- 12642177 TI - The effects of anisomycin (a protein synthesis inhibitor) on spatial learning and memory in CA1 region of rats hippocampus. AB - Inhibition of protein synthesis has been shown to affect long-term memory in a wide variety of animal species. But little is known regarding the neuroanatomical location of protein synthesis in different memory tasks. In this study, the effect of intrahippocampal injection of anisomycin, an inhibitor of brain protein synthesis on spatial memory was examined in Morris Water Maze. At first, rats were connulated bilaterally into the CA1 region and then different doses of anisomycin (1.25-2.5 micro g/0.5 micro l) on its vehicle (saline) were injected bilaterally into the CA1 region of rats hippocampus 20 min before training each day. The results showed dose-dependent increases in latencies to find the invisible platform and traveled distances in anisomycin received group compared to the control group. Therefore, it appears that protein synthesis inhibition in the CA1 region of hippocampus impair spatial learning in Morris Water Maze. PMID- 12642178 TI - Repeated predictable or unpredictable stress: effects on cocaine-induced locomotion and cyclic AMP-dependent protein kinase activity. AB - Stressful experiences appear to have a strong influence on susceptibility to drug taking behavior. Cross-sensitization between stress and drug-induced locomotor response has been found. Locomotor response to novelty or cocaine (10 mg/kg, i.p.), cyclic AMP-dependent protein kinase (PKA) activity in the nucleus accumbens and basal corticosterone levels were evaluated in male adult rats exposed to acute and chronic predictable or unpredictable stress. Rats exposed to a 14-day predictable stress showed increased locomotor response to novelty and to cocaine, whereas rats exposed to chronic unpredictable stress demonstrated increased cyclic AMP-dependent PKA activity in the nucleus accumbens. Both predictable and unpredictable stress increased basal corticosterone plasma levels. These experiments demonstrated that stress-induced early cocaine sensitization depends on the stress regime and is apparently dissociated from stress-induced changes in cyclic AMP-dependent PKA activity and corticosterone levels. PMID- 12642179 TI - Gaze during visually-guided locomotion in cats. AB - Visual guidance is often critical during locomotion. To understand how the visual system performs this function it is necessary to know what pattern of retinal image motion neurons experience. If a locomoting observer maintains an angle of gaze that is constant relative to his body, retinal image motion will resemble Gibson's (The Perception of the Visual World (1950)) well-known optic flow field. However, if a moving observer fixates and tracks a stationary feature of the environment, or shifts his gaze, retinal motion will be quite different. We have investigated gaze in cats during visually-guided locomotion. Because cats generally maintain their eyes centered in the orbits, their gaze can be monitored with reasonable accuracy by monitoring head position. Using a digital videocamera, we recorded head position in cats as they walked down a cluttered alley. For much of the time, cats maintained a downward angle of gaze that was constant relative to their body coordinates; these episodes averaged 240 ms in duration and occupied 48-71% of the total trial time. Constant gaze episodes were separated by gaze shifts, which often coincided with blinks. Only rarely did we observe instances when cats appeared to fixate and track stationary features of the alley. We hypothesize that walking cats acquire visual information primarily during episodes of constant gaze, when retinal image motion resembles Gibson's conventional optic flow field. PMID- 12642180 TI - Behavioural changes induced by early and long-term gravito-inertial force modification in the rat. AB - The study concerns rats conceived, born and raised in a hypergravity environment (HG: 2 g) for 3 months using a centrifuge. They were then exposed to terrestrial gravity (1 g) and submitted to behavioural tests investigating their spontaneous locomotor activity (open-field), their posture (support surface), and their vestibular function (air-righting reflex). Performances were compared to age matched control rats housed at 1 g for the same time period. Results showed static and dynamic behavioural deficits as early as the rats were exposed to normal gravity. They exhibited strongly increased motor activity in open-field, with longer travelled distances and more scattered trajectories; in addition, the HG rats displayed more numerous rearings than controls did. They showed postural changes characterized by an enlarged support surface and they did not succeed in the air-righting reflex, due to increased time-delay for head righting. None of these changes were permanent. Indeed, for all tests, the HG rats tested after 3 weeks spent in normal terrestrial gravity exhibited behaviours similar to those of the controls. HG-induced changes in the functional properties of the vestibular system may explain the deficits showed by the HG rats once exposed to normal gravity. The adaptation process to 1 g leading to the appearance of normal behaviour takes about 3 weeks. It likely implicates a central re-evaluation of the sensory inputs and an updating of the motor commands. PMID- 12642181 TI - Relationships between various behavioural abnormalities and nigrostriatal dopamine depletion in the unilateral 6-OHDA-lesioned rat. AB - While rotational asymmetry is used as a characteristic behavioural sign of striatal dopamine (DA) loss in unilateral animal models of Parkinson's disease (PD), there is relatively little analysis of how other common behavioural deficits relate to nigrostriatal DA depletion. We analysed the relationships between several deficits induced by unilateral 6-OHDA lesions and striatal neurochemistry, as well as neuronal loss in the dopaminergic substantia nigra (SN). Behaviour was evaluated from before until 6 weeks after surgery and abnormalities appeared in body axis, head position and sensorimotor performance as well as apomorphine-induced rotation. As expected, rotational behaviour correlated with striatal DA loss and not with other striatal neurotransmitters measured. Similar observations were found for sensorimotor deficits ('disengage task'). Both deficits were observed in rats with >70% loss of TH+ nigral neurons and >80% loss of striatal DA. Additional postural abnormalities appeared with mean losses of 87% of nigral DA neurons and 97% striatal DA, consistent with observations in patients with advanced PD. The data show that the repertoire of behavioural abnormalities manifested by hemiparkinsonian rats relate directly to the degree of nigrostriatal DA loss and, therefore, mimic features of PD. Analysis of such behaviours are relevant for chronic therapeutic studies targeting PD. PMID- 12642182 TI - Visuomotor deficits and fast recovery after area V6A lesion in monkeys. AB - In order to study the involvement of area V6A in visually guided behavior, restricted lesions to the anterior bank of the parieto-occipital sulcus (POs) were made in two adult Cercopithecus aethiops monkeys, trained in visuomotor tests. Beside the known clinical signs of parietal deficit (abnormal posture of the arm contralateral to the most recent lesion and reluctance to use it), misreaching was evident soon after surgery and disappeared quickly. Uncertainties in a landmark test and prolonged response times to the pressing of light-buttons lasted for a few weeks and 1 month, respectively. As suggested by previous anatomical and physiological data, these findings confirm the role of the POs region in the execution of reaching movements. Moreover, they also demonstrate that following lesion, locally networked areas can rapidly adjust in order to re establish pre-lesional behavior. These adjustments take place well before that any anatomical changes may occur. PMID- 12642183 TI - Do 'clumsy' children have visual deficits. AB - Visual processing by 10-year-old children diagnosed on the basis of standardised tests as having developmental 'clumsiness' syndrome, and by a control group of children without motor difficulties, was tested using three different psychophysical tasks. The tasks comprised a measure of global motion processing using a dynamic random dot kinematogram, a measure of static global pattern processing where the position of the target was randomised, and a measure of static global pattern processing in which the target position was fixed. The most striking finding was that the group of clumsy children, who were diagnosed solely on the basis of their motor difficulties, were significantly less sensitive than the control group on all three tasks of visual sensitivity. Clumsy children may have impaired visual sensitivity in both the dorsal and ventral streams in addition to their obvious problems with motor control. These results support the existence of generalised visual anomalies associated with impairments of cerebellar function. PMID- 12642184 TI - Physical and emotional stress have differential effects on preference for saccharine and open field behaviour in rats. AB - Stress may influence the sensitivity of subjects to rewarding stimuli and stress modality may differentially affect this sensitivity. This relation was investigated in our animal model using chronic physical (repeated mild foot shocks) and emotional (witness) stress. Previous research has established that the two stressors have differential long-term effects on behaviour, where physical stress caused inactivity in a small open field and emotional stress hyperactivity. Rats were stressed on 5 consecutive days and tested for locomotor activity in a small open field (day 10) and saccharine preference (day 11). The preference for graded concentrations of saccharine over water was used as a measure for their sensitivity to reward. Physical stress treatment induced a long term decrease both in preference for saccharine and open field activity compared to control treatment. Emotional stress animals showed an increase in open field behaviour activity and a slight increase in saccharine preference. Physical stress seems to cause anhedonia, while emotional stress might cause an increased sensitivity to reward. In conclusion, stress can induce differential long-term effects on sensitivity to positive stimuli and the response to novelty depending on stress modality. PMID- 12642185 TI - Effects of medial prefrontal cortex cytotoxic lesions in mice. AB - Mice (C57BL/6J strain, females) with cytotoxic lesions of the medial wall of the prefrontal cortex were given a battery of tests to assess emotional, species typical, cognitive, motor and other behaviours. Lesioned mice showed a profile of reduced anxiety, both on a plus-maze, and a similar, novel test, the successive alleys. There was no evidence, however, for attenuation of anxiety in tests of hyponeophagia, and lesioned mice, like controls, preferred the black to the white area of an enclosed alley. Their locomotor activity tended to be higher than that of the controls, particularly when the test surroundings were novel or relatively so. Species-typical behaviours were similar to those of control mice, except lesioned mice displaced ('burrowed') less food pellets from a tube in their home cage. They were not impaired at learning a spatial Y-maze reference memory task, which is profoundly affected by cytotoxic hippocampal lesions in the same strain, or at learning a multi-trial passive avoidance test. Their strength and co ordination in motor performance tests was also normal. The results show that cytotoxic medial prefrontal cortex lesions in mice produce a clear but restricted anxiolytic action. The marked reduction in burrowing, in the absence of any detectable impairment of motor ability, demonstrates the sensitivity of this behavioural index. PMID- 12642186 TI - Hemispheric lateralization of memory stages for discriminated avoidance learning in the chick. AB - Memory formation following the one trial discriminated bead task in the chick falls into three stages (short-term, intermediate and long-term memory) that are defined by susceptibility to different classes of drugs. The stages show sharply timed offsets of sensitivity and loss at specific times after inhibition. Recall of the memory in the chick shows cyclical changes that differ in period between left and right hemispheres, and is marked by a series of brief windows of enhanced recall that recur with periods of about 16 and 25 min in the left and right hemispheres respectively. The timing of these 'retrieval events' corresponds, to a large extent, with the timing of the memory stages seen in the visual discrimination task. Here we examine the effects of left or right hemisphere injection of the main agents (glutamate, ouabain and anisomycin) that have been used to characterize the three stages of memory. We show that memory in the left hemisphere is largely responsible for performance at test and that processes involved in its consolidation generate the phases of memory. PMID- 12642187 TI - Behavioural changes after an acute stress: stressor and test types influences. AB - Behavioural consequences of different acute stressors (30 min of restraint, 20 min of forced swim stress, 15 min of inescapable footshocks) applied at the beginning of the active period were assessed in using two behavioural tests: a 20 min light extinction test 24 h after the stressor exposure in order to explore the psychomotor ability and a 10 min open field session within the dark period 48 h after the stressor exposure to estimate the emotional status and the locomotor activity of the rat. Different behavioural responses were observed depending on the nature of the applied stressor. In the light extinction test, the footshock stressed rats developed a very low activity independent on light conditions whereas the rats submitted to forced swim and restraint exhibited an activity level depending on the strain. Moreover, restrained rats had a higher transient activity than forced swim rats under light condition. In the open field test, none of the stressed rats did develop differences in behaviour. The efficacy of a 24 h recovery period on the behavioural response to an acute stressor exposure depends on the intensity of the applied stressor and the behavioural demands. PMID- 12642188 TI - Directional interference during bimanual coordination: is interlimb coupling mediated by afferent or efferent processes. AB - The role of afferent information in bimanual directional interference was studied by means of a modulation of the response-produced information in one of both limbs. In Experiment 1, visual information was either present, withdrawn, or shown with a directional transformation on a LCD screen. In Experiment 2, the technique of muscle tendon vibration was used to bias the kinesthetic afferent information associated with movement. The findings revealed strong evidence for directional interference between both limbs. Nevertheless, no evidence could be advanced that the observed interference from the right onto the left limb movement was modulated by manipulation of the afferent sources of information. It is concluded that directional interference primarily emerges at the efferent level of movement planning and organization. PMID- 12642189 TI - Ventral hippocampal lesions affect anxiety but not spatial learning. AB - Rats with cytotoxic ventral hippocampal lesions which removed approximately 50% of the hippocampus (including dentate gyrus) starting from the temporal pole, displayed a reduction in freezing behaviour following the delivery of an unsignalled footshock in an operant chamber. This was more plausibly a result of reduced susceptibility to fear than a result of a lesion-induced increase in general motor activity. There was no consistent difference between sham and lesioned animals in spontaneous locomotor activity, or locomotion following acute or chronic treatment with amphetamine. In contrast, ventral hippocampal lesioned animals were quicker to pass from the black to the white box during a modified version of the light/dark exploration test, and were quicker to begin eating during tests of hyponeophagia. Furthermore, rats with ventral hippocampal lesions defecated less than their sham counterparts both during open field testing and in extinction sessions following contextual conditioning. In contrast to these clear lesion effects, there were no signs of any spatial learning impairment either in the watermaze or on the elevated T-maze. Taken together these results suggest that the ventral hippocampus may play a role in a brain system (or systems) associated with fear and/or anxiety, and provide further evidence for a distinct specialisation of function along the septotemporal axis of the hippocampus. PMID- 12642190 TI - Massed but not spaced training impairs spatial memory. AB - The Morris water maze and the object displacement task are two popular tools used to investigate spatial learning and memory. Research has focused mainly on the acquisition of spatial tasks while little attention has been given to the retention phase. We examined the effects of different training procedures on retention of the water maze and also reactivity to spatial change in the object displacement task 7 days post-acquisition. We found that massed-trained animals were impaired on retention of the water maze compared to those animals that had received spaced-training. We also found that the massed-trained animals habituated readily to their environment in the object displacement task while the spaced-trained group did not. Furthermore the massed-trained group did not react to spatial change 7 days post-habituation compared to the increased reactivity displayed by the spaced-trained group. Results are discussed in terms of poor encoding of the environment leading to poor retention. PMID- 12642191 TI - Psychosocial and educational aspects in prostate cancer patients. AB - Prostate cancer was a neglected area in psycho-oncology. There is now a growing number of studies on the psychosocial aspects of having prostate cancer and the possibilities to reduce these problems in educational and group interventions. In this issue of Patient Education and Counseling, studies are presented on several psychosocial and educational aspects in prostate cancer patients: screening events and outcomes, assessing the unmet information, support and care delivery needs, reacting to the diagnosis of prostate cancer, informational needs of men on hormonal therapy, changes in health-related quality of life three months after the diagnosis, information-seeking behaviors and information needs of partners, quality of leaflets, video information in decision making, and patient perceptions and priorities in a rehabilitation program. Conclusions are presented on neglected research areas in psychosocial and educational aspects of living with prostate cancer. PMID- 12642192 TI - Interpretation of prostate cancer screening events and outcomes: a focus group study. AB - Professional guidelines call for physicians to provide patients with information to permit informed decision making. We conducted focus groups to obtain reactions to numerical information about events and outcomes related to prostate cancer screening (prevalence, natural history, accuracy of screening, and treatment outcomes). The focus groups were used to help develop a balance sheet, a decision aid that explicitly compares likelihood and value of outcomes. In all, 90 persons participated in 12 focus groups homogeneous in ethnicity (African American, White, Hispanic) and gender. Discussions were transcribed and analyzed using qualitative methods. The view of screening derived by participants from the numerical information was less positive than initial opinions based on the media and confidence in medicine. Participants suggested shorter screening intervals, regular screening, and different treatment methods. No single topic was seen as sufficient for the screening decision. The balance sheet should cover prevalence and outcomes of screening and treatment. PMID- 12642193 TI - Reacting to the diagnosis of prostate cancer: patient learning in a community of practice. AB - The diagnosis of prostate cancer brings with it a number of emotional issues for the patient. These may include: fear of cancer; confronting one's own mortality; quality of life issues such as incontinence and impotence; and the question "why me?" In addition, there is burden of coping with the emotional responses from friends and loved ones which, arising from their concern, can range from fear to seeking to take charge. Added to this is the need to make choices regarding treatment and obtaining information about these. This article is based on a contribution to a symposium on prostate cancer at the Fifth International Conference on Psycho-Oncology and describes a personal experience of confronting these issues. The concept of communities of practice is used to interpret the learning experiences undertaken by the cancer patient. PMID- 12642194 TI - Changes in health-related quality of life of men with prostate cancer 3 months after diagnosis: the role of psychosocial factors and comparisment with benign prostate hyperplasia patients. AB - The objective of the pilot study is to measure the changes in health-related quality of life (HRQOL, general and prostate specific) 3 months after the disease has been diagnosed in men with prostate cancer. The results are compared with benign prostate hyperplasia (BPH) patients. Also the influence of psychosocial factors (distress, coping and social support) on the experienced changes in health-related quality of life is studied for both patient groups. The sample consists of 61 patients who filled in the questionnaire before the diagnosis was know to them and after 3 months (38 benign prostate hyperplasia and 23 prostate cancer patients). The measures consist of: quality of life, micturation symptoms, sexual functioning, coping style, psychological distress, life events, social support, social desirability, and health behavior. The results show a decrease in quality of life for prostate cancer patient after three moments, while the quality of life measure for BPH patients are stable over time. For both patient groups, the psychosocial factors do not change after 3 months. The psychosocial factors contribute only marginal to the changes in quality of life. Conclusions are drawn concerning the need and the content of psychosocial support and education for prostate cancer patients. PMID- 12642195 TI - Assessing the unmet information, support and care delivery needs of men with prostate cancer. AB - This study identified the key Unmet Needs of men with localized prostate cancer. A series of Nominal Groups were used to identify needs, from which a 135-item survey was developed to assess both the Importance and Unmet Need of each item. An Importance-Weighted Unmet Need score was calculated for each item, incorporating both the Importance and the degree to which the need was unmet. Surveys (n=500) were distributed in four geographically distinct areas, with a response rate of 46%. Respondents were 90% Caucasian, 80% married, with a mean age of 66 years, and mean education of 14 years. Care delivery needs were most important and least unmet, while Support needs were least important and most unmet. However, when degree to which needs were unmet was weighted by Importance, information needs had the highest Importance-Weighted Unmet Need scores. The greatest Unmet Needs for information were in knowledge of recurrence issues and in side effects of the illness and its treatment. PMID- 12642196 TI - Informational needs of men with prostate cancer on hormonal manipulation therapy. AB - Prostate cancer is a significant health care issue worldwide and the global incidence of this disease is predicted to treble by 2018. The mainstay of treatment for men with locally advanced prostate cancer is hormonal manipulation therapy (hmt), which has undesirable side effects such as impotence. To help individuals cope with a diagnosis of cancer, research has found that specific information is required. A descriptive exploratory study was undertaken to assess the knowledge and informational needs of a sample of men with locally advanced prostate cancer on hmt in the United Kingdom (UK). Results illustrated that these men had knowledge deficits coupled with a great desire for information regarding their disease and treatment. This study identified that these men required more information and it helps to address the apparent neglect of this subject in the domain of prostate cancer research. Through recognition of the informational needs of this patient group, methods of delivering evidence-based educational interventions in the clinical domain could be promoted. PMID- 12642197 TI - The relationship between the information-seeking behaviours and information needs of partners of men with prostate cancer: a pilot study. AB - Although information can have many benefits for individuals with cancer, information-seeking theories suggest that some individuals fail to benefit from voluminous information. Therefore, it is important that the information-seeking behaviours of patients and their families are taken into consideration when identifying their needs for information. This pilot study investigated the relationship between the information-seeking behaviours and information needs of partners of men with prostate cancer. Thirty-nine partners of men with prostate cancer completed the study instruments, which identified their information seeking behaviours and their information needs. Significant and positive correlations were found between participants' information-seeking behaviours and their information needs, suggesting that as individuals' propensities to seek information increased so too did their needs for information. The results suggest that researchers and healthcare professionals need to take into consideration the information-seeking behaviours of patients and their family members when identifying their information needs. PMID- 12642198 TI - Patient information leaflets for prostate cancer: which leaflets should healthcare professionals recommend? AB - This study evaluated 31 patient information leaflets (PILs) discussing treatment options for prostate cancer. In stage one, the authors evaluated the leaflets' quality, readability and suitability using objective measures: the DISCERN instrument; Flesch formula; and the Suitability Assessment of Materials (SAM) instrument, respectively. Although the leaflets varied in terms of their scores on each measure, it was possible to identify the best five leaflets across the three conditions. In stage two, eight men with prostate cancer took part in a focus group discussion or individual interview to outline their views regarding a number of leaflets, including the best five booklets or leaflets identified in stage one of the study. The interviews were audiotaped and analysed using a template analysis. Patients were able to discriminate between the best five leaflets or booklets and identify their preferred booklets. These were the booklets: "Understanding cancer of the prostate" by CancerBACUP, "Prostate cancer: everything you need to know" by the Prostate Cancer Charity and "The treatment of prostate cancer. Questions and answers" by the Covent Garden Cancer Research Trust. On the basis of their high ratings for the objective measures and patients' views, healthcare professionals are advised to recommend these three booklets to men with prostate cancer who want written information about the disease. However, randomised controlled trials are needed to examine the impact of these booklets on the psychosocial outcomes of men with prostate cancer. PMID- 12642199 TI - What information patients with localised prostate cancer hear and understand. AB - Patients with localised prostate cancer can decide on a treatment option that is appropriate for their needs and lifestyle only if their doctor provides information that they understand and can remember. This small study (n10) examined the knowledge of men who had been diagnosed but had not been treated for early stage disease. A semi-structured interview schedule enabled these men to demonstrate their knowledge of their disease and its treatment. They were aware that they had incomplete understanding, but no one asked that information should be provided until, and in a language, they understood. Only two men felt that the choice of treatment belonged to them, others agreed with the decision of their urologist although they did not understand why a specific treatment was recommended or the implications of that treatment. PMID- 12642200 TI - Information on video format can help patients with localised prostate cancer to be partners in decision making. AB - A video was produced to inform men diagnosed with localised prostate cancer of the different management options for their disease. It was tested for increased knowledge and understanding on a healthy population of similar age (n=10), who were assessed before and after watching the video (20 interviews). The video was then shown to 12 men who had been diagnosed with prostate cancer but had not started any treatment for their disease (24 interviews). Interviews among the healthy cohort revealed not only the lack of knowledge of anatomy and physiology but also the way in which these men used past experiences to help explain and remember complex medical procedures. The patients focused on the treatments and outcomes, remembering less of the technicalities. All men interviewed vividly remembered the patients participating in the video; they drew inferences and developed affinity towards specific individuals. Both groups felt that video provided information in a user friendly way. Although remembering few anatomical terms all participants described a visual image that led to a perception of understanding rather than a definitive increase in knowledge. Patients, however, felt they had a clearer understanding of the disease and its treatment, and therefore better able to be active informed partners in the decision-making process. PMID- 12642201 TI - "Between men": patient perceptions and priorities in a rehabilitation program for men with prostate cancer. AB - The objective of the study is to compare consumer aspects of an informative, a physical, and a combined informative and physical rehabilitation program included in the "Between men" project for newly diagnosed prostate cancer patients. A consecutive series of patients was randomized. Programs were especially developed for prostate cancer patients. The format was 7 weekly sessions. The results show that the perceived benefits of relaxation was greater in the combination group than in the physical training group only. In comparison with the physical group more patients in the informative groups (information and information+physical training) rated the knowledge received as very important. The majority of patients (90%) was of the opinion that the "Between men" programs, should be continued. Independent of the actual program given, patients opted for the combination program or information alone but not the physical training alone program. PMID- 12642227 TI - The role of nitric oxide in oral diseases. AB - Although previously regarded as a toxic pollutant gas, nitric oxide (NO) is a short-lived molecule that plays a key role in many physiological and pathological processes. It is produced in vivo from the amino acid L-arginine by a complex family of enzymes termed nitric oxide synthase (NOS). Since its discovery as a biological messenger in 1987, NO has been implicated in many disease processes, ranging from septic shock to cancer. It is a highly reactive free radical and causes concentration-dependent conformational changes in proteins, enzymes and DNA, predominantly by its reaction with transition metals and thiol residues. Although high concentrations of NO are cytotoxic, the levels produced in many human cancers possibly facilitate tumour growth and dissemination. The interest in this molecule by scientists and clinicians involved with the oral cavity and head and neck regions is fairly recent, and only a tiny minority of 50,000 papers currently cited on NO relate to diseases in this anatomical area. This review gives an overview of NO, outlining its basic chemistry, formation by NOS and its possible roles in the oral diseases studied to date. The implications for possible therapeutic manipulation of NO are also discussed. PMID- 12642228 TI - Glycogen content and activities of enzymes involved in the carbohydrate metabolism of the salivary glands of rats during postnatal development. AB - Carbohydrate metabolism was examined in the developing rat salivary glands by analysing enzymatic activity and glycogen content in the postnatal parotid and submandibular glands. The following enzymes of the carbohydrate metabolism, hexokinase (HK), phosphofructokinase-1 (PFK-1), pyruvate kinase (PK), glucose-6 phosphate dehydrogenase (G6PD), and lactate dehydrogenase (LDH) as well as the content of glycogen were determined in the salivary glands of rats aged 2, 7, 14, 21, 30 and 60 days. The specific activity of HK increased from days 2 to 21 and then it decreased up to 60 days old. The values found for the submandibular glands were from 2.5 to 4.9 times higher than those found for the parotid gland, except for rats aged 60 days. PFK-1 showed a different pattern of variation between the glands. In the submandibular gland there was a statistically significant increase in PFK-1 specific activity from 2 to 30 days of age and then, in the 60 days old group a return to level of the rats aged 2 days. In parotid gland, the specific activity of PFK-1 decreased between 2 and 7 days of age, from 7 to 14 days the specific activity increased markedly and from 14 to 60 days old it gradually decreased. The specific activity of PK followed the same pattern of variation in the submandibular and parotid glands, showing no great variation. The specific activity of LDH decreased from 2 to 60 days old in the submandibular glands. In the parotid glands the mean values for this enzyme were higher for the 2 days old group, and then decreased to remained more or less constant. The potential capacity of the pentose phosphate pathway was greater than that of glycolysis at early ages. The glycogen content showed similar variation in both glands. It was initially high and then decreased. In conclusion, our results on the activities of enzymes involved in carbohydrate metabolism in submandibular and parotid glands may be relevant to the initiation of saliva secretion in these animals. PMID- 12642229 TI - Transforming growth factor beta in chronic obstructive sialadenitis of human submandibular gland. AB - BACKGROUND: The exact pathomechanism of inflammation progress and fibrosis in chronic obstructive sialadenitis is unknown. The aim of the present study was to assess whether there is an association between transforming growth factor beta (TGF-beta) and fibrogenic process of chronic sialadenitis. METHODS: Tissue samples of 12 patients with chronic sialadenitis and 4 normal tissue samples of the submandibular gland were examined immunohistochemically for identification of TGF-beta. TGF-beta1 messenger RNA (mRNA) expression was analysed semiquantitatively using reverse transcription polymerase chain reaction and gel electrophoresis to correlate its expression levels with stages of the disease. RESULTS: TGF-beta positive cells could be found in the secretory duct system of all examined samples. However, an intense TGF-beta immunoreactivity was observed in inflamed salivary glands. With progress of disease TGF-beta1 mRNA expression increases significantly. CONCLUSION: Expression of TGF-beta in chronic sialadenitis and its apparent increase in advanced stages of the disease, suggests that this growth factor may play a role in glandular fibrosis. PMID- 12642231 TI - Cell-cycle regulatory proteins in human wound healing. AB - Proper healing of mucosal wounds requires careful orchestration of epithelial cell migration and proliferation. To elucidate the molecular basis of the lack of cellular proliferation in the migrating 'epithelial tongue' during the re epithelialization of oral mucosal wounds, the expression of cell-cycle regulators critical for G(1)-phase progression and S-phase entry was here analysed immunohistochemically. Compared to normal human mucosa, epithelia migrating to cover 2- or 3-day-old wounds made either in vivo or in an organotypic cell culture all showed loss of the proliferation marker Ki67 and cyclins D(1) and A, and reduced expression of cyclins D(3) and E, the cyclin D-dependent kinase 4 (CDK4), the MCM7 component of DNA replication origin complexes and the retinoblastoma protein pRb. Among the CDK inhibitors (CKIs), p16ink4a and p21Cip1 were moderately increased and decreased, respectively, whereas the abundance of most of the CKIs, including p27Kip1, p57Kip2, p15ink4b and p18ink4c, was relatively maintained in the migrating epithelial tongue. These data indicate that downmodulation of several G(1)/S-phase cyclins and a relative excess of CKIs may cooperate to ensure the quiescent state of migrating keratinocytes during wound healing. PMID- 12642230 TI - Oral colonisation by aerobic and facultatively anaerobic Gram-negative rods and yeast in Tibetans living in Lhasa. AB - Sample groups of children (n=50) and adults (n=38) were selected from pools of 207 children, (11-13-year olds from two primary schools) and 94 adults (25-44 year olds from four governmental agencies) who were the subjects of an oral health survey among Tibetans living in Lhasa, Tibet Autonomous Region. Mean ages of the study groups of children (38% females) and adults (61% females) were 11.6+/-0.9 and 37.1+/-6.1 years, respectively. All had lived in Tibet since birth. Oral rinse samples were selective cultured to isolate, quantify and speciate aerobic and facultatively anaerobic Gram-negative rods (using the API 20E kit) and yeasts (using API 20C AUX and API ZYM kits). For children, the isolation rates for oral coliform bacteria and yeasts were 84 and 14%, respectively, for adults, the respective rates were 26 and 40%. The corresponding quantities of coliforms/yeasts for children and adults were 0.4+/-1.6 x 10(3)c.f.u./15.8+/-72.3 and 0.2+/-0.6 x 10(3)c.f.u./57.2+/-137.5c.f.u. per millilitre oral rinse, respectively. Aerobic and facultatively anaerobic Gram negative rods and Stenotrophomonas maltophilia, a free-living saprophytic and ubiquitous bacterial species of wide geographic distribution, were significantly more frequently recovered from the children's oral rinses. The isolation rates of facultatively anaerobic Gram-negative rods in adults and yeasts in both groups were similar to those found in similar cohorts from southern China in earlier studies. Randomly amplified polymeric DNA analysis showed that the S. maltophilia spp. isolated from children were of several different clonal types and were school specific. This study shows that the colonisation rate of facultatively anaerobic Gram-negative rods in adults and yeasts in both groups are similar to those in populations living at lower altitudes, the native young, urban Tibetans appear to exhibit a high oral carriage rate of S. maltophilia spp. PMID- 12642232 TI - Modulation of SOD activity in rat submandibular glands. AB - Reactive oxygen species (ROS) such as superoxide anion (O(2)(*-)), hydrogen peroxide (H(2)O(2)) and hydroxyl radical (*OH) are produced constantly in aerobic organisms. Submandibular glands of rats treated with isoproterenol (ISO) were analysed in respect to the level of superoxide dismutase (SOD), lipoperoxidation, protein oxidation and glutathione (GSH)/GSSG. Although ISO response is very well studied in salivary glands cells, the effect of ISO treatment on ROS productions or on SOD activation is not known. SOD activity increased after isoproterenol subcutaneous injection. In contrast, cycloheximide, a protein synthesis inhibitor, was found to decrease the total SOD activity. The GSH/GSSG ratio and the lipoperoxidation were also found to be increased after isoproterenol injection, while the protein oxidation was not affected by the same treatment. Injection of isoproterenol after cycloheximide treatment resulted in the recovery of the total SOD activity. This increase of activity is related to the amount of MnSOD isoform as shown by Western blot analysis. Our results suggest that the antioxidant protection of SOD may be activated during the salivary glands ISO treatment and that the mitochondrial MnSOD has an important role in the oxidative stress process. PMID- 12642233 TI - In vitro action of Bordeaux red wine on the microhardness of human dental enamel. AB - Several studies have demonstrated that the regular and large consumption of wine is associated with increased risk of tooth erosion. Here, the effect of Bordeaux red wine on enamel was estimated by measuring changes in its Vickers microhardness. Thirty premolars were used; microhardness tests were performed on buccal areas before and after 10, 30, 90 and 120 s immersion in the wine (pH=3.9). Enamel surfaces were also observed by scanning electron microscopy. No statistically significant difference was found between the mean Vickers microhardness obtained at t=0 and 90 s, but slight signs of enamel demineralisation were observed with the scanning electron microscope. It appears that wine has no disastrous effect on the microhardness of dental enamel when the two are in contact for less than 90 s. When the exposure is for at least 120 s, it may become harmful, as the decrease in the microhardness of enamel was then significant (P<0.05). PMID- 12642235 TI - Microradiography to evaluate bone growth into a rat mandibular defect. AB - Microradiography has been evaluated to measure bone healing into a 5.0mm outer diameter mandibular defect in the rat. This method provides high-resolution radiographs of the defects that can be used for an accurate measurement of bone defect healing. In 12 rats, the defect widths of 42-day-old mandibular defects have been measured both using microradiographs and histological sections. The defect width+/-S.D. measured 3.42+/-0.98 mm microradiographically and 3.47+/-1.11 mm histologically. Both methods were accurate in determining defect widths but microradiography has the advantage over histology that an image is obtained from the entire defect, making it possible to measure areas of bone growth. PMID- 12642234 TI - Patterns of secretion of mucins and non-mucin glycoproteins in human submandibular/sublingual secretion. AB - The present investigation has characterised the influence of gustatory stimulation and duration of stimulation on the secretion pattern of salivary mucins MG1 and MG2 and non-mucin glycoproteins in submandibular/sublingual secretion (SMSL). Resting SMSL was collected for three 2 min intervals and stimulated SMSL was collected for ten 1 min intervals from six healthy subjects. Flow rates and total protein were significantly different under the two conditions. The secretion patterns of these proteins under resting and stimulated conditions was examined on periodic acid-Schiff reagent (PAS)-stained polyacrylamide gels using a Kodak Digital-Science Image Station. Image analyses revealed that the level of MG1 increased and the level of MG2 remained nearly the same after stimulation. Six other major glycoproteins (designated Band 1-6) were identified on the basis of their electrophoretic mobilities and immuno-reactivity on Western blots. After stimulation the intensity of Band 1 (lactoferrin and peroxidase) and Band 2 (amylase) decreased whereas the intensity of Band 3 (carbonic anhydrase), Band 4 (proline-rich glycoprotein) and Bands 5 and 6 (basic glycosylated proline-rich proteins) increased. These patterns probably reflect secretion from preformed vesicles since de novo synthesis would be unexpected within the time frame of these experiments. The variable patterns observed suggest that mucins and non-mucin glycoproteins in SMSL derive from different subsets of secretory vesicles, some of which may originate in mucous and others in serous acini, as well as in ductal cells. Quantification of mucins was performed by image analysis technology using purified MG1 and MG2 standards. Finally, the present investigation has shown that the secretory patterns of mucins and non-mucin glycoproteins from submandibular/sublingual glands are complex and represent an important aspect of salivary gland physiology. PMID- 12642236 TI - Inflammatory hypersensitivity in a rat model of trigeminal neuropathic pain. AB - Thermal and mechanical stimuli have been used to monitor the development of neuropathic pain following an experimental injury to a branch of the trigeminal nerve. However, the response to inflammatory challenge has not been evaluated in a model of orofacial neuropathic pain. The purpose of this study was to determine whether chronic constriction of the infraorbital nerve (IoN) enhances nociceptive responses elicited by the formalin test. The characteristic biphasic response (primarily directed grooming) to formalin injected subcutaneously in the right vibrissae pad was observed in sham-injury rats. Twenty-one days after IoN constriction, formalin injection provoked an immediate response that involved both directed grooming and other abnormal behaviors, e.g. flinching, trismus and shielding of the affected region. As with sham-injury rats, this was followed by a quiescent period and then a second phase of nocifensive behaviors. The total time recorded for all pain-related behaviors was significantly greater in rats with constrictive injuries (P<0.001), due primarily to the exhibition of novel pain-related responses. Histological examination (qualitative) revealed that chronic constriction resulted in a ligature-induced neuroma, as well as a partial denervation of the affected sensory field. Thus, an intense inflammatory hypersensitivity in a rat model of orofacial neuropathic pain develops in association with partial denervation and with an ongoing perineural inflammatory response and neuroma formation. PMID- 12642237 TI - Expression of MIP-3alpha/CCL20, a macrophage inflammatory protein in oral squamous cell carcinoma. AB - We have examined the expression of MIP-3alpha/CCL20 in oral squamous cell carcinoma (SCC) in vivo and in vitro. In addition, we have investigated whether the expression of MIP-3alpha/CCL20 is regulated by bacterial infection and inflammatory cytokines. In order to determine the mRNA level of MIP-3alpha, quantitative reverse transcription-polymerase chain reaction (RT-PCR) was performed with LightCycler using the double-stranded DNA dye, SYBR Green I. Oral epithelial cells and six SCC cell lines (SCC-9, SAS, BSC-OF, HSC-4, HSC, Ca9-22) were found to express MIP-3alpha mRNA. The expression of MIP-3alpha was upregulated by infection with Actinobacillus actinomycetemcomitans and by stimulation with lipopolysaccharide and TNF-alpha. By in situ hybridization, the detectable MIP-3alpha expression in SCC was localized primarily at the epithelial pearls corresponding to the spinous layer. These results suggest that MIP-3alpha contributes to the oral immunoresponse to bacterial infection, and may be involved in the growth of SCC. PMID- 12642238 TI - Methyl jasmonate induces changes mimicking anatomical defenses in diverse members of the Pinaceae. AB - Conifers have defenses such as the production of phenolic compounds and resins that can be induced by bark beetles and other invading organisms, but the signaling agents involved are unknown. The anatomical effects of methyl jasmonate (MJ), a potent inducer of certain plant defenses, were compared with wounding of the bark of 12-15-year-old trees of five conifer species. Wounding in all species resulted in tissue necrosis and wound periderm development immediately around the wound site. One cm from the wound, swelling of phloem polyphenolic parenchyma cells and phenolic accumulation were observed in Pseudotsuga menziesii (Mirb.) Franco, Picea pungens Engelman, Larix occidentalis Nutt. and Pinus monticola Douglas ex D. Don, but not in Taxus brevifolia Nutt. Traumatic resin ducts were formed in response to wounding in three species of Pinaceae, but not in P. monticola, which formed irregular clusters of cells rather than ducts. Taxus brevifolia did not form resin ducts in response to either wounding or MJ treatment. In the Pinaceae species studied, surface application of 100 mM MJ caused similar anatomical changes to those observed in response to wounding, including phenolic accumulation, cell swelling and traumatic resin duct formation, but it did not induce a wound periderm. Traumatic resin ducts differed in size among the study species, ranging from small in L. occidentalis to very large in P. menziesii. In P. menziesii, P. pungens and L. occidentalis, traumatic resin ducts were more abundant after MJ treatment than after wounding. We conclude that the octadecanoid pathway is likely involved in defense responses in stems of the Pinaceae, but not necessarily in other taxa. PMID- 12642239 TI - Changes in fruit sugar concentrations in response to assimilate supply, metabolism and dilution: a modeling approach applied to peach fruit (Prunus persica). AB - The influence of assimilate supply, metabolism and dilution on sugar concentrations in the mesocarp of peach (Prunus persica (L.) Batsch) fruit during the main stage of fruit enlargement was analyzed with the SUGAR model of Genard and Souty (1996). The model predicts the partitioning of carbon into sucrose, sorbitol, glucose and fructose in the mesocarp of peach fruit. Based on measured data and the model, we determined values for the relative rates of sugar transformation. These rates were constant, varied with time or varied with relative fruit growth rate, depending on the type of sugar. Equations were derived to describe these rates and incorporated into the SUGAR model. The model simulated the effects of changing assimilate supply and fruit volume on sugar concentrations. The set of equations for the SUGAR model was rewritten to include the three components influencing sugar concentrations: assimilate supply, metabolism and dilution. The sugar types differed in sensitivity to these components. Sucrose was highly sensitive to changes in assimilate supply and to the dilution effect; it was not subject to intense metabolic transformation. Sorbitol was the most important carbohydrate in fruit metabolism, which explains why the sorbitol concentration was always low despite the strong positive effect of assimilate supply. The reducing sugars constituted a transitory storage pool and their concentrations were closely related to metabolism. PMID- 12642240 TI - Effects of tension wood on specific conductivity and vulnerability to embolism of Quercus ilex seedlings grown at two atmospheric CO2 concentrations. AB - To determine whether there are decreases in hydraulic function of a woody stem when it has increased mechanical loading, Quercus ilex L. seedlings were grown upright or inclined to force the production of large amounts of tension wood (TW). Seedlings were grown in ambient or elevated carbon dioxide concentrations ([CO2]) for 16-17 months to provide two sets of seedlings differing in growth rates and allocation patterns. In both CO2 environments, inclination caused formation of large amounts of TW at the base and mid-section of most stems, but not at the stem tips. Contrary to expectation, there were no significant effects of stem inclination or amount of TW on specific conductivity (k(s)) or vulnerability to embolism. Samples with high amounts of TW had higher vessel frequency, similar average vessel lumen area, similar vessel lumen fraction (6% of the transverse area), elevated frequency of vessels in the smallest diameter class, and higher wood density than samples with very little TW. Samples from seedlings in the elevated [CO2] treatment had similar vessel frequency, larger average vessel lumen area (caused by a higher frequency of large-diameter vessels), similar vessel lumen fraction, and similar wood density as samples from seedlings in the ambient [CO2] treatment. There was a strong position effect: the highest wood density and lowest ks were at the stem base, intermediate values were at the middle, and the lowest density and highest ks were at the stem tip. We conclude that, in a species that uses different cells for mechanical support and water transport, there can be large modifications in performance of the mechanical function through TW formation without impacting the water transport functions-ks and vulnerability to embolism. PMID- 12642241 TI - Osmotic adjustment in three-year-old seedlings of five provenances of maritime pine (Pinus pinaster) in response to drought. AB - Three-year-old seedlings of five provenances of Pinus pinaster Ait. that differed in climatic conditions at their geographical origin were subjected to decreasing soil water availability. The degree of needle osmotic adjustment (OA) was estimated based on logarithmic plots of needle relative water content (RWC) against needle osmotic potential (Psi(pi)); i.e., lnRWC versus -ln(-Psi(pi)). There were significant differences among provenances in active OA (0.13 to 0.30 MPa for a decrease in RWC to 80%), and a clear negative relationship was found between OA and precipitation (650 to 1280 mm of mean annual rainfall) at the geographical origins of the provenances. A high osmoregulatory capacity contributes to the maintainance of positive turgor at low water potentials. We conclude that OA is one of the mechanisms underlying adaptation to drought in P. pinaster. Solute accumulation was about 2.3 times higher in the provenance from the driest site than in the provenance from the wettest site. The contribution of osmotic adjustment to differences in drought tolerance mechanisms among provenances is discussed. PMID- 12642242 TI - Bacterial disease resistance of transgenic hybrid poplar expressing the synthetic antimicrobial peptide D4E1. AB - Important losses in poplar productivity occur because of susceptibility to microbial pathogens. To enhance disease resistance in susceptible genotypes, the gene coding for D4E1, a synthetic antimicrobial peptide consisting of 17 amino acid residues, was introduced into poplar (Populus tremula L. x Populus alba L.) via Agrobacterium-mediated transformation. Four kanamycin-resistant transformants were selected based on significant accumulation of the D4E1 transcript and confirmed by reverse transcription-polymerase chain reaction and RNA dot-blot analysis. These transgenic poplar lines were tested for resistance to Agrobacterium tumefaciens, Xanthomonas populi pv. populi and Hypoxylon mammatum (Wahl.) Miller. One transgenic poplar line, Tr23, bearing the highest transcript accumulation for the D4E1 gene, showed a significant reduction in symptoms caused by A. tumefaciens and X. populi. However, none of the transgenic poplar lines showed a significant difference in disease response to the fungal pathogen H. mammatum. PMID- 12642243 TI - Timing of GA4/7 application and the flowering of Pinus sylvestris grafts in the greenhouse. AB - Effects of varying the time of stem injection of gibberellin A4/7 (GA4/7) on male and female flowering in potted grafts of Pinus sylvestris L. in a greenhouse were monitored in two consecutive years. Flowering results for each application time were correlated with phenological characteristics and abiotic factors in an attempt to find reproducible indicators of the most effective time of GA4/7 treatment. The GA4/7 treatment had a significant effect on female flowering, but no effect on male flowering. In both years, the largest effect on female flowering was obtained with the latest application. Percent shoot and needle elongation (relative to final length) were poor indicators of the optimal time of GA4/7 application. Three heat sum accumulation models gave equally accurate predictions of the optimal time for GA4/7 application; therefore, the simplest model, i.e., the sum of degree-days, is recommended. The timing window for stimulation of female flowering with GA4/7 had a lower limit of about 500 degree days (threshold +5 degrees C), and the optimal time of GA4/7 application was after about 700 degree-days. PMID- 12642244 TI - Somaclonal variation in Coffea arabica: effects of genotype and embryogenic cell suspension age on frequency and phenotype of variants. AB - We determined how age of embryogenic cell suspensions affects somaclonal variation in five F1 hybrids of Coffea arabica L. Batches of plants were produced either directly from embryogenic callus, or after 3, 6, 9 and 12 months of embryogenic cell suspension culture. Seven phenotypic variants were characterized. Based on vigor and productivity of the regenerated plants, we classified the variants in order of increasing severity of physiological disorders as: Juvenile leaf color, Giant, Dwarf, Thick leaf (Bullata), Variegata, Angustifolia, and Multi-stem. The Dwarf, Angustifolia and Multi-stem variants were the most frequent among the regenerated plants (1.4, 4.8 and 2.9%, respectively). The frequency (f) of variants increased exponentially with the age (t) of the embryogenic suspension, in accordance with the function f = 0.99e(0.267t). For all genotypes, somaclonal variation was low (1.3%) in plants produced from embryogenic callus or 3-month-old cell suspensions and increased in frequency with increasing suspension age (6, 10 and 25% in plants produced from cell suspensions aged 6, 9 and 12 months, respectively). Large differences in somaclonal variation among genotypes were found only in plants produced from 12 month-old cell suspensions. For two genotypes, the oldest suspensions produced a majority of somaclonal variants (80-90%), whereas somaclonal variation ranged between 8 and 18% in the other genotypes. Cell suspension age and genotype also affected the type of variant produced. The severity of somaclonal variations increased with cell suspension age. For all genotypes combined, the Angustifolia variant was the most common. The other somaclonal variations were specific to certain genotypes or distributed randomly among the genotypes. PMID- 12642245 TI - Growth response of cottonwood roots to varied NH4:NO3 ratios in enriched patches. AB - Maximization of short-rotation forest plantation yield requires frequent applications of nutrients, especially nitrogen (N). Whole-plant growth is known to be sensitive to the proportion of ammonium to nitrate (NH4:NO3). However, the extent to which N form affects root growth, branching and morphology is poorly understood, and these variables may have substantial impacts on plant nutrient and water acquisition. We used rooted cuttings of cottonwood (Populus deltoides Bartr. ex Marsh.) to investigate the effect of various NH4:NO3 ratios on root growth in N-enriched patches. A sand culture study with split-root systems was carried out in which 1-3% of the total root system of each cutting was supplied with 2 mM N at NH4:NO3 ratios of 0:100, 20:80, 40:60, 60:40, 80:20 and 100:0 (molar basis), with the rest of the plant supplied with 0 mM N, resulting in the whole plant becoming N deficient. During the experiment, whole-plant growth was unaffected by the treatments. Of the NH4:NO3 ratios tested, greatest total root length, specific root length, and root N concentration of roots in enriched patches occurred in the 20:80 NH4:NO3 treatment. The largest response of roots in enriched patches was third- and fourth-order root production. We conclude that N form has a profound effect on root development and morphology in enriched patches. PMID- 12642247 TI - Anterior cruciate ligament rupture after thermal treatment in a canine model. AB - BACKGROUND: The use of radiofrequency energy to treat damaged anterior cruciate ligaments is gaining popularity. However, complete rupture of the ligament after treatment has been reported. PURPOSE: To evaluate the effect of thermal energy applied arthroscopically to normal, intact anterior cruciate ligaments in mature dogs. STUDY DESIGN: Controlled laboratory study. METHODS: Monopolar radiofrequency energy was applied to the normal anterior cruciate ligament of 1 knee in 18 dogs. The contralateral anterior cruciate ligament (also normal) was sham treated. Force-plate gait analysis was performed preoperatively and at 4, 8, 12, 16, 26, and 36 weeks after surgery. Anterior cruciate ligament rupture was detected by a sudden onset of nonweightbearing and a positive drawer sign. RESULTS: All treated ligaments ruptured approximately 55 days after surgery (mean, 55 days; standard error, 1.6). CONCLUSIONS: Although monopolar radiofrequency energy may have some potential in the treatment of lax anterior cruciate ligaments, in the application described here the result was a highly predictable deterioration and rupture of all treated anterior cruciate ligaments. CLINICAL RELEVANCE: On the basis of these findings, we strongly recommend that strict selection and application criteria be used when considering use of this modality on anterior cruciate ligaments that are stretched or partially disrupted, or both. Use of this modality should be followed by adherence to a highly conservative rehabilitation protocol. PMID- 12642248 TI - Relationship between postconcussion headache and neuropsychological test performance in high school athletes. AB - BACKGROUND: The relevance of headache to outcome after sports-related concussion is poorly understood. HYPOTHESES: High school athletes reporting headache approximately 1 week after injury will have significantly more other concussion symptoms and will perform more poorly on neuropsychological tests than athletes not experiencing headache. STUDY DESIGN: Prospective cohort study. METHODS: Study participants included 109 high school athletes who had sustained concussion and who were divided into two groups: those reporting headache 7 days after injury and those reporting no headaches. The two groups were compared regarding on-field markers of concussion severity at the time of injury and symptoms and neurocognitive test results collected via ImPACT, a computerized neuropsychological test battery and postconcussion symptom scale, at a mean of 6.8 days after injury. RESULTS: Athletes reporting posttraumatic headache demonstrated significantly worse performance on reaction time and memory ImPACT neurocognitive composite scores. These athletes also reported significantly more symptoms other than headache and were more likely to have demonstrated on-field anterograde amnesia. CONCLUSIONS: Findings suggest that any degree of postconcussion headache in high school athletes 7 days after injury is likely associated with an incomplete recovery after concussion. PMID- 12642249 TI - The fixation strength of six hamstring tendon graft fixation devices in anterior cruciate ligament reconstruction. Part I: femoral site. AB - BACKGROUND: Strength of graft fixation is the weakest link in anterior cruciate ligament reconstruction. HYPOTHESIS: There is no difference in initial fixation strength between different hamstring tendon graft femoral fixation devices. STUDY DESIGN: Randomized experimental study. METHODS: Each of six devices was used in the fixation of 10 quadrupled human semitendinosus-gracilis tendon grafts in tunnels drilled in porcine femora and tested 10 times with a single-cycle load-to failure test at a rate of 50 mm/min and 10 times with a 1500-cycle loading test between 50 and 200 N at one cycle every 2 seconds. The specimens that survived the cyclic loading were subjected to a single-cycle load-to-failure test. RESULTS: The Bone Mulch Screw (1112 N) was strongest in the single-cycle load-to failure test, followed by EndoButton CL (1086 N), RigidFix (868 N), SmartScrew ACL (794 N), BioScrew (589 N), and RCI screw (546 N). It also showed the lowest residual displacement (2.2 mm) and was strongest in the single-cycle load-to failure test after cyclic loading. CONCLUSIONS: The Bone Mulch Screw was superior to all other devices. CLINICAL RELEVANCE: Caution may be warranted in employing aggressive rehabilitation after reconstruction with these devices. Preconditioning of the graft-implant complex before fixation is important. PMID- 12642250 TI - The fixation strength of six hamstring tendon graft fixation devices in anterior cruciate ligament reconstruction. Part II: tibial site. AB - BACKGROUND: Tibial fixation is more problematic than femoral fixation in anterior cruciate ligament reconstruction. HYPOTHESIS: There is no difference in initial fixation strength among hamstring tendon graft tibial fixation devices. STUDY DESIGN: Randomized experimental study. METHODS: Each of six devices used to fix 120 quadrupled human semitendinosus-gracilis tendon grafts into porcine tibiae was tested 10 times with a single-cycle load-to-failure test and 10 times with a 1500-cycle loading test. Specimens surviving cyclic loading were subjected to a single-cycle load-to-failure test. RESULTS: Intrafix (1332 N) was the strongest in the single-cycle load-to-failure test, followed by WasherLoc (975 N), tandem spiked washer (769 N), SmartScrew ACL (665 N), BioScrew (612 N), and SoftSilk (471 N). After cyclic-loading tests, Intrafix showed the lowest residual displacement (1.5 mm) and was also strongest (1309 N) in the single-cycle load-to failure test after the cyclic-loading test, followed by WasherLoc (3.2 mm; 917 N). CONCLUSION: The Intrafix provided clearly superior strength in the fixation of hamstring tendon grafts to the tibial drill hole. CLINICAL RELEVANCE: Some caution may be warranted when using the implants that showed increased residual displacement, especially if aggressive rehabilitation is to be used. Preconditioning of the hamstring tendon graft-implant complex before tibial fixation is needed. PMID- 12642251 TI - Iontophoretic administration of dexamethasone sodium phosphate for acute epicondylitis. A randomized, double-blinded, placebo-controlled study. AB - BACKGROUND: A better treatment modality is needed to control the pain of medial or lateral epicondylitis (tennis elbow). HYPOTHESIS: Dermal iontophoretic administration of dexamethasone sodium phosphate will be significantly more effective in controlling pain than a placebo in patients with medial or lateral elbow epicondylitis. STUDY DESIGN: Randomized, double-blinded, placebo-controlled study. METHODS: On six occasions, 1 to 3 days apart within 15 days, 199 patients with elbow epicondylitis received 40 mA-minutes of either active or placebo treatment. RESULTS: Dexamethasone produced a significant 23-mm improvement on the 100-mm patient visual analog scale ratings, compared with 14 mm for placebo at 2 days and 24 mm compared with 19 mm at 1 month. More patients treated with dexamethasone than those treated with placebo scored moderate or better on the investigator's global improvement scale (52% versus 33%) at 2 days, but the difference was not significant at 1 month (54% versus 49%). Investigator-rated pain and tenderness scores favored dexamethasone over placebo at 2 days. Patients completing six treatments in 10 days or less had better results than those treated over a longer period. CONCLUSIONS: Iontophoresis treatment was well tolerated by most patients and was effective in reducing symptoms of epicondylitis at short-term follow-up. PMID- 12642252 TI - Anatomic reconstruction of the posterior cruciate ligament after multiligament knee injuries. A combination of the tibial-inlay and two-femoral-tunnel techniques. AB - BACKGROUND: Neither operative nor nonoperative treatment of posterior cruciate ligament rupture after multiligament knee injuries have shown very favorable outcomes. HYPOTHESIS: Reconstruction of the posterior cruciate ligament by combining the tibial-inlay and two-femoral-tunnel techniques will result in improved stability and functional outcomes. STUDY DESIGN: Prospective cohort study. METHODS: Twenty-nine patients with 30 posterior cruciate ligament ruptures and multiligament knee injuries treated with the combined technique were evaluated with clinical, radiographic, and functional outcome measures. RESULTS: All patients had a clinical examination result indicating joint stability (0 or 1+) at an average follow-up of 25 months (range, 15 to 39). Twenty-three knees had no laxity, and seven had 1+ laxity. The KT-2000 arthrometer data documented less than 0.5 mm of side-to-side mean difference for both posterior displacement and total anterior-posterior displacement at both 30 degrees and 70 degrees of knee flexion. Knee range of motion was a mean extension of 1 degrees (range, 0 degrees to 10 degrees ) and a mean flexion of 124 degrees (range, 75 degrees to 145 degrees ). Mean Lysholm knee score was 89.4. CONCLUSIONS: Reconstruction with a combination tibial-inlay and two-femoral-tunnel technique provides good results after multiligament knee injuries. All patients had a stable posterior cruciate ligament at most recent clinical follow-up, and 77% had no laxity at all. PMID- 12642253 TI - Arthroscopic repair for traumatic posterior shoulder instability. AB - BACKGROUND: The role of arthroscopic repair in the treatment of posterior shoulder instability remains poorly defined. PURPOSE: To evaluate the results of arthroscopic repair of posterior Bankart lesions. STUDY DESIGN: Retrospective review. METHODS: Records were reviewed of 27 shoulders (26 patients). All of the patients were male with a mean age of 28.7 years; in all cases symptoms were preceded by a traumatic event. Fourteen of the patients had 2+ to 3+ posterior translation noted under preoperative anesthesia. The posterior capsulolabral complex was found to be detached from the glenoid rim in all cases; bioabsorbable tack fixation was used for repair. RESULTS: At a mean follow-up of 5.1 years, no patients demonstrated a range of motion deficit. Muscle weakness (grade 4/5) in external rotation was noted in two patients (8%). There was no instability greater than 1+ in the anterior, posterior, or inferior directions. The mean L'Insalata shoulder score was 90.0 +/- 13.9. The mean SF-36 physical and mental component scores were 50.4 +/- 7 and 53.9 +/- 9, respectively. Symptoms of pain and instability were eliminated in 24 patients (92%). Two patients (8%) required additional surgery after arthroscopic repair of the posterior Bankart lesion. Radiographs demonstrated that there had been no progressive glenohumeral joint degeneration. CONCLUSIONS: Arthroscopic repair of the posterior capsulolabral complex is an effective means of eliminating symptoms of pain and instability associated with posterior Bankart lesions of traumatic origin. PMID- 12642254 TI - Lack of hormonal influences on mechanical properties of sheep knee ligaments. AB - BACKGROUND: The number of anterior cruciate ligament injuries in female athletes exceeds that in male athletes at similar competitive levels. This difference has been attributed by some authors to hormone-mediated alteration in knee laxity in women. HYPOTHESIS: Sheep anterior cruciate and medial collateral ligament strength and stiffness are not altered by administration for 6 months of estrogen or a selective estrogen receptor agonist (raloxifene). STUDY DESIGN: Controlled laboratory study. METHODS: Thirty-eight mature ewes were divided into five groups: sham operation (N = 6), ovariectomy (N = 9), ovariectomy and estradiol implant (N = 7), low-dose raloxifene (N = 9), and high-dose raloxifene (N = 7). After 6 months, the animals were sacrificed and ligaments were tested along with those from five rams' knees. RESULTS: No differences were found between treatment groups for maximum force, stiffness, energy to failure, or failure site. The ultimate stress of the rams' anterior cruciate ligaments was significantly higher than that of the ewes. CONCLUSIONS: Estrogen and estrogen receptor agonists at physiologic levels do not lead to decreased knee ligament strength. CLINICAL RELEVANCE: The female hormonal milieu may not be responsible for the increased incidence of anterior cruciate ligament injury in female athletes compared with their male counterparts. PMID- 12642255 TI - Repair of locked bucket-handle meniscal tears in knees with chronic anterior cruciate ligament deficiency. AB - BACKGROUND: Large bucket-handle meniscal tears in knees with chronic anterior cruciate ligament deficiency have been considered avascular and, thus, irreparable. HYPOTHESIS: The rate of meniscal healing is higher than previously reported. STUDY DESIGN: Prospective cohort study. METHODS: Fifty-nine patients underwent staged surgical procedures of meniscal repair followed at an average of 77 +/- 58 days by ligament reconstruction once full range of motion was obtained. RESULTS: Fifty-two patients with 55 meniscal repairs were available for follow up. At reconstruction, 30 menisci (55%) appeared healed; 19 (34%), partially healed; and 6 (11%) showed no healing (only 4 were removed). Of 43 tears in the white-on-white zone, 21 appeared healed; 17, partially healed; and 5 showed no healing. Of 11 in the red-on-white zone, 8 appeared healed; 2, partially healed; and 1 showed no healing. One meniscal tear in the red-on-red zone appeared healed. At an average follow-up of 4.3 +/- 3.1 years, 36 of the 43 (83.7%) white on-white meniscal repairs remained asymptomatic; all repairs in the other zones remained asymptomatic. CONCLUSION: Locked bucket-handle meniscal tears heal at a high rate when repaired as an isolated procedure, even when full weightbearing and activity before reconstruction is allowed and when the tear is in the white on-white zone. PMID- 12642256 TI - Codominance of the individual posterior cruciate ligament bundles. An analysis of bundle lengths and orientation. AB - BACKGROUND: It is unclear how each bundle of the posterior cruciate ligament contributes to posterior knee stability. HYPOTHESIS: Changes in bundle orientation and length occur such that neither bundle dominates in restraining posterior tibial motion throughout knee flexion and extension. STUDY DESIGN: Controlled laboratory study. METHODS: Six fresh-frozen cadaveric knees were studied in a joint-testing rig with individual quadriceps and hamstring muscle loading. Kinematic data for the tibia and femur were obtained at knee flexion angles from 0 degrees to 120 degrees. The joint was then disarticulated, and the insertions of the two bundles on the tibia and femur were digitized. RESULTS: Length of the anterolateral bundle increased with increasing knee flexion angle from 10 degrees to 120 degrees. Length of the posteromedial bundle decreased with increasing knee flexion angle from 0 degrees to 45 degrees and increased slightly from 60 degrees to 120 degrees. Length of the anteromedial bundle was significantly less than that of the posteromedial at 0 degrees, 10 degrees, and 20 degrees of knee flexion. The anterolateral bundle was significantly more horizontal at flexion angles of 0 degrees, 10 degrees, 20 degrees, 30 degrees, and 45 degrees (P < 0.05). The posteromedial bundle was more horizontal at 120 degrees. CONCLUSIONS: Changes in orientation take place such that neither bundle dominates in restraining posterior tibial motion throughout knee flexion and extension. CLINICAL RELEVANCE: Double-bundle reconstructions achieve more physiologic knee function. PMID- 12642257 TI - Anterior drawer test for acute anterior talofibular ligament injuries of the ankle. How much load should be applied during the test? AB - BACKGROUND: There is a lack of consensus regarding the magnitude of load for performing the anterior drawer test in evaluating acute ankle injuries. PURPOSE: To determine how much load should be applied during the anterior drawer test to detect the integrity of the anterior talofibular ligament. METHODS: First, the anterior-posterior load-displacement response of nine cadaveric ankles was measured. Second, anterior displacement of the ankle was measured at 30 and 60 N of anterior load in 14 patients with acute tears of the anterior talofibular ligament. RESULTS: In the cadaver study, the increased displacement by sectioning of the ligament measured at 10, 20, 30, and 40 N of anterior load were significantly greater than those measured at 60 N. In vivo examination of the subjects without anesthesia demonstrated that the injured-to-normal displacement value at 30 N of anterior load was significantly greater than the value at 60 N. CONCLUSIONS: This study suggests that a large magnitude of anterior load is not necessary to detect the integrity of the ligament during the anterior drawer test. CLINICAL RELEVANCE: When evaluating the integrity of the anterior talofibular ligament in cases of acute ankle ligament injury, a relatively low magnitude load should be applied. PMID- 12642258 TI - Jump landing strategies in male and female college athletes and the implications of such strategies for anterior cruciate ligament injury. AB - BACKGROUND: Female athletes are more likely than male athletes to injure the anterior cruciate ligament. Causes of this increased injury incidence in female athletes remain unclear, despite numerous investigations. HYPOTHESIS: Female athletes will exhibit lower hamstring muscle activation and smaller knee flexion angles than male athletes during jump landings, especially when the knee muscles are fatigued. STUDY DESIGN: Controlled laboratory study. METHODS: Eight female and six male varsity college basketball athletes with no history of knee ligament injury performed jump landings on the dominant leg from a maximum height jump and from 25.4 cm and 50.8 cm high platforms under nonfatigued and fatigued conditions. Knee joint angle and surface electromyographic signals from the quadriceps, hamstring, and gastrocnemius muscles were recorded. RESULTS: Women landed with greater knee flexion angles and greater knee flexion accelerations than men. Knee muscle activation patterns were generally similar in men and women. CONCLUSION: As compared with male college basketball players, female college basketball players did not exhibit altered knee muscle coordination characteristics that would predispose them to anterior cruciate ligament injury when landing from jumps. This conclusion is made within the parameters of this study and based on the observation that hamstring muscle activation was similar for both groups. The greater knee flexion we observed in the female subjects would be expected to decrease their risk of injury. CLINICAL RELEVANCE: Factors other than those evaluated in this study need to be considered when attempting to determine the reasons underlying the increased incidence of anterior cruciate ligament injuries consistently observed in elite female athletes. PMID- 12642259 TI - The soleus muscle acts as an agonist for the anterior cruciate ligament. An in vitro experimental study. AB - BACKGROUND: Although the quadriceps muscles are known antagonists for the anterior cruciate ligament and the hamstring muscles are known agonists, the influence of the calf muscles on knee stability is not well understood. HYPOTHESIS: The soleus muscle acts as an anterior cruciate ligament agonist and the gastrocnemius muscle acts as an anterior cruciate ligament antagonist. STUDY DESIGN: Controlled laboratory study. METHODS: Six cadaveric knees were tested with individual and combined activation of the gastrocnemius and soleus muscles to determine the influence of simulated muscle contraction on tibiofemoral motion. RESULTS: At all flexion angles, applying the soleus muscle force tended to translate the tibia posteriorly, whereas applying the gastrocnemius muscle force tended to translate the tibia anteriorly. Applying the soleus and gastrocnemius muscle forces together also tended to translate the tibia anteriorly. The average anterior and posterior tibial translations were greatest at 50 degrees of flexion. CONCLUSIONS: The soleus muscle is capable of acting as an agonist for the anterior cruciate ligament and the gastrocnemius muscle can act as an antagonist. CLINICAL RELEVANCE: A better understanding of the agonistic behavior of the soleus muscle on the anterior cruciate ligament may lead to the development of training and rehabilitation strategies that could reduce the incidence of injury and improve function in both patients with anterior cruciate ligament deficiency and patients who have undergone anterior cruciate ligament reconstruction. PMID- 12642260 TI - Anteromedial capsular redundancy and labral deficiency in shoulder instability. AB - BACKGROUND: Redundancy of the anteromedial capsule of the shoulder may persist despite proper tensioning of the capsule and repair of a Bankart lesion during an anteroinferior capsular shift procedure. HYPOTHESIS: A barrel-stitch suture technique incorporated into a capsular shift procedure is effective in achieving satisfactory shoulder stability. STUDY DESIGN: Uncontrolled retrospective review. METHODS: A barrel-stitch technique was used for patients identified as having anteromedial capsular redundancy during a capsular shift procedure for anteroinferior instability. RESULTS: The incidence of anteromedial capsular redundancy and labral deficiency was 49% (38 of 78). Patients with anteromedial capsular redundancy had a significantly greater number of dislocations before surgery (16.1 +/- 21.3 versus 7.4 +/- 7.4) and a greater duration of symptoms (79.8 +/- 84.2 versus 31.6 +/- 32.2 months). The mean postoperative Rowe score of patients with anteromedial capsular redundancy was 88.7 +/- 14.8, with 92% having excellent or good results, compared with 88.9 +/- 14.8 in the remaining patients and 93% excellent or good results. CONCLUSIONS: Anteromedial capsular redundancy is associated with longer preoperative duration of symptoms and more dislocations, but effective treatment can be achieved with a capsular shift procedure augmented with medial capsular imbrication with a barrel stitch. PMID- 12642261 TI - Radiofrequency thermal effects on the human meniscus. An in vitro study of systems with monopolar and bipolar electrodes. AB - BACKGROUND: No data exist on the cutting efficiency of monopolar versus bipolar radiofrequency energy application systems on human meniscal tissue. PURPOSE: To compare the effects of monopolar and bipolar thermal energy systems on human meniscal tissue. STUDY DESIGN: Controlled laboratory study. METHODS: Fresh-frozen menisci were cut in cross-section into 180 pie-shaped specimens. A specially designed jig was used to consistently apply radiofrequency energy to the tissue under a constant 30-g force. Three different systems were tested at the low, middle, and high ranges, with application times of 1 and 3 seconds. Thermal effects were measured by image analysis microscopy. RESULTS: No significant differences in thermal effects were found with respect to energy output for each system. Both the individual system tested and the application time had statistically significant effects on thermal damage, with the individual system tested having a greater effect. The mean depths of thermal change produced by the Mitek (bipolar) device were 564 and 648 microm at 1 and 3 seconds applications, respectively. The Arthrocare device (bipolar) produced depths of 1444 and 1697 microm at 1 and 3 seconds. The Oratec device (monopolar) produced depths of 895 and 1057 microm, respectively. CONCLUSIONS: A differential thermal effect was created in the meniscal tissue by three commercially available radiofrequency systems. Within the parameters of the experiment, all three systems limited thermal damage to a depth of less than 2 mm. The results appeared to depend more on the particular system used, not whether it had monopolar or bipolar electrodes. CLINICAL RELEVANCE: These data imply reasonably safe (less than 2 mm) thermal changes in the meniscus after radiofrequency energy application from these three systems. PMID- 12642262 TI - Anatomy of the superior glenoid rim. Repair of superior labral anterior to posterior tears. AB - BACKGROUND: Successful placement of a fixation device on the superior glenoid rim during superior labrum repairs requires accurate knowledge of the glenoid rim anatomy. PURPOSE: To investigate the normal bony anatomy of the superior glenoid rim. STUDY DESIGN: Descriptive anatomic study. METHODS: Twenty cadaveric glenoid specimens were scanned to obtain cross-sectional images with peripheral quantitative computed tomography in three different positions, each perpendicular to the articular surface. Two straight lines were drawn along the interior bony margins of the articular surface and cortex, and image analysis software was used to calculate the angle between these lines. Three bony angles were measured. RESULTS: The bony angles from the 10:30-, 12-, and 1:30-o'clock cross-sections were 55 degrees +/- 5 degrees, 64 degrees +/- 5 degrees, and 62 degrees +/- 8 degrees, respectively. The posterosuperior angle (at the 10:30-o'clock position) was statistically significantly lower than the superior and anterosuperior angles. Intraobserver variation was less than 3%. CONCLUSIONS: The most superior point of the glenoid rim (12-o'clock position) seems to provide the most bone stock for anchor insertion. The available bone support was found to decrease posteriorly on the glenoid rim. CLINICAL RELEVANCE: During superior labral repairs, the anchor or fixation device should be inserted at approximately a 30 degrees angle in relation to the articular surface for maximal bone support. PMID- 12642263 TI - Acute grade III medial collateral ligament injury of the knee associated with anterior cruciate ligament tear. The usefulness of magnetic resonance imaging in determining a treatment regimen. AB - BACKGROUND: The appropriate management of acute grade III medial collateral ligament injury when it is combined with a torn anterior cruciate ligament has not been determined. HYPOTHESIS: Magnetic resonance imaging grading of grade III medial collateral ligament injury in patients who also have anterior cruciate ligament injury correlates with the outcome of their nonoperative treatment. STUDY DESIGN: Prospective cohort study. METHODS: Seventeen patients were first treated nonoperatively with bracing. Eleven patients with restored valgus stability received anterior cruciate ligament reconstruction only, and six with residual valgus laxity also received medial collateral ligament surgery. RESULTS: Magnetic resonance imaging depicted complete disruption of the superficial layer of the medial collateral ligament in all 17 patients and disruption of the deep layer in 14. Restoration of valgus stability was significantly correlated with the location of superficial fiber damage. Damage was evident over the whole length of the superficial layer in five patients, and all five patients had residual valgus laxity despite bracing. Both groups had good-to-excellent results 5 years later. CONCLUSIONS: Location of injury in the superficial layer may be useful in predicting the outcome of nonoperative treatment for acute grade III medial collateral ligament lesions combined with anterior cruciate ligament injury. PMID- 12642264 TI - Shock wave application for chronic plantar fasciitis in running athletes. A prospective, randomized, placebo-controlled trial. AB - BACKGROUND: Recent articles have reported success with repeated low-energy shock wave application for treatment of chronic plantar fasciitis in runners. HYPOTHESIS: Shock wave treatment for chronic plantar fasciitis is safe and effective. STUDY DESIGN: Prospective, randomized, placebo-controlled trial. METHODS: Forty-five running athletes with intractable plantar heel pain for more than 12 months were enrolled; half were assigned to a treatment group that received three applications of 2100 impulses of low-energy shock waves, and half received sham treatment. Follow-up examinations were performed at 6 months and at 1 year by a blinded observer. RESULTS: After 6 months, self-assessment of pain on first walking in the morning was significantly reduced from an average of 6.9 to 2.1 points on a visual analog scale in the treatment group and from an average of 7.0 to 4.7 points in the sham group. The mean difference between groups was 2.6 points. After 12 months, there was a further reduction of pain in both groups, to an average 1.5 points in the treatment group, and to 4.4 points in the sham group. CONCLUSION: Three treatments with 2100 impulses of low-energy shock waves were a safe and effective method for treatment of chronic plantar fasciitis in long-distance runners. PMID- 12642265 TI - Risk factors for injury in middle school football players. AB - BACKGROUND: Little is known about the frequency of or risk factors for injuries in middle school or junior high school football players. PURPOSE: To examine the associations of player characteristics (injury history, conditioning, player position, special equipment) and physical parameters (body mass index, weight, height, grip strength) with risk of injury. STUDY DESIGN: Prospective cohort study. METHODS: We documented risk factors for injury in 646 middle school football players, 10 to 15 years of age, in the Oklahoma City, Oklahoma, school district during the 1998 and 1999 seasons. Player characteristics and physical parameters were measured at the beginning of both seasons. Logistic regression methods were used to determine whether baseline variables were associated with the odds of subsequent injury. RESULTS: More playing experience was the only variable significantly associated with the risk of injury in multivariate analyses. This association was observed regardless of the type of injury and even after indirectly controlling for time at risk of injury by restricting analyses to first-string players. Increasing age was significantly associated with the risk of fractures. CONCLUSIONS: Results suggest that physical characteristics play a minor role in risk of injury from football in this age group. PMID- 12642267 TI - Severe quadriceps muscle contusions in athletes. A report of three cases. PMID- 12642266 TI - Comparison of three techniques of anterior cruciate ligament reconstruction with bone-patellar tendon-bone graft. Differences in anterior tibial translation and tunnel enlargement with each technique. AB - BACKGROUND: Anatomic fixation of the graft in anterior cruciate ligament reconstruction has improved stability of the reconstructed knee joint in experimental studies. HYPOTHESIS: Anatomic fixation of the bone-patellar tendon bone autograft will reduce tunnel enlargement and improve clinical results. STUDY DESIGN: Prospective cohort study. METHODS: Sixty patients were randomly divided into three groups: a nonanatomic fixation group (traditional single-incision reconstruction), an anatomic fixation group (reconstruction in which a bone plug was grafted into the tibial tunnel), and an anatomic fixation group with all inside reconstruction. Stability of the knee joint was examined with a KT-1000 arthrometer at 2, 4, 6, 12, and 24 months after surgery. At 12 months, anteroposterior and lateral radiographs were made to assess tunnel enlargement. RESULTS: Although the magnitude of tibial displacement gradually increased after reconstruction in all three groups, the anatomic fixation group had significantly better stability than the groups undergoing nonanatomic fixation or all-inside anatomic fixation at 4 and 6 months after anterior cruciate ligament reconstruction. However, there was no significant difference between the three groups at 24 months. Regarding the tibial tunnel, residual rates and enlargement of tunnels were different between the reconstruction techniques. The nonanatomic reconstruction group had significantly greater tunnel enlargement. CONCLUSION: Anatomic fixation of the graft decreased the tunnel enlargement but had no effect on knee stability at 2-year follow-up. PMID- 12642268 TI - Chronic ischial apophysitis in a gymnast treated with transapophyseal drilling to effect "apophysiodesis". A case report. PMID- 12642269 TI - Solitary intraarticular lipoma causing sudden locking of the knee: a case report and review of the literature. PMID- 12642270 TI - A review of the special tests associated with shoulder examination. Part II: laxity, instability, and superior labral anterior and posterior (SLAP) lesions. AB - This is the second of a two-part article describing the various tests that have been used to examine the shoulder to find and treat problems in that area. Part I of this article (January/February 2003, pages 154-160) focused on tests used to examine rotator cuff abnormalities. This article attempts to clarify the tests of laxity, instability, and the superior labral anterior and posterior (SLAP) lesions by presenting them as described by the original authors, with the additional aim of providing a source for those wishing to refresh their knowledge without the need to refer to the original source material. PMID- 12642271 TI - Current concepts in arthroscopic rotator cuff repair. AB - The interest in arthroscopic rotator cuff repair has increased exponentially over the last 5 years. Although the operative technique of repair continues to evolve, there are now several studies reporting excellent results after arthroscopic repair of rotator cuff tears. In this review, we focus on new concepts and techniques related to arthroscopic rotator cuff repair that have been recently introduced. PMID- 12642272 TI - Fatigue testing of some orthopaedic fixation devices. PMID- 12642273 TI - The partial-thickness rotator cuff tear: is acromioplasty without repair sufficient? PMID- 12642274 TI - The treatment of resistant, painful tendinopathies results in frustration for athletes and health professionals alike. PMID- 12642275 TI - Demonstration of renal arterial anatomy and tumour neovascularity for vascular mapping of renal cell carcinoma: the value of CO2 angiography. AB - The aim of this study was to compare the efficacy of CO(2) angiography with that of iodinated contrast angiography for vascular mapping prior to partial nephrectomy for presumed renal cell carcinoma. 13 consecutive patients were studied and all patients underwent selective renal angiography using both CO(2) and iodinated contrast medium. Digitally subtracted images were acquired and compared. Seven male and six female patients, with a median age of 58 years (range 46-74 years), were examined. On comparing images the main renal artery was visualized in all cases with both contrast agents. The segmental vessels were seen in 7 of 13 CO(2) studies and 12 of 13 iodinated contrast studies. CO(2) was also inferior in the depiction of tumour circulation, showing it in 4 of 13 cases compared with 9 of 13 cases using iodinated contrast. It therefore appears that CO(2) angiography offers no diagnostic advantage and is also inferior to iodinated contrast angiography in the pre-operative vascular mapping of renal tumours. PMID- 12642276 TI - Viewing conditions for diagnostic images in three major Dublin hospitals: a comparison with WHO and CEC recommendations. AB - Accurate interpretation of X-ray images is dependent on image viewing conditions. A recent study in Ireland demonstrated that even with the advent of digital departments the majority of images are still viewed using viewing boxes. This investigation aimed to measure average viewing box brightness, percentage uniformity and ambient light levels in radiology and radiographer viewing areas and wards within three major Dublin hospitals. The results were compared with published recommendations by the World Health Organization and Commission of the European Communities. Following analysis of more than 4650 measurements, it was shown that mean values for average viewing box brightness for all departments failed to achieve recommended levels. Only one third of areas met the most lenient guideline for percentage uniformity. Ambient lighting was shown to be unacceptable for ward areas. For all three parameters, radiology areas generally fared best, with wards gaining the poorest scores. Following a 3 min cleaning regimen of viewing boxes, the average brightness and percentage uniformity were improved in 100% and 80%, respectively, of viewing boxes, but average brightness values remained below recommended levels. The importance of comprehensive quality assurance programmes for viewing boxes has been highlighted so that visualization of images is not reduced to sub-optimal levels. The need for consistent and more informative recommendations has been emphasized. PMID- 12642277 TI - Motor functional MRI for pre-operative and intraoperative neurosurgical guidance. AB - Functional MRI (fMRI) may provide a means of locating areas of eloquent cortex that can be used to guide neurosurgeons in their quest to maximize intracerebral tumour resection whilst minimizing post-procedural neurological deficits. This work aimed to develop and provide an initial assessment of such a technique. 19 patients with mass lesions close to the primary motor cortex underwent fMRI at 1.5T. A single shot echo planar technique was used to acquire data corresponding to right and left hand movement. Resultant activation maps were used to aid pre surgical planning. Data was used in conjunction with an intraoperative navigation system in 13 cases. Activation was attributed to primary motor, primary somatosensory or supplementary motor cortex in 17 of 19 subjects. No permanent changes in motor deficit were detected post surgery. The additional information provided by fMRI, particularly when incorporated into a neuronavigation guided craniotomy, was deemed highly valuable to the neurosurgeon as it enabled safe resection of tumour in anatomical locations previously deemed to be too high risk for safe resection using conventional (non-fMRI-guided) technique. This observation is reinforced by the fact that no patients suffered permanent neurological deficit after radical tumour debulking (surgical estimates >90% tumour resection). PMID- 12642278 TI - Comparative analyses of the dynamic properties of the rectum studied by cryo sections of human cadavers and pelvic CT scans of patients. AB - Optimization of radiotherapy treatment plans based on dose-volume histograms relies on accurate organ delineation. Hollow organs, such as the rectum, are difficult and time-consuming to delineate owing to unclear visualization of the border between wall tissue and filling. Automated hollow organ delineation would be a valuable tool, but its development depends upon improved understanding of the dynamics of the rectum in response to filling. Two reasonable assumptions proposed in the literature are that (1) the rectal wall tissue along a constant length of the rectal cylinder is preserved over time and (2) the rectal wall tissue is distributed homogeneously along the cylinder. Therefore, variations in wall thickness can be explained by variable rectal filling. To investigate these assumptions, transversal cross-sectional areas enclosed by the outer contour (A(out)) and inner contour (A(in)) of the rectum were recorded from digital photographs of cadaver cryo-sections from the U.S. National Library of Medicine's Visible Human Project. In addition, A(out) and A(in) were recorded from 19 CT scans of 5 of our own patients. The transversal cross-sectional area of the wall of the rectum, A(wall)=A(out)-A(in), was calculated. The data derived both from cryo-sections and repetitive CT scans of patients, revealed that there was a significant correlation between A(wall) and A(out), in contradiction to assumption (1) stated above (male cryo-sections: p<0.001, female cryo-sections: p=0.03, repetitive CT scans p<0.001). Moreover, the mean A(wall) calculated from one CT scan differed significantly from the mean A(wall) from other CT scans and was correlated with the mean A(out), i.e. rectal filling (p<0.001). This finding was confirmed by careful analysis of another study (p=0.001) and opposes assumption (2). Hence, the amount of wall tissue within a constant length of rectum is not preserved over time, but increases with increased filling. This implies that the longitudinal length of the rectum decreases in response to distension of the organ. PMID- 12642279 TI - Dose dependence of the growth rate of multicellular tumour spheroids after irradiation. AB - The present study investigated differences in the growth rate of multicellular tumour spheroids of the MCF-7 line of human breast cancer before and after their irradiation. Growth of the spheroids was analysed according to a model based on a Gompertz function. In this model, normalization to a common initial volume is achieved in a way that enables meaningful comparisons to be made between the results obtained for each spheroid. For irradiated spheroids the model includes an additional term to take account of sterilized cells. We found that the growth rate observed before irradiation is not fully recovered by irradiated spheroids and that growth recovery reduces with higher irradiation doses. Surviving fractions obtained at doses below 3 Gy are comparable with those found in clonogenic assays on spheroids of the same cellular line. At larger doses, discrepancies between the different studies are considerable. PMID- 12642280 TI - Quantitative assessment of the radiation hazards and risks in sentinel node procedures. AB - Sentinel node localization using an injected radiopharmaceutical and a gamma probe is performed in many hospitals. Employers have a duty to give appropriate training to staff who may not have been previously formally trained to work with unsealed radioactive sources. A study was performed to assess hazards and risks at all stages of the localization procedure. Whole body doses and finger doses of imaging, surgery and pathology staff were determined. The activity remaining in the tumour specimen, excised nodes and disposable waste from the operating theatre was measured. Any radioactive contamination of the operating theatre and equipment was also ascertained. All results were then assessed in light of current UK radiation protection legislation for the protection of staff and members of the public. Results showed that radiation doses are low and no additional procedures are required for protection of staff, provided the usual procedures for biohazards are in place. However, an information sheet has been prepared for the reassurance of staff, and theatre swabs may need to be stored temporarily before disposal. Injecting and imaging on the day before surgery is preferred, compared with injecting and imaging before surgery on the same day, since this gives lower radiation doses to staff, lower activity in excised specimens and waste, and provides a higher count rate giving better image quality. PMID- 12642281 TI - Evaluation of calcaneus bone densitometry against hip and spine for diagnosis of osteoporosis. AB - Dual X-ray absorptiometry (DXA) is now widely available as a method for assessing bone density. However, the place of peripheral bone densitometry in clinical practice for diagnosis of osteoporosis is not yet clear. To examine the potential use in our district general hospital setting, we compared calcaneus measurements with conventional DXA of the hip and spine in 100 patients referred for assessment following identification of risk factors for osteoporosis. Measurements were made on both heels and the results were found to be similar but not completely interchangeable. Use of receiver operating characteristic curves confirmed that a threshold T-score of -1.6 could be used to identify many of the high risk subjects. However, there was only moderate agreement between fracture risk classifications derived from heel T-scores, and diagnostic classification (osteoporosis/osteopenia/normal) derived from axial DXA. The specificity of heel measurements was high, but sensitivity was poorer. Heel measurements could therefore be valuable in some circumstances for finding patients for whom treatment of osteoporosis would be appropriate, such as in a population with a low prevalence of osteoporosis. They may also be of value in a population with a high prevalence of disease, particularly if there were no alternative means of bone densitometry. However, with an intermediate prevalence, the relatively high risk of false negative values would mean that false reassurance could be given to many of those classed as "low risk". This could be a major drawback in clinical practice if heel densitometry were used as the initial investigation and axial measurements were also available, since they would give conflicting results for a substantial proportion of these patients. PMID- 12642282 TI - Measurement of dose-width product in panoramic dental radiology. AB - The National Radiological Protection Board has recommended the introduction of dose-width product (DWP) for the measurement of patient dose in panoramic dental radiology. The present work describes a method for measuring DWP using a pencil ionization chamber. The technique was tested on five panoramic dental units; the reproducibility of the method was 5.7%. In order to test the method, DWP was also assessed using thermoluminescent dosemeters and film. The results obtained agreed within 8.6% with those obtained using the pencil ionization chamber method. The proposed method appears to be simple and precise. PMID- 12642283 TI - Case report: Spinal cord herniation into a mediastinal neurenteric cyst: CT and MRI findings. AB - Split notochord syndrome is a spectrum of congenital spinal malformations that develops due to an adhesion between endoderm and ectoderm causing the "splitting" of notochord. Neurenteric cyst is one of the components of split notochord syndrome. We report CT and MRI findings of an unusual case with thoracal spinal cord herniation into a mediastinal neurenteric cyst. PMID- 12642284 TI - Case report: Conservative management of an arteriovenous fistula of the inferior epigastric artery. AB - We present a case of pseudoaneurysm and arteriovenous fistula of the inferior epigastric artery secondary to the placement of a drain during a surgical intervention. We stress the utility of colour Doppler ultrasound and arteriography embolisation in diagnosis and treatment, respectively. PMID- 12642285 TI - CT of a thickened-wall gall bladder. AB - This pictorial article reviews the various clinical entities that may cause mural thickening of the gall bladder encountered on contrast enhanced CT. PMID- 12642286 TI - Case of the month: A young man with a lump on the head. PMID- 12642287 TI - Stabilization of the hydrophilic sphere of iobitridol. PMID- 12642290 TI - The SERI-ARVO Meeting and future challenges of ophthalmic research in Asia. PMID- 12642291 TI - Primary care in ophthalmology. PMID- 12642292 TI - Behcet's syndrome. PMID- 12642294 TI - Is medicolegal work a duty? It is. PMID- 12642295 TI - Is ophthalmology evidence based? A clinical audit of the emergency unit of a regional eye hospital. AB - AIM: To evaluate the proportion of interventions that are evidence based in the acute care unit of a regional eye hospital. METHODS: A prospective clinical audit was carried out at Hong Kong Eye Hospital in July 2002 to investigate the extent to which ophthalmic practices were evidence based. The major diagnosis and intervention provided were identified through chart review. A corresponding literature search using Medline and the Cochrane Library was performed to assess the degree to which each intervention was based on current, best evidence. Each diagnosis intervention pair was accordingly analysed and graded. The level of best, current evidence supporting each intervention was graded and analysed. RESULTS: A total of 274 consecutive consultation episodes were examined. 22 cases were excluded since no diagnosis or intervention was made during the consultation. 108 (42.9%) patient interventions were found to be based on evidence from systematic reviews, meta-analyses, or randomised controlled trials (RCT). Evidence from prospective or retrospective observational studies supported the interventions in 86 (34.1%) patients. In 58 (23.0%) cases, no evidence or opposing evidence was found regarding the intervention. The proportion of evidence based on RCT or systematic reviews was higher for surgical interventions compared with non-surgical interventions (p=0.007). The proportion of interventions based on RCT or systematic reviews was higher for specialist ophthalmologists than trainee ophthalmologists (p=0.021). CONCLUSION: This study demonstrated that the majority of interventions in the ophthalmic unit were evidence based and comparable to the experience of other specialties. PMID- 12642296 TI - Unilateral visual impairment and health related quality of life: the Blue Mountains Eye Study. AB - BACKGROUND: There has been considerable recent interest in the impact of unilateral visual impairment on functional status and wellbeing, particularly in relation to second eye cataract surgery. AIM: To determine if unilateral visual impairment has a measurable impact on health related quality of life (HRQOL) in an older community, as assessed by the generic, multidimensional 36 item short form health survey (SF-36). METHODS: All participants of the second cross sectional Blue Mountains Eye Study (n = 3508) were invited to attend comprehensive eye examinations and complete an SF-36 questionnaire. Unilateral visual impairment was defined as visual acuity (VA) <6/12 in the worse eye and > or =6/12 in the better eye. Mild visual impairment was defined as VA <6/12 but > or =6/24, moderate as VA <6/24 but > or =6/60, and severe (blindness) as VA <6/60 in the worse eye. Cases with amblyopia (n = 48) were excluded. RESULTS: Complete data were available for 3108 participants; 227 (7.3%) had unilateral visual impairment (148 mild, 29 moderate, 50 severe). Moderate to severe non-correctable unilateral impairment was associated with poorer SF-36 profiles. After adjusting for age and sex, this group had significantly poorer scores than the unimpaired group in three of eight domains (p<0.05); limitations as a result of physical problems, social function, limitations because of emotional problems, and in the mental component score. Mental domains were more affected than physical domains. Unilateral impairment from undercorrected refraction did not measurably affect HRQOL. CONCLUSIONS: Moderate to severe non-correctable unilateral visual impairment caused by eye diseases such as cataract had a measurable impact on HRQOL. PMID- 12642297 TI - Risk factors for conjunctival squamous cell neoplasia: a matched case-control study. AB - AIMS: To identify roles of human papillomavirus (HPV) infection and solar elastosis as the risk factors for conjunctival squamous cell neoplasia (CSCN). METHODS: 30 consecutive pathological specimens, ranging from conjunctival intraepithelial neoplasia, carcinoma in situ, to invasive squamous cell carcinoma were retrieved from tissue archives. 30 controls were disease free conjunctiva from age and sex matched patients undergoing extracapsular cataract extraction. Two masked pathologists studied haematoxylin and eosin stains on paraffin embedded conjunctival tissues. Elastic stain for solar elastosis was blindly interpreted in comparison with negative and positive controls. HPV infection was studied by polymerase chain reaction and dot hybridisation. RESULTS: The mean age of CSCN patients was 54.9 years. The male to female ratio was 1:1. Solar elastosis was seen in 53.3% of CSCN and in 3.3% of controls with an odds ratio of 16.0 (95% CI, 2.49 to 670.96; p value = 0.0003). HPV DNA were not detected in any of the specimens. CONCLUSION: Solar elastosis is much more frequently found in CSCN cases than in their matched controls and is a risk factor for CSCN. These data are insufficient to conclude that HPV infection is a risk factor for CSCN. PMID- 12642298 TI - Ocular biometry in occludable angles and angle closure glaucoma: a population based survey. AB - AIM: To compare ocular biometric values in a population based sample of eyes with occludable angles, angle closure glaucoma, and normal subjects. METHOD: 2850 subjects from a population based glaucoma prevalence study underwent complete ocular examination including indentation gonioscopy. Ocular biometry was performed in all subjects classified to have occludable angles (n = 143); angle closure glaucoma (n = 22), and a random subgroup of 419 normal subjects. Ocular biometry readings between the groups were compared and statistically analysed using "t," "z," and Mann-Whitney U tests. RESULTS: The mean age among subjects with occludable angles (54.43 (SD 9.53) years) and angle closure glaucoma (57.45 (8.5) years) was significantly higher (p<0.001) than normal subjects (49.95 (9.95) years). Axial length was shorter (p<0.001) in the occludable angle group (22.07 (0.69) mm) compared to the normal group (22.76 (0.78) mm). Anterior chamber depth (ACD) was shallower (p<0.001) among subjects with occludable angles (2.53 (0.26) mm) than normal subjects (3.00 (0.30) mm). Lens thickness (LT) was greater (p<0.001) in people with occludable angles (4.40 (0.53) mm) compared to normal subjects (4.31 (0.31) mm). No significant difference was noted in axial length, ACD (p = 0.451), and LT (p = 0.302) between angle closure glaucoma and occludable eyes. CONCLUSION: South Indian eyes with angle closure glaucoma and occludable angles seem to have significantly shorter axial lengths, shallower anterior chambers and greater lens thickness compared to the normal group. PMID- 12642300 TI - Confocal microscopy of Aspergillus fumigatus keratitis. AB - AIM: To use a confocal microscope to characterise the treated and untreated courses of fungal keratitis. METHODS: In the first experiment, Aspergillus fumigatus stromal keratitis was produced in both eyes of seven New Zealand white rabbits. In the second experiment, keratitis was induced in right eyes of 20 rabbits. Group 1 rabbits were treated with topical fluconazole, group 2 rabbits received oral fluconazole, and group 3 rabbits were used as controls. The rabbits were examined with a slit lamp and confocal microscope 2, 6, 10, 14, and 20 days after inoculation. The corneal cultures were taken on days 2, 14, and 20 and biopsies were taken on days 2 and 22. RESULTS: On days 14 and 22 confocal microscopy was more sensitive than culture technique in both treated and untreated animals, since not all cases of fungal keratitis can be cultured. CONCLUSION: This study indicates that confocal microscopy is a rapid and sensitive diagnostic tool for both the early diagnosis and non-invasive follow up of fungal keratitis PMID- 12642301 TI - Pulverulent cataract with variably associated microcornea and iris coloboma in a MAF mutation family. AB - AIMS: To report the detailed clinical findings in a three generation pedigree with autosomal dominant cataract, microcornea, and coloboma resulting from mutation of the lens development gene, MAF. METHODS: Five members of a three generation pedigree with progressive cataracts underwent detailed ophthalmic examination to characterise associated ocular phenotypic features. RESULTS: The cataracts present in all affected individuals were cortical, and/or nuclear, pulverulent opacities. Corneal diameters of 10-10.25 mm were present in two family members. Axial lengths were in the normal range. Bilateral iris coloboma in the 6 o'clock position was present in one patient. Uveal melanoma was present in one patient, with uveal naevi in this and one other patient. CONCLUSION: The bZIP transcription factor MAF is a key lens development gene that regulates the expression of the crystallins. Individuals with a mutation in MAF may have pulverulent cataract alone or cataract in association with microcornea or iris coloboma. PMID- 12642299 TI - Associations of selected medications and visual function: the Beaver Dam Eye Study. AB - AIM: To investigate association of drug use and visual function. METHODS: A cross sectional population based study was carried out on participants in the 1993-5 examination phase of the Beaver Dam Eye Study. All drugs in current use by study participants were recorded. Performance based and self assessed visual functions were obtained at the time of the study evaluation. The main outcome measure was the relation of levels of visual functions by use of specific drugs. RESULTS: Many classes of drugs were associated with decreases in at least two performance based visual functions. For example, high blood pressure drugs were significantly associated with poorer best corrected visual acuity, poorer near vision, and poorer contrast sensitivity (p<0.001 for all). Patterns of association for self assessed visual functions were not as strong. However, use of glaucoma drops and benzodiazepines were associated with poorer self assessed visual functions in most circumstances cited. CONCLUSIONS: Many commonly used medications are inversely associated with visual functions in a middle and older aged population. This may influence the ability to perform complex tasks and quality of life. PMID- 12642302 TI - Comparison of algorithms for detection of localised nerve fibre layer defects using scanning laser polarimetry. AB - AIMS: To evaluate different algorithms used to analyse retinal nerve fibre layer thickness (RNFL) data obtained by scanning laser polarimetry, in order to compare their relative abilities to discriminate between patients with glaucomatous localised nerve fibre layer defects and normal subjects. METHODS: 48 eyes of 48 glaucomatous patients with localised RNFL defects and 53 eyes of 53 healthy subjects were included in this study. The localised RNFL defects were identified by RNFL photography and/or slit lamp biomicroscopic examination. All patients were submitted to RNFL examination using scanning laser polarimetry (GDx nerve fibre analyser, Laser Diagnostic Technologies, Inc, San Diego, CA, USA). Three methods of analysis of polarimetry data were used: GDx software provided parameters; RNFL thickness measurements in 16 equal sectors around the optic disc (sectoral analysis); and Fourier analysis of the curve of distribution of RNFL thickness measurements. Linear discriminant functions were developed to assess sensitivity and specificity of the sectoral based analysis and Fourier analysis and were compared to the GDx parameters. In addition, areas under the receiver operating characteristic (ROC) curves were compared. RESULTS: At a fixed specificity of 91%, the sensitivity of the linear discriminant function from sectoral data (LDF sectoral) was 81%, with an area under the ROC curve of 0.93. The linear discriminant function from Fourier measures had a comparable performance, with an area under the ROC curve of 0.93, and sensitivity of 71% for specificity at 91%. At the same specificity, the sensitivities of the GDx software provided parameters ranged from 15% to 40%. The areas under the ROC curves for the LDF sectoral and LDF Fourier were significantly greater than the ROC curve area for the single best GDx parameter. CONCLUSION: The sectoral based analysis and the Fourier analysis of RNFL polarimetry data resulted in an improved detection of eyes with glaucomatous localised nerve fibre layer defects compared to the GDx software provided parameters. PMID- 12642303 TI - Amiodarone induced optic neuropathy. AB - AIM: To determine the clinical features of amiodarone induced optic neuropathy, which may help distinguish it from non-arteritic anterior ischaemic optic neuropathy. METHODS: Retrospective observational case series of patients diagnosed with amiodarone induced optic neuropathy at the neuro-ophthalmology service from March 1998 to February 2001. Amiodarone was discontinued after discussion with the patient's cardiologist. Visual acuity, colour vision, automated perimetry, and funduscopy were performed on initial and follow up examinations. RESULTS: Three patients with amiodarone induced optic neuropathy presented with mildly decreased vision, visual field defects, and bilateral optic disc swelling. Upon discontinuing the medication, visual function and optic disc swelling slowly improved in all three patients. CONCLUSION: Amiodarone induced optic neuropathy can present with visual dysfunction, and is typically a bilateral process. Upon discontinuation of amiodarone, slow resolution of optic disc swelling occurs and visual function improves in some patients. PMID- 12642304 TI - Human recombinant interferon alfa-2a for the treatment of Behcet's disease with sight threatening posterior or panuveitis. AB - BACKGROUND: Behcet's disease is a multisystem vasculitis of unknown origin. Standard treatment mainly comprises systemic immunosuppressive agents. Ocular involvement, mostly posterior uveitis with retinal vasculitis, leads to blindness in 20-50% of the involved eyes within 5 years. The efficacy of interferon alfa-2a was studied in patients with sight threatening posterior uveitis or retinal vasculitis. METHODS: 50 patients were included in this open, non-randomised, uncontrolled prospective study. Recombinant human interferon alfa-2a (rhIFNalpha 2a) was applied at a dose of 6 million units subcutaneously daily. Dose reduction was performed according to a decision tree until discontinuation. Disease activity was evaluated every 2 weeks by the Behcet's disease activity scoring system and the uveitis scoring system. RESULTS: Response rate of the ocular manifestations was 92% (three non-responder, one incomplete response). Mean visual acuity rose significantly from 0.56 to 0.84 at week 24 (p<0.0001). Posterior uveitis score of the affected eyes fell by 46% every week (p<0.001). Remission of retinal inflammation was achieved by week 24. Mean Behcet's disease activity score fell from 5.8 to 3.3 at week 24 and further to 2.8 at week 52. After a mean observation period of 36.4 months (range 12-72), 20 patients (40%) are off treatment and disease free for 7-58 months (mean 29.5). In the other patients maintenance IFN dosage is three million units three times weekly. CONCLUSIONS: rhIFNalpha-2a is effective in ocular Behcet's disease, leading to significant improvement of vision and complete remission of ocular vasculitis in the majority of the patients. PMID- 12642306 TI - A comparative bioavailability study of three conventional eye drops versus a single lyophilisate. AB - AIM: To study the ocular bioavailability of a triple dose, single application of sodium fluorescein to the human anterior segment from a novel drug delivery device. METHODS: In a randomised, open label study 22 healthy volunteers applied a single lyophilisate to one eye (+1 minute) and three conventional eye drops (+1, 16, 31 minutes) of fluorescein ophthalmic solution to the fellow eye. The fluorescein dose of the lyophilisate was 204 mg corresponding to three conventional, preservative-free eye drops of 40 ml fluorescein SE Thilo 0.17% (68 micro g each) (Alcon). Fluorophotometry was performed (Fluorotron Master II Ocumetrics, USA) before and +15, 30, 45, 60, 120, 180, 240, 300, 360, 420 minutes after application. The fluorescein concentrations of the corneal stroma and mid anterior chamber were analysed by paired t test. RESULTS: Cornea and anterior chamber mean values (ng/ml) were significantly higher (p<0.018, paired t test) in the lyophilisate group up to 7 hours after application with the exception of +45 minutes. The mean fluorescein bioavailability from the lyophilisate was up to 11 times higher in the cornea and up to 8.7 times higher in the anterior chamber compared with the three preservative-free eye drops. CONCLUSION: A triple dose was delivered to the human eye with a single lyophilisate application for the first time. A significantly better bioavailability was achieved in the cornea and anterior chamber for up to 7 hours by means of drug application with lyophilisates. The application of medications by means of the lyophilisate will improve the treatment of, for example, glaucoma, bacterial, viral and fungal infections, as well as dry eye syndrome. PMID- 12642305 TI - Effects of lamellar keratotomy on postkeratoplasty astigmatism. AB - AIM: To determine the changes in postkeratoplasty astigmatism induced by lamellar keratotomy. METHODS: A prospective, non-randomised comparative trial of patients undergoing a hinged lamellar corneal flap for treatment of significant astigmatism after penetrating keratoplasty. Uncorrected visual acuity, best corrected visual acuity, refraction, and corneal topography were assessed at 1 and 3 months after the lamellar keratotomy. RESULTS: 17 eyes in 16 patients (13 M, 3F) were included in the study (mean age 48.2 years; range 20-86 years). Six of 17 eyes (35.3%) changed more than 1 dioptre (D) in spherical equivalent by 3 months. Nine of 17 eyes (52.9%) changed more than 1 D in sphere by 3 months. 12 of 17 eyes (70.6%) changed more than 1 D in refractive cylinder. Seven patients of 15 (46.7%) changed more than 1 D in corneal power as measured topographically. Five of 17 eyes (29.4%) changed in refractive cylinder axis more than 15 degrees and this was similar to the change measured topographically of four of 15 eyes (26.7%). Vector analysis showed 60% of eyes had a surgically induced astigmatism (SIA) vector of more than 1 D, including a net corneal astigmatism decrease of more than 1 D in four eyes and increase of more than 1 D in two eyes at 3 months after surgery. Complications of the lamellar keratotomy included two partial buttonholes and one partial wound dehiscence. CONCLUSIONS: The creation of a lamellar flap alone can have significant effects on the astigmatism following penetrating keratoplasty. LASIK for correction of postkeratoplasty astigmatism may be more accurately performed as a two stage procedure rather than a single stage, after the corneal effects of the lamellar keratotomy have stabilised. PMID- 12642307 TI - Five year results of viscocanalostomy. AB - AIM: To prospectively study the success rate and complications of viscocanalostomy, a non-penetrating glaucoma surgery. METHODS: Prospective non randomised consecutive case series of 57 eyes (57 patients) with medically uncontrolled primary and secondary open angle glaucoma. Viscocanalostomy was performed on all participants with injection of viscoelastic in the surgically created ostia of Schlemm's canal as well as in the scleral bed, the superficial scleral flap was loosely sutured. Intraocular pressure, visual acuity, and number of goniopunctures were measured. RESULTS: The mean follow up period was 34.1 months. The mean preoperative intraocular pressure (IOP) was 24.6 mm Hg; while the mean postoperative IOP was 5.6 mm Hg at day 1 and 13.9 mm Hg at 36 month. Patients who achieved IOP below 21 mm Hg with or without medication were 90% at 60 months, complete success rate (IOP<21 mm Hg without medication) was 60% at 60 months. 21 patients (37%) needed Nd:YAG goniopuncture postoperatively to control raised IOP, mean time for goniopuncture application was 9.4 months, mean pre goniopuncture IOP was 20.4 mm Hg and mean postgoniopuncture IOP was 12.6 mm Hg (p <0.0001). CONCLUSION: Viscocanalostomy appears to be a promising modification of filtering surgery. PMID- 12642308 TI - Awareness of glaucoma, and health beliefs of patients suffering primary acute angle closure. AB - AIM: To evaluate the factors associated with lack of awareness of glaucoma and late presentation to the doctor in Singapore Chinese patients with acute angle closure (AAC) METHODS: A prospective, hospital based case series of 105 patients aged 35 years and above who presented with a first attack of AAC in a tertiary hospital in Singapore was conducted. A research assistant interviewed all patients face to face in clinic and recorded demographic factors, awareness of glaucoma, and subjective barriers to seeing a doctor. The time from onset of symptoms to presentation at the clinic was noted. RESULTS: Overall, 22.9% of patients had heard of glaucoma. The multivariate adjusted odds ratio (OR) of unawareness of glaucoma in older people (> 60 years) was 1.5 (95% confidence interval (CI) 0.5 to 4.6), 3.2 (95% CI 1.1 to 9.2) for adults who were not working, and 13.8 (95% CI 1.3 to 146.7) for patients who had less than a pre university education. A significant proportion (31.7%) of patients presented to the doctor 24 hours or more after symptoms occurred. In a multiple logistic regression model, the adjusted OR of late presentation was 8.5 (95% CI 1.04 to 69.5) if there was no car access, 5.0 (95% CI 1.0 to 24.6) if the patients spoke Chinese, and 3.3 (95% CI 0.9 to 11.9) if there was nobody to accompany to hospital. CONCLUSION: Glaucoma awareness among patients suffering AAC was not high. Lack of awareness was associated with increasing age, lack of formal education, and unemployment. A significant proportion of patients seek medical attention late. Risk factors for late presentation include lack of car access, nobody to accompany the patient, and speaking the Chinese language primarily. Health education programmes may help increase the knowledge and awareness of glaucoma. PMID- 12642309 TI - Five year risk of progression of primary angle closure suspects to primary angle closure: a population based study. AB - AIM: To report progression of primary angle closure suspects (PACS) to primary angle closure (PAC) at the 5 year follow up of a population based sample. METHODS: 82 of 118 PACS who could be contacted and 110 randomly selected normals from a population based survey in 1995 were invited for a follow up examination in 2000. Progression to PAC was based on the development of raised IOP or synechiae in a PACS. RESULTS: 50 of the 82 PACS contacted were examined. 11 (22%; 95% CI 9.8 to 34.2) developed PAC (seven synechial and four appositional); all were bilateral PACS. Two of 50 people previously diagnosed as PACS were reclassified as normal. One person among the 110 normals progressed to PAC. The relative risk of progression among PACS was 24 (95% CI 3.2 to 182.4). There was no significant difference in axial length, anterior chamber depth, or lens thickness between those who progressed and those who did not. None of the patients developed optic disc or field damage attributable to angle closure. One angle closure suspect was diagnosed to have normotensive glaucoma. CONCLUSION: In this population based study of PACS the 5 year incidence of PAC was 22%; none developed functional damage. Bilateral PACS was a clinical risk factor for progression. PMID- 12642310 TI - Correlation of a scanning laser derived oedema index and visual function following grid laser treatment for diabetic macular oedema. AB - AIM: To correlate change of an oedema index derived by scanning laser tomography with change of visual function in patients undergoing grid laser photocoagulation for clinically significant diabetic macular oedema (DMO). METHODS: The sample comprised 24 diabetic patients with retinal thickening within 500 micro m of the fovea. Inclusion criteria included a logMAR visual acuity of 0.25, or better. Patients were assessed twice before a single session of grid laser treatment and within 1 week of, and at 1, 2, 4, and 12 weeks after, treatment. At each visit, patients underwent logMAR visual acuity, conventional and short wavelength automated perimetry (SWAP), and scanning laser tomography. Each visual function parameter was correlated with the mean oedema index. The mean oedema index represented the z-profile signal width divided by the maximum reflectance intensity (arbitrary units). A Pearson correlation coefficient (Bonferroni corrected) was undertaken on the data set of each patient. RESULTS: 13 patients exhibited significant correlation of the mean oedema index and at least one measure of visual function for the 10 degrees x 10 degrees scan field while 10 patients correlated for the 20 degrees x 20 degrees scan field. Seven patients demonstrated correlation for both scan fields. Laser photocoagulation typically resulted in an immediate loss of perimetric sensitivity whereas the oedema index changed over a period of weeks. Localised oedema did not impact upon visual acuity or letter contrast sensitivity when situated extrafoveally. CONCLUSIONS: Correlation of change of the oedema index and of visual function following grid laser photocoagulation was not found in all patients. An absence of correlation can be explained by the localised distribution of DMO in this sample of patients, as well as by differences in the time course of change of the oedema index and visual function. The study has objectively documented change in the magnitude and distribution of DMO following grid laser treatment and has established the relation of this change to the change in visual function. PMID- 12642311 TI - Intravitreal triamcinolone acetonide for exudative age related macular degeneration. AB - AIM: To evaluate the effect of intravitreal triamcinolone acetonide on the visual acuity of patients with exudative age related macular degeneration, to assess the duration of a possible effect, and to evaluate clinical side effects of the treatment. METHODS: The study included 67 patients (71 eyes) who presented with exudative age related macular degeneration of predominantly or total occult type (n = 68) or classic type (n = 3), and who received once, or repeatedly, an intravitreal injection of 25 mg of crystalline triamcinolone acetonide. Mean follow up time was 7.46 (SD 3.54) months (range 3.1-19.57 months). RESULTS: Visual acuity increased significantly (p <0.001) from 0.16 (0.11) to a mean maximum of 0.23 (0.17). Postoperative visual acuity was highest 1-3 months after the injection. 47 (66.2%) eyes gained in maximal visual acuity and 11 (15.5%) eyes lost in visual acuity. Intraocular pressure increased significantly (p <0.001) from 15.1 (3.1) mm Hg at baseline to a maximal value of 23.0 (8.25) mm Hg. At the end of follow up, intraocular pressure again decreased significantly (p<0.001) to 16.8 (4.9) mm Hg. No cases of postoperative infectious endophthalmitis, rhegmatogenous retinal detachment, or proliferative vitreoretinopathy occurred. Owing to a decrease in visual acuity after an initial increase, six patients received a second intravitreal triamcinolone acetonide injection after which visual acuity increased again in three eyes. CONCLUSIONS: Intravitreal injection of 25 mg of crystalline triamcinolone acetonide merits further study for the treatment of exudative age related macular degeneration. PMID- 12642312 TI - Risk factors for age related maculopathy in a Japanese population: the Hisayama study. AB - AIMS: To examine the risk factors for age related maculopathy (ARM) in a sample Japanese population. METHODS: In 1998, a cross sectional community survey was conducted among residents of Hisayama. A total of 596 men and 886 women living in Hisayama, Japan, aged 50 years or older consented to participate in the study. Each participant underwent a comprehensive examination that included an ophthalmic examination. The presence of ARM was determined by grading from fundus examination by indirect ophthalmoscopy, slit lamp examination, and colour fundus photographs. Using these cross sectional data, logistic regression analyses were performed to determine the risk factors for ARM. The following 10 possible risk factors were used: age, cataract, hypertension (history), hypertension (history or examination), diabetes, hyperlipidaemia, current smoker, alcohol intake, BMI, and WBC. RESULTS: ARM was detected in 19.5% of men and 14.9% of women. Men were found to have a significantly higher prevalence of ARM than women. Multiple logistic regression analysis showed that age and hypertension (history or examination) were significantly associated with ARM in men, whereas only age was a significant risk factor for ARM in women. CONCLUSIONS: This study suggests that higher age and male sex are relevant risk factors for ARM in Japan. In addition, hypertension is a relevant risk factor in men. PMID- 12642313 TI - Progression of phenotype in Leber's congenital amaurosis with a mutation at the LCA5 locus. AB - BACKGROUND: Leber's congenital amaurosis (LCA) accounts for 5% of inherited retinal disease and is usually inherited as an autosomal recessive trait. Genetic and clinical heterogeneity exist. Mutations have been described in the RPE65, CRB1, RPGRIP1, AIPL1, GUCY2D, and CRX genes and other pedigrees show linkage to the LCA3 and LCA5 loci. The latter is a new locus which maps to 6q11-q16. The ocular findings and the evolution of the macula staphyloma are described in five members of a Pakistani family with consanguinity and a mutation in the LCA5 gene. METHODS: 13 family members including five affected individuals consented to DNA analysis and ocular examination including fundal photography. RESULTS: Ocular abnormalities are described. The most striking feature was the progression of macula abnormalities in three brothers resulting in a colobomatous appearance in the eldest compared to only mild atrophy in the youngest. The phenotypic pattern of this mutation in this Pakistani family contrasts with the "Old Order River Brethren" who were of Swiss descent, in whom the mutation was first described. CONCLUSION: The evolution of a new phenotypic picture is presented to a mutation in LCA5. PMID- 12642314 TI - Suppressive effects of selectin inhibitor SKK-60060 on the leucocyte infiltration during endotoxin induced uveitis. AB - BACKGROUND: It is well known that selectin is involved in the development of endotoxin induced uveitis (EIU), and has a major role in leucocyte infiltration. Recently, a novel selectin inhibitor (SKK-60060) that can block P and L selectins in vitro has been developed. This study was designed to investigate the anti inflammatory effects of SKK-60060 on the inflammatory reaction during EIU in rats by studying leucocyte-endothelium interactions. METHODS: EIU was induced in Lewis rats by footpad injection of lipopolysaccharide (LPS). SKK-60060 was administered 15 minutes before LPS injection, and its suppressive effects on inflammatory leucocyte behaviour were evaluated in vivo with acridine orange digital fluorography; the diameters of retinal arteries and veins were also measured. After these studies, aqueous humour was collected to evaluate leucocyte infiltration and protein leakage. RESULTS: After LPS injection, rolling leucocytes were observed in major retinal veins, followed by leucocyte infiltration into the vitreous cavity. Following treatment with SKK-60060, leucocyte rolling was significantly inhibited in the retinal veins (p <0.01), and subsequent leucocyte infiltration into the vitreous cavity was also significantly suppressed (p <0.01). Retinal vasodilation was also substantially suppressed in SKK-60060 treated rats (p <0.01). Similarly, leucocyte infiltration and protein leakage into the aqueous humour were reduced significantly by SKK-60060 (p <0.01). CONCLUSIONS: SKK-60060 treatment significantly inhibited the inflammatory reaction induced by LPS. Its inhibitory effects on P and L-selectin resulted in suppression of leucocyte infiltration and the subsequent inflammatory reaction caused by accumulated leucocytes. The current findings suggest that SKK-60060 may be useful in the management of uveitis. PMID- 12642315 TI - Lipopolysaccharide/interferon-gamma and not transforming growth factor beta inhibits retinal microglial migration from retinal explant. AB - BACKGROUND: /aims: The retina possesses a rich network of CD45(+) positive myeloid derived cells that both surround inner retinal vessels and lie within the retina (microglia). Microglia migrate and accumulate in response to neurodegeneration and inflammation. Although microglia express MHC class II, their role remains undefined. The aims of this study are to investigate changes in human microglia phenotype, migration, and activation status in response to pro inflammatory and anti-inflammatory stimulation. METHODS: Donor eyes were obtained from the Bristol Eye Bank with consent and whole retina was removed. 5 mm retinal trephines were cultured in glucose enhanced RPMI on cell culture insert membranes for up to 72 hours. The effects of lipopolysaccharide/interferon-gamma (LPS/IFNgamma) and transforming growth factor beta inhibits (TGFbeta) stimulation, alone or in combination, on migration, phenotype, and activation status (iNOS expression) of microglia were studied using immunofluorescence and cytokine analysis by ELISA. RESULTS: CD45(+) MHC class II(+) retinal microglia were observed within retinal explants, and in culture microglia readily migrated, adhered to culture membrane, downregulated MHC class II expression, and produced interleukin 12 (IL-12) and tumour necrosis factor alpha (TNFalpha). Following LPS/IFNgamma stimulation microglia remained MHC class II(-) iNOS(-), and secreted IL-10. Migration was suppressed and this could be reversed by neutralising IL-10 activity. TGFbeta did not affect ability of microglia to migrate and was unable to reverse LPS/IFNgamma induced suppression. CONCLUSIONS: Microglia readily migrate from retinal explants and are subsequently MHC class II(-), iNOS(-), and generate IL-12. In response to LPS/IFNgamma microglia produce IL-10, which inhibits both their migration and activation. TGFbeta was unable to counter LPS/IFNgamma effects. The data infer that microglia respond coordinately, dependent upon initial cytokine stimulation, but paradoxically respond to classic myeloid activation signals. PMID- 12642316 TI - Laser photocoagulation alters the pattern of staining for neurotrophin-4, GFAP, and CD68 in human retina. AB - AIMS: To investigate the staining pattern of neurotrophin-3 (NT3), neurotrophin-4 (NT4), and brain derived neurotrophic factor (BDNF) as well as glial fibrillary acid protein (GFAP) and CD68 in lasered human retina. METHODS: Retinal laser photocoagulation was performed on four patients (two males, two females) with choroidal malignant melanoma 1-6 days before enucleation. Three other enucleated eyes with malignant melanoma and three normal cadaveric donor eyes were used as controls. Immunohistochemistry was performed to investigate the pattern of staining of NT3, NT4, BDNF, GFAP, and CD68 in 7 mm sections of fixed specimens. RESULTS: Expression of NT4 was detected in the inner and outer nuclear layers of all the retinal sections examined but no NT3 and BDNF staining was seen. NT4 staining was found to be less intense in lasered and melanoma controls compared to normal cadaveric donor retinas. There was an upregulation of GFAP expression in both lasered and control eyes with melanoma in comparison with normal controls. CD68 staining was only observed in retinal pigment epithelium and choroid of lasered eyes. CONCLUSION: NT4 is expressed in inner and outer nuclear layers of normal human retina and its expression is downregulated following laser photocoagulation. This occurs in parallel with an increased expression of GFAP suggesting that reactive changes in Muller cells may be responsible for reduced NT4 staining. Expression of CD68 at the site of laser injury is consistent with a wound healing process as a response to local damage. PMID- 12642318 TI - Pigmentary retinopathy, macular oedema, and abnormal ERG with mitotane treatment. PMID- 12642319 TI - Wegener's granulomatosis. PMID- 12642320 TI - Coxsackievirus B4 associated uveoretinitis in an adult. PMID- 12642317 TI - Primary care and ophthalmology in the United Kingdom. AB - The National Health Service is now primary care led. There are different definitions for primary care and in this review they are analysed and related to ophthalmology to produce a working definition for ophthalmic primary care, summarised as the provision of first contact care for all ophthalmic conditions and follow up, preventive, and rehabilitative care of selected ophthalmic conditions, in a variety of settings, by a diverse workforce. The attributes of primary care are first contact, accessibility, continuity, longitudinality, comprehensiveness, coordination, equity, and accountability. The delivery of ophthalmic primary care should be governed by these and evaluated accordingly. The clinical content of primary care consists of the first presentation of disease, the management of minor illness and trauma, the recurrence of disease, the follow up and support of some chronic conditions, and the delivery of preventive health care. Planning for ophthalmic primary care needs to take service requirements of these categories of disease into account. Primary care research is abundant in ophthalmology but needs to be more structured and targeted. Ophthalmic primary care itself is urgently in need of recognition and formal adoption by the profession. PMID- 12642321 TI - Evaluation of telemedicine for slit lamp examination of the eye following cataract surgery. PMID- 12642322 TI - Inherited retinal dystrophy and asymmetric axial length. PMID- 12642323 TI - Adenoma of ciliary pigment epithelium: a case series. PMID- 12642324 TI - Phenylephrine 2.5% and 10% in phacoemulsification under topical anaesthesia: is there an effect on systemic blood pressure? PMID- 12642325 TI - Effect of preoperative detection of photoreceptor displacement on postoperative foveal findings in eyes with idiopathic macular hole. PMID- 12642326 TI - Nodular scleritis in a patient with sarcoidosis. PMID- 12642327 TI - Panophthalmitis following contact diode laser cyclophotocoagulation in a patient with failed trabeculectomy and trabeculotomy for congenital glaucoma. PMID- 12642328 TI - Bilaminar interepithelial bodies within fingerprint dystrophy-like changes in bilateral iridocorneal endothelial syndrome. PMID- 12642329 TI - Fine retinal crystalline deposits observed by confocal scanning laser ophthalmoscopic examination using infrared light. PMID- 12642330 TI - Tonic pupils from giant cell arteritis. PMID- 12642331 TI - Inflammatory optic neuropathy as the presenting feature of herpes simplex acute retinal necrosis. PMID- 12642332 TI - Frozen cucumber as a mount for processing vitreoretinal specimens. PMID- 12642333 TI - Cutaneous angiosarcoma of the eyelids. PMID- 12642334 TI - Those excrescences on Descemet's membrane. PMID- 12642335 TI - Permanent visual loss in a child with a rash. PMID- 12642336 TI - Acanthamoeba keratitis. PMID- 12642337 TI - A randomised controlled trial of written information. PMID- 12642338 TI - Surgery for glaucoma. PMID- 12642339 TI - Ocular and financial health. PMID- 12642341 TI - A (blood) smear campaign. PMID- 12642342 TI - Coincident expression of the chemokine receptors CCR6 and CCR7 by pathologic Langerhans cells in Langerhans cell histiocytosis. AB - It has been suggested that a switch in chemokine receptor expression underlies Langerhans cell migration from skin to lymphoid tissue. Activated cells are thought to down-regulate CCR6, whose ligand macrophage inflammatory protein-3 alpha (MIP-3 alpha)/CCL20 is expressed in skin, and up-regulate CCR7, whose ligands are in lymphoid tissues. In Langerhans cell histiocytosis (LCH), pathologic Langerhans cells (LCs) accumulate in several tissues, including skin, bone, and lymphoid organs. We have examined 24 LCH cases and find that pathologic LCs expressed CCR6 and CCR7 coincidentally in all cases. Furthermore, MIP-3 alpha/CCL20 is expressed by keratinocytes in involved skin and by macrophages and osteoblasts in involved bone. Expression of CCR6 by pathologic LCs may contribute to their accumulation in nonlymphoid organs such as skin and bone, whereas CCR7 expression may direct them to lymphoid tissue. Histiocytes in Rosai-Dorfman disease and hemophagocytic syndrome also coexpressed CCR6 and CCR7, suggesting that this may be a general attribute of abnormal histiocytes. PMID- 12642343 TI - Genetic and nutritional factors contributing to hyperhomocysteinemia in young adults. AB - A modestly elevated total plasma homocysteine concentration (tHcy) is generally accepted as an independent and graded risk factor for various pathologies, including vascular diseases, neural tube defects, Alzheimer disease, and pregnancy complications. We analyzed 5 common functional polymorphisms in enzymes involved in homocysteine metabolism (ie, methylenetetrahydrofolate reductase [MTHFR] 677C>T and 1298A>C, methionine synthase [MTR] 2756A>G, cystathionine beta synthase [CBS] 844ins68, and methionine synthase reductase [MTRR] 66A>G) in 452 young adults, and quantified their independent and interactive effects on tHcy concentrations. Serum folate, red cell folate, vitamin B(12), and tHcy concentrations were significantly influenced by MTHFR 677C>T genotypes. A particularly strong interaction was observed between the MTHFR 677TT genotype and serum folate, which led to a high tHcy phenotype that was more pronounced in males. The genetic contribution to the variance in tHcy was estimated to be approximately 9%, compared with approximately 35% that could be attributed to low folate and vitamin B(12). Our study indicates that dietary factors are centrally important in the control of tHcy levels in young adults with additional, but somewhat weaker, genetic effects. These data underscore the potential benefits that may be gained by improving the dietary status of young adults, and provide support for the implementation of folate/B-vitamin food fortification programs. PMID- 12642344 TI - Estimation of cell membrane alteration after drug treatment by LDH release. PMID- 12642345 TI - Diagnostic criteria for acute erythroleukemia. PMID- 12642346 TI - CYP2C9 exon 4 mutations and warfarin dose phenotype in Asians. PMID- 12642347 TI - The role of erythrocyte peroxiredoxin in detoxifying peroxides and in stimulating potassium efflux via the Gardos channels. PMID- 12642348 TI - Low levels of serum elastase are not associated with mutations in ELA-2 elastase encoding gene in chronic idiopathic neutropenia. PMID- 12642349 TI - Genetic clues to disease pathways in hypertrophic and dilated cardiomyopathies. PMID- 12642350 TI - Morning surge in blood pressure. PMID- 12642351 TI - NO balance: regulation of the cytoskeleton in congestive heart failure by nitric oxide. PMID- 12642352 TI - Glitazones and heart failure: critical appraisal for the clinician. PMID- 12642353 TI - Drugs that induce repolarization abnormalities cause bradycardia in zebrafish. AB - BACKGROUND: Drug-induced QT prolongation and torsades de pointes remain significant and often unpredictable clinical problems. Current in vitro preclinical assays are limited by biological simplicity, and in vivo models suffer from expense and low throughput. METHODS AND RESULTS: During a screen for the effects of 100 small molecules on the heart rate of the zebrafish, Danio rerio, we found that drugs that cause QT prolongation in humans consistently caused bradycardia and AV block in the zebrafish. Of 23 such drugs tested, 18 were positive in this initial screen. Poor absorption explained 4 of 5 false negative results, as demonstrated by microinjection. Overall, 22 of 23 compounds that cause repolarization abnormalities were positive in this assay. Antisense "knockdown" of the zebrafish KCNH2 ortholog yielded bradycardia in a dose dependent manner confirming the effects of reduction of repolarizing potassium current in this model. Classical drug-drug interactions between erythromycin and cisapride, as well as cimetidine and terfenadine, were also reproduced. CONCLUSION: This simple high-throughput assay is a promising addition to the repertoire of preclinical tests for drug-induced repolarization abnormalities. The genetic tractability of the zebrafish will allow the exploration of heritable modifiers of such drug effects. PMID- 12642354 TI - The VIVA trial: Vascular endothelial growth factor in Ischemia for Vascular Angiogenesis. AB - BACKGROUND: Recombinant human vascular endothelial growth factor protein (rhVEGF) stimulates angiogenesis in animal models and was well tolerated in Phase I clinical trials. VIVA (Vascular endothelial growth factor in Ischemia for Vascular Angiogenesis) is a double-blind, placebo-controlled trial designed to evaluate the safety and efficacy of intracoronary and intravenous infusions of rhVEGF. METHODS AND RESULTS: A total of 178 patients with stable exertional angina, unsuitable for standard revascularization, were randomized to receive placebo, low-dose rhVEGF (17 ng x kg(-1) x min(-1)), or high-dose rhVEGF (50 ng x kg(-1) x min(-1)) by intracoronary infusion on day 0, followed by intravenous infusions on days 3, 6, and 9. Exercise treadmill tests, angina class, and quality of life assessments were performed at baseline, day 60, and day 120. Myocardial perfusion imaging was performed at baseline and day 60. At day 60, the change in exercise treadmill test (ETT) time from baseline was not different between groups (placebo, +48 seconds; low dose, +30 seconds; high dose, +30 seconds). Angina class and quality of life were significantly improved within each group, with no difference between groups. By day 120, placebo-treated patients demonstrated reduced benefit in all three measures, with no significant difference compared with low-dose rhVEGF. In contrast, high-dose rhVEGF resulted in significant improvement in angina class (P=0.05) and nonsignificant trends in ETT time (P=0.15) and angina frequency (P=0.09) as compared with placebo. CONCLUSIONS: rhVEGF seems to be safe and well tolerated. rhVEGF offered no improvement beyond placebo in all measurements by day 60. By day 120, high-dose rhVEGF resulted in significant improvement in angina and favorable trends in ETT time and angina frequency. PMID- 12642355 TI - Cellular phospholipid and cholesterol efflux in high-density lipoprotein deficiency. AB - BACKGROUND: Prospective studies have examined the relationship between coronary artery disease and low plasma levels of high-density lipoprotein cholesterol (HDL C). METHODS AND RESULTS: We investigated the causes of hypoalphalipoproteinemia (HypoA; HDL-C <5th percentile) in 64 subjects (12 women and 52 men). Apolipoprotein AI-mediated cellular cholesterol and phospholipid efflux were measured in fibroblasts from HypoA subjects, 9 controls, 2 patients with Tangier disease, and 5 patients with hyperalphalipoproteinemia. A phospholipid efflux defect was defined as <70% of controls. Mean HDL-C was 0.49+/-0.21 mmol/L. Cholesterol and phospholipid efflux correlated strongly (r=0.72, P<0.001). Phospholipid efflux and HDL-C (r=0.64, P<0.001) correlated in HypoA subjects. However, phospholipid or cholesterol efflux was no longer a determinant of HDL-C levels at higher levels (> approximately 1.0 mmol/L) of HDL-C. In HypoA subjects, 4 cases of Tangier disease and 6 of familial HDL deficiency (heterozygous Tangier disease) were identified (10 of 64; 16%). In the remaining 54 subjects, mean lipid efflux was not significantly different from controls and subjects with hyperalphalipoproteinemia. A phospholipid efflux defect was identified in 7 additional HypoA subjects, and a cholesterol efflux defect was detected in 11 subjects. In 2 of these subjects, the ABCA1 gene was ruled out as the cause of the efflux defect, while in 3, the low HDL-C trait segregated with the ABCA1 gene locus. CONCLUSIONS: Lipidation of lipid-poor apolipoprotein AI may not be a major determinant of cholesterol accumulation within more mature HDL particles and increasing cholesterol or phospholipid efflux beyond normal levels may not lead to increase in plasma HDL-C levels. ABCA1 is essential in the initial steps of HDL formation but other plasma events are major modulators of HDL-C levels. PMID- 12642356 TI - Cardiac benefits of fish consumption may depend on the type of fish meal consumed: the Cardiovascular Health Study. AB - BACKGROUND: Few studies have examined associations of fish consumption with ischemic heart disease (IHD) risk among older adults or how different types of fish meals relate to IHD risk. METHODS AND RESULTS: In a population-based prospective cohort study, usual fish consumption was ascertained at baseline among 3910 adults aged > or =65 years and free of known cardiovascular disease in 1989 and 1990. Consumption of tuna and other broiled or baked fish correlated with plasma phospholipid long-chain n-3 fatty acids, whereas consumption of fried fish or fish sandwiches (fish burgers) did not. Over 9.3 years' mean follow-up, there were 247 IHD deaths (including 148 arrhythmic deaths) and 363 incident nonfatal myocardial infarctions (MIs). After adjustment for potential confounders, consumption of tuna or other broiled or baked fish was associated with lower risk of total IHD death (P for trend=0.001) and arrhythmic IHD death (P=0.001) but not nonfatal MI (P=0.44), with 49% lower risk of total IHD death and 58% lower risk of arrhythmic IHD death among persons consuming tuna/other fish 3 or more times per week compared with less than once per month. In similar analyses, fried fish/fish sandwich consumption was not associated with lower risk of total IHD death, arrhythmic IHD death, or nonfatal MI but rather with trends toward higher risk. CONCLUSIONS: Among adults aged > or =65 years, modest consumption of tuna or other broiled or baked fish, but not fried fish or fish sandwiches, is associated with lower risk of IHD death, especially arrhythmic IHD death. Cardiac benefits of fish consumption may vary depending on the type of fish meal consumed. PMID- 12642357 TI - Relation between platelet response to exercise and coronary angiographic findings in patients with effort angina. AB - BACKGROUND: Platelet reactivity is increased by exercise in patients with obstructive coronary artery disease (CAD) but not in patients with syndrome X. In this study, we prospectively investigated whether the platelet response to exercise might help distinguish, among patients with angina, those with obstructive CAD from those with normal coronary arteries (NCAs). METHODS AND RESULTS: Venous blood samples were collected before and 5 minutes after exercise from 194 consecutive patients with stable angina. Platelet reactivity was measured by the platelet function analyzer (PFA)-100 system as the time for flowing whole blood to occlude a collagen-adenosine diphosphate ring (closure time). Coronary angiography showed CAD in 163 patients (84%) and NCA in 31 patients (16%). Baseline closure time was shorter in NCA patients (78.0+/-16 versus 95.5+/-23 seconds, P<0.0001). With exercise, closure time decreased in CAD patients (-15.5 seconds; 95% confidence limits [CL], -13.0 to -18.0 seconds; P<0.0001), but increased in NCA patients (12.5 seconds; 95% CL, 7.4 to 17.7 seconds; P=0.0004). An increase in closure time with exercise > or =10 seconds had 100% specificity and positive predictive value for NCAs. Similarly, a decrease > or =10 seconds had 100% specificity and positive predictive value for CAD. A closure time change (increase or decrease) > or =10 seconds allowed a correct classification of 55% of all patients. CONCLUSIONS: Among patients with stable angina, the response of platelet reactivity to exercise was predictive of normal or stenosed coronary arteries at angiography. Specifically, an increase in closure time with exercise > or =10 seconds was invariably associated with the presence of NCA. PMID- 12642358 TI - Electron spin resonance characterization of vascular xanthine and NAD(P)H oxidase activity in patients with coronary artery disease: relation to endothelium dependent vasodilation. AB - BACKGROUND: Increased inactivation of nitric oxide by superoxide (O2*-) contributes to endothelial dysfunction in patients with coronary disease (CAD). We therefore characterized the vascular activities of xanthine oxidase and NAD(P)H oxidase, 2 major O2*--producing enzyme systems, and their relationship with flow-dependent, endothelium-mediated vasodilation (FDD) in patients with CAD. METHODS AND RESULTS: Xanthine- and NAD(P)H-mediated O*.- formation was determined in coronary arteries from 10 patients with CAD and 10 controls by using electron spin resonance spectroscopy. Furthermore, activity of endothelium bound xanthine oxidase in vivo and FDD of the radial artery were determined in 21 patients with CAD and 10 controls. FDD was measured before and after infusion of the antioxidant vitamin C (25 mg/min i.a.) to determine the portion of FDD inhibited by radicals. In coronary arteries from patients with CAD, xanthine- and NAD(P)H-mediated O2*- formation was increased compared with controls (xanthine: 12+/-2 versus 7+/-1 nmol O2*-/ microg protein; NADH: 11+/-1 versus 7+/-1 nmol O2* / microg protein; and NADPH: 12+/-2 versus 9+/-1 nmol O2*-/ microg protein; each P<0.05). Endothelium-bound xanthine oxidase activity was increased by >200% in patients with CAD (25+/-4 versus 9+/-1 nmol O2*-/ microL plasma per min; P<0.05) and correlated inversely with FDD (r=-0.55; P<0.05) and positively with the effect of vitamin C on FDD (r=0.54; P<0.05). CONCLUSIONS: The present study represents the first electron spin resonance measurements of xanthine and NAD(P)H oxidase activity in human coronary arteries and supports the concept that increased activities of both enzymes contribute to increased vascular oxidant stress in patients with CAD. Furthermore, the present study suggests that increased xanthine oxidase activity contributes to endothelial dysfunction in patients with CAD and may thereby promote the atherosclerotic process. PMID- 12642359 TI - Mutations in the human muscle LIM protein gene in families with hypertrophic cardiomyopathy. AB - BACKGROUND: Muscle LIM protein (MLP) is an essential nuclear regulator of myogenic differentiation. Additionally, it may act as an integrator of protein assembly of the actin-based cytoskeleton. MLP-knockout mice develop a marked cardiac hypertrophy reaction and dilated cardiomyopathy (DCM). MLP is therefore a candidate gene for heritable forms of hypertrophic cardiomyopathy (HCM) and DCM in humans. METHODS AND RESULTS: We analyzed 1100 unrelated individuals (400 patients with DCM, 200 patients with HCM, and 500 controls) for mutations in the human CRP3 gene that encodes MLP. We found 3 different missense mutations in 3 unrelated patients with familial HCM but detected no mutation in the DCM group or the controls. All mutations predicted an amino acid exchange at highly conserved residues in the functionally important LIM1 domain, which is responsible for interaction with alpha-actinin and with certain muscle-specific transcription factors. Protein-binding studies indicate that mutations in the CRP3 gene lead to a decreased binding activity of MLP to alpha-actinin. All 3 index patients were characterized by typical asymmetrical septal hypertrophy. Family studies revealed cosegregation of clinically affected individuals with the respective mutations in MLP. CONCLUSION: Here, we present evidence that mutations in the CRP3/MLP gene can cause HCM. PMID- 12642360 TI - Raised sympathetic nerve activity in heart failure and central sleep apnea is due to heart failure severity. AB - BACKGROUND: Congestive heart failure (CHF) patients with central sleep apnea (CHF CSA) have elevated plasma norepinephrine (NE) compared with CHF patients without apnea (CHF-N). Patients with CHF-CSA also demonstrate higher mean pulmonary artery pressure (PAP), which is suggestive of worse cardiac function. Whether CSA contributes to chronic elevation of sympathetic nerve activity or is associated with more severe CHF remains unknown. We measured awake total body and cardiac NE spillover and related these to measurements of cardiac hemodynamics and apnea severity in CHF patients with CSA, with normal breathing, and with obstructive sleep apnea (CHF-OSA). METHODS AND RESULTS: A total of 55 CHF patients underwent right heart catheterization and measurements of total body and cardiac NE spillover using NE radioisotope dilution methodology. After polysomnography, patients were grouped by apnea type: 19 were CHF-N, 15 were CHF-OSA, and 21 were CHF-CSA. Compared with the CHF-N and CHF-OSA groups, the CHF-CSA group had significantly higher total body NE spillover (4.62+/-0.56 versus 4.47+/-0.54 versus 6.95+/-0.89 nmol/min, respectively; P=0.03), cardiac NE spillover (0.25+/ 0.05 versus 0.21+/-0.05 versus 0.42+/-0.06 nmol/min, respectively; P=0.02) and mean PAP (23.5+/-2.4 versus 21.2+/-0.8 versus 30.4+/-0.2 mm Hg, respectively; P<0.02). However, controlling for severity of CHF resulted in no significant differences in NE kinetics among the 3 groups. In a stepwise regression, only mean PAP independently correlated with total body (r=0.33, P=0.03) and cardiac NE spillover (r=0.44, P=0.002). Sleep apnea severity bore no relationship to markers of sympathetic nerve activity. CONCLUSION: Total body and cardiac sympathetic nerve activity are elevated in CHF-CSA compared with CHF-OSA and CHF-N patients and are related to heart failure not apnea severity. PMID- 12642361 TI - Morning surge in blood pressure as a predictor of silent and clinical cerebrovascular disease in elderly hypertensives: a prospective study. AB - BACKGROUND: Cardiovascular events occur most frequently in the morning hours. We prospectively studied the association between the morning blood pressure (BP) surge and stroke in elderly hypertensives. METHODS AND RESULTS: We studied stroke prognosis in 519 older hypertensives in whom ambulatory BP monitoring was performed and silent cerebral infarct was assessed by brain MRI and who were followed up prospectively. The morning BP surge (MS) was calculated as follows: mean systolic BP during the 2 hours after awakening minus mean systolic BP during the 1 hour that included the lowest sleep BP. During an average duration of 41 months (range 1 to 68 months), 44 stroke events occurred. When the patients were divided into 2 groups according to MS, those in the top decile (MS group; MS > or =55 mm Hg, n=53) had a higher baseline prevalence of multiple infarcts (57% versus 33%, P=0.001) and a higher stroke incidence (19% versus 7.3%, P=0.004) during the follow-up period than the others (non-MS group; MS <55 mm Hg, n=466). After they were matched for age and 24-hour BP, the relative risk of the MS group versus the non-MS group remained significant (relative risk=2.7, P=0.04). The MS was associated with stroke events independently of 24-hour BP, nocturnal BP dipping status, and baseline prevalence of silent infarct (P=0.008). CONCLUSIONS: In older hypertensives, a higher morning BP surge is associated with stroke risk independently of ambulatory BP, nocturnal BP falls, and silent infarct. Reduction of the MS could thus be a new therapeutic target for preventing target organ damage and subsequent cardiovascular events in hypertensive patients. PMID- 12642362 TI - Cyclooxygenase-2 expression and role of vasoconstrictor prostanoids in small mesenteric arteries from patients with Crohn's disease. AB - BACKGROUND: The present study investigates the vascular reactivity and the involvement of nitric oxide and prostanoids in regulating vasoconstriction of small mesenteric arteries from patients with Crohn's disease (CD) to understand the vascular component of this pathology. METHODS AND RESULTS: An increased production of proinflammatory cytokines (tumor necrosis factor-alpha and interleukins 1beta, 6, and 8) has been observed in biopsy specimens of inflammatory intestinal mucosa. However, contractile responses of small mesenteric arteries from CD patients in response to norepinephrine were not changed ex vivo when compared with controls. Exposure to either the nitric oxide synthase inhibitor N(G)-nitro-L-arginine or the cyclooxygenase (COX) inhibitor indomethacin did not modify contractions induced by norepinephrine in either control or CD patients. However, in the latter, the specific COX-2 inhibitor N-(2 cyclohexyloxy-4-nitrophenyl) methanesulfonamide significantly attenuated norepinephrine-induced vasoconstriction. Furthermore, immunohistochemical analysis showed marked COX-2 expression in the whole arterial wall of vessels from CD patients. Vessels from control patients exhibited weak COX-2 staining in the adventitial and endothelial layers only. CONCLUSIONS: The above results provide direct evidence for COX-2 expression in small mesenteric arteries from CD patients. They also shed new light on the involvement of vasoconstrictor metabolites of COX in regulating contraction of these arteries. Of particular interest is the balance between vasoconstrictor products from COX-2 and unidentified vasodilatory products that maintained vascular reactivity in a physiological range despite an increase of circulatory cytokines in patients with CD. PMID- 12642363 TI - Angiogenic property of hepatocyte growth factor is dependent on upregulation of essential transcription factor for angiogenesis, ets-1. AB - BACKGROUND: Although hepatocyte growth factor (HGF) is an angiogenic growth factor, it is still unclear how it exerts its angiogenic effects. Thus, we focused on the role of an essential transcription factor for angiogenesis, ets-1. In this study, we addressed the following specific questions: (1) what genes responsible for angiogenesis can be regulated by HGF and (2) whether upregulation of gene expression for angiogenesis is dependent on ets-1. METHODS AND RESULTS: In human endothelial cells, HGF significantly stimulated the matrix-degrading pathway, such as the production of matrix metalloprotease-1 (MMP-1) through its specific receptor, c-met. In addition, HGF also significantly increased HGF itself and its specific receptor, c-met. Moreover, HGF significantly increased the transcription activity and mRNA expression of ets-1 in a time-dependent manner. Importantly, transfection of antisense ets-1 oligodeoxynucleotides (ODN) resulted in a significant reduction in MMP-1, HGF and c-met. Interestingly, HGF also stimulated ets-1 mRNA in vascular smooth muscle cells, similar to endothelial cells. Of importance, transfection of antisense ets-1 ODN resulted in a significant decrease in vascular endothelial growth factor (VEGF) and HGF expression, whereas HGF stimulated both HGF and VEGF expression. Moreover, in vivo transfection of ets-1 antisense ODN resulted in an inhibition of angiogenesis induced by the HGF gene in a rat ischemic hindlimb model. CONCLUSIONS: Here, we demonstrated that HGF stimulated the expression of MMP-1, VEGF, HGF itself, and c-met in human endothelial cells and vascular smooth muscle cells. Upregulation of angiogenesis-related genes was largely dependent on the induction of ets, especially ets-1. These data provide new information about the mechanisms of angiogenesis. PMID- 12642364 TI - Oxidative stress mediates tumor necrosis factor-alpha-induced mitochondrial DNA damage and dysfunction in cardiac myocytes. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) and angiotensin II (Ang II) are implicated in the development and further progression of heart failure, which might be, at least in part, mediated by the production of reactive oxygen species (ROS). However, the cause and consequences of this agonist-mediated ROS production in cardiac myocytes have not been well defined. Recently, we demonstrated that increased ROS production was associated with mitochondrial DNA (mtDNA) damage and dysfunction in failing hearts. We thus investigated whether the direct exposure of cardiac myocytes to TNF-alpha and Ang II in vitro could induce mtDNA damage via production of ROS. METHODS AND RESULTS: TNF-alpha increased ROS production within cultured neonatal rat ventricular myocytes after 1 hour, as assessed by 2',7'-dichlorofluorescin diacetate fluorescence microscopy. TNF-alpha also decreased mtDNA copy number by Southern blot analysis in association with complex III activity, which was prevented in the presence of the antioxidant alpha-tocopherol. A direct exposure of myocytes to H2O2 caused a similar decrease in mtDNA copy number. In contrast, Ang II did not affect mtDNA copy number, despite the similar increase in ROS production. TNF-alpha-mediated ROS production and a decrease in mtDNA copy number were inhibited by the sphingomyelinase inhibitor D609. Furthermore, N-acetylsphingosine (C2-ceramide), a synthetic cell-permeable ceramide analogue, increased myocyte ROS production, suggesting that TNF-alpha-mediated ROS production and subsequent mtDNA damage were mediated by the sphingomyelin-ceramide signaling pathway. CONCLUSIONS: The intimate link between TNF-alpha, ROS, and mtDNA damage might play an important role in myocardial remodeling and failure. PMID- 12642365 TI - Downregulation of cytoskeletal muscle LIM protein by nitric oxide: impact on cardiac myocyte hypertrophy. AB - BACKGROUND: In chronic heart failure, myocardial expression of the inducible isoform of nitric oxide (NO) synthase (NOS2) is enhanced, leading to a sustained production of NO. We postulated that NO modulates expression of genes in cardiac myocytes that may be functionally important in the context of cardiac hypertrophy and failure. METHODS AND RESULTS: As revealed by cDNA expression array analyses, the NO donor SNAP, which has been shown previously to inhibit agonist-induced cardiac myocyte hypertrophy, downregulates expression of the cytoskeleton associated muscle LIM protein (MLP) in endothelin-1 (ET-1)-stimulated neonatal rat cardiac myocytes. Northern blotting and immunoblotting experiments confirmed this finding and established that SNAP negatively controls MLP mRNA (-49%, P<0.01) and protein (-52%, P<0.01) abundance in ET-1-treated cardiomyocytes via cGMP-dependent protein kinase and superoxide/peroxynitrite-dependent signaling pathways. Treatment of cardiac myocytes with IL-1beta and IFN-gamma downregulated MLP expression levels via induction of NOS2. Moreover, expression levels of NOS2 and MLP were inversely correlated in the failing human heart, indicating that NOS2 may regulate MLP abundance in vitro and in vivo. Antisense oligonucleotides were used to explore the functional consequences of reduced MLP expression levels in cardiac myocytes. Like SNAP, antisense downregulation of MLP protein expression (-52%, P<0.01) blunted the increases in protein synthesis, cell size, and sarcomere organization in response to ET-1 stimulation. Conversely, overexpression of MLP augmented cell size and sarcomere organization in cardiac myocytes. CONCLUSIONS: NO negatively controls MLP expression in cardiac myocytes. Because MLP is necessary and sufficient for hypertrophy and sarcomere assembly, MLP downregulation may restrain hypertrophic growth in pathophysiological situations with increased cardiac NO production. PMID- 12642366 TI - Electrical and contractile remodeling during the first days of atrial fibrillation go hand in hand. AB - BACKGROUND: The mechanisms of the atrial contractile dysfunction induced by atrial fibrillation (AF) are not completely understood. In particular, the relation between the atrial dysfunction and electrical remodeling has not yet been studied. METHODS AND RESULTS: Seven goats were chronically instrumented with electrodes sutured to the atria and with ultrasonic piezoelectric crystals to record the atrial diameters. A pressure transducer was implanted in the right atrium. After 5 minutes, 3 hours, and throughout the first 5 days of artificially maintained AF, atrial contractile function was measured and the atrial effective refractory period (AERP) was monitored for comparison. Also, the positive inotropic effects of the L-type Ca2+-channel agonist BayY5959 and short trains of rapid atrial pacing were studied. After resumption of sinus rhythm, the recovery of atrial contractile function was followed. After 5 minutes of AF, atrial contractility was decreased by approximately 55% but recovered completely within 10 minutes. Five days of AF nearly completely abolished the atrial contractile function, and recovery took 2 days. During the first days of AF, the development of the contractile dysfunction followed the same time course as the shortening of AERP (electrical remodeling). In remodeled atria, BayY5959 increased atrial contractility to the same extent as it prolonged AERP. The inotropic effect of short trains of rapid atrial pacing was similar in normal and remodeled atria. CONCLUSIONS: Depending on the duration of AF, different mechanisms contribute to the AF-induced atrial hypocontractility. Atrial contractile remodeling during several days of AF goes hand in hand with electrical remodeling and might be caused by a reduction of the L-type Ca2+-current. PMID- 12642367 TI - Effects of antiarrhythmic drugs on fibrillation in the remodeled atrium: insights into the mechanism of the superior efficacy of amiodarone. AB - BACKGROUND: The basis of the unique effectiveness of amiodarone for atrial fibrillation (AF) is poorly understood. The present study tested the hypothesis that amiodarone blocks electrical remodeling induced by atrial tachycardia. METHODS AND RESULTS: Mongrel dogs were subjected to atrial tachycardia (400 bpm for 7 days) in the absence and presence of therapy with amiodarone, the class III cardiac antiarrhythmic drug dofetilide, or the class I agent flecainide begun 3 days before the onset of tachypacing and maintained until a final electrophysiological study. AF vulnerability (percentage of sites with AF induction by single premature extrastimuli), mean AF duration, atrial effective refractory period (ERP), and conduction velocity were compared among these dogs and in unpaced dogs in the absence or presence of treatment with the same agents. Only amiodarone prevented promotion of AF duration and vulnerability by atrial tachycardia. Furthermore, only amiodarone eliminated tachycardia-induced ERP abbreviation and loss of ERP rate adaptation while obviating L-type Ca2+-current alpha1c-subunit downregulation as determined by Western blot. In an additional series of dogs monitored with repeated electrophysiological studies, amiodarone administered after the induction of atrial tachycardia remodeling reversed remodeling within several days, despite continued atrial tachypacing during amiodarone therapy. CONCLUSIONS: Amiodarone is uniquely effective against AF promotion by atrial tachycardia remodeling in this experimental model and prevents electrophysiological and biochemical consequences of remodeling. Amiodarone also reversed remodeling established by 4 days of atrial tachycardia. The inhibition of atrial tachycardia remodeling may therefore contribute to the superior efficacy of amiodarone in AF. PMID- 12642368 TI - Images in cardiovascular medicine. Late internal mammary arteriovenous fistula. PMID- 12642369 TI - Obesity, insulin resistance, diabetes, and cardiovascular risk in children: an American Heart Association scientific statement from the Atherosclerosis, Hypertension, and Obesity in the Young Committee (Council on Cardiovascular Disease in the Young) and the Diabetes Committee (Council on Nutrition, Physical Activity, and Metabolism). PMID- 12642370 TI - Images in cardiovascular medicine. Surgical left ventricular restoration: an extreme case. PMID- 12642371 TI - Beware that morning surge. PMID- 12642372 TI - Commentary on the article presented by Murdoch et al. PMID- 12642373 TI - The vasodilatory action of testosterone: a potassium-channel opening or a calcium antagonistic action? PMID- 12642374 TI - Preservation of vascular contraction during ageing: dual effect on calcium handling and sensitization. AB - (1) The present study was aimed to characterize the effects of ageing on vascular contraction by noradrenaline in rat isolated arteries. The existence of vascular bed heterogeneity was investigated in endothelium-denuded conductance (aorta) and resistance (small mesenteric artery, SMA) arteries, with respect to Ca(2+) handling, Ca(2+) sensitization or Ca(2+)-independent mechanisms. (2) In both arteries, contractions to noradrenaline were not different between adult and aged rats. (3) In Ca(2+)- free medium, noradrenaline elicited a transient increase in tension that was reduced by the Ca(2+) mobilizing agents, ryanodine and thapsigargin, in arteries from adult rats. A loss of the thapsigargin- but not the ryanodine-sensitive component of noradrenaline-induced contraction was observed in the two arteries from aged rats. (4) After depletion of Ca(2+) stores with noradrenaline, addition of exogenous CaCl(2) produced a sustained contraction that was decreased to the same extent by the protein kinase C inhibitor, GF 109203X and the tyrosine kinase inhibitor, tyrphostin A-23, in arteries from adult and aged rats. The Rho-associated protein kinase inhibitor, Y 27632, caused identical relaxation of noradrenaline pre-contracted arteries from both age groups. (5) Basal intracellular calcium ([Ca(2+)](i)) was higher in SMA from aged than from adult rats. In addition, the noradrenaline [Ca(2+)](i)-force relationship was significantly shifted to the right in the SMA from aged rats. (6) Altogether, these data indicate that responsiveness to noradrenaline is preserved both in conductance and resistance arteries with ageing. The latter results from the association of increased basal [Ca(2+)](i), changes in Ca(2+) handling at the level of thapsigargin-sensitive sarcoplasmic reticulum Ca(2+) ATPases and decreased myofilament sensitivity to Ca(2+). PMID- 12642375 TI - Identification of MEK1 as a novel target for the treatment of neuropathic pain. AB - (1) In the present study we have attempted to identify changes in gene expression which are associated with neuropathic pain using subtractive suppression hybridization analysis of the lumbar spinal cord of animals suffering streptozocin induced diabetic neuropathy. (2) Using this approach, we found a significant up-regulation of several key components of the extracellular signal regulated kinase (ERK) cascade. These findings were confirmed by Western blot analysis, which demonstrated that the levels of active ERK1 and 2 correlated with the onset of streptozocin-induced hyperalgesia. (3) Intrathecal administration of the selective MAPK/ERK-kinase (MEK) inhibitor PD 198306 dose-dependently (1-30 micro g) blocked static allodynia in both the streptozocin and the chronic constriction injury (CCI) models of neuropathic pain. (4) The antihyperalgesic effects of PD 198306, in both the streptozocin and CCI models of neuropathic pain, correlated with a reduction in the elevated levels of active ERK1 and 2 in lumbar spinal cord. (5) Intraplantar administration of PD 198306 had no effect in either model of hyperalgesia, indicating that changes in the activation of ERKs and the effect of MEK inhibition are localized to the central nervous system. (6) In summary, we have demonstrated for the first time that the development of neuropathic pain is associated with an increase in the activity of the MAPK/ERK kinase cascade within the spinal cord and that enzymes in this pathway represent potential targets for the treatment of this condition. PMID- 12642376 TI - Expression and functional role of Rho-kinase in rat urinary bladder smooth muscle. AB - (1) The involvement of Rho-kinase (ROCK) in the contractile mechanisms mediating smooth muscle contraction of the rat urinary bladder was investigated using expression studies and the ROCK inhibitor Y-27632. (2) Both isoforms of ROCK (ROCK I and ROCK II) were detected in high levels in rat urinary bladder. (3) Y 27632 (10 micro M) significantly attenuated contractions of rat urinary bladder strips evoked by the G-protein coupled receptor agonists carbachol (58.1+/-10.5% at 0.3 micro M) and neurokinin A (68.6+/-12.7% at 1 micro M) without affecting contractions to potassium chloride (10-100 mM). In addition, basal tone was reduced by 47.8+/-2.0% by 10 micro M Y-27632 in the absence of stimulation. (4) Contractions of urinary bladder strips evoked by the P2X receptor agonist alpha,beta-methylene ATP (alpha,beta-mATP; 10 micro M) were also attenuated by Y 27632 (30.0+/-7.2% at 10 micro M). (5) Y-27632 (10 micro M) significantly attenuated contractions evoked by electrical field stimulation (2-16 Hz). The effect of Y-27632 on the tonic portion of the neurogenic response (4-16 Hz) was not significantly different from the effect of atropine (1 micro M) alone. (6) While the mechanism underlying the ability of Y-27632 to inhibit alpha,beta-mATP evoked contractions remains undetermined, the results of the present study clearly demonstrate a role for ROCK in the regulation of rat urinary bladder smooth muscle contraction and tone. PMID- 12642377 TI - Cannabinoid receptor-independent inhibition by cannabinoid agonists of the peripheral 5-HT3 receptor-mediated von Bezold-Jarisch reflex. AB - (1) On the basis of previous findings that cannabinoids inhibit the function of human and rat 5-HT(3) receptors in vitro, we investigated whether cannabinoid receptor agonists also modulate the activity of the rat peripheral 5-HT(3) receptors on the terminals of cardiopulmonary afferent C-fibres in vivo. (2) In urethane-anaesthetized rats, pre-treated intravenously (i.v.) with the CB(1) receptor antagonist SR 141716A (3 micro mol kg(-1)) and with the beta(1)/beta(2) adrenoceptor antagonist propranolol (0.3-0.4 micro mol kg(-1)), bolus injection of the serotonin 5-HT(3) receptor agonist phenylbiguanide (3-10 micro g kg(-1), i.v.) or the vanilloid VR1 receptor agonist capsaicin (3-10 micro g kg(-1), i.v.) caused an immediate decrease in heart rate and mean arterial blood pressure (the von Bezold-Jarisch reflex). (3) The phenylbiguanide-induced bradycardia was dose dependently attenuated by the cannabinoid receptor agonists CP 55,940 (0.1-1 micro mol kg(-1), i.v.) and WIN 55,212-2 (0.1-3 micro mol kg(-1), i.v.) 20 min after injection, but not by the inactive S-(-)enantiomer of the latter, WIN 55,212-3 (1 micro mol kg(-1), i.v.). The inhibition was reversible within 30 min. The extent of inhibition by the highest doses of cannabinoid receptor agonists amounted to about 50%. Both cannabinoid receptor agonists failed to affect the capsaicin-evoked bradycardia. (4) In conclusion, our results demonstrate that cannabinoid receptor agonists modulate the von Bezold-Jarisch reflex by inhibiting peripheral serotonin 5-HT(3) receptors in rats in vivo. An analogous mechanism of cannabinoid receptor agonists may be assumed to be involved in other serotonin 5-HT(3) receptor-mediated responses. PMID- 12642378 TI - Functional coupling of the human dopamine D2 receptor with G alpha i1, G alpha i2, G alpha i3 and G alpha o G proteins: evidence for agonist regulation of G protein selectivity. AB - (1) The human dopamine D(2long) (D(2L)) receptor was expressed with four different G proteins in Sf9 cells using the baculovirus expression system. When co-expressed with G(i)/G(o) G proteins (G(i1)alpha, G(i2)alpha, G(i3)alpha, or G(o)alpha, plus Gbeta(1) and Ggamma(2)), the receptor displayed a high-affinity binding site for the agonists (dopamine and NPA), which was sensitive to GTP (100 micro M), demonstrating interaction between the receptor and the different G proteins. (2) The receptor to G protein ratio (R : G ratio) was evaluated using [(3)H]-spiperone saturation binding (R) and [(35)S]-GTPgammaS saturation binding (G). R : G ratios of 1 : 12, 1 : 3, 1 : 14 and 1 : 5 were found for G(i1), G(i2), G(i3), and G(o) preparations, respectively. However, when R : G ratios of 1 : 2 and 1 : 12 were compared for G(i2) and G(o), no difference was found for the stimulation of [(35)S]-GTPgammaS binding. (3) Several agonists were tested for their ability to stimulate [(35)S]-GTPgammaS binding to membranes co-expressing the receptor and various G proteins. All the compounds tested showed agonist activity in preparations expressing G(i3) and G(o). However, for G(i2) and G(i1) preparations, compounds such as S-(-)-3-PPP and p-tyramine were unable to stimulate [(35)S]-GTPgammaS binding. (4) Most of the compounds showed higher relative efficacies (compared to dopamine) and higher potencies in the preparation expressing G(o). Comparison of the effects of different agonists in the different preparations showed that each agonist differentially activates the four G proteins. (5) We conclude that the degree of selectivity of G protein activation by the D(2L) receptor can depend on the conformation of the receptor stabilised by an agonist. PMID- 12642379 TI - Different domains of the glucagon and glucagon-like peptide-1 receptors provide the critical determinants of ligand selectivity. AB - (1) Glucagon and glucagon-like peptide-1 (GLP-1) are homologous peptide hormones with important functions in glucose metabolism. The receptors for glucagon and GLP-1 are homologous family B G-protein coupled receptors. The GLP-1 receptor amino-terminal extracellular domain is a major determinant of glucagon/GLP-1 selectivity of the GLP-1 receptor. However, the divergent residues in glucagon and GLP-1 that determine specificity for the GLP-1 receptor amino-terminal extracellular domain are not known. Less is known about how the glucagon receptor distinguishes between glucagon and GLP-1. (2) We analysed chimeric glucagon/GLP-1 peptides for their ability to bind and activate the glucagon receptor, the GLP-1 receptor and chimeric glucagon/GLP-1 receptors. The chimeric peptide GLP-1(7 20)/glucagon(15-29) was unable to bind and activate the glucagon receptor. Substituting the glucagon receptor core domain with the GLP-1 receptor core domain (chimera A) completely rescued the affinity and potency of GLP-1(7 20)/glucagon(15-29) without compromising the affinity and potency of glucagon. Substituting transmembrane segment 1 (TM1), TM6, TM7, the third extracellular loop and the intracellular carboxy-terminus of chimera A with the corresponding glucagon receptor segments re-established the ability to distinguish GLP-1(7 20)/glucagon(15-29) from glucagon. Corroborant results were obtained with the opposite chimeric peptide glucagon(1-14)/GLP-1(21-37). (3) The results suggest that the glucagon and GLP-1 receptor amino-terminal extracellular domains determine specificity for the divergent residues in the glucagon and GLP-1 carboxy-terminals respectively. The GLP-1 receptor core domain is not a critical determinant of glucagon/GLP-1 selectivity. Conversely, the glucagon receptor core domain contains two or more sub-segments which strongly determine specificity for divergent residues in the glucagon amino-terminus. PMID- 12642380 TI - 5-HT moduline: an endogenous inhibitor of 5-HT(1B/1D)-mediated contraction in pulmonary arteries. AB - (1) 5-HT moduline (5-HTm) is tetrapeptide (Leu-Ser-Ala-Leu) previously shown to act as a specific endogenous antagonist to central 5-HT(1B/1D) receptors. Its effects were investigated in rat and rabbit pulmonary arteries (PAs). (2) In rabbit PAs, contractile responses to the 5-HT(1B/1D) receptor agonist 5 carboxamidotryptamine (5-CT) were inhibited by 1 and 10 micro M 5-HTm in a non competitive fashion with the maximum contractile response (E(max), per cent of response to 50 mM KCl) being reduced from 65.6+/-7% (n=6) to 39.7+/-6.5% (n=6) and 25.2+/-7.9 (n=4), respectively. The ability of 5-HTm to inhibit responses to 5-CT was increased by the aminopeptidase inhibitor bestatin (10 micro M). (3) In the rabbit PAs, the nitric oxide synthase inhibitor, N(omega)-nitro-L-arginine methylester (L-NAME) potentiated responses to 5-CT (E(max): 106+/-22.5 (n=4)) and this response was also inhibited by 10 micro M 5-HTm (E(max): 38+/-13% (n=8)). (4) 5-HTm (10 micro M) inhibited responses to 5-CT in rat PAs, the E(max) being reduced from 24.8+/-4.1% (n=7) to 15.5+/-3.7% (n=9). 5-HTm induced relaxation of 5-CT-pre-constricted rat PAs with a pIC(50) of 9.0+/-0.6 (n=9). (5) In PAs from chronic hypoxic, pulmonary hypertensive rats, the maximum response to 5-CT was increased to 80+/-8.5% (n=11). 5-HTm reduced this response to 34.4+/-6.3% (n=12). L-NAME markedly inhibited the ability of 5-HTm to inhibit responses to 5-CT (E(max) before 5-HTm: 100.5+/-16% (n=5), E(max) after 5-HTm: 107+/-11.3% (n=4)). (6) In conclusion we show here for the first time that 5-HTm is a non-competitive inhibitor of 5-HT(1B/1D) receptor-mediated constriction in PAs. In rat PAs, L NAME can inhibit this effect of 5-HTm. PMID- 12642381 TI - Interactions of atropine with heterologously expressed and native alpha 3 subunit containing nicotinic acetylcholine receptors. AB - (1) Atropine, a classical muscarinic antagonist, has been reported previously to inhibit neuronal nicotinic acetylcholine receptors (nAChRs). In the present study, the action of atropine has been examined on alpha3beta4 receptors expressed heterologously in Xenopus oocytes and native nAChRs in medial habenula neurons. (2) At concentrations of atropine often used to inhibit muscarinic receptors (1 micro M), responses induced by near-maximal nicotine concentrations (100 micro M) at negative holding potentials (-65 mV) are inhibited (14-30%) in a reversible manner in both alpha4 and alpha3 subunit-containing heteromeric nAChRs. Half-maximal effective concentrations (IC(50) values) for atropine inhibition are similar for the four classes of heteromeric receptors studied (4 13 micro M). (3) For alpha3beta4 nAChRs in oocytes, inhibition by atropine (10 micro M) is not overcome at higher concentrations of agonist, and is increased with membrane hyperpolarization. These results are consistent with non competitive antagonism--possibly ion channel block. (4) At low concentrations of both nicotine (10 micro M) and atropine (<10 micro M), potentiation ( approximately 25%) of alpha3beta4 nAChR responses in oocytes is observed. The relative balance between potentiation and inhibition is dependent upon membrane potential. (5) In rat medial habenula (MHb) neurons, atropine (0.3-3.0 micro M) inhibited nicotine-induced responses in both a concentration and membrane potential-dependent manner (at -40 mV, IC(50)=4 micro M), similar to the effects on alpha3beta4-nAChRs in oocytes. However, unlike heterologously expressed receptors, potentiation was barely detectable at depolarized membrane potentials using low concentrations of nicotine (3-10 micro M). Conversely, the weak agonist, choline (1-3 mM) was observed to augment responses of MHb nAChRs. PMID- 12642382 TI - Ajoene, a garlic compound, inhibits protein prenylation and arterial smooth muscle cell proliferation. AB - (1) Ajoene is a garlic compound with anti-platelet properties and, in addition, was shown to inhibit cholesterol biosynthesis by affecting 3-hydroxy-3-methyl glutaryl coenzyme A (HMG-CoA) reductase and late enzymatic steps of the mevalonate (MVA) pathway. (2) MVA constitutes the precursor not only of cholesterol, but also of a number of non-sterol isoprenoids, such as farnesyl and geranylgeranyl groups. Covalent attachment of these MVA-derived isoprenoid groups (prenylation) is a required function of several proteins that regulate cell proliferation. We investigated the effect of ajoene on rat aortic smooth muscle cell proliferation as related to protein prenylation. (3) Cell counting, DNA synthesis, and cell cycle analysis showed that ajoene (1-50 micro M) interfered with the progression of the G1 phase of the cell cycle, and inhibited rat SMC proliferation. (4) Similar to the HMG-CoA reductase inhibitor simvastatin, ajoene inhibited cholesterol biosynthesis. However, in contrast to simvastatin, the antiproliferative effect of ajoene was not prevented by the addition of MVA, farnesol (FOH), and geranylgeraniol (GGOH). Labelling of smooth muscle cell cellular proteins with [3H]-FOH and [3H]-GGOH was significantly inhibited by ajoene. (5) In vitro assays for protein farnesyltransferase (PFTase) and protein geranylgeranyltransferase type I (PGGTase-I) confirmed that ajoene inhibits protein prenylation. High performance liquid chromatography (HPLC) and mass spectrometry analyses also demonstrated that ajoene causes a covalent modification of the cysteine SH group of a peptide substrate for protein PGGTase I. (6) Altogether, our results provide evidence that ajoene interferes with the protein prenylation reaction, an effect that may contribute to its inhibition of SMC proliferation. PMID- 12642383 TI - Identification of the di-pyridyl ketone isonicotinoyl hydrazone (PKIH) analogues as potent iron chelators and anti-tumour agents. AB - (1) In an attempt to develop chelators as potent anti-tumour agents, we synthesized two series of novel ligands based on the very active 2 pyridylcarboxaldehyde isonicotinoyl hydrazone (PCIH) group. Since lipophilicity and membrane permeability play a critical role in Fe chelation efficacy, the aldehyde moiety of the PCIH series, namely 2-pyridylcarboxaldehyde, was replaced with the more lipophilic 2-quinolinecarboxaldehyde or di-2-pyridylketone moieties. These compounds were then systematically condensed with the same group of acid hydrazides to yield ligands based on 2-quinolinecarboxaldehyde isonicotinoyl hydrazone (QCIH) and di-2-pyridylketone isonicotinoyl hydrazone (PKIH). To examine chelator efficacy, we assessed their effects on proliferation, Fe uptake, Fe efflux, the expression of cell cycle control molecules, iron regulatory protein-RNA-binding activity, and (3)H-thymidine, (3)H-uridine and (3)H-leucine incorporation. (2) Despite the high lipophilicity of the QCIH ligands and the fact that they have the same Fe-binding site as the PCIH series, surprisingly none of these compounds were effective. In contrast, the PKIH analogues showed marked anti-proliferative activity and Fe chelation efficacy. Indeed, the ability of these ligands to inhibit proliferation and DNA synthesis was similar or exceeded that found for the highly cytotoxic chelator, 311. In contrast to the PCIH and QCIH analogues, most of the PKIH group markedly increased the mRNA levels of molecules vital for cell cycle arrest. (3) In conclusion, our studies identify structural features useful in the design of chelators with high anti-proliferative activity. We have identified a novel class of ligands that are potent Fe chelators and inhibitors of DNA synthesis, and which deserve further investigation. PMID- 12642384 TI - Ocular hypotensive effects of melatonin receptor agonists in the rabbit: further evidence for an MT3 receptor. AB - (1) Melatonin is involved in the control of intraocular pressure during the night and day photoperiod. We have investigated the receptor that regulates intraocular pressure in New Zealand white rabbits by means of agonists and antagonists of melatonin receptors. (2) Melatonin and its analogues: 2-Phe-melatonin, 6-Cl melatonin, 2-I-melatonin, 5- methoxycarbonylamino-N-acetyltryptamine (5-MCA-NAT) and N-acetyltryptamine all produced a reduction in intraocular pressure. Dose response analysis for these compounds gave pD(2) values of 9.3+/-0.24 for melatonin; 9.0+/-0.09 for 6-Cl-melatonin; 9.0+/-0.84 for 2-I-melatonin; 8.9+/ 0.07 for 5-MCA-NAT; 8.7+/-0.18 for 2-Phe-melatonin and 9.4+/-0.30 for N acetyltryptamine (all n=8). (3) At a dose of 0.5 nmol (in 10 micro l) melatonin and the selective melatonin MT(3) agonist 5-MCA-NAT, induced greater reductions of intraocular pressure (22.8+/-2.3% and 32.5+/-1.4%, respectively) than the other compounds. (4) The melatonin-receptor antagonists, prazosin, DH-97 and 4-P PDOT, reversed the effect of 5-MCA-NAT in a dose-dependent manner, with pA(2) values of 13.5+/-0.17 for prazosin, 10.6+/-0.16 for DH-97 and 9.4+/-0.20 for 4-P PDOT (n=8). (5) Cholinoceptor antagonists (hexamethonium and atropine) and alpha(2)- and beta(2)-adrenoceptor antagonists (yohimbine and ICI 118,551) partially reversed the effects produced by melatonin and 5-MCA-NAT, suggesting the possible involvement of cholinergic and noradrenergic systems in the hypotensive actions mediated by melatonin agonists. The alpha(1)-adrenoceptor antagonist, corynanthine, had no significant effect. (6) The strong hypotensive effect of the MT(3) agonist, 5-MCA-NAT, suggests that this compound may be a useful agent for treating those pathologies where intraocular pressure is abnormally elevated. PMID- 12642385 TI - Mechanisms of endothelial dysfunction after ionized radiation: selective impairment of the nitric oxide component of endothelium-dependent vasodilation. AB - (1) Gamma radiation impairs vascular function, leading to the depression of endothelium-dependent vasodilatation. Loss of the nitric oxide (NO) pathway has been implicated, but little is known about radiation effects on other endothelial mediators. (2) This study investigated the mechanisms of endothelial dysfunction in rabbits subjected to whole-body irradiation from a cobalt(60) source. (3) The endothelium-dependent relaxation of rabbit aorta evoked by acetylcholine (ACh) or A23187 was impaired in a dose-dependent manner by irradiation at 2 Gy or above. Inhibition was evident 9 days post-irradiation and persisted over the 30 day experimental period. (4) Endothelium-independent responses to glyceryl trinitrate (GTN), sodium nitroprusside (SNP) and 3-morpholino-sydnonimine (SIN-1) were suppressed over a similar dose range at 7-9 days post-irradiation, but recovered fully by 30 days post-irradiation. (5) In healthy vessels, ACh-induced relaxation was inhibited by L-N(omega)-nitroarginine (L-NA; 3 x 10(-4) M) and charybdotoxin (10(-8) M) plus apamin (10(-6) M) but resistant to indomethacin, indicating the involvement of NO and endothelium-derived hyperpolarizing factor (EDHF). Supporting this, ACh caused smooth muscle hyperpolarization that was reduced by L NA and charybdotoxin plus apamin. (6) In irradiated vessels, responses to ACh were insensitive to L-NA but abolished by charybdotoxin plus apamin, indicating selective loss of NO-mediated relaxation. (7) In animals treated shortly after irradiation with the antioxidant, alpha-tocopherol acetate, the NO-dependent relaxation was restored without effect on the EDHF-dependent component. (8) The results imply that radiation selectively impairs the NO pathway as a consequence of oxidative stress, while EDHF is able to maintain endothelium-dependent relaxation at a reduced level. PMID- 12642387 TI - S-nitrosocaptopril: in vitro characterization of pulmonary vascular effects in rats. AB - (1) On rat isolated pulmonary arteries, vasorelaxation by S-nitrosocaptopril (SNOcap) was compared with S-nitrosoglutathione (GSNO) and nitroprusside, and inhibition by SNOcap of contractions to angiotensin I was compared with the angiotensin converting enzyme (ACE) inhibitor, captopril. (2) SNOcap was equipotent as a vasorelaxant on main (i.d. 2-3 mm) and intralobar (i.d. 600 micro m) pulmonary arteries (pIC(50) values: 5.00 and 4.85, respectively). Vasorelaxant responses reached equilibrium rapidly (2-3 min). (3) Pulmonary vasorelaxant responses to SNOcap, like GSNO, were (i) partially inhibited by the soluble guanylate cyclase inhibitor, ODQ (1H-(1,2,4) oxadiazolo(4,3-a)-quinoxalin-1-one; 3 micro M) whereas responses to nitroprusside were abolished and (ii) potentiated by hydroxocobalamin (HCOB; NO. free radical scavenger; 100 micro M) whereas responses to nitroprusside were inhibited. (4) The relative potencies for pulmonary vasorelaxation compared with inhibition of platelet aggregation were: SNOcap 7 : 1; GSNO 25 : 1; nitroprusside >2000 : 1. (5) SNOcap, like captopril, concentration-dependently and time-dependently increased the EC(50) for angiotensin I but not angiotensin II. The dependence on incubation time was independent of the presence of tissue but differed for SNOcap and captopril. This difference reflected the slow dissociation of SNOcap and instability of captopril, and precluded a valid comparison of the potency of the two drugs. After prolonged incubation (>/=5.6 h) SNOcap was more effective than captopril. (6) Thus, in pulmonary arteries SNOcap (i) possesses NO donor properties characteristic of S-nitrosothiols but different from nitroprusside and (ii) inhibits ACE at least as effectively as captopril. These properties suggest that SNOcap could be valuable in the treatment of pulmonary hypertension. PMID- 12642386 TI - RXR activators molecular signalling: involvement of a PPAR alpha-dependent pathway in the liver and kidney, evidence for an alternative pathway in the heart. AB - (1) In this study we compared the molecular signalling elicited by rexinoids, selective retinoid X receptor (RXR)-activators, in several organs (i.e. liver, kidney, heart) and in hepatocytes of various species. (2) RXR plays the pivotal role of a hetero-dimerization partner for the members of the class II subset of nuclear receptors which regulate the transcription of numerous target genes, following chemical activation. Several of these selective activators are currently used to treat hyperlipidaemia (fibrates), type II diabetes (glitazones), or skin disorders (retinoic acid). Although these therapeutic pathways are not fully elucidated, receptor activation is considered a pre requisite for efficacy. Therefore RXR, which accepts numerous dimeric partners, is considered a worthwhile pharmacological target. (3) We analysed a number of biochemical and molecular responses to rexinoids which were given orally to mice. Our results showed a prominent involvement of the peroxisome proliferator activated receptor (PPARalpha) as a majority of the observed hepatic and renal regulations were abolished in PPARalpha-knockout animals. Therefore we documented the species-specificity of these rexinoid actions which were reproduced in rat primary hepatocyte cultures but not in cultures of rabbit or human origin. Conversely, we established that the regulation of the pyruvate dehydrogenase kinase (PDK4) gene in the heart, by rexinoids, is independent of PPARalpha expression. (4) Our results support the obligatory expression of the active, although quiescent, PPARalpha to sustain a subset of relevant regulations attributable to rexinoids in the liver and kidney. Their cardiac molecular signalling unveiled an alternate transduction pathway and therefore opens new prospects in the therapeutic potential of rexinoids. PMID- 12642388 TI - Protease-activated receptor (PAR)-independent growth and pro-inflammatory actions of thrombin on human cultured airway smooth muscle. AB - (1) Thrombin, a mitogen for human cultured airway smooth muscle (HASM), has many actions that have been attributed to activation of protease-activated receptor (PARs). However, the role of PARs in the proliferative action has not been clearly identified. Moreover, thrombin elicits cytokine production in a number of cell types, but these effects have not been characterized in human ASM. (2) Thrombin (0.03-3 U ml(-1))-stimulated increases in the levels of the pro inflammatory and fibrogenic cytokine, granulocyte-macrophage colony-stimulating factor (GM-CSF) were observed over the same concentration range observed for thrombin-stimulated mitogenesis. (3) Inhibition of thrombin proteolytic activity, with either D-phenylalanyl-L-prolyl-L-arginine chloromethyl ketone (PPACK)- or hirudin-treated thrombin (0.3 U ml(-1)) or in the presence of the thrombin serine protease-selective inhibitor, SDZ 217-766 (0.15 micro M), reduced the thrombin stimulated GM-CSF levels by 91+/-3, 65+/-12 and 83+/-9% (n=8, P<0.05), respectively. PPACK treatment, hirudin and SDZ 217-766 inhibited thrombin stimulated increase in cell number by 70+/-8, 63+/-11 and 69+/-8%, respectively. (4) PAR-selective peptides SFLLRN (PAR1; 10 micro M), SLIGKV (PAR2; 10 micro M), GYPGQV (PAR4; 100 micro M) or the combination of SFLLRN and GYPGQV elicited mitogenic responses of only 15% of that to thrombin and surprisingly, had no effect on GM-CSF levels (n=8). Nevertheless, inhibition of thrombin responses by pertussis toxin (50 ng ml(-1)) suggests that the PAR-independent actions also involve a G-protein-coupled receptor. (5) PAR1 receptor expression was evident by immunohistochemistry and these receptors were coupled to increases in intracellular calcium, but not to the phosphorylation of ERK or the increases in cyclin D1 protein levels that are essential for cell proliferation. Cross desensitization of intracellular calcium increases by thrombin and the PAR1 selective peptide provides evidence that the PAR1 receptor responds to both ligands. (6) The failure of PAR-selective peptides to mimic thrombin responses together with the inhibition of thrombin responses by serine protease inhibitors suggest the involvement of novel proteolytic receptor targets for thrombin induced mitogenesis and cytokine production. PMID- 12642389 TI - Local anaesthetics have different mechanisms and sites of action at the recombinant N-methyl-D-aspartate (NMDA) receptors. AB - (1) Although the principal pharmacological targets of local anaesthetics (LAs) are voltage-gated Na(+) channels, other targets have also been suggested. Here we examined the effects of LAs on the N-methyl-D-aspartate (NMDA) receptor, a receptor involved in the process of nociception. (2) LAs (bupivacaine, lidocaine, procaine, and tetracaine) reversibly and concentration-dependently inhibited recombinant epsilon1/zeta1 and epsilon2/zeta1 NMDA receptors expressed in Xenopus oocytes (IC(50)s for bupivacaine, lidocaine, procaine, and tetracaine were 1032.0, 1174.1, 642.1 and 653.8 micro M at the epsilon1/zeta1 receptor; and 1090.8, 1821.3, 683.0 and 662.5 micro M respectively (at the epsilon2/zeta1 receptor). Bupivacaine and procaine were non-competitive antagonists; bupivacaine possesses non-competitive and competitive actions when interacting with glycine, whereas procaine has only non-competitive action. (3) Mutation of asparagine residue at position 598 (Asp(598)) in the zeta1 subunit, a residue associated with the blockade site for Mg(2+) and ketamine, to glutamine or arginine reduced the sensitivity to procaine but not to bupivacaine. Thus, procaine may interact with sites of action that are closely related to those of Mg(2+) and ketamine blockade. (4) These results suggest that LAs inhibit the NMDA receptor by various mechanisms. PMID- 12642390 TI - Pharmacological characterization of the homomeric and heteromeric UNC-49 GABA receptors in C. elegans. AB - (1) UNC-49B and UNC-49C are gamma-aminobutyric acid (GABA) receptor subunits encoded by the Caenorhabditis elegans unc-49 gene. UNC-49B forms a homomeric GABA receptor, or can co-assemble with UNC-49C to form a heteromeric receptor. The pharmacological properties of UNC-49B homomers and UNC-49B/C heteromers were investigated in Xenopus oocytes. (2) The UNC-49 subunits are most closely related to the bicuculline- and benzodiazepine-insensitive RDL GABA receptors of insects. Consistent with this classification, bicuculline (10 micro M) did not inhibit, nor did diazepam (10 micro M) enhance UNC-49B homomeric or UNC-49B/C heteromeric receptors. (3) The UNC-49C subunit strongly affects the pharmacology of UNC-49B/C heteromeric receptors. UNC-49B homomers were much more picrotoxin sensitive than UNC-49B/C heteromers (IC(50)=0.9+/-0.2 micro M and 166+/-42 micro M, respectively). Pentobarbitone enhancement was greater for UNC-49B homomers compared to UNC-49B/C heteromers. Propofol (50 micro M) slightly enhanced UNC-49B homomers but slightly inhibited UNC-49B/C heteromers. Penicillin G (10 mM) inhibited UNC-49B homomers less strongly than UNC-49B/C heteromers (30% compared to 53% inhibition, respectively). (4) Several aspects of UNC-49 pharmacology were unusual. Picrotoxin sensitivity strongly correlates with dieldrin sensitivity, yet UNC-49B homomers were highly dieldrin resistant. The enhancing neurosteroid pregnanolone (5beta-pregnan-3alpha-ol-20-one; 10 micro M) strongly inhibited both UNC-49 receptors. Alphaxalone (10 micro M), another enhancing neurosteroid, did not affect UNC-49B homomers, but slightly inhibited UNC-49B/C heteromers. (5) UNC 49 subunits and mammalian GABA(A) receptor alpha, beta, and gamma subunit classes all share roughly the same degree of sequence similarity. Thus, although they are most similar to other invertebrate GABA receptors, the UNC-49 receptors share significant structural and pharmacological overlap with mammalian GABA(A) receptors. PMID- 12642391 TI - Role of nuclear factor-kappa B activation in acute ischaemia-reperfusion injury in myocardium. AB - (1) Our aims were to characterize activation of the transcription factor, nuclear factor kappa-B(NF-kappaB), during myocardial ischaemia-reperfusion and to assess its functional role in the evolution of acute ischaemia-reperfusion injury in intact myocardium in vivo. (2) Under pentobarbitone anaesthesia, rabbits underwent sham operation, 30 min left coronary artery occlusion followed by 0, 10 or 180 min reperfusion. Saline or NF-kappaB inhibitor diethyldithiocarbamic acid (DDTC, 50, 100 or 200 mg kg(-1)) was given intravenously 5 min prior to reperfusion. (3) Electromobility shift assay revealed that 30 min ischaemia alone did not activate NF-kappaB compared to time-matched sham-operated controls (85+/ 13% vs 100+/-28%, respectively). However, ischaemia plus 10 min reperfusion markedly increased activation of NF-kappaB (295+/-77%). DDTC 50 mg kg(-1) did not inhibit NF-kappaB activation (278+/-67%) but at the higher doses complete inhibition was observed (54+/-20%, 31+/-16%, respectively). (4) Infarct to risk ratio was determined by triphenyltetrazolium chloride staining after 30 min ischaemia and 180 min reperfusion. DDTC 50 or 100 mg kg(-1) significantly reduced infarct size compared to the saline-treated control group (34.9+/-5.2%, 37.1+/ 5.9%, vs 51.3+/-3.6%, P<0.05, respectively), whereas there was no protection with 200 mg kg(-1) (45.6+/-5.3%). (5) We conclude that ischaemia alone does not activate NF-kappaB, but post-ischaemic reperfusion robustly activates NF-kappaB in the myocardium. DDTC limited irreversible injury at low doses, but this effect appears to be dissociated from inhibition of NF-kappaB. Thus, activation of NF kappaB during reperfusion does not appear to play a role in the evolution of myocardial infarction during the early phase of reperfusion. PMID- 12642392 TI - Apoptosis and necrosis: two distinct events induced by cadmium in cortical neurons in culture. AB - (1) Cadmium is an extremely toxic metal commonly found in industrial workplaces, a food contaminant and a major component of cigarette smoke. Cadmium can severely damage several organs, including the brain. In this work, we have studied both the cadmium toxicity on rat cortical neurons in culture and the possible protective effect of serum. (2) Our results indicate that: (1) cadmium is taken up by the neurons in a dose and serum dependent way; (2) cadmium, at concentrations from 1 micro M or 10 micro M (depending on the absence or the presence of serum) up to 100 micro M, decreases the metabolic capacity, which was evaluated by the XTT (tetrazolium salt) test; (3) cadmium induces apoptosis and LDH (lactate dehydrogenase) release in a dose dependent way; (4) in a serum-free medium, the cadmium-induced apoptosis is accompanied by caspase-3 activation; (5) both the caspase-3 activation and the cadmium-induced apoptosis are reversed by N acethyl-Asp-Glu-Val-Asp-aldehyde (Ac-DEVD-CHO), a selective caspase-3 inhibitor, indicating that the caspase-3 pathway is involved in cadmium-induced apoptosis in cortical neurons; and (6) the cadmium concentrations which produce caspase-3 activation do not modify the intracellular ATP levels; however, higher cadmium concentrations lead to both intracellular ATP depletion and ATP release, but do not increase the caspase-3 activity, indicating that cadmium also produces cellular death by necrosis. (3) These results suggest that cadmium induces either apoptosis or necrosis in rat cortical neurons, depending on the cadmium concentration. PMID- 12642393 TI - Role of Th2 responses in the development of allergen-induced airway remodelling in a murine model of allergic asthma. AB - (1) To clarify the involvement of Th2 responses in the development of allergen induced airway remodelling, we investigated the effect of anti-CD4 monoclonal antibody (mAb) and anti-CD8 mAb, and the responses of IL-4 gene-knockout (KO) mice in a murine model of allergic asthma. (2) Mice were immunized twice by intraperitoneal injections of ovalbumin (OA), and exposed to aeroallergen (OA, 1% w v(-1)) for 3 weeks. Twenty-four hours after the final challenge, airway responsiveness to acetylcholine was measured, and bronchoalveolar lavage (BAL) and histological examinations were carried out. (3) Anti-CD4 mAb (1 mg kg(-1)) clearly inhibited allergen-induced increases in airway responsiveness to acetylcholine, the number of eosinophils in BAL fluid, serum OA-specific IgE levels, IL-13 and transforming growth factor-beta1 levels in BAL fluid, and amount of hydroxyproline in the lung by 100, 99, 100, 100, 84, and 60%, respectively. Furthermore, the antibody (1 mg kg(-1)) also attenuated allergen induced goblet cell hyperplasia in the epithelium and subepithelial fibrosis by 72 and 83%, respectively. In contrast, anti-CD8 mAb (1 mg kg(-1)) showed no effect on each parameter. Furthermore, all these parameters were attenuated in IL 4KO mice by 57, 93, 100, 45, 84 and 60%, and also 72 and 83%, respectively. (4) These findings suggest that Th2 responses play a critical role for the development of allergen-induced airway remodelling, and that the inhibition of Th2 responses, e.g. using anti-CD4 mAb, is a therapeutic approach for the treatment of airway remodelling in asthma. PMID- 12642394 TI - Desensitization of alpha 2A-adrenoceptor signalling by modest levels of adrenaline is facilitated by beta 2-adrenoceptor-dependent GRK3 up-regulation. AB - (1) Adrenaline (ADR) and noradrenaline (NA) can simultaneously activate inhibitory alpha(2)- and stimulatory beta-adrenoceptors (AR). However, ADR and NA differ significantly in that ADR is a potent beta(2)-AR agonist while NA is not. Only recently has the interaction resulting from the simultaneous activation of alpha(2)- and beta(2)-AR been examined at the cellular level to determine the mechanisms of alpha(2)-AR regulation following concomitant activation of both alpha(2)- and beta(2)-ARs by chronic ADR. (2) This study evaluates beta(2)-AR regulation of alpha(2A)-AR signalling following chronic ADR (300 nM) and NA (1 and 30 micro M) treatments of BE(2)-C human neuroblastoma cells that natively express both beta(2)- and alpha(2A)-ARs. (3) Chronic (24 h) treatment with ADR (300 nM) desensitized the response to the alpha(2A)-AR agonist, brimonidine, in BE(2)-C cells. Addition of the beta-AR antagonist, propranolol, blocked the ADR induced alpha(2A)-AR desensitization. Unlike ADR, chronic NA (1 micro M) treatment had no effect on the alpha(2A)-AR response. However if NA was increased to 30 micro M for 24 h, alpha(2A)-AR desensitization was observed; this desensitization was partially reversed by propranolol. (4) Chronic ADR (300 nM) treatment reduced alpha(2A)-AR binding levels, contributing to the alpha(2A)-AR desensitization. This decrease was prevented by addition of propranolol during ADR treatment. Chronic NA (30 micro M), like ADR, treatment lowered specific binding, whereas 1 micro M NA treatment was without effect. (5) Chronic ADR treatment produced a significant increase in GRK3 levels and this was blocked by propranolol or GRK2/3 antisense DNA treatment. This antisense DNA, common to both GRK2 and GRK3, also blocked chronic ADR-induced alpha(2A)-AR desensitization and down-regulation. (6) Acute (1 h) ADR (300 nM) or NA treatment (1 micro M) produced alpha(2A)-AR desensitization. The desensitization produced by acute treatment was beta-AR independent, as it was not blocked by propranolol. (7) We conclude that chronic treatment with modest levels of ADR produces alpha(2A)-AR desensitization by mechanisms that involve up-regulation of GRK3 and down regulation of alpha(2A)-AR levels through interactions with the beta(2)-AR. PMID- 12642395 TI - A photosensitive vascular smooth muscle store of nitric oxide in mouse aorta: no dependence on expression of endothelial nitric oxide synthase. AB - (1) Photorelaxation is the reversible relaxation of vascular smooth muscle (VSM) when irradiated with ultraviolet (UV) light resulting from the release of nitric oxide (NO). In this study we characterize the involvement of endothelial nitric oxide synthase (eNOS) in the photorelaxation response of thoracic aorta from endothelial NOS deficient (-/-) and control (C57BL/6j) mice. (2) Cirazoline contracted aortae were repeatedly exposed to 30 s of UV light every 3-4 min. Equal levels of photorelaxation (45+/-2%; n=34) was observed in both strains. (3) 1H-[1,2,4]-oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), K(+), 2-(4-carboxyphenyl) 4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO), 4-aminopyridine (4-AP) and ethacrynic acid significantly reduced the photorelaxation response. In C57BL/6j mice diethyldithiocarbamate (DETCA) also reduced photorelaxation. (4) Control endothelium-intact and -denuded aorta and L-NAME (100 micro M) treated and untreated eNOS (-/-) aortae were repeatedly exposed to UV light for 5 min every 10 min until no photorelaxation response was observed. After 1 h of rest in the dark the vessels showed between 30-70% recovery of the photorelaxation response indicating regeneration of the store in the absence of the endothelium and eNOS. (5) The results of this study suggest that photorelaxation in mouse aorta VSM results from the release of NO from a stable store of RSNOs, which activates soluble guanylate cyclase (sGC), leading to cGMP-dependent relaxation that is partially mediated by an increase in K(V) channel activation and hyperpolarization. In addition, the eNOS isoform is not essential for the formation of the photorelaxation store and a non-NOS source of NO may be involved in the maintenance of this store. PMID- 12642396 TI - Anti-amnesic effect of dimemorfan in mice. AB - (1) Dimemorfan, an antitussive for more than 25 years, has previously been reported to be a relative high-affinity ligand at sigma-1 (sigma(1)) receptor with the K(i) value of 151 nM. (2) To test whether dimemorfan has anti-amnesic effects similar to a sigma(1) receptor agonist, this study examined its effects on scopolamine- and beta-amyloid peptide-(25-35)-induced amnesia in mice. (3) Dimemorfan (10-40 mg kg(-1), i.p.) administered 30 min before the training trial, immediately after the training trial, or 30 min before the retention test significantly improved scopolamine (1 mg kg(-1), i.p.)- or beta-amyloid peptide (25-35) (3 nmol mouse(-1), i.c.v.)-induced amnesia in a step-through passive avoidance test. Dimemorfan (5-40 mg kg(-1), i.p.) pretreatment also attenuated scopolamine (8 mg kg(-1), i.p.)-induced amnesia in a water-maze test. And, these anti-amnesic effects of dimemorfan, like the putative sigma(1) receptor agonist (+)-N-allylnormetazocine ((+)-SKF-10047), were antagonized by a sigma receptor antagonist haloperidol (0.25 mg kg(-1), i.p.). (4) These results indicated that dimemorfan has anti-amnesic effects and acts like a sigma(1) receptor agonist. PMID- 12642397 TI - Inhibition by pentoxifylline of TNF-alpha-stimulated fractalkine production in vascular smooth muscle cells: evidence for mediation by NF-kappa B down regulation. AB - (1) Fractalkine is a CX(3)C chemokine for mononuclear leukocytes that is expressed mainly by vascular cells, and regulated by pro-inflammatory cytokines. This study investigated signal transduction mechanisms by which tumor necrosis factor (TNF)-alpha stimulated fractalkine expression in cultured rat vascular smooth muscle cells (VSMCs), and the modulatory effect of a haemorrheologic agent, pentoxifylline, on its production. (2) TNF-alpha (1-50 ng ml(-1)) stimulated fractalkine mRNA and protein expression in concentration- and time dependent manners. Pretreatment with calphostin C (0.4 micro M, a selective inhibitor of protein kinase C (PKC), and PD98059 (40 micro M), a specific inhibitor of p42/44 mitogen-activated protein kinase (MAPK) kinase, attenuated TNF-alpha-stimulated fractalkine mRNA and protein expression. In contrast, H-89 (2 micro M), a selective inhibitor of cAMP-dependent protein kinase, wortmannin (0.5 micro M), a selective inhibitor of phosphatidylinositol 3-kinase, and SB203580 (40 micro M), a specific inhibitor of p38 MAPK, had no discernible effect. (3) The ubiquitin/proteosome inhibitors, MG132 (10 micro M) and pyrrolidine dithiocarbamate (200 micro M), suppressed activation of NF-kappaB as well as stimulation of fractalkine mRNA and protein expression by TNF-alpha. (4) TNF-alpha-activated phosphorylation of PKC was blocked by calphostin C, whereas TNF-alpha-augmented phospho-p42/44 MAPK and phospho-c-Jun levels were reduced by PD98059. Neither calphostin C nor PD98059 affected TNF-alpha-induced degradation of I-kappaBalpha or p65 nuclear translocation. (5) Pretreatment with pentoxifylline (0.1-1 mg ml(-1)) decreased TNF-alpha-stimulated fractalkine mRNA and protein expression, which was preceded by a reduction in TNF-alpha-activated phosphorylation of PKC, p42/44 MAPK and c-Jun as well as degradation of I kappaBalpha and p65/NF-kappaB nuclear translocation. (6) These data indicate that activation of PKC, p42/44 MAPK kinase, and NF-kappaB are involved in TNF-alpha stimulated fractalkine production in VSMCs. Down-regulation of the PKC, p42/44 MAPK, and p65/NF-kappaB signals by PTX may be therapeutically relevant and provide an explanation for the anti-fractalkine effect of this drug. PMID- 12642398 TI - A potent tryptase inhibitor nafamostat mesilate dramatically suppressed pulmonary dysfunction induced in rats by a radiographic contrast medium. AB - (1) Intravenous injection of ioxaglate (4 g iodine kg(-1)), an iodinated radiographic contrast medium, caused a marked protein extravasation, pulmonary oedema and a decrease in the arterial partial oxygen pressure in rats. (2) All of these reactions to ioxaglate were reversed by the pretreatment with gabexate mesilate (10 and 50 mg kg(-1), 5 min prior to injection) or nafamostat mesilate (3 and 10 mg kg(-1)), in which the inhibition was complete after injection of nafamostat mesilate (10 mg kg(-1)). (3) Both gabexate mesilate and nafamostat mesilate inhibited the activity of purified human lung tryptase, although the latter compound was far more potent than the former. (4) Ioxaglate enhanced the nafamostat-sensitive protease activity in the extracellular fluid of rat peritoneal mast cell suspensions. (5) Tryptase enhanced the permeability of protein through the monolayer of cultured human pulmonary arterial endothelial cells. Ioxaglate, when applied in combination with rat peritoneal mast cells, also produced the endothelial barrier dysfunction. These effects of tryptase and ioxaglate were reversed by nafamostat mesilate. (6) Consistent with these findings, immunofluorescence morphological analysis revealed that tryptase or ioxaglate in combination with mast cells increased actin stress fibre formation while decreasing VE-cadherin immunoreactivity. Both of these actions of tryptase and ioxaglate were reversed by nafamostat mesilate. (7) These findings suggest that tryptase liberated from mast cells plays a crucial role in the ioxaglate induced pulmonary dysfunction. In this respect, nafamostat mesilate may become a useful agent for the cure or prevention of severe adverse reactions to radiographic contrast media. PMID- 12642399 TI - Dopamine D2-like receptor-mediated opening of K+ channels in opossum kidney cells. AB - (1) This study examined the effects of dopamine D(1)- and D(2)-like receptor activation upon basolateral K(+) (I(K)) currents and changes in membrane potential in opossum kidney (OK) cells. (2) The addition of amphotericin B (3 micro g ml(-1)) to the apical side resulted in a rapid increase in I(K), this effect being markedly inhibited by the addition of the K(+) channel blockers barium chloride (1 mM) or glibenclamide (10 micro M), but not apamin (1 micro M). The K(+) channel opener pinacidil increased the amphotericin B-induced I(K). The selective D(2)-like receptor agonist quinerolane increased, in a concentration dependent manner (EC(50)=136 nM), I(K) across the basolateral membrane, this effect being abolished by pre-treatment with pertussis toxin (PTX), S-sulpiride (selective D(2)-like receptor antagonist) and glibenclamide. The selective D(1) like receptor agonist SKF 38393 did not change I(K). Both H-89 (PKA inhibitor) and chelerythrine (PKC inhibitor) failed to prevent the stimulatory effect of quinerolane upon I(K). (3) Quinerolane did not change basal levels of cyclic AMP and also failed to affect the forskolin-induced increase in cyclic AMP levels. (4) The stimulation of D(2)-like receptor was associated with a rapid hyperpolarizing effect, whereas D(1)-like receptor activation was accompanied by increases in cell membrane potential. The hyperpolarizing effect of quinerolane (EC(50)=129 nM) was prevented by pre-treatment with PTX, S-sulpiride and glibenclamide. (5) It is concluded that stimulation of dopamine D(2)-like, but not D(1)-like, receptors coupled to PTX-sensitive G proteins of the G(i/o) class produce membrane hyperpolarization through opening of K(ATP) channels. PMID- 12642400 TI - Neurogenic responses mediated by vanilloid receptor-1 (TRPV1) are blocked by the high affinity antagonist, iodo-resiniferatoxin. AB - (1) Stimulation of the vanilloid receptor-1 (TRPV1) results in the activation of nociceptive and neurogenic inflammatory responses. Poor specificity and potency of TRPV1 antagonists has, however, limited the clarification of the physiological role of TRPV1. (2) Recently, iodo-resiniferatoxin (I-RTX) has been reported to bind as a high affinity antagonist at the native and heterologously expressed rat TRPV1. Here we have studied the ability of I-RTX to block a series of TRPV1 mediated nociceptive and neurogenic inflammatory responses in different species (including transfected human TRPV1). (3) We have demonstrated that I-RTX inhibited capsaicin-induced mobilization of intracellular Ca(2+) in rat trigeminal neurons (IC(50) 0.87 nM) and in HEK293 cells transfected with the human TRPV1 (IC(50) 0.071 nM). (4) Furthermore, I-RTX significantly inhibited both capsaicin-induced CGRP release from slices of rat dorsal spinal cord (IC(50) 0.27 nM) and contraction of isolated guinea-pig and rat urinary bladder (pK(B) of 10.68 and 9.63, respectively), whilst I-RTX failed to alter the response to high KCl or SP. (5) Finally, in vivo I-RTX significantly inhibited acetic acid-induced writhing in mice (ED(50) 0.42 micro mol kg(-1)) and plasma extravasation in mouse urinary bladder (ED(50) 0.41 micro mol kg(-1)). (6) In in vitro and in vivo TRPV1 activated responses I-RTX was approximately 3 log units and approximately 20 times more potent than capsazepine, respectively. This high affinity antagonist, I-RTX, may be an important tool for future studies in pain and neurogenic inflammatory models. PMID- 12642401 TI - Inhibition of cyclic GMP hydrolysis with zaprinast reduces basal and cyclic AMP elevated L-type calcium current in guinea-pig ventricular myocytes. AB - (1) Cyclic GMP (cGMP) has been shown to be an important modulator of cardiac contractile function. A major component of cGMP regulation of contractility is cGMP-mediated inhibition of the cardiac calcium current (I(Ca)). An under appreciated aspect of cyclic nucleotide signalling is hydrolysis of the cyclic nucleotide (i.e., breakdown by phosphodiesterases (PDEs)). The role of cGMP hydrolysis in regulating I(Ca) has not been studied. Thus the purpose of this study was to investigate if inhibition of cGMP hydrolysis can modulate I(Ca) in isolated guinea-pig ventricular myocytes. (2) Zaprinast, a selective inhibitor of cGMP-specific PDE (PDE5), caused a significant increase in cGMP levels in myocytes, but was without affect on basal or beta-adrenergic stimulated cAMP levels (consistent with its actions as a specific inhibitor of PDE5). (3) Zaprinast inhibited I(Ca) that was pre-stimulated with cAMP elevating agents (isoproterenol, a beta-adrenergic agonist; or forskolin, a direct activator of adenylate cyclase). The effect of zaprinast was greatly reduced by KT5823, an inhibitor of cGMP-dependent protein kinase (PKG). (4) Zaprinast also significantly inhibited basal I(Ca) when perforated-patch or whole-cell recording with physiological pipette calcium concentration (10(-7) M) was used. However, this effect was not observed when using standard calcium-free whole-cell recording conditions. (5) These results indicate that inhibition of cGMP hydrolysis can decrease both basal and cAMP-stimulated I(Ca). Thus, cGMP hydrolysis may likely be an important step for physiological modulation of I(Ca). This regulation may also be important in disease states in which cGMP production is increased and PDE5 expression is altered, such as heart failure. PMID- 12642402 TI - Regulation of intracellular cyclic AMP in skeletal muscle cells involves the efflux of cyclic nucleotide to the extracellular compartment. AB - (1) This report analyses the intracellular and extracellular accumulation of cyclic AMP in primary rat skeletal muscle cultures, after direct and receptor dependent stimulation of adenylyl cyclase (AC). (2) Isoprenaline, calcitonin gene related peptide (CGRP) and forskolin induced a transient increase in the intracellular cyclic AMP that peaked 5 min after onset stimulation. (3) Under stimulation with isoprenaline or CGRP, the intracellular cyclic AMP initial rise was followed by an exponential decline, reaching 46 and 52% of peak levels in 10 min, respectively. (4) Conversely, the forskolin-dependent accumulation of intracellular cyclic AMP decreased slowly and linearly, reaching 49% of the peak level in 30 min. (5) The loss of intracellular cyclic AMP from peak levels, induced by direct or receptor-induced activation of AC, was followed by an increase in the extracellular cyclic AMP. (6) This effect was independent on PDEs, since it was obtained in the presence of 3-isobutyl-1-methylxanthine (IBMX). (7) Besides, in isoprenaline treated cells, the beta-adrenoceptor antagonist propranolol reduced both intra- and extracellular accumulation of cyclic AMP, whereas the organic anion transporter inhibitor probenecid reduced exclusively the extracellular accumulation. (8) Together our data show that direct or receptor-dependent activation of skeletal muscle AC results in a transient increase in the intracellular cyclic AMP, despite the continuous presence of the stimulus. The temporal declining of intracellular cyclic AMP was not dependent on the cyclic AMP breakdown but associated to the efflux of cyclic nucleotide to the extracellular compartment, by an active transport since it was prevented by probenecid. PMID- 12642403 TI - The immunophilin-ligands FK506 and V-10,367 mediate neuroprotection by the heat shock response. AB - (1) The macrolid FK506 is widely used in transplantation to suppress allograft rejection. FK506 and its derivatives are powerful neuroprotective molecules, but the underlying mechanisms remain to be resolved. We have previously shown that the FK506 mediated neuroprotection against oxygen radicals is independent of the inhibition of calcineurin but depends on de novo protein synthesis. (2) Here, we have shown that FK506 mediates protection against H(2)O(2), UV-light or thapsigargin in neuronal cell lines, but not in non-neuronal cells such as R3T3 fibroblasts. We compared in detail the effect of FK506 on apoptotic features in PC12 cells after H(2)O(2) with V-10,367 which binds to FKBPs but does not inhibit calcineurin. Both molecules exert the same neuroprotective effect after H(2)O(2) stimulation. FK506, but not V-10,367, inhibited the cytochrome c release out of the mitochondria and the caspase 3 activation, while both molecules inhibited the cleavage of Poly-(ADP-ribose)-polymerase (Parp) and prevented the expression of p53. (3) FK506 and V-10,367 rapidly induced the expression of Hsp70 and Hsp27, but not Hsp90. Their neuroprotective actions could be completely blocked by quercetin, a functional inhibitor of the heat shock proteins. (4) We conclude that immunophilin-ligands such as FK506 and V-10,367 exert their neuroprotection independent of calcineurin through the induction of the heat shock response. The identification of the underlying signal transduction from application of immunophilin ligands to the expression of heat shock proteins represents a novel target cascade for neuroprotection. PMID- 12642404 TI - Expression and function of P2X purinoceptors in rat histaminergic neurons. AB - (1) The pharmacology of ATP responses and the expression pattern of seven known subunits of the P2X receptor were investigated in individual histaminergic neurons of the tuberomamillary nucleus (TM). (2) ATP (3-1000 micro M) evoked fast non-desensitizing inward currents in TM neurons. 2-methylthioATP (2MeSATP) displayed the same efficacy but a lower potency, EC(50)s 84 micro M versus 48 micro M, when compared with ATP. Adenosine-diphosphate (ADP), uridine triphosphate (UTP) and alpha beta methylene-ATP (alphabeta-meATP) were inactive. (3) ATP-mediated whole cell currents were potentiated by acidification of the recording solution (pH 7.5 and 6.6 were compared). (4) Single-cell RT-PCR (scRT PCR) analysis revealed that the P2X(2) receptor is expressed in all PCR-positive neurons. Each of the P2X(1), P2X(3), P2X(4), P2X(5) and P2X(6) mRNAs were detected in less than 35% of the cells. (5) Suramin antagonized ATP responses with an IC(50) of 4.2 micro M and pyridoxalphosphate-6-azophenyl-2',4' disulphonic acid (PPADS, 1 micro M) reduced ATP responses to 43% of control, when antagonists were pre-applied 90s before the agonist. Cibacron blue (3 micro M) given together with ATP potentiated control responses by 67%, but inhibited it to 10% after pre-application. (6) 2',3'-O-(2,4,6-Trinitrophenyl) adenosine 5' triphosphate (TNP-ATP) antagonized ATP responses with an IC(50) of 7 micro M. (7) Pharmacological properties of ATP responses together with scRT-PCR data suggest that P2X(2) is the major purinoceptor on the soma of TM neurons, however the presence of heteromeric P2X(2/5) receptors in some neurons cannot be excluded. PMID- 12642405 TI - Health in the hinterland. PMID- 12642411 TI - A Canadian history lesson. PMID- 12642408 TI - Propofol syndrome in children. PMID- 12642409 TI - Propofol syndrome in children. PMID- 12642412 TI - Snowmobiler's hematuria. PMID- 12642413 TI - Mandatory pharmacovigilance. PMID- 12642415 TI - CRA endorsement of osteoporosis guidelines. PMID- 12642416 TI - Responsibility in advertising. PMID- 12642417 TI - Antipsychotic drugs and diabetes. PMID- 12642419 TI - Osteoporosis in children: 2002 guidelines do not apply. PMID- 12642422 TI - Mandatory reporting of HIV infection and opt-out prenatal screening for HIV infection: effect on testing rates. AB - BACKGROUND: Mandatory reporting of HIV infection to public health authorities, although now common, may deter people from undergoing testing. We examined HIV testing frequency in Alberta before and after mandatory reporting was implemented. We also examined the effect on testing rates among pregnant women when Alberta adopted an opt-out approach to prenatal HIV screening. METHODS: Using data from the Provincial Laboratory for Public Health, we determined the number of HIV tests done between Jan. 1, 1993, and Dec. 31, 2000, for males and females in Alberta. We used data from the Canadian Blood Services laboratories to obtain the number of tests conducted as part of the opt-out prenatal HIV testing program. Reporting of HIV infection became mandatory on May 1, 1998, and opt-out prenatal HIV testing was introduced on Sept. 1, 1998. RESULTS: Among males, the average annual percent increase in the number of HIV tests was 4.0% for the period before mandatory testing, as compared with 4.3% for the period after mandatory reporting was implemented; the difference in yearly trend was significant (p < 0.001). Among females, the average annual percent increase in the number of HIV tests was 9.2% for the period before mandatory reporting. In the month immediately following the adoption of opt-out prenatal HIV testing, the rate increased by 28%. Between 1999 and 2000, the average annual percent increase in the number of HIV tests among females was 1.4%. INTERPRETATION: The introduction of mandatory reporting of HIV infection did not appear to have a deterrent effect on rates of HIV testing. The implementation of an opt-out prenatal HIV testing policy resulted in a dramatic increase in the number of females being tested for HIV infection. PMID- 12642423 TI - Do low-income lone mothers compromise their nutrition to feed their children? AB - BACKGROUND: Women who live in disadvantaged circumstances in Canada exhibit dietary intakes below recommended levels, but their children often do not. One reason for this difference may be that mothers modify their own food intake to spare their children nutritional deprivation. The objective of our study was to document whether or not low-income lone mothers compromise their own diets to feed their children. METHODS: We studied 141 low-income lone mothers with at least 2 children under the age of 14 years who lived in Atlantic Canada. Women were identified through community organizations using a variety of recruitment strategies. The women were asked weekly for 1 month to recall their food intake over the previous 24 hours; they also reported their children's (n = 333) food intake. Mothers also completed a questionnaire about "food insecurity," that is, a lack of access to adequate, nutritious food through socially acceptable means, during each interview. RESULTS: Household food insecurity was reported by 78% of mothers during the study month. Mothers' dietary intakes and the adequacy of intake were consistently poorer than their children's intake overall and over the course of a month. The difference in adequacy of intake between mothers and children widened from Time 1, when the family had the most money to purchase food, to Time 4, when the family had the least money. The children experienced some improvement in nutritional intake at Time 3, which was possibly related to food purchases for them associated with receipt of the Child Tax Benefit Credit or the Goods and Services Tax Credit. INTERPRETATION: Our study demonstrates that low-income lone mothers compromise their own nutritional intake in order to preserve the adequacy of their children's diets. PMID- 12642424 TI - Knowledge and attitudes of pregnant women with regard to collection, testing and banking of cord blood stem cells. AB - BACKGROUND: Umbilical cord blood is used as a source of hematopoietic stem cells for bone marrow transplantation in the treatment of malignant and nonmalignant disease. We sought to examine pregnant women's knowledge and attitudes regarding cord blood banking, as their support is crucial to the success of cord blood transplant programs. METHODS: A questionnaire examining sociodemographic factors and women's attitudes to cord blood banking was developed on the basis of findings from 2 focus groups and a pilot study. The questionnaire was distributed to 650 women attending antenatal clinics at a regional women's hospital between April and July 2001. RESULTS: A total of 443 women (68%) responded. More than half of the women (307/438 or 70% [95% confidence interval, CI, 66% to 74%]) reported poor or very poor knowledge about cord blood banking. Many of the respondents (299/441 or 68% [95% CI 63% to 72%]) thought that physicians should talk to pregnant women about the collection of cord blood, and they wanted to receive information about this topic from health care professionals (290/441 or 66% [95% CI 61% to 70%]) or prenatal classes (308/441 or 70% [95% CI 65% to 74%]). Most of the women (379/442 or 86% [95% CI 82% to 89%]) would elect to store cord blood in a public bank, many citing altruism as the reason for this choice. A much smaller proportion (63/442 or 14% [95% CI 11% to 18%]) would elect private banking, indicating that this would be a good investment or that they would feel guilty if the blood had not been stored. Additional acceptable uses for cord blood included research (mentioned by 294/436 women or 67% [95% CI 63% to 72%]) and gene therapy (mentioned by 169/437 women or 39% [95% CI 34% to 43%]). INTERPRETATION: Most of the women in this study supported the donation of cord blood to public cord blood banks for potential transplantation and research. PMID- 12642425 TI - Impact of universal preadolescent vaccination against hepatitis B on antenatal seroprevalence of hepatitis B markers in British Columbia women. AB - Countries with a low risk of hepatitis B (HB) lack data on the effectiveness of universal HB vaccination programs for children. British Columbia began a program in 1992, offering HB vaccination to 11 year olds. We conducted an anonymous, unlinked serologic survey 7 years later, analyzing a random sample of specimens (n = 1215) from women aged 15-44 years who had undergone antenatal rubella testing. Among those aged 15-19 years inclusive there was no evidence of chronic HB (HB surface antigen), the proportion with evidence of acute HB (anti-HB core antibody) was only 0.6% (compared with 6.5% for the entire sample), and evidence of protective immunity was strong: the prevalence of anti-HB surface antibody (anti-HBs) was 79.1% (compared with 41.4% for the entire sample) and the geometric mean titre was 34.9 IU/mL (compared with 0.6-0.8 IU/mL for the older groups [p < 0.001]). PMID- 12642426 TI - Opt in or opt out: what is optimal for prenatal screening for HIV infection? PMID- 12642427 TI - Low income, welfare and nutritional vulnerability. PMID- 12642428 TI - Technology-enabled knowledge translation: building a framework for collaboration. PMID- 12642429 TI - Assessment and management of attention-deficit hyperactivity disorder in adults. AB - Attention-deficit hyperactivity disorder (ADHD) is estimated to affect 2%-6% of adults. The symptoms in adults with ADHD mirror those in children with the disorder and are associated with significant educational, occupational and interpersonal difficulties. Double-blind, placebo-controlled trials have established that adult ADHD is responsive to stimulant medication treatment. New medications and psychotherapeutic approaches are being developed in an effort to achieve optimal treatment effects in this population. We review the available literature and provide an approach to the assessment and management of ADHD in adults. PMID- 12642431 TI - Salmeterol (Serevent) asthma trial halted early. PMID- 12642430 TI - "Palm reading": 2. Handheld software for physicians. PMID- 12642432 TI - Snowmobile injuries. PMID- 12642433 TI - Dexamethasone in adults with bacterial meningitis. PMID- 12642434 TI - Spontaneous paracentesis through an umbilical hernia. PMID- 12642439 TI - Medicine, the unhappy profession? PMID- 12642444 TI - New data point to significant gaps in income of male, female MDs. PMID- 12642457 TI - Cytochrome P450 in vitro reaction phenotyping: a re-evaluation of approaches used for P450 isoform identification. AB - Marker substrates, chemical inhibitors, and inhibitory antibodies are important tools for the identification of cytochrome P450 (P450) isoform responsible for the metabolism of therapeutic agents in vitro. In view of the versatile and nonspecific nature of P450 enzymes, many of the marker substrates and chemical inhibitors used for P450 in vitro reaction phenotyping are isoform selective but not specific. Recently, the use of marker substrate and chemical inhibitors in CYP2D6 in vitro reaction phenotyping was questioned by Granvil et al. (2002). In comparison of a panel of 15 recombinant P450 enzymes, they found that in addition to CYP2D6, CYP1A1 is also capable of catalyzing the formation of 4-hydroxylated metabolite of debrisoquine and that the intrinsic clearance of debrisoquine by CYP2D6-mediated 4-hydroxylation is only about twice that by CYP1A1. In their study, they have also demonstrated that quinidine inhibits both CYP2D6- and CYP1A1-mediated debrisoquine 4-hydroxylation. In view of these important findings, we have reevaluated various approaches used to identify P450 isoform(s) responsible for the metabolism of therapeutic agents. While acknowledging the value of inhibitory antibodies in P450-phenotyping studies, it is our opinion that in well conducted in vitro experiments, isoform-selective chemical inhibitors can also provide valuable and reliable information. Hopefully, future efforts may produce even better P450 isoform-selective marker substrates and inhibitors. PMID- 12642446 TI - "Floodgates of hope" open for HIV-ravaged nations. PMID- 12642458 TI - Direct identification of cytochrome P450 isozymes by matrix-assisted laser desorption/ionization time of flight-based proteomic approach. AB - The main targets of our investigation were cytochrome P450 isozymes (P450), the key enzymes of the hepatic drug-metabolizing system. Current research approaches to the identification of individual P450 forms include specific P450 inhibitors or substrates, antibody-based identification, and mRNA-based expression profiling. All of these approaches suffer from one common disadvantage-they all are indirect methods. On the other hand, current developments in mass spectrometry provide a direct and reliable approach to protein identification with sensitivity in the femtomole or low picomole range. In this study we have used high-accuracy, matrix-assisted laser desorption/ionization time of flight (MALDI TOF)-based peptide mapping to perform direct identification of distinct P450 isozymes in various rat and rabbit liver microsomes. For the first time, the P450 isozyme composition of clofibrate-induced rat and phenobarbital-induced rabbit liver microsomes was determined by peptide mass fingerprinting (PMF). Application of MALDI TOF-based PMF allows differential identification of such highly homologous P450s as CYP2B1 and CYP2B2. We have found that CYP2A10 previously reported only in rabbit olfactory and respiratory nasal mucosa is present in phenobarbital (PB)-induced rabbit liver microsomes. Two other rabbit P450s, earlier identified only by screening a cDNA library, were found to be present in PB-induced rabbit liver microsomes. In summary, direct identification of P450s by proteomic technique offers advantages over other methods with regard to identification of distinct P450 isozymes and should become a standard approach for characterizing microsomes. PMID- 12642459 TI - Homotropic versus heterotopic cooperativity of cytochrome P450eryF: a substrate oxidation and spectral titration study. AB - P450eryF is the only bacterial P450 to show cooperativity of substrate binding and oxidation. However, the studies reported so far have provided evidence only for homotropic cooperativity of P450eryF but not for heterotropic cooperativity. Therefore, oxidation of 7-benzyloxyquinoline (7-BQ) and 1-pyrenebutanol (1-PB) by P450eryF A245T and spectral binding of 9-aminophenanthrene (9-AP) to wild-type P450eryF were investigated in the presence of various effectors. The addition of steroids and flavones caused no stimulation but rather moderate inhibition of 7 BQ or 1-PB oxidation by P450eryF A245T. However, the binding affinity of 9-AP was significantly increased in the presence of androstenedione or alpha naphthoflavone (ANF). A comparative study with CYP3A4 revealed a similar increase in the binding affinity of 9-AP for the enzyme at low ANF concentrations but some competition at higher ANF concentrations. These studies, to our knowledge, provide the first report of heterotropic cooperativity in P450eryF as well as spectroscopic evidence for simultaneous presence of two ligand molecules in the CYP3A4 active site. PMID- 12642460 TI - The changing environment of graduate and postdoctoral training in drug metabolism: viewpoints from academia, industry, and government. AB - This article is an invited report of a symposium sponsored by the Drug Metabolism Division of the American Society for Pharmacology and Experimental Therapeutics held at Experimental Biology 2002 in New Orleans. The impetus for the symposium was a perceived shortage in the supply of graduate students qualified for drug metabolism research positions in industry, academia, and government. For industry, recent hiring stems largely from the expansion of drug metabolism departments in an effort to keep pace with the demands of drug discovery and new technologies. In turn, regulatory scientists are needed to review and verify the results of the increased number and volume of studies required for drug development and approval. Thus the initial source of training, academia, has been forced to recognize these external hiring pressures while trying to attract and retain the faculty, postdoctoral scientists, and students necessary for active teaching and research programs. The trend of the expansion of the interdisciplinary nature of traditional drug metabolism to include emerging technologies such as pharmacogenetics, transporters, and proteomics and the implications for future needs in training and funding were acknowledged. There was also consensus on the value of partnerships between academia and industry for increasing student interest and providing training in disciplines directly applicable to industrial drug metabolism research. Factors affecting the sources of these trainees, such as federal funding, the number of trainees per institution, and recent issues with immigration restrictions that have limited the flow of scientists were also discussed. PMID- 12642461 TI - Xanthine oxidase-catalyzed metabolism of 2-nitrofluorene, a carcinogenic air pollutant, in rat skin. AB - The reductive metabolism of 2-nitrofluorene, a carcinogenic air pollutant, in rat skin microsomes and cytosol was investigated. 2-Nitrofluorene was reduced to the corresponding amine by the microsomes with NADPH and by the cytosol with 2 hydroxypyrimidine or 4-hydroxypyrimidine under anaerobic conditions. The cytosolic activity was much higher than that of skin microsomes. The 2- or 4 hydroxypyrimidine-linked nitroreductase activity was inhibited by oxypurinol and (+/-)-8-(3-methoxy-4-phenylsulfinylphenyl) pyrazolo[1,5-a]-1,3,5-triazine-4(1H) one (BOF-4272), inhibitors of xanthine oxidase, but not by menadione, chlorpromazine and isovanillin, inhibitors of aldehyde oxidase. When skin cytosol was applied to a DEAE-cellulose column, the fractions containing xanthine oxidase exhibited a marked 2-hydroxypyrimidine-linked nitroreductase activity. In contrast, the aldehyde oxidase fraction showed little activity. Nitroreductase fractions obtained by ion exchange chromatography showed a band in Western blotting analysis using anti-rat xanthine oxidase. Moreover, the xanthine oxidase fraction exhibited a significant nitroreductase activity in the presence of 2 hydroxypyrimidine, 4-hydroxypyrimidine or hypoxanthine, and these activities were inhibited by inhibitors of xanthine oxidase. These results indicated that reduction of 2-nitrofluorene in the skin was mainly catalyzed by xanthine oxidase. PMID- 12642462 TI - Segmental intestinal transporters and metabolic enzymes on intestinal drug absorption. AB - Recently, a physiologically-based, segregated flow model that incorporates separate intestinal tissue and flow to both a nonabsorptive and an absorptive outermost layer (enterocytes) was shown to better describe the observations on route-dependent morphine glucuronidation in the rat small intestine than a traditional physiologically-based model. These theoretical models were expanded, as the segmental segregated flow model and the segmental traditional model, to view the intestine as three segments of equal lengths receiving equal flows to accommodate heterogeneities in segmental transporter and metabolic functions. The influence of heterogeneity in absorptive, exsorptive, and metabolic functions on drug clearance, bioavailability (F), and metabolite formation after intravenous and oral dosing was examined for the intestine when the tissue was the only organ of removal. Simulations were performed for first-order conditions, when drug partitioned readily (flow-limited distribution) or less readily (membrane-limited distribution) into intestinal tissue, and for different gastrointestinal transit times. The intestinal clearance was found to be inversely related to the rate constant for absorption of a drug that was subjected to secretion and was positively correlated with the metabolic and secretory intrinsic clearances. F was positively correlated with the absorption rate constant but was inversely related to the metabolic and secretory intrinsic clearances. The gastrointestinal transit time decreased metabolite formation, increased clearance, and decreased F. The simulations further showed that a descending metabolic intrinsic clearance yielded a lower F and an ascending segmental distribution of metabolic intrinsic clearance yielded a higher F. PMID- 12642463 TI - Inhibition and activation of the human liver microsomal and human cytochrome P450 3A4 metabolism of testosterone by deployment-related chemicals. AB - Cytochrome P450 (P450) enzymes are major catalysts involved in the metabolism of xenobiotics and endogenous substrates such as testosterone (TST). Major TST metabolites formed by human liver microsomes include 6beta-hydroxytestosterone (6beta-OHTST), 2beta-hydroxytestosterone (2beta-OHTST), and 15beta hydroxytestosterone (15beta-OHTST). A screen of 16 cDNA-expressed human P450 isoforms demonstrated that 94% of all TST metabolites are produced by members of the CYP3A subfamily with 6beta-OHTST accounting for 86% of all TST metabolites. Similar K(m) values were observed for production of 6beta-, 2beta-, and 15beta OHTST with human liver microsomes (HLM) and CYP3A4. However, V(max) and CL(int) were significantly higher for 6beta-OHTST than 2beta-OHTST (approximately 18 fold) and 15beta-OHTST (approximately 40-fold). Preincubation of HLM with a variety of ligands, including chemicals used in military deployments, resulted in varying levels of inhibition or activation of TST metabolism. The greatest inhibition of TST metabolism in HLM was following preincubation with organophosphorus compounds, including chlorpyrifos, phorate, and fonofos, with up to 80% inhibition noticed for several metabolites including 6beta-OHTST. Preincubation of CYP3A4 with chlorpyrifos, but not chlorpyrifos-oxon, resulted in 98% inhibition of TST metabolism. Phorate and fonofos also inhibited the production of most primary metabolites of CYP3A4. Kinetic analysis indicated that chlorpyrifos was one of the most potent inhibitors of major TST metabolites followed by fonofos and phorate. Chlorpyrifos, fonofos, and phorate inhibited major TST metabolites noncompetitively and irreversibly. Conversely, preincubation of CYP3A4 with pyridostigmine bromide increased metabolite levels of 6beta-OHTST and 2beta-OHTST. Preincubation of human aromatase (CYP19) with the test chemicals had no effect on the production of the endogenous estrogen, 17beta estradiol. PMID- 12642464 TI - Lipopolysaccharide-mediated modulation of cytochromes P450 in Stat1 null mice. AB - Signal transducer and activator of transcription (Stat), a family of transcriptional factors, has been demonstrated to play a critical role in gene regulation in response to inflammatory cytokines, such as interferon and interleukin-6. Inflammatory cytokines and bacterial endotoxin are known to suppress, in most of cases, the constitutive or induced cytochromes P450 (P450) in animals and humans. However, it is not clear if the suppression of P450 by cytokines is through the Stat-signaling pathway. In the present study, we determined whether Stat1 is involved in lipopolysaccharide (LPS)-mediated modulation of P450 in mouse liver. In both Stat1(+/+) (wild type) and Stat1(-/-) (null) mice, a single dose of LPS treatment (1 mg/kg of body weight, i.p.) significantly reduced the expression of CYP3A11, 2C29, and 1A2 mRNA to 8 to 40% of the control levels as determined by real-time quantitative reverse transcription-polymerase chain reaction. The reduction was supported by Western blot analysis. In contrast, LPS significantly induced the level of CYP4A10 mRNA in both Stat1(+/+) (338% of control) and Stat1(-/-) mice (264% of control). Although suppression of mRNA levels of CYP2E1, and 2D9 was not observed in either LPS-treated Stat1 null or wild-type animals, LPS treatment resulted in a reduction of CYP2E1 protein content, which was more significant in Stat1(+/+) (23% of control) than in Stat1(-/-) mice (67% of control). Consistent with this result, the chlorzoxazone 6-hydroxylase and lauric acid 11-hydroxylase activities, as CYP2E1 representative activities, were reduced markedly by LPS in Stat1(+/+) but not in Stat1(-/-) mice. The ethoxyresorufin O-deethylase activity, as a representative CYP1A activity, was also reduced significantly only in LPS treated Stat1(+/+) mice. These data clearly demonstrate that LPS-mediated modulation of CYP3A11, 2B10, 2C29, 1A2, and 4A10 in mouse liver is Stat1 independent. However, the significant difference between the LPS-treated Stat1(+/+) and Stat1(-/-) mice in the levels of CYP2E1 protein and activity as well as in the activity level of CYP1A suggests that Stat1 may be indirectly involved in the post-transcriptional modulation of these two mouse P450 enzymes. PMID- 12642465 TI - Functional characterization of cytochrome P450 2B6 allelic variants. AB - Cytochrome P450 (P450) 2B6 is a hepatic enzyme of potential importance for the metabolism of clinically used drugs and environmental or abused toxicants. Genetic polymorphisms of CYP2B6 (CYP2B6*2, CYP2B6*3, CYP2B6*4, CYP2B6*5, CYP2B6*6 and CYP2B6*7; wild-type, CYP2B6*1) were found previously in white and Japanese populations. In the present study, the goal was to investigate the effects of amino acid substitutions on CYP2B6 function. Wild-type (CYP2B6.1) and all of the known variants of CYP2B6 (CYP2B6.2, CYP2B6.3, CYP2B6.4, CYP2B6.5, CYP2B6.6, and CYP2B6.7) were transiently expressed in COS-1 cells, and their 7-ethoxy-4 trifluoromethylcoumarin O-deethylation activities were determined. The levels of the variant CYP2B6 proteins were relatively low compared with that of CYP2B6.1, although the differences were not significant. The activities of 7-ethoxy-4 trifluoromethylcoumarin O-deethylation on the basis of the CYP2B6 protein level at low (0.5 microM) and high (50 microM) substrate concentrations varied among wild-type and variant CYP2B6 proteins. All CYP2B6 enzymes showed typical Michaelis-Menten kinetics. The K(m) value of CYP2B6.6 was significantly higher than that of CYP2B6.1. Those CYP2B6 variants having a Lys262Arg substitution (CYP2B6.4, CYP2B6.6, and CYP2B6.7) showed increased values for V(max) and V(max)/K(m), whereas the kinetic parameters of CYP2B6.2 and CYP2B6.3 were not affected by the corresponding amino acid substitution. These results may mean that Lys262 in combination with other amino acid residues such as Gln172 and Arg487 is associated with the CYP2B6 function and that the genetic polymorphism of CYP2B6 leads to interindividual differences in xenobiotic metabolism. PMID- 12642466 TI - Identification of the human liver enzymes involved in the metabolism of the antimigraine agent almotriptan. AB - Almotriptan is a novel highly selective 5-hydroxytryptamine(1B/1D) agonist developed for the acute oral treatment of migraine. The in vitro metabolism of almotriptan has been investigated using human liver subcellular fractions and cDNA-expressed human enzymes, to study the metabolic pathways and identify the enzymes responsible for the formation of the major metabolites. Specific enzymes were identified by correlation analysis, chemical inhibition studies, and incubation with various cDNA expressed human enzymes. Human liver microsomes and S9 fraction metabolize almotriptan by 2-hydroxylation of the pyrrolidine group to form a carbinolamine metabolite intermediate, a reaction catalyzed by CYP3A4 and CYP2D6. This metabolite is further oxidized by aldehyde dehydrogenase to the open ring gamma-aminobutyric acid metabolite. Almotriptan is also metabolized at the dimethylaminoethyl group by N-demethylation, a reaction that is carried out by five different cytochrome P450s, flavin monooxygenase-3 mediated N-oxidation, and MAO-A catalyzed oxidative deamination to form the indole acetic acid and the indole ethyl alcohol derivatives of almotriptan. The use of human liver mitochondria confirmed the contribution of MAO-A to the metabolism of almotriptan. Both, the gamma-aminobutyric acid and the indole acetic acid metabolites have been found to be the major in vivo metabolites of almotriptan in humans. In addition, different clinical trials conducted to study the effects of CYP3A4, CYP2D6, and MAO-A on the pharmacokinetics of almotriptan confirmed the involvement of these enzymes in the metabolic clearance of this drug and that no dose changes are required in the presence of inhibitors of these enzymes. PMID- 12642468 TI - Effects of prototypical microsomal enzyme inducers on cytochrome P450 expression in cultured human hepatocytes. AB - Cultured human hepatocytes are a valuable in vitro system for evaluating new molecular entities as inducers of cytochrome P450 (P450) enzymes. The present study summarizes data obtained from 62 preparations of cultured human hepatocytes that were treated with vehicles (saline or dimethylsulfoxide, 0.1%), beta naphthoflavone (33 microM), phenobarbital (100 or 250 microM), isoniazid (100 microM) and/or rifampin (20 or 50 microM), and examined for the expression of P450 enzymes based on microsomal activity toward marker substrates, or in the case of CYP2C8, the level of immunoreactive protein. The results show that CYP1A2 activity was markedly induced by beta-naphthoflavone (on average 13-fold, n = 28 preparations), and weakly induced by phenobarbital (1.9-fold, n = 25) and rifampin (2.3-fold, n = 22); CYP2A6 activity tended to be increased with phenobarbital (n = 7) and rifampin (n = 3) treatments, but the effects were not statistically significant; CYP2B6 was induced by phenobarbital (6.5-fold, n = 13) and rifampin (13-fold, n = 14); CYP2C8 was induced by phenobarbital (4.0-fold, n = 4) and rifampin (5.2-fold, n = 4); CYP2C9 was induced by phenobarbital (1.8 fold, n = 14) and rifampin (3.5-fold, n = 10); CYP2C19 was markedly induced by rifampin (37-fold, n = 10), but relatively modestly by phenobarbital (7-fold, n = 9); CYP2D6 was not significantly induced by phenobarbital (n = 5) or rifampin (n = 5); CYP2E1 was induced by phenobarbital (1.7-fold, n = 5), rifampin (2.2-fold, n = 5), and isoniazid (2.3-fold, n = 5); and, CYP3A4 was induced by phenobarbital (3.3-fold, n = 42) and rifampin (10-fold, n = 61), but not by beta naphthoflavone. Based on these observations, we generalize that beta naphthoflavone induces CYP1A2 and isoniazid induces CYP2E1, whereas rifampin and, to a lesser extent phenobarbital, tend to significantly and consistently induce enzymes of the CYP2A, CYP2B, CYP2C, CYP2E, and CYP3A subfamilies but not the 2D subfamily. PMID- 12642467 TI - Characterization of substrate binding to cytochrome P450 1A1 using molecular modeling and kinetic analyses: case of residue 382. AB - Key residue Val-382 in P450 1A1 has been predicted to interact with the alkoxy chain of resorufin derivatives. Therefore, we undertook a detailed analysis of substrate mobility in the active site of the P450 1A1 homology model and assessed the effect of mutations at position 382. Dynamic trajectories of 7-methoxy-, 7 ethoxy-, and 7-pentoxyresorufin indicated that 7-ethoxyresorufin would be oxidized most efficiently by the wild-type enzyme. The Val-382-->Ala mutation would increase the O-dealkylation of 7-pentoxyresorufin but decrease the oxidation of other substrates. In the case of the V382L mutant, the large bulk of Leu would block alkoxyresorufins from productive binding orientations leading to lowered activities. Binding free energy calculations for three substrates with 1A1 WT and two mutants indicated that binding constants would be similar for all enzyme-substrate combinations. Modeling predictions were tested experimentally. The plasmid containing the cDNA for human P450 1A1 modified for bacterial expression was altered to include a C-terminal PCR-generated six histidine domain to facilitate enzyme purification. The V382A and V382L mutants were constructed by site-directed mutagenesis and Escherichia coli-expressed enzymes purified using Ni-NTA affinity chromatography. The activity of the WT 1A1 was highest toward 7-ethoxyresorufin and lowest toward 7-pentoxyresorufin. Both mutants displayed a decrease in V(max) with 7-methoxy- and 7-ethoxyresorufin, whereas for the V382A mutant, V(max) with 7-pentoxyresorufin was increased. No significant changes in K(m) were observed relative to the wild-type enzyme. The experimental results are thus in good agreement with modeling predictions. PMID- 12642469 TI - CYP3A4 and CYP3A7-mediated carbamazepine 10,11-epoxidation are activated by differential endogenous steroids. AB - Recently, we reported that several endogenous steroids affect CYP3A4-mediated drug metabolism, using human adult liver microsomes as an enzyme source. Especially, carbamazepine (CBZ) 10,11-epoxidation is activated by androstenedione (AND). In the present studies, we investigated the effects of endogenous steroids on the activity of CBZ 10,11-epoxidation by expressed CYP3A4 and CYP3A7. When expressed CYP3A4 was used as an enzyme source, the addition of AND to the reaction mixture also caused a drastic increase in the activity of CBZ 10,11 epoxidase, and resulted in a change in the kinetics from sigmoid to Michaelis Menten type. On the other hand, expressed CYP3A7-mediated CBZ 10,11-epoxidation was activated by sulfate conjugate steroids, such as pregnenolone 3-sulfate, 17alpha-hydroxypregnenolone 3-sulfate, and dehydroepiandrosterone 3-sulfate (DHEA S), whereas the unconjugated form corresponding to these three steroids did not activate the reaction. Especially, DHEA-S was found to be a potent activator of CBZ 10,11-epoxidation by expressed CYP3A7. The kinetic character of CBZ 10,11 epoxidation by CYP3A7 is Michaelis-Menten type regardless of the presence of DHEA S. The presence of DHEA-S caused a decrease in K(m) and increase in V(max) for CYP3A7-mediated CBZ 10,11-epoxidation, whereas DHEA-S 16alpha-hydroxylation was not affected by the coexistence of CBZ. In conclusion, CYP3A4 and CYP3A7-mediated CBZ 10,11-epoxidations are activated by different types of endogenous steroids. This is the first report regarding CYP3A7 cooperativity. PMID- 12642470 TI - Comparative effects of thiazolidinediones on in vitro P450 enzyme induction and inhibition. AB - Rosiglitazone and pioglitazone are thiazolidinediones used for treatment of noninsulin-dependent diabetes mellitus. These compounds, along with troglitazone, were evaluated for the ability to induce cytochrome P450 enzymes (P450) in primary human hepatocyte cultures and to inhibit P450 in human microsomes. In induction studies, all three thiazolidinediones caused a dose-dependent increase in CYP3A4 activity and immunoreactive protein. While troglitazone was the most potent, rosiglitazone and pioglitazone generally exceeded troglitazone in absolute CYP3A4 activity achieved at concentrations > or =10 microM. A comparable concentration-dependent increase in CYP2B6 immunoreactive protein was observed with all three thiazolidinediones. Microarray analysis revealed rifampin > troglitazone > pioglitazone > rosiglitazone in terms of CYP3A4 mRNA induction potential with 10 microM compound. Inhibition studies conducted for CYP1A2, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A4, CYP2A6, and CYP2E1 showed troglitazone to be the most nonselective and potent inhibitor followed by rosiglitazone and pioglitazone. In vitro, the thiazolidinediones were strong inhibitors of CYP2C8, with K(i) values between 1.7 and 5.6 microM, and of CYP3A4, with K(i) values between 1.6 and 11.8 microM. Troglitazone, in addition, inhibited CYP2C9 (K(i) 0.6 microM). Although the inhibitory effects of the thiazolidinediones have not been demonstrated clinically, our results suggest there is potential for interactions with CYP2C8 substrates. This is the first report of in vitro induction of P450 enzymes by rosiglitazone and pioglitazone. While only the induction of CYP3A4 by troglitazone has been demonstrated in vivo, these results suggest that other thiazolidinediones may have the potential to cause clinically significant drug interactions at sufficiently high doses. PMID- 12642471 TI - Retention of transporter activities in cryopreserved, isolated rat hepatocytes. AB - The success of cryopreservation of isolated hepatocytes with existing methodologies is assessed with respect to the retentivity of cell integrity/viability (defined by trypan blue) and metabolic activities upon thawing in comparison to those of freshly prepared cells. But the ability of the cryopreserved cells to transport xenobiotics relative to that of freshly prepared cells has not been investigated. In this study, we optimized our previous methodology for cryopreservation and evaluated the metabolism and transport of thawed hepatocytes. Half of the freshly, isolated rat hepatocytes prepared by collagenase perfusion were immediately used for studies of transport of [(14)C]taurocholate, [(3)H]estrone sulfate and [(3)H]estradiol 17beta-D glucuronide (1 microM) and metabolism of 7-hydroxy-4-(trifluoromethyl)-coumarin (100 microM), (3,4-difluorobenzyloxy)-5,5-dimethyl-4-(4-methylsulfonylphenyl) (5H)-furan-2-one (250 microM), bufuralol (100 microM), and tolbutamide (100 microM), probes for UDP-glucuronyl transferase (UGT) and CYP3A, CYP2D, and CYP2C, respectively. The remaining half was cryopreserved using an optimized, programmed freezing protocol, which was developed to minimize the prolonged release of latent heat during freezing. With the exception of the UGT probe, no significant difference (P > 0.05) was found in both metabolism and transport with freshly isolated versus cryopreserved hepatocytes upon thawing. In conclusion, we have demonstrated for the first time that thawed rat hepatocytes cryopreserved by a programmed-freezing protocol retain drug transport activities. PMID- 12642472 TI - Glucuronides of tea catechins: enzymology of biosynthesis and biological activities. AB - (-)-Epigallocatechin gallate (EGCG) and (-)-epigallocatechin (EGC) are major green tea catechins with antioxidant and anticancer activities. In this study, we characterized the glucuronidation of EGCG and EGC in human, mouse, and rat microsomes and by nine different human UGT 1A and 2B isozymes expressed in insect cells. Six EGCG and EGC glucuronides were biosynthesized, and their structures were identified for the first time. (-)-EGCG-4"-O-glucuronide was the major EGCG glucuronide formed in all incubations. The catalytic efficiency (V(max)/K(m)) for (-)-EGCG-4"-O-glucuronide formation followed the order: mouse intestine > mouse liver > human liver > rat liver >> rat small intestine. The UGT-catalyzed glucuronidation of EGC was much lower than that of EGCG. The V(max)/K(m) for (-) EGC-3'-O-glucuronide followed the following order: mouse liver > human liver > rat liver > rat and mouse small intestine. Human UGT1A1, 1A8, and 1A9 had high activities with EGCG. UGT1A8, an intestine-specific UGT, had the highest V(max)/K(m) for EGCG but low activity with EGC. Mice appeared to be more similar to humans than rats to humans in the glucuronidation of EGCG and EGC. Some of these catechin glucuronides retained the activities of their parent compounds in radical scavenging and in inhibiting the release of arachidonic acid from HT-29 human colon cancer cells. These results provide foundations for understanding the biotransformation and biological activities of tea catechins. PMID- 12642473 TI - Modeling the metabolism of idarubicin to idarubicinol in rat heart: effect of rutin and phenobarbital. AB - Since the severe cardiotoxicity of anthracyclines has been attributed to the intramyocardial formation of C-13 alcohol metabolites, the kinetics of cardiac metabolite formation and disposition as well as the effect of carbonyl reductase inhibitors are of specific interest. This study was designed to investigate the effect of rutin and phenobarbital on the pharmacokinetics of idarubicin (IDA) and its conversion to idarubicinol (IDOL) in the single-pass perfused rat heart. After infusion of IDA (0.5 mg) during 1min, the venous outflow concentrations of IDA and IDOL were measured up to 80 min in the presence and absence of rutin and phenobarbital. A kinetic model was developed to help to interpret the concentration profiles in terms of compartmentation of IDOL formation and to estimate parameters quantitatively descriptive of the transport and biotransformation processes. Rutin and phenobarbital significantly reduced the residual amount of IDOL in heart to 64 and 47% of control, respectively. Pharmacokinetic modeling of the data revealed that IDOL is generated in two different compartments, besides the tissue compartment characterized by saturable uptake, also the compartment that accounts for the quasi-instantaneous initial distribution process is involved. The efflux rate constant of IDOL, k(21,IDOL,) was much smaller than that of IDA. Rutin and phenobarbital significantly reduced IDOL production. Additionally, phenobarbital competitively inhibited the saturable uptake of both IDA and IDOL (increase in apparent Michaelis constants). Reanalysis of data obtained in previous experiments showed that P-glycoprotein inhibitors (verapamil and amiodarone) reduced IDOL uptake in a similar way as already shown for IDA. The present study further supports the utility of pharmacokinetic modeling in identifying sites of drug interactions within the heart. PMID- 12642474 TI - Thalidomide-induced suppression of embryo fibroblast proliferation requires CYP1A1-mediated activation. AB - An enzyme involved in the metabolic activation of thalidomide has been investigated using embryo fibroblast proliferation as a marker. Thalidomide (30 microM) induced-suppression of embryo fibroblast proliferation was detected in the presence of liver microsomes from rabbit but not from mouse. The addition of a selective inhibitor of CYP1A, alpha-naphthoflavone (4 microM), or furafylline (4 microM), to the incubation mixture abolished the thalidomide-induced suppression. Furthermore, addition of anti-rat CYP1A1 antibody also resulted in inhibition of suppression. The thalidomide-induced suppression was also observed with the microsomal system from human HepG2 cells pretreated with 3 methylcholanthrene (10 microM) but not from those pretreated with the vehicle. Both CYP1A1 and CYP1A2 proteins were detected in the rabbit liver microsomes by immunoblot analyses, but only CYP1A2 protein was detected in the mouse liver microsomes. In addition, CYP1A1 protein was detected in microsomes from HepG2 cells pretreated with 3-methylcholanthrene but not with the vehicle. These results strongly suggest the involvement of CYP1A1 in the thalidomide-induced suppression of embryo fibroblast proliferation. PMID- 12642475 TI - Cytochrome P450 2C8 and flavin-containing monooxygenases are involved in the metabolism of tazarotenic acid in humans. AB - Upon oral administration, tazarotene is rapidly converted to tazarotenic acid by esterases. The main circulating agent, tazarotenic acid is subsequently oxidized to the inactive sulfoxide metabolite. Therefore, alterations in the metabolic clearance of tazarotenic acid may have significant effects on its systemic exposure. The objective of this study was to identify the human liver microsomal enzymes responsible for the in vitro metabolism of tazarotenic acid. Tazarotenic acid was incubated with 1 mg/ml pooled human liver microsomes, in 100 mM potassium phosphate buffer (pH 7.4), at 37 degrees C, over a period of 30 min. The microsomal enzymes that may be involved in tazarotenic acid metabolism were identified through incubation with microsomes containing cDNA-expressed human microsomal isozymes. Chemical inhibition studies were then conducted to confirm the identity of the enzymes potentially involved in tazarotenic acid metabolism. Reversed-phase high performance liquid chromatography was used to quantify the sulfoxide metabolite, the major metabolite of tazarotenic acid. Upon incubation of tazarotenic acid with microsomes expressing CYP2C8, flavin-containing monooxygenase 1 (FMO1), or FMO3, marked formation of the sulfoxide metabolite was observed. The involvement of these isozymes in tazarotenic acid metabolism was further confirmed by inhibition of metabolite formation in pooled human liver microsomes by specific inhibitors of CYP2C8 or FMO. In conclusion, the in vitro metabolism of tazarotenic acid to its sulfoxide metabolite in human liver microsomes is mediated by CYP2C8 and FMO. PMID- 12642476 TI - Multiple alterations of canalicular membrane transport activities in rats with CCl(4)-induced hepatic injury. AB - The influence of CCl(4)-induced experimental hepatic injury (CCl(4)-EHI) on the expression and transport activities of primary active transporters on the canalicular membrane, including P-glycoprotein (P-gp), a bile salt export pump (Bsep) and a multidrug resistance associated protein2 (Mrp2), was assessed. CCl(4)-EHI was induced by an intraperitoneal injection of CCl(4) to rats at a dose of 1 ml/kg 24 h prior to the preparation of canalicular liver plasma membrane (cLPM) vesicles and pharmacokinetic studies. The expression of each transporter was measured for the vesicles via Western blot analysis at 6, 12, 24, 36, and 48 h after the injection of CCl(4). The in vivo canalicular excretion clearance (CL(exc)) of [(3)H]daunomycin, [(3)H]taurocholate and [(3)H]17beta estradiol-17beta-D-glucuronide (E(2)17betaG), representative substrates of P-gp, Bsep, and Mrp2, respectively, was determined following an i.v. infusion to rats. The uptake of each substrate into cLPM vesicles in the presence of ATP was also measured by a rapid filtration technique. As the result of the CCl(4)-EHI, the protein level of transporters was altered as a function of time in multiple manners; it was increased by 3.6-fold for P-gp, unchanged for Bsep, and decreased by 73% for Mrp2 at 24 h. The in vivo CL(exc) and the intrinsic uptake clearance into cLPM vesicles (CL(int)) at 24 h after the CCl(4) injection (CCl(4)-EHI(24 h)) were also influenced by the EHI in a similar manner; they were increased by 1.8- and 1.9-fold for daunomycin, unchanged for taurocholate, and decreased by 41 and 39% for E(2)17betaG, respectively, consistent with multiple alterations in the expression of the relevant transporters. PMID- 12642477 TI - Pharmacokinetics and metabolism of a COX-2 inhibitor, valdecoxib, in mice. AB - The pharmacokinetics and metabolism of valdecoxib, a potent cyclooxygenase-2 selective inhibitor, were investigated in mice. Valdecoxib was extensively metabolized after a single 5 mg/kg oral administration of [(14)C]valdecoxib and elimination of unchanged drug was minor (less than 1%) in male and female mice. The total mean percentage of administered radioactive dose recovered was 99.8% in the male mice and 94.7% in the female mice. Sixteen metabolites were identified in mouse plasma, red blood cells, urine, and feces. The main phase I metabolic pathway of valdecoxib in mice involved the oxidation of the 5-methyl group to form the active hydroxymethyl metabolite M1. M1 was further oxidized to the carboxylic acid metabolite M4, which underwent opening of the isoxazole ring to form M6 and M13. Phase II metabolism included glucuronide, glucoside, and methyl sulfone conjugations. M1 was also conjugated with glucuronic acid and glucose to yield M-G and M1-glucose, respectively. Three novel methylsulfone conjugates M20, M21, and M21-G were detected in blood or urine. Valdecoxib and M1 were the major radioactive components in plasma and red blood cells. The plasma area under the curve from zero to infinity (AUC(0-infinity)) values for valdecoxib and M1 were 3.58 and 0.850 microg. h/ml in males and 2.08 and 1.63 microg. h/ml in females, respectively. The RBC AUC(0-infinity) values for valdecoxib and M1 were 12.1 and 22.6 microg. h/g in males and 6.42 and 35.2 microg. h/g in females, respectively. PMID- 12642478 TI - Pharmacokinetics, metabolic stability, and subcutaneous bioavailability of a genetically engineered analog of DcR3, FLINT [DcR3(R218Q)], in cynomolgus monkeys and mice. AB - Decoy receptor 3 (DcR3) is a novel member of the tumor necrosis factor receptor superfamily, which binds to and blocks the activities of the ligands, FasL and LIGHT (a cellular ligand for herpes virus entry mediator and lymphotoxin receptor), that play an important role in regulating apoptosis in normal physiology. DcR3 was rapidly degraded to a major circulating metabolic fragment, DcR3(1-218), after subcutaneous administration in primates and mice. DcR3 was molecularly engineered by changing the arginine residue at position 218 to glutamine to generate a potentially stable analog, DcR3(R218Q), which we termed FasLigand inhibitor protein [FLINT (LY498919)]. The influence of this modification on the kinetics and bioavailability of DcR3 was evaluated in primates and mice. After i.v. administration of FLINT and DcR3, both compounds were cleared from the plasma in a bi-phasic manner, with the terminal phase half life being somewhat longer for FLINT than for DcR3. After s.c. administration, the exposure to the full-length form of FLINT was 5.7- to 6-fold greater than for DcR3. In both primates and mice, greater than 90% of circulating immunoreactivity after s.c. administration of FLINT was associated with intact molecule, whereas only 17 to 37% was associated with intact molecule after administration of DcR3. The absolute s.c. bioavailability of intact FLINT was approximately 4- to 6-fold higher than for DcR3. The improved s.c. bioavailability of FLINT is related to the increased metabolic stability afforded to the molecule as a result of the amino acid mutation at position 218 of the primary sequence of DcR3 and may translate to the need for lower therapeutic doses in a number of disease indications. PMID- 12642479 TI - The left-right determinant Inversin is a component of node monocilia and other 9+0 cilia. AB - Inversin (Inv), a protein that contains ankyrin repeats, plays a key role in left right determination during mammalian embryonic development, but its precise function remains unknown. Transgenic mice expressing an Inv and green fluorescent protein (GFP) fusion construct (Inv::GFP) were established to facilitate characterization of the subcellular localization of Inv. The Inv::GFP transgene rescued the laterality defects and polycystic kidney disease of Inv/Inv mice, indicating that the fusion protein is functional. In transgenic embryos, Inv::GFP protein was detected in the node monocilia. The fusion protein was also present in other 9+0 monocilia, including those of kidney epithelial cells and the pituitary gland, but it was not localized to 9+2 cilia. The N-terminal region of Inv (InvDeltaC) including the ankyrin repeats also localized to the node cilia and rescued the left-right defects of Inv/Inv mutants. Although no obvious abnormalities were detected in the node monocilia of Inv/Inv embryos, the laterality defects of such embryos were corrected by an artificial leftward flow of fluid in the node, suggesting that nodal flow is impaired by the Inv mutation. These results suggest that the Inv protein contributes to left-right determination as a component of monocilia in the node and is essential for the generation of normal nodal flow. PMID- 12642480 TI - Early patterning of the spider embryo: a cluster of mesenchymal cells at the cumulus produces Dpp signals received by germ disc epithelial cells. AB - In early embryogenesis of spiders, the cumulus is characteristically observed as a cellular thickening that arises from the center of the germ disc and moves centrifugally. This cumulus movement breaks the radial symmetry of the germ disc morphology, correlating with the development of the dorsal region of the embryo. Classical experiments on spider embryos have shown that a cumulus has the capacity to induce a secondary axis when transplanted ectopically. In this study, we have examined the house spider, Achaearanea tepidariorum, on the basis of knowledge from Drosophila to characterize the cumulus at the cellular and molecular level. In the cumulus, a cluster of about 10 mesenchymal cells, designated the cumulus mesenchymal (CM) cells, is situated beneath the epithelium, where the CM cells migrate to the rim of the germ disc. Germ disc epithelial cells near the migrating CM cells extend cytoneme-like projections from their basal side onto the surface of the CM cells. Molecular cloning and whole-mount in situ hybridization showed that the CM cells expressed a spider homolog of Drosophila decapentaplegic (dpp), which encodes a secreted protein that functions as a dorsal morphogen in the Drosophila embryo. Furthermore, the spider Dpp signal appeared to induce graded levels of the phosphorylated Mothers against dpp (Mad) protein in the nuclei of germ disc epithelial cells. Adding data from spider homologs of fork head, orthodenticle and caudal, we suggest that, in contrast to the Drosophila embryo, the progressive mesenchymal epithelial cell interactions involving the Dpp-Mad signaling cascade generate dorsoventral polarity in accordance with the anteroposterior axis formation in the spider embryo. Our findings support the idea that the cumulus plays a central role in the axial pattern formation of the spider embryo. PMID- 12642481 TI - A zone of frontonasal ectoderm regulates patterning and growth in the face. AB - A fundamental set of patterning genes may define the global organization of the craniofacial region. One of our goals has been to identify these basic patterning genes and understand how they regulate outgrowth of the frontonasal process, which gives rise to the mid and upper face. We identified a molecular boundary in the frontonasal process ectoderm, defined by the juxtaposed domains of Fibroblast growth factor 8 and Sonic hedgehog, which presaged the initial site of frontonasal process outgrowth. Fate maps confirmed that this boundary region later demarcated the dorsoventral axis of the upper beak. Ectopic transplantation of the ectodermal boundary region activated a cascade of molecular events that reprogrammed the developmental fate of neural crest-derived mesenchyme, which resulted in duplications of upper and lower beak structures. We discuss these data in the context of boundary/morphogen models of patterning, and in view of the recent controversy regarding neural crest pre-patterning versus neural crest plasticity. PMID- 12642482 TI - Cell-autonomous involvement of Mab21l1 is essential for lens placode development. AB - The mab-21 gene was first identified because of its requirement for ray identity specification in Caenorhabditis elegans. It is now known to constitute a family of genes that are highly conserved from vertebrates to invertebrates, and two homologs, Mab21l1 and Mab21l2, have been identified in many species. We describe the generation of Mab21l1-deficient mice with defects in eye and preputial gland formation. The mutant mouse eye has a rudimentary lens resulting from insufficient invagination of the lens placode caused by deficient proliferation. Chimera analyses suggest that the lens placode is affected in a cell-autonomous manner, although Mab21l1 is expressed in both the lens placode and the optic vesicle. The defects in lens placode development correlate with delayed and insufficient expression of Foxe3, which is also required for lens development, while Maf, Sox2, Six3 and PAX6 levels are not significantly affected. Significant reduction of Mab21l1 expression in the optic vesicle and overlying surface ectoderm in Sey homozygotes indicates that Mab21l1 expression in the developing eye is dependent upon the functions of Pax6 gene products. We conclude that Mab21l1 expression dependent on PAX6 is essential for lens placode growth and for formation of the lens vesicle; lack of Mab21l1 expression causes reduced expression of Foxe3 in a cell-autonomous manner. PMID- 12642483 TI - The bHLH protein Dimmed controls neuroendocrine cell differentiation in Drosophila. AB - Neuroendocrine cells are specialized to produce, maintain and release large stores of secretory peptides. We show that the Drosophila dimmed/Mist1 bHLH gene confers such a pro-secretory phenotype on neuroendocrine cells. dimmed is expressed selectively in central and peripheral neuroendocrine cells. In dimmed mutants, these cells survive, and adopt normal cell fates and morphology. However, they display greatly diminished levels of secretory peptide mRNAs, and of diverse peptides and proteins destined for regulated secretion. Secretory peptide levels are lowered even in the presence of artificially high secretory peptide mRNA levels. In addition, overexpression of dimmed in a wild-type background produces a complimentary phenotype: an increase in secretory peptide levels by neuroendocrine cells, and an increase in the number of cells displaying a neuroendocrine phenotype. We propose that dimmed encodes an integral component of a novel mechanism by which diverse neuroendocrine lineages differentiate and maintain the pro-secretory state. PMID- 12642484 TI - XMAN1, an inner nuclear membrane protein, antagonizes BMP signaling by interacting with Smad1 in Xenopus embryos. AB - A family of inner nuclear membrane proteins is implicated in gene regulation by interacting with chromatin, nuclear lamina and intranuclear proteins; however, the physiological functions of these proteins are largely unknown. Using a Xenopus expression screening approach with an anterior neuroectoderm cDNA library, we have identified an inner nuclear membrane protein, XMAN1, as a novel neuralizing factor that is encoded by the Xenopus ortholog of human MAN1. XMAN1 mRNA is expressed maternally, and appears to be restricted to the entire ectoderm at the early gastrula stage, then to the anterior neuroectoderm at the neurula stage. XMAN1 induces anterior neural markers without mesoderm induction in ectodermal explants, and a partial secondary axis when expressed ventrally by dorsalizing the ventral mesoderm. Importantly, XMAN1 antagonizes bone morphogenetic protein (BMP) signaling downstream of its receptor Alk3, as judged by animal cap assays, in which XMAN1 blocks expression of downstream targets of BMP signaling (Xhox3 and Msx1), and by luciferase reporter assays, in which XMAN1 suppresses BMP-dependent activation of the Xvent2 promoter. Deletion mutant analyses reveal that the neuralizing and BMP-antagonizing activities of XMAN1 reside in the C-terminal region, and that the C-terminal region binds to Smad1, Smad5 and Smad8, which are intracellular mediators of the BMP pathway. Interference with endogenous XMAN1 functions with antisense morpholino oligos leads to the reduction of anterior neuroectoderm. These results provide the first evidence that the nuclear envelope protein XMAN1 acts as a Smad-interacting protein to antagonize BMP signaling during Xenopus embryogenesis. PMID- 12642485 TI - Nodal signaling induces the midline barrier by activating Nodal expression in the lateral plate. AB - The transcription factor Foxh1 mediates Nodal signaling. The role of Foxh1 in left-right (LR) patterning was examined with mutant mice that lack this protein in lateral plate mesoderm (LPM). The mutant mice failed to express Nodal, Lefty2 and Pitx2 on the left side during embryogenesis and exhibited right isomerism. Ectopic introduction of Nodal into right LPM, by transplantation of left LPM or by electroporation of a Nodal vector, induced Nodal expression in wild-type embryos but not in the mutant. Ectopic Nodal expression in right LPM also induced Lefty1 expression in the floor plate. Nodal signaling thus initiates asymmetric Nodal expression in LPM and induces Lefty1 at the midline. Monitoring of Nodal activity in wild-type and Foxh1 mutant embryos suggested that Nodal activity travels from the node to left LPM, and from left LPM to the midline. PMID- 12642486 TI - A role for actin dynamics in individualization during spermatogenesis in Drosophila melanogaster. AB - In order to better understand the mechanism of sperm individualization during spermatogenesis in Drosophila melanogaster, we have developed an in vitro culture system in which we can perform live observation of individualization in isolated cysts. The whole process of individualization, during which a bundle of 64 syncytial spermatids is separated into individual sperm, takes place in these cultures. Individualization complexes, which consist of 64 cones of actin that assemble around the sperm nuclei, move to the basal end of the tails, forming a characteristic "cystic bulge" that contains an accumulation of cytoplasm, syncytial membrane and vesicles. The cystic bulge is the site of membrane remodeling and its movement was used to follow the progress of individualization. The speed of cystic bulge movement is fairly constant along the length of the cyst. Actin drugs, but not microtubule drugs inhibit cystic bulge movement, suggesting that the movement requires proper actin dynamics but not microtubules. GFP-tagged actin was expressed in the cyst and fluorescence recovery after photobleaching was monitored using confocal microscopy to analyze actin dynamics in cones. Actin turns over throughout the cone, with that at the leading edge of the cones turning over with slightly faster kinetics. Actin does not treadmill from the front to the back of the cone. Actin in moving actin cones turns over in about 12 minutes, although prior to onset of movement, turnover is much slower. Visualization of membrane using FM1-43 reveals that the cystic bulge has an extremely complicated series of membrane invaginations and the transition from syncytial to individualized spermatids occurs at the front of the actin cones. We also suggest that endocytosis and exocytosis might not be important for membrane remodeling. This system should be suitable for analysis of defects in male sterile mutants and for investigating other steps of spermatogenesis. PMID- 12642487 TI - Effects of tethering HP1 to euchromatic regions of the Drosophila genome. AB - Heterochromatin protein 1 (HP1) is a conserved non-histone chromosomal protein enriched in heterochromatin. On Drosophila polytene chromosomes, HP1 localizes to centric and telomeric regions, along the fourth chromosome, and to specific sites within euchromatin. HP1 associates with centric regions through an interaction with methylated lysine nine of histone H3, a modification generated by the histone methyltransferase SU(VAR)3-9. This association correlates with a closed chromatin configuration and silencing of euchromatic genes positioned near heterochromatin. To determine whether HP1 is sufficient to nucleate the formation of silent chromatin at non-centric locations, HP1 was tethered to sites within euchromatic regions of Drosophila chromosomes. At 25 out of 26 sites tested, tethered HP1 caused silencing of a nearby reporter gene. The site that did not support silencing was upstream of an active gene, suggesting that the local chromatin environment did not support the formation of silent chromatin. Silencing correlated with the formation of ectopic fibers between the site of tethered HP1 and other chromosomal sites, some containing HP1. The ability of HP1 to bring distant chromosomal sites into proximity with each other suggests a mechanism for chromatin packaging. Silencing was not dependent on SU(VAR)3-9 dosage, suggesting a bypass of the requirement for histone methylation. PMID- 12642488 TI - RhoA and Rac1 GTPases mediate the dynamic rearrangement of actin in peripheral glia. AB - Peripheral glial cells in both vertebrates and insects are born centrally and travel large distances to ensheathe axons in the periphery. There is very little known about how this migration is carried out. In other cells, it is known that rearrangement of the Actin cytoskeleton is an integral part of cell motility, yet the distribution of Actin in peripheral glial cell migration in vivo has not been previously characterized. To gain an understanding of how glia migrate, we specifically labeled the peripheral glia of Drosophila melanogaster using an Actin-GFP marker and analyzed their development in the embryonic PNS. It was found that Actin cytoskeleton is dynamically rearranged during glial cell migration. The peripheral glia were observed to migrate as a continuous chain of cells, with the leading glial cells appearing to participate to the greatest extent in exploring the extracellular surroundings with filopodia-like Actin containing projections. We hypothesized that the small GTPases Rho, Rac and Cdc42 are involved in Actin cytoskeletal rearrangements that underlie peripheral glial migration and nerve ensheathement. To test this, transgenic forms of the GTPases were ectopically expressed specifically in the peripheral glia during their migration and wrapping phases. The effects on glial Actin-GFP distribution and the overall effects on glial cell migration and morphological development were assessed. We found that RhoA and Rac1 have distinct roles in peripheral glial cell migration and nerve ensheathement; however, Cdc42 does not have a significant role in peripheral glial development. RhoA and Rac1 gain-of-function and loss-of-function mutants had both disruption of glial cell development and secondary effects on sensory axon fasciculation. Together, Actin cytoskeletal dynamics is an integral part of peripheral glial migration and nerve ensheathement, and is mediated by RhoA and Rac1. PMID- 12642489 TI - The role of the zebrafish nodal-related genes squint and cyclops in patterning of mesendoderm. AB - Nodal signals, a subclass of the TGFbeta superfamily of secreted factors, induce formation of mesoderm and endoderm in vertebrate embryos. We have examined the possible dorsoventral and animal-vegetal patterning roles for Nodal signals by using mutations in two zebrafish nodal-related genes, squint and cyclops, to manipulate genetically the levels and timing of Nodal activity. squint mutants lack dorsal mesendodermal gene expression at the late blastula stage, and fate mapping and gene expression studies in sqt(-/-); cyc(+/+) and sqt(-/-); cyc(+/-) mutants show that some dorsal marginal cells inappropriately form hindbrain and spinal cord instead of dorsal mesendodermal derivatives. The effects on ventrolateral mesendoderm are less severe, although the endoderm is reduced and muscle precursors are located nearer to the margin than in wild type. Our results support a role for Nodal signals in patterning the mesendoderm along the animal vegetal axis and indicate that dorsal and ventrolateral mesoderm require different levels of squint and cyclops function. Dorsal marginal cells were not transformed toward more lateral fates in either sqt(-/-); cyc(+/-) or sqt(-/-); cyc(+/+) embryos, arguing against a role for the graded action of Nodal signals in dorsoventral patterning of the mesendoderm. Differential regulation of the cyclops gene in these cells contributes to the different requirements for nodal related gene function in these cells. Dorsal expression of cyclops requires Nodal dependent autoregulation, whereas other factors induce cyclops expression in ventrolateral cells. In addition, the differential timing of dorsal mesendoderm induction in squint and cyclops mutants suggests that dorsal marginal cells can respond to Nodal signals at stages ranging from the mid-blastula through the mid gastrula. PMID- 12642490 TI - A homeobox gene, pnx, is involved in the formation of posterior neurons in zebrafish. AB - A homeobox gene, pnx, is expressed in prospective posterior neurogenic regions and later in primary neurons. pnx expression was regulated by a signal from the non-axial mesendoderm and by Notch signaling. Pnx contains an Eh1 repressor domain, which interacted with Groucho and acted as a transcriptional repressor. Misexpression of pnx increased neural precursor cells and postmitotic neurons, which express neurogenin1 and elavl3/HuC, respectively. Expression of an antimorphic Pnx (VP16Pnx) or inhibition of Pnx by antisense morpholino oligonucleotide led to the reduction in the number of a subset of primary neurons. Misexpression of pnx promoted neurogenesis independent of Notch signaling. Epistatic analyses showed that Pnx also functions downstream of the Notch signal. These data indicate that pnx is a novel repressor-type homeobox gene that regulates posterior neurogenesis. PMID- 12642491 TI - Joint regulation of the MAP1B promoter by HNF3beta/Foxa2 and Engrailed is the result of a highly conserved mechanism for direct interaction of homeoproteins and Fox transcription factors. AB - The MAP1B (Mtap1b) promoter presents two evolutionary conserved overlapping homeoproteins and Hepatocyte nuclear factor 3beta (HNF3beta/Foxa2) cognate binding sites (defining putative homeoprotein/Fox sites, HF1 and HF2). Accordingly, the promoter domain containing HF1 and HF2 is recognized by cerebellum nuclear extracts containing Engrailed and Foxa2 and has regulatory functions in primary cultures of embryonic mesmetencephalic nerve cells. Transfection experiments further demonstrate that Engrailed and Foxa2 interact physiologically in a dose-dependent manner: Foxa2 antagonizes the Engrailed driven regulation of the MAP1B promoter, and vice versa. This led us to investigate if Engrailed and Foxa2 interact directly. Direct interaction was confirmed by pull-down experiments, and the regions participating in this interaction were identified. In Foxa2 the interacting domain is the Forkhead box DNA-binding domain. In Engrailed, two independent interacting domains exist: the homeodomain and a region that includes the Pbx-binding domain. Finally, Foxa2 not only binds Engrailed but also Lim1, Gsc and Hoxa5 homeoproteins and in the four cases Foxa2 binds at least the homeodomain. Based on the involvement of conserved domains in both classes of proteins, it is proposed that the interaction between Forkhead box transcription factors and homeoproteins is a general phenomenon. PMID- 12642492 TI - Strabismus requires Flamingo and Prickle function to regulate tissue polarity in the Drosophila eye. AB - Tissue polarity in Drosophila is regulated by a number of genes that are thought to function in a complex, many of which interact genetically and/or physically, co-localize, and require other tissue polarity proteins for their localization. We report the enhancement of the strabismus tissue polarity phenotype by mutations in two other tissue polarity genes, flamingo and prickle. Flamingo is autonomously required for the establishment of ommatidial polarity. Its localization is dynamic throughout ommatidial development and is dependent on Frizzled and Notch. Flamingo and Strabismus co-localize for several rows posterior to the morphogenetic furrow and subsequently diverge. While neither of these proteins is required for the other's localization, Prickle localization is influenced by Strabismus function. Our data suggest that Strabismus, Flamingo and Prickle function together to regulate the establishment of tissue polarity in the Drosophila eye. PMID- 12642493 TI - Directional guidance of interneuron migration to the cerebral cortex relies on subcortical Slit1/2-independent repulsion and cortical attraction. AB - Tangential migration from the basal telencephalon to the cortex is a highly directional process, yet the mechanisms involved are poorly understood. Here we show that the basal telencephalon contains a repulsive activity for tangentially migrating cells, whereas the cerebral cortex contains an attractive activity. In vitro experiments demonstrate that the repulsive activity found in the basal telencephalon is maintained in mice deficient in both Slit1 and Slit2, suggesting that factors other than these are responsible for this activity. Correspondingly, in vivo analysis demonstrates that interneurons migrate to the cortex in the absence of Slit1 and Slit2, or even in mice simultaneously lacking Slit1, Slit2 and netrin 1. Nevertheless, loss of Slit2 and, even more so, Slit1 and Slit2 results in defects in the position of other specific neuronal populations within the basal telencephalon, such as the cholinergic neurons of the basal magnocellular complex. These results demonstrate that whereas Slit1 and Slit2 are not necessary for tangential migration of interneurons to the cortex, these proteins regulate neuronal migration within the basal telencephalon by controlling cell positioning close to the midline. PMID- 12642494 TI - Molecular regionalization of the neocortex is disrupted in Fgf8 hypomorphic mutants. AB - The neocortex is divided into multiple areas with specific architecture, molecular identity and pattern of connectivity with the dorsal thalamus. Gradients of transcription factor expression in the cortical primordium regulate molecular regionalization and potentially the patterning of thalamic projections. We show that reduction of Fgf8 levels in hypomorphic mouse mutants shifts early gradients of gene expression rostrally, thereby modifying the molecular identity of rostral cortical progenitors. This shift correlates with a reduction in the size of a molecularly defined rostral neocortical domain and a corresponding rostral expansion of more caudal regions. Despite these molecular changes, the topography of projections between the dorsal thalamus and rostral neocortex in mutant neonates appears the same as the topography of wild-type littermates. Overall, our study demonstrates the role of endogenous Fgf8 in regulating early gradients of transcription factors in cortical progenitor cells and in molecular regionalization of the cortical plate. PMID- 12642495 TI - Ssdp proteins bind to LIM-interacting co-factors and regulate the activity of LIM homeodomain protein complexes in vivo. AB - LIM-homeodomain transcription factors control a variety of developmental processes, and are assembled into functional complexes with the LIM-binding co factor Ldb1 (in mouse) or Chip (in Drosophila). We describe the identification and characterization of members of the Ssdp family of proteins, which we show to interact with Ldb1 and Chip. The N terminus of Ssdp is highly conserved among species and binds a highly conserved domain within Ldb1/Chip that is distinct from the domains required for LIM binding and self-dimerization. In Drosophila, Ssdp is expressed in the developing nervous system and imaginal tissues, and it is capable of modifying the in vivo activity of complexes comprised of Chip and the LIM-homeodomain protein Apterous. Null mutations of the ssdp gene are cell lethal in clones of cells within the developing wing disc. However, clones mutant for a hypomorphic allele give rise to ectopic margins, wing outgrowth and cell identity defects similar to those produced by mutant clones of Chip or apterous. Ssdp and Ldb/Chip each show structural similarity to two Arabidopsis proteins that cooperate with one another to regulate gene expression during flower development, suggesting that the molecular interactions between Ssdp and Ldb/Chip proteins are evolutionarily ancient and supply a fundamental function in the regulated control of transcription. PMID- 12642496 TI - Bazooka is a permissive factor for the invasive behavior of discs large tumor cells in Drosophila ovarian follicular epithelia. AB - Drosophila Bazooka and atypical protein kinase C are essential for epithelial polarity and adhesion. We show here that wild-type bazooka function is required during cell invasion of epithelial follicle cells mutant for the tumor suppressor discs large. Clonal studies indicate that follicle cell Bazooka acts as a permissive factor during cell invasion, possibly by stabilizing adhesion between the invading somatic cells and their substratum, the germline cells. Genetic epistasis experiments demonstrate that bazooka acts downstream of discs large in tumor cell invasion. In contrast, during the migration of border cells, Bazooka function is dispensable for cell invasion and motility, but rather is required cell-autonomously in mediating cell adhesion within the migrating border cell cluster. Taken together, these studies reveal Bazooka functions distinctly in different types of invasive behaviors of epithelial follicle cells, potentially by regulating adhesion between follicle cells or between follicle cells and their germline substratum. PMID- 12642497 TI - Two tcf3 genes cooperate to pattern the zebrafish brain. AB - Caudalizing factors operate in the context of Wnt/beta-catenin signaling to induce gene expression in discrete compartments along the rostral-caudal axis of the developing vertebrate nervous system. In zebrafish, basal repression of caudal genes is achieved through the function of Headless (Hdl), a Tcf3 homolog. In this study, we show that a second Tcf3 homolog, Tcf3b, limits caudalization caused by loss of Hdl function and although this Lef/Tcf family member can rescue hdl mutants, Lef1 cannot. Wnts can antagonize repression mediated by Tcf3 and this derepression is dependent on a Tcf3 beta-catenin binding domain. Systematic changes in gene expression caused by reduced Tcf3 function help predict the shape of a caudalizing activity gradient that defines compartments along the rostral caudal axis. In addition, Tcf3b has a second and unique role in the morphogenesis of rhombomere boundaries, indicating that it controls multiple aspects of brain development. PMID- 12642499 TI - Xbra functions as a switch between cell migration and convergent extension in the Xenopus gastrula. AB - During Xenopus gastrulation, the dorsal mesoderm exhibits two different cell behaviors in two different regions: active cell migration of prechordal mesoderm and convergent extension of chordamesoderm. Although many genes involved in specification and differentiation of the dorsal mesoderm have been studied, the role of these genes in controlling cell behaviors is poorly understood. To understand better the link between the development and cell behaviors of the dorsal mesoderm, we have examined these behaviors in dissociated cells and explants, where activin protein can induce both active cell migration and convergent extension. We find that Xbra, a transcription factor necessary for convergent extension, actively inhibits cell migration, both in animal cap explant assays and in the endogenous dorsal mesoderm. In addition, Xbra appears to inhibit cell migration by inhibiting adhesion to fibronectin. We propose that Xbra functions as a switch to keep cell migration and convergent extension as mutually exclusive behaviors during gastrulation. PMID- 12642498 TI - Zic1 represses Math1 expression via interactions with the Math1 enhancer and modulation of Math1 autoregulation. AB - Math1 is a basic helix-loop-helix transcription factor expressed in progenitor cells that give rise to dorsal commissural interneurons in the spinal cord, granule cells of the cerebellum, and sensory cells in the inner ear and skin. Transcriptional regulation of this gene is tightly controlled both temporally and spatially during nervous system development. The signals that mediate this regulation are likely integrated at the Math1 enhancer, which is highly conserved among vertebrate species. We have identified the zinc-finger transcription factor Zic1 as a regulator of Math1 expression. Zic1 binds a novel conserved site within the Math1 enhancer, and represses both the expression of endogenous Cath1 (chicken homolog of Math1) and the activity of a Math1 enhancer driven lacZ reporter when expressed in chick neural tubes. Repression by Zic1 blocks the autoregulatory activity of Math1 itself. Although previous reports have shown that Zic1 and Math1 are both induced by BMP signaling, these genes appear to have opposing functions, as Math1 acts to promote neuronal differentiation in the chick neural tube and excess Zic1 appears to block differentiation. Zic1-mediated repression of Cath1 transcription may modulate the temporal switch between the progenitor state and differentiating dorsal cell types during neural tube development. PMID- 12642500 TI - A Notch-independent function of Suppressor of Hairless during the development of the bristle sensory organ precursor cell of Drosophila. AB - Su(H)/CBF1 is a key component of the evolutionary conserved Notch signalling pathway. It is a transcription factor that acts as a repressor in the absence of the Notch signal. If Notch signalling is activated, it associates with the released intracellular domain of the Notch receptor and acts as an activator of transcription. During the development of the mechanosensory bristles of Drosophila, a selection process called lateral inhibition assures that only a few cells are selected out of a group to become sensory organ precursors (SOP). During this process, the SOP cell is thought to suppress the same fate in its surrounding neighbours via the activation of the Notch/Su(H) pathway in these cells. We show that, although Su(H) is required to prevent the SOP fate during lateral inhibition, it is also required to promote the further development of the SOP once it is selected. Importantly, in this situation Su(H) appears to act independently of the Notch signalling pathway. We find that loss of Su(H) function leads to an arrest of SOP development because of the loss of sens expression in the SOP. Our results suggest that Su(H) acts as a repressor that suppresses the activity of one or more negative regulator(s) of sens expression. We show that this repressor activity is encoded by one or several genes of the E(spl)-complex. Our results further suggest that the position of the SOP in a proneural cluster is determined by very precise positional cues, which render the SOP insensitive to Dl. PMID- 12642501 TI - LvTbx2/3: a T-box family transcription factor involved in formation of the oral/aboral axis of the sea urchin embryo. AB - T-box family transcription factors have been identified in many organisms and are frequently associated with patterning events during embryonic development. With an interest in the molecular basis of patterning in the sea urchin embryo, we identified several members of the T-box family in Lytechinus variegatus. Here, we report the cloning and characterization of an ortholog of the Tbx2/3 subfamily, LvTbx2/3. To characterize the spatial distribution of LvTbx2/3 protein throughout sea urchin embryogenesis, a polyclonal antiserum was generated. Nuclear localization of LvTbx2/3 initiated at the mesenchyme blastula stage and protein was present into the pluteus stage. Localization was asymmetric throughout this period and costaining with marker genes indicated that asymmetry was about the oral/aboral (O/A) axis. Asymmetric distribution of LvTbx2/3 was observed in the aboral territories of all three germ layers. In the skeletogenic mesoderm lineage, LvTbx2/3 expression was dynamic because expression appeared initially in all skeletogenic mesenchyme cells (PMCs) but, subsequently, became refined solely to the aboral ones during skeletogenesis. To determine if the aboral expression of LvTbx2/3 is linked between germ layers, and to place LvTbx2/3 in the sequence of events that specifies the O/A axis, the effects of a series of perturbations to O/A polarity on LvTbx2/3 expression in each germ layer were examined. Preventing the nuclear localization of beta-catenin, pharmacological disruption of the O/A axis with NiCl(2), overexpression of BMP2/4 and disruption of the extracellular matrix all blocked LvTbx2/3 expression in all germ layers. This indicates that expression of LvTbx2/3 in the aboral territories of each germ layer is a common aspect of O/A specification, downstream of the molecular events that specify the axis. Furthermore, blocking the nuclear localization of beta catenin, overexpression of BMP2/4 and disruption of the extracellular matrix also prevented the oral (stomodael) expression of LvBrachyury (LvBrac) protein, indicating that the O/A axis is established by a complex series of events. Last, the function of LvTbx2/3 in the formation of the O/A axis was characterized by examining the phenotypic consequences of ectopic expression of LvTbx2/3 mRNA on embryonic development and the expression of marker genes that identify specific germ layers and tissues. Ectopic expression of LvTbx2/3 produced profound morphogenetic defects in derivatives of each germ layer with no apparent loss in specification events in those tissues. This indicates that LvTbx2/3 functions as a regulator of morphogenetic movements in the aboral compartments of the ectoderm, endoderm and mesoderm. PMID- 12642502 TI - Impaired meiotic DNA-damage repair and lack of crossing-over during spermatogenesis in BRCA1 full-length isoform deficient mice. AB - Breast tumor suppressor gene 1 (BRCA1) plays an essential role in maintaining genomic integrity. Here we show that mouse Brca1 is required for DNA-damage repair and crossing-over during spermatogenesis. Male Brca1(Delta11/Delta11)p53(+/-) mice that carried a homozygous deletion of Brca1 exon 11 and a p53 heterozygous mutation had significantly reduced testicular size and no spermatozoa in their seminiferous tubules. During spermatogenesis, homologous chromosomes from the mutant mice synapsed and advanced to the pachytene stage but failed to progress to the diplotene stage. Our analyses revealed that the Brca1 mutation affected cellular localization of several DNA damage-repair proteins. This included prolonged association of gammaH2AX with sites of DNA damage, reduced sex body formation, diminished Rad51 foci and absence of Mlh1 foci in the pachytene stage. Consequently, chromosomes from mutant mice did not form chiasmata, a point that connects exchanging homologous chromosomes. Brca1-mutant spermatocytes also exhibited decreased RNA expression levels of several genes that are involved in DNA-damage repair, including RuvB like DNA helicase, XPB, p62 and TFIID. Of note, the premature termination of spermatogenesis at the pachytene stage was accompanied by increased apoptosis by both p53-dependent and p53-independent mechanisms. Thus, our study revealed an essential role of Brca1 in DNA-damage repair and crossing-over of homologous chromosomes during spermatogenesis. PMID- 12642503 TI - Lack of pendrin expression leads to deafness and expansion of the endolymphatic compartment in inner ears of Foxi1 null mutant mice. AB - Mice that lack the winged helix/forkhead gene Foxi1 (also known as Fkh10) are deaf and display shaker/waltzer behavior, an indication of disturbed balance. While Foxi1 is expressed in the entire otic vesicle at E9.5, it becomes gradually restricted to the endolymphatic duct/sac epithelium and at E16.5 Foxi1 expression in the inner ear is confined to this epithelium. Histological sections, paintfill experiments and whole-mount hybridizations reveal no abnormality in inner ear development of Foxi1(-/-) mice before E13.5. Between E13.5 and E16.5 the membranous labyrinth of inner ears from null mutants starts to expand as can be seen in histological sections, paint-fill experiments and three-dimensional reconstruction. Postnatally, inner ears of Foxi1(-/-) mice are extremely expanded, and large irregular cavities, compressing the cerebellum and the otherwise normal middle ear, have replaced the delicate compartments of the wild type inner ear. This phenotype resembles that of the human sensorineural deafness syndrome Pendred syndrome, caused by mutations in the PDS gene. In situ hybridization of Foxi1(-/-) endolymphatic duct/sac epithelium shows a complete lack of the transcript encoding the chloride/iodide transporter pendrin. Based on this, we would like to suggest that Foxi1 is an upstream regulator of pendrin and that the phenotype seen in Foxi1 null mice is, at least in part, due to defective pendrin-mediated chloride ion resorption in the endolymphatic duct/sac epithelium. We show that this regulation could be mediated by absence of a specific endolymphatic cell type--FORE (forkhead related) cells--expressing Foxi1, Pds, Coch and Jag1. Thus, mutations in FOXI1 could prove to cause a Pendred syndrome-like human deafness. PMID- 12642504 TI - Alterations in G protein and MAP kinase signaling pathways during cardiac remodeling in hypertension and heart failure. AB - The present study was undertaken to elucidate the G-protein and mitogen-activated kinase (MAP kinase) coupled signaling profile in a genetic model of hypertension and congestive heart failure (CHF) that mimics similar disease in humans. At the receptor level, Ang II type 1 receptor (AT1R) increased in left ventricular hypertrophy (LVH) and reverted to normal in CHF, whereas there was a downregulation of the Ang II type 2 receptor (AT2R) in CHF. At the transducer level, Galphaq and Galpha12 protein levels were unchanged during LVH but decreased significantly in CHF. In contrast, Gbeta and Galpha13 protein content were markedly upregulated in CHF. Furthermore, using phospho-specific antibodies in Western blots and in vitro kinase assays, we found at the effector level an upregulation of the small G-protein Rac1 activity during LVH but a decrease during CHF. In parallel, small G-protein Rho activity was significantly increased during LVH but was unchanged in failure. We found at the downstream level that MAP kinase isoforms extracellular signal regulated-kinase (ERK1/2), big mitogen activated kinase (BMK1/ERK5), C-jun N-terminal-activated kinase (JNKs/SAPKs), and stress-activated kinase (p38) bioactivities were increased during LVH. During CHF, ERK1/2 and JNK1/2 kinase activities were decreased, whereas BMK1/ERK5 kinase activity reverted to normal values. In conclusion, this study demonstrates, for the first time, multistep alterations of G-protein and MAP kinase signaling pathways in LVH and progression to failure in a genetic model of hypertension and failure. PMID- 12642505 TI - Differential expression of AT1 receptors in the pituitary and adrenal gland of SHR and WKY. AB - The renin-angiotensin (ANG) system has been implicated in the development of hypertension in spontaneously hypertensive rats (SHR). Because SHR are more susceptible to stress than normotensive Wistar-Kyoto rats (WKY), we measured the mRNA expression of AT1A, AT1B, and AT2 receptors in the hypothalamo-pituitary adrenal (stress) axis of male SHR in comparison to age-matched WKY at prehypertensive (3 to 4 weeks), developing (7 to 8 weeks), and established (12 to 13 weeks) stages of hypertension. AT1A receptor mRNA was mainly expressed in the hypothalamus and adrenal gland. AT1B receptor mRNA was detected in the pituitary and adrenal gland. AT2 receptor mRNA was prominent only in the adrenal gland. When compared with WKY, SHR showed increased AT1A receptor mRNA levels in the pituitary gland at all ages in contrast to reduced pituitary AT1B receptor mRNA levels. In the adrenal gland of SHR, AT1B receptor mRNA levels were decreased at the hypertensive stages when compared with WKY. The reduced expression of adrenal AT1B receptor mRNA was localized selectively in the zona glomerulosa by in situ hybridization. No differences were observed between WKY and SHR in the expression of hypothalamic ANG receptors. ANG significantly increased plasma levels of adrenocorticotropic hormone (ACTH) and corticosterone in dexamethasone-treated SHR but not in WKY. The aldosterone response to ANG was similar in SHR and WKY. Our results suggest a differential gene expression of AT1A and AT1B receptors in the hypothalamo-pituitary-adrenal axis of SHR and normotensive WKY and imply the participation of AT1 receptors in an exaggerated endocrine stress response of SHR to ANG. PMID- 12642506 TI - Mechanical stretch-induced apoptosis in smooth muscle cells is mediated by beta1 integrin signaling pathways. AB - Recently we demonstrated that mechanical stress induces apoptosis of vascular smooth muscle cells in vitro and in vein grafts (Mayr et al. FASEB J. 2000;15:261 270). The current study was designed to investigate molecular mechanisms of mechanical stretch-induced apoptosis. Smooth muscle cells cultivated on silicone elastomer plates precoated with collagen I, elastin, laminin, or Pronectin were subjected to cyclic mechanical stretch. Interestingly, in response to mechanical stress, the number of apoptotic cells increased significantly in cells growing on collagen I-coated plates but not on other matrixes. We therefore thought that receptors mediating binding to collagen I, such as integrin beta1 containing receptors, might be involved in signaling pathways leading to stretch-induced apoptosis. On collagen plates, mechanical stress rapidly activated p38 MAPK that phosphorylated p53 in smooth muscle cells. Lack of functional Rac completely abrogated p38 MAPK-p53 activation as well as apoptosis. Furthermore, mechanical stress resulted in increases of both integrin beta1 protein expression and activity as identified by Western blotting and Shc immunoprecipitation assays. Treatment with a beta1-integrin-blocking antibody or integrin signaling inhibitor cytochalasin B but not growth factor receptor inhibitor suramin abrogated both stretch-induced phosphorylation of p38 MAPK and p53 expression. Akin to the inhibition of p38 MAPK-p53 signaling, pretreatment with a beta1-integrin-blocking antibody or cytochalasin B but not suramin inhibited stretch-induced apoptosis on collagen plates. These results suggest that mechanical stress-induced apoptosis in vascular smooth muscle cells is mediated by beta1-integrin-rac-p38-p53 signaling pathways. PMID- 12642507 TI - Parallel renal and extremity blood supply abnormalities in nonmodulation: responses to ACE inhibition. AB - The aim of this work was to ascertain, in nonmodulating essential hypertension, whether the abnormality in the renal blood supply is extended to the extremities and showed a similar response to ACE inhibition and whether these abnormalities could be identified in normotensive offspring of hypertensives, as non-modulation is a familial process with genetic underpinnings. We measured forearm vascular blood flow (FBF) and forearm vascular resistance (FVR) by plethysmography and urinary albumin excretion in 20 normotensive without family story of hypertension (NT: 25+/-9 years), 10 modulating offspring of hypertensive parents (MHO: 25+/-6 years), 10 nonmodulating offspring of hypertensive parents (NMHO: 26+/-5 years), 12 modulating essential hypertensives (MHT: 34+/-5 years), and 11 nonmodulating essential hypertensives (NMHT: 32+/-4 years). Measurements were repeated in hypertensives after 3-month treatment with ramipril (5 mg daily). Nonmodulating individuals showed lower maximum FBF (NMHT: 41.96+/-3.3 mL/100 g per minute and NMHO: 35.6+/-9.0 mL/100 g per minute) than modulating subjects (MHT: 57.5+/-10.0 mL/100 g per minute and MHO: 51.8+/-7.0 mL/100 g per minute; P<0.003). Likewise, all nonmodulating subjects showed higher minimum FVR (NMHT: 2.5+/-0.2 AU; NMO: 2.8+/-0.5 AU) than modulating individuals (MHT: 1.9+/-0.5 AU; MHO, 1.8+/-0.3AU; P<0.025). Urinary albumin excretion was higher in NMHT and NMHO than MHT, MHO, and NT (P<0.05). Ramipril increased maximum FBF to 53.8+/-8.0 mL/100 g per minute and reduced minimum FVR to 1.9+/-0.5 AU in NMHT (P<0.01). Likewise, ramipril increased effective renal plasma flow and reduced renal vascular resistance and urinary albumin excretion only in NMHT (P<0.05). These results have shown an early involvement of the peripheral circulation in association with increased urinary albumin excretion not only in essential hypertensives but also in NMHO. The effectiveness of ramipril in reducing minimum FVR and urinary albumin excretion in NMHT also suggests a common mechanism. PMID- 12642508 TI - The role of Wnk4 in polygenic hypertension: a candidate gene analysis on rat chromosome 10. AB - Linkage analyses in experimental crosses of stroke-prone spontaneously hypertensive (SHRSP) and normotensive Wistar-Kyoto (WKY) rats have strongly suggested the presence of quantitative trait loci (QTL) influencing blood pressure and ACE levels on rat chromosome 10, which have been confirmed in multiple independent studies. Analysis of the orthologous region on human chromosome 17 also revealed significant linkage to blood pressure in several populations. Wnk4, a gene previously identified to cause pseudohypoaldosteronism type II, a rare mendelian form of arterial hypertension, is located on human chromosome 17. The hypothesis has been advanced that molecular variants of this gene might contribute to common polygenic forms of hypertension, since Wnk4 is located in a region of conserved synteny that demonstrates an overlap between quantitative trait loci for primary hypertension in humans and rats. In this report, we describe the confirmation of the blood pressure QTL on rat chromosome 10 by congenic approaches, spanning the Wnk4 locus. Comparative analysis of the complete coding sequence of Wnk4 in SHRSP and WKY strains revealed no mutation and demonstrated high conservation between rat and human proteins. Furthermore, comparison of mRNA levels in the kidney showed no differences between SHRSP and WKY. Additionally, we excluded a secondary effect of blood pressure on the transcriptional regulation of Wnk4. Our results fail to support a material contribution of Wnk4 to blood pressure regulation in this model of polygenic hypertension. Thus, Wnk4 is likely not to represent the underlying disease gene for the QTL captured in chromosome 10 congenic animals. PMID- 12642510 TI - Coronary myogenic constriction antagonizes EDHF-mediated dilation: role of KCa channels. AB - In hypertension, pressure-induced myogenic constriction and impaired endothelium derived hyperpolarizing factor (EDHF)-mediated dilation may contribute to increased vasomotor tone. Myogenic constriction as well as EDHF-mediated dilation may share common signaling mechanisms, and both may control KCa channel activity to set arterial tone. To investigate a potential relation between the 2 mechanisms, we studied coronary arteries of Sprague-Dawley rats for individual myogenic constriction compared with EDHF-mediated dilation of the same artery. EDHF-mediated dilation was measured as the maximal dilation to acetylcholine (100 micromol/L) after preconstriction, resistant to NO inhibition (NG-methyl-l arginine acetate salt, L-NMMA, 100 micromol/L), and prostaglandin inhibition (indomethacin, 10 micromol/L) but abolished by charybdotoxin (100 nmol/L) plus apamin (500 nmol/L). Individual coronary myogenic constriction at an intraluminal pressure of 70 mm Hg (n=9) ranged from 6% to 44% (24+/-4%). EDHF-mediated dilation ranged from 18% to 84% (42+/-7%). Elevating pressure to 130 mm Hg (n=8) increased myogenic constriction by 2-fold (P<0.01) and decreased EDHF-mediated dilation by 2.6-fold (P<0.01). Interestingly, individual myogenic constriction inversely correlated to individual EDHF-mediated dilation (r=-0.75, P<0.001, n=17). Pretreatment with the KCa channel opener NS1619 (30 micromol/L) prevented coronary myogenic constriction and increased EDHF-mediated dilation by 2.2-fold (P<0.01), whereas the KATP channel opener cromakalim (3 micromol/L) had no effect on EDHF-mediated dilation. For comparison, in mesenteric arteries (at 70 mm Hg) low myogenic constriction (2+/-1%) was associated with high EDHF-mediated dilation (93+/-2%), and pretreatment with NS1619 had no effect. Our results demonstrate that myogenic constriction in coronary arteries antagonizes EDHF mediated dilation. Activation of KCa channels with NS1619 reduces myogenic constriction and profoundly increases EDHF-mediated dilation, specifically in coronary arteries, suggesting a potential therapeutic impact to reduce coronary risk in hypertension. PMID- 12642509 TI - Calcium antagonist treatment by lercanidipine prevents hyperpolarization in essential hypertension. AB - Essential hypertension is associated with impaired endothelium-dependent vasodilation caused by oxidative stress-induced nitric oxide (NO) breakdown and compensatory production of a hyperpolarizing factor. To test whether calcium antagonist treatment can restore NO availability and prevent hyperpolarization through antioxidant properties, in 15 healthy subjects and 15 patients with essential hypertension, we studied forearm blood flow (strain-gauge plethysmography) modifications induced by intrabrachial bradykinin (5, 15, 50 ng/100 mL per minute), an endothelium-dependent vasodilator, in basal conditions, during infusion of NG-monomethyl-l-arginine (L-NMMA, 100 microg/100 mL per minute), an NO-synthase inhibitor, and ouabain (0.72 microg/100 mL per minute), an Na+-K+ ATPase inhibitor to prevent hyperpolarization. These infusions were repeated in the presence of the antioxidant vitamin C (8 mg/100 mL/min). The response to sodium nitroprusside was also evaluated. In controls, vasodilation to bradykinin was inhibited by L-NMMA and remained unchanged by ouabain or vitamin C. In hypertensive patients, vasodilation to bradykinin was blunted and resistant to L-NMMA but sensitive to ouabain. Vitamin C increased the response to bradykinin and restored the inhibiting effect of L-NMMA while preventing the effect of ouabain. In hypertensive patients, infusions were repeated after 3 month treatment with lercanidipine (10 to 20 mg daily). Lercanidipine decreased plasma lipoperoxides, isoprostanes, and malondialdehyde and increased plasma antioxidant capacity. Moreover, lercanidipine increased the vasodilation to bradykinin and restored the inhibiting effect of L-NMMA on bradykinin-induced vasodilation while preventing the effect of ouabain. Finally, vitamin C no longer exerted its facilitating activity. These results indicate that in essential hypertension, lercanidipine increases endothelium-dependent vasodilation by restoring NO availability and preventing hyperpolarization, an effect probably determined by antioxidant activity. PMID- 12642511 TI - Ral GDP dissociation stimulator and Ral GTPase are involved in myocardial hypertrophy. AB - Ras-related GTPase (Ral) is converted to the GTP-bound form by Ral GDP dissociation stimulator (Ral-GDS), a putative effector protein of Ras. Although a number of studies indicate that Ras induces cardiac hypertrophy, the functional role of Ral-GDS/Ral signaling pathway is as yet unknown in cardiac myocytes. We investigated the role of the Ral-GDS/Ral pathway in cardiac hypertrophy. Transfection of Ral-GDS and constitutively active mutant of Ral (RalG23V) in cultured rat neonatal myocytes stimulated promoter activity of c-fos (5.4-fold and 2.6-fold, P<0.01), alpha-skeletal actin (2.7-fold and 2.1-fold, P<0.01), and beta-myosin heavy chain-luciferase (2.8-fold and 2.3-fold, P<0.01). Ral-GDS induced or RalG23V-induced promoter activation was increased synergistically with activated Ras (RasG12V). Dominant-negative mutant of Ral (RalS28N) partially inhibited RasG12V induced promoter activation. Cardiac myocytes transfected with RalG23V showed increased cell size compared with nontransfected or vector transfected cells (2.1-fold, P<0.01). Cardiotrophin-1 (CT-1) upregulated Ral-GDS mRNA expression and induced Ral activation. CT-1-induced Ral-GDS mRNA expression was inhibited by overexpression of the dominant-negative mutant of STAT3. Moreover, Ral activity was elevated in hypertrophied hearts (2.1-fold, P<0.01) by mechanical stress in association with increased CT-1 expression and signal transducer and activator of transcription 3 (STAT3) phosphorylation in the rat aortic banding model. Ral-GDS/Ral pathway is involved in a wide range of gene expressions and is activated by hypertrophic stimuli in vitro and in vivo. SATA3 may play a key role in Ral-GDS expression and Ral activation. Our data provide evidence that the Ral-GDS/Ral signaling pathway is a link to the process of cardiac hypertrophy. PMID- 12642512 TI - Dietary salt regulates renal SGK1 abundance: relevance to salt sensitivity in the Dahl rat. AB - Serum and glucocorticoid-induced kinase 1 (SGK1) activates the epithelial sodium channel (eNaC) in tubules. We examined renal SGK1 abundance in salt-adaptation and in salt-sensitive hypertension. Sprague-Dawley and Dahl salt-sensitive rats were placed on either 8% or 0.3% NaCl diets for 10 days. Plasma aldosterone levels were approximately 2.5-fold greater on 0.3% versus 8% NaCl diets in both rat strains. Both serum and glucocorticoid-induced kinase 1 transcript and protein abundance were less (P<0.01) in Sprague-Dawley rats and greater (P<0.01) in Dahl salt-sensitive rats on 8% versus 0.3% NaCl diets. The cDNA sequences of serum and glucocorticoid-induced kinase 1 in both strains of rat were the same. The present results provide evidence that the abundance of serum and glucocorticoid-induced kinase 1 in rat kidney may play a role in salt adaptation and the pathogenesis of hypertension and suggests that aldosterone is not the primary inducer of SGK1 in the Sprague-Dawley rat. PMID- 12642513 TI - Skillmix: an advance or an excuse? PMID- 12642514 TI - Reforming emergency care; for patients. PMID- 12642515 TI - Reforming the UK emergency care system. PMID- 12642516 TI - The advances and evidence base for prehospital care. PMID- 12642517 TI - Turbulent times. PMID- 12642518 TI - The NICE head injury guidelines. PMID- 12642519 TI - See and Treat: a management driven method of achieving targets or a tool for better patient care? One size does not fit all. PMID- 12642520 TI - Don't throw triage out with the bathwater. PMID- 12642521 TI - One size does not fit all. View 2. PMID- 12642522 TI - Clinical decision units in the emergency department: old concepts, new paradigms, and refined gate keeping. PMID- 12642523 TI - Designing the accident and emergency system: lessons from manufacturing. AB - OBJECTIVES: To review the literature on manufacturing process design and demonstrate applicability in health care. METHODS: Literature review and application of theory using two years activity data from two healthcare communities and extensive observation of activities over a six week period by seven researchers. RESULTS: It was possible to identify patient flows that could be used to design treatment processes around the needs of the patient. Some queues are built into existing treatment processes and can be removed by better process design. Capacity imbalance, not capacity shortage, causes some unnecessary waiting in accident and emergency departments. CONCLUSIONS: Clinicians would find that modern manufacturing theories produce more acceptable designs of systems. In particular, good quality is seen as a necessary pre requisite of fast, efficient services. PMID- 12642524 TI - Discharge from triage: modelling the potential in different types of emergency department. AB - OBJECTIVE: To assess the potential for patients to be assessed and discharged directly from triage in an emergency department (ED). METHODS: Modelling was undertaken by collection of retrospective electronic data from four different EDs. Serial removal of groups was undertaken using data from coding systems related to patients details of admission/treatment/investigations and procedure undertaken. The final group left were analysed for ambulance usage, prior primary care consultation, and age group. RESULTS: 29.4% patients were discharged after clinical assessment but without any specific treatment or investigation. It was seen that of the patients who can be considered for discharge from triage, 15.5% were brought to the ED by ambulance, 3.5% were patients who had already consulted primary care, and 11% were children. CONCLUSIONS: This study suggests that a large percentage of patients seen in EDs may not require the extra facilities of that department. There is potential for a large number to be discharged within a few minutes of arrival if appropriate assessment skills are available at first contact. This may require more senior assessment than is currently used. This study has not assessed safety of such a system or the times of day when it is best deployed. PMID- 12642525 TI - Social care's impact on emergency medicine: a model to test. AB - Mainly in response to the policy drive to avoid unnecessary acute hospital admissions and delayed discharge on social grounds, there has been a gradual development of social work services attached to emergency departments (EDs) in the UK. In the absence of a clearly articulated evidence base or debate about the roles of ED attached social workers, a model of ED based social work practice and indicative supporting evidence is presented. It is argued that social workers may be able to contribute to the efficiency and effectiveness of hospital services while providing a key point of access to social care services. A number of obstacles remain to the implementation of this model of service, including the narrow focus of current social care practice, the hours that a social work service is normally provided, chronic under-funding, and continuing perverse incentives in the health and social care system. More systematic evidence in the UK context is needed to support the case for change. PMID- 12642526 TI - Use of emergency observation and assessment wards: a systematic literature review. AB - INTRODUCTION: Observation and assessment wards allow patients to be observed on a short-term basis and permit patient monitoring and/or treatment for an initial 24 48 hour period. They should permit concentration of emergency activity and resources in one area, and so improve efficiency and minimise disruption to other hospital services. These types of ward go under a variety of names, including observation, assessment, and admission wards. This review aims to evaluate the current literature and discuss assessment/admission ward functionality in terms of organisation, admission criteria, special patient care, and cost effectiveness. METHODS: Search of the literature using the Medline and BIDS databases, combined with searches of web based resources. Critical assessment of the literature and the data therein is presented. RESULTS: The advantages and disadvantages of the use of assessment/admission wards were assessed from the current literature. Most articles suggest that these wards improve patient satisfaction, are safe, decrease the length of stay, provide earlier senior involvement, reduce unnecessary admissions, and may be particularly useful in certain diagnostic groups. A number of studies summarise their organisational structure and have shown that strong management, staffing, organisation, size, and location are important factors for efficient running. There is wide variation in the recommended size of these wards. Observation wards may produce cost savings largely relating to the length of stay in such a unit. CONCLUSION: All types of assessment/admission wards seem to have advantages over traditional admission to a general hospital ward. A successful ward needs proactive management and organisation, senior staff involvement, and access to diagnostics and is dependent on a clear set of policies in terms of admission and care. Many diagnostic groups benefit from this type of unit, excluding those who will inevitably need longer admission. Vigorous financial studies have yet to be undertaken in the UK. Definitions of observation, assessment, and admission ward are suggested. PMID- 12642528 TI - Can good bed management solve the overcrowding in accident and emergency departments? PMID- 12642527 TI - Analysis of patient flow in the emergency department and the effect of an extensive reorganisation. AB - OBJECTIVES: To evaluate the different internal factors influencing patient flow, effectiveness, and overcrowding in the emergency department (ED), as well as the effects of ED reorganisation on these indicators. METHODS: The study compared measurements at regular intervals of three hours of patient arrivals and patient flow between two comparable periods (from 10 February to 2 March) of 1999 and 2000. In between, a structural and staff reorganisation of ED was undertaken. The main reason for each patient remaining in ED was recorded and allocated to one of four groups: (1) factors related to ED itself; (2) factors related to ED-hospital interrelation; (3) factors related to hospital itself; and (4) factors related to neither ED nor hospital. The study measured the number of patients waiting to be seen and the waiting time to be seen as effectiveness markers, as well as the percentage of time that ED was overcrowded, as judged by numerical and functional criteria. RESULTS: Effectiveness of ED was closely related with some ED related and hospital related factors. After the reorganisation, patients who remained in ED because of hospital related or non-ED-non-hospital related factors decreased. ED reorganisation reduced the number of patients waiting to be seen from 5.8 to 2.5 (p<0.001) and waiting time from 87 to 24 minutes (p<0.001). Before the reorganisation, 31% and 48% of the time was considered to be overcrowded in numerical and functional terms respectively. After the reorganisation, these figures were reduced to 8% and 15% respectively (p<0.001 for both). CONCLUSIONS: ED effectiveness and overcrowding are not only determined by external pressure, but also by internal factors. Measurement of patient flow across ED has proved useful in detecting these factors and in being used to plan an ED reorganisation. PMID- 12642529 TI - Meeting National Service Framework goals for patients presenting with acute myocardial infarction. AB - BACKGROUND: The National Service Framework for coronary heart disease established clear standards for the management of patients with acute myocardial infarction in March 2000. This study evaluates an emergency department's thrombolysis performance in light of these standards. SETTING: Inner city teaching hospital emergency department. METHODS: The data were prospectively collected using a formal clinical pathway for all patients receiving thrombolysis in the emergency department between February 2000 and January 2001. Cases were reviewed at monthly multidisciplinary audit meetings. Regular feedback complemented routine teaching for both nursing and medical staff. RESULTS: 127 patients were thrombolysed, of whom 92 (72%) were immediately eligible. Some 77% of these had a door to needle time of less than 30 minutes and 38% less than 20 minutes. Twenty per cent of patients had a call to door time of less than 30 minutes. Some 84% of patients managed by the emergency department team had a door to needle time of less than 30 minutes compared with 53% of those patients seen by duty physicians. CONCLUSIONS: The thrombolysis target set by the National Service Framework for April 2002 is achievable. The target set for April 2003 remains an ambitious goal. Overall call to needle times are undermined by call to door times. Emergency department teams may be more efficient than duty physicians in processing patients needing thrombolysis. PMID- 12642530 TI - Emergency nurse practitioners: a three part study in clinical and cost effectiveness. AB - AIMS: To compare the clinical effectiveness and costs of minor injury services provided by nurse practitioners with minor injury care provided by an accident and emergency (A&E) department. METHODS: A three part prospective study in a city where an A&E department was closing and being replaced by a nurse led minor injury unit (MIU). The first part of the study took a sample of patients attending the A&E department. The second part of the study was a sample of patients from a nurse led MIU that had replaced the A&E department. In each of these samples the clinical effectiveness was judged by comparing the "gold standard" of a research assessment with the clinical assessment. Primary outcome measures were the number of errors in clinical assessment, treatment, and disposal. The third part of the study used routine data whose collection had been prospectively configured to assess the costs and cost consequences of both models of care. RESULTS: The minor injury unit produced a safe service where the total package of care was equal to or in some cases better than the A&E care. Significant process errors were made in 191 of 1447 (13.2%) patients treated by medical staff in the A&E department and 126 of 1313 (9.6%) of patients treated by nurse practitioners in the MIU. Very significant errors were rare (one error). Waiting times were much better at the MIU (mean MIU 19 minutes, A&E department 56.4 minutes). The revenue costs were greater in the MIU (MIU pound 41.1, A&E department pound 40.01) and there was a great difference in the rates of follow up and with the nurses referring 47% of patients for follow up and the A&E department referring only 27%. Thus the costs and cost consequences were greater for MIU care compared with A&E care (MIU pound 12.7 per minor injury case, A&E department pound 9.66 per minor injury case). CONCLUSION: A nurse practitioner minor injury service can provide a safe and effective service for the treatment of minor injury. However, the costs of such a service are greater and there seems to be an increased use of outpatient services. PMID- 12642531 TI - Designing a research project: randomised controlled trials and their principles. PMID- 12642532 TI - Seldinger technique chest drains and complication rate. AB - A short cut review was carried out to establish whether the seldinger "over the wire" technique is better than other techniques of pneumothorax drainage. Altogether 28 papers were found using the reported search, of which three presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 12642534 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Antacids and diagnosis in patients with atypical chest pain. AB - A short cut review was carried out to establish whether antacids can be used as a diagnostic test in atypical chest pain. Altogether 374 papers were found using the reported search, of which two presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 12642535 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Cervical spine imaging in children under 9 after trauma. AB - A short cut review was carried out to establish whether the odontoid peg view is useful to radiologically exclude cervical spine injury in children under 9 years of age. Altogether 156 papers were found using the reported search, of which two presented the best evidence to answer the clinical question. In addition recent guidelines are noted. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 12642536 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Need for cervical spine imaging for alert children after trauma. AB - A short cut review was carried out to establish whether clinical examination can be used to exclude cervical spine injury in alert children. Altogether 298 papers were found using the reported search, of which three presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 12642537 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Conservative management [correction of mangement] of asymptomatic cocaine body packers. AB - A short cut review was carried out to establish whether asymptomatic cocaine body packers can be managed conservatively. Altogether 171 papers were found using the reported search, of which four presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 12642538 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Acute myocardial infarction in cocaine induced chest pain presenting as an emergency. AB - A short cut review was carried out to establish the incidence of acute myocardial infarction in patients presenting as emergencies with post-cocaine chest pain. Altogether 198 papers were found using the reported search, of which eight presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 12642540 TI - Computer assisted assessment and advice for "non-serious" 999 ambulance service callers: the potential impact on ambulance despatch. AB - OBJECTIVE: To investigate the potential impact for ambulance services of telephone assessment and triage for callers who present with non-serious problems (Category C calls) as classified by ambulance service call takers. DESIGN: Pragmatic controlled trial. Calls identified using priority dispatch protocols as non-serious were allocated to intervention and control groups according to time of call. Ambulance dispatch occurred according to existing procedures. During intervention sessions, nurses or paramedics within the control room used a computerised decision support system to provide telephone assessment, triage and, if appropriate, offer advice to permit estimation of the potential impact on ambulance dispatch. SETTING: Ambulance services in London and the West Midlands. SUBJECTS: Patients for whom emergency calls were made to the ambulance services between April 1998 and May 1999 during four hour sessions sampled across all days of the week between 0700 and 2300. MAIN OUTCOME MEASURES: Triage decision, ambulance cancellation, attendance at an emergency department. RESULTS: In total, there were 635 intervention calls and 611 controls. Of those in the intervention group, 330 (52.0%) were triaged as not requiring an emergency ambulance, and 119 (36.6%) of these did not attend an emergency department. This compares with 55 (18.1%) of those triaged by a nurse or paramedic as requiring an ambulance (odds ratio 2.62; 95% CI 1.78 to 3.85). Patients triaged as not requiring an emergency ambulance were less likely to be admitted to an inpatient bed (odds ratio 0.55; 95% CI 0.33 to 0.93), but even so 30 (9.2%) were admitted. Nurses were more likely than paramedics to triage calls into the groups classified as not requiring an ambulance. After controlling for age, case mix, time of day, day of week, season, and ambulance service, the results of a logistic regression analysis revealed that this difference was significant with an odds ratio for nurses:paramedics of 1.28 (95% CI 1.12 to 1.47). CONCLUSIONS: The findings indicate that telephone assessment of Category C calls identifies patients who are less likely to require emergency department care and that this could have a significant impact on emergency ambulance dispatch rates. Nurses were more likely than paramedics to assess calls as requiring an alternative response to emergency ambulance despatch, but the extent to which this relates to aspects of training and professional perspective is unclear. However, consideration should be given to the acceptability, reliability, and cost consequences of this intervention before it can be recommended for full evaluation. PMID- 12642541 TI - Use of out of hours services: a comparison between two organisations. AB - OBJECTIVES: To investigate differences in numbers and characteristics of patients using primary or emergency care because of differences in organisation of out of hours care. BACKGROUND: Increasing numbers of self referrals at the accident and emergency (A&E) department cause overcrowding, while a substantial number of these patients exhibit minor injuries that can be treated by a general practitioner (GP). METHODS: Two different organisations of out of hours care in two Dutch cities (Heerlen and Maastricht) were investigated. Important differences between the two organisations are the accessibility and the location of primary care facility (GP cooperative). The Heerlen GP cooperative is situated in the centre of the city and is respectively 5 km and 9 km away from the two A&E departments situated in the area of Heerlen. This GP cooperative can only be visited by appointment. The Maastricht GP cooperative has free access and is located within the local A&E department. During a three week period all registration forms of patient contacts with out of hours care (GP cooperative and A&E department) were collected and with respect to the primary care patients a random sample of one third was analysed. RESULTS: For the Heerlen and Maastricht GP cooperative the annual contact rate, as extrapolated from our data, per 1000 inhabitants per year is 238 and 279 respectively (chi(2)((1df))=4.385, p=0.036). The contact rate at the A&E departments of Heerlen (n=66) and Maastricht (n=52) is not different (chi(2)((1df))=1.765, p=0.184). Some 51.7% of the patients attending the A&E department in Heerlen during out of hours were self referred, compared with 15.9% in Maastricht (chi(2)((1df))=203.13, p<0.001). CONCLUSIONS: The organisation of out of hours care in Maastricht has optimised the GP's gatekeeper function and thereby led to fewer self referrals at the A&E department, compared with Heerlen. PMID- 12642543 TI - The Surrey Emergency Care System: a countywide initiative for change. AB - Modernisation is at the core of government policy. Within the NHS it has variously focused on accident and emergency, coronary heart disease, clinical governance and information technology although each strand has tended to operate in isolation. The Surrey Emergency Care System is a programme combining the strands into a single countywide initiative and that lays the technical and clinical foundations of a future integrated unscheduled care network. This paper describes the programme, its potential impact, and offers some insight into the barriers to change that the project has met so far. PMID- 12642542 TI - Evaluation of emergency medical services systems: a classification to assist in determination of indicators. AB - Emergency medical services (EMS) systems, and prehospital care are difficult to evaluate. Accordingly, the true efficacy and value of such systems are difficult to determine. The multitude of variations and combinations of involved factors makes standardisation and comparison difficult, and universal indicators are hard to develop. Various attempts have been made to determine valid indicators of effectiveness, but there has been little success. Prehospital care has been seen by some as a single entity. As a result, experience from well resourced first world trauma centres has been taken, by many, to be applicable to all prehospital situations. This article attempts to assist in the development of valid EMS indicators of performance and effectiveness by categorising prehospital scenarios into a classification reflecting the reality of their conditions of practice. PMID- 12642545 TI - Would a prehospital practitioner model improve patient care in rural Australia? AB - BACKGROUND: Existing rural prehospital models have been criticised for being isolated from the healthcare system, and for following inflexible clinical protocols. Greater reliance on clinical judgement and informed decision making in the prehospital setting offer the potential to improve patient care. METHODS: Soft systems methodology was used to develop and critically appraise the prehospital practitioner model as an alternative to existing models. This approach started from the philosophical viewpoint that prehospital services should be patient centred. Soft systems methodology was used to structure the elements of prehospital systems and the relations between them into metaphors and pictures that could be analysed. RESULTS: This analysis showed that the most powerful reason for advocating the prehospital practitioner model is that it places prehospital systems within a symbiotic relationship with the healthcare system. Unlike the existing emergency service models or the "chain of survival" model, it is an integrated system that provides a range of services at multiple points during the patient care cycle. Thus, the prehospital practitioner would have roles in the prevention of injury and illness, responding to emergencies, facilitating recovery, and planning future strategies for a healthy community. CONCLUSIONS: Implementing this new model would see the prehospital system using its available capacity more effectively to fulfill broader public health and primary care outreach roles than is currently the case. Patients would be referred or transported to the most appropriate and cost effective facility as part of a seamless system that provides patients with well organised and high quality care. PMID- 12642544 TI - Developing a community paramedic practitioner intermediate care support scheme for older people with minor conditions. AB - INTRODUCTION: The Department of Health document Reforming emergency care stated that new initiatives need to be developed to improve the care and assessment of patients. The Audit Commission has suggested that ambulance services should be allowed to decide who should be sent to each type of emergency and treat some patients at home. AIMS: This scheme explores a new way of providing clinical assessment of older patients in their homes or in care homes within Sheffield. It sets out to provide a very patient centred model of care by providing community based clinical assessment for patients presenting to the emergency services with minor acute conditions. Scope, development, and structure of scheme: The scheme trains paramedic practitioners in the assessment and treatment of minor conditions to emergency nurse practitioner level. It consists of a three week full time theory based course and a 45 day period of supervised clinical practice based in the emergency department, minor injury unit, care of the elderly falls clinic, and with community services. Subsequently, the competence of the practitioners is assessed. Service delivery: The service will be activated by a 999 call between 0800 to 2000 each day. It is anticipated that between 25% to 50% of patients eligible to receive the service will be assessed and treated within the home. This approach to providing emergency care is untested and the frequency of use, patient acceptability, safety, and cost effectiveness are unknown, therefore rigorous assessment is essential through a randomised controlled trial. PMID- 12642546 TI - Effect of weather on attendance with injury at a paediatric emergency department. AB - OBJECTIVES: To ascertain whether the weather affects the attendance rate of children with injuries at a paediatric accident and emergency department. METHODS: The maximum daily temperature and weather conditions (rain/cloud/sun) were noted over a three month period in spring/summer 2002, together with the number of children attending with new injuries or trauma. RESULTS: There was a direct association between trauma attendance and clement weather with higher attendances on dry and sunny days. There was a less obvious association between maximum daily temperature and attendance. CONCLUSIONS: These findings confirm the anecdotal belief that warm sunny weather results in a higher attendance of paediatric injuries. PMID- 12642547 TI - Lesson learnt: don't prescribe heparin for hemoperitoneum! AB - This paper describes a case of spontaneous rupture of the spleen that was misdiagnosed as pulmonary embolism. Because of a lack of history of trauma, rarity of occurrence, and confusing clinical presentations, an unwary clinician may fail to diagnose this condition in time. It may result in erroneous treatment of a condition, which is potentially curable. PMID- 12642548 TI - National Service Framework fails to address the decision time. PMID- 12642549 TI - Expanding the precautionary principle. PMID- 12642550 TI - Injury prevention: blurring the distinctions between home and work. PMID- 12642551 TI - Diversity of trends in occupational injury mortality in the United States, 1980 96. AB - OBJECTIVES: Although the United States has generally enjoyed declining rates of fatal occupational injury, the rate of decline has not been uniform. To examine the heterogeneity of trends, changes in fatal occupational injury rates from 1980 to 1996 were estimated by occupation, industry, geographic region, and demographic group. METHODS: Deaths due to injury at work during 1980-96 were identified from the US National Traumatic Occupational Fatality database and populations at risk were estimated from the census of population. Mortality rates were computed for unintentional injuries, homicides, and all injuries combined. The annual rate of change was estimated using Poisson regression to model the death rate as a function of time. RESULTS: The estimated average rates for all fatal occupational injuries and for unintentional injuries declined by 3% per year, while the estimated rate of homicide declined <1% per year. The improvement was faster for men (3% per year) than for women (<1% per year) and for younger relative to older workers (7% per year v 2%-3% per year). Trends were also geographically heterogeneous, with the most rapid declines (7%-8% per year) in the South and West. Injury rates for most occupations and industries declined at near the average rate, but some experienced no change or an increase. The rate of homicide also increased in a number of occupations and industries. CONCLUSIONS: Broad downward trends in occupational fatality rates may be explained by several factors, including organized safety efforts, product and process changes, and the ongoing shift of employment toward safer sectors. Disparities in fatal injury trends draw attention to potential opportunities to reduce risk: work settings with increasing injury rates are of particular concern. PMID- 12642552 TI - Unintentional fatal injuries arising from unpaid work at home. AB - OBJECTIVE: Unpaid work in and around the home is a common and potentially high risk activity, yet there is limited information about the circumstances surrounding resulting injuries. This study aimed to describe circumstances surrounding fatal injuries resulting from home duties activities, in order to identify and prioritise areas for prevention. DESIGN AND SETTING: Coroners' reports on all unintentional deaths in Australia from 1989-92 inclusive were inspected to identify deaths of interest. Rates were calculated using population data and incorporating measures of time engaged in particular home duties activities. RESULTS: There were 296 home duties deaths over the four year period. Most (83%) deaths were of males, and males had 10 times the risk of fatal injury compared with females. The most common activities resulting in fatal injuries were home repairs, gardening, and car care. The highest risk activities (deaths per million persons per year per hour of activity) were home repairs (49), car care (20), home improvements (18), and gardening (16). Being hit by inadequately braced vehicles during car maintenance, falls from inadequately braced ladders, contact with fire and flames while cooking, and contact with electricity during maintenance were the most common injury scenarios. CONCLUSIONS: Fatal injury of persons engaged in unpaid domestic work activities is a significant cause of death. Use of activity specific denominator data allows appropriate assessment of the degree of risk associated with each activity. The recurrence of similar circumstances surrounding many independent fatal incidents indicates areas where preventative interventions might be usefully targeted. PMID- 12642554 TI - Seatbelt use by high school students. AB - OBJECTIVE: To determine seatbelt use of teenage drivers arriving at high schools in the morning and at evening football games compared with belt use of adults driving teenage passengers to these events, and teenage passenger belt use depending on whether they were being driven by another teenager or an adult. METHODS: Unobtrusive observations of belt use were made at 12 high schools in Connecticut and Massachusetts. RESULTS: Among males, teenage drivers had lower belt use than adults; differences between female teenage and female adult drivers were slight. Teenage passengers had lower belt use in vehicles driven by other teenagers than in cars driven by adults, but more than 40% of teenage passengers in vehicles driven by adults, presumed in most cases to be the teenager's parent, were not belted. Teenage passenger belt use was lower than teenage driver use regardless of gender. These differences were found both at morning arrivals and at football games, but teenage belt use was not much different in these two settings. Teenage passengers were belted more often if drivers were belted, whether the driver was another teenager or an adult, but a third of male passengers and 25%-30% of female passengers were unbelted even when drivers were belted. CONCLUSION: Teenagers have high crash risk but low belt use, which adds to their injury problem. Avenues to address this include strong belt use laws and their enforcement, building belt use requirements into graduated licensing systems, keeping young beginners out of high risk driving situations, and finding ways to influence parents and other adults to ensure that their teenage passengers use seatbelts. PMID- 12642553 TI - Nail gun injuries in residential carpentry: lessons from active injury surveillance. AB - OBJECTIVE: To describe circumstances surrounding injuries involving nail guns among carpenters, calculate injury rates, identify high risk groups and preventive measures. METHODS: and setting: Active injury surveillance was used to identify causes of injury among a large cohort of union residential and drywall carpenters. Injured carpenters were interviewed by experienced journeymen; enumeration of workers and hourworked were provided by the union. The combined data allowed definition of a cohort of carpenters, their hours worked, detailed information on the circumstances surrounding injuries, and identification of preventive measures from the perspectives of the injured worker and an experienced investigator. RESULTS: Nail guns were involved in 14% of injuries investigated. Ninety percent of these injuries were the result of the carpenter being struck, most commonly by a nail puncturing a hand or fingers. The injury rate among apprentices was 3.7 per 200 000 hours worked (95% confidence interval (CI) 2.7 to 4.9) compared with a rate of 1.2 among journeymen (95% CI 0.80 to 1.7). While not always the sole contributing factor, a sequential trigger would have likely prevented 65% of the injuries from tools with contact trip triggers. CONCLUSIONS: Training, engineering, and policy changes in the workplace and manufacturing arena are all appropriate targets for prevention of these injuries. Use of sequential triggers would likely decrease acute injury rates markedly. Over 70% of injuries among residential carpenters were associated with through nailing tasks (such as nailing studs or blocks, trusses or joists) or toe nailing (angled, corner nailing) as opposed to flat nailing used for sheathing activities; this provides some indication that contact trip tools could be used solely for flat nailing. PMID- 12642555 TI - Seatbelt legislation in Japan: high risk driver mortality and seatbelt use. AB - OBJECTIVES: To clarify why seatbelt legislation did not achieve the expected reduction in mortality in Japan. LOCATION AND BACKGROUND: Seatbelt legislation was enacted in Japan in September 1985 and penalties were introduced in November 1986. METHODS: The driver deaths per vehicle km traveled (D/VKT) were calculated to adjust for changes in traffic volume. Decreases in D/VKT were compared with the reduction expected after legislation. The association between percentage changes of driver D/VKT, seatbelt use rate, and seatbelt non-use rate were explored. Deaths of passengers, pedestrians, and cyclists were also examined. Mortality data were obtained from vital statistics, traffic volume figures from the Ministry of Land, Infrastructure, and Transport, and seatbelt use rates from the National Police Agency. RESULTS: Although the decrease in D/VKT after the law was enforced was larger than the absolute number of deaths, it was far less than predicted. The percentage decrease in seatbelt non-use rate showed the strongest correlation with the percentage decrease in driver mortality. Mortality did not increase among other road users after the law was enacted. CONCLUSION: Accurate evaluation of the effect of seatbelt legislation must take into account changes in traffic volume. The selective recruitment hypothesis-that high risk drivers were less responsive to seatbelt legislation-fits well with the findings. There was no conclusive evidence supporting risk compensation-that is, an increase in injuries among other road users. PMID- 12642556 TI - Socioeconomic variation in injury in children and older people: a population based study. AB - OBJECTIVES: To compare hospital admission rates for all causes and specific causes of injury in children and the elderly by a measure of economic deprivation. STUDY DESIGN: All emergency admissions for Welsh residents from 1997 99 were located to one of 865 electoral tracts, which were grouped into fifths using a measure of socioeconomic deprivation. Standardised admission rates for all ages and 0-14, 15-75, and 75+ year groups for each quintile were calculated with 95% confidence intervals. RESULTS: There were 90 935 admissions in a population of 2.84 million yielding a crude admission rate of 1601/100 000/year and a standardised rate of 1493/100 000. The ratio of admissions in deprived and affluent areas varied with category of injury and age group. In general, socioeconomic variations in injury rates were much smaller in older people than in children with the exception of pedestrian related injuries where the rates were similar. The largest variations were for injuries sustained in assaults or self inflicted. CONCLUSIONS: The relationship between socioeconomic position and injury varies by cause and age group. This should be considered when developing area based preventive interventions or monitoring the effectiveness of policies to reduce inequalities in injury occurrence. PMID- 12642557 TI - Discrepancy between actual and estimated speeds of drivers in the presence of child pedestrians. AB - OBJECTIVES: First, to measure the speeds of vehicles with and without children on the footpath, and second to compare these with drivers' estimates of how fast they would go in these conditions. DESIGN: The speeds of vehicles in three conditions: control (no children present), children playing with a ball on the footpath, and children waiting to cross the road, were measured using speed tubes during two 55 minute sessions. Drivers' estimates of their speeds were measured with a questionnaire. SETTING: Speeds were measured on a main road in Auckland, New Zealand. The questionnaire was conducted at another time with drivers stopping for petrol approximately 500 metres from the measurement site. SUBJECTS: A total of 1446 speed measurements were taken and 93 drivers' questionnaire responses were analysed. RESULTS: The mean free speed of vehicles in the control condition was 55.60 kph, with drivers' estimates being 56.37 kph. When children were playing with a ball the measured speed was 54.29 kph and the estimated speed 39.27 kph. When children were waiting to cross the measured speed was 52.78 kph, estimated speed 34.02 kph. Analyses indicated that there were significant differences between measured and estimated speeds. CONCLUSIONS: New Zealand drivers make inadequate speed adjustments in the presence of children, despite probably believing they do so. Establishing specific rules about appropriate speeds around children and highlighting to drivers the discrepancy between their attitudes and behaviour are two intervention strategies suggested. PMID- 12642558 TI - Injury prevention attitudes and awareness in New Zealand. AB - OBJECTIVES: This study was designed to obtain New Zealand data on beliefs related to a broad spectrum of injuries and their prevention. METHODS: A cross sectional phone survey was conducted of approximately 400 randomly selected households from each of 13 territorial local authorities across New Zealand, giving a total of 5282. Respondents were asked questions on awareness and attitudes to injury prevention, ownership and use of safety equipment, safety behaviours, and incidence of self reported injury. RESULTS: 84% agreed with the statement that "Most injuries are preventable" and 91% rated their homes as "very safe" or "reasonably safe". A high proportion of homes had smoke alarms (81%) and first aid kits (81%), and more than half (56%) had turned down the temperature of their hot water to 55 degrees C or lower. However, less than half of the respondents said that they practised the other safety behaviours. Significant associations were found between the practise of safety behaviours and respondents' home safety ratings. There was a significant association between home safety ratings and the incidence of injury occurring in all settings (p<0.0001), however there was no discernable association between home safety ratings and injury occurring in the home. CONCLUSIONS: Although this survey found that most respondents believed that injuries are preventable and considered their homes to be safe, the public need to be further encouraged to adopt common safety practices and behaviours in the home. PMID- 12642559 TI - Association between handgun purchase and mortality from firearm injury. AB - OBJECTIVE: To determine the association between mortality from violent or firearm related injury and previous handgun purchase. METHODS: Case-control study of 213 466 Californians ages 21 and older who died in 1998; cases were the 4728 violent or firearm related injury deaths, with subsets by specific cause and means of death, and controls were the 208 738 non-injury deaths. The exposure of interest was the purchase of a handgun during 1996-98. The main outcome measure was the odds ratio for handgun purchase, adjusted for age, sex, race, education, and marital status. RESULTS: Handgun purchase was more common among persons dying from suicide (odds ratio (OR) 6.8; 95% confidence interval (CI) 5.7 to 8.1) or homicide (OR 2.4, 95% CI 1.6 to 3.7), and particularly among those dying from gun suicide (OR 12.5; 95% CI 10.4 to 15.0) or gun homicide (OR 3.3; 95% CI 2.1 to 5.3), than among controls. No such differences were seen for non-gun suicide or homicide. Among women, those dying from gun suicide were much more likely than controls to have purchased a handgun (OR 109.8; 95% CI 61.6 to 195.7). Handgun purchasers accounted for less than 1% of the study population but 2.4% of gun homicides, 14.2% of gun suicides, and 16.7% of unintentional gun deaths. Gun suicide made up 18.9% of deaths among purchasers but only 0.6% of deaths among non-purchasers. CONCLUSION: Among adults who died in California in 1998, those dying from violence were more likely than those dying from non-injury causes to have purchased a handgun. PMID- 12642560 TI - Studying "exposure" to firearms: household ownership v access. AB - BACKGROUND: Firearm ownership has often been used to measure access to weapons. However, persons who own a firearm may not have access to it and conversely, persons who do not own a firearm may be able to access one quickly. OBJECTIVES: To examine whether using firearm ownership is a reasonable proxy for access by describing the demographic characteristics associated with ownership and access. METHODS: Data are from the 1994 Injury Control and Risk Survey, a national, random digit dial survey. Information about household firearm ownership and ready access to a loaded firearm were collected and weighted to provide national estimates. Adjusted odds ratios for three separate models were calculated using logistic regression. RESULTS: A total of 1353 (27.9%) respondents reported both having a firearm in the household and ready access to one. An additional 313 respondents (8.1%) reported having a firearm, but were not able to access these weapons. Another 421 respondents (7.2%) did not have a firearm in or around their home, yet reported being able to retrieve and fire one within 10 minutes. Based on the logistic regression findings, the demographic characteristics of this latter group are quite different from those who report ownership. Those who do not have a firearm, but report ready access to one, are more likely to be ethnic minorities, single, and living in attached homes. CONCLUSIONS: Asking only about the presence of a firearm in a household may miss some respondents with ready access to a loaded firearm. More importantly, those who do not own a firearm, but report ready access to one, appear to be qualitatively different from those who report ownership. Caution should be exercised when using measures of ownership as a proxy for access. PMID- 12642561 TI - Family characteristics and pedestrian injury risk in Mexican children. AB - BACKGROUND: Family characteristics have been described as risk factors for child pedestrian and motor vehicle collision. Research results come mainly from developed countries, where family relationships could be different than in developing ones. OBJECTIVE: To examine family characteristics as risk factors for pedestrian injury in children living in Guadalajara City, Mexico. METHODS: Case control study of injuries among children 1-14 years of age involved in pedestrian motor vehicle collisions. Cases resulting in death or injuries that required hospitalization or medical attention were included and identified through police reports and/or emergency room registries. Two neighborhood matched controls were selected randomly and compared with cases to estimate odds ratios (OR) and 95% confidence intervals (CI). RESULTS: Significant risk factors were: male (OR 2.3, 95% CI 1.2 to 4.4), number of siblings in household (two siblings, OR 3.2, 95% CI 1.4 to 6.6; three siblings, OR 4.5, 95% CI 1.9 to 11.0; four or more siblings, OR 3.7, 95% CI 1.1 to 12.9), and number of non-siblings/non-parents in household (four or more, OR 6.2, 95% CI 1.5 to 26.6). Children of a sole mother, working mother, or grandmother living in house did not show increased risk after adjusting for socioeconomic conditions. CONCLUSION: Household size has implications for child pedestrian and motor vehicle collision prevention efforts and is relatively easy to identify. Also, the lack of risk association with working mothers may indicate that grandmothers are not part of the social support network that cares for children of working mothers. PMID- 12642562 TI - Seismic, structural, and individual factors associated with earthquake related injury. AB - BACKGROUND: Earthquakes cause thousands of deaths worldwide every year, and systematic study of the causes of these deaths can lead to their prevention. Few studies have examined how multiple types of risk factors are related to physical injury during an earthquake. METHODS: A population based case-control study was conducted to examine how individual characteristics, building characteristics, and seismic features of the 1994 Northridge, California, earthquake contributed to physical injury. Cases included fatal and hospital-admitted injuries caused by the earthquake. Controls were drawn from a population based phone survey of county residents. Cases were individually matched to two sets of controls: one matched by age and gender and one matched by location at the time of the earthquake. RESULTS: Individuals over age 65 had 2.9 times the risk of injury as younger people (95% confidence interval (CI) 1.2 to 7.4) and women had a 2.4 times greater risk than men (95% CI 1.2 to 5.1). Location in multiple unit residential and commercial structures each led to increased injury risk compared with single unit residential structures, but the exact estimate varied depending on the control group used. With every increase in ground motion of 10%g, injury risk increased 2.2 times (95% CI 1.6 to 3.3). CONCLUSIONS: Controlling for other factors, it was found that individual, building, and seismic characteristics were independently predictive of increased injury risk. Prevention and preparedness efforts should focus on each of these as potential points of intervention. PMID- 12642563 TI - Suicide among North Carolina women, 1989-93: information from two data sources. AB - OBJECTIVES: To characterize the events and examine suicide precursors among women and to examine gaps in surveillance. SETTING: A statewide study in North Carolina. METHODS: Suicides of women age 15 and older for the time period 1989 93, as identified from the Office of the Chief Medical Examiner, were included. All case files were reviewed by hand and telephone interviews were attempted with investigating law enforcement officials for every case in 1993. RESULTS: Altogether 882 suicides met the case definition, for an age adjusted rate that fluctuated between 5.53 and 7.26 per 100 000 women across the period. Interviews with law enforcement officials were completed for 135 of the 177 cases from 1993. White women had rates nearly three times those of racial minorities. Women under age 45 were proportionally more likely than older women to have recently experienced the breakup of an intimate relationship. Information about precursors was not as consistently reported as had been hoped. Medical examiner records were variable in completeness. Law enforcement interviews frequently did not yield information about the factors we had hoped to examine, probably because the investigations were conducted primarily to rule out homicide. CONCLUSIONS: This study suggests somewhat different precursor patterns by age group. It also points to the need for reconsidering how suicide surveillance is accomplished as a strategy to guide intervention. PMID- 12642564 TI - Validity of self reported home safety practices. AB - OBJECTIVES: To examine the validity of self reported data on parents' home safety practices of using smoke alarms and stair gates, and having syrup of ipecac. SETTING: Families from a pediatric continuity clinic in a large, urban teaching hospital with infants from birth to 6 months were enrolled in the study. METHODS: As part of a randomized controlled trial to promote home safety, parents' responses to personal interviews were compared to observations made in the respondents' homes two to four weeks after the interview. Positive and negative predictive values, sensitivity, and specificity were computed and compared between the intervention and control group families. RESULTS: Sensitivities were high among the four safety practices. Specificities were much lower and fell into a much wider range than sensitivities. The positive predictive values were low and the negative predictive values were high. No differences in these indicators of validity were found between intervention and control group families. CONCLUSIONS: If the main interest in an evaluation is on the relative difference between study groups, rather than the absolute value of the outcome measure, our results suggest that self reported data may be of acceptable validity. However, when assessing a patient's risk, clinicians need to recognize the problem of over reporting of safety practices. PMID- 12642565 TI - Injuries and death of children in rollover motor vehicle crashes in the United States. AB - CONTEXT: There is an increased awareness of the problem of rollover crashes, but few data on children involved in rollover crashes in the United States. OBJECTIVE: To determine: (1) the rates of rollover crashes involving children and the incidence of fatal injury; (2) the characteristics of crashes involving children; (3) the risk factors for children being in a rollover compared with a non-rollover crash; and (4) whether the risk of death is greater for children involved in crashes in sport utility vehicles (SUVs) or passenger cars. DESIGN: Retrospective cohort study. DATA: 1993 through 1998 crashes involving children younger than 16 years included in the Crashworthiness Data System or reported to the Fatality Analysis Reporting System. RESULTS: During the study period, 100.4 children per 100 000 person-years were involved in a rollover crash, accounting for 10% of all children involved in crashes. The incidence of fatal injuries in rollover crashes was 3.4 per 100 000 person-years. Sixty percent of children involved in rollovers were riding in SUVs. Among vehicles carrying children and involved in a crash, the adjusted relative risk of the crash being a rollover was 11.1 (95% confidence interval (CI) 9.3 to 13.3) for SUVs compared with passenger cars. The adjusted relative risk of death was 1.8 (95% CI 1.1 to 2.8) in a rollover crash and the relative risk of injury was 2.1 (95% CI 1.1 to 3.8) compared with non-rollover crashes. However, the relative risk of death for children in SUVs which crashed was 0.4 (95% CI 0.1 to 2.5) compared with passenger cars which crashed. CONCLUSIONS: Crashes involving children in SUVs were more likely to be rollover crashes than those involving passenger cars, and rollover crashes were associated with an increased risk of death and injury. However, the overall risk of death for children in a crash was not higher for children who crashed in an SUV compared to children who crashed in a passenger vehicle. Whether children are safer overall in SUVs compared with other cars cannot be answered with the data used. PMID- 12642566 TI - Predictors and severity of injury in assaults with barglasses and bottles. AB - BACKGROUND: Although glasses and bottles are frequently used as weapons in assaults, there is little knowledge on which prevention strategies can be based. DESIGN: Scrutiny of a random sample of 1288 criminal injury compensation applications. OBJECTIVE: To identify predictors and relative severity of glass and bottle injury. METHOD: Injury site, severity, treatment, and demographic characteristics of victims and assailants were studied with reference to awards from the UK national Criminal Injuries Compensation Authority (CICA). MAIN OUTCOME MEASURES: Gender of victims and assailants, injury sites, treatment, and award (UK pounds) as indices of injury severity. RESULTS: Annual CICA awards to all victims of assaults in licensed premises during 1996-98 amounted to pound 4.08 million (for all glass/bottle assaults: pound 1.15 million = 28%). The mean cost of 746 glass assaults was pound 2347, compared with pound 2007 for 542 injuries from bottle assaults (mean difference pound 340; p<0.01). This difference largely reflected more eye injuries with glasses (26 cases: 3% of all glass assaults) than with bottles (eight cases: 1% of all bottle assaults). Bottle assault was significantly associated with unidentified assailants and scalp injuries; whereas glass injury was significantly linked to pub opening hours (midday to midnight), Thursdays, eye and face injuries, and treatment requiring sutures. Mean age of bottle assault victims (26.1 years) was lower than of glass victims (27.3 years; p<0.01), and same gender assaults were more frequent than between gender assaults for both bottle (p<0.001) and glass (p<0.001) assaults. Female victims were allocated to lower compensation awards more frequently than male victims; this was the case for both bottle (p<0.05) and glass (p<0.01) assaults. CONCLUSIONS: Assaults with bottles caused less serious injury and resulted in lower compensation costs. Injury distribution was linked to victim gender and weapon choice, but not to assailant gender. Prevention strategies should focus on both bottle and glass assaults and should take account of the setting and time in which drinking occurs. PMID- 12642568 TI - Children's response to a commercial back-up warning device. AB - OBJECTIVE: To determine preschool children's response to a commercial back-up warning alarm in a mock setting of an automobile backing up. SETTING: Preschool parking lot in Albuquerque, New Mexico, USA. METHODS: With subjects acting as their own controls, 33 preschoolers were asked to walk behind a stationary vehicle twice. The first time, the control, no warning sound was emitted from the vehicle. The second time, the vehicle was placed in reverse gear triggering an alarm. Children's responses were recorded by a hidden video camera. Avoidance behavior by the child was considered a positive response. Location and distance to where the response occurred was measured. RESULTS: Thirty three children, 38 61 months, were studied. None responded to the alarm with avoidance behavior but 18 looked toward the vehicle or hesitated in their gait. CONCLUSION: Although over half of the children acknowledged the warning alarm, the device did not elicit avoidance behavior. Mere acknowledgment of the warning device would not prevent injury. PMID- 12642567 TI - An investigation of child restraint/seatbelt usage in motor vehicles by Maori in Northland New Zealand. AB - OBJECTIVE: To investigate child restraint/seatbelt use by the indigenous (Maori) population in Northland New Zealand. METHOD: Observational surveys were conducted at the two main car parks (McDonald's and the largest supermarket) to determine the number of passengers restrained, the type of restraints, and correct use. Observations were restricted to those who were obviously Maori, based upon the local knowledge of the observer. In addition, face to face questionnaires were administered to Maori whanau/caregivers involved in the care of two or more children for more than three days a week. RESULTS: A total of 788 participants were observed. Babies were those most likely to have all occupants restrained correctly (97%), followed by toddlers (66%), adults (56%), and school age children (48%); 138 interviews were conducted. Females (86%) were significantly more likely to ensure that all passengers were restrained on short journeys compared to males (67%; p<0.05). Respondents under 45 (80%) were significantly less likely to restrain child passengers compared to people aged 45 or older (91%; p<0.05). DISCUSSION: This study highlights the problem that larger families in this study had in providing correct child restraints for all their children. PMID- 12642569 TI - How much science is there in injury prevention and control? AB - OBJECTIVES: To determine what proportion of research papers at an injury prevention conference reported an evaluation. METHODS: A random sample of 250 abstracts from the 6th World Conference on Injury Prevention and Control were classified by methodological type. Those that described any evaluation were further subdivided by whether the evaluation was of process or if it used an intermediate or "true" outcome. RESULTS: Of 250 abstracts, 20 (8%; 95% confidence interval 5.0% to 12.1%) showed evaluations with intermediate or true outcomes. Research designs were weak. Among the 20 reports, none was a randomized trial and only two conducted a before and after study with control group. The remaining 17 used before-after or "after only" designs. CONCLUSION: The conference papers included few evaluations. To ensure that resources are best used, those in the injury prevention field must increase their use of rigorous evaluation. PMID- 12642570 TI - Bias in estimates of seat belt effectiveness. PMID- 12642571 TI - Misclassification of seat belt use. PMID- 12642572 TI - No tea until three? PMID- 12642574 TI - Reconstitution of membrane proteolysis by FtsH. AB - Escherichia coli FtsH is a membrane-bound and ATP-dependent protease responsible for degradation of several membrane proteins. The FtsH action is processive and presumably involves dislocation of the substrate from the membrane to the cytosol. Although elucidation of its molecular mechanism requires an in vitro reaction system, in vitro activities of this enzyme against membrane protein substrates have only been assayed using detergent-solubilized components. Here we report on the construction of in vitro reaction systems for FtsH-catalyzed membrane protein degradation. A combination of two inverted membrane vesicles or of two proteoliposomes, one bearing the enzyme and the other bearing a substrate, was fused by polyethylene glycol 3350 treatment. Addition of ATP then resulted in degradation of the substrate. It was shown that FtsH can function in the process of membrane proteins degradation without aid from any other cellular factors. PMID- 12642575 TI - High resolution x-ray structure of tyvelose epimerase from Salmonella typhi. AB - Tyvelose epimerase catalyzes the last step in the biosynthesis of tyvelose by converting CDP-d-paratose to CDP-d-tyvelose. This unusual 3,6-dideoxyhexose occurs in the O-antigens of some types of Gram-negative bacteria. Here we describe the cloning, protein purification, and high-resolution x-ray crystallographic analysis of tyvelose epimerase from Salmonella typhi complexed with CDP. The enzyme from S. typhi is a homotetramer with each subunit containing 339 amino acid residues and a tightly bound NAD+ cofactor. The quaternary structure of the enzyme displays 222 symmetry and can be aptly described as a dimer of dimers. Each subunit folds into two distinct lobes: the N-terminal motif responsible for NAD+ binding and the C-terminal region that harbors the binding site for CDP. The analysis described here demonstrates that tyvelose epimerase belongs to the short-chain dehydrogenase/reductase superfamily of enzymes. Indeed, its active site is reminiscent to that observed for UDP-galactose 4 epimerase, an enzyme that plays a key role in galactose metabolism. Unlike UDP galactose 4-epimerase where the conversion of configuration occurs about C-4 of the UDP-glucose or UDP-galactose substrates, in the reaction catalyzed by tyvelose epimerase, the inversion of stereochemistry occurs at C-2. On the basis of the observed binding mode for CDP, it is possible to predict the manner in which the substrate, CDP-paratose, and the product, CDP-tyvelose, might be accommodated within the active site of tyvelose epimerase. PMID- 12642577 TI - The G-protein regulator AGS3 controls an early event during macroautophagy in human intestinal HT-29 cells. AB - AGS3 contains GoLoco or G-protein regulatory motifs in its COOH-terminal half that stabilize the GDP-bound conformation of the alpha-subunit of the trimeric Gi3 protein. The latter is part of a signaling pathway that controls the lysosomal-autophagic catabolism in human colon cancer HT-29 cells. In the present work we show that the mRNA encoding for AGS3 is expressed in human intestinal cell lines (Caco-2 and HT-29) whatever their state of differentiation. Together with the full-length form, minute amounts of the mRNA encoding a NH2-terminal truncated form of AGS3, previously characterized in cardiac tissues, were also detected. Both the endogenous form of AGS3 and a tagged expressed form have a localization compatible with a role in the Galphai3-dependent control of autophagy. Accordingly, expressing its non-Galphai3-interacting NH2-terminal domain or its Galphai3-interacting COOH-terminal domain reversed the stimulatory role of AGS3 on autophagy. On the basis of biochemical and morphometric analysis, we conclude that AGS3 is involved in an early event during the autophagic pathway probably prior to the formation of the autophagosome. These data demonstrate that AGS3 is a novel partner of the Galphai3 protein in the control of a major catabolic pathway. PMID- 12642576 TI - Autotaxin is released from adipocytes, catalyzes lysophosphatidic acid synthesis, and activates preadipocyte proliferation. Up-regulated expression with adipocyte differentiation and obesity. AB - Our group has recently demonstrated (Gesta, S., Simon, M., Rey, A., Sibrac, D., Girard, A., Lafontan, M., Valet, P., and Saulnier-Blache, J. S. (2002) J. Lipid Res. 43, 904-910) the presence, in adipocyte conditioned-medium, of a soluble lysophospholipase d-activity (LPLDact) involved in synthesis of the bioactive phospholipid lysophosphatidic acid (LPA). In the present report, LPLDact was purified from 3T3F442A adipocyte-conditioned medium and identified as the type II ecto-nucleotide pyrophosphatase phosphodiesterase, autotaxin (ATX). A unique ATX cDNA was cloned from 3T3F442A adipocytes, and its recombinant expression in COS-7 cells led to extracellular release of LPLDact. ATX mRNA expression was highly up regulated during adipocyte differentiation of 3T3F442A-preadipocytes. This up regulation was paralleled by the ability of newly differentiated adipocytes to release LPLDact and LPA. Differentiation-dependent up-regulation of ATX expression was also observed in a primary culture of mouse preadipocytes. Treatment of 3T3F442A-preadipocytes with concentrated conditioned medium from ATX expressing COS-7 cells led to an increase in cell number as compared with concentrated conditioned medium from ATX non-expressing COS-7 cells. The specific effect of ATX on preadipocyte proliferation was completely suppressed by co treatment with a LPA-hydrolyzing phospholipase, phospholipase B. Finally, ATX expression was found in mature adipocytes isolated from mouse adipose tissue and was substantially increased in genetically obese-diabetic db/db mice when compared with their lean siblings. In conclusion, the present work shows that ATX is responsible for the LPLDact released by adipocytes and exerts a paracrine control on preadipocyte growth via an LPA-dependent mechanism. Up-regulations of ATX expression with adipocyte differentiation and genetic obesity suggest a possible involvement of this released protein in the development of adipose tissue and obesity-associated pathologies. PMID- 12642578 TI - Opioid agonists have different efficacy profiles for G protein activation, rapid desensitization, and endocytosis of mu-opioid receptors. AB - The differential ability of various mu-opioid receptor (MOP) agonists to induce rapid receptor desensitization and endocytosis of MOP could arise simply from differences in their efficacy to activate G proteins or, alternatively, be due to differential capacity for activation of other signaling processes. We used AtT20 cells stably expressing a low density of FLAG-tagged MOP to compare the efficacies of a range of agonists to 1) activate G proteins using inhibition of calcium channel currents (ICa) as a reporter before and after inactivation of a fraction of receptors by beta-chlornaltrexamine, 2) produce rapid, homologous desensitization of ICa inhibition, and 3) internalize receptors. Relative efficacies determined for G protein coupling were [Tyr-D-Ala-Gly-MePhe Glyol]enkephalin (DAMGO) (1) > or = methadone (0.98) > morphine (0.58) > pentazocine (0.15). The same rank order of efficacies for rapid desensitization of MOP was observed, but greater concentrations of agonist were required than for G protein activation. By contrast, relative efficacies for promoting endocytosis of MOP were DAMGO (1) > methadone (0.59) >> morphine (0.07) > or = pentazocine (0.03). These results indicate that the efficacy of opioids to produce activation of G proteins and rapid desensitization is distinct from their capacity to internalize mu-opioid receptors but that, contrary to some previous reports, morphine can produce rapid, homologous desensitization of MOP. PMID- 12642579 TI - Structural determinants of the regulation of the voltage-gated potassium channel Kv2.1 by the modulatory alpha-subunit Kv9.3. AB - Voltage-gated potassium (Kv) channels containing alpha-subunits of the Kv2 subfamily mediate delayed rectifier currents in excitable cells. Channels formed by Kv2.1 alpha-subunits inactivate from open- and closed states with both forms of inactivation serving different physiological functions. Here we show that open and closed-state inactivation of Kv2.1 can be distinguished by the sensitivity to intracellular tetraethylammonium and extracellular potassium and lead to the same inactivated conformation. The functional properties of Kv2.1 are regulated by its association with modulatory alpha-subunits (Kv5, Kv6, Kv8, and Kv9). For instance, Kv9.3 changes the state preference of Kv2.1 inactivation by accelerating closed-state inactivation and inhibiting open-state inactivation. An N-terminal regulatory domain (NRD) has been suggested to determine the function of the modulatory alpha-subunit Kv8.1. However, when we tested the NRD of Kv9.3, we found that the functional properties of chimeric Kv2.1 channels containing the NRD of Kv9.3 (Kv2.1(NRD)) did not resemble those of Kv2.1/Kv9.3 heteromers, thus questioning the role of the NRD in Kv9 subunits. A further region of interest is a PXP motif in the sixth transmembrane segment. This motif is conserved among all alpha-subunits of the Kv1, Kv2, Kv3, and Kv4 subfamilies, whereas the second proline is not conserved in any modulatory alpha-subunit. Exchanging this proline in Kv2.1 for the corresponding residue of Kv9.3 resulted in channels (Kv2.1 P410T) that show all hallmarks of the regulation of Kv2.1 by Kv9.3. The effect prevailed in heteromeric channels following co-expression of Kv2.1-P410T with Kv2.1. These data suggest that the alteration of the PXP motif is an important determinant of the regulatory function of modulatory alpha-subunits. PMID- 12642580 TI - Roles of Src and epidermal growth factor receptor transactivation in transient and sustained ERK1/2 responses to gonadotropin-releasing hormone receptor activation. AB - The duration as well as the magnitude of mitogen-activated protein kinase activation has been proposed to regulate gene expression and other specific intracellular responses in individual cell types. Activation of ERK1/2 by the hypothalamic neuropeptide gonadotropin-releasing hormone (GnRH) is relatively sustained in alpha T3-1 pituitary gonadotropes and HEK293 cells but is transient in immortalized GT1-7 neurons. Each of these cell types expresses the epidermal growth factor receptor (EGFR) and responds to EGF stimulation with significant but transient ERK1/2 phosphorylation. However, GnRH-induced ERK1/2 phosphorylation caused by EGFR transactivation was confined to GT1-7 cells and was attenuated by EGFR kinase inhibition. Neither EGF nor GnRH receptor activation caused translocation of phospho-ERK1/2 into the nucleus in GT1-7 cells. In contrast, agonist stimulation of GnRH receptors expressed in HEK293 cells caused sustained phosphorylation and nuclear translocation of ERK1/2 by a protein kinase C-dependent but EGFR-independent pathway. GnRH-induced activation of ERK1/2 was attenuated by the selective Src kinase inhibitor PP2 and the negative regulatory C-terminal Src kinase in GT1-7 cells but not in HEK293 cells. In GT1-7 cells, GnRH stimulated phosphorylation and nuclear translocation of the ERK1/2-dependent protein, p90RSK-1 (RSK-1). These results indicate that the duration of ERK1/2 activation depends on the signaling pathways utilized by GnRH in specific target cells. Whereas activation of the Gq/protein kinase C pathway in HEK293 cells causes sustained phosphorylation and translocation of ERK1/2 to the nucleus, transactivation of the EGFR by GnRH in GT1-7 cells elicits transient ERK1/2 signals without nuclear accumulation. These findings suggest that transactivation of the tightly regulated EGFR can account for the transient ERK1/2 responses that are elicited by stimulation of certain G protein-coupled receptors. PMID- 12642581 TI - Pituitary tumor AP-2alpha recognizes a cryptic promoter in intron 4 of fibroblast growth factor receptor 4. AB - Fibroblast growth factor receptors (FGFRs) have been implicated in a multitude of proliferative functions, and FGFR4 is expressed differentially in normal and neoplastic pituitary. Human pituitary tumors express a truncated FGFR4 isoform (ptd-FGFR4) for which transcription is initiated from a downstream alternative site. Analysis of FGFR4 intronic sequences predicted a possible promoter within intron 4 (In4) including a classic TATA box with a possible transcriptional start site in intron 5. We show here that the human In4 sequence can direct luciferase reporter activity in transfected pituitary GH4 cells. Four overlapping fragments (A1, A2, B1, and B2) of this intron were examined by electromobility shift assay using nuclear extracts from rat pituitary tumors. Of these, fragment B2 formed complexes with nuclear rat pituitary GH4 extracts that were competed specifically by wild type but not mutant oligonucleotides for the neural crest cell lineage derived activating transcription factor AP-2. Conversely, an AP-2 consensus sequence probe was competed by the In4 B2 oligonucleotide but not by other fragments of the same intron. The In4 B2 complex was competed partially by NFkappaB, supershifted by an AP-2alpha-specific antibody, and co-migrated with the same probe incubated with recombinant AP-2alpha protein. We also examined the ability of primary human pituitary tumor extracts to interact with the In4 B2 fragment. Pituitary tumor-In4 B2 complexes were competed specifically by wild type AP-2 but not mutant AP-2 oligonucleotides. Western blotting revealed higher levels of AP-2alpha expression in primary human pituitary tumors than in nontumorous tissue. Mutagenesis of the putative AP-2 binding site in In4 B2 resulted in a marked loss of promoter activity in a luciferase assay. AP-2alpha transfection in the presence of the histone deacetylase inhibitor trichostatin-A resulted in enhanced expression of endogenous ptd-FGFR4. These data indicate that a cryptic promoter within intron 4 binds AP-2alpha. AP-2alpha and chromatin changes may contribute to the utilization of an alternative transcription start site leading to the genesis of the tumorigenic ptd-FGFR4 isoform. PMID- 12642582 TI - Erythroid gene suppression by NF-kappa B. AB - NF-kappa B/Rel transcription factors play essential roles to mediate the immune response and apoptosis, and they have also been implicated in cellular differentiation such as erythropoiesis. To elucidate the possible role(s) of NF kappa B in erythroid gene regulation and erythropoiesis, we have carried out transient transfection studies of the human embryonic/fetal erythroid cell line K562 and mouse adult erythroid MEL cells. It is shown that tumor necrosis factor alpha represses the transcription activity directed by either alpha or zeta globin promoter in a dose-dependent manner. Furthermore, different NF-kappa B family members could effectively repress the transfected alpha-like globin promoters in K562 as well as in MEL cells. The involvement of NF-kappa B pathway is supported by the ability of a NF-kappa B-specific, dominant negative mutant to block the tumor necrosis factor-alpha or p65-mediated suppression of the alpha like globin promoter activities. The suppression appears to be mediated through cis-linked HS-40 enhancer. Finally, stably transfected K562 cells overexpressing p65 contain reduced amounts of the p45/NF-E2 RNA and functional NF-E2 proteins. Our studies have identified a new set of targets of NF-kappa B. We suggest that the relatively high activity of the NF-kappa B pathway in early erythroid progenitors is involved in the suppression of erythroid-specific genes. Later in differentiation, together with other changes, the decline of the amounts of the NF-kappa B family of factors leads to derepression and consequent increase of NF E2, which in turn would activate a subset of erythroid-specific genes. PMID- 12642583 TI - Poly(ADP-ribose) polymerase-1 is a positive regulator of the p53-mediated G1 arrest response following ionizing radiation. AB - Poly(ADP-ribose) polymerase-1 (PARP-1) and the p53 tumor suppressor protein are both involved in the cellular response to genotoxic stress. Upon binding to the site of DNA strand breakage, PARP-1 is activated, leading to rapid and transient poly(ADP-ribosyl)ation of nuclear proteins using NAD+ as substrate. To investigate the role of PARP-1 in the p53 response to ionizing radiation in human cells, PARP-1 function was disrupted in wild-type p53 expressing MCF-7 and BJ/TERT cells using two strategies: chemical inhibition with 1,5 dihydroxyisoquinoline, and trans-dominant inhibition by overexpression of the PARP-1 DNA-binding domain. Although a number of proteins can catalyze poly(ADP ribosyl)ation in addition to PARP-1, we show that PARP-1 is the only detectable active species in BJ/TERT and MCF-7 cells. 1,5-Dihydroxyisoquinoline treatment prior to ionizing radiation delayed and attenuated the induction of two p53 responsive genes, p21 and mdm-2, and led to suppression of the p53-mediated G1 arrest response in MCF-7 and BJ/TERT cells. Trans-dominant inhibition of PARP-1 by overexpression of the PARP-1 DNA-binding domain in MCF-7 cells also led to a delay and attenuation in p21 induction and suppression of the p53-mediated G1 arrest response to ionizing radiation. Hence, inhibition of endogenous PARP-1 function suppresses the transactivation function of p53 in response to ionizing radiation. This study establishes PARP-1 as a critical regulator of the p53 response to DNA damage. PMID- 12642584 TI - Allosteric modulation of human P-glycoprotein. Inhibition of transport by preventing substrate translocation and dissociation. AB - The human multidrug transporter P-glycoprotein (Pgp, ABCB1) contributes to the poor bioavailability of many anticancer and antimicrobial agents as well as to drug resistance at the cellular level. For rational design of effective Pgp inhibitors, a clear understanding of its mechanism of action and functional regulation is essential. In this study, we demonstrate that inhibition of Pgp mediated drug transport by cis-(Z)-flupentixol, a thioxanthene derivative, occurs through an allosteric mechanism. Unlike competitive inhibitors, such as cyclosporin A and verapamil, cis-(Z)-flupentixol does not interfere with substrate ([(125)I]iodoarylazidoprazosin) recognition by Pgp, instead it prevents substrate translocation and dissociation, resulting in a stable but reversible Pgp-substrate complex. cis-(Z)-Flupentixol-induced complex formation requires involvement of the Pgp substrate site, because agents that either physically compete (cyclosporin A) for or indirectly occlude (vanadate) the substrate binding site prevent formation of the complex. Allosteric modulation by cis-(Z) flupentixol involves a conformational change in Pgp detectable by monoclonal antibody UIC2 binding to a conformation-sensitive external epitope of Pgp. The conformational change observed is distinct from that induced by Pgp substrates or competitive inhibitors. A single amino acid substitution (F983A) in TM12 of Pgp that impairs inhibition by cis-(Z)-flupentixol of Pgp-mediated drug transport also affects stabilization of the Pgp-substrate complex as well as the characteristic conformational change. Taken together, our results describe the molecular mechanism by which the Pgp modulator cis-(Z)-flupentixol allosterically inhibits drug transport. PMID- 12642585 TI - Mitochondrial functional state in clonal pancreatic beta-cells exposed to free fatty acids. AB - Excessive free fatty acid (FFA) exposure represents a potentially important diabetogenic condition that can impair insulin secretion from pancreatic beta cells. Because mitochondrial oxidative phosphorylation is a main link between glucose metabolism and insulin secretion, in the present work we investigated the effects of the FFA oleate (OE) on mitochondrial function in the clonal pancreatic beta-cell line, MIN6. Both the long term (72 h) and short term (immediately after application) impact of OE exposure on beta-cells was investigated. After 72 h of exposure to OE (0.4 mm, 0.5% bovine serum albumin) cells were washed and permeabilized, and mitochondrial function (respiration, phosphorylation, membrane potential formation, production of reactive oxygen species) was measured in the absence or presence of OE. MIN6 cells exposed to OE for 72 h showed impaired glucose-stimulated insulin secretion and decreased cellular ATP. Mitochondria in OE-exposed cells retained normal functional characteristics in FFA-free medium; however, they were significantly more sensitive to the acute uncoupling effect of OE treatment. The mitochondria of OE-exposed cells displayed increased depolarization caused by acute OE treatment, which is attributable to the elevation in the FFA-transporting function of uncoupling protein 2 and the dicarboxylate carrier. These cells also had an increased production of reactive oxygen species in complex I of the mitochondrial respiratory chain that could be activated by FFA. A high level of reduction of respiratory complex I augmented acute FFA-induced uncoupling in a way compatible with activation of mitochondrial uncoupling protein by intramitochondrial superoxide. A stronger augmentation was observed in OE-exposed cells. Together, these events may underlie FFA-induced depression of the ATP/ADP ratio in beta-cells, which accounts for the defective glucose-stimulated insulin secretion associated with lipotoxicity. PMID- 12642586 TI - Minimal BH3 peptides promote cell death by antagonizing anti-apoptotic proteins. AB - The pro-apoptotic "BH3 domain-only" proteins of the Bcl-2 family (e.g. Bid and Bad) transduce multiple death signals to the mitochondrion. They interact with the anti-apoptotic Bcl-2 family members and induce apoptosis by a mechanism that requires the presence of at least one of the multidomain pro-apoptotic proteins Bax or Bak. Although the BH3 domain of Bid can promote the pro-apoptotic assembly and function of Bax/Bak by itself, other BH3 domains do not function as such. The latter point raises the question of whether, and how, these BH3 domains induce apoptosis. We show here that a peptide comprising the minimal BH3 domain from Bax induces apoptosis but is unable to stimulate the apoptotic activity of microinjected recombinant Bax. This relies on the inability of the peptide to directly induce Bax translocation to mitochondria or a change in its conformation. This peptide nevertheless interferes with Bax/Bcl-xL interactions in vitro and stimulates the apoptotic activity of Bax when combined with Bcl-xL. Similarly, a peptide derived from the BH3 domain of Bad stimulates Bax activity only in the presence of Bcl-xL. Thus, BH3 domains do not necessarily activate multidomain pro-apoptotic proteins directly but promote apoptosis by releasing active multidomain pro-apoptotic proteins from their anti-apoptotic counterparts. PMID- 12642587 TI - Induction of plasminogen activator inhibitor-1 by urokinase in lung epithelial cells. AB - The plasminogen/plasmin system, urokinase-type plasminogen activator (uPA), its receptor (uPAR), and its inhibitor (PAI-1), influence extracellular proteolysis and cell migration in lung injury or neoplasia. In this study, we sought to determine whether tcuPA (two chain uPA) alters expression of its major inhibitor PAI-1 in lung epithelial cells. The expression of PAI-1 was evaluated at the protein and mRNA level by Western blot, immunoprecipitation, and Northern blot analyses. We found that tcuPA treatment enhanced PAI-1 protein and mRNA expression in Beas2B lung epithelial cells in a time- and concentration-dependent manner. The tcuPA-mediated induction of PAI-1 involves post-transcriptional control involving stabilization of PAI-1 mRNA. Inactivation of the catalytic activity of tcuPA had little effect on PAI-1 induction and the activity of the isolated amino-terminal fragment was comparable with full-length single- or two chain uPA. In contrast, deletion of either the uPA receptor binding growth factor domain or kringle domain (kringle) from full-length single chain uPA markedly attenuated the induction of PAI-1. Induction of PAI-1 by exposure of lung epithelial cells to uPA is a newly recognized pathway by which PAI-1 could regulate local fibrinolysis and urokinase-dependent cellular responses in the setting of lung inflammation or neoplasia. PMID- 12642588 TI - Identification of Ets-1 as an important transcriptional activator of CTP:phosphocholine cytidylyltransferase alpha in COS-7 cells and co-activation with transcriptional enhancer factor-4. AB - Phosphatidylcholine biosynthesis via the CDP-choline pathway is primarily regulated by CTP:phosphocholine cytidylyltransferase (CT). Transcriptional enhancer factor-4 (TEF-4) enhances the transcription of CTalpha in COS-7 cells by interactions with the basal transcription machinery (Sugimoto, H., Bakovic, M., Yamashita, S., and Vance, D.E. (2001) J. Biol. Chem. 276,12338-12344). To identify the most important transcription factor involved in basal CTalpha transcription, we made CTalpha promoter-deletion and -mutated constructs linked to a luciferase reporter and transfected them into COS-7 cells. The results indicate that an important site regulating basal CTalpha transcription is -53/-47 (GACTTCC), which is a putative consensus-binding site of Ets transcription factors (GGAA) in the opposite orientation. Gel shift analyses indicated the existence of a binding protein for -53/-47 (GACTTCC) in nuclear extracts of COS-7 cells. When anti-Ets-1 antibody was incubated with the probe in gel shift analyses, the intensity of the binding protein was decreased. The binding of endogenous Ets-1 to the promoter probe was increased when TEF-4 was expressed; however, the amount of Ets-1 detected by immunoblotting was unchanged. When cells were transfected with Ets-1 cDNA, the luciferase activity of CTalpha promoter constructs was greatly enhanced. Co-transfection experiments with Ets-1 and TEF-4 showed enhanced expression of reporter constructs as well as CTalpha mRNA. These results suggest that Ets-1 is an important transcriptional activator of the CTalpha gene and that Ets-1 activity is enhanced by TEF-4. PMID- 12642589 TI - Induction of heme oxygenase-1 expression in murine macrophages is essential for the anti-inflammatory effect of low dose 15-deoxy-Delta 12,14-prostaglandin J2. AB - 15-Deoxy-Delta 12,14-prostaglandin J2 (15d-PGJ2), a cyclopentenone prostaglandin, displays a potent anti-inflammatory effect at micromolar concentrations (>2 microM) through direct inhibition of nuclear factor (NF)-kappa B activation. Here we show that at submicromolar concentrations (0.1-0.5 microM) 15d-PGJ2 retains the ability to suppress the production of tumor necrosis factor-alpha (TNF-alpha) and nitric oxide (NO) in lipopolysaccharide (LPS)-activated murine J774 macrophages under the conditions of a prolonged incubation (>12 h). Western blot analysis revealed that the expression of the cytoprotective enzyme, heme oxygenase-1 (HO-1), was induced and coincident with the anti-inflammatory action of 15d-PGJ2. Inhibition of HO-1 activity or scavenging carbon monoxide (CO), a byproduct derived from heme degradation, significantly attenuated the suppressive activity of 15d-PGJ2. Furthermore, LPS-induced NF-kappa B activation assessed by the inhibitory protein of NF-kappa B(I kappa B) degradation and p50 nuclear translocation was diminished in cells subjected to prolonged treatment with the low concentration of 15d-PGJ2. Treatment of cells with the protein synthesis inhibitor, cycloheximide, or the specific p38 MAP kinase inhibitor, SB203580, blocked the induction of HO-1 and suppression of LPS-induced I kappa B degradation mediated by 15d-PGJ2. Likewise, HO inhibitor and CO scavenger were effective in abolishing the inhibitory effects of 15d-PGJ2 on NF-kappa B activation induced by LPS. The functional role of CO was further demonstrated by the use of a CO releasing molecule, tricarbonyldichlororuthenium(II) dimer, which significantly suppressed LPS-induced nuclear translocation of p50 as assessed by confocal immunofluorescence. Collectively, these data suggest that even at submicromolar concentrations 15d-PGJ2 can exert an anti-inflammatory effect in macrophages through a mechanism that involves the action of HO/CO. PMID- 12642590 TI - Dynamic changes of fibrin architecture during fibrin formation and intrinsic fibrinolysis of fibrin-rich clots. AB - Clotting and fibrinolysis are initiated simultaneously in vivo, and fibrinolysis usually occurs without any individualized lysis front (intrinsic fibrinolysis). We have developed a novel model to assess whether morphological changes resulting from intrinsic fibrinolysis are similar to those previously reported at the lysis front using externally applied lytic agents. Fibrin assembly and fibrinolysis were followed in real-time by confocal microscopy using gold-labeled fibrinogen molecules. An increase in fiber absorbance (30%, p < 0.01) and a decrease in fiber diameter (60%, p < 0.01) due to the ongoing accumulation and packing of fibrin molecules were the most significant detectable features occurring during fibrin assembly. Similar features with a similar magnitude were observed during fibrin dissolution, but in the reverse order and with a 3-fold increase in duration. Then, lysing fibers were progressively transected laterally, and thinner fibers were cleaved at a 2.5-fold faster rate than thicker fibers (p < 0.001). Frayed lysing fibers were seen to interact progressively with adjoining fibers (agglomeration), leading to a 76 and 88% increase in the network pore diameter (p < 0.05) and fiber diameter (p < 0.01), respectively. At the maximum decrease in fiber absorbance (46%, p < 0.05), the network suddenly collapsed with the release of large fragments that gradually vanished. Morphological changes of fibrin that occur during intrinsic fibrinolysis are similar as those observed next to the lysis front, although they are not restricted spatially to the clot/surrounding milieu interface but are observed through the entire clot. PMID- 12642591 TI - Selective hydrolysis of triple-helical substrates by matrix metalloproteinase-2 and -9. AB - The role of proteases in the tumor cell invasion process is multifaceted. Members of the matrix metalloproteinase (MMP) family have been implicated in primary and metastatic tumor growth, angiogenesis, and degradation of extracellular matrix (ECM) components. Differentiating between the up-regulation of MMP production and the presence of activated MMPs can be difficult but may well dictate which MMPs are critical to invasion. Because the hydrolysis of collagens is one of the committed steps in ECM turnover, we have investigated selective MMP action on collagenous substrates as a means to evaluate active MMPs. Two triple-helical peptide (THP) models of the MMP-9 cleavage site in type V collagen, alpha1(V)436 450 THP and alpha1(V)436-447 fTHP, were hydrolyzed by MMP-2 and MMP-9 at the Gly Val bond, analogous to the bond cleaved by MMP-9 in the corresponding native collagen. Kinetic analyses showed k(cat)/K(m) values of 14,002 and 5,449 s(-1)m( 1) for MMP-2 and -9 hydrolysis of alpha1(V)436-447 fTHP, respectively. These values, along with individual k(cat) and K(m) values, are comparable with collagen hydrolysis by MMP-2 and -9. Neither THP was hydrolyzed by MMP-1, -3, 13, or -14. alpha1(V)436-447 fTHP and a general fluorogenic THP were used to screen for triple-helical peptidase activity in alpha(2)beta(1) integrin stimulated melanoma cells. Binding of the alpha(2)beta(1) integrin resulted in the production of substantial triple-helical peptidase activity, the majority (>95%) of which was non-MMP-2/-9. THPs were found to provide highly selective substrates for members of the MMP family and can be used to evaluate active MMP production in cellular systems. PMID- 12642592 TI - Palmitoylation regulates regulator of G-protein signaling (RGS) 16 function. II. Palmitoylation of a cysteine residue in the RGS box is critical for RGS16 GTPase accelerating activity and regulation of Gi-coupled signalling. AB - Palmitoylation is a reversible post-translational modification used by cells to regulate protein activity. The regulator of G-protein signaling (RGS) proteins RGS4 and RGS16 share conserved cysteine (Cys) residues that undergo palmitoylation. In the accompanying article (Hiol, A., Davey, P. C., Osterhout, J. L., Waheed, A. A., Fischer, E. R., Chen, C. K., Milligan, G., Druey, K. M., and Jones, T. L. Z. (2003) J. Biol. Chem. 278, 19301-19308), we determined that mutation of NH2-terminal cysteine residues in RGS16 (Cys-2 and Cys-12) reduced GTPase accelerating (GAP) activity toward a 5-hydroxytryptamine (5-HT1A)/G alpha o1 receptor fusion protein in cell membranes. NH2-terminal acylation also permitted palmitoylation of a cysteine residue in the RGS box of RGS16 (Cys-98). Here we investigated the role of internal palmitoylation in RGS16 localization and GAP activity. Mutation of RGS16 Cys-98 or RGS4 Cys-95 to alanine reduced GAP activity on the 5-HT1A/G alpha o1 fusion protein and regulation of adenylyl cyclase inhibition. The C98A mutation had no effect on RGS16 localization or GAP activity toward purified G-protein alpha subunits. Enzymatic palmitoylation of RGS16 resulted in internal palmitoylation on residue Cys-98. Palmitoylated RGS16 or RGS4 WT but not C98A or C95A preincubated with membranes expressing 5-HT1a/G alpha o1 displayed increased GAP activity over time. These results suggest that palmitoylation of a Cys residue in the RGS box is critical for RGS16 and RGS4 GAP activity and their ability to regulate Gi-coupled signaling in mammalian cells. PMID- 12642593 TI - Palmitoylation regulates regulators of G-protein signaling (RGS) 16 function. I. Mutation of amino-terminal cysteine residues on RGS16 prevents its targeting to lipid rafts and palmitoylation of an internal cysteine residue. AB - Regulators of G-protein signaling (RGS) proteins down-regulate signaling by heterotrimeric G-proteins by accelerating GTP hydrolysis on the G alpha subunits. Palmitoylation, the reversible addition of palmitate to cysteine residues, occurs on several RGS proteins and is critical for their activity. For RGS16, mutation of Cys-2 and Cys-12 blocks its incorporation of [3H]palmitate and ability to turn off Gi and Gq signaling and significantly inhibited its GTPase activating protein activity toward aG alpha subunit fused to the 5-hydroxytryptamine receptor 1A, but did not reduce its plasma membrane localization based on cell fractionation studies and immunoelectron microscopy. Palmitoylation can target proteins, including many signaling proteins, to membrane microdomains, called lipid rafts. A subpopulation of endogenous RGS16 in rat liver membranes and overexpressed RGS16 in COS cells, but not the nonpalmitoylated cysteine mutant of RGS16, localized to lipid rafts. However, disruption of lipid rafts by treatment with methyl-beta-cyclodextrin did not decrease the GTPase activating protein activity of RGS16. The lipid raft fractions were enriched in protein acyltransferase activity, and RGS16 incorporated [3H]palmitate into a peptide fragment containing Cys-98, a highly conserved cysteine within the RGS box. These results suggest that the amino-terminal palmitoylation of an RGS protein promotes its lipid raft targeting that allows palmitoylation of a poorly accessible cysteine residue that we show in the accompanying article (Osterhout, J. L., Waheed, A. A., Hiol, A., Ward, R. J., Davey, P. C., Nini, L., Wang, J., Milligan, G., Jones, T. L. Z., and Druey, K. M. (2003) J. Biol. Chem. 278, 19309-19316) was critical for RGS16 and RGS4 GAP activity. PMID- 12642594 TI - Distinct rates of palmitate turnover on membrane-bound cellular and oncogenic H ras. AB - H-Ras displays dynamic cycles of GTP binding and palmitate turnover. GTP binding is clearly coupled to activation, but whether the palmitoylated COOH terminus participates in signaling, especially when constrained by membrane tethering, is unknown. As a way to compare COOH termini of membrane-bound, lipid-modified H Ras, palmitate removal rates were measured for various forms of H-Ras in NIH 3T3 cells. Depalmitoylation occurred slowly (t(1/2) approximately 2.4 h) in cellular (H-RasWT) or dominant negative (H-Ras17N) forms and more rapidly (t(1/2) approximately 1 h) in oncogenic H-Ras61L or H-RasR12,T59. Combining this data with GTP binding measurements, the palmitate half-life of H-Ras in the fully GTP bound state was estimated to be less than 10 min. Slow palmitate removal from cellular H-Ras was not explained by sequestration in caveolae, as neither cellular nor oncogenic H-Ras showed alignment with caveolin by immunofluorescence. Conversely, although it had faster palmitate removal, oncogenic H-Ras was located in the same fractions as H-RasWT on four types of density gradients, and remained fully membrane-bound. Thus the different rates of deacylation occurred even though oncogenic and cellular H-Ras appeared to be in similar locations. Instead, these results suggest that acylprotein thioesterases access oncogenic H-Ras more easily because the conformation of its COOH terminus against the membrane is altered. This previously undetected difference could help produce distinctive effector interactions and signaling of oncogenic H-Ras. PMID- 12642595 TI - Growth hormone-induced phosphorylation of epidermal growth factor (EGF) receptor in 3T3-F442A cells. Modulation of EGF-induced trafficking and signaling. AB - Growth hormone (GH) promotes signaling by causing activation of the non-receptor tyrosine kinase, JAK2, which associates with the GH receptor. GH causes phosphorylation of epidermal growth factor receptor (EGFR; ErbB-1) and its family member, ErbB-2. For EGFR, JAK2-mediated GH-induced tyrosine phosphorylation may allow EGFR to serve as a scaffold for GH signaling. For ErbB-2, GH induces serine/threonine phosphorylation that dampens basal and EGF-induced ErbB-2 kinase activation. We now further explore GH-induced EGFR phosphorylation in 3T3-F442A, a preadipocytic fibroblast cell line that expresses endogenous GH receptor, EGFR, and ErbB-2. Using a monoclonal antibody that recognizes ERK consensus site phosphorylation (PTP101), we found that GH caused PTP101-reactive phosphorylation of EGFR. This GH-induced EGFR phosphorylation was prevented by MEK1 inhibitors but not by a protein kinase C inhibitor. Although GH did not discernibly affect EGF-induced EGFR tyrosine phosphorylation, we observed by immunoblotting a substantial decrease of EGF-induced EGFR degradation in the presence of GH. Fluorescence microscopy studies indicated that EGF-induced intracellular redistribution of an EGFR-cyan fluorescent protein chimera was markedly reduced by GH cotreatment, in support of the immunoblotting results. Notably, protection from EGF-induced degradation and inhibition of EGF-induced intracellular redistribution afforded by GH were both prevented by a MEK1 inhibitor, suggesting a role for GH-induced ERK activation in regulating the trafficking itinerary of the EGF-stimulated EGFR. Finally, we observed augmentation of early aspects of EGF signaling (EGF-induced ERK2 activation and EGF-induced Cbl tyrosine phosphorylation) by GH cotreatment; the GH effect on EGF-induced Cbl tyrosine phosphorylation was also prevented by MEK1 inhibition. These data indicate that GH, by activating ERKs, can modulate EGF-induced EGFR trafficking and signaling and expand our understanding of mechanisms of cross-talk between the GH and EGF signaling systems. PMID- 12642596 TI - Mechanism of rectification in inward-rectifier K+ channels. AB - Rectification in inward-rectifier K+ channels is caused by the binding of intracellular cations to their inner pore. The extreme sharpness of this rectification reflects strong voltage dependence (apparent valence is approximately 5) of channel block by long polyamines. To understand the mechanism by which polyamines cause rectification, we examined IRK1 (Kir2.1) block by a series of bis-alkyl-amines (bis-amines) and mono-alkyl-amines (mono-amines) of varying length. The apparent affinity of channel block by both types of alkylamines increases with chain length. Mutation D172N in the second transmembrane segment reduces the channel's affinity significantly for long bis amines, but only slightly for short ones (or for mono-amines of any length), whereas a double COOH-terminal mutation (E224G and E299S) moderately reduces the affinity for all bis-amines. The apparent valence of channel block increases from approximately 2 for short amines to saturate at approximately 5 for long bis amines or at approximately 4 for long mono-amines. On the basis of these and other observations, we propose that to block the channel pore one amine group in all alkylamines tested binds near the same internal locus formed by the COOH terminus, while the other amine group of bis-amines, or the alkyl tail of mono amines, "crawls" toward residue D172 and "pushes" up to 4 or 5 K+ ions outwardly across the narrow K+ selectivity filter. The strong voltage dependence of channel block therefore reflects the movement of charges carried across the transmembrane electrical field primarily by K+ ions, not by the amine molecule itself, as K+ ions and the amine blocker displace each other during block and unblock of the pore. This simple displacement model readily accounts for the classical observation that, at a given concentration of intracellular K+, rectification is apparently related to the difference between the membrane potential and the equilibrium potential for K+ ions rather than to the membrane potential itself. PMID- 12642597 TI - Simulation of calcium sparks in cut skeletal muscle fibers of the frog. AB - Spark mass, the volume integral of Delta F/F, was investigated theoretically and with simulations. These studies show that the amount of Ca2+ bound to fluo-3 is proportional to mass times the total concentration of fluo-3 ([fluo-3T]); the proportionality constant depends on resting Ca2+ concentration ([Ca2+]R). In the simulation of a Ca2+ spark in an intact frog fiber with [fluo-3T] = 100 microM, fluo-3 captures approximately one-fourth of the Ca2+ released from the sarcoplasmic reticulum (SR). Since mass in cut fibers is several times that in intact fibers, both with similar values of [fluo-3T] and [Ca2+]R, it seems likely that SR Ca2+ release is larger in cut fiber sparks or that fluo-3 is able to capture a larger fraction of the released Ca2+ in cut fibers, perhaps because of reduced intrinsic Ca2+ buffering. Computer simulations were used to identify these and other factors that may underlie the differences in mass and other properties of sparks in intact and cut fibers. Our spark model, which successfully simulates calcium sparks in intact fibers, was modified to reflect the conditions of cut fiber measurements. The results show that, if the protein Ca2+-buffering power of myoplasm is the same as that in intact fibers, the Ca2+ source flux underlying a spark in cut fibers is 5-10 times that in intact fibers. Smaller source fluxes are required for less buffer. In the extreme case in which Ca2+ binding to troponin is zero, the source flux needs to be 3-5 times that in intact fibers. An increased Ca2+ source flux could arise from an increase in Ca2+ flux through one ryanodine receptor (RYR) or an increase in the number of active RYRs per spark, or both. These results indicate that the gating of RYRs, or their apparent single channel Ca2+ flux, is different in frog cut fibers--and, perhaps, in other disrupted preparations--than in intact fibers. PMID- 12642598 TI - The pore region of the skeletal muscle ryanodine receptor is a primary locus for excitation-contraction uncoupling in central core disease. AB - Human central core disease (CCD) is caused by mutations/deletions in the gene that encodes the skeletal muscle ryanodine receptor (RyR1). Previous studies have shown that CCD mutations in the NH2-terminal region of RyR1 lead to the formation of leaky SR Ca2+ release channels when expressed in myotubes derived from RyR1 knockout (dyspedic) mice, whereas a COOH-terminal mutant (I4897T) results in channels that are not leaky to Ca2+ but lack depolarization-induced Ca2+ release (termed excitation-contraction [EC] uncoupling). We show here that store depletion resulting from NH2-terminal (Y523S) and COOH-terminal (Y4795C) leaky CCD mutant release channels is eliminated after incorporation of the I4897T mutation into the channel (Y523S/I4897T and Y4795C/I4897T). In spite of normal SR Ca2+ content, myotubes expressing the double mutants lacked voltage-gated Ca2+ release and thus exhibited an EC uncoupling phenotype similar to that of I4897T expressing myotubes. We also show that dyspedic myotubes expressing each of seven recently identified CCD mutations located in exon 102 of the RyR1 gene (G4890R, R4892W, I4897T, G4898E, G4898R, A4905V, R4913G) behave as EC-uncoupled release channels. Interestingly, voltage-gated Ca2+ release was nearly abolished (reduced approximately 90%) while caffeine-induced Ca2+ release was only marginally reduced in R4892W-expressing myotubes, indicating that this mutation preferentially disrupts voltage-sensor activation of release. These data demonstrate that CCD mutations in exon 102 disrupt release channel permeation to Ca2+ during EC coupling and that this region represents a primary molecular locus for EC uncoupling in CCD. PMID- 12642599 TI - Decoy receptors in the regulation of T helper cell type 2 responses. PMID- 12642600 TI - Directing autoimmunity to nucleoprotein particles: the impact of dendritic cells and interferon alpha in lupus. PMID- 12642601 TI - Regulation and function of the interleukin 13 receptor alpha 2 during a T helper cell type 2-dominant immune response. AB - Highly polarized type 2 cytokine responses can be harmful and even lethal to the host if they are too vigorous or persist too long. Therefore, it is important to elucidate the mechanisms that down-regulate these reactions. Interleukin (IL)-13 has emerged as a central mediator of T helper cell (Th)2-dominant immune responses, exhibiting a diverse array of functional activities including regulation of airway hyperreactivity, resistance to nematode parasites, and tissue remodeling and fibrosis. Here, we show that IL-13 receptor (R)alpha2 is a critical down-regulatory factor of IL-13-mediated tissue fibrosis induced by the parasitic helminth Schistosoma mansoni. IL-13Ralpha2 expression was induced after the onset of the fibrotic response, IL-10, IL-13, and Stat6 dependent, and inhibited by the Th1-inducing adjuvant IL-12. Strikingly, schistosome-infected C57BL/6 and BALB/c IL-13Ralpha2-deficient mice showed a marked exacerbation in hepatic fibrosis, despite displaying no change in granuloma size, tissue eosinophilia, or mastocytosis. Fibrosis increased despite the fact that IL-13 levels decreased significantly in the liver and serum. Importantly, pathology was prevented when IL-13Ralpha2-deficient mice were treated with a soluble IL 13Ralpha2-Fc construct, formally demonstrating that their exacerbated fibrotic response was due to heightened IL-13 activity. Together, these studies illustrate the central role played by the IL-13Ralpha2 in the down-regulation of a chronic and pathogenic Th2-mediated immune response. PMID- 12642603 TI - Interferon and granulopoiesis signatures in systemic lupus erythematosus blood. AB - Systemic lupus erythematosus (SLE) is a prototype systemic autoimmune disease characterized by flares of high morbidity. Using oligonucleotide microarrays, we now show that active SLE can be distinguished by a remarkably homogeneous gene expression pattern with overexpression of granulopoiesis-related and interferon (IFN)-induced genes. Using the most stringent statistical analysis (Bonferroni correction), 15 genes were found highly up-regulated in SLE patients, 14 of which are targets of IFN and one, defensin DEFA-3, a major product of immature granulocytes. A more liberal correction (Benjamini and Hochberg correction) yielded 18 additional genes, 12 of which are IFN-regulated and 4 granulocyte specific. Indeed immature neutrophils were identified in a large fraction of SLE patients white blood cells. High dose glucocorticoids, a standard treatment of disease flares, shuts down the interferon signature, further supporting the role of this cytokine in SLE. The expression of 10 genes correlated with disease activity according to the SLEDAI. The most striking correlation (P < 0.001, r = 0.55) was found with the formyl peptide receptor-like 1 protein that mediates chemotactic activities of defensins. Therefore, while the IFN signature confirms the central role of this cytokine in SLE, microarray analysis of blood cells reveals that immature granulocytes may be involved in SLE pathogenesis. PMID- 12642602 TI - Enhanced interleukin (IL)-13 responses in mice lacking IL-13 receptor alpha 2. AB - Interleukin (IL)-13 has recently been shown to play important and unique roles in asthma, parasite immunity, and tumor recurrence. At least two distinct receptor components, IL-4 receptor (R)alpha and IL-13Ralpha1, mediate the diverse actions of IL-13. We have recently described an additional high affinity receptor for IL 13, IL-13Ralpha2, whose function in IL-13 signaling is unknown. To better appreciate the functional importance of IL-13Ralpha2, mice deficient in IL 13Ralpha2 were generated by gene targeting. Serum immunoglobulin E levels were increased in IL-13Ralpha2-/- mice despite the fact that serum IL-13 was absent and immune interferon gamma production increased compared with wild-type mice. IL 13Ralpha2-deficient mice display increased bone marrow macrophage progenitor frequency and decreased tissue macrophage nitric oxide and IL-12 production in response to lipopolysaccharide. These results are consistent with a phenotype of enhanced IL-13 responsiveness and demonstrate a role for endogenous IL-13 and IL 13Ralpha2 in regulating immune responses in wild-type mice. PMID- 12642604 TI - Detection of a functional hybrid receptor gammac/GM-CSFRbeta in human hematopoietic CD34+ cells. AB - A functional hybrid receptor associating the common gamma chain (gammac) with the granulocyte/macrophage colony-stimulating factor receptor beta (GM-CSFRbeta) chain is found in mobilized human peripheral blood (MPB) CD34+ hematopoietic progenitors, SCF/Flt3-L primed cord blood (CB) precursors (CBPr CD34+/CD56-), and CD34+ myeloid cell lines, but not in normal natural killer (NK) cells, the cytolytic NK-L cell line or nonhematopoietic cells. We demonstrated, using CD34+ TF1beta cells, which express an interleukin (IL)-15Ralpha/beta/gammac receptor, that within the hybrid receptor, the GM-CSFRbeta chain inhibits the IL-15 triggered gammac/JAK3-specific signaling controlling TF1beta cell proliferation. However, the gammac chain is part of a functional GM-CSFR, activating GM-CSF dependent STAT5 nuclear translocation and the proliferation of TF1beta cells. The hybrid receptor is functional in normal hematopoietic progenitors in which both subunits control STAT5 activation. Finally, the parental TF1 cell line, which lacks the IL-15Rbeta chain, nevertheless expresses both a functional hybrid receptor that controls JAK3 phosphorylation and a novel IL-15alpha/gammac/TRAF2 complex that triggers nuclear factor kappaB activation. The lineage-dependent distribution and function of these receptors suggest that they are involved in hematopoiesis because they modify transduction pathways that play a major role in the differentiation of hematopoietic progenitors. PMID- 12642606 TI - Vaccination against Neisseria meningitidis using three variants of the lipoprotein GNA1870. AB - Sepsis and meningitis caused by serogroup B meningococcus are devastating diseases of infants and young adults, which cannot yet be prevented by vaccination. By genome mining, we discovered GNA1870, a new surface-exposed lipoprotein of Neisseria meningitidis that induces high levels of bactericidal antibodies. The antigen is expressed by all strains of N. meningitidis tested. Sequencing of the gene in 71 strains representative of the genetic and geographic diversity of the N. meningitidis population, showed that the protein can be divided into three variants. Conservation within each variant ranges between 91.6 to 100%, while between the variants the conservation can be as low as 62.8%. The level of expression varies between strains, which can be classified as high, intermediate, and low expressors. Antibodies against a recombinant form of the protein elicit complement-mediated killing of the strains that carry the same variant and induce passive protection in the infant rat model. Bactericidal titers are highest against those strains expressing high yields of the protein; however, even the very low expressors are efficiently killed. The novel antigen is a top candidate for the development of a new vaccine against meningococcus. PMID- 12642607 TI - Further proof of the plasticity of adult stem cells and their role in tissue repair. AB - In this issue, De Bari et al. (2003) present elegant data to counter the recent claims that adult stem cells have a limited plasticity. Further, they provide evidence that adult stem cells can seek out damaged tissues and repair them. PMID- 12642605 TI - Type-I interferon receptor deficiency reduces lupus-like disease in NZB mice. AB - Indirect evidence suggests that type-I interferons (IFN-alpha/beta) play a significant role in the pathogenesis of lupus. To directly examine the contribution of these pleiotropic molecules, we created congenic NZB mice lacking the alpha-chain of IFN-alpha/betaR, the common receptor for the multiple IFN alpha/beta species. Compared with littermate controls, homozygous IFN-alpha/betaR deleted NZB mice had significantly reduced anti-erythrocyte autoantibodies, erythroblastosis, hemolytic anemia, anti-DNA autoantibodies, kidney disease, and mortality. These reductions were intermediate in the heterozygous-deleted mice. The disease-ameliorating effects were accompanied by reductions in splenomegaly and in several immune cell subsets, including B-1 cells, the major producers of anti-erythrocyte autoantibodies. Decreases of B and T cell proliferation in vitro and in vivo, and of dendritic cell maturation and T cell stimulatory activity in vitro were also detected. Absence of signaling through the IFN-alpha/betaR, however, did not affect increased basal levels of the IFN-responsive p202 phosphoprotein, encoded by a polymorphic variant of the Ifi202 gene associated with the Nba2 predisposing locus in NZB mice. The data indicate that type-I IFNs are important mediators in the pathogenesis of murine lupus, and that reducing their activity in the human counterpart may be beneficial. PMID- 12642608 TI - Polarized growth and organelle segregation in yeast: the tracks, motors, and receptors. AB - In yeast, growth and organelle segregation requires formin-dependent assembly of polarized actin cables. These tracks are used by myosin Vs to deliver secretory vesicles for cell growth, organelles for their segregation, and mRNA for fate determination. Several specific receptors have been identified that interact with the cargo-binding tails of the myosin Vs. A recent study implicates specific degradation in the bud of the vacuolar receptor, Vac17, as a mechanism for cell cycle-regulated segregation of this organelle. PMID- 12642609 TI - Axonal transport of membranous and nonmembranous cargoes: a unified perspective. AB - Membranous and nonmembranous cargoes are transported along axons in the fast and slow components of axonal transport, respectively. Recent observations on the movement of cytoskeletal polymers in axons suggest that slow axonal transport is generated by fast motors and that the slow rate is due to rapid movements interrupted by prolonged pauses. This supports a unified perspective for fast and slow axonal transport based on rapid movements of diverse cargo structures that differ in the proportion of the time that they spend moving. A Flash feature (http://www.jcb.org/cgi/content/full/jcb.200212017/DC1) accompanies this Mini Review. PMID- 12642610 TI - The RasGAP-associated endoribonuclease G3BP assembles stress granules. AB - Stress granules (SGs) are formed in the cytoplasm in response to various toxic agents, and are believed to play a critical role in the regulation of mRNA metabolism during stress. In SGs, mRNAs are stored in an abortive translation initiation complex that can be routed to either translation initiation or degradation. Here, we show that G3BP, a phosphorylation-dependent endoribonuclease that interacts with RasGAP, is recruited to SGs in cells exposed to arsenite. G3BP may thus determine the fate of mRNAs during cellular stress. Remarkably, SG assembly can be either dominantly induced by G3BP overexpression, or on the contrary, inhibited by expressing a central domain of G3BP. This region binds RasGAP and contains serine 149, whose dephosphorylation is induced by arsenite treatment. Critically, a phosphomimetic mutant (S149E) fails to oligomerize and to assemble SGs, whereas a nonphosphorylatable G3BP mutant (S149A) does both. These results suggest that G3BP is an effector of SG assembly, and that Ras signaling contributes to this process by regulating G3BP dephosphorylation. PMID- 12642611 TI - Differential kinetochore protein requirements for establishment versus propagation of centromere activity in Saccharomyces cerevisiae. AB - Dicentric chromosomes undergo a breakage-fusion-bridge cycle as a consequence of having two centromeres on the same chromatid attach to opposite spindle poles in mitosis. Suppression of dicentric chromosome breakage reflects loss of kinetochore function at the kinetochore-microtubule or the kinetochore-DNA interface. Using a conditionally functional dicentric chromosome in vivo, we demonstrate that kinetochore mutants exhibit quantitative differences in their degree of chromosome breakage. Mutations in chl4/mcm17/ctf17 segregate dicentric chromosomes through successive cell divisions without breakage, indicating that only one of the two centromeres is functional. Centromere DNA introduced into the cell is unable to promote kinetochore assembly in the absence of CHL4. In contrast, established centromeres retain their segregation capacity for greater than 25 generations after depletion of Chl4p. The persistent mitotic stability of established centromeres reveals the presence of an epigenetic component in kinetochore segregation. Furthermore, this study identifies Chl4p in the initiation and specification of a heritable chromatin state. PMID- 12642612 TI - PAR-dependent and geometry-dependent mechanisms of spindle positioning. AB - During intrinsically asymmetric division, the spindle is oriented onto a polarized axis specified by a group of conserved PAR proteins. Extrinsic geometric asymmetry generated by cell shape also affects spindle orientation in some systems, but how intrinsic and extrinsic mechanisms coexist without interfering with each other is unknown. In some asymmetrically dividing cells of the wild-type Caenorhabditis elegans embryo, nuclear rotation directed toward the anterior cortex orients the forming spindle. We find that in such cells, a PAR dependent mechanism dominates and causes rotation onto the polarized axis, regardless of cell shape. However, when geometric asymmetry is removed, free nuclear rotation in the center of the cell is observed, indicating that the anterior-directed nature of rotation in unaltered embryos is an effect of cell shape. This free rotation is inconsistent with the prevailing model for nuclear rotation, the specialized cortical site model. In contrast, in par-3 mutant embryos, a geometry-dependent mechanism becomes active and causes directed nuclear rotation. These results lead to the model that in wild-type embryos both PAR-3 and PAR-2 are essential for nuclear rotation in asymmetrically dividing cells, but that PAR-3 inhibits geometry-dependent rotation in nonpolarized cells, thus preventing cell shape from interfering with spindle orientation. PMID- 12642613 TI - Budding yeast PAK kinases regulate mitotic exit by two different mechanisms. AB - We report the characterization of the dominant-negative CLA4t allele of the budding yeast CLA4 gene, encoding a member of the p21-activated kinase (PAK) family of protein kinases, which, together with its homologue STE20, plays an essential role in promoting budding and cytokinesis. Overproduction of the Cla4t protein likely inhibits both endogenous Cla4 and Ste20 and causes a delay in the onset of anaphase that correlates with inactivation of Cdc20/anaphase-promoting complex (APC)-dependent proteolysis of both the cyclinB Clb2 and securin. Although the precise mechanism of APC inhibition by Cla4t remains to be elucidated, our results suggest that Cla4 and Ste20 may regulate the first wave of cyclinB proteolysis mediated by Cdc20/APC, which has been shown to be crucial for activation of the mitotic exit network (MEN). We show that the Cdk1 inhibitory kinase Swe1 is required for the Cla4t-dependent delay in cell cycle progression, suggesting that it might be required to prevent full Cdc20/APC and MEN activation. In addition, inhibition of PAK kinases by Cla4t prevents mitotic exit also by a Swe1-independent mechanism impinging directly on the MEN activator Tem1. PMID- 12642614 TI - Identification of an organelle-specific myosin V receptor. AB - Class V myosins are widely distributed among diverse organisms and move cargo along actin filaments. Some myosin Vs move multiple types of cargo, where the timing of movement and the destinations of selected cargoes are unique. Here, we report the discovery of an organelle-specific myosin V receptor. Vac17p, a novel protein, is a component of the vacuole-specific receptor for Myo2p, a Saccharomyces cerevisiae myosin V. Vac17p interacts with the Myo2p cargo-binding domain, but not with vacuole inheritance-defective myo2 mutants that have single amino acid changes within this region. Moreover, a region of the Myo2p tail required specifically for secretory vesicle transport is neither required for vacuole inheritance nor for Vac17p-Myo2p interactions. Vac17p is localized on the vacuole membrane, and vacuole-associated Myo2p increases in proportion with an increase in Vac17p. Furthermore, Vac17p is not required for movement of other cargo moved by Myo2p. These findings demonstrate that Vac17p is a component of a vacuole-specific receptor for Myo2p. Organelle-specific receptors such as Vac17p provide a mechanism whereby a single type of myosin V can move diverse cargoes to distinct destinations at different times. PMID- 12642615 TI - Drosophila paramyosin is important for myoblast fusion and essential for myofibril formation. AB - Paramyosin is a major structural protein of thick filaments in invertebrate muscles. Coiled-coil dimers of paramyosin form a paracrystalline core of these filaments, and the motor protein myosin is arranged on the core surface. To investigate the function of paramyosin in myofibril assembly and muscle contraction, we functionally disrupted the Drosophila melanogaster paramyosin gene by mobilizing a P element located in its promoter region. Homozygous paramyosin mutants die at the late embryo stage. Mutants display defects in both myoblast fusion and in myofibril assembly in embryonic body wall muscles. Mutant embryos have an abnormal body wall muscle fiber pattern arising from defects in myoblast fusion. In addition, sarcomeric units do not assemble properly and muscle contractility is impaired. We confirmed that these defects are paramyosin specific by rescuing the homozygous paramyosin mutant to adulthood with a paramyosin transgene. Antibody analysis of normal embryos demonstrated that paramyosin accumulates as a cytoplasmic protein in early embryo development before assembling into thick filaments. We conclude that paramyosin plays an unexpected role in myoblast fusion and is important for myofibril assembly and muscle contraction. PMID- 12642616 TI - A neurotoxic peripherin splice variant in a mouse model of ALS. AB - Peripherin, a neuronal intermediate filament (nIF) protein found associated with pathological aggregates in motor neurons of patients with amyotrophic lateral sclerosis (ALS) and of transgenic mice overexpressing mutant superoxide dismutase 1 (SOD1G37R), induces the selective degeneration of motor neurons when overexpressed in transgenic mice. Mouse peripherin is unique compared with other nIF proteins in that three peripherin isoforms are generated by alternative splicing. Here, the properties of the peripherin splice variants Per 58, Per 56, and Per 61 have been investigated in transfected cell lines, in primary motor neurons, and in transgenic mice overexpressing peripherin or overexpressing SOD1G37R. Of the three isoforms, Per 61 proved to be distinctly neurotoxic, being assembly incompetent and inducing degeneration of motor neurons in culture. Using isoform-specific antibodies, Per 61 expression was detected in motor neurons of SOD1G37R transgenic mice but not of control or peripherin transgenic mice. The Per 61 antibody also selectively labeled motor neurons and axonal spheroids in two cases of familial ALS and immunoprecipitated a higher molecular mass peripherin species from disease tissue. This evidence suggests that expression of neurotoxic splice variants of peripherin may contribute to the neurodegenerative mechanism in ALS. PMID- 12642618 TI - Introduction to cytoskeletal dynamics and pathfinding of neuronal growth cones. PMID- 12642617 TI - Formation of filopodia-like bundles in vitro from a dendritic network. AB - We report the development and characterization of an in vitro system for the formation of filopodia-like bundles. Beads coated with actin-related protein 2/3 (Arp2/3)-activating proteins can induce two distinct types of actin organization in cytoplasmic extracts: (1) comet tails or clouds displaying a dendritic array of actin filaments and (2) stars with filament bundles radiating from the bead. Actin filaments in these bundles, like those in filopodia, are long, unbranched, aligned, uniformly polar, and grow at the barbed end. Like filopodia, star bundles are enriched in fascin and lack Arp2/3 complex and capping protein. Transition from dendritic to bundled organization was induced by depletion of capping protein, and add-back of this protein restored the dendritic mode. Depletion experiments demonstrated that star formation is dependent on Arp2/3 complex. This poses the paradox of how Arp2/3 complex can be involved in the formation of both branched (lamellipodia-like) and unbranched (filopodia-like) actin structures. Using purified proteins, we showed that a small number of components are sufficient for the assembly of filopodia-like bundles: Wiskott Aldrich syndrome protein (WASP)-coated beads, actin, Arp2/3 complex, and fascin. We propose a model for filopodial formation in which actin filaments of a preexisting dendritic network are elongated by inhibition of capping and subsequently cross-linked into bundles by fascin. PMID- 12642619 TI - Regulation of growth cone actin dynamics by ADF/cofilin. AB - Nervous system development is reliant on neuronal pathfinding, the process in which axons are guided to their target cells by specific extracellular cues. The ability of neurons to extend over long distances in response to environmental guidance signals is made possible by the growth cone, a highly motile structure found at the end of neuronal processes. Growth cones detect directional cues and respond with either attractive or repulsive movements. The motility of growth cones is dependent on rapid reorganization of the actin cytoskeleton, presumably mediated by actin-associated proteins under the control of incoming guidance signals. This article reviews how one such family of proteins, the ADF/cofilins, are emerging as key regulators of growth cone actin dynamics. These proteins are essential for rapid actin turnover in a variety of different cell types. ADF/cofilins are heavily co-localized with actin in growth cones and are necessary for neurite outgrowth. ADF/cofilin activities are regulated through reversible phosphorylation by LIM kinases and slingshot phosphatases. LIM kinases are downstream effectors of the Rho GTPases Rho, Rac, and Cdc42. Growing evidence suggests that extracellular guidance cues may locally alter actin dynamics by regulating the activity of LIM kinase and ADF/cofilin phosphatases via the Rho GTPases. In this way, ADF/cofilins and their upstream effectors may be pivotal to our understanding of how guidance information is translated into physical alterations of the growth cone actin cytoskeleton. PMID- 12642620 TI - Role of myosin II in axon outgrowth. AB - The initial stages of nerve outgrowth carried out by growth cones occur in three fundamental cyclic steps. Each of these steps appears to require myosin II activity to variable degrees. The steps include the following: (a) exploration, involving extensions and retractions that are driven and controlled by the interaction of actin retrograde flow and polymerization; (b) adhesion of new extensions to the substrate, which has been shown to be mediated by complex interactions between extracellular matrix proteins, cell adhesion proteins, and the actin cytoskeleton; and (c) traction force generated during forward advance of the growth cone, resulting in the production of tension on the neurite. PMID- 12642621 TI - Regulation of growth cone extension by SNARE proteins. AB - Recent studies have suggested that the soluble N-ethylmaleimide-sensitive factor attached protein (SNAP) receptor (SNARE)-mediated membrane fusion system is involved in vesicle fusion with the surface plasma membrane, which leads to neurite elongation. There have been several reports analyzing the effects of neurite outgrowth by inhibition of SNAREs. We studied this mechanism by overexpressing GFP-fusion SNAREs including VAMP-2, SNAP-25A, and syntaxin1A in PC12 cells to investigate the role of SNAREs in neurite outgrowth. When overexpressed in PC12 cells, VAMP-2 promoted neurite elongation, whereas SNAP-25A stimulated neurite sprouting. On the other hand, overexpression of syntaxin1A neither promoted nor inhibited neurite outgrowth. Thus, VAMP-2 and SNAP-25A play different roles in neurite elongation and sprouting. PMID- 12642622 TI - Growth cones integrate signaling from multiple guidance cues. AB - Nerve growth factor (NGF) and semaphorin3A (Sema3A) are guidance cues found in pathways and targets of developing dorsal root ganglia (DRG) neurons. DRG growth cone motility is regulated by cytoplasmic signaling triggered by these molecules. We investigated interactions of NGF and Sema3A in modulating growth cone behaviors of axons extended from E7 chick embryo DRGs. Axons extending in collagen matrices were repelled by Sema3A released from transfected HEK293 cells. However, if an NGF-coated bead was placed adjacent to Sema3A-producing cells, axons converged at the NGF bead. Growth cones of DRGs raised in 10(-9) M NGF were more resistant to Sema3A-induced collapse than when DRGs were raised in 10(-11) M NGF. After overnight culture in 10(-11) M NGF, 1-hr treatment with 10(-9) M NGF also increased growth cone resistance to Sema3A. Pharmacological studies indicated that the activities of ROCK and PKG participate in the cytoskeletal alterations that lead to Sema3A-induced growth cone collapse, whereas PKA activity is required for NGF-mediated reduction of Sema3A-induced growth cone collapse. These results support the idea that growth cone responses to a guidance cue can be modulated by interactions involving coincident signaling by other guidance cues. PMID- 12642623 TI - Gradients and growth cone guidance of grasshopper neurons. AB - The generation of a functional nervous system is dependent on precise pathfinding of axons during development. This pathfinding is directed by the distribution of local and long-range guidance cues, the latter of which are believed to be distributed in gradients. Gradients of guidance cues have been associated with growth cone function for over a hundred years. However, little is known about the mechanisms used by growth cones to respond to these gradients, in part owing to the lack of identifiable gradients in vivo. In the developing grasshopper limb, two gradients of the semaphorin Sema-2a are necessary for correct neuronal pathfinding in vivo. The gradients are found in regions where growth cones make critical steering decisions. Observations of different growth cone behaviors associated with these gradients have provided some insights into how growth cones respond to them. Growth cones appear to respond more faithfully to changes in concentration, rather than absolute levels, of Sema-2a expression, whereas the absolute levels may regulate growth cone size. PMID- 12642624 TI - Aint/Tacc3 is highly expressed in proliferating mouse tissues during development, spermatogenesis, and oogenesis. AB - Aint was originally identified on the basis of its interaction in vitro with the aryl hydrocarbon nuclear receptor translocator (Arnt). Arnt is a common heterodimerization partner in the basic helix-loop-helix (bHLH)-PER-ARNT-SIM (PAS) protein family and is involved in diverse biological functions. These include xenobiotic metabolism, hypoxic response, and circadian rhythm. In addition, Arnt has a crucial role during development. Aint is a member of a growing family of transforming acidic coiled-coil (TACC) proteins and is the murine homologue of human TACC3. Here we report the spatiotemporal expression of Tacc3 mRNA and protein in embryonic, postnatally developing, and adult mouse tissues using in situ hybridization and immunocytochemistry. Tacc3 mRNA was highly expressed in proliferating cells of several organs during murine development. However, the only adult tissues expressing high levels were testis and ovary. Immunocytochemistry revealed that Tacc3 is a nuclear protein. Our results suggest that Tacc3 has an important role in murine development, spermatogenesis, and oogenesis. PMID- 12642625 TI - Biochemical and immunohistochemical evidence for a non-muscle myosin at the neuromuscular junction in bovine skeletal muscle. AB - We identified 220-kD protein in bovine skeletal muscle homogenate by affinity chromatography on an agarose column and subsequent SDS-PAGE. Peptide mass fingerprinting (MALDI mass spectrometry) and internal sequence analysis revealed that this protein has homology with several members of the myosin superfamily, particularly with human cardiac beta-myosin heavy chain (beta-MHC). A rabbit polyclonal antibody against the 220-kD protein specifically stained a 220-kD band in Western blots of skeletal muscle homogenate. Immunohistochemical experiments on cryostat sections demonstrated that in skeletal muscle this protein is exclusively localized at the neuromuscular junctions, no immunoreactivity being present at the myofibril level. Because of its relative homology with cardiac beta-MHC, we also investigated the distribution of the 220-kD protein in bovine heart. In cardiac fibers, 220-kD protein-related immunoreactivity was restricted to the intercalated disks, whereas myofibrils were completely devoid of specific immunoreactivity. This distribution pattern was completely different from that of cardiac beta-MHC, which involved myofibrils. Because of the above biochemical and immunohistochemical features, the 220-kD protein we have identified is suggested to be a novel member of the non-muscle (non-sarcomeric) myosin family. PMID- 12642626 TI - Expression of galectin-1 and galectin-3 in human fetal thyroid gland. AB - High levels of expression of galectin-1 and galectin-3, the beta-galactoside binding proteins, have been recently described in malignant thyroid tumors but not in adenomas nor in normal thyroid tissue. However, there are no data about the expression of these galectins during fetal thyroid development. In this study we analyzed immunohistochemically the presence of galectin-1 and galectin-3 in human fetal thyroid glands (16-37 weeks of gestation). Weak to moderate cytoplasmic staining for galectin-1 was observed in follicular cells of all fetal thyroids. Galectin-3 could not be detected in thyroid follicular cells of any fetal thyroid investigated. Both galectins were detected in stromal tissue, but staining for galectin-1 was more intense. The absence of galectin-3 in thyroid cells during fetal development suggests that galectin-3 is expressed de novo during malignant transformation of thyroid epithelium, and that galectin-1 could be considered an oncofetal antigen. The results obtained indicated potential roles for galectin-1 and galectin-3 during the investigated period of human fetal thyroid gland development. Both galectins might participate in developmental processes regarding stromal fetal thyroid tissue organization, whereas galectin-1 might have a function in thyroid epithelium maturation. PMID- 12642627 TI - Expression of hornerin in stratified squamous epithelium in the mouse: a comparative analysis with profilaggrin. AB - We have recently identified a novel protein named hornerin, the structural features of which are most similar to those of profilaggrin, an essential protein for keratinization of epidermal tissues. In this study we examined the expression of hornerin compared with that of profilaggrin in various mouse tissues. Hornerin was expressed in the upper epidermis of newborn mouse skin, as was profilaggrin. In addition, both hornerin and profilaggrin were expressed in the tongue, esophagus, and forestomach. In all four tissues, immunostaining for hornerin and profilaggrin showed a granular pattern, and most of the signals for the two proteins were co-localized on keratohyalin granules. This was confirmed by double immunoelectron microscopy. Within keratohyalin granules, hornerin was detected more frequently in the periphery, whereas profilaggrin was equally distributed. A quantitative RT-PCR revealed that both genes were expressed at highest levels in the forestomach and at the next highest levels in skin. Profilaggrin mRNA was most abundant in the forestomach. In skin, the amount of hornerin mRNA was more than fourfold greater than the amount of profilaggrin mRNA. These results form the basis for a better understanding of possible overlapping and/or differential functions of hornerin and profilaggrin. PMID- 12642628 TI - Human kallikrein 13 expression in normal tissues: an immunohistochemical study. AB - The human tissue kallikrein 13 gene (KLK13), encoding for hK13 protein, was recently cloned and characterized. Here we describe the immunohistochemical (IHC) localization of hK13 in normal human tissues and compare it with the expression of two other kallikreins, hK6 and hK10. We performed the streptavidin-biotin IHC method on 204 paraffin blocks from archival, current, and autopsy material prepared from almost every normal human tissue, using a polyclonal and a monoclonal hK13 antibody. The staining was cytoplasmic and both antibodies yielded similar results. The hK13 protein was revealed in a variety of tissues, mainly in glandular epithelia. Other epithelia that expressed hK13 included the urothelium, the spermatic epithelium, and the epithelium of the choroid plexus. hK13 was intensely immunoexpressed by some endocrine organs, such as the adenohypophysis, the thyroid gland, the parathyroid glands, the adrenal medulla, the Leydig cells of the testis, and the cells of the endocrine pancreas. Immunoreactivity was also observed in the primordial follicles, the corpus luteum, and sparse luteinized cells in the stroma of the ovary, the trophoblastic cells of the placenta, the Hassall's corpuscles of the thymus, and chondrocytes. Nerves and ganglia of the peripheral nervous system, and both neurons and glial cells in the central nervous system, were positive. In short, hK13 was expressed by many glandular epithelia, some endocrine organs, and some specialized epithelia and cells. Comparison of these data with hK6 and hK10 expression suggests that the three kallikreins have a similar IHC pattern in normal human tissues. PMID- 12642630 TI - Expression of the LIM proteins paxillin and Hic-5 in human tissues. AB - The LIM domain is a protein-protein interaction motif critically involved in a variety of fundamental biological processes, including cytoskeletal organization, cell lineage specification, and organ development. In this study we examined the expression of the LIM proteins paxillin and Hic-5 in adult human tissues by immunohistochemistry and immunoblotting. Paxillin expression was widespread and observed both in non-muscle and muscle tissues. Of the latter, paxillin was mainly expressed in multinuclear striated muscle. In contrast, Hic-5 showed restricted expression and was expressed in muscle tissues, mainly in mononuclear smooth muscle. Taken together with previous findings, it appears likely that the counterbalance between paxillin and Hic-5 may be deeply involved in muscle differentiation. PMID- 12642629 TI - Expression of the matricellular protein SPARC in murine lens: SPARC is necessary for the structural integrity of the capsular basement membrane. AB - SPARC (Secreted Protein, Acidic and Rich in Cysteine) is a matricellular glycoprotein that modulates cell proliferation, adhesion, migration, and extracellular matrix (ECM) production. Although SPARC is generally abundant in embryonic tissues and is diminished in adults, we have found that the expression of SPARC in murine lens persists throughout embryogenesis and adulthood. Our previous studies showed that targeted ablation of the SPARC gene in mice results in cataract formation, a pathology attributed partially to an abnormal lens capsule. Here we provide evidence that SPARC is not a structural component of the lens capsule. In contrast, SPARC is abundant in lens epithelial cells, and newly differentiated fiber cells, with stable expression in wild-type mice up to 2 years of age. Pertubation of the lens capsule in animals lacking SPARC appears to be a consequence of the invasion of the lens cells situated beneath the capsule. Immunoreactivity for SPARC in the lens cells was uneven, with minimal reactivity in the epithelial cells immediately anterior to the equator. These epithelial cells appeared essentially noninvasive in SPARC-null mice, in comparison to the centrally located anterior epithelial cells, in which strong labeling by anti SPARC IgG was observed. The posterior lens fibers exhibited cytoplasmic extensions into the posterior lens capsule, which was severely damaged in SPARC null lenses. The expression of SPARC in wild-type lens cells, together with the abnormal lens capsule in SPARC-null mice, indicated that the structural integrity of the lens capsule is dependent on the matricellular protein SPARC. The effects of SPARC in the lens appear to involve regulation of lens epithelial and fiber cell morphology and functions rather than deposition as a structural component of the lens capsule. PMID- 12642631 TI - New techniques for whole-mount NADPH-diaphorase histochemistry demonstrated in insect ganglia. AB - Fixation-resistant NADPH-diaphorase (NADPHd) activity is used widely as a marker for nitric oxide synthase (NOS). In frozen sections, NADPHd histochemistry yields high anatomic definition. In whole-mounts, however, poor penetration of the reagents, background staining, and tissue opacity severely limit its application. Here we report a combination of new methods that significantly improves whole mount NADPHd staining. We demonstrate these methods in the thoracic ganglia of a large insect, the locust Schistocerca gregaria, in which NADPHd has been analyzed previously using both whole-mounts and serial section reconstructions. The penetration of the staining reagents was markedly improved after fixation in methanol/formalin compared to phosphate-buffered formaldehyde. Methanol/formalin also reduced nonspecific NADPHd and enhanced the selective staining. Penetration was further enhanced by incubation regimens that exploit the temperature- or pH dependence of NADPHd. In combination with methanol/formalin fixation, this permitted staining to develop evenly throughout these comparatively large invertebrate ganglia. These improvements were complemented by a new clearing technique that preserves the NADPHd staining, gives excellent transparency, and avoids distortion of specimen morphology. The new methods revealed the three dimensional architecture of NADPHd expression in locust ganglia in unprecedented detail and may similarly improve whole-mount detection of NADPHd in other invertebrate and vertebrate preparations. PMID- 12642632 TI - The nuclear localization of NFkappaB and p53 is positively correlated with HPV16 E7 level in laryngeal squamous cell carcinoma. AB - The interaction between the HPV (human papilloma virus) 16 E7 and other cell growth factors, such as p53 and NFkappaB in laryngeal cancer is not clearly understood. The aim of this study was to examine the expression of these three proteins in tumor and non-tumor laryngeal tissues from patients with laryngeal squamous cell carcinoma. These three proteins were dominantly expressed in the nucleus and their levels were higher in the tumor tissue than in the non-tumor tissue, although the comparison between the tumor and non-tumor tissues of p53 staining did not reach significance. The intensity of the nuclear stain of E7 and p53 was stronger than that of p65, a subunit of NFkappaB. Correlation analysis revealed that there was a positive relationship between the level of HPV16 E7 and the expression of p65. The correlation between E7 and p53 was also significant, although to a lesser degree. The finding of nuclear localization of p65 suggests that NFkappaB is constantly activated in the laryngeal cancer cells, whereas the sequestration of p53 in the nucleus may represent a mutated form of p53, which is probably inactivated by HPV16 oncoproteins. In conclusion, this study suggests that the nuclear localization of NFkappaB and p53 may play a role in the development of human laryngeal squamous cell carcinoma infected with HPV16. PMID- 12642633 TI - Intracellular targeting of calmodulin mRNAs in primary hippocampal cells. AB - We investigated the intracellular distribution of the mRNAs corresponding to the three non-allelic CaM genes in cultured hippocampal cells by in situ hybridization with digoxigenin-labeled gene-specific riboprobes. In neurons the perikaryon was heavily stained and strong dendritic mRNA targeting was detected for all three CaM genes. The color labeling exhibited a punctate distribution, suggesting that CaM mRNAs are transported in RNA granules. Immunocytochemistry for S100 demonstrated that glial cells express CaM mRNAs at a very low level. A minority of the cultured cells were negative for either labeling. PMID- 12642634 TI - Localization of the BiP molecular chaperone with respect to endoplasmic reticulum foci containing the cystic fibrosis transmembrane conductance regulator in yeast. AB - Almost all secreted proteins pass through the endoplasmic reticulum (ER), an organelle that is equipped to tolerate and/or degrade misfolded proteins. We report here that yeast expressing the cystic fibrosis transmembrane conductance regulator (CFTR) concentrate the protein at defined sites in the ER membrane that are not necessarily enriched for the ER molecular chaperone BiP. We propose that these sites are Russell bodies, an ER subcompartment in which misfolded proteins are stored and can be targeted for degradation. PMID- 12642635 TI - Fluorescence in situ hybridization (FISH) on human chromosomes using photoprobe biotin-labeled probes. AB - Fluorescence in situ hybridization (FISH) on human chromosomes in meta- and interphase is a well-established technique in clinical and tumor cytogenetics and for studies of evolution and interphase architecture. Many different protocols for labeling the DNA probes used for FISH have been published. Here we describe for the first time the successful use of Photoprobe biotin-labeled DNA probes in FISH experiments. Yeast artificial chromosome (YAC) and whole chromosome painting (wcp) probes were tested. PMID- 12642636 TI - Postmenopausal hormones--therapy for symptoms only. PMID- 12642637 TI - Effects of estrogen plus progestin on health-related quality of life. AB - BACKGROUND: The Women's Health Initiative (WHI) and other clinical trials indicate that significant health risks are associated with combination hormone use. Less is known about the effect of hormone therapy on health-related quality of life. METHODS: The WHI randomly assigned 16,608 postmenopausal women 50 to 79 years of age (mean, 63) with an intact uterus at base line to estrogen plus progestin (0.625 mg of conjugated equine estrogen plus 2.5 mg of medroxyprogesterone acetate, in 8506 women) or placebo (in 8102 women). Quality of-life measures were collected at base line and at one year in all women and at three years in a subgroup of 1511 women. RESULTS: Randomization to estrogen plus progestin resulted in no significant effects on general health, vitality, mental health, depressive symptoms, or sexual satisfaction. The use of estrogen plus progestin was associated with a statistically significant but small and not clinically meaningful benefit in terms of sleep disturbance, physical functioning, and bodily pain after one year (the mean benefit in terms of sleep disturbance was 0.4 point on a 20-point scale, in terms of physical functioning 0.8 point on a 100-point scale, and in terms of pain 1.9 points on a 100-point scale). At three years, there were no significant benefits in terms of any quality-of-life outcomes. Among women 50 to 54 years of age with moderate-to severe vasomotor symptoms at base line, estrogen and progestin improved vasomotor symptoms and resulted in a small benefit in terms of sleep disturbance but no benefit in terms of the other quality-of-life outcomes. CONCLUSIONS: In this trial in postmenopausal women, estrogen plus progestin did not have a clinically meaningful effect on health-related quality of life. PMID- 12642638 TI - The effect of differential reproductive success on population genetic structure: correlations of life history with matrilines in humpback whales of the gulf of maine. AB - To examine whether demographic and life-history traits are correlated with genetic structure, we contrasted mtDNA lineages of individual humpback whales (Megaptera novaeangliae) with sighting and reproductive histories of female humpback whales between 1979 and 1995. Maternal lineage haplotypes were obtained for 323 whales, either from direct sequencing of the mtDNA control region (n = 159) or inferred from known relationships along matrilines from the sequenced sample of individuals (n = 164). Sequence variation in the 550 bp of the control region defined a total of 19 maternal lineage haplotypes that formed two main clades. Fecundity increased significantly over the study period among females of several lineages among the two clades. Individual maternal lineages and other clades were characterized by significant variation in fecundity. The detected heterogeneity of reproductive success has the potential to substantially affect the frequency and distribution of maternal lineages found in this population over time. There were significant yearly effects on adult resighting rates and calf survivorship based on examination of sighting histories with varying capture recapture probability models. These results indicate that population structure can be influenced by interactions or associations between reproductive success, genetic structure, and environmental factors in a natural population of long lived mammals. PMID- 12642639 TI - Maternal effects and the evolution of behavioral and morphological characters: a literature review indicates the importance of extended maternal care. AB - Using literature data on reciprocal crosses, I estimated the influence of maternal effects on morphological and behavioral traits and compared these effects between mammals, birds, insects with homogametic females, and butterflies. In birds and in both groups of insects, no detectable difference between the reciprocal hybrids was observed on average, showing that in these groups, the contribution of maternal effects to the difference between the parental lines is at least rather small. In contrast to the other groups, mammals showed a significant and large influence of maternal effects on the examined characters. The large maternal effects in mammals are probably due to the extended period of parental care during gestation and lactation. It is concluded that maternal effects contributing to differences between parental lines are only widespread and important in mammals. It should be noted that these results do not show that maternal effects are absent in other animals. In the three examined groups, maternal effects may only evolve much more slowly than traits influenced by nuclear genes. PMID- 12642640 TI - A Bayesian model for assessing the frequency of multiple mating in nature. AB - Many breeding systems have multiple mating, in which males or females mate with multiple partners. With the advent of molecular markers, it is now possible to detect multiple mating in nature. However, no model yet exists to effectively assess the frequency of multiple mating (f(mm))--the proportion of broods with at least two males (or females) genetically contributing--from limited genetic data. We present a single-sex model based on Bayes' rule that incorporates the numbers of loci, alleles, offspring, and genetic parents. Two genetic criteria for calculating f(mm) are considered: the proportion of broods with three or more paternal (or maternal) alleles at any one locus and the total number of haplotypes observed in each brood. The former criterion provides the most precise estimates of f(mm). The model enables the calculation of confidence intervals and allows mutations (or typing errors) to be incorporated into the calculation. Failure to account for mutations can result in overestimates of f(mm). The model can also utilize other biological data, such as behavioral observations during mating, thereby increasing the accuracy of the calculation as compared to previous models. For example, when two sires contribute equally to multiply mated broods, only three loci with five equally common alleles are required to provide estimates of f(mm) with high precision. We demonstrate the model with an example addressing the frequency of multiple paternity in small versus large clutches of the endangered Kemp's Ridley sea turtle (Lepidochelys kempi) and show that females that lay large clutches are more likely to have multiply mated. PMID- 12642641 TI - Strong founder effect in Drosophila pseudoobscura colonizing New Zealand from North America. AB - The North American native species Drosophila pseudoobscura was first identified in New Zealand in the last few decades. Here, we have studied the genetic consequences of its spread across the Pacific Ocean. Using 10 microsatellites that are highly variable in North American populations, we found that the New Zealand population has substantially fewer alleles, a much lower average heterozygosity, and significantly different allele frequencies at these loci. We have discussed the relative sensitivity of these parameters for detecting the founding event. X-linked loci were more strongly differentiated between continents than autosomal loci, as reflected by larger changes in allele frequencies and greater reductions in numbers of alleles and average heterozygosity. The severity of the genetic diversity loss supports a scenario of a few D. pseudoobscura females being introduced to New Zealand from North America. PMID- 12642642 TI - The identification and characterization of two dominant r1 haplotype-specific inhibitors of aleurone color in Zea mays. AB - We report the identification and characterization of two novel dominant inhibitors of aleurone color in Zea mays that interact with specific haplotypes of the r1 locus. One inhibitor locus, inr1 (inhibitor of r1 aleurone color 1), maps to the long arm of chromosome 10, distal to the TB-10L19 breakpoint and tightly linked to dull1, and the second inhibitor locus, inr2 (inhibitor of r1 aleurone color 2), maps to the long arm of chromosome 9. Dominant inhibitory alleles of inr1 and inr2 act by suppressing aleurone color conditioned by certain r1 haplotypes. Two haplotypes, R1-ch:Stadler and R1-Randolph, exhibit nearly complete suppression of aleurone color in the presence of inhibitory alleles of inr1 or inr2. Two members of the R1-d class of haplotypes, R1-d:Catspaw and R1 d:Arapaho, show partial suppression. Other haplotypes tested were not visibly affected. The response of r1 haplotypes to inhibitory inr1 and inr2 alleles provides another means of analyzing the complex behavior of the seed color components of r1 haplotypes. Possible mechanisms of action of inr1 and inr2 are discussed. PMID- 12642643 TI - A partial african ancestry for the creole cattle populations of the Caribbean. AB - Seventy-eight cattle samples from three Creole Caribbean islands and one Brazilian breed were analyzed for sequence variation in the hypervariable segment of the mitochondrial DNA control region. Seventy-three samples displayed Bos taurus haplotypes, and five samples exhibited haplotypes that were of Bos indicus ancestry. Phylogenetic analysis revealed that all sampled B. taurus sequences fell into two distinct clusters with separate African and European origins. European sequences were encountered in each population; however, the distribution of African haplotypes was uneven, with the highest proportion of African influence found in the Guadeloupe Creole. The reduced levels of African haplotypic variation within the Caribbean and Brazilian are consistent with prior founder effects. Additionally, genetic variation at three microsatellite loci illustrated African influence uniquely in the Guadeloupe Creole. Collectively, the data suggest that this African influence is, at least in part, attributable to the historical importation of African cattle to the Americas. Furthermore, alleles of B. indicus ancestry were detected at appreciable frequencies in all Caribbean Creole populations and may reflect zebu introgressions from either West Africa or the Indian subcontinent. PMID- 12642644 TI - Disparity in population differentiation of sex-linked and autosomal variation in sibling species of the Jaera albifrons (Isopoda) complex. AB - The genetic variation at four enzyme loci is described for 22 populations of three Jaera species--J. albifrons, J. ischiosetosa, and J. praehirsuta--in the J. albifrons complex (Crustacea, Isopoda) in Denmark. The variation at three of the loci is similar, with the allele frequency spectra close to each other in all three species. An evolutionary tree based on the variation at these three loci revealed that the populations from the different species are completely intermixed in the tree. This was supported by hierarchical F-statistics where the between-species component was zero. At a fourth locus, Gpi (glucose phosphate isomerase), the species differ substantially. This locus is sex linked in J. ischiosetosa, but in the two other species, J. albifrons and J. praehirsuta, it is either found on autosomes or is sex linked with a high recombination rate between the locus and the centromere. An evolutionary tree for this locus partitions the populations into separate groups and a hierarchical F-statistic has a between-species component of about 50%. The results are attributed to introgression with a higher rate for autosomes than for sex chromosomes. PMID- 12642645 TI - Complete nucleotide sequence of the Coturnix chinensis (blue-breasted quail) mitochondrial genome and a phylogenetic analysis with related species. AB - Coturnix chinensis (blue-breasted quail) has been classically grouped in Galliformes Phasianidae Coturnix, based on morphologic features and biochemical evidence. Since the blue-breasted quail has the smallest body size among the species of Galliformes, in addition to a short generation time and an excellent reproductive performance, it is a possible model fowl for breeding and physiological studies of the Coturnix japonica (Japanese quail) and Gallus gallus domesticus (chicken), which are classified in the same family as blue-breasted quail. However, since its phylogenetic position in the family Phasianidae has not been determined conclusively, the sequence of the entire blue-breasted quail mitochondria (mt) genome was obtained to provide genetic information for phylogenetic analysis in the present study. The blue-breasted quail mtDNA was found to be a circular DNA of 16,687 base pairs (bp) with the same genomic structure as the mtDNAs of Japanese quail and chicken, though it is smaller than Japanese quail and chicken mtDNAs by 10 bp and 88 bp, respectively. The sequence identity of all mitochondrial genes, including those for 12S and 16S ribosomal RNAs, between blue-breasted quail and Japanese quail ranged from 84.5% to 93.5%; between blue-breasted quail and chicken, sequence identity ranged from 78.0% to 89.6%. In order to obtain information on the phylogenetic position of blue breasted quail in Galliformes Phasianidae, the 2,184 bp sequence comprising NADH dehydrogenase subunit 2 and cytochrome b genes available for eight species in Galliformes [Japanese quail, chicken, Gallus varius (green junglefowl), Bambusicola thoracica (Chinese bamboo partridge), Pavo cristatus (Indian peafowl), Perdix perdix (gray partridge), Phasianus colchicus (ring-neck pheasant), and Tympanchus phasianellus (sharp-tailed grouse)] together with that of Aythya americana (redhead) were examined using a maximum likelihood (ML) method. The ML analyses on the first/second codon positions, the third codon positions, and amino acid sequence consistently demonstrated that blue-breasted quail and Japanese quail are in the same phylogenetic cluster. PMID- 12642647 TI - Inheritance and allelism of resistance to soybean mosaic virus in Zao18 soybean from China. AB - Soybean mosaic disease caused by soybean mosaic virus (SMV) occurs wherever soybean [Glycine max (L.) Merr.] is grown and is considered one of the most important soybean diseases in many areas of the world. Use of soybean cultivars with resistance to SMV is a very effective way of controlling the disease. China has rich soybean germplasm, but there is very limited information on genetics of SMV resistance in Chinese soybean germplasm and reaction of the resistance genes to SMV strains G1-G7. There also is no report on allelic relationships of resistance genes in Chinese soybeans with other named genes at the three identified loci Rsv1, Rsv3, and Rsv4. The objectives of this study were to examine reactions of Chinese soybean cultivar Zao18 to SMV strains G1-G3 and G5 G7, to reveal the inheritance of SMV resistance in Zao18 and to determine the allelic relationship of resistance genes in Zao18 with previously reported resistance genes. Zao18 was crossed with the SMV-susceptible cultivar Lee 68 to study the inheritance of resistance. Zao18 was also crossed with the resistant lines PI96983, L29, and V94-5152, which possess Rsv1, Rsv3, and Rsv4, respectively, to examine the allelic relationship between the genes in Zao18 and genes at these three loci. Our research results indicated that Zao18 possesses two independent dominant genes for SMV resistance, one of which is allelic to the Rsv3 locus; the other is allelic with Rsv1. The presence of both genes (Rsv1 and Rsv3) in Zao18 confers resistance to SMV strains G1-G7. PMID- 12642646 TI - A new search for restorer cytoplasm: the restorer cytoplasm for the gene ms10 most probably does not exist in maize. AB - Since the suggestion concerning the hypothesis of the existence of restorer cytoplasms for some of the known and currently available male sterile (ms) genes in maize, a relatively limited amount of research effort has been made in order to test the hypothesis. Considering the importance of such a phenomenon, we designed a large, two-part experiment to test the idea more seriously. In the first part, 50 randomly chosen, medium-late open-pollinated (OP) varieties of maize from the Zemun Polje (ZP) collection were tested for the presence of the restorer cytoplasm in some currently known ms-genes in maize (ms1, ms2, ms3, ms4, ms5, ms6, ms7, ms8, ms9, ms11, ms12, ms13, ms17, ms22, ms23, and ms24). In the second part, the whole ZP collection of maize germplasm (more than 4,000 entries) was tested for the presence of the restorer cytoplasm for the gene ms10. After the first basic screening of OP varieties, more than 70 nonsegregating "candidates" were identified; however, after additional screening of the collection and the direct testing with respective homozygous ms-testers, all of them showed segregation, indicating that the restorer cytoplasm does not exist, especially the gene ms10. While performing this experiment, we discovered almost a hundred sources of male sterile cytoplasm, which were distinguished by their overwhelming frequency of male sterile plants in segregating test progenies. PMID- 12642648 TI - An efficient algorithm for the additive kinship matrix. PMID- 12642649 TI - SelAction: software to predict selection response and rate of inbreeding in livestock breeding programs. PMID- 12642650 TI - PARENTE: computer program for parentage analysis. PMID- 12642656 TI - Lymphocyte surface thiol levels. AB - Recent studies have implicated reduced thiols (cysteine -SH) in the function of individual cell surface proteins. Studies presented here demonstrate that the overall level of reduced thiols on cell surface molecules differs on individual subsets of peripheral blood mononuclear cells and that these levels can be manipulated in vitro by altering the level of intracellular glutathione (iGSH). To quantitate cell surface thiols, we have developed a Hi-D (11-color) fluorescence-activated cell sorter method in which we covalently couple a fluorescent molecule, Alexa-maleimide, to free (reduced) -SH groups on proteins or other molecules exposed on the cell surface (exofacial membrane). In addition, to reveal changes in cell surface thiol levels in response to various in vitro treatments, we used a pair of fluorescent Alexa dyes with distinct excitation and emission spectra to stain the cells before and after treatments. These in vitro studies demonstrate that decreasing iGSH, by specifically inhibiting its synthesis, decreases cell surface molecule thiols (csm-SH) and that preventing loss of iGSH also prevents loss of csm-SH. However, examination of peripheral blood mononuclear cell subsets tested immediately after isolation from healthy or HIV-infected subjects failed to reveal a similar relationship between internal iGSH and csm-SH. Although there is a relatively wide variation between individuals in both csm-SH and iGSH, there is no correlation between median iGSH and csm-SH compared for 22 healthy and 36 HIV-infected subjects. Collectively, our findings indicate that local environment plays a greater role in determining the redox status of cell surface molecules than the internal redox status of the cells. PMID- 12642657 TI - Motion generation by Drosophila mechanosensory neurons. AB - In Drosophila melanogaster, hearing is supported by mechanosensory neurons transducing sound-induced vibrations of the antenna. It is shown here that these neurons additionally generate motions that mechanically drive the antenna and tune it to relevant sounds. Motion generation in the Drosophila auditory system is betrayed by the auditory mechanics; the antenna of the fly nonlinearly alters its tuning as stimulus intensity declines and oscillates spontaneously in the absence of sound. The susceptibility of auditory motion generation to mechanosensory mutations shows that motion is generated by mechanosensory neurons. Motion generation depends on molecular components of the mechanosensory transduction machinery (NompA, NompC, Btv, and TilB), apparently involving mechanical activity of ciliated dendrites and microtubule-dependent motors. Hence, in analogy to vertebrate hair cells, the mechanosensory neurons of the fly serve dual, transducing, and actuating roles, documenting a striking functional parallel between the vertebrate cochlea and the ears of Drosophila. PMID- 12642658 TI - Mitochondrial pathology and apoptotic muscle degeneration in Drosophila parkin mutants. AB - Parkinson's disease (PD) is a common neurodegenerative disorder characterized by loss of dopaminergic neurons in the substantia nigra. Several lines of evidence strongly implicate mitochondrial dysfunction as a major causative factor in PD, although the molecular mechanisms responsible for mitochondrial dysfunction are poorly understood. Recently, loss-of-function mutations in the parkin gene, which encodes a ubiquitin-protein ligase, were found to underlie a familial form of PD known as autosomal recessive juvenile parkinsonism (AR-JP). To gain insight into the molecular mechanism responsible for selective cell death in AR-JP, we have created a Drosophila model of this disorder. Drosophila parkin null mutants exhibit reduced lifespan, locomotor defects, and male sterility. The locomotor defects derive from apoptotic cell death of muscle subsets, whereas the male sterile phenotype derives from a spermatid individualization defect at a late stage of spermatogenesis. Mitochondrial pathology is the earliest manifestation of muscle degeneration and a prominent characteristic of individualizing spermatids in parkin mutants. These results indicate that the tissue-specific phenotypes observed in Drosophila parkin mutants result from mitochondrial dysfunction and raise the possibility that similar mitochondrial impairment triggers the selective cell loss observed in AR-JP. PMID- 12642659 TI - Ring-like pore structures of SecA: implication for bacterial protein-conducting channels. AB - SecA, an essential component of the general protein secretion pathway of bacteria, is present in Escherichia coli as soluble and membrane-integral forms. Here we show by electron microscopy that SecA assumes two characteristic forms in the presence of phospholipid monolayers: dumbbell-shaped elongated structures and ring-like pore structures. The ring-like pore structures with diameters of 8 nm and holes of 2 nm are found only in the presence of anionic phospholipids. These ring-like pore structures with larger 3- to 6-nm holes (without staining) were also observed by atomic force microscopic examination. They do not form in solution or in the presence of uncharged phosphatidylcholine. These ring-like phospholipid-induced pore-structures may form the core of bacterial protein conducting channels through bacterial membranes. PMID- 12642660 TI - A regulatory protein that interferes with activator-stimulated transcription in bacteria. AB - Transcriptional activator proteins in bacteria often operate by interaction with the C-terminal domain of the alpha-subunit of RNA polymerase (RNAP). Here we report the discovery of an "anti-alpha" factor Spx in Bacillus subtilis that blocks transcriptional activation by binding to the alpha-C-terminal domain, thereby interfering with the capacity of RNAP to respond to certain activator proteins. Spx disrupts complex formation between the activator proteins ResD and ComA and promoter-bound RNAP, and it does so by direct interaction with the alpha subunit. ResD- and ComA-stimulated transcription requires the proteolytic elimination of Spx by the ATP-dependent protease ClpXP. Spx represents a class of transcriptional regulators that inhibit activator-stimulated transcription by interaction with alpha. PMID- 12642661 TI - Antibody-enhanced pneumococcal adherence requires IgA1 protease. AB - IgA, the major class of Ig in secretions, classically functions by interfering with microbial attachment to host tissues. Many mucosal pathogens, including Streptococcus pneumoniae, express an IgA1 protease that may circumvent the protective effects of this Ig subclass. Because these proteases are specific for human IgA1, we generated human mAbs to the major surface antigen of the pneumococcus, its capsular polysaccharide, and tested their effect in a colonization model of bacterial adherence to respiratory epithelial cells in culture. Rather than inhibiting adherence, type-specific IgA1 markedly enhanced bacterial attachment to host cells, but only when cleaved by IgA1 protease. Neither antibodies of protease-insensitive subclasses (IgA2 and IgG) nor those directed against heterologous capsules had such activity. The adherence-promoting properties of cleaved antibodies correlated with the cationic characteristics of their variable segments, suggesting that bound Fab fragments may neutralize the inhibitory effect of negatively charged capsules on adhesive interaction with host cells. Coating of pneumococci with anticapsular polysaccharide antibody unmasked the bacterial phosphorylcholine ligand, allowing for increased adherence mediated by binding to the platelet activating factor receptor on epithelial cells. In addition, our findings provide evidence for a novel function of bacterial IgA1 proteases. These enzymes may enable pathogens to subvert the antigen specificity of the humoral immune response to facilitate adhesive interactions and persistence on the mucosal surface. PMID- 12642662 TI - Dealing with iron: common structural principles in proteins that transport iron and heme. AB - Iron is essential to life, but poses severe problems because of its toxicity and the insolubility of hydrated ferric ions at neutral pH. In animals, a family of proteins called transferrins are responsible for the sequestration, transport, and distribution of free iron. Comparison of the structure and function of transferrins with a completely unrelated protein hemopexin, which carries out the same function for heme, identifies molecular features that contribute to a successful protein system for iron acquisition, transport, and release. These include a two-domain protein structure with flexible hinges that allow these domains to enclose the bound ligand and provide suitable chemistry for stable binding and an appropriate trigger for release. PMID- 12642663 TI - Interconversion of two oxidized forms of taurine/alpha-ketoglutarate dioxygenase, a non-heme iron hydroxylase: evidence for bicarbonate binding. AB - Taurinealpha-ketoglutarate (alphaKG) dioxygenase, or TauD, is a mononuclear non heme iron hydroxylase that couples the oxidative decarboxylation of alphaKG to the decomposition of taurine, forming sulfite and aminoacetaldehyde. Prior studies revealed that taurine-free TauD catalyzes an O(2)- and alphaKG-dependent self-hydroxylation reaction involving Tyr-73, yielding an Fe(III)-catecholate chromophore with a lambda(max) of 550 nm. Here, a chromophore (lambda(max) 720 nm) is described and shown to arise from O(2)-dependent self-hydroxylation of TauD in the absence of alphaKG, but requiring the product succinate. A similar chromophore rapidly develops with the alternative oxidant H(2)O(2). Resonance Raman spectra indicate that the approximately 700-nm chromophore also arises from an Fe(III)-catecholate species, and site-directed mutagenesis studies again demonstrate Tyr-73 involvement. The approximately 700-nm and 550-nm species are shown to interconvert by the addition or removal of bicarbonate, consistent with the alphaKG-derived CO(2) remaining tightly bound to the oxidized metal site as bicarbonate. The relevance of the metal-bound bicarbonate in TauD to reactions of other members of this enzyme family is discussed. PMID- 12642664 TI - Electron-nuclear double resonance spectroscopy (and electron spin-echo envelope modulation spectroscopy) in bioinorganic chemistry. AB - This perspective discusses the ways that advanced paramagnetic resonance techniques, namely electron-nuclear double resonance (ENDOR) and electron spin echo envelope modulation (ESEEM) spectroscopies, can help us understand how metal ions function in biological systems. PMID- 12642665 TI - Bifunctional antisense oligonucleotides provide a trans-acting splicing enhancer that stimulates SMN2 gene expression in patient fibroblasts. AB - The multiplicity of proteins compared with genes in mammals owes much to alternative splicing. Splicing signals are so subtle and complex that small perturbations may allow the production of new mRNA variants. However, the flexibility of splicing can also be a liability, and several genetic diseases result from single-base changes that cause exons to be skipped during splicing. Conventional oligonucleotide strategies can block reactions but cannot restore splicing. We describe here a method by which the use of a defective exon was restored. Spinal muscular atrophy (SMA) results from mutations of the Survival Motor Neuron (SMN) gene. Mutations of SMN1 cause SMA, whereas SMN2 acts as a modifying gene. The two genes undergo alternative splicing with SMN1, producing an abundance of full-length mRNA transcripts, whereas SMN2 predominantly produces exon 7-deleted transcripts. This discrepancy is because of a single nucleotide difference in SMN2 exon 7, which disrupts an exonic splicing enhancer containing an SF2ASF binding site. We have designed oligoribonucleotides that are complementary to exon 7 and contain exonic splicing enhancer motifs to provide trans-acting enhancers. These tailed oligoribonucleotides increased SMN2 exon 7 splicing in vitro and rescued the incorporation of SMN2 exon 7 in SMA patient fibroblasts. This treatment also resulted in the partial restoration of gems, intranuclear structures containing SMN protein that are severely reduced in patients with SMA. The use of tailed antisense oligonucleotides to recruit positively acting factors to stimulate a splicing reaction may have therapeutic applications for genetic disorders, such as SMA, in which splicing patterns are altered. PMID- 12642666 TI - Impaired adrenocorticotropic hormone response to bacterial endotoxin in mice deficient in prostaglandin E receptor EP1 and EP3 subtypes. AB - Sickness evokes various neural responses, one of which is activation of the hypothalamo-pituitary-adrenal (HPA) axis. This response can be induced experimentally by injection of bacterial lipopolysaccharide (LPS) or inflammatory cytokines such as IL-1. Although prostaglandins (PGs) long have been implicated in LPS-induced HPA axis activation, the mechanism downstream of PGs remained unsettled. By using mice lacking each of the four PGE receptors (EP1-EP4) and an EP1-selective antagonist, ONO-8713, we showed that both EP1 and EP3 are required for adrenocorticotropic hormone release in response to LPS. Analysis of c-Fos expression as a marker for neuronal activity indicated that both EP1 and EP3 contribute to activation of neurons in the paraventricular nucleus of the hypothalamus (PVN). This analysis also revealed that EP1, but not EP3, is involved in LPS-induced activation of the central nucleus of the amygdala. EP1 immunostaining in the PVN revealed its localization at synapses on corticotropin releasing hormone-containing neurons. These findings suggest that EP1- and EP3 mediated neuronal pathways converge at corticotropin-releasing hormone-containing neurons in the PVN to induce HPA axis activation upon sickness. PMID- 12642667 TI - Convergent evolution of perenniality in rice and sorghum. AB - Annual and perennial habit are two major strategies by which grasses adapt to seasonal environmental change, and these distinguish cultivated cereals from their wild relatives. Rhizomatousness, a key trait contributing to perenniality, was investigated by using an F(2) population from a cross between cultivated rice (Oryza sativa) and its wild relative, Oryza longistaminata. Molecular mapping based on a complete simple sequence-repeat map revealed two dominant complementary genes controlling rhizomatousness. Rhz3 was mapped to the interval between markers OSR16 [1.3 centimorgans (cM)] and OSR13 (8.1 cM) on rice chromosome 4 and Rhz2 located between RM119 (2.2 cM) and RM273 (7.4 cM) on chromosome 3. Comparative mapping indicated that each gene closely corresponds to major quantitative trait loci (QTLs) controlling rhizomatousness in Sorghum propinquum, a wild relative of cultivated sorghum. Correspondence of these genes in rice and sorghum, which diverged from a common ancestor approximately 50 million years ago, suggests that the two genes may be key regulators of rhizome development in many Poaceae. Many additional QTLs affecting abundance of rhizomes in O. longistaminata were identified, most of which also corresponded to the locations of S. propinquum QTLs. Convergent evolution of independent mutations at, in some cases, corresponding genes may have been responsible for the evolution of annual cereals from perennial wild grasses. DNA markers closely linked to Rhz2 and Rhz3 will facilitate cloning of the genes, which may contribute significantly to our understanding of grass evolution, advance opportunities to develop perennial cereals, and offer insights into environmentally benign weed-control strategies. PMID- 12642668 TI - The role of disulfide bonds in the assembly and function of MD-2. AB - MD-2 is a secreted glycoprotein that binds to the extracellular domain of Toll like receptor 4 (TLR4) and is required for the activation of TLR4 by lipopolysaccharide (LPS). The protein contains seven Cys residues and consists of a heterogeneous collection of disulfide-linked oligomers. To investigate the role of sulfhydryls in MD-2 structure and function, we created 17 single and multiple Cys substitution mutants. All of the MD-2 mutant proteins, including one totally lacking Cys residues, were secreted and stable. SDSPAGE analyses indicated that most Cys residues could participate in oligomer formation and that no single Cys residue was required for oligomerization. Of the single Cys substitutions, only C95S and C105S failed to confer LPS responsiveness on TLR4 when mutant and TLR4 were cotransfected into cells expressing an NF-kappaB reporter plasmid. Surprisingly, substitution of both C95 and C105 partially restored activity. Structural analyses revealed that C95 and C105 formed an intrachain disulfide bond, whereas C95 by itself produced an inactive dimer. In contrast to the cotransfection experiments, only WT MD-2 conferred responsiveness to LPS when secreted proteins were added directly to TLR4 reporter cells. Our data are consistent with a model in which most, possibly all sulfhydryls lie on the surface of a stable MD-2 core structure where they form both intra- and interchain disulfide bridges. These disulfide bonds produce a heterogeneous array of oligomers, including some species that can form an active complex with TLR4. PMID- 12642669 TI - A duodenally absorbable CXC chemokine receptor 4 antagonist, KRH-1636, exhibits a potent and selective anti-HIV-1 activity. AB - A low molecular weight nonpeptide compound, KRH-1636, efficiently blocked replication of various T cell line-tropic (X4) HIV type 1 (HIV-1) in MT-4 cells and peripheral blood mononuclear cells through the inhibition of viral entry and membrane fusion via the CXC chemokine receptor (CXCR)4 coreceptor but not via CC chemokine receptor 5. It also inhibited binding of the CXC chemokine, stromal cell-derived factor 1alpha, to CXCR4 specifically and subsequent signal transduction. KRH-1636 prevented monoclonal antibodies from binding to CXCR4 without down-modulation of the coreceptor. The inhibitory effect against X4 viral replication by KRH-1636 was clearly reproduced in the human peripheral blood lymphocytesevere combined immunodeficiency mouse system. Furthermore, this compound was absorbed into the blood after intraduodenal administration as judged by anti-HIV-1 activity and liquid chromatography MS in the plasma. Thus, KRH-1636 seems to be a promising agent for the treatment of HIV-1 infection. PMID- 12642670 TI - Speculative synthetic chemistry and the nitrogenase problem. AB - There exist a limited but growing number of biological metal centers whose properties lie conspicuously outside the realm of known inorganic chemistry. The synthetic analogue approach, broadly directed, offers a powerful exploratory tool that can define intrinsic chemical possibilities for these sites while simultaneously expanding the frontiers of fundamental inorganic chemistry. This speculative application of analogue study is exemplified here in the evolution of synthetic efforts inspired by the cluster chemistry of biological nitrogen fixation. PMID- 12642671 TI - The organometallic active site of [Fe]hydrogenase: models and entatic states. AB - The simple organometallic, (mu-S(2))Fe(2)(CO)(6), serves as a precursor to synthetic analogues of the chemically rudimentary iron-only hydrogenase enzyme active site. The fundamental properties of the (mu SCH(2)CH(2)CH(2)S)[Fe(CO)(3)](2) compound, including structural mobility and regioselectivity in cyanidecarbon monoxide substitution reactions, relate to the enzyme active site in the form of transition-state structures along reaction paths rather than ground-state structures. Even in the absence of protein-based active-site organization, the ground-state structural model complexes are shown to serve as hydrogenase enzyme reaction models, H(2) uptake and H(2) production, with the input of photo- or electrochemical energy, respectively. PMID- 12642672 TI - Electrochemical and NMR spectroscopic studies of distal pocket mutants of nitrophorin 2: stability, structure, and dynamics of axial ligand complexes. AB - WT and leucine --> valine distal pocket mutants of nitrophorin 2 (NP2) and their NO complexes have been investigated by spectroelectrochemistry. NO complexes of two of the mutants exhibit more positive reduction potential shifts than does the WT protein, thus indicating stabilization of the Fe(II)-NO state. This more positive reduction potential for NP2-L132V and the double mutant is consistent with the hypothesis that smaller valine residues may allow the heme to regain planarity instead of being significantly ruffled, as in WT NP2. Thus, ruffling may stabilize the Fe(III)-NO state, which is required for facile NO dissociation. NMR spectroscopic investigations show that the sterically demanding 2 methylimidazole ligand readily binds to all three distal pocket mutants to create low-spin Fe(III) complexes having axial ligands in nearly perpendicular planes; it also binds to the WT protein in the presence of higher concentrations of 2 methylimidazole, but yields a different ligand plane orientation than is present in any of the three distal pocket mutants. NOESY spectra of NP2-ImH mutants exhibit chemical exchange cross peaks, whereas WT NP2-ImH shows no chemical exchange. Chemical exchange in the case of the distal leucine --> valine mutants is caused by ImH ligand orientational dynamics. The two angular orientations of the ImH ligand could be determined from the (1)H chemical shifts of the heme methyls, and the rate of interconversion of the two forms could be estimated from the NOESY diagonal and cross peak intensities. K(eq) is 100 or larger and favors an orientation similar to that found for the WT NP2-ImH complex. PMID- 12642674 TI - Regulation of Dlx3 gene expression in visceral arches by evolutionarily conserved enhancer elements. AB - The mammalian Distal-less (Dlx) clusters (Dlx1-2, Dlx5-6, and Dlx3-7) have a nested expression pattern in developing visceral (branchial) arches. Genetic regulatory mechanisms controlling Dlx spatial expression within the visceral arches have not yet been defined. Here we show that an enhancer in the Dlx3-7 cluster can regulate the visceral arch specific expression pattern of the Dlx3 gene. We have used a 79-kb transgene construct containing the entire Dlx3-7 bigene cluster with a LacZ reporter inserted in frame in the first exon of the Dlx3 gene. Visceral arch expression is absent when a 4-kb element located within the Dlx3-7 intergenic region is deleted. A 245-bp element (I37-2) whose DNA sequence is highly conserved between human and mouse located within the 4kb deleted region can drive visceral arch expression when fused to a hsp68-lacZ reporter transgene construct. Reporter expression is detected in 9.5 and 10.5 days postcoitum transgenic embryos in a manner consistent with the endogenous Dlx3 expression pattern in the mesenchyme of the first and second visceral arches. Thus the I37-2 element is both necessary and sufficient for Dlx3 expression. The I37-2 element contains several putative binding sites for several transcription factors including Dlx and other homeodomain proteins within the evolutionarily conserved region. Significantly, the I37-2 element shows a sequence-match including a Dlx binding site to a cis-element in the Dlx5-6 intermediate region designated mI56i [Zerucha, T., Stuhmer, T., Hatch, G., Park, B. K., Long, Q., Yu, G., Gambarotta, A., Schultz, J. R., Rubenstein, J. L. & Ekker, M. (2000) J. Neurosci. 20, 709-721], despite distant phylogenetic relationship between these clusters. Our results provide evidence for a concerted role for DLX auto- and cross-regulation in the establishment of a nested expression pattern for Dlx3-7 and Dlx5-6 clusters within the visceral arches. PMID- 12642673 TI - ARC5, a cytosolic dynamin-like protein from plants, is part of the chloroplast division machinery. AB - Chloroplast division in plant cells is orchestrated by a complex macromolecular machine with components positioned on both the inner and outer envelope surfaces. The only plastid division proteins identified to date are of endosymbiotic origin and are localized inside the organelle. Employing positional cloning methods in Arabidopsis in conjunction with a novel strategy for pinpointing the mutant locus, we have identified a gene encoding a new chloroplast division protein, ARC5. Mutants of ARC5 exhibit defects in chloroplast constriction, have enlarged, dumbbell-shaped chloroplasts, and are rescued by a wild-type copy of ARC5. The ARC5 gene product shares similarity with the dynamin family of GTPases, which mediate endocytosis, mitochondrial division, and other organellar fission and fusion events in eukaryotes. Phylogenetic analysis showed that ARC5 is related to a group of dynamin-like proteins unique to plants. A GFP-ARC5 fusion protein localizes to a ring at the chloroplast division site. Chloroplast import and protease protection assays indicate that the ARC5 ring is positioned on the outer surface of the chloroplast. Thus, ARC5 is the first cytosolic component of the chloroplast division complex to be identified. ARC5 has no obvious counterparts in prokaryotes, suggesting that it evolved from a dynamin-related protein present in the eukaryotic ancestor of plants. These results indicate that the chloroplast division apparatus is of mixed evolutionary origin and that it shares structural and mechanistic similarities with both the cell division machinery of bacteria and the dynamin-mediated organellar fission machineries of eukaryotes. PMID- 12642675 TI - Real-time semantic compensation in patients with agrammatic comprehension: electrophysiological evidence for multiple-route plasticity. AB - To understand spoken language requires that the brain provides rapid access to different kinds of knowledge, including the sounds and meanings of words, and syntax. Syntax specifies constraints on combining words in a grammatically well formed manner. Agrammatic patients are deficient in their ability to use these constraints, due to a lesion in the perisylvian area of the language-dominant hemisphere. We report a study on real-time auditory sentence processing in agrammatic comprehenders, examining their ability to accommodate damage to the language system. We recorded event-related brain potentials (ERPs) in agrammatic comprehenders, nonagrammatic aphasics, and age-matched controls. When listening to sentences with grammatical violations, the agrammatic aphasics did not show the same syntax-related ERP effect as the two other subject groups. Instead, the waveforms of the agrammatic aphasics were dominated by a meaning-related ERP effect, presumably reflecting their attempts to achieve understanding by the use of semantic constraints. These data demonstrate that although agrammatic aphasics are impaired in their ability to exploit syntactic information in real time, they can reduce the consequences of a syntactic deficit by exploiting a semantic route. They thus provide evidence for the compensation of a syntactic deficit by a stronger reliance on another route in mapping sound onto meaning. This is a form of plasticity that we refer to as multiple-route plasticity. PMID- 12642676 TI - Cell and tumor classification using gene expression data: construction of forests. AB - The advent of gene chips has led to a promising technology for cell, tumor, and cancer classification. We exploit and expand the methodology of recursive partitioning trees for tumor and cell classification from microarray gene expression data. To improve classification and prediction accuracy, we introduce a deterministic procedure to form forests of classification trees and compare their performance with extant alternatives. When two published and commonly used data sets are used, we find that the deterministic forests perform similarly to the random forests in terms of the error rate obtained from the leave-one-out procedure, and all of the forests are far better than the single trees. In addition, we provide graphical presentations to facilitate interpretation of complex forests and compare our findings with the current biological literature. In addition to numerical improvement, the main advantage of deterministic forests is reproducibility and scientific interpretability of all steps in tree construction. PMID- 12642677 TI - Cloning and engineering of the cinnamycin biosynthetic gene cluster from Streptomyces cinnamoneus cinnamoneus DSM 40005. AB - Lantibiotics are ribosomally synthesized oligopeptide antibiotics that contain lanthionine bridges derived by the posttranslational modification of amino acid residues. Here, we describe the cinnamycin biosynthetic gene cluster (cin) from Streptomyces cinnamoneus cinnamoneus DSM 40005, the first, to our knowledge, lantibiotic gene cluster from a high G+C bacterium to be cloned and sequenced. The cin cluster contains many genes not found in lantibiotic clusters from low G+C Gram-positive bacteria, including a Streptomyces antibiotic regulatory protein regulatory gene, and lacks others found in such clusters, such as a LanT type transporter and a LanP-type protease. Transfer of the cin cluster to Streptomyces lividans resulted in heterologous production of cinnamycin. Furthermore, modification of the cinnamycin structural gene (cinA) led to production of two naturally occurring lantibiotics, duramycin and duramycin B, closely resembling cinnamycin, whereas attempts to make a more widely diverged derivative, duramycin C, failed to generate biologically active material. These results provide a basis for future attempts to construct extensive libraries of cinnamycin variants. PMID- 12642678 TI - Statistically relevant conserved quantities for truncated quasigeostrophic flow. AB - Systematic applications of ideas from equilibrium statistical mechanics lead to promising strategies for assessing the unresolved scales of motion in many problems in science and engineering. A scientific debate over more than the last 25 years involves which conserved quantities among the formally infinite list are statistically relevant for the large-scale equilibrium statistical behavior. Here this important issue is addressed by using suitable discrete numerical approximations for geophysical flows with many conserved quantities as a numerical laboratory. The results of numerical experiments are presented here for these truncated geophysical flows with topography in a suitable regime. These experiments establish that the integrated third power of potential vorticity besides the familiar constraints of energy, circulation, and enstrophy (the integrated second power) is statistically relevant in this regime for the coarse grained equilibrium statistical behavior at large scales. Furthermore, the integrated higher powers of potential vorticity larger than three are statistically irrelevant for the large-scale equilibrium statistical behavior in the examples studied here. PMID- 12642681 TI - An introduction to pancreatic adenocarcinoma genetics, pathology and therapy. AB - Cancer is bad, but pancreatic cancer is especially so. Underlying the aggressive behavior of pancreatic cancer is a highly patterned set of biologic rules for its development. The accumulating discoveries regarding the genetic basis of pancreatic cancer and developments in the clinical management of the disease provide hope for meaningful improvements in the understanding and control of this disease. PMID- 12642680 TI - Ras family signaling: therapeutic targeting. AB - Mutationally activated and oncogenic versions of the ras genes were first identified in human tumors in 1982. This discovery prompted great interest in the development of anti-Ras strategies as novel, target-based approaches for cancer treatment. The three human ras genes represent the most frequently mutated oncogenes in human cancers. Consequently, a considerable research effort has been made to define the function of Ras in normal and neoplastic cells and to target Ras for cancer treatment. Among the anti-Ras strategies that are under evaluation in the clinic are pharmacologic inhibitors designed to prevent: (1) association with the plasma membrane (farnesyltransferase inhibitors), (2) downstream signaling (Raf and MEK protein kinase inhibitors), (3) autocrine growth factor signaling (EGF receptor inhibitors), or (4) gene expression (H-ras and c-raf-1). Although a number of these inhibitors have demonstrated potent anti-tumor activities in preclinical models, phase l-lll clinical trials have revealed unexpected complexities in Ras function and in the clinical development of target based therapies. We review the current status of anti-Ras drug development, issues that have complicated their progression to the clinic, and possible future strategies for targeting Ras. PMID- 12642682 TI - Treatment of acute promyelocytic leukemia with ATRA and As2O3: a model of molecular target-based cancer therapy. AB - Most acute promyelocytic leukemia (APL) cases have t(15;17)(q22;q21) chromosomal translocation and PML-RARalpha chimeric gene which blocks granulocytic differentiation. The introduction of all-trans-retinoic acid (ATRA) and arsenic compounds, especially arsenic trioxide (As(2)O(3)), has provided good models to study not only differentiation and/or apoptosis therapy but also molecular target based cancer treatment. In vivo and in vitro investigations have shown that both agents are able to induce differentiation of APL cells: ATRA tends to induce terminal differentiation, while low-dose As(2)O(3) can induce partial differentiation. Significant progress has been made in understanding the molecular mechanisms of APL pathogenesis and differentiation therapy. Pharmacological concentrations (0.1 approximately 1 microM) of ATRA derepresses transcription by releasing CoR from, and recruiting CoA to PML-RARalpha, whereas As(2)O(3) triggers a rapid degradation of PML-RARalpha. In fact, the two drugs act on the same oncoprotein through targeting different moieties and in distinct ways and thereby abrogate its dominant-negative effects on regulatory pathways necessary for granulocytic differentiation. As to apoptosis, it is clear that high-dose As(2)O(3) can induce mitochondria-mediated cell death pathway in a thiol-dependent manner, while the mechanism of ATRA-induced apoptosis needs further elucidation. Transcriptomic and proteomic analysis are also expected to find new molecular targets. It is the hope that what we have learnt from APL will benefit further developments of anti-leukemia therapy. PMID- 12642683 TI - beta-Catenin signaling: therapeutic strategies in oncology. AB - Activated Wnt signaling pathways have been found in various human cancers, including those of the colon, liver, endometrium, ovary, prostate, and stomach. As a result, beta-catenin is accumulated and becomes transcriptionally active for proliferative genes and oncogenes. Wnt pathway mutations result in biochemical mechanisms yielding inefficient phosphorylation of beta-catenin by GSK3beta due to APC, beta-catenin and/or axin mutations. Therefore, the needs and the opportunity to develop new cancer therapies exist through reversing oncogenic APC/beta-catenin/Lef/Tcf signals. Exisulind and analogues are inhibitors of cyclic GMP phosphodiesterases (PDE) that have been shown to activate and induce protein kinase G. The data show PKG regulation of beta-catenin in wnt signaling, accounting, at least in part, for apoptosis induction in treated colon cancer cells carrying either APC or beta-catenin mutations. Exisulind and analogs reduce beta-catenin via a novel, GSK3beta independent processing mechanism. Activated PKG directly phosphorylate beta-catenin at its C-terminal domain and causes proteasome dependent degradation of the protein. Since this pathway is independent of APC and GSK3beta, exisulind and analogs provide a superior approach to circumvent the molecular defects of wnt signaling pathway and to treat cancers with such defects. PMID- 12642684 TI - Ribavirin in cancer immunotherapies: controlling nitric oxide helps generate cytotoxic lymphocyte. AB - Either ribavirin, RBV, or cyclophosphamide, CY, can shift an immune response from Th2 towards a Th1 cytokine profile. CY is used in this role in various current cancer immunotherapy attempts but with mixed success. More potent and reliable immunoadjuvants and Th 1 response biasing methods are needed. RBV is used today mainly to augment interferon-alpha treatment of hepatitis C. RBV shifts an immune response from Th2 towards Th1 more effectively than CY and may be a safe and useful adjuvant for current cancer immunotherapeutic efforts. RBV is thought to act by inhibition of tetrahydrobiopterin synthesis. Tetrahydrobiopterin is an essential co-factor for all known isoforms of nitric oxide synthase. Lowered nitric oxide favors Th1 development as high levels favor Th2 weighting. PMID- 12642685 TI - TRAIL-DISC formation is androgen-dependent in the human prostatic carcinoma cell line LNCaP. AB - We and others have previously described that the androgen-responsive human prostatic carcinoma cell line LNCaP is resistant to TRAIL and that TRAIL-mediated apoptosis in LNCaP is PI3K/Akt-dependent. In this study, we found that LNCaP remained resistant to treatment with TRAIL after androgen deprivation even in the presence of the PI3K/Akt pathway inhibitor wortmannin. This resistance was determined by failure to form the TRAIL-DISC and by decreased TRAIL-R1 and TRAIL R2 levels after androgen deprivation; the capacity of TRAIL to induce DISC formation was completely restored in the presence of DHT. TRAIL and wortmannin together accelerated processing of caspase-8 on the DISC and apparently the release of caspase-8 from the DISC into the cytoplasm. Surprisingly, we found that wortmannin decreased the total amount of TRAIL-R1, but not TRAIL-R2, in the cells as well as the amount of TRAIL-R1 precipitated by TRAIL. Our data suggest that TRAIL-DISC formation and sensitivity to TRAIL treatment are androgen dependent in LNCaP. PMID- 12642686 TI - On the TRAIL from death receptors to prostate cancer therapy. PMID- 12642687 TI - Developmental expression of Wnt signaling factors in mouse brain. AB - The Wnt signaling pathway has been implicated in a variety of biological events inducing neurogenesis. In this study, we aim to investigate the expression pattern of various components of the Wnt pathway including b-catenin and its partners LEF-1/TCF-4, GSK-3beta and their nuclear target genes such as c-myc and cyclin D1 during mouse brain development. We performed a series of Western blot and immunohistochemistry of brain cortex, brainstem, and cerebellum which revealed differential accumulation of these proteins in different types of brain cells including neurons, astrocytes, and oligodendrocytes at different developmental stages. Intense cytoplasmic immunolabeling of beta-catenin in 5 day old neurons throughout the cortex and brainstem significantly decreased as the brain developed, whereas the level of GSK-3beta, the protein that phosphorylates beta-catenin and causes its destabilization, increased during brain maturation. On the other hand, high level accumulation of LEF-1 and TCF-4 in neurons and astrocytes at the early stage of brain development diminished at the later stages. Interestingly, while the majority of LEF-1 and TCF-4 immunoreactivity was detected in the cytoplasm of neurons, it was evident that both proteins accumulated in the nuclei of astrocytes. Examination of cyclin D1, a protein that controls the cell cycle and proliferation, exhibited an intense staining in the nuclei of astrocytes throughout brain parenchyma during development. Interestingly, cyclin D was found in the cytoplasm of neurons from cortex, brainstem, and cerebellum during brain development. These data provide compelling evidence for the differential expression of the Wnt signaling pathway during brain development, and suggest that these signaling pathways may function differently in various brain regions and cell types. PMID- 12642688 TI - Phase I dose escalation trial of weekly docetaxel plus irinotecan in patients with advanced cancer. AB - Background. Docetaxel and irinotecan have additive or synergistic activity in vitro and in vivo as well as differing toxicities and unique mechanisms of action. We conducted a phase I trial to determine the maximum-tolerated dose of docetaxel and irinotecan given on a weekly schedule. Methods. Eligible patients had advanced, incurable, solid tumors. Docetaxel was administered as a 1-hour infusion and escalated over four dose levels (25, 30, 35, and 40 mg/m(2)) followed by irinotecan administered over 30 minutes at a fixed dose of 50 mg/m(2). Treatment was administered weekly for four weeks followed by two weeks of rest. To improve tolerability, the schedule was modified to weekly administration for two weeks with one week of rest, and irinotecan was escalated over 3 dose levels (55, 60, and 65 mg/m(2)) with docetaxel fixed at 35 mg/m(2). Results. Forty-four patients were treated and the most common dose-limiting toxicity was diarrhea observed in 11% of patients. Severe neutropenia was rare (grade 4: 2%, grade 3: 23%). Other nonhematologic toxicities included nausea/vomiting, dehydration and fatigue. Partial responses occurred in two patients with pancreatic cancer, and one patient each with non-small cell lung and esophageal cancer. Conclusions. Weekly docetaxel and irinotecan is a promising non-cisplatin doublet with preliminary evidence of activity in advanced solid tumors. Diarrhea is the predominant dose-limiting toxicity but unlike the every 3 weeks schedule myelosuppression is modest. The recommended phase II doses are docetaxel 35 mg/m(2) and irinotecan 60 mg/m(2) on days 1 and 8 of a 21-day schedule. Phase II trials of this regimen are ongoing or planned in lung, head and neck, stomach, esophageal, and pancreatic cancers. PMID- 12642690 TI - The clinical significance of tumor cell-lined vasculature in ovarian carcinoma: implications for anti-vasculogenic therapy. AB - Vasculogenic mimicry reflects the plasticity of aggressive tumor cells that express vascular cell markers and line tumor vasculature; such has been demonstrated in aggressive ovarian carcinoma. This study measured the clinical significance of tumor cell-lined vasculature in ovarian carcinomas (n=77), which was detected in 23 (29.8%) tumors. The data show that tumor cell-lined vasculature was associated with aggressive tumor features and with shorter overall survival (p<0.001). Cox proportional hazards model revealed that tumor cell-lined vasculature (p=0.002) was independently associated with poor survival. This is the first study demonstrating the clinical implications of tumor cell lined vasculature in ovarian carcinoma. PMID- 12642689 TI - Role of MRIT/cFLIP in protection against chemotherapy-induced apoptosis. AB - MRIT (Mach-related inducer of toxicity)/cFLIP (cellular FADD like interlukin1 beta converting enzyme inhibitory protein) is a proteolytically inactive structural homologue of caspase-8, which is known to protect against death receptors-induced apoptosis by blocking the activation of caspase-8. We have observed that exogenous expression of MRITalpha1/cFLIP(L) isoform also protects against cell death induced by a diverse group of chemotherapeutic drugs with different mechanisms of action, including doxorubicin, etoposide, cytosine arabinoside, daunorubicin, chlorambucil and cisplatin. However, MRITalpha1/cFLIP(L) failed to protect against apoptosis induced by paclitaxel and vincristine, two microtubule-damaging agents. Although MRITalpha1/cFLIP(L) protects against chemotherapy-induced apoptosis in both solid tumor and hematopoietic cell lines, this effect was more pronounced in the former. MRITalpha1/cFLIP(L) expression is decreased during drug-induced apoptosis and exogenous expression of MRITalpha1/cFLIP(L) delays the activation of caspase-8 and -3 during drug- induced apoptosis. These results suggest that MRITalpha1/cFLIPL may be an important determinant of both death receptor- and chemotherapy-induced apoptosis and strategies aimed at downregulating its expression deserve further study as a way to overcome multidrug resistance to cancer therapy. PMID- 12642691 TI - The histone deacetylase inhibitor FR901228 (desipeptide) restores expression and function of pseudo-null p53. AB - We have previously described a novel mechanism of p53 dysfunction, characterized by repression of mRNA and protein expression effectively leading to functional inactivation of wt p53 in SW-1736 human anaplastic thyroid cancer cells (pseudo null p53). Here we demonstrated that treatment of SW-1736 cells with sub cytotoxic concentrations of FR901228, a histone deacetylase (HDAC) inhibitor, results in marked induction of p53 mRNA and protein. The p53 induced by FR901228 was functional as evidenced by mdm-2 and p21 transactivation, and its further accumulation following DNA damage by doxorubicin. Furthermore, pretreatment with FR901228 sensitized SW-1736 cells to doxorubicin. This study validates the concept of pseudo-null p53, as a mechanism of p53 inactivation, and demonstrates that pseudo-null p53 can be rescued pharmacologically. PMID- 12642692 TI - Pharmacogenetic associations of CYP2C19 genotype with in vivo metabolisms and pharmacological effects of thalidomide. AB - Thalidomide requires cytochrome P450 (CYP)-catalyzed biotransformation for its antiangiogenic property, and CYP2C19 is responsible for 5-hydroxylation and 5' hydroxylation of thalidomide in human. This study explored a hypothesis that patients with poor metabolizing phenotype of CYP2C19 receive little benefit from thalidomide treatment and that the poor metabolizer genotype is associated with lower ability to form the metabolites. A case-control study was conducted with 63 patients with prostate cancer who had been enrolled in a randomized phase II trial of thalidomide monotherapy (200 to 1,200 mg/day). CYP2C19 polymorphism (CYP2C19(*)2, CYP2C19(*)3, CYP2C19(*)4) was compared with clinical events (prostate-specific antigen (PSA) decline) and formations of the hydroxylated metabolites. Two patients were homozygous for the variant CYP2C19(*)2 allele (poor metabolizing phenotype). Both of these were included in the 25 patients whose PSA failed to demonstrate a decline. While 32% and 48% of the patients had quantifiable levels of 5-hydroxythalidomide and cis-5'-hydroxythalidomide, respectively, these metabolite were below quantification in both poor metabolizing patients. None had CYP2C19(*)3 or CYP2C19(*)4 alleles. Although this study had no power to detect the statistical significance of the CYP2C19 genotype, the findings were consistent with our hypothesis. The role of CYP2C19 polymorphism in thalidomide treatments remains to be elucidated. PMID- 12642693 TI - Coadministration of UCN-01 with MEK1/2 inhibitors potently induces apoptosis in BCR/ABL+ leukemia cells sensitive and resistant to ST1571. AB - Interactions between the PKC and Chk1 inhibitor UCN-01 and pharmacologic MEK1/2 inhibitors (e.g., U0126, PD184352) were examined in Bcr/Abl(+) = human leukemia cells (K562, LAMA 84) sensitive and resistant to the Bcr/Abl kinase inhibitor STI571. Coexposure of K562 cells to UCN-01 (e.g., 100 nM) or U0126 (30 microM) resulted in a marked increase in mitochondrial injury (e.g., release of cytochrome c; loss of deltapsi(m)) and apoptosis. Similar results were obtained in other Bcr/Abl(+) cells (e.g., LAMA 84, BV-173) and with other MEK1/2 inhibitors (e.g., PD184352). Exposure of K562 cells to UCN-01 resulted in activation of ERK, an effect that was abrogated by co-administration of MEK1/2 inhibitors. Coadminstration of UCN-01 with U0126 produced multiple perturbations in signal transduction/cell cycle regulatory pathways, including diminished expression of Bcr/Abl, Mcl-1, cylin D(1), and activation of JNK and p34(cdc2). Coadministration of the JNK inhibitor SP600125 attenuated UCN-01/MEK inhibitor- associated lethality, suggesting a functional role for JNK activation in enhanced lethality. Finally, UCN-01 and MEK1/2 inhibitors effectively induced apoptosis in Bcr/Abl(+) cells (e.g., K562 and LAMA 84) overexpressing Bcr/Abl and resistant to STI571. These findings indicate that BcrAbl(+) leukemia cells are sensitive to a strategy combining UCN-01 with MEK/ERK inhibitors that simultaneously disrupts two signaling pathways. PMID- 12642694 TI - Beyond directed therapeutics--are two drugs always better than one? PMID- 12642695 TI - Dynamics of genetic instability in sporadic and familial colorectal cancer. AB - Genetic instability is a defining feature of human cancer. In colorectal cancer, two specific types of genetic instabilities have been identified: microsatellite instability (MIN) leads to a 1000-fold increase in the rate of subtle DNA changes, whereas chromosomal instability (CIN) enhances the rate at which gross chromosomal changes occur during cell division. In this paper, we develop a mathematical model for the dynamics of colon cancer initiation. We outline the processes and rate constants that determine the fraction of colon cancers where MIN or CIN mutations precede the inactivation of the first tumor suppressor gene. For a wide range of parameter values, we find support for the radical hypothesis that genetic instability initiates colonic tumorigenesis. We compare sporadic and hereditary forms of colorectal cancer. PMID- 12642696 TI - Single-nucleotide polymorphism (SNP) analysis in the ABC half-transporter ABCG2 (MXR/BCRP/ABCP1). AB - Variations in the amino acid sequence of ABC transporters have been shown to impact substrate specificity. We identified two acquired mutations in ABCG2, the ABC half-transporter overexpressed in mitoxantrone-resistant cell lines. These mutations confer differences in substrate specificity and suggest that naturally occurring variants could also affect substrate specificity. To search for the existence of single nucleotide polymorphisms (SNPs) in ABCG2, we sequenced 90 ethnically diverse DNAs from the Single Nucleotide Polymorphism Discovery Resource representing the spectrum of human genotypes. We identified 3 noncoding SNPs in the untranslated regions, 3 nonsynonymous and 2 synonymous SNPs in the coding region and 7 SNPs in the intron sequences adjacent to the sixteen ABCG2 exons. Nonsynonymous SNPs at nucleotide 238 (V12M; exon 2) and nucleotide 625 (Q141K; exon 5) showed a greater frequency of heterozygosity (22.2% and 10%) than the SNP at 2062 (D620N; exon 16). Heterozygous changes at nucleotide 238 are in linkage disequilibrium with an SNP observed 36 bases downstream from the end of exon 2. No polymorphism at amino acid 482 was identified to correspond to the R to G or R to T mutations previously found in two drug resistant cell lines. Among 23 drug resistant sublines for which sequence at position 482 was determined, no additional mutations were found. Heterozygosity at amino acid 12 allowed us to identify overexpression of a single allele in a subset of drug resistant cell lines, a feature that could be exploited clinically in evaluating the significance of ABCG2 expression in malignancy. We conclude that ABCG2 is well conserved and that described amino acid polymorphisms seem unlikely to alter transporter stability or function. PMID- 12642697 TI - Lyn tyrosine kinase inhibits nuclear export of the p53 tumor suppressor. AB - The p53 tumor suppressor is activated in the cellular response to stress. Mdm2 inhibits p53-dependent transactivation and promotes degradation of p53 by the ubiquitin-proteosome pathway. The present studies demonstrate that p53 binds directly to the nuclear Lyn tyrosine kinase. Lyn increases p53 levels and stimulates p53-mediated transcription by a kinase-independent mechanism. The results also demonstrate that Lyn increases nuclear levels of ubiquitinated p53 by inhibiting export of p53 to the cytoplasm. In concert with these results, Lyn reverses Mdm2-mediated degradation of p53 and increases p53-dependent apoptosis. Our findings support a previously undefined role for nuclear Lyn in both activation and Mdm2-mediated regulation of p53. PMID- 12642698 TI - Lyn: one more actor in the p53 cast? PMID- 12642699 TI - From the war on cancer to translational oncology. PMID- 12642700 TI - Fifty years in cancer research. PMID- 12642701 TI - Translational research: stop, look, listen--if you expect to grow up tall. PMID- 12642703 TI - LeukoScan for imaging infection in different clinical settings: a retrospective evaluation and extended review of the literature. AB - PURPOSE: The aim of the current study was to determine the overall diagnostic accuracy of Tc-99m-labeled antigranulocyte monoclonal antibody Fab' fragments (LeukoScan) for the routine detection of bone and soft tissue infections in a retrospective evaluation. PATIENTS AND METHODS: 138 patients (63 men, 75 women; mean age, 58.29 +/- 25.38 years) with fever of unknown origin and possible endocarditis (n = 59), infection of arthroplastic joints (n = 20), arthritis (n = 16), peripheral (n = 15) and central bone infections (n = 14), soft tissue infection (n = 6), appendicitis (n = 4), pericarditis (n = 2), or vascular graft infection (n = 2) underwent imaging after injection of 555 to 925 MBq (15 to 25 mCi) Tc-99m-labeled antigranulocyte monoclonal antibody Fab' fragments (LeukoScan). RESULTS: True-positive results were found in 63 of 81 lesions. The overall sensitivity and specificity were 76% and 84%, respectively. In arthritis, seven of seven foci could be detected, whereas false-negative results were found in infections of the femoral bone in three of nine lesions and in periprosthetic infections of long bones in three of eight lesions. Good results were found in five of six soft-tissue infections, in four of six patients with endocarditis, in three of four atypical cases of appendicitis, in two of two infected vascular grafts, and in one of one patient with pericarditis. Subacute and chronic infections of the spine always showed photopenic areas in eight of eight patients. If photopenic lesions were included as diagnostic criteria, the sensitivity and specificity were 88% and 67%, respectively. CONCLUSIONS: Tc-99m labeled antigranulocyte monoclonal antibody Fab' fragments can be used for imaging acute infections of peripheral bones and soft tissues. False-negative results are likely in patients with chronic infections. Sensitivity can be increased while decreasing specificity by including photopenic lesions in the spine as diagnostic criteria for localizing disease. PMID- 12642704 TI - Comparison of Tc-99m HIG and three-phase Tc-99m MDP bone scintigraphy for evaluating the efficacy of Yttrium-90 silicate radionuclide synovectomy. AB - PURPOSE: The aim of this study was to compare Tc-99m human immunoglobulin (HIG) and three-phase Tc-99m MDP bone scintigraphy for the assessment of the efficacy of Y-90 silicate therapy in rheumatoid knee synovitis. MATERIALS AND METHODS: Fifteen patients with rheumatoid arthritis and chronic persistent synovitis in 23 knee joints had radionuclide synovectomy with Y-90 silicate. The patients underwent imaging before and 3, 6, 9, and 12 months after therapy using clinical evaluation, Tc-99m HIG scintigraphy, and three-phase Tc-99m MDP bone scintigraphy. RESULTS: In the 13 of 23 knee joints that showed successful clinical results with Y-90 therapy, the Tc-99m HIG index values obtained 3 months after radionuclide synovectomy were significantly lower than the pretreatment index values (P < 0.001). In the same 13 joints, the Tc-99m MDP index values (in the blood-pool and delayed phases) before and 3 months after therapy were statistically similar. Six months after injection, these values were significantly lower in both the blood-pool (P < 0.001) and late (P < 0.05) phases in all 13 joints. In the other 10 of 23 knee joints that did not respond to treatment, the Tc-99m MDP and Tc-99m HIG index values were statistically similar before and after Y-90 therapy. CONCLUSIONS: Based on these findings, Tc-99m HIG scintigraphy appears to be a valuable method that complements clinical assessment of the efficacy of Y-90 silicate therapy in rheumatoid knee synovitis, starting in the early post-treatment period. However, three-phase Tc-99m MDP bone scintigraphy may be valuable in the late postsynovectomy period. PMID- 12642705 TI - Multifocal Pott's disease (tuberculous spondylitis) incidentally detected on Tc 99m MDP bone and Ga-67 citrate scintigraphy in a patient with diabetes. AB - Pott's disease is an uncommon extrapulmonary form of tuberculosis. Delay in diagnosis and management may cause serious complications. The authors describe Pott's disease incidentally detected on Tc-99m MDP bone and Ga-67 imaging in a patient with diabetes. Tc-99m MDP bone scintigraphy showed intensely increased uptake in the lower cervical spine and lumbosacral regions. Ga-67 scintigraphy revealed intensely increased uptake corresponding to the areas noted on Tc-99m MDP bone scintigraphy. Magnetic resonance imaging showed destructive lesions in the C5-C6 and L5-S1 intervertebral discs with destruction of adjacent end plates. Biopsy of the lumbosacral area was guided by computed tomography, and histologic examination of the bone specimen showed caseation, giant cells, and acid-fast bacilli. Posterior decompression and posterolateral spinal fusion with bone grafts were performed. Antituberculous chemotherapy with isoniazid, rifampicin, pyrazinamide, and ethambutol was started. The patient showed remarkable relief of symptoms during a period of 9 months of therapy. Both Tc-99m MDP bone and Ga-67 imaging can offer the convenience of screening the entire body to detect multiple sites of Pott's disease. PMID- 12642706 TI - Factors affecting left ventricular ejection fraction using automated quantitative gated SPECT. AB - BACKGROUND: Factors affecting the accuracy of left ventricular ejection fraction (LVEF) quantification using automated quantitative gated SPECT have not been adequately investigated in patients in the clinical setting. Therefore, the authors studied the effect of defect size and Tc-99m tetrofosmin dose on the accuracy of LVEF calculation using the automated QGS program. MATERIALS AND METHODS: Thirty-two consecutive patients underwent gated rest and stress myocardial perfusion SPECT after administration of 8 and 27 mCi Tc-99m tetrofosmin, respectively. The LVEF was obtained for both the rest and stress studies using the QGS program and compared with the LVEF obtained using quantitative echocardiography performed within 2 weeks. Myocardial perfusion defects were recorded as scarring, ischemia, or mixed scarring and ischemia in 12 left ventricular segments. The defect size was evaluated by adding the number of affected segments. RESULTS: The mean LVEF calculated using high-dose stress QGS, low-dose rest QGS, and echocardiography was 49.2% +/- 15%, 46.2% +/- 17% and 48.7% +/- 16.9% respectively, with no statistically significant differences. The LVEF obtained using high-dose stress QGS correlated better with echocardiography than did that obtained using low-dose rest QGS (r = 0.86 versus 0.76). In addition, when the high-dose stress LVEF in the 14 patients with normal myocardial perfusion was compared with that in 11 patients who had one- or two segment perfusion defects, and 7 patients who had perfusion defects in > or = three segments, there was good correlation with echocardiography in the three patient groups (r = 0.85, 0.88, and 0.91, respectively). CONCLUSIONS: Myocardial perfusion defects do not affect the accuracy of LVEF calculation using automated QGS. High-dose gated myocardial SPECT demonstrated better correlation with quantitative echocardiography LVEF results. PMID- 12642707 TI - Pituitary adenoma detected on FDG positron emission tomography in a patient with mucosa-associated lymphoid tissue lymphoma. AB - A 52-year-old woman with a history of treated non-Hodgkin's lymphoma, B-cell type, was examined for a right eyelid mass proved on excision to be a mucosa associated lymphoid tissue lymphoma. She was referred for F-18 fluorodeoxyglucose (FDG) positron emission tomography (PET) imaging of the whole body, including the brain, to identify the possible presence or recurrence of lymphoma. The FDG-PET images revealed focal marked FDG uptake in the pituitary gland but no evidence of abnormal metabolic sites in the rest of the body, including the orbital regions. Magnetic resonance imaging and computed tomography confirmed a 1.8 x 1.5 cm intrasellar mass. Surgery was performed, and histologic examination revealed a nonfunctional adenoma that was negative for leukocyte common antigen staining, an antigen that is positive in lymphoma but negative in pituitary adenoma. This case provides evidence for avid uptake of FDG on PET imaging of pituitary adenomas. PMID- 12642708 TI - False-negative Tc-99m MIBI scintigraphy in histopathologically proved recurrent high-grade oligodendroglioma. AB - Tc-99m MIBI is a small lipophilic radioligand that enters cells by diffusion and is preferentially trapped in mitochondria. As a result of the high mitochondrial activity in tumors, Tc-99m MIBI accumulates significantly more in tumor tissue compared with normal tissues. Accordingly, Tc-99m MIBI has been used successfully to visualize primary, metastatic, and recurrent tumor. In brain tumors, Tc-99m MIBI SPECT has been shown to identify tumor recurrence after treatment in high grade gliomas. In this report, early (30 minutes after injection) and delayed (4 hours after injection) Tc-99m MIBI SPECT did not visualize a histopathologically proved recurrent high-grade oligodendroglioma. Increased vascular supply, disruption of the blood-brain barrier, high-grade cancer, and viability of tumor cells are decisive factors related to increased Tc-99m MIBI uptake in brain tumors. However, the authors' results suggest that still other mechanisms may be involved in Tc-99m MIBI accumulation, which may account for false-negative imaging in brain tumors. PMID- 12642709 TI - Prevalence and patterns of bone metastases detected with positron emission tomography using F-18 FDG. AB - PURPOSE: The aim of this retrospective study was to report the prevalence and imaging characteristics of bone metastases detected with F-18 fluorodeoxyglucose (FDG) positron emission tomography (PET) and, when possible, compare these findings with the performance of bone scans in the same patients. METHODS: The reports of 403 patients with histologically proved malignant disease who underwent a PET scan for initial or post-therapeutic staging were reviewed for the presence of possible bone metastases. Based on the final diagnosis confirmed by histopathologic analysis or clinical follow-up, the PET findings of patients with positive bone metastases were evaluated in terms of location, intensity, and patterns. When the PET scan was positive, the PET results were compared with the findings of available bone scans. RESULTS: PET studies suggested the presence of bone metastases in 38 patients (9%). No follow-up data were available for 9 patients, and the remaining 29 were evaluated further. Of these patients, 6 had false-positive findings, whereas bone metastatic involvement was clinically confirmed in 23 patients. The primary malignant findings included lung cancer (n = 9), esophageal cancer (n = 3), lymphoma (n = 2), melanoma (n = 2), thyroid cancer (n = 2), breast cancer (n = 1), colon cancer (n = 1), prostate cancer (n = 1), testicular cancer (n = 1), and nasopharyngeal cancer (n = 1). On PET, 5 patients had a solitary metastatic focus (22%), and the remaining 18 patients had multiple lesions (78%). The vertebrae were the most frequently involved bones (74%), followed by pelvic bones (70%), ribs (65%), upper extremities including the scapula (48%), sternum (43%), and lower extremities (43%). The patterns of abnormal uptake were classified into three groups: focal (15 patients, 65%), diffuse (2 patients, 9%), and a mixed pattern (6 patients, 26%). Most of the lesions showed intense abnormal uptake (18 patients, 78%); 5 patients had both intense and moderate FDG uptake. Thirteen of the 23 patients with confirmed bone metastases also had a bone scan, which revealed positive bone disease in all of these patients. However, PET consistently revealed more metastatic foci than did the bone scan on a lesion basis. CONCLUSIONS: The most frequent pattern of detectable bone metastases with FDG-PET imaging was multiple foci of intense uptake. PET revealed more lesions than did bone scanning, independent of the type of cancer or location of bone involvement, in patients who were accurately diagnosed by FDG-PET imaging. PMID- 12642710 TI - Effects of menopause on bone mineral density in women with endemic fluorosis. AB - PURPOSE: The effects of menopause on bone mineral density (BMD) in women with endemic fluorosis were investigated. MATERIALS AND METHODS: Eighty healthy Turkish women who lived in and around the city of Isparta were selected randomly and enrolled in this study. They were separated into four groups: group 1, 20 premenopausal women with regular menstrual cycles and endemic fluorosis; group 2, 20 postmenopausal women with endemic fluorosis; group 3, 20 premenopausal normal women constituting one control group; and group 4, 20 postmenopausal normal women constituting the other control group. Bone mineral density was measured in the lumbar spine and proximal femur using dual-energy x-ray absorptiometry. RESULTS: In the premenopausal group, BMD values of vertebrae L2-L4 and Ward's triangle in women with endemic fluorosis were significantly greater than the respective values in women without endemic fluorosis (P = 0.024, P = 0.036). There were no differences between the groups in BMD values of the femoral neck (P = 0.156) and intertrochanteric area (P = 0.076). The BMD values of vertebrae L2-L4, the femoral neck, intertrochanteric area, and Ward's triangle in the postmenopausal women with endemic fluorosis were significantly greater than those of postmenopausal women without endemic fluorosis (P < 0.001, P = 0.015, P = 0.002, and P < 0.001, respectively). The BMD values of vertebrae L2-L4, the femoral neck, intertrochanteric area, and Ward's triangle in the premenopausal women with endemic fluorosis were significantly greater than those of postmenopausal women with endemic fluorosis (P = 0.010, P = 0.002, P = 0.004, and P = 0.010, respectively). The BMD values of the sites noted for the premenopausal controls were significantly greater than those of postmenopausal controls (P < 0.001, P < 0.001, P < 0.001, and P < 0.001, respectively). CONCLUSIONS: Postmenopausal BMD values in both endemic fluorosis and controls were significantly less than premenopausal BMD values. Although the differences were less prominent in women with endemic fluorosis, menopause is still the major determinant of BMD in the spine and femur. PMID- 12642712 TI - Detection of duplex kidney on a whole-body F-18 FDG positron emission tomographic scan. PMID- 12642711 TI - Native kidney recovery after a failed renal transplant. PMID- 12642713 TI - An endocrine active insulinoma with extended lymphogenic dissemination. PMID- 12642715 TI - Cervical lymphadenopathy caused by Kikuchi disease: positron emission tomographic appearance. PMID- 12642714 TI - Biliary tract FDG uptake secondary to stent placement. PMID- 12642716 TI - Discordant uptake on Ga-67 and Tc-99m MDP scintigraphy in a patient with multiple myeloma: an indication of poor prognosis. PMID- 12642717 TI - FDG positron emission tomographic imaging of thoracic Castleman's disease. PMID- 12642719 TI - Gastrointestinal bleeding, munchausen style. PMID- 12642718 TI - An FDG positron emission tomographic study in a case of gastrointestinal stromal tumor. PMID- 12642720 TI - Scintigraphic diagnosis of pleuroperitoneal communication in cirrhosis secondary to polycythemia vera. PMID- 12642721 TI - Accumulation of Tc-99m HMDP in bony structures of immature teratoma in a child. PMID- 12642722 TI - Bone scan findings in acronecrosis from cryoglobulinemia. PMID- 12642723 TI - Bone tracer uptake in urinary bladder stones. PMID- 12642724 TI - Tl-201 brain SPECT in glomus jugulare tumor. PMID- 12642725 TI - In-111 octreotide scintigraphy in adult medulloblastoma. PMID- 12642726 TI - Mastoid bone as a single site of metastasis of neuroendocrine tumor. PMID- 12642727 TI - Hiatal hernia mimics centrally necrotic cancer in the lung on FDG positron emission tomographic imaging. PMID- 12642728 TI - Arterial injection artifact on F-18 FDG positron emission tomographic scan. PMID- 12642729 TI - Iodine-131 uptake in focal bronchiectasis mimicking metastatic thyroid cancer. PMID- 12642730 TI - Different scintigraphic patterns of testicular lesions on dual-phase tl-201 scan. PMID- 12642732 TI - Eradication of Helicobacter pylori in nonulcer dyspepsia: how much analysis do we need? PMID- 12642733 TI - Management of Dieulafoy's lesion in the endoscopic era. PMID- 12642734 TI - Enteral nutritional support of the patient with cancer: route and role. AB - Malnutrition is a common and significant problem in patients with cancer. Enteral nutrition support is an important therapy and is preferred over parenteral nutrition in the setting of a functional gastrointestinal tract. Familiarity with the indications for enteral support, the choice of an enteral access device, and the selection of an enteral formula are critical for the care of patients with cancer and malnutrition. Enteral nutrition has proven efficacy in patients receiving radiation to the head and neck, those with persistent dysphagia, and critically ill patients with impaired gastric emptying. Placement of feeding tubes through the nose or percutaneously provides a mechanism to deliver nutrients when proximal obstructions or oropharyngeal dysphagia prevent adequate oral intake. Direct access to the jejunum can be safely obtained endoscopically and is very useful in patients who require enteral nutrition support following a gastrectomy or esophagectomy or have impaired gastric emptying. Standard polymeric formulas are appropriate for most patients. Specialized formulas designed to enhance immune function may decrease infectious complications but do not improve survival. PMID- 12642735 TI - Complications of pregnancy and child development after cessation of treatment with 6-mercaptopurine for inflammatory bowel disease. AB - PURPOSE: 6-Mercaptopurine (6-MP) has proven efficacy in the therapy of inflammatory bowel disease. Its teratogenicity is demonstrated in animal studies when used at very high doses, whereas human data suggest that 6-MP at maintenance doses is safe. We report the outcome of 72 pregnancies in patients with inflammatory bowel disease who were previously treated with 6-MP with three different doses of 50, 75, and 100 mg/d, for a median duration of 18 months, along with long-term follow-up of the children. METHODS: We have compared the outcome of pregnancies and development of the offspring in the following two groups: group 1, patients with inflammatory bowel disease who conceived 6 months to 22 years after stopping 6-MP (median 72 months); and group 2, patients with inflammatory bowel disease who never received 6-MP prior to conception. All pregnancies were evaluated in terms of outcome: live full-term birth, premature delivery, stillbirth, spontaneous abortion, ectopic pregnancy, and therapeutic dilatation and curettage. Data on children were obtained regarding birth weight, congenital anomalies, and development. RESULTS: Group 1 included 72 pregnancies carried by 29 women. There were 51 live births (4 premature), 16 spontaneous abortions, 1 stillbirth, 2 therapeutic abortions due to abnormal amniocentesis, and 2 ectopic pregnancies. The total incidence of fetal loss was 29.2%. In group 2, 75 women had 140 pregnancies resulting in 120 live births (8 premature), 18 spontaneous abortions, and 2 stillbirths. There were no cases of ectopic pregnancies or abnormal amniocentesis. The total incidence of fetal loss was 14.3%. There was no increase in the incidence of developmental defects when the mothers had been treated with 6-MP prior to pregnancy. CONCLUSIONS: The incidence of fetal loss is higher in women with inflammatory bowel disease who had been previously treated with 6-MP compared with those who had not. Whether this was related to the older age at conception in 6-MP group, longer duration of disease, initially more severe disease, or use of 6-MP we cannot tell. PMID- 12642736 TI - Outcomes of endoscopic treatment of gastroduodenal Dieulafoy's lesion with rubber band ligation and thermal/injection therapy. AB - BACKGROUND: Dieulafoy's lesion is a rare but important cause of upper gastrointestinal bleeding. Current endoscopic methods used to treat Dieulafoy's lesion include injection, with or without thermal methods, and mechanical methods. The latter include variceal ligation and hemoclips. There are no studies comparing the outcomes of rubber band ligation and injection with or without thermal therapy. AIM: To report the outcomes of Dieulafoy's lesion treated endoscopically with rubber band ligation and injection with or without thermal therapy at a single institution. METHODS: Patients with the diagnosis of Dieulafoy's lesion treated endoscopically at the Carl T. Hayden VA Medical Center in Phoenix, between August 1994 and August 2002 were analyzed. Demographic data, mode of presentation, risk factors for gastrointestinal bleeding, hemodynamic parameters, blood transfusion requirements, endoscopic findings, details of endoscopic therapy, length of stay in ICU/hospital, complications, recurrence of bleeding, and mortality rates were collected and compared between those receiving endoscopic band ligation (EBL group) and those receiving injection with or without thermal therapy (non-EBL group). RESULTS: Twenty-three patients with Dieulafoy's lesion (14 in the EBL group and nine in the non-EBL group) were studied. All patients were men. The mean age, hemoglobin levels on admission, and the transfusion requirements before therapy were similar in both groups. Fourteen patients (eight in the EBL- and six in the non-EBL groups) presented with hematemesis and the remaining with melena. The majority of Dieulafoy's lesions (91.3%) were located in the stomach and two in the duodenum. Active bleeding at the time of endoscopy was seen in 61% of cases, and immediate hemostasis was achieved with either method in 100% of patients. Early rebleeding (within 72 hours of endoscopic therapy) occurred in only one patient treated with epinephrine plus heater probe therapy. The length of stay in ICU was longer in the non-EBL group (6.7 days) compared with the EBL group (1.8 days) (P = 0.2). There were six deaths (three in the non-EBL group and three in the EBL group) within 30 days of the index hospitalization. The causes of death included infection/sepsis (n = 3), complications of acute myocardial infarction (n = 2), and end-stage liver disease (n = 1). CONCLUSIONS: Endoscopic rubber band ligation is as effective as injection with or without thermal therapy in the treatment of Dieulafoy's lesion. PMID- 12642737 TI - Helicobacter pylori infection treatment of nonulcer dyspepsia: an analysis of meta-analyses. AB - BACKGROUND: Meta-analysis is often used to resolve uncertainty in magnitude of effect between studies. However, several meta-analyses investigating the association between Helicobacter pylori infection and nonulcer dyspepsia reported contradictory results. GOALS: To perform a systematic review of the conflicting data in meta-analyses examining the association between H. pylori infection and nonulcer dyspepsia. STUDY: Five meta-analyses were identified: three studied the relation between H. pylori treatment and nonulcer dyspepsia, one studied the relation between H. pylori eradication and nonulcer dyspepsia, and one reported both. Data on search strategy, study selection, study inclusion, data abstraction, reported results of included studies, and summarized effect estimates were collected. RESULTS: Overall, 29 studies were included in the five meta-analyses. Nine studies (31%) were evaluated in more than one meta-analysis. There was no disagreement between the meta-analyses concerning the interpretation of outcome per investigated study; the results from seven (24%) studies showed improvement of dyspeptic symptoms after H. pylori treatment or eradication. The odds ratio of symptom relief in the control groups relative to the treated groups varied between 0.30 (95% CI = 0.2-0.5) to 0.88 (95% CI = 0.7-1.2). The proportion of patients' symptom free due to H. pylori treatment varied between 11% and 38%. CONCLUSIONS: The reported conflicting results between the meta-analyses were mainly a result of differences in study selection (eligibility and quality criteria) and relatively small. In a subgroup of patients with nonulcer dyspepsia, H. pylori eradication is useful for relief of symptoms. PMID- 12642738 TI - Helicobacter pylori infection and early gastric cancer. AB - BACKGROUND: Helicobacter pylori has recently been associated with an increased risk of gastric cancer. This study aimed to examine the association between H. pylori, histologic chronic gastritis, and intestinal metaplasia in early gastric cancers of different histologic types. STUDY: Seventy-four patients who were surgically diagnosed as having early gastric cancer were included in this study. All tissue specimens were obtained from patients by endoscopic biopsy and were classified histopathologically as intestinal-type early gastric cancer in 55 patients and diffuse-type early gastric cancer in 19 patients. RESULTS: H. pylori infection was found in 67 patients (90.5%) but not found in seven (9.5%). And the prevalence of H. pylori infection with nongastric cancer patients was also high (68.5%). There was no significant difference between the intestinal-type and the diffuse-type early gastric cancer in chronic active gastritis and atrophic chronic gastritis. Intestinal metaplasia was observed more frequently in patients with the intestinal-type than with the diffuse-type early gastric cancer (P = 0.0102). CONCLUSIONS: Infection with H. pylori has an important relationship to both histopathologic types of early gastric cancer. PMID- 12642739 TI - Omeprazole plus azithromycin and either amoxicillin or tinidazole for eradication of Helicobacter pylori infection. AB - GOALS: To establish whether omeprazole plus azithromycin in association with either amoxicillin or tinidazole is effective in curing Helicobacter pylori infection in dyspeptic patients. BACKGROUND: Many antibiotics in combination with antisecretory drugs have been used in an attempt to find the optimal regimen for eradication of H. pylori. Azithromycin is a macrolide that achieves high concentrations in gastric tissue after a single 500-mg oral dose. STUDY: A total of 160 consecutive symptomatic patients with H. pylori received omeprazole 20 mg twice daily for 1 week, azithromycin 500 mg/d for 3 days, and were randomly assigned to either amoxicillin 1 g twice daily (OAzAm group, n = 80) for 1 week or tinidazole 500 mg twice daily for 3 days (OAzT group, n = 80). H. pylori status was assessed by rapid urease test and histology at entry and by histology and (13)C-urea breath test after the end of the therapy. RESULTS: H. pylori was eradicated in 62.5% of patients in the OAzAm group (intention to treat [ITT] 62.5%) and in 71.2% of patients in the OAzT group (ITT 71.2%). CONCLUSIONS: Although the compliance was excellent and the side effects negligible, the regimens used were partially effective for the eradication of H. pylori. PMID- 12642740 TI - Urinary antibody titers to Helicobacter pylori and an impact of clinical characteristics. AB - This study evaluates the clinical usefulness of titers of antibody to Helicobacter pylori (Hp) in a urine-based enzyme-linked immunosorbent assay (URINELISA; Otsuka Pharmaceutical, Tokushima, Japan) by comparing values of serum antibody to Hp in an already commercialized kit in a population of 250 asymptomatic individuals. Influences of physical and laboratory characteristics on the relationship between serum and urinary titers of antibody to Hp were also estimated using simple and multiple regression analyses. Specific urine gravity, urine pH, proteinuria, and age of the subjects were regarded as significant factors relating to urinary titers of the anti-Hp antibody on simple regression analyses. Both the urinary and serum titers strongly correlated on simple regression analysis (r = 0.782, P < 0.0001). On multiple regression analysis, an additional two factors, specific urine gravity and urine pH, were picked up, and the correlation coefficient was improved (r = 0.795, P < 0.0001). The present results indicate that URINELISA had not only qualitative but quantitative accuracy. However, careful attention should be paid to cases with abnormalities in urinalysis when used quantitatively. PMID- 12642741 TI - Primary intestinal lymphoma complicating Crohn's disease. AB - A 32-year-old man with a 12-year history of Crohn's disease of the colon was found to have a 5-cm cecal mass on colonoscopy. Histology examination of the lesion revealed high-grade B-cell lymphoma of Burkitt's type. He was treated with chemotherapy and has done well in the past 12 months. Review of the literature reveals 30 cases of lymphoma in patients with a history of Crohn's disease. PMID- 12642742 TI - Tropical chronic pancreatitis: an update. AB - Tropical chronic pancreatitis represents a juvenile nonalcoholic form of chronic pancreatitis prevalent in many tropical developing countries. Tropical chronic pancreatitis differs from temperate zone pancreatitis in its younger age of onset, more accelerated course, higher prevalence of pancreatic calculi and diabetes, and greater propensity to pancreatic malignancy. The diabetic stage of the disease is referred to as fibrocalculous pancreatic diabetes. The diabetes is severe and insulin requiring although ketosis resistant. Diabetic complications occur in fibrocalculous pancreatic diabetes just like in other primary forms of diabetes. The etiology of tropical chronic pancreatitis remains unclear, although malnutrition along with dietary cyanogen toxicity, antioxidant deficiency, and a genetic predisposition have been proposed. In the last few decades, the prognosis has markedly improved as a result of better management of diabetes. Yet a better understanding of the pathogenesis of tropical chronic pancreatitis could further improve treatment options and offer an opportunity for prevention of this disorder, which leads to severe morbidity in a large proportion of affected patients. PMID- 12642743 TI - Treatment of unresectable hepatocellular carcinoma less than 2 centimeters by transcatheter arterial chemoembolization with autologous blood clot. AB - GOALS: To assess the efficacy of transcatheter arterial chemoembolization using autologous blood clot as an embolizing agent (short-TAE [S-TAE]) for the treatment of unresectable hepatocellular carcinoma less than 2 cm. STUDY: Twenty eight consecutive patients with unresectable hepatocellular carcinoma less than 2 cm in diameter were treated by S-TAE alone. All patients had documented cirrhosis (Child class B:C = 20:8). S-TAE was performed by injecting a mixture of iodized oil and anticancer drugs followed by embolization of hepatic arteries with autologous blood clot. RESULTS: A total of 147 sessions of embolization with clots were performed. S-TAE maintained patency of hepatic arteries. The overall survival rates at 1, 3, 5, and 8 years were estimated to be 89%, 52%, 34%, and 17%, respectively, which were better compared with prior records for the gelfoam method. The survival rates for Child class B patients were significantly better than that for Child class C patients (P < 0.05). The Cox proportional hazard model also demonstrated that Child staging of cirrhosis was the sole factor significantly predicting the survival (P < 0.05). CONCLUSIONS: The long-term outcomes of S-TAE for unresectable hepatocellular carcinoma less than 2 cm are satisfactory. Prognosis of these patients was significantly dependent on clinical stages of coexisting liver cirrhosis. PMID- 12642744 TI - A pilot study of the combination of cyclosporin A and interferon alfacon-1 for the treatment of hepatitis C in previous nonresponder patients. AB - GOALS: To evaluate the safety and efficacy of the combination of interferon and cyclosporine for the treatment of hepatitis C in previous nonresponder patients. BACKGROUND: Preliminary data indicated that adding the immunosuppressive agent cyclosporin A to interferon might improve response rates in patients with hepatitis C. STUDY: Ten previous virologic nonresponders with genotype 1 infection were included. Treatment consisted of interferon alfacon-1, 15 microg/d, and cyclosporine, 100 mg twice daily, for 4 weeks. The dose of interferon alfacon-1 was then decreased to 15 microg three times weekly, and cyclosporine was reduced to 50 mg twice daily. Therapy was continued for 48 weeks unless viremia persisted at week 24. RESULTS: Three of 10 subjects had an on treatment virologic response, although one had a breakthrough with recurrent viremia during treatment and two relapsed after therapy was completed. On treatment responders had significantly higher trough cyclosporine levels at week 4 compared with nonresponders (P = 0.025). Serum creatinine levels remained stable, and no patient developed diabetes. Triglyceride levels increased during treatment. Cyclosporine was dose reduced in two patients for hypertension. CONCLUSIONS: Selected patients with hepatitis C tolerated therapy, including cyclosporine without severe or irreversible toxicity. Despite an association between higher cyclosporine levels and on-treatment response, the combination of cyclosporine and interferon was ineffective in producing a sustained response in previous nonresponder patients. PMID- 12642745 TI - Risk of cancer in patients hospitalized with fatty liver: a Danish cohort study. AB - GOALS: To examine the risk of cancer in patients with fatty liver. BACKGROUND: The relation between liver disease, including fatty liver, and cancer risk is poorly understood. STUDY: Using the population-based National Registry of Patients, we examined the incidence of cancer in 7,326 patients discharged with a diagnosis of fatty liver from a Danish hospital during 1977-1993. Patients with a prior diagnosis of liver cirrhosis were excluded. We identified cancers through the Danish Cancer Registry. The expected number of cancers was estimated from national age-, sex-, and site-specific incidence rates. RESULTS: Overall, 523 cancers were diagnosed during 47,594 person-years of follow-up, yielding a 1.7 fold increased risk (95% confidence interval, 1.6-1.9) compared with the Danish general population. The risk of primary liver cancer was markedly elevated in patients with alcoholic as well as nonalcoholic fatty liver with a standardized incidence ratio of 9.5 (95% confidence interval, 5.7-14.8) and 4.4 (95% confidence interval, 1.2-11.4), respectively. Patients with alcoholic fatty liver also had substantially increased risks of several types of cancer associated with alcohol and tobacco use (cancers of the lung, pharynx, larynx, esophagus, and stomach) and a moderately increased risk for cancers of the colon and breast. Among patients with nonalcoholic fatty liver, an increased risk of some alcohol- and tobacco-related cancers was seen, and there was also an increased risk of colon and pancreas cancer. CONCLUSIONS: Patients discharged with a diagnosis of fatty liver have an increased risk of cancer, in particular liver cancer, most prominently among patients with alcoholic fatty liver. PMID- 12642746 TI - Early activation of peripheral lymphocytes in human acute pancreatitis. AB - BACKGROUND: The CD69 antigen is an indicator of early lymphocyte activation. GOALS: To evaluate the early activation of peripheral lymphocytes T, B, and NK in patients with acute pancreatitis in comparison with patients with acute abdomen of nonpancreatic origin. STUDY: Thirty patients with acute pancreatitis were studied; 20 of them had the mild form of the disease and 10 had the severe form. Thirty patients with nonpancreatic acute abdomen were used as controls. All patients were enrolled within 48 hours of the onset of pain. In all patients, leukocytes and total lymphocyte and lymphocyte subset counts (CD4+, CD8+, CD56+, CD19+, CD4+CD69+, CD8+CD69+, CD56+CD69+, CD19+CD69+) were determined upon hospital admission. RESULTS: The percentage of total lymphocytes was significantly lower in acute pancreatitis patients than in those with nonpancreatic acute abdomen (P = 0.014); patients with severe pancreatitis had a percentage of total lymphocytes significantly lower when compared with patients with mild pancreatitis (P < 0.001). The CD19+CD69+ count was significantly lower in patients with severe pancreatitis (24.6 +/- 14.6%) than in patients with mild pancreatitis (46.7 +/- 16.5%; = 0.006). The counts of the other lymphocyte subsets were not statistically different between patients with acute pancreatitis and those with nonpancreatic acute abdomen, as well as between patients with mild and severe acute pancreatitis. CONCLUSIONS: Patients with severe pancreatitis show impaired early activation of peripheral CD19+ cells. PMID- 12642747 TI - Cholesterol crystal morphology in acalculous gallbladder disease. AB - BACKGROUND: Biliary crystal morphology is best described in patients with gallbladder stones, but most patients undergoing bile collection for microscopy have a clinical diagnosis of acalculous gallbladder disease. We investigated the morphology of biliary crystals in such patients. STUDY: Bile was obtained for polarizing microscopy from fresh cholecystectomy specimens of patients with a clinical diagnosis of acalculous or calculous gallbladder disease. Slides for microscopy were prepared by touch contact with bile in freshly opened gallbladder specimens, and following aspiration of gallbladder bile through a 5-French cannula. RESULTS: Bile was examined from five patients with a clinical diagnosis of acalculous gallbladder disease and five patients with known gallstones. Needle like cholesterol crystals predominated in most patients without gallstones, whereas plate-like and dot-like crystals were more common in patients with gallstones. All three crystal types were seen in most patients. Crystal morphology was not affected by aspiration of bile through a 5-French cannula. CONCLUSIONS: Birefringent needles and dots should be recognized as cholesterol crystals during bile microscopy. These crystal morphologies may predominate in some patients with a clinical diagnosis of acalculous gallbladder disease. PMID- 12642748 TI - Hemoglobinuria with ribavirin treatment. AB - Chronic hepatitis C virus is a major worldwide cause of hepatitis, cirrhosis, end stage liver disease, and hepatocellular carcinomas. Combination therapy of ribavirin with short- or long-acting interferon-alpha is now the standard treatment of chronic hepatitis C. This therapy is associated with a wide range of side effects. Although hemolysis is almost an invariable result of ribavirin, black urine due to hemoglobinuria has never been previously reported. We recently encountered two cases of black urine (hemoglobinuria) in patients treated with combination therapy. Based on reports of dark urine in many of our patients, we suggest that this phenomenon may be more common than is currently appreciated. It indicates a marked degree of hemolysis, which prompts immediate measurement of hemoglobin level. PMID- 12642749 TI - Increased incidence of fungal infections in advanced primary biliary cirrhosis. PMID- 12642750 TI - Multiple liver abscesses after dental treatment. PMID- 12642751 TI - The fluctuation of serum levels of aminotransferase in patients with nonalcoholic steatohepatitis. PMID- 12642752 TI - How should we improve the clinical course of patients with gastric varices? PMID- 12642753 TI - Successful treatment of acute hepatic injury caused by paracetamol intoxication in a late-referral patient by N-acetylcysteine. PMID- 12642754 TI - Metronidazole and mental confusion. PMID- 12642755 TI - Manual therapy and exercise therapy in patients with chronic low back pain: a randomized, controlled trial with 1-year follow-up. AB - STUDY DESIGN: A multicenter, randomized, controlled trial with 1-year follow-up. OBJECTIVES: To compare the effect of manual therapy to exercise therapy in sick listed patients with chronic low back pain (>8 wks). SUMMARY AND BACKGROUND DATA: The effect of exercise therapy and manual therapy on chronic low back pain with respect to pain, function, and sick leave have been investigated in a number of studies. The results are, however, conflicting. METHODS: Patients with chronic low back pain or radicular pain sick-listed for more than 8 weeks and less than 6 months were included. A total of 49 patients were randomized to either manual therapy (n = 27) or to exercise therapy (n = 22). Sixteen treatments were given over the course of 2 months. Pain intensity, functional disability (Oswestry disability index), general health (Dartmouth COOP function charts), and return to work were recorded before, immediately after, at 4 weeks, 6 months, and 12 months after the treatment period. Spinal range of motion (Schober test) was measured before and immediately after the treatment period only. RESULTS: Although significant improvements were observed in both groups, the manual therapy group showed significantly larger improvements than the exercise therapy group on all outcome variables throughout the entire experimental period. Immediately after the 2-month treatment period, 67% in the manual therapy and 27% in the exercise therapy group had returned to work (P < 0.01), a relative difference that was maintained throughout the follow-up period. CONCLUSIONS: Improvements were found in both intervention groups, but manual therapy showed significantly greater improvement than exercise therapy in patients with chronic low back pain. The effects were reflected on all outcome measures, both on short and long-term follow-up. PMID- 12642757 TI - Mini-intervention for subacute low back pain: a randomized controlled trial. AB - STUDY DESIGN: Randomized controlled trial. OBJECTIVES: To investigate the effectiveness and costs of a mini-intervention, provided in addition to the usual care, and the incremental effect of a work site visit for patients with subacute disabling low back pain. SUMMARY OF BACKGROUND DATA: There is lack of data on cost-effectiveness of brief interventions for patients with prolonged low back pain. METHODS: A total of 164 patients with subacute low back pain were randomized to a mini-intervention group (A), a work site visit group (B), or a usual care group (C). Groups A (n = 56) and B (n = 51) underwent one assessment by a physician plus a physiotherapist. Group B received a work site visit in addition. Group C served as controls (n = 57) and was treated in municipal primary health care. All patients received a leaflet on back pain. Pain, disability, specific and generic health-related quality of life, satisfaction with care, days on sick leave, and use and costs of health care consumption were measured at 3-, 6-, and 12-month follow-ups. RESULTS: During follow-up, fewer subjects had daily pain in Groups A and B than in Group C (Group A Group C, = 0.002; Group B Group C, = 0.030). In Group A, pain was less bothersome (Group A Group C, = 0.032) and interfered less with daily life (Group A Group C, = 0.040) than among controls. Average days on sick leave were 19 in Group A, 28 in Group B, and 41 in Group C (Group A Group C, = 0.019). Treatment satisfaction was better in the intervention groups than among the controls, and costs were lowest in the mini-intervention group. CONCLUSIONS: Mini-intervention reduced daily back pain symptoms and sickness absence, improved adaptation to pain and patient satisfaction among patients with subacute low back pain, without increasing health care costs. A work site visit did not increase effectiveness. PMID- 12642759 TI - The pylon concept of pelvic anchorage for spinal instrumentation in the human cadaver. AB - STUDY DESIGN: Human cadavera morphometric analysis of the iliac columns and biomechanical implant testing of traditional Galveston technique compared to intrailiac instrumentation of the entire iliac column. OBJECTIVES: To describe the anatomy of the iliac columns and to evaluate the strength in forward flexion of a large implant spanning the entire column length compared to standard Galveston technique. SUMMARY OF BACKGROUND DATA: We have observed substantial and straight columns of bone in the pelvis, connecting the acetabula to the sacrum, which may allow for improved spinopelvic instrumentation. METHODS: Twenty adult cadaveric pelves were used. Each specimen was oriented in the computed tomography scanner to obtain a cross-section of the iliac columns, which begin from 2 cm caudal to the posterior iliac spines and end above the acetabula at the anterior inferior iliac spines. Two different instrumentation techniques were used. Standard Galveston pelvic fixation with paired 6.25-mm diameter rods extending 8 cm into the pelvis (Group 1) was compared to paired 8-mm diameter, 15-cm long custom implants, placed within the length of the entire iliac columns and connected to 6.25-mm spinal rods (Group 2). Both constructs had two rigid cross links connecting the rods. Testing in forward flexion was performed for each construct with the MTS model 881 at 5 N/sec until failure occurred. RESULTS: The rectangular shaped iliac columns averaged 15.2 (SD 0.8) cm in length, 2.5 (SD 0.3) cm in width and were consistently straight. The iliac column orientation as viewed in the transverse plane was 22 degrees laterally directed from the midsagittal plane. For the Galveston technique, failure with a flexion force occurred at a mean of 682 (SD 217) N. The iliac column implants failed at a mean of 2153 (SD 1370) N (P < 0.004). CONCLUSION: The human adult pelvis has substantial and straight columns of bone extending from 2 cm below the posterior iliac spine, traversing above the sciatic notch, and ending at the anterior iliac spine. The shape resembles a weight-bearing long bone such as the tibia. Analogous to the architectural pylon, in this cadaver model, large implant instrumentation of the entire length of these pelvic columns provides at least three times stronger anchorage for spinal instrumentation compared to standard Galveston technique. PMID- 12642760 TI - An experimental study of the regeneration of the intervertebral disc with an allograft of cultured annulus fibrosus cells using a tissue-engineering method. AB - STUDY DESIGN: Cultured annulus fibrosus cells within an atelocollagen honeycomb shaped scaffold with a membrane seal were allografted into the lacunas of intervertebral discs of which the nucleus pulposus had been vaporized using an indocyanine green dye-enhanced laser. Regeneration of the intervertebral disc was assessed based on the viability and histologic status of the allografted annulus fibrosus cells, as well as the prevention of narrowing disc space. OBJECTIVES: To study the regeneration of intervertebral disc after laser discectomy using tissue engineering methods. SUMMARY OF BACKGROUND DATA: Intervertebral disc is the most avascular tissue in the human body, and its ability to regenerate is as low as that of articular cartilage. When nucleotomy is carried out, little regeneration of the annulus fibrosus is observed; consequently, intervertebral disc degeneration is inevitable. METHODS: Annulus fibrosus cells isolated from 20 Japanese white rabbits were labeled with a PKH-26 fluorescent dye and seeded within an atelocollagen honeycomb-shaped scaffold with a membrane seal. Annulus fibrosus cells cultured in atelocollagen honeycomb-shaped scaffold with a membrane seal for 1 week were allografted into the lacunas of intervertebral discs of recipient rabbit of which the nucleus pulposus had been vaporized using an ICG dye-enhanced laser. Soft radiograph photographs of the lumbar spine of these anesthetized rabbits were taken, the disc space measured, and the lumbar spine extracted 2, 4, 8, and 12 weeks after the operation. The proliferation of allografted annulus fibrosus cells with 5-bromo-2'-deoxyuridine/PKH-26 fluorescent labels was assessed using consecutive frozen sections, and safranin-O staining carried out for histologic evaluation. RESULTS: The allografted annulus fibrosus cells were viable and showed proliferation activity with a hyaline-like cartilage being produced. The narrowing of the intervertebral disc space of the cell translation group was significantly prevented, as shown, up to 12 postoperative weeks. CONCLUSION: The annulus fibrosus cells cultured in an ACHMS scaffold were allografted into the lacunae of nucleus pulposus (obtained using laser vaporization), as well as the hole of annulus fibrosus (obtained by laser fiber insertion) of rabbit intervertebral discs. These cells were viable and showed cell proliferation in the disc tissues of recipients. PMID- 12642761 TI - Implication for melatonin and its receptor in the spinal deformities of hereditary Lordoscoliotic Rabbits. AB - STUDY DESIGN: The association between melatonin system and the spontaneous development of the spinal deformities in the Hereditary Lordoscoliotic Rabbit, the natural animal model for idiopathic scoliosis, was studied. OBJECTIVES: To examine the implication for melatonin and its receptor in the spinal deformities of the natural animal model, the Hereditary Lordoscoliotic Rabbit. SUMMARY OF BACKGROUND DATA: We previously reported radiologic and histologic studies investigating the etiology of spinal deformities in a breed of Japanese White Rabbit, the Hereditary Lordoscoliotic Rabbit. These animals develop thoracic lordoscoliosis during growth and as such can be used as a model for human idiopathic scoliosis. Although previous studies in chickens have established that pinealectomy produces scoliosis, the cause of the condition is yet to be fully elucidated. METHODS: Serum melatonin levels in Hereditary Lordoscoliotic Rabbits were measured by radioimmunoassay and compared with those of Japanese White Rabbits (controls). The expression of melatonin receptor in the rabbit was detected by homology cloning to access the number of the melatonin receptor mRNA in the rabbit spinal cord by quantitative reverse-transcribed polymerase chain reaction. RESULTS: Serum melatonin levels in Hereditary Lordoscoliotic Rabbits were significant higher than those of controls in each period until 20 weeks. We detected the expression of melatonin receptor mRNA in rabbit spinal cord. However, no significant quantitative differences were found in the level of expression of melatonin mRNA in the spinal cord between Hereditary Lordoscoliotic Rabbits and controls. CONCLUSIONS: In relation to the present study, we suggest that causes of spinal deformities in the Hereditary Lordoscoliotic Rabbit may be the result of the contribution of melatonin receptors as well as that of altered serum melatonin levels in the Hereditary Lordoscoliotic Rabbit. Further studies will be required to investigate the expression of melatonin receptor in other tissues of the Hereditary Lordoscoliotic Rabbit as well as to delineate the role of melatonin in the pathogenesis of idiopathic scoliosis. PMID- 12642762 TI - Finite element modeling of the human thoracolumbar spine. AB - STUDY DESIGN: Biomechanical properties within cadaveric vertebral bodies were parametrically studied using finite element analysis after calibration to experimental data. OBJECTIVES: To develop and validate three-dimensional finite element models of the human thoracolumbar spine based on quantitative computed tomography scans. Specifically, combine finite element modeling together with biomechanical testing circumventing problems associated with direct measurements of shell properties. SUMMARY OF BACKGROUND DATA: Finite element methods can help to understand injury mechanisms and stress distribution patterns within vertebral bodies as an important part in clinical evaluation of spinal injuries. Because of complications in modeling the vertebral shell, it is not clear if quantitative computed tomography-based finite element models of the spine could accurately predict biomechanical properties. METHODS: We developed a novel finite element modeling technique based on quantitative computed tomography scans of 19 radiographically normal human vertebra bodies and mechanical property data from empirical studies on cylindrical trabecular bone specimens. Structural properties of the vertebral shell were recognized as parametric variables and were calibrated to provide agreement in whole vertebral body stiffness between model and experiment. The mean value of the shell properties thus obtained was used in all models to provide predictions of whole vertebral strength and stiffness. RESULTS: Calibration of n = 19 computer models to experimental stiffness yielded a mean effective modulus of the vertebral shell of 457 +/- 931 MPa ranging from 9 to 3216 MPa. No significant correlation was found between vertebral shell effective modulus and either the experimentally measured stiffness or the average trabecular modulus. Using the effective vertebral shell modulus for all 19 models, the predicted vertebral body stiffness was an excellent predictor of experimental measurements of both stiffness (r2= 0.81) and strength (r2 = 0.79). CONCLUSION: These findings indicate that modeling of the vertebral shell using a constant thickness of 0.35 mm and an effective modulus of 457 MPa, combined with quantitative computed tomography-based modeling of trabecular properties and vertebral geometry, can accurately predict whole vertebral biomechanical properties. Use of this modeling technique, therefore, should produce substantial insight into vertebral body biomechanical behavior and may ultimately improve clinical indications of fracture risk of this cohort. PMID- 12642763 TI - C1-C2 posterior fusion in growing patients: long-term follow-up. AB - STUDY DESIGN: A retrospective review of patients undergoing C1-C2 posterior fusion during childhood was undertaken. OBJECTIVES: The aim of this study was to investigate the change in the sagittal curvature of the cervical spine in children after C1-C2 posterior fusion. SUMMARY OF BACKGROUND DATA: There have been only a few reports on postoperative changes in the sagittal curvature of the cervical spine after C1-C2 posterior fusion in children. However, they have all described the onset of sagittal postoperative cervical deformities. METHODS: Between January 1977 and December 1992, a total of 12 children underwent C1-C2 posterior fusion for atlantoaxial instability resulting from congenital malformation in eight, juvenile rheumatoid arthritis in one, and rotatory subluxation in three. The average age at the time of surgery was 10.9 years (range 7-12 years). All children underwent a similar treatment program with gradual preoperative reduction in halo cast, followed by C1-C2 posterior fusion with Mersilene loops in two cases, wiring in eight (Gallie's or Brooks' techniques), and interlaminar clamps in the remaining two. The halo cast made it possible to avoid a hyperextended or hyperflexed C1-C2 position while performing the atlantoaxial fusion, thus ensuring a more anatomic position during C1-C2 fusion. In the postoperative period, the halo cast was maintained for 7 to 9 weeks. RESULTS Follow-up ranged from 7 years to 13 years. Preoperative alignment of the cervical spine was classified into two groups: lordosis (eight patients) and straight (four patients). Postoperative subaxial malalignment (kyphosis) occurred in four cases (33%): these patients showed evidence of spontaneous and gradual sagittal improvement and presented either a straight (two cases) or a lordotic (two cases) cervical spine at follow-up. Immediately after surgery, the cervical spine was normally aligned in the remaining eight patients (lordosis and straight alignment in six and two cases, respectively) and was unchanged at follow-up. At follow-up, none of the 12 patients had a cervical deformity on sagittal plane. CONCLUSION: In children, a spontaneous realignment of the subaxial kyphosis observed after C1-C2 posterior fusion can be noted at follow up, when a postoperative deformity occurs (33% in the present series). According to the present findings, it is not always mandatory to perform occipitocervical fusion in children with atlantoaxial instability just to prevent subaxial deformity in the cervical spine. PMID- 12642765 TI - Management of isthmic spondylolisthesis with posterolateral endoscopic foraminal decompression. AB - STUDY DESIGN: Prospective evaluation of 24 consecutive patients with isthmic spondylolisthesis with chronic back, buttock, and leg pain treated by endoscopic foraminal decompression and followed for a minimum of 2 years. OBJECTIVES: To assess the efficacy of endoscopic foraminal decompression and mobilization of the exiting and transiting nerves, discectomy, ablation of osteophytes, and impinging pars as a means of treatment by the posterolateral approach. SUMMARY OF BACKGROUND DATA: Open decompression with or without fusion is a commonly accepted procedure for symptomatic isthmic spondylolytic spondylolisthesis in patients who fail to respond to conservative treatment. There is no published data on the outcome of endoscopic procedures for this condition. METHODS: Endoscopic foraminal decompression achieved with laser-assisted bone and soft-tissue ablation was performed on 12 males and 12 females with an average age of 42.4 years (36-72 years) followed for an average period of 34 months (28-46 months). The average preoperative duration of symptoms was 6.1 years (3-9 years). RESULTS: One hundred percent cohort integrity was maintained at the final follow-up. Results were analyzed using the percentage change in Oswestry Disability Scores and in Visual Analogue Pain scores. Using a percentage change in Oswestry Disability Score of 50 or more plus VAP scores of 50 or more to determine good and excellent outcomes, 79% (19 out of 24) exceeded this value. CONCLUSION: Laser assisted endoscopic foraminal decompression provides a minimalist means of exploring the extraforaminal zone, the isthmic defect, the foramen and its contents, the disc and the epidural space. It allows adequate resection with decompression and discectomy, without the need for open decompression and fusion, and targets the symptomatic level effectively in patients with Grade I-III isthmic spondylolisthesis. PMID- 12642766 TI - Associations between back pain history and lumbar MRI findings. AB - STUDY DESIGN: Retrospective monozygotic twin cohort study. OBJECTIVES: Our goal was to investigate the associations between different spinal MRI findings and current, past year, and lifetime low back pain after adjusting for occupational physical loading, smoking, genetics, and early family influences. SUMMARY OF BACKGROUND DATA: The role of spinal pathology in back symptoms continues to be controversial. METHODS: The study participants consisted of 115 monozygotic male twin pairs 35 to 69 years of age. The qualitatively assessed MRI parameters were as follows: disc height, bulging, herniations, anular tears, osteophytes, spinal stenosis, and endplate changes. Signal intensity was measured quantitatively. RESULTS: After controlling for age, disc height was associated with all back pain variables studied and anular tears with LBP frequency and intensity during the 12 months before imaging. Both were associated with lifetime frequency of low back pain interfering with daily activities, disability, and intensity of the worst lifetime pain episode. Other MRI findings did not explain the various symptom histories. Adjusting for physical loading in the past 12 months increased the associations of anular tears and "low back pain today" and 12-month low back pain parameters. After controlling for genotype and other familial influences, the within-pair differences in disc height and anular tears accounted for 6% to 12% of the total variance in the within-pair differences of low back pain variables. CONCLUSION: These findings raise new questions about the underlying mechanisms of LBP. The sensitivities of the only significant MRI parameters, disc height narrowing and anular tears, are poor, and these findings alone are of limited clinical importance. PMID- 12642767 TI - Familial idiopathic scoliosis: evidence of an X-linked susceptibility locus. AB - STUDY DESIGN: A genomic screen and statistical linkage analysis of a large sample of families with individuals having idiopathic scoliosis was performed. OBJECTIVES: To identify an X-linked susceptibility locus involved in the expression of familial idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: A large sample of families with individuals having idiopathic scoliosis (202 families; 1198 individuals) were diagnosed through physical examination and radiographic criteria, and genomic screening and genetic linkage analyses were performed. METHODS: Model-independent linkage analysis was used to screen genotyping data from 15 X-linked markers in 202 families (1198 individuals). Families were stratified based on the ratio of the likelihood of an X-linked dominant (XLD) inheritance model relative to that of an autosomal dominant (AD) model. Both model-independent and model-dependent linkage analyses were used to identify potential candidate regions. RESULTS: When the entire set of families were analyzed with model-independent methods, no result was significant at the 0.05 level for any of the markers. However, when the families were stratified based on the ratio of the likelihood of the X-linked dominant to autosomal dominant mode of inheritance, results from model-dependent linkage analysis of 15% of the families most likely to have X-linked dominant inheritance showed six adjacent markers with positive lod score values and a maximum lod score of 1.69 (theta = 0.2) at marker GATA172D05. A lod score of 2.23 at this same marker was found in a single family with six affected individuals. CONCLUSION: The results suggest that a region on the X chromosome may be linked to the expression of familial idiopathic scoliosis in a subset of these families. PMID- 12642768 TI - Electrical stimulation for intraoperative evaluation of thoracic pedicle screw placement. AB - STUDY DESIGN: Thoracic pedicle screws were stimulated electrically while recording electromyography (EMG) activity in associated muscle groups intraoperatively. Screw position was then evaluated after surgery using computed tomography (CT). The CT results were compared with evoked EMG threshold values. OBJECTIVE: To report our experience with pedicle screw stimulation in thoracic vertebrae in 22 patients who underwent thoracic level spinal instrumentation from May 1999 to March 2001. SUMMARY OF BACKGROUND DATA: Pedicle screw stimulation for intraoperative assessment of screw placement has been reported in lumbosacral spinal procedures. Evoked EMG thresholds >10 or 11 mA are associated with increased probability of vertebral cortex violation and postoperative complications. Thresholds >15 mA provide a 98% confidence of accurate screw positioning on postoperative CT scans. METHODS: Evoked EMG activity was recorded from ulnar carpal flexors, intercostals, or abdominal muscle groups, depending on the level of screw placement. Postoperative CT scans were read by a staff orthopedic spine surgeon, a senior resident in orthopedics, and a musculoskeletal radiologist. RESULTS: Results of 87 screws are reported. Five screws (5.7%) showed penetration on postoperative CT scans. Six screws (6.9%) had stimulation thresholds < or =11 mA, of which three showed cortical breakthrough. Of the 81 screws with thresholds >11 mA, 79 (97.5%) were within the vertebra. No postoperative neurologic complications were noted in any of the 22 patients. CONCLUSION: These results are consistent with previous studies of lumbosacral pedicle screws. In this series, stimulation thresholds >11 mA have a 97.5% negative predictive value, suggesting that cortical violation is highly unlikely. Although judgment of screw placement should not depend solely on stimulation thresholds, pedicle screw stimulation may provide rapid and useful intraoperative information on screw placement during procedures involving the use of thoracic pedicle screws. PMID- 12642769 TI - Adult scoliosis: a health assessment analysis by SF-36. AB - STUDY DESIGN: A prospective self-assessment analysis of a consecutive series of adult patients diagnosed with adult scoliosis of adolescent onset or de novo degenerative scoliosis. PURPOSE: To analyze the impact that scoliosis has on patient health perceptions. SUMMARY OF BACKGROUND DATA: There exists little data studying the effect scoliosis has on an adult's self-perception of health. METHODS: Inclusion criteria were: age > or =18 years, Cobb angle greater than 10 degrees, degenerative or idiopathic deformity, no prior surgery (spine), and complete records. Each patient completed the standard Short Form-36 (SF-36) questionnaire. For each patient, radiographic and clinical data were evaluated. After all data were collected, the mean, standard deviation (SD), and true N values of the Cobb angle were calculated. This study population was then divided into two groups: patients with a Cobb angle greater than 10 degrees and patients with a Cobb angle greater than 20 degrees. Comparison of the SF-36 data for adult scoliosis patients with the United States general population, the United States general population ages 55-64, and patients with hypertension and low back pain was undertaken. RESULTS: The mean age for the patients in this study was 63 years of age; 22 patients were diagnosed as having adult scoliosis of adolescent onset and 27 patients had de novo degenerative scoliosis. The Cobb angle in this study group was found to be greater than 20 degrees in 41 patients; of these patients, 19 patients had a diagnosis of adult scoliosis of adolescent onset and 22 patients had a diagnosis of de novo degenerative scoliosis. Taken as a whole, the patients in this study averaged scores much lower than the norms for both the general U.S. population in all 8 categories and the U.S. population for ages 55 to 64 in 7 out of 8 categories of the SF-36 questionnaire. Additionally, patients with scoliosis also showed lower scores compared to that of the norms found of patients with comorbid conditions: back pain/sciatica with hypertension in seven of the eight categories: physical functioning, role physical, bodily pain, general health, vitality, social functioning, role emotional, and mental health. Of the radiographic parameters analyzed, loss of lumbar lordosis revealed a significant correlation with social function: P = 0.018; role emotional: P = 0.038; and overall general health: P = 0.05. The other radiographic parameters did not reveal statistically significant correlation to the calculated SF-36 scores. CONCLUSION: Our data clearly demonstrate the impact that adult scoliosis has on a patient's perception of health. The severity of this impact is strikingly apparent in the SF-36 scores when compared to benchmark data on patients with comorbid conditions such as back pain and hypertension. It is our conclusion that adult scoliosis is becoming a medical condition of significant impact, affecting the fastest growing section of our society to a previously unrecognized degree. PMID- 12642770 TI - Is a condition-specific instrument for patients with low back pain/leg symptoms really necessary? The responsiveness of the Oswestry Disability Index, MODEMS, and the SF-36. AB - STUDY DESIGN: Analysis of longitudinal data collected prospectively from patients seen in 27 National Spine Network member centers across the United States. OBJECTIVE: To evaluate the responsiveness of the Oswestry Disability Index, MODEMS scales, and all scales and summary scales of the MOS Short-Form 36 (SF-36) for patients with low back pain/leg symptoms. SUMMARY OF BACKGROUND DATA: The responsiveness of general and condition-specific health status instruments is a key concept for clinicians and scientists. Various authors have explored responsiveness in common surveys used to assess spine patients. Although it is generally believed that condition-specific measures are more responsive to change in the condition under study, in the case of low back pain, most authors agree that further exploration is necessary. METHODS: Patients with diagnoses of herniated disc, spinal stenosis, and spondylosis from the National Spine Network database who completed baseline and 3-month follow-up surveys were analyzed. Patient-provider consensus regarding improvement, worsening, or no change in the condition was selected as the external criterion. Responsiveness was evaluated using ROC curve analysis and effect size calculations. RESULTS: Nine hundred and seventy patients had complete data at baseline and 3 months. At follow-up, 68% of the patients had consensus improvement. Based on ROC analysis, scales assessing pain were significantly more responsive than scales assessing function. There were no significant differences between the condition-specific scales and their equivalent general-health counterpart. The scales with the highest probabilities of correctly identifying patient's improvement were: the condition-specific pain scale from MODEMS (PAIN, ROC = 0.758); the combined pain and function scale from MODEMS (MPDL, ROC = 0.755); the general pain scale from the SF-36 (BP, ROC = 0.753); the combined pain and function scale from the SF-36 (PCS, ROC = 0.745); the condition-specific function measure from the Oswestry (ODI, ROC = 0.723); and the physical function measure from the SF-36 (PF, ROC = 0.721). A similar rank order was typically maintained with effect size calculations. Results were nearly identical in patients with multiple non-spine-related comorbidities and in patients with high degrees of perceived disability. The BP scale was most responsive to worsening of symptoms. CONCLUSION: For studies of patients with low back problems, the general SF-36 may be a sufficient measure of health status and patient function, without the need for additional condition-specific instruments. Pain scales appear to be the most responsive measures in patients with low back pain. PMID- 12642771 TI - Rates of advanced spinal imaging and spine surgery. AB - STUDY DESIGN: Small area analysis. OBJECTIVES: To determine the association between the rates of advanced spinal imaging and spine surgery across geographic areas. SUMMARY OF BACKGROUND DATA: The rates of spine surgery in the United States have increased along with a concurrent rise in the use of advanced spinal imaging: CT and MRI. Spine surgery rates vary six-fold across geographic areas of the United States. Differences in patient populations and health care supply have explained only about 10% of this variation. METHODS: We used a random 5% sample of Medicare's National Claims History Part B files for 1996 and 1997 to determine procedure rates across 306 Hospital Referral Regions. We analyzed the association between spinal imaging and spine surgery using linear regression. Main outcome measures were rates of procedures and coefficients of determination (R2). RESULTS: The rates of advanced spinal imaging (CT and MRI combined) varied 5.5 fold across geographic areas. Areas with higher rates of MRI had higher rates of spine surgery overall (r = 0.46) and spinal stenosis surgery specifically (r = 0.37). The rates of advanced spinal imaging accounted for 22% of the variability in overall spine surgery rates (R2 = 0.22, P < 0.001) and 14% of the variability in lumbar stenosis surgery rates (R2 = 0.14, P < 0.001). A simulation model showed that MRIs obtained in the patients undergoing surgery accounted for only a small part of the correlation between MRI and total spine surgery rates. CONCLUSIONS: A significant proportion of the variation in rates of spine surgery can be explained by differences in the rates of advanced spinal imaging. The indications for advanced spinal imaging are not firmly agreed on, and the appropriateness of many of these imaging studies has been questioned. Improved consensus on the use and interpretation of advanced spinal imaging studies could have an important effect on variation in spine surgery rates. PMID- 12642772 TI - Risk of multiple reoperations after lumbar discectomy: a population-based study. AB - STUDY DESIGN: Retrospective follow-up study of patients undergoing multiple (two or more) reoperations after initial lumbar discectomy using an administrative database. OBJECTIVES: To identify the population-based risk of multiple reoperations after lumbar discectomy and to analyze factors associated with the risk. SUMMARY OF BACKGROUND DATA: Although multiple reoperations after initial lumbar discectomy are likely uncommon, research to better understand reasons for and outcomes of reoperations is needed because of the large number of discectomies performed. METHODS: Data on all lumbar spine operations during 1987 1998 were obtained from the Finnish Hospital Discharge Register. The patient's initial disc operation during the study period was linked to subsequent operations, and patients with two or more reoperations were analyzed further. The risk of multiple reoperations was determined using the methods of event history analysis. RESULTS: Among 35,309 patients undergoing an initial discectomy, 4943 (14.0%) had at least one reoperation and 803 (2.3%) had two or more reoperations. A total of 63% of the second reoperations were discectomies, 14% were fusions, and the remaining 23% were decompressions. Patients with one reoperation after lumbar discectomy had a 25.1% cumulative risk of further spinal surgery in a 10 year follow-up. Reduced risk was seen when the first reoperation took place more than 1 year after the initial discectomy (relative risk 0.83, 95% confidence interval 0.72-0.96), in patients for whom the first reoperation had been a fusion (relative risk 0.27, 95% confidence interval 0.12-0.61), and in patients 50-64 years of age (relative risk 0.62, 95% confidence interval 0.48-0.79). CONCLUSION: Patients with one reoperation after lumbar discectomy are at considerable risk of further spinal surgery. PMID- 12642773 TI - Psoas abscess: the spine as a primary source of infection. AB - STUDY DESIGN: Case report, literature review, discussion. OBJECTIVES: To emphasize the role of the spine as primary source of infection for psoas abscess. SUMMARY OF BACKGROUND DATA: Spine-associated psoas abscesses increase with more frequent invasive procedures of the spine and recurring tuberculosis in industrialized countries. Diagnosis is often delayed by misinterpretation as arthritis, joint infection, or urologic or abdominal disorders. METHODS: We present six cases of psoas abscesses associated with spinal infections that were treated in our hospital from January to December 2001. Diagnostic and treatment concepts are discussed. RESULTS: Our data emphasize the importance of the spine as primary source of infection and suggest an increase in the incidence of secondary psoas abscess. Treatment includes open surgical drainage and antibiotic therapy. In patients with high operative risk and uniloculated abscess, a CT guided percutaneous abscess drainage can be sufficient. It is essential to combine abscess drainage with causative treatment of the primary infectious focus. Related to the spine, this includes treatment of spondylodiscitis or implant infection after spinal surgery. Usually, several operations are necessary to eradicate bone and soft-tissue infection and restore spinal stability. Continuous antibiotic therapy over a period of 2-3 weeks after normalization of infectious parameters is recommended. CONCLUSION: The spine as primary source of infection for secondary psoas abscess should always be included in differential diagnosis. Because the prognosis of psoas abscess can be improved by early diagnosis and prompt onset of therapy, it needs to be considered in patients with infection and back or hip pain or history of spinal surgery. PMID- 12642774 TI - Unusual metastasis to the cauda equina from renal cell carcinoma. AB - STUDY DESIGN: This is a case report of a patient with renal cell carcinoma and intradural metastasis to the cauda equina. OBJECTIVE: To present a rare case of an intradural metastasis from renal cell carcinoma and to discuss the clinical features of metastatic tumors in the cauda equina and the possible mechanism of the tumor spread to the cauda equina. SUMMARY OF BACKGROUND DATA: Intradural spinal metastasis has been rarely reported in the English literature. Only two reports that describe the spread of metastatic renal cell carcinoma to the cauda equina have been published. METHODS: A 61-year-old man who underwent a nephrectomy for the treatment of renal cell carcinoma presented with worsening low back pain that radiated to both legs. Magnetic resonance imaging showed an ill-defined tumor mass in the cauda equina at L3. After a recapping T-saw laminoplasty of L3 was performed, the tumor was excised and the nerves involved in the tumor were transected. RESULTS: Pathologic examination showed papillary renal cell carcinoma with identical histology to the primary tumor. The patient's low back pain and radiating leg pain were relieved after operating. CONCLUSION: The majority of cauda equina tumors are primary tumors, and metastases are very rare. Intradural spinal metastasis pain is a characteristic cramping pain provoked by light percussion on the lumbar spine, becoming severe when sleeping in the flexion or sitting position. Magnetic resonance imaging is a useful tool for detecting intraspinal metastasis when the patient is complaining of a unique pain. A recapping T-saw laminoplasty to preserve posterior elements with tumor removal is feasible for relieving pain and demonstrating the pathology. PMID- 12642776 TI - Massive intestinal hemorrhage resulting from a polypoid tumor in the sigmoid colon: an unusual complication of a giant cell tumor of the fifth lumbar vertebra. AB - STUDY DESIGN: Clinicopathologic study of a case of giant cell tumor of the spine. SUMMARY OF BACKGROUND DATA: Giant cell tumors of the spine are uncommon, accounting for 1.3-6.5% of all cases in various series. Because of their location, they may cause neurologic deficits. The treatment consists of excision or curettage and has been claimed to give good results. METHODS: A 33-year-old woman presented with low back pain in 1995; radiologic investigation and biopsy showed features of giant cell tumor involving the fifth lumbar and first sacral vertebrae. Wide excision was performed, but the tumor recurred in 1996 and was curetted. She developed massive intestinal bleeding in 1997 resulting from infiltration of the sigmoid colon by giant cell tumor in the form of a polypoid intraluminal mass. The involved segment of colon was resected, and the patient remained alive, although debilitated, 7 years after initial presentation. RESULTS: Examination of the tumor in the spine and the colon showed typical histology of giant cell tumor with no evidence of malignant transformation. The involved colon was freely mobile and away from the tumor of the spine. CONCLUSION: Giant cell tumor of the spine can result in unusual complication, massive intestinal hemorrhage in our case, which causes considerable morbidity. PMID- 12642775 TI - Medial compartment syndrome of the foot: an unusual complication of spine surgery. AB - STUDY DESIGN: Descriptive case report. OBJECTIVES: To report the case of a child with medial compartment syndrome of the foot following posterior spinal instrumentation and fusion. SUMMARY OF BACKGROUND DATA: No previous study has reported medial compartment syndrome of the foot following spinal surgery. METHODS: A 15-year-old female with progressive idiopathic scoliosis was taken for posterior instrumentation and fusion. The patient had a history of severe postexertional cramping in the feet following athletics. Surgery progressed uneventfully and the patient was continuously monitored with somatosensory-evoked potentials, which showed no changes. In the recovery room, the patient complained of severe cramping in one foot that was similar to her postexertional cramping. This was lessened with massage and ketorolac. Soreness continued in the foot into postoperative day one and then increased overnight. On the morning of postoperative day 2, pressure in the medial compartment was found to be 97 mm Hg and she was taken for fasciotomy, which found necrosis of the abductor hallucis muscle, and all other compartments of the foot were normal. RESULTS: At the 6 month follow-up, the patient is doing well with no known sequelae. CONCLUSION: This was a very rare case of medial compartment syndrome of the foot following spine surgery. We believe that the patient had a predisposition, whether neurologic or vascular, toward cramping in the foot and that this activity was stimulated by the nerve stimulation during the evoked potential monitoring. Although the patient had thoracic epidural analgesia after surgery, it was not felt to have contributed to the development or result of the compartment syndrome. We strongly advocate for checking patients feet and legs during surgery for overactivity and stress the need for a high index of suspicion for compartment syndrome for unexplained pain after surgery. PMID- 12642777 TI - Nontraumatic acute complete paraplegia resulting from cervical disc herniation: a case report. AB - STUDY DESIGN: A case report of nontraumatic acute complete paraplegia resulting from cervical disc herniation. OBJECTIVES: To describe a rare case of nontraumatic paraplegia resulting from enlargement of a herniated disc in the cervical spine and to outline appropriate management of a patient with severe spinal cord compression secondary to disc herniation with developmental spinal canal stenosis. SUMMARY OF BACKGROUND DATA: Acute progression of myelopathy into complete paraplegia resulting from disc herniation is rare. There are only four reported cases of nontraumatic acute myelopathy secondary to cervical disc herniation. No other report has described magnetic resonance imaging findings noted before and after the onset of acute myelopathy. METHODS: A cervical disc herniation at C6-C7 is reported in a 29-year-old man who had nontraumatic acute complete paraplegia. Neurologic and magnetic resonance imaging findings are evaluated and discussed. RESULTS: Disc herniation at C6-C7 enlarged nontraumatically, resulting in complete paraplegia. Emergent anterior decompression followed by secondary posterior multilevel decompression was performed. Magnetic resonance imaging studies revealed localized high signal intensity change in the spinal cord. No neurologic recovery was achieved 3 years post-surgery. CONCLUSION: We emphasize that there is a possibility of acute, irreversible progression of paralysis secondary to nontraumatic enlargement of cervical disc herniation with canal stenosis. In these cases, immediate early decompressive surgery is crucial to the prevention of severe myelopathy. PMID- 12642778 TI - 'Omic' approaches and lipid metabolism: are these new technologies holding their promises? PMID- 12642779 TI - Molecular mechanisms of autosomal recessive hypercholesterolemia. AB - PURPOSE OF REVIEW: Autosomal recessive hypercholesterolemia (ARH) is a rare Mendelian dyslipidemia characterized by markedly elevated plasma LDL levels, xanthomatosis, and premature coronary artery disease. LDL receptor function is normal, or only moderately impaired in fibroblasts from ARH patients, but their cultured lymphocytes show increased cell-surface LDL binding, and impaired LDL degradation, consistent with a defect in LDL receptor internalization. Recently, the disorder was shown to be caused by mutations in a phosphotyrosine binding domain protein, ARH, which is required for internalization of low density lipoproteins in the liver. This review summarizes the findings of new investigations into the pathophysiology and molecular genetics of ARH. RECENT FINDINGS: All mutations that have been characterized to date preclude the synthesis of a full-length protein. GST-pulldown experiments indicate that the phosphotyrosine binding domain of ARH interacts with the internalization sequence (NPVY) in the cytoplasmic tail of LDLR, and that conserved motifs in the C terminal portion of the protein bind to clathrin and to the beta2-adaptin subunit of AP-2. SUMMARY: The available data suggest that ARH functions as an adaptor protein that couples LDLR to the endocytic machinery. PMID- 12642780 TI - Peroxisome proliferator-activated receptors beta/delta: emerging roles for a previously neglected third family member. AB - PURPOSE OF REVIEW: Peroxisome proliferator-activated receptors alpha, beta/delta and gamma are members of the nuclear receptor superfamily. They mediate the effects of fatty acids and their derivatives at the transcriptional level, and are considered to be lipid sensors that participate in the regulation of energy homeostasis. Compared with the alpha and gamma peroxisome proliferator-activated receptor isotypes, peroxisome proliferator-activated receptor beta functions have long remained an enigma. In this review, we focus on emerging knowledge about peroxisome proliferator-activated receptor beta activation and roles. RECENT FINDINGS: We review recent data that suggest key roles in basic cell functions, such as proliferation, differentiation and survival, and in embryonic development and lipid metabolism in peripheral tissues. SUMMARY: The newly unveiled roles of peroxisome proliferator-activated receptor beta in important basic cell functions certainly justify a further exploration of its potential as a therapeutic target in pathologies such as metabolic syndrome X or skin diseases. PMID- 12642781 TI - Liver X receptors: new players in atherogenesis? AB - PURPOSE OF REVIEW: The liver X receptors (alpha and beta) are oxysterol-activated nuclear receptors. A large number of liver X receptor target genes with functions in lipid homeostasis have been identified. Increasing evidence indicates that these receptors play a beneficial role in avoiding cholesterol overload by intervening at several steps of cholesterol metabolism. In this review, we describe the most recent developments concerning their functions in cholesterol and lipid metabolism and their impact in atherogenesis. RECENT FINDINGS: Although potentially undesirable effects of liver X receptor activation on triglyceride metabolism have been observed, recent in-vivo studies confirm that the overall trend is a reduction of atherogenesis. SUMMARY: Consequently, liver X receptors are becoming therapeutic targets of great interest for the treatment of atherosclerosis, especially if their action on triglyceride and cholesterol metabolism can be dissociated and isoform-specific ligands identified. PMID- 12642782 TI - Role of estrogen receptor-alpha in pharmacogenetics of estrogen action. AB - PURPOSE OF REVIEW: To summarize recent data regarding the role of estrogen receptor-alpha polymorphisms in determining the response to estrogen therapy or the risk of clinical cardiovascular events. RECENT FINDINGS: Recent clinical trials of hormone replacement therapy for cardiovascular disease have yielded surprisingly negative results, shifting clinical opinions from a position of presumed cardiovascular benefit to one of confirmed harm. Understanding why hormone replacement therapy has beneficial effects on intermediate risk markers for cardiovascular disease, but produces an increase in cardiovascular events, is an important public health question with the potential to elucidate fundamentally important aspects on atherogenesis, cardiovascular disease, and the biology of estrogen action. One question concerning the cardiovascular effects of hormone replacement therapy is whether genetic factors can substantially modify individual responses to estrogen treatment. New clinical trial evidence is emerging that links the presence of particular variants in the estrogen receptor to the response of HDL and other intermediate endpoints to hormone replacement therapy. SUMMARY: One or more common variants in estrogen receptor-alpha are associated with a differential response to hormone replacement therapy in several domains of estrogen action. However, the effect of these variants on the risk of clinical cardiovascular events in the setting of hormone replacement therapy is not yet known. Additional research focusing on the clinical impact of common variants in estrogen receptor-alpha, estrogen receptor-beta and the progesterone receptor promise to improve clinical decision-making concerning the use of hormone replacement therapy and other novel estrogen agonists. PMID- 12642783 TI - Structure-function relationships of apolipoprotein A-I: a flexible protein with dynamic lipid associations. AB - PURPOSE OF REVIEW: Apolipoprotein A-I is the major structural protein of HDL. Its physicochemical properties maintain a delicate balance between maintenance of stable lipoproteins and the ability to associate with and dissociate from the lipid transported. Here we review the progress made in the last 2-3 years on the structure-function relationships of apolipoprotein A-I, including elements related to the ATP binding cassette transporter A1. RECENT FINDINGS: Current evidence now supports the so-called 'belt' or 'hairpin' models for apolipoprotein A-I conformation when bound to discoidal lipoproteins. In-vivo expression of apolipoprotein A-I mutant proteins has shown that both the N- and C-terminal domains are important for lipid association as well as for the esterification reaction, particularly binding of cholesteryl esters and formation of mature alpha-migrating lipoproteins. This property is apparently quite distinct from the activation of the enzyme lecithin cholesterol acyl transferase, which requires interaction with the central helix 6. The interaction of apolipoprotein A-I with the ATP binding cassette transporter A1 has been shown to require the C-terminal domain, which is proposed to mediate the opening of the helix bundle formed by lipid-free or lipid-poor apolipoprotein A-I and allow its association with hydrophobic binding sites. SUMMARY: Significant progress has been made in the understanding of the molecular mechanisms controlling the folding of apolipoprotein A-I and its interaction with lipids and various other protein factors involved in HDL metabolism. PMID- 12642784 TI - Apolipoprotein A-I(Milano): current perspectives. AB - PURPOSE OF REVIEW: Strategies to increase HDL are among the major targets of clinical research in atherosclerosis prevention. The mutant apolipoprotein A I(Milano) has been associated with a reduced incidence of coronary disease in carriers. Furthermore, recombinant apolipoprotein A-I(Milano) has displayed remarkable atheroprotective activities and the possibility of directly reducing the burden of atherosclerosis in experimental models. This review is aimed at providing an update on the experimental studies in which apolipoprotein A I(Milano), produced as a recombinant protein, has displayed important effects in the treatment of vascular diseases. RECENT FINDINGS: In the past year, two reports have appeared, indicating that a single-dose administration of recombinant apolipoprotein A-I(Milano) dimers formulated into liposomes can reduce atheromas in models such as the apolipoprotein E-deficient mice and a rabbit model of carotid focal lesion, in which a direct 90 min infusion of the product reduced atheroma up to 30%. This finding was associated with an increase in HDL free cholesterol and the permanence of the recombinant product in the lesion for over 72 h. SUMMARY: Recombinant apolipoprotein A-I(Milano), formulated as synthetic HDL with phospholipids, appears to exert a direct removing effect on arterial cholesterol. This is well evident in experimental animals and, more recently in clinical findings, as indicated by a dramatic increase in HDL free cholesterol after the infusion of different doses of the agent. As the product appears to be well tolerated and non-immunogenic, ongoing phase II studies in patients are being awaited with interest to obtain a 'proof of principle' for 'HDL therapy'. PMID- 12642785 TI - Apolipoprotein A-II: beyond genetic associations with lipid disorders and insulin resistance. AB - PURPOSE OF REVIEW: Apolipoprotein A-II, the second major HDL apolipoprotein, was often considered of minor importance relatively to apolipoprotein A-I and its role was controversial. This picture is now rapidly changing, due to novel polymorphisms and mutations, to the outcome of clinical trials, and to studies with transgenic mice. RECENT FINDINGS: The -265 T/C polymorphism supports a role for apolipoprotein A-II in postprandial very-low-density lipoprotein metabolism. Fibrates, which increase apolipoprotein A-II synthesis, significantly decrease the incidence of major coronary artery disease events, particularly in subjects with low HDL cholesterol, high plasma triglyceride, and high body weight. The comparison of transgenic mice overexpressing human or murine apolipoprotein A-II has highlighted major structural differences between the two proteins; they have opposite effects on HDL size, apolipoprotein A-I content, plasma concentration, and protection from oxidation. Human apolipoprotein A-II is more hydrophobic, displaces apolipoprotein A-I from HDL, accelerates apolipoprotein A-I catabolism, and its plasma concentration is decreased by fasting. Apolipoprotein A-II stimulates ATP binding cassette transporter 1-mediated cholesterol efflux. Human and murine apolipoprotein A-II differently affect glucose metabolism and insulin resistance. A novel beneficial role for apolipoprotein A-II in the pathogenesis of hepatitis C virus has been shown. SUMMARY: The hydrophobicity of human apolipoprotein A-II is a key regulatory factor of HDL metabolism. Due to the lower plasma apolipoprotein A-II concentration during fasting, measurements of apolipoprotein A-II in fed subjects are more relevant. More clinical studies are necessary to clarify the role of apolipoprotein A-II in well-characterized subsets of patients and in the insulin resistance syndrome. PMID- 12642786 TI - Cholesteryl ester transfer protein: gathering momentum as a genetic marker and as drug target. AB - PURPOSE OF REVIEW: Cholesteryl ester transfer protein facilitates the exchange of neutral lipids between HDL and apolipoprotein B containing lipoproteins, which hold powerful opposing roles as risk factors for coronary artery disease. The question as to whether cholesteryl ester transfer protein promotes or protects from atherosclerosis, however, has not been answered. RECENT FINDINGS: This review considers studies dealing with cholesteryl ester transfer protein variants and their effect on blood lipids in various metabolic and clinical settings. Other studies discussed deal with the association between the transfer protein and cardiovascular disease. Research on the biological activity of the cholesteryl ester transfer protein molecule is described including a first clinical study where pharmacological inhibition of the protein proved to be effective in raising HDL cholesterol. SUMMARY: Data concerning the potential marker role of cholesteryl ester transfer protein, although accumulating, are still inconclusive and, at present, not useful for clinical decision making. Inhibition of the protein was demonstrated to be feasible and appears to be promising. PMID- 12642787 TI - Hepatic lipase: a marker for cardiovascular disease risk and response to therapy. AB - PURPOSE OF REVIEW: Hepatic lipase plays a key role in the metabolism of pro atherogenic and anti-atherogenic lipoproteins affecting their plasma level as well as their physico-chemical properties. However, controversial evidence exists concerning whether hepatic lipase is pro or anti-atherogenic. The goal of this review is to summarize recent evidence that connects the enzyme to cardiovascular disease. The potential impact of genetic determinants of hepatic lipase activity in modulating both the development of coronary and carotid atherosclerosis will be discussed based on hepatic lipase proposed roles in lipoprotein metabolism. RECENT FINDINGS: Twenty to 30% of individual variation of hepatic lipase activity is accounted for by the presence of a common polymorphism in the promoter region (-514 C to T) of the hepatic lipase gene (LIPC). This polymorphism, via its impact on hepatic lipase synthesis and activity, appears to contribute to (1) individual susceptibility to cardiovascular disease: the presence of the T allele (low hepatic lipase activity) may carry a marginally increased risk of atherosclerosis; (2) carotid plaque composition and individual susceptibility to cerebrovascular events: the presence of the C allele (high hepatic lipase activity) is associated with increased carotid intima-media thickness and abundance of macrophages in the carotid plaque (unstable plaque); and (3) response of cardiovascular disease patients to lipid-lowering therapy: patients with the CC genotype have the greatest clinical benefit from intensive lipid lowering therapy. SUMMARY: Convincing evidence shows that hepatic lipase plays a key role in remnant lipoprotein catabolism as well as in remodeling of LDL and HDL particles. The anti or pro-atherogenic role of hepatic lipase is likely to be modulated by the concurrent presence of other lipid abnormalities (i.e. increased LDL cholesterol levels) as well as by the genetic regulation of other enzymes involved in lipoprotein metabolism. Characterization of patients by their LIPC genotype will contribute to a better definition of individual risk of coronary and cerebrovascular events, specifically in patients with qualitative (small, atherogenic LDL and low HDL2 cholesterol) rather than quantitative lipid abnormalities for whom the routine lipid profile may underestimate the risk of coronary and cerebrovascular disease. PMID- 12642788 TI - Genetics of intima-media thickness. AB - PURPOSE OF REVIEW: Increased carotid artery intima-media thickness is associated with an increased risk of coronary heart disease or cerebrovascular disease and is a powerful predictor of cardiovascular and cerebrovascular outcomes. Consequently, the measurement is now used in a number of case control, cohort and familial and linkage studies as an intermediate phenotype for the investigation of the genetic and environmental determinants of atherosclerosis. The aim of this paper is to review the most recent available data on the genetic determinants of carotid intima-media thickness. RECENT FINDINGS: Genes could account for a significant amount of variation in carotid intima-media thickness: up to 30-66%. Carotid intima-media thickness progressed more rapidly with age in familial hypercholesterolemia patients than in patients without his condition. Familial hypercholesterolemia patients with a null LDL receptor allele tended to have higher carotid intima-media thickness than patients carrying the LDL receptor defective allele. Small association studies showing positive or negative results with the angiotensinogen gene variants as well as with the angiotensin converting enzyme I/D polymorphism add to the confusion in this largely controversial area. Differing results may depend on the vascular territory and genetic background. New associations have been described in small studies. SUMMARY: Studies on the association of polymorphisms and intima-media thickness are frequently disappointing and lead occasionally to conflicting results. Among study limitations is the fact that mostly single gene effects are considered; longitudinal cohort studies may be more appropriate than case control studies. Ongoing large prospective population studies and clinical trials have integrated the measurements of intima-media thickness and genotype determination with a genomic approach. As a result, in the near future we may see more important and robust results with significant consequences on our understanding of genetic determinants of atherosclerotic cardiovascular disease. PMID- 12642789 TI - Nutrition and metabolism. PMID- 12642790 TI - Genetics and molecular biology. PMID- 12642791 TI - Hyperlipidemia and cardiovascular disease. PMID- 12642792 TI - Therapy and clinical trials. PMID- 12642794 TI - Test your knowledge: antineoplastic therapy. PMID- 12642795 TI - Understanding the immune system: overview for infusion assessment. AB - More than 90% of patients who are admitted to an acute care setting or home and ambulatory care services receive some form of infusion therapy that is integral to their treatment plan. The patient's immune system status is an important variable to consider in the assessment, planning, intervention, and evaluation for the multiple and varied types of infusion therapies the patient may require. Infusion nurses have a major role in the provision of these infusion therapies and a responsibility to be knowledgeable about the immune system. PMID- 12642796 TI - Development of a Vascular Access Team in an acute care setting. AB - In today's ever-changing acute care setting, justifying the allocation of resources for development and implementation of a Vascular Access Team (VAT) can often be challenging. The Infusion Nurses Society (2000), which establishes standards of practice for the infusion nursing profession, states that "the profession itself must seek control of its practice to assure the quality of its services to the public. A profession's concerns for quality services reflects its commitment to serving the community." This article discusses the steps that were taken in the development of a VAT in the acute care setting of Froedtert Memorial Lutheran Hospital, a teaching hospital affiliated with the Medical College of Wisconsin. PMID- 12642798 TI - Complications associated with implantable vascular access devices in the patient with cancer. AB - Implantable vascular access devices (IVADs), or implantable ports, have been used for patients with cancer for more than 20 years. Although these devices have greatly improved infusion access for this population of patients, complications may still occur. Nurses working with oncology patients who have IVADs must be aware of possible problems for these patients, as well as the importance of early assessment and intervention. The diagnosis of cancer and potential treatments for this disease may make these patients at higher risk for complications associated with the use of IVADS. PMID- 12642797 TI - The effect of a mediastinal mass on the initial positioning of a peripherally inserted central venous catheter. AB - The purpose of this project was to assess if the presence of a mediastinal mass adversely influences peripherally inserted central catheter (PICC) positioning. A retrospective review of all PICC placements over an 18-month period in a pediatric oncology hospital was conducted in which patients were categorized by the presence or absence of a mediastinal mass. A much higher proportion of patients with a mediastinal mass (8 of 13) had a malpositioned catheter than those without a mass (3 of 38; P = .0002). A significant proportion of the malpositioned catheters in patients with a mediastinal mass terminated in the right heart chambers. These findings show that clinicians must be vigilant about checking for PICC malpositioning in this patient population. PMID- 12642799 TI - Pharmacological versus nonpharmacological techniques in reducing venipuncture psychological trauma in pediatric patients. AB - Peripherally inserted central catheters (PICCs) provide reliable access for pediatric patients requiring infusion therapy. Our multidisciplinary team includes nurses, physicians, child life specialists, and radiology technicians. In preparing and educating the patient and parent for the procedure, cultural and developmental levels are assessed and planned accordingly. PMID- 12642800 TI - 10 lessons on writing for publication. AB - All nurses have the potential to be an author. Each nurse can make wonderful contributions to the body of nursing literature, whether you're a staff nurse, educator, manager, or even a student. Your perspective can help others gain insights into our profession. The biggest barrier to writing for a nursing journal is that nurses assume that authors easily come up with great ideas and write wonderful articles. That's not how it works! Investigation and preparation before writing play a large part in any successful author's work. This work is time-consuming, I admit, but if you do it, you'll be better prepared to write and you'll increase your chances of having your manuscript accepted by a journal. The information in this article will help you write for both formal and informal style journals. PMID- 12642801 TI - The challenge of articulating what we do. PMID- 12642802 TI - Defining our practice: a personal perspective. PMID- 12642803 TI - Using technology to advance the knowledge work of clinical nurse specialists: part I: Organizing data. PMID- 12642804 TI - Clotting and bleeding: anticoagulant therapy update for the clinical nurse specialist. PMID- 12642805 TI - Entrepreneurship: start-up questions. PMID- 12642806 TI - Sexual relationships, condom use, and concerns about pregnancy, HIV/AIDS, and other sexually transmitted diseases. AB - As we move further into the 21st century, there are increasing numbers of teenagers and young adults infected with sexually transmitted diseases and acquired immunodeficiency syndrome. Aside from sexual abstinence, condom use is the best way to protect oneself from sexually transmitted diseases, including human immunodeficiency virus/acquired immunodeficiency syndrome. Participants in this study were predominantly female, predominantly Caucasian psychology students who experienced sexual intercourse with an opposite sex partner in the past year. Slightly fewer than half of these participants reported condom use at their last episode of vaginal intercourse and type of partner did not affect their condom use, nor did concern about human immunodeficiency virus/acquired immunodeficiency syndrome and sexually transmitted diseases. Findings such as these challenge advanced practice nurses to generate innovative strategies to promote condom use in all types of relationships. We propose that teaching about the link between cervical cancer and lack of condom use could be one of these new strategies. PMID- 12642808 TI - Primary prevention programs to reduce heart disease risk in women. AB - Cardiovascular disease is the number one killer of women. Despite this statistic, many women do not perceive heart disease as a priority health concern. Recognizing that prevention begins in the community, the American Hospital Association launched a national fellowship program in 1998 to improve cardiovascular health. This article describes a practice innovation designed by a clinical nurse specialist who participated as a 1999-2000 cardiovascular fellow. The Women's Prevention Center, a community-based clinic within the walls of a heart hospital, provides a personalized cardiovascular assessment for women, including both heart history and assessment of physical parameters (eg, blood pressure, fasting glucose, lipid panel, waist-hip ratio). From this assessment, women are counseled on strategies to modify their individual risk factors, such as smoking cessation, heart-healthy nutrition and exercise, and are assisted in developing realistic lifestyle or medical management goals. After the initial assessment, outcomes are measured at 3-, 6-, and 12-month follow-ups to determine both individual success with behavior changes and program effectiveness. PMID- 12642807 TI - Psychiatric-mental health clinical nurse specialist practice in a public school setting. AB - The public school setting is increasingly becoming an entry point of contact for an array of healthcare services because it is most likely the best positioned institution to improve a community's overall healthcare. By linking behavioral healthcare services with primary healthcare services in the school setting, the psychiatric-mental health clinical nurse specialist is well suited to provide comprehensive behavioral health services. PMID- 12642809 TI - Alhabeeb's calligraphy "Peace and beauty": a bridge to understanding. PMID- 12642818 TI - Noninjection routes for immunotherapy. AB - Allergen specific immunotherapy, together with drugs and allergen avoidance, is a cornerstone in the management of respiratory allergy. The traditional subcutaneous route is burdened with the risk of severe adverse events; therefore, safer routes of administration (noninjection or local routes) have been investigated and developed. Controlled trials failed to demonstrate the clinical efficacy and the safety of oral and bronchial administration, and these routes have been abandoned. Local nasal immunotherapy proved effective and safe in 17 of 18 controlled trials; thus it is considered a viable route of immunotherapy. Nevertheless, nasal immunotherapy is effective in rhinitis only and requires a particular administration technique; therefore its use is slowly declining. The sublingual route is supported by numerous controlled trials showing its efficacy in asthma and rhinitis in adults and children. The safety profile, assessed in clinical trials and postmarketing surveillance studies, is satisfactory; the most frequent side effects are gastrointestinal complaints, which can be easily managed by proper dose adjusting. Sublingual immunotherapy is now accepted by the World Health Organization as a valid alternative to the subcutaneous route also in children. Although the long-lasting efficacy has been recently documented for the sublingual route, several points still need to be elucidated, including mechanisms of action, optimal dosage, cost-effectiveness, and adherence. PMID- 12642820 TI - The role of T lymphocytes in the pathogenesis of asthma. AB - There is considerable evidence to support a role for T cells in asthma, particularly the involvement of T(H)2 cells both in atopic allergic asthma and in nonatopic and occupational asthma. There might also be a minor contribution from T(C)2 CD8+ T cells. Several T(H)2 cytokines have the potential to modulate airway inflammation, particularly IL-13, which induces airway hyperresponsiveness independently of IgE and eosinophilia in animal models. The identification of transcription factors controlling T(H)1 and T(H)2 development further support the T(H)2 hypothesis because GATA3 is overexpressed and T-bet is underexpressed in the asthmatic airway. Specific T cell directed immunotherapy might allow induction, modulation, or both of T-cell responses, and elucidation of the mechanisms of regulatory T cells might allow further optimization of immunotherapy. Recent advances in our understanding of dendritic cell function in directing T-cell responses might uncover further therapeutic targets. The efficacy of cyclosporin A and anti-CD4 treatment in patients with chronic severe asthma argues for continued T-cell involvement, but whether remodeling contributes to pathology inaccessible to anti-inflammatory treatment or T-cell immunotherapy will be an important future question. PMID- 12642822 TI - Genetic association studies: what are they teaching clinicians? PMID- 12642823 TI - Dietary prevention of allergy, atopy, and allergic diseases. PMID- 12642824 TI - Hygiene hypothesis: fact or fiction? AB - The hygiene hypothesis of asthma and allergy has recently received a swell of popularity and published supporting evidence, and has been extended to autoimmune conditions of childhood. Broadly stated, naturally occurring infections and microbial exposures might essentially immunize against the development of asthma and allergic and autoimmune diseases. If true, then reductions in nature's immunotherapy over the past century might be a major factor in the global increase of these conditions (eg, the higher prevalence of asthma and allergies in urban metropolitan areas compared with rural and farm communities) and might lead to new therapies for these conditions. Although such a unifying hypothesis has great appeal, currently it is only speculation about what might be at the end of the investigative road. How close are the current studies to establishing a causal relationship between microbial exposures and a reduction in allergic, asthmatic, and autoimmune disease prevalence? A systematic epidemiologic appraisal of the current hygiene hypothesis evidence can provide a critical analysis of what is currently known and an investigative blueprint for future studies that can ultimately prove causation and improve recommendations, interventions, and therapies. PMID- 12642826 TI - A questionnaire to measure quality of life in adults with nocturnal allergic rhinoconjunctivitis. AB - BACKGROUND: Many patients with rhinoconjunctivitis experience symptoms at night and these interfere with getting a good night's sleep. OBJECTIVE: The aim of this study was to develop the Nocturnal Rhinoconjunctivitis Quality of Life Questionnaire (NRQLQ) to measure the functional problems that are most troublesome to patients with nocturnal allergic rhinoconjunctivitis (NAR). METHODS: After generating a list of 256 potentially important items, 132 patients with NAR identified the items that were most troublesome. The top scoring 16 items are included in the NRQLQ in four domains (sleep problems, symptoms during sleep time, symptoms on waking and practical problems). The NRQLQ asks patients to recall their experiences during the previous week and score each item on a 7 point scale. RESULTS: One hundred and six patients with NAR participated in a 3 week validation study. This showed the NRQLQ to have good reliability (ICC = 0.87) and responsiveness (responsiveness index = 0.93). For both cross-sectional and longitudinal construct validity correlations between the NRQLQ and other measures of health status were close to a priori predictions. CONCLUSIONS: The NRQLQ has strong evaluative and discriminative measurement properties and can be used with confidence in both clinical practice and clinical trials to measure the functional impairments that are most troublesome to patients with nocturnal allergic rhinoconjunctivitis. PMID- 12642825 TI - Fetal safety of loratadine use in the first trimester of pregnancy: a multicenter study. AB - BACKGROUND: Women in their childbearing years often require drug therapy for allergic conditions. Loratadine, a newer nonsedating antihistamine, is often used because of its preferred side effect profile. To date no published data exist on the safety of loratadine use in pregnancy. OBJECTIVE: We sought to determine whether the use of loratadine in the first trimester of pregnancy was associated with an increased risk for major malformations. Secondary outcomes included rates of miscarriage, birth weights, and gestational age at delivery. METHODS: All women were prospectively enrolled from 4 participating centers. Detailed maternal medical history and drug exposures were collected at intake, whereas pregnancy complications and outcomes were collected at follow-up. A group of unexposed control subjects were recruited and followed up in a similar manner. RESULTS: This report includes follow-up on 161 loratadine exposed pregnancies and an equal number of unexposed control subjects. Maternal characteristics (age, pregnancy history, alcohol consumption, and smoking habits) were not different between the 2 groups. There were 5 malformations observed in the exposed group and 6 in the control group, which was not significantly different (P =.9) Similarly, the live birth rate, gestational age at delivery, and birth weights were not different between the 2 groups. CONCLUSION: These results suggest that loratadine use in pregnancy is not associated with a large risk for major malformations. Further studies are warranted to confirm these findings and to increase study power. PMID- 12642827 TI - Development of a quality-of-life assessment for the allergic child or teenager with multisystem allergic disease. AB - BACKGROUND: Health-related quality of life (HRQOL) questionnaires currently being used to evaluate allergic disease are organ-specific. They therefore fail to take account of the systemic aspects of allergic disease. OBJECTIVE: To develop and validate a pediatric HRQOL questionnaire for allergic disease (Pediatric Allergic Disease Quality of Life Questionnaire, PADQLQ) that encapsulates problems related to the eyes, ears, nose, lungs, skin, emotions, and everyday activities. METHODS: In the development phase, 77 subjects (6 to 16 years of age), with seasonal or perennial allergic problems, were asked how much they were bothered by each different area of HRQOL impairment. The highest scoring areas were used to construct the PADQLQ. In the validation phase of the study, 36 subjects (8 to 16 years of age) with seasonal allergic rhinoconjunctivitis, seasonal allergic asthma, and/or cutaneous manifestations of grass pollen allergy were assessed before and during the pollen season. RESULTS: The PADQLQ contains 26 questions. In addition to standard symptoms (eg, rhinitis), it incorporates multiorgan symptoms that are usually overlooked (eg, hearing problems). The PADQLQ demonstrated good cross-sectional and longitudinal validity, showing a high degree of correlation with symptom scores and quality of life as measured by a visual analogue scale and two-organ specific questionnaires. The PADQLQ showed good within-subject reliability and a small minimal important difference (0.33; 95% CI, 0.11 to 0.54 on a 7-point scale). CONCLUSIONS: The PADQLQ has good cross sectional and longitudinal validity, making it a potentially useful outcome measure in the evaluation of systemic treatments such as antihistamine medications and immunotherapy in children with multisystem allergic disease. PMID- 12642828 TI - Relations among asthma knowledge, treatment adherence, and outcome. AB - BACKGROUND: Asthma knowledge is frequently assumed to be a prerequisite for optimal asthma treatment. However, the validity of existing asthma knowledge questionnaires has not been rigorously examined, and no contemporary measure of asthma knowledge has received widespread acceptance. OBJECTIVE: To construct and examine the psychometric properties of an asthma knowledge instrument, and its association with demographic and psychosocial variables, asthma medication adherence, and treatment outcome. METHODS: A 25-item Asthma Knowledge Questionnaire was developed with input from national pediatric asthma experts. Parents of 155 children with asthma completed the Asthma Knowledge Questionnaire as well as demographic, family functioning, and home environment measures. Asthma outcomes and adherence with inhaled medication was measured across 12 months. RESULTS: Despite the many steps taken to develop a strong measure of asthma knowledge, reliability was relatively poor. There was also no association between asthma knowledge and treatment adherence or outcomes. Furthermore, asthma knowledge was not a unidimensional construct and was not a simple function of education. CONCLUSIONS: Findings from this study, in combination with previous studies of asthma knowledge questionnaires, suggest that the construction of a simple self-report asthma knowledge instrument for use as a primary outcome measure demonstrating mastery of asthma self-management skills may not be achievable. PMID- 12642829 TI - Inhaled corticosteroids and allergy specialty care reduce emergency hospital use for asthma. AB - BACKGROUND: The interrelationships between optimal inhaled corticosteroid (IC) therapy, allergy specialist care, and reduced emergency hospital care for asthma have not been well defined. OBJECTIVE: We sought to evaluate the independent effectiveness of various levels of IC dispensing and allergy specialist care in reducing subsequent emergency asthma hospital use. METHODS: Asthmatic patients (n = 9608) aged 3 to 64 years were identified from an electronic database of a large health maintenance organization. The outcome was any year 2000 asthma hospitalization or emergency department visit. The main predictors were at least one allergy department visit and the number of IC canisters dispensed in 1999. Analyses were adjusted for age, sex, insurance type, and asthma severity (1999 emergency asthma hospital use, beta-agonist use, and oral corticosteroid use). RESULTS: Dispensing of 7 or more canisters of ICs (odds ratio [OR], 0.64; 95% CI, 0.43-0.94) and allergy care (OR, 0.73; 95% CI, 0.55-0.97) were associated with reduced subsequent emergency asthma hospital use. More patients with allergy specialist care than those without such care received 7 or more dispensations of ICs (24.7% vs 8.3%, P <.001). When 7 or more dispensations of ICs and allergy specialist care were simultaneously included in an adjusted model, both ICs (OR, 0.68; 95% CI, 0.46-1.00) and allergy care (OR, 0.77; 95% CI, 0.58-1.02) were independently associated with a lower risk of year 2000 emergency asthma hospital care, although significance was borderline. CONCLUSION: Allergy care reduces emergency hospital use for asthma by increasing use of ICs but probably also has an independent effect. PMID- 12642830 TI - Adolescents with atopic disorders have an attenuated cortisol response to laboratory stress. AB - BACKGROUND: Patients with allergic disorders have been noted to have variations in cortisol patterns under natural conditions as well as a differential cortisol response to stress. OBJECTIVE: The main goal of this study was to examine hypothalamic-pituitary-adrenal (HPA) axis differences in atopic adolescents. METHODS: Subjects were a community sample of 202 adolescents (52% male; mean age, 16.2 years). Atopic status was determined by skin testing and clinical history. Saliva samples for cortisol assay were obtained 4 times during a "typical day" and at 4 time points during laboratory procedures. RESULTS: One third of the sample (33%) had a clinical atopic disorder, primarily allergic rhinitis; 39% had positive skin test results without clinical symptoms; and 27% had no signs or symptoms of allergic disorders. There were no significant effects of atopic status on home cortisol patterns. Presence of clinical atopy was significantly (P <.05) associated with lower cortisol levels in response to laboratory stressors. CONCLUSIONS: Adolescents with a history of atopic illnesses had an attenuated cortisol response to the stress of laboratory procedures compared with adolescents with positive skin test results alone or nonatopic adolescents. An attenuated cortisol response to a stressor may help us understand the link between stress and exacerbation of atopic illness. PMID- 12642831 TI - Clara cell protein 16 (CC16) gene polymorphism influences the degree of airway responsiveness in asthmatic children. AB - BACKGROUND: Several studies have indicated linkage of chromosome 11q12-13 to asthma and associated traits. Among other candidate genes, the Clara cell protein 16 (CC16) gene maps to this region. CC16 is expressed in the bronchial epithelium and exhibits potent anti-inflammatory properties. A single-nucleotide polymorphism (SNP) in the CC16 gene (A38G) was previously associated with asthma. OBJECTIVE: We evaluated the role of the CC16 SNP in pediatric asthma and asthma severity in 2 German study populations. METHODS: The German Multicenter Allergy Study (MAS) cohort (n = 872, 94 asthmatic patients) and 112 allergic asthmatic children recruited in Freiburg, Germany, were included in the present study. Histamine provocations were performed at the age of 7 years in the MAS cohort to determine bronchial hyperreactivity; in the Freiburg study population a standardized exercise-induced decrease in FEV1 was evaluated. For genotyping, melting-curve analysis and restriction enzyme digestion were applied. RESULTS: No association of the CC16*38A allele with asthma could be observed in either study population. However, in asthmatic subjects (MAS cohort) PC(20)FEV(1) values were significantly lower in individuals homozygous or heterozygous for the CC16*38A allele compared with those in subjects with the CC16*38GG genotype (P <.05 and P <.03, respectively). Similarly, allergic asthmatic patients in the Freiburg cohort showed a significantly greater decrease in FEV1 after exercise when homozygous for the CC16*38A allele compared with that seen in asthmatic patients with the *38AG or *38GG genotype (P <.04 and P =.006, respectively). CONCLUSION: We conclude that the CC16*A38G SNP influences bronchial hyperreactivity and might be a genetic determinant of asthma severity in German children. PMID- 12642832 TI - Differential expression of thymus- and activation-regulated chemokine (CCL17) and macrophage-derived chemokine (CCL22) by human fibroblasts from cornea, skin, and lung. AB - BACKGROUND: Allergic diseases of the ocular surface, skin, and lung are triggered by T(H)2 cells, which are recruited by thymus- and activation-regulated chemokine (TARC; CCL17) and macrophage-derived chemokine (MDC; CCL22). Resident fibroblasts are thought to contribute to inflammatory cell infiltration through chemokine production. OBJECTIVE: We sought to provide insight into the clinical differences apparent among these allergic diseases of the eye, skin, and lung, and we compared the abilities of corneal, dermal, and lung fibroblasts to produce TARC and MDC. METHODS: The amounts of chemokines released into the culture supernatant were determined by means of ELISA, and the intracellular abundance of chemokine mRNAs was quantitated by means of reverse transcription and real-time PCR analysis. RESULTS: Neither TNF-alpha, IFN-gamma, IL-4, nor IL-13 alone induced the release of TARC from or affected the amount of TARC mRNA in corneal, dermal, or lung fibroblasts. The combination of TNF-alpha with either IL-4 or IL-13, however, markedly increased both TARC release and the abundance of TARC mRNA in corneal and dermal fibroblasts, but not in lung fibroblasts. Neither MDC release nor MDC mRNA was detected in any of the 3 types of fibroblasts stimulated with any of the cytokines examined. CONCLUSION: These results indicate that cytokine regulation of TARC expression differs among fibroblasts derived from the cornea, skin, or lung. Corneal and dermal fibroblasts might thus be important sources of TARC during allergic inflammation. PMID- 12642833 TI - Sequence variation in the promoter region of the cholinergic receptor muscarinic 3 gene and asthma and atopy. AB - BACKGROUND: Muscarinic acetylcholine receptors are members of the superfamily of G protein-coupled, 7 transmembrane- spanning proteins. They are important in the development of airway hyperresponsiveness. In the lung the M3 receptor, encoded by the cholinergic receptor muscarinic 3 gene, is present in airway smooth muscle and mediates smooth muscle contraction. OBJECTIVE: We considered the cholinergic receptor muscarinic 3 gene as a possible candidate gene for bronchial asthma and initiated studies to identify polymorphisms in the promoter region. METHOD: We identified 4 single-nucleotide polymorphisms (-708A/G, -627G/C, -513C/A, and 492C/T) and 2 short tandem repeat polymorphisms, a tetranucleotide (CTTT)12-20 and a dinucleotide (GT)6-19 repeat. RESULTS: None of the identified single nucleotide polymorphisms were significantly more frequent in asthmatic patients (n = 76) compared with in healthy control subjects (n = 81). Furthermore, there was no evidence for nonrandom transmission of short tandem repeat polymorphism haplotypes to individuals with asthma or bronchial hyperresponsiveness (P >.50) in a large Hutterite pedigree. However, there was significant nonrandom transmission of haplotypes to individuals with skin test reactivity to cockroach allergens (global transmission disequilibrium test: chi2 = 38.55, P =.013). CONCLUSIONS: These results suggest a possible role for this gene in atopic disorders. PMID- 12642834 TI - The effect of hydrolyzed cow's milk formula for allergy prevention in the first year of life: the German Infant Nutritional Intervention Study, a randomized double-blind trial. AB - BACKGROUND: The potential of extensively or partially hydrolyzed formulas to reduce the risks for allergies is controversial. OBJECTIVE: We sought to assess the preventive effect of differently hydrolyzed formulas compared with cow's milk formula (CMF) in high-risk infants. METHODS: Between 1995 and 1998, 2252 infants with a hereditary risk for atopy were enrolled in the German Infant Nutritional Intervention Study and randomly assigned at birth to one of 4 blinded formulas: CMF, partially hydrolyzed whey formula, extensively hydrolyzed whey formula, and extensively hydrolyzed casein formula (eHF-C). The primary end point at 1 year of age was the presence of allergic manifestation, which was defined as atopic dermatitis (AD), gastrointestinal manifestation of food allergy, allergic urticaria, or a combination of these factors. RESULTS: At 12 months per protocol, analysis was performed on 945 infants exposed to study formula: 304 (13.5%) infants had left the study, 138 (6.1%) infants were excluded because of noncompliance, and 865 infants were exclusively breast-fed the first 4 months of life. The incidence of allergic manifestation was significantly reduced by using eHF-C compared with CMF (9% vs 16%; adjusted OR, 0.51; 95% CI, 0.28-0.92), and the incidence of AD was significantly reduced by using eHF-C (OR, 0.42; 95% CI, 0.22-0.79) and partially hydrolyzed whey formula (OR, 0.56; 95% CI, 0.32-0.99). Family history of AD was a significant risk factor and modified the preventive effect of the hydrolysates. CONCLUSIONS: Prevention of allergic diseases in the first year of life is feasible by means of dietary intervention but influenced by family history of AD. The preventive effect of each hydrolyzed formula needs to be clinically evaluated. PMID- 12642835 TI - The effect of neonatal BCG vaccination on atopy and asthma at age 7 to 14 years: an historical cohort study in a community with a very low prevalence of tuberculosis infection and a high prevalence of atopic disease. AB - BACKGROUND: There are conflicting reports on the effect of BCG vaccination on the subsequent development of atopy and asthma. There are no data on the effects of neonatal BCG vaccination on cytokine responses of lymphocytes that are exposed in vitro to allergens. OBJECTIVES: We sought to test the hypothesis that neonatal BCG vaccination or, alternatively, evidence of an immunologic memory of this vaccination is associated with a reduced prevalence of allergic sensitization, asthma, eczema, and hay fever during childhood. METHODS: An historical cohort study was conducted among 7- to 14-year-old children who were born in 2 districts in Sydney, Australia, and whose mothers were born in southeast Asia. One district had routinely administered BCG vaccination to infants born to overseas-born mothers and the other had not. Eligible subjects were identified from birth registers. Consenting subjects completed questionnaires, performed spirometric and airway hyperresponsiveness testing, and had allergen skin prick testing and tuberculin skin testing. Blood was collected to measure total serum IgE levels and for in vitro lymphocyte culture in the presence of an extract of house dust mite, the dominant allergen in this region, and purified protein derivative of Mycobacterium tuberculosis (tuberculin). IL-4, IL-5, IL-10, and IFN-gamma were measured in the culture supernatant. RESULTS: The cohort included 309 BCG vaccinated subjects and 442 non-BCG-vaccinated subjects. BCG-vaccinated subjects did not have a lower rate of allergic sensitization than nonvaccinated subjects. However, among the subgroup of subjects with a family history of rhinitis or eczema, BCG vaccination was associated with a lower prevalence of current asthma (defined as recent wheezing plus airway hyperresponsiveness; relative risk, 0.46; 95% CI, 0.22-0.95). BCG vaccination was also associated with lower levels of allergen-stimulated IL-10 production in vitro. Among the BCG-vaccinated subjects, the 44 (14.3%) who had tuberculin skin test reaction sizes of 5 mm or greater and the 31 (18.3%) who demonstrated an in vitro IFN-gamma response to purified protein derivative of M tuberculosis did not have lower rates of allergic sensitization and, overall, did not have a lower prevalence of allergic disease than tuberculin skin test or IFN-gamma nonreactors. CONCLUSION: We conclude that neonatal BCG vaccination has an effect on T-cell allergen responsiveness 7 to 14 years after vaccination and that among a subgroup of subjects with an inherited predisposition to allergic disease, this is associated with clinically relevant beneficial effects. The findings of this study encourage the view that external influences on the immune system in the neonatal period have consequences that extend into later childhood and influence the expression of asthma. Genetic factors are likely to modify the effect of those external factors. PMID- 12642836 TI - Upregulation of B7 molecules (CD80 and CD86) and exacerbated eosinophilic pulmonary inflammatory response in mice lacking the IFN-beta gene. AB - BACKGROUND: IFN-beta has been shown to be effective as therapy for multiple sclerosis. Some reports attributed its beneficial effects to the capacity to induce a T(H)2 response. However, other studies have suggested that endogenous type I IFN might downregulate the allergic response in mice. OBJECTIVE: We sought to define the differential role of endogenous IFN-beta in controlling the development of allergic inflammation. METHODS: We assessed whether deletion of the gene encoding IFN-beta (IFNB) with knockout mice participated in the development of allergic response in ovalbumin (OVA)-sensitized and OVA-challenged mice. RESULTS: OVA-sensitized and OVA-challenged mice with lack of the IFNB gene had more severe pulmonary inflammation with increased lung local response, including IL-4, IL-5, IL-13, IgE, eosinophilia, and goblet cells, than their litter mates (IFN-beta+/-), whereas no differences were observed in regard to local levels of IFN-gamma. Moreover, systemic response with IgE production is also enhanced. Lack of IFN-beta also results in significantly higher antigen specific T cells, with higher levels of IL-4, IL-5, and IL-13, whereas no significant differences in IFN-gamma response could be observed. We have also detected a higher ratio of CD4+/CD8+ T cells and increased expression of B7.1/B7.2 on B cells and antigen-presenting cells in IFNB knockout mice. CONCLUSIONS: These results demonstrate that IFN-beta plays an important role in immunoregulation of allergic response in mice. The stronger pulmonary inflammation could be a consequence of significantly expanded antigen-specific CD4+ T(H)2 cells as a result of efficient antigen presentation by antigen presenting cells and hence increased production of IgE by B cells. PMID- 12642837 TI - The effects of IL-5 on airway physiology and inflammation in rats. AB - BACKGROUND: There is evidence that the cytokine IL-5 is a prominent feature of airway inflammation in asthma. OBJECTIVE: The aim of this study was to determine whether exogenous IL-5 could cause changes in lung physiology, the early and late airway response after antigen challenge, and airway inflammation in rats that do not have a propensity to develop these changes after sensitization and challenge. METHOD AND RESULTS: Intratracheal administration of IL-5 to ovalbumin sensitized Brown Norway SSN rats increased the airway responsiveness to methacholine (AHR) 20 hours after administration of IL-5 at the same time as an increase in neutrophils occurred in the lung lavage. This effect was dose dependent and was not caused by endotoxin. Concurrent intratracheal administration of 50 ng of anti IL-5 monoclonal antibody with 10 microg of recombinant human IL-5 decreased the AHR and neutrophil influx. Pretreatment with 3 microg of IL-5 had no effect on the early and late airway response or on AHR after ovalbumin challenge. However, IL-5 increased lung re-sistance 20 hours after antigen challenge. Although total lung cells and differential counts did not differ significantly 8 hours after antigen challenge, the blood lymphocyte CD4/CD8 ratio decreased in IL-5 pretreated rats (P <.05). In addition, in situ hybridization showed a significant increase in cells within the airway wall expressing IL-4 and IL-5 mRNA in IL-5 treated/challenged rats compared to controls (P <.05). CONCLUSION: The intratracheal administration of IL-5 causes only part of the physiologic changes that are associated with asthma. Other factors are necessary to obtain the complete asthma phenotype. PMID- 12642838 TI - The antiallergic drug oxatomide promotes human eosinophil apoptosis and suppresses IL-5-induced eosinophil survival. AB - BACKGROUND: Eosinophils accumulated in sites of allergic inflammation are thought to play a crucial role in the pathogenesis of allergic disorders including asthma, allergic rhinitis, and atopic dermatitis, and tissue eosinophilia is attributable to increased eosinophil survival or decreased eosinophil apoptosis. OBJECTIVE: Effects of the antiallergic, histamine H1 blocker oxatomide on viability and apoptosis of eosinophils isolated from the peripheral blood of atopic subjects were studied. METHODS: Eosinophil viability and apoptosis were evaluated by using a colorimetric assay and annexin V-labeling, caspase-3 activity, and DNA fragmentation assay. RESULTS: The viability of eosinophils increased in the presence of IL-5 (10 ng/mL), confirming that IL-5 prolongs eosinophil survival in vitro. Application of oxatomide at concentrations over 20 micromol/L for 24 hours decreased the IL-5-induced enhancement of eosinophil viability. Double staining of the cells with annexin V and propidium iodide showed that deprivation of IL-5 promoted spontaneous eosinophil apoptosis and that oxatomide facilitated apoptosis and suppressed the prolongation of eosinophil survival stimulated by IL-5. In the absence of IL-5, approximately 71% and 96% of eosinophils after 24 and 48 hours, respectively, underwent spontaneous apoptosis. IL-5 decreased the rate of eosinophil apoptosis to 38% and 52% after 24 and 48 hours, respectively. Oxatomide increased eosinophil apoptosis in a concentration-dependent manner in the presence of IL-5. Furthermore, oxatomide increased caspase-3 activity and DNA fragmentation. CONCLUSION: We demonstrated that oxatomide possesses a novel therapeutic effect of apoptosis promotion on eosinophils and prevents the antiapoptotic effects of IL-5, suggesting that oxatomide may contribute to resolution of tissue eosinophilia in allergic inflammation. PMID- 12642839 TI - Long-term assessment of T-cell populations in DiGeorge syndrome. AB - BACKGROUND: Patients with DiGeorge syndrome present with a broad range of T-cell deficiency. Partial DiGeorge syndrome (pDGS) is a preferred designation for patients with detectable T-cell function. Among immunology experts, there is no uniform opinion on the necessity of T-cell precautions for pDGS patients. Few studies have addressed the natural course of their immune function over time. OBJECTIVE: The objective of this study was to describe the natural history of immune parameters in pDGS. METHODS: We reviewed the medical records of 45 pDGS patients. Peripheral blood T-cell subsets counts and percentages were recorded at 1, 6, 12, 18, 24, 30, 48, 60, 72, 96, and 120 months of age, and the rates of change of T-cell measurements over the follow-up period (slopes) were calculated for each individual. Humoral immunity was evaluated by quantification of immunoglobulins and by testing antibody titers to recall antigens. RESULTS: T cell subsets counts from pDGS patients were generally lower than those of age matched normal populations but were not severely depressed (ie, CD4+ T-cell percentage less than 15%). The median of the slopes for CD3+, CD4+, and CD8+ T cell percentages were -0.7%, -0.8%, and -0.1%/month, respectively, in the first year of age and 0.1%/month for each subpopulation from 12 to 120 months of age. Lymphoproliferative responses to phytohemagglutinin were adequate at all ages. Immunoglobulin deficiencies or inadequate production of specific antibodies were not detected. CONCLUSIONS: In our pDGS patient cohort, a significant deterioration of T-cell number or function did not occur over time. Clinical implications of this finding include the possibility of discontinuing T-cell deficiency precautions and frequency of reevaluations of pDGS patients with stable and adequate immune function. PMID- 12642840 TI - Modulation of allergic response in nasal mucosa by antisense oligodeoxynucleotides for IL-4. AB - BACKGROUND: The cytokine IL-4 is highly involved in T(H)2 inflammation, such as that seen in allergic rhinitis. IL-4 can induce IgE synthesis and eotaxin. Recent studies have shown that stimulation of allergic nasal tissue with ragweed allergen can induce local IL-4 mRNA production. OBJECTIVE: We set out to determine whether IL-4 antisense phosphorothioate-modified oligodeoxynucleotides (PS-ODNs) could inhibit IL-4 production and downstream events of IL-4. METHODS: Nasal mucosa biopsy specimens were obtained from patients with seasonal ragweed allergic rhinitis out of season, incubated ex vivo with or without PS-ODNs, and challenged with ragweed. FITC-labeled oligonucleotides were used to determine tissue uptake. By using immunocytochemistry, IL-4-, IL-13-, eotaxin 1-, and IFN gamma-producing cells were enumerated, and by using in situ hybridization, epsilon germline transcript RNA- and IL-4 mRNA-positive cells were examined. RESULTS: The antisense PS-ODN was taken up by the tissue, and preincubation of the tissue with the IL-4 antisense PS-ODN caused a decrease in allergen-induced IL-4 mRNA and a decrease in the amount of IL-4 immunoreactivity (n = 7, P <.001). PS-ODNs had inhibitory effects on allergen-induced epsilon germline transcript RNA expression (n = 7, P <.001) and mucosa eotaxin 1 immunoreactivity (n = 7, P <.05). In contrast, the PS-ODNs increased the amount of IFN-gamma immunoreactivity (n = 7, P <.05), suggesting a nonspecific mechanism for reduced synthesis of IgE and eotaxin. CONCLUSIONS: Our results show that the IL-4 antisense PS-ODN effectively inhibits IL-4, IgE synthesis, and eotaxin, principal mediators of allergic inflammation, suggesting that PS-ODNs might offer a possible topical treatment for allergic rhinitis. PMID- 12642841 TI - Differences in fecal microflora between patients with atopic dermatitis and healthy control subjects. AB - BACKGROUND: The prevalence of allergic diseases, such as atopic dermatitis (AD), has been increasing. However, few investigations have been made of the intestinal microflora in Japanese patients with AD. OBJECTIVE: The purpose of this study was to determine the differences in microflora, fecal serum IgA concentrations, and skin IgA contents between patients with AD and healthy control subjects. METHODS: This trial was conducted as a case-control study using 30 minor patients with AD and age- and sex-matched healthy control subjects (n = 68). One week after a questionnaire was administered, fecal specimens and 24-hour skin secretion specimens were collected from all subjects. Fecal microflora, fecal IgA concentrations, and IgA contents on the skin surface were analyzed. RESULTS: The counts of Bifidobacterium (in log10 colony-forming units per gram) were significantly lower in patients with AD than in healthy control subjects (9.75 +/ 0.68 vs 10.10 +/- 0.50 log(10) colony-forming units/g, P <.05). In particular, percentages of Bifidobacterium were significantly lower in patients with severe skin symptoms than in those with mild skin symptoms (40% +/- 6% vs 19% +/- 6%, P <.05). In addition, the frequency of occurrence of Staphylococcus was significantly higher in patients with AD than in healthy control subjects (83% vs 59%, P <.05). There were no significant differences in fecal IgA content or IgA content on the skin between the 2 groups. CONCLUSION: Patients with AD had lower counts of Bifidobacterium than healthy control subjects, and the frequency of Staphylococcus was higher in patients with AD than in control subjects. Disorder of the intestinal microflora might play a role in the onset of AD and the aggravation of skin symptoms. PMID- 12642842 TI - Increased serum cutaneous T cell-attracting chemokine (CCL27) levels in patients with atopic dermatitis and psoriasis vulgaris. AB - BACKGROUND: Both atopic dermatitis (AD) and psoriasis vulgaris (PsV) are characterized as chronic and relapsing inflammatory skin diseases associated with various immunologic abnormalities. Cutaneous T cell-attracting chemokine (CTACK; CCL27) is a member of the CC chemokine family and a functional ligand for CC chemokine receptor 10. It is selectively expressed in skin and attracts CC chemokine receptor 10-expressing skin-homing memory T cells. The epidermal keratinocyte is a main source of CTACK, suggesting the involvement of various inflammatory skin diseases. OBJECTIVE: The purpose of this investigation was to clarify whether CTACK produced by keratinocytes is detected in the sera of patients with AD and PsV and to examine the correlation between the serum CTACK levels and disease activity of patients with AD and PsV. METHODS: We measured the serum CTACK levels in 50 patients with AD, 30 patients with PsV, and 22 healthy control subjects. We also divided 50 patients with AD into 3 groups (ie, those with mild, moderate, and severe disease) and compared them among 3 categories. Moreover, we compared the serum CTACK levels of patients with AD and PsV with clinical or laboratory data. Immunohistochemical staining of CTACK and IFN induced protein of 10 kd (IP-10; CXCL10) was performed on the lesional skin of patients with AD and PsV. RESULTS: The serum CTACK levels in patients with AD and PsV were significantly higher than those in healthy control subjects. The serum CTACK levels in patients with AD significantly correlated with scoring atopic dermatitis (SCORAD) scores, serum soluble IL-2 receptor levels, serum soluble E selectin levels, serum thymus and activation-regulated chemokine levels, and serum macrophage-derived chemokine levels. Serum CTACK levels in patients with PsV significantly correlated with the serum IP-10 levels but not with the Psoriasis Area and Severity Index score. Immunohistochemical staining showed CTACK was strongly expressed in lesional ke-ratinocytes of patients with AD and PsV, whereas IP-10 was strongly expressed in lesional keratinocytes of patients with PsV and focally in those with AD. CONCLUSION: These results suggest that CTACK might be one of the important chemokines for the pathogenesis of AD and PsV. PMID- 12642843 TI - Actigraphy assessment of sleep disturbance in patients with atopic dermatitis: an objective life quality measure. AB - BACKGROUND: Patients with atopic dermatitis (AD) frequently report compromised quality of life because of disturbed sleep and daytime fatigue secondary to their skin disease, but few studies provide objective measurement of sleep change in this population. OBJECTIVE: The purpose of this investigation was to contrast subjective and objective measures of sleep quality in patients with AD. METHODS: Fourteen adult patients with AD and 14 adult control subjects with no skin disease wore actigraphs for 1 week and completed questionnaires about sleep, itch, and quality of life. RESULTS: As measured by self-report and actigraphy, the AD group slept more poorly and reported more daytime fatigue than the control group. Actigraphy alone was correlated with itch and quality of life and was able to discriminate movement during sleep, number of awakenings, minutes asleep, and minutes awake. CONCLUSIONS: Results from this study demonstrate that sleep is significantly compromised in patients with AD. Patients' perception of their sleep provides less detail and accuracy than actigraphy. The actigraph is an objective, unobtrusive measure of sleep at home in patients with skin disease and can provide an important outcome measure in clinical trials. PMID- 12642844 TI - An extensive study of human IgE cross-reactivity of Blo t 5 and Der p 5. AB - BACKGROUND: Dual sensitization by Blomia tropicalis and Dermatophagoides pteronyssinus mites is common in tropical and subtropical countries. The human IgE cross-reactivity between clinical important group 5 allergens, Blo t 5 and Der p 5, remains controversial. OBJECTIVE: This study was undertaken to assess the levels of the IgE cross-reactivity between Blo t 5 and Der p 5 by using sera from a large cohort of asthmatic children in subtropical and tropical countries. METHODS: Purified recombinant Blo t 5 and Der p 5 were produced in Pichia pastoris and tested against sera from 195 asthmatic children. The IgE cross reactivity was examined by direct, inhibitory and competitive human IgE enzyme linked immunosorbent assay as well as skin prick tests. RESULTS: The Blo t 5 IgE responses were 91.8% (134 of 146) and 73.5% (36 of 49) for Taiwanese and Malaysian sera, respectively. The Blo t 5 specific IgE titers were significantly higher than those of Der p 5 (P <.02). The correlation of IgE reactivity between Blo t 5 and Der p 5 was low, and only limited cross-reactivity was observed. This was further confirmed by the dose-response inhibition studies. Skin prick tests performed on asthmatic children in Thailand also showed differential IgE response to Blo t 5 and Der p 5. CONCLUSION: By using a large panel of asthmatic sera and a combination of in vitro and in vivo assays, the major allergen of B tropicalis in tropical and subtropical regions, Blo t 5, exhibits low levels of IgE cross reactivity with homologous Der p 5. These findings suggest that highly specific clinical reagents are necessary for precise diagnosis and immunotherapeutic treatment of sensitization to group 5 mite allergens. PMID- 12642845 TI - In vivo sensitization to purified Hevea brasiliensis proteins in health care workers sensitized to natural rubber latex. AB - BACKGROUND: Thirteen proteins of natural rubber latex (Hevea brasiliensis) known to bind human IgE have been isolated and characterized as Hev b allergens. However, the in vivo importance of native Hev b allergens has not been defined in health care workers (HCWs) with natural rubber latex (NRL) allergy. OBJECTIVES: The principal aim of this study was to identify the major in vivo Hev b allergens in HCWs with NRL allergy confirmed by percutaneous sensitivity to nonammoniated latex (NAL). METHODS: Skin prick testing was performed with 7 (native) proteins purified from NAL (Hev b 1, 2, 3, 4, 6.01, 7.01, and a newly described Hev b 13) and recombinant Hev b 5 in 62 HCWs with histories of NRL allergy (group 1) confirmed by percutaneous reactivity to NAL and in 49 atopic HCWs without NRL allergy (group 2). Serial 10-fold concentrations of Hev b proteins (5 x 10(-5) microg/mL to 50 microg/mL) were tested; serum samples of subjects were assayed for serum specific IgE by immunoassays. RESULTS: Hev b 2, Hev b 5, Hev b 6.01, and Hev b 13 produced skin reactions in more than 60% of group 1 subjects, with Hev b 1, 3, 4, and 7.01 eliciting reactions in less than 50%. Only 1 of 49 group 2 workers reacted to a single Hev b antigen (Hev b 13). Specificity of 7 Hev b allergens was 100% and 98% for Hev b 13 in identifying workers with confirmed NRL allergy. Specific IgE by AlaSTAT and CAP immunoassays was elevated in 40 of 60 (67%) and 33 of 62 (53%) of NAL-reactive workers and produced false-positive test results in 4 of 49 (8%) and 3 of 48 (6%) group 2 subjects, respectively. CONCLUSION: Hev b 2, 5, and 6.01 are major in vivo allergens and Hev b 13 is a new major in vivo allergen among HCWs with allergy to NRL. PMID- 12642846 TI - Efficacy and safety of desloratadine in the treatment of perennial allergic rhinitis. AB - BACKGROUND: The evidence base for the use of H1-antihistamines in the treatment of perennial allergic rhinitis is considerably smaller than it is in the treatment of seasonal allergic rhinitis. OBJECTIVE: We hypothesized that desloratadine, a new, nonsedating selective H1-antihistamine, would be efficacious and safe in the treatment of perennial allergic rhinitis. METHODS: In a multicenter, randomized, placebo-controlled, double-blind, parallel-group study, 676 patients with symptomatic perennial allergic rhinitis were randomly assigned to 4 weeks of treatment with either 5 mg of desloratadine once daily or placebo. Efficacy was assessed by using a morning-evening instantaneous total symptom score (TSS), which was composed of scores for 4 individual nasal symptoms (rhinorrhea, itching, sneezing, and postnasal drip) and 3 individual nonnasal symptoms (itching eyes, watering eyes, and itching of the ears or palate). Secondary outcome measures included a morning-evening reflective TSS, total nasal and nonnasal symptoms scores, and individual symptom scores. Safety evaluations, including 12-lead electrocardiograms, were performed. RESULTS: Six hundred thirty four patients completed the study. Desloratadine consistently diminished perennial allergic rhinitis symptoms, reducing the morning-evening instantaneous TSS (P =.005), the morning-evening reflective TSS (P =.007), the morning-evening reflective total nonnasal score (P =.023), and the individual nasal symptom scores for rhinorrhea, nasal itching, sneezing, and postnasal drip/drainage (P =.05 to P =.013) during weeks 1 through 4. Improvement in symptoms was observed after the first dose. Dropouts, and the type and frequency of adverse events (headache, viral infection, pharyngitis, and upper respiratory tract infection), were similar in both treatment groups. No clinically significant changes in QTc intervals were observed. CONCLUSIONS: Desloratadine rapidly and safely reduced the symptoms of perennial allergic rhinitis, and its efficacy did not diminish during 4 weeks of treatment. PMID- 12642847 TI - Acute and subchronic effects of levocetirizine and diphenhydramine on memory functioning, psychomotor performance, and mood. AB - BACKGROUND: Central nervous system adverse effects, such as sedation, often accompany the use of first-generation antihistamines. These effects might interfere with memory functioning and psychomotor performance. Levocetirizine was recently introduced as a new antihistamine said to be free from sedative effects. OBJECTIVE: We sought to investigate the effects of levocetirizine (5 mg), diphenhydramine (50 mg), and placebo on memory and psychomotor performance after acute (day 1) and subchronic (day 4) daily administration in 48 healthy volunteers (24 men and 24 women). METHODS: This study was a double-blind, placebo controlled, randomized clinical trial. Treatments were administrated on days 1, 2, 3, and 4, 3 hours before the start of the laboratory test battery (performed on days 1 and 4), comprising a word-learning test, the Sternberg Memory Scanning Test, a tracking test (easy and hard version), and a divided attention test (tracking and memory scanning simultaneously). Statistical analyses were performed separately for days 1 and 4 by using analysis of variance. RESULTS: On day 1, diphenhydramine significantly impaired tracking performance (easy: F(1,90) = 25.9, P <.0001; hard: F(1,90) = 20.5, P <.0001) and divided attention (tracking: F(1,90) = 23.8, P <.0001; memory scanning: F(1,90) = 22.0, P <.0001). Results on word-learning tests and Sternberg Memory Scanning Tests were not significantly impaired. On day 4, the effects of diphenhydramine did not reach significance. In contrast, on both days 1 and 4, levocetirizine did not significantly impair laboratory test performance. CONCLUSION: The results show that memory, attention, and tracking performance are unaffected after acute and subchronic administration of levocetirizine (5 mg), whereas diphenhydramine (50 mg) significantly affected divided attention and tracking after acute administration. PMID- 12642848 TI - Recombinant Pru p 3 and natural Pru p 3, a major peach allergen, show equivalent immunologic reactivity: a new tool for the diagnosis of fruit allergy. AB - BACKGROUND: The peach lipid transfer protein Pru p 3 has been identified as a major allergen from this fruit. Homologous cross-reactive allergens have been found in several plant foods and pollens. Recombinant Pru p 3 has been recently produced in the yeast Pichia pastoris. OBJECTIVE: We sought to evaluate the potential role of recombinant Pru p 3 as a novel tool for the diagnosis of fruit allergy. METHODS: Circular dichroism analysis was used to compare the protein folding of natural Pru p 3 and recombinant Pru p 3. IgE binding by both molecular forms was quantified by means of ELISA and ELISA inhibition assays, and their biologic activity was estimated by using basophil activation, histamine release, and sulphidoleukotriene production tests. Individual sera or blood samples from patients with peach allergy (up to 17) were used in the assays. RESULTS: A nearly identical circular dichroism spectra was shown by using natural Pru p 3 and recombinant Pru p 3, indicating that both protein forms are similarly folded. No difference was detected in the IgE-binding capacity of the 2 mo-lecular versions. Basophil activation and induction of sulphidoleukotriene production were positive in 9 of 10 patients, and histamine release was induced in at least half of the patients, with similar effects of the natural and recombinant forms in the 3 assays. CONCLUSION: Recombinant Pru p 3 shows a strong immunologic activity equivalent to that of its natural counterpart, and therefore it can be a useful tool for diagnosis (and future immunotherapy) of fruit allergy. PMID- 12642849 TI - Profilin is a relevant melon allergen susceptible to pepsin digestion in patients with oral allergy syndrome. AB - BACKGROUND: Melon allergy has been documented by means of double-blind, placebo controlled food challenges. The most common clinical feature associated with melon allergy is oral allergy syndrome (OAS). However, no relevant allergens of melon have been identified to date. OBJECTIVE: We sought to identify melon allergens and analyze their digestibility in human saliva and simulated gastric fluid (SGF) to provide a rationale for the OAS. METHODS: Melon, zucchini, cucumber, and watermelon allergens were identified by means of IgE immunoblotting of sera from 21 patients with OAS after melon ingestion confirmed by means of double-blind, placebo-controlled food challenge. Further characterization was performed with rabbit antisera against sunflower pollen profilin and anticomplex glycans. Lability of allergens was assayed by incubation of melon extract in human saliva and SGF. RESULTS: Several IgE-binding components between 15 and 60 kd and a main reactive band of 13 kd were detected in melon extract with the pooled sera from patients with melon allergy. As in melon, 13-kd components of zucchini, cucumber, and watermelon extracts were strongly recognized by the IgE antibodies of the patients with melon allergy and were identified as profilins. Putative cross-reacting carbohydrate determinants were also detected. Sera from 71% of patients recognized the melon profilin, and therefore profilin is considered a major allergen. Melon allergens were unaffected by crude human saliva. In contrast, most melon proteins, predominantly the 13-kd component (profilin), were quickly digested in the SGF. CONCLUSION: In patients with OAS, a 13-kd protein identified as a profilin is a major melon allergen highly susceptible to pepsin digestion. PMID- 12642850 TI - Monitoring peanut allergen in food products by measuring Ara h 1. AB - BACKGROUND: Peanut allergy is an important health problem in the United States, affecting approximately 0.6% of children. Inadvertent exposure to peanut is a risk factor for life-threatening food-induced anaphylaxis. OBJECTIVE: The purpose of this investigation was to develop an immunoassay for a major peanut allergen, Ara h 1, to detect peanut allergen in foods so that the risk of inadvertent exposure can be reduced. METHODS: A specific 2-site monoclonal antibody-based ELISA was developed to measure Ara h 1 in foods. The sensitivity of the assay was 30 ng/mL. Ara h 1 was measured in foods (n = 83) with or without peanut and in experiments to optimize allergen yield and to determine peanut contamination in spiked foods. RESULTS: Ara h 1 levels in food products ranged from less than 0.1 microg/g to 500 microg/g. Ara h 1 measured in ng/mL was transformed to microg/g for food products. Peanut butter contained the highest amounts of Ara h 1. Peanut extracts contained from 0.5 to 15 mg Ara h 1/g of peanut depending on the extraction conditions. Optimal extraction of Ara h 1 was obtained by using phosphate buffer with 1 mol/L NaCl and Tween at 60 degrees C. Ara h 1 was not always detected in presence of chocolate under the extraction conditions tested. Spiking experiments showed that the assay could detect approximately 0.1% Ara h 1 contamination of food with ground peanut. There was an excellent correlation between Ara h 1 levels and peanut content measured by using a commercial polyclonal antibody-based ELISA (r = 93, n = 31, P <.001). CONCLUSION: A new sensitive and specific monoclonal antibody-based ELISA was used to monitor Ara h 1 content in food products. This assay should be useful for monitoring peanut contamination in the food manufacturing and processing industry and in developing thresholds for sensitization or allergic reaction in persons with peanut allergy. PMID- 12642851 TI - Roxithromycin enhances lymphocyte apoptosis in Dermatophagoides-sensitive childhood asthma. PMID- 12642852 TI - Cow's milk-dependent, exercise-induced anaphylaxis: case report of a patient with previous allergy to cow's milk. PMID- 12642853 TI - A prospective evaluation of the adequacy of peak inspiratory flow rates and validation of the Turbutester score in asthmatic individuals with "low" peak expiratory flow rates: implications for dry powder inhalers. PMID- 12642854 TI - Psychosomatic peanut allergy. PMID- 12642856 TI - Epidemiology of latex sensitization. PMID- 12642858 TI - Decongestant effects of antihistamines: a class effect? PMID- 12642859 TI - Conclusions of cost-effectiveness for an asthma intervention might not be warranted. PMID- 12642862 TI - Role of Smac in human leukaemic cell apoptosis and proliferation. AB - Smac (or DIABLO) is a recently identified, novel proapoptotic molecule, which is released from mitochondria into the cytosol during apoptosis. Smac functions by eliminating the caspase-inhibitory properties of the inhibitors of apoptosis proteins (IAP), particularly XIAP. In this study, we stably transfected both full length (FL) and mature (MT) Smac genes into the K562 and CEM leukaemic cell lines. Both FL and MT Smac transfectants increased the sensitivity of leukaemic cells to UV light-induced apoptosis and the activation of caspase-9 and caspase 3. Purified cytosol from the mature Smac transfectants, or the addition of human recombinant Smac protein or N-7 peptide into nontransfected cytosol, showed an increased sensitivity to cytochrome c-induced activation of caspase-3. The mature Smac enhanced the susceptibility of both K562 and CEM cells to TRAIL-induced apoptosis. Overexpression of the mature Smac protein also inhibited proliferation, as detected by reduced colony formation and Ki-67 expression in leukaemic cells. Cell cycle analysis revealed that Smac transfectants displayed significant G0/G1 arrest and reduction in 5-bromo-2'-deoxyuridine (BrdU) incorporation. Smac sensitized human acute myeloid leukaemia blasts to cytochrome c-induced activation of caspase-3. However, Smac failed to overcome Apaf-1 deficiency-mediated resistance to cytochrome c in primary leukaemic blasts. In summary, this study reveals that Smac/DIABLO exhibits a potential role in increasing apoptosis and suppressing proliferation in human leukaemic cells. Importantly, it also indicates that it is crucial to evaluate the levels of Apaf 1 and XIAP proteins in patient samples before using Smac peptide therapy in the treatment of human leukaemia. PMID- 12642863 TI - Consistent inactivation of p19(Arf) but not p15(Ink4b) in murine myeloid cells transformed in vivo by deregulated c-Myc. AB - Cyclin-dependent kinase inhibitors p16(INK4a) and p15(INK4b), encoded by the CDKN2A and B loci, play an important role in negative regulation of the cell cycle. Furthermore, p19(ARF) also encoded by the CDKN2A locus, has been shown to regulate positively the p53 pathway leading to growth arrest and apoptosis. All three genes have been inactivated in human tumors. In myeloid cells, p15(INK4b) mRNA is upregulated during cytokine-induced differentiation and/or growth arrest, and hypermethylation of the p15(INK4b) gene promoter region is a common event in acute myeloid leukemia. In the present study, we examined murine monocyte/macrophage tumors with deregulated c-myc for evidence of Ink4 gene inactivation. p15(Ink4b) mRNA and protein were detected in the majority of leukemias, and p16(Ink4a) mRNA and protein were highly expressed in two of them. pRb was in a hypophosphorylated state in most of the neoplasms indicating that the Cdk inhibitors that were expressed in the cells were functional. The observed expression of p15(Ink4b) is inconsistent with their proliferation state, although it might be expected to be expressed owing to the maturity of the cells. These data suggest, therefore, that deregulated c-Myc bypasses the pRb restriction point and cell cycle arrest in these tumors. An examination of p19(Arf) exons revealed deletions of the gene in up to 94% of the tumors. Since this gene shares exon 2 with p16(Ink4a), it is often difficult to determine which gene is the relevant tumor suppressor. However, the loss of only the p19(Arf)-specific exon 1 beta was observed in a tumor that had normal p16(Ink4a) protein expression. In addition, the p19(Arf)-specific exon was deleted in another tumor that expressed a functional chimeric protein, p15Ex1-p16Ex2-3; it was demonstrated here that this fusion protein is capable of inducing G1 arrest. These data overall supports the hypothesis that the critical inactivation event in these hematopoietic neoplasms is elimination of p19(Arf), and not Ink4 function. PMID- 12642864 TI - Distinct nuclear body components, PML and SMRT, regulate the trans-acting function of HTLV-1 Tax oncoprotein. AB - Several viruses target cellular promyelocytic leukemia (PML)-nuclear bodies (PML NBs) to induce their disruption, marked morphological changes in these structures or the relocation to PML-NB components to the cytoplasm of infected cells. PML conversely interferes with viral replication. We demonstrate that PML acts as a coactivator for the human T-cell leukemia virus type 1 (HTLV-1) Tax oncoprotein without direct binding. Tax was identified within interchromatin granule clusters (IGCs)/RNA splicing bodies (SBs), not PML-NBs; Tax expression did not affect PML NB formation. Moreover, PML and CBP/p300 cooperatively activated Tax-mediated HTLV-1-LTR-dependent gene expression. Interestingly, two PML mutants, PML-RAR and PMLDelta216-331, which fail to form PML-NBs, could also coactivate Tax-mediated trans-acting function but had no effect on retinoic acid receptor (RAR)- or p53 dependent gene expression. In contrast, SMRT (silencing mediator for retinoic acid and thyroid hormone receptors), a nuclear corepressor found within the matrix-associated deacetylase (MAD) nuclear body, relocalized into Tax-associated nuclear bodies upon coexpression with Tax. SMRT coactivated the trans-acting function of Tax through direct binding. Coexpression of SMRT and PML resulted in an additive activation of Tax trans-acting function. Thus, crosstalk between distinct nuclear bodies may control Tax function. PMID- 12642865 TI - UV-C-induced DNA damage leads to p53-dependent nuclear trafficking of PML. AB - The promyelocytic leukemia protein (PML) is a nuclear phosphoprotein that localizes to distinct domains in the nucleus, described as PML nuclear bodies (PML-NBs). Recent findings indicate that PML regulates the p53 response to oncogenic signals. Here, we define a p53-dependent role for PML in response to DNA damage. We exposed cells to ultraviolet light (UV-C) and imaged the nuclear distribution of PML, p53, and the BLM helicase by confocal microscopy. After DNA damage, PML partially relocated out of the PML-NBs, and colocalized with BLM and p53 at sites of DNA repair. In addition, using the isogenic HCT116 cell lines (p53+/+ and -/-), we show that the redistribution of PML was dependent on functional p53. Western analysis revealed that the level of PML protein remained unaltered after UV-C treatment. These results are consistent with the hypothesis that PML, in conjunction with p53 and BLM, contributes to the cellular response to UV-C-induced DNA damage and its repair. PMID- 12642866 TI - The oncoprotein MLL-ENL disturbs hematopoietic lineage determination and transforms a biphenotypic lymphoid/myeloid cell. AB - Mixed-lineage leukemia (MLL) fusion proteins are associated with a unique class of leukemia that is characterized by the simultaneous expression of lymphoid specific as well as myeloid-specific genes. Here we report the first experimental model of MLL. Murine bone marrow cells were retrovirally transduced to express the MLL-eleven nineteen leukemia (MLL-ENL) fusion protein. When cultivated in flt 3 ligand, stem cell factor and interleukin-7 (IL-7) in a stroma-free culture system MLL-ENL-transduced as well as control cells showed a wave of B lymphopoiesis. Whereas the controls exhausted their proliferative capacity in a CD19+/B220+ state, a continuously proliferating CD19-/B220+ cell population emerged in the MLL-ENL-transduced cultures. Despite the lymphoid surface marker, these cells were of monocytoid morphology. The immortalized cells contained unrearranged retrovirus, expressed MLL-ENL mRNA and were able to grow in syngenic recipients. From the diseased animals an MLL-ENL positive, B220+/CD19- cell type could be reisolated and cultivated in vitro. In analogy to human MLL, MLL-ENL transformed cells not only coexpressed lymphocyte-specific (rag1, rag2, pax5, Tdt) and monocyte-specific genes (lysozyme, c-fms), but also showed rearrangements of the genomic immunoglobulin locus. This model shows that MLL-ENL influences events of early lineage determination and it will enable the investigation of the underlying molecular processes. PMID- 12642867 TI - Naturally occurring dominant-negative Stat5 suppresses transcriptional activity of estrogen receptors and induces apoptosis in T47D breast cancer cells. AB - Signal transducer and activator of transcription (Stat) 5 regulates growth, differentiation, and survival of mammary and hematopoietic cells. The role of Stat5 in breast cancer has not been established, although Stat5 is critical for some hematopoietic malignancies. We detected for the first time that Stat5b is constitutively activated in human breast cancer cell lines, and analysed the role of Stat5 in estrogen receptor(ER)-positive breast cancer cell lines using dominant-negative variants of Stat5. Two distinct carboxyl-truncated Stat5a derivatives were generated. Stat5aDelta740 corresponded to a naturally occurring alternative splice variant, and Stat5aDelta713 was analogous to an 80 kDa Stat5a product of a nuclear protease. Stat5aDelta740 and Stat5aDelta713 displayed comparable dominant-negative properties and suppressed transcriptional activity of wild-type Stat5a and Stat5b equally well. Cotransfection experiments revealed that Stat5aDelta740 completely blocked transcriptional activity of endogenous estrogen receptor in T47D and MCF7 cells, and of both ER alpha and ER beta in COS 7 cells. Stat5aDelta740 was selected for adenoviral delivery, and high-efficiency expression of tyrosine phosphorylated Stat5aDelta740 was achieved in infected cells. Adenoviral-mediated Stat5aDelta740 induced apoptosis in T47D cells but not in caspase-3-negative MCF7 cells. The present study indicates that overexpression of a dominant-negative variant of Stat5 suppresses ER transcriptional activity and induces apoptosis in estrogen-responsive breast cancer tissue culture cells. PMID- 12642868 TI - Interferon-alpha sensitizes human hepatoma cells to TRAIL-induced apoptosis through DR5 upregulation and NF-kappa B inactivation. AB - Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), a member of the TNF superfamily, induces apoptosis in a variety of cancer cells with little or no effect on normal cells. Human hepatoma cells, however, are resistant to TRAIL-induced apoptosis. Since interferon-alpha (IFN-alpha) is capable of enhancing TNF-alpha-induced apoptosis in certain cancer cells, we evaluated the effect of IFN-alpha on TRAIL-induced apoptosis of human hepatoma cells. IFN-alpha pretreatment enhanced TRAIL-induced apoptosis of HuH-7 and Hep3B cells, in which IFN-alpha upregulated the expression of DR5, a death receptor of TRAIL, and downregulated the expression of survivin, which has an antiapoptotic function. In contrast, IFN-alpha did not enhance TRAIL-induced apoptosis of HepG2 cells, in which expression of DR5 and survivin was not affected by IFN-alpha. On the other hand, TRAIL activated NF-kappa B composed of RelA-p50 heterodimer, a key transcription factor regulating cell survival, in HuH-7 and HepG2 cells. However, IFN-alpha pretreatment repressed the TRAIL-mediated activation of NF-kappaB and decreased its transcriptional activity in HuH-7 but not in HepG2 cells. Moreover, IFN-alpha pretreatment clearly augmented TRAIL-mediated caspase-8 activation in HuH-7 cells. Our results suggest that IFN-alpha could sensitize certain human hepatoma cells to TRAIL-induced apoptosis by stimulating its death signaling and by repressing the survival function in these cells. PMID- 12642869 TI - Abnormal expression of 11 beta-hydroxysteroid dehydrogenase type 2 in human pituitary adenomas: a prereceptor determinant of pituitary cell proliferation. AB - The physiological effects of glucocorticoids (GCs) are, at least in part, mediated by inhibition of cell proliferation. Two isozymes of 11 beta hydroxysteroid dehydrogenase (11 beta-HSD) interconvert cortisol (F) and inactive cortisone (E), and are thus able to modulate GC action at an autocrine level. Previously, we have demonstrated absent expression of 11 beta-HSD2 in normal pituitaries; however, in a small number of pituitary tumors analysed, 11 beta HSD2 was readily demonstrable. Here we have used real-time RT-PCR to quantify expression of mRNA for 11 beta-HSD1 and 2 in 105 human pituitary tumors and have performed enzyme expression and activity studies in primary pituitary cultures. Overall, pituitary tumors expressed lower levels of 11 beta-HSDl mRNA compared with normals (0.2-fold, P<0.05). In contrast, expression of 11 beta-HSD2 mRNA was 9.8-fold greater in tumors than in normals (P<0.001). Enzyme assays showed significant 11 beta-HSD2 activity (71.9+/-22.3 pmol/h/mg protein (mean+/-s.d.)) but no detectable 11 beta-HSDl activity. Proliferation assays showed that addition of glycyrrhetinic acid (an 11 beta-HSD2 inhibitor) resulted in a 30.3+/ 7.7% inhibition of cell proliferation. In summary, we describe a switch in expression from 11 beta-HSDl to 11 beta-HSD2 in neoplastic pituitary tissue. We propose that abnormal expression of 11 beta-HSD2 acts as a proproliferative prereceptor determinant of pituitary cell growth, and may provide a novel target for future tumor therapy. PMID- 12642870 TI - Mutations of TP53 induce loss of DNA methylation and amplification of the TROP1 gene. AB - p53 and DNA methylation play key roles in the maintenance of genome stability. In this work, we demonstrate that the two mechanisms are linked and that p53 plays a role in the maintenance of the DNA methylation levels. The loss of p53 was shown to induce loss of DNA methylation in the TROP1 gene, a human cancer-expressed locus that undergoes amplification when hypomethylated. This demethylation was reverted by the reintroduction of a wild-type TP53 (wtTP53) in the TP53-null cells. Using a gene-amplification assay in vivo, we demonstrate that the loss of p53 leads to a demethylation-dependent TROP1 gene amplification. The induction of gene amplification was reverted by the expression of a wtTP53 gene or by in vitro methylation of the transfected DNA with the Sss I DNA methylase. Taken together, these findings demonstrate that the inactivation of TP53 induces loss of DNA methylation and DNA methylation-dependent gene amplification. PMID- 12642871 TI - Modulation of p53 and p73 levels by cyclin G: implication of a negative feedback regulation. AB - Cyclin G is a transcriptional target gene of tumor suppressor p53. Recent studies present evidence that cyclin G may play a central role in the p53-Mdm2 autoregulated module, but the precise function of cyclin G remains elusive. Here, we show a negative effect of cyclin G on the stability of p53 and p73. Cyclin G expression resulted in a dramatic decrease of p53 protein levels in response to DNA damage and abrogated irradiation-mediated G1 arrest along with an increase of S phase in MCF7 cells containing wild-type p53. In p53-null Saos2 cells, cyclin G inhibited p73 induction in response to genotoxic stress and delayed the camptothecin-mediated cell cycle arrest. Cyclin G interacts with p53 as well as p73, and its binding to p53 or p73 presumably mediates downregulation of p53 and p73. We also found that cyclin G-mediated reduction of p53 but not of p73 is Mdm2 dependent. Moreover, inhibition of cyclin G by small interfering RNA (siRNA) caused the accumulation of p53 and p73 protein levels in response to DNA damage. Therefore, our results imply that cyclin G is transcriptionally activated by p53 or p73, and, in turn, cyclin G negatively regulates the stabilization of p53 family proteins through an unknown mechanism different from ubiquitination or transcriptional control. PMID- 12642872 TI - alpha(v) integrins regulate cell proliferation through integrin-linked kinase (ILK) in ovarian cancer cells. AB - Integrins regulate both adhesion and signaling processes involved in proliferation and survival. alpha(v)beta(3) and alpha(v)beta(5) integrins have been shown to mediate cell adhesion and migration. Here we used human ovarian cancer cell lines (IGROV1, SKOV-3) that express alpha(v)beta(3) and alpha(v)beta(5) to study their role in cell proliferation and the signaling pathways involved. We found that alpha(v) integrins regulate cell proliferation through activation of integrin-linked kinase (ILK). An anti-alpha(v)-blocking antibody specifically inhibits the growth of IGROV1 and SKOV-3. The inhibition of cell proliferation involves alpha(v)beta(3) in IGROV1 cells, and both alpha(v)beta(3) and alpha(v)beta(5) in SKOV-3 cells. The reduced growth rate induced by alpha(v) integrin blockade is linked in both cell lines to G1/S cell cycle arrest. alpha(v) integrin blockade by neutralizing antibody as well as cyclic-RGD peptide caused an inhibition of ILK activity and phosphorylation of PKB/Akt on serine-473 but not on threonine-308, and was accompanied by an increase in p27(Kip1) expression. Overexpression of wild-type ILK rescued the phosphorylation of PKB/Akt on serine-473 in cells treated with anti-alpha(v) antibody. Inhibition of ILK by a pharmacological inhibitor results in inhibition of cell proliferation, PKB/Akt phosphorylation and increase of p27(Kip1). These results demonstrate that alpha(v) integrins regulate ovarian cancer cell proliferation through ILK. PMID- 12642873 TI - IGF-1 activates p21 to inhibit UV-induced cell death. AB - The insulin-like growth factor-1 (IGF-1) and its downstream effector Akt have been documented as survival factors in response to a variety of stress signals. In this study, we show that IGF-1 activates p21 protein expression in a p53 dependent manner. Inhibition of PI-3 kinase or ectopic expression of a dominant negative Akt blocks the effect of IGF-1 on the upregulation of p21 expression. In addition, IGF-1 prevents the UV irradiation-mediated suppression of p21 and MDM2 expression. Furthermore, p21 is important for IGF-1-mediated cell survival upon UV irradiation. Taken together, these data indicate that IGF-1 may activate p21 in executing its survival function upon genotoxic insults. PMID- 12642874 TI - Requirement of the coiled-coil domains of p92(c-Fes) for nuclear localization in myeloid cells upon induction of differentiation. AB - The nonreceptor tyrosine kinase Fes is implicated in myeloid cells differentiation. It has been observed that its localization can be cytoplasmic, perinuclear, or nuclear. To further characterize this point, we studied Fes subcellular localization in myeloid cell lines (HL60 and K562) and in COS1 cells. Fes was observed in both the nucleus and the cytoplasm of HL60, K562 cells overexpressing Fes and only in the cytoplasm of COS1 cells, suggesting that nuclear localization is cell context dependent. Moreover, in myeloid cells, the treatment with differentiation-inducing agents such as retinoic acid, phorbol esters and vitamin D, is followed by an increase of the oligomeric form of Fes in the nucleus. In fact, oligomerization seems to be necessary for translocation to occur, since Fes mutants missing the coiled-coil domains are not able to form oligomers and fail to localize in the nucleus. The active form of Fes is tyrosine phosphorylated; however, phosphorylation is not required for Fes to localize in the nucleus, since tyrosine kinase inhibitors do not block the translocation process. PMID- 12642875 TI - Loss of p27(Kip1) cooperates with cyclin E in T-cell lymphomagenesis. AB - Cyclin E and p27(Kip1) are key regulators for cyclin-dependent kinases (Cdks) acting at the G1-/S-phase transition of the cell cycle. Whereas cyclin E is required for the activation of Cdk2, p27(Kip1) is a specific Cdk inhibitor and can block cell division. High levels of cyclin E and low levels of p27(Kip1) expression have been associated with malignant lymphomas in humans; the level of p27(Kip1) is even considered of prognostic significance. However, mice that lack p27(Kip1) do not develop any malignant lymphomas despite a pronounced lymphoid hyperplasia in thymus and spleen. We have previously described transgenic mice that carry a construct in which the cyclin E cDNA is under the control of the CD2 promoter/enhancer region and thus express high levels of cyclin E in the T-cell compartment (CD2-cyclin E). These animals are not predisposed for T-cell lymphomas in the absence of other cooperating events. Here we show that T-cells from CD2-cyclin E mice that at the same time are deficient for p27(Kip1) show a significantly higher Cdk2 activity than cells from wild-type or single mutant animals. Accordingly, a higher percentage of T cells in S/G2/M phase is found in CD2-cyclin E/p27(Kip1-/-) mice. After a long latency period of over 200 days, these animals develop spontaneous monoclonal T cell lymphoma whereas none of the single CD2-cyclin E transgenic or the p27(Kip1)-deficient mice showed any sign of lymphoid malignancies. Our findings demonstrate that a deregulation of control mechanisms at the G1/S transition by the combination of high cyclin E levels in the absence of p27(Kip1) is sufficient to predispose mice to develop lymphoid malignancies and further support a role of p27(Kip1) as a tumor suppressor and of cyclin E as a dominant oncogene. PMID- 12642876 TI - Identification and characterization of CRG-L2, a new marker for liver tumor development. AB - Liver cancer is very common worldwide and the rates of hepatocellular carcinoma (HCC) have increased by over 70% in the last 2 decades in the US. Late diagnosis, because of the lack of clinical symptoms, and decreased hepatic function, because of underlying hepatic disease, lead to the extremely high mortality rates associated with HCC. Clearly, the identification of markers that are expressed early in the development of HCC and that are easily detected in high-risk patients would aid in early diagnosis and increased survival. We present the cloning and characterization of a novel gene, CRG-L2 (Cancer related gene-Liver 2), which displays high expression in murine and human hepatocellular carcinomas. Using in situ hybridization, we show that CRG-L2 mRNA levels are increased early during the development of liver tumors in C3H/HeJ mice, and that in normal tissues CRG-L2 mRNA is restricted to the murine testis and human placenta. Its restricted expression in normal tissues and unique early upregulation during tumor development make CRG-L2 an excellent candidate as a new clinical marker of HCC. PMID- 12642877 TI - Human squamous cell carcinomas lose a mortality gene from chromosome 6q14.3 to q15. AB - Normal human keratinocytes possess a finite replicative lifespan. Most advanced squamous cell carcinomas (SCCs), however, are immortal, a phenotype that is associated with p53 and INK4A dysfunction, high levels of telomerase and loss of heterozygosity (LOH) at several genetic loci, suggestive of the dysfunction of other mortality genes. We show here that human chromosome 6 specifically reduces the proliferation or viability of a human SCC line, BICR31, possessing LOH across the chromosome. This was determined by an 88% reduction in colony yield (P<0.001), following the reintroduction of an intact normal chromosome 6 by monochromosome transfer. Deletion analysis of immortal segregants using polymorphic markers revealed the loss of a 2.9 Mbp interval, centred on marker D6S1045 at 6q14.3-q15, in 6/19 segregants. Crucially, allelic losses of this region were not identified in control hybrids constructed between chromosome 6 and the BICR6 SCC cell line that is heterozygous for chromosome 6 and which showed no reduction in colony formation relative to the control chromosome transfers. This indicates that the minimally deleted region at D6S1045 is not the result of fragile sites, a recombination hot spot, or a feature of the monochromosome transfer technique. LOH of D6S1045 was found in 2/9 immortal SCC lines and was part of a minimally deleted region of line BICR19. Furthermore, allelic imbalance, consistent with LOH, was detected in 3/17 advanced SCCs of the tongue. These results suggest the existence of a suppressor of SCC immortality and tumour development at chromosome 6q14.3-q15, which is important to a subset of human SCCs. PMID- 12642882 TI - No comfort in the rural South: women living depressed. AB - Despite the widespread notion of the bucolic life in the country, major depressive disorder (MDD) is common among impoverished women in the rural South. Women with MDD seldom get treated because of the paucity of treatment available, the inability to pay for services because of no insurance, and the distance they must travel to reach care. Even if treatment was available, impoverished rural Southern women are unlikely to seek services because of cultural and social prohibitions. These include incongruence between the biomedical model of MDD and sociocultural explanations for its causes and manifestations, stigma, and traditional viewpoints of women that keep them isolated and invisible. Innovative treatment strategies must be devised for these women that are based on local views of MDD and its treatment, and people and monetary resources available in poor rural economies. Needed research with this population include ethnographic studies to gain understanding of the cultural factors associated with MDD and its treatment and evaluation of outreach, and other novel paradigms of rural service delivery including the use of nonprofessional personnel. Although the problems of treatment and research with this population are daunting, there is an opportunity for imagination, innovation, and creativity in devising local solutions to local problems. PMID- 12642883 TI - Metropolitan-nonmetropolitan differences in amount and type of mental health treatment. AB - This article examines the extent to which, conditional on receiving treatment, the type of care differs across metropolitan and nonmetropolitan areas. Using data from the Medical Expenditure Panel Survey (MEPS), the findings indicate that nonmetro residents who obtained mental health care (n = 2,381) have fewer mental health visits in a calendar year than their metro counterparts after adjusting for individual-level characteristics. Although observed rates of hospitalization and contact with physicians are higher in nonmetro areas than metro areas, this difference is attributable primarily to compositional differences between metro and nonmetro residents. PMID- 12642884 TI - Technology and rural mental health. AB - In addition to the specific and pervasive rural issues of isolation and suitability of services, the rural mental health system faces many of the same problems as the health system in general: access and increasing costs. The introduction of technology adds the unknown dimensions of acceptability and feasibility. Technology has the potential to decrease the gap in services and improve education, support, and connectedness between the client and the provider. As an alternative to traditional face-to-face contact for those in rural and geographically dispersed areas, the Internet potentially can bridge the disparities in health care access for rural mental health services. With an improved understanding based on research, demonstration studies of model applications, and evidence of outcomes, the emerging technologies can serve as tools to achieve the major goals of preventing, assessment, and treating serious mental illnesses in the rural communities with less barriers and stigma. PMID- 12642885 TI - Gender-related concerns of rural women with severe and persistent mental illnesses. AB - This study used focus groups to investigate the perceptions of rural women with severe and persistent mental illness (SPMI) about their gender-related health concerns. In a series of five focus groups conducted with 16 women at rural psychosocial clubhouses, women with SPMI discussed many gender-related issues, including unresolved grief over loss of children, isolation from family members, lack of sexual partners, diminished sexuality, and bodily changes secondary to medication side-effects. Women with mental illness felt like their roles as women was superseded by their role as mentally ill individuals. Rural women with SPMI have unmet needs for women's health. Psychiatric nurses have the optimal background for addressing gender-related concerns of rural women with SPMI. PMID- 12642886 TI - Vulnerability to alcohol and other drug disorders in rural women. AB - Alcohol and other drug disorders (AOD) are a growing problem among women, with estimates as high as 11 million U.S. women affected. This article is a report of the results from 2 studies investigating risk factors for AOD disorders in rural women. Samples of the 2 studies were combined for a total of 267 women. Analyses focused on victimization as a vulnerability factor for AOD disorders in rural women. Comorbid Axis I mental disorders and nursing implications are discussed. PMID- 12642888 TI - Residential and nursing home care--issues of balance and quality of care. PMID- 12642887 TI - Shortages of rural mental health professionals. AB - This study presents a conceptual model of the supply and demand for mental health professionals. It uses national data to profile differences in the supply of mental health professionals in different types of rural and urban areas. It contrasts the availability of general health and mental health professionals. It examines shortage areas identified in 2000 and their related community characteristics. Because of the absence of data on a national level to describe many types of mental health professionals state licensure data for one state were used to show the volume and distribution of these practitioners. To improve rural mental health service delivery it will be necessary to implement system changes to promote the increased availability, competency, and support of rural health professionals. PMID- 12642889 TI - Does assessment make a difference for people with dementia? The effectiveness of the Aged Care Assessment Teams in Australia. AB - The needs of individuals with dementia and other psychiatric problems of old age have received increased attention in Australia over the last decade. This paper reports on the role of Aged Care Assessment Teams (ACATs) in managing these clients, and the extent to which they are differentiated from other clients in the assessment process and outcomes recommended. Data on some 26,500 clients seen by ACATs in Victoria in the second half of 1999 are analysed to show (1) the relationship between a diagnosis of dementia and reporting of disability in orientation, (2) characteristics of clients with and without a diagnosis of dementia and (3) outcomes for groups of clients defined on the basis of a diagnosis of dementia and disability in orientation. PMID- 12642890 TI - Public funding for residential and nursing home care: projection of the potential impact of proposals to change the residential allowance in services for older people. AB - BACKGROUND: This paper investigates the potential effects of a policy change in the funding of UK residential care. The White Paper Modernising Social Services (Cm 4169, 1998) outlined plans to change the distribution of the Residential Allowance (RA), payable in support of residents in independent residential or nursing home care, from a component of income support paid direct to establishments to a grant to local authorities. This change was intended to remove the perverse incentive in accessing independent residential care more favourably than local authority care. A further objective was to encourage local authorities to use the grant to support home-based alternatives to residential care. The policy rests on a model in which price signals dictate the choice of care for an older person. By, in effect, raising the price of independent residential and nursing home care, the policy provides an incentive for authorities to seek alternatives to institutional care. METHODS: Managers from 16 UK social services departments attended a focus group discussion, completed questionnaires and provided information to assist in calculating the potential diversionary effect of the policy. RESULTS: Managerial estimates indicated a small diversionary effect of the policy; A potential effect of 0.26 and 0.19 per 1000 older people diverted from residential and nursing care respectively. CONCLUSIONS: The study indicated that wider organisational factors other than price are likely to play a greater role in deciding whether an older person is admitted to care. Changes in public funding alone do not reflect the complexities involved in decision-making concerning the residential placement of older people. PMID- 12642891 TI - The long term care insurance law in Japan: impact on institutional care facilities. AB - BACKGROUND: Following the introduction of the public long-term care (LTC) insurance in Japan on 1 April 2000, funding that had been split between the health and social welfare sectors was unified. All elderly people 65 and over have become entitled to receive benefits according to their eligibility level, regardless of income or family support. Except for those assessed as in the least dependent group, individuals can choose either community care or institutional care, with only the cost of food an additional payment for the latter. METHOD: Review of the development and implications of long term care insurance provision for older people and analysis of quality based on resident level data. RESULTS: Despite the structural changes, the impact of the new programme has had minimal impact upon the providers of institutional care. The three types of facilities that had provided LTC have continued to function independently, basically retaining the same staffing requirements and charges. For indicators adjusted for risk, the quality of care tended to be higher in designated LTC hospital beds. CONCLUSIONS: Appropriate balance between institutional and community care and triaging among the three different types of facilities remains difficult because decisions rest with the individual. Indicators of quality must take into account differences in case-mix among the facility types. However, there are few incentives for providers to improve quality because demand is likely to be increasingly greater than supply. Whether the generous provisions of the new programme will prove to be sustainable remains to be seen. PMID- 12642892 TI - International experiments in integrated care for the elderly: a synthesis of the evidence. AB - BACKGROUND: The OECD countries have recently promoted policies of deinstitutionalisation and community-based care for the elderly. These policies respond to common cost pressures associated with population aging, and the challenge of providing improved care for the elderly. They aim to substitute less costly services for institutional ones, to improve patient satisfaction and decrease expenses. However, views concerning their success are mixed. We took a comparative cross-national approach to examine the evidence, to identify common features of an effective system of integrated care, and to examine the potential of such models to positively affect care of the elderly, and public finances. METHODS: We conducted a systematic review of recent demonstration projects testing innovative models of care for the elderly in OECD countries. Projects included aimed to create comprehensive integration of acute and long-term care services, and were evaluated using a comparison group. RESULTS: For each project, we report available results on rates of hospitalisation, long term care institutionalisation, utilisation and costs, impact on process of care, and health outcomes. In addition, the following common features of an effective integrated system of care were identified: a single entry point system; case management, geriatric assessment and a multidisciplinary team; and use of financial incentives to promote downward substitution. CONCLUSIONS: Community based care can impact favourably on rates of institutionalisation and costs. Comprehensive approaches to program restructuring are necessary, as cost effectiveness depends on characteristics of the system of care. Expansion of successful programmes to achieve widespread use remains a critical challenge. PMID- 12642893 TI - The cost implications of the changing population and characteristics of care homes. AB - BACKGROUND: Recent increases in care home closures suggest that homes may not be able to balance pressures to reduce costs against pressures to increase standards. Commissioning requires an understanding of the factors affecting costs and how they change over time. METHODS: A survey of care homes for older people was conducted in 21 local authorities in England in 1996. A complete response was obtained for 618 homes (75%) and 11,900 residents. Findings were compared with surveys conducted in 1986 and 1988. RESULTS: Dependency was significantly related to prices, primarily due to the differential payments to nursing and residential homes. Home characteristics were also related to price, the proportion of single rooms having the largest impact. However, prices were most sensitive to local wage rates, particularly in residential homes. Compared with previous surveys levels of dependency had increased, particularly in voluntary residential homes and nursing homes. Independent homes were more likely to be purpose built, and a higher proportion of beds were in single rooms, although only 30% of private residential, dual registered and nursing homes achieved the proposed level of 80% of beds in single rooms. Staffing ratios appeared to have increased, but price rises were modest, particularly for nursing homes. CONCLUSIONS: Standards of provision have improved over time, although prices appear to have been kept below those expected from increases in costs. Continuing pressures on costs and prices are likely to lead to further closures and a restriction of choice for older people. PMID- 12642894 TI - The role of the specialist physician in nursing homes: the Netherlands' experience. AB - BACKGROUND: Dutch nursing homes employ physicians who have completed a two year specialist training programme to become a qualified nursing home physician. This paper describes the origin of the profession of nursing home medicine in the context of Dutch nursing homes. METHOD: Review of the history and development of the need for specialist physicians in care of residents in nursing homes. Examination of the competencies required and training available for physicians and the benefits of this arrangement in improving quality of care. RESULTS: Relevant skills and competencies of the specialist physician in nursing homes are discussed. The role and tasks, and the nursing home physician's method of working and providing care are explained, as well as the benefits and disadvantages of organizing medical care in this way. CONCLUSIONS: The benefits of a specialist physician are considered greater than the disadvantages. Specialist training for this role is considered to be highly desirable. PMID- 12642895 TI - Improving nursing home quality of care through outcomes data: the MDS quality indicators. AB - BACKGROUND: The quality of care provided to nursing home residents is a continuing source of concern throughout the world. In the United States, the Health Care Financing Administration mandated the use of a standardised resident assessment instrument, called the Minimum Data Set (MDS) which facilitated the development of a set of nursing home quality of care indicators. METHODS: The MDS Quality Indicators were developed by a team of researchers at the University of Wisconsin-Madison and covered 12 domains, as well as associated risk factors. This initial set of indicators then went through an iterative process of review by national clinical panels, further empirical analysis, validation studies, and pilot tests, to confirm whether they were accurate, reliable and useful tools for identifying quality problems. RESULTS: The final set of MDS Quality Indicators includes 24 variables that include both processes and outcomes of care and prevalence and incidence measures. They are defined at both the resident and facility level. Four of the indicators are risk-adjusted. CONCLUSION: The indicators have several possible applications: a source of consumer information; to help guide policy makers; and to assist providers in conducting quality improvement initiatives and to evaluate the impact of these efforts. PMID- 12642896 TI - Benchmarking and quality in residential and nursing homes: lessons from the US. AB - BACKGROUND: Performance measurement and benchmarking are common concerns in the delivery of long term care. It is common to measure the performance of providers and to publicly report these data. This paper examines selected technical challenges facing those who design, implement and disseminate health care quality performance measures. METHOD: Review of the application of measures of performance in the US nursing home sector. RESULTS: Using examples drawn from the skilled nursing home arena, problems ranging from data reliability and validity, the multi-dimensional nature of quality measures and selection bias as well as differential measurement abilities are discussed. CONCLUSIONS: Benchmarking of performance is an inherently complex issue. However, to ensure that such comparisons are both fair and valid requires measures to be more technically sophisticated and sensitive to real changes attributable to changes in care. PMID- 12642897 TI - Conjugation and deconjugation of ubiquitin regulating the destiny of proteins. AB - The homeostasis for a number of cellular proteins is regulated by not only phosphorylation and dephosphorylation, but also ubiquitination and deubiquitination. A number of proteins involved in the degradation of polypeptides have been isolated in various eukaryotic organisms from Saccharomyces cerevisiae to human. Recently, several deubiquitinating enzymes, classified into either the Ub C-terminal hydrolase (UCH) or the Ub-specific processing protease (UBP), have been reported. It has been shown that they contain conserved domains including Cys, His, and Asp residues throughout the enzyme. These proteins have been demonstrated that Cys and His domains are critical for deubiquitinating enzymatic activity. Recently, we have shown that the Asp domain localized between Cys and His domains is also essential for cleaving the ubiquitin from protein substrates. Mouse deubiquitinating enzymes including DUB-1, DUB-2, and DUB-2A have been isolated and they showed the expression specificity. Of these, DUB- 1 and DUB-2 are expressed in lymphocytes depending on the presence of cytokines (interleukin-3 in B-lymphocytes and interleukin-2 in T- lymphocytes, respectively), indicating that they are involved in cytokine signaling pathways. Isolation of all putative DUBs will help to identify their substrates and to regulate the homeostasis of cellular proteins, especially in proliferative cells. PMID- 12642898 TI - Involvement of tumor necrosis factor receptor superfamily(TNFRSF) members in the pathogenesis of inflammatory diseases. AB - Current therapies for autoimmune diseases are not cures but merely palliatives, aimed at reducing symptoms. For the most part, these treatments provide nonspecific suppression of the immune system and thus do not distinguish between a pathogenic autoimmune response and a protective immune response. Recently emerging evidence not only has indicated the involvement of members of the TNF receptor/ligand superfamilies but also has revealed exciting innovative strategies for the treatment of autoimmune diseases and other chronic inflammatory diseases without depressing the immune response in general. In this review, we will discuss the regulatory mechanisms of TNF receptor/ligand family members, such as HVEM/ LIGHT, 4-1BB/4-1BBL, and GITR/GITRL that regulate T and B cell functions and participate in the process of inflammatory diseases. We will also discuss how intervening in the costimulatory pathways mediated by these molecules might have some potential as a therapeutic approach to immune disorders. PMID- 12642899 TI - Enzymatic method for assaying calcium in serum with Ca++ -ATPase. AB - A kinetic assay for total calcium in serum was developed which is based on the activation of Ca(++)-ATPase by free Ca(++) [Ca(++)](f) maintained by EGTA in the reaction mixture. The concentration of Ca(++)(f) was dependent on total reference calcium added or serum calcium. Ca(++)-ATPase activity was coupled to the reduction of NADH by pyruvate kinase (PK) and lactate dehydrogenase (LDH) and monitored by change in absorbance at 340 nm. The calcium in normal serum was 10.08 +/- 0.24 mg/dl (n = 35) by our method while with o-cresolphthalein complexone (CPC) method, the total calcium in the same 35 serum samples was 10.14 +/- 0.54 mg/dl. The range of within-run coefficient of variations (CVs) by this method was 0.9-2.87% at 8-12 mg/dl and day-to-day CVs were 0.72-3.17%. The presence of other ions and standard clinical interfering agents did not affect this assay system. The correlation between values obtained with our method (y) and CPC method (x) for normal serum was: y = 1.064x-0.580 mg/dl (r = 0.912, n = 59). PMID- 12642900 TI - Acetyl-CoA carboxylase beta expression mediated by MyoD and muscle regulatory factor 4 is differentially affected by retinoic acid receptor and retinoid X receptor. AB - Mammals have two major isoforms of acetyl-CoA carboxyase (ACC). The 275 kDa beta form (ACCbeta) is predominantly in heart and skeletal muscle while the 265 kDa alpha-form (ACCalpha) is the major isoform in lipogenic tissues such as liver and adipose tissue. ACCbeta is thought to control fatty acid oxidation by means of the ability of malonyl-CoA to inhibit carnitine palmitoyl-CoA transferase-1 (CPT 1), which is a rate-limiting enzyme of fatty acid oxidation in mitochondria. Previously, it was reported that MyoD and other muscle regulating factors (MRFs) up-regulate the expression of ACCbeta by interactions between these factors and several cis-elements of ACCbeta promoter. We described here that ACCbeta expression mediated by MRFs is regulated by retinoic acids. Endogenous expression of ACCbeta in differentiated H9C2 myotube was significantly increased by retinoic acid treatment. However, on transient transfection assay in H9C2 myoblast, ACCbeta promoter activity was suppressed by RXRalpha and more severely by RARalpha. These effects on ACCbeta expression in myoblasts and myotubes by RXRalpha and RARalpha seem to be mediated by their interactions with MRFs because no consensus sequence for RXRalpha and RARalpha has been found in ACCbeta promoter and retinoic acid receptors did not affect this promoter activities by itself. In transient transfection in NIH3T3 fibroblast, the activation of ACCbeta promoter by MyoD, main MRF in myoblast, was significantly suppressed by RARalpha and to a less extent by RXRalpha while the RXRalpha drastically augmented the activation by MRF4, major MRF in myotube. These results explained that retinoic acids differentially affected the action of MRFs according to their types and RXRalpha specially elevates the expression of muscle specific genes by stimulating the action of MRF4. PMID- 12642901 TI - Metastasis-suppressor KAI1/CD82 induces homotypic aggregation of human prostate cancer cells through Src-dependent pathway. AB - To investigate the functional role of KAI1/CD82, a metastasis suppressor for human prostate cancer, in the regulation of homotypic cell adhesion, we transfected KAI1 cDNA into DU 145 human prostate cancer cells and established stable transfectant clones with high KAI1/CD82 expression. The KAI1 transfectant cells exhibited significantly increased homotypic cell aggregation in comparison with the control transfectant cells. This aggregation of the KAI1 transfectants was further enhanced upon exposure to anti-CD82 antibody, suggesting that KAI1/CD82 may be involved in the intracellular signaling for the cell adhesion. Among several signal pathway inhibitors tested, PP1, an inhibitor of Src family kinases, significantly suppressed homotypic aggregation of the KAI1 transfectant cells. Ligation of KAI1/CD82 with anti-CD82 antibody increased endogenous Src kinase activity of the KAI1 transfectant cells. When different types of src expression constructs were retransfected into the KAI1-transfected DU 145 cells, kinase-negative mutant src transfectant cells exhibited much lower homotypic aggregation than the mock cells transfected with an empty vector. Moreover, homotypic aggregation of the mutant src transfectant cells was not enhanced by KAI1/CD82 ligation with anti- CD82 antibody. These results suggest that Src mediates the intracellular signaling pathway of KAI1/CD82 for the induction of homotypic adhesion of human prostate cancer cells. PMID- 12642902 TI - Anti-apoptotic role of phospholipase D isozymes in the glutamate-induced cell death. AB - Phospholipase D (PLD) plays an important role as an effector in a variety of physiological processes that reveal it to be a member of the signal transducing phospholipases. Recently, PLD2 was reported as a necessary intermediate in preventing apoptosis induced by hydrogen peroxide or hypoxia in rat pheochromocytoma (PC12) cells. The data presented here show that both PLD isozymes, PLD1 and PLD2 are also required in attenuating glutamate-induced cell death in PC12 cells. Treatment of PC12 cells with glutamate resulted in induction of apoptosis in these cells, which is accompanied by decreased PLD activity and increased ceramide concentration. Incubation of PC12 cells with exogenous C6 ceramide showed a time-dependent decrease of PLD activity. When cDNAs of PLD1 and PLD2 were transfected into PC12 cells respectively, overexpression of PLD1 or PLD2 resulted in inhibition of glutamate-induced apoptotic cell death. These data indicate that both PLD1 and PLD2 play a protective role against glutamate-induced cell death in PC12 cells. PMID- 12642903 TI - Five subtypes of muscarinic receptors are expressed in gastric smooth muscles of guinea pig. AB - Muscarinic receptors play key roles in the control of gastrointestinal smooth muscle activity. However, specific physiological functions of each subtype remain to be determined. In this study, the nonselective cation channel activated by carbachol (I(CCh)) was examined in circular smooth muscle cells of the guinea pig gastric antrum using patch-clamp technique. 4-DAMP inhibited I(CCh) dose dependently with IC(50) of 1.1 0.1 nM (n = 6). GTPgS-induced current, however, was not inhibited by 10 nM 4-DAMP. I(CCh) was not recorded in pertussis-toxin (PTX)-pretreated smooth muscle cells of gastric antrum. I(CCh) values in response to 10 mM CCh at a holding potential of 60 mV were -330 32 pA (n=4) and -15 3 pA (n = 6) in the control and PTX-treated cells, respectively (P 0.01). Sensitivities to nanomolar 4-DAMP and PTX suggest the possible involvement of m4 subtype. Using sequence information obtained from cloned guinea pig muscarinic receptor genes, it is possible to amplify the cDNAs encoding m1-m5 from guinea pig brain tissue. Single cell RT-PCR experiments showed that all five subtypes of muscarinic receptor were present in circular smooth muscle cells of the guinea pig gastric antrum. Together with our previous results showing that G(o) protein is important for activation of ACh-activated NSC channels, our results suggest that I(CCh) might be activated by acetylcholine through m4 subtype as well as m2 and m3 subtypes in guinea-pig stomach. PMID- 12642905 TI - [Roles of brain science in psychiatry]. AB - At the opportunity of celebrating the 100th Anniversary of the Japanese Society of Psychiatry and Neurology, I am filled with deep emotion when I recall the time only a few decades ago, when mental disorders were believed to be an illness of the mind without any disorder in the brain. It has now become apparent that a mental disease emerges from the malfunction of the brain's unique biological mechanisms, so that any mental disease can be cured or prevented if the cause of the malfunction is clarified. Analyses of intercellular as well as intracellular signal transduction and the mechanisms underlying gene regulation in brain cells have recently advanced markedly and have brought a marked improvement in the methods and technology for investigating pathogenesis of neurological and psychiatric diseases. These impressive results have been derived in the search for the cause of Parkinson's disease, Alzheimer's disease and spinocerebellar degeneration. Certain minor changes in the molecular structure of sodium channels have been found to cause epilepsy. It has also been revealed that ten particular sites in chromosomes harbor the remote cause of schizophrenia and depression. These sites are probably responsible for production of certain synaptic transmitters and modulators. Another major challenge in the field of psychiatry is to understand the unique symptoms exhibited by patients with psychiatric disorders, which requires research to head in a direction different from molecular and cellular brain science. It is a challenge similar to the very fundamental challenge of determining how our brain, which is composed of numerous brain cells, yields our mind (or, should I say, supports our mind). To understand the brain as a complex system is more difficult than researching its molecular and cellular machinery, and accordingly, the progress of research in this field has been slow. However, it is gradually gaining speed thanks to the improvement in non-invasive methods for measuring activities in conscious human brains as well as that to the establishment of computational models. For example, it is a major mystery why a schizophrenic patient experiences auditory hallucinations, but an interesting hypothesis is derived by considering activities of the thinking brain as a neural system. Under this hypothesis, we gather information from the outside world by using our sensory systems and establish its internal model in our brain. Thus, manipulation of the internal model to run a simulation is an important part of our thinking activities. When an internal model behaves abnormally, abnormal information is generated, which makes the patient mistake it for sound from the outside, thus causing auditory hallucinations. It is highly expected that substantial progress will be made in psychiatry through proposals of new hypothetical models such as this one as well as verification of them through non-invasive measurement of brain activities. In the near future, patients with mental disorders will most likely benefit from the results of this two-directional research in psychiatry. The expectation is high for the strong leadership of the Japanese Society of Psychiatry and Neurology to promote research in Japan. Effective research effort requires efficient collection of human materials and systematic survey of patients, which will inevitably increase the involvement of patients in basic research. Improvement of research environments such as by developing a method for assessing research ethics and upgrading brain banks is urgently needed. PMID- 12642904 TI - Soluble factor from tumor cells induces heme oxygenase-1 by a nitric oxide independent mechanism in murine peritoneal macrophages. AB - Tumor target-derived soluble secretary factor has been known to influence macrophage activation to induce nitric oxide (NO) production. Since heme oxygenase-1 (HO-1) is induced by a variety of conditions associated with oxidative stress, we questioned whether soluble factor from tumor cells induces HO-1 through NO-dependent mechanism in macrophages. We designated this factor as a tumor-derived macrophage-activating factor (TMAF), because of its ability to activate macrophages to induce iNOS. Although TMAF alone showed modest activity, TMAF in combination with IFN-gamma significantly induced iNOS expression and NO synthesis. Simultaneously, TMAF induced HO-1 and this induction was slightly augmented by IFN-gamma. Surprisingly, however, induction of HO-1 by TMAF was not inhibited by the treatment with the highly selective iNOS inhibitor, 1400 W, indicating that TMAF induces the HO-1 enzyme by a NO-independent mechanism. While rIFN-gamma alone induced iNOS, it had no effect on HO-1 induction by itself. Collectively, the current study reveals that soluble factor from tumor target cells induces HO-1 enzyme in macrophages. However, overall biological significance of this phenomenon remains to be determined. PMID- 12642906 TI - [Are psychological concepts of Japanese origin relevant?]. AB - This paper was originally given as a lecture in English at the XII World Congress of Psychiatry held in Yokohama, August 2002. Its primary aim was to introduce Morita therapy and the amae psychology to participants from abroad, not as something exotic, but as something intrinsically relevant to them. In my opinion, Morita therapy is based upon Morita's discovery that certain neurotics, whom he eventually named as shinkeishitsu, can recover spontaneously from severe anxiety when placed in isolation and forced to rest in bed without any diversion under the doctor's strict supervision. In other words, only those patients who follow the doctor's advice faithfully through initial isolation and subsequent work stages can benefit from Morita therapy. It is further argued that it does not do justice to Morita to speculate that he developed his theory and therapy under the influence of Zen. Apropos of the amae psychology what is most noteworthy about it is that its genuine feeling can be conveyed only nonverbally unlike love which one can express verbally when one says "I love you". It is understood that ambivalence refers to the co-existence of love and hate. This love, however, had better be called amae since it is usually nonverbal and not even acknowledged. Narcissism also entails amae, in fact amae turned upon itself, because narcissism consists in denying one's virtual dependence. Interestingly, the word amae alone may suggest a narcissistic state when it is used in the sense of self-indulgence, the usage which is now becoming quite common. Apart from elucidating two psychiatric terms, the concept of amae turns out to be quite handy in describing what transpires in the psychotherapeutic situation including that of Morita therapy. The concluding remark is to emphasize the importance of psychological concepts against the prevailing view of evidence-based psychiatry. It is argued that the objectivity of scientific findings is in the final analysis a postulate guaranteed by the conscience of individual scientists. If so, the same kind of objectivity may be attributed to the observations made by the doctor when he maintains the psychotherapeutic relationship against all odds and also without any compromise of his conscience. PMID- 12642907 TI - Empathy, meaning, and the doctor-patient relationship. PMID- 12642908 TI - Prevention and early intervention in schizophrenia: facts and visions. AB - There are large-scale preventive programmes to reduce the risk of death and disability caused by several frequent physical diseases. The primary and secondary prevention of schizophrenia, a disorder entailing many years of life in disability, is still being neglected. Prevention is aimed at reducing the incidence, severity or consequences of the disorder. To find ways of preventive intervention in schizophrenia, the aetiological risk factors must be identified, then eliminated or modified. As possible targets pre-, peri- and postnatal complications, urbanicity and early behavioural risk indicators are discussed. As examples of successful early prevention targeted at risk indicators attempts to prevent depression and violence are considered. The most promising approach at present is secondary prevention focused on early illness course. Based on a controlled retrospective assessment of 232 first illness episodes the course of prodromi, impairments and psychotic symptoms prior to the climax of the first episode is shown. Most of the social consequences occur before the first treatment contact, thus making plain the urgent need for preventive action. Tools sufficiently validated are not yet available for early diagnosis and prediction of psychosis onset at the prepsychotic stage. So intervention has to be based on high-risk inclusion criteria, which exclude large proportions of at-risk persons. Appropriate early intervention at the prepsychotic, prodromal and the early psychotic stage as well as relevant ethical considerations are discussed. The frequency of and distress associated with single psychotic symptoms in the general population are potent predictors of a psychosis. The vision of treating this early illness dimension with third-generation, side-effect-free antipsychotics or of preventing its onset by oestrogen-like substances is discussed. PMID- 12642909 TI - [Molecular biology of drug dependence and behavioral sensitization]. AB - Molecular mechanisms in the development of drug abuse and dependence were reviewed by taking behavioral sensitization induced by psychostimulants like amphetamines and cocaine as a typical example. Behavioral sensitization is characterized by three main features, progressive quantitative and qualitative changes in responsiveness to the drug, very long-lastingness, and development of vulnerability to other drugs and nonspecific physical and psychological stressors, in other words, cross-sensitization. These serial changes in response to the drug during abuse must result from plastic changes in the brains of abusers. As to subcellular neurochemical mechanisms of sensitization, the activation of three main cascades is indispensable, 1) D1 dopamine (DA) receptors/PKA/phospho-34Thr-DARPP-32/PP-1 cascade activated by psychostimulant induced enhancement of DA release in the accumbens, 2) NMDA receptors and CaM-KII activated by enhanced release of glutamate, 3) activation of MAP kinase cascade by BDNF and beta 1 subunit of G protein. These, in turn, activate several transcription factors, including delta-Fos B, and affect transcription and translation of 4th or later messengers. Finally, these result in the rearrangement of neural networks, where the tone of the A10 dopamine pathway from the ventral tegmentum area to the accumbens is strengthened, and regulation by glutamatergic afferents from the frontal cortex, amygdala and hippocampus shifts into abnormal positive regulation. As amphetamines increase expression of some plasticity-related genes (e.g. synaptophysin, stathmin and arc), synaptogenesis, neuritic sprouting and elongation must develop during behavioral sensitization. These plastic changes with structural modification of neural networks in the CNS during drug abuse could induce and reinforce psychological dependence and susceptibility to drug-induced psychoses, which become increasingly intractable. PMID- 12642910 TI - [Classification of long-term clinical course of 'atypical psychosis': a 20-year follow-up study at a medical school hospital]. AB - To study the long-term clinical course and outcomes of atypical psychosis, 8 patients diagnosed with atypical psychosis were observed for more than 12 years (mean, 20 years). Retrospective examination was performed, particularly with respect to clinical features at each episode. The overall course of each case was classified as one of the following three types: Type I--"Recurrent confused state" type. Patients frequently repeated acute transient confused or dream-like states in a similar way, sometimes and/or for part of the episode accompanied by a floating paranoid-hallucinatory state. Duration of psychotic episode was short, persisting for a few days to about one month. Type II--"Manic-depressive illness similar" type. After a long course of disease, the predominantly early middle aged patients (30- to 40 years-old) demonstrated fewer original characteristic features of acute confused or dream-like states. Instead, manic or depressive episodes tended to predominate. Duration of psychotic episodes exceeded the duration of type I episodes, to a maximum of about 3 months. Type III- "Appearance of residual state" type. After several episodes characterized by transient confused state during middle age, residual states consisting of a slight depressive state, reduced spontaneity and flattening of emotions appear. These states become durable and the periodicity of the disease disappeared. We conclude that the core group of atypical psychosis patients presents with confused symptoms as a clinical feature of episodes, and with the recurrent confused state type representing the long-term clinical course. "Shift to manic depressive illness similar" and "appearance of residual state" types were considered to be derived from the core group, according to the interplay of personality structure and viable dynamics. PMID- 12642911 TI - [Effects of continuous positive airway pressure on Cheyne-Stokes breathing in congestive heart failure]. AB - To investigate the effects of a continuous positive airway pressure (CPAP) of 5 cmH2O for three months on sleep disordered breathing and cardiac function in congestive heart failure (CHF), we performed physiological tests before and after the CPAP treatment in six male patients with stable CHF, whose echocardiographic left ventricular ejection fraction (LVEF) was below 30%. The control polysomnography revealed that all six had Cheyne-Stokes breathing (with central sleep apnea in four, and central sleep hypopnea in two). Prior to the CPAP, the number of episodes of apnea/hypopnea per hour of sleep, the contribution of slow wave sleep to total sleep time, LVEF and the cardio-thoracic ratio were 34.7 +/- 21.4, 4.1 +/- 1.5%, 24.4 +/- 2.3% and 58.4 +/- 3.7% (mean +/- SD), respectively. After the CPAP, these values changed to 6.0 +/- 7.0, 16.5 +/- 8.3%, 34.3 +/- 3.3% and 55.0 +/- 4.2%, respectively, all of which changes showed a statistical significance (p < 0.05). We conclude that CPAP is an effective treatment for CHF with Cheyne-Stokes breathing, improving sleep disordered breathing, sleep quality and cardiac function, and providing a new non-pharmacological approach for CHF. PMID- 12642912 TI - [Clinicopathological analysis of patients with idiopathic pulmonary fibrosis which became acutely exacerbated after video-assisted thoracoscopic surgical lung biopsy]. AB - Nine patients undergoing video-assisted thoracoscopic surgical (VATS) lung biopsy over a five-year period from 1997 to 2001 with the ultimate diagnosis of usual interstitial pneumonia without underlying connective tissue disease were identified. In two of nine patients, acute exacerbation occurred six days after VATS lung biopsy. We reviewed the clinical records and pathology of all nine cases, and found that the two cases of exacerbation had higher peripheral white blood cell counts and KL-6, lower PaO2, VC and FEV 1, and a longer inhalation of FIO 2 = 1.0 during VATS, and needed a longer period of chest drainage after VATS. Abundant inflammatory cell infiltration and fibroblastic foci were observed in the exacerbation cases. Thus, patients with usual interstitial pneumonia of the idiopathic type, who have high disease activity and low pulmonary function, may be at high risk of acute exacerbation following VATS lung surgery. PMID- 12642913 TI - [A case in which progressive emphysematous changes were seen on chest computed tomography associated with Pneumocystis carinii pneumonia in a patient with AIDS]. AB - A 60-year-old man was admitted to our hospital complaining of progressive dyspnea. On admission, he had marked hypoxemia, and his chest radiography and computed tomography (CT) showed ground glass opacities and multiple emphysematous changes in both lung fields. On examining the patient's bronchoalveolar lavage fluid (BALF). Pneumocystis carinii pneumonia (PCP) was diagnosed. A serological test for human immunodeficiency virus (HIV)-1, 2 was positive, and acquired immunodeficiency syndrome (AIDS) was diagnosed. Since the chest CT performed a month before the patient's admission to our hospital revealed ground glass opacities in both lung fields we thought that he had already developed PCP at that time. In comparison with his previous CT, the chest CT on admission showed progressive ground glass opacities and emphysematous changes. Although PCP is known to display various findings on chest radiography and CT, emphysematous changes are rarely reported in Japan. In this case we were able to confirm these changes and observe its progression using chest CT. PMID- 12642914 TI - [A case of neurogenic pulmonary edema associated with epileptic seizure]. AB - We report a case of neurogenic pulmonary edema associated with epileptic seizure. A 36-year-old woman had had several episodes of fainting and postictal respiratory failure, and since July 1998 had been admitted to a nearby hospital three times. On October 12, 1999, she was again admitted to a nearby hospital with the same symptom, and was transferred from that hospital to ours for evaluation of the recurrent respiratory disorder. Low-grade fever, mild leukocytosis, hypoxemia and bilateral diffuse opacities were observed as previously on chest radiography, and improved within several days without any specific therapy. The negative C reactive protein level, normal cardiac function and faintly bloody bronchoalveolar lavage fluid were also observed. There was no evidence of aspiration pneumonia, infectious disease, or underlying heart or lung disease. Electroencephalography showed spikes in accord with the left temporal lobe, and the cause of the patient's fainting was thought to be temporal lobe epilepsy. After all other causes had been excluded, this case was diagnosed as neurogenic pulmonary edema associated with epileptic seizure. Only about 40 cases of the postictal pulmonary edema have been reported since 1908, and the pathophysiologic mechanism of this condition is still unknown. Neurogenic pulmonary edema associated with epileptic seizure is rare, but the importance of awareness of this condition needs to be emphasized because it is suspected to be the cause of unexpected sudden death in epileptics. We should consider the disease as important in the differential diagnosis of acute respiratory failure associated with epilepsy. PMID- 12642915 TI - [A HTLV-I carrier initially treated for Pneumocystis carinii pneumonia]. AB - A 65-year-old woman was admitted to the hospital with complaints of dyspnea on effort Chest radiography and CT revealed bilateral interstitial infiltration. Transbronchial lung biopsy specimens revealed organisms of Pneumocystis carinii. Sulfamethoxazole-trimethoprim improved the patient's radiograph and CT, and although a positive reaction was obtained for anti-HTLV-I antibody, the hematological findings and clinical symptoms did not suggest that she had an ATL. Since discharge, she has been receiving maintenance therapy at the outpatient clinic because an opportunistic infection is predictive of the development of ATL. PMID- 12642916 TI - [Bronchioloalveolar carcinoma complicated by a lesion resembling bronchiolitis obliterans organizing pneumonia in the opposite lung]. AB - A 50-year-old woman with abnormal shadows in her chest radiograph was admitted for a more detailed examination in April 2000. A chest CT scan showed infiltrates surrounded by ground-glass opacities in the right upper lobe, and ground-glass opacities with partial infiltrative changes in the left lower lobe. A diagnosis of adenocarcinoma like bronchioloalveolar carcinoma (BAC) was made for the lesion in the right upper lobe by means of a transbronchial biopsy. However, the specimen from the left lower lobe showed nonspecific changes, and bronchiolitis obliterans organizing pneumonia (BOOP) or a metastatic lesion from the BAC was suspected. We prescribed steroids for the short term. Since the lesion in the left lobe disappeared, we performed a right upper lobectomy. In cases of primary lung carcinoma, BOOP in the opposite lung is rare and it was difficult to decide on the operability. We concluded that steroid therapy is useful for distinguishing between inflammatory and tumorous lesions in cases in which a detailed examination is difficult. PMID- 12642918 TI - [Mycoplasma pneumoniae pneumonia with a tumorous shadow on chest radiography--a case report]. AB - A 16-year-old woman was admitted with a 1-week history of increasing cough and fever. Chest radiography and computed tomography revealed a tumorous shadow in the right upper lung field. Serologic testing provided evidence of a Mycoplasma pneumoniae infection. The clinical findings did not improve after 3 days of minocycline treatment, or after another 3 days in which levofloxacin was administered in combination with minocycline. The patient gradually improved after treatment with erythromycin replaced the minocycline treatment. This was a rare case of Mycoplasma pneumoniae pneumonia with a tumorous shadow in the chest radiograph. PMID- 12642917 TI - [A case of diffuse malignant mesothelioma of the pleura with bilateral chylothorax]. AB - We describe a case of bilateral chylothorax with malignant pleural mesothelioma. A 41-year-old woman was admitted to our hospital because of dyspnea. She had no history of exposure to asbestos. A chest radiograph and chest computed tomogram (CT) on admission revealed massive bilateral pleural effusion and a large tumor with pleural thickening in the left lateral and anterior parts of the pleura and mediastinum. Biochemical tests of pleural fluid revealed chyle. Two years before, she had been diagnosed through histological and histochemical examinations as having diffuse malignant pleural mesothelioma of the epithelial type. Chest-tube drainage was performed, and pleurodesis was induced by the intrathoracic injection of OK-432 at 10 KE per dose. The chylothorax disappeared after pleurodesis. To date, reports of malignant mesothelioma with nontraumatic chylothorax have been rare. PMID- 12642919 TI - [A case of pulmonary leiomyosarcoma]. AB - A 73-year-old woman with hemoptysis visited our hospital. Chest radiography showed a massive shadow on the right middle lung field. Bronchofiberscopic examination demonstrated a polypoid tumor obstructing the right middle lobe bronchus. A chest CT scan showed that the tumor was situated in the right middle lobe, obstructing the right pulmonary artery trunk. Sarcoma was diagnosed after a CT-guided biopsy. The tumor grew rapidly, giving rise to brain metastasis, which led to the death of the patient. An autopsy examination confirmed the diagnosis as pulmonary leiomyosarcoma. PMID- 12642920 TI - [Women and diabetes]. AB - The objective of the work was to assess, based on the results of studies conducted abroad and the author's own results from the Prague register of diabetic patients, whether there exist differences in the epidemiology, clinical course of diabetes mellitus (DM) and its complications between female and male diabetics. As far as type 1 DM is concerned there are significant differences in the clinical course of diabetes, i.e. deteriorated compensation of DM in girls and women in conjunction with menstruation and the menopause. As compared with men, young women have a higher mortality on account of diabetic nephropathy, and at any age they have as higher incidence of hypertension. In DM2 they have also a high incidence of hypertension and risk of compensation of DM during menstruation and later during the menopause. A significantly less frequent complication in women is ischaemia of the lower extremities. From the epidemiological aspect a higher prevalence of DM2 was proved in women after the age of 65 years. PMID- 12642921 TI - [Biochemical changes in pregnant diabetics]. AB - In the course of pregnancy increasing insulin resistance develops due to deteriorated phosphorylation of the insulin receptor and its substrate (IRS-1). As a result of relations between secretion and the action of insulin the beta cell reacts by insulin hypersecretion and thus the development of hyperinsulinaemia is influenced. Furthermore pregnancy is characterized by increasing differences between the anabolic and catabolic stage in the intermediary metabolism manifested in particular by a tendency towards ketosis and hypoglycaemia during fasting and also by hypoaminoacidaemia. In addition to these metabolic changes there are also other laboratory deviations, manifested e.g. by hyperfibrinogenaemia and an increased concentration of tPA inhibitor (PAI 1) without detectable endothelial dysfunction. The increasing albuminuria is not a manifestation of developing nephropathy but the sign of functional changes which after delivery disappear very quickly. The mentioned deviations are the consequence of altered hormonal regulations in the course of pregnancy, while after delivery the baseline state present before pregnancy is restored. PMID- 12642922 TI - [Pregnancy and labor in diabetes from the viewpoint of the obstetrician]. AB - Diabetes of all types is a known complication of pregnancy and is a risk for mother and foetus. Despite significantly improved care of pregnancies complicated by diabetes the perinatological results are still somewhat worse than in the healthy population. Therefore in the further improvement of perinatological indicators preconception care participates as well as systematic compensation of insulin dependent diabetes, preventive admission on hospital and correct timing of termination of pregnancy in pregestational as well as in gestational diabetes. All care of these pregnancies is shared by the obstetrician and diabetologist. PMID- 12642923 TI - [Views of a diabetologist on pregnancy in diabetic patients]. AB - In recent years a very marked drop of complications in mothers with type 1 diabetes (DM 1) and type 2 diabetes (DM 2) was achieved. Nevertheless this group of patients is still considered at risk. The most serious possible complications are congenital malformations of the foetus and possible deterioration of long term diabetic complications of the mother. The subsequent text is a brief review of contemporary views on pregnancy of diabetic patients. PMID- 12642924 TI - [Gestational diabetes mellitus]. AB - Any degree of carbohydrate intolerance which develops for the first time during pregnancy is defined as gestational diabetes mellitus (GDM). GDM is often discovered late, in more than half the cases only during complications associated with pregnancy. Early diagnosis of GDM and early correct therapy of GDM leads to reduction of complications associated with pregnancy, delivery and the perinatal period of the infant. As in our country unequivocal rules for detection of gestational diabetes are still lacking. We feel that introduction of correct routine screening will contribute to the earlier diagnosis of GDM in all women and thus also to appropriate therapy of gestational diabetes. PMID- 12642925 TI - [Preconception care of the type I diabetic patient from the viewpoint of the diabetologist]. AB - High standard metabolic compensation of diabetes reduces in a significant way the risk of developmental defects and the perinatal morbidity of children of diabetic mothers and the maternal mortality. A considerable proportion of diabetic women is inadequately informed on the risks of pregnancy. As many as two thirds of pregnancies in diabetic women are unplanned and thus the risk of inadequate compensation of diabetes in early stages of pregnancy increases. Preparation of diabetic women for planned pregnancy is therefore very important. Ensuring long term normal blood sugar levels in type 1 diabetics has many pitfalls and involves in particular optimal insulin treatment and systematic and high standard control of the blood sugar level and other parameters. PMID- 12642927 TI - [Urogenital infections in women with diabetes]. PMID- 12642926 TI - [Gynecologic neoplasms in diabetic patients]. AB - Gynecologic malignancies (including breast cancer) represent a substantial proportion of neoplastic disorders in women. The problems associated with the treatment of gynecologic cancer is not limited to gynecology and oncology, but involve other specialties, including, in the case of breast cancer, surgery and radiology. The incidence of gynecologic cancer increases with age, similarly to other tumors or some internal disorders, including diabetes mellitus. In many patients, especially elderly, a coincidence of cancer and diabetes mellitus is observed. The presence of diabetes mellitus, similarly to that of other comorbid conditions, may have a profound impact on the prognosis and the choice of treatment for the individual patient. Moreover, some studies indicate that diabetes mellitus increases the risk of breast and endometrial carcinoma. As in other areas of medicine, a close collaboration between specialists treating the tumor (surgeon or gynecologist, medical and radiation oncologist) with physicians specialized in treating comorbid conditions (internal medicine, cardiology or diabetes medicine) is inevitable. The current state of the treatment of gynecologic cancer is reviewed, with a special focus on breast cancer. The progress in breast cancer treatment illustrates how the understanding of molecular mechanisms underlying tumor growth and evidence-based medicine can lead to a major improvement of the prognosis of cancer. PMID- 12642928 TI - [Anesthesia in labor and gynecologic procedures in women with diabetes]. PMID- 12642929 TI - [Nutrition in women with diabetes in various stages of life]. AB - The diet of diabetic women should not differ fundamentally from an ordinary healthy diet. The intake of the basic energy providing nutrients and of microelements should be balanced and if possible should be supplied in natural food products. This ensues among others from the fact that the relations between some micronutrients are not yet quite clear. During pregnancy it is however necessary to ensure in diabetic women 400 micrograms folic acid; increased attention must be paid to magnesium intake and during the menopause the vitamin D and calcium intake must be followed up. PMID- 12642930 TI - [Metabolic aspects of the polycystic ovary syndrome]. AB - The author reviews the history of the term polycystic ovaries syndrome. He emphasizes the importance of insulin resistance in this disease. According to more recent criteria for the definition of the syndrome suffices the finding of hyperandrogenism, an irregular cycle (after elimination of other classical causes of this condition) and insulin resistance. The frequency of the disease varies in different populations up to 10%. It is significantly associated in particular with type 2 diabetes and obesity. The molecular biology of the syndrome is obscure. The metabolic syndrome as well as the polycystic ovaries syndrome have partly a genetic pathogenesis as well as an environmentally induced participation caused by stress. The polycystic ovaries syndrome is nowadays unequivocally an atherogenic syndrome and is a unit very close to Reaven's metabolic syndrome X or is part of this syndrome. PMID- 12642931 TI - [Sexual problems in women with diabetes mellitus from the viewpoint of the diabetologist]. AB - Diabetes mellitus, as every chronic disease, leads to the development of a number of complications. The author pays attention to sexual aspects of this disease in women from the aspect of the diabetologist. PMID- 12642933 TI - [Gynecologic complications in obesity]. AB - The author presents a review of gynaecological complications of obesity. He mentions the problem of gestation in obesity, surgical complications and hormonal abnormalities in obese women. A classical complication of obesity is sterility. Obesity is also a risk factor for a number of gynaecological tumours. The author emphasizes the importance of weight reduction in the treatment of sterility and for the prevention of gynaecological tumours. An increase of body weight must be prevented in gynaecological hormonal treatment and contraception. PMID- 12642932 TI - [Urinary incontinence and diabetes mellitus in women]. PMID- 12642935 TI - [Contraception in female diabetic patients]. PMID- 12642934 TI - [Hormone replacement therapy in female diabetic patients]. PMID- 12642936 TI - [Effect of hormonal changes in women on hemostasis]. AB - Oral contraception and hormonal postmenopausal treatment is used worldwide by more than 100 million women. Since 1960 it is known that female sex hormones increase the risk of venous thrombosis, myocardial infarction and acute cerebrovascular attacks. This risk diminishes when small doses of oral contraceptives are administered but it still remains the main cause of thromboembolism in young women. The risk is greatest during the first year of contraceptive use, in women taking desogestrel and gestoden (so-called third generation progesterones) and in women with a prethrombotic predisposition. Hormonal substitution treatment (HRT) increases the risk of venous thrombosis, in particular in the first year of HRT use. In women pregnancy and the puerperium are risks for the development of venous thromboembolism. PMID- 12642937 TI - [Broad-range fungal PCR in diagnosis of complicated infective endocarditis]. AB - Infective endocarditis (IE) frequently causes problems in diagnosis, especially where blood cultures are negative and when IE has fungal etiology. The aim of this study was to evaluate the usefulness of broad-range fungal PCR in diagnosis of fungal super-infection of bacterial IE. Twenty five blood samples were taken for analysis from patients with infective endocarditis. IE was diagnosed according to Duke criteria including positive blood cultures. Suspicion of fungal superinfection was established in 5 patients on serological investigation and confirmed by blood culture in 2 patients. Control group consisted of 15 patients without infection. DNA was isolated using the commercially available S.N.A.P. kit. Amplification products were analyzed by gel electrophoresis. Fungal DNA was found in 2 patients with fungal super-infection of bacterial IE confirmed by culture. In the remaining patients with IE and controls no fungal DNA was found. CONCLUSION: Broad-range fungal PCR is a fast and inexpensive tool for the detection of fungal DNA. The method may be valuable in the diagnosis of fungal super-infection of bacterial IE. PMID- 12642939 TI - [Increased serum levels of vitamin D and calcium in young men after replacement of butter with soft margarine in usual diet]. AB - Osteoporosis features include reduction of bone mass and increased predisposition to fractures. Osteoporosis is considered as one of frequent diseases developing in menopausal women and old men. The values of peak bone mass are the determinant of osteoporosis development. It is estimated that the peak bone mass is reached at the age between 16 and 30 years. (NIH Consensus, 2001). Insufficient intake of calcium and vitamin D in diet, with reduced sun exposure, low physical activity, low body weight are well documented risk factors for this disease in human. For that reason, in osteoporosis prevention it should be tried to ensure adequate supply of calcium in diet from early life, with adequate supply of vitamin D, as factors determining adequate mineralization of osseous tissue. The study was carried out for assessing changes of the levels of calcium and 25-OH cholecalciferol in serum replacing butter in usual diet with margarine enriched with vitamin D. After a period of diet stabilization the study group received first diet with butter, followed by diet with margarine in place of butter. Throughout the whole experiment the diet contained unchanged amounts of dairy products as a source of alimentary calcium. In the experiment 41 young healthy men aged 22.6 +/- 1.2 years participated. Every subject consumed in a period of 4 weeks 30 g of butter daily, and during the following 4 weeks margarine 30 g daily. Substitution of margarine for butter in usual diet raised 25-OH cholecalciferol level in serum by 5.56 ng/ml, that is 32.4% (p < 0.001), with consequent 0.12 mg (5%) rise of serum calcium level (p < 0.05). The intake of calcium with diet from dairy products was unchanged throughout the experiment 576 581 mg daily, that is 72% of the recommended intake. The obtained results suggest that independently of the amount of vitamin D intake with food and calcium with diet substitution of margarine enriched with vitamin D for butter was beneficial raising serum vitamin D level, and at the same time increasing calcium absorption from food. It can be accepted that the obtained results could support the opinion on beneficial effects of the consumption of high-quality margarine in daily diet as prevention of the development not only of atherosclerosis but also osteoporosis. PMID- 12642938 TI - [Comparison of interleukin-6 and C-reactive protein serum concentrations assessment in diagnosis of infective endocarditis]. AB - Serum interleukin-6 (IL-6) level might be used to aid in diagnosis of infective endocarditis (IE), especially when blood cultures are negative. One of typical acute phase proteins is C-reactive protein (CRP), often served as an additional inflammation maker. The aim of the study was to compare serum IL-6 and CRP concentrations assessment in diagnosis and monitoring of IE. The study group consisted of 40 patients with ongoing IE and valvular heart diseases. Two control groups consisted of patients with valvular heart diseases: 15 without infection and another 15 with urinary tract infection. The diagnosis of IE was established according to the Duke University criteria; in 34 patients positive blood cultures were found. Serum IL-6 and CRP were measured on three occasions after diagnosis of IE was established and during antimicrobial treatment (mean period 14 +/- 7 days) by ELISA. Usefulness of both parameters for IE diagnosis were described. Reference values were defined as 0-12.5 pg/ml for IL-6, and 0-10 mg/l for CRP. Mean concentrations of IL-6 and CRP in patients with IE (37 +/- 44.3 pg/ml and 27.1 +/- 23.9 mg/l) were significantly higher than in controls: with urinary tract infection (9.1 +/- 4.42 pg/ml and 6.95 +/- 4.39 mg/l) and without infection (3.95 +/- 1.4 pg/ml and 2.21 +/- 0.96 mg/l). CRP concentration was not significantly correlated with IL-6 (r = 0.51, p = 0.07). The significant tendency of consecutive IL-6 concentrations to decrease (from 37 +/- 44.3 to 8.7 +/- 5.7 pg/ml), without decrease of CRP (from 27.1 +/- 23.9 mg/l to 22 +/- 18.3 mg/l) was found. CONCLUSIONS: 1. Elevated serum IL-6 and CRP levels may suggest ongoing IE. 2. Sensitivity, specificity, positive and negative predictive value are nonsignificantly higher for IL-6 than CRP. 3. Combined assessment of serum IL-6 and CRP concentration has no higher value in diagnosis of IE. 4, IL-6 level decrease is faster than CRP during antimicrobial treatment, and it helps better and faster monitoring of treatment. PMID- 12642940 TI - [Short term stimulation of megakaryopoiesis in cord blood derived hematopoietic stem cells in ex vivo model]. AB - Ex vivo expansion of megakaryocytes may be useful to prevent the long-term severe thrombocytopenia following umbilical cord blood transplantation. This study aimed to establish the optimal conditions of short-term stimulation of megakaryopoiesis. Immunomagnetically isolated CD34 cells of umbilical cord were placed into serum free culture medium supplemented with combinations of growth factors as follow: Tpo (thrombopoietin) and IL-3 (interleukin 3); Tpo, IL-3 and SCF (stem cell factor) or Tpo, II-3, SCF and IL-11 (interleukin 11). The number of colonies of megakaryocytic cells (expressed CD41 antigen) were evaluated in clonal culture in 24-hours intervals up to 5 days. Besides the influence of used culture systems on the other hematopoietic lines was checked on the same time points. It was shown that the best results were obtained using combination of 4 cytokines. At 5-th day of expansion the number of CD41 positive cells increased in this warrant 8-fold. In case of erythroid or granulocyte-macrophage precursors the use of this combination of growth factors resulted in slightly elevated number of them. We can conclude that proposed model of expansion of megakaryopoiesis seems to be very effective and can to be useful for the shortening the post transplant thrombocytopenia period. PMID- 12642941 TI - [Serological tests for detection of fungal infection connected with prolonged antibiotic treatment after valve replacement]. AB - In recent years substantial progress has been seen in the development of approaches and methods for serological diagnosis of the mycosis. The aim of our study was to assess the clinical usefulness of serological tests for fungal infection in pts who underwent prolonged antibiotic therapy after valve replacement. MATERIAL AND METHODS: The group consisted of 27 pts after valve replacement with prolonged antibiotic therapy (6-12 weeks) due to infective endocarditis (22 pts) or sternal infection (5 pts). The samples of blood were taken 10 days after antibiotics were started and next every 10 days. Serological techniques for the diagnosis of Candidiasis consisted of antibody detection to: the cell wall polysaccharide antigen (test A), cytoplasmic protein antigen (test B) and the germ tube antigens (test C). RESULTS: Patients were divided in 3 groups due to the kinetics of the antibody responses to the Candida antigen: 1. pts with normal level of antibody during antibiotic treatment--they didn't required antifungal therapy, 2. pts with increasing level of antibody needed antifungal treatment, 3. pts with increasing kinetics of antibody responses during antibiotic and antifungal treatment needed the change of antifungal therapy, 4. Control group (pts before cardiac surgery without any sign of fungal infection). Antibody to the cytoplasmic antigen was detected infrequently. The presence of antibody to the cytoplasmic antigen was correlated with the patient clinical outcome and prognosis and necessity for change of antifungal therapy. One pt died after valve replacement due to valve dysfunction because of fungal endocarditis. CONCLUSIONS: Serological tests for Candidiasis can be a useful marker of fungal infection, prognosis and treatment in pts required prolonged antibiotic therapy after valve replacement. PMID- 12642942 TI - [The whole blood and plasma viscosity changes in course of acute myocardial infarction]. AB - The whole blood and plasma viscosity changes in course of acute myocardial infarction were examined. The examination were performed at the beginning of acute phase of myocardial infarction (period 1), at second to third day (period 2) and after about 10 days of infarction episode (period 3). 77 patients (mean age 56.8 +/- 9.8 years) suffered from myocardial infarction were examined. The whole blood viscosity at following shear rates [s-1]: 0.116; 1.0; 4.59; 150 and plasma viscosity were performed. Besides the viscometric examinations the total cholesterol, LDL-cholesterol, HDL-cholesterol, triglycerides, glucose and fibrinogen as well as blood morphology and ESR were determined. All rheological measurements were carried out at the temperature of 37 degrees C immediately after blood drawing. The control group consisted of 110 healthy persons (aged 56.6 +/- 10 years). Some persons of control group have got risk factors of atherosclerosis as: obesity, artery hypertension and cigarette smoking. The following additional parameters were investigated: hematocrit, the artery pressure, the body mass index, total cholesterol concentration, serum LDL cholesterol, HDL-cholesterol, fibrinogen and blood morphology. The corrected whole blood viscosity was adjusted to 45% of hematocrit. It was stated that the native whole blood viscosity was disturbed at all periods of disease. The corrected whole-blood viscosity in all periods of acute myocardial infarction comparing with controls increased. The greatest rise of corrected whole blood viscosity was especially observed in second period of acute myocardial infarction. Plasma viscosity in patients with acute myocardial infarction is increased in all periods. The greatest rise of plasma viscosity was in second period of disease. The rheological blood and plasma disturbances were connected with increase of total cholesterol, LDL-cholesterol, triglycerides and fibrinogen. These disturbances of blood and plasma viscosity may play a role in promoting myocardial infarction factors. PMID- 12642943 TI - [Reversible thyrotoxic cardiomyopathy--report of three cases]. AB - Three patients with thyrotoxicosis, atrial tachyarrhythmia and congestive heart failure despite successful treatment of hyperthyroidism revealed atrial fibrillation/flagellation and left ventricular systolic dysfunction. Congestive heart failure resolved and left ventricular systolic function normalized only after successful cardioversion to sinus rhythm. In some patients treated for hyperthyroidism, achievement of euthyroid state is not by itself sufficient to reverse left ventricular failure. Improvement after successful reversion of atrial tachyarrhythmia suggest its essential role in pathogenesis of thyrotoxic cardiomyopathy. PMID- 12642944 TI - [Insulinoma of the pancreas--difficulties with imaging]. AB - We present a case of 29-year old man in whom despite the lack of finding tumor in USG and CT surgical intervention was performed and insulinoma of the pancreas was excised. Insulinoma was diagnosed on the basis of clinical picture (Whipple's triade) that was reassured by positive fast test. The fast was terminated at 11 and half hours because of neuroglycopenic symptoms with serum glucose and insulin levels 2.0 mmol/l (36 mg/dl) and 11.07 microU/ml respectively, which established the diagnosis. Laparotomy was performed, during which the tumor was palpated and seen in ultrasonography in the head of pancreas. The tumor of 12 mm diameter was excised. Histopathological examination revealed benign insulinoma. After surgery the symptoms alleviated completely. The patient with proper glucose levels was discharged in good health. The case supports high usefulness of intraoperative ultrasonography and surgeon's palpation. PMID- 12642945 TI - [Role of TGF-beta in the pathogenesis of atherosclerosis]. PMID- 12642946 TI - [Cytogenetics and molecular genetics of non-Hodgkin's lymphoma]. PMID- 12642947 TI - [Raynaud's disease and Raynaud's phenomenon in connective tissue diseases]. PMID- 12642948 TI - [Antihypertensive treatment in patients with diabetes mellitus]. PMID- 12642949 TI - Auxiliary nurse midwives vocational course--a study. PMID- 12642950 TI - Nurses promote positive mental health. PMID- 12642951 TI - Dental Council of New Zealand decisions on specialist registration issues. PMID- 12642952 TI - Dens evaginatus--how does it present and how should it be managed? AB - Dens evaginatus is a developmental dental anomaly predominantly affecting premolars in people of Mongolian origin. It is likely to become more prevalent in New Zealand due to the increase in immigration of Asians. The formation of dens evaginatus is postulated to be an abnormal evagination of the internal enamel epithelium and dental papilla into the stellate reticulum during morphodifferentiation. An important complication resulting from damage to the evagination is periapical abscess. When considering treatment options, the long term prognosis of the tooth, its root length, the general alignment of the teeth, and overall arch length should be reviewed. Management options include composite resin placed at the base of the tubercle, excision of the tubercle followed by direct or indirect calcium-hydroxide pulp capping and composite-resin restoration, and apexification procedures followed by conventional endodontic therapy. Grinding of the intact tubercles of newly erupted evaginated premolars may contribute to secondary infection of the pulp, and is not recommended. PMID- 12642953 TI - Clinical aspects of resin-bonded bridges. AB - Resin-bonded bridges have emerged as an attractive, minimally invasive prosthodontic option for the restoration of edentulous spaces. Although success rates, determined by retrospective studies, vary greatly, resin-bonded bridges now have success rates approaching those of their cemented counterparts; success is dependent, however, upon appropriate design and careful technique. The metal framework must be rigid and of sufficient surface area to afford secure adhesion to the underlying teeth. Tooth preparation should be restricted to enhancing the bonding area, and it is important that prostheses are designed to maintain axial loading, and guidance upon metal. PMID- 12642954 TI - AIDS in New Zealand. PMID- 12642955 TI - General Shrapnel and the "unfortunate" Dr. H. J. Shrapnell. AB - Several aspects of Shrapnell's career are still little known or erroneously typed. 1. Some historians of Medicine profess that Dr. Shrapnell was the inventor of a shell which bears his name. 2. Others neglect the negative points of Shrapnell's paper published in 1832 on the pars flaccida. To be able to comment on these different aspects of Shrapnell's life and work we compared the biographies of the anatomist and of the artillery officer. We noticed the positive and also the negative points of his paper. We conclude that: 1. Dr. Shrapnell was not an artillery officer, but the two men were present in the British Army; their surnames, their names and the period when they lived were about the same. 2. Errors in Shrapnell's paper were due to the philosophy and lack of anatomical and pathological knowledge of the time. 3. Otologists' reception was better than that of the Anatomists because for the Otologists, Shrapnell's membrane was recognized as an important original area of cholesteatoma, whereas for the Anatomists it was just a detail. PMID- 12642956 TI - The influence of oral implant-supported prostheses on articulation and myofunction. AB - The aim of the present research was to assess articulation and myofunction in patients wearing fixed or removable prostheses supported by oral implants. 164 edentulous patients with implant supported prostheses were divided in four subgroups, dependent on their dental status, and compared to control groups of forty five subjects having a natural dentition. More than fifteen articulatory and myofunctional parameters were evaluated. The results showed that subjects with prostheses on implants tend to have more articulation problems than controls. Especially patients with a complete fixed prosthesis on implants in the upper jaw seemed to experience problems pronouncing /s/ and /z/. There also seemed to be influences of age and hearing factors. PMID- 12642957 TI - Malignant fibrous histiocytoma of the pharynx. AB - Malignant fibrous histiocytoma (MFH) is the most common soft-tissue sarcoma of late adult life, but is relatively uncommon in the head and neck region. That region has been reported to be the origin of malignant fibrous histiocytoma in 3 10% of cases. Only one case of the tumor occurring in the pharynx has been reported. Histologically it is sometimes hard to distinguish this tumor from some sarcomas and pleomorphic carcinomas. The treatment of choice is a large surgical resection, while radiotherapy and chemotherapy are reserved for recurrences. The authors present a case of oropharyngeal malignant fibrous histiocytoma. The patient complained dysphagia and dyslalia progressively worsening in six months. Pharyngo-laryngoscopy revealed a mass of the left lateral wall of oro and hypopharynx. CT scan examination showed a capsuled mass which displaced but not involved the neck neurovascular structures; there was no evidence of linphonodal involvement. Transoral surgical excision of the mass was performed with the preservation of speech and swallowing. For more than 1 year postoperatively, there has been no evidence of the disease or metastasis. PMID- 12642958 TI - Systemic leishmaniasis involving the nose. AB - A case of a patient, presenting with a granulomatous lesion of the anterior nasal septum mucosa spreading to the columella and the nasal floor, whereby leishmaniasis was diagnosed, is presented. The clinical and pathological aspects of this pathology, its diagnosis and treatment are reviewed. PMID- 12642960 TI - Healing of maxillary sinusitis of odontogenic origin following conservative endodontic retreatment: case reports. AB - Because of the anatomical proximity, infection of posterior upper teeth can spread into the maxillary sinus. When confronted with a large persistent periapical lesion on a posterior maxillary root-treated tooth, the practitioner should question the quality of the previous endodontic treatment, evaluate the impact of the potential causes of failure and consider, in the majority of cases, the conservative non-surgical retreatment instead of extraction or periapical surgery of the dental element. This paper reports two cases of healing of extensive periapical bone destruction and of the co-existing sinus pathology of odontogenic origin after non-surgical endodontic retreatment of previously root treated upper molars. Misconceptions concerning the role of epithelium in the periapical lesion are discussed. Also, emphasis is put on the need of precise radiological diagnosis, pre-operatively as well as post-operatively to ascertain healing and to avoid unnecessary delay in the appropriate management of these patients. PMID- 12642959 TI - Aggressive psammomatoid ossifying fibroma of the inferior turbinate and lateral nasal wall. AB - OBJECTIVE: Fibroosseous lesions are rare entities of the nose and paranasal sinuses, the terminology and classification of which are still confusing. Psammomatoid or aggressive (juvenile) ossifying fibroma is a benign messenchymal tumor usually met in the young age (5-15 years), in the sinonasal tract, bearing distinctive histomorphologic features and a tendency towards locally aggressive behaviour. METHODOLOGY: We report here a rare case of an aggressive psammomatoid ossifying fibroma of the inferior turbinate and the lateral nasal wall, with obstruction of the nasolacrimal duct, in a 68-year-old woman. RESULTS: Diagnosis was based on physical examination, CT scan imaging and histopathological examination. Treatment consisted of endoscopic intranasal resection of the tumor accompanied by removal of the lateral nasal wall. CONCLUSIONS: Fibro-osseous tumors of the nose and paranasal sinuses require aggressive surgical approach in order to avoid recurrence. Complete surgical excision may not always be possible mainly due to the tumor's extent and location. PMID- 12642961 TI - Synthesis and biological evaluation of 16 beta-pyrrolidinosteroidal derivatives. AB - The synthesis of some steroidal bisquaternary ammonium substances (compounds 10 and 11), and their in vitro and in vivo neuromuscular blocking action are described in this report. The pyrrolidino functionality was incorporated at both the 3-beta and 16-beta positions of the steroid nucleus to study the importance of the interonium distance between the two quaternary ammonium heads. The 16-beta pyrrolidino monoquaternary derivatives (compounds 14, 15, 16) were also prepared. The 17-beta-acetoxy bisquaternary derivative compound 11 was found to be more potent than d-tubocurarine (CAS 57-94-3) in in vivo studies. The 17-beta-hydroxy bisquaternary derivative, compound 10, and its 16-beta-pyrrolidino monoquaternary partner, compound 15, were found to be less active as compared to d-tubocurarine in in vitro studies. The monoquaternary compounds 14 and 16 were not tested due to their solubility problems. The intermediate substance, compound 12 was selected by the National Cancer Institute (NCI), Bethesda (USA) for investigation for antineoplastic activity but was found to be inactive. PMID- 12642962 TI - Pharmacokinetics and metabolism of radiolabelled SNI-2011, a novel muscarinic receptor agonist, in healthy volunteers. Comprehensive understanding of absorption, metabolism and excretion using radiolabelled SNI-2011. AB - The pharmacokinetics and metabolism of SNI-2011 ((+/-)-cis-2-methylspiro[1,3 oxathiolane-5,3'-quinuclidine]monohydrochloride hemihydrate, cevimeline, CAS 153504-70-2), a novel muscarinic acetylcholine receptor agonist developed for the treatment of Sjogen's syndrome, were investigated in six healthy volunteers after a single oral administration of 14C-SNI-2011. After administration, plasma concentrations of the radioactivity and SNI-2011 reached to Cmax at approximately 2 h, and then decreased with t 1/2 of 9 and 4 h, respectively. Cmax and AUC0 infinity of the radioactivity in plasma were 2.2 and 5.0 times higher than those of SNI-2011, respectively. The main excretion route of the radioactivity was urine, and 97.3% of the dose excreted in urine within 168 h, indicating that 14C SNI-2011 was completely absorbed. The mean recoveries of the metabolites in urine at 24 h after administration were 16.0% for SNI-2011, 35.8% for SNI-2011 trans sulfoxide (SNI-t-SO), 8.7% for SNI-2011 cis-sulfoxide, 4.1% for SNI-2011 N-oxide, furthermore, two unknown metabolites, UK-1 and UK-2, were detected 14.6% and 7.7%, respectively. LC/MS analysis and hydrolysis studies revealed that UK-1 and UK-2 were glucuronic acid conjugates of SNI-2011 and SNI-t-SO, respectively. PMID- 12642963 TI - Bioavailability of opipramol from a film-coated tablet, a sugar-coated tablet and an aqueous solution in healthy volunteers. AB - Opipramol (4-[3-(5H-dibenz[b,f]-azepine-5-yl)-propyl]-1-piperazine-ethanol dihydrochloride, CAS 315-72-0) is regarded as an anxiolytic compound with antidepressant properties, and it is one of the most frequently prescribed psychotropic drugs in Germany. In two open, randomized cross-over studies in 20 (study 1) and 18 (study II) healthy volunteers, the relative bioavailability of 50 mg opipramol-2HCl from a sugar-coated tablet was compared with an aqueous solution, and of 100 mg opipramol-2HCl from a newly developed film-coated tablet was compared with the sugar-coated tablet. The concentrations of opipramol were determined in plasma by high-performance liquid chromatography (HPLC) with photometric detection. The mean dose corrected kinetic parameters of opipramol were similar after administration of all formulations. The peak concentrations of opipramol were 13-15 ng ml-1 (study I) and 28 ng ml-1 (study II). They were achieved after 3 h. The area under the plasma concentration-time curve was about 170 ng ml-1 h (study I) and about 320 ng ml-1 h (study II). The terminal plasma half-life was 11 h. Bioequivalence was proven between sugar-coated tablet and aqueous solution, and between film-coated tablet and sugar-coated tablet, respectively. In addition, in study II the plasma concentrations and pharmacokinetic parameters of the metabolites opipramol N-oxide and deshydroxyethyl opipramol were determined. PMID- 12642964 TI - Comparative antiallergic effects of second-generation H1-antihistamines ebastine, cetirizine and loratadine in preclinical models. AB - Ebastine (CAS 90729-43-4), cetirizine (CAS 83881-51-0) and loratadine (CAS 79794 75-5) are second generation H1-antihistamines of proven efficacy for treating allergy. Recent clinical studies have found ebastine to be more effective than cetirizine or loratadine in alleviating the symptoms of seasonal allergic rhinitis. The objective of this study was to compare the efficacy of these compounds in three guinea-pig modeles of bronchoconstriction, elicited either by histamine, allergen or leukotriene C4 in order to shed light onto the mechanisms that might explain differences found in clinical studies. In the present experiments, ebastine and cetirizine were equipotent against aerosol histamine induced bronchospasm in guinea pigs (ED50 115 and 100 micrograms/kg p.o., respectively), while loratadine was three-fold less potent. In the same model the effects of ebastine, loratadine and cetirizine lasted 21, 19 and 15 h, respectively. Ebastine (ED50 334 micrograms/kg p.o.) was the most potent compound in inhibiting allergen-induced bronchospasm in conscious guinea pigs. In vitro studies in tracheally perfused guinea pig lungs demonstrated that ebastine and loratadine inhibited with equal potency the bronchoconstriction induced by leukotriene C4 whilst cetirizine was significantly less potent. Finally, in another in vivo study, ebastine reverted the changes in pulmonary resistance induced by leukotriene C4 in anaesthetised guinea pigs, whereas cetirizine and loratadine were devoid of activity in this model. In accordance with the clinical data, ebastine proved to be the substance with the widest range of application in animal experiments, too. PMID- 12642965 TI - Pharmacological profile of AS-9705, a novel benzotriazolecarboxamide derivative, as a gastroprokinetic agent with potent anti-emetic activity. AB - The pharmacological profile of AS-9705 ((R)-N-(1-ethyl-1H-hexahydroazepin-3- yl) 6- methoxy-1H-benzotriazole-5-carboxamide fumarate monohydrate, CAS 219622-61-4), a novel gastroprokinetic agent with potent anti-emetic activity, was investigated in the present study. AS-9705 inhibited [3H]spiperone binding to human dopamine D2.long receptors, and [3H]R(+)-7-OH-DPAT binding to human dopamine D3 receptors (IC50 values of 58.5 +/- 14.0 and 60.8 +/- 7.8 (nmol/l), respectively) and had negligible affinity (IC50 > 10 mumol/l) for other neurotransmitter recognition sites examined. Moreover, in ferrets or dogs, AS-9705 dose-dependently inhibited emesis induced by R(+)-7-OH-DPAT and apomorphine with ID50 values of 0.05 mg/kg p.o. and 0.04 mg/kg p.o., respectively. AS-9705 dose-dependently enhanced normal gastric emptying and potently inhibited the delay in gastric empting induced by apomorphine, morphine, cisplatin, clonidine and cholecystokinin in rats. Furthermore, in conscious fasting dogs, AS-9705 dose-dependently stimulated gastric motility. In conclusion, AS-9705 is a novel gastroprokinetic agent with potent antiemetic activity and minimal CNS adverse effects and is, therefore, worthy of clinical investigation. PMID- 12642966 TI - Design, synthesis and in vitro cytotoxicity of a cis-dichloroplatinum (II) complex linked to the minor groove binder stallimycin. AB - Two potentially hydrophilic platinum (II) complexes 10 and 11 bound to the minor groove binder stallimycin (distamycin A, CAS 636-47-5) by L-cysteine and D,L-2,3 diaminopropionic acid have been synthesized. The in vitro cytotoxicity of both these complexes was evaluated against several cell lines. None of the synthesized platinum complexes showed greater activity than that of cisplatin (cis-DDP, 1) (CAS 15663-27-1). Interestingly, the free ligands 6 and 9 were more active than the related platinum complexes 10 and 11, respectively, with respect to RAJI, CCRF-CEM and MOLT-4 human leukaemia cell lines. PMID- 12642968 TI - Developmental toxicity studies of the quinolone antibacterial agent irloxacin in rats and rabbits. AB - Embryotoxicity studies on irloxacin (6-fluorine-7-(pyrrol-1-yl)-1-ethyl-1,4 dihydro-4-oxo-quinolone-3-carboxylic acid, CAS-91524-15-1), a new fluoroquinolone antibacterial agent, were performed in rats and rabbits. Oral administration of irloxacin during the fetal period of organogenesis to pregnant rats and rabbits at dose levels of up to 1000 and 350 mg/kg/d, respectively, elicited no evidence of teratogenicity. During the first days of treatment, transient stasis in body weight increase was observed in rat dams receiving doses of 350 or 1000 mg/kg/d, and reduced food consumption was observed in those receiving 1000 mg/kg/d. Necropsy on day 20 of gestation showed dosage related increase in liver and kidney weights in all rat treated groups. Rabbit dams receiving 350 mg/kg/d showed during the first days of treatment decrease in body weight, and decreased food consumption and faecal output. Also, four females receiving 350 mg/kg/d aborted between days 18 and 20 of gestation. Rat fetuses in the 350 and 1000 mg/kg/d showed decreased body weight, and a decrease in placental weights was observed in the 1000 mg/kg/d group. No retardations or malformations were observed in rat or rabbit fetuses at any tested dose level. For maternal and embryo-fetal effects 100 and 150 mg/kg/d can be considered as the no-effect-level (NOEL) for rats and rabbits, respectively. PMID- 12642967 TI - Synthesis and evaluation of analgesic, anti-inflammatory and antimicrobial activities of 6-acyl-3-piperazinomethyl-2-benzoxazolinones. AB - In this study, 16 new 6-difluorobenzoyl-3-piperazinomethyl-2-benzoxazolinones were synthesized by Mannich reaction. Their chemical structures were proven by IR, 1H-NMR and elemental analysis. The compounds were screened for their analgesic and anti-inflammatory activities. A modified Koster test, using acetylsalicylic acid (ASA, CAS 50-78-2) as the reference drug, was used to assess analgesic activity. The anti-inflammatory activity was evaluated by the carrageenan induced hind-paw oedema test. The analgesic activities of all compounds were higher than their anti-inflammatory activities and therefore the prominent analgesic actions of the compounds are thought to be due to a central effect. The microbiological effects of the compounds were evaluated in vitro against various pathogenic fungi and bacteria using the microdilution method. Most of the compounds were found to be inactive against bacteria and fungi. One of the compounds (31) possessed considerable analgesic activity as well as moderate antibacterial activity against S. aureus. Another compound (3m) showed analgesic and antifungal activities comparable to those of ASA and fluconazole (CAS 86386-73-4), respectively. PMID- 12642969 TI - Suitability of cefotiam and cefuroxime axetil for the perioperative short-term prophylaxis in tonsillectomy patients. AB - The efficacy of the perioperative short-term prophylaxis with cefotiam (CAS 66309 69-1) and cefuroxime axetil (CAS 64544-07-6) was analysed by the assessment of the pharmacological kinetics in the serum and the tonsil tissue in 50 patients with recurrent tonsillitis. Twenty-four patients received 1 g cefotiam by the intravenous route 30 min to 4 h before the tonsillectomy, and 26 patients received 250 mg cefuroxime axetil orally 1 to 6 h before the tonsillectomy. Bactericidal serum levels were reached for cefotiam up to 4 h after intravenous application and for cefuroxime axetil up to 3 h after oral application. In the tissue of the tonsil there were proved levels which were definitely above the MIC 90 (MIC = minimum inhibitory concentration) known for the clinically relevant germs for cefotiam after 30 min up to 2 h, for cefuroxime axetil after only 2 h. Considering the distribution areas, the capacity of the protein binding and the microbiological measuring methods, one can expect an efficient antibiotic coverage after an intravenous one-shot bolus injection of 1 g cefotiam from 30 min to 4 h and after oral application of 250 mg cefuroxime axetil on an empty stomach from 1 to 6 h. Because of the short duration of a tonsillectomy and the serum and tonsil tissue kinetics cefotiam and cefuroxime axetil are suitable for the perioperative antibiotic prophylaxis of high-risk patients. PMID- 12642971 TI - Toward improved adverse event/suspected adverse drug reaction reporting. PMID- 12642970 TI - Electronic-topological study of the structure-activity relationships in a series of steroids with mineralocorticoid binding affinity. AB - Conformational analysis and quantum chemical calculations were carried out using molecular mechanics (MMP2) and semi-empirical quantum chemistry (CNDO/2) methods for 51 steroid homologues belonging to a series of 17-spirolactones. Matrices called Electronic-Topological Matrices of Conjunction (ETMCs) were formed using data obtained from quantum chemical calculations. A structural fragment of activity was identified in the series of steroids. As seen from the fragment's properties, active compounds are characterized by the presence of two atoms of oxygen, O1 and O3, which are situated at a distance of 13.5 A and possess high negative charges (-0.29 to -0.31 e). PMID- 12642972 TI - Adverse drug reactions, MedWatch reporting and medical student education. PMID- 12642973 TI - Improving the quality of adverse drug reaction reporting by 4th-year medical students. AB - PURPOSE: Evaluate whether a 15-minute lecture intervention will improve adverse drug reaction reporting quality on standard MedWatch forms. METHODS: Seventy eight 4th-year medical students were randomized to intervention 'Group-A' or non intervention 'Group-B' on the first day of a required five-day clinical pharmacology rotation. Group-A participants attended a 15-minute lecture on completing a MedWatch form with quality information considered by the Food and Drug Administration as critical to adequate adverse drug reaction reporting. Group-B participants did not attend this lecture. Both groups then watched a standardized patient interview of a recognizable adverse drug reaction and completed MedWatch forms. Four Safety Evaluators from the Food and Drug Administration (FDA) rated student responses in a blinded fashion for the primary efficacy variable of Overall Impression and six informational domins using a standardized data quality analysis form that was developed within the Office of Postmarketing Drug Risk Assessment of the FDA. RESULTS: Seventy-eight MedWatch forms were evaluated (Group-A = 40, Group B = 38). Overall MedWatch information quality scores for the intervention group were significantly higher than the non intervention group (p < 0.004). CONCLUSIONS: As little as a 15-minute intervention can significantly improve the quality of adverse drug reaction reporting by 4th-year medical students. Academic medical centers should consider incorporating adverse drug reaction reporting curriculum into the clinical training of medical students. PMID- 12642974 TI - Prescriptions of systemic antibiotics for children in Germany aged between 0 and 6 years. AB - Limited information is available about systemic antibiotic use among children in Germany. We therefore assessed prescription patterns by office-based physicians to analyse antibiotic consumption in early childhood. A total of 331 children < 6 years were eligible for inclusion. The number of antibiotic prescriptions, consumed daily doses, number of treatment courses, types of antibiotics and diagnoses for prescribing were determined. The prevalence of systemic antibiotic treatment was 42.9%. Antibiotic consumption was highest between 2 and 3 years of age (55.8%). The percentage of children receiving one, two or three courses of antibiotic treatment was 49.3, 28.2 and 16.2%, respectively. Acute otitis media (32.2%), upper respiratory tract infections (18.9%), tonsillitis (15.9%) and acute bronchitis (15.4%) were principal indications for treatment. Macrolides were most frequently prescribed (48.1%), followed by penicillin V (21.3%), broad spectrum penicillins (14.3%), sulfonamides (10.5%) and cephalosporins (5.8%). Antibiotics not recommended for particular indication were selected in 5-43% of cases. The considerable prescription of systemic antibiotics to children in many European countries is also the case in Germany. A noteworthy trend emerged for suboptimal prescribing with second-line antibiotics. As such treatment may be associated with the development of bacterial resistance, improved guidelines for antibiotic treatment should be drawn up and enforced. PMID- 12642975 TI - Refill compliance in type 2 diabetes mellitus: a predictor of switching to insulin therapy? AB - OBJECTIVE: To assess whether switching to insulin therapy in patients with type 2 diabetes mellitus is associated with medication refill compliance of oral hypoglycemic agents. RESEARCH DESIGN AND METHODS: The PHARMO Record Linkage System was used as data source for this study. Patients with newly treated type 2 diabetes mellitus were defined as subjects in whom oral hypoglycemic therapy was initiated between 1991 and 1998. We performed a matched case-control study in this cohort. Cases were patients who switched to insulin therapy. Date of switching in the case was defined as the index date. Controls were subjects still on oral therapy on the index date, matched on duration of diabetes and calendar time. We measured the medication refill compliance in the year starting 18 months before the index date and calculated various compliance indices. RESULTS: In total, 411 cases and 411 matched controls were identified. Cases suffered more often from more severe comorbidity and used a higher number of oral hypoglycemic agents and concomitant non-diabetic drugs. The overall compliance rate did not differ significantly between cases and controls, the adjusted odds ratio (OR) was 1.3 (CI 95% 0.6-2.8). After performing multivariate logistic regression modeling, age at onset of diabetes, gender, comedication, combination therapy, and daily dosage frequency, were independently related to switching. CONCLUSIONS: We were unable to confirm the hypothesis that noncompliance with treatment is more prevalent in patients with secondary failure. Other variables, like comorbidity and disease-related factors seem to play a more important role in switching to insulin therapy. PMID- 12642977 TI - Drug-induced renal function impairment: a population-based survey. AB - PURPOSE: The knowledge that drugs can affect renal function is mainly based on experimental studies or case reports. Thus, it has only been investigated in selected populations. Here we describe drug groups associated with altered renal function in the general population. METHODS: To study this, we used baseline data of 8592 subjects of a population-based cohort. Hyper- and hypofiltration were defined as a filtration above or below the 90% confidence interval of age-and-sex corrected creatinine clearance. Drug use was measured in the year preceding the kidney function measurement. RESULTS: The prevalence of hyperfiltration (4.6% in the general population) was higher among subjects using anti-diabetics (11.7%), dermatological corticosteroids (5.9%) and sex hormones (5.8%), but lower in subjects using anti-thrombotics (2.4%) and diuretics (2.1%). Hypofiltration (4.3% in the general population) was seen more often in users of beta blockers (6.2%), ACE inhibitors (7.1%), statins (7.2%), anti-thrombitics (6.8%), trimethoprim (7.9%), vaccines (9.4%), NSAIDs (5.2%), anti-ulcer agents (6.3%), laxatives (7.7%) and eyedrops (6.7%). CONCLUSIONS: Several drug groups found in this overview were to be expected, since patients with kidney disease often use them (e.g. cardiovascular drugs). Several other drug groups were somewhat unexpected and deserve further attention. PMID- 12642976 TI - The use of anti-asthmatic medications among pediatric patients in Taiwan. AB - PURPOSE: To describe the use of anti-asthma drugs in Taiwanese pediatric patients. METHODS: Using the data from the database of Bureau of National Health Insurance, we analyze the patterns and extent of medication use among asthma patients less than 18 years old. Drugs were classified according to the Anatomical Therapeutic Chemical (ATC) Classification system. RESULTS: Data on a total of 2,90,467 children were included in the study. Overall, patients were treated more frequently with oral medications. 69.43 and 40.24% of patients were using oral beta-agonists (OBAs) and methylxanthines respectively. Monotherapy with OBAs was the most popular regimen, accounting for 13.70% of the total prescriptions studied. Only 6.70% of patients were taking inhaled corticosteroids (ICSs). During the studied period, users of ICSs were using an average of 1.65 canisters. CONCLUSIONS: The current practices of asthma management in Taiwan fall short of the goals stated in international guidelines. Anti-inflammatory agents were being underused in Taiwanese pediatric patients. PMID- 12642978 TI - For the good of the patient: risks and benefits of medicines. AB - The basis of regulation of medicines is an assessment of risk and benefit. As the process of drug development has evolved with the advances in scientific knowledge, new and more precise methods of measuring drug efficacy have been created. Pharmacovigilance must also change with as much emphasis being placed on demonstrating the safety of a medicine as on preventing harm. Communication of these issues to interested parties remains problematical with room for considerable improvement. PMID- 12642979 TI - Defining dose decrease. PMID- 12642980 TI - Bringing epidemiology into drug utilisation research. PMID- 12642981 TI - Pharmacoepidemiology and drug safety. PMID- 12642982 TI - Changes in dietary patterns and certain nutrition-related diseases in urban and rural residents of Jiangsu Province, China, during the 1990s. AB - OBJECTIVE: To investigate into the changes of dietary patterns and their impacts on health of urban and rural residents in Jiangsu Province, China during the 1990s. METHODS: On the basis of the results of food consumption survey, the calorie intake was calculated according to the balanced diet method and the quality of diet was assessed by scores of desirable dietary pattern (DDP). RESULTS: It was found that food consumption and dietary patterns changed remarkably during the 1990s. Grain consumption was decreasing year by year, but the consumption of animal food was markedly increasing. Although the score of desirable dietary pattern (DDP) in urban residents was more than 90, the deducted score due to over-consumption of animal food increased. The mortality from infectious diseases evidently decreased in the whole province, while the death rates of some chronic diseases, such as diabetes, hypertension, colorectal cancer and breast cancer, were increasing rapidly in urban areas. CONCLUSIONS: Sufficient attention should be paid to the negative effects of change in dietary patterns on people's health, especially in the urban residents. PMID- 12642983 TI - Intake of volatile N-nitrosamines and their ability to exogenously synthesize in the diet of inhabitants from high-risk area of esophageal cancer in southern China. AB - OBJECTIVE: Nan'ao County in Guandong Province is a high-risk area of esophageal cancer in Southern China. Of the suspected etiological factors in the environment, N-nitrosamines and their precursors have received the greatest attention. METHODS: Sixty samples of the diet ingested by the inhabitants were collected and detected for volatile N-nitrosamines and their precursors. Five N nitrosamines detected by Gas Chromatography-Thermal Energy Analyzer were N nitrosodimethylamine, N-nitrosodiethylamine, N-nitrosopyrrolidine, N nitrosopiperidine and N-nitrosomethyl-benzylamine. RESULTS: The average content of 5 volatile N-nitrosamines in the diet was 312.0 micrograms/kg (median). The daily intake of the nitrosamines was 286.5 micrograms/head/day. Only the ability to exogenously synthesize N-nitrosopiperidine was powerful among 5 volatile N nitrosamines. By a computerized stepwise regression analysis and curve fitting, we studied the correlation among the nitrosamines, the precursors and the major food items in the samples. CONCLUSION: It demonstrated that a relatively high content of volatile N-nitrosamines was present in the diet collected in the area. PMID- 12642984 TI - Study of low-intensity 2450-MHz microwave exposure enhancing the genotoxic effects of mitomycin C using micronucleus test and comet assay in vitro. AB - OBJECTIVE: To determine the interaction between 2450-MHz microwaves (MW) radiation and mitomycin C (MMC). METHODS: The synergistic genotoxic effects of low-intensity 2450-MHz microwave and MMC on human lymphocytes were studied using single cell gel electrophoresis (SCGE) assay (comet assay) and cytokinesis blocked micronucleus (CBMN) test in vitro. The whole blood cells from a male donor and a female donor were either only exposed to 2450-MHz microwaves (5.0 mW/cm2) for 2 h or only exposed to MMC (0.0125 microgram/mL, 0.025 microgram/mL and 0.1 microgram/mL) for 24 h; and the samples were exposed to MMC for 24 h after exposure to MW for 2 h. RESULTS: In the comet assay, the comet lengths (29.1 microns and 25.9 microns) of MW were not significantly longer than those (26.3 microns and 24.1 microns) of controls (P > 0.05). The comet lengths (57.4 microns, 68.9 microns, 91.4 microns, 150.6 microns, 71.7 microns, 100.1 microns, 145.1 microns) of 4 MMC groups were significantly longer than those of controls (P < 0.01). The comet lengths (59.1 microns, 92.3 microns, 124.5 microns, 182.7 microns and 57.4 microns, 85.5 microns, 137.5 microns, 178.3 microns) of 4 MW plus MMC groups were significantly longer than those of controls too (P < 0.01). The comet lengths of MW plus MMC groups were significantly longer than those of the corresponding MMC doses (P < 0.05 or P < 0.01) when the doses of MMC were > or = 0.025 microgram/mL. In the CBMN, the micronucleated cell (MNC) rates of MW were 5@1000 and 6@1000, which showed no difference compared with those (4@1000 and 4@1000) of controls (P > 0.05). The MNC rates of 4 MMC groups were 8@1000, 9@1000, 14@1000, 23@1000 and 8@1000, 8@1000, 16@1000, 30@1000 respectively. When the doses of MMC were > or = 0.05 microgram/mL, MNC rates of MMC were higher than those of controls (P < 0.05). MNC rates of 4 MW plus MMC groups were 12@1000, 13@1000, 20@1000, 32@1000 and 8@1000, 9@1000, 23@1000, 40@1000. When the doses of MMC were > or = 0.05 microgram/mL, MNC rates of MW plus MMC groups were much higher than those of controls (P < 0.01). MNC rates of 4 MW plus MMC groups were not significantly higher than those of the corresponding MMC doses. CONCLUSION: The low-intensity 2450-MHz microwave radiation can not induce DNA and chromosome damage, but can increase DNA damage effect induced by MMC in comet assay. PMID- 12642985 TI - Effect of television viewing on pediatric obesity. AB - OBJECTIVE: To test the effect of television viewing on pediatric obesity in urban China. METHODS: Stratified multistage cluster random sampling method was used for subjects selection. Nine thousand three hundred and fifty-six children, as well as their parents, were investigated. Questionnaire survey was used for data collection. Children's weights and heights were measured in the clinic of the investigated kindergartens or schools by trained investigators following the standardized procedure. RESULTS: The percentages of children and adolescents who watched television less than 1 h, 1-2 h, 2-3 h and more than 3 h daily were 32.5%, 46.0%, 15.4% and 6.1%, respectively, while the prevalence of obesity was 10.9%, 11.8%, 13.2% and 15.1%, respectively. Each hourly increment of television viewing was associated with 1%-2% increase in the prevalence of obesity. CONCLUSIONS: Time spent watching television is directly related to an increase risk of obesity, television viewing time is an independent factor for pediatric obesity. PMID- 12642986 TI - Environmental exposure to lead as a risk for prostate cancer. AB - OBJECTIVE: To evaluate the possible role of environmental exposure to lead as a risk factor for prostate pathology in patients suffering from prostate cancer (PCA) and benign prostate hyperplasia (BPH). METHODS: Blood lead (BPb) level was determined in PCA and BPH cases using a graphite furnace Atomic Absorption Spectrophotometer and compared with those in a control group living in the similar socioeconomic environment. RESULTS: BPb was significantly higher in PCA and BPH cases than in normals (P < 0.05). Blood levels of zinc and copper were significantly lower in PCA and BPH cases when compared with controls (P < 0.05). In all the three groups, a statistically significant positive correlation between lead and thiobarbituric acid reactive substances (TBARS) measured as malondialdehyde, and negative correlation between blood lead and antioxidant GSH level, indicative of possible generation of reactive oxygen species, were also observed after adjusting for age as a possible confounders. However, positive association between blood lead and TBARS was relatively higher in PCA patients (r = 0.77, P < 0.05) than in BPH (r = 0.32, P < 0.05) and normal (r = 0.30, P < 0.05). CONCLUSION: These results with limited power seem to suggest for the first time that environmental exposure of aging males to lead may be a risk factor for prostate cancer and/or benign prostate hyperplasia possibly through generation of reactive oxygen species and/or reducing the level of zinc which acts as a cellular growth protector. PMID- 12642987 TI - Mineralization of 14C-ring labelled 2,4-D in Egyptian soils under aerobic and anaerobic conditions. AB - OBJECTIVES: To study the mineralization of 2,4-D in clay and clay loam Egyptian soils under subtropical conditions over a period of 90 d. METHODS: Using 14C-ring labelled pesticide, laboratory studies under aerobic and anaerobic conditions were conducted. 14C-activity in solutions was directly determined by liquid scintillation counting. Unextractable soil residues were determined by combustion. The nature of methanolic 14C-residues was determined by thin layer and high performance liquid chromatographic analysis. RESULTS: Under aerobic conditions 10%-14% of applied dose was mineralized during 90 d irrespective of soil type. The soil extractable pesticide residues decreased with time and the bound residues gradually increased. The highest binding capacity of about 26%-29% was observed in clay soil under aerobic conditions after 90 d. A good balance sheet was obtained and the percentage recovery was generally between 91% and 100%. CONCLUSION: The mineralization of 2,4-D in clay soil was higher than that in clay loam soil under anaerobic conditions. Under aerobic conditions, the soil type had no influence on mineralization capacity of 2,4-D during 90 d. The soil binding increased with time whereby the extractable 14C-residues simultaneously decreased. Chromatographic analysis of the methanol extractable 14C-residues of soils revealed the presence of 2,4-D as a main product together with 2,4 dichlorophenol. PMID- 12642988 TI - Effect of dietary administration of Lathyrus sativus pulse on intestinal biochemical parameters in normal and scorbutic guinea pigs. AB - OBJECTIVE: In order to investigate that ascorbic acid deficiency is responsible for lathyrus toxicity, the effect of dietary feeding of lathyrus pulse in normal and scorbutic guinea pigs for 3 months, on intestinal biochemical parameters was undertaken. METHODS: The intestinal brush border membrane (BBM) marker and xenobiotic metabolising enzymes (XME) were assayed. RESULTS: Exposure to 80% lathyrus alone and in scorbutic conditions showed significant inhibition of alkaline phosphatase (28%-30%), sucrase (19%) and gamma-glutamyl transpeptidase (GGT) (15%-27%) enzymes, while Ca(2+)-Mg(2+)-ATPase was significantly inhibited (38%) in scorbutic plus lathyrus treated group. The phase I XME (AHH) remained unchanged while the phase II enzyme glutathione-S-transferase (GST) was significantly decreased (20%-22%) in lathyrus and scorbutic plus lathyrus treated groups. Quinone reductase (QR) activity was found to be significantly decreased in lathyrus exposed group (20%). The intestinal biomarker contents including hexose (25%-34%) and phospholipids (20%-40%) were significantly reduced in lathyrus and scorbutic plus lathyrus exposed animals, while sialic acid showed a significant decrease (28%) in scorbutic plus lathyrus treated group. However, cholesterol levels were significantly enhanced (15%-28%) in lathyrus and scorbutic plus lathyrus treated animals. CONCLUSION: The results indicate that oral feeding of lathyrus pulse to guinea pigs can alter BBM parameters as well as XME, which may result in the intestinal toxicity. Further, ascorbic acid deficiency could be one of the pre-disposing factors of lathyrus toxicity. PMID- 12642989 TI - Effect of zinc on bone metabolism in fetal mouse limb culture. AB - OBJECTIVE: To determine the effects of zinc-deficiency and zinc-excess on bone metabolism. METHODS: We developed the culture model of fetal mouse limbs (16th day) cultivated in self-made rotator with continuing flow of mixed gas for six days in vitro. The cultured limbs were examined by the techniques of 45Ca tracer and X-roentgenography. RESULTS: The right limbs cultivated had longer bone length, higher bone density than the left limbs uncultivated from the same embryo; and histologically, the right limbs had active bone cell differentiation, proliferation, increased bone trabecula, clearly calcified cartilage matrix, and osteogenic tissue. Compared with the control group, the zinc-deficient group and zinc-excess (Zn2+ 120 mumol/L) group contained less osteocalcin (BGP) and 45Ca content, and lower AKP activity; whereas zinc-normal (Zn2+ 45 mumol/L and Zn2+ 70 mumol/L) groups contained more BGP and 45Ca contents, and higher AKP (alkaline phosphatase) activity. CONCLUSION: Both zinc-deficiency and zinc-excess can alter bone growth and normal metabolism. The results indicate that the culture model of fetal mouse limbs (16th day) in vitro can be used as a research model of bone growth and development. PMID- 12642990 TI - Lead can inhibit NMDA-, K(+)-, QA/KA-induced increases in intracellular free Ca2+ in cultured rat hippocampal neurons. AB - OBJECTIVE: To examine the effects of Pb2+ on N-methyl-D-aspartate (NMDA)-, K(+)- and quisqualate(QA)/kainite(KA)-induced increases in intracellular free calcium concentration ([Ca2+]i) in cultured fetal rat hippocampal neurons in order to explain the cognitive and learning deficits produced by this heavy metal. METHODS: Laser scanning confocal microscopy was used. RESULTS: The results clearly demonstrated that adding Pb2+ before or after NMDA/glycine stimulation selectively inhibited the stimulated increases in [Ca2+]i in a concentration dependent manner. In contrast, Pb2+ treatment did not markedly affect increases in [Ca2+]i induced by an admixture of QA and KA. The minimal inhibitory effect of Pb2+ occurred at 1 mumol/L, and more than seventy percent abolition of the NMDA stimulated increase in [Ca2+]i was observed at 100 mumol/L Pb2+. Evaluation of Pb(2+)-induced increase in [Ca2+]i response to elevating extracellular concentrations of NMDA, glycine or calcium revealed that Pb2+ was a noncompetitive antagonist of both NMDA and glycine, and a competitive antagonist of Ca2+ at NMDA receptor channels. In addition, Pb2+ inhibited depolarization evoked increases in [Ca2+]i mediated by K+ stimulation (30 mumol/L), indicating that Pb2+ also depressed the voltage-dependent calcium channels. Also, the results showed that Pb2+ appeared to be able to elevate the resting levels of [Ca2+]i in cultured neurons, implying a reason for Pb(2+)-enhanced spontaneous release of several neurotransmitters reported in several previous studies. CONCLUSION: Lead can inhibit NMDA-, K(+)-, QA/KA-induced increases in intracellular [Ca2+]i in cultured hippocampal neurons. PMID- 12642991 TI - Evaluation of the effect of interventions for the female drug abusers. AB - OBJECTIVE: To investigate an effective method to facilitate the physical and mental recovery of drug abusers in detoxification restoration period. METHODS: Integrated interventions were adopted to observe the changes in the physical and mental conditions of female drug abusers who had withdrawn drugs. RESULTS: Comparing behavioral changes between the two groups before and after intervention, we found that changes of score in the intervention group were all higher than those in the control group in terms of their physical symptoms or state of anxiety. CONCLUSION: It is necessary to help drug abusers understand the harm of drug-abuse, build up self-confidence and improve EQ through interventions. It will be beneficial for the drug addicts to refrain from drug taking and regain a normal life. Our study has proved that positive results can only be obtained from integrated intervention projects. PMID- 12642992 TI - A cross-sectional study of health-related behaviors in rural eastern China. AB - OBJECTIVE: This study examined the status of health-related behaviors among rural residents and the factors influencing the practice of such behaviors. METHODS: One thousand and ninety subjects aged 15 years or over in a rural community, Anhui Province, China were surveyed. A questionnaire was used to collect information on the health knowledge, attitude and behavior of the subjects. Information on health behavior included smoking, drinking, dietary habits, regular exercises, sleeping pattern and oral health behavior. RESULTS: The prevalence of smoking and drinking in the male subjects was 46.5% and 46.9%, respectively. There was a positive significant association between smoking and drinking. Only 8.3% of all subjects ate three regular meals a day regularly. Among subjects who ate two meals a day, 89.7% did not have breakfast. Only 1.7% of subjects took part in regular exercise. About 85% of subjects slept 6 to 8 h per day. Only 38.4% of the respondents had the habit of hand washing before eating and after using the lavatory. 79.3% of the subjects brushed their teeth every day, and among them, only 10.6 percent brushed their teeth twice a day. Further analyses showed that 64.8% of subjects had 3-5 items of positive health behaviors out of 8 items and only 16.9% had six or more items. Logistical regression analyses suggested that better health behavior was affected by sex, age, years of education, income and health knowledge. CONCLUSION: The status of health behaviors among rural residents was generally poor. It is thus urgent to reinforce health education in rural communities in China. PMID- 12642993 TI - Research on sexuality: achievements and prospects. PMID- 12642994 TI - [HIV prevention: the "career" of a research subject]. PMID- 12642995 TI - [AIDS prevention: an endangered innovation]. PMID- 12642996 TI - Origins and early development of the case-control study: Part 2, The case-control study from Lane-Claypon to 1950. AB - The first modern case-control study was Janet Lane-Claypon's study of breast cancer in 1926, but the design was used only sporadically in medicine and the social sciences until 1950, when four published case-control studies linked smoking and lung cancer. These 1950 studies synthesized the essential elements of the case-control comparison, produced a conceptual shift within epidemiology, and laid the foundation for the rapid development of the case-control design in the subsequent half century. PMID- 12642997 TI - [Somatic culture and HIV protection strategies of heterosexual men]. AB - OBJECTIVES: This qualitative study examines the association between the somatic culture of heterosexual men and their choice of a protection strategy against HIV/Aids. METHODS: Verbal data was generated in 23 specific interviews with Swiss German men in the age between 25 and 65 years. The analysis was based on an integration of open coding and documentary interpretation. Contrasting and systematic comparison of the cases led to a classification into types, on the basis of which the major research question could be examined. RESULTS: The study identifies four different types of somatic cultures and demonstrates that they are highly relevant for the choice of a protection strategy against HIV/Aids. CONCLUSIONS: By introducing the concept of somatic culture as a explanatory factor, this study represents an important addition to the socio-cognitive models of protection and risk behaviour in HIV/Aids. It contributes to a theoretically broader understanding and better targeted prevention for heterosexual men. PMID- 12642998 TI - [Patterns of risk behavior and aspects of authoritarianism: results of an empirical study of 20-year-old men]. AB - OBJECTIVES: As possible explanations for risk behaviour in young men past strains, lack of resources, social individualization processes and genderspecific aspects are discussed. For this study the concept of authoritarianism was used as the model for analysis. METHODS: 4,235 recruits from the German-speaking part of Switzerland were surveyed with regard to risk behaviours in sexuality, alcohol consumption, suicide tendency, and violence. The predicting variables for non risk behaviours and for different risk behaviour patterns were identified by means of logistic regression analysis. The main predicting variables included authoritarianism, disposition to discriminate, stressful life events, and personal and social resources. RESULTS: The results show that the different aspects of authoritarianism not only influence prejudice and aggression towards dissenters as has been shown in other studies, but risk behaviours in general. Between the aspects of authoritarianism and risk behaviour patterns positive as well as negative correlations were found. CONCLUSION: We conclude that there are different types of authoritarian men who differ with regard to risk behaviours. PMID- 12642999 TI - Age and gender in the management of HIV-relevant sexual risks: theoretical background and first results of a population survey in the German speaking part of Switzerland. AB - OBJECTIVES: Health related lifestyles can be seen as resources for preventive behaviour in HIV-relevant situations. A concept was developed to analyse the management of HIV-relevant sexual risks and to uncover patterns of sexual lifestyles. In this paper first descriptive results will be presented focusing on gender and age. METHODS: 2,275 men and women between the ages of 19 and 65 from the German speaking part of Switzerland were interviewed in a computer-assisted telephone survey. Data were collected concerning attitudes, behaviours and resources, situational aspects and communication with regard to health and sexuality. Particularly, respondents were asked whether within the past 24 months they had had either first sexual contacts with new partners or casual sex, two forms of contacts which have special HIV relevance. RESULTS: 527 men and women reported having had first or casual sexual contacts. First results show a substantial lack of condom use among persons of both sexes in the age group older than 45. In general men showed more risk behaviour than women. CONCLUSIONS: For the first time in Switzerland, detailed data were collected from persons older than 45 years regarding sexual risk behaviour. Older, sexually active people are a vulnerable target group which has been neglected thus far in terms of research and preventive measures. PMID- 12643000 TI - [Mortality of 1880-1980 generations in Switzerland: a rereading of evolution of mortality based on longitudinal data]. AB - OBJECTIVES: Seek to explain the declining mortality rates observed in Switzerland using a different approach from conventional period analyses. METHODS: Analysis of the mortality rates for cohorts born between 1880 and 1980 in Switzerland. RESULTS: While the mortality transition was a constant feature of the 20th century, mortality receded most sharply among the 1900 to 1940 birth cohorts. Life expectancy gains were much lower for the pre-1900 cohorts, while early trends for the post-1940 cohorts point to a slowdown in the rate of mortality decline. These findings may be connected with the social and health conditions in which the different cohorts lived. CONCLUSION: In contrast to the cross-sectional analysis, the longitudinal approach shows that the decline in mortality suddenly stops with the 1940 cohort. PMID- 12643001 TI - Cardiovascular risk factors in an Iranian urban population: Tehran lipid and glucose study (phase 1). AB - OBJECTIVES: Coronary artery disease is becoming more prevalent in developing countries, particularly in urban areas. This study was conducted to determine the prevalence of cardiovascular risk factors among Tehran urban population. METHODS: The prevalence and distribution of high blood pressure, cigarette smoking, dyslipoproteinemia, diabetes mellitus, and obesity was determined in 15,005 subjects, aged three years and over, selected by cluster random sampling in Tehran urban district-13 between February 1999 to August 2001. Total energy intake, percent of energy derived from protein, carbohydrate, and fat were assessed in a subsidiary of 1,474 persons by means of two 24-hour dietary recalls. RESULTS: In adults, 78% of men and 80% of women presented at least one CVD risk factor. The percentage of adult women with two or more risk factors was significantly greater than the one for men. Prevalence of DM, hypertension, obesity, high TC, low HDL, high TGs, and smoking was 9.8, 20.4, 14.4, 19.3, 32, 5.3, and 22.3%, respectively. In children and adolescents, two or more CVD risk factors were found in 9% of boys and 7% of girls. Prevalence of hypertension, obesity, high TC, low HDL, and high TGs, was 12.7, 5.2, 5.1, 10.2, and 5%, respectively. The mean percentage values of energy intake derived from carbohydrate, protein, and fat were 57.8 +/- 6.9, 11.1 +/- 1.8, and 30.9 +/- 7.2, respectively. CONCLUSION: The prevalence of cardiovascular risk factors among Tehran urban population is high; particularly of high total cholesterol, low HDL cholesterol levels, and high waist to hip ratio. An effective strategy for life style modification is a cornerstone of a population approach to the cardiovascular risk factors. Moreover, these results will serve as a baseline for assessment of future trends in the risk factors studied. PMID- 12643002 TI - Using percentiles to summarise data instead of means and standard deviations. PMID- 12643003 TI - Phase I trial of high-dose tamoxifen in combination with cisplatin in patients with lung cancer and other advanced malignancies. AB - BACKGROUND: Tamoxifen has been reported to enhance the antitumor activity of cisplatin in preclinical models by modulation of protein kinase C signal transduction and apoptosis-related pathways. METHODS: We conducted a phase I study of high-dose oral tamoxifen in combination with intravenous cisplatin, with two objectives: 1) to determine tolerability, and 2) to determine the daily tamoxifen dose required to achieve serum levels equivalent to in vitro concentrations reported to enhance cisplatin cytotoxicity in preclinical models. Tamoxifen was administered days one through seven at escalating daily doses of 160 mg/m2 (n = 5), 200 mg/m2 (n = 6), and 250 mg/m2 (n = 4) by patient cohort, followed by cisplatin at 100 mg/m2 on day eight. Serum concentrations of tamoxifen and its hydroxylated metabolite N-desmethyltamoxifen were determined by high-performance liquid chromatography (HPLC) on day eight of the first treatment cycle in seven patients. RESULTS: Fifteen patients with advanced malignancies received treatment with tamoxifen at 160 mg/m2, 200 mg/m2, and 250 mg/m2 per cycle, respectively. Serum concentrations of tamoxifen and N-desmethyltamoxifen on day eight of the first cycle ranged from 1.75-8.22 microM (mean 4.72 microM) and 3.62-10.85 microM (mean 3.87 microM), respectively. Toxicity analysis demonstrated that grade 3/4 nonhematological toxicity occurred in 0/5 at a tamoxifen dose of 160 mg/m2, 1/6 at a tamoxifen dose of 200 mg/m2, and in 1/4 patients at the 250 mg/m2 dose level. No grade 4 hematological toxicity occurred. Classic dose-limiting toxicity was not observed; the trial was closed to further accrual after documentation that targeted tamoxifen levels (around 5 microM) were achieved with daily tamoxifen doses > or = 160 mg/m2 in combination with cisplatin. CONCLUSIONS: This regimen of high-dose tamoxifen in combination with cisplatin can be safely administered. Serum tamoxifen levels comparable to concentrations required for enhancement of cisplatin sensitivity in vitro are clinically achievable with acceptable toxicity. The level of antitumor activity in nonsmall cell lung cancer NSCLC is encouraging (partial response in 4/10 patients). Based on these data, a Phase II study of high-dose tamoxifen in combination with cisplatin in patients with metastatic NSCLC is being conducted through the Southwest Oncology Group. PMID- 12643004 TI - Phase I trial of gemcitabine and paclitaxel in advanced solid tumors. AB - PURPOSE: Gemcitabine and paclitaxel are chemotherapeutic agents with clinical antitumor activity in a broad range of malignant solid tumors. Because of preclinical synergy, unique mechanisms of action and resistance, and nonoverlapping toxicities, gemcitabine and paclitaxel combinations are attractive for testing in clinical trials. Prior weekly gemcitabine and paclitaxel regimens administered on a 28-day cycle have been limited by cumulative hematological toxicity on day 15, thus reducing the planned gemcitabine dose intensity. We therefore conducted a phase I trial of a 21-day schedule of weekly gemcitabine and paclitaxel to determine the tolerability, maximum tolerated dose (MTD), and preliminary estimates of efficacy of this regimen. PATIENTS AND METHODS: Forty one patients with advanced malignant solid tumors were accrued. Gemcitabine was given at a fixed dose of 1000 mg/m2 while paclitaxel was administered at an initial dose of 60 mg/m2, then escalated by 15 mg/m2 increments over seven dose levels to a prospectively planned maximum dose of 150 mg/m2. Both agents were infused intravenously on days one and eight every 21 days. At least three patients were enrolled per dose level. No intrapatient dose escalation was allowed. RESULTS: All patients were assessable for toxicity and 31 were assessable for response. The regimen was generally well-tolerated. Dose-limiting thrombocytopenia was observed in one patient at a paclitaxel dose of 135 mg/m2/week (dose level 6). After expansion of this dose level by 14 additional patients, no further dose-limiting toxicities were observed although one patient at dose level seven died of neutropenic sepsis after completing three cycles. There were eight partial responders for an overall response proportion of 26% (95% CI: 11, 41). Twelve patients (39%) had stable disease. CONCLUSION: This 21 day schedule of gemcitabine and paclitaxel is safe, well-tolerated, and active. The recommended phase II dose is gemcitabine 1000 mg/m2 and paclitaxel 150 mg/m2 on days one and eight every 21 days. The antitumor activity observed with this regimen warrants further investigation. PMID- 12643005 TI - Prognostic factor analysis in advanced gastric cancer patients treated with hydroxyurea, leucovorin, 5-fluorouracil, and cisplatin (HLFP regimen). AB - This study was performed to determine the prognostic factors of 102 nonresectable locally advanced or metastatic gastric cancer patients prospectively treated with a multimodulation of 5-fluorouracil (5FU), hydroxyurea, leucovorin, and cisplatin. Response rate in 85 patients with measurable disease was 62.4% (95% confidence interval 51.9% to 72.9%). A weight increase (5% or more) was observed in 47% of patients, performance status improved in 70.6%, and symptoms disappeared in 69%. Median times for progression-free survival and overall survival were eight and 11 months, respectively. Liver metastases, more than two involved sites, and increased carcinoembryonic antigen (CEA) were found to be univariate adverse prognostic factors for survival. In a multivariate analysis, only the presence of liver metastasis was found to be an independent prognostic factor. Response rate and survival in patients with gastric linitis or diffuse forms were in the same range as in patients with intestinal forms of gastric adenocarcinoma. PMID- 12643007 TI - Sialometrical and sialochemical analysis of patients with chronic graft-versus host disease--a prolonged study. AB - Patients with graft-versus-host disease (cGVHD) suffer from oral dryness and increased levels of oral infections and mucosal pathologies. The purpose of the current study was dual: 1) to investigate the salivary functional (sialometry) and compositional (sialochemistry) alterations induced by the disease during a 12 month period following the onset of the disease; and 2) to evaluate the effect of Salagen 30 mg/d on the salivary biochemical and immunological composition in cGVHD patients. Significant higher concentrations of salivary sodium (Na), magnesium (Mg), total protein (TP), albumin (Alb), epidermal growth factor (EGF), and total IgG accompanied by a concomitant increase in total IgA that did not reach significant value was observed in cGVHD patients in comparison with controls at both resting and stimulated conditions (p < 0.05) while salivary levels of potassium (K), calcium (Ca), and phosphate (P) were not altered. Two weeks of oral Salagen 30 mg/d resulted in normalization of the salivary biochemical and immunological compositional alterations in the cGVHD patients. Oral pilocarpine was able to reduce and normalize the elevated levels of Na, Mg, TP, Alb, EGF, IgG, and IgA salivary concentrations at both resting and stimulated conditions. The ability of oral pilocarpine to normalize and reverse salivary biochemical and immunological alterations induced by cGVHD is parallel to its stimulatory effect on salivary flow rates, as we previously showed. As the biochemical and immunological composition of the saliva results in its antimicrobial protective characteristics, the ability of oral pilocarpine (Salagen) to abrogate cGVHD salivary gland abnormalities may be of clinical importance. PMID- 12643006 TI - Radiation-induced mucositis: a randomized clinical trial of micronized sucralfate versus salt & soda mouthwashes. AB - Oral mucositis is one of the major toxicities caused by radiation therapy (RT) treatments to the head and neck. The clinical efficacy of sucralfate (Carafate R) mouthwash for head and neck cancer patients (HNC) is not consistent across studies. In this study, it was hypothesized that if the particles in the original sucralfate suspension were micronized (i.e., < or = 25 microns) then the coating action of the mouthwash in the oral cavity would be enhanced. The purpose of this pilot study was to compare the efficacy of micronized sucralfate (Carafate R) mouthwash and salt & soda mouthwash in terms of the severity of the mucositis, the severity of mucositis-related pain, and the time required to heal RT-induced mucositis in patients with HNC. Severe mucositis and related pain can interfere with the ingestion of food and fluids, so patients' body weights were measured as well. All patients in this randomized clinical trial carried out a systematic oral hygiene protocol called the PRO-SELF: Mouth Aware (PSMA) Program. Patients who developed RT-induced mucositis anytime during their course of RT were randomized to one of the two mouthwashes and followed to the completion of RT and at one month following RT. Two referral sites were used for the study. Repeated measures occurred with the following instruments/variables: MacDibbs Mouth Assessment and weight. Demographic, disease, and cancer treatment information was also obtained. Thirty patients successfully completed the study. The typical participant was male (70%), married/partnered (70%), White (63%), not working or retired (73%), and had an average of 14.5 years of education (SD = 3.7). T-tests and Chi-square analyses with an alpha set at 0.05 were used to compare differences between the two mouthwashes. No significant differences were found in the number of days to onset of mucositis (i.e., 16 +/- 8.4 days). When patients had their worst MacDibbs score, (i.e., the most severe mucositis), there were no significant differences between the mouthwashes as to MacDibbs score, the RT dose received, or ratings of pain (upon swallowing). Similarly, at the end of RT, no significant differences were found between mouthwashes as to MacDibbs scores or ratings of pain (upon swallowing). At the one-month follow-up assessment no significant differences were found between the mouthwashes in MacDibbs scores or pain ratings (upon swallowing). The analysis of the efficacy of the two mouthwashes revealed no significant differences in the time to heal (in days) from the RT-induced mucositis. The findings from this trial provide important clinical information regarding cost analysis of RT mucositis management. Given that there is no significant difference in efficacy between micronized sucralfate and salt & soda, use of the less costly salt & soda is prudent and cost effective. PMID- 12643008 TI - A phase II study of 13-cis retinoic acid plus interferon alpha-2a in advanced stage penile carcinoma: an Eastern Cooperative Oncology Group study (E3893). AB - PURPOSE: Combined biological therapy with 13-cis-retinoic acid (13-cRA) and interferon alpha-2a (IFN alpha-2a) was reported to be highly effective in squamous cell carcinoma of the cervix and skin. Squamous cell carcinoma of the penis is rare in the United States, accounting for less than 1/2% of all male malignancies. Because of the association of infection with human papillomavirus with both carcinomas of the cervix and penis and their shared squamous cell histology, we carried out a phase II study of 13-cRA and IFN alpha-2a in carcinoma of the penis. MATERIALS AND METHODS: Eighteen ambulatory patients with surgically unresectable, recurrent, and/or metastatic squamous cell carcinoma of the penis were treated with IFN alpha-2a, 3MU/day administered subcutaneously and 13-cRA, 1 mg/kg orally daily for at least eight weeks, unless intolerable toxicity occurred. RESULTS: One patient was ineligible; one patient withdrew prior to treatment. Among the 16 eligible, treated patients, there was one complete response. Fourteen patients had progressive disease as their only treatment effect. Two patients were unevaluable for tumor response because they had no follow-up tumor measurements. No unexpected treatment-related toxicities were found on study. The only common form of grade 3 toxicity was hypertriglyceridemia found in eight of the 17 patients (47%). No toxicities above grade 3 were observed. CONCLUSION: In contrast to its benefit in squamous cell carcinomas of the cervix and skin, the combination of 13-cRA and IFN alpha-2a has low efficacy in advanced carcinoma of the penis. PMID- 12643009 TI - Differentiating solitary pulmonary metastases in patients with extrapulmonary neoplasmas using FDG-PET. AB - The purpose of this preliminary study is to evaluate the characteristics of solitary pulmonary lesions in patients using extrapulmonary neoplasmas with 18F fluoro-2-deoxyglucose positron emission tomography (FDG-PET). Thirty-two patients with extrapulmonary neoplasmas who had solitary pulmonary lesions and were suspected for pulmonary metastasis underwent an oncological survey with FDG-PET. Standard uptake value (SUV) and the ratio of lesion-to-background (L/B ratio) were used as parameters to differentiate and characterize the solitary pulmonary lesions. Using SUV > 2.5 or L/B ratio > 3 as the cutoff to diagnose malignancy, FDG-PET correctly identified 29 true-positive cases. However, FDG-PET failed to interpret two false-positive and one false-negative cases. The accuracy of FDG PET was 91%. We concluded that FDG-PET is an accurate modality to differentiate solitary pulmonary lesions in patients with extrapulmonary neoplasmas. PMID- 12643010 TI - DNA biomarkers antecede semiquantitative anthracycline cardiomyopathy. AB - Adriamycin (ADM, or doxorubicin hydrochloride) is an effective antineoplastic agent whose use is restricted by its well-described, dose-dependent cardiotoxicity. This study measures ADM DNA adduct formation by 32P-radiolabeling DNA, enzymatically digesting radiolabeled DNA, separating the formed adducts on two-dimensional thinlayer chromatography (2D-TLC), and quantitating the adducts with autoradiography and densitometry. Thirty-six male Sprague-Dawley rats are randomized to receive ADM at varying intraperitoneal (i.p.) injection concentrations: 0.9% saline i.p. controls, 4 mg/kg ADM i.p., and 6 mg/kg ADM i.p. Myocardial and pulmonary tissues are harvested 48 hours after i.p. injection for autoradiographic and histopathologic analyses. The results indicate differences in the amount and type of adduct formation as a function of ADM concentration. Increased partial depurination of dGMP and dAMP occurs with increasing ADM concentration at equal incubation times. This depurination correlates with the emergence of new adducts HM-dUMP, 8-OH-dGMP, HM-dCMP, and Me-dCMP. The quantification of these adducts can potentially represent an early marker of ADM cardiotoxicity and thereby optimize the efficacy of individual chemotherapy regimens while minimizing adverse effects. PMID- 12643011 TI - Primary T-cell lymphoma of the breast: a case report. AB - Primary nonHodgkin's lymphoma (NHL) occurs in both nodal and extranodal sites. Lymphoma arising in mammary tissue is rare. The majority are of B-cell origin, while a few case studies of T-cell lymphomas of the breast have been reported. The clinical and histologic features of a 74-year-old female diagnosed with T cell mammary lymphoma are reported, as well as her treatment course and follow up. A literature review is included. We conclude primary low-grade T cell lymphoma of the breast can be treated with conservative surgery followed by involved field radiation therapy. PMID- 12643012 TI - Developments in the systemic therapy of pancreatic cancer. AB - Pancreatic adenocarcinoma is the fourth leading cause of cancer mortality in the United States of America. Progress in the treatment of this disease in the past several decades has been very modest. Several new agents with activity against pancreatic cancer have been identified. Of these, gemcitabine is the most promising agent when used in combination with other drugs. Pilot phase II studies combining gemcitabine with 5-flourouracil, irinotecan, docetaxel, or cisplatin show improved outcomes in objective response rates and survival that need to be confirmed in larger randomized studies. Advancement in the understanding of the molecular biology of neoplasia in recent years has helped identify several molecular targets for future new drug development in pancreatic cancer. Assessment of response to therapy of pancreatic cancer has been a difficult challenge. Functional imaging with techniques such as positron emission tomography (PET) may yield a more precise and timely objective evaluation of response to treatment. PMID- 12643013 TI - Taxanes in the treatment of advanced (stage III and IV) non-small cell lung cancer (NSCLC): recent developments. AB - Taxanes, paclitaxel, and docetaxel have become the cornerstone of both first-line and second-line chemotherapy for advanced non-small cell lung cancer (NSCLC). Recently, several pivotal phase III randomized trials have been published. These studies and phase II trials will be discussed. Additionally, studies utilizing a taxane and radiation therapy for resectable and locally advanced NSCLC will be outlined. The article will end with a discussion on newer strategies being currently explored to improve survival in advanced NSCLC. PMID- 12643014 TI - AML1 interconnected pathways of leukemogenesis. AB - The AML1 transcription factor, identified by the cloning of the translocation t(8;21) breakpoint, is one of the most frequent targets for chromosomal translocations in leukemia. Furthermore, polysomies and point mutations can also alter AML1 function. AML1, also called CBF alpha 2, PEBP alpha 2 or RUNX1, is thus implicated in a great number of acute leukemias via a variety of pathogenic mechanisms and seems to act either as an oncogene or a tumor suppressor gene. Characterization of AML1 knockout mice has shown that AML1 is necessary for normal development of all hematopoietic lineages and alterations in the overal functional level of AML1 can have a profound effect on hematopoiesis. Numerous studies have shown that AML1 plays a vital role in the regulation of expression of many genes involved in hematopoietic cell development, and the impairment of AML1 function disregulates the pathways leading to cellular proliferation and differentiation. However, heterozygous AML1 mutations alone may not be sufficient for the development of leukemia. A cumulative process of mutagenesis involving additional genetic events in functionally related molecules, may be necessary for the development of leukemia and may determine the leukemic phenotype. We review the known AML1 target genes, AML1 interacting proteins, AML1 gene alterations and their effects on AML1 function, and mutations in AML1-related genes associated with leukemia. We discuss the interconnections between all these genes in cell signaling pathways and their importance for future therapeutic developments. PMID- 12643015 TI - Cancer drug development in Europe: a selection of new agents under development at the European Drug Development Network. AB - From may 99, three european leaders in anti-cancer drug development (EORTC, Cancer Research UK, SENDO) have got together to form a network of collaborating groups: the European Drug Development Network. The member organisations have all agreed to join their efforts in developing new drugs. They have acquired a great and efficient expertise in anticancer drug development covering all aspects from drug screening to refinement of trial methodology and translational research. In this paper, the most interesting drugs under development in each of the three organisations are being described and discussed. PMID- 12643016 TI - Current therapies in ovarian cancer. PMID- 12643017 TI - Screening of cervical cancer: where do we go from here? PMID- 12643018 TI - Severe liver dysfunction after raltitrexed administration in an HCV-positive colorectal cancer patient. PMID- 12643019 TI - Retaining school children and families in community research: lessons from the Study of Children's Activity and Nutrition (SCAN). AB - Retaining school-aged study participants poses a major challenge in any longitudinal research study. Dropouts produce bias in the remaining sample and this loss may affect study findings and their interpretation. Dominant factors that influence retention in pediatric research studies include family versus individual participation, patient management strategies of study personnel, knowledge about the condition or therapy, age and gender factors, credibility within the community, monetary incentives, and altruism. Eleven years after baseline assessment, Studies of Children's Activity and Nutrition boasts a 53% retention of the original biethnic cohort in San Diego. Retention occurred partly due to a trained measurement team which completed sequential observations primarily in family homes and implemented continuous participant follow-up procedures. Approaches for increasing student retention based on carefully designed studies and adherence indicators can assist researchers seeking maximum retention of school-aged participants. PMID- 12643020 TI - Recruitment barriers and successes of the American Lung Association's Not-On Tobacco Program. AB - This paper explores recruitment barriers and successes with research involving Not-On-Tobacco, the American Lung Association's (ALA) teen smoking cessation program. Forty-six program facilitators across four N-O-T studies completed a questionnaire to assess recruitment methods used, effectiveness of chosen methods, and recruitment barriers. Facilitators reported the most effective recruitment methods were "one-on-one conversation with students" (53.3%) and interpersonal contact where students received lollipops and information about N-O T (33.3%; "lick-the-habit table"). The most frequently reported barriers to recruitment were "students not interested" (60.9%) and "active parental consent" (28.3%). The greatest barrier to obtaining active parental consent, as reported by facilitators, was "students did not want to tell parents they smoked" (78.3%). Findings suggest that recruitment presented a challenge to N-O-T research partners, including investigators, ALA staff, and program facilitators. However, recruitment was effective when active recruitment techniques such as maximized interpersonal contact involving one-on-one conversation were used. PMID- 12643021 TI - A meta-evaluation of 11 school-based smoking prevention programs. AB - Eleven school-based smoking prevention programs were subjected to a meta evaluation. Criteria for the meta-evaluation included: 1) adequacy of the research design, 2) evidence of reliability, 3) evidence of validity, 4) appropriate statistical analyses and interpretations, 5) reporting of effect sizes or practical significance, 6) accounting for attrition, and 7) tracking of fidelity to the program. A three-point rating scale was used ranging from 0-2. Criteria with the best ratings were research design and statistical analysis. The lowest ratings occurred for reliability and validity. The remainder of the criteria ranged between 1 and 2 with minor factors accounting for the difference in ratings. Recommendations include increasing the number of evaluations that included tests of reliability and validity and calculated effect size estimates. PMID- 12643022 TI - Long-term influence of sexual norms and attitudes on timing of sexual initiation among urban minority youth. AB - This study assessed whether sexual norms and attitudes expressed during early adolescence by minority youth from economically disadvantaged urban areas produce a sustained influence on the timing of sexual initiation. African American and Latino youth attending three middle schools were enrolled in the Reach for Health study in seventh grade and followed from an average age of 12.2 to 16.1 years. Some 849 respondents answered the question, "Have you ever had sexual intercourse" at four time points: fall seventh, spring seventh, spring eighth, and spring 10th grade. Culturally tailored scales assessed sex norms and outcome expectancies, sexual responsibility, and refusal attitudes at fall seventh grade. Influence of these norms and attitudes in early adolescence on timing of first reported sexual intercourse was examined using ANOVA controlling for gender. At fall seventh grade, 30.7% of boys and 7.7% of girls reported sexual intercourse; by spring 10th grade, the figures were 74.8% and 56.4%, respectively. Those reporting greater peer involvement in sex and more positive sex outcome expectancies were more likely to have initiated sex by fall seventh grade. Through 10th grade, the higher the scores on peer norms (f = 41.08, p < .0001) and outcome expectancies (f = 5.87, p = .002) at entry into seventh grade, the earlier the timing of initiation. Higher scores on sex responsibility at baseline were associated with delayed sexual intercourse (f = 7.36, p < .001), as are refusal attitudes (f = 15.62, p < .0001). Despite significant gender differences in timing of initiation and mean scale scores, these relationships were similar for males and females. Findings suggest the importance of addressing sexual norms and attitudes of minority youth in interventions to delay early sexual initiation in urban environments where this risk is high. Given their sustained influence on timing of sexual initiation, such interventions must begin prior to middle school and continue through mid-adolescence, years when early sexual experience can lead to negative health and social outcomes. PMID- 12643023 TI - Evaluation of a child abuse prevention curriculum for third-grade students: assessment of knowledge and efficacy expectations. AB - Researchers investigated the effects of a child abuse prevention program on third grade students' knowledge of child abuse and efficacy expectations regarding abusive situations. Three hundred forty-one students completed the 26-item pretest and posttest questionnaires (n = 166 for the intervention group, n = 175 for the control group). Results indicated a statistically significant increase in child abuse knowledge from pretest to posttest with the intervention group, with no change in knowledge for the control group. However, no significant increases occurred in efficacy expectations regarding abusive situations from pretest to posttest for either the intervention or control groups. Results from this study can be used to improve child abuse prevention programs through addressing program components that may affect factors other than student knowledge of abuse. PMID- 12643024 TI - Child nutrition programs in Canada and the United States: comparisons and contrasts. PMID- 12643025 TI - Perspectives of the observer and the observed. PMID- 12643026 TI - Hong Kong Chinese family caregiving: cultural categories of bodily order and the location of self. AB - This ethnographic study entailed interviews and observations over 2 years involving 35 Hong Kong Chinese families caring for elderly coresidential members. Bodily disorder was a starting point from which to locate identifiably Chinese attempts to restore disrupted family relations. Themes included bodies out of order--challenging Confucian-informed models of family obligation; controlling bodily excretions; the body in decay; and the body as the location of self. The management of bodily order by Chinese caregivers provides a cultural category in which personal experiences are ordered as public matters within specific local contexts. Caregivers dealt with both the demands of bodily needs and the imperative to maintain a social relationship. The establishment of bodily and self-order reflects a Chinese social system whereby moral duties are located primarily in kinship. The author summarizes culturally specific ways in which health professionals working with Chinese families involved in long-term care can target assessment and interventions before the onset of physical and emotional deterioration. PMID- 12643027 TI - Experiencing chronic illness: cocreating new understandings. AB - This study was conducted as part of a research course in which new partnerships with area citizens and community-driven programs of research were developed. Working together, the teachers, students, and citizens were able to document their practical knowledge through conducting a study of the lived experiences of chronic illness using Heideggerian hermeneutical phenomenology. The pattern, Experiencing Chronic Illness: Cocreating New Understanding, and three themes emerged during the analysis of the data (a) focusing on functional status doesn't adequately account for the experience of chronic illness, (b) decentering the focus on the treatment of symptoms makes way for equally important discussions of meaning making in the context of chronic illness, and (c) the objectified language of healthcare covers over how chronic illness is experienced. PMID- 12643028 TI - Turning points and protective processes in the lives of people with chronic disabilities. AB - In this qualitative study, the authors examined the nature of resilience in people with chronic disabilities. Fifteen people with disabilities identified the factors that helped or hindered them at major turning points, and the triggers and resolutions to these turning points. Turning points were emotionally compelling experiences and realizations that involved meaning acquired through the routes of belonging, doing, or understanding the self or the world. The major protective factors were social support, traits such as perseverance and determination, and spiritual beliefs. Three new protective processes were identified: replacing a loss with a gain (transcending), recognizing new things about oneself (self-understanding), and making decisions about relinquishing something in life (accommodating). These protective factors, processes, and ways in which people with disabilities draw sense and meaning in life have important implications for service delivery. PMID- 12643029 TI - Community care for people with disability: blurring boundaries between formal and informal caregivers. AB - Health care for individuals with disability is increasingly shifting from institutional settings to the "community," with assistance by those in the formal sector. In this article, the authors examine 5 case studies illustrating interactions and relationships between people with disability and their caregivers, using qualitative data collected as part of a community study of disability conducted in a medium-sized city in the northeastern United States. Employing the task specificity framework, they explore the implications of using either formal care providers to fill needs that are more typically met by family and friends or family caregivers to provide care that is best provided by the formal sector. Although our narratives illustrate the negative implications of mismatched care substitution, we conclude that the framework is less applicable to emerging systems of community care. PMID- 12643030 TI - Embracing uncertainty: an exploration of the experiences of childhood cancer survivors. AB - Uncertainty is a central theme in the cancer survivorship and coping literature. Existing research on cancer-related uncertainty has primarily documented the adverse effects of existential uncertainty on survivors' well-being and located uncertainty within the stress and coping paradigm. Using in-depth qualitative interviews, the authors explores the paradoxical meanings and impact of uncertainty in the lives of long-term survivors of childhood cancer. The findings suggest that although uncertainty can be a source of distress, it can also be a catalyst for growth, a deepened appreciation for life, greater awareness of life purpose, development of confidence and resilience, and optimism. The results suggest that uncertainty is a dialectic phenomenon located within larger contexts of both stress and coping, and psychospiritual growth. PMID- 12643031 TI - Survivorship as craft and conviction: reflections on research in progress. AB - The choice of some illness survivors to engage in service to other ill people, and thus to focus their lives on illness longer than their own treatment requires, is not self-evident. The authors describes alternative cultural narratives of survivorship, proposes an understanding of survivorship as a form of craft, and considers the societal mortality that is reflected in survivor's accounts of their service work. PMID- 12643032 TI - The body in health and illness. AB - In this keynote address, the author discusses perception of the body in the context of chronic illness compared with that of health. She describes changes that occur in illness with respect to time, space, morality, aesthetics, morality, technology, information, and interpersonal relationships using examples from her research, and explores the construction of illness and health identities. PMID- 12643033 TI - Accomplishing care at home for people with dementia: using observational methodology. AB - The authors consider the problems and possibilities presented by using unstructured observation in the home setting. The findings of the original study are described elsewhere (Briggs, Askham, Norman, & Redfern 1998; National Health Service [NHS] Executive 1998). In this article, the authors discuss process issues (e.g., gaining and maintaining access to the research setting, disengaging, what can and cannot be observed, and the validity of the data and analysis), drawing on a study of the nature of care as a form of social organization in the homes of people with dementia. They show that observation is not only possible but, when combined with conversational interviewing, essential for understanding the processes of caring and what it means to live with dementia. PMID- 12643034 TI - Evaluating health system change--using focus groups and a developing discussion paper to compile the "voices from the field". AB - The authors describe the contribution of a series of focus groups and a progressively revised discussion paper to the evaluation of health care reform in South Australia. The focus groups informed the evaluation about participants' experiences of the impact of reform on their agencies while enabling increased understanding about change in the health system. A discussion paper describing global health service trends and emerging findings from our evaluation was prepared prior to the first focus group and subsequently revised to incorporate participants' comments before being sent to the next group. The results was a paper that enhanced participants' discussion, and reflected their views. As an action research-based method, this iterative process was valuable for participants and the evaluation, and supported organizational learning. PMID- 12643035 TI - Assessing (audience) construction hazards: depiction of audience as a variable for comparison of health interventions. AB - Current literature regarding health promotion and strategic communication lacks sufficient inquiry regarding the communication assumptions underlying many efforts in that arena and the implications of those assumptions. In addressing that void, the nature of the intended audience could be useful as a variable. Depictions of intended audiences often are apparent in official talk about an intervention, such as in reports or other intervention materials. Examining the language that intervention developers use to characterize intended audience members affords insight regarding the relationships between health professionals and the individuals they seek to engage. PMID- 12643036 TI - Pharmacokinetics of intramuscularly administered biperiden in guinea pigs challenged with soman. AB - Biperiden is an anticholinergic compound that has demonstrated effectiveness for treating organophosphate-induced seizure/convulsions. The plasma levels of biperiden associated with this efficacy have not yet been defined. In this study, the pharmacokinetics and tissue distribution of biperiden after intramuscular administration of 0.5 mg/kg were conducted while monitoring pharmacodynamic (electroencephalographic) data in soman-exposed guinea pigs. Overall, 59% of the animals had seizures terminated within 30 min of the biperiden administration. The mean time to seizure termination was 15.9 min. The pharmacokinetics of biperiden after i.m. administration to guinea pigs were best described by a one compartment model with first-order absorption and elimination. The maximal plasma biperiden concentration (34.4 ng/mL) in seizure-terminated animals occurred at 26.3 min. Extensive partitioning into peripheral tissues was noted supporting the relatively large volume of distribution observed. Maximal biperiden concentrations in the cortex and brain stem were found at 30 min and were 2.3 and 1.7 times greater, respectively, than that in plasma. The time for maximal plasma concentration was found to corresponded well with the mean time to seizure termination following drug administration. PMID- 12643037 TI - Evidence from studies with intact mammalian cells that merbarone and bis(dioxopiperazine)s are topoisomerase II poisons. AB - A Chinese hamster V79 cell-based assay for detection of topoisomerase II (topo II) poisons and catalytic inhibitors has been applied to study two bis(dioxopiperazine)s (ICRF-187 and ICRF-154) and a structurally distinct but related compound, merbarone. All three compounds have been previously characterized as being catalytic inhibitors of DNA topo II based primarily on in vitro studies with purified enzymes. The present studies indicate, to the contrary, that all three compounds are very potent DNA clastogens in V79 cells, by virtue of their ability to produce micronuclei, the formation of which is strongly antagonized under conditions in which DNA topo II is rendered catalytically inactive. None of the compounds could be demonstrated to possess catalytic inhibitory activity in intact V79 cells under the conditions tested. These studies provide biological evidence that bis(dioxopiperazine)s are capable of functional topo II poisoning in intact mammalian cells. PMID- 12643038 TI - Evaluation of the developmental toxicities of ethanol, acetaldehyde, and thioacetamide using FETAX. AB - Potential mechanisms of the developmental toxicities of ethanol, acetaldehyde, and thioacetamide were evaluated using frog embryo teratogenesis assay-Xenopus (FETAX). Early X. laevis embryos were exposed to ethanol and thioacetamide in two separate definitive concentration-response tests with and without differentially induced exogenous metabolic activation systems (MAS) or selectively inhibited MAS. Two concentration-response tests were also performed with ethanol metabolites, acetaldehyde and acetic acid. The MAS was treated with 3,4-amino 1,2,4-triazole to modulate CYP2E1 activity, and heat to inactivate flavin containing monooxygenases (FMO) activity. Results from these studies suggested that thioacetamide may be bioactivated by both CYP2E1 and the FMO systems. Ethanol also appeared to be bioactivated by CYP2E1. Acetaldehyde was markedly more potent as a developmental toxicant than ethanol or acetic acid. Binary joint mixture studies conducted with ethanol and acetaldehyde indicated that the parent compound and metabolite acetaldehyde acted in a response additive manner. These results warrant the continued use of FETAX as a means of evaluating mechanisms of developmental toxicity in vitro. PMID- 12643039 TI - Toxicology and toxicokinetics of acute and subchronic administration of histamine dihydrochloride in rats. AB - Histamine dihydrochloride is currently being evaluated as an adjuvant to immunotherapy regimens in neoplastic and infectious diseases. The no-observed effect-level (NOEL), no-observable-adverse-effect-level (NOAEL), and pharmacokinetics of subcutaneously administered histamine dihydrochloride were determined via 5 and 28 day repeated dose studies in Sprague-Dawley rats. In the five day study, male rats received 0 (vehicle), 5, 30, 500, or 1000 mg/kg BID. Acute tissue damage was observed at one or more injection sites in the two highest dose groups after 24 h. At five days, animals in these groups displayed indications of pathological inflammation at the injection sites. In the 28 day study, male and female rats received 0 (vehicle), 0.5, 5, or 100 mg/kg BID. The most significant treatment-related pathological findings were signs of inflammation at the injection sites for animals in the 100 mg/kg BID group. Hematology and clinical chemistry changes in the highest dose groups in both studies were consistent with inflammation and anemia but were found to be reversible following a 14-day recovery. Plasma histamine levels were quantified from male and female animals receiving 0.5, 5, and 100 mg/kg injections on Day 1 and 28 of the twenty-eight day study. Cmax was achieved within 0.25 h and was dose-proportional. The elimination half-life and tmax were longer at the 100 mg/kg dose than the lower doses. No marked differences between genders or between Day 1 and 28 were found. Based on these findings, the NOEL and NOAEL were established at 0.5 mg/kg BID and 5 mg/kg BID, respectively. When converted to human equivalent dose, the NOAEL is 0.81 mg/kg which is 54 times the intended human dose. These studies support a wide safety margin for histamine dihydrochloride. PMID- 12643040 TI - CalcDose: a software for drug dosage conversion using metabolically active mass of animals. AB - This Visual Basic computer program has been developed for drug dosage conversions using metabolically active mass (MAM) of the animals. The two body weights (one with known dosage and the other, for which the dosage has to be calculated) and the known dosage are entered in the respective input boxes and the appropriate units are selected using the option buttons. The program displays the report in the form of both the animals' body weights and the respective dosages in milligram per kilogram body weight as well as the total actual doses in milligrams. The object oriented layout, flexible data entry and comprehensive report format render the CalcDose software a convenient and handy tool for dosage conversions. PMID- 12643041 TI - Influence of chloroquine treatment and Plasmodium falciparum malaria infection on some enzymatic and non-enzymatic antioxidant defense indices in humans. AB - BACKGROUND: It is known that malaria infection is accompanied by increased production of reactive oxygen species (ROS) and that malaria parasites are sensitive to oxidative damage. This has been proved by the efficacy of some antimalarial drugs that are known to act via generation of ROS when administered clinically or experimentally. OBJECTIVE: There is lack of information on the effect of chloroquine on the antioxidant defense systems of normal and malaria infected humans. Since chloroquine has remained the mainstay of therapeutic regimen in malaria endemic zones, the present investigation was therefore undertaken to study the status of blood antioxidant defense mechanism, and oxidative stress following chloroquine treatment in normal and plasmodium infected humans. METHODS: Ten healthy persons (5 males and 5 females) with the same age range (18-35 years) were taken as control group. Ten other individuals were treated with 25 mg/kg body with chloroquine over three days. Ten patients with malaria, not under antimalarial therapy were taken as another group, while another set of 10 patients with malaria were treated with 25 mg/kg body weight over three days. RESULTS: The activity of superoxide dismutase was increased by 23% in individuals treated with chloroquine compared to controls while the activity of the enzyme decreased by 26% in malaria patients and by 43% in malaria patients treated with chloroquine. In all the treatment groups, the activities of catalase and glutathione peroxidase were lowered (P < 0.001). Similarly the levels of vitamins A, C, and beta-carotene were decreased in the treatment groups while plasma ceruloplasmin was increased in the groups. Glutathione and cholesterol levels were decreased while malondialdehyde level was increased significantly. CONCLUSION: Chloroquine treatment mediated oxidative stress in the host and this effect was exacerbated in Plasmodium falciparum infected patients administered with the drug. PMID- 12643042 TI - Helicase mechanisms and the coupling of helicases within macromolecular machines. Part II: Integration of helicases into cellular processes. AB - In Part I of this review [Delagoutte & von Hippel, Quarterly Reviews of Biophysics (2002) 35, 431-478] we summarized what is known about the properties, mechanisms, and structures of the various helicases that catalyze the unwinding of double-stranded nucleic acids. Here, in Part II, we consider these helicases as tightly integrated (or coupled) components of the various macromolecular machines within which they operate. The biological processes that are considered explicitly include DNA replication, recombination, and nucleotide excision repair, as well as RNA transcription and splicing. We discuss the activities of the constituent helicases (and their protein partners) in the assembly (or loading) of the relevant complex onto (and into) the specific nucleic acid sites at which the actions of the helicase-containing complexes are to be initiated, the mechanisms by which the helicases (and the complexes) translocate along the nucleic acids in discharging their functions, and the reactions that are used to terminate the translocation of the helicase-containing complexes at specific sites within the nucleic acid 'substrate'. We emerge with several specific descriptions of how helicases function within the above processes of genetic expression which, we hope, can serve as paradigms for considering how helicases may also be coupled and function within other macromolecular machines. PMID- 12643043 TI - Photosystem II: the engine of life. AB - Photosystem II (PS II) is a multisubunit membrane protein complex, which uses light energy to oxidize water and reduce plastoquinone. High-resolution electron cryomicroscopy and X-ray crystallography are revealing the structure of this important molecular machine. Both approaches have contributed to our understanding of the organization of the transmembrane helices of higher plant and cyanobacterial PS II and both indicate that PS II normally functions as a dimer. However the high-resolution electron density maps derived from X-ray crystallography currently at 3.7/3.8 A, have allowed assignments to be made to the redox active cofactors involved in the light-driven water-plastoquinone oxidoreductase activity and to the chlorophyll molecules that absorb and transfer energy to the reaction centre. In particular the X-ray work has identified density that can accommodate the four manganese atoms which catalyse the water oxidation process. The Mn cluster is located at the lumenal surface of the DI protein and approximately 7 A from the redox active tyrosine residue (YZ) which acts an electron/proton transfer link to the primary oxidant P680.+. The lower resolution electron microscopy studies, however, are providing structural models of larger PS II supercomplexes that are ideal frameworks in which to incorporate the X-ray derived structures. PMID- 12643044 TI - Mechanisms of metalloenzymes studied by quantum chemical methods. AB - The study of metalloenzymes using quantum chemical methods of high accuracy is a relatively new field. During the past five years a quite good understanding has been reached concerning the methods and models to be used for these systems. For systems containing transition metals hybrid density functional methods have proven both accurate and computationally efficient. A background on these methods and the accuracy achieved in benchmark tests are given first in this review. The rest of the review describes examples of studies on different metalloenzymes. Most of these examples describe mechanisms where dioxygen is either formed, as in photosystem II, or cleaved as in many other enzymes like cytochrome c oxidase, ribonucleotide reductase, methane mono-oxygenase and tyrosinase. In the descriptions below high emphasis is put on the actual determination of the transition states, which are the key points determining the mechanisms. PMID- 12643045 TI - Rebasing the annual mid-year population estimates for England and Wales. AB - The ONS produces mid-year population estimates annually, which are based on updating from the most recent census. Therefore, whenever results become available from a census, a new base is created for the population estimates. This has implications for historic series, which need to be revised to be consistent with both the past and the most recent census. This article describes the methodology that will be used for this rebasing of the mid-year population estimates following the availability of results from the 2001 Census. Census results also provide a unique opportunity to assess the accuracy of the population estimates that are based on the previous census and this article also describes the approach that will be taken to the assessment of accuracy. PMID- 12643046 TI - Variant population projections for the United Kingdom and its constituent countries. AB - Interest has been growing steadily in understanding the impact of the inherent uncertainty in the national population projections. As a result, the Government Acturay's Department has recently produced the most extensive set of official variant projections ever published in the United Kingdom. These include 'standard' variants based on alternative high and low assumptions for future fertility, life expectancy and net migration and also a number of 'special case scenarios.' All of these variants have been produced at both UK and individual country level. This article describes the full range of variant projections available from the 2000-based national projections and summaries some of the key results for both the UK and the individual countries. PMID- 12643047 TI - Local marriage markets in Great Britain: how diverse? AB - Estimates are made of the number of potential marriage partners available for unmarried men and women, by age, in Great Britain in 1991 and how this varies across local districts. The preferences of men and women in relation to partner ages are taken into account in the estimates. Average partner supply declines by age for women and increases with age for men. Marriage markets differ between local areas but the differentiation is not as substantial as in many other aspects of local demography and is a good deal less than the variation that occurs through time. Young women and older men have advantageous marriage markets almost everywhere while young men and older women are at a disadvantage in almost all areas. PMID- 12643048 TI - The effect of changes in timing of childbearing on measuring fertility in England and Wales. AB - Changes in the ages at which women give birth to their children mean that fertility measured at a particular point in time (period) may not be a good representation of the ultimate fertility of those women. The common measure of period fertility is the total fertility rate, which in 2001 has fallen to the lowest level since records began in England and Wales. This article presents various methods that have been proposed to adjust period fertility data to take account of changes in the timing of childbearing, applied to England and Wales data. The article concludes that while these adjustment methods provide useful insights, for example, that the total fertility rate has underestimated period quantum fertility since the 1970s, the measures produced are difficult to interpret. This is in part because the concept they are trying to measure, period quantum is itself imprecise. The adjustments do not necessarily provide a reliable indicator of underlying cohort fertility. PMID- 12643049 TI - One-parent families--and the dependent children living in them--in Great Britain. AB - This article provides updated final estimates of the number of one-parent families, and of the number of dependent children living in them, from 1995 to 1997, inclusive, together with provisional estimates for 1998 to 2000. The existing methodology has been extended so that additional alternative estimates have been generated from which to assess the "best estimates". The number of one parent families in Great Britain is provisionally estimated at 1.75 million in 2000, and the number of deppendent children living in those families as 2.9 million. PMID- 12643050 TI - [Information as a mode of action]. PMID- 12643051 TI - [Palmar sweating and facial blushing]. PMID- 12643052 TI - [Complaints and quality of neurosurgery]. PMID- 12643053 TI - [Claims to the Norwegian Patient Compensation System after neurosurgical treatment]. PMID- 12643054 TI - [Malignant intracranial tumors--repeated resection and adjuvant therapy]. PMID- 12643055 TI - [Intracranial tumors in children]. PMID- 12643056 TI - [Intracranial tumors in adults]. PMID- 12643057 TI - [Thoracoscopic sympathectomy--surgery for palmar hyperhidrosis and facial blushing]. PMID- 12643058 TI - [Transfacial neurosurgery in skull base tumors]. PMID- 12643059 TI - [Complaints in connection with total hip replacement in hip osteoarthritis]. PMID- 12643060 TI - [Psychotherapy supervisors in Norway]. PMID- 12643062 TI - [Drug therapy]. PMID- 12643061 TI - [Poisonings with benzodiazepine-like drugs]. PMID- 12643063 TI - [Ethical challenges in introducing new medical technology]. PMID- 12643064 TI - [Patient satisfaction with care during hospitalization abroad]. PMID- 12643065 TI - [Is the concept of disease needed in health services?]. PMID- 12643066 TI - [New forms of cooperation in international health work]. PMID- 12643067 TI - [A prescription for a healthier Norway--what is physician's role?]. PMID- 12643068 TI - [Treatment of hypothyroidism]. PMID- 12643069 TI - [Alcohol abuse and relatives]. PMID- 12643070 TI - [Secondary prevention after ischemic stroke]. AB - This paper reviews secondary prevention of stroke by the therapy of vascular risk factors, anticoagulation, surgical and endovascular procedures. Two recently published studies, the PROtection aGainst REcurrent Stroke Study (PROGRESS) and the Heart Protection Study (HPS) demonstrated for the first time the efficacy of antihypertensive and lipid lowering by statins in stroke secondary prevention. PROGRESS has shown that the combination of perindoprile and indapamide reduced the occurrence of ischemic and hemorrhagic strokes in hyper- and normotensive patients by 40%, whereas HPS demonstrated a 20% reduction of ischemic strokes in cases with normal or elevated serum cholesterol. Symptomatic carotid stenoses with a distal degree of > or = 70% should undergo endarterectomy; in the presence of a distal degree of stenosis of 50-69% an individual treatment decision is performed; carotid surgery is not indicated in < 50% stenoses. Patients with a cardiac source of embolism (except those with cardiac myxoma or bacterial endocarditis) should be anticoagulated with a target INR of 2.5 (range 2-3). Patients who have no indication for vascular surgery or anticoagulation will be treated with platelet inhibitors. Aspirin 100 mg/d or the combination aspirin dipyridamole are the treatment of choice. If cerebral ischemia reoccurs with aspirin or in case of aspirin intolerance clopidogrel will be administered. Patients with cerebral ischemia occurring while they are treated with clopidogrel may receive an oral anticoagulation with a target INR of 2.0 (range 1.5-2.5). PMID- 12643072 TI - [Signs of extensive myocardial infarct in the ECG of a 35-year-old smoker?]. PMID- 12643071 TI - [Erectile dysfunction: reasonable diagnostics and treatment in general practice]. AB - The availability of efficacious oral drugs has radically changed the diagnostic and therapeutic approach to erectile dysfunction. Complicated examinations as well as invasive treatment options have been widely abandoned. Instead the management of impotent men has become much more pragmatic and focused on the symptom. Consequently only a minority of impotent men needs to be referred to an urologist, which makes the therapy of erectile dysfunction increasingly attractive for general practitioners. However, successful treatment first of all still needs time and a genuine interest in the field of erectile dysfunction. In this article a reasonable diagnostic evaluation of impotent patients in general practice is described. Furthermore indication and use of little or non-invasive therapies are discussed. PMID- 12643073 TI - [Unclear exanthema and recurrent diarrhea in an airline pilot]. PMID- 12643074 TI - Voluntary discontinuance of seven years successful insulin prophylaxis produced shortly overt diabetes in a first-degree relative of a diabetic child at high risk for type 1 diabetes. AB - This is the story of a nondiabetic first-degree relative of a child with Type 1 Diabetes who was screened for Type 1 diabetes and was found to be at high risk being positive for genetic, immunologic and metabolic markers. He accepted to be treated with low-dose subcutaneous insulin and for 7 years he has been living in subclinical prediabetes state. At the beginning of eighth year, he voluntary discontinued the preventive insulin therapy and after 18 months clinically overt diabetes appeared. PMID- 12643075 TI - Lactoferrin: mechanism of action, clinical significance and therapeutic relevance. AB - Lactoferrin is an iron binding protein involved in a large spectrum of biological actions including antimicrobial actions. Lactoferrin plays a central role in ferrokinetics: it binds free iron with great affinity limiting the amount of ions available for microorganism's metabolism. Its role in the host defence mechanisms consists in bacteriostatic and bactericidal effects; moreover it inhibits the proliferation of other microbes such as fungi and viruses. Lactoferrin is also involved in the modulation of immune system and recent studies indicate that lactoferrin directly modulates both production and function of neutrophils and monocytes. PMID- 12643076 TI - Proximal forearm fractures: our clinical experience. AB - Forearm traumas are frequent and sometimes complex; soft tissue damages associated with bone lesions could be underestimated and so compromise the final result, in spite of a good reduction and osteosynthesis. Articular and iuxta articular proximal radius and ulna fractures, which can be associated to radio humeral dislocations in Monteggia lesions, are less frequent compared to diaphyseal fractures and can present a lot of therapeutical difficulties and a high rate of complications. These complications are avoided and reduced if the treatment is quick and focused on stable osteosynthesis and anatomical reduction of the fragments of the fracture and followed, as soon as possible, by rehabilitation. The authors present the 10 years experience of Parma Orthopaedics Institute with a follow-up of at least 12 months. PMID- 12643077 TI - Metallosis due to impingement between the socket and the femoral head in a total hip prosthesis. A case report. AB - Metallosis is usually defined as aseptic fibrosis, local necrosis, or loosening of a device secondary to metallic corrosion and release of wear debris. It is an occasional but characteristic clinical finding in patients who have a metal-on metal design of total hip replacement, or when metal surfaces contact after a failure or erosion of the polyethylene component. We report the case of a patient who had severe metallosis caused by wear of the femoral head with the titanium socket, after the complete erosion and destruction of the polyethylene cup, with failure of the accetabular component. Breakage of the acetabular cup constitutes one of the possible long-term complications occurring as a result of a total hip replacement. This extremely rare complication can occur at any time postoperatively and may not be associated with symptoms or other problems for a long time. PMID- 12643078 TI - Sharing the responsibility for use of seclusion and restraint. PMID- 12643079 TI - Effects of suicidal behavior on a psychiatric unit nursing team. AB - 1. Suicide of a psychiatric inpatient can have significant, although diverse, effects on the emotions and behavior of both individual nurses and team functioning. 2. The majority of participants in this study felt there was a need for formal debriefing or counseling for themselves and their colleagues after a critical incident, although there was less consensus about the timing and structure of such an intervention. 3. This research highlights the need for advanced training of critical incident stress management team members and for flexibility in the application of critical incident stress management techniques. PMID- 12643080 TI - Boosting strategies. Enhancing the self-esteem of individuals with catastrophic illnesses and injuries. AB - This qualitative study used grounded theory methods to determine how individuals were able to live with catastrophic illnesses and injuries. Twenty-eight participants were interviewed for stories of how they endured their circumstances. Although several strategies were used, this article describes a strategy entitled "boosting," which outlines the affected individuals' efforts to improve their self-esteem, which helped them bear their circumstances. Boosting has some relationship to social comparison theory. Implications for nurses and the importance of awareness of this communication need for patients are discussed. PMID- 12643081 TI - Mental health literacy beliefs. A comparison of psychiatric trained nurses and enrolled nurses in Singapore. AB - The views of mental health professionals may influence diagnosis and management options. This study reports nurses' views about the management of three common psychiatric disorders--schizophrenia, depression, and mania. Results demonstrate the effect of greater psychiatric education and training, received by psychiatric trained nurses, on mental health literacy. PMID- 12643082 TI - Interpersonal violence issues in the nursing classroom. AB - Interpersonal violence in the United States has been called a health issue of epidemic proportions (Rudman & Alpert, 2000). Reports of interpersonal violence appear daily in popular media, and specific cases have been sensationalized. The nursing profession traditionally has attempted to respond to major health issues through education, political activity, and direct patient care. So, what have nurse educators done to prepare nurses to address this issue in their clinical practices and communities? PMID- 12643083 TI - [The problems of the child with food allergies]. AB - In France, about 20% of schoolchildren are suffering from allergic diseases. Food allergy (FA) is steadily increasing for the last 20 years. Actually, 400,000 children are suffering from FA. Prevalence has increased this last decade. Frequency is estimated about 10% and increased twofold within 5 years. In France, the most common food allergens are: hen's egg (35%), peanut (24%), cow's milk (8%), mustard (6%), and fish (4%). Other food allergens as hazelnut, kiwi, wheat, sesame, seafood, and lupin. Cross reactivities to exist between fruits, vegetables pollen, latex and are more and more frequent. Multiple food allergies have recently been described. PMID- 12643084 TI - [Diagnosis and management of food allergy in the rural environment]. AB - Food allergy presents in two distinct forms: acute allergy with symptoms of anaphylactic type (acute urticaria, Quincke's oedema, anaphylactic shock); chronic allergy with symptoms of atopy (atopic dermatitis, asthma, rhinitis). Care in a rural environment includes the feature of distancing the care structure. PMID- 12643085 TI - [Food allergy as a total social fact: ethno-sociological approach]. AB - In the last few years, both the general public and researchers have witnessed a new phenomenon: food allergies. It is said that such allergies are the illness of the century, yet it is clear that they have existed for a very long time. This paper will seek to show that a food allergy cannot be understood with out the society in which it occurs. PMID- 12643086 TI - [The Individualized Welcome Project and its adaptation to the food allergy field]. AB - Allergy to food in childhood becomes today a real public health problem. Setting up some "Projets d'accueil Individualises" (i.e. a management project at school for each allergic patient) has allowed to protect the high risk pupils in the best way since 1999. Allergy adapted meals are proposed and emergency treatments they need are organized. In this field, net working seems to be essential more than everywhere else. PMID- 12643087 TI - [The problems of the child with food allergies]. PMID- 12643088 TI - [The problems of the child with food allergies. Health promotion activities]. PMID- 12643089 TI - [The bonds that have united the Madagascar Pasteur Institute and the National Madagascar Academy for a century]. PMID- 12643090 TI - [Tuberculosis in children less than 11 years old: primary resistance and dominant genetic variants of Mycobacterium tuberculosis in Antananarivo]. AB - Tuberculosis during childhood is often due to Mycobacterium tuberculosis primo infection. Tuberculosis is highly prevalent in Madagascar and most people are infected during childhood. Our objectives were to evaluate the primary resistance of M. tuberculosis and to determine the genotypes responsible for recent infection in the population. Thus we studied 142 isolated strains from 97 children (66 with pulmonary tuberculosis and 31 with extra-pulmonary tuberculosis) recruited in different health centers in Antananarivo from 1997 to 2000. Excepting one strain resistant to isoniazide, all strains were susceptible to the four antibiotics (streptomycin, isoniazid, ryfampicin and ethambutol). This result confirms the low rate of primary resistance reported during the two surveys in 1994-1995 and 1999-2000. 67 strains of 1997-2000 were typed with the genetic marker IS6110, 44 has been assigned to 13 clusters containing each 2 to 8 similar strains. Some IS6110 clusters have already been reported in 1994-1995. Some genotypes observed in 1994-1995 seemed to have disappeared in 1997-2000. (As the rate of the frequency of some genetic variants according to the period are more likely due to a difference in strain virulence). Since there is minimal antibiotic resistance versus M. tuberculosis in Madagascar, one can not explain the appearance or disappearance of certain variants because of drug resistance. Rather, this is due to the virulence of the various M. tuberculosis strains. PMID- 12643092 TI - [Central inter-laboratory quality control for microscopic diagnosis of tuberculosis in Madagascar in 2000]. AB - Since June 1997, a quarterly quality control of sputum smear exam for the tubercle diagnosis, depending on double reading of slides, was implemented between both central laboratories of the Mycobacteria National Reference Centre in Madagascar (mycobactoria laboratories of Institut Pasteur Madagascar [IPM] and Institut Hygiene Sociale [IHS]--Health Ministry). In 2000, four controls were done, in the course of which 240 slides were coloured by auramine, coming both from IPM and IHS, and another 80 slides from IHS were coloured by Ziehl-Neelsen. All the results were in agreement for the samples stained with auramine, while two false negatives were found for the samples stained with Ziehl-Neelsen. The maintenance of this quality control between the two laboratories is necessary to insure the reliability of their results and the controls that they make for the peripheral laboratories. PMID- 12643091 TI - [Mycobacterium tuberculosis resistance to antitubercular agents in Antananarivo in 2000]. AB - In 1991, the National Tuberculosis control Program (NTP) of Madagascar adopted the short treatment course and the Directly Observed Treatment Strategy (DOTS), according to the recommendations of the OMS/UICTMR. Development of M. tuberculosis primary resistance to the four antituberculosis drugs (streptomycin [S], rifampicine [R], isoniazid [H], ethambutol [E]) is an indicator of the NTP efficiency. We report results from a five-year survey among patients with new smear positive pulmonary tuberculosis. Acquired resistance is assessed among recurrent cases. During the first survey, carried out in 1994-1995 in four large cities, multidrug resistance (MDR) rate to the major antituberculosis drug H and R was low, 0.25% for primary MDR and 5% for acquired MDR. No primary MDR was found in Antananarivo; on the other hand, acquired resistance rate was the highest there (22%). Because of logistical reasons, the second survey (1999-2000) was only carried out in the capital, Antananarivo. Results obtained among 789 new patients with smear positive pulmonary tuberculosis and 79 recurrents cases in 9 diagnostic centres showed low primary and acquired resistance of 11.1% to any drug. Primary resistance to one drug was 10.6%, mainly due to streptomycin 8.5%. MDR rates are comparable with those observed in 1994-1995: 0.1% for primary MDR and 4% for acquired MDR. These results show that ten years after the new NTP implementation, only a few MDR strains are circulating in Antananarivo, which suggests that NTP has been effective. PMID- 12643093 TI - [Epidemiological data on the plague in Madagascar]. AB - The first case of plague was introduced in Madagascar in 1898 in the east coast by way of boat from India. In 1921, plague reach the highlands and a large epidemic over the next twenty years. Until the beginning of the 80's, only of few case were identified, notified mostly in rural setting. However gradually it has re-emerged as a public health problem. Urban plague is located in the city of Antananarivo (resurgence in 1978 after 28 years of apparent silence) and in Mahajanga port (resurgence in 1991 after 63 years of silence). The reactivation of the Plague National Control Program from 1994 will allow better surveillance. The aim of this analysis is to update the epidemiological data on human plague in Madagascar based on reported cases obtained from the Central Lab of the Pasteur Institute of Madagascar from 1980 to 2001 (16,928 suspected cases of which 3,500 are likely positives or confirmed positives). The Plague season runs from October to March on the central highlands and July to November on the north-western coast. Sex-ratio male/female is 1.3/1, and the age-group of 5 to 25 years is more affected. The case fatality rate was 40% in the beginning of the 1980's, and decreased to 20% by the end of the 1990's. The percentage of case with pulmonary plague decrease from 15% to less than 5%. However, geographical extension is demonstrated: 4 districts in 1980, 30 districts in 1999 and 21 districts in 2001. In 2002, the diffusion of a new rapid test (reagent strip) in the primary health centres (CSB) in 42 endemic districts may help to decrease the morbidity and the letality due to plague and improve its control at the national level. PMID- 12643094 TI - [New method of molecular typing in human Enteroviruses: characterization of Madagascar "untypable" strains]. AB - Enteroviruses, members of the family Picornaviridae, are responsible for a wide variety of diseases and represent a major public health hazard. Typing of non polio enterovirus (NPEV) infection is traditionally based on a serum neutralization assay. However, this method is time-consuming, labor-intensive, expensive, and may fail to identify antigenic variation. A new molecular typing involving partial sequencing of the genome has been recently developed. In this study, 46 NPEV strains were analyzed, including 37 antigenicaly "untypeable" viruses. Partial sequencing of the C-end of the viral capsid protein VP1 and pairwise identity with the prototype strains allow us to assign a serotype for all "untypeable" viruses. The results show a large number and wide variety of Coxsackieviruses A which belong to the HEV-C species and also Echoviruses and Coxsackieviruses B of the HEV-B species. This method may be useful to identify all NPEV serotypes in Madagascar and to assess the possible impact of circulating NPEV populations, as we enter the final stage of poliomyelitis eradication. PMID- 12643095 TI - [Validation of questionnaire methods to identify Schistosoma haematobium bilharziasis hyperendemic zones in Madagascar]. AB - Schistosomiasis is a major public health problem in Madagascar. The aim of the national control program is to reduce the morbidity in hyperendemic areas. A prospective study has been conducted in Morombe and Ampanihy to elaborate a simple method to identify Shistosoma haematobium hyperendemic communities. The study included 1,373 children from 5 to 15 years old in 17 primary schools. Moderate sensitivity and Negative Predictive Value, with high specificity and Positive Predictive Value of "blood in urine" and "Schistosomiasis" have been found. Those diagnosis values increase with age. The first symptom should be used in older children. PMID- 12643096 TI - [Current epidemiological situation of bilharziasis in the Antananarivo plain]. AB - In 1994, prior to the Project of Development of the Plain of Antananarivo, an epidemiological survey was conducted in primary schools to assess the level of schistosomiasis. The results of stool examination by the Kato-Katz method demonstrated a prevalence of 4.3% among 6,169 randomized schoolchildren. The most infected four villages are located close the Mamba river. In 1999, after the realization of the project, another study had been done in the same Public Primary School to evaluate the evolution of schistosomiasis endemicity. 5,222 randomized pupils aged 5 to 16 years old took part in the study. The sex ratio was 1.4/1. The global prevalence was 1.8%. High prevalence are respectively notified in Antanandrano Primary school (23%) close to the Mamba river in the Northern part of the capitol and in Ambohitsoa Primary School (16.3%) located near the Mahazoarivo lac. Thus, between 1994 and 1999, a significant overall decrease in prevalence was observed (p < 10(-1)), although a few areas still have relatively high rates. Many factors may explain this improvement, including sanitary education, urbanization and a lack of conditions needed in the development of intermediate host. In addition, a high prevalence of ascaridiasis (79%) and trichocephalosis (67%) was observed. 4.3% of schoolchildren had been infected by Taenia sp. Schistosomiasis is hypoendemic in the plain of Antananarivo. The strategy against this disease must include an IEC programme, focusing in childhood exposure. PMID- 12643097 TI - [Effect of the supernatant from mice liver cell primary culture on the in vitro growth of Plasmodium falciparum wild isolates]. AB - Cultivation of Plasmodium falciparum has been a major research success, leading to a greater understanding of the parasite. Despite the fact that several P. falciparum clones have been maintained in continuous culture in different laboratories, research in genomics and proteomics would require parasitic material produced from fresh wild isolates. We have tested the effect of the supernatant from primary culture of mice hepatocytes on in vitro growth of P. falciparum isolates. Parasitized blood samples were collected from Madagascan malarious patients naturally infected. Isolates proliferation was assessed by use of isotopic method. The asexual erythrocytic stages of P. falciparum were grown for 42 hours in RPMI 1640-based medium plus L15 medium-based supernatant from mice liver cells culture, and in standard RPMI 1640-based medium alone. The mean of parasite growth was 1.5 times greater when the standard medium was enriched with the liver cells layer supernatant at a proportion of 10% and 15% (v/v). The usefulness of P. falciparum ex-vivo culture and of the hepatocytes in vitro primary culture is discussed. PMID- 12643098 TI - [National Network study to perpetuate the surveillance of Plasmodium falciparum sensitivity to antimalarials in Madagascar]. AB - To redefine strategy and policy to cure or to prevent malaria, there is a need to get relevant and updated data on Plasmodium sp sensitivity level to antimalarial drugs. Thus, in September 1999, the Madagascan Ministry of Health and the Institut Pasteur de Madagascar (IPM) formed a network named RER for malaria resistance surveillance. To alleviate the lack of experienced medical teams within the health centres, and due to technical and logistic matters, as part of the network activities, it was decided to give a start with the in vitro studies which are carried out at IPM. In vitro sensitivity testing is done by use of the isotopic method. Results from the study done in 2001 demonstrate that the Madagascan P. falciparum isolates are susceptible to amodiaquine (n = 215), to cycloguanil (n = 56), to pyrimethamine (n = 98) and to quinine (n = 214). One isolate (1/110 i.e. 0.9%) of mefloquine-resistant phenotype is detected from the Eastern region. P. falciparum susceptibility to chloroquine is satisfactory with 95.4% (206/216) of in vitro sensitive isolates. RER arises from the partnership and collaboration between the Madagascan Ministry of Health and the IPM. The network set-up is presented. The usefulness of the in vivo approach, and the in vitro investigations (chemosusceptibility test and screening of mutations accounting for resistance to chloroquine) to monitor the emergence and the dissemination of drug-resistant parasites in Madagascar as well as in the subregion of the Indian Ocean is discussed. PMID- 12643099 TI - [Malaria study in the cyclone risk zone: entomological , diagnostic and therapeutic approach in the southeastern region of Madagascar]. AB - Madagascar is a tropical island affected by many natural disasters. The eastern coastal zone--an area of perennial malaria transmission--is regularly exposed to cyclones. Few malaria studies have been done in this area of Madagascar, and none have examined the potential relationship between malaria and natural disasters. A mobile team spent six weeks in the fields doing three lines of research: an entomological study by catching mosquitoes and determining their species: a therapeutic study of chloroquine (CQ) and sulphadoxine-pyrimethamine (SP) according to a 14 days WHO protocol and also a study of physician's diagnostic ability. Physicians were asked to make a presumptive clinical diagnosis of all febrile patients, and these results were compared to those obtained from blood smear examinations. The entomological study found three major vectors species: Anopheles gambiae, An. funestus and An. mascarensis. The therapeutic study showed that SP was 100% effective (n = 13) and only one case of CQ treatment failure was recorded (1/15). Finally the diagnostic study demonstrated that presumptive diagnosis of malaria based on the only clinical signs leads to an over-estimation of malaria frequency. Over 68% (102/149) of febrile patients were diagnosed by physicians to have malaria while only 52 (34.9%) were proven positive. Of the 47 patients diagnosed clinically as malaria-negative, 12 (25.5%) turned out to be positive. Outbreaks of malaria during or after natural disasters in Madagascar can be successfully treated with either CQ or SP, but compliance may be better with SP since it requires only one dose. Perhaps equally important in the context of natural disasters is to have the capacity to make a definitive diagnosis, and the dipsticks should be made available. PMID- 12643100 TI - [Value of HPLC and fluorescent pteridines in estimating the age of Anopheles gambiae adults]. AB - Fluorescent pteridines are photosensitive pigments of mosquito cuticle. Their quantity decreases with time during the adult life of mosquitoes. In order to test the feasibility of the reversed-phase HPLC (High Pressure Liquid Chromatography) technique, to measure pteridine fluorescence and to estimate the calendar age, reared Anopheles gambiae of known age are used. Head and thorax were studied on mosquitoes aged 0, 5, 10, 20 days after emergence. There was significant difference between male and female of 0 and 5 days of age; the difference was not significant at 10 days of age. The level of fluorescence between a female's head and thorax non separated does not differ from the sum of fluorescence level of separated head and thorax. Pteridin fluorescent of female's head and thorax does not correlate. It decreases significantly with chronological age between 0 and 5 days either for head + thorax or for head alone. Conversely, this fluorescence quantity is weak and maintained constant beyond 5 days. The use of thorax alone of the female mosquito allows the differentiation of mosquito aged less than 5 days, between 5 and 10 days and aged more than 10 days. Reversed phase HPLC technique, at least in the way we have demonstrated, does not appear sensitive enough to estimate the age of the species An. gambiae reared in an insectarium. It seems that this technique, relatively complex to manage, does not bring a substantial advantage compared to the method of Detinova, which allows the separation of nulliparous and parous females (i.e. < or = 3 days versus > 3 days). PMID- 12643101 TI - [Hemoparasites in wild birds in Madagascar]. AB - This study aims to evaluate the prevalence and density of haemoparasites in native Malagasy birds. Among the 387 birds, belonging to 43 species sampled at six localities in different bio-climatic zones of the island, 139 (35.9%) showed at least 1 hemoparasite with, by order of frequency, Plasmodium and/or Haemoproteus (19.9%), microfilariae (13.7% of 387 birds), Leucocytozoon (11.1%) and Trypanosoma (1.0%). An analysis to further elucidate these observations took into account the interaction of different environmental variables (altitude, season, site of collection) or aspects of the birds (age, weight, sex). There is evidence that some parasites preferentially infect some bird species or families. The largest male birds harboured the highest prevalences and densities of haemoparasite, regardless of species. These findings extend knowledge of bird/blood parasite relationships of Malagasy birds and provide interesting insights, especially concerning the pathogenicity of this type of parasitism and the parasite transmission by insect vectors. PMID- 12643102 TI - [Review of genus Coquilletidia (Diptera: Culicidae) in Madagascar and description of the larva of Cq. grandidieri (Blanchard, 1905)]. AB - The genus Coquilletidia includes some of the mosquitoes involved in the transmission of numerous arbovirosis. Adults are locally abundant and very aggressive for men. In Madagascar, thus genus concerns 3 species among which 2 are endemic and had not been described at larval stage this far. The authors describe a single larva of Coquilletidia collected at Ankazobe in the Middle West of Madagascar, at an altitude of 1200 meters. The geographical distribution of Coquilletidia adults collected in various bio-climatic zones in Madagascar, combined with observed morphological characters, permit us to attribute this larva to Cq. grandidieri. PMID- 12643103 TI - [Management of breast cancers diagnosed at the Pasteur Institute of Madagascar from 1995 to 2001]. AB - Breast cancer is a great problem of public health all over the world. In developed countries, breast cancer represents the most common cancer in females. Its incidence is also increasing in developing country. In Madagascar, no data is available to estimate the real incidence and prevalence rates of breast cancer. However, the data at the Institut Pasteur de Madagascar can confirm the extent of the problem even if it is not at a national scale. The authors report the results of a retrospective study from histological examination at the Laboratory of pathological anatomy of the IPM, during 7 years. Among 2,337 cases of cancer, 16% (373) were breast cancer. Most of them were a female breast cancer (356 cases). The average age is 48 years old. 30% of the tumors were more than 2 cm in size, corresponding at least to the T2 stade from the International Union Against Cancer anatomoclinical classification. The current histological type is the infiltrating ductal carcinoma (80%), about 2/3 belong to the grade 3 of the Scarff-Bloom-Richardson histopronostical classification. Early diagnosis of the cancer is difficult because of the insufficiency of the sanitary infrastructure, particularly for cervical and breast cancers. A national policy for screening must be set up in order to decrease the rate of these invasive carcinomas. In the meantime, informing women and training all the medical staff is a priority. Recording all the data in Madagascar would be desirable. PMID- 12643104 TI - Diabetes and dyslipidemia. PMID- 12643105 TI - Combination oral agent and insulin therapy for type 2 diabetes mellitus. AB - Type 2 diabetes is a chronic metabolic disorder that results from defects in both insulin secretion and insulin action. Over time, there is progressive beta-cell failure. Combination oral and insulin therapy aims at treating both the defects in underlying pathophysiology and replacing insulin once beta-cell failure has occurred. Combination therapy is routinely used in the management of diabetes to reduce hemoglobin A1c, weight gain, and the number of insulin injections a patient requires. This article examines the effects of combining various oral agents with insulin on glycemic control, hypoglycemia, weight gain, and nonglycemic benefits of these combinations. PMID- 12643106 TI - New aspects of insulin therapy in type 1 and type 2 diabetes. AB - Tight control of the blood glucose level decreases the frequency of complications of both type 1 and type 2 diabetes mellitus. Until recently, the available short, intermediate, and long-acting forms of insulin could not readily be used to achieve tight glycemic control without introducing an unacceptably high risk of hypoglycemia or demanding an impracticably rigid lifestyle. With the introduction of faster-acting insulin analogues, lispro and aspart, and a peakless long-acting insulin analogue, glargine, the goal of safe and effective tight glycemic control may now be within reach for many patients. The use of these new insulins allows the clinician and patient an expanded range of options for achieving good control of fasting and postprandial blood glucose levels. PMID- 12643108 TI - Treatments for diabetic neuropathy. AB - The management of symptomatic diabetic sensory neuropathy presents a therapeutic challenge to the practicing physician. Two approaches are outlined in this article. First, symptomatic therapies, which will not influence the natural history of painful neuropathy, are discussed. These include, in addition to the stable glycemic control, tricyclic drugs, a number of anticonvulsant and antiarrhythmic agents, and opioid-like medications. Topical therapies and nonpharmalogic approaches are also discussed. With the exception of near normoglycemia, treatments that may slow the progression of neuropathy are experimental and include aldose reductase inhibitors, antioxidants, and other agents. Finally, the approach to patients without symptoms but with sensory loss is discussed. PMID- 12643109 TI - Drugs for cardiovascular risk reduction in the diabetic patient. AB - The diabetic patient is at increased risk for cardiac events. Lowering hemoglobin A1c levels even within the normal range is associated with less cardiac risk. Oral agents for diabetes that reduce insulin resistance and its associated cardiac risk factors in addition to lowering glucose should be used. Energetic reduction of blood pressure with an emphasis on the use of angiotensin-converting enzyme inhibitors and beta blockers will further reduce cardiac risk. Reduction of low-density lipoprotein and triglyceride levels and elevation of high-density lipoprotein levels through judicious use of statins and other anti-lipid agents is essential and will lower the rate of cardiac events in the diabetic patients even more than in the nondiabetic patients. In addition, aspirin and folic acid supplementation should be utilized. Use of a multiple risk factor management strategy with these drugs will lower morbidity and mortality, improve quality of life, and save cost for the diabetic patient. PMID- 12643107 TI - New pharmacologic agents for diabetes. AB - New agents are being developed to address the underlying endocrinopathies and metabolic disturbances of type 2 diabetes. Stimulants of the nuclear peroxisome proliferator-activated receptor gamma (PPAR gamma) are being identified to selectively improve insulin actions, and dual agonists of PPAR gamma and PPAR alpha are being evaluated for enhanced control of hyperglycemia and dyslipidemia. Novel activators of insulin receptor phosphorylation and inhibitors of receptor dephosphorylation are offering encouraging leads for new agents. Analogues of glucagon-like peptide-1 that increase glucose-induced insulin secretion may additionally increase beta-cell neogenesis from progenitor duct cells. The amylin analogue pramlintide, which suppresses glucagon secretion and reduces weight, is advancing in clinical trial. Direct stimulants of glucose utilization and partial inhibitors of gluconeogenesis are providing useful new drug templates. Thus, new pharmacologic approaches are emerging to treat the multiple lesions of type 2 diabetes. PMID- 12643110 TI - Management of the insulin resistance syndrome. AB - The insulin resistance syndrome (IRS) is a common disorder, which has important clinical implications. It is a cluster of cardiovascular risk factors that include obesity, hypertension, dyslipidemia, glucose intolerance, and type 2 diabetes mellitus. Lifestyle modifications and insulin sensitizers are among the several therapeutic strategies available for the treatment of the IRS. Optimal treatment will not only improve glycemic control, but may also significantly lower cardiovascular disease. PMID- 12643112 TI - Dietary fats and diabetes mellitus: is there a good fat? AB - As knowledge of the fatty acid functions has increased, so has the complexity of making dietary fat recommendations to people with type 2 diabetes. Oleic acid seems to offer a slight advantage over linoleic acid in reducing plasma glucose, insulin levels, total cholesterol, low-density lipoproteins (LDLs), and triglycerides, but may also have atherogenic properties through another mechanism. A diet containing a higher proportion of polyunsaturated fatty acids (PUFAs) may require a concomitant increase in antioxidant intake because PUFAs oxidize easily and are then converted to oxidized LDL, which is more atherogenic. In addition to raising total and LDL cholesterol, long chain saturated free fatty acids may interact with plasma glucose to increase insulin secretion. Omega-3 fatty acids decrease triglycerides and reduce the risk of fatal cardiac arrhythmias. Glycemic control does not appear to be adversely affected by omega-3 fatty acids at amounts of up to 3 g/d. PMID- 12643111 TI - Combating diabetic nephropathy with drug therapy. AB - Diabetes mellitus is the leading cause of end-stage renal disease and also increases the risk of atherosclerotic vascular disease. Hypertension amplifies both problems. Detection of microalbuminuria, a common and early manifestation of diabetic nephropathy and a marker for cardiovascular risk, permits early treatment to reduce progression of nephropathy and vascular disease in diabetes. Although optimal glycemic control is essential to reduce the risk of nephropathy, aggressive blood pressure lowering to a level of 130/80 mg Hg or below in hypertensive diabetic patients is as important as glycemic control. Initial drug therapy for nephropathy should include an angiotensin-converting enzyme inhibitor (or if contra-indicated, an angiotensin receptor blocker), as several large randomized double-blinded multicenter clinical trials have demonstrated an independent renoprotective effect with renin angiotensin system inhibition. The role of advanced glycation end products in the pathogenesis of renal and vascular disease in diabetes is becoming more clearly established. However, the use of therapeutic strategies directed at blocking their effect still awaits further investigation. A multifaceted intervention program that combines optimal glycemic control, lifestyle modification/cardiovascular prevention guidelines such as lipid control and smoking cessation, with appropriate antihypertensive therapy when indicated, will prevent or delay both the occurrence and progression of diabetic nephropathy. PMID- 12643113 TI - Obesity and sedentary lifestyle: modifiable risk factors for prevention of type 2 diabetes. AB - Over the past 25 years, the prevalence and incidence of type 2 diabetes have increased alarmingly worldwide and across all age, gender, and race/ethnic groups. The high economic costs, resulting from the numerous and severe complications of this disease, as well as the social burden associated with diabetes mellitus, have motivated the pursuit of prevention methods. Recently, clinical and observational studies have demonstrated the adverse effects of obesity and sedentary lifestyle, as well as the benefits of moderate weight loss and related behaviors, including increased levels of physical activity, in maintaining metabolic control and reducing the incidence of type 2 diabetes. The challenge remains to find ways to successfully deliver this message to the communities, and to evaluate the cost-effectiveness of the different delivery approaches. PMID- 12643116 TI - Have we got the new diagnostic criteria for diabetes and impaired glucose states right? Almost. PMID- 12643114 TI - Carbohydrate and diabetes: is the source or the amount of more importance? AB - There have been two approaches to examining the effects of carbohydrate on postprandial glycemia: 1) studies designed to determine the glycemic response of foods containing carbohydrate, regardless of the source, i.e., starch, sugar, or fiber (glycemic index [GI] of carbohydrates), and 2) studies comparing the glycemic response to equivalent amounts of starches or sugars. A number of food factors determine the glycemic response to carbohydrates: food form, digestibility, cooking, sugars, type of starch, presence of antinutrients, and second meal or lente effect. In people with diabetes, the severity of glucose intolerance and premeal glucose concentrations also influence the glycemic response. The GI attempts to classify individual foods (50-g portions) by the extent to which they raise blood glucose levels compared with a standard (reference carbohydrate), initially glucose and in later studies bread. Acute glycemic responses differ, however, when meals containing low GI foods are compared to meals containing high GI foods long term (measured by fructosamine or hemoglobin A1c), the outcomes are mixed. If there is an effect from the GI of foods on glycemia, it is modest at best. In other studies, when sucrose is substituted for a variety of starches--in meals or snacks and both acutely and for up to 6 weeks--the glycemic response is similar if the total amount of carbohydrate is similar. Therefore, the recommendation for persons with diabetes in regard to the glycemic effect of carbohydrates is that the total amount of carbohydrate in meals or snacks is more important than the source or type and is the first priority in the planning of meals or snacks. This has led to the implementation of carbohydrate counting, in which foods are listed as carbohydrate choices based on the amount and not the source of the carbohydrate. PMID- 12643115 TI - Alcohol consumption and risk of coronary heart disease among individuals with type 2 diabetes. AB - The epidemiologic evidence on the association between moderate alcohol intake and risk of coronary heart disease (CHD), biological mechanisms for this association, and potential hazards of alcohol intake in individuals with type 2 diabetes is reviewed here. Three prospective cohort studies have examined the association between alcohol consumption and risk of CHD among diabetics. The results indicated significant risk reductions, ranging from 34% to 79%, associated with light to moderate alcohol intake. Potential mechanisms include increased high density lipoprotein cholesterol, decreased coagulation, and enhanced insulin sensitivity. Alcohol intake is also associated with certain risks among diabetics. However, for moderate alcohol consumption, the benefits would likely outweigh the risks. PMID- 12643117 TI - Have we got the new diagnostic criteria for diabetes and impaired glucose states right? No. PMID- 12643118 TI - The role of PCI and CABG in the management of coronary artery disease in patients with diabetes. AB - The role of percutaneous coronary intervention (PCI) and coronary artery bypass surgery (CABG) in patients with diabetes mellitus (DM) is evolving. Data from clinical trials and observational studies are reviewed as well as current clinical practice guidelines. The importance of aggressive medical therapy to achieve recommended glycemic control targets, and management of usual risk factors in patients with coronary artery disease (CAD) cannot be overemphasized regardless of the revascularization therapy selected. Patients with type 2 diabetes are at increased risk for CAD, which is the cause of death in the majority of patients. Outcomes following PCI and CABG are worse in patients with DM compared to those without DM. This calls for randomized trials and other studies focused on patients with DM. PMID- 12643119 TI - The use of ACE inhibitors on diabetic patients without renal disease. AB - Strategies that interrupt the renin-angiotensin system, especially with angiotensin-converting enzyme (ACE) inhibition, reduce cardiovascular disease mortality and morbidity in high-risk persons such as those with the insulin resistance syndrome and diabetes mellitus. In the 1980s emphasis was placed on the renal protective effects of ACE inhibitors in patients with diabetes and proteinuria. During the past several years controlled clinical trials have demonstrated that ACE inhibition reduces cardiovascular disease (CVD) mortality and morbidity. This is especially important in patients in the United States, where 80% of excess mortality for diabetes mellitus is attributed to CVD. This article reviews the clinical trials in high-risk patients, especially those with diabetes, that shown beneficial CVD risk reduction with ACE inhibitors. PMID- 12643120 TI - Endothelial dysfunction in human diabetes. AB - In nondiabetic individuals, a poor response to an endothelium-dependent vasodilator in coronary vessels has been shown to increase the likelihood of a future cardiovascular event. Such prospective data are not as yet available in patients with type 1 or type 2 diabetes. However, consistent with the greatly increased cardiovascular risk in these patients, endothelial dysfunction has been almost universally found to characterize patients with type 2 diabetes particularly. Endothelial dysfunction frequently coexists with features of insulin resistance, such as the presence of small dense low-density lipoprotein (LDL) particles even in nondiabetic individuals. This association is independent of obesity and other causes of endothelial dysfunction, such as LDL cholesterol, hypertension, and smoking. In patients with type 1 diabetes, endothelial dysfunction has been found in approximately half of the studies. In some but not all studies, endothelial dysfunction has been especially severe in patients with poor glycemic control. Reversal or amelioration of endothelial dysfunction has been documented by many commonly used therapeutic agents such as successful insulin therapy, fibrates, and angiotensin-converting enzyme inhibitors, but also with some but not all agents that act as antioxidants. Long-term studies addressing the prognostic significance of endothelial dysfunction and its reversal are urgently needed to determine whether measurement of endothelial function could be used to identify individuals at risk better than can be done at present using classic risk factor assessment among patients with type 2 diabetes especially. PMID- 12643122 TI - The musculoskeletal complications of diabetes. AB - The musculoskeletal system can be affected by diabetes in a number of ways. Some of the musculoskeletal conditions seen in diabetics are the same as those found in the general (nondiabetic) population, whereas others are unique to diabetes. The hands, shoulders, feet, muscles, and skeleton are some of the frequently affected sites. Although there is often no "cure" for these problems, there are treatments available that can significantly improve function and quality of life for diabetics with rheumatologic problems. PMID- 12643121 TI - Glucose and insulin management in the post-MI setting. AB - Despite dramatic advances in the treatment of acute myocardial infarction (AMI) in recent years, patients with diabetes mellitus continue to experience disproportionately high morbidity and mortality. A substantial body of experimental and clinical data suggest that the ability of the heart to augment its energetic metabolism of glucose in the acute setting is critical to survival and functional recovery after AMI. Emerging evidence also suggests that chronic hyperglycemia may predispose to post-AMI ischemia and heart failure via adverse effects on coronary endothelial function and myocardial ultrastructure, energy metabolism, and gene transcription. A strong case can be made for intensive insulin-based control of glycemic level in the AMI patient with diabetes mellitus. PMID- 12643123 TI - Dermatologic conditions associated with diabetes. AB - The development of skin manifestations of diabetes affects up to two thirds of patients with both types 1 and 2 diabetes mellitus. Cutaneous manifestations of diabetes occur because of the microvascular complications of diabetes, impaired wound healing, and other yet undetermined mechanisms. This article summarizes the most frequent skin findings in patients with diabetes, and proposes treatment of some complications. PMID- 12643124 TI - Erectile dysfunction and diabetes. AB - Erectile dysfunction (ED) has been the most neglected complication of diabetes. It is a common abnormality that affects more than 20 million American men. The prevalence of ED in the general population between the ages of 40 to 70 years is 52%. In men with diabetes, it ranges from 35% to 75%, and occurs at an earlier age. There have been several advances in the understanding of the physiologic and biochemical mechanisms controlling penile erections. Improved techniques in diagnoses and treatment of impotence have given the male with diabetes some hope in the management of this prevalent and emotionally disturbing complication. PMID- 12643125 TI - Insulin resistance and polycystic ovary syndrome. AB - Insulin resistance plays a central role in the pathogenesis of the reproductive as well as metabolic aberrations that characterize the syndrome. Many women with polycystic ovary syndrome (PCOS) manifest a phenotype that overlaps substantially with the so-called "metabolic syndrome" or "syndrome X." These manifestations include obesity, glucose intolerance, hypertension, macrovascular disease, and dyslipidemia. The etiology and manifestations of insulin resistance in PCOS are reviewed in this article. PMID- 12643126 TI - The effect of in-utero undernutrition on the insulin resistance syndrome. AB - The metabolic and cardiovascular complications associated with in-utero undernutrition have been identified during the past 10 years. Reduced fetal growth is independently associated with an increased risk for the development of cardiovascular diseases, the insulin resistance syndrome and its components: hypertension, dyslipidemia, impaired glucose tolerance, and type 2 diabetes. All appear to result from the initial development of insulin resistance that seems to be a key component underlying this association. Several hypotheses have been proposed over the past 10 years to understand this unexpected association. Each of them points to either a detrimental fetal environment or genetic susceptibilities or interactions between these two components as playing a critical role in this context. The hypothesis that this association could be the consequence of genetic/environmental interactions remains at the moment the most attractive. Although the mechanism remains unclear, there is also some evidence that adipose tissue plays a role in the emergence of insulin resistance associated with in-utero undernutrition. PMID- 12643129 TI - Blood glucose and the brain in diabetes: between a rock and a hard place? PMID- 12643127 TI - Genetics of insulin resistance. AB - Insulin resistance, defined as the decreased ability of insulin to perform its biological functions, is likely to represent the primary physiologic defect underlying the insulin resistance syndrome (IRS), which includes insulin resistance/hyperinsulinemia, glucose intolerance and/or type 2 diabetes mellitus, visceral obesity, hypertension, and dyslipidemia. This constellation of traits is a leading cause of cardiovascular mortality and morbidity. Insulin sensitivity varies widely among individuals. Although environmental provocations including physical inactivity and caloric excess play an important role in the development of obesity and thus insulin resistance, epidemiologic and family studies show that there are also moderate genetic influences on the development of insulin resistance. Extreme forms of insulin resistance may be caused rarely by mutations in the genes for the insulin receptor and peroxisome proliferator-activated receptor gamma. However, the genetic basis for common more moderate forms of insulin resistance is likely to be polygenic and heterogeneous. Evidence further suggests that gene variants may have phenotypic influences on more than one IRS trait (so-called pleiotrophy), which may explain, in part, the clustering of these traits. This article reviews the evidence that insulin resistance has a genetic basis. Progress to date toward identifying specific gene variants are reviewed. Ultimately, the identification of specific gene variants that influence insulin resistance and other IRS traits will have profound influences on our understanding of the molecular and pathophysiologic basis of these disorders, from which new and more effective preventive and therapeutic interventions will be possible. PMID- 12643128 TI - The role of the immune system in the insulin resistance syndrome. AB - The complex relationship between immunity and the insulin resistance syndrome is likely mediated to a significant degree by cytokines and the inflammatory proteins they induce. Epidemiologic work has revealed associations between cytokines and clinically evident insulin resistance, and mechanistic studies have yielded insight into the induction of insulin resistance at the cellular level by cytokines such as tumor necrosis factor-alpha. Genetic polymorphisms significantly influence this relationship, and variations in cellular immunity as manifested by T-helper cell phenotype are likely to be important as well. Further elucidation of the link between immunity and insulin resistance may lead to more effective treatment, and potentially prevention, of the insulin resistance syndrome. PMID- 12643130 TI - Diagnosis, screening, and management of cystic fibrosis-related diabetes. AB - Diabetes is a common complication of cystic fibrosis (CF), occurring in approximately 40% of adult patients. It is clinically distinct from type 1 or type 2 diabetes, and requires a unique approach. Because of evidence that it is associated with increased morbidity and mortality, prompt diagnosis and aggressive management of CF-related diabetes (CFRD) is important. The Cystic Fibrosis Foundation held a consensus conference in 1998 to define the current standards for the diagnosis and care of this disease. This article reviews those recommendations, and presents a practical approach to the management of CFRD. PMID- 12643131 TI - HIV-associated lipodystrophy: description, pathogenesis, and molecular pathways. AB - HIV-infected individuals taking antiretroviral medications may experience changes in body shape and metabolism, commonly known as HIV-associated lipodystrophy (HIVLD). In vitro and in vivo research have revealed numerous effects of both protease inhibitors and nucleoside reverse transcriptase inhibitors on the function of various organs--most importantly adipose tissue, liver, and muscle. The metabolic abnormalities could result in an increased risk of cardiovascular disease in this vulnerable and relatively young population. Treatment strategies, normally successful in the general population, have generally been less effective in this group of people, in which the detrimental effects of the antiretroviral medications are ongoing. PMID- 12643132 TI - Molecular etiologies of MODY and other early-onset forms of diabetes. AB - Maturity-onset diabetes of the young (MODY) are monogenic forms of type 2 diabetes that are characterized by an early disease onset, autosomal-dominant inheritance, and defects in insulin secretion. Genetic studies have identified mutations in at least eight genes associated with different forms of MODY. The majority of the MODY subtypes are caused by mutations in transcription factors that include hepatocyte nuclear factor (HNF)-4 alpha, HNF-1 alpha, PDX-1, HNF-1 beta, and NEURO-DI/BETA-2. In addition, genetic defects in the glucokinase gene, the glucose sensor of the pancreatic beta cells, and the insulin gene also lead to impaired glucose tolerance. Biochemical and genetic studies have demonstrated that the MODY genes are functionally related and form an integrated transcriptional network that is important for many metabolic pathways. PMID- 12643133 TI - Diabetes mellitus and other metabolic disturbances induced by atypical antipsychotic agents. AB - Atypical antipsychotic agents offer significant advantages over older conventional antipsychotic agents. The reduction in antipsychotic drug-associated extrapyramidal symptoms and the potential reduced risk for tardive dyskinesia, compared to conventional drugs, are major advances in the treatment of psychotic patients. However, recent reports of hyperglycemia, new-onset diabetes mellitus, diabetic ketoacidosis, weight gain, and lipid abnormalities associated with atypical antipsychotic agents have emerged. A review of the recent literature and an approach to the evaluation of risk factors to aide in the safe use of atypical antipsychotic agents in presented. PMID- 12643134 TI - Emotional and quality-of-life aspects of diabetes management. AB - Patients with diabetes commonly feel overwhelmed, frustrated, or "burned out" by the daily hassles of disease management and by the unending, often burdensome self-care demands. Many report feeling angry, guilty, or frightened about the disease, and often are unmotivated to complete diabetes self-care tasks. The toll of short- and long-term complications can make the disease even more burdensome. Not surprisingly, it is a consistent finding across studies that diabetes is associated with impaired health-related quality of life (HRQOL), measured in a variety of different ways. Importantly, the relationship between HRQOL and diabetes appears to be bidirectional. Both medical and psychosocial aspects of diabetes may negatively affect HRQOL; in turn, impaired HRQOL may negatively influence diabetes self-management. Unfortunately, the concept of HRQOL in diabetes remains unclear, making precise evaluation and intervention difficult. There is growing agreement that the focus of HRQOL assessment should be on the subjective burden of symptoms, not merely on the presence of objectively identifiable problems. Proper evaluation should include both generic and diabetes specific elements of HRQOL. In this article, a comprehensive multidimensional model of HRQOL in diabetes involving six major components is introduced and described. Representative self-report questionnaires that may be valuable in assessing these components are also presented. Once the patient's most important HRQOL issues have been identified and prioritized, appropriate intervention becomes possible. The good news is that there are now a growing number of research-based interventions available for addressing almost all of the HRQOL impairments that may occur. PMID- 12643135 TI - Enhancing support via interactive technologies. AB - Despite some improvements in diabetes treatment processes, outcomes for many patients remain inadequate. Interactive health technologies (IHTs) can address many of the challenges that diabetes patients and their health systems face, and research on IHT applications for diabetes care are intensifying. This article describes recent evidence regarding the feasibility and impact of 1) clinic-based CD-ROM systems supporting behavior change; 2) automated telephone diabetes management allowing for ongoing monitoring and patient education between face-to face clinical encounters; and 3) Web-based systems focusing on a range of diabetes management goals such as enhanced emotional support for patients and improved clinician adherence to treatment guidelines. Studies in each of these areas have been somewhat encouraging. However, intervention effects have been moderate, and researchers face significant challenges in designing and implementing definitive studies. PMID- 12643136 TI - Reaching underserved populations and cultural competence in diabetes education. AB - Diabetes self-management education has gained in importance over the past decade as research has documented the benefits of such interventions in improving glucose control and reducing diabetes-related complications. Although minority populations bear a disproportionate burden of diabetes, past strategies have not addressed cultural characteristics of groups typically underrepresented in diabetes research. Recent research literature on the development of culturally competent diabetes self-management is summarized and an example of a culturally competent intervention designed for Spanish-speaking Mexican Americans is presented. Recent research is laying the foundation for future intervention development to meet the cultural needs of racial/ethnic groups. PMID- 12643137 TI - The role of peroxisome proliferator-activated receptor gamma in diabetes and obesity. AB - Peroxisome proliferator-activated receptor gamma (PPAR gamma) is a nuclear receptor, which upon activation with various natural and synthetic ligands, stimulates the transcription of genes responsible for growth and differentiation of adipocytes. Furthermore, PPAR gamma is the receptor for the insulin sensitizing thiazolidinediones, which are commonly used for the treatment of type 2 diabetes. Rare inactivating mutations of the gene encoding PPAR gamma are associated with insulin resistance type 2 diabetes, and hypertension, whereas a rare gain of function mutation causes extreme obesity. A common polymorphism (Pro12Ala) of the adipose tissue-specific gamma 2 isoform is associated with increased insulin sensitivity and decreased risk of developing type 2 diabetes. These findings indicate a central role of PPAR gamma in fat cell biology and in the pathophysiology of obesity, diabetes, and insulin resistance. PMID- 12643138 TI - Calpain 10 and genetics of type 2 diabetes. AB - Positional cloning studies conducted on a region of chromosome 2q providing evidence for linkage to type 2 diabetes implicated genetic variation at the calpain-10 gene (CAPN10) in susceptibility to type 2 diabetes. The variants identified in these studies are located in introns, rather than in coding sequence. It was proposed that the cumulative effects of a combination of variants, rather than variation at a single site, increase the risk of type 2 diabetes. Confirmation of the hypothesis that non-coding sequence variation in CAPN10 affects susceptibility to type 2 diabetes has implications for how we search for susceptibility variants and interpret results of positional cloning studies for complex disorders, and suggests a new pathway in glucose homeostasis. We review the results of follow-up studies on the CAPN10 finding, and consider the issues inherent in conclusively establishing that particular genetic variation affects a complex phenotype. PMID- 12643139 TI - Genetics of diabetes complications. AB - Long-term exposure to the hyperglycemia characteristic of diabetes patients leads to serious and frequently disabling or fatal complications. Emerging evidence suggests that genes are a significant contributor to an individual's risk of developing complications. This evidence is from evaluations of familial aggregation, differences in incidence in racial and ethnic groups, and statistical analysis of family data. Evidence to date suggests that complication genes are, distinct from the genes contributing to diabetes. Molecular geneticists have taken several approaches to identify genes contributing to complications, ranging from relatively simple analysis of specific candidate genes in small case-control comparisons to systematic evaluations of the human genome using genome scans and linkage analysis in large collections of families. Results suggest that genetic contributions to diabetes complications are diverse and complex in nature, presenting a significant challenge to researchers. Diabetes-affected families are frequently enriched for complications such as cardiovascular disease or nephropathy. In addition to their value in the study of diabetes complications, such families may be valuable resources for understanding cardiovascular disease and nephropathy in the nondiabetic population also. PMID- 12643140 TI - Medical management of obesity: a clinical imperative? PMID- 12643141 TI - Prediabetes mellitus and its links to atherosclerosis. AB - Type 2 diabetes mellitus is a chronic insidious process contributing to an overwhelming amount of morbidity and mortality, much of which is related to atherosclerosis, which often accompanies and complicates the natural history of diabetes. Although considerable attention has focused on new insights into diabetic mechanisms and emerging treatments, perhaps one of the greatest opportunities for decreasing the toll of diabetes and its late-stage complications may be intervening earlier in the disease process. Such notions redirect attention toward the concept of prediabetes mellitus as a potentially discrete syndrome, which may be identifiable in clinical practice. This article seeks to offer support for this hypothesis by considering the data surrounding prediabetes mellitus, with particular attention on the mechanistic and clinical links to atherosclerosis. PMID- 12643142 TI - Diagnosis of CAD in patients with diabetes: who to evaluate. AB - Effective diagnosis and treatment of coronary artery disease (CAD) are key to the management of patients with diabetes. Although the use of specialized cardiac testing for CAD screening in asymptomatic patients varies widely and is the source of current controversy, evidence is emerging on the prevalence and predictors of asymptomatic ischemia in diabetic patients. Accurate diagnosis and risk stratification are essential in symptomatic patients with known or suspected CAD. Noninvasive cardiac testing has an important role in these patients, although evaluation for revascularization with cardiac catheterization is warranted in high-risk circumstances. This article reviews recent information that may help guide the clinician in the appropriate use of cardiac testing in diabetic patients. PMID- 12643143 TI - Arterial calcification in diabetes. AB - Diabetes is associated with an increased prevalence of atherosclerotic vascular disease and cardiovascular mortality. In diabetic patients, medial calcification appears to be a strong independent predictor of cardiovascular mortality, it occurs particularly in those with neuropathy. Recent evidence suggests that medial calcification in diabetes is an active, cell-mediated process, similar to that observed in patients with end-stage renal disease (ESRD), in which vascular smooth muscle cells (VSMCs) express a number of bone matrix proteins that act to either facilitate or regulate the calcification process. Several bone-associated proteins (e.g., osteopontin, bone sialoprotein, alkaline phosphatase, type 1 collagen, osteocalcin) have been demonstrated in histologic sections of vessels obtained from patients with diabetes or ESRD. In in vitro experiments, high glucose induced cell proliferation and expression of osteopontin in cultured VSMCs. Hypoxia had additive effects of hyperglycemia on VSMCs. In addition, uremic serum upregulates osteoblast transcription factor Cbfa 1 and osteopontin expression in cultured VSMCs. The pathogenesis of vascular calcification in diabetes is not completely understood, although high glucose and other potential factors may play an important role by transforming VSMCs into osteoblast-like cells. Further understanding of the mechanism by which diabetes induces this complication is needed to design effective therapeutic strategies to intervene with this process. PMID- 12643144 TI - Osteoporosis and diabetes. AB - Osteoporosis is the most prevalent metabolic bone disease in the United States. Although the disease has historically been reported mostly in white women, it can affect individuals of both sexes and all ethnic groups. The presence of osteoporosis related to diabetes is not well acknowledged and the impact of osteoporosis in a diabetic patient is often not considered. Routine screening or initiation of preventive medications for osteoporosis in all patients with type 1 or type 2 diabetes is not recommended at the present time. However, in all patients with diabetes, besides optimal glycemic control, general recommendations regarding adequate dietary calcium intake, regular exercise, and avoidance of other potential risk factors such as smoking should be given. In patients who have positive risk factors for osteoporosis, or in those who present with fractures, evaluation of bone density should be done and respective preventive or therapeutic interventions should be applied. PMID- 12643145 TI - Gastrointestinal disturbances in diabetes. AB - Gastrointestinal disorders are common in patients with diabetes mellitus. As many as 75% of patients visiting diabetes clinics will report significant gastrointestinal (GI) symptoms. The symptom complex experienced may vary widely. Many patients go undiagnosed and undertreated. Patients with a history of retinopathy, nephropathy, or neuropathy should be presumed to have GI abnormalities until proven otherwise. The workup should start with a thorough patient history and appropriate laboratory, radiographic, and GI testing. In addition to pharmacologic therapy, glycemic control and dietary manipulation play an important role in managing GI disorders in people with diabetes. PMID- 12643146 TI - Psychosocial aspects of diabetes with an emphasis on depression. AB - Any chronic disease is associated with an increased prevalence of mood disorders and depression. Diabetes is unique in that it places the burdens of invasive blood glucose monitoring, regimentation of diet and exercise, and often multiple daily insulin injections into the hands of the individual. Therefore, it is not surprising that depression may be three times more prevalent in the diabetic population when compared with nondiabetic individuals. Depressed patients with diabetes have been shown to have poorer glycemic control and a higher incidence of microvascular and macrovascular complications. Treatment of the depression associated with diabetes with either pharmacologic therapy or behavioral intervention has been shown to result in improved glycemic control and quality of life. Specific side effects of commonly used antidepressant medications, which have particular bearing on the diabetic state, are discussed. Future research into the causal relationship of depression to the onset on diabetes, longitudinal studies looking at depression and the rate of occurrence of diabetic complications, and the impact of the early development of healthy coping mechanisms and the treatment of depression on the natural history of diabetes are needed. PMID- 12643147 TI - Mastopathy and diabetes. AB - Sclerosing lymphocytic lobulitis or "diabetic mastopathy" is strongly associated with type 1 diabetes but may occur occasionally in its absence. It is characterized by keloid-type fibrosis, lymphocytic lobulitis, and perivasculitis and epithelioid cells. Infiltrating cells are predominantly B lymphocytes. It may be associated with retinopathy and neuropathy, but more research is needed to verify this association. The etiopathogenesis is unknown and the disorder does not seem to predispose to breast carcinoma or lymphoma. PMID- 12643148 TI - Lipid abnormalities in the metabolic syndrome. AB - The metabolic syndrome is the constellation of adverse metabolic and clinical effects of insulin resistance. Its high and increasing prevalence and its profound impact on the major diseases of the western world require that clinicians consider its diagnosis and management on a routine basis. Recently published guidelines on its definition now make convenient and reliable diagnosis possible. Also, there is new and better understanding of the complex dyslipidemias and other risk factors strongly associated with the metabolic syndrome, which greatly increase the risk of clinical atherosclerotic events. Comprehensive clinical evaluation of these dyslipidemias and associated atherosclerosis risk factors can lead to their aggressive treatment, customized according to the circumstances of each patient. These steps are now more feasible and more clearly desirable than ever before. Statins alone greatly reduce atherosclerosis risk, but combination lipid therapy is often required for optimal dyslipidemia management and atheroprevention. PMID- 12643149 TI - Epidemiology of the insulin resistance syndrome. AB - The insulin resistance syndrome consists of the co-occurrence of metabolic risk factors for type 2 diabetes and cardiovascular disease, including overall obesity, central obesity, dyslipidemia (characterized by elevated levels of triglycerides and low levels of high-density lipoprotein cholesterol), hyperglycemia, and hypertension. Using criteria proposed by the National Cholesterol Education Program Adult Treatment Panel III, national survey data suggest the insulin resistance syndrome is very common, affecting about 24% of US adults aged greater than 20 years. The syndrome is more common in older people and in Mexican Americans, and will increase in prevalence as populations age and become more obese. Identification of the syndrome warrants aggressive interventions known to prevent type 2 diabetes and cardiovascular disease, including weight reduction, increased physical activity, and control of hypertension and dyslipidemia. PMID- 12643152 TI - Diabetes and depression. PMID- 12643151 TI - Endothelial dysfunction and the metabolic syndrome. AB - The metabolic syndrome is a highly prevalent multifaceted clinical entity produced through the interaction of genetic, hormonal, and lifestyle factors. A distinctive constellation of abnormalities precedes and predicts the accelerated development of inflammation and coagulation represent emerging risk contributors associated with obesity and insulin resistance, central components of the metabolic syndrome, which act in concert with traditional abnormalities to increase cardiovascular risk. The initiation and progression of atherosclerosis may have its origins in impaired endothelial function that can be detected at the earliest stages of development of the syndrome. The basic elements of the metabolic syndrome and accelerated phase of atherogenesis are often silent partners that present many years before the onset of type 2 diabetes mellitus. The ability to detect and monitor subclinical vascular disease, as a reflection of the multiple factors that contribute to impair arterial wall integrity, holds potential to further refine cardiovascular risk stratification. Noninvasive assessment of vascular health may also aid the clinical decision-making process by guiding therapeutic interventions to optimize vascular protection in the metabolic syndrome. PMID- 12643150 TI - Association of albuminuria and the metabolic syndrome. AB - Microalbuminuria clusters with the metabolic syndrome, and both conditions predict cardiovascular disease mortality. The reported relationships of microalbuminuria with the individual components of the metabolic syndrome (i.e., hyperglycemia, insulin resistance, hypertension, dyslipidemia, abdominal obesity) are variable. Each of these components, as well as intrauterine effects and diet and other lifestyle factors, may contribute to elevated risk of microalbuminuria in certain population groups. Recent evidence indicates a role for oxidation and inflammation in cardiovascular disease, and endothelial dysfunction (exacerbated by factors such as dyslipidemia) may be the mediator of this relationship. Because endothelial dysfunction can also be manifested as microalbuminuria, this provides a potential explanation of the observed association of the metabolic syndrome, chronic inflammation, and microalbuminuria. PMID- 12643153 TI - The epidemiology of neuropathic foot ulcers in individuals with diabetes. AB - There is now substantial evidence that diabetic neuropathy is a major etiologic factor for diabetic foot ulcers. Epidemiologic data show a very strong association between foot ulcers and indicators of sensorimotor neuropathy. Additionally, a causal pathway (sensorimotor neuropathy, which leads to increased pressures at foot surfaces, which then leads to foot ulcers) has been postulated. Evidence suggests that other factors also contribute to foot ulcer risk through an interaction with neuropathy such as foot deformity, footwear, and body weight. There has been an effort to optimize modalities for screening diabetic patients for hypoesthesia in order to prevent foot ulcers; however, there have not been definitive prospective studies that have compared the effectiveness of available tests. PMID- 12643155 TI - Focal and multifocal neuropathies. AB - The focal and multifocal neuropathies affect only a minority of patients with diabetes; however, they form a major clinical problem in terms of diagnosis, development of significant symptoms and signs, and often inadequate therapy. Diagnosis requires accurate and detailed clinical history and neurologic examination combined with targeted neurophysiologic tests, which differ considerably from those carried out in day-to-day practice. Because of their relatively infrequent occurrence, treatment is not evidence based. PMID- 12643154 TI - Future treatments for diabetic neuropathy: clues from experimental neuropathy. AB - Neuropathy remains a major complication of diabetes and there is no approved treatment that prevents its progression or alleviates the associated symptoms. Animal models of diabetic neuropathy are hampered by their short life span, which precludes the development of overt structural pathology, and they are best viewed as exhibiting early metabolic, neurochemical, and functional indices of nerve disorders that may predict progression to overt diabetic neuropathy. In this context, diabetic animals have use in both establishing potential etiologic mechanisms and for screening novel therapeutic agents. Treatment strategies are evolving in concert with a developing understanding of how hyperglycemia causes nerve dysfunction and recent or ongoing clinical trials are investigating this rational approach to drug design. It is only by the successful demonstration of clinical efficacy of a compound developed by this approach that the use of animal models of diabetic neuropathy can be validated. PMID- 12643156 TI - Diagnosis and management of diabetic neuropathy. AB - Diabetic sensorimotor polyneuropathy (DSP) remains the most common microvascular complication of both type I and type 2 diabetes, and poses a unique set of management challenges in the prevention of foot complications. Although different quantitative tests are available, the preliminary diagnosis of DSP can be reliably made using simple and rapid screening tests in the family physician's office or in the diabetes clinic. The Semmes-Weinstein 10-g monofilament examination is a popular, simple clinical modality for the prediction of early DSP, foot ulceration, and amputation, and, in turn, a predictor of mortality in patients with diabetes. The management of DSP is centered on optimal glycemic control, diligent foot care, and pain control as a means of preventing the progression of DSP and reducing the morbidity associated with foot complications. PMID- 12643157 TI - Etiology and treatment of erectile failure in diabetes mellitus. AB - Men with diabetes are especially prone to neuronal and endothelial disease and are afflicted with erectile dysfunction (ED) at a much higher incidence and prevalence than normal men, with a consequent reduction in quality of life. ED in diabetes results from combinations of impairments from nearly every step in the production of a penile erection. These include the failed transmission of neural signals to and from the spinal cord due to neuropathy resulting in reduced neural nitric oxide (NO) delivery to cavernosal smooth muscle, impaired sinusoidal endothelial cell NO release because of endothelial dysfunction, reduced arterial and arteriolar inflow due to peripheral vascular disease, and failure of relaxation of the corpora from glycation of the elastic fibers. The evaluation of men with diabetes and ED is similar to that in nondiabetic men. It includes assessment of gonadal, neural, arterial, and on occasion, venous function, along with marital and psychological status. Although treatment options for diabetic men with ED have fortunately expanded, the risk factors for ED are the same as those for cardiovascular disease; a flagging penis should raise a red flag of warning to evaluate and treat the risk factors for coronary, cerebral, and peripheral vascular disease, which are common in persons, of either sex, afflicted by diabetes. PMID- 12643158 TI - Healing the diabetic wound and keeping it healed: modalities for the early 21st century. AB - The incidence of lower extremity wounds, infections, and amputations continues to increase worldwide. Fortunately, the past decade has seen a surge of activity in research into new modalities in wound healing and, even more recently, prevention. This article discusses the basic physiology of wound healing, with particular attention being paid to current and potential wound healing modalities. These include, among other modalities, discussions of bioengineered skin, hyaluronic acid, subatmospheric pressure therapy, growth factors, and endogenous cytokine stimulation. Emerging concepts targeting prevention through novel methods of patient empowerment, dermal thermometry, and computerized activity monitoring are also discussed. PMID- 12643159 TI - The relationship between glucose control and the development and progression of diabetic nephropathy. AB - Diabetic nephropathy is a major cause of morbidity and mortality in patients with diabetes; it occurs in about one third of such patients. The course of nephropathy is better defined and similar for both type 1 and type 2 diabetes. Patients initially develop microalbuminuria (albumin excretion rates [AERs] between 20 and 200 micrograms/min), then overt nephropathy (AER > or = 200 micrograms/min), and finally a decline in glomerular filtration rate (GFR) eventuating in end-stage renal disease. Although metabolic control has long been hypothesized as a contributor to the development of nephropathy, it is only in recent years that this hypothesis has been proven. A number of observational studies have shown correlations between glycemic control and the development of various levels of albuminuria and also declines in GFR. However, large long-term prospective, randomized, interventional studies have now definitely proven that improved metabolic control that achieves near-normoglycemia can significantly decrease the development and progression of diabetic nephropathy as well as other long-term complications of diabetes, including retinopathy and neuropathy. It is now conceivable that the achievement of near-normoglycemia, plus medications that inhibit the renin-angiotensin system if microalbuminuria develops, may greatly decrease the numbers of patients eventually requiring renal replacement therapy. PMID- 12643160 TI - Blood pressure control--effects on diabetic nephropathy progression: how low does blood pressure have to be? AB - Hypertension and diabetes are independent risk factors for both cardiovascular disease and renal complications. Coexistence of these comorbid conditions predisposes the patient to a much greater risk of progression to end-stage renal disease. Combined with the increased cardiovascular mortality, this has led to recent Joint National Committee-VI recommendations for the initiation of antihypertensive therapy for people with diabetes at a blood pressure of 130/85 mm Hg, a level lower than that recommended for the nondiabetic population. Results of a review of recently published investigations on the effects of blood pressure on diabetic nephropathy progression are presented in this article. This review finds evidence to support reducing the mean arterial blood pressure to levels below 95 mm Hg, a level that is even lower than the blood pressure of 130/80 mm Hg (mean arterial pressure of 97 mm Hg) recommended by the American Diabetes Association and National Kidney Foundation. The effect of blood pressure on renal disease progression is linear and appears to have no lower threshold for the benefits of blood pressure reduction on limiting nephropathy progression. The answer to the question of how low does blood pressure have to be to minimize the effects of blood pressure on diabetic nephropathy progression might be "the lower, the better." PMID- 12643161 TI - Circadian changes in blood pressure and their relationships to the development of microalbuminuria in type 1 diabetic patients. AB - Diabetic nephropathy in type I diabetic patients, as it is currently understood, progresses in a stepwise fashion from normoalbuminuria to microalbuminuria, then to overt proteinuria and progression to chronic renal failure, and ultimately to end-stage renal disease. The role of early blood pressure changes in relation to diabetic nephropathy is now better understood in light of recent data using ambulatory blood pressure monitoring as a means to monitor blood pressure changes noninvasively throughout the day. Cross-sectional studies with type I diabetic patients with microalbuminuria have shown that the normal nocturnal blood pressure often fails to fall normally during sleep. The question of which comes first, microalbuminuria or a rise in blood pressure in patients with type I diabetes, was recently addressed in a prospective study. An increase in systolic blood pressure during sleep precedes the development of microalbuminuria and may play a causative role in its development. PMID- 12643163 TI - Pulmonary function in diabetes mellitus. PMID- 12643162 TI - Appropriate drug therapy for improving outcomes in diabetic nephropathy. AB - Diabetic nephropathy is the leading cause of end-stage kidney disease in the United States. The majority of these cases are attributed to those with type 2 diabetes. Elevated blood pressure, proteinuria, and increased activity of the renin-angiotensin-aldosterone system (RAAS) play a major role in the development and progression of chronic kidney disease attributed to diabetes mellitus. Moreover, drugs that inhibit angiotensin II synthesis or block the angiotensin II type I receptor lower blood pressure, reduce proteinuria, and improve outcomes in patients with chronic kidney disease caused by diabetes. This article highlights improvements in the current management of diabetic nephropathy afforded by agents that inhibit the RAAS, discusses their limitations, and considers novel strategies to prevent onset and progression of diabetic nephropathy. Current opinions concerning combination drug therapy with agents that block the RAAS at multiple sites, as well as combining calcium channel blockers with either angiotensin-converting enzyme inhibitors or angiotensin II receptor antagonists, are also discussed. PMID- 12643164 TI - Role of inflammatory pathways in the development and cardiovascular complications of type 2 diabetes. AB - Experimental and epidemiologic studies support the role of inflammation in the development of type 2 diabetes and atherosclerosis. Serum levels of inflammatory markers, in particular highly sensitive C-reactive protein, have been found to be strong predictors of increased risk for type 2 diabetes and cardiovascular disease independent of traditional risk factors. A beneficial effect of thiazolidinediones, angiotensin-converting enzyme inhibitors, and statins in the prevention of type 2 diabetes and cardiovascular events has recently been reported, and potential anti-inflammatory mechanisms of action for these compounds have been described. Prospective, randomized clinical trials are currently underway to confirm these initial findings and define indications for treatment of patients at risk. PMID- 12643165 TI - Insulin analogues: new therapies for type 2 diabetes mellitus. AB - Insulin therapy is ultimately necessary for the control of blood glucose in a majority of patients with type 2 diabetes mellitus. Unfortunately, the pharmacokinetic characteristics of previously available rapid-, intermediate-, and long-acting preparations make sustained normoglycemia almost impossible. Advances in molecular genetic engineering have made possible the development of insulin analogues with pharmacokinetics that more closely mimic the needs of patients with type 2 diabetes. In the following article, we explore the insulin analogues currently available for clinical use, their pharmacokinetics, and the rationale for their use in the treatment of type 2 diabetes, and follow-up with a brief examination of future developments. PMID- 12643166 TI - Antiobesity drugs: current and future issues. AB - Overweight and obesity affect more than 60% of the population in the United States. Many social and environmental factors influence this phenomenon. Because of the tremendous difficulties associated with addressing these factors, health care providers have been looking for a magic pill that resolves this tissue. Although there is no panacea, this article is intended to concisely review commonly prescribed medications for obesity, their efficacy, and side effects. In addition, some pharmacologic agents with a promising antiobesity action are discussed. PMID- 12643167 TI - Preventing diabetes by treating aspects of the metabolic syndrome. AB - The metabolic syndrome often develops into and is usually present in type 2 diabetes in association with premature cardiovascular disease. Treating diabetes can prevent some of its devastating consequences, but it does not eliminate them all. With the goal to eliminate all the adverse consequences of the syndrome, the optimal approach would be through its prevention. Insulin resistance appears to be pivotal to development of the syndrome complex that includes features such as intra-abdominal or visceral obesity, hypertension, impaired glucose homeostasis, dyslipidemia with elevated triglycerides and low high-density lipoprotein without elevations of low-density lipoprotein, a procoagulant state, and impaired vascular function. Improving the insulin resistance needs to be the primary target of the therapy. Hyperglycemia, which is one feature of the metabolic syndrome, may range from impaired glucose tolerance (IGT) to overt diabetes. The risk of progression of the disease from IGT to diabetes is increased with time and the presence of various risk factors. Diabetes is a disease of serious concern because of the associated complication of the disease and the huge impact on the health care costs. Many short- and longer-term trials have shown promise in the prevention of diabetes and its metabolic and cardiovascular consequences. PMID- 12643168 TI - The choice of antihypertensive drugs in patients with diabetes: angiotensin II and beyond. AB - Affecting over 16 million individuals, diabetes mellitus is among the leading causes of mortality in the United States. Hypertension is a common finding among diabetic patients and increases their morbidity and mortality. Control of blood pressure in this population has been shown to improve outcomes. Recent randomized trials have proven the benefit of lower blood pressure goals in the treatment of hypertensive patients as compared with nondiabetic patients. Randomized controlled trials have also demonstrated that the selection of the antihypertensive agent used to treat hypertension in diabetic patients is as important as the reduction of blood pressure levels to the recommended levels. In this article, we first focus on the importance of the renin angiotensin system in the development of diabetic complications, and then we review the results of the recent studies that have had a major impact on the treatment of hypertension in diabetes. PMID- 12643169 TI - The influence of dietary fat on insulin resistance. AB - Dietary fat has been implicated in the development of insulin resistance in both animals and humans. Most, although not all, studies suggest that higher levels of total fat in the diet result in greater whole-body insulin resistance. Although, in practice, obesity may complicate the relationship between fat intake and insulin resistance, clinical trials demonstrate that high levels of dietary fat can impair insulin sensitivity independent of body weight changes. In addition, it appears that different types of fat have different effects on insulin action. Saturated and certain monounsaturated fats have been implicated in causing insulin resistance, whereas polyunsaturated and omega-3 fatty acids largely do not appear to have adverse effects on insulin action. Given the importance of insulin resistance in the development of diabetes and heart disease, establishing appropriate levels of fat in the diet is an important clinical goal. PMID- 12643170 TI - Cardiovascular fitness and type 2 diabetes. AB - Exercise has long been recognized as a cornerstone in diabetes care. Even so, modern science is just beginning to understand the physiologic impact cardiovascular fitness has on long-term diabetes management, and the prevention or delay, of type 2 diabetes. Recent advances in research are defining the metabolic pathways involved in insulin signaling and in reducing insulin resistance. Tools and guidelines exist for the health care provider to use when prescribing cardiovascular activities to individuals with type 2 diabetes. The true challenge now lies in assisting individuals in implementing and maintaining safe, therapeutic levels of physical activity. PMID- 12643171 TI - Antioxidant vitamins and their influence in diabetes mellitus. AB - Diabetes mellitus is a chronic disease associated with serious complications. A number of studies have suggested that enhanced oxidation is the underlying abnormality responsible for some of the complications of diabetes. It is not known whether the ingestion of antioxidant vitamins could retard or perhaps reverse the oxidative damage. The information regarding the benefit of antioxidant vitamin supplementation is conflicting some trials have demonstrated adverse effects of excessive consumption of vitamin supplements. In this article, we review the available literature on the association of cardiovascular events and ingestion of vitamins with antioxidant properties. Given the lack of data to substantiate the benefit and safety of ingestion of antioxidant vitamins in excess of the recommended dietary allowance, physicians should avoid the recommendation of vitamin supplementation to their patients. PMID- 12643173 TI - The intriguing diversity of the glucagon gene products. PMID- 12643172 TI - Protein and diabetes: much advice, little research. AB - In persons with diabetes, moderate hyperglycemia can contribute to an increased turnover of protein. To maintain body composition and nitrogen balance requires metabolic control and sufficient protein and energy intakes. However, because most adults eat at least 50% more protein than is required, people with diabetes appear to be protected from protein malnutrition when consuming a usual diet. Although nonessential amino acids undergo hepatic gluconeogenesis, peripheral glucose concentrations do not increase after protein ingestion. The fate of produced glucose is unknown. Protein does not contribute to sustained elevations of glucose levels, slow absorption of carbohydrate, or help in the treatment of hypoglycemia--advice often given to persons with diabetes. Protein is, however, just as potent a stimulant of insulin secretion as glucose. No long-term research is available to document that high-protein, low-carbohydrate diets are continued long-term or that weight lost initially is maintained better from these diets than from traditional weight loss diets. Furthermore, in persons with type 2 diabetes, weight loss is reported to be related to energy restriction and not to the protein-to-carbohydrate ratio of the diet. PMID- 12643174 TI - Action for health in diabetes: the look AHEAD clinical trial. PMID- 12643175 TI - Hepatic steatosis and type 2 diabetes mellitus. AB - Type 2 diabetes is strongly associated with nonalcoholic fatty liver disease (NAFLD), a spectrum of liver damage that ranges from relatively benign hepatic steatosis to potentially fatal cirrhosis. The severities of insulin resistance and liver damage parallel each other, with the greatest prevalence of cirrhosis occurring in cirrhotics. However, it is unknown whether one of these conditions causes the other, or if both are consequences of another process. Experimental evidence suggests that both insulin resistance and NAFLD result from a chronic inflammatory state. The mechanisms driving this chronic inflammation are unknown but might include the egress of products from intestinal bacteria into the portal blood, liver, and systemic circulation to trigger a sustained inflammatory cytokine response in genetically susceptible individuals. More research is needed to evaluate this hypothesis and to determine the benefits of treatments that interrupt this pathogenic cascade. PMID- 12643176 TI - Skeletal muscle triglyceride: marker or mediator of obesity-induced insulin resistance in type 2 diabetes mellitus? AB - The inability of insulin to stimulate glucose metabolism in skeletal muscle is a classic characteristic of type 2 diabetes, but this insulin resistance entails altered patterns of lipid metabolism as well. An association between intracellular triglyceride and insulin resistance has been well established in both human and animal studies of obesity-related insulin resistance and type 2 diabetes. Skeletal muscle's ability to select substrate for fuel metabolism, a metabolic flexibility, is also lost in insulin resistance, and defects in fatty acid metabolism during fasting or postabsorptive conditions likely play an important role in lipid oversupply to insulin-resistant muscle. These impairments appear to be at least indirectly centered on the ability of mitochondria to oxidize fatty acids, possibly through mediation of lipid metabolite levels such as ceramide or diacylglycerol, which are known to directly attenuate insulin signaling. Moreover, periodic use of muscle triglyceride by exercise may mediate the association between muscle triglyceride and insulin resistance. Thus, it appears that skeletal muscle triglyceride is perhaps a surrogate for other lipid species having a more direct effect on insulin action. Defining mechanisms by which dysregulation of fatty acid metabolism and persistent lipid oversupply alter insulin action may help to target more effective strategies to prevent or treat type 2 diabetes. PMID- 12643178 TI - Is hepatic glucose production increased in type 2 diabetes mellitus? AB - Based on recent studies, including our own, using what we consider to be an appropriate technique to estimate rates of hepatic glucose production (HGP), this article can be summarized as follows: 1) HGP in the overnight fasted state is near normal in obese type 2 diabetes (T2D) subjects, i.e., it may be increased by a mean of 12% compared to matched control subjects. 2) Suppression of HGP by insulin shows a rightward shift of the dose response curve (reduced insulin sensitivity) but normal maximal suppression (no maximum velocity defect). 3) In the overnight fasted state, gluconeogenesis is responsible for two thirds of HGP in T2D subjects and is about 5% to 10% increased compared to healthy subjects. 4) Suppression of HGP during a meal is close to normal. 5) The slightly increased HGP values throughout the 24-hour period together with reduced metabolic clearance rate (peripheral insulin resistance) and increased carbohydrate intake is responsible for the increase in fasting plasma glucose values. 6) Hypothetically, the role of the liver in nondiabetic and T2D subjects may be to produce the amount of glucose needed for metabolism in peripheral tissues. If insulin-mediated glucose uptake in skeletal muscle is reduced, plasma glucose will increase due to the "nonsuppressed" HGP values. Plasma glucose continues to rise until glucose-mediated glucose uptake compensates completely for the reduction in insulin-mediated glucose uptake. PMID- 12643177 TI - Emerging paradigms for understanding fatness and diabetes risk. AB - It is widely accepted that increasing adiposity is associated with insulin resistance and increased risk of type 2 diabetes. The predominant paradigm used to explain this link is the portal/visceral hypothesis. This hypothesis proposes that increased adiposity, particularly in the visceral depots, leads to increased free fatty acid flux and inhibition of insulin action via Randle's effect in insulin-sensitive tissues. Recent data do not entirely support this hypothesis. As such, two new paradigms have emerged that may explain the established links between adiposity and disease. PMID- 12643179 TI - Role of the kidney in hyperglycemia in type 2 diabetes. AB - Overproduction of glucose is the major factor responsible for fasting hyperglycemia in type 2 diabetes. Formerly, this had been considered to be solely due to excessive hepatic glucose production because the human kidney was not regarded as an important source of glucose except during acidosis and after prolonged fasting. However, data accumulated over the last 60 years in animal and in vitro studies have provided considerable evidence that the kidney plays an important role in glucose homeostasis in conditions other than acidosis and prolonged fasting. This article summarizes early work in animals and humans, discusses methodologic issues in assessing renal glucose release in vivo, and provides evidence from recent human studies that the kidney substantially contributes to glucose overproduction in type 2 diabetes. PMID- 12643180 TI - Progression from IGT to type 2 diabetes mellitus: the central role of impaired early insulin secretion. AB - Impaired glucose tolerance (IGT) is characterized by plasma glucose responses to an oral glucose challenge that are above normal but not at the level defining diabetes. IGT is a common condition that greatly increases risk for the subsequent development of type 2 diabetes. Individuals with IGT manifest abnormalities in both insulin action and early insulin secretion similar to those seen in patients with type 2 diabetes. These abnormalities not only precede diabetes, they predict it as well. Furthermore, the progression from IGT to diabetes is characterized by a dramatic decline in early insulin secretion. It is now evident that early insulin secretion plays an important role in the rapid and efficient suppression of endogenous glucose production following a meal. Loss of early insulin secretion initially leads to postprandial hyperglycemia which, as the disease progresses, worsens to clinical hyperglycemia. Obesity and a high fat diet may contribute to the development of both insulin resistance and insulin secretory dysfunction in susceptible individuals. Strategies that improve insulin resistance and enhance early insulin secretion may prevent the progression from IGT to diabetes. Already, there is substantial evidence the weight loss and exercise may reduce the risk of developing diabetes by up to 58%. Other trials using pharmacologic agents to decrease insulin resistance and increase early insulin secretion are underway. Prevention remains the best hope for a long-term solutions to the worldwide epidemic of diabetes. PMID- 12643181 TI - Role of ACE inhibitors in treating hypertensive diabetic patients. AB - Cardiovascular disease (CVD) is a major determining factor of morbidity and mortality in type 2 diabetic patients. Hypertension, which accompanies diabetes in more than 70% of cases, contributes to increased prevalence of CVD events in this group of patients. Results from the United Kingdom Prospective Diabetes Study (UKPDS) indicated that reduction of elevated blood pressure might decrease CVD morbidity and mortality more than reduction of hyperglycemia. Activation of circulating and tissue renin-angiotensin system (RAS) contributes to the development of both hypertension and insulin resistance in patients with the cardiometabolic syndrome. Angiotensin-converting enzyme (ACE) inhibitor therapy in patients with the cardiometabolic syndrome may improve insulin action as well as lessen CVD. In clinical trials, ACE inhibitors have been shown to be more efficient than other antihypertensive medications (i.e., calcium channel blockers) in the reduction of CVD morbidity and mortality in hypertensive diabetics. In this article, we summarize possible mechanisms by which ACE inhibition may improve insulin resistance, coagulation/clotting, and vascular function abnormalities, and postpone or even prevent the development of type 2 diabetes in hypertensive patients. PMID- 12643182 TI - Microalbuminuria in diabetes: focus on cardiovascular and renal risk reduction. AB - Albuminuria has long been recognized as a harbinger for the progression of diabetic nephropathy. Recently, it has become increasingly recognized that albuminuria is a powerful risk factor for cardiovascular disease and stroke. In this article we explore evidence for albuminuria being an integral component of the cardiometabolic syndrome and a risk factor for cardiovascular disease and stroke. PMID- 12643184 TI - Heart protection study: LDL lowering therapy and cardiovascular outcomes in patients with low cholesterol levels or diabetes. PMID- 12643183 TI - Cardiovascular risk factors in diabetic patients with hypertension. AB - Individuals with diabetes mellitus have cardiovascular disease (CVD) mortality comparable to nondiabetics who have suffered a myocardial infarction or stroke. Aggressive management of risk factors such as hypertension, dyslipidemia, and platelet dysfunction in persons with diabetes has been shown to reduce morbidity and mortality in prospective randomized controlled clinical trials. Accordingly, there are national mandates to lower blood pressure to less than 130/85 mm Hg, reduce low-density lipoprotein cholesterol to less than 100 mg/dL, and institute aspirin therapy in adult patients with diabetes. Although not definitively shown to reduce CVD, there are also recommendations to control the level of glycemia, as well. This article discusses CVD risk factors in the diabetic patient with hypertension. PMID- 12643185 TI - The epidemiology of atherosclerosis and its risk factors among Native Americans. AB - In the past, the rates of risk factors for atherosclerosis and cardiovascular disease (CVD) as well as the manifestations of coronary heart disease, stroke, and peripheral vascular disease in Native Americans have been relatively low compared to the general United States population. However, over the past several decades the rates of these CVD-associated risk factors have markedly increased with the concomitant development of a significant and alarming rise in the manifestations of atherosclerosis. PMID- 12643188 TI - Hemoglobin A1c is not the only issue. PMID- 12643186 TI - The role of non-LDL:non-HDL particles in atherosclerosis. AB - Elevated concentrations of circulating apolipoprotein B (apoB)-containing lipoproteins, other than low-density lipoprotein (LDL), have been implicated as causative agents for the development of atherosclerosis. A form of dyslipidemia, the atherogenic lipoprotein profile, that consists of elevated intermediate density lipoprotein (IDL), triglycerides (TGs), dense LDL and dense very low density lipoprotein (VLDL), and low high density lipoprotein-2, occurs in 40% to 50% of patients with coronary artery disease (CAD). The recently released Adult Treatment Panel III guidelines suggest that because elevated TGs are an independent CAD risk factor, some TG-rich lipoproteins, commonly called remnant lipoproteins, must be atherogenic. Relevant to this series on diabetes, a number of studies have shown that in type 2 diabetes, the severity of CAD is positively related to the numbers of TG-rich particles in the plasma. Although less clear, other studies in type 2 diabetes suggest that elevated levels of lipoprotein (a) [Lp(a)] may also be independently associated with CAD. In this article, we summarize evidence for the role of apoB-containing lipoprotein particles other than LDL in the development of atherosclerosis and discuss methods of quantification and possible pharmacologic interventions for lowering their plasma concentrations. The particles reviewed include the TG-rich lipoproteins: VLDL and its remnants, chylomicron remnants and IDL, and the C-rich lipoprotein: Lp(a). PMID- 12643187 TI - The effect of dietary intervention on serum lipid levels in type 2 diabetes mellitus. AB - Dietary therapy is the cornerstone of lipid management in patients with type 2 diabetes mellitus. The key strategies are the reduction of intake of saturated fat, trans unsaturated fat and cholesterol, and the reduction of energy intake to promote weight loss. This approach will produce significant improvements in the serum levels of low-density lipoprotein (LDL) cholesterol, triglycerides, and high-density lipoprotein (HDL) cholesterol. According to both the American Diabetes Association and the National Cholesterol Education Program (NCEP), the primary target of therapy is the serum LDL cholesterol level, with the secondary targets being non-HDL cholesterol, triglycerides, and HDL cholesterol. The recently updated guidelines of the NCEP place new emphasis on increasing soluble fiber intake to 10 to 25 g/d and adding foods fortified with plant stanols/sterols (2 g/d) as options to enhance the LDL cholesterol-lowering effect of diet. PMID- 12643189 TI - Vascular reactivity in diabetes mellitus. AB - Cardiovascular disease is responsible for 70% of the deaths in diabetes mellitus. Endothelial dysfunction occurs early in atherosclerosis and can now be assessed accurately and reproducibly by noninvasive means. Hyperglycemia, insulin resistance, microvascular complications, and coexistent conditions have been shown to impair endothelial function in diabetes. Resistance to insulin's vasodilatory effect and increased free radical generation are probable mechanisms responsible for the endothelial dysfunction in this state. Improvement in glycemic control, insulin and insulin sensitizers, cholesterol lowering, hypolipidemic agents, angiotensin-converting enzyme inhibitors, and angiotensin II receptor blockers have improved clinical cardiovascular outcomes and have also been shown to improve endothelial function. Strategies designed to assess and improve endothelial function may be further beneficial in terms of cardiovascular outcomes in diabetes mellitus, and need to be tested in a clinical setting. PMID- 12643190 TI - Endothelium, inflammation, and diabetes. AB - The endothelium has several diverse functions in maintaining vascular integrity in terms of structure and function. Two key vasodilators, nitric oxide (NO) and prostacyclin, maintain the vascular pathway, inhibit platelet aggregation, and are antithrombotic. More recently, they have been shown to be anti-inflammatory, and thus are potentially antiatherogenic. It has recently been noted that insulin stimulates NO release by the endothelium. Insulin is a vasodilator, has antiplatelet activity, and is anti-inflammatory. Similar anti-inflammatory effects of thiazolidinediones (TZDs), troglitazone and rosiglitazone, suggest that they too may have potential antiatherogenic effects. These effects of insulin and TZDs are important because the two major states of insulin resistance, obesity and type 2 diabetes, are associated with a marked increase in atherosclerosis coronary heart disease, and stroke. These recent observations have extremely momentous implications for the understanding of the pathogenesis of atherosclerosis in insulin-resistant states and for a rational approach to their comprehensive treatment, including the prevention of atherosclerosis and its complications. PMID- 12643191 TI - Mechanisms involved in platelet hyperactivation and platelet-endothelium interrelationships in diabetes mellitus. AB - This article discusses the recent findings about the mechanisms involved in platelet hyperactivation in diabetes mellitus. Particular attention is focused on the advances pertaining to 1) new features of platelet activation; 2) the role of calcium fluxes; 3) the role of glucose, insulin, and insulin resistance; 4) the role of nitric oxide; and 5) the role of endothelium. PMID- 12643192 TI - Does pregnancy accelerate the rate of progression of diabetic retinopathy? AB - Pregnancy may be associated with a number of ocular changes, including the development of new ocular conditions or modifications of existing conditions. The most common ocular condition modified by pregnancy is diabetic retinopathy. Pregnancy is associated with an increased risk of development and progression of diabetic retinopathy. The factors associated with its progression include the pregnant state itself, duration of diabetes, amount of retinopathy at conception, blood glucose control, and the presence of coexisting vascular disease. Although the rate of regression of diabetic retinopathy at the end of pregnancy or the postpartum period is high, careful monitoring of these patients is necessary to optimize both the vision and pregnancy outcomes. PMID- 12643193 TI - The utility and efficacy of the new insulins in the management of diabetes and pregnancy. AB - The quest for tight glucose control has led to the development of insulin analogues (insulin lispro, insulin aspart, insulin glargine) designed to optimize glucose control while minimizing the impact of insulin therapy on daily life. Women with diabetes are increasingly likely to become pregnant on these new products and to benefit from their use while pregnant. Caution over the use of new medications in pregnancy has limited experience, but increasing evidence is emerging that use of insulin lispro is not associated with excess risk of fetal malformations, is associated with similar or better outcomes during pregnancy, and is preferred over regular insulin. Similar evidence for the other insulin analogues has yet to emerge, but is unlikely to be dissimilar. PMID- 12643194 TI - Current controversies in the mechanisms and treatment of gestational diabetes. AB - Gestational diabetes mellitus (GDM) is a heterogeneous entity, including carbohydrate intolerance of variable severity with onset or first recognition during pregnancy. Insulin resistance and beta-cell dysfunction are thought to be major determinants of its development. Its pathomechanism in many ways resembles that of type 2 diabetes. There is an evolving body of evidence from the last decade presenting similarities between gestational diabetes and the metabolic (insulin resistance) syndrome. These new observations suggest that GDM might be an early manifestation of the metabolic syndrome. The desired treatment target of GDM is normoglycemia. It can be reached by dietary treatment; however, if it fails, maternal glycemic monitoring or combined fetal-maternal monitoring, or even insulin (if required) can help reach it. Multiple daily insulin regimens are becoming more widely accepted for the treatment of GDM. Insulin analogues, however, need some more evidence to support their usefulness and safety during pregnancy. The screening for GDM, the reclassification, regular care, and follow up of these women after pregnancy are of the utmost importance to delay or prevent not only type 2 diabetes but cardiovascular complications as well. PMID- 12643195 TI - Autoimmunity and familial risk of type 1 diabetes. AB - There is evidence that the process leading to type I diabetes may start in early infancy or already in utero. Even though diabetes-associated antibodies can be detected in up to half of the pregnancies of mothers with type I diabetes, pregnancy itself has no major effect on these antibodies. If such antibodies are present in the mother, they are transferred to the fetal circulation and are detectable in cord blood. Most of the transplacentally transferred antibodies disappear by 6 months of age, but may persist even longer. Antibodies present in cord blood may represent true induction of beta-cell autoimmunity, but such a phenomenon is extremely rare. The offspring of affected mothers have a 2% to 3% risk of type I diabetes, which is about one third of that in the offspring of affected fathers. A novel conceivable explanation is that exogenous insulin transplacentally transferred in immune complexes might lead to the induction of tolerance to insulin, which may be the primary autoantigen in type I diabetes. The possible protective or predisposing effect of diabetes-associated antibodies detectable at birth on progression to clinical type I diabetes later will be assessed in ongoing prospective birth cohort studies. PMID- 12643196 TI - Pancreas and kidney transplantation. AB - Kidney transplantation is preferred over dialysis for management of end-stage renal disease complicating type I or type 2 diabetes, for those who are eligible. Simultaneous pancreas-kidney (SPK) or pancreas after kidney transplantation (PAK) is an important alternative to kidney transplantation alone for type I diabetes patients if the patient is able to withstand the additional risks of these procedures, because of the benefits of glucose control on other diabetic complications. Pancreas transplantation alone (PTA) is most useful for the treatment of debilitating, frequent hypoglycemia complicating type I diabetes, if renal function is adequate. One-year pancreas graft survival is best after SPK (82%) but has significantly improved after both PAK (74%) and PTA (76%). The I year kidney graft and patient survival rates after SPK are similar to kidney transplantation alone. Pancreas transplantation normalizes glucose beyond what can be achieved with insulin therapy and has been shown to decrease progression of or improve most, if not all, diabetic end-organ complications using current immunosuppression regimens. However, the diabetologist and endocrinologist should remain involved in the care of the pancreas or kidney transplant recipient for treatment of vascular disease risk factors such as dyslipidemia, surveillance of other diabetic complications including foot ulcers, surveillance and treatment of bone loss, and management of hyperglycemia if it recurs. PMID- 12643197 TI - Islet transplantation: travels up the learning curve. AB - Great excitement was generated in 2000 by a report from the University of Alberta in Edmonton, Canada, that seven of seven type I diabetic patients transplanted with intrahepatic cadaveric islets were normal glycemic, 1-year post transplantation without the use of exogenous insulin treatment. The follow-up information from the same researchers with a larger group of patients indicated that in a group of 12 alloislet recipients, five had impaired glucose tolerance and three had post-transplantation diabetes. Great attention is now being directed toward understanding why alloislet recipients who are initially successful may later develop partial failure. At the same time, the Immune Tolerance Network is sponsoring a multicenter trial using the Edmonton protocol to ascertain whether these results can be replicated by other transplant groups in the United States, Canada, and Europe. Detailed studies of islet beta-cell function have revealed intact insulin secretion in autoislet and alloislet transplant recipients. In contrast, glucagon responses to insulin-induced hypoglycemia are absent from islets transplanted intrahepatically; however, alpha cells within intrahepatic islets are capable of releasing glucagon in response to intravenous arginine. Although many technical refinements are underway to make this procedure even more efficacious, supply and demand issues are a major concern and must be dealt with before the procedure of islet transplantation can be considered generally available for patients with diabetes. PMID- 12643198 TI - Alternatives to unmodified human islets for transplantation. AB - Islet transplantation as a procedure to induce insulin independence is still a long way from benefitting the population of more than I million type I diabetic patients in the United States. In addition to the problems involved with immune suppression, the most significant obstacle is a scarcity of human organs for transplantation. In 1999, only 5882 donated pancreases were available, of which only 50% could be expected to produce islet yields suitable for clinical purposes. In this article, we review various sources with the potential to provide tissue for transplantation. These sources include islet and nonislet cells derived from both human and nonhuman sources, with an emphasis on human cells. PMID- 12643199 TI - Alternatives to immunosuppressive drugs in human islet transplantation. AB - Although intensive insulin therapy has resulted in improved metabolic control and decreases in the incidence of complications, the occurrence of severe hypoglycemia remains an issue, as does the continued potential for complications. Islet transplantation, a promising treatment for type I diabetes, has been shown to improve blood sugar levels and decrease or even abrogate the incidence of hypoglycemia. The lack of tissue availability and the toxic effects of immunosuppressants, however, limit the application of islet transplantation as a cure for diabetes. This article discusses possible alternatives to immunosuppressive drugs in human islet transplantations. PMID- 12643200 TI - Risk factors for diabetic neuropathy and foot ulceration. AB - Peripheral sensory neuropathy (PSN) and foot ulcers are disabling and expensive complications of diabetes with few therapeutic options. Knowledge of risk factors for both complications, based on prospective observational studies, permits identification of individuals at high risk and provides options for intervention. Glycemia is the main modifiable risk factor for PSN, whereas PSN and trauma are the principal potentially modifiable risk factors for the development of foot ulceration. PMID- 12643201 TI - Measurement of somatic neuropathy for clinical practice and clinical trials. AB - Distal sensory polyneuropathy is a common and unpleasant complication of diabetes mellitus. It is the main initiating factor for foot ulceration. The increasing prevalence of diabetes has important associated health implications, both in terms of morbidity and mortality, and results in the consumption of scarce medical resources. Identification of somatic neuropathy in clinical practice is therefore important for targeted educational and other interventions. In this article, we describe methods for detecting somatic neuropathy in clinical practice and highlight those tests that are proven to be predictors of foot ulceration. The approach for detecting and characterizing somatic neuropathy for clinical trials, however, differs significantly. These methods must ideally have high levels of reproducibility, sensitivity, and specificity. Currently, several neurophysiologic tests are employed in clinical trials in order to accurately characterize diabetic neuropathy. The recent introduction of the computer assisted programs for the measurement of sensory modalities for clinical trials has been a major advance. Due to their invasive nature and associated morbidity, nerve biopsy studies are no longer used in clinical trials. Recently, using magnetic resonance imaging (MRI), significant spinal cord atrophy has been demonstrated in established neuropathy. If this observation proves to be an early feature, then a relatively rapid, noninvasive MRI technique may be used in the future to characterize diabetic neuropathy. PMID- 12643202 TI - Diagnosis and treatment of diabetic autonomic neuropathy. AB - Diabetic autonomic neuropathy (DAN) is associated with a markedly reduced quality of life and poor prognosis. The manifestations of DAN cause multiple symptoms and involve the 1) cardiovascular system: resting tachycardia, reduced heart rate variability and circadian rhythm of heart rate and blood pressure, painless myocardial ischemia/infarction, orthostatic hypotension, exercise intolerance, perioperative instability, sudden death; 2) respiratory system: reduced ventilatory drive to hypercapnia/hypoxemia, sleep apnea; 3) gastrointestinal tract: esophageal motor dysfunction, diabetic gastroparesis, gallbladder atony, diabetic enteropathy, colonic hypomotility, anorectal dysfunction; and 4) genitourinary tract: diabetic cystopathy, erectile dysfunction. Treatment is based on four cornerstones: 1) causal treatment aimed at near-normoglycemia; 2) treatment based on pathogenetic mechanisms; 3) symptomatic treatment; and 4) avoidance of risk factors and complications. Pharmacologic treatment of symptomatic DAN may be difficult, due to limited efficacy and frequent adverse reactions. First-line treatments include midodrine for orthostatic hypotension, prokinetic drugs for gastroparesis, broad-spectrum antibiotics for diabetic diarrhea, and sildenafil for erectile dysfunction. Prior to an adequate symptomatic treatment a thorough risk-benefit estimate, aimed at maintaining the patient's quality of life, is required. PMID- 12643203 TI - Update on Charcot neuroarthropathy. AB - Charcot neuroarthropathy is not uncommon in diabetic patients with peripheral neuropathy. Often, the condition is misdiagnosed for cellulitis or osteomyelitis and treatment is delayed. A high index of suspicion is required in these patients to initiate appropriate treatment early. This article covers the pathogeneses of this condition and briefly describes the recent studies performed to understand the underlying etiopathogenetic factors of this devastating condition. Lastly, it mentions the recently completed multicenter trial using bisphosphonates in diabetic Charcot neuroarthropathy. PMID- 12643204 TI - Classification of wounds of the diabetic foot. AB - A classification system should be thought of as a diagnostic language. Ideally, this language should be used by a large, diverse population of clinicians to guide therapy and ultimately predict outcome. The diabetic foot ulcer is widely known to be the prime precipitant of lower extremity amputations. Its description and classification is the first step toward widespread standardization of treatment and prevention. This article chronologically describes the major diabetic foot wound classification systems devised over the previous generation, discussing each of their attributes and their shortcomings, with an emphasis on an evidence basis for care. It is our hope that discussions such as this one will ultimately lead to a diagnostic and therapeutic lingua franca for diabetic foot wounds and a commensurate reduction in the unconscionably high prevalence of amputations, which we now currently face. PMID- 12643205 TI - The meaning of microalbuminuria in type 1 diabetes: the need for a new paradigm. PMID- 12643206 TI - Risk predictors in patients with diabetic nephropathy. AB - Diabetic nephropathy is currently the most common cause of end-stage renal disease in the Western countries. Only approximately one third of patients with type 1 diabetes develop nephropathy; thus, because it is not feasible to aggressively treat all patients, it becomes very important to find early markers in order to identify patients at high nephropathy risk. To date the best available predictor of overt nephropathy is microalbuminuria. In this article we review the validity of microalbuminuria as a predictor of overt nephropathy and consider other markers of nephropathy risk. PMID- 12643208 TI - C-peptide: a new potential in the treatment of diabetic nephropathy. AB - C-peptide is formed in the biosynthesis of insulin and the two peptides are subsequently released in equimolar amounts to the circulation. C-peptide has long been considered to be without physiologic effects. Recent data now demonstrate that C-peptide in the nanomolar concentration range binds specifically to cell surfaces, probably to G protein-coupled receptors, with subsequent activation of Ca(2+)-dependent intracellular signaling pathways and stimulation of Na+, K(+) ATPase activities. C-peptide replacement in animal models of type 1 diabetes results in diminished hyperfiltration, improved functional reserve, reduction of urinary albumin excretion, and prevention of glomerular and renal hypertrophy. Administration of C-peptide to physiologic concentrations in patients with type 1 diabetes and incipient nephropathy for periods of 3 hours to 3 months is accompanied by reduced glomerular hyperfiltration and filtration fraction, and diminished urinary albumin excretion. C-peptide replacement together with insulin therapy may be beneficial in type 1 diabetes patients with nephropathy. PMID- 12643207 TI - Treatment of hypertension in diabetic patients with nephropathy. AB - In type 1 diabetes, hypertension is closely linked to the development of nephropathy. An association of hypertension and the impact of hypertension on the clinical course of type 2 diabetes, including the development of vascular complications, has been well established. However, the association with nephropathy in type 2 diabetes is less clear. Despite that, antihypertensive treatment has a crucial impact on the course of nephropathy in both types of diabetes. In this article, we discuss recent evidence focusing on the nephroprotective potential of various classes of antihypertensive agents and confront it with current recommendations for the treatment of hypertension in diabetic patients with nephropathy. Unlike type 1 diabetes, where the nephroprotection could be a good sole measure for assessing the efficiency of a particular agent or their combination, defining of the optimal antihypertensive agent or agents in type 2 diabetes requires consideration of both cardiovascular, cerebrovascular, and nephroprotective potentials of such a treatment. In both types of diabetes, recent data support the use of inhibitors of the renin angiotensin system with or without diuretics as the initial therapy. In type 1 diabetes, additional beneficial effect can be expected from calcium channel blockers (CCBs). In type 2 diabetic patients, combining more agents may be necessary early in the course of nephropathy to affect both micro- and macrovascular targets. beta blockers should be applied early to enhance cardioprotectivity, followed by CCBs to achieve goal blood pressure. Although not supported by all recent data, aggressive blood pressure control (< 130/75 mm Hg) is warranted. Furthermore, multifactorial intervention targeting metabolic derangements and lifestyle, is a necessary complimentary measure that must accompany antihypertensive treatment. PMID- 12643209 TI - Angiotensin receptor blockers in diabetic nephropathy. AB - Where shall we place angiotensin receptor blockers in the scheme of the prevention of diabetic nephropathy? Only the results of a large, randomized double-blind trial with a comparable and appropriate alternative would prove therapeutic efficacy. The results of several trials with angiotensin-converting enzyme (ACE) inhibitors have proven them to be the standard of care for diabetics and their kidneys. As reviewed in this article, the results of three large such clinical trials have recently been completed with angiotensin receptor blockers in patients with type 2 diabetes mellitus. Initial results appear favorable. However, whether angiotensin blockers have more to offer than ACE inhibitors is still speculative. The renin-angiotensin system plays an important role in the pathogenesis of diabetic nephropathy. Since alternative pathways to ACE have been uncovered in the formation of angiotensin II, inhibition at the final end point would provide favored blockade. Because angiotensin receptor blockers do provide this specific blockade, they offer far more promise than ACE inhibitors. PMID- 12643210 TI - Genetics of diabetic nephropathy in the Pima Indians. AB - Diabetic nephropathy is the leading cause of renal failure in industrialized countries. There is strong evidence that diabetic nephropathy is influenced by genetic factors. Studies in the Pima Indians as well as in other populations demonstrate that diabetic nephropathy aggregates in families. The hypothesis that the familial aggregation reflects the effect of a major gene was formally tested by segregation analysis of diabetic nephropathy in Pima Indians with type 2 diabetes. The segregation analysis provided strong evidence for a major genetic effect on the prevalence of diabetic nephropathy; this suggests that some of the genetic determinants of diabetic nephropathy may have effects of sufficient magnitude to be detected by linkage analysis. Therefore, we analyzed data from a genome-wide scan to identify susceptibility loci for nephropathy in diabetic Pima Indians. Analyses conducted by both parametric (model-based) and nonparametric methods revealed tentative evidence for nephropathy susceptibility loci on chromosomes 3q, 7q, 18q, and 20p. PMID- 12643212 TI - [The relevance of declaring a conflict of interest in medical journals]. AB - Medical journals are often at risk of difusing research articles, reviews, position articles, editorials or letters whose message has been influenced by a conflict of interest. The readers may then be induced to accept conclusions and recommendations based on biased protocols or an unwarranted interpretation of the results. Financial support or professional links with pharmaceutical companies or other supporting agencies are the most common sources of conflict of interests, often difficult to detect. Similarly, reviews of manuscripts can be biased by personal relationships (good or bad) between reviewers and authors, by academic competition or intellectual passion, becoming other sources of conflict of interest. Even when a potential conflict of interest exists, it may not necessarily have influenced the manuscript or its review but in order to defend the transparency of the editorial process, from submission to publication, authors, reviewers and editors should declare any conflict of interest they may have and allow others to decide whether the action has been biased or not. In the present issue of Revista Medica de Chile, an updated text of the Instructions to Authors establishes that all authors should sign a statement of having or not a conflict of interest, clarifying which aspects of the work might have been affected by it. PMID- 12643211 TI - Oxidative stress in diabetic nephropathy: basic and clinical information. AB - Oxidative stress has been known to play an important role in the development and progression of diabetic nephropathy, but the intracellular signal transduction pathways regulated by reactive oxygen species (ROS) have not been clearly defined. High glucose (HG) induces intracellular ROS directly via glucose metabolism and auto-oxidation and indirectly through the formation of advanced glycation end products and their receptor binding. ROS mimic the stimulatory effects of HG and upregulate transforming growth factor-beta 1, plasminogen activator inhibitor-1, and extracellular matrix (ECM) proteins by glomerular mesangial cells, thus leading to mesangial expansion. ROS activate other signaling molecules, such as protein kinase C and mitogen-activated protein kinases and transcription factors, such as nuclear factor-kappa B, activator protein-1, and specificity protein 1 leading to transcription of genes encoding cytokines, growth factors, and ECM proteins. Finally, various antioxidants inhibit mesangial cell activation by HG and ameliorate features of diabetic nephropathy. These findings qualify ROS as intracellular messengers and as integral glucose-signaling molecules in glomerular mesangial cells in diabetic nephropathy. With this new concept, ROS assume a greater importance in the pathogenesis of diabetic nephropathy. Future studies elucidating other downstream signaling molecules activated by ROS in mesangial and other renal cells will allow us to understand the final cellular responses to HG, such as proliferation, differentiation, apoptosis, and ECM accumulation. With this new information, we should be able to develop strategies for a more rational treatment of diabetic nephropathy. PMID- 12643213 TI - [Study of polymorphism of Fc gamma IIa receptors in Chilean patients with systemic lupus erythematosus]. AB - BACKGROUND: Polymorphisms of Fc receptors for IgG (Fc gamma R) have been proposed as a genetic factor that influences susceptibility for systemic lupus erythematosus (SLE). Human Fc gamma RIIa has 2 codominantly expressed alleles, H131 and R131, which differ at amino acid position 131 in the second extracellular domain (histidine or arginine respectively) and differ substantially in their ability to bind human IgG2. The H131 allele binds IgG2 efficiently, whereas R131 binds it poorly. Because IgG2 is a poor activator of the classical complement pathway, the H131 is essential for the disposal of IgG2 immune complexes. AIM: To determine the distribution of Fc gamma RIIA genes in a cohort of Chilean SLE patients, with or without a history of lupus nephritis. PATIENTS AND METHODS: We studied 52 Chilean SLE patients fulfilling the 1982 American College of Rheumatology (ACR) criteria, 20 of whom had a history of nephritis, and 44 ethnically matched disease-free controls. Fc gamma RIIa allotypes were genotyped by PCR. RESULTS: No significant association was observed between the low affinity Fc gamma RII receptor (FcgRIIa-R131) and the presence of SLE or lupus nephritis. However, genotype frequencies in SLE patients but not in controls, departed from the proportions predicted by the Hardy-Weinberg equilibrium, suggesting this locus might be related to the disease. CONCLUSIONS: Our results suggest that in Chilean patients with SLE, as well as in many other populations, the R131 allotype is not a major factor predisposing to the development of SLE or lupus nephritis. PMID- 12643214 TI - [Digestive pathology in children infected with the human immunodeficiency virus (HIV), in Santiago de Chile]. AB - BACKGROUND: Human immunodeficiency virus (HIV) epidemiology has changed, affecting an increasing number of children. As in adults, the disease predominantly affects the digestive and respiratory systems. AIM: To report the gastrointestinal problems in HIV infected pediatric patients. PATIENTS AND METHODS: Twenty four HIV infected children (nine male, aged 1 to 12 years old, followed for 1 to 170 months), are reported. This group has been under care by a multiprofessional team. RESULTS: Oral candidiasis was present in 21 (88%), esophagic candidiasis in 3 (13%), oral ulcers in 4 (17%). Diarrhea was observed in 18 children (75%) and in eight, it had a chronic evolution. Cryptosporidium parvum was the most frequent agent found in six cases (1 with acute and 5 with chronic diarrhea). Schlerosing cholangiopathy was observed in one case, with a fatal outcome, in association to microsporidiosis. Upper endoscopy was done in 11 patients, demonstrating microscopic inflammatory changes in esophagic, gastric and duodenal epithelia in all. CONCLUSIONS: Digestive problems are common in HIV infected pediatric patients. They must be always sought actively. Endoscopy is a valuable tool for the early diagnosis of these problems. PMID- 12643215 TI - [Tissue anti-transglutaminase antibodies: a better test to detect minor dietary transgressions in celiac patients?]. AB - BACKGROUND: Endomysium antibodies (EMA) do not detect minor dietary transgressions in patients with celiac disease. AIM: To compare the sensitivity and specificity of tissue transglutaminase antibodies (tTGA) and endomysium antibodies (EMA) in biopsy proven celiac patients at the time of diagnosis and during gluten free diet (GFD). PATIENTS AND METHODS: One hundred fifty three subjects were studied: a) 30 healthy controls; b) 9 cases with cow's milk allergy; c) 24 celiac patients at time of diagnosis; d) 25 celiac patients adhering to the GFD; e) 65 celiac patients with poor/no adhesion to GFD. EMA and tTGA IgA were measured by immunofluorescence and ELISA, respectively. RESULTS: Sensitivity and specificity were 100% and 97.4% for tTGA, respectively. All patients with cow's milk allergy were EMA (-) and 8 of 9 (88.9%) were tTGA (-). In celiac patients not adhering to the GFD, EMA and tTGA positivity were similar (80% and 81.5%, respectively); 95.4% of the subjects tested positive for at least one of them. All patients adhering to GFD were EMA (-) but tTGA were (+) in 28% of them. CONCLUSIONS: EMA and tTGA have similar sensitivity and specificity at the time of diagnosis of celiac disease. Positive tTGA in 28% of patients that adhered strictly to the GFD and whose EMA were negative suggest that tTGA may be helpful in detecting minor dietary transgressions and should be further evaluated. PMID- 12643216 TI - [Antitransglutaminase antibodies determination for the diagnosis of celiac disease]. AB - BACKGROUND: The diagnosis of celiac disease is based in clinical features, serology and intestinal biopsy. There are recent reports that antiglutaminase antibodies have a good correlation with anti endomisial antibodies. AIM: To assess the sensitivity and specificity of antitransglutaminase antibodies in the diagnosis of celiac disease and their correlation with antiendomisial antibodies. MATERIAL AND METHODS: Forty nine patients with celiac disease (mean age 30 years, 12 male) were studied. Thirty were symptomatic and 19, asymptomatic. As controls, 34 subjects (mean age 27 years, seven male), with normal duodenal biopsies, were studied. Sera was processed for the determination of antigliadin IgA by ELISA, antiendomisium IgA by indirect immunofluorescence, and antitransglutaminase IgA by ELISA. RESULTS: Antigliadin antibodies had a sensitivity of 73%, a specificity of 96%, a positive predictive value of 93% and a negative predictive value of 82% for the diagnosis of celiac disease. Antiendomisium and antitransglutaminase antibodies had a specificity and positive predictive value of 100%, sensitivities of 89 and 92% respectively and negative predictive values of 92 and 94% respectively. No significant differences in the diagnostic yield of antiendomisium and antitransglutaminase antibodies, were observed. CONCLUSIONS: In this group of patients, antitransglutaminase antibodies had a high concordance with antiendomisium antibodies, for the diagnosis of celiac disease. Considering that the determination of antitransglutaminase antibodies is of lower cost and less complicated than antiendomisium antibodies, it is a useful tool for the diagnosis and follow up of patients with celiac disease. PMID- 12643217 TI - [Cholelithiasis in children: a clinical and morphological study]. AB - BACKGROUND: The use of ultrasonography increased the frequency of diagnosis of cholelithiasis in childhood. AIM: To determine the clinical and laboratory features and follow up of children with biliary stones. PATIENTS AND METHODS: Twenty six children (13 male, aged 1 month to 14 years) were prospectively enrolled. RESULTS: Nine children had a past medical history of factors potentially predisposing to stones. A clinical presentation with vomiting (50%), abdominal pain (46%) and jaundice (23%) was the most common indication for surgery. The diagnosis was based on abdominal ultrasound in all children. Cholecystectomy was performed in 15 children (laparoscopy in 13 and open surgery in 2). Children who underwent surgery were older than those who did not undergo surgery (p < 0.001), but they did not have differences in liver function tests. Eight children had pigmented stones and seven had cholesterol stones. CONCLUSIONS: In our patients, neither family history nor laboratory tests were useful in the diagnosis as well as in the clinical decision of surgery, which was based on symptoms. The presence of cholesterol stones in a high proportion of these children may be a unique situation in Chile, considering the high prevalence of this disease in the adult population. PMID- 12643218 TI - [Abnormalities of adrenal steroidogenesis in Chilean boys with micropenis]. AB - BACKGROUND: Mutations in type II 3 beta hydroxysteroid Dehydrogenase (3 beta HSD) are found in male children with severe undervirilized genitalia. Mild undervirilization (isolated micropenis or with distal hypospadia) can be associated with a partial deficit in 3 beta HSD activity. AIM: To assess the frequency of abnormal adrenal response to ACTH, suggesting a deficit in adrenal enzymatic activity, in children with mild undervirilization. PATIENTS AND METHODS: We studied 26 male children with micropenis, aged one to eight years. Children with evidences of puberal development or in treatment with drugs that affect steroidal metabolism were excluded from the study. Serum levels of androstenedione (A), dehydroepiandrosterone (DHEA), progesterone (P), 17 hydroxyprogesterone (17 P) and the ratios DHEA/A, P/17 P, 17 P/DHEA were measured after an adrenal stimulation with 0.25 mg/m2 intramuscular ACTH. RESULTS: Two children had DHEA y DHEA/A values suggesting a defective 3 beta HSD activity. Other two children had high levels of 17 P, suggesting a deficiency of cytochrome p450c21. A CYP 21 gene mutation was found in one of the later children. CONCLUSIONS: A low proportion of children with micropenis have a deficient 3 beta HSD activity. PMID- 12643219 TI - [Reproductive risk of women over 40 years old]. AB - BACKGROUND: Pregnancy at extreme ages is a risk factor for maternal, perinatal and infantile death. AIM: To assess the obstetric risk of women over 40 years old. MATERIAL AND METHODS: The risk of maternal, late fetal, neonatal and infantile deaths and the risk of low birth weight was assessed in women over 40 years old and compared to the risk of women aged 20-34 years. Data was obtained from the 1999 Annual Report of the National Statistics Institute and the odds ratios (OR) with a 95% confidence interval were calculated. RESULTS: Women over 40 years had a higher risk of maternal death (OR 7.13, 3.31-14.97), late fetal death (OR 2.19, 1.69-2.85), neonatal death (OR 1.8, 1.4-2.32), infantile death (OR 1.8, 1.49-2.18) and low birth weight (OR 1.72, 1.58-1.88). CONCLUSIONS: The higher reproductive risk of women over 40 years is confirmed. Better birth control programs at this age will reduce this risk. PMID- 12643220 TI - [A retrospective study of 232 cases of lip cancer and pre cancer in Chilean patients. Clinical-histological correlation]. AB - BACKGROUND: The most common type of oral cancer is squamous cell carcinoma, which accounts for approximately 90% of all oral malignancies. Therefore the oral cancer problem primarily concerns the diagnosis, biology and management of squamous cell carcinoma (SCC). However, there are red and white lesions, that are considered premalignant, since they may unpredictably transform into cancer. According to WHO, leukoplakia, eritroplakia and actinic cheilitis are considered precancerous lesions which pathologically may correspond to intraepithelial dysplasia, hyperkeratosis, carcinoma in situ or SCC. AIM: To study the clinical and pathological features of lip cancer and premalignant lip lesions. PATIENTS AND METHODS: A retrospective analysis of 232 cases with lip lesions. The frequency, location, gender, age and pathological diagnosis of the lesions were recorded. RESULTS: One hundred fifty four lesions were observed in men and 78 in women, 92% of lesions were located in the lower lip. Seventy two of the 232 lesions were SCC and 72% of these carcinomas were well differentiated. The age of patients with cancer was higher than that of patients with precancerous lesions. CONCLUSIONS: The difference in age between patients with cancer and precancerous lesions, reinforces the existing knowledge that precancerous lesions may transform into cancer if they are not properly diagnosed and treated. PMID- 12643221 TI - [Serologic evidence for human Ehrlichiosis in Chile]. AB - BACKGROUND: Ehrlichiosis is a non contagious infectious disease, mainly transmitted by thick bites. In 1998, this infection was detected in dogs, for the first time, in Chile. AIM: To establish if there is human exposure to Ehrlichia sp in Chile. MATERIAL AND METHODS: Blood samples from 17 dogs with ehrlichiosis and 19 humans who had contact with them were studied to determine human exposure to Ehrlichia equi and Ehrlichia chaffeensis in Chile. Samples were analyzed by indirect immunofluorecence and by polymerase chain reaction (PCR). RESULTS: Six dogs had positive titers against both species of ehrlichia; 2 with titers of 1/256; 3 with titers over 1/512 to Ehrlichia equi and titers of 1/256, 1/128 and 1/64 to Ehrlichia chaffeensis respectively, and 1 with titers of 1/256 to Ehrlichia equi and titers of 1/128 to Ehrlichia chaffeensis. Two of the 19 humans, had positive titers against both antigens (1/128). PCR reactions were negative in both human and canine sera. CONCLUSIONS: These results confirm that human exposure to Ehrlichia sp. Epidemiological surveillance for human ehrlichiosis should be implemented in the country. PMID- 12643222 TI - [Peripheral precocious puberty caused by a juvenile granulosa cell ovarian tumor, with iso and heterosexual manifestations in a six years old girl]. AB - A six years old girl consulted due to mammary development. On physical examination, clitoris enlargement and a tumor localized in the abdominal-pelvic region were observed. Hormonal study disclosed elevated testosterone and estradiol levels. On exploratory laparotomy, a right ovarian tumor was observed and a right salpingooophorectomy was performed. The contemporary biopsy informed a disgerminoma, leading to a surgical staging of the tumor. The definitive pathological diagnosis was a juvenile granular cell tumor, limited to the ovary. In the postoperative period, estradiol and testosterone levels returned to normal values and the pseudopuberty reverted. The patient did not receive adjuvant treatment and after three years of follow up, there is no evidence of tumor recidivism. PMID- 12643223 TI - [Paecilomyces lilacinus systemic infection in an immunocompromised child]. AB - The incidence of systemic fungal infections increased during the last two decades. Rare fungi, such as Mucor, Fusarium and Paecilomyces, are emerging as causes of systemic fungal infections in immunocompromised hosts. There are reports of cutaneous infections, endophthalmitis, keratitis, sinusitis, neuropathy and fungemia in immunocompromised and immunocompetent adult patients. We report a 5 years old neutropenic patient with acute myeloid leukemia treated with multiple courses of chemotherapy, with a fungemia caused by Paecilomyces lilacinus (PL). His initial clinical course was characterized by fever, skin lesions, respiratory distress and shock. Blood and bone marrow cultures were positive. The patient was treated with amphotericin B and itraconazole with a good clinical response. PMID- 12643224 TI - [Eosinophilic pneumonia caused by mesalazine. Report of one case]. AB - A 50 years old male with a diagnosis of ulcerative colitis treated with mesalazine, developed after 2 months of treatment, cough, fever and progressive dyspnea. Chest X ray examination and CT scan showed pulmonary infiltrates in the right upper lobe that subsequently involved both lower lobes. A biopsy, made through videothoracoscopy, showed an eosinophilic pneumonia. After the discontinuation of mesalazine and the use of glucocorticoids, the respiratory involvement resolved, and pulmonary infiltrates regressed. Mesalazine is widely used in the treatment of inflammatory bowel diseases. Pulmonary toxicity is an uncommon complication of mesalazine treatment. Nevertheless, this complication should be considered in patients that use it and develop respiratory symptoms. PMID- 12643225 TI - [Visceral hypersensitivity: a concept within our reach]. AB - Despite significant advances in the recognition of etiological factors and pathological mechanisms, the pathophysiology of functional gastrointestinal disorders (FGD) is still not fully understood. Visceral hypersensitivity has been recognized as a characteristic of patients with FGD, especially in patients with irritable bowel syndrome (IBS). Visceral afferent input is modulated by a variety of mechanisms, operating between the gastrointestinal tract and the brain. Dysfunction of these regulatory mechanisms could distort gastrointestinal perceptions. Recent findings suggest that in the majority of cases of IBS the primary abnormality may be at the periphery with alterations of the motor and secretory sensory activity. Although imaging techniques indicate that there are also differences in cortical activation. Furthermore, selective serotonin reuptake inhibitors may benefit FGD. Recent pharmacological studies suggest that 5-HT3 antagonist such as alosetron and cilansetron, and 5-HT4 agonist such as legaserod and prucalopride may also have a potential use in FGD. PMID- 12643226 TI - [Conflict of interest in biomedical journals]. PMID- 12643227 TI - [Consensus on the diagnosis and treatment of central early puberty]. AB - The beginning of puberty is marked by breast growth in girls and testicular enlargement in boys. These occur at the age of 10.5 +/- 2.0 years in females and 11.5 +/- 2.0 years in males. Recent but controversial publications suggest that these events are being observed at younger ages, at least in the USA. There are no studies demonstrating that this is true in Chile. For this reason we still consider that puberty is precocious when it occurs before 8.0 years in girls and before 9.0 years in boys. True or central precocious puberty (CPP) must be distinguished from peripheral or pseudoprecocious puberty (PPP), from premature telarche and from premature adrenarche. We suggest that the workup of a patient with premature development should include an LHRH test to demonstrate if the hypothalamic-pituitary axis is activated, plasma levels of sex steroids, bone age and pelvic ultrasound in girls. All children with CPP should have a CAT scan or MNR of the brain, since a lesion of the central nervous system is observed in 15% of the girls and 50% of the boys with CPP. Additional studies are needed in cases of PPP. The aim of treating CPP is to avoid adult short stature that results from premature fusion of the epiphysis and to avoid eventual emotional and psychological stress. Treatment consists of monthly intramuscular injections of a depot preparation of LHRH analogs. Suppression of pituitary and gonadal activity produces regression of secondary sex characteristics and slowing down of growth velocity and bone maturation. The opportunity, duration of treatment and their effect on final stature are discussed. PMID- 12643228 TI - [On the ownership of Medical Records]. AB - It is not clear if medical records belong to the physician, the institution or the patient. There is a bill being discussed in the Chilean Congress that establishes that "the patient, personally or through a representative, has the right to access and review his medical record. In case of death, this right may be exerted by his inheritors". In this article we postulate that this bill infringes a number of legal norms in force and universally accepted ethical principles. We distinguish between patient's advocacy to be informed and their free access to medical records. The main ethical principles violated are those of beneficence and non maleficence. PMID- 12643229 TI - The importance of number of blastomeres when embryos are transferred in the absence of controlled ovarian hyperstimulation. AB - PURPOSE: To determine if the previous findings that transferring embryos with a higher number of blastomeres results in higher pregnancy rates following fresh but not frozen embryo transfer (ET) was related to the use of controlled ovarian hyperstimulation (COH) in the former but not in the latter. METHODS: Retrospective review of pregnancy and implantation rates following fresh embryo transfer of donor egg recipient cycles (where no COH is used) vs frozen ETs during the same time period according to whether there was at least one embryo with eight blastomeres transferred or not. RESULTS: Significantly higher pregnancy rates with an 8-cell ET in donor oocyte recipient cycles but not frozen ETs. CONCLUSIONS: A less favorable uterine environment caused by the use of high dose gonadotropin is not responsible for the once again observed difference in higher pregnancy rates with higher blastomere number in fresh vs frozen ET. However, an effect of the gonadotropin releasing hormone analogue was not ruled out by this study. PMID- 12643230 TI - Methicillin-resistant Staphylococcus aureus in an adult military beneficiary population lacking risk factors: susceptibility to orally available agents. AB - To determine the unique susceptibility of community-acquired methicillin resistant Staphylococcus aureus (MRSA) as a function of the presence or absence of risk factors in a military beneficiary population, we reviewed all MRSA cases between 1994 and 1997 in a military medical center. Of 67 cases, 24 were community acquired, 9 of whom lacked risk factors for MRSA infection, and 43 cases were nosocomial. Among isolates from patients without risk factors, seven (77.8%) were susceptible to ciprofloxacin, eight (88.9%) to trimethoprim/sulfamethoxazole, eight (88.9%) to erythromycin, eight (88.9%) to tetracycline, and nine (100.0%) to clindamycin. These rates were markedly higher than those seen in isolates obtained from community-acquired cases with risk factors as well as from nosocomial cases (p < 0.01). Three clonal types of MRSA from patients without risk factors were identified and susceptibilities were similar for each of the three types. These results indicate that therapy with active oral agents might be reasonable for some less severe MRSA infections in our active duty military patients without risk factors, and additional studies are merited. PMID- 12643232 TI - Recent changes to FDA--approved labeling. PMID- 12643231 TI - New drugs and dosage forms. PMID- 12643234 TI - Ophthalmo-acromelic syndrome in a Turkish infant: case report. AB - Ophthalmo-acromelic syndrome (OAS) is an extremely rare autosomal recessive disorder characterised by eye malformations ranging from true anophthalmia to mild microophthalmia and acromelic malformations. In this article, we report a newborn infant with OAS because of its rare presentation. He was the fourth sibling affected in the family. The parents were healthy but there was a close blood relationship between the parents. Physical examination revealed bilateral true anophthalmia and oligodactyly (bilateral four toes) on the feet. He had no other additional abnormalities. We consider that this rare syndrome could be relatively more common in our country because six Turkish cases of OAS have been reported in the English literature to date. PMID- 12643233 TI - Mycoplasma pneumoniae in children with pneumonia at Mbagathi District Hospital, Nairobi. AB - OBJECTIVE: To determine the prevalence of mycoplasma pneumoniae in nasopharyngeal aspirates of children under five years of age suffering from pneumonia. DESIGN: Cross-sectional survey. METHODS: Two primer sets targeting the genes coding for adhesion protein and 16S rRNA were used in PCR and M. pneumoniae specific antibodies were detected using commercial article agglutination kit. Microbiological investigations to isolate common acute respiratory infectious pathogens were also carried out. RESULTS: M. pneumoniae was detected in nasopharyngeal aspirates of 33.7% of the cases by PCR while serology was positive in only 4.1%. M. pneumoniae alone (single detection) was detected in 25% of the cases while 75% occurred with other acute respiratory infectious (ARI) pathogens. However, the results did not correlate with clinical outcome or the severity of pneumonia. No significant aetiology was found in 28% of the cases investigated, however microbiological investigations by culture revealed the presence of other aetiological agents as follows: Streptococcus pneumoniae (26%), Klebsiella pneumoniae (1%), Staphylococcus aureus (3%), E. coli (2%), parainfluenza viruses (5%), Salmonella(1%), adenovirus (4%), RSV (22%) and Candida spp(13%), Mycoplasma pneumoniae was more prevalent in children aged between six months and three years. Cases of M. pneumoniae PCR positive and S. pneumoniae exhibited similar seasonal distribution with peaks in May and September. However, there was no relationship between M. pneumoniae PCR positive and the severity of pneumonia. CONCLUSION: More investigation is required to establish the significance of atypical pathogens in respiratory infections in Kenya. PMID- 12643235 TI - High-contrast fast Fourier transform acousto-optical tomography of phantom tissues with a frequency-chirp modulation of the ultrasound. AB - We report new results on acousto-optical tomography in phantom tissues using a frequency chirp modulation and a CCD camera. This technique allows quick recording of three-dimensional images of the optical contrast with a two dimensional scan of the ultrasound source in a plane perpendicular to the ultrasonic path. The entire optical contrast along the ultrasonic path is concurrently obtained from the capture of a film sequence at a rate of 200 Hz. This technique reduces the acquisition time, and it enhances the axial resolution and thus the contrast, which are usually poor owing to the large volume of interaction of the ultrasound perturbation. PMID- 12643237 TI - Cases from the Osler Medical Service at Johns Hopkins University. PMID- 12643238 TI - Abstracts of the annual meeting of the Association of Academic Physiatrists. Fort Lauderdale, Florida, USA. March 27-29, 2003. PMID- 12643236 TI - Colpoplasty and acute pneumonitis. PMID- 12643239 TI - [Vaccination. Tasks and obligations for surgeons]. PMID- 12643240 TI - [Comment on the survey of GMK organization: "Threatened physician shortage in Germany"]. PMID- 12643241 TI - [Future prospects for young surgeons. German Society of Surgery memorandum]. PMID- 12643242 TI - [Declaration by physician and nursing staff of the Neumunster Friedrich-Ebert hospital]. PMID- 12643243 TI - [1. National DRG Forum--a forum for getting answers to immediately relevant questions]. PMID- 12643244 TI - [Clinical treatment pathways: successful due to standardization. Better cooperation, clear responsibilities, cost overview and better quality]. PMID- 12643245 TI - [G-DRG version 1.0--it becomes a serious issue now]. PMID- 12643246 TI - [Life-saving amputation. Reform policy for Berlin university clinics demonstrates the future path of German university medicine: less patient care, more specialization--and perhaps also research]. PMID- 12643249 TI - [High noon in public health politics?]. PMID- 12643251 TI - [Report on the 16th Surgery Meeting Day]. PMID- 12643252 TI - Abstracts of the Spring Meeting of the WPSA UK Branch. York, United Kingdom. April 9-10, 2002. PMID- 12643253 TI - [Clinical research in Germany. The problems involved in patient-oriented research and the conducting of clinical studies in German university hospitals]. PMID- 12643254 TI - Developmental toxicity of pyrethroids. PMID- 12643256 TI - [Reimbursement questions for cardiology services within the scope of GOA:2. Position of the DRG and GOA Project Group]. PMID- 12643257 TI - Study finds stroke symptoms differ between the sexes. PMID- 12643258 TI - Mortality rates are higher in Penn. hospitals with lower nurse staffing, study finds. PMID- 12643259 TI - Reducing eye pressure slows glaucoma progression. PMID- 12643260 TI - Circulating ghrelin concentrations are reduced in healthy offspring of Type 2 diabetic subjects, and are increased in women independent of a family history of Type 2 diabetes. PMID- 12643261 TI - Nurse practitioners improve risk factor management for patients with coronary heart disease. PMID- 12643262 TI - E23K single nucleotide polymorphism in the islet ATP-sensitive potassium channel gene (Kir6.2) contributes as much to the risk of Type II diabetes in Caucasians as the PPARgamma Pro12Ala variant. PMID- 12643263 TI - Replication-independent MCB gene induction and deoxyribonucleotide accumulation at G1/S in Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, many genes encoding enzymes involved in deoxyribonucleotide synthesis are expressed preferentially near the G1/S boundary of the cell cycle. The relationship between the induction of deoxyribonucleotide synthesizing genes, deoxyribonucleoside triphosphate levels, and replication initiation was investigated using factor-synchronized wild-type yeast or dbf4 yeast that are temperature-sensitive for replication initiation. Neither the timing nor extent of gene induction was inhibited when factor-arrested dbf4 cells were released into medium containing the ribonucleotide reductase inhibitor hydroxyurea, which blocks replication fork progression, or were released at 37 degrees C, which blocks replication origin firing. Thus, the induction of deoxyribonucleotide-synthesizing genes at G1/S was fully independent of DNA chain elongation or initiation. Deoxyribonucleoside triphosphate levels increased severalfold at G1/S in wild-type cells and in dbf4 mutants incubated at the non permissive temperature. Thus, deoxyribonucleoside triphosphate accumulation, like the induction of deoxyribonucleotide-synthesizing genes, was not dependent on replication initiation. Deoxyribonucleoside triphosphate accumulation at G1/S was suppressed in cells lacking Swi6, a transcription factor required for normal cell cycle regulation of deoxyribonucleotide-synthesizing genes. The results suggest that cells use gene induction at G1/S as a mechanism to pre-emptively, rather than reflexively, increase the synthesis of DNA precursors to meet the demand of the replication forks for deoxyribonucleotides. PMID- 12643266 TI - Abstracts of the International Conference on Tumor Cell Dissemination in Breast Cancer. 24-25 January 2003, Heidelberg, Germany. PMID- 12643264 TI - Calcium induces cell survival and proliferation through the activation of the MAPK pathway in a human hormone-dependent leukemia cell line, TF-1. AB - Survival and proliferation of cells of a human myelo-erythroid CD34+ leukemia cell line (TF-1) depend on the presence of granulocyte-macrophage colony stimulating factor or interleukin-3. Upon hormone withdrawal these cells stop proliferating and undergo apoptotic process. In this report we demonstrate that a controlled increase in [Ca2+]i induces hormone-independent survival and proliferation of TF-1 cells. We found that moderate elevation of [Ca2+]i by the addition of cyclopiasonic-acid protected TF1 cells from apoptosis. Furthermore, a higher, but transient elevation of [Ca2+]i by ionomycin treatment induced cell proliferation. In both cases caspase-3 activity was reduced, and Bcl-2 was up regulated. Higher elevation of [Ca2+]i by ionomycin induced MEK-dependent biphasic ERK1/2 activation, sufficient to move the cells from G0/G1 to S/M phases. Meanwhile, activation of ERK1/2, phosphorylation of the Elk-1 transcription factor, and, consequently, a substantial elevation of Egr-1 and c Fos levels and AP-1 DNA binding were observed. Moderate elevation of [Ca2+]i, on the other hand, caused a delayed monophasic activation of ERK1/2 and Elk-1 that was accompanied with only a small increase of Egr-1 and c-Fos levels and AP-1 DNA binding. The specific MEK-1 kinase inhibitor, PD98059, inhibited all the effects of increasing [Ca2+]i, indicating that the MAPK/ERK pathway activation is essential for TF-1 cell survival and proliferation. Based on these results we suggest that the elevation of the [Ca2+]i may influence the cytokine dependence of hemopoietic progenitors and may contribute to pathological hematopoiesis. PMID- 12643267 TI - Abstracts of the 52nd Annual Scientific Session of the American College of Cardiology. Chicago, Illinois, USA. March 30-April 2, 2003. PMID- 12643265 TI - Localisation of congenital glioblastomas in the Japanese. PMID- 12643268 TI - Reversed circadian rhythms of heart rate variability and morning peak occurrence of sustained ventricular tachyarrhythmias in patients with implanted cardioverter defibrillator. PMID- 12643269 TI - Abstracts of the American Urological Association Annual Meeting. Chicago, Illinois, USA. April 26-May 1, 2003. PMID- 12643270 TI - Evaluation of routine imaging after ventriculoperitoneal shunt insertion. PMID- 12643271 TI - A tribute to James E. Bailey. PMID- 12643272 TI - Retraction. Cloning and expression of the AtGRP9 gene related to salt stress tolerance in Arabidopsis thaliana. PMID- 12643273 TI - [VII Educational Conference. Obstruction of respiratory airways--new drugs for allergies, asthma and COPD. Jachranka, 29 August-1 September 2002. Proceedings and abstracts]. PMID- 12643275 TI - [High frequency surgery in arthroscopy]. PMID- 12643274 TI - Signaling of human ciliary neurotrophic factor (CNTF) revisited. The interleukin 6 receptor can serve as an alpha-receptor for CTNF. AB - Human ciliary neurotrophic factor (CNTF) is a neurotrophic cytokine that exerts a neuroprotective effect in multiple sclerosis and amyotrophic lateral sclerosis. Clinical application of human CNTF, however, was prevented by high toxicity at higher dosages. Human CNTF elicits cellular responses by induction of a receptor complex consisting of the CNTF alpha-receptor (CNTFR), which is not involved in signal transduction, and the beta-receptors gp130 and leukemia inhibitory factor receptor (LIFR). Previous studies with rat CNTF demonstrated that rat CNTF is unable to interact with the human interleukin-6 alpha-receptor, whereas at high concentrations, it can directly induce a signaling heterodimer of human gp130 and human LIFR in the absence of the CNTF receptor. Here, we demonstrate that human CNTF cannot directly induce a heterodimer of human gp130 and LIFR. However, human CNTF can use both the membrane-bound and the soluble human IL-6R as a substitute for its cognate alpha-receptor and thus widen the target spectrum of human CNTF. Engineering a CNTFR-specific human CNTF variant may therefore be a prerequisite to improving the safety profile of CNTF. PMID- 12643276 TI - Stabilization versus inhibition of TAFIa by competitive inhibitors in vitro. AB - Two competitive inhibitors of TAFIa (activated thrombin-activable fibrinolysis inhibitor), 2-guanidinoethylmercaptosuccinic acid and potato tuber carboxypeptidase inhibitor, variably affect fibrinolysis of clotted human plasma. Depending on their concentration, the inhibitors shortened, prolonged, or had no effect on lysis in vitro. The inhibitor-induced effects were both tissue-type plasminogen activator (tPA) and TAFIa concentration-dependent. Inhibitor dependent prolongation was favored at lower tPA concentrations. The magnitude of the prolongation increased with TAFIa concentration, and the maximal prolongation observed at each TAFIa concentration increased saturably with respect to TAFIa. A theoretical maximal prolongation of 20-fold was derived from a plot of the maximum prolongation versus TAFIa. This represents, for the first time, a measurement of the maximal antifibrinolytic potential of TAFIa in vitro. Because TAFIa spontaneously decays, the stabilization of TAFIa was investigated as a mechanism explaining the inhibitor-dependent prolongation of lysis. Both inhibitors stabilized TAFIa in a concentration-dependent, non-saturable manner. Although their KI values differed by three orders of magnitude, TAFIa was identically stabilized when the fraction of inhibitor-bound TAFIa was the same. The data fit a model whereby only free TAFIa decays. Therefore, the variable effects of competitive inhibitors of TAFIa on fibrinolysis can be rationalized in terms of free TAFIa and lysis time relative to the half-life of TAFIa. PMID- 12643277 TI - A new perspective on V3 phenotype prediction. AB - The particular coreceptor used by a strain of HIV-1 to enter a host cell is highly indicative of its pathology. HIV-1 coreceptor usage is primarily determined by the amino add sequences of the V3 loop region of the viral envelope glycoprotein. The canonical approach to sequence-based prediction of coreceptor usage was derived via statistical analysis of a less reliable and significantly smaller data set than is presently available. We aimed to produce a superior phenotypic classifier by applying modern machine learning (ML) techniques to the current database of V3 loop sequences with known phenotype. The trained classifiers along with the sequence data are available for public use at the supplementary website: http://genomiac2.ucsd.edu:8080/wetcat/v3.html and http://www.cs.waikato.ac.nz/ml/weka[corrected]. PMID- 12643278 TI - Stimulated interaction between and subunits of tryptophan synthase from hyperthermophile enhances its thermal stability. AB - Tryptophan synthase from hyperthermophile, Pyrococcus furiosus, was found to be a tetrameric form (22) composed of and 2 subunits. To elucidate the relationship between the features of the subunit association and the thermal stability of the tryptophan synthase, the subunit association and thermal stability were examined by isothermal titration calorimetry and differential scanning calorimetry, respectively, in comparison with those of the counterpart from Escherichia coli. The association constants between the and subunits in the hyperthermophile protein were of the order of 108 M1, which were higher by two orders of magnitude than those in the mesophile one. The negative values of the heat capacity change and enthalpy change upon the subunit association were much lower in the hyperthermophile protein than in the mesophile one, indicating that the conformational change of the hyperthermophile protein coupled to the subunit association is slight. The denaturation temperature of the subunit from the hyperthermophile was enhanced by 17 degrees C due to the formation of the 22 complex. This increment in denaturation temperature due to complex formation could be quantitatively estimated by the increase in the association constant compared with that of the counterpart from E. coli. PMID- 12643279 TI - Prevalence of drug-resistance-associated mutations in antiretroviral drug-naive Zambians infected with subtype C HIV-1. AB - The ability of HIV-1 to evolve resistance to antiretroviral drugs leads to treatment failure. By nucleotide sequencing of HIV-1 subtype B isolates, amino acids responsible for drug resistance have been identified. Less information is available, however, on the extent and distribution of these amino acids in HIV-1 nonsubtype B viruses circulating mainly in developing countries. More HIV infected patients in the developing world are now using antiretroviral drugs, and hence there is a need to monitor drug resistance mutations in HIV-1 non-subtype B viruses. This study examines the prevalence of drug resistance mutations in 28 antiretroviral drug-naive HIV-1-infected Zambians. HIV-1 proviral DNA was extracted from peripheral blood mononuclear cells. The region encompassing gag p17 to env C2-V3-C3 was amplified by the polymerase chain reaction followed by direct sequencing. Sequence analyses for drug resistance-associated mutations in th e protease and reverse transcriptase genes, and HIV-1 subtyping, were done. Overall, 92.8% of the generated sequences were HIV-1 subtype C. The generated sequences revealed only secondary associated, but no primary, drug-resistance mutations The most frequent secondary mutations in the protease and RT genes were, respectively, I93L(91.7%), L89M (79.2%), M3611V (79%, 4.2%), and R211K (70.8%), S48T (62.5%). The atypical residues M41N (3.6%) and D67A (3.6%) were detected in the RT gene. This study reveals many naturally occurring polymorphisms in HIV-1 subtype C isolates from antiretroviral drug-naive individuals. Such polymorphisms could lead to rapid treatment failure and development of drug-resistant HIV-1 mutants in individuals undergoing antiretroviral therapy. PMID- 12643280 TI - A polymorphism in thrombospondin-1 associated with familial premature coronary heart disease causes a local change in conformation of the Ca2+-binding repeats. AB - A single nucleotide polymorphism that substitutes a serine for an asparagine at residue 700 in the Ca2+-binding repeats of thrombospondin-1 is associated with familial premature coronary heart disease. We expressed the Ca2+-binding repeats alone (Ca) or with the third epidermal growth factor-like module (E3Ca), without (Asn-700) or with (Ser-700) the disease-associated polymorphism. The intrinsic fluorescence of a single tryptophan (Trp-698) adjacent to the polymorphic residue was quenched cooperatively by adding Ca2+. The third epidermal growth factor-like repeat dramatically altered the Ca2+-dependent fluorescence transition for the Asn-700 constructs; the half-effective concentration (EC50) of Ca Asn-700 was 390 microM, and the EC50 of E3Ca Asn-700 was 70 microM. The Ser-700 polymorphism shifted the EC50 to higher Ca2+ concentrations (Ca Ser-700 EC50 of 950 microM and E3Ca Ser-700 EC50 of 110 microM). This destabilizing effect is due to local conformational changes, as the Ser-700 polymorphism did not influence the secondary structure of E3Ca or Ca as assessed by far UV circular dichroism. At 200 microM Ca2+, in which both E3Ca Asn-700 and Ser-700 are in the Ca2+-replete conformation at 37 degrees C, the fluorescence of E3Ca Ser-700 reverted to the Ca2+-depleted spectrum at 50 degrees C compared with 65 degrees C for E3Ca Asn 700. These findings indicate that the Ser-700 polymorphism subtly but significantly sensitizes the calcium-binding repeats to removal of Ca2+ and thermal denaturation. PMID- 12643281 TI - Identification of env CRF-10 among HIV variants circulating in rural western Kenya. AB - As part of a program to determine the genetic diversity of human immunodeficiency virus in rural Kenya, we carried out a molecular analysis of the C2-V3 region of HIV-infected blood samples obtained from 30 antenatal clinic attendees of seven health centers in western Kenya. Direct sequencing was carried out on the envelope C2-V3 region of proviral DNA. On phylogenetic analysis with reference strains, 20 were subtype Al, 2 were subtype D, 1 was subtype C, 1 was subtype G, 1 was CRF-10, 2A/D, 2A/C, and 2 were unclassified. The presence of CRF-10 and the great variety of subtypes and recombinants in such a limited sample size suggest that western Kenya may be a potential hotspot for HIV recombination in the country. PMID- 12643283 TI - Rad23 ubiquitin-associated domains (UBA) inhibit 26 S proteasome-catalyzed proteolysis by sequestering lysine 48-linked polyubiquitin chains. AB - Most substrates of the 26 S proteasome are recognized only following conjugation to a Lys48-linked polyubiquitin chain. Rad23 is one member of a family of proteins that possesses an N-terminal ubiquitin-like domain (UbL) and a C terminal ubiquitin-associated domain(s) (UBA). Recent studies have shown that UbLs interact with 26 S proteasomes, whereas UBAs bind polyubiquitin chains. These biochemical properties suggest that UbL-UBA proteins may shuttle polyubiquitinated substrates to proteasomes. Here we show that contrary to prediction from this model, the effect of human Rad23A on the degradation of polyubiquitinated substrates catalyzed by purified proteasomes is exclusively inhibitory. Strong inhibition is dependent on the presence of both UBAs, independent of the UbL, and can be explained by competition between the UBA domains and the proteasome for binding to substrate-linked polyubiquitin chains. The UBA domains bind Lys48-linked polyubiquitin chains in strong preference to Lys63 or Lys29-linked chains, leading to selective inhibition of the assembly and disassembly of Lys48-linked chains. These results place constraints on the mechanism(s) by which UbL-UBA proteins promote proteasome-catalyzed proteolysis and reveal new properties of UBA domains. PMID- 12643282 TI - Eosinophil peroxidase-derived reactive brominating species target the vinyl ether bond of plasmalogens generating a novel chemoattractant, alpha-bromo fatty aldehyde. AB - Plasmalogens are a subclass of glycerophospholipids that are enriched in the plasma membrane of many mammalian cells. The vinyl ether bond of plasmalogens renders them susceptible to oxidation. Accordingly, it was hypothesized that reactive brominating species, a unique oxidant formed at the sites of eosinophil activation, such as in asthma, might selectively target plasmalogens for oxidation. Here we show that reactive brominating species produced by the eosinophil peroxidase system of activated eosinophils attack the vinyl ether bond of plasmalogens. Reactive brominating species produced by eosinophil peroxidase target the vinyl ether bond of plasmalogens resulting in the production of a neutral lipid and lysophosphatidylcholine. Chromatographic and mass spectrometric analyses of this neutral lipid demonstrated that it was 2-bromohexadecanal (2 BrHDA). Reactive brominating species produced by eosinophil peroxidase attacked the plasmalogen vinyl ether bond at acidic pH. Bromide was the preferred substrate for eosinophil peroxidase, and chloride was not appreciably used even at a 1000-fold molar excess. Furthermore, 2-BrHDA production elicited by eosinophil peroxidase-derived reactive brominating species in the presence of 100 microM NaBr doubled with the addition of 100 mM NaCl. The potential physiological significance of this pathway was suggested by the demonstration that 2-BrHDA was produced by phorbol myristate acetate-stimulated eosinophils and by the demonstration that 2-BrHDA is a phagocyte chemoattractant. Taken together, the present studies demonstrate the targeting of the vinyl ether bond of plasmalogens by the reactive brominating species produced by eosinophil peroxidase and by activated eosinophils, resulting in the production of brominated fatty aldehydes. PMID- 12643285 TI - Abstracts of the 7th Interdisciplinary Slovak-Czech Toxicological Conference. Martin, Slovakia, June 24-25, 2002. PMID- 12643284 TI - Activation of myelin genes during transdifferentiation from melanoma to glial cell phenotype. AB - Induction of myelin genes occurs around birth in the last stage of Schwann cells differentiation and is reactivated in case of nerve injury. Previous studies showed that activation of the gp130 receptor system, using as ligand interleukin 6 fused to its soluble receptor (IL6RIL6), causes induction of myelin genes such as myelin basic protein (MBP) and myelin protein zero (Po) in embryonic dorsal root ganglia Schwann cells. We also reported that in murine melanoma B16/F10.9 cells, IL6RIL6 causes a shut-off of melanogenesis mediated by a down-regulation of the paired-homeodomain factor Pax3. The present work demonstrates that these IL6RIL6-treated F10.9 cells undergo transdifferentiation to a myelinating glial phenotype characterized by induction of the transcriptional activities of both Po and MBP promoters and accumulation of myelin gene products. For both Po and MBP promoters, a repression by Pax3 and stimulation by Sox10 can be demonstrated. Because after IL6RIL6-treatment, Pax3 disappears from the F10.9 cells (as it does in mature myelinating Schwann cells) whereas the level of Sox10 rather increases, we modulated the relative level of these factors and show their involvement in the induction of myelin gene expression by IL6RIL6. In addition, however, we show that a C/G-rich CACC box in the Po promoter is required for activation by IL6RIL6, as well as by ectopic Sox10, and identify a Kruppel-type zinc finger factor acting through this CACC box, which stimulates Po promoter activity. PMID- 12643286 TI - Abstracts of the 52nd Pharmacological Days. Martin, Slovakia, June 26-28, 2002. PMID- 12643287 TI - Nuclear medicine and minimally invasive surgery of parathyroid adenomas: a fair marriage. PMID- 12643289 TI - Colonoscopy turning the focus on quality. PMID- 12643288 TI - Palmitoylation of carboxypeptidase D. Implications for intracellular trafficking. AB - Covalent lipid modifications mediate protein-membrane and protein-protein interactions and are often essential for function. The purposes of this study were to examine the Cys residues of the transmembrane domain of metallocarboxypeptidase D (CPD) that could be a target for palmitoylation and to clarify the function of this modification. CPD is an integral membrane protein that cycles between the trans Golgi network and the plasma membrane. We constructed AtT-20 cells stably expressing various constructs carrying a reporter protein (albumin) fused to a transmembrane domain and the CPD cytoplasmic tail. Some of the constructs contained the three Cys residues present in the CPD transmembrane region, while other constructs contained Ala in place of the Cys. Constructs carrying Cys residues were palmitoylated, while those constructs lacking the Cys residues were not. Because palmitoylation of several proteins affects their association with cholesterol and sphingolipid-rich membrane domains or caveolae, we tested endogenous CPD and several of the reporter constructs for resistance to extraction with Triton X-100. A construct containing the Cys residues of the CPD transmembrane domain was soluble in Triton X-100 as was endogenous palmitoylated CPD, indicating that palmitoylation does not target CPD to detergent-resistant membrane rafts. Interestingly, constructs of CPD that lack palmitoylation sites have an increased half-life, a slightly more diffuse steady state localization, and a slower rate of exit from the Golgi as compared with constructs containing palmitoylation sites. Thus, the covalent attachment of palmitic acid to the Cys residues of CPD has a functional significance in the trafficking of the protein. PMID- 12643290 TI - A multi-centre North Italian prospective survey on some quality parameters in lower gastrointestinal endoscopy. AB - BACKGROUND AND AIM: To record prospectively, in a multi-centre survey, the adherence to some quality parameters of lower digestive endoscopy in everyday practice, focusing in particular on sedation and monitoring; choice of extension of endoscopic examination by the operator and relative performance in intubation of the caecum; behaviour, in some particular clinical circumstances, such as fresh haematochezia, screening and surveillance situations, functional symptoms, management of polyps. PATIENTS AND METHODS: Twenty-five centres located in Lombardia, Italy took part in the study; the duration of the survey was 2 weeks. 1,406 consecutive cases were recorded. RESULTS: The main indications to colonoscopy were fresh haematochezia (21%). abdominal pain (11.7%) and surveillance after surgery for tumours or endoscopic polyp removal (24.3%). Screening examinations accounted for 7.1% of all examinations. Conscious sedation was administered in 63.9% of patients; pulse oxymetry and cardiac frequency monitoring was used in 44.2% of cases. Total colonoscopy was planned in 84.2% of cases; the actual rate of intubation of the caecum in the cases submitted to the procedure was 84.1%. Normal results were obtained in 24.3%. In 24.3% of patients, one or more polyps were identified; 84.2% of these were removed during the first examination. Four complications (accounting for 0.28% of cases) were recorded, all after polypectomy and all successfully treated conservetively. CONCLUSIONS: The present study has shown that some attitudes in lower digestive endoscopy are sub-optimal: in particular, screening examinations are still infrequent; conscious sedation and monitoring have probably been used too rarely; intubation of the caecum was achieved in <90% of cases; the endoscopic removal of many polyps has been delayed for unexplained reasons. On the other hand, timing of surveillance after surgery for tumours or endoscopic removal of polyps has, overall, been adequate; the rate of normal examinations and the rate of complications have been satisfactorily low. PMID- 12643291 TI - Genetic dissection between silent and clinically diagnosed symptomatic forms of coeliac disease in multiplex families. AB - BACKGROUND: Coeliac disease has a large variation in clinical outcome. In addition to the classical disease with malabsorption, many individuals have a silent form, in which subject symptoms are missing but autoantibodies and mucosa lesions are identical to the symptomatic disease. AIM: To investigate whether differences in HLA DR-DQ genes explain the variation in outcome. MATERIALS AND METHODS: HLA DQ alleles were determined in 28 multiplex families with sibling pairs in which one had the symptomatic disease but the other had the silent form. RESULTS: No differences in the distribution of HLA DR-DQ haplotypes could be observed. The clinically diagnosed coeliac disease seemed to have earlier onset than silent coeliac disease. CONCLUSIONS: Results indicate that the major genetic susceptibility locus, HLA DQ, does not determine the exact clinical outcome of coeliac disease. PMID- 12643292 TI - Sorbitol H2-breath test versus anti-endomysium antibodies to assess histological recovery after gluten-free diet in coeliac disease. AB - BACKGROUND: Gluten-free diet plays a key role in treatment of coeliac disease, but it is difficult to evaluate its effect on improvement of villous architecture using sensitive non-invasive tests. AIMS: To compare sorbitol H2-Breath Test with antiendomysial antibodies in the follow-up of coeliac disease to detect histological recovery METHODS: A total of 38 consecutive patients with coeliac disease were studied. All underwent Sorbitol H2-Breath Test, antiendomysial and oesophagogastroduodenoscopy with multiple bioptic samples before diet and then 6, 12 and 18 months after gluten-free diet. Expiratory samples were collected before patients drank the test solution (5 g sorbitol in 150 ml tap water) and thereafter every 30 min for 4 hours. An increase in H2 concentration of > or = 20 ppm above fasting baseline was considered positive for sorbitol malabsorption. Antiendomysial antibodies were evaluated by the indirect immunofluorescent method. RESULTS: Antiendomysial antibodies were positive in 32/38 patients before gluten-free diet (84.21%), while they were positive in 20/34 (54.82%), 2/16 (12.5%) and 0/2 (0%) cases after 6, 12 and 18 months of gluten-free diet, respectively, no correlation being found with improvement of histological lesions (p = ns). As far as concerns sorbitol H2-Breath Test, maximal cut-off value (in ppm) decreased progressively and parallel to histological recovery during follow up. Indeed, it decreased from a mean 63 ppm before diet to 35, 19 and 12 ppm, after 6, 12 and 18 months of gluten-free diet, with a stetistical difference being found before and after (p < 0.001). Likewise, the peak value (in minutes) appeared progressively later during follow-up, parallel to histological recovery. In fact, it appeared at a mean of 119 minutes before gluten-free diet, while it appears at a mean of 164, 195 and 219 minutes after 6, 12 and 18 months on gluten free diet. A statistical difference before and after start of gluten-free diet was found also in this case (p < 0.001). CONCLUSIONS: Sorbitol H2-Breath Test is better than antiendomysial antibodies in revealing histological recovery in the follow-up of coeliac patients after the start of gluten-free diet due to its good correlation with histological damage. Moreover, it also appears to be able to detect dietary mistakes of the patients on gluten-free diet. PMID- 12643295 TI - Coeliac disease and autoimmune cholangitis: a case report. AB - Autoimmune cholangitis can be associated with other autoimmune disorders. The case is described of a 58-ear-old female who developed severe microcytic anaemia resistant to oral iron treatment. Evaluation of the patient led to the diagnosis of coeliac disease, a rarely described association. Gluten-free diet and treatment with oral haematinics led to reversal of the anaemia. PMID- 12643294 TI - Increased risk of peristomal wound infection after percutaneous endoscopic gastrostomy in patients with diabetes mellitus. AB - BACKGROUND: Results of prospective studies on the effect of prophylactic antibiotics before percutaneous endoscopic gastrostomy are conflicting. Factors for increased risk of peristomal wound infection have not been clearly identified. AIM: To evaluate the incidence of complications of percutaneous endoscopic gastrostomy and to determine the predictors of wound infection. PATIENTS AND METHODS: Percutaneous endoscopic gastrostomy was performed on 134 patients in different disease groups between January 1996 and June 2000. Medical records were carefully reviewed for demographic data, indications for percutaneous endoscopic gastrostomy, use of prophylactic antibiotics, complications and comorbid conditions predisposing to wound infection. RESULTS: Of 134 patients, 22 (16.4%) developed complications after percutaneous endoscopic gastrostomy Wound infection, the most common complication, occurred in 19 patients (14.2%) and Pseudomonas aeruginosa was the most frequently isolated microorganism. In univariate analysis, non-malignant disease and diabetes mellitus were significantly associated with peristomal wound infection after percutaneous endoscopic gastrostomy. In multivariate analysis, only diabetes mellitus was an independent risk factor for the development of peristomal wound infection after percutaneous endoscopic gastrostomy (p = 0.035) CONCLUSIONS: Patients with diabetes mellitus have a higher risk of peristomal wound infection after percutaneous endoscopic gastrostomy. PMID- 12643293 TI - Clinical comparison of high-dose interferon-alpha2b with or without ribavirin for treatment of interferon-relapsed chronic hepatitis C. AB - BACKGROUND: Interferon a with ribavirin combination therapy is effective but still unsatisfactory in the treatment of patients with interferon-relapsed chronic hepatitis C. AIMS: To compare, in a randomized, double blind, placebo controlled study, high-dose interferon-alpha2b with or without ribavirin in the treatment for interferon-relapsers. PATIENTS: A total of 52 patients with interferon-relapsed chronic hepatitis C were randomly assigned to receive 24-week treatment with interferon-alpha2b (6 MU three times per week) combined with either ribavirin (1,000 to 1,200 mg per day) or a matched placebo and then followed for an additional 24 weeks. METHODS: Hepatitis C virus RNA was detected by reverse-transcription polymerase chain reaction. For determining viral concentration, the commercial bDNA Quantiplex hepatitis C virus-RNA 2.0 assay was used. Genotyping was performed by reverse hybridization assay RESULTS: At the end of treatment, no detectable hepatitis C virus RNA levels were observed in 92% (24/26) of patients on interferon alpha2b/ribavirin and 81% (21/26) of patients on interferon alpha2b/placebo. At the end of the follow-up, a higher sustained virological response rate was seen in patients treated with interferon alpha2b/ribavirin than those treated with interferon alpha2b/placebo (69% vs 23%, p < 0.001). Patients with either initially high levels of viral concentration or with genotype 1 responded poorly. Patients who received interferon alpha2b/ribavirin treatment and in whom no hepatitis C virus RNA was detected at 4th week after treatment had 90% chance to achieve sustained virological response. CONCLUSIONS: High-dose interferon-alpha2b plus ribavirin treatment is highly effective in interferon-relapsed patients. PMID- 12643296 TI - Idiopathic dilated cardiomyopathy associated with coeliac disease: the effect of a gluten-free diet on cardiac performance. AB - An increased incidence of coeliac disease has recently been reported in patients with idiopathic dilated cardiomyopathy. This report deals with three patients with idiopathic dilated cardiomyopathy and coeliac disease who underwent clinical and laboratory evaluation to establish the effect of a gluten-free diet on cardiac performance. Two patients observed the gluten-free diet regimen very strictly, and, after a 28-month follow-up period, showed an improvement in echocardiographic parameters as well as in cardiological features and quality of life, as evaluated by the Minnesota Living with Heart Failure questionnaire and the Gastrointestinal Symptom Rating Scale questionnaire. The third patient did not observe the gluten-free diet and presented a worsening in the echocardiographic parameters and cardiological symptoms which required supplementary drug therapy. These preliminary data appear to suggest that the gluten-free diet may have a beneficial effect on cardiac performance in patients with idiopathic dilated cardiomyopathy. PMID- 12643297 TI - On-demand proton pump inhibitor therapy in patients with gastro-oesophageal reflux disease. AB - On-demand therapy is an established modality in long-term therapy with histamine 2-receptor antagonists, in cases of mild non-erosive gastro-oesophageal reflux disease. In the literature, only a few studies have specifically addressed the problem of proton pump inhibitors on-demand treatment. The evidence, so far, available suggests that this might be an effective modality of long-term treatment in the majority of patients with non-erosive gastro-oesophageal reflux disease. This treatment modality appears to be the most cost-effective and the best tolerated medical regimen for gastro-oesophageal reflux disease. It also seems to be able to restore the impairment of health-related quality of life due to gastro-oesophageal reflux disease symptoms. Although the current standard of care for patients with non-erosive gastro-oesophageal reflux disease is maintenance therapy with daily administration of a proton pump inhibitor agent, on-demand therapy, with the same drug, may be a reasonable long-term choice. The ideal proton pump inhibitors for such treatment will be those with a more rapid onset of action, more profound acid inhibition, more predictable therapeutic effect and less drug-drug interactions. Newer proton pump inhibitors, like esomeprazole, the S-chiral isomer of omeprazole, are promising drugs for on demand treatment of gastro-oesophageal reflux disease. PMID- 12643298 TI - Risk communication and the challenge of hormesis. PMID- 12643299 TI - Hormesis and risk communication. AB - Hormesis has been defined as a dose-response relationship in which there is a stimulatory response at low doses, but an inhibiting response at high doses, resulting in a U- or inverted U-shaped dose response. Toxic agents that are detrimental to human health above certain threshold levels may induce positive effects at a dose that is significantly lower than the NOAEL level. In spite of the evidence for hormesis effects, the topic is still a matter of high controversy among toxicologists. Facing this ambiguity, the article raises the question: how should risk communication be arranged in a situation of high ambiguity, lack of empirical certainty about the effects, major policy dilemmas and significant equity problems? The article summarizes the main results of risk communication research and applies these results to the question of hormesis. First, it explains the main context variables that impact on the success or failure of any risk communication program. These refer to i) the levels of the risk debate, ii) the socio-political style of regulation, iii) different types of audiences, and iv) subcultural prototypes. Secondly, the paper addresses the major functions of risk communication: i) dealing with public perception; ii) gaining trust and credibility; iii) involving stakeholders in the communication process. The last section draws some conclusions for improving risk communication on issues of hormesis. PMID- 12643300 TI - Comments on Ortwin Renn's article 'Hormesis and risk communication': considerations about uncertainity, ignorance and governance. AB - After acknowledging Renn's careful investigation and valuable insights, this paper expands on some issues that have been somewhat neglected in his account. It addresses the relationship between risk, uncertainity and ignorance in risk assessment, focusing in particular on the need to recognize that framing assumptions condition all subsequent steps. Subsequently, it discusses the implications for risk communication in the case of hormesis. The author maintains that problems of risk are strictly and irremediably intertwined with problems of governance. Therefore, she is doubtful that regulatory agencies would promote, and the public would welcome, modifications in current regimes on the basis of still limited and debated evidence of good. PMID- 12643301 TI - Commentary on hormesis and public risk communication: is there a basis for public discussions? AB - Research on radiation exposure is now focusing on microbiology and the impact of low dose exposures on cells and cell components. Eventually, this research may provide evidence to support changes in the models used to regulate human and environmental exposures. Currently, three models using older research results are subjects of interest and comparison. The linear no-threshold model, the most restrictive on behalf of public health values, dominates regulatory decision making. Alternative models (i.e., the threshold model and the hormesis model) could reduce costs of radiation management, depending upon new research results and public acceptance. Enacting a new public exposure model is a daunting task for risk communication given existing public risk perceptions and the established public decision-making processes. Each of the three prominent models must answer the question, 'what social good requires the use of this model in contrast to the others?' PMID- 12643302 TI - Hormesis and risk communication: a comment to Ortwin Renn. AB - Professor Ortwin Renn should be congratulated for authoring the definitive piece on risk communication with regard to hormesis. Most of his conclusions I agree with, specifically the importance of labelling hormesis as a possible natural effect, thereby reducing the stigmatization associated with a technical/chemical label. Rather than discussing all the points that Renn raises, in this comment I will focus on the issue of trust, a topic that Renn does examine but which I feel does not get adequate attention and which I do not completely agree with. In so doing, in my conclusions I am more optimistic than Renn is in preparing risk communication strategies regarding hormesis and other new paradigms to target audiences (defined in most instances as the general public and stakeholders). PMID- 12643303 TI - Hormesis: public health policy, organizational safety and risk communication. AB - Thirty years of research suggests low doses of toxic substances may have positive health effects. If confirmed, hormesis will imply radical changes in risk assessment and management of existing industrial toxic sources (chemical and nuclear). Renn analyses risk communication issues and positions hormesis--largely unknown to the public today--as a hypothetical risk object in society. Our comments stress the necessity to consider hormesis first as a public health issue (versus an industrial regulatory issue), to consider the impact of managerial changes upon organizational safety culture, and to assess effects on public health from the 'bad news' of toxic exposure. PMID- 12643304 TI - Chiroptical properties of acridino-18-crown-6 ligands and their complexes with chiral and achiral protonated primary (aralkyl) amine guest molecules. AB - This paper reports CD spectroscopic studies on acridino-18-crown-6 ligands (RR)-2 and 2a (see Figure 1), and their complexes with the enantiomers of alpha naphthyl)ethylamine hydrogenperchlorate (1-NEA), 1-phenylethylamine hydrogenperchlorate (PEA) and alpha-2-naphthyl)ethylamine hydrogenperchlorate (2 NEA), and also with the achiral guests (1-naphthyl)methylamine hydrogenperchlorate (1-NMA), benzylamine hydrogenperchlorate (BA), methylamine hydrogenperchlorate (MA) and 1-methylnaphthalene (1-MN). The general feature of the CD spectra of complexes of (RR)-2 with MA, BA, (R)- and (S)-PEA is the replacement of the oppositely signed 1Bb doublet of the host by one positive band near 265 nm. The CD spectra of the heterochiral and homochiral complexes of phenazino and acridino hosts (R,R)-1, 1a, (R,R)-2 and 2a with (R)- and (S)-1-NEA and 1-NMA are governed by exciton interaction. Surprisingly, the heterochial [(R,R)/(S)] complexes of the structural isomeric 2-NEA gave rise to a positive couplet in contrast to the negative couplet measured in the spectrum of the heterochiral [(R,R)/(S)] complexes of 1-NEA. PMID- 12643305 TI - Asymmetric alpha-alkylation of phenylacetates using 2-alkylamino-2'-hydroxy-1,1' binaphthyls as chiral auxiliaries. AB - alpha-Alkylation of 2-phenylacetate ester of 2-alkylamino-2'-hydroxy-1,1' binaphthyls with various alkyl iodides proceeded with good stereoselectivities. An intramolecular hydrogen bonding between N-H group and the carbonyl oxygen seemed to play an important role for asymmetric induction. The auxiliary was also applicable to Diels-Alder reaction of an acrylate. PMID- 12643306 TI - Stereochemistry of (E)- and (Z)-1,1'-dichlorobifluorenylidenes, substituted overcrowded fullerene fragments. AB - X-ray crystallographic and semiempirical PM3 and AM1 studies of 1,1'-dichlorobi 9H-fluoren-9-ylidene (5) are reported. The X-ray molecular structure of (Z)-5 indicated an approximately C2 symmetric conformation with pure twist around C9 = C9' of 40.4 degrees. The fjord regions are somewhat overcrowded: r(C8...C8') = 315.3 pm, r(Cl(1)...Cl(1') = 341.7 pm, r(C(8)...H(8')) = 259.0 pm. The four chlorine atoms of two neighboring molecules of (Z)-5 form a chain. The PM3 calculations showed that the global minimum of 5 is the C2 symmetric twisted conformation t(E)-5, which is 2.4 kJ/mol more stable than its diastereomer C2 t(Z)-5. The corresponding AM1 relative stability is reversed: C2-t(Z)-5 is 1.1 kJ/mol more stable than C2-t(E)-5. The pure twists of t(Z)-5 and t(E)-5 are 37.0 degrees and 37.2 degrees (PM3) and 40.5 degrees and 39.1 degrees (AM1). The corresponding (E) --> (Z) (PM3) and (Z) --> (E) (AM1) energy barriers of diastereomerization are 80.6 kJ/mol (PM3) and 75.8 kJ/mol (AM1). Two anti-folded local minima conformations C2-a(Z)-5 and C(i)-a(E)-5 were found to be 21.2 and 29.5 kJ/mol (PM3) and 25.8 and 35.2 kJ/mol (AM1) less stable than t(E)-5. The syn folded conformations C(S)-s(Z)-5 and C2-s(E)-5 are transition states for the enantiomerization processes of C2-tz-5 and C2-tE-5, respectively, and lay 79.8 and 94.1 kJ/mol (PM3) and 108.3 and 107.4 kJ/mol (AM1) higher in energy than their corresponding twisted conformations. An alternative pathway for enantiomerization of C2-t(E)-5 via the anti-folded achiral intermediate C(i)-a(E) has a barrier of 56.0 kJ/mol (PM3) and 68.5 (AM1). An alternative pathway for enantiomerization of C2-t(Z)-5 via C2-t(E) and C(i)-a(E) has a barrier of 80.6 (PM3) and 75.8 (AM1) kJ/mol. PMID- 12643307 TI - Determination of absolute configurations of light-atom molecules by means of direct detection of Bijvoet differences. AB - The absolute configurations of two light-atom molecules were determined sufficiently well by direct detection of Bijvoet differences. The compounds examined were (I) beta-cytidine C9H13N3O5 and (II) (S)-3-[(R)-4,4,4-Trifluoro-3 [4-methoxyphenyl]butanoyl]-4-(phenylmethyl)oxazolidin-2-one, C21H20NO4F3. Both compounds crystallize in orthorhombic system with the space group P2(1)2(1)2(1) and Z = 4. The crystal structures were carefully refined by the technique of conventional structure analysis. All possible reflections were measured on a laboratory diffractometer with Cu Kalpha radiation. The multiple-diffraction effect was often observed especially as remarkable intensity enhancement in weak reflections. After such unreliable reflections were eliminated by comparisons among the equivalent mates, data were averaged to a set of Bijvoet pairs. Afterwards, additional measurements by the psi-scan technique were tested. Since psi-scan data showed a slight systematic error probably owing to some shape effect, an artificial absorption correction DIFABS was adopted to remove the error. Small but significant intensity differences could be detected for many Bijvoet pairs, and the absolute configurations were correctly determined without ambiguity in all cases. The R and wR values for separate refinements of enantiomorphs supported the results with slight differences. Flack parameters indicated no contradictions as well. PMID- 12643308 TI - Circular dichroism and the anomeric configuration of C-glycofuranosyl-2-phenyl-2H 1,2,3-triazoles. AB - Circular dichroism spectra of a series of 4-(tetra-hydroxy-tetryl-1-yl)2-phenyl 2H-1,2,3-triazoles were measured and the sign of their Cotton effects was correlated to the absolute configuration of the hydroxyl group alpha to the triazole base moiety. Those having the D-glycero-configuration in their Fischer projection formula show positive Cotton effect and those with the L-glycero configuration show negative Cotton effect. This correlation was extended for the assignment of the anomeric configuration of the corresponding glycofuranosyl-C nucleosides. PMID- 12643309 TI - Asymmetric cyclopropanation catalyzed by a series of copper-(Schiff-base) complexes with two chiral centers. AB - A series of copper(Schiff-base) complexes with two chiral centers derived from 1,2-diphenyl-2-amino-ethanol were synthesized and applied to catalyze the asymmetric cyclopropanation of ethenes with diazoacetates. A mechanism that can explain the observed results was proposed. Some of these complexes were also efficient catalysts for asymmetric cyclopropanation of 1,1-diphenylethene with ethyl diazoacetate, affording high e.e. of up to 98.6%. An e.e. of 80.7% was achieved when no solvent was used. PMID- 12643310 TI - Cobalt complex of cinchonine: intermolecular interactions in two crystalline modifications. AB - Two crystalline modifications of cinchonine cobalt complex, C19H23Cl3CoN2O, were obtained from mixture of saturated alcohol solutions of CoCl3 x 6H2O and cinchonine. The X-ray structure analysis revealed that the asymmetric unit of one modification, CoCn1, contains only zwitterionic molecules of the complex. In the asymmetric unit of the other, CoCn2, there are two molecules of the title compound and two molecules of ethanol. The influence of the absolute configuration, the CoCl3 coordination with quinoline, and the presence of alcohol molecules on the studied structures was established by comparison of the crystal and molecular structures of both cobalt complexes with the analogous quinine complex and zinc complex of cinchonine. The interactions that dominate in the packing of the molecules in both structures are intermolecular hydrogen bonds. They form characteristic ring systems, depending on the presence of the alcohol molecules. The ring features are also related to the absolute configuration of the alkaloid. PMID- 12643311 TI - Mechanistic aspects of chiral discrimination on an amylose tris(3,5 dimethylphenyl)carbamate. AB - The separation of [2R-[2alpha(R*),3alpha]]-5-[[2-[1-[3,5-bis (trifluoromethyl)phenyl]ethoxy]-3(S)-4-fluorophenyl)4-morpholinyl]-methyl]-N,N dimethyl-1H-1,2,3-triazole-4-methanamine hydrochloride from its enantiomer was achieved on an amylose tris-3,5-dimethylphenyl carbamate stationary phase. The retention of the enantiomers is dominated by weak hydrogen bonds while the enantioselectivity is governed by other kinds of interactions, e.g., inclusion in the amylose carbamate chains. Van't Hoffplots of 1nalpha vs. reciprocal temperature were non-linear and could be divided into two linear regions. One region at low temperature (5 degrees C- approximately 20 degrees C) and another one between 25 degrees C-70 degrees C with the change in slope occurring between 16 degrees C and 20 degrees C. DSC experiments suggested that the behavior can be attributed to breakage of H-bonds triggering a conformational change. Molecular simulation indicated a correlation between the interaction energies and the elution order obtained experimentally. The most retained enantiomer (R,R,S enantiomer) interacts with the stationary phase through a hydrogen bond between the triazole proton and the C=O groups of the stationary phase, as well as through an inclusion in the cleft of the stationary phase. The other enantiomer exhibits a bifurcated H-bond between the triazolic proton and the C=O groups of the stationary phase leading to a less stable complex. PMID- 12643312 TI - The synthesis and structure of substituted dimenthyl malonate derivatives. AB - A series of substituted dimenthyl malonate derivatives were efficiently synthesized from dimenthyl malonate using a deprotonation and alkylation strategy. The elucidation of the structure of these derivatives were determined by a combination of X-ray crystallography, NMR and IR spectroscopy. PMID- 12643313 TI - Conformational analysis and TDDFT calculations of the chiroptical properties of tris [1,2-propanediolato(2-)-kappaO,kappaO'] selenium/tellurium and related compounds. AB - The tris(didentate) chelates [E(OCR1R2CR3R4O)3], with E = Se and Te, display both configurational (delta or lambda; R or S) and conformational (delta or lambda) chirality. In order to assess the contributions of these three chiral arrays to the Cotton effects of the chelates and to elucidate their stereochemistry in the gas phase and in solution, calculations of the UV and CD spectra (down to 180 nm), and also of the relative stability of the chelates, have been performed at the TDDFT/TZVP/B-P86 level. An extensive conformational analysis has supplied additional information on the relevant conformers in the conformational manifold. It was found that the dominant CD effect reflects mostly the delta/lambda twists of the three five-membered ligand rings, and less so the influence of the A/A core configuration, while the contributions of any R/S chiral carbon atoms of the ligand rings are negligible. The sign, the intensity, and the energy of this dominant CD band are found to depend on the stereochemistry of the chelates in a predictable way. Among the conformers, those with equatorially disposed methyl substituents are much preferred. These results make it possible to determine the absolute configuration (A/A) and conformation (delta/lambda) of the chelates from the CD data. PMID- 12643314 TI - Mixed cobalt(III) complexes with aromatic amino acids and diamine. Part IV. Influence of noncovalent intra-and interligand interactions on rotamer population of the S-phenylalaninato and S-tyrosinato side groups in (1,2-diaminoethane)bis (S-aminocarboxylato)cobalt(III) complexes. AB - Rotamer population of S-tyrosinato and S-phenylalaninato ligands side groups in diastereomers of (1,2-diaminoethane)bis-(S-aminocarboxylato)cobalt(III) complexes is calculated by vicinal alpha and beta proton coupling constant analysis. The effect of noncovalent intra- and interligand interactions on the population of rotamers in D20 solution is discussed. It has been established that in all the complexes investigated the most abundant is rotamer t, in which aromatic voluminous moiety and carboxylic group are in an anti position. In almost all complexes the lowest content is of rotamer g, in which these two groups are in the nearest position. Relatively high population of rotamer h in complex 5 tyr, in spite of high steric hindrances, is due to intra- and interligand NH...pi interactions. PMID- 12643316 TI - New crystalline complex of quinine and quinidine. AB - A new complex of diastereoisomeric pair, quinine and quinidine (QQd), was obtained from a mixture of saturated ethanol solutions of quinine and quinidine (0.5:1). The complex crystallises in the triclinic system, space group P1, and contains two molecules of quinine, two molecules of quinidine and four water molecules in the asymmetric unit. The X-ray structure analysis of a single crystal revealed that quinine and quinidine molecules occur in the so-called open conformation, characteristic for Cinchona alkaloids, whenever they are engaged in intermolecular hydrogen bonds. Quinine and quinidine molecules are organized in two very similar kinds of chains. In each chain the links that contain 14 membered rings can be distinguished. Within these rings quinine and quinidine molecules interact via intermolecular hydrogen bonds between the quinuclidine nitrogens and hydroxyl groups, mediated by water molecules. The links are connected with each other by hydrogen bonds between water molecules and nitrogens of the quinoline moieties, which interact via pi-pi stacking. The architecture of the hydrogen bond system in QQd, compared to those observed in the crystal structures of nonhydrated quinidine, cinchonine and cinchonidine, reveals the effect of the co-crystallizing water on the molecular packing. In nonhydrated alkaloid structures the hydrogen-bonded molecules form helical chains, different from those observed in the hydrated QQd complex and hydrated quinine toluene solvate (QTol). Comparison of QQd structure with that of QTol suggests that while the intermolecular hydrogen bonds in the system quinine-water-quinidine-water are very similar to those in quinine-water-quinine-water system, the mode of pi-pi interaction between their quinoline moieties depends on the absolute configuration of the interacting alkaloid molecules. PMID- 12643315 TI - An improved methodology for the synthesis of enantiomerically pure (S)-2,3-O cyclohexylideneglyceraldehyde. AB - An improved methodology to prepare (S)2,3-O-cyclohexylideneglyceraldehyde is described. Starting from the commercially available (L)-cyclohexylidene protected ascorbic acid the enantiomerically pure aldehyde was synthesized in only two steps in 41% overall yield. PMID- 12643317 TI - Generation of chirality by the aggregation of column structures for two-component molecular crystals composed of chloronitrobenzoic acids and p-anisidine. AB - Four two-component molecular crystals 1.2, 1.3, 1.4 and 1.5 composed of chloronitrobenzoic acids and p-anisidine as components have been prepared. Among these molecular crystals, 1.2 and 1.3 showed the generation of chirality (space group P2(1)) by aggregation of mutually parallel column structures with their 2(1) axes pointing in the same direction, and the others showed no chirality. This paper shows that the generation of chirality in a molecular crystal can be explained on the basis of the aggregation of column structures and that weaker intermolecular interactions affect the generation of chiral molecular crystal. PMID- 12643318 TI - Global anticoagulant effects of a synthetic anti-factor Xa inhibitor (DX-9065a): implications for interventional use. AB - Heparin has been conventionally used as an anticoagulant for medical and surgical indications. Because factor Xa is an essential component of the prothrombinase complex and leads to the generation of thrombin, its inhibition has become a focus of newer antithrombotic drug development. The in vitro anticoagulant profile of DX-9065a, a synthetic direct factor Xa inhibitor, was studied using activated clotting time assay, thrombelastography, and global clotting tests, such as prothrombin time (PT), activated partial thromboplastin time (aPTT), diluted aPTT, Heptest, Heptest-HI, dilute Russell's viper venom time (dRVVT), thrombin time, ecarin clotting time, and amidolytic anti-Xa assay. In addition, the effect of DX-9065a on platelet aggregation and inhibition of thrombin generation markers (FPA, F1+2, and TAT) were studied. The pharmacokinetic and pharmacodynamic profiles of DX-9065a were also studied in a non-human primate (Macaca mulatta) model. DX-9065a produced a concentration-dependent increase in the Hemochron celite ACT and HemoTec ACT. Clotting times of 538 +/- 19 and 401 +/ 12, respectively, were reached at a concentration of 25 microg/mL signifying that DX-9065a may be useful in interventional cardiological procedures. DX-9065a prolonged the r-time on thrombelastography. DX-9065a did not show any effect on adenosine diphosphate (ADP)-, collagen-, epinephrine-, and arachidonic acid induced platelet aggregation at concentrations up to 10 microgram/mL. DX-9065a exhibited a concentration-dependent prolongation of the PT, aPTT, diluted aPTT, Heptest, dRVVT, and reached the clotting times of 51.6, 132, 193, 47.9, 129.9 seconds, respectively, at a final concentration of 12.5 microgram/mL; compared to a control value of 10.6, 30.2, 41.9, 14, 32.2 seconds, respectively. DX-9065a did not affect the ecarin clotting time and thrombin time at concentrations up to 12.5 microgram/mL. Because DX-9065a prolonged the dRVVT, this may impact diagnostic screening of patients with systemic lupus erythematosus. PMID- 12643319 TI - The LONFLIT4-VENORUTON study: a randomized trial prophylaxis of flight-edema in normal subjects. AB - This study evaluated the protective effects of Venoruton on the development of flight edema in normal subjects travelling in economy in long-haul flights (8-9 hours). Edema is a relevant aspect of long-haul flights affecting both venous patients and normal subjects. Edema and flight microangiopathy were studied in a group of 164 subjects randomized (after informed consent) into two groups to evaluate prophylaxis in 8- to 9-hour flights. The treatment group received Venoruton (hydroxyethyl rutosides) 1 g twice daily for 3 days (2 days before the flight and the day of the flight). The control group received comparable placebo. Edema was evaluated with a composite edema score based on the edema tester, on the measurements of ankle circumference, volume measurements, subjective swelling, and on a discomfort score. Items 1, 4, and 5 are based on an analogue scale line (1 to 10) directly defined by the subjects before and after the flights. Of the included subjects, 151 completed the study. Dropouts were due to poor compliance, travelling, or connection problems. Age and gender distribution were comparable in the two groups as were risk factor distributions. The level of edema at inclusion was comparable in the two groups of subjects. After the flight there was an average score of 6.7 (SD 1.7) in the control group, while in the Venoruton group the score was on average 3 (SD 1.1) (p < 0.05). In the control group 77% of the subjects had an evident increase in ankle circumference and volume, which was clearly visible at inspection and associated with discomfort. In the Venoruton group, edema was clearly present only in 8% of subjects (associated with discomfort rated between 3 and 6 on the analogue scale line) and it was mild, not associated with symptoms. Therefore the control of flight edema with Venoruton was clear both considering parametric data (circumference and volume) and non-parametric (analogue scale lines) measurements. The combined evaluation of the edema score is significantly favorable for subjects under prophylaxis with Venoruton. PMID- 12643321 TI - Proposal of a flow chart for thrombosis-free oral contraceptive therapy. AB - Oral contraceptive therapy (OCT) is associated with a fourfold increase in the risk for venous thromboembolism (VTE). Women with congenital defects have a much higher risk. Because it is impossible to screen all women undergoing OCT, it is advisable to adopt some guidelines in an attempt to define the group of women who may be particularly exposed to the risk of VTE. On the basis of experience and that in the literature, two flow charts are used that are simple and reliable. The two flow charts are based on an accurate family and personal history so that an extensive investigation can be carried out only for women who show a positive family or personal history for idiopathic thrombosis. The definition of idiopathic thrombosis may be difficult, particularly in the case of family members. In case of doubt the thrombotic event should be considered as idiopathic. The two flow charts are not intended to eliminate the OCT-related VTE but represent only an attempt to decrease the prevalence of this complication. PMID- 12643322 TI - The effect of plasma depletion of platelet concentrates on platelet aggregation and phosphatidylserine expression. AB - Plasma-reduced platelet concentrates are commonly administered to prevent febrile transfusion reactions and to avoid fluid overload in neonates. Because little is known about the influence of centrifugation and resuspension on functional aspects of platelets, we examined the effects of plasma-reduction on platelet aggregation and platelet-dependent thrombin generation. Our results show that plasma reduction and resuspension of the platelet pellet in saline or plasma results in a significant reduction in platelet aggregation to a combination of the platelet agonists adenosine diphosphate and epinephrine (p < 0.001). In contrast, when a combination of the more potent agonists collagen and thrombin was used, platelet aggregation was maintained. Likewise, no decline was observed in platelet-dependent thrombin generation as measured by the functional prothrombinase assay or Annexin V binding. We conclude that centrifugation and resuspension of platelets to render the concentrate plasma-free, as a routine procedure in blood banking, variably affects in vitro platelet aggregability but does not significantly affect platelet-dependent thrombin generation. PMID- 12643320 TI - On the prophylactic and therapeutic use of danaparoid sodium (Orgaran) in patients with heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is a rare but dangerous complication of heparin prophylaxis or treatment. The present laboratory tests to measure heparin associated antibodies are not specific. The diagnosis of HIT mainly depends on the decrease in platelet count and on clinical symptoms. To evaluate clinical outcome, bleeding complications and platelet counts were evaluated in 45 patients with HIT type II (HIT II) treated prophylactically (subcutaneous injections) or therapeutically (intravenous infusion) with danaparoid. Group I included 24 patients with HIT II without thromboembolic complications who received danaparoid twice daily subcutaneously (10 IU/kg) for a mean of 16 days. Group II included 21 patients with thromboembolic complications. They were treated with intravenous danaparoid (2.6 IU/kg/h +/- 1.1) for a mean of 17 days. During subcutaneous prophylaxis, mean anti-Xa levels of 0.2 U/mL and during intravenous treatment, mean anti-Xa levels of 0.4 U/mL were reached. No deaths, amputations, or serious bleeding complications occurred, and no new thromboses were observed in both patient groups. Treatment with danaparoid led to a fast normalization of the platelet counts. This normalization occurred earlier and the concentration of platelets was higher in patients treated with intravenous doses. Danaparoid with subsequent vitamin K-antagonist treatment effectively prevents thromboembolic complications in patients with HIT. PMID- 12643323 TI - Thromboembolic disease and present oral contraception. AB - Since their introduction, oral contraceptives have been linked to an increased incidence of thromboembolic events. Epidemiologic studies have shown that women who use third-generation oral contraceptives containing desogestrel, gestodene, or norgestimate have a higher risk of venous thrombosis than women who use second generation oral contraceptives containing levonorgestrel. Women who use oral contraceptives are significantly less sensitive to activated protein C. From January 1996 to December 2001, 17,577 patients were hospitalized in our department, 177 of them (1%) had confirmed diagnosis of venous thromboembolism and 15 of those (177) were women on oral contraceptive therapy. Oral contraceptives were taken from 28 days to 18 months. No other potential congenital or acquired causes of thrombosis were present before thromboembolic events occurred in these women. The discontinuation of the pill combined with usual heparin (in one patient thrombolysis) and coumarin therapy was effective in all cases. After the discontinuation of coumarin (3-6 months), every patient was screened for hereditary thrombophilia. All women on oral contraceptive therapy had confirmed hereditary risk factors for venous thrombosis. Acquired resistance to activated protein C may explain the epidemiologic observation of increased risk of venous thrombosis in oral contraceptive users, especially in women using third-generation oral contraceptives who had other risk factors (hereditary or acquired) for venous thrombosis. PMID- 12643324 TI - Atherosclerotic and hemostatic abnormalities in patients undergoing hemodialysis. AB - Vascular events caused by atherosclerosis are the major cause of death in patients undergoing hemodialysis (HD). The relationship between the tests of atherosclerosis and hemostasis in 84 patients with HD was examined. Abnormal test results indicting the occurrence of atherosclerosis were found in 66% by the Fontaine score, in 33% by ankle blood pressures, and in 79% by aortic calcification index (ACI). When HD was prolonged, the mean Fontaine score and ACI were further increased. Particularly, the ACI tended to correlate with HD duration. The ankle-brachial index (ABI) was decreased in patients with HD duration of more than 10 years. Before HD, the plasma levels of fibrinogen, plasmin-plasmin inhibitor complex (PIC), thrombomodulin (TM), and D-dimer were increased, while the plasma levels of protein C (PC), antithrombin (AT), thrombin antithrombin complex (TAT), and tissue plasminogen activator (tPA)-plasminogen activator inhibitor-I (PAI-I) complex (tPA-PAI-1 complex) were decreased. With prolonged HD, the plasma levels of AT and PC were decreased, while those of D dimer were increased. The plasma levels of TAT and TPA-PAI-1 complex were significantly increased and those of PIC, soluble fibrin (SF) and D-dimer tended to be high in patients with less than 0.7 of ABI. The plasma levels of D-dimer, TPA-PAI-1 complex, TAT, PIC, and SF tended to be high in patients with more than 0.5 in ABI. These findings suggest that patients undergoing HD have progressive atherosclerosis and that this is associated with some hemostatic abnormalities. PMID- 12643325 TI - Protein S deficiency in repetitive superficial thrombophlebitis. AB - Protein S deficiency is one of the principal congenital thrombophilias reported. The objective of this study was to assess the prevalence of protein S deficiency in patients with repetitive superficial thrombophlebitis. Thirty-six consecutive patients, 28 female and eight male, who presented with repetitive superficial thrombophlebitis (two or more cases); age range, 17 to 58 years, mean, 38.2 years; were evaluated. Coagulometry was used to determine the biologic activity of protein S. In 5.5% of the patients with repetitive superficial thrombophlebitis, protein S deficiency was detected. PMID- 12643326 TI - Influence of the Alu-repeat I/D polymorphism in t-PA gene intron 8 on the stimulated t-PA release after venous occlusion. AB - Tissue type plasminogen activator (t-PA) is released from endothelium in both a constitutive and regulated fashion. In healthy subjects, an association between net t-PA release rate and a few t-PA gene polymorphisms, including the Alu-repeat I/D polymorphism in intron 8, was described. The possible influence of the Alu repeat polymorphism on t-PA release was evaluated after a venous occlusion test (VO) in 82 patients showing an impaired fibrinolytic capacity associated with different arterial disease or with previous venous thrombosis, and in 50 healthy controls. Euglobulin lysis time, t-PA antigen (t-PA:Ag) and activity, PAI-1 antigen and activity plasma levels were assayed before and 20 minutes after VO; the Alu-repeat I/D polymorphism was determined by PCR. Defective fibrinolysis was due to reduced t-PA release in 40 patients (t-PA group) and to PAI-1 excess in 42 patients (PAI group). No differences in both genotype distribution and allele frequencies were observed between patients and controls. The t-PA:Ag increase after VO (20/0-minute levels ratio adjusted for hematocrit) was considerably higher both in controls and in PAI group patients carrying the I allele than in the DD genotype carriers (II, ID, DD: 3.77+/-0.62, 3.43+/-0.44, 2.06+/-0.32 in controls, and 3.67+/-0.23, 2.80+/-0.50, 1.62+/-0.29 in PAI group, respectively). The difference was significant between the DD and both the ID and II genotypes in controls (p<0.05), and between the DD and II genotypes in PAI-1 group (p<0.05). A slight and nonsignificant trend of association between genotype and t-PA:Ag 20/0 ratio was seen in the t-PA group patients. In conclusion, these data suggest a possible genetic modulation of t-PA-regulated secretion. PMID- 12643327 TI - Good or poor responses of hemostatic molecular markers in patients with hematopoietic disorders after treatment of disseminated intravascular coagulation. AB - Changes of hemostatic markers in 226 patients with disseminated intravascular coagulation (DIC) and hematopoietic disorders were examined after treatment of DIC. The changes in prothrombin time (PT) ratio, fibrinogen, fibrin and fibrinogen degradation products (FDP), antithrombin, and protein C, thrombin antithrombin complex (TAT), plasmin-plasmin inhibitor complex (PPIC), and soluble fibrin monomer complex (SFMC) in all patients with DIC were significant during the clinical course of DIC, but those of D-dimer, thrombomodulin (TM), tissue factor (TF), and tissue factor pathway inhibitor (TFPI) were not. Activated partial thromboplastin time (aPTT) and PT were significantly longer in the poor response group than in good response group. Plasma levels of FDP, TAT, PPIC, SFMC, TM, and DIC score were significantly higher in poor response group than in good response. Protein C and antithrombin levels were significantly lower in poor response group than in good response group. The changes of PT ratio, fibrinogen, FDP, DIC score, antithrombin, plasmin inhibitor, and protein C were significant in the good response group, but these levels were not significant in the poor response group. The changes in plasma TAT and SFMC levels were significant in the good response group but were not in poor response group. The changes in D-dimer, TM, TF, or TFPI were not significant in both groups. These findings suggest that anticoagulant agents should be administered at levels below TAT 40 ng/mL or SFMC 300 microgram/mL in patients with DIC and hematopoietic disorders. PMID- 12643328 TI - Effects of PEG-hirudin in clotting parameters and platelet function and its interaction with aspirin in healthy volunteers. AB - The purpose of this study was to investigate the pharmacodynamics of PEG-Hirudin and its potential interactions with acetylsalicylic acid (ASA 325 mg once daily from days 1-3). In a randomized, 2-way cross-over trial, 6 healthy volunteers received PEG-Hirudin (i.v. bolus of 0.2 mg/kg + 0.02 mg/kg/h for 24 hours) and placebo (i.v. bolus + 24-hour infusion). In a further randomized, 3-way cross over trial another 9 healthy volunteers received ASA (325 mg) or oral placebo from days 1 to 3 and PEG-Hirudin (0.2 mg/kg + 0.02 mg/kg/h for 24 h) or i.v. placebo on day 3. Assessments included bleeding time (BT), collagen (1 microgram ml(-1))-induced platelet aggregation (CIPA), platelet adhesion, ecarin clotting time (ECT), activated clotting time (ACT), plasma anti-factor IIa activity (aIIa), and activated partial thromboplastin time (aPTT). Ten minutes after the PEG-Hirudin injection/starting the infusion, mean plasma concentration was 3.1 microgram/mL and aPTT, ECT, and ACT were prolonged up to 80, 309, and 233 seconds, respectively. During the last 8 hours of the 24-hour infusion mean PEG Hirudin plasma concentration was 1.3 microgram/mL. In the interaction study, ASA significantly inhibited CIPA. At 6 hours after administration, on day 3 mean BT was 6.5 minutes after PEG-Hirudin alone, 18.2 minutes after ASA alone, and 32.9 minutes after combined administration of ASA and PEG-Hirudin. PEG-Hirudin (0.2 mg/kg + 0.02 mg/kg/h for 24 hours) administered alone or together with 325 mg ASA proved to be safe in healthy volunteers. Combined use of PEG-Hirudin and ASA significantly increased the mean bleeding time compared to ASA or PEG-Hirudin monodrug administration. None of the clotting parameters or platelet function tests correlated with the prolongation of the bleeding time. PMID- 12643329 TI - Mechanical prosthetic heart valve thrombosis despite optimal anticoagulation in a patient with congenital thrombophilia (factor V Leiden). AB - Failure of anticoagulant therapy in a patient with a St. Jude heart valve in the mitral position is reported. After 8 years of optimal anticoagulation at an International Normalized Ratio between 2.5 and 3.5, a floating thrombus was discovered by chance during transthoracic echocardiography and then surgically removed. A study for congenital or acquired thrombophilia disclosed a positivity for activated protein C resistance, due to the presence of factor V Leiden. The intensity of warfarin anticoagulation was increased to provide more protection for the patient. PMID- 12643330 TI - The recent revision of the Code of Ethics for Nurses with Interpretive Statements (American Nurses Association). PMID- 12643331 TI - Illness uncertainty, perceived control, and psychological distress among adolescents with type 1 diabetes. AB - This study examined the relationship between illness uncertainty, perceived control, and psychological distress among adolescents with type 1 diabetes. Sixty eight adolescents age 13 to 18 years with type 1 diabetes completed the Children's Uncertainty in Illness Scale, the Perceived Control Scale Media Relations, and the Brief Symptom Inventory. Increased uncertainty was significantly associated with both decreased perceived control and increased psychological distress. Further analyses indicated that the relationship between illness uncertainty and psychological distress was direct and was not mediated or moderated by perceived control. These findings suggest that interventions aimed at managing uncertainty may help decrease psychological distress among adolescents with diabetes. PMID- 12643332 TI - Cancer screening promotion among medically underserved Asian American women: integration of research and practice. AB - Mammography and Pap smear tests are known to be effective early detection measures for breast and cervical cancers, respectively, but Asian Americans are reluctant to make visits for routine preventive care. Quantitative and qualitative research conducted by the Healthy Asian Americans Project (HAAP) between 1996 and 1999 indicated that Asian residents in southeastern Michigan, like the general Asian population in the US, underutilized early cancer screening programs due to cultural, psychosocial, linguistic, and economic barriers. This article reports how the HAAP's research findings guided the Michigan Breast and Cervical Cancer Control Program (BCCCP) promotion (conducted from 2000 to 2001 among medically underserved Asian women residing in southeastern Michigan), and how evaluation of the HAAP's BCCCP promotion will direct future research and health promotion programs. The article presents strategies used to improve access to cancer screening programs for diverse Asian sub-groups as well as outcomes of the 2-year HAAP's BCCCP promotion among the target population. Discussion regarding lessons and experiences gained from integration of research and practice has implications on design and implementation of the cancer screening promotion for the rapidly increasing Asian American population as well as other medically underserved minority populations in the US. PMID- 12643333 TI - The underrepresentation of Hispanics/Latinos in nursing education: a deafening silence. AB - While our national population is growing more ethnically and racially diverse, the nursing workforce and faculty remains predominantly White (non-Hispanic). Ethnic/racial minorities are seriously underrepresented in nursing and, as such, are a factor in the nursing workforce shortage. The nursing literature has addressed the need for ethnic/racial diversity for several decades, most often citing diversification as a means of providing culturally competent care. However the fact remains that the nursing population continues to be underrepresented by people of color. The diversification of the nursing profession will require culturally sensitive research and theory development regarding the recruitment and education of ethnic/racial minorities. While there is a growing body of knowledge regarding the educational needs and experiences of minorities in nursing education, much of this data generalize minorities as if they were a homogeneous group. There is a striking paucity of research regarding the underepresentation of Hispanics in nursing and the unique issues of recruitment and retention of this group in nursing education. This paper provides a review of the mostly anecdotal literature regarding the experiences of Hispanics in nursing education. It explores the underrepresentation and paucity of culturally specific knowledge regarding the recruitment, retention, and graduation of Hispanics/Latinos in nursing. The homogeneity of nursing and its consequences specifically in relation to Hispanics is discussed. The research implications are highlighted throughout. PMID- 12643334 TI - Academic talk about dementia caregiving: a critical comment on language. AB - A language that characterizes the work of informal caregivers as burdensome dominates the academic literature on dementia caregiving. Informal caregiving is a socially important activity of significant economic benefit that is both gendered female and primarily carried out by women. Words collocated with caregiving and caregivers were extracted from a sub-set of recent professional journal articles on dementia caregiving. Collocations were numerically described and grouped into three categories: (1) recipient (or what caregivers receive as a result of caregiving), (2) generative (what caregivers do or generate as a result of caregiving), and (3) demographic (the general context of caregivers and caregiving). Collocations in the recipient category were most numerous followed by demographic and generative collocations. Overall collocations suggested dementia caregiver/caregiving passivity in the context of an unrewarding, burdensome experience. Questions raised in relation to findings focus on whether the view of caregiving revealed may further inscribe oppressive practices related to disabilities, age, and gender roles. PMID- 12643335 TI - Response to "Using nursing science does not guarantee nursing excellence". PMID- 12643336 TI - Nursing information. PMID- 12643337 TI - A glimpse over the horizon. PMID- 12643338 TI - Slow response to loss of glycemic control in type 2 diabetes mellitus. AB - BACKGROUND: To achieve glycemic control in type 2 diabetes mellitus, the American Diabetes Association (ADA) recommends intensification of glucose-lowering therapy when the glycosylated hemoglobin (HbA1c) level exceeds 8.0%. OBJECTIVE: To study glycemic control before and after initiation of secondary antihyperglycemic therapy to better understand the pace and patterns of therapeutic failure and clinical responses to failure. STUDY DESIGN: A retrospective, population-based observational study. PATIENTS AND METHODS: From a 12-year-old diabetes registry of members of Kaiser Permanente Northwest, a large group-model HMO, we tracked the glycemic control histories of all 570 registrants who, in 1998, added metformin therapy to sulphonylurea monotherapy. RESULTS: The last HbA1c level before metformin use averaged 9.4%. Metabolic decompensation accelerated over time. Patients typically spent numerous months at and had several measurements of HbA1c >8.0% before a final glycemic spike to >9.0%. Persons experiencing more gradual failure accumulated greater glycemic burdens before changing therapy. CONCLUSIONS: The level of HbA1c that seemed to trigger glucose-lowering action was 9.0% or higher, not 8.0% as recommended by the ADA. A substantial hyperglycemic peak preceded change in therapy even in this relatively tightly controlled population with type 2 diabetes mellitus. Earlier therapeutic changes, but not more frequent testing, would prevent the glycemic excursions we observed. Low mean HbA1c levels in populations do not necessarily indicate that loss of glycemic control is being rapidly addressed for most patients. More research is needed to estimate the impact of these peaks on current well-being and future complications. PMID- 12643340 TI - Physician attitudes toward strategies to promote the adoption of medical evidence into clinical practice. AB - BACKGROUND: [corrected] Promoting the adoption of medical evidence into clinical practice has been advocated as one approach to improving healthcare quality and reducing medical errors. Data describing the effectiveness of different strategies to achieve this goal in real-world settings are limited. OBJECTIVE: To determine the effectiveness of selected interventions on the adoption of medical evidence into clinical practice. STUDY DESIGN: A cross-sectional survey of a random sample of physicians selected from the American Medical Association's Physician Master File. PATIENTS AND METHODS: We examined the perceived effectiveness of 7 strategies (represented by 27 individual interventions) and 5 general approaches for promoting the adoption of medical evidence into clinical practice in 1100 practicing physicians. Respondent exposure to interventions was also determined. Regression analyses were performed to identify factors that affected effectiveness ratings. Analysis of variance was used to test the hypothesis of equal mean scores across different comparison groups. RESULTS: Of 1,100 surveys mailed, 63 (5.7%) were excluded and 431 were completed (response rate = 41.6%). Mean +/- SD effectiveness scores for the 27 individual interventions on a 5-point Likert scale ranged from 2.0 +/- 0.9 (literature received from insurance companies and managed care organizations) to 4.2 +/- 0.8 (one-on-one communication with respected colleagues). Ranges for the 7 strategies were 2.6 +/- 1.0 (patient-mediated interventions) to 3.6 +/- 0.9 (educational meetings) and for the 5 general approaches were 1.98 +/- 0.9 (administrative interventions) to 3.3 +/- 0.8 (provider education). The hypothesis of equal mean effectiveness scores was rejected for all comparisons (P < .001). Frequency of exposure was the only variable to predict effectiveness (P < .001 for all regression models). CONCLUSION: From the perspective of practicing physicians, the frequency of exposure to strategies for promoting the adoption of medical evidence into clinical practice strongly affects their perceived effectiveness. PMID- 12643339 TI - How do minor changes in the definition of blood pressure control affect the reported success of hypertension treatment? AB - OBJECTIVE: In 1999, the first reporting year of the Health Plan Employer Data and Information Set, our organization noted a significant difference in the proportion of patients with controlled blood pressure (BP) when the target was changed from < 140/90 mm Hg to < or = 140/90 mm Hg. We compared these data with a second larger dataset to determine if these findings were an isolated or common phenomenon and to determine what factors influenced this difference. STUDY DESIGN AND METHODS: Subjects were drug-treated hypertensive patients. Blood pressure measurements were taken from outpatient medical records. RESULTS: The percent of hypertensive patients with controlled BP levels was significantly greater (P <.001) when the cutoff of < or = 140/90 mm Hg was used rather than <140/90 mm Hg. When a single BP measurement was used to estimate control, the percent of patients with controlled hypertension increased 12.7% simply by changing the definition to include the level of systolic BP = 140 mm Hg or diastolic BP = 90. When multiple BP readings were used to estimate control, the difference was much less pronounced (2.7%). CONCLUSIONS: When evaluating the success of hypertension treatment, defining BP control as a BP of < or = 140 mm Hg systolic and < or = 90 diastolic mm Hg will result in significantly more people having controlled BP than when <140 mm Hg systolic and <90 mm Hg diastolic are used. This difference is most likely the result of end-digit preference. The use of multiple measures reduces this effect. Policy makers and guideline authors should consider these aspects when setting BP goals. PMID- 12643341 TI - Patient reports of coverage denial: association with ratings of health plan quality and trust in physician. AB - OBJECTIVE: To evaluate whether HMO patients' reports of denial of coverage were associated with their ratings of health plan quality and trust in their physician. STUDY DESIGN: Cross-sectional survey. PATIENTS AND METHODS: Within a mixed-model HMO, we surveyed 2000 adult patients who had seen a clinician at least once during the previous year. RESULTS: Of the 2000 patients, 921 (46%) responded. Denial of coverage within the previous 12 months was reported by 64 (7%) patients. Among the denials, 42% were for specialist referral, 32% were for tests or treatments, 18% were for a certain length of hospital stay, and 8% were for a hospital admission. Patients scoring in the lowest quartile on physical functioning were more than twice as likely as other patients to report denial of coverage (12.2% vs 5.1%, P = .001). In multivariable analyses, poor physical functioning remained the only significant independent variable associated with reporting denial of coverage (odds ratio = 3.0; 95% confidence interval, 2.4, 3.6). More than half (53%) of patients reporting denial of coverage said that they had considered leaving the health plan because of concerns about quality of care. These patients also were less likely to express high trust in their primary care physician (64% vs 78%, P < .001). CONCLUSIONS: Patients with poor physical functioning were more likely to report denial of coverage. Perceived denial of coverage was associated with lower ratings of health plan quality and with significantly less trust in patients' own primary care physicians. PMID- 12643342 TI - Restoring trust to managed care, part 2: a focus on physicians. AB - Physicians have lost their trust in managed care plans because of perceived deficits in competency, concern, choice, confidentiality, and communications. To succeed in a market-driven healthcare system, managed care plans must work to restore that trust. A variety of strategies are proposed to address physician concerns related to these 5 domains of trust. Although no single strategy will restore trust, a combination can improve the relationship with these important stakeholders. PMID- 12643343 TI - Review of the newer antiepileptic drugs. AB - This review article discusses the newer anti-epileptic drugs gabapentin, lamotrigine, tiagabine, topiramate, levetiracetam, oxcarbazepine, and zonisamide. Emphasis is given to FDA-approved indications and place in therapy. The mechanisms of action and pharmacokinetics of each drug is provided and the most common adverse effects are reviewed. Clinical studies leading to FDA approval are discussed. Practical points on proper dosing and monitoring are stressed, and drug interactions are also included. PMID- 12643344 TI - New insights into the immunology and evolution of HIV. AB - Fewer than one million HIV infected individuals are currently receiving anti retroviral therapy. The limitations of such treatment have underscored the need to develop more effective strategies to control the spread and pathogenesis of HIV. Typically, naturally occurring protective immune responses provide the for such development. It is now clear however that HIV can utilise the milieu of an activated immune system to its own replicative advantage. Mobilisation of the immune response, intended to thwart the virus, may instead fuel its dissemination, 'immune escape' and spread. The immense genetic variation of HIV contributes to lack of immune control and the development of progressive disease in the majority of infected, untreated individuals. Further delineation of the intimate interactions between the HIV and the immune system will be critical and recent advances in this direction are discussed. PMID- 12643345 TI - Hypertrophic cardiomyopathy: from gene defect to clinical disease. AB - Major advances have been made over the last decade in our understanding of the molecular basis of several cardiac conditions. Hypertrophic cardiomyopathy (HCM) was the first cardiac disorder in which a genetic basis was identified and as such, has acted as a paradigm for the study of an inherited cardiac disorder. HCM can result in clinical symptoms ranging from no symptoms to severe heart failure and premature sudden death. HCM is the commonest cause of sudden death in those aged less than 35 years, including competitive athletes. At least ten genes have now been identified, defects in which cause HCM. All of these genes encode proteins which comprise the basic contractile unit of the heart, i.e. the sarcomere. While much is now known about which genes cause disease and the various clinical presentations, very little is known about how these gene defects cause disease, and what factors modify the expression of the mutant genes. Studies in both cell culture and animal models of HCM are now beginning to shed light on the signalling pathways involved in HCM, and the role of both environmental and genetic modifying factors. Understanding these mechanisms will ultimately improve our knowledge of the basic biology of heart muscle function, and will therefore provide new avenues for treating cardiovascular disease in man. PMID- 12643346 TI - Regulation of swelling-activated chloride channels in embryonic chick heart cells. AB - Swelling-activated Cl- currents, I(Cl,swell) were measured during hyposmotic shock in white Leghorn embryonic chick heart cells using the whole-cell recording of patch-clamp technique. Genistein, an inhibitor of protein tyrosine kinase (PTK), suppressed I(Cl,swell). Under isosmotic condition phorbol 12-myristate 13 acetate (PMA), an activator of PKC, elicited the Cl- current similar to that in hyposmotic solution, whereas hyposmotic shock did not elicit I(Cl,swell) in chelerythrine chloride(an inhibitor of PKC)-treated cells. Confocal microscopy experiments using FITC-phalloidin as a fluorescent label of F-actin showed that the actin network was moved from cortical region of the cell to the center after hyposmotic shock as compared with the image under isosmotic condition. When the cells were treated with cytochalasin B (CB) or cytochalasin D (CD) under isosmotic condition the disruption of the F-actin integrity was observed, and I(Cl,swell) was not elicited. With combination treatment of CB with PMA, hyposmotic solution could not elicited I(Cl,swell). The results suggested that the role of PTK, probably receptor tyrosine kinase, for regulation of I(Cl,swell) appeared to be at upstream site related to the role of F-actin. Then PKC signal pathway was activated somehow and finally change in the polymerization state of cytoskeleton led to activate the swelling-activated Cl- channels. These results demonstrate clearly that PTK, PKC and F-actin are important factors for regulation of I(Cl,swell), in embryonic chick heart cells as compared with often controversial results reported in different cell types. PMID- 12643347 TI - Ohmefentanyl stereoisomers induce changes of CREB phosphorylation in hippocampus of mice in conditioned place preference paradigm. AB - The present study was designed to determine the changes of phosphorylation of cAMP- response element binding protein (CREB) in hippocampus induced by ohmefentanyl stereoisomers (F9202 and F9204) in conditioned place preference (CPP) paradigm. The results showed that mice receiving F9202 and F9204 displayed obvious CPP. They could all significantly stimulate CREB phosphorylation and maintained for a long time without affecting total CREB protein levels. The effect of F9204 was similar to morphine which effect was more potent and longer than F9202. We also examined the effects of ketamine, a noncompetitive N-mthyl-D aspartate receptor (NR) antagonist, on morphine-, F9202- and F9204- induced CPP and phosphorylation of CREB in hippocampus. Ketamine could suppress not only the place preference but also the phosphorylation of CREB produced by morphine, F9202 and F9204. These findings suggest that alterations in the phosphorylation of CREB be relevant to opiates signaling and the development of opiates dependence. NR antagonists may interfere with opiates dependence and may have potential therapeutic implications. PMID- 12643348 TI - Construction, expression and characterization of the engineered antibody against tumor surface antigen, p185(c-erbB-2). AB - The c-erbB-2 proto-oncogene encodes a 185kDa protein p185, which belongs to epidermal growth factor receptor family. Amplification of this gene has been shown to correlate with poor clinical prognosis for certain cancer patients. The monoclonal antibody A21 which directed against p185 specifically inhibits proliferation of tumor cells overexpressing p185, hence allows it to be a candidate for targeted therapy. In order to overcome several drawbacks of murine MAb, we cloned its VH and VL genes and constructed the single-chain Fv (scFv) through a peptide linker. The recombinant scFvA21 was expressed in Escherichia coli and purified by the affinity column. Subsequently it was characterized by ELISA, Western blot, cell immunohistochemistry and FACS. All these assays showed the binding activity to extracellular domain (ECD) of p185. Based on those properties of scFvA21, we further constructed the scFv-Fc fusion molecule with a homodimer form and the recombinant product was expressed in mammalian cells. In a series of subsequent analysis this fusion protein showed identical antigen binding site and activity with the parent antibody. These anti-p185 engineered antibodies have promised to be further modified as a tumor targeting drugs, with a view of application in the diagnosis and treatment of human breast cancer. PMID- 12643349 TI - Clusterin mRNA expression in apoptotic and activated rat thymocytes. AB - Clusterin is a 75-80 kDa heterodimeric glycoprotein, that is produced in most tissues but which exact biological role is still not clear. Particularly, its role in protection or promotion of apoptosis is heavily disputed, since data supporting both views have been reported in several independent studies. To clarify this issue, and also to determine whether clusterin expression itself might be affected by apoptosis, in the present study, rat thymocytes were treated with dexamethasone, -a synthetic glucocorticoid that elicits apoptosis in thymocytes-, and clusterin mRNA expression was analyzed by semi-quantitative RT PCR before and after induction of apoptosis. Interestingly, neither the treatment with dexamethasone in vitro nor triggering of apoptosis in vivo up- regulated clusterin expression, opposing the view that clusterin is involved in apoptotic processes. On the other hand, a new clusterin mRNA isoform was detected and isolated, whose expression was restricted to freshly isolated thymocytes. This novel isoform lacks the post-translational proteolytic cleavage site and is therefore predicted to encode a monomeric protein. The biological function under normal circumstances, however, will need further investigations for clarification. While apoptosis could not modulate clusterin expression, activation of thymocytes with concanavalin A and interleukin-2 resulted in up regulation of clusterin mRNA level, indicating that clusterin expression is rather under the control of cell activation-mediated rather than apoptosis induced signals. PMID- 12643350 TI - Identification of ABA-responsive genes in rice shoots via cDNA macroar-. AB - Phytohormone abscisic acid (ABA) was critical for many plant growth and developmental processes including seed maturation, germination and response to environmental factors. With the purpose to detect the possible ABA related signal transduction pathways, we tried to isolate ABA-regulated genes through cDNA macroarray technology using ABA-treated rice seedling as materials (under treatment for 2, 4, 8 and 12 h). Of 6144 cDNA clones tested, 37 differential clones showing induction or suppression for at least one time, were isolated. Of them 30 and 7 were up- or down-regulated respectively. Sequence analyses revealed that the putative encoded proteins were involved in different possible processes, including transcription, metabolism and resistance, photosynthesis, signal transduction, and seed maturation. 6 cDNA clones were found to encode proteins with unknown functions. Regulation by ABA of 7 selected clones relating to signal transduction or metabolism was confirmed by reverse transcription PCR. In addition, some clones were further shown to be regulated by other plant growth regulators including auxin and brassinosteroid, which, however, indicated the complicated interactions of plant hormones. Possible signal transduction pathways involved in ABA were discussed. PMID- 12643352 TI - VBAC: protecting patients, defending doctors. PMID- 12643353 TI - Calf muscle therapy for Achilles tendinosis. PMID- 12643351 TI - Consequences of medical errors observed by family physicians. AB - In two studies about medical errors, family physicians reported health, time, and financial consequences in nearly 85 percent of their error reports. Health consequences occurred when the error caused pain, extended or created illness, or placed patients, their families, and others at greater risk of harm. Care consequences included delayed diagnosis and treatment (sometimes of serious health conditions such as cancer), and disruptions to care that sometimes even resulted in patients needing care in a hospital. Other important consequences were financial and time costs to patients, health care providers, and the health system generally. However, sometimes no consequence was apparent. PMID- 12643354 TI - Preventing hypotension effect of calcium channel blockers. PMID- 12643355 TI - Possible dangerous interaction of oxycontin and carisoprodol. PMID- 12643356 TI - Probable relationship between opioid abuse and heroin use. PMID- 12643357 TI - Vitamin B12 deficiency. AB - Vitamin B12 (cobalamin) deficiency is a common cause of macrocytic anemia and has been implicated in a spectrum of neuropsychiatric disorders. The role of B12 deficiency in hyperhomocysteinemia and the promotion of atherosclerosis is only now being explored. Diagnosis of vitamin B12 deficiency is typically based on measurement of serum vitamin B12 levels; however, about 50 percent of patients with subclinical disease have normal B12 levels. A more sensitive method of screening for vitamin B12 deficiency is measurement of serum methylmalonic acid and homocysteine levels, which are increased early in vitamin B12 deficiency. Use of the Schilling test for detection of pernicious anemia has been supplanted for the most part by serologic testing for parietal cell and intrinsic factor antibodies. Contrary to prevailing medical practice, studies show that supplementation with oral vitamin B12 is a safe and effective treatment for the B12 deficiency state. Even when intrinsic factor is not present to aid in the absorption of vitamin B12 (pernicious anemia) or in other diseases that affect the usual absorption sites in the terminal ileum, oral therapy remains effective. PMID- 12643358 TI - Information from your family doctor. Vitamin B12. PMID- 12643359 TI - The 'crashing asthmatic.'. AB - Asthma is a common chronic disorder, with a prevalence of 8 to 10 percent in the U.S. population. From 5 to 10 percent of patients have severe disease that does not respond to typical therapeutic interventions. To prevent life-threatening sequelae, it is important to identify patients with severe asthma who will require aggressive management of exacerbations. Objective monitoring of pulmonary status using a peak flow meter is essential in patients with persistent asthma. Patients who have a history of fragmented health care, intubation, or hospitalization for asthma and those with mental illness or psychosocial stressors are at increased risk for severe asthma. Oxygen, beta2 agonists, and systemic corticosteroids are the mainstays of acute asthma therapy. Inhaled anticholinergic medications provide additional bronchodilation. In patients who deteriorate despite usual therapeutic efforts, evidence supports individualized use of parenteral beta2 agonists, magnesium sulfate, aminophylline, leukotriene inhibitors, or positive pressure mask ventilation before intubation. PMID- 12643360 TI - Alopecia in women. AB - Alopecia can be divided into disorders in which the hair follicle is normal but the cycling of hair growth is abnormal and disorders in which the hair follicle is damaged. Androgenetic alopecia is the most common cause of hair loss in women. Other disorders include alopecia areata, telogen effluvium, cicatricial alopecia, and traumatic alopecias. The diagnosis is usually based on a thorough history and a focused physical examination. In some patients, selected laboratory tests or punch biopsy may be necessary. Topically administered minoxidil is labeled for the treatment of androgenetic alopecia in women. Corticosteroids and other agents are typically used in women with alopecia areata. Telogen effluvium is often a self-limited disorder. Because alopecia can be devastating to women, management should include an assessment for psychologic effects. PMID- 12643361 TI - Information from your family doctor. Hair loss in women. PMID- 12643362 TI - Diagnosing night sweats. AB - Night sweats are a common outpatient complaint, yet literature on the subject is scarce. Tuberculosis and lymphoma are diseases in which night sweats are a dominant symptom, but these are infrequently found to be the cause of night sweats in modern practice. While these diseases remain important diagnostic considerations in patients with night sweats, other diagnoses to consider include human immunodeficiency virus, gastroesophageal reflux disease, obstructive sleep apnea, hyperthyroidism, hypoglycemia, and several less common diseases. Antihypertensives, antipyretics, other medications, and drugs of abuse such as alcohol and heroin may cause night sweats. Serious causes of night sweats can be excluded with a thorough history, physical examination, and directed laboratory and radiographic studies. If a history and physical do not reveal a possible diagnosis, physicians should consider a purified protein derivative, complete blood count, human immunodeficiency virus test, thyroid-stimulating hormone test, erythrocyte sedimentation rate evaluation, chest radiograph, and possibly chest and abdominal computed tomographic scans and bone marrow biopsy. PMID- 12643364 TI - Childhood constipation. PMID- 12643363 TI - Delirium. AB - Delirium is characterized by an acute change in cognition and a disturbance of consciousness, usually resulting from an underlying medical condition or from medication or drug withdrawal. Delirium affects 10 to 30 percent of hospitalized patients with medical illness; more than 50 percent of persons in certain high risk populations are affected. The associated morbidity and mortality make diagnosis of this condition extremely important. Patients with delirium can present with agitation, somnolence, withdrawal, and psychosis. This variation in presentation can lead to diagnostic confusion and, in some cases, incorrect attribution of symptoms to a primary psychiatric disorder. To make the distinction, it is important to obtain the history of the onset and course of the condition from family members or caregivers. Primary care physicians must be able to recognize delirium so that the underlying etiology can be ascertained and addressed. The management of delirium involves identifying and correcting the underlying problem, and symptomatically managing any behavioral or psychiatric symptoms. Low doses of antipsychotic drugs can help to control agitation. The use of benzodiazepines should be avoided except in cases of alcohol or sedative hypnotic withdrawal. Environmental interventions, including frequent reorientation of patients by nursing staff and education of patients and families, should be employed in all cases. PMID- 12643365 TI - Photo quiz. Vesicular rash on the flank and buttock. PMID- 12643366 TI - Information from your family doctor. Memory loss. PMID- 12643367 TI - Information from your family doctor. Dementia: what are the common signs? PMID- 12643368 TI - Information from your family doctor. Grieving: facing illness, death, and other losses. PMID- 12643369 TI - CDC updates guidelines for prevention of perinatal group B streptococcal disease. PMID- 12643370 TI - Do I get to decide who should have a baby? PMID- 12643371 TI - Bayesian analysis: a new statistical paradigm for new technology. AB - Full Bayesian analysis is an alternative statistical paradigm, as opposed to traditionally used methods, usually called frequentist statistics. Bayesian analysis is controversial because it requires assuming a prior distribution, which can be arbitrarily chosen; thus there is a subjective element, which is considered to be a major weakness. However, this could also be considered a strength since it provides a formal way of incorporating prior knowledge. Since it is flexible and permits repeated looks at evolving data, Bayesian analysis is particularly well suited to the evaluation of new medical technology. Bayesian analysis can refer to a range of things: from a simple, noncontroversial formula for inverting probabilities to an alternative approach to the philosophy of science. Its advantages include: (1) providing direct probability statements- which are what most people wrongly assume they are getting from conventional statistics; (2) formally incorporating previous information in statistical inference of a data set, a natural approach which we follow in everyday reasoning; and (3) flexible, adaptive research designs allowing multiple looks at accumulating study data. Its primary disadvantage is the element of subjectivity which some think is not scientific. We discuss and compare frequentist and Bayesian approaches and provide three examples of Bayesian analysis: (1) EKG interpretation, (2) a coin-tossing experiment, and (3) assessing the thromboembolic risk of a new mechanical heart valve. PMID- 12643372 TI - Results of laparoscopic repair of giant paraesophageal hernias: 200 consecutive patients. AB - BACKGROUND: Giant paraesophageal hernias (GPEH) have traditionally required open operations. Increasingly, a laparoscopic approach is being applied to more complex esophageal surgery. Our objective was to update our growing experience with laparoscopic repair of GPEH. METHODS: We performed a retrospective review at our institution of patients undergoing laparoscopic repair of GPEH from July 1995 to July 2001. The GPEH was defined as greater than one-third of the stomach in the chest. RESULTS: Elective laparoscopic repair of a GPEH was attempted in 203 patients. Mean age was 67 years. The most common symptoms included heartburn (96 patients), dysphagia (72), epigastric pain (56), and vomiting (47 patients). Laparoscopic procedures included 69 Nissens, 112 Collis-Nissens, and 19 other procedures. There were three open conversions due to adhesions, but no intraoperative emergencies. Median length of stay was 3 days (range, 1 to 120 days). Minor or major complications occurred in 57 patients (28%). There were six postoperative esophageal leaks (3%), and 1 death. Median follow-up was 18 months. Five patients required reoperation for recurrent hiatal hernia. Excellent results were reported in 128 (84%) patients, 12 (8%) had a good result, 7 (5%) fair, and 5 (3%) poor (based on postoperative follow-up and GERD questionnaire). The mean postoperative GERD Health-related Quality of Life Score was 2.4 (scale 0 to 45; 0 = no symptoms, 45 = worst). CONCLUSIONS: Laparoscopic repair of GPEH is possible in the majority of patients with acceptable morbidity, a median length of hospital stay of 3 days and excellent intermediate-term results in an experienced center. PMID- 12643373 TI - Does continuous mucosal partial carbon dioxide pressure measurement predict leakage of intrathoracic esophagogastrostomy? AB - BACKGROUND: Gastroplasty after esophagectomy is associated with relevant morbidity due to anastomotic leakage of the esophagogastrostomy. The aim of this study was to find out whether continuous partial carbon dioxide pressure (pCO2) measurement of the gastric mucosa is an adequate method of monitoring the gastric tube during the postoperative course and of detecting patients with an anastomotic leakage. METHODS: Forty-seven patients with esophageal cancer underwent esophagectomy and gastric tube formation with intrathoracic esophagogastrostomy. Postoperatively, mucosal pCO2 of the gastric tube (pCO2i) was measured using continuous tonometry (TONOCAP, Datex Ohmeda). pCO2i was related to the arterial pCO2 (delta pCO2 = pCO2i - pCO2a). RESULTS: A total of 4,338 delta pCO2 measurements were recorded. On average, the pCO2i of each patient was monitored over a period of 92 hours. In 5 patients an anastomotic leakage of the esophagogastrostomy developed. The mean delta pCO2 of this group was 31.7 mm Hg (+/-19.3 SD) and significantly higher (p < 0.0001) than that of patients without anastomotic leakage (20.7 mm Hg +/- 12.8 SD). With a delta pCO2 cut-off point of 56 mm Hg measured for 5 hours, the sensitivity was 0.8, the specificity 0.9, and the positive predictive value 0.5. In patients with anastomotic leakage, the peak delta pCO2 preceded clinical symptoms. False positive delta pCO2 measurements (n = 4) were mainly due to severe pneumonia with long-term ventilation. CONCLUSIONS: Mucosal pCO2 measurement of the gastric tube can be used as an early indicator of a complicated postoperative course predicting anastomotic leakage of the esophagogastrostomy. PMID- 12643374 TI - Small cell carcinoma of the esophagus: the Tata Memorial Hospital experience. AB - BACKGROUND: Small cell carcinoma of the esophagus is a rare disease, characterized by aggressive progression. It has a high incidence of metastatic disease at presentation and a poor overall prognosis. Treatment protocols are not well established because of the paucity of cases and a lack of large studies. METHODS: We performed a retrospective review of all patients with small cell carcinoma of the esophagus diagnosed at the Tata Memorial Hospital between 1985 and 2001. We retrieved and analyzed data regarding demographic details, diagnosis, staging, type of treatment, and overall survival. RESULTS: Eighteen patients with a mean age of 62 years (range 48 to 80 years) diagnosed as having small cell carcinoma of the esophagus were analyzed. The group included 13 men and 5 women. Seven of the 18 patients (39%) presented with metastatic disease including 5 patients (28%) with liver metastases. Four patients were treated with surgery, with or without chemotherapy or radiotherapy. Three patients were treated with combination chemoradiotherapy, 2 patients with chemotherapy alone, and 5 patients with radiotherapy alone. Four patients with advanced disease and poor general condition were not offered any treatment. The overall median survival of our patients was 6 months. Patients treated with surgery and chemotherapy had a better overall survival. CONCLUSIONS: Small cell carcinoma of the esophagus should be regarded as a systemic disease with a high distant failure rate. Treatment strategies hence must incorporate systemic chemotherapy along with radical surgery or radiotherapy as part of a multimodality approach. PMID- 12643375 TI - Palliative management of malignant airway obstruction. AB - BACKGROUND: Obstruction of the airway due to unresectable malignant disease is a frightening condition that portends a poor prognosis. Endobronchial treatment modalities were reviewed to determine the most effective management strategy. METHODS: A 12-year retrospective review (1988 to 1999) of 121 consecutive patients with inoperable malignant airway obstruction (MAO) was performed. Sixty five patients received high-dose-rate brachytherapy (HDR) alone, 32 received HDR plus neodymium:yttrium-aluminum garnet laser (YAG) therapy, 16 received YAG only, 4 patients were stented, and 4 received photodynamic therapy (PDT). Follow-up was obtained by chart review and contact. RESULTS: Seventy-seven men and 44 women, median age 62 years (range 30 to 86 years), underwent 378 endobronchial procedures for relief of MAO. Good to excellent results were achieved in 77% (93/121) of patients. Seventy-two percent (23/32) of patients undergoing HDR plus YAG received a good to excellent result. All 8 patients receiving either stents or PDT had good to excellent palliation. There were no intraoperative deaths, but there were two in-hospital deaths. Complications occurred in 4% (5/121) of patients. Forty-four percent (53/121) of our patients were lost to follow-up. Mean survival was 6.7 months after the last treatment. CONCLUSIONS: Temporary relief of inoperable MAO can be accomplished with a number of endobronchial treatments used either singularly or in combination. The majority of patients managed with HDR, YAG, or HDR plus YAG received good to excellent short-term palliation. PMID- 12643376 TI - Endobronchial metallic stent placement for airway complications after lung transplantation: longitudinal results. AB - BACKGROUND: In lung transplant recipients, bronchial stenosis (SB) and bronchomalacia (MB) result in obstructive airway disease and allograft dysfunction due to pulmonary infection. We hypothesized that endobronchial metallic stent placement for SB and MB would result in long-term improvement in respiratory function and rates of pulmonary infection. METHODS: We studied symptomatic lung transplant recipients with bronchoscopic evidence of proximal airway complications (SB or MB) and a synchronous decline in forced expiratory volume in 1 second (FEV1) of at least 10% in the 6-month period before intervention. Stent placement was the primary intervention for SB and all focal MB lesions and for recurrent or refractory SB lesions failing a single initial attempt at balloon dilation. FEV1 and rates of pulmonary infection were assessed in the 12-month interval before and after stent placement. Spirometric evaluation was performed at 3-month intervals and compared with spirometry at the time of stent placement. The rates of pulmonary infection, determined by the number of antibiotics prescribed, was determined before and after endobronchial correction. RESULTS: Thirty recipients underwent a total of 75 procedures (50 stent insertions and 25 balloon dilations). FEV1 improved significantly after stent placement compared with base line (1.29 +/- 0.43 L) as follows: 3 months, 1.45 +/ 0.50 L, p = 0.014; 6 months, 1.59 +/- 0.57 L, p = 0.002; 12 months 1.59 +/- 0.53 L, p = 0.006. The infection rate decreased from the 12-month period preceding stent insertion to the corresponding period after stent insertion (6.97/100 days +/- 6.33 versus 5.74/100 days +/- 7.76, p = 0.018). Recurrent SB occurred in 17.3%. No life-threatening complications occurred after stent placement and no deaths were attributed to stent malfunction or malposition. CONCLUSIONS: In lung transplant recipients with SB and MB, maintenance of airway patency by stent placement is safe and resulted in improvements in lung function and reduced pulmonary infection rates for up to 1 year after their insertion. PMID- 12643378 TI - Role of thoracic surgery and interventional bronchoscopy in Wegener's granulomatosis. AB - BACKGROUND: Wegener's granulomatosis (WG) is defined as granulomatous vasculitis affecting small and medium-sized arteries and veins. Histologically, inflammatory changes with infiltration of the endothelium, fibrinoid necrosis, and formation of necrotizing granulomas are found. Pulmonary involvement is one of the cardinal features of WG and occurs in 85% of patients during the course of disease. Surgery is often required for both diagnosis and therapy. METHODS: Fifteen consecutive patients are presented to illustrate the spectrum of surgical interventions in WG. RESULTS: In 8 patients open lung biopsy, wedge resection, or segmental resection for hitherto undiagnosed infiltrate revealed WG. Eight patients presented with tracheal stenosis and all 8 underwent repetitive tracheoscopic dilation. Five patients presented with subglottic stenosis without any signs of pulmonary manifestation. All patients underwent tracheoscopic dilation, 3 in conjunction with glucocorticoid injection therapy. After multiple dilations, 1 patient still had destructing ulcerative tracheitis in which total stenting maintained airway patency. One patient received subglottic tracheal resection prior to multiple dilations; another patient, because of expiratory tracheal collapse, underwent stabilization of the membranous part of the trachea and the large bronchi with a polytetrafluoroethylene implant. CONCLUSIONS: Surgical lung biopsy in numerous patients established the final diagnosis. Thoracic surgery including bronchologic measures such as bouginage and stenting, however, also has a place in the long-term management of WG. PMID- 12643377 TI - Diagnostic thoracoscopic lung biopsy: an outpatient experience. AB - BACKGROUND: Tissue diagnosis of either interstitial lung disease or indeterminate pulmonary nodules can be obtained by either limited thoracotomy or thoracoscopic lung biopsy. Both procedures traditionally have required hospital admission. We report a series of patients undergoing outpatient thoracoscopic lung biopsy to demonstrate the safety and efficacy of this practice. METHODS: Sixty-two ambulatory patients with a clinical diagnosis of either interstitial lung disease or indeterminate pulmonary nodule(s) underwent thoracoscopic lung biopsy between June 2000 and June 2001. All procedures were performed with double-lumen endotracheal anesthesia and stapled wedge resection. Chest tubes were removed if no air leak was present and if chest radiograph demonstrated no residual pneumothorax. RESULTS: Of 62 patients undergoing thoracoscopic lung biopsy, 45 (72.5%) were discharged home within 8 hours of observation on the day of operation. Fourteen (22.5%) were discharged within 23 hours of their operation. Reasons for 23-hour observation included significant comorbidity (8), pain management (4), postoperative air leak (1), and conversion to muscle-sparing thoracotomy (1). Three (5%) required admission for prolonged air leak (2) or conversion to muscle-sparing thoracotomy (1). Diagnoses were obtained in 61 patients, including neoplasm (25), interstitial lung disease (18), granulomatous disease (7), and other (11). One patient was readmitted for pneumothorax. Patients diagnosed with nonbronchogenic pulmonary metastases were more likely to be discharged on the day of operation. No differences in age, smoking status, or preoperative pulmonary function testing were observed between patients requiring short-stay observation and those discharged immediately after operation. CONCLUSIONS: Outpatient thoracoscopic lung biopsy is safe and effective, and has become our procedure of choice for diagnosis of either interstitial or focal lung disease. PMID- 12643379 TI - Cytologically malignant margins of wedge resected stage I non-small cell lung cancer. AB - BACKGROUND: We have developed a novel test for the surgical margin of pulmonary malignant tumor using a cytologic technique (the run-across method in which a glass slide is run across the staple site), and we have assessed whether this method is useful in predicting margin relapse and prognosis. METHODS: From April 1996 to March 1999, 15 lesions of stage I non-small cell lung cancer (NSCLC) (maximum diameter ranged from 10 to 35 mm with a median of 20 mm) from 15 patients with cardiopulmonary impairment were excised without additional proximal resections. The surgical margin was examined using the run-across method. There were 8 male 7 female patients whose ages ranged from 51 to 80 years. One patient underwent video-assisted thoracic surgery and 14 underwent thoracotomy. The preoperative diagnoses of the patients were 13 adenocarcinomas, 2 squamous cell carcinomas, and 1 undiagnosed lesion (1 adenocarcinoma). The follow-up period ranged from 37 to 63 months. RESULTS: The rate of positive cytology was 47% in comparison with the rate of positive histology of 20%. There were 4 patients with margin relapse (3 of them contained negative histology margins) at a rate of 57% among the positive cytology patients in comparison with 0% among the negative cytology patients (p = 0.03). In a comparison of survival between the negative cytology group and the positive cytology group, there were no statistically significant differences. CONCLUSIONS: The run-across method is also useful in confirming complete resection. A positive cytology margin could lead to margin relapse even if a non-small cell lung cancer is resected with a negative histology margin. PMID- 12643380 TI - Pleural tent after upper lobectomy: a randomized study of efficacy and duration of effect. AB - BACKGROUND: The object of this study was to assess the efficay and maximum duration of effect of the pleural tent in reducing the incidence of air leak after upper lobectomy. METHODS: Two hundred patients who underwent upper lobectomy were prospectively randomized into two groups: 100 patients who underwent an upper lobectomy and a pleural tent procedure (group 1; tented patients) and 100 patients who underwent only an upper lobectomy and not a pleural tent procedure (group 2; untented patients). The preoperative, operative, and postoperative characteristics of both groups were compared. Then multivariate analyses were used to identify factors predictive of prolonged air leaks and their duration. The reduction of incidences of air leak in the two groups was subsequently compared during successive postoperative periods. RESULTS: No differences were detected between the two groups in terms of preoperative and operative characteristics. A significant reduction occurred in group 1 patients for the mean duration of air leak in days (2.5 vs 7.2 days; p < 0001), the number of days a chest tube was required (7.0 vs 11.2 days; p < 0.0001), the length of postoperative hospital stay in days (8.2 vs 11.6 days; p < 0.0001), and the hospital stay cost per patient (4,110 dollars vs 5,805 dollars; p < 0.0001). Logistic regression analyses showed that not having undergone a pleural tent procedure was the most significant predictive factor of the occurrence and duration of prolonged air leaks. A greater reduction in the duration of air leaks was observed before postoperative day 4 in group 1, and logistic regression analysis showed that having undergone a pleural tent procedure was the most significant predictive factor of air leaks that persisted for less than 4 days. CONCLUSIONS: Pleural tenting after upper lobectomy was a safe procedure that reduced the duration of air leaks and the hospital stay costs. The benefit from that procedure was achieved before postoperative day 4. PMID- 12643381 TI - Hernia of the lung. AB - BACKGROUND: Lung hernia is uncommon and methods of management vary. During the past 17 years, we have seen 8 patients with this condition. METHODS: Between 1984 and 2000, 8 patients with lung hernias were seen on our service. Three hernias were caused by a thoracic operation, one was due to chronic cough, and in four, the hernia was congenital, with delayed presentation. RESULTS: Three patients had minimal symptoms and were not operated on. Closure of chest wall in the other patients was accomplished by suture approximation of ribs in 4 patients and by polypropylene mesh in 1 patient. There were no recurrences, and these patients remain asymptomatic. CONCLUSIONS: Intercostal hernias are usually symptomatic and should be treated by operative closure. In supraclavicular hernias, the symptoms are usually minimal and complications are unlikely. These hernias can be left untreated, but the patients should be followed. PMID- 12643382 TI - Improving results of the modified Fontan operation in patients with heterotaxy syndrome. AB - BACKGROUND: Historically the Fontan operation in patients with single ventricle heterotaxy syndrome and atrial isomerism has been associated with high mortality. We studied whether recent modifications of the surgical technique have improved outcome. METHODS: A retrospective review of 135 patients with heterotaxy syndrome who underwent a Fontan operation between 1981 and 2000 was performed. RESULTS: There were 93 patients with right isomerism and 42 with left isomerism. Anomalies of venous return included 25 patients with extracardiac pulmonary venous connection (19%) and 37 patients with an interrupted inferior vena cava (27%). Thirty-six patients (27%) had at least moderate atrioventricular valve regurgitation. The type of Fontan procedure included 17 patients with an atriopulmonary Fontan connection, 67 with a lateral tunnel modification, 19 with an intraatrial tube graft, 25 with an extracardiac tubegraft, and 7 with an intra extra atrial tube graft. A fenestration was placed in 93 patients (78%). Early mortality was 19% before 1991, 3% since 1991, and no patient has died early since 1993. Ten-year survivals were 70% for Fontan operations before 1990 and 93% for Fontan operations after 1990. Thirty-two patients (23%) had prolonged pleural effusions. Risk factors for death included anomalous pulmonary venous connection (p = 0.02) and higher preoperative pulmonary vascular resistance (p = 0.002). Sixty-two patients (47%) had some form of early postoperative arrhythmia. At 10 years, freedom from late bradyarrhythmia and late tachyarrhythmia were 78% and 70%, respectively. Preoperative arrhythmias, older age at operation, and anatomic features were each independent predictors of late arrhythmia. CONCLUSIONS: The Fontan operation can now be performed in patients with heterotaxy syndrome with excellent survival. However, morbidity in terms of postoperative arrhythmias and prolonged pleural effusions remains significant. Fontan staging, appropriate choice of Fontan modification, aggressive treatment of concomitant malformations, and use of a baffle fenestration contribute to improved outcome. PMID- 12643383 TI - Total lymphoid irradiation for refractory rejection in pediatric heart transplantation. AB - BACKGROUND: We evaluated the role of total lymphoid irradiation (TLI) in the management of refractory rejection among pediatric heart transplant patients. METHODS: Eleven of 298 patients underwent TLI at 6 to 195 months of age and were divided into subgroups: those who survived (group A, n = 7) and those who did not survive beyond 1 year after TLI (group D, n = 4). Non-TLI recipient data were considered as the controls. RESULTS: Six out of 11 patients died eventually (54%). TLI was initiated 3 to 107 months after transplantation with a dosage of 600 to 840 cGy. The pre-TLI rejection rate (0.62 +/- 0.40 per month) was higher (p < 0.0001); however, the post-TLI rejection rate (0.24 +/- 0.65 per month) showed no significant difference from the control rejection rate. The Cox proportional hazard model found significance for TLI as a risk factor for development of posttransplant coronary artery disease (relative risk, 4.8; 95% CI, 1.1 to 21.3) and posttransplant lymphoproliferative disease (relative risk, 47.9; 95% CI, 1.6 to 1,475.3), respectively. Although the rejection rate decreased after TLI in both groups (group A pre/post, 0.51 +/- 0.31/0.06 +/- 0.08 per month; group D pre/post, 0.82 +/- 0.49/0.57 +/- 1.09 per month), significance was obtained only in group A (p = 0.018). CONCLUSIONS: TLI was an effective adjunct for reversal of refractory rejection in pediatric heart transplantation by reducing the rejection rate. Great care must be taken for the risk of development of coronary artery disease or lymphoproliferative disease. PMID- 12643384 TI - Late outcome after arterial switch operation for complete transposition of great arteries with left ventricular outflow tract obstruction. AB - BACKGROUND: Long-term follow-up of patients who underwent arterial switch operation for complete transposition of great arteries with anatomic left ventricular outflow tract obstruction (LVOTO) has rarely been brought into the focus. METHODS: Of 299 patients who underwent an arterial switch operation between January 1991 and January 2001, 23 patients had anatomic LVOTO. Age ranged from 4 days to 18 years (median 90 days) and weight ranged from 2.6 to 35 kg (median 4.3 kg). Surgical management included arterial switch operation, closure of ventricular septal defect wherever indicated, and excision of LVOTO. RESULTS: Among patients with preoperative LVOTO there were 2 early deaths and 8 patients had mild neoaortic regurgitation at the time of discharge. Follow-up ranged from 8 months to 9 years (mean 60 +/- 12 months). In 4 patients who had mild neoaortic regurgitation at discharge, the regurgitation progressed to moderate or severe degree after a follow-up of 22 to 72 months. In 1 patient mild mitral regurgitation present at the time of discharge progressed to severe mitral regurgitation. This patient subsequently underwent double valve replacement. CONCLUSIONS: Presence of preoperative anatomical LVOTO in patients undergoing arterial switch operation predicts high incidence of postoperative neoaortic regurgitation. PMID- 12643385 TI - Extracardiac conduit with a limited maze procedure for the failing Fontan with atrial tachycardias. AB - BACKGROUND: Atrial arrhythmias are a frequent late complication of Fontan procedures. Conversion to an extracardiac conduit combined with reducing right atrial size should improve hemodynamics and reduce the development of tachyarrhythmias. More effective control may be achieved by interrupting atrial arrhythmia circuits and atrial pacing. METHODS: Between May 1997 and October 2001, 6 patients underwent a revision of their intracardiac Fontan anastomosis. The conversion included an extracardiac conduit insertion, limited right atrial maze procedure, and pacemaker placement. Ages ranged from 14 to 34 years (mean, 22.8 years) at an average of 14.6 +/- 4.4 years after their original Fontan procedure. All of the patients had medically uncontrollable atrial tachyarrhythmias with markedly reduced exercise tolerance. RESULTS: All of the patients survived with an average hospital stay of 16.7 days. Exercise tolerance has improved in all 6 patients, and atrial tachycardias have either decreased (n = 2) or disappeared (n = 4). Two patients are taking antiarrhythmic medications other than digoxin. Follow-up is a mean of 28.5 months. CONCLUSIONS: Limited right atrial maze procedure, atrial size reduction, and pacemaker implantations are worthwhile additions to simple conversion to an extracardiac conduit in the failing Fontan. PMID- 12643386 TI - Cardiac troponin I in neonates undergoing the arterial switch operation. AB - BACKGROUND: Cardiac troponin I (TnI) is a sensitive and specific marker of myocardial injury, but little is known about its release after complex congenital heart surgery. We investigated whether TnI correlates with early clinical outcome in neonates undergoing the arterial switch operation (ASO) for transposition of the great arteries (TGA). METHODS: Troponin I was measured serially up to 48 hours postoperatively in 31 neonates undergoing the ASO alone (simple TGA) and 9 neonates undergoing the ASO combined with other procedures (complex TGA) (eg, closure of a ventricular septal defect) and correlated with intraoperative and postoperative clinical parameters. RESULTS: There was no mortality. Troponin I peaked at either 4 or 12 hours postoperatively in all patients (median for simple TGA = 3.4 ng/mL, interquartile range 2.4 to 4.6; median for complex TGA = 4.7 ng/mL, interquartile range 3.2 to 6.8, p = 0.20). Peak TnI correlated with the durations of inotropic support (r = 0.54, p < 0.001), ventilation (r = 0.51, p < 0.01), and intensive care unit stay (r = 0.50, p < 0.01). The duration of cardiopulmonary bypass, aortic cross-clamping, and circulatory arrest did not correlate with the peak or total TnI release. The duration of aortic cross clamping correlated poorly with the duration of inotropic support (r = 0.40, p < 0.05). The complex TGA group had longer aortic cross-clamp times, required more postoperative inotropic support, and had significantly higher total TnI release compared with the simple TGA group. CONCLUSIONS: There are weak but statistically significant correlations between peak TnI and clinical outcome. Complexity of the defect and ischemic times may be as useful to predict outcome in this group of patients. PMID- 12643387 TI - Hemodynamics and early clinical performance of the St. Jude Medical Regent mechanical aortic valve. AB - BACKGROUND: The St. Jude Medical Regent valve is the next-generation bileaflet aortic prosthesis, modified from the currently marketed St. Jude Medical mechanical valve to achieve a larger geometric orifice without changing the existing design of the pivot mechanism or blood-contact surface areas. The present study reports the hemodynamic and early clinical results of an ongoing multicenter trial investigating the performance of the Regent valve. METHODS: Between July 1998 and July 2001, 361 patients at 17 centers in North America and Europe underwent implantation of a Regent mechanical aortic valve prosthesis. Clinical status was prospectively recorded, and echocardiography with Doppler was performed at discharge and at 2 months, 6 months, 1 year, and 2 years after operation. RESULTS: Follow-up to date is 300 patient-years (average, 0.8 +/- 0.7 years per patient; range, 0.0 to 2.7 years). There were low rates of clinical adverse events. Mean gradient at 6 months was 9.7 +/- 5.3 mm Hg, 7.6 +/- 5.2 mm Hg, 6.3 +/- 3.7 mm Hg, 5.8 +/- 3.4 mm Hg, and 4.0 +/- 2.6 mm Hg, respectively, for 19-mm, 21-mm, 23-mm, 25-mm, and 27-mm valves; effective orifice area was 1.6 +/- 0.4 cm2, 2.0 +/- 0.7 cm2, 2.2 +/- 0.9 cm2, 2.5 +/- 0.9 cm2, and 3.6 +/- 1.3 cm2, respectively. Indexed effective orifice area was equal to or greater than 1.0 cm2/m2 for all valve sizes. Left ventricular mass index decreased significantly between early postoperative (165.9 +/- 57.1 g/m2) and 6-month follow-up (137.9 +/- 41.0 g/m2; delta = -28.0 +/- 49.1 g/m2; p < 0.0001). CONCLUSIONS: The St. Jude Medical Regent aortic valve has excellent hemodynamics and early clinical results, with rapid and significant left ventricular mass regression. Long-term clinical assessment is ongoing. PMID- 12643388 TI - Ten-year echocardiographic and clinical follow-up of aortic Carpentier-Edwards pericardial and supraannular prosthesis: a case-match study. AB - BACKGROUND: There are little comparative data on Carpentier-Edwards supraannular and pericardial second-generation bioprostheses. The aim of this work was to compare their hemodynamic and clinical outcomes in patients with aortic stenosis. METHODS: We conducted a retrospective study including 150 patients operated on for aortic stenosis between 1989 and 1993. Patients undergoing aortic valve replacement with either a Carpentier-Edwards supraannular or pericardial prosthesis were matched for sex (49% male), age (72 +/- 8 years), body surface area, valve size, associated procedures, and left ventricular ejection fraction. RESULTS: Mean follow-up was 6.5 +/- 3.3 years, giving a total follow-up of 983 patient-years. Thirty-day mortality and 10-year actuarial survival were, respectively, 8% and 51% in the supraannular group and 6.7% and 43.4% in the pericardial group. At 10 years, freedom from thromboembolism, structural failure, and all valve-related events were, respectively, 88.7%, 88.9%, and 68.7% in the supraannular group and 85%, 100%, and 82.2% in the pericardial group. There were four (5.3%) structural failures, and four (5.3%) reoperations for degeneration (n = 3) and endocarditis (n = 1) in the supraannular group. Freedom from structural dysfunction or reoperation was 87.3% in the supraannular group and 100% (p < 0.05) in the pericardial group. Echocardiographic review of 62 of 76 survivors (81.5%) demonstrated a trend toward a better hemodynamic profile of pericardial valves at the end of follow-up. CONCLUSIONS: Ten years after aortic valve replacement for aortic stenosis, Carpentier-Edwards pericardial prostheses give comparable and probably better results than Carpentier-Edwards supraannular prostheses. PMID- 12643389 TI - Aortic valve disease with severe ventricular dysfunction: stentless valve for better recovery. AB - BACKGROUND: Stentless bioprostheses and homografts show better hemodynamic profiles compared with conventional stented bioprostheses and mechanical valves. Few data are available on stentless aortic valve implantation for patients with severe left ventricular dysfunction. The aim of this retrospective study was to assess the potential benefits of stentless aortic valve implantation for patients undergoing isolated aortic valve replacement with left ventricular ejection fraction < or = 35%. METHODS: From November 1988 through March 2000, 53 patients (45 men and 8 women, aged 64.2 +/- 15.2 years) with a LVEF < or = 35% (mean EF, 28.7 +/- 5.4%) underwent isolated, primary aortic valve replacement for chronic aortic valve disease. Twenty patients received stentless aortic valves and 33 patients received conventional stented bioprostheses and mechanical valves. Predictive factors for LVEF recovery at echocardiographic follow-up (36.2 +/- 32.1 months) were analyzed by simple and multiple regression analysis. RESULTS: There were no significant differences between groups in early and late mortality. Stentless aortic valve implantation required a longer aortic cross-clamp time (p = 0.037). The stentless aortic valve group showed a better LVEF recovery (p = 0.016). Stentless aortic valves had a larger indexed effective orifice area compared with conventional stented bioprostheses and mechanical valves (p < 0.0001). A smaller indexed effective orifice area (p = 0.0008), chronic obstructive pulmonary disease (p = 0.015), and implantation of a conventional stented bioprosthesis or mechanical valve (p = 0.016) were related to reduced LVEF recovery by univariate analysis. A larger indexed effective orifice area (p = 0.024) was an independent predictive factor for a better LVEF recovery by multivariate analysis. CONCLUSIONS: Stentless aortic valve implantation for patients with severe left ventricular dysfunction, even if technically more demanding, is a safe procedure that warrants a larger indexed effective orifice area leading to an enhanced LVEF recovery. PMID- 12643390 TI - Nineteen-millimeter prosthetic aortic valves allow normalization of left ventricular mass in elderly women. AB - BACKGROUND: Implantation of small aortic valve prostheses has been reported to be associated with impaired left ventricular (LV) mass regression and incomplete resolution of symptoms although these data have been generated largely with male patients. Therefore we sought to determine the clinical and hemodynamic outcomes of female patients who received a 19-mm aortic valve. METHODS: Between May 1995 and December 2000, 38 female patients (average age 73 years, range 42 to 89) underwent isolated aortic valve replacement (AVR; n = 22) or AVR plus coronary artery bypass graft surgery (CABG; n = 16) with a 19-mm aortic prosthesis. The average New York Heart Association (NYHA) class was 3.08 and of the 26 patients who had angina, 47.2% were in CCS class III or IV. Clinical and echocardiographic follow-up was done an average of 33.4 months (8 to 72) after surgery. RESULTS: Operative mortality was 10.5%. Overall survival at an average of 33 months was 71.1%. The average NYHA class was 1.52 +/- 0.34 postoperatively (p < 0.001 versus preoperative) and 95% had no anginal symptoms or were in Canadian Cardiovascular Society class I. The LV mass index showed significant regression (114 +/- 11 g/m2 to 89 +/- 9 g/m2, p = 0.001) despite an effective orifice area index (EOAI) of 0.64 +/- 0.09 cm2/m2. CONCLUSIONS: Despite a very small EOAI, elderly female patients with 19-mm prosthetic aortic valves can experience a satisfactory improvement in symptoms and normalization of LV mass. This finding suggests that small prosthetic aortic valves continue to have an application in contemporary cardiac surgical practice. The current perception of patient-prosthesis mismatch may need to be reconsidered for select populations. PMID- 12643391 TI - Results of valve-sparing aortic root reconstruction in 158 consecutive patients. AB - BACKGROUND: This study assesses the durability and clinical outcome of valve sparing aortic root reconstruction using the reimplantation technique in a single center cohort. METHODS: From July 1993 to July 2001, 158 patients underwent replacement of the ascending aorta with native valve reimplantation. Mean age of patients was 52 +/- 17 years (9 to 84 years), 103 were men (65%). Thirty-four patients (22%) suffered from Marfan's syndrome. Aortic dissection Stanford type A was present in 29 patients (19%) (22 acute, 7 chronic), and concomitant partial or total arch replacement was necessary in 57 patients (36%). One or more additional procedures were performed in 28 patients (18%). Mean follow-up was 36 +/- 25 months (0.4 to 96 months). RESULTS: Thirty-day mortality was 3.8% (6 patients), but only 2.2% in elective patients. Mean bypass time was 169 +/- 50 minutes (99 to 440 minutes), aortic cross-clamp time was 129 +/- 31 minutes (79 to 205 minutes). In patients undergoing arch replacement, circulatory arrest was 26 +/- 18 minutes (7 to 99 minutes). During follow-up, there were 5 (3.3%) cardiac-related late deaths. Grade of aortic insufficiency (AI) decreased from 2.3 +/- 1.1 (0 to 4) preoperatively to 0.23 +/- 0.44 (0 to 2) postoperatively (p < 0.0001). Six patients required aortic valve replacement, 4 of those due to progressive AI. Average grade of AI increased significantly to 0.42 +/- 0.61 (0 to 3) at latest evaluation (p = 0.002). Two patients experienced a transient ischemic attack within the first postoperative week. No further thromboembolic complications were noticed. All patients presented with a favorable exercise tolerance. CONCLUSIONS: The aortic valve reimplantation technique achieves excellent clinical outcome with few complications even in complex pathologies. Lack of anticoagulation and favorable durability encourage wider and earlier use of this technique. PMID- 12643392 TI - Perioperative risk factors for mortality in patients with acute type A aortic dissection. AB - BACKGROUND: This study was undertaken to identify the perioperative risk factors for death in patients with acute type A aortic dissection (AADA). METHODS: Between 1993 and 2001, 108 consecutive patients (86 men; mean age, 53 years) underwent emergent operations for AADA. All patients but 2 underwent replacement of the ascending aorta with an open distal anastomosis during a period of hypothermic circulatory arrest. In addition, 22 patients had hemiarch and 5 had total arch replacement. Aortic root was replaced in 20 and repaired with gelatin resorcinol-formaldehyde glue in 39 patients; aortic valve was separately replaced in 3, resuspended in 24, and remained untouched in 22 patients. RESULTS: Overall in-hospital mortality was 25%. Mortality rate was significantly higher in patients with preoperative dissection complications than in those without (21/36 [58%] vs 6/72 [8%], p < 0.001). In multivariate analysis, predictors of mortality were presence of rupture, renal failure, and intestinal malperfusion, duration of cardiopulmonary bypass > or = 200 minutes, blood loss > or = 500 mL, and transfusion of blood > or = 4 units. Location of the intimal tear, extent of the replacement, type of the aortic root repair, and duration of hypothermic circulatory arrest did not emerge as predictors of mortality. CONCLUSIONS: Major determinants of surgical mortality in patients with AADA are preoperative complications. Earlier diagnosis remains essential to improve the survival rate. PMID- 12643393 TI - Multimodal protocol influence on stroke and neurocognitive deficit prevention after ascending/arch aortic operations. AB - BACKGROUND: Various techniques are used for brain protection during aortic surgery. Rather than evaluate each factor separately, we evaluated the early outcome of a multimodal protocol (mannitol, thiopental, MgSO4, lidocaine, CO2 field flooding, Leukoguard filter, head ice packing, electroencephalographic arrest at 20 degrees C, alpha-stat, increasing right subclavian artery cannulation, and antegrade/retrograde brain perfusion) for brain protection. METHODS: Prospectively collected data were analyzed on 403 ascending or arch aortic operations including 199 (49%) arch replacements conducted between July 25, 1991, and September 25, 2001. The mean age was 61.6 years (range 22 to 91 years); 48 (12%) had Marfan syndrome; 141 (35%) had dissection; 134 (33%) had composite grafts inserted; and 138 (34%) had concurrent coronary bypasses performed. RESULTS: Stroke occurred in 2.0% (8/403) (3 permanent, 5 transient), clinical neurocognitive deficits in 2.5% (10/403) either by testing or patient complaint 2 to 3 weeks after surgery, and 98% (395/403) were 30-day survivors. Univariate predictors of stroke, neurocognitive decline, or death were the following: for stroke, aorta symptom severity grade (1 to 4) (p = 0.001), pump time (p = 0.001), arrest time (p = 0.001), macroscopic atheroma (p = 0.041), concurrent descending/thoracoabdominal aneurysm (p = 0.036), and highest blood rewarming temperature (p = 0.043); for neurocognitive decline, degree of cooling (p = 0.046), pump time (p = 0.001), cooling time (p = 0.001), day extubated (p = 0.042), and antegrade brain perfusion (p = 0.004); for death, pump time (p = 0.001) and clamp time (p = 0.011). The multivariable independent predictors of stroke, neurocognitive decline, or death were the following: for stroke, aorta symptoms grade (p = 0.025), peripheral vascular disease (p = 0.043), and pump time (p = 0.015); neurocognitive decline, preoperative New York Heart Association dyspnea class (p = 0.022), pump time (p = 0.05), arrest time (p = 0.06), day extubated (p = 0.042), and antegrade perfusion (p = 0.023); and for death, pump time (p = 0.018). CONCLUSIONS: Pump time continues to be the most important predictor of adverse events. The benefit of antegrade or retrograde perfusion remains unproven, partly because of the low event rate (< 2.5%) but may be beneficial for prolonged circulatory arrest. Embolic material either from macroscopic atheroma, descending or thoracoabdominal aneurysms, or associated with peripheral vascular disease, increases the risk of stroke. Preoperative symptoms influence outcome. PMID- 12643394 TI - Aneurysm formation after patch aortoplasty repair (Vossschulte): reoperation in adults with and without hypothermic circulatory arrest. AB - BACKGROUND: Aortic aneurysm formation is common after patch aortoplasty repair of coarctation of the aorta. Its incidence varies between 5% and 38%. The majority of patients show progressive aneurysmal dilation within 6 to 18 years and reoperation is necessary to avoid rupture of the aneurysm. METHODS: Ten patients were reoperated on for patch aneurysm formation. Femorofemoral cardiopulmonary bypass (CPB) with a heparinized system was used in all patients. Decision to initiate hypothermic circulatory arrest (HCA) was made intraoperatively. All patients received a Dacron graft replacement of the aneurysmatic thoracic aorta. RESULTS: HCA was initiated in 5 patients owing to extreme adhesions in vicinity to the aneurysm. There was no significant intergroup difference regarding time interval after first operation, age, operation time, and postoperative blood loss. Only minor neurologic events were present in 2 patients with cross-clamping the aorta. CONCLUSIONS: Patch aneurysms after Vossschulte aortoplasty can safely be operated on with femorofemoral CPB. Initiation of HCA is recommended to prevent rupture of the aneurysm during preparation and injury of adjacent nerves and vessels. PMID- 12643395 TI - Do left ventricular assist device (LVAD) bridge-to-transplantation outcomes predict the results of permanent LVAD implantation? AB - BACKGROUND: Implantable left ventricular assist devices (LVADs) were designed for permanent implant, but we began their use for bridge-to-transplant (BTTx) to study their safety and effectiveness. We review our experience in order to compare the BTTx lessons learned with the outcomes and goals of permanent implants. METHODS: From December 1991 until January 2002, 264 patients received 277 LVADs for BTTx. We analyzed temporal trends in pre-LVAD patient factors and device-specific time-related complications. RESULTS: Survival to transplant was 69%. Adverse event analysis demonstrated a high risk of infections (0.56, 1.28, and 1.88 per patient at 30 days and 3 and 6 months). HeartMate devices were more prone to infection than Novacor devices (p < 0.0001). Cerebral infarctions occurred less commonly than infections (0.15, 0.25, 0.30 at 30 days and 3 and 6 months), were more common in Novacor than HeartMate (p = 0.0001), and were decreased by the new Novacor Vascutek conduit (p = 0.07), but these were still slightly higher than the HeartMate (p = 0.04). Device failures occurred in 21 instances (all but one were in HeartMate devices [p = 0.04 vs Novacor]), but have significantly decreased (p < 0.0001) in HeartMate since 1998. CONCLUSIONS: Infections and device durability limit the chronic use of the HeartMate device, but device failures are decreasing. Novacor has fewer problems with infection and durability, and the new Vascutek conduit will reduce, but not eliminate, strokes. PMID- 12643396 TI - Intracoronary E-/L-selectin blockade reduces neutrophil infiltration in heart transplantation. AB - BACKGROUND: This study examined the effect of local intracoronary delivery of a unique monoclonal antibody (mAb) to both E- and L-selectin (EL-246) on neutrophil infiltration after global ischemia during cardiac transplantation. METHODS: In 12 ovine heart transplants, allograft coronary arteries were locally perfused with EL-246 (n = 6), or isotype-matched control antibodies (n = 2) or saline (n = 4). At 24 hours posttransplant, myocardium was analyzed for neutrophil infiltration and myocardial water content. RESULTS: The mean number of intramyocardial neutrophils per area (PMN/hpf) was greatly reduced in the allografts perfused with EL-246 (3.45 +/- 0.4 PMN/hpf), compared with an average 6.5 +/- 0.97 PMN/hpf in control hearts (p = 0.004). Peripheral leukocyte counts were unaffected; myocardial water content was not significantly reduced. CONCLUSIONS: Local perfusion of cardiac allografts with blocking antibody EL-246 before reperfusion significantly reduced the neutrophilic infiltration that occurs early after transplantation. Prohibiting neutrophil-endothelial adhesion and transmigration may be useful in decreasing neutrophil-dependent post-reperfusion injury in transplantation and routine cardiac surgery. PMID- 12643397 TI - Intraoperative high-rate atrial pacing test as a predictor of atrial fibrillation after coronary artery bypass surgery. AB - BACKGROUND: Atrial fibrillation (AF) is a common complication after coronary artery bypass grafting (CABG) operations. The aim of this prospective trial was to test the hypothesis that intraoperative high-rate atrial pacing may induce AF by mimicking rapid atrial tachycardia and can identify the patients at risk for postoperative AF. METHODS; Eighty patients having on-pump CABG without additional procedures were included in the study. After cannulation but before initiation of cardiopulmonary bypass two pacing wires were placed on the lateral surface of the right atrium. The right atrium was paced with the rate of 200 beats per minute for 10 seconds. If the patient was in sinus rhythm after the high-rate pacing, the pacing test was repeated with the rate of 250 and finally 300 beats per minute. RESULTS: Postoperatively AF developed in 28 patients (35%). The high-rate atrial pacing test induced AF in 27 patients (33.7%). Of the 28 patients who experienced AF during the postoperative period, 17 patients were inducible in the atrial-pacing test (sensitivity 0.61). Of the 52 patients who did not develop AF postoperatively, 42 patients were not inducible in the atrial-pacing test (specificity of the test was 0.81). Positive and negative predictive values of the test were 0.63 and 0.79, respectively. CONCLUSIONS: The intraoperative high rate atrial pacing test turned out to be a simple, safe, and fast way to identify the patients at risk for AF after CABG. The diagnostic accuracy of this test is sufficient to identify a group of patients to whom prophylactic treatment could be directed. PMID- 12643398 TI - Holter changes resulting from right-sided and bilateral infrastellate upper thoracic sympathectomy. AB - BACKGROUND: We tested the hypothesis that no right-sided dominance exists after infrastellate surgical upper-thoracic sympathectomy. We aimed to confirm whether a significant bradycardia was constant and only dependent on the right side. METHODS: We performed 24-hour Holter electrocardiographic recordings in 12 patients referred for bilateral sympathectomy. Surgery was performed at two distinct times allowing for the study of the consequences of unilateral right and bilateral sympathectomy. RESULTS: Heart rate was 77 +/- 8 beats per minute before surgery on the 24-hour recording and significantly decreased after bilateral (67.8 +/- 6.5 beats per minute; p < 0.05) but not after unilateral right sympathectomy. Consistently spectral analysis variables significantly changed after bilateral surgery but showed no right-sided dominance. Little effect of sympathectomy was found on the QT interval, which tended to decrease after bilateral sympathectomy. CONCLUSIONS: Patients should be informed of the bradycardia resulting from sympathectomy. No right-sided dominance can be found consistently with the random distribution of substellate cardiac fibers reported in anatomic studies. PMID- 12643399 TI - Revascularization in severe ventricular dysfunction (15% < OR = LVEF < OR = 30%): a comparison of bypass grafting and percutaneous intervention. AB - BACKGROUND: We sought to determine the optimal approach to revascularization of patients with severe left ventricular (LV) dysfunction. METHODS: We conducted a single-center observational study of 117 consecutive patients who had severe LV dysfunction (15% < OR = LV ejection fraction < OR = 30%) and underwent either coronary artery bypass grafting (CABG, n = 69) or percutaneous revascularization (n = 48) between 1992 and 1997. RESULTS: The CABG group was younger (62 versus 67 years, p = 0.026), and fewer previous bypasses (7% versus 40%, p < 0.0001) and fewer prior percutaneous revascularizations (16% versus 42%, p = 0.0019) were noted. More vessels were revascularized (3 +/- 0.8 versus 1.5 +/- 0.7, p < 0.0001), and revascularization was more complete by CABG (84% versus 48%, p < 0.0001). Morbidity and mortality at 30 days were similar, and there was no significant difference in 3-year survival (73% versus 67%), although 3-year cardiac event-free survival (52% versus 25%, p = 0.0011) and 3-year target vessel revascularization-free survival (71% versus 41%, p < 0.0001) were significantly better in the CABG group, and LV ejection fraction was significantly improved after CABG. In the subgroup of patients 65 years of age or older and those without proximal left anterior descending coronary artery lesions, significant benefit of CABG in cardiac event-free and target vessel revascularization-free survival disappeared. CONCLUSIONS: We found that in clinically selected patients with severe ventricular dysfunction, CABG compared with percutaneous revascularization achieves more complete revascularization, improved LV function, fewer cardiac events, and fewer target vessel revascularizations, but does not affect mid-term survival. A prospective controlled trial with defined criteria for treatment assignment is warranted to confirm our results regarding the two revascularization strategies in patients with severe LV dysfunction. PMID- 12643400 TI - Effect of off-pump coronary surgery with right ventricular assist device on organ function and inflammatory response: a randomized controlled trial. AB - BACKGROUND: Right ventricular assist devices (RVADs) have been proposed to improve exposure of the coronary arteries in off-pump surgery. In this study we investigated the impact of the A-Med RVAD on inflammatory response and organ function in patients undergoing coronary artery bypass grafting. METHODS: Sixty patients were prospectively randomized to conventional surgery with cardiopulmonary bypass (CPB) and cardioplegic arrest, beating heart surgery (off pump), or beating heart surgery with the RVAD. Serial blood samples were collected postoperatively, for analysis of inflammatory markers, troponin I, protein S100, and free hemoglobin. Renal tubular function was assessed by measuring urine N-acetyl-glucosaminidase activity. RESULTS: No hospital deaths or major postoperative complications occurred in the study population. Interleukin 6, interleukin-8, C3a, and troponin I levels after surgery were significantly higher in the CPB group compared with the off-pump and RVAD groups. Free hemoglobin levels immediately after the operation, peak and total S100 levels, and N-acetyl-glucosaminidase activity were also significantly higher in the CPB group. CONCLUSIONS: Off-pump coronary revascularization, with or without RVAD, reduces inflammatory response, myocardial, neurologic, and renal injury, and decreases hemolysis when compared with conventional surgery with CPB and cardioplegic arrest. PMID- 12643401 TI - Assessment of ascending aorta using epiaortic ultrasonography during off-pump coronary artery bypass grafting. AB - BACKGROUND: Use of an aortic partial clamp for proximal anastomosis during off pump coronary artery bypass is known to increase the risk of fatal complications. The purpose of this study was to assess the management of the ascending aorta evaluated with epiaortic ultrasonography during off-pump coronary artery bypass. METHODS: Intraoperative ultrasonography of the ascending aorta with a 10-MHz probe was performed consecutively in 155 patients undergoing off-pump coronary artery bypass between August 1999 and July 2001. The findings from ultrasonography, surgical modifications, and operative results were analyzed. RESULTS: In 54 patients (34.8%), epiaortic ultrasonography showed atherosclerotic findings in the anterior side of the ascending aorta (group A). The remaining 101 patients had either normal findings or atherosclerotic findings in only the posterior side (group NA). A proximal anastomosis to the aorta was preoperatively planned in 117 patients (group A, 42; group NA, 75). In group A, a graft modification without clamping was implemented in 29 patients (24.8% of 117 patients), whereas the clamp site was modified to a different segment in 13 patients (11.1% of 117 patients). In all 75 patients in group NA, partial clamping was used in the standard fashion. There were no cerebral infarctions or operative deaths related to partial clamping. However, aortic dissection occurred in 1 patient in group NA. CONCLUSIONS: In 35% of patients undergoing off-pump coronary artery bypass, epiaortic ultrasonography identified atherosclerotic findings in the anterior wall of the ascending aorta. This study suggests that revascularization without aortic manipulation during off-pump coronary artery bypass is indicated in as many as 25% of patients. PMID- 12643402 TI - Assessment of an aortosaphenous vein graft anastomotic device in coronary surgery: clinical experience and early angiographic results. AB - BACKGROUND: Until now technologic evolution in coronary bypass surgery has focused on extracorporeal circulation, on operation without extracorporeal circulation, and on the exposure of the operative site. Recently a one-shot anastomotic device for the proximal anastomosis in coronary surgery was developed. We investigated whether the use of the aortic connector system (ACS) could facilitate the creation of aortosaphenous vein graft anastomoses in myocardial revascularization. METHODS: From November 2000, 40 ACS devices were used in 36 consecutive patients (mean age 70.7 +/- 8.9 years); 12 patients (33.3%) underwent surgery on pump and 24 patients (66.6%) off pump; 50 distal anastomoses were performed. In all cases the connection with the ascending aorta was created before the distal anastomoses because of the necessity to slide the saphenous vein graft (SVG) over the vein transfer sheath. Intraoperative graft function was tested measuring blood flow by Doppler analysis. Postoperative evaluation of the anastomotic patency was carried out by early angiography in 34 patients (94.7%) but was excluded in 5 patients (5.3%) with extensive extracardiac vascular occlusive disease. RESULTS: Of 38 AC (95%) evaluated, 36 (94.7%) functioned properly. The end-to-side proximal anastomosis without aortic clamping is instantaneous, the quality of anastomoses was highly rated, no additional stitches were required, and all coronary arteries could be reached. Intraoperative quantity flow was measured by Doppler analysis and all but one showed good flow. Early postoperative angiography demonstrated good patency of the grafts in all cases but 2 (5.3%). At 1-year follow-up, 1 patient died of stroke; all other patients remained free of symptoms and no reoperation was required. CONCLUSIONS: The use of ACS makes end-to-side anastomosis rapid, effective, and reproducible while eliminating aortic cross clamping; it opens a new era in beating or nonbeating coronary surgery. Long-term results are mandatory to confirm our favorable preliminary results. PMID- 12643403 TI - Analysis of mortality within the first six months after coronary reoperation. AB - BACKGROUND: Identify risk factors associated with mortality following repeat coronary revascularization (redoCABG) within the first 6 months following surgery. METHODS: Five hundred forty-one redoCABG patients (1987 to 1998) were studied by univariant and multivariant analysis. Mortality was assessed at three different points: hospital mortality (A) (36/541, 6.7%); mortality at 6 months (C) (75/541, 13.9%); and outpatient perioperative mortality, which is a death occurring from the time of hospital discharge to 6 months postoperatively (B) (39/541, 7.2%). RESULTS: Diabetes, hypertension, peripheral vascular disease, renal insufficiency, lung disease, myocardial infarction (MI) before the first operation, MI between the first and redoCABG, lack of sinus rhythm, no IMA graft, acute/emergency operation, perfusion time, and perioperative MI were all identified as risk factors related to early mortality. MI before the first operation, antegrade cardioplegia, and the time period 1987 to 1992 all influenced hospital mortality (A). Diabetes, hypertension, renal insufficiency, lung disease, and valvular heart disease all influenced the outpatient mortality up to 6 months. Independent predictive factors for early mortality were: age more than 69 years; diabetes; vascular insufficiency; chronic lung disease; MI between first and redoCABG; no IMA-graft; acute preoperative MI; emergency operation; perfusion time; perioperative MI; and the time period 1987 to 1992. Risk factors for in-hospital death included MI between the first and redoCABG, cardiopulmonary bypass time, and the time period 1987 to 1992. Diabetes is an important risk factor during the outpatient perioperative phase. Emergency surgery and perioperative MI predict mortality regardless of the time period (A, B, or C). CONCLUSIONS: Early mortality after redoCABG is influenced by many variables during the first 6 months following surgery. Understanding these factors and their time course may better help to assess the true risk associated with reoperation for recurrent coronary insufficiency. PMID- 12643404 TI - Outcomes experience with off-pump coronary artery bypass surgery in women. AB - BACKGROUND: It has been well documented that women have higher morbidity and mortality rates than men following coronary artery bypass graft (CABG) surgery. In view of this evidence, we investigated the following question: compared with on-pump CABG surgery, is there benefit to off-pump CABG surgery in women? METHODS: Our investigation analyzes patient mortality and 13 procedure complications controlling for 35 variables representing patient characteristics and comorbid conditions, and for procedure characteristics for a population of 16,871 consecutive women undergoing off-pump and on-pump CABG surgery at 78 hospitals for the period January 1998 to June 2001. RESULTS: Mean comparisons reveal that the mortality rate for women undergoing off-pump CABG surgery is nearly a percentage point lower than for women undergoing on-pump surgery (3.12 vs 3.90; p = 0.052). The complication rates for all complications analyzed (shock/hemorrhage, neurologic, cardiac, respiratory, renal, acute renal failure, adult respiratory distress syndrome, implant infection, postoperative infection, septicemia, pneumonia, and peripheral vascular) were lower for women off-pump than women on-pump with the exception of mechanical complications. Logistic regression results reveal, after controlling for 35 relevant patient characteristics, comorbid conditions and procedure characteristics, that women undergoing on-pump CABG surgery experience a 42% higher mortality rate (p = 0.0239) than women undergoing off-pump CABG surgery. CONCLUSIONS: Evidence suggests that off-pump CABG surgery may be better for women than on-pump CABG surgery because it appears to reduce mortality and respiratory complications, shorten lengths-of-stay, and increases discharges directly home. None of the 12 other complications investigated demonstrated an advantage for women undergoing on-pump surgery relative to those receiving off-pump surgery. PMID- 12643405 TI - Excess coronary artery bypass graft mortality among women with hypothyroidism. AB - BACKGROUND: The impact of thyroid disease on patients undergoing coronary artery bypass grafting has been reported in only small series of selected patients. METHODS: We investigated 30-day mortality of patients on thyroxine replacement therapy undergoing isolated coronary artery bypass grafting from 1993 to 2000 and identified variables of importance for outcome. RESULTS: A total of 3,631 patients (606 women) had isolated coronary artery bypass grafting of whom 58 patients (30 women) were treated for hypothyroidism. The mortality rate was higher among women with thyroxine replacement (16.7%, 95% confidence interval [CI] 5.6 to 34.7) than those without thyroxine replacement (5.9%, 95% CI 4.1 to 8.2; p = 0.02) and no difference between men with (3.6%, 95% CI 0.1 to 17.8) and without (2.6%, 95% CI 2.0 to 3.2) thyroxine treatment (p = 0.8). Intake of diuretics (p < 0.001) was directly associated with mortality whereas intake of aspirin (p = 0.01), levothyroxine dose (p = 0.03), and serum thyroxine level (p = 0.01) were inversely associated with mortality among women on thyroxine replacement. CONCLUSIONS: Women on thyroxine replacement therapy undergoing coronary artery bypass grafting had an increased mortality rate. We speculate that insufficient thyroid hormone replacement could partly play a role in this outcome. PMID- 12643406 TI - Comparative study of functional responses and morphometric state of distal radial arteries in male and female. AB - BACKGROUND: Differences can exist in terms of physiology and morphology of blood vessels on the basis of gender. Radial artery is now considered to be the second choice for coronary artery bypass grafting. However, there is a lack of comparative studies on the function and morphometery of radial arteries in female and male patients. METHODS: Radial arteries from 9 female and 9 male patients undergoing coronary artery bypass grafting were used to compare the effects of vasoconstrictors, noradrenaline and 5-hydroxytryptamine, as well as the influence of endothelium-dependent (with methacholine) and endothelium-independent (with sodium nitroprusside) relaxations. Furthermore, morphomteric measurements of smooth muscle thickness, lumen perimeter, lumen area, and intima area (including plaque) of distal radial arteries from female and male patients were also made. RESULTS: Radial arteries from female patients when compared to male patients were significantly more sensitive to the actions of noradrenaline, and somewhat more sensitive towards the actions of 5-hydroxytryptamine. However, no significant differences were found between the relaxant effects of methacholine in radial arteries of female and male patients. In contrast, radial arteries from female patients when compared to male patients were significantly less sensitive to the relaxant effects of sodium nitroprusside. Morphometric measurements of blood vessels from female and male patients revealed that vessels obtained from female patients had a smaller lumen area and perimeter than vessels from male patients. In contrast, there were no significant differences between tunica intima area (including plaque area) or smooth muscle thickness in radial arteries of female patients when compared to male patients. However, the radial arteries from female patients had a significantly greater ratio of tunica intima area (including plaque) to lumen area when compared with radial arteries from male patients. CONCLUSIONS: Differences exist between the functional behavior and morphometery of radial arteries of female and male patients. It is possible that postbypass, radial artery graft may show different characteristics in female versus male patients. PMID- 12643407 TI - Does vacuum-assisted venous drainage increase gaseous microemboli during cardiopulmonary bypass? AB - BACKGROUND: Vacuum-assisted venous drainage enables adequate drainage through small-diameter cannulas but concerns are that it results in more gaseous microemboli delivered to the patient. METHODS: Five identical embolus detectors monitored the propagation of entrained air through a cardiopulmonary bypass (CPB) model. The ability of the CPB circuit to remove gaseous microemboli was studied with vacuum-assisted venous drainage and gravity siphon venous drainage using different pump speeds and rates of gaseous microemboli delivery. RESULTS: Under all conditions entrained venous air resulted in the detection of gaseous microemboli in the perfusate after the arterial filter. In blood-primed circuits, increased flow rates and higher levels of vacuum-assisted venous drainage were independently associated with increased gaseous microemboli counts in the arterial line. Vacuum-assisted venous drainage at -40 mm Hg did not significantly increase gaseous microemboli activity when compared with gravity siphon venous drainage at 4 L/min flow rate. CONCLUSIONS: Vacuum-assisted venous drainage at 40 mm Hg does not statistically reduce the ability of the CPB circuit to remove gaseous microemboli at lower pump rates. High levels of vacuum and increased pump flow rates should be avoided. Air should not be introduced into the venous line. PMID- 12643408 TI - Diazoxide amelioration of myocardial injury and mitochondrial damage during cardiac surgery. AB - BACKGROUND: Recently, we have shown that the selective opening of mitochondrial ATP-sensitive potassium channels with diazoxide significantly decreases myocardial injury. The purpose of this study was to determine the effects of diazoxide on apoptosis and the mechanisms modulating apoptosis and myocardial injury in a blood-perfused model of acute myocardial infarction. METHODS: Pigs (32 to 42 kg) undergoing total cardiopulmonary bypass underwent left anterior descending coronary artery occlusion for 30 minutes. The aorta was cross-clamped and magnesium-supplemented potassium cold-blood cardioplegia (DSA; n = 6) or magnesium-supplemented potassium cardioplegia containing 50 micromol/L diazoxide (DZX; n = 6) was administered, followed by 30 minutes of global ischemia and 120 minutes of reperfusion. Left ventricular tissue samples from DSA and DZX hearts were obtained after reperfusion. Apoptosis was determined by TUNEL, caspase-3 and PARP cleavage, and caspase-3 activity. Bax and bcl-2 levels were determined and tissue morphology was examined by light and transmission electron microscopy. RESULTS: Apoptosis, as estimated by TUNEL-positive nuclei/3,000 myocardial cells, was 120.3 +/- 48.8 in DSA hearts and was significantly decreased to 21.4 +/- 5.3 in DZX hearts (p < 0.05 vs control). Caspase-3 and poly-ADP-ribose polymerase cleavage and pro-apoptotic bax protein levels were significantly decreased with diazoxide (p < 0.05 vs DSA). Light and transmission electron microscopy indicated severe disruption of tissue with capillary dilatation, mitochondrial cristae damage, and evidence of increased presence of mitochondrial granules in DSA as compared with DZX hearts. CONCLUSIONS: The addition of diazoxide (50 micromol/L) to cardioplegia significantly decreases regional myocardial apoptosis and mitochondrial damage, and provides an additional modality for achieving myocardial protection. PMID- 12643409 TI - Preconditioning: myocardial function and energetics during coronary hypoperfusion and reperfusion. AB - BACKGROUND: Ischemic preconditioning (IP) is gaining more acceptance as a protective method in beating heart surgery. Yet it remains controversial whether preconditioning can attenuate myocardial dysfunction during reperfusion after severe coronary hypoperfusion. We examined this issue and also the issue of whether this protection is mediated by adenosine A1 receptors. METHODS: In isolated, blood-perfused rabbit hearts, the effects of IP (3 minutes of no flow ischemia and 8 minutes of reperfusion) during 30 minutes of coronary hypoperfusion and 60 minutes of reperfusion were investigated. In two groups (n = 8 each) with and without (control group) preconditioning, ventricular function was assessed by load-insensitive measures: slope of the end-systolic pressure volume relation (Emax), slope of the stroke work/end-diastolic volume relation (Mw), and end-diastolic pressure-volume relation. External efficiency was calculated, and contractile efficiency was assessed using the reciprocal of the myocardial oxygen consumption-pressure-volume area relationship. To investigate the possible role of adenosine, the adenosine A1 receptor antagonist DPCPX (2.5 micromol/L) was administered before preconditioning in a third group (n = 7). RESULTS: The effects of hypoperfusion on systolic function, diastolic function (dP/dtmin, end-diastolic pressure-volume relation), external efficiency, and contractile efficiency were similar in both the IP and control groups. Lactate efflux was significantly reduced after preconditioning (p = 0.02). During reperfusion, recovery of systolic function and coronary flow were significantly improved in the IP group compared with controls: aortic flow, 85% versus 63% (p = 0.01); dP/dtmax, 91% versus 67% (p = 0.001); pressure-volume area, 97% versus 68% (p = 0.01); Emax, 74% versus 62% (p = 0.03); and Mw, 94% versus 84% (p = 0.04). Release of creatine kinase was reduced in the IP group, 9.6 +/- 1.3 U x 5 min(-1) x 100 g(-1) wet weight, versus controls, 12.7 +/- 2.7 U x 5 min(-1) x 100 g(-1) wet weight (p = 0.04). During reperfusion, contractile efficiency (p = 0.03) and external efficiency (p = 0.02) recovered better in preconditioned than in untreated hearts. Recovery was less pronounced in the DPCPX group compared with the IP group (p, not significant). CONCLUSIONS: The results, derived from load insensitive measures, confirm that IP provides protection after episodes of severe hypoperfusion by attenuating systolic dysfunction without improving diastolic dysfunction and reduces the severity of anaerobic metabolism as well as ischemic injury. Contractile efficiency and external efficiency both indicate improved energetics after IP (oxygen utilization by the contractile apparatus). The protective effect, at least in part, is mediated by adenosine A1 receptors. PMID- 12643410 TI - Warm and tepid cardioplegia: do they provide equal myocardial protection? AB - BACKGROUND: Cardiac troponin I (CTnI) has been shown to be a marker of myocardial injury. The aim of this prospective, randomized study was to compare intermittent antegrade warm cardioplegia with tepid blood cardioplegia in patients undergoing first elective coronary artery bypass graft, using CTnI release as the criterion for evaluating the adequacy of myocardial protection. METHODS: Seventy patients were randomly assigned to one of two cardioplegia groups. CTnI concentrations were measured in serial venous blood samples drawn immediately before cardiopulmonary bypass and after aortic unclamping at 6, 9, 12, and 24 hours. Analysis of covariance with repeated measures was performed to test the effect of the type of cardioplegia and time on CTnI concentration. RESULTS: The total amount of CTnI released (8.23 +/- 20.5 microg in the warm group and 3.19 +/- 2.4 microg in the tepid group) was not statistically different (p = 0.23). The CTnI concentration did not differ for any sample in either of the two groups when adjusted on ejection fraction and the number of preoperative myocardial infarctions (p = 0.06). No patient in the tepid group versus 4 patients in the warm group showed CTnI evidence of perioperative myocardial infarction (p = 0.12). CONCLUSIONS: Our study showed no preference for warm or tepid cardioplegia in terms of myocardial protection, either for clinical or biological data. PMID- 12643411 TI - The use of routine chest X-ray films after chest tube removal in postoperative cardiac patients. AB - BACKGROUND: The use of routine postoperative chest x-ray films (CXRs) for postoperative cardiac patients has been challenged, suggesting that only clinically indicated CXRs be obtained. The removal of chest tubes has been used as an indication for CXRs. Our hypothesis is that routine postoperative chest tube removal CXRs are not indicated in the asymptomatic postoperative cardiac patient. METHODS: Charts of 1,021 consecutive postoperative median sternotomy patients were reviewed, focusing on postoperative findings of CXRs, clinical evaluations, and interventions. Those who died prior to tube removal were excluded from the study. RESULTS: Tubes were removed on postoperative days 1 to 7 (average, 1.45 days). The two groups of patients were comparable in age, gender, procedure, and co-morbidity (p > .01). Seven hundred three patients underwent routine postoperative tube removal CXRs. Abnormal findings were present in 282 patients. Resultant therapeutic intervention was undertaken in 13 patients and 9 were symptomatic. No imaging after routine postoperative CXRs was conducted in 283 patients. These patients remained asymptomatic and required no intervention. Fourteen patients had clinically indicated CXRs after chest tube removal. Two of these patients had additional tubes placed, and 1 patient had follow-up films. In total, there was a 1.5% incidence of therapeutic intervention after chest tube removal. All patients were discharged without further sequelae of their tubes. CONCLUSIONS: Omission of routine postoperative chest tube removal CXRs in postoperative cardiac patients is safe. The removal of chest tubes in these patients is not an indication for CXRs. PMID- 12643412 TI - Atricure bipolar radiofrequency clamp for intraoperative ablation of atrial fibrillation. AB - PURPOSE: This work reports our initial evaluation and clinical experience with the Atricure bipolar radiofrequency (RF) system for intraoperative ablation of atrial fibrillation (AF). DESCRIPTION: The Atricure system includes a bipolar RF clamp and an RF generator. Lines of conduction block are created by delivery of RF energy (75 volts, 750 milliamps) to tissue compressed between the jaws of the clamp. Energy delivery is continued until the lesion is transmural, indicated by steady and reduced conductance between electrodes in the clamp. EVALUATION: The Atricure device was used for intraoperative treatment of AF in 120 patients. Lesion sets varied, as the device was used for pulmonary vein isolation, creation of right atrial lesions, or both. There were no device-related complications. Initial incomplete pulmonary vein isolation, identified by pacing, was uncommon, occurring with very thick atrial tissue and when tightening the clamp caused tissue to become folded upon itself in the jaws or to extrude beyond the jaws of the clamp. In such cases, reapplication of the RF clamp produced complete pulmonary vein isolation. CONCLUSIONS: The Atricure bipolar RF system facilitates surgical treatment of AF. Proper application of the clamp is essential to ensure continuous transmural lesions. PMID- 12643413 TI - Congenital unilateral pulmonary artery agenesis and aspergilloma. AB - Isolated unilateral pulmonary artery agenesis is a rare condition, which in most patients is asymptomatic. Occasionally patients present with symptoms that are nonspecific and not necessarily attributable to disease of the respiratory system. In these individuals the clue to the diagnosis is found in a plain chest roentgenogram, often revealing a hyperlucent contracted hemithorax. We present an unusual case of isolated unilateral pulmonary artery agenesis associated with the opportunistic organism Mycobacterium kansasii and Aspergillus fumigatus in which the diagnosis was made 10 years after initial presentation. Clinicians should be aware of this condition and include it in their differential diagnosis of a hyperlucent lung field on the chest roentgenogram. PMID- 12643414 TI - Pericardial hemorrhage caused by costal exostosis. AB - This report describes a 21-year-old man who presented with pericardial hemorrhage due to a puncture wound of the pericardium by an inward-facing exostosis of the left fifth rib. PMID- 12643415 TI - Accessory spleniculi in the right hemithorax. AB - Accessory spleniculi are present in the thoracic cavity without a history of trauma due to anomalies in the development of spleen. We report the case of a 62 year-old woman with hereditary spherocytosis and previous splenectomy with an incidental mass on a chest radiograph and an indeterminate diagnosis on needle biopsy. The probable sequence of embryological events that may explain the anatomic presence of splenic tissue in the thorax is discussed. PMID- 12643416 TI - Endobronchial closure of postoperative bronchopleural fistula using vascular occluding coils and n-butyl-2-cyanoacrylate. AB - We report herein 2 patients with intractable postoperative bronchopleural fistula with empyema after lobectomy or subsegmentectomy. The patients underwent several treatments including thoracotomy, but the fistula closure was not successful. Finally, the bronchopleural fistula was successfully treated by endobronchial closure using vascular occluding coils and n-butyl-2-cyanoacrylate (Histoacryl). PMID- 12643417 TI - Diclofenac (Voltaren)-induced pneumonitis after chest operation. AB - We report a rare case of nonsteroidal anti-inflammatory drug-induced pneumonitis in a 72-year-old man taking diclofenac for wound pain after pulmonary resection. The pneumonitis and pleural effusion were predominant on the operative side and resolved rapidly after the diclofenac was discontinued. The diagnosis of drug induced pneumonitis was based on a lymphocyte stimulation test that was positive for diclofenac sodium and negative for other drugs. This case report demonstrated that surgeons should be aware of the possibility of pneumonitis induced by a nonsteroidal anti-inflammatory drug. PMID- 12643418 TI - Anticoagulation with bivalirudin during cardiopulmonary bypass in cardiac surgery. AB - Heparin is the standard agent used for systemic anticoagulation during cardiopulmonary bypass in cardiac operations. Alternatives are needed when patients with heparin-induced thrombocytopenia type II are encountered. We present a patient with a clinical picture of heparin-induced thrombocytopenia type II who was effectively anticoagulated with bivalirudin, a direct thrombin inhibitor, during cardiopulmonary bypass for a cardiac operation. PMID- 12643419 TI - Systemic-to-pulmonary artery shunt thrombosis in a neonate with factor V Leiden mutation. AB - We present the case of a newborn male with D-transposition of the great arteries, ventricular septal defect, and subpulmonary stenosis who experienced two episodes of systemic-to-pulmonary artery shunt thrombosis. Hematologic evaluation revealed heterozygous factor V Leiden mutation. This report emphasizes the importance of evaluation for inherited coagulation disorders in pediatric patients with unexpected thrombotic complications. PMID- 12643420 TI - Activated recombinant factor VII for control of diffuse bleeding after implantation of ventricular assist device. AB - Patients with prolonged severe cardiogenic shock requiring implantation of a biventricular assist device may develop diffuse bleeding due to alteration of hepatic and renal function and subsequent coagulopathy. Bleeding control in these patients may be difficult despite massive use of blood products. We report on the successful use of recombinant activated factor VII for control of massive bleeding after implantation of a biventricular assist device in a patient with prolonged severe cardiogenic shock. PMID- 12643421 TI - Endovascular management of a ruptured mycotic aneurysm of the innominate artery. AB - Mycotic aneurysms of the innominate artery are infrequent lesions and, as such, represent challenging surgical problems. We describe herein a case of a ruptured mycotic innominate artery aneurysm, which developed after radical neck dissection and radiation therapy for tonsillar carcinoma. The aneurysm was successfully excluded from the systemic circulation with endoluminal placement of a covered stent, with efficacy confirmed by vascular imaging at 6 months follow-up. The patient suffered no permanent neurologic sequelae. Long-term follow-up and chronic antibiotic therapy will be necessary to avoid infection of the covered stent in this high-risk surgical patient. PMID- 12643422 TI - Open surgical intervention to recurrent aortic dissection after endovascular stent grafting. AB - We report the case of a 49-year-old man who received open-heart surgery for recurrent aortic dissection after endovascular stent grafting. Stent grafting had been successfully performed in the acute phase. Recurrent dissection became obvious 5 months later, and at the same time, aneurysmal change was detected between the left subclavian artery and the proximal end of the stent graft. We employed a "Y arch" surgical procedure and "elephant trunk" technique to treat, and the entry tear was completely sealed and the aneurysm was excluded by elephant trunk segment. We believe that this approach could be a new option for treatment for complicated aortic aneurysms. PMID- 12643423 TI - Treatment of a fistula at the distal anastomosis after Bentall operation with endoluminal covered stent. AB - A 25-year-old Marfan patient was operated on for an acute type A aortic dissection that was complicated twice by false aneurysms at the distal suture line. At the third episode a covered endoprosthesis was inserted in the ascending aorta between the coronary ostia and the inominate artery. The postoperative course was uneventful and a control computed tomographic scan showed complete occlusion of the false aneurysm. This attractive technique should be considered versus an open-heart operation in selected patients. PMID- 12643424 TI - Median sternotomy and extended left anterior thoracotomy for repair of traumatic aortic transection with ruptured right atrium. AB - A 22-year-old man presented with traumatic aortic transtion associated with rupture of the right atrium and underwent urgent median sternotomy to repair the right atrium. A T-shaped extended left anterior thoracotomy was performed, and ruptured descending thoracic aorta was repaired under total bypass. A Y-shaped connector was inserted in the arterial catheter to allow cannulation of both ascending aorta and femoral arteries. A 4-cm long Hemoshield graft was used to repair the aortic transection. The patient made a full recovery and was discharged 13 days after the accident. PMID- 12643425 TI - Off-pump cardiac revascularization after a complex stab wound. AB - Penetrating cardiac injuries are among the most dramatic and lethal of all injuries. High survival rates in selected reports have given the erroneous impression that the lethality of cardiac injuries has diminished in recent years. In a review of 1,198 cases of penetrating cardiac trauma from South Africa, only 6% of patients arrived at hospital alive. Many patients with proximal coronary artery injuries present without signs of life. Cardiopulmonary bypass is advocated as a means of salvage, but mortality rates are high. We present a case of arterial revascularization using off-pump technology. PMID- 12643426 TI - Use of intraaortic balloon pump in left ventricle rupture after mitral valve replacement. AB - Over 6 years of a single surgeon experience, 3 patients had left ventricle rupture following mitral valve replacement, despite preserving the posterior leaflet. The valve was re-replaced on bypass in all patients. Intraaortic balloon pump was inserted electively before coming off bypass. There were no intraoperative deaths, reexploration, or excessive bleeding. An intraaortic balloon pump is an ideal adjuvant to left ventricle repair for ruptured ventricle following mitral valve replacement on cardiopulmonary bypass. PMID- 12643427 TI - Extracardiac total cavopulmonary connection using a Y-shaped graft. AB - We report on 17-year-old Fontan candidate with a severely distorted central pulmonary artery (PA) who underwent a successful extracardiac total cavopulmonary connection using a Y-shaped bifurcated graft. A nonanatomic pathway from the inferior vena cava to the left PA was constructed and positioned anterior to the ascending aorta. The other arm was used as a conduit between the inferior vena cava and the right PA. All procedures were performed under temporary venous bypass without cardiopulmonary bypass. PMID- 12643428 TI - Sinus of Valsalva aneurysm. PMID- 12643429 TI - Cardiac hydatid cyst located in the interventricular septum. PMID- 12643430 TI - Tracheal reconstruction with autogenous composite nasal septal graft. AB - We describe lateral resection of tracheal tumors and repair of the defect with a composite graft of nasal septal mucosa and cartilage. The preliminary experience with this technique in 3 patients was satisfactory. The procedure can be used for patients who are not candidates for standard resection and end-to-end anastomosis. PMID- 12643432 TI - Double-breasted repair of the posterior mitral valve leaflet. AB - The double-breasted repair of the posterior leaflet of the mitral valve is an alternative technique for correction of mitral regurgitation in selected patients. This new technique has the advantage of avoiding distortion of the posterior annulus and simplifies the repair, especially in complex posterior leaflet prolapse. PMID- 12643431 TI - Polytetrafluoroethylene monocusp valve technique for right ventricular outflow tract reconstruction. AB - Nonvalve transannular patch repair of right ventricular outflow tract obstructive (RVOTO) defects results in pulmonary insufficiency, which can contribute to early postoperative right ventricular dysfunction. In both animal and clinical studies, monocusp valves provide perioperative RVOT competence and improved right ventricular functional characteristics. In these reports, monocusp leaflet construction with 0.1-mm polytetrafluoroethylene (PTFE) appeared equal, or superior, to biologic monocusp valves. Construction of the polytetrafluoroethylene monocusp valve is an inexpensive and straightforward way to create a competent RVOT in a variety of RVOTO anomalies. Based on our clinical experience, it effectively prevents short-term and significantly reduces midterm pulmonary insufficiency without evidence of stenosis. PMID- 12643433 TI - Free right internal thoracic artery composite graft: an option in left anterior descending artery grafting? AB - Myocardial revascularization using bilateral internal thoracic arteries (ITA) decreases the risk of reinterventions and provides potential survival benefit. From May 1996 to April 2000, 1,057 patients underwent myocardial revascularization using skeletonized bilateral ITAs. A free right ITA as a composite graft was used for the left anterior descending artery grafting in 38 (3.6%) cases when the left ITA was not long enough to reach the left anterior descending artery. Operative mortality was 2.6% (1 patient). There was no observable reversible myocardial ischemia on the postoperative thallium single photon emission computed tomography study. Myocardial revascularization with the use of a skeletonized free right ITA as a composite graft to the left anterior descending artery is an alternative option in cases when an in situ ITA cannot be used. PMID- 12643434 TI - Atrial fibrillation: current surgical options and their assessment. AB - Atrial fibrillation (AF) is the most common cardiac arrhythmia and is associated with morbidity and mortality. Traditional surgical treatment of AF is the Cox Maze III procedure, a complicated operation. New surgical approaches include alternate energy sources (radiofrequency, microwave, cryothermy) and simplified left atrial lesion sets. These operations cure AF in 70% to 80% of patients. This review describes contemporary and emerging surgical approaches to AF, synthesizes results of these operations, and proposes new standards for reporting results of AF treatment. PMID- 12643435 TI - Poland's syndrome revisited. AB - Poland's syndrome is a rare congenital anomaly characterized by unilateral chest wall hypoplasia and ipsilateral hand abnormalities. Literary data suggest its sporadic nature. The prevailing theory of its cause is hypoplasia of the subclavian artery or its branches, which may lead to a range of developmental changes. The incidence of Poland's syndrome varies between groups (male versus female patients, congenital versus familial cases, and so on) and ranges from 1 in 7,000 to 1 in 100,000 live births. Cases of Poland's syndrome associated with leukemia, carcinoma of the hypoplastic breast, and other conditions, confirm the relationship between developmental defects and tumors, and require oncologic awareness. Various manifestations, age, and gender require different surgical approaches. Our experience, which includes 27 patients (15 male, 12 female), 20 of whom (12 male, 8 female) underwent operation, suggests that the repair should be done in two stages in children and in a single stage in adults. Reconstruction and/or stabilization of the aplastic ribs may be achieved using bone grafts or prosthetic mesh. Muscle flaps and breast implants may be used to correct muscle deficiency and breast hypoplasia and to help achieve a complete cosmetic repair. PMID- 12643436 TI - Long-term follow-up of asphyxiating thoracic dystrophy: results of first report of thoracoplasty. PMID- 12643437 TI - Median sternotomy for combined cardiac and descending thoracic aortic operation. PMID- 12643438 TI - Intraarterial thrombolysis for perioperative stroke in patients undergoing cardiac operations. PMID- 12643439 TI - Fontan operation in a patient with separate hepatic venous drainage: is there a problem? PMID- 12643440 TI - Strong LFA-1 and VCAM-1 expression in histological type II of rheumatoid arthritis. AB - With an own histological classification of rheumatoid arthritis (RA) in synovial membranes (SM) of two main types: type I (B-lymphocytic and plasma cellular, local non-destructive, better prognosis); type II (T-lymphocytic, macrophacytic, local destructive, worse prognosis) and type III as a mixed one we examined whether there is a relation between special adhesion molecules and any of this histological types. 32 fresh cryo-conserved RA-SM (type I, II, III; n = 9, 11, 12, respectively) were investigated immunohistochemically using the APAAP method in order to obtain the expression of LFA-1, VCAM-1, CD44 and E-selectin. Positive cells were counted morphometrically within six histological areas: lining layer, subintimal, perivascular, lymphatic follicles, perifollicular and interstitial. Type II showed a significant higher expression than type I for LFA-1 in lining layer and subintimal II (65%; 53% vs 0%; 32%); for VCAM-1 in subintimal, perifollicular, perivascular and interstitial areas (61%, 54%, 58%, 61% vs 6%, 8%, 5%, 6%). In lining layer and lymphatic follicles no significant difference between both types was detected. CD44 and E-selectin: No statistical differences could be found. RA-SM type II shows high expression of LFA-1 and VCAM-1, this is related to a higher destructive process. PMID- 12643441 TI - Ceramide attenuates high glucose-induced cardiac contractile abnormalities in cultured adult rat ventricular myocytes. AB - Ceramide, the metabolic product of signaling molecule sphingomyelin, has been implicated in cardiac Ca2+ regulation. To study the possible role of ceramide in the pathogenesis of diabetic cardiomyopathy, we examined the effects of ceramide on the cardiac contractility of cultured ventricular myocytes under control and simulated diabetic environments. Adult rat ventricular myocytes were maintained in normal (NG, 5.5 mM) or high glucose (HG, 25.5 mM) medium for 24 hr in the absence or presence of the membrane-permeant ceramide analog C2-ceramide, ceramide glucosyltransferase inhibitor D,L-threo-1-pheny-2-decanoylamino-3 morpholino-1-propanol (PDMP), or the inactive ceramide analog C2-dihydroceramide. Contractile indices analyzed included peak shortening (PS), time-to-PS (TPS), time-to-90% relengthening (TR90), maximal velocity of shortening/relengthening (+/- dL/dt), and intracellular Ca2+ fura-2 fluorescence intensity (FFI). Myocytes maintained in HG medium displayed reduced PS and +/- dL/dt associated with prolonged TR90 and normal TPS compared to NG myocytes. Interestingly, the HG induced mechanical dysfunctions were significantly attenuated by C2-ceramide or PDMP. C2-ceramide did not affect the myocyte mechanics in NG myocytes although PDMP shortened TPS without affecting any other indices. The HG-induced contractile abnormalities were not altered by inactive ceramide analog C2 dihydroceramide (except +/- dL/dt). Fura-2 fluorescence recording revealed that HG reduced baseline as well as stimulated intracellular Ca2+ levels, which may be abolished by both C2-ceramide and PDMP. These data suggest that alteration of ceramide signaling may play a role in the pathogenesis of diabetic cardiomyopathy. PMID- 12643442 TI - Fewer beta-endorphin expressing arcuate nucleus neurons and reduced beta endorphinergic innervation of paraventricular neurons in schizophrenics and patients with depression. AB - In order to elucidate whether the hypothalamic expression of beta-endorphin is altered in patients with mental disorders we studied the cellular localization of the peptide in arcuate nucleus neurons as well as the beta-endorphinergic innervation of paraventricular neurons in nine schizophrenics, six subjects with depression, and nine controls. A polyclonal antiserum against beta-endorphin was employed for the immunohistochemical detection of the peptide in sections of postmortem human brains. Quantitative analysis revealed that the number of beta endorphin-containing arcuate neurons was statistically reduced in schizophrenics and depressives in comparison to controls. Moreover, the number of endorphinoceptive (i.e. beta-endorphin-innervated) paraventricular nerve cells was also lower in psychiatric patients than in control cases. Our results showing an altered endorphinergic system in human hypothalami of schizophrenics and depressives might contribute to a renewal of interest in this peptide as a possible factor of importance in psychiatric disorders. PMID- 12643443 TI - In vitro antioxidant properties of calpain inhibitors: leupeptin and calpain inhibitor-1. AB - The aim of this study was 1/ to assess the efficacy of calpain inhibitors: leupeptin and calpain inhibitor-1, to inhibit calpain activity in vitro, 2/ to measure the scavenging abilities of these compounds against free radicals. The efficacy of calpain inhibitors to block calpain activity was tested with azocasein as a substrate for calpain. Leupeptin and calpain inhibitor-1 inhibited calpain activity in the same range of concentrations, the IC50 being 0.14 microM and 0.09 microM, respectively. We measured the antioxidant properties of leupeptin and calpain inhibitor-1 using the allophycocyans assay after identification offree radical species produced by the complex H2O2 + Cu(++). Electron paramagnetic resonance (EPR) spin-trapping studies performed by using DMPO showed that a quartet signal (hyperfine couplings aN = aH = 14.9 G) arisen from DMPO-OH was formed. We found a correlation between leupeptin concentration and oxygen radical absorbance capacity (ORAC) values (r2 = 0.975) indicating an in vivo antioxidant capacity. In contrast, calpain inhibitor-1 showed no protection. In conclusion, our findings indicate that leupeptin and calpain inhibitor-1 are equipotent inhibitors on calpain activity but exhibit diffrent antioxidant efficacy. PMID- 12643444 TI - Interphase fish analysis of cell cycle genes in asbestos-treated human mesothelial cells (HMC), SV40-transformed HMC (MeT-5A) and mesothelioma cells (COLO). AB - The epidemiologic association between asbestos exposure and human malignant mesothelioma is well established. However, the molecular mechanisms linking asbestos exposure of humans and the subsequent mesothelioma formation is not well understood. The most frequent genetic changes found so far in human malignant mesothelioma (HMM) are deletions and point mutations in the tumor suppressor genes p16INK4a and NF2. Whereas homozygous deletions appear to be the predominant mechanism leading to p16/CDKN2A inactivation, inactivating point mutations coupled with allelic loss mainly occur at the NF2 locus. In the present study, asbestos-treated human mesothelial cells (HMC), SV40-transformed human mesothelial cells (MeT-5A) and a human mesothelioma cell line (COLO) were investigated for genetic changes of cell cycle genes (cyclin D1, p16INK4a, RB1, CDK2) using multicolor fluorescence in situ hybridization (mFISH) in interphase cells. The results show that cyclin D1 is unaffected in all investigated cells. The p16INK4a gene locus was shown to be mutated in COLO cells but not in HMC. After labeling of CDK2 and RB1, hemizygous loss of one allele of each gene was observed in asbestos-treated HMC whereas gene amplification of these genes was detectable in MeT-5A and COLO cells. Our data indicate that disarrangement of the RB1 dependent pathway seems to be involved in mesothelioma formation. PMID- 12643445 TI - Inhibition of DNA replication by berenil of bacterial plasmids containing poly(dA)-poly(dT) sequences. 2D gel analysis of replicative intermediates. AB - Berenil, an aromatic compound used in veterinary medicine to treat trypanosome infections in livestock, has been shown to interfere with kinetoplast DNA replication. The drug is thought to bind to the minor groove of DNA and form hydrogen bonds between opposite A/T pairs. Studies utilizing Trypanosoma cruzi, revealed that minicircle DNA, which is 60% A-T rich, and also the major component of kinetoplast DNA networks, is one of the targets for berenil. In order to better understand the mode of action of berenil and its effect on DNA replication, we have studied the effect of the drug on pBR322 derived plasmids containing poly(dA)-poly(dT) sequences. The resulting plasmids were pVL26, which contained 240 bp of poly(dA)-poly(dT) inserted at the EcoRV site of pBR322 and pKH47, which contained 100 bp of poly(dA)-poly(dT) inserted at the PvuII site of pBR322. When cultures containing all of these plasmids were exposed to berenil, plasmids pVL26 and pKH47 were found to have significantly lower yields than pBR322, with pKH47 being the most sensitive to berenil. In the present study we show that the poly(dA)-poly(dT) sequences in plasmids pVL26 and pKH47 are not very stably maintained. However, the resulting deletion mutants containing a fraction of the poly(dA)-poly(dT) sequences were still sensitive to berenil. We also analyzed by 2D agarose gel electrophoresis the progression of the replication fork through the homopolymer region in plasmid pVL26d but failed to detect a replication barrier in this region in the presence of berenil. PMID- 12643446 TI - Comparison of expression patterns of fibroblast growth factor 8, bone morphogenetic protein 4 and sonic hedgehog in jaw development of the house shrew, Suncus murinus. AB - To elucidate the mechanism underlying jaw development in mammals, we used a new laboratory animal, Suncus murinus (house shrew, an insectivore) as the subject for the investigation, because Suncus has all types of teeth (incisor, canine, premolar and molar) in its upper and lower jaws and is thought to be a good model animal having a general mammalian tooth pattern. At the start, by use of degenerate primers we cloned Suncus homologues of fibroblast growth factor 8 (sFgf8), bone morphogenetic protein 4 (sBmp4) and sonic hedgehog (sShh) genes from cDNA library derived from whole Suncus embryos at day 12 (E12). Thereafter, we examined the expression patterns of these genes in the jaw development of Suncus E11-16 embryos (for mouse E9.5-12 embryos). sFgf8 and sBmp4 were expressed in E11 but not in E15 and onward during orofacial development. sShh was expressed from E11 onward, and its expression was increased in the orofacial area. The expression pattern of sFgf8 in the maxillary and mandibular arches of E14 coincided with the area of the presumptive tooth arch. However, sShh and sBmp4 were expressed only in the outer area (= buccal/labial side) of presumptive tooth arch. Thus, these 3 genes showed specific expression pattern in jaw development of Suncus, and their distributions did not overlap each other except in a few regions. These findings suggest that sFgf8, sBmp4 and sShh have a specific function respectively during jaw development in Suncus murinus. PMID- 12643447 TI - Cytochemical demonstration of expression and distribution of non-glycosylated human lysosomal cathepsin S in HEK 293 cells. AB - The lysosomal cysteine protease cathepsin S is synthesized as inactive precursor at the rough endoplasmic reticulum (ER), further processed in the Golgi compartment and finally targeted to the lysosomes where it becomes activated by the proteolytic cleavage of the inhibitory propeptide. Biochemical studies with a non-glycosylated mutant of procathepsin S (plasma membrane binding at 2 degrees C, reuptake of secreted enzyme at 37 degrees C) led to the suggestion of an additional sorting motif in procathepsin S besides the classical Man-6-P recognition signal. In order to further confirm this suggestion on a morphological basis we performed a series of laser scanning confocal microscopy (CLSM) and electron microscopical analyses with HEK 293 cells expressing the mutant non-glycosylated procathepsin S. Immunolocalization with CLSM documented clearly a fine granular fluorescence in the paranuclear region of mutant expressing cells. Electron microscopy demonstrated the presence of cathepsin S immunoreactive deposits within cytosolic vacuoles (lysosomes), at the plasma membrane and in ER buds. These buds were also visible in the cytosol as well as in form of concentrated patches at the plasma membrane indicating the direct transport of (pro)cathepsin S from the ER to the cell surface. PMID- 12643448 TI - Host differences in serum antibody response during infection of goats by caprine arthritis-encephalitis virus. AB - The response of goats infected with caprine arthritis encephalitis virus was evaluated by estimating the binding antibodies in serum. The immunoreactive proteins of the CAEV, strain 63, were determined by immunobloting the lysate against the serum of the infected goat. The major proteins identified were of the molecular weights of 135, 120, 56, 44, 38, 28, 20 and 13 kDa. Proteins with less prominence were of the molecular weights of 96, 32 and 30 kDa. The responses to the viral proteins at several stages of the infection were examined. Antibodies to viral proteins of 135, 121 and 90 kDa were the first to appear in the goat kids that were apparently infected through breast-feeding by infected dam. The time of appearance of these antibodies in the serum varied widely between the goats from one to six months, and preceded the appearence of antibodies to other proteins by two to four weeks. Salient variations revealed in the immunoblot patterns reflect the differences in the immunological responses by the goats to the viral proteins. Littermate goat kids infected through the same dam showed slightly different responses. In the same manner, dissimilarities were observed in the patterns produced by the serums of the damand the corresponding offsprings, when run against the same virus lysate. This study indicates that some differences occur in the immunoblot pattern when this assay is used for diagnosis, although such discrepancies should not affect the interpretation of the diagnostic assay. The host differences in the response to the viral proteins also suggest the potential problems that can be encountered developing an effective vaccine for CAEV, if the binding antibody production reflects that of the neutralizing antibodies in the goats. For effective vaccine coverage, there should be little variability in the immunogenicity of candidate vaccine antigens among the hosts. PMID- 12643449 TI - Evidence of K and Ca channels in endothelial cells of human allantochorial placental vessels. AB - Presence of ionic channels was evaluated in human endothelial cells (ECs) from allantochorial placental vessels by the measure of the membrane potential, the main factor of the excitation-contraction coupling. To characterize endothelial cell channels, vasoactive and vasorelaxant agents and channels blockers were used. In predepolarized cells by high-K+ concentration, the voltage-dependent K+ channels (Kdf) were blocked. Acetylcholine, sodium nitroprusside and isoproterenol hyperpolarized the predepolarized-high K+ ECs. This hyperpolarization was inibited by charybdotoxin (a blocker of big Ca(2+) activated K+ channels) or/and by tetraethylammonium and was insensitive to glibenclamide (a blocker of ATP-sensitive K+ channels). Serotonin depolarized ECs and the depolarization was inibited by nifedipine, a blocker of voltage-gated Ca2+ channels. These results indicated that membrane potential of human placental allantochorial ECs was regulated by voltage-gated, C(2+)-sensitive K+ channels and by voltage-gated Ca2+ channels. PMID- 12643450 TI - Leptin, leptin resistance and endothelial dysfunction in pre-eclampsia. AB - Pre-eclampsia, or pregnancy-induced hypertension, is a major cause of maternal and fetal morbidity and mortality complicating 7-10% of pregnancies. Although mechanisms underlying pre-eclampsia are not well understood, endothelial dysfunction is considered to underscore many of the pre-eclamptic manifestations including hypertension, proteinuria and edema. Leptin, the obese gene product from adipocytes, is produced by the placenta during pregnancy. Serum leptin levels are elevated under normal pregnancy especially during the second trimester, which rapidly decreases and returns to normal after delivery, indicating the role of leptin as a gestational hormone for energy balance. Recent studies have revealed that the placental production of leptin may be pathologically augmented under pre-eclampsia although conflicting data also have been reported. Nevertheless, hyperleptinemia and possible further augmentation under pre-eclampsia may predispose to the development of maternal leptin resistance, which may be a component of insulin resistance predisposing the onset of endothelial dysfunction. This review summarizes recent findings regarding the putative changes of serum leptin levels during pre-eclampsia and the potential consequences on the vascular system. PMID- 12643451 TI - Identification of genes required for immortalization in human papillomavirus infected human oral keratinocytes. AB - The early stages of head and neck cancer are presumed to require a senes of genetic alterations that are not represented by a distinct clinical phenotype. Therefore, genes with altered expression in the preneoplasia may be useful for the early detection of this highly recurrent cancer. In this study, we immortalized normal human oral keratinocytes (NHOK) by retroviral-mediated infection of HPV 16 transforming oncogenes, E6 and E7 (HOK16E6E7). Using the Affymetrix gene chip (U95Av2), we identified 177 known genes and EST that were overexpressed at least 3-fold or above in the immortalized cells, while 133 were down-regulated compared to NHOK. Northern blot analysis showed elevated levels of p55CDC in the immortalized cells, while NHOK showed high basal expression of small proline rich protein (SPRR2). The altered expression of these genes maybe associated with cellular proliferation or differentiation and the early stages of oral carcinogenesis. PMID- 12643452 TI - Jenfluor ap--a novel fluorogenic substrate for in situ detection of alkaline phosphatase activity. AB - Alkaline phosphatase (AP) activity is often targeted in enzyme-related histochemistry as probe enzyme to detect neoplastic cells, as marker for primordial germ cells as well as in preimplantation studies, osteoblast differentiation, phosphate starvation in bacteria, yeast and phytoplankton. Moreover, AP-marker activity is a very useful tool in immunohistochemistry to detect gene sequences, antigens and antibodies. Here we describe a novel high resolution fluorescence method to localize AP-activity in cells and tissue sections based on a naphthol-AS azo coupling procedure (Jenfluor ap). This method provides amorphous photostable fluorescent final reaction products without any diffusion artifacts which are visible in conventional fluorescence microscopes as well as in confocal laser scanning and near infrared multiphoton laser scanning microscopes. The superiority of the Jenfluor ap method in comparison to the known Fast Red TR salt as well as the ELF stains is discussed. PMID- 12643453 TI - Variations in the glycosaminoglycan content, swelling properties and morphological aspects of different regions of the superficial digital flexor tendon of pigs. AB - The superficial digital flexor tendon of pigs is a wrap around tendon supporting compressive in addition to the tensional forces in its proximal region, and only tensional forces in the intermediate region. Clear distinctions were observed in the physical properties, composition of proteoglycan and morphology for the different regions. A greater swelling in water and a larger amount of glycosaminoglycan were found in the proximal region compared to the intermediate one. Dermatan sulfate was detected in all regions of this tendon, while chondroitin sulfate was prominent only near the bone. In the toluidine blue stained sections, intense metachromasy was observed in the sites under compressive forces, especially in the portion close to the bone. Chondrocyte like cells were also observed in these arms. Crimp morphology, observed in the intermediate region, exhibited a clearly sloped aspect in relation to the main axis of the tendon. Elastic fibers were found in all regions, and were disposed in different directions in the areas under compressive forces, and parallel to the collagen bundles in the region under tension. These results reinforce the idea that mechanical forces contribute to a differentiated composition and organization of the extracellular matrix of tendons. PMID- 12643454 TI - Inhibition of 6A8 alpha-mannosidase causes oncosis-like death of BJAB cells. AB - To study relationships between glycosylation and cell survival we used a human alpha-mannosidase, 6A8, recently discovered in our laboratory. We found that glycosylation of BJAB cells was modified when cells were transduced with a fragment of antisense 6A8 cDNA, which resulted in an inhibition of 6A8 alpha mannosidase expression. In comparison to wild type, mock- or sense 6A8-tansduced cells, the antisense 6A8-transduced cells showed increased cell death in culture. The dying cells were characterized by cell and nucleus swelling, membrane blebbing, organelle dilation and membrane breaking at the blebbing sites. They did not have apoptotic bodies. Annexin-V staining and DNA fragmentation assays also showed no evidence of apoptosis in these cells. This data implied that BJAB cells underwent an oncosis-like cell death upon transduction with a fragment of antisense 6A8 cDNA, which inhibited 6A8 alpha-mannosidase expression and therefore modified glycosylation of proteins shown by Con A binding test. These data imply that 6A8 alpha-mannosidase plays an important role in preventing oncosis-like death in BJAB cells. The possible mechanism of cell death by oncosis was discussed. PMID- 12643455 TI - Pinteraction of sex steroid hormones with the eye. AB - Many epidemiologic and laboratory studies in the recent past suggested that the physiology of the eye may be affected by alterations in sex steroid homeostasis due to ageing and pathogenesis. This hypothesis is again strengthened by the presence of specific receptors for steroid hormones in various ocular tissues. However their involvement in pathological process is not yet clearly established. This review deals with a viewpoint on the possible beneficial and detrimental effects of sex steroids on ocular physiopathology. Since sex steroids are different in males and females, an interesting concept of sexual dimorphism in human eye also emerges from this information. PMID- 12643457 TI - Vasopressin therapy for vasoplegic syndrome following cardiopulmonary bypass. AB - BACKGROUND: Hypotension refractory to maximal doses of alpha-adrenergic drugs after cardiac operations employing cardiopulmonary bypass (CPB) has been referred as "vasoplegic syndrome." Vasopressin has been used for its therapy with encouraging results. MATERIAL AND METHODS: 16 patients (mean age 71, range 47 to 84 years) were treated with intravenous vasopressin (0.1-1 IU/min) for hypotension refractory to maximal doses (>30 microg/kg/min) of norepinephrine after undergoing complex cardiac operations employing CPB. Preoperative ejection fraction was 40.5% (mean, range 20% to 60%), preoperative NYHA class was 3.5 (mean). Hemodynamic measurements were obtained one hour before and one hour after beginning vasopressin infusion; urine output was measured for the 4 hours before and the 4 hours after beginning the infusion. Duration of vasopressin treatment was 58.8 +/- 37.3 hours (mean +/- SD). RESULTS: Systolic blood pressure increased from 89.6 +/- 7.9 to 119.6 +/- 10.5 mmHg (mean +/- SD) (p < 0.001); systemic vascular resistance increased from 688.0 +/- 261.7 to 1043.3 +/- 337.1 dyne/s/cm2 (mean +/- SD) (p < 0.001); cardiac index decreased from 2.69 +/- 0.8 to 2.2 +/- 0.5 L/min/m2 (mean +/- SD) (p < 0.008); urine output increased from 36.8 +/- 30.4 to 72.8 +/- 38.2 mL/h (mean +/- SD) (p < 0.001). Seven patients (44%) survived the hospital stay. CONCLUSIONS: High-dose vasopressin is effective in the treatment of the vasoplegic syndrome after cardiac operations employing cardiopulmonary bypass. PMID- 12643458 TI - The results of probe technique for transatrial repair of tetralogy of Fallot. AB - OBJECTIVE: Total correction of classic tetralogy of Fallot (TOF) by transatrial approach has become a standard procedure with a principal theoretical aim of minimizing structural damage to the pulmonary pump. The most critical point in transatrial repair of TOF is infundibular dissection. Right atrial approach provides better surgical exposure for parietal extension of the infundibular septum when compared to a right ventricular approach. However, it is not always easy to determine the localization and amount of muscle bundles to be resected and this surgical maneuver requires experience. METHODS: Nineteen patients were reviewed who had repair of isolated TOF by this technique from 1993 to 2001. The mean age of patients was 5 +/- 2 years. Transatrial-transpulmonary approach was performed for all patients. To make the infundibular muscle-bundle resection easier and to determine the localization and amount of muscle bundle to be resected, we placed a Hegar dilator into the right ventricle through pulmonary arteriotomy. The muscle bundles between the dilator and the anterior leaflet annulus of the tricuspid valve were totally excised until the intraventricular part of the dilator and pulmonary annulus became completely visible. The area between the Hegar dilator and the margins of the ventricular septal defect (VSD) was left untouched. None of the patients had transannular patch. Tricuspid valve detachment in order to improve the exposure was done in 11 patients. All patients were followed up in our clinic at regular six-month intervals by echocardiography. RESULTS: There was no early or late mortality nor reoperation for residual VSD or residual right ventricle (RV) outflow obstruction. All patients were in NYHA class I. RV on the echocardiography was spared late dilatation and had a good late functional status. Eighteen patients had no or mild pulmonary regurgitation. One patient who had undergone tricuspid anterior leaflet detachment showed mild tricuspid insufficiency. CONCLUSIONS: On the basis of hemodynamic outcomes, this procedure for elective repair of TOF in selected cases gives excellent early and mid-term results. PMID- 12643456 TI - Improvement of bypass circuit biocompatibility: comparison and combination of heparin-coated circuit and nitric oxide gas infusion. AB - OBJECTIVES: Nitric oxide (NO) gas infusion to the oxygenator, as well as heparin coated bypass circuits, have been reported to attenuate blood activation induced by the interaction with the artificial surfaces of an extracorporeal bypass circuit. Using a mock circulation model, we compared the effect of each and also evaluated the effect of their combination on attenuating bypass-induced blood activation. METHODS: A miniature closed bypass circuit was primed with diluted fresh human blood and perfused for 180 minutes using a centrifugal pump. NO gas (0, 50, or 100 ppm) was infused to the oxygenator sweep gas of either a non heparin-coated or a heparin-coated circuit. Platelet counts, beta thromboglobulin, platelet factor 4, complement-3 activation products and granulocyte elastase were measured at 0, 30, 60, 120, and 180 minutes after starting the perfusion. RESULTS: One hundred ppm of NO was statistically equivalent to the heparin-coated circuit for attenuating bypass-induced blood activation, and a combination of the two significantly surpassed the results of either modification alone. Fifty ppm of NO alone provided only a slight attenuation of blood activation as compared with the non-heparin-coated circuit, though the difference was not significant. A combination of 50 ppm NO and the heparin-coated circuit did not significantly enhance the effects of the heparin coated circuit alone. CONCLUSIONS: The combination of NO gas infusion and heparin coated circuits appears to be a useful and promising modification for enhancing the attenuation of bypass-induced blood activation, though the optimal dose of NO infusion in terms of effectiveness and adverse effects to the whole body remains to be established. PMID- 12643459 TI - Emergency off-pump coronary artery bypass (OPCAB) for left main coronary occlusion using rapid aorto-coronary perfusion. AB - 64 year old female with occluded left main after first injection brought to the operating room in extremis on IABP and maximum inotropes. LAD grafted with sapenous vein and immediate aorto-coronary perfusion. Circumflex grafted and patient taken to ICU. Patient discharged on POD #6 after echo shows normal ventricular with no wall motion abnormality. PMID- 12643460 TI - Association of sternal wound infection with parasternal muscle sutures. AB - BACKGROUND: Sternal wound infection complicating open-heart surgery is a potentially devastating complication that has been associated with a number of risk factors. We recently consulted on three consecutive patients with this complication who had heavy nonabsorbable parasternal sutures placed in muscle tissue adjacent to the sternum. The aim of this report is to document our findings and caution that this technique to control bleeding from the parasternal intercostal muscles my increase risk of infection. METHODS: The pathology, surgical findings, and microbiology of these three cases are analyzed for similarity and possible cause of infection. RESULTS: By surgical observation and culture reports, each infection appeared to have originated at the site of nonabsorbable suture in devascularized parasternal muscle tissue. Sinus tracts could be probed to a similar site in each patient. CONCLUSION: Placement of sutures in the parasternal muscles where the sternal wires wrap around the bone leads to compression and necrosis of muscle tissue. We caution that this technique to control bleeding may cause a nidus of infection and increase the risk of deep sternal wound infection. PMID- 12643462 TI - Evolution of OPCAB surgery in a single institution. AB - Off-pump coronary artery bypass (OPCAB) grafting is becoming popular worldwide. The development of exposure and stabilization techniques have made this surgery a simple, safe, and routine procedure. The evolution of OPCAB surgery at our institution is presented and discussed. PMID- 12643461 TI - Direct fibrin injection to promote new collateral growth in hind limb ischemia in a rabbit model. AB - Local stimulation of angiogenesis is a new approach for the treatment of critical limb ischemia. Our investigation tested intramuscular (i.m.) injection of a modified fibrin meshwork in a rabbit model. METHODS: The left external iliac and femoral arteries were excised in 24 rabbits that were divided into four groups: control; i.m. saline injection; fibrin meshwork plus low dose (2.5 mg) fibrinogen i.m.; fibrin meshwork plus high-dose (5.0 mg) fibrinogen i.m. Angiography was performed before surgery, immediately after surgery, and one month postoperatively. Lower limb-calf blood pressure was measured immediately after surgery and at postoperative days 10, 20, and 30. On day 30, conventional indirect immunostaining was performed to determine the percentage of the area occupied by capillaries. RESULTS: Immediately after surgery, in all four groups, the number of contract-opacified arteries (COA) crossing a specific segment of a grid decreased from 5.3 +/- 1.3 to 3.2 +/- 1.0 (p < 0.05); the number of grid intersections decreased from 30.2 +/- 6.5 to 19.3 +/- 4.8 (p < 0.05); and the total number of grids with COA decreased from 18.3 +/- 3.8 to 12.2 +/- 2.5 (p < 0.05). One month after surgery, in the control group, these parameters were 6.2 +/- 1.1, 33.2 +/- 5.7 and 20.3 +/- 1.5, respectively; in the saline-treated group, these parameters were 6.1 +/- 0.8, 28.3 +/- 6.9 and 19.8 +/- 1.1, respectively (p > 0.05 versus control and versus baseline data). When fibrin containing 5.0 mg fibrinogen was used, these parameters increased to 8.5 +/- 0.9, 48.3 +/- 5.1, and 27.1 +/- 0.9, respectively (p < 0.001 versus immediately after surgery and p < 0.05 versus control). In all four series, no Doppler flow signal was detected from the posterior tibial artery by day 10. By day 30, the lower limb-calf blood pressure ratio had improved in all four series, but was significantly improved in only the two groups treated with fibrin sealant (0.3 +/ 0.05 control; 0.3 +/- 0.08 saline; 0.6 +/- 0.06 fibrinogen 2.5; 0.7 +/- 0.05 fibrinogen 5.0). CONCLUSION: Intramuscular injection of a fibrin meshwork considerably increased angiogenesis in the severely ischemic hind limb and may be strongly recommended for clinical use in patients with limb-threatening ischemia. PMID- 12643463 TI - Effect of off-pump coronary artery bypass grafting on risk-adjusted and cumulative sum failure outcomes after coronary artery surgery. AB - BACKGROUND AND AIM: We have shown that cumulative sum (CUSUM) failure analysis may be more sensitive than standard statistical methods in detecting a cluster of adverse patient outcomes after cardiac surgical procedures. We therefore applied CUSUM, as well as standard statistical techniques, to analyze a surgeon's experience with off-pump coronary artery bypass grafting (OPCAB) and on-pump procedures to determine whether the two techniques have similar or different outcomes. METHODS: In 320 patients undergoing nonemergent, first time coronary artery bypass grafting, preoperative patient characteristics, rates of mortality and major complications, and ICU and hospital lengths of stay were compared between the on-pump and OPCAB cohorts using Fisher's exact tests and Wilcoxon two sample tests. Predicted mortality and length of stay were determined using previously validated models of the Cardiac Care Network of Ontario. Observed versus expected ratios of both variables were calculated for the two types of procedures. Furthermore, CUSUM curves were constructed for the on-pump and OPCAB cohorts. A multivariable analysis of the predictors of hospital length of stay was also performed to determine whether the type of coronary artery bypass procedure had an independent impact on this variable. RESULTS: The predicted mortality risk and predicted hospital length of stay were almost identical in the 208 on-pump patients (2.2 +/- 3.9%; 8.2 +/- 2.5 days) and the 112 OPCAB patients (2.0 +/- 2.2%; 7.8 +/- 2.1 days). The incidence of hospital mortality and postoperative stroke were 2.9% and 2.4% in on-pump patients versus zero in OPCAB patients (p = 0.09 and 0.17, respectively). Mechanical ventilation for greater than 48 hours was significantly less common in OPCAB (1.8%) than in on-pump patients (7.7%, p = 0.04). The rate of 10 major complications was 14.9% in on pump versus 8.0% in OPCAB patients (p = 0.08). OPCAB patients experienced a hospital length of stay that was a median of 1.0 day shorter than on-pump patients (p = 0.01). The observed versus expected ratio for length of stay was 0.78 in OPCAB patients versus 0.95 in on-pump patients. On CUSUM analysis, the failure curve in OPCAB patients was negative and was flatter than that of on-pump patients throughout the duration of the study. Furthermore, OPCAB was an independent predictor of a reduced hospital length of stay on multivariable analysis. CONCLUSIONS: OPCAB was associated with better outcomes than on-pump coronary artery bypass despite a similar predicted risk. This robust finding was documented on sensitive CUSUM analysis, using standard statistical techniques and on a multivariable analysis of the independent predictors of hospital length of stay. PMID- 12643464 TI - Systematic organ protection in coronary artery surgery with or without cardiopulmonary bypass. AB - Coronary artery bypass grafting with cardiopulmonary bypass (CPB) is known to be associated with a systemic inflammatory response, which contributes to the development of postoperative complications including multiple organ dysfunction. Off-pump coronary surgery has been shown to attenuate the inflammatory injury compared to the conventional approach, thereby reducing the incidence of postoperative cardiopulmonary, renal, or neurological dysfunction. It is believed that off-pump experience may greatly impact on improving the outcome of coronary surgery in certain high-risk patients. Moreover, a better understanding of the underlying mechanism would also help to improve our current CPB management. Accumulating evidence to date indicates that a balance between pro- and antiinflammatory responses is crucial in limiting the extent of such systemic inflammatory injury following surgical myocardial revascularization. PMID- 12643465 TI - Ischemic preconditioning: fact or fantasy? AB - Fifteen years ago, an experimental effort to magnify a myocardial infarction, with preinfarction episodes of transient ischemia, proved paradoxically protective. In the ensuing years, surgeons have learned to discriminate a biochemical/metabolic/functional spectrum of cardiac states ranging from healthy myocardium to "stunned" or "hibernating" heart to the modes of "apoptotic" or "necrotic" cardiomyocyte death. It is now clear that "protective cardiac preconditioning" influences all of these cardiac states. The cellular mechanisms of preconditioning (PC) are now sufficiently understood to permit clinical application. Ligation of adrenergic, adenosine, bradykinin or opioid receptors involves signaling via both tyrosine and calcium-dependent protein kinases (PKC), which activate mitochondrial ATP-dependent potassium channels. Subsequently, the release of oxygen radicals induces nuclear translocation of transcriptional regulators, which transform the cardiomyocyte into a more resilient cell. Although preconditioning was initially recognized as protecting only against infarction, PC also limits postischemic dysrhythmias and enhances contractile function. Phase I (safety) and phase II (efficacy) clinical trials now persuasively support pharmacological preconditioning as a safe mode of preventing postcardiac surgical complications. Indeed, preconditioning is currently being proposed as adjunctive to hypothermic perfusates in protecting against the obligate organ ischemia during transplantation. PMID- 12643466 TI - Development and use of gene transfer for treatment of cardiovascular disease. AB - Gene therapy holds promise for the treatment of cardiovascular diseases for which effective pharmacological therapies are insufficient or unavailable. Recent studies have suggested that modification of current gene delivery systems combined with the use of efficacious therapeutic genes may ultimately be successful for clinical vascular gene therapy. Although certain applications such as vein-graft failure may be best suited for short-term transient overexpression of therapeutic genes, other disorders including human essential hypertension and atherosclerosis require sustained overexpression of genes. Hence, design and use of vector systems for delivery of genes to the required site in vivo requires careful consideration. Both viral and nonviral gene therapy vectors show low efficiency for gene transfer into vascular cells and demonstrate a lack of selectivity, as vectors have natural tropism for other cells and tissues. Recent work has focused on the design, development, and utility of vascular cell selective gene therapy vectors for use in distinct and diverse vascular gene therapy scenarios. Using phage display technology we have isolated small peptide ligands that mediate selective binding to either vascular endothelial cells or vascular smooth muscle cells. When engineered into either adenoviral (Ad) or adeno-associated viral (AAV) vectors, candidate peptides enabled the virus to selectively bind to the desired cell type thus generating novel vascular cell selective gene transfer. As preclinical studies have highlighted both the potential for vascular gene therapy as well as defining the potential pitfalls, the development of disease-selective gene therapeutics will increase safety and efficiency of gene therapy for future clinical use. PMID- 12643467 TI - Gene therapy for all aspects of vein-graft disease. AB - Gene therapy could improve human saphenous vein (HSV) coronary vein-graft patency by reducing early thrombosis, neointimal hyperplasia and atherosclerosis. Mouse and rabbit models use veins with much thinner walls than pig or HSVs but atherosclerosis can be more easily induced; none of these models shows early thrombosis. Prostacyclin synthase, tissue factor pathway inhibitor, and tissue plasminogen activator might decrease thrombus formation. Tissue inhibitors of metalloproteinases (TIMPs) reduce intimal migration of smooth muscle cells, while TIMP-3 and the p53 tumor suppressor protein promote apoptosis. Prostacyclin synthase and nitric oxide synthase, and cell cycle inhibitors, such as E2F decoy oligonucleotides (D-E2F), reduce neointima formation. This might be enough by itself to decrease later atherosclerosis. Alternatively, direct targeting with nitric oxide synthase, decoy adhesion molecules, or interleukin-10 might be possible. PMID- 12643468 TI - Does external stenting reduce porcine vein-graft occlusion via an action on vascular nerves? AB - BACKGROUND: Neural reorganization occurs in porcine vein grafts and placement of an external stent reduces graft occlusion. AIM OF THE STUDY: To determine the effect of external stenting on the innervation of porcine vein grafts. METHODS: Saphenous vein into carotid artery grafting (with and without external stents) was performed in 16 pigs. After one and six months, grafts were removed, nerves were counted, and neointima was assessed. RESULTS: In vein graft compared to ungrafted vein, there was a significant (p < 0.05) decrease in medial perivascular nerves, but a dramatic increase in paravascular nerves in the adventitia (p < 0.05). In stented vein grafts there was also a reduction of perivascular nerves and the paravascular nerve proliferation observed in vein grafts at one month was inhibited (p < 0.05). Neointima formation and the appearance of large paravascular nerve bundles in the adventitia of vein grafts were abolished by external stenting. CONCLUSIONS: Neural reorganization plays a role in vein-graft failure, possibly through the local release of mitogens; the prevention of this reorganization contributes to the inhibitory effect of the external stent on neointima formation. PMID- 12643469 TI - Tumor cell hypoxia and the hypoxia-response signaling system as a target for prostate cancer therapy. AB - The accumulation of cancerous cells within a growing prostate tumor can deprive them of adequate vascular support. Without this support, the affected tumor cells become hypoxic, a condition that is usually unfavorable for the further growth and survival of eukaryotic cells. Mammalian cells, however, have the ability of responding to a hypoxic environment by activating a "hypoxia-response" signaling system. Ultimately, this signaling system upregulates the expression of a network of gene products that increase the propensity of the cell to survive even in this adverse environment. With increasing evidence that hypoxia and an activated hypoxia-response signaling system can influence progression (via increased angiogenic propensity and apoptotic resistance) and the therapeutic responsiveness of prostate cancer cells, this review will examine the concept of targeting hypoxia or the hypoxia-response system of prostate tumor cells as a means to suppress prostate tumor progression and metastasis or perhaps even as a means for eliminating prostate tumors in advanced prostate cancer patients. PMID- 12643470 TI - TGF-betal/Smad signaling in prostate cancer. AB - Adenocarcinoma of the prostate is the most common type of cancer, excluding skin cancer, and the second leading cause of cancer death in adult men in the United States. The lifetime risk for developing symptomatic prostate cancer is one in five for an American man. A pivotal step in carcinogenesis is a shift in the balance between proliferation, differentiation, and apoptosis that favors cell proliferation. Transforming growth factor-beta (TGF-beta) is a key negative growth regulator in the normal prostate. Although TGF-beta) inhibits the proliferation of normal prostate cells and functions as a tumor suppressor in early tumorigenesis, it acts as a tumor promoter in later stages of tumor progression. Elevated expression of TGF-beta in prostate cancer cells is associated with poor clinical outcome. Over-expression of TGF-beta aids tumorigenesis by not only stimulating angiogenesis and suppressing the immune system, but also by acting directly on the prostate tumor cells. While prostate cancer cells become resistant to TGF-beta-induced growth inhibition and apoptosis, they retain other TGF-beta-induced responses that enhance tumorgenicity. such as induction of extracellular matrix proteins, cell adhesion proteins and proteases. These direct tumor effects are mediated primarily through Smad signaling. This review addresses the mechanisms by which prostate cancer cells may acquire TGF-beta resistance and promote tumorgenicity. Understanding the mechanisms underlying TGF-beta resistance is important for the identification and development of better diagnostic markers and more effective strategies for treating prostate cancer. PMID- 12643471 TI - Antisense targets to enhance hormone and cytotoxic therapies in advanced prostate cancer. AB - The main obstacle to improved survival of advanced prostate cancer is our failure to prevent or treat its progression to its lethal and untreatable stage of androgen independence. New therapeutic agents designed to prevent androgen independent progression are required. Accelerated identification and characterization of cancer-relevant molecular targets has sparked considerable interest in the development of new generations of anti-cancer agents that specifically inhibit a progression-relevant target. Antisense oligonucleotides, short synthetic stretches of chemically modified DNA capable of specifically hybridizing to the mRNA of a chosen cancer-relevant target gene. promise to show enhanced specificity for malignant cells with a more favorable side-effect profile due to well-defined and tailored modes of action. Although not all of the challenges have been met to date, emerging clinical evidence supports the premise that antisense oligonucleotides stand a realistic chance of emerging as major partners of rationally designed anti-cancer regimens. The status of antisense targeting of several genes, including bcl-2, bcl-xL, clusterin, androgen receptor and IGFBPs, relevant to prostate and other cancers, are reviewed. PMID- 12643472 TI - Par-4 for molecular therapy of prostate cancer. AB - Prostate cancer is the most frequently diagnosed malignancy and the second leading cause of cancer deaths in American men. Although many treatment measures such as androgen deprivation, radiation therapy, and cryoablation exist for primary prostate cancer, there is currently no effective treatment for patients presenting advanced or metastatic stages of the disease. Molecular therapy offers an attractive approach to the treatment of primary prostate cancer because the prostate is not a life-sustaining organ, and a number of tissue specific promoters can be used for prostatic gene expression following relatively straightforward delivery routes. This review discusses the general molecular therapy applications in the context of prostate cancer, and most importantly, identifies the prostate apoptosis response-4 (Par-4) gene, which exclusively induces apoptosis in cancer cells and not normal cells, as a prospective molecule for therapy of the disease. PMID- 12643473 TI - Phytoestrogens and prostate cancer. AB - Androgcns are required to maintain the integrity of the prostate and the survival of androgen dependent epithelial cells within the gland. Anti-androgens arc the primary treatment strategy for non-localized prostate cancer, but ultimately fail over time with the development of androgen independent tumors. Estrogens affect the growth and development of the prostate and may affect the development of prostate cancer. Because of the side effects of estrogen treatment alternative therapies include the use of phytoestrogens as chemopreventative and chemotherapeutic treatment modalities. Phytoestrogens, can cause growth arrest and in some cases apoptosis in prostate cancer cells in vivo and in vitro. This may be due to the estrogenic properties of the compounds or alternative mechanisms of action. A number of phytoestrogens have been shown to have anti androgenic effects and anti-oxidant activities. Other mechanisms include inhibition of 5alpha-reductase, 17beta-hydroxysteroid dehydrogenase, aromatase, tyrosine specific protein kinases and DNA topoisomerase II. This review examines the possible relation between phytoestrogens and prostate cancer and their possible use in prostate cancer prevention or management. PMID- 12643474 TI - From TGF-beta to cancer therapy. AB - This article will introduce a novel concept in the use of TGF-beta insensitive host immune cells in cancer therapy. TGF-beta is a multi-functional cytokine. At a cellular level, it mediates cellular proliferation, growth arrest, differentiation and apoptosis. Because of the above cellular effects, TGF-beta is able to regulate a host of patho-physiological events in vivo, such as normal embryonic development, angiogenesis in tumor tissues, malignant transformation and immune surveillance. As a general rule, its direct effect on cancer cells is inhibition to cancer growth. However cancer cells are able to acquire the ability to evade this inhibitory effect of TGF-beta by becoming insensitive to TGF-beta. Furthermore, these malignant cells are able to produce large quantities of TGF beta. The consequence of over expression of TGF-beta by cancer cells is an important factor for subsequent tumor progression. The excess amount of TGF-beta promotes tumor angiogenesis and immune suppression. The latter effect of TGF-beta is the most devastating to the host. The present discussion is focused on the role of TGF-beta insensitive immune cells in cancer growth. The host immune system offers a natural defense program against cancer. But, this natural immune surveillance is rendered ineffective by an overproduction of TGF-beta derived from the tumor cells. Rendering the host immune cells insensitive to TGF-beta in a gene therapy program offers a hope for us to successfully combat against cancer. Based on the above discussion, it is encouraging that there is a possibility for us to achieve a cure in cancer using TGF-beta insensitive immune cells in gene therapy. PMID- 12643476 TI - Mouse strains for prostate tumorigenesis based on genes altered in human prostate cancer. AB - Animal models of prostate cancer have been limited in number and in relevance to the human disease. With the advancement of transgenic and knockout technologies, combined with tissue specific promoters and tissue-specific gene ablation, a new generation of mouse models has emerged. This review will discuss various animal models and their inherent strengths and weaknesses. A primary emphasis is placed on mouse models that have been designed on the basis of genetic alterations that are frequently found in human prostate cancer. These models display slow, temporal development of increasingly severe histopathologic lesions, which are remarkably restricted to the prostate gland, a property similar to the ageing related progression of this disease in humans. The preneoplastic lesions, akin to what is considered as prostatic intraepithelial neoplasia, are consistent major phenotypes in the models, and, therefore. are discussed for histopathologic criteria that may distinguish their progressions or grades. Finally, considering that prostate cancer is a complex multifocal disease, which is likely to require multiple genetic/epigenetic alterations, many of these models have already been intercrossed to derive mice with compound genetic alterations. It is predicted that these and subsequent compound mutant mice should represent "natural" animal models for investigating the mechanism of development of human prostate diseases, as well as, for preclinical models for testing therapeutics. PMID- 12643475 TI - Therapeutic targets for metastatic prostate cancer. AB - Prostate cancer is the most commonly diagnosed non-cutaneous cancer in adult males. Although prostate cancer that is confined to the gland can be cured in many patients using surgery or radiation, these treatments are only effective for localized tumors and the long-term failure rates for these treatments suggests that prostate cancer can metastasize relatively early in the course of the disease. Once prostate cancer has metastasized there are no curative therapies. The greatest challenge in the treatment of advanced prostate cancer is to access and eliminate metastatic cells. Therefore, effective prostate cancer therapy will require novel strategies to target cancer cells both at the site of the primary tumor and at distant metastatic sites. In this article we review several therapeutic targets and approaches that may provide new treatments for metastatic prostate cancer. We discuss the use of small molecules to target specific molecular events associated with metastatic prostate cancer, the use of specific antibodies that target unique metastasis associated molecules and the use of various gene therapy strategies to achieve anti-metastatic activities. PMID- 12643477 TI - Impact of early postweaning growth rate as affected by diet complexity and space allocation on subsequent growth performance of pigs in a wean-to-finish production system. AB - The objective was to evaluate the effect of restricted early postweaning growth rate due to diet complexity, pen space, or both on subsequent growth to market in a wean-to-finish system. Pigs (n = 1,728) were used in a randomized block design with a 2 x 2 factorial arrangement of treatments: 1) diet complexity (Complex vs Simple) and 2) space allocation (Unrestricted vs Restricted). Treatments were imposed for the first 8 wk after weaning (period 1) and growth was measured from weaning (5.0 +/- 0.01 kg body weight; 15 d of age) to the end of wk 23 postweaning. The Simple diet was based on corn-soybean meal with minimal inclusion of milk products, processed cereals, and animal protein-based ingredients compared to the Complex diet. Floor and feeder-trough spaces were 0.63 m2 and 4 cm and 0.21 m2 and 2 cm per pig for Unrestricted and Restricted space treatments, respectively. From the end of wk 8 to end of wk 23 (period 2), pigs on all treatments had the same floor and feeder spaces and were fed common diets. There was no interaction (P > 0.05) between diet and space treatments. In period 1, Simple diets resulted in similar average daily feed intake (ADFI; 639 vs 650 +/- 5.4 g; P > 0.05), but lower average daily gain (ADG; 408 vs 424 +/- 3.8 g; P < 0.01) and gain:feed ratio (0.64 vs 0.65 +/- 0.002; P < 0.001), and lighter body weight (2.8%; P < 0.01) compared to the Complex diets. In period 2, growth was not affected (P > 0.05) by previous diet complexity, and pig body weight was similar (114.4 vs 114.4 +/- 0.37 kg, P > 0.05) at the end of wk 23. In period 1, pigs with Restricted space had lower ADG (398 vs 434 +/- 3.8 g; P < 0.001), ADFI (621 vs 668 +/- 5.4 g; P < 0.001), and gain:feed ratio (0.64 vs 0.65 +/- 0.002; P < 0.01), and were lighter at the end of wk 8 (6.5%; P < 0.001) than those with Unrestricted space. However, in period 2, pigs with Restricted space had higher (P < 0.01) ADG (3%), ADFI (2%), and gain:feed ratio (3%) than those with Unrestricted space, and body weight was similar (114.5 vs 114.3 +/- 0.37 kg; P > 0.05) at end of wk 23. Carcass backfat and loin-eye depth at market body weight were influenced by neither diet nor space treatment. Using a simple diet program and restricted space allowance immediately postweaning resulted in a lower early growth rate, but had no impact on pig body weight or carcass measures at market. PMID- 12643478 TI - Persistence of carotenoid pigments in the blood of concentrate-finished grazing sheep: its significance for the traceability of grass-feeding. AB - Carotenoid pigments are good biomarkers of grass feeding in sheep. However, grazing lambs are often stall-finished because of grass shortage. We investigated the nature of the carotenoids present in sheep blood and their persistence in this tissue. Four treatments were compared: 1) feeding a concentrate-based diet (n = 10 lambs), 2) grazing followed by a long stall-finishing period (n = 10), 3) grazing followed by a short stall-finishing period (n = 10), and 4) grazing to slaughter weight (n = 10). The concentrate supply was regulated to have similar average daily gain for all treatments. The 40 lambs were allocated to either the grazing or the stall treatments on the basis of their birth date, birth weight, and body weight. The 30 grazing lambs were further allocated to long-stall, short stall, or grass treatment on the basis of their body weight and plasma carotenoid content. Plasma content of total carotenoids was measured by spectrophotometry during the grazing and the stall periods for all lambs and at slaughter weight for the eight heaviest lambs of each treatment. Analysis of the nature and the concentration of individual carotenoids was performed by HPLC on pasture and stall diets and on blood of grazing lambs. The carotenoid content of the stall diet was 2 to 3% that of the pasture diet. Lutein, zeaxanthin, and beta-carotene accounted for 43 to 58%, 3 to 17%, and 0 to 7% of total plasma carotenoids in grazing lambs, respectively. Two unknown polar carotenoids, expressed in lutein equivalent, accounted for 10 to 22% and 0 to 9% of total carotenoids. Plasma carotenoid content during the grazing and the finishing periods varied among animals (P < 0.001). At slaughter weight, plasma carotenoid content was higher for grass-fed than for stall-fed, long-stall finished, or short-stall finished lambs (P < 0.001), and reliably distinguished grass-fed lambs from all the others. Plasma carotenoid content decreased exponentially with the interval from starting on the stall diet (P < 0.005). The deceleration parameter of the model increased linearly with lamb average daily gain during the stall-finishing period, suggesting that the turnover of carotenoids in the blood may depend on the level of intake of the stall-finishing diet. After 4 to 13 d on the stall diet, depending on the initial plasma carotenoid concentration, plasma carotenoid concentration of previously grazed, stall-finished lambs fell to the values of lambs fed a concentrate diet without grazing. Such a low persistence is of interest for discriminating grazing lambs from stall-finished grazing lambs. PMID- 12643479 TI - Evaluation of the South African Dorper as a terminal sire breed for growth, carcass, and palatability characteristics. AB - The South African Dorper is an important terminal meat sire breed in Africa that was recently imported into the United States. The objective of this study was to evaluate the Dorper as a terminal meat sire breed for U.S. production. Semen from purebred Dorper sires was used to artificially inseminate Columbia ewes to produce F1 crossbred lambs. Growth and carcass characteristics of F1 Dorper Columbia lambs (n = 165), F1 Suffolk-Columbia (n = 89), and purebred Columbia lambs (n = 207) were compared based on subsets of the total number of animals. The F1 Dorper lambs grew significantly slower (313 g x d(-1)) and weighed less (29.8 kg) than F1 Suffolk- (357 g x d(-1), 33.5 kg) and Columbia-sired lambs (328 g x d(-1), 31 kg) at 77 d of age. However, at a weaning age of 118 d weaning weight and ADG did not differ (P > 0.20) among sire breed groups. Postweaning growth of F1 Dorper(239 g xd(-1)) wether lambs did not differ from that of purebred Columbia wethers (230 g x d(-1)) but was less than that of F1 Suffolk lambs (259 g x d(-1); P= 0.09). Feed efficiency did not differ among breed types. Breed types had similar dressing percentages (53%), shoulder fat depth (2.8 mm), body wall thickness (3 cm), leg conformation score (Choice), Yield grade (2.4), and Quality grade (Choice). Weight of wholesale shoulders and racks made up approximately 38% of the carcass weight in the Columbia and F1 Suffolk-Columbia type but only 33% in the F1 Dorper-Columbia lambs. However, the more expensive wholesale loins from F1 Dorpers were heavier (P < 0.01) than the other breed types. Total weights of wholesale legs were similar among F1 Dorpers and F1 Suffolks but were heavier than those for the purebred Columbia (P < 0.05). Percentages of total wholesale primal cuts were similar among breed types (P > 0.10). Chemical composition of the carcass did not differ significantly between breed types with a mean composition of 52% moisture, 30% lipid (ether extract), 17% protein, and 0.76% ash. Warner-Bratzler shear force values were less (P < 0.05) and sensory panel ratings for tenderness were significantly more favorable for lamb chops from Dorper sired lambs. Dorper rams can be used as terminal meat sires to produce lambs whose growth rate to 118 d of weaning age, postweaning ADG and feed efficiency, and carcass characteristics are similar to that of Suffolk crossbred lambs and purebred Columbia lambs but with a slight improvement in tenderness. PMID- 12643480 TI - Direct and correlated responses to selection for yearling weight on reproductive performance of Nelore cows. AB - Data from a selection experiment for growth carried out in Brazil were analyzed in order to evaluate the direct responses on yearling weight (YW) and the correlated responses on the size and reproduction traits of cows. The experiment was started in 1976, and in 1980 three lines of Nelore cattle were established: selection (NeS), traditional (NeT), both selected for higher YW, and control (NeC), selected for mean YW. The NeT was an open line that eventually received bulls from other herds. Yearling weight records for animals born from 1978 to 1998 and yearling hip height (H550) offemales born from 1985 to 1998 were analyzed by fitting an animal model in order to obtain the genetic trends. The means for weight, height, and body condition score at the start of the breeding season, days to calving, and calving success of cows born from 1993 to 1996 (pertaining to the third to fourth generations of selection) were compared between the selected (NeS and NeT) and control lines. The genetic trends obtained after 16 yr for YW were 1.7 +/- 0.2, 2.3 +/- 0.2, and -0.1 +/- 0.1 kg/yr for males and 1.9 +/- 0.2, 2.4 +/- 0.2, and -0.1 +/- 0.1 kg/yr for females, for the NeS, NeT, and NeC lines, respectively. Corresponding values for H550 were 0.25 +/ 0.03, 0.24 +/- 0.04 and -0.04 +/- 0.03 cm/yr for females. Heifers and cows from NeS and NeT were 19% and 15% heavier and 4% taller at the start of the breeding season than those from NeC. No significant differences between selected (NeS and NeT) and control females were detected for body condition scores and for reproductive performance. The results indicate that selection for body weight promoted high and consistent weight and height responses both at the yearling and later ages, without compromising the reproductive performance of the cows with respect to days to calving and calving success. PMID- 12643481 TI - Detection of quantitative trait loci for fat androstenone levels in pigs. AB - A QTL analysis of fat androstenone levels from a three-generation experimental cross between Large White and Meishan pig breeds was carried out. A total of 485 F2 males grouped in 24 full-sib families, their 29 parents and 12 grandparents were typed for 137 markers distributed over the entire porcine genome. The F2 male population was measured for fat androstenone levels at 100, 120, 140, and 160 d of age and at slaughter around 80 kg liveweight. Statistical analyses were performed using two interval mapping methods: a line-cross (LC) regression method, which assumes alternative alleles are fixed in founder lines, and a half- full-sib (HFS) maximum likelihood method, where allele substitution effects were estimated within each half- and full-sib family. Both methods revealed genomewide significant gene effects on chromosomes 3, 7, and 14. The QTL explained, respectively, 7 to 11%, 11 to 15%, and 6 to 8% of phenotypic variance. Three additional significant QTL explaining 4 to 7% of variance were detected on chromosomes 4 and 9 using LC method and on chromosome 6 using HFS method. Suggestive QTL were also obtained on chromosomes 2, 10, 11, 13, and 18. Meishan alleles were associated with higher androstenone levels, except on chromosomes 7, 10, and 13, although 10 and 13 additive effects were near zero. The QTL had essentially additive effects, except on chromosomes 4, 10, and 13. No evidence of linked QTL or imprinting effects on androstenone concentration could be found across the entire porcine genome. The steroid chromosome P450 21-hydroxylase (CYP21) and cytochrome P450 cholesterol side chain cleavage subfamily XIA (CYP11A) loci were investigated as possible candidate genes for the chromosome 7 QTL. No mutation of coding sequence has been found for CYP21. Involvement of a candidate regulatory mutation of CYP11A gene proposed by others can be excluded in our animals. PMID- 12643482 TI - Genetic relationships between scrotal circumference and female reproductive traits. AB - Records for yearling scrotal circumference (SC; n = 7,580), age at puberty in heifers (AP; n = 5,292), age at first calving (AFC; n = 4,835), and pregnancy, calving, or weaning status following the first breeding season (PR1, CR1, or WR1, respectively; n = 7,003) from 12 Bos taurus breeds collected at the Meat Animal Research Center (USDA) between 1978 and 1991 were used to estimate genetic parameters. Age at puberty (AP) was defined as age in days at first detected ovulatory estrus. Pregnancy (calving or weaning) status was scored as one for females conceiving (calving or weaning) given exposure during the breeding season and as zero otherwise. The final model for SC included fixed effects of age of dam at breeding (AD), year of breeding (Y), and breed (B) and age in days at measurement as a covariate. Fixed effects in models for AP and AFC were AD, Y, B, and month of birth. Fixed effects in models for PR1, CR1, and WR1 included AD, Y, and B. For all traits, random effects in the model were direct genetic, maternal genetic, maternal permanent environmental, and residual. Analyses for a three trait animal model were carried out with SC, AP, and a third trait (the third trait was AFC, PR1, CR1, or WR1). A derivative-free restricted maximum likelihood algorithm was used to estimate the (co)variance components. Direct and maternal heritability estimates were 0.41 and 0.05 for SC; 0.16 and 0.03 for AP; 0.08 and 0.00 for AFC; 0.14 and 0.02 for PR1; 0.14 and 0.03 for CR1; and 0.12 and 0.01 for WR1. Genetic correlations between direct and maternal genetic effects within trait were -0.26, -0.63, -0.91, -0.79, -0.66, and -0.85 for SC, AP, AFC, PR1, CR1, and WR1, respectively. Direct genetic correlations between SC and AP and between those traits and AFC, PR1, CR1, and WR1 ranged from -0.15 (between SC and AP) to 0.23 (between AP and WR1). Estimates of heritability indicate that yearling SC should respond to direct selection better than AP, AFC, PR1, CR1, and WR1. Variation due to maternal genetic effects was small for all traits. No strong genetic correlations were detected between SC and female reproductive traits or between AP and the other female traits. These results suggest that genetic response in female reproductive traits through sire selection on yearling SC is not expected to be effective. PMID- 12643483 TI - Rendement Napole gene effects and a comparison of glycolytic potential and DNA genotyping for classification of Rendement Napole status in Hampshire-sired pigs. AB - The purpose of the present study was to compare Rendement Napole (RN) classification from glycolytic potential (GP) and DNA techniques, and to study the effect of the RN gene on performance, carcass, muscle quality, and sensory traits. Progeny (N = 118) from the mating of 15 purebred Hampshire sires to York x Landrace females were classified for RN gene status using the GP of the loin and polymerase chain reaction-restriction fragment length polymorphism sequence methodology. Females mated in the study (N = 32) were considered normal (rn+/rn+) based on a loin GP measurement taken on samples collected by live press biopsy. Progeny were randomly selected for harvest within a litter for each sire. Observed mean, standard deviation, and range of progeny loin GP values were 132.2, 30.7, and 70.0 to 193.0 micromol/g, respectively. The GP data were not normally distributed. Peak numbers of observations occurred between 120 and 129 micromol/g and 160 to 169 micromol/g. Pigs with a loin GP of > 150 micromol/g were classified RN-/rn+ based on the observed valley between the peak values, resulting in 37 pigs classified as RN-/rn+ and 81 pigs classified as rn+/rn+. Using DNA procedures, 81 RN-/rn+ and 37 rn+/rn+ pigs were observed. All classification errors occurred when GP values were < or = 150 micromol/g, with 30 of 44 and 14 of 44 classification errors occurring when loin GP values were between 121 and 150 micromol/g and 70 and 120 micromol/g, respectively. Gene effects, based on DNA results, were evaluated using mixed-model procedures with fixed effects of DNA genotype and gender, and random sire and litter effects. No RN genotype differences for growth rate,10th-rib backfat, or loin muscle area were observed. Loins from the RN-/rn+ pigs had significantly (P < 0.05) lower ultimate pH (0.16 units), greater GP (50.3 micromol/g), greater drip loss (0.93%), paler objective color (L*, 1.66 units), paler visual color (0.31 units), and lower firmness (0.61 units) scores. Additionally, loins from RN-/rn+ pigs had significantly (P < 0.05) lower marbling scores (0.68 units) and intramuscular fat content (0.25%) and greater cooking loss (2.51%). Cooked moisture, juiciness score, and mechanical and sensory tenderness measures did not differ between genotypes. The GP-based classification did not correctly classify RN genotype in the present study, emphasizing the importance of the direct DNA analysis for estimation of gene frequency and effects. The DNA-based genotype results clearly indicate the RN- allele has negative effects on muscle quality measures. PMID- 12643485 TI - The effects of ractopamine on the behavior and physiology of finishing pigs. AB - The objectives of this study were to examine the effects of ractopamine (RAC) on the behavior and physiology of pigs during handling and transport. Twenty-four groups of three gilts were randomly assigned to one of two treatments 4 wk before slaughter: finishing feed plus RAC (10 ppm) or finishing feed alone. Pigs were housed in the same building in adjacent pens with fully slatted floors and ad libitum access to feed and water. Behavioral time budgets were determined in six pens per treatment over a single 24-h period during each week. Behavioral responses of these pigs to routine handling and weighing were determined at the start of the trial and at the end of each week. Heart-rate responses to unfamiliar human presence were measured in all pigs and blood samples were taken from a single pig in each pen on different days during wk 4. At the end of wk 4, all pigs were transported for 22 min to processing. Heart rate was recorded from at least one pig per pen during transport and a postmortem blood sample was taken from those pigs that were previously sampled. During wk 1 and 2, RAC pigs spent more time active (P < 0.05), more time alert (P < 0.05), and less time lying in lateral recumbency (P < 0.05). They also spent more time at the feeder in wk 1 (P < 0.05). At the start of the trial, there were no differences in behavioral responses to handling. However, over each of the next 4 wk, fewer RAC pigs exited the home pen voluntarily, they took longer to remove from the home pen, longer to handle into the weighing scale and needed more pats, slaps, and pushes from the handler to enter the scales. At the end of wk 4, RAC pigs had higher heart rates in the presence of an unfamiliar human (P < 0.05) and during transport (P < 0.05), but not during loading and unloading. Also at the end of wk 4, RAC pigs had higher circulating catecholamine concentrations (P < 0.05) than control pigs. Circulating cortisol concentrations and cortisol responses to transport did not differ between treatments. The results show that ractopamine affected behavior, heart rate, and catecholamine profile of finishing pigs and made them more difficult to handle and potentially more susceptible to handling and transport stress. PMID- 12643484 TI - Aggressive behavior is reduced in bulls actively immunized against gonadotropin releasing hormone. AB - The purpose of this research was to compare the frequency of aggressive behavior's in beef bulls actively immunized against gonadotropin-releasing hormone relative to contemporary nonimmunized control bulls and surgically castrated steers. Eight males were assigned to each ofthese treatments in each of 4 yr. Immunized males were treated with a GnRH-keyhole-limpet hemocyanin (KLH) conjugate at approximately 4 mo of age. A secondary (booster) immunization was administered at 12 mo. Steers were castrated at 4 mo of age. Animals in each treatment in each year were housed as a single group prior to testing. At approximately 16 mo of age, each group of eight animals was placed in a 10- x 16 m enclosure for 20 min on five occasions at 2 to 3 d intervals. An observer recorded butts initiated by each animal as well as participation in bouts of sparring. Relative to control bulls, immunocastration reduced the frequency of butts initiated (P < 0.05) and participation in sparring bouts (P < 0.05) to levels typically observed in steers (P > 0.05). These observations indicate that active immunization against GnRH reduces the incidence of aggressive behavior in male beef cattle and are consistent with our postulate that immunoneutralization of GnRH is an effective alternative to surgical castration in the management of beef cattle. PMID- 12643486 TI - Dietary fat has minimal effects on fatty acid metabolism transcript concentrations in pigs. AB - Young, crossbred pigs were fed either a low-fat, corn-based diet; a high-fat, tallow-based diet with a considerable saturated fatty acid (FA) content; or a high-fat, fish oil-based diet with a considerable polyunsaturated FA content, for 14 d. There were six pigs per dietary group (approximately 4-wk-old with a body weight of 6.16 kg). The plasma and adipose tissue FA composition reflected the composition of the diet to a large extent, but also reflected de novo FA synthesis coupled with chain elongation and desaturation. The liver and skeletal muscle FA composition reflected the diet and endogenous synthesis, but the indications for preferential incorporation or exclusion of specific FA were greater in these tissues than in plasma or adipose tissue. An important transcription factor for adipocyte differentiation and other aspects of lipid metabolism is adipocyte determination and differentiation-dependent factor 1 (ADD1). Liver ADD1 messenger RNA (mRNA) tended to be decreased (P = 0.06) in the fish oil-fed group, as well as in the combined high-fat-fed groups (tallow + fish oil) compared to the low-fat-fed group (P = 0.06). The muscle acyl-CoA oxidase mRNA tended to be increased in the tallow-fed group and decreased in fish oil-fed groups (P = 0.06). The muscle carnitine palmitoyltransferase mRNA tended to be elevated in both fat-fed groups (P = 0.07). None of the adipose tissue mRNA were changed by the diet (P > 0.20). The observations suggest there are major differences between rodents and pigs in modulation of transcripts associated with lipid metabolism by the dietary FA composition or concentration. Also, in porcine adipose tissue, as well as in liver and skeletal muscle, these transcripts are rather refractory to modification by dietary FA. PMID- 12643487 TI - Lipogenic enzyme activities in different adipose depots of Pirenaican and Holstein bulls and heifers taking into account adipocyte size. AB - The effects of sex, genotype, and adipose depot on lipogenic enzyme activity have been investigated in Holstein and Pirenaican bulls and heifers, taking into account differences in adipocyte size. Fifteen Pirenaican bulls and 15 heifers and 15 Holstein bulls and 13 heifers were fattened until slaughter (12 to 13 mo old and 450 to 500 kg of body weight). During the fattening period, animals had ad libitum access to commercial concentrates and straw. The 10th rib was dissected to determine the fat content. Adipocyte size and activities of the following lipogenic enzymes were determined: glycerol 3-phosphate dehydrogenase, fatty acid synthase, nicotinamide adenine dinucleotide phosphate (NADP)-malate dehydrogenase, glucose 6-phosphate dehydrogenase, and NADP-isocitrate dehydrogenase, in the omental, perirenal, subcutaneous, and intermuscular adipose depots, respectively. Because adipocyte mean cell volume varied with sex, breed, and depot, regression analyses of log(e) activity per cell and log(e) cell volume were used to compare activities per unit volume. Sex, breed and depot had no effect (P > 0.05) on the gradients of regressions, which did not differ significantly from 1. Thus, activity per unit volume did not vary with cell size. Consequently, sex, breed, and depot effects on the regression analyses were equivalent to effects on activity per unit volume. Females had greater amounts of fat in the 10th rib (P < 0.001), larger adipocytes (P < 0.001) and, in general, greater (P < 0.05) lipogenic activity per cell, even when adjusted for cell size, than males. These findings suggest that differences in adiposity between sexes are mainly due to females having a greater capacity for lipid synthesis, and hence, hypertrophy, than males. When adjusted for differences in carcass weight, Holsteins had larger adipocytes than Pirenaicans. The abdominal depots, omental and perirenal, had a greater adipocyte size (P < 0.001) and, in general, greater lipogenic enzyme activities per cell (P < 0.05) than the subcutaneous and intermuscular carcass depots. However, when activity per cell was adjusted for cell size, subcutaneous depots had greater fatty acid synthae, glucose 6 phosphate dehydrogenase, and NADP-malate dehydrogenase activities than omental and perirenal, indicating that other factors such as nutrient supply may restrict hypertrophy of carcass adipocytes. PMID- 12643488 TI - Effectiveness of the SmartMV prototype BeefCam System to sort beef carcasses into expected palatability groups. AB - This study was conducted to determine the effectiveness of the SmartMV prototype BeefCam Video Imaging System (prototype BeefCam) for classifying beef carcasses into palatability ("certified" or "not certified" as palatable) groups. Carcasses (n = 769) were selected from four beef-packing plants to represent three USDA quality grade groups (Top Choice, TC; Low Choice, LC; and Select, SE). Following chilling, a prototype BeefCam image of the longissimus muscle was obtained for each carcass. Strip loins were collected from the left side of each carcass and aged for 10 d; Warner-Bratzler shear force (WBSF; n = 769) values and consumer panel ratings (hedonic, end-anchored, 9-point ratings for overall like/dislike; n = 500 carcasses) were obtained for cooked steaks. Using information from the images, two regression models were developed to predict the first principal component of WBSF and consumer panel ratings for sorting carcasses based on expected eating quality. Model I used only prototype BeefCam output, whereas Model II used prototype BeefCam output and a coded value for quality grade group. For both models, carcasses with a predicted value of less than 0.0 were certified as producing palatable beef Additional carcasses (n = 292) were evaluated at a fifth and separate packing plant by prototype BeefCam to validate Models I and II. A strip loin was collected from each carcass and WBSF was measured after 14 d of aging. The percentages of validation carcasses that generated tough (WBSF > or = 4.5 kg) steaks were 6.5,5.8,10.7, and 7.9% for, TC, LC, SE, and all carcasses, respectively. Use of Model I certified 51.9, 47.6, 43.8, and 47.3% of TC, LC, SE, and all carcasses, respectively. Of the carcasses certified by use of Model I, 0.0,0.0, 4.1, and 1.4% of TC, LC, SE, and all carcasses, respectively, generated tough steaks. Use of Model II certified 59.7, 47.6, 25.0, and 42.1% of TC, LC, SE, and all carcasses, respectively. Of the carcasses certified by use of Model II, 2.2, 0.0, 3.6, and 1.6% of TC, LC, SE, and all carcasses, respectively, generated tough steaks. For both models, the frequency of carcasses that produced tough steaks in the certified group was lower (P < 0.05) for all validation carcasses sampled compared with that of the original carcass population. Based on the decrease in the frequency of carcasses that produced tough steaks, further development of a commercial BeefCam system is warranted. PMID- 12643489 TI - Growth performance, carcass and pork quality of finisher pigs fed oat-based diets containing different levels of beta-glucans. AB - A study was undertaken to investigate the growth performance and carcass and meat quality of pigs (BW = 52 to 108 kg) fed oat-based (Avena sativa L.) diets containing four levels of mixed-linkage (1 --> 3), (1 --> 4)-beta-D-glucans. One hundred sixty pigs-80 barrows and 80 gilts (average starting BW = 52.7 kg)--were allocated to one of five diets: a wheat-barley-based control diet and four experimental diets. The groats of Marion, a covered oat, and OT789, a hulless oat, were used to formulate four isonitrogenous and isocaloric diets to achieve 4.1,3.3, 2.1, or 1.6% total /beta-glucans (as fed). Growth performance (daily gain and gain to feed ratio) was not affected (P > 0.05) by the different levels of beta-glucans. Carcass yield, although lower (P < or = 0.05) for pigs fed the control diet, was similar (P > 0.05) for pigs fed any of the experimental diets. Cutout yields were also alike (P > 0.05) across the five diets. Beta-glucan content had no effect (P > 0.05) on the longissimus muscle area, or, by and large, on the proportions of commercial cuts; the only exceptions were a commercial picnic from pigs fed the 2.1% diet lower (P < 0.05) relative to all other diets and a lower (P < 0.5) commercial loin from pigs fed diets 4.1 or 3.3% relative to the control diet. Furthermore, the relative proportions oftotal lean, total bone, and total dissectable fat in the four lean cuts (picnic, butt, loin, and ham) were not different (P > 0.05) among the five diets. For pigs fed 4.1% beta-glucans, the proportion of lean in each of the four major cuts was lower (P < 0.05). No differences (P > 0.05) associated with the level of beta-glucans were detected for either the initial or ultimate pH mean values, the subjective assessment of color or structure of the longissimus muscle, or the instrumentally measured color (L value). Similarly, drip loss was not influenced (P > 0.05) by the level of beta-glucans in the diets. Soluble protein did differ (P < 0.05) among the high- to low-beta-glucans diets. No differences (P > 0.05) associated with diets were found for fat hardness and shear values of grilled pork chops. Chemical fat of the longissimus muscle from pigs fed 4.1, 3.3, or 2.1% beta glucans was lower (P < 0.05) compared to pigs fed the control or 1.6% beta glucans diets. In summary, no evidence of detrimental effect of beta-glucans in oat-based diets, particularly at levels below 4%, was detected, lending support for the inclusion of oat into finisher diets. PMID- 12643490 TI - Online prediction of beef tenderness using a computer vision system equipped with a BeefCam module. AB - Four experiments were conducted in two commercial packing plants to evaluate the effectiveness of a commercial online video image analysis (VIA) system (the Computer Vision System equipped with a BeefCam module [CVS BeefCam]) to predict tenderness of beef steaks using online measurements obtained at chain speeds. Longissimus muscle (LM) samples from the rib (Exp. 1, 2, and 4) or strip loin (Exp. 3) were obtained from each carcass and Warner-Bratzler shear force (WBSF) was measured after 14 d of aging. The CVS BeefCam output variable for LM area, adjusted for carcass weight (cm2/kg), was correlated (P < 0.05) with WBSF values in all experiments. The CVS BeefCam lean color measurements, a* and b*, were effective (P < 0.05) in all experiments for segregating carcasses into groups that produced LM steaks differing in WBSF values. Fat color measurements by CVS BeefCam were usually ineffective for segregating carcasses into groups differing in WBSF values; however, in Exp. 4, fat b* identified a group of carcasses that produced tough LM steaks. Quality grade factors accounted for 3, 18, 21, and 0% of the variation in WBSF among steaks in Exp. 1 (n = 399), 2 (n = 195), 3 (n = 304), and 4 (n = 184), respectively, whereas CVS BeefCam output variables accounted for 17, 30, 19, and 6% of the variation in WBSF among steaks in Exp. 1, 2, 3, and 4, respectively. A multiple linear regression equation developed with data from Exp. 2 accurately classified carcasses in Exp. 1 and 4 and thereby may be useful for decreasing the likelihood that a consumer would encounter a tough (WBSF > 4.5 kg) LM steak in a group classified as "tender" by CVS BeefCam compared with an unsorted population. Online measurements of beef carcasses by use of CVS BeefCam were useful for predicting the tenderness of beef LM steaks, and sorting carcasses using these measurements could aid in producing groups of beef carcasses with more uniform LM steak tenderness. PMID- 12643491 TI - Accuracy of predicting weight and percentage of beef carcass retail product using ultrasound and live animal measures. AB - Five hundred thirty-four steers were evaluated over a 2-yr period to develop and validate prediction equations for estimating carcass composition from live animal ultrasound measurements and to compare these equations with those developed from carcass measurements. Within 5 d before slaughter, steers were ultrasonically measured for 12th-rib fat thickness (UFAT), longissimus area (ULMA), rump fat thickness (URPFAT), and body wall thickness (UBDWALL). Carcasses were fabricated to determine weight (KGRPRD) and percentage (PRPRD) of boneless, totally trimmed retail product. Data from steers born in Year 1 (n = 282) were used to develop prediction equations using stepwise regression. Final models using live animal variables included live weight (FWT), UFAT, ULMA, and URPFAT for KGRPRD (R2 = 0.83) and UFAT, URPFAT, ULMA, FWT, and UBDWALL for PRPRD (R2 = 0.67). Equations developed from USDA yield grade variables resulted in R2 values of 0.87 and 0.68 for KGRPRD and PRPRD, respectively. When these equations were applied to steers born in Year 2 (n = 252), correlations between values predicted from live animal models and actual carcass values were 0.92 for KGRPRD, and ranged from 0.73 to 0.76 for PRPRD. Similar correlations were found for equations developed from carcass measures (r = 0.94 for KGRPRD and 0.81 for PRPRD). Both live animal and carcass equations overestimated (P < 0.01) actual KGRPRD and PRPRD. Regression of actual values on predicted values revealed a similar fit for equations developed from live animal and carcass measures. Results indicate that composition prediction equations developed from live animal and ultrasound measurements can be useful to estimate carcass composition. PMID- 12643492 TI - Efficacy of an E. coli phytase expressed in yeast for releasing phytate-bound phosphorus in young chicks and pigs. AB - Four chick trials and one pig trial were conducted to investigate the phosphorus releasing efficacy oftwo commercial phytase enzymes (Natuphos and Ronozyme) and an experimental E. coli phytase enzyme (ECP) when added to corn-soybean meal diets containing no supplemental inorganic P (iP). In the 13- or 14-d chick trials, three or four graded levels of iP (0, 0.05,0.10,0.15%) from KH2PO4 were added to the basal diet to construct standard curves from which bioavailable P release could be calculated for the phytase treatments. In all cases, phytase supplementation levels were based on an assessment of phytase premix activity (i.e., P release from Na phytate at pH 5.5). Linear (P < 0.01) responses in tibia ash and weight gain resulted from iP supplementation in all assays. In the first chick trial, supplementation of 500 phytase units (FTU)/kg of ECP resulted in superior (P < 0.01) weight gain and tibia ash values compared with 500 FTU/kg of Natuphos. Results of the second chick trial revealed P-release values of 0.032 and 0.028% for 500 FTU/kg Natuphos and Ronozyme, respectively, and these were lower (P < 0.01) than the 0.125% P-release value for 500 FTU/kg of ECP. Tibia ash responded quadratically (P < 0.05) in response to graded levels of ECP up to 1,500 FTU/kg in the third chick trial. Combining Natuphos with either Ronozyme or ECP in Chick Trial 4 revealed no synergism between phytases with different initiation sites of P removal. The pig trial involved 10 individually fed weanling pigs per diet, and and phytase enzymes were supplemented to provide 400 FTU/kg in diets containing 0.60% Ca. Based on the linear regression of fibula ash on supplemental iP intake (r2 = 0.87), P-release values were 0.081% for Natuphos, 0.043% for Ronozyme, and 0.108% for ECP. These trials revealed an advantage of the E. coli phytase over the commercial phytases in young chicks. PMID- 12643493 TI - Variability in mixing efficiency and laboratory analyses of a common diet mixed at 25 experiment stations. AB - An experiment involving 25 experiment stations in the North Central and Southern regions (NCR-42 and S-288, respectively) was conducted to assess the degree of uniformity of diet mixing among stations and to assess the variability among station laboratories in chemical analysis of mixed diets. A fortified corn soybean meal diet was mixed at each station using a common diet formula (except for vitamin and trace-mineral additions). The diet was calculated to contain 14% crude protein (CP), 0.65% Ca, 0.50% P, and 125 ppm Zn (based on 100 ppm added Zn). After mixing, samples were collected from the initial 5% of feed discharged from the mixer, after 25, 50, and 75% was discharged, and from the final 5% of discharged feed. The five samples were sent to the University of Kentucky, finely ground, and divided into subsamples. Each set of five subsamples from each station was distributed to three randomly selected stations for analysis of CP, Ca, P, and Zn (i.e., each station analyzed five diet sub-samples from three other stations). In addition, two commercial and two station laboratories analyzed composites of the five subsamples from each of the 25 mixed diets. Based on the laboratories that analyzed all diets, means were 13.5, 0.65, and 0.52%, and 115 ppm for CP, Ca, P, and Zn, respectively. Ranges of 11.8 to 14.6% CP, 0.52 to 0.85% Ca, 0.47 to 0.58% P, and 71 to 182 ppm of Zn were found among the 25 diet mixes. The coefficients of variation among the 25 diet samples for CP, Ca, P, and Zn were 4.3, 9.3, 4.1, and 17.4%, and among the 25 laboratories were 3.6, 12.5, 10.7, and 11.1%, respectively. Overall analyses of the five sub samples were, respectively, CP: 13.4, 13.6, 13.4, 13.5, and 13.4% (P < 0.06); Ca: 0.66, 0.67, 0.67, 0.66, and 0.67%; P: 0.50,0.51,0.51,0.50, and 0.50%; and Zn: 115, 116, 112, 113, and 120 ppm (P < 0.001). Diets were not uniformly mixed at all stations (station x sample No. was P < 0.08 for Ca and P < 0.01 for CP, P, and Zn). Among stations, the range of the five samples, expressed as a percentage of the mean and averaged for CP, Ca, P, and Zn, varied from +/- 1.1% (i.e., 98.9 to 101.0%) to +/- 12.9% (84.6 to 110.4%), with an overall average of +/- 5.2%. Neither type nor volume of mixers was related to mixing uniformity. The results suggest that uniformity of diet mixes varies among experiment stations, that some stations miss their targeted levels of nutrients (especially Zn), and that the variability among experiment station laboratories in analysis of dietary Ca, P, and Zn in mixed diets is quite large. PMID- 12643494 TI - Effects of adding fiber sources to reduced-crude protein, amino acid-supplemented diets on nitrogen excretion, growth performance, and carcass traits of finishing pigs. AB - Two experiments were conducted to evaluate the effects of adding fiber sources to reduced-crude protein (CP), amino acid-supplemented diets on N excretion, growth performance, and carcass traits of growing-finishing pigs. In Exp. 1, six sets of four littermate barrows (initial weight = 36.3 kg) were allotted randomly to four dietary treatments to determine N balance and slurry composition. Dietary treatments were: 1) fortified corn-soybean meal, control, 2) as fortified corn soybean meal with CP lowered by 4 percentage units and supplemented with lysine, threonine, methionine, tryptophan, isoleucine, and valine (LPAA), 3) same as Diet 2 plus 10% soybean hulls, and 4) same as Diet 2 with 10% dried beet pulp. Nitrogen intake, absorption, and retention (g/d) were reduced (P < 0.04) in pigs fed the low- protein diets, but they were not affected (P > 0.10) by addition of fiber sources to the LPAA diet. However, N absorption, as a percentage of intake, was not affected (P > 0.10) by dietary treatment. Nitrogen retention, expressed as a percentage of N intake, was increased (P < 0.02) in pigs fed the low-protein diets, but it was not affected by fiber addition to the LPAA diet. Urinary and total N excretion was reduced (P < 0.01) by 50 and 40%, respectively, in pigs fed the low- protein diets, but it was not affected (P > 0.10) by fiber addition. However, fiber addition to the LPAA diet tended to result in a greater proportion of N excreted in the feces than in the urine. Slurry pH, ammonium N content, and urinary urea N excretion were reduced (P < 0.10) in pigs fed LPAA, and a further reduction (P < 0.06) in slurry ammonium N content and urinary urea N was observed with fiber addition. Also, fiber addition to the LPAA diet increased (P < 0.02) slurry VFA concentrations. In Exp. 2, 72 pigs were blocked by body weight and sex and allotted randomly to three dietary treatments that were similar to those in Exp. 1, with a corn-soybean meal control diet, LPAA diet, and a LPAA diet with 10% soybean hulls. Pigs were fed the diets from 28.6 to 115 kg, and all pigs were killed for collection of carcass data. Growth performance and most carcass traits were not affected (P > 0.10) by dietary treatment. These data suggest that reducing CP with amino acid supplementation markedly decreased N excretion without influencing growth performance. Fiber addition to a LPAA diet had little effect on overall N balance or growth performance, but tended to further reduce slurry ammonium N concentration and increase volatile fatty acid concentrations. PMID- 12643495 TI - Relationships of testicular iron and ferritin concentrations with testicular weight and sperm production in boars. AB - The inverse relationship of testicular size and circulating follicle-stimulating hormone (FSH) concentrations has been documented, and accompanying this relationship is the change in color of the parenchymal tissue of the testes. Large testes (300 to 400 g) are pink to light red and small testes (100 g) are dark maroon with color gradations for weights in between. It was hypothesized that this color most likely represented an iron protein. Chromatographic analysis of testicular tissue indicated that the Fe was associated primarily with ferritin, and immunohistochemistry showed that Leydig cells were the primary location of ferritin storage within the testes. Concentrations of Fe and ferritin were higher in small testes and decreased as testes weight increased (P < 0.05). As testicular Fe concentrations increased, daily sperm production (DSP) and total DSP declined (P < 0.05). Genotyping six generations of Meishan x White composite boars (n = 288) for a quantitative trait locus that is indicative of elevated FSH and small testes in boars indicated that the Meishan genotype had elevated testicular iron concentrations and darker color in conjunction with reduced total DSP (P < 0.01). It is not thought the elevated iron concentrations affect testicular weights but are probably a result of elevated FSH and FSH inducement of Fe transport. The storage of Fe in Leydig cells may provide a reservoir of Fe for easy access by Sertoli and germ cells, but still provide a degree of protection to germ cells from ionic iron. PMID- 12643496 TI - Dehydration in stressed ruminants may be the result of a cortisol-induced diuresis. AB - The effect on water and electrolyte balance of stress, simulated by intravenous infusion of cortisol, was studied using 24 18-mo-old Merino wethers (37.0 +/- 0.94 kg mean body weight [BW]) over 72 h. The sheep were allocated to one of four groups: 1) no water/no cortisol (n = 6); 2) water/no cortisol (n = 4); 3) no water/cortisol (n = 6); and 4) water/cortisol (n = 4). Animals allocated to the two cortisol groups were given 0.1 mg x kg BW(-1) x h(-1) of hydrocortisone suspended in isotonic saline to simulate stress for the duration of the experiment. Total body water, plasma cortisol, osmolality and electrolytes, and urine electrolytes were determined at 24-h intervals for 72 h. In the presence of cortisol, total body water was maintained in the face of a water deprivation insult for 72 h. Water deprivation alone did not induce elevated plasma concentrations of cortisol, in spite of a 13% loss of total body water between 48 and 72 h. Infusion of cortisol was found to increase urine output (P = 0.003) and decrease total urinary sodium output (P = 0.032), but had no effect on plasma electrolyte levels or water intake. Water deprivation was found to increase plasma sodium concentrations (P = 0.037). These results indicate that sheep given cortisol to simulate stress suffer from a loss of body water in excess of that associated with a loss of electrolytes, and support the hypothesis that elevated physiological concentrations of cortisol induce a diuresis in ruminants that contributes to dehydration. PMID- 12643497 TI - Simplified in situ method for estimating ruminal dry matter and protein degradability of concentrates. AB - In this study, dry matter and crude protein in situ degradation data from different concentrate feeds were used to test the accuracy of effective degradability (ED) measures when using reduced ruminal incubation times compared with models based on seven or eight incubation times. The ED was estimated both with and without correction for nylon bag particle loss. The crude protein ED corrected for particle loss of the calibration data set was widely distributed in a range from 16 to 90% with an overall mean value of 60.4%, and the dry matter ED was distributed in the range from 22.7 to 80.7%, with a mean value of 56.9%. The simplified method was developed based on bilinear regression models where all combinations of one to three disappearance values were tested to find the optimal time point combinations to estimate ED. Bilinear regression models based on two and three ruminal incubation times gave similar estimates to a standard in situ method over a wide range of passage rates both for the data set used to parameterize the models and the independent data set used to evaluate the models. Using two incubation times, the bilinear model based on 4 and 24 h gave the most accurate estimates, and the models based on 2, 8, and 24 h for uncorrected data and 4, 8, and 24 h for corrected data were most accurate of the three time points bilinear models. The number of nylon bags used by these models was reduced by 58 to 78% compared with the standard in situ method, and the total incubation time needed was substantially reduced. PMID- 12643498 TI - Changes in single unit activity in the lateral hypothalamic area of goats during feeding. AB - In order to clarify the location of feeding centers in the ruminant brain, this study used a single-unit activity (SUA) recording electrode to investigate the existence of appetite-regulating neurons in the lateral hypothalamic area (LHA) in goats. Seven male Japanese Saanen goats were used in the experiment. The animals were fed twice daily, once in the morning (1000 to 1200) with 1.5 kg of roughly crushed alfalfa hay cubes, and once in the afternoon (1600 to 1800) with 200 g of commercial ground concentrate feed. The animals were allowed free access to drinking water. In this study, the animals were surgically operated on to position the recording electrode in the LHA. Recordings of SUA were carried out continuously over a 2.25-h period beginning 15 min prior to the commencement of morning feeding. The eating rates of crushed alfalfa hay cubes were highest 10 min after feeding commencement, but decreased sharply by the time 40 min had elapsed. The cumulative feed intake after the completion of the 2-h feeding period was 1164 +/- 38 g. The cumulative water intake upon the conclusion of the 2-h feeding period was 2422 +/- 107 mL. This study recorded 31 units, of which five showed a response to feeding and altered their firing rates. In response to a sharp increase in eating rates, all five units increased their firing rates to a level higher than that of prefeeding (P < 0.05). As the animals reached a level of satiety (eating rates declined to very low levels), firing of units I and II stopped completely, while the firing rates of units III, IV, and V decreased. Examination of a serial histological section confirmed that the five units in which changes in firing rates with feeding were observed were all located in the dorsolateral hypothalamic area close to the fornix. The LHA neurons recorded in this experiment characteristically showed neuronal activity increases at high levels of feeding, but decreases at low levels. The results suggest that there are cells located in the LHA of goats that are active in the physiological regulation of hay (dry forage) intake. PMID- 12643499 TI - Diet and procedures used to detach particle-associated microbes from ruminal digesta influence chemical composition of microbes and estimation of microbial growth in Rusitec fermenters. AB - Four different detachment methods were evaluated for their ability to remove particle-associated microorganisms (PAM) from ruminal digesta in semicontinuous fermenters fed two diets differing in their forage:concentrate ratio (80:20 [C20] and 20:80 [C80]). In the methylcellulose method, ruminal digesta was incubated at 38 degrees C for 15 min with saline solution containing 0.1% methylcellulose before being stored at 4 degrees C for 24 h. In the other procedures, samples were incubated with 0.1% methylcellulose before storage for 24 h at 4 degrees C in different solutions (pH = 2): 1) saline solution with 0.1% Tween 80; 2) saline solution with 0.1% Tween 80 and 1% tertiary butanol; and 3) saline solution with 0.1% Tween 80, 1% methanol, and 1% tertiary butanol. Common to all treatments was subsequent homogenization, followed by filtration and resuspension of the residue five times in the treatment solutions. Microbial removal was estimated indirectly by measuring removal of 15N. There were no differences (P > 0.05) among detachment procedures, neither in the detaching efficiency (mean values of 79.7 and 88.1% for C20 and C80 diets, respectively) nor in the total recovery of PAM (54.9 and 34.9% for C20 and C80, respectively). There were no differences (P > 0.05) among PAM pellets obtained by the different detachment procedures in their N content, purine bases (PB) concentration, or PB:N ratio. For the C80 diet, 15N enrichment was greater (P < 0.05) in PAM pellets obtained with methylcellulose than in those obtained by the other methods. However, there were no differences (P > 0.05) due to the detachment procedure in the values of daily microbial growth estimated using as reference the different PAM pellets. The PAM pellets for diet C20 presented greater (P < 0.01) N content and lower (P < 0.01) PB concentration than those for diet C80 (mean values of 74.3 vs 49.1 mg of N/g of dry matter, and 22.8 vs 26.0 micromol PB/mg of dry matter, respectively). Daily microbial growth was greater (P < 0.05) for the C80 diet than for the C20 diet (121 vs 114 mg of microbial N, respectively). Results suggest that the treatment of ruminal digesta with a saline solution with 0.1% methylcellulose at 38 degrees C for 15 min combined with homogenizing and chilling at 4 degrees C for 24 h removed a major proportion of PAM, although further research is needed to decrease microbial losses during the isolation process. PMID- 12643500 TI - Nitrogen metabolism and recycling in Holstein heifers. AB - To study the effect of dietary N level on urea kinetics and recycling, four Holstein heifers (267 +/- 3.6 kg) were used in a Youden square design. Isocaloric diets with a N content of 1.44, 1.89, 2.50, 2.97, and 3.40% were fed at approximately 1.8 times maintenance intake. Increasing the N content of the diet increased urinary N excretion (P < 0.001) and N balance (P < 0.01), but did not affect the fecal N excretion (P = 0.21). Increasing the level of dietary N, increased urea production (P < 0.001) and excretion (P < 0.001), but no effect (P = 0.24) could be detected in the amount of N recycled to the gut. Urea recycled with the saliva, however, increased (P < 0.001) both in absolute and relative terms, with increasing dietary N. No difference could be detected on the amount of recycled N that was used for anabolism or returned to the ornithine cycle, but less (P = 0.001) N originating from urea was excreted in feces as dietary N increased. Ruminal ammonia concentration increased (P < 0.001) with increasing N intake, but total tract neutral detergent fiber digestibility was depressed only on the lowest N intake diet. No difference (P = 0.30) was detected in ruminal microbial yield among diets, but more (P < 0.003) N was derived from blood urea at low N intakes, and the efficiency of use of the recycled N decreased (P < 0.001) with increasing levels of dietary N. Adaptive changes to low-N diets were a decrease (P < 0.003) in the renal clearance of urea and an increase (P < 0.001) in the gastrointestinal clearance of urea. Urea transporters were present in the rumen wall of the heifers and differentially expressed depending on dietary N content, but their role in the transfer of urea into the rumen remains uncertain. Different mechanisms of N salvage and recycling were involved when animals were fed low-N diets that ensured a supply of endogenous N to the gastrointestinal tract and, due to the reduced contribution of dietary N, an increased efficiency of the N recycled was observed. PMID- 12643501 TI - Effect of feeding the ionophores monensin and laidlomycin propionate and the antimicrobial bambermycin to sheep experimentally infected with E. coli O157:H7 and Salmonella typhimurium. AB - Escherichia coli O157:H7 and Salmonella are widely recognized as important agents of foodborne disease with worldwide distribution. The use of ionophores in feeding growing ruminants is widespread in the United States and has attracted recent interest due to the apparent temporal relationship between initial ionophore use and the increase in human E. coli O157:H7 cases. Two experiments were conducted to evaluate the effects of short-term feeding of ionophores on fecal shedding, intestinal concentrations, and antimicrobial susceptibility of E. coli O157:H7 and S. typhimurium in growing lambs. Sixteen lambs were used in each experiment, four lambs per treatment group: monensin, laidlomycin propionate, bambermycin, and a control treatment. Lambs were fed a grain and hay (50:50) diet with their respective ionophore for 12 d before experimental inoculation with E. coli O157:H7 or S. typhimurium. Animals were maintained on their respective diets an additional 12 d, and fecal shedding of inoculated pathogens was monitored daily. Lambs were killed and tissues and contents were sampled from the rumen, cecum, and rectum. No differences (P > 0.05) in fecal shedding of Salmonella or E. coli O157:H7 were observed due to treatment. Occurrence of Salmonella or E. coli in luminal contents and tissue samples from the rumen, cecum, and rectum did not differ (P > 0.05) among treatments. Feeding monensin decreased (P < 0.05) the incidence of scours in sheep infected with Salmonella compared with the other treatments. No differences in antimicrobial susceptibility were found in any of Salmonella or E. coli O157:H7 isolates. Results from these studies indicate that short-term ionophore feeding had very limited effects on E. coli and Salmonella shedding or on antimicrobial susceptibility in experimentally infected lambs. PMID- 12643502 TI - Effect of carrier materials on the resistance of spores of Bacillus stearothermophilus to gaseous hydrogen peroxide. AB - The testing of the H2O2 decontamination process using spores of Bacillus stearothermophilus has gained widespread acceptance. Usually, commercially available Biological Indicators (BIs) with a specified resistance to H2O2 are challenged to qualify the process. The question arises whether the resistance of test spores is dependant on the type of carrier material and whether the resistance is representative for the system under test. The objective of the study is to quantify the effect of different carrier materials on the resistance of spores of Bacillus stearothermophilus to H2O2. Materials from which isolators were built, as well as those used in disposables during daily work were investigated. These materials were inoculated with 106 spores of Bacillus stearothermophilus (ATCC 7953). The spore resistance was tested to a well defined H2O2 decontamination cycle by determining the D-value using the "Fractional Negative" method. This paper reports on the effect of different carrier materials to the resistance of the test organism against H2O2. Various materials have significantly increased resistance of the spores and should be avoided in isolator systems. If commercially available BIs are used for process qualification, the resistance of the BI used, the fluctuation in resistance caused by isolator materials, the required log reduction, and at least the bioload of isolator surfaces need to be known. PMID- 12643503 TI - An automated sequential injection analysis system for the determination of trace endotoxin levels in water. AB - A new automated instrument based on the Limulus amebocyte lysate (LAL) chromogenic substrate kinetic assay for the determination of bacterial endotoxins is reported. A computer controlled syringe pump-multiport valve combination was used to aspirate the sample and other reagents into a holding coil. The syringe was always filled with air; liquid did not enter the syringe. The valve could address up to four individual radial paths, fully referenced optical fiber LED based absorbance detectors that were housed collectively in a single, small (20 x 20 x 30 mm) metal block with a heater and temperature sensor, and maintained at 37 degrees C. Assay mixtures containing sample or standards were delivered to the individual detector cells for the simultaneous collection of the time based absorbance data. The automated system determined endotoxins with good accuracy and reproducibility in the range of 0.005-0.5 endotoxin units (EU)/mL (r2 > or = 0.99). Based on three times the standard deviation of the blank and the slope of the calibration curve, the lower limit of detection was < or = 0.003 EU/mL. The variability of the assay method is less than 5% (n = 10). Analysis time required for a 0.005 EU/mL standard was <100 min. Appropriately diluted tap water samples were simultaneously analyzed by the present system and a manually loaded commercial microplate reader based instrument; the data were statistically indistinguishable at the 95% confidence level. PMID- 12643504 TI - Qualification of a rapid readout biological indicator with moist heat sterilization. AB - Biological indicators are recognized as an important component in the validation and routine monitoring of moist heat (steam) sterilization processes. Due to the need to allow for the recovery and outgrowth of test organisms that may have been sub-lethally injured, between 2-5 days of incubation are typically required before the outcome of sterilization processing can be reliably interpreted. Rapid readout biological indicators that incorporate the response of a heat resistant enzyme provide a means for assessing the efficacy of moist heat sterilization within hours of processing. This study describes the qualification of the 3M Attest 1292 Rapid Readout Biological Indicator with moist heat sterilization according to the procedures described in the PDA Technical Report No. 33, "Evaluation, Validation and Implementation of New Microbiological Testing Methods". PMID- 12643505 TI - Use of the hazard analysis and critical control points (HACCP) risk assessment on a medical device for parenteral application. AB - In order to guarantee the consistently high quality of medical products for human use, it is absolutely necessary that flawless hygiene conditions are maintained by the strict observance of hygiene rules. With the growing understanding of the impact of process conditions on the quality of the resulting product, process controls (surveillance) have gained increasing importance to complete the quality profile traditionally defined by post-process product testing. Today, process controls have become an important GMP requirement for the pharmaceutical industry. However, before quality process controls can be introduced, the manufacturing process has to be analyzed, with the focus on its critical quality influencing steps. The HACCP (Hazard Analysis and Critical Control Points) method is well recognized as a useful tool in the pharmaceutical industry. This risk analysis, following the guidelines of the HACCP method and the monitoring of critical steps during the manufacturing process was applied to the manufacture of methyl methacrylate solution used for bone cement and led to the establishment of a preventative monitoring system and constitutes an effective concept for quality assurance of hygiene and all other parameters influencing the quality of the product. PMID- 12643507 TI - Selecting a training documentation/recordkeeping system in a pharmaceutical manufacturing environment. PMID- 12643506 TI - Development of an advanced high speed aseptic filling system. AB - A new pharmaceutical aseptic vial filling system has been developed based on isolator technology that is also well suited to use in conventional clean rooms. The results of experiments designed to test the microbiological quality of the environment provided by the new filling system in conjunction with an isolator demonstrate that the isolator enclosed filler can be readily decontaminated and that it can maintain an aseptic environment for at least one month under worse case conditions. PMID- 12643508 TI - Bill & Melinda Gates Foundation announces dollar 200 million grant to accelerate research on biggest problems in global health. PMID- 12643509 TI - Elias Zerhouni offers solutions for "broken" clinical research enterprise. PMID- 12643510 TI - Translating clinical research into practice: practice-based research networks--a promising solution. AB - Greater effort is needed to translate research discoveries into clinical practice and to ensure that proven treatments are routinely implemented by physicians. Practice-based research networks are an undervalued resource in the endeavor to bring research results to bear in patient care. These networks are groups of practicing health care providers who have organized to address research questions of primary importance to their patients. Existing practice-based research networks have addressed a wide range of issues, including improvement of preventive service delivery, reliability of diagnostic measures, practice variation, disease prevention, and systems of care coordination. Several deficiencies in the current system prevent practice-based research networks from reaching their full potential as mechanisms of translating clinic research into practice. For practice-based research networks to flourish, they need increased funding for practice-based research networks; information systems to facilitate research by practicing physicians; training programs for health care providers; and communication networks between providers, clinical researchers, professional societies, and academic health centers. PMID- 12643511 TI - Improving the US health care system: action plan to enhance efficiency, reduce errors, and improve quality. AB - The decisions made by stakeholders in the nation's health care system that affect the quality of care experienced by patients are too often made without the benefit of scientific evidence. A multidisciplinary set of investigators conducting health services research have traditionally filled this gap between research findings and clinical decision making, but several barriers are hindering this work. This article offers several recommendations-restructuring organizations, ensuring funding, developing infrastructure, strengthening the community of researchers, and forging new links among stakeholders-to promote high-quality information for health decision makers. PMID- 12643512 TI - Mitogenic effects of Brazilian arthropod venom on isolated islet beta cells: in vitro morphologic ultrastructural and functional studies. AB - BACKGROUND: One of the major pitfalls associated with use of isolated adult islets of Langerhans' cells is their minimal mitotic capacity. Consequently, maintenance of a steady viable islet cell mass is very difficult. To explore how to enhance beta-cell mitogenesis, we have examined the effects of venom fractions extracted from a Brazilian scorpion on morphologic and functional beta-cell patterns. The venom was previously known to induce nesidioblastosis-like effects with chronic hypoglycemia and pancreatitis in animal models. METHODS: Venom fractions purified from Tityus bahiensis were incubated with batches of isolated rat islets, while a morphologic examination, glucose-stimulated insulin release, insulin content, and insulin messenger ribonucleic acid (mRNA) were carried out early during incubation. On fixation and double fluorescence immunolabeling (rhodamine for anti-insulin monoclonal antibodies; fluorescein for anti-5 bromodeoxyuridine), the preparations were imaged by confocal laser microscopy (CLM) for morphometric quantification of the mitoses. Insulin recovery and mRNA were also assessed at 21 days of culture. RESULTS: Under CLM examination, the beta-cell mitotic rate significantly rose from 1 to 12.8% for the venom-exposed islets. At day 7, insulin release and content were significantly lower for the venom-exposed than the control islets. However, at day 21 of culture, insulin release in response to static incubation with glucose and insulin mRNA from the venom-exposed islets was higher than controls (p < .05). CONCLUSIONS: Incubation with the scorpion venom induced a rapid and significant increase in the beta-cell proliferation not associated with a short-term increase in insulin secretion. The latter fully resumed and overcame controls later in culture, possibly after completion of the beta-cell expansion process. PMID- 12643513 TI - Cardiac mitochondrial calcium loading capacity is severely affected after chronic cholestasis in Wistar rats. AB - BACKGROUND: Cardiovascular changes correlated with some forms of hepatic disease are being reported in the literature. OBJECTIVES: The aim of this work was to characterize cardiac mitochondrial bioenergetics and calcium buffering capacity in Wistar rats injected with six weekly doses of alpha-naphthylisothiocyanate (ANIT), a compound known to induce cholestasis in animal models. METHODS: Isolated heart mitochondria were obtained from both injected and control animals and bioenergetic parameters were measured, as well as the capacity to buffer externally added calcium and the mitochondrial content of reduced protein thiol groups. Blood biochemistry analyses were obtained at the initial and end points of treatment. The in vitro ANIT effect on isolated heart mitochondria was also studied. RESULTS AND DISCUSSION: Our results showed that the respiratory control ratio was the only parameter affected in injected animals (p < .05, n = 5). Nevertheless, heart mitochondria from injected animals showed an inability to accumulate added calcium owing to an increased susceptibility to the calcium dependent mitochondrial permeability transition (p < .0001, n = 5). The effects were still present 1 week after ending ANIT administration, when serum markers for liver injury and hyperbilirubinemia were already abated (although in the presence of bile duct proliferation). To our knowledge, this is the first time that cardiac mitochondrial calcium homeostasis and mitochondrial respiratory ratio are seen affected during ANIT-induced cholestasis, prevailing even in the absence of hepatic damage serum markers. PMID- 12643514 TI - Silica, hyaluronate, and alveolar macrophage functional differentiation. AB - BACKGROUND: Silicosis is mediated by macrophages, their soluble mediators, and extracellular matrix molecules. In this study, we investigated the effects of silica and/or hyaluronate (HA) on several alveolar macrophage responses. METHODS: We evaluated glycosaminoglycan (GAG) production by radiolabeled precursors, nitric oxide (NO) release by its oxidation product, phagocytic activity by Candida albicans internalization, and the secretion of two fibrogenic cytokines, tumor necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta, by specific assays. RESULTS: Silica significantly reduced GAG secretion, particularly HA secretion. Alone, it decreased Candida uptake; associated with HA, it enhanced the reduction. Silica and Candida reduced NO release, which was not significantly affected when silica- or Candida-exposed cells were also treated with HA. TNF-alpha and TGF-beta activities were stimulated by silica but reduced by HA. CONCLUSIONS: The results suggest that silica and HA modify alveolar macrophage functional differentiation. Silica- and HA-induced modifications of the microenvironment could determine whether the response proceeds toward healing and repair or toward lung chronic pathology. PMID- 12643515 TI - Abnormalities of serum antielastin antibodies in connective tissue diseases. AB - BACKGROUND: Antibodies (Abs) to alpha-elastin (elastin breakdown product) and tropoelastin (elastin precursor) are found in the serum of all human subjects and correlate with their respective serum peptide levels; however, peptide levels vary with age and some disease states. This study was undertaken to determine if serum elastin Abs, peptides, and elastin metabolism were altered in autoimmune diseases by detecting a changing ratio of serum anti-alpha:tropoelastin Ab levels. METHODS: Serum from patients with a variety of connective tissue diseases, including 28 with systemic lupus erythematosus (SLE), 24 with scleroderma, 18 with rheumatoid arthritis (RA), 10 with polymyositis, and 39 with vasculitis, was compared with serum from 19 age-matched healthy subjects for levels of antitropoelastin and anti-alpha-elastin Abs. RESULTS: We found an increase in IgG anti-alpha-elastin and a decrease in antitropoelastin Abs in the sera of patients with scleroderma (p < .02 and .00005) and SLE (p < .006 and .011). There was also a marked increase in anti-alpha-elastin Abs in patients with polyarteritis nodosa (p < .0005) and decreases in antitropoelastin Abs in patients with RA (p < .05), polymyositis (p < .01), and a variety of other vasculidities (p < .0003). CONCLUSIONS: Abnormal variations in elastin metabolism may be detected in several connective tissue diseases by measuring ratios of alpha- and tropoelastin IgG Abs as markers of elastin degradation and synthesis. PMID- 12643516 TI - Motherhood at or beyond the edge of reproductive age. AB - Motherhood at or beyond the edge of reproductive age is a new aspect of what clinicians previously referred to as pregnancy in the "older gravida." With the implementation of infertility treatment, the boundary between "old" and "young" no longer exists. Currently, three types of old mothers are seen in the physician's office: those who accidentally conceive, those who still have fertilizable oocytes for assisted reproduction, and those conceiving with assisted reproduction after egg donation. In the United States alone, the 2001 rates for births to women aged 35-39, 40-44, and 45-49 years rose 30, 47, and 190% compared with 1990. Specifically, there were about 5,000 births to women > or = 45 years. Generally, the majority of the published studies have been unanimous about the special, and perhaps the super-cautious attitude required for the older mother, an approach that translates to higher rates of peripartum interventions. This is despite the fact that although some complications may occur more frequently in older mothers as a result of accumulated prior diseases, there is no direct evidence that older age per se complicates either gestation or parturition. In contrast, the risks for the fetus-neonate depend on the source of the oocytes and relate to the mode of conception. Also, older age has an advantage of better perinatal outcome of twins and triplets. Despite many unanswered questions, clinicians should exercise an unbiased approach and be prepared for the challenges and potential complications related to patients who are older than those seen in the past. The "unnatural" connotation of a pregnancy at an advanced maternal age should be accepted as a consequence of the combined effect of social changes and medical progress and not as a counter-evolutionary phenomenon. PMID- 12643517 TI - Oocyte quality and IVF outcome after coasting to prevent ovarian hyperstimulation syndrome. AB - OBJECTIVE: To evaluate, in a large cohort of patients, oocyte quality and IVF outcome after coasting used to prevent ovarian hyperstimulation. SETTING: Retrospective study. PATIENTS: IVF cycles which had reached estradiol serum levels of at least 4,000 pg/mL without being coasted (control group, n = 208), or where coasting was applied (coasted group, n = 157). METHOD: IVF data of coasted cycles were compared with the control group. Within the group of coasted cycles, we also analyzed whether indirect parameters related to coasting had an effect on IVF results. RESULTS: Coasted patients showed higher maximum estradiol levels and greater numbers of large follicles than the control group, but lower oocyte recovery rates. There were no other significant differences between the two groups of patients. Within the group of coasted patients, no significant relation was found between the number of days of coasting, the estradiol level on the day of hCG, or the fall in estradiol and the outcome, whether measured in terms of oocyte quality, pregnancy rate, or incidence of ovarian hyperstimulation. CONCLUSION: Coasting seems to be associated with a reduced oocyte collection rate, especially when the coasting period is prolonged. However, this does not result in reduced oocyte quality. The length of the coasting period and degree of estradiol decrease do not seem to alter the results in terms of pregnancy rates. PMID- 12643518 TI - Can we decrease breakthrough bleeding in patients with endometriosis on norethindrone acetate? AB - OBJECTIVE: To compare the breakthrough bleeding in endometriosis patients treated with Lupron-Depot alone, norethindrone acetate following Lupron-Depot, and norethindrone acetate alone. PATIENTS AND METHODS: 71 women with symptomatic surgically diagnosed endometriosis were retrospectively evaluated for this study. 28 women were treated with 6 doses of 3.75 mg Lupron-Depot every 4 weeks (Group I). 15 women were treated first with Lupron-Depot 3.75 mg every 4 weeks for 3 to 6 doses, followed by 5 mg norethindrone acetate (Group II). 28 patients were treated for 6 months with 5 mg per day norethindrone acetate alone (Group III). Breakthrough bleeding during treatment was scored mild (some spotting), moderate (lighter than patient's normal menstruation), or severe (as much as patient's normal menstruation or heavier). Multiple comparisons were done by ANOVA (SPSS) among three groups. The age of patients was not significantly different between groups 134.9-36.8 years). BMI of the three groups was significantly different 126.6 +/- 5.8, 27.4 +/- 6.4, 23.6 +/- 4.5, respectively). RESULTS: Breakthrough bleeding was reported by 14% of Group I, 20% of Group II, and 68% of Group III. CONCLUSION: Endometriosis patients who were treated with norethindrone acetate following Lupron-Depot had significantly less breakthrough bleeding than those given norethindrone acetate alone, and the incidence was comparable to Lupron Depot alone. PMID- 12643520 TI - The challenges ahead. PMID- 12643519 TI - Triplet pregnancies and their aftermaths II. Risks of older mothers and plurality. AB - There is no literature that accurately reports the risks inherent to the woman over age 40 who carries a triplet pregnancy. Extrapolating the singleton literature does not provide solid evidence, since risks are described variously by different authors and may relate to maternal age or parity as independent factors, which affect both maternal and fetal outcome data. On the other hand, recent population-based surveys strongly suggest that plurality is a more important risk factor than maternal age, for it affects both maternal and fetal outcomes. PMID- 12643521 TI - Personal commentary on proteomics. PMID- 12643522 TI - DTASelect and Contrast: tools for assembling and comparing protein identifications from shotgun proteomics. AB - The components of complex peptide mixtures can be separated by liquid chromatography, fragmented by tandem mass spectrometry, and identified by the SEQUEST algorithm. Inferring a mixture's source proteins requires that the identified peptides be reassociated. This process becomes more challenging as the number of peptides increases. DTASelect, a new software package, assembles SEQUEST identifications and highlights the most significant matches. The accompanying Contrast tool compares DTASelect results from multiple experiments. The two programs improve the speed and precision of proteomic data analysis. PMID- 12643524 TI - Design and synthesis of class-selective activity probes for protein tyrosine phosphatases. AB - Two mechanism-based activity probes, adopting a cassette-like design, for protein tyrosine phosphatases (PTPs) were synthesized. Both probes carry a phosphate group that serves as the recognition head for the target PTPs but differ in their reporter groups; probe LCL-1 uses a dansyl fluorophore, while LCL-2 has a biotin reporter group. LCL-1 and LCL-2 are specifically activated by phosphatase, leading to its covalent labeling, as exemplified with PTP-1B. However, they show no activation with other classes of hydrolases, including trypsin and beta galactosidase. LCL-1 and LCL-2 thus represent the first example of class selective probes for phosphatases. PMID- 12643523 TI - Proteolytic 18O labeling for comparative proteomics: evaluation of endoprotease Glu-C as the catalytic agent. AB - Recently, proteolytic 18O labeling has been demonstrated as a promising strategy for comparative proteomic studies (Yao, X.; Freas, A.; Ramirez, J.; Demirev, P. A.; Fenselau, C. Anal. Chem. 2001, 73, 2836-42). In this approach, protein mixtures are digested in parallel in H216O and H218O and the ratios of isotopically distinct peptide products are measured by mass spectrometry. In the initial report from this laboratory, trypsin was shown to catalyze incorporation of two 18O atoms into the carboxyl terminus of each new peptide formed by cleavage of the adenovirus proteome. In the present study, a second enzyme, endoprotease Glu-C, is evaluated as an agent for cleavage and labeling. Proteolytic 18O labeling by Glu-C is shown to occur readily with phosphorylated and glycosylated proteins and with cysteinealkylated and disulfide-linked proteins. A sequential double-labeling strategy is used to characterize N-linked glycopeptides. Labeled and unlabeled peptide pairs are found to coelute chromatographically, and measurements of isotope ratios by nanospray and capillary LC-MS are found to be accurate and precise. PMID- 12643525 TI - A new isoelectric focusing gel for two-dimensional electrophoresis constructed in microporous hollow fiber membranes. AB - We describe the preparation of IEF tube gels inside a nonwetting microporous plastic tubing. The gel in the tube need not be extruded after the first dimension separation. Instead, the porous structure of the tubes is made wettable, and the proteins are electrophoresed "through-the-wall" into the second dimension PAGE gel. Commercial ampholytes and reagents are suitable for the procedure. A useful p/ range of 4.5-9.5 can be obtained when p/ 3-10 ampholyte mixtures are used. Because of the high surface area of the porous material, precautions must be exercised to reduce oxygen inhibition during polymerization and dehydration of the gel during storage and use. A sheath device is described that satisfies these requirements. The plastic tubes can be disposed of by incineration and pose no biohazard. PMID- 12643527 TI - Prediction of treatment response using gene expression profiles. AB - This paper concerns prediction of clinical outcome from gene expression profiles using work in a different area, nonlinear system identification. In particular, the approach can predict long-term treatment response from data of a landmark article by Golub et al. (Golub, T. R.; Slonim, D. K.; Tamayo, P.; Huard, C.; Gaasenbeek, M.; Mesirov, J. P. et al. Science 1999, 286, 531-537) that has not previously been achieved with these data. The present paper shows that, for these data, gene expression profiles taken at time of diagnosis of acute myeloid leukemia contain information predictive of eventual response to chemotherapy. This was not evident in previous work; indeed, the Golub et al. article did not find a set of genes strongly correlated with clinical outcome. However, the present approach can accurately predict outcome class of gene expression profiles even when the genes do not have large differences in expression levels between the classes. PMID- 12643526 TI - Proteome analysis of low-abundance proteins using multidimensional chromatography and isotope-coded affinity tags. AB - The effectiveness of proteome-wide protein identification and quantitative expression profiling is dependent on the ability of the analytical methodologies employed to routinely obtain information on low-abundance proteins, as these are frequently of great biological importance. Two-dimensional gel electrophoresis, the traditional method for proteome analysis, has proven to be biased toward highly expressed proteins. Recently, two-dimensional chromatography of the complex peptide mixtures generated by the digestion of unseparated protein samples has been introduced for the identification of their components, and isotope-coded affinity tags (ICAT) have been introduced to allow for accurate quantification of the components of protein mixtures by mass spectrometry. Here, we demonstrate that the combination of isotope coded affinity protein tags and multidimensional chromatography/mass spectrometry of tryptic peptide mixtures is capable of detecting and quantifying proteins of low abundance in complex samples. PMID- 12643528 TI - Peptide end sequencing by orthogonal MALDI tandem mass spectrometry. AB - Highly sensitive peptide fragmentation and identification in sequence databases is a cornerstone of proteomics. Previously, a two-layered strategy consisting of MALDI peptide mass fingerprinting followed by electrospray tandem mass spectrometry of the unidentified proteins has been successfully employed. Here, we describe a high-sensitivity/high-throughput system based on orthogonal MALDI tandem mass spectrometry (o-MALDI) and the automated recognition of fragments corresponding to the N- and C-terminal amino acid residues. Robotic deposition of samples onto hydrophobic anchor substrates is employed, and peptide spectra are acquired automatically. The pulsing feature of the QSTAR o-MALDI mass spectrometer enhances the low mass region of the spectra by approximately 1 order of magnitude. Software has been developed to automatically recognize characteristic features in the low mass region (such as the y1 ion of tryptic peptides), maintaining high mass accuracy even with very low count events. Typically, the sum of the N-terminal two ions (b2 ion), the third N-terminal ion (b3 ion), and the two C-terminal fragments of the peptide (y1 and y2) can be determined. Given mass accuracy in the low ppm range, peptide end sequencing on one or two tryptic peptides is sufficient to uniquely identify a protein from gel samples in the low silver-stained range. PMID- 12643530 TI - Proteomic priorities. PMID- 12643531 TI - Near-field optical analysis of living cells in vitro. AB - Near-field optical analysis (NOA) provides morphological nanoscale mappings of living cells in liquid cell culture media and nondestructive insight into cell functionality. Here we show for the first time the performance of NOA in imaging living cells. Unlabeled human endothelial cells attached to polished titanium disks were analyzed with hydrophobically coated optical biosensors mounted to a near-field scanning optical microscope (NSOM). Biosensors and titanium substrates could be simply implemented in standard NSOM and high-throughput NOA. PMID- 12643529 TI - Proteomic analysis of rat kidney cortex following treatment with gentamicin. AB - The regionally specific structure and function of the kidney renders it susceptible to toxic exposure. To characterize these changes at the proteome level, we have investigated the effects on protein expression following treatment with gentamicin. The more than 20 proteins identified were involved in the citric acid cycle, gluconeogenesis, fatty acid synthesis, and transport or cellular stress responses. These results strongly support the notion that energy production is impaired and mitochondrial dysfunction is involved in gentamicin induced nephrotoxicity. PMID- 12643532 TI - Studies in the assessment of folding quality for protein modeling and structure prediction. AB - A diagnostic for assessing the quality of a fold has been developed to which further criteria can be progressively added. The goal is to create a measure that can follow the status of a protein structure in a simulation or modeling process, when the answer (the experimental structure) is not known in advance, rather than simply reject deliberate misfolds. This places greater emphasis on the need to study, and calibrate against, marginal cases, i.e., unusual native structures, incomplete structures, partially erroneous X-ray structures, good models, poor models, and the effect of cofactors. The first three terms introduced in the diagnostic are appropriate core-forming properties or noncore properties of residues in relation to tertiary structure, appropriate neighboring structure density for each residue in relation to tertiary structure, and secondary structure consistency. While the method emerges as a useful simulation analysis tool, we find a need for further fine-tuning to diminish sensitivity to minor conformational changes that retain essential features of the fold, balanced against the need to obtain a more sensitive response when a conformational change involves less physically meaningful interatomic interactions. This dual utility is difficult to obtain: the investigation highlights some of the issues. Initial attempts to obtain it have led to terms in the diagnostic that are admittedly complex: simplifications must also be explored. PMID- 12643533 TI - Proteomic 2DE database for spot selection, automated annotation, and data analysis. AB - We present a software solution that enables faster and more accurate data analysis of 2DE/MALDI TOF MS data. The software supports data analysis through a number of automated data selection functions and advanced graphical tools. Once protein identities are determined using MALDI TOF MS, automated data retrieval from online databases provides biological information. The software, called 2DDB, reduces analysis time to a fraction without losing any quality compared to more manual data analysis. The database contains over 100,000 data entries, and selected parts can be reached at http://2ddb.org. PMID- 12643534 TI - Minimizing resolution of isotopically coded peptides in comparative proteomics. AB - Stable isotopes are now widely used to quantify concentration changes in proteomics. This paper focuses on the resolution of isotopically coded peptides and how isotope effects occurring during chromatographic separations can be minimized. Heavy isotope derivatizing agents used in this work were the commercially available 2H8-ICAT reagent and 13C4-succinic anhydride. The ICAT reagent derivatizes cysteine-containing peptides, whereas the succinic anhydride reacts with primary amine groups in peptides. It was observed during reversed phase chromatography of peptides from a BSA tryptic digest differentially labeled with the 2Hr and 2H8-ICAT reagents that resolution of the isoforms exceeded 0.5 with 20% of the peptides in the digest. Three-fourths of the peptides in this group contained two cysteine residues and were doubly labeled. Only 23% of the peptides labeled with a single ICAT residue had a resolution greater than 0.4. The resolution of peptides differentially labeled with 13C- and 12C-succinate never exceeded +/- 0.01, even in the case of peptides from the BSA digest labeled with 2 mol of succinate. Because this value is within the limits of the method used to determine resolution, it was concluded the 13C- and 12C-coded isoforms of labeled peptides did not resolve. The isotope ratio in the case of 13C/12C coding could be determined from a single mass spectrum taken at any point in the elution profile. This enabled isotope ratio analysis to be completed early in the elution of a peptide from chromatography columns. PMID- 12643535 TI - Effect of zinc and temperature on the conformation of the gamma subunit of retinal phosphodiesterase: a natively unfolded protein. AB - The cyclic GMP phosphodiesterase gamma-subunit (PDEgamma) was shown to belong to the family of natively unfolded proteins. Increasing temperature transforms the protein into a more ordered (but still relatively disordered) conformation. The C terminal part of PDEgamma has a high-affinity zinc-binding site (Kd approximately 1 microM), with His75 and His79 being directly involved into the coordination of Zn2+. Zinc-loaded protein remains effectively unfolded. Possible implications of these findings to the functioning of PDEgamma are discussed. PMID- 12643536 TI - Biomarker discovery in urine by proteomics. AB - A hypothesis was formed that it would be possible to isolate an adequate amount of protein from a patient, having normal renal function, to identify biological markers of a particular disease state using a variety of proteomics techniques. To support this hypothesis, three samples of urine were collected from a volunteer: first when healthy, later when experiencing acute inflammation due to a pilonidal abcess, and again later still after successful recovery from the condition. The urine from these samples was processed by solid-phase extraction to concentrate and desalt the endogenous proteins and peptides. The proteins and peptides from these urine samples were analyzed in three different experiments: (1) traditional two-dimensional gel electrophoresis followed by proteolysis and mass spectrometric identification of various protein spots, (2) whole mixture proteolysis followed by one-dimensional packed capillary liquid chromatography and tandem mass spectrometry, (3) whole mixture proteolysis followed by two dimensional capillary liquid chromatography and tandem mass spectrometry. In all three cases, a set of proteins was identified representing putative biomarkers. Each of these proteins was then found to have been previously linked in the scientific literature to inflammation. One acute phase reactant in particular, orosomucoid, was readily observed in all three experiments to dramatically increase in abundance, thereby supporting the hypothesis. PMID- 12643537 TI - Off-line coupling of high-resolution capillary electrophoresis to MALDI-TOF and TOF/TOF MS. AB - High-resolution capillary electrophoresis has been coupled to MALDI-TOF and TOF/TOF MS through off-line vacuum deposition onto standard stainless steel MALDI targets. This off-line approach allowed the decoupling of the separation from the MS analysis, thus allowing each to be independently optimized in terms of time. Using BSA tryptic digest as a model sample, the deposited streaks, roughly 100 microm wide, were first analyzed in the MS mode, consuming only a fraction of the sample. After data analysis, segments of the deposited trace, containing unidentified peptides, as well as several species chosen for sequence confirmation, were reanalyzed in the MS/MS mode using MALDI-TOF/TOF MS. Additionally, it is shown that the shot-to-shot reproducibility of the vacuum deposited trace (5% RSD) is 1 order of magnitude lower than that found for the standard dried droplet method. Moreover, a linear dependence of signal intensities (relative to an internal standard) over 3 orders of magnitude was found for a peptide sample with concentrations ranging from 1 to 1000 nM. This paper demonstrates the potential of off-line coupling of high-resolution separations to MALDI-MS and MALDI-MS/MS using vacuum deposition for the analysis of complex peptide mixtures from protein digests. PMID- 12643538 TI - Cyclization of N-terminal S-carbamoylmethylcysteine causing loss of 17 Da from peptides and extra peaks in peptide maps. AB - Enzymatic digests of proteins S-alkylated with iodoacetamide may contain peptides with N-terminal S-carbamoylmethylcysteine. These can be partly converted to a form with 17 Da lower mass and increased HPLC retention. Proof by synthesis supported by MS/MS and NMR spectroscopy was used to show that N-terminal S carbamoylmethyl-L-cysteine can cyclize, losing NH3 to form an N-terminal residue of (R)-5-oxoperhydro-1,4-thiazine-3-carboxylic acid. The abbreviation Otc is proposed for the (R)-5-oxoperhydro-1,4-thiazine-3-carbonyl residue. The rate of cyclization is significant in 0.1 M NH4HCO3 at 37 degrees C, with the half-life of the acyclic form being 10-12 h for several peptides tested. This is similar to the rate at which N-terminal pyroglutamate forms from N-terminal glutamine. PMID- 12643539 TI - Quantitation using enhanced signal tags: a technique for comparative proteomics. AB - Differential amidination of N-termini and lysine residues provides the basis for a novel approach to protein quantitation using MALDI mass spectrometry. Because the amidination of lysines increases their basicity and therefore MALDI ionization yields, the method is called quantitation using enhanced signal tags (QUEST). Amidine labels differ by methylene groups, leading to 14 Da mass differentials. The utility of QUEST is demonstrated while analyzing the digests of two model proteins using MALDI-TOF mass spectrometry. PMID- 12643540 TI - Large-scale identification of Caenorhabditis elegans proteins by multidimensional liquid chromatography-tandem mass spectrometry. AB - A proteome of a model organism, Caenorhabditis elegans, was analyzed by an integrated liquid chromatography (LC)-based protein identification system, which was constructed by microscale two-dimensional liquid chromatography (2DLC) coupled with electrospray ionization (ESI) tandem mass spectrometry (MS/MS) on a high-resolution hybrid mass spectrometer with an automated data analysis system. Soluble and insoluble protein fractions were prepared from a mixed growth phase culture of the worm C. elegans, digested with trypsin, and fractionated separately on the 2DLC system. The separated peptides were directly analyzed by on-line ESI-MS/MS in a data-dependent mode, and the resultant spectral data were automatically processed to search a genome sequence database, wormpep 66, for protein identification. The total number of proteins of the composite proteome identified in this method was 1,616, including 110 secreted/targeted proteins and 242 transmembrane proteins. The codon adaptation indices of the identified proteins suggested that the system could identify proteins of relatively low abundance, which are difficult to identify by conventional 2D-gel electrophoresis (GE) followed by an offline mass spectrometric analysis such as peptide mass fingerprinting. Among the approximately 5,400 peptides assigned in this study, many peptides with post-translational modifications, such as N-terminal acetylation and phosphorylation, were detected. This expression profile of C. elegans, containing 571 hypothetical gene products, will serve as the basic data of a major proteome set expressed in the worm. PMID- 12643541 TI - Biophysical constraints for protein structure prediction. AB - Though highly desirable, neither a single experimental technique nor a computational approach can be sufficient enough to rationalize a protein structure. The incorporation of biophysical constraints, which can be rationalized based on conventional biophysical measurements, might lead to considerable improvement of the simulation procedures. In this regard, our analysis of 180 proteins in different conformational states allows prediction of the overall protein dimension based on the chain length, i.e., the protein molecular weight, with an accuracy of 10%. PMID- 12643542 TI - Evaluation of multidimensional chromatography coupled with tandem mass spectrometry (LC/LC-MS/MS) for large-scale protein analysis: the yeast proteome. AB - Highly complex protein mixtures can be directly analyzed after proteolysis by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). In this paper, we have utilized the combination of strong cation exchange (SCX) and reversed-phase (RP) chromatography to achieve two-dimensional separation prior to MS/MS. One milligram of whole yeast protein was proteolyzed and separated by SCX chromatography (2.1 mm i.d.) with fraction collection every minute during an 80 min elution. Eighty fractions were reduced in volume and then re-injected via an autosampler in an automated fashion using a vented-column (100 microm i.d.) approach for RP-LC-MS/MS analysis. More than 162,000 MS/MS spectra were collected with 26,815 matched to yeast peptides (7,537 unique peptides). A total of 1,504 yeast proteins were unambiguously identified in this single analysis. We present a comparison of this experiment with a previously published yeast proteome analysis by Yates and colleagues (Washburn, M. P.; Wolters, D.; Yates, J. R., III. Nat. Biotechnol. 2001, 19, 242-7). In addition, we report an in-depth analysis of the false-positive rates associated with peptide identification using the Sequest algorithm and a reversed yeast protein database. New criteria are proposed to decrease false-positives to less than 1% and to greatly reduce the need for manual interpretation while permitting more proteins to be identified. PMID- 12643544 TI - Multiple enzymatic digestion for enhanced sequence coverage of proteins in complex proteomic mixtures using capillary LC with ion trap MS/MS. AB - This study uses multiple enzyme digests to increase the sequence coverage of proteins identified by the shotgun sequencing approach to proteomic analysis. The enzymes used were trypsin, Lys-C, and Asp-N, which cleave at arginine and lysine residues, lysine, and aspartic acid residues, respectively. This approach was evaluated with the glycoprotein, tissue plasminogen activator, t-PA and gave enhanced sequence coverage, compared with a single enzymatic digest. The approach was then evaluated with a complex proteomic sample, namely plasma. It was found that trypsin and Lys-C were able to detect overlapping but distinct sets of proteins and a digital recombination of the data gave a significant increase in both the number of protein identifications as well as an increase in the number of peptides identified per protein (which improves the certainty of the assignment). PMID- 12643543 TI - Recoverin is a zinc-binding protein. AB - Recoverin is an N-myristoylated 23 kDa calcium-binding protein from retina, which modulates the Ca2+-sensitive deactivation of rhodopsin via Ca2+-dependent inhibition of rhodopsin kinase. It was shown by intrinsic and bis-ANS probe fluorescence, circular dichroism, and differential scanning calorimetry that myristoylated recombinant recoverin interacts specifically with zinc ions. Similar to the calcium binding, the binding of zinc to Ca2+-loaded recoverin additionally increases its alpha-helical content, hydrophobic surface area, and environmental mobility/polarity of its tryptophan residues. In contrast to the calcium binding, the binding of zinc decreases thermal stability of the Ca2+ loaded protein. Zn2+-titration of recoverin, traced by bis-ANS fluorescence, reveals binding of a single Zn2+ ion per protein molecule. It was shown that the double-mutant E85Q/E121Q with inactivated Ca2+-binding EF-hands 2 and 3 (Alekseev, A. M.; Shulga-Morskoy, S. V.; Zinchenko, D. V.; Shulga-Morskaya, S. A.; Suchkov, D. V.; Vaganova, S. A.; Senin, I. I.; Zargarov, A. A.; Lipkin, V. M.; Akhtar, M.; Philippov, P. P. FEBS Lett. 1998, 440, 116-118), which can be considered as an analogue of the apo-protein, binds Zn2+ ion as well. Apparent zinc equilibrium binding constants evaluated from spectrofluorimetric Zn2+ titrations of the protein are 1.4 x 10(5) M(-1) (dissociation constant 7.1 microM) for Ca2+-loaded wild-type recoverin and 3.3 x 10(4) M(-1) (dissociation constant 30 microM) for the E85Q/E121Q mutant (analogue of apo-recoverin). Study of the binding of wild-type recoverin to ROS membranes showed a zinc-dependent increase of its affinity for the membranes, without regard to calcium content, suggesting further solvation of a protein myristoyl group upon Zn2+ binding. Possible implications of these findings to the functioning of recoverin are discussed. PMID- 12643545 TI - Proteomic analysis of early melanosomes: identification of novel melanosomal proteins. AB - Melanin is a heterogeneous biopolymer produced only by specific cells termed melanocytes, which synthesize and deposit the pigment in specialized membrane bound organelles known as melanosomes. Although melanosomes have been suspected of being closely related to lysosomes and platelets, the total number of melanosomal proteins is still unknown. Thus far, six melanosome-specific proteins have been identified, and the challenge is to characterize the complete proteome of the melanosome to further understand its mechanism of biogenesis. In this report, we used mass spectrometry and subcellular fractionation to identify protein components of early melanosomes. Using this approach, we have identified all 6 of the known melanosome-specific proteins, 56 proteins that are shared with other organelles, and confirmed the presence of 6 novel melanosomal proteins using western blotting and by immunohistochemistry. PMID- 12643546 TI - Capillary affinity electrophoresis for the screening of post-translational modification of proteins with carbohydrates. AB - Glycosylation is one of the most important post-translational events for proteins, affecting their functions in health and disease, and plays significant roles in various information traffics for intracellular and intercellular biological events (Hancock, W. S. J. Proteome Res. 2002, 1, 297). We have attempted to obtain the information on the numbers and amounts of carbohydrate chains. Interaction between carbohydrate chains and proteins that recognize them is a target to understand the biological roles of glycosylation. To date, there have been a few strategies for simultaneous analysis of the interactions between complex mixtures of carbohydrates and proteins. Here, we report an approach to categorize carbohydrate chains using a few glycoprotein samples as models for the studies on the analysis of post-translational modification of proteins with carbohydrates. A combination of some specific lectins was used as carbohydrate binding proteins. The method is based on high-resolution separation of fluorescent-labeled carbohydrates by capillary electrophoresis with laser-induced fluorescent detection in the presence of carbohydrate-binding proteins at different concentrations. The present technique affords (1) simultaneous determination of carbohydrate chains, (2) binding specificity of the constituent carbohydrate chains to specific proteins, and (3) kinetic data such as the association constant of each carbohydrate. We found that the lectins employed in the present study could discriminate subtle difference in linkages and resolved the carbohydrate mixtures. The results will be useful, for example, to understand the biological events expressed with carbohydrate changes on the cell surface. PMID- 12643547 TI - Solubilization of trichloroacetic acid (TCA) precipitated microbial proteins via naOH for two-dimensional electrophoresis. AB - In preparing intracellular microbial samples for one- or two-dimensional electrophoresis, trichloroacetic acid (TCA) precipitation is frequently used to remove interfering compounds. Solubilization of TCA precipitate typically requires the addition of a number of chaotropes or detergents, in a multistep process, that requires hours to carry out. In this study, a simple, rapid, one step method to solubilize TCA precipitated proteins is presented. Precipitated proteins are pretreated with 0.2 M NaOH for less than 5 min, followed by addition of standard sample solubilization buffer (SSSB). When compared to solubilization with SSSB alone, NaOH pretreatment of TCA-precipitated intracellular protein from Aspergillus oryzae and Escherichia coli shows an approximate 5-fold increase in soluble protein. In addition, two-dimensional gel electrophoresis on resolubilized proteins shows an equivalent number of proteins in samples with and without NaOH pretreatment. PMID- 12643548 TI - Laser capture microdissection MALDI for direct analysis of archival tissue. AB - MALDI mass spectra were obtained from cancer cells isolated by laser capture microdissection (LCM) of archived tissue. Frozen human lung tissue from adenocarcenoma and squamous cell carcenoma cases were cut into 5 to 15 microm thick sections, stained with hematoxylin and dehydrated. Cancer cells were isolated by LCM, mixed with matrix solution, and deposited on a MALDI target for mass spectrometric analysis. For comparison with LCM isolated cells, tissue sections were placed directly on the MALDI target without microdissection. Tissue sections frozen in optimal cutting temperature (OCT) solution and cut into 8 microm thick sections gave the best performance with direct MALDI analysis. Between 15 and 20 peaks were observed in the mass region between 1,000 and 4,000 Da, and roughly half of these peaks were common to either squamous cells or adenocarcenoma. Additional peaks were observed in the non-LCM mass spectra and these may result from biomolecules in the healthy tissue. When compared to fresh tissue, both LCM and non-LCM archived tissue produced fewer peaks, possibly due to degradation of the biomolecules in the archived tissue. PMID- 12643549 TI - Radiologic image registration: old skills and new tools. PMID- 12643551 TI - Alignment of CT lung volumes with an optical flow method. AB - RATIONALE AND OBJECTIVES: This study was performed to evaluate an optical flow method for registering serial computed tomographic (CT) images of lung volumes to assist physicians in visualizing and assessing changes between CT scans. MATERIALS AND METHODS: The optical flow method is a coarse-to-fine model-based motion estimation technique for estimating first a global parametric transformation and then local deformations. Five serial pairs of CT images of lung volumes that were misaligned because of patient positioning, respiration, and/or different fields of view were used to test the method. RESULTS: Lung volumes depicted on the serial paired images initially were correlated at only 28%-68% because of misalignment. With use of the optical flow method, the serial images were aligned to at least 95% correlation. CONCLUSION: The optical flow method enables a direct comparison of serial CT images of lung volumes for the assessment of nodules or functional changes in the lung. PMID- 12643550 TI - Coregistration of head CT comparison studies: assessment of clinical utility. AB - RATIONALE AND OBJECTIVES: The authors evaluated the clinical utility of image coregistration in the interpretation of follow-up computed tomographic (CT) studies of the head. MATERIALS AND METHODS: Fourteen patients with 34 intracranial lesions underwent follow-up head CT. The images were coregistered automatically with proprietary software on a standard personal computer, and all patient demographic data were removed. A neuroradiologist read the coregistered images several days after first reading the nonregistered images. The reading was repeated some months later to assess intraobserver variability, and a second reader was recruited so that interobserver variability also could be assessed. The interpretations of nonregistered images served as controls for the interpretations of coregistered images. Readers were asked to assess changes in lesion size quantitatively and to record the time it took to evaluate each case. Differences in interpretation speed were evaluated with the t test. Univariate analysis was used to measure accuracy; interpretations were compared with those of a nonblinded senior neuroradiologist, which served as the diagnostic standard. Intra- and interindividual variability were assessed with the kappa statistic. RESULTS: The time needed to read the studies decreased by an average of 65.6% (P < .05), with statistically significant reductions for each reader. Coregistration also changed the interpretation results in 21.9% of cases. Coregistration increased the accuracy of reading, but not significantly. Intraobserver variability improved from 0.3554 to 0.7328 with coregistration, and interobserver variability improved from 0.2670 to 0.3309. CONCLUSION: Image coregistration is technically feasible and accurate. Coregistration of follow-up studies significantly reduces the time needed for comparison and interpretation. It does not detract from the accuracy of interpretation of follow-up studies and tends to decrease intra- and interobserver variability. PMID- 12643553 TI - Process utility for imaging in cerebrovascular disease. AB - RATIONALE AND OBJECTIVES: The morbidity associated with a diagnostic test can influence its cost-effectiveness, but the quantification of that morbidity is controversial. Accounting for pain and invasiveness requires the measurement of "process utility" in addition to the expected value of testing. An original time trade-off variant was applied to the imaging evaluation of cerebrovascular disease, for which differences in morbidity are important to patients. MATERIALS AND METHODS: A "waiting trade-off" (WTO) was used to evaluate the preferences of 89 patients for magnetic resonance (MR) angiography and conventional x-ray angiography. Patients were experienced with both tests. A weighted difference was calculated between the period a patient was willing to wait for a test result and treatment after a hypothetical "ideal" test and the choice to undergo conventional angiography or MR angiography with immediate treatment. A rating scale was used to check the convergent validity of the WTO. RESULTS: Paired data showed a highly significant difference (P = .0001) between the mean preference for conventional and MR angiography, favoring the latter and translating into a difference of 5 quality-adjusted life days. The more negatively patients judged their conventional angiographic experience, the longer they were willing to wait for the ideal test result. CONCLUSION: The WTO provides a reasonable estimate of the relative morbidity of more invasive conventional angiographic procedures and provides a quality-adjustment term for economic analysis. Such an approach may enable more complete evaluation of the effects of other processes on medical care. PMID- 12643552 TI - Establishing a normative atlas of the human lung: intersubject warping and registration of volumetric CT images. AB - RATIONALE AND OBJECTIVES: To establish the range of normal values for quantitative CT-based measures of lung structure and function, the authors developed a method for matching pulmonary structures across individuals and creating a normative human lung atlas. MATERIALS AND METHODS: A computerized human lung atlas was synthesized from computed tomographic (CT) images from six subjects by means of three-dimensional image registration. The authors identified a set of reproducible feature points for each CT image and used these points to establish correspondences across subjects, used a landmark- and intensity-based consistent image registration algorithm to register a template image volume from the population to the rest of the pulmonary CT volumes in the population, averaged these transformations, and constructed an atlas by deforming the template with the average transformation. RESULTS: The effectiveness of the authors' method was evaluated and visualized by means of both gray-level and segmented CT images. The method reduced the average landmark registration error from 10.5 mm to 0.4 mm and the average relative volume overlap error from 0.7 to 0.11 for the six data sets studied. CONCLUSION: The method, and the computerized human lung atlas constructed and visualized by the authors with this method, provides a basis for establishing regional ranges of normative values for structural and functional measures of the human lung. PMID- 12643554 TI - Analysis of a computer-assisted technique for measuring the lumbar spine on radiographs: correlation of two methods. AB - RATIONALE AND OBJECTIVES: The authors performed this study to compare the effectiveness of two methods used to measure intervertebral space height and sagittal vertebral translation. MATERIALS AND METHODS: Forty-five series of lateral lumbar spine radiographs were evaluated with two methods: distortion corrected radiographic analysis (DCRA) and computer-assisted lumbar spine measurement (CALSM). Intervertebral space height and sagittal translation depicted on radiographs obtained before and after the patients underwent spinal fusion were measured with both methods. The values obtained with CALSM were then correlated with those obtained with DCRA. RESULTS: The values of intervertebral space height and sagittal vertebral translation and of changes in both over time as assessed with CALSM did not show sufficient correlation with the values obtained with DCRA. Correlation coefficients ranged from -0.088 to 0.605 and were statistically highly significant (P < or = .01) for only six of the 48 items measured with both CALSM and DCRA. Moreover, without additional graphic constructs, CALSM provided sufficiently consistent identification of vertebral corner points in only 50% of cases. CONCLUSION: With regard to the measurement of intervertebral space height and sagittal translation on lateral lumbar spine radiographs, DCRA appears to be more reliable than CALSM. PMID- 12643555 TI - Performance change of mammographic CAD schemes optimized with most-recent and prior image databases. AB - RATIONALE AND OBJECTIVES: The authors evaluated performance changes in the detection of masses on "current" (latest) and "prior" images by computer-aided diagnosis (CAD) schemes that had been optimized with databases of current and prior mammograms. MATERIALS AND METHODS: The authors selected 260 pairs of matched consecutive mammograms. Each current image depicted one or two verified masses. All prior images had been interpreted originally as negative or probably benign. A CAD scheme initially detected 261 mass regions and 465 false-positive regions on the current images, and 252 corresponding mass regions (early signs) and 471 false-positive regions on prior images. These regions were divided into two training and two testing databases. The current and prior training databases were used to optimize two CAD schemes with a genetic algorithm. These schemes were evaluated with two independent testing databases. RESULTS: The scheme optimized with current images produced areas under the receiver operating characteristic curve of (0.89 +/- 0.01 and 0.65 +/- 0.02 when tested with current images and prior images, respectively. The scheme optimized with prior images produced areas under the receiver operating characteristic curve of 0.81 +/- 0.02 and 0.71 +/- 0.02 when tested with current images and prior images, respectively. Performance changes for both current and prior testing databases were significant (P < .01) for the two schemes. CONCLUSION: CAD schemes trained with current images do not perform optimally in detecting masses depicted on prior images. To optimize CAD schemes for early detection, it may be important to include in the training database a large fraction of prior images originally reported as negative and later proven to be positive. PMID- 12643556 TI - Routine editing of trainee-generated radiology reports: effect on style quality. AB - RATIONALE AND OBJECTIVES: The authors performed this study to determine the effect of routine editing on the style quality of trainee-generated radiology reports. MATERIALS AND METHODS: Trainee-generated reports of 50 body computed tomographic scans obtained at a tertiary care cancer center were edited in a routine fashion by one of two attending radiologists. Three physicians and four radiologists each independently evaluated the randomized unedited and edited reports (n = 100) and rated each report for clarity, brevity, readability, and quality of the impression by using a five-point scale. RESULTS: Editing significantly improved mean ratings for clarity (4.6 after editing vs 4.2 before editing, P < .007), brevity (4.6 vs 4.2, P < .007), readability (4.4 vs 4.1, P < .007), and quality of the impression (4.5 vs 4.3, P < .007). CONCLUSION: Routine editing of trainee-generated reports significantly improves the perceived report quality. This finding suggests that greater emphasis should be placed on stylistic aspects of reporting during training to improve report style quality at dictation. PMID- 12643557 TI - Medical student radiology training: what are the objectives for contemporary medical practice? AB - RATIONALE AND OBJECTIVES: The authors attempt to provide a set of objectives for medical student training in radiology for contemporary medical practice. MATERIALS AND METHODS: A questionnaire containing a list of educational objectives was sent to 32 radiologists in charge of medical student training in radiology at accredited residency programs in Australia and New Zealand. The importance of including each preselected objective in the curriculum was measured by respondents' agreement or disagreement on a scale of 1-6. Opportunity also was given to respondents to suggest objectives other than those presented on the questionnaire. RESULTS: Twenty of the 32 questionnaires were returned, and a set of educational objectives was established based on the responses. The objectives were ranked in importance according to the mean score assigned to each objective by the respondents. CONCLUSION: This new set of educational objectives for medical student radiology training reflects recent changes in radiologic and medical practice and points to potential future developments. PMID- 12643558 TI - Commentary on learning to report. PMID- 12643559 TI - The radiology fellowship and fellowship match: current status. PMID- 12643560 TI - Three-dimensional volume rendering of fetal MR images for the diagnosis of congenital cystic adenomatoid malformation. AB - RATIONALE AND OBJECTIVES: Great expertise is necessary to mentally compile a series of individual two-dimensional image sections into a three-dimensional (3D) composite view that can aid in differential diagnosis. The purpose of this study was to test 3D volume-rendering techniques for differentiating congenital cystic adenomatoid malformation from congenital diaphragmatic hernia. MATERIALS AND METHODS: The authors acquired T2-weighted magnetic resonance (MR) images of a 27 week fetus in the sagittal plane and then applied the 3D volume-rendering method to the MR image data sets to obtain a composite 3D image. RESULTS: It was unclear on the MR images whether the intestines were situated above or below the diaphragm. The composite image showed that the intestines were not herniated into the chest, and this facilitated a diagnosis of congenital cystic adenomatoid malformation rather than congenital diaphragmatic hernia. CONCLUSION: The 3D volume-rendering techniques aided in the assessment of fetal organ structure and could be applied also to preoperative simulation and planning of fetal surgery. PMID- 12643562 TI - Economic, legal, and ethical rationales for the ACRIN national lung screening trial of CT screening for lung cancer. PMID- 12643561 TI - CO2-enhanced CT imaging for the detection of pulmonary emboli: feasibility study in a porcine model. AB - RATIONALE AND OBJECTIVES: The authors investigated the feasibility of using computed tomography (CT) with CO2 gas as a negative contrast agent for detecting pulmonary emboli in a porcine model. MATERIALS AND METHODS: Seven pigs with or without pulmonary emboli underwent thoracic imaging with multi-detector row spiral CT. To identify optimal injection and scanning protocols, the first four pigs were scanned repeatedly in the supine and prone positions with different scan delays (10, 15, and 20 seconds) and different volumes of CO2 (60, 120, 180, and 240 mL), which were hand infused (each infusion took 10-15 seconds). The last five pigs with emboli were scanned with iodinated contrast medium and then rescanned with 120 or 180 mL of CO2. The CO2 volumes and scan delays were qualitatively assessed. The supine and prone CT scans and the number and location of thrombi depicted in the CO2- and contrast material-enhanced CT scans were compared. RESULTS: Because the pulmonary artery in pigs is in the posterior anatomy, the prone position was more effective than the supine position with CO2 enhancement. An infusion of 120 mL of CO2 was sufficient to enhance the entire pulmonary artery, and scanning timed to coincide with the completion of infusion was the most effective. Both the CO2- and contrast-enhanced CT scans demonstrated all thrombi. Thrombi were more apparent on the CO2-enhanced CT scans than on the contrast-enhanced scans because of the high contrast interface between soft tissue and gas. However, two of the seven pigs with thrombi experienced abrupt cardiac arrest after CO2-enhanced scanning and could not be resuscitated. The cause of these events was not determined in the current study. CONCLUSION: The CT depiction of pulmonary emboli is feasible with CO2 gas as a negative contrast agent and may even be superior to that with iodinated contrast media. Further studies are required to evaluate the safety of this method and to develop an improved delivery of CO2 gas for this application. PMID- 12643563 TI - The adaptive dynamics of altruism in spatially heterogeneous populations. AB - We study the spatial adaptive dynamics of a continuous trait that measures individual investment in altruism. Our study is based on an ecological model of a spatially heterogeneous population from which we derive an appropriate measure of fitness. The analysis of this fitness measure uncovers three different selective processes controlling the evolution of altruism: the direct physiological cost, the indirect genetic benefits of cooperative interactions, and the indirect genetic costs of competition for space. In our model, habitat structure and a continuous life cycle makes the cost of competing for space with relatives negligible. Our study yields a classification of adaptive patterns of altruism according to the shape of the costs of altruism (with decelerating, linear, or accelerating dependence on the investment in altruism). The invasion of altruism occurs readily in species with accelerating costs, but large mutations are critical for altruism to evolve in selfish species with decelerating costs. Strict selfishness is maintained by natural selection only under very restricted conditions. In species with rapidly accelerating costs, adaptation leads to an evolutionarily stable rate of investment in altruism that decreases smoothly with the level of mobility. A rather different adaptive pattern emerges in species with slowly accelerating costs: high altruism evolves at low mobility, whereas a quasi-selfish state is promoted in more mobile species. The high adaptive level of altruism can be predicted solely from habitat connectedness and physiological parameters that characterize the pattern of cost. We also show that environmental changes that cause increased mobility in those highly altruistic species can beget selection-driven self-extinction, which may contribute to the rarity of social species. PMID- 12643564 TI - Phylogenetically nested comparisons for testing correlates of species richness: a simulation study of continuous variables. AB - Explaining the uneven distribution of species among lineages is one of the oldest questions in evolution. Proposed correlations between biological traits and species diversity are routinely tested by making comparisons between phylogenetic sister clades. Several recent studies have used nested sister-clade comparisons to test hypotheses linking continuously varying traits, such as body size, with diversity. Evaluating the findings of these studies is complicated because they differ in the index of species richness difference used, the way in which trait differences were treated, and the statistical tests employed. In this paper, we use simulations to compare the performance of four species richness indices, two choices about the branch lengths used to estimate trait values for internal nodes and two statistical tests under a range of models of clade growth and character evolution. All four indices returned appropriate Type I error rates when the assumptions of the method were met and when branch lengths were set proportional to time. Only two of the indices were robust to the different evolutionary models and to different choices of branch lengths and statistical tests. These robust indices had comparable power under one nonnull scenario. Regression through the origin was consistently more powerful than the t-test, and the choice of branch lengths exerts a strong effect on both the validity and power. In the light of our simulations, we re-evaluate the findings of those who have previously used nested comparisons in the context of species richness. We provide a set of simple guidelines to maximize the performance of phylogenetically nested comparisons in tests of putative correlates of species richness. PMID- 12643565 TI - An experimental test of local adaptation in soil bacteria. AB - We extracted bacterial isolates of similar colony morphology from spatially located soil samples within 1 ha of old-growth forest. The same soil samples were used to prepare growth medium. Each isolate was then cultured in each medium and its growth recorded. There was no overall tendency for isolates to grow more successfully in their home site (i.e., the medium derived from the soil sample from which they had been extracted). Most isolates grew very poorly, however, and when the analysis was restricted to the minority of vigorous isolates there was clear evidence of local adaptation: isolates tended to grow better at their home site than did isolates from elsewhere and grew better at their home site than they did at other sites. The variation of growth within the 1-ha plot made up a complex fitness landscape of peaks, ridges, and valleys. Most of the vigorous isolates were found at or near a local fitness (growth) peak, although seldom at a global peak. In consequence, there was a tendency for growth to diminish away from the home site. The home isolate was about 50% more fit than average at its home site; fitness diminished exponentially away from the home site at a rate of 0.0577 per meter. These figures are similar to those previously reported for plants. This selection gradient has matched the bacterial assemblage to the edaphic structure of the environment, although the fit is far from perfect. PMID- 12643566 TI - Molecular evidence for host-adapted races of the fungal endophyte Epichloe bromicola after presumed host shifts. AB - Host shifts of plant-feeding insects and parasites promote adaptational changes that may result in the formation of host races, an assumed intermediate stage in sympatric speciation. Here, we report on genetically differentiated and host adapted races of the fungal endophyte Epichloe bromicola, which presumably emerged after a shift from the grass Bromus erectus to other Bromus hosts. Fungi of the genus Epichloe (Ascomycota) and related anamorphs of Neotyphodium are widespread endophytes of cool-season grasses. Sexually reproducing strains sterilize the host by formation of external fruiting structures (stromata), whereas asexual strains are asymptomatic and transmitted via seeds. In E. bromicola, strains infecting B. erectus are sexual, and strains from two woodland species, B. benekenii and B. ramosus, are asexual and seed transmitted. Analyses of amplified fragment length polymorphism fingerprinting and of intron sequences of the tub2 and tef1 genes of 26 isolates from the three Bromus hosts collected at natural sites in Switzerland and nearby France demonstrated that isolates are genetically differentiated according to their host, indicating that E. bromicola does not form a single, randomly mating population. Phylogenetic analyses of sequence data did not unambiguously resolve the exact origin of asexual E. bromicola strains, but it is likely they arose from within sexual populations on B. erectus. Incongruence of trees derived from different genes may have resulted from recombination at some time in the recent history of host strains. Reciprocal inoculations of host plant seedlings showed that asexual isolates from B. benekenii and B. ramosus were incapable of infecting B. erectus, whereas the sexual isolates from B. erectus retained the assumed ancestral trait of broad compatibility with Bromus host seedlings. Because all isolates were interfertile in experimental crosses, asexual strains may not be considered independent biological species. We suggest that isolates infecting B. benekenii and B. ramosus represent long-standing host races or incipient species that emerged after host shifts and that may evolve through host-mediated reproductive isolation toward independent species. PMID- 12643567 TI - Evolutionary processes in the genus Coreocarpus: insights from molecular phylogenetics. AB - A molecular phylogenetic study of the plant genus Coreocarpus was conducted using nuclear (ITS) and plastid (rpl16 intron) DNA sequences, with phylogenies of the nuclear and plastid sequences highly congruent in defining a monophyletic group of six species (core Coreocarpus), although three other species often placed within the genus were excluded. Relationships within the genus are largely but not totally concordant with prior biosystematic studies. Despite strong molecular support, no morphological characters uniting the six species of core Coreocarpus have been identified; retention of plesiomorphic characters and the genetic lability of characters are two probable factors contributing to lack of consistent defining characters. The age of the core Coreocarpus is estimated at 1 million years because the basal species is endemic to a volcanic island that emerged in the past million years. Mapping the results of earlier breeding studies on the molecular phylogeny showed that use of cross-compatibility as a criterion for species delimitation would result in the recognition of paraphyletic species. Prior field, morphological, and biosystematic studies provided no indication of past hybridization in the evolution of Coreocarpus, and species in the genus appeared to be well defined morphologically. However, three instances of incongruence were observed. Two of these were between the nuclear and plastid partitions, and the third was between the morphological species assignment of one accession and the molecular data. If hybridization accounts for incongruence between the nuclear and plastid data, it occurred between species that now appear to be cross-incompatible and allopatric. The incongruence between morphological species assignment and the molecular data could be the result of parallel fixation of characters that have a simple genetic basis. This study suggests that the evolutionary history of Coreocarpus is much more complex than indicated from prior biosystematic investigations and that biosystematic and molecular phylogenetic studies may complement each other for elucidating the evolution and phylogeny of a group. PMID- 12643568 TI - Genetic structure of age classes in Camellia japonica (Theaceae). AB - Camellia japonica L. (Theaceae), an insect- and bird-pollinated, broad-leaved evergreen tree, is widely distributed in Japan and the southern Korean peninsula. The species has a relatively even age distribution within populations, which may influence the spatial genetic structure of different age classes relative to species with typical L-shaped age distributions. To determine whether the internal spatial genetic structure found in seedlings and young individuals carries over into adults, we used allozyme loci, F-statistics, spatial autocorrelation statistics (Moran's I), and coancestry measures to examine changes in genetic structure among seven age classes in a population (60-m x 100 m area) in southern Korea. In seedlings, weak but significant positive values of Moran's I-statistics and coancestry measures were found for distances less than 14 m, which is consistent with a mechanism of limited seed dispersal combined with overlapping seed shadows. This spatial structure, however, dissipates in older age classes, and in adults genetic variation has an essentially random spatial distribution. Morisita's index of dispersion of individuals in each age class showed that seedlings and juveniles are more highly clustered than are older individuals. These results suggest that self-thinning changes the spatial relationships of individuals, and thus genotypes. A multilocus estimate of FST (0.008) shows a small but statistically significant difference in allele frequencies among age classes. In summary, intrapopulation genetic structure within and among age classes of C. japonica was significant but weak. Despite presumably limited seed dispersal, weak spatial genetic structure in juveniles suggests overlapping seed shadows followed by self-thinning during recruitment. The present study also demonstrates that studies of spatial genetic structure focusing on limited numbers of generations may not be sufficient to reveal the entire picture of genetic structure in populations with overlapping generations. PMID- 12643569 TI - Floral morphometrics and the evolution of sexual dimorphism in Lycium (Solanaceae). AB - Plants of Lycium californicum, L. exsertum, and L. fremontii produce flowers that are either male-sterile (female) or hermaphroditic, and populations are morphologically gynodioecious. As is commonly found in gynodioecious species, flowers on female plants are smaller than those on hermaphrodites for a number of floral traits. Floral size dimorphism has often been hypothesized to be the result of either a reduction in female flower size that allows reallocation to greater fruit and seed production, or an increase in hermaphroditic flower size due to the increased importance of pollinator attraction and pollen export for hermaphroditic flowers. We provide a test of these two alternatives by measuring 11 floral characters in eight species of Lycium and using a phylogeny to reconstruct the floral size shifts associated with the evolution of gender dimorphism. Our analyses suggest that female flowers are reduced in size relative to the ancestral condition, whereas flowers on hermaphrodites have changed only slightly in size. Female and hermaphroditic flowers have also diverged both from one another and from ancestral cosexual species in several shape characteristics. We expected sexual dimorphism to be similar among the three dimorphic taxa, as gender dimorphism evolved only a single time in the ancestor of the American dimorphic lineage. While the floral sexual dimorphism is broadly similar among the three dimorphic species, there are some species-specific differences. For example, L. exsertum has the greatest floral size dimorphism, whereas L. fremontii had the greatest size-independent dimorphism in pistil characters. To determine the degree to which phylogenetic uncertainty affected reconstruction of ancestral character states, we performed a sensitivity analysis by reconstructing ancestral character states on alternative topologies. We argue that investigations such as this one, that examine floral evolution from an explicitly phylogenetic perspective, provide new insights into the study of the evolution of floral sexual dimorphism. PMID- 12643570 TI - Phylogenetic analysis of correlation structure in stalk-eyed flies (Diasemopsis, Diopsidae). AB - Morphological divergence among species may be constrained by the pattern of genetic variances and covariances among traits within species. Assessing the existence of such a relationship in nature requires information on the stability of intraspecific correlation and covariance structure and the correspondence of this structure to the pattern of evolutionary divergence within a lineage. Here, we investigate these issues for nine morphological traits and 15 species of stalk eyed flies in the genus Diasemopsis. Within-species matrices for these traits were generated from phenotypic data for all the Diasemopsis species and from genetic data for a single Diasemopsis species, D. dubia. The among-species pattern of divergence was assessed by calculating the evolutionary correlations for all pairwise combinations of the morphological traits along the phylogeny of these species. Comparisons of intraspecific matrices reveal significant similarity among all species in the phenotypic correlations matrices but not the covariance matrices. In addition, the differences in correlation structure that do exist among species are not related to their phylogenetic placement or change in the means of the traits. Comparisons of the phenotypic and phylogenetic matrices suggest a strong relationship between the pattern of evolutionary change among species and both the intraspecific correlation structure and the stability of this structure among species. The phenotypic and the phylogenetic matrices are significantly similar, and pairs of traits whose intraspecific correlations are more stable across taxa exhibit stronger coevolution on the phylogeny. These results suggest either the existence of strong constraints on the pattern of evolutionary change or a consistent pattern of correlated selection shaping both the phenotypic and phylogenetic matrices. The genetic correlation structure for D. dubia, however, does not correspond with patterns found in the phenotypic and phylogenetic data. Possible reasons for this disagreement are discussed. PMID- 12643571 TI - Adaptation of rainbow fish to lake and stream habitats. AB - Fish occupy a range of hydrological habitats that exert different demands on locomotor performance. We examined replicate natural populations of the rainbow fishes Melanotaenia eachamensis and M. duboulayi to determine if colonization of low-velocity (lake) habitats by fish from high-velocity (stream) habitats resulted in adaptation of locomotor morphology and performance. Relative to stream conspecifics, lake fish had more posteriorly positioned first dorsal and pelvic fins, and shorter second dorsal fin bases. Habitat dimorphism observed between wild-caught fish was determined to be heritable as it was retained in M. eachamensis offspring raised in a common garden. Repeated evolution of the same heritable phenotype in independently derived populations indicated body shape divergence was a consequence of natural selection. Morphological divergence between hydrological habitats did not support a priori expectations of deeper bodies and caudal peduncles in lake fish. However, observed divergence in fin positioning was consistent with a family-wide association between habitat and morphology, and with empirical studies on other fish species. As predicted, decreased demand for sustained swimming in lakes resulted in a reduction in caudal red muscle area of lake fish relative to their stream counterparts. Melanotaenia duboulayi lake fish also had slower sustained swimming speeds (Ucrit) than stream conspecifics. In M. eachamensis, habitat affected Ucrit of males and females differently. Specifically, females exhibited the pattern observed in M. duboulayi (lake fish had faster Ucrit than stream fish), but the opposite association was observed in males (stream males had slower Ucrit than lake males). Stream M. eachamensis also exhibited a reversed pattern of sexual dimorphism in Ucrit (males slower than females) relative to all other groups (males faster than females). We suggest that M. eachamensis males from streams responded to factors other than water velocity. Although replication of muscle and Ucrit phenotypes across same habitat populations within and/or among species was suggestive of adaptation, the common garden experiment did not confirm a genetic basis to these associations. Kinematic studies should consider the effect of the position and base length of dorsal fins. PMID- 12643572 TI - Molecular systematics, phylogeography, and the effects of Pleistocene glaciation in the painted turtle (Chrysemys picta) complex. AB - The painted turtle, Chrysemys picta, is currently recognized as a continentally distributed polytypic species, ranging across North America from southern Canada to extreme northern Mexico. We analyzed variation in the rapidly evolving mitochondrial control region (CR) in 241 turtles from 117 localities across this range to examine whether the painted turtle represents a continentally distributed species based on molecular analysis. We found strong support for the novel hypothesis that C. p. dorsalis is the sister group to all remaining Chrysemys, with the remaining Chrysemys falling into a single, extremely wide ranging and genetically undifferentiated species. Given our goal of an evolutionarily accurate taxonomy, we propose that two evolutionary lineages be recognized as species within Chrysemys: C. dorsalis (Agassiz 1857) in the southern Mississippi drainage region, and C. picta (Schneider 1783) from the rest of the range of the genus. Neither molecular nor recent morphological analyses argue for the hybrid origin of C. p. marginata as previously proposed. Within C. picta, we find evidence of at least two independent range expansions into previously glaciated regions of North America, one into New England and the other into the upper Midwest. We further find evidence of a massive extinction/recolonization event across the Great Plains/Rocky Mountain region encompassing over half the continental United States. The timing and extent of this colonization is consistent with a recently proposed regional aridification as the Laurentide ice sheets receded approximately 14,000 years ago, and we tentatively propose this paleoclimatological event as a major factor shaping genetic variation in Chrysemys. PMID- 12643573 TI - Dispersal of viviparity across contact zones in Iberian populations of fire salamanders (Salamandra) inferred from discordance of genetic and morphological traits. AB - We used partial sequences of the cytochrome b mitochondrial DNA (mtDNA) gene, obtained from 76 individuals representing 45 populations of Iberian Salamandra salamandra plus 15 sequences of additional species of Salamandra and related genera, to investigate contact zones. These zones, identified by earlier allozymic and morphological analyses, are between populations of viviparous (S. s. bernardezi and S. s. fastuosa) and ovoviviparous (S. s. gallaica and S. s. terrestris) salamanders. The distribution of mtDNA and nuclear markers is mostly concordant at one contact zone (between S. s. gallaica and S. s. bernardezi), but at another (between S. s. fastuosa and S. s. terrestris) the markers are offset by about 250 km. The observed geographic variation fits a model of mtDNA capture. Among the potential mechanisms responsible for such discordance we favor a combination of range shifts due to climatic fluctuations and biased genetic admixture across moving contact zones. We apply our findings to the issue of possible homoplasy in the evolution of viviparity and conclude that viviparity likely arose only once within S. salamandra. PMID- 12643574 TI - Effects of common origin and common environment on nestling plumage coloration in the great tit (Parus major). AB - Carotenoids cannot be synthesized by birds and thus have to be ingested with food, suggesting that carotenoid-based plumage coloration is environmentally determined. However signaling functions ascribed to plumage imply that plumage coloration is the outcome of an evolutionary process based on genetic variation. By means of a cross-fostering design we show significant effects of both a common rearing environment and the brood from which a nestling originally came from (common origin) on the plumage coloration of nestling great tits (Parus major). This demonstration of origin-related variation in carotenoid-based plumage coloration suggests that the observed variation of the trait has a partial genetic basis. Consistent with environmental determination of this trait, we also found a significant positive correlation between the color saturation of nestlings and their foster-father's plumage. There was no significant correlation between nestling plumage coloration and the food quantity provided to the nestlings by the male, the female, or both parents. This suggests that the nestling-foster father correlation arises by the carotenoid quantity ingested rather than the food quantity per se. No significant nestling-true father correlation was found, which suggests that nestling plumage coloration did not indirectly evolve due to sexual selection. Consistent with this result there was no significant correlation between the nestling's plumage color and its coloration as a breeding adult the following year, suggesting that nestling plumage color is a different trait than the first year plumage. PMID- 12643575 TI - Accumulation of Dobzhansky-Muller incompatibilities within a spatially structured population. AB - A simple, deterministic analysis predicts that accumulation of Dobzhansky-Muller incompatibilities by a spatially structured population strongly depends on the number of negative interactions of an allele. If an allele can be incompatible with alleles at only one locus, incompatibilities accumulate linearly with time. In contrast, if an allele can participate in multiple pairwise incompatibilities with alleles at different loci, the expected number of incompatibilities eventually increases quadratically. PMID- 12643576 TI - Socially mediated speciation. AB - We employ a simple model to show that social selection can lead to prezygotic reproductive isolation. The evolution of social discrimination causes the congealing of phenotypically similar individuals into different, spatially distinct tribes. However, tribal formation is only obtained for certain types of social behavior: altruistic and selfish acts can produce tribes, whereas spiteful and mutualistic behaviors never do. Moreover, reduced hybrid fitness at tribal borders leads to the selection of mating preferences, which then spread to the core areas of the respective tribes. Unlike models of resource competition, our model generates reproductive isolation in an ecologically homogeneous environment. We elaborate on how altruistic acts can lead to reproductive isolation, but also predict that certain types of competition can lead to the speciation effect. Our theory provides a framework for how individual-level interactions mold lineage diversification, with parapatric speciation as a possible end product. PMID- 12643577 TI - Worldwide distribution of transposable element copy number in natural populations of Drosophila simulans. AB - Transposable elements (TEs), which promote various kinds of mutations, constitute a large fraction of the genome. How they invade natural populations and species is therefore of fundamental importance for understanding the dynamics of genetic diversity and genome composition. On the basis of 85 samples of natural populations of Drosophila simulans, we report the distributions of the genome insertion site numbers of nine TEs that were chosen because they have a low average number of sites. Most populations were found to have 0-3 insertion sites, but some of them had a significantly higher number of sites for a given TE. The populations located in regions outside Africa had the highest number of sites for all elements except HMS Beagle and Coral, suggesting a recent increase in the activity of some TEs associated with the colonization patterns of Drosophila simulans. The element Tirant had a very distinctive pattern of distribution: it was identified mainly in populations from East Africa and some islands in the Indian Ocean, and its insertion site number was low in all these populations. The data suggest that the genome of the entire species of Drosophila simulans may be being invaded by TEs from populations in which they are present in high copy number. PMID- 12643578 TI - One host shift leads to another? Evidence of host-race formation in a predaceous gall-boring beetle. AB - We show that a predator, the tumbling flower beetle Mordellistena convicta (Coleoptera: Mordellidae), has formed host races in response to a host-plant shift and subsequent host-race formation by its prey, the gall-inducing fly Eurosta solidaginis (Diptera: Tephritidae). This fly has formed two host races, one that induces stem galls on the ancestral host plant, Solidago altissima (Compositae), and another that induces stem galls on the closely related S. gigantea. We found that subpopulations of M. convicta that attack E. solidaginis galls on the different host plants have significantly different emergence times and, although slight, these allochronic differences are consistent across a range of temperatures. More importantly, we found that beetles assortatively mate according to their natal host plants, and female M. convicta preferentially attack and/or their offspring have higher survival in galls on natal host plants. Our data suggest that subpopulations of M. convicta that attack E. solidaginis galls on S. altissima and S. gigantea have formed host races. This is one of the first studies to demonstrate that a host shift and subsequent host-race formation by an herbivorous insect may have resulted in subsequent diversification by one of its natural enemies. PMID- 12643579 TI - Linear models for assessing mechanisms of sperm competition: the trouble with transformations. AB - Although sperm competition is a pervasive selective force shaping the reproductive tactics of males, the mechanisms underlying different patterns of sperm precedence remain obscure. Parker et al. (1990) developed a series of linear models designed to identify two of the more basic mechanisms: sperm lotteries and sperm displacement; the models can be tested experimentally by manipulating the relative numbers of sperm transferred by rival males and determining the paternity of offspring. Here we show that tests of the model derived for sperm lotteries can result in misleading inferences about the underlying mechanism of sperm precedence because the required inverse transformations may lead to a violation of fundamental assumptions of linear regression. We show that this problem can be remedied by reformulating the model using the actual numbers of offspring sired by each male, and log-transforming both sides of the resultant equation. Reassessment of data from a previous study (Sakaluk and Eggert 1996) using the corrected version of the model revealed that we should not have excluded a simple sperm lottery as a possible mechanism of sperm competition in decorated crickets, Gryllodes sigillatus. PMID- 12643580 TI - Population on the verge of a mutational meltdown? Fitness costs of genetic load for an amphibian in the wild. AB - The fitness costs of high genetic load in wild populations have rarely been assessed under natural conditions. Such costs are expected to be greatest in small, bottlenecked populations, including those occurring near range edges. Britain is at the northwesterly range limit of the natterjack toad Bufo calamita. We compared fitness attributes in two populations of this amphibian with very different recent histories. Key larval fitness attributes in B. calamita, notably growth rate and metamorph production, were substantially higher in the large outbreeding population (Ainsdale) than in the small and isolated one (Saltfleetby). These differences were manifest under seminatural conditions, when larvae were reared in mesh cages within breeding ponds at the site of the small population, and were exacerbated by high stress treatments. The results indicate that genetic load effects can be sufficiently severe enough to predispose extinction over relatively short time frames, as predicted by extinction vortex models. PMID- 12643581 TI - Stochasticity overrules the "three-times rule": genetic drift, genetic draft, and coalescence times for nuclear loci versus mitochondrial DNA. AB - Palumbi et al. (2001) proposed a "three-times rule" that uses mitochondrial DNA (mtDNA) sequences to predict probabilities of monophyly for nuclear loci (i.e., whether the alleles within a taxon coalesce with one another before they coalesce with alleles from a sister taxon). They use neutral coalescent theory to infer these probabilities from the ratio of interspecific divergence to intraspecific variation of mtDNA. We show that the estimated probabilities have very wide confidence intervals because of the inherent stochasticity of the mtDNA coalescent process. Under neutrality, the true probability of monophyly can be much higher, or much lower, than predicted by the three-times rule. We also review recent empirical and theoretical studies that refute neutrality-based predictions concerning mtDNA variation and divergence. We conclude that the three times rule is neither a useful test for neutral molecular evolution nor a reliable guide to genealogical species. PMID- 12643582 TI - Quantitative genetics of geometric shape: heritability and the pitfalls of the univariate approach. PMID- 12643583 TI - Shape distances in general linear models: are they really at odds with the goals of morphometrics? A reply to Klingenberg. PMID- 12643584 TI - Synthesis of certain substituted quinoxalines as antimicrobial agents (Part II). AB - Several fused triazolo and ditriazoloquinoxaline derivatives such as 1-aryl-4 chloro-[1,2,4]triazolo[4,3-a]quinoxalines (3a-d), 4-alkoxy[1,2,4]triazolo[4,3 a]quinoxalines (4a,b), 4-substituted-amino-[1,2,4] triazolo[4,3-a]quinoxalines (5a-h), 1-(aryl)-[1,2,4]triazolo[4,3-a]quinoxalin-4(5H)-thione (6), 4 (arylidenehydrazino)1-phenyl-[1,2,4]triazolo[4,3-a]quinoxalines (10a-e) and [1,2,4]ditriazolo[4,3-a:3',4'-c]quinoxaline derivatives (11-13) have been synthesized and some of these derivatives were evaluated for antimicrobial and antifungal activity in vitro. It was found that compounds 3a and 9b possess potent antibacterial activity compared to the standard tetracycline. PMID- 12643585 TI - Achiral and chiral determination of benzophenanthridine alkaloids from methanol extracts of Hylomecon species by high performance liquid chromatography. AB - A high performance liquid chromatographic (HPLC) method was developed for the qualitative and quantitative determination of benzophenanthridine alkaloids from the methanol extracts of Hylomecon hylomeconoides and H. vernale (Papaveraceae). Achiral and chiral methods were adapted for the separation of 6 methoxydihydrosanguinarine (1), 6-acetonyldihydrosanguinarine (2) and dihydrosanguinarine (3). The achiral reversed phase HPLC method made it possible the simultaneous separation and determination of 1, 2 and 3 within 20 min on ODS column using acetonitrile-phosphate buffer (50 mM, pH 7.0) (50:50, v/v). The separation and determination of 1 and 2 enantiomers was available using chiral columns. The same amount of (+) and (-)-enantiomers of 1 was found from the methanol extract of specimen, indicated that 1 could be the artifact produced by the reaction of sanguinarine with methanol. H. hylomeconoides showed higher level of 1 and 3 in compared with H. vernale, especially in root samples permitting the possibility of chemical discrimination between two species. PMID- 12643586 TI - Determination of terbutaline enantiomers in human urine by capillary electrophoresis using hydroxypropyl-beta-cyclodextrin as a chiral selector. AB - A method for the determination of terbutaline enantiomers in human urine by capillary electrophoresis has been developed. Optimum resolution was achieved using 50 mM phosphate buffer, pH 2.5, containing 15 mM of hydroxypropyl-beta cyclodextrin as a chiral selector. Urine samples were prepared by solid-phase extraction with Sep-pak silica, followed by CE. The assay was linear between 2 250 ng/mL (R = 0.9998 for (S)-(+)-terbutaline and R = 0.9999 for (R)-( )terbutaline) and detection limit was 0.8 ng/mL. The intra-day variation ranged between 6.3 and 14.5% in relation to the measured concentration and the inter-day variation was 8.2-20.1%. It has been applied to the determination of (S) (+)terbutaline and (R)-(-)-terbutaline in urine from healthy volunteer dosed with racemic terbutaline sulfate. PMID- 12643588 TI - A polyacetylene and flavonoids from Cirsium rhinoceros. AB - Cirsium rhinoceros Nakai (Compositae) is a herbaceous perennial native to Korea, whole plant of which has been used in folklore medicine. C. rhinoceros was extracted by a standard extraction procedure. Its n-hexane, CHCl3 and n-BuOH extracts were fractionated by column chromatography to provide a polyacetylene, a coumarin and five flavonoids. Ciryneol C, scopoletin, acacetin, cirsimarin were isolated for the first time from this plant. PMID- 12643587 TI - Bioactive prenylated flavonoids from the stem bark of Artocarpus kemando. AB - Four known prenylated flavonoids, artonins E (1) and O (2), artobiloxanthone (3), and cycloartobiloxanthone (4), were isolated from the stem bark of Artocarpus kemando by bioassay-guided fractionation using the DNA strand-scission and the KB cytotoxicity assays as monitors. Compounds 1 and 3 exhibited strong DNA strand scission activity, and all four compounds were found to be cytotoxic. PMID- 12643589 TI - Cerebrosides and terpene glycosides from the root of Aster scaber. AB - Three cerebrosides 2, 3, and 5 and two terpene glycosides 1 and 4 have been isolated from the methanol extract of the root of Aster scaber. Their structures were determined as 3-O-beta-D-glucuronopyranosyl-oleanolic acid methyl ester (1), (2S, 3S, 4R, 2'R, 8Z, 15'Z)-N-2'-hydroxy-15'-tetracosenoyl-1-O-beta-D glucopyranosyl-4-hydroxy-8-sphingenine (2), (2S, 3S, 4R, 8Z)-N-octadecanoyl-1-O beta-D-glucopyranosyl-4-hydroxy-8-sphingenine (3), 1alpha-hydroxy-6beta-O-beta-D glucosyl-eudesm-3-ene (4), and (2S, 3S, 4R, 2'R, 8Z)-N-2'-hydroxy-hexadecanoyl-1 O-beta-D-glucopyranosyl-4-hydroxy-8-sphingenine (5) on the basis of spectroscopic methods. PMID- 12643590 TI - Cerebrosides from Longan Arillus. AB - From the pulp of Euphoria longana (Longan Arillus), three cerebroside molecular species have been isolated. Six known cerebrosides, soyacerebrosides I and II, 1 O-beta-D-glucopyranosyl-(2S,3R,4E,8E)-2-(2'-lignoceroylamino)-4,8-octadecadiene 1,3-diol (longan cerebroside I) and its 8Z isomer (longan cerebroside II), momor cerebroside I, and phytolacca cerebroside, were identified as major components of these cerebroside molecular species. All the cerebrosides were shown to be a mixture of geometrical isomers (8E and 8Z) of sphingosine-type or phytosphingosine-type glucocerebrosides possessing 2-hydroxy fatty acids. The structures of these cerebrosides have been determined on the basis of chemical and spectroscopic evidence. PMID- 12643591 TI - In vivo anti-nociceptive and anti-inflammatory effect of the two triterpenes, ursolic acid and 23-hydroxyursolic acid, from Cussonia bancoensis. AB - Triterpenoids, ursolic acid (1) and 23-hydroxyursolic acid (2) were obtained from the hydrolysis of BuOH fraction of Cussonia bancoensis extract to test antinociceptive and anti-inflammatory effect of C. bancoensis (Araliaceae). Compound 1 and 2 exhibited anti-nociceptive effects, which were determined by acetic acid-induced writhing test and hot plate test. The effect of 2 was much more potent in acetic acid-induced writhing test than in hot plate test. Compound 1 and 2 significantly inhibited 1%-carrageenan-induced edema in the rat. These results suggest that the two triterpenes, ursolic acid and 23-hydroxyursolic acid, are responsible for the anti-nociceptive and anti-inflammatory effect of C. bancoesnsis. PMID- 12643592 TI - A DNA strand-nicking principle of a higher plant, Caesalpinia sappan. AB - To find anticancer agents from higher plants, DNA strand-scission assay method was employed for bioassay-guided fractionation as well as for screening the crude extracts. During the screening, an ethyl acetate extracts of the heartwood of Caesalpinia sappan L. (Leguminosae) exhibited potent DNA strand-scission activity. Therefore, the ethyl acetate extracts of the dried heartwood of C. sappan was subjected to the bioassay-guided fractionation, which led to the isolation of a known compound, brazilin (1) as the active constituent. In addition, caesalpine J (2) was also isolated as an inactive constituent. PMID- 12643593 TI - Inhibitory effects of luteolin isolated from Ixeris sonchifolia Hance on the proliferation of HepG2 human hepatocellular carcinoma cells. AB - We investigated the anti-proliferative effects of luteolin and apigenin, isolated from Ixeris sonchifolia Hance, on HepG2 human hepatocellular carcinoma cells. In MTT assay luteolin showed more efficient anti-proliferative effects on cells than apigenin did. According to propidium iodide staining and flow cytometry studies, we postulated that these effects might be a result of cell cyde arrest. Hence we examined the changes of protein expressions related to cell cycle arrest. Western blotting data demonstrated that the down-regulated expression of CDK4 was correlated to the increase of p53 and CDK inhibitor p21(WAF1/CIP1) protein. These data suggest that luteolin may have potential as an anti-cancer agent. PMID- 12643594 TI - Apoptosis induction of Persicae Semen extract in human promyelocytic leukemia (HL 60) cells. AB - The major ingredient of Persicae Semen is a cynogenic compound, amygdalin (D mandelonitrile-beta-gentiobioside). Controversial results on the anticancer activity of amygdalin were reported due to its conversion to its inactive isomer, neoamygdalin. In order to inhibit the epimerization of amygdalin, we used newly developed simple acid boiling method in preparation of Persicae Semen extract. HPLC analysis revealed most of amygdalin in Persicae Semen extract was active D form. Persicae Semen extract was used to analyze its effect on cell proliferation and induction of apoptosis in human promyelocytic leukemia (HL-60) cells. Persicae Semen extract was cytotoxic to HL-60 cells with IC50 of 6.4 mg/mL in the presence of 250 nM of beta-glucosidase. The antiproliferative effects of Persicae Semen extract appear to be attributable to its induction of apoptotic cell death, as Persicae Semen extract induced nuclear morphology changes and internucleosomal DNA fragmentation. PMID- 12643595 TI - Physicochemical characterization and in vivo evaluation of thermosensitive diclofenac liquid suppository. AB - Liquid suppository systems composed of poloxamers and bioadhesive polymers were easy to administer to the anus and mucoadhesive to the rectal tissues without leakage after the dose. However, a liquid suppository containing diclofenac sodium could not be developed using bioadhesive polymers, since the drug was precipitated in this preparation. To develop a liquid suppository system using sodium chloride instead of bioadhesive polymers, the physicochemical properties such as gelation temperature, gel strength and bioadhesive force of various formulations composed of diclofenac sodium, poloxamers and sodium chloride were investigated. Furthermore, the pharmacokinetic study of diclofenac sodium delivered by the liquid suppository was performed. Diclofenac sodium significantly increased the gelation temperature and weakened the gel strength and bioadhesive force, while sodium chloride did the opposite. The liquid suppositories with less than 1.0% of sodium chloride, in which the drug was not precipitated, were inserted into the rectum without difficulty and leakage. Furthermore, liquid suppository gave significantly higher initial plasma concentrations and faster Tmax of diclofenac sodium than did solid suppository, indicating that drug from liquid suppository could be absorbed faster than that from solid one in rats. Our results suggested that a thermosensitive liquid suppository system with sodium chloride and poloxamers was a more physically stable, convenient and effective rectal dosage form for diclofenac sodium. PMID- 12643596 TI - DA-7911, 188Rhenium-tin colloid, as a new therapeutic agent of rheumatoid arthritis. AB - Radiation synovectomy is one of the most useful methods for treating patients with refractory synovitis because of its convenience, long-term effects, repeatability and the avoidance of surgery. In this study, we investigated the toxicity, stability and biodistribution of a rhenium-188 (188Re)-tin colloid to evaluate its suitability as a synovectomy agent. Twenty four hours after injecting the 188Re-tin colloids (74 KBq/0.1 mL) into the tail vein of ICR mice, most of the 188Retin colloidal particles was found in the lungs. In addition, there were no particle size changes at either room temperature or at 37 degrees C after injecting the 188Re-tin colloids in human plasma and synovial fluid. In vitro stability tests showed that the 188Re-tin colloid remained in a colloidal form without a critical size variation over a 2-day period. We investigated the leakage of 188Retin colloids from the intraarticular injection site with gamma counting in New Zealand white rabbits. The 188Re-tin colloids (55.5 MBq/0.15 mL) were injected at the cavum articular and the mean retention percentage of the 188Re-tin colloid was 98.7% for 1 day at the injection site, which suggests that there was neither change in the particle size nor leakage at the injection sites. In the biodistribution study with the SD rats, the liver showed the highest radioactivity (0.0427% ID/organ) except for the injected knees (99.49%). In the SD rats, mild toxicities including the skin or a synovium inflammation were observed as a result of a radioactivity of 15 mCi/kg at the intraarticular injection site. However, there was no systemic toxicity. In the Ovalbumin (OVA) induced arthritic rabbits, the 188Re-tin colloid improved the macroscopic, the histological score and reduced the knee joint diameter when compared to the arthritic control. In conclusion, a 188Re-tin-colloid is considered as a strong candidate for radiation synovectomy with a superior efficacy and safety. PMID- 12643597 TI - A polymeric micellar carrier for the solubilization of biphenyl dimethyl dicarboxylate. AB - A polymeric micelle drug delivery system was developed to enhance the solubility of poorly-water soluble drug, biphenyl dimethyl dicarboxylate, DDB. The block copolymers consisting of poly(D,L-lactide) (PLA) as the hydrophobic segment and methoxy poly(ethylene glycol) (mPEG) as the hydrophilic segment were synthesized and characterized by NMR, DSC and MALDI-TOF mass spectroscopy. The size of the polymeric micelles measured by dynamic light scattering showed a narrow monodisperse size distribution with the average diameter less than 50 nm. The MW of mPEG-PLA, 3000 (MW of mPEG, 2 K; MW of PLA, 1 K), and the presence of hydrophilic and hydrophobic segments on the polymeric micelles were confirmed by MALDI-TOF mass spectroscopy and NMR, respectively. Polymeric micelle solutions of DDB were prepared by three different methods, i.e. the matrix method, emulsion method and dialy-sis method. In the matrix method, DDB solubility was reached to 13.29 mg/mL. The mPEG-PLA 2K-1 K micelle system was compared with the poloxamer 407 micelle system for their critical micelle concentration, micelle size, solubilizing capacity, stability in dilution and physical state. DDB loaded polymeric micelles prepared by the matrix method showed a significantly increased aqueous solubility (>5000 fold over intrinsic solubility) and were found to be superior to the poloxamer 407 micelles as a drug carrier. PMID- 12643599 TI - Electrical pacing accelerates intestinal transit slowed by fat-induced ileal brake. AB - Previous studies on intestinal electrical stimulation were aimed at achieving a delay in intestinal transit. The aim of this study was to test the hypothesis that the physiological slowing of intestinal transit by the fat-induced ileal brake might be accelerated with intestinal electrical stimulation. This study was performed in five dogs prepared with eight serosal electrodes in the jejunum and two chronic intestinal fistulas located in the duodenum and at the midpoint of the small intestine. To slow transit by triggering the ileal brake, oleate was delivered into the distal half of the gut while buffer was perfused into the proximal half of the gut. Intestinal transit between the fistulas was measured by the recovery of [99mTc]DTPA. To test for the effect of pacing, transit was compared with vs. without continuous forward electrical stimulation (frequency: 24 cycles/min; pulse duration: 50 msec, pulse amplitude 1-3 mA). Electrical stimulation completely entrained intestinal pacesetter potentials as measured fromthe seven recording electrodes distal to the pacing electrode. A substantial and significant increase in intestinal transit was observed with intestinal electrical stimulation with the percentage of marker recovery increasing from 19.2 +/- 9.3% to 84.6 +/- 11.3% (P < 0.01). In conclusion, intestinal electrical stimulation accelerates intestinal transit slowed by ileal brake. PMID- 12643598 TI - Racecadotril versus loperamide: antidiarrheal research revisited. AB - Racecadotril is an enkephalinase inhibitor, presented as a purely antisecretory agent with advantages over the opiate-receptor agonist loperamide in the treatment of diarrhea. A critical review of the literature and the models used was performed. Although pretreatment with high doses of racecadotril reduced cholera toxin-induced secretion and although clinical efficacy was demonstrated in young infants--a population characterized by 10-fold higher plasma enkephalin concentrations compared with adults, the analysis calls into question the peripheral antisecretory selectivity and relative clinical efficacy. Conversely, loperamide can be proposed as an antisecretory agent at therapeutic concentrations. Its efficacy is well established in acute and chronic diarrhea. Current experimental and clinical comparative studies of both drugs have problems with regard to the selection of the doses, the validity of models, and/or the trial design. The conclusion is that more research is needed before reliable conclusions can be drawn on the place of racecadotril in diarrhea treatment. PMID- 12643600 TI - Role of NF-kappaB and AP-1 on Helicobater pylori-induced IL-8 expression in AGS cells. AB - Oxygen radicals are important regulators in Helicobacter pylori-induced gastric ulceration and carcinogenesis. IL-8 may be regulated by oxidant-sensitive transcription factors, NF-kappaB, and AP-1. The present study aims to investigate whether H. pylori-induced IL-8 expression is regulated by NF-kappaB and AP-1 in gastric epithelial AGS cells and whether this transcriptional regulation of IL-8 is inhibited by N-acetylcysteine (NAC). As a result, H. pylori induced the expression of mRNA and protein for IL-8 via activation of NF-kappaB and AP-1. NF kappaB activation accompanied by a decrease in I-kappaBalpha and activated AP-1 complex was a c-jun/c-fos heterodimer in H. pylori-infected AGS cells. NAC inhibited H. pylori-induced activation of transcription factors and IL-8 expression in AGS cells. In conclusion, oxygen radicals induce the activation of NF-kappaB and AP-1 and IL-8 expression. Antioxidants such as NAC might be useful anti-inflammatory agents by inhibiting activation of transcription factors and decreasing IL-8 production in H. pylori-induced gastric inflammation. PMID- 12643601 TI - Expression of survivin and caspase-3 in gastric cancer. AB - Gastric cancer is one of the most common malignant tumors of the gastrointestinal tract. However, the molecular pathways involved in the regulation of gastric carcinogenesis are not completely elucidated. In the last decade, basic cancer research has been focused on the deregulation of apoptosis as a central event in the process of carcinogenesis. Caspase-3 and survivin are regulators of apoptosis and have been implicated in the development of gastric cancer. The aim of the present study was to compare the expression of mRNA and protein for survivin and caspase-3 in the gastric cancer and in the cancer margin with that in normal human gastric mucosa. Fifteen patients with advanced gastric cancer (all H. pylori-positive) and 15 matched control subjects with normal gastric mucosa were included in this study. The biospy specimens for histology and for molecular analyses were taken from gastric tumor, tumor surrounding gastric mucosa and in normal patients from the mucosa of antrum and corpus. Survivin mRNA expression was very weak, but detectable, in the normal gastric mucosa. However, at the protein level, no expression for survivin was detected in the normal gastric mucosa. In the biopsy specimens from tumor and surrounding gastric mucosa, a significant increase in survivin mRNA and protein expression was observed. The expression of survivin was higher in the tumor than in the tumor margin. The mRNA and protein expression of caspase-3 was detected in the gastric mucosa of normal subjects. In gastric cancer only the expression of procaspase-3 was observed, while the expression of active caspase-3 was completely undetectable. In the gastric mucosa surrounding gastric cancer, no gene and protein expression for caspase-3 was detected. We conclude that the changes in the level of caspase-3 and survivin play an important role in the transformation from normal gastric mucosa to gastric career. PMID- 12643602 TI - Laminin-5 gamma2 chain expression as a possible determinant of tumor aggressiveness in T1 colorectal carcinomas. AB - This study was undertaken to clarify the associations between laminin-5 y2 chain expression, and tumor budding and lymph node metastasis or local recurrence (locoregional failure) of 50 T1 colorectal carcinomas immunohistochemically. Fifteen (30%) of 50 lesions were positive for laminin-5 gamma2 chain expression. By univariate analysis, less histological differentiation (P = 0.02), nonpolypoid growth pattern (P = 0.03) and tumorbudding (P < 0.001) were associated with laminin-5 gamma2 chain positivity. By multivariate analysis, tumor budding alone was significantly associated with laminin-5 gamma2 chain positivity (P = 0.006), and correlation between nonpolypoid growth pattern and laminin-5 gamma2 chain positivity lost its significance (P = 0.09). Tumor budding (P = 0.004) and laminin-5 gamma2 chain expression (P = 0.001) were significantly associated with locoregional failure. Laminin-5 gamma2 chain expression may contribute to the formation of budding tumor cells at the invasive front, and immunostaining of this adhesion molecule may be useful in identifying high-risk patients for locoregional failure in T1 colorectal carcinomas. PMID- 12643604 TI - Pedunculated gastrointestinal stromal tumor presenting as bowel volvulus: usefulness of color Doppler ultrasonography for strangulated ileus. PMID- 12643605 TI - Rapid four-month growth of an early-stage adenocarcinoma of the colon with neuroendocrine characteristics. PMID- 12643603 TI - In vivo assessment of angioarchitecture and microcirculation in experimental liver cancer: a new model in rats. AB - The rising incidence of unresectable hepatocellular malignancies remains a therapeutic challenge. Little is known about the mechanisms of angiogenesis and immunological aspects of liver tumor vessels. The aim of this study was to investigate hepatic and tumor microcirculation and leukocyte-endothelium interaction by means of intravital microscopy in a rat model of hepatocellular carcinoma. Off-line analysis showed that the angioarchitecture as well as blood flow velocity in liver cancer is heterogeneous. The leukocyte-endothelium interaction is significantly reduced compared to normal liver tissue. The data suggest that the main mechanism is a reduced expression of adhesion molecules demonstrating an effective immune escape mechanism of this tumor. The model represents a useful experimental tool to explore angiogenesis inhibition or immunological therapeutic strategies in experimental liver cancer. PMID- 12643606 TI - Schatzki's rings do not protect against acid reflux and may decrease esophageal acid clearance. AB - Schatzki's rings (SR) are a common cause of intermittent solid food dysphagia, but their etiology is unclear. Many believe they are related to acid reflux, hypothesizing that the rings act as a protective barrier against further reflux. The purpose of this study was to determine whether dilation of SR affected the degree of acid reflux. Twenty patients participated in the study. All patients underwent esophageal manometry and 24-hr pH monitoring off all acid inhibitory medications before and two weeks after esophageal dilation. No significant differences were noted in any of the reflux parameters measured before and after dilation. However, there was a trend toward reduction in symptom score in all patients, a decrease in Johnson-DeMeester score, and a decrease in supine reflux time in patients with thick SR after dilation. There was no correlation between ring diameter and the presence or absence of reflux. In conclusion, Schatzki's rings do not prevent esophageal reflux, and they may act to decrease esophageal acid clearance, especially in the supine position, thereby increasing esophageal acid exposure. PMID- 12643607 TI - Relationship of sliding hiatus hernia to gastroesophageal reflux disease: a possible role for Helicobacter pylori infection? AB - Sliding hiatal hernia is a common endoscopic finding with a prevalence that increases with the age of patients. Although nearly all patients with GERD have HH, only a minority of patients with hernia reports reflux symptoms. Our hypothesis is that H. pylori infection may be responsible for the high number of asymptomatic hernias. After exclusion of patients with peptic ulcer, 507 patients with an endoscopic diagnosis of hernia were considered. Patients were divided into three groups: A, < or = 45 years, 141 patients; B, 46-60 years, 144 patients; and C, > or = 61 years, 222 patients. Presence of reflux symptoms (questionnaire) and esophagitis, H. pylori status, and gastric histology were recorded. The prevalence of hernia in the total series was 11% in group A, 23% in B, and 38% in C. Aging was associated with a significant increase in H. pylori prevalence and corpus gastritis scores, and a parallel decrease of GERD symptom prevalence, which was 66.6% in group A, 52.1% in B, and 46.8% in C (P < 0.01). Taking the three groups together, prevalence of H. pylori infection was higher in patients without GERD than with GERD (66.4 vs 57.3%, P < 0.05), and higher in patients with nonerosive GERD than erosive GERD (62.8 vs 48.6%, P = 0.02); corpus gastritis scores were significantly higher in patients without GERD than those with GERD and in those with nonerosive than erosive GERD. In conclusion, H. pylori infection protects against development of GERD in subjects with hiatus hernia. This effect is significantly more evident in the elderly where, in spite of the high prevalence of hernia, only a small number of individuals develop GERD. The development of a corpus-predominant gastritis is probably responsible for this effect. PMID- 12643608 TI - Role of gastric mucosal and gastric juice cytokine concentrations in development of bisphosphonate damage to gastric mucosa. AB - Previous studies have shown that the bisphosphonates (BP) vary in their damaging effect on the gastric mucosa, and endoscopy scores (erosions or erosions plus ulcers) after 1 and 2 weeks use of BP were significantly lower in H. pylori positive versus -negative subjects. The mechanism of this damaging effect of BP and the interaction with H. pylori is unknown. As part of a separately reported study of the incidence of gastric damage after 2 weeks of treatment of healthy female postmenopausal volunteers with risedronate (5 mg/day) or alendronate (10 mg/day), gastric aspirates were taken at the time of the baseline esophagogastroduodenoscopy (EGD), and again at 1 and 2 weeks after daily intake of a BP At the time of the third EGD, when the volunteers had been on risedronate or alendronate for 2 weeks, antral biopsies were taken from normal-appearing mucosa. Gastric juice and antral biopsies were assessed for their concentration of the cytokines interleukin-la (IL-1alpha), IL-8, IL-13, and epidermal growth factor (EGF). H. pylori, the use of BP, and development of gastric mucosal lesions had no effect on gastric mucosal concentrations of IL-1alpha, IL-13, or EGF. In contrast, the concentration of IL-8 in antral mucosal biopsies of volunteers given BP for 2 weeks was higher in the presence than in the absence of an H. pylori infection and was increased further in those who develop lesions associated with the use of BP. There was no correlation between gastric mucosal and gastric juice concentrations of IL-8. Gastric juice concentrations of IL-8 and EGF were not affected by H. pylori status, the use of BP, or the development of lesions. However, gastric juice concentrations of IL-1alpha were numerically lower in those who were negative for H. pylori with no mucosal lesions (Hp-L-), intermediate in those who were H. pylori-negative with lesions (Hp-L+), and highest in those who were positive for H. pylori and had lesions (Hp+L+). The gastric juice concentration of IL-13 was threefold higher in the absence than in the presence of H. pylori, and the relative abundance of IL-13 was: Hp-L- >Hp-L+ >Hp+L(-1) >Hp+L+. The prostaglandin E2 concentration in gastric antral biopsies was similar in the four groups and was unchanged with the in vitro biopsy incubation with celecoxib. We speculate that the higher gastric endoscopy scores observed with the use of BP in H. pylori negative as compared with H. pylori positive individuals is due to their lower mucosal concentration of IL-8 as well as the lower gastric juice concentration of IL-1alpha and higher concentration of IL-13. PMID- 12643609 TI - Impact of long-term ranitidine and pantoprazole on accuracy of [13C]urea breath test. AB - No previous study has analyzed the impact of long-term antisecretory drugs on the precision of [13C]urea breath test (UBT). We assessed the rate of UBT conversion from positive to negative results during 60-day therapy with standard doses of ranitidine and pantoprazole. For this purpose, we recruited 60 dyspeptic patients with H. pylori infection ascertained on the basis of the concomitant results of CLO-test, histology, and UBT. Our patients were randomly assigned to receive ranitidine 300 mg at night or pantoprazole 40 mg in the morning for 60 days. UBT was performed at baseline and on days 14, 30, and 60, while patients were still taking antisecretory drugs. Patients with false-negative UBT on day 60 repeated the test every 3 days until conversion. After overnight fasting, duplicate breath test samples were taken from each patient before and 30 min after ingestion of 75 mg [13C]urea dissolved in 150 ml of 0.033 mol/liter citric acid. Four patients dropped out of the study. Both drugs induced similar false-negative UBTs on day 14 of dosing (P = 0.5). Afterwards, the three false-negative UBTs in the ranitidine group again became positive during therapy and particularly on day 30 of dosing. Of the four false-negative UBTs in the pantoprazole group at day 60, one became positive after 3 and three after 9 days of therapy cessation. Our findings show that the long-term use of ranitidine and pantoprazole at standard doses has different effects on the results of UBT. In the pantoprazole group patients again became positive within 3-9 days after stopping 60-day therapy, whereas in the ranitidine group patients reverted to positive on day 30 of dosing while they were still on treatment and this was likely due to development of tolerance. Therefore, patients taking pantoprazole need at least a 10-day withdrawal before UBT testing, while those taking ranitidine for at least 30 days can undergo UBT without the necessity of a wash-out period. PMID- 12643611 TI - Mechanisms of transforming growth factor-alpha (TGF-alpha) induced gastroprotection against ethanol in the rat: roles of sensory neurons, sensory neuropeptides, and prostaglandins. AB - The mechanisms by which transforming growth factor-a (TGP-alpha) protects the stomach against mucosal injury are incompletely understood. The aim of this study was to examine the roles of sensory neurons, sensory neuropeptides and prostaglandins in TGFalpha gastroprotection against ethanol. Fasted rats received TGF-alpha (50 microg/kg, intraperitoneally) prior to orogastric ethanol (75% v/v, 1 ml). Gastric injury was quantitated 30 min after ethanol. Involvement of sensory neurons and the sensory neuropeptides, calcitonin gene-related peptide (CGRP) and substance P (SP), were examined by capsaicin deafferentation and specific receptor antagonist infusion, respectively. Indomethacin (10 mg, intragastrically) was used to determine the role of prostaglandins in TGF-alpha mediated gastroprotection. TGF-alpha significantly diminished ethanol-induced gastric lesion area to 5.7 +/- 0.8 mm2 vs 41.1 +/- 5.2 mm2 (p < 0.001). Sensory denervation and CGRP-receptor blockade abolished the TGF-alpha protective effect. In contrast, SP antagonist and indomethacin did not alter TGF-alpha gastroprotection. In conclusion, TGF-alpha-mediated gastroprotection involves sensory neuron activation and CGRP release and this protective effect did not involve substance P or prostaglandin generation. PMID- 12643610 TI - Significant enhancement of gastric mucin content after rabeprazole administration: its potential clinical significance in acid-related disorders. AB - Rabeprazole is the only proton pump inhibitor that enhances the content of gastric mucin in experimental animals. We have studied, therefore, the effect of rabeprazole on the content of gastric mucin, mucus, and its viscosity in 21 asymptomatic volunteers in a double-blind study. The mucus content during rabeprazole administration significantly increased both in pentagastrin stimulated (3.36 +/- 0.39 vs 1.50 +/- 0.32 mg/ml, P < 0.001) and basal (3.31 +/- 0.38 vs 2.28 +/- 0.36 mg/ml, P < 0.01) conditions. The content of mucin during rabeprazole was 2.6-fold (0.96 +/- 0.08 vs 0.36 +/- 0.06 mg/ml, P < 0.0001) and 41% (0.82 +/- 0.09 vs 0.58 +/- 0.09 mg/ml, P < 0.05) higher in stimulated and basal conditions, respectively. The viscosity of gastric juice during rabeprazole administration was also significantly higher both in stimulated (P < 0.01) and basal (P < 0.05) conditions. In conclusion, the unique pharmacological property of rabeprazole, significantly augmenting production of gastric mucus and mucin, may translate to additional clinical benefits in protecting the upper alimentary tract mucosa during the acid-related challenge. PMID- 12643612 TI - Pharmacological evidence for beta3 adrenoceptors in the control of rat gastric acid secretion. AB - The effect of the beta3-adrenoceptor agonist BRL37344 on gastric acid secretion evoked by different secretory stimuli was investigated in anaesthetized rats with lumen-perfused stomachs in comparison with the beta2-adrenoceptor agonist clenbuterol. Intravenous injections of BRL37344 (1-10 micromol/kg) and clenbuterol (0.01-1 micromol/kg) dose-dependently reduced 2-deoxy-D-glucose induced acid secretion, with BRL37344 about forty times less potent than clenbuterol. BRL37344 (0.1-3 micromol/kg) inhibited pentagastrin-induced acid output, whereas clenbuterol was effective only at high doses (10-100 micromol/kg). The inhibitory effect of BRL37344 on pentagastrin-induced acid secretion was not modified by the nonselective beta-adrenoceptor antagonist propranolol, but it was prevented by bupranolol, a beta3-adrenoceptor antagonist. Furthermore, neither BRL37344 (10 micromol/kg) nor clenbuterol (100 micromol/kg) modified the acid secretion induced by histamine. These data suggest that beta3 adrenoceptors have an inhibitory role in the control of rat gastric acid secretion induced by indirect stimuli. PMID- 12643614 TI - High prevalence of non-organ-specific autoantibodies in hepatitis C virus infected cirrhotic patients from southern Italy. AB - Non-organ-specific autoantibodies (NOSAs) are frequently found in patients with hepatitis C virus (HCV) chronic infection. Genetics is likely involved in the development of autoimmune reactivities, and differences in the prevalence of HCV related autoantibodies among populations of various geographic areas should be expected. We evaluated the prevalence and the clinical impact of NOSAs in a series of HCV-infected patients from southern Italy. We studied 283 consecutive anti-HCV positive patients (162 men, 121 women, mean age 54.5 +/- 13.5 years), 94 of whom were cirrhotics and 189 noncirrhotics. Serum from each patient and from 41 hepatitis B surface antigen (HBsAg)-positive/anti-HCV negative control subjects were tested (dilution 1:40) for autoantibodies by indirect immunofluorescence. Qualitative/quantitative HCV-RNA determinations were also performed. The prevalence of NOSAs was significantly higher in anti-HCV-positive subjects than in HBsAg-positive patients (P < 0.006). Autoantibodies were significantly associated with both cirrhosis (P < 0.0001) and older age (P < 0.05). No significant association between NOSAs and either female gender or virological parameters (HCV-RNA positivity, viral load, and genotype) was found. In conclusion, the autoantibody positivity in HCV-infected patients from southern Italy is significantly related to cirrhosis and older age, although its general prevalence is similar to that reported in populations from the north of the country. PMID- 12643615 TI - Propafenone hepatotoxicity: report of two new cases. AB - Two patients developed acute cholestatic hepatitis during treatment with propafenone. Viral infections, alcohol abuse, hepatotoxicity by other drugs, and biliary obstruction were excluded as causes. In one patient, liver biopsy showed changes consistent with a drug-associated injury. Another patient had autoimmune antibodies (ANA) in the serum. Following propafenone withdrawal, the clinical and biochemical profiles of both patients improved. Hepatic toxicity from the antiarrhythmic drug propafenone is highly uncommon. Moreover, the drug produces hepatocellular injury by an unknown mechanism. Most of the seven cases reported here had acute cholestatic hepatitis after a latency period of two to four weeks. PMID- 12643613 TI - Overexpression of heat shock protein 70 in stomach of stress-induced gastric ulcer-resistant rats. AB - Expression of heat shock protein 70, induced by an antiulcer drug, provides protection against gastric ulcers. However, the mechanisms responsible for this protection are not known. The expression in ulcer-resistant, spontaneously hypertensive rats was 2.8-fold higher than in normotensive rats. One hour after restraint and water immersion stress, strong nuclear immunoreactivity was observed in nuclei of surface epithelial cells at the crest of gastric mucosal folds, the first site of ulceration, only in spontaneously hypertensive rats. Heat shock cognate protein 70, which is expressed in mucus-secreting cells, was not overexpressed in spontaneously hypertensive rats. Heat shock protein 70 expression was attenuated by chemical sympathectomy, which also resulted in abolition of the increase of mucosal blood flow and aggravation of ulcers. Our results indicate that overexpression of heat shock protein 70 in the stomach seems to protect against gastric ulcers through its cytoprotective effects on gastric mucosa by increasing mucosal blood flow. PMID- 12643616 TI - Hepatic hydatid cyst causing thrombosis of the inferior vena cava and complicated by hemobilia: a multimodal sequential approach in the treatment. PMID- 12643617 TI - Reciprocal variations of nNOS and HSP90 are associated with fasting in gastrointestinal tract of the piglet. AB - The effects of fasting on neuronal NO synthase (nNOS), Heme oxygenase 2 (HO-2), and heat shock protein 90 (HSP90) was determined by immunoblotting in the stomach, duodenum, mid-jejunum, distal ileum, and proximal colon of 28-day-old piglets. nNOS expression was drastically reduced in all the gastrointestinal areas studied while HO-2 was not changed. Concomitant with the nNOS decrease, elevated expressions of HSP90 were observed in these different areas. These results are discussed in terms of the regulation relationship between NOS and HSP90 and the possible protective effect of the heat shock protein and the potential application in digestive pathologies. PMID- 12643618 TI - Replacement of conventional glucocorticoids by oral pH-modified release budesonide in active and inactive Crohn's disease: results of an open, prospective, multicenter trial. AB - Glucocorticosteroids (GCS) are established in the treatment of active Crohn's ileitis and ileocolitis. Recently, the topical steroid budesonide was found to be effective in untreated patients with Crohn's disease (CD) causing less side effects than conventional GCS. No clinical data have been reported about the effects of switching from conventional GCS to budesonide in terms of side effects and disease activity. The primary aim of this study was to evaluate the development of side effects after switching from conventional GCS treatment to Eudragit L-coated budesonide (pH-modified release formulation) in patients taking 5-30 mg prednisolone equivalent per day for at least two weeks. In all, 178 patients with active CD (N = 88) or CD in remission during GCS treatment (N = 90) were included. Conventional GCS treatment was tapered down during a maximum of three weeks, with simultaneous intake of 3 x 3 mg budesonide. Thereafter, patients received 3 x 3 mg budesonide alone for six weeks. GCS-related side effects, disease activity and adverse events were documented at study entry and after 0, 2, 4, and 6 weeks of budesonide treatment. The percentage of patients with GCS-related side effects decreased from 65.2% (intention-to-treat population) at entry to 43.3% (P < 0.0001) at the end of the trial. The total number of GCS-related side effects decreased significantly from 269 to 90. Of the patients who entered the study with active disease under conventional GCS therapy, 38.6% were in remission at the end of the study. Of the patients who entered the study with CD in remission, 78% stayed in remission after switching from conventinal GCS to budesonide. In conclusion, switching from conventional GCS treatment to budesonide leads to a significant reduction of GCS related side effects in patients with CD without causing rapid deterioration of the disease. PMID- 12643619 TI - Growth promoting effect of thioredoxin on intestinal epithelial cells. AB - Paneth cells are located at the bases of intestinal crypts, and their cytoplasmic granules contain large amounts of zinc. We previously showed that administration of diphenylthiocarbazone (dithizone), a zinc chelater, to rats induced the selective death of Paneth cells. This was followed by a transient wave of epithelial cell proliferation in the entire crypts. In the study described here, we again applied this experimental model in an attempt to identify novel growth promoting factors in the small intestine. Male Wistar rats were injected with dithizone and killed 6 hr later. Messenger RNAs (mRNAs) were extracted from the terminal ileum for the construction of a cDNA library. This library was then transfected into the human intestinal cell line Caco-2, and the cells that continued to grow in the medium containing only 1% FBS were cloned. One of the cDNA sequences identified from those transfection experiments was the full-length rat thioredoxin (TRX) gene. To confirm the growth-promoting effect of this cDNA, we transfected it into Caco-2 cells again. These clones proliferated in the medium containing only 1% FBS, while the control clones failed to show any growth. Transient oxidative stress exerted by the addition of oxidative reagents diamide and hydrogen peroxide partially suppressed the growth of TRX-transfected cells. Northern hybridization analysis revealed that TRX expression in rat ileum after dithizone treatment was altered in accordance with intestinal epithelial regeneration in the crypts. Single-cell RT-PCR also showed TRX mRNA expression in Paneth cells. These studies identify rat thioredoxin as a growth-promoting factor for intestinal epithelial cells. PMID- 12643621 TI - Changing presentation of adult celiac disease. AB - The mode of presentation of celiac disease in the United States is not known. We investigated the demographic and clinical features of 227 patients with biopsy proven celiac disease and determined if there had been changes over time. The patients had been entered into a database; those seen prior to 1990 were retrospectively entered while those seen subsequently were prospectively entered. A "symptomatic" presentation described the "classical" presentation of celiac disease with prominent gastrointestinal symptoms: diarrhea and weight loss. Females were younger and had a longer duration of symptoms compared to males. The modes of presentation were symptomatic (62%), anemia or reduced bone density (15%), screening first-degree relatives (13%), and incidental diagnosis at endoscopy (8%). We compared those diagnosed before and after 1993 (when serologic testing was first used), and noted a reduction in those presenting with diarrhea, 73% vs 43% (P = 0.0001) and a reduction in the duration of symptoms, from 9.0 +/- 1.1 years to 4.4 +/- 0.6 years (P < 0001). In conclusion, the percentage of celiac disease patients presenting with diarrhea has decreased, probably related to the more widespread use of serologic testing for celiac disease. PMID- 12643620 TI - Anxiety and depression are related to autonomic nervous system function in women with irritable bowel syndrome. AB - This study compared women with irritable bowel syndrome who had a history of an anxiety or depressive disorder to those without symptoms of either disorder on indicators of cardiac parasympathetic activity, autonomic nervous system balance, and general autonomic activity. The Diagnostic Interview Schedule was used to determine anxiety or depressive disorders, and a Holter monitor was used to record R-R intervals over 24 hr. A similar comparison was done with healthy controls. Among women with irritable bowel syndrome, those with a positive history had lower parasympathetic and general activity throughout the 24-hr period than did women without a diagnosis. Indicators of autonomic balance were slightly higher in women with a positive history compared to those without a history. Similar differences were seen in controls. Thus, a history of anxiety and depressive disorders is associated with lower parasympathetic activity, both in women with IBS and healthy controls. Further exploration is needed to understand if lower parasympathetic activity influences the pain and stool pattern changes seen in persons with irritable bowel syndrome. PMID- 12643622 TI - Prevention of acetic acid-induced colitis by desferrithiocin analogs in a rat model. AB - Iron contributes significantly to the formation of reactive oxygen species via the Fenton reaction. Therefore, we assessed whether a series of desferrithiocin analogs, both carboxylic acids and hydroxamates, could (1) either promote or diminish the iron-mediated oxidation of ascorbate, (2) quench a model radical species, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+), and (3) when applied topically, prevent acetic acid-induced colitis in rats. Surprisingly, most of the desferrithiocin analogs inhibited the Fenton reaction to an approximately equivalent degree; however, substantial differences were observed in the capacity of the analogs to scavenge the model radical cation. Four carboxylic acid desferrithiocin analogs and their respective N methylhydroxamates were tested along with desferrioxamine and Rowasa, a currently accepted topical therapeutic agent for inflammatory bowel disease (IBD), in a rodent model of acetic acid-induced colitis. The colonic damage was quantitated by two independent measurements. Although neither radical scavenging nor prevention of Fenton chemistry was a definitive predictor of in vivo efficacy, the overall trend is that desferrithiocin analogs substituted with an N methylhydroxamate in the place of the carboxylic acid are both better free radical scavengers and more active against acetic acid-induced colitis. These results represent an intriguing alternative avenue to the development of improved IBD therapeutic agents. PMID- 12643623 TI - Elimination of local macrophages in intestine prevents chronic colitis in interleukin-10-deficient mice. AB - Uncontrolled activation of T cells and macrophages is involved in the development of inflammatory bowel disease (IBD). However, the precise role of intestinal macrophages for development of IBD is till unclear. To investigate the role of local macrophages for development of IBD, we developed poly-D, L-lactic acid microspheres containing dichloromethylene diphosphonate, which was specifically taken up by macrophages and depleted them, and then the animal model for human IBD was treated with this reagent. We have shown that rectal administration of the microspheres reduced the numbers of resident macrophages in the intestinal lymphoid follicles of interleukin-10-deficient mice. Importantly, depletion of intestinal macrophages was associated with suppression of development of chronic colitis. These results suggest that local macrophages in the intestine play a critical role in the development of chronic colitis in an animal model for IBD. Our study implies that elimination of resident macrophages in the intestine may become a therapeutic approach to IBD. PMID- 12643624 TI - Pancreatitis and E. coli O157:H7 colitis without hemolytic uremic syndrome. PMID- 12643625 TI - Liver abscess and diarrhea as initial manifestations of ulcerative colitis: case report and review of the literature. PMID- 12643626 TI - Determination of the hydroperoxil group level in polyethylene glycols using a novel mimic enzyme Fe(III)-Mn(II)(HNAPTS)2. AB - Two kinds of Schiff base metal complexes of 2-hydroxy-1-naphthaldehyde-2-amino-5 phenylthiazole (HANPTS), Mn(II)(HNAPTS)(2) and Fe(III)(HNAPTS)(2), were synthesized and used to mimic the active group of horseradish peroxidase (HRP). The catalytic characteristics of the mimic enzymes in the oxidation reaction of ascorbic acid (AsA) with the OOH group in polyethylene glycols (PEGs) have been studied by a spectrophotometric method. Fe(III) has remarkable coordinated catalysis to Mn(II)(HNAPTS)(2); as a result, the catalytic ability of Fe(III) Mn(II)(HNAPTS)(2) is 75% of that of HRP. The possible mechanism of the reaction was discussed. The linear relationship between deltaA(265)(AsA) and OOH group concentrations was in the range of 1.5 x 10(-6) to 9.0 x 10(-4) mol/L. The proposed method was successfully applied to the determination of the OOH group level in different molecular weight PEGs. PMID- 12643628 TI - Establishment and application of a fluorescent polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method for identifying porcine, caprine, and bovine meats. AB - A method of fluorescent Polymerase Chain Reaction-restriction fragment length polymorphism (PCR-RFLP) was applied as an analytical and quantitative tool for meat identification. Following alignments of the nucleotide sequences, an oligonucleotide primer pair was designed to amplify the partial sequences within the 12S ribosomal RNA (12S rRNA) gene of mitochondrial DNA from porcine, caprine, and bovine meats. No fragment can be amplified from dog, cat, fish, duck, goose, turkey, and chicken DNA with the primer pair. Using fluorescence sensor capillary electrophoresis, the species-specific DNA fingerprints of pork, goat, and beef were generated by restriction enzyme digestion following a fluorescence-labeling PCR amplification. Species identification was conducted on the meat mixtures. The reliably semiquantitative levels were below 1% for binary mixtures of pork, goat, and beef. Cooking and autoclaving of meats did not influence the generation of the PCR-RFLP profiles or the analytical accuracy. PMID- 12643627 TI - Identification of processed Japanese green tea based on polymorphisms generated by STS-RFLP analysis. AB - In Japan, tea is generally sold blended, though 90% of the total production is clonal. Due to the increasingly strict consumer need and taste, however, more and more Japanese green teas are being sold under their particular cultivar name. Moreover, tea made from Yabukita, a much appreciated cultivar originally developed in Japan, has recently been produced and imported from a neighboring country. This paper describes a simple and inexpensive methodology capable of identifying fresh and processed Japanese green teas to discourage its fraudulent commercialization. The study was based on 46 main tea cultivars, and polymorphism detected through STS-RFLP analysis of the coding and noncoding DNA regions of three genes, namely phenylalanine ammonia-lyase, chalcone synthase, and dihydroflavonol 4-reductase, for which nucleotide information was available. All 46 tea cultivars analyzed could be easily distinguished using a combination of codominant DNA markers. Yabukita displayed a unique profile when PAL intron was digested with DdeI, thus allowing its rapid authentication at low cost. PMID- 12643629 TI - Elucidation of pectin methylester distributions by capillary electrophoresis. AB - Evidence is presented that the electrophoretic mobility of the polysaccharide pectin in typical capillary electrophoresis (CE) experiments is determined largely by its chain-averaged charge density, irrespective of how that charge is distributed. This was found to be the case for both high and low calcium sensitive sister fractions separated from a mother sample on the grounds of calcium affinity and for blocky methylester distributions generated by a processive demethylating enzyme. The blockiness of the generated intramolecular methylester distribution was substantiated experimentally, also by CE, by observing the oligomeric digest fragments produced by incubation with endo polygalacturonase II from Aspergillus niger. The conclusion that the CE migration time is largely invariant to the intramolecular methylester distribution supports the idea that the CE peak shapes can be used to give a useful indication of the intermolecular methylester distribution. PMID- 12643630 TI - Gas chromatography-olfactometry (GC-O) and proton transfer reaction-mass spectrometry (PTR-MS) analysis of the flavor profile of grana padano, parmigiano reggiano, and grana trentino cheeses. AB - Gas chromatography-olfactometry (GC-O) and proton transfer reaction-mass spectrometry (PTR-MS) techniques were used to deduce the profile of odor-active and volatile compounds of three grana cheeses: Grana Padano (GP), Parmigiano Reggiano (PR), and Grana Trentino (GT). Samples for GC-O analysis were prepared by dynamic headspace extraction, while a direct analysis of the headspace formed over cheese was performed by PTR-MS. The major contributors to the odor profile were ethyl butanoate, 2-heptanone, and ethyl hexanoate, with fruity notes. A high concentration of mass 45, tentatively identified as acetaldehyde, was found by PTR-MS analysis. Low odor threshold compounds, e.g., methional and 1-octen-3-one, which contributed to the odor profile but were not detected by FID, were detected by PTR-MS. Principal component analysis on both GC-O and PTR-MS data separated the three cheese samples well and showed specific compounds related to each sample. PMID- 12643631 TI - Rapid determination of domoic acid in serum and urine by liquid chromatography electrospray tandem mass spectrometry. AB - A rapid, selective, and sensitive LC-MS/MS method was developed for the quantitative determination of domoic acid in serum and urine samples. Samples were prepared for analysis using an Oasis HLB SPE column. Determination was by a reversed phase HPLC using a mixture of methanol, acetonitrile, and water containing 1% acetic acid and an electrospray ionization (ESI) ion-trap mass spectrometer (Finnigan LCQ). The method was validated by analyzing five replicates each of negative control bovine serum or urine fortified with domoic acid at the 0.005 microg/g method detection limit (MDL) and at the 0.05 microg/g level. Recoveries ranged from 90 to 95% for fortifications at the MDL and from 92 to 98% for fortifications 10 times higher than the MDL. The diagnostic utility of the method was tested by analyzing samples from live animals showing clinical signs suggestive of domoic acid poisoning submitted to the veterinary toxicology laboratory. PMID- 12643632 TI - Stability of saponins in alcoholic solutions: ester formation as artifacts. AB - Saponins containing a free carboxylic group in the molecule give the corresponding esters as artifacts when stored for a long time in alcoholic solutions. Two saponins from Medicago sativa L., chosen on the basis of their different positions of the carboxylic group in the molecule, were refluxed with methanol and ethanol under neutral conditions. 3,28-di-O-glu medicagenic acid possesses a carboxylic group on the triterpenic moiety, whereas soyasaponin I, a glycoside of soyasapogenol B, has a glucuronic acid unit as the first sugar linked to the triterpene structure. Artifacts were quantified by HPLC. The peaks identified as the corresponding esters were examined during boiling from 1 h to 5 days. Quantitative results indicated that the carboxylic group on the sugar moiety, as for soyasaponin I, is more reactive than that on the triterpenic structure, as for 3,28-di-O-glu medicagenic acid. Saponins having the free carboxylic groups create enough acidity in their alcoholic solutions to catalyze the formation of the corresponding esters. PMID- 12643633 TI - Detection of sugar syrups in apple juice by delta(2)H per thousand and delta(13)C per thousand analysis of hexamethylenetetramine prepared from fructose. AB - An improved procedure for determining (13)C and (2)H isotope ratios, using gas chromatography-isotope ratio mass spectrometry (GC-IRMS), has been developed for identifying the addition of low cost commercial sugar syrups to apple juices and related products. Isotopic techniques are commonly used to identify the addition of low cost sugars to fruit juices and are difficult to circumvent as it is not economically viable to change the isotopic ratios of the sugars. The procedure utilizes the derivative hexamethylenetetramine, which is produced through chemical transformation of a sugar degradation product and provides position specific (13)C and (2)H ratios that relate to the parent sugar molecule. The new procedure has advantages over methods using nitro-sugar derivatives in terms of analysis time and sensitivity. The differences between the delta(2)H per thousand and delta(13)C per thousand values of the 100 authentic apple juices and beet and cane commercial sugar syrups permit their addition to be reliably detected. PMID- 12643634 TI - Optimization of microwave-assisted extraction (MAP) for ginseng components by response surface methodology. AB - Response surface methodology (RSM) was applied to predict optimum conditions for microwave-assisted extraction-a MAP technology-of saponin components from ginseng roots. A central composite design was used to monitor the effect of ethanol concentration (30-90%, X(1)) and extraction time (30-270 s, X(2)) on dependent variables, such as total extract yield (Y(1)), crude saponin content (Y(2)), and saponin ratio (Y(3)), under atmospheric pressure conditions when focused microwaves were applied at an emission frequency of 2450 MHz. In MAP under pre established conditions, correlation coefficients (R (2)) of the models for total extract yield and crude saponin were 0.9841 (p < 0.001) and 0.9704 (p < 0.01). Optimum extraction conditions were predicted for each variable as 52.6% ethanol and 224.7 s in extract yield and as 77.3% ethanol and 295.1 s in crude saponins, respectively. Estimated maximum values at predicted optimum conditions were in good agreement with experimental values. PMID- 12643636 TI - Polychlorinated biphenyls (PCBs) detection in food samples using an electrochemical immunosensor. AB - In this work, a disposable electrochemical immunosensor, based on a competitive assay scheme, was applied to detect polychlorinated biphenyls (PCBs) in food. For this purpose, antibodies against PCBs were directly immobilized onto the carbon surface of a disposable screen-printed electrode. A competition between the PCBs present in the sample and a fixed concentration of an enzyme-labeled PCB was realized and evaluated by electrochemical detection. Alkaline phosphatase was used as the enzyme label, coupled with differential pulse voltammetry (DPV) as the electrochemical technique. The immunosensor was tested on aroclor mixture detection (1242 and 1248) and then on some typologies of food samples to evaluate the possible application for real sample analysis. Samples analyzed were from different matrixes, such as sheep milk, bovine adipose tissue, and bovine muscle. Results obtained were compared with the accredited results according to ISO 17025 methods for PCB detection (HRGC-LRMS) as a confirmatory analysis. Preliminary results show the possibility to use this device as a screening method in food sample analysis. The negligible matrix effect observed may lead to a simplified extraction procedure, and considerable time and consumable savings are the immediate benefits given by the proposed method. PMID- 12643635 TI - Utilization of Folin-Ciocalteu phenol reagent for the detection of certain nitrogen compounds. AB - To determine in more detail the reaction of Folin-Ciocalteu phenol reagent with nitrogen compounds, a number of hydroxylamine-related compounds and a large number of guanidine-containing compounds were tested. In general, guanidine compounds did not react strongly unless they were hydroxyamino or hydrazino derivatives. The non-hydroxyamino paralytic shellfish poison saxitoxin, however, reacted to a significant extent. This may be due to the presence of a five membered ring structure and its analogy to 2-aminopurines, which react strongly. A number of simpler amines were also tested. Tertiary aliphatic amines, but not primary, secondary, or quaternary amines, reacted strongly with the reagent. Primary, secondary, and tertiary aromatic amines all reacted strongly with the reagent. Reactivity was extended to pyrroles and indole derivatives but not to imidazole and benzimidazole derivatives. Defining the reactivity of Folin Ciocalteau phenol reagent with nitrogen compounds extends the usefulness of the reagent for the detection and determination of certain nitrogen compounds in basic extracts by colorimetric means and by thin-layer chromatography. PMID- 12643637 TI - Development of an enzyme-linked immunosorbent assay for the insecticide thiamethoxam. AB - An enzyme-linked immunosorbent assay (ELISA) was developed for the neonicotinoid insecticide thiamethoxam, 3-(2-chlorothiazol-5-ylmethyl)-5-methyl-4-nitroimino 1,3,5-oxadiazinane. Three antisera were raised from rabbits immunized with the hapten-KLH conjugate. On the basis of the computational analysis of hapten candidates, the hapten with a spacer arm on the thiazolyl ring of thiamethoxam was synthesized to elicit thiamethoxam-specific antisera. The hapten was 3-[2-(2 carboxyethylthio)-5-ylmethyl]-5-methyl-4-nitroimino-1,3,5-oxadiazinane. Antisera were characterized with indirect competitive ELISA. Cross-reactivity and effects of organic solvents, pH, and ionic strengths were evaluated. The antiserum was specific for thiamethoxam and tolerant of up to 5% acetonitrile and 5% acetone. Various ionic strengths and pH values in the tested ranges had negligible effect on the assay performance. Under the optimized conditions, the half-maximal inhibition concentration (IC(50)) and the limit of detection were approximately 9.0 and 0.1 microg/L of thiamethoxam, respectively. ELISA analysis of stream and tap water samples showed an excellent correlation with the fortification levels. PMID- 12643638 TI - Simple detection method of powerful antiradical compounds in the raw extract of plants and its application for the identification of antiradical plant constituents. AB - A simple detection method for a powerful radical scavenging compound in a mixture containing a large variety of compounds, such as the raw extract of edible plants, was developed using 1,1-diphenyl-2-picrylhydrazyl (DPPH) as the radical reagent. The method was established on the basis of the features of the typical chain-breaking antioxidation reaction mechanism, which suggests that the radical scavenging antioxidant should be converted to other stable nonradical compounds during the reaction. This method requires only a simple HPLC instrument, and the disappearance or decrease in the peak intensity, which is induced by the addition of DPPH. This change is monitored by the HPLC to detect the powerful radical scavenger from the complex mixture. The method was applied to the detection and identification of the most powerful antiradical compound in the extracts of three antioxidatively active plant extracts (Psidium guajava, Citrus depressa, and Hypericum chinense). The radical scavenging efficiency of a newly identified compound from H. chinense was also compared with that of Trolox and catechin using the method. PMID- 12643639 TI - Characterization of anthocyanins in grape juices by ion trap liquid chromatography-mass spectrometry. AB - A reverse phase HPLC and electrospray interface with ion trap mass spectrometer method was developed for the characterization of anthocyanins in Concord, Rubired, and Salvador grape juices. Rubired and Salvador grapes are hybrids from Vitis vinifera and Vitis rupestris. Concord grape is a grape from the native American cultivar Vitis labrusca. Individual anthocyanins in these three varieties were identified on the basis of UV-vis and MS spectra and further elucidated by MS/MS spectra. Anthocyanins in Salvador and Concord grapes were 3-O glucosides, 3-O-(6' '-O-p-coumaroyl)glucosides, 3-O-(6' '-O-p-acetyl)glucosides, 3,5-O-diglucosides, and 3-O-(6' '-O-p-coumaroyl)-5-O-diglucosides of delphinidin, cyanidin, petunidin, peonidin, and malvidin. Vitisin B was detected in Salvador grape juice. Anthocyanins in Rubired grape juice were primarily anthocyanin diglucosides: peonidin 3,5-O-diglucoside, malvidin 3,5-O-diglucoside, peonidin 3 O-(6' '-O-p-coumaroyl)-5-O-diglucoside, and malvidin 3-O-(6' '-O-p-coumaroyl)-5-O diglucoside are the four major anthocyanins. The presence of pelargonidin 3-O glucoside, not previously reported, has been established for the first time in all three juices. PMID- 12643640 TI - Solanidine hydrolytic extraction and separation from the potato (Solanum tuberosum L.) vines by using solid-liquid-liquid systems. AB - Solanidine is a steroidal aglycon of potato (Solanum tuberosum L.) glycoalkaloids and a very important precursor for the synthesis of hormones and some pharmacologically active compounds. Glycoalkaloids are hydrolyzed by mineral acid, yielding solanidine. This paper deals with the kinetics of solanidine hydrolytic extraction in different solid-liquid-liquid systems. The dried and milled potato (S. tuberosum L.) vines were used as a source of glycoalkaloids and as the solid phase. The solutions of hydrochloric acid in 2 and 10% (w/v) aqueous acetic acid, in 50% (volume) aqueous methanol, and in 50% (volume) aqueous ethanol were first liquid phase, and the medium for glycoalkaloid extraction from potato vines and their hydrolysis to solanidine. The chloroform, trichloroethylene, or carbon tetrachloride were the second, organic, liquid phase and the medium for solanidine extraction. This procedure combines three different processes: extraction of glycoalkaloids from potato vines, their hydrolysis to solanidine, and the extraction of solanidine, in a single step. The term hydrolytic extraction of solanidine was used for these processes. The purpose of the paper was to choose an optimal solid-liquid-liquid system for solanidine extraction and to define the procedure for its isolation from the organic liquid phase. The best degree of solanidine hydrolytic extraction (DHE) of more than 98% was achieved when 10% (w/v) hydrochloric acid in 50% (volume) methanol were the first liquid phase and chloroform was the second liquid phase, after 90 min. The yield of solanidine (q(S)) under these conditions is calculated to be 0.24 g/100 g of potato vines. Approximately 78% of the maximal possible yield of solanidine was isolated from chlorofom liquid phase. The IR and MS spectra of isolated solanidine were recorded. PMID- 12643641 TI - Estrogenic activity in white and red wine extracts. AB - Red wine is enriched in resveratrol, trans-3,5,4'-trihydroxystilbene, a compound in grape skin that inhibits the development of pre-neoplastic lesions in mouse mammary tumor cells in culture and inhibits cancer cell proliferation in vitro. Grapes also contain other bioactive compounds including flavonoids, flavans, and anthocyanins. The estrogenic activities of extracts prepared from one white (Freie Weingartner Wachau, Gruner Veltliner, Austria) and two red wines (Woodbridge, Cabernet Sauvignon, California; and Lenz Moser Prestige, Blaufrankisch Barrique, Austria) were examined and compared with those induced by estradiol (E(2)) and trans-resveratrol. First, the estrogenic activity of the wine extracts was evaluated in a yeast estrogen screen (YES) assay, in which yeast express copper-inducible estrogen receptor alpha (ERalpha) and an estrogen response-element (ERE)-driven beta-galactosidase reporter. In YES, the white wine extract showed no estrogenic activity. In contrast, both of the red wine extracts showed estrogenic activity equivalent to that of 0.2 nM E(2). Similarly, the white wine extract showed no transcriptional activity with either ERalpha and ERbeta in transiently transfected CHO-K1 cells. In contrast, both red wine extracts stimulated ERE-reporter activity in a concentration-dependent manner that was inhibited by 4-hydroxytamoxifen (4-OHT), indicating that the observed transcriptional activity was ER-mediated. The red wine extracts showed significantly higher ERbeta versus ERalpha agonist activity. Resveratrol showed no agonist activity in YES but activated ERalpha and ERbeta in CHO-K1 cells in a concentration-dependent manner that was inhibited by 4-OHT. This indicates that resveratrol requires mammalian cell components that are absent in yeast for estrogen agonist activity, whereas the estrogenic activity of wine extracts is directly through ERalpha and does not require mammalian cell factors such as coactivators. The estrogenic activity in red wine found by using YES indicates that estrogenic compounds other than resveratrol are present. Chemical analysis clearly showed that the trans-resveratrol content of the red wine extracts was 1 order of magnitude below the detection limit for YES assay. PMID- 12643643 TI - Factors affecting the levels of tea polyphenols and caffeine in tea leaves. AB - An isocratic HPLC procedure was developed for the simultaneous determination of caffeine and six catechins in tea samples. When 31 commercial teas extracted by boiling water or 75% ethanol were analyzed by HPLC, the levels of (-) epigallocatechin 3-gallate (EGCG), and total catechins in teas were in the order green tea (old leaves) > green tea (young leaves) and oolong tea > black tea and pu-erh tea. Tea samples extracted by 75% ethanol could yield higher levels of EGCG and total catechins. The contents of caffeine and catechins also have been measured in fresh tea leaves from the Tea Experiment Station in Wen-Shan or Taitung; the old tea leaves contain less caffeine but more EGCG and total catechins than young ones. To compare caffeine and catechins in the same tea but manufactured by different fermentation processes, the level of caffeine in different manufactured teas was in the order black tea > oolong tea > green tea > fresh tea leaf, but the levels of EGCG and total catechins were in the order green tea > oolong tea > fresh tea leaf > black tea. In addition, six commercial tea extracts were used to test the biological functions including hydroxyl radical scavenging, nitric oxide suppressing, and apoptotic effects. The pu-erh tea extracts protected the plasmid DNA from damage by the Fenton reaction as well as the control at a concentration of 100 microg/mL. The nitric oxide suppressing effect of tea extracts was in the order pu-erh tea >/= black tea > green tea > oolong tea. The induction of apoptosis by tea extract has been demonstrated by DNA fragmentation ladder and flow cytometry. It appeared that the ability of tea extracts to induce HL-60 cells apoptosis was in the order green tea > oolong > black tea > pu-erh tea. All tea extracts extracted by 75% ethanol have stronger biological functions than those extracted by boiling water. PMID- 12643642 TI - Association of suppression of extracellular signal-regulated kinase phosphorylation by epigallocatechin gallate with the reduction of matrix metalloproteinase activities in human fibrosarcoma HT1080 cells. AB - Matrix metalloproteinases (MMPs) play a crucial role in the process of cancer invasion and metastasis. Previous findings suggested that epigallocatechin gallate (EGCG), a main flavanol of green tea, caused decreased levels of MMP-2 and MMP-9 activities to be secreted into culture medium. To obtain further information on EGCG-mediated regulation of these MMPs, the effects of EGCG on enzyme activity, mRNA expression, and mitogen-activated protein kinase (MAPK) activities in human fibrosarcoma HT1080 cells were examined. EGCG was confirmed to suppress the gelatin-degrading activities due to MMP-2 and MMP-9 in the culture medium. This suppression of enzyme activities by EGCG was consistent with the decreased levels of MMP-2 and MMP-9 mRNAs. EGCG-mediated suppression was also observed for MT1-MMP mRNA. EGCG-mediated suppression of the level of MMP-9 transcript was correlated with its suppression of MMP-9 promoter activity. EGCG inhibited the phosphorylation of extracellular signal-regulated kinases 1 and 2 (ERK1/2), which are the members of an MAPK family necessary for MMP-9 up regulation. EGCG also suppressed p38 MAPK activity but gave no effects on stress activated protein kinase/c-Jun N-terminal kinase activity. These findings suggest that suppression of ERK phosphorylation by EGCG is involved in the inhibition of expression for MMP-2 and MMP-9 mRNAs, leading to the reduction of their enzyme activities of the cancer cells. Methyl derivatives, epigallocatechin-3-O-(3-O methyl) gallate and epigallocatechin-3-O-(4-O-methyl) gallate, exhibited effects similar to, but weaker than, those of EGCG, suggesting the important role of an unsubstituted triphenolic ester structure in these activities. PMID- 12643644 TI - Structure-antioxidant activity relationship of ferulic acid derivatives: effect of carbon side chain characteristic groups. AB - Differences in the antioxidant activity of some biosynthetically related ferulic acid derivatives induced by the presence of characteristic groups (-COOH, -CHO, CH(2)OH, -CH(3), and -COOC(2)H(5)) at the end of their carbon side chain were investigated using both experimental and computational methods. The relative order of the scavenging activity toward the DPPH radical derived from kinetic studies was isoeugenol approximately coniferyl alcohol >> ferulic acid approximately coniferyl aldehyde approximately ethyl ferulate. In bulk oil autoxidation (45 degrees C) the same order of activity was obtained. In the o/w emulsion autoxidation, lipophilicity of the phenols was the determining factor because the least polar compounds bearing -CH(3) and -COOC(2)H(5) were the most effective ones. The order of activity based on the O-H bond dissociation enthalpy (BDE) and ionization potential (IP) values, calculated by the density functional theory (DFT) method, was in accordance with the experimental radical scavenging order and with the electron-donating/withdrawing properties of the characteristic groups. Other molecular descriptors could not complement the experimental findings. PMID- 12643645 TI - Lytic activity of l-menthol derivatives against the snow blight disease fungus, Micronectriella nivalis. AB - Lytic activity of l-menthol (1) derivatives [(-)-(1S,3R,4S,6S)-6-hydroxymenthol (2), (-)-(1S,3R,4S)-1-hydroxymenthol (3), and (+)-(1S,3R,4R,6S)-6,8 dihydroxymenthol (4)] against the snow blight disease fungus, Micronectriella nivalis was investigated. Compounds 2, 3, and 4 had 85.0, 63.9, and 81.9% lytic activity, respectively, at a concentration of 0.2 mg/mL. The activity of each of the three compounds increased in a dose-response manner. To study the structure activity relationship, acetyl esters of 1-4 [(-)-menthyl acetate (1Ac), (-)-6 hydroxymenthyl diacetate (2Ac), (-)-1-hydroxymenthyl 3-monoacetate (3Ac), and (-) 6,8-dihydroxymenthyl 3,6-diacetate (4Ac)] were synthesized with yields of 80.2 99.8% and were also assayed. The acetyl esters of 1Ac, 2Ac, 3Ac, and 4Ac had 51.2, 91.5, 66.0, and 95.2% lytic activity, respectively, at a concentration of 0.2 mg/mL, and these compounds showed further high lytic activity compared with the alcohols of 1-4. These acetyl esters also showed higher lytic activity as their concentration was increased. Of particular interest is the fact that 2Ac and 4Ac both had higher lytic activity at 0.05-0.2 mg/mL compared with copper 8 hydroxyquinolate, a standard chemical widely used to control snow blight. This is the first report on lytic activity of l-menthol derivatives. PMID- 12643646 TI - Water activity effects on geranyl acetate synthesis catalyzed by novozym in supercritical ethane and in supercritical carbon dioxide. AB - The esterification reaction of geraniol with acetic acid catalyzed by Novozym was studied in supercritical ethane (sc-ethane) and in supercritical carbon dioxide (sc-CO(2)). Water activity (a(W)) had a very strong effect on enzyme activity, with reaction rates increasing up to a(W) = 0.25 and then decreasing for higher a(W). Salt hydrate pairs could not prevent changes in a(W) during the course of reaction but were able to control a(W) to some extent and had a beneficial effect on both initial rates of esterification and conversion in sc-ethane. The enzyme was more active in sc-ethane than in sc-CO(2), confirming the deleterious effect of the latter already observed with some enzymes. Temperatures between 40 and 60 degrees C did not have a strong effect on initial rates of esterification, although reaction progress declined considerably in that temperature range. For the mixture of 50 mM acetic acid plus 200 mM geraniol, 100% conversion was achieved at a reaction time of 10 h at 40 degrees C, 100 bar, an a(W) of incubation of 0.25, and a Novozym concentration of 0.55 mg cm(-)(3) in sc-ethane. Conversion was below 50% in sc-CO(2) at otherwise identical conditions. With an equimolar mixture of the two substrates (100 mM), 98% conversion was reached at 10 h of reaction in sc-ethane (73% conversion in sc-CO(2)). PMID- 12643647 TI - Optimization of making barrel-fermented dry muscatel wines. AB - The optimization of making barrel-fermented muscatel wines requires determining what type of must clarification is most suitable for the quality of the wine, as well as what type of barrel will yield the most acceptable wines. This is achieved by adding pectolytic enzymes to clarify part of the muscatel must statically; the rest is clarified by vacuum filtration. The musts obtained are fermented in French and American oak barrels and, once fermentation has ceased, they are kept with their lees for 2 months, with periodic stirring. Eleven conventional parameters and 31 volatile compounds were quantified, and a sensory analysis of the wines was produced, which led us to conclude that static clarification with pectolytic enzymes from the muscatel musts produces the best structured wines and the larger content of higher alcohols, esters, and terpenic compounds. The wines fermented in American oak barrels received the highest overall marks, which may be due to the greater aromatic complexity given off by the compounds in the wood. PMID- 12643648 TI - Effect of high pressure on isomerization and degradation of lactose in alkaline media. AB - The combined effects of temperature (60 degrees C) and high-pressure treatments (400 MPa for 3 h) on the isomerization-degradation of lactose (10%) in basic media were studied. The formation of isomeric disaccharides (lactulose and epilactose) and galactose decreased by the application of high pressure in aqueous sodium hydroxide 4 mM (pH 10.2) and 8 mM (pH 10.6), and sodium carbonate bicarbonate buffer (pH 10.0). In addition, no substantial color development was observed in the sodium hydroxide systems for high and atmospheric pressure, whereas the application of high pressure led to a noticeable decrease of color development in the carbonate-bicarbonate system. PMID- 12643649 TI - Chemical and sensory evaluation of crude oil extracted from herring byproducts from different processing operations. AB - Fish oils extracted from marinated herring (frozen and unfrozen) byproducts and maatjes herring byproducts were evaluated on their chemical and sensory properties. The obtained crude oils had very low content of copper (<0.1 mg/kg oil), and iron values were 0.8, 0.1, and 0.03 mg/kg oil, respectively, for oil from maatjes and frozen and fresh byproducts. For the maatjes oil, a much lower value was found for alpha-tocopherol compared to the other oils. Storage stability results showed that the oils behave differently. Secondary oxidation products were measured for fresh oil, while for the maatjes and frozen byproducts' oil, tertiary oxidation products were detected. Over storage time, the maatjes and frozen byproducts' oils became more intense in odor, correlating positively at the end with sensory attributes of train-oil, acidic, marine and fishy. The best correlation between sensory and chemical analyses was found for FFA and fishy off-odor (r = 0.781). PMID- 12643650 TI - Influence of cultivar, cooking, and storage on cell-wall polysaccharide composition of winter squash (Cucurbita maxima). AB - Changes in the cell-wall polysaccharides (CWP) of the edible tissues of four winter squash cultivars during storage and after cooking were investigated. A procedure for isolating cell walls of tissues containing high levels of starch was used. The starch-free CWP were sequentially fractionated using CDTA, dilute Na(2)CO(3), and 4 M KOH. Cellulose made up 40-42% of the total CWP for three cultivars (Delica, CF 2, and CF 4) at harvest but was 35% in the softer Red Warren. The pectic polysaccharides of Delica, CF 2, and CF 4 cell walls are more branched than those from Red Warren squash. The higher proportion of uronic acid in the pectic polysaccharides of Red Warren squash correlates with its lower firmness. Cooking resulted in an increase in the water-soluble pectins and a decrease in the pectins associated with cellulose. The total CWP content of the squash cultivars remained unchanged for up to 2 months of storage and then markedly decreased between 2 and 3 months of storage. The galactose content of Delica and Red Warren cell walls remained relatively constant from harvest to 2 months of storage and then decreased markedly during 2-3 months of storage. PMID- 12643651 TI - Barrier and mechanical properties of milk protein-based edible films containing nutraceuticals. AB - Calcium caseinate (CC) and whey protein isolate (WPI) films were prepared to contain 5 or 10% Gluconal Cal (GC), a mixture of calcium lactate and gluconate, or 0.1 or 0.2% alpha-tocopheryl acetate (VE), respectively. The pH and viscosity of film-forming solutions and the water vapor permeability and tensile property of the films were determined using standard procedures. CC and WPI films have the capabilities to carry high concentration of GC or VE, but some of the film functionality might be compromised. Adding VE to CC and WPI films increased film elongation at break, whereas incorporating 0.2% VE decreased WVP of CC films and tensile strength of both CC and WPI films. Incorporation of GC reduced the tensile strength of CC films (P < 0.05), with 10% GC decreasing both elongation at break and WVP (P < 0.05). These types of films may be used for wrapping or coating to enhance the nutritional value of foods. The concentration of GC and VE added to the films must be carefully selected to meet required water barrier and mechanical properties of the films depending on their specific applications. PMID- 12643652 TI - A new class of blue anthocyanin-derived pigments isolated from red wines. AB - Two newly formed anthocyanin-derived pigments that revealed unique spectroscopic features, showing maximum absorption in their UV-vis spectra at 575 nm, were isolated by TSK Toyopearl HW-40 (S) gel column chromatography and semipreparative HPLC from an aged Port red wine. Further characterization by ESI/MS and NMR ((1)H, gCOSY) showed them to belong to a new class of pigments described here for the first time, the structure of which consisted of a pyranoanthocyanin moiety linked to a flavanol by a vinyl bridge. The extended conjugation of the pi electrons throughout all the pigment molecule is likely to confer a higher stability on it and is probably the origin of the intense blue color. The formation of these pigments was found to arise from the reaction between anthocyanin-pyruvic acid adducts and vinyl-flavanol adducts. PMID- 12643653 TI - Effects of processing treatment and sorbate addition on the flavor characteristics of apple cider. AB - Processing treatments used to produce a microbiologically "safe" apple cider were evaluated to determine the impact of these treatments on the overall flavor characteristics. Apple cider with (0.1%) and without (0%) potassium sorbate was subjected to four processing treatments: untreated, irradiated at 2 kGy, irradiated at 4 kGy, and pasteurized. Volatile flavor compounds were isolated from the cider using solid-phase microextraction methods with gas chromatographic analysis. A trained descriptive analysis panel evaluated sensory attributes. The effects of the processing treatment were dependent on the presence of sorbate in the apple cider. Irradiation treatments resulted in a decrease in the content of esters characteristic of apple flavor and an increase in the content of alcohols and aldehydes formed through lipid oxidation reactions. The presence of sorbate reduced the effects of the irradiation treatments on these volatile flavor compounds. Sensory panelists, however, detected higher intensities of undesirable flavor attributes, including "cardboard flavor", and lower intensities of the desirable "apple flavor" in irradiated cider with added sorbate. PMID- 12643654 TI - Reaction mechanisms in delignification of pine Kraft-AQ pulp with hydrogen peroxide using Mn(IV)-Me4DTNE as catalyst. AB - Pine Kraft-AQ pulp was bleached with hydrogen peroxide catalyzed by [LMn(IV)(2) (mu-O)(3)](ClO(4))(2) at 80 degrees C for 120 min under optimum reaction conditions. The resulting bleached pulp was hydrolyzed with cellulase to obtain insoluble and soluble residual lignins. The alkaline effluent from the bleaching was acidified to precipitate alkaline soluble lignin. These lignin preparations were purified, and then analyzed by 2D HMQC NMR spectroscopic techniques. The results showed that biphenyl (5-5) and stilbene structures are preferentially degraded in the bleaching process, while beta-O-4, beta-5, and beta-beta structures undergo degradation only to a lesser extent. This implies that hydrogen peroxide bleaching using the catalyst is more effective in delignification of softwood pulps than hardwood pulps. The possible reaction mechanisms for the delignification of residual lignin in the pine Kraft-AQ pulp in the bleaching process are discussed on the basis of the 2D HMQC NMR spectroscopic data and the model compound experiments. PMID- 12643655 TI - Fatty acid distribution in muscle, liver, and gonads of rays (Dasyatis marmorata, Rhinobatos cemiculus, and Rhinoptera marginata) from the East Tropical Atlantic Ocean. AB - If a great number of rays are fished in the Tropical East Atlantic Ocean for their caudal fins, only a small amount of ray flesh is processed. Among them, three species of rays, Dasyatis marmorata, Rhinobatos cemiculus, and Rhinoptera marginata, from the Mauritanian coast have been investigated for the fatty acid composition of their lipids. Gas chromatography and gas chromatography-mass spectrometry allowed identification of 50 molecules from muscles, livers, and gonads of these fishes. Principal component analysis, starting from >50 samples, reveals significant differences in various fatty acid distributions, related to the species and sex of the sampled fish. Some of them are preferentially present in one sex or in both species, whereas the occurrence of others characterizes the male and female of one or two species. The results show that rays are potential resources of polyunsaturated fatty acids (PUFA) and should be used in the diet of local populations. The lipidic fractions contained a high amount of PUFA (up to 30% of the total), mainly composed of docosa-4,7,10,13,16,19-hexaenoic acid, eicosa-5,8,11,14-tetraenoic acid, and eicosa-5,8,11,14,17-pentaenoic acid. PMID- 12643657 TI - Antifeedant activity of some pentacyclic triterpene acids and their fatty acid ester analogues. AB - The 3-O-fatty acid ester derivatives (C(12)-C(18)) of two pentacyclic triterpenic acids, ursolic acid and oleanolic acid, were synthesized under mild esterification conditions in excellent yields (80-85%) and screened for their antifeedant activity, together with the parent acids, against the agricultural pest tobacco caterpillar larvae (Spodoptera litura F) in a no-choice laboratory study. The Urs-12-ene-28-carboxy-3beta-octadecanoate and olean-12-ene-28-carboxy 3beta-hexadecanoate were found to exhibit exceptionally potent antifeedant activities at 50 microg/cm(2) concentration, even after 48 h. PMID- 12643656 TI - Cyclooxygenase inhibitory and antioxidant compounds from crabapple fruits. AB - Crabapple trees belong to the Malus genus (Rosaceae) and bear fruits that are sparingly consumed and used in the preparation of fruit beverages. Cyclooxygenase (COX) enzyme inhibitory and antioxidant bioassay-guided fractionation of the aqueous and methanol extracts of Malus x kornicensis and Malus x Indian Summer yielded (+)-catechin (1), (-)-epicatechin (2), cyanidin-3-O-beta galactopyranoside (3), and amygdalin (4). Pure compounds 1-4 were obtained by HPLC, identified by LC-ES/MS, CD, and NMR spectroscopic methods and evaluated for their COX enzyme inhibitory and antioxidant activities. In COX-1 and -2 enzyme inhibitory assays, compounds 1-3 (all at 80 microM) showed activities of 20.4, 46.3%; 57.6, 47.9%; and 8.2, 13.7%, respectively, compared to naproxen (54.3, 41.3%; 10 microM), ibuprofen (47.5, 39.8%; 10 microM), Celebrex (46.2, 66.3%; 1.67 ppm), and Vioxx (23.8, 88.1%, 1.67 ppm). In the antioxidant assay, the catechins (1-2) and anthocyanin (3) (all at 40 microM) showed activities of 61.3, 62.5, and 60.1%, respectively. The synthetic antioxidants, tert-butylhydroquinone (TBHQ), butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), and vitamin E (all tested at 10 microM), gave 75.2, 80.1, 70.0, and 10.2% activities, respectively. The cyanogenic glycoside, amygdalin (4), and its hydrolysis products, mandelonitrile (5) and benzaldehyde (6), were not active in the antioxidant or COX enzyme inhibitory assays at 80 microM concentrations. PMID- 12643658 TI - Reaction of chlorpyrifos-methyl in aqueous hydrogen sulfide/bisulfide solutions. AB - The kinetics of the reactions of chlorpyrifos-methyl, an organophosphorus insecticide, with hydrogen sulfide (H(2)S) and bisulfide (HS(-)) were determined in well-defined aqueous solutions. The resulting pseudo-first-order rate constant for chlorpyrifos-methyl with bisulfide yielded a second-order rate constant of (2.1 +/- 0.3) x 10(-3) M(-1) s(-1). The second-order rate constant for chlorpyrifos-methyl with hydrogen sulfide is significantly slower than the second order rate constant with bisulfide. The contribution of H(2)S to the observed degradation rate constant of chlorpyrifos-methyl at concentrations of up to 4 mM H(2)S is not significant. The second-order rate constant of chlorpyrifos-methyl with H(2)S was too low to be measured in this study. The results indicate that HS(-) present at environmentally relevant concentrations may represent an important sink for phosphorothionate triesters in a coastal marine environment, while H(2)S reacts too slowly to be environmentally relevant (pH 6-9). Trichloropyridinol, the major product of hydrolysis of chlorpyrifos-methyl, is only a minor product of the reaction of chlorpyrifos-methyl with bisulfide; however, trichloropyridinol was found to be stable under the experimental conditions. PMID- 12643659 TI - Analysis of volatile compounds released during the grinding of roasted coffee beans using solid-phase microextraction. AB - A dynamic solid-phase microextraction (SPME) method to sample fresh headspace volatile compounds released during the grinding of roasted coffee beans was described and the analytical results using gas chromatography/mass spectrometry (GC/MS) and GC/olfactometry (GC/O) were compared to those of the conventional static SPME sampling methods using ground coffee. Volatile compounds released during the grinding of roasted coffee beans (150 g) were obtained by exposing the SPME fiber (poly(dimethylsiloxane)/divinylbenzene, PDMS/ DVB) for 8 min to nitrogen gas (600 mL/min) discharged from a glass vessel in which the electronic coffee grinder was enclosed. Identification and characterization of volatile compounds thus obtained were achieved by GC/MS and GC/O. Peak areas of 47 typical coffee volatile compounds, separated on total ion chromatogram (TIC), obtained by the dynamic SPME method, showed coefficients of variation less than 5% (n = 3) and the gas chromatographic profile of volatile compounds thus obtained was similar to that of the solvent extract of ground coffee, except for highly volatile compounds such as 4-hydroxy-2,5-dimethyl-3(2H)-furanone and 4-ethenyl-2 methoxyphenol. Also, SPME dilution analysis of volatile compounds released during the grinding of roasted coffee beans showed linear plots of peak area versus exposed fiber length (R (2) > 0.89). Compared with those of the headspace volatile compounds of ground coffee using GC/MS and GC/O, the volatile compounds generated during the grinding of roasted coffee beans were rich in nutty- and smoke-roast aromas. PMID- 12643660 TI - Formation of aroma compounds and lipoxygenase (EC 1.13.11.12) activity in unblanched leek (Allium ampeloprasum Var. Bulga) slices during long-term frozen storage. AB - The content of aroma compounds (dynamic headspace) and catalytic activity of lipoxygenase (LOX) (EC. 1.13.11.12) were analyzed in 15 mm unblanched leek slices seven times during 12 months of frozen storage. The aroma profile changed from consisting of almost only sulfur compounds such as dipropyl disulfide [concentration in fresh leek (FL) = 0.197 mg/L, concentration after 12 months of frozen storage (12M) = 0.0409 mg/L] and propyl (E)-propenyl disulfide (FL = 0.0437 mg/L, 12M = 0.00452 mg/L) in the fresh leeks to being dominated by numerous saturated and unsaturated aldehydes, such as hexanal (FL = 1.53 mg/L, 12M = 3.63 mg/L), (E,E)-2,4-nonadienal (FL = 0.000 mg/L, 12M = 0.0647 mg/L), and (E,E)-2,4-decadienal (FL = 0.129 mg/L, 12M = 0.594 mg/L) at the end of the storage period. The catalytic activity of LOX diminished throughout frozen storage, but approximately 25% of the original activity was present after 12 months of storage. PMID- 12643661 TI - Use of catalyst in a 3D-QSAR study of the interactions between flavor compounds and beta-lactoglobulin. AB - This paper reports a 3D-QSAR study using Catalyst software to explain the nature of interactions between flavor compounds and beta-lactoglobulin. A set of 35 compounds, for which dissociation constants were previously determined by affinity chromatography, was chosen. The set was divided into three subsets. An automated hypothesis generation, using HypoGen software, produced a model that made a valuable estimation of affinity and provided an explanation for the lack of correlation previously observed between the hydrophobicity of terpenes and the affinity for the protein. On the basis of these results, it appears that aroma binding to beta-lactoglobulin is caused by both hydrophobic interactions and hydrogen bonding, which plays a critical role. Catalyst appears to be a reliable tool for the application of 3D-QSAR study in aroma research. PMID- 12643662 TI - Effect of cis/trans isomerism of beta-carotene on the ratios of volatile compounds produced during oxidative degradation. AB - beta-Carotene is, when cleaved, an important source of flavor and aroma compounds in fruits and flowers. Among these aroma compounds, the main degradation products are beta-ionone, 5,6-epoxy-beta-ionone, and dihydroactinidiolide (DHA), which are associated by flavorists and perfumers with fruity, floral, and woody notes. These three species can be produced by degradation of beta-carotene through an attack by enzyme-generated free radicals and a cleavage at the C9-C10 bond. This study investigated the influence of cis/trans isomerism at the C9-C10 bond on the production of beta-carotene degradation compounds, first with a predictive approach and then experimentally with different isomer mixtures. beta-Carotene solutions containing high ratios of 9-cis-isomers produced more DHA, suggesting a different pathway than for the transformation of all-trans-beta-carotene to ionone and DHA. These results are important in the search for financially viable processes to produce natural carotene-derived aroma compounds. PMID- 12643663 TI - Investigation of binding behavior of alpha- and beta-ionones to beta lactoglobulin at different pH values using a diffusion-based NOE pumping technique. AB - Diffusion-based NMR techniques were employed to study effects of pH on beta lactoglobulin (BLG) conformation and binding affinity to alpha- and beta-ionone. In the first part of the study, the influence of pH on the diffusion coefficient of BLG in D(2)O solution was investigated using a stimulated-echo NMR experiment. The diffusion coefficient of BLG decreased with increasing pH values. A significant decrease in the diffusion coefficient observed at pH 11 may be due to total unfolding (denaturation) of the protein, resulting in hydrophobically driven self-aggregation. A diffusion-based NOE pumping technique was then applied to determine the relative binding affinities between alpha- and beta-ionones and BLG at pH values varying from 3 to 11. An increase in signal intensities for beta ionone with increasing molar concentration ratios between beta-ionone and BLG was observed at all pH ranges studied. The increased signal intensities reflect increased relative binding affinity. The greatest binding affinity occurred at pH 9 and the lowest at pH 11. alpha-Ionone showed binding evidence only at pH 9, and the binding was significantly weaker than that obtained for beta-ionone at the same pH. The high affinity observed for both aroma compounds at pH 9 may be due to a flexible conformation of BLG at this pH so that the flavor ligand accessibility increases. Conversely, alkaline denaturation occurring at pH 11 gives rise to relatively lower binding affinity compared to that observed at the other pH values. PMID- 12643664 TI - Volatile fractions from three cultivars of Olea europaea L. collected in two different seasons. AB - The chemical composition of the volatile fractions from leaves of three Olea europaea L. cultivars (Leccino, Frantoio, and Cipressino) harvested at two different times of the year were examined by GC and GC-MS. The results showed a high content of aliphatic aldehydes in the three cultivars during both harvesting periods and an increase of (E)-2-hexenal (an aldehyde with high antimicrobial properties) percentage from July to November. PMID- 12643665 TI - Partition and release of 21 aroma compounds during storage of a pectin gel system. AB - The increasing popularity of low-fat products increases the need for a better understanding of how flavor release is affected by partial substitution of fat with hydrocolloids. Partitioning and release of aroma compounds from four pectin gels with different compositions were studied with static headspace and with a model mouth. Air/product partition coefficients determine the potential extent of aroma release, and mass transfer determines the rate at which aroma compounds are released to the vapor phase. This study showed that the gel network had large effects on the partition of aroma compounds between the gel and vapor phase. The specific properties of the aroma compounds were also of importance for the air/gel partition. Storage of the four gels showed that one of the weaker gels was influencing the concentration of aroma compounds in the headspace, probably caused by formation of a denser network over time. PMID- 12643666 TI - Comparison of two microalgal diets. 1. Influence on the biochemical and fatty acid compositions of raw oysters (Crassostrea gigas). AB - Oyster farming in France is a traditional activity. Each year, 149 000 tons of oysters are fattened before being sold. More and more oyster farmers supplement the diet of oysters by microalgae to optimize the fattening process and to improve both the growth and flesh quality of oysters. In the present study, oysters were supplemented by two microalgae: Skeletonema costatum and Tahitian isochrysis clone. The ash, protein, carbohydrate (including glycogen), and lipid contents were analyzed. The growth of oysters was improved when they were fattened with microalgae, leading to an increase in the condition index. The chemical composition of oysters was influenced by the chemical composition of the microalgae, especially the level of glycogen, which was significantly increased for oysters supplemented by microalgae. The fatty acid profile of oysters fattened by microalgae was positively correlated with the fatty acid profiles of S. costatum and T. isochrysis. These results show the effectiveness of supplementation by microalgae on the growth and on the biochemical composition (glycogen and fatty acids especially) of oysters. PMID- 12643667 TI - Comparison of two microalgal diets. 2. Influence on odorant composition and organoleptic qualities of raw oysters (Crassostrea gigas). AB - Oyster farming is of real economic interest in France. Oyster farmers attach more and more importance to improving the growth and the quality of their oysters. Some fatty acids known to be aroma precursors originate from microalgae such as Skeletonema costatum and Tahitian isochrysis clone. These microalgae were used to fatten oysters in order to observe their role in the development of oysters' aroma. This study shows that the profile of fatty acids of oysters is influenced by the contribution of fatty acids from the two microalgae (as reported in the first paper in this series: Pennarun, A.-L.; Prost, C.; Haure, J.; Demaimay, M. Comparison of Two Microalgal Diets. 1. Influence on the Biochemical and Fatty Acid Compositions of Raw Oysters (Crassostrea gigas). J. Agric. Food Chem. 2003, 51, 2006-2010 (in this issue)]. As a consequence, a microalgal diet causes changes in oysters' aroma composition. Aroma concentration depends on the content of fatty acids that are aroma precursors in oysters. Some aromas are characteristic of the diet of S. costatum, such as 6-methyl-5-hepten-2-one (ether odor), and others are characteristic of T. isochrysis, such as 3-nonyne (cucumber, marine odor), 6-(E)-nonen-1-ol (green and fresh odor), and 4 ethylbenzaldehyde (aniseed odor). Moreover, the organoleptic qualities (odor, taste, and texture) of oysters are modified by the diet of microalgae. PMID- 12643668 TI - Identification of the wheat seed protein CM3 as a highly active emulsifier using a novel functional screen. AB - Lyophilized albumin protein fractions were prepared from flour of four varieties of wheat: Triticum aestivum cvs. Mercia and Riband, Triticum aestivum var. spelta, and Triticum turgidum var. durum (Kamut). The dry powders were redissolved in sodium phosphate buffers at pH 3.0, 6.5, or 8.0 and at ionic strengths of 0.1 or 1.0 M to a concentration of 0.1% (w/v). Emulsions formed by sonication of protein solutions with n-hexadecane were aged at room temperature and separated into aqueous, interstitial, and interfacial phases. The distinct emulsion components were lyophilized and analyzed by RP-HPLC. A protein was observed to be preferentially located in the interfacial component and subsequently purified from a total albumin fraction and identified by N-terminal sequencing as CM3, an alpha-amylase inhibitor subunit. Measurement of the equilibrium surface tension of CM3 as a function of protein concentration demonstrated that it was at least as active as bovine beta-lactoglobulin, an established protein emulsifier. Furthermore, measurement of the surface dilational elastic modulus at an air/water interface demonstrated the formation of a viscoelastic film, while fluorescence and FT-IR spectroscopic measurements on adsorbed and nonadsorbed CM3 suggest that the secondary structure is essentially unchanged upon adsorption to an oil/water interface. It is concluded that functional screening is a valid approach to identify novel protein emulsifiers in complex mixtures. PMID- 12643669 TI - Individual contribution of grain outer layers and their cell wall structure to the mechanical properties of wheat bran. AB - The mechanical properties of wheat bran and the contribution of each constitutive tissue on overall bran properties were determined on a hard wheat (cv. Baroudeur) and a soft wheat (cv. Scipion). Manual dissection allowed three different layers to be separated from wheat bran, according to radial and longitudinal grain orientations, which were identified by confocal laser scanning microscopy as outer pericarp, an intermediate strip (comprising inner pericarp, testa, and nucellar tissue), and aleurone layer, respectively. Tissue microstructure and cell wall composition were determined. Submitted to traction tests, whole bran, intermediate, and aleurone layers demonstrated elastoplastic behavior, whereas pericarp exhibited elastic behavior. By longitudinal orientation, pericarp governed 50% bran elasticity (elastic strength and rigidity), whereas, in the opposite orientation, bran elastic properties were mostly influenced by the other tissues. Regardless of test orientation, the linear force required to bran rupture corresponded to the sum of intermediate and aleurone layer strengths. According to radial orientation, the intermediate strip governed bran extensibility, but according to longitudinal orientation, all tissues contributed until bran disruption. Tissues from both wheat cultivars behaved similarly. A structural model of wheat bran layers illustrated the detachment of pericarp from intermediate layer within radial bran strips. PMID- 12643670 TI - Thermal stability of texture in Chinese water chestnut may be dependent on 8,8' diferulic acid (aryltetralyn form). AB - Ferulic acid (FA) cross-links have been implicated in the thermal stability of texture in Chinese water chestnut (CWC) tissues. The aim of the current study has been to investigate this concept further. CWC tissue strips were measured for their mechanical properties before and after extraction in increasing strengths of alkali. The mechanical properties were related to the associated mode of fracture (cell separation or breakage) at the fracture surfaces and the phenolic composition of the cell walls. CWC tissue softened after prolonged extraction in cold alkali due to an increase in the ease of cell separation. Analysis of wall bound phenolics demonstrated that most FA moieties, including five of the six dehydrodimers, were released before tissue strength was reduced. Loss of strength was, however, coincident with the loss of 8,8'-diferulic acid, aryltetralin (AT) form. It has been suggested that this dehydrodimer may be particularly concentrated at the edge of the cell faces. These results provide further evidence for the involvement of this dehydrodimer in conferring thermal stability of cell-cell adhesion and hence texture in CWC. However, they do not exclude the other diferulates from involvement in cell adhesion. PMID- 12643671 TI - Clarification of Muscat musts using wheat proteins and the flotation technique. AB - The bovine spongiform encephalopathy (BSE, mad cow) crisis has led some wine makers to question gelatin as a fining agent and to reject the use of these animal proteins. The search for a substitute for gelatin was begun by comparing vegetable proteins (particularly gluten) with gelatin fining treatments. Clairette de Die (French-controlled appellation) is a sparkling sweet wine. The alcoholic fermentation begins in a tank, continues in a crown-cap bottle, and stops before the complete consumption of sugar. All particles present in the bottle have to be removed before corking, and it is important to have a must as clear as possible. This study concerned the clarifying of a Muscat must, treated with pectinases, using a very efficient flotation technique. The must is fined with bentonite/silica/protein (fish gelatin, wheat gluten, or lupin isolate) to induce flocculation and then pressurized (6 bar). After depressurization, microbubbles cling to flocculates and climb to the top of the flotation tank (this flotation foam is clarified using a rotary filter). At laboratory scale, gluten (20 g/hL) and gelatin (10 g/hL) (each combined with bentonite and silica gel) gave turbidities of 50 and 35 NTU, respectively (6.5 and 4.1% of that of the nontreated must). The Muscat must was also clarified by static settling. The turbidity decreased by 86% for the gluten/bentonite fining and by 60% for the gelatin/bentonite fining. Visually, gluten flocculation takes longer to occur and flocculates sedimentation is longer than with gelati, but the removal of insoluble particles is more complete and leads to lower turbidities. At an industrial scale, gluten (20 g/hL) and gelatin (10 g/hL) (each combined with bentonite and silica gel) gave turbidities of 60 and 48 NTU, respectively. Turbidities measured in the tanks 14 h after the flotation showed a better efficiency for the wheat gluten (24 NTU) compared to gelatin (28 NTU). This is explained by the static settling that completed the clarifying effect of the flotation. The last experiment showed comparable efficiencies for wheat gluten and lupin proteins. BSE caused a situation of crisis (in Europe particularly) leading the public and wine-makers to lose their confidence in the use of gelatin as fining agent and to reject animal proteins in general. It is proposed here that vegetable proteins could efficiently replace gelatin. PMID- 12643672 TI - Charge state distribution and hydrogen/deuterium exchange of alpha-lactalbumin and beta-lactoglobulin preparations by electrospray ionization mass spectrometry. AB - Charge state distribution (CSD) and hydrogen/deuterium (H/D) exchange of preparations of alpha-lactalbumin (alpha-Lac) and beta-lactoglobulin (beta-Lg) were investigated using electrospray ionization mass spectrometry (ESI-MS). Storage of alpha-Lac at pH 3 resulted in substantial changes in its CSD, with the emergence of new ion species and shifts toward higher charge state, indicating less stable conformation. ESI spectra of alpha-Lac kept at pH 5.5 for 4 days showed stable conformation; however, extending the storage period resulted in substantial changes in CSD and a decrease in the stability of holo-alpha-Lac (Ca(2+)-bound form). In comparison to apo-alpha-Lac, the relative intensity of holo-alpha-Lac was higher at pH 6.8 but lower at pH 8 during the storage period. beta-Lg showed stable CSD at pH 3, substantial changes at pH 5.5, and minor changes at pH 6.8 and 8 during storage. The H/D exchange results demonstrate that the conformation of holo-alpha-Lac was more stable than that of apo-alpha-Lac and that the conformation of beta-Lg variant B was more stable than that of the beta Lg variant A. Kinetics of H/D exchange indicated that alpha-Lac and beta-Lg fractions obtained from whey protein preparations have the same or improved conformational stabilities compared to those of alpha-Lac and beta-Lg standards. The presence of four or more hexose residues in alpha-Lac enhanced its conformational stability; the presence of two hexose residues in beta-Lg resulted in a less stable conformation. PMID- 12643674 TI - Post-mortem changes in porcine M. longissimus studied by solid-state 13C cross polarization magic-angle spinning nuclear magnetic resonance spectroscopy. AB - Solid-state (13)C cross-polarization (CP) magic-angle spinning (MAS) nuclear magnetic resonance (NMR) experiments are carried out for the first time on rapidly frozen muscle biopsies taken in M. longissimus in vivo and at 1 min, 45 min, and 24 h post-mortem from three pigs. Two of the pigs were CO(2)-stunned (control animals), and one was pre-slaughter-stressed (treadmill exercise) followed by electrical stunning to induce difference in metabolism post-mortem. (13)C resonance signals from saturated and unsaturated carbons in fatty acids, carboxylic carbons, and carbons in lactate and glycogen are identified in the solid-state NMR spectra. The (13)C CP MAS spectra obtained for post-mortem samples of the stressed, electrically stunned pig differ significantly from the post-mortem control samples, as the intensity of a resonance line appearing at 30 ppm, assigned to carbons of the methylene chains, is reduced for the stressed pig. This spectral difference is probably due to changes in lipid mobility and indicates altered membrane properties in the muscle of the stressed/electrically stunned animal when compared with the control animals already 1 min post-mortem. In addition, the post-mortem period changes in glycogen carbons can be estimated from the (13)C CP MAS spectra, yielding a correlation of r = 0.74 to subsequent biochemical determination of the glycogen content. PMID- 12643673 TI - Partial purification of latent persimmon fruit polyphenol oxidase. AB - Persimmon fruit polyphenol oxidase (PPO) was partially purified using a combination of phase partitioning with Triton X-114 and ammonium sulfate fractionation between 50 and 75%. The enzyme, which showed both monophenolase and diphenolase activities, was partially purified in a latent form and could be optimally activated by the presence of 1 mM sodium dodecyl sulfate (SDS) with an optimum pH of 5.5. In the absence of SDS, the enzyme showed maximum activity at acid pH. SDS-PAGE showed the presence of a single band when L-DOPA was used as substrate. The apparent kinetic parameters of the latent enzyme were determined at pH 5.5, the V(m) value being 15 times higher in the presence of SDS than in its absence, whereas the K(M) was the same in both cases, with a value of 0.68 mM. The effect of several inhibitors was studied, tropolone being the most active with a K(i) value of 0.45 microM. In addition, the effect of cyclodextrins (CDs) was studied, and the complexation constant (K(c)) between 4-tert-butylcatechol (TBC) and CDs was calculated using an enzymatic method. The value obtained for K(c) was 15580 M(-1). PMID- 12643676 TI - Natural abundance hydrogen isotope affiliation between the reactants and the products in glucose fermentation with yeast. AB - In glucose fermentation, the hydrogen source of products such as ethanol and glycerol is the medium and the sugar. The site-specific natural isotope ratios of the products, (D/H)(i), and that of the medium and sugar, (D/H)(k), may be related by a matrix, A, of redistribution coefficients, a(ik), that characterizes the specific genealogies of the hydrogen atoms. (D/H)(i) = [A](D/H)(k), where (D/H)(i) and (D/H)(k) are the column vectors of the isotope ratios of sites i and k that can be measured by (2)H NMR. The complete redistribution matrix was determined in a set of isotope labeling experiments. Thus, we obtained a mathematical model representing the hydrogen isotope affiliation during alcoholic fermentation. It not only provides information about the biochemical reaction mechanism but also can be used to estimate the isotopic data of the products, based on those of the substrate and the medium. The results prove, in a quantitative way, that the metabolites contain isotopic information about the precursor in a biotransformation and can be used to identify its origin. The method established for the study of the hydrogen-transfer mechanism can be applied to other chemical and biochemical reactions. PMID- 12643677 TI - Antioxidative and antiplatelet effects of aqueous inflorescence Piper betle extract. AB - Piper betle, belonging to the Piperaceae family, is a tropical plant, and its leaf and inflorescence are popularly consumed by betel quid (BQ) chewers in Taiwan and many other South and Southeast Asian countries. However, little is known about the biochemical properties of inflorescence Piper betle (IPB) toward reactive oxygen species (ROS) and platelet functions. In the present work, aqueous IPB extract was shown to be a scavenger of H(2)O(2), superoxide radical, and hydroxyl radical with a 50% inhibitory concentration (IC(50)) of about 80, 28, and 73 microg/mL, respectively. IPB extract also prevented the hydroxyl radical induced PUC18 plasmid DNA breaks at concentrations higher than 40 microg/mL. Since ROS are crucial for platelet aggregation, we further found that IPB extract also inhibited the arachidonic acid (AA) induced and collagen-induced platelet aggregation, with an IC(50) of 207 and 335 microg/mL, respectively. IPB extract also inhibited the AA-, collagen- (>100 microg/mL of IPB), and thrombin (>250 microg/mL of IPB)-induced thromboxane B(2) (TXB(2)) production by more than 90%. However, IPB extract showed little effect on thrombin-induced aggregation. These results indicated that aqueous components of IPB are potential ROS scavengers and may prevent the platelet aggregation possibly via scavenging ROS or inhibition of TXB(2) production. PMID- 12643675 TI - Separation and characterization of a salt-dependent pectin methylesterase from Citrus sinensis var. Valencia fruit tissue. AB - A pectin methylesterase (PME) from sweet orange fruit rag tissue, which does not destabilize citrus juice cloud, has been characterized. It is a salt-dependent PME (type II) and exhibits optimal activity between 0.1 and 0.2 M NaCl at pH 7.5. The pH optimum shifted to a more alkaline range as the salt molarity decreased (pH 8.5-9.5 at 50 mM NaCl). It has an apparent molecular mass of 32.4 kDa as determined by gel filtration chromatography, an apparent molecular mass of 33.5 kDa as determined by denaturing electrophoresis, and a pI of 10.1 and exhibits a single activity band after isoelectric focusing (IEF). It has a K(m) of 0.0487 mg/mL and a V(max) of 4.2378 nkat/mg of protein on 59% DE citrus pectin. Deblocking the N-terminus revealed a partial peptide composed of SVTPNV. De esterification of non-calcium-sensitive pectin by 6.5% increased the calcium sensitive pectin ratio (CSPR) from 0.045 +/- 0.011 to 0.829 +/- 0.033 but had little, if any, effect on pectin molecular weight. These properties indicate this enzyme will be useful for studying the PME mode of action as it relates to juice cloud destabilization. PMID- 12643678 TI - Study of low molecular weight phenolic compounds during the aging of sparkling wines manufactured with red and white grape varieties. AB - Thirty-two phenolic compounds of low molecular weight were identified in 36 white, blanc de noir, and rose sparkling wines by using HPLC with photodiode array and mass spectrometry detection. Some of the identified compounds, such as cis- and trans-ethylcaftaric, cis- and trans-ethylcaffeic, and cis- or trans ethyl-p-coumaric acids, 2R,3R-dihydroquercetin, 2R,3R-dihydrokaempferol 3-O-beta d-glucoside, and a lignan derivative are described for the first time in sparkling wines manufactured with grapes of red varieties. This is also the first time that cis- or trans-diethylfertaric acids have been identified in wines. When cluster analysis was applied to the data of 19 of the 32 identified compounds, the greatest differences found in the low molecular weight phenolic compounds in sparkling wines were due to the grape variety from which they were manufactured, whereas aging time did not significantly influence phenolic composition. Nine phenolic compounds, that is, trans-p-coumaric and trans-caftaric acids, trans resveratrol glucoside, cis-coutaric, trans-coutaric, cis-p-coumaric, and cis caftaric acids, tryptophol, and syringic acid, permit the wines to be classified correctly in accordance with the grape variety from which they were manufactured. PMID- 12643679 TI - Synthesis and analysis of symmetrical and nonsymmetrical disaturated/monounsaturated triacylglycerols. AB - Symmetrical disaturated triacylglycerols of the structure SUS, where S is stearic acid (18:0) and U is an unsaturated fatty acid, either oleic (O; 9cis-18:1), linoleic (L; 9cis,12cis-18:2), or linolenic (Ln; 9cis,12cis,15cis-18:3), are important components providing functionality to interesterified fat blends and structurally modified oils. Nonsymmetrical triacylglycerols of the structure SSU can significantly change melting point and solid fat content profiles. To characterize the physical properties of pure and symmetrical and nonsymmetrical triacylglycerol mixtures, the same reaction sequence has been used to prepare multigram quantities of triacylglycerols SUS and SSU. Tristearin was converted to a mixture of mono-, di-, and triacylglycerols, and the 1,3- and 1,2 diacylglycerol fraction was isolated by silica column chromatography. The 1,3 diacylglycerols were removed by crystallization from acetone and esterified with the appropriate fatty acid to form the symmetrical triacylglycerols with >99% SUS structure. The more difficult to obtain 1,2-diacylglycerols were prepared by esterification of the enriched 1,2-diacylglycerol fraction (80-86% 1,2 diacylglycerols) remaining after removal of much of the 1,3-isomer by crystallization, but silver resin or silver nitrate impregnated silica gel chromatography was required to isolate the nonsymmetrical triacylglycerols. SSL and SSLn were prepared in purities of >98% by this procedure, but not SSO. Silver ion HPLC was found to be as accurate as, and more rapid than, lipolysis/gas chromatography for the determination of the isomeric purities of the synthesized triacylglycerols. PMID- 12643680 TI - Characterization of an aspartic proteinase activity in buckwheat (Fagopyrum esculentum Moench) seeds. AB - The pepstatin A sensitive acidic proteolytic activity of total protein extracts of buckwheat seeds has been analyzed in developing, mature, and germinating seeds by activity measurements as well as by electrophoretic and immunochemical techniques. Immunoblot analysis using cross-reactive antibodies raised against barley phytepsin suggested that specific proteolytic activity could be attributed to a 47 kDa heterodimeric polypeptide, composed of two subunits: 31 and 16 kDa polypeptides. The analysis of time course expression revealed that the 47 kDa heterodimer accumulated during seed maturation starting from 12 days after pollination and was also present at the beginning of germination. Milk-clotting activity of this proteinase was also indicated. PMID- 12643681 TI - Synthesis of monoglucosylated high-mannose-type dodecasaccharide, a putative ligand for molecular chaperone, calnexin, and calreticulin. AB - Convergent and stereoselective synthetic routes to Man9GlcNAc2 (1b), alpha Glc1M9GlcNAc2 (2b), and its stereoisomer beta-Glc1M9GlcNAc2 (3) were established. Interaction analysis of 2b with CRT was measured by 1H NMR spectroscopy, and the first NMR-based evidence for the specific binding of CRT to 2b was obtained. PMID- 12643682 TI - Electrochromism in photochromic dithienylcyclopentenes. AB - Both photochromic and unprecedented electrochromic properties have been observed for a class of 1,2-bis(3-thienyl)cyclopentene derivatives bearing either a thiophene or a phenyl group attached to the reactive carbon centers. These compounds can be interconverted between their colorless ring-open and colored ring-closed states using UV and visible light, respectively. They can also be catalytically transformed from their ring-closed states to their ring-open states through electrochemical or chemical oxidation. PMID- 12643683 TI - Epoxidation of olefins by hydroperoxo-ferric cytochrome P450. AB - The T252A mutant of cytochrome P450cam is unable to form the oxoferryl "active oxygen" intermediate, as judged by its inability to hydroxylate its normal substrate, camphor. In the present study, we demonstrate that T252A P450cam is nonetheless able to epoxidize olefins, due to the action of a second oxidant. However, as shown in earlier radiolytic studies and by the ability of T252A to reduce dioxygen to hydrogen peroxide, the mutant retains the ability to form the hydroperoxo-ferric reaction cycle intermediate. The present results provide strong evidence that hydroperoxo-ferric P450 can serve as a second electrophilic oxidant capable of olefin epoxidation. PMID- 12643684 TI - Synthesis and optical properties of colloidal tungsten oxide nanorods. AB - Thermal decomposition of W(CO)6 in oleylamine in the presence of mild oxidant Me3NO.2H2O produces tungsten oxide nanorods with diameters ranging from 3 to 6 nm. The size of nanorods can be easily varied by the employed surfactant ratio or reaction temperature. The prepared tungsten oxide nanorods exhibit strong photoluminescence (PL) peaks in 300-500 nm, which show a weak size dependency. PMID- 12643685 TI - The complex of a designer antibiotic with a model aminoacyl site of the 30S ribosomal subunit revealed by X-ray crystallography. AB - The ribosome is an important target for aminoglycoside antibiotics; however, the clinical effectiveness of aminoglycosides has diminished due to bacterial resistance mechanisms. Here we report the X-ray structure of a novel synthetic aminoglycoside bound to the A site of the ribosome, its target for manifestation of activity. The structure validates the in silico design paradigms for the antibiotic and reveals the molecular interactions made by this novel antibiotic in prokaryotes. PMID- 12643686 TI - The polymer incarcerated method for the preparation of highly active heterogeneous palladium catalysts. AB - A new method named "the polymer incarcerated (PI) method" for preparing a heterogeneous palladium catalyst has been developed. The method is operationally simple, and the Pd catalyst prepared (PI Pd) is highly active for hydrogenation, carbon-carbon, and carbon-oxygen bond-forming reactions. Remarkable points are that the activity of PI Pd is higher than that of homogeneous Pd catalysts and that PI Pd is recovered by simple filtration and reused several times without loss of activity. The catalyst is expected to replace many heterogeneous palladium catalysts, especially Pd/C, which is often used in academia and industry, but recovery of which is difficult. PMID- 12643687 TI - The first stable beta-fluorosilylanion. AB - The reaction of 2 equiv of tris(trimethylsilyl)silylfluoride with potassium tert butoxide in the presence of donor molecules (THF, DME, 18-crown-6) leads to the clean formation of an adduct of 1-potassio-2 fluorotetrakis(trimethylsilyl)disilane. Attempts to transmetalate this compound effect the elimination of metal fluoride accompanied by the formation of tetrakis(trimethylsilyl)disilene. The latter can either be trapped in a cycloaddition reaction or in the absence of trapping reagents dimerizes to octakis(trimethylsilyl)cyclotetrasilane. PMID- 12643688 TI - Miniature homeodomains: high specificity without an N-terminal arm. AB - Recently, we described a strategy for the design of miniature proteins that bind DNA and protein surfaces with high affinity and selectivity. This strategy involves identifying the functional epitope required for macromolecular recognition by a natural protein and presenting it on a small, stable protein scaffold. In previous work, high-affinity DNA recognition was achieved only when the miniature protein contained the complete functional epitope. Here we report a miniature homeodomain that recognizes its 6-bp target site in the nanomolar concentration range at 25 degrees C, despite the absence of DNA contact residues located along the homeodomain N-terminal arm. We conclude that miniature proteins can achieve high affinity and selectivity for DNA by design even when the functional epitope is incomplete by using pre-organization to effectively compensate for lost protein-DNA contacts. In this case it has been possible to miniaturize both the recognition surface and the structural framework of a globular protein fold. PMID- 12643689 TI - Coupling selectivity with sensitivity in an integrated chemosensor framework: design of a Hg(2+)-responsive probe, operating above 500 nm. AB - For the highly selective and sensitive sensing of Hg2+ in water, a new design concept was realized where the selectivity of the probe's binding site is amplified by electronic properties of the chromophore. The molecular architecture of this phenoxazinone-type sensor molecule combines two potential coordination sites via an amino-keto conjugative backbone. These structural prerequisites allow only the most preferred mercuric ion to bind to the dithia dioxa monoaza crown unit, while other heavy, transition, and main group metal ions as well as protons are trapped at the keto group, inducing opposite spectral effects due to interaction with either the donor (Hg2+) or the acceptor group (other cations) of the probe. Besides these advantageous features, the probe operates well within the visible range of the spectrum and displays rather intense molar absorptivities as well as fluorescence quantum yields. PMID- 12643690 TI - Fluorescent detection of chemical warfare agents: functional group specific ratiometric chemosensors. AB - Indicators providing highly sensitive and functional group specific fluorescent response to diisopropyl fluorophosphate (DFP, a nerve gas (G-agent) simulant) are reported. Nonemissive indicator 2 reacts with DFP to give a cyclized compound 2+A that shows a high emission due to its highly planar and rigid structure. Very weak emission was observed by the addition of HCl. Another indicator based on pyridyl naphthalene exhibits a large shift in its emission spectrum after reaction with DFP, which provides for quantitative ratiometric detection. PMID- 12643691 TI - Catalytic transamidation under moderate conditions. AB - The carboxamide group is generally inert, except under harsh conditions or in the presence of highly evolved enzymes. We have identified several metal complexes that efficiently catalyze transamidation reactions of amide/amine mixtures under moderate conditions. This unprecedented reactivity represents an important step toward our long-term goal of using dynamic combinatorial chemistry to generate new amide-based molecules with interesting structures and functions. PMID- 12643692 TI - Cross-linked dendrimer hosts containing reporter groups for amine guests. AB - The integration of a chromogenic reporter group into the recently reported (Zimmerman, S. C.; Wendland, M. S.; Rakow, N. A.; Zharov, I.; Suslick, K. S. Nature 2002, 418, 399-403) monomolecular imprinting approach is described. The resulting highly cross-linked, macromolecular hosts show rapid, selective, high affinity, two-point binding of straight-chain diamine guests. Over longer times, the hosts are more promiscuous, binding a broader range of diamines. A rigorous test of imprinting was performed wherein the cross-reactivities of two dendrimers derived from different templates are compared. The test reveals a guest-dependent kinetic binding effect masquerading as evidence of a highly selective two-point imprinting process. PMID- 12643693 TI - Regio- and enantioselective iridium-catalyzed intermolecular allylic etherification of achiral allylic carbonates with phenoxides. AB - An enantioselective and regioselective iridium-catalyzed allylic etherification is described. The reaction of sodium and lithium aryloxides with achiral (E) cinnamyl and terminal aliphatic allylic electrophiles in the presence of 2 mol % of an iridium-phosphoramidite complex provides chiral allylic aryl ethers in high yields and excellent levels of regio- and enantioselectivity. Lithium aryloxides containing a single substituent at an ortho, meta, or para position as well as sterically hindered phenoxides were tolerated. Reactions in THF displayed the most suitable balance of rate, regio-, and enantioselectivity. High ee's were also observed for the products from the reaction of alkyl (E)-allylic carbonates. PMID- 12643695 TI - A new highly asymmetric chelation-controlled heck arylation. AB - This communication describes the development of a new highly asymmetric chelation controlled Heck arylation. The methodology permits formation of 2-aryl-2-methyl cyclopentanones in good to very good two-step yields (45-78%) with excellent chiral enrichment (90-98% ee). In contrast to the well-known direct coupling to the enolate of a cyclic ketone, the Heck arylation of the corresponding N-methyl pyrrolidin enol ether requires neither a strong base nor a blocking of the alternative nonalkylated alpha-carbon. A chelated pi-intermediate is proposed to explain the excellent chiral induction. PMID- 12643694 TI - Epothilone C macrolactonization and hydrolysis are catalyzed by the isolated thioesterase domain of epothilone polyketide synthase. AB - Epothilone C is produced by the combined action of one nonribosomal peptide synthetase (NRPS) and nine polyketide synthase (PKS) modules in a multienzyme system. The final step in the biosynthesis is the thioesterase (TE)-catalyzed cyclorelease of epothilone from the EpoF protein. It has been unclear whether isolated PKS TE domains could exhibit macrolactonization activity. Here we demonstrate that the excised epothilone TE domain can catalyze the efficient cyclization of the N-acetylcysteamine thioester of seco-epothilone C to generate epothilone C (kcat/KM = 0.41 +/- 0.03 min-1 mM-1). The TE domain also catalyzes the hydrolysis of both the N-acetylcysteamine thioester of seco-epothilone C (kcat = 0.087 +/- 0.005 min-1, KM = 291 +/- 53 muM) and that of the epothilone C (kcat = 0.67 +/- 0.01 min-1, KM = 117 +/- 5 muM) to form seco-epothilone C. PMID- 12643696 TI - Assembly of resorcinarene capsules in wet solvents. AB - Resorcinarenes assemble in wet chloroform or benzene to form hexameric capsules, resembling inflated cubes or volleyballs. NMR methods are used to determine the number of solvent molecules detained inside; eight molecules of benzene are encapsulated. PMID- 12643697 TI - Nanometer distance measurements on RNA using PELDOR. AB - A rapid increase in RNA structural studies has been seen in the past few years, in part due to the interest in the multitude of cellular functions performed by RNA. Electron paramagnetic resonance spectroscopy is a useful technique for studying biopolymer structure under physiologically relevant conditions. We present herein the use of pulsed electron double resonance for the determination of a 35 +/- 2 A distance between two spin label nitroxides that were linked to specific positions within an RNA. PMID- 12643698 TI - A rare earth-DOTA-binding antibody: probe properties and binding affinity across the lanthanide series. AB - An antibody that binds rare earth complexes selectively could be used as a docking station for a set of probe molecules, of particular interest for medical imaging and therapy. The rare earths are rich in probe properties, such as the paramagnetism of Gd, the luminescence of Tb and Eu, and the nuclear properties of Lu and Y. We find that antibody 2D12.5, initially developed to bind analogues of Y-DOTA (1,4,7,10-tetraazacyclododecane-N,N',N' ',N' ''-tetraacetic acid) for radiotherapy, binds not only Y-DOTA analogues but also analogous DOTA complexes of all of the lanthanides. Surprisingly, chelates of some metals such as Gd3+ bind more tightly than the original Y3+ complex. When the shape of the complex is perturbed by either increasing or decreasing the radius of the lanthanide ion, the thermodynamic stability of the protein-ligand complex changes in a regular fashion. The behavior of DeltaDeltaG as a function of ionic radius fits a parabola, as might be expected for a system that behaves in a thermodynamically elastic way. The broad specificity and high affinity of this antibody for all rare earth-DOTA complexes make it particularly interesting for applications that take advantage of the unique characteristics of lanthanides. For example, UV excitation of the Tb-DOTA-2D12.5 complex leads to energy transfer from aromatic side chains of the antibody to bound Tb-DOTA, enhancing green terbium luminescence >104 relative to unbound Tb-DOTA. PMID- 12643699 TI - A new approach in 1D and 2D 13C high-resolution solid-state NMR spectroscopy of paramagnetic organometallic complexes by very fast magic-angle spinning. AB - Novel 1D and multidimensional solid-state NMR (SSNMR) methods using very fast magic-angle spinning (VFMAS) (spinning speed > 20 kHz) for performing 13C high resolution SSNMR of paramagnetic organometallic complexes are discussed. VFMAS removes a majority of 13C-1H and 1H-1H dipolar couplings, which are often difficult to remove by RF pulse techniques in paramagnetic complexes because of large paramagnetic shifts. In the first systematic approach using the unique feature of VFMAS for paramagnetic complexes, we demonstrate a means of obtaining well-resolved 1D and multidimensional 13C SSNMR spectra, sensitivity enhancements via cross polarization, and signal assignments, and applications of dipolar recoupling methods for nonlabeled paramagnetic organometallic complexes of moderate paramagnetic shifts ( approximately 800 ppm). Experimental results for powder samples of small nonlabeled coordination complexes at 1H frequencies of 400.2-400.3 MHz show that highly resolved 13C SSNMR spectra can be obtained under VFMAS, without requirements of 1H decoupling. Sensitivity enhancement in 13C SSNMR via cross polarization from 1H spins was demonstrated with an amplitude sweep high-power CP sequence using strong RF fields ( approximately 100 kHz) available in the VFMAS probe. 13C CPMAS spectra of nonlabeled Cu(II)(dl alanine)2.(H2O) and V(III)(acetylacetonate)3 (V(acac)3) show that it is possible to obtain high-resolution spectra for a small quantity ( approximately 15 mg) of nonlabeled paramagnetic organometal complexes within a few minutes under VFMAS. Experiments on Cu(II)(dl-alanine)2.(H2O) demonstrated that 1H-13C dipolar recoupling for paramagnetic organometal complexes can be performed under VFMAS by application of rotor-synchronous pi-pulses to 1H and 13C spins. The results also showed that signal assignments for 13CH, 13CH3, and 13CO groups in paramagnetic complexes are possible on the basis of the amount of 13C-1H dipolar dephasing induced by dipolar recoupling. Furthermore, the experimental 2D 13C/1H chemical shift correlation NMR spectrum obtained for nonlabeled V(acac)3 exhibits well resolved lines, which overlap in 1D 13C and 1H spectra. Signals for different chemical groups in the 2D spectrum are distinguished by the 13C-1H dipolar dephasing method combined with the 2D 13C/1H correlation NMR. The assignments offer information on the existence of nonequivalent ligands in the coordination complex in solids, without requiring a single-crystal sample. PMID- 12643700 TI - Synthesis of InP nanotubes. AB - Indium phosphide (InP) nanotubes have been synthesized via the vapor-liquid-solid (VLS) growth mechanism. The nanotubes are crystalline and have the (bulk) zinc blende structure and therefore represent a new class of tube materials. The tubes show photoluminescence, which is considerably blue-shifted with respect to bulk emission, indicating that the optical properties are not dominated by defect states. They are formed at higher temperatures than those at which nanowires are fabricated. A simple model for the formation of the nanotubes is presented. The wall thickness can be controlled by the synthesis temperature and is in the range of 2-20 nm. PMID- 12643701 TI - Catalytic asymmetric reductive coupling of alkynes and aldehydes: enantioselective synthesis of allylic alcohols and alpha-hydroxy ketones. AB - A highly enantioselective method for catalytic reductive coupling of alkynes and aldehydes is described. Allylic alcohols are afforded with complete E/Z selectivity, generally >95:5 regioselectivity, and in up to 96% ee. In conjunction with ozonolysis, this process is complementary to existing methods of enantioselective alpha-hydroxy ketone synthesis. PMID- 12643702 TI - Direct preparation of nanoporous carbon by nanocasting. AB - Nanoporous carbon with narrow pore size distribution was prepared via a nanocasting technique by direct carbonization of cyclodextrin-silica organic inorganic hybrid composite. The obtained carbon exhibited very high BET surface area and high pore volume. PMID- 12643703 TI - An outer-sphere two-electron platinum reagent. AB - A strategy for designing cooperative outer-sphere two-electron platinum reagents is demonstrated. The novel platinum(II) complex, [Pt(tpy)(pip2NCN)][BF4] (1(BF4 )) (tpy = 2,2':6',2' '-terpyridine, pip2NCN- = 2,6-(CH2N(CH2)5)2-C6H3-), in which the metal is bonded to two pincer type ligands, has been prepared. Treatment of 1 with protic acid results in protonation of the pendant piperdyl groups, allowing for the isolation of [Pt(tpy)(pip2NCNH2)][PF6]3 (2(PF6-)3). 1H NMR spectra of 1 and 2 establish that in each complex the terpyridyl ligand is tridentate, whereas the piperdyl ligand is monodentate, bonded to platinum through the phenyl ring. The structure of the protonated complex was confirmed by an X-ray crystallographic study of crystals of 2(Cl-)3.4H2O. The cyclic voltammagram of 1 exhibits two reversible one-electron reduction waves at E degrees ' = -0.98 V and E degrees ' = -1.50 V (E degrees ' = (Epc + Epa)/2), with a DeltaEp of 65 and 61 mV, respectively. In contrast to other Pt(II) complexes, including 2, this complex also undergoes a nearly reversible two-electron oxidation process at E degrees ' = 0.40 V (DeltaEp = 43 mV, 0.01 V/s). The accumulated data are consistent with the unusual ligand architecture of 1 being capable of stabilizing and allowing for facile interconversion between the Pt(II) and Pt(IV) oxidation states. PMID- 12643704 TI - Aromatic delta-peptides. AB - Oligoamides of 8-amino-2-carboxy-quinoline adopt a very stable helical conformation characterized in solution by 1H NMR and in the solid state by single crystal X-ray diffraction. The helix comprises only 2.5 units per turn, which represents the highest curvature achieved by aromatic oligoamides until now. Monomers possessing alkoxy substituents in position 4 are easily available from 2 nitroaniline and dimethyl acetylenedicarboxylate. PMID- 12643705 TI - Polymer-directed synthesis of penniform BaWO4 nanostructures in reverse micelles. AB - Novel penniform superstructures of BaWO4 nanowires have been successfully synthesized in catanionic reverse micelles by using a double-hydrophilic block copolymer as the directing agent. This synthetic method is very simple, mild, and controllable, and it provides a novel method for direct solution-growth of hierarchical nanostructures based on inorganic nanowires. PMID- 12643706 TI - Amplified DNA sensing and immunosensing by the rotation of functional magnetic particles. AB - Amplified chemiluminescent detection of DNA-complementary DNA or of antigen antibody interactions is accomplished in the presence of rotating functionalized magnetic particles. PMID- 12643707 TI - Artificial viruses and their application to gene delivery. Size-controlled gene coating with glycocluster nanoparticles. AB - Number- and size-controlled macromolecular associations are common in biology with viruses as a typical example. We report here a novel example of artificial viruses, in which the double-helical DNA is coated with 4-nm sized neutral glycocluster nanoparticles (GNPs) with a coating stoichiometry of approximately 2 GNPs per helical pitch (10 base pairs), where GNP arises from micellization of a cone-shaped, quadruple-chain glycocluster amphiphile having eight saccharide moieties with beta-glucoside termini on the calix[4]resorcarene macrocycle. The resulting "glycoviruses" are compactly packed (54 nm in the case of 7040 base pair plasmid pCMVluc), are well charge-shielded (zeta congruent with approximately 0 mV), and effectively transfect cell cultures without notable cytotoxicity. The use of artificial viral vectors thus allows a new (nonamine/noncationic/nonpolymeric) access to gene delivery, a potential but still tough subject which has been studied extensively over the last 15 years by using viral or amine-based cationic vectors. The remarkable adhesion-manipulation ability of saccharide clusters also provides a strategy of bottom-up construction of nanometric or mesoscopic sizes. PMID- 12643708 TI - NMR structure of two novel polyethylene glycol conjugates of the human growth hormone-releasing factor, hGRF(1-29)-NH2. AB - Two novel mono-PEGylated derivatives of hGRF(1-29)-NH(2) [human growth hormone releasing factor, fragment 1-29] have been synthesized by regio-specific conjugation of Lys(12) or Lys(21) to a monomethoxy-PEG(5000) chain (compounds Lys(12)PEG-GRF and Lys(21)PEG-GRF). The PEG moiety has been covalently linked at the amino group of a norleucine residue via a carbamate bond. The Lys(12)PEG-GRF regioisomer was found to be slightly less active in vitro than both the unmodified peptide and Lys(21)PEG-GRF. To assess whether the differences in the biological activity of the PEGylated analogues could be related to conformational rearrangements induced by the PEG moiety, the structure of these PEGylated derivatives has been worked out (TFE solution) by means of NMR spectroscopy and molecular dynamics. Secondary structure shifts, hydrogen/deuterium exchange kinetics, temperature coefficients of amide protons, and NOE-based molecular models point out that hGRF(1-29)-NH(2), Lys(21)PEG-GRF and Lys(12)PEG-GRF share a remarkably similar pattern of secondary structure. All three compounds adopt an alpha-helix conformation which spans the whole length of the molecule, and which becomes increasingly rigid on going from the N-terminus to the C-terminus. Residues Lys(12) and Lys(21) are enclosed in all the compounds considered into well-defined alpha-helical domains, indicating that PEGylation either at Lys(12) or Lys(21) does not alter the tendency of the peptide to adopt a stable alpha helix conformation, nor does it induce appreciable conformational mobility in the proximity of the PEGylation sites. No significant variation of the amphiphilic organization of the alpha-helix is observed among the three peptides. Therefore, the different biological activities observed for the PEGylated analogues are not due to conformational effects, but are rather due to sterical hindrance effects. The relationship between the biological activitiy of the mono-PEGylated derivatives and sterical hindrance is discussed in terms of the topology of interaction between hGRF(1-29)-NH(2) and its receptor. PMID- 12643710 TI - Design of a new fluorescent cofactor for DNA methyltransferases and sequence specific labeling of DNA. AB - Sequence-specific labeling of DNA is of immense interest for analytical and functional studies of DNA. We present a novel approach for sequence-specific labeling of DNA using a newly designed fluorescent cofactor for the DNA methyltransferase from Thermus aquaticus (M.TaqI). Naturally, M.TaqI catalyzes the nucleophilic attack of the exocyclic amino group of adenine within the double stranded 5'-TCGA-3' DNA sequence onto the methyl group of the cofactor S-adenosyl L-methionine (AdoMet) leading to methyl group transfer. The design of a new fluorescent cofactor for covalent labeling of DNA was based on three criteria: (1) Replacement of the methionine side chain of the natural cofactor AdoMet by an aziridinyl residue leads to M.TaqI-catalyzed nucleophilic ring opening and coupling of the whole nucleoside to DNA. (2) The adenosyl moiety is the molecular anchor for cofactor binding. (3) Attachment of a fluorophore via a flexible linker to the 8-position of the adenosyl moiety does not block cofactor binding. According to these criteria the new fluorescent cofactor 8-amino[1''-(N''-dansyl) 4''-aminobutyl]-5'-(1-aziridinyl)-5'-deoxyadenosine (3) was synthesized. 3 binds about 4-fold better than the natural cofactor AdoMet to M.TaqI and is coupled with a short duplex oligodeoxynucleotide by M.TaqI. The identity of the expected modified nucleoside was verified by electrospray ionization mass spectrometry after enzymatic fragmentation of the product duplex. In addition, the new cofactor 3 was used to sequence-specifically label plasmid DNA in a M.TaqI catalyzed reaction. PMID- 12643709 TI - Highly efficient sequence-specific DNA interstrand cross-linking by pyrrole/imidazole CPI conjugates. AB - We have developed a novel type of DNA interstrand cross-linking agent by synthesizing dimers of a pyrrole (Py)/imidazole (Im)-diamide-CPI conjugate, ImPyLDu86 (1), connected using seven different linkers. The tetramethylene linker compound, 7b, efficiently produces DNA interstrand cross-links at the nine-base pair sequence, 5'-PyGGC(T/A)GCCPu-3', only in the presence of a partner triamide, ImImPy. For efficient cross-linking by 7b with ImImPy, one A.T base pair between two recognition sites was required to accommodate the linker region. Elimination of the A.T base pair and insertion of an additional A.T base pair and substitution with a G.C base pair significantly reduced the degree of cross linking. The sequence specificity of the interstrand cross-linking by 7b was also examined in the presence of various triamides. The presence of ImImIm slightly reduced the formation of a cross-linked product compared to ImImPy. The mismatch partners, ImPyPy and PyImPy, did not produce an interstrand cross-link product with 7b, whereas ImPyPy and PyImPy induced efficient alkylation at their matching site with 7b. The interstrand cross-linking abilities of 7b were further examined using denaturing polyacrylamide gel electrophoresis with 5'-Texas Red-labeled 400 and 67-bp DNA fragments. The sequencing gel analysis of the 400-bp DNA fragment with ImImPy demonstrated that 7b alkylates several sites on the top and bottom strands, including one interstrand cross-linking match site, 5'-PyGGC(T/A)GCCPu 3'. To obtain direct evidence of interstrand cross-linkages on longer DNA fragments, a simple method using biotin-labeled complementary strands was developed, which produced a band corresponding to the interstrand cross-linked site on both top and bottom strands. Densitometric analysis indicated that the contribution of the interstrand cross-link in the observed alkylation bands was approximately 40%. This compound efficiently cross-linked both strands at the target sequence. The present system consisted of a 1:2 complex of the alkylating agent and its partner ImImPy and caused an interstrand cross-linking in a sequence-specific fashion according to the base-pair recognition rule of Py-Im polyamides. PMID- 12643712 TI - Electronic control of the regiochemistry in the Heck reaction. AB - The migratory insertion of propene into the Pd-phenyl bond of a set of neutral palladium(II) complexes with N-N, P-O, and N-O chelating ligands has been investigated by using density functional methods. In contrast to cationic palladium(II)-diimine complexes, which favor 1,2-insertion, the more electron rich neutral complexes lead to 2,1-insertion. Trans influences are substantial in the unsymmetrical ligand systems, primarily affecting the insertion barrier. Introducing steric effects leads toward improving 1,2-selectivity, due to destabilization of the 2,1-transition state. Electronic and steric influences can be systematically combined to control the regiochemistry in the Heck reaction. PMID- 12643711 TI - Anomalous binding profile of phenylboronic acid with N-acetylneuraminic acid (Neu5Ac) in aqueous solution with varying pH. AB - Borates are known to interact with carbohydrate moieties expressed on the surface of biological membranes of a variety of cells, viruses, bacteria, and fungi. This study revealed the anomalous binding profile of borate in aqueous solution with N acetylneuraminic acid (Neu5Ac, sialic acid) as a potential receptor site on the surfaces of biological membranes using (11)B, (1)H, (13)C, and (15)N nuclear magnetic resonance spectroscopies. 3-(Propionamido)phenylboronic acid (PAPBA) was chosen as the model borate compound. The equilibrium constant (K) for Neu5Ac binding to PAPBA was compared with those for glucose, mannose, and galactose, which are the major carbohydrate constituents of glycoproteins and glycolipids expressed on biological membranes. In the Neu5Ac/PAPBA system, the unusual pH dependency of the K values, a decrease in K with increasing pH, was observed, suggesting the formation of a trigonal-formed complex stabilized by the coordination of an amide group of Neu5Ac at the C-5 position to the boron atom, forming intramolecular B-N or B-O bonding. Furthermore, the anomalously high complexing ability at physiological pH 7.4 was confirmed for this system, with the K value 37.6 which is approximately 7 times higher than that for glucose. This exceptionally high value of K at physiological pH, compared to those of other sugars, strongly suggests that the boronic acid selectively recognizes the Neu5Ac residues of the glycosylated components including glycoproteins and gangliosides existing on the surface of the biological membranes. PMID- 12643713 TI - Amine- and ether-chelated aryllithium reagents-structure and dynamics. AB - Chelation and aggregation in phenyllithium reagents with potential 6- and 7-ring chelating amine (2, 3) and 5-, 6-, and 7-ring chelating ether (4, 5, 6) ortho substituents have been examined utilizing variable temperature (6)Li and (13)C NMR spectroscopy, (6)Li and (15)N isotope labeling, and the effects of solvent additives. The 5- and 6-ring ether chelates (4, 5) compete well with THF, but the 6-ring amine chelate (2) barely does, and 7-ring amine chelate (3) does not. Compared to model compounds (e.g., 2-ethylphenyllithium 7), which are largely monomeric in THF, the chelated compounds all show enhanced dimerization (as measured by K = [D]/[M](2)) by factors ranging from 40 (for 6) to more than 200 000 (for 4 and 5). Chelation isomers are seen for the dimers of 5 and 6, but a chelate structure could be assigned only for 2-(2 dimethylaminoethyl)phenyllithium (2), which has an A-type structure (both amino groups chelated to the same lithium in the dimer) based on NMR coupling in the (15)N, (6)Li labeled compound. Unlike the dimer, the monomer of 2 is not detectably chelated. With the exception of 2-(methoxymethyl)phenyllithium (4), which forms an open dimer (12) and a pentacoordinate monomer (13), the lithium reagents all form monomeric nonchelated adducts with PMDTA. PMID- 12643714 TI - Supramolecular pseudorotaxane polymers from complementary pairs of homoditopic molecules. AB - Self-assembly of supramolecular pseudorotaxane polymers from complementary homoditopic building blocks comprised of bis(dibenzo-24-crown-8) esters derived from the hydroxymethyl crown ether and aliphatic diacid chlorides (CxC, x = number of methylene units in the diacid segment) and 1,10-bis[p (benzylammoniomethyl)phenoxy]alkane bis(hexafluorophosphate)s (AyA, y = number of methylene units in the linker) has been studied. (1)H NMR spectroscopic studies of bis[(2-dibenzo-24-crown-8)methyl] sebacate (C8C) with dibenzylammonium hexafluorophosphate (6) showed that the two binding sites of the ditopic host are equivalent and independent (no positive or negative cooperativity). Likewise the binding sites in 1,10-bis[p-(benzylammoniomethyl)phenoxy]decane bis(hexafluorophosphate) (A10A) were shown to behave independently with dibenzo 24-crown-8 (1a). Then using (1)H NMR spectroscopy on dilute equimolar solutions (<1 mM) of CxC and AyA association constants were estimated for the formation of the linear (lin-CxC*AyA) and cyclic (cyc-CxC*AyA) dimers, thus enabling effective molarities to be estimated for the various systems. Finally (1)H NMR spectroscopy was used to semiquantitatively or qualitatively demonstrate the formation of linear supramolecular polymers lin-[CxC*AyA](n) in more concentrated solutions (up to 2.0 M) of the complementary pairs of CxC and AyA. The sizes of the assemblies (n values) are not as great as the dilute solution studies predict; this is attributed to the deleterious effect of ionic strength and exo complexation at high concentrations. However, as expected from the dilute solution results, linear extension is indeed favored with the longer building blocks, meaning that "monomer" end-to-end distance is a key factor in reducing the amount of cyclic species that form. Viscosity experiments clearly demonstrate the formation of large noncovalent polymers lin-[CxC*AyA](n) in concentrated solutions. Cohesive film and fiber formation also indicate that supramolecular polymers of sufficient size to enable entanglement self-assemble in these solutions. PMID- 12643715 TI - Application of chiral mixed phosphorus/sulfur ligands to enantioselective rhodium catalyzed dehydroamino acid hydrogenation and ketone hydrosilylation processes. AB - Chiral mixed phosphorus/sulfur ligands 1-3 have been shown to be effective in enantioselective Rh-catalyzed dehydroamino acid hydrogenation and ketone hydrosilylation reactions (eqs 1, 2). After assaying the influence of the substituents at sulfur, the substituents on the ligand backbone, the relative stereochemistry within the ligand backbone, and the substituents at phosphorus, ligands 2c (R = 3,5-dimethylphenyl) and 3 were found to be optimal in the Rh catalyzed hydrogenation of a variety of alpha-acylaminoacrylates in high enantioselectivity (89-97% ee). A similar optimization of the catalyst for the Rh catalyzed hydrosilylation of ketones showed that ligand 3 afforded the highest enantioselectivities for a wide variety of aryl alkyl and dialkyl ketones (up to 99% ee). A model for asymmetric induction in the hydrogenation reaction is discussed in the context of existing models, based on the absolute stereochemistry of the products and the X-ray crystal structures of catalyst precursors and intermediates. PMID- 12643716 TI - Modeling H-bonding and solvent effects in the alkylation of pyrimidine bases by a prototype quinone methide: a DFT study. AB - Nucleophilicity of NH(2), N3, and O(2) centers of cytosine toward a model quinone methide (o-QM) as alkylating agent has been studied using DFT computational analysis [at the B3LYP/6-311+G(d,p) level]. Specific and bulk effects of water (by C-PCM model) on the alkylation pathways have been evaluated by analyzing both unassisted and water-assisted reaction mechanisms. An ancillary water molecule, H bonded to the alkylating agent, may interact monofunctionally with the o-QM oxygen atom (passive mechanisms) or may participate bifunctionally in cyclic hydrogen-bonded structures as a proton shuttle (active mechanisms). A comparison of the unassisted with the water-assisted reaction mechanisms has been made on the basis of activation Gibbs free energies (DeltaG(++)). The gas-phase alkylation reaction at N3 does proceed through a passive mechanism that is preferred over both the active (by -6.3 kcal mol(-1)) and the unassisted process. In contrast, in the gas phase, the active assisted processes at NH(2) and O(2) centers are both favored over their unassisted counterparts by -4.0 and -2.2 kcal mol(-1), respectively. The catalytic effect of a water molecule, in gas phase, reduces the gap between the TSs of the O(2) and NH(2) reaction pathways, but the former remains more stable. Water bulk effect significantly modifies the relative importance of the unassisted and water-assisted alkylation mechanisms, favoring the former, in comparison to the gas-phase reactions. In particular, the unassisted alkylation becomes the preferred mechanism for the reaction at both the exocyclic (NH(2)) and the heterocyclic (N3) nitrogen atoms. By contrast, alkylation at the cytosine oxygen atom is a water-catalyzed process, since in water the active water-assisted mechanism is still favored. As far as competition, among all the possible mechanisms, our calculations unambiguously suggest that the most nucleophilic site both in gas phase (naked reagents: N3 >> O(2) >or= NH(2)) and in water solution (solvated reagents: N3 >> NH(2) >> O(2)) is the heterocyclic nitrogen atom (N3) (DeltaG(++)(gas) = +7.1 kcal mol(-1), and DeltaG(++)(solv) = +13.7 kcal mol(-1)). Our investigation explains the high reactivity and selectivity of the cytosine moiety toward o-QM-like structures both in deoxymononucleoside and in a single-stranded DNA, on the basis of strong H-bonding interactions between reactants and solvent bulk effect. It also offers two general reactivity models in water, uncatalyzed and active water-catalyzed mechanisms (for nitrogen and oxygen nucleophiles, respectively), which should provide a general tool for the planning of nucleic acid modification. PMID- 12643717 TI - Ladderlike ferromagnetic spin coupling network on a pi-conjugated pendant polyradical. AB - A poly(9,10-anthryleneethynylene)-based polyradical with two pendant stable phenoxyls in one anthracene skeleton was newly synthesized via polymerization of the corresponding bromoethynylanthracene monomer using a Pd(0) catalyst. The average molecular weight of the polymer reached M(n) = 5 x 10(3) and was soluble in common organic solvents. The polyradical was prepared from the corresponding hydroxyl precursor polymer and was appropriately stable at room temperature. The ESR spectrum of the corresponding monomeric radical suggested an effectively delocalized spin density distribution on the backbone anthracene. The magnetization and the static magnetic susceptibility of the polyradical were measured using a SQUID magnetometer. The large average spin quantum number (S = (5)/(2)) of the polyradical indicated that the ferromagnetic spin coupling network of the polyradical had spread throughout the pi-conjugated chain and that it was considerably insensitive to spin defects. PMID- 12643718 TI - Photodissociation of the phosphine-substituted transition metal carbonyl complexes Cr(CO)(5)L and Fe(CO)(4)L: a theoretical study. AB - The photochemistry of the phosphine-substituted transition metal carbonyl complexes Cr(CO)(5)PH(3) and ax-Fe(CO)(4)PH(3) is studied with time-dependent DFT theory to explore the propensity of the excited molecules to expel their ligands. The influence of the PH(3) ligand on the properties of these complexes is compared with the photodissociation behavior of the binary carbonyl complexes Cr(CO)(6) and Fe(CO)(5). The lowest excited states of Cr(CO)(5)PH(3) are metal-to ligand charge transfer (MLCT) states, of which the first three are repulsive for PH(3) but modestly bonding for the axial and equatorial CO ligands. The repulsive nature is due to mixing of the initial MLCT state with a ligand field (LF) state. A barrier is encountered along the dissociation coordinate if the avoided crossing between these states occurs beyond the equilibrium distance. This is the case for expulsion of CO but not for the PH(3) group as the avoided state crossing occurs within the equilibrium Cr-P distance. The lowest excited state of ax-Fe(CO)(4)PH(3) is a LF state that is repulsive for both PH(3) and the axial CO. Excited-state quantum dynamics calculations for this state show a branching ratio of 99 to 1 for expulsion of the axial phosphine ligand over an axial CO ligand. The nature of the phosphorus ligand in these Cr and Fe complexes is only of modest importance. Complexes containing the three-membered phosphirane or unsaturated phosphirene rings have dissociation curves for their lowest excited states that are similar to those having a PH(3) ligand. Analysis of their ground state Cr-P bond properties in conjunction with frontier orbital arguments indicate these small heterocyclic groups to differ from the PH(3) group mainly by their enhanced sigma-donating ability. All calculations indicate that the excited Cr(CO)(5)L and Fe(CO)(4)L molecules (L = PH(3), PC(2)H(5), and PC(2)H(3)) prefer dissociation of their phosphorus substituent over that of an CO ligand. This suggests that the photochemical approach may be a viable complement to the ligand exchange and redox methods that are currently employed to demetalate transition metal complexed organophosphorus compounds. PMID- 12643719 TI - Redoxable nanosheet crystallites of MnO2 derived via delamination of a layered manganese oxide. AB - This paper reports on the swelling and exfoliation behavior of a layered protonic manganese oxide, H(0.13)MnO(2).0.7H(2)O, in a solution of tetrabutylammonium (TBA) hydroxide and the formation and characterizations of unilamellar two dimensional crystallites of MnO(2). At low doses of TBA ions, layered manganese oxide was observed to undergo normal intercalation, yielding a TBA intercalated phase with a gallery height of 1.25 nm. With a large excess of TBA ions, osmotic swelling occurred, giving rise to a very large intersheet separation of 3.5-7 nm. In an intermediate TBA concentration range, the sample exhibited a broad X-ray diffraction profile with superimposed diffraction features due to intercalation and osmotic swelling. The component responsible for the broad profile was isolated by centrifuging the mixture twice at different speeds, and the recovered colloid was identified as a pile of MnO(2) nanosheets, corresponding to the individual host layer of the precursor layered manganese oxide. Observations by transmission electron microscopy and atomic force microscopy revealed high two dimensional anisotropy with a lateral dimension of submicrometers and a thickness of approximately 0.8 nm. The nanosheet exhibited broad optical absorption with a peak at 374 nm (epsilon = 1.13 x 10(4) mol(-1) dm(3) cm(-1)). The restacking process of the colloidal MnO(2) nanosheets was followed by aging the colloid at a relative humidity of 95%. The broad diffraction pattern due to the exfoliated sheets weakened with time and eventually resolved into two sharp distinct profiles attributable to a TBA intercalation compound with an intersheet spacing of 1.72 nm and an osmotically swollen hydrate with >10 nm at a very early stage. As drying progressed, the former phase became more abundant without a change in interlayer distance, while the degree of swelling of the latter phase gradually decreased to 2.7 nm that remained unchanged on further aging. Subsequent drying at a lower humidity collapsed the 2.7 nm phase. The resulting single 1.72 nm phase was dehydrated by heating at 150 degrees C to produce a phase with a contracted interlayer spacing of 1.3 nm. PMID- 12643720 TI - Single-molecule magnets: two-electron reduced version of a Mn12 complex and environmental influences on the magnetization relaxation of (PPh4)(2)[Mn(12)O(12)(O(2)CCHCl2)(16)(H2O)4]. AB - The complex [Mn(12)O(12)(O(2)CCHCl(2))(16)(H(2)O)(4)] (2) in MeCN exhibits three quasi-reversible one-electron reduction processes at significantly higher potentials than [Mn(12)O(12)(O(2)CMe)(16)(H(2)O)(4)] (1). This has allowed the two-electron reduced version of 2 to be generated and isolated. Reaction of 2 with one and two equivalents of PPh(4)I led to isolation of (PPh(4))[Mn(12)O(12)(O(2)CCHCl(2))(16)(H(2)O)(4)] (3) and (PPh(4))(2)[Mn(12)O(12)(O(2)CCHCl(2))(16)(H(2)O)(4)] (4), respectively. The latter represents a new isolated oxidation level of the Mn(12) family of single molecule magnets (SMMs). Crystallization from CH(2)Cl(2)/hexanes yields a mixture of two crystal forms, 4.4CH(2)Cl(2).H(2)O (4a) and 4.6CH(2)Cl(2) (4b), both of which have been structurally characterized as triclinic and monoclinic, respectively. The molecular structures are very similar, with the added electrons localized on former Mn(III) ions to give a trapped-valence 2Mn(II), 6Mn(III), 4Mn(IV) oxidation state description. Dried solid analyzed as unsolvated 4. (1)H NMR spectral data in CD(2)Cl(2) confirm that 4 retains its solid-state structure in solution. Bulk DC magnetization data for dried 4 in the 1.80-4.00 K and 10-70 kG ranges were fit to give S = 10, D = -0.275 cm(-1), g = 2.00 and |D|/g = 0.14 cm(-1), where D is the axial zero-field splitting (anisotropy) parameter. Complexes 4a and 4b give resolvable frequency-dependent out-of-phase (chi(M)'') signals in AC susceptibility studies resulting from the magnetization relaxation of SMMs. Relaxation rate vs T data to 1.8 K obtained from the chi(M)'' vs temperature studies were supplemented with rate vs T data measured to lower temperatures via magnetization vs time decay data, and these were fit to the Arrhenius equation to give the effective barrier to relaxation (U(eff)). The U(eff) values are 18.5 and 30.3 K for 4a and 4b, respectively. A similar analysis for dried 4 using AC data gave U(eff) = 32 K. Magnetization vs DC field sweeps on single crystals of 4a and 4b gave hysteresis loops containing steps due to quantum tunneling of magnetization (QTM). The step separations yielded |D|/g values of 0.087 and 0.14 cm(-1) for 4a and 4b, respectively, suggesting that the differences in U(eff) are primarily caused by changes to D. The combined work demonstrates the sensitivity of the magnetic properties of these new [Mn(12)](2-) SMMs to subtle differences in their environment as determined by the precise packing, solvent molecules, and overall crystal symmetry (space group) and represents an important caveat to workers in the field. PMID- 12643721 TI - Suppression of binary nucleation in amorphous La-Fe-Sb mixtures. AB - The nucleation energy of a series of La(x)Fe(y)Sb(z) modulated elemental reactants was measured as a function of the Fe/Sb ratio over a large composition range while holding the La content constant. The nucleation energy of the ternary compound La(0.5)Fe(4)Sb(12) with the skutterudite crystal structure was found to depend very strongly on the Fe/Sb ratio in the modulated elemental reactant, with a higher nucleation energy as the Fe/Sb ratio is moved away from the 1:3 stoichiometric value. When the results of this study are compared with those from Fe(y)Sb(z) modulated reactants, the addition of lanthanum was found to suppress the nucleation of FeSb(2), thereby broadening the Fe/Sb composition range in which the ternary skutterudite compound La(x)Fe(4)Sb(12) nucleates. This suppression of nucleation of a binary phase on addition of a ternary component to an amorphous intermediate is in agreement with theoretical arguments. The observed suppression of nucleation also provides rational for the observed nucleation of metastable ternary and higher-order compounds from homogeneous amorphous reactants. PMID- 12643722 TI - Understanding the formation of new clusters of alkali and alkaline Earth metals: a new synthetic approach, single-crystal structures, and theoretical calculations. AB - A new synthetic approach, reacting alkaline earth metal iodides with butyllithium, lithium hydroxide, and/or lithium butoxide under salt elimination, is presented, giving access to some interesting clusters of calcium, strontium, and barium, partially in combination with lithium. The so far largest calcium cluster Li[[Ca(7)(mu(3)-OH)(8)I(6)(thf)(12)](2)(mu(2)-I)].3THF, 4, and the new strontium compound [Sr(3)I(3)(OH)(2)(thf)(9)]I, 5, are shown to feature common building blocks of OH-capped M(3) triangles. On the basis of mainly electrostatic interactions, these clusters are not volatile. By introducing LiO(t)Bu, the two clusters [IM(O(t)Bu)(4)[Li(thf)](4)(OH)] (6, M = Sr; 7, M = Ba) are prepared, 7 exhibiting volatility as an important physical property, which makes it a potential precursor for chemical vapor deposition. The structural relationship between 4, 5, 6, and 7 and their respective starting materials is shown, and possible reaction mechanisms are proposed. Exhibiting surprising and new structural motifs, the bonding modes of these clusters are investigated by the electron localization function as well as by ab initio calculations. PMID- 12643723 TI - Mechanism of ketone and alcohol formations from alkenes and alkynes on the head to-head 2-pyridonato-bridged cis-diammineplatinum(III) dinuclear complex. AB - Reactions of the head-to-head 2-pyridonato-bridged cis-diammineplatinum(III) dinuclear complex having nonequivalent two platinum atoms, Pt(N(2)O(2)) and Pt(N(4)), with p-styrenesulfonate, 2-methyl-2-propene-1-sulfonate, 4-penten-1-ol, and 4-pentyn-1-ol were studied kinetically. Under the pseudo first-order reaction conditions that the concentration of the Pt(III) dinuclear complex is much smaller than that of olefin, a consecutive basically four-step reaction was observed: the olefin pi-coordinates preferentially to the Pt(N(2)O(2)) in the first step (step 1), followed by the second pi-coordination of another olefin molecule to the Pt(N(4)) (step 2). In the next step (step 3), the nucleophilic attack of water to the coordinated olefin triggers the pi-sigma bond conversion on the Pt(N(2)O(2)), and the second pi-bonding olefin molecule on the Pt(N(4)) is released. Finally, reductive elimination occurs to the alkyl group on the Pt(N(2)O(2)) to produce the alkyl compound (step 4). The first water substitution with olefin (step 1) occurs to the diaqua and aquahydroxo forms of the complex, whereas the second substitution (step 2) proceeds either on the coordinated OH(-) on the Pt(N(4)) (path a) or on the coordinatively unsaturated five-coordinate intermediate of the Pt(N(4)) (path b), in addition to the common substitution of H(2)O (path c). The reactions of p-styrenesulfonate and 2-methyl-2-propene-1 sulfonate proceed through paths b and c, whereas the reactions of 4-penten-1-ol and 4-pentyn-1-ol proceed through paths a and c. This difference reflects the difference of the trans effect and/or trans influence of the pi-coordinated olefins on the Pt(N(2)O(2)). The pentacoordinate state in path b is employed only by the sulfo-olefins, because these exert stronger trans effect. The steps 3 and 4 reflect the effect of the axial alkyl ligand (R) on the charge localization (R Pt(IV)(N(2)O(2))-Pt(II)(N(4))) and delocalization (R-Pt(III)(N(2)O(2)) Pt(III)(N(4))-OH(2)); when R is p-styrenesulfonate having an electron withdrawing group, the charge localization in the dimer is less pronounced and the water molecule on the Pt(N(4)) atom is retained (R-Pt(III)(N(2)O(2))-Pt(III)(N(4)) OH(2)) in the intermediate state. In both routes, the alkyl group undergoes nucleophilic attack of water, and the oxidized products are released via reductive elimination. PMID- 12643724 TI - Covalent sidewall functionalization of single wall carbon nanotubes. AB - Alkyllithium reagents may be used to attach alkyl groups to the sidewalls of fluoro nanotubes. Thermal gravimetric analysis combined with UV-vis-Nir spectroscopy has been used to provide a quantitative measure of the degree of functionalization. SWNTs prepared using the HiPco process exhibit a higher degree of alkylation than SWNTs from the laser-oven method, indicating that the smaller diameter fluoro tubes are alkylated more readily. The spectral signature of the pristine SWNTs can be regenerated when the alkylated SWNTs are heated in Ar at 500 degrees C, demonstrating that dealkylation occurs at this temperature. TGA-MS analysis using a sample of n-butylated h-SWNTs showed that 1-butene and n-butane are formed during thermolysis. PMID- 12643725 TI - Complexity of coordinative bonding in thallium(I) anthranilates and salicylates. AB - An inventory of the structural chemistry of thallium(I) shows many unexpected, almost random coordination numbers and coordination geometries that appear erratic and inconsistent. This nonstandard behavior is often ascribed to the specific lone-pair characteristics originating from relativistic effects. To provide data on a set of closely related compounds from which simple general rules of coordinative bonding at Tl(+) can be established, three thallium(I) anthranilates and three thallium(I) salicylates have been prepared from Tl(2)CO(3) and the corresponding 2-amino- and 2-hydroxy-benzoic acids and crystallized from aqueous solutions. All six compounds, the simple anthranilate (1) and salicylate (4) and the 3- and 4-methyl-substituted homologues (2, 3 and 5, 6) show different structures with large variations in the coordination motif. The coordination by oxygen in a geometry which covers less than a coordination hemisphere is the only common feature, complemented (only in 1) by a nitrogen coordination and by eta(6)-coordination of one (in 1, 2, 3, 6) or two phenyl rings (in 4). Tl-Tl contacts for which "thallophilic" bonding between closed shell metal atoms could be invoked, are generally very long (close to 4.0 A) or even well beyond the limit of standard van der Waals contacts. Hydrogen bonding is only obvious for the internal contacts of the amino- or hydroxy-benzoate ligands and does not contribute significantly to the assembly of the supramolecular structure which is dominated by oxygen bridges between thallium atoms. With the exception of 5, the formula units Tl[O(2)C(2-R)(3-R')(4 R'')C(6)H(2)] are generally aggregated into dimers of various configurations depending on the relative orientation of the edge-sharing four-membered rings, and these dimers are further linked into strings or columns establishing N-Tl or Tl-O contacts and arene coordination. The drastic changes induced in the structures upon only small variations such as methyl substitution in 3- or 4 position of the ligand suggest that thallium(I) coordination is generally restricted to one hemisphere of nearest neighbors, but is extremely flexible in this realm. The open hemisphere may be partially capped by arene coordination (which is weak at a distance of ca. 3.1 A to the centroid of the ring) or feature very weak thallophilic contacts. PMID- 12643726 TI - The influence of short strong hydrogen bonding on the structure and the physicochemical properties of alkyl-N-iminodiacetic acids in solid state and aqueous systems. AB - Alkyl-N-iminodiacetic acids with varying alkyl chain lengths have been prepared and characterized with respect to structure, acidic properties, and ability to form aggregates in water. The alkyl-N-iminodiacetic acids are the group of ligands with the lowest molecule weight which can be characterized as chelating surfactants, compounds with surface chemical properties which at the same time have a high ability to bind metal ions. The solid alkyl-N-iminodiacetic acids have a unique structure with neutral zwitterionic units linked together to polymer chains through a short strong hydrogen bond, d(O(-H)...O) approximately 2.46 A, and where the nu(O-H) stretching vibration at ca. 720 cm(-1) supports the presence of such a hydrogen bond. The polymer chains are cross-linked together to bilayers through relatively strong hydrogen bonds between ammonium and carboxylate groups, and where the parallel alkyl groups are interdigitating each other; the bilayer surface consists of hydrophilic iminodiacetic acid groups. The acidic properties of monomeric alkyl-N-iminodiacetic acids in water are in the expected ranges with pK(a) values of about 1.7, 2.3, and 10.3. n-Octadecyl-N iminodiacetic acid, present as aggregates in water, displays very acidic properties of the first proton, and a substantially weakened acidity of the second proton, pK(a2) = 5.5-7.5, depending on ionic strength, and pK(a3) = 9.5 10.5. This pattern of the acidic constants strongly indicates that the polymer structure with short strong hydrogen bonds is maintained in the aggregates and that such bonds can exist in aqueous systems if they are supported by a strong and rigid backbone structure, as the bilayers of well-organized long interdigitating alkyl chains in the studied systems. Hydrogenbis(methyl-N iminodiacetic acid) perchlorate precipitates from perchloric acidic solutions of methyl-N-iminodiacetic acid. The structure is built up of dimers of zwitterionic methyl-N-iminodiacetic acid units linked together by an extra proton in a short strong hydrogen bond, d(O(-H)...O) approximately 2.456(6) A, and nu(O-H) = 789 cm(-1). PMID- 12643727 TI - Modeling the reaction mechanisms of the imide formation in an N-(o carboxybenzoyl)-L-amino acid. AB - Reaction mechanisms of the imide formation in an N-(o-carboxybenzoyl)-l-amino acid have been studied using density functional theory. Our results suggest that the reaction route initiated by protonation at the oxygen of the carboxyl group of the amino acid is favored, while those initiated by deprotonation at the oxygen of the carboxyl group of phthalic acid and at the amidic nitrogen are minor pathways. During the dehydration process, water functions as a catalyst. These conclusions are in good agreement with the experimental facts that at highly acidic conditions (hydrogen ion concentration H(0) < -1), imide formation is the most favorable pathway, whereas in the pH range 0-5, cyclization to the imide is not the dominant reaction. Our calculations also show that the carboxyl group of the amino acid is involved in the catalytic reaction in both the favored and minor pathways and that solvent effects have little influence on the reaction barriers. PMID- 12643728 TI - DFT analysis of NMR scalar interactions across the glycosidic bond in DNA. AB - The relationship between the glycosidic torsion angle chi, the three-bond couplings (3)J(C2/4-H1') and (3)J(C6/8-H1'), and the one-bond coupling (1)J(C1' H1') in deoxyribonucleosides and a number of uracil cyclo-nucleosides has been analyzed using density functional theory. The influence of the sugar pucker and the hydroxymethyl conformation has also been considered. The parameters of the Karplus relationships between the three-bond couplings and chi depend strongly on the aromatic base. (3)J(C2/4-H1') reveals different behavior for deoxyadenosine, deoxyguanosine, and deoxycytidine as compared to deoxythymidine and deoxyuridine. In the case of (3)J(C6/8-H1'), an opposite trans to cis ratio of couplings is obtained for pyrimidine nucleosides in contrast to purine nucleosides. The extremes of the Karplus curves are shifted by ca. 10 degrees with respect to syn and anti-periplanar orientations of the coupled nuclei. The change in the sugar pucker from S to N decreases (3)J(C2/4-H1') and (3)J(C6/8-H1'), while increasing (1)J(C1'-H1') for the syn rotamers, whereas all of the trends are reversed for the anti rotamers. The influence of the sugar pucker on (1)J(C1'-H1') is interpreted in terms of interactions between the n(O4'), sigma*(C1'-H1') orbitals. The (1)J(C1'-H1') are related to chi through a generalized Karplus relationship, which combines cos(chi) and cos(2)(chi) functions with mutually different phase shifts that implicitly accounts for a significant portion of the related sugar pucker effects. Most of theoretical (3)J(C2/4-H1') and (3)J(C6/8 H1') for uracil cyclo-nucleosides compare well with available experimental data. (3)J(C6/8-H1') couplings for all C2-bridged nucleosides are up to 3 Hz smaller than in the genuine nucleosides with the corresponding chi, revealing a nonlocal aspect of the spin-spin interactions across the glycosidic bond. Theoretical (1)J(C1'-H1') are underestimated with respect to the experiment by ca. 10% but reproduce the trends in (1)J(C1'-H1') vs chi. PMID- 12643729 TI - Infrared spectroscopy of niobium oxide cluster cations in a molecular beam: identifying the cluster structures. AB - Infrared spectra of niobium oxide cluster cations are measured via IR multiple photon dissociation spectroscopy in the 400-1650 cm(-1) region. The cluster cations are obtained directly from a laser vaporization source and irradiated with the infrared light emitted by a free electron laser. For those oxide clusters that fragment after excitation, the IR spectra are recorded by measuring the cluster intensity changes as a function of the IR wavelength. The spectra of all examined oxide clusters exhibit two main absorption features that can be assigned to vibrations of terminal (Nb=O) or bridging (Nb-O-Nb) oxide groups. For selected clusters DFT calculations at the B3LYP/LACVP* level have been performed and the calculated vibrational spectra are compared to the experimental data to identify the gas phase structures of the clusters. PMID- 12643734 TI - Comment on "in vitro and in vivo evaluation of a Technetium-99m-labeled cyclic RGD peptide as a specific marker of alphaVbeta3 integrin for tumor imaging". PMID- 12643736 TI - An efficient and versatile synthesis of bisPNA-peptide conjugates based on chemoselective oxime formation. AB - Oligomers with two identical peptide nucleic acid sequences joined by a flexible hairpin linker (bisPNA) can stably bind to specific DNA sequences without altering plasmid supercoiling, thus offering a unique opportunity to attach various functional entities to high molecular weight DNA. Current synthetic approaches, however, severely limit the possibility to link peptides or other chemical moieties (i.e., sugars, oligonucleotides, etc.) to bisPNA. Here we report a novel strategy for the synthesis of bisPNA-peptide conjugates in which chemoselective ligation of bisPNA to peptides was accomplished through oxime formation between an oxy-amine-containing peptide and a bisPNA-methyl ketone (complementary modifications can also be used). The described synthesis is highly efficient, does not require a protection strategy, and is carried out under mild aqueous conditions. Through this methodology long peptide sequences in either C to N or N to C polarity can be linked to bisPNA. In addition, this protocol makes the conjugation of cysteine-containing peptides feasible and allows disulfide bond formation to be controlled. This same approach can be exploited to link oligonucleotides, sugars, or other chemical entities to bisPNA. PMID- 12643737 TI - Can nuclear localization signals enhance nuclear localization of plasmid DNA? AB - Nonviral vectors are safer and more cost-effective than viral vectors but are significantly less efficient, and thus, increasing the efficiency of nonviral vectors remains an important objective. One way to overcome this problem is by stimulating the nuclear localization of exogenous genes. Nuclear localization signals (NLSs) are known to be involved in the active transport of exogenous proteins and probes into the nucleus. However, stimulation of nuclear localization of plasmid DNA has yet to be confirmed completely. In the present study, we prepared plasmid DNA-NLS peptide conjugates and adjusted spacer length and number introduced in an attempt to increase transfection efficiency. In comparison to conjugates with unmodified plasmid DNA and short spacers, we found that NLS-plasmid DNA conjugates with covalent bonding by diazo coupling through PEG chain (MW 3400) stimulated complexation with the nuclear transport proteins importin alpha and importin beta. Evaluation of transfection showed higher expression efficiency with plasmid DNA-NLS peptide conjugates than with unmodified plasmids. However, evaluation of intracellular trafficking after microinjection into the cytoplasm showed plasmid DNA-NLS peptide conjugates only within the cytoplasm; there was no NLS-plasmid stimulation of nuclear localization. Our findings suggest that stimulation of plasmid nuclear localization cannot be achieved merely by changing spacer length or chemically modifying plasmid DNA-NLS peptide conjugates. An additional mechanism must be involved. PMID- 12643738 TI - Synthesis of structurally identical fluorine-18 and iodine isotope labeling compounds for comparative imaging. AB - The synthesis of a benzophenone-based labeling compound designed for comparative imaging studies with both in vivo positron emission tomograph (PET) and single photon computed tomography (SPECT) and ex vivo autoradiography is described. The new compound can be labeled with either F-18 or iodine radioisotopes to give two different radioisotopmers: N-[2-fluoro-5-(3-[I-131]iodobenzoyl)benzyl]-2 bromoacetamide (1) and N-[2-[F-18]fluoro-5-(3-iodobenzoyl)benzyl]-2 bromoacetamide (2). Compound 1 and 2 have a 2-bromoacetyl group, which can be used to conjugate with biomolecules through a nucleophilic substitution reaction. Compound 1 was synthesized from the corresponding tributyltin derivatives via an oxidative destannylation reaction, and compound 2 was prepared via a four-step radiosynthesis (nucleophilic aromatic substitution, reduction, oxidation, and alkylation) starting from 4-(N,N,N-trimethylammonio)-3-cyano-3'-iodobenzophenone triflate. A remarkably high radiochemical yield (>90%) was achieved for the F-18 nucleophilic aromatic substitution under mild conditions (room temperature in less than 10 min), indicating the structural advantage of the designed molecule to facilitate the F-18 for trimethylammonium substitution in the presence of two electron-withdrawing groups (nitrile and carbonyl). The overall radiosynthesis time for compound 2 is less than 3 h after end of bombardment (EOB) with an unoptimized radiochemical yield of about 2% (not decay corrected) and specific activity of 0.8 Ci/micromol at EOB. The radiolabeling precursors for compound 1 and 2 were synthesized via a carbon-carbon bond-forming reaction between 2 substituted-5-lithiobenzonitrile and 3-substituted benzaldehyde derivatives. Compounds 1 and 2 should allow us to label biomolecules with F-18 or iodine isotopes and gives structurally identical products, which are expected to have identical biological properties and should be useful for comparative imaging studies. PMID- 12643739 TI - Conjugation of an antisense oligodeoxynucleotide to ribonuclease h results in sequence-specific cleavage and intracellular inhibition of HCV gene expression. AB - A recombinant E. coli ribonuclease H (RNase H) was chemically coupled to an antisense oligodeoxynucleotide (ODN) against the 5'-noncoding region (5'-NCR) of the hepatitis C virus. Purity of the conjugates was confirmed by sodium deodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as a band corresponding to approximately 23 kDa. Conjugate function was tested by the cleavage of a HCV RNA transcript including the 5'-NCR and core region and showed HCV sequence-specific cleavage by the appearance of an expected approximately 1000 nt fragment of RNA. Cleavage was not seen by RNase H alone, or ODN alone. Delivery studies using (32)P- and (125)I-labeling showed that while RNAse H failed to enter cells, the conjugate was efficiently taken into the cells. To assess intracellular effects, a cell line, Huh-7/CMV-NCRCDeltaluc, which expresses HCV mRNA (nt 1-585) fused to a marker gene, was transfected with the conjugate. Reporter gene expression was suppressed by 51.2% with the conjugate compared to only 39.7% by ODN alone, 35.8% by a mixture of RNase H plus ODN, and not at all by RNase H alone. In conclusion, the RNase H-ODN conjugate effectively cleaved an HCV transcript in vitro and inhibited expression of an HCV-marker fusion construct in a liver-derived cell line. PMID- 12643740 TI - Synthesis and biological evaluation of paclitaxel-C225 conjugate as a model for targeted drug delivery. AB - Tumor-targeted drug delivery is an attractive strategy in cancer treatment. We have previously reported a paclitaxel model conjugate using a bombesin receptor recognizing peptide in which the drug cytotoxicity against H1299 human nonsmall cell lung cancer was enhanced compared to unconjugated taxol. In an effort to expand the development of tumor-recognizing taxanes, paclitaxel (PTX, taxol) was conjugated to the anti-epidermal growth factor receptor (anti-EGFR) monoclonal antibody (MAb) Erbitux (C225) to serve as a model MAb-mediated drug delivery compound. Thus, paclitaxel was derivatized at its 2'-hydroxy function by introduction of a succinate linker, and the carboxyl group of the latter was covalently attached to C225 through amide bond formation. The final product conjugate (PTXC225) was analyzed mass spectrometrically for assessment of the drug-to-antibody ratios. Cytotoxicity screening of the drug-antibody conjugate against A431, UM-SCC-1, and UM-SCC-6 cells indicated an enhancement in cytocidal effect of paclitaxel as compared to those of the free drug, the intact antibody, and a physical mixture of the two (the controls). In A431 cells, the conjugate showed 25.2% +/- 2.2% of apoptosis induction as compared to little or no apoptosis caused by the controls. Biodistribution analysis of the PTXC225 in tumor-implanted nude mice and a tyrosine-kinase assay showed that conjugation of the drug did not interfere with the immunoreactivity of the antibody. The 24-h tumor uptake of C225 and PTXC225 were 11.7% +/- 6.0% and 7.1% +/- 3.6% of the injected dose per gram of tissue (%ID/g), respectively, which were not significantly different. Also, in A431-implanted nude mice, the conjugate and C225 showed tumor growth inhibition effects of 57.2% and 41.2%, respectively, against a saline-treated control, which were not significantly different from each other. This lack of difference in the in vivo antitumor activity of the MAb delivered drug and free PTX may be due to either a relatively low dose of the antibody-delivered drug (346 microg/kg), or an untimely release of it, or both. The tumor growth inhibition pattern of the conjugate, however, was identical to that of C225, indicating that the attachment of PTX did not affect the antigen binding and growth inhibitory features of the MAb. These preliminary results demonstrate the potential of tumor-targeted delivery of taxol as a promising strategy in cancer treatment and warrant further work to develop more suitable drug-MAb linkers as well as improved dosage and treatment protocols. PMID- 12643741 TI - Nanoparticulate DNA packaging using terpolymers of poly(lysine-g-(lactide-b ethylene glycol)). AB - Terpolymers of poly(lysine-g-(lactide-b-ethylene glycol)) (pK-pLL-pEG) were synthesized by using ring-opening polymerization and functional end-group grafting. Synthesis was characterized with gel permeation chromatography, proton nuclear magnetic resonance spectroscopy, and a trinitrobenzene sulfonic acid binding assay. Polymer association behavior with DNA was investigated using an ethidium bromide exclusion assay, static light scattering, and scanning electron microscopy. Polylactide molecular weight was varied to investigate its impact on DNA association and resulting complex characteristics. Polylysine ( = 8800, DP = 42) modified with either 7400 or 10 870 pLL-pEG reduced the minimum amount of primary amines necessary for complete condensation by 23% and 48%, respectively, compared to unmodified polylysine (pK42). Complexes formed with the highest molecular weight terpolymer demonstrated significantly (p < 0.1) greater resistance to DNase I than lyophilized pK42-DNA particles. This study suggests that modification of pK42 with pLL-pEG diblock copolymers impacts polylysine's associative and binding behavior to DNA and resulting particle characteristics. Modulation of terpolymer composition in complexes can enable control over intracellular plasmid dissociation rates to improve transfection efficiency. PMID- 12643742 TI - 4-(2-aminooxyethoxy)-2-(ethylureido)quinoline-oligonucleotide conjugates: synthesis, binding interactions, and derivatization with peptides. AB - Oligo-2'-O-methylribonucleotides conjugated with 4-(2-aminooxyethoxy)-2 (ethylureido)quinoline (AOQ) and 4-ethoxy-2-(ethylureido)quinoline (EOQ) were prepared by reaction of the AOQ or EOQ phosphoramidite with the protected oligonucleotide on a controlled pore glass support. Deprotection with ethylenediamine enabled successful isolation and purification of the highly reactive AOQ-conjugated oligomer. Polyacrylamide gel electrophoresis mobility shift experiments showed that the dissociation constants of complexes formed between an AOQ- or EOQ-conjugated 8-mer and complementary RNA or 2'-O-methyl-RNA targets (9- and 10-mers) were in the low nM concentration range at 37 degrees C, whereas no binding was observed for the corresponding nonconjugated oligomer, even at a concentration of 500 nM. Fluorescence studies suggested that this enhanced affinity is most likely due to the ability of the quinoline ring of the AOQ or EOQ group to stack on the last base pair formed between the oligomer and target, thus stabilizing the duplex. The binding affinity of a 2'-O-methyl RNA 15 mer, which contained an alternating methylphosphonate/phosphodiester backbone, for a 59-nucleotide stem-loop HIV TAR RNA target, increased 2.3 times as a consequence of conjugation with EOQ. The aminooxy group of AOQ-conjugated oligomers is a highly reactive nucleophile, which reacts readily with aldehydes and ketones to form stable oxime derivatives. This feature was used to couple an AOQ-oligomer with leupeptin, a tripeptide that contains a C-terminus aldehyde group. A simple method was developed to introduce a ketone functionality into peptides that contain a cysteine residue by reacting the peptide with bromoacetone. The resulting keto-peptide was then coupled to the AOQ-oligomer. This procedure was used to prepare oligonucleotide conjugates of a tetrapeptide, RGDC, and a derivative of HIV tat peptide having a C-terminus cysteine. The combination of the unique reactivity of the aminooxy group and enhanced binding affinity conferred by its quinoline ring suggests that AOQ may serve as a useful platform for the preparation of novel oligonucleotide conjugates. PMID- 12643743 TI - N-succinimidyl 3-[(131)I]iodo-4-phosphonomethylbenzoate ([(131)I]SIPMB), a negatively charged substituent-bearing acylation agent for the radioiodination of peptides and mAbs. AB - An important criterion in design of acylation agents for the radioiodination of internalizing monoclonal antibodies (mAbs) is to maximize the retention of radioiodine in the tumor following mAb intracellular processing. We have previously shown that labeling methods that generate positively charged catabolites have enhanced tumor retention. Herein we have extended this strategy to investigate the potential utility of labeling internalizing mAbs with an acylation agent that yielded labeled catabolites that would be negatively charged at lysosomal pH. The negatively charged acylation agent, N-succinimidyl 3 [(131)I]iodo-4-phosphonomethylbenzoate ([(131)I]SIPMB), was prepared from its tin precursor, N-succinimidyl 4-di-tert-butylphosphonomethyl-3 trimethylstannylbenzoate (tBu-SPMTB), in 40% radiochemical yield. The free acid, 3-[(131)I]iodo-4-phosphonomethylbenzoic acid ([(131)I]IPMBA), was also prepared from the corresponding precursor, 4-di-tert-butylphosphonomethyl-3 trimethylstannylbenzoic acid (tBu-PMTBA), in 80% radiochemical yield. The rapidly internalizing mAb L8A4 was conjugated to [(131)I]SIPMB in 25-40% yield with preservation of its immunoreactivity. Internalization and processing in the U87DeltaEGFR glioma cell line was studied in a paired label format with L8A4 labeled with (125)I using the Iodogen method. Retention of initially bound radioactivity in these cells at 24 h from [(131)I]SIPMB-labeled mAb was approximately 6-fold higher than that for directly labeled mAb. Catabolite analysis demonstrated that this difference reflected an order of magnitude higher retention of low molecular weight species in these cells. The [(131)I]SIPMB-L8A4 conjugate was intact over the first 2 h; thereafter, lysine-[(131)I]SIPMB was the predominant catabolite. In contrast, L8A4 labeled using Iodogen rapidly gave rise to mono-[(125)I]iodotyrosine within 2 h, which then cleared rapidly from the cells. These results suggest that SIPMB could be a potent candidate for labeling internalizing mAbs and warrant further study. PMID- 12643744 TI - In vitro and in vivo gene transfer by an optimized alpha-cyclodextrin conjugate with polyamidoamine dendrimer. AB - The purpose of the present study is to optimize the structure of the polyamidoamine starburst dendrimer (dendrimer) conjugate with alpha-cyclodextrin (alpha-CDE conjugate) as a nonviral vector. alpha-CDE conjugates of dendrimer (generation 3, G3) with various average degrees of substitution (DS) of alpha-CyD of 1.1, 2.4, and 5.4 were prepared. alpha-CDE conjugates formed the complexes with pDNA, resulting in a change of the particle sizes of pDNA complexes, but the distinction of physicochemical properties among their vector/pDNA complexes was only very slight. The membrane-disruptive ability of alpha-CDE conjugates on liposomes encapsulating calcein and their cytotoxicity to NIH3T3 and HepG2 increased with an increase in the DS value of alpha-CyD. In vitro gene transfer activity of alpha-CDE conjugates in both NIH3T3 and HepG2 cells augmented as the charge ratio (vector/pDNA) increased, and the activity of alpha-CDE conjugate (DS 2.4) was the highest at higher charge ratios among dendrimer (G3), the three alpha-CDE conjugates, and TransFast. After intravenous administration of pDNA complexes in mice, alpha-CDE conjugate (DS 2.4) delivered pDNA more efficiently in spleen, liver, and kidney, compared with dendrimer and other alpha-CDE conjugates (DS 1.1 and 5.4). The potential use of alpha-CDE conjugate (G3, DS 2.4) could be expected as a nonviral vector in vitro and in vivo, and these data may be useful for design of alpha-CyD conjugates with other nonviral vectors. PMID- 12643745 TI - S-peptide as a potent peptidyl linker for protein cross-linking by microbial transglutaminase from Streptomyces mobaraensis. AB - We have found that ribonuclease S-peptide can work as a novel peptidyl substrate in protein cross-linking reactions catalyzed by microbial transglutaminase (MTG) from Streptomyces mobaraensis. Enhanced green fluorescent protein tethered to S peptide at its N-terminus (S-tag-EGFP) appeared to be efficiently cross-linked by MTG. As wild-type EGFP was not susceptible to cross-linking, the S-peptide moiety is likely to be responsible for the cross-linking. A site-directed mutation study assigned Gln15 in the S-peptide sequence as the sole acyl donor. Mass spectrometric analysis showed that two Lys residues (Lys5 and Lys11) in the S peptide sequence functioned as acyl acceptors. We also succeeded in direct monitoring of the cross-linking process by virtue of fluorescence resonance energy transfer (FRET) between S-tag-EGFP and its blue fluorescent color variant (S-tag-EBFP). The protein cross-linking was tunable by either engineering S peptide sequence or capping the S-peptide moiety with S-protein, the partner protein of S-peptide for the formation of ribonuclease A. The latter indicates that S-protein can be used as a specific inhibitor of S-peptide-directed protein cross-linking by MTG. The controllable protein cross-linking of S-peptide as a potent substrate of MTG will shed new light on biomolecule conjugation. PMID- 12643747 TI - Quantitative analysis of permeation peptide complexes labeled with Technetium 99m: chiral and sequence-specific effects on net cell uptake. AB - This study investigated sequence-specific cell uptake characteristics of Tat basic domain and related permeation peptides with an emphasis on residue chirality, length, and modified side chains. Effects on cell permeation of defined basic domain sequences within a library of 42 different peptides were evaluated using transport of radiolabeled peptides into human Jurkat leukemia cells. All other factors being equal, when the chirality of the peptide sequence was changed from l to d, uptake values increased up to 13-fold. Control experiments showed that the quantitative difference in uptake could not be attributed to increased decomposition of an l- versus a d-peptide by cellular or serum proteases. Furthermore, length, sequence, and type of chelation domain impacted peptide uptake into cells. The highest level of uptake was found with the following peptides: (23) d-Tat-Orn [Ac-rkkrr-orn-rrr-AHA-kgc-amide] and (33) d-poly-Arg(9) [Ac-rrrrrrrrr-AHA-kgc-amide]. The best of these peptide sequences could be employed as in vivo imaging and drug delivery agents to translocate substrates into cells. PMID- 12643746 TI - Novel galactosylated polyamine bolaamphiphiles for gene delivery. AB - We describe the synthesis of a series of alpha-galacto-omega-polyamine double chain bolaamphiphiles (Gal-CL) and report on the gene transfer mediated with lipoplexes they form either when used in conjunction with DOPE or with pcTG90:DOPE. Lipofection was investigated with human HepG2 and murine BNL-CL2 hepatocytes expressing the asialoglycoprotein (ASGP) receptor which displays a high affinity for galactosyl residues, and with A549 cells which do not express ASGP. Our results show that cationic N/P = 5 and 2.5 Gal-CL lipoplexes constitute very efficient nonspecific gene transfer systems. Lipofection experiments performed in the presence of asialofetuin (a high affinity ligand of ASGP) led us to evidence also the involvement of a specific receptor-mediated endocytosis pathway for the transfection of the ASGP(+) HepG2 or BNL-CL2 hepatocytes with some Gal-CL formulations. This work suggests that targetable lipopolyamines presenting a single galactose residue appear as promising synthetic vectors for specific gene delivery to ASGP(+) cells. PMID- 12643748 TI - Maleimide-oligo(ethylene glycol) derivatives of camptothecin as albumin-binding prodrugs: synthesis and antitumor efficacy. AB - In situ binding of thiol-reactive prodrugs to the cysteine-34 position of circulating albumin is a new approach in drug delivery. Therefore, five maleimide bearing derivatives of the anticancer drug camptothecin (CPT) were developed as albumin-binding prodrugs. These compounds were synthesized by reacting heterobifunctional cross-linkers based on oligo(ethylene glycols) [3-6 (O-CH(2) CH(2)) units] bearing a maleimide group on one end and a carboxylic acid group on the other with camptothecin 20-O-glycinate. Incorporating oligo(ethylene glycol) chains into the prodrugs enhanced their water-solubility when compared to the parent compound (up to 27-fold). HPLC studies showed that the prodrugs react almost quantitatively with the cysteine-34 position of endogenous albumin within a few minutes after incubation of the CPT derivatives with human blood plasma. The therapeutic potential of two of the prodrugs was assessed in nude mice bearing a colon xenograft (HT-29). Both albumin-binding derivatives of camptothecin were well-tolerated and showed enhanced antitumor efficacy when compared to CPT. PMID- 12643749 TI - Macromolecular MRI contrast agents with small dendrimers: pharmacokinetic differences between sizes and cores. AB - Large macromolecular MRI contrast agents with albumin or dendrimer cores are useful for imaging blood vessels. However, their prolonged retention is a major limitation for clinical use. Although smaller dendrimer-based MRI contrast agents are more quickly excreted by the kidneys, they are also able to visualize vascular structures better than Gd-DTPA due to less extravasation. Additionally, unlike Gd-DTPA, they transiently accumulate in renal tubules and thus also can be used to visualize renal structural and functional damage. However, these dendrimer agents are retained in the body for a prolonged time. The purpose of this study was to obtain information from which a macromolecular dendrimer-based MRI contrast agents feasible for use in further clinical studies could be chosen. Six small dendrimer-based MRI contrast agents were synthesized, and their pharmacokinetics, whole-body retention, and dynamic MRI were evaluated in mice to determine an optimal agent in comparison to Gd-[DTPA]-dimeglumine. Diaminobutane (DAB) dendrimer-based agents cleared more rapidly from the body than polyamidoamine (PAMAM) dendrimer-based agents with the same numbers of branches. Smaller dendrimer conjugates were more rapidly excreted from the body than the larger dendrimer conjugates. Since PAMAM-G2, DAB-G3, and DAB-G2 dendrimer-based contrast agents showed relatively rapid excretion, these three conjugates might be acceptable for use in further clinical applications. PMID- 12643750 TI - Controlled release of proteins from their poly(ethylene glycol) conjugates: drug delivery systems employing 1,6-elimination. AB - Several tripartate releasable PEG linkers (rPEG) that can provide anchimeric assistance to hydrolysis (cyclization prodrugs) were prepared and, after conjugation to lysozyme demonstrated rapid cleavage in rat plasma compared to nonassisted, permanently bound PEG. By varying the chemical structure and adding steric hindrance, the half-life of the protein conjugates can be adjusted from slow to very fast. The pharmacokinetics (PK) of regeneration of native protein, from various rPEG conjugates can, for the first time, be easily followed in the rat using green fluorescent protein. The PK in mice was also determined for rPEG Interleukin 2 (rPEG-IL-2) conjugates in vivo using an ELISA assay. Thus, a systematic study of rPEGylated proteins, either in vivo or in vitro during processing, has been investigated based on regeneration of native protein. The employment of releasable PEG polymers substantially broadens the applications of PEGylation drug delivery technology by introducing the benefits of controlled release of native protein therapeutics. PMID- 12643751 TI - Specific association of thiamine-coated gadolinium nanoparticles with human breast cancer cells expressing thiamine transporters. AB - Thiamine (vitamin B(1)) was investigated as a tumor-specific ligand for gadolinium nanoparticles. Solid nanoparticles containing gadolinium hexanedione (1.5 mg/mL) were engineered from oil-in-water microemulsion templates and coated with thiamine ligands. Thiamine ligands were synthesized by conjugating thiamine to either distearoylphosphatidylethanolamine (DSPE) or fluorescein via a poly(ethylene glycol) (PEG) spacer (Mw 3350). The efficiency of thiamine ligand attachment to nanoparticles was evaluated using gel permeation chromatography (GPC). Cell association studies were carried using a methotrexate-resistant breast cancer cell line, MTX(R)ZR75, transfected with thiamine transporter genes (THTR1 and THTR2). Thiamine-coated nanoparticle association with THTR1 and THTR2 cells was significantly greater than that with control breast cancer cells (MTX(R)ZR75 transfected with the empty expression vector pREP4) (p < 0.01; t test). The nanoparticle cell association was significantly dependent on the extent of thiamine ligand coating on nanoparticles, expression of thiamine transporters in cells, temperature of incubation, and the concentration of competitive inhibitor (free thiamine). Further studies are warranted to assess the potential of the engineered thiamine-coated gadolinium (Gd) nanoparticles in neutron capture therapy of tumors. PMID- 12643752 TI - Bioinspired pH-responsive polymers for the intracellular delivery of biomolecular drugs. AB - The biotechnology and pharmaceutical industries have developed a wide variety of potential therapeutics based on the molecules of biology: DNA, RNA, and proteins. While these therapeutics have tremendous potential, effectively formulating and delivering them have also been a widely recognized challenge. A variety of viruses and toxins have evolved multi-functional biomolecules to solve this problem by directing cellular uptake and enhancing biomolecular transport to the cytoplasm from the low pH endosomal compartment. In the study reported here, we have designed and synthesized bio-inspired, pH-responsive polymeric carriers, which we call "encrypted polymers", that mimic the multi-functional design of biology. These encrypted polymers target and direct cellular uptake, as well as enhance cytosolic delivery by disrupting endosomal membranes in a pH-dependent fashion. We show that the encrypted polymeric carriers significantly enhance the delivery of oligonucleotides and peptides to the cytoplasm of cultured macrophages, demonstrating the potential of this approach for delivery of biotherapeutics and vaccines. PMID- 12643753 TI - Low-pH-sensitive PEG-stabilized plasmid-lipid nanoparticles: preparation and characterization. AB - The acid-labile poly(ethyleneglycol)-diorthoester-distearoylglycerol lipid (POD), was used with a cationic lipid-phosphatidylethanolamine mixture to prepare stabilized plasmid-lipid nanoparticles (POD SPLP) that could mediate gene transfer in vitro by a pH triggered escape from the endosome. Nanoparticles of 60 nm diameter were prepared at pH 8.5 using a detergent dialysis method. The DNA encapsulation efficiency in the nanoparticles was optimal between 10 and 13 mol % ratio of cationic lipid and at a POD content of 20 mol %. The apparent zeta potential of the nanoparticles at 1 mM salt and pH 7.5 was positive, indicating cationic lipid on the external surface. However, the external layer of the nanoparticles was depleted in the cationic component compared to the starting mole ratio. Low pH sensitivity of the POD SPLP was characterized by a lag phase followed by a rapid collapse; at pH 5.3 the nanoparticles collapsed in 100 min. Nanoparticles prepared from a pH-insensitive PEG-lipid, PEG-distearoylglycerol had similar physicochemical characteristics as the POD SPLP but did not collapse at low pH. The POD SPLP had up to 3 orders of magnitude greater gene transfer activity than did the pH-insensitive nanoparticles. Both the pH-sensitive and pH insensitive nanoparticles were internalized to a qualitatively similar extent in a punctate pattern into cultured cells within 2 h of incubation with the cells; thus, increased gene transfer of the POD SPLP was due to a more rapid escape from the endosome rather than to greater cell association of these nanoparticles. These results suggest that the pH-sensitive stabilized plasmid-lipid nanoparticles may be a useful component of a synthetic vector for parenterally administered gene therapy. PMID- 12643754 TI - Alpha-oxo semicarbazone peptide or oligodeoxynucleotide microarrays. AB - We describe in this paper the preparation and characterization of semicarbazide glass slides and their use for the fabrication of microarrays using site-specific alpha-oxo semicarbazone ligation. The functional density and homogeneity of the semicarbazide glass slides were optimized by analyzing the reactivity of the layer toward a synthetic glyoxylyl fluorescent probe. Oligonucleotide microarrays were prepared by site-specific immobilization of glyoxylyl oligodeoxynucleotides. The slides were directly used in the hybridization assays using fluorescence detection and displayed a significant gain in sensibility as compared to the aldehyde glass slide/amino oligodeoxynucleotide chemistry. Semicarbazide slides were also used for the immobilization of a biotinylated peptide alpha-oxo aldehyde. The peptide microarrays allowed model interaction studies with streptavidin or an anti-biotin antibody. PMID- 12643755 TI - Generation of a functional monomolecular protein lattice consisting of an s-layer fusion protein comprising the variable domain of a camel heavy chain antibody. AB - Crystalline bacterial cell surface layer (S-layer) proteins are composed of a single protein or glycoprotein species. Isolated S-layer subunits frequently recrystallize into monomolecular protein lattices on various types of solid supports. For generating a functional protein lattice, a chimeric protein was constructed, which comprised the secondary cell wall polymer-binding region and the self-assembly domain of the S-layer protein SbpA from Bacillus sphaericus CCM 2177, and a single variable region of a heavy chain camel antibody (cAb-Lys3) recognizing lysozyme as antigen. For construction of the S-layer fusion protein, the 3'-end of the sequence encoding the C-terminally truncated form rSbpA(31)( )(1068) was fused via a short linker to the 5'-end of the sequence encoding cAb Lys3. The functionality of the fused cAb-Lys3 in the S-layer fusion protein was proved by surface plasmon resonance measurements. Dot blot assays revealed that the accessibility of the fused functional sequence for the antigen was independent of the use of soluble or assembled S-layer fusion protein. Recrystallization of the S-layer fusion protein into the square lattice structure was observed on peptidoglycan-containing sacculi of B. sphaericus CCM 2177, on polystyrene or on gold chips precoated with thiolated secondary cell wall polymer, which is the natural anchoring molecule for the S-layer protein in the bacterial cell wall. Thereby, the fused cAb-Lys3 remained located on the outer S layer surface and accessible for lysozyme binding. Together with solid supports precoated with secondary cell wall polymers, S-layer fusion proteins comprising rSbpA(31)(-)(1068) and cAbs directed against various antigens shall be exploited for building up monomolecular functional protein lattices as required for applications in nanobiotechnology. PMID- 12643756 TI - Differential gene expression profile between PC-14 cells treated with free cisplatin and cisplatin-incorporated polymeric micelles. AB - Cisplatin (CDDP)-incorporated polymeric micelles (CDDP/m) are a macromolecular carrier system possessing a time-modulated decaying property accompanied by sustained release of free drug. The gene expression profiles in nonsmall cell lung cancer PC-14 cells treated with free CDDP and CDDP/m were evaluated by a cDNA expression array for 807 genes. Although the total gene expression profile of the cells treated with CDDP/m approximated that of free CDDP in the hierarchical clustering analysis, a number of genes showed differential expression according to whether the cells had been treated with CDDP or CDDP/m. Ultimately, 50 genes with significant differential expression between cells treated with CDDP and CDDP/m were selected by principal component (PC) analysis and the unpaired t-test. The genes selected, including genes related to cell cycle regulation, apoptosis-related proteins, detoxification, and DNA repair enzymes, were considered to be related to CDDP-induced cytotoxicity. Interestingly, CDDP/m down-regulated the genes encoding integrins and matrix metalloproteinases (MMPs), which play an integral role in tumor invasion, metastasis, and angiogenesis, whereas free CDDP up-regulated them. The results suggest that use of the macromolecular carriers may yield additional therapeutic effects over free drug. PMID- 12643757 TI - N-Ac-DEVD-N'-(Polyfluorobenzoyl)-R110: novel cell-permeable fluorogenic caspase substrates for the detection of caspase activity and apoptosis. AB - N-Pentafluorobenzoyl-R110 (1a) and N-(2,3,4,5-tetrafluorobenzoyl)-R110 (1b) with enhanced cell retention properties, were prepared from rhodamine 110 (R-110) and the corresponding polyfluorobenzoyl chloride. N-Ac-DEVD-N'-pentafluorobenzoyl R110 (3a) and N-Ac-DEVD-N'-(2,3,4,5-tetrafluorobenzoyl)-R110 (3b) were prepared as tetrapeptide substrates for caspases. Substrate 3b was efficiently cleaved by human recombinant caspase-3 in an enzyme assay. Substrate 3b also was efficiently cleaved in cell-based apoptosis assays. After cleavage in apoptotic cells by activated caspases, the substrate becomes fluorescent as measured by flow cytometry. These substrates should prove useful in cell-based assays for studying apoptosis inducers and inhibitors. PMID- 12643758 TI - Site-specific PEGylation of hemoglobin at Cys-93(beta): correlation between the colligative properties of the PEGylated protein and the length of the conjugated PEG chain. AB - Increasing the molecular size of acellular hemoglobin (Hb) has been proposed as an approach to reduce its undesirable vasoactive properties. The finding that bovine Hb surface decorated with about 10 copies of PEG5K per tetramer is vasoactive provides support for this concept. The PEGylated bovine Hb has a strikingly larger molecular radius than HbA (1). The colligative properties of the PEGylated bovine Hb are distinct from those of HbA and even polymerized Hb, suggesting a role for the colligative properties of PEGylated Hb in neutralizing the vasoactivity of acellular Hb. To correlate the colligative properties of surface-decorated Hb with the mass of the PEG attached and also its vasoactivity, we have developed a new maleimide-based protocol for the site-specific conjugation of PEG to Hb, taking advantage of the unusually high reactivity of Cys-93(beta) of oxy HbA and the high reactivity of the maleimide to protein thiols. PEG chains of 5, 10, and 20 kDa have been functionalized at one of their hydroxyl groups with a maleidophenyl moiety through a carbamate linkage and used to conjugate the PEG chains at the beta-93 Cys of HbA to generate PEGylated Hbs carrying two copies of PEG (of varying chain length) per tetramer. Homogeneous preparations of (SP-PEG5K)(2)-HbA, (SP-PEG10K)(2)-HbA, and (SP-PEG20K)(2)-HbA have been isolated by ion exchange chromatography. The oxygen affinity of Hb is increased slightly on PEGylation, but the length of the PEG-chain had very little additional influence on the O(2) affinity. Both the hydrodynamic volume and the molecular radius of the Hb increased on surface decoration with PEG and exhibited a linear correlation with the mass of the PEG chain attached. On the other hand, both the viscosity and the colloidal osmotic pressure (COP) of the PEGylated Hbs exhibited an exponential increase with the increase in PEG chain length. In contrast to the molecular volume, viscosity, and COP, the vasoactivity of the PEGylated Hbs was not a direct correlate of the PEG chain length. There appeared to be a threshold for the PEG chain length beyond which the protection against vasoactivity is decreased. These results suggest that the modulation of the vasoactivity of Hb by PEG could be a function of the surface shielding afforded by the PEG, the latter being a function of the disposition of the PEG chain on the protein surface, which in turn is a function of the length of the PEG chain. Thus, the biochemically homogeneous PEGylated Hbs described in the present study, surface-decorated with PEG chains of appropriate size, could serve as potential candidates for Hb-based oxygen carriers. PMID- 12643759 TI - Novel intracellular delivery system of antisense oligonucleotide by self assembled hybrid micelles composed of DNA/PEG conjugate and cationic fusogenic peptide. AB - An antisense oligonucleotide (ODN), c-myb, was covalently conjugated to poly(ethylene glycol) (PEG) via an acid-cleavable phosphoramidate linkage to form a diblock copolymer-like structure. The phosphoramidate linkage between ODN and PEG was completely cleaved within 5 h in an endosomal acidic condition (pH 4.7). When complexed with a cationic fusogenic peptide, KALA, the ODN/PEG conjugate self-associated to form polyelectrolyte complex micelles in an aqueous solution. The anionic ODN segments were ionically interacted with cationic KALA peptide to form an inner polyelectrolyte complex core, while the PEG segments constituted a surrounding corona. Effective hydrodynamic volume of the micelles was ca. 70 nm with a very narrow size distribution. The polyelectrolyte complex micelles, composed of c-myb ODN-PEG conjugate and KALA, were transported into cells far more efficiently than c-myb ODN itself. They also exhibited higher antiproliferative activity against smooth muscle cells. This study demonstrates that the DNA/PEG hybrid micelles system can be applied for the delivery of antisense oligonucleotide. PMID- 12643760 TI - Mutants of immunotoxin anti-Tac(dsFv)-PE38 with variable number of lysine residues as candidates for site-specific chemical modification. 1. Properties of mutant molecules. AB - Chemical modification of proteins with substances such as poly(ethylene glycol) can add useful properties to proteins. Currently PEGylation is done in a random manner utilizing amino residues dispersed throughout a protein. For proteins such as immunotoxins, which have several different functional domains, random modification leads to inactivation. To determine if we could produce an immunotoxin with a diminished number of lysine residues so that chemical modification could be restricted to certain regions of the protein, we chose the recombinant immunotoxin anti-Tac(dsFv)-PE38 that has 13 lysine residues in the Fv portion and 3 in the toxin. We prepared a series of mutants with 0-12 lysines in the Fv and 0 or 3 in the toxin. Almost all of these molecules retain full biological activity. Our data indicate that replacement of lysine residues can be achieve without loss of biological potency. These molecules are a useful starting point to carry out site-specific PEGylation experiments. PMID- 12643761 TI - Synthesis and characterization of covalently linked single-stranded DNA oligonucleotide-dendron conjugates. AB - A solution-phase synthesis and characterization of covalent DNA-dendron conjugates is presented. Thiol-terminated 12-base oligonucleotides were added to second- and third-generation triazine-based dendrons via thiol/disulfide exchange chemistry. Single-stranded DNA oligonucleotides were successfully attached to dendrons at the core, the periphery, and both. Proof of structure for these architectures is derived primarily from mass spectrometry and polyacrylamide gel electrophoresis and complemented by labeling analysis using Ellman's reagent and degradation analysis using a reducing agent. PMID- 12643762 TI - Simultaneous lipidation of a characterized peptide mixture by chemoselective ligation. AB - The modification of a peptide antigen by a fatty acid such as palmitic acid is now recognized as a mean to induce cellular responses. Mixtures of lipopeptides, obtained by combining individually synthesized compounds, were shown to be promising synthetic vaccine candidates. Usually, in lipopeptide synthesis, the fatty acyl moiety is introduced on the crude peptide chain using solid-phase methods. The separation of the target compound from impurities by RP-HPLC is often complicated by the amphiphilic properties of lipopeptides and results in low overall yields. To overcome the difficulties associated with lipopeptide synthesis and mixture preparation, we have developed a method where the fatty acyl moiety is site-specifically and collectively introduced in solution onto a mixture of individually prepurified peptides. The lipidation is based on the quasistoichiometric and high-yielding ligation of a glyoxylyl lipid with hydrazinoacetyl peptides. The hydrazone constructs were prepared in a salt-free medium and could be isolated by direct lyophilization of the reaction mixture. This process is compatible with cysteinyl peptides, and no aggregation nor degradation could be observed. PMID- 12643763 TI - Synthesis and in vivo biodisposition of [14C]-quaternary ammonium-melphalan conjugate, a potential cartilage-targeted alkylating drug. AB - For the purpose of developing more selective anticancer drugs that would concentrate in the malignant cartilaginous tumors (chondrosarcomas), and so improve therapeutic index through a reduction of side effects, a quaternary ammonium (QA) conjugate of melphalan was synthesized and labeled with (14)C by linking the QA moiety to nitrogen mustard via an amide bond. Comparative pharmacokinetic study of [(14)C]-melphalan and its [(14)C]-QA conjugate conducted on rats showed that the two compounds were principally excreted by the urinary way. The blood elimination of the QA conjugate was faster than that of the melphalan. In the other hand a higher rate of radioactivity derived of [(14)C] MQA was found in feces. In the biodisposition for most organs, no striking differences were found between melphalan and its QA conjugate except for cartilages which exhibited more higher radioactivity level. Amounts of radioactivity derived from [(14)C]-QA conjugates measured in cartilaginous tissues until 1 h after injection demonstrate that the introduction of a QA moiety on melphalan allows the molecule to be carried selectively to cartilaginous tissues. As the [(14)C]-QA conjugate is radiolabeled on the chloroethyl alkylating moiety, levels of radioactivity measured in the cartilaginous tissues results from unchanged compound or metabolite having kept the active group. PMID- 12643764 TI - Molecular dynamics simulations of the unfolding of the starch binding domain from Aspergillus niger glucoamylase. AB - The 600 ps molecular dynamics simulations to investigate the unfolding of the starch binding domain from Aspergillus niger glucoamylase were conducted in vacuum as well as in an external field with the dielectric constant of 80 with temperature jump technique. Electrostatic interactions play an important role in determining the stability of the beta-strands in this domain. The starch binding site 1 is less stable than site 2 since it is more exposed to the surface. The disulfide bond between C509 and C604 is unstable since these two residues are located near the flexible linker domain and in the mobile loop region between beta-strands 6 and 7, respectively. The melting temperature, at which the total residual beta-strand content is 50% that of the solution structure, is about 544K for the simulations with dielectric constant of 80, leading to the estimated unfolding timescale of 0.48 ms in vitro. In addition, the unfolding of the starch binding domain is proposed to initiate from the interior region by the lost of the integrity of the secondary structure. PMID- 12643765 TI - Comparative analysis of docking motifs in MAP-kinases and nuclear receptors. AB - Nuclear receptor (NR) agonists induce activation of mitogen-activated protein kinases (MAPK) through an yet unknown rapid non-genomic mechanism. Vice versa, NR are targets for phosphorylation by MAPK. By multiple alignment of the amino acid sequences and comparative analysis of the secondary and tertiary structures we identified four peptides in MAPK with similarity to bona fide protein-protein interaction motifs in NR. In both molecule species, these motifs mediate selective docking to dimerization partners, coregulators or phosphoacceptors. We therefore propose that similar motifs may direct the site-specific association of NR with MAPK. Based on mutual allosteric interactions within a kinase-receptor complex, we discuss a novel principle how NR-agonists may regulate kinase activity and thus expression of hormone-dependent genes. PMID- 12643766 TI - A peptide derived from LFA-1 protein that modulates T-cell adhesion binds to soluble ICAM-1 protein. AB - Leukocyte function associated antigen 1 (LFA-1) and intercellular adhesion molecule 1 (ICAM-1) have been shown to be critical for adhesion process and immune response. Modulation or inhibition of the interaction between LFA-1/ICAM-1 interactions can result in therapeutic effects. Our group and others have shown that peptides derived from ICAM-1 or LFA-1 inhibit adhesion in a homotypic T-cell adhesion assay. It is likely that the peptides derived from ICAM-1 bind to LFA-1 and peptides derived from LFA-1 bind to ICAM-1 and inhibit the adhesion interaction. However, there are no concrete experimental evidence to show that peptides bind to either LFA-1 or ICAM-1 and inhibit the adhesion. Using NMR, CD and docking studies we have shown that an LFA-1 derived peptide binds to soluble ICAM-1. Docking studies using "autodock" resulted in LFA-1 peptide interacting with the ICAM-1 protein near Glu34. The proposed model based on our experimental data indicated that the LFA-1 peptide interacts with the protein via three intermolecular hydrogen bonds. Hydrophobic interactions also play a role in stabilizing the complex. PMID- 12643767 TI - The structural basis of a conserved P2 threonine in canonical serine proteinase inhibitors. AB - Bowman-Birk inhibitors (BBIs) are a well-studied family of canonical inhibitor proteins of serine proteinases. In nature, the active region of BBIs possesses a highly conserved Thr at the P2 position. The importance of this residue has been reemphasized by synthetic BBI reactive site loop proteinomimetics. In particular, this residue was exclusively identified for active chymotrypsin inhibitors selected from a BBI template-assisted combinatorial peptide library. A further kinetic analysis of 26 P2 variant peptides revealed that Thr provides both optimal binding affinity and optimal resistance against enzymatic turnover by chymotrypsin. Herein, we report the (1)H-NMR spectroscopic study of a 5-membered sub-set of these reactive site loop peptides representing a stepwise elimination of the Thr side-chain functionalities and inversion of its side-chain chirality. The P2 Thr variant adopts a three-dimensional structure that closely mimics the one of the corresponding region of the complete protein. This validates the use of this template for the investigation of structure-function relationships. While the overall backbone geometry is similar in all studied variants, conformational changes induced by the modification of the P2 side chain have now been identified and provide a rational explanation of the kinetically observed functional differences. Eliminating the gamma-methyl group has little structural effect, whereas the elimination of the gamma-oxygen atom or the inversion of the side chain chirality results in characteristic changes to the intramolecular hydrogen bond network. We conclude that the transannular hydrogen bond between the P2 Thr side-chain hydroxyl and the P5' backbone amide is an important conformational constraint and directs the hydrophobic contact of the P2 Thr side chain with the enzyme surface in a functionally optimal geometry, both in the proteinomimetic and the native protein. In at least four canonical inhibitor protein families similar structural arrangements for a conserved P2 Thr have been observed, which suggests an analogous functional role. Substitutions at P2 of the proteinomimetic also affect the conformational balance between cis and trans isomers at a distant Pro-Pro motif (P3'-P4'). Presented with a mixture of cis/trans isomers chymotrypsin appears to interact preferably with the conformer that retains the cis-P3' Pro-trans-P4' Pro geometry found in the parent BBI protein. PMID- 12643768 TI - A novel complexity measure for comparative analysis of protein sequences from complete genomes. AB - Analysis of sequence complexities of proteins is an important step in the characterization and classification of new genomes. A new measure has been proposed to compute sequence complexity in protein sequences based on linguistic complexity. The algorithm requires a single parameter, is computationally simple and provides a framework for comparative genomic analysis. Protein sequences were classified into groups of high or low complexity based on a quantitative measure termed F(c), which is proportional to the fraction of low complexity sequence present in the protein. The algorithm was tested on sequences of 196 non homologous proteins whose crystal structures are available at 5')-adenylyl-(3' --> 5')-adenosine [917-44-2] adenylyl-(3' --> 5') uridylyl-(3' --> 5')-guanosine [3494-35-7]. PMID- 12643776 TI - Commentary on IOM report: health insurance is a family matter. PMID- 12643777 TI - The joy of research. PMID- 12643778 TI - Reflections on the "So what?" dilemma. PMID- 12643779 TI - Unmet health care needs and impact on families with children with disabilities in Germany. AB - OBJECTIVES: We sought to determine the independent effect of unmet health needs on family burden, in addition to the effects of functional impairment and parental care load, in children and adolescents with disabilities. METHODS: We conducted a cross-sectional survey of 273 families with children with disabilities using ambulatory services at an academic children's hospital in Germany. We measured family burden using a translated version of the Impact on Family Scale (FABEL). Independent variables were unmet health needs in 4 areas (medical care, care coordination, health education, and psychosocial services), level of functional disability, and nursing care load at home. Control variables included the child's age and gender, maternal employment status, and parental educational attainment. RESULTS: Most children had complex health conditions such as brain injury, congenital malformations, metabolic disease, myopathies, and brain tumors. Nearly half of families (44.6%) received home nursing cash benefits, indicating high care load. Parents reported most unmet needs in the areas of psychosocial counseling (17.2%) and care coordination (8.1%). After controlling for sociodemographic factors, unmet health needs predicted family burden independently of type (mental retardation or mobility impairment) and number of disabilities and nursing care load. Although only a few parents reported lack of medical services, this factor also contributed significantly to family burden. Multivariate analysis with these variables explained 45% of the variance in impact on the family. CONCLUSIONS: Addressing unmet health needs may alleviate the impact of caring for a child with a disability. Further studies are needed to show more definitively that families can benefit from integrated services including psychosocial counseling. PMID- 12643780 TI - Continuity of care is associated with well-coordinated care. AB - CONTEXT: The importance of continuity of care as a means to promote care coordination remains controversial. OBJECTIVE: To determine if there is an association between having an objective measure of continuity of care and parental perception that care is well coordinated. DESIGN: Cross-sectional study. SETTING AND POPULATION: Seven hundred fifty-nine patients presenting to a primary care clinic completed surveys that included 5 items from the Components of Primary Care Index (CPCI) that relate to care coordination. MAIN PREDICTOR VARIABLE: A continuity of care index (COC) that quantifies the degree of dispersion of care among providers. MAIN OUTCOME MEASURES: Likelihood of parents reporting high scores on the care coordination domain as well as each of the 5 individual CPCI items related to care coordination. RESULTS: Greater continuity of care was associated with higher scores on the CPCI care-coordination domain (P <.001). Continuity of care was also specifically associated with increased odds of agreeing with all 5 individual CPCI items, including reporting that their child's provider "always knows about care my child received in other places" (OR 3.97 [2.11-7.49]), "communicates with the other health care providers my child sees" (OR 2.98 [1.63-5.44]), "knows the results of my child's visits to other doctors" (OR 2.02 [1.08-3.80]), and "always follows up on a problem my child has had, either at the next visit or by phone" (OR 6.20 [2.88-13.35]) and wanting one provider to coordinate all of the health care that the child receives (OR 3.28 [1.48-7.27]). CONCLUSIONS: Greater continuity of primary care is associated with better care coordination as perceived by parents. Efforts to improve and maintain continuity may be justified. PMID- 12643781 TI - Patterns of medical, educational, and mental health service use in a national sample of US children. AB - BACKGROUND: Special services are provided to children by the medical, educational, and mental health service systems, but it is unknown whether the same group of children receives different types of services or whether discrete subgroups predominantly receive these services. OBJECTIVE: To determine the proportions of children who receive special medical, educational, and mental health services, individually and in combination. DESIGN/METHODS: Using data from the 1994 National Health Interview Survey (NHIS) and its concurrent Disability Supplement (NHIS-D), we determined the proportions of children under age 18 (N = 30 032) who received each of 3 types of special services (medical, educational, mental health). We also examined the proportions of children who used combinations of these services. Finally, we determined whether the patterns of service use differed across age, sex, socioeconomic, or racial/ethnic groups. RESULTS: Overall, 9.6% of children used some type of special services. Somewhat greater proportions of children used specialized health services (5.5%) or educational services (5%) than mental health services (1.5%). About one third of children receiving services in each of the special medical and educational sectors are users of another type of services as well, but among children receiving mental health services, the majority also are recipients of another type of services. Highest rates of overall service use and of use in all 3 sectors were found among subgroups of children who were elementary school-aged or adolescents, male, of white non-Hispanic race/ethnicity, and living below the federal poverty level. This pattern is consistent with the literature on service utilization within the 3 systems of care and with the greater prevalence of children with chronic conditions in these same subgroups. CONCLUSION: These findings underscore the importance of communication in the planning of services for children across the medical, educational, and mental health sectors. Further research is needed to assess the actual degree of overlap among the various systems providing this care and the degree to which children and their families would benefit from more integrated systems. PMID- 12643782 TI - Experienced clinical educators improve their clinical teaching effectiveness. AB - OBJECTIVES: To determine if experienced clinical educators (CEs) can improve their teaching by incorporating 4 literature-based teaching methods into their instruction. METHODS: We trained 7 experienced CEs on the teaching methods during a monthly faculty development program. Each CE recorded use of these methods during 10 months on a personal digital assistant. We compared the CEs' teaching evaluations with those of nonparticipating faculty by analysis of variance at baseline, during the study period, and for 1 year after the study. RESULTS: Reported use of 2 teaching methods (priming and feedback) increased significantly over use at baseline; use of 2 other methods (teaching in the patient's presence and 1-2 focal teaching points) remained constant. Scores on the CEs' teaching evaluations were significantly higher during the study period on 1 item, whereas the comparison group showed no changes. The changes persisted during the follow up period. CEs reported that the teaching methods focused the learner and teacher, making subsequent encounters more productive. They also found that the act of entering data daily prompted them to reflect on their teaching. CONCLUSIONS: Experienced teachers can be persuaded to incorporate new methods into their daily teaching. Reflection on teaching is enhanced with group support and daily reminders. With these interventions, teaching effectiveness of these experienced educators improved. PMID- 12643783 TI - TV guides: exposure of hospitalized children to inappropriate programming. AB - OBJECTIVE: To determine if the policy recommendations of the American Academy of Pediatrics (AAP) regarding television viewing are heeded in an inpatient pediatric setting. SETTING: An inner-city academic medical center. PARTICIPANTS: Consecutive sample of 199 pediatric inpatients. METHODS: An investigator visited all pediatric inpatient rooms. Observations were made when the patient was present in the room, alert, and awake. The main outcome measure was exposure to inappropriate programming. In accordance with AAP guidelines, we used the established television rating system to code programs as appropriate versus inappropriate for children. We classified G- and PG-rated shows that centered on adult themes or that were not informational, educational, and nonviolent as inappropriate. We tested parental demographics, age of child, time of day, and presence of an adult in the room as possible predictors. RESULTS: Ninety-one percent of the patients had the television turned on. Of these, 53% were tuned to inappropriate programming. Exposure to inappropriate programming did not differ by parental ethnicity, language, or education. Exposure was highest for infants and toddlers as compared with children or adolescents (74% vs 40% vs 52%, P =.001); it did not vary by time of day. Exposure was higher when an adult was present in the room (58% vs 42%, P =.05); this effect was most pronounced for adolescents (68% vs 41%, P =.05). CONCLUSIONS: Despite AAP recommendations, hospitalized children are exposed to inappropriate programming, especially when parents are present in the room. Hospital stays may be a unique opportunity to educate parents about the AAP recommendations for television viewing by children. PMID- 12643784 TI - A potential pitfall in provider assessments of the quality of asthma control. AB - BACKGROUND: Pediatricians elicit information about asthma control from parents to help formulate management plans. If parents of children with significant asthma symptoms inappropriately indicate good control, physician recommendations may not be optimal. We examined whether a single general question about asthma control might lead to inaccurate assessment of severity. DESIGN/METHODS: Children 3-7 years of age who met the National Heart, Lung, and Blood Institute (NHLBI) criteria for mild persistent to severe asthma were identified from 40 urban schools. A phone survey of their parents provided demographic information, symptom frequency, medication use, and general interpretation of their child's asthma control. Chi-square analyses compared the parent's general interpretation of control with demographic characteristics and measures of asthma severity. RESULTS: One hundred sixty of 224 eligible children participated in this study. Seventy-eight percent were described as in good asthma control. General assessment of asthma control did not vary by demographic characteristics. Parents were as likely to describe children with daily asthma symptoms in good control as they were to describe children with less frequent symptoms in good control. Parents were less likely to report good control in children using daily rescue medications when compared with children with less frequent medication use (65% vs 82%, P =.03), but the majority of children in both groups was described as having good control. CONCLUSIONS: Most parents underestimated the severity of their child's asthma and reported good control with their global assessment. Parents frequently reported good control even when the children had daily asthma symptoms. Pediatricians should ask about specific asthma symptoms during patient encounters because a global question about asthma control likely will result in underestimations of asthma severity. PMID- 12643785 TI - Reported level of supervision of young children while in the bathtub. AB - BACKGROUND: Unintentional drownings cause over 600 deaths annually among children < or =5 years of age. Bathtubs are one of the leading sites for such drownings for children < or =2 years of age and especially for children 1 year or younger. OBJECTIVE: To determine reported levels of supervision of children while they are in the bathtub. METHODS: A face-to-face questionnaire was administered to a convenience sample of parents or guardians of children < or =5 years of age who presented to the emergency centers of 2 children's hospitals. The investigators developed a 17-point survey that included items related to general demographics and bathtub supervision. Parents and guardians were asked details concerning who supervised while the child was bathing, any unsupervised periods, length of unsupervised periods, and activities during any unsupervised periods. RESULTS: A total of 259 families participated in the survey (16% Caucasian, 79% African American, 1% Hispanic, 3% Asian, and 1% biracial). The mean caregiver age was 29.1 years (SD 7.9). The mean age of the child was 30.5 months (SD 22). Overall, 31% of the respondents reported leaving their child unsupervised for some period of time in the bathtub. The mean age of children left alone was 37.5 months, with the youngest being 5 months old. Seventeen percent of children < or =24 months of age (N = 123, mean age 12.7 months) and 15% of children < or =12 months of age (N = 60, mean age 6.7 months) were reportedly at times left unsupervised in the bathtub. Common caregiver activities when leaving the child unsupervised included getting a towel or diapers, answering a phone, and cooking. A 5-month-old child was left unsupervised for >2 minutes, and an 8-month-old child was reportedly left unsupervised for >5 minutes to cook a meal. In addition, 20 of 259 respondents (7.7%) reported that their children bathed alone before the age of 5 years, and 4 respondents (1.5%) reported that their children bathed alone before the age of 2 years. Although most primary supervisors were adults, 5 children were at times supervised by children <10 years of age. No significant differences were seen based on the hospital, race, or educational level of the families. CONCLUSIONS: Many parents reported leaving their young children at times inadequately supervised in the bathtub. This occurred in children as young as 5 months of age. Given the potential risk of drowning when their children are inadequately supervised, parents should be advised concerning proper supervision in early anticipatory guidance. PMID- 12643786 TI - The effort to make oral health a more visible issue to pediatricians. PMID- 12643789 TI - Dose-dependent differential expression of protein kinase C isozymes in mouse lymphocytes after gamma irradiation in vivo and ex vivo. AB - Protein kinase C (PKC, now known as Prkc) plays an important role in the response of cells to radiation, but little is known about the specific response of each isozyme in the radiation-induced response of cells in whole animals. However, most studies are based on single cells. There is a paucity of data on signaling after whole-body irradiation. In this study, a comparison has been made between the expression of Prkc isozymes after in vivo and ex vivo irradiation. There was a significant difference in the dose response of the isozymes. In animals in which lymphocytes were irradiated ex vivo, the expression of the Prkca isozyme was found to be maximum at 3 Gy, while in vivo irradiation did not increase the expression beyond that of 1 Gy. Prkcd was marginally activated after 0.1 Gy ex vivo irradiation, whereas there was significant activation of expression after in vivo irradiation with 3 Gy. The response of Prkcz was found to be similar to that of Prkcd. Prkc is a crucial enzyme that is being used to manipulate the response of tumors to radiotherapy. Conventional radiotherapy is delivered at low doses, and hence only those isozymes that are activated at these doses should be taken into consideration. Moreover, the differences between the response of a single cell and that of the whole animal must be considered. PMID- 12643788 TI - Epidermal growth factor and ionizing radiation up-regulate the DNA repair genes XRCC1 and ERCC1 in DU145 and LNCaP prostate carcinoma through MAPK signaling. AB - This work examined the importance of radiation-induced and ligand-induced EGFR ERK signaling for the regulation of DNA repair proteins XRCC1 and ERCC1 in prostate carcinoma cells, DU145 (TP53(mut)), displaying EGFR-TGFA-dependent autocrine growth and high MAPK (ERK1/2) activity, and LNCaP (TP53(wt)) cells expressing low constitutive levels of ERK1/2 activity. Using quantitative RT-PCR and Western analyses, we determined that ionizing radiation activated the DNA repair genes XRCC1 and ERCC1 in an ERK1/2-dependent fashion for each cell line. After irradiation, a rapid increase followed by a decrease in ERK1/2 activity preceded the increase in XRCC1/ERCC1 expression in DU145 cells, while only the rapid decrease in ERK1/2 preceded the increase in XRCC1/ERCC1 expression in LNCaP cells. Administration of EGF, however, markedly increased the up-regulation of phospho-ERK, ERCC1 and XRCC1 in both cell lines. Although the EGFR inhibitor tyrphostin (AG-1478) and the MEK inhibitor PD90859 both attenuated EGF-induced levels of the ERCC1 and XRCC1 protein, PD98059 blocked the induction of ERCC1 and XRCC1 by radiation more effectively in both cell lines. Inhibition of ERK at a level that reduced the up-regulation of DNA repair led to the persistence of apurinic/apyrimidinic (AP) sites of DNA damage and increased cell killing. Taken together, these data imply a complex control of DNA repair activation that may be more generally dependent on MAPK (ERK1/2) signaling than was previously noted. These data provide novel insights into the capacity of the EGFR-ERK signaling to modulate DNA repair in cancer cells and into the functional significance of this signaling. PMID- 12643790 TI - Mitochondrial DNA heteroplasmy in laboratory mice experimentally enclosed in the radioactive Chernobyl environment. AB - Mitochondrial DNA heteroplasmy using the protein-coding cytochrome b (Mtcyb) gene was assessed in laboratory mice (C57BL/6 and BALB/c) exposed to the Chernobyl environment. Subacute to subchronic (30-40 days) exposure resulted in a cumulative radiation dose of 1.2-1.6 Gy ( approximately 0.04 Gy/day). Mice were sampled prior to introduction into the enclosures and again after removal from the enclosures. Nucleotide variation (site heteroplasmy) in 306 pre-exposure Mtcyb gene copies (122400 base pairs) was compared to variation in 354 postexposure gene copies (141600 base pairs). Five mutant copies, each characterized by a single nucleotide substitution, were observed (four in the pre exposure samples, one in a postexposure sample). The frequencies of mutant gene copies and nucleotide substitutions in pre-exposure and postexposure samples were not significantly different. This suggests that this type of exposure (i.e. low dose rate) does not pose a significant mutation risk to the Mtcyb gene in digit tissue. Furthermore, no significant radiation risk to analogous human tissues may exist when occupational exposures involve low dose rates such as these. Finally, linear, cumulative models of genetic risk currently used to estimate radiation induced effects are likely to be inappropriate for low-dose-rate exposures and need to be re-evaluated critically. PMID- 12643791 TI - Possible role of nitric oxide in radiation-induced salivary gland dysfunction. AB - In this study, we developed a murine model of xerostomia to elucidate the mechanism of radiation-induced salivary gland dysfunction and determined the levels of nitric oxide (NO) in the salivary glands to assess its involvement in the salivary dysfunction induced by radiation. In addition, an inhibitor of NO synthesis was administered to the model in vivo, and its effect on saliva secretion was investigated. Salivary gland irradiation at a dose of 15 Gy caused a significant decrease in secretion compared to unirradiated salivary glands. There were no marked differences between the irradiated mice and unirradiated mice in water or food consumption or in body weight changes. The NO levels in the cultured salivary gland epithelial cells were increased by treatment with a combination of interferon gamma (Ifng), interleukin 1-beta (Il1b), and tumor necrosis factor alpha (Tnfa). Irradiation increased the NO level in the salivary gland tissue. The presence of N(G)-monomethyl-l-arginine acetate (l-NMMA), an inhibitor of NO synthesis, caused a decrease in the NO level in cultured salivary gland tissues after irradiation. Administration of l-NMMA to irradiated mice improved saliva secretion. These results suggest that excessive production of NO induced by radiation is involved in the formation of radiation-induced xerostomia. The finding that administration of an inhibitor of NO synthesis ameliorated the dysfunction of irradiated salivary glands indicates that NO plays a role as a mediator of the dry mouth symptoms that occur after irradiation. PMID- 12643792 TI - The cell membrane as a biosensor of oxidative stress induced by radiation exposure: a multiparameter investigation. AB - The role of biological membranes as a target in biological radiation damage remains unclear. The present study investigates how the biochemical and biophysical properties of a simple biological model, i.e. human erythrocyte membranes, are altered after exposure to relatively low doses of (60)Co gamma rays. Lipid peroxidation increased in the hours after radiation exposure, based on measurements of MDA and on the lipid peroxidation index after parinaric acid incorporation. Protein carbonyl content also increased rapidly after radiation exposure. An imbalance between the radiation-mediated oxidative damages and the antioxidant capacity of the erythrocytes was observed in the hours after radiation exposure. Antioxidant enzyme activities, mainly catalase and glutathione peroxidase, were found to decrease after irradiation. The development of a radiation-induced oxidative stress probably explains the reorganization of the fatty acid pattern 72 h after radiation exposure. The phosphatidylethanolamine (PE) fatty acids of the (n-3) and (n-6) series decreased, while the PE saturated fatty acid content increased. All these modifications may be involved in the variation of the biophysical properties of the membranes that we noted after radiation exposure. Specifically, we observed that the lipid compartment of the membrane became more fluid while the lipid protein membrane interface became more rigid. Taken together, these findings reinforce our understanding that the cell membrane is a significant biological target of radiation. Thus the role of the biological membrane in the expression and course of cell damage after radiation exposure must be considered. PMID- 12643794 TI - X-ray-induced DNA double-strand breaks in mouse l1210 cells: a new computational method for analyzing neutral filter elution data. AB - The aim of this article is to present a method for studying the shape of the dose and repair responses for X-ray-induced double-strand breaks (DSBs) as measured by neutral filter elution (NFE). The approach is closely related to a method we developed for the use of specific molecular size markers and used for determination of the absolute number of randomly distributed radiation-induced DSBs by pulsed-field gel electrophoresis (PFGE). Mouse leukemia L1210 cells were X-irradiated with 0-50 Gy. Samples were then evaluated both with PFGE and with NFE. Assuming that with both migration (PFGE) and elution (NFE), a heterogeneous population of double-stranded DNA fragments will start with the smallest fragments and proceed with increasingly larger fragments, it is possible to match the migration behavior of fractions of fragments smaller than a certain size to the fraction eluted at a specific time. This assumption does not exclude the possibility of DNA being sheared in the NFE filter. The yield, as determined by the size markers in PFGE, was used to find the corresponding elution times in the NFE experiment. These experimentally used elution times could then reversely be interpreted as size markers which finally were used to calculate DSBs/Mbp as a function of X-ray dose. The resulting lines were almost straight. The data were also plotted as relative elution and showed that, as expected, the dose response then appears with a more pronounced sigmoid shape. PMID- 12643793 TI - Radiation response of cells during altered protein thiol redox. AB - The major focus of this work was to investigate how altered protein thiol redox homeostasis affects radiation-induced cell death. We used the cells of wild-type CHO cell line K1, the CHO cell line E89, which is null for G6PD activity, and a radiation-sensitive CHO cell line, XRS5. The protein-thiol redox status of cells was altered with cell-permeable disulfides, hydroxyethyldisulfide (HEDS) or lipoate. HEDS is primarily reduced by thioltransferase (glutaredoxin), with GSH as the electron donor. In contrast, lipoate is reduced by thioredoxin reductase. HEDS was reduced at a greater rate than lipoate by G6PD-containing K1 (wild-type) cells. Reduction of disulfides by G6PD-deficient cells was significantly slower with HEDS as substrate and was nearly absent with lipoate. The rate of reduction of HEDS by E89 cells decelerated to near zero by 30 min, whereas the reduction continued at nearly the same rate during the entire measurement period for K1 cells. HEDS treatment decreased the GSH and protein thiol (PSH) content more in G6PD-deficient cells than in G6PD-containing cells. On the other hand, lipoate did not significantly alter the protein thiol, but it increased the GSH in K1 cells. Acute depletion of GSH by l-buthionine-sulfoximine (l-BSO) in combination with dimethylfumarate significantly decreased the rate of reduction of HEDS by K1 cells close to that of G6PD-deficient cells. Prior GSH depletion by l-BSO alone significantly decreased the PSH in glucose-depleted E89 cells exposed to HEDS, but this did not occur with K1 cells. The radiation response of G6PD-deficient cells was significantly sensitized by HEDS, but HEDS did not have this effect on K1 cells. The DNA repair-deficient XRS5 CHO cells displayed the same capacity as K1 cells for HEDS reduction, and like K1 cells the XRS5 cells were not sensitized to radiation by HEDS treatment. Deprivation of glucose, which provides the substrate for G6PD in the oxidative pentose phosphate cycle, decreased the rate of bioreduction of HEDS and lipoate in G6PD-containing cells to the level in G6PD deficient cells. In the absence of glucose, HEDS treatment diminished non-protein thiol and protein thiol to the same level as those in G6PD-deficient cells and sensitized the K1 cells to HEDS treatment. However, depletion of glucose did not alter the sensitivity of XRS5 cells in either the presence or absence of HEDS. Overall the results suggest a major role for pentose cycle control of protein redox state coupled to the activities of the thioltransferase and thioredoxin systems. The results also show that protein thiol status is a critical factor in cell survival after irradiation. PMID- 12643795 TI - Measurement of prompt DNA double-strand breaks in mammalian cells without including heat-labile sites: results for cells deficient in nonhomologous end joining. AB - Ionizing radiation induces prompt single-strand breaks and double-strand breaks in DNA. In addition, labile sites are induced that can be converted to breaks by heat or mild alkali. When such labile lesions are present within multiply damaged sites, additional double-strand breaks can form. Current protocols for measurement of DNA double-strand breaks involve a lysis step at an elevated temperature, and consequently breaks from heat-labile sites will be generated during lysis and will be included in the measurement. However, such sites may not develop into breaks within the cell and therefore may not need DNA double-strand break repair processes for elimination. We present here a new lysis and pulsed field gel electrophoresis protocol that is carried out entirely at 0-4 degrees C and thus avoids inclusion of heat-labile sites in the measurement. The new recommended lysis procedure involves two steps: The first step includes proteinase K, which has sufficient activity at 0 degrees C to support lysis, and the second step includes a high-salt buffer to further free the DNA from proteins and other cellular structures. Using various tests, we conclude that lysis is sufficient with this procedure to allow accurate determination of double-strand breaks by pulsed-field gel electrophoresis. Using the new protocol, it was found that heat-labile sites account for 30% of the initial number of double-strand breaks measured by conventional protocols after exposure to low-LET radiation. In addition, we show that heat-labile sites that can be converted to double-strand breaks are repaired with fast kinetics and are almost completely eliminated after 1 h at 37 degrees C. A study of cells deficient in nonhomologous end joining reveals that the residual fast repair response typically seen in such cells is solely due to repair at heat-labile sites and is not due to repair of prompt DSBs. PMID- 12643796 TI - Joint effects of radiation and smoking on lung cancer risk among atomic bomb survivors. AB - Results are given on the joint effect of radiation exposure and cigarette smoking on lung cancer risks among A-bomb survivors, based on 592 cases through 1994. Information on smoking was derived from mail surveys and clinical interviews of 45113 persons in the Radiation Effects Research Foundation cohort. Radiation and smoking effects on lung cancer are found to be significantly sub-multiplicative and quite consistent with additivity. The smoking relative risk, previously very low in studies of this cohort, is now similar to that found in Western populations. This increase is likely to be related to the scarcity of cigarettes during and after the war. The smoking relative risk depends little on sex. After adjusting for smoking, the radiation-related risks relative to background rates for nonsmokers are similar to those for other solid cancers: a sex-averaged ERR/Sv of about 0.9 with a female:male sex ratio of about 1.6. Adjusting for smoking removes a spuriously large female:male ratio in radiation relative risk due to confounding between sex and smoking level. The adjustment also removes an artifactual age-at-exposure effect in the radiation relative risk, opposite in direction to other cancers, which is due to birth cohort variation in lung cancer rates. PMID- 12643797 TI - Mortality after radiological investigation with radioactive Thorotrast: a follow up study of up to fifty years in Portugal. AB - Cerebral angiography using a radioactive radiological contrast medium, Thorotrast, was pioneered by Moniz in Portugal in the 1920s. Thorotrast is retained by the reticuloendothelial system, with a biological half-life of several hundred years, so that such patients suffer lifetime exposure to internal radiation. We studied mortality in Portuguese patients who were administered Thorotrast during the period 1928-1959 and in a comparison group of patients who received nonradioactive contrast agents. There were 1096 systemically exposed, 1014 unexposed, and, unique to the Portuguese study, 240 locally exposed Thorotrast patients who were successfully traced and followed up to the end of 1996. Mortality was significantly raised among systemically exposed Thorotrast patients relative to those unexposed for all causes [relative risk (RR) = 2.63], all neoplasms (RR = 6.72), liver cancer (RR = 42.4), chronic liver disease (RR = 5.12), other non-neoplastic diseases of the digestive system (RR = 4.87), neoplastic (RR = 21.9) and non-neoplastic hematological disorders (RR = 6.00), and non-neoplastic diseases of the respiratory system (RR = 4.31). Risks for most of these conditions increased significantly with time since first administration of the contrast medium and with cumulative alpha-particle radiation dose. Mortality was also significantly raised for non-neoplastic disorders of the nervous system (RR = 12.7) and ill-defined conditions (RR = 3.74), but these associations are likely to reflect the initial diagnosis, not Thorotrast exposure, because risks declined significantly with time and/or dose. There were no significant excess deaths from oropharyngeal or nasal cancers, or from any other cause, among patients exposed to Thorotrast locally for visualization of the perinasal sinuses, and no clear trend in risk with time since exposure. This study shows an association between systemic, but not local, exposure to Thorotrast and mortality from liver cancer, chronic liver disease, and neoplastic and non-neoplastic hematological disorders, with risks for these conditions remaining high for over 40 years after administration. Liver conditions, but not hematological disorders, showed a strong and consistent gradient with cumulative alpha-particle radiation dose. PMID- 12643798 TI - Analysis of dose distribution for heavily exposed workers in the first criticality accident in Japan. AB - The first criticality accident in Japan occurred in a uranium processing plant in Tokai-mura on September 30, 1999. The accident, which occurred while a large amount of enriched uranyl nitrate solution was being loaded into a tank, led to a chain reaction that continued for 20 h. Two workers who were pouring the uranium solution into the tank at the time were heterogeneously exposed to neutrons and gamma rays produced by nuclear fission. Analysis of dose distributions was essential for the understanding of the clinical course observed in the skin and organs of these workers. We developed a numerical simulation system, which consists of mathematical human models and Monte Carlo radiation transport programs, for analyzing dose distributions in various postures and applied the system to the dose analysis for the two workers. This analysis revealed the extreme heterogeneity of the doses from neutrons and gamma rays in the skin and body, which depended on the positions and postures of the workers. The detailed dose analysis presented here using color maps is indispensable for an understanding of the biological effects of high-dose exposure to a mixed field of neutrons and gamma rays as well as for the development of emergency treatments for victims of radiation exposure. PMID- 12643799 TI - Radiation-activated nuclease activity of o,o'-Diphenyleneiodonium cations (DPI): a reductively initiated chain reaction involving the C1' chemistry. AB - o,o'-Diphenyleneiodonium cations (DPI) convert relatively harmless radiation produced electrons into efficient DNA cleaving agents. The cleavage products are unaltered DNA bases, 5-methylenefuranone (5-MF), and a complete set of 3' and 5' phosphorylated DNA fragments. The production of alkali-labile sites is a minor factor in the process. Based on the production of 5-MF, it is concluded that DNA cleavage by DPI cations involves (but may not be limited to) the C1' chemistry. The loss of 3-aminoDPI (ADPI) cations bound to highly polymerized calf thymus DNA appears to be due to a short-chain reaction with an apparent length of up to 2.1 ADPI cations consumed for each radiation-produced electron. The suggested chain reaction mechanism includes the one-electron oxidation of DNA radicals (including the C1' sugar radical) by ADPI cations bound to the same duplex. The yields of DNA loss in complexes formed by ADPI with short synthetic duplexes indicate that there is more than a 60% probability of DNA damage after one-electron reduction of ADPI. PMID- 12643800 TI - Estimation of the SAR in the human head and body due to radiofrequency radiation exposure from handheld mobile phones with hands-free accessories. AB - It was reported by others that hands-free accessories increase the absorption of RF energy in a human head compared to a handset alone. The results of this study show that the opposite is observed when proper dosimetric methods are employed. It is pointed out that for correct estimation of the exposure level it is necessary to use appropriate physical and experimental models and measurement instrumentation, following internationally recommended standards. The human phantoms used for measurements involving the hands-free accessories should include the torso; i.e., measurements should not be performed on the head phantom alone. This has a significant impact on the results because the RF energy coupled into the leads of hands-free accessories is strongly attenuated by the body. Numerical simulations using the Finite-Difference Time-Domain (FDTD) method and experimental measurements with a miniature electric-field probe are in good agreement and show a decrease, not an increase, in RF energy exposure in the human head from hands-free accessories. PMID- 12643801 TI - Genotoxic potential of 1.6 GHz wireless communication signal: in vivo two-year bioassay. AB - Timed-pregnant Fischer 344 rats (from nineteenth day of gestation) and their nursing offspring (until weaning) were exposed to a far-field 1.6 GHz Iridium wireless communication signal for 2 h/day, 7 days/week. Far-field whole-body exposures were conducted with a field intensity of 0.43 mW/cm(2) and whole-body average specific absorption rate (SAR) of 0.036 to 0.077 W/kg (0.10 to 0.22 W/kg in the brain). This was followed by chronic, head-only exposures of male and female offspring to a near-field 1.6 GHz signal for 2 h/day, 5 days/week, over 2 years. Near-field exposures were conducted at an SAR of 0.16 or 1.6 W/kg in the brain. Concurrent sham-exposed and cage control rats were also included in the study. At the end of 2 years, all rats were necropsied. Bone marrow smears were examined for the extent of genotoxicity, assessed from the presence of micronuclei in polychromatic erythrocytes. The results indicated that the incidence of micronuclei/2000 polychromatic erythrocytes were not significantly different between 1.6 GHz-exposed, sham-exposed and cage control rats. The group mean frequencies were 5.6 +/- 1.8 (130 rats exposed to 1.6 GHz at 0.16 W/kg SAR), 5.4 +/- 1.5 (135 rats exposed to 1.6 GHz at 1.6 W/kg SAR), 5.6 +/- 1.7 (119 sham exposed rats), and 5.8 +/- 1.8 (100 cage control rats). In contrast, positive control rats treated with mitomycin C exhibited significantly elevated incidence of micronuclei/2000 polychromatic erythrocytes in bone marrow cells; the mean frequency was 38.2 +/- 7.0 (five rats). Thus there was no evidence for excess genotoxicity in rats that were chronically exposed to 1.6 GHz compared to sham exposed and cage controls. PMID- 12643802 TI - Australia has more than enough solar UV radiation. PMID- 12643803 TI - Should sunglasses be required to comply with the sunglass standard? PMID- 12643804 TI - Sunglasses and sunglass standards. AB - Sunglasses and sunglass standards are reviewed from the point of view of the history of sunglasses and the development of sunglass standards. The need for eye protection from solar radiation is discussed and the provisions of the various national sunglass standards are discussed in relation to that need. PMID- 12643805 TI - Knowledge of the ocular effects of diabetes among the general population of Australia and the members of Diabetes Australia. AB - BACKGROUND: Early diagnosis and treatment of diabetes mellitus is important for limiting its adverse effects. We sought to determine the level of awareness of diabetes and its ocular complications within the community and among the members of Diabetes Australia. METHODS: Two groups were surveyed: 1. 1,000 people randomly selected from the Queensland electoral role (people without diabetes) and 2. A random sample of 500 members of Diabetes Australia (Queensland) (people with diabetes). The surveys consisted primarily of questions relating to the person's current knowledge of diabetes, including ocular and health complications, and their knowledge of available eye care services. RESULTS: The rate of return was greater for the members of Diabetes Australia (58.6 per cent) than for the public (33.5 per cent). The majority of respondents without diabetes tended to consult an optometrist for their eye care (76 per cent), while the group with diabetes consulted ophthalmologists more (63.6 per cent). People with diabetes tended to have more frequent eye examinations, 86.7 per cent had been for an eye examination within the preceding 18 months (compared with 36.6 per cent of the public). The level of awareness of the ocular effects of diabetes was high: 96 per cent of people with and 78.5 per cent of people without diabetes knew that diabetes could be sight-threatening. CONCLUSION: The membership of Diabetes Australia had a greater understanding of the possible ocular effects of diabetes and the need for regular eye examinations than the general community. Potential sample bias and the fact that respondents could infer a link between eye problems and diabetes from the survey questions should be taken into account in interpreting the data presented here. PMID- 12643806 TI - The relationship between central corneal thickness and Goldmann applanation tonometry. AB - BACKGROUND: The aim of this study was to investigate the relationship between Goldmann applanation tonometry and central corneal thickness in a large sample of healthy eyes. METHOD: Five hundred eyes of 500 subjects (253 women, 50.6 per cent and 247 men, 49.4 per cent) were analysed in a prospective healthy population study. Mean age of the sample was 31 +/- 8 years. Goldmann applanation tonometry was carried out by one physician. Tonometric values were the mean of three consecutive readings. Subsequently, another physician carried out ultrasonic pachymetry with the DGH 2000 AP ultrasonic pachymeter (DGH Technology Inc, San Diego, USA). Ten measurements were made at the centre of the cornea of each eye. The lowest value was used for analysis. RESULTS: Applanation tonometry and central corneal thickness were correlated (r = 0.184, p < 0.001). There was no significant correlation between corneal thickness and age (r = 0.083, p = 0.065), mean spherical equivalent refraction (r = 0.083, p = 0.065) or visual acuity (r = 0.036, p = 0.187). CONCLUSION: In normal eyes, there is no statistically significant correlation between changes of intraocular pressure and changes of central corneal thickness but they suggest a relationship between intraocular pressure and central corneal thickness. Goldmann applanation tonometry has a systematic error in accuracy of intraocular pressure readings of healthy eyes caused by its dependence on central corneal thickness. Measurement of corneal thickness by optometrists should be the first step in diagnosing intraocular pressure pathologies. PMID- 12643807 TI - The Min Min light and the Fata Morgana. An optical account of a mysterious Australian phenomenon. AB - BACKGROUND: Despite intense interest in this mysterious Australian phenomenon, the Min Min light has never been explained in a satisfactory way. METHODS & RESULTS: An optical explanation of the Min Min light phenomenon is offered, based on a number of direct observations of the phenomenon, as well as a field demonstration, in the Channel Country of Western Queensland. This explanation is based on the inverted mirage or Fata Morgana, where light is refracted long distances over the horizon by the refractive index gradient that occurs in the layers of air during a temperature inversion. Both natural and man-made light sources can be involved, with the isolated light source making it difficult to recognise the features of the Fata Morgana that are obvious in daylight and with its unsuspected great distance contributing to the mystery of its origins. CONCLUSION: Many of the strange properties of the Min Min light are explicable in terms of the unusual optical conditions of the Fata Morgana, if account is also taken of the human factors that operate under these highly-reduced stimulus conditions involving a single isolated light source without reference landmarks. PMID- 12643808 TI - Acute macular neuroretinopathy. AB - Acute macular neuroretinopathy is an infrequently encountered condition in which there is a sudden mild central vision loss, photopsia and red-brown wedge-shaped lesions in the macular region with corresponding scotomata in the central visual fields. The condition may be associated with the use of oral contraceptives or with a recent febrile illness. It is self-limiting and non-recurrent. The clinical features of a patient with acute macular neuroretinopathy are described and the nature of the vascular aetiology is discussed. PMID- 12643809 TI - Leon F Garner. Foundation professor of optometry at the University of Auckland 1989-2003. PMID- 12643811 TI - Central venous catheter related infections: risk factors and the effect of glycopeptide antibiotics. AB - BACKGROUND: We undertook a prospective study of all new central venous catheters inserted into patients in the intensive care units, in order to identify the risk factors and to determine the effect of glycopeptide antibiotics on catheter - related infections. METHODS: During the study period 300 patients with central venous catheters were prospectively studied. The catheters used were nontunneled, noncuffed, triple lumen and made of polyurethane material. Catheters were cultured by semiquantitative method and blood cultures done when indicated. Data were obtained on patient age, gender, unit, primary diagnosis on admission, catheter insertion site, duration of catheterization, whether it was the first or a subsequent catheter and glycopeptide antibiotic usage. RESULTS: Ninety-one (30.3%) of the catheters were colonized and infection was found with 50 (16.7%) catheters. Infection was diagnosed with higher rate in catheters inserted via jugular vein in comparison with subclavian vein (95% CI: 1.32-4.81, p = 0.005). The incidence of infection was higher in catheters which were kept in place for more than seven days (95% CI 1.05-3.87, p = 0.03). The incidence of infection was lower in patients who were using glycopeptide antibiotic during catheterization (95% CI: 1.49-5.51, p = 0.005). The rate of infection with Gram positive cocci was significantly lower in glycopeptide antibiotic using patients (p = 0.01). The most commonly isolated organism was Staphylococcus aureus (n = 52, 37.1%). CONCLUSION: Duration of catheterization and catheter insertion site were independent risk factors for catheter related infection. Use of glycopeptide antibiotic during catheterization seems to have protective effect against catheter related infection. PMID- 12643810 TI - Variation in the pH-dependent pre-steady-state and steady-state kinetic characteristics of cysteine-proteinase mechanism: evidence for electrostatic modulation of catalytic-site function by the neighbouring carboxylate anion. AB - The acylation and deacylation stages of the hydrolysis of N -acetyl-Phe-Gly methyl thionoester catalysed by papain and actinidin were investigated by stopped flow spectral analysis. Differences in the forms of pH-dependence of the steady state and pre-steady-state kinetic parameters support the hypothesis that, whereas for papain, in accord with the traditional view, the rate-determining step is the base-catalysed reaction of the acyl-enzyme intermediate with water, for actinidin it is a post-acylation conformational change required to permit release of the alcohol product and its replacement in the catalytic site by the key water molecule. Possible assignments of the kinetically influential p K (a) values, guided by the results of modelling, including electrostatic-potential calculations, and of the mechanistic roles of the ionizing groups, are discussed. It is concluded that Asp(161) is the source of a key electrostatic modulator (p K (a) 5.0+/-0.1) in actinidin, analogous to Asp(158) in papain, whose influence is not detected kinetically; it is always in the 'on' state because of its low p K (a) value (2.8+/-0.06). PMID- 12643813 TI - Coping and caregivers of people with dementia. AB - PURPOSE: To critically review the research based on Lazarus and Folkman's (1984) stress and coping model, in respect to the coping of those caring for persons with dementia in the community, in an attempt to establish its implications for interventions aimed at improving caregiver adjustment. METHOD: Published material on the coping of caregivers of persons with dementia was identified through computerized literature searches (Med-line, Psych-Info) to December 1999, employing search terms including Alzheimer's disease, dementia, caregiving, caregiver burden, adaptation, psychological, coping, and stress. Studies were chosen to be considered in detail, based on the reviewer's opinion that they would contribute to an understanding of the current state of the research and its clinical implications. This material was then critically reviewed against the tenets of Lazarus and Folkman's (1984) model. RESULTS: Sixteen studies were selected to be included in the review, 12 cross-sectional and 4 longitudinal. Seven of the studies did not incorporate coping measures specific to caregiving and/or assess coping in respect of specific caregiver problems. Nine of the studies did do this. The research suggests that a general tendency towards problemsolving and acceptance styles of coping is likely to be advantageous to caregivers of people with dementia. CONCLUSIONS: Despite this finding, it is concluded that the ability of the research to inform the clinician is severely limited. It is proposed that while longitudinal studies considered specific caregiver problems which incorporate coping measures specific to the caregiving task may improve understanding, a substantial revision of methodology and perspective may be required to produce findings that are likely to influence practice. PMID- 12643812 TI - Epidemiological models for the spread of anti-malarial resistance. AB - BACKGROUND: The spread of drug resistance is making malaria control increasingly difficult. Mathematical models for the transmission dynamics of drug sensitive and resistant strains can be a useful tool to help to understand the factors that influence the spread of drug resistance, and they can therefore help in the design of rational strategies for the control of drug resistance. METHODS: We present an epidemiological framework to investigate the spread of anti-malarial resistance. Several mathematical models, based on the familiar Macdonald-Ross model of malaria transmission, enable us to examine the processes and parameters that are critical in determining the spread of resistance. RESULTS: In our simplest model, resistance does not spread if the fraction of infected individuals treated is less than a threshold value; if drug treatment exceeds this threshold, resistance will eventually become fixed in the population. The threshold value is determined only by the rates of infection and the infectious periods of resistant and sensitive parasites in untreated and treated hosts, whereas the intensity of transmission has no influence on the threshold value. In more complex models, where hosts can be infected by multiple parasite strains or where treatment varies spatially, resistance is generally not fixed, but rather some level of sensitivity is often maintained in the population. CONCLUSIONS: The models developed in this paper are a first step in understanding the epidemiology of anti-malarial resistance and evaluating strategies to reduce the spread of resistance. However, specific recommendations for the management of resistance need to wait until we have more data on the critical parameters underlying the spread of resistance: drug use, spatial variability of treatment and parasite migration among areas, and perhaps most importantly, cost of resistance. PMID- 12643814 TI - Relationships between personality, an extended theory of planned behaviour model and exercise behaviour. AB - OBJECTIVES: The purpose of this study was to investigate the theory of planned behaviour's (TPB) mediating hypothesis between the five-factor model of personality and exercise behaviour using an extended TPB model including concepts of affective and instrumental attitude, injunctive and descriptive norm, controllability, and selfefficacy. It was hypothesized that extraversion's activity facet would have a significant direct effect on exercise behaviour while controlling for the TPB, based on the presupposition that activity may represent a disposition that predicts exercise beyond planned behaviour. DESIGN: To test the replicability of these findings, we examined this research question with undergraduate students prospectively and cancer survivors, using a cross sectional design. RESULTS: Using structural equation modelling, the results indicated that activity had a significant effect (p <.05) on exercise behaviour (study 1 =.20; study 2 =.31) while controlling for the TPB. CONCLUSIONS: This study suggests the importance of extraversion's activity facet on exercise behaviour, even when controlling for a TPB model with additional socialcognitive concepts and disparate population samples. PMID- 12643815 TI - The expectancy-value muddle in the theory of planned behaviour - and some proposed solutions. AB - The authors of the Theories of Reasoned Action and Planned Behaviour recommended a method for statistically analysing the relationships between beliefs and the Attitude, Subjective Norm, and Perceived Behavioural Control constructs. This method has been used in the overwhelming majority of studies using these theories. However, there is a growing awareness that this method yields statistically uninterpretable results (Evans, 1991). Despite this, the use of this method is continuing, as is uninformed interpretation of this problematic research literature. This is probably due to the lack of a simple account of where the problem lies, and the large number of alternatives available. This paper therefore summarizes the problem as simply as possible, gives consideration to the conclusions that can be validly drawn from studies that contain this problem, and critically reviews the many alternatives that have been proposed to address this problem. Different techniques are identified as being suitable, according to the purpose of the specific research project. PMID- 12643816 TI - Nutritional supplementation in older adults: pleasantness, preference and selection of sip-feeds. AB - OBJECTIVES: Malnourishment is common in older adults, and nutritional supplementation is used to improve body weight and well-being. Clinical reports suggest, however, that patients routinely reject sip-feeds. The present study examined the following questions: whether sip-feeds are less preferred and less likely to be selected than other energy-dense foods in healthy elders; and whether eating alone further reduces intake relative to eating in a social setting. METHODS: Twenty-one healthy older adults (aged 60-79 years) attended the laboratory on three occasions. Subjects rated six different flavours of sip-feed and then rated the pleasantness of the taste of the favoured flavour against five other energy-dense familiar foods/drinks. Intake of these foods was measured when subjects ate alone or in a group of familiar others. RESULTS: Favourite flavour of sip-feed compared well with other more familiar foods and was selected as part of a snack. Snack intake increased by 60% when consumed in a group setting compared with eating alone. CONCLUSIONS: The study suggests that sip-feeds are rated as pleasant and selected by free-living elders. Rejection of sip-feeds in hospitalized elders may relate more to loss of appetite than to the taste preference for sip-feeds, and that eating alone rather than in groups of familiar others is likely to compound eating problems. PMID- 12643817 TI - Predicting adherence to eye patching in children with amblyopia: an application of protection motivation theory. AB - OBJECTIVES: This study reports an application of protection motivation theory (PMT) to the prediction of parental adherence to eye patching recommendations for children with amblyopia over a 2-month period. The study also considered the role of past behaviour in PMT. DESIGN AND METHODS: A total of 151 parents of children with amblyopia who were attending follow-up appointments for orthoptic treatment participated. They completed questionnaires based on PMT to assess their beliefs about amblyopia and eye patching. Of the parents, 105 were contacted again at 2 month follow-up to obtain a measure of adherence to the recommended treatment for their child. RESULTS: PMT was found to be predictive of adherence intentions and behaviour at 2-month follow-up. Regression analyses revealed perceived vulnerability, response efficacy and self-efficacy to be significant predictors of protection motivation, whereas perceived vulnerability and response costs were significant predictors of adherence behaviour. Past adherence behaviour was found to have a direct effect on future adherence behaviour over and above the influence of PMT. CONCLUSIONS: The results are discussed in relation to the sufficiency of PMTas a model of adherence behaviour. The practical implications for attempts to increase adherence to eye patching among children with amblyopia are outlined. PMID- 12643818 TI - A contemporary validation of the Reeder Stress Inventory. AB - OBJECTIVES: To examine the construct validity of the RSI in a contemporary cohort. DESIGN: A cross-sectional investigation of 1,717 employed individuals who responded to the second stage of a study of occupational stress. METHODS: Scores on the RSI are compared to smoking and drinking habits, social class, and two measures of health (number of days sick leave, and number of visits to a GP during the previous year). The RSI was compared to three questionnaires measuring concepts related to stress: the 12-item General Health Questionnaire (GHQ-12), the Hospital Anxiety and Depression Scale (HADS), and the Karasek Job Strain Questionnaire. RESULTS: Higher levels of stress, as measured by the RSI, were associated with smoking a greater number of cigarettes, and, if the respondent drank alcohol, greater consumption of alcoholic drinks. High levels of stress were also associated with having taken more days sick leave, having made more frequent visits to a GP, and, somewhat unexpectedly, with being a teetotaller, and with holding a non-manual occupation. Of the questionnaire measures, the strongest association was between the RSI and HADS anxiety subscale, consistent with an overlap between the concepts of stress and anxiety. CONCLUSIONS: This study supports the construct validity of the RSI in a sample of employed individuals. PMID- 12643819 TI - How sleep is related to fatigue. AB - OBJECTIVES: It is evident that sleep patterns have direct effects on fatigue. However, the multidimensionality of fatigue may imply that complex patterns of relationships exist between fatigue and sleep characteristics. We aimed to study the correlations between fatigue and quantitative and qualitative sleep measurements, while taking into consideration depression and somatization which are known to affect both sleep and fatigue. We predicted that sleep quality, unattained by the effects of somatization and depression, would affect perceived fatigue more than the quantitative characteristics of sleep. DESIGN: Employing a cross-sectional design, hypotheses were addressed using multiple hierarchical regression analyses according to established methods. METHODS: Data were gathered from a targeted, randomly selected adult sample (N = 278) by means of subjective sleep reports, a mental health inventory, somatization inventory, several fatigue questionnaires and a demographic questionnaire. RESULTS: Fatigue was significantly predicted by depression scores, somatization levels and subjective sleep quality, but not quantitative sleep characteristics such as sleep latency, nocturnal awakenings and early morning arousals. Depression levels were positively and significantly related to all aspects of fatigue except physical fatigue and fatigue that responds to rest and sleep. Physical fatigue was correlated with somatization, but not depression. CONCLUSIONS: The data further our understanding of the multifaceted nature of human fatigue and underline the greater importance of perceived sleep quality, compared to other sleep characteristics, in predicting fatigue. PMID- 12643820 TI - Evidence for the independence of positive and negative well-being: implications for quality of life assessment. AB - OBJECTIVES: Evidence is accumulating that positive mental states are more than the absence of symptoms, and may play an independent role in health outcomes. The aim of this study is to compare the characteristics and determinants of positive and negative mental states in a population sample. DESIGN: A novel analysis of data was undertaken from the General Health Questionnaire (GHQ-30) which was completed by 6,317 participants in the Health and Lifestyle Survey at Time 1 and 3,778 at Time 2, 7 years later. METHODS: We derived a positive well-being scale (POS-GHQ) based on positive responses to the positive items of the GHQ-30, and compared it to a standard symptom measure (CGHQ). Discriminant function analyses were performed to establish which demographic, health and social variables best accounted for scores on each scale. RESULTS: The distributional properties of the two scales, together with the results of the discriminant analyses, demonstrate a degree of independence between positive and negative well-being. Over one third of the sample obtained either low scores on both positive and negative well-being measures or high scores on both measures. Disability and lack of social roles were important determinants of psychological symptoms, but had less influence on positive well-being. Having paid employment was an important determinant of positive well-being but had less influence on psychological symptoms. We also found that 7-year mortality was predicted more strongly by the absence of positive well-being than by the presence of psychological symptoms. CONCLUSIONS: These findings point to the need to include measures of positive well-being in studies of health outcomes and quality of life assessment. PMID- 12643821 TI - Emerging trends in international law concerning global infectious disease control. AB - International cooperation has become critical in controlling infectious diseases. In this article, I examine emerging trends in international law concerning global infectious disease control. The role of international law in horizontal and vertical governance responses to infectious disease control is conceptualized; the historical development of international law regarding infectious diseases is described; and important shifts in how states, international institutions, and nonstate organizations use international law in the context of infectious disease control today are analyzed. The growing importance of international trade law and the development of global governance mechanisms, most prominently in connection with increasing access to drugs and other medicines in unindustrialized countries, are emphasized. Traditional international legal approaches to infectious disease control--embodied in the International Health Regulations--may be moribund. PMID- 12643822 TI - Enterovirus 71 outbreaks, Taiwan: occurrence and recognition. AB - Enterovirus 71 (EV71) caused a large outbreak in Taiwan in 1998 with 78 deaths, and smaller outbreaks recurred in 2000 and 2001. The outbreak was recognized because of a large number of hand, foot, and mouth disease cases and the rapid deaths of children with the disease. Virologic and pathologic studies indicated that EV71 was the most important agent related to severe and fatal cases and that a neurogenic inflammatory response was involved in the pathogenesis of cardiopulmonary collapse resulting from fulminant EV71 infection. Seroepidemiologic study suggested that EV71 had circulated for at least 16 years and that the accumulation of susceptible hosts might have triggered the 1998 outbreak. However, a change in EV71 neurovirulence and host genetic susceptibility may also have affected the clinical outcome. The Taiwan outbreak shows that worldwide attention should be paid to such outbreaks, new antiviral drugs should be developed, and that vaccination of children under 5 years of age may be warranted. PMID- 12643823 TI - Electron microscopy for rapid diagnosis of infectious agents in emergent situations. AB - Diagnostic electron microscopy has two advantages over enzyme-linked immunosorbent assay and nucleic acid amplification tests. After a simple and fast negative stain preparation, the undirected, "open view" of electron microscopy allows rapid morphologic identification and differential diagnosis of different agents contained in the specimen. Details for efficient sample collection, preparation, and particle enrichment are given. Applications of diagnostic electron microscopy in clinically or epidemiologically critical situations as well as in bioterrorist events are discussed. Electron microscopy can be applied to many body samples and can also hasten routine cell culture diagnosis. To exploit the potential of diagnostic electron microscopy fully, it should be quality controlled, applied as a frontline method, and be coordinated and run in parallel with other diagnostic techniques. PMID- 12643824 TI - Influenza AH1N2 viruses, United Kingdom, 2001-02 influenza season. AB - During the winter of 2001-02, influenza AH1N2 viruses were detected for the first time in humans in the U.K. The H1N2 viruses co-circulated with H3N2 viruses and a very small number of H1N1 viruses and were isolated in the community and hospitalized patients, predominantly from children <15 years of age. Characterization of H1N2 viruses indicated that they were antigenically and genetically homogeneous, deriving the hemagglutinin (HA) gene from recently circulating A/New Caledonia/20/99-like H1N1 viruses, whereas the other seven genes originated from recently circulating H3N2 viruses. Retrospective reverse transcription-polymerase chain reaction analysis of influenza A H1 viruses isolated in the U.K. during the previous winter identified a single H1N2 virus, isolated in March 2001, indicating that H1N2 viruses did not widely circulate in the U.K. before September 2001. The reassortment event is estimated to have occurred between 1999 and early 2001, and the emergence of H1N2 viruses in humans reinforces the need for frequent surveillance of circulating viruses. PMID- 12643825 TI - Experimental infection of North American birds with the New York 1999 strain of West Nile virus. AB - To evaluate transmission dynamics, we exposed 25 bird species to West Nile virus (WNV) by infectious mosquito bite. We monitored viremia titers, clinical outcome, WNV shedding (cloacal and oral), seroconversion, virus persistence in organs, and susceptibility to oral and contact transmission. Passeriform and charadriiform birds were more reservoir competent (a derivation of viremia data) than other species tested. The five most competent species were passerines: Blue Jay (Cyanocitta cristata), Common Grackle (Quiscalus quiscula), House Finch (Carpodacus mexicanus), American Crow (Corvus brachyrhynchos), and House Sparrow (Passer domesticus). Death occurred in eight species. Cloacal shedding of WNV was observed in 17 of 24 species, and oral shedding in 12 of 14 species. We observed contact transmission among four species and oral in five species. Persistent WNV infections were found in tissues of 16 surviving birds. Our observations shed light on transmission ecology of WNV and will benefit surveillance and control programs. PMID- 12643826 TI - Emergence of ceftriaxone-resistant Salmonella isolates and rapid spread of plasmid-encoded CMY-2-like cephalosporinase, Taiwan. AB - Of 384 Salmonella isolates collected from 1997 to 2000 in a university hospital in Taiwan, six ceftriaxone-resistant isolates of Salmonella enterica serovar Typhimurium were found in two patients in 2000. The resistance determinants were on conjugative plasmids that encoded a CMY-2-like cephalosporinase. During the study period, the proportion of CMY-2-like enzyme producers among Escherichia coli increased rapidly from 0.2% in early 1999 to >4.0% in late 2000. Klebsiella pneumoniae isolates producing a CMY-2-like beta-lactamase did not emerge until 2000. The presence of bla(CMY)-containing plasmids with an identical restriction pattern from Salmonella, E. coli, and K. pneumoniae isolates was found, which suggests interspecies spread and horizontal transfer of the resistance determinant. Various nosocomial and community-acquired infections were associated with the CMY-2-like enzyme producers. Our study suggests that the spread of plasmid-mediated CMY-2-like beta-lactamases is an emerging threat to hospitalized patients and the public in Taiwan. PMID- 12643827 TI - Bartonella henselae in Ixodes ricinus ticks (Acari: Ixodida) removed from humans, Belluno province, Italy. AB - The potential role of ticks as vectors of Bartonella species has recently been suggested. In this study, we investigated the presence of Bartonella species in 271 ticks removed from humans in Belluno Province, Italy. By using primers derived from the 60-kDa heat shock protein gene sequences, Bartonella DNA was amplified and sequenced from four Ixodes ricinus ticks (1.48%). To confirm this finding, we performed amplification and partial sequencing of the pap31 protein and the cell division protein ftsZ encoding genes. This process allowed us to definitively identify B. henselae (genotype Houston-1) DNA in the four ticks. Detection of B. henselae in these ticks might represent a highly sensitive form of xenodiagnosis. B. henselae is the first human-infecting Bartonella identified from Ixodes ricinus, a common European tick and the vector of various tickborne pathogens. The role of ticks in the transmission of bartonellosis should be further investigated. PMID- 12643828 TI - New lyssavirus genotype from the Lesser Mouse-eared Bat (Myotis blythi), Kyrghyzstan. AB - The Aravan virus was isolated from a Lesser Mouse-eared Bat (Myotis blythi) in the Osh region of Kyrghyzstan, central Asia, in 1991. We determined the complete sequence of the nucleoprotein (N) gene and compared it with those of 26 representative lyssaviruses obtained from databases. The Aravan virus was distinguished from seven distinct genotypes on the basis of nucleotide and amino acid identity. Phylogenetic analysis based on both nucleotide and amino acid sequences showed that the Aravan virus was more closely related to genotypes 4, 5, and--to a lesser extent--6, which circulates among insectivorus bats in Europe and Africa. The Aravan virus does not belong to any of the seven known genotypes of lyssaviruses, namely, rabies, Lagos bat, Mokola, and Duvenhage viruses and European bat lyssavirus 1, European bat lyssavirus 2, and Australian bat lyssavirus. Based on these data, we propose a new genotype for the Lyssavirus genus. PMID- 12643829 TI - Molecular detection of Bartonella quintana, B. koehlerae, B. henselae, B. clarridgeiae, Rickettsia felis, and Wolbachia pipientis in cat fleas, France. AB - The prevalences of Bartonella, Rickettsia, and Wolbachia were investigated in 309 cat fleas from France by polymerase chain reaction (PCR) assay and sequencing with primers derived from the gltA gene for Rickettsia, the its and pap31 genes for Bartonella, and the 16S rRNA gene for Anaplasmataceae. Positive PCR results were confirmed by using the Lightcycler and specific primers for the rOmpB of Rickettsia and gltA of Bartonella. R. felis was detected in 25 fleas (8.1%), W. pipientis, an insect symbiont, in 55 (17.8%), and Bartonella in 81 (26.2%), including B. henselae (9/81; 11.1%), B. clarridgeiae (55/81; 67.9%), B. quintana (14/81; 17.3%), and B. koehlerae (3/81; 3.7%). This is the first report of the amplification of B. quintana from fleas and the first description of B. koehlerae in fleas from an area outside the United States. Cat fleas may be more important vectors of human diseases than previously reported. PMID- 12643830 TI - European echinococcosis registry: human alveolar echinococcosis, Europe, 1982 2000. AB - Surveillance for alveolar echinococcosis in central Europe was initiated in 1998. On a voluntary basis, 559 patients were reported to the registry. Most cases originated from rural communities in regions from eastern France to western Austria; single cases were reported far away from the disease-"endemic" zone throughout central Europe. Of 210 patients, 61.4% were involved in vocational or part-time farming, gardening, forestry, or hunting. Patients were diagnosed at a mean age of 52.5 years; 78% had symptoms. Alveolar echinococcosis primarily manifested as a liver disease. Of the 559 patients, 190 (34%) were already affected by spread of the parasitic larval tissue. Of 408 (73%) patients alive in 2000, 4.9% were cured. The increasing prevalence of Echinococcus multilocularis in foxes in rural and urban areas of central Europe and the occurrence of cases outside the alveolar echinococcosis-endemic regions suggest that this disease deserves increased attention. PMID- 12643831 TI - Tularemia on Martha's Vineyard: seroprevalence and occupational risk. AB - We conducted a serosurvey of landscapers to determine if they were at increased risk for exposure to Francisella tularensis and to determine risk factors for infection. In Martha's Vineyard, Massachusetts, landscapers (n=132) were tested for anti-F. tularensis antibody and completed a questionnaire. For comparison, serum samples from three groups of nonlandscaper Martha's Vineyard residents (n=103, 99, and 108) were tested. Twelve landscapers (9.1%) were seropositive, compared with one person total from the comparison groups (prevalence ratio 9.0; 95% confidence interval 1.2 to 68.1; p=0.02). Of landscapers who used a power blower, 15% were seropositive, compared to 2% who did not use a power blower (prevalence ratio 9.2; 95% confidence interval 1.2 to 69.0; p=0.02). Seropositive landscapers worked more hours per week mowing and weed-whacking and mowed more lawns per week than their seronegative counterparts. Health-care workers in tularemia-endemic areas should consider tularemia as a diagnosis for landscapers with a febrile illness. PMID- 12643832 TI - Epidemiology of meningococcal disease, New York City, 1989-2000. AB - Study of the epidemiologic trends in meningococcal disease is important in understanding infection dynamics and developing timely and appropriate public health interventions. We studied surveillance data from the New York City Department of Health and Mental Hygiene, which showed that during 1989-2000 a decrease occurred in both the proportion of patients with serogroup B infection (from 28% to 13% of reported cases; p<0.01) and the rate of serogroup B infection (from 0.25/100,000 to 0.08/100,000; p<0.01). We also noted an increased proportion (from 3% to 39%; p<0.01) and rate of serogroup Y infection (from 0.02/100,000 to 0.23/100,000; p<0.01). Median patient age increased (from 15 to 30 years; p<0.01). The case-fatality rate for the period was 17%. As more effective meningococcal vaccines become available, recommendations for their use in nonepidemic settings should consider current epidemiologic trends, particularly changes in age and serogroup distribution of meningococcal infections. PMID- 12643833 TI - Amplification of the sylvatic cycle of dengue virus type 2, Senegal, 1999-2000: entomologic findings and epidemiologic considerations. AB - After 8 years of silence, dengue virus serotype 2 (DENV-2) reemerged in southeastern Senegal in 1999. Sixty-four DENV-2 strains were isolated in 1999 and 9 strains in 2000 from mosquitoes captured in the forest gallery and surrounding villages. Isolates were obtained from previously described vectors, Aedes furcifer, Ae. taylori, Ae. luteocephalus, and--for the first time in Senegal- from Ae. aegypti and Ae. vittatus. A retrospective analysis of sylvatic DENV-2 outbreaks in Senegal during the last 28 years of entomologic investigations shows that amplifications are periodic, with intervening, silent intervals of 5-8 years. No correlation was found between sylvatic DENV-2 emergence and rainfall amount. For sylvatic DENV-2 vectors, rainfall seems to particularly affect virus amplification that occurs at the end of the rainy season, from October to November. Data obtained from investigation of preimaginal (i.e., nonadult) mosquitoes suggest a secondary transmission cycle involving mosquitoes other than those identified previously as vectors. PMID- 12643834 TI - Rabies in Sri Lanka: splendid isolation. AB - Rabies virus exists in dogs on Sri Lanka as a single, minimally divergent lineage only distantly related to other rabies virus lineages in Asia. Stable, geographically isolated virus populations are susceptible to local extinction. A fully implemented rabies-control campaign could make Sri Lanka the first Asian country in >30 years to become free of rabies virus. PMID- 12643835 TI - Human metapneumovirus in severe respiratory syncytial virus bronchiolitis. AB - Reverse transcription-polymerase chain reaction was used to detect segments of the M (matrix), N (nucleoprotein), and F (fusion) genes of human metapneumovirus in bronchoalveolar fluid from 30 infants with severe respiratory syncytial virus bronchiolitis. Seventy percent of them were coinfected with metapneumovirus. Such coinfection might be a factor influencing the severity of bronchiolitis. PMID- 12643836 TI - Persistence of virus-reactive serum immunoglobulin m antibody in confirmed west nile virus encephalitis cases. AB - Twenty-nine laboratory-confirmed West Nile virus (WNV encephalitis patients were bled serially so that WNV-reactive immunoglobulin (Ig) M activity could be determined. Of those patients bled, 7 (60%) of 12 had anti-WNV IgM at approximately 500 days after onset. Clinicians should be cautious when interpreting serologic results from early season WNV IgM-positive patients. PMID- 12643838 TI - Echinococcus multilocularis: an emerging pathogen in Hungary and Central Eastern Europe? AB - Echinococcus multilocularis, the causative agent of human alveolar echinococcosis, is reported for the first time in Red Foxes (Vulpes vulpes) in Hungary. This parasite may be spreading eastward because the population of foxes has increased as a consequence of human interventions, and this spread may result in the emergence of alveolar echinococcosis in Central Eastern Europe. PMID- 12643837 TI - Isolation of Escherichia coli O157:H7 from intact colon fecal samples of swine. AB - Escherichia coli O157:H7 was recovered from colon fecal samples of pigs. Polymerase chain reaction confirmed two genotypes: isolates harboring the eaeA, stx(1), and stx(2) genes and isolates harboring the eaeA, stx(1), and hly(933) genes. We demonstrate that swine in the United States can harbor potentially pathogenic E. coli O157:H7. PMID- 12643839 TI - Early and definitive diagnosis of toxic shock syndrome by detection of marked expansion of T-cell-receptor VBeta2-positive T cells. AB - We describe two cases of early toxic shock syndrome, caused by the superantigen produced from methicillin-resistant Staphylococcus aureus and diagnosed on the basis of an expansion of T-cell-receptor VBeta2-positive T cells. One case patient showed atypical symptoms. Our results indicate that diagnostic systems incorporating laboratory techniques are essential for rapid, definitive diagnosis of toxic shock syndrome. PMID- 12643841 TI - The National Capitol Region's Emergency Department syndromic surveillance system: do chief complaint and discharge diagnosis yield different results? AB - We compared syndromic categorization of chief complaint and discharge diagnosis for 3,919 emergency department visits to two hospitals in the U.S. National Capitol Region. Agreement between chief complaint and discharge diagnosis was good overall (kappa=0.639), but neurologic and sepsis syndromes had markedly lower agreement than other syndromes (kappa statistics 0.085 and 0.105, respectively). PMID- 12643840 TI - Removing deer mice from buildings and the risk for human exposure to Sin Nombre virus. AB - Trapping and removing deer mice from ranch buildings resulted in an increased number of mice, including Sin Nombre virus antibody-positive mice, entering ranch buildings. Mouse removal without mouse proofing will not reduce and may even increase human exposure to Sin Nombre hantavirus. PMID- 12643842 TI - Asymptomatic visceral leishmaniasis, northern Israel. AB - Asymptomatic human visceral leishmaniasis was identified in Israel by using an enzyme-linked immunosorbent assay. Positive serum samples were more prevalent in visceral leishmaniasis-endemic (2.97%) compared to nonendemic (1.01%) regions (p=0.021). Parasite exposure was higher than expected, despite the small number of clinical cases, suggesting factors other than infection per se influence clinical outcome. PMID- 12643844 TI - Emerging human infectious diseases: anthroponoses, zoonoses, and sapronoses. PMID- 12643843 TI - Mycobacterium celatum pulmonary infection in the immunocompetent: case report and review. AB - Mycobacterium celatum has been shown to cause disease in immunocompromised patients. We report a case of serious pulmonary infection caused by M. celatum in an apparently immunocompetent patient and review the characteristics of two other reported cases. Clinical and radiologic symptoms and signs included cough, malaise, and weight loss associated with cavitary lesions and pulmonary infiltrates. Although M. celatum is easy to detect in clinical specimens by liquid and solid media, it may be misidentified as a member of the M. tuberculosis complex or as M. xenopi. M. celatum pulmonary infection appears to respond to antimycobacterial chemotherapy, particularly with clarithromycin. PMID- 12643845 TI - Multidrug-resistant Shigella dysenteriae type 1: forerunners of a new epidemic strain in eastern India? PMID- 12643847 TI - Changes in epithelial cell turnover and extracellular matrix in human small intestine after TPN. AB - BACKGROUND: The atrophy and architectural remodeling of the jejunal mucosa arising in adults receiving total parenteral nutrition (TPN) has been suggested to originate from a disturbance in tissue homeostasis. The present study aims at examining (1) whether there are differences in proliferation and apoptosis of epithelial cells between enterally and parenterally nourished patients and (2) whether the distribution pattern of extracellular matrix (ECM) proteins known to influence cell turnover along the the crypt-villus axis is changed after TPN. METHODS: The mitotic frequency and the proliferation index [using an antibody against Ki-67 antigen (MIB 1)] were determined on epoxy semithin and paraffin sections, respectively. Morphological techniques and the TUNEL assay were applied to detect apoptotic events. Immunolocalization of collagen IV, laminin, fibronectin, tenascin, and collagen VI was performed on cryosections. RESULTS: After TPN the cell renewal was significantly enhanced, while epithelial cell death was drastically reduced. The comparison of TPN and EN patients revealed differences in the distribution patterns of the ECM proteins laminin, fibronectin, and tenascin along the crypt-villus axis. Moreover, after TPN an increased expression of collagen types IV and VI was observed. CONCLUSIONS: TPN in human adults is associated with alterations in epithelial cell turnover and changes in expression and/or localization of ECM proteins. Thus, the inverted route of nutrient supply in patients might modify environmental tissue conditions, which may influence the interactions between intestinal epithelial cells and the extracellular matrix. PMID- 12643846 TI - K(ATP) channel opener protects neonatal rabbit heart better than St. Thomas' solution. AB - OBJECTIVES: Myocardial protection with ATP-sensitive potassium channel (K(ATP) channel) openers is as effective as St. Thomas' cardioplegia (StTCP) in adult rabbit hearts. This study compares the effectiveness of the K(ATP) channel opener pinacidil to StTCP in protecting neonatal rabbit hearts exposed to global ischemia. METHODS: Seventeen neonatal rabbit hearts (7-9 days old) perfused with Krebs-Henseleit buffer (KHB) on a Langendorff apparatus underwent 90 min of normothermic ischemia. Six (ischemia control) received no pretreatment before or during ischemia. Six others (pinacidil) received a 3-min infusion of 50 microM pinacidil in KHB without StTCP at the onset of ischemia. Five others (StTCP) received a 3-min infusion of StTCP at the onset of ischemia. After 60 min of KHB reperfusion, recovery of left ventricular (LV) performance and coronary flow (CF) were measured and compared to preischemia. A paired t test was used for comparison between drug-treated and untreated groups. RESULTS: Pinacidil-treated hearts had significantly better recovery of left ventricular developed pressure (47 +/- 3.8 mmHg vs 32 +/- 2.5 mmHg, P < 0.05), contractility (+dP/dt(max); 885.4 +/- 74 mmHg vs 643.7 +/- 65 mmHg, P < 0.05), left ventricular end diastolic pressure (10.5 +/- 0.9 mmHg vs 17.4 +/- 1.2 mmHg P < 0.05), compliance ( dP/dt(max); 994.2 +/- 86 mmHg vs 673.6 +/- 69 mmHg, P < 0.05), and CF (5.9 +/- 0.4 ml/min vs 4.2 +/- 0.2 ml/min, P < 0.05) compared to ischemic control. StTCP only improved the recovery of -dP/dt(max) (877.4 +/- 73 mmHg/s vs 673.6 +/- 69 mmHg/s, P < 0.05) and CF (5.7 +/- 0.3 ml/min vs 4.2 +/- 0.2 ml/min, P < 0.05) compared to control. CONCLUSIONS: Pinacidil pretreatment provided superior recovery of systolic performance compared to St. Thomas' cardioplegia solution in neonatal hearts. Myocardial protection by pretreatment with the K(ATP) channel opener pinacidil may be a new strategy for myocardial protection during pediatric cardiac surgery. PMID- 12643848 TI - Clinical significance of tissue inhibitor of metalloproteinase and matrix metalloproteinase mRNA expression in thymoma. AB - BACKGROUND: Matrix metalloproteinases (MMPs) are proteolytic enzymes which degrade extracellular matrix and basement membrane. There is much evidence that their increased expression is correlated with tumor aggressiveness in various carcinomas. Tissue inhibitor of metalloproteinases (TIMPs) are the specific inhibitors of MMPs. MMPs and TIMPs are considered to play an important role in carcinoma invasion and metastasis. We hypothesized that MMPs and TIMPs also play an important role in thymoma. MATERIALS AND METHODS: This study included 34 thymoma cases. The mRNA levels of MMP-1, -7, and -9, TIMP-1 and -2, and GAPDH were quantified by real-time polymerase chain reaction using LightCycler. We also performed immunohistochemistry for TIMP-1. RESULTS: The TIMP-1/GAPDH mRNA expression level was significantly higher in invasive (stage II-IV) thymomas (means +/- SE, 81.4 +/- 28.1) than in noninvasive (stage I) thymomas (30.9 +/- 8.3, P = 0.026). The MMP-1/GAPDH mRNA expression level was also higher in invasive thymomas (19.7 +/- 7.5) than in non invasive thymomas (2.26 +/- 1.72, P = 0.020). Immunopositivity of TIMP-1 was localized in stromal cells adjacent to the advancing margin of the tumor. CONCLUSIONS: These findings suggest that TIMPs and MMPs play an important role in the invasion of thymoma. PMID- 12643849 TI - Influence of insulin therapy on burn wound healing in rats. AB - BACKGROUND: Insulin is proposed as a therapy for suppressing muscle wasting after burn trauma although the long-term effects of this therapy on wound healing are not yet known. The present study was designed to investigate the effect of systemically administered insulin therapy on burn wound healing. MATERIALS AND METHODS: Young rats weighing 80-150 g were subjected to 15-20% total body surface area burn injury on their shaved dorsum. The insulin dosage was increased over the first 3 days in each rat from 0.25 U (Day 1), 0.5 U (Day 2), and 1.0 U (Day 3) per 100 g body wt. The rats were euthanized at the fourth or fifteenth day postinjury. Skin sections were analyzed by histochemistry and quantitative polarization microscopy. RESULTS: Histology showed a decreased number of inflammatory cells and increased vasodilation in the insulin-treated animals at Day 4 relative to untreated rats; at Day 15 there was increased reepithelialization. Quantitative analysis using polarization microscopy and picrosirius red staining showed an increased collagen deposition in wounds by Day 4 in insulin-treated rats relative to untreated burn controls. CONCLUSION: These results indicate that insulin induces accelerated wound healing associated with diminished inflammation and increased collagen deposition. PMID- 12643850 TI - The role of nitric oxide after a short period of liver ischemia-reperfusion. AB - BACKGROUND: Liver ischemia-reperfusion injury is a serious problem during liver resection and transplantation. Nitric oxide (NO) has been suggested to have a cytoprotective effect for microcirculation, while the interaction of active oxygen species and NO produces peroxynitrite anion. The present study attempts to clarify the role of NO in liver ischemia-reperfusion injury. METHODS: Wistar male rats were subjected to 30 min of hepatic ischemia followed by reperfusion. The model rats were divided into the three following groups: a control group that was not administered NO synthase inhibitors, and two experimental groups that were administered either N(omega)-nitro-L-arginine methyl ester (L-NAME) or aminoguanidine. In each group, we examined active oxygen species and nitric oxide production, and investigated liver function by measuring serum transaminase levels. In addition, we conducted histopathologic examinations and microcirculation examinations using intravital videomicroscopy. RESULTS: In the control group, NO concentrations in the plasma increased with time after reperfusion. A decrease in NO production was detected in the groups administered NO synthase inhibitors. Elevated serum transaminase levels became more prominent after L-NAME administration, while aminoguanidine administration reduced its level. The degree of microcirculation failure was found to be more prominent in the L-NAME-administered group over both the control group and the aminoguanidine administered group. A significantly lower survival rate was observed at 6 h after reperfusion in the L-NAME-administered group over that of the other groups. CONCLUSIONS: A reduction of the ischemia-reperfusion injury is important in inhibiting the production of high-output NO and peroxynitrite, and in maintaining NO levels necessary for maintenance of microcirculation. PMID- 12643851 TI - Inhibition of matrix metalloproteinases reduces local and distant organ injury following experimental acute pancreatitis. AB - BACKGROUND: Pulmonary complications from pancreatitis involve parenchymal destruction via proteolytic enzymes. Matrix metalloproteinases (MMPs) may play an important role in pulmonary injury following acute severe pancreatitis. We hypothesized that local and distant organ injury would be decreased by the presence of an MMP inhibitor (Batimistat; BB-94) following severe acute pancreatitis (AP). METHODS: Eighteen male rats were randomized into two groups: BB-94 (AP + 40 mg/kg/24 h BB-94 ip x three doses) or control (AP + 20 ml/kg/24 h normal saline ip x three doses). Necrotizing AP was induced by retrograde infusion of 5% sodium taurocholate (1.5 ml/kg) into the pancreatic duct. Twenty additional animals were randomized into BB-94 and control groups for the survival study. Serum was evaluated for amylase and MMP activity. Pancreatic sections were graded for edema, necrosis, neutrophil infiltrate, and hemorrhage. Myloperoxidase (MPO) activity was used to determine PMN infiltration in the lung. Evan's Blue dye extravasation was used to quantify vascular permeability. RESULTS: Animals in the BB-94 group had decreased amylase levels (1086.0 +/- 61.7 U/L vs 2232.7 +/- 309.9 U/L; P < 0.05), decreased cellular infiltrate (1.4 +/- 0.2 vs 2.3 +/- 0.2; P < 0.02), and decreased necrosis (4.1 +/- 0.3 vs 6.1 +/- 0.4; P < 0.005) compared to the control group. Lung tissue following pancreatitis in the BB-94 group demonstrated decreased MPO activity (41.5 +/- 2.4 units vs 57.3 +/- 2.9 units; P < 0.05) and decreased vascular permeability (18.3 +/- 2.8 mg/100 g vs 30.1 +/- 4.6 mg/100 g; P < 0.05). Animals treated with BB-94 had 100% survival compared to 50% survival in control at 72 h. CONCLUSIONS: Pancreatitis results in increased local and distant MMP activity. Pulmonary and pancreatic injury following AP can be abrogated by treatment with an MMP inhibitor (Batimistat; BB 94) which may result in decreased morbidity and mortality. PMID- 12643852 TI - Isolated lung perfusion with gemcitabine in a rat: pharmacokinetics and survival. AB - BACKGROUND: Toxicity and pharmacokinetics of gemcitabine (GCB) were evaluated in a rat model of isolated lung perfusion (ILuP) and compared to intravenous (iv) infusion. MATERIALS AND METHODS: CC531S adenocarcinoma cells were incubated in vitro for 24 h with GCB. Cell survival was determined 4 days after GCB treatment with the sulforhodamine B test. In a first in vivo experiment, Wag/Rij rats underwent left ILuP with 20 mg/kg (n = 3), 40 mg/kg (n = 6), 80 mg/kg (n = 6), 160 mg/kg (n = 6), or 320 mg/kg (n = 6) of GCB and a control group (n = 6) with buffered starch. After 3 weeks, right pneumonectomy was performed. Furthermore, survival was determined for rats treated with iv infusion of 40 mg/kg (n = 10), 80 mg/kg (n = 10), 160 mg/kg (n = 10), or 320 mg/kg (n = 6) of GCB and a control group (n = 6) treated with saline (0.9% NaCl). In a second experiment lung and serum GCB levels were determined for rats treated with iv infusion (160 mg/kg, n = 6) and rats which had ILuP (160 mg/kg, n = 6; 320 mg/kg, n = 6). RESULTS: Incubation of the CC531S adenocarcinoma cells with GCB led to a 50% decrease (P < 0.05) in the number of cells compared to controls at a dose of 23.6 nM. After 90 days, the mortality for rats treated with 320 mg/kg iv GCB was 100% compared to 17% after ILuP for the same dose. ILuP with 160 and 320 mg/kg resulted in significantly higher lung levels of GCB compared to iv therapy without any systemic leakage. CONCLUSIONS: GCB ILuP is well-tolerated to a maximum dose of 320 mg/kg and results in significantly higher GCB lung levels with undetectable serum levels compared to iv treatment. PMID- 12643853 TI - Epidermal growth factor stimulation can substitute for c-Src overexpression in promoting breast carcinoma invasion. AB - BACKGROUND AND AIMS: Cancer progression is in large part dependent on the complex process of cell invasion, involving adhesion, motility, and enzymatic proteolysis. Overexpression of the Src proto-oncogene (c-Src), a nonreceptor tyrosine kinase, has been implicated in the progression of both colon and breast cancer. Our group has previously reported that overexpression of c-Src leads to a significant gain in invasive cell behavior in vitro. In this study, we sought to assess the relative importance of epidermal growth factor (EGF) stimulation and c Src overexpression in conferring an invasive phenotype. METHODS: Breast carcinoma cells and colon epithelial cells which naturally express low levels of c-Src were used for these studies. The cells were transfected so that they overexpressed c Src; the mock-transfected parent lines were used as controls. Transfectants were tested for changes in invasion patterns after Src inhibition and EGF stimulation. RESULTS: Invasion assays in both cell systems confirmed the importance of c-Src in determining invasive potential. A significant correlation was shown between c Src kinase protein and cell invasion. Furthermore, Src inhibition significantly inhibited invasion in a dose-dependent manner. To clarify the relative contribution of EGF and c-Src to cell invasion, the ability of cells to invade through growth-factor-reduced matrigel, with or without EGF added, was compared to invasion through intact matrigel. The breast and colon cell lines behave quite differently in this regard. In the colon model, overexpressed c-Src is critical for cell invasion and stimulation with EGF is synergistic with c-Src overexpression. Conversely, the breast carcinoma cells transfected with c-Src were unable to invade without EGF stimulation and did not demonstrate the same synergistic relationship between c-Src and EGF. Instead, our results indicate that in BT474 breast carcinoma cells, EGF can substitute for c-Src in promoting breast cancer cell invasion. CONCLUSIONS: Because most breast carcinomas overexpress c-Src, it behooves one to question the extent to which reducing the amount of EGF and consequent EGFR activity will decrease invasion. In this study, the effects of EGF on cell invasion were determined in light of a single alteration in c-Src expression. Our results show that EGF enhances the impact of c-Src overexpression on invasion. In breast cancer cells, EGF is capable of inducing invasion to the same extent as c-Src overexpression. This suggests that anti-EGFR therapies will be efficacious in retarding breast carcinoma invasion and metastasis. PMID- 12643854 TI - Prognostic models of abdominal wound dehiscence after laparotomy. AB - BACKGROUND: Portions of the prospective, multi-institutional National Veterans Affairs Surgical Quality Improvement Program were used to develop and validate a perioperative risk index to predict abdominal wound dehiscence after laparotomy. METHODS: Perioperative data from 17,044 laparotomies resulting in 587 (3.4%) wound dehiscences performed at 132 Veterans Affairs Medical Centers between October 1, 1996, and September 30, 1998, were used to develop the model. Data from 17,763 laparotomies performed between October 1, 1998, and September 30, 2000, resulting in 562 (3.2%) dehiscences were used to validate the model. Models were developed using multivariable stepwise logistic regression with preoperative, intraoperative, and postoperative variables entered sequentially as independent predictors of wound dehiscence. The model was used to create a scoring system, designated the abdominal wound dehiscence risk index. RESULTS: Factors contributing significantly to the model and their point values (in parentheses) for the risk index include CVA with no residual deficit (4), history of COPD (4), current pneumonia (4), emergency procedure (6), operative time greater than 2.5 h (2), PGY 4 level resident as surgeon (3), clean wound classification (-3), superficial (5), or deep (17) wound infection, failure to wean from the ventilator (6), one or more complications other than dehiscence (7), and return to OR during admission (-11). Scores of 11-14 are predictive of 5% risk of dehiscence while scores of >14 predict 10% risk. CONCLUSIONS: This abdominal wound dehiscence risk index identifies patients at risk for dehiscence and may be useful in guiding perioperative management. PMID- 12643855 TI - Characteristics of bariatric surgery in an integrated VA Health Care System: follow-up and outcomes. AB - BACKGROUND: Since the 1991 NIH consensus conference, obesity surgery has been increasingly accepted as a form of therapy for morbid obesity. Approximately 40% of Veteran patients are obese and would potentially benefit from the operations. METHODS: Records were reviewed for all obesity operations performed at the Veterans Administration Greater Los Angeles Health Care System between January 1997 and April 2002. Morbidity, mortality, weight loss, and extent of follow-up were the outcomes measures assessed. RESULTS: Forty-six [11 Female (24%), 35 Male (76%)] patients underwent Roux-Y gastric bypass during the 63-month period we reviewed. There was one death from pulmonary hypertension unexpectedly encountered in the operating room. There was a single major complication: an anastomotic leak successfully treated with intravenous antibiotics. The Computerized Patient Record System contained extensive weight loss information, with dozens of weight measurements for these patients before and after surgery. Weight loss was sustained in all but 2 patients during the follow-up period. The only patients lost to follow-up were those referred from medical centers outside the boundaries of our integrated health care system. CONCLUSION: (1) The VA population has the opposite male/female ratio of populations reported in most weight loss studies. Because the health risks attributable to obesity are greater in males, the VA represents an important population to study that may benefit significantly from weight loss surgery. (2) Extensive clinical information available in the computerized medical record combined with frequent accession of health care resources by our patients resulted in a database rich in follow-up data for a population where long-term outcomes are traditionally difficult to obtain. (3) There was very low surgical morbidity and mortality in a high-risk population. This contrasts with the results of most volume-outcome studies and occurred in a low-volume hospital by a high-volume surgical and medical team. (4) Distinct patterns of weight loss were observed. PMID- 12643856 TI - Ascorbic acid dynamics in the seriously ill and injured. AB - BACKGROUND: In addition to the known beneficial effects of ascorbic acid on wound healing and the immune response, it is also a potent extracellular antioxidant. Recent work in septic rats suggests that high-dose ascorbic acid total parenteral nutrition (TPN) supplementation may protect cells from free radical injury and improve survival. In this study, we determined ascorbic acid levels in the immediate post-injury/illness period and evaluated the ability of early short term high levels of ascorbic acid in TPN to normalize plasma levels. MATERIALS AND METHODS: Ascorbic acid levels were determined in 12 critically injured patients and 2 patients with severe surgical infections. Each patient received TPN supplemented with increasing doses of ascorbic acid over a 6-day period. Therapeutic responses were determined by plasma and urine measurements using high pressure liquid chromatography. RESULTS: The initial mean +/- SEM baseline plasma ascorbic acid concentration was depressed (0.11 +/- 0.03 mg/dl) and unresponsive following 2 days on 300 mg/day supplementation (0.14 +/- 0.03; P = 1.0) and only approached low normal plasma levels following 2 days on 1000 mg/day (0.32 +/- 0.08; P = 0.36). A significant increase was noted following 2 days on 3000 mg/day (1.2 +/- 0.03; P = 0.005). CONCLUSION: We confirmed extremely low plasma levels of ascorbic acid following trauma and infection. Maximal early repletion of this vitamin requires rapid pool filling early in the post-injury period using supraphysiologic doses for 3 or more days. PMID- 12643857 TI - Effects of oral arginine and glutamine on radiation-induced injury in the rat. AB - BACKGROUND: Exposure of the abdominal region to ionizing radiation is associated with serious untoward symptoms of intestinal dysfunction and some reports indicate that nutrient supplements may reduce these adverse effects. This study was designed to investigate the possible beneficial effects of oral arginine or glutamine supplementation on the radiation-induced tissue injury. MATERIALS AND METHODS: Rats were given one of three feeding regimens: standard diet and water (control group), diet and water containing 2% arginine (arginine group), diet and water containing 2% glutamine (glutamine group) for 3 days prior to radiation. All rats were then subjected to a single does of 1100 cGy to the abdomen. Several serum biochemical parameters and the histologic alterations in different segments of gastrointestinal tract and liver were measured 4 days after irradiation. RESULTS: All the arginine-fed rats developed diarrhea on Day 4 postirradiation, compared to 71% incidence in control rats and 86% in glutamine-fed rats. Serum levels of aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) in the arginine group were markedly higher than those in other groups. On histological examination, radiation caused more serious damage to various segments of intestine in the arginine-fed rats compared to rats on other feeding regimens. CONCLUSION: These observations seriously question the beneficial effects of arginine and glutamine supplementations on radiation-induced tissue injury. PMID- 12643858 TI - Reduction of hepatic ischemia/reperfusion-induced injury by a specific ROCK/Rho kinase inhibitor Y-27632. AB - BACKGROUND: The low-molecular-weight GTPase Rho is known to act as a molecular switch by activating several downstream effectors, one of which is Rho-associated coiled-coil forming protein kinase (ROCK). ROCK/Rho kinase mediates cytoskeleton dependent cell functions, such as actomyosin-based smooth muscle contraction and integrin-mediated cell adhesion. A specific inhibitor of ROCK/Rho kinase, Y 27632, was recently developed. The present study examined whether Y-27632 could provide a beneficial effect on hepatic ischemia/reperfusion (I/R)-induced injury through the attenuation of microcirculatory disturbance. MATERIALS AND METHODS: In male Sprague-Dawley rats, normothermic partial ischemia was induced by clamping the hepatic pedicle to the left and median lobes for 90 min, followed by 2 h of reperfusion. In the treatment group, Y-27632 was intravenously administered prior to ischemic insult. Hepatic microcirculation was investigated by using intravital fluorescence microscopy. Liver enzyme release and histological changes of the liver tissue were also evaluated. RESULTS: Y-27632 significantly improved sinusoidal perfusion and reduced the number of leukocytes sticking in hepatic sinusoids and adhering in postsinusoidal venules. The postischemic narrowing of both sinusoids and postsinusoidal venules was also markedly suppressed. Consequently, liver enzyme release was reduced and postischemic histological damage was suppressed. CONCLUSIONS: A specific ROCK/Rho kinase inhibitor, Y-27632, was useful to alleviate hepatic I/R-induced injury through ameliorating postischemic microcirculation. The administration of Y-27632 may be a novel strategy for conquering hepatic I/R-induced injury. PMID- 12643859 TI - Differential salutary effects of nonselective and selective COX-2 inhibitors in postoperative ileus in rats. AB - BACKGROUND: Postoperative ileus (PI) is a common surgical complication, the treatment of which consists of supportive measures. AIM: The effects of several cyclooxygenase (COX) inhibitors and their interaction with L-arginine/nitric oxide synthase (NOS) pathway were tested in a rat PI model. METHODS: Intestinal transit was measured as Evans blue migration after skin incision, laparotomy, or laparotomy followed by evisceration and gut handling. RESULTS: In contrast to a selective inducible NOS (iNOS) blocker, L-N(6)-(1-iminoethyl)lysine hydrochloride (L-NIL), N(omega)-nitro-L-arginine methyl ester (L-NAME) reversed the additional inhibitory effects of gut manipulation after laparotomy on the gastrointestinal transit (GI) in a dose-dependent, L-arginine-sensitive manner. Laparotomy and manipulations of small intestine increased blood plasma nitrites and nitrates level (NOx), an effect preventable by L-NAME. Indomethacin, resveratrol (selective COX-1 blocker), and COX-2 antagonists, nimesulide, NS-398, DuP-697, and L-752860, attenuated the additional inhibitory effects of gut manipulation following laparotomy in a dose-dependent manner. In contrast, only nimesulide, NS 398, DuP-697, and L-752860 partly, but significantly, reversed the effects of laparotomy on the intestinal transit. Administration of L-NAME subsequent to COX inhibitors abolished the salutary effects of the latter, implying that at least the synthesis of either NO or prostanoids must remain unaffected to enable a return of GI transit during the postoperative period. CONCLUSION: In addition to NO synthesized by constitutive NOS (cNOS), prostaglandins produced by both COX-1 and COX-2 participate in the pathogenesis of PI, albeit in different pathological mechanisms. Thus laparotomy stimulated COX-2 activity, whereas gut manipulation led to an excessive cNOS activity and prostaglandin synthesis by COX-1. PMID- 12643860 TI - Popliteal vascular injury during total knee arthroplasty. AB - BACKGROUND: Popliteal vascular injury during total knee arthroplasty (TKA) is rare. Although the etiology, management, and outcome of this injury have not been systematically studied, several anecdotal reviews and small case series have suggested a high morbidity, with tourniquet injury as a predominant etiology. METHODS: We surveyed 190 members of a major regional vascular society concerning their experiences with popliteal vascular injuries sustained during TKA, specifically regarding the presentation, mechanisms of injury, diagnostic studies, surgical exposure and methods of repair, and outcomes. RESULTS: Thirteen surgeons provided data (6.8% response rate) on 19 patients with TKA-related popliteal artery injuries. In contrast to the historical literature, sharp, direct trauma was the predominant etiology of vascular injury. Vascular reconstructive techniques varied, with arterial bypass being the most common. There was no mortality in this series. Eighty-four percent (16/19) of the patients had full recoveries. Limb loss occurred in 2 of the 19 patients (10.5%). CONCLUSION: Popliteal vascular injuries during TKA are primarily the result of direct trauma to the vessel. Prompt diagnosis, extensile vascular exposure through a medial knee surgical incision, and standard reconstructive techniques offer a good prognosis for this limb-threatening complication. PMID- 12643861 TI - Adrenomedullin and adrenomedullin binding protein-1: their role in the septic response. AB - Adrenomedullin (AM) is a recently discovered, potent vasodilatory peptide with activities including maintenance of cardiovascular and renal homeostasis. Studies have indicated that AM is important in initiating the hyperdynamic response during the early stage of sepsis, and reduction of the vascular effects of AM marks the transition from the initial hyperdynamic phase to the late hypodynamic phase in experimental sepsis. The decreased AM responsiveness in late sepsis may be related to alterations in the AM receptor binding characteristics and/or signaling pathways. Genetic experiments have provided useful information by enhancing AM gene expression. Moreover, a plasma protein which binds AM, adrenomedullin binding protein-1 (AMBP-1), was reported very recently and is just beginning to be investigated as an important modulator in the biphasic septic response. In this regard, our recent results have demonstrated that AMBP-1 synergistically enhanced AM-induced vascular relaxation in both sham and septic animals. It appears that decreased levels of AMBP-1 play a critical role in producing vascular AM hyporesponsiveness during the late stage of sepsis. Furthermore, administration of AM and AMBP-1 in combination prevented the transition from the hyperdynamic to hypodynamic response during the progression of polymicrobial sepsis. Thus, modulation of vascular responsiveness to AM by AMBP-1 may provide a novel approach for the management of sepsis. PMID- 12643862 TI - HIV-associated lipodystrophy syndrome: description and pathogenesis. PMID- 12643864 TI - Assessment of antibiotic prescription in hospitalised patients at a Chinese university hospital. AB - OBJECTIVE: To describe and understand antibiotic prescribing in hospitalised patients in a university hospital. METHODS: Retrospective study using clinical data on antibiotic prescriptions in 1025 inpatient cases chosen from 21,000 inpatients in our hospital in 1997 by systematic sampling. RESULTS: Antibiotics were prescribed to 77.8% of inpatients, among which 55.2% were prescribed two or more kinds of antibiotics. In 58.5% of cases, antibiotic prescriptions were given therapeutically. Only 39 specimens in 1025 cases were taken for microbiological investigation. CONCLUSIONS: Few microbiological investigations were undertaken prior to antibiotic prescribing. The main challenges in the antibiotic prescribing are to make rational choices for antibiotic use and recognise the potential problems of abuse of antibiotics. PMID- 12643863 TI - Candida tropicalis fungemia in a tertiary care hospital. AB - Candida tropicalis is a frequent cause of fungemia in hospitals in Latin America. Candida albicans (33%) was the most frequently isolated species, followed by Candida parapsilosis (27%), and Candida tropicalis (24%) in tertiary care hospital in Brazil. We identified and retrospectively reviewed 27 cases of C. tropicalis fungemia that occurred at Hospital de Clinicas de Porto Alegre from 1996 to 1999. The mean age of the patients was 32 years (range 6 months to 88 years). Eight patients (29.6%) had hematological malignancy, and four (14.8%) had solid tumors. All the patients were taking broad-spectrum antibiotics, including vancomycin for at least 7 days. Antibiotics were given through a central venous catheter for the majority of the patients (77.7%). Relevant risk factors for candidemia in our patients included neutropenia (59.2%), and use of corticosteroids (37.0%) or cytotoxic drugs (40.7%). The onset of fever was the most frequent clinical manifestation (92.5%) of fungemia. Most of the patients (81.4%) were treated with amphotericin B or fluconazole. Overall mortality was 48.1%, and 7 (53.4%) of 13 deaths occurred within 10 days of the detection of candidemia. Results of the in vitro susceptibility testing of nine isolates of C. tropicalis from seven patients did not show resistance to fluconazole and amphotericin B.C. tropicalis presents as an important cause of fungemia in oncological and nononcological patients with central venous catheters taking broad-spectrum antibiotics. Although there was no evidence of resistance of C. tropicalis to amphotericin B and fluconazole, patients treated with antifungal agents presented with a high mortality rate in the hospital setting. PMID- 12643866 TI - Relevance and complications of intravenous infusion at the emergency unit at Nice university hospital. AB - OBJECTIVE: Insertion of peripheral venous catheters (PVCs) is current practice within the hospital environment and particularly in the emergency department (ED). This simple yet invasive technique may result in venous thrombosis, infection or mechanical complications. We conducted a prospective study at the Nice University Hospital ED in order to determine the frequency, relevance and complications of peripheral venous cannulation. PATIENTS AND METHODS: Fourteen 24 h periods were surveyed over the months of May and June 2000, during which 2515 patients over 16 years of age attended the unit. Demographic and medical data were recorded for every patient who received a PVC. These patients were followed at 12 h intervals until the catheter was removed. Reasons for PVC, time left in place, and eventual complications were recorded. Justification for PVC was evaluated upon arrival at the ED, upon exiting the ED and in some cases within the hospital ward. RESULTS: Six hundred and thirty of 2515 patients (25%) received a PVC (290 women (46%) and 340 men (54%); meanage 58 years). Indication for the PVC was considered unjustified in 24.8% of cases upon arrival at the ED, and 33.8% upon leaving the ED. Upon admission in a hospital ward after passing through the ED, out of 318 patients, the PVC was left in place for no reason in 63 (20%). Overall, 390 PVCs were followed until the time of their removal. Mean duration of IV infusion was 28 h. Among these 390 patients, 62 (15.9%) developed complications, of which 54 (13.6%) had thrombophlebitis and 9 (2.3%) developed local infection. Mean duration of PVC left in place for patients with complications was 50 h vs 25 h for patients with no complications (P<0.001). CONCLUSION: Insertion of a PVC is common practice especially in EDs. The excessive use of this procedure leads to extra cost and iatrogenic complications. A renewed definition of its indication and raised awareness among hospital staff concerning the risks involved with this standard procedure should result in less use of PVC and fewer complications. PMID- 12643865 TI - Does infection with Human Immunodeficiency Virus affect the antibody responses to Plasmodium falciparum antigenic determinants in asymptomatic pregnant women? AB - OBJECTIVES: HIV-seropositive pregnant women are more susceptible to malaria than HIV-seronegative women. We assessed whether HIV infection alters maternal and cord plasma malarial antibody responses and the mother-to-infant transfer of malaria antibodies. METHODS: We determined plasma levels of maternal and cord antibodies [Immunoglobulin (IgG)] to recombinant malarial proteins [merozoite surface protein 1 (MSP-1(19kD)), the erythrocyte binding antigen (EBA-175)], the synthetic peptides [MSP-2, MSP-3, rhoptry associated protein 1 (RAP-1), and the pre-erythrocytic stage, circumsporozoite protein (NANP)(5)] antigenic determinants of Plasmodium falciparum; and tetanus toxoid (TT) by ELISA among samples of 99 HIV-seropositive mothers, 69 of their infants, 102 HIV-seronegative mothers and 62 of their infants. RESULTS: The prevalence of maternal antibodies to the malarial antigenic determinants ranged from 18% on MSP3 to 91% on EBA-175; in cord plasma it ranged from 13% to 91%, respectively. More than 97% of maternal and cord samples had antibodies to TT. In multivariate analysis, HIV infection was only associated with reduced antibodies to (NANP)(5) in maternal (P=0.001) and cord plasma (P=0.001); and reduced mother-to-infant antibody transfer to (NANP)(5) (P=0.012). This effect of HIV was independent of maternal age, gravidity and placental malaria. No consistent HIV-associated differences were observed for other antigenic determinants. CONCLUSION: An effect of HIV infection was only observed on one malarial antigenic determinant, suggesting that the increased susceptibility to malaria among HIV-infected pregnant women may not be explained on the basis of their reduced antibody response to malaria antigens. PMID- 12643867 TI - Long-term follow up study of vertical HTLV-I infection in children breast-fed by seropositive mothers. AB - Infection of Human T-lymphotropic virus type-I (HTLV-I) was investigated by long term follow up surveys of mother's milk-fed infants. HTLV-I infections of infants via seropositive mother's milk, that is, anti-HTLV-I antibody-positive infants, increased in number up to the age 2, but no infants became antibody-positive thereafter. Infants who had became antibody positive by age 2 remained so at age 11-12. HTLV-I infection via feeding with mother's milk was established by the age 2. While in epidemiologic surveys an increase of the anti-HTLV-I antibody positive rate has been reported, this survey revealed that after acquisition of HTLV-I from breast feeding, there was no further horizontal transmission prior to puberty. PMID- 12643868 TI - Investigation of tropical eosinophilia; assessing a strategy based on geographical area. AB - OBJECTIVES: Patients with eosinophilia are an important clinical problem. This study aimed to assess the most efficient manner of investigating patients with peripheral eosinophilia (eosinophil count >0.5x10(9)ml(-1)) presenting from the tropics. METHODS: Patients attending the Hospital for Tropical Diseases, London, from October 1997 to March 2002 for investigation of eosinophilia were identified prospectively. Laboratory, clinical and demographic data were recorded from laboratory and clinical records. An investigation set was proposed prospectively and assessed for all geographical areas (stool microscopy, strongyloides culture and serology), all of Africa (additional schistosomal serology, terminal urine microscopy and filarial serology) and West Africa (additional day-bloods for microfilaria). RESULTS: Data was analysed for 261 patients. At least one helminthic cause for eosinophilia was found in 64% of patients (median eosinophilia 1.2x10(9)ml(-1)). Seventeen per cent of patients had more than one helminth species found. Median eosinophilia increased with number of diagnoses per patient. The proposed investigation sets were validated, with high yield for all proposed tests apart from filarial serology outside West Africa, and good sensitivity. CONCLUSIONS: Initial investigation of eosinophilia in patients presenting from the tropics may be guided by a simple investigation set depending on broad area of travel which has high sensitivity and yield. Patients frequently have more than one helminthic cause of eosinophilia. PMID- 12643869 TI - Unusual sequelae of meningococcal infection: a case report of hepatic dysfunction, protracted neurological complications and late cutaneous manifestations associated with serogroup y. PMID- 12643870 TI - Infective tricuspid valve endocarditis with septic pulmonary emboli due to puncture of an endogenous arteriovenous fistula in a chronic hemodialysis patient. AB - BACKGROUND: Patients on chronic hemodialysis are at high risk for infectious endocarditis due to prosthetic access devices such as cuffed venous catheters and polytetrafluoroethylene grafts. Right-sided endocarditis without any predisposing factors is rare in dialysed patients. CASE REPORT: A 42-year-old man with chronic renal failure was referred to hospital due to febrile temperatures. Hemodialysis was performed via an autogenous arteriovenous fistula. He had neither any recognized underlying valve disease nor any percutaneous interventions previously. Chest radiography and computed-tomography revealed numerous lung abscesses caused by tricuspid endocarditis detected by transesophageal echocardiography. Antibiotics were given for 5 weeks curing the infection without necessity of surgical treatment. CONCLUSIONS: Primary arteriovenous fistulas have the lowest rates of infections and are the access of choice for chronic hemodialysis patients. However, repeated vascular punctures even of autogenous grafts expose dialysis patients to bacteremia and imply a higher risk of infectious endocarditis. PMID- 12643871 TI - Haemophilus paraphrophilus prosthetic valve endocarditis. AB - We report a case of prosthetic, aortic valve, infective endocarditis caused by Haemophilus paraphrophilus. There are no other cases described in the available literature where this microorganism has caused prosthetic valve endocarditis and no other case reported involving only the aortic valve. PMID- 12643872 TI - Rapidly fatal Haemophilus influenzae serotype f sepsis in a healthy child. AB - A previously healthy 4-year-old child became acutely ill with vomiting and low grade fever. The following day she suddenly became limp and unresponsive. She experienced acute septic shock and despite aggressive treatment died. Blood cultures grew ampicillin-resistant Haemophilus influenzae type f. There was no evidence of bacterial pneumonia or meningitis. To our knowledge, this represents the first case of fatal H. influenzae type f sepsis in a child without an identifiable focus or underlying predisposing condition. Despite the overwhelming success of the H. influenzae type b vaccine, physicians need to be aware of the potential for severe and fatal H. influenzae infections other than type b. PMID- 12643873 TI - Late onset Ommaya reservoir infection due to Staphylococcus aureus: case report and review of Ommaya Infections. AB - The Ommaya reservoir system has been used for the treatment of chronic central nervous system infections and intracranial tumors for more than three decades. The majority of reported Ommaya reservoir infections occur proximate to the time the device is accessed. A review of the literature reveals that late onset of reservoir infection is quite rare. We report a case of Ommaya reservoir infection due to Staphylococcus aureus that was diagnosed seven years after its insertion and usage for intracerebral non-Hodgkin's lymphoma and review the literature on the microbiology and management of Ommaya reservoir infections. PMID- 12643874 TI - Multi-system Infection with Nocardia farcinica-therapy with linezolid and minocycline. AB - We describe a case of disseminated infection with a multiple-resistant strain of Nocardia farcinica, probably resulting from direct inoculation during a road traffic accident. Initial presentation was with pulmonary symptoms, with subsequent development of cutaneous, renal, soft tissue and cerebral involvement. Precise microbiological diagnosis was delayed. Once sensitivity test results were available, the patient was stabilised on linezolid and minocycline. Premature withdrawal of therapy at one month resulted in recrudescence of infection, requiring re-institution of treatment. Linezolid was discontinued after a total of 4 months, because of evidence of myelosuppression and visual impairment, which subsequently improved. Monotherapy with minocycline was continued for a total of 12 months. The patient now remains well. PMID- 12643875 TI - Isolation and molecular confirmation of a multiresistant catalase-negative Staphylococcus aureus in Nigeria. PMID- 12643876 TI - The safety and tolerability of switching from a non-failing antiretroviral regimen to lopinavir. PMID- 12643877 TI - The LIFE study: the straw that should break the camel's back. PMID- 12643878 TI - Management of non ST-segment elevation acute coronary syndromes-continuing the search for the bad guys. PMID- 12643879 TI - Who wants his plaque sealed? PMID- 12643880 TI - Hypertrophic cardiomyopathy-the clinical challenge of managing a hereditary heart condition. PMID- 12643881 TI - Fibrinogen: a predictor of stroke and marker of atherosclerosis. PMID- 12643882 TI - Well kept secrets of the genome. PMID- 12643883 TI - Outcome beyond blood pressure control? PMID- 12643885 TI - Does the discharge ECG provide additional prognostic insight(s) in non-ST elevation ACS patients from that acquired on admission? AB - BACKGROUND: Although the prognostic value of admission ST changes in patients with non-ST elevation acute coronary syndrome (ACS) is established, the utility of the discharge ECG is unknown. Accordingly, using the PARAGON-B Troponin substudy, we assessed the prevalence of ST depression on both admission and discharge ECG, the likelihood of developing new Q-waves at discharge and the additional prognostic value of these changes. METHODS AND RESULTS: Nine hundred and eighteen patients were studied; 542 patients (59%) had admission ST downward arrow > or =1mm and 376 patients (41%) did not and their 6-month mortality was 4.4 vs 0.8%, P=0.002, respectively. Of patients with ST downward arrow on admission, 320 (59%) normalized their ST segment at discharge. Of patients without ST downward arrow on admission, 35 (9.3%) developed new ST downward arrow at discharge. Patients with persistent ST downward arrow on discharge had a higher 6-month mortality (6.0 vs 0.9%), (re)MI (16.3 vs 7.4%), and death/(re)MI (20.0 vs 8.3%) than those who never had ST downward arrow (all P< or =0.002). Two hundred and fifty-six patients had Q-waves on admission whereas by discharge 320 had Q-waves. Patients with Q-waves on discharge vs those without had a higher mortality (4.8 vs 1.9%), (re)MI (13.8 vs 8.3%), and death/(re)MI (16.4 vs 9.6%) at 6 months (all P< or =0.021). CONCLUSIONS: This study highlights that the dynamic ECG changes which occur between admission and discharge in non-ST elevation ACS patients allows further risk stratification in determining the likelihood of 6-month death and/or re(MI). PMID- 12643884 TI - Acute infections in children are accompanied by oxidative modification of LDL and decrease of HDL cholesterol, and are followed by thickening of carotid intima media. AB - BACKGROUND: Atherosclerosis begins early in life. Infections might contribute to the pathogenesis of atherosclerosis. In this study, we investigated whether acute infections in children could alter the carotid wall morphology and the lipid profile. METHODS: Mean carotid intima-media thickness (IMT) was measured by high resolution ultrasound in 28 hospitalised children (mean age: 5+/-2 years), who fulfilled the diagnostic criteria of acute infections (body temperature, >38 degrees C; C-reactive protein, >15mg/ml, and clinical), and in 20 age- and gender matched controls. Antibodies against oxidised low-density lipoprotein (anti-oxLDL antibodies), as well as total and high-density lipoprotein cholesterol (HDL-C) were analysed in all children. The infection group was investigated both during the acute illness and 3 months after clinical recovery (post-infection). RESULTS: During the acute illness, the infection group had elevated anti-oxLDL antibodies and decreased HDL-C, as compared to those obtained at 3 months and in controls (p<0.05). These changes in the infection group were followed, at 3 months, by thickening of carotid intima-media. Those who received antibiotics during their acute illness had less carotid thickening than those who were not treated with antibiotics (p<0.05). CONCLUSION: Acute infections in children seem to be accompanied by enhanced oxidative modification of LDL and by decrease in HDL-C. These lipid changes may be followed by thickening of carotid artery intima-media. These findings suggest that, in childhood, acute infections could be associated with increased risk of atherosclerosis, and warrant further studies on this topic. PMID- 12643886 TI - The value of routine non-invasive tests to predict clinical outcome in stable angina. AB - BACKGROUND: Chronic stable angina is a common condition, but considerable differences exist in the likelihood of acute coronary events such as CHD death, non-fatal myocardial infarction (MI) and unstable angina between individual patients. Effective risk prediction is necessary for optimum management. The aim of this study was to identify clinical features and non-invasive test parameters associated with high risk of these coronary events in stable angina and compose a clinically useful model to predict adverse outcomes in this population. METHODS: Six hundred and eighty-two patients with stable angina and a positive exercise test (1mm ST depression) from the Total Ischaemic Burden European Trial (TIBET) study, were studied. Resting ECG, exercise tolerance testing and echocardiography were performed at baseline, off anti-anginal therapy. The patients were then randomised to treatment with atenolol, nifedipine or a combination of both. Clinical follow up continued for an average of 2 years (range 1-3 years). RESULTS AND CONCLUSIONS: Prior MI or prior CABG were the clinical parameters associated with adverse outcome in patients with stable angina and a positive exercise test. On the ECG, left ventricular hypertrophy was predictive, and on echocardiogram, increased left ventricular dimensions were predictive of adverse events. When combined with time to ischaemia on exercise testing in a simple clinically applicable table these factors could be used to predict of 2 year probability of events for an individual patient. PMID- 12643887 TI - Clinical and angiographic outcome of patients with mild coronary lesions treated with balloon angioplasty or coronary stenting. Implications for mechanical plaque sealing. AB - AIMS: To investigate the clinical and angiographic outcome of patients with mild coronary lesions treated with balloon angioplasty or coronary stenting (coronary plaque sealing, i.e. dilatation of angiographically non-significant lesions) compared to moderate and severe stenoses. METHODS AND RESULTS: Patients with chronic stable angina and a single de novo lesion in a native coronary vessel scheduled to undergo percutaneous coronary intervention (PCI) were selected from 14 different studies. Off-line analysis of angiographic outcomes was assessed in all patients using identical and standardised methods of data acquisition, analysis and definitions. Clinical endpoints were adjudicated by independent clinical events committees. All quantitative coronary angiographic (QCA) analyses were performed in the same core laboratory. Stenosis severity prior to PCI was categorised into three groups: <50% diameter stenosis (DS), 50-99%DS and >99%DS pre. A total of 3812 patients were included in this study; 1484 patients (39%) were successfully treated with balloon angioplasty (BA) only and stented angioplasty was performed in 2328 patients (61%).One-year mortality and rate of non-fatal myocardial infarction (MI) (Kaplan-Meier) did not differ between BA and stented angioplasty for any of the stenosis severity categories. Following BA, the combined event rate (death and non-fatal MI) was 4.8, 4.6 and 0% in the <50, 50-99 and >99%DS categories, respectively. Following stented angioplasty, the combined event rate was 3.1, 4.4 and 4.8% in the same categories. The need for repeat revascularisation corrected for stenosis severity in the Cox proportional hazards regression model was reduced by 20% after stented angioplasty (hazard ratio (HR) 0.80, 95%CI 0.69-0.93). CONCLUSION: The concept of plaque sealing is appealing from the theoretical point of view. However, with current technology, plaque sealing cannot prevent death and future non-fatal MIs in the long-term because 1-year event rates after PCI of non-significant stenoses remain unacceptably elevated when compared with the estimated 1-year probability of a non-fatal MI in lesions with a <50%DS. Moreover, major adverse cardiac events at 1-year after PCI are not directly related to the degree of pre-procedural stenosis severity. PMID- 12643888 TI - Bisoprolol dose-response relationship in patients with congestive heart failure: a subgroup analysis in the cardiac insufficiency bisoprolol study(CIBIS II). AB - AIMS: Whether all patients with congestive heart failure (CHF) need to reach the target dose of beta-blocker to obtain a benefit in terms of survival remains uncertain. METHODS AND RESULTS: We classified by tertile the 2647 patients enrolled in CIBIS II according to the last tolerated dose: low dose (LD: 1.25, 2.5 or 3.75mg/day, n=434), moderate dose (MD: 5 or 7.5mg/day, n=328) and high dose (HD: 10mg/day, n=565) of bisoprolol or placebo (LD=234, MD=278 and HD=808). In both groups, patients tolerating only low doses were significantly older with more severe New York Heart Association (NYHA) functional class and higher frequency of co-morbidities. Treatment withdrawal was associated with a significant increase of mortality in the bisoprolol group (relative hazard (RH)=2.13, 95% confidence interval (CI)=1.43-3.17, p=0.0002). After adjustment, all-cause mortality was significantly reduced in the bisoprolol group compared to placebo regardless of the dose level considered: LD (RH=0.66, 95% CI=0.48-0.92), MD (RH=0.33, 95% CI=0.21-0.51) or HD (RH=0.59, 95% CI=0.40-0.89). CONCLUSIONS: Bisoprolol reduces mortality in CHF patients at all tolerated dose levels and its withdrawal increases the risk of mortality. Efforts should be made to maintain bisoprolol therapy based on the individual patient's tolerability. PMID- 12643889 TI - Genetic knowledge and counselling skills of Dutch cardiologists: sufficient for the genomics era? AB - AIMS: Genetic scientific knowledge is growing rapidly but how this affects clinical practice is unclear. We investigated the levels of knowledge, practical skills and clinical genetic practices of Dutch cardiologists. METHODS AND RESULTS: A survey was designed to assess cardiologists' experience with genetic aspects of hypertrophic cardiomyopathy, self-reported genetic knowledge, and genetic skills in general and aimed at this disease. In addition, cardiologists' opinions on five possible measures for improvement were obtained. STUDY POPULATION: all Dutch cardiologists (n=643). Median number of patients suffering from hypertrophic cardiomyopathy per cardiologist is five. Forty-one percent of respondents do not give information about genetics to all their patients. Cardiologists rarely initiate DNA tests for hypertrophic cardiomyopathy. Only 38% refer patients to clinical geneticists. Self-reported knowledge levels are low (average score 3.3-5.1, 0-10 scale). Cardiologists with an established working relationship with a clinical geneticist report significantly higher levels of knowledge. Clinical guidelines, education and improved collaboration with clinical geneticists are preferred. CONCLUSION: Dutch cardiologists' genetic knowledge and clinical genetic practice levels are insufficient. As a result, clinical genetic care for patients with hypertrophic cardiomyopathy is poor. Improvements proposed include advancement of knowledge (education, professional guidelines) and structural measures (working relationships, multidisciplinary outpatient clinics). Collaboration of cardiologists and clinical geneticists is urgently needed to optimise cardiogenetic patient care. PMID- 12643890 TI - Fibrinogen predicts ischaemic stroke and advanced atherosclerosis but not echolucent, rupture-prone carotid plaques: the Copenhagen City Heart Study. AB - AIMS: Whether the association between fibrinogen and cardiovascular events reflects an association with advanced atherosclerosis in general, or rupture prone plaques in particular, is unclear. We examined whether fibrinogen predicts incidence of ischaemic stroke, advanced atherosclerosis (measured as carotid artery stenosis) and/or echolucent, rupture-prone plaques. METHODS AND RESULTS: Study 1-8755 Copenhagen City Heart Study stroke-free participants; we observed 235 ischaemic strokes during 6 years of follow-up. Study 2-318 carotid stenosis patients and 1584 age- and gender-matched controls. Study 3-159 patients with echolucent vs 159 patients with echo-rich carotid artery plaques. Fibrinogen above vs below the median value of 3 g l(-1)predicted risk of ischaemic stroke (relative risk: 1.9; 95% CI: 1.4-2.5; 235 events). Significant risk was found in men (2.7; 1.7-4.2; 113 events) and with a similar trend in women (1.4; 0.9-2.0; 122 events), in young (5.2; 1.1-26; eight events) and middle aged (2.9; 1.6-5.4; 64 events) with a similar trend in the elderly (1.4; 1.0-2.0; 163 events). Fibrinogen levels in those with and without ischaemic stroke were 3.6 and 3.1 g l(-1)(ANCOVA: P<0.0001). Likewise, in those with and without carotid artery stenosis fibrinogen levels were 4.7 and 3.1 g l(-1)(P<0.0001); equivalent values for high-sensitive C-reactive protein were 3.6 and 1.4 mg l(-1)(P<0.0001). Finally, neither fibrinogen nor high-sensitive C-reactive protein levels differed between those with echolucent and echo-rich carotid artery plaques (P=0.61 and P=0.28); the power to exclude a 15% increase in fibrinogen or a 50% increase in high-sensitive C-reactive protein was 98 and 54%, respectively. CONCLUSIONS: Elevated fibrinogen predicts future ischaemic strokes, particularly in men and in the young and middle aged. This is most likely a reflection of advanced atherosclerosis, rather than an association with rupture-prone plaques. PMID- 12643891 TI - The genetic determination of left ventricular mass in healthy adults. AB - AIMS: The extent to which left ventricular (LV) mass, an independent cardiovascular risk factor, is determined by genetic factors is unclear. The aim of this study was to assess the heritability of LV mass and its association with three potential candidate genes. METHODS: A population-based adult twin study model was utilized. Echocardiographic assessment of LV mass was performed in 110 twin pairs (mean age 55.9+/-10.9 years). An estimate of genetic determination, heritability, was calculated for the main echocardiographic parameters. The cohort were genotyped for the G-protein beta-3, aldosterone synthase, and beta-1 adrenoceptor genes. RESULTS: The intra-class correlation coefficients for LV mass were 0.69 for monozygotic (r-MZ) twins and 0.32 for dizygotic (r-DZ) twins, P=0.008 (heritability estimate of 0.69). This pattern persisted following correction for known confounding factors. Within-pair differences in the monozygotic, discordant and concordant dizygotic twins showed no differences for the three genes with respect to left ventricular wall thickness or mass. There was a non-significant trend towards a relationship between LV mass and the beta-1 adrenoceptor genotype. CONCLUSION: Within a normal population left ventricular mass has a significant genetic determination. Further investigation of potential candidate genes is required. PMID- 12643892 TI - Simple reaction time and statistical facilitation: a parallel grains model. AB - A race-like model is developed to account for various phenomena arising in simple reaction time (RT) tasks. Within the model, each stimulus is represented by a number of grains of information or activation processed in parallel. The stimulus is detected when a criterion number of activated grains reaches a decision center. Using the concept of statistical facilitation, the model accounts for many classical effects on mean simple RT, including those of stimulus area, stimulus intensity, stimulus duration, criterion manipulations, redundant stimuli, and the dissociation between intensity effects on simple RTs and temporal order judgments. The model is also consistent with distributional properties of simple RTs. PMID- 12643893 TI - An evidential support accumulation model of subjective probability. AB - A model of cue-based probability judgment is developed within the framework of support theory. Cue diagnosticity is evaluated from experience as represented by error-free frequency counts. When presented with a pattern of cues, the diagnostic implications of each cue are assessed independently and then summed to arrive at an assessment of the support for a hypothesis, with greater weight placed on present than on absent cues. The model can also accommodate adjustment of support in light of the baserate or prior probability of a hypothesis. Support for alternatives packed together in a "residual" hypothesis is discounted; fewer cues are consulted in assessing support for alternatives as support for the focal hypothesis increases. Results of fitting this and several alternative models to data from four new multiple-cue probability learning experiments are reported. PMID- 12643894 TI - Developing a new quantitative account of backward masking. AB - A new general explanation for u-shaped backward masking is analyzed and found to predict shifts in the interstimulus interval (ISI) that produces strongest masking. This predicted shift is then compared to six sets of masking data. The resulting comparisons force the general explanation to make certain assumptions to account for the data. In this way, the experimental data promote the development of a new theory of backward masking. The new theory suggests interpretations of the data that are sometimes novel, often more precise, and sometimes contrary to interpretations that are prevalent in the literature. PMID- 12643896 TI - Inhibition of bacterial IF2 binding to fMet-tRNA((fMet)) by aminoglycosides. AB - Screening for inhibitors of bacterial protein synthesis Initiation Factor 2 (IF2) binding to N-formyl-Methionyl-transfer RNA (fMet-tRNA((fMet))) identified a series of aminoglycosides, that included amikacin and kanamycin A1, as inhibitors of this interaction. Subsequent testing revealed that aminoglycosides displayed a wide range of inhibitory activity. However, the failure of these compounds to completely inhibit binding of IF2 to fMet-tRNA((fMet)), the known ability of aminoglycosides to bind RNA, and the ability of the aminoglycosides to displace PicoGreen bound to fMet-tRNA((fMet)) suggest these compounds act by binding fMet tRNA((fMet)). This hypothesis is further supported by isothermal denaturation experiments that failed to show any interaction between the IF2 protein and the aminoglycosides. PMID- 12643897 TI - Synthesis of aza and oxaglutamyl-p-nitroanilide derivatives and their kinetic studies with gamma-glutamyltranspeptidase. AB - A new series of L-glutamic acid p-nitroanilide analogues has been synthesized and tested as substrates and inhibitors of rat kidney gamma-glutamyltranspeptidase (GGT). Kinetic parameters (K(m) and k(cat)) were determined for each analogue and provide insight into the scope and limits of GGT catalytic efficiency. PMID- 12643898 TI - Syntheses and biological properties of cysteine-reactive epibatidine derivatives. AB - The synthesis of epibatidine derivatives modified at the 2-position of the pyridine or pyrimidine rings by reactive functions are described for potential irreversible site-directed coupling reactions on cysteine mutants of neuronal nicotinic acetylcholine receptors. An improved synthesis of the 7 azabicyclo[2,2,1]hepta-2,5-diene key intermediate has been developed to allow reproducible syntheses of the epibatidine derivatives. Binding tests and electrophysiological experiments allowed to select the 2-substituted alpha chloroacetamido 13 and the chloropyrimidine derivative 11 as potential site directed probes for the epibatidine binding site. PMID- 12643899 TI - Carbonic anhydrase inhibitors: inhibition of the tumor-associated isozyme IX with aromatic and heterocyclic sulfonamides. AB - The inhibition of the tumor-associated transmembrane carbonic anhydrase IX (CA IX) isozyme has been investigated with a series of aromatic and heterocyclic sulfonamides, including the six clinically used derivatives acetazolamide, methazolamide, ethoxzolamide, dichlorophenamide, dorzolamide and brinzolamide. Inhibition data for the physiologically relevant isozymes I and II (cytosolic forms) and IV (membrane-bound) were also provided for comparison. A very interesting and unusual inhibition profile against CA IX with these sulfonamides has been observed. Several nanomolar (K(I)-s in the range of 14-50 nM) CA IX inhibitors have been detected, both among the aromatic (such as orthanilamide, homosulfonilamide, 4-carboxy-benzenesulfonamide, 1-naphthalenesulfonamide and 1,3 benzenedisulfonamide derivatives) as well as the heterocylic (such as 1,3,4 thiadizole-2-sulfonamide, etc.) sulfonamides examined. Because CA IX is a highly active isozyme predominantly expressed in tumor tissues with poor prognosis of disease progression, this finding is very promising for the potential design of CA IX-specific inhibitors with applications as anti-tumor agents. PMID- 12643900 TI - Prodrugs to enhance central nervous system effects of the TRH-like peptide pGlu Glu-Pro-NH2. AB - Potential prodrugs for the TRH-like tripeptide pGlu-Glu-Pro-NH(2) were synthesized either by esterifying the Glu side-chain of the parent peptide in solution with alcohols in the presence of resin-bound dicyclohexylcarbodiimide or by solid-phase peptide chemistry. Affinities of these ester prodrugs to lipid membranes as predictors of the transport across the blood-brain barrier were compared by immobilized artificial membrane chromatography, and prodrug activation was tested in the brain tissue of experimental animals. Esters of pGlu Glu-Pro-NH(2) with long-chain primary alcohols emerged as potentially useful prodrugs to improve the central nervous system activity of pGlu-Glu-Pro-NH(2) upon systemic administration, as revealed by the enhancement of analeptic activity in mice. PMID- 12643901 TI - N-Benzoyl amino acids as LFA-1/ICAM inhibitors 1: amino acid structure-activity relationship. AB - The association of ICAM-1 with LFA-1 plays a critical role in several autoimmune diseases. N-2-Bromobenzoyl L-tryptophan, compound 1, was identified as an inhibitor to the formation of the LFA-1/ICAM complex. The SAR of the amino acid indicates that the carboxylic acid is required for inhibition and that L histidine is the most favored amino acid. PMID- 12643902 TI - A novel type of fluorescent boronic acid that shows large fluorescence intensity changes upon binding with a carbohydrate in aqueous solution at physiological pH. AB - In this paper we report 8-quinolineboronic acid as a novel type of fluorescent probe for carbohydrates. This boronic acid responds to the binding of a carbohydrate with over 40-fold increases in fluorescence intensity and shows optimal fluorescence change at physiological pH in aqueous solution. PMID- 12643903 TI - Nonbenzamidine isoxazoline derivatives as factor Xa inhibitors. AB - Factor Xa (fXa) is an important serine protease in the blood coagulation cascade. Inhibition of fXa has emerged as an attractive target for potential therapeutic applications in the treatments of both arterial and venous thrombosis. Herein, we describe a series of non-benzamidine isoxazoline derivatives as fXa inhibitors. The chloroaniline group was found to be the most potent benzamidine mimic in this series. Chloroaniline 1 (ST368) has a K(i) value of 1.5 nM against fXa and is highly selective for fXa relative to thrombin and trypsin. PMID- 12643904 TI - Synthesis and binding affinity of neuropeptide Y at opiate receptors. AB - Neuropeptide Y and several metabolic fragments were synthesized and evaluated for binding affinity at non-selective opiate receptors. Neuropeptide Y and several C terminal fragments were shown to bind to non-selective opiate receptors with an affinity similar to that of Leu-enkephalin. PMID- 12643905 TI - Facile synthesis of 7-amino-4-carbamoylmethylcoumarin (ACC)-containing solid supports and their corresponding fluorogenic protease substrates. AB - The bifunctional fluorophore, 7-amino-4-carbamoylmethylcoumarin (ACC) without any protection groups, was regioselectively attached to different solid supports functionalized with a primary amino group. The resulting resins were used to synthesize fluorogenic protease substrates with high yield and purity. PMID- 12643907 TI - Synthesis and hybridization property of novel 2',5'-isoDNA mimic chiral peptide nucleic acids. AB - 2',5'-isoDNA mimic chiral peptide nucleic acid (isoPNA) monomers derived from D- and L-aspartic acids were synthesized. These novel monomers were incorporated in aminoethylglycine peptide nucleic acid (aegPNA) thymine dodecamers, and the hybridization properties to RNA and DNA were demonstrated by UV thermal denaturation. PMID- 12643906 TI - The newly synthesized linoleic acid derivative FR236924 induces a long-lasting facilitation of hippocampal neurotransmission by targeting nicotinic acetylcholine receptors. AB - The newly synthesized linoleic acid derivative, FR236924, induces a long-lasting facilitation of hippocampal neurotransmission based on a persistent enhancement in the activity of presynaptic nicotinic ACh receptors via a PKC pathway and the ensuing increase in glutamate release, not only in vitro but in vivo at a low dosage (2mg/kg, ip), which suggested the possibility of its use as a promising anti-dementia drug. PMID- 12643908 TI - Anastatins A and B, new skeletal flavonoids with hepatoprotective activities from the desert plant Anastatica hierochuntica. AB - New skeletal flavonoids, anastatins A and B, were isolated from the methanolic extract of an Egyptian medicinal herb, the whole plants of Anastatica hierochuntica. Their flavanone structures having a benzofuran moiety were determined on the basis of chemical and physicochemical evidence. Anastatins A and B were found to show hepatoprotective effects on D-galactosamine-induced cytotoxicity in primary cultured mouse hepatocytes and their activities were stronger than those of related flavonoids and commercial silybin. PMID- 12643909 TI - Substituted 4-methylquinolines as a new class of anti-tuberculosis agents. AB - We report synthesis and anti-tuberculosis activities of a series of novel ring substituted quinolines. The most effective compound of the series 3d (MIC=6.25 microg/mL, Mycobacterium tuberculosis H37Rv strain) was synthesized in one step; thus is an attractive lead molecule for anti-tuberculosis drug development. The results of this study represent the discovery of ring-substituted 4 methylquinolines as new class of potential anti-tuberculosis agents. PMID- 12643911 TI - Alkylation of manganese(II) tetraphenylporphyrin by antimalarial fluorinated artemisinin derivatives. AB - The alkylating properties of two artemisinin derivatives bearing a trifluoromethyl substituent at C10 were evaluated toward manganese(II) tetraphenylporphyrin, considered as a heme model. Chlorin-type covalent adducts were obtained by alkylation of the porphyrin ring by C-centered radicals derived from reductive activation of the peroxide function of the drugs. PMID- 12643910 TI - Identification of a novel series of selective 5-HT7 receptor antagonists. AB - Novel 5-HT(7) receptor antagonists containing the benzocycloheptanone core were identified from high throughput screening. Molecular modelling and SAR studies have converted these intractable hits into a more potent, selective and tractable series, exemplified by compound (25), SB-691673. PMID- 12643912 TI - alpha-Selective glycosylation with 5-thioglucopyranosyl donors; synthesis of an isomaltotetraoside mimic composed of 5-thioglucopyranose units. AB - A general method for alpha-selective glycosylation with 5-thioglucopyranosyl donors followed by efficient deprotection of the resulting products was developed. This methodology was utilized in the synthesis of an isomaltotetraoside analogue. PMID- 12643913 TI - The identification of clinical candidate SB-480848: a potent inhibitor of lipoprotein-associated phospholipase A2. AB - Modification of the pyrimidone 5-substituent in clinical candidate SB-435495 has given a series of inhibitors of recombinant lipoprotein-associated phospholipase A(2) with sub-nanomolar potency. Cyclopentyl fused derivative 21, SB-480848, showed an enhanced in vitro and in vivo profile versus SB-435495 and has been selected for progression to man. PMID- 12643914 TI - Biphenyls as potent vitronectin receptor antagonists. Part 2: biphenylalanine ureas. AB - Vitronectin receptor (alpha(V)beta(3)) antagonism has been implicated in a variety of disease states, like restenosis, osteoporosis and cancer. In this work, we present the development of a novel class of biphenyl vitronectin receptor antagonists. Identified from a focused combinatorial library based on para-bromo phenylalanine, these compounds show nanomolar affinity to the vitronectin receptor and display unprecedented SAR. Their binding mode can be rationalized by computational docking studies using the X-ray structure of alpha(V)beta(3). PMID- 12643915 TI - Synthesis and biological evaluation of 4-deacetoxy-1,7-dideoxy azetidine paclitaxel analogues. AB - Three novel 4-deacetoxy-1,7-dideoxy azetidine paclitaxel analogues were synthesized through a convenient route that employed hydroboration-amination and intramolecular S(N)2-type substitution reaction from a natural taxoid taxinine. All analogues have been tested for cytotoxicity against three human tumor cell lines. None of them showed remarkable cytotoxicity compared to paclitaxel against tested tumor cell lines. PMID- 12643916 TI - Visual detection of AMP and real-time monitoring of cyclic nucleotide phosphodiesterase (PDE) activity in neutral aqueous solution. Chemosensor-coupled assay of PDE and PDE inhibitors. AB - A phosphate chemosensor ensemble, [Zn(H-bpmp)(pyrocatechol violet)](+) is useful for a real-time assay of phosphodiesterases (PDEs) in a neutral aqueous solution. In addition, a simple and convenient screen and assay procedures for inhibitors of PDEs have been demonstrated using IBMX, a nonselective PDE inhibitor as an example. PMID- 12643917 TI - Regioselective hydrolysis of pentaacetyl catechin and epicatechin by porcine liver esterase. AB - 3,5,7,3',4'-Pentaacetyl catechin was selectively hydrolyzed to either 3,7,3',4' tetraacetyl catechin or 3-acetyl catechin depending upon the duration of hydrolysis. A similar result was also obtained in the epicatechin series. PMID- 12643918 TI - N-Alkoxysulfamide, N-hydroxysulfamide, and sulfamate analogues of methionyl and isoleucyl adenylates as inhibitors of methionyl-tRNA and isoleucyl-tRNA synthetases. AB - A series of sulfamate surrogates of methionyl and isoleucyl adenylate have been investigated as MetRS and IleRS inhibitors by modifications of the sulfamate linker and adenine moieties. The discovery of 2-iodo Ile-NHSO(2)-AMP (58) as a potent Escherichia coli IleRS inhibitor revealed that a significant hydrophobic interaction between the 2-substituent of Ile-NHSO(2)-AMP and the adenine binding site of IleRS provided its high potency to the enzyme. PMID- 12643919 TI - Discovery of 4'-[(imidazol-1-yl)methyl]biphenyl-2-sulfonamides as dual endothelin/angiotensin II receptor antagonists. AB - A series of 4'-[(imidazol-1-yl)methyl]biphenylsulfonamides has potent antagonist activity against both angiotensin II AT(1) and endothelin ET(A) receptors. Such dual-acting antagonists could have utility in the treatment of hypertension, heart failure, and other cardiovascular diseases in a broad patient population. Certain compounds in the present series are orally active in a rat model of angiotensin II-mediated hypertension. PMID- 12643921 TI - Protective effects of steroid saponins from Paris polyphylla var. yunnanensis on ethanol- or indomethacin-induced gastric mucosal lesions in rats: structural requirement for activity and mode of action. AB - The methanolic extract from the rhizomes of Paris polyphylla SM. var. yunnanensis (FR.) H-M. was found to potently inhibit ethanol-induced gastric lesions in rats. Through bioassay-guided separation, four known spirostanol-type steroid saponins, pennogenin 3-O-alpha-L-rhamnopyranosyl(1-->2)-[alpha-L-arabinofuranosyl(1-->4)] beta-D-glucopyranoside (1), pennogenin 3-O-alpha-L-rhamnopyranosyl(1-->4)-alpha-L rhamnopyranosyl(1-->4)-[alpha-L-rhamnopyranosyl(1-->2)]-beta-D-glucopyranoside (2), diosgenin 3-O-alpha-L-rhamnopyranosyl(1-->2)-[alpha-L-arabinofuranosyl(1- >4)]-beta-D-glucopyranoside (3), and diosgenin 3-O-alpha-L-rhamnopyranosyl(1-->4) alpha-L-rhamnopyranosyl(1-->4)-[alpha-L-rhamnopyranosyl(1-->2)]-beta-D glucopyranoside (4), and a new furostanol-type steroid saponin, parisaponin I (5), together with two known furostanol-type steroid saponins, trigofoenoside A (6) and protogracillin (7), were isolated from the active fraction. Compounds 1-4 (1.25-10 mg/kg, po) strongly inhibited gastric lesions induced by ethanol and indomethacin. With regard to structural requirement of steroid saponins, the 3-O glycoside moiety and spirostanol structure were found to be essential for the activity and the 17-hydroxyl group in the aglycon part enhanced the protective effects against ethanol-induced gastric lesions. The protective effects of 1 and 3 against ethanol-induced gastric lesions were attenuated by pretreatment with indomethacin and N-ethylmaleimide. Compounds 1 and 3 weakly inhibited acid secretions in pylorus-ligated rats. These findings suggested that endogenous prostaglandins and sulfhydryl compounds were involved in the protective activity. PMID- 12643920 TI - Synthesis and antibacterial study of 10, 15, 20-triphenyl-5-(4-hydroxy-3 (trimethylammonium)methyl)phenylporphyrin as models for combination of porphyrin and alkylating agent. AB - 10, 15, 20-Triphenyl-5-(4-hydroxy-3-(trimethylammonium)methyl)phenylporphyrin 4 and its zinc complex 5 have been synthesized and antibacterial activities have been studied for 4 and its derivative. Compound 4 showed stronger inhibition than that of 10, 15, 20-triphenyl-5-(4-hydroxy-3-(dimethylamine)methyl)phenylporphyrin (2) and 10, 15, 20-triphenyl-5-(4-methoxy-3 (trimethylammonium)methyl)phenylporphyrin (6). It is possible that antibacterial activity of compound 4 involved in photoinducing both o-quinone methide intermediate and singlet oxygen formation. PMID- 12643922 TI - Novel alpha-amino-acid phenolic ester derivatives with intravenous anaesthetic activity. AB - A novel series of alpha-amino-acid phenolic ester derivatives containing sulphide, sulphoxide, sulphone, ester and amide side chains were prepared and shown to display potent intravenous anaesthetic activity. PMID- 12643923 TI - Glycine alpha-ketoamides as HCV NS3 protease inhibitors. AB - Using a tetrapeptide-based alpha-ketoamide template, various amines and amino acids were incorporated to explore the prime side of the HCV NS3 protease catalytic site. Glycine carboxylic acid was found to be the most effective prime group. Further optimization yielded an inhibitor with IC(50) of 0.060 microM. PMID- 12643924 TI - Structure-based de novo design of non-nucleoside adenosine deaminase inhibitors. AB - We searched for non-nucleoside inhibitors of adenosine deaminase by rational structure-based de novo design and succeeded in the discovery of 1-(1-hydroxy-4 phenyl-2-butyl)imidazole-4-carboxamide (FR221647: K(i)=5.9 microM to human ADA) as a novel inhibitor with moderate activity and good pharmacokinetics compared with the known inhibitors pentostatin and EHNA. PMID- 12643925 TI - Conformationally-restricted analogues and partition coefficients of the 5-HT3 serotonin receptor ligands meta-chlorophenylbiguanide (mCPBG) and meta chlorophenylguanidine (mCPG). AB - The present investigation examined two features of arylbiguanide and arylguanidine 5-HT(3) ligands: conformation and partition coefficients. Several conformationally-constrained analogues of mCPBG (2) and mCPG (11; K(i)=32 nM) were prepared and of these only 2-amino-5-chloro-3,4-dihydroquinazoline (14; K(i)=34 nM) retained high affinity. The partition coefficient of compound 11 (LogP(app)=-0.64) was less than that of its corresponding arylbiguanide 2 (LogP(app)=-0.38). The quinazoline structure may represent a pharmacologically active conformation of these agents, and the arylbiguanides were found more lipid soluble than their arylguanidine counterparts at physiological pH. PMID- 12643926 TI - Preparation of potential inhibitors of the mur-pathway enzymes on solid support using an acetal linker. AB - Here, we report a novel strategy for the combinatorial or parallel solid-phase synthesis of potential inhibitors of the mur-pathway enzymes. The strategy involves the efficient use of p-alkoxybenzylidene acetal linker for reversible immobilization of sugar scaffolds to solid phase. This methodology was used to synthesize several glycopeptides on solid phase in good yields. PMID- 12643927 TI - Discovery of a potent and selective agonist of the prostaglandin EP4 receptor. AB - Analogues of PGE(2) wherein the hydroxycyclopentanone ring has been replaced by a lactam have been prepared and evaluated as ligands for the EP(4) receptor. An optimized compound (19a) shows high potency and agonist efficacy at the EP(4) receptor and is highly selective over the other seven known prostaglandin receptors. PMID- 12643928 TI - 1H-Pyrazolo[3,4-b]pyridine inhibitors of cyclin-dependent kinases. AB - 1H-Pyrazolo[3,4-b]pyridine 3 (SQ-67563) has been shown to be a potent, selective inhibitor of CDK1/CDK2 in vitro. In cells 3 acts as a cytotoxic agent with the ability to block cell cycle progression and/or induce apoptosis. The solid state structure of 3 bound to CDK2 shows 3 resides coincident with the ATP purine binding site and forms important H-bonding interactions with Leu83 on the protein backbone. PMID- 12643930 TI - Synthesis and Biological activity of kappa opioid receptor agonists. Part 2: preparation of 3-aryl-2-pyridone analogues generated by solution- and solid-phase parallel synthesis methods. AB - New analogues of the previously described 3-aryl pyridone KOR agonists have been synthesised by parallel synthetic methods, both in solution- and with solid-phase chemistry, making use of the well known and versatile Mitsunobu, Suzuki and Buchwald reactions. Opioid receptor binding data for the compounds produced is reported. PMID- 12643929 TI - Synthesis of imidazopyridines and purines as potent inhibitors of leukotriene A4 hydrolase. AB - The synthesis and biological evaluation of a series of heterocyclic analogues of the previously reported LTA(4) hydrolase inhibitor 1b are described. Imidazopyridine and purine analogues are specifically highlighted with several demonstrating excellent potency in our in vitro assays, as well as good oral activity in a mouse ex vivo assay. PMID- 12643931 TI - Bacterial siderophores: synthesis and biological activities of novel pyochelin analogues. AB - The synthesis and biological activities of four pyochelin analogues substituted in different parts of the molecule are reported: 5-NHBoc-pyochelin, 3"N-Boc pyochelin, 3"-nor-NH-pyochelin and neopyochelin II, the enantiomer of natural pyochelin. All these compounds complex iron(III) and transport it at different rates into the cells of Pseudomonas aeruginosa. PMID- 12643933 TI - Design and synthesis of potent, non-peptide inhibitors of HCV NS3 protease. AB - Starting from a hexapeptide boronic acid lead, 3-amino bicyclic pyrazinones as novel beta-sheet dipeptide mimetics have been designed and synthesized. Side chain manipulation of this scaffold generated a series of potent, nonpeptidic inhibitors of HCV NS3 protease. PMID- 12643932 TI - Novel ether derivatives of mannopeptimycin glycopeptide antibiotic. AB - Novel ether derivatives of mannopeptimycin glycopeptide were synthesized to probe their SAR. Many of these derivatives exhibited potent antibacterial activity against methicillin resistant and vancomycin resistant strains. These ether derivatives were prepared via reductive ring cleavage of acetals to give a mixture of 6-O, 4-O, 3-O, and 2-O-ether isomers. Both 6-O-ether and 4-O-ether showed significantly enhanced antibacterial activity over the parent and the isovalerate esters. PMID- 12643934 TI - Benzamide derivatives as blockers of Kv1.3 ion channel. AB - The voltage-gated potassium channel, Kv1.3, is present in human T-lymphocytes. Blockade of Kv1.3 results in T-cell depolarization, inhibition of T-cell activation, and attenuation of immune responses in vivo. A class of benzamide Kv1.3 channel inhibitors has been identified. The structure-activity relationship within this class of compounds in two functional assays, Rb_Kv and T-cell proliferation, is presented. In in vitro assays, trans isomers display moderate selectivity for binding to Kv1.3 over other Kv1.x channels present in human brain. PMID- 12643936 TI - A new and improved method for deglycosidation of glycopeptide antibiotics exemplified with vancomycin, ristocetin, and ramoplanin. AB - A general method for the deglycosidation of glycopeptide antibiotics has been developed. Treatment of vancomycin, ristocetin, and ramoplanin with anhydrous HF results in efficient cleavage of the sugars to provide the corresponding aglycons in high yield. PMID- 12643935 TI - Novel 5-cyclopropyl-1,4-benzodiazepin-2-ones as potent and selective I(Ks) blocking class III antiarrhythmic agents. AB - Novel 5-cyclopropyl-1,4-benzodiazepin-2-ones having various N-l substituents were identified as potent and selective blockers of the slowly activating cardiac delayed rectifier potassium current (I(Ks)). Compound 11 is the most potent I(Ks) channel blocker reported to date. PMID- 12643937 TI - Interfacial peptide inhibitors of HIV-1 integrase activity and dimerization. AB - Peptides derived from the interfacial region of dimeric HIV-1 integrase were evaluated as inhibitors of integrase's 3'-endonuclease activity. Three peptides were found to be moderately potent inhibitors with IC(50) values in the low micromolar range. The mode of inhibition was probed through protein crosslinking experiments. Active interfacial peptides were found to inhibit crosslinking of the dimeric form of integrase. Interfacial peptides that were poor inhibitors had no effect on integrase crosslinking. PMID- 12643938 TI - Synthesis and properties of 11-(3,5-di-tert-butyl-2 hydroxyphenylcarbamoyl)undecanoic acid, a new amphiphilic antioxidant. AB - Based on the membrane addressing concept, designing and synthesis of 11-(3,5-di tert-butyl-2-hydroxyphenylcarbamoyl)undecanoic acid have been carried out. Antioxidant properties of the prepared compound were investigated in comparison with its non-amphiphilic analogues. PMID- 12643939 TI - 2-(Anilinomethyl)imidazolines as alpha1 adrenergic receptor agonists: alpha1a subtype selective 2'-heteroaryl compounds. AB - The structure-activity relationship of 2'-pyrrole, pyrazole and triazole substituted 2-(anilinomethyl)imidazolines as alpha(1) adrenergic agonists was investigated. The size and orientation of substituents, as well as the position of the heteroatoms, were found to have a profound effect on the potency and selectivity of the molecules. Potent alpha(1A) subtype selective agonists have been identified. PMID- 12643940 TI - Structure-activity relationships studies of the anti-angiogenic activities of linomide. AB - The synthesis and anti-angiogenic activities of linomide and its analogues are reported. Three of the analogues are 3.3-69 times more potent than linomide at inhibiting blood vessel formation in the CAM angiogenesis assay. These compounds possessed considerable anti-proliferative activity against isolated HUVEC cells with no activity against epithelial-derived prostate tumor cells. PMID- 12643941 TI - N-Phenyl-N-purin-6-yl ureas: the design and synthesis of p38alpha MAP kinase inhibitors. AB - The design, synthesis and SAR of a series of 2,6,9-trisubstituted purine inhibitors of p38alpha kinase is reported. Synthetic routes were devised to allow for array synthesis in which all three points of diversity could be facilely explored. The binding of this novel series to p38alpha kinase, which was predicted to have several key interactions in common with SB-203580, was confirmed by X-ray crystallography of 19 (p38 IC(50)=82 nM). PMID- 12643942 TI - 3,4-Diaryl-5-hydroxyfuranones: highly selective inhibitors of cyclooxygenase-2 with aqueous solubility. AB - The introduction of a hydroxyl group into the 5-position of the diaryl furanone system provides highly selective inhibitors of cyclooxygenase-2. These molecules can be converted into their sodium salts which are water soluble, facilitating intravenous formulation. These salts show excellent potency in rat models of pain, fever and inflammation. PMID- 12643943 TI - Indanyl piperazines as melatonergic MT2 selective agents. AB - Optimization of a benzyl piperazine pharmacophore produced N-acyl-4-indanyl piperazines that bind with high affinity to melatonergic MT(2) receptors. (R)-4 (2,3-dihydro-6-methoxy-1H-inden-1-yl)-N-ethyl-1-piperazine-carboxamide fumarate (13) is a water soluble, selective MT(2) agonist, which produces advances in circadian phase in rats at doses of 1-56 mg/kg that are no different from those of melatonin at 1 mg/kg. Unlike melatonin, 13 produced only weak contractile effects in rat tail artery. PMID- 12643944 TI - Inhibitors of Abeta production: solid-phase synthesis and SAR of alpha hydroxycarbonyl derivatives. AB - Inhibitors of amyloid-beta (Abeta) protein production have been widely pursued as a potential treatment for Alzheimer's disease. Following the identification of a 5 microM screening hit, SAR was initiated using solid-phase synthetic techniques. Two series of alpha-hydroxy esters and ketones which are sub-micromolar inhibitors of Abeta production were identified. The most potent alpha hydroxyketone identified is approximately 30-fold more potent than the initial lead. PMID- 12643945 TI - Structure-activity relationships of some opiate glycosides. AB - A number of analogues of morphine-6-glucuronide 1 have been prepared and evaluated as potential analgesic agents by competitive mu-receptor binding assay and in vivo antinociceptive activity. The analogues show variation in the nature of the carbohydrate residue, the N-substituent, the O(3)-substituent and saturation of the 7,8-double bond compared to 1. In general, only the 6beta glucoside or beta-glucuronide carbohydrate residues showed potent agonism; other modified carbohydrates were less active or exhibited potential antagonism. Variations in N-substituent led to either reduced agonism (N-H) or potential antagonism [N-allyl, N-(cyclopropyl)methyl]; a polar N-substituent, carboxymethyl, failed to bind. Saturation of the 7,8-double bond led to increased agonism compared to the parent compound in all three examples studied. PMID- 12643946 TI - Azido-containing aryl beta-diketo acid HIV-1 integrase inhibitors. AB - Aryl beta-diketo acids (ADK) comprise a general class of potent HIV-1 integrase (IN) inhibitors, which can exhibit selective inhibition of strand transfer reactions in extracellular recombinant IN assays and provide potent antiviral effects in HIV-infected cells. Recent studies have shown that polycyclic aryl or aryl rings bearing aryl-containing substituents are components of potent members of this class. Reported herein is the first use of azido functionality as an aryl replacement in beta-diketo acid IN inhibitors. The ability of azido-containing inhibitors to exhibit potent inhibition of IN and antiviral protection in HIV infected cells, renders the azide group of potential value in the further development of ADK-based IN inhibitors. PMID- 12643947 TI - Race, Hispanic origin, and socioeconomic status in relation to motor vehicle occupant death rates and risk factors among adults. AB - Black and Hispanic adults travel less in motor vehicles than whites but may be at greater risk when they do travel. Passenger vehicle occupant deaths per 10 million trips among persons ages 25-64 were computed by race, Hispanic origin, gender, and socioeconomic status (SES) using 1995 data from the Fatality Analysis Reporting System (FARS) and Nationwide Personal Transportation Survey. Educational level was used as the indicator of SES. Blacks, particularly black men, were at increased risk of dying relative to whites when traveling in motor vehicles (rate ratio (RR) for black men=1.48; 95% confidence interval (CI)=1.42 1.54). Hispanic men, but not Hispanic women, also had elevated occupant death rates, but their risk was less than that of black men (RR=1.26; 95% CI=1.20 1.31). SES was the strongest determinant of occupant deaths per unit of travel; RRs among those who had not completed high school were 3.52 (95% CI=3.39-3.65) for men and 2.79 (95% CI=2.69-2.91) for women, respectively. Whites without high school degrees had the highest death rates per 10 million trips. After adjustment for SES, the elevated risk of occupant fatalities persisted among black men and women, but not among Hispanic men. Seat belt use and alcohol-impaired driving were examined among fatally injured passenger vehicle drivers. Among those with no education beyond high school, higher percentages were reported as having high blood alcohol concentrations (BACs) or having not used belts. Reported belt use rates were lower among black men and women, even after controlling for SES, whereas Hispanic men and women had belt use rates similar to those of whites. High BACs were more common among Hispanic men, which appeared largely to be an effect of SES because most Hispanic men killed in crashes had not completed high school, the education level with the highest percentage that drove while impaired by alcohol. More effective public health efforts are needed to reduce occupant deaths among persons of lower SES, blacks, and Hispanics, including measures to increase use of seat belts and reduce alcohol-impaired driving. PMID- 12643948 TI - Driving experience, crashes and traffic citations of teenage beginning drivers. AB - Teenagers were surveyed by telephone every 6 months from their freshman to senior high school years (N=911). Self-reported crash involvements and citations were examined for each teenager's first year of licensure and first 3500 miles driven. Based on survival analysis, the risk of a first crash during the first month of licensure (0.053) was substantially higher than during any of the next 11 months (mean risk per month: 0.025). The likelihood of a first citation during the first month of licensure (0.023) also was higher than during any of the subsequent 11 months (mean risk per month: 0.012). Similarly, when viewed as a function of cumulative miles driven, the risk of a first crash or citation was highest during the first 500 miles driven after licensure. Fewer parental restrictions (e.g. no nighttime curfew) and a lower grade point average (GPA) were associated with a higher crash risk. Male gender, a lower GPA and living in a rural area were associated with a higher citation rate. PMID- 12643949 TI - Driver risk factors for sleep-related crashes. AB - A population-based case-control study was carried out to examine driver risk factors for sleep-related motor vehicle crashes. Cases included 312 drivers involved in recent North Carolina crashes and identified on police reports as asleep at the time of the crash and 155 drivers identified as fatigued. Controls were 529 drivers also involved in recent crashes but not identified as asleep or fatigued, and 407 drivers not involved in recent crashes. All drivers were contacted for brief telephone interviews. Results showed that drivers in sleep related crashes were more likely to work multiple jobs, night shifts, or other unusual work schedules. They averaged fewer hours sleep per night, reported poorer quality sleep, were less likely to feel they got enough sleep, were sleepier during the day, drove more often late at night, and had more prior instances of drowsy driving. Compared to drivers in non-sleep-related crashes, they had been driving for longer times, been awake more hours, slept fewer hours the night before, and were more likely to have used soporific medications. Knowledge of specific risk factors for sleep-related crashes is an important first step in reducing the thousands of deaths and injuries each year in the US attributed to drowsy driving. PMID- 12643950 TI - Factors associated with fatal and injurious car crash among learner drivers in New South Wales, Australia. AB - This exploratory study investigated factors associated with car crash injury among learner drivers across difference ages by using data routinely collected by the NSW police. The results obtained indicated that some factors are commonly associated with car crash injury across nearly all ages. On the other hand, some others are more age specific. On the whole, female learner drivers were more at risk of being killed or injured as compared to males. The drivers of 16 years old had an increased risk of crash injury due to environmental factors, such as special road feature, and distraction outside the vehicle. The increased risk of crash injury for older drivers (> or =25 years) was associated with distractions from both inside and outside vehicle. Night time driving posed a special risk to learner drivers aged 20-24 years old, but not other age groups. Speeding was a common factor for the increased risk of crash injury across all age groups. The implication of the results and limitations of the study were discussed. PMID- 12643951 TI - Usability issues concerning child restraint system harness design. AB - A study was conducted to assess usability issues relating to child restraint system (CRS) harness design. Four convertible child restraint systems representing a wide variety of design features were used. Forty-two participants installed two child test dummies in both forward- and rear-facing configurations either inside or outside a test vehicle. Observer-scored checklists determined the degree to which each harness was installed correctly. Participant-scored questionnaires evaluated the 'ease-of-use' of various design features. While the percentage of correct installations exceeded 83% for all designs when installed in the forward-facing configuration, in the rear-facing position (that intended for children under 9-10 kg), there was a significant (between 65 and 89%) percentage of incorrect installations for all models. This finding is of particular interest and may be indicative of a more generalized problem with 'convertible' CRS designs when they are used in the rear-facing configuration. Furthermore, while certain design features were perceived by users as providing significantly better protection in the event of a collision, these also tended to be the features that were misused most often. The benefits and costs of various design features are discussed, and a method to test harness design usability is presented. PMID- 12643952 TI - Evaluating the reliability of automated collision notification systems. AB - The use of an automated collision notification (ACN) device in vehicles can greatly reduce the time between crash occurrence and notification of emergency medical services (EMSs). Most ACN devices rely on cellular technology to report important crash information to the proper authorities. The objective of this study was to examine the ability of the existing western New York cellular analog system to support ACN systems. The first task was to develop a model predicting the probability of successfully completing an emergency ACN call at attenuated levels of received signal strength indicator (RSSI), a measurement of the bond between cell phone and tower. Then, empirical estimates were made of the time necessary for call completion at given levels of the RSSI. The RSSI is sampled at locations throughout Erie County, New York, and this information is used to determine the probability of successful call completion for different locations within the county. This model was then applied to historic data for selected past crashes. Finally, the findings were compared with real-world crash data obtained from the ACN Field Operational Test program, where 750 ACN devices were installed in cars and their performance examined over time. An interpolated map of the sampled RSSI values suggests that cellular coverage in Erie County is adequate to support the automated collision network technology. The models and techniques described here are applicable to other areas and regions of the country. PMID- 12643953 TI - The paradox of driving speed: two adverse effects on highway accident rate. AB - Whereas speeding is known to be a substantial risk factor in driving, there is no unequivocal evidence that accident rate on limited-access motor highways is considerably affected by average speed or by speed limits meant to regulate it. It is suggested here that the seeming puzzle actually may have a straightforward explanation: accident-prone interactions (APIs) between cars occur when they pass each other--mostly moving in the same directions or in opposite ones. Such interactions are shown here to happen more frequently, the lower average speed is. To the extent that high speed limits contribute to increase in average speed, they serve to reduce the number of such interactions, thereby to moderate at least part of the negative effect of speed on the driver's ability to avoid an impending accident. PMID- 12643954 TI - Factors influencing injury severity of motor vehicle-crossing pedestrian crashes in rural Connecticut. AB - The ordered probit model was used to evaluate the effect of roadway and area type features on injury severity of pedestrian crashes in rural Connecticut. Injury severity was coded on the KABCO scale and crashes were limited to those in which the pedestrians were attempting to cross two-lane highways that were controlled by neither stop signs nor traffic signals. Variables that significantly influenced pedestrian injury severity were clear roadway width (the distance across the road including lane widths and shoulders, but excluding the area occupied by on-street parking), vehicle type, driver alcohol involvement, pedestrian age 65 years or older, and pedestrian alcohol involvement. Seven area types were identified: downtown, compact residential, village, downtown fringe, medium-density commercial, low-density commercial, and low-density residential. Two groups of these area types were found to experience significantly different injury severities. Downtown, compact residential, and medium- and low-density commercial areas generally experienced lower pedestrian injury severity than village, downtown fringe, and low-density residential areas. PMID- 12643955 TI - Monotony of road environment and driver fatigue: a simulator study. AB - Studies have shown that drowsiness and hypovigilance frequently occur during highway driving and that they may have serious implications in terms of accident causation. This paper focuses on the task induced factors that are involved in the development of these phenomena. A driving simulator study was conducted in order to evaluate the impact of the monotony of roadside visual stimulation using a steering wheel movement (SWM) analysis procedure. Fifty-six male subjects each drove during two different 40-min periods. In one case, roadside visual stimuli were essentially repetitive and monotonous, while in the other one, the environment contained disparate visual elements aiming to disrupt monotony without changing road geometry. Subject's driving performance was compared across these conditions in order to determine whether disruptions of monotony can have a positive effect and help alleviate driver fatigue. Results reveal an early time on-task effect on driving performance for both driving periods and more frequent large SWM when driving in the more monotonous road environment, which implies greater fatigue and vigilance decrements. Implications in terms of environmental countermeasures for driver fatigue are discussed. PMID- 12643956 TI - The efficacy of an educational intervention in promoting self-regulation among high-risk older drivers. AB - Visual processing impairment increases crash risk among older drivers. Many older drivers meet the legal requirements for licensing despite having vision impairments that elevate crash risk. In this study, 365 older drivers who were licensed, visually-impaired, and crash-involved in the prior year were randomly assigned to an intervention group or usual-eye-care control group to evaluate the efficacy of an educational intervention that promoted the performance of self regulatory practices. The educational curriculum was designed to change self perceptions about vision impairment and how it can impact driver safety and to promote the avoidance of challenging driving situations through self-regulation, leading to reductions in driving exposure. Analyses compared the intervention and control groups at pre-test and 6 months post-test with respect to self-reported perceptions about vision and driving practices. At post-test, drivers who had received the educational intervention were more likely to acknowledge that the quality of their eyesight was less than excellent, report a higher frequency of avoiding challenging driving situations (e.g. left-turns) and report performing more self-regulatory practices (e.g. three right-turns) as compared to controls. Additionally, drivers in the educational intervention group reported significantly fewer days, fewer places and fewer trips made per week as compared to those not receiving the educational intervention. These findings imply that visually-impaired older drivers at higher risk for crash involvement may benefit from educational interventions by reducing their driving exposure and increasing their avoidance of visually challenging driving situations. A critical future step in this research program will be to examine whether this educational intervention has an impact on the safety of these high-risk older drivers by reducing their crash involvement in the years following the educational intervention. PMID- 12643957 TI - Tiredness! a natural explanation to The Grand Rapid "DIP". AB - The present analysis deals with the presumed improved driving skill of car drivers with low blood alcohol concentrations (BACs) compared to the driving skill of sober drivers. Several roadside surveys indicate such a controversial possibility, which is illustrated graphically in "The Grand Rapid Study" (1964) as a relative accident risk curve for driving while intoxicated (DWI). A curve which shows a "DIP" below a baseline accident risk of one for sober drivers in the interval between BACs of 0.01 and 0.04 g/l. Since then, various attempts have been made to show this "DIP" in the otherwise "exponentially" raising curve to be an artificial distortion based on disproportionate demographic subgroups. It is, however, our thesis that the observations of "The Grand Rapid Study" and other roadside surveys are valid, because the presumed mono-causal traffic accident curve hides "tiredness" as an additional human risk factor. This makes the first part of the night-time relative accident risk curve for impaired drivers artificially augmented by "tired" drivers with insignificant amounts of alcohol in the blood and therefore incorrectly accused of accidents due to DWI. The accident risk curve for higher BACs raises similarly abrupt due to drivers impaired by a combined effect of alcohol and "tiredness". Moreover, to imply an accident risk of one for all sober drivers independent of the time of the day is debatable as an increased accident risk is present in the late night-time hours due to "tired" drivers. This increase in "tired" sober accident drivers suppresses the accident risk curve for DWI with BACs of 0.01-0.04 g/l below the fixed baseline of one for the background population. PMID- 12643958 TI - Trends in fatal crashes involving female drivers, 1975-1998. AB - Since the mid-1980s there has been concern about the growing number of female drivers in the US involved in fatal motor vehicle crashes, and similar trends have been noted in other parts of the world. The present study examined whether this trend has continued into the 1990s and the reasons for it. Fatal crash data were obtained from the Fatality Analysis Reporting System (FARS), mileage data from the National Personal Transportation Survey, and licensure data from the Federal Highway Administration. Many more women were licensed to drive in 1998 than in 1975, and on average they drove more miles. When changes in total annual mileage were taken into account, per-mile crash rates decreased similarly for men and women (about 40%). An examination of the characteristics of their fatal crashes revealed that male and female drivers have seen similar reductions in single-vehicle, nighttime, and alcohol-related crashes. However, men continue to be involved more often in these types of crashes. PMID- 12643959 TI - Upper extremity interaction with a deploying side airbag: a characterization of elbow joint loading. AB - Computer simulations, dummy experiments with a new enhanced upper extremity and small female cadaver experiments were used to analyze the small female upper extremity response under side airbag loading. After establishing a worst case initial position, three tests were performed with the fifth percentile female hybrid III anthropometric test dummy and six experiments with small female cadaver subjects. A new fifth percentile female enhanced upper extremity was developed for the dummy experiments that included a two-axis wrist load cell in addition to the existing six-axis load cells in both the forearm and humerus. Forearm pronation was also included in the new dummy upper extremity to increase the biofidelity of the interaction with the handgrip. Instrumentation for both the cadaver and dummy tests included accelerometers and MHD angular rate sensors on the forearm, humerus, upper and lower spine. In order to quantify the applied loads to the cadaver hand and wrist from the door mounted handgrip, the handgrip was mounted to the door through a five-axis load cell and instrumented with accelerometers for inertial compensation. All six of the cadaver tests resulted in upper extremity injuries including comminuted mid-shaft humerus fractures, osteochondral fractures of the elbow joint surfaces, a transverse fracture of the distal radius and an osteochondral fracture of the lunate carpal bone. The results from the 6 cadaver tests presented in this study were combined with the results from 12 previous cadaver tests. A multivariate logistic regression analysis was performed to investigate the correlation between observed injuries and measured occupant response. Using inertially compensated force measurements from the dummy mid-shaft forearm load cell, the linear combination of elbow axial force and shear force was significantly (P=0.05) correlated to the observed elbow injuries. PMID- 12643960 TI - A comparison of headway and time to collision as safety indicators. AB - The two safety indicators "headway" and "time to collision (TTC)" are discussed and compared with respect to their usefulness in determining the safety of different traffic situations, like different locations in a junction. Over a 6 day-period traffic flow measures were taken in a four-way junction with stop signs on the minor road. It was found that for vehicles in a car following situation headway and TTC are independent of each other. The percentage of small headways is relatively constant across different locations in the junction, while the percentage of small TTC values varies between different locations. It is recommended to use headway for enforcement purposes, because small headways generate potentially dangerous situations. TTC, on the other hand, should be used when a certain traffic environment is to be evaluated in terms of safety, because it indicates the actual occurrences of dangerous situations. PMID- 12643961 TI - Modelling risk factors for injuries from dog bites in Greece: a case-only design and analysis. AB - We conducted a study using a newly developed dataset based on Emergency Departments records of a network of hospitals from Greece on injuries from dog bites. Our goal is three-fold: (a) to investigate if surrogate factors of leisure time are associated with increased risk of injury from bites; (b) to address recently reported contradictory results on putative association of lunar periods and injuries from dog bites; and (c) to offer a general methodology for addressing similar case-only designs with combined factors of which some can exhibit cyclical patterns. To address these goals, we used a case-only design of our dataset, and conducted an analysis where we controlled simultaneously for weekday/weekend effects, season of year (winter, spring/fall, summer), and lunar periods, because any one of these factors can contribute to the degree of exposure to injuries from dog bites. We found that increased risk of injury from bites was associated with weekends versus weekdays (RR=1.19, 95% CI: 1.10-1.29), summer versus winter (RR=1.24, 95% CI: 1.11-1.39), and fall or spring versus winter (RR=1.31, 95% CI: 1.19-1.45). The results support the hypothesis that longer leisure time at these levels of factors does increase the risk of having a bite injury. Moreover, after controlling for these factors, risk of bite injury was not associated with moon periods, thereby also helping settle a longstanding argument. PMID- 12643963 TI - Reality monitoring and metacognitive beliefs related to cognitive confidence in obsessive-compulsive disorder. AB - The present study investigated general reality monitoring ability, and selective reality monitoring ability for anxiety relevant actions in a group of individuals with obsessive-compulsive disorder (OCD) and a group of non-anxious controls. In addition, reality monitoring confidence was assessed, as well as specific meta cognitive beliefs related to cognitive confidence (by means of the Meta Cognitions Questionnaire (MCQ)). No differences were found between both groups in actual reality monitoring ability. Unlike previous studies, the reality monitoring task included actions that were related to the individual concerns of the OCD patients and were ideographically selected. Nevertheless, no differential reality monitoring effect was observed for the anxiety relevant stimuli. Data from the MCQ, however, revealed that OCD patients had less overall confidence in their memory for actions and their reality monitoring ability. Analysis of the confidence ratings of the reality monitoring task showed that this reduced confidence was restricted to the neutral actions. No differences were observed for patients that reported low or high frequencies of checking behaviour. The whole of these data do not support memory deficit models of OCD, but are in line with recent emphasis on the importance of memory confidence and other meta cognitive beliefs in OCD. PMID- 12643964 TI - Emotional avoidance: an experimental test of individual differences and response suppression using biological challenge. AB - The present study examined the affective consequences of response inhibition during a state of anxiety-related physical stress. Forty-eight non-clinical participants were selected on the basis of pre-experimental differences in emotional avoidance (high versus low) and subjected to four inhalations of 20% carbon dioxide-enriched air. Half of the participants were instructed to inhibit the challenge-induced aversive emotional state, whereas the other half was instructed to simply observe their emotional response. Participants high in emotional avoidance compared to those low in emotional avoidance responded with greater levels of anxiety and affective distress but not physiological arousal. Individuals high in emotional avoidance also reported greater levels of anxiety relative to the low emotional avoidance group when suppressing compared to observing bodily sensations. These findings are discussed in terms of the significance of emotional avoidance processes during physical stress, with implications for better understanding the nature of panic disorder. PMID- 12643965 TI - The impact of verbal feedback about blushing on social discomfort and facial blood flow during embarrassing tasks. AB - To investigate whether verbal feedback about blushing influences subsequent social discomfort or blushing itself, changes in facial blood flow and ratings of blushing and embarrassment were investigated in high (N=28) and low scorers (N=28) on the Blushing Propensity scale while singing and reading aloud, and while listening to audiotapes of their performance. After each task half of the participants were told that they had blushed, and the rest were told that they had not blushed. Blood flow increased progressively in participants with high blushing propensity scores who were given "blushing" feedback, but not in the other participants. This finding suggests that expecting to blush may become a self-fulfilling prophecy. Verbal feedback about blushing strongly influenced subsequent social discomfort, and mimicked the effects of blushing propensity on ratings of embarrassment and blushing intensity. In sum, the findings support the view that preconceptions about blushing propensity are shaped by past learning experiences, that concern about blushing is a major source of discomfort in embarrassing situations, and that these concerns are often unrelated to the actual intensity of blushing. PMID- 12643966 TI - Quantitative criteria for insomnia. AB - Formal diagnostic systems (DSM-IV, ICSD, and ICD-10) do not provide adequate quantitative criteria to diagnose insomnia. This may not present a serious problem in clinical settings where extensive interviews determine the need for clinical management. However, lack of standard criteria introduce disruptive variability into the insomnia research domain. The present study reviewed two decades of psychology clinical trials for insomnia to determine common practice with regard to frequency, severity, and duration criteria for insomnia. Modal patterns established frequency (> or =3 nights a week) and duration (> or =6 months) standard criteria. We then applied four versions of severity criteria to a random sample and used sensitivity-specificity analyses to identify the most valid criterion. We found that severity of sleep onset latency or wake time after sleep onset of: (a) > or =31 min; (b) occurring > or =3 nights a week; (c) for > or =6 months are the most defensible quantitative criteria for insomnia. PMID- 12643967 TI - Cognitive behaviour therapy with adults with intractable epilepsy and psychiatric co-morbidity: preliminary observations on changes in psychological state and seizure frequency. AB - Cognitive behaviour therapy (CBT) was undertaken with six adults with chronic, poorly controlled seizures and co-existing psychiatric and/or psychosocial difficulties. During 12 sessions of CBT from an experienced CBT Nurse Specialist, treatment focused concurrently on epilepsy-related problems, associated psychopathology and on the development of psychological strategies to reduce seizure occurrence. At the end of treatment participants rated their initial epilepsy-related problem as having less impact on their daily lives and at one month follow-up reported less deleterious impact on everyday life in terms of their psychological difficulties. In addition participants demonstrated significant improvements in terms of their self-rated work and social adjustment, and in their decreased use of escape-avoidance coping strategies. These positive findings occurred despite the absence of a significant decrease in seizure frequency. Issues raised by the complexity and severity, both of these patients' psychological/psychosocial difficulties and their epilepsy, are discussed in relation to the optimal length of treatment that may be required when adopting a CBT model in this patient group. PMID- 12643969 TI - 'Minor GAD': characteristics of subsyndromal GAD in older adults. AB - Subsyndromal emotional symptoms are common in older adults and are associated with increased disability, health care utilization, and risk for developing psychiatric disorders. The purpose of this study was to examine subsyndromal generalized anxiety disorder (GAD) in older adults. Participants included 30 older adults with diagnosable GAD, 19 with subsyndromal anxiety symptoms [minor GAD; (MGAD)], and 21 normal control volunteers (NC). Participants were assessed using the Anxiety Disorders Interview Schedule for DSM-IV and completed self report measures of anxiety, worry, depression, and life satisfaction. Excessive worry on more days than not, difficulty controlling worry, and clinically significant distress or impairment were the diagnostic criteria endorsed by MGAD participants least often. Therefore, these criteria may be useful in distinguishing between GAD and subsyndromal GAD. Self-reported anxiety and worry also systematically differed across groups in the expected directions, with a discriminant analysis yielding good classification of the GAD and NC groups based on these measures. Categorization of MGAD participants generally was poor, with most misclassified as GAD patients. Clinical implications of these findings are discussed. PMID- 12643970 TI - Cognitive behaviour therapy of acute stress disorder: a four-year follow-up. AB - The aim of this study was to index the long-term benefits of early provision of cognitive behavior therapy to trauma survivors with acute stress disorder. Civilian trauma survivors (n = 80) with acute stress disorder were randomly allocated to either cognitive behavior therapy (CBT) or supportive counseling (SC) - 69 completed treatment, and 41 were assessed four years post-treatment for post-traumatic stress disorder (PTSD) with the Clinician Administered PTSD Scale. Two CBT patients (8%) and four SC patients (25%) met PTSD criteria at four-year follow-up. Patients who received CBT reported less intense PTSD symptoms, and particularly less frequent and less avoidance symptoms, than patients who received SC. These findings suggest that early provision of CBT in the initial month after trauma has long-term benefits for people who are at risk of developing PTSD. PMID- 12643968 TI - Phobic anxiety in 11 nations. Part I: Dimensional constancy of the five-factor model. AB - The Fear Survey Schedule-III (FSS-III) was administered to a total of 5491 students in Australia, East Germany, Great Britain, Greece, Guatemala, Hungary, Italy, Japan, Spain, Sweden, and Venezuela, and submitted to the multiple group method of confirmatory analysis (MGM) in order to determine the cross-national dimensional constancy of the five-factor model of self-assessed fears originally established in Dutch, British, and Canadian samples. The model comprises fears of bodily injury-illness-death, agoraphobic fears, social fears, fears of sexual and aggressive scenes, and harmless animals fears. Close correspondence between the factors was demonstrated across national samples. In each country, the corresponding scales were internally consistent, were intercorrelated at magnitudes comparable to those yielded in the original samples, and yielded (in 93% of the total number of 55 comparisons) sex differences in line with the usual finding (higher scores for females). In each country, the relatively largest sex differences were obtained on harmless animals fears. The organization of self assessed fears is sufficiently similar across nations to warrant the use of the same weight matrix (scoring key) for the FSS-III in the different countries and to make cross-national comparisons feasible. This opens the way to further studies that attempt to predict (on an a priori basis) cross-national variations in fear levels with dimensions of national cultures. PMID- 12643971 TI - Inflated responsibility and the use of stop rules for catastrophic worrying. AB - The present paper reports the results of two experiments designed to test some predictions from a mood-as-input explanation of catastrophic worrying (). In particular, these experiments attempted to identify whether worriers possess characteristics that would contribute to the use of relatively strict 'as many as can' closure rules for catastrophising. Experiment 1 demonstrated that high worriers begin a catastrophising task with higher self-reported levels of responsibility towards fully considering all issues involved, than low worriers. Experiment 2 suggested that inflated responsibility has a causal effect on perseveration at the catastrophising task (rather than being a simple non-causal by-product of excessive worrying), and that inflated responsibility exacerbates catastrophising only in conjunction with negative mood. This suggests a relatively complex relationship between responsibility and mood, where there are mood conditions in which high responsibility does not generate greater persistence than low responsibility. These findings are consistent with predictions from a mood-as-input account of catastrophic worrying, and provide evidence for a putative mechanism that mediates the influence of variables such as inflated responsibility on perseveration. PMID- 12643975 TI - The generation of free radicals by nitric oxide synthase. AB - Nitric oxide synthase (NOS) is an example of a family of heme-containing monooxygenases that, under the restricted control of a specific substrate, can generate free radicals. While the generation of nitric oxide (NO*) depends solely on the binding of L-arginine, NOS produces superoxide (O(2)*(-)) and hydrogen peroxide (H(2)O(2)) when the concentration of the substrate is low. Not surprisingly, effort has been put forth to understand the pathway by which NOS generates NO*, O(2)*(-) and H(2)O(2), including the role of substrate binding in determining the pathways by which free radicals are generated. By binding within the distal heme pocket near the sixth coordination position of the NOS heme iron, L-arginine alters the coordination properties of the heme iron that promotes formation of the perferryl complex NOS-[Fe(5+)=O](3+). This reactive iron intermediate has been shown to abstract a hydrogen atom from a carbon alpha to a heteroatom and generate carbon-centered free radicals. The ability of NOS to produce free radicals during enzymic cycling demonstrates that NOS-[Fe(5+)=O](3+) behaves like an analogous iron-oxo complex of cytochrome P-450 during aliphatic hydroxylation. The present review discusses the mechanism(s) by which NOS generates secondary free radicals that may initiate pathological events, along with the cell signaling properties of NO*, O(2)*(-) and H(2)O(2). PMID- 12643976 TI - Effect of bursal anti-steroidogenic peptide and immunoglobulin G on neonatal chicken B-lymphocyte proliferation. AB - In attempts to identify antibodies for Bursal Anti-Steroidogenic Peptide (BASP), rabbit serum was observed to reduce phorbol ester-stimulated chicken B-lymphocyte proliferation comparable to BASP. These experiments investigated the effects of IgG on B-lymphocyte proliferation. In Experiment 1, 3% rabbit serum decreased B lymphocyte proliferation. In Experiment 2, 2 mg/ml of intact rabbit IgG or 0.65 mg/ml of IgG papain digest products, Fab and Fc, decreased B-lymphocyte proliferation. The combination of BASP and either Fab or Fc was observed to have at least an additive anti-proliferative effect. In Experiment 3, 0.01 mg/ml of either rabbit or chicken IgG, or 1.0 mg/ml of rabbit or 0.01 mg/ml of chicken Fab, Fc, and the pepsin digestion product F(ab')(2) was observed to have an anti proliferative effect. No combined effects of BASP and IgG or IgG digest products were observed for this experiment. In Experiment 4, 12 mg/ml of chicken egg yolk IgG or 1.2 mg/ml Fab was found to suppress B-lymphocyte proliferation. Additionally, an additive effect of 12 mg/ml of IgG with BASP was again observed. The present studies suggest that IgG and its digestion products reduce phorbol stimulated B-lymphocyte proliferation in vitro and combined treatment with IgG and BASP may have at least an additive anti-proliferative effect on B-lymphocyte proliferation. PMID- 12643977 TI - In vitro proliferation of blood, spleen and thymus leukocytes from the turtle Mauremys caspica. AB - Cell-mediated immune responses is commonly evaluated by cell proliferation assays. Mitogens are known to induce a vigorous proliferative response in lymphoid cells from mammals but relatively fewer studies have investigated mitogen-mediated lymphoproliferation in non-mammalian animals. In the present work, we incubated spleen, thymus and blood leukocytes with phytohaemagglutinin (PHA), concanavalin A (Con A), lipopolysaccharide (LPS) and pokeweed mitogen (PWM), by different times of incubation (96 and 120 h) and at different concentrations. Our results show that the optimal mitogen concentrations inducing proliferation on leukocytes from Mauremys caspica were 20 microg/ml PHA, 1 microg/ml Con A, 12.5 microg/ml LPS and 1/150 dilution PWM. The optimal time of incubation was dependent on the type of leukocytes (peripheral blood leukocytes, splenic leukocytes or thymic cells) and the mitogen utilized. PMID- 12643978 TI - Metals in squid, Loligo forbesi, adults, eggs and hatchlings. No evidence for a role for Cu- or Zn-metallothionein. AB - An adult squid Loligo forbesi had the following metals in its liver/digestive gland: Mn, Fe, Ni, Zn, Cu, As, Cd, Ba and Pb in the range of 1-110 ppm wet wt. Adult mantle muscle, adult eyes, eggs and hatchlings contained a lesser number of these metals at concentrations above 1 ppm. Chromatographic analysis of non-heat treated cytosols (in the presence of 5 mM 2-mercaptoethanol) gave no evidence for the presence of copper- or zinc-containing fractions with the molecular weights of mollusc metallothioneins in any of the above tissues. Copper and Zn were bound to either the particulate fraction or to very low molecular weight species. PMID- 12643979 TI - Aluminum-induced oxidative stress in rat brain: response to combined administration of citric acid and HEDTA. AB - Aluminum, a known neurotoxic substance, has been suggested as a contributing factor in the pathogenesis of Alzheimer's disease. Therapeutic efficacy of combined administration of citric acid (CA) and N-(2-hydroxyethyl) ethylenediaminetriacetic acid (HEDTA) was evaluated in decreasing blood and brain aluminum concentration and parameters indicative of hematological disorders and brain oxidative stress. Adult male wistar rats were exposed to drinking water containing 0.2% aluminum nitrate for 8 months and treated once daily for 5 consecutive days with CA (50 mg/kg, orally) or HEDTA (50 mg/kg, intraperitoneally) either individually or in combination. Aluminum exposure significantly inhibited blood delta-aminolevulinic acid dehydratase while increased zinc protoporphyrin confirming changed heme biosynthesis. Significant decrease in the level of glutathione S-transferase in various brain regions and an increase in whole brain thiobarbituric acid reactive substance, and oxidized glutathione (GSSG) levels were also observed. Glutathione peroxidase activity showed a significant increase in cerebellum of aluminum exposed rats. Most of the above parameters responded moderately to the individual treatment with CA and HEDTA, but significantly reduced blood and brain aluminum burden. However, more pronounced beneficial effects on some of the above described parameters were observed when CA and HEDTA were administered concomitantly. Blood and brain aluminum concentration however, showed no further decline on combined treatment over the individual effect with HEDTA or CA. We conclude that in order to achieve an optimum effect of chelation, combined administration of CA and HEDTA might be preferred. However, further work is needed before a final recommendation could be made. PMID- 12643980 TI - Effect of oxidants on small intestinal brush border membranes and colonic apical membranes--a comparative study. AB - This study compares composition of the rat small intestinal brush border membranes (BBM) and colonic apical membranes (CAM) and their susceptibility to in vitro exposure to various oxidants. Differences were observed between BBM and CAM in their lipid composition, sugar content, alkaline phosphatase (ALP) activity and cholesterol/phospholipid ratio. BBM and CAM were exposed to superoxide generated by xanthine+xanthine oxidase (X-XO) or peroxides such as tertiary butyl hydroperoxide (tBuOOH) and hydrogen peroxide (H(2)O(2)) and alterations in ALP activity, peroxidation parameters and membrane lipids were analyzed. Exposure of BBM and CAM to superoxide resulted in decrease in ALP activity and increase in peroxidation parameters such as protein carbonyl content, malondialdehyde and conjugated diene. Superoxide exposure also resulted in lipid alterations specifically in certain phospholipids. These alterations were prevented either by superoxide dismutase or by allopurinol. Peroxides did not have any significant effect. These results suggest that both BBM and CAM are susceptible to superoxide, which can bring about peroxidation and degradation of membrane lipids specifically, certain phospholipids. PMID- 12643981 TI - Variation in fibroblast growth factor response and heparan sulfate proteoglycan production in satellite cell populations. AB - Variation in the responsiveness of myogenic satellite cell subpopulations to mitogenic stimuli was examined in cells isolated from the turkey pectoralis major muscle. Faster growing clonal cell populations were more responsive to fibroblast growth factor (FGF-2) and expressed greater levels of FGF-2 and FGF receptor-1 at the onset of proliferation than did slower growing cells. Faster growing clones also expressed higher levels of heparan sulfate proteoglycans (HSPG), especially during differentiation, than did slower growing clones. HSPG, which is important in FGF receptor signaling, increased during proliferation of all clones tested and decreased in all but one of the clones during differentiation. Slower growing clones increased their expression of FGF receptor-1 through proliferation and differentiation. However, expression of the receptor in faster growing clones decreased during differentiation. The FGF receptors-2 and -3 were not detected on turkey satellite cells or myotubes by reverse transcriptase-polymerase chain reaction methodology. These results demonstrate that there is heterogeneity in the properties and responsiveness of satellite cell populations derived from single muscles. Satellite cells that differ in proliferation rates differ in responsiveness to FGF-2, and in the expression of FGF-2, FGF receptor-1, and HSPG. PMID- 12643982 TI - p-Nitrophenol and glutathione response in medaka (Oryzias latipes) exposed to MX, a drinking water carcinogen. AB - When chlorine is introduced into public drinking water for disinfection, it can react with organic compounds in surface waters to form toxic by-products such as 3-chloro-4-(dichloromethyl)-5-hydroxy-2[5H]-furanone (MX). We investigated the effect of exposure to MX on cytochrome P450 2E1 (CYP2E1)-like activity and total glutathione (GSH) in the liver of the small fish model, medaka (Oryzias latipes). The multi-site carcinogen methylazoxymethanol acetate (MAMAc) was the positive control compound. Both medaka liver microsome preparations and S-9 fractions catalyzed the hydroxylation of p-nitrophenol (PNP), suggesting CYP2E1-like activity in the medaka. Male medaka exposed for 96 h to the CYP2E1 inducers ethanol and acetone under fasted conditions showed significant increases in PNP hydroxylation activity. Furthermore, total reduced hepatic GSH was reduced in fish fasted for 96 h, indicating that normal feeding is a factor in maintaining xenobiotic defenses. Exposure to MX and MAMAc induced significant increases in hepatic CYP2E1-like activity, however MX exposure did not alter hepatic GSH levels. These data strengthen the role of the medaka as a suitable species for examining cytochrome P450 and GSH detoxification processes and the role these systems play in chemical carcinogenesis. PMID- 12643983 TI - Effects of exposure to 17alpha-ethinylestradiol during early development on sexual differentiation and induction of vitellogenin in zebrafish (Danio rerio). AB - To determine the critical stage of zebrafish development where exposure to xenoestrogens can affect sex ratio and vitellogenin induction, zebrafish (Danio rerio) were exposed to 17alpha-ethinylestradiol (actual concentration 15.4+/-1.4 ng EE2/l) during early development: from fertilisation to hatch; hatch to 10 days post hatch (dph); 10-20 dph; 20-30 dph; 20-40 dph; 20-60 dph; fertilisation to 25 dph; or hatch to 60 dph. Vitellogenin was measured in whole body homogenate 30 dph by ELISA and sex ratio was determined 60 dph by histological examination of the gonads. All exposure periods significantly induced vitellogenin synthesis and affected the sex differentiation leading to development of ovo-testis or complete feminisation of the exposed fish depending on exposure period. Complete sex reversal was obtained in groups exposed from 20 to 60 dph and hatch to 60 dph. The half-life for degradation of vitellogenin was calculated. Juvenile zebrafish were exposed to 15.4+/-1.4 ng EE2/l (actual concentration) from fertilisation to 25 dph and transferred to clean water, after which weekly measurements of vitellogenin concentration in whole body homogenate were performed until day 46 post hatch. The half-life of vitellogenin was 2.4 days. PMID- 12643984 TI - Effects of small concentrations of mercury on the contractile activity of the rat ventricular myocardium. AB - Personal exposure to mercury vapor and the release of mercury from or during removal of amalgam dental fillings increases its blood and plasma concentration. However, it is not known if these very small amounts affect cardiac function. The effects of continuous exposure to 5 and 20 nM of HgCl(2) on the cardiac contractility were investigated in isometric and tetanic contractions of right ventricular strips and in Langendorff perfused rat hearts. The continuous exposure for 2 h produced a small but significant reduction of the isometric twitch force and time to peak tension shortened. Relative post-rest potentiation was not affected by this concentration of HgCl(2) suggesting a lack of action of the metal on the sarcoplasmic reticulum activity. Tetanic tension, in contrast to twitch force, was intensively reduced suggesting an important depressant action on the activity of contractile proteins. In perfused hearts beating spontaneously, isovolumic systolic pressure reduced progressively and the diastolic pressure increased. Although occurring heart rate reduction, it was similar for both controls and mercury treated hearts. Also, time dependent changes in coronary perfusion pressure were similar to controls. Results suggested that cardiac effects may be observed after continuous exposure to very small concentrations of mercury, probably as a result of the cell capacity to concentrate mercury. These results also indicate that continuous professional exposure to mercury followed by its absorption might have toxicological consequences affecting cardiac function, and being considered hazardous. PMID- 12643985 TI - Circadian variations of plasma renin activity (PRA), aldosterone and electrolyte concentrations in plasma in pregnant and non-pregnant goats. AB - The aim of this study was to estimate and analyse circadian variations of the renin-angiotensin-aldosterone system (RAA) activity in blood of goats and the influence of late pregnancy on the circadian variations of RAA system. The study was carried out on a group of 17 non-pregnant and 9 pregnant goats. The animals were kept in uniform environmental conditions, (9 h light/15 h darkness). Blood samples were collected seven times over a period of 24 h, every 4 h. Plasma renin activity (PRA), plasma aldosterone (PA), sodium, potassium and chloride concentrations were determined. PRA and PA of both groups changed during 24 h, with the highest values in the dark phase and with higher RAA system activity (especially during the night) in the pregnant goats. In the non-pregnant goats, no circadian changes in PRA and PA were observed. The circadian changes in PRA and PA found in pregnant goats had acrophases at 06:27 h and 01:13 h, respectively. Plasma electrolyte concentrations in both groups of goats also changed during 24 h. These results suggest that circadian changes of potassium concentration in plasma of goats during late pregnancy may be one of the main factors affecting the RAA system. PMID- 12643986 TI - Combined effect of ascorbic acid and selenium supplementation on alcohol-induced oxidative stress in guinea pigs. AB - Oxidative stress is a key step in the pathogenesis of ethanol associated liver injury. Ethanol administration induces an increase in lipid peroxidation either by enhancing the production of oxygen reactive species or by decreasing the level of endogenous antioxidants. In this present study, four groups of male guinea pigs (Cavia porcellus) were maintained for 45 days as follows: Control group (1 mg ascorbic acid (AA)/100 g body wt./day); Ethanol group (1 mg AA/100 g body wt./day+900 mg ethanol/100 g body wt./day); Selenium+AA group (25 mg AA+0.05 mg sodium selenite/100 g body wt./day); Ethanol+Se+AA group (25 mg AA+0.05 mg sodium selenite/100 g body wt.+900 mg ethanol/100 g body wt./day). Malondialehyde (MDA), hydroperoxides (HP) and conjugated dienes (CD) were significantly increased, while the activities of scavenging enzymes superoxide dismutase (SOD) and catalase were reduced in the alcohol administered groups. Co-administration of Se+AA along with alcohol increased the activities of scavenging enzymes and reduced the lipid peroxidation products level in hepatic tissues of guinea pigs. Activities of glutathione peroxidase (GPX) and glutathione reductase (GR) were enhanced in co-administered group. gamma-Glutamyl transpeptidase (GGT), a marker enzyme of alcohol induced toxicity, was also reduced, as was the glutathione content. This study suggests that the combined effect of Se+AA, provides protection against alcohol-induced oxidative stress as evidenced from the decreased levels of lipid peroxidation products and enhanced activities of scavenging enzymes. PMID- 12643987 TI - The effect of lead ions on the energy metabolism of human erythrocytes in vitro. AB - The aim of this work was to evaluate the influence of chronic exposure to lead ions on the parameters of energetic status of human erythrocytes in vitro. Umbilical cord erythrocytes were incubated with lead acetate at final lead ion concentrations ranging from 10 to 200 microg/dl. ATP, ADP, AMP, adenosine, GTP, GDP, GMP, guanosine, IMP, inosine, hypoxanthine, NAD and NADP concentrations in erythrocytes were determined using HPLC. Scanning electron micrographs of erythrocytes were taken. The mean concentrations of ATP, GTP, NAD and NADP, and mean values of adenylate energy charge (AEC) and GEC in cells incubated at the presence of lead ions were significantly lower after 20 h of incubation. Concentrations of purine degradation products (Ado, Guo, Ino) and Hyp were significantly higher. It is suggested that lead ions affect the energy metabolism of erythrocytes. Morphological changes in erythrocytes correspond to the increase of lead ions in the incubation mixture and to the decrease of ATP concentration in erythrocytes. A decrease in NAD and ATP concentration in erythrocytes could be a sensitive indicator of energy process disturbance, useful in monitoring in case of chronic lead exposure. PMID- 12643988 TI - The mosquito genome: perspectives and possibilities. AB - Anopheles gambiae is the mosquito vector responsible for transmitting Plasmodium falciparum, a malaria parasite of humans. With the emergence of genome projects for a variety of prokaryotic and eukaryotic microorganisms, there has been a long standing interest in sequencing the genomes of the malaria parasite and its insect vector. This tour de force effort has now been completed and reported. The alignment of putative orthologs in An. gambiae with those of Drosophila melanogaster highlights several similarities and differences. These findings could have implications in: (1) identifying new targets for insecticide development; (2) strengthening our understanding of the developmental biology of mosquitoes; and (3) possibly controlling pathogen transmission. A brief overview of these interesting findings and the implications for further studies will be discussed here. PMID- 12643989 TI - Aping Jane Goodall: insights into human lymphatic filariasis. AB - The relationship between infection and disease has been a subject of intense debate in human filariasis. Most patients with chronic disease such as lymphedema or elephantiasis are free of demonstrable current infection. Based on cross sectional data and a few assumptions and presumptions, two models of natural history of filariasis have been in vogue during the past two decades. The only (but arduous) way to understand the sequence of events is to follow up subjects with and without patent infections over a period of several years. This article offers critical comments and highlights the insights acquired from such studies. PMID- 12643990 TI - Gymnophalloides seoi: a new human intestinal trematode. AB - A new minute intestinal fluke, recovered from a Korean woman suffering from acute pancreatitis and gastrointestinal difficulties, was described as Gymnophalloides seoi. The coastal village where the patient resided was found to be a highly endemic area for this fluke, and 24 other endemic areas have been found. This fluke infection is now recognized as an emerging parasitic disease. Its second intermediate host is the oyster, and migrating birds are a natural definitive host. This new trematode might exist in other countries. PMID- 12643991 TI - Trypanosomiasis control: surmounting diminishing returns. PMID- 12643993 TI - Can primaquine therapy for vivax malaria be improved? AB - The incidence and range of endemic malaria caused by Plasmodium vivax has expanded during the past 30 years. This parasite forms hypnozoites in the liver, creating a persistent reservoir of infection. Primaquine (PQ), introduced 50 years ago, is the only drug available to eliminate hypnozoites. However, lengthy treatment courses and follow-up periods are not conducive to assessing the effectiveness of this drug in preventing relapses. Resistance to standard therapy could be widespread. Studies are urgently needed to gauge this problem and to determine the safety, tolerability and efficacy of shorter courses and higher doses of PQ. PMID- 12643994 TI - Regulation of antigenic variation in Plasmodium falciparum: censoring freedom of expression? AB - Plasmodium falciparum employs a strategy of clonal antigenic variation to evade the host immune response during the intraerythrocytic stage of its life cycle. The major variant parasite molecule is the P. falciparum erythrocyte membrane protein (PfEMP)1, which is encoded by the var multigene family. The parasite switches between different PfEMP1 molecules through regulation of var transcription. Recent studies have shed considerable light on this process, but much remains unknown. However, striking parallels between transcriptional control of var and genes in other organisms provide direction for future studies. PMID- 12643995 TI - The significance of transferrin receptor variation in Trypanosoma brucei. AB - The transferrin receptor of Trypanosoma brucei is encoded by genes located in different expression sites. The various expression sites encode slightly different transferrin receptors, which differ substantially in their affinity for transferrin of different host species. It was proposed that T. brucei has developed multiple expression sites encoding different transferrin receptors not only to cope with the diversity of mammalian transferrins, but also to ensure sufficient iron uptake in the presence of anti-transferrin receptor antibodies. This article shows that calculations based on K(d) values argue against the first part of the hypothesis, but might support the second part. PMID- 12643996 TI - Reversing Wolbachia-based population replacement. AB - Genetic manipulation that reduces the competence of a vector population to transmit pathogens would provide a useful tool to complement current control strategies, which are based primarily on the reduction/exclusion of vector populations and the prophylactic/therapeutic treatment of the vertebrate host population. Genetic drive is an important component of vector population replacement strategies, facilitating the replacement of natural populations with a genetically modified population. Genetic drive is reviewed here, emphasizing strategies that would employ infections of intracellular Wolbachia bacteria as a vehicle for population replacement. Also discussed are strategies for the retarding, arresting or reversing of Wolbachia-based population replacement. These strategies are based upon altering the conditions required for transgene invasion and are a prudent safeguard, should unexpected detrimental effects become associated with transgene spread. PMID- 12643997 TI - The Plasmodium sporozoite journey: a rite of passage. AB - Sporozoites are the most versatile of the invasive stages of the Plasmodium life cycle. During their passage within the mosquito vector and the vertebrate host, sporozoites display diverse behaviors, including gliding locomotion and invasion of, migration through and egress from target cells. At the end of the journey, sporozoites invade hepatocytes and transform into exoerythrocytic stages, marking the transition from the pre-erythrocytic to the erythrocytic part of the life cycle. This article discusses recent work, mostly done with rodent malaria parasites, that has contributed to a better understanding of the sporozoites' complex biology and which has opened up new avenues for future sporozoite research. PMID- 12643998 TI - Human migration, mosquitoes and the evolution of Plasmodium falciparum. AB - To date, coalescent analysis of the Plasmodium falciparum genome sequence has failed to provide a unifying theory regarding the parasite's evolution. While a better understanding of the evolution of the malaria genome will undoubtedly clarify the current controversy, the importance of the parasite's interplay with both the human host and mosquito vector cannot be underestimated. Changes in the population biology or ecology of either one of these species have consequences for malaria transmission and this was never more apparent than in the environmental changes brought about by the advent of agriculture. PMID- 12643999 TI - Estrogen administration in patients with chronic heart failure: not ready for prime time. PMID- 12644000 TI - It cannot be cardiac failure because the heart is not enlarged on the chest X ray. PMID- 12644001 TI - Hypoxia, angiotensin-II, and norepinephrine mediated apoptosis is stimulus specific in canine failed cardiomyocytes: a role for p38 MAPK, Fas-L and cyclin D1. AB - BACKGROUND: Apoptosis may contribute to the myocardial dysfunction associated with heart failure (HF). Activation of the p38 MAPK cascade can induce apoptosis in non-cardiac cells through increased expression of Fas-L, or through decreased expression of cyclin D(1). AIMS: We tested the hypothesis that hypoxia (HX), angiotensin-II (A-II) and norepinephrine (NEPI) can mediate apoptosis by activating p38 MAPK, and thus initiating stimulus specific changes in Fas-L and cyclin D(1) expression in failing cardiomyocytes. METHODS AND RESULTS: Cardiomyocytes isolated from ten dogs with HF induced by coronary microembolizations were subjected to HX or A-II or NEPI with and without a p38 MAPK inhibitor (SB 203580). TUNEL staining for DNA fragmentation and Western blots for p38 MAPK, Fas-L and cyclin D(1) detection were performed. HX-induced apoptosis was associated with increased Fas-L expression, A-II-induced apoptosis was associated with increased Fas-L and decreased cyclin D(1) expression, and NEPI-induced apoptosis was associated with decreased cyclin D(1) expression. Inhibition of p38 MAPK activity attenuated stress-induced apoptosis in all experiments and reversed changes in Fas-L and cyclin D(1) expression. CONCLUSIONS: HX, A-II and NEPI mediate apoptosis in failing cardiomyocytes via different effects on Fas-L and cyclin D(1) expression. Inhibition of p38 MAPK reversed these effects, suggesting that apoptosis induced by HX, A-II and NEPI involves activation of p38 MAPK upstream from Fas-L and cyclin D(1). PMID- 12644002 TI - Altered calcium transient and development of hypertrophy in beta2-adrenoceptor overexpressing mice with and without pressure overload. AB - Transgenic (TG) mice with cardiac specific 200-fold overexpression of beta(2) adrenoceptors (beta(2)-AR) have a facilitated development of heart failure following thoracic aortic constriction (TAC). We have studied the alterations of intracellular Ca(2+) transients and myocyte size in wild-type (WT) and TG mice after TAC. Cardiomyocytes were isolated from mice 9 weeks after TAC or sham operation, and incubated with Fura 2/AM. The Ca(2+) transients were determined by Spex dual wavelength Spectrometer during electrical stimulation. The cell size was also determined planimetrically. Cells of sham operated TG mice displayed higher systolic Ca(2+) amplitude than respective WT group (DeltaF(340)/F(380) ratio: 1.05+/-0.08 vs. 0.63+/-0.05; P<0.01), a finding in keeping with enhanced ventricular contractility in the TG mice. However, hypertrophied and failing myocytes of TG animals showed a fall in Ca(2+) transients from sham-operated control levels and there was no difference between TG and WT groups following TAC. In sham-operated groups, the cell size of TG mice was significantly bigger than in WT animals (3212+/-139 vs. 2605+/-162 microm(2); P<0.05). The cell size increased to a similar extent in both groups after TAC (4715+/-216 vs. 5027+/-365 microm(2), P=n.s.). In summary, hypertrophy of cardiomyocytes was present in beta(2)-AR TG mice under baseline conditions. A further hypertrophy occurred during pressure overload to an extent similar to that in WT animals. However, the increased intracellular Ca(2+) transient, seen in sham-operated TG mice, was no longer detectable following development of severe hypertrophy and heart failure. These findings provide explanation on the lack of hemodynamic benefit in beta(2) AR TG mice subjected to pressure overload. PMID- 12644003 TI - Leukemia inhibitory factor is augmented in the heart in experimental heart failure. AB - Leukemia inhibitory factor (LIF) is a pleiotropic cytokine that induces cardiac myocyte hypertrophy through the signal transducing molecule, glycoprotein 130. To date, localization of LIF in the heart and regulation of cardiac LIF expression in congestive heart failure (CHF) remain undefined. The present study investigates the potential activation of LIF expression in the failing canine heart that was produced by progressive rapid ventricular pacing. Immunohistochemistry for LIF revealed that LIF immunoreactivity was present in the atrial and ventricular myocytes of the normal heart and was markedly increased in the failing heart as compared to the normal heart. Northern blot analysis demonstrated that cardiac LIF mRNA was increased in both atrium and ventricle in CHF as compared to the normal heart (P<0.01). Linear regression analysis revealed a positive correlation between atrial LIF mRNA and atrial pressure (r=0.87, P<0.001 in right atrium and r=0.86, P<0.001 in left atrium). Positive correlations between left ventricular LIF mRNA and left ventricular dimensions (r=0.91, P<0.0001 in end-systolic diameter; r=0.86, P<0.001 in end diastolic diameter), and an inverse correlation between left ventricular LIF mRNA and left ventricular ejection fraction (EF) were observed (r=-0.93, P<0.0001). There was a positive correlation between left ventricular LIF mRNA and left ventricular mass index (LVMI) (r=0.85, P<0.001). The present study demonstrates that cardiac LIF immunoreactivity and its gene expression are increased in a canine model of experimental CHF and suggests a potential role for LIF in the pathophysiology of CHF. PMID- 12644004 TI - Endothelial dysfunction and infarct-size relate to impaired EDHF response in rat experimental chronic heart failure. AB - BACKGROUND: The rat coronary ligation model of chronic heart failure has been extensively used to investigate its pathophysiology including the role of endothelial dysfunction. Inconsistent results have been obtained concerning the role of endothelial dilative mediators nitric oxide (NO) and endothelium-derived hyperpolarizing factor (EDHF). AIMS: Our aim was to investigate involvement of NO and EDHF in aortic endothelial dysfunction in this model and the influence of individual infarct sizes. Furthermore, we investigated whether it is justified to regard rats that failed to develop large infarct sizes as SHAM controls. METHODS: We performed coronary ligations and SHAM operations and studied acetylcholine (ACh)-induced relaxations and underlying endothelial mediators in isolated aortic rings 12 weeks after infarction. By then, cardiac and hemodynamic parameters were deteriorated in animals with large myocardial infarctions (large-MI, 35+/-3%), but not those with small myocardial infarctions (small-MI, 5+/-2%). RESULTS: Large-MI showed decreased ACh-induced relaxation compared to SHAM due to decreased contribution of EDHF which was inversely correlated with individual infarct-size. Interestingly, small-MI showed significantly increased ACh-induced relaxation compared to SHAM due to increased NO contribution. CONCLUSIONS: Our results suggest that impaired aortic endothelial dilatory function in large-MI is mainly due to an impaired EDHF response and strongly depends on individual infarct-size. In addition, endothelium-dependent relaxation of small-MI rats differed from SHAM, indicating that both groups may not be pooled to serve as controls. These results emphasize the importance of infarct-size and choice of the control group, and may explain inconsistencies in previous studies. PMID- 12644006 TI - Tumor necrosis factor-alpha polymorphism in Turkish patients with dilated cardiomyopathy. PMID- 12644005 TI - Bedside B-type natriuretic peptide and functional capacity in chronic heart failure. AB - OBJECTIVES: To determine if B-type natriuretic peptide (BNP) measurement could be useful in determination of functional capacity in patients suffering from chronic heart failure. BACKGROUND: Evaluating functional capacity is a crucial factor in the follow-up of patients with chronic heart failure. There are numerous methods for measuring functional capacity and their relative merits remain under discussion. Clinical classifications are very subjective and other methods are difficult to use in clinical practice. METHODS: We evaluated functional capacity in 151 consecutive patients using the 6-min walk test. All patients were clinically classified using the New York Heart Association (NYHA) classification. We measured BNP plasma levels using a bedside BNP test. RESULTS: Six minute walk test performance decreased through NYHA classes 1 to 4 (469+/-87, 411+/-82, 325+/ 83 and 196+/-63 m, respectively, P<0.01) and BNP levels increased through NYHA classes 1 to 4 (26.3+/-7.2, 73+/-13, 401+/-74 and 924+/-84 pg/ml, respectively, P<0.001). There was a significant correlation between 6-min walk test performance and BNP plasma levels (R=0.69 P<0.001) and a weaker correlation between BNP and left ventricular ejection fraction (R=0.45 P<0.04). In some patients there was a mismatch between NYHA classification and 6-min walk test performance. In all cases BNP could correct the clinical estimation of functional capacity. When we divided the patients into three sub-groups within each NYHA class, we showed that using BNP could better define functional capacity in patients suffering from chronic heart failure in NYHA classes I to III. CONCLUSION: The measurement of BNP levels thus usefully supplements the clinical examination. The existence of bedside BNP testing methods facilitates its use in routine clinical practice. It also permits easier follow-up of patients with chronic heart failure. PMID- 12644007 TI - Distinct left bundle branch block pattern in ischemic and non-ischemic dilated cardiomyopathy. AB - BACKGROUND: A high percentage of patients with dilated cardiomyopathy have the electrocardiographic (ECG) pattern of advanced left bundle branch block (LBBB). In the present study we sought to investigate whether patients with dilated cardiomyopathy of ischemic or non-ischemic etiology can be differentiated on the basis of LBBB pattern. METHODS AND RESULTS: The study population included 41 patients with dilated cardiomyopathy of non-ischemic (NIC) (n=26) or ischemic origin (IC) (n=15) and LBBB on surface ECG. ECG duration and voltage were digitally measured. The presence of notching of S wave in right precordial leads (V1-V3) was not statistically different between the groups. The voltages of precordial leads V2, V3 and the Sigma(V1+V2+V3 voltages) were significantly more prominent in patients with NIC (P=0.002, P<0.001 and P=0.002, respectively). The discriminative power of receiver operating characteristic analysis was best at voltages of V3 of 2100 microV (area under the curve, 0.805; standard error, 0.001). The sensitivity and specificity of V3 voltage >2100 microV on surface ECG in the presence of LBBB to identify a cardiomyopathy of non-ischemic origin were 85 and 73%, respectively. CONCLUSIONS: A single ECG criteria, voltage of lead V3, appears to be a useful parameter to identify patients with dilated cardiomyopathy of ischemic or non-ischemic origin in the presence of advanced LBBB. PMID- 12644008 TI - Anti-oxidative properties of beta-blockers and angiotensin-converting enzyme inhibitors in congestive heart failure. AB - BACKGROUND: Chronic elevation of plasma catecholamines and sympathetic stimulation in chronic heart failure (CHF) leads to increased production of free radicals, and so possibly to endothelial damage/dysfunction and atheroma formation. Abnormal oxidative stress may therefore be related to some of the high mortality and morbidity in CHF. The objective of the present prospective open study was to compare the effects of beta-blockers and ACE inhibitors in relation to oxidative stress and endothelial damage in CHF. METHODS: We studied 66 outpatients with CHF: 46 patients were established on an ACE inhibitor and were then started on a beta-blocker, and 20 patients not previously on ACE-inhibitors were started on lisinopril. Baseline levels of the measured parameters were compared to 22 healthy control subjects. Serum lipid hydroperoxides (LHP) and total antioxidant capacity (TAC) were determined as indices of oxidative damage and antioxidant defence, and plasma von Willebrand factor (vWf) as an index of endothelial damage/dysfunction. RESULTS: Baseline indices for the measures of oxidative damage and endothelial function in the 66 CHF patients were significantly higher than healthy control subjects [median LHP 7.5 (5.9-12.6) vs. 4.8 micromol/l, P=0.0022; TAC 428 (365-567) vs. 336 Trollox Eq. Units, P=0.0005; mean vWf 134+/-27 vs. 89+/-23 IU/dl, P<0.0001]. Following 3 months of maintenance therapy with beta-blockers, there was significant reduction in LHP levels, but not TAC or vWf. ACE inhibitor therapy also significantly reduced vWf levels, but failed to have any statistically significant effects on LHP or TAC. CONCLUSION: This pilot study suggests that oxidative stress in CHF may be due to increased free radical production or inefficient free radical clearance by scavengers. beta Blockers, but not ACE inhibitors, reduced lipid peroxidation in patients with CHF. No relation was demonstrated between a reduction in oxidative damage and endothelial damage/dysfunction. PMID- 12644009 TI - Effect of biventricular pacing on heart rate variability in patients with chronic heart failure. AB - BACKGROUND: Biventricular pacing is emerging as a long-term therapy for symptomatic heart failure. Analysis of heart rate variability (HRV) has become an important predictive tool in this syndrome. AIM OF THE STUDY: To assess whether chronic resynchronization therapy can affect HRV in patients with heart failure. METHODS AND RESULTS: Thirteen patients with heart failure were studied (mean age+/-1 S.E. 65+/-2.2 years, QRS 195+/-5.3 ms, NYHA class 3.2+/-0.1, LVEF 21+/ 1.7%). The protocol included a preliminary no pacing period for 1 month following device implantation. Twenty-four hour Holter ECG recordings were performed at the end of this period (baseline) and after 3 months of biventricular stimulation (VDD mode). Prior to and following pacing patients underwent NYHA class evaluation, 6-min walk test, Quality of Life Assessment and a cardiopulmonary exercise test. Biventricular pacing improved functional class (P<0.0001) and Quality of life (P<0.0001), increased 6-min walk distance, (P=0.008) and exercise duration (P<0.0001) but had no significant effect on peak exercise VO(2). Resynchronization therapy increased mean 24-h RR (922+/-58 vs. 809+/-41 ms at baseline, P=0.006), SDNN (111+/-11 vs. 83+/-8 ms, P=0.003), SDNN-I (56+/-10 vs. 40+/-5 ms, P=0.02), rMSSD (66+/-14 vs. 41+/-8 ms, P=0.003), Total Power (5724+/ 1875 vs. 2074+/-553 ms(2), P=0.03), Ultra Low Frequency Power (1969+/-789 vs. 653+/-405 ms(2), P=0.03) and Very Low Frequency Power (2407+/-561 vs. 902+/-155 ms(2), P=0.004). CONCLUSION: Biventricular pacing in heart failure improves autonomic function by increasing HRV. This may have important prognostic implications. PMID- 12644010 TI - Regional myocardial perfusion during chronic biventricular pacing and after acute change of the pacing mode in patients with congestive heart failure and bundle branch block treated with an atrioventricular sequential biventricular pacemaker. AB - BACKGROUND: Biventricular (BiV) pacing has been found to improve systolic function and exercise tolerance in patients with severe congestive heart failure and bundle branch block. The mechanisms behind this beneficial effect is still not sufficiently clarified. AIM: To evaluate the regional myocardial perfusion (MP) during BiV pacing and after acute change of the pacing mode to conventional dual chamber (DDD) pacing, and single chamber atrial (AAI) pacing in patients with severe congestive heart failure and prolonged QRS width treated with chronic BiV pacing. METHODS AND RESULTS: Fourteen patients (age 63+/-7 years, 13 male) were evaluated 13+/-7 months after implantation of a triple-chamber biventricular pacemaker. MP was quantified with 13N-labeled ammonia positron emission tomography during BiV pacing, DDD pacing, and AAI pacing. MP was assessed in the anterior, lateral, inferior, and septal regions, and the global mean MP was calculated. Clinical assessment was performed before pacemaker implantation and after at least 3 months of BiV pacing including a 6-min walk test (WT), New York Heart Association (NYHA) class functional score and echocardiography. Global mean MP (BiV: 0.65+/-0.20 vs. DDD: 0.65+/-0.21 vs. AAI: 0.65+/-0.18 mlg(-1)min(-1)) and MP in each of the four regions did not differ between the three pacing modes. The patients improved clinically during BiV pacing; 6 min WT increased (338+/-59 vs. 415+/-73 m, P<0.001), NYHA class score improved (class I/II/III/IV: 0/0/11/3 vs. 1/9/2/0, P<0.001), and left ventricular ejection fraction increased (21+/-5 vs. 29+/-8%, P=0.004). CONCLUSION: No differences in regional MP are detectable after chronic BiV pacing when the pacing mode is changed acutely in patients with severe congestive heart failure and bundle branch block. This finding indicates, that the clinical improvement caused by BiV pacing is not associated with any increase in the MP and thereby oxygen demand. PMID- 12644011 TI - The relevance of heart failure severity for treatment with evidence-based pharmacotherapy in general practice. AB - AIMS: Internationally, research indicates that pharmacotherapy for chronic heart failure (CHF) is sub-optimal. Traditionally, assessment of drug use in heart failure has focused on the use of individual agents irrespective of CHF severity. This study investigates drug use for CHF patients in general practice with respect to the available evidence, incorporating both disease severity and the use of combination drug regimes. METHODS AND RESULTS: A cross-sectional survey of 769 Dutch CHF patients was performed as part of IMPROVEMENT of HF study. For each New York Heart Association severity classification the minimum treatment appropriate for the heart failure severity according to the scientific evidence available at the time of the study (1999) was defined. The proportion of patients treated with each drug increased with increasing severity, with the exception of the beta-blockers. Patients with less severe heart failure were approximately four to eight times more likely to receive evidence-based treatment than those with more severe heart failure. DISCUSSION: To assess pharmacological treatment of heart failure, in relation to the available evidence, it is important to take severity into account. While the number of drugs prescribed increased with increasing severity, the use of evidence-based regimes was lower in patients with more severe heart failure. PMID- 12644012 TI - Patterns of pharmacotherapy in patients hospitalised for congestive heart failure. AB - BACKGROUND: In the 1990s, a number of cardiovascular drugs were evaluated in randomised clinical trials. Treatment guidelines for heart failure were modified to include these evidence-based treatments. AIM: To evaluate the impact of new medical treatments for heart failure between 1990 and 1998. METHODS AND RESULTS: A retrospective cohort study of 2764 patients with a first hospital admission for heart failure between 1990 and 1998. The percentage of patients treated with different cardiovascular drugs after hospitalisation was calculated and compared over time. Use of loop diuretics remained steady approximately 80%, digoxin decreased from 57.6 to 42.7%, angiotensin converting enzyme (ACE) inhibitors showed a slight increase from 49.8 to 54.8%, beta-blockers almost tripled from 11.3 to 28.7%, low dose prophylactic acetylsalicylic acid quadrupled from 9.9 to 39.9%. Kaplan-Meier survival estimates showed highest continuation rates of drug treatment for antithrombotics and diuretics, intermediate for digoxin and ACE inhibitors and low for beta-blockers. More than a quarter of the users discontinued beta-blockers in the first year after hospitalisation. CONCLUSIONS: We observed an increase in the prescribing of several important drug classes, reflecting changes in treatment guidelines during the study period. However, our findings show that not all patients were receiving optimal treatment. More research into the reasons for this is warranted. PMID- 12644013 TI - Factors influencing the length of hospital stay of patients with heart failure. AB - INTRODUCTION: Heart failure (HF) is characterised by frequent hospital admissions and prolonged length of hospital stay. Admissions for HF have increased over the last decade while length of stay has decreased; the reasons for this change in length of stay are uncertain. This study investigates the effect of patient related variables, in-hospital progress and complications on length of stay. METHODS: Patients admitted to Auckland Hospital general medical service and randomised into the Auckland Heart Failure Management Programme were included in this study. RESULTS: One hundred and ninety-seven patients were included in this study. Mean age 73 years, mean left ventricular ejection fraction 32%; 52% had one or more previous HF admissions and 75% were New York Heart Association class IV at admission. Median length of hospital stay was 6 days (IQR 4, 9) which is comparable to the national average from New Zealand admission databases. Longer than average length of stay, defined as >6 days, was associated with the presence of peripheral congestion, duration of treatment with intravenous diuretic, the development of renal impairment, other acute medical problems at admission, iatrogenic complications during hospital stay, and social problems requiring intervention. Factors independently associated with length of stay in the top quartile (>10 days) on logistic regression included the presence of oedema at admission (OR 10.5), change in weight during stay (OR 1.3), duration of treatment with iv diuretic (OR 7.5), the development of renal impairment (OR 9.8), concurrent respiratory problems requiring specific treatment (OR 3.8), and social problems requiring intervention (OR 6.8). CONCLUSIONS: Peripheral congestion, concomitant acute medical problems requiring specific treatment, the development of renal impairment and the presence of social problems were related to a longer than average length of hospital stay. Multivariate models only partly explained variance in hospital stay, suggesting the importance of pre-admission and post discharge factors, including the healthcare environment, the availability of primary and secondary care resources, and the threshold for hospital admission. PMID- 12644014 TI - Cytokine modulation of nuclear factor-kappaB activity in B-chronic lymphocytic leukemia. AB - OBJECTIVE: Dysregulation of the apoptotic mechanisms plays a key role in the accumulation of malignant B-chronic lymphocytic leukemia (B-CLL) cells. The transcription nuclear factor (NF)-kappaB is important for cell survival by regulating the expression of anti-apoptotic genes. Several cytokines can modulate leukemic growth and apoptosis in B-CLL. The aim of this study was to determine whether cytokine-mediated regulation of apoptosis occurs via modulation of NF kappaB activity in peripheral blood mononuclear cells from B-CLL patients. PATIENTS AND METHODS: We evaluated NF-kappaB activity in peripheral blood mononuclear cells from 15 untreated B-CLL patients and 11 controls in resting conditions and in the presence of phorbol-12-myristate-13-acetate (PMA) and different cytokines by electrophoretic mobility shift assay. Apoptosis was studied by spectrophotometric analysis of DNA fragmentation. RESULTS: We found a constitutive high NF-kappaB activity not induced by PMA in B-CLL patients, in contrast with a normal inducible NF-kappaB activity in controls. In B-CLL cultures, addition of interleukin (IL)-4 and IL-13 increased, whereas transforming growth factor (TGF)-beta reduced NF-kappaB activity compared with unstimulated cultures. Accordingly, IL-4 and IL-13 decreased, whereas TGF-beta increased DNA fragmentation compared with unstimulated cultures. IL-13 and IL-4 production was increased, whereas TGF-beta was reduced in PMA-stimulated and unstimulated cultures from B-CLL patients compared with controls. CONCLUSIONS: B CLL patients have a constitutive high NF-kappaB activity, which is modulated by cytokines. In particular, TGF-beta displays a pro-apoptotic activity, whereas IL 4 and IL-13 have opposite effects. These cytokine alterations could be responsible for a positive autocrine circuit that maintains leukemic cells in a pre-apoptotic state. PMID- 12644015 TI - Radioprotective effect of FLT3 ligand expression regulated by Egr-1 regulated element on radiation injury of SCID mice. AB - OBJECTIVE: Hematopoietic factors have an important effect on the regulation of hematopoiesis by stimulating the proliferation of hematopoietic progenitor cells. Although the cytokines that stimulate hematopoiesis have also often proved to exert radioprotective effects, no definitive correlation has been found between the expression of these cytokines regulated by radio-inducible genes and their radioprotective effects. In the current experiments, we evaluated the radioprotective effects of the hematopoietic growth factors regulated by a radio inducible promoter on radiation injury. METHODS: The human Flt3 (FL) cDNA and enhanced green fluorescent protein (EGFP) cDNA were linked together with the internal ribosome entry site (IRES) and then inserted into the eukaryotic expression vector pCI-neo with the Egr-1 promoter (Egr-GF), and the vector was transduced into bone marrow stromal cell lines HFCL (HFCL/EGF). The level of green fluorescence in HFCL/EGF cells was detected after radiation with flow cytometry. The expression of FL in irradiated HFCL/EGF cells was confirmed with Northern blot and ELISA. The HFCL/EGF and CD34(+) cells from human umbilical cord blood were sequentially transplanted intravenously into sublethally irradiated severe combined immunodeficient (SCID) mice. The numbers of peripheral white blood cells transplanted into recipient mice were detected. RESULTS: The activity of EGFP in transfected cells was significantly increased after exposure to gamma radiation at 2.0, 2.5, and 5.0 Gy as compared with nontransfected cells. The expression of FL in HFCL/EGF was significantly higher than that of the control groups (HFCL, HFCL/pCI-neo, and HFCL/FG). The level of secreted FL in serum-free supernatants of HFCL/EGF on human CD34(+) cells was higher than that of control groups. In contrast with three control groups (HFCL, HFCL/pCI-neo, and HFCL/GF), HFCL/EGF resulted in a proportional increase in the number of white blood cells at an early stage after radiation. CONCLUSIONS: We show that radiation enhances the ability of expression of FL in HFCL/EGF to stimulate the proliferation of hematopoietic progenitor cells. These results suggest in vivo use of gene therapy of FL regulated by the Egr-1 promoter protects hematopoiesis from irradiation induced damage. PMID- 12644016 TI - Hydroxamide derivatives of short-chain fatty acids are potent inducers of human fetal globin gene expression. AB - OBJECTIVE: To examine whether hydroxamic acids are inducers of fetal hemoglobin expression, we tested the effects on gamma gene expression of butyric and propionic hydroxamic acids and of two other hydroxamic acids (SBHA and SAHA), which are potent inhibitors of histone deacetylase (HDAC). We also investigated whether there is a correlation between HDAC inhibitory activity of the compounds and their ability to induce gamma-globin gene expression. MATERIALS AND METHODS: Effects on gamma-globin expression were assessed by two methods: 1) a screening assay in which specific gamma-globin gene inducers are recognized by their ability to increase gamma firefly luciferase activity significantly more than beta-renilla luciferase activity; and 2) measurements of gamma-globin mRNA and the frequency of fetal hemoglobin-positive erythroblasts in cultures of burst forming unit erythroid (BFU-E) from normal individuals. HDAC in vitro activity was measured with a partially purified rat liver HDAC and a fluorogenic substrate. RESULTS: All compounds tested increased gamma firefly luciferase activity, gamma/gamma+beta mRNA ratios, and percentage of fetal hemoglobin containing erythroblasts in BFU-E cultures, in a dose-dependent fashion. Butyryl hydroxamic acid 100 microM increased the gamma/gamma+beta mRNA ratios by 5.8-fold and the frequency of fetal hemoglobin-containing erythroblasts by 4.1-fold. Propionyl-hydroxamic acid 150 microM increased the gamma/gamma+beta ratios by 6.3 fold and the fetal hemoglobin-containing erythroblasts by 3.9-fold. SBHA induced gamma-globin gene expression at very low concentrations, 5 to 20 microM in the luciferase system and 2 to 8 microM in BFU-E cultures; SAHA at 1 to 7.5 microM in the luciferase system and 1 to 2.5 microM in the BFU-E cultures. HDAC in vitro inhibition was observed in the millimolar range for propionate and butyrate. IC(50) determinations led to values of 384 microM for propionyl-hydroxamate, 47 microM for butyryl-hydroxamate, 0.93 microM for SBHA, and 0.26 microM for SAHA. CONCLUCION: Our data indicate that hydroxamic acid-based HDAC inhibitors are potent gamma-globin gene inducers and that the concentration range of their effects on gamma gene expression can be correlated roughly with their HDAC inhibitory potencies. PMID- 12644017 TI - Genomic p16 abnormalities in the progression of chronic myeloid leukemia into blast crisis: a sequential study in 42 patients. AB - OBJECTIVE: The molecular abnormalities involved in the progression of chronic myeloid leukemia (CML) are poorly understood. Genetic alterations of the INK4A/ARF locus have been implicated in the lymphoid blast crisis (BC), but sequential studies are not available. The aim of this study was to contribute to a better knowledge of the status of such locus in the different phases of CML and to analyze the prognostic significance of its inactivation. MATERIALS AND METHODS: Sequential assessment by quantitative real-time polymerase chain reaction (PCR) and conventional semiquantitative PCR of p16 exon 2 deletions was performed in 42 CML patients in whom paired DNA samples from the chronic phase and the BC were available. Samples of 10 healthy donors and 30 patients with nonleukemic myeloproliferative syndromes served as controls. The methylation status of the promoter region of the p16 gene was also studied by methylation specific PCR. RESULTS: The concordance rate between the two PCR techniques was 97.8% (87/89). By real-time PCR, homozygous p16 deletions were found in 6 of 21 patients (29%) with lymphoid BC, whereas they were not observed in chronic-phase CML nor in 21 myeloid BC patients. Hypermethylation of the p16 gene was not detected in any of the lymphoid BC. No specific clinical profile was associated with homozygous p16 deletions. Therapeutic response and survival did not significantly differ in p16-deleted and p16 germline lymphoid BC patients. CONCLUSION: P16 gene deletions are detected in a substantial proportion of lymphoid BC of CML by quantitative real-time PCR analysis, but this is not associated with any clinico-hematological feature other than lymphoid phenotype and does not influence the patients' outcome. Such technique is simple and reliable to assess the p16 gene status. PMID- 12644018 TI - The role of mitochondrial-mediated apoptosis in a myelodysplastic syndrome secondary to congenital deletion of the short arm of chromosome 4. AB - OBJECTIVE: Myelodysplastic syndromes (MDS) are characterized by peripheral cytopenia and ineffective hematopoiesis. In adult-onset MDS and in certain inherited marrow failure syndromes, apoptosis is increased and is mediated mainly through activation of the Fas pathway. It is unclear whether the various myelodysplastic disorders share the same apoptosis pathways. I investigated apoptosis pathways in a patient with refractory cytopenia with ring sideroblasts associated with congenital 4p deletion to determine the mechanism for bone marrow failure. METHODS: Marrow cells and lymphoblast cell lines generated from peripheral blood were analyzed for apoptosis and protein expression by flow cytometry, Western blot, and confocal microscopy, either directly or after gamma irradiation (15 G). Cell viability after treatment with inhibitors of specific apoptosis pathways was also determined. RESULTS: Compared to controls, the patient's marrow and lymphoblastoid cells showed significantly higher apoptosis rates and activation of caspase-3. Investigation of the mitochondrial apoptosis pathway showed a consistent pro-apoptosis profile, namely, upregulation of Bax, Bax-alpha, cytochrome c, and Apaf1, and low bcl-2. Differences between the patient's and the normal cells were further accentuated after irradiation; p53 expression was strikingly higher in the patient only after irradiation. In contrast, Fas and FADD expression on the patient's and the control's cells were comparable. Addition of caspase 3 or caspase 9 inhibitors markedly increased patient cell viablity, but blocking anti-Fas antibody did not. CONCLUSION: The ineffective hematopoiesis in this case is explained by increased apoptosis and is linked to hyperactivation of the mitochondrial cell death machinery and not to the Fas pathway, which might be secondary to an intramitochondrial defect. This information is crucial because the development of anti-apoptotic agents for the treatment of MDS may not be universally efficacious and should target the specific derangement. PMID- 12644019 TI - Cycling B-CLL cells are highly susceptible to inhibition of the proteasome: involvement of p27, early D-type cyclins, Bax, and caspase-dependent and independent pathways. AB - OBJECTIVE: Although peripheral blood B-CLL cells are arrested in G0 phase of the cell cycle, a proliferating pool of cells in proliferation centers might be involved in disease progression. We have previously described an in vitro model of this proliferating pool of cells using B-CLL cells stimulated with immunostimulatory oligonucleotides (CpG-ODN) and interleukin-2. Lactacystin is a specific inhibitor of the proteasome and is a potent apoptosis inductor in resting peripheral B-CLL cells. In the present study, we investigated the effect of proteasome inhibition in proliferating B-CLL cells. METHODS: The effect of proteasome inhibition was analyzed using thymidine incorporation, annexin V assays, and TUNEL staining. Immunoblots were performed to evaluate expression of proteins involved in cell cycle and apoptosis regulation. RESULTS: Lactacystin blocked cell cycle progression in activated B-CLL cells and inhibited degradation of p27. Upregulation of cyclin D2 and D3 in activated B-CLL cells was inhibited while the expression of cdk2, cdk4, and cyclin E remained unchanged. Activated B CLL cells were more susceptible to apoptosis induction as compared to resting B CLL cells. Apoptosis induction was accompanied by cleavage of Bax, procaspase 8, procaspase 9, and procaspase 3. However, a broad-spectrum caspase inhibitor (z VAD.fmk) only partially inhibited cell death although DNA degradation was completely inhibited. CONCLUSION: Proteasome inhibition is highly effective in proliferating B-CLL cells and induces apoptosis using a caspase-dependent and independent pathway. PMID- 12644020 TI - Hammerhead ribozyme-mediated cleavage of the fusion transcript NPM-ALK associated with anaplastic large-cell lymphoma. AB - OBJECTIVE: Approximately 60% of all anaplastic large-cell lymphomas (ALCL) contain a specific t(2;5)(p23;q35) chromosomal translocation leading to overexpression of NPM-ALK. As the chimeric tyrosine kinase is involved in tumorigenesis and pathogenesis of ALCL, we were interested to inhibit NPM-ALK expression using an exogenous and an endogenous ribozyme approach. METHODS: We designed five anti-ALK hammerhead ribozymes that were targeted to cleave the ALK proportion of NPM-ALK. The ribozyme with the highest cleavage activity was used as a modified RNA/DNA chimera (RZ1*) for transient transfection and as a self splicing ribozyme vector (pRZ1) for endogenous expression. Ribozyme performance was tested in 293 cells (cotransfected with NPM-ALK) and in the ALCL cell line Karpas 299 by transient and stable transfection and Western blotting. The half life time of NPM-ALK was determined by pulse-chase experiments. RESULTS: In vitro cleavage assays demonstrated different catalytic efficiencies depending on the targeted site of the substrate. Constant transfection of Karpas 299 cells with RZ1* for 96 hours did not lead to a significant reduction of NPM-ALK protein, presumably due to the long half-life of NPM-ALK (48 hours). In contrast, NPM-ALK protein expression was almost completely suppressed in transiently transfected 293 cells. Stable transfection of Karpas 299 cells with pRZ1 also resulted in significant reduction of NPM-ALK expression. CONCLUSION: These results suggest that ribozymes targeted against NPM-ALK are able to inhibit expression of this oncogenic kinase efficiently and will be a useful tool to analyze its role in the pathophysiology of ALCL. PMID- 12644021 TI - Mechanisms of anemia in SHP-1 protein tyrosine phosphatase-deficient "viable motheaten" mice. AB - OBJECTIVE: Viable motheaten mice (abbreviated gene symbol me(v)) are deficient in SHP-1, a critical negative regulator of signal transduction in hematopoietic cells. These mice exhibit severe immune dysfunction accompanied by hyperproliferation of myeloid cells, widespread inflammatory lesions, and regenerative anemia. The aim of this study was to investigate the mechanisms underlying anemia in me(v)/me(v) mice. MATERIALS AND METHODS: Multiple hematologic parameters, osmotic fragility, and erythropoietin levels were measured to characterize the anemia in me(v)/me(v) mice. B-cell-deficient me(v)/me(v) Igh-6(null) mice were generated to assess the role of anti erythrocyte antibodies. Coombs assays and flow cytometry were carried out for detection of anti-erythrocyte antibodies. Oxidant production by macrophages, glutathione levels, and lipid peroxidation products in erythrocytes were measured, as was the impact of oxidant on the ultrastructure of me(v)/me(v) erythrocytes. Erythroid maturation and erythrocyte plasma membrane integrity were assessed with flow cytometry by evaluating CD71 expression and annexin V labeling. RESULTS: The regenerative anemia of me(v)/me(v) mice was associated with erythrocyte changes that were independent of the presence of anti erythrocyte antibodies. Erythrocytes from me(v)/me(v) mice had increased fragility and heightened susceptibility to oxidant damage. Macrophages from me(v)/me(v) mice demonstrated a higher basal level of oxidant production and enhanced production after stimulation. Oxidant damage in me(v)/me(v) erythrocytes was evidenced by a significant elevation of lipid peroxidation and diminished levels of glutathione. CONCLUSION: Our results support the hypothesis that as a consequence of severe inflammatory disease, me(v)/me(v) erythrocytes are subject to exceptionally high oxidative stress resulting in oxidation of phospholipids in the erythrocyte membrane with subsequent hemolysis. PMID- 12644022 TI - Sca+CD34- murine side population cells are highly enriched for primitive stem cells. AB - OBJECTIVE: The aim of this study was to characterize murine side population (SP) stem cells and SP cell subpopulations for primitive stem cell capacity. MATERIALS AND METHODS: SP cells, characterized by a specific Hoechst dye efflux pattern, were isolated by flow cytometric analysis and sorting from murine adult whole bone marrow (WBM). Different subpopulations of SP cells were isolated by staining with anti-Sca and anti-CD34 antibodies. Primitive stem cell content of SP cells and SP subsets were determined by cobblestone area-forming cell (CAFC) frequencies. RESULTS: Measurement of CAFC frequencies revealed that SP cells are greatly enriched for both primitive stem cells (day-28-35 CAFC) and somewhat more mature hematopoietic cells (day-14-21 CAFC) compared to WBM. The day-28 and day 35 CAFC enrichments in SP cells vs WBM cells were 1065 and 471, respectively. Analysis of the subpopulations of SP cells revealed that SP(+)Sca(-)CD34(+) cells contained almost exclusively day-7 CAFC and had little day-28-35 CAFC activity. SP(+)Sca(+)CD34(+) cells had high day-7-14 CAFC frequencies, but lower day-35 CAFC frequencies compared to SP(+)Sca(+)CD34(-) cells. SP(+)Sca(+)CD34(-) cells contained very low day-7 CAFC activity, but nearly 2200 times the day-28-35 CAFC activity as normal bone marrow. To evaluate the influence of Hoechst dye efflux capacity, we divided the SP tail into four groups of cells. The SP cells with lowest efflux of Hoechst dye contained the highest progenitor activity (day-7-14 CAFC). The highest day-35 CAFC frequencies, nearly 6000 times those of normal marrow, were seen in the SP cells with the greatest efflux of the Hoechst dye. CONCLUSIONS: Murine SP cells contain both progenitor and primitive populations of hematopoietic stem cells. The most primitive stem cells measured in the in vitro CAFC assay mark for Sca(+) and CD34(-) and have a high ability to efflux Hoechst dye. Isolation of these cells may provide the means to directly study mechanisms of primitive stem cell damage. PMID- 12644023 TI - Profiling of differentially expressed apoptosis-related genes by cDNA arrays in human cord blood CD34+ cells treated with etoposide. AB - OBJECTIVE: Understanding the molecular events that contribute to survival of and drug-induced apoptosis in hematopoietic stem and progenitor cells (HSC/P) can have impact on more rational approaches to blood cancer therapeutic design, as well as on strategies to minimize toxic side effects of chemotherapeutic drugs. Here we sought to systematically evaluate the basic molecular components and main pathways that govern and mediate cellular response initiated within human CD34(+) cells to etoposide-induced apoptosis. MATERIALS AND METHODS: Human CD34(+) cells were isolated from umbilical cord blood (CB) and expanded in vitro. Expression of apoptosis-related genes in the control and etoposide treated cells was determined using cDNA array and quantitative real-time RT-PCR. RESULTS: We identified a set of apoptosis-related genes expressed in highly purified normal human CB CD34(+) cells and determined how the expression of these genes changed in response to etoposide treatment. In addition, TRAIL does not induce apoptosis of normal human CD34(+) cells, and it has no cytotoxic effect on human CD34(+) cells that are undergoing apoptosis in response to growth factor withdrawal. This may be due to upregulation of cytotoxic receptors as well as the decoy receptor for TRAIL, and c-FLIP. CONCLUSION: p53, c-Myc, and BAFF pathways are main pathways utilized by CD34(+) cells to arrest cell-cycle progression at multiple checkpoints, to halt proliferation, and to induce apoptosis as part of their cellular response to etoposide. Multiple known pro-survival and pro-apoptotic pathways are simultaneously activated in etoposide-treated CD34(+) cells. Also, TRAIL, used alone or in concert with chemotherapeutic drugs, may be of use as a safe blood cancer therapeutic with no or low toxicity for HSC/P. PMID- 12644025 TI - Epidemiology and pathophysiology of symptomatic gastroesophageal reflux disease. AB - Symptomatic, or nonerosive, gastroesophageal reflux disease (GERD) is the most common presentation of GERD. Patients with symptomatic GERD more frequently are younger, female, weigh less, and are less likely to have a hiatal hernia compared with patients who have erosive GERD. Physiologically, these patients demonstrate minimal esophageal motor abnormalities. However, despite a common clinical presentation and similar endoscopic findings, symptomatic GERD is comprised of a heterogeneous group of patients. There are several identifiable subgroups differentiated by the underlying mechanisms causing their heartburn symptoms, distinctions that may explain the relatively low symptom response rate to potent antireflux treatment observed among these patients as compared with those with erosive esophagitis. PMID- 12644024 TI - Ex vivo expansion of human adult stem cells capable of primary and secondary hemopoietic reconstitution. AB - OBJECTIVE: Ex vivo expansion of human hemopoietic stem cells (HSC) is an important issue in transplantation and gene therapy. Encouraging results have been obtained with cord blood, where extensive amplification of primitive progenitors was observed. So far, this goal has been elusive with adult cells, in which amplification of committed and mature cells, but not of long-term repopulating cells, has been described. METHODS: Adult normal bone marrow (BM) and mobilized peripheral blood (MPB) CD34(+) cells were cultured in a stroma-free liquid culture in the presence of Flt-3 ligand (FL), thrombopoietin (TPO), stem cell factor (SCF), interleukin-6 (IL-6), or interleukin-3 (IL-3). Suitable aliquots of cells were used to monitor cell production, clonogenic activity, LTC IC output, and in vivo repopulating capacity. RESULTS: Here we report that BM and MPB HSC can be cultured in the presence of FL, TPO, SCF, and IL-6 for up to 10 weeks, during which time they proliferate and produce large numbers of committed progenitors (up to 3000-fold). Primitive NOD/SCID mouse repopulating stem cells (SRC) are expanded sixfold after 3 weeks (by limiting dilution studies) and retain the ability to repopulate secondary NOD/SCID mice after serial transplants. Substitution of IL-6 with IL-3 leads to a similarly high production of committed and differentiated cells but only to a transient (1 week) expansion of SRC(s), which do not possess secondary repopulation capacity. CONCLUSION: We report evidence to show that under appropriate culture conditions, adult human SRC can also be induced to expand with limited differentiation. PMID- 12644026 TI - The economic and quality-of-life impact of symptomatic gastroesophageal reflux disease. AB - Gastroesophageal reflux disease (GERD) is one of the most common complaints in general medical practice. Symptoms of GERD have a high prevalence, which greatly affects health economics and patient health-related quality of life (HRQOL). The majority of patients who present with reflux symptoms lack esophageal mucosal injury and thus have symptomatic GERD. There appear to be no differences in either symptoms or HRQOL measures between patients with or without positive endoscopy. Although most pharmacoeconomic and HRQOL assessments have focused on erosive GERD patients, available data suggest that the impact of symptomatic GERD on HRQOL is significant and equivalent to that associated with erosive disease. Proton pump inhibitors (PPIs) improve HRQOL to a greater extent than either histamine(2)-receptor antagonists or prokinetics in patients with GERD, with PPI based step-down or "on-demand" strategies yielding the most cost-effective benefit to health. Additional HRQOL and outcomes research focused on patients with symptomatic GERD, ideally using disease-specific assessment tools, is needed. Furthermore, these management strategies must be implemented in real world settings to validate their effectiveness and firmly establish claims of reduced need for diagnostic testing and decreased direct medical costs. PMID- 12644027 TI - Diagnosis of symptomatic gastroesophageal reflux disease. AB - Gastroesophageal reflux disease (GERD) encompasses a heterogeneous group of patients with differences in sensitivity to esophageal acid exposure, perception of pain, and physiological tissue involvement. The most difficult patients to understand are those who have a great symptom burden but no endoscopic evidence of esophageal mucosal involvement. These patients with symptomatic GERD (also called nonerosive reflux disease and endoscopy-negative reflux disease) present a diagnostic challenge. Diagnostic tests, like ambulatory pH monitoring, the acid perfusion test, and intraesophageal balloon distension, have limited reliability in patients with this form of GERD whose symptoms may exhibit poor correlation with acid exposure or mechanostimulation. The recent interest in a proton pump inhibitor therapeutic trial to identify the group of symptomatic GERD patients (having ruled out risk factors for more morbid conditions) who will respond to these agents has considerable appeal. It has been shown effective in about 75% of patients, and offers a simple approach to managing the difficult-to-diagnose patient even while further diagnostic procedures are carried out. PMID- 12644028 TI - Factors that influence therapeutic outcomes in symptomatic gastroesophageal reflux disease. AB - The term "symptomatic gastroesophageal reflux disease" (GERD) refers to those patients who present with the typical GERD symptoms of heartburn and regurgitation, yet do not have endoscopic evidence of esophagitis. The primary goals of managing symptomatic GERD are to control symptoms and improve quality of life. A clinical assessment of the GERD patient can identify important clinical features, such as atypical and extraesophageal symptoms for which acid suppressive agents tend to be less effective. Performing an endoscopy can further identify the patient as having nonerosive reflux disease, erosive esophagitis, or Barrett's esophagus-diagnoses which can help determine treatment but may not prove predictive of therapeutic response. Determining acid exposure through pH testing can predict therapeutic response, with those revealing an abnormal acid exposure time being more responsive to acid-suppressive therapy. However, the performance of an endoscopy and pH testing on each patient is clearly not practical. Whereas the natural history of symptomatic GERD is still largely undefined, acid-suppressive therapy appears to be the best approach available for both the short-term and long-term management of this disease. PMID- 12644029 TI - Goals of therapy and guidelines for treatment success in symptomatic gastroesophageal reflux disease patients. AB - Important new data have established symptomatic gastroesophageal reflux disease (GERD) as a disorder in its own right. Despite major advances in management, this disorder still presents many challenges. With the absence of visible disease (i.e., esophageal erosions), symptom relief must be a primary aim in treating patients with symptomatic GERD; in particular, reflux patients suffer from several different symptoms, but relief of heartburn and acid regurgitation should be the principal concern in clinical practice. Epigastric pain and other upper GI symptoms usually improve with active treatment. From a methodological standpoint, complete symptom relief is an attractive outcome measure. However, not all patients expect complete absence of symptoms in the long term, and many reflux patients are willing to continue a treatment strategy that provides substantial but less than absolute symptom control. Patients with symptomatic GERD have significantly impaired health-related quality of life (HRQOL) which, when heartburn is resolved, often improves to levels equal to or better than that found in the normal, healthy population. However, some reflux sufferers may not report symptoms and have apparently normal HRQOL measures owing only to the fact that they self-impose rigorous lifestyle restrictions to prevent heartburn restrictions that may, in turn, eventually lead to HRQOL impairment. A full and complete evaluation of treatment success should be able to detect this pattern. Data on the long-term prognosis and risk of complications associated with treatment for symptomatic GERD are scarce and incomplete. However, the prevention of erosive esophageal damage, strictures, and adenocarcinoma should be a goal of long-term treatment success, even in symptomatic GERD. PMID- 12644030 TI - Intermittent and on-demand use of proton pump inhibitors in the management of symptomatic gastroesophageal reflux disease. AB - The epidemic of gastroesophageal reflux disease (GERD) in industrialized nations is currently spreading to less-developed ones, with more than half of the patients having symptomatic or mild erosive GERD. The long-term management of GERD has been dominated by daily maintenance treatment with proton pump inhibitors (PPI) to prevent relapse. It is common, however, for many patients with mild disease and infrequent symptom relapses to use a PPI only when symptoms demand. Patients with symptomatic or mild erosive GERD are therefore ideal for on demand or intermittent treatment. The efficacy of such a strategy of intermittent treatment, or treatment of symptoms on demand, has recently been evaluated in four randomized controlled studies. These trials demonstrate that such therapeutic strategies reduce symptoms, improve quality of life, and are cost effective. In clinical practice, the author has found these treatment strategies suitable for approximately 60% of newly diagnosed patients with GERD for the long term management of symptomatic GERD of mild or moderate severity. PMID- 12644032 TI - Diagnosis of drug-induced liver injury in Japanese patients by criteria of Consensus Meetings in Europe. AB - Eighty-five Japanese patients with drug-induced liver injury were assessed by criteria of Consensus Meetings in Europe on causality assessment of drug-induced liver injury. Histopathological investigation was performed for all patients to confirm the diagnosis. We divided these patients into two groups by the date of disease onset. Cases before 1989 were defined as past cases, and those after 1990 as recent cases, because the clinical-pathological characteristics of drug induced liver injury have changed due to the ability to diagnose hepatitis C virus infection since 1990. Fifty-seven patients with drug-induced liver injury were enrolled as past cases, and 28 as recent cases. For past cases, the results of assessment by the criteria of Consensus Meetings in Europe were as follows: 'very likely': 14 patients (25%), 'likely': 23 patients (40%), 'possible': 15 patients (26%), 'dubious': five patients (9%), and 'unlikely': none (0%). For recent cases, the results were as follows: 'very likely': six patients (22%), 'likely': 14 patients (42%), 'possible': six patients (22%), 'dubious': two patients (7%), and 'unlikely': none (0%). There were no differences between the past and recent cases in distribution of assessment. More than 90% of patients were assessed as 'possible' or more, and the remaining seven patients were assessed as 'dubious'. No patients were assessed as 'unlikely'. Five of seven patients assessed as 'dubious' had long-term cholestasis, and two had alcohol consumption. These results indicated that the criteria of Consensus Meetings in Europe were useful for diagnosing drug-induced liver injury in Japanese patients. PMID- 12644033 TI - Effects of alanine in patients with advanced primary biliary cirrhosis: preliminary report. AB - When rats given D-galactosamine are then treated with the glucogenic amino acid alanine, their alanine aminotransferase (ALT) activity, total bilirubin level, and survival rate improve compared with when other amino acids are used. Here, we report a preliminary study of the clinical and pharmacological effects of alanine given to three patients with primary biliary cirrhosis (PBC). The patients were jaundiced and were in the end-stage of the disease. The treatment they had been receiving was continued while they were given 18 g of alanine per day for a planned 8 weeks. For all three patients, test results for total bilirubin, alkaline phosphatase, and ALT decreased by 25% or more from the base line at some time during treatment. The arterial ketone-body ratio increased. Two of the patients reported that their itching and fatigue lessened. Except for one patient given a second course, who reported nausea, adverse effects were not found. In end-stage PBC, alanine administration decreased the total bilirubin level and improved symptoms, so this compound may decrease jaundice in this disease. A long term study of a larger group of patients is needed. PMID- 12644031 TI - Rabeprazole-based therapy in the management of symptomatic gastroesophageal reflux disease. AB - Gastroesophageal reflux disease (GERD) is associated with several symptoms, such as heartburn, belching, and regurgitation, which arise from esophageal exposure to gastric acid. Symptoms may occur in the absence of endoscopically observed esophageal mucosal damage and inflammation. These patients represent the majority of those who present with GERD symptoms. Although acid suppression therapy is a logical approach to relieving GERD symptoms, it has been thought to relieve symptoms less reliably in patients with endoscopically negative, or symptomatic GERD than in those with erosive GERD. Two multicenter, randomized, double-blind, placebo-controlled clinical trials were conducted in the United States to evaluate the efficacy of rabeprazole sodium 10 mg and 20 mg compared with placebo for the relief of heartburn and other symptoms associated with symptomatic GERD. Results from these studies indicated that rabeprazole 10 or 20 mg once daily relieved heartburn within the first 1 or 2 days of treatment and also had significant positive effects on other GERD symptoms, including regurgitation, belching, bloating, satiety, and nausea. Overall, these results suggest that rabeprazole may hold a significant therapeutic advantage in the treatment of heartburn and other symptoms associated with endoscopically negative GERD, particularly in the majority of patients who often are treated empirically without, or before, endoscopic evaluation. PMID- 12644034 TI - Serum levels of gamma-globulin and total bilirubin influence the prevalence of multiple extrahepatic complication in patients with hepatitis C virus infection. AB - Patients with chronic hepatitis C virus (HCV) infection develop various extrahepatic complications. In this study, we analyzed as following. (1) The prevalence and interrelation of multiple extrahepatic manifestations. (2) Exact factors that are related to multiple extrahepatic manifestations. Consecutive autopsies of 261 cases with HCV infection that had undergone autopsy in 1988-1998 were enrolled. One hundred and forty consecutive patients who had gastro enteric carcinoma and underwent autopsy after death in our institution during the same period were used as controls. These 140 patients did not have hepatitis virus and chronic hepatitis or liver cirrhosis histologically. The prevalence of glomerulonephritis (GN), sialadenitis, thyroiditis, and idiopathic pulmonary fibrosis (IPF) was determined histologically. 'Multiple extrahepatic complications' was defined as the presence of two or more of the above four extrahepatic complications. Multiple logistic regression analyses of the risk for the multiple extrahepatic manifestations were performed. GN was present in 58.6% (153 cases), sialadenitis in 16.1% (42 cases), thyroiditis in 7.7% (20 cases), and IPF in 5% (13 cases) in cases of HCV infection. In addition, 37 out of 261 (14.2%) and seven (2.7%) cases had two and three extrahepatic complications, respectively. Multivariate analysis showed that multiple extrahepatic complications correlated with high serum gamma-globulin levels (>/=2.5 g/dl) and high serum total bilirubin levels (>/=1.2 mg/dl) about 1 year before death. Moreover, about 70-80% of patients with sialadenitis, IPF, or thyroiditis had other extrahepatic manifestations. HCV-positive patients with high serum gamma globulin or high total bilirubin are at high risk of multiple extrahepatic complications. PMID- 12644036 TI - Cholestasis as presenting symptom of portal cavernoma. AB - A 51-year-old man with a history of portal vein thrombosis, was examined because of elevated liver tests and a tumoral mass in the liver hilus. Computed tomography (CT) scan and magnetic resonance imaging confirmed the portal vein thrombosis and an infiltrating mass in the porta hepatis with compression on the common bile duct. Endoscopic retrograde cholangiography showed an irregular narrowing of the mid-part of the common bile duct. The patient was referred for explorative laparotomy, which revealed a hypervascular mass in the liver hilus surrounded by many blood vessels. The diagnosis of portal cavernoma was made. Further haematological examination for the cause of portal vein thrombosis revealed an anti-phospholipid syndrome as well as myeloproliferative disease. Oral anticoagulant treatment is started. In conclusion, we report a case of biliary stricture due to portal vein thrombosis and cavernoma (portal biliopathy) which was not diagnosed preoperatively. Biliary strictures associated with portal vein thrombosis are due to extrinsic compression by collaterals and can also be induced by ischemic injury secondary to venous and arterial thrombosis of the choledochal vascular plexus. PMID- 12644035 TI - High-dose interferon-alpha therapy lowers the levels of serum fibrogenesis markers over 5 years in chronic hepatitis C. AB - We examined the levels of serum N-terminal peptide of type III procollagen (P-III NP) and the 7S domain of type IV collagen (IV-7S) as fibrogenesis markers in patients with chronic hepatitis C to clarify whether high-dose interferon-alpha (IFN-alpha) therapy has a suppressive effect on hepatic fibrogenesis for a long period (over 5 years) after the cessation of IFN therapy. Eighty patients with CHC were given 10 million units of IFN-alpha2b daily for 14 days followed by three times per week for a total of 24 weeks. Patients were divided into the following three groups according to the highest serum alanine aminotransferase levels during 1 year observation after the end of IFN therapy: complete responders (CR), partial responders (PR), and nonresponders (NR). We measured serum fibrogenesis markers before and at the end of IFN therapy, and again 1 year and more than 5 years after the end of IFN therapy. Liver biopsies were performed before IFN therapy in all patients and again over long-term observation in 10 patients (PR; 5 and NR; 5). Serum P-III-NP levels significantly decreased after IFN therapy in all three groups of patients, and further decreased in CR and PR over long-term observation. Serum IV-7S levels in CR significantly decreased after IFN therapy and further decreased over long-term observation. Serum IV-7S levels over long-term observation were significantly lower than those at the end of IFN therapy in CR and PR and significantly lower than the initial values in all three groups of patients. The progression of fibrosis was not significant over long-term observation in liver biopsy specimens of 10 patients. The results of the present study suggest that high-dose IFN-alpha therapy for 6 months suppresses the progression of hepatic fibrosis for more than 5 years not only in CR but also in PR. PMID- 12644037 TI - Hydrophilic bile salts have a cytoprotective effect against cyclosporine A induced cholestasis through enhanced canalicular membrane fluidity and transporter activity. AB - Cyclosporine A (CsA) reduces liver canalicular membrane (CM) fluidity to cause a disproportionate reduction of biliary lipid secretion (the uncoupling phenomenon) without affecting adenosine triphosphate-dependent (ABC) transporters except for Mdr1. This study investigated whether hydrophilic bile salts inhibit CsA-induced cholestasis, focusing on CM fluidity and ABC transporter expression. Male Sprague Dawley rats were infused with taurocholate (TC) (200 nmol/min/100 g body weight) for 2 h, flowed by infusion with tauroursodeoxycholate (TUDC), tauroalphamuricholate, or taurobetamuricholate (100 nmol/min/100 g body weight plus TC at 100 nmol/min/100 g body weight). Thereafter, CsA (20 mg/kg body weight) was injected as a bolus and bile was collected for 2 h. Canalicular membrane vesicles were prepared for analysis of cholesterol (CH), phospholipid (PL), CM fluidity, and expression of ABC transporters (Mdr1, Bsep, Mdr2, and Mrp2). CsA administration reduced biliary lipid secretion along with a disproportionately smaller decline of bile salt secretion. Hydrophilic bile salts significantly inhibited cholestasis after CsA injection by increasing CM fluidity and by increasing the expression of Mrp2 and Bsep, whereas Mdr1 and Mdr2 were unaltered. (1) Hydrophilic bile salts inhibit CsA-induced cholestasis, presumably by increasing CM fluidity, and this action was greatest with TUDC. (2) The fact that ABC transporters, except for Mdr1 and Mdr2, were overexpressed in the CM after infusion of these bile salts suggests that cytoprotective bile salts functions may increase transporter mass as well as enhancing transporter activity. PMID- 12644038 TI - Effect of dexamethasone on biliary excretion of bile acids and organic anions in rats. AB - Multidrug resistance protein 2 (Mrp2) is an important transporter for biliary excretion of organic anions, and is reported to be up-regulated by dexamethasone. In the present study, effect of dexamethazone (1 mg/kg) on biliary excretion of bile acids and organic anions was studied in rats. After bile duct cannulation, bile acids and organic anions were intravenously administered, and their biliary excretion was studied. Biliary excretion of tracer doses of taurocholate and taurolithocholate-sulfate was unchanged with dexamethasone. Dexamethasone increased the excretory maximum of taurolithocholate-sulfate and sulfobromophthalein to 1.8 and 1.5 times, respectively, whereas it did not change the excretory maximum of phenolphthalein-glucuronide and taurocholate. Dexamethasone also increased biliary glutathione excretion. These results obtained by in vivo studies can be explained by the up-regulation of Mrp2, which had been reported in in vitro studies. PMID- 12644039 TI - Evaluation of hemodynamics in esophageal varices. Value of endoscopic color Doppler ultrasonography with a galactose-based contrast agent. AB - Endoscopic color Doppler ultrasonography (ECDUS) is a useful modality for obtaining color flow images of esophageal varices. Levovist is a microbubble echo enhancing agent, which improves Doppler ultrasound examination. In this paper, we compared vessel images detected with pre-contrast with those detected by enhanced ECDUS using Levovist in 62 patients with esophageal varices. Color flow images of esophageal varices and paraesophageal veins by pre-contrast ECDUS were obtained in 62 (100%) of the 62 patients. Vessel images of perforating veins were obtained in 48 (77.4%) of the 62 patients with pre-contrast ECDUS. The perforating veins were classified into three types according to the flow direction. Type 1 showed in-flow from the paraesophageal veins to the esophageal varices, Type 2 showed out-flow from the esophageal varices to the paraesophageal veins, and Type 3 was a mixed type showing both in-flow and out-flow. After Levovist contrast, color flow images detected with pre-contrast ECDUS were improved in all patients. Color flow images of perforating veins were obtained in 60 (96.8%) of the 62 patients by enhanced ECDUS using Levovist. Pre-contrast-diagnosed Type 2 patterns had been re-diagnosed as Type 3 in five patients, and three cases with Type 1 had been relabeled as Type 3 pattern using Levovist. The use of Levovist contrast can improve the diagnostic quality of ECDUS examinations of esophageal variceal patients, yielding a much clearer picture of the hemodynamics. PMID- 12644040 TI - Clinicalpathological analysis of risk factors for the development of hepatocellular carcinoma after surgery for esophageal varices due to underlying cirrhosis or pre-cirrhosis in the 397 patients. AB - Risk factors for the development of hepatocellular carcinoma (HCC) were investigated in 397 patients who underwent non-shunt operation for esophageal varices due to underlying cirrhosis or pre-cirrhosis between September 1979 and May 1995. Ninety-five of these patients developed HCC. The clinical characteristics of patients at the time of surgery for varices, stages (F0-F4) of the progression of fibrosis, and grades (A0-A3) of necroinflammatory activity in liver biopsy tissue obtained at surgery in 170 patients based on the New Inuyama Classification (Int. Hepatol. Commun. 6 (1996) 112), were analyzed to investigate their relationship with the development of HCC. In addition, the levels of AST and ALT were followed every 3 months after surgery in 116 patients, and were divided into 2 groups at 80 IU/ml to compare the level of risk for the development of HCC. In liver biopsy tissue, group F4 (n=68/152, 45%) showed a significantly higher (P=0.0224) rate of appearance of HCC than group F3 (n=3/18, 17%). Group F4 also tended to show a higher cumulative HCC appearance rate of 55% compared with 37% for group F3 at 10 years after surgery (P=0.097). In regard to activity, the appearance rate of HCC in group A2+A3 (n=52/112, 51%) was significantly higher (P=0.0008) than that of HCC in group A1 (n=14/58, 25%). The cumulative appearance rate (60%) of HCC in group A2+A3 was significantly higher than that (31%) in group A1 at 10 years after surgery (P=0.0003). The appearance rate of HCC was significantly higher in the group (n=33/44, 75%) with a mean AST level >/=80 IU/ml than in the group (n=41/72, 57%) with a mean AST level <80 IU/ml (P=0.0496). A multivariate analysis of the risk factors for the development of HCC showed that necroinflammatory activity was a risk factor. These results suggested that the histopathologic classification (the New Inuyama Classification) of liver biopsy tissue from patients who underwent non-shunt operation for esophageal varices due to underlying cirrhosis or pre-cirrhosis is useful for predicting the development of HCC, to which the grades of necroinflammatory activity in particular are more closely related. PMID- 12644041 TI - Usefulness of viral concentration measurement by transcription-mediated amplification and hybridization protection as a prognostic factor for recurrence after resection of hepatitis B virus-related hepatocellular carcinoma. AB - We examined the usefulness of a transcription-mediated amplification and hybridization protection assay (TMA-HPA) for the evaluation of the degree of active hepatitis and estimation of recurrence after resection of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). We compared results obtained with TMA-HPA with results obtained with a branched DNA assay. Fifty-two patients with HBV-related HCC were divided into two groups; group 1 consisted of 28 patients with a high serum concentration of HBV (>/=3.7 LGE/ml) and group 2 consisted of 14 patients with a low serum concentration of HBV (<3.7 LGE/ml). Serum activities of aspartate and alanine aminotransferases were higher in group 1 than in 2 (P=0.0002, P=0.0020, respectively). The proportion of patients with mildly or moderately active hepatitis, evaluated by histologic examination, was higher in group 1 than in 2 (P=0.0025). The tumor-free survival rate was significantly lower in group 1 than in 2 (P=0.0014). In multivariate analysis, a high viral concentration (risk ratio, 6.37; 95% CI, 1.46-27.78), young age, and multiple tumors were independent risk factors for recurrence. The serum HBV concentration obtained with TMA-HPA seemed to correlate with the degree of active hepatitis more closely than that obtained with the branched DNA assay. The serum concentration of HBV, measured with TMA-HPA, is closely related to the degree of active hepatitis, and the measurement of the serum concentration of HBV is useful for estimating recurrence after resection of HBV-related HCC. PMID- 12644042 TI - Multicentric occurrence of hepatocellular carcinoma in patients with a somatic mutation of mitochondorial DNA and hepatitis C virus. AB - The relationship between the multicentric occurrence of hepatocellular carcinoma (HCC) and the frequency of mutation of mitochondrial DNA (mtDNA) in the noncancerous hepatic tissue in patients infected with hepatitis C virus was investigated. Of the 48 patients, multicentric occurrence of HCC was found in ten of 33 patients with three or more mutations in the mtDNA, whereas no patients had multicentric HCCs in 15 patients with two or fewer mutations in the mtDNA (P=0.0201). A high rate of mtDNA mutation in noncancerous hepatic tissue may be related to multicentric hepatocarcinogenesis and the hypercarcinogenic state in such patients. PMID- 12644043 TI - Alterations in the indexes of apoptosis and necrosis induced by galactosamine in the liver of Wistar rats treated with fructose-1,6-bisphosphate. AB - Galactosamine (GalN) is a hepatotoxic agent, which under determined situations provokes metabolic and energetic depletion as well as alterations in permeability, leading to cellular death. At the same time, it is known that fructose-1,6-bisphosphate (FBP) helps maintain cell energy levels and protects the cell against this lesive agent. We submitted two groups of male Wistar rats to the harmful intraperitoneal doses of GalN (400 mg/kg), one of which simultaneously received FBP (2 g/kg). Techniques were used in the analysis of the cellular components, adenosine triphosphate (ATP) and hepatic calcium and a close relationship between the types of cellular death unchained by these agents was established. The liver of the rats treated with GalN showed energy depletion and concomitant increase calcium in the hepatic tissue, which provoked higher levels of necrosis leading to reduce cellular viability. On the other hand, the group which had received GalN+FBP maintained calcium levels close to the control values and the energy rate did not decrease as much as in the GalN only group, but recovered the control values, within a period of 48 h. At the same time, the degree of apoptosis was greater than in the GalN group. This fact suggests that the FBP maintains cellular levels of ATP, thus protecting the cell from the toxic action of GalN, reducing the percentage of dead cells and causing an alteration in the pattern of the cell death, whereby there is an increase in the rate of apoptosis and a decrease in that of necrosis. PMID- 12644044 TI - Two cases of hepatic artery interruption after hepatopancreatobiliary surgery treated by prostaglandin E(1) infusion via the superior mesenteric artery. AB - In two cases of hepatic arterial flow interruption after hepatopancreatic surgery, continuous PGE(1) infusion from the superior mesenteric artery (SMA) was applied to oxygenate the liver through the portal vein. Case 1 was a 69-year-old woman with a non-functioning islet cell tumor of the pancreas. She underwent pancreatic resection following hepatic arterial infusion of anticancer drugs. Serum alanine aminotransferase (ALT) was elevated to 5500 IU/l on postoperative day (POD) 2; angiography revealed complete celiac artery obstruction. Continuous PGE(1) was administered from SMA at a rate of 0.01 &mgr;g/kg/min for 7 days. Serum ALT was normalized within 2 weeks and the peak level of serum total bilirubin (T. Bil) was 4.5 mg/dl. Case 2 was a 66-year-old man suffering from metastatic liver cancer. Complete obstruction of the proper hepatic artery was noted at the time of liver resection after hepatic arterial chemotherapy. Serum ALT was elevated to 2930 IU/l on POD 1, and PGE(1) infusion from SMA was done for the succeeding 7 days. Necrotic area was so vast that serum T. Bil rose to 19 mg/dl. However, it decreased with time. Both cases required 3 months for necrotic liver shrinkage. Doppler ultrasonography revealed that PGE(1) infusion actually increased portal blood flow. In conclusion, based on the preceding experimental backgrounds and clinical experiences, continuous PGE(1) infusion via the SMA can be a useful measure to prevent severe liver damage after hepatic arterial flow interruption through portal blood oxygenation. PMID- 12644045 TI - Effect of obstructive jaundice on the regulation of hepatic cholesterol metabolism in the rat. Disappearance of abcg5 and abcg8 mRNA after bile duct ligation. AB - The purpose of this study is to evaluate the effect of bile duct ligation (BDL) on changes of lipoprotein metabolism and hepatic gene expressions that are important for cholesterol metabolism. In male rats serum, very low-density lipoprotein cholesterol, low-density lipoprotein cholesterol and apolipoproteins B and E increased after 5 days of BDL compared with those of sham operation (control) group. Serum apolipoprotein A-IV concentration in the BDL group was lower than that in the control group. In both groups, there was no difference in hepatic lipid concentrations. Hepatic mRNA expressions of scavenger receptor B1, microsomal triglyceride transfer protein (mtp), HMG-coA reductase, cholesterol 7alpha-hydroxylase and multidrug resistance gene product 2 in the BDL group were significantly higher than that in the control group. The adipocyte determination and differentiation 1 mRNA expression in the BDL group was significantly lower than that in the control group. abcg5 and abcg8 mRNA expressions were remarkably decreased in the BDL group. In conclusion, in obstructive jaundice, metabolism of lipoprotein and proteins that are important for lipid metabolism are drastically changed probably for maintaining the hepatic lipid concentration. The remarkable down-regulation of the abcg5 and abcg8 may be an adaptive change reflecting the inability of biliary cholesterol excretion. PMID- 12644046 TI - Circulating antibody to hepatitis B core antigen does NOT always reflect the latent hepatitis B virus infection in the liver tissue. AB - Accumulating evidence suggests the presence of latent hepatitis B virus (HBV) infection in the liver of individuals negative for HBV surface antigen (HBsAg) but positive for antibodies to HBV core antigen (anti-HBc) at low titer. It remains unclear, however, whether positive anti-HBc in the serum invariably reflects latent HBV infection. In this study, we examined the presence of HBV genomes in the liver tissue of 33 donors and 30 recipients of living-related liver transplantation with positive for anti-HBc together with time course changes in their anti-HBc titer. None of these anti-HBc-positive healthy donors or recipients was positive for HBV-DNA nor anti-HBc at high titer (200 dilution) in their sera. However, HBV-DNA was detected in the liver tissue of 24 of 33 healthy anti-HBc-positive donors (72.7%) and five of 30 anti-HBc-positive recipients (16.7%). Interestingly, anti-HBc was continuously positive in all healthy donors tested. In contrast, anti-HBc titers in 75% of recipients, who were positive for anti-HBc at the time of liver transplantation, gradually decreased after the operation, and finally became negative after the mean follow up period of 9.0 months (range 1.2-45.1). Notably, HBV-DNA was never detected in the liver of those recipients who were transiently positive for anti-HBc. In conclusion, our findings suggested the possibility that presence of circulating anti-HBc does not always reflect the presence of HBV genomes in the liver tissue of anti-HBc-positive patients. PMID- 12644047 TI - Hepatitis C virus: prevalence and routes of infection among blood donors of West Mexico. AB - In Latin America few studies have explored frequency and risk factors predicting hepatitis C virus (HCV) infection in blood donors. In this study we determined the prevalence of HCV infection in blood donors from West Mexico. Potential risk factors, clinical, histological and virologic characteristics presented in this group were also evaluated. METHODS: HCV antibodies were evaluated in 57108 blood donors with commercial second-generation enzyme immunoassays. Positive results were confirmed by a recombinant immunoblot assay. Repeatedly seropositive donors were further studied for risk factors, history for hepatitis, hepatic enzymes (aspartate aminotransferase and alanine aminotransferase (AST and ALT)), liver histology and hepatitis C virus-ribonucleic acid (HCV-RNA) detection. RESULTS: A total of 499 blood donors were initially tested positive doubtful for antibodies to HCV Ab (0.8%). While there was no difference in HCV prevalence with respect to age or gender, the most frequent risk factors identified were transfusion (42%), household exposure (14.8%), multiple sex partner (6.8%) and intranasal cocaine use (2.3%). Also, we found that from a subgroup of donors tested for histological analysis, 19% presented abnormal ALT levels and 90% showed abnormal liver histology. No correlation was found between abnormal ALT levels and the presence of HCV-RNA in serum. CONCLUSION: These results demonstrate a low prevalence (0.8%) of HCV infection among Western Mexican blood donors, which was comparable to those established for Western countries, but in contrast in our study the most frequent risk factor continues being transfusion followed by household exposure. PMID- 12644048 TI - Analysis of the genotype-determining region of hepatitis A viral RNA in relation to disease severities. AB - Although hepatitis A is still a considerable problem in Japan, correlation of genomic differences of hepatitis A virus (HAV) and the clinical status of hepatitis A has not been studied. To examine whether the HAV genotype is associated with the disease severity, we analyzed the HAV genotype-determining region in sera from patients with hepatitis A of various clinical degrees. Serum samples from 47 Japanese patients with sporadic hepatitis A from 4 different areas in Japan, including 8 with fulminant hepatitis, 11 with severe acute hepatitis (AHs), and 28 with AHs, were examined for HAV RNA. The 168 base pair sequence of the genotype-determining region, which is located at the VP1/2A junction, was amplified by reverse transcription-polymerase chain reaction, and the nucleotide sequence was determined by direct sequencing. By comparing sequences for VP1/2A, we determined that 44 of 47 cases examined were of subgenotype IA, two were of subgenotype IB, and one was of subgenotype IIIA. With respect to subgenotype IA strains, individuals who were infected between 1990 and 1996 tended to show similar sequences, whereas those infected between 1997 and 1999 showed sequences slightly different from those of the 1990-1996 cases. There were no apparent associations between disease severity and sequences of the genotype-determining region. Hepatitis A of our study was caused by a variety of subgenotype IA strains, consistent with the sporadic spread of infection. Disease severity was not associated with the nucleotide sequence of the HAV genotype determining region. PMID- 12644049 TI - Simian TT virus (s-TTV) infection in patients with liver diseases. AB - Recently, we identified TTV isolates from nonhuman primates and named them simian TTV (s-TTV). To investigate the prevalence of s-TTV in humans, we examined sera from healthy individuals and patients with liver diseases in Japan for the presence of s-TTV DNA by PCR assay. s-TTV DNA was determined by nested PCR using s-TTV-specific primers designed from untranslated region of s-TTV genome. s-TTV DNA sequence was detected in three of 200 (1.5%) healthy adults but none of 48 infants without liver disease. On the other hand, s-TTV DNA was detected in 30 of 287 (10.5%) Japanese patients with liver disease. s-TTV coinfection with hepatitis B virus and hepatitis C virus were present in 16.7 and 30% of these patients, respectively, while 53.3% of patients were positive for s-TTV alone. Nucleotide sequence analyses in 20 patients confirmed that these PCR products were derived from s-TTV genome sequences and nearly 85% identical to those of s TTV prototypes from chimpanzees. Phylogenetic analysis demonstrated that all s TTV isolates from humans were distinguished clearly from the human TTV isolates. Furthermore, s-TTV in humans was classified into two different genotypes as well as simians. Our results indicate that generally 10.5% of Japanese patients with liver diseases were infected with s-TTV. The routes of s-TTV transmission from animal to human require clarification. PMID- 12644050 TI - Spontaneous regurgitation of portal blood flow normalized by meal intake in a patient with alcoholic liver cirrhosis. AB - We report a case with spontaneous regurgitation of portal blood flow (SRPBF) that was normalized by meal intake. A 41-year-old man with long-term alcohol abuse was admitted with a chief complaint of general fatigue. He was diagnosed as having alcoholic liver cirrhosis since his laboratory tests showed the abnormal liver function. Dynamic computed tomography detected numerous portosystemic shunts. Hepatic arterial portography showed the portal vein was narrow and irregular. Color Doppler imaging portrayed the direction of the blood flows in the branches of the portal vein to be retrograde. However, 30 min after meal intake on the same day, color Doppler study showed the direction of the blood flow in the first branch of right and left portal vein became normal. Color Doppler imaging is a useful technique to detect SRPBF and hemodynamic change in portal venous system after meal intake in patient under a completely physiologic condition. PMID- 12644051 TI - Expression of progenitor cell markers in livers with fulminant massive necrosis. AB - Studies of stem cells in various organs have greatly progressed, and progenitor cells have been confirmed even in liver by recognition of cytokeratin 14 (CK14), c-kit, flt-3, and CD34. We, therefore, immunohistochemically examined the expression of these progenitor cell markers in patients with confluent or massive necrosis, in addition to CK19, albumin, vimentin, and Ki-67. Our subjects were six survivors and 14 deceased patients. Expression of CK14 and c-kit was found in a small number of cells lining biliary ductule-like structures, and that of flt-3 was found in many lining cells in two deceased patients with multi-lobular necrosis. CK14 positive cells were positive for c-kit, flt-3, and CK19 in semi serial sections, but were negative for albumin, Ki-67, and CD34. In conclusion, expression of CK14 and c-kit was found in a small number of cells lining biliary ductule-like structures, and that of flt-3 was found in many cells lining biliary ductule-like structures. CK14-positive cells were positive for c-kit, but negative for CD34. Since c-kit is a hematopoietic marker, our study suggests that CK14- and c-kit-positive cells may be derived from bone marrow in liver with fulminant hepatitis. PMID- 12644052 TI - Immunohistochemical studies on the expression of P-glycoprotein and p53 in relation to histological differentiation and cell proliferation in hepatocellular carcinoma. AB - Localization of P-glycoprotein (P-gp) and p53 was immunohistochemically examined in 41 patients with hepatocellular carcinoma (HCC) in order to determine the relationship between the expression of P-gp and p53 and the degree of histological differentiation or cell proliferation in HCC. P-gp showed different patterns of expression between cancerous and cirrhotic liver hepatocytes, and the expression in cancerous tissue also varied according to the degree of histological differentiation. In cirrhotic liver hepatocytes, expression of P-gp was found on bile canalicular membranes. In the case of cancerous tissue, P-gp was localized on the canalicular membranes in well-differentiated HCC showing a trabecular pattern, as recognized cirrhotic liver hepatocytes. In moderately differentiated HCC showing pseudo-glandular patterns, predominant expression of P gp was found on the luminal side of cell membranes of the glandular ducts. The P gp expression rate was 87.5% in well-differentiated HCC, 84% in moderately differentiated HCC, and 37.5% in poorly differentiated HCC, indicating a marked decrease with decreasing degree of differentiation. On the other hand, the rate of mutation of p53, a tumor suppressor gene, was 12.5% in well-differentiated HCC, 52.0% in moderately differentiated HCC, and 85.5% in poorly differentiated HCC, showing a significant increase with decreasing degree of differentiation (P<0.005). The labeling index (LI) of proliferating cell nuclear antigen (PCNA) tended to increase with the progression of chronic liver disease, with a markedly high value of 24.0+/-1.5% in cases of HCC. The PCNA LI was 15.6+/-11.9% in well differentiated HCC, 23.1+/-15.1% in moderately differentiated HCC, and 50.1+/ 13.3% in poorly differentiated HCC, which indicated a significantly increase in poorly differentiated HCC (P<0.001). Thus, it became apparent that abnormal expressions of P-gp and p53 and the cell proliferation in HCC vary according to the degree of histological differentiation of the malignancy. This suggests that more effective chemotherapy for HCC can be potentially developed by considering the pattern and level of expression of P-gp as a mechanism of drug resistance and the extent of histological differentiation. PMID- 12644054 TI - Sequence variants of the Axin gene in hepatoblastoma. AB - The Wnt signaling pathway plays critical roles in the embryonic development and tumorigenesis. As a part of the Wnt signal transduction, the function of Axin complex is inhibited, leading to accumulation of beta-catenin. In hepatoblastomas, loss of APC (adenomatous polyposis coli) function or activation of beta-catenin that are the other two components involving in Wnt signaling has been demonstrated. Because hepatoblastoma shows immunohistochemical positivity of beta-catenin more often than its mutation frequency, we analyzed the Axin gene as a candidate to lead beta-catenin accumulation in hepatoblastoma. The coding region of the Axin gene was examined by PCR-SSCP using 24 sets of the primers in 22 hepatoblastomas and some paired normal tissues. Samples revealing aberrant band patterns were subjected to direct sequencing analysis. We identified totally eight variants in the exons and four intronic nucleotide substitutions. Seven variants in the exons were silent mutations, however, the remaining variant at codon 95 (ACG-->ATG) found in one hepatoblastoma predicted to result in an amino acid change from threonine to methionine. The paired peripheral blood DNAs from this patient also showed the same change; we suggested that it was a germline mutation of Axin gene. Our results suggest that mutation of the Axin gene may have a tumorigenic function in a subset of childhood hepatoblastomas. PMID- 12644053 TI - Meshwork pattern is an important risk factor for development of hepatocellular carcinoma in patients with HBV-related chronic hepatitis and cirrhosis. AB - We analyzed the importance of 'meshwork pattern', a sign representing severe irregularity on the intra-hepatic echogram, in hepatitis B virus (HBV)-related chronic hepatitis and cirrhosis, as a risk factor for development of hepatocellular carcinoma (HCC). Two hundred and thirty-one patients (143 men and 88 women) with HBV-related chronic hepatitis and cirrhosis who visited our hospital from January 1993 to December 1994 were enrolled in this study. Since enrollment, abdominal ultrasonography had in principle been performed every 3 months for cirrhotic patients and every 4-6 months for patients with chronic hepatitis for HCC screening. Cumulative HCC incidences in patient groups positive or negative for meshwork pattern were calculated by the Kaplan-Meier method. The incidence of HCC was significantly higher in the group positive for meshwork pattern (average incidence per year: 4.4%) than in the group negative for it (average incidence per year: 0.5%) (P<0.0001, Mantel-Cox test). On regression analysis with Cox's proportional hazards model, sex, alpha-fetoprotein and meshwork pattern were selected as independent risk factors for HCC. In conclusion, meshwork pattern appears to be an ultrasonographic sign useful for detecting a latent risk factor for HCC in patients with HBV-related chronic hepatitis and cirrhosis. PMID- 12644055 TI - Infiltrating neutrophils in bile duct-ligated livers do not promote hepatic fibrosis. AB - Bile duct obstruction causes rapid infiltration of neutrophils into the liver and leads ultimately to hepatic fibrosis. In this study, we assessed whether neutrophils play an active role in the pathogenesis of hepatic fibrosis under conditions of biliary obstruction. We performed bile duct ligation (BDL) on rats, some of which were depleted of neutrophils by means of an anti-neutrophil antiserum. Rats treated with the antiserum had 48% fewer neutrophils than control rats. Despite this, they showed no difference in either bile duct proliferation or hepatic fibrogenesis after BDL compared with control rats. In a second set of experiments, we performed BDL on mice with an underlying defect in neutrophil function due to transgenic expression of interleukin-8. Mice with neutrophil dysfunction deposited less (-22%) collagen in their livers after BDL than wild type mice, but the difference was not statistically significant. In summary, data from two independent rodent models indicate that infiltrating neutrophils do not influence hepatic fibrogenesis following bile duct obstruction. The findings suggest that neutrophils play little if any role in the immunomodulation of liver fibrosis. PMID- 12644056 TI - Bcl-2 prevents doxorubicin-induced apoptosis of human liver cancer cells. AB - We have isolated two stable human bcl-2 transfected cell lines, HCC-T-bcl and PLC bcl, that were derived from the transfection of two human hepatocellular carcinoma cell lines (HCC-T and PLC/PRF/5, respectively) with a plasmid vector containing recombinant bcl-2 (pCAGGS-bcl).) Cell lines transfected with the plasmid alone (pCAGGS-neo) were also established as controls (HCC-T-neo and PLC neo). HCC-T-neo and PLC-neo were sensitive to doxorubicin-induced apoptosis, as defined by morphological observation. Although HCC-T-neo expressed endogenous Bcl 2, the sensitivity of HCC-T-neo to doxorubicin-induced cytotoxicity was similar to that of PLC-neo, which does not express endogenous Bcl-2. In contrast, both HCC-T-bcl and PLC-bcl were more resistant to doxorubicin-induced cytotoxicity. Although these bcl-2 transfectants were resistant to the drug-induced apoptosis, Bcl-2 overexpression did not affect doxorubicin-induced growth suppression. These results suggest that the overexpression of Bcl-2 renders human HCC cells resistant to doxorubicin-induced cytotoxicity. PMID- 12644057 TI - Hepatocellular carcinoma mimicking cavernous hemangioma on angiography and contrast enhanced harmonic ultrasonography. A case report. AB - A 73-year-old man with chronic hepatitis due to hepatitis C virus was referred to our hospital for close examination of hepatic nodule. An abdominal ultrasonography revealed a mosaic pattern nodule with 3.7 cm in diameter. Arterial phase of dynamic CT revealed the small caudal part and marginal area of cranial part of the tumor were enhanced. The enhancement of marginal area of cranial part of the tumor continued up to portal phase and equilibrium phase and enhanced area was gradually filling in to the central area. On the other hand, the caudal part of the tumor was less enhanced compared with surrounding normal hepatic area in portal phase and equilibrium phase. An abdominal angiography revealed spotty tumor staining mimicking cotton-wool appearance, which is a typical finding for cavernous hemangioma. Contrast enhanced harmonic ultrasonography also showed hemangioma like finding (peripheral globular enhancing pattern). Because of these discrepancies on imagings, it was difficult to make final diagnosis of this tumor to be hepatocellular carcinoma since cavernous hemangioma cannot be completely ruled out. The pathological study of the specimen taken by US-guided percutaneous needle biopsy finally confirmed this nodule as hepatocellular carcinoma. In conclusion, we must keep in mind that some hepatocellular carcinomas could mimic hemangioma due to peliotic change or large acinar formation, therefore, needle liver biopsy may be essential for correct diagnosis if there is a discrepancy in several imaging findings. PMID- 12644058 TI - A case report: two patients with fulminant hepatitis E in Hokkaido, Japan. AB - Hepatitis E virus (HEV) has been listed as one of the major agents that cause fulminant hepatitis in HEV-endemic areas, but not in non-endemic areas. Recently, however, we experienced two cases of fulminant hepatitis E in Hokkaido, Japan: case-1 was a 34-year-old woman and case-2, a 51-year-old man, neither having a history of travelling outside Japan or contact with travellers abroad or foreigners. HEV RNA was detected in their acute-phase serum and sequencing analyses indicated that they were infected with different strains of HEV: genotype III in case-1 and IV in case-2. Case-1 patient deceased after liver transplantation, while case-2 patient survived. Based on these cases, we suggest that HEV should be considered as an aetiologic agent for fulminant hepatitis, even in non-endemic areas. PMID- 12644060 TI - Willingness of combat military personnel to donate organs. PMID- 12644061 TI - Is the motivation level of volunteers active in organ donation promotion affected by the training method? PMID- 12644062 TI - Pancreas and kidney transplantation: Tel Aviv medical center experience. PMID- 12644063 TI - Laparoscopic harvesting of kidneys for living donor kidney transplantation. PMID- 12644065 TI - The course and outcome of renal transplant recipients admitted to a general intensive care unit. PMID- 12644064 TI - BK polyoma virus nephropathy in kidney transplant recipient: the role of new immunosuppressive agents. PMID- 12644066 TI - Combination hepatitis B immune globulin and lamivudine versus hepatitis B immune globulin monotherapy in preventing recurrent hepatitis B virus infection in liver transplant recipients. PMID- 12644067 TI - Comparison of tacrolimus with cyclosporin as primary immunosuppression in patients with hepatitis C virus infection after liver transplantation. PMID- 12644068 TI - The role of nutritional support in transplant recipients: two case reports. PMID- 12644069 TI - Cholestasis and hypoalbuminemia as predictors of outcome after liver transplantation. PMID- 12644070 TI - Pushing the size limits of transplanted hearts. PMID- 12644071 TI - Parenteral fish oil administered to heart-beating organ donors and to renal transplant recipients: effects on renal function. PMID- 12644072 TI - A new immunosuppressive protocol for lung transplantation: early promising results. PMID- 12644073 TI - Tacrolimus-induced hemolytic uremic syndrome case presentation in a lung transplant recipient. PMID- 12644074 TI - Persistent high pulmonary artery pressure following orthotopic heart transplantation: inevitable poor prognosis? PMID- 12644075 TI - Outcome of unrelated surgical procedures among patients who received heart transplants. PMID- 12644076 TI - Establishment of a heart valve homograft bank using existing facilities. PMID- 12644077 TI - Israeli participation in periodical transplantation literature. PMID- 12644078 TI - Organ transplantation statistics in different countries: internet review. PMID- 12644079 TI - Preliminary results of non-cross-matched "old for old" kidney transplantation. PMID- 12644080 TI - Altruism and donation. PMID- 12644081 TI - Foreign candidates on the transplantation waiting list: an ethical dilemma. PMID- 12644082 TI - Rejection rate in living donor kidney transplantation with and without basiliximab in tacrolimus/mycophenolate mofetil-based protocol. PMID- 12644083 TI - Increased expression of P21((WAF1/CIP1)) CDKI gene in chronic allograft nephropathy correlating with the number of acute rejection episodes. PMID- 12644084 TI - The role of fiberoptic bronchoscopy evaluating transplant recipients with suspected pulmonary infections: analysis of 168 cases in a multi-organ transplantation center. PMID- 12644085 TI - Late cytomegalovirus infection occurring two or more years following liver transplantation: a report of seven cases and review of the literature. PMID- 12644086 TI - De novo tumors after liver transplantation: a single-center experience. PMID- 12644087 TI - Association of post-liver transplantation diabetes mellitus with hepatitis C virus infection. PMID- 12644088 TI - The intensive care unit course of patients undergoing liver transplantation: a report on success with a clinical pathway. PMID- 12644089 TI - Tacrolimus does not upregulate mucin gene expression after small bowel transplantation in rats. PMID- 12644090 TI - Outcome of marginal donors in heart transplantation. PMID- 12644091 TI - Lipid metabolism and immunosuppressive therapy in heart transplant recipients. PMID- 12644092 TI - Glucose metabolism in heart transplant recipients treated with FK506 or cyclosporine. PMID- 12644093 TI - Atrial fibrillation after adult lung transplantation. PMID- 12644095 TI - Impact of delayed graft function on cadaveric kidney transplant outcome. PMID- 12644096 TI - Effect of early versus delayed calcineurin inhibitor treatment on delayed graft function. PMID- 12644097 TI - Protocol for histologic evaluation of marginal grafts in kidney transplants. PMID- 12644098 TI - Cancer after renal transplantation. PMID- 12644099 TI - Posttransplant diabetes is a cardiovascular risk factor in renal transplant patients. PMID- 12644100 TI - Clinical relevance of posttransplantation HLA antibody monitoring by ELISA. PMID- 12644101 TI - Quantitation of 24-hour urine protein excretion in kidney transplant patients by the use of protein to creatinine ratio. PMID- 12644102 TI - Rapamycin use and rapid withdrawal of calcineurin inhibitors in pediatric renal transplantation. PMID- 12644103 TI - Is MELD useful in evaluating the surgical risk in liver transplantation candidates? PMID- 12644104 TI - Late mortality in patients with liver transplantation: causes and risk factors. PMID- 12644105 TI - De novo malignancies in liver transplantation. PMID- 12644106 TI - Antibodies against glutathione S-transferase T1 in patients with immune hepatitis after liver transplantation. PMID- 12644107 TI - Hepatic cirrhosis with sarcoid granulomas. Differential diagnosis and liver transplantation: a case report. PMID- 12644108 TI - Usefulness of mycophenolate mofetil in patients with chronic renal insufficiency after liver transplantation. PMID- 12644109 TI - Utility of ambulatory blood pressure monitoring for diagnosis of hypertension in liver allograft recipients. PMID- 12644110 TI - Hereditary nonsyndromic paucity of intrahepatic bile ducts as an indication for liver transplantation. PMID- 12644111 TI - Recurrence of primary biliary cirrhosis after liver transplantation. PMID- 12644112 TI - Comparison between levels of anti-HBS with a fixed administration dose of HBIG and a combination of HBIG and lamivudine for the prophylaxis of hepatitis B after liver transplantation. PMID- 12644113 TI - Comparison of monitoring trough and peak cyclosporine levels in heart transplantation. PMID- 12644114 TI - Morphologic and metabolic evaluation of the donor heart after an experimental freezing protocol. PMID- 12644115 TI - Preemptive therapy for the prevention of cytomegalovirus disease following heart transplantation directed by PP65 antigenemia. PMID- 12644116 TI - Lung transplantation in patients under mechanical ventilation. PMID- 12644117 TI - The donor lung assessment: experience of the Reina Sofia Hospital. PMID- 12644119 TI - Psychopathological features in transplant patients. PMID- 12644121 TI - Tailoring of minimal immunosuppression long term. PMID- 12644118 TI - Phase I/II trials of autologous peripheral blood stem cell transplantation in autoimmune diseases resistant to conventional therapy: preliminary results from the Spanish experience. PMID- 12644122 TI - The road toward transplantation tolerance. PMID- 12644123 TI - Swedes positive to organ donation: results from a recent opinion poll. PMID- 12644124 TI - Lack of donors limits the use of islet transplantation as treatment for diabetes. PMID- 12644125 TI - Cultural and ethical analysis of problem orientation in xenotransplantation development. PMID- 12644126 TI - Use of HTK solution in kidney preservation. PMID- 12644127 TI - Use of HTK solution for hypothermic machine perfusion: an alternative for the preservation of less than optimal donor livers?--An experimental study in rats. PMID- 12644128 TI - Creatinine clearance and viability biomarkers of machine perfused non-heart beating donors: is there a correlation? PMID- 12644129 TI - Use of thrombolytic streptokinase as a preflush in the NHBD procurement. PMID- 12644130 TI - The use of isosafe verifiable temperature control unit for liver graft storage prior to orthotopic liver transplantation. PMID- 12644131 TI - Impaired hepatic circulation despite normotension in brain-dead rats. PMID- 12644132 TI - Report from the Norwegian National Hospitals Living Donor Registry: one-year data, January 1, 2002. PMID- 12644133 TI - Laparoscopic living donor nephrectomy: introduction of simple hand-assisted technique (without handport). PMID- 12644134 TI - Hand-assisted retroperitoneoscopic living donor nephrectomy superior to laparoscopic nephrectomy. PMID- 12644135 TI - Retroaortic renal vein not a contraindication for hand-assisted retroperitoneoscopic living donor nephrectomy. PMID- 12644136 TI - Kidney transplantation in Icelandic patients 1970 to 2000. PMID- 12644137 TI - En bloc pediatric into adult recipients: the Newcastle experience. PMID- 12644138 TI - Beneficial effects of low-toxicity regimens with MMF in renal recipients with late allograft dysfunction. PMID- 12644139 TI - Conversion to sirolimus after kidney and liver transplantation: indications and outcomes. PMID- 12644140 TI - Successful transplantation of two children with antiendothelial cell antibodies. PMID- 12644141 TI - Changes in EBV gene load in renal transplant recipients. PMID- 12644142 TI - Measurement of compliance posttransplantation--the results of a 12-month study using electronic monitoring. PMID- 12644143 TI - Cardiovascular surgery in renal failure patients with or without subsequent renal transplantation. PMID- 12644144 TI - The effect of cyclosporine on homocysteine export in human cell lines. PMID- 12644145 TI - CMV increases TNF-alpha expression in a rat kidney model of chronic rejection. PMID- 12644146 TI - Expression of TGF-beta and PDGF-AA antigens and corresponding mRNAs in cytomegalovirus-infected rat kidney allografts. PMID- 12644148 TI - Survival after liver transplantation for cholangiocarcinoma has increased during the last decade. PMID- 12644147 TI - Liver transplantation for metastatic neuroendocrine tumor disease. PMID- 12644149 TI - Liver transplantation for polycystic liver disease--indications and outcome. PMID- 12644150 TI - Sirolimus in patients after liver transplantation. PMID- 12644151 TI - Combined coronary artery bypass grafting and orthotopic liver transplantation: a case report. PMID- 12644152 TI - Histologic findings associated with CMV infection in liver transplantation. PMID- 12644154 TI - Treatment with the molecular adsorbent recirculating system in patients with acute liver failure. PMID- 12644153 TI - Combination therapy of interferon alpha-2B and ribavirin for recurrent hepatitis C after liver transplantation. PMID- 12644155 TI - Graft mucosal blood flow is reduced prior to histologic evidence of rejection in mouse small bowel transplantation: a murine model. PMID- 12644157 TI - Microinvasive donor nephrectomy. PMID- 12644158 TI - Laparoscopic live donor right nephrectomy: a new technique with preservation of vascular length. PMID- 12644159 TI - Phase I study of oral bioflavonoids in cadaveric renal transplant recipients: effects on delayed graft function and calcineurin inhibitor toxicities. PMID- 12644160 TI - Clinical impact of the measurement of free radicals and nitric oxide to identify the reperfusion injury in human renal transplantation. PMID- 12644161 TI - Strategies to enhance organ viability in a non-heart-beating donor extracorporeal recirculation transplant model in pigs. PMID- 12644162 TI - How to deal with children with end-stage renal disease and severe bladder dysfunction. PMID- 12644163 TI - Anterior and posterior nutcracker syndrome: a report on 11 cases. PMID- 12644164 TI - Intermediate outcomes of dual renal allografts. PMID- 12644165 TI - Ipsilateral dual-kidney transplantation using organs declined by other centers. PMID- 12644166 TI - Renal transplantation and tuberculous renal tract disease. PMID- 12644167 TI - An evaluation of the Banff 97 classification after kidney transplantation. PMID- 12644168 TI - The impact of allograft nephrectomy on percent panel reactive antibody and clinical outcome. PMID- 12644169 TI - Male sexual dysfunction in renal transplant recipients: comparison to men awaiting transplant. PMID- 12644170 TI - Impact of pregnancy on kidney transplant. PMID- 12644171 TI - Donor bone marrow infusion in cadaveric renal transplantation. PMID- 12644173 TI - Neutralization of monokine induced by interferon-gamma during the early posttransplantation period prevents development of chronic allograft vasculopathy and graft fibrosis. PMID- 12644172 TI - Efficacy and safety of daclizumab induction for primary kidney transplant recipients in combination with tacrolimus, mycophenolate mofetil, and steroids as maintenance immunosuppression. PMID- 12644174 TI - Induced microchimerism by spleen reperfusion affords no immunological advantage in pancreas transplantation. PMID- 12644175 TI - Special issue in honor of James V. Neel. PMID- 12644177 TI - Jim Neel--some memories. PMID- 12644178 TI - Recollections of James Neel. AB - In his long and influential career Neel contributed to almost all aspects of genetic epidemiology from mutation to ethical and philosophical issues. His research spanned North America, Japan, Africa, and Latin America in a fascinating equipoise among clinical, biochemical, epidemiological, and molecular studies that have stimulated hundreds of researchers who enjoyed the controversies he generated as much as the insights he provided. Without exception, we treasure recollections of a high-principled and warm-hearted colleague whose field studies were a model for their generation. PMID- 12644179 TI - A colleague's personal account of J.V. Neel's early incorporation of isozymes and isoforms into his research on the causes and consequences of mutations. PMID- 12644181 TI - Molecular epidemiology: potential impacts on the assessment of public health. AB - In trying to decide what type of scientific paper I could prepare as a tribute to Jim Neel, I thought back over the discussions that we had over some 25 years. Sometimes these discussions were on specific topics such as how to extrapolate from mutation data in mice to those for humans following radiation or chemical exposures. On other occasions, our discussions were of a more philosophical nature, particularly on where the field of epidemiology might or needed to go. For example, what types of data are needed for assessing the public health impact of exposure to environmental agents. Perhaps because I enjoyed these discussions so much, I have chosen to take a look from a current perspective at the field of molecular epidemiology. Jim Neel would have loved to have entered into this discussion; he would have enhanced it in is own inimitable way. PMID- 12644180 TI - James Neel and the doubling dose concept. AB - The doubling dose (DD) is a very valuable concept in attempts to assess the genetic risks of radiation in man. It was long thought that the value of the doubling dose obtained from specific locus experiments in mice could be applied to man. James Neel, as a result of his studies on the offspring of atomic bomb survivors, showed that this was not so, but that different doubling doses could be inferred from different endpoints. PMID- 12644182 TI - Somatic gene mutation and human disease other than cancer. AB - While the focus of much mutation research is on germ-line mutation, somatic mutation is being found to be important in human disease. Neurofibromatosis 1 and McCune-Albright are disorders which are detected in the skin and other systems. The skin manifestations were essential for the demonstration of somatic mosaicism in neurofibromatosis 1, while analysis of blood DNA demonstrated somatic mutation in neurofibromatosis 2. Incontinentia pigmenti is also a disorder seen in skin and other tissues, but here it is the rare variant of the disorder in males, where it is usually lethal, that involves somatic mosaicism. Paroxysmal nocturnal hemoglobinuria is a disorder of the blood and cell separation of blood elements allows the demonstration of the somatic mosaicism. This review also discusses disorders in which somatic mosaicism, for mutations probably incompatible with life if the mutation had been germ-line, are likely to be involved. These include the Proteus syndrome, which involves both vascular tissues and bones, and two disorders which might be thought of as representing two subtypes of Proteus: Klippel-Trenaunay, which involves vascular tissues, and Maffuci, which involves bones. Embryonic mutations, which create mosaicism for both the soma and germ line, are being increasingly found in a number of disorders and are discussed more briefly. Finally, reverse mutations involving the soma have been recently found in several disorders and such revertant mutations are also examined. While the review focuses on the clinical importance of somatic mutations, many of the mutations found to date are tabulated. It is too early to see if there is a different pattern of somatic mutation as compared to germ-line mutation. Although the parameters to allow careful quantitation are not yet available, it seems that the frequency of gene mutation in embryonic cells is not markedly different than that in the germ-line. PMID- 12644183 TI - Biochemical and recessive visible specific locus responses of C3H/HeH to fractionated, acute radiation. AB - The recessive visible specific locus test has been widely used for many years to investigate the genetic effects of radiation in mice. We devised an electrophoretic-specific locus test so that biochemical mutations leading to alterations in the activity or amount of four enzymes and proteins, as well as charge changes could be detected. We measured the yield of recessive visible and electrophoretic mutations in the same experiment so that a direct comparison of mutation incidence could be made. Dominant visible mutations were also scored. The recessive visible specific locus response of male C3H/HeH to a fractionated dose of 3 + 3 Gy X-irradiation separated by 24 h was similar to that previously reported for the F1 hybrid widely used in mutagenesis studies, and other strains. The response of C3H/HeH was significantly greater for the recessive visible mutations than for the biochemical mutations, supporting the contention that the recessive visible loci are more mutable than others. Mutational analysis of some of the mutants showed that the lesions ranged from a very deletion (30% of chromosome 14 deleted) to a point mutation. The number of loci scored in the electrophoretic test has been reassessed, and it is now considered that six, not four were scored, and this has implications for the calculation of the doubling dose. PMID- 12644184 TI - Chlornaphazine and chlorambucil induce dominant lethal mutations in male mice. AB - Two antineoplastic agents, chlornaphazine (CN) and chlorambucil (CHL), were tested for the induction of dominant lethal mutations in male mice. Both compounds are nitrogen mustard derivatives and have been shown to be genotoxic in a variety of organisms. CN was administered intraperitoneally to DBA/2J male mice at a dosage of 0, 500, 1000, or 1500 mg/kg body weight (bw). Immediately following treatment, each male was mated at 4-day intervals to two virgin C57BL/6J females. CHL was administered intraperitoneally to C3H/HeJ and DBA/2J males at a dosage of 0, 2.5, or 5.0 mg/kg bw. These males were mated at weekly intervals to two virgin T-stock females. CN and CHL clearly induced dominant lethal mutations. CN induced dominant lethal effects in all post-meiotic germ cell stages of treated DBA males, with a clear dose-response relationship. The results with CHL-treated DBA males indicated that all post-meiotic germ-cell stages, except late-spermatids, were affected by CHL treatment, while in C3H males, CHL induced dominant lethal effects in all post-meiotic germ-cell stages. A dose-response relationship was also observed with CHL in C3H male mice. In the present experiments, regardless of the agent or the mouse strain used, spermatids appeared to be the germ-cell stage most sensitive to dominant lethal induction. PMID- 12644185 TI - Effect of smoking habit on the frequency of micronuclei in human lymphocytes: results from the Human MicroNucleus project. AB - The effect of tobacco smoking on the frequency of micronuclei (MN) in human lymphocytes has been the object of many population studies. In most reports, the results were unexpectedly negative, and in many instances smokers had lower frequencies of MN than non-smokers. A pooled re-analysis of 24 databases from the HUMN international collaborative project has been performed with the aim of understanding the impact of smoking habits on MN frequency. The complete database included 5710 subjects, with 3501 non-smokers, 1409 current smokers, and 800 former smokers, among subjects in occupational and environmental surveys. The overall result of the re-analysis confirmed the small decrease of MN frequencies in current smokers (frequency ratio (FR) = 0.97, 95% confidence interval (CI) = 0.93-1.01) and in former smokers (FR = 0.96, 95% CI = 0.91-1.01), when compared to non-smokers. MN frequency was not influenced by the number of cigarettes smoked per day among subjects occupationally exposed to genotoxic agents, whereas a typical U-shaped curve is observed for non-exposed smokers, showing a significant increase of MN frequency in individuals smoking 30 cigarettes or more per day (FR = 1.59, 95% CI = 1.35-1.88). This analysis confirmed that smokers do not experience an overall increase in MN frequency, although when the interaction with occupational exposure is taken into account, heavy smokers were the only group showing a significant increase in genotoxic damage as measured by the micronucleus assay in lymphocytes. From these results some general recommendations for the design of biomonitoring studies involving smokers can be formulated. Quantitative data about smoking habit should always be collected because, in the absence of such data, the simple comparison of smokers versus non smokers could be misleading. The sub-group of heavy smokers (> or =30 cigarettes per day) should be specifically evaluated whenever it is large enough to satisfy statistical requirements. The presence of an interaction between smoking habit and occupational exposure to genotoxic agents should be always tested. PMID- 12644186 TI - Altered gene expression in mouse livers after dichloroacetic acid exposure. AB - Dichloroacetic acid (DCA) is a major by-product of water disinfection by chlorination. Several studies have demonstrated that DCA exhibits hepatocarcinogenic effects in rodents when administered in drinking water. This chemical does not appear to be highly mutagenic, and the mechanism(s) involved in DCA induction of cancer are not clear. The present work was aimed at identifying changes in gene expression which may indicate critical alterations/pathways involved in this chemical's carcinogenic activities. We used cDNA microarray methods for analyses of gene expression in livers of mice treated with the tumorigenic dose of 2 g/l DCA in drinking water for 4 weeks. Total RNA samples obtained from livers of the control and DCA-treated mice were evaluated for gene expression patterns with Clontech Atlas Mouse 1.2 cDNA and Atlas mouse stress/toxicology arrays, and the data analyzed with AtlasImage 2.01 and one-way ANOVA in JMP4 software. From replicate experiments, we identified 24 genes with altered expression, of which 15 were confirmed by Northern blot analysis. Of the 15 genes, 14 revealed expression suppressed two- to five-fold; they included the following: MHR 23A, cytochrome P450 (CYP) 2C29, CYP 3A11, serum paraoxonase/arylesterase 1 (PON 1), liver carboxylesterase, alpha-1 antitrypsin, ER p72, glutathione S-transferase (GST) Pi 1, angiogenin, vitronectin precursor, cathepsin D (CTSD), plasminogen precursor (contains angiostatin), prothrombin precursor and integrin alpha 3 precursor (ITGA 3). An additional gene, CYP 2A4/5, had a two-fold elevation in expression. Further, in ancillary Northern analyses of total RNA isolated from DCA-induced hepatocellular carcinomas (from earlier reported studies of mice treated with 3.5 g/l DCA for 93 weeks), many of the same genes (11 of 15) noted above showed a similar alteration in expression. In summary, we have identified specific genes involved in the functional categories of cell growth, tissue remodeling, apoptosis, cancer progression and xenobiotic metabolism that have altered levels of expression following exposures to DCA. These findings serve to highlight new pathways in which to further probe DCA effects that may be critical to its tumorigenic activity. PMID- 12644187 TI - GC determination of long chain fatty acids that compose D003 in 5-mg film-coated tablets. AB - D003 is a new product which consists of a mixture of aliphatic fatty acids (C(24:0)-C(36:0)), that shows antiplatelet, antithrombotic and cholesterol lowering effects in experimental models. A gas chromatographic (GC) method using a DB-5 wide-bore column and 1-nonadecanoic acid as internal standard was developed and validated in order to determine D003 in 5 mg film-coated tablets. The acids were analyzed as methyl esters derivatives, prepared using 5% aqueous HCl-methanol. Developed method was specific for the active principle, even when samples were subjected to stress conditions. Good linearity (correlation coefficient >0.99) and accuracy (total average recovery=99.60%) were proven over a range 38-150% of the nominal concentration. Within-day and intermediate precisions at the nominal dose (100%) were <1.5%. The method was suitable for quality control and stability studies of these tablets. PMID- 12644188 TI - Chemometrics assisted spectroscopic determination of vitamin B6, vitamin B12 and dexamethasone in injectables. AB - A spectrophotometric method is described and applied to resolve ternary mixtures of the corticosteroid dexamethasone sodium phosphate and the vitamins B6 and B12. It involves multivariate calibration based on partial least-squares regression. The model was built with UV-vis absorption spectra, and was evaluated by cross validation on a number of synthetic mixtures. Satisfactory results for both artificial and commercial samples were obtained. A spectrofluorometric method was also developed for the determination of B6 in the presence of vitamin B12 and dexamethasone. The results provided by both methods for pharmaceutical formulations were compared successfully. None of the described procedures require sample pre-treatment steps. PMID- 12644189 TI - Simultaneous micellar LC determination of lidocaine and tolperisone. AB - A micellar liquid chromatography (MLC) procedure was developed for the simultaneous separation and determination of lidocaine hydrochloride (LD HCl) and tolperisone hydrochloride (TP HCl) using a short-column C18 (12.5 mm x 4.6 mm, 5 microm), sodium dodecyl sulfate (SDS) with a small amount of isopropanol, and diode array detector. The optimum conditions for the simultaneous determination of both drugs were 0.075 mol l(-1) SDS-7.5% (v/v) isopropanol with a flow rate of 0.7 ml min(-1) and detection at 210 nm. The LOD (2S/N) of LD HCl was 0.73 ng 20 microl(-1), whereas that of TP HCl was 1.43 ng 20 microl(-1). The calibration curves for LD HCl and TP HCl were linear over the ranges 0.125-500 microg ml(-1) (r(2)=0.9999) and 1.00-500 microg ml(-1) (r(2)=0.9997), respectively. The %recoveries of both drugs were in the range 98-103% and the %RSD values were less than 2. The proposed method has been successfully applied to the simultaneous determination of TP HCl and LD HCl in various pharmaceutical preparations. PMID- 12644190 TI - Improved LC method to determine ivermectin in plasma. AB - A simple, rapid and sensitive high-performance liquid chromatographic (HPLC) method has been developed to quantify Ivermectin (IVM) in plasma using an isocratic system with fluorescence detection. The method included a fast liquid phase extraction using cold methanol. HPLC separation was carried out by reversed phase chromatography with a mobile phase composed of methanol:acetonitrile:water with 0.2% acetic acid (45:50:5 v/v/v), pumped at flow rate of 2 ml min(-1). Fluorescence detection was performed at 365 nm (excitation) and 475 nm (emission). The calibration curve for IVM was linear from 0.25 to 100 ng ml(-1). The validation method yielded good results regarding linearity, precision, accuracy, specificity and recoveries. The values of the limit of detection (LOD) and limit of quantification (LOQ) were 0.032 and 0.167 ng ml(-1), respectively. PMID- 12644191 TI - A semi-automated 96-well protein precipitation method for the determination of montelukast in human plasma using high performance liquid chromatography/fluorescence detection. AB - A simple, semi-automated, protein precipitation assay for the determination of montelukast (SINGULAIR, MK-0476) in human plasma has been developed. Montelukast is a potent and selective antagonist of the cysteinyl leukotriene receptor used for the treatment of asthma. A Packard MultiPROBE II EX is used to transfer 300 microl of plasma from sample, standard, and QC sample tubes to a microtiter plate (96-well). After addition of the internal standard by a repeating pipettor, a Tomtec QUADRA 96 adds 400 microl of acetonitrile to all plasma sample wells, simultaneously, in the microtiter plate. The Tomtec is also used to transfer the acetonitrile supernatant from the plasma protein precipitation step, batchwise, to another microtiter plate for analysis by HPLC with fluorescence detection. This assay has been validated and implemented for a clinical study of over 1300 plasma samples and is comparable to manual assays in the LLOQ (lower limit of quantitation, 3 ng/ml) and in stability. This is the first semi-automated protein precipitation assay published for the analysis of montelukast in human plasma and it results in significant time savings over the manual methods, both in sample preparation and in HPLC run time. PMID- 12644192 TI - Physical and chemical evaluation of liposomes, containing artesunate. AB - As artesunate has a rapid onset of therapeutic effect and quick elimination, frequent administration is required, especially in the treatment of malaria. Such treatment courses led to bad patients' compliance, leading to high recrudescence rate. Therefore, slow release preparations seemed to be a logical approach in artesunate monotherapies, as can be developed with liposomal suspensions, especially for parenteral administration. Thus, the aim of this study was to develop sterile liposomes. The suspension was evaluated on its chemical/physical stability, including chemical degradation and crystallization of artesunate, and release capacities, by use of the dialysis technique. The maximal encapsulation degree of artesunate without crystals was 1.5 mg in 300 mg lipids per ml suspension, containing egg-phosphatidylcholine/cholesterol in a molar ratio of 4:3. The highest stability was obtained with a phosphate buffer of pH 5, which could be expected, as artesunate is almost totally encapsulated. But by reason of instability in water, the suspension containing artesunate 1 mg/ml was preferred, as the encapsulation efficiency is 100%. The in vitro release test proves that artesunate is reversibly encapsulated in liposomes. A method for sterile production of liposomes at lab-scale level is also presented. PMID- 12644193 TI - A flow injection sensor for simultaneous determination of sulfamethoxazole and trimethoprim by using Sephadex SP C-25 for continuous on-line separation and solid phase UV transduction. AB - A flow-through sensor based on integration of spectrophotometric detection and the different kinetics of retention/elution of analytes on a solid support is proposed for the simultaneous determination of sulfamethoxazole (SMZ) and trimethoprim (TMP). The solid support (Sephadex SP C-25) fills both, a microcolumn placed on-line and the sensing microzone. The intrinsic absorbance of both compounds is monitored directly on the solid phase at 269 nm and so, no derivatization step is required. Using two alternate solutions, 10(-4) M hydrochloric acid and 0.20 M NaAc/HAc (pH 5.0) buffer, the sensor responds linearly in the measuring range of 50-250 and 10-70 microg ml(-1) with detection limits of 9.5 and 0.6 microg ml(-1) (500 microl of sample volume) for SMZ and TMP, respectively. The main advantages of the sensor are simplicity, rapidity and low reagents consumption. Its application to SMZ and TMP determination in synthetic samples and pharmaceutical preparations is demonstrated. The results obtained by the proposed method were compared with those obtained by a standard HPLC method. PMID- 12644194 TI - Determination of dissociation constants of anthrocycline by capillary zone electrophoresis with amperometric detection. AB - Although daunorubicin, pharmorubicin and idarubicin are important drugs in medical and in pharmaceutical research, the dissociation constants (K(a)) of these substances are frequently not known. A method based on a non-linear model enabling the efficient determination of the K(a) of the three drugs by capillary zone electrophoresis with amperometric detection (CZE-AD) has been demonstrated. The pK(a) values obtained through the method were in agreement with the values by typical spectrophotometric method. PMID- 12644195 TI - Capillary gas chromatography determination of benzaldehyde arising from benzyl alcohol used as preservative in injectable formulations. AB - A simple, precise and accurate capillary gas-liquid chromatographic procedure has been developed to determine benzaldehyde, the toxic oxidation product of the widely used preservative and co-solvent benzyl alcohol, in injectable formulations of the non-steroidal anti-inflammatory drugs, diclofenac and piroxicam, as well as in Vitamin B-complex injection solutions. Following liquid liquid extraction with chloroform, separation and quantification are achieved on a fused silica capillary column (25 m x 0.53 mm i.d.) coated with 0.5 microm film of OV-101. 3-Chlorobenzaldehyde was used as internal standard with flame ionization as the detection mode. The ability of the system to resolve benzaldehyde peak from interfering components is good. The method displays excellent linearity over the concentration range 0.5-100 microg/ml of benzaldehyde and a precision of better than 2.5% from intra- and inter-day analyses. The quantification limit for benzaldehyde is 0.4 microg/ml. Levels of benzaldehyde in generic diclofenac and piroxicam injection formulations were found to be seven to 15 times higher than in reference formulations, and double in generic Vitamin B-complex injection formulations. PMID- 12644196 TI - Study on protolytic equilibria of lorazepam and oxazepam by UV and NMR spectroscopy. AB - Protolytic equilibria in homogeneous and heterogeneous systems of lorazepam and oxazepam, which are sparingly soluble ampholytes from the class of 1,4 benzodiazepines, were studied at 25 degrees C and ionic strength of 0.1 M. Acidity constants and equilibrium constants in a heterogeneous system were determined. On the basis of the analysis of the corresponding 13C- and 1H-NMR spectra, deprotonation site in the molecules of the investigated compounds was predicted. Finally, the correlation between chemical shifts in the 1H-NMR spectra and the acidity of the amide proton of 1,4-benzodiazepines was established. PMID- 12644197 TI - Spectrophotometric and polarographic determination of enalapril and lisinopril using 2,4-dinitrofluorobenzene. AB - The reaction of enalapril maleate and lisinopril with 2,4-dinitrofluorobenzene has been used to form colored products and polarographically active derivatives. The different experimental conditions have been optimized. The proposed methods have been validated and applied to the determination of both drugs in their commercial tablets. The results have been statistically compared with those obtained using the official HPLC methods. PMID- 12644199 TI - Detection and disposition of tolmetin in the horse. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are prohibited by the International Federation of Horse Racing Authorities but are commonly used in veterinary practice. Plasma and urinary concentrations of the NSAID tolmetin were determined by a high-performance liquid chromatographic procedure with UV detection following oral administration of a dose of 1 g to six fasted untrained standard bred mares. With a limit of quantitation (LOQ) of 0.05 microg/ml tolmetin was present in plasma for 9-12 h post-administration. Maximum concentrations of 2.1+/-0.89 microg/ml were found after 0.7+/-0.25 h. The elimination half-life was 2+/-1.25 h. Plasma protein binding at concentrations of 0.25 and 2.5 microg/ml was 92+/-4.9 and 84+/-4.2%, respectively. As early as 1 h after dosage, tolmetin could be detected in unhydrolysed urine and remained detectable up to 48 h (LOQ=0.5 microg/ml). The maximum concentrations occurred 1.8+/-0.4 h after administration. The percentage of the dose excreted as unchanged tolmetin within 12 h was 58+/-7.9%. Neither conjugates nor metabolites could be detected under the experimental conditions studied. For confirmatory analysis in doping control, an LC-MS method was developed. Analysis was performed on an ion trap LC-MS system equipped with an ESI probe in positive MS(2) mode. PMID- 12644198 TI - Structural elucidation and conformational properties of the toxin paralysin beta Ala-Tyr. AB - Larval extracts of the homotetabolous insects (i.e., Neobelleria Bullata-Insecta Diptera), cause paralysis followed by death when injected into adult flesh flies. The reason for causing these lethal effects is because the extracts contain endogenous toxins widely spread over the class of insects. Since their major effect is the paralysis they are called paralysins and are present through all the development stages. Their concentration gradually increases from larvae stage over pupation to late pharate adults indicating that paralysins have an active role in the metamorphosis. The prototype pharmacologically important dipeptide beta-alanine-tyrosine was synthesized and submitted to conformational analysis studies in hydrophilic and amphoteric environments in order to reveal the stereoelectronic properties responsible for its activity. PMID- 12644200 TI - RP-HPLC and NMR study of cis-trans isomerization of enalaprilat. AB - The angiotensin converting enzyme inhibitor, enalaprilat can exist in solution as cis and trans conformers which interconvert around the amide bond at room temperature. A HPLC with UV detection was performed to study the influence of various chromatographic operational conditions on both rotamers separation and elution of enalaprilat as a single peak. In addition nuclear overhauser enhancement difference was used for the identification of the conformers. The isomer ratio integrated from the obtained 1H NMR result were 71.5:28.5 and 76:24 at 298 and 279 K, respectively where the trans was the major form. PMID- 12644201 TI - Effects of alkali and simulated gastric and intestinal fluids on danazol stability. AB - The degradation kinetics of methanolic solution of danazol (0.020% w/v) in aqueous buffers and sodium hydroxide was investigated using stability-indicating HPLC method. The drug degrades in alkaline medium through a base-catalysed proton abstraction rather than via an oxidative mechanism involving oxygen species. The degradation followed pseudo-first-order kinetics. The rates pH-profile exhibited specific base catalysis. The stability of the drug was found to be dependent on pH, buffer concentration, buffer species (acetate, borate, phosphate) and temperature. The ionic strength did not affect the stability of the drug. The energy of activation according to Arrhenius plot was estimated to be 22.62 kcal mol(-1) at pH 12 and temperatures between 30 and 60 degrees C. The effect of simulated gastric and intestinal fluids on the drug stability was also investigated. Two major hydrolytic degradation products were separated and identified by IR, NMR and mass spectrometry and the degradative pathway suggested. PMID- 12644202 TI - Radiolysis characterization of cetostearyl alcohol by gas chromatography-mass spectrometry. AB - Methods for sample preparation, assay test and impurity test were established. Degrees of cetostearyl alcohol (CSA) radiolyses in pure state, ointment base and in chloramphenicol eye ointment (CAPEO) were determined at doses of 25 and 50 kGy. Radiolyses of CSA occur in all cases. The degrees are directly proportional to the irradiation dose in each case. Forty-two impurities and radiolysis products were identified using gas chromatography-mass spectrometry. The radiolysis products were assigned to be n-alkane, n-aldehyde and 2-methyl-1 alcohol. Accordingly, the degradation pathways of cetosteary alcohol were elucidated. Radiolysis behaviors of CSA in pure state, eye ointment base and CAPEO were studied by assay and impurity analyses. The influence of eye ointment matrixes is modest and chloramphenicol molecule exhibits slight scavenger function for cetostearyl. Both qualitative and quantitative data confirm that the radiolysis products of CSA do not cause safety concerns for human use. PMID- 12644203 TI - Spectrophotometric and fluorimetric determination of aztreonam in bulk and dosage forms. AB - Spectrophotometric and fluorimetric determination of aztreonam were achieved through its reaction with cerium (IV) in acidic medium. The spectrophotometric method involves the quantitation of the amount of ceric equivalent to aztreonam by measuring the absorbance at 317 nm and the corresponding first-derivative value at 284 nm for the blank solution against the reaction solution. Beer's law is obeyed over the concentration ranges of 1.5-4 and 1-4 microg ml(-1), respectively. Meanwhile, in the fluorimetric method, higher sensitivity was achieved by measuring the fluorescence intensity of the formed cerium (III) at lambda(em)=357 nm (lambda(ex)=257 nm) within a concentration range 150-350 ng ml( 1). Study of the reaction conditions and reaction stoichiometry were presented. Interference from L-arginine, which is frequently co-formulated with aztreonam, was tested. The proposed procedures were applied successfully to the determination of aztreonam in pure form and in presence of arginine both in laboratory mixtures and commercial vials. The proposed methods are sensitive, accurate and precise as compared with the official USP 24 HPLC method. PMID- 12644204 TI - The ICH guidance in practice: stress decomposition studies on three piperazinyl quinazoline adrenergic receptor-blocking agents and comparison of their degradation behaviour. AB - Stress degradation studies were carried out on three piperazinyl quinazoline alpha(1)-adrenergic receptor blockers, viz. prazosin, terazosin, and doxazosin, following the conditions prescribed in the parent drug stability testing guideline (Q1AR) issued by International Conference on Harmonization (ICH). All drugs showed significant decomposition at 80 degrees C in acidic conditions (0.1 M HCl) and complete degradation in alkaline conditions (0.1 M NaOH). Under both these conditions, 2-piperazinyl-6,7-dimethoxy-4-aminoquinazoline was formed as a major decomposition product in all three drugs. The degradation pattern under ICH prescribed photolytic conditions in liquid and solid states was also similar for all the drugs. The light exposure resulted in the formation of a cluster of degradation products. No degradation was observed in neutral and oxidative conditions. In solid state, all drugs were stable at 50 degrees C in a 1-month study. In alkaline conditions, the order of sensitivity to degradation of the three drugs was doxazosin>terazosin>prazosin, while the same was terazosin>doxazosin>prazosin under acidic conditions. Mechanistic explanation is provided for the variable behaviour of decomposition. PMID- 12644205 TI - Conductometric and indirect AAS determination of antimalarials. AB - Two methods are described for the determination of amodiaquine hydrochloride, chloroquine phosphate and primaquine phosphate, based on the formation of their ion-associates with [Cd(2+), Co(2+), Mn(2+) and Zn(2+)] thiocyanate, ammonium reineckate and/or sodium cobaltinitrite. The molar combining ratio reveal that (1:1) (drug:reagent) ion associates are formed for all reagents except for ammonium reineckate which form (1:2) ion associates with all studied drugs. The optimum conditions for the ion-association have been studied. Conductometric method was applied for the direct determination of the suggested drugs in bulk powders, whereas indirect atomic absorption spectrometric method, depending on the measurement of the excess metal ion present in supernatant solutions after precipitation of the ion associates is used to calculate the drug concentration. Optimum concentration ranges for the determination of aminoquinoline antimalarial drugs under consideration were 0.46-12.90 and 0.155-3.87 mg using conductometric and indirect atomic absorption spectral methods, respectively. The proposed procedures have been applied successfully to the analysis of these drugs in certain formulations and the results are favourably comparable to the official methods. PMID- 12644206 TI - The use of the new SPE methods for isolation of some tricyclic antidepressant drugs from human serum. AB - Solid-phase extraction (SPE) methods were applied to isolation of amitriptyline (AMI), imipramine (IMI) and chlorprothixene (CPX) from blood human serum. SPE was carried out using the octadecyl (C(18)) column for isolation of AMI and cyclohexyl (CH) columns in the case of IMI and CPX. The spiked serum samples were used to examine the recoveries of these drugs from C(18) and CH sorbent materials. The volume of serum sample was 500 microl. The recoveries of SPE method using CH cartridge were 100.3+/-1.63% (n=7), 99.7+/-2.3% (n=9) for IMI and CPX, respectively. The recovery of AMI from C(18) cartridge was 99.5+/-1.5% (n=8). Finally, after SPE sample clean-up step the antidepressant drugs were assayed by the own extractive-spectrophotometric methods. PMID- 12644207 TI - Spectrophotometric determination of binary mixtures of pseudoephedrine with some histamine H1-receptor antagonists using derivative ratio spectrum method. AB - A derivative spectrophotometric method is developed for the assay of three binary mixtures of pseudoephedrine with fexofenadine (mix I), cetirizine (mix II) and loratadine (mix III). The method is based on the use of the first derivative of the ratio spectrum. The ratio spectrum was obtained by dividing the absorption spectrum of the mixture by that of one of the components. The concentration of the other component was determined from its respective calibration graph treated similarly. Moreover, the influence of Deltalambda for obtaining the first derivative of the ratio spectra and the effect of the divisor concentration on the calibration graphs were studied. The described method was applied for the determination of these combinations in synthetic mixtures and dosage forms. The results obtained were accurate and precise. PMID- 12644208 TI - Voltammetric behavior of cefixime and cefpodoxime proxetil and determination in pharmaceutical formulations and urine. AB - Electrochemical reduction behavior of cephalosporins, Cefixime (CF) and Cefpodoxime Proxetil (CP) have been studied by using different voltammetric techniques in Britton-Robinson buffer system. Two well defined cathodic waves are observed for both the compounds in the entire pH range. Number of electrons transferred in the reduction process was calculated and the reduction mechanism is proposed. The results indicate that the process of both the compounds is irreversible and diffusion-controlled. The peak currents for CF and CP are found to be linear over the range of concentration 6.0 x 10(-8) to 1.2 x 10(-5) mol l( 1) and 8.8 x 10(-8) to 1.1 x 10(-5) mol l(-1), respectively. The lower detection limits are found to be 4.6 x 10(-8) and 8.52 x 10(-8) mol l(-1) for the two compounds. A differential pulse voltammetric method has been developed for the determination of these drugs in pharmaceutical formulations and urine samples. PMID- 12644209 TI - Gallamine-tetraphenylborate-modified carbon paste electrode for the potentiometric determination of gallamine triethiodide (flaxedil). AB - The construction and general performance of a novel modified carbon paste electrode for the determination of gallamine triethiodide have been developed. The electrode shows a stable, potentiometric response for gallamine in the concentration range 1 x 10(-3)-2 x 10(-6) M at 25 degrees C independent of pH in the range 5-8. The electrode passes a near-Nernstian cationic slope of 17.0+/-0.7 mV and lower detection limit of 1 x 10(-6) M with a fast response time of 20-45 s. Selectivity coefficients for gallamine relative to a number of interfering substances were investigated. There is a negligible interference from the studied cations, anions, and pharmaceutical excipients. The determination of gallamine in aqueous solution shows an average recovery of 99.5% and a mean relative standard deviation (RSD) of 1.4% at 100 microg/ml. The direct determination of gallamine in injection solution gave results that compare favorably with those obtained by the British pharmacopoeia method. Potentiometric titration of gallamine with sodium tetraphenylborate and phosphotungstic acid as a titrant has been monitored with the modified carbon paste electrode as an end-point indicator electrode. PMID- 12644210 TI - Photochemical stability of nimesulide. AB - HPLC and TLC methods for monitoring of the photochemical stability of nimesulide are presented. Solution of nimesulide sodium salt was exposed to the light of wavelengths 254 nm. The presence of degradation products (2-phenoxy4-nitroaniline and methanesulfonic acid) was observed. In the exposed sample, 2-phenoxy4 nitroaniline was detected by HPLC analysis and sulfonic acid was detected by TLC analysis. An isocratic HPLC chromatographic condition was described for determination of nimesulide in a presence of its degradation product. The sample was analysed on Separon SGX, C(18), 250 x 4.6 i.d. 7 microm analytical column. The mobile phase was consisted of a mixture of acetonitrile and ammonium phosphate (pH 7.9; 0.02 M) (35:65 v/v). UV detector was performed at 245 nm. Propylparaben was employed as an internal standard. Standard area response was linear respect to concentration of nimesulide over range 150-500 microg/ml. As a validation of the method, the accuracy and between-day precision were done. The detection limit of 2-phenoxy4-nitroaniline was 0.12 microg/ml. The solvent system for TLC analysis was consisted of ethylacetate and cyclohexane (45:55), the samples were plotted on silica gel UV-254 nm. UV lamp (254 nm) and the chemical detection were used. PMID- 12644212 TI - Expectant management with selective delayed intervention for favorable risk prostate cancer. AB - Management options for favorable risk prostate cancer are diverse, varying from a conservative approach (expectant management) to definitive treatment (radical prostatectomy or radiotherapy.) Several studies have suggested that expectant management provides similar 10-year survival rates and quality-adjusted life years compared with radical prostatectomy or radiotherapy. Expectant management alone clearly deprives some patients with potentially curable life-threatening disease of the opportunity for curative therapy. However, every series of conservative management contains a substantial subset of long-term survivors, particularly in the group with favorable clinical parameters. We have conducted a clinical study to evaluate a novel approach in which the choice between a definitive therapy and conservative policy is determined by the rate of PSA increase or the development of early, rapid clinical and/or histologic progression. This strategy, which has never been previously evaluated, offers the powerful attraction of individualizing therapy according to the biological behavior of the cancer. This would mean that patients with slowly growing malignancy would be spared the side effects of radical treatment, while those with more rapidly progressive cancer would still benefit from curative therapy. Doubling time varied widely. In this series of 200 patients, neither grade, stage, nor baseline PSA predicted the PSA doubling time. Thirty-three percent of patients had a PSA doubling time (T(D)) > 10 years. Doubling time appears to be a useful tool to guide treatment intervention for patients managed initially with expectant management. A doubling time of less than 2 years appears to identify patients at high risk for local progression in spite of otherwise favorable prognostic factors. Fifteen to 20% of patients will fall into this category. The remainder have a high chance of remaining free of recurrence and progression for many years. Watchful waiting is clearly appropriate for patients who are elderly, have significant co-morbidity, and have favorable clinical parameters. The use of co-morbidity indices like the Index of Co-existent Disease (ICED) facilitate the identification of patients whose life-expectancy is diminished relative to the natural history of their prostate cancer. The likelihood of a prostate cancer death in these patients is low. Furthermore, many healthy patients fall into a grey zone where the benefits of treatment are unclear. In these patients, a policy of close monitoring with selective intervention for the 15-20% who progress rapidly is appealing. This approach is currently the focus of several clinical trials. PMID- 12644213 TI - Primary high grade malignant lymphoma of bladder. AB - Primary malignant lymphoma of bladder is one of the rarest extranodal site for lymphomas. Less than 100 cases have been reported so far. A history of chronic cystitis has been shown to be a preceding feature in many cases (40%) of primary bladder lymphoma. Most of the cases reported in the literature have a low-grade lymphoma including the subtypes of mucosa-associated lymphoid tissue. The diffuse large cell lymphoma is the most frequent type among the subtypes of high-grade bladder lymphomas. In this report, a case with high-grade primary malignant lymphoma of bladder is reported and the clinical, pathological aspects of diseases are reviewed. PMID- 12644214 TI - DD23 Biomarker: a prospective clinical assessment in routine urinary cytology specimens from patients being monitored for TCC. AB - BACKGROUND: A prospective clinical study was conducted to assess the ability of the DD23 murine monoclonal antibody to enhance detection of bladder cancer in routine alcohol fixed urine cytology samples. METHODS: Prospectively, 308 bladder cytology specimens were obtained from patients with a history of bladder cancer with a mean age of 71.4+/-11.9 (27% female, 73% male). Data included 121 biopsy confirmed results and 187 cystoscopy results to assess presence or absence of cancer. Thirty-five normal cytology specimens were obtained from asymptomatic men and women between 55-85 years of age. Separate slides from the alcohol fixed cytology specimens were stained using the Papanicolaou (Pap) and Feulgen staining procedures. The DD23 assay was performed using an avidin-biotin alkaline phosphatase immunocytochemical procedure, with a single urothelial cell exhibiting intense immunostaining sufficient to make a positive call. RESULTS: Pap-Feulgen cytopathology for the 308 cases yielded an overall sensitivity of 65.5% and a specificity of 85.1%, and the DD23 biomarker alone yielded a sensitivity of 80.5% and a specificity of 59.7%. Analysis of the voided urines only (n=164) yielded sensitivities of 61.0% and 73.2% and specificities of 86.2% and 67.5% for cytopathology and DD23 alone, respectively. Results in 49 bladder wash urine cytology cases produced a sensitivity of 70.2% and 100% and specificities of 92.3% and 61.5% for cytopathology and DD23 alone, respectively. In 133 patients that underwent biopsy or had positive cystoscopy results, cytopathology yielded a sensitivity of 65.5% and a specificity of 69.6% while DD23 yielded a sensitivity of 80.5% and a specificity of 58.7%. In 25 biopsy confirmed low-grade cancers, DD23 improved cancer detection from 32% to 72% when compared to cytopathology. The DD23 biomarker had a specificity of 85.7% in 35 age-matched normal asymptomatic control specimens. CONCLUSIONS: The DD23 biomarker is an adjuvant test that provides improved detection of bladder cancer in cytology specimens and enhances the sensitivity of the cytopathology diagnosis, especially in low-grade cancers. PMID- 12644215 TI - Sural nerve graft during laparoscopic radical prostatectomy. Initial experience. AB - OBJECTIVE: To determine the technical feasibility of sural nerve grafting for restoration of cavernous nerve continuity after radical retropubic prostatectomy by the laparoscopic approach. MATERIALS AND METHODS: 15 potent men (mean age: 56+/-4 years) underwent laparoscopic radical prostatectomy with deliberate wide uni- or bilateral neurovascular bundle resection. After prostatectomy, but before the vesicourethral anastomosis, an autologous sural nerve graft was interposed between the divided end of the cavernous nerves using a laparoscopic approach. RESULTS: All 15 procedures could be completed laparoscopically. Visual cooptation between the nerve graft and the cut ends of the neurovascular bundles was possible in all 15 cases. Surgical time for the entire procedure ranged between 210 to 275 min with 30 to 60 min of this time required for the nerve harvest and graft. CONCLUSION: This experience demonstrates that sural nerve graft replacement is technically feasible by the laparoscopic approach. The magnified optics, bloodless surgical field, and improved instrumentation create an optimal environment for sural nerve grafting to the cut ends of the neurovascular bundles. Quality of life survey data at 12 to 18 month follow-up will be needed to assess functional return. PMID- 12644216 TI - The free/total prostate-specific antigen ratio (%fPSA) is the best predictor of tumor involvement in the radical prostatectomy specimen among men with an elevated PSA. AB - BACKGROUND: The relationship between the free-to-total prostate-specific antigen ratio (%fPSA) and prostate cancer (CaP) pathology remains controversial. Previous reports have shown a direct correlation between %fPSA and prostate volume as well as an indirect correlation between %fPSA and unfavorable CaP pathology, particularly among men with an elevated PSA. We evaluated the use of %fPSA to predict CaP pathology including percent of tumor involvement in the radical prostatectomy (RP) specimen. METHODS: We prospectively analyzed 124 consecutive patients with CaP who underwent RP. In all patients, preoperative frozen serum was analyzed for assessment of %fPSA (Abbott Axsym). Pathologic review was performed using whole mount sections and total tumor volume was determined by planimetry. Statistical comparison between %fPSA and pathology was performed using log transformation. RESULTS: Percent fPSA was indirectly correlated with prostate volume in both the entire group (N=124) and among those patients (N=87) with a total PSA >4 ng/mL (P<0.001). Overall, both %fPSA and total PSA also correlated with total tumor volume (P=0.03 and P=0.01, respectively) and Gleason sum (P<0.001 and P<0.01). When we evaluated the percent of tumor involvement (tumor density) defined as the volume of tumor per gland divided by total gland volume, for the entire population, both total PSA and %fPSA were predictive with equal significance (P<0.001). However, among the subset of patients with a PSA>4.0 ng/mL, there was only a significant correlation between tumor density and %fPSA as compared to total PSA (P<0.001 vs. P=0.06, respectively). CONCLUSIONS: Independent of prostate volume, %fPSA is reflective of CaP pathology. Specifically, %fPSA was inversely correlated with tumor volume, Gleason sum and ECE. Among patients with modest PSA elevations, %fPSA was better than PSA in predicting percent of tumor involvement (tumor density) in the RP specimen. PMID- 12644217 TI - Evaluation of PCR-ELISA for determination of telomerase activity in prostate needle biopsy and prostatic fluid specimens. AB - The conventional TRAP assay will determine telomerase activity in tissue or other specimens. However, methodological disadvantages limit its clinical use. We evaluated a modified TRAP assay, the telomerase PCR-ELISA, as a practical clinical system for measuring its activity in conjunction with prostate cancer (PCa). We examined telomerase activity by both TRAP and PCR-ELISA assays in 48 sextant needle biopsy (SNB) specimens from dye-marked areas of the prostate glands of 7 PCa patients. Each specimen was histologically confirmed as cancerous or cancer-free by examining a paired specimen taken from the same marked area. In addition, prostatic fluid (PF) specimens were analyzed from 18 patients, 9 of whom were diagnosed with PCa while 9 were diagnosed as cancer-free but mostly with BPH. The results on individual SNB specimens matched well for the two methods. The sensitivity (91%) and specificity (69%) for the PCR-ELISA measurements were consistent with those for the conventional TRAP assay, 88% and 81%, respectively. Quantitatively, with the PCR-ELISA assay, the mean telomerase activity (24.5+/-28.4 units) per needle core with PCa cells was significantly higher than that in needle cores without PCa cells (7.2+/-2.2 unit), as it was with the conventional TRAP assay, namely 25.6+/-27.8 units and 7.3+/-1.8 units, respectively. In PF specimens from PCa patients, which had a lower mean telomerase than was found in needle cores containing PCa cells (7.1+/-1.5 units in the PCR-ELISA, 7.2+/-1.8 units in the conventional TRAP assay), statistical analysis showed good matching between the results from the two assays, overall. In conclusion, the PCR-ELISA can be considered a reliable method to determine telomerase activity as an adjunct in the diagnosis and treatment of prostate cancer. PMID- 12644219 TI - Incidental detection of population-based prostate cancer incidence rates through transurethral resection of the prostate. AB - OBJECTIVE: To provide an updated assessment of the influence transurethral resection of the prostate (TURP) has on incidental detection of prostate cancer and to characterize stage, histologic grade, and treatment patterns among these cases. METHODS: Analyses were based on 17,310 histologically confirmed prostate cancer cases ages 45 years and older recorded in the Utah Cancer Registry between 1980 and 1999 and 6426 TURP procedures recorded in the Utah Hospital Discharge Database from 1992 through 1999. An algorithm was developed for identifying TURP detected prostate cancer incidence. RESULTS: Age-specific TURP-detected prostate cancer incidence rates tended to be flat between 1980 and 1990, decline through 1994, and then level off. Much of the decrease corresponds to the prostate specific antigen (PSA) screening induced peak and subsequent fall in total prostate cancer incidence rates. Leveling off in the TURP-detected rates between 1994 and 1999 corresponds with a leveling off in the total prostate cancer incidence rates. The percentage of prostate cancer detected by TURP significantly increases with age, within each age group, but decreases over calendar years. For ages 45 years and older, the percentage of TURP-detected cases was 39.0% in 1980 to 1984, 33.9% in 1985 to 1989, 12.2% in 1990 to 1994, and 7.4% in 1995 to 1999. TURP-detected cases were significantly less likely to undergo radical prostatectomy or radiation therapy across age groups, despite having lower stage and histologic grade at diagnosis. CONCLUSION: TURP-detected prostate cancer rates have leveled off in the latter part of the 1990s, explaining about 10% of all detected cases. These cases have relatively good biologic potential and are less likely to pursue aggressive therapy. PMID- 12644218 TI - Monitoring gene expression profile changes in bladder transitional cell carcinoma using cDNA microarray. AB - PURPOSE: Differential gene expression profiles between normal bladder mucosas and bladder transitional cell carcinomas TCC were detected. MATERIALS AND METHODS: cDNA microarrays were prepared by spotting PCR products of 12,800 human genes onto specially treated glass slides. The cDNA probes were prepared by labeling normal bladder mucosa mRNA and TCC tissue mRNA with Cy3-dUTP and Cy5-dUTP respectively through reverse transcription. The arrays were then hybridized against the cDNA probe mixture and the fluorescent signals were scanned. The ratios of Cy5/Cy3 were computed. Northern analysis was used to confirm the results of microarray hybridization. RESULTS: Eighty-three genes (0.65%), whose ratios of Cy5/Cy3 were greater than 4.0 or less than 0.25, were screened out after 10 groups of hybridization. In the cancerous tissues 28 of them showed higher expression and 55 lower. Twenty-three genes are unregistered in GenBank. These differentially expressed genes are always involved in the physiological processes such as signal transduction, apoptosis and cell cycle, etc. CONCLUSIONS: This technique provides a powerful method for quantitative analysis of the expression levels of thousands of genes in parallel, and is used to identify genes involved in TCC carcinogenesis. The data obtained by this means are comparable to those obtained by other methods. Using cDNA microarrays to define alterations in gene expression associated with a specific cancer may be an efficient way to uncover the clues to specific molecular derangements that account for its pathogenesis and thus identify potential targets for therapeutic intervention. PMID- 12644221 TI - Multi locus fingerprinting of Listeria monocytogenes by sequence-specific labeling of DNA probes combined with array hybridization. AB - We have developed an alternative multi locus sequence typing (MLST) approach that targets the variable genetic changes directly in a DNA array format. Our approach is based on DNA array hybridization in combination with sequence-specific labeling of oligonucleotide probes. Listeria monocytogenes was chosen for the development and evaluation of the assay. The genes hlyA, iap, flaA, inlA and actA were targeted. Twenty-nine suitable probe regions were identified within these genes. The DNA array results from 32 different strains were compared to serotype and amplified fragment length polymorphism data. This comparison showed that our DNA array method gave good discrimination between the strains analyzed. In conclusion, the DNA array-based MLST method is a promising tool for fingerprint bacteria. PMID- 12644220 TI - Role of oxidative stress and mitochondrial changes in cyanobacteria-induced apoptosis and hepatotoxicity. AB - Microcystins produced by cyanobacteria are potent and specific hepatotoxins; however, the mechanisms of microcystin-induced hepatotoxicity have not been fully elucidated. The induction of free radical formation and mitochondrial alterations are two major events found in microcystin-treated cultured rat hepatocytes. The mitochondrial alterations, i.e. loss of mitochondrial membrane potential and mitochondria permeability transition are now recognized as key steps in apoptosis. The activation of calpain and Ca(2+)/calmodulin-dependent protein kinase II is believed to be critical in the microcystin-induced apoptotic process. PMID- 12644222 TI - Tylosin-resistant bacteria cultivated from agricultural soil. AB - In this study we analyzed the numbers and types of cultivable tylosin-resistant bacteria from six agricultural soils that differed with respect to their association with subtherapeutic antibiotic use. The proportion of tylosin resistant bacteria to the total number of bacteria cultivated was significantly higher (7.2-16.5%) at three sites affected by subtherapeutic antibiotic use compared to three sites unaffected by subtherapeutic antibiotic use (0.7-2.5%). We also detected differences in the types of cultivable tylosin-resistant bacteria. At a site affected by subtherapeutic antibiotic use, we detected tylosin-resistant bacteria from the alpha- and beta-subdivisions of Proteobacteria. In contrast, at a site unaffected by subtherapeutic use, we detected only Streptomyces-like (high-G+C Gram-positive) tylosin-resistant bacteria. Our results suggest a link between subtherapeutic use of antibiotics and the numbers and types of antibiotic-resistant bacteria in nearby soil. However, other factors, such as soil type and temporal variation, may have also contributed to the differences observed. PMID- 12644223 TI - Identification and characterization of a new exopolysaccharide biosynthesis gene cluster from Streptomyces. AB - We report the identification and characterization of the ste (Streptomyces eps) gene cluster of Streptomyces sp. 139 required for exopolysaccharide (EPS) biosynthesis. This report is the first genetic work on polysaccharide production in Streptomyces. To investigate the gene cluster involved in exopolysaccharide 139A biosynthesis, degenerate primers were designed to polymerase chain reaction amplify an internal fragment of the priming glycosyltransferase gene that catalyzes the first step in exopolysaccharide biosynthesis. Screening of a genomic library of Streptomyces sp. 139 with this polymerase chain reaction product as probe allowed the isolation of a ste gene cluster containing 22 open reading frames similar to polysaccharide biosynthesis genes of other bacterial species. Involvement of the ste gene cluster in exopolysaccharide biosynthesis was confirmed by disrupting the priming glycosyltransferase gene in Streptomyces sp. 139 to generate non-exopolysaccharide-producing mutants. PMID- 12644224 TI - The effect of soil: water ratios on the mineralisation of phenanthrene: LNAPL mixtures in soil. AB - Contamination of soil by polycyclic aromatic hydrocarbons is frequently associated with non-aqueous-phase liquids. Measurement of the catabolic potential of a soil or determination of the biodegradable fraction of a contaminant can be done using a slurried soil respirometric system. This work assessed the impact of increasing the concentration of transformer oil and soil:water ratio on the microbial catabolism of [(14)C]phenanthrene to (14)CO(2) by a phenanthrene degrading inoculum. Slurrying (1:1, 1:2, 1:3 and 1:5 soil:water ratios) consistently resulted in statistically higher rates and extents of mineralisation than the non-slurried system (2:1 soil:water ratio; P<0.01). The maximum extents of mineralisation observed occurred in the 1:2-1:5 soil:water ratio microcosms irrespective of transformer oil concentration. Transformer oil concentrations investigated displayed no statistically significant effect on total mineralisation (P>0.05). Soil slurries 1:2 or greater, but less than 1:5 (soil:water), are recommended for bioassay determinations of total contaminant bioavailability due to greater overall mineralisation and improved reproducibility. PMID- 12644225 TI - A tripartite microbial reporter gene system for real-time assays of soil nutrient status. AB - Plant-derived carbon is the substrate which drives the rate of microbial assimilation and turnover of nutrients, in particular N and P, within the rhizosphere. To develop a better understanding of rhizosphere dynamics, a tripartite reporter gene system has been developed. We used three lux-marked Pseudomonas fluorescens strains to report on soil (1) assimilable carbon, (2) N status, and (3) P-status. In vivo studies using soil water, spiked with C, N and P to simulate rhizosphere conditions, showed that the tripartite reporter system can provide real-time assessment of carbon and nutrient status. Good quantitative agreement for bioluminescence output between reference material and soil water samples was found for the C and P reporters. With regard to soil nitrate, the minimum bioavailable concentration was found to be greater than that analytically detectable in soil water. This is the first time that bioavailable soil C, N and P have been quantified using a tripartite reporter gene system. PMID- 12644226 TI - Actinobacillus pleuropneumoniae serotype 7 siderophore receptor FhuA is not required for virulence. AB - A ferrichrome receptor, FhuA, was identified in Actinobacillus pleuropneumoniae serotype 7. An isogenic mutant with a deletion in the ferrichrome uptake receptor gene (fhuA) was constructed and examined in an aerosol infection model. The disease caused by the mutant was indistinguishable from disease induced by A. pleuropneumoniae serotype 7 wild-type; an isogenic mutant lacking expression of the exbB gene that is required for the uptake of transferrin-bound iron retained the ability to utilize ferrichrome, thereby indicating that an energy-coupling mechanism involved in ferrichrome transport remains to be identified. PMID- 12644227 TI - Degradation of vanillic acid and production of guaiacol by microorganisms isolated from cork samples. AB - The presence of guaiacol in cork stoppers is responsible for some cases of cork taint causing unpleasant alterations to wine. We have performed a characterization of the cork-associated microbiota by isolating 55 different microorganisms: eight yeast, 14 filamentous fungi or molds, 13 actinomycetes and 20 non-filamentous bacteria. A screening for degradation of vanillic acid and guaiacol production showed that none of the filamentous fungi could achieve any of these processes. By contrast, five of the eight yeast strains isolated were able to degrade vanillic acid, although it was not converted to guaiacol. Guaiacol production was only detected in four bacterial strains: one isolate of Bacillus subtilis and three actinomycetes, Streptomyces sp. A3, Streptomyces sp. A5 and Streptomyces sp. A13, were able to accumulate this compound in both liquid media and cultures over cork. These results suggest that guaiacol-mediated cork taint should be attributed to the degradative action of vanillic acid by bacterial strains growing on cork. PMID- 12644229 TI - Novel pyrazine metabolites found in polymyxin biosynthesis by Paenibacillus polymyxa. AB - A complex mixture of methyl-branched alkyl-substituted pyrazines was found in the growth medium of the polymyxin-producing bacterium Paenibacillus polymyxa, and of these, seven are new natural compounds. A total of 19 pyrazine metabolites were identified. The dominant metabolite was 2,5-diisopropylpyrazine as identified using a combination of high-resolution mass spectrometry, (1)H- and (13)C-nuclear magnetic resonance, gas chromatography-mass spectrometry as well as co-elution with an authentic standard. Its biosynthesis was correlated with growth and production was strongly stimulated by valine supplementation. The other pyrazine metabolites, all related pyrazines with either one, two or three alkyl substituents, were identified by means of their mass spectral data and/or co elution with authentic standards. PMID- 12644228 TI - HIV Tat, its TARgets and the control of viral gene expression. AB - The human immunodeficiency virus (HIV-1) (transactivator of transcription (Tat)) protein is a pleiotropic factor that induces a broad range of biological effects in numerous cell types. At the HIV promoter, Tat is a powerful transactivator of gene expression, which acts by both inducing chromatin remodeling and by recruiting elongation-competent transcriptional complexes onto the viral LTR. Besides these transcriptional activities, Tat is released outside the cells and interacts with different cell membrane-associated receptors. Finally, extracellular Tat can be internalized by cells through an active endocytosis process. Here we discuss some of the molecular mechanisms involved in intracellular and extracellular Tat function. PMID- 12644230 TI - A novel computerised image analysis method for the measurement of production of conidia from the aphid pathogenic fungus Erynia neoaphidis. AB - A semi-automated method has been developed for the quantification and measurement of conidia discharged by the aphid pathogen Erynia neoaphidis. This was used to compare conidiation by E. neoaphidis-mycosed pea aphid cadavers, mycelial plugs cut from agar plates, mycelial pellets from shake flasks and by mycelial pellets from different phases of liquid batch fermenter culture. Aphid cadavers discharged significantly more and significantly smaller conidia than plugs or pellets. The volume of conidia discharged was stable over the period of discharge (80 h), but more detailed analysis of the size frequency distribution showed that more very small and very large conidia were discharged after 5 h incubation than after 75 h incubation. Biomass harvested at the end of the exponential growth phase in batch fermenter culture produced significantly more conidia than biomass from any other growth phase. The implications of these findings for the development of production and formulation processes for E. neoaphidis as a biological control agent are discussed. PMID- 12644231 TI - Truffle thio-flavours reversibly inhibit truffle tyrosinase. AB - Tyrosinase is an enzyme having two copper atoms at the reactive site occurring in prokaryotic and eukaryotic organisms. In animals tyrosinase is responsible for pigmentation, in plants for protection of injured tissues or, as in fungi, to harden cell walls. Some of us have previously shown that tyrosinase is involved in truffle development and differentiation. Here we present the purification, the molecular properties and the reversible inhibition of Tuber melanosporum tyrosinase by dimethyl-sulfide and bis[methylthio]methane, the main flavour compounds of black and whitish truffles. The MW(r) is 39000. L-3,4 dihydroxyphenylalanine and L-tyrosine stain corresponding bands as expected for a true tyrosinase. Phenylthiourea, diethyldithiocarbamate and mimosine inhibit L tyrosine and L-3,4-dihydroxyphenylalanine oxidation. PMID- 12644232 TI - Linear beta-mannose-containing polysaccharide, beta-xylan, and amylose from the cultured photobiont Trebouxia sp. of the ascolichen Ramalina celastri. AB - The cultured photobiont Trebouxia sp. of Ramalina celastri was successively extracted at 100 degrees C with hot water, 2% aqueous KOH, and 10% aqueous KOH to give polysaccharide-containing fractions A (2.9%), B (3.9%), and C (0.9% yield) respectively. The intact biont contained 3.8% amylose, which was present in each fraction, and was identified by a blue color formed with iodine solution. In fraction A, and following retrogradation from aqueous solution, it was characterized by (13)C-NMR spectroscopy. Fraction B was treated with alpha amylase to give a water-soluble fraction consisting mainly of beta-mannose containing polysaccharides (1.5% yield), whose main component had dn/dc 0.162 and M(r) 17 kDa. Fraction C was subjected to freeze-thawing and the precipitate was treated with alpha-amylase to give a resistant, linear, low molecular mass (1- >4)-linked beta-xylan. The beta-D-mannopyranan preparation contained mainly of 3 O- (28%), 4-O- (11%), and 6-O-substituted Manp units (35%), with 3-O-substituted Rhap units (11%). A controlled Smith degradation provided a beta-mannan with nonreducing end- (8%), 3-O- (85%) and 6-O-substituted units, showing (1-->3)- and (1-->6)-linked structures in the original polysaccharide. These could be present as block-type structures. PMID- 12644233 TI - Transferrin-dependent expression of TbpA by Histophilus ovis involves a poly G tract within tbpA. AB - A poly G tract in tbpA of Histophilus ovis strain 3384Y was suspected of being responsible for the transferrin (Tf)-dependent expression of TbpA. The region encompassing the poly G tract was amplified using DNA from H. ovis strains 9L and 3384Y grown under iron-replete conditions and under iron-restricted conditions in the presence of bovine Tf. Sequence analysis of the amplification products revealed that regardless of the growth conditions, the poly G tract in strain 9L contained eight Gs, a situation that maintains the correct reading frame of the gene. Similarly, the poly G tract in strain 3384Y contained eight Gs when the organisms were grown under iron-restricted conditions in the presence of bovine Tf but when grown under iron-replete conditions, the poly G tract contained nine Gs resulting in a frame shift and the introduction of a premature stop codon. It is concluded that the Tf-dependent expression of TbpA in H. ovis strain 3384Y is due to a form of phase variation. PMID- 12644234 TI - Electron transfer from Shewanella algae BrY to hydrous ferric oxide is mediated by cell-associated melanin. AB - Shewanella algae BrY uses insoluble mineral oxides as terminal electron acceptors, but the mechanism of electron transfer from cell surface to mineral surface is not well understood. We tested the hypothesis that cell-associated melanin produced by S. algae BrY serves as an electron conduit for bacterial mineral reduction. Results from Fourier transform infrared spectroscopy and cell surface hydrophobicity assays indicated that extracellular melanin was associated with the cell surface. With H(2) as electron donor, washed cell suspensions of melanin-coated S. algae BrY reduced hydrous ferric oxide (HFO) 10 times faster than cells without melanin. The addition of melanin (20 microg ml(-1)) to these melanin-free cells increased their HFO reduction rate two-fold. These results suggest that cell-associated melanin acts as an electron conduit for iron mineral reduction by S. algae BrY. PMID- 12644235 TI - CopZ from Bacillus subtilis interacts in vivo with a copper exporting CPx-type ATPase CopA. AB - The structure of the hypothetical copper-metallochaperone CopZ from Bacillus subtilis and its predicted partner CopA have been studied but their respective contributions to copper export, -import, -sequestration and -supply are unknown. DeltacopA was hypersensitive to copper and contained more copper atoms cell(-1) than wild-type. Expression from the copA operator-promoter increased in elevated copper (not other metals), consistent with a role in copper export. A bacterial two-hybrid assay revealed in vivo interaction between CopZ and the N-terminal domain of CopA but not that of a related transporter, YvgW, involved in cadmium resistance. Activity of copper-requiring cytochrome caa(3) oxidase was retained in deltacopZ and deltacopA. DeltacopZ was only slightly copper-hypersensitive but deltacopZ/deltacopA was more sensitive than deltacopA, implying some action of CopZ that is independent of CopA. Significantly, deltacopZ contained fewer copper atoms cell(-1) than wild-type under these conditions. CopZ makes a net contribution to copper sequestration and/or recycling exceeding any donation to CopA for export. PMID- 12644236 TI - Identification and characterization of four proteases produced by Streptococcus suis. AB - Streptococcus suis is an important worldwide swine pathogen. In this study, we investigated the production of proteases by S. suis serotype 2. Proteases were identified and characterized using chromogenic and fluorogenic assays and zymography. An Arg-aminopeptidase with a molecular mass of 55 kDa was found to be both cell-associated and extracellular. Cell-associated chymotrypsin-like and caseinase activities, belonging to the serine- and metalloprotease classes respectively, were also detected. Lastly, a dipeptidyl peptidase IV (DPP IV) with a molecular mass of 70 kDa was detected in both whole cells and culture supernatants of S. suis serotype 2. Arg-aminopeptidase, caseinase and DPP IV activities were detected in all strains of S. suis serotype 2 tested whereas the chymotrypsin-like activity was only detected in European virulent strains of serotype 2. The optimum pH for all four proteases was between 6 and 8, and the optimum temperature ranged from 25 to 42 degrees C. This is the first report on the production of proteases by S. suis. Further investigations will determine the possible contribution of these proteases in the pathogenicity of S. suis serotype 2. PMID- 12644237 TI - Bacteriocin-like substance (BLS) production in Aeromonas hydrophila water isolates. AB - 30 Aeromonas hydrophila water isolates were tested for bacteriocin-like substance (BLS) production using a target panel of closely related microorganisms and other Gram-positive and Gram-negative bacteria, including food-borne pathogens. A. hydrophila showed antibacterial activity against one or more indicator microorganisms, but the activity emerged only with non-phylogenetically related genera or species. In particular all A. hydrophila showed antibacterial activity against one or more of the tested Staphylococcus strains, five against Listeria spp. (Listeria seeligeri, Listeria welshimeri and Listeria ivanovii), and eight presented a weak antagonistic activity towards Streptococcus agalactiae and Lactobacillus spp. Inhibitory activity was not observed against the other Gram positive (Listeria monocytogenes, Listeria innocua and Enterococcus spp.) and Gram-negative tested strains, including Aeromonas sobria, Aeromonas caviae and the same A. hydrophila, when used as indicator. Anti-staphylococcal activity was observed with a gradual increase of the inhibition zone during incubation and seemed to be influenced by A. hydrophila hemolytic expression. Extrachromosomal analysis showed the presence, in 70% of the strains, of one to five plasmids with molecular masses ranging from 2.1 to 41.5 MDa, but it was not possible to relate this result with BLS production. PMID- 12644238 TI - Isolation and characterisation of a novel bacteriocin produced by Bacillus thuringiensis strain B439. AB - Bacillus thuringiensis strain B439 produces a bacteriocin-like inhibitory substance in its growth medium. This antimicrobial peptide, referred to as thuricin 439, acts as a bacteriocidal peptide and exhibits an apparent narrow range of inhibitory activity, essentially only affecting growth of Bacillus cereus and B. thuringiensis strains. It remains active over a relatively wide pH and temperature range, showing no loss of activity following heat treatments up to 80 degrees C. Purification of thuricin 439 was achieved using several chromatographic steps, which resulted in the identification of two peptides with inhibitory activity. These two peptides were shown to possess identical N terminal sequences, but different molecular masses. PMID- 12644239 TI - Coaggregation between freshwater bacteria within biofilm and planktonic communities. AB - The coaggregation ability of bacteria isolated from a freshwater biofilm was compared to those derived from the coexisting planktonic population. Twenty-nine morphologically distinct bacterial strains were isolated from a 6-month-old biofilm, established in a glass tank under high-shear conditions, and 15 distinct strains were isolated from the associated re-circulating water. All 44 strains were identified to genus or species level by 16S rDNA sequencing. The 29 biofilm strains belonged to 14 genera and 23.4% of all the possible pair-wise combinations coaggregated. The 15 planktonic strains belonged to seven genera and only 5.8% of all the possible pair-wise combinations coaggregated. Therefore, compared to the planktonic population, a greater proportion of the biofilm strains coaggregated. It is proposed that coaggregation influences biofilm formation and species diversity in freshwater under high shear. PMID- 12644240 TI - Agrobacterium tumefaciens-mediated transformation as a tool for insertional mutagenesis in the symbiotic ectomycorrhizal fungus Hebeloma cylindrosporum. AB - We transformed haploid mycelium of Hebeloma cylindrosporum via Agrobacterium tumefaciens and optimised the procedure to develop a new tool for insertional mutagenesis in this fungus. Southern blot analysis of 83 randomly selected transformants showed that they all contained plasmid inserts. Each of them showed a unique hybridisation pattern, suggesting that integration was random in the fungal genome. Sixty percent of transformants obtained in the presence of bacteria pre-treated with acetosyringone integrated a single transferred DNA copy. Thermal asymmetric interlaced polymerase chain reaction allowed us to recover the left border and the right border junctions in 85% and 15% of transformants analysed, respectively. Results show that A. tumefaciens-mediated transformation may be a powerful tool for insertional mutagenesis in H. cylindrosporum. PMID- 12644241 TI - Purification of the 67-kDa lectin-like glycoprotein of Trypanosoma cruzi, LLGP 67, and its evaluation as a relevant antigen for the diagnosis of human infection. AB - In the present work we propose a simple method for affinity purification of the 67-kDa lectin-like glycoprotein (LLGP-67) from Trypanosoma cruzi, the causative agent of Chagas' disease. The LLGP-67, which presents galactose binding activity and participates in the host cell recognition process, was previously purified by methods based on its interaction with galactose residues on erythrocytic membranes. We describe herein results showing that this protein can be purified from T. cruzi in a direct way using non-derivatized agarose as a chromatographic ligand. We also demonstrate the relevance of LLGP-67 as an antigen for human diagnosis of chagasic infection. Sensitivity and specificity for this antigen were calculated, being 98 and 98.11% respectively. PMID- 12644242 TI - DNA microarray analysis of Bacillus subtilis sigma factors of extracytoplasmic function family. AB - Target gene candidates of the seven extracytoplasmic function (ECF) sigma factors of Bacillus subtilis have been surveyed using DNA microarray analysis of mRNA extracted from cells grown in Luria-Bertani broth, in which an ECF sigma factor gene was placed under the control of the spac promoter on multicopy plasmid pDG148 and overexpressed. The number of target candidates for each of the sigma factors varied greatly, and a total of 278 genes were selected. Interestingly, the above target gene candidates shared only one gene out of 94 target genes of the general stress sigma B that have been reported in the literature thus far. Furthermore, lacZ-fusion experiments based on the results of DNA microarray analysis indicated that each ECF sigma factor directs transcription of its own operon, with the exception of sigZ. The DNA microarray data collected in this study are available at the KEGG Expression Database web site (http://www.genome.ad.jp/kegg/expression/). PMID- 12644243 TI - Cytoarchitecture and fiber pattern of the superior colliculus are disrupted in the Shaking Rat Kawasaki. AB - Shaking Rat Kawasaki (SRK) is a Reelin-deficient rat, that shows significant cytoarchitectural abnormalities in the cerebral and cerebellar cortices in a similar manner to the reeler malformation. In the present study, we investigated the cytoarchitecture and myeloarchitecture of the superior colliculus (SC) of this mutant rat. The Nissl staining clearly showed that neuronal components in the superficial layers of the SC in SRK rat were intermingled with each other and that the boundaries between these superficial layers were blurred. The MBP immunohistochemistry showed an abnormal fiber pattern in the superficial layers of the SC of this mutant rat. In the normal rat, myelinated fibers passed rostrocaudally through the optic layer, and only a few myelinated fibers were recognized in the uppermost two layers, i.e., the zonal and superficial gray layers. By contrast, in SRK rat, the myelinated fibers were distributed throughout the entire thickness of the superficial layers of the SC. Anterograde labeling of retinotectal fibers with an injection of Cholera Toxin subunit B into the retina revealed that this abnormal fiber pattern was associated with the anomalous course of the retinotectal fibers. No distinct differences in the cytoarchitecture and fiber pattern in the deep layers of the SC were seen. In conclusion, the present study demonstrated that the cytoarchitecture and fiber patterning in the superficial layers of the SC were disrupted in SRK rat, suggesting that Reelin protein regulates the formation of the superficial layers of the SC. PMID- 12644244 TI - Differential expression of fibroblast growth factor receptors in the developing murine choroid plexus. AB - The choroid plexuses (CPs) are specialised secretory organs situated within the ventricles of the brain involved in the production of cerebrospinal fluid (CSF) and the maintenance of the blood-CSF barrier. Abnormal function of the CPs can lead to hydrocephalus and raised intracranial pressure, pathologies frequently observed in certain craniofacial syndromes caused by single point mutations in fibroblast growth factor receptors (FGFRs). At present, relatively little is known about the embryonic CPs in terms of gene or protein expression, function as the brain develops or on the potential role of FGFRs within this context. Given the limited information available on the regulation of FGFRs during development of the CPs and periventricular tissues, we have carried out a detailed analysis of the localisation of FGFR1, 2, 3 and 4 proteins in these regions of the murine embryo from the time of formation of the CP in the third ventricle at E12.5 throughout the second half of gestation, and examined the expression of different FGFR isoforms at E12.5 by RT-PCR. We show here that FGFR1 and FGFR4 are expressed in murine CPs at E12.5 but not at E15.5 or E18.5, suggesting a role for the signaling pathways transduced by these receptors at early stages of CP development. In contrast, FGFR2 expression is maintained throughout CP development, indicating that this receptor may play a role in the function of immature and mature CP. Also FGFR3 is detected at each developmental stage studied, but surprisingly its expression appears confined to the nuclei of CP cells, suggesting that FGFR3 in the CP does not respond to extracellular FGFs but may act in intracrine fashion. PMID- 12644245 TI - Effects of interleukin-10 on neonatal excitotoxic brain lesions in mice. AB - Interleukin-10 markedly reduces production of proinflammatory cytokines by activated microglia or macrophages and downregulates the expression of activating molecules on these cells. In studies performed in adults or in cell cultures, interleukin-10 protected against hypoxic-ischemic neuronal death and against lipopolysaccharide-mediated oligodendrocyte cell death. Furthermore, it was recently shown that interleukin-10 counteracts metabolic and microcirculatory effects of hypoxia-ischemia in the perinatal pig brain. Intracerebral injection of the glutamatergic analogue ibotenate to newborn mice induces cortical plate and white matter lesions mimicking the brain damage associated with cerebral palsy, and pretreatment with proinflammatory cytokines such as interleukin-1-beta or with interleukin-9 significantly exacerbates these lesions. The present study evaluated the influence of interleukin-10 on ibotenate-induced brain lesions in newborn mice under basal conditions or after exposure to cytokines. Intraperitoneal injection of interleukin-10 for 3 days following ibotenate significantly reduced the size of excitotoxic brain lesions. Intraperitoneal injection of neutralizing anti-interleukin-10 antibody for 3 days following ibotenate had no detectable effect and no difference in ibotenate-induced brain lesion size was found between wild type pups and pups deleted for the interleukin 10 gene, suggesting that endogenous interleukin-10 in newborn mice may have limited effects. Co-administration of intracerebral ibotenate and interleukin-10 had no detectable effect, arguing against a direct neuroprotective effect of interleukin-10 on neurons. While pretreatment with intraperitoneal interleukin-10 alone had no detectable effect on excitotoxic brain lesions, interleukin-10 given with interleukin-1-beta pretreatment blunted the toxic effects of interleukin-1 beta. On the other hand, combined pretreatment with IL-9 and anti-IL-10 antibody largely reversed the exacerbating effect of IL-9 on excitotoxic brain lesions. Altogether, these data suggest that, in newborn mice, exogenous interleukin-10 can be neuroprotective when acting in an inflammatory context. PMID- 12644246 TI - Glycogen synthase kinase (GSK)-3beta levels and activity in a neurodevelopmental rat model of schizophrenia. AB - We have previously reported reduced GSK-3beta protein levels and GSK-3 total (alpha + beta isoforms) activity in postmortem frontal cortex of schizophrenic patients. We now studied whether GSK-3beta is altered in the frontal cortex of rats with the neonatal excitotoxic hippocampal lesion used as a model of schizophrenia. Rats were infused with ibotenic acid (or artificial CSF in controls) bilaterally into the ventral hippocampus (VH) at postnatal day 7, then killed at postnatal day 35 (pre-puberty) or 56 (post-puberty). GSK-3beta protein levels were reduced in the frontal cortex of the lesioned rats as compared to sham animals; post-hoc comparisons revealed that the reduction was statistically significant at a pre-pubertal age. Total GSK-3 (alpha + beta) activity was not different between lesioned and sham rats at any age. These results demonstrate that reduced frontal cortical GSK-3beta levels may occur as a result of neonatal hippocampal damage and suggest that this animal model may be utilized to study the mechanism of GSK-3 reduction in schizophrenia, a disorder in which postmortem changes in GSK-3 were found. PMID- 12644247 TI - Cajal-Retzius cells in the mouse: transcription factors, neurotransmitters, and birthdays suggest a pallial origin. AB - Cajal-Retzius cells are reelin-secreting neurons found in the marginal zone of the mammalian cortex during development. Recently, it has been proposed that Cajal-Retzius cells may be generated both early and late in corticogenesis, and may migrate into the cortex from proliferative zones in the subpallium (lateral ganglionic eminence and medial ganglionic eminence) or cortical hem. In the present study, we used reelin as a marker to study the properties of Cajal Retzius cells, including their likely origins, neurotransmitters, and birthdates. In double labeling experiments, Cajal-Retzius cells (reelin(+)) expressed transcription factors characteristic of pallial neurons (Tbr1 and Emx2), contained high levels of glutamate, were usually calretinin(+), and were born early in corticogenesis, on embryonic days (E)10.5 and E11.5. Tbr1(+) cells in the marginal zone were almost always reelin(+). The first Cajal-Retzius cells (Tbr1(+)/reelin(+)) appeared in the preplate on E10.5. In contrast, interneurons expressed a subpallial transcription factor (Dlx), contained high levels of GABA, were frequently calbindin(+), and were born throughout corticogenesis (from E10.5 to E16.5). Interneurons (Dlx(+)) first appeared in the cortex on E12.5. Our results suggest that the marginal zone contains two main types of neurons: Cajal Retzius cells derived from the pallium, and migrating interneurons derived from the subpallium. PMID- 12644248 TI - A Golgi-Cox morphological analysis of neuronal changes induced by environmental enrichment. AB - Exposure to an enriched environment (EE), consisting of a combination of increased exercise, social interactions and learning, has been shown to produce many positive effects in the CNS. In this study, we use a Golgi-Cox analysis to examine and dissect the role of various components of the enriched environment on two measures of neuronal growth: total cell volume and total dendritic length in four regions of the brain. In the hippocampus, CA1 and dentate gyrus cells, animals raised in an enriched environment demonstrate significant morphological change. These changes were not observed in layer V pyramidal neurons of the cerebral cortex or spiny neurons located in the striatum. To determine if one or more of the individual components of the EE were responsible for the changes in neuronal morphology, we examined mice raised with free access to exercise wheels. In these mice, no morphological changes were observed. These results suggest that changes in the CA1 and dentate gyrus morphology were a result of alterations in the animal's environment and not an increase in motor activity. PMID- 12644249 TI - Visual deprivation effects on the s100beta positive astrocytic population in the developing rat visual cortex: a quantitative study. AB - After birth, exposure to visual inputs modulates cortical development, inducing numerous changes of all components of the visual cortex. Most of the cortical changes thus induced occur during what is called the critical period. Astrocytes play an important role in the development, maintenance and plasticity of the cortex, as well as in the structure and function of the vascular network. Dark reared Sprague-Dawley rats and age-matched controls sampled at 14, 21, 28, 35, 42, 49, 56 and 63 days postnatal (dpn) were studied in order to elucidate quantitative differences in the number of positive cells in the striate cortex. The astrocytic population was estimated by immunohistochemistry for S-100beta protein. The same quantification was also performed in a nonsensory area, the retrosplenial granular cortex. S-100beta positive cells had adult morphology in the visual cortex at 14 dpn and their numbers were not significantly different in light-exposed and nonexposed rats up to 35 dpn, and were even higher in dark reared rats at 21 dpn. However, significant quantitative changes were recorded after the beginning of the critical period. The main finding of the present study was the significantly lower astroglial density estimated in the visual cortex of dark-reared rats over 35 dpn as well as the lack of difference at previous ages. Our results also showed that there were no differences when comparing the measurements from a nonsensory area between both groups. This led us to postulate that the astrocytic population in the visual cortex is downregulated by the lack of visual experience. PMID- 12644251 TI - Caveolin-3 expression during early chicken development. AB - Caveolin-3, a protein that is correlated with caveolae, is found in muscle cells, especially during their differentiation. Although the distribution of caveolin-3 has been studied in cases such as adult and late embryonic mammalians, the expression of caveolin-3 has not been clearly defined during chicken development. In this study, we detected intense caveolin-3 immunoreactivity (IR) as early as embryonic day 4 (E4), most of the signals were localized within the neural tube and myotome. While IRs in the brain occurred in radial glia at E6, these intensities were reduced to an almost undetectable level at E8. In the case of muscle cells, the exclusive localization of caveolin-3 in the cytoplasmic membrane was detected even at E11, much earlier than in mammalian muscle tissues. Although the caveolin-3 IR pattern was similar to that reported by previous studies, we found some interesting mismatches in the case of avian tissues. Although we are unable to explain caveolin-3 expression patterns in the early embryonic stages, this study could provide a basis for further study on the function of caveolin-3 in avian embryogenesis. PMID- 12644250 TI - Methylmercury decreases NGF-induced TrkA autophosphorylation and neurite outgrowth in PC12 cells. AB - Neurotrophin signaling through Trk receptors is important for differentiation and survival in the developing nervous system. The present study examined the effects of CH(3)Hg on (125)I-nerve growth factor (NGF) binding to the TrkA receptor, NGF induced activation of the TrkA receptor, and neurite outgrowth in an in vitro model of differentiation using PC12 cells. Whole-cell binding assays using (125)I NGF revealed a single binding site with a K(d) of approximately 1 nM. Methylmercury (CH(3)Hg) at 30 nM (EC(50) for neurite outgrowth inhibition) did not affect NGF binding to TrkA. TrkA autophosphorylation was measured by immunoblotting with a phospho-specific antibody. TrkA autophosphorylation peaked between 2.5 and 5 min of exposure and then decreased but was still detectable at 60 min. Concurrent exposure to CH(3)Hg and NGF for 2.5 min resulted in a concentration-dependent decrease in TrkA autophosphorylation, which was significant at 100 nM CH(3)Hg. To determine whether the observed inhibition of TrkA was sufficient to alter cell differentiation, NGF-stimulated neurite outgrowth was examined in PC12 cells after exposure to 30 nM CH(3)Hg, a concentration that inhibited TrkA autophosphorylation by approximately 50%. For comparison, a separate group of PC12 cells were exposed to a concentration of the selective Trk inhibitor K252a (30 nM), which had been shown to produce significant inhibition of TrkA autophosphorylation. Twenty-four hour exposure to either CH(3)Hg or K252a reduced neurite outgrowth to a similar degree. Our results suggest that CH(3)Hg may inhibit differentiation of PC12 cells by interfering with NGF-stimulated TrkA autophosphorylation. PMID- 12644252 TI - Developing patterns of nitric oxide synthesizing neurons in the rat striatum: histochemical analysis. AB - The prenatal and postnatal development of NADPH-diaphorase (NADPH-d)/neuronal nitric oxide synthase (nNOS) positive neurons was studied in the striatum of rats. NADPH-d was demonstrated enzyme histochemically and nNOS immunohistochemically using a polyclonal antibody. NADPH-d neurons appeared in the ventrolateral part of the striatum on embryonic day 18 (E18). Thereafter, the number of NADPH-d neurons increased and began to distribute homogeneously in the striatum. The density of NADPH-d neurons became highest at postnatal day 5 (P5) and then decreased as the volume of the striatum continued to increase. The number of NADPH-d neurons reached its peak around 3-4 weeks after birth. The sizes of NADPH-d neurons were measured. The NADPH-d neurons grew larger until P14 (mean area 260 microm(2)) and became smaller thereafter (mean area 170 microm(2)). Patches of high NADPH-d activity and tyrosine hydroxylase (TH) immunoreactivity were also examined in the developing striatum. The distributions of NADPH-d patches overlapped with those of TH-immunoreactive patches by P10. The spatiotemporal appearance of nNOS and overlapping of nNOS patchy distribution with TH point to an important role of NO and to an interaction between nNOS and DA fibers during development of the striatum. PMID- 12644253 TI - Magnetic resonance proton spectroscopy and diffusion weighted imaging of chick embryo brain in ovo. AB - Metabolic compensatory mechanisms may partly account for the decreased vulnerability to hypoxia observed in the developing brain. We used proton magnetic resonance spectroscopy and diffusion-weighted imaging to measure the cerebral concentrations of lactate and other metabolites, as well as the apparent diffusion coefficient (ADC) of tissue water, before, during and after hypoxia in anaesthetised chick embryos in ovo. Reducing the inspired oxygen concentration to 8% for 40 min caused a significant rise in both mean (+/-S.D.) lactate:creatine and alanine:creatine ratios from 0.58 (0.41) to 1.56 (0.56) and 0.14 (0.14) to 0.29 (0.17), respectively. Under similar hypoxic conditions, ADC did not change from a mean baseline value of 0.95 (0.09), but did fall to 0.40 (0.12) x 10(9) mm(2) s(-1) with further stepwise reductions in oxygenation. Moderate hypoxia increases lactate concentration in the developing chick brain without compromising cellular energy metabolism. PMID- 12644254 TI - Chronic neonatal isolation stress enhances cocaine-induced increases in ventral striatal dopamine levels in rat pups. AB - Cocaine-induced increases in ventral striatal dopamine levels are enhanced in adult rats previously exposed to chronic stress. In neonatal rats, isolation from dam, nest, and siblings is stressful as evidenced by elevated corticosterone levels, an effect that increases with chronic isolation. Whether chronic neonatal isolation cross-sensitizes to cocaine leading to greater increases in ventral striatal dopamine levels was assessed in this study. Litters were assigned to isolate or non-handled conditions. After culling on postnatal Day 1 (PN1), pups in the neonatal isolation condition were isolated individually for 1 h/day on PN days 2-9 while pups in the non-handled condition were left undisturbed. On PN10, pups were implanted with probes aimed at the ventral striatum. Baseline measures of dopamine and its metabolite, DOPAC, were obtained. Separate groups of male and female pups were then administered 0, 2.5, 5.0, or 10 mg/kg cocaine and samples were collected for 2-h. Isolate pups showed greater cocaine-induced increases in ventral striatal dopamine levels than non-handled pups. However, DOPAC levels did not differ by isolation condition or gender. Neonatal isolation-induced increases in the effects of cocaine on ventral striatal dopamine levels are consistent with our previous study using amphetamine in 10-day-old pups and show that chronic stress sensitizes the dopamine response to psychostimulants in infant rats. PMID- 12644255 TI - Changes in cell cycle parameters and cell number in the rat midbrain during organogenesis. AB - We employed 5-bromo-2'-deoxyuridine (BrdU) labeling to identify in vivo changes in the cell cycle patterns of the rat midbrain during the major period of midbrain organogenesis, gestational days (gd) 11 to 16. We also used quantitative stereology to determine changes in absolute cell numbers during these gestational time points. Between gd 12 and 16, the length of S-phase did not change significantly while the fraction of cycling cells decreased from 73 to 11%. The average cell cycle length was determined to be 15 h on gd 12 and 17 h on gd 16, the difference not being statistically significant. The cell number in the midbrain increased from 1.3E5 cells on gd 11 to 1.7E7 cells on gd 16. On gd 12 and gd 13, there was a significant negative correlation between litter position and midbrain cell number, the effect diminishing on later days of gestation. The combined use of quantitative stereology and flow cytometry to study brain development represents a novel application that allows for simultaneous evaluation of changes in cell proliferation kinetics and the resulting effect of those kinetic changes on embryonic midbrain development. PMID- 12644256 TI - Gene expression of EXT1 and EXT2 during mouse brain development. AB - Heparan sulfate (HS) and heparan sulfate proteoglycans (HSPGs) play significant roles in various biological processes. There is a wealth of circumstantial and experimental evidence suggesting the roles of HS in mammalian neural development. HS synthesis is governed by a series of enzymes. Among them, two enzymes, EXT1 and EXT2, catalyze polymerization of glucuronic acid and N-acetylglucosamine, the crucial step of HS synthesis. To obtain insight into the roles of HS in neural development, we examined the spatiotemporal expression patterns of EXT1 and EXT2 during mice brain development. RT-PCR analyses showed that expression of EXT1 and EXT2 peaks during early postnatal period in the cerebrum and around birth in the cerebellum. In situ hybridization revealed that in the embryonic brain, EXT1 and EXT2 were localized primarily in the neuroepithelial cells surrounding the lateral ventricles, the mesencephalic vesicle, and the fourth ventricle. In the early postnatal stage, intense expression of EXT1 and EXT2 was observed in the cerebral cortex and the hippocampus formation. In the postnatal cerebellum, expression of EXT1 and EXT2 was mainly observed in external and internal granular layers. Our results demonstrate that EXT1 and EXT2 are highly expressed in the developing brain, and that their expression is developmentally regulated, suggesting that HS is involved in various neurodevelopmental processes. PMID- 12644257 TI - Glucose-lowering effect of beta-phenylpyruvate in neonatal mice: a possible mechanism for phenylketonuria-related neurodegenerative changes. AB - Following beta-phenylpyruvate injection, mice developed hypoglycemia clinically manifested as tachypnea, tremor, convulsions and death. To further investigate, neonatal mice were injected with beta-phenylpyruvate and their blood glucose determined and brain histology assessed. beta-Phenylpyruvate-injected mice exhibited higher mortality and neurophysiological changes as compared with controls, although without evidence of neural cell death. Accordingly, we hypothesize that the central neural damage in phenylketonuria might be caused by these recurrent beta-phenylpyruvate-induced hypoglycemic events. PMID- 12644258 TI - Localization of cartilage linking protein 1 during primary neurulation in the chick embryo. AB - Primary neurulation is a form-shaping event during the early development of the vertebrate embryo in which the neural plate is rolled up into the neural tube, the rudiment of the central nervous system. In an effort to identify genes specifically expressed in tissues lateral to the chick neural plate--tissues known to generate extrinsic forces for primary neurulation--we designed a subtractive scheme and identified a positive clone as the gene encoding chick cartilage linking protein 1 (CRTL1). CRTL1 (also known as link protein) is a small glycoprotein of the extracellular matrix that was originally identified for its role in stabilizing aggregates of aggrecan and hyaluronan in cartilage. In addition to being expressed in cartilage, CRTL1 is also immunolocalized in several noncartilaginous tissues as assessed with the 4B6 monoclonal antibody. Using the 4B6 antibody and the G9 riboprobe derived from our subtraction, we report the detailed distribution of CRTL1 protein and crtl1 transcripts during primary neurulation in chick embryos. This report emphasizes and briefly discusses important differences between the RNA expression pattern and the domains of accumulation of the protein. CRTL1 prominently accumulates in the basal lamina of the epidermal ectoderm just lateral to the neural plate. Based on the crucial role of the interface between this tissue and the neuroepithelium in the formation of the neural folds, and because of the biophysical role of hyaluronan in tissue morphogenesis, we propose that crtl1 represents is an excellent candidate neurulation gene, worthy of further study. PMID- 12644259 TI - The associative striatum: cortical and thalamic projections to the dorsocentral striatum in rats. AB - Corticostriatal projections to the dorsocentral striatum (DCS) were investigated using retrograde fluorescent axonal tracing. The DCS is of interest because of its role in directed attention and recovery from multimodal hemispatial neglect following cortical lesions of medial agranular cortex (AGm), an association area that is its major source of cortical input. A key finding was that the multimodal posterior parietal cortex (PPC) also contributes substantial input to DCS. This is significant because PPC and AGm are linked by corticocortical connections and are both critical components of the circuitry involved in spatial processing and directed attention. Other cortical areas providing input to DCS include visual association areas, lateral agranular cortex and orbital cortex. These areas also have reciprocal connections with AGm and PPC. Less consistent labeling was seen in somatic sensorimotor areas FL, HL and Par 1. Thalamic afferents to DCS are prominent from the intralaminar, ventrolateral, mediodorsal, ventromedial, laterodorsal (LD) and lateral posterior (LP) nuclei. Collectively, these nuclei constitute the sources of thalamic input to cortical areas AGm and PPC. Nuclei LD and LP are only labeled with injections in dorsal DCS, the site of major input from PPC, and PPC receives its thalamic input from LD and LP. We conclude that DCS receives inputs from cortical and thalamic areas that are themselves linked by corticocortical and thalamocortical connections. These findings support the hypothesis that DCS is a key component of an associative network of cortical, striatal and thalamic regions involved in multimodal processing and directed attention. PMID- 12644260 TI - ATP modulation of sodium currents in rat dorsal root ganglion neurons. AB - The modulation of tetrodotoxin-sensitive (TTX-S) and slow tetrodotoxin-resistant (TTX-R) sodium currents in rat dorsal root ganglion neurons by ATP was studied using the whole-cell patch-clamp method. The effects of ATP on two types of sodium currents were either stimulatory or inhibitory depending on the kinetic parameters tested. At a holding potential of -80 mV ATP suppressed TTX-S sodium currents when the depolarizing potential was positive to -30 mV but it increased them when the depolarizing potential was negative to -30 mV. At the same holding potential slow TTX-R sodium currents were always increased by ATP regardless of the depolarizing potential. In both types of sodium currents ATP shifted both the conductance-voltage relationship curve and the steady-state inactivation curve in the hyperpolarizing direction, and accelerated the time-dependent inactivation. ATP decreased the maximum conductance of TTX-S sodium currents but increased that of slow TTX-R sodium currents. The results suggest that ATP would decrease the excitability of neurons with TTX-S sodium channels but would increase that of neurons with slow TTX-R sodium channels. The effects of ATP on sodium currents were preserved in the presence of a G-protein inhibitor, GDP-beta-S, or purinergic antagonists, suramin and Reactive Blue-2, suggesting that purinergic receptors might not be involved in ATP modulation of sodium currents. PMID- 12644261 TI - Sensory properties of articular afferents following Mycoplasma arthritis in the chicken. AB - The physiological properties of joint capsule mechanical nociceptors of monoarthritic chickens (Gallus domesticus) were studied by recording the electrical activity from single C (Group IV) and A-delta (Group III) fibres dissected from the parafibular nerve. By injecting live Mycoplasma gallisepticum cultures into the ankle joint a typical mycoplasma arthritis was induced which was restricted to a single joint. During the early stage of the disease (7-21 days after infection) there was histopathological evidence of an acute synovitis and the fibres showed evidence of sensitisation. Sensitisation was observed in the significantly increased receptive field size, decreased response thresholds, increased response to joint movement both noxious and innocuous, but only in the C-fibres was there an increase in spontaneous activity. During the more chronic stage of the disease (49-56 days after infection) there was pathological evidence of prolonged synovitis but the sensory fibres responded normally to mechanical stimulation and joint movement. These changes in sensitivity of the joint capsule mechanical nociceptors provides peripheral neural evidence of possible pain experienced during the acute stage of the disease but not at the chronic stage when the disease might be in period of remission. The absence of any clear correlation between pathology and receptor activity demonstrates the difficulty of trying to predict nociceptive consequences in animals on the basis of histopathology. PMID- 12644262 TI - A study on the mechanism by which sodium nitroprusside, a nitric oxide donor, applied to the anteroventral third ventricular region provokes facilitation of vasopressin secretion in conscious rats. AB - We reported previously that sodium nitroprusside (SNP) applied to the anteroventral third ventricular region (AV3V), a pivotal area for autonomic functions, facilitates vasopressin (AVP) secretion in conscious rats. The aim of this study was to pursue the problems of whether nitric oxide (NO) generated from the agent may be responsible for the phenomenon, and whether it may be mediated by cyclic guanosine monophosphate (cGMP), the biosynthesis of which could reportedly be activated by NO. The infusion of SNP into the AV3V of conscious rats produced dose-related increases in plasma AVP, the maximal responses of which appeared at 5 min. Blood pressure and heart rate tended to rise at 15 min. The plasma osmolality, sodium, potassium or chloride did not show marked alteration following the SNP administration. Although the SNP solution was hypertonic and hypernatremic, AV3V application of hypertonic saline with a relatively higher osmolality and an equal sodium level was significantly less effective in augmenting plasma AVP. When injected into the lateral ventricle, SNP did not change plasma AVP and reduced arterial pressure, different from the results provoked by the AV3V application. The rise in plasma AVP in response to the AV3V application of SNP was diminished by preadministration of hemoglobin, a scavenger of NO, that did not affect the responses of the other variables. In contrast, pretreatment with methylene blue, an agent capable of antagonizing the potency of NO to activate guanylate cyclase, did not attenuate but potentiated the responses of both plasma AVP and arterial pressure to the AV3V infusion of SNP. Hemoglobin or methylene blue given alone into the AV3V did not affect any of the variables monitored. On the other hand, the AV3V injection of 8-bromo cGMP, a stable analogue of cGMP, was not potent for causing a significant rise in plasma AVP, in contrast to the notable AVP-enhancing effect of 8-bromo cAMP. Arterial pressure and heart rate were elevated by both of these agents, whereas the remaining variables were not altered. Histological inspection indicated that the infusion sites of the drugs in the AV3V had included areas such as the organum vasculosum of the lamina terminalis, median preoptic nucleus, medial preoptic nucleus and periventricular nucleus. On the basis of these results, we concluded that the AVP secretion prompted by the AV3V application of SNP may be attributable to NO, whereas its well-known ability to stimulate guanylate cyclase activity may hardly contribute to this phenomenon. PMID- 12644263 TI - Sciatic nerve axotomy in aged rats: response of motoneurons and the effect of GM1 ganglioside treatment. AB - The number, size, and staining intensity of choline acetyltransferase (ChAT) immunopositive cells in the retrodorsal lateral nucleus (RDLN) of the spinal cord were studied in young (3-5 months old) and aged (22-24 months old) rats following left sciatic nerve distal transection (axotomy) and treatment with GM1 ganglioside. The cell size and the ChAT immunostaining density were decreased in the RDLN of non-manipulated as well as in the contralateral intact side of axotomized aged rats. Axotomy had no effect on the number of RDLN motoneurons in both aged and young rats. In the young rats, there was a decrease in the size of motoneurons 7 days post-axotomy and a partial spontaneous recovery occurred by 21 days. Axotomy did not reduce further the size of aged motoneurons, however. The ChAT staining intensity of the axotomized RDLN declined in both age groups after 7 days, and there was spontaneous near normal recovery by 21 days. In the aged rats, GM1 administration for 7 days corrected the cell size and ChAT immunoreactivity of the contralateral intact RDLN. With regard to axotomized RDLN neurons, 7 days of GM1 restored the cell size but not the ChAT immunostaining in young animals. The same treatment schedule, however, corrected both cell size and staining in aged rats. Administration of GM1 for 21 days had no further effect on the morphometric parameters of the axotomized motoneurons in aged rats, but slightly enhanced the recovery of ChAT immunostaining in young rats. Thus, it appears that GM1 facilitates the phenotypic recovery of RDLN motoneurons during aging and after axotomy. PMID- 12644264 TI - Quantitative comparisons of corticothalamic topography within the ventrobasal complex and the posterior nucleus of the rodent thalamus. AB - To compare the topographic precision of corticothalamic projections to the ventrobasal (VB) complex and the medial part of the posterior (POm) complex, different anterograde tracers were placed in neighboring parts of the primary (SI) and secondary (SII) somatosensory cortical areas. The location of labeled corticothalamic terminals and their beaded varicosities were plotted, and the digital reconstructions were analyzed quantitatively to determine the extent of overlapping projections from the cortical injection sites. Among animals that received all tracer injections in SI cortex, tracer overlap in the thalamus varied according to the proximity of the cortical injection sites. Regardless of which combination of somatic representations were injected in SI, within each animal the amount of tracer overlap in POm was similar to that observed in VB, and a matched-sample statistical analysis failed to reveal significant differences in the proportion of the labeled regions that contained overlapping projections from the injected cortical sites. Among those animals in which the tracers were injected into the whisker representations of SI and SII, the amount of tracer overlap in the thalamus was not affected by the proximity of the cortical injection sites. Instead, tracer overlap appeared to be related to the degree of somatotopic correspondence. Furthermore, within each of these animals, the amount of tracer overlap in POm was similar to that found in the VB complex. These results indicate that POm has a well-defined topographic organization that is comparable to the degree of topography observed in the VB complex. PMID- 12644265 TI - Molecular cloning and functional characterization of the OCTN2 transporter at the RBE4 cells, an in vitro model of the blood-brain barrier. AB - The transport of L-carnitine (4-N-trimethylamino-3-hydroxybutyric acid), a compound known to be transported by the organic cation transporter/carnitine transporter OCTN2, was studied in immortalized rat brain endothelial cells (RBE4). The cells were found to take up L-carnitine by a sodium-dependent process. This uptake process was saturable with an apparent Michaelis-Menten constant for L-carnitine of 54+/-10 microM and a maximal velocity of 215+/-35 pmol/mg protein/h. Besides L-carnitine, the cells also took up acetyl-L-carnitine and propionyl-L-carnitine in a sodium-dependent manner and TEA in a sodium independent manner. RT-PCR with primers specific for the rat OCTN2 transporter revealed the existence of OCTN2 mRNA in RBE4 cells. Screening of a cDNA library from RBE4 cells with rat OCTN2 cDNA as a probe identified a positive clone which showed, when expressed in HeLa cells, the functional characteristics of OCTN2. The HeLa cells expressing the RBE4 OCTN2 cDNA showed a sixfold increase in L carnitine uptake and a fourfold increase in TEA uptake in a sodium-containing buffer. Typical inhibitors for organic cation transporters (e.g. MPP(+) or TEA) showed an inhibitory effect on the transport of L-carnitine and TEA into the transfected cells. Similarly, unlabeled L-carnitine inhibited the transport of [3H]-L-carnitine and [14C]TEA in transfected HeLa cells. It is concluded that RBE4 cells, a widely used in vitro model of the blood-brain barrier (BBB), express the organic cation/carnitine transporter OCTN2. PMID- 12644266 TI - Central distribution of neuronal cell bodies innervating the levator veli palatini muscle and associated pattern of myosin heavy chain isoform expression in rat. AB - The levator veli palatini (LVP) is a muscle that plays a very important role in the complex functions regulating velopharyngeal function. Although previous studies have indicated that the contraction properties of the LVP closely resemble those of the intrinsic laryngeal muscle, histological evidence has not yet been obtained. The LVP is generally considered to be innervated by the glossopharyngeal nerve, which contains efferent and afferent components. LVP motoneurons are localized in the nucleus ambiguus (Amb), and afferent neurons project into the bilateral regions of the nucleus of the solitary tract (NST). However, the position of neuronal cell bodies on afferent neurons has remained unknown. The present study examined serial muscle cross-sections using monoclonal antibodies specific for myosin heavy chain (MyHC), to characterize muscle fibers of the LVP, clarify the central distribution of LVP motoneurons within the Amb and afferent terminals within the NST, and elucidate the location of LVP afferent neuronal cell bodies. Clear separation was observed within the LVP between fibers containing only fast MyHC and others positive for both slow and fast MyHC. Horseradish peroxidase (HRP)-labeled motoneurons in the Amb were separated into rostral and caudal divisions, corresponding to the Botzinger complex and the rostral ventral respiratory group, respectively. HRP-labeled afferent neuronal cell bodies were observed in a glossopharyngo-vagal complex ganglion, and HRP labeled afferent terminals were observed in bilateral lateral regions of the NST. These results suggest a relationship between MyHC isoform expression and the central distribution of LVP motoneurons or central projections of afferent neurons, with regard to activity of the LVP during both inspiration and expiration. PMID- 12644267 TI - Developmental 3,4-methylenedioxymethamphetamine (MDMA) impairs sequential and spatial but not cued learning independent of growth, litter effects or injection stress. AB - Previously, we have shown that rats administered MDMA from postnatal (P) days 11 20 had reductions in body weight during the period of treatment and as adults they had deficits in sequential and spatial learning and memory. In the present study, to control for weight reductions, we used litters with double the number of offspring to induce growth restriction comparable to that of standard size litters treated with MDMA. Litters were treated twice daily from P11 to 20 with vehicle or MDMA (20 mg/kg) or only weighed. Males, but not females, exposed to MDMA had longer latencies and more errors in the Cincinnati water maze compared to males of the other treatments. In the Morris water maze (210 cm pool, 10x10 cm platform), the MDMA animals were impaired relative to all other treatments during acquisition. Only the MDMA females showed deficits when the platform was shifted to a new location, however, both MDMA males and females were impaired when the location of the platform was again shifted and a reduced platform (5x5 cm) used. No differences were observed in the ability to swim a straight channel, locate a platform with a cue, or the endocrine response to forced swim among the treatment groups. No differences were seen between animals injected with saline and those only weighed. The data suggest that factors, such as growth retardation, multiple injections, or the composition of the litter, do not affect the development of learning and memory impairments resulting from P11 to 20 MDMA exposure. The large litter approach offers a novel method to control for undernutrition during the preweaning period in rodents. PMID- 12644268 TI - Modulation of D1-like dopamine receptor function by aldehydic products of lipid peroxidation. AB - Growing evidence indicates that aldehydic products of lipid peroxidation play an important role in the pathophysiology of neurodegenerative disorders such as Parkinson's disease. In the present study, modulation of D1-like receptor binding and function by saturated alkanals and unsaturated alkenals, 4-hydroxynonenal (4 HNE) and trans-2-nonenal (nonenal), was examined in rat striatal membranes. The 4 HNE and nonenal were most effective in modulating both the specific D1-like receptor binding and function as measured by adenylate cyclase activation. Inactivation of receptor binding and the depression of adenylate cyclase activity were partially prevented by protection of the D1/D5-receptor with the agonist (R) SKF 38393 or the specific antagonist SCH 23390. 4-HNE inhibited adenylate cyclase activation by Gpp (NH)p and forskolin, indicating the modulation of Gsalpha and the catalytic subunit of adenylate cyclase, respectively. Our data suggests that aldehydic products of lipid peroxidation can directly modulate the binding and functional properties of D1/D5 receptors, as well as effector proteins within their signaling pathway. PMID- 12644269 TI - The effect of bone marrow stromal cells on neuronal differentiation of mesencephalic neural stem cells in Sprague-Dawley rats. AB - There are numerous parallels between the heamatolymphopoietic and nervous systems in terms of the mechanisms regulating their development. We proposed that neural stem cells (NSCs) may respond to the microenvironmental signals provided by bone marrow stromal cells (BMSCs) which regulate the differentiation and maturation of hematolymphopoietic stem cells. First, we isolated and proliferated BMSCs from the femur and tibia, and NSCs from the midbrain of Sprague-Dawley (SD) rats, and then investigated the effects of BMSCs on the differentiation of NSCs into neurons, astrocytes and oligodendrocytes by directly plating neurospheres on BMSC monolayers in serum-free conditions. The results confirmed that BMSCs induced NSCs to differentiate selectively into neurons. The percentage of neurons significantly increased in 7 days in vitro co-cultures of NSCs and BMSCs as compared to NSCs cultures alone. When the duration of the cultures was extended to 12 days in vitro, BMSCs enhanced the survival of neurons derived from these NSCs; our investigation then focused on the underlying mechanism for this effect of BMSCs. NSCs were cultured with BMSC conditioned-medium and co-cultured with membrane fragments of live BMSCs or paraformaldehyde fixed BMSCs, the inducing activity of BMSCs was solely detectable in BMSC conditioned-medium, indicating that soluble factors secreted by BMSCs were responsible for its effect on the neuronal differentiation of NSCs. Therefore, BMSCs may provide a powerful tool for therapeutic neurological applications. PMID- 12644270 TI - Central alpha-adrenergic receptors and corticotropin releasing factor mediate hemodynamic responses to acute cold stress. AB - Behavioral stress is likely to contribute to the development of hypertension in susceptible individuals. We reported that hemodynamic response patterns to acute startle vary and that those patterns predict the predisposition of rats to sustained stress-induced elevations in arterial pressure. Since considerable evidence suggests that central catecholamines and corticotropin releasing factor (CRF) contribute to the regulation of arterial pressure and the development of hypertension, we investigated the role of central alpha-adrenergic receptors and CRF in mediating different hemodynamic response patterns to acute cold water stress in conscious rats. Rats were instrumented for arterial pressure, heart rate and cardiac output determination and for intracerebroventricular (icv) administration of selective antagonists. After acclimation to a water tight cage, ice water (1 cm deep) was rapidly added then drained 1 min later. Although the early startle response to cold water stress elicited a pressor response in all rats, the hemodynamic response pattern varied between rats. Vascular responders (n=19) had an initial considerable increase in systemic vascular resistance and a decrease in cardiac output. In contrast, mixed responders (n=11) had a smaller increase in vascular resistance and an increase in cardiac output. Pretreatment with phentolamine (30 microgram/5 microliter, icv, n=8), prazosin (10 microgram/5 microliter, icv, n=12) or alpha-helical CRF(9-41) (10 microgram/5 microliter, icv, n=9) prevented the decrease in cardiac output elicited by acute cold water stress in vascular responders without affecting mixed responders. Yohimbine (3 microgram/5 microliter, icv, n=8) pretreatment did not alter hemodynamic responses. Therefore, we conclude that central alpha(1)-adrenoceptors and CRF mediate the specific hemodynamic response patterns to acute startle and may be responsible for the predisposition to develop hypertension in vascular responders. PMID- 12644271 TI - Influence of paradoxical sleep deprivation and cocaine on development of spontaneous penile reflexes in rats of different ages. AB - Recent studies have established that paradoxical sleep deprivation (PSD) and cocaine administration induce genital reflexes (penile erection and ejaculation) in adult and old rats. To determine whether the same effects would induce spontaneous genital reflexes in rats of different ages (30-90 days old), we administered with cocaine (7 mg/kg) or saline to rats after a 4-day period of PSD, or at the equivalent time-point in control animals, and penile erection and ejaculation were then evaluated. In PSD rats administered cocaine, erection was observed from 30 days old to 90 days old, when both genital reflexes reached a peak. Animals submitted to PSD and saline injection showed erection from 60 to 90 days old. None of the control (saline and cocaine) groups of any age displayed these behaviors. The effects of PSD on steroid hormone levels showed that, although testosterone levels increased with age, PSD caused a marked decrease in testosterone at all ages evaluated. Progesterone and corticosterone levels were higher in PSD groups than in the respective control groups. These findings suggest that the interaction of PSD and cocaine probably enhances dopaminergic transmission in the brain and may accelerate the development of genital reflexes in male rats. PMID- 12644273 TI - Signalling pathways involved in the short-term potentiation of dopamine release by BDNF. AB - Brain-derived neurotrophic factor (BDNF) has been shown to modulate synaptic plasticity in the corpus striatum in vitro by activation of the tyrosine kinase linked receptor, TrkB. However, the signalling pathways that mediate this modulation of plasticity are poorly understood. Three proteins mediating signalling pathways are activated by the binding of BDNF to TrkB: phosphoinositol 3 kinase (PI3K); Ras-MEK and phospholipase C-gamma (PLCgamma). The present study investigates which of these pathways are necessary for BDNF-mediated potentiation of synaptic output of dopamine from slices and synaptosomes of rat corpus striatum. The results indicate that activation of the PI3K and Ras-MEK pathways, but not PLCgamma, are involved. Inhibitors of transcription and translation had no effect on the potentiation of depolarisation-stimulated (15 mM KCl) dopamine release mediated by BDNF. PMID- 12644272 TI - Localization of AT(2) receptors in the nucleus of the solitary tract of spontaneously hypertensive and Wistar Kyoto rats using [125I] CGP42112: upregulation of a non-angiotensin II binding site following unilateral nodose ganglionectomy. AB - We have examined the binding distribution of a selective AT(2) receptor ligand [125I] CGP42112 in the brain of adult Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR). AT(2) receptor localization was also examined in the rat brainstem following unilateral nodose ganglionectomy. Specific [125I] CGP42112 binding was observed in discrete brain regions from both rat strains, including the nucleus of the solitary tract (NTS), and did not differ between WKY and SHR. [125I] CGP42112 binding in the NTS revealed an AT(2) receptor component that was displaceable by PD 123319 and Ang II (50-58%), as well as a non-angiotensin II receptor component (42-49%). Following unilateral nodose ganglionectomy, [125I] CGP42112 binding density on the denervated side of the NTS was increased approximately two-fold in both WKY and SHR. This increased [125I] CGP42112 binding density in the ipsilateral NTS was comprised of a greater non-angiotensin II component than that observed in the sham groups, since only approximately 30% was displaced by PD123319 and angiotensin II. Furthermore, [125I] CGP42112 also revealed high binding density on the denervated side in the dorsal motor nucleus and the nucleus ambiguus in both WKY and SHR. AT(2) receptor immunoreactivity was also visualised in the NTS of sham operated rats, but was not observed in the dorsal motor nucleus or the nucleus ambiguus, nor was it up-regulated following nodose ganglionectomy. These results demonstrate, for the first time, an AT(2) receptor binding site in the NTS, as well as a non-angiotensin II [125I] CGP42112 binding site. These studies also demonstrate that nodose ganglionectomy represents a useful model in which to study a non-angiotensin II [125I] CGP42112 binding site that is up-regulated following degeneration of afferent vagal nerves. PMID- 12644274 TI - Chemical kindling induced by pentylenetetrazol in histamine H(1) receptor gene knockout mice (H(1)KO), histidine decarboxylase-deficient mice (HDC(-/-)) and mast cell-deficient W/W(v) mice. AB - The role of brain histamine on seizure development of pentylenetetrazol (PTZ) induced kindling was examined in H(1)-receptor gene knockout (H(1)KO), histidine decarboxylase-deficient (HDC(-/-)) and mast cell-deficient (W/W(v)) mice. All H(1)KO, HDC(-/-) and W/W(v) mice had accelerated seizure development of PTZ induced kindling when compared to their respective wild-type mice. The daily PTZ kindling increased histamine content in the cortex and diencephalon of H(1)KO mice, whereas the histamine content in the diencephalon of W/W(v) mice was decreased. The present study indicates that histamine plays a suppressive role in seizure development through H(1)-receptors. PMID- 12644275 TI - Antisense knockdown of neuronal nitric oxide synthase antagonizes nitrous oxide induced behavior. AB - The behavioral effects of nitrous oxide (N(2)O) were antagonized by non-specific inhibitors of nitric oxide synthase (NOS). To identify the isoform of NOS involved in this response, mice were pretreated with an antisense oligodeoxynucleotide (AS-ODN) against neuronal NOS, and then tested in a light/dark exploration paradigm. The AS-ODN but not the mismatch ODN significantly antagonized N(2)O induced behavior and also reduced NOS activity in the cerebellum and hippocampus. These results implicate neuronal NOS in the N(2)O response. PMID- 12644276 TI - Glutamate receptor function in learning and memory. AB - The contribution of glutamate to synaptic transmission, plasticity and development is well established; current evidence is based on diverse approaches to decipher function and malfunction of this principal transmitter. With respect to learning and memory, we are now able to identify more specifically the role played by the three main glutamate receptor classes in learning and memory: centre stage is clearly the NMDA receptor, with overwhelming evidence proving its involvement in the actual learning process (encoding), throughout the animal kingdom. This is discussed with respect to many different types of learning. Evidence for the contribution of the AMPA receptors (AMPARs) is less clear-cut due to the general problem of specificity: block of AMPARs will shutdown neuronal communication, and this will affect various components essential for learning. Therefore, the role of AMPARs cannot be established in isolation. Problems of interpretation are outlined and a specific involvement of AMPARs in the regulation of neuronal excitation related to learning is proposed. Metabotropic glutamate receptors (mGluRs) may contribute very little to the actual acquisition of new information. However, memory formation appears to require mGluRs, through the modulation of consolidation and/or recall. Overall, mGluR functions seem variable and dependent on brain structure and learning task. PMID- 12644277 TI - Drinking attenuates the noradrenaline release in the lateral hypothalamic area induced by angiotensin II activation of the subfornical organ in rats. AB - Experiments were conducted to investigate the role of noradrenergic systems in the lateral hypothalamus area (LHA) in the water intake caused by injection of angiotensin II (ANG II) into the subfornical organ (SFO) in rats. Intracerebral microdialysis techniques were utilized to quantify the extracellular content of noradrenaline (NA) in the LHA. Microinjection of ANG II into the SFO significantly increased NA release in the LHA when water was not available for drinking. The increase in the release of NA in the LHA was significantly attenuated by water intake. In urethane-anesthetized rats, injections of ANG II into the SFO significantly enhanced the release of NA in the LHA that accompanied an elevation in mean arterial pressure (MAP). Intravenous administration of the alpha-agonist metaraminol, on the other hand, significantly decreased the NA release in the LHA that accompanied an increase in MAP, suggesting that the enhanced NA release in the LHA caused by ANG II into the SFO may be not mediated by increasing in arterial pressure. These results show the involvement of the noradrenergic systems in the LHA in the dipsogenic response induced by angiotensinergic activation of the SFO. PMID- 12644278 TI - Facilitation of male-like coital behavior in female rats by kindling. AB - Kindling is a model of epilepsy and brain plasticity. When applied to the medial preoptic area (MPOA) of non-copulating male rats kindling induces masculine sexual behavior. In order to test if kindling could facilitate male-like coital behavior in female rats, sexually naive females were ovariectomized and kindled in the amygdala (AMG) or the MPOA until an intermediate stage (between 1 and 3, MPOA1-3) or until stage 5 (MPOA5 group). Once kindling was established, females were treated with 2.5 mg/Kg of testosterone propionate (TP) for 15 days. Male like coital behavior was evaluated on days 5, 10 and 15 of treatment. Subjects were then injected with a TP dose of 5 mg/kg for 15 days and tested in the same way as with the lower dose. The number of mounts was significantly increased and the mount latency was significantly reduced in the MPOA1-3 group when tested 5 days after treatment with the low dose of TP. The same facilitation was observed in MPOA1-3 and MPOA5 groups on day 10 of treatment with the low dose of TP. When the animals were under the high dose of TP treatment, the number of intromissions was increased in all experimental groups (including the AMG kindled group) in comparison to sham animals. In a second experiment we evaluated if the facilitation of male-like coital behavior induced by kindling was produced by a modification of the response of the vomeronasal system to sexually relevant cues. Ovariectomized females were stimulated until they reached kindling stage 2, then they were treated with 2.5 mg/kg of TP for 5 days. After animals were exposed for 90 min to clean sawdust or sawdust soiled by estrous females they were perfused. Fos was detected by immunocytochemistry along the vomeronasal pathway. No differences were found in Fos responses between sham and MPOA kindled females. The facilitation of masculine sexual behavior observed in AMG kindled females may be a consequence of the propagation of the AD to other brain regions involved in the expression of masculine sexual behavior. We propose that masculine sexual behavior is facilitated in MPOA kindled female rats by local neural changes produced by this kind of stimulation without modifying the response of the vomeronasal system to sexually relevant cues. PMID- 12644279 TI - Light induces cortical activation and yawning in rats. AB - We examined the effects of light stimulation on cortical activation and yawning response in anesthetized, spontaneously breathing rats. Cortical activation was assessed by means of an electrocorticogram (ECoG) and yawning response was evaluated by monitoring an intercostal electromyogram as an index of inspiratory activity and a digastric electromyogram as an indicator of mouth opening. Light stimulation elicited an arousal shift in the ECoG to faster rhythms. This arousal response was followed by a single large inspiration with mouth opening, i.e. a yawning response. Higher light intensity significantly reduced the onset latency of the arousal/yawning response. Pretreatment with pyrilamine, an H1-histamine receptor antagonist, injected into the lateral ventricle blocked both the cortical activation and the yawning response induced by light stimulation, suggesting a role of brain histaminergic neurotransmission in modulating the light-induced arousal/yawning responses. PMID- 12644280 TI - Persistent behavioral and autonomic supersensitivity to stress following prenatal stress exposure in rats. AB - Prenatal restraint stress (PS) has been suggested as an attractive chronic model of anxiety. Here, we characterized the behavioral and autonomic responsivity to acute stress exposure in adult PS subjects. In Wistar rats, locomotor activity, as well as spontaneous behavior in an established animal model of anxiety, the elevated plus-maze, was unaffected by PS. However, the anxiogenic-like response normally seen on the plus-maze following a restraint stress was markedly potentiated in adult PS subjects, despite indistinguishable corticosterone responses. In addition, we assessed the tail skin temperature response to a mild social stressor, transient social mixing. The diazepam-sensitive, late phase of the temperature response was markedly potentiated in adult PS subjects. In summary, PS induces a persistent phenotype of increased behavioral and autonomic sensitivity to stress. This paradigm might serve as an attractive screening model for anti-anxiety compounds. PMID- 12644281 TI - Increases in avoidance responding produced by REM sleep deprivation or serotonin depletion are reversed by administration of 5-hydroxytryptophan. AB - Our objective was to directly compare the effects of rapid eye movement (REM) sleep deprivation (REMSD) and serotonin 5-hydroxytryptamine (5-HT) depletion on free-operant avoidance behavior in rats. These experiments were designed to determine if declining 5-HT levels observed during REMSD might mediate the increases in avoidance responding observed in REM sleep deprived rats. Rats were trained on a free-operant avoidance task. Following training, the animals were assigned to one of three sleep conditions (REMSD, tank control, or cage control). Animals in each sleep condition were exposed to four 5-HT manipulations: (a) saline plus saline; (b) p-chlorophenylalanine (PCPA) plus saline; (c) saline plus 5-hydroxytryptophan (5-HTP) and (d) PCPA plus 5-HTP. Both REMSD and 5-HT depletion via PCPA resulted in an increase in avoidance responding that was reversed by administration of 5-HTP. REMSD and 5-HT depletion via PCPA resulted in increased avoidance efficiency and were reversed by 5-HTP administration, but only changes following PCPA injection were statistically significant. Decreases in 5-HT levels that occur during REMSD likely mediate increases in avoidance responding. PMID- 12644282 TI - The dynamics of spine density changes. AB - Numerous papers have been published describing the effects of learning and environmental changes on the wiring of brain areas in mammals and birds. The density of dendritic spines, which can be taken as a measure of the complexity of a given neuronal network, has been shown to increase or to decrease depending on the experiment and on the brain area involved. Almost no information is available concerning the speed with which a given network reacts to learning events or environmental changes. We therefore examined the time course of spine density changes in two areas of the zebra finch forebrain, which have been shown previously to be either involved in sexual imprinting (LNH, lateral part of the neo-hyperstriatum) or to react to environmental changes (ANC, archi-neostriatum caudale). The decrease of spine density in LNH of zebra finch males after sexual imprinting is very fast, the new level of spine density is reached after 2 days. In contrast, decrease of spine density within ANC as a consequence of transferring birds from a social condition into isolation is very slow, lasting about 3 weeks. The increase of spine density within ANC after transfer of the males from isolation to a social condition occurs within 3 days. The differences in adaptation times cannot be due to limitations in the growth speed of single spines, because this has been shown to be much faster (hours instead of days). Instead, the speed of adaptation may be dependent on the availability of information about the final wiring diagram and on functional aspects like the energy demands for maintenance or alteration of a given neuronal network, or the necessity of quick adaptation to enhance the fitness of the animal. PMID- 12644283 TI - Inbred mouse strain differences in the establishment of long-term fear memory. AB - Studies describing variations in fear-related memory in inbred mouse strains typically focus upon 24 h retention. As a consequence, little is known about strain differences in the establishment of longer lasting memories of aversive events. In the present study, male mice from the strains A/Ibg, AKR/J, BALB/cByJ, CBA/J, C3H/HeIbg, C57BL/6J, DBA/2J, LP/J, SJL/J and 129/SvevTac were tested 24 h, 14, or 60 days after contextual and auditory-cued fear conditioning. Consistent with previous data, 24 h after conditioning these strains exhibited substantial variation in levels of memory for the context and the auditory cue as measured by freezing scores. Sixty days after training, most strains exhibited some forgetting of the context and auditory cue, and again there was significant strain variation. Strain rankings at 60-day retention were similar to that at 24 h with a significant genetic correlation between freezing values for the two time periods. Fourteen days following training, nearly all strains exhibited generalized freezing, a behavioral phenotype originally observed in C57BL/6 but not DBA/2 mice. These data confirm that cognitive differences exist between several popular inbred mouse strains during 24 h contextual fear recall. In addition, they extend these differences into retention time frames longer than those typically used and reveal several unique learning profiles of mouse strains that may be useful in furthering our understanding of how memories are formed. Emotionally arousing situations are often recalled a great deal of time after an event. Therefore, a more complete picture of the biochemical and genetic underpinnings of learning and memory will benefit from studies using time points that assess time points beyond 24 h retention. The utility of the 14-day hyper responsiveness phenotype as a potential model for fear-related psychopathology is also discussed. PMID- 12644284 TI - Testing the importance of the caudal retrosplenial cortex for spatial memory in rats. AB - Although there is evidence to suggest that the retrosplenial cortex is involved in spatial learning and memory, many lesion studies have left the more caudal part of this region intact so leaving its role untested. In the first experiment, rats with neurotoxic lesions of the caudal half of the retrosplenial cortex (RspC1) were tested on a reference memory task in the water-maze. The RspC1 animals were impaired on initial acquisition although they performed normally on a subsequent probe test. The second experiment looked at working memory in the radial-arm maze and water-maze. Animals with caudal retrosplenial lesions (RspC2) were unimpaired on the acquisition stage of the radial-arm maze task but were impaired when the task involved maze rotation to control for the possible use of intramaze cues. The RspC2 animals also took longer to learn the platform position on a delayed matching-to-place task in the water-maze. These results show a subtle impairment in spatial memory performance that is not as severe as that seen when more complete lesions of the retrosplenial cortex are made. PMID- 12644285 TI - Taste responsiveness and diet preference in autoimmune MRL mice. AB - One of the most profound behavioural deficits in lupus-prone MRL-lpr mice is blunted responsiveness to sweet solutions. Given the systemic nature of autoimmune/inflammatory disease, it was not clear whether impaired taste sensitivity or motivated response to palatable food underlie this deficit. The present study compares response rates of MRL-lpr mice (which develop disease early), congenic MRL +/+ mice (which develop disease later in life) and non autoimmune Swiss Webster (SW) mice to different tastes and diets. Healthy SW mice showed the highest responsiveness to palatable stimulation throughout the study. Conversely, the preference for palatable solutions progressively declined in MRL lpr mice as the disease developed. No differences between the two MRL substrains were seen in responsiveness to quinine or saline, suggesting that blunted responsiveness to palatable solutions cannot be accounted for by reduced taste sensory function (hypogeusia). In addition, changes in response rates to palatable solutions were associated with systemic upregulation of pro inflammatory cytokines. With a new cohort of mice fed on carbohydrate-rich and fat-rich diets, we also examined whether reduced sucrose intake in MRL-lpr mice can be accounted for by a reduced craving for carbohydrates. Contrary to this expectation, diseased MRL-lpr mice preferred carbohydrate-rich food while consuming a food mass comparable to controls. These results further support the hypothesis that the onset of lupus-like disease alters motivated behaviour, independent of changes in neurologic function and food metabolism. PMID- 12644286 TI - The effects of PACAP and PACAP antagonist on the neurobehavioral development of newborn rats. AB - Recent studies show that pituitary adenylate cyclase activating polypeptide (PACAP) plays an important role in the development of the nervous system. The aim of the present study was to investigate the effects of PACAP38 and the PACAP antagonist PACAP6-38 on the development of neonatal behavior in rats. Pups were treated subcutaneously until day 14, a period during which the blood-brain barrier is not yet complete. Rats were tested daily for the appearance of physical features, sensory and motor neurological signs, and for exploratory behavior on days 14 and 21. Facial development and most neurological signs were accelerated by PACAP treatment, while anti-PACAP retarded ear unfolding, eye opening, hindlimb placing and righting reflex. PACAP-treated animals also showed altered behavior in the open-field, in particular at 3 weeks of age. The number of areas entered and rearings were much higher than in the vehicle-treated group, and they spent less time along the walls and in corners. Anti-PACAP had little effect in the exploratory behavior of the pups. In summary, these data provide additional evidence for the neurotrophic effects of both endogenously present and exogenously administered PACAP-38. PMID- 12644287 TI - Antidepressant- and anxiolytic-like effects of selective neuronal NOS inhibitor 1 (2-trifluoromethylphenyl)-imidazole in mice. AB - Various inhibitors of nitric oxide synthase (NOS) have been shown to possess antidepressant- and anxiolytic-like properties in animal models. The aim of this study was to compare the behavioural effects of NOS inhibitor 7-nitroindazole (7 NI) with the more selective neuronal NOS inhibitor 1-(2 trifluoromethylphenyl)imidazole (TRIM) in animal models predictive of antidepressant- and anxiolytic-like activity in order to clarify the role of distinct isoforms of NOS in the regulation of depression and anxiety. Both TRIM (50 mg/kg) and 7-NI (50 mg/kg) decreased the immobility time in the forced swimming test. The magnitude of the effect was comparable to that of the tricyclic antidepressant imipramine (15 mg/kg). The antidepressant-like effect of TRIM was counteracted by pretreatment with L-arginine (250 mg/kg). The systemic administration of TRIM (50 mg/kg), but not 7-NI (up to 50 mg/kg) increased the time spent in the light side of the apparatus in the light-dark compartment test. The anxiolytic-like effect of TRIM was antagonised by pretreatment with L arginine. Both TRIM and 7-NI decreased the locomotion of animals in the open field and caused motor incoordination on rotarod. These motor side effects were more pronounced in the case of 7-NI and were not diminished by pretreatment with L-arginine. We conclude that neuronal NOS seems to play the key role in the antidepressant- and anxiolytic-like effects of NOS inhibitors. PMID- 12644288 TI - Covert muscle excitation is outflow from the central generation of motor imagery. AB - Two studies were conducted in an attempt to examine inflow and outflow processing by examining covert muscle excitation during motor imagery (MI) and its correlation with motor task performance. Examining 80 novice dart throwers in Experiment 1, MI produced greater levels of covert excitation at the dominant biceps in comparison to control imagery (CI). In addition, covert excitation correlated significantly with imagery ability and imagery vividness. This excitation, however, did not predict motor task acquisition or retention. Experiment 2 attempted to manipulate pre-imagery relaxation states by giving 104 novice dart performers a relaxation task or a distraction task before imagery sessions. MI resulted in improved task retention and resulted in significantly greater covert muscle excitation at the frontalis in comparison to CI. The relaxation condition, however, did not yield greater levels of muscular relaxation in comparison to the distraction condition, and did not yield greater imagery vividness or motor performance. Finally, covert muscle excitation did not predict motor acquisition or retention error. Results suggest that covert excitation is a byproduct of the central generation of the image that does not relate meaningfully to motor skill acquisition or retention gains. PMID- 12644289 TI - Effects of electrical stimulation or lesion in nucleus accumbens on the behaviour of rats in a T-maze after administration of 8-OH-DPAT or vehicle. AB - Electrical brain stimulation may be a therapeutic alternative for irreversible lesions in treatment-resistant patients with obsessive-compulsive disorder (OCD). We compared the effects of electrical stimulation and lesion in the nucleus accumbens (n acc) on the behaviour of rats in a model for OCD. Rats were tested for spontaneous alternation behaviour (AB) in a T-maze and assigned to four groups: an electrode implant group with stimulation 'ON' (stimON) or 'OFF' (stimOFF), a lesion or a sham group. Postoperatively, the number of arm visits and AB were tested after 8-hydroxy-2-(di-n-propylamino)-tetralin hydrobromide (8 OH-DPAT; 2 mg/kg) or saline administration. After 8-OH-DPAT administration, more arm visits were counted in the stimON (92.2%) and lesion groups (79.3%) than in both control groups (stimOFF 54.2; sham 61.2%). AB was significantly decreased in the stimON (10.5%) and lesion groups (10.2%) relative to the sham (22.0%) but not to the stimOFF group (14.7%). After saline administration, rats performed more arm visits in the stimON (81.5% non-significant) and lesion groups (93.6% significant) relative to the stimOFF (70.8%) and the sham groups (74.5%). No significant differences, however, were observed for AB. In conclusion, both treatments resulted in a decreased AB after 8-OH-DPAT administration (modelling an increase in compulsions) and more arm visits. PMID- 12644290 TI - Spatial memory performance of freely-moving squirrel monkeys. AB - Few experiments have addressed the problem of cognitive map formation in non human primates. Therefore, a paradigm was developed to assess spatial memory formation in squirrel monkeys (Saimiri sciureus) moving freely in three dimensions. While moving on the walls and floor of a large test chamber, the animals learned to collect pieces of cereal from baited food-ports interspersed among non-baited ports. The cereal-pellets were not visible to the monkeys, so the animals needed to develop spatial memory to visit only the baited ports for food and avoid the non-baited ones. A session consisted of ten consecutive trials, and 3 successive sessions were conducted on each day for a 5-day period. For each trial, correct choices (CC; number of visited baited-ports) and incorrect choices (IC; number of visited non-baited ports) were registered, and spatial memory performance index (SMPI; ranging from 0.00 to 1.00) was calculated as follows: SMPI=(CC-IC)/CC. For each session, mean SMPI, session duration, total reaches into the non-baited ports, and total reaches into the baited ports were documented. In an 8-port task, where 4 food-ports were baited and 4 were non baited, the mean SMPI was higher than 0 in the first session (day 1), indicating the development of short-term spatial memory. By the fifth session (day 2), this index was significantly higher than in the first session, indicating the build-up of long-term spatial memory. These changes were related to a significant decrease in the total reaches into the non-baited ports. At the same time, the duration of the sessions and the total reaches into the baited ports did not change significantly. This paradigm can be used for (1) studying cognitive map formation in primates, (2) examining the underlying cellular and molecular mechanisms in integrative neurobiological experiments, and (3) screening cognition-enhancer drugs in a monkey model. PMID- 12644291 TI - Effects of adrenal medulla graft on recovery of GABAergic and dopaminergic neuron deficits in mice: behavioural, pharmacological and immunohistochemical study. AB - We studied the capacity of adrenal medullary transplant to restore the deficits of GABAergic and dopaminergic neurons in mice injected with quinolinic acid (QA), using an open field test as well as pharmacological and immunohistochemical techniques. We analysed behavioural traits-total locomotor activity, peripheral and central activities, grooming, leaning and rearing in the QA-lesioned mice and mice that had undergone adrenal medulla (AM) transplantation. We found that the adrenal transplant recovered a loss of GABAergic neurons. It reduced QA-induced hyperactivity in locomotion and improved emotional indices. In addition, immunohistochemical studies of catecholaminergic markers-tyrosine hydroxylase (TH), dopamine (DA) and neuronal vesicular monoamine transporter type 2- and a single post-trial injection of tetrabenazine (TBZ; 5 mg/kg) indicated that catecholamines-synthesising chromaffin cells in the AM grafts were also involved in the beneficial effects. A likely interpretation of this behavioural pattern of results is that adrenal medullary transplants set into play an interaction between GABAergic and DAergic factors. Our results may contribute to the clarification of the beneficial effects of AM transplants in striatal function. PMID- 12644292 TI - Effects of inhaled toluene and 1,1,1-trichloroethane on seizures and death produced by N-methyl-D-aspartic acid in mice. AB - Evidence exists that some abused solvents have N-methyl-D-aspartic acid (NMDA) antagonist activity, although which of their effects may be related to this mechanism is not well understood. The effects of toluene and 1,1,1 trichloroethane (TCE) on NMDA-induced seizures in mice were studied using three experimental protocols: (a) animals injected i.p. with 120 or 170 mg/kg NMDA and immediately afterwards exposed to solvent vapors or air for 30 min (co-exposure protocol); (b) mice exposed for 30 min to solvent or air, then injected with NMDA and placed in the chamber for a second 30-min exposure (pre-exposure+co-exposure protocol); and (c) mice that inhaled 4000 ppm toluene or air for 30 min twice a day, 6 h apart, for 7 days, and were injected with 120 mg/kg NMDA immediately before a 30-min toluene exposure (repeated exposure protocol). When given acutely, toluene, but not TCE, produced concentration-dependent protection against NMDA-induced seizures. Higher concentrations of toluene were also effective against the lethal effects produced by 170 mg/kg NMDA. Clearer effects were seen when the pre-exposure+co-exposure protocol was followed. Under these conditions the IC(50) for toluene was 739 ppm (653-825) against seizure occurrence and 2127 ppm (1966-2288) against lethality. Repeated exposure to toluene did not result in tolerance to its anticonvulsant effects. These results are consistent with the in vitro effects described for toluene as a noncompetitive NMDA antagonist and as a compound that enhances GABAergic transmission. The lack of protective effects of TCE is not consistent with its in vitro actions. PMID- 12644293 TI - Anxiolytic-like effects of 5-HT2 ligands on three mouse models of anxiety. AB - The behavioural effects of 5-HT(2) receptor agonists, 5-HT(2A) and 5-HT(2C) receptor antagonists were investigated in the mouse four plates test (FPT), light/dark paradigm (L/D) and the elevated plus maze (EPM), in order to elucidate the role of the 5-HT(2) receptor subtypes in these models and to address the inconclusive results previously reported using rat psychopharmacological models. All compounds were administered intraperitoneally 30 min before each test. DOI, a preferential 5-HT(2A) agonist (0.5-8 mg/kg) and BW 723C86, a 5-HT(2B) agonist (8 and 16 mg/kg) provoked an anxiolytic-like response in the FPT. In the EPM, an anxiolytic-like effect was observed for DOI (0.5, 1 and 2 mg/kg), BW 723C86 (0.5, 4, 8 and 16 mg/kg), RO 60-0175 a 5-HT(2C) agonist (4 mg/kg) and the non-selective 5-HT(2) receptor agonist mCPP (0.25 mg/kg.). Ketanserin, a 5-HT(2A/2C) non selective receptor antagonist (0.015 and 0.03 mg/kg), induced an anxiogenic-like effect in the L/D paradigm. The 5-HT(2C) antagonists (RS 10-2221, SDZ SER082 and SB 206553) were without effect in all three tests. These behavioural results are indicative of an anxiolytic-like action of 5-HT(2) receptor agonists, an anxiogenic-like effect of 5-HT(2A) receptor antagonism, whereas the blockade of 5 HT(2C) receptors are without effect in the mouse models studied. PMID- 12644294 TI - A new subgroup of the family 2 cystatins. AB - The cystatins are a superfamily of cysteine protease inhibitors. Several genes including Cres (cystatin-related epididymal spermatogenic), testatin, and cystatin T, have been identified that are related to the family 2 cystatins but lack critical consensus sites important for cysteine protease inhibition. In addition, these genes are primarily expressed in the reproductive tract suggesting they may have evolved to perform tissue-specific functions distinct from that of the typical cystatins. This review describes the CRES subgroup within the family 2 cystatins including potential new members and their putative functions in the reproductive tract. PMID- 12644295 TI - TSPY-LTA transgenic mice develop endocrine tumors of the pituitary and adrenal gland. AB - In an attempt to determine the susceptibility of spermatogonia to malignant transformation transgenic mice were generated harboring a 1.3 kb 5'-flanking region of the germ cell specific expressed human testis specific protein, Y encoded gene fused with the simian virus 40 large T antigen (TAg). Unexpectedly, TAg expression in transgenic mice was also detected in somatic tissues. Between days 65 and 85 after birth most of the transgenic mice developed anterior lobe tumors of the pituitary gland and to a less extent medulla type tumors of the adrenal gland. In addition, a few older transgenic mice developed tumors of the seminal vesicle, but no testicular tumors were observed in transgenic mice up to an age of 5 months. The pituitary tumors were immunoreactive for anti-prolactin (PRL) and anti-adrenocorticotropic hormone (ACTH). PRL and corticosterone concentrations in serum of transgenic mice were significantly increased. Taken together, our studies provide a novel mouse model for pituitary adenomas displaying a unique combination of hormone expression by tumor cells secreting PRL and ACTH. PMID- 12644296 TI - IGF-1 receptor mediates differentiation of primary cultures of mouse skeletal myoblasts. AB - Studies involving immortalized myoblasts suggested that insulin-like growth factors (IGFs) promote differentiation of skeletal muscle, but gene targeting experiments in mice did not provide support for this hypothesis. To address this discrepancy, we examined differentiation of primary cultures of mouse myoblasts. Differentiation was normally unaffected by addition of IGFs to the differentiation medium. However, when we interrupted IGF-mediated signaling, by incubating myoblasts with suramin or with a monoclonal antibody to the IGF-I receptor, differentiation was inhibited. Inhibition was reversed by exogenous IGF I or IGF-II, but not by insulin. Differentiation was enhanced in myoblasts that were incubated with an inhibitor of the mitogen-activated protein kinase signaling pathway (PD098059) and such cells were responsive to exogenous IGF-I. Our results demonstrate that IGF action contributes to the differentiation of non immortalized mouse myoblasts and that these cells represent a model system that can be experimentally manipulated to study the molecular events involved in skeletal muscle differentiation. PMID- 12644297 TI - Changes in oxytocin receptor in bovine preovulatory follicles between the gonadotropin surge and ovulation. AB - In cattle, production of oxytocin by granulosa cells of preovulatory follicles is induced by the LH/FSH surge and intrafollicular oxytocin increases dramatically toward the end of the interval between the surge and ovulation. We reported previously that oxytocin modulates steroid production by both theca and granulosa cells obtained from bovine preovulatory follicles, implying actions of oxytocin on both cell types of preovulatory follicles. The objective of the present study was to examine the temporal expression of oxytocin receptor mRNA and protein in both theca and granulosa cells of bovine periovulatory follicles. To induce luteal regression and initiate a follicular phase, heifers were injected with prostaglandin F(2alpha) on Day 6 or 7 of the estrous cycle and 36 h later, a GnRH analogue was administered to induce the LH/FSH surge. The periovulatory follicle was isolated at 0, 3.5, 12, or 24 h after GnRH injection. A significant increase in the levels of mRNA for oxytocin was detected in granulosa, but not theca, cells of periovulatory follicles at 12 and 24 h after GnRH injection, relative to time 0. In contrast, the levels of oxytocin receptor mRNA and specific binding sites for oxytocin in granulosa cells had decreased significantly at 12 and 24 h post-GnRH. In theca cells, the levels of oxytocin receptor mRNA were significantly lower at 12 and 24 h compared with values at 3.5 h, but specific binding of oxytocin to thecal cell membranes was not different at any time point. Immunopositive staining for oxytocin receptor was localized to both the theca and granulosa cell layer of periovulatory follicles at all four times of follicle isolation. These results suggest the direct action of oxytocin on both theca and granulosa cells of bovine periovulatory follicles through binding to its receptor, supporting the hypothesis that follicular oxytocin plays an important role(s) in the regulation of the final stage of follicular development. Down regulation of oxytocin receptor mRNA and oxytocin binding may serve to temporally limit the actions of oxytocin on the preovulatory follicle. PMID- 12644298 TI - Interdomain interactions in the mineralocorticoid receptor. AB - The potential for interaction between the N-terminal domain and the C-terminal region (hinge and ligand-binding domain) of the mineralocorticoid receptor (MR) was examined using the mammalian-2-hybrid assay. The MR C-terminal region was fused to the GAL4 DNA-binding domain (GAL4-MRC). To examine if the AF-2 is involved in the interaction, as has been reported for other steroid hormone receptors, it was inactivated by point mutation (E962A). The N-terminal domain was fused to the VP16 transactivation domain (VP16-MRNT). In the mammalian-2 hybrid assay both GAL4-MRC and GAL4-MRC(E962A) interact with VP16-MRNT in an aldosterone-dependent manner. The GAL4-MRC(E962A) construct was used in subsequent experiments to examine the AF-2-independent N/C-interaction. The MR antagonist spironolactone inhibits the aldosterone-mediated association of the two domains. GAL4-MRC(E962A) interacts weakly with the GR or AR N-terminal domains in the presence of aldosterone. No dimerization between GAL4-MRC(E962A) and VP16-MRC is observed. Interestingly, cortisol produces a much weaker N/C interaction than aldosterone, and it is possible that the N/C-interaction may contribute to observed functional differences in the MR bound to the two ligands. PMID- 12644299 TI - Circadian expression of Mel1a and PL-II genes in placenta: effects of melatonin on the PL-II gene expression in the rat placenta. AB - In the mammal, melatonin regulates the seasonal and/or circadian rhythm of PRL levels. Since several members of the PRL gene family are expressed during late pregnancy, we investigated the relationship between the expression of placental lactogen (PL)-II-one member of the PRL family- and melatonin, as well as the placental expression of one of the receptors for melatonin, melatonin receptor 1a (Mel(1a())). Herein we provide the first demonstration that Mel(1a) is not only expressed in the rat placenta, but that it is spatially and temporally regulated throughout late pregnancy. In situ hybridization and Northern blot analyses show that Mel(1a) mRNA is localized in the rat placenta on gestational day 19, and is mainly restricted to the spongiotrophoblast and trophoblast giant cells. Interestingly, the junctional zone of the placenta at this time showed the strongest gene expression when the tissue was obtained at 16:00 h (daytime) and showed the least expression when it was obtained at 04:00 h (night-time). In contrast, the labyrinth zone showed the strongest expression in tissue obtained at night and showed the least expression in tissue obtained during the day. PL-II gene expression also exhibited a circadian rhythm but the direction of the fluctuation was exactly opposite to that of the Mel(1a) gene, such that at night the junctional zone had the strongest expression, while the labyrinth zone had the weakest. In vitro treatment of placental tissue with an melatonin agonist, chloromelatonin, greatly decreased PL-II mRNA levels. That Mel(1a) plays a regulatory role in the expression of PL-II in the late-pregnancy rat placenta is strongly suggested by the pattern of its own spatial and temporal expression. PMID- 12644300 TI - Characterization of mammary tumor cell lines from wild type and vitamin D3 receptor knockout mice. AB - 1,25-Dihydroxyvitamin D(3) (1,25D(3)), the active metabolite of vitamin D(3), inhibits breast cancer cell growth in vivo and in vitro. To examine mechanisms of 1,25D(3) induced growth arrest and apoptosis, cell lines were established from DMBA induced mammary tumors derived from vitamin D(3) receptor knockout (VDRKO) and wild type (WT) mice. Two VDRKO (KO240, KO288) and two WT (WT145, WT276) cell lines were selected and characterized. All four cell lines express cytokeratins indicative of an epithelial origin, as well as vimentin, which is expressed in many transformed cell lines. The tumorigenicity of the cells was confirmed in vivo as all four cell lines form estrogen responsive tumors in nude mice. Both WT cell lines express the VDR protein and are sensitive to growth inhibition by 1,25D(3) at doses as low as 1 nM. Flow cytometric analysis indicated that 1,25D(3) induces G(0)/G(1) arrest and apoptosis in the WT cell lines. In contrast, both cell lines established from tumors that developed in VDRKO mice lack VDR mRNA and protein. Cells from WT mice exhibit 1,25D(3) inducible transcriptional activity, as measured by reporter gene assays, but cells from VDRKO mice do not. Cells from VDRKO mice are also completely resistant to 1,25D(3) mediated growth arrest and apoptosis over the range of 0.01-100 nM 1,25D(3). VDRKO cells are also resistant to the synthetic vitamin D(3) analogs EB1089 and CB1093 that are more potent growth inhibitors than 1,25D(3) in WT cells. This data conclusively demonstrate that the induction of cell cycle arrest and apoptosis in breast cancer cells by 1,25D(3), EB1089 and CB1093 is dependent on the nuclear VDR. Cells lacking VDR remain sensitive to growth arrest mediated by 9-cis retinoic acid, a ligand for the retinoid x receptor which can heterodimerize with the VDR. Sensitivity to apoptosis induced by the DNA damaging agent etoposide is not altered in VDRKO cells, indicating that VDR ablation does not impair apoptotic pathways in general. All four cell lines display equal sensitivity to tamoxifen induced growth arrest. These estrogen responsive, transformed cell lines which differentially express the VDR provide a novel model system for identification of the mechanisms by which 1,25D(3) regulates proliferation and apoptosis in breast cancer cells. PMID- 12644301 TI - Interferon-inducible genes in the rat adrenal gland and vascular smooth muscle cells. AB - Chronic stimulation of the renin-angiotensin system results in increased zona glomerulosa cells and in cells expressing the final enzyme in the synthesis of aldosterone, the cytochrome P-450 aldosterone synthase. The genes activated during adrenal remodeling are not well defined. We have reported that the expression of interferon-inducible genes, 9-27, 1-8D and 1-8U in H295R cells is stimulated by A-II. The 9-27 gene is expressed mainly in leukocytes and is associated with cell proliferation. In this study, we searched for similar genes in a rat zona glomerulosa cDNA library, and examined the regulation of the expression of these genes. We found the Rat8 gene, which has been reported to be similar to human interferon-inducible genes, as well as two similar genes, No. 10 (1096 bp), and No. 16 (630 bp). Rat8 gene and No. 16 were mainly expressed in zona glomerulosa. The product of No. 10 is thought to be a secreted protein, unlike those of 8 and 16, and its expression in the adrenal was weak in comparison. The control of the expression of rat8 or No. 16 genes differs depending on the tissue. Expression in A10 cells (derived from rat embryo thoracic aorta) was not stimulated by A-II, nor was it influenced by salt intake in the adrenal gland, but it was reduced in vascular smooth muscle cells (VSMC) of rats on a low sodium diet. These results show that genes similar to the human 1-8 gene family are expressed in rat adrenal glomerulosa cells and VSMC, but their expression is not regulated by A-II. The function of these genes in VSMC and adrenal is unknown. PMID- 12644302 TI - Transcriptional regulation of the GnRH receptor gene by glucocorticoids. AB - Expression of the gonadotropin-releasing hormone (GnRH) receptor gene is stimulated by dexamethasone in GnRH-deficient rodents. In this study we identify a 1226 bp sequence at the 5'-flanking region of the mouse GnRH-R gene that confers dexamethasone responsiveness when expressed in host cells. Further, a glucocorticoid antagonist blocks transcriptional activity of the mGnRHR promoter. Progressive 5'-deletion of the mGnRHR promoter localized the response sequence between the -331/-255 region. Analysis of this region revealed binding sites for the AP-1 transcription factor. Mutation in AP-1 modified the functional activity of the mGnRHR promoter following GnRH agonist or dexamethasone-stimulation. Using an electrophoretic mobility shift assay, a protein complex is shown to bind to the AP-1 site. These results suggest that AP-1 proteins and glucocorticoid receptor regulate transcription of the GnRH-R promoter in a heterologous system. PMID- 12644303 TI - ACTH and alpha-MSH inhibit leptin expression and secretion in 3T3-L1 adipocytes: model for a central-peripheral melanocortin-leptin pathway. AB - Leptin is the 167 amino-acid protein product of the Lep (obese) gene that is released predominantly from adipose tissue and circulates at levels related to the amount of fat. Leptin expression is hormonally regulated: insulin and glucocorticoids are stimulators, while inhibitors include beta-adrenergic agonists and testosterone. Recently, adenylate cyclase-coupled melanocortin receptors have been identified in murine adipose tissue, the 3T3-L1 adipocyte cell line, and in human fat tissue. These studies prompted us to evaluate the effects of pro-opiomelanocortin (POMC)-derived peptides on leptin production and expression in 3T3-L1 adipocytes in culture. 3T3-L1 pre-adipocytes differentiated by the insulin/indomethacin (I/I) method produced leptin at levels that were two times higher than those obtained in cells differentiated by the more traditional insulin/dexamethasone/isobutylmethylxanthine (I/D/M) method. By RT-PCR studies, 3T3-L1 cells expressed both the melanocortin 2 receptors (MC2-R) and melanocortin 5 receptors (MC5-R) isoforms of the melanocortin receptor at an early stage of differentiation. When I/I differentiated 3T3-L1 adipocytes were incubated with different concentrations of dibutyryl cAMP (db-cAMP) or POMC-derived peptides (ACTH and alpha-MSH), ACTH and alpha-MSH stimulated cAMP production after 30 min (2-fold increase) associated with a dose-dependent inhibition of leptin secretion (ACTHz.Gt;alpha-MSH; IC(50)=3.2+/-0.4 SE and 36+/-5 nM, respectively), maximal after 3 h of incubation (30% inhibition). In addition, 100 nM ACTH and alpha-MSH induced a 60% reduction in leptin expression by RT-PCR. Incubation of cells with 0.5 mM db-cAMP led to a more prominent inhibition of leptin expression and secretion (up to 80% at 1 and 24 h, respectively). The ACTH and alpha-MSH inhibitory effects on leptin secretion were mediated by activation of the MC2-R and MC5-R and were reversed by the MC-R antagonists ACTH(11-24) and ACTH(7-38). In summary, we have shown that POMC-peptides are potent inhibitors of leptin expression and production in 3T3-L1 adipocytes. The finding of ACTH/alpha-MSH receptor-induced inhibition of leptin production and expression in adipocytes support the possibility that there is a control mechanism for modulation of adipose tissue function via a melanocortin-leptin axis. PMID- 12644304 TI - Type 2 deiodinase expression is stimulated by growth factors in human vascular smooth muscle cells. AB - Type 2 deiodinase (D2) catalyzes the conversion of the prohormone T4 to the biologically active T3. D2 is expressed in human aortic smooth muscle cells (hASMCs). In this study, we demonstrated that the D2 mRNA and activity in hASMCs were up-regulated by platelet-derived growth factor-BB (PDGF-BB) and basic fibroblast growth factor (bFGF). The induction of D2 mRNA by PDGF-BB and bFGF was dependent on de novo RNA and protein synthesis. PD98059, a specific inhibitor of the upstream kinase that activates extracellular signal-regulated kinase (ERK), significantly suppressed the induction by both PDGF-BB and bFGF. SB203580, a specific inhibitor of p38 mitogen-activated protein (MAP) kinase, and SP600125, a specific inhibitor of c-Jun N-terminal kinase (JNK), also reduced the induction by both PDGF-BB and bFGF. These results suggest that both PDGF-BB and bFGF induce D2 expression at least partly via ERK pathway. The p38 MAP kinase and JNK pathways may also be involved in the induction. PMID- 12644305 TI - A Galphas mutation (D229S) differentially effects gonadotropin-releasing hormone receptor regulation by RGS10, RGS3 and RGS3T. AB - Regulators of G protein signaling (RGS) act as GTPase-activating proteins for Galpha(i) and for Galpha(q/11). There is recent evidence for interaction of RGS proteins with Galpha(s), and substitution of Ser for Asp(229) in RGS proteins enhances interactions with G proteins. Site-directed mutagenesis of Asp(229) was used to assess the effect of this site on the gonadotropin-releasing hormone receptor (GnRHR)-Galpha(s) mediated signaling in the absence or presence of over expressed RGS3, RGS10 or a truncated form of RGS3 (RGS3T). We observed increased cAMP release with the mutant Galpha(s)(D(229)S) compared to wt Galpha(s) when GGH(3) cells (GH(3) cells stably expressing the GnRH receptor) were stimulated with a GnRH agonist. In the presence of RGS3, we did not observe any difference in cAMP release with wt Galpha(s) or with Galpha(s)(D(229)S) compared to control values; in the presence of RGS3T there was an increase of cAMP release with wt Galpha(s) compared to the control but there was no difference between the Galpha(s)(D(229)S) and the control values. When cells co-expressed wt Galpha(s) and RGS10, there was a significant increase of cAMP release compared with cells co-expressing wt Galpha(s) and Lac Z. Cells co-expressing Galpha(s)(D(229)S) and RGS10 showed a significant increase of cAMP release compared to control cells. These results indicate differential regulation of the GnRHR-Galpha(s) mediated signaling by a single mutation in Galpha(s) in the presence of RGS10 and RGS3T, but not with RGS3. This is the first report of an effect of the Galpha(s)(D(229)S) mutation on G protein-coupled receptor-mediated activation. PMID- 12644306 TI - Evidence for differential ribonucleoprotein complex assembly in vitro on the 5' untranslated region of the human IGF-IR transcript. AB - The type I insulin-like growth factor receptor (IGF-IR) plays a key role in the control of cellular proliferation and survival. The human IGF-IR transcript is characterized by an unusually long 1038 nucleotide 5'-untranslated region (5' UTR). We hypothesized that the contribution of this complex 5'-untranslated RNA sequence to the post-transcriptional regulation of IGF-IR expression would involve a dynamic interplay between RNA structure and specific RNA-binding proteins. Here we have detected and characterized a diverse series of regulatory proteins binding the IGF-IR 5'-UTR under disparate conditions. One pair of proteins ( approximately 42/38 kDa) binds readily to the intact 5'-UTR, which is predicted to adopt a highly base-paired, highly favorable (dG=-498 kcal/mol) three-domain structure. Another protein(s) (p20*) specifically induces formation of a novel RNA structure from within the initial 209 nucleotides of the nascent IGF-IR transcript, but fails to UV crosslink to this RNA sequence. A third group of proteins recognizes and binds the IGF-IR 5'-UTR under highly stringent conditions, but only after higher-ordered RNA structure has been disrupted. Our in vitro results indicate that the IGF-IR 5'-UTR may exist in at least three distinct states, and we propose that interconversion between these states might take place in vivo and differentially alter IGF-IR transcript utilization. PMID- 12644308 TI - Cloning and expression of a novel, truncated, progesterone receptor. AB - Progesterone acts via two specific receptors to affect gene transcription in target tissues. Progesterone receptor (PR) B contains 933 amino acids while PR A is a truncated version lacking the initial 164 amino acids. We have cloned a novel, truncated PR from both human adipose and aortic cDNA libraries. This cDNA encodes a predicted protein of 314 amino acids, termed PR-M. Initiation of transcription of PR-M occurs in intron 3, with the initial exon identical to exon 4 of the genomic PRs, except for a novel 16 amino acid amino-terminal sequence, consistent with a signal peptide. The remainder of PR-M is identical to the genomic PR. Transcript for this protein was identified by RT-PCR in human aortic endothelial cells and T47D breast cancer cells. Expression of PR-M in Sf 9 insect cells results in a 38-kDa protein, demonstrated in human aortic endothelial cells and T47D breast cancer cells. The function of PR-M remains to be determined. The presence of a signal peptide and the lack of a DNA binding region suggests a non genomic action. PMID- 12644307 TI - Epidermal growth factor induces c-fos and c-jun mRNA via Raf-1/MEK1/ERK-dependent and -independent pathways in bovine luteal cells. AB - Epidermal growth factor (EGF) modulates the actions of gonadotropins in the corpus luteum. The membrane-associated EGF receptors undergo rapid tyrosine phosphorylation and internalization upon ligand binding in ovarian cells, including luteal cells. However, little is known about the post-receptor signaling events induced by EGF that lead to the transcriptional regulation of EGF-responsive genes in the ovary. The present study was designed to examine in bovine luteal cells (1) activation of the extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase (MAPK) signaling cascade (Raf/MEK/ERK) by EGF; (2) mRNA expression of AP-1 transcription factors, i.e. c-fos and c-jun, in response to EGF; and (3) the role of ERK in EGF-induced expression of c-fos and c jun mRNA. Raf-1 and B-Raf, but not A-Raf, were activated by EGF (10 ng/ml) and the pharmacological protein kinase C (PKC) activator phorbol myristate acetate (PMA, 20 nM). Activation of Raf resulted in the phosphorylation and activation of MAPK kinase (MEK1) which subsequently activated ERKs. Treatment with EGF-induced the phosphorylation of both ERK2 and ERK1 in a time and concentration dependent manner. Additionally, activated ERK was found in the nucleus of the cells following treatment with EGF (10 ng/ml) and PMA (PMA, 20 nM) for 5 min. Depletion of PKC by chronic PMA treatment (2.5 microM, 24 h) only partially inhibited the stimulatory effects of EGF on Raf-1, ERK2 and ERK1. These data demonstrate that PKC-dependent and independent-mechanisms are involved in EGF activation of the Raf/MEK/ERK signaling cascade in bovine luteal cells. EGF rapidly and transiently stimulated the expression of c-fos and c-jun mRNA in bovine luteal cells. Maximal induction of c-fos and c-jun mRNA by EGF occurred within 30 min of treatment with 10 ng/ml EGF. Treatment with the MEK1 inhibitor PD098059 (50 microM) abolished EGF-induced ERK activation. However, blocking EGF-induced ERK activation by pretreatment with PD098059 only partially attenuated EGF-induced c-fos and c-jun mRNA expression. Thus, additional pathways are implicated in the regulation of c fos and c-jun mRNA expression by EGF in bovine luteal cells. PMID- 12644309 TI - Adenoviral-mediated over-expression of Brn2 in the rat paraventricular nucleus: no effect on vasopressin or corticotrophin releasing factor RNA levels. AB - We have used an over-expression strategy to test the hypothesis that the Class III POU transcription factor Brn2 is rate limiting in the control of the level of expression of the vasopressin (VP) gene in the paraventricular nucleus of the rat hypothalamus. Knockout studies in mice have suggested that Brn2 may contribute to the control of the level of VP gene expression in the adult hypothalamus. However, we show here that in heterologous cell lines, Brn2 transactivates neither the proximal promoter of the rat VP gene, nor a novel reporter construct consisting of the rat VP structural gene and 3 and 2 kbp of upstream and downstream flanking sequences. We hypothesised that this maybe due either to the lack of cis-acting elements within the confines of the reporter vectors used, or to the absence in heterologous cells, of factors required for Brn2 activity. As no cell lines exist that correspond to VP neurons, we devised an adenoviral vector delivery strategy that enabled efficient over-expression of Brn2 in the paraventricular nucleus of the intact rat. Localised over-expression of Brn2 had no effect on VP hnRNA levels. Neither did we detect corticotrophin releasing factor (CRF) mRNA up-regulation by Brn2 over-expression in vivo. This was unexpected as Brn2 transactivates the proximal CRF promoter in vitro. Whilst Brn2 is required for the development of the hypothalamic structures that express VP and CRF, these data suggest that this transcription factor is not required, or is not rate limiting, for expression in the adult. PMID- 12644310 TI - Characterization of complementary deoxyribonucleic acids encoding preprogalanin and its alternative splice variants in the goldfish. AB - This paper reports the identification of five preprogalanin complementary deoxyribonucleic acids encoding preprogalanin peptides in the goldfish. Preprogalanin 1A, 1B and 1C are encoded in galanin gene 1 and 2A and 2B are encoded in gene 2. Preprogalanin 1B and 2B have a 24 amino acids insert in the mature peptide region and form 1C has a deletion of 23 amino acids in the middle of the galanin message associated peptide region. The mature peptides from the preprogalanin 1A and 1C are 29 amino acids. However, the mature peptide is 31 amino acids from preprogalanin 2A, 53 amino acids from 1B and 55 amino acids from 2B. The physiological significance of multiple forms of galanin peptide is unknown. Organization of galanin gene 1, which is similar to the mammalian galanin gene has been identified. Expression of preprogalanin messenger ribonucleic acids was widely detected in goldfish brain and several peripheral tissues. PMID- 12644311 TI - Activation of the MAP kinase cascade by exogenous calcium-sensing receptor. AB - In Rat-1 fibroblasts and ovarian surface epithelial cells, extracellular calcium induces a proliferative response which appears to be mediated by the G-protein coupled calcium-sensing receptor (CaR), as expression of the nonfunctional CaR R795W mutant inhibits both thymidine incorporation and activation of the extracellular-regulated kinase (ERK) in response to calcium. In this report we utilized CaR-transfected HEK293 cells to demonstrate that functional CaR is necessary and sufficient for calcium-induced ERK activation. CaR-dependent ERK activation was blocked by co-expression of the Ras dominant-negative mutant, Ras N17, and by exposure to the phosphatidyl inositol 3' kinase inhibitors wortmannin and LY294002. In contrast to Rat-1 fibroblasts, CaR-mediated in vitro kinase activity of ERK2 was unaffected by tyrosine kinase inhibitor herbimycin in CaR transfected HEK293 cells. These results suggest that usage of distinct pathways downstream of the CaR varies in a cell-type specific manner, suggesting a potential mechanism by which activation of the CaR could couple to distinct calcium-dependent responses. PMID- 12644312 TI - The phosphatidylinositol 3-kinase inhibitor LY294002 binds the estrogen receptor and inhibits 17beta-estradiol-induced transcriptional activity of an estrogen sensitive reporter gene. AB - Estrogen receptors (ERs) are members of the superfamily of ligand-activated transcription factors. In addition to the classical, hormone-mediated activation, ERs may alternatively be activated in a ligand-independent manner by a variety of agents including growth factors, neurotransmitters and cAMP. It has been demonstrated that the phosphatidylinositol 3 (PI3)-dependent kinase/Akt pathway may activate the ER alpha by increasing the activity of both estrogen independent activation function-1 and estrogen-dependent activation function-2 domains. The Akt phosphorylation site in the ER is Ser167. Phosphorylation of this residue is inhibited by LY294002, which blocks the PI3-kinase/Akt pathway. In the course of studies examining the effects of LY294002 on ligand-independent activation of ERs in L cells, we found that LY294002 exhibits antiestrogenic effects in a dose dependent manner. By competition binding assays, we found that LY294002 specifically displaced radiolabelled estradiol from ERs with an IC(50) of 11+/ 0.06 nM, being an estradiol competitor as effective as the antiestrogens ICI182,780 (IC(50), 21+/-0.13) and 4-OH-tamoxifen (IC(50), 15+/-0.09). Further, LY294002 irreversibly blocked estrogen-induced transactivation of an estradiol sensitive reporter gene. These findings are of particular importance in the interpretation of studies demonstrating ERs inactivation by the PI3-kinase inhibitor. Our studies show that an apparent block of ER activation cannot be dissociated from inhibition of ligand-mediated events. Thus, this effect can be the result of the ability of LY294002 to bind the ERs and inhibit transactivation of estrogen-regulated genes. PMID- 12644313 TI - IL-10, an inflammatory/inhibitory cytokine, but not always. AB - IL-10 has been previously called cytokine synthesis inhibiting factor, produced mostly by Th2 cells, macrophages and CD8+ cell clones. IL-10 is capable of inhibiting the synthesis of several cytokines from different cells, antigen or mitogen activated. IL-10 exerts its inhibition at the mRNA transcriptional and translational level. In addition, IL-10 is a co-stimulatory cytokine on activated T cells. For example, IL-10 inhibits NK cell activity, the production of Th1 cytokines, cytokines generated by peripheral blood mononuclear cells, and macrophage activity. On the other hand, IL-10 exerts immunostimulatory effects on B cells, cytotoxic T cell development and thymocytes. In mast cells derived from CD4+/CD133+ cells, IL-10 inhibits IL-6 and TNFalpha, and prostaglandin E(1) and E(2) induced by IL-6. Here, we report for the first time that IL-10 fails to inhibit tryptase and IL-6 from human mast cell-1 (HMC-1) and human umbilical cord blood-derived mast cells. PMID- 12644314 TI - Infection pattern and immune response in the spleen and liver of BALB/c mice intracardially infected with Leishmania donovani amastigotes. AB - Intracardial inoculation of BALB/c mice with Leishmania donovani amastigotes induced progressive visceral leishmaniasis (VL) with increasing splenic parasite load when followed upto 4-month postinfection period. In contrast, the liver parasite load reached maximum around 2-month postinfection period following which it started declining. The infection pattern differed somewhat from the earlier reports on mouse model of VL induced by intravenous inoculation of parasites with respect to the duration as well as magnitude of parasite burden in the organs (liver and spleen) and associated hepatosplenomegaly. Immunosuppression in mice with progressive VL was manifested in the form of impairment of proliferative response of the splenic mononuclear cells (SPMC) to in vitro stimulation with leishmanial antigen or the mitogen concanavalin A (ConA), although ConA stimulated cells were found to be capable of IL-2 and IFN-gamma synthesis. Differential expression of activating (IL-2, IFN-gamma and TNF-alpha) as well as deactivating (IL-4 and TGF-beta) cytokines was demonstrable in the spleen and liver of animals during the course of infection. Further, the synthesis of inducible nitric oxide synthase (iNOS) enzyme increased considerably in the liver as well as in the spleen of 4-month infected animal with parallel increase in the transcripts of the iNOS activating cytokines IFN-gamma and TNF-alpha. The temporal variation in the organ specific immune response could be related to the differential control of parasite burden in the liver and spleen of the infected host. PMID- 12644315 TI - Exposure to fatty acids modulates interferon production by intraepithelial lymphocytes. AB - Intraepithelial lymphocytes (IELs) play important roles in intestinal mucosal immunity. Although fatty acids are known to modulate the functions of immune effector cells, there has been no information about how fat exposure affects immunological function of IELs. In this study, we examined how fatty acids of various chain lengths modulate the production of interferon (IFN)-gamma by IELs stimulated with T-cell receptor (TCR) or interleukin (IL)-12/IL-18. IELs isolated from the small intestine of BALB/c mice were stimulated with plate-coated anti CD3 monoclonal antibody (mAb) or IL-12/IL-18. They were coincubated in microtiter plates for 3 days with various concentrations of fatty acid micelles. We used arachidonic acid, linoleic acid, and oleic acid as long-chain fatty acids, and used octanoic acid as a medium-chain fatty acid. IFN-gamma in the supernatants were measured by ELISA, and the expression of IFN-gamma mRNA in IELs was determined by RT-PCR. Significant production of IFN-gamma from IELs was observed after anti-CD3 mAb stimulation. The combination of IL-12 and IL-18 induced significant levels of IFN-gamma production without TCR stimulation. Increased IFN gamma mRNA was also observed after anti-CD3 or IL-12/IL-18 stimulation. Long chain fatty acids dose-dependently inhibited the stimulated-IFN-gamma production at concentrations greater than 10 micro M, but the medium-chain fatty acid did not cause any significant changes in IFN-gamma production. IFN-gamma production from gammadelta IELs was very low compared with alphabeta IELs, however, both populations showed similar attenuating patterns when treated with long-chain fatty acids. There is a possibility that the exposure of IELs to intraluminal fatty acids significantly modifies the immune function of intestinal mucosa. PMID- 12644316 TI - Skewed inhibitory receptors expression in a TAP2-deficient patient. AB - Most of patients suffering from HLA class I deficiency due to mutations in TAP genes show a significative increase of the peripheral minor Vdelta1+ subpopulation of gammadelta T cells. Surface expression of inhibitory receptors (IR) for HLA class I molecules have been mainly attributed to Vdelta2+ gammadelta T clones. In this study we have analysed the expression of these receptors in both subsets of gammadelta T peripheral lymphocytes. We studied 16 healthy controls and a reported case of homozygous TAP2 mutation with a marked increase of Vdelta1+ gammadelta T cells. MICA/B presence in monocytes was also evaluated. In healthy subjects, the expression of CD94 and CD94/NKG2A was higher in the Vdelta2+ subset but cells bearing the IR ILT2 were found increased in the Vdelta1+. The patient Vdelta2+ gammadelta T cells showed the same IR expression than normal controls, in contrast the Vdelta1+ subset presented a special pattern of very high expression of CD94 and ILT2 and low of CD94/NKG2A. The presence of a new IR poorly represented in healthy individuals could account for the selective increase of Vdelta1+ gammadelta T in TAP-deficient patients. MICA/B surface expression in monocytes was not shifted in our patient. PMID- 12644318 TI - IL-7 receptor is present on human microvascular endothelial cells. AB - Interleukin-7 (IL-7) is a pleiotropic, non-redundant cytokine crucial for development of B and T lymphocytes. The cellular response to IL-7 is triggered by binding of the cytokine to its receptor, IL-7R. Until now the expression of the receptor was evidenced only in lymphoid and myeloid cell lineages. The receptor consists of two chains: IL-7 specific alpha chain (CD127) and the common gamma(c) chain (CD132) which is a component of several other cytokine receptors: IL-2, IL 4, IL-9 and IL-15. The former observation that exogenous IL-7 is biologically active towards murine endothelial cell lines from secondary lymphoid organs was the starting point of our studies. This observation has prompted us to search for the presence of IL-7 receptor in human microvascular endothelial cells. We used in our studies a set of human endothelial cell lines established from various organs. Our results demonstrate the presence of IL-7R in human microvascular endothelial cells, mainly in the mesenteric but also in the peripheral and to a lesser extent, in the mucosa-associated lymph node endothelial cells. On the basis of the RT-PCR reaction, molecular weight estimated in Western blot and IL-7 binding activity the identified endothelial IL-7 receptor was identical to the lymphocyte-type IL7-R. PMID- 12644317 TI - Identification of antibodies to Leishmania silent information regulatory 2 (SIR2) protein homologue during canine natural infections: pathological implications. AB - Dogs are the domestic reservoir of zoonotic visceral Leishmaniasis caused by Leishmania infantum in the Mediterranean basin and thus constitute an important health problem in both human and veterinary medicine. Until vaccines become available, conventional measures such as epidemiological surveillance including reservoir control will be among the practical options for prevention and containment of the disease. We have recently characterised novel Leishmania sp. genes encoding parasite proteins named (LmS3a: homologous to mammalian ribosomal protein S3a; LmSIR2: homologous to the silent information regulatory 2 protein family; LimTXNPx: homologous to the peroxiredoxin family with N-terminal mitochondrial leader sequence) that may contribute to the host immune dysfunction in murine experimental Leishmaniasis. In the present study we have investigated the humoral responses against the parasite antigens in groups of L. infantum infected dogs with different clinical status: symptomatic and asymptomatic with DTH positive or negative test. The determination of immunoglobulin (Ig) isotypes revealed high levels of total IgG in both symptomatic and asymptomatic animals when compared to IgM. Furthermore, the IgG2 appeared to be the predominant subclass of Ig present in the sera of infected animals particularly in the case of symptomatic dogs. The IgG subclass reactivity analysis revealed a broad specific recognition range of parasite recombinant antigens. Interestingly, differential profiles of IgG1 and IgG2 antibody reactivity were observed in asymptomatic and symptomatic dogs. The LmSIR2 protein was found to be a highly reactive molecule with IgG2 from most of the asymptomatic and symptomatic animals. Considering the fact that LmSIR2 secreted by the parasites can be bound and taken up by neighbouring cells, the latter could be a target for anti-LmSIR2 antibodies and this may contribute to the immunopathological alterations and host tissue damage. The implications of these observations in the pathogenesis of Leishmaniasis are discussed. PMID- 12644319 TI - Cloning and identification of a novel ubiquitin-like protein, BMSC-UbP, from human bone marrow stromal cells. AB - Ubiquitin is one of phylogenetically well-conserved proteins in all eukaryotes. Ubiquitin-dependent modification of protein contributes to fine regulation of cellular biological processes. Using large-scale screening of human bone marrow stromal cell (BMSC) cDNA library, we isolated a full-length cDNA of 1352 bp encoding 380 amino acids with a ubiquitin domain (UBQ), which was designed as bone marrow stromal cell-derived ubiquitin-like protein (BMSC-UbP). In addition to UBQ domain at its N-terminus, BMSC-UbP also possesses a ubiquitin-associated domain at its C-terminus, sharing moderate homology to some ubiquitin-like proteins such as UBIN, Chap1, and ubiquilin. BMSC-UbP localizes at chromosome 15q22.3-q23 as confirmed by blast search in human genome. BMSC-UbP mRNA is widely expressed in human multiple tissues and various tumor cell lines. Moreover, BMSC UbP mRNA decreased in BMSC stimulated with PMA and increased in HL60 cells stimulated with LPS, suggesting that BMSC-UbP might play roles in regulation of BMSC function or cell differentiation through an evocator- and cell-specific pattern. PMID- 12644320 TI - Advanced age negatively influences mesenteric lymph node T cell responses after burn injury. AB - While the pathophysiology of burn injury is well established in young adults, the factors that contribute to pathogenesis and increased death in elderly burn patients are not defined. The purpose of this study is to determine the effects of burn injury on mesenteric lymph node (MLN) T cell responses in young and aged mice. MLN is a cluster of lymph nodes that drains various parts of the intestine and is known to play role in clearance bacteria originating from the intestinal lumen. Results presented here suggest a significant suppression in Con A-induced MLN cell proliferation and IL-2 production in uninjured aged mice compared with uninjured young mice. Following 24 h after injury, although, a significant decrease in lymph node cell proliferation and IL-2 production was observed in both young and aged mice compared with their respective sham-injured animals, the suppression was more in aged mice. In addition we found a reduction in IFN-gamma, a Th-1 cytokine by MLN T cells from aged burned mice relative to young burn (P<0.05) or sham-injured mice (P<0.01). The Th-2 cytokine IL-4, on the other hand, was significantly increased in both young and aged burn-injured mice MLN T cells compared with their respective sham-injured mice. These results show that burn injury causes a greater suppression in MLN T cells ability to proliferate and a more pronounced shift to Th-2 phenotype in aged mice as compared with young mice. Such decreases in T cell functions may impair MLN's ability to clear the bacterial pathogens originating from intestine and thereby contribute to increased pathogenesis in injured host. PMID- 12644321 TI - Expression of PPARgamma in eosinophils and its functional role in survival and chemotaxis. AB - Eosinophils play a pivotal role in the mechanism of allergic diseases including asthma. Interleukin-5 (IL-5) and eotaxin are critical cytokines/chemokines for eosinophil activation. Peroxisome proliferator-activated receptor gamma (PPARgamma) is a nuclear receptor that regulates lipid metabolism. Recent evidence has suggested that PPARgamma serves as a negative regulator in the immune system. In the present study, we investigated the expression of PPARgamma and effect of PPARgamma agonist on human eosinophils. We demonstrated that purified eosinophils and Eol-1 cells express PPARgamma at the mRNA and protein levels. The PPARgamma agonist troglitazone reduced the IL-5-stimulated, but not spontaneous, eosinophil survival in a concentration-dependent manner. Moreover, the eotaxin-directed eosinophil chemotaxis was dose-dependently inhibited by troglitazone. Our results suggest that the administration of the PPARgamma agonists thiazolidinediones could be a new therapeutic modality for the treatment of allergic diseases such as asthma. PMID- 12644322 TI - Kinetic profiles of sequential gene expressions for chemokines in mice with contact hypersensitivity. AB - Using cDNA microarray technology, the expression of chemokine genes in the elicitation site of 2,4,6-trinitrochlorobenzene-induced contact hypersensitivity (CHS) was examined in mice. Of the 33 genes analyzed, levels of 11 gene expressions changed, and these can be assigned to four groups based on their kinetic patterns; (1) LARC/CCL20 whose mRNA level increased rapidly at 3 h post challenge and then gradually decreased, (2) JE/CCL2, MARC/CCL7, MIP-1gamma/CCL9, monocyte chemoattractant protein (MCP)-5/CCL12, ELC/CCL19 and BRAK/CXCL14 whose mRNA levels increased with time and reached the maximum at 6-9 h post-challenge, (3) LIX/CXCL5, Mig/CXCL9 and IP-10/CXCL10 whose mRNA levels increased gradually at least up to 12 h post challenge, and (4) SLC/CCL21 whose mRNA level decreased gradually with time after challenge. The findings suggest that sequential expression of chemokine genes is essential for orientating non-specific skin response to hapten-specific CHS response through the recruitment of inflammatory cells such as neutrophils, monocytes/macrophages and T-cells from the circulation into the tissue site. PMID- 12644323 TI - Elevated serum levels of thioredoxin in patients with acute exacerbation of asthma. AB - The pathogenesis of bronchial asthma is chronic airway inflammation caused by immune cells such as T lymphocytes and eosinophils. Eosinophils release cytotoxic products including reactive oxygen species at the site of inflammation, leading to epithelial damage. Human thioredoxin (TRX), a redox-regulating protein with antioxidant activity, is induced and secreted from cells by oxidative stress. This study was undertaken to investigate the clinical significance of TRX in the pathogenesis of asthma. We collected blood samples from 48 patients with bronchial asthma with or without attack, and measured serum ECP and pulmonary function as well as serum TRX. The serum TRX levels in patients with asthma were significantly increased in patients with mild (34.63 [28.40-42.73] ng/ml, medians with 25 and 75% interquartiles, P=0.0064) and moderate (38.83 [35.14-50.80] ng/ml, P=0.0017) asthma attacks compared with those during the asymptomatic period. The serum TRX levels were inversely correlated with FEV(1.0)% (r=-0.44, P=0.039) and %PEF (r=-0.49, P=0.020) during attack. There was a significant correlation between the serum TRX and the serum eosinophil cationic protein (rs=0.32, P=0.016). These findings suggest that serum TRX is related to the state of asthma exacerbation and allergic inflammation. PMID- 12644324 TI - Chronic murine Chagas' disease: the impact of host and parasite genotypes. AB - Chagas' disease is a protozoan infection caused by the flagellate Trypanosoma cruzi. Herein we utilise experimental infections of different mouse and parasite strains to investigate the relative importance of the host and parasite genotype, respectively, in causing Chagas' disease in mice. CBA/J and BALB/c mice infected with the Tulahuen strain of T. cruzi develop a severe acute disease characterised by transient parasitaemia and a high rate of mortality. While the acute phases in these mice are indistinguishable, they display differential outcomes of the infection since CBA/J mice eventually develop polymyositis and mild myocarditis whereas BALB/c mice are resistant to chronic disease. In contrast, BALB/c mice infected with the CA-1 clone of T. cruzi exhibit a mild acute phase, develop no polymyositis but do develop severe myocarditis. Thus both the parasite and host genotype, but not the severity of the acute phase, are important in determining the eventual outcome of T. cruzi infection. We also present a murine model suitable for investigating which host factors may be necessary to induce a chronic inflammatory disease after T. cruzi infection. PMID- 12644325 TI - Financial incentives, competition and a two tier service: lessons from the UK National Health Service internal market. AB - The experience of General medical practitioner (GP) fundholding is analysed for evidence of the response by family doctors to financial incentives. An analysis of consultant outreach, the local provision of out-patient services, in Scotland finds little evidence of a response, based on comparing the experience of fundholders with non-fundholders. At least in the case of hospital based services, financial incentives seem to be of secondary importance. Financial incentives for such services depend on consultant compliance which, arguably, was missing when it came to creating a two-tier service which deliberately favoured patients from fundholding practices. PMID- 12644326 TI - AIDS and public policy: the lessons and challenges of "success" in Thailand. AB - Thailand's public policy on AIDS is widely cited as one of the few examples of an effective national AIDS prevention program anywhere in the world. The Thai experience shows that a national response that mobilized key government and NGO partners and targeted the highest-risk transmission can be effective in reducing the scope of the epidemic, even when action is delayed. Based on interviews with policymakers, AIDS program managers, technical specialists, donors, and NGOs and on a review of the data, we highlight the lessons from public policy on AIDS in Thailand for other developing countries, review the state of the Thai epidemic and public policy in 2000, and identify three strategic priorities for the next phase of the response. PMID- 12644327 TI - Normative models of health technology assessment and the social production of evidence about telehealth care. AB - Telehealthcare is a rapidly growing field of clinical activity and technical development. These new technologies have caught the attention of clinicians and policy makers because they seem to offer more rapid access to specialist care, and the potential to solve structural problems around inequalities of service provision and distribution. However, as a field of clinical practice, telehealthcare has consistently been criticised because of the poor quality of the clinical and technical evidence that its proponents have marshalled. The problem of "evidence" is not a local one. In this paper, we undertake two tasks: first, we critically contrast the normative expectations of the wider field of Health Technology Assessment (HTA) with those configured within debates about Telehealthcare Evaluation; and second, we critically review models that provide structures within which the production of evidence about telehealthcare can take place. Our analysis focuses on the political projects configured within a literature aimed at stabilising evaluative knowledge production about telehealthcare in the face of substantial political and methodological problems. PMID- 12644329 TI - E-business in health care: does it contribute to strengthen consumer interest? AB - One of the goals of the reforms in the European health-care systems over the last two decades has been to make the health-care system more demand-oriented. There is not much known about the possible impact of E-business like approaches on this goal. This paper describes the concept of E-business. Two cases are introduced to illustrate the use of a simple E-business approach in a health-care setting. On the basis of these case studies, we aspect a reduction of the information disadvantages of patients. In our analysis, we also apply new institutional economy concepts, namely agency theory and transaction costs economics to focus on the position of the patient. Concluded is that it is more probable that preferences of demanders are answered by the suppliers of health care. PMID- 12644328 TI - Patient satisfaction with care is associated with personal choice of physician. AB - OBJECTIVE: To evaluate whether choosing one's own primary care doctor is associated with patient satisfaction with primary health care. To evaluate factors related to population's satisfaction with primary health care. POPULATION: A random sample of Estonian adult population (N=997). STUDY DESIGN: Cross-sectional study using a pre-categorized questionnaire which was compiled by the research group of the University of Tartu and the research provider EMOR. RESULTS: Altogether 68% of the respondents had been listed in their personal physician. Their overall satisfaction with the physician as well as satisfaction with several aspects of primary health care were significantly higher compared with those of unregistered respondents. Although some other factors (practice size, patient age, health status) also influenced patient satisfaction, presence of a personal physician appeared the most important predictor of high satisfaction with physician's punctuality and understanding, effectiveness of prescribed therapy, clarity of explanations given by the physician as well as with overall satisfaction with the physician. CONCLUSION: Personal doctor system is associated with patient satisfaction with different aspects of care. PMID- 12644330 TI - Stimulating pharmaceutical research and development for neglected diseases. AB - New vaccines and drug treatments are needed for tackling the neglected diseases (NDs) of poor countries. These diseases are associated with high levels of mortality and/or morbidity, but lack appropriate vaccines and drug treatments because of bacterial resistance, toxicity, long-treatment protocols, problems with administration or because none are available. Current initiatives directed at research and development (R&D) of NDs are being led predominantly by the governmental, inter-governmental and private not-for-profit sectors implemented by way of public-private partnerships. Push and pull mechanisms targeted at the pharmaceutical industry have also been proposed as another way to stimulate R&D of NDs; however, these should be viewed with some caution. Stimulating R&D for NDs is important as part of a wider long-term public health strategy and must be addressed simultaneously with resolving more immediate problems of access to medicines and health system sustainability in less developed countries. PMID- 12644331 TI - Cost-effectiveness of glycemic control and ophthalmological care in diabetic retinopathy. AB - AIMS: Glycemic control and ophthalmological care are known to significantly diminish the risk of visual impairment and blindness by diabetic retinopathy (DRP). The (cost-)effectiveness of both strategies was studied to highlight their benefits for patients and care providers. METHODS: A computer analysis was developed, following the progression of DRP and the effectiveness of metabolic control and ophthalmological care continuously and individually in cohorts of type I and type II DM patients with divergent degrees of compliance. Costs relate to present medical charges in the Netherlands. RESULTS: Intensive glycemic control shortens the duration of blindness in a type I DM patient by 0.76 years, intensive ophthalmological care by 0.53 years. One year sight gain may cost 1126 euros by providing ophthalmological care and 50479 euros by glycemic control. The duration of blindness drops in a type II DM patient by 0.48 and 0.13 years, respectively, whereas the effectiveness decreases as the age of onset of DM rises. CONCLUSIONS: The vast majority of diabetic patients benefits from both intensive glycemic control and intensive ophthalmological care, but these cost effective interventions which are not only complementary, but also substitute each other, require lasting, full compliance by all parties concerned. PMID- 12644332 TI - National Health Insurance and the antenatal care use: a case in Taiwan. AB - Many studies have found evidence for the importance of antenatal care on pregnancy outcomes. This paper focuses on the determinants of antenatal care use in Taiwan and provides a comparison of access to care before and after National Health Insurance (NHI) was implemented in 1995. A negative binomial model is applied to data from the 1989 and 1996 Taiwan Maternal and Infant Health Surveys to analyze antenatal care use. The results show that women in some situations had more antenatal care visits than average regardless of NHI implementation. These situations include: having a highly educated husband; gaining more weight than average during pregnancy; experiencing a first pregnancy; carrying twins or triplets; having care provided by a doctor rather than other caregivers; and switching to another health care facility during pregnancy. Regarding societal change, the trend toward delaying pregnancy is causing a change in care use. Additionally, three changes in care patterns after NHI are noteworthy. First, antenatal care visits at maternity clinics increased more than visits at hospitals. Second, before NHI's implementation, women who did blue-collar work or farm work sought care more frequently than housewives, but after NHI began government employees and businesswomen sought care more frequently. Third, antenatal care visits of mothers living in Taiwan's central area increased more than visits of those in the northern area. The expansion of medical care in aboriginal areas and outlying islands may prove to be one of NHI's best achievements. PMID- 12644333 TI - Health sector reforms in Kenya: an examination of district level planning. AB - The paper examines health sector reforms in Kenya at the district level based on the Government of Kenya's Health Policy Framework of 1994. The authors present the context of and historical perspective to health sector reforms in Kenya and discuss the major reform policies including decentralization to the district level. The authors then review intended policy outcomes, investigating assumptions on which the implementation and effectiveness of the reform agenda at the local level are based. The authors argue that emphasis on outcomes rather than process has not supported sustainable reforms or achieved the government's goal of improving health and ensuring equity for the citizens of the country. PMID- 12644334 TI - Introduction to "Optimizing management of non-ST-segment elevation acute coronary syndromes". Harmonizing advances in mechanical and pharmacologic intervention. PMID- 12644335 TI - Pathogenetic concepts of acute coronary syndromes. AB - The propensity of plaque to disrupt is a major determinant of future ischemic events. Although they are distinct from one another, the atherosclerotic and thrombotic processes appear to be interdependent and may be integrated under the term "atherothrombosis." It is now clear that plaque composition, rather than the percent stenosis, is a major determinant of plaque vulnerability. Plaque disruption seems to depend on both passive and active phenomena and is not purely mechanical. Inflammation (activation of monocytes/macrophages) is a major determinant of both plaque vulnerability and thrombogenicity as they relate to plaque disruption. In one-third of acute coronary syndromes, there is, however, no plaque disruption but only superficial erosion of a markedly stenotic, fibrotic plaque. In these cases, thrombus formation may be exacerbated by a hyperthrombogenic state present in patients with certain systemic risk factors. The endothelium plays a pivotal role in vascular homeostasis and hemostasis. This dynamic organ regulates blood thrombogenicity as well as contractile, secretory, and mitogenic activities in the vessel wall. Some classic risk factors induce endothelial dysfunction by reducing the bioavailability of nitric oxide, increasing tissue endothelin-1, and activating pro-inflammatory signaling pathways. Vascular hemostasis, which is the maintenance of blood fluidity and vascular integrity, is achieved by counter-balancing the intrinsic clotting tendency of blood. As a consequence of the central role of endothelial cells in hemostatic control, a dysfunctional endothelium will generate a pro-thrombotic environment favoring development of atherosclerotic lesions and thrombotic complications. PMID- 12644336 TI - Mechanisms of plaque vulnerability and rupture. AB - Rupture of atherosclerotic plaque has been identified as the proximate event in the majority of cases of acute ischemic syndromes. Plaque rupture exposes thrombogenic components of the plaque, activating the clotting cascade and promoting thrombus formation. Future culprit lesions are difficult to identify, however, and angiographic assessment of stenosis severity is prone to underestimation. Compared with plaques that cause severe luminal stenosis, vulnerable plaques may cause relatively minor stenosis, although they account for more cases of rupture and thrombosis. Such unstable, vulnerable plaques may be associated with outward remodeling of the vessel. Because severely stenotic plaques are more likely to stimulate collateral circulation to the post-stenotic segment, plaque rupture and thrombosis at such sites may be clinically silent. Characteristic histomorphologic features of vulnerable plaques include a high lipid content, increased numbers of inflammatory cells, and extensive adventitial and intimal neovascularity. The fibrous cap of an atherosclerotic plaque may become thin and rupture as a result of the depletion of matrix components through the activation of enzymes, such as matrix-degrading proteinases and cystine and aspartate proteases, and through the reduction in the number of smooth muscle cells. Activated T cells may also inhibit matrix synthesis through the production of interferon-gamma. A number of triggers of plaque rupture have been identified. Also, some thrombi may occur without rupture of the fibrous cap. Reducing the lipid component and inflammation in atherosclerotic plaques may help reduce the risk of plaque rupture. This may account for the clinical benefit of risk-factor reduction gained from changes in lifestyle and from drug therapy. PMID- 12644337 TI - Pharmacologic plaque passivation for the reduction of recurrent cardiac events in acute coronary syndromes. AB - Acute coronary syndrome (ACS) is often associated with the rupture of vulnerable atherosclerotic plaque, coronary thrombus formation, and abrupt limitation of blood flow, leading to adverse outcomes. Passivation of vulnerable plaque represents a therapeutic concept that has the potential to prevent or limit the magnitude of a new rupture in order to reduce the recurrence or severity of events. Plaque passivation can be defined as a process by which the structure or content of the atherosclerotic plaque is changed to reduce the risk of subsequent rupture and thrombosis. This may be achieved by using strategies that address different components of the plaque or the endothelium. The following factors can affect the susceptibility of plaque to rupture: macrophage infiltration; accumulation of inflammatory cells; paracrine secretion of enzymes that may cause degradation of the fibrous cap of coronary plaque; shear stress; circadian rhythm variation in stress hormone release; and infectious agents. The use of pharmacologic agents to reduce plaque vulnerability by passivation has been explored. Clinical studies demonstrate that lipid-modifying agents (e.g., statins), antiplatelet agents (acetylsalicylic acid, thienopyridines, thianopyridines, glycoprotein IIb/IIIa inhibitors), and antithrombotic agents (unfractionated heparin and low-molecular-weight heparin) can reduce the occurrence of acute coronary events in ACS patients. In addition, angiographic studies suggest that statins may also promote regression of atherosclerosis. Angiotensin-converting enzyme inhibitors, niacin, and calcium antagonists may also contribute to plaque passivation. This article reviews atherosclerotic plaque development and vulnerability and discusses some clinical studies highlighting the role of plaque passivation in the management of ACS patients. PMID- 12644338 TI - Troponin: an important prognostic marker and risk-stratification tool in non-ST segment elevation acute coronary syndromes. AB - Over the past decade, there has been a progressive evolution of cardiac marker testing in patients with acute coronary syndromes (ACS). This has not only resulted in a dramatic shift in how we view the diagnosis of myocardial infarction (MI), but it has also extended the role of cardiac marker testing into risk stratification and guidance of treatment decisions. By the year 2000, the development of highly sensitive and cardiac-specific troponin assays had resulted in a consensus change in the definition of MI, placing increased emphasis on cardiac-marker testing with troponins as the new gold standard. Furthermore, and perhaps more importantly, the role of the troponins as superior markers of subsequent cardiac risk in ACS patients became firmly established. Most recently, the supportive role of these markers in identifying patients with ACS who may derive particular benefit from potent anti-thrombotic and anti-platelet therapy or early invasive treatment strategies has been demonstrated. This paper will review the evolution of these important roles of troponin testing for risk stratification in ACS. PMID- 12644339 TI - C-reactive protein and other inflammatory risk markers in acute coronary syndromes. AB - Markers of myocyte necrosis such as cardiac troponin or creatine kinase myocardial band are invaluable tools for risk stratification among patients presenting with acute coronary syndromes (ACS). Nonetheless, many patients without any evidence of myocyte necrosis may be at high risk for recurrent ischemic events. In consideration of the important role that inflammatory processes play in determining plaque stability, recent work has focused on whether plasma markers of inflammation may help improve risk stratification. Of these markers, C-reactive protein (CRP) has been the most widely studied, and there is now robust evidence that CRP is a strong predictor of cardiovascular risk among apparently healthy individuals, patients undergoing elective revascularization procedures, and patients presenting with ACS. Moreover, even among patients with troponin-negative ACS, elevated levels of CRP are predictive of future risk. Other, more upstream markers of the inflammatory cascade, such as interleukin (IL)-6, have also been found to be predictive of recurrent vascular instability. A recent report from the second FRagmin during InStability in Coronary artery disease trial investigators suggests that elevated levels of an inflammatory marker such as IL-6 may indicate which patients may benefit most from an early invasive strategy. Other inflammatory markers currently under investigation include lipoprotein-associated phospholipase A(2), myeloperoxidase, and pregnancy-associated plasma protein A. Of all these novel markers, CRP appears to meet most of the criteria required for potential clinical application. Furthermore, the benefits of lifestyle modification and drug therapy with aspirin or statins may be most marked among those with elevated CRP levels. PMID- 12644341 TI - Glycoprotein IIb/IIIa inhibition in early intent-to-stent treatment of acute coronary syndromes: EPISTENT, ADMIRAL, CADILLAC, and TARGET. AB - The acute coronary syndromes (ACS), with or without ST-segment elevation, share a common pathophysiology of activated platelets and thrombin generation stimulated by plaque erosion and rupture. Both mechanical and pharmacologic treatment strategies have evolved in an attempt to improve reperfusion at the myocardial tissue level. Intracoronary stents have lowered the incidence of abrupt vessel closure and restenosis, while potent platelet inhibition from intravenous glycoprotein IIb/IIIa antagonists has reduced the rate of periprocedural myocardial infarction and late mortality. Abciximab has well-established clinical benefits in percutaneous revascularization trials, and several recent landmark studies have evaluated the efficacy of concomitant abciximab during mechanical reperfusion therapy in the setting of ACS. These trials are reviewed, and an overall perspective is provided. PMID- 12644340 TI - Small molecule glycoprotein IIb/IIIa receptor inhibitors as upstream therapy in acute coronary syndromes: insights from the TACTICS TIMI-18 trial. AB - Glycoprotein (GP) IIb/IIIa inhibitors are beneficial in unstable angina/non-ST segment elevation myocardial infarction (UA/NSTEMI). In large trials, the GP IIb/IIIa inhibitors tirofiban and eptifibatide were each found to reduce the risk of death or myocardial infarction (MI) in these patients at 30 days. These agents appear to be of greatest benefit in patients with a positive troponin at baseline, diabetes or ST-segment depression, recurrent angina, prior aspirin use, or a Thrombolysis In Myocardial Infarction (TIMI) risk score > or = 4. The Treat angina with Aggrastat and determine Cost of Therapy with an Invasive or Conservative Strategy (TACTICS) TIMI-18 trial was designed to compare the benefits of an early invasive versus a conservative strategy in high-risk UA/NSTEMI patients treated with GP IIb/IIIa inhibition. Patients were treated with tirofiban (for 48 h) plus aspirin and heparin and randomized to either invasive therapy (coronary angiography and revascularization when feasible) or conservative treatment (angiography only for patients with recurrent ischemia at rest or a positive stress test). A significant reduction in death or MI was demonstrated at 30 days (p = 0.02) and at 6 months (p = 0.0498). Death, MI, or rehospitalization for an acute coronary syndrome was also reduced with the invasive therapy at six months (p = 0.025). These results provide evidence to physicians that early GP IIb/IIIa inhibition in combination with a prompt invasive approach should be used more widely in UA/NSTEMI patients, particularly those at high risk. PMID- 12644342 TI - The role of low-molecular-weight heparin in the management of acute coronary syndromes. AB - A substantial number of clinical studies have consistently demonstrated that low molecular-weight heparin (LMWH) compounds are effective and safe alternative anticoagulants to unfractionated heparins (UFHs). They have been found to improve clinical outcomes in acute coronary syndromes and to provide a more predictable therapeutic response, longer and more stable anticoagulation, and a lower incidence of UFH-induced thrombocytopenia. Of the several LMWH agents that have been studied in large clinical trials, including enoxaparin, dalteparin, and nadroparin, not all have shown better efficacy than UFH. Enoxaparin is the only LMWH compound to have demonstrated sustained clinical and economic benefits in comparison with UFH in the management of unstable angina/ non-ST-segment elevation myocardial infarction (NSTEMI). Also, LMWH appears to be a reliable and effective antithrombotic treatment as adjunctive therapy in patients undergoing percutaneous coronary intervention. Clinical trials with enoxaparin indicate that LMWH is effective and safe in this indication, with or without the addition of a glycoprotein IIb/IIIa inhibitor. The efficacy demonstrated by enoxaparin in improving clinical outcomes in unstable angina/NSTEMI patients has led to investigations of its role in the management of ST-segment elevation myocardial infarction. Initial results are very encouraging, and they indicate that enoxaparin may potentially substitute for UFH as adjunctive therapy in fibrin specific thrombolytic regimens and improve coronary reperfusion rates in streptokinase-based regimens. PMID- 12644343 TI - Oral anticoagulants in patients with coronary artery disease. AB - Oral anticoagulants have been used in patients with vascular disease for over 40 years, yet their role in the secondary prevention of recurrent cardiovascular (CV) events remains controversial. The objectives of this systematic review are to more reliably determine the role of oral anticoagulants with and without antiplatelet therapy in patients with established coronary artery disease (CAD). Randomized trials in which oral anticoagulants were tested in CAD patients who were treated for at least three months were identified, and each trial was classified by the targeted level of intensity of anticoagulation. Data from the trials were combined using the modified Mantel-Haenszel method, and odds ratios were computed. Data from over 20,000 patients indicated that high-intensity oral anticoagulation (international normalized ratio [INR] >2.8) significantly reduced CV complications and increased bleeding compared with controls. Moderate intensity oral anticoagulation (INR 2 to 3) also reduced CV complications compared with controls. The combination of moderate-intensity oral anticoagulation and aspirin is more effective and equally as safe as aspirin alone. Low-intensity oral anticoagulation (INR <2) in the presence of aspirin does not reduce CV complications and increases bleeding compared with aspirin alone. PMID- 12644344 TI - The evolving role of direct thrombin inhibitors in acute coronary syndromes. AB - The central role of thrombin in the initiation and propagation of intravascular thrombus provides a strong rationale for direct thrombin inhibitors in acute coronary syndromes (ACS). Direct thrombin inhibitors are theoretically likely to be more effective than indirect thrombin inhibitors, such as unfractionated heparin or low-molecular-weight heparin, because the heparins block only circulating thrombin, whereas direct thrombin inhibitors block both circulating and clot-bound thrombin. Several initial phase 3 trials did not demonstrate a convincing benefit of direct thrombin inhibitors over unfractionated heparin. However, the Direct Thrombin Inhibitor Trialists' Collaboration meta-analysis confirms the superiority of direct thrombin inhibitors, particularly hirudin and bivalirudin, over unfractionated heparin for the prevention of death or myocardial infarction (MI) during treatment in patients with ACS, primarily due to a reduction in MI (odds ratio, 0.80; 95% confidence interval, 0.70 to 0.91) with little impact on death. The absolute risk reduction in the composite of death or MI at the end of treatment (0.8%) was similar at 30 days (0.7%), indicating no loss of benefit after cessation of therapy. Supportive evidence for the superiority of direct thrombin inhibitors over heparin derives from the recently reported Hirulog and Early Reperfusion or Occlusion (HERO)-2 randomized trial with ST-segment elevation ACS, which demonstrated a similar benefit of bivalirudin over heparin for the prevention of death or MI at 30 days (absolute risk reduction 1.0%), again primarily due to a reduction in MI during treatment (odds ratio, 0.70; 95% confidence interval, 0.56 to 0.87), with little impact on death. Further evaluation of hirudin and bivalirudin in the antithrombotic management of patients with ACS is warranted. PMID- 12644345 TI - Short- and long-term oral antiplatelet therapy in acute coronary syndromes and percutaneous coronary intervention. AB - Platelets play a central role in both the short- and long-term manifestations of atherothrombosis. In acute coronary syndrome (ACS), there is a steep rise in cardiovascular events early, followed by an incremental rise in cardiovascular events over the long term. This long-term event rate is related to persistent platelet activation and thrombin generation. There is therefore a need to optimize both short- and long-term oral antiplatelet and antithrombotic strategies. The benefits of aspirin therapy, when administered early and continued over the long term, were demonstrated in several early randomized trials. The Antithrombotic Trialists' Collaboration found a 46% reduction in vascular events with antiplatelet therapy (mostly aspirin). However, despite treatment with aspirin and proven therapies, recurrent events remain high. The adenosine diphosphate receptor antagonists, ticlopidine and clopidogrel, inhibit the early steps of platelet activation, degranulation, and release of prothrombotic and inflammatory mediators, while also preventing activation of the glycoprotein IIb/IIIa receptor. The Clopidogrel in Unstable angina to prevent Recurrent Events (CURE) trial demonstrated the benefits of aspirin plus clopidogrel in reducing major cardiovascular events (cardiovascular death, myocardial infarction [MI], and stroke reduced by 20%, p = 0.00009) in a broad range of patients with ACS when administered early and continued over the long term. The benefits emerge very rapidly after a 300 mg loading dose. For the large number of patients undergoing percutaneous coronary intervention in the CURE trial, there was a substantial risk reduction with clopidogrel pretreatment followed by long-term therapy (p < 0.002). This benefit was present, regardless of whether intervention was performed early or late. The significant benefits of aspirin and clopidogrel persist for the combined efficacy-safety end point of cardiovascular death, MI, stroke, or life-threatening bleeding when clopidogrel is started early, combined with aspirin and other standard therapies, and continued for up to one year. PMID- 12644347 TI - "Ischemia-guided" versus "early invasive" strategies in the management of acute coronary syndrome/non-ST-segment elevation myocardial infarction: the interventionalist's perspective. AB - Conventional therapy for non-ST-segment elevation acute coronary syndrome (ACS) has traditionally employed an "ischemia-guided" strategy. In this approach, diagnostic cardiac catheterization and revascularization are only used in patients with objective evidence of myocardial ischemia as identified by recurrent symptoms or provocative stress testing. More recent studies, however, have demonstrated improved clinical outcomes with the use of an "early invasive" approach, employing routine coronary angiography early in the patient's hospital course, followed by percutaneous intervention or bypass surgery where appropriate. Improved clinical outcomes associated with an "early invasive" strategy may have evolved as a consequence of recent advances in both adjunctive pharmacotherapy and revascularization technique. In particular, use of glycoprotein IIb/IIIa inhibitors and/or low-molecular-weight heparin before catheterization have been shown to reduce clinical events in patients with ACS, and may reduce the risk of an invasive approach by plaque passivation before interventional therapy. Perhaps more importantly, the combined use of glycoprotein IIb/IIIa inhibitors and intracoronary stenting may reduce the potential early hazard of an invasive approach by specifically decreasing the incidence of death and nonfatal myocardial infarction associated with percutaneous intervention. In spite of the benefits of this synergistic combination of pharmacology and mechanical revascularization, risk stratification remains important in identifying high-risk individuals most likely to benefit from an "early invasive" approach. In addition, angiography with possible percutaneous coronary intervention of "culprit" lesions should always be used in combination with aggressive medical therapy to treat the widespread coronary atherosclerosis commonly seen in patients with ACS. PMID- 12644346 TI - The thrombolysis in myocardial infarction risk score in unstable angina/non-ST segment elevation myocardial infarction. AB - Risk stratification in unstable angina (UA)/non-ST-segment elevation myocardial infarction (NSTEMI) can provide an estimate of a patient's prognosis and optimize clinical choices. The Thrombolysis In Myocardial Infarction (TIMI) risk score for UA/NSTEMI is an integrated approach that uses baseline variables that are part of the routine medical evaluation to identify patients at high risk for death and other major cardiac ischemic events. Using multivariable logistic regression, seven independent predictor variables were identified: age > or = 65 years, > or = 3 risk factors for coronary artery disease (CAD), known CAD (stenosis > or = 50%), severe anginal symptoms (> or = 2 anginal events in preceding 24 h), use of aspirin in the last seven days, ST-segment deviation > or = 0.05 mV, and elevated serum cardiac markers of necrosis. Each predictor carried similar prognostic weight; therefore, a risk score was constructed as the simple arithmetic sum of the number of predictors. The rate of death, MI, or urgent revascularization significantly increased as the TIMI risk score increased, ranging from < 5% for patients with a risk score of 0 or 1 to > 40% for patients with a risk score of 6 or 7. The risk score has been validated in several other trials of UA/NSTEMI. In addition, using the risk score to categorize patients also effectively defines a gradient for benefit with specific treatments such as low-molecular-weight heparins, glycoprotein IIb/IIIa inhibitors, and an early invasive strategy. PMID- 12644348 TI - Pathobiology, not angiography, should guide management in acute coronary syndrome/non-ST-segment elevation myocardial infarction: the non interventionist's perspective. AB - Although an early invasive strategy (angiography and percutaneous coronary intervention) is the convention in acute coronary syndrome (ACS)/non-ST-segment elevation myocardial infarction (MI) in the U.S., a conservative pharmacologic approach is common in other countries. Trial evidence has demonstrated a modest benefit with an angiographically guided approach; but patients having negative troponin values or who were receiving aspirin showed little or no benefit, and those without ST-segment changes had slightly worse outcomes. Limitations of angiography are clinically important. Identification of hemodynamically significant stenoses may be confounded by coronary remodeling. Also, most plaques, particularly those responsible for acute events, are extraluminal. Assessment of the luminal diameter of a lesion, which requires comparison with a normal reference segment, may be impossible because of the diffuse nature of the disease. Percutaneous coronary intervention after plaque rupture may itself cause embolization and no-reflow phenomena, leading to severe complications. In addition, most ruptures may be clinically silent. Evidence of a systemic inflammatory component suggests that ACS patients are at risk for plaque rupture at multiple sites. The inability of angiography to depict the true extent of atherosclerosis is supported by necropsy and transplant donor studies. A metabolic approach to this systemic disease is the only strategy designed to influence the behavior of both the small number of angiographically visible lesions and the large number of occult plaques. Statins and other agents decrease the incidence of death and MI by stabilizing atherosclerotic plaques throughout the coronary bed, reducing inflammation, collagen degradation, tissue factor expression, and vasomotor tone. PMID- 12644349 TI - "Routine invasive" versus "selective invasive" approaches to non-ST-segment elevation acute coronary syndromes management in the post-stent/platelet inhibition era. AB - Is a "routine invasive" or "selective invasive" strategy the best approach for patients with non-ST-segment elevation acute coronary syndrome (ACS)? A "selective invasive" strategy incorporates ischemia-guided use of aggressive medical therapy followed by angiography and revascularization for angina or stress-induced myocardial ischemia. The "routine invasive" strategy (cardiac catheterization followed by percutaneous coronary intervention within 24 to 48 h of symptom-onset) is frequently employed, but no randomized, controlled trials have demonstrated improved clinical outcomes. Recently, the second Fragmin and fast Revascularization during InStability in Coronary artery disease (FRISC-II) and the Treat angina with Aggrastat and determine Cost of Therapy with an Invasive or Conservative Strategy-Thrombolysis in Myocardial Infarction (TACTICS TIMI-18) trials found significant reductions in death, recurrent myocardial infarction, or hospitalization for biomarker-positive ACS. Also, the third Randomized Intervention Trial of unstable Angina (RITA-3) recently reported a halving of refractory angina and reduction in the use of antianginal medication with early intervention. Early trials failed to demonstrate the superiority of the "routine invasive" approach, presumably because of fewer revascularizations, unavailability of stents, and more recent use of glycoprotein IIb/IIIa inhibitors and low-molecular-weight heparins. The FRISC-II, TACTICS TIMI-18, and RITA-3 studies indicate that higher-risk patients benefit from early revascularization, but that aggressive antiplatelet, antithrombin, and anti-ischemic therapy are also important. While all three trials support an "early invasive" approach in intermediate- and high-risk patients, other trials support a more "conservative" approach in those without electrocardiographic changes or enzyme elevations. Optimal management should incorporate both strategies. PMID- 12644350 TI - A guide to therapeutic decision-making in patients with non-ST-segment elevation acute coronary syndromes. AB - Recent clinical trial evidence supports an inflammatory etiology in acute ischemic heart disease. When a segment of coronary artery becomes inflamed, important cytokines, such as tissue factor, are released, facilitating thrombosis. Serum inflammatory markers are elevated in most acute coronary syndrome patients at presentation. Mortality risk has been shown to be associated with increased levels of high-sensitivity C-reactive protein (CRP), interleukin 6, and serum vascular cell adhesion molecule. Platelets, which are rich in inflammatory mediators (CD40 and its ligand thrombospondin, and phospholipase A2), also supply important triggers for the inflammatory cascade. In addition, more than 35 platelet-associated messenger ribonucleic acid mediators involved in arterial injury and inflammation have been found. The use of biomarkers of inflammation, such as CRP, and of the sequelae of embolization, such as troponin, provide a window into the underlying pathophysiology of acute ischemic heart disease. New agents from three distinct drug classes have recently flooded the therapeutic armamentarium. Decision-making is further complicated by the choice of an invasive (aggressive) or a medical (conservative) strategy of management with respect to coronary revascularization. For patients at highest risk, aspirin, beta-blockers, nitrates, and a statin should be given, and clopidogrel, enoxaparin, a glycoprotein (GP) IIb/IIIa inhibitor, plus an invasive strategy should be considered. For intermediate- and low-risk patients, a "sliding-scale" approach may be best. Decisions about the three classes of antithrombotics--low molecular-weight heparins, GP IIb/IIIa inhibitors, and thienopyridines--along with whether to adopt an early invasive strategy, should be made on an individual basis. PMID- 12644351 TI - The relevance of differential response to trauma in an animal model of posttraumatic stress disorder. AB - BACKGROUND: Posttraumatic stress disorder affects 20%-30% of those exposed. Clinical studies employ stringent inclusion-exclusion criteria, yet animal studies include the entire exposed population as the study population. We examined the effect of grouping prestressed rats according to magnitude of response on the statistical analysis of results. METHOD: Response magnitude to predator exposure was assessed and used to group the animals into "diagnostic" groups. Two extremes were studied (clearly "maladapted" and clearly "well adapted" rats) using arbitrarily selected cutoff behavioral criteria (CBC). The data for the middle group were discarded for reasons of clarity. Hypothalamic pituitary-adrenal axis and heart-rate variability were analyzed for the entire exposed population and then according to the CBC. RESULTS: A single 10-min exposure to a predator caused fear-related behaviors in only 25.3% of exposed rats. Compared with control subjects and well-adapted exposed rats, maladapted rats exhibited significantly higher plasma corticosterone and corticotropin concentrations, increased sympathetic activity, diminished vagal tone, and increased sympathovagal balance. These differences surfaced only when data were analyzed according to CBC. Animals respond to stress heterogeneously, resembling humans. Overlooking this heterogeneity may obscure the results of data analysis. CONCLUSIONS: Animals can be divided into distinct groups according to magnitude of response and be studied accordingly. PMID- 12644352 TI - Cerebral function in posttraumatic stress disorder during verbal working memory updating: a positron emission tomography study. AB - BACKGROUND: This study examined cerebral function in posttraumatic stress disorder (PTSD) during the updating of working memory to trauma-neutral, verbal information. METHODS: Ten PTSD and matched control subjects completed a visuoverbal target detection task involving continuous updating (Variable target condition) or no updating (Fixed target condition) of target identity, with updating activity estimated by condition comparison. RESULTS: Normal updating activity using this paradigm involved bilateral activation of the dorsolateral prefrontal cortex (DLPFC) and inferior parietal lobe. The PTSD group lacked this activation in the left hemisphere and was significantly different from control subjects in this regard, but showed additional activation in the superior parietal lobe, bilaterally. CONCLUSIONS: The pattern of parietal activation suggests a dependence on visuospatial coding for working memory representation of trauma-neutral, verbal information. Group differences in the relative involvement of the DLPFC indicate less dependence in PTSD on the executive role normally attributed to the left DLPFC for monitoring and manipulation of working memory content in posterior regions of the brain. PMID- 12644353 TI - Neural correlates of anxiety associated with obsessive-compulsive symptom dimensions in normal volunteers. AB - BACKGROUND: The neural correlates of anxiety associated with obsessive-compulsive symptomlike provocation in normal volunteers are unknown. METHODS: Ten healthy volunteers participated in four functional magnetic resonance experiments. Subjects were scanned while viewing alternating blocks of emotional (normally aversive, washing-relevant, checking-relevant, or hoarding-relevant pictures) and neutral pictures, and imagining scenarios related to the content of each picture type. Nonparametric brain mapping analyses were used. RESULTS: In response to the provocative pictures in all experiments, increases in subjective anxiety and activation in bilateral ventral prefrontal, limbic, dorsal prefrontal, and visual regions were demonstrated. Anxiety related to different symptom dimensions was associated with different patterns of activation: provocation of washing-relevant anxiety predominantly activated dorsal and ventral prefrontal regions; checking relevant anxiety predominantly activated dorsal prefrontal regions; and hoarding relevant anxiety predominantly activated ventral prefrontal regions and the left amygdala. CONCLUSIONS: Our findings support a dimensional model of obsessive compulsive disorder (OCD) whereby 1) the brain systems implicated in the mediation of anxiety in response to symptom-related material in normal subjects are similar to those identified in OCD patients during symptom provocation, and 2) anxiety associated with different symptom dimensions is associated with differential patterns of activation of these neural systems. Further investigation of the neural basis of OCD symptom dimensions is required. PMID- 12644354 TI - Neocortical modulation of the amygdala response to fearful stimuli. AB - BACKGROUND: The cortical circuitry involved in conscious cognitive processes and the subcortical circuitry involved in fear responses have been extensively studied with neuroimaging, but their interactions remain largely unexplored. A recent functional magnetic resonance imaging (fMRI) study demonstrated that the engagement of the right prefrontal cortex during the cognitive evaluation of angry and fearful facial expressions is associated with an attenuation of the response of the amygdala to these same stimuli, providing evidence for a functional neural network for emotional regulation. METHODS: In the current study, we have explored the generalizability of this functional network by using threatening and fearful non-face stimuli derived from the International Affective Picture System (IAPS), as well as the influence of this network on peripheral autonomic responses. RESULTS: Similar to the earlier findings with facial expressions, blood oxygen level dependent fMRI revealed that whereas perceptual processing of IAPS stimuli was associated with a bilateral amygdala response, cognitive evaluation of these same stimuli was associated with attenuation of this amygdala response and a correlated increase in response of the right prefrontal cortex and the anterior cingulate cortex. Moreover, this pattern was reflected in changes in skin conductance. CONCLUSIONS: The current results further implicate the importance of neocortical regions, including the prefrontal and anterior cingulate cortices, in regulating emotional responses mediated by the amygdala through conscious evaluation and appraisal. PMID- 12644355 TI - Neural circuitry underlying voluntary suppression of sadness. AB - BACKGROUND: The ability to voluntarily self-regulate negative emotion is essential to a healthy psyche. Indeed, a chronic incapacity to suppress negative emotion might be a key factor in the genesis of depression and anxiety. Regarding the neural underpinnings of emotional self-regulation, a recent functional neuroimaging study carried out by our group has revealed that the dorsolateral prefrontal cortex (DLPFC) and anterior cingulate cortex are involved in voluntary suppression of sexual arousal. As few things are known, still, with respect to the neural substrate underlying volitional self-regulation of basic emotions, here we used functional magnetic resonance imaging to identify the neural circuitry associated with the voluntary suppression of sadness. METHODS: Twenty healthy female subjects were scanned during a Sad condition and a Suppression condition. In the Sad condition, subjects were instructed to react normally to sad film excerpts whereas, in the Suppression condition, they were asked to voluntarily suppress any emotional reaction in response to comparable stimuli. RESULTS: Transient sadness was associated with significant loci of activation in the anterior temporal pole and the midbrain, bilaterally, as well as in the left amygdala, left insula, and right ventrolateral prefrontal cortex (VLPFC) (Brodmann area [BA] 47). Correlational analyses carried out between self-report ratings of sadness and regional blood oxygen level dependent (BOLD) signal changes revealed the existence of positive correlations in the right VLPFC (BA 47), bilaterally, as well as in the left insula and the affective division of the left anterior cingulate gyrus (BA 24/32). In the Suppression condition, significant loci of activation were noted in the right DLPFC (BA 9) and the right orbitofrontal cortex (OFC) (BA 11), and positive correlations were found between the self-report ratings of sadness and BOLD signal changes in the right OFC (BA 11) and right DLPFC (BA 9). CONCLUSIONS: These results confirm the key role played by the DLPFC in emotional self-regulation. They also indicate that the right DLPFC and right OFC are components of a neural circuit implicated in voluntary suppression of sadness. PMID- 12644356 TI - Neuronal substrates of sensory gating within the human brain. AB - BACKGROUND: For the human brain, habituation to irrelevant sensory input is an important function whose failure is associated with behavioral disturbances. Sensory gating can be studied by recording the brain's electrical responses to repeated clicks: the P50 potential is normally reduced to the second of two paired clicks but not in schizophrenia patients. To identify its neural correlates, we recorded electrical traces of sensory gating directly from the human hippocampus and neocortex. METHODS: Intracranial evoked potentials were recorded using hippocampal depth electrodes and subdural strip and grid electrodes in 32 epilepsy patients undergoing invasive presurgical evaluation. RESULTS: We found evidence of sensory gating only in the hippocampus, the temporo parietal region (Brodmann's areas 22 and 2), and the prefrontal cortex (Brodmann's areas 6 and 24); however, whereas neocortical habituating responses to paired clicks were peaking around 50 msec, responses within the hippocampus proper had a latency of about 250 msec. CONCLUSIONS: Consistent with data from animal studies, our findings show that the hippocampus proper contributes to sensory gating, albeit during a time window following neocortical habituation processes. Thus, sensory gating may be a multistep process, with an early phase subserved by the temporo-parietal and prefrontal cortex and a later phase mediated by the hippocampus. PMID- 12644357 TI - Congenitally learned helpless rats show abnormalities in intracellular signaling. AB - BACKGROUND: Affective disorders and the drugs used to treat them lead to changes in intracellular signaling. We used a genetic animal model to investigate to what extent changes in intracellular signal transduction confer a vulnerability to mood or anxiety disorders. METHODS: Levels of gene expression in a selectively bred strain of rats with a high vulnerability to develop congenitally learned helplessness (cLH), a strain highly resistant to the same behavior (cNLH) and outbred Sprague-Dawley (SD) control animals were compared using quantitative reverse transcription polymerase chain reaction. RESULTS: Congenitally learned helpless animals had a 24%-30% reduced expression of the cyclic adenosine monophosphate response element binding protein messenger ribonucleic acid (mRNA) in the hippocampus and a 40%-41% increased level of the antiapoptotic protein bcl 2 mRNA in the prefrontal cortex compared to cNLH and SD rats. Other significant changes included changes in the expression levels of the alpha catalytic subunit of protein kinase A, glycogen synthase kinase 3beta, and protein kinase C epsilon. CONCLUSIONS: Congenitally learned helpless animals show evidence of altered signal transduction and regulation of apoptosis compared to cNLH and SD control animals. PMID- 12644358 TI - Regulation of GAP-43 expression by chronic desipramine treatment in rat cultured hippocampal cells. AB - BACKGROUND: The importance of molecular and cellular changes in hippocampus in major depression and in the mechanism of action of antidepressants has become increasingly clear. Identification of novel targets for antidepressants in hippocampus is important to understanding their therapeutic effects. METHODS: We used cDNA microarray to measure the expression patterns of multiple genes in primary cultured rat hippocampal cells. In situ hybridization and Northern and immunoblotting analysis were used to determine brain regional distribution and mRNA and protein levels of target genes. RESULTS: After comparing hybridized signals between control and desipramine treated groups, we found that chronic treatment with desipramine increased the expression of six genes and decreased the expression of two genes. One of the upregulated genes is growth associated protein GAP-43. In situ hybridization revealed that desipramine increased GAP-43 gene expression in dentate gyrus but not other brain regions. Northern and immunoblotting analysis revealed that desipramine increased GAP-43 mRNA and protein levels. GAP-43 expression is also increased by another antidepressant, tranylcypromine, but not by lithium or haloperidol. CONCLUSIONS: Because GAP-43 regulates growth of axons and modulates the formation of new connections, our findings suggest that desipramine may have an effect on neuronal plasticity in the central nervous system. PMID- 12644359 TI - Substance P serum levels are increased in major depression: preliminary results. AB - BACKGROUND: Substance P (SP) is thought to have an impact in the pathophysiology of depression and the mechanism of action of antidepressant drugs. The aim of this study was to analyze the serum SP levels in healthy control subjects and in depressed patients before and after antidepressant treatments. METHODS: Twenty three patients with major depression and 33 control subjects participated in the study. Using an enzyme immunoassay, the SP serum levels were determined in patients at baseline (before treatment) and after 2 and 4 weeks of antidepressant therapy. Determinations of SP in control subjects were carried out twice, at baseline and after 4 weeks. RESULTS: The mean baseline SP serum concentration was significantly higher in depressed patients as compared with control subjects (p <.001). Repeated measurements in control subjects showed that SP remains relatively constant over a period of 4 weeks. Although in depressed patients there was no overall change in the mean SP levels between baseline and 4 weeks' treatment, 37% of them exhibited a decrease of SP (15%-50%), which can be correlated to a better drug response than an increase in SP concentration after treatment (p =.001). CONCLUSIONS: Our data show that serum SP levels are increased in a proportion of patients with major depression and might thus indicate a subgroup of the disorder in which neuropeptides have a key position. Future studies are needed to clarify whether the observed SP decrease in treatment responders can be attributed to a specific class of drugs. PMID- 12644360 TI - Phenylalanine hydroxylase gene in psychiatric patients: screening and functional assay of mutations. AB - BACKGROUND: Reports relating phenylalanine kinetics and metabolism to psychiatric disorders led us to undertake the comprehensive screening of the phenylalanine hydroxylase (PAH) coding region and functional testing of discovered mutations in a sample of psychiatric patients and healthy control subjects. METHODS: Genomic DNA from psychiatric patients and control subjects was assayed for sequence variants in all PAH coding regions and splice junctions. In vivo functional analysis of mutations was conducted by assessing the kinetics and conversion to tyrosine of a standardized phenylalanine dose and by measuring fasting pterin levels. RESULTS: A known missense mutation was observed in a schizoaffective subject, and a novel missense mutation was discovered in four subjects with schizophrenia and one normal subject. The schizoaffective patient heterozygous for the known A403V mutation showed the lowest rate of phenylalanine kinetics and lowest conversion to tyrosine in the patient sample. The four schizophrenic patients heterozygous for the novel K274E mutation showed significantly decreased phenylalanine kinetics, reduced conversion to tyrosine, and increased synthesis of the PAH cofactor tetrahydrobiopterin compared with schizophrenic subjects without the mutation. CONCLUSIONS: The study findings suggest that larger scale studies are warranted to test the relationship of the PAH genotype with a psychiatric phenotype. PMID- 12644362 TI - Influence of micelle solubilization by tocopheryl polyethylene glycol succinate (TPGS) on solubility enhancement and percutaneous penetration of estradiol. AB - The effect of micellar solubilization on the enhancement of the solubility and percutaneous penetration of estradiol by the surface-active agent, tocopheryl polyethylene glycol succinate (TPGS) was characterized in this study. Results show that the solubility of estradiol was improved in the presence of TPGS through micellar solubilization. The critical micelle concentration (CMC) of TPGS increased with increasing ethanol concentration in the medium. With the flux corrected to the saturated level (J(corrected)) of the free form of estradiol, an increase in the alcohol content of the medium resulted in an increase in J(corrected) for all levels of TPGS examined. For the same level of alcohol content, an increase in the TPGS concentration mostly led to a small extent of decrease in J(corrected). However, the extent of decrease was more obvious in media containing more than 60% alcohol. We also confirmed that only an insignificant amount of TPGS was transported across the skin (below the detection limit of 2 microg/ml). Permeabilities (P(eff)), which describe the overall effects (DK/H) on the stratum corneum (SC), decreased with increasing TPGS concentration for media containing 0, 40, 60, and 80% alcohol, whereas they increased then decreased with increasing TPGS concentration for media containing 10 and 20% alcohol. The enhancement ratios based on P(eff) assuming that the medium contained 0% TPGS and alcohol as unity did not increase accordingly with increases in TPGS concentration at the same level as alcohol. Likewise, the enhancement ratios for the same level of TPGS increased with low alcohol content, but then decreased with increasing alcohol content. We concluded that micellar solubilization by TPGS was able to improve the solubility of estradiol, but it only had an insignificant influence on the skin. Interfacial coverage of TPGS with increasing TPGS concentration and hindrance of the partitioning of estradiol by the increasing alcohol content might play a role in influencing the permeability of estradiol. PMID- 12644363 TI - Reversible lipidization for the oral delivery of salmon calcitonin. AB - Salmon calcitonin (sCT), a 32-amino-acid polypeptide, was lipidized by using a reversible aqueous lipidization (REAL) technology. When injected subcutaneously into mice, the AUC of REAL-sCT was four times greater than that of sCT and a similar pattern of reduction in plasma calcium level was observed. The therapeutic effect of REAL-sCT was evaluated in ovariectomized (OVX) rats. The development of osteoporosis in OVX rats was determined by measuring the urinary level of deoxypyridinoline (DPD), a biochemical marker of bone resorption. It was found that the DPD levels were significantly reduced in rats that were orally administered a dose of 50 microg/kg/day of REAL-sCT. No reduction in urinary DPD levels could be detected in OVX rats treated similarly with unmodified sCT. In addition, significant levels of sCT were detected in rat plasma up to 12 h after oral administration of REAL-sCT at 500 microg/kg, while the plasma concentration of sCT was undetectable at 1 h after oral administration with the same dose of sCT. The AUC of oral REAL-sCT was at least 19 times higher than that of sCT. Our results indicate that reversibly lipidized polypeptides exhibit not only improved pharmacokinetic and pharmacodynamic behaviors, but also an enhanced oral bioavailability. PMID- 12644364 TI - Preparation and evaluation of a melt pelletised paracetamol/stearic acid sustained release delivery system. AB - The potential of a sustained release formulation for paracetamol produced by melt pelletisation was investigated. The chosen formulation was based on the combination of stearic acid as a melting binder and anhydrous lactose as a filler. After determination of the size distribution, the pellet characterisation included scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), specific surface area and true density determination. Hence, the in vitro release from every single size fraction (2000, 1250, 800, 630, <630 microm) was evaluated and the release mechanism was analysed with the help of an appropriate mathematical model. The results of drug content and superficial atomic composition were found to be constant in all pellets size fractions, attesting the ability of melt pelletisation in a high shear mixer to form a product with homogeneous composition. The mathematical model is built on the hypotheses that drug diffusion and solid drug dissolution in the release environment are the key phenomena affecting drug release kinetics. Smaller classes apart (particles are not perfectly spherical), the comparison between model best fitting and experimental data indicated the reasonability of these hypotheses. Moreover, model reliability is proved by its ability of predicting drug release from a known mixture of the above mentioned particles classes. PMID- 12644365 TI - Evaluation of the mucosal irritation potency of co-spray dried Amioca/poly(acrylic acid) and Amioca/Carbopol 974P mixtures. AB - The purpose of this study was to evaluate the biocompatibility of different Amioca/poly(acrylic acid) and Amioca/Carbopol 974P co-spray dried mixtures with an alternative mucosal irritation test using slugs. The irritation potential of the mixtures was measured by the amount of mucus produced during a repeated 30 min contact period. Additionally, membrane damage was assessed by measuring the protein and enzyme release from the body wall of slugs after treatment. All the Amioca/poly(acrylic acid) co-spray dried mixtures (50:50 and 25:75 ratios) induced slight irritation of the mucosal tissue as was demonstrated by the significantly increased mucus production however no increased protein and enzyme release was detected. Co-spray dried Amioca/Carbopol 974P mixtures containing 40% and more Carbopol 974P demonstrated a significantly higher mucus production and release of cytosolic LDH, indicating membrane damage. The total mucus production of the slugs treated with the co-spray dried mixtures containing up to 20% Carbopol 974P was significantly higher compared to the blank slugs. However, these mixtures induced no membrane damage since no additional effect on the protein release and no enzyme release was detected. By co-spray drying up to 20% Carbopol 974P could be incorporated without showing a distinct sign of irritation. These mixtures can be considered as potentially safe bioadhesive carriers. PMID- 12644366 TI - Optimization of a thermally reversible W/O/W multiple emulsion for shear-induced drug release. AB - PURPOSE: The present work aimed at improvement of the formulation of a previously developed thermo-reversible W/O/W multiple emulsion by increasing the emulsion stability and reaching a higher fraction of an encapsulated drug released under shear. The emulsion was based on high molecular weight graft-copolymers of poly(acrylic acid) and Pluronic F127 as stabilizing agents. METHODS: Once a stable W/O/W thermo-reversible multiple emulsion was obtained via a fine-tuning of the formulation, rheological, granulometric and conductometric tests were performed to assess the thermo-reversible behavior and the fragmentation-release characteristics of the new W/O/W multiple emulsion. RESULTS: The emulsion exhibited a 10(3) fold increase in viscosity over a range of temperatures from 20 to 40 degrees C. At moderate shearing, a complete release of the marker encapsulated in the internal aqueous phase was observed (99.6%) at 35 degrees C, whereas only 30% was released at 20 degrees C. Under similar conditions at 35 degrees C, slightly more than 50% was released for the initial formula. CONCLUSION: Additionally, the ease of fabrication of the thermo-reversible W/O/W multiple emulsion combined with the complete release under shear at body temperature and the superior emulsion stability suggest numerous applications in the controlled release of drugs. PMID- 12644367 TI - Non-degradable microparticles containing a hydrophilic and/or a lipophilic drug: preparation, characterization and drug release modeling. AB - Non-degradable microparticles based on ammonio methacrylate copolymers (Eudragit RS:RL 4:1 blends) containing the hydrophilic drug propranolol HCl and/or the lipophilic drug nifedipine were prepared with an oil-in-water (O/W) and a water in-oil-in-water (W/O/W) solvent evaporation technique. Both drugs were successfully incorporated separately as well as simultaneously. In all cases, the resulting release rate(s) of the drug(s) was/were found to be controlled over periods of at least 8 h. To elucidate the underlying mass transport mechanisms, the microparticles were thoroughly characterized by X-ray powder diffractometry, differential scanning calorimetry, particle size analysis, and determination of the actual drug loading(s). Analytical solutions of Fick's second law of diffusion considering non-steady state conditions were used to describe the release of propranolol HCl. Interestingly, the resistance for drug release within the unstirred liquid boundary layers on the surfaces of the microparticles was found to be negligible compared to the diffusional resistance within the polymeric devices. Importantly, the mathematical theories could be used to normalize the experimentally determined in vitro drug release with respect to the microparticle size. Thus, the effect of the type of preparation method (O/W vs. W/O/W) and device composition (polymer blend plus one drug only vs. polymer blend plus drug combination) on the diffusional resistance within the microparticles could be studied. In addition, further insight into the occurring mass transport processes was gained. For example, the time-dependent evolution of the drug concentration profiles within the microparticles upon exposure to the release medium could be calculated. An interesting practical application of the mathematical theories is the possibility to predict the effect of different formulation parameters on the resulting drug release patterns, e.g. the effect of the microparticle size. PMID- 12644368 TI - Anionic polyethyleneglycol lipids added to cationic lipoplexes increase their plasmatic circulation time. AB - Cationic liposomes have been widely sensed as good DNA compacting delivery agents. Although their use generally met with encouraging results in vitro, the results in vivo were rather disappointing, as they strongly interact with the blood components before they can reach the therapeutic target. Polyethyleneglycol (PEG) shielding has been proposed as a way to alleviate this effect, but was still found unsatisfactory in most instances for systemic administration. We demonstrate here that the insertion of anionic functions between the lipid part and the PEG, at a correct distance to favor electrostatic interactions with the outer cationic layer of the lipoplexes, provides not only a decrease in the mean peripheral charge of the lipoplex (zeta potential), but also a greater colloidal stability of the particles in the presence of serum. Transfection in the lung is also decreased with negatively charged PEG shielding, although no significant changes are observed in the tumor. This encouraging new approach should consequently be combined with active extra-cellular receptor targeting to achieve the desired delivery of the therapeutic DNA to tumor tissues. PMID- 12644370 TI - Synthesis of octyl arabinofuranosides as substrates for mycobacterial arabinosyltransferases. AB - A panel of octyl oligosaccharides comprised of arabinofuranose rings have been synthesized via efficient and readily scaleable routes. The key glycosylation reactions involved the coupling of octyl glycoside acceptors with the appropriate thioglycosides using N-iodosuccinimide and silver triflate activation. These syntheses were undertaken to provide substrates suitable for use in assays of mycobacterial arabinosyltransferases. PMID- 12644371 TI - Synthesis of the pentasaccharide related to the repeating unit of the antigen from Shigella dysenteriae type 4 in the form of its methyl ester 2 (trimethylsilyl)ethyl glycoside. AB - Starting from D-mannose, D-glucose and L-fucose, the pentasaccharide derivative methyl 2,3,4-tri-O-benzyl-alpha-L-fucopyranosyl-(1-->3)-2-O-acetyl-4,6-O benzylidene-alpha-D-mannopyranosyl-(1-->3)-2-O-acetyl-6-O-benzyl-4-O-(2,3,4-tri-O benzyl-alpha-L-fucopyranosyl)-alpha-D-mannopyranosyl-(1-->4)-[2 (trimethylsilyl)ethyl 2,3-di-O-benzyl-beta-D-glucopyranosid]uronate was synthesized. This compound with two alpha-mannopyranosyl units was transformed, via Walden inversion and subsequent deprotection, into the alpha-D-glucosamine type target compound, namely methyl alpha-L-fucopyranosyl-(1-->3)-2-acetamido-2 deoxy-alpha-D-glucopyranosyl-(1-->3)-2-acetamido-2-deoxy-4-O-(alpha-L fucopyranosyl)-alpha-D-glucopyranosyl-(1-->4)-[2-(trimethylsilyl)ethyl beta-D glucopyranosid]uronate which is related to the repeating unit of the O-antigen from Shigella dysenteriae type 4. PMID- 12644372 TI - Characterization of an acetylated heteroxylan from Eucalyptus globulus Labill. AB - A heteroxylan was isolated from Eucalyptus globulus wood by extraction of peracetic acid delignified holocellulose with dimethyl sulfoxide. Besides (1-->4) linked beta-D-xylopyranosyl units of the backbone and short side chains of terminal (1-->2)-linked 4-O-methyl-alpha-D-glucuronosyl residues (MeGlcA) in a 1:10 molar ratio, this hemicellulose contained galactosyl and glucosyl units attached at O-2 of MeGlcA originating from rhamnoarabinogalactan and glucan backbones, respectively. About 30% of MeGlcA units were branched at O-2. The O acetyl-(4-O-methylglucurono)xylan showed an acetylation degree of 0.61, as determined by 1H NMR spectroscopy, and a weight-average molecular weight (M(w)) of about 36 kDa (P=1.05) as revealed from size-exclusion chromatography (SEC) analysis. About half of the beta-D-xylopyranosyl units of the backbone were found as acetylated moieties at O-3 (34 mol%), O-2 (15 mol%) or O-2,3 (6 mol%). Practically, all beta-D-xylopyranosyl units linked at O-2 with MeGlcA residues were 3-O-acetylated (10 mol%). PMID- 12644373 TI - Structural analysis of the Lactobacillus rhamnosus strain KL37C exopolysaccharide. AB - The exopolysaccharide from the lactic acid bacterium Lactobacillus rhamnosus strain KL37C isolated from human intestinal flora was prepared by sonication of bacterial cell mass suspended in water followed by centrifugation and cold ethanol precipitation of the supernatant. The polysaccharide material was purified by gel permeation chromatography on an TSK HW-50 column and characterised using chemical and enzymatic methods. On the basis of sugar and methylation analysis and 1H, 13C, 1D and 2D NMR spectroscopy the exopolysaccharide was shown to be composed of the following pentasaccharide repeating unit:-->3)-alpha-D-Glcp-(1-->2)-beta-D-Galf-(1-->6)-alpha-D-Galp-(1- >6)-alpha-D-Glcp-(1-->3)-beta-D-Galf-(1--> PMID- 12644374 TI - Molecular fractionation of starch by density-gradient ultracentrifugation. AB - Amylose and amylopectin in corn and potato starches were fractionated by centrifugation at 124,000g for 3-72 h at 40 degrees C in a gradient media, Nycodenz, based on their sedimentation rate differences. The fractions were collected from a centrifuge tube, and then analyzed by the phenol-sulfuric acid method and iodine-binding test. Amylopectin, a large and highly branched starch molecule, migrated faster than amylose and quickly reached its isopycnic point with a buoyant density of about 1.25 g/mL, exhibiting a sharp and stable carbohydrate peak. Amylose, which is a relatively small and linear molecule, however, migrated slowly in a broad density range and continued moving to higher density regions, eventually overlapping with amylopectin peak as the centrifugation continued. This could indicate that the buoyant density of amylose is similar to that of amylopectin. Under centrifugal conditions of 3 h and 124,000g, amylose and amylopectin molecules were clearly separated, and the presence of intermediate starch molecules (11.5 and 7.7% for corn and potato starch, respectively) was also observed between amylose and amylopectin fractions. The amylose content of corn and potato starches was 22.6 and 21.1%, respectively, based on the total carbohydrate analysis after the ultracentrifugation for 3 h. In alkaline gradients (pH 11 or 12.5), the sedimentation rate of starch molecules and the buoyant density of amylopectin were reduced, possibly due to the structural changes induced by alkali. PMID- 12644375 TI - Structural features of an arabinogalactan gum exudates from Spondias dulsis (Anacardiaceae). AB - The tree Spondias dulcis, located in Venezuela, exudes a light-brown gum. The polysaccharide, isolated from the original gum, contains galactose, arabinose, mannose, rhamnose, glucuronic acid, and its 4-O-methyl derivative. Application of chemical methods, in combination with 1D and 2D NMR spectroscopy afforded interesting structural features of the gum polysaccharide. The unequivocal presence of rhamnose in the polymer structure was confirmed by chemical and spectral data [1H (1.03 ppm); 13C (16.92 ppm)]. Also confirmed was the existence of 3-O- and 6-O-substitutes galactose residues by the spectral data correlations observed in Heteronuclear Multiple Quantum Coherence (HMQC) and Heteronuclear Multiple Bond Correlation (HMBC). Also observed were unequivocal resonances for beta-D-glucuronic acid and its 4-O-methyl derivative, and the presence of 3-O alpha-L-arabinofuranose and 3-O-beta-L-arabinopyranose residues. PMID- 12644376 TI - Macrocycle conformation and self-inclusion phenomena in octakis(3-O-butanoyl-2,6 di-O-pentyl)-gamma-cyclodextrin (Lipodex E) by NMR spectroscopy and molecular dynamics. AB - Octakis(3-O-butanoyl-2,6-di-O-pentyl)-gamma-cyclodextrin (Lipodex E) is a lipophilic chiral selector successfully used for the enantioselective gas chromatographic separation of a multitude of racemic analytes. NMR data (13C chemical shifts, 3J(HH), rotating frame NOEs (ROEs)) and molecular dynamics (MD) simulations point out that the macrocycle is distorted with respect to the canonical truncated-cone shape of native cyclodextrins, although C(8) symmetry is retained on the NMR timescale. ROE data and MD trajectories provide evidence for self-inclusion of one 6-O-pentyl pendant chain within the cavity of Lipodex E. The interpretation of long-range and low-intensity ROEs is supported by the calculation of average internuclear distances by using the radial distribution function (RDF) calculated from MD trajectories. MD simulations are eventually used to compare the flexibility of the macrocycle of Lipodex E with that of native gammaCD. PMID- 12644378 TI - The three-dimensional structure of the mega-oligosaccharide rhamnogalacturonan II monomer: a combined molecular modeling and NMR investigation. AB - In this study, we describe the first optimized molecular models of the mega oligosaccharide rhamnogalaturonan II, that is found in the primary cell walls of all higher plants. The 750 MHz 1H NMR data previously reported and new heteronuclear correlation spectra (sensitivity-enhanced HSQC and HSQC-TOCSY) were first reassigned in light of the modifications in the primary structure. In turn, the experimental NMR data revealed the presence of an additional sugar, alpha Araf (E-chain), and also the disaccharidic repeating unit of RG-I, another component of the pectic matrix. Due to a fuller picture of the primary structure of RG-II, a much more complete assignment of the NOE data has been achieved. A systematic computational study based on these NOEs lead us to a realistic three dimensional description of the RG-II, in excellent agreement with the molecular dimensions obtained from various experimental methods. PMID- 12644377 TI - Chemically methylated and reduced pectins: preparation, characterisation by 1H NMR spectroscopy, enzymatic degradation, and gelling properties. AB - The gelling properties of pectins are known to be closely related to the degree of methylation (DM) and the distribution of the ester groups. In order to investigate this dependency, a natural citrus pectin (DM 64%) has been methylated to pectins with higher DM or saponified to achieve pectins with lower DM. A simple method for determination of DM by 1H NMR spectroscopy is presented. New modified pectins have been prepared by treatment of pectins having different DM with NaBH(4) to reduce selectively the methyl esters to primary alcohols in the presence of free acids. The degree of reduction (DR) and the DM of the remaining carboxylic acids could likewise be determined by 1H NMR spectroscopy. The new reduced pectins are recognized by the pectin degrading enzymes polygalacturonase PGI and PGII as well as by pectin lyase, all from Aspergillus niger, but the enzymes exhibit lower specific activities as compared with unmodified pectin. The new reduced pectins exhibit high gelling properties. PMID- 12644379 TI - Synthesis of new branched-chain amino sugars. AB - 1,3-Dipolar cycloaddition of methylideneaniline N-oxide to sugar enones is described. The addition occurred exclusively from the side opposite to the aglycone affording the corresponding alkyl alpha-D-lyxo-hexopyranosid-(2,3:5',4') phenylisoxazolidin-4-uloses. Hydrogenation of these compounds readily yielded the corresponding alkyl 3-deoxy-3-N-phenylaminomethyl-alpha-D-talopyranoside, that were readily transformed to the acetates. The structure and conformation of the bicyclic compounds were determined by 1H NMR studies and semi-empirical molecular orbital calculations employing the AM1 method. PMID- 12644380 TI - Efficient selective preparation of methyl-1,2,4-tri-O-acetyl-3-O-benzyl-beta-L idopyranuronate from methyl 3-O-benzyl-L-iduronate. AB - Methyl 1,2,4-tri-O-acetyl-3-O-benzyl-L-idopyranuronate 6beta/6alpha, prepared from methyl 3-O-benzyl-L-iduronate (4), is a key synthon in heparin/heparan sulfate synthesis. The 1H and 13C NMR spectra of the furanose-pyranose mixture of 4, after dissolution and equilibration in d(4)-methanol, were fully assigned allowing to expect that 4 could crystallise in the beta-pyranose form. New acetylation conditions able to trap this form were subsequently devised, allowing the isolation of 83% of pure 6beta by simple crystallisation, along with 9% of the 6beta/6alpha mixture. This represents a major advantage over the previously published procedure, especially on multigram scales. PMID- 12644381 TI - Carbohydrate esterase family 4 enzymes: substrate specificity. AB - The substrate specificity of selected enzymes classified under Carbohydrate Esterase family 4 (CE4) has been examined. Chitin deacetylase from Mucor rouxii and both a native and a truncated form of acetyl xylan esterase from Streptomyces lividans were found to be active on both xylan and several soluble chitinous substrates. Furthermore, the activities of all enzymes examined were significantly increased in the presence of Co(2+) when chitinous substrates were employed. However, the presence of this metal ion did not result in enhancing the activities of the enzymes when xylan was used as substrate. An acetyl xylan esterase from Bacillus pumilus, classified under Carbohydrate Esterase family 7, was found to be inactive towards all chitinous substrates tested. Finally, all enzymes examined were inactive towards cell wall peptidoglycan. PMID- 12644382 TI - Differentiation of embryonic stem cells for pharmacological studies on adipose cells. AB - The ongoing global explosion in the incidence of obesity has focused attention on the development of adipose cells. Severe obesity is the result of an increase in fat cell size in combination with increased fat cell number. New fat cells arise from a pre-existing pool of adipose stem cells that are present irrespective of age. The development of established preadipocyte cell lines has facilitated the study of different steps leading to terminal differentiation. However, these systems are limited for studying early events of differentiation as they represent cells which are already determined for the adipogenic lineage. In vitro differentiation of mouse embryonic stem (ES) cells towards the adipogenic lineage provides an alternative source of adipocytes for study in tissue culture and offers the possibility to investigate regulation of the first steps of adipose cell development. In this review, we describe the sequential requirement of retinoic acid and PPARgamma during adipogenesis in ES cells. Stimulation of ES cells with synthetic retinoids which are selective ligands of the retinoic acid receptor isotypes allowed the investigation of the contribution of the different retinoic receptors on the RA-dependent differentiation. The effects of thiazolidinediones, a new class of pharmacological agents used for the treatment of type 2 diabetes, and of statins, drugs used in therapy for lowering cholesterol, on the differentiation of ES cells into adipocytes or osteoblasts are described. Finally, we propose a model in which PPARgamma plays a key role in the decision of stem cells to undergo differentiation into adipocytes or osteoblasts, two closely related lineages. PMID- 12644383 TI - Pharmacological potential of embryonic stem cells. AB - Established embryonic stem (ES) cell lines have been at the forefront of approaches to understand gene function during embryogenesis and in adult vertebrate organisms, principally due to exploitation of two essential attributes; their pluripotency or ability to contribute to all three germinal layers and germ line in mice and their ease of genetic modification. Endeavours to routinely establish ES cells from species other than mice have met with limited success, although with rapid progress being made in our understanding of their basic cell biology, the regular derivation of lines from pre-implantation embryos will become easier for many species including humans. With a recent growing awareness of how these cells can be made to grow in an unlimited, but regulated manner plus how their fate can be directed or manipulated into diverse, mature phenotypes in culture, it has become clear that the biological resource offers additional attractive features applicable for future biomedical research and therapy. Advanced mouse ES-based technologies are being used in the industry for pharmaceutical discovery and development, while it is also anticipated that human ES cell reagents will revolutionise aspects of regenerative medicine. This review will summarise the advantages, potential and great hope for ES cell based systems in these contexts. PMID- 12644384 TI - Stem cells for cell therapy in Parkinson's disease. AB - Clinical studies with intrastriatal transplants of human embryonic mesencephalic tissue have provided proof-of-principle for the cell replacement strategy in Parkinson's disease (PD) patients. The grafted dopaminergic neurons can reinnervate the denervated striatum, restore regulated dopamine (DA) release and movement-related frontal cortical activation, and give rise to significant symptomatic relief. However, there are several problems linked to the use of primary embryonic tissue: (i) lack of sufficient amounts of tissue for transplantation in a large number of patients; (ii) variability of functional outcome, with some patients showing major improvement and others modest if any clinical benefit; (iii) occurrence of troublesome dyskinesias in a significant proportion of patients after transplantation. Stem cells could be useful to generate large numbers of DA neurons in standardized and quality-controlled preparations. So far, neurons with at least some dopaminergic characteristics have been generated from stem cells. However, in most cases their survival after grafting in animal PD models has been poor and it is also unclear if they function as normal mesencephalic DA neurons. For the development of a clinically useful cell therapy in PD, it is also necessary to define better criteria for patient selection and how graft placement should be optimized in each patient. Several scientific issues need to be addressed before stem cell-based therapies can be tested in PD patients. PMID- 12644385 TI - Neural stem cells: a pharmacological tool for brain diseases? AB - Stem cells are believed to provide a tool by which new cells and tissues can be made and by which damaged ones can be replaced or repaired. Over the past few years, the existence of a subset of stem cells has been documented in the fetal brain, therefore named neural stem cells (NSCs). To this regard, the more recent demonstration that similar cells are present in the adult mammalian brain and retain the capability to produce new neurons, has undermined the dogma that neurons are only generated during the fetal life and has stimulated investigations into the regulation and role of adult neurogenesis. Here, we will review the recent advancements on the biology of brain stem cells and discuss the mechanisms and drugs regulating adult neurogenesis, aiming at better estimating the possible future applications of NSCs for brain repair. PMID- 12644386 TI - Histamine in the control of gastric acid secretion: a topic review. AB - In this paper, the current knowledge about the role of histamine in the control of gastric acid secretion is reviewed. In particular, we focus this topic into three sections considering the recent insights on: histamine receptor subtypes involved in gastric acid secretion, the interplay between neuronal-hormonal paracrine pathways and the cerebral histaminergic control of gastric secretion. From the careful perusal of scientific literature, the fundamental role of histamine as local stimulator of gastric acid secretion via H(2) receptors is fairly confirmed while for the H(3) receptor only a minor modulating role is hypothesized. An undisputed function of ECL cells as controllable source of histamine within the so-called gastrin-ECL cell-parietal cell axis is generally proposed and the intriguing possibility of a remote control of gastric secretion via H(3) receptors is also suggested. PMID- 12644387 TI - Inhibition of excitotoxicity in cultured rat cortical neurons by a mixture of conjugated linoleic acid isomers. AB - Glutamate excitotoxicity, which is mediated by both N-methyl-D-aspartate (NMDA) and non-NMDA receptors, directly contributes to the neuronal cell loss associated with both acute insults and chronic neurodegenerative disorders. Conjugated linoleic acid (CLA) is a group of dienoic derivatives of linoleic acid shown to have anticarcinogenic and antioxidative activities. To evaluate the effect of a mixture of CLA isomers (cis-9, trans-11 and cis-10, trans-12 octadecadienoic acids) on glutamate- and NMDA-induced excitotoxicity, primary cultures of rat cortical neurons were treated for 15 min with 300 microM glutamate or NMDA in the presence of various concentrations of CLA. After the exposure, cell cultures were maintained at 37 degrees C for 18 h in minimum essential medium supplemented with glucose. Neuronal injury was measured by a colorimetric cell proliferation assay, and a qualitative assessment was made by phase-contrast microscopy. CLA inhibited glutamate- and NMDA-induced neuronal cell death in a concentration-dependent fashion with the most effective dose for neuroprotection being 500 microM. These results demonstrate that a mixture of CLA isomers exhibits protective action against glutamate- and NMDA-induced excitotoxicity. PMID- 12644388 TI - Molsidomine antagonizes L-NAME-induced acquisition deficits in a recognition memory task in the rat. AB - The present study was designed to investigate the role of nitric oxide (NO) on the acquisition of a recognition memory task in the rat. For this purpose, the effects on memory exerted by pre-training administration of the NO synthase inhibitor L-NAME (N(omega)-nitro-L-arginine methyl ester) and the NO donor molsidomine (N-[ethoxycarbonyl]-3-[4-morpholinosydnomine]) were assessed by using the object recognition task, a working memory paradigm based on the differential exploration of a new and familiar object. In a first dose-response study, it was found that L-NAME (10, 30, and 60 mg kg(-1), i.p.) at 30 but not at 10 mg kg(-1) disrupted animals performance, whereas the dose of 60 mg kg(-1) induced side effects. Molsidomine (2 and 4 mg kg(-1), i.p.) at 4 but not at 2 mg kg(-1), antagonized the L-NAME-induced performance deficits. These results indicate that NO is involved in the acquisition of a recognition memory task. PMID- 12644389 TI - Effects of erdosteine treatment against doxorubicin-induced toxicity through erythrocyte and plasma oxidant/antioxidant status in rats. AB - The clinical use of doxorubicin (Dxr), an antineoplastic agent, is limited by its extensive toxicity which is mostly mediated by oxidant injury. We have studied the effect of erdosteine, a mucolytic drug showing antioxidant properties, in preventing Dxr-toxicity to improve future Dxr therapy. Male Sprague-Dawley rats were divided into four groups. The first group that underwent no medication was accepted as control group; the second group was treated with a single i.p. injection of Dxr (20 mg kg(-1) b.wt.); the third group was treated with oral erdosteine alone (10 mg kg(-1) b.wt. day(-1) for 12 days); and in the last group erdosteine was administered starting before Dxr injection for 12 days. The thiobarbituric acid reactive substances (TBARS) level of Dxr group was higher in both plasma and erythrocyte than the other groups. In plasma and erythrocyte, the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were increased in Dxr plus erdosteine group in comparison with control group, and the activities of GSH-Px were increased in Dxr plus erdosteine group in comparison with Dxr group. The erythrocyte catalase (CAT) activity of Dxr plus erdosteine group was higher than control and Dxr groups. Plasma xanthine oxidase activities and nitric oxide (NO) levels were not significantly different between groups, however erythrocyte NO level of Dxr group was higher than control. In conclusion, Dxr administration resulted in increased lipid peroxidation in plasma as well as erythrocyte and erdosteine treatment helped to prevent oxidative injury by increasing antioxidant enzymes, especially SOD, GSH-Px and CAT, in rats. PMID- 12644390 TI - Anti-diabetic effects of Gymnema yunnanense extract. AB - In this study, we evaluated anti-hyperglycemic effect and body weight reduction activity of Gymnema yunnanense extract in obese ob/ob and diabetic db/db mice. Animals received daily intraperitoneal injections of the extract 100 mg/kg for 12 days. On Day 5, the extract-treated ob/ob mice had significantly lower fasting blood glucose levels compared to vehicle-treated mice (161+/-14.5mg/dl versus 238+/-21.5mg/dl, P<0.01). On Day 12, the extract-treated ob/ob mice had normal fasting blood glucose levels, compared with vehicle-treated mice (119+/-3.3mg/dl versus 240+/-12.9 mg/dl, P<0.01). Glucose tolerance improved significantly. This was demonstrated by overall glucose excursion calculated as area under the curve (AUC) during the 2h intraperitoneal glucose tolerance test (IPGTT), which decreased by approximately 22% (P<0.01) compared to vehicle-treated ob/ob mice. In addition, extract-treated ob/ob mice lost weight significantly from 51.7+/-1.9 g on Day 0 to 45.7+/-1.2g on Day 12 (P<0.05 compared to vehicle-treated mice). In db/db mice, after treatment with same dose of the extract, fasting blood glucose levels also decreased significantly from Day 0 of 247+/-13.9 mg/dl to Day 5 of 172+/-7.5mg/dl and to Day 12 of 190+/-2.7 mg/dl (both P<0.01 compared to vehicle treated group from Day 0 of 239+/-12.1mg/dl to Day 5 of 230+/-8.5mg/dl and Day 12 of 247+/-18.9 mg/dl, respectively). After 12 days of extract treatment, body weight in db/db mice reduced from 61.8+/-1.4 g on Day 0 to 59.8+/-1.1g on Day 12 (P<0.05). Our results support an in vivo anti-hyperglycemic and body weight reduction activity of G. yunnanense extract that may prove to be of clinical importance in improving the management of type 2 diabetes. PMID- 12644391 TI - The anti-inflammatory effects of the phosphodiesterase inhibitor pentoxifylline in the rat. AB - The present study aimed to evaluate the anti-inflammatory effects of pentoxifylline (PTX), a non-specific phosphodiesterase inhibitor in the rat. Acute inflammation was induced by subplantar injection of carrageenan (1%) in the rat hind paw. Results showed that intraperitoneal (i.p.) administration of PTX (36 or 72 mg kg(-1)) 30 min prior to carrageenan reduced the paw oedema response in dose-dependent manner with a maximal effect of 18.9 and 40.1%, respectively, at 2h post-carrageenan (P<0.001 and <0.001 at respective doses). Theophylline given at equimolar doses (29.9 or 45.8 mg kg(-1), i.p.) did not reduce the oedema response. With higher doses of PTX (144-300 mg kg(-1), i.p.) the anti-oedema effect of the drug was more pronounced, but mainly confined to the first 2h following carrageenan injection and decreasing rapidly thereafter. PTX (72 mg kg( 1), i.p.) given 30 min after carrageenan challenge reduced the oedema response by 24.7 and 26.2% at 1 and 2h after dosing (P<0.05 and <0.05, respectively). PTX (36 or 72 mg kg(-1), i.p.) co-administered with indomethacin (5 mg kg(-1), i.p.) 30 min before carrageenan had little modulatory effect on the anti-oedema effect of indomethacin, but the higher dose of PTX (144 mg kg(-1), i.p.) reduced the anti inflammatory effect of indomethacin by 24% at 4h post-carrageenan. PTX (72 mg kg( 1), i.p.) enhanced the anti-oedema effect of the selective COX-2 inhibitor celecoxib (33 mg kg(-1), i.p.) by 55.1% at 4h post-carrageenan. In contrast, the higher dose of PTX (144 mg kg(-1), i.p.) reduced the anti-oedema effect of celecoxib by 46.8% at 4h post-carrageenan. PTX (36 or 72 mg kg(-1)) enhanced the anti-oedema effect of dexamethasone (0.1 mg kg(-1)) with maximal effect of 76 and 104.8% at 2h post-carrageenan (P<0.01 and <0.01 for respective doses). PTX (0.6 mg per paw) given with carrageenan into the rat hind paw reduced the oedema response with a maximal effect of 33.4% at 1h following carrageenan. PTX (0.6 mg per paw) given in the contralateral hind paw reduced the carrageenan-induced paw oedema for 1h by 32.2%. Thus, PTX, when given at doses comparable to those used in man for treatment of circulatory disorders displayed anti-inflammatory in vivo and enhanced the anti-inflammatory effect of a selective COX-2 inhibitor or dexamethasone. PTX may have therapeutic potential as anti-inflammatory agent either alone or in combination with non-steroidal anti-inflammatory drugs or with steroids. There is also an intriguing possibility for the use of topical preparations for the management of local inflammatory conditions. PMID- 12644392 TI - Increases in insulin-like growth factor-1 level and peroxidative damage after gestational ethanol exposure in rats. AB - Ethanol exposure during pregnancy elicits profound detrimental developmental and behavioral effects such as reduced levels of insulin-like growth factor-1 (IGF-1) in the fetus. However, few reports have addressed its impact on postpartum dams. This study was designed to examine the influence of gestational ethanol exposure on postpartum maternal organ oxidative damage and IGF-1 level. Pregnant female rats were pair-fed from Day 2 of gestation until labor with control or ethanol (6.36% (v/v)) liquid diets and were sacrificed 6 weeks after parturition (ethanol withdrawn after parturition). There was no difference in body weight during or after the gestational period between the control and ethanol groups. Litter size was significantly less for ethanol-fed dams. One-week postnatal pup survival was significantly lower in the ethanol-fed (57.1%) than the control (97.8%) group. Liver and kidney tissue IGF-1 levels and mRNA were elevated in the ethanol-fed mothers, accompanied by hepatic but not renal oxidative damage, indicated by profound lipid peroxidation (measured by malondialdehyde (MDA) and 4 hydroxynonenal (4-HNE)) and protein carbonyl formation. The levels of glutathione (GSH), glutathione disulfide (GSSG) and GSH/GSSG ratios in liver and kidney were not different between the ethanol-fed and control dams. Collectively, these data suggest that gestational ethanol exposure may lead to postpartum oxidative organ damage and a possible compensatory increase in organ IGF-1 levels. PMID- 12644393 TI - Effect of melatonin on estrogen and progesterone receptors in relation to uterine contraction in rats. AB - The present study was designed to investigate the possible modulator effect of melatonin on uterine estrogen and progesterone receptors in rats as well as the uterine response to oxytocin. Non-pregnant rats were pretreated with melatonin in a dose of 0.8 mg kg(-1) per day for 15 consecutive days. Control animals received the vehicle. The uterus was dissected out and uterine contraction in one horne was recorded in vitro for each animal as a response to oxytocin (0.5 x 10(-11) to 2 x 10(-11)M). The other uterine horne was subjected to estrogen and progesterone receptors detection by immunohistochemical and image analysis techniques. The results reveal a significant reduction (59%) in the number of uterine estrogen receptors with concomitant increase in the progesterone receptors (53%) in melatonin-pretreated rats as compared to the control ones. In addition, our data show an inhibitory effect of melatonin on the uterine contraction as a response to oxytocin (0.5 x 10(-11), 1 x 10(-11), and 2 x 10(-11)M) amounting to 48, 77, and 59.5% reduction, respectively, in the amplitude of contraction as well as 62, 19.9, and 47% reduction in the area under the curve (AUC) of uterine contractions, respectively. The data, so far obtained, may indicate a possible relationship of melatonin-induced modulation of the number of estrogen and progesterone receptors and its inhibitory effect on uterine contraction. These findings merit further investigations on the possible beneficial role of melatonin in a plethora of hormone-dependent uterine disorders. PMID- 12644394 TI - Involvement of mitogen-activated protein kinases in Group B Streptococcus-induced macrophage apoptosis. AB - We previously demonstrated that Group B Streptococcus (GBS), a pathogen that causes serious neonatal infections, induces macrophage apoptosis by beta hemolysin to avoid the host immune response. GBS-induced macrophage apoptosis is characterized by a calcium increase and is caspase-independent. This study reports the involvement of c-Jun NH(2)-terminal kinase (JNK), p38 and extracellular signal-regulated kinase (ERK), three members of mitogen-activated protein kinases (MAPKs) family, in GBS-induced macrophage apoptosis. Our data indicate that during induction of apoptosis live GBS stimulates a strong persistent activation of JNK and p38 with concomitant inhibition of ERK. The time courses of MAPKs activation strongly correlate with GBS-induced macrophage apoptosis and are macrophage:GBS ratio-dependent. In fact, when GBS does not cause macrophage apoptosis, e.g. low macrophage:GBS ratio or non hemolytic GBS (gGBS), it induces a transient activation of JNK, p38, and ERK MAPKs. These latter results indicate that sustained and persistent activation of JNK and p38 and inhibition of ERK are involved in the GBS-induced macrophage apoptotic process and suggest that the time course and balance of MAPKs activation are critical for different macrophage responses to GBS (apoptosis versus antimicrobicidal activity). This study indicates a correlation between MAPKs activation and GBS-induced macrophage apoptosis. However, since neither ERK nor p38 inhibitors had an effect on GBS-induced apoptosis, their role in the complex signal network leading to GBS-induced macrophage apoptosis remains to be defined. PMID- 12644395 TI - Molecular phylogenetics and evolution of the endemic Hawaiian genus Adenophorus (Grammitidaceae). AB - Recent studies of the phylogeny of several groups of native Hawaiian vascular plants have led to significant insights into the origin and evolution of important elements of the Hawaiian flora. No groups of Hawaiian pteridophytes have been subjected previously to rigorous phylogenetic analysis. We conducted a molecular phylogenetic analysis of the endemic Hawaiian fern genus Adenophorus employing DNA sequence variation from three cpDNA fragments: rbcL, atpbeta, and the trnL-trnF intergenic spacer (IGS). In the phylogenetic analyses we employed maximum parsimony and Bayesian inference. Bayesian phylogenetic inference often provided stronger support for hypothetical relationships than did nonparametric bootstrap analyses. Although phylogenetic analyses of individual DNA fragments resulted in different patterns of relationships among species and varying levels of support for various clades, a combined analysis of all three sets of sequences produced one, strongly supported phylogenetic hypothesis. The primary features of that hypothesis are: (1) Adenophorus is monophyletic; (2) subgenus Oligadenus is paraphyletic; (3) the enigmatic endemic Hawaiian species Grammitis tenella is strongly supported as the sister taxon to Adenophorus; (4) highly divided leaf blades are evolutionarily derived in the group and simple leaves are ancestral; and, (5) the biogeographical origin of the common ancestor of the Adenophorus-G. tenella clade remains unresolved, although a neotropical origin seems most likely. PMID- 12644396 TI - Taxonomy of Iberian Hoplia (Col., Scarabaeoidea, Hopliinae) based on mtDNA analysis. AB - The morphology of some Hoplia species (Scarabaeoidea: Hopliinae) is so variable that parapatric populations have often been considered different species or subspecies. In this study we analyze the nucleotide sequences of a fragment of mitochondrial gene cytochrome c oxidase subunit I (COI) of six species and two subspecies of Palaearctic Hoplia to reexamine the species limits. Based on the analysis of sequences from COI and morphological and ecological observations, we consider Hoplia freyi Baraud to be a junior synonym of Hoplia chlorophana Erichson and H. philanthus ramburi Heyden to be a junior synonym of H. philanthus philanthus (Fuessly). However, complete resolution of relationships among H. philanthus subspecies requires the addition of sequences from genes evolving faster than COI. Phylogenetic relationships among the species studied are discussed. PMID- 12644397 TI - Molecular phylogeny of the gobioid fishes (Teleostei: Perciformes: Gobioidei). AB - The phylogeny of groups within Gobioidei is examined with molecular sequence data. Gobioidei is a speciose, morphologically diverse group of teleost fishes, most of which are small, benthic, and marine. Efforts to hypothesize relationships among the gobioid groups have been hampered by the prevalence of reductive evolution among goby species; such reduction can make identification of informative morphological characters particularly difficult. Gobies have been variously grouped into two to nine families, several with included subfamilies, but most existing taxonomies are not phylogenetic and few cladistic hypotheses of relationships among goby groups have been advanced. In this study, representatives of eight of the nine gobioid familes (Eleotridae, Odontobutidae, Xenisthmidae, Gobiidae, Kraemeriidae, Schindleriidae, Microdesmidae, and Ptereleotridae), selected to sample broadly from the range of goby diversity, were examined. Complete sequence from the mitochondrial ND1, ND2, and COI genes (3573 bp) was used in a cladistic parsimony analysis to hypothesize relationships among the gobioid groups. A single most parsimonious topology was obtained, with decay indices indicating strong support for most nodes. Major phylogenetic conclusions include that Xenisthmidae is part of Eleotridae, and Eleotridae is paraphyletic with respect to a clade composed of Gobiidae, Microdesmidae, Ptereleotridae, Kraemeriidae, and Schindleriidae. Within this five-family clade, two clades are recovered. One includes Gobionellinae, which is paraphyletic with respect to Kraemeriidae, Sicydiinae, Oxudercinae, and Amblyopinae. The other contains Gobiinae, also paraphyletic, and including Microdesmidae, Ptereleotridae, and Schindleriidae. Previous morphological evidence for goby groupings is discussed; the phylogenetic hypothesis indicates that the morphological reduction observed in many goby species has been derived several times independently. PMID- 12644398 TI - Phylogeny of the eelpout genus Lycodes (Pisces, Zoarcidae) as inferred from mitochondrial cytochrome b and 12S rDNA. AB - The bottom-dwelling and species-rich eelpout genus Lycodes Reinhardt has a great potential for the study of Arctic marine speciation. Subdivision of the genus has been based on single or few morphological characters (e.g., lateral line configuration) with contradicting results and phylogenetic approaches have not been attended. Here we present the first phylogenetic analysis of the genus employing DNA sequences of the mitochondrial genes cytochrome b and 12S rDNA (714 bp). The analysis with the two genes combined resulted in two equally parsimonious trees. In both cladograms most of the previously suggested subgroups are para- or polyphyletic, except for the so-called short-tailed Lycodes spp., with a short tail, a single mediolateral lateral line configuration and a shallow or filled otolith sulcus. The group of long-tailed Lycodes spp., with ventral or ventro-medio-lateral types of lateral line configuration and a deep otolith sulcus, appears to be paraphyletic, since Pacific and Atlantic species in this group are not each other's closest relatives. Thus, the short-tailed species are placed in a derived clade, indicating a secondary shortening of the tail, and a "slope to shore" type of evolution. This is not in accordance with earlier assumptions of the more elongate, deeper living species being the more derived. The basal position of long-tailed Pacific species supports earlier theories of Pacific origin of the genus/family. Small genetic differences between Arctic/Atlantic species indicate a rather recent radiation in these areas after the opening of the Bering Strait 3.0-3.5 million years ago. PMID- 12644400 TI - Higher-level systematics of rodents and divergence time estimates based on two congruent nuclear genes. AB - Phylogenetic analysis of over 4600 aligned nucleotide sequences from two nuclear genes, growth hormone receptor and BRCA1, provided congruent phylogenies depicting relationships among the major lineages of rodents. Separate and combined analyses resulted in five major conclusions: (1) strong support for a monophyletic Myodonta (containing the superfamilies Muroidea + Dipodoidea), with subfamily Gerbillinae being more closely related to Murinae than is Sigmodontinae; (2) a sister-group relationship between the family Castoridae and the superfamily Geomyoidea; (3) monophyly of Ctenohystrica (containing the suborders Sciuravida and Hystricognatha); (4) a near polytomy among Myodonta (suborder Myomorpha), Pedetes (family Pedetidae, suborder Anomaluromorpha), Castoridae (suborder Sciuromorpha) + Geomyoidea (suborder Myomorpha), and Ctenohystrica; and (5) basal position of a monophyletic group containing Graphiurus (family Gliridae, suborder Myomorpha) + two members of the Sciuromorpha (Sciuridae + Aplodontidae). Divergence dates among rodents and primates were also estimated using the combined data. Applying a global molecular clock and a primate calibration point, divergence dates among rodents exceeded fossil-based dates but were generally compatible with other molecule-based dates estimated under similar conditions. However, when a relaxed molecular clock was applied, estimated divergence dates were highly compatible with the fossil record. PMID- 12644399 TI - Extensive mtDNA variation within the yellow-pine chipmunk, Tamias amoenus (Rodentia: Sciuridae), and phylogeographic inferences for northwest North America. AB - The yellow-pine chipmunk, Tamias amoenus, is common in xerophytic forests throughout much of northwest North America. We analyzed cytochrome b sequence variation from 155 individuals representing 57 localities across the distribution of T. amoenus including 10 additional species of Tamias. Maximum likelihood and parsimony tree estimation methods were used in conjunction with nested clade analysis to infer both deep and population-level processes. Our results indicate that two currently recognized subspecies of T. amoenus (T. a. canicaudus and T. a. cratericus) are not nested within other samples of T. amoenus. Maximum uncorrected levels of intraspecific sequence divergence within remaining samples of T. amoenus are >7%. Substantial geographic variation is characterized by 12 well-supported clades that correspond to distinct mountain ranges, but do not necessarily follow existing subspecific taxonomy. Significant association between geography and genealogy was detected within many of these clades and can be attributed to different population-level processes including past fragmentation, recent range expansion, and isolation by distance. PMID- 12644401 TI - Mitochondrial cytochrome b of the Lyakhov mammoth (Proboscidea, Mammalia): new data and phylogenetic analyses of Elephantidae. AB - The phylogenetic relationships between recent Elephantidae (Proboscidea, Mammalia), that is to say extant elephants (Asian and African) and extinct woolly mammoth, have remained unclear to date. The prevailing morphological scheme (mammoth grouped with Asian elephant) is either supported or questioned by the molecular results. Recently, the monophyly of woolly mammoths on mitochondrial grounds has been demonstrated (Thomas, et al., 2000), but it conflicts with previous studies (Barriel et al., 1999; Derenko et al., 1997). Here, we report the partial sequencing of two mitochondrial genes: 128 bp of 12S rDNA and 561 bp of cytochrome b for the Lyakhov mammoth, a 49,000-year-old Siberian individual. We use the most comprehensive sample of mammoth (11 sequences) to determine whether the sequences achieved by former studies were congruent or not. The monophyly of a major subset of mammoths sequences (including ours) is recovered. Such a result is assumed to be a good criterion for ascertaining the origin of ancient DNA. Our sequence is incongruent with that of Yang et al. (1996), though obtained for the same individual. As far as the latter sequence is concerned, a contamination by non-identified exogenous DNA is suspected. The robustness and reliability of the sister group relation between Mammuthus primigenius and Loxodonta africana are examined: down-weighting saturated substitutions has no impact on the topology; analyzing data partitions proves that the support of this clade can be assigned to the most conservative phylogenetic signal; insufficient taxonomic and/or characters sampling contributed to former discordant conclusions. We therefore assume the monophyly of "real mammoth sequences" and the (Mammuthus, Loxodonta) clade. PMID- 12644403 TI - The effects of increasing genetic distance on alignment of, and tree construction from, rDNA internal transcribed spacer sequences. AB - We examined how alignment of internal transcribed spacers of rDNA in fungi and plants changes with increasing genetic distance by successive removal of sequences from each data set followed by realignment and phylogenetic analysis. Increasing genetic distance can negatively affect phylogenetic reconstruction in two ways. First, it may cause errors in the alignment and therefore the homology hypotheses of the sequence characters. Second, it may cause errors in the homology assessments of character states because of multiple hits on individual branches. These two causes of error in phylogenetic inference were distinguished from one another in our analysis. The errors in alignment caused by increasing genetic distance were primarily due to inserting too few gaps and inserting gaps at the wrong positions. Errors in tree resolution, topology, and/or branch support values were more often caused by multiple hits than by misaligned positions. This suggests that increasing genetic distance negatively affects our primary homology assessments of character states more severely than our primary homology assessments of characters. We suggest that increasing taxon sampling with the aim of subdividing long branches is a strategy for obtaining reliable alignments. PMID- 12644402 TI - Relationships and origin of endemic Lake Baikal gastropods (Caenogastropoda: Rissooidea) based on mitochondrial DNA sequences. AB - The phylogenetic relationships and the origin of two groups of rissooid freshwater snails endemic to Lake Baikal were investigated using partial mitochondrial COI, 12S rDNA, and 16S rDNA sequences. The Baikalian Benedictiinae proved to be closely related to the Lithoglyphinae. According to a molecular clock estimate the two groups diverged in the Paleogene. The Benedictiinae might have evolved autochthonously in precursors of Lake Baikal. The Baikalian Baicaliidae are probably most closely related to the Amnicolidae and the Bithyniidae. These groups diverged at the latest during the Cretaceous. Thus the origin of the Baicaliidae predates the origin of the Baikal rift zone. The Baicaliidae evolved probably in other Central Asian freshwater reservoirs. However, the radiation of the extant Baicaliidae only started in the Neogene and might have occurred autochthonously in Lake Baikal. The conchological similarity of the Baicaliidae and the Pyrgulidae is due to convergence. The Pyrgulidae diverged from the common stem lineage of the other hydrobiid families at the latest in the Jurassic. The Bithyniidae is derived from hydrobiids and is related to the Amnicolidae. PMID- 12644404 TI - Molecular systematics and phylogeography of Amazonian poison frogs of the genus Dendrobates. AB - The study of Amazonian biodiversity requires detailed knowledge of the phylogenetic relationships of closely related taxa distributed across Amazonia. The Amazonian poison frogs of the genus Dendrobates have undergone many taxonomic revisions, but the phylogenetic relationships within this group remain poorly understood. Most previous classifications were based on morphology and skin toxin analyses, with limited use of DNA sequence data. Using mtDNA sequence data from four gene regions (cytochrome b, cytochrome oxidase I, 16S rRNA, and 12S rRNA), we present a molecular phylogenetic analysis of the evolutionary relationships within a representative group of Amazonian Dendrobates. We use the resulting phylogenetic hypothesis to investigate different biogeographic hypotheses concerning genetic divergence and species diversity in Amazonia. The results of the analysis support the presence of ancient paleogeographic barriers to gene flow between eastern and western Amazonia, and indicate substantial genetic divergence between species found in the northern and southern regions of western Amazonia. PMID- 12644405 TI - Molecular systematics of the South American caviomorph rodents: relationships among species and genera in the family Octodontidae. AB - Nucleotide sequences from mitochondrial (12S rRNA) and nuclear (growth hormone receptor) genes were used to investigate phylogenetic relationships among South American hystricognath rodents of the superfamily Octodontoidea, with special emphasis on the family Octodontidae. Relationships among most taxa were well resolved by a combined analysis of both genes, and the molecular phylogeny was used to address several long-standing phylogenetic problems. The family Abrocomidae was the most basal lineage within the superfamily Octodontoidea, sensu stricto, and the family Ctenomyidae was sister to the family Octodontidae, followed by a monophyletic group containing the families Myocastoridae and Echimyidae. A basic dichotomy was observed within the family Octodontidae. The Argentine desert specialists, Tympanoctomys and Octomys, grouped separate from Octodontomys, which was sister to a clade containing a monophyletic Octodon and a clade represented by species of Aconaemys and Spalacopus. Aconaemys was paraphyletic relative to Spalacopus. The phylogeny was used as an interpretive framework for an examination of variation in several non-molecular characters. The primitive diploid number for most of the octodontoids was determined to be between 46 and 56, and the primitive genome size 8.2 pg. Members of the Octodontidae appeared to be derived from an ancestral stock occupying lower elevations in scrub habitat. Furthermore, estimates of divergence time from the molecular data provided a temporal perspective for changes in plant communities, which demonstrated turnover and diversification in response to climatic and geologic events occurring in the Miocene through the Pleistocene. PMID- 12644406 TI - Molecular cladistic markers in New World monkey phylogeny (Platyrrhini, Primates). AB - Transpositions of primate-specific Alu elements were applied as molecular cladistic markers in a phylogenetic analysis of South American primates. Seventy four human and platyrrhine loci containing intronic Alu elements were PCR screened in various New World monkeys and the human outgroup to detect the presence of orthologous retrotransposons informative of New World monkey phylogeny. Six loci revealed size polymorphism in the amplification pattern, indicating a shared derived character state due to the presence of orthologous Alu elements confirmed by subsequent sequencing. Three markers corroborate (1) New World monkey monophyly and one marker supports each of the following callitrichine relationships: (2) Callithrix and Cebuella are more closely related to each other than to any other callitrichine, (3) the callitrichines form a monophyletic clade including Callimico, and (4) the next living relatives to the callitrichines are Cebus, Saimiri, and Aotus. PMID- 12644407 TI - Sequence and phylogenetic analysis of the complete mitochondrial genome of the flour beetle Tribolium castanaeum. AB - We describe the first complete mitochondrial genome sequence from a representative of the insect order Coleoptera, the flour beetle Tribolium castaneum. The 15,881 bp long Tribolium mitochondrial genome encodes 13 putative proteins, two ribosomal RNAs and 22 tRNAs canonical for animal mitochondrial genomes. Their arrangement is identical to that in Drosophila melanogaster, which is considered ancestral for insects and crustaceans (Boore et al., 1998; Hwang, et al., 2001a). Nucleotide composition, amino acid composition, and codon usage fall within the range of values observed in other insect mitochondrial genomes. Most notable features are the use of TCT as tRNA(Ser(AGN)) anticodon instead of GCT, which is used in most other arthropod species, and the relative scarcity of special sequence motifs in the 1431 bp long control region. Phylogenetic analysis confirmed resolving power in the conserved regions of the mitochondrial proteome regarding diversification events, which predate the emergence of pterygote insects, while little resolution was obtained at the level of basal perygote diversification. The partition of faster evolving amino acid sites harbored strong support for joining Lepidoptera with Diptera, which is consistent with a monophyletic Mecopterida. PMID- 12644408 TI - Phylogenetic and genomic analysis of the complete mitochondrial DNA sequence of the spotted asparagus beetle Crioceris duodecimpunctata. AB - We report the complete mitochondrial DNA sequence of the spotted asparagus beetle, Crioceris duodecimpunctata. The genome complement, gene order, and nucleotide composition of this beetle's mitochondrial genome were found to be typical of those reported for other insects. Unusual features of this genome include the substitution of UCU for GCU as the anticodon for tRNA(Ser), an unusual TpsiC loop for the tRNA(Ile) gene, and the identification of a putative ATT start codon for cox1. The utility of complete mitochondrial genome data for phylogenetic inference of the insect orders was tested, and compared to that of cox1 and combined mitochondrial ribosomal DNA sequences. Even though the number of insect orders represented by complete mitochondrial genomes is still limited, several well-established relationships are evident in the phylogenetic analysis of the complete sequences. Monophyly of the orders Diptera, Lepidoptera, and Coleoptera were consistently recovered. Monophyly of the Holometabola was also observed in some (though not all) analyses. The accumulation of complete mitochondrial sequences from a broader array of insect orders holds the promise of clarifying the early diversification of insects. PMID- 12644410 TI - Near-patient testing--point-of-care or point of costs and convenience? PMID- 12644411 TI - Anaesthesia for the diabetic patient. PMID- 12644412 TI - Hyperglycaemia and the lung. PMID- 12644413 TI - Clonidine premedication improves metabolic control in type 2 diabetic patients during ophthalmic surgery. AB - BACKGROUND: In stressful conditions, increasing blood glucose concentrations are closely related to an increase in catecholamines and cortisol release. Clonidine, a centrally acting alpha(2)-adrenoceptor agonist, has neuroendocrine effects, including inhibition of sympathoadrenal activity. We therefore evaluated the effect of clonidine on blood glucose control and insulin requirements during ophthalmic surgery when given as premedication in type 2 diabetic patients. METHODS: After randomization, patients were premedicated with clonidine or flunitrazepam (control). Patients were given insulin by continuous i.v. infusion to maintain blood glucose in the range 5.5-11.1 mmol litre(-1). Blood glucose concentrations were measured every 15 min during surgery, and hourly for 6 h after surgery. Plasma C-peptide and counter-regulatory hormones were also measured. RESULTS: Glycaemia was significantly lower in the clonidine group (P<0.01) and the median amount of insulin administered was significantly reduced: clonidine group 9.0 (interquartile range 5.1) units; control 18.6 (10.2) units; P<0.01). Plasma catecholamine concentrations were lower in patients given clonidine (P<0.05) but there was no difference in cortisol concentrations. CONCLUSION: Premedication of type 2 diabetic patients with clonidine 90 min before surgery improves blood glucose control and decreases insulin requirements during ophthalmic surgery. PMID- 12644414 TI - Adding lactate to the prime solution during hypothermic cardiopulmonary bypass: a quantitative acid-base analysis. AB - BACKGROUND: The effect of adding lactate to the cardiopulmonary bypass (CPB) prime was investigated using Stewart's quantitative acid-base approach. According to this quantitative model, serum pH and bicarbonate are determined by three independent factors: the partial pressure of carbon dioxide (PCO(2)), the total concentration of weak acids (e.g. albumin), and the strong ion difference. The apparent strong ion difference is calculated as the sum of sodium, potassium, magnesium and calcium minus chloride concentrations. The pH decreases with a smaller strong ion difference and vice versa. METHODS: Twenty patients scheduled for coronary surgery were studied prospectively. All patients were treated identically, except for the prime, which either contained lactate or was lactate free. Just before bypass and before coming off bypass, haemoglobin, glucose, plasma osmolality and colloid osmotic pressure were determined; albumin, lactate, sodium, potassium, ionized calcium, magnesium, phosphate, arterial pH, PCO(2), bicarbonate, and base excess were measured for use in Stewart's analysis. RESULTS: Metabolic acidosis had resolved by the end of bypass with the lactated prime. Although the strong ion gap (apparent minus effective strong ion difference) increased significantly in both groups, its composition differed significantly between the groups. The Stewart technique detected polyanionic gelatin as a weak acid component contributing to the unidentified anion fraction. Colloid osmotic pressure was maintained in both groups. CONCLUSION: Exogenous lactate attenuates acidosis related to CPB. The oncotic and weak acid deficits produced by hypoalbuminaemia may be compensated for temporarily during CPB by polyanionic synthetic colloids such as succinylated gelatin. PMID- 12644415 TI - Transient hyperaemic response to assess vascular reactivity of skin: effect of locally iontophoresed acetylcholine, bradykinin, epinephrine and phenylephrine. AB - BACKGROUND: Recently, the transient hyperaemic response (THR) to brief compression (20 s) of the brachial artery has been described as a way to assess vascular reactivity of the forearm skin. We studied the effects of locally iontophoresed vasoactive agents on this response in 20 male volunteers. METHODS: An iontophoresis chamber attached to the anterior forearm permitted simultaneous administration of drugs by iontophoresis and measurement of skin blood flow-flux by laser Doppler probe. Three THR tests were performed before and after iontophoresis by compressing the brachial artery with digital pressure for 20 s and then releasing. The following were iontophoresed: saline 0.9% (iontophoresis vehicle control), acetylcholine, bradykinin, epinephrine and phenylephrine. The THR ratio (THRR) was calculated as F2/F1 where F1 was baseline blood flow-flux immediately before compression and F2 was peak blood flow-flux after release. RESULTS: When compared with saline 0.9%, acetylcholine and bradykinin increased median F1 from 9.2 (range 5.2-23.8) to 22.1 (8.7-61.5) and from 4.8 (3.0-23.2) to 15.0 (2.5-31.8), respectively, and reduced THRR from 1.26 (1.07-2.2) to 0.99 (0.93-1.04) and from 1.63 (1.06-2.58) to 1.09 (0.93-1.19), respectively. Epinephrine, but not phenylephrine, caused a significant reduction in F1 from 9.2 (5.2-23.8) to 4.0 (1.5-22.3). Neither epinephrine nor phenylephrine had significant effect on THRR. CONCLUSIONS: Iontophoresed acetylcholine and bradykinin significantly increase the flow-flux and impair THR in forearm skin, further validating the concept that THR represents true vasodilatation during arterial occlusion. In addition, iontophoresis of vasoconstrictors does not appear to have any consistent effect. PMID- 12644416 TI - How high do the subclavian arteries ascend into the neck? A population study using magnetic resonance imaging. AB - BACKGROUND: The relationship between the larynx and the subclavian arteries was studied in a series of magnetic resonance images (MRIs) from 50 patients without neck pathology. METHODS: The vertical distances of the excursion of the subclavian arteries into the neck was measured, as was the distance from the cricoid cartilage to the highest point of this excursion. Statistical analysis allows the probability of any given cricoid-subclavian distance occurring in the population to be estimated. RESULTS: The mean (SD) excursion of the right subclavian artery above the clavicle was 10.4 (11.4) mm. The mean (SD) distance from the cricoid cartilage to the right subclavian artery was 30.6 (14.3) mm, and the data showed a high degree of variance. There was a linear relationship between neck length and cricoid-subclavian distance (r=0.58), which explained some of the variance in the latter, but there was also wide individual variance, which was independent of this regression. CONCLUSIONS: When performing a percutaneous tracheostomy, a 'safe' distance between the incision site and subclavian artery cannot be assumed or reliably predicted from the neck length. PMID- 12644417 TI - Comparison of two Macintosh laryngoscope blades in 300 patients. AB - BACKGROUND: There are two forms of Macintosh laryngoscope blade. Compared with the standard blade, the English blade is longer, its curve is more continuous across the entire length of the blade, the flange of the blade continues much closer to the blade tip, and the height of the flange is shorter. METHOD: We studied 300 patients to compare the ease of laryngoscopy with each type of Macintosh laryngoscope blade. In a random crossover design, after induction of anaesthesia and neuromuscular block, the two blades were inserted in turn, and the views of the glottis at laryngoscopy (Cormack and Lehane scores) were compared. RESULTS: There was a difference in the view of the glottis in 80 patients. Among these patients, the view was better for the English blade for 63 patients and the standard blade was better for 17 patients. Laryngoscopy was difficult (grade 3 or 4) for at least one blade in 42 of 300 patients (14%). In these 42 patients, there was a difference in the score between the blades in 28 patients; the view was better for the English blade in 25 patients (60%) and for the standard blade in three patients (7%). The view was significantly better for the English blade than for the standard blade (P<0.001; 95% confidence interval 45-74%). CONCLUSIONS: In patients in whom laryngoscopy was unexpectedly difficult, the English blade provided a better glottic view significantly more frequently than the standard blade. PMID- 12644418 TI - Propofol and halothane versus sevoflurane in paediatric day-case surgery: induction and recovery characteristics. AB - BACKGROUND: The aim of this study was to compare the induction and recovery characteristics associated with propofol induction and halothane maintenance with sevoflurane anaesthesia in paediatric day surgery. METHODS: In total, 322 children were assigned randomly to i.v. propofol induction and halothane/nitrous oxide maintenance or sevoflurane/nitrous oxide alone. The patients' age, sex, and type of surgery were recorded, as were the times required for anaesthetic induction, maintenance, recovery and time to discharge home. Postoperative nausea and vomiting, and the incidence of adverse events during induction and recovery were also noted. RESULTS: No significant differences were detected in age, sex, type of surgery performed or intraoperative opioid administration. Excitatory movement was more common during induction with sevoflurane. The mean time required for induction with propofol was 3.1 min compared with 5 min in the sevoflurane group (P<0.001). The recovery time was shorter in the sevoflurane group compared with propofol/halothane (23.2 vs 26.4 min, P<0.002). The incidence of delirium in recovery was greater in the sevoflurane group (P<0.001). There was no difference between groups in the time spent on the postoperative ward before discharge home. On the postoperative ward the incidence of both nausea and vomiting was significantly higher in the sevoflurane group (P=0.034). Five children were admitted to hospital overnight, none for anaesthetic reasons. CONCLUSIONS: The increased incidence of adverse events during induction, postoperative nausea and vomiting and postoperative delirium in the sevoflurane group suggests that sevoflurane is not ideal as a sole agent for paediatric day case anaesthesia. PMID- 12644419 TI - Lumbar epidural fentanyl: segmental spread and effect on temporal summation and muscle pain. AB - BACKGROUND: Despite extensive use, different aspects of the pharmacological action of epidural fentanyl have not been clarified. We applied a multi-modal sensory test procedure to investigate the effect of epidural fentanyl on segmental spread, temporal summation (as a measure for short-lasting central hyperexcitability) and muscle pain. METHODS: Thirty patients received either placebo, 50 or 100 micro g single dose of fentanyl epidurally (L2-3), in a randomized, double-blind fashion. Heat pain tolerance thresholds at eight dermatomes from S1 to fifth cranial nerve (assessment of segmental spread), pain threshold to transcutaneous repeated electrical stimulation of the sural nerve (assessment of temporal summation) and pain intensity after injection of hypertonic saline into the tibialis anterior muscle (assessment of muscle pain) were recorded. RESULTS: Fentanyl 100 micro g, but not 50 micro g, produced analgesia to heat stimulation only at L2. Surprisingly, no effect at S1 was detected. Both fentanyl doses significantly increased temporal summation threshold and decreased muscle pain intensity. CONCLUSIONS: The findings suggest that a single lumbar epidural dose of fentanyl should be injected at the spinal interspace corresponding to the dermatomal site of pain. Increased effect on L2 compared with S1 suggests that drug effect on spinal nerve roots and binding to opioid receptors on the dorsal root ganglia may be more important than traditionally believed for the segmental effect of epidurally injected fentanyl. Epidural fentanyl increases temporal summation threshold and could therefore contribute to prevention and treatment of central hypersensitivity states. I.M. injection of hypertonic saline is a sensitive technique for detecting the analgesic action of epidural opioids. PMID- 12644420 TI - Immediate extubation and epidural analgesia allow safe management in a high dependency unit after two-stage oesophagectomy. Results of eight years of experience in a specialized upper gastrointestinal unit in a district general hospital. AB - BACKGROUND: The perioperative management of two-stage oesophagectomy has not been standardized and the prevailing practice regarding the timing of extubation after the procedure varies. This audit has evaluated the outcome, in particular the respiratory morbidity and mortality, after immediate extubation in patients who have had thoracic epidural analgesia. METHODS: All the patients who underwent two stage oesophagectomy by a single specialist upper gastrointestinal surgeon were recorded both retrospectively (1993-1999) and prospectively (1999-2001). Physical characteristics, comorbid factors, anaesthetic management and postoperative events were recorded on a computer database. Analysis was undertaken to evaluate the morbidity and mortality, in particular the need for reventilation and transfer to the ITU. RESULTS: Seventy-six patients underwent two-stage oesophagectomy between 1993 and 2001. Seventy-three (96%) patients were extubated in theatre and transferred to a high-dependency bed. Three were ventilated electively and extubated within 36 h and made an uncomplicated recovery. Seven (10%) of the immediately extubated patients subsequently needed admission to the ICU and reventilation. Sixty-seven patients had effective epidural analgesia and nine needed i.v. morphine by patient-controlled analgesia. The 30-day or in hospital mortality was 2.6% (2 of 76). A further two patients died within 90 days, but after discharge. Respiratory complications were responsible for half of the overall morbidity (44.7%). Respiratory failure occurred in 6.5% (5 of 76) and acute respiratory distress syndrome in 2.6% (2 of 76). Both the in-hospital deaths occurred in patients requiring reventilation and resulted from respiratory complications. The following factors were found to be significant in the reventilated patients: duration of one-lung ventilation; forced expiratory volume in the first second; and ratio of forced expiratory volume in the first second/forced vital capacity. CONCLUSIONS: Immediate extubation after two-stage oesophagectomy in patients with thoracic epidural analgesia is safe and associated with low morbidity and mortality. Patients can be managed in a high dependency unit, thus avoiding the need for intensive care. This has cost-saving and logistical implications. PMID- 12644422 TI - Caudal additives in children--solutions or problems? PMID- 12644421 TI - Monitoring of neuromuscular block after administration of vecuronium in patients with diabetes mellitus. AB - BACKGROUND: We studied the supramaximal current for ulnar nerve stimulation during electromyographic monitoring of onset and recovery of neuromuscular block using a neuromuscular transmission module (M-NMT Module, Datex-Ohmeda) in patients with Type 2 diabetes undergoing anaesthesia with nitrous oxide, oxygen, isoflurane and fentanyl. METHODS: Thirty-six diabetic patients were randomly assigned to a post-tetanic count (PTC) group (n=17) or train-of-four (TOF) group (n=19). In addition, 30 non-diabetic patients were divided into control PTC (n=15) and TOF groups (n=15). RESULTS: In the diabetic patients (diabetes PTC and diabetes TOF groups), the mean supramaximal stimulating current was significantly higher than in the non-diabetic patients (control PTC and TOF groups) (50.5 (SD 14.1) vs 33.4 (6.1) mA, P<0.01). Onset of neuromuscular block (time to disappearance of T1) after vecuronium 0.1 mg kg(-1) in the diabetic patients did not differ significantly from that in the non-diabetic patients (276 (77) vs 244 (44) s, P=0.055). Time to return of PTC1 did not differ significantly between the diabetes and control PTC groups (21.0 (12.1) vs 15.7 (5.0) min, P=0.126). Times to return of T1 and T4 in the diabetes TOF group were significantly longer than in the control TOF group (T1: 37.5 (15.2) vs 25.7 (7.6) min, P=0.01; T4: 61.4 (23.7) vs 43.5 (11.4) min, P=0.01). During recovery, PTC and T4/T1 in the diabetes PTC and TOF groups were similar to those in the control PTC and TOF groups, respectively. T1/T0 in the diabetes TOF group was significantly less than in the control TOF group, 80-120 min after vecuronium (P<0.05). CONCLUSIONS: In diabetic patients, supramaximal current is higher than in non-diabetic patients. After vecuronium, onset of neuromuscular block and recovery of PTC or T4/T1 are not altered, but time to return of T1 or T4, and recovery of T1/T0 are delayed in diabetic patients. PMID- 12644423 TI - Near-patient testing for coagulopathy after cardiac surgery. AB - BACKGROUND: We assessed the accuracy and precision of a new near-patient testing system (Hemochron Response) by measuring prothrombin time and activated partial thromboplastin time (PT and APTT) in 50 patients undergoing cardiac surgery using cardiopulmonary bypass and comparing the results with laboratory assays. METHODS: Blood samples were taken at the beginning of surgery and the PT and APTT was measured both in the laboratory and by the Hemochron Response. The tests were repeated 30 min after reversal of heparin with protamine. RESULTS: Before bypass, the bias for PT was only +0.34, with small 95% limits of agreement. Making the same measurements after bypass, the Hemochron Response under-read and the bias was -3.27, with an increase of the 95% limits of agreement. With the APTT, the bias and the 95% limits of agreement were greater before bypass, and became even wider after bypass. CONCLUSIONS: We found good agreement in the PT and clinically acceptable levels of agreement in the APTT during the pre-bypass period. After bypass, bias became greater for both PT and APTT and the limits of agreement could be clinically unacceptable. PMID- 12644424 TI - Effects of halothane on action potential configuration in sub-endocardial and sub epicardial myocytes from normotensive and hypertensive rat left ventricle. AB - BACKGROUND: Halothane shortens ventricular action potential duration (APD), as a consequence of its inhibitory effects on a variety of membrane currents, an effect that is greater in sub-endocardial than sub-epicardial myocytes. In hypertrophied ventricle, APD is prolonged as a consequence of electrical remodelling. In this study, we compared the effects of halothane on transmural APD in myocytes from normal and hypertrophied ventricle. METHODS: Myocytes were isolated from the sub-endocardium and sub-epicardium of the left ventricle of spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) rats. Action potentials were recorded before, during, and after a 1-min exposure to 0.6 mM halothane and APD measured from the peak of the action potential to repolarization at -50 mV (APD(-50 mV)). Data are presented as mean (SEM). RESULTS: In WKY myocytes, halothane reduced APD(-50 mV) from 21 (2) to 18 (2) ms (P<0.001, n=15) in sub-epicardial myocytes but abbreviated APD(-50 mV) to a greater extent in sub-endocardial myocytes (37 (4) to 28 (3) ms; P<0.001, n=14). In SHR myocytes, APD(-50 mV) values were prolonged compared with WKY and APD(-50 mV) was reduced by halothane from 36 (6) to 27 (4) ms (P<0.016) and from 77 (10) to 38 (4) ms (P<0.001) in sub-epicardial and sub-endocardial myocytes, respectively. CONCLUSIONS: In the SHR, hypertrophic remodelling was not homogeneous; APD(-50 mV) was prolonged to a greater extent in sub-endocardial than sub-epicardial cells. Halothane reduced APD to a greater extent in sub endocardium than sub-epicardium in both WKY and SHR but this effect was larger proportionately in SHR myocytes. The transmural gradient of repolarization was reduced in WKY and effectively abolished in SHR by halothane, which might disturb normal ventricular repolarization. PMID- 12644425 TI - Pharmacological effects of intravenous melatonin: comparative studies with thiopental and propofol. AB - BACKGROUND: Possible utility of high-dose i.v. melatonin as an anaesthetic adjuvant has not been studied. This study compared its effects with thiopental and propofol. METHODS: Sprague Dawley rats were assigned to receive bolus or cumulative i.v. doses of melatonin, thiopental or propofol. Righting reflex, hindpaw withdrawal to a noxious stimulus, response to tail clamping and haemodynamic effects were assessed. RESULTS: Melatonin caused a dose-dependent increase in paw withdrawal threshold and the percent of rats displaying loss of the righting reflex. Melatonin was comparable to thiopental and propofol in terms of its rapid onset of hypnosis. The mean ED(50) values for loss of righting reflex were 5.4 (SEM 1.2), 12.5 (1.1) and 178 (1.1) mg kg(-1) for propofol, thiopental and melatonin, respectively. The percent of rats displaying loss of response to tail clamping was greater with propofol than with melatonin (P<0.05). Haemodynamic changes produced by melatonin or propofol were similar in onset and magnitude. CONCLUSIONS: I.V. melatonin can exert hypnotic effects similar to those observed with thiopental and propofol. Melatonin exhibited significant antinociceptive effects but was less effective in abolishing the response to tail clamping. PMID- 12644426 TI - Effect of continuous low-dose intravenous diltiazem on epidural fentanyl analgesia after lower abdominal surgery. AB - BACKGROUND: The postoperative opioid-sparing effects of systemic L-type calcium channel blockers are controversial. We investigated whether the postoperative analgesic effect of epidural fentanyl was enhanced by i.v. infusion of diltiazem at a rate that would minimize any cardiovascular depressant effect. METHODS: After elective lower abdominal gynaecological surgery, 30 patients were randomized to receive continuous i.v. diltiazem 1 micro g kg(-1) min(-1) (diltiazem group) or the same volume of saline (control group) for 24 h. Cumulative postoperative epidural fentanyl consumption, visual analogue scale (VAS) scores and verbal rating scores (VRS) at rest and during mobilization, sedation scores, incidence of side-effects and overall patient satisfaction were assessed. RESULTS: There was no significant difference in cumulative epidural fentanyl consumption between the groups at any period. Although there were no statistically significant differences in VAS scores, VRS, sedation scores, incidence of side-effects and overall patient satisfaction, there was a trend to an increased incidence of nausea in the diltiazem group. CONCLUSIONS: Continuous i.v. infusion of diltiazem did not reduce epidural fentanyl consumption when administered at dosages having minimal haemodynamic depressant effects. PMID- 12644427 TI - Could ultrasonography be used by an anaesthetist to identify a specified lumbar interspace before spinal anaesthesia? AB - BACKGROUND: Insertion of a needle into the lumbar subarachnoid space may cause damage to the spinal cord. Current techniques to identify a safe interspace have limitations. Ultrasound was investigated as a means to improve anatomical accuracy. METHODS: Seventeen patients attending for elective magnetic resonance imaging (MRI) of the spine were studied. Ultrasonic identification of the L3-4 interspace was attempted by an anaesthetist and a marker was placed. A radiologist identified the anatomical location of the marker on the MRI scan. RESULTS: Thirteen out of 17 markers were at the L3-4 interspace; four were at the L2-3 interspace. CONCLUSIONS: These results suggest that ultrasonography may be a useful adjunct to safe subarachnoid anaesthesia. PMID- 12644428 TI - Comparison of L-bupivacaine 0.75% and lidocaine 2% with bupivacaine 0.75% and lidocaine 2% for peribulbar anaesthesia. AB - BACKGROUND: L-Bupivacaine has a safer side-effect profile than bupivacaine. We compared the efficacy of a mixture of L-bupivacaine 0.75% and lidocaine 2% with bupivacaine 0.75% and lidocaine 2% for peribulbar anaesthesia in cataract surgery. METHODS: Ninety patients were allocated randomly to receive 8 ml of a mixture of equal parts of bupivacaine 0.75% and lidocaine 2% or an equal volume of L-bupivacaine and lidocaine 2%. Hyaluronidase 15 IU ml(-1) was added to both solutions. RESULTS: There were significant differences between the groups in clinical end-points. The median time at which the block was adequate to start surgery was 4 min (interquartile range 4-8 min) in the bupivacaine group and 8 min (5-12 min) in the L-bupivacaine group (P=0.002). Median ocular and eyelid movement scores were similarly significantly decreased in the bupivacaine group compared with the L-bupivacaine group at all times (PAla) abolishes partial agonist activity of steroidal antiandrogens. AB - Mutation of a single amino acid in the ligand-binding domain (LBD) of the human androgen receptor (hAR) can induce functional abnormalities in androgen binding, stabilization of active conformation, or interaction with coactivators. The Gly708Ala and Gly708Val substitutions are associated with partial and complete androgen insensitivity syndromes, respectively. In this work, we introduced Ala, Val, and aromatic Phe mutations at position 708 on helix H3 of the hAR-LBD and tested the functional and structural consequences on hAR activity in the presence of steroidal or nonsteroidal agonists and antagonists. The residues involved in the specific recognition of these androgen ligands were identified and analyzed in the light of in vitro biological experiments and the 3D hAR-LBD structure. Our study demonstrated that the Gly708Ala mutation influenced the agonist versus antagonist activity of the ligands and confirmed the crucial role of this residue within the ligand-binding pocket (LBP) in the modulation of androgen agonists. The Gly708Ala mutation transformed the antiandrogen cyproterone acetate (CPA), a partial agonist, into a pure antiandrogen, and the pure nonsteroidal antiandrogen hydroxyflutamide in a partial agonist. From the docking studies, we suggest that CPA acts on AR through the novel mechanism called "passive antagonism". PMID- 12644580 TI - Brominated derivatives of noscapine are potent microtubule-interfering agents that perturb mitosis and inhibit cell proliferation. AB - Noscapine, a microtubule-interfering agent, has been shown to arrest mitosis, to induce apoptosis, and to have potent antitumor activity. We report herein that two brominated derivatives of noscapine, 5-bromonoscapine (5-Br-nosc) and reduced 5-bromonoscapine (Rd 5-Br-nosc), have higher tubulin binding activity than noscapine and affect tubulin polymerization differently from noscapine. In addition, they are able to arrest cell cycle progression at mitosis at concentrations much lower than noscapine. Interestingly, whereas noscapine arrested cells have nearly normal bipolar spindles, cells arrested by 5-Br-nosc and Rd 5-Br-nosc form multipolar spindles. Nevertheless, noscapine and the two derivatives all affect the attachment of chromosomes to spindle microtubules and they impair the tension across paired kinetochores to similar degrees. 5-Br-nosc and Rd 5-Br-nosc are also more active than noscapine in inhibiting the proliferation of various human cancer cells, including those that are resistant to paclitaxel and epothilone. Our results thus indicate a great potential for the use of 5-Br-nosc and Rd 5-Br-nosc both as biological tools for studying microtubule-mediated processes and as chemotherapeutic agents for the treatment of human cancers. PMID- 12644581 TI - Involvements of voltage-independent Ca2+ channels and phosphoinositide 3-kinase in endothelin-1-induced PYK2 tyrosine phosphorylation. AB - We demonstrated recently that endothelin-1 (ET-1) activates two types of Ca(2+) permeable nonselective cation channels [designated nonselective cation channel (NSCC)-1 and NSCC-2] and a store-operated Ca(2+) channel (SOCC) in rabbit internal carotid artery vascular smooth muscle cells (ICA VSMCs). These channels can be distinguished by their sensitivity to Ca(2+) channel blockers 1-(beta-[3 (4-methoxyphenyl) propoxy]-4-methoxyphenethyl)-1H-imidazole hydrochloride (SK&F 96365) and (R,S)-(3,4-dihydro-6,7-dimethoxy-isochinolin-1-yl)-2-phenyl-N,N-di[2 (2,3,4-trimethoxyphenyl)ethyl]acetamid mesylate (LOE 908). NSCC-1 is sensitive to LOE 908 and resistant to SK&F 96365, NSCC-2 is sensitive to both LOE 908 and SK&F 96365, and SOCC is resistant to LOE 908 and sensitive to SK&F 96365. The purpose of the present study was to identify the Ca(2+) channels involved in the ET-1 induced, proline-rich tyrosine kinase 2 (PYK2) phosphorylation in ICA VSMCs. Based on sensitivity to nifedipine, an L-type voltage-operated Ca(2+) channel (VOCC) blocker, Ca(2+) influx through VOCC seems to play a minor role in the ET-1 induced PYK2 phosphorylation. In the presence of nifedipine, PYK2 phosphorylation was abolished by blocking Ca(2+) influx through NSCC-1, NSCC-2, and SOCC. The phosphoinositide 3-kinase (PI3K) inhibitors wortmannin and 2-(4-morpholinyl)-8 phenyl-4H-1-benzopyran-4-one (LY 294002), inhibited ET-1-induced Ca(2+) influx through NSCC-2 and SOCC. In addition, these inhibitors blocked PYK2 phosphorylation that depends on Ca(2+) influx through NSCC-2 and SOCC. These results indicate that 1) Ca(2+) influx through NSCC-1, NSCC-2, and SOCC plays essential roles in ET-1-induced PYK2 phosphorylation, 2) NSCC-2 and SOCC are stimulated by ET-1 via a PI3K-dependent cascade, whereas NSCC-1 is stimulated via a PI3K-independent cascade, and 3) PI3K is involved in the PYK2 phosphorylation that depends on Ca(2+) influx through SOCC and NSCC-2. PMID- 12644582 TI - A Leishmania major nucleobase transporter responsible for allopurinol uptake is a functional homolog of the Trypanosoma brucei H2 transporter. AB - Nucleobase transporters play an important role in the physiology of protozoan parasites, because these organisms are purine auxotrophs and rely entirely on salvage of these vital compounds. Purine transporters have also been shown to mediate the uptake of important antiparasitic drugs. In the current study, we investigated the uptake of [(3)H]adenine, [(3)H]hypoxanthine, and [(3)H]allopurinol, an antileishmanial hypoxanthine analog, by Leishmania major. These compounds were all taken up by a single high-affinity transporter, LmNBT1, with K(m) values of 4.6 +/- 0.9, 0.71 +/- 0.07, and 54 +/- 3 microM, respectively. Guanine and xanthine fully inhibited [(3)H]adenine transport, with K(i) values of 2.8 +/- 0.7 and 23 +/- 8 microM. Using purine analogs, an inhibitor profile for LmNBT1 was obtained, which allowed the construction of a quantitative model for the interactions between the transporter binding site and the permeant. The model predicts that hypoxanthine was bound through hydrogen bonds to N(1)H, N3, N7, and N(9)H of the purine ring, with a total Gibbs free energy of -39.5 kJ/mol. The interactions with adenine were similar, except for a weak hydrogen bond to N1 (unprotonated in adenine). The predicted mode of substrate binding for LmNBT1 was almost identical to that for the Trypanosoma brucei H2 (TbH2) transporter. It is proposed that the architecture of their respective binding sites is very similar and that LmNBT1 can be named a functional homolog of TbH2. PMID- 12644583 TI - Unexpected induction of the human connexin 43 promoter by the ras signaling pathway is mediated by a novel putative promoter sequence. AB - Connexin 43 (Cx43) is essential for survival and is tightly regulated at the transcriptional and post-transcriptional levels. A number of previous studies have demonstrated altered expression in malignant tissues, and in the presence of carcinogenic factors. We examined the effect of protooncogenes of Cx43 expression, and found no effect on Cx43 promoter activity in cells transformed with Src or erbB2. On the other hand, we identified and characterized a novel sequence that mediates Cx43 promoter regulation in cell lines engineered to overexpress H-Ras. Compared with wild-type NIH3T3 cells, both Cx43 mRNA and protein levels are increased in NIH3T3-Ras cells. The H-Ras+ cells also have enhanced Cx43 promoter activation, which is inhibited by the MEK1 inhibitor 2' amino-3'-methoxyflavone (PD98059), suggesting that Ras-mediated Cx43 overexpression is via the mitogen activated protein kinase kinase/extracellular signal-regulated pathway. Deletion analysis of the Cx43 promoter revealed a 200 bp region downstream of the Cx43 transcription start site as the minimal sequence essential for the Ras-mediated Cx43 up-regulation. Using this 200-base pair fragment in electrophoretic mobility shift assays, we identified one main protein complex that binds efficiently and is more abundant in nuclear extracts from NIH3T3-Ras and MCF7-Ras cells compared with their matched controls. This complex selectively recognizes a consensus sequence, AGTTCAATCA, located at positions +149 to +158 of the Cx43 promoter. Supershift assays identified the 90-kDa heat shock protein (HSP90) and c-Myc as constituents of this DNA-binding complex. Treatment of cells with the HSP90 inhibitor geldanamycin resulted in repression of the Cx43 promoter activity, and inhibits binding of the complex to the Cx43 promoter. Coimmunoprecipitation studies confirmed the interaction between endogenous HSP90 and c-Myc. This study provides evidence that the transcriptional up-regulation of Cx43 by Ras-Raf-MAPK is mediated via the interaction of a novel Cx43 promoter element with a protein complex that contains both HSP90 and c-Myc. PMID- 12644584 TI - Mechanism of action of Gq to inhibit G beta gamma modulation of CaV2.2 calcium channels: probed by the use of receptor-G alpha tandems. AB - The stable interaction of a G-protein coupled receptor and a particular partner G protein was made possible by creating tandems between the alpha(2A) adrenergic receptor (alpha(2A)-R) and pertussis toxin-resistant mutants of different G alpha subunits of heterotrimeric G-proteins. Both alpha(2A)-R-G alpha(o) and alpha(2A) R-G alpha(i) proved able to reconstitute agonist-induced voltage-dependent inhibition of N-type calcium channels (Ca(V)2.2) similar to the wild-type alpha(2A)-R when expressed in COS-7 cells. The interaction of G(q) with the G(i/o) signaling pathways was studied by expressing either G alpha(q) or a chimeric construct based on G alpha(q) containing the last five amino acids of G alpha(z), which is activated by alpha(2A)-R. It was found that G alpha(qz5) activated by the wild-type alpha(2A)-R inhibited Ca(V)2.2 currents in a voltage independent fashion. Furthermore, G alpha(qz5) counteracted the voltage-dependent inhibition resulting from alpha(2A)-R-G alpha(o) activation. We subsequently investigated the basis for the behavior of G alpha(qz5). Our evidence suggests that this occurs as a result of a downstream effect of activation of G alpha(qz5) because it was blocked by C-terminal construct of phospholipase C beta 1. Furthermore it is likely to occur in part via protein kinase C (PKC) activation, because the PKC activator phorbol dibutyrate mimicked the effects of G alpha(qz5) in alpha(2A)-R-G alpha(o)-transfected cells. Conversely, cells expressing both alpha(2A)-R-G alpha(o) and G alpha(qz5) exhibited a partial restoration of voltage-dependent inhibition in the presence of the PKC inhibitor bisindolylmaleimide I (GF 109203X). The potential sites of phosphorylation are discussed. PMID- 12644585 TI - Involvement of organic cation transporter 1 in the lactic acidosis caused by metformin. AB - Biguanides are a class of drugs widely used as oral antihyperglycemic agents for the treatment of type 2 diabetes mellitus, but they are associated with lactic acidosis, a lethal side effect. We reported previously that biguanides are good substrates of rat organic cation transporter 1 (Oct1; Slc22a1) and, using Oct1(-/ ) mice, that mouse Oct1 is responsible for the hepatic uptake of a biguanide, metformin. In the present study, we investigated whether the liver is the key organ for the lactic acidosis. When mice were given metformin, the blood lactate concentration significantly increased in the wild-type mice, whereas only a slight increase was observed in Oct1(-/-) mice. The plasma concentration of metformin exhibited similar time profiles between the wild-type and Oct1(-/-) mice, suggesting that the liver is the key organ responsible for the lactic acidosis. Furthermore, the extent of the increase in blood lactate caused by three different biguanides (metformin, buformin, and phenformin) was compared with the abilities to reduce oxygen consumption in isolated rat hepatocytes. When rats were given each of these biguanides, the lactate concentration increased significantly. This effect was dose-dependent, and the EC(50) values of metformin, buformin, and phenformin were 734, 119, and 4.97 microM, respectively. All of these biguanides reduced the oxygen consumption by isolated rat hepatocytes in a concentration-dependent manner. When the concentration required to reduce the oxygen consumption to 75% of the control value (from 0.40 to 0.29 micromol/min/mg protein) was compared with the EC(50) value obtained in vivo, a clear correlation was observed among the three biguanides, suggesting that oxygen consumption in isolated rat hepatocytes can be used as an index of the incidence of lactic acidosis. PMID- 12644586 TI - Anthracyclines induce accumulation of iron in ferritin in myocardial and neoplastic cells: inhibition of the ferritin iron mobilization pathway. AB - Anthracyclines are potent antitumor agents that cause cardiotoxicity at high cumulative doses. Because anthracycline cardiotoxicity is attributed to their ability to avidly bind iron (Fe), we examined the effect of anthracyclines on intracellular Fe trafficking in neoplastic cells and differentiated cardiomyocytes. In both cell types, incubation with doxorubicin (DOX) resulted in a significant (p < 0.004) accumulation of Fe in the storage protein, ferritin. Pulse-chase experiments using control cells demonstrated that within 6 h, the majority of (59)Fe donated from transferrin was incorporated into ferritin. Over longer incubation periods up to 18 to 24 h, (59)Fe was subsequently mobilized from ferritin into other compartments in control cells. However, anthracyclines inhibited ferritin-(59)Fe redistribution during the 18- to 24-h period, resulting in a significant (p < 0.0003) 3- to 5-fold accumulation of ferritin-(59)Fe compared with control cells. The increase in ferritin-(59)Fe after a 24-h incubation with DOX could not be correlated with increased ferritin expression, suggesting that (59)Fe accumulation occurred in pre-existing ferritin. In addition to DOX, other redox-cycling agents (i.e., menadione and paraquat) also increased ferritin-(59)Fe levels. Moreover, the intracellular superoxide scavenger, Mn(III) tetrakis(4-benzoic acid)-porphyrin complex, partially prevented the ability of DOX and menadione at inducing this effect. Hence, superoxide generation by these compounds could play a role in causing ferritin (59)Fe accumulation. This study is the first to demonstrate the effect of anthracyclines at inhibiting Fe mobilization from ferritin, resulting in marked Fe accumulation within the molecule. This response may have consequences in terms of the cytotoxic effects of anthracyclines. PMID- 12644587 TI - Gemcitabine potentiates cisplatin cytotoxicity and inhibits repair of cisplatin DNA damage in ovarian cancer cell lines. AB - Synergistic cytotoxicity between cisplatin and the nucleoside analog gemcitabine was observed in a panel of cisplatin-sensitive (2008, A2780) and -resistant (2008/C13*5.25, A2780/CP70) human ovarian cell lines. Previous studies have suggested a role for DNA repair in the mechanism of synergy between the two drugs. We therefore further investigated the hypothesis that the synergistic cytotoxicity between gemcitabine and cisplatin in these cell lines may be caused by gemcitabine-mediated inhibition of cisplatin intrastrand adduct (IA) and interstand cross-link (ICL) repair. The effect of gemcitabine on the accumulation and repair of cisplatin IA and ICL in each cell line was then measured directly using gene-specific quantitative polymerase chain reaction and denaturation/renaturation techniques, respectively. Pretreatment of 2008 cells with 1 microM gemcitabine for 2 h before exposure to cisplatin for 7 h enhanced the accumulation of cisplatin IA and ICL by 50 and 40%, respectively (P < 0.05), above that induced by cisplatin alone. To investigate the possibility that the increased accumulation of cisplatin lesions was caused by inhibition of removal of cisplatin damage, 2008 cells were incubated with 200 microM cisplatin for 5 h in the presence and absence of gemcitabine and then a further 8 h in the absence of cisplatin. Only 57% IA were removed in the combination treated cells compared with 74% in cisplatin control cells. Similarly, repair of cisplatin ICL was inhibited in the gemcitabine-treated cells compared with the cells treated with cisplatin only (60 versus 72%). These findings demonstrate a direct inhibitory effect of gemcitabine on the repair of cisplatin IA and ICL and suggest a mechanistic basis for the cytotoxic synergy between the two drugs. PMID- 12644588 TI - Characterization of celecoxib and valdecoxib binding to cyclooxygenase. AB - Two compounds (celecoxib and valdecoxib) from the diarylheterocycle class of cyclooxygenase inhibitors were radiolabeled and used to characterize their binding to cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), several single point variants of COX-2 (Val523Ile, Tyr355Ala, Arg120Ala, Arg120Gln, Arg120Asn) and one triple-point variant of COX-2 [Val523Ile, Arg513His, Val434Ile (IHI)]. We demonstrate highly specific and saturable binding of these inhibitors to COX-2. Under the same assay conditions, little or no specific binding to COX-1 could be detected. The affinity of [(3)H]celecoxib for COX-2 (K(D) = 2.3 nM) was similar to the affinity of [(3)H]valdecoxib (K(D) = 3.2 nM). The binding to COX-2 seems to be both rapid and slowly reversible with association rates of 5.8 x 10(6)/M/min and 4.5 x 10(6)/M/min and dissociation rates of 14 x 10(-3)/min (t(1/2) = 50 min) and 7.0 x 10(-3)/min (t(1/2) = 98 min) for [(3)H]celecoxib and [(3)H]valdecoxib, respectively. These association rates increased (4- to 11-fold) when the charged arginine residue located at the entrance to the main hydrophobic channel was mutated to smaller uncharged amino acids (Arg120Ala, Arg120Gln, and Arg120Asn). Mutation of residues located within the active site of COX-2 that define a 'side pocket' (Tyr355Ala, Val523Ile, IHI) of the main channel had a greater effect on the dissociation rate than the association rate. These mutations, which modified the shape of and access to the 'side pocket', affected the binding affinity of [(3)H]valdecoxib more than that of [(3)H]celecoxib. These binding studies provide direct insight into the properties and binding constants of celecoxib and valdecoxib to COX-2. PMID- 12644589 TI - P2Y receptors modulate ion channel function through interactions involving the C terminal domain. AB - Nucleotide stimulation of G(q)-coupled P2Y receptors expressed in Xenopus laevis oocytes produces the activation of an endogenous voltage-gated ion channel, previously identified as the transient inward (T(in)) channel. Expression of human P2Y(1), human P2Y(2), rat P2Y(6), human P2Y(11), or skate P2Y receptors in oocytes resulted in modulation of the voltage dependence and inactivation gating of the channel. Expression of the human P2Y(4) receptor, rat M(1)-muscarinic receptor, and human B(1)-bradykinin receptor did not alter the properties of the T(in) channel. Replacement of the C-terminal domain of the human B(1)-bradykinin receptor with the C-terminal domains of either the human P2Y(1) or human P2Y(2) receptor resulted in voltage dependence and inactivation-gating properties, respectively, of the T(in) channel that were similar to those elicited by the respective native P2Y receptor. Systematic truncation of the C-terminal region of the human P2Y(1) receptor identified a short region responsible for modulation of the T(in) channel. This region contains a conserved sequence motif found in all P2Y receptors that modulates the voltage dependence of the T(in) channel. Synthetic 20-mer peptides from the C-terminal domains of human P2Y(1) and P2Y(2) receptors produced a shift in the voltage dependence and slowed inactivation gating, respectively, after injection into oocytes expressing human B(1) bradykinin or truncated human P2Y(1) receptors. These results indicate that certain P2Y receptors are capable of modulating the voltage sensitivity and inactivation gating of an endogenous oocyte ion channel through interactions involving the C-terminal region of the receptor. Such modulation of ion channel function could also exist in native mammalian cells that express P2Y receptors. PMID- 12644590 TI - Activation of endothelial nitric-oxide synthase by tumor necrosis factor-alpha: a novel pathway involving sequential activation of neutral sphingomyelinase, phosphatidylinositol-3' kinase, and Akt. AB - Activation of endothelial nitric-oxide synthase (eNOS) has been shown to occur through various pathways involving increases in the cytosolic Ca(2+) concentration, activation of the phosphatidylinositol-3' kinase/Akt pathway, as well as regulation by other kinases and by protein-protein interactions. We have recently reported that eNOS, expressed in an inducible HeLa Tet-off cell line, is activated by tumor necrosis factor-alpha (TNF-alpha) in a previously undescribed pathway that involves the lipid messenger ceramide. We have now characterized this pathway. We report here that eNOS activation in response to TNF-alpha correlated with phosphorylation of Akt at Ser 473 and of eNOS itself at Ser 1179. Akt and eNOS phosphorylation, as well as eNOS activation, were blocked by inhibitors of both phosphatidylinositol-3' kinase and neutral sphingomyelinase. In contrast, although acid sphingomyelinase was also stimulated by TNF-alpha, its inhibition was without effect. The activation of neutral sphingomyelinase triggered by TNF-alpha was insensitive to phosphatidylinositol-3' kinase inhibitors. Taken together, these results indicate that eNOS activation by TNF alpha occurs through sequential activation of neutral sphingomyelinase and of the phosphatidylinositol-3' kinase/Akt pathway. The time course of eNOS activation induced through this pathway was markedly different from that triggered by ATP and epidermal growth factor, which activate eNOS through an increase in intracellular Ca(2+) concentration and through a sphingomyelinase-independent stimulation of the phosphatidylinositol-3' kinase/Akt pathway, respectively. The novel pathway of activation of eNOS described here may have broad biological relevance because neutral sphingomyelinase is activated not only by TNF-alpha but also by a variety of other physiological and pathological stimuli. PMID- 12644591 TI - Gamma-hydroxybutyric acid and diazepam antagonize a rapid increase in GABA(A) receptors alpha(4) subunit mRNA abundance induced by ethanol withdrawal in cerebellar granule cells. AB - Both benzodiazepines and gamma-hydroxybutyric acid (GHB) are used to treat alcohol withdrawal syndrome. The molecular basis for this therapeutic efficacy was investigated with primary cultures of rat cerebellar granule cells. Long-term exposure of these cells to ethanol (100 mM, 5 days) reduced the abundance of mRNAs encoding the gamma(2)L and gamma(2)S subunits of the GABA type A receptor ( 32 and -23%, respectively) but failed to affect that of alpha(1), alpha(4), or alpha(6) subunit mRNAs. Subsequent ethanol withdrawal resulted in decreases in the amounts of alpha(1) (-29%), alpha(6) (-27%), gamma(2)L (-64%), and gamma(2)S (-76%),subunit mRNAs that were maximal after 6 to 12 h. In contrast, 3 h after ethanol withdrawal, the abundance of the alpha(4) subunit mRNA was increased by 46%. Ethanol withdrawal did not affect neuronal morphology but reduced cellular metabolic activity. The increase in alpha(4) subunit was confirmed by functional studies showing a positive action of flumazenil in patch clamp recordings of GABA stimulated currents after ethanol withdrawal. Diazepam (10 microM) or GHB (100 mM) prevented the increase in the amount of the alpha(4) subunit mRNA, the metabolic impairment, and the positive action of flumazenil induced by ethanol withdrawal but failed to restore the expression of the alpha(1) and gamma(2) subunits. The antagonism by GHB seems not to be mediated by a direct action at GABA(A)R because GHB failed to potentiate the effects of GABA or diazepam on Cl( ) currents mediated by GABA type A receptor. PMID- 12644593 TI - Species-specific transcriptional activity of synthetic flavonoids in guinea pig and mouse cells as a result of differential activation of the aryl hydrocarbon receptor to interact with dioxin-responsive elements. AB - To investigate possible species-specificity of aryl hydrocarbon receptor (AhR) mediated signal transduction pathways, activities of 2,3,7,8-tetrochlorodibenzo-p dioxin (TCDD) and six synthetic flavonoids were evaluated in mouse hepatoma and guinea pig adenocarcinoma cells transfected with an AhR-responsive luciferase reporter. Rank order potency in these two cell lines was similar for the ability of these flavonoids to antagonize TCDD-induced reporter gene expression. However, in the presence of flavone alone, a species-specific difference in agonist activity was observed. In guinea pig cells, several flavonoids demonstrated agonist activity up to 50% of the maximum TCDD response. In mouse cells, however, no significant agonist activity was observed at the same concentrations based on luciferase enzyme activity, protein expression, and mRNA analysis. Moreover, competitive ligand-binding assays, using [(3)H]TCDD in cytosolic fractions, demonstrated that 3'-methoxy-4'-nitroflavone had a similar IC(50) in both recombinant cell lines, suggesting that the flavone has similar binding affinity to receptors from both species. However, electrophoretic mobility shift assay using the cytosolic fractions demonstrated that this flavone elicited binding to the DRE by guinea pig but not mouse AhR complex. The dependence of the AhR in this differential interaction was further demonstrated using in vitro synthesized guinea pig and mouse Ah receptors and mouse Arnt. Together, these data suggest that the differential agonist/antagonist activity of these flavone derivatives is caused by the efficacy of these flavonoids in eliciting an AhR conformation that recognizes regulatory response elements in a species-specific manner. PMID- 12644592 TI - The cannabinoid CB1 receptor antagonist SR141716 increases Acrp30 mRNA expression in adipose tissue of obese fa/fa rats and in cultured adipocyte cells. AB - This study investigates the effects of SR141716, a selective CB(1) receptor antagonist that reduces food intake and body weight of rodents, on Acrp30 mRNA expression in adipose tissue. Acrp30, a plasma protein exclusively expressed and secreted by adipose tissue, has been shown to induce free fatty acid oxidation, hyperglycemia and hyperinsulinemia decrease, and body weight reduction. We report that N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H pyrazole-3-carboximide hydrochloride (SR141716) treatment once daily (10 mg/kg/d, i.p.) from 2 to 14 days reduced body weight and stimulated Acrp30 mRNA expression in adipose tissue of obese Zucker (fa/fa) rats. In parallel, the hyperinsulinemia associated with this animal model was reduced by SR141716 treatment. In cultured mouse adipocytes (3T3 F442A), SR141716 (25 to 100 nM) also induced an overexpression of Acrp30 mRNA and protein. In addition, in adipose tissue of CB(1)-receptor knockout mice, SR141716 had no effect on Acrp30 mRNA expression, demonstrating a CB(1) receptor mediating effect. Furthermore, RT-PCR analysis revealed that rat adipose tissue and 3T3 F442A adipocytes expressed CB(1) receptor mRNA. Relative quantification of this expression revealed an up regulation (3- to 4-fold) of CB(1) receptor mRNA expression in adipose tissue of obese (fa/fa) rats and in differentiated 3T3 F442A adipocytes compared with lean rats and undifferentiated adipocytes, respectively. Western blot analysis revealed the presence of CB(1) receptors in 3T3 F442A adipocytes, and their expression was up-regulated in differentiated cells. These results show that SR141716 stimulated Acrp30 mRNA expression in adipose tissue by an effect on adipocytes, and reduced hyperinsulinemia in obese (fa/fa) rats. These hormonal regulations may participate in the body weight reduction induced by SR141716 and suggest a role of metabolic regulation in the antiobesity effect of SR141716. PMID- 12644594 TI - Resveratrol increases serine15-phosphorylated but transcriptionally impaired p53 and induces a reversible DNA replication block in serum-activated vascular smooth muscle cells. AB - Resveratrol (RV), a polyphenolic stilbene derivative, has been proposed to exert a plethora of beneficial cardiovascular effects. Of these, in particular, inhibition of vascular smooth muscle cell (VSMC) proliferation shows great promise for preventing cardiovascular disease. In the present study, we show that RV leads to a reversible arrest in early S phase of the VSMC cycle, accompanied by an accumulation of hyperphosphorylated retinoblastoma protein. In contrast to studies with other cell systems, RV decreases cellular levels of the cyclin dependent kinase inhibitors p21(Cip1) and p27(Kip1). This is of particular interest because phosphorylated p53 protein (serine(15)) is strongly enhanced by this substance. We further found that RV only slightly inhibits phosphorylation of Erk 1/2, protein kinase B/Akt, and p70(S6) kinase upon serum stimulation. Thus, inhibition of these kinases is not likely to contribute to the cell cycle effect of RV. Importantly, the observed S phase arrest is not linked to an increase in apoptotic cell death: there was no detectable increase in apoptotic nuclei and in levels of the proapoptotic protein Bax. This is the first study elucidating the molecular pathways mediating the antiproliferative properties of RV in VSMCs. PMID- 12644596 TI - Opposite regulation of the human paraoxonase-1 gene PON-1 by fenofibrate and statins. AB - The human paraoxonase-1 (PON-1) is a serum high-density lipoprotein-associated phosphotriesterase secreted mainly by the liver. This enzyme is able to hydrolyze toxic organophosphate xenobiotics, endogenous oxidized phospholipids, and homocysteine thiolactone. Physiologically, it is thought to protect against cardiovascular diseases. The level of PON-1 gene expression is a major determinant of paraoxonase-1 status but little is known regarding the regulation of this gene. We identified several transcription start sites and characterized the regulation of its promoter by fibrates and statins. In HuH7 human hepatoma cells, the PON-1 secreted enzymatic activity and mRNA levels were increased by fenofibric acid (approximately 70%) and decreased by several statins (approximately 50%). Transient and stable transfection assays in HuH7 cells indicated that the modulation of the mRNA and enzymatic activity levels could be accounted for by the regulation of the PON-1 gene promoter activity by these drugs. These effects are probably not mediated by the PPAR alpha because over expression of this receptor decreased the fibrate effect and did not modify statins activity. The repressive effect of statins is reversed by mevalonate and 22(R)-hydroxycholesterol, suggesting the involvement of the liver X receptor in the mechanism. The opposite effects of fenofibrate and statins could be consistent with clinical data on homocysteine levels after hypolipidemic drug treatment. Regarding the toxicological aspects, the induction achieved with fenofibric acid, although limited, could increase organophosphate metabolism and may be relevant in certain conditions for protective treatments. PMID- 12644595 TI - Antitumor and cellular pharmacological properties of a novel platinum(IV) complex: trans-[PtCl(2)(OH)(2)(dimethylamine) (isopropylamine)]. AB - The antitumor and cellular pharmacological properties of the trans-Pt(IV) complex, trans-[PtCl(2)(OH)(2)(dimethylamine)(isopropylamine)] (compound 2) has been evaluated in comparison with its corresponding trans-Pt(II) counterpart, trans-[PtCl(2)(dimethylamine)(isopropylamine)] (compound 1). The results reported here indicate that compound 2 markedly circumvents cisplatin resistance in 41McisR and CH1cisR ovarian tumor cell lines endowed with different mechanisms of resistance (decreased platinum accumulation and enhanced DNA repair/tolerance, respectively). However, compound 1 is able to circumvent cisplatin resistance only in CH1cisR cells. Interestingly, at equitoxic concentrations, compounds 1 and 2 induce a higher amount of apoptotic cells than cisplatin in CH1cisR cells. Moreover, the number of apoptotic cells induced by compounds 1 and 2 correlates with their ability to form DNA interstrand cross-links in CH1cisR cells. Although compounds 1 and 2 showed remarkable cytotoxic activity, only compound 2 was able to inhibit the growth of CH1 human ovarian carcinoma xenografts in mice. Binding studies with serum albumin indicate that compound 1 possesses a much higher reactivity against albumin than compound 2. Moreover, the level of binding of compound 1 to plasma proteins during the period 15 min to 1 h after administration to mice (15 mg/kg, i.p.) is 2.5-fold higher than that of compound 2. Therefore, the lack of in vivo antitumor activity shown by compound 1 might be related to its extracellular inactivation before reaching the tumor site because of its high rate of binding to plasma proteins. PMID- 12644597 TI - Genetic component of sensorimotor capacity in rats. AB - Creation of generalized genetic models for low and high sensorimotor capacity would be important tools for resolution of this complex trait. As proof-of principle we estimated phenotypic variation and narrow-sense heritability (h(2)) of sensorimotor capacity in 19 families of genetically heterogeneous N:NIH rats and in 11 strains of inbred rats. Sensorimotor capacity was defined as the time a rat remained on an accelerating rotorod. N:NIH rats recorded variation in rotorod scores that ranged from 3- to 7-fold. The value of h(2), estimated from offspring parent regression across one generation, was 0.68 for females and 0.74 for males in N:NIH rats. In inbred rats, h(2) was estimated by partitioning phenotypic variation into additive genetic and environmental components and averaged 0.39 in females and 0.48 in males. These results demonstrate a heritable component to sensorimotor capacity sufficient for success in developing contrasting genetic models by divergent artificial selection in rats. PMID- 12644600 TI - Measurement of the profile angle and the aesthetic analysis of the facial profile. AB - Cephalometric measurement of the face in terms of aesthetics can be difficult and misleading due to the variability of the intra-cranial reference lines. Extra cranial references are more accurate, but can be time-consuming to apply. The Aesthetic Horizontal is an intuitive datum line related to the 'photographic position' of the head, which is expedient in use and clinically relevant. A new and straightforward technique is presented for transferring the Aesthetic Horizontal directly from the patient to any recent radiograph, which can then be used as the reference line for an aesthetic analysis of the facial profile. The instrument used for measuring the profile angle and the transfer is readily constructed from a protractor and small weight. The technique can also be used to transfer any other orientation (e.g. Natural Head Position or Natural Head Posture) from the patient to a recent radiograph. PMID- 12644598 TI - Identification of endothelial cell genes by combined database mining and microarray analysis. AB - Vascular endothelial cells maintain the interface between the systemic circulation and soft tissues and mediate critical processes such as inflammation in a vascular bed-selective fashion. To expand our understanding of the genetic pathways that underlie these specific functions, we have focused on the identification of novel genes that are differentially expressed in all endothelial cells, as well as restricted groups of this cell type. Virtual subtraction was conducted employing gene expression data deposited in public databases and 384 genes identified. These genes were spotted on custom microarrays, along with 288 genes identified through subtraction cloning from TGF beta-stimulated endothelial cells. Arrays were evaluated with RNA samples representing endothelial cells cultured from four vascular sources and five non endothelial cell types. These studies identified 64 pan-endothelial markers that were differentially expressed with at least a threefold difference (range 3- to 55-fold). In addition, differences in gene expression profiles among endothelial cells from different vascular beds were identified. Validation of these findings was performed by RNA blot expression studies, and a number of the novel genes were shown to be expressed under angiogenic conditions in the developing mouse embryo. The combined tools of database mining and transcriptional profiling thus provide expanded knowledge of endothelial cell gene expression and endothelial cell biology. PMID- 12644601 TI - Broken orthodontic trans-palatal archwire stuck to the throat of orthodontic patient: is it strange? AB - Trans-palatal arch wire is in common use in orthodontic practice. This case report of broken trans-palatal wire, swallowed and stuck to the throat of an orthodontic patient, is presented in the hope that it will stress the importance of careful examination of the appliance at each scheduled visit. PMID- 12644602 TI - Idiopathic gingival hyperplasia and orthodontic treatment: a case report. AB - There are many reasons for gingival hyperplasia. Mostly, proper oral hygiene is sufficient to achieve normal healthy gingiva. In some situations, however, gingival hyperplasia is drug-induced or can be a manifestation of a genetic disorder. In the latter, it may exist as an isolated abnormality or as part of a syndrome. If orthodontic treatment is needed in patients with gingival hyperplasia, both orthodontic and periodontal aspects need to be considered. Extreme hereditary gingival fibromatosis was periodontally treated, by removal of all gingival excess using flaps and gingivectomies. After a follow-up period, the orthodontic treatment started with fixed appliances. Monthly periodontal check ups (scaling and polishing) were scheduled to control the gingival inflammation. After the orthodontic treatment, permanent retention was applied, once more followed by a complete gingivectomy in both maxilla and mandible. One of the most important keys to successful treatment of hyperplasia patients is the cooperation between the periodontist and the orthodontist. PMID- 12644604 TI - Clinical trials in orthodontics I: demographic details of clinical trials published in three orthodontic journals between 1989 and 1998. AB - AIM: To test the hypothesis that there is insufficient evidence available, from clinical trials, to allow evidence-based decisions to be made on the effectiveness of orthodontic treatment. OBJECTIVES: To identify reports of orthodontic clinical trials and assess their demographic characteristics. DESIGN: A retrospective, observational study. SETTING: The American Journal of Orthodontics and Dentofacial Orthopedics, British Journal of Orthodontics, and European Journal Orthodontics. DATA SOURCE: Clinical trials published between 1989 and 1998. METHOD: A hand-search was performed to identify all clinical trials. The journal and year of publication, research method, interventions, and sample size of the trials reported were recorded. RESULTS: One-hundred-and-fifty five trial reports were identified of which 56 (36.1%) were published from 1989 to 1993 and 99 (69%) from 1994 to 1998. Ninety-nine (69%) reports were published in the AJO-DO, 18 (11.6%) in the BJO and 38 (24.5%) in the EJO. Eighty-five (54.8%) were reports of randomized controlled trials and 70 (45.2%) of controlled clinical trials. The interventions most frequently assessed were bonding materials (21.9%), growth modification treatments (21.3%), and oral hygiene procedures (9.0%). The median sample size was 32 (IQR 19.5, 50). CONCLUSION: There is sufficient evidence available from clinical trials to warrant doing systematic reviews of orthodontic clinical trials to aid decision-making. PMID- 12644605 TI - Inactivated periods of constant orthodontic forces related to desirable tooth movement in rats. AB - AIM: To examine the effects of inactive periods of force on the amount of tooth displacement and root resorption during experimental tooth movement in rats. SAMPLE: Sixty 11-week-old male Sprague-Dawley rats. METHOD: The maxillary first molar (M1) was moved mesially using a removable titanium-nickel alloy closed coil spring for 14 days. The rats were divided into four groups with, 0, 1, 4, and 9 hours of inactivation per day. RESULTS: Tooth displacement in the 0- and 1-hour groups was significantly greater than that in the 9-hour group. The area of root resorption in the 4- and 9-hour groups was significantly less than that in the 0- and 1-hour groups. There was no significant difference in root resorption between 0- and 1-hour groups, and also between 4- and 9-hour groups. CONCLUSION: The distance of tooth displacement gradually decreased as the inactive period increased, whereas root resorption suddenly decreased between 1 and 4 hours of inactive orthodontic force. PMID- 12644606 TI - Orthodontic treatment need and self-perception of 11-16-year-old Saudi Arabian children with a sensory impairment attending special schools. AB - AIM: To determine the self-perception and need for orthodontic treatment in young sensory [visual (VI) and hearing (HI)] impaired children attending special schools in Riyadh, Saudi Arabia. Also, to determine if gender and social class background influence the rating and self-perception of malocclusion among the children. LOCATION: Riyadh, Saudi Arabia. DESIGN: A prospective study on orthodontic treatment need in sensory impaired children. SUBJECTS: Seventy-seven VI, 210 HI, and 494 control (C) children aged 11-16 years. METHOD: The aesthetic component (AC) of the Index of Orthodontic Treatment Need (IOTN) was determined using the standard 10 pictures for the C group and HI with a modified version (tactile graphic) for VI. The dental health component (DHC) and AC of IOTN were used to allocate each child to no need, borderline need and definite need for treatment subgroups. RESULTS: Sixty-five per cent of VI, 21.8 per cent HI, and 18.7 per cent of the C were perceived to be in need of orthodontic treatment. However, 55.8 VI, 43 per cent HI and 34 per cent C were rated for treatment need based upon the AC. The difference between the examiner and the child's rating of treatment need was found to be statistically significant among the HI and control children (P < 0.001). CONCLUSION: The VI children who were scored for treatment by the examiner as having need for treatment had similar perceptions of their treatment need irrespective of their social background. Male VI children had a higher DHC score, but both VI and HI males had a higher normative and self perceived need based on AC. PMID- 12644607 TI - Does articulating study casts make a difference to treatment planning? AB - OBJECTIVE: To assess whether articulating casts in centric relation (CR) compared with intercuspal position (ICP) makes a difference to treatment planning. DESIGN: Reliability analysis. SUBJECTS: Ten orthodontists. METHODS: Twenty case vignettes were examined on three occasions: twice with the casts in ICP and once in CR. A series of dichotomous decisions were made relating to the treatment need and treatment mechanics. MAIN OUTCOME MEASURES: The changes in treatment decisions were examined. Intra-examiner agreement between the two hand-held cast assessments (H1 v. H2) and between the first set of hand-held casts compared with the articulated casts (H1 v. A1) were evaluated using the kappa statistic. The differences between the kappa statistics for H1 v. H2 and H1 v. A1 were then tested with the Wilcoxon matched pairs signed rank sum test. RESULTS: The only statistically significant change in the kappa score between H1 v. H2 and H1 v. A1 was for the extraction decision (P= 0.007). No other statistically significant differences were found for the other treatment decisions, although trends were identified for orthognathic surgery and anchorage support decisions. CONCLUSION: Routine articulation of study models for all orthodontic patients is not supported by the results of this study. Articulation of the study models did not affect the treatment planning decisions in a meaningful manner. Further work with selected samples is required to determine if articulation is helpful for specific malocclusions. PMID- 12644608 TI - An ex vivo investigation to compare orthodontic bonding using a 4-META-based adhesive or a composite adhesive to acid-etched and sandblasted enamel. AB - OBJECTIVE: This study investigates the shear-peel orthodontic bond strengths of brackets bonded with an unfilled acrylic resin containing 4-META (MCP Bond or a no-mix composite adhesive (Right On) to acid-etched or sandblasted enamel. DESIGN: Ex vivo. MATERIALS AND METHODS: Eighty human pre-molar teeth were separated into four equal groups, according to the adhesive used and method of enamel pre-treatment. Group I-Right On with enamel etched using phosphoric acid for 30 seconds. Group II-Right On with enamel sandblasted using 50- microm alumina particles at 80 psi for 3 seconds. Group III-MCP Bond with enamel etched using phosphoric acid for 30 seconds. Group IV-MCP Bond with enamel sandblasted using 50- microm alumina particles at 80 psi for 3 seconds. Subsequently, the specimens were stored in distilled water for 24 hours prior to bond strength testing using an Instron universal testing machine. Each debonded tooth was scored using the adhesive remnant index (ARI) to determine the site of bond failure. RESULTS: The mean bond strength (1 SD) were Group I: 10.7 (2.7) MPa, Group II: 5.3 (1.3) MPa, Group III: 15.9 (3.4) MPa, Group IV: 15.0 (2.2) MPa. Statistical analysis using one-way analysis of variance and Tukey test found no statistical difference between Group III and Group IV (P > 0.05), but the other groups were statistically different from each other (P < 0.05). The data were found to fit the Weibull distribution and Weibull analysis showed stress required for a 5 per cent probability of failure was: Group I: 5.77 MPa; Group II: 3.32 MPa; Group III: 10.31 MPa; Group IV: 10.58 MPa. Chi-square test showed a statistically significant difference existed between the ARI scores (P < 0.001), principally through less adhesive remnants being observed on the sandblasted specimens. CONCLUSION: The adhesive containing 4-META achieved significantly higher bond strengths than the composite adhesive, particularly in the case of sandblasted enamel. PMID- 12644609 TI - European Board of Orthodontists: a professional challenge. AB - The background to the formation of the European Board is given and the necessary procedures for obtaining certification of the European Board of Orthodontists (EBO) are described. An example case report is included to give the reader an indication of the type of detail required for each and every case presented. Recommendations are given for prospective candidates who might consider attempting EBO certification in the future. PMID- 12644610 TI - How to.... find the evidence. PMID- 12644613 TI - Current products and practice section: religious, cultural, and ethical dilemmas in orthodontics. AB - There are potential religious, cultural and ethical dilemmas facing patients who are undergoing orthodontic treatment. Opinions were collected from religious and cultural leaders, as well as from non-religious groups who take an ethical standpoint on certain medical or dental treatment. Factors relating to ingredients in orthodontic products, timing of orthodontic appointments, and the effects of religious fasting are all discussed. It is important to recognize all patients' opinions and beliefs when planning and undertaking orthodontic treatment. PMID- 12644614 TI - Orthodontic speciality training in the UK. PMID- 12644615 TI - Orthodontic speciality training in the UK. PMID- 12644616 TI - Lack of training numbers restricting the number of training places available. PMID- 12644619 TI - Fish or chips? AB - Cell membranes are not simply barriers separating intracellular from extracellular space. Rather, they represent a dynamic high-turnover system that adapts to current demands. During inflammation, prostaglandins and leukotrienes are formed from membrane-derived phospholipids. Encouraging improvements in critically ill patients were observed after nutritional replacement of long-chain omega-6 fatty acids with long-chain omega-3-fatty acids, contained in fish oil. PMID- 12644618 TI - "The force be with you": ATP in gut mechanosensory transduction. AB - Nucleotides such as ATP and UTP are everywhere. They play a key role in transducing mechanosensory signals via P2Y receptors in the large intestine or colon, leading to secretion of the sensory mediator 5-HT, and act as autocrine, paracrine, or neurocrine mediators in neural reflexes regulating chloride secretion. PMID- 12644620 TI - Alveolar flooding at high altitude: failure of reabsorption? AB - Na-driven fluid reabsorption prevents alveolar fluid accumulation. Hypoxia augments fluid filtration by hemodynamic effects and inhibits Na reabsorption in cultured cells. A combination of both effects in vivo causes alveolar flooding, edema, thickening of the diffusion barrier for oxygen, and hypoxemia, a life threatening situation for mountaineers in remote high-altitude mountain ranges. PMID- 12644621 TI - Annexin 1: an endogenous anti-inflammatory protein. AB - A hallmark of inflammation is the mobilization of blood-borne leukocytes across microvessels to kill and remove the invading pathogen. For its damaging potential, leukocyte movement is finely regulated, and endogenous pathways exist to ensure the time dependency of this process. Annexin 1 and its receptor(s) are one example of these pathways. PMID- 12644622 TI - Angiogenesis and vascular remodeling by intussusception: from form to function. AB - During most instances of angiogenesis, not only are the capillaries or terminal vessels generated and modified, but the supplying vascular system is subjected to remodeling as well. Intussusception, i.e., transluminal pillar formation, is one essential mechanism for growth, arborization, bifurcation remodeling, and pruning. Complex and efficient vascular beds can thus be generated by local interactions between vascular cells and hemodynamic conditions. PMID- 12644623 TI - Self glycosphingolipids: new antigens recognized by autoreactive T lymphocytes. AB - T cells may recognize glycolipids and lipids of bacterial and self origin associated with the CD1 antigen-presenting molecules. Understanding the mechanisms governing CD1-self glycolipid interaction will provide information on the molecular rules of glycolipid presentation and suggest new approaches to immunotherapy. PMID- 12644625 TI - Arteriovenous pairing: a determinant of capillary exchange. AB - Venuloarteriolar signaling helps mediate microvascular function and dysfunction. Mediators produced at venular sites of inflammation appear to constrict arterioles and increase capillary permeability. In contrast, venules beneficially dilate arterioles to enhance capillary flow according to metabolic demand. These mechanisms are altered with cardiovascular risk factors, contributing to microvascular complications. PMID- 12644624 TI - Hypoxia, hormones, and red blood cell function in chick embryos. AB - The red blood cell function of avian embryos is regulated by cAMP. Adenosine A(2A) and beta-adrenergic receptor activation during hypoxic conditions cause changes in the hemoglobin oxygen affinity and CO(2) transport. Furthermore, experimental evidence suggests a general involvement of cAMP in terminal differentiation of avian erythroblasts. PMID- 12644628 TI - A haplotype map of the human genome. PMID- 12644629 TI - The proof of the pudding is in the eating: editorial focus on "Hyperphagia, not hypometabolism, causes early onset obesity in melanocortin-4-receptor knockout mice". PMID- 12644630 TI - Super models. AB - Model organisms have been used over a century to understand basic, conserved biological processes. The study of these experimental systems began with genetics and development, moved into molecular and cellular biology, and most recently propelled into functional genomics and proteomics. The goal of this review is simple: to discuss the place of model organisms in "The Age of the Ome": the genome, the transcriptome, and the proteome. This review will address the following questions. What exactly is a model organism? What characteristics make an excellent model system? Using the yeast Saccharomyces cerevisiae and the nematode Caenorhabditis elegans as examples, this review will discuss these issues with the aim of demonstrating how model organisms remain indispensable scientific tools for understanding complex biological pathways and human disease. PMID- 12644631 TI - Proteomic approach to coronary atherosclerosis shows ferritin light chain as a significant marker: evidence consistent with iron hypothesis in atherosclerosis. AB - Coronary artery disease (CAD) is the leading cause of mortality and morbidity in developed nations. We hypothesized that CAD is associated with distinct patterns of protein expression in the coronary arteries, and we have begun to employ proteomics to identify differentially expressed proteins in diseased coronary arteries. Two-dimensional (2-D) gel electrophoresis of proteins and subsequent mass spectrometric analysis identified the ferritin light chain as differentially expressed between 10 coronary arteries from patients with CAD and 7 coronary arteries from normal individuals. Western blot analysis indicated significantly increased expression of the ferritin light chain in the diseased coronary arteries (1.41 vs. 0.75; P = 0.01). Quantitative real-time PCR analysis showed that expression of ferritin light chain mRNA was decreased in diseased tissues (0.70 vs. 1.17; P = 0.013), suggesting that increased expression of ferritin light chain in CAD coronary arteries may be related to increased protein stability or upregulation of expression at the posttranscriptional level in the diseased tissues. Ferritin light chain protein mediates storage of iron in cells. We speculate that increased expression of the ferritin light chain may contribute to pathogenesis of CAD by modulating oxidation of lipids within the vessel wall through the generation of reactive oxygen species. Our results provide in situ proteomic evidence consistent with the "iron hypothesis," which proposes an association between excessive iron storage and a high risk of CAD. However, it is also possible that the increased ferritin expression in diseased coronary arteries is a consequence, rather than a cause, of CAD. PMID- 12644632 TI - Hyperphagia, not hypometabolism, causes early onset obesity in melanocortin-4 receptor knockout mice. AB - Previous studies on mice with melanocortin-4 receptor gene (MC4r) knockout have focused on obese adults. Because humans with functional MC4r mutations show early onset obesity, we determined the onset of excessive fat deposition in 10- to 56 day-old mice, taking into account sex and litter influences. Total body fat content of MC4r-/- on day 35 and MC4r+/- on day 56 significantly exceeds that of MC4r+/+. Plasma leptin levels increase in proportion to fat mass. According to cumulative food intake and energy expenditure measurements from day 21 to 35, onset of excessive fat deposition in MC4r-/- is fueled by hyperphagia and counteracted partially by hypermetabolism. In 35- to 56-day-old mice, arcuate nucleus neuropeptide Y (NPY) mRNA decreases and pro-opiomelanocortin (POMC) mRNA increases with fat content and plasma leptin levels independently of genotype. Taking into account fat content by ANCOVA reveals, however, increases in both NPY mRNA and POMC mRNA due to melanocortin-4 receptor (MC4R) deficiency. We conclude that hyperphagia, not hypometabolism, is the primary disturbance initiating excessive fat deposition in MC4R-deficient mice at weaning and that the overall changes in NPY and POMC expression tend to antagonize the onset of excessive fat deposition. PMID- 12644633 TI - Gene expression profiling of Caco-2 BBe cells suggests a role for specific signaling pathways during intestinal differentiation. AB - We examined the pattern of gene expression resulting from spontaneous differentiation of Caco-2 BBe cells to gain insight into the molecular changes necessary for enterocyte differentiation. RNA was prepared from cells harvested at three cell stages: proliferating (50% confluent, 2 days in culture), postproliferative nondifferentiated (8 days), and differentiated (15 days). Gene expression profiles were determined using Affymetrix Human Genome U95A GeneChips. Differentially expressed genes were identified following statistical analysis (i.e., ANOVA, bootstrapping adjustments to P values, false detection rate criterion). We identified 1,150 unique genes as differentially expressed; expression of 48.6% fell and 46% increased from 2 to 15 days, while 5.4% had expression that either peaked or dipped at 8 days. Genes expressed during differentiation included several small-intestine-specific genes involved in nutrient transport/metabolism, e.g., DCT1, hephaestin, folate receptor 1, sucrase isomaltase, and apolipoproteins CI, CIII, B100, H, and M, indicating that this colonic adenocarcinoma cell line has a hybrid colonocyte/enterocyte phenotype. Patterns of gene expression based upon functional classification suggest a role for cell-cell/cell-matrix interactions, suppression of Wnt signaling, and activation of TGFbeta and phosphatidylinositol 3-kinase pathways during enterocyte differentiation. PMID- 12644636 TI - The hot and the classic. PMID- 12644634 TI - Novel mechanisms of T-cell and dendritic cell activation revealed by profiling of psoriasis on the 63,100-element oligonucleotide array. AB - A global picture of gene expression in the common immune-mediated skin disease, psoriasis, was obtained by interrogating the full set of Affymetrix GeneChips with psoriatic and control skin samples. We identified 1,338 genes with potential roles in psoriasis pathogenesis/maintenance and revealed many perturbed biological processes. A novel method for identifying transcription factor binding sites was also developed and applied to this dataset. Many of the identified sites are known to be involved in immune response and proliferation. An in-depth study of immune system genes revealed the presence of many regulating cytokines and chemokines within involved skin, and markers of dendritic cell (DC) activation in uninvolved skin. The combination of many CCR7+ T cells, DCs, and regulating chemokines in psoriatic lesions, together with the detection of DC activation markers in nonlesional skin, strongly suggests that the spatial organization of T cells and DCs could sustain chronic T-cell activation and persistence within focal skin regions. PMID- 12644637 TI - Summaries of legume genomics projects from around the globe. Community resources for crops and models. PMID- 12644638 TI - A TILLING reverse genetics tool and a web-accessible collection of mutants of the legume Lotus japonicus. PMID- 12644639 TI - Legumes: importance and constraints to greater use. PMID- 12644640 TI - Legume natural products: understanding and manipulating complex pathways for human and animal health. PMID- 12644641 TI - Can we improve the nutritional quality of legume seeds? PMID- 12644642 TI - Recent advances in legume transformation. PMID- 12644643 TI - The rest of the iceberg. Legume diversity and evolution in a phylogenetic context. PMID- 12644644 TI - Floral development in legumes. PMID- 12644645 TI - Axillary meristem development. Budding relationships between networks controlling flowering, branching, and photoperiod responsiveness. PMID- 12644646 TI - Symbiosis, inventiveness by recruitment? PMID- 12644647 TI - A cluster of genes encodes the two types of chalcone isomerase involved in the biosynthesis of general flavonoids and legume-specific 5-deoxy(iso)flavonoids in Lotus japonicus. AB - Leguminous plants produce 5-deoxyflavonoids and 5-deoxyisoflavonoids that play essential roles in legume-microbe interactions. Together with chalcone polyketide reductase and cytochrome P450 2-hydroxyisoflavanone synthase, the chalcone isomerase (CHI) of leguminous plants is fundamental in the construction of these ecophysiologically active flavonoids. Although CHIs of nonleguminous plants isomerize only 6'-hydroxychalcone to 5-hydroxyflavanone (CHIs with this function are referred to as type I), leguminous CHIs convert both 6'-deoxychalcone and 6' hydroxychalcone to 5-deoxyflavanone and 5-hydroxyflavanone, respectively (referred to as type II). In this study, we isolated multiple CHI cDNAs (cCHI1 cCHI3) from a model legume, Lotus japonicus. In contrast to previous observations, the amino acid sequence of CHI2 was highly homologous to nonleguminous CHIs, whereas CHI1 and CHI3 were the conventional leguminous type. Furthermore, genome sequence analysis revealed that four CHI genes (CHI1-3 and a putative gene, CHI4) form a tandem cluster within 15 kb. Biochemical analysis with recombinant CHIs expressed in Escherichia coli confirmed that CHI1 and CHI3 are type II CHIs and that CHI2 is a type I CHI. The occurrence of both types of CHIs is probably common in leguminous plants, and it was suggested that type II CHIs evolved from an ancestral CHI by gene duplication and began to produce 5 deoxy(iso)flavonoids along with the establishment of the Fabaceae. PMID- 12644648 TI - A diffusible factor from arbuscular mycorrhizal fungi induces symbiosis-specific MtENOD11 expression in roots of Medicago truncatula. AB - Using dual cultures of arbuscular mycorrhizal (AM) fungi and Medicago truncatula separated by a physical barrier, we demonstrate that hyphae from germinating spores produce a diffusible factor that is perceived by roots in the absence of direct physical contact. This AM factor elicits expression of the Nod factor inducible gene MtENOD11, visualized using a pMtENOD11-gusA reporter. Transgene induction occurs primarily in the root cortex, with expression stretching from the zone of root hair emergence to the region of mature root hairs. All AM fungi tested (Gigaspora rosea, Gigaspora gigantea, Gigaspora margarita, and Glomus intraradices) elicit a similar response, whereas pathogenic fungi such as Phythophthora medicaginis, Phoma medicaginis var pinodella and Fusarium solani f.sp. phaseoli do not, suggesting that the observed root response is specific to AM fungi. Finally, pMtENOD11-gusA induction in response to the diffusible AM fungal factor is also observed with all three M. truncatula Nod(-)/Myc(-) mutants (dmi1, dmi2, and dmi3), whereas the same mutants are blocked in their response to Nod factor. This positive response of the Nod(-)/Myc(-) mutants to the diffusible AM fungal factor and the different cellular localization of pMtENOD11-gusA expression in response to Nod factor versus AM factor suggest that signal transduction occurs via different pathways and that expression of MtENOD11 is differently regulated by the two diffusible factors. PMID- 12644650 TI - Nod factor elicits two separable calcium responses in Medicago truncatula root hair cells. AB - Modulation of intracellular calcium levels plays a key role in the transduction of many biological signals. Here, we characterize early calcium responses of wild type and mutant Medicago truncatula plants to nodulation factors produced by the bacterial symbiont Sinorhizobium meliloti using a dual-dye ratiometric imaging technique. When presented with 1 nM Nod factor, root hair cells exhibited only the previously described calcium spiking response initiating 10 min after application. Nod factor (10 nM) elicited an immediate increase in calcium levels that was temporally earlier and spatially distinct from calcium spikes occurring later in the same cell. Nod factor analogs that were structurally related, applied at 10 nM, failed to initiate this calcium flux response. Cells induced to spike with low Nod factor concentrations show a calcium flux response when Nod factor is raised from 1 to 10 nM. Plant mutants previously shown to be deficient for the calcium spiking response (dmi1 and dmi2) exhibited an immediate, truncated calcium flux with 10 nM Nod factor, demonstrating a competence to respond to Nod factor but an impaired ability to generate a full biphasic response. These results demonstrate that the legume root hair cell exhibits two independent calcium responses to Nod factor triggered at different agonist concentrations and suggests an early branch point in the Nod factor signal transduction pathway. PMID- 12644649 TI - Conformation of a group 2 late embryogenesis abundant protein from soybean. Evidence of poly (L-proline)-type II structure. AB - Late embryogenesis abundant (LEA) proteins are members of a large group of hydrophilic, glycine-rich proteins found in plants, algae, fungi, and bacteria known collectively as hydrophilins that are preferentially expressed in response to dehydration or hyperosmotic stress. Group 2 LEA (dehydrins or responsive to abscisic acid) proteins are postulated to stabilize macromolecules against damage by freezing, dehydration, ionic, or osmotic stress. However, the structural and physicochemical properties of group 2 LEA proteins that account for such functions remain unknown. We have analyzed the structural properties of a recombinant form of a soybean (Glycine max) group 2 LEA (rGmDHN1). Differential scanning calorimetry of purified rGmDHN1 demonstrated that the protein does not display a cooperative unfolding transition upon heating. Ultraviolet absorption and circular dichroism spectroscopy revealed that the protein is in a largely hydrated and unstructured conformation in solution. However, ultraviolet absorption and circular dichroism measurements collected at different temperatures showed that the protein exists in equilibrium between two extended conformational states: unordered and left-handed extended helical or poly (L proline)-type II structures. It is estimated that 27% of the residues of rGmDHN1 adopt or poly (L-proline)-type II-like helical conformation at 12 degrees C. The content of extended helix gradually decreases to 15% as the temperature is increased to 80 degrees C. Studies of the conformation of the protein in solution in the presence of liposomes, trifluoroethanol, and sodium dodecyl sulfate indicated that rGmDHN1 has a very low intrinsic ability to adopt alpha-helical structure and to interact with phospholipid bilayers through amphipathic alpha helices. The ability of the protein to remain in a highly extended conformation at low temperatures could constitute the basis of the functional role of GmDHN1 in the prevention of freezing, desiccation, ionic, or osmotic stress-related damage to macromolecular structures. PMID- 12644651 TI - Expression of an expansin gene is correlated with root elongation in soybean. AB - Expansin is a family of proteins that catalyze long-term expansion of cell walls and has been considered a principal protein that affects cell expansion in plants. We have identified the first root-specific expansin gene in soybean (Glycine max), GmEXP1, which may be responsible for root elongation. Expression levels of GmEXP1 were very high in the roots of 1- to 5-d-old seedlings, in which rapid root elongation takes place. Furthermore, GmEXP1 mRNA was most abundant in the root tip region, where cell elongation occurs, but scarce in the region of maturation, where cell elongation ceases, implying that its expression is closely related to root development processes. In situ hybridization showed that GmEXP1 transcripts were preferentially present in the epidermal cells and underlying cell layers in the root tip of the primary and secondary roots. Ectopic expression of GmEXP1 accelerated the root growth of transgenic tobacco (Nicotiana tabacum) seedlings, and the roots showed insensitivity to obstacle-touching stress. These results imply that the GmEXP1 gene plays an important role in root development in soybean, especially in the elongation and/or initiation of the primary and secondary roots. PMID- 12644652 TI - Dual genetic pathways controlling nodule number in Medicago truncatula. AB - We report the isolation and characterization of a new Medicago truncatula hyper nodulation mutant, designated sunn (super numeric nodules). Similar to the previously described ethylene-insensitive mutant sickle, sunn exhibits a 10-fold increase in the number of nodules within the primary nodulation zone. Despite this general similarity, these two mutants are readily distinguished based on anatomical, genetic, physiological, and molecular criteria. In contrast to sickle, where insensitivity to ethylene is thought to be causal to the hyper nodulation phenotype (R.V. Penmetsa, D.R. Cook [1997] Science 275: 527-530), nodulation in sunn is normally sensitive to ethylene. Nevertheless, sunn exhibits seedling root growth that is insensitive to ethylene, although other aspects of the ethylene triple response are normal; these observations suggest that hormonal responses might condition the sunn phenotype in a manner distinct from sickle. The two mutants also differ in the anatomy of the nodulation zone: Successful infection and nodule development in sunn occur predominantly opposite xylem poles, similar to wild type. In sickle, however, both infection and nodulation occur randomly throughout the circumference of the developing root. Genetic analysis indicates that sunn and sickle correspond to separate and unlinked loci, whereas the sunn/skl double mutant exhibits a novel and additive super-nodulation phenotype. Taken together, these results suggest a working hypothesis wherein sunn and sickle define distinct genetic pathways, with skl regulating the number and distribution of successful infection events, and sunn regulating nodule organogenesis. PMID- 12644653 TI - The Sym35 gene required for root nodule development in pea is an ortholog of Nin from Lotus japonicus. AB - Comparative phenotypic analysis of pea (Pisum sativum) sym35 mutants and Lotus japonicus nin mutants suggested a similar function for the PsSym35 and LjNin genes in early stages of root nodule formation. Both the pea and L. japonicus mutants are non-nodulating but normal in their arbuscular mycorrhizal association. Both are characterized by excessive root hair curling in response to the bacterial microsymbiont, lack of infection thread initiation, and absence of cortical cell divisions. To investigate the molecular basis for the similarity, we cloned and sequenced the PsNin gene, taking advantage of sequence information from the previously cloned LjNin gene. An RFLP analysis on recombinant inbred lines mapped PsNin to the same chromosome arm as the PsSym35 locus and direct evidence demonstrating that PsNin is the PsSym35 gene was subsequently obtained by cosegregation analysis and sequencing of three independent Pssym35 mutant alleles. L. japonicus and pea root nodules develop through different organogenic pathways, so it was of interest to compare the expression of the two orthologous genes during nodule formation. Overall, a similar developmental regulation of the PsNin and LjNin genes was shown by the transcriptional activation in root nodules of L. japonicus and pea. In the indeterminate pea nodules, PsNin is highly expressed in the meristematic cells of zone I and in the cells of infection zone II, corroborating expression of LjNin in determinate nodule primordia. At the protein level, seven domains, including the putative DNA binding/dimerization RWP RK motif and the PB1 heterodimerization domain, are conserved between the LjNIN and PsNIN proteins. PMID- 12644654 TI - Syntenic relationships between Medicago truncatula and Arabidopsis reveal extensive divergence of genome organization. AB - Arabidopsis and Medicago truncatula represent sister clades within the dicot subclass Rosidae. We used genetic map-based and bacterial artificial chromosome sequence-based approaches to estimate the level of synteny between the genomes of these model plant species. Mapping of 82 tentative orthologous gene pairs reveals a lack of extended macrosynteny between the two genomes, although marker collinearity is frequently observed over small genetic intervals. Divergence estimates based on non-synonymous nucleotide substitutions suggest that a majority of the genes under analysis have experienced duplication in Arabidopsis subsequent to divergence of the two genomes, potentially confounding synteny analysis. Moreover, in cases of localized synteny, genetically linked loci in M. truncatula often share multiple points of synteny with Arabidopsis; this latter observation is consistent with the large number of segmental duplications that compose the Arabidopsis genome. More detailed analysis, based on complete sequencing and annotation of three M. truncatula bacterial artificial chromosome contigs suggests that the two genomes are related by networks of microsynteny that are often highly degenerate. In some cases, the erosion of microsynteny could be ascribed to the selective gene loss from duplicated loci, whereas in other cases, it is due to the absence of close homologs of M. truncatula genes in Arabidopsis. PMID- 12644655 TI - Identification and characterization of nodulation-signaling pathway 2, a gene of Medicago truncatula involved in Nod actor signaling. AB - Bacterially derived Nod factor is critical in the establishment of the legume/rhizobia symbiosis. Understanding the mechanisms of Nod factor perception and signal transduction in the plant will greatly advance our understanding of this complex interaction. Here, we describe the identification of a new locus, nodulation-signaling pathway 2 (NSP2), of Medicago truncatula that is involved in Nod factor signaling. Mutants at this locus are blocked for Nod factor-induced gene expression and show a reduced root hair deformation response. nsp2 plants also show a complete absence of infection and cortical cell division following Sinorhizobium meliloti inoculation. Nod factor-induced calcium spiking, one of the earliest responses tested, is still functional in these mutant plants. We conclude that the gene NSP2 is a component of the Nod factor signal transduction pathway that lies downstream of the calcium-spiking response. PMID- 12644656 TI - Dual intracellular localization and targeting of aminoimidazole ribonucleotide synthetase in cowpea. AB - De novo purine biosynthesis is localized to both mitochondria and plastids isolated from Bradyrhizobium sp.-infected cells of cowpea (Vigna unguiculata L. Walp) nodules, but several of the pathway enzymes, including aminoimidazole ribonucleotide synthetase (AIRS [EC 6.3.3.1], encoded by Vupur5), are encoded by single genes. Immunolocalization confirmed the presence of AIRS protein in both organelles. Enzymatically active AIRS was purified separately from nodule mitochondria and plastids. N-terminal sequencing showed that these two isoforms matched the Vupur5 cDNA sequence but were processed at different sites following import; the mitochondrial isoform was five amino acids longer than the plastid isoform. Electrospray tandem mass spectrometry of a trypsin digest of mitochondrial AIRS identified two internal peptides identical with the amino acid sequence deduced from Vupur5 cDNA. Western blots of proteins from mitochondria and plastids isolated from root tips showed a single AIRS protein present at low levels in both organelles. (35)S-AIRS protein translated from a Vupur5 cDNA was imported into isolated pea (Pisum sativum) leaf chloroplasts in vitro by an ATP dependent process but not into import-competent mitochondria from several plant and non-plant sources. Components of the mature protein are likely to be important for import because the N-terminal targeting sequence was unable to target green fluorescent protein to either chloroplasts or mitochondria in Arabidopsis leaves. The data confirm localization of the protein translated from the AIRS gene in cowpea to both plastids and mitochondria and that it is cotargeted to both organelles, but the mechanism underlying import into mitochondria has features that are yet to be identified. PMID- 12644657 TI - A phylogenomic investigation of CYCLOIDEA-like TCP genes in the Leguminosae. AB - Numerous TCP genes (transcription factors with a TCP domain) occur in legumes. Genes of this class in Arabidopsis (TCP1) and snapdragon (Antirrhinum majus; CYCLOIDEA) have been shown to be asymmetrically expressed in developing floral primordia, and in snapdragon, they are required for floral zygomorphy (bilaterally symmetrical flowers). These genes are therefore particularly interesting in Leguminosae, a family that is thought to have evolved zygomorphy independently from other zygomorphic angiosperm lineages. Using a phylogenomic approach, we show that homologs of TCP1/CYCLOIDEA occur in legumes and may be divided into two main classes (LEGCYC group I and II), apparently the result of an early duplication, and each class is characterized by a typical amino acid signature in the TCP domain. Furthermore, group I genes in legumes may be divided into two subclasses (LEGCYC IA and IB), apparently the result of a duplication near the base of the papilionoid legumes or below. Most papilionoid legumes investigated have all three genes present (LEGCYC IA, IB, and II), inviting further work to investigate possible functional difference between the three types. However, within these three major gene groups, the precise relationships of the paralogs between species are difficult to determine probably because of a complex history of duplication and loss with lineage sorting or heterotachy (within-site rate variation) due to functional differentiation. The results illustrate both the potential and the difficulties of orthology determination in variable gene families, on which the phylogenomic approach to formulating hypotheses of function depends. PMID- 12644658 TI - crinkle, a novel symbiotic mutant that affects the infection thread growth and alters the root hair, trichome, and seed development in Lotus japonicus. AB - To elucidate the mechanisms involved in Rhizobium-legume symbiosis, we examined a novel symbiotic mutant, crinkle (Ljsym79), from the model legume Lotus japonicus. On nitrogen-starved medium, crinkle mutants inoculated with the symbiont bacterium Mesorhizobium loti MAFF 303099 showed severe nitrogen deficiency symptoms. This mutant was characterized by the production of many bumps and small, white, uninfected nodule-like structures. Few nodules were pale-pink and irregularly shaped with nitrogen-fixing bacteroids and expressing leghemoglobin mRNA. Morphological analysis of infected roots showed that nodulation in crinkle mutants is blocked at the stage of the infection process. Confocal microscopy and histological examination of crinkle nodules revealed that infection threads were arrested upon penetrating the epidermal cells. Starch accumulation in uninfected cells and undeveloped vascular bundles were also noted in crinkle nodules. Results suggest that the Crinkle gene controls the infection process that is crucial during the early stage of nodule organogenesis. Aside from the symbiotic phenotypes, crinkle mutants also developed morphological alterations, such as crinkly or wavy trichomes, short seedpods with aborted embryos, and swollen root hairs. crinkle is therefore required for symbiotic nodule development and for other aspects of plant development. PMID- 12644659 TI - Nylon filter arrays reveal differential gene expression in proteoid roots of white lupin in response to phosphorus deficiency. AB - White lupin (Lupinus albus) adapts to phosphorus deficiency (-P) by the development of short, densely clustered lateral roots called proteoid (or cluster) roots. In an effort to better understand the molecular events mediating these adaptive responses, we have isolated and sequenced 2,102 expressed sequence tags (ESTs) from cDNA libraries prepared with RNA isolated at different stages of proteoid root development. Determination of overlapping regions revealed 322 contigs (redundant copy transcripts) and 1,126 singletons (single-copy transcripts) that compile to a total of 1,448 unique genes (unigenes). Nylon filter arrays with these 2,102 ESTs from proteoid roots were performed to evaluate global aspects of gene expression in response to -P stress. ESTs differentially expressed in P-deficient proteoid roots compared with +P and -P normal roots include genes involved in carbon metabolism, secondary metabolism, P scavenging and remobilization, plant hormone metabolism, and signal transduction. PMID- 12644660 TI - Proteome analysis. Novel proteins identified at the peribacteroid membrane from Lotus japonicus root nodules. AB - The peribacteroid membrane (PBM) forms the structural and functional interface between the legume plant and the rhizobia. The model legume Lotus japonicus was chosen to study the proteins present at the PBM by proteome analysis. PBM was purified from root nodules by an aqueous polymer two-phase system. Extracted proteins were subjected to a global trypsin digest. The peptides were separated by nanoscale liquid chromatography and analyzed by tandem mass spectrometry. Searching the nonredundant protein database and the green plant expressed sequence tag database using the tandem mass spectrometry data identified approximately 94 proteins, a number far exceeding the number of proteins reported for the PBM hitherto. In particular, a number of membrane proteins like transporters for sugars and sulfate; endomembrane-associated proteins such as GTP binding proteins and vesicle receptors; and proteins involved in signaling, for example, receptor kinases, calmodulin, 14-3-3 proteins, and pathogen response related proteins, including a so-called HIR protein, were detected. Several ATPases and aquaporins were present, indicating a more complex situation than previously thought. In addition, the unexpected presence of a number of proteins known to be located in other compartments was observed. Two characteristic protein complexes obtained from native gel electrophoresis of total PBM proteins were also analyzed. Together, the results identified specific proteins at the PBM involved in important physiological processes and localized proteins known from nodule-specific expressed sequence tag databases to the PBM. PMID- 12644661 TI - The Medicago species A2-type cyclin is auxin regulated and involved in meristem formation but dispensable for endoreduplication-associated developmental programs. AB - Phytohormones as well as temporal and spatial regulation of the cell cycle play a key role in plant development. Here, we investigated the function and regulation of an alfalfa (Medicago sativa) A2-type cyclin in three distinct root developmental programs: in primary and secondary root development, nodule development, and nematode-elicited gall formation. Using transgenic plants carrying the Medsa;cycA2;2 promoter-beta-glucuronidase gene fusion, in combination with other techniques, cycA2;2 expression was localized in meristems and proliferating cells in the lateral root and nodule primordia. Rapid induction of cycA2;2 by Nod factors demonstrated that this gene is implicated in cell cycle activation of differentiated cells developing to nodule primordia. Surprisingly, cycA2;2 was repressed in the endoreduplicating, division-arrested cells both during nodule development and formation of giant cells in nematode-induced galls, indicating that CycA2;2 was dispensable for S-phase in endoreduplication cycles. Overexpression of cycA2;2 in transgenic plants corresponded to wild type protein levels and had no apparent phenotype. In contrast, antisense expression of cycA2;2 halted regeneration of somatic embryos, suggesting a role for CycA2;2 in the formation or activity of apical meristems. Expression of cycA2;2 was up regulated by auxins, as expected from the presence of auxin response elements in the promoter. Moreover, auxin also affected the spatial expression pattern of this cyclin by shifting the cycA2;2 expression from the phloem to the xylem poles. PMID- 12644662 TI - Mapping the proteome of barrel medic (Medicago truncatula). AB - A survey of six organ-/tissue-specific proteomes of the model legume barrel medic (Medicago truncatula) was performed. Two-dimensional polyacrylamide gel electrophoresis reference maps of protein extracts from leaves, stems, roots, flowers, seed pods, and cell suspension cultures were obtained. Five hundred fifty-one proteins were excised and 304 proteins identified using peptide mass fingerprinting and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Nanoscale high-performance liquid chromatography coupled with tandem quadrupole time-of-flight mass spectrometry was used to validate marginal matrix-assisted laser desorption ionization time-of-flight mass spectrometry protein identifications. This dataset represents one of the most comprehensive plant proteome projects to date and provides a basis for future proteome comparison of genetic mutants, biotically and abiotically challenged plants, and/or environmentally challenged plants. Technical details concerning peptide mass fingerprinting, database queries, and protein identification success rates in the absence of a sequenced genome are reported and discussed. A summary of the identified proteins and their putative functions are presented. The tissue specific expression of proteins and the levels of identified proteins are compared with their related transcript abundance as quantified through EST counting. It is estimated that approximately 50% of the proteins appear to be correlated with their corresponding mRNA levels. PMID- 12644664 TI - Pollination-, development-, and auxin-specific regulation of gibberellin 3beta hydroxylase gene expression in pea fruit and seeds. AB - To understand further how pollination, seeds, auxin (4-chloroindole-3-acetic acid [4-Cl-IAA]), and gibberellins (GAs) regulate GA biosynthesis in pea (Pisum sativum) fruit, we studied expression of the gene PsGA3ox1 that codes for the enzyme that converts GA(20) to biologically active GA(1) using real-time reverse transcription-polymerase chain reaction analysis. PsGA3ox1 mRNA levels were minimally detectable in prepollinated pericarps and ovules (-2 d after anthesis [DAA]), increased dramatically after pollination (0 DAA), then decreased by 1 DAA. Seed PsGA3ox1 mRNA levels increased at 4 DAA and again 8 to 12 DAA, when seed development was rapid. Pericarp PsGA3ox1 mRNA levels peaked coincidentally with rapid pod diameter expansion (6-10 DAA) to accommodate the growing seeds. The effects of seeds and hormones on the expression of pericarp PsGA3ox1 were investigated over a 24-h treatment period. Pericarp PsGA3ox1 mRNA levels gradually increased from 2 to 3 DAA when seeds were present; however, when the seeds were removed, the pericarp transcript levels dramatically declined. When 2 DAA deseeded pericarps were treated with 4-Cl-IAA, PsGA3ox1 mRNA levels peaked 4 h after hormone treatment (270-fold increase), then decreased. PsGA3ox1 mRNA levels in deseeded pericarps treated with indole-3-acetic acid or GA(3) were the same or lower than deseeded controls. These data show that PsGA3ox1 is expressed and developmentally regulated in pea pericarps and seeds. These data also show that pericarp PsGA3ox1 expression is hormonally regulated and suggest that the conversion of GA(20) to GA(1) occurs in the pericarp and is regulated by the presence of seeds and 4-Cl-IAA for fruit growth. PMID- 12644663 TI - Expression of the apyrase-like APY1 genes in roots of Medicago truncatula is induced rapidly and transiently by stress and not by Sinorhizobium meliloti or Nod factors. AB - The model legume Medicago truncatula contains at least six apyrase-like genes, five of which (MtAPY1;1, MtAPY1;2, MtAPY1;3, MtAPY1;4, and MtAPY1;5) are members of a legume-specific family, whereas a single gene (MtAPY2) has closer homologs in Arabidopsis. Phylogenetic analysis has revealed that the proteins encoded by these two plant gene families are more similar to yeast (Saccharomyces cerevisiae) GDA1 and to two proteins encoded by newly described mammalian genes (ENP5 and 6) than they are to mammalian CD39- and CD39-like proteins. Northern analyses and analyses of the frequencies of expressed sequence tags (ESTs) in different cDNA libraries suggest that in roots, leaves, and flowers, the more highly expressed genes are MtAPY1;3/MtAPY2, MtAPY1;3/MtAPY1;5 and MtAPY1;2/MtAPY1;3 respectively. In roots, at least four of the MtAPY1 genes are induced transiently within 3 to 6 h by a stress response that seems to be ethylene independent because it occurs after treatment with an ethylene synthesis inhibitor and also in the skl ethylene-insensitive mutant. This response also occurs in roots of the following symbiotic mutants: dmi1, dmi2, dmi3, nsp, hcl, pdl, lin, and skl. No evidence was obtained for a rapid, transient, and specific induction of the MtAPY genes in roots in response to rhizobia or rhizobial lipochitooligosaccharidic Nod factors. Thus, our data suggest that the apyrase like genes, which in several legumes have been implicated to play a role in the legume-rhizobia symbiosis (with some members being described as early nodulin genes), are not regulated symbiotically by rhizobia in M. truncatula. PMID- 12644665 TI - In vivo gas exchange measurement of the site and dynamics of nitrate reduction in soybean. AB - A gas analysis system was built to study the relationship between the reductant cost of NO(3)(-) assimilation and the measured rate of CO(2) and O(2) exchange in roots, leaves, and stems+ petioles of soybean (Glycine max L. Merr. cv Maple glen) plants. The measurements were used to calculate the diverted reductant utilization rate (DRUR = 4*[measured rate of CO(2) + measured rate of O(2)], in moles of high-energy electron [e(-)] per gram per hour) in plants in the presence (N+) and absence (N-) of NO(3)(-). The differences in DRUR between the N+ and N- treatments provided a measure of the NO(3)(-)-coupled DRUR of 25-d-old plants, whereas a (15)NO(3)(-)-enriched nutrient solution was used to obtain an independent measure of the rate of NO(3)(-) assimilation. The measured reductant cost for the whole plant was 9.6 e(-) per N assimilated, a value within the theoretical range of four to 10 e(-) per N assimilated. The results predicted that shoots accounted for about 55% of the whole-plant NO(3)(-) assimilation over the entire day, with shoots dominating in the light, and roots in the dark. The gas analysis approach described here holds promise as a powerful, noninvasive tool to study the regulation of NO(3)(-) assimilation in plant tissue. PMID- 12644666 TI - Secretion of secondary metabolites by ATP-binding cassette transporters in plant cell suspension cultures. PMID- 12644668 TI - The ATP-binding cassette transporters: structure, function, and gene family comparison between rice and Arabidopsis. PMID- 12644669 TI - Analysis of the plastidic phosphate translocator gene family in Arabidopsis and identification of new phosphate translocator-homologous transporters, classified by their putative substrate-binding site. AB - Analysis of the Arabidopsis genome revealed the complete set of plastidic phosphate translocator (pPT) genes. The Arabidopsis genome contains 16 pPT genes: single copies of genes coding for the triose phosphate/phosphate translocator and the xylulose phosphate/phosphate translocator, and two genes coding for each the phosphoenolpyruvate/phosphate translocator and the glucose-6-phosphate/phosphate translocator. A relatively high number of truncated phosphoenolpyruvate/phosphate translocator genes (six) and glucose-6-phosphate/phosphate translocator genes (four) could be detected with almost conserved intron/exon structures as compared with the functional genes. In addition, a variety of PT-homologous (PTh) genes could be identified in Arabidopsis and other organisms. They all belong to the drug/metabolite transporter superfamily showing significant similarities to nucleotide sugar transporters (NSTs). The pPT, PTh, and NST proteins all possess six to eight transmembrane helices. According to the analysis of conserved motifs in these proteins, the PTh proteins can be divided into (a) the lysine (Lys)/arginine group comprising only non-plant proteins, (b) the Lys valine/alanine/glycine group of Arabidopsis proteins, (c) the Lys/asparagine group of Arabidopsis proteins, and (d) the Lys/threonine group of plant and non plant proteins. None of these proteins have been characterized so far. The analysis of the putative substrate-binding sites of the pPT, PTh, and NST proteins led to the suggestion that all these proteins share common substrate binding sites on either side of the membrane each of which contain a conserved Lys residue. PMID- 12644671 TI - Arabidopsis proteins containing similarity to the universal stress protein domain of bacteria. AB - We have collected a set of 44 Arabidopsis proteins with similarity to the USPA (universal stress protein A of Escherichia coli) domain of bacteria. The USPA domain is found either in small proteins, or it makes up the N-terminal portion of a larger protein, usually a protein kinase. Phylogenetic tree analysis based upon a multiple sequence alignment of the USPA domains shows that these domains of protein kinases 1.3.1 and 1.3.2 form distinct groups, as do the protein kinases 1.4.1. This indicates that their USPA domain structures have diverged appreciably and suggests that they may subserve distinct cellular functions. Two USPA fold classes have been proposed: one based on Methanococcus jannaschii MJ0577 (1MJH) that binds ATP, and the other based on the Haemophilus influenzae universal stress protein (1JMV), highly similar to E. coli UspA, which does not bind ATP. A set of common residues involved in ATP binding in 1MJH and conserved in similar bacterial sequences is also found in a distinct cluster of Arabidopsis sequences. Threading analysis, which examines aspects of secondary and tertiary structure, confirms this Arabidopsis sequence cluster as highly similar to 1MJH. This structural approach can distinguish between the characteristic fold differences of 1MJH-like and 1JMV-like bacterial proteins and was used to assign the complete set of candidate Arabidopsis proteins to one of these fold classes. It is clear that all the plant sequences have arisen from a 1MJH-like ancestor. PMID- 12644672 TI - Expression studies of gibberellin oxidases in developing pumpkin seeds. AB - Two cDNA clones, 3-ox and 2-ox, have been isolated from developing pumpkin (Cucurbita maxima) embryos that show significant amino acid homology to gibberellin (GA) 3-oxidases and 2-oxidases, respectively. Recombinant fusion protein of clone 3-ox converted GA(12)-aldehyde, GA(12), GA(15), GA(24), GA(25), and GA(9) to GA(14)-aldehyde, GA(14), GA(37), GA(36), GA(13), and GA(4), respectively. Recombinant 2-ox protein oxidized GA(9), GA(4), and GA(1) to GA(51), GA(34), and GA(8), respectively. Previously cloned GA 7-oxidase revealed additional 3beta-hydroxylation activity of GA(12). Transcripts of this gene were identified in endosperm and embryo of the developing seed by quantitative reverse transcriptase-polymerase chain reaction and localized in protoderm, root apical meristem, and quiescent center by in situ hybridization. mRNA of the previously cloned GA 20-oxidase from pumpkin seeds was localized in endosperm and in tissues of protoderm, ground meristem, and cotyledons of the embryo. However, transcripts of the recently cloned GA 20-oxidase from pumpkin seedlings were found all over the embryo, and in tissues of the inner seed coat at the micropylar end. Previously cloned GA 2beta,3beta-hydroxylase mRNA molecules were specifically identified in endosperm tissue. Finally, mRNA molecules of the 3-ox and 2-ox genes were found in the embryo only. 3-ox transcripts were localized in tissues of cotyledons, protoderm, and inner cell layers of the root apical meristem, and 2-ox transcripts were found in all tissues of the embryo except the root tips. These results indicate tissue-specific GA-biosynthetic pathways operating within the developing seed. PMID- 12644667 TI - Rapid and efficient suppression of gene expression in a single-cell model system, Ceratopteris richardii. PMID- 12644673 TI - The Arabidopsis mutant alh1 illustrates a cross talk between ethylene and auxin. AB - Ethylene or its precursor 1-aminocyclopropane-1-carboxylic acid (ACC) can stimulate hypocotyl elongation in light-grown Arabidopsis seedlings. A mutant, designated ACC-related long hypocotyl 1 (alh1), that displayed a long hypocotyl in the light in the absence of the hormone was characterized. Etiolated alh1 seedlings overproduced ethylene and had an exaggerated apical hook and a thicker hypocotyl, although no difference in hypocotyl length was observed when compared with wild type. Alh1 plants were less sensitive to ethylene, as reflected by reduction of ACC-mediated inhibition of hypocotyl growth in the dark and delay in flowering and leaf senescence. Alh1 also had an altered response to auxin, whereas auxin levels in whole alh1 seedlings remained unaffected. In contrast to wild type, alh1 seedlings showed a limited hypocotyl elongation when treated with indole-3-acetic acid. Alh1 roots had a faster response to gravity. Furthermore, the hypocotyl elongation of alh1 and of ACC-treated wild type was reverted by auxin transport inhibitors. In addition, auxin up-regulated genes were ectopically expressed in hypocotyls upon ACC treatment, suggesting that the ethylene response is mediated by auxins. Together, these data indicate that alh1 is altered in the cross talk between ethylene and auxins, probably at the level of auxin transport. PMID- 12644674 TI - Pto mutants differentially activate Prf-dependent, avrPto-independent resistance and gene-for-gene resistance. AB - Pto confers disease resistance to Pseudomonas syringae pv tomato carrying the cognate avrPto gene. Overexpression of Pto under the cauliflower mosaic virus 35S promoter activates spontaneous lesions and confers disease resistance in tomato (Lycopersicon esculentum) plants in the absence of avrPto. Here, we show that these AvrPto-independent defenses require a functional Prf gene. Several Pto interacting (Pti) proteins are thought to play a role in Pto-mediated defense pathways. To test if interactions with Pti proteins are required for the AvrPto independent defense responses by Pto overexpression, we isolated several Pto mutants that were unable to interact with one or more Pti proteins, but retained normal interaction with AvrPto. Overexpression of two mutants, Pto(G50S) and Pto(R150S), failed to activate AvrPto-independent defense responses or confer enhanced resistance to the virulent P. s. pv tomato. When introduced into plants carrying 35S::Pto, 35S::Pto(G50S) dominantly suppressed the AvrPto-independent resistance caused by former transgene. 35S::Pto(G50S) also blocked the induction of a number of defense genes by the wild-type 35S::Pto. However, 35S::Pto(G50S) and 35S::Pto(R150S) plants were completely resistant to P. s. pv tomato (avrPto), indicating a normal gene-for-gene resistance. Furthermore, 35S::Pto(G50S) plants exhibited normal induction of defense genes in recognition of avrPto. Thus, the AvrPto-independent defense activation and gene-for-gene resistance mediated by Pto are functionally separable. PMID- 12644675 TI - Enhanced selenium tolerance and accumulation in transgenic Arabidopsis expressing a mouse selenocysteine lyase. AB - Selenium (Se) toxicity is thought to be due to nonspecific incorporation of selenocysteine (Se-Cys) into proteins, replacing Cys. In an attempt to direct Se flow away from incorporation into proteins, a mouse (Mus musculus) Se-Cys lyase (SL) was expressed in the cytosol or chloroplasts of Arabidopsis. This enzyme specifically catalyzes the decomposition of Se-Cys into elemental Se and alanine. The resulting SL transgenics were shown to express the mouse enzyme in the expected intracellular location, and to have SL activities up to 2-fold (cytosolic lines) or 6-fold (chloroplastic lines) higher than wild-type plants. Se incorporation into proteins was reduced 2-fold in both types of SL transgenics, indicating that the approach successfully redirected Se flow in the plant. Both the cytosolic and chloroplastic SL plants showed enhanced shoot Se concentrations, up to 1.5-fold compared with wild type. The cytosolic SL plants showed enhanced tolerance to Se, presumably because of their reduced protein Se levels. Surprisingly, the chloroplastic SL transgenics were less tolerant to Se, indicating that (over) production of elemental Se in the chloroplast is toxic. Expression of SL in the cytosol may be a useful approach for the creation of plants with enhanced Se phytoremediation capacity. PMID- 12644670 TI - Analysis of the small GTPase gene superfamily of Arabidopsis. AB - Small GTP-binding proteins regulate diverse processes in eukaryotic cells such as signal transduction, cell proliferation, cytoskeletal organization, and intracellular membrane trafficking. These proteins function as molecular switches that cycle between "active" and "inactive" states, and this cycle is linked to the binding and hydrolysis of GTP. The Arabidopsis genome contains 93 genes that encode small GTP-binding protein homologs. Phylogenetic analysis of these genes shows that plants contain Rab, Rho, Arf, and Ran GTPases, but no Ras GTPases. We have assembled complete lists of these small GTPases families, as well as accessory proteins that control their activity, and review what is known of the functions of individual members of these families in Arabidopsis. We also discuss the possible roles of these GTPases in relation to their similarity to orthologs with known functions and localizations in yeast and/or animal systems. PMID- 12644676 TI - Sterols regulate development and gene expression in Arabidopsis. AB - Sterols are important not only for structural components of eukaryotic cell membranes but also for biosynthetic precursors of steroid hormones. In plants, the diverse functions of sterol-derived brassinosteroids (BRs) in growth and development have been investigated rigorously, yet little is known about the regulatory roles of other phytosterols. Recent analysis of Arabidopsis fackel (fk) mutants and cloning of the FK gene that encodes a sterol C-14 reductase have indicated that sterols play a crucial role in plant cell division, embryogenesis, and development. Nevertheless, the molecular mechanism underlying the regulatory role of sterols in plant development has not been revealed. In this report, we demonstrate that both sterols and BR are active regulators of plant development and gene expression. Similar to BR, both typical (sitosterol and stigmasterol) and atypical (8, 14-diene sterols accumulated in fk mutants) sterols affect the expression of genes involved in cell expansion and cell division. The regulatory function of sterols in plant development is further supported by a phenocopy of the fk mutant using a sterol C-14 reductase inhibitor, fenpropimorph. Although fenpropimorph impairs cell expansion and affects gene expression in a dose dependent manner, neither effect can be corrected by applying exogenous BR. These results provide strong evidence that sterols are essential for normal plant growth and development and that there is likely a BR-independent sterol response pathway in plants. On the basis of the expression of endogenous FK and a reporter gene FK::beta-glucuronidase, we have found that FK is up-regulated by several growth-promoting hormones including brassinolide and auxin, implicating a possible hormone crosstalk between sterol and other hormone-signaling pathways. PMID- 12644677 TI - Transgenic studies on the involvement of cytokinin and gibberellin in male development. AB - Numerous plant hormones interact during plant growth and development. Elucidating the role of these various hormones on particular tissue types or developmental stages has been difficult with exogenous applications or constitutive expression studies. Therefore, we used tissue-specific promoters expressing CKX1 and gai, genes involved in oxidative cytokinin degradation and gibberellin (GA) signal transduction, respectively, to study the roles of cytokinin and GA in male organ development. Accumulation of CKX1 in reproductive tissues of transgenic maize (Zea mays) resulted in male-sterile plants. The male development of these plants was restored by applications of kinetin and thidiazuron. Similarly, expression of gai specifically in anthers and pollen of tobacco (Nicotiana tabacum) and Arabidopsis resulted in the abortion of these respective tissues. The gai-induced male-sterile phenotype exhibited by the transgenic plants was reversible by exogenous applications of kinetin. Our results provide molecular evidence of the involvement of cytokinin and GA in male development and support the hypothesis that the male development is controlled in concert by multiple hormones. These studies also suggest a potential method for generating maintainable male sterility in plants by using existing agrochemicals that would reduce the expense of seed production for existing hybrid crops and provide a method to produce hybrid varieties of traditionally non-hybrid crops. PMID- 12644678 TI - Isolation and properties of floral defensins from ornamental tobacco and petunia. AB - The flowers of the solanaceous plants ornamental tobacco (Nicotiana alata) and petunia (Petunia hybrida) produce high levels of defensins during the early stages of development. In contrast to the well-described seed defensins, these floral defensins are produced as precursors with C-terminal prodomains of 27 to 33 amino acids in addition to a typical secretion signal peptide and central defensin domain of 47 or 49 amino acids. Defensins isolated from N. alata and petunia flowers lack the C-terminal domain, suggesting that it is removed during or after transit through the secretory pathway. Immunogold electron microscopy has been used to demonstrate that the N. alata defensin is deposited in the vacuole. In addition to the eight canonical cysteine residues that define the plant defensin family, the two petunia defensins have an extra pair of cysteines that form a fifth disulfide bond and hence define a new subclass of this family of proteins. Expression of the N. alata defensin NaD1 is predominantly flower specific and is most active during the early stages of flower development. NaD1 transcripts accumulate in the outermost cell layers of petals, sepals, anthers, and styles, consistent with a role in protection of the reproductive organs against potential pathogens. The floral defensins inhibit the growth of Botrytis cinerea and Fusarium oxysporum in vitro, providing further support for a role in protection of floral tissues against pathogen invasion. PMID- 12644679 TI - Genetic diversity of Eurycoma longifolia inferred from single nucleotide polymorphisms. AB - Eurycoma longifolia Jack. is a treelet that grows in the forests of Southeast Asia and is widely used throughout the region because of its reported medicinal properties. Widespread harvesting of wild-grown trees has led to rapid thinning of natural populations, causing a potential decrease in genetic diversity among E. longifolia. Suitable genetic markers would be very useful for propagation and breeding programs to support conservation of this species, although no such markers currently exist. To meet this need, we have applied a genome complexity reduction strategy to identify a series of single nucleotide polymorphisms (SNPs) within the genomes of several E. longifolia accessions. We have found that the occurrence of these SNPs reflects the geographic origins of individual plants and can distinguish different natural populations. This work demonstrates the rapid development of molecular genetic markers in species for which little or no genomic sequence information is available. The SNP markers that we have developed in this study will also be useful for identifying genetic fingerprints that correlate with other properties of E. longifolia, such as high regenerability or the appearance of bioactive metabolites. PMID- 12644680 TI - Plasma membrane H+-ATPase is involved in auxin-mediated cell elongation during wheat embryo development. AB - Previous investigations suggested that specific auxin spatial distribution due to auxin movements to particular embryonic regions was important for normal embryonic pattern formation. To gain information on the molecular mechanism(s) by which auxin acts to direct pattern formation in specific embryonic regions, the role of a plasma membrane (PM) ATPase was evaluated as downstream target of auxin in the present study. Western-blot analysis revealed that the PM H(+)-ATPase expression level was significantly increased by auxin in wheat (Triticum aestivum) embryos (two-three times increase). In bilaterally symmetrical embryos, the spatial expression pattern of the PM H(+)-ATPase correlates with the distribution pattern of the auxin analog, tritiated 5-azidoindole-3-acetic acid. A strong immunosignal was observed in the abaxial epidermis of the scutellum and in the epidermal cells at the distal tip of this organ. Pseudoratiometric analysis using a fluorescent pH indicator showed that the pH in the apoplast of the cells expressing the PM H(+)-ATPase was in average more acidic than the apoplastic pH of nonexpressing cells. Cellulose staining of living embryos revealed that cells of the scutellum abaxial epidermis expressing the ATPase were longer than the scutellum adaxial epidermal cells, where the protein was not expressed. Our data indicate that auxin activates the proton pump resulting in apoplastic acidification, a process contributing to cell wall loosening and elongation of the scutellum. Therefore, we suggest that the PM H(+)-ATPase is a component of the auxin-signaling cascade that may direct pattern formation in embryos. PMID- 12644681 TI - Whole-genome comparison of leucine-rich repeat extensins in Arabidopsis and rice. A conserved family of cell wall proteins form a vegetative and a reproductive clade. AB - We have searched the Arabidopsis and rice (Oryza sativa) genomes for homologs of LRX1, an Arabidopsis gene encoding a novel type of cell wall protein containing a leucine-rich repeat (LRR) and an extensin domain. Eleven and eight LRX (LRR/EXTENSIN) genes have been identified in these two plant species, respectively. The LRX gene family encodes proteins characterized by a short N terminal domain, a domain with 10 LRRs, a cysteine-rich motif, and a variable C terminal extensin-like domain. Phylogenetic analysis performed on the conserved domains indicates the existence of two major clades of LRX proteins that arose before the eudicot/monocot divergence and then diversified independently in each lineage. In Arabidopsis, gene expression studies by northern hybridization and promoter::uidA fusions showed that the two phylogenetic clades represent a specialization into "reproductive" and "vegetative" LRXs. The four Arabidopsis genes of the "reproductive" clade are specifically expressed in pollen, whereas the seven "vegetative" genes are predominantly expressed in various sporophytic tissues. This separation into two expression classes is also supported by previous studies on maize (Zea mays) and tomato (Lycopersicon esculentum) LRX homologs and by information on available rice ESTs. The strong conservation of the amino acids responsible for the putative recognition specificity of the LRR domain throughout the family suggests that the LRX proteins interact with similar ligands. PMID- 12644682 TI - Auxin signaling in Arabidopsis leaf vascular development. AB - A number of observations have implicated auxin in the formation of vascular tissues in plant organs. These include vascular strand formation in response to local auxin application, the effects of impaired auxin transport on vascular patterns and suggestive phenotypes of Arabidopsis auxin response mutants. In this study, we have used molecular markers to visualize auxin response patterns in developing Arabidopsis leaves as well as Arabidopsis mutants and transgenic plants to trace pathways of auxin signal transduction controlling the expression of early procambial genes. We show that in young Arabidopsis leaf primordia, molecular auxin response patterns presage sites of procambial differentiation. This is the case not only in normal development but also upon experimental manipulation of auxin transport suggesting that local auxin signals are instrumental in patterning Arabidopsis leaf vasculature. We further found that the activity of the Arabidopsis gene MONOPTEROS, which is required for proper vascular differentiation, is also essential in a spectrum of auxin responses, which include the regulation of rapidly auxin-inducible AUX/IAA genes, and discovered the tissue-specific vascular expression profile of the class I homeodomain-leucine zipper gene, AtHB20. Interestingly, MONOPTEROS activity is a limiting factor in the expression of AtHB8 and AtHB20, two genes encoding transcriptional regulators expressed early in procambial development. Our observations connect general auxin signaling with early controls of vascular differentiation and suggest molecular mechanisms for auxin signaling in patterned cell differentiation. PMID- 12644683 TI - Phytochromes B, D, and E act redundantly to control multiple physiological responses in Arabidopsis. AB - Phytochrome-mediated perception of the ratio of red to far-red wavelengths in the ambient light environment is fundamental to plant growth and development. Such monitoring enables plants to detect neighboring vegetation and initiate avoidance responses, thus conferring considerable selective advantage. The shade avoidance syndrome in plants is characterized by elongation growth and early flowering, responses that are fully induced by end-of-day far-red light treatments. Elucidating the roles of individual phytochromes in mediating responses to red to far-red has however always been confounded by synergistic and mutually antagonistic coactions between family members. The creation of triple and quadruple mutants in Arabidopsis, deficient in multiple phytochromes, has revealed functional redundancy between phyB, D, and E in controlling flowering time, leaf development, and regulation of the homeobox gene, ATHB-2. In addition, mutant analysis suggests a possible novel role for phyC in suppressing ATHB-2 transcription in the light. PMID- 12644684 TI - Expression of a GALACTINOL SYNTHASE gene in tomato seeds is up-regulated before maturation desiccation and again after imbibition whenever radicle protrusion is prevented. AB - Raffinose family oligosaccharides (RFOs) have been implicated in mitigating the effects of environmental stresses on plants. In seeds, proposed roles for RFOs include protecting cellular integrity during desiccation and/or imbibition, extending longevity in the dehydrated state, and providing substrates for energy generation during germination. A gene encoding galactinol synthase (GOLS), the first committed enzyme in the biosynthesis of RFOs, was cloned from tomato (Lycopersicon esculentum Mill. cv Moneymaker) seeds, and its expression was characterized in tomato seeds and seedlings. GOLS (LeGOLS-1) mRNA accumulated in developing tomato seeds concomitant with maximum dry weight deposition and the acquisition of desiccation tolerance. LeGOLS-1 mRNA was present in mature, desiccated seeds but declined within 8 h of imbibition in wild-type seeds. However, LeGOLS-1 mRNA accumulated again in imbibed seeds prevented from completing germination by dormancy or water deficit. Gibberellin-deficient (gib 1) seeds maintained LeGOLS-1 mRNA amounts after imbibition unless supplied with gibberellin, whereas abscisic acid (ABA) did not prevent the loss of LeGOLS-1 mRNA from wild-type seeds. The presence of LeGOLS-1 mRNA in ABA-deficient (sitiens) tomato seeds indicated that wild-type amounts of ABA are not necessary for its accumulation during seed development. In all cases, LeGOLS-1 mRNA was most prevalent in the radicle tip. LeGOLS-1 mRNA accumulation was induced by dehydration but not by cold in germinating seeds, whereas both stresses induced LeGOLS-1 mRNA accumulation in seedling leaves. The physiological implications of LeGOLS-1 expression patterns in seeds and leaves are discussed in light of the hypothesized role of RFOs in plant stress tolerance. PMID- 12644685 TI - Enhanced gravitropism of roots with a disrupted cap actin cytoskeleton. AB - The actin cytoskeleton has been proposed to be a major player in plant gravitropism. However, understanding the role of actin in this process is far from complete. To address this problem, we conducted an analysis of the effect of Latrunculin B (Lat B), a potent actin-disrupting drug, on root gravitropism using various parameters that included detailed curvature kinetics, estimation of gravitropic sensitivity, and monitoring of curvature development after extended clinorotation. Lat B treatment resulted in a promotion of root curvature after a 90 degrees reorientation in three plant species tested. More significantly, the sensitivity of maize (Zea mays) roots to gravity was enhanced after actin disruption, as determined from a comparison of presentation time of Lat B-treated versus untreated roots. A short 10-min gravistimulus followed by extended rotation on a 1-rpm clinostat resulted in extensive gravitropic responses, manifested as curvature that often exceeded 90 degrees. Application of Lat B to the cap or elongation zone of maize roots resulted in the disruption of the actin cytoskeleton, which was confined to the area of localized Lat B application. Only roots with Lat B applied to the cap displayed the strong curvature responses after extended clinorotation. Our study demonstrates that disrupting the actin cytoskeleton in the cap leads to the persistence of a signal established by a previous gravistimulus. Therefore, actin could function in root gravitropism by providing a mechanism to regulate the proliferation of a gravitropic signal originating from the cap to allow the root to attain its correct orientation or set point angle. PMID- 12644686 TI - O-glucosylation of cis-zeatin in maize. Characterization of genes, enzymes, and endogenous cytokinins. AB - trans-Zeatin is a major and ubiquitous cytokinin in higher plants. cis-Zeatin has traditionally been viewed as an adjunct with low activity and rare occurrence. Recent reports of cis-zeatin and its derivatives as the predominant cytokinin components in some plant tissues may call for a different perspective on cis isomers. The existence of a maize (Zea mays) gene (cisZOG1) encoding an O glucosyltransferase specific to cis-zeatin (R.C. Martin, M.C. Mok, J.E. Habben, D.W.S. Mok [2001] Proc Natl Acad Sci USA 98: 5922-5926) lends further support to this view. Results described here include the isolation of a second maize cisZOG gene, differential expression of cisZOG1 and cisZOG2, and identification of substantial amounts of cis-isomers in maize tissues. The open reading frame of cisZOG2 has 98.3% identity to cisZOG1 at the nucleotide level and 97.8% at the amino acid level. The upstream regions contain common and unique segments. The recombinant enzymes have similar properties, K(m) values of 46 and 96 microM, respectively, for cis-zeatin and a pH optimum of 7.5. Other cytokinins, including N(6)-(delta(2)-isopentenyl)adenine, trans-zeatin, benzyladenine, kinetin, and thidiazuron inhibited the reaction. Expression of cisZOG1 was high in maize roots and kernels, whereas cisZOG2 expression was high in roots but low in kernels. cis Zeatin, cis-zeatin riboside, and their O-glucosides were detected in all maize tissues, with immature kernels containing very high levels of the O-glucoside of cis-zeatin riboside. The results are a clear indication that O-glucosylation of cis-zeatin is a natural metabolic process in maize. Whether cis-zeatin serves as a precursor to the active trans-isomer or has any other unique function remains to be demonstrated. PMID- 12644688 TI - A novel Cl- inward-rectifying current in the plasma membrane of the calcifying marine phytoplankton Coccolithus pelagicus. AB - We investigated the membrane properties and dominant ionic conductances in the plasma membrane of the calcifying marine phytoplankton Coccolithus pelagicus using the patch-clamp technique. Whole-cell recordings obtained from decalcified cells revealed a dominant anion conductance in response to membrane hyperpolarization. Ion substitution showed that the anion channels were selective for Cl(-) and Br(-) over other anions, and the sensitivity to the stilbene derivative 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid, ethacrynic acid, and Zn(2+) revealed a pharmacological profile typical of many plant and animal anion channels. Voltage activation and kinetic characteristics of the C. pelagicus Cl(-) channel are consistent with a novel function in plants as the inward rectifier that tightly regulates membrane potential. Membrane depolarization gave rise to nonselective cation currents and in some cases evoked action potential currents. We propose that these major ion conductances play an essential role in membrane voltage regulation that relates to the unique transport physiology of these calcifying phytoplankton. PMID- 12644687 TI - Regulation of root elongation under phosphorus stress involves changes in ethylene responsiveness. AB - We characterized the growth of the primary root of Arabidopsis under phosphorus sufficiency (1 mM phosphate) and deficiency (1 microM phosphate), focusing on the role of ethylene. We quantified the spatial profile of relative elongation with a novel method based on image processing, as well as the production rates of cortical cells, trichoblasts, and atrichoblasts. Phosphorus deficiency moderately decreased the maximal rate of relative elongation, shortened the growth zone, and decreased the production rate of both epidermal cell types but not of cortical cells. Inhibiting ethylene production (with aminoethoxyvinyl-glycine) or action (with 1-methylcyclopropene) increased elongation in high phosphorus and decreased it in low phosphorus. That these effects were specific to ethylene was confirmed by negating the effect of inhibited ethylene production with simultaneous treatment with an ethylene precursor (1-aminocyclopropane-1-carboxylic acid). Under both phosphorus regimes, ethylene regulated the maximal rate of relative elongation rather than the size of the growth zone. In addition, inhibiting ethylene action in high versus low phosphorus elicited opposite responses for the position of root hair initiation and for the production rates of cortex cells and atrichoblasts. We conclude that the root system acclimates to phosphorus deficiency by changing the signal transduction pathway connecting ethylene levels to growth and division. PMID- 12644689 TI - 4-Coumarate:coenzyme A ligase has the catalytic capacity to synthesize and reuse various (di)adenosine polyphosphates. AB - 4-Coumarate:coenzyme A ligase (4CL) is known to activate cinnamic acid derivatives to their corresponding coenzyme A esters. As a new type of 4CL catalyzed reaction, we observed the synthesis of various mono- and diadenosine polyphosphates. Both the native 4CL2 isoform from Arabidopsis (At4CL2 wild type) and the At4CL2 gain of function mutant M293P/K320L, which exhibits the capacity to use a broader range of phenolic substrates, catalyzed the synthesis of adenosine 5'-tetraphosphate (p(4)A) and adenosine 5'-pentaphosphate when incubated with MgATP(-2) and tripolyphosphate or tetrapolyphosphate (P(4)), respectively. Diadenosine 5',5''',-P(1),P(4)-tetraphosphate represented the main product when the enzymes were supplied with only MgATP(2-). The At4CL2 mutant M293P/K320L was studied in more detail and was also found to catalyze the synthesis of additional dinucleoside polyphosphates such as diadenosine 5',5''' P(1),P(5)-pentaphosphate and dAp(4)dA from the appropriate substrates, p(4)A and dATP, respectively. Formation of Ap(3)A from ATP and ADP was not observed with either At4CL2 variant. In all cases analyzed, (di)adenosine polyphosphate synthesis was either strictly dependent on or strongly stimulated by the presence of a cognate cinnamic acid derivative. The At4CL2 mutant enzyme K540L carrying a point mutation in the catalytic center that is critical for adenylate intermediate formation was inactive in both p(4)A and diadenosine 5',5''', P(1),P(4)-tetraphosphate synthesis. These results indicate that the cinnamoyl adenylate intermediate synthesized by At4CL2 not only functions as an intermediate in coenzyme A ester formation but can also act as a cocatalytic AMP donor in (di)adenosine polyphosphate synthesis. PMID- 12644690 TI - Phytochromes A and B mediate red-light-induced positive phototropism in roots. AB - The interaction of tropisms is important in determining the final growth form of the plant body. In roots, gravitropism is the predominant tropistic response, but phototropism also plays a role in the oriented growth of roots in flowering plants. In blue or white light, roots exhibit negative phototropism that is mediated by the phototropin family of photoreceptors. In contrast, red light induces a positive phototropism in Arabidopsis roots. Because this red-light induced response is weak relative to both gravitropism and negative phototropism, we used a novel device to study phototropism without the complications of a counteracting gravitational stimulus. This device is based on a computer controlled system using real-time image analysis of root growth and a feedback regulated rotatable stage. Our data show that this system is useful to study root phototropism in response to red light, because in wild-type roots, the maximal curvature detected with this apparatus is 30 degrees to 40 degrees, compared with 5 degrees to 10 degrees without the feedback system. In positive root phototropism, sensing of red light occurs in the root itself and is not dependent on shoot-derived signals resulting from light perception. Phytochrome (Phy)A and phyB were severely impaired in red-light-induced phototropism, whereas the phyD and phyE mutants were normal in this response. Thus, PHYA and PHYB play a key role in mediating red-light-dependent positive phototropism in roots. Although phytochrome has been shown to mediate phototropism in some lower plant groups, this is one of the few reports indicating a phytochrome-dependent phototropism in flowering plants. PMID- 12644691 TI - Regulation of the alternative oxidase Aox1 gene in Chlamydomonas reinhardtii. Role of the nitrogen source on the expression of a reporter gene under the control of the Aox1 promoter. AB - In higher plants, various developmental and environmental conditions enhance expression of the alternative oxidase (AOX), whereas its induction in fungi is mainly dependent on cytochrome pathway restriction and triggering by reactive oxygen species. The AOX of the unicellular green alga Chlamydomonas reinhardtii is encoded by two different genes, the Aox1 gene being much more transcribed than Aox2. To analyze the transcriptional regulation of Aox1, we have fused its 1.4-kb promoter region to the promoterless arylsulfatase (Ars) reporter gene and measured ARS enzyme activities in transformants carrying the chimeric construct. We show that the Aox1 promoter is generally unresponsive to a number of known AOX inducers, including stress agents, respiratory inhibitors, and metabolites, possibly because the AOX activity is constitutively high in the alga. In contrast, the Aox1 expression is strongly dependent on the nitrogen source, being down-regulated by ammonium and stimulated by nitrate. Inactivation of nitrate reductase leads to a further increase of expression. The stimulation by nitrate also occurs at the AOX protein and respiratory levels. A deletion analysis of the Aox1 promoter region demonstrates that a short upstream segment (-253 to +59 with respect to the transcription start site) is sufficient to ensure gene expression and regulation, but that distal elements are required for full gene expression. The observed pattern of AOX regulation points to the possible interaction between chloroplast and mitochondria in relation to a potential increase of photogenerated ATP when nitrate is used as a nitrogen source. PMID- 12644692 TI - One-carbon metabolism in plants. Regulation of tetrahydrofolate synthesis during germination and seedling development. AB - Tetrahydrofolate (THF) is a central cofactor for one-carbon transfer reactions in all living organisms. In this study, we analyzed the expression of dihydropterin pyrophosphokinase-dihydropteroate synthase (HPPK-DHPS) in pea (Pisum sativum) organs during development, and so the capacity to synthesize dihydropteroate, an intermediate in the de novo THF biosynthetic pathway. During seedling development, all of the examined organs/tissues contain THF coenzymes, collectively termed folate, and express the HPPK-DHPS enzyme. This suggests that each organ/tissue is autonomous for the synthesis of THF. During germination, folate accumulates in cotyledons and embryos, but high amounts of HPPK-DHPS are only observed in embryos. During organ differentiation, folate is synthesized preferentially in highly dividing tissues and in photosynthetic leaves. This is associated with high levels of the HPPK-DHPS mRNA and protein, and a pool of folate 3- to 5-fold higher than in the rest of the plant. In germinating embryos and in meristematic tissues, the high capacity to synthesize and accumulate folate correlates with the general resumption of cell metabolism and the high requirement for nucleotide synthesis, major cellular processes involving folate coenzymes. The particular status of folate synthesis in leaves is related to light. Thus, when illuminated, etiolated leaves gradually accumulate the HPPK DHPS enzyme and folate. This suggests that folate synthesis plays an important role in the transition from heterotrophic to photoautotrophic growth. Analysis of the intracellular distribution of folate in green and etiolated leaves indicates that the coenzymes accumulate mainly in the cytosol, where they can supply the high demand for methyl groups. PMID- 12644693 TI - Biphasic superoxide generation in potato tubers. A self-amplifying response to stress. AB - Potato (Solanum tuberosum) cultivars differ quantitatively in their responses to mechanical stress including the ability to synthesize melanin pigments in tuber tissues. Investigations into the cellular events induced by mechanical stress on tuber tissues have shown that an early cellular response is a significant and rapid synthesis of superoxide radicals. This burst of radical production distinctively displays a reproducible biphasic pattern over time with peaks of generation at 2 and 5 h. A concomitant consequence of the generation of these free radicals is elevated levels of oxidatively modified tuber proteins. Both radical generation and protein modification vary between cultivars but both are directly proportional to the amount of melanin pigments produced. Cell-free extracts of mechanically stressed tissues, pectic fragments, and scission products generated from cell walls are able to induce superoxide generation in non-stressed tissues, indicating the participation of a biologically active factor that induces a further a phase of radical synthesis. PMID- 12644694 TI - CsAGP1, a gibberellin-responsive gene from cucumber hypocotyls, encodes a classical arabinogalactan protein and is involved in stem elongation. AB - Fluorescence differential display was used to isolate the gibberellin (GA) responsive gene, CsAGP1, from cucumber (Cucumis sativus) hypocotyls. A sequence analysis of CsAGP1 indicated that the gene putatively encodes a "classical" arabinogalactan protein (AGP) in cucumber. Transgenic tobacco (Nicotiana tabacum) plants overexpressing CsAGP1 under the control of the cauliflower mosaic virus 35S promoter produced a Y(betaGlc)(3)-reactive proteoglycan in addition to AGPs present in wild-type tobacco plants. Immuno-dot blotting of the product, using anti-AGP antibodies, showed that the CsAGP1 protein had the AGP epitopes common to AGP families. The transcription level of CsAGP1 in cucumber hypocotyls increased in response not only to GA but also to indole-3-acetic acid. Although CsAGP1 is expressed in most vegetative tissues of cucumber, including the shoot apices and roots, the GA treatment resulted in an increase in the mRNA level of CsAGP1 only in the upper part of the hypocotyls. Y(betaGlc)(3), which selectively binds AGPs, inhibited the hormone-promoted elongation of cucumber seedling hypocotyls. Transgenic plants ectopically expressing CsAGP1 showed a taller stature and earlier flowering than the wild-type plants. These observations suggest that CsAGP1 is involved in stem elongation. PMID- 12644695 TI - Chemical form and distribution of selenium and sulfur in the selenium hyperaccumulator Astragalus bisulcatus. AB - In its natural habitat, Astragalus bisulcatus can accumulate up to 0.65% (w/w) selenium (Se) in its shoot dry weight. X-ray absorption spectroscopy has been used to examine the selenium biochemistry of A. bisulcatus. High concentrations of the nonprotein amino acid Se-methylseleno-cysteine (Cys) are present in young leaves of A. bisulcatus, but in more mature leaves, the Se-methylseleno-Cys concentration is lower, and selenate predominates. Seleno-Cys methyltransferase is the enzyme responsible for the biosynthesis of Se-methylseleno-Cys from seleno Cys and S-methyl-methionine. Seleno-Cys methyltransferase is found to be expressed in A. bisulcatus leaves of all ages, and thus the biosynthesis of Se methylseleno-Cys in older leaves is limited earlier in the metabolic pathway, probably by an inability to chemically reduce selenate. A comparative study of sulfur (S) and Se in A. bisulcatus using x-ray absorption spectroscopy indicates similar trends for oxidized and reduced Se and S species, but also indicates that the proportions of these differ significantly. These results also indicate that sulfate and selenate reduction are developmentally correlated, and they suggest important differences between S and Se biochemistries. PMID- 12644697 TI - The role of biotin in regulating 3-methylcrotonyl-coenzyme a carboxylase expression in Arabidopsis. AB - As a catalytic cofactor, biotin has a critical role in the enzymological mechanism of a number of enzymes that are essential in both catabolic and anabolic metabolic processes. In this study we demonstrate that biotin has additional non-catalytic functions in regulating gene expression in plants, which are biotin autotrophic organisms. Biotin controls expression of the biotin containing enzyme, methylcrotonyl-coenzyme A (CoA) carboxylase by modulating the transcriptional, translational and/or posttranslational regulation of the expression of this enzyme. The bio1 mutant of Arabidopsis, which is blocked in the de novo biosynthesis of biotin, was used to experimentally alter the biotin status of this organism. In response to the bio1-associated depletion of biotin, the normally biotinylated A-subunit of methylcrotonyl-CoA carboxylase (MCCase) accumulates in its inactive apo-form, and both MCCase subunits hyperaccumulate. This hyperaccumulation occurs because the translation of each subunit mRNA is enhanced and/or because the each protein subunit becomes more stable. In addition, biotin affects the accumulation of distinct charge isoforms of MCCase. In contrast, in response to metabolic signals arising from the alteration in the carbon status of the organism, biotin modulates the transcription of the MCCase genes. These experiments reveal that in addition to its catalytic role as an enzyme cofactor, biotin has multiple roles in regulating gene expression. PMID- 12644696 TI - Root factors induce mitochondrial-related gene expression and fungal respiration during the developmental switch from asymbiosis to presymbiosis in the arbuscular mycorrhizal fungus Gigaspora rosea. AB - During spore germination, arbuscular mycorrhizal (AM) fungi show limited hyphal development in the absence of a host plant (asymbiotic). In the presence of root exudates, they switch to a new developmental stage (presymbiotic) characterized by extensive hyphal branching. Presymbiotic branching of the AM fungus Gigaspora rosea was induced in liquid medium by a semipurified exudate fraction from carrot (Daucus carota) root organ cultures. Changes in RNA accumulation patterns were monitored by differential display analysis. Differentially appearing cDNA fragments were cloned and further analyzed. Five cDNA fragments could be identified that show induced RNA accumulation 1 h after the addition of root exudate. Sequence similarities of two fragments to mammalian Nco4 and mitochondrial rRNA genes suggested that root exudates could influence fungal respiratory activity. To support this hypothesis, additional putative mitochondrial related-genes were shown to be induced by root exudates. These genes were identified after subtractive hybridization and putatively encode a pyruvate carboxylase and a mitochondrial ADP/ATP translocase. The gene GrosPyc1 for the pyruvate carboxylase was studied in more detail by cloning a cDNA and by quantifying its RNA accumulation. The hypothesis that respiratory activity of AM fungi is stimulated by root exudates was confirmed by physiological and cytological analyses in G. rosea and Glomus intraradices. Oxygen consumption and reducing activity of both fungi was induced after 3 and 2 h of exposition with the root factor, respectively, and the first respiration activation was detected in G. intraradices after approximately 90 min. In addition, changes in mitochondrial morphology, orientation, and overall biomass were detected in G. rosea after 4 h. In summary, the root-exuded factor rapidly induces the expression of certain fungal genes and, in turn, fungal respiratory activity before intense branching. This defines the developmental switch from asymbiosis to presymbiosis, first by gene activation (0.5-1 h), subsequently on the physiological level (1.5-3 h), and finally as a morphological response (after 5 h). PMID- 12644698 TI - Tobacco transgenic lines that express fenugreek galactomannan galactosyltransferase constitutively have structurally altered galactomannans in their seed endosperm cell walls. AB - Galactomannans [(1-->6)-alpha-D-galactose (Gal)-substituted (1-->4)-beta-D mannans] are major cell wall storage polysaccharides in the endosperms of some seeds, notably the legumes. Their biosynthesis in developing legume seeds involves the functional interaction of two membrane-bound glycosyltransferases, mannan synthase (MS) and galactomannan galactosyltransferase (GMGT). MS catalyzes the elongation of the mannan backbone, whereas GMGT action determines the distribution and amount of Gal substitution. Fenugreek (Trigonella foenum graecum) forms a galactomannan with a very high degree of Gal substitution (Man/Gal = 1.1), and its GMGT has been characterized. We now report that the endosperm cell walls of the tobacco (Nicotiana tabacum) seed are rich in a galactomannan with a very low degree of Gal substitution (Man/Gal about 20) and that its depositional time course is closely correlated with membrane-bound MS and GMGT activities. Furthermore, we demonstrate that seeds from transgenic tobacco lines that express fenugreek GMGT constitutively in membrane-bound form have endosperm galactomannans with increased average degrees of Gal substitution (Man/Gal about 10 in T(1) generation seeds and about 7.5 in T(2) generation seeds). Membrane-bound enzyme systems from transgenic seed endosperms form galactomannans in vitro that are more highly Gal substituted than those formed by controls under identical conditions. To our knowledge, this is the first report of structural manipulation of a plant cell wall polysaccharide in transgenic plants via a biosynthetic membrane-bound glycosyltransferase. PMID- 12644699 TI - Carbon export from arbuscular mycorrhizal roots involves the translocation of carbohydrate as well as lipid. AB - Arbuscular mycorrhizal (AM) fungi take up photosynthetically fixed carbon from plant roots and translocate it to their external mycelium. Previous experiments have shown that fungal lipid synthesized from carbohydrate in the root is one form of exported carbon. In this study, an analysis of the labeling in storage and structural carbohydrates after (13)C(1) glucose was provided to AM roots shows that this is not the only pathway for the flow of carbon from the intraradical to the extraradical mycelium (ERM). Labeling patterns in glycogen, chitin, and trehalose during the development of the symbiosis are consistent with a significant flux of exported glycogen. The identification, among expressed genes, of putative sequences for glycogen synthase, glycogen branching enzyme, chitin synthase, and for the first enzyme in chitin synthesis (glutamine fructose 6-phosphate aminotransferase) is reported. The results of quantifying glycogen synthase gene expression within mycorrhizal roots, germinating spores, and ERM are consistent with labeling observations using (13)C-labeled acetate and glycerol, both of which indicate that glycogen is synthesized by the fungus in germinating spores and during symbiosis. Implications of the labeling analyses and gene sequences for the regulation of carbohydrate metabolism are discussed, and a 4-fold role for glycogen in the AM symbiosis is proposed: sequestration of hexose taken from the host, long-term storage in spores, translocation from intraradical mycelium to ERM, and buffering of intracellular hexose levels throughout the life cycle. PMID- 12644700 TI - Control of steroid, heme, and carcinogen metabolism by nuclear pregnane X receptor and constitutive androstane receptor. AB - Through a multiplex promoter spanning 218 kb, the phase II UDP glucuronosyltransferase 1A (UGT1) gene encodes at least eight differently regulated mRNAs whose protein products function as the principal means to eliminate a vast array of steroids, heme metabolites, environmental toxins, and drugs. The orphan nuclear receptors pregnane X receptor (PXR) and constitutive androstane receptor (CAR) were originally identified as sensors able to respond to numerous environmentally derived foreign compounds (xenobiotics) to promote detoxification by phase I cytochrome P450 genes. In this report, we show that both receptors can induce specific UGT1A isoforms including those involved in estrogen, thyroxin, bilirubin, and carcinogen metabolism. Transgenic mice expressing a constitutively active form of human PXR show markedly increased UGT activity toward steroid, heme, and carcinogens, enhanced bilirubin clearance, as well as massively increased steroid clearance. The ability of PXR and constitutive androstane receptor and their ligands to transduce both the phase I and phase II adaptive hepatic response defines a unique transcriptional interface that bridges the ingestion and metabolism of environmental compounds to body physiology. PMID- 12644701 TI - Lanthanide-binding helix-turn-helix peptides: solution structure of a designed metallonuclease. AB - A designed lanthanide-binding chimeric peptide based on the strikingly similar geometries of the EF-hand and helix-turn-helix (HTH) motifs was investigated by NMR and CD spectroscopy and found to retain the same overall solution structure of the parental motifs. CD spectroscopy showed that the 33-mer peptide P3W folds on binding lanthanides, with an increase in alpha-helicity from 20% in the absence of metal to 38% and 35% in the presence of excess Eu(III) and La(III) ions, respectively. The conditional binding affinities of P3W for La(III) (5.9 +/ 0.3 microM) and for Eu(III) (6.2 +/- 0.3 microM) (pH 7.8, 5 mM Tris) were determined by tryptophan fluorescence titration. The La(III) complex of peptide P3, which differs from P3W by only one Trp-to-His substitution, has much less signal dispersion in the proton NMR spectra than LaP3W, indicating that the Trp residue is a critical hydrophobic anchor for maintaining a well-folded helix-turn helix structure. A chemical-shift index analysis indicates the metallopeptide has a helix-loop-helix secondary structure. A structure calculated by using nuclear Overhauser effect and other NMR constraints reveals that P3W not only has a tightly folded metal-binding loop but also retains the alpha-alpha corner supersecondary structure of the parental motifs. Although the solution structure is undefined at both the N and C termini, the NMR structure confirms the successful incorporation of a metal-binding loop into a HTH sequence. PMID- 12644702 TI - Rapid evolution of the neutralizing antibody response to HIV type 1 infection. AB - A recombinant virus assay was used to characterize in detail neutralizing antibody responses directed at circulating autologous HIV in plasma. Examining serial plasma specimens in a matrix format, most patients with primary HIV infection rapidly generated significant neutralizing antibody responses to early (0-39 months) autologous viruses, whereas responses to laboratory and heterologous primary strains were often lower and delayed. Plasma virus continually and rapidly evolved to escape neutralization, indicating that neutralizing antibody exerts a level of selective pressure that has been underappreciated based on earlier, less comprehensive characterizations. These data argue that neutralizing antibody responses account for the extensive variation in the envelope gene that is observed in the early months after primary HIV infection. PMID- 12644703 TI - Induction of G1 cycle arrest in T lymphocytes results in increased extracellular levels of beta-chemokines: a strategy to inhibit R5 HIV-1. AB - The beta-chemokines RANTES (regulated on activation, normal T cell expressed and secreted), macrophage inflammatory protein-1alpha (MIP-1alpha), and MIP-1beta are the natural ligands of the HIV-1 coreceptor CCR5 and compete with the virus for receptor binding. We show that secretion of the beta-chemokines by activated lymphocytes starts before cellular DNA synthesis is detected and demonstrate that transient prolongation of the G(1) phase of the cell cycle by treatment with cytostatic drugs results in increased levels of the three chemokines in culture supernatants. Supernatants collected from peripheral blood mononuclear cells exposed to hydroxyurea, which arrests the cell cycle in late G(1), contained high levels of beta-chemokines. These supernatants were able to inhibit HIV-1 replication when added to cultures of infected lymphocytes. The observed antiviral effect likely was due to the increased levels of beta-chemokines RANTES, MIP-1alpha, and MIP-1beta because (i) supernatants greatly inhibited the replication of HIV-1 BaL, whereas they affected HIV-1 IIIb replication only slightly; (ii) neutralizing antibodies against the chemokines abrogated the antiviral effect of the supernatants; and (iii) the hydroxyurea concentrations shown to up-regulate chemokine levels were not sufficient to inhibit virus replication by depletion of intracellular nucleotide pools. Although antiviral properties have been reported previously for the cytostatic agents shown here to up-regulate beta-chemokine levels, our results provide an additional mechanism by which these drugs may exert antiviral activity. In summary, increased extracellular levels of anti-HIV-1 beta-chemokines resulting from transient prolongation of the G(1) phase of the lymphocyte cell cycle by treatment with cytostatic drugs may help to control the replication of CCR5-using strains of HIV 1. PMID- 12644704 TI - Induction of bilirubin clearance by the constitutive androstane receptor (CAR). AB - Bilirubin clearance is one of the numerous important functions of the liver. Defects in this process result in jaundice, which is particularly common in neonates. Elevated bilirubin levels can be decreased by treatment with phenobarbital. Because the nuclear hormone receptor constitutive androstane receptor (CAR) mediates hepatic effects of this xenobiotic inducer, we hypothesized that CAR could be a regulator of bilirubin clearance. Activation of the nuclear hormone receptor CAR increases hepatic expression of each of five components of the bilirubin-clearance pathway. This induction is absent in homozygous CAR null mice but is observed in mice expressing human CAR instead of mouse CAR. Pretreatment with xenobiotic inducers markedly increases the rate of clearance of an exogenous bilirubin load in wild-type but not CAR knockout animals. Bilirubin itself can also activate CAR, and mice lacking CAR are defective in clearing chronically elevated bilirubin levels. Unexpectedly, CAR expression is very low in livers of neonatal mice and humans. We conclude that CAR directs a protective response to elevated bilirubin levels and suggest that a functional deficit of CAR activity may contribute to neonatal jaundice. PMID- 12644705 TI - Efficient active transport of gene nanocarriers to the cell nucleus. AB - The intracellular transport of therapeutic gene carriers is poorly understood, limiting the rational design of efficient new vectors. We used live-cell real time multiple particle tracking to quantify the intracellular transport of hundreds of individual nonviral DNA nanocarriers with 5-nm and 33-ms resolution. Unexpected parallels between several of nature's most efficient DNA viruses and nonviral polyethylenimine/DNA nanocomplexes were revealed to include motor protein-driven transport through the cytoplasm toward the nucleus on microtubules. Active gene carrier transport led to efficient perinuclear accumulation within minutes. The results provide direct evidence to dispute the common belief that the efficiency of nonviral gene carriers is dramatically reduced because of the need for their relatively slow random diffusion through the cell cytoplasm to the nucleus and, instead, focuses the attention of rational carrier design on overcoming barriers downstream of perinuclear accumulation. PMID- 12644706 TI - Introduction and characterization of a functionally linked metal ion binding site at the exposed heme edge of myoglobin. AB - A binding site for metal ions has been created on the surface of horse heart myoglobin (Mb) near the heme 6-propionate group by replacing K45 and K63 with glutamyl residues. One-dimensional (1)H NMR spectroscopy indicates that Mn(2+) binds in the vicinity of the heme 6-propionate as anticipated, and potentiometric titrations establish that the affinity of the new site for Mn(2+) is 1.28(4) x 10(4) M(-1) (pH 6.96, ionic strength I = 17.2 microM, 25 degrees C). In addition, these substitutions lower the reduction potential of the protein and increase the pK(a) for the water molecule coordinated to the heme iron of metmyoglobin. The peroxidase [2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid), ABTS, as substrate] and the Mn(2+)-peroxidase activity of the variant are both increased approximately 3-fold. In contrast to wild-type Mb, both the affinity for azide and the midpoint potential of the variant are significantly influenced by the addition of Mn(2+). The structure of the variant has been determined by x-ray crystallography to define the coordination environment of bound Mn(2+) and Cd(2+). Although slight differences are observed between the geometry of the binding of the two metal ions, both are hexacoordinate, and neither involves coordination by E63. PMID- 12644708 TI - Kinetics of proton-coupled electron-transfer reactions to the manganese-oxo "cubane" complexes containing the Mn4O4(6+) and Mn4O4(7+) core types. AB - The kinetics of proton-coupled electron-transfer (pcet) reactions are reported for Mn(4)O(4)(O(2)PPh(2))(6), 1, and [Mn(4)O(4)(O(2)PPh(2))(6)](+), 1(+), with phenothiazine (pzH). Both pcet reactions form 1H, by H transfer to 1 and by hydride transfer to 1(+). Surprisingly, the rate constants differ by only 25% despite large differences in the formal charges and driving force. The driving force is proportional to the difference in the bond-dissociation energies (BDE >94 kcalmol for homolytic, 1H --> H + 1, vs. approximately 127 kcalmol for heterolytic, 1H --> H(-) + 1(+), dissociation of the OH bond in 1H). The enthalpy and entropy of activation for the homolytic reaction (deltaH = -1.2 kcalmol and deltaS= -32 calmol.K; 25-6.7 degrees C) reveal a low activation barrier and an appreciable entropic penalty in the transition state. The rate-limiting step exhibits no HD kinetic isotope effect (k(H)k(D) = 0.96) for the first H atom transfer step and a small kinetic isotope effect (1.4) for the second step (1H + pzH --> 1H(2) + pz(*)). These lines of evidence indicate that formation of a reactive precursor complex before atom transfer is rate-limiting (conformational gating), and that little or no NH bond cleavage occurs in the transition state. H atom transfer from pzH to alkyl, alkoxyl, and peroxyl radicals reveals that BDEs are not a good predictor of the rates of this reaction. Hydride transfer to 1(+) provides a concrete example of two-electron pcet that is hypothesized for the OH bond cleavage step during catalysis of photosynthetic water oxidation. PMID- 12644707 TI - An FeIV=O complex of a tetradentate tripodal nonheme ligand. AB - The reaction of [Fe(II)(tris(2-pyridylmethyl)amine, TPA)(NCCH(3))(2)](2+) with 1 equiv. peracetic acid in CH(3)CN at -40 degrees C results in the nearly quantitative formation of a pale green intermediate with lambda(max) at 724 nm ( epsilon approximately 300 M(-1).cm(-1)) formulated as [Fe(IV)(O)(TPA)](2+) by a combination of spectroscopic techniques. Its electrospray mass spectrum shows a prominent feature at mz 461, corresponding to the [Fe(IV)(O)(TPA)(ClO(4))](+) ion. The Mossbauer spectra recorded in zero field reveal a doublet with DeltaE(Q) = 0.92(2) mms and delta = 0.01(2) mms; analysis of spectra obtained in strong magnetic fields yields parameters characteristic of S = 1 Fe(IV)O complexes. The presence of an Fe(IV)O unit is also indicated in its Fe K-edge x-ray absorption spectrum by an intense 1-s --> 3-d transition and the requirement for an ON scatterer at 1.67 A to fit the extended x-ray absorption fine structure region. The [Fe(IV)(O)(TPA)](2+) intermediate is stable at -40 degrees C for several days but decays quantitatively on warming to [Fe(2)(mu-O)(mu-OAc)(TPA)(2)](3+). Addition of thioanisole or cyclooctene at -40 degrees C results in the formation of thioanisole oxide (100% yield) or cyclooctene oxide (30% yield), respectively; thus [Fe(IV)(O)(TPA)](2+) is an effective oxygen-atom transfer agent. It is proposed that the Fe(IV)O species derives from OO bond heterolysis of an unobserved Fe(II)(TPA)-acyl peroxide complex. The characterization of [Fe(IV)(O)(TPA)](2+) as having a reactive terminal Fe(IV)O unit in a nonheme ligand environment lends credence to the proposed participation of analogous species in the oxygen activation mechanisms of many mononuclear nonheme iron enzymes. PMID- 12644709 TI - mRNA of placental origin is readily detectable in maternal plasma. AB - The discovery of circulating fetal nucleic acid in maternal plasma has opened up new possibilities for noninvasive prenatal diagnosis. Thus far, a gender- and polymorphism-independent fetal-specific target that can be used for prenatal screening and monitoring in all pregnant women has not been reported. In addition, the origin of such circulating nucleic acid has remained unclear. Here we provide direct evidence that the placenta is an important source of fetal nucleic acid release into maternal plasma by demonstrating that mRNA transcripts from placenta-expressed genes are readily detectable in maternal plasma. The surprising stability of such placental mRNA species in maternal plasma and their rapid clearance after delivery demonstrate that such circulating mRNA molecules are practical markers for clinical use. The measurement of such plasma mRNA markers has provided a gender-independent approach for noninvasive prenatal gene expression profiling and has opened up numerous research and diagnostic possibilities. PMID- 12644710 TI - Structural effects of quinacrine binding in the open channel of the acetylcholine receptor. AB - Noncompetitive inhibitors of the nicotinic acetylcholine (ACh) receptors suppress cation flux directly by binding in and blocking the open channel or indirectly by stabilizing closed states of the receptor. The lidocaine derivative QX-314 and the acridine derivative quinacrine act directly as open channel blockers, but can act indirectly as well. The binding site for quinacrine in the open channel of mouse-muscle ACh receptor was mapped in cysteine-substituted mutants of the alpha subunit expressed with wild-type beta, gamma, and delta subunits. In the open state, substituted cysteines in the inner half of the second membrane-spanning segment (M2), but not in the outer half, were protected by quinacrine from reaction with 2-aminoethyl methanethiosulfonate. In addition, an alkylating derivative, quinacrine mustard, affinity labeled a subset of the substituted cysteines in M2, but only in the open state. These results, mapped onto a model of the open channel surrounded by five alpha-helical M2s, imply that quinacrine binds midway down M2 in the same site previously mapped for QX-314. A cysteine substituted for a residue in the outer third of alphaM1, which reacted with 2 aminoethyl methanethiosulfonate only in the presence of ACh, reacted faster in the additional presence of quinacrine or QX-314. It is proposed that channel opening involves both the opening of the resting gate at the inner end of M2 and the removal of an obstruction formed by the outer end of M1 that retards diffusion of blockers into the closed channel. Blocker binding in the open channel causes a further change in structure. PMID- 12644711 TI - The LIM-only protein FHL2 is a serum-inducible transcriptional coactivator of AP 1. AB - Proteins with LIM domains have been implicated in transcriptional regulation. The four and half LIM domain (FHL) group of LIM-only proteins is composed of five members, some of which have been shown to have intrinsic activation function. Here we show that FHL2 is the only member of the family whose expression is inducible upon serum stimulation in cultured cells. Induction of FHL2 is coordinated in time with the increased levels of two early-response products, the oncoproteins Fos and Jun. FHL2 associates with both Jun and Fos, in vitro and in vivo. The FHL2-Jun interaction requires the Ser-63-Ser-73 JNK phosphoacceptor sites in c-Jun, but not their phosphorylation. FHL2 powerfully stimulates Fos- and Jun-dependent transcription, thereby acting as an inducible coactivator of AP 1 function. Moreover, we show that intracellular localization of FHL2 is controlled by signaling events and a Crm1-dependent active nuclear export mechanism. Thus, FHL2, as an inducible coactivator of AP-1, coordinately participates with Fos and Jun in the early transcriptional response to serum factors. PMID- 12644712 TI - Gapped DNA is anisotropically bent. AB - Ionizing radiation damages DNA in several ways, including through formation of a single-nucleoside gap in one DNA strand. We have developed a two-dimensional gel electrophoresis method to investigate the effect of a strand gap on DNA structure. We generate a library of gapped DNA molecules by treating a DNA restriction fragment with the hydroxyl radical, generated by the reaction of Fe(II) EDTA with hydrogen peroxide. The DNA molecule studied contains a fixed bend produced by a set of phased adenine tracts. The A-tract bend serves as a reference bend for investigating the conformational nature of a strand gap. In the first electrophoretic dimension, a bent DNA molecule that has been treated with the hydroxyl radical is electrophoresed on a native gel. Smearing of the band on the native gel indicates that the library of gapped DNA molecules contains a variety of DNA conformations. In the second electrophoretic dimension, gapped DNA molecules having different native gel mobilities are electrophoresed on separate lanes of a denaturing gel to reveal how each strand gap affects the native gel mobility (and thus shape) of the DNA. Our results demonstrate that a single-nucleoside gap in a DNA duplex leads to an anisotropic, directional bend in the DNA helix axis. The implications of our findings for recognition of this lesion by DNA repair proteins are discussed. PMID- 12644713 TI - In vivo calcium imaging of brain activity in Drosophila by transgenic cameleon expression. AB - Various genetically encoded fluorescent sensors that monitor changes in intracellular calcium concentration have been developed over the last few years. The ability to target these calcium indicators to cells and structures of interest makes them valuable tools for diverse applications and gives them distinct advantages over conventional fluorescent dyes in transgenically tractable organisms. In particular, the cameleon calcium sensors have been used successfully in a number of applications. For example, we use cameleon-2.1 to monitor in vivo brain activity in Drosophila. However, using cameleons to image intracellular calcium concentration changes in vivo is still evolving and is by no means a standard technique. Experimental details and "tricks" for dealing with equipment, techniques, and data evaluation are still restricted to a few laboratories. In this protocol for calcium imaging in Drosophila brain using cameleon-2.1, we provide guidelines to the basic principles of this novel technique in Drosophila neuroscience and, more generally, to the broad field of signal transduction research. PMID- 12644715 TI - Recurrent wheezing in infants and young children and bronchial hyperresponsiveness: a perspective. AB - Epidemiological studies report a 50% incidence of at least one wheezing episode in young children. If we can argue that 10% of children have asthma sometime during their pediatric years, it still leaves a significant percentage of children with an unexplained cause for their wheezing. Other recognized phenotypes of recurrent wheezing include young children exposed to excessive environmental tobacco smoke (ETS), while other infants wheeze recurrently following a significant episode of bronchiolitis. Bronchial hyperresponsiveness (BHR) is a universally recognized phenomenon of asthma, but its presence in young children with recurrent wheezing is not as well studied. Currently available studies demonstrates that BHR is also seen in young pediatric asthmatics, paralleling what is well recognized in adolescent or adult asthma. In those children with post-bronchiolitis wheezing, BHR appears to be present to a degree; while infants and young children exposed to ETS have increased BHR, as a group. If exaggerated BHR in recurrent wheezing children without asthma has the same inherent disadvantage as it does in asthmatic children, additional studies looking directly at this issue in a longitudinal fashion need to be designed. A hypothesis of BHR in non-asthmatic children is presented that could be studied prospectively. PMID- 12644716 TI - Bronchoprovocation methods: direct challenges. AB - Inhalation challenges with direct-acting stimuli histamine and methacholine are widely used to measure airway responsiveness. Three widely used methods (2-min tidal breathing method, breath-activated dosimeter method, hand-held manual nebulizer) are described. Careful standardization is important so as to best differentiate normal from increased airway responsiveness and to permit comparison between methods. With current methods standardized as suggested by the ATS, a methacholine (or histamine) PC(20) > 16 mg/mL is considered normal. A PC(20) < 16 mg/mL is highly sensitive for current symptoms of asthma. Interpretation of methacholine or histamine inhalation test requires that symptoms be current (within a few days) and that FEV(1) be normal. PMID- 12644717 TI - Methods for "indirect" challenge tests including exercise, eucapnic voluntary hyperpnea, and hypertonic aerosols. AB - Bronchial provocation tests that use stimuli that act indirectly to cause airway narrowing have a high specificity for identifying people with active asthma who have the potential to respond to treatment with antiinflammatory drugs. The first test to be developed was exercise and it was used to assess the efficacy of drugs such as sodium cromoglycate. Eucapnic voluntary hyperpnea was developed later, as a surrogate test for exercise. Hypertonic aerosols were introduced to mimic the dehydrating effects of evaporative water loss that occurs during hyperpnea. A wet aerosol of 4.5% saline or a dry powder formulation of mannitol is used. At present the indirect challenge tests are becoming increasingly recognised as appropriate for monitoring treatment with inhaled steroids. Indirect tests identify those with potential for exercise-induced bronchoconstriction, an important problem for some occupations, such as the defence forces, fire fighters and the police force and for some athletic activities. The advantage in using an indirect challenges, over a direct challenge with a single pharmacological agonist, is that a positive response indicates that inflammatory cells and their mediators (prostaglandins, leukotrienes and histamine) are present in the airways in sufficient numbers and concentration to indicate that asthma is active at the time of testing. The corollary to this is that a negative test in a known asthmatic indicates good control or mild disease. Another advantage is that healthy subjects do not have significant airway narrowing to indirect challenge tests. The protocols used for challenge with indirectly acting stimuli are presented in detail. PMID- 12644718 TI - Bronchodilation and bronchoprotection by deep inspiration and their relationship to bronchial hyperresponsiveness. AB - Bronchial hyperresponsiveness (BHR) is a cardinal feature of asthma. Airway inflammation and BHR are probably linked, but the mechanisms underlying this relationship remain elusive. BHR is closely associated with defects in the beneficial responses to lung inflation. These responses, which become apparent by the fact that healthy individuals can develop severe airway obstruction if they are exposed to methacholine in the absence of deep inspirations, include bronchodilation and bronchoprotection. Bronchodilation refers to the effect of lung inflation after the induction of airway smooth muscle tone, while bronchoprotection is used to indicate the effect prior to inhalation of a spasmogen. Mild asthmatics who manifest BHR lack bronchoprotection by lung inflation. In contrast, many of them are able to bronchodilate. In more severe disease, both functions are impaired. The lack of bronchoprotection is also found in individuals with rhinitis and BHR, but no asthma. These and other observations suggest that the mechanisms of bronchodilation and bronchoprotection may be distinct, although overlap is possible. We believe that the loss of bronchoprotection is pertinent to the phenomenon of hyperresponsiveness, but that both the bronchodilatory and bronchoprotective functions of deep inspiration interact to produce the asthmatic phenotype. In this review, we describe the phenomena of lung inflation-induced bronchodilation and bronchoprotection and detail potential mechanical and neurohumoral mechanisms accounting for these physiologic functions. In addition, possible mechanisms leading to the impairment of these functions in subjects with BHR are discussed. PMID- 12644719 TI - What evidence implicates airway smooth muscle in the cause of BHR? AB - Bronchial hyperresponsiveness (BHR), the occurrence of excessive bronchoconstriction in response to relatively small constrictor stimuli, is a cardinal feature of asthma. Here, we consider the role that airway smooth muscle might play in the generation of BHR. The weight of evidence suggests that smooth muscle isolated from asthmatic tissues exhibits normal sensitivity to constrictor agonists when studied during isometric contraction, but the increased muscle mass within asthmatic airways might generate more total force than the lesser amount of muscle found in normal bronchi. Another salient difference between asthmatic and normal individuals lies in the effect of deep inhalation (DI) on bronchoconstriction. DI often substantially reverses induced bronchoconstriction in normals, while it often has much less effect on spontaneous or induced bronchoconstriction in asthmatics. It has been proposed that abnormal dynamic aspects of airway smooth muscle contraction velocity of contraction or plasticity elasticity balance might underlie the abnormal DI response in asthma. We suggest a speculative model in which abnormally long actin filaments might account for abnormally increased elasticity of contracted airway smooth muscle. PMID- 12644721 TI - Inhibitors of actin filament polymerisation attenuate force but not global intracellular calcium in isolated pressurised resistance arteries. AB - Receptor-coupled contractile activation of arterial smooth muscle involves increases in intracellular calcium ([Ca(2+)](i)) and subsequent alteration of myosin light chain phosphorylation. An additional mechanism whereby agonists could regulate vascular contractility may be alteration of actin filament dynamics. Therefore, in this study, we have investigated the influence of two inhibitors of actin filament polymerisation, cytochalasin D and latrunculin B, on the [Ca(2+)](i) and force responsiveness of pressurised rat mesenteric arteries to alpha-adrenergic stimulation. Following cytochalasin D or latrunculin B treatment, phenylephrine-induced constrictions were significantly reduced to 11 +/- 3.2% (n = 6) and 10 +/- 4.4% (n = 6) of control, respectively, whereas [Ca(2+)](i) remained at 98 +/- 21% and 104 +/- 7.0% of control, respectively. Such effects of cytochalasin D were not restricted to mesenteric small arteries. Cytochalasin D also significantly reduced the force, but not [Ca(2+)](i) responses to agonist stimulation in other vascular (portal vein) and non-vascular (uterine) tissues. These data indicate that inhibitors of net actin polymerisation attenuate maximum agonist-induced force responsiveness without similar reductions in [Ca(2+)](i) in pressurised resistance vessels and other smooth muscle tissues. This suggests that modulation of the dynamic equilibrium between filamentous F-actin and monomeric globular actin (G-actin) may be an important mechanism, acting independently of global [Ca(2+)](i) homeostasis, to regulate the smooth muscle contractile state. PMID- 12644720 TI - Airway hyperresponsiveness: a story of mice and men and cytokines. AB - Bronchial hyperresponsiveness (BHR) is an essential part of the definition of asthma. Although our understanding of the allergic inflammatory and immunologic mechanisms of asthma have markedly increased, the mechanism of BHR remains to be elucidated. Increased BHR is associated temporally with exposure to allergens, certain respiratory viruses, pollutants such as ozone, and certain occupational chemicals. An important research use of determining the degree of BHR to direct and indirect challenge is to determine the efficacy of pharmacologic and immunodulatory agents. Beta-adrenergic agents inhibit BHR and certain genetic polymorphisms of the beta-adrenergic receptor are associated with increased BHR. When beta-adrenergic receptors are blocked, sensitivity to allergens is markedly increased in patients with asthma and animal models of asthma. Allergen challenge and clinical asthma are associated with synthesis and release of pro-inflammatory cytokines such as IL-1 and TNF-alpha which have been shown to decrease the response to beta-agonists and increased the reactivity to methacholine and the airways neutrophils and alveolar macrophages. The Th2 cytokine IL-13 is increased in the airways of asthmatics and increases BHR in normal unsensitized animals. The mechanisms of this effect of IL-13 are being intensively investigated. Our group has shown that IL-13 induced BHR persisted for at least 7 days and the soluble receptor IL-13R2alpha protected against their BHR. Other investigators have demonstrated that IL-13 is necessary and sufficient for the induction of BHR and that eosinophilic airway inflammation in the absence of IL-13 fails to induce BHR. These studies indicate that treatment of human asthma with antagonists of IL 13 may be very effective. PMID- 12644722 TI - Intercellular electrical coupling in vascular cells present in rat intact cerebral arterioles. AB - Using arterioles dissected from rat cerebral pial membrane, we investigated cell to-cell communication by means of the amphotericin-B-perforated whole-cell patch clamp technique. During a brief voltage step or ramp in voltage clamp mode, the leak current through the membrane of a smooth muscle cell in situ in an arteriole was attenuated by different types of gap junction inhibitors. Macroscopic current recording via two electrodes attached to adjacent smooth muscle cells or to a smooth muscle cell and a nearby endothelial cell revealed transjunctional current flow between cells of the same or different types. The pseudo-junctional conductance-voltage relationship obtained from adjacent smooth muscle cells differed from that obtained from a smooth muscle cell and an endothelial cell (symmetrical for the former and asymmetrical for the latter). Single- or multiple channel events could be recorded from weakly coupled cells of the two types: simultaneous, equal-sized events in opposite directions were recorded via two electrodes, demonstrating electrical coupling directly between cell pairs. The single-channel conductance observed most frequently varied: 200-230 pS for pairs of two smooth muscle cells and 240-260 pS for mixed pairs. These data suggest the existence of differential patterns of electrical communication in vascular cells present in rat intact cerebral arterioles. PMID- 12644723 TI - Cyclic ADP-ribose contributes to contraction and Ca2+ release by M1 muscarinic receptor activation in coronary arterial smooth muscle. AB - The present study determined the role of cyclic ADP-ribose (cADPR) in mediating vasoconstriction and Ca(2+) release in response to the activation of muscarinic receptors. Endothelium-denuded small bovine coronary arteries were microperfused under transmural pressure of 60 mm Hg. Both acetylcholine (ACh; 1 nmol/L to 1 micromol/L) and oxotremorine (OXO; 2.5-80 micromol/L) produced a concentration dependent contraction. The vasoconstrictor responses to both ACh and OXO were significantly attenuated by nicotinamide (Nicot; an ADP-ribosyl cyclase inhibitor), 8-bromo-cADPR (8-Br-cADPR; a cADPR antagonist) or ryanodine (Ry; an Ry receptor antagonist). Intracellular Ca(2+) ([Ca(2+)](i)) was determined by fluorescence spectrometry using fura-2 as a fluorescence indicator. OXO produced a rapid increase in [Ca(2+)](i) in freshly isolated single coronary arterial smooth muscle cells (CASMCs) bathed with Ca(2+)-free Hanks' solution. This OXO induced rise in [Ca(2+)](i) was significantly reduced by pirenzepine (PIR; an M(1) receptor-specific blocker), Nicot, 8-Br-cADPR or Ry. The effects of OXO on the activity of ADP-ribosyl cyclase (cADPR synthase) were examined in cultured CASMCs by measuring the rate of cyclic GDP- ribose (cGDPR) formation from beta nicotinamide guanine dinucleotide. It was found that OXO produced a concentration dependent increase in the production of cGDPR. The stimulatory effect of OXO on ADP-ribosyl cyclase was inhibited by both PIR and Nicot. These results suggest that the cADPR signaling pathway participates in the contraction of small coronary arterial smooth muscle and Ca(2+) release induced by activation of M(1) muscarinic receptors. PMID- 12644724 TI - Transforming growth factor-beta-dependent events in vascular remodeling following arterial injury. AB - Constrictive remodeling has been identified as a major contributor to restenosis following angioplasty. Characterization of transforming growth factor-beta (TGF beta)-mediated cellular events in the adventitia and their contribution to vascular remodeling, however, has not previously been studied in detail. The balloon catheter denudation model was performed on rat carotid artery, and groups of rats were treated with vehicle or a TGF-beta inhibitor, a soluble TGF-beta receptor type II (TGF-beta R:Fc). Adventitial cell proliferation, which peaked 4 days after injury, was characterized by the de novo formation of several cell layers surrounding the outer adventitia and this process was not dependent upon TGF-beta activity. These neoadventitial cells expressed an abundance of collagen type I and a fetal isoform of fibronectin containing the EIIIA domain, and the expression of both proteins was suppressed in the presence of TGF-beta R:Fc. Lumenal narrowing was apparent 14 days after injury. Inhibition of TGF-beta signaling promoted vessel enlargement. As a result, lumen size did not change despite neointima formation. In conclusion, adventitial fibrosis with abundant collagen matrix deposition but not adventitial cell proliferation is dependent upon endogenous TGF-beta activity. Furthermore, inhibition of TGF-beta signaling prevents injury-induced reduction in lumen area by promoting vessel enlargement. PMID- 12644725 TI - Lack of nitric oxide mediation of flow-dependent arteriolar dilation in type I diabetes is restored by sepiapterin. AB - The mechanisms leading to microangiopathy in diabetes mellitus have still not been clearly elucidated. We hypothesized that type I diabetes mellitus affects the endothelium and alters flow-dependent dilation of arterioles, an important mechanism involved in local regulation of blood flow. Isolated, pressurized gracilis muscle arterioles (inside diameter approximately 150 microm at 80 mm Hg) from rats with streptozotocin (STZ)-induced diabetes mellitus exhibited reduced dilations induced by increases in perfusate flow compared to those of normal rats (plasma glucose: 25.7 +/- 0.7 vs. 6.4 +/- 0.5 mmol/l; maximum increase in diameter: 15 +/- 4 vs. 31+/- 3 microm, p < 0.05). In control arterioles, both nitric oxide (NO) and prostaglandins mediated the flow-dependent dilation, whereas flow-induced dilations of diabetic arterioles were unaffected by N(omega) nitro-L-arginine methyl ester (L-NAME) and were abolished by indomethacin. Sepiapterin - precursor of the endothelial NO synthase (eNOS) cofactor tetrahydrobiopterin (BH(4)) - restored the L-NAME-sensitive portion of flow dependent dilations of diabetic arterioles. Furthermore, depletion of BH(4) by 2,4-diamino-6-hydroxypyrimidine (DAHP) in control arterioles also resulted in reduced flow-dependent dilations, which were restored by intraluminal sepiapterin [but not with superoxide dismutase (SOD) plus catalase (CAT) (SOD+CAT)] and then could be inhibited by L-NAME. Dilations induced by the NO donor sodium nitroprusside (SNP) were unaffected by L-NAME in diabetes mellitus arterioles or when eNOS was activated by intraluminal flow in DAHP-treated arterioles (with or without SOD+CAT). In contrast, pyrogallol (known to produce reactive oxygen species) substantially reduced acetylcholine- and SNP-induced dilation in a SOD+CAT-reversible manner. Collectively, these findings suggest that in diabetic arterioles, due to the reduced bioavailability of BH(4), the synthesis of NO by eNOS is limited, resulting in a reduced flow-induced dilation, a mechanism that may also be responsible for the development of diabetic microangiopathy and exacerbation of other vascular diseases. PMID- 12644726 TI - Blunted coronary vasoreactivity to insulin is an early alteration in hypertension. AB - Insulin resistance in the heart is not localized to the myocardium, but may also occur in blood vessels. We studied the effects of insulin on coronary vasodilation in hypertension. Coronary vascular resistance was quantitated in 11 nonsmoking men with untreated mild essential hypertension and 9 healthy normotensive men using positron emission tomography and (15)O-labeled water. The measurements were performed at baseline and during adenosine infusion (140 microg x kg(-1) x min(-1)) with or without simultaneous euglycemic physiological (serum insulin approximately 70 mU/l) and supraphysiological (serum insulin approximately 460 mU/l) hyperinsulinemia. Coronary resistance was significantly higher in hypertensive than normotensive subjects at baseline and during adenosine infusion. Physiological hyperinsulinemia decreased hyperemic coronary resistance significantly in both groups. Supraphysiological hyperinsulinemia further decreased the hyperemic coronary resistance in normotensive but not in hypertensive subjects, leading to higher hyperemic coronary resistance in hypertensive than normotensive subjects (27.2 +/- 8.7 vs. 19.2 +/- 4.9 mm Hg x min x g x ml(-1), p < 0.05). However, insulin-stimulated whole body glucose uptake values were similar between the groups during both insulin infusions. In conclusion, insulin-induced coronary vasodilation is blunted in young subjects with mild essential hypertension who are otherwise healthy. Coronary vascular resistance to insulin occurs although no change in peripheral glucose uptake can be detected. While we do not know whether the same results can be extrapolated to female or older subjects, these results indicate a novel defect in the regulation of coronary arteries in the early phase of hypertension. PMID- 12644727 TI - The roles of nitric oxide in dilating proximal and terminal arterioles of skeletal muscle during systemic hypoxia. AB - Indirect evidence suggests that dilatation evoked by systemic hypoxia in proximal and terminal arterioles of skeletal muscle is mediated by adenosine acting on A(1) receptors, but that the dilatation of proximal arterioles requires the presence of nitric oxide (NO), whereas that of terminal arterioles is mediated by NO. In the present study, we showed that primary and terminal arterioles of spinotrapezius muscle (diameters: 45 +/- 4 and 9 +/- 1 microm, respectively) in anaesthetised rats dilated by 15 +/- 7 and 48 +/- 24%, respectively, during hypoxia (breathing 12% O(2)). Inhibition of NO synthesis with intravenous nitro-L arginine methyl ester attenuated these responses (to -2 +/- 3 and -4 +/- 4%, respectively). However, when a tonic level of NO was subsequently restored by infusion of NO donor, hypoxia evoked dilatation of primary arterioles (+24 +/- 8%) but not terminal arterioles (0 +/- 4%). The adenosine A(1) receptor antagonist DPCPX (8-cyclopentyl-1,3-dipropylxanthine) (intravenous) attenuated this 'restored' primary arteriolar vasodilatation (to +6 +/- 3%). Similar results were obtained with 8% O(2). We propose that (1) proximal arteriolar dilatation evoked by systemic hypoxia depends on a background level of NO, which facilitates the action of adenosine on A(1) receptors, and (2) hypoxia-evoked dilatation of terminal arterioles is mediated by NO synthesised via A(1) receptor stimulation. PMID- 12644728 TI - Isoprostane 8-epi-prostaglandin F2 alpha decreases lymph capillary pressure in patients with primary lymphedema. AB - In patients with lymphedema, reduced lymph drainage capacity results in an overloaded superficial microlymphatic network and microlymphatic hypertension. In in vitro experiments, it has been shown that 8-epi-prostaglandin F2 alpha (PGF) induced contractions in human lymphatics. Since lymphatic contractility plays a crucial role in the regulation and generation of lymph transport, we studied the effect of PGF on microlymphatic dynamics by measuring lymph capillary pressure (LCP). Twenty healthy volunteers and 13 patients with primary lymphedema were studied after either PGF or placebo was applied to the skin and occlusively covered for 30 min. Glass micropipettes (7-9 microm) were inserted under microscopic control into initial lymphatics visualized by fluorescence microlymphography and pressure measurements were performed using the servo nulling technique. The mean LCP in patients with lymphedema was significantly higher (19.8 +/- 12.1 mm Hg) than that in healthy controls (8.4 +/- 4.1 mm Hg) at the placebo-treated site and decreased to normal values after PGF (10.0 +/- 7.7 mm Hg). In healthy volunteers, there was no significant decrease of LCP with PGF compared to placebo. PGF normalizes microlymphatic hypertension in patients with lymphedema by improving lymph transport into deeper channels. PMID- 12644729 TI - Role of pRb family members in injured carotid arteries. PMID- 12644730 TI - Experiences with an outpatient relapse program (community reinforcement approach) combined with naltrexone in the treatment of opioid-dependence: effect on addictive behaviors and the predictive value of psychiatric comorbidity. AB - BACKGROUND: There is increasing interest in naltrexone, an opiate antagonist, in the treatment of opiate addicts. The effects of naltrexone are often compromised by a lack of compliance and drop-out. The effects of this compound are probably more favorable when combined with a psychosocial intervention such as the Community Reinforcement Approach (CRA). AIM: To explore the effects of a combination therapy (naltrexone plus CRA treatment) and the predictive value of sociodemographic and psychopathologic characteristics. METHOD: Using a before-and after design, heroine addicts (n = 24) receiving a combined naltrexone plus CRA treatment are compared with a group (n = 20) on methadone maintenance therapy (reference group). RESULTS: Over a period of 6 months, 58% (14/24) did not relapse, after 1 year at least 55% (12/22) still met the initial goal of continuous abstinence. At baseline, the treatment group and the reference group were similar on nearly all variables except for the number of times clients were arrested. Within the treatment group, a comparison was made between the continuous abstinent and those who relapsed into frequent opioid use. Differences were significant in the cluster-B personality disorders and in polydrug users. CONCLUSION: The combination of naltrexone plus intensive CRA in an outpatient setting appears to be promising. A high score on cluster-B and polydrug use is associated with relapse. PMID- 12644731 TI - Cost-effectiveness of adjuvant treatment with acamprosate in maintaining abstinence in alcohol dependent patients. AB - An open prospective cohort study was performed in Germany in order to evaluate the costs of treating alcohol dependence under real-world conditions. Eight hundred and fourteen recently detoxified alcohol-dependent patients were provided with psychosocial rehabilitation support. In addition, 540 alcohol-dependent patients treated with adjuvant acamprosate therapy were compared with 274 patients without pharmacotherapy. Real costs were assessed over a period of one year. Of the patients who were treated with acamprosate, 33.6% remained abstinent compared to only 21.1% in the standard cohort. The mean total costs per patient treated with acamprosate were EUR 1,631.49 per year. In the standard cohort, total costs were EUR 2,068.83. This difference is highly significant (p = 0.012). Direct costs amounted to 76.9% of the total costs, with a 27% difference between the cohorts (p < 0.001). There was no difference in indirect costs between the two groups (p = 0.324). This real-cost study confirms the favourable cost effectiveness of acamprosate previously suggested by pharmaco-economic modelling studies. PMID- 12644732 TI - Flupenthixol in relapse prevention in schizophrenics with comorbid alcoholism: results from an open clinical study. AB - Substance use, especially alcoholism, has been recognized as a significant problem in schizophrenic patients, though only a few studies on the effects of pharmacotherapy in these patients have been conducted so far. The thioxanthene neuroleptic flupenthixol, which can be given intramuscularly (i.m.) for improving compliance, has been studied as a possible anti-craving drug both in animal models of alcoholism and some clinical studies. Pilot studies suggest that comorbid schizophrenics with substance use may benefit from treatment with flupenthixol. Efficacy of flupenthixol (10-60 mg i.m.) in reducing alcohol consumption of dual diagnosis patients was studied in an open 6-month clinical trial in 27 schizophrenics with comorbid alcoholism. Twenty-one patients entered the intention-to-treat analysis. Fourteen subjects were completers, 13 dropped out. Six patients completely abstained from alcohol during treatment. Alcohol consumption was significantly reduced compared to baseline (4 weeks before treatment as measured by timeline follow-back interview). In general, while patients showed a marked improvement concerning alcohol consumption, only a slight improvement in psychopathology was recorded. Overall tolerability was good. These data indicate a probable beneficial effect of flupenthixol in schizophrenic patients with comorbid alcoholism. Although the efficacy of flupenthixol as an anti-craving drug in dual diagnosis patients has to be explored in further studies, the drug may be considered a promising medication for these patients. PMID- 12644733 TI - Evaluating long-term effects of heroin-assisted treatment: the results of a 6 year follow-up. AB - Since January 1994, heroin-assisted treatment for opiate addicts has been available in Switzerland. This is the first report of the long-term effects of this form of treatment. The report examines subjects who entered a study involving medical prescription of opiates (Projekt zur arztlichen Verschreibung von Betaubungsmitteln; PROVE) in Switzerland between January 1994 and March 1995 (n = 366). Opiates were dispensed in eight treatment centres. A follow-up was conducted 6 years after treatment entry. Two groups were assessed: clients who have continuously been on heroin-assisted treatment since entry into the PROVE study or who re-entered this treatment, and ex-clients who had discontinued heroin-assisted treatment at the time of follow-up. Two kinds of comparisons were conducted. Firstly, conditions at treatment entry were compared to 6-year follow up outcomes, and secondly, outcomes were compared between clients still on heroin assisted treatment and those who had been discharged. It was found that 46% of the clients still alive were on heroin-assisted treatment at the time of follow up. A comparison of the present living conditions showed very little difference between those in treatment and those who had terminated treatment. Compared to the situation at entry, the results of the follow-up showed a significant decrease in the use of illegal substances, illegal income and most other variables concerning social conditions, but they also showed an increase in unemployment and reliance on social benefits. Heroin-assisted treatment is thus efficacious in the long-term course of treatment and is still effective after termination of treatment with respect to living conditions and use of illicit substances. PMID- 12644734 TI - A 25-year follow-up study of drug addicts hospitalised for acute hepatitis: present and past morbidity. AB - The aim of the study was to investigate present and past morbidity in drug addicts, 25 years after hospitalisation for acute hepatitis B or hepatitis nonA nonB. The hospital records for 214 consecutively admitted patients were analysed, and a follow-up study on 66 of the 144 patients still alive was performed. At follow-up, 1 of 54 (1.8%) hepatitis B patients was still HBsAg positive. Twelve patients originally diagnosed as hepatitis nonA-nonB were all among 54 found to be anti-hepatitis C virus (anti-HCV) positive, and the total anti-HCV prevalence was 81.8%. Twelve (22.2%) of the HCV cases were unknown before the follow-up examination. Four (6.1%) participants were anti-human immunodeficiency virus positive, only 1 was on antiretroviral therapy, and none had developed AIDS. Other chronic somatic diseases were a minor problem, whereas drug users reported skin infections as a frequent complication. Forty-three patients (65%) had abandoned addictive drugs since the hospital stay. Serious mental disorders were reported by 19 patients (28.8%), and 17 (25.8%) regarded themselves as present (9) and former (8) compulsive alcohol drinkers. A large proportion of the participants were granted disability pension (39%), a majority because of psychiatric disorders, drug and alcohol abuse. PMID- 12644735 TI - A new measure of the alcohol dependence syndrome: the severity scale of alcohol dependence. AB - The purpose was to develop a diagnostic instrument of the Alcohol Dependence Syndrome defined by Edwards and Gross (1976) since comparable scales show some limitations. The methods include three studies carried out in order to develop the SEverity Scale of Alcohol dependence (SESA; total n = 774 alcohol-dependent patients), and three further studies served for validation (total n = 603 alcohol dependents). The findings reveal that the SESA, a 33-item questionnaire including 7 subscales which cover the criteria of the Alcohol Dependence Syndrome, proved to be internally consistent. The subscales are correlated with respective information from internationally validated interview instruments for the diagnosis of alcohol dependence according to DSM and ICD (CIDI, SCAN): six subscales are correlated with six of eight CIDI items and with eight of nine SCAN items selected for the purpose of validation. In addition, there are positive correlations with the Munich Alcoholism Test, the Michigan Alcoholism Test as well as single items of the WHO Alcohol Use Disorders Identification Test. It is concluded from the results that the SESA shows good validity. PMID- 12644736 TI - Drug consumption rooms in rotterdam: an explorative description. AB - The Rotterdam Drug Monitoring System used survey data, fieldnotes and interviews with staff to investigate the functioning of four (out of six) consumption rooms in Rotterdam. The results show that for most drug users, access to the drug consumption room results in less frequent drug use in public places and more time and rest. Pass holders value being able to use drugs safely inside, and make use of the additional services provided, such as refreshments, washing/showering facilities and talking with others about their personal problems. Two 'weak points' reported by the drug users are discussed in relation to their personal health situation and public nuisance reduction. PMID- 12644737 TI - Progress towards understanding the neurobiology of Batten disease or neuronal ceroid lipofuscinosis. AB - PURPOSE OF REVIEW: The identification of genes mutated in the neuronal ceroid lipofuscinoses has accelerated research into the mechanisms that underlie these fatal autosomal recessive storage disorders, which are often referred to as Batten disease. This review summarizes progress in this field since October 2001, describing advances in cell biology, the characterization of new animal models of neuronal ceroid lipofuscinosis, and the impact of novel methodology to reveal insights into its pathogenesis. RECENT FINDINGS: Gene products for six of the eight forms of neuronal ceroid lipofuscinosis have now been discovered, and concerted efforts are underway to understand the normal biology of each gene product and how this may be altered by mutation. Several lines of evidence point to functions for the CLN genes in the endosomal-lysosomal system, and suggest neuron-specific roles for these proteins. Indeed, a requirement for appropriate protein trafficking within neurons may explain the profound and selective effects of these disorders upon the central nervous system. The development of mouse and large animal models has enabled comparative studies of the progressive effects of disease, including characterization by morphological and biochemical means supplemented by metabonomic and microarray techniques. SUMMARY: Insights into disease mechanisms are building a detailed profile of the impact of neuronal ceroid lipofuscinosis upon the brain. With the eventual aim of developing successful therapeutic strategies, it will be equally important to characterize the clinical progression of the disorder, and to identify quantifiable endpoints that can ultimately be used in clinical trials. PMID- 12644738 TI - Radiation injury and neurogenesis. AB - PURPOSE OF REVIEW: For many cancers, survival depends on aggressive combined therapies, but treatment comes at a price. Children and adults who receive radiotherapy involving the brain frequently experience a progressive cognitive decline. The overt pathologies of radiation injury such as white matter necrosis or vasculopathy are the obvious "smoking guns" of dysfunction. However, many patients exhibit severe learning and memory deficits with no overt pathologic changes. This is especially true when the radiation field involves the temporal lobes. The cause of this debilitating dysfunction is currently unknown and untreatable. RECENT FINDINGS: Within the temporal lobe, the hippocampal formation plays a central role in short-term learning and memory--the functions most notably affected by radiation. Recent work has also shown that hippocampus dependent learning and memory are strongly influenced by the activity of neural stem cells and their proliferative progeny. The hippocampal granule cell layer undergoes continuous renewal and restructuring by the addition of new neurons. Radiation at much lower doses than that needed to injure the more resistant post mitotic neurons and glia of the brain has been found to affect these highly proliferative progenitors severely. The stem/progenitor cell is so sensitive to radiation that a single low dose to the cranium of a mature rat is sufficient to ablate hippocampal neurogenesis. SUMMARY: Progressive learning and memory deficits following irradiation may be caused by the accumulating hippocampal dysfunction that results from a long-term absence of normal stem/progenitor activity. Here, the authors describe the nature of this stem cell dysfunction and contemplate how restoration of stem/progenitor cell activity might be approached in experimental models and, eventually, the clinic. PMID- 12644740 TI - Neuroimaging in developmental disorders. AB - PURPOSE OF REVIEW: This review considers the role of neuroimaging in developmental disorders by highlighting recent studies in two distinct, but overlapping, developmental disorders: autism and fragile X syndrome. RECENT FINDINGS: After a decade of conflicting results in neuroimaging studies of autism, recent studies have provided some convergent data. One well-replicated finding is that autistic subjects have larger brains. Further, this enlargement, present as early as 3 years of age, appears to represent accelerated growth in infancy and may be followed by slowed growth in late childhood. Other findings are discussed but considered preliminary in the absence of converging evidence or replication studies. Recent work in fragile X syndrome suggests aberrant fronto striatal and fronto-parietal networks and relates these abnormalities "forward" to behavior and "backward" to decreased protein expression. SUMMARY: As the field of neuroimaging has matured, it has revealed its promise as a safe, reliable, in vivo tool in the study of developmental disorders. By insisting on larger, more homogeneous patient groups and longitudinal rather than cross-sectional studies, the field is poised to fulfill its ultimate role of linking defects in molecular biology to aberrant behavior. PMID- 12644739 TI - Mouse models of holoprosencephaly. AB - PURPOSE OF REVIEW: Holoprosencephaly (HPE) is the most common anomaly of forebrain development in humans. The pathogenesis of HPE results in a failure of the brain hemispheres to separate during early development. Here we review experimental models of HPE in which some of the genes known to cause HPE in humans have been disrupted in the mouse. RECENT FINDINGS: To date, mutations that cause HPE have been identified in seven genes. Three of these genes encode members of the Sonic hedgehog (SHH) signaling pathway, which regulates the development of ventral structures throughout the neuraxis. Two other HPE mutations affect signaling by Nodal ligands, which also play important roles in neural patterning. The roles of the two other known HPE genes are not yet clear. Analysis of genetically altered mice has revealed that mutations in other members of the SHH and Nodal signaling pathways also result in HPE phenotypes. SUMMARY: Studies of HPE in the mouse have provided a framework for understanding key developmental events in human brain development and may provide new candidate genes for human HPE. Despite this progress, fundamental mysteries remain about how molecules that pattern ventral brain regions ultimately disrupt the formation of the cerebral hemispheres in dorsal regions. PMID- 12644741 TI - The injury response in the term newborn brain: can we neuroprotect? AB - PURPOSE OF REVIEW: Perinatal hypoxia-ischemia is responsible for significant morbidity and mortality in term infants. The developing brain is uniquely vulnerable to hypoxic-ischemic injury, with a complex evolution of injury that affords opportunities for intervention, yet potentially jeopardizes normal maturational processes. RECENT FINDINGS: Data published over the past year have provided insights into the evolution of injury, and have suggested a greater role for oxidants and inflammatory mediators. SUMMARY: The search for ideal neuroprotective agents and techniques for timely injury detection is actively progressing and has led to advances in our understanding of pathogenesis. PMID- 12644742 TI - Understanding Niemann-Pick type C disease: a fat problem. AB - PURPOSE OF REVIEW: Progressive neurological deterioration is the ultimate cause of premature death in Niemann-Pick type C disease. Yet it remains unknown why a defect in basic cellular lipid homeostasis would lead to such profound neurological dysfunction and degeneration. The established belief that the central nervous system disorder of Niemann-Pick type C disease is secondary to lipid accumulation has led to a deficiency of research and information on the neurological manifestation of the disease. Lipid and vesicular trafficking have been studied extensively in non-neuronal cells such as fibroblasts and significant advances have been summarized in excellent reviews. The purpose of the present review is to consolidate the findings on brain lipid metabolism and Niemann-Pick type C disease neuropathology, so as to generate a current picture of the central nervous system disease, and emphasize potential aspects for future research. RECENT FINDINGS: Some key findings in the last couple of years have brought the neurological features of Niemann-Pick type C disease into focus. Most importantly, multiple strategies for reducing lipid overload have proven ineffective in preventing the onslaught of neurodegeneration, while efficiently reducing lipid disturbances in extracerebral tissues. Of much significance was the finding that the central nervous system disease is brain autonomous. Finally, investigation of lipid defects and neuropathological changes in Niemann-Pick type C disease gene-1-deficient mice has shed light on some novel putative therapeutic targets. SUMMARY: The independence of brain pathology from visceral complications in Niemann-Pick type C disease has implications for its treatment. Further studies of the neurological mechanisms underlying the disease will have a significant impact on future clinical diagnosis and management of patients. PMID- 12644743 TI - The current status of neuroimaging for epilepsy: editorial review. PMID- 12644744 TI - The epidemiology of epilepsy revisited. AB - PURPOSE OF REVIEW: Epidemiology is the study of the dynamics of a medical condition in a population. There are many shortcomings in the understanding of the epidemiology of epilepsy mostly caused by methodological problems. These include diagnostic accuracy, case ascertainment, and selection bias. In this article recent progress in this area is discussed and suggestions for future research are made. RECENT FINDINGS: It is generally accepted that in developed countries the incidence is around 50/100 000/year. In resource-poor countries, the incidence is likely to be higher. Prevalence of active epilepsy is in the range of 5-10/1000 in most locations, although it might be higher in some isolates. Age-specific incidence rates have changed, with a decrease in younger age groups and an increase in persons above 60 years. The overall prognosis for seizure control is good and over 70% will enter remission. Epilepsy carries an increased risk of premature death particularly in patients with chronic epilepsy. Sudden unexpected death has been increasingly recognized as a major culprit for this increased mortality. SUMMARY: There is geographic variation in the incidence of epileptic syndromes likely to be associated with genetic and environmental factors, although as yet causality has not been fully established. The complete range of aetiologies in the general population is not known. Few predictors of outcome are recognized and it is difficult to prognosticate in any individual case. Knowledge is patchy about the epidemiology of sudden unexpected death in epilepsy. Future epidemiological research needs to address these issues if we are to progress. PMID- 12644745 TI - Channelopathies as a genetic cause of epilepsy. AB - PURPOSE OF REVIEW: This review describes the significant number of new gene associations with epilepsy syndromes that have emerged during the past year, together with additional mutations and new electrophysiological data relating to previously known gene associations. RECENT FINDINGS: Autosomal dominant juvenile myoclonic epilepsy was demonstrated to be a channelopathy associated with a GABA(A) receptor, alpha1 subunit mutation. Benign familial neonatal infantile seizures were delineated as another channelopathy of infancy, by molecular characterization of sodium channel, alpha2 subunit defects. A sodium channel, alpha2 subunit defect was previously found to be associated with generalized epilepsy with febrile seizures plus. Similarly, the clinical spectrum associated with potassium channel, KQT-like mutations was extended to include the channelopathy myokymia and neonatal epilepsy. Mutations in the non-ion channel genes, leucine-rich, glioma inactivated 1 gene and Aristaless related homeobox gene, have emerged as important causes of their specific syndromes, with mutations in the latter gene frequently underlying X-linked mental retardation with epilepsy. SUMMARY: All but one of the idiopathic epilepsies with a known molecular basis are channelopathies. Where the ion channel defects have been identified, however, they generally account for a minority of families and sporadic cases with the syndrome in question. The data suggest that ion channel mutations of large effect are a common cause of rare monogenic idiopathic epilepsies, but are rare causes of common epilepsies. Additive effects of genetic variation, perhaps within the same ion channel gene families, are likely to underlie the common idiopathic generalized epilepsies with complex inheritance. The genetics of epilepsy is progressing rapidly toward a more detailed molecular dissection and definition of syndromes. PMID- 12644746 TI - Neonatal seizures and their treatment. AB - PURPOSE OF REVIEW: Neonatal seizures continue to present a diagnostic and therapeutic challenge to paediatricians worldwide, and are a worrying sign for both parents and clinicians alike. The present review summarizes recent evidence regarding the diagnosis, aetiology and treatment of neonatal seizures. It is timely because there is new evidence that seizures are damaging to the neonatal brain, and because prolonged electroencephalographic recordings during treatment have provided information that challenges established treatment regimens. RECENT FINDINGS: Neonatal seizures can permanently disrupt neuronal development, induce synaptic reorganization, alter plasticity and "prime" the brain to increased damage from seizures later in life. Phenobarbitone remains the mainstay of treatment and is effective in about one-third of cases; babies who respond tend to have a smaller seizure burden and a relatively normal background electroencephalogram. Their prognosis is better than in those who require second line treatments. Phenytoin and lignocaine (membrane stabilizing drugs) are probably more effective than any of the benzodiazepines as second line, but very few evaluation studies have been reported. Babies who require second-line treatments are more likely to have hypoxic ischaemic encephalopathy, an abnormal background electroencephalogram and a large seizure burden, and have a worse prognosis than do those who respond to a single agent; most have significant disability at follow up. SUMMARY: The search for an effective antiepileptic regimen in the newborn must continue. Whether better control of neonatal seizures leads to a reduction in neurodisability in childhood cannot be determined until more effective treatments are found. Meanwhile, electroencephalography remains the most useful investigation for diagnosis and prognosis. PMID- 12644747 TI - Epilepsy and behaviour. AB - PURPOSE OF REVIEW: Whilst behaviour disorder is so commonly a challenge to physicians in the field of epilepsy, comparatively little is known of its exact association with the epilepsies. This review highlights advances in knowledge, focussing on a broad definition of behaviour to include psychiatric pathology. RECENT FINDINGS: Differential rates of disturbance appear to be associated with concurrent intellectual disability. Advances in imaging, in particular, suggest that structural brain changes may underlie the increased susceptibility in some epilepsy patients. SUMMARY: Clinical practice should recognize that, with the possible exception of seizure freedom in postictal behavioural disturbance, epilepsy treatment alone is unlikely to treat the associated psychological and behavioural disturbance. Research continues to focus on assessing the correlates of epilepsy and psychopathology whilst the association between epilepsy and behaviour disorder in people with an intellectual disability remains largely unknown. PMID- 12644748 TI - Do epileptic seizures damage the brain? AB - PURPOSE OF REVIEW: The possibility that recurrent seizures in patients with poorly controlled epilepsy may produce neuronal damage and contribute to progressive functional and cognitive declines observed in some patients with epilepsy has major clinical and therapeutic implications for urgency of treatment and effective intervention to achieve complete control. RECENT FINDINGS: Advances in magnetic resonance imaging techniques, technical and conceptual advances in experimental analysis of neuronal death at the cellular and molecular level, and long-term neuropsychological observations have provided substantial new data and insights into phenomena of seizure-induced plasticity in neural circuitry that address the question "Do epileptic seizures damage the brain?" SUMMARY: The emerging perspective is that seizure-induced damage should be regarded not only as neuronal loss but as adverse long-term behavioral and cognitive consequences. This perspective provides a strong rationale for development of neuroprotective treatments to forestall adverse long-term consequences of repeated seizures, and for the importance of prompt, effective intervention that achieves complete seizure control. PMID- 12644749 TI - Mechanisms of antiepileptic drug resistance. AB - PURPOSE OF REVIEW: The present review considers new developments in the study and our understanding of resistance to treatment with antiepileptic drugs in epilepsy. RECENT FINDINGS: Studies suggest that mechanisms of resistance to drug treatment that operate in other diseases may also be relevant in human and animal drug-resistant epilepsy. Immunohistochemical and molecular genetic data show that there is overexpression of a number of genes and proteins that mediate nonspecific resistance to drug treatment. In particular, there is upregulation of P-glycoprotein and multidrug resistance-associated proteins 1 and 2. These proteins appear to be expressed in cerebral parenchymal cell populations that do not normally do so. Work in animal models suggests that these proteins are able to reduce the local concentration of antiepileptic drugs. Although these proteins are therefore good candidates for mediators of drug resistance, there is still limited proof that they are functionally important in human drug-resistant epilepsy. SUMMARY: Careful attention needs to be given to the identification and confirmation of the practical importance of mechanisms postulated to underlie drug resistance in human epilepsy. If certain mechanisms are shown to be involved, then new therapeutic options for drug-resistant human epilepsy may be forthcoming. PMID- 12644750 TI - New CNS-active drugs which are second-generation valproic acid: can they lead to the development of a magic bullet? AB - PURPOSE OF REVIEW: Valproic acid (VPA) is one of the four first line antiepileptic drugs (AEDs). VPA is also an effective drug in migraine prophylaxis and in treatment of bipolar disorders. The use of VPA is limited by its two rare but potentially life-threatening side effects, teratogenicity and hepatotoxicity, and it is the least potent of the established AEDs. Consequently, there is an incentive to develop a second-generation VPA. A successful, second-generation VPA would need to possess the following characteristics: broad-spectrum antiepileptic activity; better potency than VPA; and lack of teratogenicity and hepatotoxicity. These characteristics would give such a drug the potential to be utilized in epilepsy and other CNS disorders. RECENT FINDINGS: Intensive research has been carried out in order to develop a second-generation VPA that would be more potent and safer than VPA. Amide derivatives of VPA have shown particular value as potential follow-up compounds and have better in-vivo performance than VPA. Several CNS-active valproylamides are more potent as antiepileptics than VPA, they possess broad-spectrum antiepileptic activity, and have been found to be nonteratogenic in animal models. The amide analogues of VPA that emerged from structure-pharmacokinetic-pharmacodynamic relationship studies as promising second-generation compounds are: N-methyl-tetramethylcyclopropane carboxamide, (2S,3S)-valnoctamide, (R)-propylisopropyl acetamide and valproyl glycinamide. SUMMARY: At present there are three compounds in clinical trials in patients with epilepsy that can be regarded as second-generation VPA: valproyl glycinamide, 3 methylbutanamide or isovaleramide and SPD421 (DP-VPA). For any one of these second-generation valproic acids to become a successful follow-up compound to VPA, it has to fulfil the above criteria and also possess favorable pharmacokinetics. PMID- 12644751 TI - Emerging surgical and radiotherapeutic techniques for treating epilepsy. AB - PURPOSE OF REVIEW: Recent advances in epilepsy surgery have developed a resurgence of interest in the use of surgical techniques for the treatment of intractable epilepsy. RECENT FINDINGS: More invasive procedures such as hemispherectomy and multiple subpial transection have become more popular. Disconnective techniques such as multiple subpial transection have provided a surgical option for patients whose epileptogenic zone resides in the eloquent cortex. Alternatively, new minimally invasive neurostimulation therapies have been introduced to preserve maximal cerebral tissue. Radiosurgery has been recently utilized in the treatment of epilepsy with preliminary promising results. SUMMARY: In this analysis, the authors will attempt to review the more recent surgical approaches and their indications for the treatment of medically intractable epilepsy. For patients with the epileptogenic zone in the noneloquent cortex, seizure focus resection remains the most reasonable approach to therapy. PMID- 12644752 TI - Plaque radiotherapy for selected orbital malignancies: preliminary observations: the 2002 Montgomery Lecture, part 2. AB - PURPOSE: To describe the principles and preliminary results of plaque brachytherapy for selected orbital malignancies. METHODS: A custom-designed Iodine-125 plaque, designed to deliver a target dose of 50 Gray, was placed surgically in the region from which an orbital malignancy was partially resected. The mean dose to the target area was 50 Gray. The initial and follow-up patient data were reviewed. RESULTS: Of the 8 patients, the diagnosis was adenoid cystic carcinoma of the lacrimal gland (4 cases), orbital invasion by basal cell carcinoma (2), orbital extension of conjunctival melanoma (1), and metastatic carcinoma (1). Of the 4 with adenoid cystic carcinoma, there was microscopic residual tumor after excision and orbital exenteration was considered. Three have tumor control with follow-up of 1, 3, and 6 years. One patient required exenteration for recurrence separate from the field of brachytherapy and is free of tumor after 10 years. All 4 patients are alive and well with tumor control. Of the 2 patients with orbital extension of basal cell carcinoma, tumor control without recurrence has been achieved in both after 2 years. The patient with orbital metastasis responded to plaque radiotherapy, with no orbital recurrence, but died of systemic metastasis. The patient with orbital melanoma had local orbital recurrence separate from the area of irradiation and is currently being treated for systemic metastasis. CONCLUSIONS: Based on preliminary observations, plaque radiotherapy appears to be a reasonable alternative to exenteration and external irradiation for selected orbital malignancies. PMID- 12644753 TI - Margins of excision for cutaneous melanoma of the eyelid skin: the Collaborative Eyelid Skin Melanoma Group Report. AB - PURPOSE: To evaluate the practice patterns among surgeons who treat melanomas of the eyelid skin with respect to margins of excision and to look for possible correlation between margins of excision and the incidence of local and regional recurrence and distant metastasis. METHODS: A retrospective survey of the members of the American Society of Ophthalmic Plastic and Reconstructive Surgery and the European Society of Ophthalmic Plastic and Reconstructive Surgery yielded 44 cases. The patients' age, sex, date of diagnosis, histologic classification of melanoma, Breslow thickness, Clark level, location of melanoma, size of margins of excision, and findings of local or regional recurrence or distant metastasis were recorded in each case. Patients were stratified on the basis of margins of excision: 5 mm but <10 mm; and >/=10 mm. Patients were also stratified by Breslow thickness. A Cox regression model was used to evaluate the predictive value of each factor for recurrence. Main outcome measures were the incidences of local and regional recurrence and distant metastasis as a function of margins of excision and Breslow thickness. RESULTS: The majority of patients for whom reliable information was available had excision margins of /=10 mm, but this difference was not statistically significant because very few patients had melanomas at least 2 mm thick. Breslow thickness was the only statistically significant predictor of local, regional, and distant metastasis. Margins of excision did not have a statistically significant effect on local, regional, or distant recurrence. CONCLUSIONS: Breslow thickness is an important prognostic indicator for eyelid skin melanomas. A 5-mm margin of excision may be adequate for thin melanomas of the periocular skin, but because of the small number of patients in this series who had >5-mm margins, a definitive comparison of outcome with larger margins of excision cannot be made. For melanomas >/=2 mm, wider margins of excision may be prudent, and careful surveillance for local and regional recurrence is indicated. PMID- 12644755 TI - The effects of airbags on orbital fracture patterns in frontal automobile crashes. AB - PURPOSE: To investigate orbital fractures that occurred in frontal automobile crashes and to determine the effects of frontal airbags on injury incidence and severity. METHODS: The National Automotive Sampling System database files from 1993 to 2000 were examined. Frontal crashes were selected that included drivers and front-seat passengers only and excluded ejected occupants and rollover crashes. Orbital fractures could be closed, open, displaced, or any combination of these and were identified by using the Abbreviated Injury Scale codes. RESULTS: The analysis included 12,429,580 front-seat occupants from 25,464 cases. Of all occupants who were exposed to an airbag deployment, 0.09% sustained an orbital fracture. In contrast, occupants who were not exposed to an airbag deployment were more than twice as likely to sustain an orbital fracture (0.22%). In addition to reduction in incidence, airbags were also shown to decrease the severity of orbital fractures that occupants sustained. Occupants exposed to airbag deployment mostly sustained closed, less severe fractures (61.9%), whereas occupants not exposed to airbag deployment sustained the majority as more severe, open, displaced, or comminuted fractures (61.3%). CONCLUSIONS: This article presents the most comprehensive study of orbital fractures in automobile crashes to date. It is shown that both the incidence and the overall severity of orbital fractures decreases considerably with exposure to airbag deployment. This is accomplished because the airbag minimizes occupant contact with the windshield and steering wheel, which are the two leading sources of orbital fractures for occupants not exposed to airbag deployment. PMID- 12644754 TI - Feasibility of preoperative lymphoscintigraphy for identification of sentinel lymph nodes in patients with conjunctival and periocular skin malignancies. AB - PURPOSE: To determine the feasibility of preoperative lymphoscintigraphy for identification of sentinel lymph nodes (SLNs) in patients with conjunctival and periocular skin tumors and to determine the patterns of lymphatic drainage from such tumors. METHODS: We retrospectively reviewed the records of all patients with biopsy-confirmed conjunctival and periocular skin malignancies who underwent lymphoscintigraphy with or without SLN biopsy between January 1999 and June 2000. Patients underwent lymphoscintigraphy with 0.3 to 1 mCi of technetium Tc-99m sulfur colloid in a volume of either 0.2 mL or 1 mL. Images were taken as soon as the first SLNs were detected through the camera and every 15 minutes thereafter. Intraoperative mapping and SLN biopsy was performed 1 to 2 days after lymphoscintigraphy unless the patient refused or there were medical contraindications to the procedure. RESULTS: The study included 7 patients with malignant melanoma of the conjunctiva or periocular skin and 1 patient with Merkel cell carcinoma of the eyelid. On lymphoscintigraphy, at least 1 SLN was identified in 7 of the 8 patients. Although all lesions located in the lateral half of the ocular adnexa drained to at least one SLN in the parotid (preauricular) area, there was some variability in the drainage patterns of lesions located in the medial half of the ocular adnexa. A smaller injection volume (0.2 mL) was adequate for detecting the nodes draining the area of injection and led to less spread of technetium to the surrounding areas. Six patients underwent SLN biopsy. In all but one, the nodes identified during surgery corresponded with those visualized on lymphoscintigraphy. CONCLUSIONS: Preoperative lymphoscintigraphy successfully identifies SLNs in most patients with conjunctival and periocular skin malignancies. Smaller injection volumes (0.2 mL) appear to be adequate for identification of the sentinel nodes and lead to less spread to surrounding tissues. PMID- 12644756 TI - Balanced orbital decompression combined with fat removal in Graves ophthalmopathy: do we really need to remove the third wall? AB - PURPOSE: To compare the reduction of proptosis and the incidence of new-onset diplopia after 3-wall (medial, lateral, and inferior) orbital decompression versus balanced medial and lateral wall decompression combined with orbital fat excision in patients with Graves ophthalmopathy. METHODS: Three-wall orbital decompression including medial, inferior, and lateral walls was performed in 13 eyes of 7 patients (group 1), and balanced medial and lateral wall decompression combined with fat removal was performed in 18 eyes of 11 patients (group 2). A transnasal endoscopic approach was used for medial wall removal. A lateral canthotomy incision combined with a short upper eyelid incision was used for extended lateral wall removal, and this was combined with an inferior conjunctival fornix incision when floor decompression was performed. RESULTS: The mean reduction of proptosis was 6.9+/-1.6 mm and 6.5+/-1.3 mm in the first and second groups, respectively; the difference was not statistically significant (P=0.37). After 3-wall decompression, 57.1% of the patients had permanent new onset diplopia (group 1), whereas none of the patients had permanent postoperative diplopia after balanced medial and lateral wall decompression combined with fat removal (group 2). The difference in permanent new-onset postoperative diplopia between two groups was statistically significant (P<0.001). CONCLUSIONS: Balanced medial and lateral wall decompression combined with orbital fat removal provides an effective reduction in proptosis and reduces the incidence of postoperative permanent diplopia when compared with 3-wall decompression. This technique may eliminate the need for orbital floor excision. PMID- 12644757 TI - Prosthetic motility in pegged versus unpegged integrated porous orbital implants. AB - PURPOSE: To objectively measure and compare prosthetic motility in pegged versus unpegged orbital implants and to determine subjective patient assessment of motility after the pegging procedure. METHODS: A prospective case series of 10 patients with integrated porous orbital implants, who had secondary motility peg placement procedure, were studied. Infrared oculography was used to quantitatively assess pegged and unpegged prosthetic eye motility in horizontal and vertical excursions. RESULTS: For horizontal excursions, prosthetic motility in unpegged implants retained an average of 49.6% of measured motility of the contralateral normal eye, which increased to 86.5% with peg placement (P<0.05). For vertical excursions, prosthetic motility in unpegged implants retained an average of 51.3% of measured motility of the contralateral normal eye, which increased to 54.3% with peg placement (P>0.3). Nine of 10 patients judged their motility as "significantly improved," and 1 patient gave a rating of "some improvement" after peg placement. Four of 10 patients had granulomas around the peg sites. CONCLUSIONS: Objective assessment of prosthetic motility shows a significant increase in horizontal gaze after motility peg placement. PMID- 12644758 TI - Improved prosthetic motility with placement of a second coupling PEG. AB - PURPOSE: Integrated coupled orbital implants have been shown to improve prosthetic motility over noncoupled orbital implants. Some patients are limited in prosthetic motility despite this coupling for various mechanical and physiological reasons. A common limitation of motility in anophthalmic rehabilitation is forniceal constriction or scarring. Anophthalmic patients with coupled integrated implants who were noted to have limited prosthetic motility because of conjunctival scarring, improperly placed coupling pegs, rotation of the prosthesis, or contracture in the fornices underwent placement of a second coupling peg with the aim of improving prosthetic motility. METHODS: Six patients were selected who fit the criteria of poor prosthetic motility despite excellent implant motility. All patients had previously placed hydroxyapatite implants followed by motility peg placement several months later. Titanium motility peg placement was only placed after gadolinium-enhanced magnetic resonance imaging neuroimaging studies were performed to assess proper vascularization of the integrated implant. After determining the direction of motility limitation caused by conjunctival dysfunction, a second titanium motility peg was placed in the standard fashion to improve prosthetic motility. RESULTS: All six patients had improvement of prosthesis motility, as noted by both the patient and surgeon, after second motility coupling peg placement. No complications were encountered. CONCLUSIONS: In select patients with poor translation of implant motility to prosthesis motility, placement of a second motility coupling peg may improve prosthetic motility. PMID- 12644759 TI - The bioceramic orbital implant: experience with 107 implants. AB - PURPOSE: To assess the problems associated with the Bioceramic (Aluminum oxide, Al(2)O(3)) orbital implant. METHODS: A consecutive case series of 107 patients receiving a Bioceramic orbital implant by two surgeons over a 3-year period were reviewed. The authors analyzed patient age, type of surgery, size of implant, peg system, follow-up duration, time of pegging, complications encountered, and treatment. RESULTS: Seven patients were lost to follow-up after 2 months, leaving 100 patients who were followed from 3 to 38 months (average, 13.4 months). Three patients died during the follow-up period (one with 2 months' follow-up, one with 7 months' follow-up, and the third with 9 months' follow-up). Implant-related problems occurred in 11 (11%) patients. Discharge occurred in 5 (5%) patients, implant exposure in 2 (2%), socket discomfort in 1 (1%), trochleitis in 1 (1%), and conjunctival thinning in 2 (2%). Peg problems occurred in 11 (34.3%) of 32 pegged patients: discharge in 4 (12.5%) and in 1 patient (3.1%) each, pyogenic granuloma, conjunctiva overgrowing the peg, implant exposure around the sleeve, clicking, accumulation of black material, and broken peg during insertion. Infection did not occur in any patient. CONCLUSIONS: The Bioceramic orbital implant represents an alternative porous orbital implant that is biocompatible with orbital tissues, easy to manufacture, structurally strong, and less expensive than other commercially available porous orbital implants (e.g., Bio Eye hydroxyapatite implant). Problems encountered with its use are similar to those seen with the Bio-Eye orbital implants but appear to occur less often. The incidence of exposure associated with the Bioceramic implant is less than that reported for the Bio-Eye. PMID- 12644760 TI - Comparison of synthetic hydroxyapatite and porous polyethylene implants in eviscerated rabbit eyes. AB - PURPOSE: Synthetic hydroxyapatite and porous polyethylene (Polipore) spheres were placed in rabbits' eviscerated cavities to evaluate tissue reaction and volume maintenance. METHODS: Fifty-six Norfolk albino rabbits underwent unilateral evisceration and implantation of synthetic hydroxyapatite (H group, 28 animals) or porous polyethylene spheres (P group, 28 animals). Postoperative reactions, animal behavior, and socket conditions were monitored. Light microscopy and morphometric evaluation with statistical analysis of the exenterated orbits were performed at 7, 15, 30, 60, 90, 120, and 180 days. Scanning electron microscopy was appraised 7, 60, and 180 days after surgery. RESULTS: Two animals from the H group and 1 from the P group had extrusion 7 days after surgery. Throughout the experimental period, the synthetic hydroxyapatite caused more inflammation than the porous polyethylene material. Ingrowth in the sphere occurred 7 to 15 days after the surgery in both groups, and the tissue reaction became denser at approximately 60 to 90 days, when bony metaplasia began in the H group. Volume maintenance was better in the P group and with a smaller pseudocapsule surrounding the implanted sphere than in the H group. CONCLUSIONS: Clinical findings demonstrated mild inflammation inside the sphere and in the pseudocapsule surrounding it and better cavity volume maintenance in the P group animals. The authors consider porous polyethylene a more suitable material than synthetic hydroxyapatite for use in anophthalmic cavity reconstruction. PMID- 12644761 TI - Treatment of recurrent eosinophilic granuloma with systemic therapy. AB - PURPOSE: To describe the systemic treatment of recurrent eosinophilic granuloma. METHODS: Case report and review of Langerhans cell histiocytosis. RESULTS: A 9 year-old boy had painful, right upper eyelid swelling. A computed tomography scan showed a right superolateral orbital mass with evidence of bony erosion. Frozen section evaluation was consistent with eosinophilic granuloma. Biopsy and curettage were performed. Systemic work-up was negative for extraorbital disease. Recurrence of the orbital mass 6 weeks after surgery was confirmed by magnetic resonance imaging (MRI). Four weeks after treatment with prednisone and vinblastine, MRI showed a marked decrease in the size of the orbital lesion. Additional treatment with 6-mercaptopurine and methotrexate was initiated, and MRI 6 months later showed complete resolution of the lesion. CONCLUSIONS: The treatment of recurrent eosinophilic granuloma is challenging. The goal is to effectively cure the lesion with minimal short-term and long-term side effects. Systemic steroids, vinca alkaloids, antimetabolites, and antifols are preferred because they have less toxicity and mutagenicity. Combination chemotherapy used in conjunction with treatment by a pediatric hematologist/oncologist appears to be an effective method of treating recurrent disease. PMID- 12644762 TI - Perspective: the physical basis of surgical electrodissection. PMID- 12644763 TI - Topical mitomycin treatment for primary acquired melanosis of the conjunctiva. AB - A case of primary acquired melanosis of the conjunctiva primarily treated with topical mitomycin chemotherapy is presented. The patient underwent treatment with one 2-week course of topical mitomycin 0.02% four times daily, followed 4 weeks later by a 2-week course of topical mitomycin 0.04% four times daily, followed by a 3-month course of topical mitomycin 0.02% twice daily. Map biopsies were performed after the initial two courses of mitomycin therapy and 12 months after all courses of mitomycin treatment had been completed. Complete resolution of the conjunctival pigmentation was noted after the final course of treatment. Map biopsies performed at 12 months following completion of treatment showed normal conjunctiva histopathologically. No recurrence of pigmentation was noted in follow-up 26 months after completion of treatment. Topical mitomycin chemotherapy may be a successful primary treatment for primary acquired melanosis of the conjunctiva. PMID- 12644764 TI - Dermatochalasis causing apparent bitemporal hemianopsia. AB - We describe a previously unreported cause of apparent bitemporal hemianopsia. Three patients with bilateral dermatochalasis underwent Humphrey visual field (HVF) testing in anticipation of corrective blepharoplasty. Unexpectedly, the HVF showed bitemporal hemianopsia in each case, prompting magnetic resonance imaging of the brain in one case. The scan was negative. The HVF tests were repeated with the upper eyelids taped in elevation, and were normal. We conclude that the bitemporal hemianopic defects in our patients resulted from the testing strategy employed by the HVF analyzer software to measure the visual field. Reasons for these artifactual results are discussed. Clinicians should be aware of this uncommon cause of bitemporal hemianopic visual field defects. PMID- 12644765 TI - Proptosis after hair pulling. AB - We describe proptosis due to a subperiosteal orbital hematoma that originated from a subgaleal bleed caused by hair pulling. A 13-year-old boy presented with a one-week history of progressive proptosis after his older sister pulled his hair during an argument. Computerized tomography showed a subgaleal hematoma and a right superior subperiosteal orbital hematoma. Progressive proptosis and compressive optic neuropathy developed, necessitating surgical evacuation with a favorable outcome.Hair pulling can result in a subgaleal bleed with extension to the orbital subperiosteal space. When managing patients with a subgaleal hematoma, this vision-threatening complication should be considered and treated accordingly. PMID- 12644766 TI - Bilateral dynamic proptosis due to frontoethmoidal sinus mucocele. AB - A 34-year-old man presented with bilateral inferior and lateral globe displacement and spontaneous, dynamic proptosis in both eyes, pulsatile in nature. Computed tomography and magnetic resonance imaging scans confirmed bilateral frontoethmoidal mucoceles with erosion of the orbital roof, and intracranial extension. Following a transcutaneous mini-trephine of the left orbital mucocele, the dynamic proptosis disappeared, and both proptosis and restriction of eye movements in the left eye improved. This is the first reported case of frontoethmoidal sinus mucocele causing bilateral spontaneous dynamic proptosis. PMID- 12644767 TI - Exophthalmos and orbital floor thickening related to maxillary sinusitis. AB - A 59-year-old woman presented with periocular pain and 1.5 mm of relative proptosis. A computed tomography scan demonstrated thickening of the orbital floor, and an endoscopic sinusectomy revealed chronic sinusitis. Although cases of maxillary sinus disease with orbital floor thickening have been reported, these findings are in contrast to cases of silent sinus syndrome, in which maxillary sinus disease induces thinning and inferior displacement of the orbital floor with secondary enophthalmos and hypoglobus. Thus, while chronic maxillary sinus disease may create negative antral pressure and inferior displacement of the orbital floor, whether orbital floor thinning or thickening occurs varies with each case. The present case represents a combination of maxillary sinusitis with orbital floor thickening and exophthalmos, a less common manifestation of maxillary sinus-related orbitopathy. PMID- 12644769 TI - Recurrent solitary reticulohistiocytoma of the eyelid. AB - A healthy 13-year-old female presented with a 5-month history of an enlarging red brown, firm lower eyelid mass associated with loss of eyelashes. An excisional biopsy revealed a well-circumscribed nodule composed of coalescing nests of epithelioid cells, rare multinucleated cells, and vacuolated macrophages and spindle cells. Additional studies revealed a positive vimentin and alpha(1) antitrypsin, and negative S100 protein and Factor XIIIa immunophenotype. This constellation of clinicopathologic features is diagnostic of solitary reticulohistiocytoma, but also shows overlap with xanthogranuloma, a common childhood histiocytosis. Reticulohistiocytoma should be considered in the differential diagnosis of eyelid tumors and has the potential for local recurrence. Diffuse cutaneous reticulohistiocytosis or multicentric reticulohistiocytosis should be considered if similar cutaneous lesions with or without symptoms of arthritis are present, respectively. PMID- 12644768 TI - Primary diffuse large B-cell lymphoma of the lower eyelid. AB - An 80-year-old woman presented with a palpable mass in the central right lower eyelid, with no visible alterations of the overlying skin or eyelid margin, and no signs of inflammation. Eversion of the eyelid revealed an ulcerating lesion approximately 8 mm in diameter, suggesting malignant lesion. Excision with eyelid reconstruction using the Hughes technique was performed. Histopathological analysis revealed a diffuse infiltrate with large atypical lymphocytes. Borders of the surgical resection were free from infiltration. Histochemical staining of the proliferating cells with monoclonal antibodies was positive for CD20 and CD79a, and polyclonal antibodies were positive for Lambda chains, confirming a diffuse large B-cell lymphoma. After 24 months' follow-up there has been no local or systemic recurrence of the disease. Lymphoma may present as an ulcerating lesion of the tarsal conjunctiva. Complete surgical excision, with no evidence of systemic disease, may be considered to carry a good prognosis and survival. PMID- 12644770 TI - Recurrent facial palsy occurring with metastatic thymic carcinoid and nonarteritic ischemic optic neuropathy. AB - A 74-year-old man was diagnosed with thymic carcinoid metastatic to the mediastinum. Two years later, he developed left and then right facial palsy. Four additional episodes of facial palsy developed within 2 years, followed by an occurrence of nonarteritic ischemic optic neuropathy. While recurrent facial palsy is uncommon, it may occur in the presence of various systemic conditions such as carcinoid, diabetes, infection, and neoplasm. The presence of recurrent facial palsy logically warrants an evaluation for systemic disease. PMID- 12644771 TI - Transitional cell carcinoma of the urinary bladder metastatic to the orbit. AB - We report a rare case of transitional cell carcinoma of the urinary bladder metastatic to the orbit, review the histopathology, and highlight characteristics of the few previously reported cases. A 68-year-old man with a remote history of transitional cell carcinoma of the urinary bladder presented with decreased visual acuity, afferent pupillary defect, and limited extraocular motility OS. Computed tomography scan of the orbits showed a diffuse infiltrate surrounding the left globe and optic nerve. Biopsy revealed metastatic transitional cell carcinoma. The patient expired one month later. With only seven cases previously reported, we conclude that transitional cell carcinoma of the urinary bladder rarely metastasizes to the orbit. When present, orbital metastases signify aggressive disease and a poor prognosis. Mean survival following orbital diagnosis is 2.2 months in the cases reported. PMID- 12644772 TI - Fostering applications of genetics in primary care: what will it take? PMID- 12644773 TI - Family history: where to go from here. PMID- 12644774 TI - Diffusion of innovations. PMID- 12644775 TI - Barriers to the provision of genetic services by primary care physicians: a systematic review of the literature. AB - PURPOSE: The purpose of this literature review is to report primary care physicians' perceived barriers concerning the provision of genetic services. METHODS: Sixty-eight papers were identified in six electronic databases. Only publications classified as empirical studies (N = 18) were included in this review. RESULTS: Barriers identified most frequently in reviewed studies were inadequate knowledge of basic genetics, lack of detailed or updated family histories, lack of confidence, and lack of referral guidelines. CONCLUSION: Although many primary care physicians perceive genetics as a low practice priority, they do report a need for educational programs in genetics, informational resources, and referral guidelines. PMID- 12644776 TI - Prospective assessment in newborns of diabetes autoimmunity (PANDA): maternal understanding of infant diabetes risk. AB - PURPOSE: To assess accuracy of mothers' understanding of their newborns' genetic risk for type 1 diabetes and to identify predictors of the comprehension and retention of genetic information. METHODS: Mothers of 435 newborns genetically screened at birth were informed of the infant's risk for type 1 diabetes using a standard script that provided both categorical and numerical risk information. The mothers' comprehension and retention of this information were assessed by structured interview on two occasions, approximately 3.6 weeks and approximately 3.9 months postnotification. RESULTS: At the initial interview, 73.1% of mothers gave a correct estimate of their child's genetic risk, 3.2% overestimated risk, 13.3% underestimated risk, and 10.3% could not recall risk at all. At the follow up interview, fewer mothers (61.9%) correctly estimated their child's risk and more mothers (24.4%) underestimated their child's risk. Maternal accuracy was associated with maternal education, ethnic minority status, infant risk status, maternal ability to spontaneously recall both categorical and numerical risk estimates, and length of time since risk notification. Underestimation of risk was associated with maternal education, family history of diabetes, time since risk notification, and maternal anxiety about the baby's risk. CONCLUSION: The accuracy of mothers' recall of infant risk declines over time, with an increasing number of mothers underestimating the infant's risk. Effective risk communication strategies need to be developed and incorporated into genetic screening programs. PMID- 12644777 TI - The genetic family history as a risk assessment tool in internal medicine. AB - PURPOSE: The study goals were to (1) determine the proportion of unselected individuals at increased risk for diseases with known genetic components and (2) compare the documentation and quality of risk assessment between a questionnaire, a pedigree interview, and chart review. METHODS: Seventy-eight patients seen in a division of internal medicine were randomized into two groups, which completed a questionnaire or underwent a pedigree interview. Chart notes were compared to both study tools. RESULTS: Sixty-two (79.5%) of the 78 participants scored at increased risk for at least one category. Either of the two study tools found significantly more people at high risk (48/78, 61.5%) than the chart review (31/78, 39.7%) (P = 0.01). CONCLUSIONS: Approximately 20% of patients in an unselected internal medicine practice were at an increased risk that was not documented in reviewed chart notes. Targeted family history analysis reveals patients who require increased medical surveillance, preventive measures, or genetic counseling/testing. PMID- 12644778 TI - Early adoption of BRCA1/2 testing: who and why. AB - PURPOSE: Relatively little is known about the characteristics of women who chose to undergo BRCA1/2 testing soon after testing became available, including how they became aware of and chose to pursue testing. Diffusion of innovation theory states that acceptance of an innovation is a function of the potential adopter's value for innovation and perceptions of the compatibility, complexity, and relative advantage of the innovation. These factors may contribute to early uptake of BRCA1/2 testing. The purpose of this study was to describe the characteristics of women who were "early adopters" of clinical BRCA1/2 testing and to determine whether diffusion theory explains variation in uptake of testing after participation in genetic counseling for BRCA1/2 testing. METHODS: A retrospective cohort study of participants in genetic counseling for BRCA1/2 testing was conducted at a clinical program in a large academic health system. Measures included components of diffusion of innovation theory (participant's value for innovation, i.e., "innovativeness," and perceptions of the compatibility, complexity, and relative advantage of BRCA1/2 testing), characteristics of how the participant became aware of and sought BRCA1/2 testing, and decisions about testing after counseling. RESULTS: From the 229 respondents, 71 (31%) had undergone testing at the time of the survey. Fifty seven women (25%) had sought BRCA1/2 testing because a family member had breast or ovarian cancer and 37 (16%) because they had breast or ovarian cancer. Only 15 women (7%) reported seeking testing because of a physician's recommendation. After multivariate adjustment, higher innovativeness and higher ratings of the compatibility of BRCA1/2 testing were associated with undergoing testing after counseling [relative risk (RR) 1.76, 95% confidence interval (CI) 1.2-2.6]. However, ratings of the complexity or relative advantage of testing were not associated with testing decisions. Higher innovativeness was associated with being the first in the family to undergo testing (RR 4.85, 95% CI 1.6-14.9), becoming aware of BRCA1/2 testing through the media (RR 1.50, 95% CI 1.0-2.4), and being aware of BRCA1/2 testing prior to counseling (RR 1.25, 95% CI 1.1-1.4). CONCLUSIONS: The uptake of BRCA1/2 testing among women undergoing genetic counseling was associated with innovative characteristics of the participant and the perceived compatibility of the test with existing values and needs, but not with the complexity or relative advantage of the test. Most "early adopters" had heard of BRCA1/2 testing from a source other than their physician and had sought testing because of a personal or family member's cancer diagnosis. These findings can inform predictions surrounding the introduction of future genetic susceptibility tests and strategies for guiding the further diffusion of BRCA1/2 testing. PMID- 12644779 TI - Does knowledge about the genetics of breast cancer differ between nongeneticist physicians who do or do not discuss or order BRCA testing? AB - PURPOSE: To assess nongeneticist physicians' knowledge and experience with BRCA1/2 testing. METHODS: In 1998, 2250 internists, obstetrician-gynecologists (Ob-Gyns), and oncologists practicing in Pennsylvania, Maryland, Massachusetts, New York, or New Jersey were surveyed. RESULTS: Forty percent responded. Only 13% of internists, 21% of Ob-Gyns, and 40% of oncologists correctly answered all four knowledge questions about genetic aspects of breast cancer and testing for it. Knowledge was associated with discussing or ordering only among oncologists. CONCLUSION: Despite deficiencies in their knowledge about the genetic aspects of breast cancer, many nongeneticist physicians have discussed testing and some have ordered testing. PMID- 12644781 TI - Thrombocytopenia in patients with 22q11.2 deletion syndrome and its association with glycoprotein Ib-beta. AB - PURPOSE: To elucidate whether thrombocytopenia in 22q11.2 deletion syndrome patients is associated with the hemizygosity of glycoprotein Ib-beta and to clarify the correlation of phenotype and genotype of this gene in 22q11.2 deletion syndrome patients with thrombocytopenia. METHODS: Platelet number, mean platelet volume, platelet agglutination, and the protein level of glycoprotein Ib beta were measured in 22q11.2 deletion syndrome patients and controls. Phenotypes other than that of thrombocytopenia were also analyzed in these patients. RESULTS: The 22q11.2 deletion syndrome patients with thrombocytopenia had a larger mean platelet volume, lower agglutination to ristocetin, and lower protein level of glycoprotein Ib-beta than control patients. The 22q11.2 deletion syndrome patients with thrombocytopenia showed an increased risk of developing schizophrenia. CONCLUSIONS: Thrombocytopenia in 22q11.2 deletion syndrome patients is associated with decreased expression of glycoprotein Ib-beta because of the hemizygosity. 22q11.2 deletion syndrome patients with thrombocytopenia require total management, especially for schizophrenia. PMID- 12644780 TI - Attitudes of the broader hearing, deaf, and hard-of-hearing community toward genetic testing for deafness. AB - PURPOSE: To assess attitudes in a nonmedically and nonculturally influenced setting of reproductive-age adults toward genetic testing for deafness in newborns. METHODS: Hearing, deaf, and hard-of-hearing individuals at a university completed questionnaires assessing attitudes toward genetic testing. RESULTS: Eighty-five percent of hearing (n = 133) and 62% of deaf/hard-of-hearing (n = 89) individuals would allow genetic testing for deafness in their own newborn. CONCLUSIONS: These results indicate an acceptance of newborn genetic testing for deafness by individuals in the broader community, regardless of hearing status. PMID- 12644782 TI - Hereditary, familial, and idiopathic chronic pancreatitis are not associated with polymorphisms in the tumor necrosis factor alpha (TNF-alpha) promoter region or the TNF receptor 1 (TNFR1) gene. AB - PURPOSE: Genetic alterations that are associated with acute and chronic pancreatitis remain to be identified. METHODS: The authors investigated two functionally active tumor necrosis factor (TNF) promoter region polymorphisms at positions -238 and -308 and the entire coding region of the corresponding TNF receptor 1 (TNFR1) gene in 54 patients with hereditary, familial, and idiopathic chronic pancreatitis who were previously tested negative for cationic trypsinogen mutations by direct DNA sequencing. RESULTS: In three patients, we detected novel DNA variants in the TNFR1 gene that did not segregate with the disease. The genotype frequencies of the TNF promoter polymorphisms were similar between patients and controls. CONCLUSION: These polymorphisms are not associated with hereditary, familial, or idiopathic chronic pancreatitis. PMID- 12644784 TI - [Committee on cancer of the upper aerodigestive tract and survey on buccodental aspects. Report of 164 teams]. AB - BACKGROUND: Head and neck cancer occupies an important place in the field of oncology in France (8%). Patient management requires a wide diversity of treatment modalities and must be based on a multidisciplinary approach. MATERIALS AND METHODS: This clinical ethics study was based on analysis of a national survey of current practise in public and private hospital structure. A 28 questions anonymous questionnaire was send to 164 practitioners caring for these patients from the stage of diagnosis and evaluated the modalities of dental treatments and the related ethical aspects. RESULTS: The competence of the various practitioners in their respective fields is undeniable, but there is a lack of multidisciplinary collaboration. The absence of an oral practitioner is a serious problem and can reduce the patient's chances of cure. DISCUSSION: The oral practitioner must participate in head and neck cancer committees at all steps of treatment in order to provide curative, preventive care and prosthetic rehabilitation, thereby contributing to improvement of the quality of life of these patients. PMID- 12644785 TI - [Squamous-cell carcinoma of the tongue: treatment results and prognosis]. AB - OBJECTIVE: The aim of the study was to assess the results of curative treatment of patients with squamous cell carcinoma of the tongue and to evaluate survival and predictive factors of recurrence. PATIENTS AND METHODS: A series of 309 patients with squamous cell carcinoma of the tongue treated with curative intent was studied from January 1988 to December 1999. The percentage of oral tongue cancer was 82.2 and the percentage of cancer of base of the tongue was 17.8. Most patients underwent surgical procedure alone or combined with radiotherapy (92%). We performed 252 neck dissections. Bilateral dissections were performed for cancer of the apex linguae, cancer of the base of the tongue, for patients with N2c neck disease and whenever the primary tumor site crossed the median line. Twenty-five patients (8%) were treated with radiation therapy alone. Mean follow up was 55 months. The functional results were assessed within a minimum of 10 months postoperative follow-up. RESULTS: In 45.2%, there was histological evidence of node invasion with 53.5% of extracapsular node spread in the neck specimens. Extracapsular node spread did not influence survival or recurrences. Occult cervical metastasis in an elective neck dissection in clinically negative necks was found in about 20% of patients with 47% of extracapsular node spread (41% for cancer of mobile tongue and 80% for those of base of the tongue). About 23% of patients with cancer of base of the tongue staged N0 had histological node invasion in controlateral neck nodes. The postoperative mortality rate was 0.9%. The rate of complications was 17%. The cancer recurred in 41.7% of all cases. Twelve percent of all patients had second primary cancers of the upper aerodigestive tract. The overall survival and non-recurrence rates at 2 and 5 years were higher in cancer of oral tongue than in cancer of base of the tongue. Survival rates were better when neck nodes were clinically or histologically negatives and in early-stage carcinomas. Non-recurrence rates were better when nodes were clinically or histologically negatives and when margins of exeresis were not involved. The functional results were better in oral tongue cancer than in base of the tongue cancer. DISCUSSION: Prognosis (survival and non-recurrence rates and functional results) of squamous cell carcinomas of oral tongue was better than prognosis of those of base of tongue. We recommend an aggressive surgical procedure even in patients with neck classed N0 (with reservations for T1 lesions with small depth of invasion): an ipsilateral supraomohyoid neck dissection for cancer of oral tongue and a bilateral supraomohyoid neck dissection for cancer of base of the tongue, cancer of oral tongue which crosses the median line of the oral cavity and cancer of the apex linguae. Postoperative radiotherapy must be performed when margins are positives and/or when nodes are involved with or without extracapsular spread. PMID- 12644786 TI - [Reconstruction of the mobile tongue malignant tumor excision: quality-of-life assessment in 19 patients]. AB - INTRODUCTION: Tumor tongue excision leads to anatomical and functional defects (chewing, swallowing, oral and prosthetic hygiene). The extension of the resection depends on tumor size and surgical radicality. MATERIAL AND METHOD: In our department, 72 cases of lingual tumor were treated between may 1987 and January 1997. 73% of these cases were infiltrating squamous cell carcinomas. 71% were male. Most of the tumors were located at the ventral (45%) and lateral side (36%). 25% of the tumors crossed the midline. Staging was most often pT(2) (46%) and pN(0) (62%). Reconstruction with radial free flap occurred in 37%, with pedicled pectoralis major muscle flap in 13% and with pedicled latissimus dorsi muscle flap in 25% of cases. 51% of the patients were irradiated. All the patients were asked about their quality of live by an 11 questions questionnaire. The questions concerned the different oral and lingual functions and the social, familial, affective and professional life. RESULTS: 19 patients (that means 26%) answered the questionnaire. 9 of them were reconstructed with a flap (6 free radial flaps, 2 pedicled pectoralis major flaps, 1 pedicled latissimus dorsi flap) and were irradiated. In the 10 other patients wounds were closed by simple sutures of the margins. 1 of these 10 patients was irradiated. There was no correlation between scores of the questions and neither pT nor age of patients. We observed a parallelism between scores of the questions about speech and swallowing. Scores of the questions about swallowing were higher when a flap was used for reconstruction, whatever was the type of flap. DISCUSSION: A questionnaire was drawn up from the UW QOL questionnaire. It was a simplified questionnaire with reduced number of questions and simple words. Lesser tongue immobility leads to a better swallowing. Swallowing and speech were worst when tongue resection was large. In this quality of life study, there was a few number of patients that answered the questionnaire so that it is impossible to make a difference between the different flaps. PMID- 12644787 TI - [Retrospective analysis of 30 cases of mandibular ameloblastoma operated in the Ivory coast from 1992 to 2000]. AB - OBJECTIVE: We analyzed a retrospective series of 30 cases of ameloblastoma operated in the Ivory coast from 1992 to 2000. PATIENTS AND METHODS: The following techniques were used--temporary reconstructions via the endobuccal approach using a Peri prothesis in 27 cases and a resin coated Peri prosthesis in one;-- definitive reconstructions using iliac bone grafts in 3 cases and rib grafts in 14. These reconstructions were performed in a second operation via a cutaneous approach to replace the Peri prosthesis. Thirteen patients were lost to follow-up for this second phase. Mean follow-up ranged from 6 months to 13 months (mean 9.9 months). RESULTS: Satisfactory outcome was acchevied when mouth opening was greater than 10 mm and patients with an acceptable cosmetic result no longer complained of mastication or phonation problems. There were four postoperative infections after bone graft reconstruction (two of them were immediate reconstruction). The resin coated Peri prosthesis led to a skin ulceration. DISCUSSION: This study demonstrated the interest of mandibular reconstruction in undermedicalized countries where indications are dominated by reconstruction after resection for ameloblastoma. The large number of patients lost to follow-up before the second stage bone grafts points out the need for immediate reconstruction using autologous bone. Further study in a larger series would be needed to determine the cause of failure after immediate reconstruction. PMID- 12644788 TI - [Indications for dentofacial and dentomaxillofacial orthopedic care in children and adolescents--June 2002 ANAES]. PMID- 12644789 TI - [Paradontopathy: diagnosis and treatment--May 2002]. PMID- 12644790 TI - [Optimizing Lipostructures: applications in maxillofacial surgery]. PMID- 12644791 TI - [On post-traumatic ptosis]. PMID- 12644792 TI - [Emergency reimplantation of the tongue after complete traumatic amputation]. AB - Complete traumatic amputation of the tongue is very exceptional, the intra-oral position generally protecting the tongue. Most reported cases of trauma have involved partial section with satisfactory results after simple suture. Complete amputation of a portion of the tongue occurred in a 12-year-old boy during a fight. The fragment of the tongue was brought to the emergency unit. It was placed in heparinated saline solution and replanted successfully three hours after the trauma. Plastic surgery of the oral cavity was also performed to repair laceration injuries. The postoperative period was uneventful and the tongue regained vitality and normal motion. There was just a slight retraction. Replantation of the tongue after complete section may not be successful but should be attempted when possible. PMID- 12644794 TI - Abutment positioning in a cantilevered shortened dental arch: a clinical report and static analysis. AB - This article describes the extension of a shortened dental arch with a cantilever fixed partial denture in a patient who refused to be treated with an implant supported and retained prosthesis. Orthodontic movement of the distal abutment to enlarge the interabutment span was performed before the final restoration. The mechanical advantage of the enlarged span in a cantilever fixed partial denture is analyzed, discussed, and demonstrated by 2-dimensional static analysis. PMID- 12644795 TI - Fracture of the genial tubercles associated with a mandibular denture: a clinical report. AB - Fracture of the genial tubercles without an associated mandibular fracture is uncommon. It occurs mainly in patients wearing a complete mandibular denture when the mandible is atrophied and the genial tubercles are hypertrophied. Only 7 reports have been found in the English-language literature. An additional situation of a 70-year-old woman is presented along with a review of the literature. A conservative approach to treatment is advised with this type of fracture. The use of dental implants should be considered in patients with hypertrophied tubercles to prevent such fracture. PMID- 12644796 TI - Prosthetic rehabilitation of a total rhinectomy patient resulting from squamous cell carcinoma of the nasal septum: a clinical report. AB - The clinical characteristics and prosthetic rehabilitation of a patient with squamous cell carcinoma of the nasal septum after combined radiation therapy and a total rhinectomy is presented. PMID- 12644797 TI - An alternate treatment method for a fixed-detachable hybrid prosthesis: a clinical report. AB - The inability to consistently achieve a passive fit with screw-retained implant prostheses is well documented in the literature. Cement-retained implant frameworks have been advocated for implant-retained fixed partial dentures. However, cement retention has not been routinely advocated for fixed-detachable hybrid prostheses. This article describes a method for fabricating a cement retained fixed-detachable hybrid prosthesis. The advantages and disadvantages are discussed. PMID- 12644798 TI - The anterior cantilever in the implant-supported screw-retained mandibular prosthesis. AB - STATEMENT OF PROBLEM: When implants are placed interforaminally in mandibular implant-supported screw-retained prostheses, the most anterior implant is usually positioned lingual to the incisors of the prosthesis. This creates an anterior cantilever with the entire prosthesis acting as a class I lever and possibly placing the anterior implant under alternating tension and compression during function. PURPOSE: The purpose of this study was to measure the anterior cantilever of randomly chosen patients with restored mandibular implant-supported fixed prostheses, and to establish the proportions of anterior to posterior cantilever lengths relative to the anteroposterior spread. MATERIAL AND METHODS: Thirteen edentulous patients were recruited from the University of Minnesota's dental implant program. Each patient had been restored with a maxillary complete denture and a mandibular implant-supported screw-retained prosthesis supported by 5 endosseous implants. Each patient had 1 mandibular impression made with irreversible hydrocolloid, which was poured in type III gypsum. A FaroArm precision 3D measuring stylus was used to make multiple-axis (X-Y-Z) measurements (mm) on the casts of the dental implants, of anteroposterior spread, and anterior and posterior cantilevers. Presence or absence of screw loosening was noted using a screwdriver with finger pressure. Data were analyzed using a 1-way analysis of variance to compare prostheses with loose screws to prostheses without loose screws, for each of 3 outcome measures: length of anterior cantilever, length of posterior cantilever, and anteroposterior spread (P<.05). RESULTS: Mandibular anterior cantilever lengths ranged from 5.5 to 14.4 mm with a mean of 8.78 mm. Combined posterior cantilever lengths ranged from 9.2 to 20.9 mm with a mean of 16.2 mm. Anteroposterior spread ranged from 5.2 to 12.3 mm with a mean of 7.9 mm. From a total of 65 retaining screws, 7 were found to be completely loose. No apparent correlation was found between length of mandibular anterior cantilever and screw loosening (P=.45), although the ratio of posterior cantilever to anteroposterior spread (2:4) was significantly associated with screw loosening (P=.006). CONCLUSION: Within the limitations of this study, anterior cantilevers in mandibular implant-supported screw-retained prostheses were frequent and appeared to depend on implant placement and prosthesis design. The ratio of anterior to posterior cantilever lengths was approximately 1:2. PMID- 12644799 TI - Accuracy of open tray implant impressions: an in vitro comparison of stock versus custom trays. AB - STATEMENT OF PROBLEM: The accuracy of an implant fixture-level impression is affected by the type of impression tray used. PURPOSE: The purpose of this in vitro study was to investigate the accuracy of open tray implant impressions comparing polycarbonate stock impression trays and rigid custom-made impression trays to make implant fixture-level impressions. MATERIAL AND METHODS: Gold cylinder pairs, splinted by gold bars (reference frameworks) were constructed on an aluminum typodont. Polyether impressions were made of 2 pairs of Branemark 3.75-mm diameter fixtures mounted in an aluminium typodont, with 3 stock impression trays, 3 close-fit custom trays, and 3 spaced custom impression trays, by use of an open tray technique. The casts produced were assessed for accuracy by attaching the reference frameworks with alternate single screws and measuring the vertical fit discrepancy of these reference frameworks to the analogs within the working cast using a traveling microscope. Comparison of gap dimensions by tray type was performed with a nonparametric Kruskal-Wallis analysis of variance (ANOVA) followed by pair-wise Mann-Whitney U tests. To adjust for multiple comparisons in the post-ANOVA contrasts (P<.02). Comparison of gap dimensions between anterior and posterior regions were performed with Mann-Whitney U tests (P <.05). RESULTS: The results showed that the mean fit accuracy, as measured by vertical fit discrepancy, of casts from the stock trays (23 +/- 20 microm) were statistically significantly less (P<.001) than the spaced custom trays (12 +/- 10 microm) or close fit custom trays (11 +/- 10 microm). The difference in median gap size for analogs with a 20-mm separation was 10 microm. CONCLUSION: Within the limits of this in vitro study, rigid custom trays produced significantly more accurate impressions than the polycarbonate stock trays. The stock trays used in this study could not produce accurate impressions consistently. For analogs with a 20-mm separation, there was a difference in medians of 10 microm in accuracy between the stock and custom trays. PMID- 12644800 TI - Fit of implant frameworks: an in vitro comparison between two fabrication techniques. AB - STATEMENT OF PROBLEM: It has been suggested that a precise fit between the implant and the framework cylinder is necessary to ensure a satisfactory long term clinical outcome. PURPOSE: The purpose of this study was to compare the precision of fit between implant abutments and framework cylinders in frameworks fabricated by the Procera system and those fabricated from cast gold-alloy. MATERIAL AND METHODS: A total of 19 frameworks, 14 made with the Procera system (type 1) and 5 made of a cast gold-alloy (type 2), were fabricated. A total of 95 implants, 70 type 1 and 25 type 2 frameworks, were evaluated. Three replicas of the space between the implant abutments and the framework cylinders of the master cast were made for each test specimen. The replicas were cut with a scalpel in 2 axial directions: buccal-lingual and right-left. For the purpose of measurement, a microscope with a precision of +/- 0.5 microm was used at original magnification x30. The Student t test was used to determine whether there were significant differences between the framework designs. RESULTS: The buccal lingual measurements for the type 1 and type 2 frameworks showed mean values of 28.1 microm (SD 9.8) and 42.0 microm (SD 1.8) on the buccal side, respectively, and 25.6 microm (SD 11.2) and 51.6 microm (SD 10.9) on the lingual side, respectively. For the right-left view, the mean measurements were 26.6 microm (SD 8.4) and 49.2 microm (SD 11.4) on the right side, respectively, and 27.4 microm (SD 8.5) and 44.4 microm (SD 6.5) on the left side, respectively. The total mean value for type 1 frameworks was 26.9 microm (SD 9.3); that for type 2 frameworks was 46.8 microm (SD 8.8). CONCLUSION: Within the limitations of this experiment, it was demonstrated that the fit of frameworks made with the Procera system was significantly better than that of the frameworks made with cast gold-alloy (P<.01). PMID- 12644801 TI - Fracture toughness of nine flowable resin composites. AB - STATEMENT OF PROBLEM: Flowable composite is used in a variety of clinical applications; however, the capacity of these materials to resist crack propagation is not well understood. PURPOSE: The purpose of this investigation was to compare the resistance to crack propagation of 9 flowable composites as measured by the fracture toughness. MATERIAL AND METHODS: The composites studied include AeliteFlo, Crystal Essence, Flow-it, FloRestore, Permaflo, Revolution, Tetric Flow, VersaFlo, and Wave. Ten specimens of each composite were formed with a brass mold with a 3-mm preformed notch. The final dimensions of each specimen were 2 x 4.2 x 20 mm. All specimens were light-polymerized to manufacturer specifications and stored in air for 24 hours. The fracture toughness value, K(IC) (MNm(-3/2)), for each specimen was measured by use of a 3-point bending mode and a single-edge notched beam at a crosshead speed of 0.125 mm/min until fracture. The data were analyzed statistically by use of 1-way analysis of variance, t tests (P<.05), and regression analysis. RESULTS: The flowable composites tested showed a spectrum of fracture toughness values ranging from 1.15 +/- 0.10 MNm(-3/2) for Wave to 1.65 +/- 0.13 MNm(-3/2) for Permaflo (significantly different, P<.05). The remaining materials formed 1 group with intermediate K(IC) values not different from each other (P>.05) but significantly different from Wave and Permaflo. Comparisons of fracture toughness to the filler content by volume of each composite revealed no correlation. CONCLUSION: This in vitro study concluded that there was no significant difference among 7 of the 9 composites tested in their resistance to fracture. Permaflo showed the greatest resistance to crack propagation. There was no correlation between the filler content by volume and the fracture toughness of these flowable composites. PMID- 12644802 TI - Resin-ceramic bonding: a review of the literature. AB - Current ceramic materials offer preferred optical properties for highly esthetic restorations. The inherent brittleness of some ceramic materials, specific treatment modalities, and certain clinical situations require resin bonding of the completed ceramic restoration to the supporting tooth structures for long term clinical success. This article presents a literature review on the resin bond to dental ceramics. A PubMed database search was conducted for in vitro studies pertaining to the resin bond to ceramic materials. The search was limited to peer-reviewed articles published in English between 1966 and 2001. Although the resin bond to silica-based ceramics is well researched and documented, few in vitro studies on the resin bond to high-strength ceramic materials were identified. Available data suggest that resin bonding to these materials is less predictable and requires substantially different bonding methods than to silica based ceramics. Further in vitro studies, as well as controlled clinical trials, are needed. PMID- 12644803 TI - Dentin bond strengths of two ceramic inlay systems after cementation with three different techniques and one bonding system. AB - STATEMENT OF PROBLEM: Cementation of inlay restoration is critical. Because of its high organic content, dentin is a less favorable substrate for bonding than enamel. Therefore it is important to improve dentin adhesion when placing ceramic inlay restorations. PURPOSE: The purpose of this study was to compare the dentin bond strengths of 2 different ceramic inlay systems after cementation with 3 different techniques and 1 bonding system. MATERIAL AND METHODS: One hundred twenty freshly extracted caries- and restoration-free molar teeth used in this study were stored in saline solution at room temperature. Standardized Class I preparations were made in all teeth. Each preparation had a length of 6 mm, a width of 3 mm, a depth of 2 mm, and 6-degree convergence of the walls. Teeth were randomly assigned to 2 groups of 60 each to evaluate the bonding of 2 ceramic systems, Ceramco II (Group I) and IPS Empress 2 (Group II), to dentin. Each of the 2 groups were further divided into 3 cementation technique groups of 20 each (Group I A, B, and C and Group II A, B, and C). Groups I A and B and Groups II A and B used dentin bonding agent (DBA) Clearfil Liner Bond 2V, and resin cement (Panavia F). Groups I C and II C served as control groups and used Panavia F without the dentin-bonding agent. In Groups I A and II A, the DBA was applied immediately after the completion of the preparations (D-DBA). Impressions were then made, and the ceramic inlays were fabricated according to the manufacturers' guidelines. In Groups I B and II B the DBA was applied just before luting the inlay restorations (I-DBA). In Groups I C and II C, no bonding agent was used before the cementation of the inlay restorations (No DBA). Cementation procedures followed a standard protocol. After cementation, specimens were stored in distilled water at 37 degrees C for 24 hours. The teeth were sectioned both mesial-distally and buccal-lingually along their long axis into three 1.2 x 1.2 mm wide |-shaped sections. The specimens were then subjected to microtensile testing at a crosshead speed of 1 mm/min, and the maximum load at fracture (in kilograms) was recorded. Two-way analysis of variance and Tukey honestly significant difference tests were used to evaluate the results (P<.05). Scanning electron microscopy analysis was used to examine the details of the bonding interface. The fractured surfaces were observed with a stereomicroscope at original magnification x22 to identify the mode of fracture. RESULTS: Although no significant difference was found among the 2 ceramic systems with regard to dentin bond strengths (P>.05), the difference between the cementation techniques was found to be significant (P<.001). Comparison among techniques showed that the dentin bond strength in the D-DBA technique had a significantly higher mean (40.27 +/- 8.55 Kg) than the I-DBA (30.20 +/- 6.78 Kg) and No DBA techniques (32.43 +/- 8.58 Kg). As a result of scanning electron microscopy analysis, a distinct and thicker hybrid zone with more, and longer resin tags were found in specimens treated with the D-DBA technique than with the other 2 techniques. Most failures (353 of 360) were adhesive in nature at the bonding resin/dentin interface. Only 7 specimens showed cohesive failure within the bonding resin. CONCLUSION: Within the limitations of this in vitro study, the cementation of the ceramic inlays tested with the D-DBA technique used resulted in higher bond strengths to dentin. PMID- 12644804 TI - Observable deviation of the facial and anterior tooth midlines. AB - STATEMENT OF PROBLEM: It has been recommended that the anterior tooth midline be placed coincidently with the midline of the face (facial midline). The location of the facial midline depends largely on the judgment of the clinician. The amount of deviation between these 2 midlines that is noticeable to the observer has not been fully investigated. PURPOSE: The observable deviation between the anterior tooth and facial midlines in a limited sample of dentate subjects was recorded. MATERIAL AND METHODS: Full facial-view, standardized photographs (x1/1.5) of 45 subjects, meeting very limited inclusion/exclusion criteria and undergoing routine dental treatment at Tel Aviv Dental School, were examined by 10 observers: 5 dentists and 5 nondental personnel who were given only a brief explanation of facial and tooth midlines. The examiners asked whether the facial and anterior tooth midlines deviated. The photographs then were scanned onto a computer screen, and the facial midline was determined by bisecting the distance between the medial angles of the eyes. The distance between a line perpendicular to this point and the contact point of the central incisors was measured by one calibrated examiner. The photographs were grouped according to the midline deviation: group 1, <1 mm; group 2, 1 to 2 mm; and group 3, >2 mm. Two photographs with oblique anterior tooth midlines were removed from the study because they were so easily detected. The observers' detection rates for the remaining midline deviation were compared and subjected to 1-way analysis of variance to identify significant differences at the 95% level of confidence. A post hoc Student t test was performed to identify significant differences among the groups. RESULTS: Dentists and nondental personnel demonstrated a similar ability to notice deviations of anterior tooth and facial midlines. Midline deviations of <1 mm (group 1) were detected by 6 of 10 observers in 4 of 29 photographs (14%). Midline deviations of 1 to 2 mm (group 2) were detected by 6 observers in 3 of 8 photographs (37%). Midline deviations of >2 mm (group 3) were detected by 6 observers in 5 of 6 photographs (83%). The greater the deviation, the higher the detection rate. Significant differences were found between group 3 and the other 2 groups (P<.01). CONCLUSION: Within the limitations of this study, the greater the deviation of anterior tooth and facial midlines, the higher the detection rate. Nearly half of the 10 observers involved in this investigation were unable to detect midline deviations of <2 mm. PMID- 12644806 TI - The teaching and treatment of upper airway sleep disorders in North American dental schools. AB - STATEMENT OF PROBLEM: Many dental patients suffer from upper airway sleep disorders (UASD) that affect the quantity and quality of their sleep. These disorders vary from minor annoyances to life-threatening conditions. Even though dentists can play a role in the successful treatment of these patients with oral devices, few do. This lack of participation, in part, may be due to lack of education. PURPOSE: The purpose of this study was to determine which North American dental schools include the treatment of UASD in their curriculum. This information may prove helpful to other schools in determining whether, and to what degree, they should teach about UASD and its treatment options. MATERIAL AND METHODS: A survey instrument (questionnaire) was mailed to all of the North American dental schools (n = 64) affiliated with the American Association of Dental Schools. The questionnaire consisted of 9 multipart questions. Major categories of questions included the following: (1) Is the subject of UASD a part of the school curriculum? If so, in what department is it taught? (2) Are oral devices fabricated for patients or is the experience only at the didactic level? (3) What diagnostic procedures are required? (4) What types of oral devices are fabricated and what is the success rate/recall frequency? (5) What is the patient source? (6) If this subject area is not covered either didactically or clinically, why not? The data were analyzed by use of descriptive statistics. RESULTS: Forty-three of the 64 schools that received the survey responded. Forty two percent of the schools that responded are teaching the treatment of UASD with oral devices. There is no apparent consensus with regard to whether to teach the subject, at what level, or to what degree. CONCLUSION: Even though many dental patients suffer from potentially life-threatening sleep disorders, many of which could be treated with oral devices, fewer than half of the reporting dental schools currently include this area as part of their curriculum. This lack of inclusion is caused primarily by a lack of familiarity with the information available and a lack of available curriculum time. PMID- 12644805 TI - The magnitude of cutting forces at high speed. AB - STATEMENT OF THE PROBLEM: Previous research has used a range of cutting forces for preparing teeth, but there are no data available on the forces actually imposed while cutting teeth with high-speed handpieces. PURPOSE: The purpose of this study was to measure the forces imposed while cutting teeth with tungsten carbide burs used in high-speed handpieces. MATERIAL AND METHODS: Thirty-one dentists each cut 8 conventional class II MO and DO preparations in intact extracted third molars, by use of 2 different air turbine handpieces with different torque-speed characteristics. Two different flat fissure, plain and cross-cut tungsten carbide burs, cutting wet and dry in each handpiece/bur combination. The teeth were mounted in a custom-made transducer unit that displayed the forces imposed by the bur. Data were analysed with a 1-way ANOVA (alpha =.05) and Spearman correlation test. RESULTS: The results showed that there was no significant difference in the applied force between plain and cross cut burs, cutting wet or cutting dry, but there was a significant difference between the high and the lower torque handpieces. The higher torque handpiece was used at a mean cutting force of 1.44 N and the lower torque handpiece at 1.20 N (P<.002). The overall general mean force observed was 1.30 N. CONCLUSION: It was concluded that the forces used in cutting teeth with the tungsten carbide burs tested related both to the type of the handpiece and to the forces chosen by clinical operators. There was no difference between the plain and cross-cut burs, there was no difference between the cutting wet or dry, and the higher torque handpiece required a higher mean cutting force. PMID- 12644807 TI - M. M. House mental classification revisited: Intersection of particular patient types and particular dentist's needs. AB - Fifty-three years ago, M. M. House devised a classification of patients on the basis of how they behaved in response to the prospect of becoming edentulous and on how they subsequently adapted to wearing complete dentures. Although House's system was an important contribution, it failed to consider the dentist's emotional reaction to a patient's behavior as part of an understanding of how the patient and doctor cope with the dental treatment. This article presents an expansion of the House classification to include the behavior of the dentist as a codeterminer of the patient's behavior. This expanded classification system is based on empiricism and awaits scientific validation or clinical application to determine its ultimate validity, reliability, and effectiveness. PMID- 12644808 TI - Effect of thermocycling on tensile bond strength of six silicone-based, resilient denture liners. AB - STATEMENT OF PROBLEM: Adhesion failure between silicone resilient denture lining materials and denture base resin is commonly encountered in clinical practice. Adhesion failure results in localized unhygienic conditions at the debonded regions and often causes functional failure of the prosthesis. PURPOSE: The purpose of this study was to investigate the effect of thermocycling on the tensile bond strength of 6 soft lining materials. MATERIAL AND METHODS: Six commonly used silicone-based resilient denture liners (Ufigel C, Ufigel P, Mollosil, Molloplast B, Permafix, and Permaflex) were chosen for the investigation. The bond strength was determined, in tension, after processing to PMMA. The resilient denture liners for each group (n = 24) were 10 x 10 x 3 mm and were processed between 2 polymethyl methacrylate specimens according to manufacturer's instructions. Two PMMA specimens were prepared by investing brass dies with a 3-mm-thick spacer in a denture flask. Specimens were made by processing the resilient denture liners against the polymerized PMMA block. After polymerization, the brass spacer was removed from the mold, the 2 PMMA resin specimens were trimmed, and the surfaces to be bonded were smoothed. The PMMA block was placed back into the molds and the resilient denture liners were packed into the space made by brass spacer, trial packed, and polymerized according to the manufacturer's instructions. Half of the specimens for each group were stored in water for 24 hours, and the other half were thermocycled (5000 cycles) between baths of 5 degrees and 55 degrees C. All specimens were placed under tension until failure in a universal testing machine at a crosshead speed of 5 mm/min. The maximum tensile stress before failure and mode of failure were recorded. The mode of failure was characterized as cohesive, adhesive, or mixed mode, depending on whether the fracture surface was in the soft liner only, at the denture base soft liner interface only, or in both. Failure strength was recorded in kg/cm(2). Results were tested by multiple analysis of variance (ANOVA) for mode of failure (adhesive, cohesive, and mix), 2-way ANOVA (storage-products), and 1-way ANOVA (storage-products interaction, before and after thermocycling). Duncan's test was used to determine whether significant changes in the tensile bond properties of the materials occurred during thermocycling. RESULTS: Varying degrees of bond strengths were found for soft lining materials and were significantly different (P<.05). Results of this study also indicated that the bond strengths of soft lining materials had significantly decreased after thermocycling except Ufigel C and Mollosil. CONCLUSIONS: Because the adequate adhesive value for soft lining materials is given 4.5 kg/cm(2), all of the materials were acceptable for clinical use. PMID- 12644809 TI - Sound analysis of temporomandibular joint internal derangements with phonographic recordings. AB - STATEMENT OF PROBLEM: Temporomandibular joint (TMJ) sound recordings could be analyzed to assess the state of TMJ internal derangements. PURPOSE: The aim of the study was to assess the value of sound analysis in the diagnosis of the type of the TMJ internal derangements. MATERIAL AND METHODS: After clinical and radiologic examinations, phonographic sound recordings on mandibular excursions were obtained in 52 patients with TMJ internal derangements and 12 control individuals. Sound correlations were made on the basis of opening-closing, protrusive-retrusive, and lateral excursions of the mandible. RESULTS: Clicking was a consistent finding of anterior disc displacement with reduction, whereas crepitation was found in varying degrees in anterior disc displacement and osteodegenerative arthritis. Silent TMJs were the feature of normal TMJs, except for the situations of acute lock. Although in 29 TMJs opening click was followed by a closing click (reciprocal clicking), 46 TMJs with opening click also had clicking on protrusion. On the other hand, 19 TMJs with opening click also had clicking on ipsilateral motion, and 40 TMJs with opening click had clicking on contralateral motion of the mandible. The sound patterns were found to be similar in opening-protrusive clicks and opening-contralateral clicks. The lack of protrusive clicking in the presence of opening click was considered an indication of late disc reduction on opening. Crepitation was observed in advanced cases of TMJ internal derangements. CONCLUSION: Within the limitations of this study, the results suggest that TMJ sound analysis on mandibular excursions was indicative for diagnosis and establishment of severity of TMJ internal derangements. Clicking and crepitation may be looked on as signs of abnormal joint disorder, clicking indicating anterior disc displacement with reduction, and crepitation, indicating progression from anterior disc displacement without reduction to osteodegenerative arthritis. PMID- 12644810 TI - Expedient direct approach for esthetic and functional provisional restorations. AB - This article describes a direct procedure for the expeditious fabrication of provisional restorations for extensive fixed prosthodontic applications. Customary indirect or indirect-direct approaches require extensive clinical time as well as laboratory support for their successful application. Provided that the necessary preoperative steps are followed precisely, this treatment method allows for excellent clinical results while being less time consuming. PMID- 12644811 TI - A handle for facial casts. PMID- 12644812 TI - Nondestructive examination of all-ceramic fixed partial denture with microcomputed tomography. PMID- 12644814 TI - Phase I/II trial of weekly cisplatin, etoposide, and irinotecan chemotherapy for metastatic lung cancer: JCOG 9507. AB - Combinations of cisplatin-irinotecan and cisplatin-etoposide are active and well tolerated in patients with both small-cell lung cancer (SCLC) and nonsmall-cell lung cancer (NSCLC). To define the recommended dose for phase II trials of irinotecan combined with cisplatin and etoposide in chemonaive patients with stage IV disease, 56 patients (11 having SCLC and 45 NSCLC) received cisplatin 25 mg m(-2) weekly for 9 weeks, etoposide 60 mg m(-2) for 3 days on weeks 1, 3, 5, 7 and 9, and irinotecan 20-100 mg m(-2) (levels 1-8) on weeks 2, 4, 6 and 8, together with a prophylactical granulocyte colony-stimulating factor support (50 microg m(-2) on days 4-7 on weeks 1, 3, 5, 7 and 9, and on days 2-7 on weeks 2, 4, 6 and 8). Grade 3-4 leukocytopenia, neutropenia and thrombocytopenia were noted in 20 (36%), 28 (50%) and nine (16%) patients, respectively. Grade 3 diarrhoea, grade 3 cardiac toxicity, and grade 4 transaminase elevation developed in one (1.8%) patient each. Totally, four of 56 patients were removed from the study because of toxicity and recovered, and two other patients died in situations where drug toxicity might contribute to their death. Dose-limiting toxicity was noted in less than one-third of patients at dose levels 1-7, but in all patients at dose level 8. Thus, the recommended dose was determined to be level 7 (irinotecan 90 mg m(-2)). The response rates for SCLC and NSCLC were 91% (10/11) and 38% (17/45), respectively. The median survival time and 1-year survival rate were 11.9 months and 46% for SCLC and 10.1 months and 40% for NSCLC, respectively. This regimen was considered to be feasible and promising for the treatment of stage IV SCLC and NSCLC. PMID- 12644813 TI - Nonsteroidal anti-inflammatory drugs in colorectal cancer: from prevention to therapy. AB - In this review, we discuss the available experimental evidences supporting the chemopreventive efficacy of nonsteroidal anti-inflammatory drugs (NSAIDs) on colorectal cancer and the biological basis for their possible role as anticancer agents. Although the comprehension of the mechanisms underlying the effects of these drugs on colon cancer cells is incomplete, research efforts in identifying the biochemical pathway by which NSAIDs exert their chemopreventive effect have provided a rationale for the potential use of NSAIDs alone or in combination with conventional and experimental anticancer agents in the treatment of colorectal cancer. In this paper, we review three main issues: (i) the role of COX-2 in colon cancer; (ii) the common death pathways between NSAIDs and anticancer drugs; and (iii) the biological basis for the combination therapy with COX-2 selective inhibitors and new selective inhibitors of growth factor signal transduction pathways. PMID- 12644815 TI - Adaptive intrapatient dose escalation of cisplatin in combination with low-dose vp16 in patients with nonsmall cell lung cancer. AB - The objective of this phase II and pharmacologic study was to explore the feasibility, toxicity and activity of adaptive intrapatient dose escalation of cisplatin in a dose-intensive weekly schedule using predefined levels of exposure, with the ultimate aim to improve the antitumour activity of the therapy in patients with nonsmall cell lung cancer (NSCLC). Platinum DNA-adduct levels in peripheral white blood cells during treatment were used as the primary parameter for adaptive dosing. If DNA-adduct levels were not available, the area under the concentration-time curve (AUC) of unbound platinum in plasma was used for dose adaptation. Target levels for DNA-adducts and AUC have been defined in a previously performed pharmacologic study. The feasibility of adaptive dosing was tested in 76 patients with stage IIIB and IV NSCLC, who were planned to receive 6 weekly courses of cisplatin at a starting dose of 70 mg m(-2), together with daily low oral dose of 50 mg VP16. In total, 37 patients (49%) who were given more than one course received a dose increase varying from 10 to 55%. The majority of patients reached the defined target levels by a dose increase during course two. Relevant grade 2 neurotoxicity was observed in eight (10%) patients and reversible ototoxicity grade 2 in 14 (18%) patients. The strategy of adaptive intrapatient dose adjustment of cisplatin is practically feasible in a research setting even when results for dose adaptation have to be reported within a short time-period of 1 week. The toxicity appeared to be manageable in this cohort of patients. In some patients, exposure after the standard dose was substantially lower than the defined target level and significant dose escalations of more than 50% had to be applied. The response rate (RR) was relatively high: overall 40% (29 out of 72 patients) partial remission (PR), in patients with stage IIIB the RR was 60% (15 out of 25 patients) and with stage IV 30% (14 out of 47 patients). Randomised studies are needed to determine whether the adaptive dosing strategy results in better efficacy than standard dosing. PMID- 12644816 TI - An open-label phase II study of low-dose thalidomide in androgen-independent prostate cancer. AB - The antiangiogenic effects of thalidomide have been assessed in clinical trials in patients with various solid and haematological malignancies. Thalidomide blocks the activity of angiogenic agents including bFGF, VEGF and IL-6. We undertook an open-label study using thalidomide 100 mg once daily for up to 6 months in 20 men with androgen-independent prostate cancer. The mean time of study was 109 days (median 107, range 4-184 days). Patients underwent regular measurement of prostate-specific antigen (PSA), urea and electrolytes, serum bFGF and VEGF. Three men (15%) showed a decline in serum PSA of at least 50%, sustained throughout treatment. Of 16 men treated for at least 2 months, six (37.5%) showed a fall in absolute PSA by a median of 48%. Increasing levels of serum bFGF and VEGF were associated with progressive disease; five of six men who demonstrated a fall in PSA also showed a decline in bFGF and VEGF levels, and three of four men with a rising PSA showed an increase in both growth factors. Adverse effects included constipation, morning drowsiness, dizziness and rash, and resulted in withdrawal from the study by three men. Evidence of peripheral sensory neuropathy was found in nine of 13 men before treatment. In the seven men who completed six months on thalidomide, subclinical evidence of peripheral neuropathy was found in four before treatment, but in all seven at repeat testing. The findings indicate that thalidomide may be an option for patients who have failed other forms of therapy, provided close follow-up is maintained for development of peripheral neuropathy. PMID- 12644817 TI - Radiotherapy with 16 Gy may fail to eradicate testicular intraepithelial neoplasia: preliminary communication of a dose-reduction trial of the German Testicular Cancer Study Group. AB - Low-dose radiotherapy to the testis is effective in eradicating testicular intraepithelial neoplasia (TIN, carcinoma in situ of the testis) at the risk of androgenic deficiency. The present trial was designed to define the lowest dose effective to control TIN assuming a dose-response relation of radiation-induced endocrinological damage. Patients with TIN in a solitary testicle or with bilateral TIN were treated with 18 Gy (14 patients) and 16 Gy (26 patients) (5 x 2 Gy per week). Biopsies to ascertain clearance of TIN were performed after 6 and 24 months. The median time of follow-up is 20.5 months. There were three adverse events. In one patient, relapse of TIN along with microinvasive seminoma was observed 2 years after 16 Gy irradiation. In two other patients, persistent spermatogonia were observed with the 16 and 18 Gy regimen after 6 and 24 months, respectively. All other post-treatment biopsies showed the Sertoli cell-only pattern. These results confirm that TIN is a radiosensitive lesion efficiently controlled in most cases with doses below 20 Gy. However, sporadic failures may occur. A dose of 16 Gy is probably unsafe and should no longer be used. Future investigations should not only focus on total dosage of irradiation but also on fractionation schedules. PMID- 12644818 TI - Stage of breast cancer at diagnosis among women with cosmetic breast implants. AB - Concern has been raised about the potential delay in breast cancer diagnosis in the augmented breast. We linked a cohort of 2955 women, who received cosmetic breast implants in Denmark during the period 1973-1997 with the Danish Cancer Registry and the Danish Breast Cancer Cooperative Group register. We identified 23 incident cases of invasive breast cancer diagnosed subsequent to breast implantation. We randomly selected 11 controls for each case from the Danish Breast Cancer Cooperative Group's register, and obtained detailed information on all study subjects about surgery, histopathology and stage of breast cancer at diagnosis, intended adjuvant treatment according to trial protocols and overall survival. We found that women with breast implants on average were diagnosed with breast cancer at the same stage as controls. Significantly more women with breast implants had tumour cells in the surgical margins according to the Danish Breast Cancer Cooperative Group's data. There was no significant difference in overall survival between the two groups after an average of 6.4 years of follow-up. Based on this limited number of women with breast cancer subsequent to breast augmentation, breast implants do not appear to delay the diagnosis of breast cancer, and no evidence of impaired survival after breast cancer diagnosis in augmented women was found. PMID- 12644819 TI - How doctors record breaking bad news in ovarian cancer. AB - Revealing the diagnosis of cancer to patients is a key event in their cancer journey. At present, there are no minimal legal recommendations for documenting such consultations. We reviewed the Hospital records of 359 patients with epithelial ovarian cancer in the Mersey Area between 1992 and 1994. We identified the following factors: age, hospital, postcode, surgeon, stage of disease and survival. These were compared to information recorded at the time of the interview such as person present, descriptive words used, prognosis, further treatment and emotional response. In 11.6%, there was no information recorded in the notes. The diagnosis was recorded in 304 (94.7%), prognosis in 66 (20.6%) and collusion with relatives in 33 (10.3%). A total of 42 separate words/phrases were identified relating to diagnosis; cancer was recorded in 60 (19.6%). Collusion was three times as common in the patients over 65 years (17.9 vs 5.7%, P=0.001). There was a reduction in the number of diagnostic words recorded in the patients over 65 years (90.3 vs 98.3%, P=0.002) and by type of surgeon (P=0.001). Information was often poorly recorded in the notes. We have shown that the quality of information varies according to patient age, surgeon and specialty. PMID- 12644820 TI - External validity of a prediction rule for residual mass histology in testicular cancer: an evaluation for good prognosis patients. AB - We assessed the external validity of a prediction rule for nonseminomatous testicular cancer patients. The rule was developed to predict the probability of retroperitoneal metastases being benign (only necrosis/fibrosis) after chemotherapy treatment. Patients with a high probability of benign residual masses might be offered surveillance as opposed to patients with a low probability, who should undergo retroperitoneal lymph node dissection (RPLND). We compared the observed histology with the predicted probability in 105 patients with good prognosis germ cell cancer who underwent RPLND between 1995 and 1998. We found that predicted probabilities higher than 5% were in good agreement with the observed frequencies of benign masses. The area under the receiver operating characteristic curve was 0.76, suggesting that the rule could reasonably discriminate between benign masses and tumour. However, nearly all predicted probabilities (n=101) were lower than 70%, which might be considered as the lowest value at which surveillance offers a reasonable alternative to RPLND. Further, 35% of patients currently under surveillance (84 out of 241) had predicted probabilities lower than 70%. In conclusion, the clinical relevance of the prediction rule was limited for the patients who underwent RPLND; use of the rule would change the policy from RPLND to surveillance in only a few. On the other hand, the rule might support selection of patients for RPLND, who currently are under surveillance. PMID- 12644822 TI - Chemokine receptor CCR2 is expressed by human multiple myeloma cells and mediates migration to bone marrow stromal cell-produced monocyte chemotactic proteins MCP 1, -2 and -3. AB - The restricted bone marrow (BM) localisation of multiple myeloma (MM) cells most likely results from a specific homing influenced by chemotactic factors, combined with the proper signals for growth and survival provided by the BM microenvironment. In analogy to the migration and homing of normal lymphocytes, one can hypothesise that the BM homing of MM cells is mediated by a multistep process, involving the concerted action of adhesion molecules and chemokines. In this study, we report that primary MM cells and myeloma derived cell lines (Karpas, LP-1 and MM5.1) express the chemokine receptor CCR2. In addition, we found that the monocyte chemotactic proteins (MCPs) MCP-1, -2 and -3, three chemokines acting as prominent ligands for CCR2, are produced by stromal cells, cultured from normal and MM BM samples. Conditioned medium (CM) from BM stromal cells, as well as MCP-1, -2 and -3, act as chemoattractants for human MM cells. Moreover, a blocking antibody against CCR2, as well as a combination of neutralizing antibodies against MCP-1, -2 and -3, significantly reduced the migration of human MM cells to BM stromal cell CM. The results obtained in this study indicate the involvement of CCR2 and the MCPs in the BM homing of human MM cells. PMID- 12644821 TI - Evaluation of clinical significance of TP53, BCL-2, BAX and MEK1 expression in 229 ovarian carcinomas treated with platinum-based regimen. AB - In cell line studies, BCL-2, BAX, as well as novel MEK1 protein levels have strong influence on ovarian cancer response to cisplatin-based chemotherapy. However, such associations have not been demonstrated clinically. We evaluated prognostic/predictive significance of these proteins with regard to TP53 status. Immunohistochemical analysis was performed on 229 ovarian carcinomas FIGO stage IIB-IV treated with platinum-based chemotherapy; the results were analysed by the Cox and logistic regression models. Clinical parameters (residual tumour size, patient age, FIGO stage) were the only indicators of overall survival (OS) and the strongest predictors of complete remission (CR). On the other hand, BAX expression was the strongest (P=0.005) or the only (in FIGO IIIC, P=0.02) prognostic indicator of disease-free survival (DFS) in the TP53(+) group. TP53(+) and TP53(-) ovarian carcinomas differed in clinical and molecular prognostic and predictive factors. Another novel finding is that CR was negatively influenced by high BAX expression in all patients group (P=0.047) and by BCL2 expression in the TP53(-) group (P=0.05). High MEK1 expression was associated with endometrioid and clear cell carcinomas (P=0.049); its loss was found with advancing FIGO stage (P=0.002). Our results suggest that binomial TP53 status divides ovarian carcinomas into two biologically distinct groups. BAX expression is an important factor of DFS in the TP53(+) group. BCL-2 and BAX, but not MEK1 expressions have predictive value in ovarian cancer patients treated with platinum-based chemotherapy. PMID- 12644823 TI - Prognostic value of maspin mRNA expression in ER alpha-positive postmenopausal breast carcinomas. AB - Maspin, a member of the serpin family, has a role in cell migration, angiogenesis and apoptosis. Little is known of the clinical significance of maspin gene expression in human cancers. We developed a real-time quantitative RT-PCR assay to quantify the full range of maspin mRNA copy numbers in a series of 10 ER alpha positive and 10 ER alpha-negative breast tumours. We observed a statistical link between low maspin mRNA levels and positive oestrogen status (P=0.0012). In consequence, to better assess the prognostic value of maspin gene expression in breast cancer, we then quantified maspin mRNA content in an additional independent well-defined cohort of 105 ER alpha-positive postmenopausal breast cancer patients treated with primary surgery followed by adjuvant tamoxifen alone. Maspin expression varied widely in tumour tissues (by nearly four orders of magnitude), being underexpressed in 33 out of 105 tumours (31.4%) and overexpressed in 24 out of 105 tumours (22.9%) relative to normal breast tissues. Immunohistochemical studies demonstrated that maspin protein was strictly expressed in myoepithelial cells of normal breast tissue and in tumour epithelial cells, exclusively in maspin-overexpressing tumours. Patients with tumours overexpressing the maspin gene had significantly shorter relapse-free survival after surgery than patients whose tumours normally expressed or underexpressed maspin (P=0.0011). The prognostic significance of maspin overexpression persisted in Cox multivariate regression analysis (P=0.0024). These findings show that the maspin mRNA level can have important prognostic significance in human breast cancer, and point to the maspin gene as a putative molecular predictor of hormone responsiveness in breast cancer. PMID- 12644824 TI - Elevated expression and altered processing of fibulin-1 protein in human breast cancer. AB - The extracellular matrix protein fibulin-1 suppresses the motility and invasiveness of a variety of tumour cell types in vitro as well as the growth of fibrosarcoma tumours in nude mice. In this study, fibulin-1 protein expression in breast carcinoma specimens and normal breast tissue was evaluated immunohistologically. Fibulin-1 protein expression was also semiquantitatively assessed by immunoblot analysis in a collection of normal breast tissues (n=18), benign tumours (n=5) and breast carcinomas (n=39). In normal breast tissue, fibulin-1 protein expression predominated in the ductal epithelium and underlying myoepithelium, with weaker staining evident in the loose connective surrounding the ducts. Examination of breast carcinomas revealed that the tumour cells also expressed fibulin-1 protein. The level of mature fibulin-1 polypeptide (100 kDa) was higher in the breast carcinoma specimens as compared to normal breast tissue (Mann-Whitney U-test, P=0.0005). In addition to the mature fibulin-1 polypeptide, several smaller sized polypeptides of 55, 50 and 25 kDa were detected using monoclonal antibodies reactive towards an epitope located at the N-terminus of fibulin-1. The immunoreactive 50 kDa polypeptide was detected more frequently in breast carcinoma specimens than in normal breast tissue (chi(2)=17.22, P<0.0001). Furthermore, the ratio of the 50 kDa fragment to the mature fibulin-1 polypeptide correlated with the level of oestrogen receptor alpha (Spearman correlation coefficient, rs=0.49, P<0.003, n=36) and progesterone receptor (rs=0.43, P=0.008, n=36) expression in the tumour specimens. Taken together, these findings indicate that elevated expression and altered processing of fibulin-1 is associated with human breast cancer. PMID- 12644826 TI - L523S, an RNA-binding protein as a potential therapeutic target for lung cancer. AB - Approaches to vaccine-based immunotherapy of human cancer may ultimately require targets that are both tumour-specific and immunogenic. In order to generate specific antitumour immune responses to lung cancer, we have sought lung cancer specific proteins that can be targeted for adjuvant vaccine therapy. By using a combination of cDNA subtraction and microarray analysis, we previously reported the identification of an RNA-binding protein within the KOC family, L523S, to be overexpressed in squamous cell cancers of the lung. We show here that L523S exhibits significant potential for vaccine immunotherapy of lung cancer. As an oncofetal protein, L523S is normally expressed in early embryonic tissues, yet it is re-expressed in a high percentage of nonsmall cell lung carcinoma. The specificity of L523S expression in lung cancer was demonstrated by both mRNA and protein measurements using real-time PCR, Western blot, and immunohistochemistry analyses. Furthermore, we show that immunological tolerance of L523S is naturally broken in lung cancer patients, as evidenced by detectable antibody responses to recombinant L523S protein in eight of 17 lung pleural effusions from lung cancer patients. Collectively, our studies suggest that L523S may be an important marker of malignant progression in human lung cancer, and further suggest that treatment approaches based on L523S as an immunogenic target are worthy of pursuit. PMID- 12644825 TI - Lung resistance-related protein as a predictor of clinical outcome in advanced testicular germ-cell tumours. AB - This study was undertaken to investigate the expression and predictive value for outcome of multidrug resistance-associated (MDR) proteins P-glycoprotein (Pgp), MRP1, BCRP, and LRP, in advanced testicular germ-cell tumours (TGCT). Paraffin embedded sections from 56 previously untreated patients with metastatic TGCT were immunostained for Pgp, MRP1, BCRP, and LRP. All patients received platinum-based chemotherapy after orchidectomy. Immunostaining was related to clinicopathological parameters, response to chemotherapy, and outcome. Strong and intermediate expressions of the different MDR-related proteins were: 27 and 41% (Pgp), 54 and 37% (MRP1), 86 and 7% (BCRP), and 14 and 29% (LRP). P-glycoprotein and MRP1 associated, respectively, to low AFP (P=0.026) and high LDH levels (P=0.014), whereas LRP expression associated with high beta-hCG levels (P=0.003) and stage IV tumours (P=0.029). No correlation was found between Pgp, MRP1, and BCRP expression and response to chemotherapy and survival. In contrast, patients with LRP-positive tumours (strong or intermediate expression) had shorter progression-free (P=0.0006) and overall survival (P=0.0116) than LRP-negative patients, even after individual log-rank adjustments by statistically associated variables. Our data suggest that a positive LRP immunostaining at the time of diagnosis in metastatic TGCT is associated with an adverse clinical outcome. PMID- 12644827 TI - Detection of replicative integrity in small colonic biopsies using the BrdUrd comet assay. AB - The alkaline single-cell gel electrophoresis or comet assay is a relatively simple method of measuring DNA single-strand breaks and alkali-labile sites in individual cells. Previously, we have used a combination of this with bromodeoxyuridine labelling of DNA and immunolocalisation of the BrdUrd to show that DNA replicative integrity can be assessed in single cultured cells. This study demonstrates the application of the technique to single cells derived from small human colonic biopsies isolated at routine endoscopy. A high level of reproducibility within replicate comet slides and between comet slides prepared from various colonic sites within a single patient is shown. Preliminary results demonstrate that defects in replication can be detected in tumour and premalignant colonic tissue adjacent to the tumour, suggesting that alterations in replicative integrity are an early event in neoplasia, appearing in premalignant mucosal cells. This development deems the BrdUrd comet assay suitable as an ex vivo molecular end point that can be measured easily in tissue collected by biopsy at routine colonic endoscopy. Thus, the BrdUrd comet assay has the potential to facilitate trial investigations of diet- or environment related factors that may affect replicative integrity in the colon and provides a novel biomarker for colon carcinogenesis. PMID- 12644828 TI - Antiapoptotic effect of haem oxygenase-1 induced by nitric oxide in experimental solid tumour. AB - Induction of haem oxygenase-1 (HO-1) may provide an important protective effect for cells against oxidative stress. Here, we investigated the mechanism of cytoprotection of HO-1 in solid tumour with a focus on the antiapoptotic activity of HO-1. Treatment of rat hepatoma AH136B cells with the HO inhibitor zinc protoporphyrin IX (ZnPP IX) or tin protoporphyrin IX resulted in extensive apoptotic changes of tumour cells both in vivo and in vitro. Caspase-3 activity of the ZnPP IX-treated hepatoma cells increased significantly. Moreover, ZnPP IX induced apoptosis was completely inhibited by simultaneous incubation with a specific caspase-3 inhibitor and was partially abrogated by bilirubin, a reaction product of HO. In vivo ZnPP IX treatment did not affect nitric oxide (NO) production and tumour blood flow. Western blot analyses showed that HO-1 expression in AH136B cells was strongly upregulated by NO donors, for example, S nitroso-N-acetyl penicillamine and propylamine NONOate in vitro; conversely, it was remarkably reduced in vivo by pharmacological blockade of NOS. We conclude that HO-1 may function in antiapoptotic defense of the tumour, and thus it may have important protective and beneficial effects for tumour cells against oxidative stress induced by NO, which is produced in excess during solid tumour growth in vivo. PMID- 12644829 TI - Functional screening of genes suppressing TRAIL-induced apoptosis: distinct inhibitory activities of Bcl-XL and Bcl-2. AB - TNF-related apoptosis-inducing ligand (TRAIL) is known to selectively induce apoptosis in various tumour cells. However, downstream-signalling of TRAIL receptor is not well defined. A functional genetic screening was performed to isolate genes interfering with TRAIL-induced apoptosis using cDNA retroviral library. Bcl-X(L) and FLIP were identified after DNA sequencing analysis of cDNA rescued from TRAIL-resistant clones. We found that increased expression of Bcl X(L), but not Bcl-2, suppressed TRAIL-induced apoptosis in tumour cells. Western blot and immunohistochemical analyses showed that expression of Bcl-X(L), but not Bcl-2, was highly increased in human breast cancer tissues. Exposure of MDA-MB 231 breast tumour cells to TRAIL induced apoptosis accompanied by dissipation of mitochondrial membrane potential and enzymatic activation of caspase-3, -8, and 9. However, SK-BR-3 breast tumour cells exhibiting increased expression level of Bcl-X(L) were resistant to TRAIL, though upon exposure to TRAIL, caspase-8 and Bid were activated. Forced expression of Bcl-X(L), but not Bcl-2, desensitised TRAIL-sensitive MDA-MB-231 cells to TRAIL. Similar inhibitory effects were also observed in other tumour cells such as HeLa and Jurkat cells stably expressing Bcl-X(L), but not Bcl-2. These results are indicative of the crucial and distinct function of Bcl-X(L) and Bcl-2 in the modulation of TRAIL-induced apoptosis. PMID- 12644830 TI - TRAIL-beta and TRAIL-gamma: two novel splice variants of the human TNF-related apoptosis-inducing ligand (TRAIL) without apoptotic potential. AB - Tumour necrosis factor (TNF) related apoptosis-inducing ligand (TRAIL/APO2L) is a recently identified member of the TNF family, which induces programmed cell death in a variety of neoplastic cell types, but not in most nonneoplastic cells. In this study, we report on the identification of two novel alternative splice variants of TRAIL in neoplastic and non-neoplastic human cells lacking either exon 3 (TRAIL-beta) or exons 2 and 3 (TRAIL-gamma). In both splice variants, loss of exon 3 resulted in a frame shift generating a stop codon with consecutive extensive truncation in the extracellular domain. Ectopic expression revealed a loss of proapoptotic potential for both alternative splice variants. In contrast to the predominantly cytoplasmatic localisation of GFP-tagged TRAIL-alpha and TRAIL-beta, TRAIL-gamma showed an additional association with the cell surface and nuclear membrane. In conclusion, alternative splicing might be involved in fine tuning of TRAIL-induced apoptosis and underlines the complexity of the TRAIL system. PMID- 12644831 TI - CYP3A4 and VDR gene polymorphisms and the risk of prostate cancer in men with benign prostate hyperplasia. AB - Prostate cancer (PRCa) is one of the most common causes of cancer death in men and determinants of PRCa risk remain largely unidentified. Benign prostatic hyperplasia (BPH) is found in the majority of ageing men and has been associated with PRCa. Many candidate genes have been suggested to be involved in PRCa, such as those that are central to cellular growth and differentiation in the prostate gland. The vitamin D receptor (VDR) and CYP3A4 have been shown to be involved in the regulation of cell proliferation and differentiation in prostate cells. Genetic variations of these genes have been associated with PRCa in case-control studies and may be useful to detect BPH patients that have a higher risk of developing PRCa. The association between CYP3A4 and VDR TaqI SNPs and the risk of developing PRCa have been investigated in this study by determining the variant genotype frequencies of both SNPs in 400 patients with BPH who have been followed clinically for a median of 11 years. The results of this study showed that the incidence rate of PRCa was higher in BPH patients having CYP3A4 variant genotype compared to those with wild type (relative risk (RR)=2.7; 95% CI=0.77-7.66). No association between variant genotype and risk of developing PRCa was observed with the VDR TaqI variant genotype. In addition, the results of combined genotype analysis of these two SNPs showed a borderline significant association between CYP3A4 and VDR TaqI combined variant genotypes and PRCa risk (RR=3.43; 95% CI=0.99-11.77). While independent confirmation is required in further studies, these results provide a potential tool to assist prediction strategies for this important disease. PMID- 12644832 TI - CYP17 promoter polymorphism and breast cancer risk in males and females in relation to BRCA2 status. AB - A T-C polymorphism in the promoter region of the CYP17 gene has been associated with male and female breast cancer risk as well as early-onset familial breast cancer. The potential role of this polymorphism was investigated in relation to breast cancer risk in Icelandic male and female carriers and noncarriers of a BRCA2 mutation. The study population consisted of 39 male and 523 female breast cancer cases and 309 male and 395 female controls. Of the cases, 15 males and 55 females carried a BRCA2 mutation. We did not find a significant association between male breast cancer risk and CYP17 genotypes. Among male breast cancer cases, the frequency of the CC genotype was higher among carriers of the 999del5 mutation (33.3%) than noncarriers (16.7%), although this difference also did not reach a statistical significance. No association was observed with breast cancer risk among females irrespective of menopausal status, stage of the disease or BRCA2 status. Our findings do not indicate a role for the CYP17 T-C polymorphism in female breast cancer, but a role in male carriers of a BRCA2 mutation could not be excluded because of the small sample size. PMID- 12644833 TI - A33scFv-cytosine deaminase: a recombinant protein construct for antibody-directed enzyme-prodrug therapy. AB - A recombinant fusion protein of colon carcinoma binding A33 single chain antibody with cytosine deaminase displayed specific antigen binding and enzyme activity in surface plasmon resonance and is catalytic activity assay. In vitro, it selectively increased the toxicity of 5-FC to A33 antigen-positive cells by 300 fold, demonstrating the potency of this ADEPT strategy. PMID- 12644834 TI - Tumoricidal activity of lauryl gallate towards chemically induced skin tumours in mice. AB - Lauryl gallate (antioxidant food additive E-312) prevents the formation of dimethylbenzanthracene-induced skin tumours in mice, and kills, selectively, tumoral cells on established tumours. This results in total remission, after topical application of the compound on the tumoral mass, without affecting the surrounding tissue. PMID- 12644835 TI - Preclinical studies of targeted alpha therapy for breast cancer using 213Bi labelled-plasminogen activator inhibitor type 2. AB - The control of micrometastatic breast cancer remains problematic. To this end, we are developing a new adjuvant therapy based on (213)Bi-PAI2, in which an alpha emitting nuclide ((213)Bi) is chelated to the plasminogen activator inhibitor-2 (PAI2). PAI2 targets the cell-surface receptor bound urokinase plasminogen activator (uPA), which is involved with the metastatic spread of cancer cells. We have successfully labelled and tested recombinant human PAI2 with the alpha radioisotope (213)Bi to produce (213)Bi-PAI2, which is highly cytotoxic towards breast cancer cell lines. In this study, the 2-day postinoculation model, using MDA-MB-231 breast cancer cells, was shown to be representative of micrometastatic disease. Our in vivo efficacy experiments show that a single local injection of (213)Bi-PAI2 can completely inhibit the growth of tumour at 2 days postcell inoculation, and a single systemic (i.p.) administration at 2 days causes tumour growth inhibition in a dose-dependent manner. The specific role of uPA as the target for (213)Bi-PAI2 therapy was determined by PAI2 pretreatment blocking studies. In vivo toxicity studies in nude mice indicate that up to 100 microCi of (213)Bi-PAI2 is well tolerated. Thus, (213)Bi-PAI2 is successful in targeting isolated breast cancer cells and preangiogenic cell clusters. These results indicate the promising potential of (213)Bi-PAI2 as a novel therapeutic agent for micrometastatic breast cancer. PMID- 12644836 TI - Role of cystine transport in intracellular glutathione level and cisplatin resistance in human ovarian cancer cell lines. AB - Transport system x(c)(-) is a member of plasma membrane heterodimeric amino-acid transporters and consists of two protein components, xCT and 4F2hc. This system mediates cystine entry coupled with the exodus of intracellular glutamate and regulates the intracellular glutathione (GSH) levels in most mammalian cultured cells. We studied the activity of system x(c)(-) and GSH content in human ovarian cancer cell line (A2780) and its cisplatin (CDDP)-resistant variant (A2780DDP). The rate of cystine uptake was approximately 4.5-fold higher in A2780DDP cells than in A2780 cells and the cystine uptake in A2780DDP cells was mediated by system x(c)(-). Intracellular GSH content was much higher in A2780DDP cells but it fell drastically in the presence of excess glutamate, which inhibited the cystine uptake competitively. xCT and 4F2hc mRNAs were definitely expressed in A2780DDP cells, but far less in A2780 cells. Expression of system x(c)(-) activity by transfection with cDNAs for xCT and 4F2hc made A2780 cells more resistant to CDDP. Similar results on the cystine uptake were obtained in human colonic cancer cell lines. These findings suggest that the system x(c)(-) plays an important role in maintaining the higher levels of GSH and consequently in CDDP resistance in cancer cell lines. PMID- 12644837 TI - Role of platelet-derived endothelial cell growth factor/thymidine phosphorylase in fluoropyrimidine sensitivity. AB - Platelet-derived endothelial cell growth factor (PD-ECGF)/thymidine phosphorylase (TP) catalyses the reversible phosphorolysis of thymidine to thymine and 2 deoxyribose-1-phosphate and is involved in the metabolism of fluoropyrimidines. It can also activate 5'-deoxyfluorouridine (5'DFUR) and possibly 5-fluorouracil (5FU) and Ftorafur (Ft), but inactivates trifluorothymidine (TFT). We studied the contribution of TP activity to the sensitivity for these fluoropyrimidines by modulating its activity and/or expression level in colon and lung cancer cells using a specific inhibitor of TP (TPI) or by overproduction of TP via stable transfection of human TP. Expression was analysed using competitive template-RT PCR (CT-RT-PCR), Western blot and an activity assay. TP activity ranged from nondetectable to 70678 pmol h(-1) 10(-6) cells, in Colo320 and a TP overexpressing clone Colo320TP1, respectively. We found a good correlation between TP activity and mRNA expression (r=0.964, P&<0.01) in our cell panel. To determine the role of TP in the sensitivity to 5FU, 5'DFUR, Ft and TFT, cells were cultured with the various fluoropyrimidines with or without TPI and differences in IC(50)'s were established. TPI modified 5'DFUR, increasing the IC(50)'s 2.5- to 1396-fold in WiDR and Colo320TP1, respectively. 5-Fluorouracil could be modified by inhibiting TP but to a lesser extent than 5'DFUR: IC(50)'s increased 1.9- to 14.7-fold for WiDR and Colo320TP1, respectively. There was no effect on TFT or Ft. There appears to be a threshold level of TP activity to influence the 5'DFUR and 5FU sensitivity, which is higher for 5FU. Even high levels of TP overexpression only had a moderate effect on 5FU sensitivity. PMID- 12644838 TI - IDN 5390: an oral taxane candidate for protracted treatment schedules. AB - The recognition of the antiangiogenic properties of taxanes provides a basis for novel therapeutic approaches. A prolonged exposure to low drug concentrations has been proposed to be the most suitable approach to exploit the antiangiogenic potential of cytotoxic agents. Such schedule is required to target preferentially slowly dividing endothelial cells. The protracted use of taxanes could benefit from the availability of a taxane endowed with a favourable tolerability profile. Among compounds of a novel series of C-seco taxanes, IDN 5390 was originally selected on the basis of its potent antimotility activity and poor cytotoxicity on endothelial cells. The aim of the study was to investigate the preclinical pharmacologic profile of IDN 5390 in a variety of human tumour xenografts, including ovarian and colon carcinoma and a glioblastoma. IDN 5390, delivered by s.c. injection, daily for 5 days per week, exhibited a high activity against all tumours investigated (tumour growth inhibition was always >85%) in the range of well-tolerated doses. The maximum tolerated dose/injection (MTD), with no signs of systemic or local vesicant toxicity, was 120 mg kg(-1). In contrast, paclitaxel, delivered according to the same schedule, exhibited a variable antitumour efficacy associated with a substantial local toxicity (MTD=10 mg kg( 1)). Considering the remarkable efficacy of IDN 5390 delivered s.c. by protracted treatment schedule, the oral route of administration was further investigated, as the most suitable for daily treatment. Indeed, a good bioavailability of oral IDN 5390 was found. Oral IDN 5390 maintained a substantial efficacy against human tumour xenografts, including paclitaxel-resistant tumours, without loss of potency with respect to s.c. administration. In conclusion, the therapeutic advantages of IDN 5390, over paclitaxel, in protracted daily treatment schedules are represented by the oral efficacy and the high tolerability, which are favourable features to exploit the antiangiogenic potential and to design combinations with other effective agents. PMID- 12644839 TI - Reversal of Taxol resistance in hepatoma by cyclosporin A: involvement of the PI 3 kinase-AKT 1 pathway. AB - Hepatoma cells are known to be highly resistant to chemotherapy. Previously, we have found differential Taxol resistance in human and murine hepatoma cells. The aim of this study was to examine the effect of a multidrug resistance inhibitor, cyclosporin A in combination with Taxol on hepatoma in vitro and in vivo, and to identify the possible mechanism involved in Taxol resistance. Simultaneous treatment of cyclosporin A (0-10 microM) and Taxol (0.1 microM) inhibited cell growth in vitro. Cyclosporin A interfered with Taxol (0.1 microM)-induced AKT activation and BAD phosphorylation. Cyclosporin A combined with Taxol treatment augments caspase-9, -3 activation and loss of mitochondrial membrane potential in HepG2 cells. PI3 kinase inhibitor, wortmannin, or a dominant-negative AKT1 expression vector treatment partially enhanced Taxol-induced apoptosis indicating that PI3 kinase-AKT pathway was involved in Taxol-resistance pathway. Moreover, combination treatment reduced tumour growth in SCID and C57BL/6 mice as compared to either Taxol or cyclosporin A treatment. Our results indicate that the combination of cyclosporin A and Taxol is effective in the reversal of Taxol resistance through the inhibition of PI3 kinase-AKT1 pathway. PMID- 12644840 TI - Value of tyrosinase RNA detection by an RT-PCR method in melanoma prognosis. PMID- 12644842 TI - Correlation of cytochrome P450 (CYP) 1A2 activity using caffeine phenotyping and olanzapine disposition in healthy volunteers. AB - Olanzapine has previously been shown to have predominant metabolism by cytochrome (CYP) P450 1A2. Caffeine has been shown to provide an accurate phenotypic probe for measuring CYP1A2 activity. The purpose of this study is to determine if a significant correlation exists between olanzapine disposition and caffeine metabolic ratios. Subjects were phenotyped for CYP1A2 activity with caffeine probe methodology. After 200-mg caffeine administration, blood (4 h), saliva (6 and 10 h), and urine (8 h total) were collected for high-performance liquid chromatography (HPLC) analysis of caffeine and its metabolites.CYP1A2 activity was measured as plasma (PMR(4 h)), saliva (SMR(6 h) and SMR(10 h)), and three urinary metabolic (UMR1(8 h), UMR2(8 h), and UMR3(8 h)) ratios. Each of the 14 healthy nonsmokers (13 male) received a single 10 mg olanzapine dose after which blood was collected for HPLC determination of olanzapine concentrations at predose and at 0.5, 1, 1.5, 2, 2.5, 3, 4, 5, 6, 8, 10, 12, 24, 48, 72, 96, and 120 h postdose. Olanzapine pharmacokinetic parameters in this study were similar to those previously published. All caffeine metabolic ratios (PMR(4 h), SMR(6 h), SMR(10 h), UMR1(8 h), and UMR2(8 h)) significantly correlated with each other (p <0.001) except for UMR3(8 h), which did not correlate. A significant correlation (p <0.05) was also found between olanzapine clearance and PMR(4 h) (r=0.701), SMR(6 h) (r=0.644), SMR(10 h) (r=0.701), UMR1(8 h) (r=0.745), and UMR2(8 h) (r=0.710). A negative correlation was observed between olanzapine clearance and UMR3(8 h) (r=-0.029, p=NS). A significant correlation was found between olanzapine clearance and various caffeine metabolic ratios. Interpatient variability in CYP1A2 activity may explain the wide interpatient variability in olanzapine disposition. Compounds that modulate CYP1A2 activity may be expected to alter olanzapine pharmacokinetics accordingly. PMID- 12644843 TI - The alpha 2-adrenoceptor antagonist idazoxan reverses catalepsy induced by haloperidol in rats independent of striatal dopamine release: role of serotonergic mechanisms. AB - The alpha(2)-adrenoceptor antagonist idazoxan may improve motor symptoms in Parkinson's disease and experimental Parkinsonism. We studied the effect of idazoxan on haloperidol-induced catalepsy in rats, an animal model of the drug induced extrapyramidal side effects in man. Catalepsy was induced by a subcutaneous (s.c.) injection of haloperidol (1 mg/kg) and measured by the bar test for a maximum of 5 min. At 3 h after haloperidol, rats were given 0.16-5.0 mg/kg s.c. idazoxan, and descent latency was measured 1 h later. Idazoxan potently reversed haloperidol-induced catalepsy with an ED(50) of 0.25 mg/kg. This effect was mimicked by the selective alpha(2)-adrenoceptor antagonist RS 15385-197 (0.3 and 1 mg/kg orally). We assessed how dopaminergic mechanisms were involved in the anticataleptic effect of idazoxan by studying its effect on dopamine (DA) release in the striatum, with the microdialysis technique in conscious rats. Idazoxan (0.3 and 2.5 mg/kg) had no effect on extracellular DA and did not modify the rise of extracellular DA induced by haloperidol, indicating that changes of striatal DA release were not involved in the reversal of catalepsy. The anticataleptic effect of 2.5 mg/kg idazoxan (haloperidol+vehicle 288+/-8 s, haloperidol+idazoxan 47+/-22 s) was attenuated in rats given an intraventricular injection of 150 microg of the serotonin (5-HT) neurotoxin 5,7-dihydroxytryptamine (haloperidol+vehicle 275+/-25 s, haloperidol+idazoxan 137+/-28 s). The 5-HT(1A) receptor antagonist WAY100 635 (0.1 mg/kg s.c.) did not affect the anticataleptic effect of idazoxan. The results suggest that idazoxan reversed haloperidol-induced catalepsy by a mechanism involving blockade of alpha(2)-adrenoceptors and, at least in part, 5 HT neurons. PMID- 12644844 TI - Influence of reboxetine on salivary MHPG concentration and cognitive symptoms among patients with alcohol-related Korsakoff's syndrome. AB - This study is based on the hypothesis of a paraventricular cerebral noradrenaline deficit in alcoholic Korsakoff's syndrome. In a randomized open study the effects of a 4-week treatment with the selective noradrenaline reuptake inhibitor reboxetine on (1) the salivary concentration of the noradrenaline metabolite MHPG and (2) changes in cognitive performance measured by the Mini Mental Status Test were examined. The study group consisted of 105 patients diagnosed with alcohol related Korsakoff's syndrome (ICD-10: F10.6). Korsakoff's patients showed a reduced concentration of salivary MHPG compared to healthy controls; this reduction did not correlate with the results of the Mini Mental Status Test. An increase in salivary MHPG was found together with an improvement in the Mini Mental Status Test both in the verum group treated with reboxetine and in the control group upon completion of the 4-week study. However, a subgroup with a shorter duration of disease (<1 year) was found to profit significantly from reboxetine treatment, as shown by improvements in cognitive performance. PMID- 12644846 TI - Dissociable hormonal, cognitive, and mood responses to neuroendocrine challenge: evidence for receptor-specific serotonergic dysregulation in depressed mood. PMID- 12644848 TI - A simple capacitive cell for the measurement of liquids dielectric constant under transient thermal conditions. AB - A simple device for the measurement of the complex dielectric permittivity of liquids in various thermodynamic states has been developed. It uses a cylindrical aluminium capacitor of a type currently applied in tuning antenna circuits. The capacitor is filled with the liquid solution under study. A comparison of its capacity is made with that of the nitrogen filled capacitor tested under the same thermal conditions. This comparison allows the determination of the real and imaginary part of the solutions permittivity as a function of temperature (between 150 and 300 K) and frequency (between 100 Hz to 2 MHz). After validating the technique with pure glycerol and pure 1,2-propanediol, spectroscopic measurements have been undertaken on pure and diluted 1,2-propanediol in water. Due to the low heat capacity and the high thermal conductivity of the capacitor, cooling rates of 40 K/min have been achieved inside the solution, allowing measurements in the supercooled liquid and vitreous states. Results are presented and discussed in terms of relaxation and the physical states of the sample. By selecting the required thermal conditions, this device permits the observation of thermal transitions, such as ice crystallisation, and measurements to be conducted in the unstable supercooled liquid state. These measurements are necessary in the development of an effective electromagnetic warming device for vitrified cryoprotective solutions. PMID- 12644849 TI - Seed cryopreservation and longevity of two Salix hybrids. AB - The effects of desiccation and storage temperature on the viability and longevity of willow seeds was investigated using two hybrids, Salix rehderiana x (Salix x capreola) [cross 458] and Salix x sericans x Salix viminalis [cross 512]. Freshly harvested seed of both crosses survived silica gel drying down to c. 3 to 5% moisture content. Hybrid 458 seed stored in liquid nitrogen (-196C) for 3 d retained viability when equilibrated to < or = 45% RH (pre-storage), showed slightly reduced survival at 65% RH and exhibited no survival at > or =82% RH. The level of survival after 68 d for seeds pre-equilibrated to either c. 10 or 65% (5 or 10% moisture) and stored at four temperatures was -196C > -20C > 2C > 16C. At all temperatures, drier seed stored better than wetter seed. For hybrid 512, seed longevity at 20C > 40C > 60C, and a 10% fall in pre-storage seed RH resulted in a c. 2-fold increase in longevity at each storage temperature. The response of hybrid willow seeds to desiccation and cooling raises possibilities for the long-term seed conservation of Salix species by cryopreservation. PMID- 12644851 TI - Recovery of potato apices after several years of storage in liquid nitrogen. AB - A method for the systematic cryopreservation of potato apices was developed by the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ) and the Institute for Crop and Grassland Science of the Federal Agricultural Research Centre (FAL, Braunschweig). Designed specifically for routine use in genebanks, this method uses a very simple ultra-rapid freezing approach and was applied to a wide range of varieties within the Federal Centre for Breeding Research on Cultivated Plants (BAZ, Quedlinburg) Potato Collection. After several years of storage in liquid nitrogen, shoot tips from a random sample of 51 varieties were thawed and the survival and shoot regeneration percentages compared to those measured immediately after freezing. There were no major changes in either survival or recovery of frozen apices. Data presented are not the outcome of a systematic experiment but from that accumulated during our work from 1992 to 1999. PMID- 12644850 TI - The incidence of mitotic abnormalities in cryopreserved eight-cell early and compacted mouse embryos. AB - The effects of slow freezing-rapid thawing on viability and chromosomal complement of eight-cell early and eight-cell compacted mouse embryos were investigated. The abnormalities connected with damage to the mitotic apparatus, and with chromosome damage were investigated in cryopreserved embryos by cytological analysis. The embryos were preserved using 1M glycerol as cryoprotectant and a slow cooling regime to -30 degrees C before transfer to liquid nitrogen. The proportion of mitotic abnormalities in compacted embryos was significantly higher (13.1%) than in early embryos (5.9%) and unfrozen control embryos (3.9%) in these studies performed after thawing. This was reflected after cryopreservation by a reduced viability of the embryos as judged by culture to the hatching blastocyst stage--compacted 8-cell embryos (71.8+/-4.7%) versus controls (78.4+/-5.9%) and cryopreserved early-stage 8-cell embryos (86.1+/-4.0%) -p<0.05 in each case. However, aneuploidy rates were low in all groups in both fresh and cryopreserved embryos (around 3%). PMID- 12644853 TI - Ultrastructure of carp Cyprinus carpio permatozoa after cooling, dilution and freeze-thawing. AB - Dilution of the carp spermatozoa with a cryoprotective medium was shown to inflict serious injuries to the fine structures of some spermatozoa. The amount of the injured spermatozoa increases after cryopreservation. Examination of the freeze-fracture micrographs of the heads of the carp spermatozoa, showed that their protoplasmic faces were covered with the particles in high concentrations. These particles do not form clusters after cooling the spermatozoa down to +5 degrees C and dilution with a cryoprotective medium. For the first time the authors observed the particles to form geometrically shaped regular formations on the outer membranes of the carp spermatozoa[pf1]. On the basis of the results obtained the authors propose that high susceptibility of the carp spermatozoa to osmotic exposures is apparently due to some particular features of the fine structures of their membranes and to their expected probably low heterogeneity. PMID- 12644852 TI - Development of a cryopreservation protocol for the microcyclic rust-fungus Puccinia spegazzinii. AB - The rust fungus Puccinia spegazzinii (Basidiomycotina: Uredinales) has been identified as a potential classical biological control agent for the invasive weed Mikania micrantha (Asteraceae). Long-term, live storage of this pathogen is required for reference. As biotrophs, almost all rusts species cannot be preserved by traditional cryopreservation protocols, which rely on in vitro culture techniques. In addition, the embedded teliospores and delicate basidiospores of this microcyclic rust are not amenable to direct plunge freezing. Continuous culture of the rust on living plants is both laborious and expensive, so a variety of approaches for cryopreservation and storage were tested. These methods included traditional approaches to fungal cryopreservation such as variation of cooling rate regime and alginate encapsulation techniques. However, an in situ cryopreservation technique was the only method identified as having any potential for the long-term cryopreservation of the 10 isolates tested. Material from either petiole or stem tissue remained viable after cryopreservation, determined by the ability of the material to produce basidiospores. However, despite great progress being made in developing an optimal cryopreservation method, infection of the host plant by basidiospores produced from previously cryopreserved teliospores, embedded in leaf petioles, was not achieved. PMID- 12644854 TI - Cryopreservation of carrot (Daucus carota l.) cell suspensions and protoplasts by vitrification. AB - Carrot cell suspensions and protoplasts were successfully cryopreserved by vitrification. Cells were precultured in liquid Murashige and Skoog medium containing 0.175 M sucrose for 3 d and then in liquid MS medium containing 0.4 M sorbitol for 1 d. After loading of the precultured carrot cells in 25 % PVS2 at room temperature for 5 min and treatment with 100 % PVS2 at 0 degrees C for 7.5 min, they were quenched in liquid nitrogen. Optimal survival was 83.3 % (based on the triphenyl tetrazolium chloride reduction assay) following warming and unloading. Recovered cells retained the ability to regenerate plantlets in vitro. In the case of vitrification of protoplasts isolated from carrot cell suspensions, the optimal loading and dehydration durations were 5 min in 25% PVS2 and 3 min 100 % PVS2 respectively. Survival of 47 % of the untreated control (based on the FDA-PI (fluorescein diacetate-propidium iodide) staining) was achieved after cryopreservation. PMID- 12644855 TI - Skin hydration effects, film formation time, and physicochemical properties of a moisture mask containing Monostroma nitidium water-soluble mucilage. AB - The objectives of the study were to explore the effects of using the water soluble mucilage of Monostroma nitidium to replace the humectant and half of the thickening agent on the rheological properties, color, storage stability, water holding capacity, and film formation time of moisture masks thus prepared. Results showed that moisture masks containing water-soluble mucilage were pseudoplaxtic fluids. The apparent viscosity of these moisture masks decreased with increasing shear rate but increased with increasing concentration of the aqueous extracts used. The water-holding capacity of moisture masks containing 1% aqueous extracts and 1% hydroxyethyl cellulose (HEC) were similar to those containing 2% HEC and 5% 1-3 butadiene (humectant) but better than those containing 2% methyl cellulose (MC) and 5% humectant. The film formation time of moisture masks containing different concentrations of aqueous extracts decreased with increasing concentration of the aqueous extract used. The storage stability of a moisture mask containing 1% aqueous extract and 1% HEC was similar to that containing 2% HEC and 5% humectant and better than those containing 2% MC and 5% humectant. The safety test resulted in no erythema based on the Draize score test. The pH was between 7.1 and 7.5 for all moisture masks studied. PMID- 12644856 TI - Analysis of DNA in hair fibers. AB - The extraction and identification of deoxyribonucleic acid (DNA) from human hair shafts is described, along with the effects of hair treatments on levels of DNA and suggestions of DNA location within the shaft. DNA was present at low levels in the hair shaft, and was identified using polymerase chain reaction amplification of the human leukocyte antigen (HLA)-DQA1 locus. The use of cleanup columns aided the success of PCR amplification. DNA appears to reside in the cuticle portion of the hair shaft. Levels of DNA were found to be higher at the root-end compared to the tip-end of hair and were also found to be lower after permanent colorant treatment. DNA was found to be lost with surfactant washing, with increased loss occurring with prolonged or an increasing number of washes. These results suggest that small amounts of residual DNA remain after differentiation and add to our knowledge of the constituents of hair. PMID- 12644857 TI - Challenging the surfactant monomer skin penetration model: penetration of sodium dodecyl sulfate micelles into the epidermis. AB - The penetration of sodium dodecyl sulfate (SDS) into the epidermis was measured using (14)C-radiolabeled SDS. It was found that, at surfactant concentrations that exceed the critical micelle concentration (CMC) of SDS, the concentration of SDS measured in the epidermis increased as the total SDS concentration in the solution contacting the skin increased, thus demonstrating that micellar SDS contributes to the penetration of SDS into the epidermis. The observed SDS dose dependent response contradicts the widely accepted view that only surfactant monomers penetrate into the skin, while surfactant in micellar form does not contribute to surfactant penetration into the skin. Nevertheless, this finding is consistent with previously unexplained observations of a dose-dependent damage to the skin induced by SDS at concentrations above the CMC. When poly(ethylene oxide) (PEO) was mixed with SDS, SDS micelles bound to PEO did not contribute to the concentration of SDS in the epidermis, while SDS in free SDS micelles did. Dynamic light-scattering measurements revealed an average hydrodynamic radius of 20 A for the SDS micelles, and a larger radius of 25 A for the PEO-bound SDS micelles. A comparison with typical aqueous pore radii in the stratum corneum measured in the literature (10-28 A) suggests that the SDS micelles may be able to penetrate into the skin, while the PEO-bound SDS micelles may be sterically hindered from penetrating into the skin. PMID- 12644858 TI - Determination of methylparaben in o/w emulsions by solid-phase extraction and high-performance liquid chromatography. AB - A simple, fast, and accurate solid-phase extraction (SPE) using a 1-cc Oasis HLB cartridge for sample clean-up followed by an HPLC analysis for the assay of methylparaben (MP) in an o/w emulsion is described. One milliliter of methanol followed by 1 ml of 10% methanol in water was used to activate the cartridge sorbent. The sample was loaded into the cartridge and MP was then separated from oil-soluble excipients by washing the cartridge with 1 ml of 10% CH(3)CN in water. MP was finally eluted from the cartridge with mobile phase, acetonitrile and water (60:40), and quantified by HPLC analysis on a Nova-pak(R) C-18 column at 254 nm. PMID- 12644859 TI - Scalp hair length. I. Hair length in Florida theme parks: an approximation of hair length in the United States of America. AB - Ten hair length studies were conducted (January through May of 2001) at theme parks in central Florida, by observing and counting people with different lengths of hair with reference to specific anatomical sites. We observed 13.20% of the 24,300 males and females with hair shoulder-length or longer and 2.43% with hair down to the bottom of the shoulder blades or longer. We observed only 77 persons or 0.32% with hair waist-length or longer. Only three of these 77 persons were men. We observed 22 men with hair shoulder-length or longer or about 0.18%, among men only. Only four women, representing 0.0165% of this population, had hair reaching to the bottom of the buttocks. Hairstyles such as buns, topknots, or other styles of wearing hair bound to the head did interfere with our estimate of shorter lengths but not the longer ones (waist-length or longer). Head covers, e.g., caps, hats, scarves, etc., did not interfere with these estimates. PMID- 12644860 TI - The role of cationic conditioning compounds in reinforcement of the cuticula. PMID- 12644863 TI - Paediatric neurosurgery--a Southern African perspective. PMID- 12644864 TI - Endo-neuro-sonography: first clinical series (52 cases). AB - OBJECTIVE: A sono catheter for transendoscopic imaging was applied in neurosurgery for the first time in 52 patients with a broad variety of lesions. METHODS: A transendoscopic sono catheter (Aloka Deutschland GmbH, Dusseldorf, Germany) with a diameter of 1.9 mm (6F) was used and introduced into the working canal of an endoscope. The image produced by the probe is a 360 degrees scan ("brain radar") displayed on a monitor, on which some parameters can be varied to get the best view of the different anatomical structures. RESULTS: In 39 patients intraoperative imaging was the main reason for investigation and in 13 patients neuronavigation was the focus of interest. In 18 cases of tumor resection control targeting a visualized remnant was necessary. There are limitations and artifacts, which should reveal themselves in laboratory and clinical experience. CONCLUSION: In this small series, endo-neuro-sonography proved to make neuroendoscopy safer and easier by online and real-time imaging with high resolution. PMID- 12644865 TI - Long-term results of conventional treatment of diencephalic pilocytic astrocytoma in infants. AB - BACKGROUND: To develop an effective therapeutic strategy for patients with diencephalic pilocytic astrocytomas (PA), we must understand the long-term effects and problems of conventional treatments. METHODS: We performed a retrospective follow-up of four infants with PA in the diencephalon. CONCLUSION: The initial treatment consisted of partial tumor removal and/or external radiation. The mean progression-free duration after the initial treatment was 108.5 months. All patients received a second course of radiation to treat recurrent tumors. These conventional treatments produced relatively long survival. Three patients died 105, 202, and 379 months after the initial treatment. One patient is still alive at 249 months after the initial treatment. However, mental retardation was obvious in all four patients 6 to 12 years after the first irradiation and within 2 years of the second course of irradiation. Therefore, new therapeutic strategies must focus on delaying the timing of radiation until the patients are at least 5 years old and avoiding a second course of radiation to treat recurrence. PMID- 12644866 TI - Development-associated myristoylated alanine-rich C kinase substrate phosphorylation in rat brain. AB - OBJECT: In neuronal cells, myristoylated alanine-rich C kinase substrate (MARCKS), localized to particular areas of the synaptic membrane, is active during brain development. The destination of phosphorylated MARCKS is thought to be the cytoplasm where it is probably inactive. We compared MARCKS phosphorylation in the brains of embryonic, perinatal, and adult rats to determine its possible involvement in neurogenesis. METHODS: We prepared crude and partially purified extracts from various brain regions of rats aged between embryonic day 14 (E14) and 7 weeks after birth and assayed them for MARCKS phosphorylation by immunochemical methods. The isotypes of protein kinase C (PKC) were immunochemically identified in crude brain extracts from embryonic and postnatal rats. Despite negligible MARCKS phosphorylation, E16 brain extracts contained both MARCKS and PKCgamma, delta, epsilon, and lambda. MARCKS and polypeptides were clearly phosphorylated (49 and 45 kDa, respectively) in brain extracts purified on a DE52 column. Embryonic brain extracts manifested a high molecular-weight activity capable of suppressing polypeptide phosphorylation. This activity was markedly decreased on the day of birth and almost undetectable in the brains of 9-day-old rats. CONCLUSIONS: The embryonic rat brain appears to contain a protein(s) that suppresses the phosphorylation of other proteins including MARCKS. We posit that this inhibitory activity represents a factor(s) that plays a role in the regulation of neurogenesis beginning on the day on which MARCKS appears in the embryonic brain. PMID- 12644867 TI - Subdural hygroma: an unwanted result of Sylvian arachnoid cyst marsupialization. AB - INTRODUCTION: Subdural hygroma is a known complication of Sylvian fissure arachnoid cysts (SACs). However, most of the data in the literature refer to spontaneous or posttraumatic occurrence of subdural hygromas, regarded as either a favorable or an unfavorable event. Little is known about this phenomenon as a consequence of the surgical management of SACs. The present study was carried out to evaluate the significance of postoperative subdural hygromas in children with temporal arachnoid cysts, who have been treated with craniotomy and wide marsupialization of the cystic membrane. CLINICAL MATERIAL AND RESULTS: Between 1980 and March 2002, 104 children were operated on for a SAC at the Pediatric Neurosurgical Unit of the Catholic University Medical School in Rome. Six patients (5.8%; boys/girls=4/2; mean age 3.28 years) developed a subdural hygroma postoperatively, which required further treatment. According to Galassi's classification 4 children had a Type III cyst and 2 children a Type II cyst. All the patients had previously been submitted to open marsupialization of their arachnoid cyst and extensive removal of the cyst wall. In 5 patients (Type III cyst: 4 patients; Type II cyst: 1 patient), acute or subacute symptoms and signs of increased intracranial pressure (ICP) developed at a temporal distance, varying from 5 days to 1 month (mean interval: 14.3 days). All these children needed surgical treatment for the hygroma. A subduro-peritoneal (SDP) shunt was implanted as the first step in 3 patients (Type III cyst: 2 patients; Type II cyst: 1 patient). In the remaining 2 patients of this group (Type III cysts) medical therapy was initially attempted (acetazolamide: 24 mg/kg) without any improvement in clinical manifestations; on these grounds an external spinal subarachnoid drainage was implanted, but led only to the transient clearance of symptoms. The direct drainage of the subdural collection (SDP shunt in 1 patient and temporary external subdural drainage in the other), was followed by complete clinical recovery in both cases. The last patient in this series showed a pathologic increase in head circumference during the 1st month after surgery for his arachnoid cyst, followed by a subsequent stabilization. A CT scan documented a subdural hygroma, which first of all increased in size, but stabilized 2 months after surgery. No surgical treatment was performed in this case. At a mean follow up of 2.38 years (minimum: 1 month; maximum: 5 years) all the patients are in excellent clinical conditions. Complete neuroradiological resolution of the hygroma was observed in 2 of the 4 shunted patients, who have both been submitted to shunt removal (2 and 3 years after the implant respectively). CONCLUSIONS: It is our opinion that a wide opening of the external membrane of SACs may predispose the CSF fluid to accumulate within the subdural space, where its absorption is insufficient. A ball mechanism created by CSF pulsation may further increase the subdural fluid accumulation. Secondary distortion and occlusion at the level of the basal cisterns may also contribute to the persistence of the subdural fluid collection. Clinical manifestations may be transient, but frequently have a progressive course and the drainage of the subdural collection is, though transient, required in most cases. On these grounds we suggest the selective opening of the basal cisterns associated with a limited resection of the outer cyst walls in order to limit CSF access to the subdural space. PMID- 12644868 TI - The treatment of large supratentorial arachnoid cysts in infants with cyst peritoneal shunting and Hakim programmable valve. AB - OBJECTIVE: This retrospective case series examines 7 infants with large supratentorial arachnoid cysts who underwent cyst-peritoneal shunting and insertion of a Hakim programmable valve. Comparing pre- and postoperative clinical data, neuroradiological and regional cerebral blood flow (rCBF) findings we evaluated the efficacy of the surgical procedure. METHODS: Infants, ranging in age from 1 to 55 days (mean age 29.5 days), were assessed pre- and postoperatively by neurological examination, developmental profile and neuroimaging. RESULTS: Post procedure, all patients showed a significant reduction in the cyst/brain ratio on neuroimaging (p<0.001), 6 had a normal developmental profile (p<0.001) and 5 cases showed a significant amelioration of clinical symptoms and neurological signs. Two patients underwent preoperative SPECT scans, which showed hypoperfusion in the area surrounding the cyst; this decreased rCBF also improved post shunting. CONCLUSIONS: Large supratentorial arachnoid cysts in infants can be successfully treated with cyst-peritoneal shunting and insertion of a Hakim programmable valve. This is the first study specifically aimed at evaluating the long-term results of these conditions. PMID- 12644869 TI - Prognosis of brain stem lesion in children with head injury. AB - INTRODUCTION: Traumatic brain stem lesions (tBSL) in children are thought to be a rare phenomenon. This prospective study analyzed the frequency and significance of such lesions on early magnetic resonance imaging (MRI) after severe head injury, since CT fails to demonstrate these lesions. METHODS: In 30 consecutive children comatose after head injuries, MRI was performed within 8 days of the injury. RESULTS: The incidence of tBSL was 60%. When the lesion affected the pons or caudal portions of medulla oblongata bilaterally, mortality was 100%. The presence of tBSL significantly correlated with the duration of coma and the categories of outcome, as indicated by the Glasgow Outcome Score. The frequency and the distribution of tBSL in children were similar to adults. CONCLUSION: Magnetic resonance imaging appears to be of high predictive value after severe pediatric head injuries. PMID- 12644870 TI - Psychosocial functioning following surgical treatment for intractable epilepsy in childhood. AB - METHODS: Psychosocial functioning was examined in 13 children who had undergone surgical treatment for intractable epilepsy. The ratings of each child's social, emotional and behavioural problems were obtained via parent and teacher reports made an average of 2 years post-operatively. RESULTS: Parents viewed their children as having greater social difficulties than reported in standardisation samples. Although the overall level of difficulty fell below clinically significant cut-offs, the proportion of children who were rated as having clinically significant levels of difficulties approached significance. Teachers rated these children as displaying more symptoms of anxiety and depression, although the levels remained subclinical. DISCUSSION: These results are discussed in view of the complex interaction between neurological and environmental variables involved in social-emotional functioning following surgical treatment for epilepsy. PMID- 12644871 TI - Spinal cord involvement in pygopagus conjoined twins: case report and review of the literature. AB - INTRODUCTION: Pygopagus conjoined twins are joined at the rump and have union of the gastrointestinal and genitourinary systems as well occasional involvement of neural elements. CASE REPORT: We report a case where the terminal spinal cords were fused, despite which there was only a minor motor deficit in the lower limbs after separation. The pre-operative and intra-operative findings are described and issues of long-term follow-up discussed. DISCUSSION: Seven other cases reported in the literature with varying degrees of involvement of the spinal cord and cauda equina are reviewed. PMID- 12644872 TI - Intradural cerebellar chordoma in a child: a case report and review of the literature. AB - CASE REPORT: The authors describe an unusual case of an intradural cerebellar chordoma, the first such case to be reported in a child. A 9-year-old girl presented with headaches and papilloedema, and a discrete cerebellar mass was resected, which was shown histologically to be chordoma. There was no bony or dural association, and after radiologically proven complete resection it was elected that the child be followed expectantly. DISCUSSION: The case is compared with the few previously reported intradural chordomas, and insights into the origins and management of such cases are discussed. PMID- 12644873 TI - Cervical tuberculosis in early childhood. AB - CASE HISTORY: We present a case of spinal tuberculosis in a 4-year-old girl that grounds severe vertebral destruction involving four cervical vertebrae and a large abscess with retropharyngeal expansion. Presenting symptoms were neck and right arm pain, torticollis and weakness of the right arm. We drained the abscess and removed the infected bone and reconstructed the cervical spine with a fibular allograft. The neurological deficits were recovered and the torticollis was straightened up. CONCLUSION: We emphasize the value of early diagnosis of spinal tuberculosis and early radical surgery with spinal reconstruction. PMID- 12644874 TI - Pulsed Doppler tissue imaging and myocardial function in thalassemia major. AB - Few studies are available on left ventricular diastolic function in beta thalassemia major, and these have conflicting results. Our aim in this study was to analyze myocardial systolic and diastolic functions in patients with beta thalassemia major using, for the first time, pulsed Doppler tissue imaging (DTI), and compare data with standard Doppler echocardiography. We studied 30 young patients with beta-thalassemia major (age NTX) versus saline (1.0 ml/kg, IM, 4 h) followed by naltrexone (0.1 mg/kg, IM, 0.25 h) (SAL-->NTX). RESULTS: Seven subjects acquired the discrimination in an average of 108+/-14 sessions. MOR-->NTX-appropriate responding increased as an orderly function of increasing dose of morphine (0.56 1.7 mg/kg) and of naltrexone (0.01-10 mg/kg). The discrimination was also dependent upon interval between morphine and naltrexone administration. The MOR- >NTX cue was fully generalized to the combination of levorphanol (0.3 mg/kg) followed by naltrexone, but not to the non-opioid stereoisomer of levorphanol, dextrorphan (0.3 and 3.0 mg/kg) or the kappa-opioid-receptor-selective agonist U69,593 (0.3 mg/kg) followed by naltrexone. Naltrexone administered 15 min before morphine dose-dependently blocked MOR-->NTX-appropriate responding. CONCLUSIONS: This is the first non-rodent study of the discriminative effects of MOR-->NTX. MOR-->NTX produces a unique interoceptive stimulus that is pharmacologically selective, requires occupation of opioid receptors, presumably mu, for some minimum period of time, and is reversible. This discrimination procedure might provide new insights into the early drug-receptor interactions that underlie the development of physical dependence upon morphine-like drugs. PMID- 12644890 TI - Role of extracellular serotonin levels in the effect of 5-HT1B receptor blockade. AB - The release of serotonin (5-HT) at serotonergic nerve terminals is regulated by 5 HT(1B) autoreceptors. Several studies have reported that the effects of selective 5-HT reuptake inhibitors (SSRIs) on extracellular 5-HT are augmented by 5-HT(1B) receptor antagonists, whereas administration of these antagonists alone do not enhance 5-HT levels. It has been suggested that 5-HT(1B) receptors have low basal endogenous activity and therefore elevated endogenous 5-HT levels are needed to elicit an effect of 5-HT(1B) receptor antagonists. To test this hypothesis, different strategies were used to enhance 5-HT levels in the rat frontal cortex to assess the effects of locally applied NAS-181, a new selective 5-HT(1B) receptor antagonist. Blockade of 5-HT(1B) receptors with NAS-181 dose dependently augmented 5-HT levels when 5-HT levels were enhanced by a SSRI. No additional effect of NAS-181 on 5-HT output was found when 5-HT levels were enhanced by KCl depolarization-induced release or by preventing degradation of 5-HT with the monoamine oxidase inhibitor pargyline. In the presence of fluvoxamine, the increased 5-HT release evoked by KCl depolarization was augmented by NAS-181, supporting the idea that blockade of 5-HT transporters is necessary to measure an effect of 5-HT(1B) receptor blockade. In conclusion, the results provide circumstantial evidence that the effect of a 5-HT(1B) receptor antagonist depends on extracellular 5-HT levels, but strongly suggest that additional 5-HT reuptake inhibition is required to detect any effect of 5-HT(1B) receptor antagonist on 5 HT levels by in vivo microdialysis. PMID- 12644891 TI - Amphetamine increases schedule-induced drinking reduced by negative punishment procedures. AB - RATIONALE: d-Amphetamine has been reported to increase schedule-induced drinking punished by lick-dependent signalled delays in food delivery. This might reflect a drug-behaviour interaction dependent on the type of punisher, because no such effect has been found when drinking was reduced by lick-contingent electric shocks. However, the anti-punishment effect of amphetamine could be mediated by other behavioural processes, such as a loss of discriminative control or an increase in the value of delayed reinforcers. OBJECTIVES: To test the effects of d-amphetamine on the acquisition and maintenance of schedule-induced drinking reduced by unsignalled delays in food delivery. METHODS: Rats received 10-s unsignalled delays initiated by each lick after polydipsia was induced by a fixed time 30-s food reinforcement schedule or from the outset of the experiment. Yoked control rats received these same delays but independently of their own behaviour. d-Amphetamine (0.1-3.0 mg/kg) was then tested IP. RESULTS: d-Amphetamine dose dependently increased and then decreased punished schedule-induced drinking. The drug led to dose-dependent reductions when the delays were not contingent or when they were applied from the outset of training. CONCLUSIONS: These results support the contention that d-amphetamine has an increasing effect on schedule-induced drinking that has been previously reduced by a negative punishment procedure. This effect cannot be attributed to other potentially involved processes, and therefore support the idea that drug effects on punished behaviour depend on punishment being delays in food or shock deliveries. PMID- 12644892 TI - Genetic variation in the psychomotor stimulant properties of cocaine in Mus musculus. AB - RATIONALE: The psychomotor stimulant properties of drugs are argued to be a key feature of abuse liability. Several studies, primarily using inbred strains of mice, have demonstrated genetic variation in the psychomotor stimulant properties of cocaine. As of yet, however, no gene(s) has been identified which influences this phenotype. OBJECTIVES: The purpose of the present study was to examine a number of inbred strains of mice, including several closely related substrains, for cocaine-induced locomotor activation. Such substrain differences would suggest the possibility of a major gene effect. These data will also help to further characterize the range of genetic variation in response to cocaine. METHODS: Mice from 11 inbred strains were initially injected with saline and activity monitored for 30 min; mice were then removed from the activity monitor, injected with saline or one of six doses of cocaine, and activity was monitored for an additional 30 min. RESULTS: Compared to several other closely related C57BL substrains, we found the C57BL/10SnJ substrain to be significantly less activated following cocaine administration. In contrast, the C57BR/cdJ and C57L/J substrains showed extremely high levels of cocaine-induced locomotor activation. CONCLUSIONS: The genetic similarity between C57BL/10SnJ and the other closely related C57BL substrains suggests the possibility that the aberrant behavioral response to cocaine observed in B10SnJ mice may be due to a major gene effect. Similarly, the differences found in the C57BR/cdJ and C57L/J substrains may also be influenced by a major gene. The strains examined in this study will be useful tools for identification of relevant quantitative trait loci. PMID- 12644894 TI - Regulation of human insulin gene transcription by the immunosuppressive drugs cyclosporin A and tacrolimus at concentrations that inhibit calcineurin activity and involving the transcription factor CREB. AB - Cyclosporin A and tacrolimus are important immunosuppressive drugs. They share a diabetogenic action as one of their most serious adverse effects. In a single study, tacrolimus (100 nM) inhibited human insulin gene transcription in the beta cell line HIT. Using transfections of a human insulin-reporter gene into HIT cells, the present study shows that this inhibition is seen only at high concentrations of tacrolimus and is not caused by cyclosporin A. However, after stimulation by the major second messengers in the regulation of the insulin gene, cAMP and depolarization-induced calcium influx, both tacrolimus and cyclosporin A inhibited human insulin gene transcription in a concentration-dependent manner with IC(50) values of 1 nM and 30 nM, respectively. A further analysis offers a mechanism for this effect by revealing that the activation by cAMP and calcium of human insulin gene transcription is mediated by the transcription factor cAMP responsive element binding protein (CREB) whose activity is inhibited by the immunosuppressants. These data demonstrate for the first time that cAMP- and calcium-induced activity of the human insulin gene is mediated by CREB and blocked by both tacrolimus and cyclosporin A at concentrations that inhibit calcineurin phosphatase activity. Since also the immunosuppressive effects of cyclosporin A and tacrolimus are thought to be secondary to inhibition of calcineurin, the present study suggests that inhibition of human insulin gene transcription by the immunosuppressants is clinically important and may contribute to their diabetogenic effect. PMID- 12644893 TI - Ischemic and pharmacological preconditioning induces further delayed protection in transgenic mouse cardiac myocytes over-expressing adenosine A1 receptors (A1AR): role of A1AR, iNOS and K(ATP) channels. AB - In this study we examined the hypotheses that over-expression of the adenosine A(1) receptor (A(1)AR) in transgenic mouse cardiac myocytes (A(1)AR-tgm) induces cellular protection against subsequent sustained simulated ischemia (SI); that the cellular protection induced by over-expression of A(1)AR in A(1)AR-tgm is mediated through inducible nitric oxide synthase (iNOS) and K(ATP) channels. Sub lethal simulated ischemia (SSI) and the A(1)AR agonists chloro- N(6)-cyclopentyl adenosine (CCPA) or (2 S)- N(6)-[2-endo-norbornyl]adenosine (S-ENBA) induce further, delayed cytoprotection, additional to the existing protection in A(1)AR tgm. Cellular injury and cell viability was measured by the release of lactate dehydrogenase (LDH) or creatine kinase (CK) into the medium and the amount remaining in the cells. The cellular resistance acquired by cardiac myocytes due to the over-expression of A(1)AR was reflected by the reduced release of LDH (in units/liter) from 44.94+/-1.46 (wild-type mouse cardiac myocytes, wt) to 29.59+/ 2.83 (A(1)AR-tgm, P<0.001). Conversely, LDH release from A(1)AR-tgm increased to 42.53+/-2.23 ( P<0.01) on exposure to 5-hydroxydecanoate (a mitochondrial K(ATP) channel blocker), to 45.93+/-2.90 ( P<0.01) on exposure to S-methylthiourea (an iNOS inhibitor) and to 56.04+/-3.00 ( P<0.01) on exposure to glibenclamide (a K(ATP) channel blocker). Treatment of A(1)AR-tgm is with SSI and the A(1)AR agonists chloro- N(6)-cyclopentyl adenosine (CCPA) or (2 S)- N(6)-[2-endo norbornyl]adenosine (S-ENBA) decreased the release of LDH from 46.44+/-0.57 (A(1)AR-tgm) to 42.08+/-0.48 (A(1)AR-tgm plus SSI, P<0.05), 38.03+/-1.16 (A(1)AR tgm plus CCPA, P<0.001) and 32.77+/-0.58 (A(1)AR-tgm plus S-ENBA, P<0.001). Our data suggest that the A(1)AR has a cytoprotective effect against subsequent sustained SI in A(1)AR-tgm and that the cellular protection induced by over expression of A(1)AR in A(1)AR-tgm depends on iNOS and K(ATP) channels. Further, SSI and the A(1)AR agonists CCPA or S-ENBA induce further, delayed cytoprotection in A(1)AR-tgm. PMID- 12644895 TI - Anandamide metabolism by fatty acid amide hydrolase in intact C6 glioma cells. Increased sensitivity to inhibition by ibuprofen and flurbiprofen upon reduction of extra- but not intracellular pH. AB - The metabolism of anandamide by fatty acid amidohydrolase (FAAH) at different intra- and extracellular pH values has been investigated in intact C6 rat glioma cells. The cellular uptake of anandamide at 37 degrees C was found to decrease by 28% when the extracellular pH (pH(e)) was reduced from pH 7.4 to pH 6.2. In contrast, a selective decrease in intracellular pH (pH(i)), accomplished by acidifying the cells followed by incubation in sodium-free buffer at pH 7.4, did not affect the uptake. Anandamide uptake was inhibited by (R)-ibuprofen, with pI(50) values of 3.05+/-0.57, 3.66+/-0.23 and 3.94+/-0.88 at pH(e) values of 7.4, 6.8 and 6.2, respectively. In the presence of phenylmethylsulfonyl fluoride, however, (R)-ibuprofen failed to inhibit the uptake of anandamide. A reduction in pH(e) from 7.4 to 6.2 produced a 17% reduction in the FAAH-catalyzed metabolism of anandamide in the intact C6 cells. However, an increased sensitivity of FAAH activity to inhibition by (R)-ibuprofen as well as (R,S)-flurbiprofen and (S) flurbiprofen was seen at a lower pH(e). For (R)-ibuprofen, pI(50) values of 3.57+/-0.08, 4.04+/-0.05 and 4.59+/-0.04 were found at pH(e) values of 7.4, 6.8 and 6.2, respectively. For (R,S)- and (S)-flurbiprofen, the pI(50) values at pH(e) 7.4 were 4.02+/-0.05 and 4.13+/-0.18, respectively at a pH(e) of 7.4, and 4.81+/-0.11 and 4.84+/-0.10, respectively, at a pH(e) of 6.2. In contrast, intracellular acidification did not affect either the rate of anandamide metabolism or its inhibition by (R)-ibuprofen or (S)-flurbiprofen. It is concluded that a reduction of extracellular pH produces an enhanced accumulation of the acidic NSAIDs ibuprofen and flurbiprofen into C6 glioma cells and thereby an inhibition of anandamide metabolism. PMID- 12644897 TI - Bisindole alkaloids from Strychnos guianensis are effective antagonists of nicotinic acetylcholine receptors in cultured human TE671 cells. AB - Several mono- and bisindole quaternary alkaloids isolated from the stem bark of Strychnos guianensis have recently been shown to be effective blockers of neuromuscular transmission in mice. In this study, we used a human clonal cell line (TE671) expressing muscle-type nicotinic acetylcholine receptors. The agonist carbamylcholine activated a receptor-mediated (86)Rb(+) efflux and this activation was antagonized by the indole alkaloids, the most active being bisindole bisquaternary compounds. The most effective antagonist, guiachrysine, had an IC(50) around 0.43 microM in the presence of 0.5 mM carbamylcholine, compared to 0.16 microM for d-tubocurarine, the most potent curarizing alkaloid. Guiaflavine and 5',6'-dehydroguiaflavine were slightly less effective. Monoindole compounds were 10 to 100 times less potent than bisindole alkaloids. Kinetic analysis showed that the inhibition of the carbamylcholine-dependent (86)Rb(+) efflux by guiaflavine was of mixed competitive and uncompetitive type. The competitive component (K(I)=0.21 microM) is presumably due to binding at the acetylcholine site, while the uncompetitive component (K'(I)=0.92 microM) may be due to open channel block. PMID- 12644898 TI - Apoptosis induction in human cancer cells by sungucine from Strychnos icaja root. AB - Sungucine (SG) and isosungucine (ISG) are bisindole alkaloids characterized by a 5'-23 link between the two parts of the compounds, which are till now specific to Strychnos icaja. In this work, SG and ISG were submitted to the NCI's in vitro 60 human tumor cell line screen, where SG showed interesting selectivity (6X) against the tested leukemia cell lines. In HL60-treated cells, apoptosis was demonstrated by observation of apoptotic bodies formation, and phosphatidylserine exposition at cell surface. In HeLa-treated cells, the analysis of cellular cycle by flow cytometry showed G1 accumulation and a small sub-G1 peak that could be related to DNA fragmentation characteristic of apoptosis. The eventual role of p53 was analyzed using wild-type HCT-116 colon cancer cells. Nevertheless, p53 and Bax expression were not modified in SG-treated cells. The cleavage of PARP by caspase-3 protease proved that apoptosis was also induced in this line. These results demonstrate that SG induces apoptosis, but also necrosis, in human cancer cell lines. PMID- 12644896 TI - Mechanisms involved in the loss of excitatory post-stimulus responses by inflammation. AB - AIM: Electrical stimulation of colonic muscles elicits a response during the stimulation period, and a transient excitation after the stimulus. Post-stimulus or "rebound" excitation has been linked to pathways involving inhibitory neurotransmitters, prostaglandins and substance P but the mechanism is incompletely understood. Because rabbit colitis is characterized by a loss of inhibitory neurotransmission we hypothesized it might affect the rebound response. Therefore we characterized rebound responses in non-inflamed and inflamed tissue by comparing the effect of antagonists/blockers of putative (nitric oxide [NO], ATP, substance P, prostaglandins) and new (serotonin) neurotransmitters. METHODS: Strips from rabbits with colitis induced by 2,4,6 trinitrobenzenesulfonic acid (TNBS) were subjected to electrical field stimulation. Because rebound responses are more prominent under nonadrenergic noncholinergic (NANC) conditions, the effect of specific antagonists (N(omega) nitro-L-arginine methyl ester (L-NAME), indomethacin, SR140333, methiothepin) on the rebound response was compared under normal and NANC conditions. RESULTS: NANC conditions increased rebound responses in non-inflamed strips, but this effect was reduced or abolished in inflamed strips. Rebound responses were reduced by pretreatment with the NO-synthase inhibitor, L-NAME, under NANC conditions in non inflamed strips but not affected in inflamed tissue. In contrast, the P(2) purine receptor antagonist, suramin, did not affect rebound responses in inflamed and non-inflamed strips. The effect of the cyclo-oxygenase inhibitor (COX), indomethacin, on rebound responses was reversed from excitatory to inhibitory by inflammation. Under NANC conditions rebound contractions were also reduced by the neurokinin-1 (NK(1)) antagonist, SR140333, both in normal and inflamed strips. The most pronounced reduction in rebound responses in inflamed and non-inflamed strips under normal conditions was observed with the 5-hydroxytryptamin (1,2) (5 HT(1,2)) antagonist, methiothepin. CONCLUSION: Rebound responses are mainly non cholinergic and involve NO, substance P, serotonin and inhibitory prostaglandins. In inflamed tissue the nitrergic pathway is absent, excitatory prostaglandins prevail and the cholinergic and tachykinergic components are relatively more important. However there remains an important serotonergic contribution. Our data suggest that inflammation damages different neural pathways to a different extent and is most selective for nitrergic pathways. PMID- 12644899 TI - Enhanced adenosine A(2B) mediated coronary response in reserpinised rat heart. AB - In this study, we investigated the effect of noradrenaline depletion on contractile recovery in rat isolated heart following myocardial ischaemia. Groups tested included control tissues and hearts from reserpinised rats. Reserpine 1 mg/kg s.c. was injected into rats 18 to 24 h prior to experiments. Hearts underwent 15 min global normothermic ischaemia followed by 30 min reperfusion. Functional data (end diastolic pressure (EDP), heart rate (HR), left ventricular developed pressure (LVDP), dP/dt(max), dP/dt(min)) showed that contractile function following ischaemia-reperfusion is unaffected by reserpinisation. However, pre- and post-ischaemic coronary flow rates (CFR) were increased by 16 to 38% in hearts from reserpinised rats versus control hearts. Pre-ischaemic CFRs in control hearts (11.17+/-0.67 ml/in(-1) x g tissue(-1), n=9) were significantly lower then CFRs derived from reserpinised rat hearts (14.57+/-0.72 ml/min(-1)/g tissue(-1), n=10). Post-ischaemic reactive hyperaemia was evident in all groups. CFRs in reserpinised hearts remained elevated when compared to pre-ischaemic values through reperfusion (P<0.05). Reserpine treatment did not significantly alter pre- or post-ischaemic adenosine efflux. The A(2B) adenosine receptor antagonist alloxazine (10 microM) attenuated pre- and post-ischaemic CFRs in both control and reserpinised hearts (P<0.05) without altering the hyperaemic response while the A(2A) adenosine receptor antagonist 8-(3-chlorostyryl) caffeine (1 microM) did not alter CFRs in both groups. The A(3) adenosine receptor antagonist MRS1191 (0.1 microM) increased CFR in control and reserpinised hearts (P<0.05). Catecholamine depletion with reserpinisation enhances the responsiveness of the coronary resistance vessels to endogenous adenosine through activation of the A(2B) adenosine receptor. PMID- 12644900 TI - The alpha2-adrenoceptor antagonist atipamezole reduces the development and expression of d-amphetamine-induced behavioural sensitization. AB - The possible effect of atipamezole, a potent and specific alpha(2)-adrenoceptor antagonist, on the development and expression of d-amphetamine-induced behavioural sensitization was evaluated in mice. Male (C57Bl/6J) mice were given daily doses of d-amphetamine (2 mg/kg). In addition, groups of mice received injections of atipamezole (0.3 or 1 mg/kg) 20 min before d-amphetamine or vehicle administration. Idazoxan (1 mg/kg) was used in some experiments to extend the results to other alpha(2)-adrenoceptor antagonists. Challenge doses of d amphetamine were administered to the mice on days 7-9 to evaluate the effects of alpha(2)-adrenoceptor antagonists on the d-amphetamine sensitization, evidenced by increased locomotor activation. Mice treated repeatedly with d-amphetamine developed strong locomotor sensitization that was reduced by pretreatment with alpha(2)-adrenoceptor antagonists. Acute atipamezole at both doses attenuated the expression of d-amphetamine-induced sensitization. Atipamezole at 1 mg/kg alone had no effect on locomotor activity, but the lower dose (0.3 mg/kg) increased locomotor activity after repeated administration. These results indicate that alpha(2)-adrenoceptor antagonists modulate the actions of d-amphetamine in a manner not explicable by their enhancing actions on noradrenaline and dopamine release, and may thus provide a novel approach to the treatment of motor complications caused by dopaminergic agents, such as dyskinesias, and perhaps also drug dependence. PMID- 12644901 TI - Effects of the chromanol HMR 1556 on potassium currents in atrial myocytes. AB - PURPOSE: The chromanol HMR 1556 is a potent blocker of KvLQT1/minK potassium channels expressed in Xenopus oocytes. The compound is therefore a new class III antiarrhythmic drug with a distinct mechanism of action. However, the effect of HMR 1556 on atrial ion channels and the selectivity of block in the human heart has not been investigated. We tested the effects of HMR 1556 on repolarizing potassium currents in human and guinea pig atrial myocytes. METHODS AND RESULTS: Single atrial myocytes were isolated by enzymatic dissociation. Atrial potassium currents (I(Ks), I(Kr), in guinea pig, I(to), I(Kur), I(K1) in humans) were recorded at 36 degrees C in the whole cell mode of the patch clamp technique. HMR 1556 produced a concentration-dependent and reversible block of I(Ks) with a half maximal concentration (EC(50)) of 6.8 nmol/l. 10 micromol/l HMR 1556 almost completely inhibited I(Ks) (97.2+/-3.2%, n=6). Steady-state activation as well as kinetic properties of the current were not altered by HMR 1556. I(Kr) currents were not affected up to concentrations of 10 micromol/l. HMR 1556 did not inhibit other potassium currents in human atrium: I(to), I(Kur) and the classical inward rectifier potassium current I(K1) were not significantly affected up to concentrations that completely blocked I(Ks) (10 micromol/l). CONCLUSIONS: HMR 1556 is a highly-potent blocker of I(Ks) channels without exerting effects on other potassium currents involved in atrial repolarization. Given the potential advantages of I(Ks) vs. I(Kr) blockade, the drug's new mechanism of action warrants further investigation to clarify its role as an antiarrhythmic agent. PMID- 12644902 TI - Cell-specific toxicity of fibrates in human embryonal rhabdomyosarcoma cells. AB - The effects of a variety of fibrates on the cell viability were examined in human embryonal rhabdomyosarcoma cells (HRMSC). Five fibrates, including fenofibrate, clofibrate, gemfibrozil, bezafibrate and ciprofibrate, all concentration dependently reduced the cell viability determined by the mitochondrial enzyme activity. The cell injury occurred time-dependently and was marked at 24-48 h. The toxic action of fibrates was specific to HRMSC, since bezafibrate did not induce any marked changes in the viability of human microvascular endothelial cells or arterial smooth muscle cells. Synergistic cell injury was observed after a combined treatment with bezafibrate and simvastatin, although simvastatin alone reduced the cell viability. The cell injury was characterized by a typical nuclear damage, as evidenced by Hoechst 33342 staining and deoxynucleotidyl transferase dUTP nick-end label-positive staining. Similar cell-specific injury was induced by 8(S)-hydroxyeicosatetraenoic acid, a potent peroxisome proliferator-activated receptor alpha (PPARalpha) agonist. Consistent with these data, a marked expression for PPARalpha mRNA was observed in HRMSC but not in the endothelial or smooth muscle cells. Therefore, it is suggested that fibrates cause a cell-specific injury in HRMSC via activation of PPARalpha. Moreover, our present cell injury model using HRMSC may be useful for elucidating the mechanisms of clinical rhabdomyolysis induced by lipid-lowering agents. PMID- 12644903 TI - Effects of selective serotonin and serotonin/noradrenaline reuptake inhibitors on extracellular serotonin in rat diencephalon and frontal cortex. AB - Some clinical reports suggest that tricyclic antidepressants which block both noradrenaline and serotonin (5-HT) reuptake (SNRIs) are more effective than selective 5-HT reuptake inhibitors (SSRIs) in treating severe depression. Moreover, one neurochemical study reported larger increases in extracellular 5-HT in rat frontal cortex in response to the tricyclic antidepressant imipramine compared to the SSRI fluoxetine. However, imipramine, which blocks both 5-HT and noradrenaline reuptake, also binds with relatively high affinity to receptors for noradrenaline, histamine and acetylcholine. Thus, to test the hypothesis that compounds that inhibit both 5-HT and noradrenaline reuptake produce larger increases in 5-HT efflux, we compared the effects of acute systemic administration of several SNRIs and SSRIs. Extracellular 5-HT was measured using microdialysis probes implanted in the diencephalon and frontal cortex of unanesthetized rats. We tested the SSRIs paroxetine (0.3-10 mg/kg), citalopram (10-20 mg/kg) and fluoxetine (10 mg/kg), the nonselective tricyclic antidepressant imipramine (20 mg/kg) and the more selective SNRIs duloxetine (3 30 mg/kg) and venlafaxine (30-50 mg/kg). During the lights-off period, paroxetine and duloxetine increased 5-HT in the diencephalon approximately 300 and approximately 200%, respectively. During the lights-on period, paroxetine and duloxetine each increased 5-HT approximately 400% in the diencephalon. In the frontal cortex, both paroxetine and duloxetine increased 5-HT approximately 200%. Citalopram and venlafaxine each increased 5-HT in the diencephalon approximately 300%. Fluoxetine and imipramine increased 5-HT in the diencephalon by approximately 125 and approximately 80%, respectively. Thus, these results do not support the hypothesis that compared to SSRIs, compounds which inhibit both 5-HT and noradrenaline reuptake have a larger acute effect on extracellular 5-HT. PMID- 12644904 TI - Protective effects of calcitonin gene-related peptide-mediated evodiamine on guinea-pig cardiac anaphylaxis. AB - Previous investigations have indicated that the pharmacological effects of evodiamine, a major alkaloidal component of the dried, unripe fruit of Evodia rutaecarpa Bentham (Rutaceae), are associated with stimulation of calcitonin gene related peptide (CGRP) release and CGRP prevents cardiac anaphylactic injury. In the present study, the protective effects of evodiamine on cardiac anaphylaxis were examined. Presensitized guinea-pig hearts challenged with specific antigen (bovine serum albumin) caused a marked decrease in coronary flow, left ventricular pressure and its derivatives (+/-dp/dt(max)), an increase in heart rate, and prolongation of P-R interval. Evodiamine (0.3 microM or 1 microM) markedly increased the content of CGRP in the coronary effluent concomitantly with a significant improvement of cardiac function and alleviation of the extension of P-R interval. Evodiamine at the concentration of 1 microM also inhibited the sinus tachycardia. The protective effect of evodiamine on cardiac anaphylaxis was abolished by CGRP(8-37), the selective CGRP receptor antagonist. These results suggest that evodiamine possesses a protective effect of cardiac anaphylactic injury and that the effect of evodiamine is related to stimulation of CGRP release. PMID- 12644905 TI - Angiotensin II subtype AT1 receptor blockade prevents hypertension and renal insufficiency induced by chronic NO-synthase inhibition in rats. AB - Aim of the present study was to investigate the influence of the angiotensin II (ANG II) subtype 1 (AT(1)) receptor blockers fonsartan and losartan on blood pressure, cardiac -dynamics and -metabolism as well as functional and morphological changes in the kidney of rats after long-term inhibition of the nitric oxide (NO) synthase by N(G)-nitro-L-arginine methyl ester (L-NAME). Oral chronic treatment with L-NAME in a dose of 25 mg/kg/d over 6 weeks caused a significant increase in systolic blood pressure (198+/-13 mmHg) when compared to untreated rats (144+/-4 mmHg). Animals receiving simultaneously L-NAME and fonsartan (10 mg/kg/d) or losartan (30 mg/kg/d) were protected against blood pressure increase. L-NAME treatment caused a significant decrease in glomerular filtration rate (GFR) from 4.52+/-0.81 to 1.34+/-0.26 ml/kg(-1)/min(-1) and renal plasma flow (RPF) from 10.52+/-1.29 ml/kg(-1)/min(-1) to 5.66+/-1.06 ml/kg( 1)/min(-1). Co-treatment with fonsartan and losartan prevented L-NAME-induced reduction in GFR and RPF. There was no difference in urine, sodium and potassium excretion in groups under investigation. Plasma renin activity (PRA) was further stimulated by fonsartan and losartan treatment. L-NAME produced a significant elevation in urinary protein excretion which was antagonised by both AT(1) blockers. Isolated hearts from animals treated with L-NAME showed a significant prolongation in the duration of ventricular fibrillation and a significant decrease in coronary flow as compared to control hearts. Treatment with fonsartan and losartan significantly decreased the duration of ventricular fibrillation as compared to L-NAME group. In addition, both AT(1) blockers given alone significantly reduced the duration of ventricular fibrillation as compared to hearts from untreated controls. During ischemia the cytosolic enzymes lactate dehydrogenase and creatine kinase as well as lactate in the coronary effluent were significantly increased in the L-NAME group. Myocardial tissue values of glycogen, ATP, and creatine phosphate were decreased, whereas lactate was increased. Fonsartan and losartan treatment totally abolished these effects. Histological examination of kidneys revealed that simultaneous administration of fonsartan and losartan with L-NAME abolished L-NAME-induced arteriolar hyalinosis, segmental sclerosis of glomerular capillaries and focal tubular atrophies. In conclusion, long-term blockade of ANG II subtype AT(1) receptors by fonsartan and losartan prevented L-NAME-induced hypertension, renal insufficiency, as well as cardio-dynamic, cardio-metabolic, and morphological deteriorations. PMID- 12644906 TI - Inhibition of skeletal muscle sodium currents by mexiletine analogues: specific hydrophobic interactions rather than lipophilia per se account for drug therapeutic profile. AB - In striated fibers, the activity of mexiletine (Mex)-like sodium channel blockers is strongly modulated by the part of the molecule nearby the asymmetric carbon atom. A lipophilic aromatic phenyl group at this levels, as in 2-(2,6 dimethylphenoxy)-1-phenylethanamine (Me4), markedly increases drug potency, while an increased distance between the stereogenic center and the pharmacophore amino group, as in 3-(2,6-dimethylphenoxy)-2-methylpropan-1-amine (Me2), enhances the use-dependent behavior. In order to better evaluate the role of lipophilicity in drug potency in relation to the structural determinants for a specific binding, lipophilic analogs of Me2 and Me4 were synthesized. Compounds 3-[(2,6 dimethylphenyl)thio]-2-methylpropan-1-amine and 2-[(2,6-dimethylphenyl)thio]-1 phenylethanamine were obtained by isosteric substitution of the oxygen atom with sulfur, while the introduction of a chlorine atom in 4- position of the aryloxy ring lead to 3-(4-chloro-2,6-dimethylphenoxy)-2-methylpropan-1-amine and 2-(4 chloro-2,6-dimethylphenoxy)-1-phenylethanamine. The compounds were tested on nearly maximal Na(+) currents elicited with depolarizing steps at 0.3 Hz (tonic block) and 2-10 Hz (use-dependent block) by means of vaseline-gap voltage-clamp method on single frog muscle fibers.The augmented lipophilicity largely increase drug potency in Me2 analogues, the thio and chlorinated compounds being 20- and 10-fold more potent in producing the tonic block, respectively. However, both compounds showed a 2-fold lower use-dependent behavior vs. the high use-dependent Me2. Surprisingly, the same increase in lipophilicity brought about by the same substitutions, in the already high lipophilic and potent Me4 failed to further improve the potency, although both new analogs were more stereoselective than Me4. No correlation was found between logP and potency of all analogs tested. All compounds acted as inactivated channel blockers. In conclusion, lipophilicity differently influences drug profile based on the molecular determinants controlling drug-receptor interaction. PMID- 12644907 TI - Molecular structure of the rabbit alpha2A-adrenoceptor: a contribution to the alpha2A-adrenoceptor versus I1 imidazoline receptor controversy. AB - In view of the high structural and pharmacological similarities between the alpha(2A)-adrenoceptors of humans and other mammalian species, it has been concluded, in particular, from experiments in rabbits that the (2A)-adrenoceptor is the exclusive site of action of central antihypertensive drugs, although the amino acid sequence of the alpha(2A)-adrenoceptor of just this species was unknown. Therefore, the aim of the present investigation was to determine the complete nucleotide sequence of the coding region of the rabbit alpha(2A) adrenoceptor gene. Degenerate oligonucleotides corresponding to regions of the alpha(2A)-adrenoceptor conserved between rat and man were used in a polymerase chain reaction with genomic DNA prepared from rabbit. A 1,356-base pair product with an open reading frame of 1,353 base pairs was obtained that encodes a protein of 451 amino acids which is similar to the alpha(2A)-adrenoceptors of other mammals (man, pig, rat, mouse, guinea-pig and cattle) but not to their alpha(2B)- and alpha(2C)-adrenoceptor subtypes suggesting its classification as an alpha(2A)-adrenoceptor. However, the degree of amino acid sequence identity is, at best, only 80% and, thus, about 10% less than between the other mammalian species. Compared with the human sequence there are 81 substantial changes of amino acids. In conclusion, rabbit and human alpha(2A)-adrenoceptors substantially differ in their amino acid sequence which may explain the opposite pharmacodynamic properties of the central antihypertensive drug rilmenidine (alpha(2)-adrenoceptor agonism and antagonism, respectively) reported in the literature. Hence, the present study supports the view that experiments with central antihypertensive drugs in rabbits are not reliably predictive for the site of action of such drugs in man. PMID- 12644908 TI - Is autopsy dead in the ICU? PMID- 12644909 TI - The perplexing role of copper-zinc superoxide dismutase in amyotrophic lateral sclerosis (Lou Gehrig's disease). AB - The existence of a link between some cases of familial amyotrophic lateral sclerosis (FALS) and copper-zinc superoxide dismutase (CuZnSOD) has been understood for almost a decade. However, beyond the fact that mutations in CuZnSOD cause FALS by a toxic gain of function, the mechanism whereby specific mutations in the protein structure result in development of the disease has remained almost a complete mystery to date. We have undertaken a critical survey of in vitro characteristics of over 30 of the 90 different CuZnSOD mutant proteins that are known to cause FALS in order to determine the differences that exist between mutant and wild-type properties. As-isolated metal content analysis, SOD activity assays, and thermal stability determinations of a significant fraction of the mutants show that the FALS mutant SOD proteins can be classified distinctly into one of two groups. Members of the first group, termed wild-type-like, have physical properties and enzymatic activities that are strikingly similar to those of wild-type CuZnSOD. The second group, however, show aberrant metal content in the as-isolated forms, compromised SOD activities, and unusual DSC thermoscans. All mutations in the members of this second group occur in or near the metal binding sites of the protein and thus they are termed metal binding region mutants. We have also compared the relative rates of self inactivation caused by reaction of the wild-type protein and several FALS-linked CuZnSOD mutants with hydrogen peroxide, as a measure of relative peroxidative activities. Results and implications of the role of CuZnSOD in FALS are discussed. PMID- 12644910 TI - Modulation of the NO trans effect in heme proteins: implications for the activation of soluble guanylate cyclase. AB - The binding of NO to the iron heme in guanylate cyclase and other heme proteins induces the cleavage of the proximal histidine bonded to the metal. In this study we assess by means of density functional theory (DFT) electronic structure calculations the role of H-bonding to histidine in the modulation of this effect. We have considered in the first place a model of the isolated active site coordinated with imidazole and imidazolate to mimic the effects of a very strong H-bond. We have also investigated four selected ferrous heme proteins with different proximal histidine environments: the O(2) sensing FixL, horseradish peroxidase C, and the alpha and beta subunits of human hemoglobin. Our results indicate that polarization and charge transfer effects associated with H-bonding to the proximal histidine play a fundamental role in the modulation of the NO trans effect in heme proteins. We also find computational evidence suggesting that protein structural constraints may affect significantly the cleavage of the Fe-His bond. PMID- 12644911 TI - New aspects of the pathogenesis of cystinosis. AB - Cystinosis is a lysosomal transport disorder characterized by an intra-lysosomal accumulation of cystine, the disulfide of the amino acid cysteine. It is the most common inherited cause of the renal Fanconi syndrome. There are various clinical forms, infantile, juvenile, and ocular, based on age of onset and severity of symptoms. The first clinical description appeared in the early 1900s, but it was not until 1998 that the causative gene, CTNS, was identified. CTNS encodes cystinosin, a novel seven transmembrane domain (TM) protein. Cystinosin is a lysosomal membrane protein that requires two lysosomal targeting signals: a classic GYDQL motif in its C-terminal tail and a novel conformational motif, the core of which is YFPQA, situated in the fifth inter-TM loop. Cystinosin is the lysosomal cystine transporter and its activity is H(+)-driven. A mouse model of cystinosis was recently generated and Ctns(-/-) mice accumulate cystine in all tissues. A high level of cystine accumulates in the kidney, but these mice do not present with proximal tubulopathy or renal dysfunction. The Ctns(-/-) mouse model may provide clues to the cause of the Fanconi syndrome associated with cystinosis, the origin of which remains poorly understood. PMID- 12644913 TI - Homocysteine, folate, vitamin B12 levels, and C677T MTHFR mutation in children with renal failure. AB - Hyperhomocysteinemia is well documented in chronic renal failure (CRF) and premature and progressive occlusive vascular disease is common in CRF. The combined effects of renal failure, folate and vitamin B(12) levels, and a common mutation (C677T) in the methylenetetrahydrofolate reductase (MTHFR) gene that leads to total plasma homocysteine (tHcy) elevation in CRF children were investigated. Forty-two children (15 females) with CRF, mean age 10.3+/-4.7 years, were included. The mean glomerular filtration rate (GFR) was 37.3+/-16.9 ml/min per 1.73 m(2). The control group comprised 33 children (18 females) with a mean age of 8.6+/-3.4 years. There were 40% of CRF patients with hyperhomocysteinemia. Folate and vitamin B(12) deficiencies were identified in 14% (n=6) and 5% (n=2), respectively, of all patients. On univariate analysis, the tHcy serum concentration was negatively correlated with the plasma folate concentration (P<0.05) in controls, and with GFR (P<0.05) in patients. On multiple regression analysis for the predictors of tHcy serum concentrations, folic and vitamin B(12 )were significant in controls, whereas only GFR was significant in CRF children. In our patients no effect of the MTHFR polymorphism on tHcy levels was seen This result, in addition to the limited number of patients, may partially be explained by the low prevalence of folate deficiency in our patients. PMID- 12644914 TI - Apolipoprotein E polymorphism and clinical course in childhood nephrotic syndrome. AB - Numerous studies have reported on the role of apolipoprotein E (apoE) polymorphism in the progression of diseases associated with lipid abnormalities. However, few studies have been performed to date on the relationship between apoE polymorphism and childhood nephrotic syndrome (NS). In the present study, we evaluated the allelic and genotypic frequencies of the apoE gene and the possible association between the polymorphic forms of the apoE gene on the clinical course in 190 patients with childhood NS, who were further classified into frequent relapsers (FR, 92) and nonrelapsers or infrequent relapsers (IR, 98). Controls included 132 healthy Koreans. Allele-specific primers were used to detect polymorphism of the apoE gene. The allelic frequencies at the apoE locus were 5.9%, 82.6%, and 11.8% for alleles epsilon2, epsilon3, and epsilon4, respectively in the childhood NS group, while those in the control group were 6.8% for epsilon2, 88.3% for epsilon3, and 4.9% for epsilon4. The allelic frequency for epsilon4 in childhood NS was twice that of controls. Moreover, the allelic frequency of the epsilon4 allele in the FR group was 3.4 times that of the control group and 2.5 times that of the IR group. The high frequency of epsilon4 in patients with childhood NS suggests that epsilon4 may serve as a genetic marker for predisposition to childhood NS. We believe that the apoE allele type is of considerable significance in predicting the course of the disease. PMID- 12644912 TI - Expression of connexins in the normal and obstructed developing kidney. AB - Connections between cells are achieved by proteins called connexins that comprise the gap junction. Connexins play a major role in organ development. Our reverse transcription-polymerase chain reaction (RT-PCR) studies demonstrate that Cx30, Cx36, Cx37, Cx40, Cx45, Cx46, and Cx50 are expressed in the kidney. Quantitative RT-PCR indicates that Cx37, Cx45, and Cx46 are preferentially expressed during early renal development. We also explored the expression of connexins in neonatal unilateral ureteral obstruction (UUO). After 12 days of neonatal UUO, the renal mRNA expression of Cx30, Cx37, and Cx40 was significantly elevated. In contrast, there was no change in connexin renal mRNA levels in adult UUO. We conclude that multiple connexins are expressed in the rat kidney and several are aberrantly expressed in neonatal UUO. PMID- 12644915 TI - Glomerular structural factors in progression of congenital nephrotic syndrome. AB - The glomerular structural factors associated with progression of congenital nephrotic syndrome of Finnish type (CNF) are incompletely understood. We studied glomerular volume and the proportion of various glomerular subtypes in seven CNF patients (aged 4 months to 3 years). Glomerular tubular connections (GTC) were analyzed in two patients early and late in the disease spectrum (creatinine 0.2 and 2.8 mg/dl, respectively). The proportions of "normal" glomeruli decreased (r= 0.91, P=0.003) and microcystic glomeruli increased (r=0.85, P=0.015), while the fetal and sclerosed glomeruli remained stable (r=0.073 and 0.08 respectively, P=NS for both) with age. The "normal" glomeruli were larger in CNF than in six age-matched minimal change nephrotic patients (P=0.009). GTC analyses in the mild disease showed that 75% of the 12 "normal" glomeruli had normal tubules, and 25% were atubular or were attached to atrophic tubules, while 88% of the 12 fetal glomeruli were either atubular or were attached to atrophic tubules. In the advanced disease, 67% of 12 "normal" glomeruli had normal tubules, and 33% were atubular or atrophic, while none of the fetal or microcystic glomeruli had "normal tubules". Thus atubular glomeruli may play an important role in disease progression in CNF, as it is associated with a progressive decrease in hypertrophied "normal" glomeruli and an increase in largely atubular microcystic glomeruli. PMID- 12644916 TI - Reflux nephropathy and hypertension: correlation with the progression of renal damage. AB - The aim of this study was to evaluate the relationship between blood pressure (BP), measured with ambulatory blood pressure monitoring (ABPM), and the progression of renal damage in 100 (70 females, 30 males) normotensive children with reflux nephropathy (RN). The patients, mean age of 13.5+/-5 years and almost 5 years of follow-up, were divided according to degree of RN into group A (I/II) and group B (III/IV). For each subject, 24-h systolic and diastolic BP (SBP-DBP), load (percentage of BP readings that exceeded the age- and sex-specific 95th percentile), and biochemical parameters were recorded. There was no significant difference in casual BP between the groups. The mean 24-h SBP-DBP and load were significantly higher in group B than A. There was a significant difference in creatinine (Cr) levels between the groups, and Cr correlated with BP in both groups. In group B, microalbuminuria correlated with ambulatory BP, and plasma renin activity failed to decrease with chronological age. Elevated load was shown in 8 of 50 patients in group A and in 21 of 50 in group B. In 3 of 12 patients of group B, with increased load BP, left ventricular geometry, by integrated backscatter, was abnormal. ABPM was useful in selected children at risk of hypertension. PMID- 12644917 TI - Shigatoxin-1 binding and receptor expression in human kidneys do not change with age. AB - Postdiarrheal hemolytic-uremic syndrome (D+HUS) occurs predominantly in young children. The rarity of D+HUS in adults has been ascribed to aging-associated loss of glomerular globotriaosylceramide (Gb3) expression, the major cognate receptor for shigatoxin. This belief, however, is based on relatively little data. The current study was undertaken to examine renal shigatoxin-1 (Stx-1) binding and Gb3 expression by human kidneys from varying aged subjects. Immunofluorescent staining and thin layer chromatography of neutral lipid extracts were performed. Abundant Stx-1 binding to both glomeruli and tubules was observed in frozen renal sections from all subjects of all ages (6 months to 85 years). The pattern of Stx-1 binding was identical between adults and children, with glomerular endothelial cells and cortical tubules being strongly labeled. Stx-1 binding affinity was similar between pediatric and adult kidneys. Antibodies to Gb3 showed a similar pattern and degree of staining regardless of donor age. In addition, Gb3 levels in glomeruli and tubules isolated from fresh kidney tissue were comparable between different aged donors. These data demonstrate that intrinsic renal binding of Stx-1 does not vary with age. It is suggested that factors other than basal renal Gb3 expression account for the age related incidence of acute renal failure in D+HUS. PMID- 12644918 TI - Effect of iron deficiency anemia on renal tubular function in childhood. AB - Little is known about renal function in children with iron deficiency anemia. The purpose of this study was to investigate renal tubular function in these children. We compared renal tubular function in 20 children with iron deficiency anemia with 20 healthy age-matched controls. Blood and urine samples were obtained for hematological and biochemical analysis. Mean fractional excretion of sodium and mean urinary N-acetyl-beta- D-glucosaminidase/creatinine were significantly higher in the children with iron deficiency anemia than in controls (P<0.05). Hemoglobin levels were negatively correlated with urinary N-acetyl-beta D-glucosaminidase/creatinine (r= -0.44, P=0.015), but were not correlated with fractional excretion of sodium (r= -0.29, P=0.13). There was no correlation between urinary N-acetyl-beta- D-glucosaminidase/creatinine and fractional excretion of sodium (r=0.32, P=0.09). The results suggest that children with iron deficiency anemia have impaired renal tubular function. PMID- 12644919 TI - Renal tubular dysfunction in alpha-thalassemia. AB - Shortened red cell life span and excess iron cause functional and physiological abnormalities in various organ systems in thalassemia patients. In an earlier study, we showed that beta-thalassemia patients have a high prevalence of renal tubular abnormalities. The severity correlated with the degree of anemia, being least severe in patients on hypertransfusion and iron chelation therapy, suggesting that the damage might be caused by the anemia and increased oxidation induced by excess iron deposits. This study was designed to define the renal abnormalities associated with alpha-thalassemia and to correlate the renal findings with clinical parameters. Thirty-four pediatric patients (mean age 8.2+/ 2.8 years) with Hb H disease or Hb H/Hb CS were studied. Ten patients (group 1) were splenectomized, with a mean duration post splenectomy of 3.5+/-1.4 years; 24 patients (group 2) had intact spleens. The results were compared with 15 normal children. Significantly higher levels of urine N-acetyl-beta- d-glycosaminidase, malondialdehyde (MDA), and beta(2)-microglobulin were found in both groups compared with normal children. An elevated urine protein/creatinine ratio was recorded in 60% of group 1 and 29% of group 2. Two patients (5.9%), 1 in each group, had generalized aminoaciduria. We found proximal tubular abnormalities in alpha-thalassemia patients. Increased oxidative stress, possibly iron induced, may play an important role, since urine MDA levels were significantly increased in both groups of patients. PMID- 12644920 TI - Urea percentiles in children with chronic renal failure. Data from the ItalKid project. AB - In chronic renal failure high serum urea levels (sUrea) are correlated with the onset of uremic symptoms. Urea has generally been considered relatively non toxic, functioning more as a surrogate for other toxic solutes; however, it has been recently reported that it can contribute to uremic toxicity. Clinically sUrea are often difficult to interpret because of the wide range of kidney functions. To obtain a practical and easily accessible tool to evaluate sUrea, we have produced percentile curves for different ranges of chronic renal failure, defined with creatinine clearance ( C(Cr)) obtained with the Schwartz formula. Data were obtained from the Italian Pediatric Registry of Chronic Renal Failure (ItalKid); its inclusion criteria are: (1) C(Cr )<75 ml/min per 1.73 m(2), (2) age <20 years at time of registration, and (3) conservative treatment. To obtain the percentiles, the following patients were excluded: patients with an underlying disease, a concomitant treatment, or a disorder that could affect urea metabolism, per se, and/or food intake, and patients aged <2 years. The study group included 690 subjects (mean age 9.56+/-4.54 years, 485 males). In total, 2,085 observations (C(Cr )and sUrea) were available for the construction of the percentile curves. A median of 258 (range 99-380) observations was obtained for each of the eight different categories of C(Cr )(intervals of 10 ml/min per 1.73 m(2)). The 10th, 25th, 50th, 75th, and 90th percentiles were calculated and a graph was produced. Patients with the highest urea percentiles showed significantly higher plasma levels of phosphorus and parathyroid hormone and significantly lower hemoglobin concentrations and bicarbonate levels. Our percentile curves may help to identify subjects with inappropriate sUrea for a given C(Cr). PMID- 12644921 TI - Mycophenolic acid pharmacokinetics in stable pediatric renal transplantation. AB - Mycophenolate mofetil (MMF) is given to children in fixed doses based either on body weight or body surface area. There are data indicating mycophenolic acid (MPA) blood levels should be monitored in the early period of transplantation. However, there is little information regarding MPA pharmacokinetics (PK) in stable pediatric recipients. We evaluated MPA-PK in 20 stable renal transplant children (11.7+/-1.9 years) under long-term (46+/-31 months) MMF (26.1+/-7 mg/kg per day or 785+/-183 mg/m(2) per day) therapy plus prednisone and cyclosporin A (n=16), tacrolimus (n=3), or MMF/prednisone (n=1). Total MPA levels were measured using the EMIT-MPA assay at 0, 1, 2, 3, 4, 6, and 8 h after an oral dose of MMF. The level at 12 h was considered equal to the trough level for AUC(0-12) calculation. Mean C(0), C(max), AUC (0-12), and T(max )were 3.46+/-1.32, 13.5+/ 0.58 microg/ml, 63.2+/-24.4 microg x h/ml, and 1.3+/-0.6 h, respectively. Six (30%) children were considered to have an adequate exposure (36-54 microg x h/ml) to MPA, 11 (55%) showed an AUC(0-12 )>54 microg.h/ml, and 3 (15%) showed an AUC(0 12 )<36 microg x h/ml. A C(max )>/=10 microg/ml was seen in 13 (65%) children. MMF dose did not correlate with AUC(0-12) or C(max). The combination of variables C(0), C(1), and C(4 )provided an equation to predict exposure (r(2)=0.75) where AUC(0-12)=12.62+(7.78 x C(0))+(0.90 x C(1))+(1.30 x C(2)) (P<0.001). The use of MMF without monitoring MPA blood levels may cause unnecessary overexposure to the drug in stable pediatric recipients. PMID- 12644922 TI - Mutated podocin manifesting as CMV-associated congenital nephrotic syndrome. AB - We report a girl with congenital nephrotic syndrome (CNS) associated with cytomegalovirus (CMV) infection and histological findings on renal biopsy that suggested a causal relationship between the two. She was subsequently found to be homozygous for a nonsense mutation in the NPHS2 gene encoding podocin (R138X), which is the true cause of her NS. Based on review of the literature and our findings in this patient, we propose that the clinical entity known as CMV causing CNS may not exist. PMID- 12644923 TI - Renal artery stenosis and nephrotic syndrome: a rare combination in an infant. AB - We describe an uncommon pediatric finding of unilateral renal artery stenosis, which presented as nephrotic syndrome, hypertension, failure to thrive, and hyponatremia. The child was a previously well 8-month-old male who looked well but had mild periorbital edema with severe hypertension. After 3 days of captopril therapy, the nephrotic-range proteinuria significantly improved. However, the hypertension persisted. Renal imaging revealed a small left kidney with reduced parenchymal uptake and no significant excretion. A renal angiogram demonstrated left renal artery stenosis with increased left renal vein renin activity. The hypertension resolved within 24 h of a left nephrectomy, but non nephrotic-range proteinuria persisted for 8 months post operatively. Pathology of the left kidney was consistent with fibromuscular dysplasia. Although a few glomeruli (1%) had changes consistent with focal segmental glomerulosclerosis, such a few abnormal glomeruli were unlikely to account for the nephrotic syndrome. Hypertension-induced changes in the unaffected right kidney probably caused the nephrotic-range proteinuria. PMID- 12644924 TI - Nephrotic syndrome associated with human parvovirus B19 infection. AB - A previously healthy 8-year-old Japanese boy developed nephrotic syndrome during the course of erythema infectiosum due to human parvovirus B19 (PVB19) infection. A renal biopsy showed mesangiocapillary proliferative glomerulonephritis with immune complex deposits associated with PVB19 virus. His renal involvement improved spontaneously. PMID- 12644925 TI - Acute renal failure following deferoxamine overdose. AB - A 17-year-old patient with sickle cell-beta thalassemia undergoing treatment with home iron chelation therapy inadvertently received ten times the recommended dose of intravenous deferoxamine. Acute renal failure (ARF) developed within hours. Immediate treatment with high-efficiency hemodialysis resulted in the prompt return of renal function after only one hemodialysis session. No long-term nephrotoxic effects of the deferoxamine overdose developed after more than 1 year of follow-up. Children with sickle cell disease who are on intravenous deferoxamine and their parents should be cautioned about the possibility of ARF with overdose due to malfunction of the pump and/or inadequate monitoring during treatment. ARF, should it occur in such children, appears to respond well to treatment with high-efficiency hemodialysis. PMID- 12644926 TI - Cardiac ischemia during hemolytic uremic syndrome. AB - Increased thrombin generation and impaired fibrinolysis during Escherichia coli O157:H7-associated hemolytic uremic syndrome (HUS) plausibly diminish myocardial blood flow, but the frequency of cardiac ischemia during HUS is unknown. We identified a 9-year-old boy with HUS in whom myocardial diastolic dysfunction was demonstrated by echocardiography, who also had elevated serum troponin-I and creatine kinase MB mass. However, eight additional patients with HUS did not have elevated markers of cardiac injury. When present, elevated troponin-I should be considered to represent myocardial injury, and not attributed simply to renal insufficiency. It is possible that myocardial ischemia and secondary arrhythmias account for some sudden deaths that occur during acute HUS. PMID- 12644927 TI - Renal failure and acquired hypothyroidism. AB - The effects of thyroid metabolism on renal function in children are barely referred to in the literature. Primary hypothyroidism is known to be associated with a consistent elevation in serum creatinine levels. This is essentially because of the hypodynamic state that occurs in hypothyroidism, leading to a reduced glomerular filtration rate and hypercreatinemia. A teenager who developed renal failure due to primary hypothyroidism is reported. He displayed diverse serum biochemistry anomalies with an unremarkable physical examination. Thyroxine replacement therapy completely restored the euthyroid state and renal function. We propose, in accordance with other authors, measurement of thyrotropin levels in patients with hypercreatinemia. PMID- 12644928 TI - Renal, hepatic, and marrow dysfunction in a patient with chronic renal insufficiency. AB - Severe illness with acute renal failure, leukopenia, thrombocytopenia, and coagulopathy frequently occurs with hemolytic uremic syndrome/thrombotic thrombocytopenic purpura, hematological malignancies, sepsis, and collagen vascular diseases. We present a 16-year-old male fast-food worker with underlying chronic renal insufficiency who manifested these abnormalities as a result of Ehrlichia chaffeensis sepsis. Doxycycline therapy and aggressive supportive care led to complete recovery. PMID- 12644930 TI - Small bowel herniation and gangrene from peritoneal dialysis catheter exit site. AB - Peritoneal dialysis is one of the standard methods for blood purification. It is particularly well suited for treating children with acute renal failure. Here we report a rare case of small bowel herniation at the peritoneal catheter exit site following removal, leading to gangrenous infarction. PMID- 12644929 TI - End-stage renal failure in a child with X-linked ichthyosis. AB - We describe an 8-year-old boy who presented with steroid-resistant nephrotic syndrome (SRNS) associated with X-linked ichthyosis (XLI). At birth, the patient exhibited scaly skin, cryptorchidism, and steroid sulfatase (STS) deficiency. DNA analysis showed deletion of exons 1-10 of the STS gene. Proteinuria developed at 6 years and was resistant to steroid therapy. Kidney biopsy findings prior to steroid therapy were compatible with minimal change nephrotic syndrome. By immunofluorescence, glomerular basement membranes exhibited diffuse linear staining for the alpha5 chain of collagen IV, making X-linked Alport syndrome an unlikely explanation for the association of SRNS and ichthyosis. Despite immunosuppressive therapy together with oral prednisolone, no clinical response was achieved. He rapidly reached end-stage renal failure and finally underwent renal transplantation. We propose that SRNS should be considered as one of the highly variable phenotypes associated with XLI. PMID- 12644931 TI - Ataxia as a delayed reaction to contrast medium for an intravenous pyelography. PMID- 12644932 TI - Severe acute abdominal pain in idiopathic nephrotic syndrome. PMID- 12644933 TI - Renal tubular dysgenesis with medullary ray nodules. PMID- 12644934 TI - Acro-pectoro-renal field defect. PMID- 12644935 TI - Examination of sequence homology between human chromosome 20 and the mouse genome: intense conservation of many genomic elements. AB - The conservation of genomic organization of mammalian species has been of interest for its usefulness in characterizing the genetics of traits and diseases and as one tool for examining evolution. The recent rough draft sequencing of the mouse and human genomes provides the opportunity for more detailed analyses. The current study examines the extent of homology between human chromosome 20 and the mouse genome by comparing putative coding and non-coding sequence to provide insight into organizational and sequence similarities between the species. The relative position of each of 460 putative coding orthologues was the same in both species, except for a single genomic segment rearrangement. The similarity extended to exon/intron structure, the size of introns, as well as strong evidence for the conservation of position of ancient LINE-1, LINE-2 and LTR repetitive sequence and the subtelomeric region of the long arm of human chromosome 20 and that of mouse chromosome 2. There was also evidence for conservation of a limited amount of non-coding single-copy sequence. Together these data provide additional insight into the extent of conservation of mammalian genomic organization and sequence. PMID- 12644936 TI - Modulating action of the new polymorphism L436F detected in the GLB1 gene of a type-II GM1 gangliosidosis patient. AB - We report the modulating action of the L436F new polymorphism identified in the GLB1 gene of a patient affected by GM1 gangliosidosis with onset at 17 months and rapidly progressive psychomotor deterioration. Sequencing analysis and familial restriction studies revealed that the maternal allele of this patient carried the L436F polymorphism in cis with the known R201C mutation. The new mutation R68W was identified in his paternal allele. Since the GLB1 activity of the patient's leukocytes was very low and compatible with both the type-I and the type-II form of the disease, the potential impact of each mutation was investigated by expression studies in COS1 cells, and Western blots. Expression study of the R68W mutated allele resulted in no GLB1 activity. Transfection with a vector carrying the R201C mutation gave rise to a residual GLB1 activity, which, interestingly, was severely reduced in transfection with the L436F/R201C allele. These expression studies, together with co-transfection experiments, suggest that the R201C/L436F GLB1 "complex allele" leads to this patient's clinical and biochemical findings. The type-II phenotype of the disease is subdivided into late infantile and juvenile forms. The clinical and molecular characterization of this patient as late-infantile GM1 gangliosidosis is in keeping with a clear-cut division between the two sub forms of the type-II phenotype. The modulating role of the L436F polymorphism should be stressed as a cause of this patient's condition. This model suggests that the combination of missense mutations or polymorphisms should be evaluated when diagnosing inherited genetic disorders. PMID- 12644938 TI - Changes in macular thickness and depth of anterior chamber in patients after filtration surgery. AB - BACKGROUND: The prospective study was undertaken to analyze retinal thickness of the macula and anterior chamber depth in patients undergoing filtration surgery at different times of follow-up. METHODS: Forty-four patients (45 eyes) were included. The thickness of the fovea and the extrafoveal retina were measured using optical coherence tomography (OCT). Anterior chamber depth was determined using the IOLMaster. All standard clinical examination and measurements were performed prior to surgery, at the 2nd day (1-3 days), 1 week, 1 month and 3 months after surgery. RESULTS: Preoperative IOP under topical medication was 25.5+/-7.5 mmHg. IOP was significantly decreased at all postoperative visits ( P<0.0001): 10.8+/-5.9 mmHg at the 2nd day, 10.4+/-5.2 mmHg after 1 week, 11.7+/ 4.4 mmHg after 1 month and 11.4+/-3.6 mmHg after 3 months. No patient in our study suffered from prolonged postoperative hypotony (IOP<5 mmHg). Foveal thickness at the 2nd day, 1 week and 1 month were elevated compared with baseline ( P<0.004). Macular thickness changed from 164+/-20 microm before surgery to 168+/-20 microm after 2 days, 170+/-21 microm after 1 week, 173+/-19 microm after 1 month and 165+/-16 microm after 3 months. Mean thickness of the extrafoveal retina did not change significantly. Also, there were no significant changes in anterior chamber depth. There was no significant correlation between reduction of IOP and retinal thickness or anterior chamber depth. CONCLUSION: The reduction of IOP after filtration surgery leads to a moderate increase in foveal retinal thickness over approximately 1 month and is not associated with shallowing of the anterior chamber, provided that prolonged postoperative hypotony is avoided with the surgical technique used. PMID- 12644939 TI - Electronically measured compliance with occlusion therapy for amblyopia is related to visual acuity increase. AB - PURPOSE: We set out to determine whether the children who have low compliance (measured electronically) with occlusion therapy for amblyopia are those with insufficient increase of visual acuity. METHODS: In 14 newly identified amblyopic children (mean age 4.3+/-1.9 years), compliance was measured electronically over a period of 1 week, 6 months after the start of occlusion therapy. Compliance was measured with an Occlusion Dose Monitor (ODM). The measurements took place during planned domiciliary visits. The children were diagnosed with anisometropia (n=5), strabismus (n=4) and anisometropia and strabismus (n=5). Compliance was expressed in percentages of the electronically registered time compared with the prescribed occlusion time. Satisfactory acuity increase following 6 months of occlusion therapy was defined on reaching any of the following criteria: acuity increase expressed as a ratio between acuity of the amblyopic eye and acuity of the good eye of more than 0.75, acuity of the amblyopic eye exceeding 0.5 as measured on the E-Chart or Landolt-C, or three LogMAR lines of increase in acuity. RESULTS: Measured compliance averaged 80% in the eight children who had a satisfactory acuity increase and 34% in the six children who had an unsatisfactory visual acuity increase. Children with low acuity increase had statistically significantly lower compliance (P=0.038). CONCLUSION: The general assumption among orthoptists, that compliance with occlusion therapy for amblyopia is low in children with insufficient acuity increase, has been validated by electronic, objective means. PMID- 12644940 TI - Alterations of retinal capillary blood flow in preclinical retinopathy in subjects with type 2 diabetes. AB - BACKGROUND: To identify alterations of retinal capillary blood flow in the papillomacular area in preclinical diabetic retinopathy using the Heidelberg scanning laser Doppler flowmeter. METHODS: Ten eyes from ten patients with type 2 diabetes and no lesions visible on fundus photography (level 10 of Wisconsin grading) and ten eyes from ten healthy subjects of similar age range were examined with the HRF. Intravisit reproducibility of retinal capillary blood flow measurements was assessed in normal subjects and in type 2 diabetic patients, comparing different measurement areas and different analysis procedures: (a) 10x10 pixel box with original software, (b) 10x10 pixel box with SLDF software, and (c) whole-scan analysis with SLDF software (automatic full-field perfusion image analysis). RESULTS: Intravisit reproducibility for the whole-scan analysis in the papillomacular area was 3.52%, 4.81% and 4.60% for volume (VOL), flow (FLW) and velocity (VEL) respectively. Using this method, mean and SD values for retinal capillary blood-flow are 13.25+/-2.87, 214.58+/-55.30 and 0.74+/-0.17, for VOL, FLW and VEL for healthy eyes, comparing with 19.85+/-6.22, 360.87+/ 158.70 and 1.20+/-0.48 in eyes with preclinical diabetic retinopathy (P<0.010, P<0.019 and P<0.015 respectively). CONCLUSIONS: The HRF shows acceptable reproducibility when using whole-scan analysis in the papillomacular area. Retinal capillary blood VOL, FLW and VEL were particularly increased in five of the ten diabetic eyes examined, with values over the mean + 2SD of the control population, suggesting that eyes showing increased retinal capillary blood flow may indicate risk of progression. PMID- 12644941 TI - Prognostic value of Hutchinson's sign in acute herpes zoster ophthalmicus. AB - PURPOSE: To determine the prognostic value of nasociliary skin lesions (Hutchinson's sign) for ocular inflammation and corneal sensory denervation in acute herpes zoster ophthalmicus. METHODS: A longitudinal observational study with a 2-month follow-up was performed involving 83 non-immunocompromised adults with acute herpes zoster ophthalmicus, with a skin rash duration of less than 7 days, referred by their general practitioner. All skin lesions at the tip, the side and the root of the nose, representing the dermatomes of the external nasal and infratrochlear branches of the nasociliary nerve, were documented by taking photographs and marking anatomical drawings. Ocular inflammatory signs were observed by slit-lamp biomicroscopy, and corneal sensitivity was measured with the Cochet-Bonnet esthesiometer at 2-month follow-up. RESULTS: Hutchinson's sign was a powerful predictor of ocular inflammation and corneal denervation in herpes zoster ophthalmicus [relative risks: 3.35 (CI 95%: 1.82-6.15) and 4.02 (CI 95%:1.55-10.42), respectively]. The manifestation of herpes zoster skin lesions at the dermatomes of both nasociliary branches was invariably associated with the development of ocular inflammation. CONCLUSION: Clinicians should be alert for early skin lesions within the complete nasociliary dermatome, because they are a reliable prognostic sign of sight-threatening ocular complications in acute herpes zoster ophthalmicus. PMID- 12644942 TI - Preservative-free diclofenac sodium 0.1% for vernal keratoconjunctivitis. AB - PURPOSE: The aim of this study is to evaluate the efficacy of prolonged treatment with preservative-free diclofenac sodium 0.1% eye drops in patients with vernal keratoconjunctivitis (VKC). METHODS: A prospective open study was performed in 22 patients with VKC treated with preservative-free diclofenac sodium 0.1% eye drops. Patients used the eye drops four times daily in both eyes for 120 days. Signs (papillae, hyperaemia and corneal lesions) and symptoms (itching, redness and photophobia) of the ocular surface were graded and statistically evaluated before and after treatment by a non-parametric test (Mann-Whitney U-test). RESULTS: Forty per cent of the patients showed an improvement in their symptoms at the end of the treatment. Total signs and symptoms scores were significantly decreased at the end of treatment compared with the baseline values (from 6.13+/ 1.45 to 0.81+/-0.90 and from 5.40+/-1.18 to 2.63+/-0.95, respectively; P<0.001). Significant decreases in conjunctival redness (P<0.001), itching (P<0.001) and photophobia (P<0.001 ) were observed at the end of treatment. Conjunctival hyperaemia was significantly reduced (P<0.001) at the end of treatment, while no significant differences were observed for corneal lesions and for papillary size. No patient showed exacerbation of the disease during the treatment. CONCLUSION: VKC is a chronic disease that requires prolonged treatment to control the inflammatory process. Our preliminary study demonstrates the efficacy and safety of preservative-free diclofenac sodium 0.1% eye drops in controlling the signs and symptoms of VKC in prolonged treatment. PMID- 12644943 TI - Results of the betaxolol versus placebo treatment trial in ocular hypertension. AB - PURPOSE: To determine whether treatment with betaxolol can delay or prevent the conversion from ocular hypertension to early glaucoma on the basis of visual field criteria, by means of a prospective, randomised, placebo-controlled trial. METHODS: Three hundred and fifty-six ocular hypertensives were randomized to treatment with either betaxolol drops or placebo drops during the period 1992 1996. Each patient was followed prospectively with 4-monthly visits. Examination at each visit included visual field testing, intra-ocular pressure (IOP) measurement and optic disc imaging. Conversion to early glaucoma was defined on the basis of visual field change by AGIS criteria. An intent-to-treat analysis compared visual field conversion after 3 years in the treatment and placebo arms. Normal visual field survival analysis was also performed. The IOP characteristics of the two treatment groups were compared. RESULTS: Two hundred and fifty-five patients completed the study, which ended in 1998, with a range of follow-up of 2 6 years. Sixteen (13.2%) of 121 patients in the placebo group converted to glaucoma, compared with 12 (9.0%) of 134 patients in the betaxolol group. The intent-to-treat analysis demonstrated no evidence of any difference in conversion rates between the betaxolol and placebo groups after 3 years. Visual field survival analysis demonstrated no significant difference between the betaxolol and placebo groups. The betaxolol-treated group had significantly lower post treatment IOP values. Converters had significantly higher pre- and post-treatment IOP values than non-converters. CONCLUSIONS: Betaxolol significantly lowered the IOP level compared with placebo. Conversion to glaucoma was found to be related to both the baseline and post-treatment IOP levels. However the intent-to-treat analysis did not demonstrate a statistically significant reduction in the conversion rate in the betaxolol-treated group. PMID- 12644944 TI - Indocyanine green-assisted peeling of the epiretinal membrane in proliferative vitreoretinopathy. AB - BACKGROUND: We stained the internal limiting membrane of patients suffering from proliferative vitreoretinopathy with indocyanine green solution during proliferative vitreoretinopathy surgery to improve the visibility of the membranes, and thereby histopathologically confirmed the excised epiretinal membranes. METHODS: Three patients underwent a standard three-port pars plana vitrectomy with indocyanine green staining. After performing a subtotal vitrectomy we spread 0.5% indocyanine green solution, approximately 1 ml, on the retinal surface and peeled off the epiretinal membranes. RESULTS: The epiretinal membranes did not stain clearly, while the internal limiting membranes did stain clearly. We could therefore distinguish the epiretinal membranes from the retina. We cut the internal limiting membrane, grasped it, and peeled off the internal limiting membrane underlying the epiretinal membranes using vitreoretinal forceps. A histopathologic examination confirmed the presence of proliferative cells and an extracellular matrix underlying the internal limiting membranes. CONCLUSION: The technique for staining the epiretinal membranes in proliferative vitreoretinopathy using indocyanine green gives better visualization and allows surgeons to remove the epiretinal membranes more safely and effectively, as well as with less risk of retinal damage. PMID- 12644945 TI - Posterior capsule morphology determinants of visual function. AB - BACKGROUND: This study explores the relative influence of fibrosis and pearl-type posterior capsule opacification (PCO) on visual function. It also examines the relevance of analyzing different central areas of posterior capsule on visual function. METHODS: Thirty-eight eyes of 33 patients had their vision tested and posterior capsules photographed digitally. Each patient then underwent Nd:YAG laser capsulotomy. Patients returned 1 week later for repeat vision tests and photography. Analyses of areas of pearls and fibrosis were performed using the EPCO software system. The difference in these values before and after Nd:YAG laser capsulotomy was compared to change in vision for each patient. The effect of analyzing the central 1.5-, 2.5- and 3.5-mm zones was studied. RESULTS: Stepwise multiple linear regression analysis shows that improvements in distance and near acuity are most strongly related to the central 1.5-2.5 mm of fibrosis and the central 3.5 mm of pearls. Improvements in contrast sensitivity are most strongly related to the central 1.5 mm of both fibrosis and pearls. CONCLUSION: In the assessment of PCO, the location and morphology of opacification correlate with visual function. Central fibrosis and paraxial pearls are important determinants of visual dysfunction. PMID- 12644946 TI - Graded sensitiveness of the various retinal neuron populations on the glyoxal mediated formation of advanced glycation end products and ways of protection. AB - BACKGROUND: The accumulation of advanced glycation end products (AGEs) in retinal cells is known to be associated with the risk of diabetic retinopathy. To develop a model of AGE-related metabolic stress in retinal organ cultures, we investigated the accumulation of a typical glycoxidation product (N(epsilon) [carboxymethyl] lysine [CML]) and its possible pro-apoptotic effects on different retinal cell populations. METHODS: Retinal organ cultures (rat) were kept for 9 h in the Ames medium containing 0 (control), 5, 25, 50, 150, 300 and 800 micro M glyoxal. The expression of bax, active caspase-3, and the accumulation of CML were studied by using immunohistochemistry after the paraffin embedding of retinal explants. Apoptosis was studied using the terminal deoxynucleotidyl transferase-mediated dUTP digoxigenin nick end labeling (TUNEL) test and electron microscopy. Alpha lipoic acid (alpha-LA), sodium metavanadate (NaVO(3)), N acetylcysteine (NAC), aminoguanidine (AG), and nicotinamide (NA) were used to influence glyoxal effects in organ cultures. RESULTS: In cultured normal non diabetic retinae, small amounts of CML and the apoptosis-promoting factors bax and active caspase-3 were present. CML, bax and active caspase-3 increased after incubation with glyoxal. Incubation with glyoxal (<300 micro M, 9 h) increased apoptotic events in all layers. At low glyoxal concentrations, we found a graded sensitiveness of the different layers: at 25 micro M 39.4% in GCL, 28.2% in INL, 11.9% in ONL. After 800 micro M glyoxal, approximately 50% of the cells in all layers of the retina were apoptotic. In the ONL, this ratio was reduced by NaVO(3) (17%), by AG (27%), by NA (24.8%), by NAC (25.2%), and by alpha-LA (33.5%). In the INL, AG (25.9%) produced the best result. In the GCL, NAC, NaVO(3) and AG reduced apoptosis. A-LA had no significant protective effect. CONCLUSION: The glyoxal-induced rapid formation of CML shows the ability of our retina model to simulate AGE-related effects in vitro. The dose-dependent expression of apoptosis-promotor molecules indicates that the apoptosis-inducing machinery starts in most retinal cells within 9 h. The neurotoxicity of glyoxal induced AGE formation was shown by the significantly increased rate of cell death in the retina. The significant decrease of apoptotic events (P<0.01) indicates that antioxidants and AGE formation blocker can exert a differentiated cytoprotection for each of the retinal cell layers. PMID- 12644947 TI - Muscarinic blockers potentiate beta-adrenergic relaxation of bovine iris sphincter. AB - BACKGROUND: beta-Adrenergic relaxation of iris sphincter has been described in many species, including human. It is generally accepted that the size of the pupil is mainly controlled by muscarinic and alpha-adrenergic tonus. This study was undertaken to investigate the interactions between muscarinic and beta adrenergic drugs in the relaxation of bovine iris sphincter. METHODS: Intact bovine iris sphincters were incubated in an organ bath and challenged with appropriate beta-adrenergic agonists and/or muscarinic antagonists. Tension was measured by a force transducer. RESULTS: Isolated bovine iris sphincter responded to muscarinic stimulation by contraction and to beta-adrenergic stimulation by relaxation. The beta-adrenergic agonists isoproterenol (ISO) and salbutamol (SAL) each caused marked relaxation of sphincters pre-contracted with the muscarinic agonist carbamylcholine. Sphincters pre-treated with the muscarinic antagonists atropine (0.1 micro M) or ipratropium bromide (10 micro M) and challenged 5 min later with either isoproterenol (0.3 nM) or salbutamol (10 nM) exhibited approximately twofold potentiation of beta-adrenergic relaxation. Adding the drugs in the reverse order did not produce any potentiation over the effect of beta-adrenergic stimulation alone. The dose-response curve to isoproterenol in naive sphincters or sphincters pre-treated with atropine indicated that muscarinic blockade decreased the EC(50) rather than potentiated maximal beta adrenergic relaxation. CONCLUSION: Muscarinic antagonists potentiate beta adrenergic relaxation of the sphincter by affecting the apparent potency of beta adrenergic agonists and not by antagonizing the intrinsic tonus produced by endogenously released acetylcholine. PMID- 12644948 TI - Loss of RPE phenotype affects phagocytic function. AB - BACKGROUND: Cultured retinal pigment epithelial (RPE) cells exposed to vitreous lose their cuboidal epithelial phenotype and transform to a fibroblast-like morphology. This study investigates whether the loss of RPE phenotype will affect the ability of RPE to phagocytize photoreceptor outer segments (ROS). METHODS: First-passage porcine RPE cells were grown to confluence and exposed to an equal mixture of freshly homogenized vitreous and DMEM (10% FBS) for 48 h to allow transformation to occur. The phagocytic ability of normal epithelial and vitreous transformed RPE cultures was evaluated by determining the number of ROS bound and ingested. The expression and distribution of alphavbeta5 integrin in control and vitreous-treated RPE were examined by immunoprecipitation coupled with immunoblotting and immunohistochemistry and the contribution of alphavbeta5 integrin to ROS phagocytosis was evaluated using a functional blocking antibody. RESULTS: Bound ROS on vitreous-transformed RPE decreased to 33% of control and ingested ROS decreased to 47%. The expression of alphavbeta5 integrin was decreased significantly following vitreous transformation as compared with controls and the immunolocalization of alphavbeta5 on control RPE was restricted to apical patches while on vitreous-treated RPE, labeling was diminished and randomly dispersed. Five times less blocking antibody was required to attain maximal phagocytic inhibition in these cultures, but some phagocytic activity remained in both normal and transformed cultures even at saturating concentrations. CONCLUSION: The decrease in ROS phagocytosis observed in transformed RPE correlated with the down- regulation of alphavbeta5 integrin expression and its random distribution on the RPE. PMID- 12644949 TI - von Hippel-Lindau disease type 2A in a family with a duplicated 21-base-pair in frame insertion mutation in the VHL gene. AB - BACKGROUND: von Hippel-Lindau disease (VHL), also called angiomatosis retinae, is inherited as an autosomal dominant trait. It is frequently associated with other tumors in the central nervous system, kidneys, or adrenal glands. In order to investigate the relationship between genotype and corresponding phenotypes, we performed molecular genetic analysis in a Japanese patient with VHL type 2A. METHODS: After informed consent had been obtained, the three exons of the VHL gene were PCR-amplified and sequenced either directly or after subcloning. Clinical features were also examined. RESULTS: A novel in-frame duplication of the 21 base pairs at nucleotide 806 (the position of codon 198) of the VHL gene was found in our patient. The clinical phenotype of the patient included retinal hemangiomas associated with vitreous hemorrhage and traction retinal detachment, pheochromocytoma, and hemangioma-like lesions in the cerebellum which corresponded to those of VHL type 2A. Abnormal diffuse vascular leakage was observed in the apparently intact retina by fluorescein angiography. CONCLUSION: An insertion mutation of the VHL gene is a rare association with VHL type 2. This insertion mutation may interfere the binding between the VHL gene and elongins. Abnormal retinal vascular leakage suggests the possible effects of overexpressed vascular permeability factors such as vascular endothelial growth factor from hemangiomas associated with defective VHL gene. PMID- 12644950 TI - Neovascular ingrowth site photothrombosis in choroidal neovascularization associated with retinal pigment epithelial detachment. AB - PURPOSE: To describe the clinical, angiographic and optical coherence tomography (OCT) findings of patients with choroidal neovascularization (CNV) associated with retinal pigment epithelial detachment (PED) who were treated by neovascular ingrowth site photothrombosis. METHODS: Interventional case reports. After identification of the CNV feeder vessels, two patients had focal photothrombosis of the neovascular ingrowth site using continuous 810-nm laser application after intravenous indocyanine green (ICG) injection and were prospectively followed with fluorescein and ICG angiography as well as OCT. RESULTS: Both patients presented an increase of 3 or more ETDRS lines in visual acuity by 12 months of follow-up. Fluorescein and ICG angiography demonstrated reduced lesion perfusion and regression of the PEDs; accordingly, OCT disclosed decrease in retinal thickness and elevation caused by rapid fluid resolution. There were no significant complications related to the procedure. CONCLUSION: Photothrombosis using 810-nm light to direct laser energy continuously at the neovascular ingrowth site after intravenous ICG infusion is effective in restoring macular architecture and improving vision in patients with CNV complicated by PEDs by means of substantial decrease in the lesion perfusion and resolution of exudative manifestations. PMID- 12644951 TI - Developmentally regulated extrachromosomal circular DNA formation in the mesozoan Dicyema japonicum. AB - The dicyemid mesozoans are simple multicellular parasites with a long cylindrical axial cell surrounded by a single outer layer of 20 to 30 ciliated peripheral somatic cells. Their larval development proceeds within the axial cell. Here we demonstrate the appearance of extrachromosomal circular DNAs and their fate during early embryogenesis in Dicyema japonicum. These DNAs are highly heterogeneous in sequence, suggesting that they consist of unique--not repetitive -elements. Potential open reading frames were not evident in the elements, so these DNAs are unlikely to have a protein-encoding function. In situ hybridization revealed that the circular DNA elements were restricted to the early embryonic larvae and gradually faded out as larvae approached maturity. Furthermore Southern blot analysis and polymerase chain reaction analysis using a high molecular weight DNA as a template provided evidence that the extrachromosomal DNA circles are originally present in chromosomes. These observations suggest DNA elimination--or selective replication--of the elements from chromosomes during early embryogenesis in dicyemid mesozoans. PMID- 12644953 TI - Influence of the SuUR gene on intercalary heterochromatin in Drosophila melanogaster polytene chromosomes. AB - Salivary gland polytene chromosomes of Drosophila melanogaster have a reproducible set of intercalary heterochromatin (IH) sites, characterized by late DNA replication, underreplicated DNA, breaks and frequent ectopic contacts. The SuUR mutation has been shown to suppress underreplication, and wild-type SuUR protein is found at late-replicating IH sites and in pericentric heterochromatin. Here we show that the SuUR gene influences all four IH features. The SuUR mutation leads to earlier completion of DNA replication. Using transgenic strains with two, four or six additional SuUR(+) doses (4-8xSuUR(+)) we show that wild type SuUR is an enhancer of DNA underreplication, causing many late-replicating sites to become underreplicated. We map the underreplication sites and show that their number increases from 58 in normal strains (2xSuUR(+)) to 161 in 4 8xSuUR(+) strains. In one of these new sites (1AB) DNA polytenization decreases from 100% in the wild type to 51%-85% in the 4xSuUR (+) strain. In the 4xSuUR(+) strain, 60% of the weak points coincide with the localization of Polycomb group (PcG) proteins. At the IH region 89E1-4 (the Bithorax complex), a typical underreplication site, the degree of underreplication increases with four doses of SuUR(+) but the extent of the underreplicated region is the same as in wild type and corresponds to the region containing PcG binding sites. We conclude that the polytene chromosome regions known as IH are binding sites for SuUR protein and in many cases PcG silencing proteins. We propose that these stable silenced regions are late replicated and, in the presence of SuUR protein, become underreplicated. PMID- 12644952 TI - Mammalian meiosis involves DNA double-strand breaks with 3' overhangs. AB - Meiotic recombination in yeast is initiated at DNA double-strand breaks (DSBs), processed into 3' single-strand overhangs that are active in homology search, repair and formation of recombinant molecules. Are 3' overhangs recombination intermediaries in mouse germ cells too? To answer this question we developed a novel approach based on the properties of the Klenow enzyme. We carried out two different, successive in situ Klenow enzyme-based reactions on sectioned preparations of testicular tubules. Signals showing 3' overhangs were observed during wild-type mouse spermatogenesis, but not in Spo11(-/-) males, which lack meiotic DSBs. In Atm(-/-) mice, abundant positively stained spermatocytes were present, indicating an accumulation of non-repaired DSBs, suggesting the involvement of ATM in repair of meiotic DSBs. Thus the processing of DSBs into 3' overhangs is common to meiotic cells in mammals and yeast, and probably in all eukaryotes. PMID- 12644954 TI - Nucleolin provides a link between RNA polymerase I transcription and pre-ribosome assembly. AB - Despite the identification of numerous factors involved in ribosomal RNA synthesis and maturation, the molecular mechanisms of ribosome biogenesis, and in particular the relationship between the different steps, are still largely unknown. We have investigated the consequences of an increased amount of a major nucleolar non-ribosomal protein, nucleolin, in Xenopus laevisstage VI oocytes on the production of ribosomal subunits. We show that a threefold increase in nucleolin leads to the complete absence of pre-rRNA maturation in addition to significant repression of RNA polymerase I transcription. Observation of "Christmas trees" by electron microscopy and analysis of the sedimentation properties of 40S pre-ribosomal particles suggest that an increased amount of nucleolin leads to incorrect packaging of the 40S particle. Interestingly, nucleolin affects the maturation of the 40S particle only when it is present at the time of transcription. These results indicate that nucleolin participates in the co-transcriptional packaging of the pre-rRNA, and that the quality of this packaging will determine whether the 40S precursor undergoes maturation or is degraded. The interaction of nucleolin with nascent pre-rRNA could help the co transcriptional assembly on pre-rRNA of factors necessary for the subsequent maturation of the pre-ribosomal particle containing the 40S pre-rRNA. PMID- 12644956 TI - Pulmonary hypertension. AB - Pulmonary arterial hypertension (PAH) must be classified into primary pulmonary hypertension and PAH related to other diseases such as collagen vascular diseases, HIV infection or portal hypertension. PAH must also be differentiated from other entities, in particular pulmonary hypertension secondary to thromboembolic diseases, requiring specific approaches. All PAH results in similar histological remodelling of pulmonary arteries, with thickening of the intima, proliferation of the media and plexogenic lesions. Today the physiopathology of these lesions is much better understood and has resulted in new therapies involving substances such as prostacyclins, endothelin receptor antagonists or phosphodiesterase inhibitors, aimed not only at dilating arteries but also at preventing their remodelling. Thromboendarterectomy, septostomy and transplantation remain the only option where medical treatment has failed. PMID- 12644955 TI - Rye terminal neocentromeres: characterisation of the underlying DNA and chromatin structure. AB - We have studied rye plants with neocentromeres on the terminal regions of the chromosomes. These neocentromeres only appear in meiosis, they are active together with the normal centromere and move the chromosomal arms polewards from prometaphase to anaphase at both the first and second meiotic divisions. All chromosomes of the normal set may show neocentric activity, but chromosomal arms with terminal heterochromatic blocks, as assessed by C-banding, are significantly more susceptible than those that do not have them. At least three repetitive sequences underlie the neocentromeres: pSc34, pSc74 and pSc200. These sequences are not detectable in B chromosomes, which never showed neocentric activity. Fluorescence in situ hybridisation with these sequences used as probes revealed elongated chromatin extensions on the neocentromeres that have not been observed using other staining techniques. These extensions were never observed in control plants. They suggest a modified chromatin structure, which might be responsible for the interaction with proteins involved in chromosomal movement on the spindle. PMID- 12644957 TI - Update on cystic fibrosis: selected aspects related to lung transplantation. AB - Survival after lung transplantation for cystic fibrosis has improved substantially. To date, 1-year survival is as much as 80-90% and 5-year survival 60-70%. Experience of surgical techniques, periand postoperative management and long-term follow-up care has grown. High risk patients for whom transplantation was contraindicated ten years ago are today being transplanted. Prerequisites for a successful outcome are appropriate timing of referral, careful consideration of medical issues in other organs, and psychosocial support systems. Panresistant organisms are a special problem in candidates with cystic fibrosis, and advances in microbiological testing and characterisation of these organisms are warranted. Living donor lobar transplantation has become an option in rapidly deteriorating children and young adults. Selected aspects of the evolving field of lung transplantation are discussed. PMID- 12644958 TI - The efficacy of flumazenil in subclinical to mild hepatic encephalopathic ambulatory patients. A prospective, randomised, double-blind, placebo-controlled study. AB - OBJECTIVES: Hepatic encephalopathy (HE) is a neuropsychiatric syndrome associated with fulminant hepatic failure and chronic liver disease. Its pathogenesis is unclear. One of the factors implicated is enhanced GABA-ergic tone, which is probably related to increased concentrations of cerebral benzodiazepine (BNZ). In the present study, we tested flumazenil, a cerebral BNZ antagonist, in cirrhosis patients with hepatic encephalopathy. METHODS: Out of 47 patients, 7 were excluded prior to randomization for various reasons. Twenty patients were included in the flumazenil group and 20 in the placebo group in a prospective, randomized, double-blind, placebo-controlled study. Patients were given flumazenil (1 mg/h, continuous IV infusion) or an equal volume of saline solution for 5 hours. Before and after treatment, portosystemic encephalopathy (PSE) stage and number connection test (NCT) scores were checked every half hour for 5 hours. EEG was recorded 15 minutes before and 1 hour after treatment. RESULTS: While significant improvements were determined in PSE stage and NCT score in the flumazenil group, there were no such improvements in the placebo group. There was no statistically significant difference between pre- and post-treatment EEGs in either group. CONCLUSION: It was concluded that continuous IV infusion of flumazenil had beneficial and safe effects in the treatment of hepatic encephalopathy patients. PMID- 12644960 TI - Atypical antipsychotics in the treatment of schizophrenia. AB - Over the last few years atypical antipsychotics have been used increasingly in the pharmacological treatment of schizophrenia. This review provides an overview of the pharmacological and clinical characteristics of atypical antipsychotics. In particular we discuss their efficacy in treating psychotic and negative symptoms as well as treatment-refractory patients. The most important side effects as well as their possible interactions are reviewed in detail. Recommendations for the clinical use are given. PMID- 12644959 TI - High risk behaviour and fertility desires among heterosexual HIV-positive patients with a serodiscordant partner--two challenging issues. AB - AIM OF THE STUDY: To evaluate fertility intentions and condom use among HIV positive persons. METHODS: Multicentre study based on anonymous data collection (questionnaire). RESULTS: 114 questionnaires providing complete information were evaluated. 45% of HIV-positive women and 38% of HIV-positive men expressed the desire for children. Irrespective of this wish, half the study participants felt that health care providers would not sufficiently address their concerns regarding relationship, sexuality and fertility intentions. In HIV-discordant heterosexual couples, consistent condom use was mentioned by 73% of respondents. Among study participants no significant relationship between HAART, viral load and inconsistent condom use was found. In contrast, information on condom use obtained from the Swiss HIV Cohort Study (SHCS), where the information is obtained by interview, gave higher estimates (88%) of consistent condom use. CONCLUSIONS: A significant proportion of HIV-infected individuals express a wish for parenthood. Issues related to fertility intentions and sexual relations need to be addressed more frequently by health care providers. Non-anonymous data collection on condom use may underestimate high risk behaviour. PMID- 12644961 TI - Thalidomide: from tragedy to promise. AB - Thalidomide is an immunomodulatory and antiangiogenic drug. Although the exact mechanism of action is not fully understood, it has been shown to be active in a variety of diseases. There are multiple trials going on to evaluate the optimal dose of thalidomide and the importance of combining thalidomide with other drugs. This review introduces the properties and putative mechanism of action of thalidomide and summarizes the most important clinical trials with this biological modifier. PMID- 12644963 TI - Two years outcome of very pre-term and very low birthweight infants in Switzerland. AB - BACKGROUND: There are only few reports worldwide on the outcome of very pre-term infants and very low birthweight infants for a whole country. In Switzerland official population statistics are based on birthweight only, gestational age not yet being documented. AIM: The aim of the present study was to assess the outcome at two years of age for a geographically defined high-risk neonatal population based on both birthweight and gestational age. METHODS: All infants born in 1996 included in the Swiss Neonatal Network (a national anonymous registry established by the Swiss Society of Neonatology for liveborn infants before 32 completed gestational weeks or weighing less than 1500 g) were divided into three groups according to gestational age and birth weight: Group 1: born <32 completed gestational weeks and weighing =1500 g; group 2: born after 32 completed gestational weeks and weighing <1500 g; group 3: born <32 gestational weeks and weighing <1500 g. Information at 24 months corrected age about growth, neurological outcome, frequency of respiratory infections, prescription of antibiotics and medical consultations during this period was obtained from the paediatricians caring for the infants. Fair outcome was defined as survival without serious neonatal complications or abnormal neurological findings at 24 months corrected for prematurity. RESULTS: 723 infants were born alive in Switzerland between 1.1. and 31.12.1996 before 32 completed weeks or weighing less than 1500 g at birth. Mortality was 4.3% for a total of 163 infants in group 1 (<32 weeks, =1500 g), 4.6% for 108 infants in group 2 (>32 weeks, <1500 g) and 18.6% for 452 infants in group 3 (<32 weeks, <1500 g). 6.5% of group 1 survivors followed up to 24 months corrected age had a poor neurological outcome as compared to 9.3% in group 2 and 10.9% in group 3. Infants in group 1 needed antibiotics less often after hospital discharge (interquartile range IQR: 0-2 courses) than infants in group 2 (0-3 courses) and 3 (0-3 courses). Infants in group 2 suffered from fewer airway infections (interquartile range 2-5 times) than in group 1 (2-6 times) and 3 (1-7 times). Infants in group 3 needed more medical consultations (IQR 12-21) than those in group 1 (10-16) and 2 (11-16). The overall fair outcome at 24 months corrected age was 85.3% in group 1, 80.7% in group 2 and 59.6% in group 3. A close correlation between overall fair outcome and gestational age at birth on the one hand and with birthweight on the other can be observed. CONCLUSIONS: This study gives estimates for mortality, poor and fair outcome at 24 months corrected age for very low birth weight infants (<1500 g) and for very pre-term infants (<32 completed gestational weeks). Gestational age is as important for predicting outcome as birthweight and should therefore be integrated into national statistics. PMID- 12644962 TI - National survey on prescription of cardiovascular drugs among outpatients with coronary artery disease in Switzerland. AB - BACKGROUND: Secondary prevention of coronary artery disease markedly reduces cardiovascular mortality and non-fatal endpoints. Outpatient care of subjects with coronary artery disease has been assessed in several European countries, but no current data is available for Switzerland. METHODS: A random sample of office based physicians across Switzerland recorded current drug prescription of outpatients with coronary artery disease in the years 2000/2001 by means of a mail questionnaire. We assessed treatment frequencies according to different patient characteristics. RESULTS: 565 patients were included (mean age 68 +/- 11 years, 75% male). There was no evidence for differences in drug utilisation among the regions. Drug prescription rates for antithrombotic agents, beta-blockers, ACE-inhibitors/angiotensin receptor blockers and lipid lowering drugs were 91%, 58%, 50% and 63% respectively. Lower treatment rates were observed among patients >70 years and in those without a history of myocardial infarction or coronary revascularisation. Forty-nine percent of the patients had a blood pressure >140/>90, and 60% had lipid readings above the intervention cut-off according to the Swiss recommendations. Among those without a history of myocardial infarction or coronary revascularisation, the respective figures were 60% and 80%. CONCLUSIONS: Compared to former surveys evidence based drug prescription has improved in Switzerland. Despite this, therapeutic goals for cholesterol levels and blood pressure are not being reached in a large proportion of patients. A high risk group for under use of evidence based drugs are patients without a history of myocardial infarction or coronary revascularisation. PMID- 12644964 TI - Zopiclone: some remarks on pharmacokinetics and pharmacodynamics. PMID- 12644965 TI - Estimation and testing of parent-of-origin effects for quantitative traits. AB - Recent progress in developing family-based association methods has extended their use to the analysis of quantitative traits in the offspring and to the estimation, for dichotomous traits, of the relative contribution of genetic and environmental mechanisms for parent-of-origin effects. However, many traits of interest are not naturally measured on a binary scale yet are suspected or known to be influenced by imprinted genes, and there is consequent interest in seeking evidence for parent-of-origin effects at these loci. Here we show how simple linear models can be used to estimate these parent-of-origin effects for a broad class of phenotypes; in particular, normally distributed quantitative traits are easily dealt with. PMID- 12644966 TI - Native American Y chromosomes in Polynesia: the genetic impact of the Polynesian slave trade. AB - Since Thor Heyerdahl asserted that Polynesia was first colonized from the Americas (Heyerdahl 1950), geneticists have sought--but have not found--any evidence to support his theories. Here, Native American Y chromosomes are detected on the Polynesian island of Rapa. However, this, together with other odd features of the island's Y-chromosomal gene pool, is best explained as the genetic impact of a 19th century Peruvian slave trade in Polynesia. These findings underscore the need to account for history before turning to prehistory and the value of archival research to understanding modern genetic diversity. Although the impact of the Atlantic slave trade on the distribution of modern genetic diversity has been well appreciated, this represents the first study investigating the impact of this underappreciated episode on genetic diversity in the Pacific. PMID- 12644967 TI - Genetic linkage analysis of a dichotomous trait incorporating a tightly linked quantitative trait in affected sib pairs. AB - Many complex diseases are usually considered as dichotomous traits but are also associated with quantitative biological markers or quantitative risk factors. For such dichotomous traits, although their associated quantitative traits may not directly underly the diagnosis of the disease status, if the associated quantitative trait is also linked to the chromosomal regions linked to the dichotomous trait, then joint analysis of dichotomous and quantitative traits should be more efficient than consideration of them separately. Previous studies have focused on the situation when a dichotomous trait can be modeled by a threshold process acting on a single underlying normal liability distribution. However, for many complex disorders, including most psychiatric disorders, diagnosis is generally based on a set of binary or discrete criteria. These traits cannot be modeled on the basis of a threshold process acting on an underlying continuous trait. We propose a likelihood-based method that efficiently combines such a discrete trait and an associated quantitative trait in the analysis, using affected-sib-pair data. Our simulation studies suggest that joint analysis increases the power to detect linkage of dichotomous traits. We also apply the proposed new method to an asthma genome-scan data set and incorporate the total serum immunoglobulin E level in the analysis. PMID- 12644970 TI - Overview of the diabetic foot. PMID- 12644971 TI - Diabetic vascular disease and its management. PMID- 12644968 TI - Missense mutations in the regulatory domain of PKC gamma: a new mechanism for dominant nonepisodic cerebellar ataxia. AB - We report a nonepisodic autosomal dominant (AD) spinocerebellar ataxia (SCA) not caused by a nucleotide repeat expansion that is, to our knowledge, the first such SCA. The AD SCAs currently comprise a group of > or =16 genetically distinct neurodegenerative conditions, all characterized by progressive incoordination of gait and limbs and by speech and eye-movement disturbances. Six of the nine SCAs for which the genes are known result from CAG expansions that encode polyglutamine tracts. Noncoding CAG, CTG, and ATTCT expansions are responsible for three other SCAs. Approximately 30% of families with SCA do not have linkage to the known loci. We recently mapped the locus for an AD SCA in a family (AT08) to chromosome 19q13.4-qter. A particularly compelling candidate gene, PRKCG, encodes protein kinase C gamma (PKC gamma), a member of a family of serine/threonine kinases. The entire coding region of PRKCG was sequenced in an affected member of family AT08 and in a group of 39 unrelated patients with ataxia not attributable to trinucleotide expansions. Three different nonconservative missense mutations in highly conserved residues in C1, the cysteine-rich region of the protein, were found in family AT08, another familial case, and a sporadic case. The mutations cosegregated with disease in both families. Structural modeling predicts that two of these amino acid substitutions would severely abrogate the zinc-binding or phorbol ester-binding capabilities of the protein. Immunohistochemical studies on cerebellar tissue from an affected member of family AT08 demonstrated reduced staining for both PKC gamma and ataxin 1 in Purkinje cells, whereas staining for calbindin was preserved. These results strongly support a new mechanism for neuronal cell dysfunction and death in hereditary ataxias and suggest that there may be a common pathway for PKC gamma related and polyglutamine-related neurodegeneration. PMID- 12644972 TI - Diabetic revascularization: improving limb salvage in the absence of autogenous vein. PMID- 12644973 TI - Diabetic neuropathy. PMID- 12644974 TI - Biomechanical impact on the treatment of neuropathic ulcerations. PMID- 12644975 TI - Maximizing length and optimizing biomechanics in foot amputations by avoiding cookbook recipes for amputation. PMID- 12644976 TI - Application of external fixators for management of Charcot deformities of the foot and ankle. PMID- 12644977 TI - Nonoperative management of diabetic foot infections. PMID- 12644979 TI - The epidermal growth factor receptor-tyrosine kinase: a promising therapeutic target in solid tumors. AB - The overexpression and aberrant function of the epidermal growth factor receptor (EGFR) and its ligands in several human carcinomas have provided a rationale for targeting this signaling network with novel treatment approaches. The epidermal growth factor receptor-tyrosine kinase (EGFR-TK) is a selective target for inhibiting cancer because it is activated in many tumor cells, yet is strictly controlled in normal cells. The EGFR-TK initiates diverse signal transduction pathways in tumor cells that have a profound effect on their biology. Activation of the EGFR-TK provides signals that drive dysregulated proliferation, invasion and metastasis, angiogenesis, and enhanced cell survival. Therefore, the EGFR-TK is a promising drug target for many types of solid tumors, and its inhibition has potential in both the treatment and prevention of these neoplasias. Based on the structure and function of the EGFR, two antireceptor therapeutic strategies have been developed. The first strategy uses humanized monoclonal antibodies generated against the receptor's ligand-binding, extracellular domain. These antibodies block binding of receptor-activating ligands and, in some cases, can induce receptor endocytosis and downregulation. The second approach uses small molecules that compete with adenosine triphosphate for binding to the receptor's kinase pocket, thus blocking receptor activation and the transduction of postreceptor signals. Early clinical studies suggest that both of these approaches, either alone or in combination with standard anticancer therapies, are well tolerated and can induce clinical responses and tumor stabilization in a variety of common carcinomas. ZD1839 (Iressa; AstraZeneca Pharmaceuticals LP, Wilmington, DE) is the EGFR-TK inhibitor furthest along in clinical development, and it is currently being investigated in a variety of solid tumors, including non-small-cell lung cancer. PMID- 12644980 TI - Studies with ZD1839 in preclinical models. AB - ZD1839 (Iressa; AstraZeneca Pharmaceuticals LP, Wilmington, DE) is an orally active, selective inhibitor of epidermal growth factor receptor-tyrosine kinase (EGFR-TK) that blocks signaling pathways responsible for driving proliferation, invasion, and survival of cancer cells. In preclinical studies of cell lines and human tumor xenografts, ZD1839 as single-agent therapy produced growth inhibition in a wide variety of common solid tumor types including lung, prostate, breast, colon, and ovarian cancers. In these models, ZD1839 inhibited growth of tumor xenografts with high, moderate, and low expression of EGFR. The A431 vulvar carcinoma model, which expresses abnormally high levels of EGFR, was particularly sensitive to ZD1839 treatment, leading to tumor regression. When ZD1839 was coadministered with cytotoxic chemotherapy agents or radiotherapy, additive or even synergistic antitumor activity was achieved. The inhibition observed with ZD1839 treatment was not restricted to advanced metastatic tumors, but also extended to early lesions such as breast xenografts of human ductal carcinoma in situ. Inhibition of EGFR-TK has also been shown to delay the onset of tumor development in a transgenic animal model. The diverse and profound antitumor activities attained with ZD1839 treatment in tumor cells and in xenograft tumor models provided the rationale for clinical development of ZD1839. Ongoing preclinical studies continue to support the importance of EGFR-TK activity in the biology of solid tumors. PMID- 12644982 TI - Dose-comparative monotherapy trials of ZD1839 in previously treated non-small cell lung cancer patients. AB - Patients with non-small cell lung cancer (NSCLC) frequently present, or relapse, with unresectable disease that is resistant to standard chemotherapy. There is, therefore, an urgent need for new treatments for NSCLC and other solid tumors. ZD1839 (Iressa; AstraZeneca Pharmaceuticals LP, Wilmington, DE), an orally active, selective epidermal growth factor receptor-tyrosine kinase inhibitor, has shown promising antitumor responses in phase I clinical trials in heavily pretreated patients with advanced NSCLC and other solid tumors. Randomized, multicenter global and US-based clinical trials were conducted to investigate two doses of ZD1839 as second- or third-line monotherapy in patients with advanced NSCLC. The global trial, Iressa Dose Evaluation in Advanced Lung Cancer (IDEAL) 1, was a double-blind, randomized, dose-comparative study that enrolled 210 patients with NSCLC at centers in Europe, Japan, South Africa, and Australia. This trial included patients with advanced unresectable stage III or IV NSCLC who had recurrent or progressive disease following one or two chemotherapy regimens, at least one of which was platinum based. IDEAL-1 showed that once-daily oral treatment with ZD1839 at 250 or 500 mg/day resulted in tumor response rates of 18% and 19%, respectively. Disease control rates, which included both tumor responses and stable disease, were 54% and 51%, respectively. Median progression free survival was 83 days in the 250 mg/day group and 85 days in the 500 mg/day group. Rapid improvements in NSCLC-related symptoms were seen in the subpopulation of patients who were symptomatic and had completed a Lung Cancer Subscale questionnaire at baseline. Both the 250 mg/day and 500 mg/day doses of ZD1839 were well tolerated by patients in this trial. The majority of adverse events were grades 1 or 2 skin rash and diarrhea, which were readily manageable and reversible, and withdrawals were rare. The US monotherapy study in NSCLC, IDEAL-2, comprised 30 trial centers and enrolled 221 patients with NSCLC for third-line or greater therapy; 216 patients received treatment and were evaluable. This trial included patients with advanced stage III or IV NSCLC who had received two or more chemotherapy regimens that contained platinum and docetaxel, either concurrently or in separate regimens, with most patients having received more than two prior regimens. Although the IDEAL-1 and IDEAL-2 trials were similar in study design, patients in IDEAL-2 were sicker, as evidenced by a higher percentage of patients with a performance status of 2, metastatic disease, and disease-related symptoms. Because measuring the symptom improvement rate was a primary objective in IDEAL-2, all patients were symptomatic and were required to have a Lung Cancer Subscale score of 24 or less at trial entry. Objective tumor response rates (all partial responses) were 12% for the 250 mg/day group and 9% for the 500 mg/day group. Symptom improvement rates (increase of at least two points on the Lung Cancer Subscale) were 43% and 35%, respectively. Both doses of ZD1839 were well tolerated in this trial. The results of IDEAL-1 and IDEAL-2 indicate that ZD1839 monotherapy may offer a single-agent alternative for patients with advanced solid tumors who have received and progressed on prior chemotherapy, many of whom have exhausted their therapy options. PMID- 12644981 TI - Phase I studies of ZD1839 in patients with common solid tumors. AB - Clinical development of ZD1839 (Iressa; AstraZeneca Pharmaceuticals LP, Wilmington, DE) was initiated based on strong preclinical studies that showed antitumor responses in a variety of solid tumor types and established its oral bioavailability and tolerability. In phase I trials, ZD1839 was investigated in patients with a wide variety of solid tumors that commonly express epidermal growth factor receptor-tyrosine kinase. More than 250 patients with advanced refractory disease were enrolled in phase I studies; of these, 100 had non-small cell lung cancer (NSCLC). All patients had been recipients of substantial prior therapy. Intermittent and continuous dosing schedules with ZD1839 were well tolerated by these patients for up to 6 months or more. The most frequently reported adverse events were grade 1 or 2 diarrhea and grade 1 or 2 acneiform rash; these effects were reversible on discontinuation of therapy and either required no management or were easily managed. Grade 3 or 4 adverse events were rare and most were attributed to disease progression rather than being drug related. In phase I trials, the maximum tolerated dose of ZD1839 was determined to be 700 to 1000 mg/day; diarrhea was found to be the dose-limiting toxicity. Pharmacokinetic profiles supported once-daily oral dosing and indicated that biologically active plasma concentrations were achieved at the doses studied. Pharmacodynamic results in skin biopsies showed that ZD1839 produced inhibition of epidermal growth factor receptor-tyrosine kinase in these patients. Durable tumor responses and stable disease were observed with ZD1839 treatment in patients with advanced NSCLC. Symptoms related to NSCLC also appeared to improve rapidly in some patients. Prolonged time on therapy was observed for other tumor types in addition to NSCLC, including hormone-refractory prostate, colorectal, head and neck, breast, ovarian, and renal cancers. The promising phase I results observed in heavily pretreated patients with advanced NSCLC and other solid tumors have warranted investigation in phase II/III clinical trials in these populations. Phase II/III trials are being conducted in patients with advanced NSCLC. Phase II trials of ZD1839 are also being conducted in patients who have hormone-refractory prostate, breast, or colorectal cancer, as well as other solid tumors. Based on the phase I study results, once-daily oral doses of 250 mg and 500 mg, which are well below the maximum tolerated dose, were selected for further clinical investigation of ZD1839. PMID- 12644983 TI - Impact of ZD1839 on non-small cell lung cancer-related symptoms as measured by the functional assessment of cancer therapy-lung scale. AB - The majority of patients with non-small cell lung cancer (NSCLC) present with advanced disease, which is associated with a poor prognosis and symptoms such as pain, coughing, and shortness of breath. In patients who present at an earlier stage, the progressive nature of NSCLC and its resistance to treatment often result in recurrence, with the associated symptoms of advanced disease. These symptoms negatively affect patient quality of life and performance status rating, both of which are predictive of treatment response and survival. There is increasing interest in using assessments of improvements in symptoms and quality of life as outcomes in clinical trials for patients with advanced NSCLC. Patients with NSCLC have limited therapeutic options. Even those patients who are able to tolerate chemotherapy can expect median survival increases of only 2 to 4 months. The new targeted therapies for lung cancer, in contrast, are relatively nontoxic and may provide benefits for symptoms and quality of life in addition to tumor responses. The Functional Assessment of Cancer Therapy-Lung (FACT-L) scale is a validated, sensitive, and reliable patient questionnaire that evaluates and quantifies quality of life across several dimensions, including lung cancer related symptoms (Lung Cancer Subscale). The Lung Cancer Subscale ranges from 0 (severe debilitation) to 28 (asymptomatic). A change of two points reflects a clinically significant change in NSCLC-related symptoms and quality of life. In phase I studies and also in the Iressa Dose Evaluation in Advanced Lung Cancer (IDEAL)-1 and IDEAL-2 phase II monotherapy trials, treatment of patients with advanced NSCLC with the epidermal growth factor receptor-tyrosine kinase inhibitor ZD1839 (Iressa; AstraZeneca Pharmaceuticals LP, Wilmington, DE) has shown tumor responses as well as rapid improvements in NSCLC-related symptoms and quality of life. In IDEAL-1 and IDEAL-2, improvements in NSCLC-related symptoms and quality of life, as measured by FACT-L, correlated with tumor response, and improvements in symptoms also correlated with progression-free and overall survival. Although symptom response is correlated with tumor response, it is also uniquely predictive of progression-free and overall survival. The FACT-L questionnaire has also been included in phase III trials of ZD1839 treatment in combination with chemotherapy regimens. PMID- 12644984 TI - New directions for ZD1839 in the treatment of solid tumors. AB - Among molecular mechanisms of tumor growth and progression, activated epidermal growth factor receptor-tyrosine kinase is remarkable for its prevalence and impact on many different solid tumor types, as well as its link to diverse stages of disease. ZD1839, (Iressa; AstraZeneca Pharmaceuticals LP, Wilmington, DE) an oral epidermal growth factor receptor-tyrosine kinase inhibitor, is also being explored for its potential in the treatment of other common solid tumors, including breast cancer, colorectal cancer, and hormone-refractory prostate cancer. Clinical trials include studies of ZD1839 as monotherapy and in combination with various chemotherapy, radiation, and hormone therapy regimens. ZD1839 is also being studied in conjunction with other targeted therapies, such as trastuzumab. In addition to advanced and refractory disease, ZD1839 may also be applicable for use in earlier stage cancers. Chemoprevention trials have been initiated to assess the potential of ZD1839 to delay or prevent the progression of common solid tumors such as non-small cell lung cancer. PMID- 12644985 TI - Charge state dependent collision-induced dissociation of native and reduced porcine elastase. AB - The [M + 20H](20+)-[M + 12H](12+) charge states of native and reduced porcine elastase, a 25.9 kDa serine protease, were subjected to collisional activation in a quadrupole ion trap. For most charge states, ion parking was used to increase the number of parent ions over that yielded directly by electrospray. Ion-ion proton transfer reactions were used to reduce product ion charge states largely to +1 to simplify spectral interpretation. Both forms of the protein show charge state dependent fragmentation behavior. The native protein, which contains four disulfide linkages, shows almost no evidence for fragmentation within the regions of the protein linked by disulfide bonds. However, at the lowest charge states studied, evidence for cleavage of a least one of the disulfide bonds was evident in the appearance of a c-type ion. The highest charge states of native elastase showed several prominent cleavages C-terminal to valine residues. As the charge state decreased, however, preferential cleavages at acidic amino acid residues became important. The reduced form of the protein did not show particularly prominent cleavages at valine residues. However, many of the same preferential cleavages at acidic amino acid residues noted for the native protein were also observed in the same charge states of the reduced protein. The reduced protein also showed additional cleavages from regions of the protein that are ordinarily protected by disulfide linkages in the native form. PMID- 12644986 TI - Detection of tyrosine phosphorylated peptides via skimmer collision-induced dissociation/ion trap mass spectrometry. AB - Phosphorylation of proteins is an important post-translational protein modification in cellular response to environmental change and occurs in both prokaryotes and eukaryotes. Identification of the amino acid on individual proteins that become phosphorylated in response to extracellular stimulus is essential for understanding the mechanisms involved in the intracellular signals that these modifications facilitate. Most protein kinases catalyze the phosphorylation of proteins on serine, threonine or tyrosine. Although tyrosine phosphorylation is often the least abundant of the three major phosphorylation sites, it is important owing to its role in signal pathways. Currently available methods for the identification of phosphorylation sites can often miss low levels of tyrosine phosphorylations. This paper describes a method for the identification of phosphotyrosine-containing peptides using electrospray ionization on an ion trap mass spectrometer. Skimmer-activated collision-induced dissociation (CID) was used to generate the phosphotyrosine immonium ion at m/z 216. This method is gentle enough that the protonated molecule of the intact peptide is still observed. In-trap CID was employed for the verification of the phosphotyrosine immonium ion. Using this technique, low levels of phosphotyrosine containing peptides can be identified from peptide mixtures separated by nanoflow micro liquid chromatography/mass spectrometry. PMID- 12644987 TI - Gas-phase theoretical prediction of the metal affinity of copper(I) ion for DNA and RNA bases. AB - The most stable tautomeric forms of free DNA and RNA bases were considered as substrates for the interaction of Cu(+) ion. Several suitable attachment sites were selected that involved mono- and bi-coordination of the cation. B3LYP/6-311 + G(2df,2p) bond energies showed that copper ion has the major affinity for guanine and cytosine bases. The proposed values of Cu(+) ion affinity are 59.9, 60.0, 80.2, 88.0 and 69.0 kcal mol(-1) for uracil, thymine, cytosine, guanine and adenine, respectively. The preference for the mono- or bi-coordination depends on the particular tautomer for each base. PMID- 12644988 TI - Improving hydrogen/deuterium exchange mass spectrometry by reduction of the back exchange effect. AB - The measurement of deuterium incorporation kinetics using hydrogen/deuterium (H/D) exchange experiments is a valuable tool for the investigation of the conformational dynamics of biomolecules in solution. Experiments consist of two parts when using H/D exchange mass spectrometry to analyse the deuterium incorporation. After deuterium incorporation at high D(2)O concentration, it is necessary to decrease the D(2)O concentration before the mass analysis to avoid deuterium incorporation under artificial conditions of mass spectrometric preparation and measurement. A low D(2)O concentration, however, leads to back exchange of incorporated deuterons during mass analysis. This back-exchange is one of the major problems in H/D exchange mass spectrometry and must be reduced as much as possible. In the past, techniques using electrospray ionization (ESI) had the lowest back-exchange values possible in H/D exchange mass spectrometry. Methods for the measurement of H/D exchange by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) that have been developed since 1998 have some significant advantages, but they could not achieve the back exchange minima of ESI methods. Here, we present a protocol for H/D exchange MALDI-MS which allows for greater minimization of back-exchange compared with H/D exchange ESI-MS under similar conditions. PMID- 12644989 TI - Analysis of post-translational modification and characterization of the domain structure of dynamin A from Dictyostelium discoideum. AB - The post-translational modifications of the 96 kDa protein dynamin A from Dictyostelium discoideum were analyzed using Q-TOF mass spectrometry. The accurate molecular mass of the intact protein revealed a covalent modification causing an additional mass of 42 Da. The modification could be identified as N terminal acetylation by tandem mass spectrometry. Extracted ion chromatograms for the a(1) and b(1) ion of the tryptic T1 peptide were used to detect the acetylated peptide within 54 nanoelectrospray ionization tandem mass spectra. Owing to the accurate molecular mass of the intact protein, additional covalent modifications could be excluded. In addition to the covalent modification, the domain structure of dynamin A was determined by applying a combination of limited proteolysis, sodium dodecylsulfate polyacrylamide gel electrophoresis, automated tandem mass spectrometry and protein database searching. PMID- 12644990 TI - Screening, library-assisted identification and validated quantification of fifteen neuroleptics and three of their metabolites in plasma by liquid chromatography/mass spectrometry with atmospheric pressure chemical ionization. AB - A liquid chromatographic/mass spectrometric assay with atmospheric pressure chemical ionization (APCI-LC/MS) is presented for the fast and reliable screening and identification and for the precise and sensitive quantification of 15 neuroleptic (antipsychotic) drugs and three of their relevant metabolites in plasma. It allows confirmation of the diagnosis of a neuroleptic overdose and monitoring of psychiatric patients' compliance. The neuroleptics amisulpride, bromperidol, clozapine, droperidol, flupenthixol, fluphenazine, haloperidol, melperone, olanzapine, perazine, pimozide, risperidone, sulpiride, zotepine and zuclopenthixol and the pharmacologically active metabolites norclozapine, clozapine N-oxide and 9-hydroxyrisperidone were extracted from plasma using solid phase extraction and were separated on a Merck LiChroCART column with Superspher 60 RP Select B as the stationary phase. Gradient elution was performed using aqueous ammonium formate and acetonitrile. After screening and identification in the scan mode using the authors' new LC/MS library, the neuroleptics were quantified in the selected-ion mode. The quantification assay was fully validated. It was found to be selective and proved to be linear from sub therapeutic to over therapeutic concentrations for all analytes. The corresponding reference levels are listed. The accuracy and precision data were within the required limits. The analytes were stable in frozen plasma for at least 1 month. The method was successfully applied to several authentic plasma samples from patients treated or intoxicated with various neuroleptics. The validated LC/MS assay has proved to be appropriate for the isolation, separation, screening, identification and quantification of various neuroleptics in plasma for clinical toxicology and therapeutic drug monitoring purposes. PMID- 12644992 TI - Reactions of gaseous acylium ions with 1,3-dienes: further evidence for polar [4 + 2+] Diels-Alder cycloaddition. AB - A novel reaction of acylium and thioacylium ions, polar [4 + 2(+)] Diels-Alder cycloaddition with 1,3-dienes and O-heterodienes, has been systematically investigated in the gas phase (Eberlin MN, Cooks RG. J. Am. Chem. Soc. 1993; 115: 9226). This polar cycloaddition, yet without precedent in solution, likely forms cyclic 2,5-dihydropyrylium ions. Here we report the reactions of gaseous acylium ions [(CH(3))(2)N-C(+)=O, Ph-C(+)=O, (CH(3))(2)N-C(+)=S, CH(3)-C(+)=O, CH(3)CH(2) C(+)=O, and CH(2)=CH-C(+)=O] with several 1-oxy-substituted 1,3-dienes of the general formula RO-CH=CH-C(R(1))=CH(2), which were performed to collect further evidence for cycloaddition. In reactions with 1-methoxy and 1-(trimethylsilyloxy) 1,3-butadiene, adducts are formed to a great extent, but upon collision activation they mainly undergo structurally unspecific retro-addition dissociation. In reactions with Danishefsky's diene (trans-1-methoxy-3 (trimethylsilyloxy)-1,3-butadiene), adducts are also formed to great extents, but retro-addition is no longer their major dissociation; the ions dissociate instead mainly to a common fragment, the methoxyacryl cation of m/z 85. This fragment ion is most likely formed with the intermediacy of the acyclic adduct, which isomerizes prior to dissociation by a trimethylsilyl cation shift. Theoretical calculations predict that meta cycloadducts bearing 1-methoxy and 1 trimethylsilyloxy substituents are unstable, undergoing barrierless ring opening induced by the charge-stabilizing effect of the 1-oxy substituents. In contrast, for the reactions with 1-acetoxy-1,3-butadiene, both the experimental results and theoretical calculations point to the formation of intrinsically stable cycloadducts, but the intact cycloadducts are either not observed or observed in low abundances. Both the isomeric ortho and meta cycloadducts are likely formed, but the nascent ions dissociate to great extents owing to excess internal energy. The ortho cycloadducts dissociate by ketene loss; the meta cycloadducts undergo intramolecular proton transfer to the acetoxy group followed by dissociation by acetic acid loss to yield aromatic pyrylium ions. Either or both of these dissociations, ketene and/or acetic acid loss, dominate over the otherwise favored retro-Diels-Alder alternative. The pyrylium ion products therefore constitute compelling evidence for polar [4 + 2(+)] cycloaddition since their formation can only be rationalized with the intermediacy of cyclic adducts. PMID- 12644991 TI - A miniaturized matrix-assisted laser desorption/ionization time of flight mass spectrometer with mass-correlated acceleration focusing. AB - Mass correlated acceleration (MCA) has now been integrated into a 4 in (10.2 cm) matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometer to achieve high resolving power across a broader mass range, without sacrificing detection of higher mass ions. The goal was to combine MCA with a custom-built miniaturized instrument such as those that might be used for field portable applications. Unlike other pulsed extraction methods, MCA is not mass dependent and mass spectra can be achieved with a single tuning of instrument parameters. Additionally, the multi-channel recording advantage is better realized because ions of all masses can be brought into focus simultaneously. The MCA dual-stage ion source compensates dynamically for the mass dependence by incorporating an extraction pulse region followed by an acceleration region that contains a time-dependent waveform correlated with mass. The technique was validated with applications in peptide mixtures and protein digestions. Diagnostic studies for the instrument include m/z range and limits of detection. PMID- 12644993 TI - Macromolecular assembly of Helicobacter pylori urease investigated by mass spectrometry. AB - The supramolecular assembly of Helicobacter pylori urease was studied by nanoflow electrospray ionization orthogonal time-of-flight mass spectrometry. The measured molecular mass of the urease complex of 1.06 MDa corresponds to a dodecameric (alphabeta)(12) assembly of urease alpha (26 kDa) and beta (61 kDa) subunits. The dodecamer disassembles readily into (alphabeta)(3) subunits in solution and under controlled collisional-induced dissociation in the gas phase. This is in strong support of an ((alphabeta)(3))(4) architecture consistent with the recently published x-ray structure. In vitro, the alpha and beta subunits are capable of re-assembling to (alphabeta)(3), but not further to the dodecameric complex. PMID- 12644994 TI - Complexes of cadmium ion with guanine bases detected by electrospray ionization mass spectrometry. AB - The complexations of cadmium ion with guanine bases were detected by electrospray ionization mass spectrometry (ESI-MS). In order to explore the toxicity of cadmium, such as oxidative stress to DNA and carcinogenesis, it is very important to determine the interactions between the cadmium ion and nucleotide. The analysis of mixed cadmium ion-guanosine aqueous solution (molar ratio 1 : 9) using ESI-MS (cone voltage 20 V) showed the presence of various cadmium complex ions, such as [n (guanosine) + Cd](2+) (n = 3-8), [2guanine + Cd](2+), [guanosine + guanine + Cd](2+) and [guanosine + 2guanine + Cd](2+). The observed [2guanine + Cd](2+), [guanosine + guanine + Cd](2+) and [guanosine + guanine + Cd](2+) ions are formed through the dissociation of the N-glycoside bond at the interface of ESI-MS. For deoxyguanosine and ethylguanine, similar cadmium complexes were observed. However, the complexes between the cadmium ion and 8 hydroxydeoxyguanosine were not detected. Furthermore, when a higher molar ratio (Cd : guanosine) or cone voltage were used, more of the monovalent ion peaks, such as [Cd(guanine - H)(2) + H](+) and [Cd(guanosine - H)(2) + H](+), were observed and a decrease in the abundance of the divalent ions, such as [n(guanosine)+Cd](2+), occurred. PMID- 12644996 TI - Tandem mass spectrometry of Cu(II) complexes: the effects of ligand donor group on dissociation. AB - A quadrupole ion trap mass spectrometer was used to study the dissociation patterns of Cu(II) complexes with various linear podand ligands. Cu(II) complexes having different combinations of nitrogen-, oxygen- and sulfur-containing terminal functionality attached to a diethylenetriamine (DIEN) framework were ionized by electrospray and collision-induced dissociation (CID) was used to generate product ions. Regardless of the particular functional groups present, the complexes undergo predominantly heterolytic cleavages of carbon-carbon bonds along the DIEN backbone with Cu remaining coordinated to one of the two terminal functional groups. Upon dissociation, Cu's preference to remain coordinated to a particular functional group follows the trend thioether > amine > imidazole > pyridine > ether. A simple evaluation of this trend based upon metal-functional group binding affinity appears not to be adequate for fully explaining these observations. The tendency of Cu(II) to be reduced upon dissociation helps explain the observed trend, as does the flexibility of the functional group, which affects its ability to orient its dipole effectively toward the metal. PMID- 12644995 TI - Characterization of geometrid sex pheromones by electrospray ionization time-of flight mass spectrometry. AB - The utility of liquid chromatography combined with time-of-flight mass spectrometry (LC/TOFMS) was demonstrated for studies on chiral unsaturated epoxy compounds, sex pheromones produced mainly by female moths in the family Geometridae. By electrospray ionization (ESI), each synthetic epoxyalkadiene derived from (Z,Z,Z)-3,6,9-triene with a C(18)-C(23) straight chain showed three ion series, [M + NH(4)](+), [M + H](+) and [M - OH](+), with high resolution and good sensitivity, indicating its molecular formula. In addition to these, characteristic fragment ions at m/z M - 57 and M - 71 for the 3,4-epoxides and at m/z M - 123 and 123 for the 9,10-epoxides were detected, whereas the 6,7-epoxides did not produce fragment ions that reflected their structures. Monitoring these diagnostic ions during the LC/MS analysis of a gland extract, the natural sex pheromone of the mulberry looper was confirmed to be (Z,Z)-cis-9,10-epoxy-3,6 octadecadiene, which was separable from the other positional isomers on an ODS column. Furthermore, (Z,Z)-cis-3,4-epoxy-6,9-nonadecadiene secreted by the Japanese giant looper was analyzed with a chiral column, and the stereochemistry was determined directly. PMID- 12644997 TI - Differentiation between isomeric 2-thionosparteine and 15-thionosparteine on the basis of electron ionization mass spectra. PMID- 12644999 TI - Current literature in journal of mass spectrometry. PMID- 12645002 TI - Intracellular and extracellular roles of S100 proteins. AB - S100, a multigenic family of non-ubiquitous Ca(2+)-modulated proteins of the EF hand type expressed in vertebrates exclusively, has been implicated in intracellular and extracellular regulatory activities. Members of this protein family have been shown to interact with several effector proteins within cells thereby regulating enzyme activities, the dynamics of cytoskeleton constituents, cell growth and differentiation, and Ca(2+) homeostasis. Structural information indicates that most of S100 proteins exist in the form of antiparallelly packed homodimers (in some cases heterodimers), capable of functionally crossbridging two homologous or heterologous target proteins in a Ca(2+)-dependent (and, in some instances, Ca(2+)-independent) manner. In addition, extracellular roles have been described for several S100 members, although secretion (via an unknown mechanism) has been documented for a few of them. Extracellular S100 proteins have been shown to exert regulatory effects on inflammatory cells, neurons, astrocytes, microglia, and endothelial and epithelial cells, and a cell surface receptor, RAGE, has been identified as a potential S100A12 and S100B receptor transducing the effects of these two proteins on inflammatory cells and neurons. Other cell surface molecules with ability to interact with S100 members have been identified, suggesting that RAGE might not be a universal S100 protein receptor and/or that a single S100 protein might interact with more than one receptor. Collectively, these data indicate that members of the S100 protein family are multifunctional proteins implicated in the regulation of a variety of cellular activities. PMID- 12645003 TI - Molecular mechanisms of S100-target protein interactions. AB - S100 proteins have no known enzymatic activity and exert their intracellular effects via interaction with and regulation of the activity of other proteins, termed target proteins, in both a Ca(2+)-dependent and Ca(2+)-independent manner. Structural studies have identified the linker region between the two EF-hand Ca(2+) binding domains and the C-terminus as Ca(2+)-dependent target protein binding sites in several S100 family members. In fact, C-terminal aromatic residues are obligatory for interaction of S100A1 with several of its Ca(2+) dependent target proteins. Pharmacological studies suggest the presence of additional Ca(2+)-dependent binding motifs on some family members. A minimum of seven family members interact with and regulate the activity of aldolase A in a Ca(2+)-independent manner. In the case of S100A1, Ca(2+)-independent target protein interactions utilize a binding motif distinct from the C-terminal Ca(2+) dependent target protein binding site. Several studies suggest that ionic interactions participate in the interaction of S100 family members with Ca(2+) independent target proteins. While some target proteins are activated by multiple family members, other target proteins exhibit family member-specific activation, i.e., they are activated by a single family member. As predicted, family member specific interactions appear to be mediated by regions that exhibit the most divergence in amino acid sequence among family members, the linker or "hinge" region and the C terminus. Further specificity in S100-target protein interactions may arise from the different biochemical/biophysical properties of the individual family members, including affinity for metal ions (Ca(2+), Zn(2+), and Cu(2+)), oligomerization properties, heterodimerization, post-translational modifications, and lipid-binding. Delineation of the structural motifs that mediate S100-target protein interactions and determination of the in vivo relevance of these interactions are needed to fully understand the role of S100 proteins in normal and diseased cells. PMID- 12645004 TI - Monitoring of S100 homodimerization and heterodimeric interactions by the yeast two-hybrid system. AB - The S100 family consists of 19 members, which function as transducers of calcium signals in a tissue-specific manner. Upon calcium binding, the conformation of many S100 proteins changes dramatically. Several hydrophobic residues are exposed, allowing the S100 proteins to interact with their target proteins, and thereby to transduce calcium signals into specific biological responses. To further elucidate the exact contribution of the S100 calciproteins in the calcium signalling pathways, several groups have applied the yeast two-hybrid technology to identify putative target proteins for the various S100 calciproteins. Two hybrid large screens using S100 proteins as baits have confirmed the biochemical and structural feature of S100, which enable them to form homodimers and the ability of some members to form specific heterodimers in vivo. Yeast two-hybrid investigations have allowed the identification of conserved hydrophobic residues and domains that are crucial for the stabilization of S100 homo- and heterodimers. Furthermore, this method clearly underlines that the homo- and heterodimerization mechanisms differ among the members of the S100 family. However, several lines of evidence strongly suggest that two-hybrid methodology is limited to the analysis of interactions that are calcium-independent, since no target proteins other than S100 family members themselves have been detected with this methodology. PMID- 12645005 TI - S100A9/S100A8: Myeloid representatives of the S100 protein family as prominent players in innate immunity. AB - Neutrophils are rapidly recruited to sites of inflammation and are thereby at the forefront of the organism's defense against numerous attacks. As unspecific phagocytes, they belong to the so-called innate immunity. Two S100 proteins, namely S100A9 (MRP14) and S100A8 (MRP8), constitute roughly 40% of the cytosolic protein in these cells, implying by their pure abundance an important role in the effector functions of neutrophils. However, despite intense research in the past 15 years, the puzzle that may embed both molecules into the neutrophil/monocyte physiology is still incomplete. One reason might be the conformational variability the S100A9 and S100A8 molecules can adopt. They readily form hetero- and homodimeric, trimeric as well as tetrameric complexes, but they evidently do also exert specific functions as monomers. An ever-increasing body of information suggests that S100A9 plays a prominent role in leukocyte trafficking and arachidonic acid metabolism. In addition, elevated levels of S100A9 and S100A8 in body fluids of inflamed tissues strengthen the view that these molecules are important players in fighting inflammation. The aim of this review is to give an update on the current developments concerning the S100A9/S100A8 molecule in biology and medicine. PMID- 12645006 TI - Multiple structural states of S100A12: A key to its functional diversity. AB - S100A12 is a member of the S100 family of EF-hand calcium-binding proteins. Together with two other calgranulins, S100A8 and S100A9, it is mostly expressed in human granulocytes, although there is increasing evidence of expression in keratinocytes and psoriatic lesions. It is involved in host-parasite response, and linked to corneal autoimmune diseases connected with filarial parasite infestation. Interaction of S100A12 with a multiligand receptor for advanced glycation end products (RAGE) mediates inflammation. Human recombinant S100A12 was found to induce neuritogenesis of cultured hippocampal cells, similar to two other S100 proteins, S100B and S100A4. X-ray structure of S100A12 has been solved in two crystal forms: R3 and P2(1). In the R3 crystal form S100A12 is a dimer, and in the P2(1) crystal form the dimers are arranged as a hexamer. The hexameric form suggests its role in receptor oligomerisation. S100A12 binds copper at the predicted zinc/copper binding site, which is located close to the surface of the protein. We propose copper-mediated generation of reactive oxygen species by S100A12 as its function in host-parasite response. PMID- 12645007 TI - Frog p26olf, a molecule with two S100-like regions in a single peptide. AB - An S100-like calcium-binding protein, p26olf, was originally isolated from the frog (Rana Catesbeiana) olfactory epithelium with four chromatographical steps. The primary structure of p26olf contains two S100-like regions aligned in tandem with four functional EF-hands. At 100 mM K(+), wild-type p26olf binds Ca(2+) with a Kd value of 22 microM and a Hill coefficient of 2.0. Each EF-hand seems to have different affinity for Ca(2+): it is high in EF-A and -B and low in EF-C and -D. In our Ca(2+)-binding model, the order of Ca(2+)-binding to p26olf is EF-B, EF-A, EF-C, and EF-D. Expression of mRNA of p26olf is detected in various frog tissues: it is high in the olfactory epithelium, lung, and spleen, moderate in brain, retina, heart, and kidney, and low in liver and muscle. Immunohistochemical studies revealed that p26olf is prominently localized in the cilia of both olfactory and lung respiratory epithelium and especially enriched in the distal segment of the olfactory cilia. Several proteins in the olfactory cilia bind to p26olf in the presence of Ca(2+), suggesting that they are possible target proteins of p26olf. One of these target proteins is immunologically identified as a beta-adrenergic receptor kinase-like protein. In the olfactory cilia, p26olf may have some roles in the olfactory transduction or adaptation through interaction with this beta-adrenergic receptor kinase-like protein. PMID- 12645008 TI - Differential expression of S100 proteins in the developing human hippocampus and temporal cortex. AB - S100 calcium binding proteins have long been known to express in the adult nervous system, but their distribution in the developing brain, especially the human fetal brain, is largely unknown. We used an immunohistochemical method to determine the expression of three S100 proteins, namely S100A4, S100A5, and S100A13, in the human fetal hippocampus and temporal cortex from 12 to 33 weeks of gestation. At 12 weeks, S100A5 was strongly expressed in the cells and fibers of the polymorphic, pyramidal, and molecular layers of the hippocampus. Thereafter, its expression decreased with age. In the temporal cortex, S100A5 expression was detected from 12 weeks onwards, peaked at 20 to 24 weeks, and then decreased with age. The horizontal fibers of the marginal zone were immunoreactive at all stages examined. S100A13 immunoreactivity was also detected in both cells and fibers of the hippocampus at 12 weeks, became slightly stronger at 20 weeks, and then decreased with age. In the temporal cortex, S100A13 immunoreactivity was also strong in all cellular layers at 12 to 24 weeks before it declined with age from 28 weeks onwards. Among the three proteins examined, S100A4 showed the weakest expression, which was detected in the cells and fibers of the hippocampus and the temporal cortex at all stages examined. Our results have demonstrated for the first time, in the human fetal hippocampus and temporal cortex, specific spatio-temporal patterns of expression of these proteins, all of which are likely to have different roles to play during development despite their pronounced sequence homology. PMID- 12645009 TI - S100B in brain damage and neurodegeneration. AB - S100B is a calcium-binding peptide produced mainly by astrocytes that exert paracrine and autocrine effects on neurons and glia. Some knowledge has been acquired from in vitro and in vivo animal experiments to understand S100B's roles in cellular energy metabolism, cytoskeleton modification, cell proliferation, and differentiation. Also, insights have been gained regarding the interaction between S100B and the cerebral immune system, and the regulation of S100B activity through serotonergic transmission. Secreted glial S100B exerts trophic or toxic effects depending on its concentration. At nanomolar concentrations, S100B stimulates neurite outgrowth and enhances survival of neurons during development. In contrast, micromolar levels of extracellular S100B in vitro stimulate the expression of proinflammatory cytokines and induce apoptosis. In animal studies, changes in the cerebral concentration of S100B cause behavioral disturbances and cognitive deficits. In humans, increased S100B has been detected with various clinical conditions. Brain trauma and ischemia is associated with increased S100B concentrations, probably due to the destruction of astrocytes. In neurodegenerative, inflammatory and psychiatric diseases, increased S100B levels may be caused by secreted S100B or release from damaged astrocytes. This review summarizes published findings on S100B regarding human brain damage and neurodegeneration. Findings from in vitro and in vivo animal experiments relevant for human neurodegenerative diseases and brain damage are reviewed together with the results of studies on traumatic, ischemic, and inflammatory brain damage as well as neurodegenerative and psychiatric disorders. Methodological problems are discussed and perspectives for future research are outlined. PMID- 12645010 TI - S-100 proteins in the human peripheral nervous system. AB - This article reviews the distribution of S100 proteins in the human peripheral nervous system. The expression of S100 by peripheral glial cells seems to be a distinctive fact of these cells, independently of their localization and their ability to myelinate or not. S100 proteins expressing cells include satellite cells of sensory, sympathetic and enteric ganglia, supporting cells of the adrenal medulla, myelinating and non-myelinating Schwann cells in the nerve trunks, and the Schwann-related cells of sensory corpuscles. In addition, S100 proteins are expressed in peripheral neurons. Most of them express S100alpha protein, and a subpopulation of sensory neurons in dorsal root ganglia contains S100beta protein or S100alpha plus S100beta proteins. PMID- 12645011 TI - Subcellular localization of S100A11 (S100C) in LLC-PK1 renal cells: Calcium- and protein kinase c-dependent association of S100A11 with S100B and vimentin intermediate filaments. AB - The subcellular localization of the Ca(2+)-modulated protein, S100A11, was investigated in the renal cell line LLC-PK1 by immunofluorescence and confocal laser scanning microscopy under varying experimental conditions. In control cells, S100A11 was detected on the plasma membrane, where the protein co localized with annexin I (ANXA1) at discrete sites, and found diffusely in the cytoplasm. Elevation of the cytosolic Ca(2+) concentration by means of the Ca(2+) ionophore, ionomycin, caused a significant fraction of S100A11 to associate with vimentin intermediate filament (IF)-bound S100B, another member of the S100 protein family. Under these conditions, ANXA1 underwent a quite different kind of relocation. Translocation of S100A11 onto vimentin IF-bound S100B was also observed upon activation of protein kinase C (PKC). Under these conditions, S100A11 appeared to associate directly with vimentin IFs at cell sites displaying low or no abundance of S100B such as cell processes, and, again, S100A11 and ANXA1 underwent a different relocation. Our data suggest the possibility that the intracellular Ca(2+) level might regulate the subcellular localization of S100A11 and its interaction with definite target proteins, and that S100A11 might serve the function of modulating S100B activities. Interestingly, in spite of the known ability of S100A11 to form heterotetramers with ANXA1, the two proteins underwent a different relocation on elevation of the cytosolic Ca(2+) concentration or activation of PKC, pointing to different regulatory activities of individual proteins in renal cells. PMID- 12645012 TI - Immunohistochemical localization of S100-like protein in non-mammalian kidney. AB - The immunolocalization of S100-like protein was investigated in the kidney of saltwater fishes (Dicentrarchus labrax; Coris julis; Serranus cabrilla; Scorpaena porcus), amphibia (Rana aesculenta), reptiles (Lacerta viridis), and aves (Gallus domesticus; Strutio camelus). S100-like immunoreactivity was detected in the juxtaglomerular cells of all saltwater fishes studied. No immunoreactivity was observed in other tracts of the nephron or in the interstitial tissue. In frog kidney, S100-like immunoreactive cells were localized in the proximal tubule, singly distributed or placed side by side in clusters of two or three cells. S100 like immunoreactive cells were distributed in the distal and in the collecting tubules in lizard, chicken, and ostrich kidney. In the distal tubule of lizard kidney, S100-like immunoreactive cells were numerous and uniformly distributed. In lizard collecting tubules, S100-like immunoreactive cells showed less intense immunoreactivity than in the distal tubule, except for a cluster of cells at the junction with the initial collecting duct. In chicken and ostrich kidney, S100 like immunoreactive cells of the distal tubules were closely packed together. In the collecting tubules, S100-like immunoreactive cells were alternate to negative cells. These results indicate the high conservation degree of S100 proteins through phylogenesis and suggest a functional role for these proteins in the vertebrate kidney. PMID- 12645014 TI - Non-metathetic behavior patterns of Grubbs' carbene. AB - Grubbs' carbene complex [Cl(2)(Cy(3)P)(2)Ru=CHPh] and its second-generation counterpart [Cl(2)(Im)(Cy(3)P)Ru=CHPh] have demonstrated remarkable efficiency metathesizing olefins. Furthermore, a growing number of newly discovered catalytic processes mediated by this complex broaden its synthetic utility beyond olefin metathesis. In addition, the ready availability of these stable ruthenium based catalysts coupled with their tolerance toward a wide variety of common functional groups make Grubbs' catalysts very convenient synthetic tools. This paper presents recent advances in the non-metathetic behavior patterns of Grubbs' carbene. PMID- 12645015 TI - Structural, photophysical, and electrophosphorescent properties of platinum(II) complexes supported by tetradentate N2O2 chelates. AB - We present an examination of the structural and photophysical characteristics of [Pt(N(2)O(2))] complexes bearing bis(phenoxy)diimine auxiliaries (diimine=4,7 Ph(2)phen (1) and 4,4'-tBu(2)bpy (2)) that are tetradentate relatives of the quinolinolato (q) ligand. These neutral derivatives display high thermal stability (>400 degrees C in N(2)). While the crystal lattice in 1 consists of (head-to-tail)-interacting dimers, molecules of 2 are arranged into infinitely stacked planar sheets with possible pi-pi interactions but no close Pt.Pt contacts. Complexes 1 and 2 exhibit moderately intense low-energy UV/Vis absorptions around lambda=400-500 nm that undergo negative solvatochromic shifts. Both derivatives are highly luminescent in solution at 298 K with emission lifetimes in the micros range, and mixed (3)[l-->pi*(diimine)] (l=lone pair/phenoxide) and (3)[Pt(d)-->pi*(diimine)] charge-transfer states are tentatively assigned. The excited-state properties of 2 are also investigated by time-resolved absorption spectroscopy and by quenching experiments with pyridinium acceptors to estimate the excited-state redox potential. These emitters have been employed as electrophosphorescent dopants in multilayer OLEDs. Differences between the brightness, color, and overall performance of devices incorporating 1 and 2 are attributed to the influence of the diimine substituents. PMID- 12645016 TI - Multisite catalysis: a mechanistic study of beta-lactone synthesis from epoxides and CO--insights into a difficult case of homogeneous catalysis. AB - Carbonylation of epoxides with a combination of Lewis acids and cobalt carbonyls was studied by both theoretical and experimental methods. Only multisite catalysis opens a low-energy pathway for trans opening of oxirane rings. This ring-opening reaction is not easily achieved with a single-site metal catalyst due to structural and thermodynamic constraints. The overall reaction pathway includes epoxide ring opening, which requires both a Lewis acid and a tetracarbonylcobaltate nucleophile, yielding a cobalt alkyl-alkoxy-Lewis acid moiety. After CO insertion into the Co-C(alkyl) bond, lactone formation results from a nucleophilic attack of the alkoxy Lewis acid entity on the acylium carbon atom. A theoretical study indicates a marked influence of the Lewis acid on both ring-opening and lactone-formation steps, but not on carbonylation. Strong Lewis acids induce fast ring opening, but slow lactone formation, and visa versa: a good balance of Lewis acidity would give the fastest catalytic cycle as all steps have low barriers. Experimentally, carbonylation of propylene oxide to beta butyrolactone was monitored by online ATR-IR techniques with a mixture of tetracarbonylcobaltate and Lewis acids, namely BF(3), Me(3)Al, Et(2)Al(+).diglyme, and a combination of Me(3)Al/dicobaltoctacarbonyl. We found that the last two mixtures are extremely active in lactone formation. PMID- 12645017 TI - Shape and symmetry of heptacoordinate transition-metal complexes: structural trends. AB - The stereochemistries of heptacoordinate transition-metal complexes are analyzed by using continuous symmetry and shape measures of their coordination spheres. The distribution of heptacoordination through the transition-metal series is presented based on structural database searches including organometallic and Werner-type molecular complexes, metalloproteins, and extended solids. The most common polyhedron seems to be the pentagonal bipyramid, while different preferences are found for specific families of compounds, as in the complexes with three or four carbonyl or phosphine ligands, which prefer the capped octahedron or the capped trigonal prism rather than the pentagonal bipyramid. The symmetry maps for heptacoordination are presented and shown to be helpful for detecting stereochemical trends. The maximal symmetry interconversion pathways between the three most common polyhedra are defined in terms of symmetry constants and a large number of experimental structures are seen to fall along those paths. PMID- 12645018 TI - Novel carboranyl C-glycosides for the treatment of cancer by boron neutron capture therapy. AB - The synthesis of the novel unprotected carboranyl C-glycosides 2 and 20-24 starting from ethynyl C-glycosides 1, 5-8, 10, and 13 is described. The new compounds are highly water-soluble and display only a very low cytotoxicity, which makes them promising candidates for use in boron neutron capture therapy for the treatment of cancer. PMID- 12645019 TI - Formation, structure and reactivity of boryloxycarbyne complexes of group 6 metals. AB - Reaction of the diborane(4) B(2)(NMe(2))(2)I(2) with two equivalents of K[(eta(5) C(5)H(5))M(CO)(3)] (M=Cr, Mo, W) yielded the dinuclear boryloxycarbyne complexes [[(eta(5)-C(5)H(5))(OC)(2)M(triple bond)CO](2)B(2)(NMe(2))(2)] (4 a, M=Mo; b, M=W; c, M=Cr), which were fully characterised in solution by multinuclear NMR methods. The Mo and W complexes 4 a, b proved to be kinetically favoured products of this reaction and underwent quantitative rearrangement in solution to afford the complexes [[(eta(5)-C(5)H(5))(OC)(2)M(triple bond)CO]B(NMe(2))B(NMe(2))[M(CO)(3)(eta(5)-C(5)H(5))]] (5 a, M=Mo; b, M=W); 5 a was characterised by X-ray crystallography in the solid state. Corresponding reactions of B(2)(NMe(2))(2)I(2) with only one equivalent of K[(eta(5) C(5)H(5))M(CO)(3)] (M=Mo, W) initially afforded 1:1 mixtures of the boryloxycarbyne complexes 4 a, b and unconsumed B(2)(NMe(2))(2)I(2). This mixture, however, yielded finally the diborane(4)yl complexes [(eta(5) C(5)H(5))(OC)(3)M[B(NMe(2))B(NMe(2))I]] (6 a, M=Mo; b, M=W) by [(eta(5) C(5)H(5))(OC)(3)M] transfer and rearrangement. Density functional calculations were carried out for 4 c and 5 a, b. PMID- 12645020 TI - Morphogenetic control of calcite crystal growth in sulfonic acid based hydrogels. AB - In this paper the mineralization of CaCO(3) in various hydrogel matrices is presented. Sulfonic acid based hydrogels were prepared by introduction of sulfonate-containing monomers into a polyacrylamide network. The sulfonate content of polyacrylamide-co-vinylsulfonate and polyacrylamide-co-allylsulfonate decreases during elution of the copolymers in demineralized water, indicating insufficient linking of the sulfonate-bearing monomers within the hydrogel. In contrast to this, acrylamidomethylpropanesulfonate (AMPS) effectively copolymerizes with acrylamide (AAm) monomers. To study the influence of spatial arrangement of ionic functional groups within hydrogel networks on the mineralization of CaCO(3), AMPS copolymers with different degrees of AMPS cross linking were synthesized. For the mineralization experiments the copolymers were placed into a double-diffusion arrangement. Calcite as the thermodynamically stable modification of CaCO(3) was obtained with a particular morphology. The pseudocubic habitus resembles aggregates obtained by mineralization in pure polyacrylamide. However, closer examination of the aggregates by scanning electron microscopy (SEM) shows that the crystal growth in the AMPS copolymers is different from that observed in polyacrylamide. Whereas the morphology of the calcite aggregates could be fine-tuned by using copolymers with different sulfonate content, the spatial distribution of the ionic functional groups alters the course of crystallization. Calcium ions are locally accumulated due to the heterogeneous distribution of functional sulfonate groups within the copolymer network. Thereby the nucleation of calcite is triggered, resulting in enhanced mineralization. PMID- 12645022 TI - Synthesis, X-ray powder structure, and magnetic properties of layered NiII methylphosphonate, [Ni(CH3PO3)(H2O)], and NiII octadecylphosphonate, [Ni(CH3 (CH2)17-PO3)(H2O)]. AB - [Ni(CH(3)PO(3))(H(2)O)] (1) and [Ni(CH(3)-(CH(2))(17)-PO(3))(H(2)O)] (2) were synthesised by reaction of NiCl(2).6 H(2)O and the relevant phosphonic acid in water in presence of urea. The compounds were characterised by elemental and thermogravimetric analyses, UV-visible and IR spectroscopy, and their magnetic properties were studied by using a SQUID magnetometer. The crystal structure of 1 was determined "ab initio" from X-ray powder diffraction data and refined by the Rietveld method. The crystals of 1 are orthorhombic, space group Pmn2(1), with a=5.587(1), b=8.698(1), c=4.731(1) A. The compound has a hybrid, layered structure made up of alternating inorganic and organic layers along the b direction of the unit-cell. The inorganic layers consist of Ni(II) ions octahedrally coordinated by five phosphonate oxygen atoms and one oxygen atom from the water molecule. These layers are separated by bilayers of methyl groups and van der Waals contacts are established between them. A preliminary structure characterisation of compound 2 suggests the crystallisation in the orthorhombic system with the following unit-cell parameters: a=5.478(7), b=42.31(4), c=4.725(3) A. The oxidation state of the Ni ion in both compounds is +2, and the electronic configuration is d(8) (S=1), as determined from static magnetic susceptibility measurements above 50 K. Compound 1 obeys the Curie-Weiss law at temperatures above 50 K; the Curie (C) and Weiss (theta) constants were found to be 1.15 cm(3) K mol(-1) and -32 K, respectively. The negative value of theta indicates an antiferromagnetic exchange coupling between near-neighbouring Ni(II) ions. No sign of 3D antiferromagnetic long-range order is observed down to T=5 K, the lowest measured temperature. Compound 2 is paramagnetic above T=50 K, and the values of C and theta were found to be 1.25 cm(3) K mol(-1) and -24 K, respectively. Below 50 K the magnetic behavior of 2 is different from that of 1. Zero-field cooled (zfc) and field-cooled (fc) magnetisation plots do not overlap below T=21 K. The irreversible magnetisation, DeltaM(fc-zfc), obtained as a difference from fc and zfc plots starts to increase at T=20 K, on lowering the temperature, and it becomes steady at T=5 K. The presence of spontaneous magnetisation below T=20 K indicates a transition to a weak-ferromagnetic state for compound 2. PMID- 12645021 TI - Characterization of the 310-helix in model peptides by HRMAS NMR spectroscopy. AB - A tetra- and a hepta-homopeptide from the C(alpha)-tetrasubstituted Aib (alpha aminoisobutyric acid) residue were covalently linked to the POEPOP resin by the fragment-condensation approach. The conformational preferences of the two model peptides were determined for the first time on a solid support by means of high resolution magic angle spinning NMR spectroscopy. The results obtained indicate that the Aib homopeptides adopt a regular 3(10)-helical structure even when they are covalently bound to a polymeric matrix, and thus confirm the remarkable conformational stability of the peptides rich in this amino acid. An ATR-FTIR spectroscopic investigation, performed in parallel, also confirmed that these polymer-bound peptides do indeed adopt a helical conformation. The results of this study open the possibility to exploit the peptide-resin conjugates based on C(alpha)-tetrasubstituted alpha-amino acids as helpful, structurally organized templates in molecular recognition studies or as catalysts in asymmetric synthesis. PMID- 12645023 TI - Tetra- and octalactam macrocycles and catenanes with exocyclic metal coordination sites: versatile building blocks for supramolecular chemistry. AB - The synthesis of a new tetralactam macrocycle and the simultaneous formation of catenanes and larger octalactam macrocycles is reported. These species bear 2,2' biquinoline moieties suitably positioned to bind a metal center at the outer periphery of the macrocycles. (1)H NMR chemical shifts permit the unambiguous distinction of transoid and cisoid conformations of the biquinoline moiety, thereby allowing an unequivocal identification of the catenane and octalactam structures, despite the fact that both have the same elemental composition and bear identical structural subunits. With the aid of an anion template effect, rotaxanes can be prepared from the smaller tetralactam macrocycle. These reveal significantly altered requirements in terms of the stopper size as compared to previously reported tetralactam wheels. Several copper(I)-mediated dimers and a (bpy)(2)Ru(II) complex (bpy=2,2'-bipyridine) have been synthesized from the tetralactam macrocycle and the rotaxanes. The anion binding abilities of the tetralactam macrocycle and its (bpy)(2)Ru(II) complex in DMSO have been compared by (1)H NMR titration experiments, which revealed significantly enhanced binding by the metal complex. Mass spectrometry has been used to study the potential formation of larger assemblies of copper(I) and the catenane built-up from two tetralactam macrocycles. Indeed, a 2:2 complex was identified. In contrast, the octalactam macrocycle of the same elemental composition yields only 1:1 complexes, with the Cu(I) ion connecting its two biquinoline moieties in the center of a figure-eight-shaped molecule. Molecular modeling studies support the structural assignments made. PMID- 12645024 TI - Structural investigation of high-valent manganese-salen complexes by UV/Vis, Raman, XANES, and EXAFS spectroscopy. AB - XANES and EXAFS spectroscopic studies at the Mn-K- and Br-K-edge of reaction products of (S,S)-(+)-N,N'-bis(3,5-di-tert-butylsalicylidene)-1,2 cyclohexanediaminomanganese(III) chloride ([(salen)Mn(III)Cl], 1) and (S,S)-(+) N,N'-bis(3,5-di-tert-butylsalicylidene)-1,2-cyclohexanediaminomanganese(III) bromide ([(salen)Mn(III)Br], 2) with 4-phenylpyridine N-oxide (4-PPNO) and 3 chloroperoxybenzoic acid (MCPBA) are reported. The reaction of the Mn(III) complexes with two equivalents of 4-PPNO leads to a hexacoordinated compound, in which the manganese atom is octahedrally coordinated by four oxygen/nitrogen atoms of the salen ligand at an average distance of approximately 1.90 A and two additional, axially bonded oxygen atoms of the 4-PPNO at 2.25 A. The oxidation state of this complex was determined as approximately +IV by a comparative study of Mn(III) and Mn(V) reference compounds. The green intermediate obtained in reactions of MCPBA and solutions of 1 or 2 in acetonitrile was investigated with XANES, EXAFS, UV/Vis, and Raman spectroscopy, and an increase of the coordination number of the manganese atoms from 4 to 5 and the complete abstraction of the halide was observed. A formal oxidation state of IV was deduced from the relative position of the pre-edge 1s-->3d feature of the X-ray absorption spectrum of the complex. The broad UV/Vis band of this complex in acetonitrile with lambda(max)=648 nm was consistent with a radical cation structure, in which a MCPBA molecule was bound to the Mn(IV) central atom. An oxomanganese(V) or a dimeric manganese(IV) species was not detected. PMID- 12645025 TI - Electronic and steric substituent influences on the conformational equilibria of cyclohexyl esters: the anomeric effect is not anomalous! AB - The cyclohexyl esters of a series of carboxylic acids, RCO(2)H, spanning a range of electronegativities and quotients of steric hindrance for the R substituent (R=Me, Et, iPr, tBu, CF(3), CH(2)Cl, CHCl(2), CCl(3), CH(2)Br, CHBr(2), and CBr(3)) were prepared. Their conformational equilibria in CD(2)Cl(2) were examined by low-temperature (1)H NMR spectroscopy to study the axial or equatorial orientation of the ester functionality with respect to the adopted chair conformation of the cyclohexane ring. The ab initio and DFT geometry optimized structures and relative free energies of the axial and equatorial conformers were also calculated at the HF/6-311G**, MP2/6-311G**, and B3LYP/6 31G** levels of theory, both in the gas phase and in solution. In the latter case, a self-consistent isodensity polarized continuum model was employed. Only by including electron correlation in the modeling calculations for the solvated molecules was it possible to obtain a reasonable correlation between DeltaG degrees (calcd) and DeltaG degrees (exp). Both the structures and the free energy differences of the axial and equatorial conformers were evaluated with respect to the factors normally influencing conformational preference, namely, 1,3-diaxial steric interactions in the axial conformer and hyperconjugation. It was assessed that hyperconjugative interactions, sigma(C-C)/sigma(C-H) and sigma*(C-O), together with a steric effect--the destabilization of the equatorial conformer with increasing bulk of the R group--were the determinant factors for the position of the conformational equilibria. Thus, because hyperconjugation is held responsible as the mitigating factor for the anomeric effect in 2-substituted, six-membered saturated heterocyclic rings, and since it is also similarly responsible, at least partly, in these monosubstituted cyclohexanes for a preferential shift towards the axial conformer, the question is therefore raised: can the anomeric effect really be construed as anomalous? PMID- 12645026 TI - Synthesis of a core trisaccharide as a versatile building block for N-glycans and glycoconjugates. AB - N-linked oligosaccharides from glycoproteins (N-glycans) can be conveniently assembled with a building block approach. A protected form of the core trisaccharide (beta-mannosyl chitobiose) was identified as a key building block. The chitobiose part of the core trisaccharide was built from a glycosyl fluoride, which served as a precursor for the reducing GlcNAc azide and the inner GlcNAc moiety. Beta-mannosylation was accomplished at the trisaccharide stage by intramolecular inversion of a beta-glucosyl chitobiose. The benzylidene protection of the beta-mannoside and the azido group at the reducing end of the core trisaccharide allow modular synthesis of N-glycans and their glycoconjugates. PMID- 12645027 TI - Time-resolved resonance Raman and density functional study of an azirine intermediate in the 2-fluorenylnitrene ring-expansion reaction to form a dehydroazepine product. AB - We report time-resolved resonance Raman spectra for the azirine intermediate produced in the 2-fluorenylnitrene ring-expansion reaction to form a dehydroazepine product. The Raman bands obtained with a 252.7 nm probe wavelength and 500 ns delay time exhibit reasonable agreement with predicted vibrational frequencies from density functional calculations for two isomers of azirine intermediates that may be formed from a 2-fluorenylnitrene precursor. The Raman bands observed for delay times of 15 ns and 10 micros were consistent with predicted vibrational frequencies from density functional calculations for the 2 fluorenylnitrene and dehydroazepine product species as well as previously reported 416 nm time-resolved Raman spectra obtained on the ns and micros time scales. Our results demonstrate that the 2-fluorenylnitrene ring-expansion reaction to produce dehydroazepine products proceeds via relatively long-lived 2 fluorenylnitrene and azirine intermediates. Substitution of a phenyl ring para to the nitrene group of phenylnitrene appears to lead to significant changes in the ring-expansion reaction so that longer lived arylnitrene and azirine intermediates can be observed. This should enable the chemical reactivity of azirine intermediates formed from arylnitrenes to be examined more readily. PMID- 12645028 TI - Identification and characterization of monomeric, volatile SiCl3NH2 as product of the reaction between SiCl4 and NH3: an important intermediate on the way to silicon nitride? AB - We studied the reaction of SiCl(4) with NH(3) by mass spectrometry and IR spectroscopy. By means of mass spectrometry, SiCl(3)NH(2) was for the first time identified as an intermediate generated in significant amounts in the course of the reaction. In additional experiments, SiCl(3)NH(2) was formed as a stable gaseous product of the ammonolysis of SiCl(4), and the product was identified and characterized in detail by IR spectroscopic methods (gas phase and matrix isolation) in combination with quantum-chemical calculations. The calculations also gave access to important thermodynamical data. PMID- 12645029 TI - Novel macrocyclic EuII complexes: fast water exchange related to an extreme M-O water distance. AB - Eu(II) complexes are potential candidates for pO(2)-responsive contrast agents in magnetic resonance imaging. In this regard, we have characterized two novel macrocyclic Eu(II) chelates, [Eu(II)(DOTA)(H(2)O)](2-) and [Eu(II)(TETA)](2-) (H(4)DOTA=1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, H(4)TETA=1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid) in terms of redox and thermodynamic complex stability, proton relaxivity, water exchange, rotation and electron spin relaxation. Additionally, solid-state structures were determined for the Sr(II) analogues. They revealed no inner-sphere water in the TETA and one inner-sphere water molecule in the DOTA complex. This hydration pattern is retained in solution, as the (17)O chemical shifts and (1)H relaxation rates proved for the corresponding Eu(II) compounds. The thermodynamic complex stability, determined from the formal redox potential and by pH potentiometry, of [Eu(II)(DOTA)(H(2)O)](2-) (lg K(Eu(II))=16.75) is the highest among all known Eu(II) complexes, whereas the redox stabilities of both [Eu(II)(DOTA)(H(2)O)](2-) and [Eu(II)(TETA)](2-) are inferior to that of 18-membered macrocyclic Eu(II) chelates. Variable-temperature (17)O NMR, NMRD and EPR studies yielded the rates of water exchange, rotation and electron spin relaxation. Water exchange on [Eu(II)(DOTA)(H(2)O)](2-) is remarkably fast (k298(ex)=2.5 x 10(9) s(-1)). The near zero activation volume (DeltaV++ =+0.1+/-1.0 cm(3) mol(-1)), determined by variable-pressure (17)O NMR spectroscopy, points to an interchange mechanism. The fast water exchange can be related to the low charge density on Eu(II), to an unexpectedly long M-O(water) distance (2.85 A) and to the consequent interchange mechanism. Electron spin relaxation is considerably slower on [Eu(II)(DOTA)(H(2)O)](2-) than on the linear [Eu(II)(DTPA)(H(2)O)](3-) (H(5)DTPA=diethylenetriaminepentaacetic acid), and this difference is responsible for its 25 percent higher proton relaxivity (r(1)=4.32 mM(-1) s(-1) for [Eu(II)(DOTA)(H(2)O)](2-) versus 3.49 mM(-1) s(-1) for [Eu(II)(DTPA)(H(2)O)](3-); 20 MHz, 298 K). PMID- 12645030 TI - A stereoselective and short total synthesis of the polyhydroxylated gamma-amino acid (-)-detoxinine, based on stereoselective preparation of dihydropyrrole derivatives from lithiated alkoxyallenes. AB - Based on our earlier results employing lithiated methoxyallene 2 as C(3) building block and imines 3 for the synthesis of dihydropyrrole derivatives 5, we have investigated chiral imines 6, 10, and 15 as electrophilic components. Combined with lithiated alkoxyallenes, these imines provide the corresponding primary adducts and finally the dihydropyrrole derivatives 8, 12, 17, 20, and 22 in good yields and with high to excellent syn selectivities. This stereochemical outcome is interpreted as a result of alpha-chelate control. Treatment with hydrochloric acid converted syn-8 and syn-12 into bicyclic compounds 9 and 13, whereas under more mildly acidic conditions adduct syn-17 was transformed into diol syn-18. The total synthesis of the uncommon gamma-amino acid (-)-detoxinine could be achieved by starting from (S)-malic acid, which was converted into imine 15 in four steps. Lithiated benzyloxyallene added to imine 15 and efficiently furnished the crucial dihydropyrrole derivative syn-22. The hydrogenolysis of this compound did not directly provide the protected triol 29 as anticipated, but a stepwise protocol made the triol available in a fairly satisfactory manner. A second crucial step of the synthesis was the selective oxidation of 29, which could be achieved by employing platinum dioxide and oxygen. The resulting bicyclic lactone 30 was smoothly transformed into enantiopure (-)-detoxinine. Thus, a fairly short synthesis of this natural product based on a lithiated alkoxyallene could be performed, demonstrating the potential of these intermediates for syntheses of interesting functionalized heterocyclic compounds. PMID- 12645031 TI - Homogeneous catalysts supported on soluble polymers: biphasic Sonogashira coupling of aryl halides and acetylenes using MeOPEG-bound phosphine-palladium catalysts for efficient catalyst recycling. AB - The Sonogashira coupling of various aryl bromides and iodides with different acetylenes was studied under biphasic conditions with soluble, polymer-modified catalysts to allow the efficient recycling of the homogeneous catalyst. For this purpose, several sterically demanding and electron-rich phosphines of the type R(P)PR(2) were synthesised. They are covalently linked to a monomethyl polyethylene glycol ether with a mass of 2000 Dalton (R(P)=MeOPEG(2000)) R(P)PR(2): -PR(2)= -CH(2)C(6)H(4)CH(2)P(1-Ad)(2), -C(6)H(4)-P(1-Ad)(2), -C(6)H(4) PPh(2). To couple aryl iodides and acetylenes, the catalyst [(MeCN)(2)PdCl(2)]/2 R(P)-C(6)H(4)-PPh(2) was used in CH(3)CN/Et(3)N/n-heptane (5/2/5). The combined yields of coupling product over five reaction cycles are between 80-95 percent. There is no apparent leaching of the catalyst into n-heptane, as evidenced by (1)H NMR spectroscopy. The new catalyst [(MeCN)(2)PdCl(2)]/2 (1-Ad)(2)PBn can be used for room-temperature coupling of various aryl bromides and acetylenes in THF with HNiPr(2) as a base. A closely related catalyst Na(2)[PdCl(4)]/2 R(P) CH(2)C(6)H(4)CH(2)P(1-Ad)(2) linked to the polymer was used to couple aryl bromides and acetylenes in DMSO or DMSO/n-heptane at 60 degrees C with 0.5 mol percent Na(2)[PdCl(4)], 1 mol percent R(P)PR(2) and 0.33 mol percent CuI. The combined yield of coupling products over five cycles is always greater than 90 percent, except for sterically hindered aryl bromides. The determination of the turnover frequency (TOF) of the catalyst indicates only a small decrease in activity over five cycles. Leaching of the catalyst into the product containing n heptane solution could not be detected by means of (1)H NMR and TXRF; this is indicative of >99.995 percent catalyst retention in the DMSO solvent. PMID- 12645032 TI - Hydrogenolysis of cycloalkanes on a tantalum hydride complex supported on silica and insight into the deactivation pathway by the combined use of 1D solid-state NMR and EXAFS spectroscopies. AB - Hydrogenolysis of cyclic alkanes is catalysed by [(triple bond)SiO)(2)Ta-H] (1) at 160 degrees C and leads to lower alkanes and cyclic alkanes including cyclopentane. The turnover number is correlated with the number of carbon atoms of the cyclic alkanes, and therefore while cycloheptane is readily transformed, cyclopentane does not give any product (<1 %). The mechanism of ring contraction probably involves carbene de-insertion as a key carbon-carbon bond-cleavage step. The reluctance of cyclopentane to undergo hydrogenolysis was further studied: under the reaction conditions cyclopentane reacts with 1 to give the corresponding cyclopentyl derivative [(triple bond)SiO)(2)Ta-C(5)H(9)] (13), which evolves towards cyclopentadienyl derivative [(triple bond)SiO)(2)Ta(C(5)H(5))] (14) according to both solid-state NMR and EXAFS spectroscopies. This latter complex is inactive in the hydrogenolysis of alkanes, and therefore the formation of cyclopentane in the hydrogenolysis of various cyclic alkanes is probably responsible for the de-activation of the catalyst by formation of cyclopentadienyl complexes. PMID- 12645033 TI - Total synthesis of mannosyl tryptophan and its derivatives. AB - Glycosylation is one of the most important post- or co-translational modifications of proteins, which affects the biological activities of the parent proteins by influencing the higher-order structure. Recently, a highly novel variant of glycoproteins that incorporate a C-glycosylated amino acid was identified in various proteins. The total synthesis of one such C-glycosyl amino acid, namely, C (2)-alpha-D-C-mannosylpyranosyl-L-tryptophan and related peptides were successfully achieved. The mannose and tryptophan moieties were connected via ring opening of benzyl-protected 1,2-anhydro-mannose by a lithiated indole derivative. After the functional group conversion and deprotection steps, the glyco-amino acid was synthesized in a concise and stereoselective manner, in high overall yields. The stereoisomer, C (2)-alpha-D-C-glycosylpyranosyl-L-tryptophan was synthesized in a similar way. Furthermore, it was revealed that the intermediate azido acid can serve as a useful building block for peptide elongation. A synthetic route for the peptide bond formation of a glycopeptide, without protection of the hydroxyl groups, using the triazine salt derivative as a coupling reagent is also reported. PMID- 12645035 TI - Progression of dopaminergic hypofunction in striatal subregions in Parkinson's disease using [18F]CFT PET. AB - The aim of this study was to investigate the progression of dopaminergic hypofunction in striatal subregions in Parkinson's disease (PD). We studied 12 patients with early PD and 11 healthy controls with a dopamine transporter ligand 2beta-carbomethoxy-3beta-(4-[(18)F]-fluorophenyl)tropane ([(18)F]CFT) positron emission tomography (PET). The PET scan was carried out twice with an average interval of 2.2 years. The regions of interest (anterior and posterior putamen, caudate nucleus, and cerebellum) were drawn on individual magnetic resonance imaging (MRI) images, matched with the PET images, and copied onto the PET images. At the first PET scan in PD patients, the [(18)F]CFT uptake in the anterior putamen was 1.92 +/- 0.67, which was 45% of the control mean, and in the posterior putamen 1.02 +/- 0.55, being only 27% of the control mean. For the caudate nucleus the corresponding figure was 2.55 +/- 0.58 (71% of the control mean). The uptake ratios had declined significantly by the time of the second PET scan and the absolute annual rate of decline of the tracer uptake was 0.23 +/- 0.14 (P < 0.001) in the anterior putamen, 0.13 +/- 0.13 (P = 0.005) in the posterior putamen, and 0.20 +/- 0.15 (P < 0.001) in the caudate nucleus. There was a statistically significant difference of the decline in the tracer uptake between the anterior and posterior putamen (P = 0.033). When the rate of progression was calculated compared to the normal control mean, the rate of annual decline was 5.3% in the anterior putamen, 3.3% in the posterior putamen, and 5.6% in the caudate nucleus, without significant changes among striatal subregions (P = 0.10). When ipsi- and contralateral sides were analyzed separately, the absolute decline of [(18)F]CFT uptake in the putamen was higher in the side ipsilateral to the predominant symptoms than in the contralateral side (P = 0.035 for anterior putamen and P = 0.026 for posterior putamen). In the caudate nucleus the absolute decline was not different between ipsi- and contralateral sides (P = 0.76). In healthy controls, no significant decline of [(18)F]CFT uptake was detected. The results are suggestive of slower progression in the posterior putamen, where the disease is more advanced, but studies to follow up the same patient at several time points are needed to resolve this question. Synapse 48:109-115, 2003. PMID- 12645036 TI - Influence of acetylcholine levels on the binding of a SPECT nicotinic acetylcholine receptor ligand [123I]5-I-A-85380. AB - Although in vitro theory indicates that ligand binding is sensitive to competition with neurotransmitters, only some imaging ligands have shown such competition in vivo. The purpose of this study was to determine whether increases in acetylcholine (ACh) levels induced by an acetylcholinesterase inhibitor, physostigmine, inhibit in vivo binding of [(123)I]5-iodo-3-(2(S)-2-azetidinyl methoxy) pyridine (5-I-A-85380), a single photon emission computed tomography ligand for the high-affinity type nicotinic ACh receptor (nAChR). Baboons were used for seven studies with a bolus plus constant infusion equilibrium paradigm. After achieving equilibrium at 5 h, physostigmine (0.02 (n = 1), 0.067 (n = 3), and 0.2 (n = 3) mg/kg) was administered intravenously and data were acquired for up to 8 h. To confirm equilibrium conditions, [(123)I]5-I-A-85380 plasma levels were measured in four studies, including all studies with 0.2 mg/kg physostigmine. Prior to physostigmine administration, thalamic activities were stable, with changes of 1.1%/h or less, except in one study with a gradual increase of 4.2%/h. Thalamic activities were decreased by 15% in one study with 0.067 mg/kg and 14-17% in all studies with 0.2 mg/kg physostigmine administration (P = 0.009). In these studies with 0.2 mg/kg physostigmine administration, [(123)I]5-I-A-85380 plasma levels showed a transient or a sustained increase after physostigmine administration that would have increased thalamic activities. These results suggest that elevated ACh levels induced by physostigmine can effectively compete in vivo with [(123)I]5-I-A-85380 binding at nAChRs. However, decreased thalamic activities could have been caused by other mechanisms, including internalization of the receptor with an associated decreased affinity for radioligand. PMID- 12645037 TI - Actions of the sodium channel inhibitor 202W92 on rat midbrain dopaminergic neurons. AB - Excessive glutamatergic activity is implicated in Parkinson's disease (PD) and sodium channel blockade, resulting in inhibition of glutamate release, is a potential therapeutic approach to PD therapy. Beneficial effects of riluzole and lamotrigine have been reported in animal models of PD, but these compounds have relatively low potency as sodium channel inhibitors and also inhibit N and P/Q type calcium channels. 202W92, a structural analog of lamotrigine, is a potent sodium channel inhibitor, with no effect on N, P/Q-type channels. Here we present the effects of 202W92 on single patch-clamped dopaminergic neurons. 202W92 (> or =10 microM) inhibited spontaneous action potential firing and reduced amplitude and frequency of evoked action potentials. It also inhibited the frequency of 4 aminopyridine (4-AP)- and electrically evoked excitatory postsynaptic currents (EPSCs) and GABAergic inhibitory postsynaptic currents (IPSCs), with >80% inhibition at 10 microM (IC(50) 1.5 microM). EPSC and IPSC amplitudes were partially inhibited. 202W92 did not affect postsynaptic responses to locally applied glutamate and GABA, nor spontaneously occurring mini-IPSCs. These actions of 202W92 are compatible with sodium channel inhibition and depression of transmitter release. PMID- 12645038 TI - Methamphetamine exposure during the preweanling period causes prolonged changes in dorsal striatal protein kinase A activity, dopamine D2-like binding sites, and dopamine content. AB - Exposure to methamphetamine (METH) during the preweanling period produces few, if any, neurotoxic effects (using criteria established in adult rats), yet it has substantial long-term effects on a variety of behavioral measures (e.g., locomotor activity, acoustic startle response, and spatial learning). The purpose of the present study was to examine the long-term changes in dopaminergic functioning brought about by early METH exposure. Rats were injected with METH (10 mg/kg) or saline four times daily on postnatal days (PD) 11-20 and housed undisturbed until PD 90, at which time they were killed and their dorsal striata (i.e., caudate-putamen) were removed and frozen for assay. The ability of early METH exposure to alter protein kinase A (PKA) activity and dopamine (DA) D(2) like binding sites, as well as DA and DOPAC content, were assessed. Results showed that METH exposure on PD 11-20 caused long-term reductions in all of the dopaminergic markers assayed. METH-induced reductions in DA content and D(2)-like receptors were observed. Some sex differences were apparent, as the METH-induced decreases in PKA activity and DOPAC content were more evident in male rats. In conclusion, preweanling METH exposure caused changes in DA markers that were still detectable at PD 90; however the magnitude of many of these effects (e.g., the reductions in DA and DOPAC levels) was substantially less than typically reported for rats treated with METH in adulthood. The ability of METH to cause long-term reductions in PKA activity may partially account for some of behavioral deficits exhibited by rats exposed to METH prior to weaning. PMID- 12645039 TI - Developmental D-methamphetamine treatment selectively induces spatial navigation impairments in reference memory in the Morris water maze while sparing working memory. AB - In previous studies, we have shown that P11-20 treatment with D-methamphetamine (MA) (10 mg/kg x 4/day at 2-h intervals) induces impairments in spatial learning and memory in the Morris water maze after the offspring reach adulthood. Using a split-litter, multiple dose, design (0, 5, 10, and 15 mg/kg MA administered s.c. 4/day at 2-h intervals), the spatial learning effect was further explored with a multiple shifted platform (reversal), reference memory-based procedure and a working memory procedure. Prior to spatial learning, animals were first tested for swimming ability (in a straight swimming channel), sequential learning (in the Cincinnati multiple-T water maze), and proximal cue learning (in the Morris water maze). Rats were then assessed in the hidden platform, reference memory based spatial version of the Morris maze for acquisition and on five subsequent phases in which the platform was moved to new locations. After the reference memory-based, fixed platform position learning phases, animals were tested in the trial-dependent, matching-to-sample, working memory version of the Morris maze. No group differences were found in straight channel, sequential maze, or cued Morris maze performance. By contrast, all MA groups were impaired in spatial learning during acquisition, multiple shift, and shifted with a reduced platform phases of reference memory-based learning. In addition, MA animals were impaired on memory (probe) trials during the acquisition and shifted with a reduced platform phases of learning. No effects on trial-dependent, matching-to-sample, working memory were found. The findings demonstrate that neonatal treatment with MA induces a selective impairment of reference memory-based spatial learning while sparing sequential, cued, and working memory-based learning. PMID- 12645040 TI - Environmental complexity has different effects on the structure of neurons in the prefrontal cortex versus the parietal cortex or nucleus accumbens. AB - Complex housing has been used widely as a model of experience-dependent change. Animals housed in complex environments typically show synaptogenesis throughout the sensory and motor cortex as well as the striatum and hippocampus, and thus it is generally assumed that such changes are likely to be found throughout the cerebrum. The purpose of the present study was to determine whether persistent alterations of dendritic morphology would be found in two regions that had previously not been examined, namely, the medial prefrontal region (Cg3) and nucleus accumbens (NAcc). The results show that housing female rats in complex environments for 3.5 months increased dendritic arborization on medium spiny neurons in the NAcc and on pyramidal cells in the somatosensory cortex (Par 1), but not in Cg3. Environmental complexity increased spine density in all three areas, however. The failure to find increased dendritic length or branching in Cg3 was unexpected. Thus, the data suggest that complex housing may not engage prefrontal neurons in the same manner as neurons in sensory or motor areas. It appears that complex housing may not produce generalized changes in cerebral morphology. The data further suggest that it is prudent to measure both dendritic length and spine density in studies of experience-dependent effects on synaptic plasticity. PMID- 12645041 TI - Acute administration of the selective D3 receptor antagonist SB-277011A blocks the acquisition and expression of the conditioned place preference response to heroin in male rats. PMID- 12645042 TI - The competency of criminal defendants to refuse, for delusional reasons, a viable insanity defense recommended by counsel. AB - This article addresses the issues of whether mentally ill defendants charged with serious crimes who refuse to plead a viable and counsel-recommended insanity defense for delusional reasons (but who are otherwise competent to stand trial) should be considered to be competent, or incompetent, to stand trial; whether such defendants should be allowed to represent themselves with a delusional defense; and whether an insanity defense may properly be imposed upon such defendants. Based on an analysis of relevant Supreme Court decisions and other relevant cases, it is concluded that such defendants should not be allowed to go forward with a delusional defense (at least until reasonable efforts to treat the defendants' delusions are made). It is also argued, however, that unless an insanity defense would be viable (as well as recommended by counsel) delusional defendants who are otherwise competent to stand trial should be permitted to go forward, and represent themselves, with the defense of their choosing. PMID- 12645043 TI - Differentiating sex offences: a behaviorally based thematic classification of stranger rapes. AB - It is hypothesized that stranger rape victim statements will reveal a scale of violation experienced by the victim, ranging from personal violation, through to physical violation, and finally, at the most extreme level, sexual violation. It is also hypothesized that offences can be differentiated in terms of one of four themes: hostile, controlling, stealing, or involving. To test these hypotheses, crime scene data from 112 rapes were analyzed by the multi-dimensional scaling procedure Smallest Space Analysis. The results provide empirical support for a composite model of rape consisting of four behavioral themes as different expressions of various intensities of violation. The results also suggest that stranger rapes may be less about power and control than about hostility and pseudo-intimacy. The proposed model has implications for the classification of rape, the investigation of sexual crimes, and the treatment of victims. PMID- 12645044 TI - Effectiveness of participation as a defendant: the attorney-juvenile client relationship. AB - Recent changes in the processing of juveniles in the justice system place greater significance on children's capacities to participate in legal contexts. Effective participation as a defendant encompasses abilities beyond those legally required for adjudicative competence, which may nevertheless influence the quality and nature of a defendant's participation in the trial process. Based in developmental judgment theory, the current study compares 203 juveniles and 110 adults detained pre-trial using a hypothetical attorney-client vignette to examine how psychosocial factors are reflected in decision-making processes and link to decision outcomes and effective participation within the attorney-client relationship. Age-related differences in legally relevant decision-making processes and outcomes are identified, and implications for policy are made. PMID- 12645046 TI - Juror decision-making in a mock sexually violent predator trial: gender differences in the impact of divergent types of expert testimony. AB - Despite widespread use of mental health testimony in cases where violence risk is at issue, relatively little is known about the impact of such information on juror decision-making. This study addressed the effects of testimony based on three types of risk assessment instrument or method (clinical opinion, actuarial assessment, and ratings of psychopathy) to examine whether they would have differential impact on jurors' perceptions of the defendant. In a mock sexually violent predator civil commitment trial, 172 undergraduates were presented a case summary that included prosecution and defense expert testimony related to violence risk based on one of the three methods noted above. Consistent with earlier research, the hypothesis that a defendant described as a "high risk psychopath" by the prosecution would be judged more severely than a defendant judged as "high risk" based on other evaluation procedures was supported, but only among female jurors. Unlike prior studies, little support was found for the hypothesis that clinical opinion testimony would be more influential than actuarially based testimony for either gender. Mechanisms that may underlie the observed gender differences are discussed, as are the potential implications of these findings for civil commitment proceedings. PMID- 12645045 TI - Gender and role-based perceptions of domestic abuse: does sexual orientation matter? AB - Although it is estimated that domestic abuse is as common in gay male and lesbian intimate relationships as in heterosexual relationships, the legal system often fails to recognize or respond to same-gender cases. Empirical research examining the impact of sexual orientation on perceptions of abuse is virtually nonexistent. Undergraduates (N = 252) read a summary of a domestic abuse incident in which victims and perpetrators varied by gender and, by implication, sexual orientation. Victim and respondent gender, rather than the couple's sexual orientation, primarily affected responses to domestic abuse. Domestic abuse perpetrated against women was perceived to be more serious and in need of intervention than abuse against men. Women were more likely than men to believe the victim and to recommend criminal justice system interventions. Because they are inconsistent with gender role stereotypes, domestic abuse cases involving male victims or female perpetrators may not receive equitable treatment within the criminal justice system. PMID- 12645047 TI - The relationship of mental illness to targeted contact behavior toward state government agencies and officials. AB - Research in the burgeoning field of threat assessment has illuminated the importance of mental illness factors when considering risk of targeted violence particularly related to government agencies and officials. The authors analyzed 127 cases investigated by a state law enforcement agency regarding threatening or other contacts toward public officials or state agency employees prompting security intervention. Univariate and discriminant analysis indicated that mentally ill subjects were significantly more likely to engage in more contacts as well as to make specific demands during such contacts. Mentally ill subjects were also more likely to articulate help-seeking concerns and employ religious themes, as opposed to using insulting, degrading, or ominous language toward the target or issuing complaints regarding policy issues. Contrary to other research, the mentally ill subjects within this sample were not significantly more likely to engage in approach behavior, a threshold for higher risk of violence. Implications for threat assessment activity are discussed. PMID- 12645052 TI - Supercarboranes. PMID- 12645048 TI - Narrative plausibility: the impact of sequence and anchoring. AB - The perceived plausibility of suspect narratives is hypothesized to be a product of more than logical evaluation. Aspects of the narrative's internal structure, notably the extent to which it follows a canonical (or stereotypical) sequence of events, may influence judged plausibility. Plausibility may also be sensitive to external "anchors" that activate relevant schema. To test these possibilities, variations of two suspect testimonies were created in accordance with the model by Stein and Glenn (1979) of a stereotypical story grammar, and the account by Wagenaar, van Koppen, and Crombag (1993) of narrative anchoring. Subjects rated the narrative account using a perceived plausibility scale developed from pilot work. ANOVA revealed that criminal anchoring in suspect statements, regardless of the crime scenario, has a negative effect on the plausibility level. Similarly, plausibility levels were lower when the statement did not follow a temporal sequence of events. The implications for models of how people judge plausibility are discussed, as are the practical implications for legal contexts. PMID- 12645049 TI - Sex offenders and actuarial risk assessments: ethical considerations. AB - When assessing the recidivism risk of previously convicted sex offenders, the relevant ethical standards and practice guidelines obligate psychologists to acknowledge numerous limits related to their data and conclusions. For the actuarial instruments used in these assessments, the highest rates of classification accuracy are associated with greater specificity compared with sensitivity. Selecting cut-off scores to maximize sensitivity results in an inordinate frequency of false positive classifications. Attempts at maximizing specificity create an undesirable frequency of false negative classifications.Unfortunately, adjusted actuarial assessment cannot remedy these problems created by the sensitivity-specificity tradeoff. As an unstandardized procedure, the accuracy of adjusted actuarial assessment is severely limited. Consequently, civil proceedings for previously convicted sexual offenders rely on assessments of very limited accuracy. Undertaking these assessments, therefore, may be inconsistent with relevant ethical standards and practice guidelines. PMID- 12645053 TI - The stories of santonin and santonic acid. PMID- 12645055 TI - 2,2'-diborabiphenyl: a Lewis acid analogue of 2,2'-bipyridine. PMID- 12645054 TI - Interactions with aromatic rings in chemical and biological recognition. AB - Intermolecular interactions involving aromatic rings are key processes in both chemical and biological recognition. Their understanding is essential for rational drug design and lead optimization in medicinal chemistry. Different approaches-biological studies, molecular recognition studies with artificial receptors, crystallographic database mining, gas-phase studies, and theoretical calculations-are pursued to generate a profound understanding of the structural and energetic parameters of individual recognition modes involving aromatic rings. This review attempts to combine and summarize the knowledge gained from these investigations. The review focuses mainly on examples with biological relevance since one of its aims it to enhance the knowledge of molecular recognition forces that is essential for drug development. PMID- 12645056 TI - A convergent total synthesis of phorboxazole A. PMID- 12645058 TI - Simple, three-dimensional microfabrication of electrodeposited structures. PMID- 12645059 TI - An unconventional cobalt-catalyzed aerobic oxidation of tertiary nitrogen compounds to N-oxides. PMID- 12645057 TI - Total synthesis of phorboxazole A. PMID- 12645060 TI - Unified strategy for the synthesis of (-)-elisapterosin B and (-)-colombiasin A. PMID- 12645061 TI - Selection and amplification of a catalyst from a dynamic combinatorial library. PMID- 12645062 TI - Highly enantioselective Alk-2-enylation of aldehydes through an allyl-transfer reaction. PMID- 12645063 TI - Cu-catalyzed enantioselective conjugate additions of alkyl zinc reagents to unsaturated N-acyloxazolidinones promoted by a chiral triamide phosphane. PMID- 12645065 TI - SPANphos: a C2-symmetric trans-coordinating diphosphane ligand. PMID- 12645066 TI - Rate enhancement with a bowl-shaped phosphane in the rhodium-catalyzed hydrosilylation of ketones. PMID- 12645064 TI - Total synthesis as a resource in the discovery of potentially valuable antitumor agents: cycloproparadicicol. PMID- 12645067 TI - Norbadione A: synthetic approach to the bis(pulvinic acid) moiety and cesium complexation studies. PMID- 12645068 TI - Large parity-violation effects in heavy-metal-containing chiral compounds. PMID- 12645069 TI - Structural, electronic, and impurity-doping effects in nanoscale chemistry: supported gold nanoclusters. PMID- 12645070 TI - Optimized NMR spectroscopic method for the configurational analysis of chemically equivalent vicinal protons. PMID- 12645071 TI - A novel class of aromatic compounds: metal-centered planar cations [Fe(Sb5)]+ and [Fe(Bi5)]+. PMID- 12645072 TI - Highly reactive polyisobutenes prepared with manganese(II) complexes as initiators. PMID- 12645074 TI - Relationship between cell cycle molecules and onset of oligodendrocyte differentiation. PMID- 12645075 TI - Genetic background determines phenotypic severity of the Plp rumpshaker mutation. AB - The rumpshaker mutation of the proteolipid protein (Plp) gene causes dysmyelination in man and mouse. We show that the phenotype in the mouse depends critically on the genetic background in which the mutation is expressed. On the C3H background there is normal longevity whereas changing to a C57BL/6 strain results in seizures and death at around postnatal day 30. The more severe phenotype is associated with less myelin and reduced levels of major myelin proteins. There are also more apoptotic cells, including oligodendrocytes, increased numbers of proliferating cells, increased numbers of NG2+ oligodendrocyte progenitors and increased microglia compared to the milder phenotype. The number of mature oligodendrocytes is similar to wild-type in both strains of mutant, however, suggesting that increased oligodendrocyte death is matched by increased generation from progenitors. The dichotomy of phenotype probably reflects the influence of modifying loci. The localization of these putative modifying genes and their mode of action remain to be determined. PMID- 12645078 TI - Involvement of the alpha 7 subunit of the nicotinic receptor in morphogenic and trophic effects of acetylcholine on embryonic rat spinal motoneurons in culture. AB - The morphogenic and trophic effects of acetylcholine (ACh) on embryonic cultured rat spinal cord motoneurons (MNs) through nicotinic alpha7 autoreceptors were assessed. Alpha7 Subunits of the nicotinic cholinergic receptor were detected in cultures of purified rat spinal embryonic MNs sampled at E15, by both immunocytochemistry and alpha-bungarotoxin binding. According to these two methods, alpha7 subunits are located mainly at somatic and axonal membrane. Functional involvement of the alpha7 subunit in survival and development of morphological properties of growing cultured MNs was tested using an antisense strategy. The antisense oligonucleotide significantly decreases the expression of the alpha7 protein compared with control and mismatch oligonucleotide-treated cultures. This decrease in the expression of the alpha7 protein leads to a significant increase in the number of axonal branches and in the length of the axon. The antisense treatment also induces, as early as the first day in culture, a decrease of MN survival, leading to total cell death at day 5. TUNEL staining revealed that the MNs are dying through apoptotic processes. Thus, our study shows that ACh is a morphogenic and trophic factor. These effects are directly linked to the membrane expression level of alpha7 protein. Indeed, the lower the alpha7 expression, the lower the inhibition of axonal growth (i.e., axonal elongation) and the lower the MN survival. PMID- 12645076 TI - Effect of hypoglycemia on brain glycogen metabolism in vivo. AB - The brain contains a small but significant amount of glycogen, which has long been considered to play an insignificant role in the brain. In this study, brain glycogen metabolism was measured using (13)C NMR spectroscopy at 9.4 T. Brain glycogen metabolism was modulated by hyperinsulinemia resulting in a net accumulation. The role of glycogen in maintaining brain function is unknown; one possibility is that it may serve as an endogenous glucose reservoir to protect the brain against severe hypoglycemia. To address this possibility, rats were subjected to insulin-induced moderate hypoglycemia and when the level of brain glucose approached zero, brain glycogen content began to decrease gradually, demonstrating utilization of this glucose reservoir. The brain glycogen signal never became undetectable, however, even during 2 hr of hypoglycemia. When plasma and brain glucose concentrations were restored, glycogen increased and the concentration exceeded the pre-hypoglycemic level by several-fold. The data suggest that brain glycogen can provide fuel for extended periods of time when glucose supply is inadequate. Furthermore, brain glycogen can rebound (super compensate) after a single episode of hypoglycemia. We postulate that brain glycogen serves as an energy store during hypoglycemia and that it may participate in the creation of reduced physiological responses to hypoglycemia that are involved in a symptom often observed in patients with diabetes, hypoglycemia unawareness. PMID- 12645077 TI - Regulation of FLRG expression in rat primary astroglial cells and injured brain tissue by transforming growth factor-beta 1 (TGF-beta 1). AB - Follistatin-related gene (FLRG) is a member of the follistatin family of proteins and interacts with transforming growth factor (TGF) superfamily proteins like follistatin. To understand the expression level of FLRG in brain tissue, we examined whether primary neurons and glial cells from rat embryos express FLRG mRNA and produce its protein product. FLRG and follistain mRNAs were mainly expressed in astroglial cells, while activin A mRNA was abundant in primary neurons. TGF-beta1 highly enhanced expression levels of FLRG mRNA in astroglial cells, compared with those of follistatin and activin A mRNAs. Particularly, TGF beta1 facilitated the secretion of FLRG protein from primary astroglial cells in a dose-dependent manner. Moreover, changes in expression levels of FLRG mRNA and protein in brain tissue were also analyzed after a penetrating injury, using quantitative polymerase chain reactin (PCR) and immunohistochemical methods. Expression levels of FLRG mRNA were significantly increased in damaged regions after penetrating injury together with those of activin A and TGF-beta1 mRNAs. Immunohistochemical observations showed that positive signals of FLRG protein were colocalized in glial fibrillary acidic protein-positive reactive astroglial cells located in damaged regions after a penetrating injury. The expression of follistatin mRNA rather decreased in damage regions after the brain injury. These results suggest that FLRG is synthesized in and secreted from astroglial cells. In particular, FLRG, but not follistatin, may play a role in the regulation of activin A in brain wound healing in response to TGF-beta1. PMID- 12645079 TI - Adenoviral gene transfer of GDNF, BDNF and TGF beta 2, but not CNTF, cardiotrophin-1 or IGF1, protects injured adult motoneurons after facial nerve avulsion. AB - We examined neuroprotective effects of recombinant adenoviral vectors encoding glial cell line-derived neurotrophic factor (GDNF), brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), cardiotrophin-1 (CT1), insulin like growth factor-1 (IGF1), and transforming growth factor-beta2 (TGFbeta2) on lesioned adult rat facial motoneurons. The right facial nerves of adult Fischer 344 male rats were avulsed and removed from the stylomastoid foramen, and adenoviral vectors were injected into the facial canal. Animals avulsed and treated with adenovirus encoding GDNF, BDNF, CNTF, CT1, IGF1 and TGFbeta2 showed intense immunolabeling for these factors in lesioned facial motoneurons, respectively, indicating adenoviral induction of the neurotrophic factors in these neurons. The treatment with adenovirus encoding GDNF, BDNF, or TGFbeta2 after avulsion significantly prevented the loss of lesioned facial motoneurons, improved choline acetyltransferase immunoreactivity and prevented the induction of nitric oxide synthase activity in these neurons. The treatment with adenovirus encoding CNTF, CT1 or IGF1, however, failed to protect these neurons after avulsion. These results indicate that the gene transfer of GDNF and BDNF and TGFbeta2 but not CNTF, CT1 or IGF1 may prevent the degeneration of motoneurons in adult humans with motoneuron injury and motor neuron diseases. PMID- 12645080 TI - Ceramide in rafts (detergent-insoluble fraction) mediates cell death in neurotumor cell lines. AB - Detergent-resistant lipid microdomains (Rafts) were isolated from human oligodendroglioma (HOG), human neuroblastoma (LA-N-5), and immortalized dorsal root ganglion (F-11) cell lines by sucrose-density gradient ultracentrifugation and shown to be enriched in cholesterol, sphingomyelin, and ceramide. [(3)H]palmitate labeling allowed the Raft fraction to be easily identified as a sharp peak of (3)H radioactivity in the 5-30% sucrose interphase. Treatment of [(3)H]palmitate-labeled cells with staurosporine (to activate caspase 8 and induce apoptosis) or exogenous sphingomyelinase specifically increased the [(3)H]ceramide content of the Raft fraction. Depletion of cholesterol with beta methylcyclodextran decreased Raft formation and partially blocked staurosporine induced apoptosis. Similarly, treatment of cells with Fumonisin B1 to inhibit de novo sphingolipid synthesis by 50% reduced the labeling of the Raft fraction and partially blocked staurosporine-induced apoptosis. Staurosporine treatment activated neutral sphingomyelinase but had no effect on acid sphingomyelinase activity or on other lysosomal hydrolases, such as alpha-L-fucosidase. Most of the neutral sphingomyelinase activity is in the Raft fraction, suggesting that the conversion of sphingomyelin to ceramide in Rafts is an important event in neural cell apoptosis. PMID- 12645081 TI - Interactions among nitric oxide and Bcl-family proteins after MPP+ exposure of SH SY5Y neural cells I: MPP+ increases mitochondrial NO and Bax protein. AB - We studied effects of methylpyridinium ion (MPP(+)) on apoptosis, cell death and regulation of Bcl-2-family proteins in SH-SY5Y neuroblastoma cells. MPP(+) increased intracellular accumulation of DNA-histone complexes as a measure of apoptosis and decreased intracellular calcein fluorescence as a measure of cell death. If ATP synthesis was supported, MPP(+) caused apoptosis in rho(0) cells devoid of electron transport function. Caspase inhibition blocked apoptosis but not cell death caused by MPP(+). MPP(+) increased levels of Bax, Bcl-2 and Bcl X(L) proteins approximately 2-fold over 24 hr, with Bax increases occurring first; Bax did not increase in rho(0) cells. The Bax increase, but not that of Bcl-2 or Bcl-X(L), was dependent on nitric oxide (NO) and seemed post transcriptional. DAF-FM imaging revealed increased mitochondrial NO within hours of exposure to MPP(+). Western blots showed a constitutive approximately 130 kD protein that stained for NOS-2, consistent with reports of mitochondrial nitric oxide synthase (mtNOS). MPP(+) caused a NO-dependent release of cytochrome C into cytoplasm. MPP(+) increases mitochondrial NO levels and causes a NO-dependent increase in Bax protein, providing a mechanism for NOS-and Bax-dependency of MPTP neurotoxicity in vivo and implicating locally produced NO as a signaling molecule used by mitochondria to manipulate cell death cascades. PMID- 12645082 TI - Interactions among nitric oxide and Bcl-family proteins after MPP+ exposure of SH SY5Y neural cells II: exogenous NO replicates MPP+ actions. AB - In the preceding companion article, we showed that the neurotoxin methylpyridinium (MPP(+)) increases mitochondrial nitric oxide (NO), causes a post-transcriptional, NO-dependent increase in Bax protein and produces caspase dependent apoptosis and caspase-independent cell death. In the present study, we show that exogenous NO replicates these findings. The long-term NO generator diethylenetriamine-NO (DETA-NO) reproduced the post-transcriptional Bax protein increase, but did not increase Bcl-2 or Bcl-X(L) proteins. Like MPP(+), DETA-NO caused an early decrease in Bcl-2 mRNA, did not increase Bax protein in rho(0) cells and caused caspase- and cycloheximide-dependent apoptosis and caspase independent cell death. We developed cell lines with inducible overexpression of Bcl proteins, at levels relevant to those we found in cells exposed to MPP(+) or DETA-NO. Inducible overexpression ( approximately 2-fold) of Bcl-2 or Bcl-X(L) proteins reduced MPP(+) or NO-induced apoptosis but did not affect cell death. Inducible Bax overexpression ( approximately 5-fold) slightly increased cell death. Our results show that exogenous NO mimics actions of MPP(+) on SH-SY5Y neuroblastoma cells and supports the mediation of MPP(+) neurotoxicity by NO generated intracellularly in mitochondria. PMID- 12645083 TI - Upregulation of catecholamine biosynthetic enzymes by nitric oxide. AB - Nitric oxide (NO) is recognized as an essential intercellular messenger in central and peripheral nervous systems. In the present study, whether NO exerts effects on catecholamine (CA) biosynthetic enzymes was determined in primary cultured bovine chromaffin cells. The NO generators sodium nitroprusside (SNP) and S-nitroso-N-acetyl-D,L-penicillamine, in a dose-dependent manner, upregulated transcript levels of tyrosine hydroxylase, dopamine beta-hydroxylase, and phenylethanolamine N-methyltransferase, accompanied by long-term increases in their enzyme activities and the intracellular CA levels. The SNP effect was diminished by co-treatment with LY83583, an inhibitor of soluble guanylate cyclase, or H-8, a cyclic GMP (cGMP)-dependent protein kinase inhibitor. Co treatment with 8-Br-cGMP did not increase further the expression of these enzyme genes induced by SNP. Taken together, the data suggest that NO leads to long-term upregulation of the CA system via induction of the genes involved and that this is mediated by cGMP-dependent signaling pathway. PMID- 12645086 TI - Increased nuclear factor-kappa B p65 immunoreactivity following retinal ischemia and reperfusion injury in mice. AB - Nuclear factor-kappaB (NF-kappaB) is a universal transcription factor and has previously been demonstrated to play an important role in CNS injury. This study investigated the expression of NF-kappaB in the inner layers of the retina in mice after retinal ischemia and reperfusion injury. Retinal ischemia was induced by elevation of intraocular pressure to 120 mmHg for 60 min. To evaluate inner retinal degeneration, the inner retinal thickness was quantified with an image analysis system. The inner retinal thickness increased in the initial 24 hr after retinal ischemia and was ascribed to tissue edema but was significantly decreased in the ensuing 7 days. Immunohistochemistry using NF-kappaB p65 monoclonal antibody was performed on the retina and was corelated with TUNEL labeling. Six hours after retinal ischemia, nuclear p65 immunoreactivity was increased in the inner nuclear and ganglion cell layers and reached a peak at 24 hr. The increased NF-kappaB p65 immunolabeling was parallel to the TUNEL labeling. Double labeling with p65 and TUNEL showed partial colocalization of p65 and TUNEL labeling in the scattered cells of the inner nuclear and ganglion cell layers. However, several p65-positive cells were TUNEL negative, suggesting that these cells might have survived the injury. The NF-kappaB p65 immunoreactivity was associated with retinal degeneration following retinal ischemia and reperfusion injury. PMID- 12645085 TI - L-phenylalanine selectively depresses currents at glutamatergic excitatory synapses. AB - To explore the hypothesis that L-phenylalanine (L-Phe) depresses glutamatergic synaptic transmission and thus contributes to brain dysfunction in phenylketonuria (PKU), the effects of L-Phe on spontaneous and miniature excitatory postsynaptic currents (s/mEPSCs) in rat and mouse hippocampal and cerebrocortical cultured neurons were studied using the patch-clamp technique. L Phe depressed the amplitude and frequency of both N-methyl-D-aspartate (NMDA) and non-NMDA components of glutamate receptor (GluR) s/mEPSCs. The IC(50) of L-Phe to inhibit non-NMDAR mEPSC frequency was 0.98 +/- 0.13 mM, a brain concentration seen in classical PKU. In contrast, D-Phe had a significantly smaller effect, whereas L-leucine, an amino acid that competes with L-Phe for brain transporter, had no effect on mEPSCs. Unlike GluR s/mEPSCs, GABA receptor mIPSCs were not attenuated by L-Phe. A high extracellular concentration of glycine prevented the attenuation by L-Phe of NMDAR current, activated by exogenous agonist, and of NMDAR s/mEPSC amplitude, but not of NMDAR s/mEPSC frequency. On the other hand, L Phe significantly depressed non-NMDAR current activated by low but not high concentrations of exogenous agonists. Glycine-independent attenuation of NMDAR s/mEPSC frequency suggests decreased presynaptic glutamate release caused by L Phe, whereas decreased amplitudes of NMDAR and non-NMDAR s/mEPSCs are consistent with competition of L-Phe for the glycine- and glutamate-binding sites of NMDARs and non-NMDARs, respectively. The finding that GluR activity is significantly depressed at conditions characteristic of classical PKU indicates a potentially important contribution of impaired GluR function to PKU-related mental retardation and provides important insights into the potential physiological consequences of impaired GluR function. PMID- 12645084 TI - Tyrosine hydroxylase mRNA and protein are down-regulated by chronic clozapine in both the mesocorticolimbic and the nigrostriatal systems. AB - The dopaminergic system is one of the most important targets for pharmacological treatment of schizophrenia. Despite substantial work on mechanisms of action, it is not clear which dopaminergic pathways mediate the therapeutic effects of antipsychotic drugs. It has been shown that chronic clozapine, an atypical antipsychotic, decreases dopamine levels in the mesocorticolimbic system but not in the nigrostriatal system. Because tyrosine hydroxylase is the rate-limiting enzyme in the biosynthesis of dopamine, we studied the effect of chronic clozapine in both dopaminergic systems. We demonstrated a decrease of tyrosine hydroxylase mRNA not only in the ventral tegmental area but also in the substantia nigra, the cell body areas of the mesocorticolimbic and the nigrostriatal systems, respectively. The reduced tyrosine hydroxylase mRNA level in these areas is accompanied by an ample reduction in the tyrosine hydroxylase protein level in their corresponding axonal terminal fields, the nucleus accumbens and the striatum. There was thus discordance between the clozapine induced decrease of tyrosine hydroxylase mRNA and protein and the absence of an effect on dopamine levels in the nigrostriatal system. It has been suggested that reduced levels of dopamine in the mesocorticolimbic system are required for the antipsychotic effect of the drug. Therefore, the modulation of tyrosine hydroxylase gene expression by clozapine in the mesocorticolimbic system might be necessary for its antipsychotic effect; this effect might be of relevance when considering new atypical agents. PMID- 12645087 TI - Altered levels of cerebrospinal fluid reelin in frontotemporal dementia and Alzheimer's disease. AB - Reelin is an essential glycoprotein for correct cytoarchitectonic organization during CNS development. Its function in the adult brain is far less well understood, but altered brain and blood reelin levels have been reported in some psychiatric disorders, and the possibility has been considered of an involvement of the reelin signaling pathway in neurodegeneration. Here we report, for the first time, the presence of detectable levels of reelin in rat and human cerebrospinal fluid (CSF) and show evidence for the involvement of a 180-kDa reelin fragment in two neurodegenerative disorders. This fragment was analyzed by Western blotting in CSF samples from 13 healthy control individuals and 14 frontotemporal dementia (FTD) and 20 Alzheimer's disease (AD) patients. Increased CSF 180-kDa reelin was found in FTD (161.7 +/- 6.7 arbitrary units; a.u.) and AD (151.4 +/- 3.8 a.u.) compared with control individuals (141.4 +/- 1.2 a.u., P < 0.05). Our results strongly suggest the involvement of reelin signaling in neurodegenerative pathologies. PMID- 12645089 TI - Nanoscopic aspects of radiobiological damage: Fragmentation induced by secondary low-energy electrons. AB - Low-energy electrons (LEEs) are produced in large quantities in any type of material irradiated by high-energy particles. In biological media, these electrons can fragment molecules and lead to the formation of highly reactive radicals and ions. The results of recent experiments performed on biomolecular films bombarded with LEEs under ultra-high vacuum conditions are reviewed in the present article. The major type of experiments, which measure fragments produced in such films as a function of incident electron energy (0.1-45 eV), are briefly described. Examples of the results obtained from DNA films are summarized along with those obtained from the fragmentation of elementary components of the DNA molecule (i.e., thin solid films of H(2)O, DNA bases, sugar analogs, and oligonucleotides) and proteins. By comparing the results of these different experiments, it is possible to determine fundamental mechanisms that are involved in the dissociation of biomolecules and the production of single- and double strand breaks in DNA, and to show that base damage is dependent on the nature of the bases and on their sequence context. Below 15 eV, electron resonances (i.e., the formation of transient anions) play a dominant role in the fragmentation of all biomolecules investigated. These transient anions fragment molecules by decaying into dissociative electronically excited states or by dissociating into a stable anion and a neutral radical. These fragments usually initiate other reactions with nearby molecules, causing further chemical damage. The damage caused by transient anions is dependent on the molecular environment. PMID- 12645088 TI - Contributions of mass spectrometry in the study of nucleic acid-binding proteins and of nucleic acid-protein interactions. AB - Nucleic-acid-protein (NA-P) interactions play essential roles in a variety of biological processes-gene expression regulation, DNA repair, chromatin structure regulation, transcription regulation, RNA processing, and translation-to cite only a few. Such biological processes involve a broad spectrum of NA-P interactions as well as protein-protein (P-P) interactions. These interactions are dynamic, in terms of the chemical composition of the complexes involved and in terms of their mere existence, which may be restricted to a given cell-cycle phase. In this review, the contributions of mass spectrometry (MS) to the deciphering of these intricate networked interactions are described along with the numerous applications in which it has proven useful. Such applications include, for example, the identification of the partners involved in NA-P or P-P complexes, the identification of post-translational modifications that (may) regulate such complexes' activities, or even the precise molecular mapping of the interaction sites in the NA-P complex. From a biological standpoint, we felt that it was worth the reader's time to be as informative as possible about the functional significance of the analytical methods reviewed herein. From a technical standpoint, because mass spectrometry without proper sample preparation would serve no purpose, each application described in this review is detailed by duly emphasizing the sample preparation-whenever this step is considered innovative-that led to significant analytical achievements. PMID- 12645092 TI - Pulmonary effects of welding fumes: review of worker and experimental animal studies. AB - BACKGROUND: Approximately one million workers worldwide perform welding as part of their work duties. Electric arc welding processes produce metal fumes and gases which may be harmful to exposed workers. METHODS: This review summarizes human and animals studies which have examined the effect of welding fume exposure on respiratory health. An extensive search of the scientific and occupational health literature was performed, acquiring published articles which examined the effects of welding on all aspects of worker and laboratory animal health. The databases accessed included PubMed, Ovid, NIOSHTIC, and TOXNET. RESULTS: Pulmonary effects observed in full-time welders have included metal fume fever, airway irritation, lung function changes, susceptibility to pulmonary infection, and a possible increase in the incidence of lung cancer. Although limited in most cases, animal studies have tended to support the findings from epidemiologic studies. CONCLUSIONS: Despite the numerous studies on welding fumes, incomplete information still exists regarding the causality and possible underlying mechanisms associated with welding fume inhalation and pulmonary disease. The use of animal models and the ability to control the welding fume exposure in toxicology studies could be utilized in an attempt to develop a better understanding of how welding fumes affect pulmonary health. PMID- 12645093 TI - Health care utilization of families of carpenters with alcohol or substance abuse related diagnoses. AB - BACKGROUND: Patterns of health care utilization of families of carpenters with and without alcohol and/or substance abuse related diagnoses (ASRD) were compared. METHODS: Utilization data for families of 13,657 carpenters for a 10 year period were analyzed. Concordance of diagnoses among family members was assessed and proportionate utilization ratios were used to compare the experiences of families of carpenters with and without ASRD. RESULTS: Twenty eight percent of the spouses with ASRD had a carpenter spouse with a similar diagnosis. Twenty-four percent of the families with a child with ASRD had a parent with one of these diagnoses compared to 9.4% of families without any children with ASRD (P < 0.0001). Families of carpenters with alcohol or ASRD also had different patterns of health care utilization. CONCLUSIONS: Utilization patterns were influenced to a significant degree by concordant diagnoses among spouses and children. Excess mental health care was seen among families of carpenters with ASRD above their care for substance abuse treatment. PMID- 12645091 TI - The effect of cigarette smoking on musculoskeletal-related disability. AB - BACKGROUND: This study describes cigarette smoking's effect on development of physical disability following initial musculoskeletal-related hospitalization. METHODS: We followed 15,140 US Army personnel hospitalized for common musculoskeletal disorders between 1989-1996 for up to 8 years (1997) to assess risk for long-term physical disability. RESULTS: Trends between increased smoking level and long-term disability were identified for persons with knee injuries, rotator cuff injuries, and intervertebral disc displacement. In proportional hazards models, disability was significantly associated with heavy smoking among all subjects (relative hazard (RH) = 1.21). Both heavy smokers (RH = 1.49) and light to moderate smokers (RH = 1.44) were at greater risk for disability following meniscal injuries. Excess fraction due to smoking among subjects with meniscal injuries who currently smoke was 38%. CONCLUSIONS: Findings suggest an association between smoking and development of disability following meniscal injury. Given the high excess fraction of disability associated with smoking, other studies are needed to confirm this association. PMID- 12645094 TI - Occupational risk in health care and research. AB - BACKGROUND: Working in the health care and research sectors has been linked to various hazards. METHODS: Studies published in the peer-reviewed literature that are pertinent to the exposures or diseases relevant to these fields were reviewed. RESULTS: The most important exposures include infectious agents, formaldehyde, anesthetic agents, antineoplastic drugs, and ethylene oxide. The best-documented evidence is that of infectious risk primarily among clinical personnel. Monitoring studies of persons occupationally exposed to anesthetics clearly demonstrate behavioral effects, possible risk of reproductive problems, as well as cytogenetic effects of unknown significance. The latter two impairments are also observed among those exposed to antineoplastic drugs and ethylene oxide. Exposure to formaldehyde appears to be associated with nasopharyngeal tumors. Whereas increased risk of cancer of certain sites, particularly the brain and lymphohematopoietic system, is found among research and health care personnel, no specific exposure has been linked to these neoplasms. CONCLUSIONS: Although some results are inconsistent, continued environmental and biological monitoring will allow better assessment of exposures and of implemented protection measures. PMID- 12645095 TI - Causes of electrical deaths and injuries among construction workers. AB - BACKGROUND: Contact with electrical current is the fourth leading cause of deaths of construction workers. This study evaluates electrical deaths and injuries to construction workers. METHODS: Two sources of data were analyzed in detail: (1) 1,019 electrical deaths identified by the Bureau of Labor Statistics, Census of Fatal Occupational Injuries (CFOI) for the years 1992-1998; and (2) 61 electrical injuries identified between November 1, 1990 and December 31, 1998 from a George Washington University Emergency Department injury surveillance database. RESULTS: Contact with "live" electrical wiring, equipment, and light fixtures was the main cause of electrical deaths and injuries among electrical workers, followed by contact with overhead power lines. Among non-electrical workers, contact with overhead power lines was the major cause of death. Other causes included contact with energized metal objects, machinery, power tools, and portable lights. Arc flash or blast caused 31% of electrical injuries among construction workers, but less than 2% of electrical deaths. CONCLUSION: Adoption of a lockout/tagout standard for construction, and training for non-electrical workers in basic electrical safety would reduce the risk of electrical deaths and injuries in construction. Further research is needed on ways to prevent electrical deaths and injuries while working "live". PMID- 12645096 TI - Occupational asthma symptoms and respiratory function among aerial pesticide applicators. AB - BACKGROUND: Pesticide exposure has been suggested as one causal factor for the rise in asthma prevalence. The goal of this investigation was to determine the effect of pesticide exposure on respiratory symptoms and lung function in workers with occupational exposure to pesticides. METHODS: A prospective, case-controlled study was conducted among pesticide aviators (AV) and community controls (Con). In Phase I, subjects completed an asthma survey and baseline spirometry. In Phase II, subjects reported symptoms, lung function monitoring, and pesticide exposure during two, 14-day periods. RESULTS: Phase I-Self-reported asthma and symptoms were similar among AV (n = 135) and Con (n = 118) with 4-6% prevalence reported but with higher rates among smokers. Baseline lung function was similar; although, a higher proportion of AV had forced expiratory volume in one second (FEV(1)) <80% predicted (8% vs. 2%, P = 0.02). Phase II-Self-reported symptoms were similar with 80% of AV (n = 50) and 73% of Con (n = 49) reporting no symptoms. Only 4% of AV and 6% of controls reported increased symptoms from baseline to spray season. Serial lung function did not differ between group and mean diurnal variation in peak expiratory flow improved in both groups between sampling times (AV 18% vs. 14%; Con 19% vs. 16%, P < 0.001). CONCLUSIONS: This study suggests that among workers with occupational pesticide exposure, asthma symptoms and lung function are similar to those of controls with only community based exposure. PMID- 12645097 TI - Occupational health services at ten U.S. Department of Energy weapons sites. AB - BACKGROUND: Nuclear weapons sites are among thousands of federal facilities that are contaminated with a variety of hazardous wastes. Workers at these sites participate in a wide array of activities that can result in their exposure to a variety of hazards. METHODS: The purpose of this study was to describe the occupational health and safety services at ten U.S. Department of Energy sites. Interviews were conducted with 30 health and safety professionals, including physicians, program managers, and industrial hygienists. Data collection was guided by a quality assessment model that provided a means to examine the structures, processes, and outcomes of occupational health and safety services. RESULTS: It was determined that services for some worker groups, such as subcontractors, were limited; other problems identified included a mismatch between hazards (physical hazards) and types of service providers (radiation experts), ineffective databases, and communication problems. CONCLUSIONS: This description of health and safety services has provided valuable information that can be used to improve the efficiency and effectiveness of service delivery in this large and complex organization. PMID- 12645098 TI - Development and evaluation of an advanced training technology course within a union-based industrial emergency response training program. AB - BACKGROUND: The Health and Safety Department of the United Automobile Workers recently introduced the use of Advanced Training Technologies (ATTs) into a previously successful Industrial Emergency Response (IER) program. The new curriculum attempted to incorporate web-based technology, while preserving the historical emphasis on the use of peer trainers and the Small Group Activity Method (SGAM) to promote team problem solving and worker empowerment. METHODS: The University of Michigan evaluated the program by administering a survey combining open- and closed-ended questions to trainees. RESULTS: Of the 46 respondents, most rated the program very highly, found that the use of internet technology allowed faster access to current information, reported peer trainers as one of program aspects they liked best, and did not find working in groups of three per computer difficult. CONCLUSIONS: It is possible to enhance a workplace health and safety training program through the introduction of ATT without compromising participant-centered, group learning program philosophies. PMID- 12645099 TI - Respiratory symptoms in kitchen workers. AB - INTRODUCTION: A possible association between cooking fumes and respiratory diseases other than cancer has not been studied earlier. METHODS: All employees at 67 selected kitchens were asked to answer a personal questionnaire regarding the presence of dyspnea, serious dyspnea, cough, and respiratory symptoms in connection with work. The study group consisted of 139 women and 100 men. RESULTS: The prevalence of dyspnea (RR = 4.1 (2.7-6.3)), serious dyspnea (RR = 2.9 (1.5-5.7)), and symptoms in connection with work (RR = 4.3 (2.7-6.7)) were statistically significantly higher for the female kitchen workers compared to the controls. For the men only dyspnea (RR = 1.8 (1.4-2.3)) and symptoms in connection with work (RR = 2.1 (1.6-2.7)) showed an increased prevalence. An analysis of possible predictors for respiratory symptoms in connection with work gave an odds ratio of 3.2 (P = 0.000) for "working in a restaurant kitchen." CONCLUSIONS: The results of the study indicate a relationship between working in kitchens and respiratory symptoms. PMID- 12645100 TI - Pulmonary function abnormality and respiratory tract irritation symptoms in epichlorohydrin-exposed workers in Taiwan. AB - BACKGROUND: Epichlorohydrin (ECH) is a strong irritant of the eyes, respiratory tract, and skin. The aims of this study were to examine the dose-response relationship between observed abnormal pulmonary function tests and respiratory tract irritation symptoms among epichlorohydrin-exposed workers in Taiwan. METHODS: A total of 167 workers were randomly selected from a resin synthesis factory. Sixty-six air samples were taken to determine ECH concentration in the workplace. Demographic data, work history, smoking status, and respiratory tract irritation symptoms were gathered by a standard self-administered questionnaire. Pulmonary function tests were also performed. RESULTS: There were 13 of 41 (31.7%) abnormal mean mid-expiratory flow (MMEF) among the high-ECH-exposed workers, 11 of 38 (29%) among the low-ECH-exposed workers, and 4 of 59 (6.8%) among non-ECH-exposed workers. There was a significant linear trend between ECH exposure and the prevalence of small airway abnormalities (P = 0.007) after adjusting for other factors. There was also a significant dose-response relationship of respiratory tract irritation symptoms (cough, phlegm, chest tightness, and dyspnea) among the ECH-exposed workers. CONCLUSIONS: This study suggests that obstructive lung abnormalities and small airway lung damage are associated with ECH exposure. The study also suggests that exposure to very low concentrations (<0.2 ppm) causes significant higher prevalence of respiratory tract irritation symptoms. Causal inferences from the findings cannot be made from this cross-sectional study and further longitudinal studies are needed to better clarify the nature of the observed associations. PMID- 12645101 TI - Apricot sulfurization: an occupation that induces an asthma-like syndrome in agricultural environments. AB - BACKGROUND: Several cases of bronchial asthma have been presenting with acute attacks during the seasons of apricot sulfurization. The aim of this study was to reveal the effects of sulfur dioxide (SO(2)) exposure on the airways of the workers involved in this process. METHODS: SO(2) levels in air were measured on 15 apricot farms, while the symptom scores of 69 workers were recorded before, during, and after SO(2) exposure. Physical examination and pulmonary function tests (PFT) of the workers were also done prior to and after exposure periods. RESULTS: The measured SO(2) concentrations ranged between 106.6 and 721.0 ppm. Dyspnea (80%), cough (78%), and eye and nose irritation (83-70%) were the most commonly observed symptoms. The workers had significant decreases in pulmonary functions after SO(2) exposure. Decrements in FEV(1), FEV(1)/FVC%, and FEF(25 75%) showed that the acute effect of SO(2) on pulmonary functions of the workers was mostly of obstructive pattern. CONCLUSIONS: Acute exposure to SO(2) induces "asthma-like syndrome" (ALS) in apricot sulfurization workers. SO(2) which has not been previously reported in agricultural environments as a cause of ALS may be considered as a new agent. PMID- 12645102 TI - Cause, type, and workers' compensation costs of injury to fire fighters. AB - BACKGROUND: Work-related injury rates in the fire service industry exceed those for most other industries, however little is known about the cost of injury to firefighters. METHODS: This is a preliminary investigation of detailed worker's compensation records of firefighter injuries, 1,343 claims in all, collected from 1992-1999. Summary statistics and regression analysis regarding the cause, nature, and cost of injury to firefighters are presented. RESULTS: Overexertion accounted for a significant portion (over 1/3) of injuries to firefighters, typically involved injuries to the back, and was associated with significantly higher costs than other types of injuries. The per-claim average worker's compensation cost of injury to firefighters was $5,168 and the average for injuries caused by overexertion was $9,715. CONCLUSIONS: Overexertion is a costly source of injury to firefighters that can likely be reduced through policy intervention. PMID- 12645103 TI - [Clinical usefulness of a cardiopulmonary sleep study in patients with sleep apnea syndrome]. AB - Although polysomnography is indispensable in the diagnosis of sleep apnea syndrome (SAS), a common disease in Japan, it is a time-consuming procedure. We therefore assessed the clinical usefulness of a cardiopulmonary sleep study (CPSS), which monitors indices of respiratory and circulatory kinetics during sleep. Of ninety male patients (50 +/- 11 years old, BMI: 27 +/- 4), those with apnea indices (AI) > 5, as determined using an apnomonitor, underwent CPSS with a Night Watch System. Sixty-nine patients with indications for nasal CPAP (NCPAP) therapy underwent overnight CPSS and the optimum pressure was determined. AI was 31 +/- 17, whereas CPSS showed an apnea-hypopnea index of 49 +/- 24, a mean SaO2 of 93 +/- 3%, and a minimum SaO2 of 71 +/- 11%, indicating severe OSAS. The mean optimum pressure was 10 +/- 2 cm H2O, and the compliance was 86%, which was comparatively good. As CPSS can be performed on many patients in a short time, it may be useful for the early diagnosis and initial treatment of SAS. PMID- 12645104 TI - [Chronic eosinophilic pneumonia--a follow-up study of 12 cases]. AB - In this study, twelve cases of CEP were followed for a mean period of 10.9 years (range 9.3-12.9 years) at National Himeji Hospital. Of the five women and seven men examined, two patients possessed preexisting asthma and one developed asthma during the course of CEP disease. None of the patients exhibited any symptoms of allergic rhinitis. All had dramatic responses to corticosteroid therapy without developing extrathoracic manifestations. During the course of CEP, one patient died from acute myocardial infarction. Relapses, which occurred in six patients, responded as well to the treatment as in the original episode. One patient continued long-term oral corticosteroids (5 mg/day) and steroid inhalants (800 micrograms/day) as treatment for asthma. Another was administered steroid inhalants (800 micrograms/day) to treat both asthma and relapsing CEP; two additional patients received 800-600 micrograms/day to prevent relapse. These data indicate that, with proper treatment, the long-term prognosis for patients with CEP is excellent. PMID- 12645105 TI - [Clinical course of pulmonary eosinophilic granuloma--CT evidence of lesion evolution]. AB - We reviewed a clinicopathological study of pulmonary eosinophilic granuloma focusing on the evolution of pulmonary lesions in chest CT scans. Between 1990 and 2002, five patients with pulmonary eosinophilic granuloma were admitted to our hospital. Clinical features, chest radiographs, pathological characteristics and treatments were evaluated. The scans were repeated in four patients at intervals of at least eight months. All patients were men who smoked. The age at onset of the pulmonary disorders ranged from 22 to 45 years, with a mean of 33 years. Most of the initial chest CT scans showed nodular lesions and thin-walled cysts, but the nodular lesions disappeared or were transformed into cystic lesions in the later scans. Most of the final scans revealed thin-walled cysts and emphysematous lesions. The histopathological characteristics of the nodular lesions in the chest CT were correlated with cellular granulomas mainly composed of Langerhans cells and eosinophils, and small granulomas were also seen in the thin fibrous walls of the cysts. All patients quitted smoking and three showed improvement of the pulmonary lesions within three months. PMID- 12645106 TI - [Thromboembolism in lung cancer patients]. AB - We reviewed 649 primary lung cancer patients with special reference to the occurrence of thromboembolism. Thirteen episodes of thromboembolism were detected in 12 (1.8%) of the 649. Eight of the 12 were men, and their mean age was 63. Adenocarcinoma was the predominant cell type. Most patients had an advanced stage of the disease, but in some in the cancer was at an early stage. In 5 cases, the finding of thromboembolisms led to diagnosis of the lung cancer (38.5%). Anticoagulant therapy was performed in 9 cases, of which 8 responded without serious complication. We emphasize the importance of anticoagulant therapy as a therapy indicated for thromboembolism in patients with lung cancer. PMID- 12645107 TI - [Clinical characterization of interleukin-8 in patients with idiopathic pulmonary fibrosis]. AB - The levels of interleukin-8 (IL-8) in the serum, bronchoalveolar fluid (BALF) and epithelial lining fluid (ELF) were measured in patients with idiopathic pulmonary fibrosis. (IPF), in order to evaluate the clinical significance of IL-8. The serum levels were significantly higher in patients with active IPF (34.4 +/- 11.9 pg/ml, n = 8) than in those with stable IPF (mean: 14.6 +/- 10.9 pg/ml, n = 18), but neither correlated with the serum level of KL-6 or of SP-D, or with the intensity of chest Ga67-scintigraphy. There were no significant differences in BALF or ELF IL-8 levels between the active and stable IPF groups. These results suggest that the serum level of IL-8 is a useful marker for evaluating the disease activity in patients with IPF. PMID- 12645109 TI - [Early image of squamous cell carcinoma in the peripheral lung field on high resolution CT scan]. AB - A 65-year-old man presented with the shadow of an abnormal nodule in the left lower lung field in a chest radiograph. We diagnosed this as an old inflammatory change because prior chest radiographs had shown the same nodule in the same lung field. However, a high-resolution CT scan showed a hazy ground-glass opacity (GGO) near the nodule. Two years later, this GGO changed into a small nodule. After a CT-guided transbronchial lung biopsy performed by ultra-thin fiberoptic bronchoscopy, we diagnosed this nodule as squamous cell carcinoma. We speculated that the hazy GGO detected in the peripheral lung field on high-resolution CT two years before diagnosis may have been an early image of squamous cell carcinoma. PMID- 12645108 TI - [Legionella pneumonia: report of 25 community-acquired cases]. AB - We assessed clinical, laboratory and radiologic findings in 25 patients (21 men and 4 women; mean age 60.1 years) with Legionella pneumonia. The diagnosis was confirmed from the isolation of Legionella pneumophila in respiratory samples(nine cases), or by a fourfold increase or decrease in antibody titers in the paired serum samples (16 cases). Twelve patients were smokers, 5 patients were chronic alcoholics and 18 patients had underlying diseases. Regarding severity, the illnesses were mild (three patients), moderate (seven patients) and severe (fifteen patients). The following presenting symptoms and findings that characterize Legionella pneumonia were seen: a temperature of more than 40 degrees C (six patients), disorientation or gate disturbances(ten patients), diarrhea (eight patients), relative bradycardia(eight patients), hyponatremia(five patients), and a high creatinine phosphokinase suggesting rhabdomyolysis(five patients). Rapid progression of infiltrates on chest radiographs before appropriate antibiotic therapy was observed in seven of the twelve patients whose previous chest films were available. Only one patient, who was in endotoxin shock on admission, died. Suspected sources of infection for 13 patients were a hot spring and a public bath. We conclude that a detailed analysis of the above clinical features makes possible the diagnosis of Legionella pneumonia. PMID- 12645110 TI - [A case of hemophagocytic syndrome caused by tuberculosis]. AB - A 59-year-old woman was admitted to our hospital with a picture of high fever and dyspnea. Pneumonia was diagnosed by chest CT scan and was treated. Laboratory findings showed pancytopenia and examination of aspirated bone marrow showed mature histiocytes with marked hemophagocytosis. She was treated with prednisolone, but her respiratory condition worsened, so she was ventilated. An acid-fast bacterial smear of bronchoalveolar lavage fluid from the left lingula showed Gaffky's scale No. 5, and so tuberculosis was diagnosed. Though anti tuberculosis therapy was started, she died after eight days of hospitalization. Infection-associated hemophagocytic syndrome is caused by various infections. Until now, 26 cases of tuberculosis-associated hemophagocytic syndrome have been reported, and many of these diseases were caused by severe tuberculosis. If a patient with severe tuberculosis presents with leucocytopenia or thrombocytopenia, investigations should serve a differential diagnosis that takes into account disseminated intravascular coagulation, tuberculous granuloma of the bone marrow, side effects of anti-tuberculous drugs, and hypersplenism from hemophagocytic syndrome. PMID- 12645111 TI - [A case of adult-onset Still's disease complicated with acute respiratory distress syndrome]. AB - A 24-year-old woman was admitted to our hospital because of a high fever that had persisted for two weeks. She complained of a sore throat and arthralgia, and had evanescent rash, lymphadenopathy, liver dysfunction, and hyperferritinemia. Tests for RF and ANA were negative. Adult-onset Still's disease was diagnosed. On the fifth day of hospitalization, acute respiratory distress syndrome (ARDS) and disseminated intravascular coagulation (DIC) developed. Treatment consisted of mechanical ventilation and administration of steroid pulse-therapy and gabexate mesilate. Analysis of fluid obtained by bronchoalveolar lavage showed increases in the total cell count, predominantly of neutrophils and lymphocytes. Bilateral pulmonary infiltration seen on chest radiographs was alleviated, and the arterial blood gas data gradually improved. After cyclosporine was given, all the above symptoms associated with adult-onset Still's disease disappeared. Plasma levels of inflammatory cytokines decreased with the improvement of the patient's clinical condition. PMID- 12645112 TI - [A case of pulmonary arteriovenous malformation treated by transcatheter embolization using coils]. AB - The patient was a 21-year-old woman who, at 16 years old, had had no subjective symptoms. She was referred to our hospital because of abnormal shadows in the middle of the left lung that were seen on a chest radiograph during a school medical screening. In her first examination at our hospital, CT revealed tumor like shadows with diameters of 2.0 cm in Lt-S6 and 0.5 cm in Rt-S5, together with arteries and veins. Lung ventilation and blood flow scintigraphy was used to obtain images of both kidneys and the head, and as a result, a pulmonary arteriovenous malformation (PAVM) was diagnosed. Because of a low shunt fraction of 10.8%, the patient was only kept under observation. However, at 21 years of age, she complained of dyspnea with exercise and was admitted to the hospital for further examination. Chest CT showed an increase in the diameter of the arteriovenous malformation (AVM) to 2.7 cm, and 3-D image reconstruction clearly revealed the position of the tumor in relation to the pulmonary artery and vein. The shunt fraction had increased to 18.3%. Coil embolization was therefore performed to A6b, the artery feeding the AVM (using interlocking detachable coils of 6 mm-10 cm and 8 mm-20 cm, and Trufill coils of two 7 mm-6 cm and 5 mm-4 cm). The patient had no serious postoperative complications. A decreased AVM diameter of 1.7 cm and an improved shunt fraction of 11% were observed, and she was discharged from the hospital. Since then, she has been well, without dyspnea, and her chest films show only coils and no aggravation of other AVMs. AVMs generally have no symptoms, but they can cause severe complications such as hypoxia from a right-to-left shunt, hemoptysis, hemothorax, and brain abscesses. Coil embolization should be considered more often for the treatment of AVM because it is safer, more effective and less invasive than traditional operations. PMID- 12645113 TI - [A case of AIDS-associated Haemophilus influenzae pneumonia with diffuse reticulonodular shadows]. AB - A 32-year-old male was admitted to our hospital complaining of fever and dyspnea on effort. Laboratory data on admission indicated leukocytosis and elevation of C reactive protein. A chest radiograph showed diffuse reticulonodular shadows in both lower lung fields, and a chest computed tomography showed centrilobular reticulonodular opacity. Bronchoscopic findings revealed a large amount of slightly yellowish secretion in all bronchi. Cells found in the bronchoalveolar lavage fluid (BALF) included 61% neutrophils. Haemophilus influenzae was isolated from cultures of the BALF and sputum. Transtracheal lung biopsy specimens showed focal infiltration of neutrophils in the alveoli, and the pathological findings in the lung were compatible with bronchiolopneumonia. Since the CD4/CD8 ratio was 0.09 and a positive reaction was obtained for anti-human immunodeficiency virus (HIV) antibody, HIV-associated pneumonia due to H. influenzae was diagnosed. Seven days' administration of cefozopran improved the patient's condition. It is interesting that radiological findings are often unusual in HIV-infected patients with H. influenzae pneumonia. PMID- 12645114 TI - [An autopsy case of cor pulmonale due to a pulmonary tumor embolism as the first clinical manifestation of occult gastric cancer]. AB - A 47-year-old man was admitted to our hospital because of progressive dyspnea and cough. Physical examination and chest radiographs showed the signs of cor pulmonale. A lung scan using perfused radionuclide revealed multiple peripheral perfusion defects and catheterization of the right heart showed severe pulmonary hypertension. A diagnosis of severe pulmonary embolism was made. Despite intensive care with anti-coagulation therapy, the patient died on the third hospital day. Autopsy disclosed gastric cancer in the pylorus with metastases to the regional lymph nodes. There were no macroscopic pulmonary artery emboli or parenchymal lesions, but more than 60% of the small arteries and arterioles were occluded by casts of tumor cells. Cor pulmonale due to a pulmonary tumor embolism is a rare complication of cancer. This case is particularly unusual because the embolus-caused cor pulmonale was the initial manifestation of clinically occult, but pathologically advanced, gastric cancer. PMID- 12645116 TI - [Diagnosis of ischemic heart disease by imaging techniques; cardiologist's perspective]. PMID- 12645115 TI - [A case of hemoptysis caused by vegetable foreign body (cryptomeria) and actinomycosis]. AB - A 54-year-old man was admitted to our hospital with hemoptysis. Chest radiography and chest CT scanning demonstrated atelectasis in the right middle lobe. Bronchoscopy showed nothing of abnormal appearance. We performed a middle lobe lobectomy suspecting that the continuing hemoptysis was caused by the lesion in the middle lobe. Histologically, a vegetable foreign body (cryptomeria) was recognized in a bronchiole of the middle lobe, surrounded by inflamed tissues and sulfur granules. It was suggested that all of these were the cause of the hematoptysis. The patient was discharged on the fourteenth postoperative day, and has been asymptomatic since. This was a very rare case of hemoptysis caused by a vegetable foreign body and actinomycosis. PMID- 12645117 TI - [Diagnosis of ischemic heart disease by ultrasound]. PMID- 12645118 TI - [Nuclear cardiology: diagnosis of myocardial perfusion, metabolism and function]. PMID- 12645120 TI - [Cardiac imaging using multi-detector-row CT (MDCT)]. PMID- 12645119 TI - [Cardiac MRI: current status and future prospective]. PMID- 12645121 TI - [Clinical evaluation of gastric diverticulum on indirect radiograph for gastric mass survey]. AB - The aim of this study was to evaluate the clinical conditions of gastric diverticulum. Fifty-four patients with gastric diverticulum (20 men and 34 women among 34,314 patients who underwent medical check-ups) were evaluated on indirect radiographs, for an incidence of 0.16% among the total number of examined cases, a rate lower than that of previous reports. Almost all cases were asymptomatic, had a single diverticulum, and showed a saccular shape. The age distribution indicated higher frequencies in the 5th and 6th decades, and the posterior wall of the fornix was the most common location. Size ranged from 0.6 cm to 12 cm, and 41 cases (75.9%) were between 1.0 cm and 4.0 cm in size. This entity should be kept in mind when reading radiographs of upper gastrointestinal series as well as recognition of pseudodiverticulum and aberrant pancreas as noted for the stomach in several past report. Diverticulum on the cardia, which was previously classified as gastric diverticulum, should be excluded because of the possibility of normal variation. PMID- 12645123 TI - [High-dose-rate brachytherapy with local injection of bleomycin for N0 oral tongue cancer--possibilities of the control of tumor implant by inserting applicators and the decrease in tumor dose]. AB - Twenty-eight patients with N0 oral tongue cancer were treated with high-dose-rate (HDR) interstitial brachytherapy combined with local injection of bleomycin between December 1997 and June 2001 at the Department of Radiology, National Kyushu Medical Center Hospital. A median dose of 5 mg of bleomycin was injected locally, and 16-20 Gy was delivered to the area surrounding applicators for control of the tumor implant during the initial two days. The two-year local recurrence-free survival rate was 96% [T1, 2: 100% (8/8, 15/15), T3: 80% (4/5)]. The two-year secondary neck node metastasis rate was 7.1% [T1: 12.5% (1/8), T2: 6.7% (1/15), T3:0% (0/5)]. There were no tumor implants in any patients. We tried to decrease the minimal tumor dose step by step. The groups with median minimal tumor doses of 60 Gy, 50 Gy, and 40 Gy had local recurrence rates of 12.5% (1/8), 0% (0/14), and 0% (0/6), respectively. Local recurrence rates were not increased by decreasing the minimal tumor dose. Two patients (7%) had secondary neck node metastasis. Late adverse effects were tongue ulcer: 11% (3/28), oral floor ulcer: 4% (1/28), and osteonecrosis: 4% (1/28). These results suggest that control of the tumor implant and the decrease in minimal tumor dose below 60 Gy may be possible with the local injection of bleomycin and delivery of doses to the area surrounding the applicators when NO tongue cancer is treated using 192Ir-HDR brachytherapy. PMID- 12645122 TI - [Impact of treatment extending to 60 Gy on the outcome of radiotherapy for nasopharyngeal carcinoma]. AB - PURPOSE: To clarify the impact of treatment duration on the outcome of nasopharyngeal carcinoma (NPC). MATERIALS AND METHODS: Forty-three patients with NPC were treated with definitive radiotherapy from January 1980 through May 1996. The male-to-female ratio was 32:11, and median age was 58 years (10-78 years). According to the fifth UICC classification, 4 patients were stage I, 12 were stage II, 6 were stage III, and 21 were stage IV. Twenty-nine patients received chemotherapy. Each patient was treated to various doses according to their disease extension. Thus, treatment duration was defined as the duration from the start of radiotherapy to the end of 60 Gy. The median follow-up period was 63 months (2-164 months). RESULTS: The 5-year overall and disease-free survival rates of all patients were 66% and 59%, respectively. The 5-year disease-free survival rates of the patients treated with the short treatment duration (< or = 8 weeks) and those treated with the long treatment duration (> 8 weeks) were 76% and 38%, respectively (p = 0.008). CONCLUSION: Long treatment duration may lead to poor treatment outcome in NPC. PMID- 12645124 TI - [Simple visualization of the corticospinal pathway using tractography: one-ROI and two-ROI methods]. AB - Diffusion tensor imaging (DTI), a magnetic resonance (MR) technique to analyze diffusion anisotropy of the brain, is able to demonstrate subtle white matter anatomy. Tractography is expected to be a unique, non-invasive tool to provide more pertinent insights into brain structure and orientation not accessible by conventional MRI. In this report, preliminary experiences of visualization of the corticospinal tract using tractography are described. DTI of the brain was performed in 5 normal volunteers using single-shot echo-planar imaging, then tractography was generated by our original software. We determined that the two region-of-interest (ROI) method is superior to the one-ROI method. PMID- 12645125 TI - [Evaluation of the anisotropy in the patients with Alzheimer's disease]. AB - Diffusion-weighted MR imaging was performed in 5 control subjects and 8 patients with Alzheimer's disease(AD). The mean values of fractional anisotropy(FA) were calculated in the genu and splenium of the corpus callosum, and anterior and posterior cerebral white matter. The FA value of the posterior white matter in patients with AD was significantly lower than that of controls. Fractional anisotropy may be useful in the diagnosis of AD. PMID- 12645126 TI - [Imaging findings of elongated styloid process syndrome (Eagle's syndrome): report of two cases]. AB - Elongated styloid process syndrome, which is synonymous with Eagle's syndrome, is a rare disease. Although an elongated styloid process is not always symptomatic, if it compresses the cranial nerves and/or carotid artery, a patient can present with sore throat, dysphasia, or dysphoria of the throat. We present two cases of elongated styloid process syndrome, focusing on the imaging findings. The elongated styloid processes were visualized by panoramic photograms. However, CT images, especially 3DCT images, were useful in visualizing the elongated styloid process itself as well as the relationship between the styloid process and surrounding soft tissue. PMID- 12645128 TI - [E-Cell simulation system and its application to the modeling of circadian rhythm]. PMID- 12645127 TI - [CT-guided radiofrequency ablation of renal cell carcinoma in a patient with solitary kidney after contralateral nephrectomy: a case report]. AB - A 62-year-old man with renal cell carcinoma in the solitary kidney after right nephrectomy underwent CT-guided radiofrequency ablation. The tumor, which was located in the dorsal part of the lower aspect of the kidney, was exophytic and measured 2 cm in diameter. A 17-gauge, internally cooled radiofrequency needle electrode was placed percutaneously into the center of the lesion. Dynamic CT 6 days after the first ablation showed band-like residual lesion along the intact renal parenchyma. A second ablation was performed for the residual lesion. CT both 1 and 3 months after the second procedure demonstrated complete ablation with the normal renal parenchyma maintained. PMID- 12645129 TI - [Lipoylation of the proteins having lipoic acid as a prosthetic group]. PMID- 12645130 TI - [Function of zfhx1 family and molecular genetic analysis of its deficiency]. PMID- 12645131 TI - [Advances in the Maillard reaction and glycation researches--mainly on the Namiki pathway]. PMID- 12645132 TI - [A molecular programme for the germ line specification in mice]. PMID- 12645133 TI - [Retrograde synaptic transmission in invertebrate model organisms]. PMID- 12645134 TI - [Visualizing exocytosis of insulin granules with evanescent wave microscopy]. PMID- 12645135 TI - [Recent progress in protein folding research]. PMID- 12645136 TI - [Macrophage responses during phagocytosis of apoptotic cells and their regulatory mechanism]. PMID- 12645137 TI - [A novel and practical synthesis of (+)-biotin via Fukuyama coupling reaction]. AB - (+)-Biotin (1) was synthesized from readily accessible L-aspartic acid (4). The contiguous asymmetric centers at C-3a and C-6a were formed through a diastereoselective aldol reaction of N-Cbz-3-amino-4-butanolide 5 to provide trans-disubstituted lactone 6 with high stereoselectivity (trans/cis = 12:1). The imidazolidin-2-one moiety of 1 was constructed by a stereoselective Hofmann rearrangement of beta-substituted asparagine derivative 7 to provide cyclic urea 8. This reaction proceeds with complete retention of stereochemistry. Removal of the protective groups of 8 and subsequent dibenzylation and thionation provided thiolactone 2. The installation of the C-4 side chain of 1 was performed through a Pd/C-catalyzed coupling reaction of 2 with ethoxycarbonylbutylzinc iodide 14a (Fukuyama coupling reaction), which permitted the synthesis of 1 from 2 under industrially applicable mild conditions in three steps. PMID- 12645138 TI - [Studies on the marker compounds for standardization of traditional Chinese medicine "polyporus sclerotium ([symbol: see text])"]. AB - Natural products have been used for healthcare and pharmacotherapy. Because difficulties in quality control affect their production, processing, and marketing, it is necessary to establish adequate marker compounds for their effective application. Ergosta-4,6,8(14),22-tetraen-3-one(I) was studied in the screening of the marker compounds for the standardization of Polyporus Sclerotium ([symbol: see text]), which has the advantage of easy qualitative and quantitative analysis because of its fluorescence. Its applicability in the standardization of Polyporus Sclerotium is discussed based on comparative studies of 30 crude samples of Polyporus Sclerotium and some other fungi herbs using TLC and HPLC analysis with I it as the marker compound, as well as its chemical synthesis. PMID- 12645140 TI - [Creation and usefulness of the "drug information card" for support of dispensing]. AB - We placed a "drug information card" on each medication shelf to assist dispensing by pharmacists, education of students, and risk management of pharmaceutical practices at the Department of Pharmacy, University of Tokyo Hospital. To provide appropriate information items on the drug information cards, we conducted questionnaire survey of 41 pharmacists on the utility of the cards and reviewed questions received from medical staff in the drug information section in our hospital. Based on the results of these investigations, "usage and dosage," "interactions," "contraindications," "product name," and "effects" were printed as basic information items on the drug information cards. Furthermore, information on pharmacokinetics was added. To make maintenance easier, we classified drug information items into "renew often" and "not so often." A good response on the use of the drug information cards was received for dispensing support from 38 pharmacists (92.7%), 14 trainees (100%), and 16 students (94.1%) in the questionnaire investigation. The drug information cards make it possible to obtain precise information rapidly in pharmacies. PMID- 12645139 TI - [Clinical usefulness of serum total cholesterol as an index of hypothyroidism in patients after cervical radiation]. AB - Cervical radiation therapy is often applied to patients with head and neck cancers because radiation has a high sensitivity to these cancers and permits the preservation of functions and physical form. However, it has been shown that various complications can result from radiation therapy. We have encountered some patients who showed hypercholesterolemia resulting from cervical radiation. Therefore, we have paid close attention to the relationship between hypercholesterolemia after cervical radiation and hypothyroidism. Thyroid hormone tests in these patients with hypercholesterolemia after cervical radiation showed high thyroid stimulating hormone (TSH) and low free thyroxine (fT4), indicating the presence of hypothyroidism. After administration of levothyroxine Na, their fT4 levels increased and both TSH levels and serum total cholesterol levels decreased. In conclusion, in patients who have received cervical radiation, we recommend monitoring serum total cholesterol periodically to detect hypothyroidism easily before the appearance of its symptoms. PMID- 12645141 TI - [A necessity of control of the medical doctor for health center's head with reading the interim report of the council for decentrization reform]. PMID- 12645142 TI - [Mental health and psychosocial factors with single-child high school students in an urban city of China]. AB - OBJECTIVE: This study investigated psychosocial factors underlying the mental health problems of single-child high school students in China, where society and the family situation have been rapidly changing since introduction of the open economy policy. METHOD: Three hundred and ten college-bound high school students in Heilong Jiangsheng Harbin completed self-administrative questionnaires in February, 2000. The subjects were divided into single-child and non single-child groups. Analysis of correlations was performed for general attributes, mental conditions measured by General Health Questionnaire (GHQ), personality variables, stressors, and emotion support network. Cause-and-effect factors were also analyzed using Covariance Analysis. RESULT: In the single-child and in non single child groups, the percentage suffering neurotic tendencies were 73% and 39%, and the values for a tendency to depression were 63% and 25%, respectively. In the single-child group, anxiety, interpersonal dependence, and perceived stressors were significantly higher while the perceived self-esteem and emotional support from family members were significantly lower than in the non single-child group. Among the variables, having siblings was highly correlated with all the measured factors influencing mental health. The results indicated that a poor emotional support network could cause low self-esteem, high anxiety trait, strong interpersonal dependence, and increased sensitivity to stressors and worsening of mental health. CONCLUSION: The incidence of mental health related problems was found to be significantly higher in the single-children than in the non single children. Thus having siblings has positive effects on mental health. The emotional support network also plays an important role in the mental condition, development of a healthy personality, and building a positive attitude toward stressors. PMID- 12645143 TI - [An analysis of the demand for influenza vaccination among the elderly in Japan]. AB - PURPOSE: This paper will discuss an analysis of the demand for vaccination for the elderly as a high-risk group. Influence of the governmental endorsement and/or subsidy on these demand is, then, evaluated from the estimation results. METHODS: Original data were obtained from two surveys conducted by the author for the elderly living with and without descendents. Information was collected about the elderly themselves, the household, experience of influenza and immunization in the last season, and the hypothetical questionnaire about immunization was answered by each respondent to be applied for Conjoint Analysis. Three estimations are performed for the actual behavior, Conjoint Analysis and the joint estimation of these two methods. RESULTS: Experience of influenza and immunization in the last season, proved to be two of the most important determinants. Among the others estimated parameters, cost of immunization, the number of immunization to complete for effectiveness, availability of the immunization at night or on a weekend, and the governmental endorsement greatly affected the immunization demand. Moreover, the superiority of the statistical properties of the joint estimation was confirmed. CONCLUSIONS: The estimation results imply that about 8.9 million elderly people would demand vaccination if there was no cost and there was a governmental endorsement. This would be reduced to be 3.2 million if the cost was 6,000 yen (about 50 dollars) and there was no governmental endorsement. Governmental endorsement alone would increase the number by 2.0 million. The change from no cost to only 500 yen (about 4 dollars) would depress the demand by 1.6 million. PMID- 12645144 TI - [Comprehensive health examination ("Otasha-Kenshin") for the prevention of geriatric syndromes and a bed-ridden state in the community elderly. 1. Differences in characteristics between participants and non-participants]. AB - PURPOSE: We conducted a comprehensive mass health examination for elderly subjects aged 70 or above, living in the community, to assess early deterioration to long-term care conditions and promote healthy and successful aging ("Otasha Kenshin"). In this study, we clarified characteristic differences between participants and non-participants. SUBJECTS & METHODS: A mass health examination was offered in October 2001 to 863 community elderly, including individuals suffering from falls (and fractures), incontinence, malnutrition, depression, mild cognitive impairment and less of functional capacity. Among the total, 438 (50.8%) opted for the "Otasha-Kenshin" examination. Differences in characteristics between the participants and non-participants were examined, parameters including sex and age distribution, self-rated health, functional capacity by the TMIG Index of Competence, depressed status by the General Health Questionnaire (GHQ), subjective well-being by the Philadelphia Geriatric Center Morale Scale: A revision (PGC-MS), frequency of falls, and prevalence of chronic diseases. The comparison was based on the results of measurements from the final survey conducted in 2000. RESULTS: 1) The participation rate in the "Otasha Kenshin" was 49.0% in males and 51.0% in females. The average age was 75.3 year olds in participants and 76.4 in non-participants, the difference being significant (t = 3.97, P < 0.0001). 2) Non-participants had a significantly lower level of self-rated health than participants. 3) There was no significant difference in hand grip strength between participants and non-participants. 4) Non-participants showed significantly lower level of functional capacity and subjective well-being, and they were more likely to be in a depressed state than participants. 5) There was no significant difference in fall rate between participants and non-participants. 6) The prevalence of hypertension and diabetes (found in more than 5% among the subjects surveyed in 2000) was not significantly different between participants and non-participants. CONCLUSION: With aging of society, new and specialized health maintenance systems for the elderly are essential, both for the prevention of deterioration to a long-term care condition (a bed-ridden status) and for the promotion of successful aging with autonomy. Participants in "Otasha-Kenshin" appear to be healthier and more independent than non-participants who were more frail and at higher risk group of a long-term care condition and a bed-ridden status. The major reason for non-participation in the health examination found in this study was subjective or mental deterioration rather than the presence of chronic illness or any geriatric syndrome per se. Frail elderly people like the non-participants in this study should be encouraged and mentally supported to avoid aggravation of their health status through intensive or specialized health surveillance system such as home-visit nursing. PMID- 12645145 TI - [Factors associated with participation in medical checkups of the elderly at home comparison of 3 regions with different social backgrounds]. AB - PURPOSE: Annual medical checkups are presumed to be important with the well-being of aged individuals. The purpose of this study was to investigate factors related to participation in medical checkups of elderly persons at home. METHODS: A survey was performed targeting so-called young elderly persons living at home in 3 regions in Hokkaido Prefecture, a large city (Sapporo), an ex-coalmining town (Yubari) and a small farming town (Takasu). The study populations were asked about medical checkups, economic status, self-rated health status, activities of daily living (ADL) and basic characteristics such as age, sex and occupation. We used the Mantel-Haenszel method for statistical analysis. RESULTS: 1) Elderly males who received medical checkups within the last 1 year (participants) had a shorter period of education, were less likely to have or to have had the smoking habit, undertook more physical activity, had better eating habit, more often rated their own health as good or normal, and had a better physical ADL & instrumental ADL, than those who had never received medical checkups (non participants). Regionally, the items that showed significant differences included: "live with the spouse", "do not smoke" and "do not have physical pain or disabilities" in Sapporo; "enjoy physical activity", "have good eating habit" and "have good instrumental ADL" in Yubari; and "engaged in agriculture/fishery work" and "had outpatient visits in the last three months" in Takasu. 2) Elderly female participants who had outpatient visits in the last three months had anxiety about their own health, significantly different from non-participants. Regionally, the items that showed significant differences included: "often drink alcohol" in Sapporo; "still working right now", "was engaged in agriculture/fishery work in the past" and "had outpatient visits in the last three months" in Yubari; and "have own house", "do not smoke" and "pay attention to eating habit" in Takasu. 3) The item which showed a significantly high rate both for male and female participants was "still working right now". CONCLUSIONS: This study indicates that medical checkups should be recommended to the elderly, taking into account not only their basic characteristics such as sex and occupation, but also lifestyle, self-rated health status and ADL, which are associated with medical checkup participation. PMID- 12645146 TI - [Smoking scenes in Japanese television dramas]. AB - BACKGROUND: Exposure to smoking scenes in movies and TV dramas has been known to be a trigger for young people starting habitual smoking, but it has not been clarified to what extent youth is routinely exposed to smoking scenes in television programs in Japan. OBJECTIVE: To clarify the status quo of smoking scenes and smoking-related items in serial TV dramas targeting young audience in Japan. METHOD: Seven TV dramas targeting a young audience broadcast between July and September, 2001 were analyzed. A total of 63 hours of TV programs were divided into units of three minutes (a total of 1,264 units). All the units were reviewed for smoking scenes, description of social contexts related to smoking behavior, existence of smoking and smoking-related items. FINDINGS: Among the 1,264 three-minute units, 129 (10.2%) contained smoking behavior, and 258 (20.4%) depicted smoking environment with presence of smoking-related items. Smokers were male actors (126 units, 97.7%), aged 20-40 years (118 units, 91.5%), leading actors (72 units, 55.8%), and were smoking alone (80 units, 62.0%). Smoking places mainly took place in indoor settings (56 units, 43.4%). Ashtrays were the most frequently observed smoking-related items (8.1% of the total units). The smoking scenes with actions other than smoking itself accounted for 70 units (54.3% of the smoking scenes). As for the actions other than smoking, working (28 units, 21.7%) and eating (17 units, 13.2%) were commonly observed. Regarding smoking scenes without any other actions, 32 units (24.8%) were observed in the beginning or ending telops, and 27 units (20.9%) showed a person smoking without doing anything else. There were only 3 units (0.2%) which were against smoking behavior. Approximately 30% of the Japanese serial television drama scenes featured tobacco use in story plots. As the general features of the smoking scenes, most of them do not necessarily require smoking in the story. Furthermore, there was little consideration about separation of designated smoking areas. PMID- 12645147 TI - Optimal temperature of continuous lidocaine perfusion for the heart preservation. AB - OBJECTIVE: During cardiovascular surgery, lidocaine is administered to the cardioplegic system to stabilize cell membranes and prevent arrhythmia. Lidocaine is also commonly used in hypothermia. Both lidocaine and hypothermia are myocardially protective. Under normothermia, lidocaine displays its full pharmacological effects, which are apt, however, to be suppressed under hypothermia. We conducted experiments to determine the optimal temperature for myocardial protection in continuous lidocaine cardioplegia. METHODS: In Langendorff mode, rat hearts were continuously perfused with 1 mMol/l of lidocaine solution at 36 +/- 0.5 degrees C (Group A), 24 +/- 0.5 degrees C (Group B), or 7 +/- 0.5 degrees C (Group C) during preservation. Cardiac function and intracellular calcium concentration were measured during both preservation and reperfusion. Heat shock protein 70 (HSP70) was subsequently analyzed by Western blotting. RESULTS: Rapid cardiac arrest was obtained in Groups A and C. Heart rate recovery was good and ultimately the best in Group B, but worst in Group A. During lidocaine perfusion, the volume of coronary perfusion flow decreased gradually in all groups. After reperfusion, Group A showed only a slight increase in coronary perfusion, While Groups B and C showed a marked increase. Left ventricular contractility showed good recovery in all groups. The calcium concentration increased slightly in Group A, but decreased in Groups B and C. No calcium overload was evident in Group A. The same HSP70 level was detected in all groups. CONCLUSION: Lidocaine used in normothermia does not decrease cardiac metabolism or oxygen consumption, and displays full pharmacological effectiveness in preventing ischemic injury. We found 36 degrees C to be the optimal temperature for heart preservation by coronary perfusion with lidocaine cardioplegia. PMID- 12645148 TI - Early results of complete off-pump coronary revascularization using left internal thoracic artery with composite radial artery. AB - OBJECTIVE: Off-pump coronary artery bypass (OPCAB) using the left internal thoracic artery (LITA) with a composite radial artery (RA) was employed for arterial revascularization in order to minimize neurological complication. METHODS: Sixty-one patients underwent OPCAB using the LITA with a composite RA. Angiography was performed in all patients at two weeks postoperatively. RESULTS: The mean number of distal anastomoses was 3.2 +/- 0.4. A Y-composite graft was used in 55 patients, and K-composite graft was used in the other 6 patients. There was no hospital death, no neurological complication nor deep sternal infection. Furthermore, there was no episode of perioperative myocardial infarction nor hypoperfusion syndrome. Patients have been angina-free during a mean follow-up period of 1 year. The graft patency of the LITA to the left anterior descending artery (LAD) was 100% (61/61 anastomoses). The RA became occluded in 4 patients, and the patency rate was 95.6% (130/136 anastomoses). String or coronary-coronary bypass resulting from flow competition was observed in the LITA of 6 patients and in the RA of 13 patients. The string of the LITA occurred in the segment distal from the anastomosis with the composite RA. The string or coronary-coronary bypass was observed more often in cases in which the recipient coronary artery had less than 75% stenosis. CONCLUSION: OPCAB using only the LITA with a composite RA can be successfully and safely performed in patients with multivessel disease. Late postoperative follow-up of the flow competition is necessary to delineate the significance of flow competition. PMID- 12645149 TI - Salmonella pericarditis in a patient with primary idiopathic chylopericardium. AB - We report a case of a 40-year-old man with chylopericardium who developed purulent pericarditis caused by Salmonella enteritidis. Thoracoscopic pericardiotomy with debridement effectively controlled the pericardial infection. PMID- 12645150 TI - Successful repair of left ventricular pseudoaneurysm after mitral reoperation under hypothermic circulatory arrest. AB - Successful repair of left ventricular (LV) pseudoaneurysm after mitral valve replacement (MVR) is uncommon, with delayed onset showing particularly high mortality. We report a case of delayed LV pseudoaneurysm onset after MVR reoperation. The patient had unique clinical symptoms, including coronary ischemia due to compression of the left main coronary artery by the aneurysm. Surgical repair was done using hypothermic circulatory arrest via left thoracotomy. The patient has experienced no cardiac events or pseudoaneurysm recurrence in the 7 years since surgery. PMID- 12645151 TI - Lung cancer associated with sarcoidosis. AB - Sarcoidosis is often identified as swollen hilar lymph nodes found in chest radiography during routine physical checkups. We report a patient with concomitant sarcoidosis and lung cancer necessitating thoracoscopic lymph node biopsy to differentiate between sarcoidosis and lung cancer as the cause of mediastinal/hilar lymph node swelling prior to conducting pulmonary lobectomy. Thoracoscopic biopsy of left mediastinal lymph nodes and sampling of frozen sections of right lymph nodes during intraoperative diagnosis did not detect metastasis. A permanent pathological slide, however, indicated that right interlobar (#11s) lymph nodes involved both sarcoidosis and lung cancer metastasis. Despite careful preparation, it thus remains difficult to distinguish between these conditions even when lymph nodes are strictly evaluated. PMID- 12645152 TI - Biventricular repair for pulmonary atresia with intact ventricular septum associated with sinusoidal communications. AB - Pulmonary atresia with intact ventricular septum (PA-IVS) is very difficult to treat due to the variety of right ventricular hypoplasia and coronary artery anomalies. Biventricular repair is viewed as possible in patients with mild right ventricular hypoplasia but not in those with severe right ventricular hypoplasia or with sinusoidal communication. We report a case of a 17-day-old boy with PA IVS and severe right ventricular hypoplasia and large sinusoidal communications. We successfully conducted biventricular repair, ligated coronary artery and right ventricular fistulas, and implemented right ventricular outflow tract reconstruction. He now enjoys good health and attends elementary school. Sinusoidal communications are not always a limiting factor in biventricular repair for PA-IVS if the right ventricle can be decompressed without inviting ventricular dysfunction by ligating fistulas. PMID- 12645154 TI - Rupture of a small fusiform thoracic aortic aneurysm. AB - We report a case of ruptured aortic aneurysm of the distal aortic arch. This aneurysm was fusiform and only 4.0 cm in maximum diameter. Emergency total aortic arch replacement was successful. Rupture of a thoracic aortic aneurysm of this shape and size is possible but extremely rare. PMID- 12645153 TI - Thymic carcinoid tumor. AB - In this report on 3 cases of thymic carcinoid, tumors recurred in 2 patients not undergoing lymph node dissection. We review 47 cases of this disease taken from the Japanese literature, and discuss surgical procedures for reducing recurrence. In 9 of 20 cases of recurrence in encapsulated thymic carcinoid, lymph nodes were not dissected in eight. Differential diagnosis of an anterior mediastinal tumor should include thymic carcinoid, for which lymph nodes should be routinely dissected, even when the tumor is encapsulated. PMID- 12645155 TI - Bronchogenic cyst with high serum carcinoembryonic antigen. AB - A 64-year-old woman had a large subcarinal cyst accompanied by high serum carcinoembryonic antigen (CEA) of 55.7 ng/ml removed thoracoscopically. The pathological diagnosis was benign bronchogenic cyst. Immunohistochemical findings clearly showed CEA in the stratified columnar epithelial lining of the cyst. Serum CEA returned to the normal range within 40 days of surgical resection. PMID- 12645156 TI - Open retroperitoneal irrigation for graft infection after thoracoabdominal aortic aneurysm repair. AB - A successfully-treated case of graft infection after thoracoabdominal aortic repair is reported. A 61-year-old male underwent graft replacement of a Crawford type IV thoracoabdominal aortic aneurysm and developed graft infection due to methicillin-resistant Staphylococcus aureus. After 35 days' open retroperitoneal irrigation, the culture around the graft turned negative, and omental transposition and skin flap transfer were successfully conducted. The long-term open retroperitoneal irrigation could be conducted without significant complications, and it was very effective in controlling the severe infection. PMID- 12645157 TI - The venous drainage of the human myocardium. AB - New cardiological techniques such as coronary sinus catheterization and selective catheterization of the cardiac veins permit the opening of new experimental and clinical fields, for instance in venous angiography and the reverse nourishment of myocardium which is endangered by ischemia,and also in the electrophysiological study of the components of the conduction system. New approaches in heart surgery, such as the removal of accessory pathways of the conduction system (as in WPW syndrome), necessitate the realization of the topographical relationships of the vessels in the various sections of the coronary sulci in a different way. The objective of this work is, therefore, to present comprehensive and almost new macro- and microanatomical data about the venous drainage of the myocardium via the coronary sinus and its related and unrelated (non-coronary) cardiac veins. Examination of meticulously dissected heart specimens (of individuals who had achieved old or extreme old age at the time of their death in Germany: n=250) as well as corrosion casts of adult cardiac vessels (of individuals of all ages, n=25) formed the basis for the exact description and documentation of the occurrence, frequency, origin, and courses of both the normal and anomalously developed human coronary sinus and cardiac veins. A wide range of morphological and experimental references was consulted in order to enable thorough discussion of the anatomical findings in the light of modern cardiological diagnostics and treatment. The anatomical and clinical nomenclature is presented and there is a brief comment on modern diagnostic techniques and their applications where the cardiac veins are concerned. The two principal and one compound cardiac venous system are defined and discussed with reference to the existence of both the normal and anomalous coronary sinus and cardiac vein. 1. The greater (major) cardiac venous system (2) The smaller (minor) cardiac venous system (3)The compound cardiac venous system. The microanatomy of the various proper cardiac veins is not very well explained and illustrated in old or new literature; therefore, special attention is paid in the present study to the detailed microanatomy of the cardiac venous drainage. This includes the topograpy and structural and surface anatomy of the coronary sinus (position, length and shape, diameters, area of cross-section, circumference and volume, curvature, elevation, ostial angle, enlargement, duplication, absence), and the exact enternal and internal morphological landmarks of the coronary sinus with reference to its myocardial cover, isolated myocardial belts, and "free" myocardial cords which connect the atrial and ventricular myocardium, and the atrial ostium of the coronary sinus. It is established that the frequency, distribution pattern, courses and mode of opening of the major ventricular and atrial cardiac veins and the occurrence, morphology, and efficiency of the ostial valves of the coronary sinus and its tributaries all influence the success of any selective catheter implantation and venous reperfusion technique to a great degree. There are many peculiarities of the cardiac veins which are worthy of consideration, for instance intramyocardial and aberrant courses of the anterior interventricular vein, the oblique vein of the left atrium, the posterior interventricular vein, the small cardiac vein, the posterior vein of the left ventricle, the left and right marginal veins, and the anterior cardiac veins. Various forms and courses of the intramural venous tunnel, sinus or channel of the right atrium were found and illustrated, and discussed in terms of developmental and comparative anatomy. This review incorporates a great variety of clinically significant, new morphological findings with regard to the coronary sinus and the cardiac venous system. The many anatomical peculiarities and hindrances to the catheterization of the coronary sinus and the reperfusion of (even selected) cardiac veins are documented and evaluated; the various problems which may arise in venous reperfusion due to the presence of anatomical anomalies of the coronary sinus, cardiac veins, and ostial valves (of greater or lesser efficiency) are addressed. The presentation narrows a gap in the rather incomplete knowledge of the venous drainage of the human myocardium. PMID- 12645158 TI - The human nasolacrimal ducts. AB - The human nasolacrimal ducts are a borderland of the disciplines ophthalmology and otorhinolaryngology, which work close together in the treatment of nasolacrimal disorders. However, little knowledge exists concerning the physiology of the nasolacrimal system and, therefore, its pathophysiology. The aim of the present investigations was to obtain a better understanding of tear flow physiology, microbial defence strategies and pathophysiological processes by analysing the nasolacrimal system, thus opening new perspectives against existing therapeutical concepts of diseases of the nasolacrimal ducts. The human nasolacrimal ducts consist of the upper and the lower lacrimal canaliculus, the lacrimal sac and the nasolacrimal duct. They drain the tear fluid from the ocular surface into the lower meatus of the nose. The lining epithelium of the lacrimal sac and the nasolacrimal duct is faced by microvilli, and animal experiments in rabbits support the hypothesis that tear fluid components are absorbed in the human nasolacrimal system. Evidence of seromucous glands, intraepithelial lipids and a specific mucus layer leads to the conclusion that the normal three-layering of the tear film is also present in the nasolacrimal ducts. Based on its composition, the mucus layer, which is mainly synthesised by goblet cells and intraepithelial mucus glands, functionally serves as a simplified drainage of tear fluid, and it is attributed similar properties to epithelia of the gastrointestinal tract. Further defence mechanisms are represented by antimicrobial peptides IgA and immunocompetent cells (lymphocytes and macrophages) which show intraepithelially and subepithelially a special distribution. Moreover, organised lymphoid tissue is present, which reveals the cytomorphologic and immunophenotypic characteristics of mucosa-associated lymphoid tissue. The wall of the lacrimal sac and the nasolacrimal duct are made up of a helical system of different connective tissue fibres. Wide luminal vascular plexus are embedded in this helical system, which is comparable to a cavernous body. Caudally, the vascular system is connected to the cavernous body of the inferior turbinate. With distension the system may be "wrung out" due to its medial attachment and helically arranged fibrillar structures. Thereby, tear fluid is drained distally. The embedded blood vessels underlie vegetative control. By means of this innervation, the specialised blood vessels permit regulation of blood flow by opening and closing the lumen of the lacrimal passage as effected by the engorgement and subsidence of the cavernous body, at the same time regulating tear outflow. Related functions, such as a role in the occurrence of epiphora related to emotional responses, are relevant. Moreover, malfunctions in the cavernous body and in its innervation may lead to disturbances in the tear outflow cycle, ocular congestion or total occlusion of the lacrimal passage. Thus, the pathophysiology of primary acquired dacryostenosis can be explained: Descending inflammation from the eye or ascending inflammation from the nose initiates swelling of the mucous membrane, remodelling of the helical arrangement of connective tissue fibres, malfunctions in the subepithelial cavernous body with reactive hyperaemia, and temporary occlusion of the lacrimal passage. In the follow-up, repeated isolated occurrence of dacryocystitis leads to structural epithelial and sub-epithelial changes, which may lead either to a total fibrous closure of the lumen of the efferent tear duct or to a non-functional segment in the lacrimal passage that is manifest on syringing. The normally constant absorption of tear fluid components into the blood vessels of the surrounding cavernous body that are connected to the blood vessels of the outer eye could be a feedback signal for tear fluid production, which comes to a halt if these tear components are not absorbed. Thus, dry eye could be initiated. Defective stimulation of tear duct-associated lymphoid tissue (TALT) could result in abnormal immune deviation at the ocular surface leading to an autoimmunologic response that could cause dry eye pathology. PMID- 12645159 TI - Evaluation of neonatal tetanus elimination in Rajastan, India. PMID- 12645160 TI - GERD like symptoms as seen in a village. AB - Twenty cases of protozoal infection presented with symptoms suggesting GERD. Treatment with antacids and H2 blockers was unsuccessful in giving them relief. As they had also protozoal infection, treatment with anti-protozoal drugs gave them complete relief in the follow up period of one year after the end of treatment. Hence we have named these conditions simulating peptic ulcer GERD as pseodogerd or protozoal GERD. It is suggested that the person presenting with the symptoms of GERD in our area should be investigated for protozoal infection or should be given treatment of protozoal infection rather than that of peptic ulcers. It is also suggested that the length of treatment of protozoal infection should not be less than one month and not for three or 7 days as suggested in western text books. PMID- 12645161 TI - Correlation between chemical composition of biliary calculi and sera of stone formers. AB - A quantitative chemical analysis of total cholesterol, bilirubin, calcium, inorganic phosphate and iron of three types of biliary calculi (cholesterol, pigment and mixed) of 40 gall stone former was carried out and correlated to with those of there sera. A moderately positive correlation for inorganic phosphate, Ca2+ and Fe2+ content of sera and calculi of cholesterol stone patient was found. A Good positive correlation for total cholesterol, a moderately positive correlation for bilirubin and iron but no correlation for inorganic phosphate and calcium content of sera and calculi of pigment stone patient was observed. A good moderately positive correlation for iron but no correlation for total cholesterol, bilirubin, inorganic phosphate and Ca2+ content of sera and calculi of stone patient was found. PMID- 12645163 TI - An unusual case of biliary atresia. AB - Intrahepatic biliary cysts are rarely seen in the patients with biliary atresia. We describe a ten-month-old child with biliary atresia in whom the abdominal imaging studies (ultrasonography, computed tomographic scan and magnetic resonance cholangiopancreatography) revealed multiple intrahepatic biliary cysts ('bile lakes'). The child also had intrapulmonary shunting of blood due to pulmonary arteriovenous fistulae, which were demonstrated on contrast-enhanced echocardiography. Both these findings, 'bile lakes' and pulmonary arteriovenous fistulae occur rarely in biliary atresia. PMID- 12645164 TI - External fixator in the management of trochanteric fractures in high risk geriatric patients--a friend to the elderly. AB - The best treatment option for trochanteric fracture in a geriatric high risk patient with all associated medical and surgical problems remains debatable. Conservative methods of treatment are associated with dangerous complications of prolonged recumbency while open reduction and internal fixation under anaesthesia significantly increases the mortality and morbidity rates. We treated 110 elderly patients who were unfit or high risk cases for anaesthesia and major surgery for internal fixation due to associated medical and surgical conditions, by external fixation under local anaesthesia. The average age was 65 years and mean follow up was 18 months. 83.3% were ambulatory with support and 97.2% were able to manage activities of daily living at the time of discharge. At 18 months post surgery, 74% were ambulatory with a stick or better. The fracture united in an average of 16.4 weeks. Overall satisfaction rate was 80% at end follow up. The mortality rates were comparable to series of open reduction and internal fixation. Pin tract infection and knee stiffness were the major complications. External fixation done under local anaesthesia offers advantages in the form of a quick, simple relatively inexpensive procedure with negligible blood loss, preserves fracture haematoma, can be easily removed as an out patient procedure, besides it provides earliest possible ambulation and day care to the elderly high risk patient. PMID- 12645162 TI - Management of attention deficit/hyperactivity disorder--use of an effective paradigm. AB - The management of ADHD poses a great problem before the psychologists, psychiatrists and different behavioural scientists. The multi-model approach, combining pharmacologic with different non-pharmacologic interventions, is more effective than any form of therapy. In the present study the theta feedback, a non-pharmacologic technique, has shown beneficial role among the low medicated ADHD cases. The present study also signifies the role of multi-model intervention in the management of ADHD. PMID- 12645165 TI - Bacteraemia in high-risk patients. AB - In this study, we noticed a high incidence of bacteraemia in high-risk patients especially due to nonfermenter gram negative bacilli (NFGNB) and coagulase negative staphylococci (CoNS). Bacteraemia caused by some rare bacteria such as Moraxella spp., Aeromonas spp., Flavobacterium meningosepticum was also noted during the study. Antibiotic resistance pattern showed that many isolates were Multi Drug Resistant (MDR). This can be attributed to nosocomial-infection, which may occur due to more and more invasive procedures for diagnosis and therapy during long stay of patients in the hospital. Also, the MDR strains and the wide spread oxacillin resistant CoNS (OR-CoNS) and slowly emerging vancomycin resistant CoNS (VR-CoNS) associated with nosocomial infections pose a great threat to the clinicians. We recommend the usage of commonly used antibiotics along with cephalosporins for the patients admitted to these high-risk units, before the antibiotic susceptibility test findings. PMID- 12645166 TI - Comparative evaluation of efficacy and safety profile of three anti-tuberculous regimens in Mangalore. AB - The aim of our study was to evaluate and compare the therapeutic efficacy & safety profile of three different antituberculous regimens for pulmonary tuberculosis. The study sample size included 90 newly diagnosed, sputum positive patients of pulmonary. tuberculosis. 30 each from different groups. The parameters studied were, therapeutic efficacy included weight gain, cough, sputum examination and safety profile: nausea, vomiting, anorexia, gastritis, hepatitis, jaundice diarrhoea, rashes, dizziness, tingling & numbness, flu like symptoms & joint aches. Group-I showed statistically significant weight gain when compared to Group-II. Improvement in cough and conversion to smear negative were seen in 100% of patients in Group-I, 83.3% of patients in Group-II and 93.3% of patients in Group-III. Therapeutic efficacy was highest with Group I regimen, followed by Group III and Group II which was least efficacious. Group II also registered; the maximum cost and highest incidence of adverse effects. PMID- 12645167 TI - Colonisation of methicillin resistant Staphylococcus aureus among health care workers in a tertiary care hospital of Delhi. AB - Methicillin resistant strains of Staphylococcus aureus have been responsible for many nosocominl epidemics. Colonised employees of hospital such as asymptomatic nasal and hand carriers act as reservoirs for the spread of this organism within hospital. During a period of three months 150 HCW were screened for nasal and hand colonisation by MRSA. 6.6% (10) HCW were colonised by MRSA and 50% of these were carrying MRSA on both hands and anterior nares. 30% (3) of MRSA strains were multidrug resistant and 60% (6) were resistant to ciproflox. Whether the eradication of MRSA carrier state will lead to decreased rate of MRSA infection is yet to be documented. PMID- 12645169 TI - Methicillin resistance among isolates of Staphylococcus aureus: antibiotic sensitivity pattern & phage typing. AB - Out of 3988 clinical specimens from hospital admitted patients 230 strains of Staphylococcus aureus were isolated, 45 strains (19.56%) were Methicillin resistance Staphylococcus aureus (MRSA). All MRSA strains were beta lactamase producers. Multidrug resistance was observed among MRSA strains more commonly than in methicillin sensitive strains of Staphylococcus aureus (MSSA). Maximum strains were resistant to penicillin (100%), co-trimoxa zole (97%) & chloramphenicol (93.33%). As least resistant to gentamicin & ciprofloxacin shown by MRSA, these drugs can be used in few situations after susceptibility test. All strains of MRSA were sensitive to vancomicin (100%). Majority of strains (34 out of 45) showed MIC values of 4 ug/ml. Twenty eight out of 44 strains were non typable using routine phages. Study revealed that MRSA with associated multidrug resistance is common in this region. There is need to develop local set of MRSA phages for improvement of typability. PMID- 12645168 TI - Prevalence of fungal meningitis among HIV positive & negative subjects in Indore (MP state). AB - The aim of the study was to find out the prevalence of fungal meningitis among AIDS cases and to assess the prognosis of fungal meningitis among HIV positive & negative subjects. The study comprised of 15 & 10 cases of fungal meningitis among HIV positive & negative subjects respectively during the study period 1992 2001. India ink preparation and Gram's staining procedures were carried out on the centrifuged CSF deposits. The CSF deposits were also used for bacterial and fungal culture. In the present study the prevalence of fungal meningitis was noted among 15 (3.1%) of 483 AIDS cases. Twelve of them had cryptococcal meningitis while 3 were infected with Candida albicans. Four AIDS cases presented fungal meningitis as a primary opportunistic infection and HIV status was confirmed in 4 of them after the diagnosis of fungal meningitis. 13 of the 15 cases were in the age group 26-40 yrs while one was 55 yrs old and the other 16 month old child; these two cases had blood transfusion transmitted and vertically transmitted mode of HIV transmission respectively. Further, only two of 15 cases were females and both acquired HIV infection through blood transfusion. Overall prognosis of fungal meningitis was poor among HIV positives and 7 of the 15 cases died in hospital within 2-3 weeks after diagnosis of cryptococcal meningitis. Among HIV negative subjects, ten cases of fungal meningitis (3 with Candida albicans and 7 with Cryptococcous neoformans) were noted at our end and nine of them had immunocompromised status (3 cases of renal transplant on immunosuppressives, 3 cases neonates/infant and 2 diabetic subjects. The fungal meningitis is one of the important causes of morbidity & mortality among immunocompromised among HIV positive subjects. PMID- 12645171 TI - [Drug addiction: definition and classification]. AB - Basic aspects of drug abuse are discussed, from a semantic and physiopathologic point of view: drug definition; drug abuse and drug addiction; drug as a tool, that produces adverse or beneficial effects according to its use, rather than its intrinsic features. Simple and complicated drug-related problems are discussed in some depth, the former being associated with external factors, the latter with organic or mental disease. Practical implications of this concept are exposed. Similarities of drug abuse and suicide are underlined. It is concluded that ethical judgments and measures require an additional evaluation of these aspects, with particular reference to the medical component of drug abuse. PMID- 12645170 TI - ANCA: anti-neutrophil cytoplasmic antibodies and their role in vasculitis associated kidney disorders. PMID- 12645172 TI - [Bioethics and drugs: what is the significance of ethical aspects?]. AB - This philosophical point of view aims at offering a description of some ethical issues arisen from this question. They concern three values: personal freedom and responsibility, public security and benefits for all people, and a correct relationship between each other. An analysis is needed taking into consideration those different approaches, aimed at looking for best solutions: scientific, pragmatic, personal. This study gives moreover a brief account of the Italian bills concerning drug abuse new treatments. The reason for separating ethics issues from the rest of the debate also throws light on the possible synthesis of values, justice and law, as it is contemplated in the Italian Constitution. PMID- 12645173 TI - [Weakness of will]. AB - Addictive behaviours raise the issue how it is possible that a free agent is not able to act according to his own will. Indeed an agent can be defined as "addicted", to a substance or a behaviour, only when he does not succeed in discontinuing despite his will. Namely in the very idea of addiction it is involved the idea of weakness of will. The problem of weakness of will is one of the most intriguing problems posed by human spirit. This paper intends to show some of its manifestations. PMID- 12645174 TI - [Double diagnosis or comorbidity? Definition and clinical observation]. AB - There is a high prevalence of a comorbid substance use disorder in subjects with psychotic or mood disorders. Psychiatric patients are reported to abuse alcohol or drugs. This comorbidity raises the question whether one disorder is a consequence of the other. The self-medication hypothesis of substance abuse suggests that drug abuse is driven by an attempt to alleviate specific psychological distress but this unidirectional causality is not supported. The main aims of this review are to describe the difference between dual diagnosis and comorbidity in psychiatric patients and the possible patterns of different clinical conditions. PMID- 12645175 TI - [Psycho(patho)logy of "addiction." Interpretative hypothesis]. AB - This work deals with the possible role of psychopathological features as for the pathogenesis of substance use disorders, across three distinct stages of its spontaneous course: initiation, transition to habit, that is becoming accustomed to regular substance use, and eventual self-maintaining addiction. Reviewed data from literature are discussed concerning personality characteristics of addicted patients and the issue of addictive personality. On such basis, we propose a classification of addictive styles according to our personal experience in the field (environmental reaction, self-medication, and metabolic automacy) in the attempt to better illustrate the dynamics of the various stages of the abuse, each accounted for as a distinct milestone along the same path of pathogenesis. In fact, interpretative models elaborated so far, though partly useful in providing with a plausible explanation for special subpopulation of addicts, failed to portray an overall shared dynamic. In other words, no satisfactory explanation has been so far provided to justify the basic development of the addictive condition along the bare events observed along its course. Our integrated model especially accounts for the issue of mood disorders as the a weightsome psychic substrate for the development of a substance-resorting attitude. PMID- 12645176 TI - [Family communication and primary prevention]. AB - A good communication within the family is the basis for the primary prevention of discomfort and deviance. With the aim of prevention of the frequent acts of violence within the family, the present article responds to the request of behavioural guidelines in schools, families, communities, and other pedagogical settings. Thus, the delicate issue of self-esteem in adolescents, the dysfunctional parental behaviour and the symptoms of discomfort of the teenagers are discussed. Besides the behavioural strategies for parents, the hot topic of alcohol abuse, which is cause of a significant number of car accidents, is briefly mentioned. PMID- 12645178 TI - [MDMA (ecstasy): neuroendocrine and behavioral features]. AB - Long lasting 5HT system impairment has been demonstrated in experimental animals exposed to ecstasy use; MDMA seems to be able to induce behavioral conditioning and reiterated use because of its dopaminergic action. Among behavioral aspects of ecstasy users mood disorders, irritability and difficult in relationships, interpersonal difficulties, high levels of impulsiveness and hostility, high sensation seeking, cognitive and attentive deficit have been reported. A derangement of serotonin system function was reported also in humans exposed to ecstasy, as confirmed by neuroendocrine challenges and brain imaging techniques. Recent researches suggest functional changes in dopaminergic system too. The persistence of behavioral and neuroendocrine changes many months after MDMA's discontinuation, indicate a lack of reversibility in the dysfunction induced by ecstasy, or the persistence of psychobiological traits that could preexist to MDMA exposure, possibly involved in substance abuse vulnerability. PMID- 12645177 TI - [Drugs and pregnancy]. AB - The evidence of the transplacental transfer of most of the drugs administered to the mother has changed the old concept of the invulnerability of the fetus due to the placental barrier protection. The commonly used drugs act on the fetus by means of two different mechanisms: directly after the placental transfer, and indirectly influencing the utero-placental circulation and maternal homeostasis. Fetal effects can be several and they depend on the type of the drug, dosage, route of administration, gestational age in relation to the fetal ontogenesis, and on the amount of drug which reaches the fetus. For example, some drugs are redistributed in the maternal compartment; others remain in the fetal compartment by virtue of their chemical-physical characteristics and/or altered fetal homeostasis, i.e., fetal acidosis results in ionisation of the loco-regional anesthetics, which cannot leave the fetus, and therefore are entrapped in the fetal compartment. The complexity of the fetal-maternal pharmacology has implemented research on the animals, and epidemiologic and clinic studies. In spite of the research performed in the last years, the long-term perinatal effects are still to be clarified for many drugs administered to the mother. PMID- 12645179 TI - [Psychobiological determinants of risk behavior in adolescence]. AB - Human adolescence is mainly characterised by an elevated number of psychological, biological and behavioural changes. This rapid as well as challenging change in the "inner state" during adolescence in humans seems to share a number of similarities with the age-dependent alteration of the normal and "linear" development in other mammalian species. Hence, in order to further investigate the biological bases of these idiosyncratic age-related responses, it seems worth to develop and to study suitable animal models for this developmental phase. Aim of the present review is to show experimental results indicating an elevated similarity between humans and rodents (mice and rats) in a number of psychological (elevated levels of novelty seeking as well as of harmavoidance due to a reduced anxiety) behavioural (elevated exploration of novel as well as potentially dangerous environments and peculiar vulnerability to incentive properties of drugs of abuse) and neurobiological (developing state of the underlying cortical pathways implicated in the above mentioned responses) processes. These findings could prove really useful in view of a general deeper comprehension of this age period and helpful in developing earlier and more effective prevention strategies aimed at reducing the social impact of dangerous and reckless behaviours frequently shown by human adolescents (drug addiction, undesired pregnancy. AIDS and car accident). PMID- 12645180 TI - [Effects of drug abuse on sexual response]. AB - Drugs of abuse, like alcohol, opiates, cocaine and cannabis, are used by many young people for their presumed aphrodisiac properties. It is well known, instead, that, apart from the subjective effects, they negatively affect the sexual response. Alcohol has direct toxic effects on the gonads (testes and ovaries) and the liver (it increases the catabolism of testosterone and its transformation in estrogens). Besides, it inhibits the hypothalamus-pituitary gonads axis (HPG). The opioids inhibit the HPG axis and increase the prolactin levels which, in turn, interferes with the male and female sexual response. The acute effects of cocaine are stimulants mainly for its dopaminergic properties but in the long run it causes sexual dysfunctions (erectile, etc.) mainly due to hyperprolactinemia. Cannabis, at high doses, could inhibit the HPG axis and reduce fertility. The knowledge of these effects should be better disseminated among subjects at risk for deterrent purposes. PMID- 12645181 TI - [Infections and other causes of mortality correlated with drug addiction]. AB - Drug use and related injection practices determine an excess mortality in drug abusers compared to the general population adjusted for age and gender. HIV/AIDS and overdose are the most important causes of death among injecting drug users; however, excess mortality is also due to other causes, such as cirrhosis and endocarditis. Infections have a great impact on the health of injection drug users. Viral infections, such as HIV, HCV, and HBV, are the most important ones, but bacterial and fungal infections, and, to a lesser extent, protozoal infections, are also common. In depth knowledge of drug-related morbidity and mortality causes is important to improve clinical skills and to implement control activities. PMID- 12645182 TI - [Procedural aspects and interpretation problems in the analysis of drugs of abuse]. AB - Addiction scene is showing rapid changes about substances and patterns of use/abuse. Increasing requirements from clinical and forensic fields to identify drugs of abuse and to find them at ever decreasing levels in biological samples, have led to a great improvement in the state of the art of analytical toxicology. This paper discusses various aspects of these developments highlighting possibilities and problems that make the toxicological determination to be evaluated with prudence, keeping in mind that these results involve legal aspects too. Special attention is focused to the so-called "designer drugs", to the outside laboratory assays and to the problems due to poly-drugs assumption. PMID- 12645183 TI - [Illicit market of controlled drugs in Italy: new drugs and trends]. AB - The data related to abuse and seizures of the main classes of controlled drugs in Europe and Italy are presented. The system of keeping up-to-date the tables of controlled drugs in Italy is briefly discussed. In the end, a list of the main new drugs in the clandestine market of synthetic drugs in Italy is presented, by taking into account the street names, doses and pharmacological effects. The paper ends with a brief description of the new trend of purchasing controlled drugs on internet sites. PMID- 12645184 TI - [Recent initiatives of the European Union on the field of drug use and trafficking]. AB - Moving from the general framework offered by the Treaties of the European Union and European Community, the paper presents the more recent specific initiatives of the Union in the field of fight against drugs and in particular on the European Union Plan of Action, adopted by the European Council in June 2000, and on the need for a global and balanced approach which it proposes as the main focus of the action of the European institutions. The paper then examines separately the initiatives taken both at the level of the reduction of the demand, mainly at the preventive and sanitary level, and of the supply reduction, of a mainly repressive character. Conclusively, the main initiatives in the field of international co-operation among the European Union and third Countries. PMID- 12645185 TI - [Drug trafficking and international law]. AB - The production, the commerce and the use of drugs and other substances are ruled by several conventions of international law, that, at first, have had as object the production and the commerce of drugs for lawful purposes, and the measures required to prevent and to repress, at certain conditions, abuses and unlawful traffics. Just more recently, following some solicitations noticed by the International Community, and according to the concept of "well-balanced approach" described in the text, the measures introduced in this way were supported by a more incisive international movement, fit for the repression of unlawful traffics of drugs and to the adoption of suitable measures of prevention, also to avoid, at national level, sanitary, social and economical implications of the criminal phenomenon. PMID- 12645186 TI - Identification of di(beta-phenylisopropyl)amine as the main ingredient in illicit amphetamine tablets. AB - The identification of di(beta-phenylisopropyl)amine found as the main ingredient in several sets of amphetamine tablets sold on the illicit market in Rome, Italy, during 1999-2000 is described. The identification was achieved by examining the ultraviolet and nuclear magnetic resonance spectral properties as well as the chromatographic, gas chromatographic, and mass spectrometric data. The molecular structure of the ingredient showed a close analogy to the amphetamine and could very likely produce similar pharmacological activity. A preliminary test on the metabolic pathway of di(beta-phenylisopropyl)amine performed on rats, suggests its biotransformation to amphetamine. PMID- 12645187 TI - Respiratory response to carbon dioxide stimulation during low flow supplemental oxygen therapy in chronic obstructive pulmonary disease. AB - BACKGROUND AND PURPOSE: Oxygen supplementation is the treatment most commonly used to relieve dyspnea in chronic obstructive pulmonary disease (COPD). There is a lack of data, however, on the response of the respiratory drive to low flow oxygen in severe stable COPD. The purpose of this investigation was to evaluate the magnitude of chemoresponsiveness to low flow supplemental oxygen in patients with COPD of variable severity in terms of mouth occlusion pressure at 100 msec (P0.1), P0.1 and minute ventilation (MV) response to CO2 stimulation, and blood gas tension. METHODS: Twenty-six patients with stable COPD of variable severity were divided into two groups: those with mild airway obstruction and normocapnia (n = 14) and those with hyperinflation and hypercapnia (n = 12). RESULTS: Arterial oxygen tension, oxygen saturation, and arterial CO2 tension were significantly increased after oxygen therapy in COPD patients with or without hypercapnia (all p < 0.01). COPD patients with hypercapnia had a significantly higher P0.1 (0.7 +/- 0.07 kPa) than those with normocapnia (0.3 +/- 0.03 kPa, p < 0.01). Oxygen significantly decreased the P0.1 adjusted by end tidal CO2 pressure (delta P0.1/PETCO2) only in patients with hyperinflation and hypercapnia, from 0.2 +/- 0.05 to 0.1 +/- 0.03 kPa (p < 0.05). There was a weak correlation between P0.1/PETCO2 and forced vital capacity (FVC; r = 0.41, p < 0.05) or forced expiratory volume in 1 second (FEV1; r = 0.45, p < 0.05). In addition, the arterial CO2 tension (PaCO2) was inversely related to P0.1/PETCO2 (r = -0.57, p < 0.01). The MV with 6% CO2 (MVCO2) was also significantly decreased in the hypercapnic group from 17.9 +/- 3.7 to 14.8 +/- 4.9 L after oxygen therapy (p < 0.01). The maximum inspiratory pressure did not change after oxygen usage in either group. CONCLUSION: We conclude that short-term oxygen therapy may blunt respiratory response to CO2 in COPD with chronic hypercapnia. Cautious observation of respiratory response is needed during oxygen therapy in COPD patients with a higher magnitude of air-trapping and hypercapnia. PMID- 12645188 TI - Efficacy of low-dose mycophenolate mofetil therapy for Taiwanese renal transplantation patients receiving primary cyclosporine immunosuppression. AB - BACKGROUND AND PURPOSE: Mycophenolate mofetil (MMF) in combination with cyclosporine or tacrolimus prevents acute rejection and chronic allograft failure in renal transplantation in Western countries. We began to add low-dose MMF to primary cyclosporine immunosuppressive therapy in renal transplantation at the Department of Surgery of National Taiwan University Hospital in 1998. This study compared low-dose MMF to conventional therapy in Taiwanese renal transplant recipients. METHODS: This retrospective cohort study determined the efficacy of low-dose MMF therapy (1 g/day in divided doses). A total of 275 cases with allograft kidney transplants were grouped according to whether they received transplants before or after the adoption of MMF therapy (Period I: 1987-1993; Period II: 1994-1997; Period III: 1998-September 2000). The prognostic significance of MMF therapy and graft and patient survival rate in each time period were assessed. RESULTS: The 18-month graft survival rate was 84.9% in Period I, 86.3% in Period II, and 91.9% in Period III. The 5-year graft survival rates in Periods I and II were 69.3% and 76.6%, respectively. Acute rejection was significantly detrimental to graft survival (p = 0.048), while MMF therapy was significantly advantageous to graft survival (p = 0.015); treatment when MMF was available was also significantly associated with better graft survival (p = 0.043). There was a negative correlation between acute rejection and graft survival (p = 0.035); MMF therapy produced a protective effect on graft survival independent of acute rejection (p = 0.010). CONCLUSION: Low-dose MMF therapy significantly improved graft survival after renal transplantation in Taiwanese kidney allograft recipients. PMID- 12645190 TI - Enzyme replacement therapy with imiglucerase in Taiwanese patients with type I Gaucher disease. AB - BACKGROUND AND PURPOSE: The management of Gaucher disease has been dramatically improved by the development of enzyme replacement therapy (ERT), but there have been no reports of imiglucerase treatment of this disorder in Taiwanese patients. The goal of this study was to evaluate the outcome of imiglucerase treatment for type I Gaucher disease in the first Taiwanese patients to receive this treatment. METHODS: Six patients received ERT for type I Gaucher disease at National Taiwan University Hospital. Their mean age was 36.5 years; four had undergone splenectomy before and one after the start of ERT. All received intravenous imiglucerase 60 U/kg every 2 weeks. Patients were regularly evaluated, including physical examination, laboratory tests and bone roentgenography. RESULTS: All six patients responded well to therapy. The mean length of therapy was 30.3 months. Five patients had anemia, and the mean increase in hemoglobin was 4.46 g/dL. Five patients had thrombocytopenia, and the mean increase in platelet count was 259 x 10(3)/microL. Laboratory data showed a slower response in hemoglobin than in platelet count, which may have been related to splenectomy and low initial platelet count. A decrease in serum alkaline phosphatase was the most prominent finding related to liver function. All patients had a prominent decrease in liver size. No patient had painful bony crises, but no significant improvement was observed in skeletal roentgenograms. There were no serious side effects. CONCLUSION: ERT reversed the visceromegaly and hematologic abnormalities of Gaucher disease and improved the patient's quality of life. It was effective in reducing organomegaly, correcting anemia and thrombocytopenia, improving liver function and alleviating bone pain. PMID- 12645189 TI - Latex allergy and latex-fruit syndrome among medical workers in Taiwan. AB - BACKGROUND AND PURPOSE: Latex allergy is a serious occupational health problem among medical workers, but there are few data regarding its incidence and clinical characteristics in Taiwan. No data on the latex-fruit syndrome in Taiwan have been reported. This study investigated the prevalence of both latex allergy and latex-fruit syndrome, and risk factors for the development of latex allergy among medical workers. METHODS: A total of 302 medical workers who had daily contact with latex gloves during work at three hospitals in central Taiwan were interviewed and screened by questionnaire to detect allergic reactions to latex. Those with a history of immediate allergic reaction to latex gloves were screened for specific IgE antibodies against latex. Subjects whose screening results were positive were considered to have immediate latex allergy. Latex-fruit syndrome was defined as a clinical condition that manifested as immediate allergic reactions to both latex and fruit. RESULTS: Twenty-six (8.6%) medical workers had immediate allergic reactions to latex. The severity of latex allergy was correlated to the serum level of latex-specific IgE. Seven subjects with latex allergy (26.9%) had latex-fruit syndrome. Risk factors for development of latex allergy among medical workers were latex glove-related hand dermatitis, fruit allergy, preexisting atopic disease, hospital employment for more than 2 years, and total exposure to latex gloves of more than 9,000 hours. CONCLUSION: Latex allergy is an underrecognized occupational health problem in Taiwan. Early identification of medical workers at high risk is important so that they can be advised to use non-latex gloves as early as possible to prevent further sensitization to latex proteins. PMID- 12645191 TI - Cost-effectiveness analysis of interferon-alpha therapy in the treatment of chronic hepatitis B in Taiwan. AB - BACKGROUND AND PURPOSE: Although interferon-alpha has been approved for chronic hepatitis B treatment in Taiwan, the high price necessitates study of its cost effectiveness to define its role from a burden of disease perspective. METHOD: A stimulation model was constructed to project the long-term effects of interferon treatment or standard care for a hypothetical group of 35-year-old chronic hepatitis B patients in Taiwan. Data on interferon treatment efficacy, disease progression, vital statistics, medical expenditure for various liver disease states, and quality-of-life measures based on Taiwan data were collected and used in the model. RESULTS: The model indicated that a 35-year-old chronic hepatitis B patient treated with interferon in Taiwan would have a life expectancy of 29.08 years, versus 28.67 years for the same patient who did not receive interferon. Corresponding to this gain in life expectancy of 0.41 years (or 0.18 quality adjusted life-years, QALYs), the incremental cost-effectiveness ratio (ICER) was 492.000 New Taiwan dollars (NT$; 14,200 US$ at 34.7 NT$/US$) per QALY with an annual 3% discount rate. Monte Carlo simulation taking into account the range of plausible values for all model parameters indicated that 95% of the ICERs were in the range of 65,000 NT$ (1.900 US$)/QALY to 683,000 NT$ (19,700 US$)/QALY, and 90% of the ICERs were below 413,000 NT$ (11,900 US$)/QALY. CONCLUSION: Interferon has a favorable cost-effectiveness profile in the treatment of chronic hepatitis B in Taiwan. However, the study results cannot be interpreted in isolation and need to be compared with the cost-effectiveness profiles of treatment regimens for other major chronic diseases in Taiwan. PMID- 12645192 TI - Syphilitic uveitis as the initial manifestation of HIV infection. AB - Syphilis is an uncommon cause of uveitis in HIV-infected patients. We report a case of bilateral panuveitis and describe its characteristics as the initial manifestation of HIV infection. A 74-year-old heterosexual male complained of blurred vision and floaters in both eyes for 40 days. Slit lamp examination showed diffuse keratic precipitates and cells in the anterior chamber of both eyes. Fundus examination revealed multiple small white dots and scattered retinal hemorrhage over the mid-equatorial retina with marked vitritis. Physical examination disclosed multiple erythematous papules over bilateral palms compatible with secondary syphilis. Serologic tests--the venereal disease research laboratory (VDRL) test, fluorescent treponemal antibody absorption (FTA ABS) test, and Treponema pallidum hemagglutination (TPHA) test--were all positive. Aqueous fluid also showed positive FTA-ABS reaction. Under the impression of acquired secondary syphilis, enzyme-linked immunosorbent assay and Western blot test were performed and revealed concurrent HIV infection. After intravenous administration of penicillin-G, 18 million units daily for 2 weeks, the vitritis and retinochoroiditis improved. All patients with panuveitis of unknown cause should undergo VDRL and FTA-ABS screening. Subsequent testing for HIV antibody in leutic uveitis is also mandatory. PMID- 12645193 TI - Treatment of Toxoplasma brain abscess with clindamycin and sulfadiazine in an AIDS patient with concurrent atypical Pneumocystis carinii pneumonia. AB - Toxoplasmosis is the most common opportunistic infection of the central nervous system in patients with AIDS. The standard treatment for toxoplasmic encephalitis is pyrimethamine and sulfadiazine. There have been few reports of concurrent Toxoplasma brain abscess and cavitary Pneumocystis carinii pneumonia (PCP) in Taiwan. We report the case of a 26-year-old homosexual man with coexisting infection with Toxoplasma gondii and P. carinii who was successfully treated for brain abscess with clindamycin and sulfadiazine. The cavitary lung lesions, initially diagnosed as pulmonary tuberculosis, were proved to be PCP by lung biopsy. HIV infection and syphilis had been diagnosed 1 year before admission. He presented with general weakness, ataxia, nausea, blurred vision and fever for 2 weeks. Magnetic resonance imaging of the brain revealed multiple ring-enhanced lesions over the cerebrum and cerebellum. Chest roentgenography showed a 3-cm lesion with cavitation over the right upper lung field. Diagnostic computerized tomography-guided lung biopsy revealed P. carinii cysts. Clindamycin, sulfadiazine and trimethoprim (TMP)-sulfamethoxazole (20 mg/kg/day TMP) were given with good response. His CD4 count rose from 40 to 280/microL 4 months later. All antibiotics were discontinued after 4.5 months due to the development of a skin rash. He was well at follow-up 1 year later. This case suggests that the combination of clindamycin and sulfadiazine is an effective treatment for Toxoplasma brain abscess and highlights the importance of diagnostic lung biopsy for cavitary lung lesions, particularly in a region endemic for tuberculosis. PMID- 12645194 TI - Rigid spine syndrome with chronic respiratory failure. AB - Rigid spine syndrome (RSS) is a rare myopathic ailment characterized by mild axial and proximal muscle weakness. Muscle contraction in these patients causes limitation of neck and trunk flexion, scoliosis, and mild joint deformity. We report the case of a 30-year-old man with RSS who presented with severe restrictive ventilatory defect (forced vital capacity, 1.53 L, 39% of predicted), mild scoliosis (Cobb's angle 12), proximal muscle weakness and stiff back. Creatine phosphokinase was 986 IU/L. Muscle biopsy of the right vastus lateralis revealed increased variation in muscle fiber diameter, proliferation of endomysium, and type I fiber atrophy. Round and whorled fibers indicating myopathic change were found on sections stained with nicotinamide adenine dinucleotide dehydrogenase and succinate dehydrogenase. RSS was diagnosed based on myopathic findings and clinical presentation. Nocturnal chronic respiratory failure and fragmented sleep developed. He was treated with continuous positive airway pressure and had recovered his normal daily activity by 1-year follow-up. We suggest that patients with RSS should be assessed for possible ventilatory failure; treatment with nasal continuous positive airway pressure or bilevel continuous positive airway pressure therapy should be considered. PMID- 12645195 TI - Nail-patella syndrome with renal involvement and antecubital pterygia. AB - Nail-patella syndrome (NPS) is a rare, autosomal-dominant hereditary disorder characterized by nail dysplasia and multiple osseous abnormalities. Some patients may develop renal function impairment and even end-stage renal disease. We treated a 42-year-old female patient with proteinuria who presented with a web like structure over the antecubital fossa and hypoplastic patellae. In addition, she had other characteristic findings, including bilateral iliac horn, triangular nail lunulae and hypoplastic radial head. She had impaired renal function, and renal biopsy showed mesangial proliferative glomerulonephritis. Additional cases were found in her family. Her mother had most of the signs of NPS as well as advanced renal failure. Her elder sister had knee abnormalities without the web like elbow condition. Both daughters also had the characteristic features of NPS. During follow-up 30 months after the initial examination, the patient had stable renal function and mild proteinuria. PMID- 12645196 TI - Identification of Escherichia coli O157:H7 by multiplex PCR with primers specific to the hlyA, eaeA, stx1, stx2, fliC and rfb genes. AB - Escherichia coli O157:H7 can cause fatal diseases such as hemolytic-uremic syndrome and thrombotic thrombocytopenic purpura. The early symptoms of E. coli O157:H7 infection are similar to those of infection with other gastrointestinal bacteria, so the availability of a rapid and precise method to identify E. coli O157:H7 is important for early recognition. We report a multiplex polymerase chain reaction (PCR) with primers to detect the presence of the E. coli O157:H7 genes hlyA, which produces enterohemolysin, eaeA (intimin), stx1 (Shiga-like toxin I), stx2 (Shiga-like toxin II), fliC (flagella H-antigen), and rfb (surface O-antigen). The technique improves specificity compared to general PCR when used with > or = 10(3) CFU/assay. Specificity was examined using 32 E. coli O157:H7 strains from the USA, Japan, Canada and Taiwan, all of which were positive for hlyA, eaeA, fliC, rfb, and stx1 and/or stx2. Thirty-five non-O157 enterovirulent E. coli strains, two Shigella sp strains, and two Salmonella sp strains were also examined and none gave the positive gene expression results described above. This multiplex PCR was highly specific in identifying E. coli O157 and was more useful than general PCR in early clinical detection of this pathogen. PMID- 12645198 TI - Application of a laser flare-cell meter to examine blood-aqueous barrier function in a patient with retinitis punctata albescens. AB - A laser flare-cell meter was used to examine blood-aqueous barrier function in a 21-year-old male patient who had typical retinitis punctata albescens with white dots scattered throughout large areas of the fundus in both eyes. Fluorescein angiography showed multiple areas of hyperfluorescence over dot lesions and window defects. Electroretinogram demonstrated subnormal amplitude and electrooculogram showed a reduced light-peak/dark-trough ratio. Goldmann perimetry revealed constricted peripheral fields. Laser photometry showed an increased aqueous flare intensity in both eyes compared to results in 10 normal subjects. Quantitative measurement of the aqueous flare intensity by a laser flare-cell meter might indicate abnormalities in the blood-aqueous barrier in patients with retinitis punctata albescens. PMID- 12645197 TI - Ceftriaxone-resistant Salmonella enterica serovar Hadar: evidence for interspecies transfer of blaCMY-2 in a Taiwanese university hospital. AB - The emergence of resistance to extended-spectrum cephalosporins in salmonellae is an increasing clinical problem. We report the characteristics of a ceftriaxone resistant Salmonella enterica serovar Hadar strain collected in 2001 from a patient with a postoperative wound infection in a university hospital in Taiwan. Resistance to extended-spectrum cephalosporins was found to be due to production of the plasmid-mediated CMY-2 AmpC beta-lactamase. To our knowledge, this is the first report of S. hadar harboring blaCMY-2. Seven CMY-2-producing Escherichia coli isolates collected in 2000 were investigated for comparison. Conjugation experiments and plasmid analysis showed an identical plasmid carrying blaCMY-2 in both the Salmonella isolate and one E. coli isolate, suggesting the possibility that the Salmonella isolate acquired the resistance plasmid from E. coli. These findings suggest that measures are necessary to restrict antibiotic use and so prevent the spread and development of antibiotic resistance in Taiwan. PMID- 12645199 TI - [Complex treatment of varicosity of the lower extremities]. AB - Over 10 years 1296 patients with an uncomplicated form of varicose disease were treated with compressive sclerotherapy before surgery. Sclerotherapy was performed as the first stage of complex treatment in combination with various surgeries. Over the last 5 years Doppler ultrasound study and coloured angioscanning were carried out in each patient before treatment. There were a total 1457 surgeries. The majority of the patients (916) were examined 4 months to 6 years postoperatively. The results of the treatment were good in 91.6% patients, the rate of recurrence after surgery did exceed not 4.03%. This complex therapy enhances the cosmetic effect and shortens the time treatment without loss for radicality. PMID- 12645200 TI - [Prevention of purulent complications after reconstructive vascular operations in elderly and aged patients]. AB - Results of surgical treatment of 382 patients with III-IV stages critical limb ischemia and concomitant diseases were analyzed. The mean age of the patients was 67.7 years. All the patients underwent reconstructive operations on lower limbarteries. Three variants of antibiotic therapy were compared: 1) antibiotics were administered parenterally; 2) they were injected endolymphatically; and 3) they were used parenterally and endolymphatically. Combined antibiotic, i.e. a combination of parenteral and endolymphatic routes of administration was most effective. PMID- 12645201 TI - [One-stage correction of heart defect and funnel chest in the Marfan syndrome]. AB - The first experience of simultaneous operations for heart defect and funnel chest in adult patients with the Marfan syndrome is presented. One-stage correction permits simultaneous removal the underling (valvular disease) and concomitant (laterocaudal ectopia of the heart and central vessels due to deformity of the anterior thoracic wall) causes of hemodynamic disorders. It is concluded that this policy is expedient in strictly selecting patients. PMID- 12645202 TI - [Doppler study in the evaluation of results of varicocele treatment in teenagers]. AB - Results of treatment in 62 patients with varicocele were evaluated with anamnestic, physical and Doplerographic data. Recurrence of varicocele was seen after the Palomo surgery in 4.9% patients. Hydrocele reoccured after surgery in 2 teenagers. Doppler study revealed that 37.5% patients had orthostatic extravasal obstruction of the left renal vein in the aortomesenterial "forceps" (0.15 +/- 0.03 cm, p > 0.05) with a significant decrease in volumetric blood flow velocity by 44 +/- 30.01 ml/min. Postsurgical signs of venous renal hypertension were seen in 2 patients. Doppler ultrasound study more objectively evaluate long-term results of varicocele treatment. PMID- 12645204 TI - [Non-epithelial tumors of gastrointestinal tract: diagnosis and surgical treatment]. AB - Non-epithelial tumors of the gastrointestinal tract (GIT) are rare diseases, their rate is less than 2.6% of all tumors of this localization. Analysis of 271 clinical cases of non-epithelial tumors demonstrated that most of them (n = 174) belonged to smooth muscle tumors. Features of growth and localization of benign and malignant non-epithelial tumors were demonstrated. It was revealed that only morphologic study (frequently with histochemical methods) verified the diagnosis of neoplasm. The choice of the scope of surgical intervention depends on the localization, spread and morphology of tumor. Difficulties of verification before surgery lead to active surgical policy for non-epithelial GIT tumors. PMID- 12645203 TI - [Correction of urination disorders caused by benign prostatic hyperplasia in cardiac surgery]. AB - Results of treatment in 14 patients with obstructive benign hyperplasia (adenoma) hospitalized for aortocoronary bypass surgery and coronaroplasty were analyzed. The baseline volume of residual urine ranged from 60 to 250 ml; uroflowmetry, from 4 to 8 ml/sec. During 7 day, the patient were treated with Omnic 0.4 mg a day, daily massage of prostate. Treatment substantially decrease residual urine volume and increased uroflowmetry indices. In 5 cases, the antihypertensive effect of Omnic was used for the treatment of arterial hypertension frequently accompanied by ischemic heart disease. This treatment improved urination and prevented acute ischuria before and after surgery. PMID- 12645205 TI - [Biophysical properties of bile and urine in patients with cholelithiasis]. AB - The electric conductivity of the hepatic bile received during and 24 hours after surgery from the drainage introduced into the common hepatic duct was studied in 94 patients with cholelithiasis. It is demonstrated that conductivity increases in cholestasis, inflammatory process in the biliary tract and depends on the hepatic morphofunctional status. The specific electric conductivity of intraoperative bile that is more than 192 S/m is considered to be criteria for the diagnosis of cholangitis. The electric conductivity of urine before and in first 4 days after surgery decreases in inflammation of the biliary tract and depends on the degree of concomitant renal dysfunction and electrolyte disturbances. There was a decrease in the conductivity of bile and urine in all studied groups. Detection of biophysical parameters of biological fluids may objectively control the postoperative period and correct treatment policy. PMID- 12645206 TI - [Rationale for removal of fibrous capsule in echinococcectomy from the liver]. AB - Experience of radical surgical treatment in 253 patients with hepatic echinococcosis is presented. Twenty-six patients underwent hepatic resection; 218, pericystectomy, 9 patients, combined interventions. Based on comparative analysis of the results of surgical treatment in 109 patients from the study and control groups, it is concluded that radical operations have significant advantages and show fever postoperative complications and recurrences. Removal of fibrous capsule is a token of successful treatment for hepatic echinococcosis. PMID- 12645207 TI - [Comparative aspects of surgical treatment of hemorrhoid]. AB - Based on experience in treating 64 patients with chronic hemorrhoid a comparative clinical evaluation of efficiency of electrosurgical methods is carried out hemorrhoidectomy. Compared with traditional Milligan-Morgan surgery, electrosurgical method demonstrates the absence of intraoperative blood loss, a reduction in the algetic syndrome after surgery, a decrease in postoperative complications rates by 4.8 times. Physiological and physical activity, stool are also recovered by electrosurgery. PMID- 12645208 TI - [Results of ligature of distal branches of the upper rectal artery in chronic hemorrhoid with the assistance of Doppler ultrasonography]. AB - In October 2000 to February 2002 Doppler ultrasonography assisted suturing and ligation of hemorrhoidal arteries were performed in 102 patients with chronic hemorrhoids (57 men, 45 women, mean age 44 +/- 4.3 years). The mean duration of the disease was 10 +/- 4.6 years. Elimination of scarlet blood and hemorrhoidal prolapse without concomitant proctologic diseases were indications for surgery. Sixty-nine patients were followed up for more than 12 months, the follow-up lasted on the average 12 +/- 4 months. There were no clinical symptoms in 57 (82.6%) on the 69 patients. Combined ligation of arterial branches and nodes with latex rings was performed in 7 patients with stages III-IV hemorrhoids. Sclerotherapy was performed in 2 patients with stage III with occasional elimination of blood on defecation. Hemorrhoidectomy was performed in 3 (2.9%) patients with stage IV hemorrhoid due to ineffective ligation. Suturing and ligation of distal branches of the upper rectal artery decrease the inflow of arterial blood to hemorrhoidal nodes without venous outflow disorders. Fixation of hemorrhoidal nodes to the muscular wall eliminates the symptoms of hemorrhoidal prolapse. These factors lead to elimination of hemorrhoidal symptoms in 82.6% of the patients operated on. PMID- 12645209 TI - [Preoperative care and surgical treatment of small-intestinal fistulas in children]. AB - Experience in using complex treatment in 42 children with unformed small intestinal fistulas, generalized defects of the anterior abdominal wall and severe disorders of alimentary status is presented. Their preoperative preparation included parenteral alimentation, enteral probe feeding with special elementary milk formulas, correction of metabolic disorders. Intestinoplication during surgery was carried out with the medicinal glue MK-7 to prevent intestinal obstruction and formation of interloop abscesses. The use of portions of the tendinous part of the musculus tensor fasciale late was one the simple and effective methods for plasty of anterior abdominal wall defects. PMID- 12645210 TI - [Physical methods of dissection and coagulation in surgery]. AB - The paper presents 20-year experience in using various devices for tissue dissection and coagulation of such as electrosurgical ones EN-57M, Efa-0201 (Russia), Karl Storz-endoscope (Germany), Force-300, Force-FX, LigaSure ("Valley lab", USA), Erbotom ICC 350 ("Erbe"), Argo-3000 ("Soring", Germany), a Surgitrone radioknife ("Ellman", USA), ultrasonic Auto Sonix devices ("Auto Suture", USA), a harmonic Ultracision scalpel ("Ethicon Endo-Surgery", USA), laser Scalpel-1 and Romashka-1 devices (Russia), plasma Fakel-01 and Prometey devices (Russia). The results of 424 experimental studies and clinical application of different energies in 1683 patients are analyzed. The influence of different types of energy on tissue and reparative processes were revealed, morphological changes were studied. Advantages and shortcomings of each type of energy are demonstrated, recommendations for their optimum application are formulated. PMID- 12645211 TI - [Current methods for local drug therapy of infected wounds]. PMID- 12645212 TI - [Posttraumatic aortocaval anastomosis]. PMID- 12645214 TI - [A rare complication of acute pancreatitis]. PMID- 12645213 TI - [Combination of synchronic-metachronic multiple primary cancer of cardia, colon with maxillofacial and oral neoplasia]. PMID- 12645215 TI - [Aspects of abdominoplasty]. PMID- 12645216 TI - [In the memory of Prof. N.I.Napalkov]. PMID- 12645218 TI - [Clinical impact of ultrasonography between the 10th and the 13th week of pregnancy]. AB - The authors aimed to assess the impact of a routine ultrasound examination performed between 10 and 13 weeks of pregnancy. During a thirty month period, 778 ultrasound examinations between 10 and 13 weeks of pregnancy were performed, in women referred to our hospital. Transvaginal ultrasound was systematically adopted and the parameters obtained were introduced in a computerized data base. Biographic data, first day of menses (whenever possible), menstrual cycle characteristics, eventual use of hormonal contraception in the three months before last menses, antecedents of chromosomal abnormalities, number of foetuses and chorionicity, foetal vitality, crown-rump length, nuchal translucency and foetal heart rate were registered in all examinations. The median gestational age at the time of examination was 12.5 weeks (9-14.3). The median of maternal age was 29 years (14-44), maternal age prevalence higher or equal to 35 years was 17%. Fifty two per cent of women had usually regular menstrual cycles and 11% ignored last menses. In 74% of cases discrepancy between amenorrhea and ultrasound derived gestational age was inferior to one week and in 19% superior. The median of nuchal translucency was 1.4 mm (0.5-10), 7% of all cases had a nuchal translucency higher or equal to 2.5 mm. If maternal age criteria had been decisive for diagnostic invasive procedures, they would have been made in 135 cases. Considering nuchal translucency value combined with maternal age, it should have been done in 63 cases. In our series, invasive testing was performed in 31 (5%) cases. Eight women with fetuses with abnormal karyotypes decided for termination of pregnancy. The importance of ultrasound examination between 10 and 13 weeks seems unquestionable, allowing the correction of gestational age, multiple pregnancy characterisation and chromosomal abnormalities screening. PMID- 12645217 TI - [Smoking habit cessation in coronary patients]. AB - OBJECTIVE: Assess the impact of coronary events in smoking cessation habits, and study the association between smoking habits and coronary artery disease. METHODS: With a clinical structured interview, three groups were selected (n = 68), from the patients of the Cardiology Service of Hospital Militar Principal. Sample 1 was composed by patients with stable, and unstable angina (n = 18), sample 2 by patients with myocardial infarction (n = 26), and sample 3 by individuals with other cardiac disease (arterial hypertension, or valve disease) (n = 24). The individuals were aged between 48 and 75. The following statistical procedures were used: correspondence analysis, chi-square test (chi 2), and Fisher exact test. RESULTS: The association between smoking habits (non smoker, former smoker, and current smoker), and type of disease (angina, myocardial infarction, and arterial hypertension), was statistically significant [chi 2 (68) = 14.5; p = 0.006], and there was also a significant association between smoking habits and coronary artery disease [chi 2 (68) = 4.0; p = 0.045]. The category of ex-smokers and myocardial patients also showed significant association (Fisher exact test = 0.036). CONCLUSIONS: Previous myocardial infarction events constitute an experience of severe suffering and crisis, in which the patient accepts to stop smoking. We also confirm that smoking is a risk factor for myocardial infarction. Thus the objective of clinic interventions, aiming to promote the smoking cessation intentions, are important to promote this decision. PMID- 12645219 TI - [Multiple organ resections in surgery of the colon and rectum]. AB - Colorectal cancer patients with regional invasion or metastatic disease present low life expectancy. Radical surgical treatment is the only potentially curative procedure, with significant good results being reported. Thirty two patients with either regional invasion or metastatic disease were submitted to surgery with good survival rate. PMID- 12645220 TI - [Neuroendoscopy in the treatment of obstructive hydrocephaly]. AB - OBJECTIVE: The purpose of this study was to determine the safety and efficacy in the first sixteen patients operated by endoscopic third ventriculostomy for triventricular obstructive hydrocephalus in the Hospital of S. Joao-Porto. METHODS: Operated from December 1998 to December 1999, there were one adult, one teenager and 14 children, with a average age of 18 months in the paediatric group. The three major causes for the changed CSF dynamics in this 16 patients were: three had aqueductal congenital stenosis; nine had aqueductal acquired obstruction (three with post-infectious occlusion, three with tumours, two with intraventricular cysts and one with a pineal arteriovenous malformation), and four with obstruction of the basal arachnoidal cisterns associated with myelomeningocele. RESULTS: The endoscopic third ventriculostomy (ETV) was concluded in 15 of the selected cases (it was not possible in a new-born with hydrocephalus after bacterial meningitis), and it was repeated in two patients. The success rate (clinical efficacy of the procedure) was 9/15 (60%), and the mean follow-up was 9.1 months (range from three to 15 months). CONCLUSION: ETV is a efficient method in the treatment of non-communicant hydrocephalus. The safety and low-time consuming of this technique, avoiding the shunt disease, makes it desirable specially in paediatrics patients, or even in the case of different cases of hydrocephalus were some obstructive factor could be identified. So, some patients with a previous shunt or with a diagnosis of myelomeningocele, can benefit with this procedure. PMID- 12645221 TI - [Intestinal duplication. A diagnostic and therapeutic challenge]. AB - Duplications of the alimentary tract are rare congenital anomalies. The symptoms often mimic other surgical diseases and the diagnostic workup presents low accuracy. The charts of twelve children with intestinal duplications were reviewed. Their ages ranged from 13 days to 12 years (median: 12 months). The majority of the cases were located at ileum (n = 9). These children presented: rectal bleeding (n = 4), intussusception (n = 3), intestinal obstruction (n = 1) and hemoperitoneum (n = 1). All that were symptomatic before 1 year of age presented with intussusception (n = 3/3). All, except one colonic case was cystic. 99mTc abdominal scan was positive in 3 cases presenting rectal bleeding. One child with gastric duplication was diagnosed prenatally. Treatment consisted of enterectomy in all but one case, with no complications. Experience in the diagnostic workup and management is important to achieve a low morbidity rate in this congenital anomaly, which may present delayed manifestations. PMID- 12645222 TI - [Urolithiasis and renal colic. Therapeutic approach in urology]. AB - Renal colic is one of the most common pathologies found in our emergency services. Although a source of great pain and apprehension, most of the episodes do not need a specialised approach. The authors present a general overview in the proposed management of kidney lithiasis, including both recommendations for a conservative, expectant or prophylactic treatment, and recommendations and different techniques for urological intervention. PMID- 12645223 TI - [Malignant peritoneal mesothelioma -- diagnostic and therapeutic difficulties]. AB - Malignant Peritoneal Mesothelioma (MPM) is an uncommon tumour. Clinical presentation is indefinite and differential diagnosis with peritoneal carcinomatosis is difficult. It has a bad prognosis (survival of 6-12 months) due to the delay of the diagnosis and inefficacy of the conventional therapeutic. The authors present a case of MPM which was diagnosed over the investigation of ascitis. The laparoscopic peritoneal biopsy permitted the diagnosis. After systemic quimiotherapy, the patient is in partial remission at 42 months of follow-up. PMID- 12645224 TI - [Cutaneous leishmaniasis. Unusual disease in Portugal?]. AB - Due to its morbility and disfiguring scars, cutaneous leishmaniasis is a serious Public Health problem. The incidence of the disease, world wide, is estimated to be of 1 million to 1.5 million cases per year. In Portugal, it has been considered a rare disease and sporadic cases have been described. In Southwest Europe, Leishmania infantum has been the only agent responsible for the autochthonous cases of cutaneous leishmaniasis. In 1994, in an portuguese clinical case, the parasite isolated from a cutaneous lesion was identified as L. infantum zymodeme MON-29. As described in this work cutaneous leishmaniasis should not be considered, any longer, a rare disease in Portugal. PMID- 12645225 TI - Ethical issues at the university-industry interface: a way forward? AB - This paper forms an introduction to this issue, the contents of which arose directly or indirectly from a conference in May 2001 on Corruption of scientific integrity?--The commercialisation of academic science. The introduction, in recent decades, of business culture and values into universities and research institutions is incompatible with the openness which scientific and all academic pursuit traditionally require. It has given rise to a web of problems over intellectual property and conflict of interest which has even led to corporate sponsors' suppressing unfavourable results of clinical trials, to the detriment of patients' health. Although there are those who see the norms of science developing to recognise the importance of instrumental science aiming at specific goals and of knowledge judged by its value in a context of application, none justifies the covert manipulation of results by vested interest. Public awareness of these problems is growing and creating a climate of opinion where they may be addressed. We suggest a way forward by the introduction of nationally and internationally-accepted guidelines for industrial collaboration which contain proper protections of the core purposes of universities and of the independence of their research. Some codes suggested for this purpose are discussed. We note that some universities are moving to adopt such codes of conduct, but argue the need for strong support from the government through its funding bodies. PMID- 12645226 TI - Non-instrumental roles of science. AB - Nowadays, science is treated an instrument of policy, serving the material interests of government and commerce. Traditionally, however, it also has important non-instrumental social functions, such as the creation of critical scenarios and world pictures, the stimulation of rational attitudes, and the production of enlightened practitioners and independent experts. The transition from academic to 'post-academic' science threatens the performance of these functions, which are inconsistent with strictly instrumental modes of knowledge production. In particular, expert objectivity is negated by entanglement with political and commercial interests. We cannot go back to the old academic model for science, but need to consider how to maintain its vital non-instrumental roles. PMID- 12645228 TI - Problems for biomedical research at the academia-industrial interface. AB - Throughout much of the world, universities have driven towards industrial partnerships. This collaboration, which, in the biochemical field at least, has to continue if potential benefits for patients are to be realised, has brought with it a number of problems. These include the neglect of long-term research in favour of short-term projects, the curtailing of free dissemination of research information within university departments and the biasing of results of clinical trials by the financial interests of the investigators. It is very important that governments, universities, and industry itself address these problems. Universities should monitor the amount of basic, curiosity-driven research that is being carried on, compared with that which is more short-term goal orientated. PhD students and post-doctoral fellows should be exposed to the principles of bioethics early on in their careers. Further work is necessary on the terms of research contracts to protect, on the one hand, the rights of individual scientists and, on the other, industry from rogue scientists. Where problems arise, procedures should be in place for independent reviews to be conducted by bodies such as the Medical Research Council in the UK or the National Institutes of Health in the USA. The conflict-of-interest rules recently introduced for publication in medical journals should be extended to all branches of science. PMID- 12645227 TI - Patients' health or company profits? The commercialisation of academic research. AB - This paper is a personal account of the events associated with the author's work at the University of Toronto's Hospital for Sick Children on a drug, deferiprone, for the treatment of thalassaemia. Trials of the drug were sponsored by the Canadian Medical Research Council and a drug company which would have been able, had the trials been successful, to seek regulatory approval to market the drug. When evidence emerged that deferiprone might be inadequately effective in a substantial proportion of patients, the drug company issued legal threats when the author proposed informing her patients and the scientific community. Until protests were made by international authorities in her field of research, the hospital and university did not adequately support the author's academic freedom and responsibilities as a medical practitioner. It is argued that underlying cause of this, and of other similar cases, is the political philosophy which is driving the commercialisation of universities and bringing about the deregulation of drug approval procedures. Together these changes constitute a serious threat to the public good. PMID- 12645229 TI - Guard dogs of perception: the corporate takeover of science. AB - The belief that science is "a driver of growth in the knowledge economy" has led in recent decades to increasing encouragement by government of the involvement of industry and of commerce in the sponsorship and direction of research in universities, and to the increasing influence of industrial representatives on advisory panels associated with the publicly funded scientific research councils. By extending the doctrine of commercial confidentiality into university laboratories, inconvenient findings have been suppressed, and both free endeavour and free speech undermined. This has narrowed our scientific horizons and compromised government advisors. It is argued that scientific freedom is a guarantor of our wider liberties. Science, which tells us who we are and how we can live better, is being distorted so twisting our understanding of the ways in which we might progress, shutting off alternatives to existing models of development. Business now stands as a guard dog at the gates of perception. Only the inquiries which suit its needs are allowed to pass. PMID- 12645231 TI - The ethical implications of the new research paradigm. AB - Research is now an increasingly heterogeneous activity involving an expanded range of new actors and stake-holders and employing an eclectic range of epistemologies and methodologies. The emergence of these new research paradigms- and, in particular, of so-called 'Mode 2' knowledge production that is highly contextualised and socially distributed--raises new and challenging ethical issues and also important questions about the autonomy of science and the social responsibilities of scientists. PMID- 12645230 TI - In the grip of the python: conflicts at the university-industry interface. AB - When the University of Toronto withdrew a contract it held with me in December 2000, it initiated a sequence of events that led to a public letter to the University from senior figures in the world psychopharmacology community protesting against the infringement of academic freedom involved and a first ever legal action, undertake by this author, seeking redress for a violation of academic freedom. The issues of academic freedom surrounding this case have been intertwined with a debate about the possibility that the selective serotonin reuptake inhibitor (SSRI) group of antidepressants have the potential to trigger suicidality in a subgroup of patients. Whether the SSRIs do trigger suicidality or not, exploration of this issue has given rise to a number of worrying sets of observations. First, in my view, there is evidence that pharmaceutical companies have miscoded raw data on suicidal acts and suicidal ideation. Second, this author also maintains that there is a growing body of examples of ghostwriting of articles in the therapeutics domain. Many of the tensions evident in this case, therefore, can be linked to company abilities to keep clinical trial data out of the public domain--this is the point at which the pharmaceutical python gets a grip on academia. PMID- 12645232 TI - G.A.T.S. and universities: implications for research. AB - The likely impact of applying the General Agreement on Trade in Services (GATS) to higher education are examined. GATS aims to "open up" services to competition: no preference can be shown to national or government providers. The consequences for teaching are likely to be that private companies, with degree-awarding powers, would be eligible for the same subsidies as public providers. Appealing to the inadequate recently introduced "benchmark" statements as proof of quality, they would provide a "bare bones" service at lower cost. Public subsidies would go: education being reduced to that minimum which could be packaged in terms of verifiable "learning outcomes". The loss of "higher" aspirations, such education of critically-minded citizens of a democratic and civilized society would impoverish the university's research culture which demands honesty and openness to public scrutiny. Most university research is substantially supported by public subsidy. Under GATS discipline, commercial providers of research services could be entitled to similar public subsidies. Publicly funded fundamental research would fade, leaving university research totally dependent for funds upon the good will of industry and commerce. Present problems, such as the suppression of unwelcome results and the use of questionable results to manipulate public opinion, would considerably increase. The public would lose a prime source of trustworthy knowledge, needed in political discourse, legal disputation, consumer protection and in many other contexts. PMID- 12645233 TI - Conflict of interest in biomedical research: a view from Europe. AB - In this paper I address the conflict of interest (CoI) issue from a legal point of view at a European level. We will see that the regulatory framework that exists in Europe does state the need for the independence of ethics committee involved in authorisation of research and clinical trials. We will see that CoI is an element that has to be closely monitored at National and International level. Therefore, Member States and Newly Associated States do have to address CoI in the authorisation process of research and clinical protocols of biomedicine. PMID- 12645234 TI - Masked ball: ethics, laws and financial contradictions in Hungarian health care. AB - Corruption is a major problem in the societies of the post-communist Central European countries. Corruption in health care has some unique characteristics undermining the efficacy of and respect for Hungarian health care. One of the forms of corruption is tipping. This highly contested phenomenon is present in most of the patient/health professional's interactions in a sophisticated manner, raising serious ethical and legal dilemmas. The present paper analyzes tipping and other corruption-related factors, such as financial conflict of interest between industry and health care and argues that since ethical and legal considerations are often ignored in the country, patient care and clinical research are affected by these controversial issues to a great extent. PMID- 12645236 TI - GASB proposes new standards for financial reporting of postemployment benefits by state and local governments. AB - GASB has proposed new standards that will affect the way in which governments report postemployment health care benefits in audited external financial statements, resulting in more complete and transparent reporting by employers and plans and more relevant and useful information for the users of governmental financial reports. This article provides an overview of current financial reporting standards and practice, the financial reporting objectives of the project, the proposed measurement approach, noteworthy specific proposals, and the projected timetable for completion of the project and implementation of the new standards. PMID- 12645237 TI - Dealing with the prescription drug dilemma. AB - When the West Virginia Public Employees Insurance Agency saw 20% increases in drug spend, they formed a coalition with several other states and negotiated a contract with a new PBM that may save $25 million over three years. Other initiatives include a generic copayment waiver and a counter-detailing program. PMID- 12645235 TI - The Olivieri Report--a compelling study of the growing tensions in clinical research. PMID- 12645238 TI - Designing your health benefit plan. AB - A health care benefit plan should meet the needs and expectations of your organization and its insureds. Each organization is unique, so a health care benefit plan shouldn't be "off the shelf" but be uniquely tailored for your organization. Analyze current demographic, utilization and other data to determine which services members and their families are using and what type of services may be needed in the future. PMID- 12645239 TI - COBRA compliance: how employers can successfully meet today's complexities. AB - Although the architects of COBRA had sound and compassionate motivations in place, administration of and compliance with this law are far from easy. COBRA assists employees that lose their jobs by allowing them to purchase insurance benefits from their former employer. Outsourcing COBRA administration can be the best way for some employers to cope with COBRA regulations, contingencies and paperwork and avoid legal fees and penalties. But look for COBRA providers that have a sound track record. PMID- 12645240 TI - HIPAA privacy rules and compliance with federal and state employment laws: the participant authorization form. AB - Although HIPAA privacy rules do not apply directly to employers or to employment records, they do apply indirectly to employers because employer-sponsored group health plans and all health care providers are covered. HIPAA privacy regulations overlap with federal and state employment laws, and liabilities for breach of confidentiality can be substantial. Employers can use a participant's authorization form to comply with employment laws and company policies. Sample authorization forms are provided at the end of this article. PMID- 12645241 TI - Protecting the privacy of personal information in employee benefit plans. AB - The United States is not the only country dealing with privacy of health information. In the absence of clear legislative direction, this paper focuses on the common privacy principles that underlie Canadian privacy legislation and recommends a plan of action. The objective is to give plan administrators and trustees sufficient transition time to develop and implement the appropriate privacy strategies, thereby minimizing both the potential burden of compliance and the potential risks of noncompliance. PMID- 12645242 TI - Liability of consulting physician. PMID- 12645243 TI - Cause for alarm? Predictions say physician shortage is fast approaching. PMID- 12645244 TI - IPAs give MDs a leg up. PMID- 12645245 TI - Prevention partners. Community, physicians team up against injury and violence. PMID- 12645246 TI - The alliance tackles a new public health crisis: bullying. PMID- 12645247 TI - Liability of consulting physician. PMID- 12645248 TI - Reimbursement and liability issues. Top specialty societies' agendas. PMID- 12645249 TI - Michigan's liability climate holds steady during tumultuous times. PMID- 12645250 TI - In a strained system, there remain options for charity care. PMID- 12645251 TI - Oral health and overall health. The mouth can tell the tale. PMID- 12645252 TI - Science finds no link between MMR vaccine and autism. PMID- 12645253 TI - It's time to get ready for the coming flu season! PMID- 12645254 TI - [Studies of mutations in BRCA1 transactivation domain by visualization of chromatin structure]. AB - Mutations in breast cancer susceptibility gene 1(BRCA1) account for approximately 40%-50% of familial breast cancer cases and for more than 80% of inherited breast and ovarian cancer cases. Many cancer-predisposing mutations are located in the C terminal region that functions as a transcriptional activation domain, but most of the mutations in the transactivation domain identified to date cannot be readily distinguished as either disease-associated mutations or benign polymorphisms. Because chromatin structure regulation is an early event in gene transcription control, the chromatin unfolding activities of different transactivation domain variants were compared with that of the wild-type transactivation domain by use of an approach that allows visualization of large scale chromatin structure through lac repressor/lac operator recognition. To do this, different constructs of the transactivation domain were selected as follows: (a) the wild-type transactivation domain; (b) two polymorphisms (S1613G and M1652I); and (c) four cancer-predisposing mutations (A1708E, M1775R, W1837R and Y1853 term). All of the constructs were made by fusing in frame with lac repressor. Western blot analysis indicated that all of the fusion proteins were expressed in A03 1 cells, in which multiple copies of the lac operator were integrated to produce a heterochromatic region of the genome. The chromatin unfolding assay showed that, like the wild-type transactivation domain, two variants that represent benign polymorphisms did not induce chromatin unfolding or only induced subtle change. Contrary to the behaviors of the wild type and two benign variants, four cancer-predisposing mutations in the transactivation domain superactivate the chromatin unfolding. The results suggest that the chromatin unfolding assay can aid in the characterization of deterious mutations in the C terminal transactivation domain of BRCA1 and may provide more reliable presymptomatic risk assessment. PMID- 12645255 TI - [Study of ABO blood group secretor type alpha(1,2)-fucosyltransferase gene polymorphism in Chinese]. AB - Secretor gene (FUT2)-encoded secretor type alpha(1,2)-fucosyltransferase (Se enzyme) that regulates expression and secretion of ABO(H) antigens in epithelial cell of glands and body fluids. It has the extensive polymorphism and race specificity. In the present study, we investigated the distribution of the fusion gene of the FUT2 locus in 191 Manchu individuals from Liaoning Province and 208 Mongolian individuals from Inner Mongolia, and analyzed the polymorphism of the FUT2 gene in 90 unrelated Han Chinese from Shandong Province and 90 Mongolian from Inner Mongolia, respectively. The fusion gene was not found in the two investigated ethnic groups. The frequencies of G849A nonsense mutation in Shandong Han Chinese and Mongolian of Inner Mongolia individuals were the same, 0.0055. PMID- 12645257 TI - [Studies of STR polymorphisms in Pumi and Lisu minorities in China]. AB - In this study, we investigated the polymorphisms of STR of Pumi and Lisu minorities with STR genescan marked by fluorescence. Eighty-five alleles of 9 STR in Pumi were detected with the frequency 0.0050-0.5250 and 194 genotypes were found with frequency of 0.0098-0.3235. Sixty three alleles with their frequency of 0.0050-0.4802 and 145 genotypes were found out with frequency of 0.0099-0.3664 in Lisu population. Hi-Square test indicated the distribution of genotypes agreed with Hardy-Weinberg equilibrium (P > 0.05). Statistical analysis showed the followings: H > 0.6 in each locus, the average PIC > 0.7, mean DP > 0.8, EPP > 0.5, indicating the STR markers used in the study were of great value in the researches on minority genetics. PMID- 12645256 TI - [LIM protein KyoT2 interacts with human tight junction protein ZO-2-i3]. AB - It was reported that LIM protein KyoT2 negatively regulated transcription by association with the RBP-J DNA-binding protein. Using yeast two-hybrid system with LIM protein KyoT2 as a bait, we have isolated an alternatively spliced form of human tight junction protein 2--ZO-2-i3. Sequence analysis indicated that ZO-2 i3 is composed of 19 exons, and selected usage of exons led to an alteration in the region following the kinase domain as compared with the published sequence. To identify the interaction between LIM protein KyoT2 and ZO-2-i3, yeast two hybrid system, purification of KyoT2 protein, and GST pull-down assay were performed in the experiments. After KyoT2 and ZO-2-i3 changed vectors, positive two-hybrid yeast was obtained. Using KyoT2 protein and antibody in GST pull-down assay positive result was also obtained. Therefore we conformed KyoT2 interacted ZO-2-i3 in vitro. Furthermore it was identified in yeast that KyoT2 associated with ZO-2-i3 through its LIM2 domain. PMID- 12645258 TI - Comparative allozyme analysis of Oriental migratory locust Locusta migratoria manilensis from two breeding areas in north China. AB - The allozyme analysis using horizontal starch gel electrophoresis was employed to compare the genetic structure in the population of oriental migratory locust Locusta migratoria manilensis from two breeding areas, Beidagang(Tianjin) and Huanghua(Hebei). The two areas are adjacent but with distinct ecological features, with the recorded locust outbreaks and migration. The zymograms showed that among nineteen loci four (Mdh-1, Pgm, Adk and G3pd) showed extremely low variability level with the frequency of the most common allele higher than 0.95 in the populations from both sites. The rest loci had 2 to 4 alleles but the allele frequencies between the two populations were all similar except Fbp and Got-2 loci. In the 27 chi 2-tests for the genotypes at polymorphic loci only two (Pgi and Got-1) of beidagang population did fit the Hardy-Weinberg's expectations. This is due to high frequencies of the most common homozygotes and the corresponding heterozygote deficiency. The allozyme data demonstrated that the locusts had remarkable genetic variability within each population, but little divergence between the populations. The genetic variability measurements were found similar: Percentage of polymorphic loci (P) was between 73.7% and 78.9%; The mean number of alleles per locus (A) was from 2.9 to 3.1; and the mean heterozygosity (Ho) was nearly identical (about 0.138). The F-statistics (FST = 0.053) also showed the genetic uniformity of the populations, corresponding to the high Nei's genetic identity (I = 0.938). These results of the allozyme analysis suggested that the two populations appeared to be a part of a large population. It is reasoned that the genetic polymorphism and differentiation at certain loci between the two populations may depend on at least two agnostic factors that are all related to migration. First, the unusual dispersal capability of L. m. manilensis tends to make a continuous genetic structure distribution. Second, the frequent migration also results in the individuals to be exposed to drastically various environments. Since the broad adaptability is crucial to survive the changing environments, the genetic variation at population level is necessarily required to offer the population resilience for successful survival and reproduction under those ecologically divergent abiotic and biotic conditions. Thus, the migration contributes to the maintenance of dynamic equilibrium of genetic polymorphism in this highly specialized subspecies. PMID- 12645259 TI - [The construction of the genetic map and QTL locating analysis on chromosome 2 in swine]. AB - The study constructed the genetic linkage map of porcine chromosome 2 and further analysis of quantitative trait loci was conducted. The results of the study demonstrated that all 7 microsatellite loci we chose were with relatively high polymorphism, and its polymorphic information content was from 0.40182 to 0.58477. The genetic map we constructed for resource family was 152.9 cM in length, with the order of all loci highly consistent with the USDA map. All marker intervals were longer than USDA map with the interval between marker Sw2516 and Sw1201 as an exception. Furthermore, we conducted QTLs locating analysis by combining the genetic map with the phenotypic data. QTLs affecting lively estimated traits such as lean meat percentage, were located at 60-65 cM on chromosome 2, while QTLs for the height and marbling of Longissmus dorsi muscle were located at 20 cM and 55 cM, respectively Among them, QTL for estimated lean meat percentage was significant at chromosome-wise level (P < 0.01) and was responsible for 21.55% of the phenotypic variance. QTLs for the height and marbling of Longissmus dorsi muscle were responsible for 10.12% and 10.97% of the phenotypic variance, respectively. The additive and dominance effect of lively estimated traits were in the inverse tendency, while the QTL for the height of Longissmus dorsi muscle had its additive and dominance effect in the same tendency and was with advantageous allele in Large White. The QTLs we detected had relatively large effect on phenotype and built a basis for molecular marker assisted selection and breeding. PMID- 12645261 TI - [A novel system for characterization of the transcription activating proteins in mammalian cells]. AB - A system used for detecting the transcriptional activating activity of the function-unknown gene products in mammalian cells was developed. Based on the plasmid pTet-Off and the eukaryotic expressing vector pCDNA3.1B(-)/myc-his, firstly, we constructed a set of recombinant plasmids namely pZHO1 (for cloning into the foreign gene fragment and as a negative control), pZHO2 (as a positive control). The system also includes the plasmids pTRE-luc (encoding the Firefly luciferase reporter gene) and pRL-TK (encoding Renilla luciferase gene as background control). To confirm the feasibility of the system, the plasmids pZHO1, pZHO2 and pZHO3 (encoding p53 transcriptional activating domain, containing 73 amino acids in its N terminal) was contransfected into such mammalian cells as C4-2, MCF-7, COS7 respectively, each with pTRE-luc and pRL-TK plasmids, the feasibility of the system was determined by comparing the relative activity of Firefly luciferase activity ratio of and Renilla in different transfecting panels. Our research result showed that the system we constructed can be used for detecting the transcriptional activating activity of the target protein molecules in mammalian cells. PMID- 12645260 TI - [Research on Alu element inserted mutation in porcine FSH beta subunit gene]. AB - The inserted fragment of FSH beta subunit gene in Laiwu pigs, an excellent local pig breed in North China, Duli pigs and Landrace pigs was amplified, cloned and sequenced. Sequence analysis showed that the length of the fragments inserted between +809 bp and +810 bp of the published sequences (D00621) were 275 bp, 277 bp and 274 bp in Laiwu, Duli and Landrace pigs, respectively. And the poly (A)s in these inserted fragments were 17, 19 and 16 adenines, respectively. They were all shorter than those in Taihu pigs(292 bp and 32 adenines, respectively) reported formerly. According to the RNA polymerase III promoter structure and Alu I restriction enzyme site in the inserted fragment, it should be regarded as an Alu element. FSH beta subunit gene was considered as one candidate gene of pig litter size trait, while RNA polymerase III promoter could promote the transcription of the neighboring chromosome sequence so as to control the expression of FSH beta subunit gene or other genes. As a result, the major difference of the inserted fragment among different pig breeds was originated from the length of poly (A) end. So it was assumed that the poly (A) structure in the inserted fragment could influence the pig litter size. Because Laiwu pigs with genotype AA had 1.2 litter size more than those with genotype BB, it could be concluded that FSH beta subunit gene was related with pig litter size traits or linked with the genes of which, and the poly(A) structure in this Alu element played a critical role. PMID- 12645263 TI - [Molecular mapping of quantitative trait loci (QTL) for characters of vascular bundles in peduncle related to indica-japonica differentiation in rice (Oryza sativa L.)]. AB - A doubled-haploid population, consisting of 81 DH lines derived from the F1 hybrid of a cross between an indica cultivar and a japonica cultivar, was used to map quantitative trait loci (QTL) controlling numbers of vascular bundles in peduncle, primary rachis branches and the ratio of vascular bundles to primary rachis branches (V/R ratio). For vascular bundles, three QTL were detected. Among them, the QTL qVB-8 with the largest effect individually accounted for 31.1% of the total variation. Two QTL controlling primary rachis branches were identified and they were co-located with 2 of the 3 QTL for vascular bundles respectively. Three QTL for the V/R ratio were mapped on chromosome 1, 2 and 8, respectively. Of the three QTL, the QTL qV/R-1 with the largest additive effect, explained 25.3% of the total variation, was located on chromosome 1 and found to be closely linked to the gene sh-2, a major gene underlying grain-shattering ability. In addition, four and two pairs of significant epistatic QTL were detected for vascular bundles and the V/R ratio, respectively, but none for rachis branches. Our results suggested that the numbers of vascular bundles and primary rachis branches were independently controlled by different polygenic systems, but the two polygenic systems shared a fraction of quantitative trait loci. It was also shown that the chromosome region carrying the QTL qV/R-1 on chromosome 1 might play an important role in the processes of indica-japonica differentiation in rice. PMID- 12645262 TI - [Phylogenic analysis of the Sox gene family of vertebrate]. AB - Sox genes of vertebrate are highly evolutionarily conserved, which encode different transcriptional factors involved in various developmental processes. Sox family is characterized by a sequence-specific DNA binding HMG-box containing about 79 amino acids. To realize the complexity of genes of the Sox family in the structures, functions and their evolutionary relationships, in the present study by utilizing all available complete nucleotide/protein sequence data of vertebrate Sox genes, we performed multi-sequence comparison and construction of phylogenic tree, and the grouping of Sox family members and the pattern of their molecular evolution was also analyzed. PMID- 12645264 TI - [Molecular mapping of the fertility restorer gene Rf-4 for WA cytoplasmic male sterility in rice]. AB - The cytoplasmic male sterility for wild-abortive (CMS-WA) has been wildly used for hybrid rice breeding in China. The fertility restoration of CMS-WA is controlled mainly by two independent and dominant nuclear fertility restoring genes, Rf-3 and Rf-4. To map the Rf-4 gene with molecular markers, rice YAC clones of RGP, Japan were used to create new molecular marker. YAC contigs located between RFLP markers R1877 and G2155 on chromosome 10 were confirmed by hybridization with 12 RFLP probes. Six YAC clones, Y4630, Y2670, Y4892, Y2111, Y3821 and Y5528 were identified. Chromosome DNAs of the YAC clones were prepared and separated by CHEF. A total of 119 probes were created by sub-cloning of the YAC DNAs. RFLPs were screened between Zhenshan 97A and its near-isogenic lines with Rf-4Rf-4 genotype. Two probes, Y3-8 from Y4892 and Y1-10 from Y4630, were found to be polymorphic. Using F2 populations from crosses between Zhenshan 97A and its near-isogenic lines ZSR11, Y3-8 and Y1-10 were mapped to Rf-4 locus with genetic distances of 0.9 cM and 3.2 cM, respectively. PMID- 12645265 TI - [Studies on the distorted segregation of foreign genes in transgenic rice progenies]. AB - The segregation mode of transgenes was investigated in the transgenic progenies of three rice varieties (lines) produced by Agrobacterium-mediated transformation. The transgenic lines all contained the Bacillus thuringiensis cry1Ab gene, under the control of a maize ubiquitin promoter, and linked in tandem with gusA and hpt genes. PCR analysis showed the transgenes cry1Ab and gusA co-segregated in all self and crossed progenies tested. Therefore, GUS bio assay of leaf or endosperm tissues was used to monitor transgene segregation in the experiment. It was found that the ratio of positive to negative plants was significantly smaller than 3:1 in all heterozygous plants derived progenies, which implied the segregation biased from typical Mendelian mode for a single dominant gene. Less GUS positive plants, and consequently less homozygous transgenic lines than expected were recovered from the self progenies. Transgenic heterozygous plants (+/-) were crossed as female or male parent to conventional rice varieties (-/-), and the ratio of gusA positive (+/-) to negative (-/-) plants was investigated in test F1 population. When used as female parent, the segregation fit to 1:1, but significantly smaller than 1:1 when used as male parent. The seed-set of transgenic Nipponbare progeny was investigated individually for GUS positive and negative plants. It was found that the positive plants had an average seed-set of 64.5%, significantly lower than that of negative plants (77.9%). The biological and genetic basis of distorted segregation of transgenes was discussed and deduced on the basis of above results, and the authors are inclined to ascribe these phenomena to the poor competitive ability of pollens carrying transgenes. PMID- 12645266 TI - [In silico cloning of glucose-6-phosphate dehydrogenase cDNA from rice (Oryza sativa L.)]. AB - In silico cloning was a new strategy of gene cloning developed with the development of genome, EST projects and bioinformatics. Using wheat glucose-6 phosphate dehydrogenase cDNA (clone: Tagpdl) sequence as a querying probe, one highly homologous BAC clone sequence was obtained from rice sequence database of GenBank and the putative cDNA sequence of rice glucose-6-phosphate dehydrogenase was assembled according to the wheat clone. Furthermore, the full-length cDNA of rice glucose-6-phosphate dehydrogenase was cloned by RT-PCR with two primers designed based on this assembled cDNA sequence. Since this fragment contained a complete ORF of 1515 bp with a stop codon in its upstream and poly(A) signal in its downstream, it could be concluded that a full-length gene (GenBank accession number AY078072), which was named as OsG6PDH. Homology analysis of OsG6PDH showed a 88% identity with wheat and the deduced amino acid showed 89%, 79% and 80% homology with G6PDH from wheat, tomato and tobacco respectively. OsG6PDH was expressed in inflorescence, embryo, root and leaf of rice, with a slightly higher in inflorescence and root. It was also discussed in this paper that the application of in silico cloning in the isolation of functional genes from rice. PMID- 12645267 TI - [Function analysis of a wheat phosphate transporter in yeast mutant]. AB - Phosphorus is a major nutrient acquired by plants and phosphorus availability is considered one of the major growth-limiting factors for plants in many natural ecosystems. For a better understanding of the function of the wheat phosphorus transporter gene named TaPT2, we did Southern blot analysis and studied its function with complementation test in yeast mutant strain MB192. Southern blot indicated that TaPT2 gene is a low-copy member and has several different members in wheat genome. In the function complementation study, TaPT2 has a similar function as PHO84 which is the phosphate transporter in yeast Saccharomyces cerevisiae. Both TaPT2 and PHO84 are able to complement the PHO84 mutant phenotype of yeast strain MB192 which lacks the phosphate transporter activity. Therefor, this assessment presents evidence that TaPT2 gene plays an important role in Pi acquisition. PMID- 12645268 TI - [RAPD analysis for the genetic diversity of Brassica rapa in Tibet]. AB - Tibet, a most beautiful place, locating in southwestern China. She has been called as the third pole of the earth. Unique geological history, complex land surface and climatic zones, various soil types, all different wild vegetations etc., all of these make Tibet a very typical area of vertical agricultural ecosystem. The ecosystem in Tibet may be the most complex in the world, which varies from place to place. Genetic differentiation of 107 accessions of Brassica rapa from Tibet plateau was studied by DNA PAPD analysis using 2210 bp random primers, the genetical distribution in 107 accession of Brassica rapa from Tibet plateau was found. The results are as follows: (1) Total 236 bands were produced from 107 Tibet oilseed accession of B. rapa germaplasm resource in Tibet, of 210 bands amplified from B. rapa germaplasm resource showed polymorphism, with the ratio 88.98%. The result showed that oilseed accession of B. rapa in Tibet has richer genetic diversity; (2) Dendrogram constructed from DNA RAPDs showed that 107 accessions of B. rapa from Tibet plateau were divided into 11 cluster by calculating genetic distance, the cluster analysis showed that the genetic variation among oilseed accessions of B. rapa was closely related with their eco geographic distribution; extensive variation existed among the accessions from Tibet Province. Based on the analysis of unique geological history, complex land surface and climatic zones, various soil types, complex growing environments, long agricultural history, different cropping systems, and natural and artificial selection as well as plant geography, plant evolution theory, it concludes that Tibet is one of the oil seed gene centers in the word. PMID- 12645269 TI - [Cloning and characterization of a homologous gene of plant class V chitinase from balsampear, Momordica charantia Linn]. AB - Balsampear (M. charantia Linn.) is a vegetable crop, highly resistant to pathogens. Chitinases were proposed to play an important role in the defense response of this crop. Based on the N-terminal sequence of a purified balsampear chitinase, a fragment (ChitB), similar to the tobacco class V chitinase gene, was amplified from the leaf RNA using 3'RACE, and the corresponding 5' sequence was further amplified by the Y-RACE method. By joining the two amplified fragments, the full-length cDNA of M. charatica homologous gene of plant class V chitinase (McChi5) was obtained. The 1348 bp cDNA contained an ORF of 1044 bp, which coded for a polypeptide of 347 amino acids. The deduced polypeptide had a predicted molecular weight of 38.3 kD and a pI of 5.77. Homology analysis demonstrated that, McChi5 protein, which contained a conserved domain of family 18 glycosyl hydrolyse, had the sequence similar to tobacco class V chitinases, several putative chitinases and chitinase-like proteins of Arabidopsis thiliania, and some chitinases from mammals, insects and bacteria. Southern blotting suggested that two copies of McChi5 gene and several homologous genes existed in the M. charatica genome. By RNA dot blotting analysis, expression of the McChi5 gene was detected in cotyledons, roots, stems, and leaves, and it was not induced by wounding treatment. The biological functions and the potential applications of Mochi5 gene were discussed. PMID- 12645272 TI - Fact sheets from the Surgeon General's Call to Action to Prevent and Decrease Overweight and Obesity. PMID- 12645270 TI - [Molecular evolution of beta-glucuronidase in vitro: obtaining thermotolerant GUS gene]. AB - The Escherichia coli beta-glucuronidase gene (gus) has been developed as a reporter gene for plants, and has been widely used for over a decade. Both chromogenic and fluorogenic GUS substrates have been synthesized, allowing rapid nonradioactive assays. The use of the Escherichia coli enzyme beta-glucuronidase (GUS) as a reporter in gene expression studies is limited by some plants and plant-associated bacteria express endogenous glucuronidase activities. The use of the enzyme as a reporter in transgenic plants is limited by high false positive. Laboratory evolution methods were used to enhance the thermostability and activity of the beta-glucuronidase. Using plasmid pBI121 as template, a 1.8 kb specific product was amplified and cloned into the vector pBluescript SK. The result of nucleotide sequence analysis was the same as reported. In vitro recombination (DNA shuffling), which involves DNase I digestion, primerless PCR, and primer PCR was used to generate mutant libraries. The mutant GUS3-3 gene was isolated after three rounds of mutation, DNA shuffling, and screening. The GUS3-3 enzyme can resistant high temperature up to 80 degrees C for 30 min. The nucleotide sequence analysis showed 99.2% homology between the GUS-ck gene from pBI121 and GUS3-3 gene. The deduced amino acid sequence demonstrated that 11 amino acid was changed. The Tm value of GUS3-3 is 80 degrees C and increased by 25 degrees C above GUS-ck (55 degrees C). The researches indicated the feasibility of the molecular evolution of beta-glucuronidase in vitro to improve enzymatic thermostability. PMID- 12645273 TI - Obese and overweight: a public health epidemic. PMID- 12645274 TI - Walking program creates positive results, national attention. PMID- 12645275 TI - Obesity: facts, figures and guidelines. PMID- 12645276 TI - Prescribing physical activity in primary care. AB - Physical activity is a powerful weapon for individuals interested in health maintenance. Many benefits of regular physical activity are well known to the medical community including improved weight control, healthier cardiovascular system, and tighter diabetic control. Less recognized benefits of physical activity may include stronger bones, better sleep, lower blood pressure, improved mood, opportunities for personal enjoyment, cancer prevention, and even a longer lifespan. Despite these numerous incentives, less than a quarter of the U.S. population is active enough to gain the rewards (13). West Virginians are among the least physically active; more than 40% of state residents engage in no leisure time physical activity at all (15), so it is critical that physicians in the state take a much more active role in prescribing physical activity for their patients. Although more research is needed, data exists to suggest counseling by physicians does result in increased physical activity (20-23). Attention to risk factors and specific red flags can help decide when formal pre-exercise stress testing is necessary. An organized approach includes development of the mode, frequency, duration, intensity and progression of exercise by a physician who knows the patient's interests and limitations (28). The most current consensus guidelines suggest 30 minutes of moderate-intensity physical activity daily (12). Physicians need to encourage this goal during office visits and there appears to be benefit in putting specific physical activity advice in the form of a written prescription (21). PMID- 12645277 TI - Effective dietary counseling: helping patients find and follow "the way" to eat. AB - The leading causes of mortality in the U.S. are well known: cardiovascular disease, cancer and stroke. But, in the field of preventive medicine, the argument is made that these are not the causes of death, but rather the mechanisms of death, in turn the result of true, underlying causes. Of the more than 2 million deaths that occur each year in the U.S., more than 1 million are premature and "preventable" through the modification of lifestyle and environmental exposures. Due to the epidemic of obesity and overweight, a clinical practice that neglects patient nutrition (or physical activity) patterns is neglecting the leading causes of death for patients. On this basis, routine counseling to promote healthful eating is encouraged by the U.S. Preventive Services Task Force and there is evidence that physician training in nutrition enhances counseling. PMID- 12645279 TI - The relation of obesity to cardiovascular risk factors among children: the CARDIAC project. AB - West Virginia's prevalence of obesity is among the highest in the nation, contributing to an excess mortality rate from heart disease. Individuals who are overweight and obese have a greater risk for coronary artery disease. To gain insight into the impact of obesity on other modifiable cardiovascular disease (CVD) risk factors among children, 5,887 students from 27 rural West Virginia counties participated in the school-based Coronary Artery Risk Detection in Appalachian Communities (CARDIAC) Project during the 1999-2002 school years. Results confirmed a very high prevalence of overweight and obese children in this rural, pre-adolescent population. Almost 43 percent of the children screened were considered to be overweight (BMI > or = 85th percentile), and over one-fourth of them were obese (BMI > or = 95th percentile). This high rate of obesity among schoolchildren in West Virginia is associated with increased prevalence of other CVD risk factors such as hypertension, dyslipidemia, and insulin resistance. Interventions for prevention of excess weight and obesity should be implemented through schools and community-based programs. PMID- 12645278 TI - The role of schools in preventing childhood obesity. AB - Obesity is increasing in the United States, not only among adults, but also among children and youth (1). The U.S. Centers for Disease Control and Prevention report the incidence of children at risk for being overweight or obese has doubled in the last two decades (2). Schools can play a role in preventing childhood obesity by serving healthy meals with adequate calories and nutrients, providing nutrition education that encourages healthful food selections, offering opportunities for physical activity and creating school environments that model healthful behaviors. PMID- 12645280 TI - "Where obesity goes, so goes diabetes"--dual epidemics of alarming proportions. AB - Diabetes and obesity are increasing in alarming, epidemic proportions not only in West Virginia, but also in the United States and in the world. Their combined toll encompasses not only an increased risk of premature cardiovascular disease and death, but multiple other associated co-morbidities. Recognition of their impact on healthcare systems is crucial and involves awareness and action on the part of providers at every level. Education is of paramount importance in combating these growing trends with preservation of the health and well-being of the populace as an ultimate goal. PMID- 12645281 TI - Obesity linked to some forms of cancer. AB - Researchers have found a consistent relationship between a number of diseases, including diabetes, heart disease, high blood pressure and stroke. Although study results related to cancer have been conflicting, with some showing an increased risk and others not showing such an association, obesity does appear to increase the risk of cancers of the breast, colon, prostate, endometrium, cervix, ovary, kidney and gallbladder. Studies have also found an increased risk for cancers of the liver, pancreas, rectum and esophagus. Although there are many theories about how obesity increases cancer risk, the exact mechanisms are not known. They may be different for different types of cancer. In addition, because obesity develops through a complex interaction of heredity and lifestyle factors, researchers may not be able to tell whether the obesity or something else led to the development of cancer. PMID- 12645282 TI - Contemporary surgical management of obesity. AB - The incidence of overweight and obesity is reaching epidemic proportions in the United States. The morbidity of obesity greatly influences the lives of those with this disease and upon the healthcare systems treating those afflicted with it. Currently, medical therapies are ineffective in both the prevention and treatment of obesity. Bariatric surgery has gained immense popularity over the past decade as it has been shown to be among the few means for which obesity can be successfully treated. This article reviews the current state of the art of bariatric surgery. PMID- 12645283 TI - PEIA Pathways to Wellness Program utilizes billboards, radio ads, website to say "don't super size yourself". PMID- 12645284 TI - Coalitions working to enhance obesity prevention efforts. PMID- 12645286 TI - Medicare coverage for nutrition therapy improving patient care. PMID- 12645285 TI - New obesity center at WVU offering comprehensive program.. PMID- 12645287 TI - Is your retirement at risk? PMID- 12645289 TI - Can you hear me now? PMID- 12645288 TI - Heroes of manufacturing. PMID- 12645291 TI - [Regulatory phenomena in tolerance induction]. AB - The implication of regulatory T cells in self-tolerance induction was shown in experimental models based on construction of thymic chimera and peripheric T cell transfers. The role played by the epithelial stroma of the thymus in CD4+ regulatory T cell selection was demonstrated. In the NOD (Non Obese Diabetic) strain, protection again diabetes was obtained by grafting supplementary thymuses injected with allogeneic pancreatic islets. This result suggest that the NOD thymuses are defective in the production of T regulatory cells. PMID- 12645290 TI - [Immunoregulation and autoimmunity]. AB - Peripheral tolerance to extrinsic antigens or autoantigens involves regulatory T cells, numerous but probably distinct types of which seem to exist. The investigation of their phenotype, of the cytokines on which they depend and the circumstances for their appearance allow classifying them into two categories: 1. Cells for which the appearance is observed independently of the deliberate administration of an antigen (CD25+ T cells, NKT cells, gamma/delta cells). 2. Cells which seem to intervene exclusively in response to the antigens introduced into the organism (Th2 cells, Tr1 cells). These various cells, whether related to innate or adaptive immunity, can be used to advantage as new therapeutic targets. PMID- 12645292 TI - [CD4+, CD25+ regulatory T-lymphocytes: current concepts and therapeutic potential]. AB - The subpopulation of CD4 T cells, characterized by a constitutive expression of CD25, plays a critical role in the regulation of multiple auto-immune diseases. These CD4+CD25+ T cells may act by inhibiting the activation of both CD4+ and CD8+ conventional T cells. These cells have a thymic origin and are auto reactive. They have therapeutic potentials, not only in the control of autoimmune diseases, but also in the control of allograft rejection and graft-versus-host disease. PMID- 12645293 TI - [Renal sclerosis: a public health problem]. AB - In France, 45,000 patients are treated by hemodialysis and/or transplantation for chronic renal failure. Every year 7,000 new patients need such a therapeutic approach. The estimated cost of this pathology is about 1% of the total amount of the budget of social security, even though the number of patients is limited. Some treatments were shown to be effective in improving the progression of chronic renal failure, as for example the anti-hypertensive therapy and the adequate treatment of diabetes mellitus. Collagen deposition in each segment of the kidney, mainly in the interstitium, plays a pivotal role in the progression of renal deficiency in chronic renal failure. In order to ameliorate the progression renal failure it would be essential to know: 1) the different types of collagen deposit; 2) the proteinase/antiproteinase systems involved in the remodelling of the extracellular matrix (serine-protease, metallo-protease and their inhibitors); 3) the autocrine/paracrine effects of proteases and of growth factors on collagen synthesis. The more precise the knowledge of these factors, the more useful will be new pharmacological-, gene- or cellular therapies for limiting the progression of chronic renal failure. PMID- 12645294 TI - [Inflammation, prelude to renal sclerosis: the importance of NF-kappa B]. AB - NF-kappa B comprises a family of transcription factors. These are thought to have a central role in the expression of genes involved in cell mobilization, cell proliferation and cell differentiation, and, hence, in inflammation, repair and fibrosis processes. In particular, NF-kappa B activation appears to drive a number of inflammatory diseases of the kidney and their progression to end-stage renal failure. Thus, targeting NF-kappa B activation would lead to the development of new pharmaceutical compounds that would provide novel treatment for these diseases. PMID- 12645295 TI - [Vasoactive peptides and the development of renal sclerosis: contribution of transgenes]. AB - Vasoactive peptides are implied in the development of renal sclerosis as evidenced by the efficiency of their antagonists in preventing glomerulosclerosis of experimental and human nephropathies. Genetically engineered models provide a new approach to investigate the mechanisms of the renal profibrotic actions of angiotensin II and endothelin. Overexpression of the human angiotensinogen and renin genes in rats induces renal sclerosis independently of changes in systemic hemodynamics. The same results are observed when the endothelin-1 gene is overexpressed in mice. Transgenic mice harboring the luciferase gene under the control of the collagen I-alpha 2 chain promoter (procol alpha 2[1]) and made hypertensive by induction of nitric oxide (NO) deficiency were used to study the renal profibrotic actions of vasoactive peptides. In this strain of mice, luciferase activity is an early index of renal fibrosis. Luciferase activity was increased in preglomerular arterioles and glomeruli when mice were deficient in NO. The pharmacological blockade of angiotensin II and endothelin prevented the development of renal sclerosis without modifying blood pressure. Moreover, when the endothelin receptor antagonist was administered after the development of renal fibrosis, preformed glomerulosclerosis partially regressed. Acute administration of vasoactive peptides and TGF-beta in transgenic procol alpha 2[1] mice showed that the angiotensin II activation of collagen I gene requires participation and/or cooperation of endothelin and TGF-beta. Recent data suggest that the profibrotic actions of vasoactive peptides also need the activation of EGF receptor, ERK and rho kinase pathways in renal and vascular cells. PMID- 12645296 TI - [Proteases and antiproteases in the progression of chronic renal insufficiency lesions. The role of the tissue renin-angiotensin system and the renin receptor]. AB - The role of proteases and of antiproteases in the progression of renal disease is well established. Most studies have focused on the serine-proteases of the plasmin/plasminogen activator system and on matrix metalloproteases. Recently, renin, an aspartyl-protease, has attracted much attention because of the role of angiotensin II in the progression of renal lesions and because of the discovery of a functional renin receptor. This receptor is a 45 kDa membrane-protein that binds specifically renin and prorenin. The binding of renin induces an increase of the catalytic efficiency of angiotensinogen conversion into angiotensin I by receptor-bound renin compared to renin in soluble phase, and a rapid phosphorylation of the receptor on serine and tyrosine residues associated with an activation of MAP kinases ERK1/2. Immunofluorescence and confocal analyses on normal human kidney and cardiac biopsies show that the receptor is localized within the mesangial area of glomeruli and in the sub-endothelium of kidney and coronary arteries, associated to smooth-muscle cells. In summary, this receptor exerts dual effects, mediating renin cellular response and increasing the efficiency of angiotensinogen cleavage by membrane-bound renin. These observations emphasizes the importance of angiotensin II generation at the cell surface and the cellular effects of renin add new dimensions (and complexity) to the classical dogma that angiotensin II is the only effector of the RAS. PMID- 12645298 TI - [Renal sclerosis and its treatment. Conclusions]. PMID- 12645297 TI - [The concept of nephroprotection: toward preventive and curative therapies for renal sclerosis]. AB - During the last decade, therapeutic measures based on inhibitors of the angiotensin I converting enzyme have proved to have some efficacy in the prevention of renal fibrosis. This review is devoted to new anti-fibrotic strategies aimed at inhibiting reabsorption by proximal tubule cells of filtrated proteins, preventing transdifferentiation of proximal tubule cells, inhibiting the pro-inflammatory effects of angiotensin II and endothelin, blocking TGF beta, and limiting renal ischaemia. Furthermore, experimental models and clinical reports have shown that renal fibrosis could regress, which has important therapeutic implications. The design of new preventive and curative treatments should be accompanied by the identification of urinary and serum markers of the various stages of renal fibrosis. PMID- 12645299 TI - [Plant genome sequencing: a prelude to the study of its expression]. AB - This report illustrates development of plant sequencing programmes. So far Arabidopsis genome has been completely sequenced and a draft of the rice genome is available. The Arabidopsis programmes stimulated sequencing of EST (expressed sequence tags) from numerous cultivated species thus creating an enormous resource. The major challenge is now to correctly annotate all the genes in Arabidopsis and find out a biological and biochemical function for each one. The availability of EST and genome sequence now allows one to analyse the expression of genes at the level of the whole genome. PMID- 12645300 TI - [Transcriptomes for serial analysis of gene expression]. AB - The availability of the sequences for whole genomes is changing our understanding of cell biology. Functional genomics refers to the comprehensive analysis, at the protein level (proteome) and at the mRNA level (transcriptome) of all events associated with the expression of whole sets of genes. New methods have been developed for transcriptome analysis. Serial Analysis of Gene Expression (SAGE) is based on the massive sequential analysis of short cDNA sequence tags. Each tag is derived from a defined position within a transcript. Its size (14 bp) is sufficient to identify the corresponding gene and the number of times each tag is observed provides an accurate measurement of its expression level. Since tag populations can be widely amplified without altering their relative proportions, SAGE may be performed with minute amounts of biological extract. Dealing with the mass of data generated by SAGE necessitates computer analysis. A software is required to automatically detect and count tags from sequence files. Criterias allowing to assess the quality of experimental data can be included at this stage. To identify the corresponding genes, a database is created registering all virtual tags susceptible to be observed, based on the present status of the genome knowledge. By using currently available database functions, it is easy to match experimental and virtual tags, thus generating a new database registering identified tags, together with their expression levels. As an open system, SAGE is able to reveal new, yet unknown, transcripts. Their identification will become increasingly easier with the progress of genome annotation. However, their direct characterization can be attempted, since tag information may be sufficient to design primers allowing to extend unknown sequences. A major advantage of SAGE is that, by measuring expression levels without reference to an arbitrary standard, data are definitively acquired and cumulative. All publicly available data can thus be stored in a unique database, facilitating whole-genome analysis of differential expression between cell types, normal and diseased samples, or samples with and without drug treatment. SAGE data are readily amenable to statistical comparisons, allowing to determine the level of confidence of the observed variations. A major limitation of SAGE is that, because each analysis is obligatory performed on the whole set of expressed genes, it can hardly be performed on multiple samples, for example in kinetics studies or to compare the effects of large numbers of drugs. To overcome this limitation, high-throughput detection of a subset of mRNAs is more rapidly performed by parallel hybridization of mRNAs on arrays of nucleic acids immobilized on solid supports. From this point of view, a SAGE platform is a powerful instrument for selecting the most informative subset of genes, assembling them to design microarrays dedicated to a specific problem and calibrating measurement by comparison with a standard cell model for which SAGE data are available. This approach is an attractive alternative to strategies based exclusively on pangenomic arrays. A very large amount of SAGE data are already available and the problem is now to extract their biological meaning. Knowledge on metabolic pathways is already organized so that its successful integration in a SAGE platform can be undertaken. For other cell components and pathways, the problem lies on the lack of controlled vocabulary to describe gene activities, starting form a clear definition of the concept of biological function itself. Progress in gene and cell ontology is expected to facilitate computer-based extraction of biological knowledge from existing and forthcoming SAGE data. PMID- 12645301 TI - [The prion protein]. AB - Transmissible spongiform encephalopathies form a group of fatal neurodegenerative disorders represented principally by Creutzfeldt-Jakob disease in humans, and by scrapie and bovine spongiform encephalopathy in animals. Also called prion diseases, these disorders have the property of being infectious, sporadic or genetic in origin. Although the nature of the responsible agent of these diseases is uncertain, it is clear that a protein called PrPSc has a central role in their pathology. PrPSc is a conformational variant of a normal protein called PrPC. PrPC is a glycoprotein expressed by most tissues and is attached on the cell membrane by a glycosyl-phosphatidylinositol anchor which would be consistent with roles in cell adhesion, ligand uptake, or transmembrane signaling. NMR studies revealed that the protein has a globular domain and a long amino-terminal tail that contains repeated octapeptide domains which bind metal ions with high affinities. PrPC is localized on the cell membrane in detergent resistant microdomains and may be part of functional complexes with other molecules. This is particularly relevant, knowing the possible role of the molecule in signal transduction, resistance to oxidative stress and neuronal survival. In conclusion, it appears that the understanding of the biology of PrP is essential for the understanding of the physiological function of the protein as well as for its pathological conversion considering that trafficking of this molecule governs generation of PrPSc. PMID- 12645302 TI - [Exploitation of expression profiles: examples in oncology]. AB - The analysis of biological processes has been revolutionized by the emergence of the DNA array technology. As cellular biological events are controlled by gene expression, their modulations are markers of the cellular activity. These modulations can be indicative of either a physiological process or a pathological one. Monitoring of the expression levels of thousands of genes simultaneously, the expression profiling method is based upon comparative studies where the identification of the differentially expressed genes in two samples is aimed. The two samples under study may be compared temporally or following drug treatment, they may also originate from different sources, e.g. normal versus pathological samples. In that case, gene expression profiling is conducted for diagnostics purposes or therapy monitoring, and offers an opportunity to identify new drug targets. Using different examples, we describe the potentialities of this approach in oncology. PMID- 12645303 TI - [Pharmaco-proteomic analysis: application of proteomic analysis to the discovery and development of new drugs]. AB - Pharmacoproteomics may be defined as proteomics applied to the discovery of new therapeutic targets and to the study of drug effects. Proteomics is a powerful technique for analyzing the protein expression profiles in a biological system and its modifications in response to a stimulus or according to the physiological or pathophysiological states. Thus it is a technique of choice for the discovery of new drug targets. It is also an interesting approach for the study of the mode of action of treatments and preclinical drug development. This pharmacoproteomic approach may be particularly useful for the research of new molecular alterations implicated in type 2 diabetes and/or obesity and for the further characterization of existing or new drugs. PMID- 12645304 TI - [Reproductive toxicity of lindane]. AB - The present paper bears on the main effects of lindane (gamma isomer of hexachlorocyclohexane) on endocrine and reproductive functions in mammals. This pesticide, once widely used to kill lice and a variety of pests that attack agricultural products, livestock and trees, has been progressively eliminated from many applications since the mid-1970s in Europe or USA, but is still used in the rest of the world. Lindane is absorbed through respiratory, digestive or cutaneous routes and accumulates in fat tissues. It damages human liver, kidney, neural and immune systems and induces birth defects, cancer and death. Chronic administration results in endocrine disruption in birds as well as in mammals. Treatment with 1-40 mg of lindane/kg b.w. disrupts testicular morphology, decreases spermatogenesis, inhibits testicular steroidogenesis, reduces plasma androgen concentrations and may adversely affect reproductive performances in males. In females, lindane disrupts the estrous cycle, reduces serum estrogen and progesterone levels, decreases sexual receptivity whereas in pregnant dams it decreases whelping rate and litter size. These effects were also observed in some rats exposed to residual environmental doses. In addition, there is concern that irreversible effects may be induced when animals are exposed to endocrine disrupting chemicals during critically susceptible phases of sexual differentiation or development. These effects would results from (i) alterations of gonade or gamete cell membranes (ii) cell metabolism changes including alterations of ionic exchanges (mainly calcium or potassium), direct or free radical-mediated inhibition of steroidogenesis (iii) or neuroendocrine changes leading to a decrease in sexual performance of either parents or their offsprings exposed in utero or through lactation. PMID- 12645305 TI - [Cardiotoxicity of lindane, a gamma isomer of hexachlorocyclohexane]. AB - The goal of the present review is to collect information concerning membrane effects induced by lindane intoxication, a y isomer of hexachiorocyclohexane (gamma-HCH) that has been largely used as an insecticide and disinfectant in agriculture and entered also in the composition of some lotions, creams and shampoos used against parasites (lice and scabies). Absorbed through respiratory, digestive or transcutaneous pathways, lindane accumulates within lipid rich tissues. Lindane accumulation depends on the duration of the exposure and affects tissues in the following order: adipose tissues > brain > kidney > muscle > lungs > heart > liver > blood. Whatever the mode of lindane absorption, it accumulates in blood and is distributed throughout the body. It may affect human health by exerting systemic, immunologic, teratogenic, and/or cancerogenic effects. The symptoms of lindane intoxication are different according to the mode of intoxication, acute or chronic. The absorption of high doses of gamma-HCH is particularly toxic for the central nervous system and for the female and male reproduction apparatus in mammals where lindane is considered as an endocrine disruptor. Lindane is highly lipophilic and incorporates into biological membranes according to the following sequence: mitochondria > sarcoplasmic reticulum > myelin > brain microsomes > erythrocytes. Lindane exerts a stimulating action on synaptic transmission and inhibits the chloride current activated by gamma-amino butyric acid (GABA) of many muscular and nervous preparations by interacting with the receptors GABA-chloride channel complex. It seems to affect calcium homeostasis of many tissues. The similarity between lindane and inositol (1, 4, 5) phosphate (IP3) suggested that lindane releases Ca2+ from IP3-sensitive intracellular stores in macrophages and myometrial cells. Ca2+ release from reticulum endoplasmic, mitochondria and other Ca2+ stores has been reported in cat kidney cells. Lindane altered energetic metabolism of hepatic mitochondria and the inositol-phosphate synthesis in neuronal cells. However, lindane does not compete with the IP3 receptor. Lindane produces a Ca2+ influx in mice peritoneal macrophage cells responsible for the Ca2+ induced Ca2+ release produced by phospholipase C via IP3 pathway and resulting in a maintained increase of the free cytosolic Ca2+ concentration. Lindane decreased the membrane erythrocyte and cerebral cell concentration of phosphatidyl inositol PI, PIP and PIP2 in rats repetitively exposed to lindane for 3 or 6 months. Lindane induces oxidative stress; it modifies the activity of the scavenger enzymes. This effect is involved in the inhibition of intercellular gap junctions. Modifications of the electrocardiogram (ECG), sinusal rhythm alteration and negative and dysphasic variations of T wave, similar to those produced by hyperkaliemia, have been reported after lindane absorption. During acute lindane poisoning, the activities of serum transaminases (SGOT, SGTP), and lactate deshydrogenase (LDH) increase. Lindane produces histological alterations of cardiac tissues and a cardio vascular dystrophy (contracture, degenerescence and necrosis) mainly in the left ventricular wall and a hypertrophy of the left ventricle. Chronic application of residual doses of lindane shortened the action potential duration in rat papillary muscle. These effects were similar to those induced by hyperthyroidism. Lindane increases the triiodothyronine (T3) serum level in hyperthyroid rats. T3 plays an important role in the postnatal development of the rat ventricle by increasing the density of potassium channels which contribute to action potential shortening during the development. Thyroid hormones influence the regulation and the expression of messengers ARN which encode different potassium channels involved in action potential repolarization (Kvl.2; Kvl.4; Kvl.5; Kv2.1; Kv4; HCN2). The thyrotropine-releasing hormone (TRH) modulates the HERG-type rapid delayed potassium channel (IKr) encoded by the human gene ether-a-go-go in rat anterior pituitary cells GH3/B6. This channel is involved in the cardiac long QT syndrome. TRH modifies the current kinetics of human HERG potassium channel co expressed in Xenopus oocytes with the TRH receptor, whose activity is modulated via the protein kinase C pathway linked to a G protein-coupled receptor and is regulated by changes in the PIP2 concentration in the membrane. IKr channels regulation is also dependent on sexual hormones. In conclusion, lindane affects the excitable membranes and the cardio circulatory system. These alterations (may) represent a potential risk for human health. PMID- 12645306 TI - [Nuclear pores: from yeast to higher eukaryotes]. AB - In eukaryotes, bidirectional transport of macro-molecules between the cytoplasm and the nucleus occurs through elaborate supramolecular structures embedded in the nuclear envelope, the nuclear pore complexes (NPCs), whose overall architecture has been evolutionary conserved from yeast to vertebrates. In recent years, fast progress in characterizing the NPCs components (the nucleoporins or Nups) has been made in the yeast S. cerevisiae, and to a lesser extent in vertebrates. In addition, despite the low homology between most yeast and vertebrate nucleoporins, their organization and their topological mapping within the NPC substructures have been broadly conserved during evolution. PMID- 12645307 TI - Heavy metal fish. New warnings about mercury tell seafood buyers to beware. PMID- 12645308 TI - The vaccine conundrum. Should parents be concerned about the number of childhood inoculations? PMID- 12645309 TI - Blocking colon cancer. An aspirin a day helps. PMID- 12645310 TI - A darkening sky. A smoky shroud over Asia blocks both sun and rain. PMID- 12645312 TI - You're so vain. Spas for guys and macho beauty products usher in a men's grooming boom. PMID- 12645311 TI - The globe-trotting microbe in your gut. PMID- 12645314 TI - Consumers use multiple media sources simultaneously. PMID- 12645315 TI - Health system tests CRM data base. Community Health Network uses direct mail to boost physicians. AB - A six-month pilot patient retention project for Community Health Network (CHN), Indianapolis, ran from July 2002 to January 2003. It was a direct mail campaign on behalf of some members of the group practices owned by CHN, designed to test the use of the system's CRM database. Patients of the physicians received personal, dynamically-generated cards reminding them to schedule appointments and tests. Each mailing cost $1.76, including production and mailing. PMID- 12645313 TI - Parkview opens a new hospital. Fort Wayne, Ind., hospital works with agency in complex compaign. AB - Parkview Health, Forth Wayne, Ind., recently opened its new primary care hospital, Parkview North, an expansion of the services of its flagship Parkview Hospital. But the grand opening event was only one focus of the integrated advertising campaign, which also included a component to recruit staff. In view of the nationwide shortage of nurses, marketers faced the need for 200 nurses to staff the new facility. With its agency, Ten Adams, of Evansville, Ind., Parkview succeeded in opening its new hospital with a full complement of nurses. PMID- 12645317 TI - They use humor, off-beat images. These ads keep the consumers' minds OUT of the hospital. AB - Limited budgets make it more important for a healthcare campaign to stand out. NeathawkDubuque, Richmond, Va., showcases television commercials for Washington Hospital Center and Georgetown University Hospital, both in Washington, D.C.; and print ads for the University of Virginia Health System, Charlottesville, Va. Additionally, Coil Counts Ford and Cheney, Chicago, provide a distinctive campaign for Advocate Illinois Masonic Medical Center, Chicago. PMID- 12645316 TI - John C. Lincoln Health Network recognized for community service. Phoenix institution wins prestigious Foster G. Mcgaw Prize. AB - John C. Lincoln Health Network, Phoenix, was awarded the Foster G. McGaw Prize for excellence in community service, one of the healthcare field's most prestigious honors. The network serves a broad geographic area and nearly a dozen communities. Those communities most challenged by poverty, hunger, poor housing and crime are the focus of most of the health network's efforts. PMID- 12645318 TI - Marketing goes back to the basics. It's working for Continuum Center for Health and Healing. AB - The Continuum Center for Health and Healing, affiliated with Beth Israel Hospital, New York, emphasizes education and communication--"the cornerstones of marketing integrative medicine." PMID- 12645319 TI - Hospital of Saint Raphael says it's 'where great nurses work'. Reality-based campaign brings in more nurses than hoped for. AB - Hospital of Saint Raphael, New Haven, Conn., used conventional media in an unconventional way in a campaign to recruit much-needed nurses. Their reality based campaign succeeded in adding more than 180 new nurses to the hospital staff. PMID- 12645320 TI - Hosptial's web site structured for the patient's perspective. North Memorial Medical Center site features 'care centers'. AB - Visit the redesigned web site of North Memorial Medical Center, Robbinsdale, Minn., at www.north-memorial.com. Some believe there is nothing like it in the Minneapolis/St. Paul area. Its distinction is in how its information is prioritized and presented. PMID- 12645321 TI - [Recurrent skin problems in a manatee: the diagnostic approach]. AB - A 40-year-old manatee was referred with recurrent vesicular and ulcerative dermatosis for the past 15 years. During this period the animal was anorectic and lost weight. Differential diagnoses were formulated on the basis of the history and clinical signs. Skin scrapings, bacterial and fungal culture, cytological examination, blood examination, and histopathological examination of skin biopsies narrowed this list down to autoimmune dermatosis. Despite corticosteroid therapy the symptoms recurred and the animal died. Histopathological examination of post-mortem skin biopsies showed again autoimmune dermatosis, more specifically subepidermal bullous autoimmune dermatosis, as the most probable cause of the skin lesions. Post-mortem examination showed cardiac decompensation and chronic nephritis. It was impossible to estimate the possible contribution of the chronic dermatosis to the cause of death. The purpose of this case report is to show the importance of a systematic work-up of disease in exotic animals. PMID- 12645322 TI - [Control of foot and mouth disease. Conclusions and recommendations of Working Group for Foot and Mouth Disease]. PMID- 12645323 TI - [Predecessors: veterinarians from earlier times (49). George Fleming (1833 1901)]. PMID- 12645324 TI - [Find the liver tumor]. PMID- 12645325 TI - [Veterinary midwife brought to justice]. PMID- 12645326 TI - [Via prosecutor]. PMID- 12645327 TI - [Reaction from the Society against Home Remedies]. PMID- 12645328 TI - [Veterinarians, learn to understand each other better]. PMID- 12645329 TI - [Norms and values]. PMID- 12645330 TI - [Also that!]. PMID- 12645331 TI - [What is the cause and the prophylaxis, but most of all: what is the therapy for a cow with bursitis?]. PMID- 12645332 TI - Validating immunohistochemical staining for KIT (CD117). PMID- 12645333 TI - Comprehensive graphic-based display of clinical pathology laboratory data. AB - In this age of ever-increasing demands for and uses of patient data, technologic advancements in the form of electronic patient records permit improved data access and prompt retrieval of higher quality patient care data, with more versatility in display, facilitating the integration of information concerning patients over time and between settings of care, which is in turn more accessible for use by practitioners and provides more efficient and effective decision support in areas of patient care. The graphic display of laboratory data is central to the evolving computerized patient record and needs to be taken into careful consideration along with clinician perception and ease of data interpretation in redesigning the graphic reporting of numeric clinical pathology laboratory data. An ideal system should generate user-friendly, graphic-based comprehensive reports highlighting abnormalities with trends for diagnosis, clinical management, and risk-factor detection. PMID- 12645334 TI - c-Kit expression in desmoid fibromatosis. Comparative immunohistochemical evaluation of two commercial antibodies. AB - To determine the frequency of c-Kit staining in desmoids and optimize an assay for clinical use, we stained 19 desmoids from various sites at various dilutions with 2 commonly used rabbit polyclonal, anti-c-Kit antibodies (A4502, DAKO, Carpinteria, CA; C-19, Santa Cruz Biotechnology, Santa Cruz, CA), with and without heat-induced epitope retrieval (HIER) in citrate buffer. Approdpriate external and internal control samples were evaluated for each test condition. At dilutions of 1:50 both antibodies stained substantial numbers of desmoids: with/without HIER, A4502, 89%/63%; C-19, 37%/74%. The staining was cytoplasmic without cell membrane accentuation. However, background stromal staining and nonspecific staining of endothelium and smooth and striated muscle were problematic with both antibodies at 1:50. At higher dilutions, C-19 stained no desmoid; however, diminished staining of external and internal control samples made it unreliable. A4502 similarly stained many fewer desmoids at higher dilutions. However, it retained strong staining of both external and internal control samples and showed much less nonspecific staining. Best results were achieved at 1:250 without HIER; only weak focal staining was present in 1 desmoid. With a simple immunohistochemical method optimized for clinical use, desmoid can be regarded as a c-Kit-negative tumor. PMID- 12645335 TI - Steroid hormone regulation and prognostic value of the human kallikrein gene 14 in ovarian cancer. AB - To study KLK14 gene expression in endocrine-related cancers, we studied its hormonal regulation in breast and ovarian cancer cell lines. Our kinetic and blocking experiments suggest that this up-regulation is mediated through the androgen receptor. We then studied the expression of KLK14 by quantitative reverse transcriptase-polymerase chain reaction in 155 consecutive ovarian tumors and correlated these findings with clinicopathologic parameters, response to chemotherapy, and survival. A stepwise reduction was observed in the levels of KLK14 messenger RNA in normal, benign, and cancerous tissues (P < .001). Expression levels were significantly higher in patients with early stage disease and optimal debulking and in patients who responded to chemotherapy. Kaplan-Meier survival curves demonstrated longer progression-free and overall survival in patients with KLK14-positive tumors than in patients with KLK14-negative tumors (P < .001). When all other prognostic variables were controlled in the multivariate analysis, KLK14 retained its prognostic significance (progression free and overall survival, respectively, hazard ratios, 0.43 and 0.53; P = .027 and .014). A weak negative correlation was found between KLK14 expression and serum CA-125. KLK14 is a new, independent, and favorable prognostic marker for ovarian cancer. PMID- 12645336 TI - Accuracy and reproducibility of telecytology diagnosis of cervical smears. A tool for quality assurance programs. AB - We randomly selected 50 cervical smears (benign, 14; atypical squamous cells of undetermined significance [ASCUS], 5; low-grade squamous intraepithelial lesion [LSIL], 10; high-grade squamous intraepithelial lesion (HSIL), 12; squamous cell carcinoma, 6; adenocarcinoma, 3) and captured 1,181 digital images (518 MB) at a maximum resolution of 1,600 x 1,200 pixels and transmitted them by e-mail. Diagnosis of glass slides and digital images was done independently in a double blind manner by 3 pathologists and 3 cytotechnologists, commencing with the diagnosis of digital images followed by diagnosis of glass slides 3 months later. The procedure was repeated after 3 months. Diagnoses were recorded as benign, ASCUS or atypical glandular cells of undetermined significance, LSIL, HSIL, squamous cell carcinoma or adenocarcinoma, and "inadequate for diagnosis." Diagnostic accuracy and interobserver reproducibility were analyzed using an intraclass correlation coefficient (ICC), which revealed good interobserver agreement for the first (0.72) and second (0.64) glass slide diagnoses and the first (0.72) and second (0.60) digital image diagnoses. The kappa values for intraobserver variation between first and second glass slide diagnoses and first and second digital image diagnoses showed moderate to excellent agreement. Digital images are suitable substitutes for glass slides; telecytology can be used as an alternative method for the cytologic diagnosis of cervical smears, particularly in quality assurance programs. PMID- 12645337 TI - Immunoreactivity of Hep Par 1 in hepatic and extrahepatic tumors and its correlation with albumin in situ hybridization in hepatocellular carcinoma. AB - We evaluated the expression of Hep Par 1 (hepatocyte paraffin 1 monoclonal antibody) in 42 hepatocellular carcinomas (HCCs), 25 cholangiocarcinomas, 18 tumors metastatic to the liver, and 87 primary extrahepatic tumors. Albumin in situ hybridization (ISH) was performed in the HCC cases. Of 42 cases of HCC, 39 (93%) were positive for Hep Par 1. All cases of cholangiocarcinoma, renal cell carcinoma, adrenocortical carcinoma, and islet cell tumors were negative; 1 case each of primary urinary bladder (n = 10) and pancreatic (n = 10) adenocarcinoma and 3 of 11 cases of primary pulmonary adenocarcinoma showed focal positivity; 7 of 10 gastric and 6 of 8 esophageal adenocarcinomas were strongly positive. Albumin ISH was positive in 39 (93%) HCC cases. All cases of HCC were positive for Hep Par 1 or albumin ISH. Hep Par 1 immunoreactivity has high sensitivity in the diagnosis of HCC. Strong positive staining also occurs in gastroesophageal adenocarcinomas. Cholangiocarcinoma and carcinomas from most other sites are negative for Hep Par 1. Hep Par 1 immunoreactivity shows high correlation with albumin ISH; their combined use for diagnosis of HCC had a sensitivity of 100% in this population. PMID- 12645338 TI - Interobserver variability in assessing adequacy of the squamous component in conventional cervicovaginal smears. AB - We compared the interobserver reproducibility of estimating the adequacy of the squamous component of conventional Papanicolaou (Pap) smears using traditional and newly proposed criteria. Forty conventional Pap smears with varying degrees of squamous cellularity were reviewed by 13 observers who evaluated adequacy (satisfactory vs unsatisfactory) based on the traditional criterion of estimating 10% slide coverage. After being introduced to the new criterion and the reference images, the observers reevaluated adequacy on the same set of smears, using the new criterion and the reference images. With the original criterion of 10% slide coverage, 15 smears had a unanimous designation; the overall kappa value was 0.49 (P < .001). With the newly proposed adequacy criterion and reference images, 17 smears had a unanimous designation; the overall kappa value was 0.60 (P < .001). The difference in the kappa correlation coefficients was statistically significant (P = .007). While traditional and newly proposed criteria resulted in fair interobserver agreement, it seemed that the newly proposed criterion, along with the use of reference images, for evaluating adequacy of the squamous component of conventional Pap smears results in better interobserver reproducibility. PMID- 12645339 TI - CD105 expression is a marker of high metastatic risk and poor outcome in breast carcinomas. Correlations between immunohistochemical analysis and long-term follow-up in a series of 929 patients. AB - CD105 (endoglin) is expressed significantly in activated endothelial cells in culture and in tumor microvessels. Quantification of CD105 immunocytochemical expression that may be clinically relevant has not been accurately evaluated. We studied CD105 expression on frozen tissue sections by using immunohistochemical assays in a series of 929 patients and correlated the findings with long-term follow-up (median, 11.3 years). Univariate (Kaplan-Meier) analysis showed that the number of CD105+ microvessels (cutoff, 15 vessels) correlated significantly with poor overall survival among all patients (P = .001). This correlation was less significant in node-negative patients (P = .035). Marked CD105 expression also correlated with a high risk for metastasis among all patients (P = .006) and among node-negative patients (P = .001). Multivariate analysis (Cox model) identified CD105 immunodetection as an independent prognostic indicator. Our results suggest that immunohistochemical expression of CD105 has practical clinical relevance for identifying node-negative patients with a poor prognosis. Moreover, immunodetection of CD105 also may be considered a potential tool for selecting patients who could benefit from specific antiangiogenic therapy, using anti-CD105 conjugates. PMID- 12645340 TI - Comparison of fluorescence and chromogenic in situ hybridization for detection of HER-2/neu oncogene in breast cancer. AB - Determination of HER-2/neu oncogene amplification has become clinically important in the present management of breast cancer and may have important applications in other areas of clinical oncology and scientific research. In situ hybridization is an extremely accurate and sensitive technique for assessing amplification of HER-2/neu. A new method using a chromogen-labeled probe offers numerous advantages, including the ability to view the morphologic features of the cells of interest using a light microscope, which can be found in every laboratory. We used both techniques to assay 31 cases of infiltrating breast carcinoma; assays were performed in laboratories at 2 institutions. Identical results for both methods were found in 26 cases (10 amplified, 16 nonamplified). One case was misinterpreted as overexpressed by chromogenic in situ hybridization (CISH) because of background precipitate. In 4 cases, CISH suggested low-level amplification. Three of these cases subsequently were found to have chromosome 17 polysomy. In the remaining case, the initial section chosen was suboptimal, showing weak signals by both methods. If a probe for chromosome 17 (now available for CISH) is used in cases of questionable HER-2/neu amplification, CISH seems to be as accurate and more practical than FISH. PMID- 12645341 TI - How many cases need to be reviewed to compare performance in surgical pathology? AB - Recent studies have shown increased interest in measuring error rates in surgical pathology. We sought to determine how many surgical pathology cases need to be reviewed to show a significant difference from published error rates for review of routine or biopsy cases. Results of 4 series with this type of diagnostic material involving a total of 11,683 cases were reviewed to determine the range of published false-negative, false-positive, typing error, threshold error, and clinically significant error rates. Error rates ranged from 0.00% to 2.36%; clinically significant error rates ranged from 0.34% to 1.19%. Assuming a power of 0.80 and a 1-sided alpha of 0.05, the number of cases needed to be reviewed to show that a laboratory with either twice or one half the published error rate was significantly different from the range of published error rates varied from 3.30 to 50, 158. For clinically significant errors, the number of cases varied from 665 to 5,886. Because the published error rates are low, a relatively large number of cases need to be reviewed and a relatively great difference in error rate needs to exist to show a significant difference in performance in surgical pathology. PMID- 12645342 TI - Expression patterns of protein kinase C isoenzymes are characteristically modulated in chronic pancreatitis and pancreatic cancer. AB - We immunohistochemically identified protein kinase C (PKC) isoenzymes and the receptor for activated C-kinase (RACK-1) in normal, chronically inflamed, and malignant pancreas specimens. Expression patterns were specific and consistent for each microanatomic structure. In chronic pancreatitis, the expression patterns by epithelial cells were indistinguishable from those in normal pancreas. In the stroma, there was a gain of PKC-delta (P < .05) and loss of PKC mu (P < .0001). Expression in pancreatic duct carcinomas, compared with control normal minor ductular epithelial cells, revealed relative loss of PKC-epsilon (P < .0001), PKC-iota (P = .005), and PKC-theta (P < .0001) but no gain in any isoenzyme. Compared with control normal major duct epithelial cells, the principal differences were a relative loss in PKC-gamma (P < .05) and a relative gain in PKC-beta (P < .05), PKC-iota (P < .05), and PKC-zeta (P < .005). The stroma adjacent to ductal carcinomas was characterized by prominent expression of PKC-mu and a gain in PKC-delta (P < .0001) and PKC-zeta (P > .005). Ampullary carcinomas revealed a relative gain of PKC-iota (P < .05) and RACK-1 (P < .05). In the adjacent stroma was enhanced expression of PKC-delta (P < .005) and PKC gamma (P < .001) and loss of PKC-mu (P < .05). Specific changes in isoenzyme expression in stroma of chronic pancreatitis and in epithelial cells and stroma of ductal and ampullary pancreatic adenocarcinomas reflect specific modulation of intracellular signaling pathways that control critical homeostatic mechanisms. PMID- 12645343 TI - Helicobacter pylori. Pathology and diagnostic strategies. AB - Helicobacter pylori represents one of the most common and medically prominent infections worldwide. Infection with this microaerobic, gram-negative bacterium has been established as an etiologic factor in the development of peptic ulcer disease. In addition, H pylori infection has been associated firmly with the development of gastric neoplasia, including gastric adenocarcinomas and gastric mucosa-associated lymphoid tissue lymphomas. Effective antimicrobial treatment depends on sensitive and accurate diagnostic approaches. This review article discusses invasive and noninvasive strategies for diagnosis of H pylori infection. Invasive methods requiring endoscopic evaluation include bacteriologic culture and susceptibility testing, histopathologic studies, molecular diagnostics, and rapid urease testing. Noninvasive approaches include fecal antigen detection, serologic testing, and urea breath testing. PMID- 12645344 TI - Bone marrow histopathologic and molecular staging in epidermotropic T-cell lymphomas. AB - This study was undertaken to determine the prognostic value of bone marrow histopathologic and molecular analyses in 53 patients with mycosis fungoides and 7 with Sezary syndrome. Bone marrow was involved in only 1 patient with Sezary syndrome, clinical stage IVA, before bone marrow biopsy. An ambiguous T-cell infiltrate was observed in 8 patients but was not associated with disease progression. The bone marrow specimen was normal in 51 patients. Monoclonality was detected in the skin specimen in 44 cases; an identical T-cell clone in the blood specimen was found in 21 of them and, in 16 of the 21 patients, in bone marrow specimens without histologic correlation. Multivariate analysis confirmed that clinical stage and detection by polymerase chain reaction of an identical T cell clone in skin and blood specimens had an independent prognostic value. No further prognostic value was observed for the presence of a T-cell clone in bone marrow specimens. Our data do not support the need for bone marrow examination in patients with mycosis fungoides/Sezary syndrome. PMID- 12645345 TI - CXCR4/CD184 immunoreactivity in T-cell non-Hodgkin lymphomas with an overall Th1- Th2+ immunophenotype. AB - We performed an immunohistochemical analysis of CXCR4/CD184 expression in frozen and paraffin-embedded sections of human peripheral T-cell lymphomas that exhibit a composite Th1 T-cell-like or Th2 T-cell-like immunophenotype, based on expression of Th1-associated markers (CXCR3, OX40/CD134, and CD69) and Th2 associated markers (CD30 and CCR4). In 66 cases examined, CXCR4/CD184 expression correlated significantly with immunoreactivity for other markers of Th2 differentiation (P < .0001). Anaplastic large cell lymphoma, which typically expresses markers of Th2 differentiation, was immunoreactive for CXCR4/CD184 in 22 (88%) of 25 cases. Tumors previously identified as exhibiting a composite Th1 like immunophenotype, which include angioimmunoblastic lymphoma, lymphoepithelioid lymphoma, and other peripheral T-cell lymphomas now termed unspecified, were positive for CXCR4/CD184 in 7 (17%) of 41 cases. These results are consistent with previous findings that a subset of peripheral T-cell lymphomas can be divided into Th1-like and Th2-like categories based on immunoreactivity with a limited set of markers. Our findings also suggest that CXCR4/CD184, which is expressed by a number of malignant neoplasms and may have a role in tumor metastasis, may have a similar function in CXCR4+ T-cell non Hodgkin lymphomas. PMID- 12645348 TI - Diagnostic certainty is sometimes certainly an error. AB - We evaluated a low-density lipoprotein (LDL) subfraction separation method using polyacrylamide tube gel electrophoresis (PTGE) and compared it with the reference method, polyacrylamide gradient gel electrophoresis (PGGE-REF). Excellent intra assay and interassay coefficients of variation were obtained (<4%) for PTGE. For 102 subjects, LDL subclasses correlated most significantly with triglyceride (TG) level, high-density lipoprotein (HDL) cholesterol level, total cholesterol/HDL cholesterol ratio, and non-HDL cholesterol level (P < .05). The distribution of large LDL (76%) was predominant for subjects with low TG levels (< 150 mg/dL [1.69 mmol/L]), while distribution of small LDL (79%) was predominant for subjects with high TG levels (>200 mg/dL [2.26 mmol/L]). Excellent agreement between the methods was observed (weighted kappa = 0.78). Of 51 samples classified as small, dense LDL by PGGE-REF, none were misclassified as lage LDL and 4 as intermediate LDL by PTGE (92% concordance); of 44 samples classified as small and 7 as intermediate by PTGE (77% concordance). The PTGE method is precise and compares favorably with PGGE-REF. It has the advantage of being simple, less expensive, and more suitable for use in the clinical laboratory. PMID- 12645347 TI - A computer-based intervention for improving the appropriateness of antiepileptic drug level monitoring. AB - We designed and implemented 2 automated, computerized screens for use at the time of antiepileptic drug (AED) test order entry to improve appropriateness by reminding physicians when a potentially redundant test was ordered and providing common indications for monitoring and pharmacokinetics of the specific AED. All computerized orders for inpatient serum AED levels during two 3-month periods were included in the study. During the 3-month period after implementation of the automated intervention, 13% of all AED tests ordered were canceled following computerized reminders. For orders appearing redundant, the cancellation rate was 27%. For nonredundant orders, 4% were canceled when information on specific AED monitoring and pharmacokinetics was provided. The cancellation rate was sustained after 4 years. There has been a 19.5% decrease in total AED testing volume since implementation of this intervention, despite a 19.3% increase in overall chemistry test volume. Inappropriateness owing to repeated testing before pharmacologic steady state was reached decreased from 54% of all AED orders to 14.6%. A simple, automated, activity-based intervention targeting a specific test ordering behavior effectively reduced inappropriate laboratory testing. The sustained benefit supports the idea that computerized interventions may durably affect physician behavior. Computerized delivery of such evidence-based boundary guidelines can help narrow the gap between evidence and practice. PMID- 12645349 TI - [Delirium is certainly not an unavoidable complication of pain control in the terminal phase of life]. AB - In three terminal patients, a man aged 19 years who suffered from progressive osteosarcoma, a man aged 71 years with a small-cell pulmonary carcinoma, and a 68 year-old woman with cerebral metastases from a mammary carcinoma, delirium developed due to increased dosage of opioids for seemingly intractable pain (the first two patients) and dexamethasone (third patient). The delirium subsided after opioid rotation, administration of drugs for neuropathic pain, and treatment with an antipsychotic, respectively. This enhanced the patients' quality of terminal life and quality of dying. In terminal patients, analgesics induced delirium must be considered, diagnosed and treated without delay. PMID- 12645350 TI - [Health Council of the Netherlands' report on contraception in intellectually disabled people]. AB - Recently the Health Council of the Netherlands published a comprehensive report which incorporates juridical, ethical, medical and social aspects directing the judgement and decision-making regarding contraception in people with an intellectual disability. Although professional responsibility is in the hands of medical practitioners, medical guidelines to manage consent issues and to indicate contraception in this population are missing. There are no ethical and legal grounds for enforcing contraception and it comes down to professional accountability in trying to prevent irresponsible parenthood by a convincing dialogue. People with intellectual disabilities are intellectually, physically, socially and economically prone to be vulnerable parents, who need assistance in making deliberate choices to have children and in raising them. There is an urgent need for advanced sexual education and a comprehensive social discussion on contraception for this population. Physicians for intellectually disabled people embrace the committee's recommendations to stimulate public debate, enhance research, develop guidelines and build professional networks to call on. PMID- 12645351 TI - [Early diagnosis of rheumatoid arthritis with a test based upon a specific antigen: cyclic citrullinated peptide]. AB - In patients with rheumatoid arthritis (RA), joint erosions occur at a very early stage of the disease before clinical symptoms can be detected. Early treatment with currently available antirheumatic drugs may stop or delay the development of such erosions. A simple and specific diagnostic test is needed for treatment to be initiated at an early stage. The specificity of the routinely used rheumatoid factor (RF) test is too low for that purpose. A novel autoantibody, directed to citrullinated antigens in the synovium, seems to provide a new starting point. These citrullinated autoantigens (e.g. fibrin) are specifically present in inflamed synovia and the antibodies for these are locally produced. The autoantibodies can be detected in the blood of the patients with RA years before the first clinical signs are manifest, and high titres appear to correlate strongly with erosive disease. The test for cyclic citrullinated peptide, which has recently become available, has a specificity of 98-99% and a sensitivity of 75-80%. PMID- 12645352 TI - [Diagnosis of vegetative state as a basis for medical treatment on the borderline between life and death]. AB - The term 'vegetative state' is most appropriate for the state which develops when patients open their eyes after a comatose phase, without regaining consciousness. The definition and the diagnostic criteria from the Multi Society Task Force on Persistent Vegetative State are usable for the clinical practice in the Netherlands. The vegetative state must be differentiated from coma, locked-in syndrome and minimally conscious state. A systematic, multidisciplinary approach under the direction of a physician is key to diagnosing vegetative state. To this end, a clinical assessment is recommended with reassessment and verification of the diagnosis at appropriate moments. Careful observation remains the fundamental to the diagnosis. PMID- 12645353 TI - [Submental intubation: surgical and anesthesiological aspects]. AB - In patients with cranio-maxillofacial trauma and in corrections of cranio-facial anomalies submental intubation may be used during surgery and postoperatively. In the period June 2001-May 2002 submental intubation was performed on five patients with cranio-maxillofacial trauma and on two patients with cranio-facial anomalies (Dubowitz syndrome (LeFort II osteotomy and distraction) in one patient and cheilognathopalatoschisis (Le Fort I osteotomy and distraction) in the other). In accordance with the literature the submental intubation technique was a useful, fast and safe technique which provides a secure airway during surgery. No postoperative intubation-related complications were seen in the group of patients. It provides the surgeon with an excellent view of the operation field and permits optimal intra-operative control of the dental occlusion. Combined procedures by the surgeon and the anaesthetist must be planned beforehand. PMID- 12645354 TI - [Diagnostic image (124) A man with hemiballism. Hemiballism caused by an contralateral infarct in the mesencephalon region of the nucleus subthalamus]. AB - A 65-year-old man presented with hemiballism due to an infarct in the contralateral subthalamic region. PMID- 12645355 TI - [Clinical treatment of equestrian injuries in Groningen, 1990-1998: serious long term effects]. AB - OBJECTIVE: To determine whether equestrian injuries result in long-term residual symptoms and to establish the risk factors for these outcomes. DESIGN: Retrospective, descriptive. METHOD: A total of 88 patients older than 17 years of age who had been admitted to the Department of Surgery, University Hospital Groningen during the period 1990-1998 because of an equestrian injury were included in the study. After an average period of five years an inventory was made using the Sickness Impact Profile 68 (SIP68), if absenteeism from work or sport and disabilities or handicaps were experienced. Following this the risk factors for the occurrence of long-term effects were investigated. RESULTS: The response was 65/88 (74%). Equestrian injuries often resulted in long-term effects: 35% of the patients had a SIP68 score > 0.11% were permanently unfit for work, 25% no longer participated in sport as an effect of the injury and 43% still experienced some form of disability or handicap; on average this was all 5 years after the injury. The type of injury (notably intracranial, fracture), body region (notably trunk), and the injury severity score (ISS) were significantly correlated with the occurrence of long-term effects. CONCLUSION: Equestrian injuries which required clinical treatment were serious in nature and often resulted in disabilities and handicaps. PMID- 12645357 TI - [How to treat a patient with indications for an infectious viral hemorrhagic fever]. PMID- 12645356 TI - [Sport drinks: not a suitable rehydration solution for children]. AB - A two-months-old male infant with gastroenteritis had been given a sports drink (Aquarius) by the general practitioner as a treatment for dehydration and was referred because of ongoing vomiting and weight loss. The child recovered after reinstitution of normal bottle-feeding and administration of a regular oral rehydration solution (ORS). The sports drink was analysed and found to contain less salt than ORS. As a result, the salt:sugar ratio was not optimal. Sports drinks are therefore contraindicated in the treatment of dehydration in children. PMID- 12645358 TI - [Nutrition and health--fish fatty acids against fatal coronary heart disease]. PMID- 12645359 TI - [Contacts outside of office hours: complaints presented to the general practitioner and to the emergency department]. PMID- 12645360 TI - [Favorable treatment with dexamethasone in adults with acute bacterial meningitis: a randomized, placebo controlled trial]. PMID- 12645361 TI - Breastfeeding technique in prevention of nipple sore--a study. PMID- 12645362 TI - Computers in nursing. PMID- 12645364 TI - World Breastfeeding Week: August 1-7, 2002. 'Healthy mothers and healthy babies'. PMID- 12645363 TI - Wound dressing practices--a study. PMID- 12645365 TI - Loss prevention case of the month. Good care first, then good documentation. PMID- 12645366 TI - HIV/AIDS medication adherence factors: inner-city clinic patient's self-reports. AB - OBJECTIVE: To document common facilitators of, and barriers to, HIV/AIDS medication regimen adherence and to identify facilitators and barriers significantly correlated to attainment of higher vs. lower adherence. DESIGN: Data were collected using semi-structured interviews. SETTING: An outpatient clinic serving as the regional treatment center for HIV disease. PATIENTS: A purposive sample of 57 clinic patients was enrolled and completed the study. MAIN OUTCOME MEASURES: Self-reported adherence was measured using a visual analog scale and specified as higher or lower adherence at the 80% level, the approximate median. RESULTS: Many barriers to adherence were more prevalent in the lower adherence group, including complexity of the medication regimen and experiencing side effects. Lower adherence patients also had more problems with privacy and interference with social life and work or school. Patients in the higher adherence group were more concerned with forgetting as an adherence barrier. In general, social support, motivation to avoid AIDS, perceiving the health care practitioner as a facilitator, knowledge of medications, and keeping schedules were identified as factors enhancing adherence. CONCLUSIONS: The high prevalence of many barriers shows that the health care provider has an important role in patient education and support to accomplish increased adherence. Fear and avoidance behaviors in the lower adherence group contrasted with the increased concern with forgetting in the higher adherence group. Communicating treatment advances with patients may help more patients to address many barriers to higher adherence. PMID- 12645367 TI - Heart failure: do we deliver quality care? PMID- 12645368 TI - Emergency defibrillation: a cornerstone of care for Tennessee Emergency Medical Services. PMID- 12645369 TI - OxyContin: maintaining availability and efficacy while preventing diversion and abuse. PMID- 12645370 TI - [Urinary obstruction]. AB - Obstructive uropathy is a result of the particular tubular configuration of the urinary tract and involves virtually all its organs. This review analyses upper urinary tract obstructions as well as a number of lower urinary tract conditions whenever the circumstances of the urological disease progress so advises it. A comprehensive exposition is made of the different types of obstruction, the basic pathophysiological principles and the respective anatomical transposition. The pathophysiological features characterising intrauterine obstructions, obstructions occurring during pregnancy and in the elderly are also highlighted. Finally an updated analysis is made of the diagnostic contribution made by all techniques susceptible to be used when dealing with obstructive uropathies highlighting the prognostic relevance of early diagnosis and treatment. The hope placed in the use of modern techniques to establish a differential diagnosis of chronic obstructive uropathy is also emphasised. PMID- 12645371 TI - [Present and future of kidney transplantation]. AB - Renal transplant is the treatment of choice for the patient with end stage renal disease. Spain is the country with the highest donation rate (33 ppm). However, at present this figure is stabilized. The development of non-beating heart programmes, living-donor nephrectomy (specially laparoscopic nephrectomy) programmes, and may be xenotransplantation in a non-immediate future could increase the transplantation activity. The knowledge of preservation mechanisms, specially with the use of perfusion machines allows to rescue for transplantation kidneys with a long warm-ischemia time. Furthermore, these machines are useful for analyzing viability markers. The new immunosuppressive drugs: Tacrolimus, Mycophenolate-Mophetil, Rapamycin and monoclonal antibodies against alpha chain of the interleukine-2 receptor (Basoliximab and Dazcizumab) have reduced the incidence of acute rejection in the immediate renal transplant period. However, its effect in the long-term follow-up period is still a matter of controversy. The incidence of tumour in the renal transplant recipient is increased, specially those of lymphoma, skin cancer and Kaposi sarcoma. Periodical exams for detecting the development of tumours are mandatory in this population. Finally, xenotransplantation is an attractive alternative, although immunological, infective and ethical barriers should previously be resolved. PMID- 12645372 TI - [Progress in the treatment of germ cell tumor of the testis]. AB - Breakthroughs seen over recent years in the management of testicle germ cell tumours have been very significant. The prognostic influence of such advances has proven to be crucial and translates the progressive opening of oncology to new fields and technologies. In his review, the author articulates a potential target in the management of these germ cells tumours to further increase survival rates. The therapeutic challenges include the consideration of prognostic factors, decreased toxicity of established therapies, addition of new molecules to first line therapy and the recent application of molecular biology techniques to both diagnostic and therapeutic aspects. PMID- 12645373 TI - [Neoplastic pathology of the urothelium]. AB - Bladder cancer is one of the most common diseases treated by urologists. In this article, we will try to review some of the controversies and all the available data which come from the systematic review and meta-analysis. PMID- 12645374 TI - [Aging and urology: is impotence the first symptom of aging?]. AB - The clinical expression of ageing involves several organs with variable degrees of precociousness or intensity based on the apparatus or system. An early event in the ageing process may be the occurrence of asymptomatic, and difficult to recognise, periurethral hyperplastic nodules. On the contrary, such an obvious symptom and clinical entity as erectile dysfunction may become a highly relevant herald of ageing. And this will be more so if it appears in coincidence with widespread cardiovascular disease, specially if manifest through hypertension and ischaemic heart disease. The action of the urologist faced with a patient who asks for help for his erectile dysfunction cannot be limited to the evaluation of a simple organ disease. PMID- 12645375 TI - [Urinary fistula: update]. AB - The urinary fistulas are not a frequent problem in our urological rutinary work, being the vesicovaginal ones the most common. However, it will be necessary to know them to be able to diagnose and treat them in the best way. In this article we classificate the different urinary fistulas (uro-gynecologicals, uro vasculars, uro-cutaneous and entero-urinarys), explaining the possible etiologies and the correct management. PMID- 12645376 TI - [Bilateral synchronous renal cell carcinoma. Report of a case]. AB - The radical nephrectomy is considered to be the optional treatment for renal carcinoma. In the last years, the good results with partial resection shows conservadora surgery the most suitable treatment for well-localized bilateral renal tumours. In this cases dialysis is avoided, and the subsequent risk of immunological deficit and the higher possibility of metastasic illness are minimized. Nowadays, however, there seems to be some controversy over conservative surgery, concerning applications and technique of surgery to practice (partial resection vs. enucleation). We have a case of bilateral synchronic renal cell carcinoma where radical surgery on the left kidney was put into practice, as it presented a great tumour with multiples points corticals confirmed by PAAF and conservative surgery on the right kidney with enucleation of localized mass in the lower renal. The patient held normal renal function with a good quality of life for five years, finding out then, in one of the radiological tests, carcinoma relapse in the only kidney, that forced to radical surgery and to his inclusion in dialysis programme. PMID- 12645377 TI - [Infertility caused by AZF microdeletions. A new case of azoospermia]. AB - We report report a case of a man with azoospermia who was studied because of infertility. Between the causes of azoospermia and severe oligospermia, in male patients with infertility evaluations, small deletions in regions of the Y chromosome must be considered, as in our case. Regions for spermatogenesis in the Yq chromosome can be evidenced using PCR analysis. Men with azoospermia are at high risk to have small deletions in that regions. Yq chromosome deletions could be transmitted when ICSI (intracytoplasmic sperm injection) is applied. Thereafter, study of microdeletions would be included in severe infertile men. PMID- 12645378 TI - [Traumatic testicular dislocation. Report of a case]. AB - Testicular dislocation after blunt scrotal trauma is a rare event. This injure must be recognized and treated promptly to maximize the likelihood of testicular salvage. We describe a new case of unnoticed traumatic testicular dislocation with secondary testicular lost. We discuss the chain of events leading toward the correct diagnosis and treatment based on a review of published reports. PMID- 12645379 TI - [Acute diffuse bacterial nephritis in horse shoe kidney]. AB - Horseshoe kidney is a congenital malformation which predisposes the patient to severe urinary tract infections, due to pelvic ectasia which is inherent in the malrotation of the renal units. Acute bacterial nephritis is an evolutionary stage of the acute renal infection situated between acute pyelonephritis and renal abscess. It is only possible to carry out a differential diagnosis of these two bodies through the application of some imaging technique, as it is impossible to differentiate them from a clinical point of view. A CT contrast scan is the most sensitive and specific means of diagnosing acute bacterial nephritis, and is considered to be the most appropriate test method. The CT scan usually reveals circular or wedge-shaped cortical areas which become striated after contrast administration. Treatment is based on broad-spectrum antibiotherapy, which must be continued for a period of 3 to 6 weeks. We present a case of diffuse acute bacterial nephritis, occurring in a paediatric patient with a horseshoe kidney, which was successfully treated by means of a prolonged course of antibiotics. PMID- 12645381 TI - [Syphilitic penile pseudotumor. Report of a case]. AB - The syphilis is a venereal disease of sexual transmission that incidence is increasing in several groups populations. We report a case of a patient with secondary syphilis who consulted for an atypical solid mass to penile. The clinical evolution, histological examination and analytical tests confirmed the diagnosis of genital syphilis having a satisfactory response to specific antibiotic treatment. The epidemiological, clinical, diagnostic aspects and therapeutic of the genital ulcer disease are discussed. PMID- 12645382 TI - [Recommendations of the European Society of Urogenital Radiology on the use of contrast media and the opportunity to use paramagnetic contrast in patients allergic to iodinated contrast media treated with endourology]. PMID- 12645383 TI - [Common biostatistical errors in clinical studies]. AB - Roughly half to two third of all published biomedical studies that use statistical methods contain unacceptable errors. The present article points at common errors that may be avoided without requiring profound statistical knowledge. These errors mainly concern the minimal number of patients and sample size (statistical power), agreement between aim and conclusion, distribution of data as well as description of location and variability of data. An analysis of 150 papers in the New England Journal of Medicine and in Circulation demonstrates that these errors can also commonly be found in respected journals after statistical peer review. Editors of biomedical journals could reduce the problem by means of statistical guidelines. PMID- 12645380 TI - [Small cell carcinoma of the bladder. Report of a new case and review of the literature]. AB - Small cell carcinoma of the bladder is a rare entity occurring in fewer than 1% of all the primary bladder tumors. Its clinic and epidemiologyc characteristics are similar to the transitional cell carcinoma. But, its more aggressive with a high tendency to infiltrate the muscular layer and to metastasize. Thus, therapeutic approach used by the pulmonary form has been applied to the bladder carcinoma. The best results has been achieved using systemic chemotherapy, not only in survival rates but in preserving the bladder integrity, playing the radical cystectomy a rescue role in cases of recurrence or persistence of the tumor. We report a new case of small cell carcinoma of the bladder and review the literature with special attention to the different therapeutic approaches. PMID- 12645384 TI - [Combined functional and morphological imaging with PET/CT systems]. PMID- 12645385 TI - [Health and illness of Kosovo migrants. Indications for medical consultation]. PMID- 12645386 TI - [Muscle weakness]. AB - A 31-year old cook suffered from episodes with muscular weakness since 3 weeks. These episodes resolved, at times, spontaneously and completely after a few hours. The weakness affected mainly the muscles of trunk and proximal extremities. Investigation of the patient revealed flaccid tetraparesis with diminished tendon reflexes, however, cranial muscles were not affected. Mental function and sensibility were intact. These findings were consistent with hypopotassemic periodic paralysis rather than a neurological or psychiatric disease. Serum potassium was determined as 1.8 mmol/l. Moreover, the patient suffered from atrial fibrillation. Grave's disease could be identified as the underlying cause of hypopotasemic periodic paralysis and atrial fibrillation. Further manifestations of primary hyperthyroidism were nervousness, diarrhea and weight loss of 5 kg observed during the previous months. The symptoms resolved under treatment with potassium, betablocker and thyrostatic agents. The patient was treated with warfarin because of atrial fibrillation and finally underwent electroconversion. PMID- 12645387 TI - [A 45-year-old woman patient with thigh pain]. PMID- 12645388 TI - [A 30-year-old woman patient with reno-parenchymatous arterial hypertension]. PMID- 12645389 TI - [Therapeutic approach to complex regional pain syndrome]. PMID- 12645390 TI - [Cloning from the relational viewpoint]. PMID- 12645391 TI - [Pharmacological sympathetic block in complex regional pain syndrome]. AB - PURPOSE: The Complex Regional Pain Syndrome (CRPS) is a chronic pain state provoked by lesions of the soft tissues or of the bony tissues (type CRPS-I or reflex sympathetic dystrophy-RSD) or by lesions of the nerves (type CRPS-II or causalgia) with vegetative alterations (perspiration, vasomotory alterations) and trophic alterations (bony cutaneous atrophy, alopecia, articular contractures). The pharmacological block of the sympathetic nerves through a peripheral vein is inserted in the multidisciplinary approach that characterizes the therapy of this syndrome. MATERIALS AND METHODS: A retrospective survey was carried out on a group of 185 patients affected by RDS/CRPS with block of the sympathetic nerves through a peripheral vein with guanethidine. Superior limb: Inflation of the tourniquet till disappearance of the radial wrist. Cannulation of a peripheral vein with Butterfly needle n. 23. Guanethidine 10 mg, lidocaine 20 mg, sodic heparin 500 u.i, NaCl 0.9% 20 ml. Injection in 5 minutes. Permanence of the pneumatic tourniquet inflated above systolic blood pressure for 15 minutes. Deflation slowly. Inferior limb: Inflation of the tourniquet till disappearance of the pedidium wrist. Cannulation of a peripheral vein with Butterfly needle n. 23. Guanethidine 20 mg, lidocaine 40 mg, sodic heparin 1000 u.i, NaCl 0.9% 40 ml. Injection in 5 minutes. Permanence of the pneumatic tourniquet inflated above systolic blood pressure for 15 minutes. Deflation slowly. RESULTS: The first stage (hyperemic) showed the highest incidence of remissions: (83, 33%). Even in the second stage (dystrophic) the answer to the therapy has been fundamentally positive: (53, 68%). In the third stage (atrophic) the results have been more modest: (8, 33%). CONCLUSIONS: The block of sympathetic system with guanethidine is still an important method in the therapy of the CRPS; in fact it is surely less invading than the blocks of the stellate ganglion or of the lumbar sympathetic. PMID- 12645392 TI - [Changes in tumor markers CEA, Ca 19-9 and Ca 125 in monitoring of response to chemotherapy in elderly patients with advanced gastric cancer]. AB - PURPOSE: We have evaluated whether tumor markers can be used to assess response to systemic chemotherapy. We analyzed the relationship between the response to chemotherapy based on serial imaging and on change in serum tumor marker level of CEA Ca 19-9 and Ca 125. PATIENTS AND METHODS: The levels of serum CEA, Ca 19.9 and Ca 125 were measured before and after 6 and 12 week systemic chemotherapy in twenty-five elderly patients age 68-80 (median age 71) with advanced gastric cancer. We evaluated the relationship between the change in serum tumor markers level and response assessment by imaging studies throughout the treatment course. RESULTS: There was a significant correlation between the assessment of response by tumor markers and by imaging studies. The survival time of responders assessed by tumor markers was significantly longer than that of non-responders. CONCLUSIONS: The serum levels tumor markers may be used as a mean of monitoring treatment in elderly patients, when in a imaging study it is difficult, to assess response to chemotherapy and in predicting the prognosis of patients with advanced gastric cancer. PMID- 12645394 TI - [Ectropion of the uterine cervix in adolescence. Study of a topical drug as an alternative to surgical treatment]. AB - The Authors, studied the ectropion of the uterine cervix in the teen-agers (young ladies) and the effect of the desossiribonucleic acid therapy in topic way by 5 mg/die for 20 days in 80 patients with an ectropion of the I degree and the II degree portion of the uterine cervix. The patients were separated in two groups. 40 patients treated by desossiribonucleic acid therapy, 40 patients didn't take any medicament. In conclusion the results show that the desossiribonucleic therapy could be an alternative way for the treatment of the ectropion of the uterine cervix. PMID- 12645395 TI - [Ethical aspects in medical education and updating of medicine]. PMID- 12645393 TI - [Effectiveness and tolerability of selegiline in the treatment of pathological cerebral involutions]. AB - A dysfunction of dopaminergic systems with reduction of active dopamine, is well known in cerebral involution. Twenty patients with mild-moderate pathological cerebral involution of atrophic and/or vascular origin, were treated with Selegiline (L-deprenyl), a monoamino-oxidase B inhibitor (10 mg/die for six months). Compared with a control group, Selegiline treated patients showed a statistically significant improvement in cognitive and behaviour capacities. At the end of investigation, "Mini Mental State" showed an improvement of 26.5% in Selegiline group and of 3.7% in control group (P < 0.01). "Echelle Clinique d'Aptitudes Intellectuelles" showed an improvement of 29.4% and of 10.8% respectively (P < 0.01). Selegiline treatment has shortened significantly the reaction times and has improved mnesic capacities. No side effects were observed during the study. PMID- 12645396 TI - [The nurse's role in the identification and management of alcohol abuser in the Emergency Department]. AB - The emergency department (ED) nurse plays a major role both in recognizing and in the management of the patients affected by alcohol related problems, such-as acute alcohol intoxication (AAI), alcohol withdrawal (AW), traumatic or spontaneous illness due to acute or chronic effects of alcohol abuse. Here are described both the clinical problems and the diagnostic instruments that nurse can use in ED to increase diagnostic sensibility. Furthermore both clinical features of AAI in relation with blood alcohol concentration (BAC), diagnostic criteria and test to evaluate AW severity are reported in this review. Even diagnostic instruments to identify alcohol abuse also in trauma are reported: the alcohol breath test, BAC and laboratory test. Even the Alcohol Use Disorders Identification Test (A.U.D.I.T.) questionnaire is reported to recognize the alcohol abuser. In conclusion acquiring a specific training, nurse can play a major role in the complex intervention net of alcoholics rehabilitation. PMID- 12645397 TI - [Depression in epilepsy: symptom or syndrome?]. AB - An occurrence of depression or depressive symptomatology has been reported in 30% of patients with epilepsy. Depression has been reported peri- and interictally. To make a differentiation may be difficult in patients with frequent seizures. However, complex partial seizures, particularly if are located on temporal lobe, appear to be etiologic factors, especially in men with left-sided epileptic foci. Depression is also more frequent in patients treated with polytherapy, particularly with phenobarbital and vigabatrin. The depression appears to be endogenous and has also been described in patients with temporal lobectomy. Underlying risk factors (genetic, metabolic, etc) and some psychosocial condition also play a part and may explain the increased rates of depression in patients with epilepsy. Treatment approaches include psychotherapy, rationalization of antiepileptic drug medication and antidepressant treatment. The use of antidepressant treatment, in these patients, still raises uncertainties because of the widespread persuasion that this drugs exacerbated seizures. This adverse event is relatively uncommon at therapeutic dosages, and its incidence with some of most frequently used antidepressant drugs is close to that of spontaneous seizures calculated for the general population, but the incidence may rise up to 30-40% after overdosage. On the basis of the data reported in literature, it appears fair to say that maprotiline and amoxapine show the greatest seizure risk, whereas trazodone, fluoxetine and fluvoxamine show the least. The data also showed that antidepressant drugs may display both convulsant and anticonvulsant effect and it is likely that the most important factor to assess the effect of a given antidepressant drug in terms of inhibition-excitation is drug dosage. Nevertheless, further studies are needed in this field, both to clarify the complex modulating effects of antidepressants on seizure threshold and to identify clearer and safer guidelines to manage the treatment of patients with epilepsy and concomitant depression. PMID- 12645398 TI - [Neurogenic dysphagia: physiology, physiopathology and rehabilitative treatment]. AB - Swallowing is both a voluntary than a reflex function. It consist in transporting feeding from mouth to the stomach. Swallowing function occurs with very frequency during the day and needs complex neuromuscular coordination. Several neurologic diseases determine swallowing disorders. Dysphagia, is the difficulty in swallowing. In slight disorders, swallowing function is sufficiently compensated, symptoms are few or absent. Sometimes the patient is able to compensate and obtains a safe deglutition. Rehabilitation of swallowing disorders is based on the assessment of all symptoms and troubles causing dysphagia and on the improvement of the specific disabilities. Rehabilitation is aimed to make patient able for a safe oral feeding. We can use classic specific physiotherapy, compensatory movements of head and neck, electrostimulation, and the chemical myotomia by botulinum toxin injection. PMID- 12645399 TI - Clones, parents, persons and the law in Australia. AB - Current understanding of cloning and artificial reproductive technology was reviewed. The results show that they are in contradiction with fundamental concepts upon which our society is built, such as person and parenthood. Analyses of some recent British and Australian laws show that attempts to protect the individual by regulating some biological technologies through carefully drafted legislation are bound to fail. PMID- 12645400 TI - [Retinal complications of posterior vitreous detachment: retrospective analysis of 3 clinical cases]. AB - Posterior vitreous detachment is an irreversible consequence of vitreal aging and will be encountered even more frequently as the population grows older. Because vitreous detachments are not always complete and clean, associated retinal complications may develop. Patients should be carefully examined at initial symptoms in order to diagnose and treat possible retinal injuries as soon as possible. Here we report a retrospective analysis of three different retinal complications of posterior vitreous detachment. PMID- 12645401 TI - [Non-obese obese-like subjects: a new patient to be treated with diet]. AB - Decreased insulin sensitivity patients, although normal-weight, are treated with a "prudent" hypocaloric diet and exercise, in order to reduce insulin resistance. Metabolic (glycolipidic) alterations can be easily diagnosed by combination of clinic and laboratory tests. A diet model aimed at controlling insulin levels and inducing a light weight loss is proposed. PMID- 12645402 TI - [Sepsis]. PMID- 12645403 TI - [AED (automatic external defibrillators) are coming!]. PMID- 12645404 TI - [Out-of-hospital cardiac arrest. Mechanisms and treatment with automated external defibrillator]. AB - In Denmark, approximately 4500 persons suffer yearly from an out-of-hospital cardiac arrest with mortality close to 100%. The principal arrhythmia is ventricular fibrillation, which can only be treated effectively with prompt external defibrillation. Automatic external defibrillators (AED) are small, portable, easily operated devices. They have documented high specificity and sensitivity. Moreover, biphasic automatic external defibrillators are at least as effective as traditional monophasic defibrillators. Survival rates with good neurological status as high as 60% have been reported. Better survival of out-of hospital cardiac arrest victims requires, however, improvements throughout the chain of survival, not only more automatic external defibrillators. Therefore, the health care system has to discuss thoroughly and solve important questions regarding organisation, logistics, education, and legal aspects in order to improve survival for out-of-hospital cardiac arrest victims. PMID- 12645405 TI - [Gated myocardial scintigraphy: simultaneous determination of heart perfusion and pump function]. PMID- 12645406 TI - [Defects in the surgical glove barrier. Single or double gloves]. AB - The purpose of wearing surgical gloves is to provide an antimicrobial barrier between the hands of the surgical staff and the tissues of the patient. Glove perforations are common, depending on the function of the staff member and the duration and nature of the surgery. In addition, gloves can hydrate, which is also a potential route of infection. The use of two pairs of gloves (double gloves) efficiently reduces both the perforation rate and the hydration of the glove barrier. Decreased dexterity and tactile sensitivity, however, are common objections, which keep many surgeons from wearing double gloves. Moreover, they seem to have little practical importance. Neither single nor double gloves have proved their effectiveness in terms of actual infections, but it seems rational to maintain a surgical glove barrier. Double gloves can be recommended as a simple means of improving the integrity of this barrier. PMID- 12645407 TI - [Risk factors of rheumatoid arthritis]. AB - Rheumatoid arthritis is a chronic inflammatory joint disease with an annual incidence rate between 25 and 50 per 100,000 individuals. Most likely, a combination of inherited and environmental factors, such as infections, reproductive or hormonal factors, and factors related to life style, is responsible for the disease. Future etiological studies should aim at providing a better insight into the interaction between different risk factors. PMID- 12645408 TI - [A retrospective analysis of the results of obstetric acupuncture at Frederiksberg Hospital]. AB - INTRODUCTION: The use of obstetric acupuncture in Denmark is increasing but its use in routine clinical practice has not been evaluated. MATERIAL AND METHODS: This retrospective uncontrolled study comprises 691 women who attended Frederiksberg Hospital during pregnancy and delivery. Their mean age was 30.5 +/- 4.3 year, mean parity 1.3 +/- 0.6 (71.8% were primiparous), and the mean gestational age was 39 +/- 3.7 weeks. A total of 563 of the women (81.5%) were in labour. The effect of acupuncture was evaluated by the pregnant women alone for the pregnancy-related indications and by both women and midwifes for labour related indications. RESULTS: The women had acupuncture based on 997 individual indications (mean 1.4 +/- 0.5). Twenty-two indications were used. 78.8% of all treatments were for pain relief or anxiety. The scores were highly correlated between women and midwives as 42.2 and 40.6%, respectively, indicated "full effect" and 33.3 and 33.4% indicated "some effect". The midwives' scores were independent of the women's age, parity, gestational age and indication(s). One possible side effect was observed (temporary dropfoot). DISCUSSION: In this study, routine obstetric acupuncture was perceived as equally effective by the women and midwifes. The method is simple, inexpensive and without significant adverse effects. It deserves a place in the midwives' armamentarium. PMID- 12645409 TI - [Obese pregnant women have complicated deliveries]. AB - INTRODUCTION: On the background of the increasing prevalence of overweight and obesity, we wanted to describe the deliveries among this group of women. MATERIAL AND METHODS: A prospective study of 798 pregnant women during 1 1/2 year. According to Body-Mass-Index (BMI) all referred women were divided into three groups considering their age and parity. A) 270 pregnant women with BMI 26-30. B) 132 pregnant women with BMI > 30. C) 320 pregnant women with BMI 20-25, the control group. RESULTS: The deliveries were induced at 9% and 11% in group A and group B, making a significant difference to the induction of 5% among the controls (p < 0.05). The most frequent reason for induction was graviditas prolongata. This was the case for 5% in group A and 6% in group B compared to 3% in group C. This was not significant (p = 0.24). The rates of cesarean section were 19% and 24% in group A and B, making a significant difference to the rate of 9% among the controls (p < 0.01). Disproportio foetopelvina and late decelerations were the most frequent reason for cesarean section during delivery. Disproportio foetopelvina discovered during delivery was the case for 5% in group A and 8% in group B compared to 1% in group C. This difference is significant with p < 0.05. DISCUSSION: Our study shows that obese and pregnant women have a high rate of induction and postdatism. They have a higher rate of cesarean section compared to normal weight women and get more frequently cesarean section during delivery because of disproportio foetopelvina. Health service professionals working with pregnant women should be aware of the overweight problems. PMID- 12645410 TI - [A nodule in the thyroid gland. Preoperative examinations and treatment--an analysis of 990 cases]. AB - INTRODUCTION: It is very difficult to know whether a nodule in the thyroid gland is benign or malignant as judged by a clinical examination, scintigraphy, US, or FNA. MATERIAL AND METHODS: Over a 10-year period a series of 1509 patients were operated on for diseases in the thyroid gland. The indication for surgical intervention in 990 patients was a nodule in the thyroid gland, defined as a solitary nodule or a suspicious nodule in a goitre. RESULTS: The incidence of malignancy in these 990 patients with a nodule was 6.3% with no difference between men and women. Forty-four per cent were follicular adenomas and the rest were colloid nodules. No increased incidence of malignancy in nodules was found with increasing size, whereas the incidence significantly increased with advancing age. Scintigraphy was carried out in almost all the patients (98.9%). Seventy-five per cent had a cold nodule, 9% irregular uptake, and 17% a warm nodule. Only one malignancy, a follicular carcinoma, was a warm nodule. FNA was performed in 955 of the 990 patients (97%). The incidence of follicular neoplasia was 33% in the benign nodule and 70% in the follicular adenoma. DISCUSSION: When malignant cells were found by FNA, it was an almost certain sign of malignancy with very few false positive results, whereas a follicular adenoma or a malignant tumour cannot be excluded by a negative answer. The risk of complications was very low, with an 0.3% incidence of recurrent nerve palsy. If and when the risk can be kept at a reasonably low level, the advantages of thyroid surgery will out number the disadvantages and early surgery is therefore preferable. PMID- 12645411 TI - [Vigabatrin and visual field defects. A Danish material with evaluation of different screening methods]. AB - INTRODUCTION: The aim of the study was to assess the prevalence of visual field constriction in a consecutive, unselected series of patients in the Copenhagen area treated with vigabatrin for severe epilepsy, and further to assess appropriate screening techniques for detecting such visual field defects. MATERIAL AND METHODS: During 1999, 36 vigabatrin-treated patients (aged 13-67) referred to the eye clinic, had a full ophthalmic examination. Routine visual field testing by means of a) finger movements and b) a tangent screen was supplemented by c) manual kinetic Goldmann perimetry and d) automated static threshold profile perimetry (Octopus). RESULTS: Of the 31 subjects accepted for entry, 80% showed some degree of visual field constriction. The affection was regarded as slight in 15 cases, moderate in seven, and severe in four. Both kinetic Goldmann and static computerised perimetry disclosed the peripheral defects in those affected. By Goldmann perimetry, it appeared as a smooth narrowing of outer visual field borders, whereas Octopus demonstrated a less regular, more scattered loss of sensitivity. DISCUSSION: Eighty per cent with peripheral field affection is among the highest reported frequencies in a vigabatrin-treated series of patients so far. An association with a cumulated dose of the drug was demonstrated. Visual field testing by finger movements and tangent screen evaluation will not reveal the initial peripheral losses of the visual field. For this purpose, we found manual kinetic Goldmann perimetry a valid alternative to modern computerised perimetric techniques. Goldmann takes less time, makes the co-operation of the patient easier, and the interpretation is simple. PMID- 12645412 TI - [Laparoscopic wedge resection of stromal gastric tumor guided by laparoscopic ultrasonographic scanning]. AB - A case of stromal gastric tumor in a 64-year-old man treated by laparoscopic wedge resection guided by laparoscopic ultrasonography is described. The procedure seems very suitable because it combines the advantages of minimal surgery with the application of ultrasonography, which compensates for the lack of tactile sensitivity in laparoscopic surgery. PMID- 12645413 TI - [Splenic cysts--pathogenesis, diagnostics and treatment]. AB - A case of a benign noninfectious cyst is reported. Splenic cysts are uncommon. They are diagnosed by ultrasonography, computer tomography, and nuclear magnetic resonance. The cysts are either infectious (e.g. parasitic) or noninfectious (e.g. posttraumatic). The cystic content should be analysed for bacteria, parasites, and amylase. Noninfectious cysts larger than four cm and/or symptomatic cysts should be treated surgically with laparoscopic decapsulation. PMID- 12645414 TI - [Life-threatening angioedema associated with ACE inhibitor treatment]. AB - A case of severe late-onset angioedema with life-threatening upper airway obstruction, related to ACE inhibitor treatment is presented. The patient was a middle-aged man treated for hypertension, presenting with stridor requiring intubation, which was difficult. The patient developed cardiac arrest but was successfully resuscitated, after intubation. Etiology, risk factors, and treatment, with special attention to air way management, are discussed. PMID- 12645415 TI - [Acute renal failure in intensive care units--evidence and cost-benefits when choosing the method of dialysis]. PMID- 12645416 TI - [Calculated consequences of the Danish treatment of Staphylococcus aureus bacteremia and meningitis]. PMID- 12645417 TI - [Why meropenem and not imipenem?]. PMID- 12645418 TI - [Disinfection]. PMID- 12645419 TI - Health education and food safety behavior in the university setting. AB - Health education in colleges and universities offers an opportunity to reach a captive audience of young people in order to promote a lifetime of healthy behavior. University health education typically focuses on topics such as alcohol and substance abuse, sexual behavior, and nutrition. Environmental health issues are not a prominent component of university health education, even though an understanding of a healthy environment is critical to promoting overall health. Food safety is an environmental health issue of particular concern in universities since many young adults prepare their own meals for the first time there. Food safety questions from the Centers for Disease Control and Prevention's Behavioral Risk Factor Surveillance Survey were used to assess the behaviors of 354 upper-level undergraduate students. Findings from the survey indicate that students are engaging in risky food consumption and handling behaviors and that the educational techniques mandated by the federal government are ineffective in reaching college students. PMID- 12645420 TI - Skills, knowledge, and abilities of graduates from accredited environmental health science and protection undergraduate programs. AB - The National Environmental Health Science and Protection Accreditation Council (EHAC) has the mission of enhancing the education and training of students who intend to become environmental health science and protection practitioners/professionals. Academic programs that demonstrate compliance with EHAC guidelines can become accredited with the expectation that graduates of accredited programs will have the knowledge, skills, and abilities needed for professional success. The study reported on here reviewed these guidelines in light of results from a survey of program graduates and their supervisors. The survey results were found to establish that the current approach is quite successful, and that graduates of the programs have the potential to make substantial professional contributions in protecting environmental health. PMID- 12645421 TI - Every kitchen's dirty little secret: cardboard--bacteria superstore and roach motel. PMID- 12645422 TI - Fecal sterols: the next-generation sewage indicator. PMID- 12645423 TI - Employees and neighbors. PMID- 12645424 TI - Better plan needed to protect U.S. agriculture from bioterrorism. PMID- 12645426 TI - Additives linked to behavior. PMID- 12645425 TI - Checking on changes: consumer research. PMID- 12645427 TI - How NEHA can help you in a down economy--a specific example. PMID- 12645428 TI - Charity stops at the doctor's door. PMID- 12645429 TI - Stopped at the border. PMID- 12645430 TI - Local employers: they're worth getting to know. PMID- 12645431 TI - Cancer programs get a boost. PMID- 12645432 TI - Choose your words wisely. PMID- 12645433 TI - Generic drugs get a life. PMID- 12645434 TI - Cultivating patient relationships. PMID- 12645435 TI - What's missing from patient-centered care? PMID- 12645436 TI - Healthcare warms up to the Web. Interview by Richard K. Thomas. PMID- 12645437 TI - Guerrilla e-health. It takes aggressive management to make a Web site work. PMID- 12645438 TI - When bad things happen to good hospitals. How one facility rose above a publicity crisis. PMID- 12645439 TI - Living with abundance. Disciplined marketing will avoid waste and bring success. PMID- 12645440 TI - Locomotor training in neurorehabilitation: emerging rehabilitation concepts. PMID- 12645441 TI - Effectiveness of physiotherapy, occupational therapy, and speech pathology for people with Huntington's disease: a systematic review. AB - This review provides a summary of the current literature examining the outcomes of physiotherapy, occupational therapy, and speech pathology interventions for people with Huntington's disease. The literature was retrieved via a systematic search using a combination of key words that included Huntington's disease, physiotherapy, occupational therapy, and speech pathology. The electronic databases for Medline, Embase, CINAHL, Cochrane Controlled Trials Register, and PEDro were searched up to May 2002. Articles meeting the review criteria were graded for study type and rated for quality using checklists to assess study validity and methodology. The majority of articles that examined therapy outcomes for people with Huntington's disease were derived from observational studies of low methodological quality. A low level of evidence exists to support the use of physiotherapy for addressing impairments of balance, muscle strength, and flexibility. There was a small amount of evidence to support the use of speech pathology for the management of eating and swallowing disorders. The current evidence is insufficient to make strong recommendations regarding the usefulness of physiotherapy, occupational therapy, or speech pathology for people with Huntington's disease. There is further need for therapy outcomes research in Huntington's disease so that clinicians may use evidence-based practice to assist clinical decision making. PMID- 12645442 TI - Kinematic adaptations of spinal cord-injured subjects during obstructed walking. AB - STUDY DESIGN: A case-control study of walking over obstacles. OBJECTIVE: To characterize and compare the kinematic, anticipatory locomotor adjustments used by people with incomplete spinal cord injuries (SCIs). METHODS: The angular and linear kinematics of the lower limb when going over obstacles of low height (0.005 and 0.030 m) were compared between SCI subjects (n = 6) and able-bodied participants (n = 5). RESULTS: The results of this study show that even though SCI participants could adapt their kinematic patterns to go over obstacles, none of the participants used kinematic strategies similar to those of able-bodied participants. CONCLUSIONS: This difference could be explained in part by the absence in the SCI participants of increased hip flexion when going over the obstacles. Other confounding factors are discussed. PMID- 12645443 TI - Respiratory management of pediatric patients with spinal cord injuries: retrospective review of the duPont experience. AB - Pulmonary complications contribute to morbidity and mortality in spinal cord injuries (SCIs). A retrospective review of 20 years of experience with tracheostomy- and ventilator-dependent SCI children is presented. The authors developed and analyzed a database of 47 children (average age = 11.4 years). Of the patients, 27% had concomitant brain injuries, 6% had prior histories of reactive airway disease, and 2% had thoracic fractures. Injuries were caused by motor vehicle accidents (53%); gunshot wounds (19%); sports-related accidents (19%); and vascular injuries, transverse myelitis, or spinal tumors (8%). Of the injuries, 52% were high level (C1 to C2) and 48% were mid- or low level (C3 to C5). Two groups were analyzed for demographic information. Complications included tracheitis, atelectasis, and pneumonia. Mean tidal volume was 14 cm2/kg (maximum = 22 cm2/kg). Bedside lung function parameters were attempted to assess readiness and the rapidity of weans. T-piece sprints were used to successfully wean 63% of patients. Successfully weaned patients were compared with those not weaned. No deaths or readmissions for late-onset respiratory failure postwean occurred. The authors' clinical impression favors higher tidal volumes and aggressive bronchial hygiene to minimize pulmonary complications and enhance weaning. Successfully weaned patients had fewer complications. A critical pathway for respiratory management of SCI children is presented. PMID- 12645444 TI - An fMRI investigation of hand representation in paraplegic humans. AB - OBJECTIVE: Cortical reorganization can occur after deaf-ferentation due to loss of a limb, but the nature of the cortical reorganization after spinal cord injury (SCI) is still in debate. METHODS: Using a 1.5T MRI, we scanned paraplegic and noninjured participants during hand movement and palm stimulation, to determine whether longterm paraplegics would show different patterns of cortical activity from the noninjured participants. RESULTS: The SCI group showed stronger activation in areas posterior, rather than superior, to the areas activated by non-SCIs. Conversely, the non-SCIs showed stronger activation in more anterior areas. The signal at each individual's maximally significant voxel had a greater modulation for the SCI group than for the non-SCIs, in response to movement. CONCLUSIONS: In this study of sensory and motor representations within the same subjects, the authors show for the first time the increase in the BOLD fMRI signal modulation in SCI. The authors do not find evidence of expansion of the hand representation into nearby cortical areas, and they corroborate previous EEG studies indicating a posterior shift for hand motor representation after SCI, while showing that the sensory representation does not undergo a posterior shift of similar magnitude. The difference between the reorganization found here and the reorganization typically found following amputations suggests a rationale for the differences in neuropathic pain symptoms following a spinal cord injury or amputation. PMID- 12645446 TI - Sensory reinnervation of myocutaneous flaps revealed by infrared laser evoked sensations and brain potentials. AB - The aim of this study was to examine the recovery of sensory function in myocutaneous flaps comparing 2 test methods. Eight flaps in 7 patients were examined by using clinical neurological test procedures (CNT) in comparison with psychophysics and evoked brain potentials (LEP) following infrared laser stimuli. The authors found that only 3 out of 8 flaps in 7 patients exhibited signs of reinnervation when tested with CNT. Three grades of reinnervation appeared in 7 flaps when tested with the laser. Grade 1 indicated the recovery of unmyelinated C-fiber function in 7 flaps accounting for the ability to discriminate laser intensities by different degrees of warmth. Grade 2 appeared in 3 of these flaps and was characterized by the additional ability to sense pinprick pain and the elicitation of late components of LEP mediated by thinly myelinated A delta nociceptors. Grade 3 involved the additional sensibility for superficial touch indicating the recovery of thickly myelinated A beta-fibers noted in 2 of these flaps. The authors conclude that the LEP method is more sensitive than standard neurological test procedures to objectively document early signs of reinnervation after reconstructive flap surgery. This result is promising to investigate greater patient populations comparing different surgical techniques in future studies. PMID- 12645445 TI - Constraint-induced therapy in stroke: magnetic-stimulation motor maps and cerebral activation. AB - Constraint-induced movement therapy (CI), a standardized intensive rehabilitation intervention, was given to patients a year or more following stroke. The goal was to determine if CI was more effective than a less-intensive control intervention in changing motor function and/or brain physiology and to gain insight into the mechanisms underlying this recovery process. Subjects were recruited and randomized more than 1 year after a single subcortical infarction. Clinical assessments performed before and after the intervention and at 6 months postintervention included the Wolf Motor Function Test (WMFT), the Motor Activity Log (MAL), and the Assessment of Motor and Process Skills (AMPS). Transcranial magnetic stimulation was used to map the motor cortex. Positron emission tomography was used to measure changes in motor task-related activation due to the intervention. MAL increased by 1.08 after CI therapy and decreased by 0.01 after control therapy. The difference between groups was significant (P < 0.001). Changes in WMFT and AMPS were not significantly different between groups. Cerebral activation during a motor task decreased significantly, and motor map size increased in the affected hemisphere motor cortex in CI patients but not in control patients. Both changes may reflect improved ability of upper motor neurons to produce movement. PMID- 12645447 TI - Spasticity in multiple sclerosis. AB - The objective of this article is to establish the prevalence of spasticity in a random selection of people with multiple sclerosis (MS) in the city of Newcastle upon Tyne in the Northeast of England. A secondary aim was to assess the adequacy of current pharmacological intervention for spasticity and assess the relationship between spasticity and overall disability. The study design was a simple comparison that examined differences in functional independence in 2 random groups of people with MS subdivided by the presence of clinically significant spasticity. A total of 68 adults with a diagnosis of clinically definite MS were included in the study. Their level of functional independence was assessed using the Newcastle Independence Assessment Form (NIAF), the Functional Independence Measure (FIM), and the Kurtzke Extended Disability Status Scale (EDSS). Spasticity was assessed using the Modified Ashworth Scale. A subjective analysis was made of the appropriateness of oral antispastic medication by a rehabilitation physician. Thirty-two people (47%) had clinically significant spasticity (Modified Ashworth Score of 2, 3, or 4). Seventy-eight percent of the population were receiving some oral antispastic medication, but 50% were deemed to require some drug adjustment or additional treatment. Individuals with spasticity were found to have significantly higher levels of disability than those who had no spasticity or clinically insignificant spasticity. This study has confirmed that spasticity is highly prevalent in the MS population and is significantly associated with a reduced level of functional independence. Treatment of spasticity is suboptimal in a large proportion of the population, and the need for further information and education to health professionals and to people with MS is highlighted. PMID- 12645448 TI - Highlighting 2002 award-winning initiatives. AB - This issue takes an instructive look at four organization--SSM Health Care (based in St. Louis), Children's Hospital and Health Center of San Diego, Concord (NH) Hospital, and McLeod (SC) Regional Medical Center--that received top honors this past year for their ongoing efforts to provide quality healthcare. PMID- 12645449 TI - New IOM report targets 20 priority areas to improve healthcare quality. AB - A new Institute of Medicine report lists 20 priority areas with "the greatest opportunities for closing the gap between known best practice and usual practice" to reduce unwanted variation and promote quality healthcare. PMID- 12645450 TI - A precarious position in turbulent times. PMID- 12645451 TI - [Margot Phaneuf, from vocation to profession. Interview by Karine Fortin]. PMID- 12645452 TI - [Living with asthma (part 2). Essential information for asthma control]. PMID- 12645453 TI - [Surpassing oneself in order to cope... The therapeutic adventure]. PMID- 12645454 TI - [Why not? Similarities and differences in social leadership in the heart of the nursing profession in Montreal and Rio]. PMID- 12645455 TI - [Hold on tight, I will help you!]. PMID- 12645456 TI - [Health without smoke]. PMID- 12645457 TI - [Systematic reflective approach for whom? Why?]. PMID- 12645458 TI - Consumer health library Websites--great marketing tools. PMID- 12645459 TI - From a past chair--how do hospital librarians in HLS and MLA get into offices and on committees? PMID- 12645461 TI - Things aren't getting better. Experts at federal hearing say healthcare costs, quality and access haven't improved much, if at all, since managed care. AB - Nearly a decade after managed care was supposed to revolutionize the healthcare industry, Americans have enjoyed little to no improvement in healthcare costs, quality or access, experts testified at a federal hearing last week. The hearing, arranged by the U.S. Justice Department and the Federal Trade Commission, was the first in a series of meetings. PMID- 12645460 TI - Spoonfeeding the masses. PMID- 12645463 TI - Radical departure. Reform group common good wants medical experts to replace juries in malpractice cases. PMID- 12645462 TI - Consolidation squabble. AHA study refutes Blues on cause for rising costs. PMID- 12645464 TI - Medicaid melee. Governors snub Bush's reform proposals. PMID- 12645465 TI - Adventures in joint ventures. Texas hospital will be Triad's first with a not-for profit. PMID- 12645466 TI - Dial-a-doc gets boost. JCAHO issues first official telemedicine endorsement. PMID- 12645468 TI - Keeping up to date. PMID- 12645467 TI - Banner's end run must end. Company plays chess with assets it inherited while communities pay the price. PMID- 12645469 TI - Still leader of the pack. Intermountain tops ranking of integrated networks. PMID- 12645470 TI - Intensive caring. He's a physician-lawyer-consultant, and his patients are typically small hospitals on life support. PMID- 12645471 TI - Big, bad brutes? PMID- 12645472 TI - A better start? PMID- 12645473 TI - Protection for protesters. PMID- 12645474 TI - Lessons for battle. A failed AIDS vaccine trial sheds new light on the disease. PMID- 12645475 TI - Pinpoint fat relief? PMID- 12645476 TI - Taming killer clots. PMID- 12645477 TI - The doctor wore a dive mask. PMID- 12645478 TI - Keeping details from the devil. PMID- 12645479 TI - Casting and strapping rules. PMID- 12645480 TI - Practice management systems. Here's what to look for. PMID- 12645481 TI - Don't play solitaire, and other office computer rules. PMID- 12645482 TI - Let employees know where they stand. PMID- 12645483 TI - Who taught you what medicine is? PMID- 12645484 TI - You've been sued. There's help. PMID- 12645485 TI - Missed the HIPAA deadline? Now what. PMID- 12645486 TI - Moves you shouldn't make. PMID- 12645487 TI - Hire right, or pay the price. PMID- 12645488 TI - RNA silencing: a new therapeutic strategy against HIV. PMID- 12645489 TI - New year, new Congress, same old concerns. PMID- 12645490 TI - Musculoskeletal manifestations of HIV infection. PMID- 12645491 TI - Merging risk management and quality assurance in ambulatory HIV care: part 1, the process. PMID- 12645492 TI - Improving adherence to antiretroviral therapy. AB - Taking antiretroviral medications as directed is essential to reducing the plasma viral load. By controlling HIV replication, antiretroviral combination therapy enables the host's immune system to fortify itself and ward off opportunistic pathogens. However, adhering to antiretroviral regimens that include at least 3 different drugs taken once, twice, or 3 times a day is a constant challenge for patients with HIV disease. Care providers need to be aware of the challenges and obstacles faced by their patients on antiretroviral regimens, and efforts to maximize adherence must begin even before the patient has the first prescriptions filled. PMID- 12645493 TI - Secondary erythermalgia in an HIV-1-positive patient. AB - This report describes an HIV-1-positive patient who presented to the dermatology clinic with a 2-year history of secondary erythermalgia. A review of salient clinical features, disease classification, associated diseases, pathogenesis, and treatment of this disorder is presented. PMID- 12645494 TI - Education for continuous subcutaneous insulin infusion pump users. AB - Successful implementation of CSII requires a motivated patient with a range of technical skills and self-management capabilities. Patients develop this expertise through an ongoing program of education and the support that a healthcare team knowledgeable in insulin pump therapy can provide. PMID- 12645495 TI - How did the introduction of managed care for the uninsured in Iowa affect the use of substance abuse services? AB - Concerns about access under managed care have been raised for vulnerable populations such as publicly funded patients with substance abuse problems. To estimate the effects of the Iowa Managed Substance Abuse Care Plan (IMSACP) on substance abuse service use by publicly funded patients, service use before and after IMSACP was compared; adjustments were made for changes in population sociodemographic and clinical characteristics. Between fiscal years 1994 and 1997, patient case mix was marked by a higher burden of illness and the use of inpatient, residential nondetox, outpatient counseling, and assessment services declined, while use of intensive outpatient and residential detox services increased. Findings were similar among women, children, and homeless persons. Thus, care moved away from high-cost inpatient settings to less costly venues. Without knowing the impact on treatment outcomes, these changes cannot be interpreted as improved provider efficiency versus simply cost containment and profit maximization. PMID- 12645497 TI - Report of the Coding Committee of the Association for European Paediatric Cardiology. PMID- 12645496 TI - Does age at the time of elective cardiac surgery or catheter intervention in children influence the longitudinal development of psychological distress and styles of coping of parents? AB - AIMS: To assess the influence of age at a cardiac procedure of children, who underwent elective cardiac surgery or interventional cardiac catheterisation for treatment of congenital cardiac defects between 3 months and 7 years of age, on the longitudinal development of psychological distress and styles of coping of their parents. METHODS: We used the General Health Questionnaire to measure psychological distress, and the Utrecht Coping List to measure styles of coping. Parents completed questionnaires on average respectively 5 weeks prior to, and 18.7 months after, cardiac surgery or catheter intervention for their child. RESULTS: Apart from one exception, no significant influence was found of the age at which children underwent elective cardiac surgery or catheter intervention on the pre- to postprocedural course of psychological distress and the styles of coping of their parents. Across time, parents of children undergoing surgery reported, on average, significantly higher levels of psychological distress than parents of children who underwent catheter intervention. After the procedure, parents of children who underwent either procedure reported significantly lower levels of psychological distress, and showed a weaker tendency to use several styles of coping, than did their reference groups. CONCLUSION: Age of the children at the time of elective cardiac surgery or catheter intervention did not influence the course of psychological distress of their parents, nor the styles of coping used by the parents. Future research should investigate in what way the age at which these cardiac procedures are performed influences the emotional and cognitive development of the children. PMID- 12645498 TI - 3D bulk measurements of the force distribution in a compressed emulsion system. AB - In particulate materials, such as emulsions and granular media, a "jammed" system results if particles are packed together so that all particles are touching their neighbours, provided the density is sufficiently high. This paper studies through experiment, theory and simulation, the forces that particles exert upon one another in such a jammed state. Confocal microscopy of a compressed polydisperse emulsion provides a direct 3D measurement of the dispersed phase morphology within the bulk of the sample. This allows the determination of the probability distribution of interdroplet forces, P(f) where f is the magnitude of the force, from local droplet deformations. In parallel, the simplest form of the Boltzmann equation for the probability of force distributions predicts P(f) to be of the form e(-f/p), where p is proportional to the mean force f for large forces. This result is in good agreement with experimental and simulated data. PMID- 12645500 TI - Thermometer exchange tips. PMID- 12645499 TI - Smallpox Facts. PMID- 12645502 TI - [Use, clinical testing and safety of medical products and the reporting of incidents from the physicians's viewpoint]. PMID- 12645503 TI - [Surgical treatment of dissecting aneurysm of the descending aorta: report of one case]. PMID- 12645504 TI - [Sudden onset of unilateral right-sided symptoms. Unilateral venous infarct of the left thalamus in thrombosis of the internal cerebral veins]. PMID- 12645505 TI - [Improving the control of systolic hypertension and pulse pressure in elderly population]. PMID- 12645510 TI - [Case no 1: pulmonary pathology. Congenital atresia of the apical posterior segment of the left pulmonary lobe]. PMID- 12645511 TI - [Case no 2. Abscess due to sigmoid diverticulum perforation]. PMID- 12645509 TI - [Quid? Jejunal diverticulitis with intra-abdominal abscess and reactional jejunitis]. PMID- 12645512 TI - [Case no 3: pulmonary and hepatic pathology]. PMID- 12645513 TI - [Case no 4. Aggressive fibromatosis (desmoid tumor) associated with surgically treated familial colonic and duodenal polyposis]. PMID- 12645514 TI - [Case no 5. Fasciola hepatica distomatosis]. PMID- 12645515 TI - [Case no 6. Benin lipoma]. PMID- 12645516 TI - [Case no 1. Neuroepithelial dysembryoplastic tumor]. PMID- 12645518 TI - [Case no 2. Balo concentric sclerosis]. PMID- 12645517 TI - Acute phase mediators modulate thrombin-activable fibrinolysis inhibitor (TAFI) gene expression in HepG2 cells. AB - Thrombin-activable fibrinolysis inhibitor (TAFI) has recently been identified as a positive acute phase protein in mice, an observation that may have important implications for the interaction of the coagulation, fibrinolytic, and inflammatory systems. Activated TAFI (TAFIa) inhibits fibrinolysis by removing the carboxyl-terminal lysines from partially degraded fibrin that are important for maximally efficient plasminogen activation. In addition, TAFIa has been shown to be capable of removing the carboxyl-terminal arginine residues from the anaphylatoxins and bradykinin, thus implying a role for the TAFI pathway in the vascular responses to inflammation. In the current study, we investigated the ability of acute phase mediators to modulate human TAFI gene expression in cultured human hepatoma (HepG2) cells. Surprisingly, we found that treatment of HepG2 cells with a combination of interleukin (IL)-1 and IL-6 suppressed endogenous TAFI mRNA abundance in HepG2 cells (~60% decrease), while treatment with IL-1 or IL-6 alone had no effect. Treatment with IL-1 and/or IL-6 had no effect on TAFI promoter activity as measured using a luciferase reporter plasmid containing the human TAFI 5'-flanking region, whereas treatment with IL-1 and IL 6 in combination, but not alone, decreased the stability of the endogenous TAFI mRNA. Treatment with the synthetic glucocorticoid dexamethasone resulted in a 2 fold increase of both TAFI mRNA levels and promoter activity. We identified a functional glucocorticoid response element (GRE) in the human TAFI promoter between nucleotides 92 and 78. The GRE was capable of binding the glucocorticoid receptor, as assessed by gel mobility shift assays, and mutation of this element markedly decreased the ability of the TAFI promoter to be activated by dexamethasone. PMID- 12645519 TI - [Case no 3. MELAS syndrome]. PMID- 12645520 TI - [Case no 4. Intracranial hypotension syndrome]. PMID- 12645521 TI - [Case no 5. Wilson's disease]. PMID- 12645523 TI - [Case no 6. Cerebral vasculitis due to Toxocara canis (or catis) origin]. PMID- 12645522 TI - The C2A domain of synaptotagmin alters the kinetics of voltage-gated Ca2+ channels Ca(v)1.2 (Lc-type) and Ca(v)2.3 (R-type). AB - Biochemical and genetic studies implicate synaptotagmin (Syt 1) as a Ca2+ sensor for neuronal and neuroendocrine neurosecretion. Calcium binding to Syt 1 occurs through two cytoplasmic repeats termed the C2A and C2B domains. In addition, the C2A domain of Syt 1 has calcium-independent properties required for neurotransmitter release. For example, mutation of a polylysine motif (residues 189-192) reverses the inhibitory effect of injected recombinant Syt 1 C2A fragment on neurotransmitter release from PC12 cells. Here we examined the requirement of the C2A polylysine motif for Syt 1 interaction with the cardiac Cav1.2 (L-type) and the neuronal Cav2.3 (R-type) voltage-gated Ca2+ channels, two channels required for neurotransmission. We find that the C2A polylysine motif presents a critical interaction surface with Cav1.2 and Cav2.3 since truncated Syt 1 containing a mutated motif (Syt 1*1-264) was ineffective at modifying the channel kinetics. Mutating the polylysine motif also abolished C2A binding to Lc753-893, the cytosolic interacting domain of Syt 1 at Cav1.2 1 subunit. Syt 1 and Syt 1* harboring the mutation at the KKKK motif modified channel activation, while Syt 1* only partially reversed the syntaxin 1A effects on channel activity. This mutation would interfere with the assembly of Syt 1/channel/syntaxin into an exocytotic unit. The functional interaction of the C2A polylysine domain with Cav1.2 and Cav2.3 is consistent with tethering of the secretory vesicle to the Ca2+ channel. It indicates that calcium-independent properties of Syt 1 regulate voltage-gated Ca2+ channels and contribute to the molecular events underlying transmitter release. PMID- 12645524 TI - [To make greater brilliance based on the one-century glory--pleasant retrospect and excellent prospect of electrocardiography. ]. PMID- 12645525 TI - [Pay great attention to early diagnosis and treatment of breast cancer]. PMID- 12645526 TI - Functional activity of the complement regulator encoded by Kaposi's sarcoma associated herpesvirus. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is closely associated with Kaposi's sarcoma and certain B-cell lymphomas. The fourth open reading frame of the KSHV genome encodes a protein (KSHV complement control protein (KCP, previously termed ORF4)) predicted to have complement-regulating activity. Here, we show that soluble KCP strongly enhanced the decay of classical C3-convertase but not the alternative pathway C3-convertase, when compared with the host complement regulators: factor H, C4b-binding protein, and decay-accelerating factor. The equilibrium affinity constant (KD) of KCP for C3b and C4b was determined by surface plasmon resonance analysis to range between 0.47-10 microM and 0.025-6.1 microM, respectively, depending on NaCl concentration and cation presence. Soluble and cell-associated KCP acted as a cofactor for factor I (FI) mediated cleavage of both C4b and C3b and induced the cleavage products C4d and iC3b, respectively. In the presence of KCP, FI further cleaved iC3b to C3d, which has never been described before as complement receptor 1 only mediates the production of C3dg by FI. KCP would enhance virus pathogenesis through evading complement attack, opsonization, and anaphylaxis but may also aid in targeting KSHV to one of its host reservoirs since C3d is a ligand for complement receptor 2 on B-cells. PMID- 12645528 TI - Quantitative assessment of fracture healing using positron emission tomography. PMID- 12645527 TI - Platelet-derived growth factor induces the beta-gamma-secretase-mediated cleavage of Alzheimer's amyloid precursor protein through a Src-Rac-dependent pathway. AB - The beta-amyloid peptide (Abeta) present in the senile plaques of Alzheimer's disease derives from the cleavage of a membrane protein, named APP, driven by two enzymes, known as beta- and gamma-secretases. The mechanisms regulating this cleavage are not understood. We have developed an experimental system to identify possible extracellular signals able to trigger the cleavage of an APP-Gal4 fusion protein, which is detected by measuring the expression of the CAT gene transcribed under the control of the Gal4 transcription factor, which is released from the membrane upon the cleavage of APP-Gal4. By using this assay, we purified a protein contained in the C6 cell-conditioned medium, which activates the cleavage of APP-Gal4 and which we demonstrated to be PDGF-BB. The APP-Gal4 processing induced by PDGF is dependent on the gamma-secretase activity, being abolished by an inhibitor of this enzyme, and is the consequence of the activation of a pathway downstream of the PDGF-receptor, which includes the non receptor tyrosine kinase Src and the small G-protein Rac1. These findings are confirmed by the observation that a constitutively active form of Src increases Abeta generation and that, in cells stably expressing APP, the generation of A is strongly decreased by the Src tyrosine kinase inhibitor PP2. PMID- 12645531 TI - Bibliography. Current world literature. Epidemiology and health-related services. PMID- 12645529 TI - Interferon-gamma induces p11 gene and protein expression in human epithelial cells through interferon-gamma-activated sequences in the p11 promoter. AB - The effect of interferon (IFN)-gamma on p11 expression was studied in two human epithelial cell lines (BEAS-2B and HeLa). Treatment with IFN-gamma resulted in increased steady-state levels of p11 mRNA and protein expression, with a time dependent and dose-dependent effect. Transient transfection experiments of a reporter gene construct containing 1498 bp of the 5'-flanking region of the p11 promoter demonstrated that IFN-gamma induced p11 gene expression at the transcriptional level. These effects were inhibited at the promoter and protein levels by a specific JAK-2 kinase inhibitor, AG-490. Functional analysis of the p11 promoter indicates that two gamma-activated sequence elements (GAS) located at positions 1219 and 1090 are important for the induction of the p11 promoter by IFN-gamma. Transfection of mutated reporter constructs demonstrated that the mutation at the GAS-2 site (1090) inhibited the p11 promoter activity, with a reduction of about approximately 73% and mutation at the GAS-3 site (1219) eliminated about 26% of the p11 promoter activity. A STAT1 dominant negative mutant vector at Tyr-701 (JAK kinase phosphorylation site) blocked the effect of IFN-gamma on the p11 promoter activity. IFN-gamma induced a rapid tyrosine phosphorylation and nuclear translocation of STAT1 protein, which is involved in the binding to the GAS-2 site in the p11 promoter by EMSA analysis. These data suggest that IFN-gamma-induced p11 expression is mediated through the binding of STAT1 to GAS sites in the p11 promoter. Inhibition of p11 expression by inhibitory antisense RNAs (iRNA) treatment resulted in enhanced IFN-gamma and calcium ionophore-stimulated arachidonic acid release suggesting that at least in part IFN-gamma-stimulated p11 expression may serve a counterregulatory role. PMID- 12645530 TI - ATP activates ataxia-telangiectasia mutated (ATM) in vitro. Importance of autophosphorylation. AB - Ataxia-telangiectasia Mutated (ATM), mutated in the human disorder ataxia telangiectasia, is rapidly activated by DNA double strand breaks. The mechanism of activation remains unresolved, and it is uncertain whether autophosphorylation contributes to activation. We describe an in vitro immunoprecipitation system demonstrating activation of ATM kinase from unirradiated extracts by preincubation with ATP. Activation is both time- and ATP concentration-dependent, other nucleotides fail to activate ATM, and DNA is not required. ATP activation is specific for ATM since it is not observed with kinase-dead ATM, it requires Mn2+, and it is inhibited by wortmannin. Exposure of activated ATM to phosphatase abrogates activity, and repeat cycles of ATP and phosphatase treatment reveal a requirement for autophosphorylation in the activation process. Phosphopeptide mapping revealed similarities between the patterns of autophosphorylation for irradiated and ATP-treated ATM. Caffeine inhibited ATM kinase activity for substrates but did not interfere with ATM autophosphorylation. ATP failed to activate either A-T and rad3-related protein (ATR) or DNA-dependent protein kinase under these conditions, supporting the specificity for ATM. These data demonstrate that ATP can specifically induce activation of ATM by a mechanism involving autophosphorylation. The relationship of this activation to DNA damage activation remains unclear but represents a useful model for understanding in vivo activation. PMID- 12645532 TI - Bibliography. Current world literature. Rehabilitation medicine in rheumatic diseases. PMID- 12645534 TI - Practical bioinformatics. PMID- 12645533 TI - Uses of error: Surgical mistakes. PMID- 12645535 TI - Venous thrombosis complicating inferior vena cava anomalies in a 7-year-old boy. PMID- 12645536 TI - BSE in human tissues fires debate on patient disclosure. PMID- 12645537 TI - Morphogenesis: clinical natural history and imaging information on patients included in reports. PMID- 12645538 TI - [Collagenous colitis: report of one case]. PMID- 12645539 TI - Acute appendicitis perforated into the bladder. PMID- 12645540 TI - Hospitalists becoming a popular practice. PMID- 12645541 TI - Loyalty to whom? PMID- 12645542 TI - Bibliography. Current world literature. Clinical nephrology. PMID- 12645543 TI - Bibliography. Current world literature. Pathophysiology of hypertension. PMID- 12645544 TI - [Ectopic thyroid: report of one case]. PMID- 12645545 TI - What do Ontario children eat for breakfast? Food group, Energy and macronutrient intake. AB - This study included 305 children living in Ontario in 1993. Our objective was to determine the proportion of daily energy and macronutrient intake consumed at breakfast, and the major food groups contributing to this meal. Demographic data were obtained in a parent interview that was part of the prevention project Better Beginnings, Better Futures. A single 24-hour recall among parents indicated that breakfast provided a mean of 1,230 (+/- 607) kJ. Although only 4.9% (n= 15) of children ate nothing at breakfast, 26.9% had <837 kJ. Many (59.7%) had amid-morning snack; however, children who consumed <837 kJ at breakfast were not more likely to have a snack than were those who had a greater energy intake. The major sources of energy were foods from the milk (27.4%), cereals (22.1%), and breads (14.1%)groups. Energy intake at breakfast was no different in children whose household income was at or above the low-income cutoff than in children whose household income was below the cutoff. While few children missed breakfast, many needed more energy at this meal, and non-economic as well as economic influences on breakfast consumption need to be identified. PMID- 12645547 TI - The hemoglobin system of Antarctic and non-Antarctic notothenioid fishes. AB - Studies of the hemoglobin system of fish of the suborder Notothenioidei have been extended to non-Antarctic species of Pseudaphritis urvillii and Notothenia angustata. The two species belong to families that were the first to diverge within the suborder. The degree of amino acid sequence identity with Antarctic notothenioids and other non-Antarctic fish species is analyzed with respect to phyletic and ecological diverence. PMID- 12645546 TI - A novel mitochondrial Ca2+-dependent solute carrier in the liver identified by mRNA differential display. AB - Pancreatic AR42J cells have the feature of pluripotency of the precursor cells of the gut endoderm. Dexamethasone converts them to exocrine cells or liver cells. Using mRNA differential display techniques, we have identified a novel Ca2+ dependent member of the mitochondrial solute carrier superfamily, which is expressed during the course of differentiation, and have designated it MCSC. The corresponding cDNA comprises an open reading frame of 1407 base pairs encoding a polypeptide of 469 amino acids. The carboxyl-terminal-half of MCSC has high similarity with other mitochondrial carriers, and the amino-terminal-half has three canonical elongation factor-hand motifs and has calcium binding capacity. The deduced amino acid sequence revealed 79.1% homology to the rabbit peroxisomal Ca2+-dependent member of the mitochondrial superfamily, but the subcellular localization of the protein was exclusively mitochondrial, not peroxisomal. Northern blot and Western blot analyses revealed its predominant expression in the liver and the skeletal muscle. In the liver, the expression level of MCSC was higher in the adult stage than in the fetal stage, and MCSC was highly up regulated in dexamethasone-treated AR42J cells before the expression of albumin. Taken together, MCSC may play an important role in regulating the function of hepatocytes rather than in differentiation in vivo. PMID- 12645548 TI - Atrial chamber-specific expression of sarcolipin is regulated during development and hypertrophic remodeling. AB - Intracellular Ca2+ regulation is critical in the normal cardiac function and development of pathologic hearts. Phospholamban, an endogenous inhibitor of sarcoplasmic reticulum Ca2+ ATPase in the sarcoplasmic reticulum, plays an important role in Ca2+ cycling in heart. Recently, sarcolipin has been identified as having a similar function as phospholamban in skeletal muscle. Because phospholamban is differentially expressed in atrial and ventricular myocardia and its expression is often altered in diseased hearts, we investigated the cardiac chamber specificity of sarcolipin expression and its regulation during development and hypertrophic remodeling. Northern blot analysis revealed that the expression of mouse sarcolipin mRNA was most abundant in the atria and was undetectable in the ventricles, indicating an atrial chamber-specific expression pattern. Atrial chamber-specific expression of sarcolipin mRNA was increased during development. These findings were confirmed by in situ hybridization studies. In addition, sarcolipin expression was down-regulated in the atria of hypertrophic heart when induced by ventricular specific overexpression of the activated H-ras gene. In humans, sarcolipin mRNA was also expressed in the atria but not detected in the ventricles, although sarcolipin expression was most abundant in skeletal muscle. Taken together, sarcolipin is likely to be an atrial chamber-specific regulator of Ca2+ cycling in heart. PMID- 12645549 TI - The gene patenting controversy: a convergence of law, economic interests, and ethics. PMID- 12645550 TI - What about my right to privacy? Where the Court went wrong in Ferguson v. City of Charleston. PMID- 12645551 TI - [What do you see? Subacute cutaneous lupus erythematosis (SCLE)]. PMID- 12645552 TI - Who owns my DNA?: the national and international intellectual property law on human embryonic tissue and cloning. PMID- 12645553 TI - Should a ban on reproductive cloning include a ban on cloning for purposes of research or therapy? PMID- 12645554 TI - The constitutional implications of a cloning society. PMID- 12645555 TI - Death wish: resuscitating self-determination for the critically ill. PMID- 12645557 TI - [Embracing the changing treatment modalities for cerebrovascular diseases]. PMID- 12645556 TI - What happens if Roe is overruled? Extraterritorial regulation of abortion by the states. PMID- 12645558 TI - Withdrawal of treatment for minors in a persistent vegetative state: parents should decide. PMID- 12645559 TI - [Current neurologists focus more on clinical practice]. PMID- 12645560 TI - Law and human genetics on the threshold of the new millennium. Introduction. PMID- 12645561 TI - The man on the moon, immortality, and other millennial myths: the prospects and perils of human genetic engineering. PMID- 12645562 TI - Re-examining the role of patents in appropriating the value of DNA sequences. PMID- 12645563 TI - Medical genetics: present and future benefits. PMID- 12645564 TI - The Human Genome Project and public perception: truth and consequences. PMID- 12645565 TI - Re-engineering the laws of organ transplantation. PMID- 12645566 TI - Enhancement of B-MYB transcriptional activity by ZPR9, a novel zinc finger protein. AB - By using the yeast two-hybrid system, the zinc finger protein ZPR9 was identified as one of the B-MYB interacting proteins that associates with the carboxyl terminal conserved region of B-MYB. ZPR9 was found to form in vivo complexes with B-MYB, as demonstrated by in vivo binding assay and coimmunoprecipitation experiments of the endogenously and exogenously expressed proteins. Deletion analysis revealed that this binding was mediated by all three functional domains, an amino-terminal DNA-binding domain, a transactivation domain, and a carboxyl terminal conserved region of B-MYB. We show that the interaction of ZPR9 with B MYB is functional because cotransfection of ZPR9 significantly up-regulates B-MYB transcriptional activity in a dose-dependent manner. In addition, coexpression of ZPR9 with B-MYB caused the accumulation of B-MYB, as well as ZPR9, in the nucleus. Furthermore, constitutive expression of ZPR9 in human neuroblastoma cells induces apoptosis in the presence of retinoic acid. These results strongly suggest that ZPR9 plays an important role in modulation of the transactivation by B-MYB and cellular growth of neuroblastoma cells. PMID- 12645567 TI - The herpes simplex virus type-1 single-strand DNA-binding protein (ICP8) promotes strand invasion. AB - ICP8, the herpes simplex virus type-1 single-strand DNA-binding protein, was recently shown to promote strand exchange in conjunction with the viral replicative helicase (Nimonkar, A. V., and Boehmer, P. E. (2002) J. Biol. Chem. 277, 15182-15189). Here we show that ICP8 also catalyzes strand invasion in an ATP-independent manner. Thus, ICP8 promotes the assimilation of a single-stranded donor molecule into a homologous plasmid, resulting in the formation of a displacement loop. Invasion of a homologous duplex by single-stranded DNA requires homology at either 3' or 5' end of the invading strand. The reaction is dependent on the free energy of supercoiling and alters the topology of the acceptor plasmid. Hence, strand invasion products formed by ICP8 are resistant to the action of restriction endonucleases that cleave outside of the area of pairing. The ability to catalyze strand invasion is a novel activity of ICP8 and the first demonstration of a eukaryotic viral single-strand DNA-binding protein to promote this reaction. In this regard ICP8 is functionally similar to the prototypical prokaryotic recombinase RecA and its eukaryotic homologs. This strand invasion activity of ICP8 coupled with DNA synthesis may explain the high prevalence of branched DNA structures during viral replication. PMID- 12645568 TI - Health and the economy: empowerment through evidence. PMID- 12645569 TI - The cases of John Bunyan. Part 2. James and Janet. PMID- 12645570 TI - The British reaction to dementia praecox 1893-1913. Part 2. AB - Part 1 of this study described the backdrop to the development of Kraepelin's ideas on dementia praecox and examined the response to the concept in the British psychiatric textbooks and journals of the period. Part 2 now explores the reaction to the concept in the professional meetings of the period, held by the British Medical Association (BMA) and the Medico-Psychological Association (MPA) during the years 1893 to 1913. In addition, it examines and evaluates the main issues and conclusions arising from the debate. PMID- 12645572 TI - The distinction between witchcraft and madness in colonial Connecticut. AB - This essay argues two points in regard to early New England: first, that witchcraft is not a significant aspect of the history of mental illness; and second, that seventeenth-century society had a cultural protocol for distinguishing one from the other. The examples discussed in detail are from Connecticut, but they are representative of colonial New England as a whole. PMID- 12645571 TI - Catalytic properties of mutant 23 S ribosomes resistant to oxazolidinones. AB - Kinetic analysis of ribosomal peptidyltransferase activity in a methanolic puromycin reaction with wild type and drug-resistant 23 S RNA mutants was used to probe the structural basis of catalysis and mechanism of resistance to antibiotics. 23 S RNA mutants G2032A and G2447A are resistant to oxazolidinones both in vitro and in vivo with the latter displaying a 5-fold increase in the value of Km for initiator tRNA and a 100-fold decrease in Vmax in puromycin reaction. Comparison of the Ki values for oxazolidinones, chloramphenicol, and sparsomycin revealed partial cross-resistance between oxazolidinones and chloramphenicol; no cross-resistance was observed with sparsomycin, a known inhibitor of the peptidyltransferase A-site. Inhibition of the mutants using a truncated CCA-Phe-X-Biotin fragment as a P-site substrate is similar to that observed with the intact initiator tRNA, indicating that the inhibition is substrate-independent and that the peptidyltransferase itself is the oxazolidinone target. Mapping of all known mutations that confer resistance to these drugs onto the spatial structure of the 50 S ribosomal subunit allows for docking of an oxazolidinone into a proposed binding pocket. The model suggests that oxazolidinones bind between the P- and A-loops, partially overlapping with the peptidyltransferase P-site. Thus, kinetic, mutagenesis, and structural data suggest that oxazolidinones interfere with initiator fMet-tRNA binding to the P site of the ribosomal peptidyltransferase center. PMID- 12645573 TI - Development and termination of the working relationship of C. G. Jung and Eugen Bleuler 1900-1909. AB - From 1900 to 1909, C. G. Jung was on the staff of the University Psychiatric Clinic in Zurich ('Burgholzli'), headed by Eugen Bleuler, where he became a senior physician (Sekundararzt) in 1905. Jung received his doctorate (1902) and postdoctoral qualification as a university lecturer (Habilitation) based on Bleuler's reports to the department of medicine, and he continued to give lectures on certain recurring topics, such as the theory of hysteria, until summer semester 1914, well after his resignation. Minutes of department meetings that have not previously been evaluated appear to indicate that a major factor leading to Jung's resignation on 7 March 1909 was the department's rejection of the founding of the 'psychopathologic research laboratory' that he had expected. The importance of other events for Bleuler's relationship with Jung - the unsuccessful attempt at an analytical treatment of Otto Gross, and Jung's relationship with Sabina Spielrein - cannot be determined from current sources. PMID- 12645574 TI - A blocking antibody to the hyaluronan receptor for endocytosis (HARE) inhibits hyaluronan clearance by perfused liver. AB - Hyaluronan (HA) and chondroitin sulfate clearance from lymph and blood is mediated by the hyaluronan receptor for endocytosis (HARE). The purification and molecular cloning (Zhou, B., Weigel, J. A., Saxena, A., and Weigel, P. H. (2002) Mol. Biol. Cell 13, 2853-2868) of this cell surface receptor were finally achieved after we developed monoclonal antibodies (mAbs) against HARE. There are actually two independent isoreceptors for HA, which in rat are designated the 175 kDa HARE and 300-kDa HARE. Only one mAb (number 174) effectively and completely blocked the specific uptake of 125I-HA at 37 degrees C by rat liver sinusoidal endothelial cells. 125I-HA binding to both the 175-kDa and 300-kDa HARE proteins in a ligand blot assay was almost completely inhibited by <1 microg/ml mAb-174, whereas mouse IgG had little or no effect. MAb-174 also performed very well in Western analysis, indirect fluorescence microscopy, and a variety of immuno procedures. Immunohistochemistry using mAb-174 localized HARE to the sinusoidal cells of rat liver, spleen, and lymph node. Western analysis using mAb-174 revealed that the sizes of both HARE glycoproteins were the same in these three tissues. 125I-HA was taken up and degraded by excised rat livers that were continuously perfused ex vivo with a recirculating medium. This HA clearance and metabolism by liver, which is a physiological function of HARE, was very effectively blocked by mAb-174 but not by mouse IgG. The results indicate that mAb-174 will be a useful tool to study the functions of HARE and the physiological significance of HA clearance. PMID- 12645575 TI - Inducible expression of a dominant negative DNA polymerase-gamma depletes mitochondrial DNA and produces a rho0 phenotype. AB - We report the inducible, stable expression of a dominant negative form of mitochondria-specific DNA polymerase-gamma to eliminate mitochondrial DNA (mtDNA) from human cells in culture. HEK293 cells were transfected with a plasmid encoding inactive DNA polymerase-gamma harboring a D1135A substitution (POLGdn). The cells rapidly lost mtDNA (t1/2 = 2-3 days) when expression of the transgene was induced. Concurrent reduction of mitochondrial encoded mRNA and protein, decreased cellular growth rate, and compromised respiration and mitochondrial membrane potential were observed. mtDNA depletion was reversible, as demonstrated by restoration of mtDNA copy number to normal within 10 days when the expression of POLGdn was suppressed following a 3-day induction period. Long term (20 days) expression of POLGdn completely eliminated mtDNA from the cells, resulting in rho0 cells that were respiration-deficient, lacked electron transport complex activities, and were auxotrophic for pyruvate and uridine. Fusion of the rho0 cells with human platelets yielded clonal cybrid cell lines that were populated exclusively with donor-derived mtDNA. Respiratory function, mitochondrial membrane potential, and electron transport activities were restored to normal in the cybrid cells. Inducible expression of a dominant negative DNA polymerase gamma can yield mtDNA-deficient cell lines, which can be used to study the impact of specific mtDNA mutations on cellular physiology, and to investigate mitochondrial genome function and regulation. PMID- 12645576 TI - Response to Amar J. Klar: The chromosome 1;11 translocation provides the best evidence supporting genetic etiology for schizophrenia and bipolar affective disorders. PMID- 12645578 TI - Correct catheter connections. PMID- 12645577 TI - c-Src-dependent tyrosine phosphorylation of IKKbeta is involved in tumor necrosis factor-alpha-induced intercellular adhesion molecule-1 expression. AB - The signaling pathway involved in tumor necrosis factor-alpha (TNF-alpha)-induced intercellular adhesion molecule-1 (ICAM-1) expression was further studied in human A549 epithelial cells. TNF-alpha- or 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ICAM-1 promoter activity was inhibited by a protein kinase C (PKC) inhibitor (staurosporine), tyrosine kinase inhibitors (genistein and herbimycin A), or an Src-specific tyrosine kinase inhibitor (PP2). TNF-alpha- or TPA-induced IkappaBalpha kinase (IKK) activation was also blocked by these inhibitors, which slightly reversed TNF-alpha-induced but completely reversed TPA-induced IkappaBalpha degradation. c-Src and Lyn, two members of the Src kinase family, were abundantly expressed in A549 cells, and their activation by TNF-alpha or TPA was inhibited by the same inhibitors. Furthermore, the dominant-negative c-Src (KM) mutant inhibited induction of ICAM-1 promoter activity by TNF-alpha or TPA. Overexpression of the constitutively active PKC or wild-type c-Src plasmids induced ICAM-1 promoter activity, this effect being inhibited by the dominant negative c-Src (KM) or IKKbeta (KM) mutant but not by the nuclear factor-kappaB inducing kinase (NIK) (KA) mutant. The c-Src (KM) mutant failed to block induction of ICAM-1 promoter activity caused by overexpression of wild-type NIK. In co-immunoprecipitation and immunoblot experiments, IKK was found to be associated with c-Src and to be phosphorylated on tyrosine residues after TNF alpha or TPA treatment. Two tyrosine residues, Tyr188 and Tyr199, near the activation loop of IKKbeta, were identified as being important for NF-kappaB activation. Substitution of these residues with phenylalanines abolished ICAM-1 promoter activity and c-Src-dependent phosphorylation of IKKbeta induced by TNF alpha or TPA. These data suggest that, in addition to activating NIK, TNF-alpha also activates PKC-dependent c-Src. These two pathways converge at IKKbeta and go on to activate NF-kappaB, via serine phosphorylation and degradation of IkappaB alpha, and, finally, to initiate ICAM-1 expression. PMID- 12645579 TI - Clarification. PMID- 12645580 TI - Performing peritoneal dialysis. PMID- 12645581 TI - Hold tight: keeping catheters secure. PMID- 12645582 TI - Recognizing normeperidine toxicity. PMID- 12645583 TI - Identification of glycosphingolipid receptors for pierisin-1, a guanine-specific ADP-ribosylating toxin from the cabbage butterfly. AB - Pierisin-1, a cytotoxic protein found naturally in the cabbage butterfly, induces apoptosis of mammalian cells. Our recent studies suggest that pierisin-1 consists of an N-terminal ADP-ribosyltransferase domain, and a C-terminal region that binds to receptors on the surfaces of target cells and incorporates the protein into cells. The present study was undertaken to identify receptors for pierisin 1. The cross-linking and cloning experiments suggested that the proteins on cell membrane had no binding ability to pierisin-1. Inhibitory assays of fractionated lipids from human cervical carcinoma HeLa cells, which are highly sensitive to pierisin-1, indicated neutral glycosphingolipids on the cell surface to show receptor activity. Inhibitory assays and TLC immunostaining using anti-pierisin-1 antibodies demonstrated two neutral glycosphingolipids as active components. Analysis of their structures with glycosphingolipid-specific antibodies and negative secondary ion mass spectrometry identified them as globotriaosylceramide (Gb3) and globotetraosylceramide (Gb4). The receptor activities of Gb3 and Gb4 for pierisin-1 were also confirmed with these authentic compounds. Pierisin-1 insensitive mouse melanoma MEB4 cells were found to lack pierisin-1 receptors, including Gb3 and Gb4, but pretreatment of the cells with glycosphingolipid Gb3 or Gb4 enhanced their sensitivity to pierisin-1. Thus, Gb3 and Gb4 were proven to serve as pierisin-1 receptors. The C-terminal region of pierisin-1 consists of possible lectin domains of a ricin B-chain, containing QXW sequences, which are essential for its structural organization. Alteration of QXW by site-directed mutagenesis caused marked reduction of pierisin-1 cytotoxicity. Thus, our results suggest that pierisin-1 binds to Gb3 and Gb4 receptors at the C-terminal region, in a manner similar to ricin, and then exhibits cytotoxicity after incorporation into the cell. PMID- 12645584 TI - A normality bias in legal decision making. AB - It is important to understand how legal fact finders determine causation and assign blame. However, this process is poorly understood. Among the psychological factors that affect decision makers are an omission bias (a tendency to blame actions more than inactions [omissions] for bad results), and a normality bias (a tendency to react more strongly to bad outcomes that spring from abnormal rather than normal circumstances). The omission and normality biases often reinforce one another when inaction preserves the normal state and when action creates an abnormal state. But what happens when these biases push in opposite directions as they would when inaction promotes an abnormal state or when action promotes a normal state? Which bias exerts the stronger influence on the judgments and behaviors of legal decision makers? The authors address this issue in two controlled experiments. One experiment involves medical malpractice and the other involves stockbroker negligence. They find that jurors pay much more attention to the normality of conditions than to whether those conditions arose through acts or omissions. Defendants who followed a nontraditional medical treatment regime or who chose a nontraditional stock portfolio received more blame and more punishment for bad outcomes than did defendants who obtained equally poor results after recommending a traditional medical regime or a traditional stock portfolio. Whether these recommendations entailed an action or an omission was essentially irrelevant. The Article concludes with a discussion of the implications of a robust normality bias for American jurisprudence. PMID- 12645585 TI - Using a colonoscopy to survey intestinal health. PMID- 12645587 TI - How do I manage a lumbar drain? PMID- 12645586 TI - Using human skin equivalents to heal chronic wounds. PMID- 12645588 TI - Check that cable! PMID- 12645589 TI - [The strategy thinking on the research and development in the technology of stomatology with our own intellectual property rights]. PMID- 12645590 TI - [Evaluation of therapeutic concept and methods for infected pulp]. PMID- 12645592 TI - At the Crichton Royal with William Mayer-Gross (b. 15 Jan. 1889, d. 15 Feb. 1961). PMID- 12645591 TI - Ewald Hecker (1843-1909). PMID- 12645593 TI - Can research subjects of clinical trials in developing countries sue physician investigators for human rights violations? PMID- 12645594 TI - Acknowledging the hypocrisy: granting minors the right to choose their medical treatment. PMID- 12645595 TI - Informed consent: does the First Amendment protect a patient's right to choose alternative treatment? PMID- 12645596 TI - The name game: issues surrounding New York state's HIV partner notification law. PMID- 12645597 TI - The hunting of the snark: the moral status of embryos, right-to-lifers, and Third World women. PMID- 12645599 TI - The legacy of Kraepelin (essay review). PMID- 12645598 TI - Commodification and commercialization in human embryo research. PMID- 12645600 TI - Policy issues in donor insemination. PMID- 12645601 TI - Hidden treasures in the history of psychiatry (review of dissertations). PMID- 12645602 TI - Infertility as a public health problem: why assisted reproductive technologies are not the answer. PMID- 12645604 TI - Solving the physician's dilemma: an HIV partner-notification plan. PMID- 12645603 TI - Financing high-tech reproductive medical expenditures. PMID- 12645605 TI - Women and HIV: a gender-based analysis of a disease and its legal regulation. PMID- 12645606 TI - The Americans with Disabilities Act and the reproductive rights of HIV-infected women. PMID- 12645607 TI - Change of venue: abortion regulation in the states. PMID- 12645608 TI - Equality theory and reproductive freedom. PMID- 12645609 TI - Bibliography of congenital muscular dystrophies: Series V (2001). PMID- 12645610 TI - What's so bad about human cloning? AB - There appears to be a consensus in the general community that reproductive cloning is an immoral technology that should be banned. It may, however, be argued, at least from the perspective of the Jewish tradition, that reproductive cloning has many positive benefits. It is thus essential that one carefully weigh the costs and the benefits before deciding on a definitive course of action. PMID- 12645611 TI - Revisiting the definition of homo sapiens. AB - Research in genomics, human cloning, and transgenic technology has challenged bioethicists and scientists to rethink the definition of human beings as a species. For example, should the definition incorporate a genetic criterion and how does the capacity to genetically engineer human beings affect the definition of our species? In considering these contemporary bioethical dilemmas, we revisit an ancient source, the Talmud, and highlight how it provides specific biological, cultural, and genetic criteria to define the human species. PMID- 12645612 TI - Informed consent and research involving the newly dead. AB - This paper examines informed consent in relation to research involving the newly dead. Reasons are presented for facilitating advance decision making in relation to postmortem research, and it is argued that the informed consent of family members should be sought when the deceased have not made a premortem decision. Regardless of whether the dead can be harmed, there are two important respects in which family consent can serve to protect the dead: (1) protecting the deceased's body from being used for research that is incompatible with the person's postmortem preferences and values and (2) protecting the deceased's body's from being subject to disrespectful treatment. These claims are explained and justified, and several objections are critically examined. Additional reasons for securing family consent are presented including to protect them from additional emotional distress, to respect their wishes about wanting to have a say, and to maintain public trust in the medical profession and medical research. The paper also examines the scope of disclosure in relation to postmortem research. PMID- 12645614 TI - IOM report on the system for protecting human research participants. PMID- 12645613 TI - Beyond the medical model? Disability, formal justice, and the exception for the "profoundly impaired". AB - The formal justice model proposed by Anita Silvers in Disability, Discrimination, and Difference emphasizes the social model of disability and the need for full equality of opportunity, and it suggests that a distributive model of justice that gives special benefits to individuals with disabilities is self-defeating. Yet in that work, Silvers allows an exception for the "profoundly impaired." In this paper, I show how the formal justice theory falls short when it comes to defining and dealing with "profoundly impaired" individuals and explore the ways in which making the exception raises serious theoretical concerns for the grounding of the formal justice model. PMID- 12645615 TI - Bioethics and cloning, part II. PMID- 12645616 TI - Volunteer dentistry in Guatemala. PMID- 12645617 TI - Application of a novel protein biochip technology for detection and identification of rheumatoid arthritis biomarkers in synovial fluid. AB - We compared protein profiles of the synovial fluid of patients with rheumatoid arthritis and osteoarthritis by using surface-enhanced laser desorption/ionization mass spectrometry technology. With this approach, we identified a protein expressed specifically in the synovial fluid of the patients with rheumatoid arthritis. During the investigation, we found several reproducible and discriminatory biomarker candidates for distinction between rheumatoid arthritis and osteoarthritis. Among these candidates, a 10 850 Da protein peak was the clearest example of a single signal found specifically in the rheumatoid arthritis samples. This candidate was purified using a size exclusion spin column followed by gel electrophoresis and subsequently identified by peptide mapping and post-source decay (PSD) analysis. The results clearly indicate that the protein is myeloid-related protein 8, which was verified by the enzyme immunoassay. It is known that the myeloid-related protein 8 level in serum and synovial fluid is related to disease activity in juvenile rheumatoid arthritis. The results suggest that the ProteinChip platform is useful to detect and identify protein biomarkers expressed specifically in diseases or in some stage of diseases. PMID- 12645618 TI - Analysis of the adenovirus type 5 proteome by liquid chromatography and tandem mass spectrometry methods. AB - We compared detection sensitivity and protein sequence coverage of the adenovirus type 5 proteome achievable by liquid chromatography and tandem mass spectroscopy (LC/MS/MS) using three sample preparation and clean up methods. Tryptic digestion was performed on either purified viral proteins or whole virus, and followed by shotgun sequencing using tandem mass spectrometry for peptide identification. We used a recombinant adenovirus type 5 as a test system. The methods included separation of adenoviral proteins by reversed-phase high-performance liquid chromatography followed by tryptic digestion and analysis by LC/MS/MS. Alternatively, the purified whole virus was digested with trypsin and the peptides separated either by one-dimensional (reversed-phase) or by two dimensional (cation exchange and reversed-phase) chromatography and analyzed by tandem mass spectrometry. A total of 11 protein species were identified from 154 peptides. All of the major viral proteins were found. In addition, two minor proteins, the 23 kDa viral protease and the late L1 protein, were identified for the first time by chromatography based assays. The 23 kDa viral protease, present at only 10 copies per virus, and representing 0.2% of the protein content of the virus, was detected by the 2D LC/MS/MS analysis of the whole virus digest from a sample containing only 70 fmols of the protein. This demonstrates the high sensitivity and selectivity of the method. The 2D LC/MS/MS analysis of the whole virus digest was also able to detect all viral proteins with copy numbers at or above 10/virus particle, with broad coverage of the amino acid sequences. Coverage ranged from 2 to 54%, a majority between 20 and 35%, suggesting the possibility of using this analysis to assess the purity of the virus preparations. This broad coverage may also provide a useful approach to identify posttranslational modifications on the structural proteins of the adenovirus. PMID- 12645619 TI - Foldability and the amino acid compositions of exons and introns. AB - Various procedures are employed to relate the structural tendencies of polypeptide chain fragments to amino acid residues that in general have low background frequencies. A numerical evaluation of the content of these amino acids, named amino acid diversity, is defined. Distributions of the amino acid diversity parameter in databases containing exons, introns, and randomized exons show that there is a small difference between exons and shuffled exons, a detectable difference between exons and introns, and a large difference between exons and totally randomized exons. PMID- 12645620 TI - Sequential analysis of N- and O-linked glycosylation of 2D-PAGE separated glycoproteins. AB - A robust method has been developed that allows analysis of both N- and O-linked oligosaccharides released from glycoproteins separated using 2D-PAGE and then electroblotted to PVDF membrane. This analysis provides efficient oligosaccharide profiling applicable to glycoproteomic analysis. The method involves the enzymatic release of N-linked oligosaccharides using PNGase F followed by the chemical release of O-linked oligosaccharides using reductive beta-elimination and analysis using LC-ESI-MS. Oligosaccharides from the major plasma glycoproteins with a pI between 4 and 7 were characterized from the glycoforms of haptoglobin, alpha2-HS-glycoprotein, serotransferrin, alpha1-antitrypsin, and alpha1-antichymotrypsin. It was shown that the separation of protein glycoforms evident in 2D-PAGE is partially due to the combined sialylation of the O-linked and N-linked oligosaccharides. Bi-, tri- and tetra-antennary N-linked structures, which had differing levels of sialylation and fucosylation, were found to be present on the glycoproteins analyzed, together with O-linked oligosaccharides such as mono-, and disialylated T-antigen and a disialylated core type 2 hexasaccharide. In addition, N-linked site-specific information was obtained by MALDI-MS analysis using tryptic digestion after PNGase F release of the oligosaccharides. PMID- 12645621 TI - High-throughput proteomics: a flexible and efficient pipeline for protein production. AB - Many studies that aim to characterize the proteome require the production of pure protein in a high-throughput format. We have developed a system for high throughput subcloning, protein expression and purification that is simple, fast, and inexpensive. We utilized ligation-independent cloning with a custom-designed vector and developed an expression screen to test multiple parameters for optimal protein production in E. coli. A 96-well format purification protocol that produced microgram quantities of pure protein was also developed. PMID- 12645622 TI - Microcolumn capture and digestion of proteins combined with mass spectrometry for protein identification. AB - A procedure has been developed for protein identification using mass spectrometry (MS) that incorporates sample cleanup, preconcentration, and protein digestion in a single-stage system. The procedure involves the adsorption of a protein, or protein mixture, from solution onto a hydrophobic resin that is contained within a microcolumn. Sample loading is accomplished by flowing the protein solution through the microcolumn, where the protein adsorbs to the hydrophobic surface. The protein is digested while still bound to the hydrophobic surface by flowing a buffered trypsin solution through the column bed. The peptide fragments are subsequently eluted for detection by MALDI or ESI-MS. The procedure is demonstrated using dilute protein samples containing high concentrations of salt, urea, and modest amount of sodium dodecyl sulfate relative to protein. Peptide fragments are also detected by MS from a 500 nM bacteriorhodopsin solution digested in a microcolumn. In this case, a combined cyanogen bromide/trypsin digestion was performed in-column. The procedure is applied to the MALDI-MS/MS identification of proteins present in an individual fraction collected by ion exchange HPLC separation of E. coli total cell extract. An additional application is illustrated in the analysis of a human plasma fraction. A total of 14 proteins, which were present in the sample at sub-micromolar concentrations, were identified from ESI-MS/MS. The microcolumn digestion procedure represents the next step toward a system for fully automated protein analysis through capture and digestion of the adsorbed protein on hydrophobic surfaces. PMID- 12645623 TI - Dissociations of disulfide-linked gaseous polypeptide/protein anions: ion chemistry with implications for protein identification and characterization. AB - Ion trap collisional activation of whole protein anions that contain disulfide bonds results in the cleavage of one of the bonds that comprises the disulfide linkage. The disulfide linkage can break at any of three possible locations, giving rise to several products with different partitioning of sulfur atoms. A facile second-generation dissociation occurs at the polypeptide backbone from products formed from cleavage of the nearest C-S bond of a disulfide linkage. This cleavage occurs exclusively at the N-terminal side of the cysteine residue, from which the C-S bond was cleaved, thereby yielding c and z-S type product ions. This secondary reaction is apparently a relatively low-energy reaction with relatively high entropy requirements because it is not observed to be a major process under beam-type collisional activation conditions, but is a major process under ion trap collisional activation conditions. The specificity of this cleavage, as well as the ability to distinguish it from other cleavages by the sulfur atom distribution, make it useful for the identification of unknown proteins via database searching. Furthermore, the pattern of disulfide cleavages can be useful in providing information about the location of post-translational modifications. Examples using bovine pancreatic trypsin inhibitor and ribonuclease A and B are given to illustrate these points. PMID- 12645625 TI - High-throughput peptide mass mapping using a microdevice containing trypsin immobilized on a porous polymer monolith coupled to MALDI TOF and ESI TOF mass spectrometers. AB - An enzymatic microreactor with a volume of 470 nL has been prepared by immobilizing trypsin on a 10 cm long reactive porous polymer monolith located in a 100 microm i.d. fused silica capillary. This reactor affords suitable degrees of digestion of proteins even after very short residence times of less than 1 min. The performance is demonstrated with the digestion of eight proteins ranging in molecular mass from 2848 to 77 754. The digests were analyzed using mass spectrometry in two modes: off-line MALDI and in-line nanoelectrospray ionization. The large numbers of identified peptides enable a high degree of sequence coverage and positive identification of the proteins. The extent of sequence coverage decreases as the molecular mass of the digested protein increases. PMID- 12645624 TI - HeLa cell adhesion on various collagen-grafted surfaces. AB - Cell adhesion is important to develop cell microarrays and biocompatible materials. Collagen has been reported to be able to improve cell adhesion. In this paper, two collagen coating methods (collagen grafted directly on the substrate and chitosan-modified substrate) were carried out, on which the adhesive behaviors of HeLa cells were studied. An atomic force microscope and a surface potential meter were used to characterize morphologies and electric polarization of these surfaces. It was found that surface electric polarization and the its durability and surface topography were key factors to cell adhesion. Collagen (1 mg/mL) grafted on 1% chitosan-modified surface showed the best adhesion of HeLa cell. This work might be helpful to the practical application of cell microarray chips. PMID- 12645626 TI - Kinetics of mononuclear cell infiltration and cytokine expression in iodine induced thyroiditis in the NOD-H2h4 mouse. AB - Mononuclear cell infiltration of the thyroid gland is a common histologic feature of chronic lymphocytic thyroiditis. Although the infiltrating mononuclear cells have been implicated in the destruction of the thyroid, information concerning the progression of infiltration into the thyroid is limited. In this report, we examine the composition and kinetics of mononuclear cell infiltration in the thyroid and the expression of major histocompatibility complex class II (I-Ak), IL-12, and IFN-gamma in the thyroid of the NOD-H2h4 mouse, a model of spontaneous autoimmune thyroiditis accelerated by the administration of excess dietary iodine. Mice were given a low dose of 0.015% NaI in their drinking water for 2, 4, 6, 8, and 16 weeks, and thyroids were removed, serially sectioned, and stained in an avidin-biotin peroxidase assay. The thyroid infiltrate included CD4+ and CD8+ T cells, F4/80+ macrophages, and B220+ B cells. After 2 weeks of iodine treatment, CD4+ T cells were the first seen in the thyroid, followed by CD8+ T cells and F4/80+ macrophages. B220+ B cells entered the thyroid after 4 weeks of iodine treatment. IL-12 and IFN-gamma positive cells were located in the thyroid early in disease and were up-regulated in the focal accumulations of infiltrating cells. Thyrocytes clearly expressed I-Ak after 4 weeks of iodine treatment near the location of mononuclear cell infiltration. PMID- 12645627 TI - Expression of granzyme B mRNA is altered in human immunodeficiency virus infected patients. AB - CD8-positive cytotoxic T lymphocytes and natural killer cells are the major cytotoxic components of the antiviral immune response. The major pathway used by these cells in response to viral-infected cells involves granzymes, cytotoxic granule serine proteases involved in the pathway leading to target cell DNA fragmentation and apoptosis. The levels of granzyme B mRNA in peripheral blood cells of human immunodeficiency virus (HIV) type-1 infected patients in comparison to noninfected individuals were assessed by quantitative competitive reverse transcriptase polymerase chain reaction. Expression of granzyme B mRNA is altered in HIV-1 infected patients. Significantly fewer HIV patients had detectable granzyme B mRNA levels than controls. The one HIV-infected patient with detectable granzyme B mRNA displayed a much higher level of this mRNA than all healthy controls. Cell-mediated cytotoxicity during HIV-1 infection may be impaired due to a deficient quantity of active cytotoxic granules or to their abnormal regulation. PMID- 12645629 TI - Different influences of hyperglycemic duration on phosphorylated extracellular signal-regulated kinase 1/2 in rat heart. AB - Extracellular signal-regulated kinase (ERK) 1/2 is an important intracellular proteinase associated with myocardial protection against heart injury. Hyperglycemia was also reported to be highly involved in heart injury by the formation of advanced glycation end products (AGEs) in myocardial protein, resulting in its altered structure and function. However, the effect of this glycation on mitogen-activated protein kinases, particularly ERK1/2, in the myocardium is largely unclarified. In this study, we investigated whether the glycation of an intracellular protein, ERK1/2, would result in ERK1/2-AGEs formation that adversely affects ERK1/2 activation in the rat heart under hyperglycemia. Hyperglycemia was induced by injection of streptozotocin (STZ) and hearts were examined 4 and 20 weeks after STZ treatment. By immunohistochemical staining and Western blotting, it was determined that the level of phosphorylated ERK1/2 in the rat heart under hyperglycemia 20 weeks after STZ treatment decreased markedly by about 50% of that of the time-matched control group, whereas in the case of 4 weeks after STZ treatment, it increased by about 2.7 fold that of the time-matched group. The level of deposition of AGEs in proteins of the myocardium increased significantly depending on the duration of hyperglycemia. Twenty weeks after STZ treatment, two clear bands corresponding to 44- and 42-kDa AGEs were detected by Western blotting: these corresponded to protein sizes of ERK1/2. The immunoprecipitation method further confirmed the formation and the increased intensity of ERK1/2-AGEs in the rat heart under hyperglycemia for 20 weeks. These results demonstrate that long-term hyperglycemia may inhibit ERK1/2 phosphorylation in the myocardium, whereas a short-term (4 weeks) hyperglycemia enhances its phosphorylation. The ERK1/2 phosphorylation under long-term hyperglycemia is very different from that under short-term hyperglycemia. In addition, this inhibition of ERK1/2 activation appears to be dependent on the formation of ERK1/2-AGEs under long-term hyperglycemia, which may be related in part to the etiology of diabetic cardiomyopathy. It also suggests that the formation of AGEs in intracellular enzymes and proteins under hyperglycemia could play important roles in the development of diabetes complications. PMID- 12645628 TI - Neuro-Behcet's disease: a masquerader of multiple sclerosis. A prospective study of neurologic manifestations of Behcet's disease in 96 Iranian patients. AB - Behcet's disease is a multisystem vasculitis. Its neurologic complications include different syndromes. The purpose of this investigation was to study the prevalence of neurologic manifestations among patients with Behcet's disease and to determine the frequency of different symptoms, signs, and syndromes in neuro Behcet's disease. Ninety-six consecutive patients who were referred to the Behcet's Disease Clinic in Shiraz (southern Iran) were interviewed and thoroughly examined. Psychiatric evaluation, CSF analysis, electroencephalography, electrodiagnostic studies, and neuroradiologic imaging (preferably MRI) were performed in appropriate cases. Six patients (6.3%) had definite neuro-Behcet's disease. They were 4 males and 2 females (mean age 37.5 years). In 2 patients Behcet's disease had not been diagnosed before. The most frequent symptoms of neuro-Behcet's disease were headache (83.3%), paresthesia (83.3%), unsteadiness (66.7%), diplopia (66.7%), and weakness (50%). The most frequent signs were gait abnormalities (66.7%), sensory abnormalities (66.7%), ophthalmoplegia (50%), cerebellar ataxia (50%), and hemiplegia (50%). The most common syndrome was brain stem+ type (50%). Subacute onset and relapsing-remitting course were the most common temporal patterns. Neurological manifestation is a relatively less frequent complication of Behcet's disease but it produces severe disabilities. It must be considered in differential diagnosis of multiple sclerosis. PMID- 12645630 TI - Transient and ectopic expression of lumican by acinar cells in L-arginine-induced acute pancreatitis. AB - Lumican is a member of a small leucine-rich proteoglycan family. We previously found that lumican mRNA and its protein were ectopically and highly expressed in acinar cells in chronic pancreatitis (CP)-like lesions close to pancreatic cancer cells. CP-like lesions are characterized by acinar and ductal-ductular cell proliferation with expanding fibrosis. This finding suggests that lumican is ectopically synthesized by acinar cells under chronic inflammatory conditions and plays a role in fibrosis of the pancreas. However, the expression and role of lumican in acute inflammatory changes of the pancreas are not completely elucidated. In the present study, we aim to clarify whether lumican mRNA and its protein are expressed in exocrine or endocrine components in acute pancreatitis (AP). For experimental AP, Wistar rats received an intraperitoneal injection of L arginine. Western blot analysis showed an intense 50-kDa band corresponding to the lumican protein in normal and L-arginine-treated rat pancreas. After L arginine injection, three intense bands at 42, 57, and 92 kDa were detected on day 1. Immunohistochemically, the lumican protein was localized in ductal and a few centroacinar cells in the normal pancreas. After L-arginine injection, an immature fibrosis with fragmented and loose collagen fibers was observed in AP on day 4 and lumican immunoreactivity was detected in the collagen fibers. Lumican mRNA was faintly detected in islet cells in the normal pancreas, but it was strongly expressed in acinar and islet cells on day 1. Furthermore, lumican mRNA was expressed in many proliferating fibroblasts on day 4 by in situ hybridization. These findings indicate that lumican is transiently synthesized by acinar cells and fibroblasts in AP. Lumican proteins synthesized by acinar cells, islet cells, and fibroblasts may contribute to immature and transient fibrosis of AP. PMID- 12645631 TI - The short-lived exostosis induced surgically versus the lasting genetic hereditary multiple exostoses. AB - Hereditary osteochondromas are often caused by mutation in the EXT1 gene. The lesions are typified by formation of a "pseudo" growth plate like lesion growing at 60 degrees to the normal growth direction of the bone. Such lesions can be mimicked surgically by reverting the position--the polarity of the zone of LaCroix. The current study attempts to compare the pathology between EXT1 gene expression in humans and surgically created osteochondroma pathology in a rat model. Tissues of human bunion, human embryonal tissue, and human adult cartilage as well as normal rat epiphyses served as controls. Rats were operated on and a 60 degree span of the ring of LaCroix was inverted as described by Delgado (Delgado, E., Rodriguez, J. I., Serada, A., Tellez, M., and Pariagoa, R.. Clin. Orthop. 201, 251-258 (1985)). The surgically created osteochondromas were assessed by histology, histochemistry, and immunohistochemistry. The findings show that the surgically created lesions contain only a small amount of FGF receptor 3 (FGFR3) expressed on mesenchymal stem cells located in the perichondrium, as compared to the cell population carrying FGFR3 in the contralateral limb. Indian hedgehog and Bcl2 are downregulated, while BMP-2 is overexpressed in the operated limb, compared to the LaCroix ring of the contralaetral limb. The shortage, as well as the disturbed migration routes of the residual mesenchymal stem cells in surgically created osteochondromas leads eventually to resorption of the pathological elements. In search of additional markers characterizing such pathological structures composed of mesenchymal stem cells and cartilaginous and bony cells, EXT1 gene was found to be expressed in the surgically created osteochondromas, like in normal growth plates. Nitric oxide synthase was also expressed like in adult cartilage, though tumor necrosis factor alpha typifying Bunion formation was absent. In summary, surgically created osteochondromas lack the massive and continuous population of mesenchymal stem cells with Bcl2 expression. However, the small residual mesenchymal cell population gives rise to short-lived EXT1-expressing cells that disappear eventually due to spontaneous resorption. PMID- 12645632 TI - Pathogenesis of idiopathic scoliosis revisited. AB - Scoliosis, a lateral deviation of the spine frequently associated with rotation, is not a specific disease but a deformity complicating many diseases. Curve progression is the major concern irrespective of the initiating cause. Idiopathic scoliosis is arguably postural in nature and in some subjects develops from intrauterine compression. Analysis of the pathogenesis leads to the conclusion that progression is due to an accelerated premature osteoarthrosis induced by insidious tissue fatigue of biomechanical origin. The chronic cumulative effect of repetitive tensile stresses applied asymmetrically to the postural deformity, manifested by loss of tensile strength and tissue cohesion, leads to fragility and eventual tissue disintegration of vertebrae, intervertebral discs, and laxity of ligaments. Early treatment, prevention, and avoidance of stresses that accentuate progression are of paramount import. PMID- 12645633 TI - Monoclonality in Helicobacter pylori-positive gastric biopsies: an early detection of mucosa-associated lymphoid tissue lymphoma. AB - Mucosa-associated lymphoid tissue (MALT) is not present in healthy gastric mucosa, but it can develop in sites of long-persisting inflammation and is connected with the development of MALT lymphoma. A monoclonal lymphocyte population is one of the characteristics of such lymphomas. In this study we analyzed gastric biopsies (formalin fixed and paraffin embedded or frozen) in 93 patients with dyspepsia accompanied by Helicobacter pylori infection. We applied PCR and single-cell immunocytochemistry to detect the clonality of the gastric B cell population. Immunocytochemistry performed on 33 frozen biopsies showed two samples with monoclonal pattern. PCR analysis of immunoglobulin heavy-chain (IgH) gene rearrangements revealed two monoclonal populations out of 161 biopsies from 60 patients. We conclude that PCR analysis was the most sensitive method, which gave us insight into the nature of the earliest stage of MALT lymphoma in gastric biopsies. PMID- 12645634 TI - Inhibition of LPS-induced nitric oxide production in RAW cells by radioprotective thiols. AB - Nitric oxide (NO) is involved in producing damage after exposure to radiation and also in the toxicity associated with bacterial endotoxin (lipopolysaccharide, LPS). We have evaluated different radioprotective thiols for their effects on LPS stimulated NO production in mouse macrophage cells, RAW 264.7. Our results indicate that although thiols inhibited NO production in general, the degree of inhibition depended upon the thiol compound. Long-chain aminothiols like WR-1065 [N-(2-mercaptoethyl)-1,3-diaminopropane] exerted a strong inhibition; but its parent drug, amifostine, which protects mice against radiation lethality, was not as effective as WR-1065. Diethyl dithiocarbamate, which is less effective than amifostine as a radioprotector, strongly inhibited NO production from macrophages. These results indicate that the radioprotective potential of sulfhydryl compounds is not related to its ability to inhibit NO production by macrophages and suggest that some of the thiol radioprotectors may effectively ameliorate the fatal symptoms of hypotensive shock, associated with endotoxin (LPS)-induced NO production. PMID- 12645636 TI - Background and overview of a symposium to update the biology of atherosclerosis and its implications for treatment strategies. PMID- 12645635 TI - Hepatotoxin N-nitrosomorpholine-induced carcinogenesis in rat liver: ex vivo exploration of preneoplastic and neoplastic hepatocytes. AB - N-nitrosomorpholine (NNM) is a hepatotoxic and hepatocarcinogenic agent. This agent was administered in the form of drinking water which contained 200 mg of NNM/liter. Its time-dependent intake profile showed four phases over 20 weeks, followed by a fifth phase where only water was supplied. Most frequently, hepatocellular carcinoma appeared between the end of phase IV and the beginning of phase V. At 5 weeks of NNM administration, foci of altered hepatocytes (FAH) containing 100-1000 hepatocytes could be isolated together with free hepatocytes by the collagenase perfusion method. When these foci were grown on the William's Medium E containing hormonally defined medium, they were able to survive approximately twice as long as normal hepatocytes At 10 weeks of NNM administration, few FAH were isolated together with free hepatocytes. The hepatocytes which had been placed under extended chemical stress showed increased heat tolerance (7 to 8 h) at 43 degrees C, while normal hepatocytes could survive 3 to 4 h. At the neoplastic phase spanning the end of the 20 weeks of the NNM administration and water phase, the rats bearing hepatocellular carcinoma entered the terminal stage, where observable tumor masses could be isolated from the tumor bearing liver and tested for ex vivo growth in tissue culture. After stabilization of the isolated primary hepatoma cells through 10 passages of propagation on William's Medium E or minimal Eagle's medium containing 10% FBS, their gene expression profile was analyzed by DNA microarray and compared with the profile of normal hepatocytes. The comparison revealed that upregulation involved ribosome-dependent protein synthesis, including 40S ribosomal proteins (S4, S7, S18, S20), 60S ribosomal proteins (L6, L21, L32, L37, P1), initiation factor 4A, and elongation factor 1alpha. PMID- 12645637 TI - Vascular biology of atherosclerosis: overview and state of the art. AB - Our current understanding of the vascular biology of atherogenesis and its clinical manifestations suggests a pathophysiology that is much more complex than mere lipid storage. Recent advances support the current view of atherosclerosis as an inflammatory process that initiates and promotes lesion development to the point of acute thrombotic complications and clinical events. Inflammatory cells localize in early-stage atherosclerotic lesions, and recent basic research has established a causal relation between inflammatory mediators or cytokines, and the steps involved in progressing from local inflammation through plaque formation. Inhibition of the action of certain specific proinflammatory cytokines, such as CD40 ligand, interferes with atherogenesis in mice. Increased circulating levels of inflammatory markers indicate increased cardiovascular risk. Thus, the time has come to embrace inflammation as a common pathway for atherogenic risk factors and for providing new opportunities for therapeutic intervention. PMID- 12645638 TI - Role of oxidative stress in atherosclerosis. AB - The common risk factors for atherosclerosis increase production of reactive oxygen species (ROS) by endothelial, vascular smooth muscle, and adventitial cells. These ROS initiate processes involved in atherogenesis through several important enzyme systems, including xanthine oxidase, nicotinamide adenine dinucleotide phosphate (NADPH) oxidases, and nitric oxide synthase. Physical forces also regulate vascular production of ROS. Oscillatory shear, which is present at sites where atherosclerosis develops, seems a particularly potent stimulus of superoxide production. The signaling cascade for activation of the NAD(P)H oxidase by angiotensin II has recently been elucidated and seems to involve a feed-forward mechanism that permits ongoing production of ROS for prolonged periods. Oxidative stress in humans with coronary artery disease is also exacerbated by a reduction of vascular extracellular superoxide dismutase, normally an important protective enzyme against the superoxide anion. PMID- 12645639 TI - Oxidative stress: new approaches to diagnosis and prognosis in atherosclerosis. AB - Oxidative modifications of low-density lipoprotein (LDL) have been proposed to play a critical role in atherogenesis. To test the role of proposed antioxidants in inhibiting LDL oxidation and vascular disease, it is important to identify the biologically relevant sources of oxidative stress in the human arterial wall. Mass spectrometric (MS) quantification of oxidized amino acids in proteins was used as a "molecular fingerprint" to identify the pathways that inflict oxidative damage in vivo. For example, myeloperoxidase is expressed in macrophages in human atherosclerotic lesions, and immunohistochemical studies suggest that it might be a pathway for LDL oxidation. We found that hypochlorous acid, tyrosyl radical, and reactive nitrogen species generated by myeloperoxidase each yielded a unique pattern of protein oxidation products in vitro. MS analysis of human atherosclerotic tissue revealed a similar pattern of oxidation products. This strategy has pinpointed myeloperoxidase as a pathway that promotes LDL oxidation in the human artery wall. It is noteworthy that vitamin E fails to inhibit LDL oxidation by myeloperoxidase in vitro. Because the utility of an antioxidant depends critically on the nature of the oxidant that inflicts tissue damage, interventions that specifically inhibit physiologically relevant pathways would be logical candidates for clinical trials of antioxidants. Such a rational approach to therapy is likely to accelerate progress against oxidative stress and coronary artery disease. PMID- 12645640 TI - New understanding, diagnosis, and prognosis of atherothrombosis and the role of imaging. AB - Despite crucial advances in our knowledge of the pathologic mechanisms and the availability of effective diagnostic and treatment modalities, coronary atherothrombosis remains the most frequent cause of ischemic heart disease. Plaque disruption with superimposed thrombosis is the main cause of unstable angina, myocardial infarction, and sudden death. New findings have recently introduced exciting concepts that could have major impact on the treatment of the atherothrombotic disease. We will discuss the mechanisms that lead to the development of atherothrombosis and those responsible for the acute coronary syndromes, as well as some of the concepts derived from in vivo observations using new imaging technologies (eg, high-resolution magnetic resonance imaging). PMID- 12645641 TI - New frontiers and unresolved controversies in percutaneous coronary intervention. AB - Percutaneous coronary intervention (PCI) has become a key modality of therapy of patients with coronary artery disease. However, with the exception of acute coronary syndromes, there is as yet no conclusive evidence from randomized controlled trials that PCI improves survival and outcome compared with optimal medical therapy. Randomized trials comparing PCI and coronary artery bypass grafting in patients with stable coronary artery disease conducted before stenting showed superiority of the surgical approach. Early results from stent trials continue to indicate better symptom relief and less need for repeat intervention with the surgical approach, although myocardial infarction and mortality are similar with the 2 revascularization modalities. In the setting of acute coronary syndromes, however, recent trials suggest superiority of an aggressive approach, including PCI, compared with a conservative strategy of medical therapy only. This benefit extends to patients with acute ST-elevation myocardial infarction, including those with shock. In the latter setting, mortality remains high and novel approaches to protect myocardial metabolism and improve perfusion will be needed to further improve survival. The role of revascularization in patients with completed myocardial infarction and persistently occluded arteries is much less clear. Although the late open artery hypothesis is widely accepted and interventional cardiologists routinely open late infarct-related coronary occlusions, the available evidence to justify this strategy is weak. New devices, such as thrombectomy devices in thrombus-laden lesions and drug-eluting stents, offer promise, although all lack the degree of proof of benefit and safety typically required of new pharmaceutical agents. As new technologies and therapies with the potential to improve outcomes and decelerate the atherosclerotic process are developed, it will be important for investigators and industry to collaborate in incorporating these advances into ongoing and new trials testing the efficacy of PCI in all settings. PMID- 12645642 TI - Chemistry and pharmacology of vascular protectants: a novel approach to the treatment of atherosclerosis and coronary artery disease. AB - This review addresses the role of oxidative stress in the pathology of atherosclerosis and why it is now believed that atherosclerosis is not only a disease of oxidative stress but also of chronic inflammation. Perhaps more importantly, this review also describes the vascular protectant (V-protectant) technology platform originated at AtheroGenics, Inc., from which a series of inhibitory compounds has emerged to treat a number of chronic inflammatory diseases, including atherosclerosis. In atherosclerosis, these drugs not only act as antioxidants, but also as lipid modulators, inhibitors of inflammation, and inhibitors of gene expression. It is also important to understand the basis for considering vascular cell adhesion molecule-1 (VCAM-1) as a reduction-oxidation sensitive protein, which has a key role in the early phases of atherosclerosis. The review concludes with a description of the design and chemistry of AtheroGenics' lead clinical development compound, AGI-1067, and an analysis of its preclinical in vitro and in vivo profile. AGI-1067 is a novel, potent antioxidant with anti-inflammatory properties. It inhibits gene expression of VCAM-1 and monocyte chemoattractant protein-1, decreases low-density lipoprotein cholesterol levels, and prevents atherosclerosis in a number of animal models. AGI-1067 is currently undergoing clinical trials as an antiatherosclerotic agent. PMID- 12645643 TI - Clinical results with AGI-1067: a novel antioxidant vascular protectant. AB - A large body of evidence points to oxidative stress as an important trigger in the complex chain of events leading to atherosclerosis. Reactive oxygen species have also been implicated in the pathophysiology of restenosis after percutaneous coronary interventions (PCI). The powerful antioxidant probucol has been shown to prevent coronary restenosis after balloon angioplasty in the MultiVitamins and Probucol (MVP) trial and other clinical studies. Probucol has also induced regression of carotid atherosclerosis in the Fukuoka Atherosclerosis Trial (FAST). However, prolongation of the QT interval with probucol remains a long term safety concern. AGI-1067, a metabolically stable analog of probucol, is a vascular protectant (V-protectant) with strong antioxidant properties, equipotent to those of probucol. This V-protectant has been effective at preventing atherosclerosis in all tested animal models, including the low-density lipoprotein receptor-deficient and apolipoprotein E-knockout mice and the hypercholesterolemic primate. AGI-1067 improved luminal dimensions of the PCI site and reduced restenosis in the Canadian Antioxidant Restenosis Trial (CART 1). In contrast to probucol, AGI-1067 did not induce prolongation of the QT interval. AGI-1067 also improved luminal dimensions of the reference segments in the PCI vessels in CART-1, an effect that suggests a direct antiatherosclerosis effect. This has potentially important implications, as local approaches to prevent restenosis, such as coated stents, are not expected to prevent atherosclerosis progression, myocardial infarction, and cardiovascular death. Considering that oxidative stress and inflammation may persist for a prolonged period after stenting, treatment with AGI-1067 for the entire period of risk after PCI (instead of only 4 weeks in CART-1) may result in enhanced protection against luminal renarrowing in the ongoing multicenter CART-2 trial. Because the ultimate goal of therapy for patients with coronary artery disease must remain prevention of disease progression and atherosclerosis-related events, CART-2 will test the value of AGI-1067 for the reduction of both post-PCI restenosis and atherosclerosis progression. PMID- 12645644 TI - State of the art in coronary intervention. AB - Issues related to restenosis have been known since the beginning of coronary intervention, and by now, restenosis has been characterized in terms of its time course, pathophysiology, clinical presentation, and some of its histology. In 1984, the initial National Heart, Lung, and Blood Institute's percutaneous transluminal coronary angioplasty registry reported a restenosis rate of 33.6% and identified male gender, new unstable angina, diabetes mellitus, and treatment of bypass graft stenoses as risk factors. Today restenosis still occurs in 33% of patients, is still associated with recurrent angina, and occurs usually within several months after a successful intervention. These current conclusions are very interesting because we now treat a clearly different patient population. Clinical trials have documented that restenosis results in adverse clinical consequences. In recent large clinical trials involving patients with class B or C lesions, pharmacologic intervention with such agents as tranilast was still associated with a 33% restenosis rate and had no significant effect on major adverse cardiac events. More recently, drug-coated stents have been introduced and are undergoing testing in large clinical trials to definitively establish the long-term efficacy and safety suggested in their early promising experience. Thus, whether the solution for restenosis is at hand or whether we will continue to see patients who undergo successful intervention and develop restenosis only months later in the some position as the initial lesion remains an open issue. PMID- 12645645 TI - Fragile sites and bladder cancer. AB - Continued reports of associations between environmentally induced chromosomal fragile sites and cancer prompted us to undertake a review of current literature to examine whether there might be a relationship between fragile sites and chromosomal alterations reported for bladder cancer. It was found that more than half (56%; odds ratio [OR] = 4.70) of chromosomal rearrangements reported for bladder cancer were located at 77 (65%) of the 118 recognized fragile sites (OR = 6.88). Furthermore, 55% of the fragile sites implicated coincided with one or more genes that have been associated with human cancer (such as oncogenes, tumor suppressor, relonc, transloc, disorder, apoptotic, and angiogenic genes). The most common fragile sites involved were FRA1D, FRA1F, FRA8C, FRA9D, FRA9E, and FRA11C. This correlation suggests that there may be profiles of genetic damage via fragile site expression that lead to the development of at least a proportion of bladder cancers. PMID- 12645646 TI - A consistent region of deletion on 1p36 in meningiomas: identification and relation to malignant progression. AB - We analyzed the genetic aberrations on chromosome arms 1p, 10q, and 14q, which are thought to be loci that include putative tumor suppressor genes in meningiomas. We initially conducted molecular genetic testing on a total of 72 tumors including 15 atypical and 8 anaplastic meningiomas using double-target fluorescence in situ hybridization. An incidence of deletion of 1p was observed in 16.3% of histologically benign, 86.7% of atypical, and 87.5% of anaplastic meningiomas. Microsatellite analysis for loss of heterozygosity on 1p, 10q, and 14q was performed in 15 tumors (6 benign, 6 atypical, and 3 anaplastic meningiomas). We detected alimited deleted region on 1p36 in two tumors and suggest a new consistent region of deletion at 1p36.21 approximately p23 distal to D1S507 and proximal to D1S214, which spans 8.21 megabases. In addition, loss of 10q was detected in two of three secondary atypical meningiomas, and loss of 14q in two of three primary anaplastic meningiomas. We suggest that one of the putative suppressor genes is located at 1p36.21 approximately p23, and that 10q loss may contribute to the malignant progression from benign to atypical meningiomas. PMID- 12645647 TI - Molecular cytogenetic parameters in Ewing sarcoma. AB - To evaluate possible genomic instability and possible random aneuploidy, we applied comparative genomic hybridization and fluorescence in situ techniques, and evaluated telomerase activity in 16 cases of Ewing sarcoma (EWS) and compared the results to 7 controls. Common secondary aberrations (gains of chromosomes 8 and 12) were found in the study group. There was a direct correlation between the detection of random aneuploidy and development of tumor relapse (P = 0.0047). Other detectable abnormal parameters (secondary) and high telomerase activity were also more common among the cases with relapse but did not reach a statistical significance (probably because of the small sample size). In EWS, the detection of random aneuploidy seems to be a sensitive parameter in the prediction of tumor relapse. PMID- 12645648 TI - Promoter hypermethylation of DLC-1, a candidate tumor suppressor gene, in several common human cancers. AB - Aberrant methylation of CpG islands within the promoter regions of tumor suppressor or cancer-related genes is a common mechanism leading to the silencing of gene expression. To determine whether aberrant methylation is a contributing factor to transcriptional inactivation of DLC-1 (deleted in liver cancer-1), a candidate tumor suppressor gene, we examined its methylation status in twelve hepatocellular carcinoma. breast, colon, and prostate tumor cell lines with low or undetectable expression of DLC-1. By Southern blot analysis of DNA digested with the methylation sensitive enzyme HpaII, we found a different degree of promoter hypermethylation in all cell lines with aberrant DLC-1 expression. The hypermethylation status was reversed by the addition of 5-aza-2'-deoxycytidine, a demethylating agent, in one human hepatocellular carcinoma line. These observations suggest that hypermethylation is responsible for abrogating the function of the DLC-1 gene in a subset of liver, breast, colon, and prostate cancers. PMID- 12645649 TI - Karyotypic abnormalities in myelofibrosis following polycythemia vera. AB - Polycythemia vera (PV) is a chronic myeloproliferative disease characterized by an increase of total red cell volume; in 10% to 15% of cases, bone marrow fibrosis complicates the course of the disease after several years, resulting in a hematologic picture mimicking myelofibrosis with myelocytic metaplasia (MMM). This condition is known as post polycythemic myelofibrosis (PPMF). Among 30 patients with PPMF followed in Northern France, 27 (90%) expressed one or two abnormal clones in myelocytic cell cultures. Of these, 19 (70%) had partial or complete trisomy 1q. This common anomaly either resulted from unbalanced translocations with acrocentric chromosomes, that is, 13, 14, and 15, or other chromosomes, that is, 1, 6, 7, 9, 16, 19, and Y, or from partial or total duplication of long arm of chromosome 1. A single patient had an isochromosome 1q leading to tetrasomy 1q. In all cases, a common trisomic region spanning 1q21 to 1q32 has been identified. Given that most patients had previously received chemotherapy or radio-phosphorus to control the polycythemic phase of their disease, this study illustrates the increased frequency of cytogenetic abnormalities after such treatments: 90% versus 50% in de novo MMM. Moreover, karyotype can be used to distinguish PPMF-where trisomy 1q is the main anomaly from primary MMM where trisomy 1q is rare and deletions 13q or 20q are far more common. Whether trisomy 1q is or is not a secondary event remains a matter of debate, as well as the role of cytotoxic treatments. PMID- 12645650 TI - Molecular cytogenetic characterization of nasopharyngeal carcinoma cell lines and xenografts by comparative genomic hybridization and spectral karyotyping. AB - Nasopharyngeal carcinoma (NPC) cell lines and xenografts represent valuable models for functional and therapeutic studies on this common malignancy in Southeast Asia. The karyotypic information in most NPC cell lines and xenografts, however, remains largely unclear to date. We have characterized the chromosomal aberrations in six commonly used human NPC cell lines and xenografts using the molecular cytogenetic technique of comparative genomic hybridization (CGH). Genomic imbalances identified in cell lines were further correlated with structural abnormalities indicated from spectral karyotyping (SKY) analysis. CGH revealed consistent overrepresentations of 8q (six out of six cases) with a smallest overlapping region identified on 8q21.1 approximately q22. Other common gains included 7p (4/6 cases), 7q (4/6 cases), 12q (4/6), and 20q (4/6 cases), where minimal overlapping regions were suggested on 7p15 approximately p14, 7q11.2 approximately q21, and 12q22 approximately q24.1. Common losses were detected on 3p12 approximately p21 (4/6 cases) and 11q14 approximately qter (4/6 cases). Although SKY analysis on cell lines revealed predominantly unbalanced rearrangements, reciprocal translocations that involved chromosome 2 [i.e., t(1;2), t(2;3), and t(2;4)] were suggested. Furthermore, SKY examination illustrated additional breakpoints on a number of apparently balanced chromosomes. These breakpoints included 3p21, 3q26, 5q31, 6p21.1 approximately p25, 7p14 approximately p22, and 8q22. Our finding of regional gains and losses and breakpoints represents information that may contribute to NPC studies in vitro. PMID- 12645651 TI - Are gains of chromosomal regions 7q and 11p important abnormalities in neuroblastoma? AB - Neuroblastoma exhibiting deletion of a segment of the long arm of chromosome 11 represents a genetic subtype of tumor that is distinct from those exhibiting MYCN amplification or 1p deletion. The 11q- genetic subtype is further characterized by gain of 17q and loss of distal 3p material. Gain of 11p material has also been reported in neuroblastoma with 11q loss, but at a considerably lower frequency than gain of 17q or loss of the distal 3p region. Our results, however, indicate that gain of 11p may occur more frequently in 11q- neuroblastoma than what was previously realized. Comparative genomic hybridization analyses of neuroblastoma tissue from eleven patients indicated that six of 11 tumors (55%) with loss of 11q also possessed gain of 11p. The shortest region of 11p gain was 11p11.2- >p14. G-banding and fluorescence in situ hybridization analysis performed on tumor cells from primary and metastatic sites from one patient allowed us to infer that gain of 11p arose secondarily to the abnormality that led to the loss of 11q material. Gain of an entire chromosome 7 was detected in 17 of 43 (40%) tumors, whereas gain of 7q was detected in 5 of 43 (12%) tumors. Unlike gain of 11p, gain of an entire chromosome 7 appears to be prevalent in all tumor stages and is not limited to the 11q- tumor subtype. Gain of 7q, however, is more prevalent in higher stage tumors. G-band cytogenetic analysis indicated that an unbalanced t(3;7) was responsible for the gain of 7q and loss of 3p material in one case. We discuss the possibility that gain of 7/7q, and 11p material may contribute to either tumorigenesis or progression. PMID- 12645652 TI - A unique clone involving multiple structural chromosome rearrangements in a myelodysplastic syndrome case. AB - In a young female patient presenting with a myelodysplastic syndrome (MDS), a unique clone involving six structural chromosome rearrangements was identified using G-banding and molecular cytogenetic techniques. Fifty GTG-banded metaphases from bone marrow were initially analyzed and all metaphases contained all of the six structural chromosome rearrangements. To further define the GTG-banded karyotype, a series of fluorescence in situ hybridization and primed in situ labeling experiments were performed and the karyotype was then characterized as: 46,XX,r(5)(p13q13),der(20)t(5;20),dup(11)(p11.2p15), r(11)(p15q25),del(13)(q14),idic(22)(p11). The patient quickly progressed to acute nonlymphocytic leukemia three months after the diagnosis and died of a hemorrhage in the brain parenchyma two months later. In this case, the multiple structural chromosome rearrangements conferred an obvious cellular proliferative advantage and indicated a very poor prognosis. Considering that multiple chromosome abnormalities associated with MDS transformation are often polyclonal, this unique clone involving six structural chromosome rearrangements make our case highly unusual. PMID- 12645653 TI - Chromosomal alterations in osteosarcoma cell lines revealed by comparative genomic hybridization and multicolor karyotyping. AB - We characterized the chromosomal alterations in eight osteosarcoma cell lines (OST, HOS, U-2 OS, ZK-58, MG-63, SJSA-1, Saos-2, and MNNG) by comparative genomic hybridization (CGH); gains and losses of DNA sequences were defined as chromosomal regions with a fluorescence ratio, wherein all of the 95% confidence interval was above 1.25 and below 0.75, respectively. In four of 8 cell lines, multicolor karyotyping (MK) was added. CGH revealed the average number of aberrations per cell line was 20.8 (range: 10-31); the average numbers of gains and losses were 11.1 and 9.6, respectively. The frequent gains were identified on 1p21 approximately q24, 1q25-q31, 7p21, 7q31, 8q23 approximately q24, and 14q21; frequent losses were at 18q21 approximately q22, 18q12, 19p, and 3p12 approximately p14. High-level gains were observed on 8q23 approximately q24, 5p, and 1p21 approximately p22. MK revealed the most common translocations in the four cell lines were t(8;9), t(1;3), t(3;5), t(1;13), t(2;6), t(3; 17), t(1;15), t(10;20), and t(6;20). Chromosomes 1, 3, 8, 9, and 20 were most frequently involved in translocation events. The concordance rate of aberrations in CGH and translocations in MK was 76%. MK was useful to identify the chromosomal alterations and as a supplement to the CGH results in three of four chromosomes. PMID- 12645654 TI - Generation of the NUP98-TOP1 fusion transcript by the t(11;20) (p15;q11) in a case of acute monocytic leukemia. AB - A rare chromosomal translocation, (11;20)(p15;q11), was detected in a 29-year-old male patient diagnosed with acute monocytic leukemia (AMoL) according to the French-American-British classification criteria. Whole chromosome painting analysis with paints for chromosomes 11 and 20 confirmed the result of conventional cytogenetic analysis. Reverse transcriptase polymerase chain reaction revealed the NUP98-TOP1 fusion transcript. To our knowledge, this is the second report of the translocation involving NUP98 and TOP1 genes in AMoL. On reviewing the literature, we suggest that t(11;20)(p15q11) is associated with myelocytic disorders rather than lymphocytic proliferative diseases. PMID- 12645655 TI - MYCN gain and MYCN amplification in a stage 4S neuroblastoma. AB - Stage 4S neuroblastoma is a disease associated with spontaneous regression and good survival. We present a patient whose evolution has shown the variety and complexity of this disease in infants. Biologic factors, such as ploidy, MYCN copy number, loss of 1p36, and other chromosomal gains and losses were determined. A complex pattern of genetic abnormalities, such as near-diploidy, MYCN gain (2-4 copies per haploid genome) and imbalance/deletion of 1p36 was seen in the diagnostic sample. An extensive disseminated disease after a latent period of 26 months was associated with a special genetic evolution, such a tetraploidy, MYCN amplification (2:100-500 copies), 1p36 deletion, and gain of 17q. Our results provide evidence that either the primary tumor was heterogeneous in terms of gene amplification or that amplification was acquired later on as a transition from MYCN gain. We suggest that near-di-/tetraploid 4S tumors with MYCN gain and/or deletion 1p could be progressing 4S tumors. PMID- 12645656 TI - Infrequent but high-level amplification of 17p11.2 approximately p12 in human glioma. AB - We reported previously the amplification of DNA markers in 17p12 in 3 of 60 high grade gliomas. To detect additional cases, we screened in total 104 gliomas of various types and grades by Southern blot analysis using marker 745R, which is within the commonly amplified region. However, no other caseswith significant amplification (amplification level > 4) were found. To investigate in detail the extent of the amplifications in the three tumors, which were all glioblastomas, we determined 17p11.2 approximately p12 amplification profiles by semiquantitative polymerase chain reaction using 15 microsatellite markers and seven candidate genes. Distinct and high-level amplifications, with maximum levels ranging from 15 to 38, were found in these tumors. The 0.8 Mb-region between D17S1525 and MAP2K4 in 17p12 proved to be commonly amplified in these tumors. In one tumor, a heterogeneous distribution of the amplification in 17p12 was found, suggesting that it is a late event during glioma tumorigenesis. Another tumor showed additional high-level amplification of PMP22 and D17S1843 in 17p11.2. From the high-level amplifications we conclude that at least one, but possibly more, putative oncogenes are present in 17p11.2 approximately p12 whose amplifications and/or overexpressions contribute to glioma tumorigenesis. PMID- 12645657 TI - Successful treatment with cyclosporin A of myelodysplastic syndrome with erythroid hypoplasia associated with t(6;8)(q15;q22). AB - We report a t(6;8)(q15;q22) in a patient with myelodysplastic syndrome (MDS) with erythroid hypoplasia. The patient was successfully treated with an immunosuppressive treatment with cyclosporin A, while the translocation was repeatedly detected as the sole anomaly with the percentages of positive cells ranging from 5% to 70%. To our knowledge, the t(6:8) has never been described in MDS. PMID- 12645659 TI - Acute B-cell lymphoblastic leukemia with t(14;18): establishment of a new cell line. PMID- 12645658 TI - Trisomy 21 and other chromosomal abnormalities in acute promyelocytic leukemia. AB - We describe a case of acute promyelocytic leukemia (APL) with t(15;17)(q22;q12) and trisomy 21 as an additional change in a patient who died at relapse after achieving complete remission (CR) for the duration of 20 months. A survey of 42 cases of APL with cytogenetic study performed at our institutionover the past 10 years showed 12 cases (28.6%) having chromosomal changes in addition to t(15;17). Trisomy 8 and trisomy 21 as additional changes were noted in 4 and 2 cases, respectively, with one patient showing both trisomies simultaneously. Two cases showed t(15;17) in hyperdiploid clones. Among the 10 patients with follow-up data, all eventually relapsed and none achieved continuous complete remission 1. Survival analysis performed in APL patients with adequate follow-up data showed no significant difference in overall and disease free survival between those with and without additional cytogenetic changes. After excluding cases with one induction death, the overall survival was significantly in favor of the group without additional cytogenetic abnormalities (P = 0.022). Late relapses may therefore be significantly more common in APL patients with additional cytogenetic abnormalities, and may not be reflected by analysis focused at three year survival only. As APL is now considered a curable disease, any confirmed long-term survival impact of additional cytogenetic changes is expected to have important management implications. PMID- 12645660 TI - Minimal residual disease and trisomy 8. PMID- 12645662 TI - Defining the glycophenotype of squamous epithelia using plant and mammalian lectins. Differentiation-dependent expression of alpha2,6- and alpha2,3-linked N acetylneuraminic acid in squamous epithelia and carcinomas, and its differential effect on binding of the endogenous lectins galectins-1 and -3. AB - A thorough characterization of the properties of squamous epithelial cells is necessary in order to improve our understanding of the functional aspects of normal development and malignant aberrations. Up to now, studies have focused almost exclusively on monitoring distinct protein markers. With our growing awareness of the coding function of glycan chains of cellular glycoconjugates and their interaction with receptors (lectins) in situ, defining the glycophenotype of these cells has become an important issue. Whereas the commonly applied plant lectins are tools used to map the presence and localization of biochemically defined saccharide epitopes, the introduction of endogenous (mammalian) lectins to this analysis enables us to take the step from monitoring the presence of glycan to understanding the functional implications by revealing ligand properties of the detected epitope for tissue lectin. Thus, in this study we investigated a distinct aspect of glycosylation using plant and mammalian lectins, i.e. the linkage type of sialylation. We first mapped the expression profile of the type of sialylation (alpha2,3- or alpha2,6-linked) by plant lectins. Based on the hypothesis that this factor regulates accessibility of ligands for endogenous lectins we introduced two labeled galectins to this study. Galectin-3 (but not galectin-1) binding was related to cell differentiation in normal adult and developing epithelia, cultured epidermal cells, and carcinomas derived from these epithelia. The presented data suggest that alpha2,6-linked N acetyl-D-neuraminic acid moieties could serve to mask galectin-3-reactive glycoepitopes. As a consequence, monitoring of the linkage type of sialic acid in glycans by plant lectins therefore has implications for the extent of glycan reactivity with endogenous lectins, pointing to a potential function of changes in sialylation type beyond these cell and lectin systems. PMID- 12645661 TI - Suppressors of cytokine signalling: SOCS. AB - The mechanism of positive regulation of cytokine signalling pathways has been well investigated, whereas our knowledge of negative regulation is relatively sparse. Here we review recent literature on important negative regulators: the family of suppressors of cytokine signalling, SOCS, consisting of eight members (SOCS-1 to SOCS-7 and CIS) all sharing a central SH2 domain and a C-terminal SOCS box. Expression of CIS, SOCS-1, SOCS-2 and SOCS-3 is induced by various cytokines, and overexpression studies in various cell lines have demonstrated their inhibitory roles. These family members have been implicated in the negative regulation of several pathways, particularly the JAK/STAT pathway, and since this signalling pathway is responsible for their induction, they form part of a classical negative feedback circuit. To date, at least three different modulating mechanisms have been demonstrated: through the SH2 domain they bind to phosphotyrosines on the target protein, leading to inhibition of signal transduction by N-terminal inactivation of JAK, by blocking access of STAT to the receptor sites, or by SOCS box-targeting bound proteins to proteasomal degradation. In gene modification studies in mice, it has been demonstrated that SOCS-1 plays an important role in IFNgamma-regulation and T-cell differentiation, while SOCS-2 seems necessary for normal growth regulation. SOCS-3(-/-) mice die during embryogenesis for a reason still not fully understood, but insufficient control of fetal erythropoiesis or defects in placental development may be involved. The physiological role for the other family members, as well as their molecular regulation mechanisms, remain to be revealed. PMID- 12645663 TI - Lack of lipoprotein-dependent effects on the cytotoxic interactions of Actinobacillus actinomycetemcomitans leukotoxin with human neutrophils. AB - A high odds ratio has been reported for hyperlipidemia and periodontal diseases in humans, and the severity of periodontitis seems to correlate with the hyperlipidemic status of the patients. Early studies indicated that the lipoprotein-containing fraction of the serum enhances the leukotoxic activity of the periodontopathogen Actinobacillus actinomycetemcomitans against human polymorphonuclear leukocytes (PMNL). The protease inhibitors of normal serum account for this enhancement, while delipidated serum has no effect on the leukotoxin-dependent PMNL cytolysis. No information exists for the effect of serum lipoproteins or hyperlipidemic serum. The aim of this study was to evaluate the role of serum lipoproteins in the interaction of the leukotoxin of A. actinomycetemcomitans with human PMNL. Purified leukotoxin was mixed with human PMNL prepared from venous blood of healthy subjects and various varying amounts of hyperlipidemic or delipidated serum, or purified serum lipoproteins. The cytolytic activity of leukotoxin was determined by activity of the cytosol enzyme lactate dehydrogenase released from injured PMNL. The degranulating activity of the toxin was measured through the release of the granule components elastase and lactoferrin. Normal human serum without leukotoxin-neutralizing antibodies caused a 4-fold enhancement of the leukotoxic activity when present at concentrations of 5-10% in the reaction mixture. Serum lipoproteins had no effect when added at concentrations that occur normally in serum. At high concentrations, purified low density and very low-density lipoproteins increased the leukotoxicity of the mixture. Nevertheless, hyperlipidemic serum prepared from a normal serum by the addition of autologous lipoproteins had no influence on the leukotoxin-caused cytolysis compared to the normal serum. Pre-incubation of PMNL for 1 h in hyperlipidemic or delipidated serum had no effect on the leukotoxin-induced degranulation of PMNL. The results indicate that the cytotoxic interactions of A. actinomycetemcomitans leukotoxin against human PMNL are not influenced by the presence of serum lipoproteins. PMID- 12645664 TI - Nosocomial Legionella pneumophila serogroup 5 outbreak associated with persistent colonization of a hospital water system. AB - An outbreak of infections caused by Legionella pneumophila serogroup 5 was detected in a university hospital, and nosocomial reservoirs of the legionella epidemic were examined. Clinical isolates from two patients who had been affected by the L. pneumophila serogroup 5 outbreak, and from another patient with a legionella infection caused by the same serogroup 3 years later, were compared to L. pneumophila serogroup 5 isolates from the hospital water supply by two molecular methods, amplified fragment length polymorphism (AFLP) analysis and random amplified polymorphic DNA analysis (RAPD). Genotyping confirmed the epidemiological linkage of the first two patients, and linked their infections with the hospital water supply. The third clinical strain, which was also linked to the hospital water, was very similar to the epidemic strain. Even though the water distribution system was sanitized (superheat and flush sanitation), the epidemic strain was shown to be persisting in the hospital water outlets several years after its initial discovery. PMID- 12645665 TI - Modified pulsed-field gel electrophoresis protocol for typing of enterococci. AB - Controlling the spread of vancomycin-resistant enterococci (VRE) is an important task in hospital epidemiology. Pulsed-field gel electrophoresis (PFGE) has become the golden standard for molecular epidemiological characterisation of enterococcal isolates. For separation of DNA fragments by PFGE, different electrophoresis conditions have been recommended, but none of these protocols allows a satisfactory separation of both small and large DNA fragments of enterococci simultaneously. In this study we have speeded up the preparation of chromosomal DNA and defined new electrophoresis conditions that enhance separation of small and large DNA fragments for subtyping of enterococci with a 24 h PFGE. PMID- 12645666 TI - Primary marginal zone B-cell lymphoma of the lymph node resembling plasmacytoma arising from a plasma cell variant of Castleman's disease. A clinicopathological and immunohistochemical study of seven patients. AB - Nodal marginal zone B-cell lymphomas (NMZBL) occasionally represent prominent plasma cell differentiation. Recently, we presented a patient with NMZBL who exhibited histological features that resembled plasmacytoma arising from a localized plasma cell variant of Castleman's disease. To further clarify the clinicopathological, immunohistochemical, and genotypical findings, we studied seven such patients. Clinically, these patients were characterized by localized disease and an indolent clinical course with a slowly growing bulky mass in the affected lymph node. Only one patient exhibited paraproteinemia. Histologically, the lesions were characterized by numerous evenly distributed germinal centers in extensive sheets of plasma cells. Various numbers of centrocyte-like (CCL) cells arranged in a marginal zone distribution pattern occupied the peripheral region of the lymph node. The majority of the lymphoid follicles had atrophic or regressive germinal centers. A few lymphoid follicles were colonized by CCL cells. Immunohistochemistry showed that all of the lesions contained a monoclonal plasma cell population. In three tumors, a number of the CCL cells had a similar light chain restriction pattern to that observed in plasma cells. Two of the four patients evaluated exhibited clonal bands for the IgH gene by polymerase chain reaction assay. Moreover, the presence of surface IgM+, IgD- and CD27+ CCL- cells suggests that these tumors are derived from memory B-lymphocytes. PMID- 12645667 TI - Antibodies against beta-lactamase can improve ceftazidime treatment of lung infection with beta-lactam-resistant Pseudomonas aeruginosa in a rat model of chronic lung infection. AB - To test the hypothesis that antibodies against the chromosomal beta-lactamase of Pseudomonas aeruginosa (a beta ab) might act as beta-lactamase inhibitors in patients with cystic fibrosis and chronic lung infection with P. aeruginosa, we compared in a rat model of chronic lung infection the efficacy of treatment with ceftazidime in beta-lactamase-immunized (group I) and non-immunized (group II) rats. Chronic lung infection was established with alginate-embedded P. aeruginosa producing high amounts of beta-lactamase in 133 Lewis rats. Prior to infection, group I (66 rats) was immunized three times at 2-week intervals with purified beta-lactamase in incomplete Freund's adjuvant (IFA) and group II (67 rats) received IFA. Ceftazidime treatment was initiated after challenge and continued for 10 days, after which the rats were sacrificed and the lung bacteriology and pathology were analysed. Rat serum was analysed for the beta-lactamase inhibitory activity and a beta ab-specific IgG and IgG subclasses titres. Beta-lactamase inhibitory activity was found only in sera of rats belonging to group I and it was used to divide these rats into two subgroups: rats whose sera inhibited > or = 75% of beta-lactamase activity (responders) and rats whose sera inhibited < or = 25% of beta-lactamase activity (non-responders). The responder subgroup had significantly smaller pathological areas in the lungs and lower cfu/ml lung homogenate compared to the non-immunized group (p=0.02 and p=0.01, respectively) and compared to the non-responder subgroup (p=0.008 and p=0.0001, respectively). On the day of challenge, significantly higher titres of a beta ab-specific IgG and IgG subclasses antibodies were found in the responders compared to the non responders (p<0.0001). In the responder subgroup the avidity of IgG a beta ab was significantly higher than in the non-responder subgroup (p=0.0003). Our study showed that a beta ab with beta-lactamase inhibitory activity raised by immunization with beta-lactamase can improve the outcome of treatment with ceftazidime of resistant P. aeruginosa in a rat model of chronic lung infection. PMID- 12645668 TI - Oral fibroblasts produce more HGF and KGF than skin fibroblasts in response to co culture with keratinocytes. AB - The production of hepatocyte growth factor (HGF) and keratinocyte growth factor (KGF) in subepithelial fibroblasts from buccal mucosa, periodontal ligament, and skin was determined after co-culture with keratinocytes. The purpose was to detect differences between the fibroblast subpopulations that could explain regional variation in epithelial growth and wound healing. Normal human fibroblasts were cultured on polystyrene or maintained in collagen matrix and stimulated with keratinocytes cultured on membranes. The amount of HGF and KGF protein in the culture medium was determined every 24 h for 5 days by ELISA. When cultured on polystyrene, the constitutive level of KGF and HGF in periodontal fibroblasts was higher than the level in buccal and skin fibroblasts. In the presence of keratinocytes, all three types of fibroblasts in general increased their HGF and KGF production 2-3 times. When cells were maintained in collagen, the level of HGF and KGF was decreased mainly in skin cultures. However, in oral fibroblasts, induction after stimulation was at a similar level in collagen compared to on polystyrene. Skin fibroblasts maintained in collagen produced almost no HGF whether with or without stimulation. The results demonstrate that the secretion of KGF and HGF in both unstimulated fibroblasts and in fibroblasts co-cultured with keratinocytes is dependent on the type of fibroblasts. In general, the periodontal fibroblasts had the highest level of cytokine production. This high level of growth factor production may influence the proliferation and the migration of junctional epithelium and thereby influence the development of periodontal disease. PMID- 12645669 TI - Early dissemination rates of Friend murine leukaemia virus variants correlate with late pathogenesis. AB - FIS-2, a less oncogenic, immunosuppressive variant of the Friend murine leukaemia virus (F-MuLV), was used to explore whether the differences in biological features were related to early virus dissemination rates or sites of replication. We found that erythroblasts were the primary target cells for both F-MuLV and FIS 2, while B- and T-cells were infected later in the infection. Although FIS-2 replicated to similar titres as F-MuLV, we observed a delay in peak viraemia titre and in the number of virus-positive cells in bone marrow and spleen. Studies including the chimeric viruses RE3 (FIS-2LTR with a F-MuLV background) and RE4 (F-MuLV LTR with a FIS-2 background) indicated that the delay in dissemination was due to mutations in FIS-2 LTR. The kinetics for early virus replication correlated with previously reported mean latency time for virus induced erythroleukaemia in mice inoculated as newborns and with the onset of immunosuppression in adult mice. In addition, F-MuLV-induced late immunosuppression coincided with signs of erythroleukaemia and persistent viraemia. FIS-2 induced a more moderate late immunosuppression without persistent viraemia or signs of erythroleukaemia. Overall, our findings indicated that early viral replication is a prognostic factor in murine retrovirus-induced pathogenesis. PMID- 12645670 TI - Neurocognitive correlates of schizotypy in first degree relatives of schizophrenia patients. AB - We examined neurocognitive correlates of three dimensions of schizotypy in 63 healthy first degree relatives of schizophrenia patients. Neurocognitive measures of attention, verbal memory, and prefrontal functioning were combined with self report and interview measures of schizotypy. State-psychopathology (anxiety and depression) was a strong predictor for positive schizotypy (PS) and negative schizotypy (NS). PS was slightly correlated to verbal long-term memory, therefore weakly supporting the hypothesis that temporal-limbic malfunctioning underlies PS. NS was not correlated to any prefrontal measure, and therefore no evidence was found for the hypothesis that prefrontal malfunctioning underlies NS. Disorganization schizotypy (DS) was strongly correlated to the false alarm variable of the Continuous Performance Test (CPT), probably supporting the hypothesis of orbitofrontal malfunctioning underlying DS. This correlational pattern of DS echoes closely two schizophrenia studies reporting a relationship between formal thought disorder and the false alarm CPT variable. This similarity, across schizophrenia and relatives samples, may be considered as evidence that false alarms on the CPT and (subtle) problems in goal directedness of thinking are indicators of a genetically determined vulnerability to schizophrenia. PMID- 12645671 TI - Morbidity risk of psychiatric disorders among the first degree relatives of schizophrenia patients in Taiwan. AB - This study aimed to assess the boundaries of the schizophrenia spectrum and whether inclusion of such phenotypes increases power for linkage analysis of schizophrenia. Participants were 234 first degree relatives (FDRs) of 94 schizophrenia probands in Northern Taiwan who completed a direct interview using the Diagnostic Interview for Genetic Studies (DIGS). Based on best estimate diagnosis, the morbidity risk in the relatives for schizophrenia was 2.5 percent (Weinberg's shorter method) or 3.9 percent (Kaplan-Meier estimate). Depending on the stringency of diagnosis, lifetime prevalence was 2.6 percent to 4.7 percent for schizotypal personality disorder, 3.4 percent to 8.6 percent for paranoid personality disorder, and 1.3 percent to 3.4 percent for schizoid personality disorder. These figures are significantly higher than the corresponding figures in the general population. However, none of the recurrence risk ratio for any spectrum that included both schizophrenia and a personality disorder (3.0 to 5.9) was greater than that of schizophrenia alone (9.3 to 14.4). Thus, including schizophrenia-related personality disorders in the spectrum did not increase power for linkage analysis of schizophrenia. PMID- 12645672 TI - Clinical variables and genetic loading for schizophrenia: analysis of published Danish adoption study data. AB - Schizophrenia shows considerable clinical variation, but the relationship between clinical variables and degree of genetic loading for schizophrenia is unclear. We investigated this by analyzing published data from the adoption study of Kety et al. (1994) in Denmark. We sought to determine which clinical variables in proband adoptees with chronic schizophrenia predicted risk of schizophrenia in their biological relatives, using logistic regression analysis. We found that risk of chronic schizophrenia in relatives was predicted by the presence of pervasive negative symptoms (odds ratio [OR] = 9.44, 95% confidence interval [CI] = 1.98 45.01) and absence of pervasive positive symptoms (OR = 0.09, 95% CI = 0.01-0.78) in probands. Pervasive negative symptoms were defined by the presence of all of the symptoms: social withdrawal, autistic behavior, poverty of thought/speech, and flat affect. Pervasive positive symptoms were defined by the presence of all of the symptoms: suspiciousness/ideas of reference, delusions, auditory hallucinations, and other hallucinations. These clinical variables may be useful for refining phenotypic definitions of schizophrenia in molecular genetic studies. PMID- 12645673 TI - School performance as a premorbid marker for schizophrenia: a twin study. AB - Evidence is accumulating that developmental abnormalities, poor neuromotor function, and increased problem behavior precede the manifestation of schizophrenia itself. Information on school performance and behavioral development was obtained for 49 pairs of twins in which at least one twin suffered from schizophrenia (20 monozygotic [MZ], 29 dizygotic [DZ]) and for 43 pairs of healthy control twins (25 MZ, 18 DZ). Cox regression was used to analyze the contribution of schizophrenia, zygosity, sex, and year of birth to the age at which developmental divergence occurred. In both MZ and DZ twin pairs with schizophrenia, divergence in school performance occurred about 7.5 years earlier than it did in control twins, at 12 years of age, preceding the onset of psychosis by 10 years. This suggests that the first prodromal signs of schizophrenia manifest themselves as cognitive symptoms at the onset of puberty. Underperformance at school might therefore be considered one of the first signs of an until then latent vulnerability for schizophrenia. PMID- 12645674 TI - Symptomatology of the initial prodromal phase in schizophrenia. AB - The initial prodromal symptoms in schizophrenia were studied in 100 DSM-diagnosed patients and 100 controls. The median number of symptoms in the patients and the controls was 8 (range 2-13) and 0 (range 0-5), respectively. Patients developed symptoms indicating social, occupational, and affective dysfunction, whereas the controls' symptoms included magical content and disturbance in mood. There were significant differences in the frequency of several symptoms appearing in the subtypes. Initial prodromal symptoms were classified into negative, positive prepsychotic, and positive-disorganization categories. Patients with the disorganized subtype were more likely to have had negative symptoms in the prodromal state, and patients with the paranoid subtype were more likely to have had positive symptoms in the prodromal state. Observation of the course of symptoms from the prodromal to the psychotic state revealed that 58 percent of the symptoms showed increased intensity, 21 percent remained unchanged, 5 percent decreased, 3 percent evolved into other affective difficulties, 9 percent progressed into delusions, 1 percent progressed into hallucinations, and 3 percent disappeared. The Global Assessment of Functioning Scale showed that functioning is differentially affected among the subtypes even in the prodromal phase. These findings provide a better understanding of the initial prodromal state of schizophrenia, the signs and symptoms that best define it, and their prognostic significance. PMID- 12645675 TI - Attitudes of patients toward the first psychotic episode and the start of treatment. AB - Consecutively admitted patients with recent-onset schizophrenia disorders (n = 56) were interviewed about their awareness of mental disorder at first occurrence of psychotic symptoms, their views about the necessity of psychiatric treatment, their perception of the role of others in initiating psychiatric treatment, and suggestions they might have about getting treatment started at an earlier point in time. About 57 percent of the patients had at least some awareness of having a mental disorder at onset of psychotic symptoms, and 61 percent of the sample believed psychiatric help was unnecessary prior to the start of psychiatric treatment. A majority of the patients (91%) perceived others to be essential in initiating treatment. Most patients (82%) thought that no change is needed on the part of professional caregivers in order to facilitate early treatment. Some awareness of mental disorder at onset was related to a shorter duration of untreated psychosis (DUP). Delaying treatment until patients themselves become aware of the need for treatment may enlarge DUP. PMID- 12645676 TI - A comparison between schizophrenia patients and healthy controls on the expression of attentional blink in a rapid serial visual presentation (RSVP) paradigm. AB - The expression of attentional blink (AB) in 24 schizophrenia inpatients was compared to 22 healthy subjects in a dual-target rapid serial visual presentation (RSVP) paradigm in which a sequence of discrete stimuli was presented in rapid succession. Correct identification of the first target led to poorer detection of the second one when they were interspersed by distractors. This second-target deficit constitutes the AB effect, which is most pronounced between 200 and 500 ms after the offset of the first target stimulus and steadily decays as the number of intervening distractors increases. Despite relatively poor performance in terms of target identification within RSVP streams, schizophrenia patients expressed an AB effect that was as clear as that seen in healthy subjects. Moreover, there was evidence for an enhanced AB effect in schizophrenia patients. This outcome contrasts with the robust finding that schizophrenia patients are attenuated in the expression of prepulse inhibition, another paradigm believed to assess attentional control. The present results add to the extensive literature on the nature and specification of attentional dysfunction implicated in schizophrenia. PMID- 12645677 TI - Electrodermically nonresponsive schizophrenia patients make more errors in the Stroop Color Word Test, indicating selective attention deficit. AB - This study investigated whether skin conductance responsivity is associated with selective attention assessed by the Stroop Color Word Test (SCWT) in schizophrenia patients. The subjects (31 schizophrenia patients and 20 patients with other psychotic diagnoses) were selected from among inpatients of a general hospital psychiatric ward or day hospital attendees. They were matched with 31 healthy volunteers. The patients began experimental sessions immediately after remission of an acute episode. The three groups of participants were subdivided according to electrodermal responsivity into nonresponsive (NR) and responsive (R) groups. After the psychophysiological recording, the SCWT was applied. Results indicated that on the SCWT, the error interference of the NR schizophrenia group was significantly higher than that of all the other groups. Furthermore, the NR schizophrenia patients had significantly more negative symptoms than the R schizophrenia patients. These results suggest that there is a homogeneous subgroup of schizophrenia patients characterized by low neurovegetative responsiveness to external stimuli, predominance of negative symptoms, and selective attention deficit. PMID- 12645678 TI - Impaired temporal lobe processing of preattentive auditory discrimination in schizophrenia. AB - Feature-specific stimulus discrimination related to short-term auditory sensory memory can be studied electrophysiologically using a specific event-related potential (ERP) component termed mismatch negativity (MMN), which is generated in the auditory cortex, indexing automatic comparison of the existing memory trace to incoming novel stimuli. Previous results with electroencephalography (EEG) and magnetoencephalography (MEG) suggest that schizophrenia patients have attenuated MMN response and that preattentive auditory processing preceding MMN appears to be functionally asymmetric in schizophrenia. Here we studied parallel MMN activity of the hemispheres using a whole-head MEG by presenting stimulus blocks consisting of frequent standard and infrequent deviant tones to 15 schizophrenia patients and 19 healthy control subjects. Auditory evoked fields (AEFs) were recorded simultaneously over both auditory cortices. The equivalent current dipole (ECD) modeling revealed that patients had significant MMNm reduction (magnetic counterpart of MMN) in both temporal lobes. In addition, patients had significantly delayed MMNm in the left but not in the right hemisphere to ipsilateral auditory stimuli. These results suggest that patients with schizophrenia have impaired auditory processing in the temporal lobes underlying preattentive stimulus discrimination that is also selectively delayed in the left hemisphere. PMID- 12645679 TI - The language system in schizophrenia: effects of capacity and linguistic structure. AB - Dysfunction in receptive language processes has been reliably observed in individuals diagnosed with schizophrenia and their first degree family members. The present study addressed the unresolved issue of whether receptive syntax is intact in schizophrenia. The principal question concerned whether comprehension dysfunction in schizophrenia involves a disturbance in the processing of syntactic structure, a susceptibility to demands placed on temporal auditory processing, or some combination of these two general factors. Comprehension accuracy was compared between 32 males diagnosed with schizophrenia and 22 males with no lifetime diagnosis of psychiatric disorder. Accuracy was examined for responses to Who questions ("Who did X?" and "Who was done X?") about information in the sentential clauses (main vs. relative) of two types of relative sentences (subject-relatives vs. object-relatives) that were presented aurally at conversational and accelerated rates. The relationship between cognitive functions and comprehension accuracy was also tested. Results showed highly significant effects of diagnosis, syntactic structure, and temporal demand. Patients were characterized by reduced overall comprehension accuracy compared to controls. More important, patients and controls differed in their patterns of accuracy across the different types of syntactic structure. Finally, cognitive functions predicted but did not completely account for comprehension accuracy. Findings suggest the hypothesis that receptive syntax is disrupted in schizophrenia, and this dysfunction may not be entirely explained by compromised general cognitive ability. PMID- 12645680 TI - Differences in age of first hospitalization for schizophrenia among immigrants and nonimmigrants in a national case registry. AB - While previous studies have found an increased incidence of schizophrenia in some immigrant groups, differences in age of onset in these groups has not been examined. The purpose of this study was to compare age of first hospitalization of (1) native-born people versus immigrants, (2) immigrants from different countries of origin, and (3) first generation immigrants versus second generation immigrants; and to reexamine gender differences in age of first hospitalization. Data were extracted on all first hospital admissions nationally for the years 1978-1992 (n = 10,902) from the National Psychiatric Hospitalization Case Registry of the State of Israel Ministry of Health. Immigrants were older at time of first hospitalization than nonimmigrants, with considerable variations between different countries of origin. Second generation immigrants (i.e., born in Israel to immigrant parents) had ages of first hospitalization similar to people with native-born parents. Males had earlier ages at first hospitalization than females. The results suggest that immigration may have a delaying effect on age of first admission and support previous findings regarding gender difference in age of onset. PMID- 12645682 TI - Clinical outcome of patients with schizophrenia without maintenance treatment in a nonindustrialized society. AB - The outcome of 51 patients with schizophrenia (DSM-III-R) consecutively admitted to Bangli State Mental Hospital (Bali) with no prior psychiatric treatment was assessed by the Positive and Negative Syndrome Scale (PANSS) and Eguma's Social Adjustment Scale (ESAS) at a 5-year followup evaluation. The subjects were divided into two groups according to their medication patterns determined at the assessment. Mann-Whitney U test revealed no significant differences in the PANSS scores between the treated group (n = 22) and the nontreated group (n = 29), whereas a significant difference in the distribution of the scores was observed (Kolomorov-Smirnoff test, p < 0.05). Subjects in the nontreated group showed a greater tendency to be scored either high or low in the PANSS than did subjects in the treated group. Moreover, subjects in the nontreated group tended to be classified more often into the best or worst outcome categories when the ESAS was performed than subjects in the treated group (chi-square analysis, p < 0.05). In the present study, nontreatment may reflect a good outcome in terms of no requirement for continuing treatment but still be causally related to poor outcome in terms of absence of the necessary treatment. The results suggest the importance of therapeutic intervention for schizophrenia patients who have severe psychopathology without maintenance treatment in nonindustrialized societies. PMID- 12645681 TI - Prefrontal structural and functional deficits in schizotypal personality disorder. AB - Structural prefrontal deficits have been reported in patients with schizophrenia, but it is unclear if they are also found in patients with schizophrenia spectrum personality disorders. The hypothesis that a spectrum group will be characterized by prefrontal structural deficits was tested by assessing prefrontal gray and white volumes using magnetic resonance imaging in a community sample of 16 individuals with schizotypal/paranoid personality disorder, 27 comparisons, and 26 psychiatric controls. Frontal neurocognitive functioning was also assessed using the Wisconsin Card Sorting Test and the Continuous Performance Test. The spectrum group showed reduced prefrontal gray volumes and poorer frontal functioning compared to both other groups. Structural deficits were independent of functional deficits and together correctly classified 84.2 percent of subjects. Structural but not functional deficits were abolished after a strict control for antisocial personality was made. Results support the notion that frontal deficits may be centrally involved in the etiology of schizophrenia but also suggest that comorbid antisocial behavior may be one factor accounting for differences in prefrontal structural findings across studies. PMID- 12645684 TI - At issue: management of medication noncompliance in schizophrenia by families in India. AB - Noncompliance with medication during a symptomatic phase is a common problem in the treatment of schizophrenia patients. In India, a majority of patients live with their families and those families supervise patients' medication intake. In a study of patients attending an urban outpatient care center in India, it was noted that when the patients were acutely ill and refused to take medication, the families administered medication to them without patients' knowledge, under the supervision of the psychiatrist. This method had been practiced by families in half the cases of patient noncompliance. Many families felt that there was no other viable alternative under the circumstances. Only a minority of patients was aware of having received medication through this method, and many of them reacted negatively to it. However, the patients were subsequently taking treatment voluntarily following the reduction in the severity of the behavioral disorder with the involuntary treatment. The issues involved in this form of treatment are discussed with regard to the social and health care environment in the country. PMID- 12645683 TI - Developing a partnership with NAMI and psychiatric pharmacists. AB - NAMI is a grassroots advocacy organization dedicated to improving the lives of people with severe mental illness. We conducted a survey of 8 local Southern California NAMI chapters and found the membership to be predominantly female (70%) and elderly (mean age = 61 years). The majority of respondents (92%) rated information on psychiatric medications as very important, and 79 percent reported needing more information on psychiatric medications. A second survey of psychiatric pharmacists nationwide revealed that 80 percent have heard of NAMI but only 17 percent are members of NAMI. Psychiatric pharmacists have expertise in psychopharmacology, and more than 50 percent have achieved board certification in psychiatric pharmacy (BCPP). A partnership between NAMI and psychiatric pharmacists can result in education of NAMI members about psychiatric medications. Furthermore, psychiatric pharmacists can become aware of issues that are pertinent to the experience of NAMI members. PMID- 12645685 TI - Concealed medicines for people with schizophrenia: a U.S. perspective. PMID- 12645687 TI - First person account: eliminating delusions. PMID- 12645686 TI - Consensus on early intervention in schizophrenia. PMID- 12645688 TI - Understanding logistic regression analysis in clinical reports: an introduction. PMID- 12645689 TI - Secondary prevention after coronary bypass: the American Heart Association "Get with the Guidelines" program. PMID- 12645690 TI - Thoracic Surgery Directors Association Award. Bone marrow as a cell source for tissue engineering heart valves. AB - BACKGROUND: This study was designed to assess the feasibility of using ovine bone marrow-derived mesenchymal stem cells to develop a trileaflet heart valve using a tissue engineering approach. METHODS: Bone marrow was aspirated from the sternum of adult sheep. Cells were isolated using a Ficoll gradient, cultured, and characterized based on immunofluorescent staining and the ability to differentiate down a specific cell lineage. Two million cells per centimeter squared were delivered onto a polyglycolic acid (PGA), poly-4-hydroxybutyrate (P4HB) composite scaffold and cultured for 1 week before being transferred to a pulse duplicator for an additional 2 weeks. The tissue-engineered valves were assessed by histology, scanning electron microscopy, and biomechanical flexure testing. RESULTS: Cells expressed SH2, a marker for mesenchymal stem cells, as well as specific markers of smooth muscle cell lineage including alpha-smooth muscle actin, desmin, and calponin. These cells could be induced to differentiate down an adipocyte lineage confirming they had not fully committed to a specific cell lineage. Preliminary histologic examination showed patchy surface confluency confirmed by scanning electron microscopy, and deep cellular material. Biomechanical flexure testing of the leaflets showed an effective stiffness comparable to normal valve leaflets. CONCLUSIONS: Mesenchymal stem cells can be isolated noninvasively from the sternum of sheep and can adhere to and populate a PGA/P4HB composite scaffold to form "tissue" that has biomechanical properties similar to native heart valve leaflets. Thus, bone marrow may be a potential source of cells for tissue engineering trileaflet heart valves, particularly in children with congenital heart disease. PMID- 12645691 TI - Improvement of cardiac function by bone marrow cell implantation in a rat hypoperfusion heart model. AB - BACKGROUND: Local bone marrow cell implantation can induce angiogenesis. In the present study we investigated whether angiogenesis induced by bone marrow cell implantation improves deteriorated cardiac function in a rat heart model of hypoperfusion. METHODS: A hypoperfusion heart model was created in Dark Agouti rats by ligating the left anterior descending artery placed against a copper wire (phi275 microm), then pulling out the wire immediately. The left ventricular (LV) anterior wall was injected directly at six points, each with 1 x 10(7) bone marrow cells in 10 microL of phosphate-buffered saline or with phosphate-buffered saline only, respectively. Echocardiography was performed to evaluate the cardiac function 7, 30, 60, and 90 days after treatment. Microvessel density and blood flow in the LV anterior wall were estimated 60 days after treatment. RESULTS: Both the increase of LV end-systolic diameter and the decrease of percent of fractional shortening caused by myocardial ischemia were attenuated effectively by bone marrow cell implantation treatment. Bone marrow cell implantation treatment also increased the levels of angiopoietin-1 and vascular endothelial growth factor in the LV anterior wall. The microvessel density, blood flow, and thickness of the LV anterior wall significantly also increased after bone marrow cell implantation treatment compared with those after phosphate-buffered saline injection. CONCLUSIONS: The local implantation of autologous bone marrow cells induced angiogenesis and improved the perfusion of ischemic myocardium, thereby preventing LV remodeling and improving deteriorated cardiac function caused by myocardial hypoperfusion. PMID- 12645692 TI - Transformation of adult mesenchymal stem cells isolated from the fatty tissue into cardiomyocytes. AB - BACKGROUND: Myocardial infarction results in the death of cardiomyocytes, which are replaced by scar tissue. Cardiomyocytes cannot regenerate because they are terminally differentiated. Mesenchymal cells are pluripotent cells, which have the potential to differentiate to specialized tissues under appropriate stimuli. The aim of this study was to direct differentiation of the adult mesenchymal stem cells isolated from fatty tissue into cardiomyocytes using 5-azacytidine. METHODS: Adult mesenchymal stem cells were isolated from the fatty tissue of New Zealand White rabbits and cultured in RPMI medium. Second-passaged mesenchymal cells were treated with various concentrations of 5-azacytidine and incubated for different intervals of time. The cells were plated in six-well dishes at 500, 5,000, and 50,000 cells/well. These cells were treated with 1-, 3-, 6-, 9-, and 12-micromol/L concentrations of 5-azacytidine and incubated for 12, 24, 48, and 72 hours. Later, the medium was replaced with fresh medium and incubated in a CO2 incubator. The medium was changed once at 3 to 4 days. At 2 months, the cells were fixed with 0.4% glutaraldehyde for 2 hours and later washed with phosphate buffered saline. The transformed cells were subjected to immunostaining for the myosin heavy chain, alpha actinin, and troponin-I. RESULTS: After treatment with 5-azacytidine, the adult mesenchymal stem cells were transformed into cardiomyocytes. At 1 week, some cells showed binucleation and extended cytoplasmic processes with adjacent cells. At 2 weeks, 20% to 30% of the cells increased in size and formed a ball-like appearance. At 3 weeks, these cells began to beat spontaneously in culture when observed under phase contrast microscope. Immunostaining of the transformed cells for myosin heavy chain, alpha actinin, and troponin-I was positive. The differentiated cells maintained the phenotype and did not dedifferentiate up to 2 months after treatment with 5 azacytidine. CONCLUSIONS: These observations confirm that adult mesenchymal stem cells isolated from fatty tissue can be chemically transformed into cardiomyocytes. This can potentially be a source of autologous cells for myocardial repair. PMID- 12645693 TI - Outpatients on mechanical circulatory support. AB - BACKGROUND: As waiting periods for heart transplantation have lengthened, the application of long-term mechanical circulatory support (MCS) has become more common in patients presenting with cardiogenic shock. Anticipating increased long term MCS, a policy to discharge patients home has been instituted. This study compares the results of outpatient on MCS to a group of patients remaining hospitalized. METHODS: We report our 10-year experience with 108 patients on MCS, who were supported for more than 3 months. Group A consisted of 38 patients (25 Novacor, 13 Berlin Heart) who underwent assist implantation from 1996 to 2001. They had a mean support time of 454 days (range 100 to 1074 days) and spent a mean of 326 days (range 20 to 769 days) at home. Group B consisted of 70 patients (24 Novacor, 46 Berlin Heart) who underwent assist implantation between 1991 and 2000. They had a mean support time of 234 days (range 95 to 795 days) and were not discharged. The patients were monitored for complications, hospital readmissions, and causes of death including infections and thromboembolic and bleeding events during the MCS time. RESULTS: Group A total mortality was 16% (6/38). Two patients died from cerebral embolism, one from cerebral hemorrhage, two from systemic infection, and one from multiorgan failure. Thirty-two patients (84%) required 95 readmissions to the hospital due to cerebral embolism (n = 9), bleeding (n = 1), wound infections (n = 23), coagulation disorder (n = 13) for heart transplantation (n = 5), and (n = 44). In group B the mortality was 43% (30/70) for noncardiac reasons and thus significant higher (p = 0.004, chi2 test). Causes of death were cerebral embolism (n = 5), cerebral hemorrhage (n = 7), systemic infection (n = 14, significantly higher, p = 0.04, chi2 test), and multiorgan failure (n = 4). CONCLUSIONS: Our experience demonstrates that MCS can be used in outpatients without increased mortality and with an acceptable rate of readmissions (2.8/patient). It ensures the survival of the patient, enables recovery from multiorgan failure, and offers an acceptable quality of life. PMID- 12645694 TI - Left ventricular aneurysm resection with port-access surgery: a new mini-invasive surgical approach. AB - BACKGROUND: In recent years port-access and endovascular extra-corporeal circulation techniques have allowed valvular and coronary operations to be performed by mini-thoracotomy. Experience with the technique suggested application to resection of ventricular aneurysms, which are usually approached through a median sternotomy with the use of traditional cardiopulmonary bypass. METHODS: We performed a left port-access mini-thoracotomy, with 6 to 8 cm skin incisions, in 7 patients undergoing endoventricular pericardial patch repair for anterior left ventricular aneurysm. Cardiopulmonary bypass was effected using the Heartport system. The mean interval between myocardial infarction and operation was 60.4 +/- 57.7 months. Three patients developed sustained ventricular tachicardia. Mean preoperative ejection fraction was 34% +/- 11%. Associated procedures were coronary bypass grafting in 2 patients and cryosurgery in 3 patients. RESULTS: All patients survived to discharge and are alive and well after an average 14.5 months. They are all in NYHA class I-II. Postoperative echocardiograms revealed an average ejection fraction of 48.0% +/- 7.5% (p = 0.006 compared with preoperative value). The 3 patients who had cryosurgery did not demonstrate any recurrence of arrhythmias. CONCLUSIONS: Left ventricular aneurysm can be successfully treated through port-access mini-thoracotomy with endovascular cardiopulmonary bypass, avoiding median sternotomy. This mini invasive approach allows effective ventricular remodeling. Revascularization and antiarrhythmia surgery can also be done at the same time. In case of severely reduced ventricular function this approach permits fibrillatory arrest without aortic cross-clamping. The results are also good in terms of hospitalization time and long-term survival. PMID- 12645695 TI - Improved neurohormonal markers of ventricular function after restoring sinus rhythm by the Maze procedure. AB - BACKGROUND: Clinical results of the Maze procedure for treatment of atrial fibrillation (AF) are excellent, suggesting improved ventricular function after restoring sinus rhythm. However, long-term corresponding effects on the release of cardiac natriuretic peptides and other vasoactive hormones are incompletely investigated after isolated Maze surgery. METHODS: Plasma levels of brain natriuretic peptide (BNP), atrial natriuretic peptide (ANP), antidiuretic hormone, aldosterone, and angiotensin II were measured in 15 patients (mean age, 52 +/- 11 years) undergoing isolated surgical Maze (III) procedures for medically refractory AF, preoperatively and 6 months postoperatively. At the time of blood sampling, hemodynamic correlates were obtained at baseline and after 6 and 12 minutes of rapid ventricular pacing at 150 stimulations/minute. RESULTS: All patients were free of AF at 6-month follow-up. The measured plasma levels of BNP, ANP, and angiotensin II were all significantly lower (p = 0.03) late after the isolated Maze procedure. Cardiac output was significantly higher postoperatively (p < 0.01). Other hemodynamic values and left atrial size were unchanged after surgery. Ventricular pacing caused almost identical hemodynamic changes in atrial pressures before and late after surgery, but the associated plasma ANP response was significantly attenuated postoperatively (p < 0.001). CONCLUSIONS: Levels of cardiac natriuretic peptides and angiotensin II as markers of ventricular function are improved in the long term after clinically successful isolated Maze procedures. ANP response to hemodynamic challenge by ventricular pacing was attenuated postoperatively, possibly due to atrial scarring. PMID- 12645696 TI - Older patients fare better with the Ross operation. AB - BACKGROUND: The Ross operation has an established position in young patients. We address the question of whether any age group profits most from the Ross operation, and we compare the results in various ages. METHODS: From February 1995 to August 2001 we performed 250 Ross operations. Group 1 consisted of 46 patients, ages 2 to 25 years (median age, 15 years). Group 2 consisted of 123 patients, ages 26 to 49 years (median age, 39 years). Group 3 consisted of 81 patients, ages 50 to 67 years (median age, 55 years). Echocardiography was performed perioperatively, at 2 to 6 months, and then yearly. RESULTS: Mean follow-up for the three groups was 32, 31, and 28 months, respectively (p = 0.36). One patient from group 2 died after 25 months caused by suppurative pneumonia and 3 patients from group 3 died (1 from suspected acute thoracic aorta dissection at 40 months, 1 from ventricular fibrillation after 25 months, and 1 from an undiagnosed sudden death at 5 months). Autograft replacement was necessary for 3 patients from group 2 and 1 from group 3. Autograft repair was necessary for 1 patient from group 2, and pulmonary homograft reoperation was necessary for 1 patient from group 1. All other autografts currently have physiologic gradients and clinically insignificant regurgitation. Median peak gradient across the right ventricular outflow tract was 23.6 +/- 18 mm Hg for group 1, 14.6 +/- 8 mm Hg for group 2, and 11.5 +/- 7 mm Hg, which was significantly lower for group 3 patients (p < 0.001). Eleven patients are under close follow-up for right ventricular outflow tract gradients > or = 40 mm Hg; eight of these patients are from group 1, 3 are from group 2, and there are none from group 3. CONCLUSIONS: Although the Ross operation provides excellent results in all age groups, the problem of right ventricular outflow tract stenosis has not been seen in patients older than 50 years, which implies that it offers superior results for aortic valve disease in middle aged and older patients. PMID- 12645698 TI - Papillary muscle sling: a new functional approach to mitral repair in patients with ischemic left ventricular dysfunction and functional mitral regurgitation. AB - BACKGROUND: In patients with ischemic left ventricular dysfunction (LVD) and functional mitral regurgitation (FMR), restoring a more normal alignment between mitral annulus and laterally displaced papillary muscles (PM) may be beneficial in terms of mitral repair and regional dynamics. METHODS: Ten patients, 29 to 78 years old, with an ejection fraction of 25% to 45%, pulmonary hypertension greater than 60, and New York Heart Association Class III-IV, had their PMs drawn together by a tightly encircling loop using a 4-mm Gore-Tex tube. Associated mitral annuloplasty rings were only moderately undersized. Efficiency was essentially evaluated on reversal of mitral tenting and control of FMR. RESULTS: Postoperative echocardioraphy revealed changes in "tenting effect" from 14 +/- 2.8 mm to 4 +/- 1.41 mm. Regurgitation is none to trivial in 9 patients, and mild in 1 patient. The posterior left ventricular wall between the PMs is shortened as a result of the surgical remodeling and may be beneficial on local dynamics. CONCLUSIONS: Joining the PM side-by-side has an obvious immediate effect on mitral leaflet mobility by suppressing the tethering due to displacement of the PM. An eventual result on local ventricular dynamics needs confirmation. PMID- 12645697 TI - Stentless aortic valves as an alternative to homografts for valve replacement in active infective endocarditis complicated by ring abscess. AB - BACKGROUND: The valve substitute of choice in active infective aortic valve endocarditis complicated by annulus abscess in our institution is the cryopreserved homograft. To avoid implantation of any prosthetic material, the Shelhigh No-React stentless valves and conduits may be considered an alternative when no suitable homograft is available. METHODS: Between March 1986 and January 2001, 452 homografts were implanted in the aortic position. From January 2000 to August 2001, 75 Shelhigh No-React prostheses were implanted at our institution. In 25 consecutive patients (study group) with aortic annulus abscess, urgent aortic valve replacement with the Shelhigh SuperStentless and Stentless Aortic Valve Conduit was undertaken. Patients (16 male, 9 female; age, 49 +/- 19 years) were studied with follow-up until March 2002. The control group comprised 68 consecutive historical patients (46 male, 22 female; age, 53 +/- 14.4 years) with similar disease treated between January 1997 and December 1999 in whom an aortic homograft was implanted. This group was also followed up until March 2002. Demographic data and preoperative characteristics of the patients were without significant differences. Patients were studied by echocardiography. RESULTS: Sixty-day mortality was 16% (11 patients) in the control group compared with 12% (3 patients) in the study group. Recurrent infection occurred in 4% in both groups. The instantaneous and mean Doppler gradients yielded no significant differences (19.4 +/- 10.4 mm Hg and 11.8 +/- 5.7 mm Hg versus 18.2 +/- 8.7 mm Hg and 10.9 +/- 5.3 mm Hg, respectively). The mean effective orifice area calculated from Doppler flow velocity for the stentless valve was 2.3 +/- 0.6 cm2. Preoperative evaluation of left ventricular dimensions and global left ventricular systolic function did not vary significantly between the two groups. However, postoperatively evaluated left ventricular end-diastolic diameter dimensions in the study group were significantly smaller than those in the control group (47.6 +/- 7.9 mm versus 56 +/- 9.5 mm; p = 0.05). Ejection fraction was similar in both groups (56.2% +/- 12.8% for the study [Shelhigh] and 52.6% +/ 16.8% for the control [homograft] group). CONCLUSIONS: Our experience with both the Shelhigh No-React SuperStentless and Stentless Aortic Valve Conduit in patients with native or prosthetic aortic valve endocarditis appears to demonstrate good results, similar to those of cryopreserved homografts. Ease of implantation and favorable effective orifice area and pressure gradients, as well as the No-React anticalcification treatment, are promising factors. PMID- 12645699 TI - Mitral valve surgery and acute renal injury: port access versus median sternotomy. AB - BACKGROUND: Many outcomes and complications of minimally invasive and conventional cardiac surgery await comparison. Patients undergoing mitral valve surgery commonly sustain renal injury. Using peak postoperative fractional change of serum creatinine as a marker of renal injury, we tested the hypothesis that mitral valve surgery with port access minithoracotomy (Port) and conventional surgery with a median sternotomy (MS) incision are associated with different degrees of acute renal injury. METHODS: We evaluated data from all isolated mitral valve operations by a single surgeon between 1990 and 2000 (MS = 90, Port = 227). We also performed a secondary analysis of mitral valve surgeries performed by both MS and Port approaches in a concurrent period from 1996 to 2002 (MS = 93, Port = 240). Univariable and multivariable tests were used to determine the association of surgical technique with peak postoperative creatinine (CrmaxPost) and peak postoperative fractional change in creatinine (%deltaCr); p less than 0.05 was considered significant. RESULTS: In our analysis that accounted for the date of surgery, we observed a highly significant independent association between surgical approach and %deltaCr, indicating a greater risk of acute renal injury in the MS group (F value 13.33; p = 0.0003). Similar findings were noted in the secondary (time-concurrent) analysis of %deltaCr (F value 12.65; p = 0.0176). CONCLUSIONS: We present retrospective evidence of reduced acute renal injury associated with the port access technique in mitral valve surgery patients. Our findings suggest that a port access minithoracotomy approach to mitral valve surgery may be preferable to conventional methods for patients with high renal risk. PMID- 12645700 TI - Use of mitral valve repair: analysis of contemporary United States experience reported to the Society of Thoracic Surgeons National Cardiac Database. AB - BACKGROUND: The Society of Thoracic Surgeons National Cardiac Database data indicate that the performance of mitral repair has increased significantly (1990 to 23.2%, 1999 to 32.0%, p < 0.0001). We examined contemporary (1999 to 2000) usage of mitral repair in the United States. METHODS: We analyzed National Cardiac Database data to determine the operative approach for a diagnosis of mitral regurgitation. RESULTS: A total of 21,741 isolated and combined mitral valve procedures were identified. The overall frequency of repair was 37.7% (8206). For procedures isolated to the mitral valve, the frequency of repair was 35.7% (3027/8486) whereas repair was more common with concomitant CABG (42.9% [3088/7193], p < 0.0001). The proportion of patients having repair decreased with age (41.2% [386/936] in 20 to 39 years, 36.1% [3513/9746] in > 70 years, p = 0.0016). Repair was more common in males (43.5% [4720/10860]) than females (32.0% [3472/10842], p < 0.0001). Repair was less common as NYHA Class increased (Class I, 47.8% [949/1984] vs Class IV, 33.2% [1803/5427]) and for emergent operative status (21.2% [156/736] vs 38.5% [8000/20773] for elective/urgent, both p < 0.0001). The number of prior operations did not affect the use of repair. Simple annuloplasty was performed in the majority of reported repairs (62.8% [3837/6115]), more so with associated CABG as compared to isolated repair (70.2% [2167/3088] vs 55.2% [1670/3027]; p < 0.0001). CONCLUSIONS: Mitral repair was performed in over one-third of the patients reported in 1999 to 2000 and has increased since the National Cardiac Database inception. Repair usage differed based on sex, age, gravity of illness, and associated procedures. This provides a base line from which to expand the application of repair. PMID- 12645701 TI - Long-term results after Starr-Edwards mitral valve replacement in children aged 5 years or younger. AB - BACKGROUND: Mitral valve replacement with prosthetic valves in infants and children requires consideration of multiple variables. When we examined our late follow-up, the effect of the growth of the patient relative to the size of the prosthesis placed was the most important variable predicting late successful results. We reviewed our experience with mitral valve replacement using the Starr Edwards ball valve in children aged 5 years or younger, focusing on the effect of valve prosthesis-patient mismatch on the long-term results in the growing patient. METHODS: From August 1974 to June 1986, 8 patients aged 5 years or younger underwent mitral valve replacements using the Starr-Edwards prosthesis size OM in 3 patients and 1M in 5 patients. Model 6320 was used in 1 patient and Model 6120 in the remaining 7 patients. RESULTS: Follow-up was 100% from 15 to 27 years (mean, 20 years). No valve-related complications of thromboembolism, anticoagulant-related hemorrhage, or prosthetic valve endocarditis were seen. All patients normally developed to adult size. The range of the valve area index of the 3 patients who received the smaller Starr-Edwards valve (size OM) was 0.97 to 1.24 cm2/m2. Although this size valve was adequate for patient growth to adolescence, in each case valve replacement with a larger valve was required. CONCLUSIONS: Our long-term review of Starr-Edwards ball valve mitral valve replacement in children aged 5 years or younger shows that the Starr-Edwards ball valve (Models 6320 [1 patient] and 6120 [7 patients]) showed excellent durability, no thromboembolism, and no anticoagulant-related complications. Size OM valves required replacement for hemodynamic reasons because of patient growth; larger size 1M valves remained hemodynamically satisfactory in spite of patient growth. PMID- 12645702 TI - Outcomes in nonagenarians after heart valve replacement operation. AB - BACKGROUND: Changes in the age profile of the United Kingdom population and improvements in preoperative and postoperative care have resulted in increasing numbers of very elderly patients undergoing heart valve replacement (HVR) operations. Although HVR operations in nonagenarians are relatively uncommon, the demand for cardiac operations in this age group may increase over time. Outcomes after HVR operations in nonagenarians have not been well described yet. Therefore, the aim of this study was to determine outcomes in terms of early mortality and long-term survival in 35 nonagenarians after HVR operation. METHODS: Data from the United Kingdom Heart Valve Registry were analyzed and nonagenarian patients were identified. Additional analyzed data include gender, valve position, valve type, valve size, operative priority, follow-up time, and date and cause of death. Kaplan-Meier actuarial curves were calculated to determine accurate 30-day mortality and long-term survival. RESULTS: On average five HVR operations are performed annually in the United Kingdom in nonagenarians with equal numbers of males and females. Aortic valve replacement with a bioprosthetic valve was the most common operation and 86% were elective admissions. Fourteen patients died within the review period; mean time to death was 402 days. Overall 30-day mortality was 17%, which was higher for males compared with females; females also displayed better long-term survival. CONCLUSIONS: HVR operations in nonagenarians carry a significantly higher risk of early mortality and reduced long-term survival. Despite increases in the age profile of the population, elective HVR operation with patients aged 90 years or older is likely to remain an infrequent surgical procedure reserved for very carefully selected patients. PMID- 12645703 TI - Cognitive self-assessment one year after on-pump and off-pump coronary artery bypass grafting. AB - BACKGROUND: Coronary artery bypass grafting (CABG) is associated with significant cerebral morbidity. This is usually manifested as cognitive decline and may be caused by cardiopulmonary bypass. The primary objective of this study was to explore whether patients report more cognitive failures 1 year after CABG than preoperatively. Secondary objectives were to evaluate whether there is a difference in reported cognitive failures between patients undergoing on-pump and off-pump CABG and whether a difference between CABG patients and healthy control subjects exists. Finally the relation between objective and subjective cognitive functioning was quantified. METHODS: In this prospective study, the Cognitive Failures Questionnaire (CFQ) was assigned preoperatively and 1 year postoperatively to 81 patients who were randomly assigned to undergo off-pump (n = 45) or on-pump (n = 36) CABG. A control sample of 112 age-matched healthy subjects was included who were administered the CFQ once. RESULTS: No difference was found in the total CFQ score (p = 0.222) and CFQ worry score (p = 0.207) between 1 year after CABG and before CABG. There was no difference between on pump and off-pump CABG (total score, p = 0.458; worry score, p = 0.563). A significant difference was found in CFQ total score between CABG patients and control subjects (p < 0.001), with control subjects reporting more cognitive failures than CABG patients. Finally, patients who showed cognitive decline in the Octopus trial did not have a higher CFQ total score (p = 0.671) and CFQ worry score (p = 0.772) than patients without cognitive decline 1 year after CABG. CONCLUSIONS: The present findings suggest that CABG does not result in a substantial proportion of patients with subjectively experienced cognitive decline 1 year after the procedure, irrespective of the type of surgical technique (on-pump versus off-pump). PMID- 12645704 TI - Comparative effects of propofol versus fentanyl on cerebral oxygenation state during normothermic cardiopulmonary bypass and postoperative cognitive dysfunction. AB - BACKGROUND: The purpose of this study was to examine the comparative effects of propofol and fentanyl on cerebral oxygenation during normothermic cardiopulmonary bypass and postoperative cognitive dysfunction. METHODS: One hundred eighty patients scheduled for elective coronary artery bypass grafting were randomly divided into two groups: propofol group (n = 90) and fentanyl group (n = 90). After induction of anesthesia, a fiberoptic oximetry oxygen saturation catheter was inserted into the right jugular bulb to monitor jugular venous oxygen hemoglobin saturation continuously. Hemodynamic measurements and arterial and jugular venous blood gases were measured at seven time points. All patients underwent a battery of neurologic and neuropsychological tests on the day before the operation and at 6 months after the operation. RESULTS: Cerebral desaturation (defined as a jugular venous oxygen hemoglobin saturation value less than 50%) during cardiopulmonary bypass was more frequent in the fentanyl group than in the propofol group. Cerebral desaturation time (duration when jugular venous oxygen hemoglobin saturation was less than 50%) and the ratio of cerebral desaturation time to total cardiopulmonary bypass time in the fentanyl group differed significantly from those in the propofol group (fentanyl group: 27 +/- 14 minutes, 20% +/- 9%; propofol group: 18 +/- 11 minutes, 14% +/- 7%, respectively, p < 0.05). There was no significant difference in postoperative cognitive dysfunction at 6 months after operation between the two groups (propofol group: 5 of 77, 6%; fentanyl group: 5 of 75, 7%). CONCLUSIONS: Propofol preserved cerebral oxygenation state estimated by jugular venous oxygenation during cardiopulmonary bypass compared with the fentanyl group. However, propofol did not affect postoperative cognitive dysfunction. PMID- 12645706 TI - Intraoperative and postoperative risk factors for respiratory failure after coronary bypass. AB - BACKGROUND: Unlike preoperative events, the influence of intraoperative or postoperative events on respiratory failure after coronary artery bypass grafting (CABG) remains unclear. The purpose of this study was to identify intraoperative and postoperative risk factors that predispose respiratory impairment after CABG. METHODS: A single institutional database combined with a mandatory report submitted to the Cardiac Surgery Registry of the New York State Department of Health was used. A total of 8,802 consecutive patients who underwent primary CABG with or without a concomitant cardiac operation from January 1993 through December 2000 were included. Respiratory failure was defined as the need for postoperative mechanical ventilatory support longer than 72 hours. Univariate and multivariate logistic regression model was used in the analysis. RESULTS: Of 8,802 consecutive patients (6,234 males and 2,568 females) who underwent CABG with or without a concomitant operation, 491 patients (5.6%) suffered from postoperative respiratory failure. Although univariate analysis identified 39 statistically significant preoperative risk factors for post-CABG respiratory failure, only six preoperative risk factors were statistically significant by multivariate analysis (p < 0.001). CPB time (in 30 minutes increments) was the only validated intraoperative variable that increased the risk of postrespiratory failure (odds ratio [OR], 1.2; p less than 0.0001). Postoperative events contributing significantly to an increased risk of post-CABG respiratory failure were (1) sepsis and endocarditis (OR, 90.4; p < 0.0001), (2) gastrointestinal bleeding with or without infarction and perforation (OR, 38.8; p < 0.0001), (3) renal failure (OR, 30.7; p < 0.0001), (4) deep sternal wound infection (OR, 11.3; p < 0.0001), (5) new stroke, intraoperative at 24 hours (OR, 9.3; p < 0.0001), and (6) bleeding that required reoperation (OR, 5.5; p < 0.0001). All perioperative variables together accounted for only 28.6% (R2) of the variation. CONCLUSIONS: Respiratory function after CABG is readily influenced by postoperative occurrence of extracardiac organ or system complications. PMID- 12645705 TI - Coronary reoperations: recurrence of angina and clinical outcome with and without cardiopulmonary bypass. AB - BACKGROUND: We compared our experience of minimal tissue dissection and target vessel revascularization without cardiopulmonary bypass (CPB) with the standard procedure of total dissection of the heart and complete revascularization with CPB in patients who had elective reoperative coronary artery bypass grafting (redo-CABG). METHODS: We analyzed recurrence of angina and clinical outcome in 118 patients who had elective redo-CABG between January 1995 and April 2002. Seventy-four patients had redo-CABG with CPB, and 44 patients had redo-CABG without CPB. RESULTS: Perioperative outcome was comparable with regard to morbidity and mortality rates. At follow-up, the mean Canadian Cardiovascular Society score was 1.3 +/- 0.6 in patients who had redo-CABG with CPB and 1.7 +/- 0.8 in patients who had redo-CABG without CPB (p = 0.02). At follow-up, patients who had redo-CABG without CPB had a higher rate of recurrence of angina (log rank = 0.001) and higher use of nitrates (p = 0.015). Target vessel revascularization was an independent predictor of recurrence of angina in younger patients (< 75 years; p = 0.012) but not in the elderly (> or = 75 years; p = 0.142). CONCLUSIONS: In elective redo-CABG patients, minimal tissue dissection and target vessel revascularization without cardiopulmonary bypass did not add significant benefit with regard to perioperative morbidity and mortality. The unsatisfactory relief of symptoms does not seem to justify target vessel revascularization by a less invasive approach. Therefore, this technique should be offered exclusively to patients at high risk with complete revascularization using CPB, such as the elderly. PMID- 12645708 TI - Coronary endarterectomy with off-pump coronary artery bypass surgery. AB - BACKGROUND: The aim of this study is to review our experience in coronary artery endarterectomy performed without cardiopulmonary bypass. METHODS: Between May 1998 and June 2000 off-pump coronary endarterectomy was performed on 11 patients who had unstable angina pectoris. The mean ejection fraction (EF) was 26.3 +/- 4.4, and all of the patients were New York Heart Association (NYHA) III or IV. Off-pump open left anterior descending (LAD) endarterectomy was performed on 7 patients, and closed endarterectomy of the right coronary artery (RCA) was done on the remaining 4. RESULTS: There were no deaths. None of the procedures was converted to on-pump operation; all the endarterectomies and bypasses were performed on the beating heart. All patients were completely revascularized, the left internal mammary artery was bypassed to the LAD in all operations, and all other grafts were of saphenous vein. At the end of the first year all bypasses to the endarterectomized arteries were patent. The overall patency rate was 95.6%. The mean postoperative EF was 34.7 +/- 9.1, which was significantly higher than the preoperative one (p < 0.05). At the end of the first year 9 patients were NYHA I or II and all were angina free in Canadian Cardiovascular Society class 0 or I. CONCLUSIONS: Endarterectomy without cardiopulmonary bypass can be performed in patients with severe left ventricular dysfunction who are expected to benefit from the complete revascularization. PMID- 12645707 TI - Prophylactic dialysis in patients with renal dysfunction undergoing on-pump coronary artery bypass surgery. AB - BACKGROUND: Preoperative creatinine values higher than 2.5 mg/dL are associated with markedly increased risk for both mortality and morbidity in patients undergoing coronary artery bypass surgery. We aimed to determine the effects of prophylactic perioperative hemodialysis on operative outcome in patients with nondialysis-dependent moderate renal dysfunction. METHODS: Forty-four adult patients with creatinine levels greater than 2.5 mg/dL but not requiring dialysis underwent coronary artery bypass surgery with cardiopulmonary bypass. The patients were randomly divided into two groups. In group 1 (dialysis group, 21 patients), perioperative prophylactic hemodialysis was performed in all patients. Group 2 (23 patients) was taken as a control group and hemodialysis was performed only if postoperative acute renal failure was diagnosed. RESULTS: The hospital mortality was 4.8% (1 patient) in the dialysis group, and 30.4% (7 patients) in the control group (p = 0.048). Postoperative acute renal failure requiring hemodialysis was seen in 1 patient (4.8%) in the dialysis group and in 8 patients (34.8%) in the control group (p = 0.023). Thirty-three postoperative complications were observed in the control group for an early morbidity of 52.2% (12 patients) and 13 complications occurred in 8 patients in the dialysis group (38.1%). The average length of the intensive care unit and postoperative hospital stay were shorter in the dialysis group than in the control group (p = 0.005 and p = 0.023, respectively). CONCLUSIONS: Preoperative creatinine levels higher than 2.5 mg/dL, increase the risk of mortality and the development of acute renal failure and prolong the length of hospital stay after on-pump coronary artery bypass surgery. Perioperative prophylactic hemodialysis decreases both operative mortality and morbidity in these high-risk patients. PMID- 12645709 TI - Preliminary experience with a novel intraoperative fluorescence imaging technique to evaluate the patency of bypass grafts in total arterial revascularization. AB - BACKGROUND: Early graft failure is a common cause of cardiac morbidity and mortality after coronary artery bypass grafting (CABG), and there is particular concern about graft patency in off-pump CABG. We describe our preliminary experience with a novel imaging technique (the SPY system), based on fluorescence of Indocyanine Green when exposed to near infrared light, for the intraoperative assessment of coronary graft patency. METHODS: Graft patency was assessed in patients undergoing off-pump and on-pump total arterial revascularization. The imaging technique requires injection of a 1-mL bolus of Indocyanine Green into the central venous line, followed by imaging with the SPY system. RESULTS: We assessed intraoperative graft patency in 213 conduits in 84 patients (mean, 2.54 grafts per patient), of which, 65 (77%) were done off-pump. It took approximately 3 minutes to image each graft. Skeletonized conduits provided better visualization than pedicled ones. Fluorescence, confirming graft patency, was observed in all but four (1.9%) conduits in 4 (5%) patients. In these latter cases, graft revision was necessitated. CONCLUSIONS: Fluorescence imaging of coronary grafts using the SPY is a uniquely simple, safe, noninvasive, and reproducible technique for intraoperative confirmation of graft patency. In 4 patients, it necessitated revision of the initial intraoperative procedure. Quantification of graft flow would enhance the value of the system. PMID- 12645710 TI - Endothelin receptors in endothelium-denuded human coronary artery bypass grafts and coronary arteries. AB - BACKGROUND: Coronary artery bypass graft (CABG) surgery is hampered by deleterious vasospasm in the vessel wall, especially in vein grafts. Endothelin (ET) is a strong vasoconstrictor that can be observed in increasing concentrations during CABG surgery. METHODS: Endothelin-induced vasoconstriction was evaluated in isolated, endothelium-denuded vessel segments of the human saphenous vein (SV), left internal mammary artery (LIMA), and coronary arteries. The ET(A) and ET(B) receptor mRNA levels were quantified by real-time polymerase chain reaction (PCR) analysis. RESULTS: The ET(A) and ET(B) receptor mRNA levels were significantly higher in the SV than in the LIMA and the coronary arteries. ET-1 induced a more efficacious contraction in the SV and LIMA as compared with in the coronary arteries. The ET(B) receptor agonist, Sarafotoxin 6c (S6c) stimulated constriction of the LIMA and SV, while inactive in the coronary arteries. The concentration-response curve for S6c was biphasic, suggesting activation of ET(A) receptors at high concentrations as this response could be inhibited by FR139317 (10 micromol/L), and ET(B) at low concentrations as this response could be inhibited by BQ788 (0.1 micromol/L). CONCLUSIONS: Endothelin induced vasoconstriction is mediated by ET(A) receptors alone in coronary arteries, while a combination of ET(A) and ET(B) receptors are of importance in SV and LIMA. Expression of contractile ET(B) receptors may be a pharmacologic disadvantage that contributes to the vasospasm during CABG surgery. The lower levels of ET(A) and ET(B) receptor mRNA in the LIMA and coronary arteries as compared with in the SV may provide one explanation for the better long- and short-term patency of LIMA as compared with SV grafts. PMID- 12645711 TI - The radial artery is larger than the ulnar. AB - BACKGROUND: The radial artery is presently widely used as a bypass graft for coronary artery reconstruction. However, the traditional opinion that the ulnar artery is the larger forearm artery has been questioned. METHODS: The internal diameters of the radial and ulnar arteries were measured at the wrist in postmortem angiograms of 24 cadavers. Differences in mean values of variables between ulnar and radial arteries were analyzed using the Wilcoxon test. RESULTS: The mean diameter of the radial artery was 28% larger than that of the ulnar artery in the right arm (p < 0.001) and 26% larger in the left arm (p < 0.001). In the right arm the radial artery was dominant in 20 of 24 cadavers (83%), the ulnar artery in 3 of 24 (13%), and the arteries were equal in 1 of 24 (4%). In the left arm the figures were 17 of 24 cadavers (71%), 3 of 24 (13%), and 4 of 24 (17%), respectively. CONCLUSIONS: In view of the present investigation the radial artery should be considered the larger forearm artery of the hand in most patients. PMID- 12645712 TI - In vivo validation of cardiac spiral computed tomography using retrospective gating. AB - BACKGROUND: Cardiac functional assessment represents the basis for diagnostics and cardiac operation planning. Spiral computed tomography (CT) combines the advantages of three-dimensional imaging and high temporal resolution when using gating techniques. However, in vivo validation data of this novel imaging technology are lacking. The purpose of this study was to validate in vivo the new imaging method using retrospective gating and to evaluate the clinical usefulness of the achieved temporal resolution. METHODS: In domestic pigs (n = 10, weight 35 to 40 kg) a flowmeter was placed surgically on the ascending aorta. Flow velocity integrated over systole served as the gold standard for left ventricular (LV) stroke volume (LVSV-FM). CT signal, projection data, pacemaker signal, and flow velocity were recorded simultaneously at constant heart rate (pacemaker, 90 beats per minute). End-systolic and end-diastolic frames were calculated by retrospective gating. LV volumes were traced, the difference representing CT stroke volume (LVSV-CT). Image data were three-dimensionally reconstructed using ray-tracing. RESULTS: Temporal resolution was 170 ms. Correlation of stroke volumes was high (r = 0.94, mean difference 1.75 mL). Intraobserver (0.49 mL for LVEDV, 0.31 for LVESV) and interobserver variability (p = 0.21 and p = 0.06, respectively) were low. Postprocessing resulted in four-dimensional beating-heart models useful for operation planning. CONCLUSIONS: Spiral CT using retrospective gating was validated in vivo. Clinically acceptable temporal resolution and accuracy in determining cardiac stroke volumes were found. As a true volumetric imaging modality the method may now play an important role in computer-assisted diagnostics and surgery. PMID- 12645714 TI - Neutrophil CD11b upregulation during cardiopulmonary bypass is associated with postoperative renal injury. AB - BACKGROUND: Renal injury remains a persistent complication of cardiopulmonary bypass (CPB) that, when sufficient to require dialysis, increases mortality eight fold. The high prevalence of renal failure in sepsis and adult respiratory distress syndrome has been linked to the systemic inflammatory response associated with those disorders. We hypothesized that components of the inflammatory response to CPB may similarly contribute to post-CPB acute renal injury. METHODS: Markers of leukocyte and platelet activation peri-CPB were measured in 75 patients undergoing cardiac operation with CPB and were correlated with acute renal injury, defined as an increase (> or = 50%) in peak serum creatinine post-CPB. RESULTS: Eleven patients sustained post-CPB acute renal injury. This subset of patients demonstrated significantly greater increases in neutrophil CD11b density (p = 0.01), as well as higher total neutrophil counts (p = 0.045), compared with patients with preserved renal function. Hemodynamic instability sufficient to require postoperative hemodynamic support also predicted an increased risk of acute renal injury. However, neutrophil CD11b upregulation did not correlate with this or any other clinical variables associated with renal risk, suggesting that this marker of the neutrophil inflammatory response may independently predict renal injury. By contrast other inflammatory markers, neutrophil myeloperoxidase levels, monocyte CD11b, base line C-reactive protein, and platelet CD62P expression did not differ between the two patient groups. CONCLUSIONS: Upregulation of the neutrophil adhesion receptor CD11b and high circulating neutrophil numbers are associated with acute renal injury after CPB, suggesting a contribution by activated neutrophils to the pathophysiology of this complication. PMID- 12645713 TI - Erythrocyte-containing versus crystalloid cardioplegia in the rat: effects on myocardial capillaries. AB - BACKGROUND: The purpose of this study was to investigate the effects of crystalloid and erythrocyte-containing cardioplegia on capillary morphology of the isolated erythrocyte-perfused rat heart. METHODS: Hearts from adult Sprague Dawley rats were perfused throughout with resuspended sheep erythrocytes and subjected to the following protocols (n = 6, all groups): (1) 15 minutes nonworking and 30 minutes working heart mode (control; group 1); (2) as for group 1, with 30 minutes erythrocyte-containing (BL) or crystalloid (CR) cardioplegic arrest without reperfusion (groups 2BL and 2CR); (3) as for group 2, with 30 minutes nonworking reperfusion (groups 3BL and 3CR); and (4) as for group 3, with 30 minutes working heart mode (groups 4BL and 4CR). After each protocol troponin I from coronary effluent was measured. Corrosion casts were then made of the coronary microvasculature. Cast density was calculated as cast volume per left ventricular dry weight. Casts also underwent scanning electron microscopy. Analysis was by analysis of variance. Values are mean +/- standard deviation. RESULTS: Prearrest working heart coronary flow averaged 15.1 +/- 4.7 mL/min without any differences among groups. Coronary flow in group 4 working hearts was the same before and after either cardioplegia. Cardiac outputs were similarly consistent in all groups. Cast density in group 1 (control) was 9.60 +/- 1.17 x 10(-2) mm3/mg. It was unaltered by erythrocyte-containing cardioplegia, but after crystalloid cardioplegia (group 2CR), it was 6.52 +/- 0.93 x 10(-2) mm3/mg (p = 0.0001 versus group 1 and p = 0.0007 versus group 2BL). With 30 minutes of nonworking reperfusion (group 3CR, there was slight improvement in cast density at 7.60 +/- 0.90 x 10(-2) mm3/mg (p = 0.0072 versus group 1; p = 0.0242 versus group 3BL). No further improvement was seen in group 4CR. Electron micrographs showed circumferential angularities or narrowings in crystalloid-perfused, arrested hearts, consistent with ischemic damage. Troponin I rose significantly after reperfusion in all groups, but it was higher in crystalloid-perfused, arrested hearts: 0.054 +/- 0.013 microg/L versus 0.024 +/- 0.017 microg/L (p = 0.0273). CONCLUSIONS: Erythrocyte-containing cardioplegia maintained capillary density and morphology. Crystalloid cardioplegia produced capillary loss, visible abnormalities, and higher troponin I release. These hearts may be more vulnerable to myocardial damage during reperfusion than hearts perfused with erythrocyte containing cardioplegic solution. PMID- 12645716 TI - Influence of PMEA-coated bypass circuits on perioperative inflammatory response. AB - BACKGROUND: Poly(2-methoxyethylacrylate) (PMEA) is a new coating material, and several experimental studies have revealed excellent biocompatibility of PMEA coated cardiopulmonary bypass circuits. The clinical utility of the PMEA-coated circuits was compared with that of uncoated circuits, focusing on perioperative inflammatory response. METHODS: Twenty-two patients were randomized to PMEA coated (group P; Capiox RX25; n = 11) or uncoated (group U; Capiox SX10; n = 11) circuit group, and underwent coronary artery bypass grafting and/or valve operations. The following markers, as well as clinical outcomes, were analyzed perioperatively: (a) complement activation by C3a (including C3a-desArg) concentrations; (b) leukocyte activation by polymorphonuclear-elastase concentrations; (c) acute phase inflammatory response by interleukin-6 concentrations; and (d) platelet preservation by number of platelets. RESULTS: The maximal values of C3a and polymorphonuclear-elastase were significantly lower in group P than in group U. The intergroup difference of interleukin-6 was not significant. Although preservation of platelets was significantly better in group P until 1 hour after initiating cardiopulmonary bypass, no significant intergroup difference was observed thereafter. The duration of postoperative mechanical ventilation revealed no significant intergroup difference. CONCLUSIONS: The PMEA coated circuits exhibited better suppression of perioperative complement and leukocyte activation than the uncoated circuits. In addition, the price of the PMEA-coated circuits is the same as that of the uncoated circuits. Therefore, we judged that the clinical utility of the PMEA-coated circuits is superior to those of the uncoated circuits. PMID- 12645715 TI - Is kidney function altered by the duration of cardiopulmonary bypass? AB - BACKGROUND: Cardiopulmonary bypass (CPB) is considered responsible for kidney damage. By using sensitive markers of kidney damage we assessed whether the length of CPB influences kidney function. METHODS: In a prospective study, 50 consecutive cardiac operation patients with CPB times of less than 70 minutes were compared with 50 consecutive patients showing CPB times of more than 90 minutes. Aside from creatinine clearance and fractional excretion of sodium, urine concentrations of N-acetyl-beta-D-glucosaminidase, alpha1-microglobulin, glutathione transferase-pi, and glutathione transferase-alpha were measured after induction of anesthesia at the end of the operation, and on the first and second postoperative days in the intensive care unit. RESULTS: CPB times were 58 +/- 12 minutes and 116 +/- 18 minutes, respectively. Hemodynamics, volume replacement, and use of catecholamines during cardiopulmonary bypass (CPB) were without significant differences between groups. Concentrations of all kidney-specific proteins increased significantly after CPB, showing the highest significant increases in the CPB more than 90 minutes group (eg, glutathione transferase alpha CPB > 90 minutes from 3.0 +/- 1.0 to 12.9 +/- 2.9 microg/L; glutathione transferase-alpha CPB < 70 minutes from 2.4 +/- 0.5 to 5.5 +/- 1.2 microg/L). By the second postoperative day, urine concentrations of kidney-specific proteins had returned to almost baseline in the CPB less than 70 minutes patients, but remained slightly elevated in the other group. CONCLUSIONS: Patients with CPB times more than 90 minutes showed more pronounced kidney damage than patients with CPB times less than 70 minutes as assessed by sensitive kidney-specific proteins. Whether patients with preexisting renal dysfunction undergoing prolonged CPB times would profit from renal protection strategies needs to be elucidated. PMID- 12645717 TI - Influence of two different perfusion systems on inflammatory response in pediatric heart surgery. AB - BACKGROUND: This study tests the hypothesis that a cardiopulmonary bypass system that combines complete heparin-coating, a centrifugal pump, and a closed circuit in comparison with a conventional system (uncoated system, roller pump, and hard shell venous reservoir) attenuates the inflammatory response in pediatric heart surgery. METHODS: In a prospective randomized controlled clinical study 40 consecutive children weighing 10 kg or less were included and divided into two groups. Concentrations of complement proteins (C3a, sC5b-9, C4d, and Bb), granulocyte degranulation products (polymorphonuclear [PMN] elastase), and proinflammatory cytokines (tumor necrosis factor [TNF]-alpha, interleukin [IL]-6, and IL-8) were measured. RESULTS: C3a and sC5b-9 concentrations were lower (C3a, p < 0.001; sC5b-9, p = 0.01) in the combined (heparin-coated/centrifugal pump/closed reservoir) group, the peak values being 58% and 37% of conventional group values. The Bb- and C4d-fragment values indicated activation of the complement system through the alternative pathway in both groups. PMN elastase concentrations were lower (p = 0.02) in the combined group, the peak values being 43% of conventional group values. There were no significant intergroup differences regarding TNF-alpha, IL-6, or IL-8 concentrations. CONCLUSIONS: The use of a fully heparin-coated system, a centrifugal pump, and a closed circuit during CPB in children (10 kg or less) leads to a lower degree of complement activation and PMN elastase release compared with a conventional system. PMID- 12645718 TI - A double-blind randomized trial: prophylactic vasopressin reduces hypotension after cardiopulmonary bypass. AB - BACKGROUND: Inhibition of angiotensin-converting enzyme (ACE) predisposes patients to vasodilatory hypotension after cardiopulmonary bypass (CPB). This hypotension has been correlated with arginine vasopressin deficiency and can be corrected by its replacement. In patients receiving ACE inhibition, we investigated whether initiation of vasopressin before CPB would diminish post-CPB hypotension and catecholamine use by avoiding vasopressin deficiency. METHODS: Cardiac surgical patients on ACE inhibitor therapy were randomized to receive vasopressin (0.03 U/min) (n = 13) or an equal volume of normal saline (n = 14) starting 20 minutes before CPB. RESULTS: Vasopressin did not change pre-CPB mean arterial pressure or pulmonary artery pressure. After CPB, the vasopressin group had a lower peak norepinephrine dose than the placebo group (4.6 +/- 2.5 versus 7.3 +/- 3.5 microg/min, p = 0.03), a shorter period on catecholamines (5 +/- 6 versus 11 +/- 7 hours, p = 0.03), fewer hypotensive episodes (1 +/- 1 versus 4 +/ 2, p < 0.01), and a shorter intensive care unit length of stay (1.2 +/- 0.4 versus 2.1 +/- 1.4 days, p = 0.03). CONCLUSIONS: In this cohort, prophylactic administration of vasopressin, at a dose without a vasopressor effect pre-CPB, reduced post-CPB hypotension and vasoconstrictor requirements, and was associated with a shorter intensive care unit stay. PMID- 12645719 TI - Cannulation of the axillary artery for cardiopulmonary bypass: safeguards and pitfalls. AB - BACKGROUND: The ascending aorta is the customary site for arterial cannulation for cardiopulmonary bypass. Favorable experience at our institution and elsewhere using axillary artery cannulation in treating type A aortic dissections has caused us to broaden our indications for using this site for arterial cannulation for cardiopulmonary bypass. METHODS: Medical records, operative notes, and perfusion records were reviewed in all patients in whom the axillary artery was cannulated directly or by a graft for cardiopulmonary bypass from January 1, 2000 through August 30, 2002. RESULTS: Seventy-five patients underwent axillary artery cannulation during the 32-month interval. Eleven patients had ascending aortic dissections, 20 had extensively diseased ascending aortas, and 44 were individuals undergoing repeat cardiac procedures. The right axillary artery was used in 72 patients and the left in 3. In 16 patients the artery was cannulated directly, and in 59 the arterial cannula was inserted into a prosthetic graft that had been anastomosed to the axillary artery. Axillary artery cannulation was satisfactory in 95% (71 of 75) of the cases in which it was used. CONCLUSIONS: Cannulation of the axillary artery for cardiopulmonary bypass is a dependable approach for procedures including reoperations, aortic dissections, and extensively diseased ascending aortas. PMID- 12645720 TI - Long-term results of cardiac and general health status in children after neonatal arterial switch operation. AB - BACKGROUND: The purpose of this study was to assess cardiac and general health status 8 to 14 years after neonatal arterial switch operation for transposition of the great arteries. METHODS: Sixty unselected children with intact ventricular septum (78.3%) or ventricular septal defect (21.7%) without or with aortic isthmic stenosis (5.1%) were examined 10.5 +/- 1.6 (mean +/- SD) years after neonatal switch and 5.3 +/- 1.6 years after mid-term evaluation. Complete clinical examination, standard and 24-hour Holter electrocardiogram, M-mode, 2D-, Doppler, and color Doppler echocardiography were performed. Results were compared with normal values and to mid-term follow-up results. RESULTS: Rates of reoperation after arterial switch operation and operation to correct concomitant coarctation were 3.3% and 5.1%, respectively. No patient needed medication, and 93.3% had no limitation of physical activity. All children had normal height and weight; 31.6% had abnormal thoracic configuration after median sternotomy. Most patients (91.7%) were in sinus rhythm. Incidence of complete right bundle branch block (10.0%) was unchanged, as was prevalence of ectopic activity (occasional atrial ectopy 20.0%, ventricular ectopy: occasional 21.7%; frequent 1.7%). Left ventricular dimensions and shortening fraction did not change over time. Diameters of neo-aortic valve annulus and neo-aortic root did not increase, and z scores decreased between mid-term and present evaluation. Incidence of neo-aortic insufficiency was 13.3% and remained unchanged in comparison with the pre examination value. Neo-aortic stenosis was not seen. Compared with mid-term follow-up, incidence (41.6%) and degree of supravalvular pulmonary stenosis increased. CONCLUSIONS: Good cardiac results persist 10 years after neonatal arterial switch operation for transposition of the great arteries. Encouraging findings include preservation of left ventricular function, low incidence of rhythm disturbances, lack of further neo-aortic root dilatation, and unchanged incidence of neo-aortic insufficiency compared with mid-term follow-up. Increased incidence and degree of supravalvular pulmonary stenosis are of concern. PMID- 12645721 TI - Detachment of the anterior leaflet of the tricuspid valve to expose perimembranous ventricular septal defects. AB - BACKGROUND: Detachment of the septal leaflet of the tricuspid valve has been described for better access to perimembranous ventricular septal defects. Detachment confined to the anterior leaflet is less known, although it provides a better exposure of the subaortic area and puts less jeopardy on the conduction tissues. METHODS: Data regarding 49 consecutive patients who had congenital perimembranous ventricular septal defect closure were retrospectively reviewed. Thirty-three patients (67%) underwent temporary detachment of the anterior leaflet of the tricuspid valve. The defect was closed with a Gore-Tex patch and a continuous suture. In 10 patients (29%), concomitant right ventricular outflow tract enlargement was performed. Follow-up was obtained in every patient (median time, 11 months; range, 2 to 26 months). RESULTS: No early or late death occurred. Closure of the ventricular septal defect was complete, with no more than trivial residual jet leaks found in perioperative echocardiography. All patients were in sinus rhythm. The tricuspid valve never showed more than mild insufficiency after repair. No patient showed subaortic obstruction. CONCLUSIONS: Detachment of the anterior leaflet of the tricuspid valve to expose the ventricular septal defect is a safe approach that allows rapid closure of the defect with a continuous suture and provides excellent results. PMID- 12645722 TI - Mesenteric blood flow response to feeding after systemic-to-pulmonary arterial shunt palliation. AB - BACKGROUND: We hypothesized that the splanchnic circulation protects against diastolic steal through a systemic-to-pulmonary arterial shunt by reducing its resistance. To test the hypothesis we compared the basal and postprandial mesenteric blood flow velocities and vascular resistance in infants after shunt palliation for their underlying cyanotic heart disease with those in nonshunted infants. METHODS: The basal and postprandial superior mesenteric arterial (SMA) time-average flow velocity (TAMV), end-diastolic flow velocity (EDFV), and relative resistance were assessed in 23 infants with congenital heart disease. The findings in the 9 shunted infants (group I) were compared with those in 14 nonshunted ones (group II). RESULTS: In group II, TAMV (0.25 +/- 0.07 versus 0.33 +/- 0.09 m/s, p < 0.001) and EDFV (0.08 +/- 0.04 versus 0.11 +/- 0.04 m/s, p = 0.003) increased, while SMA relative resistance decreased (297 +/- 121 versus 198 +/- 73 mm Hg/ms(-1), p < 0.001) postprandially. Similarly, in group I, TAMV (0.35 +/- 0.13 versus 0.48 +/- 0.19 m/s, p = 0.008) increased, while SMA relative resistance decreased (182 +/- 61 versus 116 +/- 38 mm Hg/ms(-1), p = 0.005) after feeding. However, whereas basal and postprandial diastolic flow was antegrade in group II, absent or retrograde diastolic flow was characteristic of group I (preprandial, -0.10 +/- 0.07 m/s; postprandial, -0.13 +/- 0.06 m/s). Furthermore, group I had significantly lower SMA relative resistance both before (p = 0.02) and after (p = 0.006) feeding. CONCLUSIONS: Profound disturbance of splanchnic perfusion occurs in infants palliated with a systemic-to-pulmonary arterial shunt. Their basal and postprandial SMA diastolic blood flow is either absent or reversed. The lowering of basal and postprandial resistance of the splanchnic circulation probably represents an adaptive mechanism to counteract such diastolic steal. PMID- 12645723 TI - A phase I study of Foscan-mediated photodynamic therapy and surgery in patients with mesothelioma. AB - BACKGROUND: Photodynamic therapy (PDT) is a light-based cancer treatment that, in the correct setting, can be delivered intraoperatively as an adjuvant therapy. A phase I clinical trial combining surgical debulking with Foscan-mediated PDT was performed in patients with malignant pleural mesothelioma. The purpose of the study was to define the toxicities and to determine the maximally tolerated dose (MTD) of Foscan-mediated PDT. METHODS: A total of 26 patients completed treatment. Tumor debulking was accomplished with either an extrapleural pneumonectomy (7 patients) or a lung-sparing pleurectomy-decortication (19 patients). Patients were injected with Foscan before surgery, and 652 nm light was delivered intraoperatively after completion of surgical debulking. Four light sensors were placed in the chest, allowing delivery of light to a uniform measured dose throughout the hemithorax. RESULTS: Four dose levels were explored. The MTD was 0.1 mg/kg of Foscan injected 6 days before surgery in combination with 10 J x cm(-2) 652 nm light. Dose limiting toxicity at the next higher dose was a systemic capillary leak syndrome leading to death in 2 of 3 patients treated at that dose. Other PDT-related toxicities included wound burns and skin photosensitivity. In all, 14 patients were treated at the MTD without significant complications. CONCLUSIONS: Foscan-mediated PDT can be safely combined with surgery at the established MTD. Unlike most other surgery-based multimodal treatments for mesothelioma, Foscan-mediated PDT affords the option, in selected patients, of accomplishing tumor debulking with a lung-sparing procedure rather than an extrapleural pneumonectomy. A phase II study is warranted. PMID- 12645725 TI - Risk factors for the development of postoperative complications after bronchial sleeve resection for malignancy: a univariate and multivariate analysis. AB - BACKGROUND: This study was designed to identify risk factors responsible for postoperative complications after bronchoplastic procedures. METHODS: Excluding sleeve pneumonectomies between January 1994 and December 2001, 108 patients underwent bronchoplastic procedures for bronchial malignancy. Prospectively documented data were age, gender, side, type of bronchial reconstruction, extended resection, histology, TNM stage, diseased lobe, and bronchial tumour occlusion. Cardiovascular (CV) risk factors included heart disease, arterial hypertension, cerebro-occlusive disease, peripheral artery disease of the lower extremities, diabetes mellitus, and abdominal aortic aneurysm. Patients were grouped according to the presence/absence of any CV risk factor and the absolute number of CV risk factors present (zero to four). Non-CV risk factors included neoadjuvant chemotherapy, alcoholism, lung disease, sleep apnea, history of recent pneumococcal sepsis, and repeat thoracotomy. Groups were assembled according to the presence or absence of any non-CV risk factor, neoadjuvant chemotherapy, and alcoholism. Respiratory risk factors included lung function and blood gas analysis. Groups were assembled according to the absolute number of respiratory risk factors in each person (zero to three) and the combination of respiratory and CV risk factors. Complications were defined as septic (pneumonia, empyema, brochopleural fistula, colitis) and aseptic. For univariate statistical analysis, t test, cross-tabulation, and chi2 test were used. All factors with a significance of p < 0.1 were entered into a binary backwards-stepwise logistic regression model. RESULTS: The combination of respiratory and CV risk factors (p = 0.012, OR = 0.165) was predictive for overall complications. Coronary artery disease (p = 0.02, OR = 0.062) and the combination of two respiratory risk factors (p = 0.008, OR = 0.062) were predictive for septic complications. Peripheral artery disease (p = 0.024, OR = 0.28), moderate (p = 0.01, OR = 0.13) and severe chronic obstructive pulmonary disease (p = 0.018, OR = 0.11), and extended resections (p = 0.003, OR = 0.017.) were predictive for aseptic complications. CONCLUSIONS: Comorbidity significantly influences the postoperative complication rate and is therefore crucial for evaluation of patients for bronchoplastic procedures. Different risk factors are responsible for the occurrence of septic and aseptic complications after bronchoplastic procedures. PMID- 12645724 TI - Videothoracoscopic bleb excision and pleural abrasion for the treatment of primary spontaneous pneumothorax: long-term results. AB - BACKGROUND: The goal of this study was to evaluate the long-term efficiency of videothoracoscopic bleb excision and pleural abrasion for the treatment of primary spontaneous pneumothorax. METHODS: From July 1991 to December 1997, 182 patients with primary spontaneous pneumothorax were treated by a single technique at our institution. Seven patients had single-stage bilateral procedures and 11 other patients had staged bilateral procedures. Indications for operation were first episode with prolonged air leak, incomplete lung reexpansion, or job restrictions (n = 59), first ipsilateral recurrence (n = 57), second or third ipsilateral recurrence (n = 34), contralateral recurrence (n = 25), synchronous bilateral pneumothorax (n = 3), hemopneumothorax (n = 3), and tension pneumothorax (n = 1). All patient data were reviewed retrospectively, and 167 patients were available for late follow-up (92%). RESULTS: Mean operative time was 57 +/- 19 minutes. Conversion to thoracotomy was required in 1 patient (0.6%). Mean duration of pleural drainage was 5.8 +/- 1.2 days (range, 4 to 26 days), and mean postoperative stay was 7.7 +/- 1.6 days (range, 6 to 31 days). Postoperative complications occurred in 50 patients (27.4%), the most frequent being prolonged air leak (14.8%), and in-hospital mortality was 0%. After a mean follow-up of 93 +/- 22 months (range, 57 to 134 months; median, 84 months), five ipsilateral recurrences were noted (3%). Three recurrences occurred within 12 months of videothoracoscopy and required reoperation. Two patients had partial pneumothorax recurrence at 39 and 58 months, and were treated conservatively with chest tube insertion and tale slurry. After 1 year, 10.7% of patients complained of chronic chest pain or discomfort, although none was taking pain medication after 3 months. Most patients (89.8%) were satisfied or very satisfied of their operation. All patients had returned to sport activities within 2 years. CONCLUSIONS: Videothoracoscopic bullectomy and pleural abrasion is a reliable and safe method to treat primary spontaneous pneumothorax. Long-term recurrences occur with an acceptable rate that compares with results after limited thoracotomy. Chronic chest pain or discomfort is unpredictable and may represent a problem in a few patients. PMID- 12645726 TI - Identifying patients at risk of early postoperative recurrence of lung cancer: a new use of the old CEA test. AB - BACKGROUND: In the current study, we report the carcinoembryonic antigen (CEA) capability to predict early tumor relapses after a pulmonary resection for nonsmall cell lung cancer (NSCLC). METHODS: We studied 118 consecutive NSCLC patients who were clinically judged operable and were eventually operated upon. Anthropometric, clinical, and CEA data along with the results of both preoperative and postoperative stage classifications were recorded. All patients were followed up for at least 1 year after surgery and the time to the first clinical recurrence recorded. Receiver-operating characteristic (ROC) curves and diagnostic formulas were used for data analysis. RESULTS: In this series the CEA test was among the most accurate methods to predict an early postoperative recurrence (ROC area: 0.72, 95% confidence interval [CI]: 0.60 to 0.85, p = 0.001; accuracy rate for CEA at the threshold of 10 ng/mL: 83%, CI: 76% to 90%). Also predictive was the postoperative pathologic stage of disease (ROC area: 0.68, CI: 0.56 to 0.80, p = 0.007). In tumors pathologically classified in stage Ia to IIb, a preoperative CEA level higher than 10 ng/mL was associated with a 67% probability of tumor relapse. In the same stages of disease, a CEA level less than 10 ng/mL increased the baseline probability of no recurrence from 80% to 88%. CONCLUSIONS: In operable patients with NSCLC the frequency of abnormal serum concentrations of CEA is low (17% in our series). However, it is important to identify such a small group of high-risk patients as many of them (in our study, 55% and 70% of those with a CEA value in excess of, respectively, 5 and 10 ng/mL) will develop an early postoperative recurrence. Such patients should be investigated preoperatively by mediastinoscopy or positron emission tomography in even in the absence of suspicious symptoms and signs. Then after an apparently successful operation, they should be carefully followed up. These patients could represent a suitable target for neoadjuvant clinical trials of selected high-risk groups. PMID- 12645727 TI - Complement activation and cytokine and chemokines release during mediastinitis. AB - BACKGROUND: Mediastinitis after open heart operation is an infrequent, but life threatening complication with a reported incidence rate between 1% and 4%. Hospital mortality is estimated at 10% to 35%. The aim of the present work was to study the systemic inflammatory reaction as judged by complement activation and cytokine and chemokines release in patients with mediastinitis after open heart operation. METHODS: Seven patients with clinical signs of mediastinitis were included. Three patients had undergone coronary artery bypass grafting, whereas 4 patients had combined coronary artery bypass grafting, valve replacement, or valvuloplasty. Blood samples were drawn before induction of anesthesia and at the time of reoperation, and thereafter daily during the hospital stay. Controls comprised similar patients with an uneventful postoperative course. RESULTS: The terminal SC5b-9 complement complex concentration in the mediastinitis patients was substantially higher compared with the controls (p < 0.001), and the terminal SC5b-9 complement complex values showed no overlap between the two groups. Interleukin-8, stromal cell-derived factor-1alpha and IL-6 concentrations were also significantly higher in the mediastinitis group than in the control group (p < 0.001), but with considerable overlap between the groups. Interleukin-1beta, interleukin-10, and monocyte chemoattractant protein-1 concentrations were slightly higher in the mediastinitis group, and no differences were seen for the tumor necrosis factor-alpha. CONCLUSIONS: During mediastinitis, the complement is activated and the cytokines and chemokines, interleukin-6, interleukin-8, and stromal cell-derived factor-1alpha are released. These proteins may be involved in the pathogenesis of this complication. Terminal SC5b-9 complement complex may be an indicator to discriminate mediastinitis patients from those with uneventful course. PMID- 12645729 TI - Perfadex for clinical lung procurement: is it an advance? AB - BACKGROUND: Extensive laboratory experience suggested that low potassium dextran lung preservation solution (Perfadex; Medisan, Uppsala, Sweden) is superior to Euro-Collins (EC; Frusen, Hamburg, Germany), the clinical standard. The purpose of this study was to evaluate Perfadex in clinical lung transplantation. METHODS: A retrospective analysis of the outcome of 69 consecutive lung allografts retrieved and used for transplantation was made. Donor lungs were flushed with EC in 37 patients and Perfadex in 32 patients. The evaluation measurements were quantitative chest roentgenogram score (grade 0 to 4), graft oxygenation, duration of mechanical ventilation, length of intensive care treatment, and survival. RESULTS: The mean chest roentgenogram score was 1.55 and 1.81 for the EC group compared with 1.18 and 2.09 for the Perfadex group at 1 and 48 hours, respectively (p = 0.1 and 0.8, respectively). Arterial alveolar oxygen tension ratio was similar at 12 and 24 hours (0.61 vs 0.67; p = 0.8; and 0.64 vs 0.53; p = 0.3, respectively). The mean ventilation time was 71.2 +/- 32.3 hours versus 81.9 +/- 43.6 hours for the EC and Perfadex groups, respectively (p = 0.4). The mean intensive therapy unit stay was 3.1 +/- 2.6 days for the EC group compared with 4.1 +/- 3.9 days for the Perfadex group (p = 0.4). Death caused by primary organ failure was 5.1% for the EC group compared with 3.1% for the Perfadex group (p = 0.8). CONCLUSIONS: There was no difference between Perfadex and EC in clinical lung preservation. This may reflect the difference between controlled laboratory environment and the real world of brain death lung injury. Further studies are required to investigate the impact of Perfadex in the long-term outcome of lung transplantation. PMID- 12645728 TI - Recurrent pleural effusion complicating liver cirrhosis. AB - BACKGROUND: Pleural effusion (PE) is a rare complication of advanced liver cirrhosis, which may lead to an operation when uncontrolled. The purpose of this study was to evaluate the modality of the occurrence of pleural effusion and to describe its surgical management. METHODS: We studied 21 patients who were referred to the department of thoracic surgery because of massive and recurrent PE caused by liver cirrhosis. The PE was a transudate in 16 patients and an exudate in 5. Talc pleurodesis was attempted in all patients. The patients were divided into two groups. Video assisted thoracoscopy was performed in 13 patients in whom the clinical condition permitted general anesthesia; the pleural cavity was entirely explored before pleurodesis (group 1). Chest tube drainage alone was performed in 8 patients who were unable to undergo general anesthesia; talc pleurodesis was performed through the chest tube in these patients (group 2). RESULTS: In group 1 the PE was right-sided in 8 patients, left-sided in 3, and bilateral in 2. Diaphragmatic defects were observed in 2 patients, and a fluid leak oozing from the diaphragm was observed in 1 patient. Ten patients were considered cured and were without recurrence. Two patients underwent late recurrence before dying from their liver cirrhosis. Only 1 patient had an early recurrence that was cured by complementary talc slurry. In group 2 all patients presented with a right PE; of these, 3 patients died from septic shock caused by pleural infection. Three patients underwent early recurrence but were cured after repeat talc slurry. One patient had a midterm recurrence. One patient had an early recurrence treated by intrahepatic porto-systemic shunt with partial improvement. CONCLUSIONS: Passage of ascites through diaphragmatic defects appears to be the main cause of PE complicating cirrhosis. Patients may benefit from talc pleurodesis. Video assisted thoracoscopy pleurodesis is the technique of choice with consistent results. Repeated talc injection through the drain may prove useful for patients in poor clinical status. PMID- 12645730 TI - Natural history of inadvertent aorta-saphenous vein-coronary vein bypass graft. AB - Inadvertent distal anastomosis of an aortocoronary bypass graft to a coronary vein is a rare but potentially serious complication of coronary artery bypass surgery. We describe a patient in whom such a conduit was discovered only incidentally 17 years after its creation. This case illustrates the pertinent features of this anomaly and demonstrates that it can have a benign natural history. PMID- 12645732 TI - Severe hemoptysis 6 years after coronary artery bypass grafting. AB - A patient with a history of coronary artery bypass grafting was admitted with severe hemoptysis. Bronchoscopy showed recent bleeding with clot formation in the lingular bronchus, but no tumor was visualized. Several biopsies of the underlying mucosa were negative. Coronary angiography showed patent venous and arterial bypass grafts. Selective angiography of the left internal mammary artery revealed one large and two smaller aberrant bronchial side branches, which probably caused the lingular hemorrhage. We performed embolization of the largest aberrant branch. After a follow-up of 3 months, hemoptysis had not recurred. PMID- 12645731 TI - Biventricular pacing with coronary bypass and Dor's ventriculoplasty. AB - We applied biventricular pacing to a patient with severe ischemic dilated cardiomyopathy (left ventricular [LV] ejection fraction 19%, LV end-diastolic volume 360 mL, and complete left bundle branch block). An epicardial LV lead was surgically implanted concomitant with on-pump beating coronary artery bypass grafting and Dor's endoventricular circular patch plasty. Biventricular pacing immediately achieved the resynchronization of the LV contraction, and improved cardiac function as well as reducing mitral regurgitation. Biventricular pacing combined with cardiac surgery for patients with cardiomyopathy and complete left bundle branch block may produce beneficial effects on LV function. PMID- 12645733 TI - A successfully treated case of blunt traumatic right coronary ostium rupture. AB - Cardiac tamponade due to coronary artery rupture, as a consequence of blunt trauma, is a rare but usually fatal condition. We successfully obtained primary hemostasis with emergency room thoracotomy, followed by delayed definitive treatment of the ruptured right coronary artery ostium in a motor vehicle accident victim with multifocal hemorrhagic lesions. Survival of patients with the described serious trauma has not been reported, and we discuss herein our treatment strategy. PMID- 12645734 TI - Aortic valve replacement in a patient with erythropoietic protoporphyria. AB - Erythropoietic protoporphyria (EPP) is a disorder of heme synthesis that causes excessive accumulation of protoporphyrin. The predominant clinical feature is photosensitivity triggered by light at wavelengths near 400 nm. We describe a 52 year-old man with EPP who underwent aortic valve replacement due to severe regurgitation. To prevent burn injuries, astral lamps in the operating room were covered with yellow film filters. Preoperative autologous blood donation was not undertaken. Blood priming of the extracorporeal circuit was performed to maintain adequate hemoglobin concentrations, which resulted in reduction of heme synthesis. The patient was discharged in good health without any signs or symptoms of EPP. PMID- 12645735 TI - Mechanical ventricular support lowers pulmonary vascular resistance in a patient with congential heart disease. AB - Severely elevated pulmonary vascular resistance (PVR) is a relative contraindication to orthotopic heart transplantation. A potential novel strategy to reverse elevated PVR may be implantation of a chronic left ventricular assist device with subsequent left ventricular unloading. We present a patient with elevated PVR secondary to congenital heart disease who was listed for heart-lung transplant. The patient underwent placement of biventricular assist devices and subsequently experienced marked reduction of PVR, ultimately enabling successful heart transplantation. PMID- 12645736 TI - Pulmonary artery banding before Norwood procedure. AB - Described here is the use of bilateral pulmonary artery banding as a means to achieve effective restoration of adequate systemic blood flow before a Norwood procedure in two newborns with hypoplastic left heart syndrome who presented after birth with a severe imbalance of Qp/Qs and multiorgan system dysfunction despite usual pharmacologic and ventilatory strategies. PMID- 12645737 TI - Pulmonary-to-systemic blood flow ratio oriented management after repair of obstructive total anomalous pulmonary venous connection in neonates with single ventricle. AB - Caval oxygen saturation was monitored to estimate pulmonary-to-systemic blood flow ratio after relief of obstructive total anomalous pulmonary venous connection in two neonates with single ventricle. Distribution between systemic and pulmonary blood flow was manipulated by pharmacologic, ventilatory, and surgical interventions aimed at achieving pulmonary-to-systemic blood flow ratio of 0.5 to 1.0. Monitoring of pulmonary-to-systemic blood flow ratio facilitates appropriate balancing between tissue perfusion and oxygenation, and detects redundant ventricular volume-load. PMID- 12645738 TI - Anatomic correction for corrected transposition after pulmonary unifocalization. AB - A Senning plus Rastelli operation was performed in a patient who had a rare combination of congenitally corrected transposition of the great arteries (S,L,L) with dextrocardia, major aortopulmonary collaterals, and diminutive central pulmonary arteries with arborization defects. The patient required four preparatory operations including bilateral unifocalizations of the aortopulmonary artery collaterals. Pulmonary artery to systemic pressure ratio after the double switch operation was 0.6. The patient demonstrates good biventricular function on echocardiogram at 3 months after the operation. PMID- 12645739 TI - Retroaortic innominate vein with coarctation of the aorta: surgical repair and embryology review. AB - A newborn girl with a retroaortic innominate vein, coarctation of the aorta, ventricular septal defect, and subaortic stenosis underwent a complete repair at 8 days of age. The ascending aorta was transected and the innominate vein was brought in front of it. We review the anatomy and embryology of this rare entity and describe the surgical technique to avoid recoarctation and innomante vein compression. PMID- 12645741 TI - Surgical resection of a solitary plasmacytoma originating in a rib of a patient with Castleman's disease. AB - Castleman's disease is a rare disorder characterized by lymphoid hyperplasia. It may present as asymptomatic involvement of one lymph node group or as a multicentric disease with systemic features. We report a patient with Castleman's disease who presented with axillary lymphadenopathy associated with a solitary plasmacytoma originating from a rib. The affected rib was surgically resected and radical radiotherapy was subsequently administered to the axillary lymph nodes. In this particular case, a joint surgical and oncologic approach resulted in a successful outcome. PMID- 12645740 TI - Development of brain metastasis 5 years before the appearance of the primary lung cancer: "messenger metachronous metastasis". AB - We report a patient with a brain metastasis that presented 5 years before the primary adenocarcinoma of the lung from which it originated. The metastasis and the primary tumor were removed. To confirm their common origin, we used comparative genomic hybridization. We have named this type of metastasis "messenger metachronous metastasis." The patient remains well 79 months after the brain metastasectomy and 18 months after the lung surgery. PMID- 12645742 TI - Mucoepidermoid carcinoma in unilateral hypoplastic lung: a rare tumor in a rarer condition. AB - We present a rare case of a mucoepidermoid carcinoma arising from a unilateral isolated hypoplastic lung treated with a left pneumonectomy. The rarity of the condition is described and both pathologies discussed. PMID- 12645744 TI - Postpneumonectomy syndrome in single lung transplantation recipient following previous pneumonectomy. AB - Successful single lung transplantation following previous pneumonectomy has not been reported in the literature. We report a patient with cystic fibrosis who underwent left single lung transplantation following right pneumonectomy 13 years previously. The outcome was adversely affected by postpneumonectomy syndrome. PMID- 12645743 TI - Sclerosing hemangioma isolated to the mediastinum. AB - Sclerosing hemangioma is an uncommon tumor of unknown histogenesis that generally develops in the lung. We report on a 48-year-old woman with a sclerosing hemangioma that was apparently isolated to the mediastinum. To our knowledge, sclerosing hemangioma arising in the mediastinum has not been previously reported. Potential mechanisms explaining the isolation of sclerosing hemangioma in the mediastinum are discussed. PMID- 12645745 TI - Rare but dangerous anomaly of the right pulmonary vein in subcarinal dissection. PMID- 12645747 TI - Implantation of the Jarvik 2000 left ventricular assist device without the use of cardiopulmonary bypass. AB - In this report, we describe successful implantation of a Jarvik 2000 left ventricular assist device (Jarvik Heart, Inc, New York, NY) without the use of cardiopulmonary bypass in a patient who was a member of the Jehovah's Witness faith. To accomplish this, we had to change our implantation technique. The modified technique, which minimizes the risk of bleeding and end-organ dysfunction, can also be used to decrease cardiopulmonary bypass time. PMID- 12645746 TI - Percutaneous unknotting and retrieval of Swan-Ganz catheter. PMID- 12645749 TI - Aortic arch reconstruction using a trifurcated graft. AB - Cerebral protection and prevention of atheroembolism remain challenges in aortic arch reconstruction. To reduce neurologic complications, we developed a "no touch" technique in which a trifurcated graft is anastomosed to the arch vessels during hypothermic circulatory arrest, reducing the risk of embolization while minimizing cerebral ischemia by permitting antegrade cerebral perfusion as arch reconstruction is completed. PMID- 12645748 TI - Valve-sparing root reduction plasty in aortic aneurysm: the "Jena" technique. AB - Reconstruction of the aortic valve and replacement of the ascending aorta instead of use of a composite graft is the preferred method in patients, who demonstrate an ascending aneurysm accompanied by aortic root enlargement and aortic valve insufficiency with morphologically intact cusps. Two techniques inaugurated by David and Yacoub have gained widespread acceptance. However, both approaches are technically demanding. We present a simplified alternative technique for aortic valve reconstruction by means of a reduction plasty of the aortic root without the necessity of transection maneuvers. PMID- 12645750 TI - A practical tool to control bleeding during sternal reentry for pseudoaneurysm of the ascending aorta. AB - Big pseudoaneurysms of the ascending aorta after a previous thoracic operation are rare and represent a surgical challenge. Because the rupture during sternal reentry occurs before the clamp-control of the distal ascending aorta, it is essential to control the bleeding until the adhesions are released in order to place the sternal retractor. We report the use of Foley catheter with a malleable guidewire to control the bleeding from the defect in the ascending aorta causing a pseudoaneurysm in case of a limited access. PMID- 12645751 TI - Development of tracheal surgery: a historical review. Part 2: Treatment of tracheal diseases. PMID- 12645752 TI - The volume-outcome relationship: from Luft to Leapfrog. AB - Numerous reports have documented a volume-outcome relationship for complex medical and surgical care, although many such studies are compromised by the use of discharge abstract data, inadequate risk adjustment, and problematic statistical methodology. Because of the volume-outcome association, and because valid outcome measurements are unavailable for many procedures, volume-based referral strategies have been advocated as an alternative approach to health-care quality improvement. This is most appropriate for procedures with the greatest outcome variability between low-volume and high-volume providers, such as esophagectomy and pancreatectomy, and for particularly high-risk subgroups of patients. Whenever possible, risk-adjusted outcome data should supplement or supplant volume standards, and continuous quality improvement programs should seek to emulate the processes of high-volume, high-quality providers. The Leapfrog Group has established a minimum volume requirement of 500 procedures for coronary artery bypass grafting. In view of the questionable basis for this recommendation, we suggest that it be reevaluated. PMID- 12645753 TI - Congenital diaphragmatic hernia. PMID- 12645754 TI - Surgical glue and necrosis of arterial wall. PMID- 12645755 TI - What are the risks of using biologic glues? PMID- 12645756 TI - Alternate technique of routing the in situ right internal mammary artery to graft the left anterior descending artery and its branches. PMID- 12645757 TI - Which is the best sternotomy closure technique? PMID- 12645758 TI - The use of Z scores in assessing neuropsychological change after cardiac operations. PMID- 12645759 TI - 25-year follow-up of homograft aortic conduits for coarctation repair. PMID- 12645761 TI - Late recurrence after resection of stage I lung adenocarcinoma. PMID- 12645762 TI - A favorable way to close the bronchus in pneumonectomy. PMID- 12645760 TI - Preoperative preparation of patients with myasthenia gravis forestalls postoperative respiratory complications after thymectomy. PMID- 12645763 TI - Diffusive sampling of radon. AB - Information required for the design and testing of diffusive samplers for radon is scattered across numerous disciplines, making it difficult to use. To aid future research, a multidisciplinary review is given addressing the following: 1) the differential equations describing mass transfer in a radon diffusive sampler; 2) mathematical procedures for solving these equations; 3) the physical parameters defining the adsorbent in the diffusive sampler; and 4) a standardized protocol for validating the performance of diffusive samplers for radon. PMID- 12645764 TI - Results of a study to determine the impact of the new dosimetry performance standard, N13.11-2001. AB - Dosimeters were irradiated using the protocol described in the new ANSI HPS N13.11-2001 standard, "American National Standard for Dosimetry--Personnel Dosimetry Performance--Criteria for Testing." In particular, the following hypotheses were evaluated: (1) expanding the scope of testing fields in the x-ray category would not adversely impact the performance of dosimeter systems; (2) incorporating routine angular testing for medium-energy and high-energy photon fields would not adversely impact the performance of dosimeter systems; (3) implementing test criteria would adversely impact the performance of dosimeter systems; and (4) the narrow spectrum x-ray fields specified in the standard would provide adequate replacements for analogous k-fluorescence techniques previously used by DOELAP. The study tested only subcategories IIA, General photon fields, and IID, Plutonium-specific fields, and did not address mixed irradiations. The results of the study supported hypotheses (1) and (2) above. The results of the study indicated that the individual dosimeter performance test (10% rule) did not seriously impact the performance of dosimeter systems nor did results support the use of NS20 as a replacement x-ray field for the 17 keV k-fluorescence x-ray field. PMID- 12645766 TI - Calculated organ doses for Mayak production association central hall using ICRP and MCNP. AB - As part of an ongoing dose reconstruction project, equivalent organ dose rates from photons and neutrons were estimated using the energy spectra measured in the central hall above the graphite reactor core located in the Russian Mayak Production Association facility. Reconstruction of the work environment was necessary due to the lack of personal dosimeter data for neutrons in the time period prior to 1987. A typical worker scenario for the central hall was developed for the Monte Carlo Neutron Photon-4B (MCNP) code. The resultant equivalent dose rates for neutrons and photons were compared with the equivalent dose rates derived from calculations using the conversion coefficients in the International Commission on Radiological Protection Publications 51 and 74 in order to validate the model scenario for this Russian facility. The MCNP results were in good agreement with the results of the ICRP publications indicating the modeling scenario was consistent with actual work conditions given the spectra provided. The MCNP code will allow for additional orientations to accurately reflect source locations. PMID- 12645765 TI - Absorbed dose to very low birth weight infants from 18F-fluorodeoxyglucose. AB - The radiation absorbed dose to very low birth weight infants from 18F fluorodeoxyglucose was investigated. Ten newborns undergoing clinical positron tomography lung imaging were included in this study. Two consecutive 45-min dynamic scans immediately following intravenous injection of fluorodeoxyglucose were acquired; the first was over the head, and the second was over the chest. Time-activity curves were generated for the brain, heart wall, lungs, and, when visible, the kidneys. The cumulated activity measurements obtained were for the entire organ masses; these masses were much smaller than the corresponding organ masses for the newborn mathematical model. Patient-specific dosimetry yielded average doses of 2.5 x 10(-1) mGy MBq(-1) for the brain, 6.8 x 10(-1) mGy MBq(-1) for the heart wall, 2.2 x 10(-1) mGy MBq(-1) for the kidneys, and 4.4 x 10(-1) mGy MBq(-1) for the lungs. The effective dose was estimated to be 2.1 x 10(-1) mSv MBq(-1), which is half that previously published for newborns but still an order of magnitude higher than that for adults. PMID- 12645767 TI - Inconstant sun: how solar evolution has affected cosmic and ultraviolet radiation exposure over the history of life on Earth. AB - Four billion years ago, sea-level UV exposure was more than 400 times as intense as today, the dose from solar cosmic rays was five times present levels, and galactic cosmic rays accounted for only about 10% their current contribution to sea-level radiation doses. Exposure to cosmic radiation accounts for about 10% of natural background radiation exposure today and includes dose from galactic cosmic rays and solar charged particles. There is little exposure to ionizing wavelengths of UV due to absorption by ozone. The sun has evolved significantly over its life; in the past there were higher levels of particulate radiation and lower UV emissions from the sun, and a stronger solar wind reduced radiation dose in the inner solar system from galactic cosmic rays. Finally, since the early atmosphere contained little to no oxygen, surface levels of UV radiation were far higher in the past. PMID- 12645768 TI - Age-specific uncertainty of the 131I ingestion dose conversion factor. AB - The production of weapons-grade nuclear materials and their by-products has resulted in a number of releases from United States Department of Energy facilities. 131I, a fission by-product, is one of the most common radionuclides generated and released to the environment. It is known that there are differences in various physiological parameters over all age groups when considering biokinetic modeling of iodine. The establishment of age-specific dose conversion factor uncertainty is necessary for accurate internal dose assessment. The 131I dose conversion factor determined herein is log-normally distributed with varying age-specific distribution characteristics. The two most important parameters for determination of the dose conversion factor, in all age groups, are thyroid mass and iodine uptake fraction. These parameters are assumed to be highly correlated with a relationship that is quite important to dose conversion factor uncertainty. Dose estimates to individuals exposed to radioiodine can be determined more accurately with an increased understanding of the correlation between thyroid mass and uptake fraction. Improved dose estimates following oral intakes of 131I can be made from the consideration of age-specific dose conversion factors and their input parameters. PMID- 12645769 TI - Vertical profile of tritium concentration in air during a chronic atmospheric HT release. AB - The vertical profiles of tritium gas and tritiated water concentrations in air, which would have an influence on the assessment of tritium doses as well as on the environmental monitoring of tritium, were measured in a chronic tritium gas release experiment performed in Canada in 1994. While both of the profiles were rather uniform during the day because of atmospheric mixing, large gradients of the profiles were observed at night. The gradient coefficients of the profiles were derived from the measurements. Correlations were analyzed between the gradient coefficients and meteorological conditions: solar radiation, wind speed, and turbulent diffusivity. It was found that the solar radiation was highly correlated with the gradient coefficients of tritium gas and tritiated water profiles and that the wind speed and turbulent diffusivity showed weaker correlations with those of tritiated water profiles. A one-dimensional tritium transport model was developed to analyze the vertical diffusion of tritiated water re-emitted from the ground into the atmosphere. The model consists of processes of tritium gas deposition to soil including oxidation into tritiated water, reemission of tritiated water, dilution of tritiated water in soil by rain, and vertical diffusion of tritiated water in the atmosphere. The model accurately represents the accumulation of tritiated water in soil water and the time variations and vertical profiles of tritiated water concentrations in air. PMID- 12645770 TI - Nationwide survey of radon levels in Korea. AB - A nationwide radon survey was conducted to provide data on the annual average indoor radon concentration in Korean homes. This survey also provided data on the variation of radon concentration with season, house type, and building age. The arithmetic mean (AM) of annual radon concentration in Korean homes was 53.4 +/- 57.5 Bq m(-3). The indoor radon concentration showed a lognormal distribution with a geometric mean (GM) and its standard deviation (GSD) of 43.3 +/- 1.8 Bq m( 3). The radon concentrations in the traditional Korean-style houses were about two times higher than those in apartments and row houses. The average annual outdoor radon concentration was 23.3 Bq m(-3). The average annual effective dose to the general public from radon was 1.63 mSv y(-1). PMID- 12645771 TI - Tritium release from nuclear power plants in Taiwan. AB - Tritium is produced in light water reactors from various sources and the design source terms for both BWR and PWR plants are reviewed. The chemical forms of tritium produced in the coolant are discussed in terms of water radiolysis and free radical reactions in the reactor core regions, and the major form of tritium released from the coolant systems is confirmed to be tritiated water. The tritium activity concentrations and inventories in various coolant systems have been measured, and the release pathways of tritium from nuclear power plants are also reviewed in this paper. Decreasing trends of tritium release from nuclear power plants in both liquid waste and ventilation sources have been observed in Taiwan. The impact of tritium release on environmental radiation is estimated with well established screening models, and the results confirm that the impact is less than 1% of the regulatory limits and less than 0.1% of the natural radiation background. PMID- 12645772 TI - Natural soil gamma radioactivity levels and resultant population dose in the cities of Zacatecas and Guadalupe, Zacatecas, Mexico. AB - The study of natural gamma radioactivity was made to determine the concentrations of natural radionuclides in soil. Twenty soil samples collected in the cities of Zacatecas and Guadalupe and their suburban areas in the Mexican state of Zacatecas were analyzed by gamma-ray spectrometry to determine the activity concentrations of 226Ra, 232Th, and 40K. Gamma-spectrometry measurements were made using a hyperpure germanium detector surrounded with shielding material to reduce the background counting rate. The GammaVision-32 MCA emulation software was used for gamma-ray spectrum analysis and the TRUMP card of 2k as a MCA emulator. Conversion factors were used to calculate the dose to the population from outdoor exposure to terrestrial gamma rays. The measured activity concentration of 226Ra varies from 11 to 38 Bq kg(-1), the activity concentration of 232Th varies from 8 to 38 Bq kg(-1). The activity concentration of 40K is in the range 309-1,049 Bq kg(-1). The overall population mean outdoor terrestrial gamma dose rate is 44.94 nGy h(-1). PMID- 12645773 TI - Time resolved photoluminescence from a phosphate glass (GD-300) irradiated with heavy ions and gamma rays. AB - The time-resolved photoluminescence from a phosphate-glass photoluminescence dosimeter (GD-300) was compared for different quality radiations: relativistic heavy ions (12C and 40Ar) and 60Co gamma rays. The intensities of photoluminescence afterglow at 2-7 micro(s) after pulsed UV excitation, i.e., in the time range used for conventional dosimetry, soon reached stable conditions for all radiations. Whereas the early photoluminescence emission at the range less than 0.4 micro(s) was notably unstable for gamma rays only; it continued decreasing even at 2 d after irradiation. In contrast, the photoluminescence afterglows for the heavy ions were stable over the whole range after excitation. These results indicate that the process of photoluminescence-center formation is different for heavy ions and gamma rays. PMID- 12645775 TI - Safety and security of radiation sources in the aftermath of 11 September 2001. PMID- 12645774 TI - Characterization of the angular response of an "isotropic" MOSFET dosimeter. AB - MOSFET dosimeters are seeing increased use for various dosimetry applications. Previously available commercial MOSFET dosimeters demonstrate significant anisotropies in the response when irradiated from certain directions at diagnostic x-ray energies. The angular response of a more recently introduced MOSFET dosimeter that claims an isotropic response to incident radiation is characterized. Measurements of the Thomson-Nielsen model TN-502RDI dosimeter were made for rotations of 360 degrees in 15 degrees increments about both the axial and normal-to-axial axes. These measurements demonstrated that the model TN 502RDI dosimeter has a nearly isotropic response at diagnostic x-ray energies with variations of less than 3% deviation from the mean response for radiation incident from most directions. Only two specific orientations showed significant deviation from the overall isotropic response. These correspond to the exposure scenarios where radiation is incident directly along the axis where the wire leads attach to the MOSFET device and the distal tip, the 90 degrees and 270 degrees orientations for normal-to-axial rotations, respectively. The largest deviations from an isotropic response occur when the dosimeter is irradiated free in-air. Irradiations performed at the center of a tissue equivalent cylinder resulted in smaller deviations in angular response. The improved angular response of the TN-502RDI allows greater freedom in placement and use of MOSFET dosimeters in diagnostic radiology applications. PMID- 12645778 TI - The value of information technology in healthcare. AB - Not only will healthcare investments in information technology (IT) continue, they are sure to increase. Just as other industries learned over time how to extract more value from IT investments, so too will the healthcare industry, and for the same reason: because they must. This article explores the types of business value IT has generated in other industries, what value it can generate in healthcare, and some of the barriers encountered in achieving that value. The article ends with management principles for IT investment. PMID- 12645776 TI - Guidance on determining compliance of exposure to pulsed and complex non sinusoidal waveforms below 100 kHz with ICNIRP guidelines. PMID- 12645779 TI - ROI under scrutiny: the radical redefinition of a core concept. AB - This article offers a three-part analysis for identifying and assessing return on investment in healthcare information technology (IT) projects. Returns to IT can be financial, clinical, or structural. The goal is to identify key areas of measurable returns to both assess the value of a project before it is undertaken and to assess the actual value returned to the organization. Given the choice, many senior executives still prefer to rely on classic financial analyses, but the true value of a project is often found in the clinical and structural returns. Still, project-specific assessments do not answer the core question of whether long-term strategic investment in IT leads to a systemic strategic advantage to healthcare organizations. This article addresses these issues and indicates opportunities for further investigation. PMID- 12645780 TI - Can IT transform healthcare? PMID- 12645781 TI - Building the information bridge to cross the quality chasm. PMID- 12645782 TI - The importance of revisiting ROI. PMID- 12645783 TI - Acid rereflux: a review, emphasizing detection by impedance, manometry, and scintigraphy, and the impact on acid clearing pathophysiology as well as interpreting the pH record. AB - Acid clearing, the interval while intraesophageal pH is < 4 after a traditional acid reflux event (RE), is a potential "blind spot" during pH monitoring, when reflux of acidified gastric contents may occur undetected by the pH probe. This is termed "acid rereflux." Acid rereflux comprised 61% (169/262) of acid REs in recumbent postprandial patients with severe GERD in two reports using simultaneous pH monitoring and manometry as well as multichannel intraluminal impedance (MII) in one, and scintigraphy in the other. Acid rereflux events often recurred with short intervals between them. The pH probe alone was insufficient to detect most acid rereflux REs, since expanding pH criteria for an acid RE (> 1 unit fall while pH < 4) detected only 35% of acid rereflux REs. When a variety of patients and study conditions was examined, simultaneous manometry-pH monitoring found more frequent acid rereflux in the following situations: (1) patients with vs those without esophagitis; (2) recumbent vs upright posture, and (3) postprandial vs preprandial. Of pathophysiologic importance, acid rereflux in the blind spot is the most common cause of prolonged daytime acid REs in GERD patients. Of clinical importance, the 24-hr pH parameter "% acid exposure" should be relied upon most in interpreting the 24-hr pH record, because those parameters that relate to RE frequency may be inaccurate due to acid rereflux REs that are not counted. Furthermore, identifying as many REs as possible gives a more reliable indication of the severity of antireflux barrier incompetence, as well as more REs to correlate with patients symptoms that should improve sensitivity of the symptom index. Ambulatory simultaneous pH monitoring and MII will allow these and other roles for acid rereflux to be assessed during the patients normal day. PMID- 12645785 TI - Validation of a new method of measuring esophageal acid exposure: comparison with 24-hour pH monitoring. AB - Recently, we developed a disposable acid exposure sensor whose in vitro response to acid below pH 4 is linearly determined by the duration of exposure and the degree of acidity. The aim of the present study was to compare the SR to simultaneous esophageal pH and duodenogastroesophageal reflux (DGER) monitoring (Bilitec) in patients investigated for presumed gastroesophageal reflux disease (GERD). Twenty-six patients (16 men, mean age 46 +/- 2 years) with symptoms suggestive of GERD underwent 24-hr ambulatory pH monitoring and SR monitoring at 5 cm proximal to the LES. DGER monitoring was performed in 21 patients. Exposure of the esophagus to acid and to DGER were analyzed. These data were compared to SR. A significant correlation was found between the exposure of the distal esophagus to acid and SR (R = 0.85; P < 0.0001). Similarly, the area below a cutoff pH 4 was significantly correlated to SR (r = 0.81; P < 0.0001). SR was not correlated to DGER (r = 0.16; NS). At a cutoff of 50, the sensitivity and specificity of SR to predict esophageal acid exposure > 5% of time were 91% and 93%, respectively conclusion, the response of the acid exposure sensor is strongly correlated with the results of simultaneous esophageal pH monitoring. The sensor seems able to reliably predict pathological esophageal acid exposure. These findings warrant larger studies of the clinical potential of the acid exposure sensor in the diagnosis and quantification of GERD. PMID- 12645784 TI - Streamlining 24-hour pH study for GERD: Use of a 3-hour postprandial test. AB - At present, the ambulatory 24-hr pH test has been used as a diagnostic tool to assess gastroesophageal reflux disease (GERD) in those patients with reflux symptoms and a normal endoscopy. However, patients poorly tolerate the prolonged nature of the 24-hr test. The aim of this study was to determine whether analyzing a 3-hr postprandial period from a full 24-hr study would be as sensitive as the longer test. Data were analyzed from a standard ambulatory 24-hr pH recording. A positive test was determined if the pH was < 4 for more than 4% of the study period with the probe placed 5 cm above the lower esophageal sphincter for both groups. The data were then reanalyzed by determining the percent time of pH < 4 during a 3-hr postprandial period. The results of 50 patients with a positive 24-hr test were compared with 50 patients with normal tests. The meal that was used to study the 3-hr postprandial period occurred in the late afternoon or early evening. The 3-hr postprandial test had a sensitivity of 88% when compared to the 24-hour test and a specificity of 98%. The positive predictive value was 100% for the 3-hr test, and the accuracy of this shorter test when compared with the standard 24-hour test was 95%. In conclusion the 3-hr postprandial analysis is a highly sensitive and specific test for demonstrating GERD. By using the shorter test, patient discomfort may be reduced and compliance enhanced. PMID- 12645786 TI - Eosinophilic esophagitis: strictures, impactions, dysphagia. AB - Eosinophilic esophagitis, long known to be a feature of acid reflux, has recently been described in patients with food allergies and macroscopically furrowed esophagus. The pathophysiology and optimal management of patients with eosinophilic esophagitis is unclear. We describe our clinical experience related to eosinophilic esophagitis and obstructive symptoms in children and propose etiopathogenesis and management guidelines. Twelve children with obstructive esophageal symptoms (11 male), median age 5 years, and identified to have eosinophilic esophagitis with > 5 eosinophils per high-power field (eos/hpf) are reported. Of these, four had strictures, six had impactions, and two had only dysphagia. A diagnostic evaluation included esophagogastroduodenoscopy with biopsies in all and upper gastrointestinal series, IgE, radioallergosorbent tests, and skin tests for food allergies in some cases. Esophageal histology specimens were independently analyzed for eosinophil density by two authors. Four of five children with > 20 eos/hpf responded to elimination diets/steroids. The fifth child responded to a fundoplication. Seven children had 5-20 eos/hpf and three of them with no known food allergies responded to antireflux therapy alone. Three others in this group with positive food allergies responded to treatment with elimination diets and/or steroids. The seventh patient in this group was lost to follow-up. In conclusion, on the basis of response to therapy, eosinophilic esophagitis can be subdivided into two groups: those with likely gastroesophageal reflux disease if < 20 eos/hpf and no food allergies, and others with allergic eosinophilic esophagitis associated with food allergies and often with > 20 eos/hpf. PMID- 12645787 TI - Isolated neonatal swallowing dysfunction: a case series and review of the literature. AB - Our purpose was to describe the natural history of isolated neonatal swallowing dysfunction (INSD). Nine infants with INSD are described. Eight presented within two weeks of birth. Symptoms included choking and cyanotic spells with feeds, recurrent aspiration, apnea, stridor, and vomiting. Three had nonspecific neurological abnormalities and were diagnosed later in life with underlying disorders (myotonic dystrophy, CHARGE association, velocardiofacial syndrome). All required tube feeding. Six tolerated nasogastric feedings and received a gastrostomy tube. Three failed nasogastric feeds and required jejunal feedings. The gastrostomy tube was removed in 7/9 at a mean age of 37 +/- 9 months. In conclusion, INSD has a good long-term prognosis. The presence of minor neurological abnormalities at presentation suggests another underlying disorder. Nasogastric feeding followed by a gastrostomy is recommended in those without gastroesophageal reflux. Jejunal feedings are necessary in some. While most improve over time, they may need nutritional support for 3 years or more. PMID- 12645789 TI - Relationship between gastric disease and deletion of cag pathogenicity island genes of Helicobacter pylori in gastric juice. AB - The cag pathogenicity island genes of Helicobacter pylori (ie, cag1, cag5, cagT, cagE, and cagA) were detected by PCR in DNA extracted from endoscopically collected gastric juice, and the relationship between these genes and gastric disease was studied in 25 patients with early gastric cancer, 9 patients with gastric ulcer, and 15 patients with chronic active gastritis. In three patients with early gastric cancer and one patient with gastric ulcer, cag pathogenicity island genes were amplified although H. pylori was not detected by conventional methods. Compared with conventional methods, the sensitivity of detection of cag genes was 92.3% (36/39) and the specificity was 60% (6/10). Among the patients with cagA amplification, only cagE was not amplified in one case each with early cancer and chronic active gastritis. In addition, none of cag1, cag5, cagT, and cagE were amplified in spite of cagA amplification in one patient with gastric ulcer. This method is a simple procedure, has a high sensitivity, and appears to be useful for accurate assessment of infection with cagA-positive strains. Because deletion of cag PAI genes was found in the patients with all three gastric diseases that we studied, it was suggested that the pathogenicity of H. pylori might not be determined by cag PAI genes in those cases. PMID- 12645788 TI - Gene expression of ornithine decarboxylase, cyclooxygenase-2, and gastrin in atrophic gastric mucosa infected with Helicobacter pylori before and after eradication therapy. AB - H. pylori (Hp) -induced atrophic gastritis is a well-known risk factor for the development of gastric cancer. Whether Hp eradication can prevent or retard the progress of atrophy and metaplasia has been the topic of numerous studies but the subject remains controversial. Recently, the increased expression of ornithine decarboxylase (ODC), gastrin and cyclooxygenase (COX)-2 has been shown to be increased in premalignant lesions in gastric mucosa and to play an essential role in the malignant transformation. The aim of the study is to assess the effect of eradication therapy on atrophic gastritis and analyze the gene expression for ODC, COX-2 and gastrin in gastric mucosa after succesful eradication in patients with atrophic gastritis. Twenty patients with chronic atrophic gastritis including both corpus and antrum of the stomach were included in this study. Four antral mucosal biopsy specimens were obtained from antrum and four from corpus. The histopathologic evaluation of gastritis was based on Sydney classification of gastritis. All patients were Hp positive based on the [13C] urea breath test (UBT) and the presence of anti-Hp IgG and anti-CagA-antibodies detected by ELISA. The patients were then eradicated with triple therapy consiting of omeprazol (2 x 20 mg), amoxycillin (2 x 1 g) and clarithromycin (2 x 500 mg) for seven days and vitamin C 1 g/day for three months. In gastric mucosal samples obtained from the antrum and corpus before and after eradication, the mRNA expression for ODC, COX 2, and gastrin was assessed by reverse-transcription polymerase chain reaction (RT-PCR). In all patients the gastric secretory analysis was performed by measuring gastric acid output and serum gastrin levels. After triple therapy the successful eradication assessed by UBT was observed in 95% of patients. In 45% of patients the infection with CagA-positive Hp strain was observed. Three months after eradication a significant reduction in the gastric activity (neutrophilic infiltrate) and severity (mononuclear infiltrate) of gastritis was observed. The atrophy score improved in both antrum and corpus after eradication. The expression of COX-2 and ODC was significantly up-regulated in the gastric mucosa of patients with atrophic gastritis and significantly reduced after eradication therapy. In all successfully eradicated patients with atrophic gastritis a significant increase in gastric acid secretion and decrease in serum gastrin were observed. We conclude that: (1) Hp eradication leads to the decrease in ODC and COX-2 gene expression in the gastric mucosa, and this may be relevant for the prevention of the Hp-associated gastric carcinogenesis; and (2) gastric atrophy ameliorates upon successful Hp eradication therapy. PMID- 12645791 TI - Neural mechanism of acupuncture-induced gastric relaxations in rats. AB - Acupuncture has been used to treat gastrointestinal symptoms in China for more than 3000 years. However, the mechanism of the beneficial effects of acupuncture remains unknown. Strain gauge transducers were implanted on the serosal surface of the stomach to record circular muscle contractions in thiobutabarbital anesthetized rats. Acupuncture on the right lower abdomen caused a transient relaxation of the stomach. Acupuncture-induced gastric relaxations were abolished by guanethidine, propranolol, splanchnic ganglionectomy, spinal cord transection, and spinomedullary transection. In contrast, N(G)-nitro-L-arginine, phentolamine, truncal vagotomy, and pontomedullary transection had no effect. Acupuncture increased the number of c-Fos immunopositive cells at the ventrolateral medulla (VLM). It is concluded that acupuncture-induced gastric relaxations are mediated via the somatosympathetic reflex. Its afferent limb is composed of abdominal cutaneous and muscle afferent nerves. Its efferent limb is the gastric sympathetic nerve and the reflex center is within the medulla. VLM neurons may play an important role in mediating this reflex. PMID- 12645790 TI - A novel vitamin E derivative (TMG) protects against gastric mucosal damage induced by ischemia and reperfusion in rats. AB - The aim of the present study was to investigate the antioxidative effects of water-soluble vitamin E derivative, 2-(alpha-D-glucopyranosyl)methyl-2,5,7,8 tetramethylchroman-6-ol (TMG), on ischemia-reperfusion (I/R) -induced gastric mucosal injury in rats. Gastric ischemia was induced by applying a small clamp to the celiac artery and reoxygenation was produced by removal of the clamp. The area of gastric mucosal erosion, the concentration of thiobarbituric acid reactive substances, and the myeloperoxidase activity in gastric mucosa significantly increased in I/R groups compared with those of sham-operated groups. These increases were significantly inhibited by pretreatment with TMG. The contents of both mucosal TNF-alpha and CINC-2beta in I/R groups were also increased compared with the levels of those in sham-operated groups. These increases of the inflammatory cytokines were significantly inhibited by the treatment with TMG. It is concluded that TMG inhibited lipid peroxidation and reduced development of the gastric mucosal inflammation induced by I/R in rats. PMID- 12645793 TI - Abnormalities of left colonic motility in ambulant nonconstipated patients with irritable bowel syndrome. AB - Our objective was to evaluate left colonic motility patterns recorded under physiological conditions during 24 hr in fully ambulant nonconstipated IBS patients compared to healthy controls. A 42-hr manometry of the left colon was performed in 11 nonconstipated IBS patients and 10 age- and sex-matched healthy volunteers. On day 1, a 6-channel, 10-cm interval, solid-state catheter was positioned. Frequency, amplitude, and motility index (MI) of segmenting pressure waves in the descending and sigmoid colon were calculated during the 24-hr study period on day 2. High-amplitude propagated contractions (HAPCs) were identified visually and their characteristics were calculated. In IBS patients a higher frequency of segmenting pressure waves was observed in the sigmoid colon compared to the descending colon (P = 0.006). In contrast, no regional differences were observed in controls. Awakening (P = 0.048) as well as having a meal (P = 0.024) was associated with a smaller increase of contraction frequency in the descending colon of IBS patients compared to controls. HAPCs occurred more frequently in IBS patients than in controls (P = 0.035). HAPCs in IBS patients reached a more distal colonic level and occurred more frequently in clusters. Defecation in IBS patients, but not in controls was always preceded by a cluster of HAPCs. In conclusion, left colonic segmenting pressure waves and HAPC characteristics are altered in nonconstipated IBS patients. PMID- 12645792 TI - Effects of rabeprazole sodium on gastric emptying, electrogastrography, and fullness. AB - Proton pump inhibitors have been reported to delay gastric emptying, but this effect is controversial. Our aim was to determine the effect of rabeprazole sodium on several parameters of gastric function including gastric emptying, myoelectrical activity and ingested water volume required to produce fullness. Fifteen healthy males underwent assessment of solid-phase gastric emptying with the [13C] Spirulina platensis breath test as well as electrogastrography and satiety testing using a 5-min water load. Subjects were evaluated at baseline, after administration of placebo, and after rabeprazole sodium 20 mg daily for one week. No significant differences were seen between groups with respect to solid phase gastric emptying as measured by T1/2 or T(lag). No differences were seen between baseline, placebo, and rabeprazole with respect to the number of normal electrogastrograms and the volume of water required to produce fullness. In conclusion, one week of therapy with rabeprazole sodium does not significantly alter gastric emptying, myoelectrical activity or threshold to fullness. PMID- 12645794 TI - Intestinal malabsorption as a manifestation of obstructive sleep apnea. PMID- 12645795 TI - Methane production during lactulose breath test is associated with gastrointestinal disease presentation. AB - It has recently been determined that there is an increased prevalence of bacterial overgrowth in IBS. Since there are two gases (hydrogen and methane) measured on lactulose breath testing, we evaluated whether the different gas patterns on lactulose breath testing coincide with diarrhea and constipation symptoms in IBS and IBD. Consecutive patients referred to the gastrointestinal motility program at Cedars-Sinai Medical Center for lactulose breath testing were given a questionnaire to evaluate their gastrointestinal symptoms. Symptoms were graded on a scale of 0-5. Upon completion of the breath test, the results were divided into normal, hydrogen only, hydrogen and methane, and methane only positive breath tests. A comparison of all subjects and IBS subjects was undertaken to evaluate diarrhea and constipation with regards to the presence or absence of methane. This was further contrasted to Crohn's and ulcerative colitis (UC) patients in the database. After exclusion criteria, 551 subjects from the database were available for comparison. Of the 551 subjects (P < 0.05, one-way ANOVA) and in a subgroup of 296 IBS subjects (P < 0.05, one-way ANOVA), there was a significant association between the severity of reported constipation and the presence of methane. The opposite was true for diarrhea (P < 0.001). If a breath test was methane positive, this was 100% associated with constipation predominant IBS. Furthermore, IBS had a greater prevalence of methane production than Crohn's or UC. In fact, methane was almost nonexistent in the predominantly diarrheal conditions of Crohn's and UC. In conclusion, a methane positive breath test is associated with constipation as a symptom. PMID- 12645796 TI - Gelatinase A (MMP-2), collagenase-2 (MMP-8), and laminin-5 gamma2-chain expression in murine inflammatory bowel disease (ulcerative colitis). AB - Dextran sulfate sodium-induced inflammatory bowel disease in mice resembles human ulcerative colitis. In inflammatory bowel diseases matrix metalloproteinases contribute to tissue degradation. Laminin-5 is an anchoring filament protein in the basement membrane area that can be cleaved by matrix metalloproteinases. We investigated the expression of matrix metalloproteinases-2 and -8 and laminin-5 gamma2-chain in dextran sulfate sodium-induced mice by immunohistochemistry and in situ hybridization. Matrix metalloproteinase-8 expression was evidenced in the colon surface epithelial cells and the protein was more abundant in dextran sulfate sodium-induced mice colon. Matrix metallproteinase-2 and laminin-5 gamma2 chain colocalized in the colon surface epithelial cells and in the basement membrane zone as demonstrated by double immunostaining. In dextran sulfate sodium induced colon, matrix metalloproteinase-2 immunoreactivity was detected in epithelial cells in the lower parts of the crypt and surrounding the degraded crypts. Matrix metalloproteinase-2 and -8 could participate in the local epithelial inflammatory processes and tissue destruction. The presence of laminin 5 gamma2-chain indicates alternative anchoring mechanisms in the colon, a compartment devoid of hemidesmosomes. PMID- 12645798 TI - Evidence for a carrier-mediated mechanism for thiamine transport to human jejunal basolateral membrane vesicles. AB - Recent studies from our laboratory have demonstrated the presence of a pH dependent, amiloride-sensitive, electroneutral carrier-mediated exchange for thiamine absorption in the human small intestinal brush-border membrane vesicles. However, the mechanism of thiamine transport across the human small intestinal basolateral membrane is not understood. The present study was aimed to characterize the mechanism of thiamine transport across the basolateral membranes of the human jejunum. Basolateral membrane vesicles (BLMV) were purified from mucosal scrapings of organ donors, utilizing a Percoll continuous density gradient centrifugation technique. The results showed [3H] thiamine uptake into BLMV to be: (1) markedly stimulated in the presence of an outwardly directed H+ gradient (pH 5.5in/7.5out); (2) significantly inhibited by amiloride in a dose dependent manner; (3) sensitive to temperature and medium osmolarity and insensitive to changes in membrane potential; (4) not influenced by the addition of 1 mM Mg(2+)-ATP, inside and outside the vesicles in the presence of Na+ and K+; (5) inhibited by structural analogs-amprolium, oxythiamin, and unlabeled thiamine (100 microM); (6) not affected by organic cations, eg, TEA, N-methyl nicotinamide (NMN), and choline; and (7) saturable as a function of concentration (apparent Km of 0.76 +/- 0.21 microM and a V(max) of 1.38 +/- 0.35 pmol/mg protein/10 sec). These results indicate the presence of a proton gradient dependent specialized carrier-mediated exchange mechanism for thiamine transport across the human jejunum basolateral membranes. PMID- 12645797 TI - Histological and immunological features of appendix in patients with ulcerative colitis. AB - Patients with ulcerative colitis (UC) have a less frequent prior history of appendectomy than the general population. The aim of the present investigation was to elucidate histological and immunological characteristics of the appendix in UC and to assess the effect of appendectomy on the disease. Nine subjects with mildly active UC were treated by surgical appendectomy. In four subjects, the histological findings of the appendix were compatible with ulcerative appendicitis. CD3+CD4+CD25+, CD3+CD4+CD45RO+, and CD3+CD8+CD45RO+ appendiceal mononuclear cells were significantly higher in UC than in acute appendicitis and in normal appendix. There was a trend towards higher mRNA transcripts of IFN gamma in the appendix of UC than those in other two groups. Clinical activity index decreased significantly four weeks after the appendectomy, although the effect was transient. The appendix is a site of involvement in UC, where mononuclear cells are presumed to be at a state of basal activation. PMID- 12645799 TI - Electrogenic ion transport in mammalian colon involves an ammonia-sensitive apical membrane K+ conductance. AB - It is remarkable that high ammonia concentrations can be present within the colonic lumen without compromising normal epithelial function. We investigated the impact of luminal ammonia on Cl- secretion in native tissue. Stripped human colonic mucosa and unstripped rat distal colon were used. Paired samples were mounted in modified Ussing chambers for electrophysiological studies. In rat distal colon, apical ammonia dose-dependently blocked forskolin-activated short circuit current with an IC50 to approximately 5 mM. Basolateral NH4Cl was less effective. Luminal methylamine (50 mM), chromanol 293B (10-50 microM), and Ba2+ (5 mM) blocked cAMP-activated short-circuit current but apical clotrimazole (100 microM) was without effect. In stripped human colonic mucosa, luminal but not basolateral NH4Cl (10 mM) and luminal Ba2+ (5 mM) suppressed forskolin-activated short-circuit current. Ammonia may be an endogenous regulator of colonic water and salt secretion. Apical K+ channels may be involved in the regulation of cAMP stimulated Cl- secretion in mammalian colon. PMID- 12645800 TI - Effect of ribavirin and amantadine on early hepatitis C virus RNA rebound and clearance in serum during daily high-dose interferon. AB - The early rebound in serum HCV RNA during HCV dynamic studies with high-dose interferon may be due to de novo infection with interferon escape quasispecies. We simultaneously measured serum alanine aminotransferase (ALT) and HCV RNA at rapid intervals in chronic HCV liver disease patients during interferon therapy alone or in combination with ribavirin and amantadine. HCV RNA declined rapidly between 0 and 48 hr in all patients (phase 1). Ribavirin and amantadine significantly increased this phase 1 decline. In all four monotherapy patients with viral rebound, the increasing levels of HCV RNA were associated with a parallel increase in serum ALT, consistent with a hepatitis flare or de novo infection. By contrast, in the four monotherapy patients without viral rebound, and all eight patients receiving combination therapy, the slow progressive phase two decay was associated with declining serum ALT levels. Ribavirin or ribavirin and amantadine significantly and incrementally increased the phase two HCV RNA clearance. Dynamic sequencing in the HVR1 region in one rebound patient confirmed the potential for rapid evolutionary changes during interferon therapy. These preliminary data suggest that early viral rebound might be associated with de novo infection with interferon escape HCV variants, which in turn are attenuated by ribavirin and amantadine. PMID- 12645802 TI - Biochemical markers of fibrosis in patients with chronic hepatitis C: a comparison with prothrombin time, platelet count, and age-platelet index. AB - As an alternative to liver biopsy, an index of five biochemical markers (alpha2 macroglobulin, apolipoprotein A1, haptoglobin, total bilirubin, gamma-glutamyl transpeptidase) has been shown to predict the severity of hepatitis C-related fibrosis. The objective of this study was to compare this index with other markers frequently used for this purpose (prothrombin time, platelets, age platelet index). In 323 hepatitis C-infected patients, the discriminative values of these markers for F2-F4 fibrosis (by the METAVIR classification) were compared. By multiple logistic regression analysis, only the five-marker index (P < 0.0001) and prothrombin time (P = 0.02) were independently predictive of F2-F4 fibrosis. For this outcome, the area under the receiver operating characteristic curve was significantly higher for the five-marker index (0.836 +/- 0.024) than the age-platelet index (P = 0.002), and the platelet count and prothrombin time (P < 0.001), indicating greater diagnostic value. The addition of the latter markers to the five-marker index proved unhelpful for increasing its accuracy. In conclusion, an index of five biochemical markers accurately predicts significant hepatitis C-related fibrosis and is superior to traditional markers. PMID- 12645801 TI - Intensive interferon therapy does not increase virological response rates in African Americans with chronic hepatitis C. AB - To determine if an intensive regimen of daily, high-dose interferon would improve the initial response rates to therapy for hepatitis C genotype 1 among African American and Caucasian patients, we conducted a retrospective analysis of a treatment trial conducted between October 1995 and June 1997. Patients were randomized to 24 weeks of therapy with interferon--alpha-2b at either 5 MU daily or 3 MU three times a week. On the standard interferon regimen (3 MU three times a week) African Americans and Caucasians had similar initial response rates. However, unlike Caucasians, African Americans did not have an increased initial virological response when treated with an intensive, daily dose regimen. Levels of HCV RNA decreased more slowly during the first 12 weeks of therapy among African Americans. Nelson-Aalen cumulative hazard estimates for the different race and dose combinations revealed that Caucasians who received daily interferon were most likely to have an initial response (logrank, P < 0.001). PMID- 12645803 TI - Hemodynamic effects of acute and chronic administration of vapreotide in rats with cirrhosis. AB - The aim of this study was to assess the hemodynamic effects of acute and chronic administration of vapreotide, a somatostatin analog, in rats with intrahepatic portal hypertension induced by dimethylnitrosamine (DMNA) administration. Acute effects were evaluated at baseline and 30 min after placebo (N = 13) or vapreotide (8 /microg/kg/hr, N = 13) infusions in DMNA rats. Chronic hemodynamic effects were evaluated using subcutaneous implants for five weeks in anesthetized DMNA rats (placebo: N = 13, vapreotide: N = 13) and in sham rats (placebo: N = 10, vapreotide: N = 10). Hemodynamic measurements included splenorenal shunt blood flow (SRS BF) by the transit time ultrasound (TTU) method and cardiac output by the combined dilution-TTU method. Acute administration of vapreotide significantly decreased SRS BF (-17.3 +/- 19 vs - 1.1 +/- 14%, P < 0.05) and portal pressure (-8 +/- 9 vs 0 +/- 8%, p < 0.05) compared to placebo without systemic effects. Chronic administration of vapreotide significantly reduced the increase in SRS BF (2.4 +/- 1.5 vs 1.2 +/- 1.0 ml/min, P < 0.05) and cardiac index (50 +/- 15 vs 33 +/- 10 ml/min/100 g, P < 0.0001) while portal pressure and blood flow, and mean arterial pressure were not significantly changed compared to placebo. In conclusion, the acute administration of vapreotide decreased collateral circulation blood flow while chronic administration attenuated its development. Vapreotide seems to have a vasoconstrictive effect on collateral circulation. PMID- 12645804 TI - No additive effect between Helicobacter pylori infection and portal hypertensive gastropathy on inducible nitric oxide synthase expression in gastric mucosa of cirrhotic patients. AB - Increased expression of inducible nitric oxide synthase (iNOS) has been reported in gastric mucosa of patients with Helicobacter pylori infection or portal hypertensive gastropathy (PHG) but whether there is an additive or synergistic impact between H. pylori and PHG on iNOS expression was unknown. Sixty cirrhotic patients and 21 age-matched control subjects without liver disease were included. Biopsies from the gastric antrum and body were obtained for quantitative histological assessment and immunohistochemical staining for iNOS. iNOS staining was detected in endothelial cells and macrophages. In the absence of PHG, H. pylori significantly induced iNOS expression in cirrhotic patients. PHG also significantly induced iNOS expression in H. pylori-negative patients. However, there was no synergistic or additive effect between H. pylori and PHG on this expression. Furthermore, expression of iNOS was significantly higher in patients with severe PHG than in those with mild PHG and without PHG. PMID- 12645805 TI - Sarcoidosis after use of interferon for chronic hepatitis C: report of a case and review of the literature. AB - Although interferon has not been classified in the pathogenesis of sarcoidosis, it may rarely lead to this disease during treatment of chronic hepatitis C. The case of a 36-year-old woman with chronic hepatitis C who developed sarcoidosis within 10 weeks of treatment with recombinant interferon-alpha2a and ribavirin is described and all seven similar cases published in English from 1989 to 2001 are discussed. PMID- 12645806 TI - Symptomatic gastrointestinal sarcoidosis: case report and review of the literature. PMID- 12645807 TI - Effects of orthotopic liver transplantation on vasoactive systems and renal function in patients with advanced liver cirrhosis. AB - The effects of Orthotopic liver transplantation (OLT) on renal function and major vasoactive factors was assessed in end-stage cirrhotic patients. Renal function, mean arterial pressure, and plasma vasoactive hormones were measured In 22 cirrhotic patients with refractory ascites before and after OLT. Before OLT, mean arterial pressure, glomerular filtration rate, free water clearance, and fractional sodium excretion serum sodium levels were decreased. In addition, serum creatinine and plasma levels of vasoactive factors were increased. Ten of these patients fulfilled criteria of hepatorenal syndrome (HRS). Nine to 12 months after transplantation, renal function had improved and plasma levels of vasoactive factors had decreased significantly in all patients, including those with HRS. However, glomerular filtration rate remained subnormal and plasma endothelin-1 levels and plasma renin activity remained increased in most of them. In conclusion, OLT improves renal function in patients with end-stage liver cirrhosis, including those with HRS. However, renal function remains subnormal in most of these patients. PMID- 12645809 TI - Liver transplantation for propylthiouracil-induced acute hepatic failure. PMID- 12645808 TI - Syndromic incidence of ovarian carcinoma after liver transplantation, with special reference to anteceding breast cancer. AB - Ovarian cancer is the gynecologic malignancy with the highest number of deaths in the United States. Previous studies had found a decreased incidence of female gynecological malignancies after liver transplantation. In order to estimate the incidence of ovarian carcinoma after liver transplantation, we evaluated 1708 consecutive liver transplant recipients from 1984 to 2001. Of them, 770 (43%) were female. Routine follow-ups were performed at 1, 2, 5, and 10 years after transplantation. There were two cases of ovarian carcinoma. Both occurred in recipients with a previous history of breast cancer. Based on these data, we conclude that the incidence of ovarian cancer is 1:385 among all female liver transplant recipients, and 1:6.5 among those with a history of pretransplant breast cancer. We recommend that regular check-ups should be undertaken, especially in the population at highest risk. PMID- 12645810 TI - Tumor-associated hypoglycemia from metastatic colorectal adenocarcinoma: case report and review of the literature. PMID- 12645813 TI - Chronic exposure to subtoxic levels of peroxidized lipids suppresses mucosal cell turnover in rat small intestine and reversal by glutathione. AB - Oxidative challenge can compromise intestinal growth and death responses. This studyexamines the effect of chronic consumption of subtoxic levels of peroxidized lipids on intestinal redox balance and turnover and the effect of glutathione (GSH) supplementation. Male Sprague-Dawley rats were fed standard rat chow or 4% peroxidized menhaden oil chow (2-8 weeks). Intestinal GSH and glutathione disulfide (GSSG), GSH synthetic and redox enzymes as well as proliferative (ornithine decarboxylase, ODC) and apoptotic activities were evaluated. Chronic peroxide intake did not affect overall animal growth, but decreased intestinal GSH/GSSG ratio that directly correlated with decreased GSH and increased GSSG, and suppressed peak circadian ODC activities and post-prandial mucosal apoptosis. Supplementation with GSH restored the mucosal GSH/GSSG ratio and abrogated the peroxide-induced suppression of intestinal cell turnover. Collectively, the results show that chronic lipid peroxide consumption induces intestinal GSH redox imbalance that interferes with regulation of enterocyte death and proliferation in vivo. These disruptive effects of lipid peroxides were reversed by GSH supplementation in accordance with the normalization of tissue GSH/GSSG redox balance. PMID- 12645812 TI - Frequent Ki-ras mutations and transforming growth factor-alpha expression in adenocarcinomas of the small intestine: report of 7 cases. PMID- 12645811 TI - Perils of immunohistochemistry: variability in staining specificity of commercially available COX-2 antibodies on human colon tissue. AB - We assessed the immunohistochemical (IHC) staining patterns of three commercially available COX-2 antibodies in human tissues. The location of positive stain in sequential serial 4-mu sections of formalin fixed, paraffin embedded tissue differed considerably. Staining patterns ranged from diffuse cytoplasmic to occasional perinuclear staining. Thus, marked variability in staining results from use of different antibodies, making it important to consider the antibody used when comparing reports of COX-2 staining from different investigators. PMID- 12645815 TI - Antiproliferative and proapoptotic effect of ascorbyl stearate in human pancreatic cancer cells: association with decreased expression of insulin-like growth factor 1 receptor. AB - Pancreatic cancer is an aggressive tumor with short median survival and high mortality rate. Alternative therapeutic modalities are currently being evaluated for pancreatic cancer. Here we studied the effects of ascorbyl stearate (Asc-S), a nontoxic, lipophilic derivative of ascorbic acid, on pancreatic cancer. Treatment of human pancreatic carcinoma cells with Asc-S (50-200 microM) resulted in a dose-dependent inhibition of their proliferation. Asc-S slowed down the cell cycle, accumulating, PANC-1 cells in late G2-M phase. Furthermore, Asc-S treatment (150 microM) markedly inhibited growth in soft agar and facilitated apoptosis of PANC-1 cells but not of Capan-2 cells. These effects were accompanied by a significant reduction in insulin-like growth factor 1 receptor (IGF1-R) expression, as compared to untreated controls. Interestingly, Capan-2 cells, the least responsive to Asc-S treatment, did not overexpress the IGF1-R. The results demonstrate the efficacy of Asc-S in inhibing growth of pancreatic cancer cells and warrant additional studies to explore the potential utility of this compound as an alternative and/or adjuvant therapeutic modality for pancreatic cancer. PMID- 12645814 TI - Correlation of Ki-67, p53, and Adnab-9 immunohistochemical staining and ploidy with clinical and histopathologic features of severely dysplastic colorectal adenomas. AB - Variations of Ki-67, p53, and Adnab-9 monoclonal antibody reactions in colonic adenomas may be associated with colonic cancer risk. We studied the predictive value of these markers for adverse behavior in severely dysplastic colorectal adenomas, such as an associated carcinoma, multiplicity of adenomas, and subsequent development of adenomas. For this purpose we compared theclinical, gross, and histologic characteristics of highly dysplastic index polyps in 42 patients with Ki 67, p53, and Adnab-9 immunostaining and other molecular markers. Polyps were removed endoscopically, and severely dysplastic polyps were stained immunohistochemically with Ki-67, Adnab-9, and p53 protein by the avidin biotin conjugate (ABC) technique. Quantitative DNA (QDNA) was analyzed by computer assisted image analysis. Ki-67 immunohistochemistry showed reversal of normal distribution of nuclear staining from the normal basal position to the upper third of the colonic crypts. This abnormality of immunostaining in dysplastic adenomas was the earliest detected by the panel we used. A statistically significant correlation was seen between invasiveness of carcinoma in the index polyp and polyp size (P = 0.003), sessile morphology (P = 0.037), and villous or tubulovillous histology (P = 0.019). In the index adenoma, p53 positivity was correlated with multiplicity at initial examination (P = 0.053), villous histology (P = 0.053), invasiveness of carcinoma (P < 0.003), and recurrence of colorectal adenomas (P = 0.025). Although p53 positivity and aneuploidy were correlated with invasiveness of carcinoma in the index polyp (P = 0.025), Adnab-9 positivity was not. However, Adnab-9 positivity in the index polyp was associated with multiplicity of adenomas (P = 0.04) as well as recurrence of adenomas (P < 0.024). In conclusion, in addition to the morphologic and histologic markers already known, Ki-67, Adnab-9 antibody, and p53 protein may be prognostic indicators useful in follow-up of patients with severely dysplastic colorectal adenomas. Adnab-9 antibody may identify a field defect in above-average-risk adenoma-bearing patients. PMID- 12645816 TI - Modulation of gene expression by genetic and environmental heterogeneity in timing of a developmental milestone. AB - The expression of many important behavioral outcomes is contingent on passing a developmental milestone, such as puberty, or exposure to an age-dependent risk factor, such as alcohol or drugs. Current models for genetic effects on development have not provided explicit theoretical insight about possible patterns of epistasis and genotype x environment interaction generated by genetic and environmental heterogeneity in the timing of developmental milestones that influence gene expression. We explore theoretically the effects of individual differences in the timing of a developmental milestone on the expression of genetic differences in outcome. The consequences of additive genetic variation and shared environmental variation on timing are examined. Additive genetic effects on timing generate transient epistatic effects on outcome because they modulate the expression of otherwise additive genetic differences in outcome. Epistatic effects will be greatest at ages when there is the most heterogeneity in developmental maturity. Epistasis will gradually disappear as proportionately more individuals mature. Similarly, shared environmental effects on the timing of a milestone will generate transient genotype x shared environment interaction appearing as shared environmental effects on outcome that are greatest when there is the most heterogeneity in developmental maturity. The impact of these effects on age patterns in twin correlations is examined and their relevance to current data on developmental change discussed briefly. PMID- 12645817 TI - Association analysis of exonic variants of the GABA(B)-receptor gene and alpha electroencephalogram voltage in normal subjects and alcohol-dependent patients. AB - Based on pharmacologic evidence, it has been suggested that GABA(B) receptors may play a crucial role in the generation of alpha-electroencephalogram (EEG) oscillations. We tested whether three exonic variants of the gene encoding the human GABA(B) receptor (GABA(B)R1) modify scalp-recorded alpha-EEG voltage. One hundred twenty-eight patients suffering from alcoholism and 114 normal subjects were investigated. Alcohol-dependent patients were included because evidence exists that the frequently observed alpha low voltage in these subjects is at least partly a trait variable. Logistic regression analyses revealed no associations between alpha-EEG voltage and polymorphic variations in exon 1a1 or exon 11. A significant interaction was observed for an exon 7 substitution polymorphism and diagnosis (P = 0.009). Post hoc analyses showed an association between EEG phenotype and exon 7 genotype in normal subjects only. It is concluded that this particular association may only be observable under physiologic conditions and that alpha low voltage in alcohol-dependent subjects is under the control of either different genes or that it is related to the disease process. PMID- 12645818 TI - Complex inheritance of male courtship song characters in Drosophila virilis. AB - We analyzed the inheritance of two male courtship song characters, number of pulses in a pulse train, and length of a pulse train in Drosophila virilis. Biometric analyses of song differences among 16 crosses over three generations (parental and reciprocal F1, F2, and backcrosses) were performed using two different approaches. The joint scaling test revealed significant additive and dominance components, and also significant additive interaction between maternal and progeny genotypes. The direction of dominance was toward shorter and denser pulses in D. virilis. In addition, planned comparisons (contrast analyses of variance) between different generations revealed significant Y chromosomal and transient maternal factors and their interactions, with all other factors contributing to differences in the means of song characters. These results suggest that the genetic basis of courtship song characters in D. virilis is primarily of autosomal polygenic nature, with small but significant Y chromosomal and nonchromosomal maternal effects. PMID- 12645819 TI - Role of X chromosomal song genes in the evolution of species-specific courtship songs in Drosophila virilis group species. AB - In the Drosophila virilis group the males of the virilis phylad species produce courtship song consisting of pulse trains with no pauses between successive sound pulses, whereas the males of the montana phylad species produce songs with clear pauses between the sound pulses. We obtained song data for F1 hybrids between D. virilis (representing the virilis phylad) or D. flavomontana (representing the montana phylad) females and the males of several species of the D. virilis group to study the interaction of X chromosomal and autosomal song genes affecting species differences in song. In crosses with D. virilis females, X chromosomal (or maternal) factors masked variation in pulse length despite variation in heterospecific autosomal song genes. To the contrary, in crosses with D. flavomontana females, X chromosomal genes largely determined the pause length and interacted with autosomal genes to determine the pulse length. In the montana phylad species, pulse length showed dominance toward shorter pulses and pause length toward longer pauses. The first-mentioned trait also indicated the epistatic effects of X chromosomal and autosomal components. PMID- 12645820 TI - Quantitative study of the response to genetic selection of the Wistar audiogenic rat strain (WAR). AB - To study the relationship between genetics and epilepsy, Wistar rats susceptible to audiogenic seizure were selected from the main breeding stock of the Ribeirao Preto School of Medicine at the University of Sao Paulo, Brazil, and inbred. The criteria for selection were the highest seizure severity index (SI) and shortest latencies of the first running fit (LI). Because behavioral response to sound stimulation (120 dB) at 70 to 78 days of age was very stable, SI and LI were evaluated within this age range. Analysis of 9450 observations in 1575 animals from the 3rd to 17th generations demonstrated significant effects of generation, parity, and litter on SI and of generation and litter on LI. The SI and LI averages were, respectively, 37.13 and 22.82 s in the 3rd generation and 83.06 and 7.84 in the 17th generation. Heritabilities of both characters were estimated, by maximum likelihood, as 0.37 +/- 0.066 and 0.44 +/- 0.059, respectively. Because a significant regression related individual breeding values for both SI and LI to generation number, we concluded that genetic selection has a positive impact on the traits analyzed. Therefore, the Wistar audiogenic rat (WAR) strain appears, as per the 17th to 20th generations of genetic selection to be an audiogenic rat strain suitable for epilepsy studies. PMID- 12645821 TI - Influence of shock training and explicit fear-conditioned cues on sleep architecture in mice: strain comparison. AB - Fear conditioning is thought to model anticipatory anxiety. Inbred mouse strains exhibit different levels of reactivity to aversive environmental stimuli, which may reflect anxiety. We examined the effects of fear conditioning on sleep in mouse strains that differ on behavioral measures of anxiety. Mice (BALB/cJ [C], C57BL/6J [B6], CB6F1/J [CB6], n = 7-10 per strain) were implanted with transmitters for recording sleep by telemetry. Baseline sleep was recorded, and the mice were trained to associate a cue (tone) with footshock (15 cue-shock pairings on 4 consecutive days). Sleep was recorded after shock training and again 4 to 5 days later after presentation of the cue alone. Shock training produced a relatively selective suppression of rapid eye movement sleep (REM) that was greater in the C strain compared to the B6 and CB6 mice. Post-training exposure to the cue alone suppressed REM in all strains. The C strain exhibited a relatively greater immediate suppression of REM, and the CB6 hybrid mice showed the greatest overall suppression of REM. These data demonstrate that stimuli associated with an aversive event can alter sleep and suppress REM in much the same way as exposure to the event itself. Fear conditioning may provide a model for examining genetic and neural mechanisms underlying the influence of anxiety on sleep. PMID- 12645822 TI - Motor coordination, exploration, and spatial learning in a natural mouse mutation (nervous) with Purkinje cell degeneration. AB - The nervous mouse mutation causes a relatively selective degeneration of Purkinje cells in the cerebellar cortex. The mutants were compared to age-matched controls of the same background strain in tests of motor activity and coordination, spontaneous alternation, and spatial learning in the Morris water maze. As expected from their ataxia, the nervous mutants were impaired in stationary beam, coat-hanger, and rotorod tests of motor coordination. The nervous mutants were also impaired in the submerged but not in the visible platform condition of the Morris water maze, attributable to a spatial deficit, and displayed a higher level of motor activity in an automated chamber. The deficit in spontaneous alternation rates seen in nervous mutants is accountable by reduced motivation, disinhibition, or spatial disorientation. PMID- 12645823 TI - DNA from buccal swabs recruited by mail: evaluation of storage effects on long term stability and suitability for multiplex polymerase chain reaction genotyping. AB - We provide details of an inexpensive and rapid method for extraction of DNA from buccal swabs (including samples received through the mail) and from a range of other tissue samples. The procedure we have developed provides amounts of DNA adequate for several thousand polymerase chain reactions (PCRs), and we have validated its potential for long-term storage. Samples stored for > 4 years are of comparable concentration and provide as robust PCR templates as those tested immediately after extraction. The availability of this technology is of considerable significance in planning DNA banks from population collections and cohorts. PMID- 12645825 TI - Breathing retraining and exercise conditioning in patients with chronic obstructive pulmonary disease (COPD): a physiological approach. AB - In this review we shall consider the commonest techniques to reduce dyspnea that are being applied to patients with chronic obstructive pulmonary disease (COPD) subjected to a pulmonary rehabilitation program (PRP). Pursed lip breathing (PLB) and diaphragmatic breathing (DB) are breathing retraining strategies employed by COPD patients in order to relieve and control dyspnea. However, the effectiveness of PLB in reducing dyspnoea is controversial. Moreover, DB may be associated with asynchronous and paradoxical breathing movements, reflecting a decrease in the efficiency ofthe diaphragm. Exercise training (EXT) is a mandatory component of PRP.EXT has been shown to improve exercise performances and peripheral muscle strength. Recent studies have focused on the effect of EXT on breathlessness. However, concerns persist as to whether the decreased sensation of dyspnea for a given exercise stimulus is principally due to psychological benefits of rehabilitation or to improved physiological ability to perform exercise. The effect of EXT on breathlessness may be reinforced by inhaling oxygen. However, two studies have recently shown that breathing supplemental oxygen during training has either a marginal effect or no advantage over training. In a comprehensive PRP, strength training (ST) and arm endurance training (AET) could have a role in decreasing peripheral muscle weakness and metabolic and ventilatory requirements for AET. The role of unloading the respiratory muscles during EXT has to be PMID- 12645824 TI - Genotyping of three candidate genes after whole-genome preamplification of DNA collected from buccal cells. AB - The amount of genomic DNA obtained from buccal cell methods may be suboptimal for large-scale genetics projects, because the quantity of DNA may be insufficient for the number of analyses proposed. Primer extension preamplification (PEP) methods that can amplify the entire genome 100-fold or more, offer a potential solution to this problem. We compared PEP buccal DNA with genomic buccal DNA from 315 individuals from 97 families of the Colorado Longitudinal Twin Study for three loci: the dopamine transporter, dopamine D4 receptor, and serotonin transporter. A total of 1890 genomic and 1890 PEP alleles were assessed, and 1670 comparisons (88%) agreed after a single determination. Fifty-three individuals had one or more failed initial polymerase chain reactions (PCR), with 81 failed PCRs in total, accounting for 162 missing allele calls. The failed PCRs were repeated once, and 146 of the missing allele calls were recovered. Comparisons between genomic and PEP DNA allele calls showed 37 individuals had one or more discrepancies, for a total of 52 inconsistencies. Of these, the initial PEP result was found to be correct in 18 cases, the initial genomic result was found to be correct in 25 cases, and 9 could not be resolved. Overall, rates of true calls, missing data, and genotyping errors for genomic and PEP DNA samples were nearly identical: of the 1890 genotypes assessed, true calls were found in 1845 genomic and 1840 PEP samples, missing genotypes in 18 genomic and 16 PEP samples, and incorrect assignments in 18 genomic and 25 PEP samples. These results suggest that routine whole-genome preamplification of genomic DNA is an appropriate method for providing DNA to genotype these loci. PMID- 12645826 TI - Have guidelines for the management of community-acquired pneumonia influenced outcomes? PMID- 12645827 TI - Cost-effectiveness of fluticasone propionate in the treatment of chronic obstructive pulmonary disease: a double-blind randomized, placebo-controlled trial. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a debilitating disease and places a large financial burden on health-care systems and society. We prospectively evaluated the cost-effectiveness offluticasone propionate (FP) treatment in patients with moderate-to-severe COPD, who were symptomatic on regular bronchodilator therapy. METHODS: An economic analysis was performed in a 6-month, randomized, double-blind clinical trial comparing FP 1,000 microg/day with placebo in 281 patients aged 45-79 years with symptomatic moderate-to-severe COPD. Data on clinical efficacy, health-care resource use and productivity loss associated with the management of COPD were prospectively collected. The main outcome measures were the incremental cost-effectiveness of achieving a > or = 10% improvement in FEV1 and of remaining exacerbation-free throughout the study. The economic evaluation was costed from the perspective of the NHS (direct costs) and of society (direct and indirect costs). RESULTS: FP was significantly more effective than placebo in terms of the proportions of patients demonstrating a > or = 10% improvement in FEV1 (32 vs. 19%; P = 0.02) and remaining free of moderate/severe exacerbations (75 vs. 63%; P = 0.02). The difference between the groups in total costs was not significantly different. Incremental cost effectiveness analyses showed that the additional clinical benefits of FP relativeto placebo, in terms of a > or = 10% improvement in FEV1 or an increased number of patients free of moderate/severe exacerbations, were achieved at minimal additional costs from an NHS perspective (additional 0.25 pounds per day for bath) or at a net saving from a societal perspective. Sensitivity analysis showed that these results were robust to changes in the underlying assumptions. CONCLUSIONS: Treatment with FP was associated with statistically significant clinical benefits in patients with moderate-to-severe COPD currently symptomatic on regular bronchodilator therapy. As the differences in direct and total costs compared with placebo were small and non-significant, this treatment can be considered cost-effective in this patient population. PMID- 12645828 TI - Perception of dyspnea during exercise-induced bronchoconstriction. AB - After strenuous physical exercise, many subjects show a significant bronchoconstriction and report dyspnea. Despite this clinical condition being a commonly encountered situation during daily life, which may be responsible for substantial disability there is little information on the relationship between the perception of dyspnea and exercise-induced bronchoconstriction (EIB) after a standardized exercise challenge. For these reasons, we evaluated 200 consecutive outpatients (median age 13 years, ranging from 5 to 56 years) referred to our laboratory to perform an exercise test out of suspicion of EIB. On exercise challenge, perception of dyspnea was rated on a modified bipolar Borg scale immediately before each FEV1 measurement. Sixty-nine (35%) subjects had a positive exercise challenge, defined as a decrease of at least 20% in FEV1 from baseline. Both the onset and the decay of dyspnea preceded those of bronchoconstriction. Overall, the rating of dyspnea in the laboratory was well related with the reports of exercise-related symptoms. Similarly, 36 of 77 (47%) asthmatics with a history of exertional symptoms and 24 of 65 patients (40%) without a history had a positive challenge. Asthmatics reporting exertional symptoms perceived a greater magnitude of dyspnea after exercise independently from the degree of bronchoconstriction. Overall, dyspnea was significantly but loosely correlated to the magnitude of decrease in FEV1, being also influenced by age, gender and BMI. CONCLUSIONS: We conclude that dyspnea recorded in the laboratory after exercise test is related to exertional symptoms reported during real life, but not completely related to EIB. The rating of dyspnea is a well suited model to study naturally occurring exercise-induced dyspnea and a useful tool to enlarge the results of an exercise challenge. PMID- 12645829 TI - Regulation of protease-activated receptor-1 in mononuclear cells by neutrophil proteases. AB - Neutrophils and mononuclear cells are implicated in the pathogenesis of several inflammatory conditions including chronic obstructive pulmonary disease (COPD). Neutrophil-derived serine proteases, such as cathepsin G (CG) and neutrophil elastase (NE), may interact with mononuclear cells via protease-activated receptors (PARs) which are seven-transmembrane G protein-coupled receptors activated by proteolytic cleavage of the extracellular N-terminus, and which, on activation, induce the release of several mediators and cytokines. We determined whether CG and NE could affect PAR-1 expression and function in mononuclear cells. Human blood mononuclear cells were isolated from 20 healthy donors. Surface and intracellular receptor expression and calcium mobilisation (using the calcium chelator, FLUO3-AM) were studied by fluorescence-assisted cell sorting (FACS analysis). Positive controls, i.e. thrombin (0.1-100 mU/ml) and the PAR-1 activating peptide SFLLRN (100 microM) induced a rapid and transient intemalisation of PAR-1 in monocytes and lymphocytes. CG but not NE had a similar effect. By contrast, in monocytes intracellular calcium mobilisation was induced by thrombin and SFLLRN but not by CG and NE. Thus, CG can induce intracellular PAR-1 sequestration without activation of the receptor, and may act as an antagonist and prevent subsequent activation of PAR-1 in mononuclear cells. These findings may be of relevance to the pathogenesis of COPD. PMID- 12645830 TI - The salmeterol/fluticasone combination is more effective than fluticasone plus oral montelukast in asthma. AB - The aim of this study was to compare the efficacy and safety of salmeterol/fluticasone propionate combination product (SFC) with fluticasone propionate (FP) plus oral montelukast (M) over 12 weeks in symptomatic asthma patients. The study was a multinational, randomised, double-blind, double-dummy, parallel-group design in patients aged > or = 15 years. After a 4-week run-in during which all patients received FP 100 microg twice daily, patients were randomised to inhaled SFC (50/100 microg) twice daily or inhaled FP 100 microg twice daily and oral M 10 mg once daily. Patients kept daily records of their peak expiratory flow (PEF) symptom scores and use of rescue medication. Over the 12-week treatment period, the adjusted increase in mean morning PEF was significantly greater in the SFC group (36 l/min) than the FP/M group (19 l/min; P < 0.001). The improvement in FEV1 was also significantly greater in the SFC group (mean treatment difference 0.11 l; P < 0.001). SFC provided significantly better control of daytime and night-time symptoms and there were fewer exacerbations. Patients in the SFC group were also significantly more likely to have a rescue-free day. Both treatments were equally well tolerated. Combination therapy with FP plus salmeterol (SFC) produced significantly greater improvements in lung function and asthma control than the addition of montelukastto FP. PMID- 12645831 TI - Oral gemifloxacin once daily for 5 days compared with sequential therapy with i.v. ceftriaxone/oral cefuroxime (maximum of 10 days) in the treatment of hospitalized patients with acute exacerbations of chronic bronchitis. AB - In a randomized, open-label, controlled, multicentre study, the clinical and bacteriological efficacy, safety and tolerability of oral gemifloxacin (320 mg once daily, 5 days) was compared with sequential intravenous (i.v.) ceftriaxone (1 g once daily, maximum 3 days) followed by oral cefuroxime axetil (500 mg twice daily, maximum 7 days) in adult hospitalized patients with acute exacerbations of chronic bronchitis (AECB) (n = 274). The clinical success rates at follow-up (21 28 days post-therapy) in the clinical per-protocol population (the primary endpoint) were 86.8% (105/121) for gemifloxacin vs. 81.3% (91/112) for ceftriaxone/cefuroxime (treatment difference = 5.5,95% CI -3.9,14.9). The corresponding clinical results in the clinical intention-to-treat (ITT) population were 82.6% (114/138) vs. 72.1% (98/136), respectively (treatment difference = 10.5,95% CI 0.7, 20.4).Thus, gemifloxacin had significantly higher clinical success rates than ceftriaxone/cefuroxime. The median time to discharge was 9 days in the gemifloxacin group vs. 11 days in the ceftriaxone/cefuroxime group (P = 0.04, Wilcoxon test). At follow-up, 120/138 (87.0%) gemifloxacin treated patients had been discharged from hospital, compared with 111/136 (81.6%) ceftriaxone/cefuroxime-treated patients in the clinical ITT population. Both treatments were generally well tolerated and there was no significant difference between the treatment groups in the incidence or type of adverse events reported. A 5-day course of oral gemifloxacin was shown by this study to be at least equivalent to sequential i.v. ceftriaxone/cefuroxime axetil (for up to 10 days) in patients with AECB who require hospital treatment. PMID- 12645832 TI - Anti-inflammatory activity of 1.8-cineol (eucalyptol) in bronchial asthma: a double-blind placebo-controlled trial. AB - Airway hypersecretion is mediated by increased release of inflammatory mediators and can be improved by inhibition of mediator production. We have recently reported that 1.8-cineol (eucalyptol) which is known as the major monoterpene of eucalyptus oil suppressed arachidonic acid metabolism and cytokine production in human monocytes. Therefore, the aim of this study was to evaluate the anti inflammatory efficacy of 1.8-cineol by determining its prednisolone equivalent potency in patients with severe asthma. Thirty-two patients with steroid dependent bronchial asthma were enrolled in a double-blind, placebo-controlled trial. After determining the effective oral steroid dosage during a 2 month run in phase, subjects were randomly allocated to receive either 200 mg 1.8-cineol t. i.d. or placebo in small gut soluble capsules for 12 weeks. Oral glucocorticosteroids were reduced by 2.5 mg increments every 3 weeks. The primary end point of this investigation was to establish the oral glucocorticosteroid sparing capacity of 1.8-cineol in severe asthma. Reductions in daily prednisolone dosage of 36% with active treatment (range 2.5-10 mg, mean: 3.75 mg) vs. a decrease of only 7% (2.5-5 mg, mean: 0.91 mg) in the placebo group (P = 0.006) were tolerated. Twelve of 16 cineol vs. four out of 16 placebo patients achieved a reduction of oral steroids (P = 0.012). Long-term systemic therapy with 1.8 cineol has asignificant steroid-saving effect in steroid-depending asthma. This is the first evidence suggesting an anti-inflammatory activity of the monoterpene 1.8-cineol in asthma and a new rational for its use as mucolytic agent in upper and lower airway diseases. PMID- 12645833 TI - Nontuberculous mycobacteria: incidence in Southwest Ireland from 1987 to 2000. AB - SETTING: The Southwest of Ireland (Counties Cork and Kerry) 1987-2000, average population 549,500. OBJECTIVE: Nontuberculous mycobacteria (NTM) cause significant morbidity worldwide and the study of epidemiology and characteristics helps in their prevention and treatment. This study was performed to determine the incidence of NTM disease in comparison to Mycobacterium tuberculosis (M. tuberculosis) and Mycobacterium bovis (M. bovis) in Southwest Ireland, over the above time period. DESIGN: A retrospective study was carried out in all human isolates of NTM, M. tuberculosis and M. bovis between 1987 and 2000, in the Southwest Region of Ireland. RESULTS: The mean incidence of NTM (0.4/100,000 population) has risen since 1995, principally of pulmonary Mycobacterium avium intracellulare complex (MAC). The annual incidence of M. tuberculosis in humans over 14 years in the same region was 971/100,000 population with a significant reduction since 1994 and M. bovis remained constant at 0.5/100,000 population. CONCLUSION: The increasing incidence of disease causing NTM noted in Southwest Ireland reflects global data and is surmised to be due to an ageing population, increased incidence related to chronic fibrotic lung disease, and environmental mycobacterial factors. PMID- 12645834 TI - Is there any association between inhaled ipratropium and mortality in patients with COPD and asthma? AB - BACKGROUND: To determine whether ipratropium was associated with premature death in patients with asthma and chronic obstructive pulmonary disease (COPD). METHODS: A longitudinal cohort of 827 patients with COPD and 273 with asthma who were evaluated for compensation by two specialists in pulmonary medicine. RESULTS: By June 1999, 538 of the patients with COPD and 77 of those with asthma had died. Atthe consultation, 77% ofthe COPD patients and 8.1% of the asthmatic patients were treated with inhaled ipratropium. Ipratropium was associated with mortality risk ratio (RR) of 2.0 (95% confidence interval: 1.5-2.6) for COPD and 3.6 (1.8-7.1) for asthma patients. After adjustment for confounding factors [forced expiratory volume 1 s (FEV1), smoking habits, asthma medication, and presence of cor pulmonale] the RR for COPD was 1.6 (1.2-2.1) and for asthma 24 (1.2-5.0). CONCLUSIONS: The increased risk of premature death associated with ipratropium is of concern and necessitates further evaluation, e.g., in a randomised study. PMID- 12645835 TI - Factors associated to lifestyle and risk of adult onset asthma. AB - BACKGROUND: Asthma prevalence has been increasing especially in developed countries. The change seems to be associated with changes in lifestyle. We have made a prospective study to assess the effect of lifestyle factors, including smoking, educational level, physical activity and obesity on adult onset asthma. METHODS: A population of 10,597 adult twins, initially free of asthma was followed for 9 years. The main outcome measure was questionnaire-based report of physician diagnosed asthma. Logistic regression was used to estimate the risk of asthma predicted by lifestyle factors, with adjustment for atopy and respiratory symptoms. RESULTS: Obesity at baseline increased asthma risk (multivariable adjusted OR = 3.00, 95% CI: 1.64-5.50 for those with BMI > or = 30 compared to those with normal weight BMI: 20-24.99). Taller height was associated to lower asthma incidence. Leisure time physical activity had a slightly protective effect on asthma risk among men (P for trend = 0.037) while smoking and education did not have significant effects on the risk of adult onset asthma. CONCLUSIONS: Obesity was associated to the risk of adult onset asthma, while short height and low leisure time physical activity can be considered as other potential risk factors. PMID- 12645836 TI - Lack of evidence for a role of Epstein-Barr virus in the increase of lung cancer in idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) is known as an independent risk factor for lung cancer. Because Epstein-Barr virus (EBV) may be involved in the genesis of IPF as well as certain malignancies, we investigated whether EBV contributes to the increased incidence of lung cancer in IPF The formalin-fixed and paraffin embedded lung sections were prepared from 22 lung cancer patients with IPF and 22 lung cancer patients without IPF All ofthe IPF patients pathologically showed usual interstitial pneumonia. In situ hybridization for EBV-encoded small non polyadenylated RNAs failed to show positive signals in the cancer tissues of either IPF or non-IPF patients. This study did not provide evidence for an etiologic role of EBV in the development of lung cancer in IPF. PMID- 12645837 TI - Peak inspiratory flow through diskus and turbuhaler, measured by means of a peak inspiratory flow meter (In-Check DIAL). AB - With a handheld peak inspiratory flow (PIF) meter (In-Check DIAL), the internal resistance of the Diskus (DKR) and the Turbuhaler (THR) can be simulated by means of calibrated resistances. This study investigated whether patients with asthma or chronic obstructive pulmonary disease could generate a PIF which is optimal for the Diskus (DK) (30 l/min) and theTurbuhaler (TH) (60 l/min). Peak expiratory flow (PEF) and maximal inspiratory pressure (MIP) were also assessed. All 50 patients (30 males, mean age 56.9) achieved the required flow of 30 l/min with the DKR, while six patients (five females) could not generate the optimal flow of 60 l/min via theTHR. Analysis showed that MIP was an independent predictor of an adequate PIF through theTHR. During the study eight patients were treated for an exacerbation of which four could not generate an adequate PIF via the THR. Analysis showed that the female gender and an exacerbation appeared to be independent predictors of the inability to generate an adequate PIF via the THR. Twelve per cent of patients (25% of females) could not generate the optimal inspiratory flow via the THR. When in doubt whether the patient can generate an adequate inspiration, measurement with the In-Check DIAL is recommended. PMID- 12645838 TI - Re: Symptomatic asthma: attendance and prescribing in general practice (Respir Med 96: 102-109): Critique of Nolan and White. PMID- 12645839 TI - Trends in aging--United States and worldwide. AB - The median age of the world's population is increasing because of a decline in fertility and a 20-year increase in the average life span during the second half of the 20th century. These factors, combined with elevated fertility in many countries during the 2 decades after World War II (i.e., the "Baby Boom"), will result in increased numbers of persons aged > or = 65 years during 2010-2030. Worldwide, the average life span is expected to extend another 10 years by 2050. The growing number of older adults increases demands on the public health system and on medical and social services. Chronic diseases, which affect older adults disproportionately, contribute to disability, diminish quality of life, and increased health- and long-term-care costs. Increased life expectancy reflects, in part, the success of public health interventions, but public health programs must now respond to the challenges created by this achievement, including the growing burden of chronic illnesses, injuries, and disabilities and increasing concerns about future caregiving and healthcare costs. This report presents data from the U.S. Bureau of the Census, the World Health Organization, and the United Nations on U.S. and global trends in aging, including demographic and epidemiologic transitions, increasing medical and social costs related to aging, and the implications for public health. PMID- 12645840 TI - Outbreak of group A streptococcal pneumonia among Marine Corps recruits- California, November 1-December 20, 2002. AB - During November 1-December 20, 2002, a total of 163 Marine Corps personnel from the Marine Corps Recruit Depot (MCRD) in San Diego, California, including 160 new recruits, were admitted to the Naval Medical Center San Diego (NMCSD) for possible pneumonia. For 128 (79%) patients, pneumonia was confirmed by chest radiograph; of these 128 cases, 31 (24%) were definitely or probably caused by group A streptococci (GAS). This is the first outbreak of serious GAS-associated illness at a San Diego military training facility since the 1987 outbreak of rheumatic fever and the largest outbreak of GAS pneumonia in the United States since 1968. This report summarizes the results of the investigation of this outbreak, which indicate that GAS infection can occur among military recruit populations despite routine chemoprophylaxis administered to incoming recruits. Instituting routine surveillance for noninvasive GAS disease in military training facilities might prevent future invasive GAS outbreaks. PMID- 12645841 TI - Increase in coccidioidomycosis--Arizona, 1998-2001. AB - Coccidioidomycosis is a systemic infection caused by inhalation of airborne spores from Coccidioides immitis, a fungus found in soil in the southwestern United States and in parts of Mexico and Central and South America. Infection occurs usually following activities or natural events that disrupt the soil, resulting in aerosolization of the fungal arthrospores. Clinical manifestations occur in 40% of infected persons and range from an influenza-like illness (ILI) to severe pneumonia and, rarely, extrapulmonary disseminated disease. Persons at higher risk for disseminated disease include blacks, Filipinos, pregnant women in their third trimester, and immunocompromised persons. During 2001, the Arizona Department of Health Services (ADHS) reported a coccidioidomycosis incidence of 43 cases per 100,000 population, representing an increase of 186% since 1995. To characterize this increase, CDC analyzed data from the National Electronic Telecommunications System for Surveillance (NETSS) and the Arizona Hospital Discharge Database (AHDD), and environmental and climatic data, and conducted a cohort study of a random sample of patients with coccidioidomycosis. This report summarizes the findings of this investigation, which indicate that the recent Arizona coccidioidomycosis epidemic is attributed to seasonal peaks in incidence that probably are related to climate. Healthcare providers in Arizona should be aware that peak periods of coccidioidomycosis incidence occur during the winter and should consider testing patients with ILI. PMID- 12645842 TI - Reconsolidation reconsidered. AB - Some of the considerations that led to a consolidation interpretation of retrograde amnesia (RA), which states that RA results from the disruption of memory processing and storage when neural activity is interrupted by a brain insult, are reviewed here. The time-dependent gradient of memory loss (i.e., new memories are more vulnerable to amnesia than old memories) that characterizes RA seemed to fit nicely with the notion of a cascade of cellular events occurring during the immediate post-acquisition period that would transform a labile representation into a more stable form (i.e., consolidate the memory). However, a variety of observations came to challenge the storage-disruption model, and among these was the finding of amnesia for old but reactivated memories. A recent study by Nader, Schafe, and LeDoux (2000) provides an important analytic extension of the work on "reconsolidation" by showing that inhibition of protein synthesis in the lateral and basal nuclei of the amygdala immediately following the reactivation of old memory will induce retrograde amnesia. We offer a retrieval oriented conceptualization to account for the temporal gradient and the "reconsolidation" phenomena. PMID- 12645843 TI - Blocking in rabbit eyeblink conditioning is not due to learned inattention: indirect support for an error correction mechanism of blocking. AB - Blocking is a classical conditioning task in which prior training to one cue such as a tone reduces learning about a second cue such as a light, when subsequently trained as a tone-light compound. Blocking has been theorized to come about through a US-modulated error correction mechanism by Rescorla & Wagner (1972) as well as through a mechanism of learned inattention as theorized by Mackintosh (1973). In the case of eyeblink conditioning, an error correction mechanism has been hypothesized to take place in the cerebellum while some form of inattention has been hypothesized to take place in the hippocampal region. The hypothesis we are testing is whether the mechanism of learned inattention is involved in blocking in rabbit eyeblink conditioning. If blocking in eyeblink conditioning is produced by a mechanism of learned inattention, then training to a previously blocked cue should be slower than training to that cue in a naive animal. Rabbits that had received tone training followed by tone-light training exhibited blocking. Rabbits that had been previously blocked to the light acquired conditioned responses to the light at the same rate as naive rabbits. This finding failed to support the hypothesis that blocking in rabbit eyeblink conditioning is due to learned inattention, but does support the Rescorla-Wagner mechanism of error correction. The present finding along with previous work on error correction mechanism in the cerebellar-brainstem circuit (Kim et al., 1998) lend support to the theory that blocking, at least in rabbit eyeblink conditioning, seems to be due to an error correction mechanism rather than a learned inattention mechanism. PMID- 12645844 TI - Subcellular interactions between parallel fibre and climbing fibre signals in Purkinje cells predict sensitivity of classical conditioning to interstimulus interval. AB - Classical conditioning of the nictitating membrane response requires a specific temporal interval between conditioned stimulus and unconditioned stimulus, and produces an increase in Protein Kinase C (PKC) activation in Purkinje cells. To evaluate whether biochemical interactions within the Purkinje cell may explain the temporal sensitivity, a model of PKC activation by Ca2+, diacylglycerol (DAG), and arachidonic acid (AA) is developed. Ca2+ elevation is due to CF stimulation and IP3 induced Ca2+ release (IICR). DAG and IP3 result from PF stimulation, while AA results from phospholipase A2 (PLA2). Simulations predict increased PKC activation when PF stimulation precedes CF stimulation by 0.1 to 3 s. The sensitivity of IICR to the temporal relation between PF and CF stimulation, together with the buffering system of Purkinje cells, significantly contribute to the temporal sensitivity. PMID- 12645846 TI - Fractal dynamics of heart beat interval fluctuations in corticotropin-releasing factor receptor subtype 2 deficient mice. AB - Non-linear fractal analysis of cardiac interbeat time series was performed in corticotropin-releasing factor receptor subtype 2 (CRFR2) deficient mice. Heart rate dynamics in mice constitutes a self-similar, scale-invariant, random fractal process with persistent intrinsic long-range correlations and inverse power-law properties. We hypothesized that the sustained tachycardic response elicited by intraperitoneal (ip) injection of human/rat CRF (h/rCRF) is mediated by CRFR2. In wildtype control animals, heart rate was increased to about maximum levels (approximately 750 bpm) while in CRFR2-deficient animals baseline values were retained (approximately 580 bpm). The tachycardic response elicited by ip application is mediated by CRFR2 and is interpreted to result from sympathetic stimulation. However, the functional integrity of CRFR2 would not present a prerequisite to maintaining the responsiveness and resiliency of cardiac control to external environmental perturbations experimentally induced by extrinsic ip application of h/rCRF or under physiological conditions that may be associated with an increased peripheral release of CRF. Under stressful physiological conditions achieved by novelty exposure, CRFR2 is not involved in the cardiodynamic regulation to external short-term stress. While the hypothesis of involvement of CRFR2 in cardiac regulation upon pharmacological stimulation cannot be rejected, the present findings suggest that the mechanism of action is by sympathetic stimulation, but would not unambiguously allow to draw any conclusions as to the physiological role of CRFR2 in the control of cardiac dynamics. PMID- 12645847 TI - Smallpox vaccination: a shot in the dark? PMID- 12645848 TI - Vision examinations for all children entering public school--the new Kentucky law. AB - BACKGROUND: Kentucky was the first state in the United States to pass a law requiring an eye examination by an optometrist or ophthalmologist for each child entering public school, public preschool, or Head Start program for the first time. The law became effective on July 15, 2000. METHOD: Forty-three of 334 Kentucky Optometric Association members were surveyed by the Kentucky Optometric Association. They practiced in 37 of 120 counties throughout Kentucky. Eye examinations for 5,316 children entering the Kentucky school system for the first time were reviewed. The children were divided into groups of 3-year-olds, 4-year olds, 5-year-olds, and 6-year-olds and older. The survey summarized data collected during the period of July 15, 2000 through April 1, 2001. RESULTS: Based on the survey of the clinical assessments of 5,316 eye examinations, a total of 740 children were prescribed spectacle lenses, 181 were diagnosed with amblyopia, 123 children were diagnosed with strabismus, and 44 were diagnosed with other eye diseases. Children in the 6-years-old and above age group were statistically prescribed more spectacle prescriptions than were children ages 3, 4, or 5 years of age. The number of spectacle lens prescriptions, strabismus, amblyopia, and eye diseases diagnosed was independent of county income levels. CONCLUSION: This survey of children entering the Kentucky public school system for the first time showed that 13.92% of the children were prescribed spectacle lenses, 3.40% were diagnosed with amblyopia, and 2.31% were diagnosed with strabismus. PMID- 12645845 TI - Blockade of GABAA receptors in the interpositus nucleus modulates expression of conditioned excitation but not conditioned inhibition of the eyeblink response. AB - The cerebellum and related brainstem structures are essential for excitatory eyeblink conditioning. Recent evidence indicates that the cerebellar interpositus and lateral pontine nuclei may also play critical roles in conditioned inhibition (CI) of the eyeblink response. The current study examined the role of GABAergic inhibition of the interpositus nucleus in retention of CI. Male Long-Evans rats were implanted with a cannula positioned just above or in the anterior interpositus nucleus before training. The rats were trained with two different tones and a light as conditioned stimuli, and a periorbital shock as the unconditioned stimulus. CI training consisted of four phases: 1) excitatory conditioning (8 kHz tone paired with shock); 2) feature-negative discrimination (2 kHz tone paired with shock or 2 kHz tone concurrent with light); 3) summation test (8 kHz tone or 8 kHz tone concurrent with light); and 4) retardation test (light paired with shock). After reaching a criterion level of performance on the feature-negative discrimination (40% discrimination), 0.5 microl picrotoxin (a GABAA receptor antagonist) was infused at one of four concentrations, each concentration infused during separate test sessions. Picrotoxin transiently impaired conditioned responses during trials with the excitatory stimulus (tone) in a dose-dependent manner, but did not significantly impact responding to the inhibitory compound stimulus (tone-light). The results suggest that expression of conditioned inhibition of the eyeblink conditioned response does not require GABAergic inhibition of neurons in the anterior interpositus nucleus. PMID- 12645849 TI - Blurred vision during airline flight reveals prolactinoma. AB - BACKGROUND: Pituitary adenomas can manifest with a variety of endocrinologic signs and symptoms, including amenorrhea, galactorrhea, infertility, and acromegaly. Because of the anatomic location of the pituitary gland, and its proximity to the optic chiasm and cavernous sinuses, pituitary adenomas can also result in decreased visual acuity, diplopia, ophthalmoplegia, visual-field loss, and optic atrophy. In general, these tumors are slow-growing. However, there are reports in the medical literature of patients with previously undiagnosed brain tumors in whom neurological signs suddenly developed when in higher altitudes. CASE REPORT: A 47-year-old woman came in for an evaluation of a one-month history of blurry peripheral vision that occurred during-then persisted following--an international flight. Examination and automated visual-field testing revealed a decrease in her best-corrected visual acuity and a bi-temporal hemianopsia. Subsequent examinations by a neurologist and endocrinologist revealed a significant pituitary adenoma-specifically, a prolactinoma. The patient was treated with bromocriptine and has shown a rapid improvement in her visual field and a regression of the tumor, as evidenced by a repeat MRI. CONCLUSION: In this case, the sudden development of the patients symptoms during an airline flight, and the persistence of the symptoms after landing, resulted in the discovery of a prolactinoma. PMID- 12645850 TI - The reliability of frequency-doubling technology (FDT) perimetry in a pediatric population. AB - PURPOSE: Frequency-doubling Technology IFDT) Perimetry was introduced as a rapid, easy method for detection of visual-field defects in adults. Its reliability, howev, has never been tested among pediatric patients. The purpose of this study was to determine if children could complete a screening program of FDT Perimetry reliably and to define the age range over which it might be most applicable. METHODS: Two hundred fifty-nine normal children (mean age, 10.7 years; range, 4 to 17 years) were asked to complete the FDT C-20-1 screening test as part of their general eye examination. The visual-field results were analyzed for reliability using the number of false-positive errors, fixation losses, and visual-field defects. For the purposes of this analysis, field results were considered unreliable with false-positives > or = 1 or fixation errors > or = 1. The visual-field test was also considered unreliable if > 2 sectors were flagged. RESULTS: Data from 254 children were included in the analysis. Five children were excluded because of suspected malingering or ocular health diagnosis that may have produced a visual-field defect. False-positives were less than one for all but the youngest age group 14 years old). Fixation errors decreed with increasing age and fell below one (became reliable) at approximately 9 years of age, and remained less than one through 17 year. Visual-field loss for purposes of this study when present in more than two sectors was considered unreliable in this normal population. CONCLUSIONS: Children ages 10 years of age and older can reliably complete the FDT C-20-1 screening field test using the strictest criteria, combining false-positives, fixation losses, and visual-field losses. These data are derived from normal subjects, who on complete eye examination, had no ocular disease or reason to suspect visual-field defects. PMID- 12645851 TI - Identifying sources of verification errors in progressive addition lenses. AB - BACKGROUND: Occasional disagreement over spectacle lens prescription accuracy between the optical laboratory and the practitioner is to be expected, especially with a progressive addition lens (PAL). The PAL continues to evolve in design, yet retains some unique, inherent problems. The purpose of this study was to determine if the source of these problems results from the lens design, laboratory error, or verification procedures. METHODS: Six Varilux Comfort Orma Supra progressive addition lenses were surfaced to plano distance power, and then verified using manual and automatic lensmeters. The same lenses were then re surfaced for various sphere and cylinder powers, and verified in the same manner. The results of each trial were spherocylindrically averaged, and compared to the desired result. The lenses were also verified by several incorrect methods to investigate positional tolerance during verification and possible verification errors. RESULTS: The averages for the lenses surfaced to the "prescribed" power were not far from the expected power by all measurement techniques when measured at the central DRP location. Positional errors of as little as 2 mm from the DRP center, however, can lead to erroneous results. CONCLUSION: Unless correct verification procedures are carefully followed, progressive lenses can be mistakenly identified as being in error when they are not. PMID- 12645852 TI - Promoting healthy eyes healthy people during save your vision month. PMID- 12645853 TI - Terminating the optometrist/patient relationship. PMID- 12645854 TI - Dipping into the HIPAA question box again. PMID- 12645855 TI - Pleural mesothelial cells modulate polymorphonuclear leukocyte apoptosis in empyema. AB - In bacterial empyema, the recruited polymorphonuclear leukocytes (PMN) represent important phagocytic cells involved in antibacterial defense. In this study we demonstrate that pleural fluids (PF) obtained from patients with empyema (EMP) contains significantly higher levels of granulocyte colony stimulating factor (GM CSF), and PMN incubated in empyema (EMP) pleural fluid (PF) showed significantly less apoptosis than congestive heart failure (CHF) PF. Staphylococcus aureus stimulated PMC released significantly (P < 0.001) higher levels of GM-CSF than resting PMC. Staphylococcus aureus-stimulated PMC (SPMC)-CM significantly (P < 0.001) inhibited PMN apoptosis. In SPMC-CM-incubated PMN the antiapoptotic gene Bcl-xL mRNA and protein expression was up-regulated; Bak mRNA and protein expression was down-regulated compared to control PMN. The active caspases activity significantly decreased. When SPMC-CM and EMP PF were immunodepleted with GM-CSF antibody, PMN apoptosis was significantly higher. The delay in apoptosis of PMN is in part attributable to the release of cytokine GM-CSF by activated PMC. These findings suggest that S. aureus-activated PMC extend PMN life span by modulating Bcl-xL and Bak gene expression and active caspases activity during acute inflammation and empyema. PMID- 12645856 TI - Molecular basis of rifampicin-induced inhibition of anti-CD95-induced apoptosis of peripheral blood T lymphocytes: the role of CD95 ligand and FLIPs. AB - Rifampicin modulates immune response; however, mechanisms by which it exerts these effects are incompletely understood. Recently, rifampicin has been shown to bind to and activate glucocorticoid receptors. Because of the evidence for a role of glucocorticoids in lymphocyte apoptosis, we hypothesized that rifampicin may exert its influence on the immune system by regulating apoptosis. Therefore, we examined the effect of rifampicin on signaling pathway of anti-CD95-induced apoptosis in peripheral blood lymphocytes. Rifampicin, in a concentration dependent manner, inhibited anti-CD95-induced apoptosis in both CD4+ and CD8+ T cells, which was associated with the inhibition of activation of both caspase-3 and caspase-8. In addition, rifampicin down-regulated the expression of CD95L and Bax. The inhibitory effects of rifampicin on apoptosis and caspase activation as well as its effect on the expression of CD95L and FLIPs were reversed by RU486, an antagonist of glucocorticoid receptor. These data suggest that rifampicin inhibits anti-CD95-mediated apoptosis in lymphocytes by modulating the expression of certain proteins that regulate apoptosis, at least in part, via glucocorticoid receptors. PMID- 12645857 TI - Food restriction and fish oil suppress atherogenic risk factors in lupus-prone (NZB x NZW) F1 mice. AB - Atherosclerosis-mediated coronary artery disease is a significant cause of mortality in lupus patients. Both an activated immune system and hyperlipidemia are implicated in the pathogenesis of the atherosclerotic lesions of lupus. In this study, the increases in anticardiolipin antibodies, total cholesterol, and LDL cholesterol with age were significantly lowered by fish oil and food restriction, either alone or in combination. Food restriction also significantly decreased the elevation in anti-dsDNA antibody production seen with age in ad libitum groups. Interestingly, effects of food restriction and fish oil on both lipid profile and autoantibody production were seen from a young age. Accumulation of leukocytes in the blood vessels and deposition of IgG in the glomerular mesangium also were suppressed by food restriction. Thus, beneficial effects of fish oil and food restriction on lupus nephritis and survival could be, at least in part, due to their selective effect on atherogenic risk factors. PMID- 12645858 TI - Production of TARC and MDC by naive T cells in asthmatic patients. AB - The helper (Th)2 cell-attracting chemokines thymus and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC) are ligands for the chemokine receptor CCR4. A number of cellular sources of TARC and MDC have been identified, including not only macrophages, dendritic cells, and natural killer cells, but also bronchial epithelial cells. Recent studies report that TARC and MDC may serve as pivotal chemokines for the development of Th2-dominated experimental allergen-induced asthma. This study was designed to assess TARC and MDC production by CD4+ T cells, including naive T cells and memory/effector T cells, purified from peripheral blood mononuclear cells in patients with asthma. Asthmatic subjects included in this study had mild asthmatic symptoms, positive skin test responses to house dust mite allergen, and elevated level of Dermatophagoides farinae immunoglobulin E in the sera. CD4+ T cells--CD45RA+ CD4+ T cells--as naive T cells and CD45RO+ CD4+ T cells--as memory/effector T cells- were purified by negative selection from peripheral blood mononuclear cells obtained from asthmatic patients (n = 6) and healthy controls (n = 6). These cells and established Th1/Th2 cell lines were then cultured in the presence of both anti-CD3 and -CD28 antibodies. After 48 hr of incubation, concentrations of TARC, MDC, interleukin (IL)-4, IL-5, and interferon-gamma in the supernatants were measured by enzyme-linked immunoadsorbent assay. Reverse transcriptase polymerase chain reaction was performed to analyze mRNA expression of TARC and MDC. Our results clearly showed that TARC and MDC were produced by activated CD45RA+ CD4+ T cells rather than by activated CD45RO+ CD4+ T cells, and the levels of these chemokines in the asthmatic patients were higher than those in the healthy controls. Furthermore, these chemokines production by Th2 cell lines were greater than those by Th1 cell lines, but the level were smaller than those by naive T cells. Our studies suggest that TARC and MDC are produced by naive T cells rather than by memory/effector T cells, including Th2 cells, in asthmatic patients, and these chemokines were produced at modest levels in any T-cell populations from healthy controls. Taken together, naive T cells in asthma have a peculiar function to produce TRAC and MDC, which contribute to local migration of Th2 cells into lung and lymphoid tissues, along with a function as precursor for memory/effector T cell. This novel function of naive T cells may be implicated in the development of asthma. PMID- 12645859 TI - Expression and regulation of nuclear retinoic acid receptors in human lymphoid cells. AB - Retinoids are known to play an important role in cellular growth and differentiation and more recently in the immune response. Our laboratory has previously shown that all-trans-retinoic acid (atRA) augments immunoglobulin synthesis of cord blood mononuclear cells by enhancing the synthesis of certain cytokines. Transcriptional regulatory elements, the retinoic acid nuclear receptors (RAR), could mediate the RA-induced regulation of genes, e.g., cytokines whose products are involved in the pathways of immunoglobulin synthesis. Although much is known about RAR in various animal species and tissues, little is known about the expression of RAR and its isotypes in human lymphoid cells. In this study, we examined the RAR isotypes (RAR-alpha, RAR-beta, RAR-gamma) and their respective isoforms in T- and B-lymphoid cells using a quantitative RT-PCR assay. RAR-alpha1 and -gamma1 were both constitutively expressed and did not change with the addition of atRA to human T- and B-cell lines or adenoidal T and B lymphocytes. In contrast, RAR-beta2 was not detected. The addition of atRA to cell culture produced a marked increase in the amounts of RAR-beta2 mRNA (2.2- to 41-fold). As with the RAR-beta2 isoform, the addition of atRA increased RAR-alpha2 mRNA levels (3.4- to 17-fold), but only in EBV transformed B cells and adenoidal B lymphocytes. The RAR-beta1 and -beta3 isoforms were undetectable in lymphoid cells and not inducible with atRA. RAR gamma2 was expressed at very low levels and was not inducible with atRA. Our results suggest that the expressions of the RAR-alpha2 and -beta2 isoforms in lymphoid cells are highly controlled by atRA. Differences in the regulation of RAR isoforms by atRA in human lymphoid cells may be an important factor in the modulation of cytokine production and the augmentation in Ig synthesis by atRA. PMID- 12645860 TI - Long-term follow-up and prognosis of chronic granulomatous disease in Yugoslavia: is there a role for early bone marrow transplantation? AB - We report the long-term follow-up of 12 pediatric-aged patients with chronic granulomatous disease (CGD). The mean age at the onset of infections was 5 months with a median delay in diagnosis of 2.5 years. Bacille Calmette-Guerin lymphadenitis was the most common presenting infection (6) followed by suppurative lymphadenitis (4), liver abscess (1), or Salmonella sepsis (1). Prophylaxis with cotrimoxazole was recommended to all patients. During the mean follow-up of 10 years (range, 4-23 years) pneumonitis was the most prevalent infection (91%) followed by lymphadenitis (83%), aphtous stomatitis (58%), and liver abscesses (25%). Seven (58%) patients developed chronic lung disease due to grossly delayed diagnosis (3) or poor compliance with antimicrobial prophylaxis (4). Five (41%) patients died during the second decade of life of aspergillosis (3) or chronic lung disease (2). Probability of survival into the third decade of life was estimated to be only 19%. We argue that HLA-identical bone marrow transplantation (BMT), if possible, should be attempted at early age because of significant morbidity and mortality in adolescence. BMT also should be considered in patients who suffer severe infections despite antimicrobial prophylaxis or patients with evidence of chronic lung disease. Possibility of elective BMT from unrelated donors remains to be carefully evaluated. PMID- 12645861 TI - Pilot study of an HLA-A2 peptide vaccine using flt3 ligand as a systemic vaccine adjuvant. AB - A pilot vaccine study was conducted to test the safety and immunological efficacy of four monthly immunizations of an MHC class I peptide vaccine, the E75 HLA-A2 epitope from HER-2/neu, using flt3 ligand as a systemic vaccine adjuvant. Twenty HLA-A2-expressing subjects with advanced stage prostate cancer were randomly assigned to one of four immunization or treatment schedules: (a) Flt3 ligand (20 microg/kg per day) administered subcutaneously daily for 14 days on a 28-day cycle, monthly for four months; (b) flt3 ligand course as above with the E75 peptide vaccine administered on day 7 of each flt3 ligand cycle; (c) flt3 ligand course as above with the E75 peptide vaccine administered on day 14 of each flt3 ligand cycle; or (d) E75 peptide admixed with granulocyte-macrophage colony stimulating factor and administered intradermally once every 28 days, as has previously been reported. The primary endpoints of the study were the determination of safety and immunological efficacy in generating E75-specific T cells as determined by peptide-specific interferon-gamma ELIspot. Adverse events included one grade 3 skin reaction and the development of grade 2 autoimmune hypothyroidism in two subjects with preexisting subclinical autoimmune hypothyroidism. Dendritic cells were markedly increased in the peripheral blood of subjects receiving flt3 ligand with each repetitive cycle, but augmentation of antigen-presenting cells within the dermis was not observed. Apart from a single subject, no significant peptide-specific T-cell responses were detected by ELIspot, whereas delayed-type hypersensitivity responses were detectable in control subjects and in subjects receiving peptide vaccine early in the course of flt3 ligand administration. The absence of robust peripheral immune responses in the current study may be attributable to the small numbers of subjects or differences in the subject population. In addition, the inability of fit3 ligand to augment the number of peripheral skin antigen-presenting cells may have contributed to the absence of robust peptide-specific immunity detectable in the peripheral blood of immunized subjects treated with flt3 ligand. PMID- 12645862 TI - A randomized double-blind study to compare the effectiveness of three different doses of lignocaine used in paracervical block during oocyte retrieval. AB - PURPOSE: To compare the effectiveness of three different doses of lignocaine used in paracervical block (PCB) during transvaginal ultrasound-guided oocyte retrieval (TUGOR) METHODS: In this double-blind study, 153 patients undergoing TUGOR in their first in vitro fertilization cycle were randomized to receive 50, 100, and 150 mg of lignocaine in PCB. Pain levels were measured by a 100-mm linear visual analogue scale (0 = none to 100 = intolerable). RESULTS: No differences were seen in the demographic data, the ovarian responses, the duration of TUGOR, and the number of follicles punctured. Vaginal and abdominal pain levels during TUGOR and 4 h after TUGOR were not significantly different among the three groups The median vaginal and abdominal pain levels during the retrieval were 22.0-24.0 and 30.0-32.0 respectively. CONCLUSION: The use of 50 mg of lignocaine is recommended in PCB because of the lack of improvement in pain relief on higher doses and potential dose-related risks. PMID- 12645865 TI - Confronting the hidden face of progesterone during the follicular phase. AB - Sporadic cases of late follicular phase progesterone elevation have been described in ART cycles despite routine blockage of endogenous gonadotropins with GnRH agonists, the mechanism of which has been eluding our scrutiny. In this issue of JARG, Levran et al. report the first cases of adult-onset mild form of congenital adrenal hyperplasia (CAH) linked to a deficit in 17-hydorixyloase (17 OH ase). Contrary to CAH due to deficit 21-OH ase, these not only affect the adrenals but also the FSH driven production of E2 and androgen by the ovary thereby leading to progesterone "accumulation" during the follicular phase. Levran's findings offer new hypotheses for explaining the cases late follicular progesterone elevation particularly, when encountered in COH cycles associated with poor E2 elevation. And more importantly, Levran's findings offer new possible options for minimizing their consequences on ART outcome. PMID- 12645864 TI - Familial partial 17,20-desmolase and 17alpha-hydroxylase deficiency presenting as infertility. AB - PURPOSE: Females with 17alpha-hydroxylase/17,20-desmolase deficiency normally present with amenorrhea, sexual infantilism, hypertension, and hypokalemia. We report on a new clinical presentation of this combined enzymatic defect. METHODS: Four Jewish women from two unrelated families presented with primary infertility. All patients exhibited a normal phenotype, blood pressure, and serum potassium levels with abnormally high follicular phase serum progesterone and low E2 levels. In order to characterize the underlying defect, the following steps were undertaken: 1) ovarian suppression by GnRH agonist, 2) adrenal suppression by dexamethasone, 3) ovarian stimulation by gonadotropins, 4) adrenal stimulation by ACTH, 5) hormonal assessment of follicular fluid aspirates, and 6) assessment of in vitro E2 production by luteinized granulosa cells. RESULTS: The clinical characteristics and endocrine testing results support the diagnosis of a partial deficiency in 17alpha-hydroxylase/17,20-desmolase activities, shared by the adrenal gland and the ovaries CONCLUSIONS: Female infertility can be the first and sole clinical manifestation of this enzymatic defect. Its exact nature and prevalence remain to be determined. PMID- 12645863 TI - Reduced expression of alphavbeta3 integrin in the endometrium of unexplained infertility patients with recurrent IVF-ET failures: improvement by danazol treatment. AB - PURPOSE: To determine whether there is any association between the expression of endometrial integrin alphavbeta3 and repeated IVF-ET failure and to examine the effect of danazol treatment on alphavbeta3 expression. METHODS: This prospective study was performed using a semiquantitative immunohistochemical analysis on the staining intensity of alphavbeta3 in the mid-secretory endometria derived from 10 fertile women and 57 infertile patients with a history of repeated IVF-ET failures. Nine patients randomly selected from these 22 patients with unexplained infertility were then treated with oral danazol administration for 12 weeks and reexamined at the first mid-secretory phase after the danazol treatment. RESULT(S): The levels of endometrial alphavbeta3 expression were lower in 22 patients with unexplained infertility than in the fertile control and 35 patients with explained infertility. The 9 patients treated with danazol showed a significant increase in the alphavbeta3 staining. CONCLUSION(S): The significantly decreased expression of endometrial integrin alphavbeta3 suggested that functional, but not morphological, endometrial defect may be one of the causes for the patients with unexplained infertility. Danazol may have a therapeutic potential in improving endometrial function together with up regulation of alphavbeta3. PMID- 12645866 TI - Different intervals between vasectomy and sperm retrieval interfere in the reproductive capacity from vasectomized men. AB - PURPOSE: To evaluate the relationship between the postvasectomy period and sperm reproductive capacity after ICSI. METHODS: Seventy-seven ICSI cycles with percutaneous epididymal sperm aspiration (PESA) were reviewed. Patients were divided into 4 groups according to the interval after vasectomy: 0 - 5 years (G1); 6 - 8 years (G2); 9 - 14 years (G3), and > 15 years (G4). RESULTS: Clinical and ongoing pregnancy rates did not correlate significantly with the time period of vasectomy until 14 years. Although the higher implantation rate observed in G1, no significant differences were noted among Groups 1-3. The miscarriage rates increased from G1 to G4, reaching a statistical significance among G1, G2, and G3 compared with G4. When groups were also divided according to the maternal age, the same results were obtained. CONCLUSIONS: The interval between the vasectomy and the sperm retrieval procedure has no effect on the outcome until the interval of 14 years. PMID- 12645867 TI - Can Westgard Quality Control Rules determine the suitability of frozen sperm pellets as a control material for computer assisted semen analyzers? AB - PURPOSE: To evaluate the drop-to-drop and pellet-to-pellet repeatability and stability of frozen sperm pellets. METHODS: Ten pellets were thawed per batch (low and normal concentration) and evaluated by two investigators to establish a quality control chart. Then low and normal concentration pellets were thawed and evaluated daily for 10 days by both investigators. The values for both investigators were averaged and plotted on the chart. RESULTS: The low sperm concentration specimen had a systematic error while the normal sperm concentration specimen had a random error as well as a systematic error. The low sperm concentration specimen violated the warning rule for motility whereas the normal concentration violated the warning rule, the random error rule, and the systematic error rule when applied to motility. CONCLUSIONS: Frozen sperm pellets are not acceptable as a daily-use quality control material for semen analysis when using a computer assisted semen analyzer. PMID- 12645868 TI - A new approach for screening for Y microdeletions: capillary electrophoresis combined with fluorescent multiplex PCR. AB - PURPOSE: To apply capillary electrophoresis for rapid screening for Y microdeletions METHODS: A set of 25 specific sequence tagged sites that cover the azoospermia factor a, b, and c regions of the Y chromosome was amplified in 5 fluorescent multiplex sets each including 5 primer pairs. One of the primers of each pair was labeled with a fluorescent tag attached to the 5'-end. After PCR amplification, analysis of the obtained PCR products was performed using capillary electrophoresis (ABI Prism 3100 Genetic Analyzer). The method was employed to determine Y microdeletions in azoospermic (n = 49) and severe oligozoospermic (n = 149) men. RESULTS: The number of PCR cycle (from 45 to 30) and the amount of DNA template (20-fold) used in fluorescent multiplex PCR were reduced because of the high sensitivity of capillary electrophoresis. Approximately 1000 multiplex PCR sets from 198 patients were analyzed simultaneously within 50 h. Y microdeletions were found in 3 out of the 198 azoospermic or severe oligozoospermic men. CONCLUSIONS: Application of capillary electrophoresis for detection of PCR products provides a semiautomated, high throughput method for rapid screening for microdeletions on the Y chromosome. PMID- 12645869 TI - Genetic variation in the midcontinental population of sandhill cranes, Grus canadensis. AB - Three subspecies of sandhill crane (Grus canadensis) are recognized in the Midcontinental population, the lesser (Grus c. canadensis), Canadian (G. c. rowani), and greater (G. c. tabida). Blood samples collected on the population's primary spring staging area in Nebraska, U.S.A., were used to resolve the genetic relationship among these subspecies. Phylogenetic analysis of 27 G. canadensis, by DNA sequencing of a 675 bp region of the mtDNA, supports the subspecies designations of G. c. canadensis and G. c. tabida. G. c. rowani individuals were intermediate with each of the other two subspecies. Genetic divergence ranged from 6.5 to 14.5% between G. c. canadensis and G. c. tabida, 0.5 to 6.6% within G. c. canadensis, and 0.1 to 6.0% within G. c. tabida. Sufficient DNA for analysis was obtained from shed feathers indicating a source of genetic material that does not require the capture or sacrifice of the birds. Other genetic markers and methods, including satellite telemetry, are required for obtaining detailed information on crane distributions as needed to establish effective management units for the MCP. PMID- 12645870 TI - Cloning, expression, and genomic structure of a novel human Rap2 interacting gene (RPIP9). AB - During a large-scale screen of a human fetal brain cDNA library, a full-length cDNA encoding a novel Rap2 interacting protein was isolated and sequenced. The cDNA is 3397bp long and has a predicted open reading frame encoding a protein of 329 aa. The predicted protein shows high homology to mouse and human RPIP8, and has a RUN domain near its C-terminus. The gene was mapped to human chromosome 7q21-7q22 and has 9 exons and 8 introns. The expression pattern was also detected by cycle-limited reverse polymerase chain reaction (RT-PCR). PMID- 12645871 TI - Twelve single nucleotide polymorphisms on chromosome 19q13.2-13.3: linkage disequilibria and associations with basal cell carcinoma in Danish psoriatic patients. AB - The genetic susceptibility to basal cell carcinoma (BCC) among Danish psoriatic patients was investigated in association studies with 12 single nucleotide polymorphisms on chromosome 19q13.2-3. The results show a significant association between BCC and the A-allele of a polymorphism in ERCCI exon4 (Odds ratio 12;95% Confidence Interval 1.17-124; p(chi2, two-side) = 0.019) and to a lesser extent with XPD exon6 (p = 0.06). This is in accordance with recent studies of a different group of BCC cases (Rockenbauer et al. (in press) Carcinogenesis; Yin et al. (manuscript submitted for publication). Cancer Epidemiol. Biomarkers Prev), which places two highly influential markers between these two genes. The analysis also confirmed that considerable linkage disequilibrium exists between SNPs both within genes and between genes in this region. The combined studies suggest that genetic variation in nucleotide excision repair is of importance for the development of BCC. PMID- 12645872 TI - High mobility group-like proteins of the insect Plodia interpunctella. AB - Nuclei from Plodia interpunctella larvae contain four major proteins, which are extracted by 5% perchloric acid and 0.35 M NaCl. The proteins have been designated PL1, PL2, PL3, and PL4. The amino acid analyses of these proteins show that they have high proportions of acidic and basic amino acid residues, a property characteristic of the high mobility group (HMG) proteins isolated from vertebrate tissues. Immunological characterication of these proteins clearly shows that PL1, PL2, and PL4 are more closely related to HMG1 dipteran proteins, while PL3 is more closely related to HMG1 dipteran proteins. The possible relatedness of these proteins to HMG proteins is discussed. PMID- 12645873 TI - Allozyme variation and population differentiation of the Aconitum delavayi complex (Ranunculaceae) in the Hengduan Mountains of China. AB - The Aconitum delavayi complex is a group of four climbing species with trisectleaves occurring in the Hengduan Mountains. The species of this complex are highly localized on very narrow regions with quite small population sizes. Because of rapid environmental changes recently in the Hengduan Mountains, this complex shows complicated morphological variability which makes it difficult to delimit species. In the present study, 10 enzyme systems coding for 14 putative loci were employed to detect the interspecific and intraspecific genetic variation of the complex. In addition to low genetic diversity within all eight populations surveyed, the results indicate that A. episcopale is a distinct species because of high genetic identities among its three populations. Very low genetic divergence among populations of A. stapfianum and A. delavayi suggests that the two species should be treated as a single one. PMID- 12645874 TI - An improved COII primer specific for Lepidoptera. PMID- 12645875 TI - Pathways to a robust immune response in the elderly. AB - Circumstantial evidence suggests that infectious disease is the major cause of morbidity and mortality in the elderly, and immune-system dysfunction may contribute to this finding. Because innate and humoral immunity seem to be relatively unaffected by aging and because the T-cell compartment shows marked age-associated alterations, this article focuses on the association between T cells and aging. Longitudinal studies suggest that immune parameters, which predominantly are related to T cells, can be clustered to yield an IRP that is predictive of mortality in the elderly. Determining the IRP also may be helpful in younger individuals, particularly those under chronic antigenic stress (eg, patients with cancer or chronic infections) who experience premature aging of the immune system. Some changes in T cells can be modeled in clonal cultures in vitro to discover new biomarkers of immune aging. These biomarkers, which need to be validated in vivo, could be used to refine IRP. Interventions to selectively target changes that are identified as part of IRP may improve the health and quality of life of the elderly, reduce healthcare costs, and avoid potential unwanted side effects of global intervention approaches, such as triggering or exacerbating autoimmunity and inflammation. PMID- 12645876 TI - Age-related inflammatory cytokines and disease. AB - Aging is associated with chronic low-grade increases in circulating levels of inflammatory markers. A wide range of environmental factors, including smoking, infections, and obesity, genetic factors, and the declining function of sex hormones may contribute to systemic low-grade inflammatory activity in older individuals. Age-associated disease may exacerbate this phenomenon. The multifunctional cytokines TNF-alpha and IL-6 have been associated with morbidity and mortality in the elderly. Evidence supports the direct role of TNF-alpha in the pathogeneses of atherosclerosis, type 2 DM, and AD in older individuals. Age related increases in systemic levels of TNF-alpha could provide a unifying basis for these disorders. Furthermore, TNF-alpha induces a catabolic state that causes frailty. Circulating levels of IL-6 seem to be a strong risk factor for frailty in the elderly, which could reflect its association with increased production of TNF-alpha. IL-6 also may be a risk factor for thromboembolic complications. In healthy, elderly populations, high circulating levels of TNF-alpha and IL-6 predict mortality, independent of comorbidity, indicating that TNF-alpha and IL-6 cause morbidity and mortality. In cohorts of frail, older individuals, TNF-alpha and IL-6 also act as disease markers. Circulating levels of TNF-alpha seem to be the best predictor of mortality in frail, elderly populations with a high mortality rate, whereas IL-6 seems to be the strongest risk marker in healthy, elderly populations. This finding could reflect that in relatively healthy old populations the increase in circulating levels of IL-6 represent a systemic response to local proinflammatory activities; however, when age-related inflammatory diseases progress, levels of TNF-alpha increase in the circulation and become gradually a stronger risk marker than IL-6. In conclusion low-grade elevations in levels of circulating cytokines are strong independent risk factors of morbidity and mortality in the elderly, and lifestyle factors and comorbidities may modulate these levels. Exercise and dietary interventions may be possible strategies to decrease inflammatory activity and improve the health status of the elderly. PMID- 12645877 TI - Problems and solutions to the development of vaccines in the elderly. AB - Scientists involved in vaccine research and development face the challenge of protecting the ever-increasing elderly population from a broad spectrum of infectious diseases. The optimal vaccine-induced immune response to confer protection is undefined for many pathogens, and the field of vaccine research is undergoing a gradual shift from the original focus on humoral immunity to a focus that incorporates cellular and innate immune components. The age-related changes in various aspects of immune function, including an increase in a population of T cells that shows signs of replicative senescence, underscore the need to enhance research aimed at designing vaccines to meet the unique requirements of the elderly population. PMID- 12645878 TI - Infectious disease risk in the elderly. AB - Older persons suffer excessively from infectious diseases such as pneumonia and urinary tract infections. This article discusses some of the reasons for this additional morbidity and mortality, including the anatomical and physiological changes with aging, impairment of immune function, presence of co-morbid diseases, and delays in diagnosis and initiation of therapy. PMID- 12645879 TI - Origin of late-onset autoimmune disease. AB - Autoimmune disease in the elderly is hypothesized to be caused by an imbalance in T-cell expansion and deletion after an encounter with self-antigens. A decrease in thymic output leads to a decreased pool of naive T cells in the periphery and to increased oligoclonal expansion of T cells. This expansion may be caused by stimulation with autoantigens that drive high-affinity interactions with self antigens. Accumulation of presenescent, apoptosis-resistant, and proinflammatory T cells results in the growth of these autoreactive T cells. A decreased T-cell activation response that occurs with age leads to several defects that diminish the immune response. PMID- 12645881 TI - Immunologic and inflammatory mediators and cognitive decline in Alzheimer's disease. AB - Local up-regulation of immune mediators is implicated in the development of AD; however, the systemic cell-mediated immune response in patients with AD may be impaired. Progression of AD from moderate to severe stages is accompanied by a progressive decline in cognitive function, independence, and behavioral and motor impairments and by the occurrence of recurrent infections, such as bronchitis and pneumonia. Several factors, including malnutrition, may be implicated in the reduced ability to mount an adequate immune response against invading pathogens in advanced stages of AD. The immune system has an important role in the development, progression, and potential treatment of AD and is involved in determining the subsequent sequelae and morbidity outcomes in patients with advanced-stage AD. A better understanding of the immune mediators that are involved in the local and systemic immune response in AD may assist in mediating the development of AD, halting its progression, and treating patients with anti inflammatory drugs and immunization strategies. PMID- 12645880 TI - Aging, immunity, and tumor susceptibility. AB - The study of aging, susceptibility to tumors, and immunity is at an initial stage, and numerous excellent questions challenge immunologists and tumor biologists. Because unpredicted factors contribute to tumor susceptbility in the human population, the authors feel that the development of an animal model is essential to dissect these complicated issues. PMID- 12645883 TI - Future challenges in analysis and treatment of human immune senescence. AB - Immune functions decline with age; immunorestorative approaches have been explored in laboratory animals and humans with definite, but limited success. In the clinical setting, the age-associated immune deficiency (immunesenescence) is compounded by the presence of comorbidities that are associated with a functional decline in immunity. Thus, any successful immune reconstitution strategy for humans must involve treatment of the underlying diseases. Furthermore, general health measures such as nutrition and exercises may have powerful effects on restoring immunity, to the extent that malnutrition and a sedentary lifestyle have contributed to existing immune deficiency. More selective approaches, such as with specific cytokines or organ transplantation are of experimental interest but are quite distant from achieving clinical applicability as interventions to restore immunity in the frail elderly. PMID- 12645882 TI - Atherosclerosis as a paradigmatic disease of the elderly: role of the immune system. AB - When a new hypothesis about the etiology and pathogenesis of a disease is developed, there is always the danger that it will be presented as the only acceptable explanation for the occurrence of a given pathologic condition. In view of the well-proven multifactoral pathogenesis of atherosclerosis, we would like to emphasize that we are not postulating that immunity to HSP60 is the only cause of atherogenesis, especially in the later stages where there are clinically apparent sequelae, such as myocardial infarction, stroke, and other atherosclerosis-dependent symptoms. In this article, we summarized some of the experimental and clinical data that we and others have collected in support of the concept that atherosclerosis is a good example of pleotropic antagonism, and postulated that age-dependent diseases are the price we pay for genetic traits established by natural selection to assure maximum survival until the age of reproduction, the effects of which may, however, become deleterious later in life. In the present case, the cost we pay for protective immunity to microbial and altered autologous HSP60 is the risk of cross-reactivity with HSP60 expressed by arterial endothelial cells that are subjected to stress factors already known as classical atherosclerosis risk factors. We showed that the first inflammatory stage of atherosclerosis starts early in life, long before it becomes clinically apparent. More severe lesions that lead to atherosclerosis-dependent organ specific or systemic symptoms will only occur if classical atherosclerosis risk factors, especially those involving the cholesterol metabolism, remain present. PMID- 12645884 TI - Glycosylation, who cares? PMID- 12645885 TI - Distinguishing between phosphorylated and nonphosphorylated peptides with ion mobility-mass spectrometry. AB - Mass spectrometry has become an indispensable tool in identifying post translationally modified proteins, but multiple peptide mass-mapping/peptide sequencing experiments are required to answer questions involving the site and type of modification present. Here, we apply ion mobility-mass spectrometry (IM MS), a high-throughput analysis method having high selectivity and sensitivity, to the challenge of identifying phosphorylated peptides. Ion mobility separation is based on the collision cross-section of the ion. Phosphorylation can result in a conformational change in gas-phase peptide ions, which can be detected by IM. To demonstrate this point, a peptide mixture containing a variety of peptide sequences is examined with IM-MS and molecular dynamics calculations. During the course of these studies, two classes of phosphopeptide were identified: (i) phosphorylated peptide ions that have conformers that differ from the nonphosphorylated ion and (ii) phosphorylated peptide ions that have conformations that are very similar to the nonphosphorylated peptide. The utility of IM-MS peptide mass mapping for identifying both types of phosphorylated peptides is discussed. PMID- 12645886 TI - Structural and functional properties of Yersinia pestis Caf1 capsular antigen and their possible role in fulminant development of primary pneumonic plague. AB - Yersinia pestis capsular antigen Caf1 is shown to be a beta-structural protein that in polymeric form possesses very high conformational stability. Different approaches show that a dimer is the minimal cooperative block of Caf1 adhesin. Caf1 dimer interacts effectively with IL-1 receptors of human macrophage and epithelial cells. The specificity of such interaction is confirmed by the inhibition of IL-1alpha binding by Caf1. The Caf1 role in pneumonic plague pathogenesis is discussed. PMID- 12645887 TI - Quantitative profiling of proteins in complex mixtures using liquid chromatography and mass spectrometry. AB - The objective of this study was to determine if liquid chromatography mass spectrometry (LC/MS) data of tryptic digests of proteins can be used for quantitation. In theory, the peak area of peptides should correlate to their concentration; hence, the peak areas of peptides from one protein should correlate to the concentration of that particular protein. To evaluate this hypothesis, different amounts of tryptic digests of myoglobin were analyzed by LC/MS in a wide range between 10 fmol and 100 pmol. The results show that the peak areas from liquid chromatography mass spectrometry correlate linearly to the concentration of the protein (r2 = 0.991). The method was further evaluated by adding two different concentrations of horse myoglobin to human serum. The results confirm that the quantitation method can also be used for quantitative profiling of proteins in complex mixtures such as human sera. Expected and calculated protein ratios differ by no more than 16%. We describe a new method combining protein identification with accurate profiling of individual proteins. This approach should provide a widely applicable means to compare global protein expression in biological samples. PMID- 12645888 TI - Defining absolute confidence limits in the identification of Caulobacter proteins by peptide mass mapping. AB - A derivatization reaction, guanidination, was recently reported that increases MALDI-TOF MS sensitivity toward lysine-terminated peptides. Its application conveys sequence information that can be used as a parameter in peptide mass mapping database searches. This paper presents a systematic study of the impact of guanidination on proteomic analysis of an entire bacterial organelle. Sixty two 2-D gel isolated proteins from Caulobacter crescentus stalks were studied. A novel computer algorithm, Prodigies, was developed to analyze the data. Absolute confidence limits associated with protein assignments were established using Monte Carlo simulations of database searches. The advantages of guanidination are illustrated using both experimental and theoretical data. PMID- 12645889 TI - Array-based multiplexed screening and quantitation of human cytokines and chemokines. AB - HydroGel-coated slide is a porous substrate based on a polymer matrix that provides a three-dimensional hydrophilic environment similar to free solution suitable for biomolecular interactions. This substrate has been used to develop fluorescence-based multiplexed cytokine immunoassays. Forty-three monoclonal antibodies (mAb) of cytokines and chemokines were printed at a volume of 350 pL per spot using a Packard BioChip Arrayer. For each probe, four replicates were printed at a pitch of 500 microm in the layout of a 13 x 16 pattern on a 12 x 12 mm2 HydroGel pad. Cytokines and chemokines that are captured by the arrayed mAbs are detected by using another biotinylated mAb, following by the addition of a Texas Red-conjugated streptavidin. The fluorescent images of arrays were recorded using a Packard ScanArray 5000 confocal slide scanner and quantitated using Packard QuantArray software. Experiments demonstrated that 43 cytokines and chemokines could be simultaneously screened and quantitated in conditioned culture media, cell lysates, and human plasma. Using this chip, we have examined cytokine expression in breast cancer cells and identified the chemokines associated with human cervical cancers. PMID- 12645890 TI - Quantitative analysis of the yeast proteome by incorporation of isotopically labeled leucine. AB - Quantitative comparison of protein expression levels in 2D gels is complicated by the variables associated with protein separation and mass spectrometric responses. Metabolic labeling allows cells from different experiments to be mixed prior to analysis. This approach has been reported for prokaryotic cells. Here, we demonstrate that metabolic labeling can also be successfully applied to the eukaryote Saccharormyces cerevisiae. Yeast leucine auxotrophs grown on synthetic complete media containing natural abundance Leu or D10-Leu were mixed prior to 2D gel separation and MALDI analysis of the digested proteins. D10-Leu labeling provided an effective internal calibrant for peptide MS analysis, and the number of Leu residues yielded an additional parameter for peptide identification at low mass resolution (1000). Metabolic incorporation of D10-Leu into yeast proteins was found to be quantitative since the intensities of the peptide peaks corresponded to those expected on the basis of the percent label in the media. Thus, D10-Leu labeling should provide reliable data for comparing proteomes both quantitatively and qualitatively from wild-type and nonessential-gene-null-mutant strains of S. cerevisiae. Given the central role played by yeast in our understanding of eukaryotic gene and protein expression, it is anticipated that the quantitative expressional proteomic method outlined here will have widespread applications. PMID- 12645892 TI - Rapid analysis of tryptically digested cerebrospinal fluid using capillary electrophoresis-electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry. AB - Mass spectrometry has in recent years been established as the method of choice for protein identification and characterization in proteomics. Capillary electrophoresis (CE) is a fast and efficient method for the separation of peptides and proteins. The on-line combination of CE with Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS) has been shown to be a powerful tool in the analysis of complex mixtures of proteins. This paper presents the first results from a proteomic analysis of human cerebrospinal fluid proteins by tryptic digestion and CE-FTICR-MS, where 30 proteins could be identified on a 95% confidence level with mass measurement errors less than 5 ppm. PMID- 12645891 TI - Enrichment of integral membrane proteins for proteomic analysis using liquid chromatography-tandem mass spectrometry. AB - An increasing number of proteomic strategies rely on liquid chromatography-tandem mass spectrometry (LC-MS/MS) to detect and identify constituent peptides of enzymatically digested proteins obtained from various organisms and cell types. However, sample preparation methods for isolating membrane proteins typically involve the use of detergents and chaotropes that often interfere with chromatographic separation and/or electrospray ionization. To address this problem, a sample preparation method combining carbonate extraction, surfactant free organic solvent-assisted solubilization, and proteolysis was developed and demonstrated to target the membrane subproteome of Deinococcus radiodurans. Out of 503 proteins identified, 135 were recognized as hydrophobic on the basis of their calculated hydropathy values (GRAVY index), corresponding to coverage of 15% of the predicted hydrophobic proteome. Using the PSORT algorithm, 53 of the proteins identified were classified as integral outer membrane proteins and 215 were classified as integral cytoplasmic membrane proteins. All identified integral cytoplasmic membrane proteins had from 1 to 16 mapped transmembrane domains (TMDs), and 65% of those containing four or more mapped TMDs were identified by at least one hydrophobic membrane spanning peptide. The extensive coverage of the membrane subproteome (24%) by identification of highly hydrophobic proteins containing multiple TMDs validates the efficacy of the described sample preparation technique to isolate and solubilize hydrophobic integral membrane proteins from complex protein mixtures. PMID- 12645893 TI - Insight into residues critical for antithrombin function from analysis of an expanded database of sequences that includes frog, turtle, and ostrich antithrombins. AB - Complete sequences were determined for frog, turtle, and ostrich antithrombins. Protein sequence comparisons with the other 10 known antithrombin sequences and with sequences of other serpins have provided striking evidence for the conservation of the heparin activation mechanism and new insight into those residues important for heparin binding, for heparin activation, and for reactive center loop function, as well as an indication of which glycosylation sites might be needed for function. Importantly, an understanding of, as yet, poorly understood antithrombin-protein interactions will be greatly aided by this expanded database and comparative analysis. PMID- 12645894 TI - ProteinChip technology: a new and facile method for the identification and measurement of high-density lipoproteins apoA-I and apoA-II and their glycosylated products in patients with diabetes and cardiovascular disease. AB - This paper describes a ProteinChip technology for the identification and quantification of apolipoprotein profiles in crude biological samples. Expression levels of apoA-I and apoA-II and their glycosylated products were accomplished using single 1 microL plasma samples. In the present studies, strong anionic and weak cationic exchanger ProteinChips (SAX2 and WCX2 chip surfaces) were tested, and the WCX2 chip was found to be selective for specific apolipoproteins. Using the WCX2 chip and analysis via surface-enhanced laser desorption ionization mass spectrometry (SELDI-MS), apoA-I and apoA-II were separated as sharp peaks at 28 and 17 kD and did not overlap with other serum protein peaks. Since these assays can be completed on a large number of clinical samples in approximately 1 h, further development of this technique will facilitate both epidemiological studies and therapeutic trials in assessing the role of the apolipoproteins and their glycosylated products in atherosclerosis. PMID- 12645895 TI - The importance of databases. PMID- 12645896 TI - Rapid detection of protein tyrosine kinase activity in recombinant yeast expressing a universal substrate. AB - Yeast two-hybrid systems are powerful proteomics tools for the discovery of protein-protein interactions. However, these systems are typically unable to detect interactions dependent on post-translational modifications such as tyrosine phosphorylation. We report a novel yeast tribrid system that expresses a potentially universal protein tyrosine kinase (PTK) substrate to detect diverse PTKs. Validation with the oncogenic kinases v-Abl and v-Src, which exhibit divergent substrate specificities, demonstrated significant potential for cloning PTKs en masse from cDNA libraries. PMID- 12645897 TI - Code developments to improve the efficiency of automated MS/MS spectra interpretation. AB - We report the results of our work to facilitate protein identification using tandem mass spectra and protein sequence databases. We describe a parallel version of SEQUEST (SEQUEST-PVM) that is tolerant toward arithmetic exceptions. The changes we report effectively separate search processes on slave nodes from each other. Therefore, if one of the slave nodes drops out of the cluster due to an error, the rest of the cluster will carry the search process to the end. SEQUEST has been widely used for protein identifications. The modifications made to the code improve its stability and effectiveness in a high-throughput production environment. We evaluate the overhead associated with the parallelization of SEQUEST. A prior version of software to preprocess LC/MS/MS data attempted to differentiate the charge states of ions. Singly charged ions can be accurately identified, but the software was unable to reliably differentiate tandem mass spectra of +2 and +3 charge states. We have designed and implemented a computational approach to narrow charge states of precursor ions from nominal resolution ion-trap tandem mass spectra. The preprocessing code, 2to3, determines the charge state of the precursor ion using its mass-to charge ratio (m/z) and fragment ions contained in the tandem mass spectrum. For each possible charge state the program calculates the expected fragment ions that account for precursor ion m/z values. If any one of the numbers is less than an empirically determined threshold value then the spectrum corresponding to that charge state is removed. If both numbers are higher than the threshold value then +2 and +3 copies of the spectrum are kept. We present the comparison of results from protein identification experiments with and without using 2 to 3. It is shown that by determining the charge state and eliminating poor quality spectra 2to3 decreases the number of spectral files to be searched without affecting the search results. The decrease reduces computer requirements and researcher efforts for analysis of the results. PMID- 12645898 TI - On characterization of dose variations of 2-D proteomics maps by matrix invariants. AB - We explore the characterization of 2-D electrophoresis proteomics maps by certain structural invariants derived from matrixes constructed by considering for all pairs of spots in a proteomics maps the shortest (Euclidean) distances and distances measured along zigzag lines connecting protein spots of the neighboring abundance. This paper is a sequel to previous papers in which we outlined the idea of characterizing 2-D proteomics maps by graph-theoretical descriptors. To illustrate the approach, we selected data of Anderson et al. (Anderson, N. L.; Esquer-Blasco, R.; Richardson, F.; Foxworthy, P.; Eacho, P. The effects of peroxisome proliferators on protein abundances in mouse liver. Toxicol. Appl. Pharmacol. 1996, 137, 75-89) on protein abundance in mouse liver under a series of dose of peroxisome proliferator LY1711883. We found strong linear correlation between the experimentally applied doses and the leading eigenvalue of a D/D-type matrix (Randic, M.; Kleiner, A. F.; DeAlba, L. M. Distance/ distance matrices. J. Chem. lnf. Comput. Sci. 1994, 34, 277-286) constructed for the experimental proteomics maps. PMID- 12645899 TI - Antibody detection in human serum using a versatile protein chip platform constructed by applying nanoscale self-assembled architectures on gold. AB - We report a novel high-throughput (HTP) protein chip platform, constructed on gold using self-assembly techniques, for conducting high quality antigen-antibody interactions. Biotinylated monolayers were used to immobilize a streptavidin surface with high packing density. This biocompatible platform was then used for detection of serum IgM antibodies. Serum samples of patients suspected to suffer from Lyme borreliosis were used to validate the protein chip platform using biotinylated peptide AAOspC8 molecules as the test probes. Various experimental parameters such as the effect of concentration of probes, targets, temperature of incubation, and their effect on the resulting signal-to-noise ratio are described in detail. Highly specific protein interaction data with a high signal-to-noise ratio were obtained with serum sample solutions as low as 1 microL/spot (1/10 diluted). PMID- 12645900 TI - Elevated HGF levels in sera from breast cancer patients detected using a protein microarray ELISA. AB - We developed an ELISA in high-density microarray format to detect hepatocyte growth factor (HGF) in human serum. The microassay can detect HGF at sub-pg/mL concentrations in sample volumes of 100 microL or less. The microassay is also quantitative and was used to detect elevated HGF levels in sera from recurrent breast cancer patients. The microarray format provides the potential for high throughput quantitation of multiple biomarkers in parallel, as demonstrated with a multiplex analysis of five biomarker proteins. PMID- 12645901 TI - Integrating 'top-down" and "bottom-up" mass spectrometric approaches for proteomic analysis of Shewanella oneidensis. AB - Here we present a comprehensive method for proteome analysis that integrates both intact protein measurement ("top-down") and proteolytic fragment characterization ("bottom-up") mass spectrometric approaches, capitalizing on the unique capabilities of each method. This integrated approach was applied in a preliminary proteomic analysis of Shewanella oneidensis, a metal-reducing microbe of potential importance to the field of bioremediation. Cellular lysates were examined directly by the "bottom-up" approach as well as fractionated via anion exchange liquid chromatography for integrated studies. A portion of each fraction was proteolytically digested, with the resulting peptides characterized by on line liquid chromatography/tandem mass spectrometry. The remaining portion of each fraction containing the intact proteins was examined by high-resolution Fourier transform mass spectrometry. This "top-down" technique provided direct measurement of the molecular masses for the intact proteins and thereby enabled confirmation of post-translational modifications, signal peptides, and gene start sites of proteins detected in the "bottom-up" experiments. A total of 868 proteins from virtually every functional class, including hypotheticals, were identified from this organism. PMID- 12645903 TI - Additive method for the prediction of protein-peptide binding affinity. Application to the MHC class I molecule HLA-A*0201. AB - A method has been developed for prediction of binding affinities between proteins and peptides. We exemplify the method through its application to binding predictions of peptides with affinity to major histocompatibility complex class I molecule HLA-A*0201. The method is named "additive" because it is based on the assumption that the binding affinity of a peptide could be presented as a sum of the contributions of the amino acids at each position and the interactions between them. The amino acid contributions and the contributions of the interactions between adjacent side chains and every second side chain were derived using a partial least squares (PLS) statistical methodology using a training set of 420 experimental IC50 values. The predictive power of the method was assessed using rigorous cross-validation and using an independent test set of 89 peptides. The mean value of the residuals between the experimental and predicted pIC50 values was 0.508 for this test set. The additive method was implemented in a program for rapid T-cell epitope search. It is universal and can be applied to any peptide-protein interaction where binding data is known. PMID- 12645902 TI - Use of MEDUSA-based data analysis and capillary HPLC-ion-trap mass spectrometry to examine complex immunoaffinity extracts of RBAp48. AB - To examine the Jurkat cell interaction partners of RbAp48, we digested entire immunoaffinity extracts with trypsin and identified potential interacting proteins using one- and two-dimensional microcapillary HPLC-ion-trap mass spectrometry. An Oracle-based automated data analysis system (MEDUSA) was used to compare quadruplicate anti-RbAp48 antibody affinity extracts with two sets of quadruplicate control extracts. The anti-RbAp48 extracts contained over 40 difference 1D gel bands. We identified all known proteins of the NuRD/Mi-2 complex including human p66. Three potential homologues of members of this complex were also found, suggesting that there may be more than one variant of this complex. Eleven proteins associated with RNA binding or pre-mRNA splicing were observed. Four other proteins, including a putative tumor suppressor, were identified, as were 18 ribosomal proteins. There was little overlap with RbAp48 interacting proteins defined by yeast two-hybrid methods. These results demonstrate the analysis of a complex immunoaffinity extract and suggest a more complex cellular role for RbAp48 than previously documented. PMID- 12645904 TI - Rapid determination of protein molecular weight by the Ferguson method and multiplexed capillary electrophoresis. AB - A method based on the use of the Ferguson method and multiplexed sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) with UV detection was demonstrated for the rapid determination of molecular weights of proteins. The method employs a capillary array where different uncoated capillaries are filled with gel buffers containing different concentrations of poly(ethylene oxide) (PEO). All data required to construct Ferguson plots and universal calibration curves for the determination of the molecular weights of diverse types of proteins are generated simultaneously in an eight-capillary array within 20 min. PMID- 12645905 TI - Capillary chromatography-coupled mass spectrometry with column switching for rapid identification of proteins from 2-dimensional electrophoresis gels. AB - An improved capillary liquid chromatography procedure, incorporating column switching in combination with mass spectrometry, is reported. The dual column system allows for rapid inject-to-inject cycle times to improve the speed of protein identification for proteomics applications. Full gradient elution of peptides from either of the two C18 columns can be achieved in less than 17 min while maintaining sufficient resolution for the peptides to be detected and fragmented by the mass spectrometer for protein identification. Importantly, the use of two columns for subsequent injections is reproducible and without carry over. The limit of detection for the system is between 25 and 50 fmol per injection. This fully automated system is capable of analyzing and identifying proteins from an entire 96-well plate in about 27 h. PMID- 12645907 TI - Results of ZD0473 in platinum-pretreated ovarian cancer: analysis according to platinum free interval. AB - Resistance to platinum-containing regimens can develop in many women with ovarian cancer and may lead to relapse in > 80% of patients. ZD0473 is a new-generation platinum agent that, in preclinical studies, shows evidence of antitumour activity and overcomes platinum-resistance mechanisms. This Phase II trial has evaluated the efficacy and tolerability of ZD0473 in second-line ovarian cancer patients. Patients received ZD0473 120 mg/m2 (1-h iv infusion, day 1 q 3-weeks); the starting dose was increased to 150 mg/m2 after a safety review. We report here on results when patients are divided into four cohorts depending upon whether they were considered platinum-resistant or -sensitive. Patients were placed into one of 3 cohorts if they were platinum resistant (relapsed/progressed < or = 26 weeks after completion of prior platinum-based chemotherapy) or cohort 4 if this period was > 26 weeks (sensitive). Ninety-four patients were recruited to the trial (59 resistant, 35 sensitive; median age 58 [range 27-75] years; 86 with performance status [PS] < or = 1). Forty-nine patients received a starting dose of 120 mg/m2, of which 15 escalated to 150 mg/m2, and 45 received a starting dose of 150 mg/m2. Overall, the median number of treatment cycles received was 3 (range 1-8). Grade 3/4 thrombocytopenia was the most common haematological adverse event occurring in 62% of patients overall. Grade 3/4 lethargy, vomiting and nausea were the most common non-haematological toxicities. No clinically significant oto-, nephro- or neurotoxicity was observed. Overall response rates for all platinum-resistant and -sensitive patients were 8.3% and 32.4%, respectively. Stable disease occurred in 17 resistant and 15 sensitive patients. PMID- 12645906 TI - ZD0473 pharmacokinetics in Japanese patients: a Phase I dose-escalation study. AB - ZD0473 is new platinum agent that was rationally designed to circumvent platinum resistance and reduce the potential for nephro-and neurotoxicity. This Phase I dose-escalating study investigated the pharmacokinetics, tolerability and efficacy of ZD0473 in Japanese patients with solid, refractory tumours. ZD0473 was administered as a 1-h intravenous infusion every 3 weeks. Nine patients received a total of 16 cycles of ZD0473 (median 1 cycle/patient), with 3 patients treated at each of 3 doses (60, 90, 120 mg/m2). The maximum plasma concentration (C(max)) and the area under the concentration-time curve to infinity (AUC(0 infinity)) increased with dose in a linear fashion for both total platinum and ZD0473 in plasma ultrafiltrate, suggesting that the pharmacokinetics of ZD0473 are linear. Haematological and non-haematological toxicities such as nausea and vomiting were mild (grade 1 or 2) and transient. No clinically significant nephro , oto- or neurotoxicity was observed. Dose-limiting toxicity (DLT) was not observed and the maximum tolerated dose (MTD) was not identified. ZD0473 treatment showed evidence of disease stabilisation in 3 patients (33%). In conclusion, ZD0473 appears to have linear pharmacokinetics, and an acceptable tolerability profile at doses up to 120 mg/m2 in Japanese patients with refractory solid malignancies. Following evaluation of the data from all the Western trials, the ZD0473 development programme changed and this Japanese trial was stopped. PMID- 12645908 TI - ZD0473 treatment in lung cancer: an overview of the clinical trial results. AB - Three open-label, non-comparative, multicentre Phase II trials have examined the efficacy and tolerability of ZD0473 as first-and second-line therapy in non-small cell lung cancer (NSCLC) patients and second-line therapy in small-cell lung cancer (SCLC) patients. Patients with second-line NSCLC or SCLC were evaluated as either platinum-sensitive or -resistant, based upon their time to relapse/progression after platinum-based therapy. First-line NSCLC patients (n = 18) received a total of 60 treatment cycles (median number per patient 2.5) whilst second-line NSCLC (n = 50) and SCLC (n = 48) patients both received a total of 127 treatment cycles (median number per patient 2.0). Grade 3/4 anaemia, neutropenia and thrombocytopenia was observed in: 38.8%, 22.2% and 27.7% of first line NSCLC patients; 12.0%, 24.0% and 50% of second-line NSCLC patients; and 10.4%, 25.0% and 47.9% of second-line SCLC patients, respectively. The most common grade 3/4 non-haematological toxicities in all three trials were lethargy and dyspnoea. No clinically significant oto-, nephro- or neurotoxicity was observed. The first-line treatment of NSCLC produced an overall response rate (OR) of 6.3%. No OR was seen after second-line treatment of NSCLC, while ORs of 15.4% and 8.3% were seen in the platinum-resistant and -sensitive second-line SCLC patients, respectively. PMID- 12645909 TI - ZD0473 combined with other chemotherapeutic agents for the treatment of solid malignancies. AB - Platinum-based combination chemotherapy regimens are the mainstay of current treatments for advanced solid malignancies. Preclinical in vitro studies have shown synergism with ZD0473 in combination with several agents, including vinorelbine and topotecan. This paper reviews the tolerability and activity observed with ZD0473 in combination with vinorelbine or topotecan, in two Phase I dose-escalating studies in patients with advanced, solid, refractory malignancies. Twenty-four patients were included in the ZD0473 plus vinorelbine trial and were treated with doses of ZD0473 60-150 mg/m2 and vinorelbine 15-25 mg/m2. In this trial, dose-limiting toxicity comprised non-haematological events and the most common grade 3/4 toxicities included neutropenia (54.2%), thrombocytopenia (29.2%) and anaemia (20.8%). Eleven patients were included in the ZD0473 plus topotecan trial and were treated with ZD0473 60-90 mg/m2 and topotecan 0.5 mg/m2/day for 3 or 5 days. In this trial, dose-limiting toxicity comprised haematological events and the most common grade 3/4 toxicities included thrombocytopenia (63.6%), neutropenia (36.4%) and anaemia (18.2%). No objective responses were observed in either trial, but disease stabilisation occurred in 29.2% and 27.3% of patients in the vinorelbine and topotecan trials, respectively. PMID- 12645910 TI - Proteomics versus immunoassays. PMID- 12645911 TI - A personal view of molecular technology and how it has changed biology. PMID- 12645912 TI - The ubiquitin superfamily: members, features, and phylogenies. AB - The ubiquitin superfamily is a rich repository of small, conserved, functionally unique, and important proteins. Its member proteins fold simply and similarly, with kinetic and thermodynamic ease (Sorenson, J. M.; Head-Gordon, T. Toward minimalist models of larger proteins: A ubiquitin-like protein. Proteins 2002, 46, 368-379). They have been implicated in numerous cancers, neurodegenerations, inflammations, and various disorders affecting signal transduction or protein half-life. These proteins serve the cell generally as portable recognition tags with distinct intracellular roles; indeed, tagging with small protein modifiers has become a new hallmark of post-translational modifications and other signal transduction phenomenon (Finley, D. J. Signal transduction. An alternative to destruction. Nature 2001, 412, 283, 285-286). Because many ubiquitin-like proteins bear similarities in sequence, structure, and function, we gathered protein sequences containing the ubiquitin domain from public databases and created a highly granular and defined protein catabolism database to catalog, summarize, reference, and relate them to their targets and specific ligases (to be described elsewhere). In this paper, we reveal a compilation of proteins possessing the ubiquitin domain. This comprises the first and most important part of our database content. We searched available organismal proteomes for sequence related members of the ubiquitin superfamily and here present over 200 proteins possessing this domain. These proteins were organized phylogenetically and functionally, thereby defining several new families. To our knowledge, this is the most complete assemblage of ubiquitin domains to date. PMID- 12645913 TI - Protein variability in Meloidogyne spp. (Nematoda:Meloidogynidae) revealed by two dimensional gel electrophoresis and mass spectrometry. AB - Total protein variation as revealed by two-dimensional electrophoresis (2D-E) was studied in 18 isolates from populations of Meloidogyne arenaria (six isolates), Meloidogyne incognita (10 isolates), and Meloidogyne javanica (one isolate) plus an unclassified isolate. Gels (80 x 60 x 0.75 mm) were silverstained and digitized in order to compare their protein patterns. Optical density and position of protein patterns were measured using statistical cluster analysis and computer-assisted image analysis software. Only those protein stains or positions that were clearly defined (i.e., without background) were considered. The number of positions in gels ranged from 86 to 203. Each of these positions had 95 clearly expressed proteins that were present in at least two replicates for each isolate. Spot position was considered a taxonomical character with two different states: presence (1) and absence (0). Accordingly, genetic distance was estimated among isolates and species, and a phylogenetic tree was constructed following the cladistic approach based on maximum parsimony analysis. Isolates of M. arenaria- M. javanica--Meloidogyne sp. and of M. incognita formed two separate monophyletic groups. Both groups were clearly defined on the basis of two sets of protein positions that can be considered as diagnostic characters. An attempt to identify these proteins by mass spectrometry was made. Group diagnostic proteins for M. incognita and M. arenaria (and for other proteins common to all isolates) were distinguished by protonated mass signals in the MALDI fingerprinting spectrum. PMID- 12645914 TI - Bioinformatical analysis of G-protein-coupled receptors. AB - G-protein-coupled receptors play a key role in cellular signaling networks that regulate various physiological processes, such as vision, smell, taste, neurotransmission, secretion, inflammatory, immune responses, cellular metabolism, and cellular growth. These proteins are very important for understanding human physiology and disease. Many efforts in pharmaceutical research have been aimed at understanding their structure and function. Unfortunately, because they are difficult to crystallize and most of them will not dissolve in normal solvents, so far very few G-protein-coupled receptor structures have been determined. In contrast, more than 1000 G-protein-coupled receptor sequences are known, and many more are expected to become known soon. In view of the extremely unbalanced state, it would be very useful to develop a fast sequence-based method to identify their different types. This would no doubt have practical value for both basic research and drug discovery because the function or binding specificity of a G-protein coupled receptor is determined by the particular type it belongs to. To realize this, a statistical analysis has been performed for 566 G-protein-coupled receptors classified into seven different types. The results indicate that the types of G-protein-coupled receptors are predictable to a considerable accurate extent if a good training data set can be established for such a goal. PMID- 12645915 TI - Differential protein expression in the cytosol fraction of an MCF-7 breast cancer cell line selected for resistance toward melphalan. AB - Analysis of differential protein expression in the cytosol of melphalan-resistant and -susceptible MCF-7 cell lines has been carried out using a combination of two dimensional gel electrophoresis, mass spectrometry, and bioinformatics. Comparison of multiple digitized gel arrays detected several spots as candidates for differentially expressed proteins in melphalan-resistant MCF-7 cells. The up regulated proteins included retinoic acid binding protein II, an isoform of the macrophage migration inhibition factor, and other unidentified proteins. The down regulated proteins included calreticulin, cyclophin A, and an isoform of the 27 kD heat shock protein. Correlation of the differential expression of some of the proteins with acquired resistance of MCF7 cells to melphalan is discussed. PMID- 12645916 TI - An isotope coding strategy for proteomics involving both amine and carboxyl group labeling. AB - This paper describes a heavy isotope coding strategy for the analysis of all types of tryptic peptides, including those that are N-terminally blocked and from the C-terminus of proteins. The method exploits differential derivatization of amine and carboxyl groups generated during proteolysis as a means of coding. Carboxyl groups produced during proteolysis incorporate 18O from H218O. Peptides from the C-terminus of proteins were not labeled with 18O unless they contained a basic C-terminal amino acid. Primary amines from control and experimental samples were differentially acylated after proteolysis with either 1H3- or 2H3-N acetoxysuccinamide. When these two types of labeling were combined, unique coding patterns were achieved for peptides arising from the C-termini and blocked N termini of proteins. This method was used to (1) distinguish C-terminal peptides in model proteins, (2) recognize N-terminal peptides from proteins in which the amino terminus is acylated, and (3) identify primary structure variations between proteins from different sources. PMID- 12645917 TI - An alternative strategy to determine the mitochondrial proteome using sucrose gradient fractionation and 1D PAGE on highly purified human heart mitochondria. AB - An alternative strategy for mitochondrial proteomics is described that is complementary to previous investigations using 2D PAGE techniques. The strategy involves (a) obtaining highly purified preparations of human heart mitochondria using metrizamide gradients to remove cytosolic and other subcellular contaminant proteins; (b) separation of mitochondrial protein complexes using sucrose density gradients after solubilization with n-dodecyl-beta-D-maltoside; (c) 1D electrophoresis of the sucrose gradient fractions; (d) high-throughput proteomics using robotic gel band excision, in-gel digestion, MALDI target spotting and automated spectral acquisition; and (e) protein identification from mixtures of tryptic peptides by high-precision peptide mass fingerprinting. Using this approach, we rapidly identified 82 bona fide or potential mitochondrial proteins, 40 of which have not been previously reported using 2D PAGE techniques. These proteins include small complex I and complex IV subunits, as well as very basic and hydrophobic transmembrane proteins such as the adenine nucleotide translocase that are not recovered in 2D gels. The technique described here should also be useful for the identification of new protein-protein associations as exemplified by the validation of a recently discovered complex that involves proteins belonging to the prohibitin family. PMID- 12645918 TI - Targeted proteomics of low-level proteins in human plasma by LC/MSn: using human growth hormone as a model system. AB - This paper describes the profiling of human growth hormone (hGH) in human plasma in order to assess the dynamic range of the ion-trap mass spectrometer for proteomic studies of complex biological samples. Human growth hormone is an example of a low-level plasma protein in vivo, present at subfemtomole levels. This study was performed on a plasma sample in which hGH has been spiked at 10 fold above the natural level, that is approximately 16 pg/microL of plasma. Initially, the measurement was carried out without any sample enrichment and consisted of the following steps: the full set of plasma proteins were reduced, alkylated, and digested with trypsin, and the resulting peptides were separated on a capillary C-18 column and then detected by ion-trap mass spectrometry (1D LC/MS). In addition, this study provided a global view of the serum proteome with over 200 plasma proteins being preliminarily identified. In the MS/MS analysis, hGH was detected by characterization of the first tryptic peptide (T1). The initial identification was confirmed by alternative approaches, which also allowed the evaluation of different sample purification protocols. First, the plasma sample containing hGH was fractionated on a reversed-phase HPLC column and digested, and hGH could now be identified by MS/MS measurements of two tryptic peptides (T1 and T4) by the same 1D LC/MS protocol. In addition, the assignment of peptide identity was made with higher certainty (as measured by an algorithm score). The plasma sample was also fractionated by 1D and 2D gel electrophoresis, the selected bands were digested and analyzed again by the 1D LC/MS protocol. In both cases using the gel prepurifications, hGH was identified with additional peptides. Finally, the plasma sample was analyzed by 2D chromatography (ion exchange and reversed phase) on a new instrumental platform (ProteomeX), and hGH was identified by the observation of five tryptic peptides. In conclusion, these experiments were able to detect growth hormone in the low femtomole level with a dynamic range of 1 in 40 000 by several independent approaches. The amount of growth hormone, while 10-fold above normal in vivo levels, represents concentrations that may be present in disease states (such as acromegaly) and also in doping control measurements. These studies have demonstrated that shotgun sequencing approaches (LC/MS/MS) not only can profile high-abundance proteins in complex biological fluids but also have the potential to identify and quantitate low-level proteins present in such complex mixtures without extensive prepurification protocols. A key to such studies, however, is to use targeted approaches that reduce the complexity of the solute mixture that is presented to the mass spectrometer at a given time point. The various sample preparation protocols described here all improved the quality of the hGH measurement, although in this study the 2D chromatographic approach gave the greatest sequence coverage. PMID- 12645919 TI - Combining hierarchical clustering and self-organizing maps for exploratory analysis of gene expression patterns. AB - Self-organizing maps (SOM) constitute an alternative to classical clustering methods because of its linear run times and superior performance to deal with noisy data. Nevertheless, the clustering obtained with SOM is dependent on the relative sizes of the clusters. Here, we show how the combination of SOM with hierarchical clustering methods constitutes an excellent tool for exploratory analysis of massive data like DNA microarray expression patterns. PMID- 12645920 TI - A new form of crystalline rubisco and the conversion to its common dodecahedral form. AB - In this paper, we present a new purification procedure that yields a new crystalline form of rubisco and has enabled us to completely remove this most abundant protein from tobacco leaf extract. The crystals formed within 48 h after refrigeration at 4 degrees C at pH 5.6. However, these crystals were not well ordered crystals and lacked well-defined facets or edges. The remaining leaf extract (fraction 2 protein) was void of rubisco. Conversion of this new crystalline form of rubisco to its common dodecahedral form was achieved by dialysing the protein solution in Tris buffer at pH 8.0 or purified water. Since the molecular size of its large subunit of rubisco (55 kD) is similar to that of the papillomavirus capsid protein, L1 (57 kD), its complete removal from fraction 2-protein may facilitate the detection, purification, and recovery of the Li protein. PMID- 12645921 TI - Stressed cells survive better with light. PMID- 12645922 TI - Hormonal, cellular, and molecular control of prostatic development. AB - The prostate is a male accessory sex gland found only in mammals that functions to produce a major fraction of seminal fluid. Interest in understanding the biology of the prostate is driven both by the fascinating nature of the developmental processes that give rise to the prostate and by the high incidence in humans of prostatic diseases, including prostatic adenocarcinoma and benign prostatic hyperplasia. This review summarizes the current state of knowledge of the cellular and molecular processes that control prostatic development. Insight into the mechanisms that control prostatic development has come from experimental embryological work as well as from the study of mice and humans harboring mutations that alter prostatic development. These studies have demonstrated a requirement for androgens throughout prostatic development and have revealed a series of reciprocal paracrine signals between the developing prostatic epithelium and prostatic mesenchyme. Finally, these studies have identified several specific gene products that are required for prostatic development. While research in recent years has greatly enhanced our understanding of the molecular control of prostatic development, known genes cannot yet explain in molecular terms the complex biological interactions that descriptive and experimental embryological studies have elucidated in the control of prostatic development. PMID- 12645924 TI - Mutations in Nop60B, the Drosophila homolog of human dyskeratosis congenita 1, affect the maintenance of the germ-line stem cell lineage during spermatogenesis. AB - Spermatogenesis in Drosophila is maintained by germ-line stem cells. These cells undergo self-renewing divisions and also generate daughter gonial cells, whose function is to amplify the germ cell pool. Gonial cells subsequently differentiate into spermatocytes that undergo meiosis and generate haploid gametes. To elucidate the circuitry that controls progression through spermatogenic stem cell lineages, we are identifying mutations that lead to either excess germ cells or germ cell loss. From a collection of male sterile mutants, we identified P-element-induced hypomorphic alleles of nop60B, a gene encoding a pseudouridine synthase. Although null mutations are lethal, our P element-induced alleles generate viable, but sterile flies, exhibiting severe testicular atrophy. Sterility is reversed by P-element excision, and the atrophy is rescued by a Nop60B transgene, confirming identity of the gene. Using cell type-specific markers, we find that testicular atrophy is due to severe loss of germ cells, including stem cells, but much milder effects on the somatic cells, which are themselves maintained by a stem cell lineage. We show that Nop60B activity is required intrinsically for the maintenance of germ-line stem cells. The relationship of these phenotypes to the human syndrome Dyskeratosis congenita, caused by mutations in a Nop60B homolog, is discussed. PMID- 12645923 TI - Hensen's node gives rise to the ventral midline of the foregut: implications for organizing head and heart development. AB - Patterning of the ventral head has been attributed to various cell populations, including endoderm, mesoderm, and neural crest. Here, we provide evidence that head and heart development may be influenced by a ventral midline endodermal cell population. We show that the ventral midline endoderm of the foregut is generated directly from the extreme rostral portion of Hensen's node, the avian equivalent of the Spemann organizer. The endodermal cells extend caudally in the ventral midline from the prechordal plate during development of the foregut pocket. Thus, the prechordal plate appears as a mesendodermal pivot between the notochord and the ventral foregut midline. The elongating ventral midline endoderm delimits the right and left sides of the ventral foregut endoderm. Cells derived from the midline endoderm are incorporated into the endocardium and myocardium during closure of the foregut pocket and fusion of the bilateral heart primordia. Bilateral ablation of the endoderm flanking the midline at the level of the anterior intestinal portal leads to randomization of heart looping, suggesting that this endoderm is partitioned into right and left domains by the midline endoderm, thus performing a function similar to that of the notochord in maintaining left-right asymmetry. Because of its derivation from the dorsal organizer, its extent from the forebrain through the midline of the developing face and pharynx, and its participation in formation of a single midline heart tube, we propose that the ventral midline endoderm is ideally situated to function as a ventral organizer of the head and heart. PMID- 12645925 TI - Autonomous and nonautonomous functions for Hox/Pbx in branchiomotor neuron development. AB - The vertebrate branchiomotor neurons are organized in a pattern that corresponds with the segments, or rhombomeres, of the developing hindbrain and have identities and behaviors associated with their position along the anterior/posterior axis. These neurons undergo characteristic migrations in the hindbrain and project from stereotyped exit points. We show that lazarus/pbx4, which encodes an essential Hox DNA-binding partner in zebrafish, is required for facial (VIIth cranial nerve) motor neuron migration and for axon pathfinding of trigeminal (Vth cranial nerve) motor axons. We show that lzr/pbx4 is required for Hox paralog group 1 and 2 function, suggesting that Pbx interacts with these proteins. Consistent with this, lzr/pbx4 interacts genetically with hoxb1a to control facial motor neuron migration. Using genetic mosaic analysis, we show that lzr/pbx4 and hoxb1a are primarily required cell-autonomously within the facial motor neurons; however, analysis of a subtle non-cell-autonomous effect indicates that facial motor neuron migration is promoted by interactions amongst the migrating neurons. At the same time, lzr/pbx4 is required non-cell autonomously to control the pathfinding of trigeminal motor axons. Thus, Pbx/Hox can function both cell-autonomously and non-cell-autonomously to direct different aspects of hindbrain motor neuron behavior. PMID- 12645926 TI - Antizyme is a target of sex-lethal in the Drosophila germline and appears to act downstream of hedgehog to regulate sex-lethal and cyclin B. AB - The sex determination master switch, Sex-lethal, has been shown to regulate the mitosis of early germ cells in Drosophila melanogaster. Sex-lethal is an RNA binding protein that regulates splicing and translation of specific targets in the soma, but the germline targets are unknown. In an experiment aimed at identifying targets of Sex-lethal in early germ cells, the RNA encoded by gutfeeling, the Drosophila homolog of Ornithine Decarboxylase Antizyme, was isolated. gutfeeling interacts genetically with Sex-lethal. It is not only a target of Sex-lethal, but also appears to regulate the nuclear entry and overall levels of Sex-lethal in early germ cells. This regulation of Sex-lethal by gutfeeling appears to occur downstream of the Hedgehog signal. We also show that Hedgehog, Gutfeeling, and Sex-lethal function to regulate Cyclin B, providing a link between Sex-lethal and mitosis. PMID- 12645927 TI - Requirement for tumor suppressor Apc in the morphogenesis of anterior and ventral mouse embryo. AB - Tumor suppressor Apc (adenomatous polyposis coli) is implicated in the Wnt signaling pathway that is involved in the early embryonic development and tumorigenesis in vertebrates. While the heterozygous null mutant mice develop intestinal polyps, the homozygous embryos die before gastrulation. To investigate the role of Apc in later embryonic development, we constructed a novel hypomorphic Apc allele whose expression was attenuated by approximately 80%. In the hypomorphic Apc homozygous ES cells, reduction in Apc expression caused beta catenin accumulation and Wnt signaling activation. The homozygous mutant mouse embryos survived 3 days longer than the null mutant embryos. Interestingly, they showed anterior truncation, partial axis duplication, and defective ventral morphogenesis. To determine the tissues where Apc functions for anterior and ventral morphogenesis, we constructed chimeric embryos whose epiblast was derived predominantly from the Apc hypomorphic homozygous cells but the visceral endoderm was from the wild type. Although these chimeric embryos still showed some anterior defects, their ventral morphogenesis was rescued. In addition, marker studies indicated that the axial mesendoderm was also defective in the homozygous embryos. Our results provide genetic evidence that expression of Apc at the normal level is essential for both anterior and ventral development, in the epiblast derivatives and visceral endoderm. PMID- 12645929 TI - Neural crest origin of mammalian Merkel cells. AB - Here, we provide evidence for the neural crest origin of mammalian Merkel cells. Together with nerve terminals, Merkel cells form slowly adapting cutaneous mechanoreceptors that transduce steady indentation in hairy and glabrous skin. We have determined the ontogenetic origin of Merkel cells in Wnt1-cre/R26R compound transgenic mice, in which neural crest cells are marked indelibly. Merkel cells in whiskers and interfollicular locations express the transgene, beta galactosidase, identifying them as neural crest descendants. We thus conclude that murine Merkel cells originate from the neural crest. PMID- 12645928 TI - Drosophila perlecan modulates FGF and hedgehog signals to activate neural stem cell division. AB - Mutations in the Drosophila trol gene cause cell cycle arrest of neuroblasts in the larval brain. Here, we show that trol encodes the Drosophila homolog of Perlecan and regulates neuroblast division by modulating both FGF and Hh signaling. Addition of human FGF-2 to trol mutant brains in culture rescues the trol proliferation phenotype, while addition of a MAPK inhibitor causes cell cycle arrest of the regulated neuroblasts in wildtype brains. Like FGF, Hh activates stem cell division in the larval brain in a Trol-dependent fashion. Coimmunoprecipitation studies are consistent with interactions between Trol and Hh and between mammalian Perlecan and Shh that are not competed with heparin sulfate. Finally, analyses of mutations in trol, hh, and ttv suggest that Trol affects Hh movement. These results indicate that Trol can mediate signaling through both of the FGF and Hedgehog pathways to control the onset of stem cell proliferation in the developing nervous system. PMID- 12645930 TI - Spatial and temporal contribution of somitic myoblasts to avian hind limb muscles. AB - Skeletal muscles of the avian limb are derived from mononucleated myogenic precursor cells (myoblasts) that migrate into the somatopleural mesoderm of the developing limb bud from the ventrolateral dermomyotome of limb adjacent somites. In the present study, we utilized replication-deficient lacZ-encoding retroviruses to elucidate the source of myoblasts for all hind limb muscles in the chick and define the distinct patterns of myoblast distribution within the limb. We also examined, using the same marker, whether the time of migration from the somites into the limb dictates the spatial contribution the myoblasts make to the developing musculature, particularly in relation to the proximodistal and dorsovental axes. Finally, we used these investigations to examine whether the precursors of both primary and secondary myotubes are derived from somitic mesoderm, a presumption, which up until now, has not been demonstrated in vivo. Overall, the results of our studies demonstrate that individual somites have a selective spatial pattern of participation in the development of the avian hind limb musculature and contribute to both primary and secondary myotubes. We also show that both early and later migrating myoblasts can contribute fully to the formation of the appendicular muscles. PMID- 12645931 TI - Formation of the digestive system in zebrafish. I. Liver morphogenesis. AB - Despite the essential functions of the digestive system, much remains to be learned about the cellular and molecular mechanisms responsible for digestive organ morphogenesis and patterning. We introduce a novel zebrafish transgenic line, the gutGFP line, that expresses GFP throughout the digestive system, and use this tool to analyze the development of the liver. Our studies reveal two phases of liver morphogenesis: budding and growth. The budding period, which can be further subdivided into three stages, starts when hepatocytes first aggregate, shortly after 24 h postfertilization (hpf), and ends with the formation of a hepatic duct at 50 hpf. The growth phase immediately follows and is responsible for a dramatic alteration of liver size and shape. We also analyze gene expression in the developing liver and find a correlation between the expression of certain transcription factor genes and the morphologically defined stages of liver budding. To further expand our understanding of budding morphogenesis, we use loss-of-function analyses to investigate factors potentially involved in this process. It had been reported that no tail mutant embryos appear to lack a liver primordium, as assessed by gata6 expression. However, analysis of gutGFP embryos lacking Ntl show that the liver is in fact present. We also find that, in these embryos, the direction of liver budding does not correlate with the direction of intestinal looping, indicating that the left/right behavior of these tissues can be uncoupled. In addition, we use the cloche mutation to analyze the role of endothelial cells in liver morphogenesis, and find that in zebrafish, unlike what has been reported in mouse, endothelial cells do not appear to be necessary for the budding of this organ. PMID- 12645932 TI - SNR1 is an essential subunit in a subset of Drosophila brm complexes, targeting specific functions during development. AB - The snr1 gene of Drosophila melanogaster encodes a conserved component of the multiprotein Brahma (Brm) complex, a counterpart to the SWI/SNF complexes that participate in ATP-dependent chromatin remodeling. Loss-of-function and null mutations in the snr1 gene reveal its essential role in Drosophila development. We identified new mutant alleles and ectopically expressed deleted forms to dissect the specific functions of SNR1. Somatic and germ cell clone analyses confirmed its requirement in a continuous and widespread fashion for proper cell fate determination and oogenesis. Expression of SNR1 transgenes revealed unexpected roles in wing patterning, abdomen development, oogenesis, and sustained adult viability. A widespread distribution of SNR1 and BRM on the salivary gland polytene chromosomes showed that the Brm complex associated with many genes, but not always at transcribed loci, consistent with genetic data suggesting roles in both gene activation and repression. Despite essential Brm complex functions in leg development, genetic and protein localization studies revealed that snr1 was not required or expressed in all tissues dependent on Brm complex activities. Thus, SNR1 is essential for some, but not all Brm functions, and it likely serves as an optional subunit, directing Brm complex activity to specific gene loci or cellular processes. PMID- 12645933 TI - Cadherin-mediated cell adhesion and tissue segregation: qualitative and quantitative determinants. AB - It is widely held that segregation of tissues expressing different cadherins results from cadherin-subtype-specific binding specificities. This belief is based largely upon assays in which cells expressing different cadherin subtypes aggregate separately when shaken in suspension. In various combinations of L cells expressing NCAM, E-, P-, N-, R-, or B-cadherin, coaggregation occurred when shear forces were low or absent but could be selectively inhibited by high shear forces. Cells expressing P- vs E-cadherin coaggregated and then demixed, one population enveloping the other completely. To distinguish whether this demixing was due to differences in cadherin affinities or expression levels, the latter were varied systematically. Cells expressing either cadherin at a lower level became the enveloping layer, as predicted by the Differential Adhesion Hypothesis. However, when cadherin expression levels were equalized, cells expressing P- vs E-cadherin remained intermixed. In this combination, "homocadherin" (E-E; P-P) and "heterocadherin" (E-P) adhesions must therefore be of similar strength. Cells expressing R- vs B-cadherin coaggregated but demixed to produce configurations of incomplete envelopment. This signifies that R- to B cadherin adhesions must be weaker than either "homocadherin" adhesion. Together, cadherin quantity and affinity control tissue segregation and assembly through specification of the relative intensities of mature cell-cell adhesions. PMID- 12645934 TI - Disparate peptide-dependent thymic selection outcomes in beta2M-deficient mice versus TAP-1-deficient mice: implications for repertoire formation. AB - Fetal thymic organ cultures of N15-transgenic RAG-2-/- H-2b mice on normal, beta 2 microglobulin (beta2M)-/- or transporter associated with antigen processing (rAP-1)-/- MHCl-deficient backgrounds were used to examine differentiation of thymocytes bearing a TCR specific for a viral peptide bound to H-2Kb. Strong agonists mediate negative selection in all mice whereas weak agonists are positively selecting in beta2MW-/- mice but negatively selecting on TAP-1-/- or normal backgrounds. Very weak agonists and very weak antagonists are generally without effect in beta2M-/- mice yet foster differentiation in TAP-1-/- animals. The 20-40-fold reduction in beta2M4-/- thymic H-2Kb surface expression suggests that the avidity of the TCR for peptide-MHCI accounts for these differences, consistent with effects of TCR density and individual thymic-peptide abundance in peptide-MHC complexes. TCR-self-MHC interaction dominates Kb-based selection, subtly modulated by peptides as revealed by X-ray crystallography. PMID- 12645935 TI - A novel ligand for the NKG2D receptor activates NK cells and macrophages and induces tumor immunity. AB - NK cells are involved in the immune response against viral and microbial infections and tumors. In contrast to B and T cells, NK cells employ various modes of immune recognition. An important mode of immune recognition employed by NK cells is "induced self recognition" exemplified by the NKG2D receptor-ligand system. The NKG2D immunoreceptor, expressed by NK cells, and by activated CD8+ T cells and macrophages, recognizes one of several cell surface ligands that are distantly related to MHC class I molecules (i.e. H60 and Rae1 proteins in mice, and MHC class I chain-related proteins and UL-16-binding proteins in humans). These ligands are not expressed abundantly by most normal cells but are up regulated on cells exposed to various forms of cellular insults. Here we report the cloning of another ligand for NKG2D; transcripts of this ligand are found in a wide variety of tissues and in various tumor cells. Cross-linking of NKG2D with the novel ligand potently activated NK cells and macrophages. Tumor cells ectopically expressing the molecule were efficiently rejected by naive mice, and induced strong protective immunity to the parental, ligand-negative tumor cells. PMID- 12645936 TI - ICOS+ Th cells produce distinct cytokines in different mucosal immune responses. AB - T cell activation, differentiation and effector functions depend on signals delivered through the antigen-specific TCR and non-clonal costimulatory receptors on the T cell. Activated T cells express the inducible costimulator (ICOS). We examined the co-expression of ICOS with Th cytokines in mucosal immune responses. ICOS+CD4+ Th cells expressed strikingly different cytokines depending on the type of infection encountered and the cells' anatomical localization. In the Th2 dominated response to Schistosoma mansoni, ICOS expression of CD4+ cells isolated from the liver was strongly associated with the expression of IL-5, IL-10, IL-13, and T1/ST2, but not with the chemokine receptor CXCR5, a pattern consistent with Th2 effector cells. In the secondary lymphatic organs of schistosome-infected mice, ICOS expression was randomly correlated with Th2 effector-cytokines, but positively correlated with CXCR5 expression; a pattern consistent with follicular Th cells. In Th cells isolated from gut or liver of mice infected with Toxoplasma gondii, ICOS expression was positively correlated with IFN-gamma production. Finally, in the severe combined immunodeficiency transfer colitis model, ICOS expression was strongly positively associated with IFN-gamma and IL-2. Thus, ICOS appears to costimulate distinct effector functions in different immune responses, depending on factors such as the nature of the antigen encountered and localization and chronicity of the immune response. PMID- 12645937 TI - Effect of anti-macrophage inflammatory protein-1alpha on leukocyte trafficking and disease progression in experimental autoimmune uveoretinitis. AB - This study has enabled us to identify the influence of the chemokine, macrophage inflammatory protein-1alpha (MIP-1alpha), on leukocyte behavior at the blood retina barrier in vivo and its link with the inflammatory process and disease pathogenesis. MIP-1alpha has not previously been thought to be effective under conditions of physiological shear flow. However, short-term anti-MIP-1alpha treatment inhibited leukocyte slowing and accumulation and subsequent extravasation of leukocytes at the blood-retina barrier in animals with experimental autoimmune uveoretinitis. This was effective predominantly in the post-capillary venules which have been shown to be the main site of passage of leukocytes across the blood-retina barrier. Long-term anti-MIP-1alpha treatment also prevented decreased leukocyte velocity and reduced disease severity as measured clinically, histologically and in terms of blood-retina barrier breakdown. PMID- 12645938 TI - Downmodulation of antigen presentation by H2-O in B cell lines and primary B lymphocytes. AB - Peptide loading onto MHC class II molecules takes place in endosomal compartments along the endocytic pathway. There, loading is facilitated by the catalytic function of the accessory molecule H2-M, which helps to exchange the invariant chain-derived CLIP peptide in the groove of class II molecules for antigenic peptide. H2-O is another accessory molecule specific to the class II pathway, which is found tightly associated with H2-M and selectively expressed in B cells. Using stable H2-O ribozyme-antisense transfectants, H2-O overexpressing murine B cell lines, and H2-O-transgenic mice, we investigated the effects of H2-O on antigen presentation. The results show that presentation of a variety of exogenous protein antigens to a panel of T cell hybridomas depended on the levels of H2-O in the antigen-presenting B cells. Thus, increased H2-O expression downmodulated, whereas reduced H2-O levels, enhanced presentation. Presentation of endogenous antigen was also diminished by H2-O. Despite the pronounced effects on antigen presentation, the mass spectrometric profiles of peptides eluted from Ab molecules were very similar in cells expressing different H2-O levels. The intracellular location of H2-O inhibitory activity was investigated with the drug chloroquine, which prevents acidification of the endocytic pathway. The observations indicate that H2-O predominantly inhibits antigen presentation in early endosomal compartments. Thus, H2-O appears to skew peptide loading to late endosomal/lysosomal compartments. This may favor presentation of antigens taken up by the B cell receptor. PMID- 12645939 TI - The ABC-transporter signature motif is required for peptide translocation but not peptide binding by TAP. AB - The transporter associated with antigen processing (TAP) translocates peptides from their site of generation in the cytosol to the lumen of the endoplasmic reticulum for binding to MHC class I molecules. TAP is a member of the ATP binding cassette (ABC) transporter family whose members utilize energy from ATP hydrolysis to translocate substrates across membranes. The highly conserved nucleotide-binding domains of ABC transporters couple ATP hydrolysis to substrate translocation by the membrane domains. The conserved 'signature motif' can be identified in the nucleotide-binding domains of all ABC transporters, and may play a role in ATP hydrolysis. Here we show that introduction of mutations into the signature motifs of either TAP1 or TAP2 inhibits the translocation of peptide without affecting binding of either peptide or ATP by TAP. We therefore conclude that the signature motifs in both TAP1 and TAP2 are required after peptide binding to facilitate peptide translocation by TAP. PMID- 12645940 TI - IL-4 regulates IL-12 p40 expression post-transcriptionally as well as via a promoter-based mechanism. AB - IL-12 and IL-4, respectively, dominate Th1 versus Th2 polarization. Additionally IL-4 can inhibit IL-12 p40 production in DC. Here we show that macrophages respond to bacterial CpG-DNA with IL-12 p40 production in an IL-4-sensitive manner. Analysis of the molecular mechanism of this inhibition shows that IL-4 acts by reducing the stability of IL-12 p40 mRNA as well as by affecting promoter activity. IL-4 did not affect early CpG-DNA-induced signal transduction. However, IL-4 reduced the activity of the IL-12 p40 promoter and when de novo transcription of IL-12 p40 mRNA was blocked, IL-4 led to acceleration of IL-12 p40 mRNA degradation. These data show that IL-4 regulates IL-12 p40 expression by influencing promoter activity and by interfering with mRNA stability. PMID- 12645941 TI - Mice transgenic for intracellular interleukin-1 receptor antagonist type 1 are protected from collagen-induced arthritis. AB - Interleukin-1 receptor antagonist (IL-1Ra) is a natural IL-1 inhibitor, which competitively inhibits binding of IL-1 to its receptors. IL-1Ra is produced as four different isoforms, one secreted (sIL-1Ra) and three intracellular (icIL 1Ra1, 2, 3), derived from the same gene. We previously observed increased production of icIL-1Ra1 in the joints of mice with collagen-induced arthritis (CIA). However, due to its intracellular localization, the biological role of icIL-1Ra1 remains unknown. The aim of the present study was to examine the effect of the icIL-1Ra1 isoform, as compared to that of sIL-1Ra, in the CIA model by comparing transgenic (tg) mice overexpressing icIL-1Ra1 or sIL-1Ra to their wild type littermates. Serum levels of tg human IL-1Ra were elevated in sIL-1Ra and, to a lesser extent, also in icIL-1Ra1 mice. Clinical scoring indicated that none of the icIL-1Ra1 or siL-1Ra tg mice developed CIA, whereas arthritis was present in, respectively, 60% and 100% of their wild-type littermates. Histological and radiological analyses confirmed the absence of arthritis in icIL-1Ra1 and sIL-1Ra tg mice. Accordingly, circulating levels of the acute-phase protein serum amyloid A tended to be lower in icIL-1Ra1 tg mice than in their wild-type littermates and were significantly lower in sIL-1Ra tg mice than in controls. In contrast, no difference was observed between the groups regarding serum levels of anti-type II collagen antibodies and ex vivo spleen cell proliferative response to collagen. In conclusion, icIL-1Ra1, which is released into the extracellular space when produced in high amounts, has a similar anti-arthritic effect as sIL-1Ra. PMID- 12645942 TI - Normal B-1 cell development but defective BCR signaling in Lck-/- mice. AB - Mature B cells are grouped into two major subsets, B-1 and B-2, believed to derive from separate lineages. We have recently shown that B-1 cells, which are characterized by CD5 surface expression, specifically exhibit significant levels of the tyrosine kinase Lck in man. Here we show that also in mice Lck expression is restricted to B-1 cells and address the potential role of Lck in B-1 cell development and activation. Using as a model an Lck-/- mouse, we show that, while dispensable for B-1 cell development, Lck is required for full and sustained activation of the tyrosine phosphorylation and MAP kinase cascades triggered by the BCR in CD5+, B-1 cells. The data suggest a potential role for Lck in the achievement of the higher activation threshold required for productive BCR signaling in B-1 as compared to B-2 cells. PMID- 12645943 TI - Costimulation of human CD28- T cells by 4-1BB ligand. AB - The T cell surface protein CD28 provides a critical costimulatory signal for T cell activation. With age, humans accumulate increasing numbers of CD28- T cells, and this loss of CD28 expression is exacerbated certain disease states, such as HIV infection, autoimmune conditions or cancer. It is unclear whether CD28- T cells represent terminally differentiated effector cells or whether they remain sensitive to costimulation by CD28-independent pathways. Here, we demonstrate that 4-1 BB ligand can costimulate human CD28- T cells, resulting in cell division, inflammatory cytokine production, increased perforin levels, enhancement of cytolytic effector function, as well as the up-regulation of the anti-apoptotic protein Bcl-X(L). Thus, human CD28- T cells can respond to costimulatory signals and as such become attractive targets for therapeutic intervention, particularly in chronic infectious and inflammatory diseases where large numbers of these cells accumulate. PMID- 12645945 TI - Biochemical and functional characterization of the interaction between pentraxin 3 and C1q. AB - Pentraxin 3 (PTX3) is a recently characterized member of the pentraxin family of acute-phase proteins produced during inflammation. Classical short pentraxins, C reactive protein, and serum amyloid P component can bind to C1q and thereby activate the classical complement pathway. Since PTX3 can also bind C1q, the present study was designed to define the interaction between PTX3 and C1q and to examine the functional consequences of this interaction. A dose-dependent binding of both C1q and the C1 complex to PTX3 was observed. Experiments with recombinant globular head domains of human C1q A, B, and C chains indicated that C1q interacts with PTX3 via its globular head region. Binding of C1q to immobilized PTX3 induced activation of the classical complement pathway as assessed by C4 deposition. Furthermore, PTX3 enhanced C1q binding and complement activation on apoptotic cells. However, in the fluid-phase, pre-incubation of PTX3 with C1q resulted in inhibition of complement activation by blocking the interaction of C1q with immunoglobulins. These results indicate that PTX3 can both inhibit and activate the classical complement pathway by binding C1q, depending on the way it is presented. PTX3 may therefore be involved in the regulation of the innate immune response. PMID- 12645944 TI - T cell costimulation by the hepatitis C virus envelope protein E2 binding to CD81 is mediated by Lck. AB - Binding of the hepatitis C virus (HCV) envelope protein E2 to CD81 provides a costimulatory signal for human T cells. This phenomenon may play a role in liver damage and autoimmune manifestations associated with HCV infection. Here we show that cross-linking of CD81 by HCV E2 induced a calcium flux in T cells that depends on Lck since it was blocked by PP1 and absent in Lck-deficient Jurkat T cells. In wild-type Jurkat cells, Lck was activated by CD81 cross-linking, and CD81, like Lck, was found in lipid rafts. Indeed, the integrity of the raft compartment was required for the induction of a calcium flux by E2, since methyl beta-cyclodextrin abolished this response. A requirement for TCR/CD3 expression was indicated by the absence of a calcium flux following E2 stimulation of TCR/CD3-deficient Jurkat cells. CD81 cross-linking increased and prolonged the anti-CD3-induced tyrosine phosphorylation of TCR1 and of other proteins, indicating that the CD81-mediated signal converges with the TCR/CD3 signaling cascade at its most upstream step. In conclusion, we propose that the costimulatory effects of HCV E2 on T cells depend on CD81 cross-linking that activates Lck through raft aggregation and thus leads to enhanced TCR signaling. PMID- 12645946 TI - Kinetics and expression patterns of chemokine receptors in human CD4+ T lymphocytes primed by myeloid or plasmacytoid dendritic cells. AB - We investigated the kinetics of expression of 12 chemoattractant receptors as a function of cell division following priming of human naive CD4+ T cells by different populations of dendritic cells (DC) and under conditions favoring Th1 or Th2 differentiation. Two chemokine receptors, CXCR3 and CXCR5, were rapidly up regulated following T cell activation by either monocyte-derived DC, myeloid DC (mDC) or plasmacytoid DC (pDC). While CXCR5 expression was transient, expression of CXCR3 at advanced cell divisions was dependent on differentiation, being expressed at high levels on Th1 cells. Several other receptors (CCR2, CCR3, CCR4, CCR5, CXCR6 and CRTh2) were acquired progressively as a function of cell division and in a fashion that was influenced by polarizing cytokines. The Th2-associated chemoattractant receptors CRTh2 and CCR3 were up-regulated with slower kinetics compared to the Th1-associated receptors CXCR3 and CXCR6, consistent with a different kinetics and efficiency of polarization. Moreover, CCR4 and CXCR6 were preferentially induced in T cells activated by mDC and pDC, respectively. Finally, CXCR5 and CCR7 were also rapidly and transiently up-regulated in memory T cells following TCR stimulation. These results indicate a complex chemokine receptor regulation dependent on both T cell activation and differentiation state. In addition, they reveal the existence of DC-specific cues for the regulation of T cell migratory capacity. PMID- 12645947 TI - Involvement of macrophage mannose receptor in the binding and transmission of HIV by macrophages. AB - The human immunodeficiency virus (HIV) is an enveloped virus whose surface glycoprotein gp120 binds CD4 on target cell membranes to initiate infection. About half of the carbohydrates on gp120 are terminally mannosylated, a pattern common to many pathogens. We have examined the ability of macrophage mannose receptor (MMR) on primary monocyte-derived macrophages to bind HIV and facilitate its transmission to T cells. We adapted the tyramide signal amplification system for fluorescence detection of HIV bound to macrophages. Our data show that approximately 60% of the initial association of HIV with macrophages that lack expression of DC-SIGN (a dendritic cell-specific ICAM-3 receptor/HIV-1-binding protein) is MMR mediated, as evidenced by inhibition with mannan, D-mannose, EDTA, and soluble mannose-binding lectin, but not by D-galactose. This inhibition is not seen in cells that lack MMR. Macrophages are able to mediate transmission of bound HIV to co-cultured T cells, and this transmission is blocked up to 80% by inhibitors of MMR binding. Unlike virus bound to DC-SIGN, macrophage-bound HIV has a slightly lower half-life compared to free virus, with no transmission in co culture observed beyond 24 h after virus binding to macrophages. Results obtained with endocytosis inhibitors indicate that this decrease in viral longevity is due to rapid internalization of macrophage-bound HIV. Together, these results suggest a substantial role for MMR in the binding and transmission of HIV-1 by macrophages. PMID- 12645948 TI - CCR8-dependent activation of the RAS/MAPK pathway mediates anti-apoptotic activity of I-309/ CCL1 and vMIP-I. AB - We have previously shown that the CC-chemokine 1-309 (CCL1) protects mouse thymic lymphomas against corticoid-induced apoptosis. Here, we analyzed the signal transduction pathways involved in this activity on BW5147 lymphoma. Inhibition of the CCL1 activity by pertussis toxin suggested the involvement of a G protein coupled chemokine receptor. The role of CCR8 was supported by the observation that vMIP-I, another CCR8-ligand identified from the genome of a T cell transforming herpes virus, shared CCL1 anti-apoptotic activity. In addition to CCR8, BW5147 cells also expressed the CXCR4 receptor but its ligand, SDF-1 (CXCL12) showed only a modest anti-apoptotic activity. Other chemokines acting on CCR2, CCR4 and CCR5 failed to protect against apoptosis and to induce BW5147 chemotaxis, suggesting that these receptors were not functionally expressed. By contrast, both CCL1 and vMIP-I up-regulated ERK1/2 MAPK phosphorylation in BW5147 cells. Further analysis demonstrated that CCL1 activates the MAPK pathway in CCR8 transfected CHO cells. The implication of this pathway was confirmed by the fact that PD98059, an inhibitor of MEK kinases, as well as a dominant negative isoform of the M-RAS protein specifically blocked the anti-apoptotic activity of CCL1. PMID- 12645949 TI - High antigen dose and activated dendritic cells enable Th cells to escape regulatory T cell-mediated suppression in vitro. AB - CD4+CD25+ regulatory T cells (Tregs) are critical for peripheral tolerance and prevention of autoimmunity. In vitro coculture studies have revealed that increased costimulation breaks Treg-mediated suppression in response to anti-CD3 or antigen. However, it was unclear whether loss of suppression arose from inactivation of Tregs or whether increased stimulation caused Th cells to escape suppression. We have investigated conditions that allow or override Treg-mediated suppression using DO11.10 TCR-transgenic T cells and chicken ovalbumin peptide 323-339-pulsed antigen-presenting cells. Treg suppression of Th proliferation is broken with potent stimulation, using activated spleen cells and high antigen dose, but is intact at low antigen dose. Costimulation with CD80 and CD86 expressed on activated dendritic cells was essential for Th cell escape from suppression at a high antigen dose. Potently stimulated Tregs were functional since they reduced levels of IL-2, IFN-gamma, IL-4 and Th CD25 expression in cocultures. Furthermore, Tregs responding to high antigen dose and activated splenocytes retained the ability to suppress proliferation, but only of Th cells responding to a sub-optimal dose of independent antigen. Together, our results demonstrate that under conditions of strong antigen-specific stimulation, Tregs remain functional, but Th cells escape Treg-mediated suppression. PMID- 12645950 TI - Estradiol enhances primary antigen-specific CD4 T cell responses and Th1 development in vivo. Essential role of estrogen receptor alpha expression in hematopoietic cells. AB - It is widely accepted that females have superior immune responses than males, but the ways by which sex hormones may enhance T cell responses are still poorly understood. In the present study, we analyzed the effect of estrogens on CD4 T cell activation and differentiation after immunization with exogenous antigens. We show that administration of low doses of 17beta-estradiol (E2) to castrated female mice results in a striking increase of antigen-specific CD4 T cell responses and in the selective development of IFN-gamma-producing cells. Quantitative assessment of the frequency of T cells bearing a public TCR beta chain CDR3 motif demonstrated that the clonal size of primary antigen-specific CD4 T cells was dramatically increased in immune lymph nodes from E2-treated mice. By using mice with disrupted estrogen receptor (ER) alpha or beta genes, we show that ERalpha, but not ERbeta, was necessary for the enhanced E2-driven Th1 cell responsiveness. Furthermore, ERalpha expression in hematopoietic cells was essential, since E2 effects on Th1 responses were only observed in mice reconstituted with bone marrow cells from ERalpha+/+, but not ERalpha-deficient mice. These results demonstrate that estrogen administration promotes strong antigen-specific Th1 cell responses in a mechanism that requires functional expression of ERalpha in hematopoietic cells. PMID- 12645951 TI - Antigen-presenting cell exosomes are protected from complement-mediated lysis by expression of CD55 and CD59. AB - Exosomes are secreted nanometer-sized vesicles derived from antigen-presenting cells, which have attracted recent interest as they likely play important roles in immune regulation, and their use as cell-free tools for immunotherapy has been proposed. Liposomes used clinically as transport vehicles can activate the complement system, leading to their rapid degradation and significant inflammatory toxicity. The use of isolated exosomes in therapy, therefore, may also elicit complement activation, reducing their potential efficacy. We have examined the expression and functional roles of the membrane regulators of complement (CD46, CD55 and CD59) on antigen-presenting cell-derived exosomes. Exosomes express the glycosylphosphatidylinositol (GPI)-anchored regulators CD55 and CD59, but not the transmembrane protein CD46. Antibody blocking of CD55 in the presence of sensitizing antibody (w6/32) and human serum resulted in increased C3b deposition and significantly increased exosome lysis. Blockade of CD59 also resulted in significant lysis, while blocking both CD55 and CD59 increased lysis still further. We conclude that exosomes express GPI-anchored complement regulators in order to permit their survival in the extracellular environment. PMID- 12645952 TI - The C-type lectin DC-SIGN (CD209) is an antigen-uptake receptor for Candida albicans on dendritic cells. AB - Dendritic cells (DC) that express the type II C-type lectin DC-SIGN (CD209) are located in the submucosa of tissues, where they mediate HIV-1 entry. Interestingly, the pathogen Candida albicans, the major cause of hospital acquired fungal infections, penetrates at similar submucosal sites. Here we demonstrate that DC-SIGN is able to bind C. albicans both in DC-SIGN-transfected cell lines and in human monocyte-derived DC. The binding was shown to be time- as well as concentration-dependent, and live as well as heat-inactivated C. albicans were bound to the same extent. Moreover, in immature DC, DC-SIGN was able to internalize C. albicans in specific DC-SIGN-enriched vesicles, distinct from those containing the mannose receptor, the other known C. albicans receptor expressed by DC. Together, these results demonstrate that DC-SIGN is an exquisite pathogen-uptake receptor that captures not only viruses but also fungi. PMID- 12645953 TI - Characterization of H+,K+-ATPase T cell epitopes in human autoimmune gastritis. AB - Human autoimmune gastritis (AIG) is an organ-specific inflammatory disorder leading to gastric atrophy and pernicious anemia. Gastric H+,K(+)-ATPase was identified as the autoantigen in both human disease and experimental murine AIG (EAIG). Studies of EAIG significantly contributed to current knowledge of human AIG, but to what extent EAIG mimics AIG is still debated, and the autoantigenic epitopes in AIG are yet unknown. This study aimed to identify the H+,K(+)-ATPase epitopes recognized by gastric T cell clones from AIG patients, to define their TCR Vbeta usage and epitope-induced cytokine response. Sixteen H+,K(+)-ATPase reactive CD4+ gastric T cell clones of four AIG patients were tested for proliferation to overlapping 15-mer peptides spanning the a and beta chains of H+,K(+)-ATPase. We identified 6 epitopes in the a chain and 5 in the beta chain; TCR Vbeta usage was not restricted. Four (36%) of the 11 H+,K(+)-ATPase epitopes recognized in AIG were found to overlap with epitopes that are relevant in EAIG, including a previously described gastritogenic epitope. Gastric T cell recognition of the peptide epitopes resulted in secretion of Th1 cytokines. Our data suggest a striking similarity between human AIG and EAIG, at the epitope level, with regard to cytokine secretion and likely also with regard to pathogenic mechanisms. PMID- 12645954 TI - Deviation of islet autoreactivity to cryptic epitopes protects NOD mice from diabetes. AB - To better understand loss of self-tolerance in diabetes-prone NOD mice, we are generating ICA69 transgenes under control of the tetracycline-regulated tet07 minimal promoter. In vitro pilot studies showed leaky transgene expression, but addition of beta-globin genomic insulator flanks prevented leakage and dramatically enhanced transgene expression even in transient transfection, with excellent suppression by Doxycycline. In vivo, the accidental loss of insulator flanks during transgene insertion in one transgenic NOD founder, tet1, re established leakiness with high level, exclusive ICA69-transgene expression in stromal elements of thymus and spleen. This led to persistent deletion of T cells targeting the immunodominant ICA69 epitope, Tep69, but emergence of T cell pools targeting cryptic ICA69 epitopes not normally generated in sufficient density to select and maintain ICA69-autoreactive T cells. This subtle modification of T cell repertoires reduced insulitis, and protected from diabetes in transgenics and in wild-type mice carrying irradiated tet1 thymus grafts. The low pathogenicity of T cells targeting cryptic epitopes likely reflects the fact that the major ICA69 determinant presented in the islet milieu remains Tep69, while cryptic epitopes are under-represented. Deviation of T cell autoreactivity from major to cryptic target epitopes in tet1 mice provides a fortuitous model to explain previously observed diabetes protection by immunotherapy or autoantigen transgenes despite apparent failure to achieve tolerance to the full length islet antigens. PMID- 12645955 TI - Adoptive transfer of an anti-MART-1(27-35)-specific CD8+ T cell clone leads to immunoselection of human melanoma antigen-loss variants in SCID mice. AB - The identification of appropriate mouse models could be useful in carefully evaluating the actual role of the in vivo development of antigen-loss variants during antigen-specific vaccine therapy of human tumors. In this study we investigated the level of efficacy of a MART-1/Melan-A-specific CD8+ T cell clone against its autologous melanoma in a severe combined immunodeficiency (SCID) mouse model, in which the tumor cells expressed in vivo heterogeneous and suboptimal levels of MART-1. The subcutaneous co-injection of the MART-1/Melan-A reactive T cell clone A42 with MART-1/Melan-A+ autologous human melanoma cells into SCID mice caused a total inhibition of tumor growth. However, the systemic treatment with A42 clone lymphocytes resulted in only 50-60% inhibition of tumor growth, although the T cell clone targeted the tumors and the MART-1+ cells virtually disappeared from the tumors. This study suggests that an immunotherapy based on the expansion of an antigen-specific T cell clone generated in vitro is highly efficient in abolishing tumor growth when the target antigen is fully expressed, but leads to in vivo immunoselection of antigen-loss variants in the presence of suboptimal levels of antigen expression. Furthermore, this work shows that human tumors/SCID mouse models may be useful in evaluating the in vivo efficacy of adoptive immunotherapies. PMID- 12645956 TI - The human TREM gene cluster at 6p21.1 encodes both activating and inhibitory single IgV domain receptors and includes NKp44. AB - We have characterized a cluster of single immunoglobulin variable (IgV) domain receptors centromeric of the major histocompatibility complex (MHC) on human chromosome 6. In addition to triggering receptor expressed on myeloid cells (TREM)-1 and TREM2, the cluster contains NKp44, a triggering receptor whose expression is limited to NK cells. We identified three new related genes and two gene fragments within a cluster of approximately 200 kb. Two of the three new genes lack charged residues in their transmembrane domain tails. Further, one of the genes contains two potential immunotyrosine Inhibitory motifs in its cytoplasmic tail, suggesting that it delivers inhibitory signals. The human and mouse TREM clusters appear to have diverged such that there are unique sequences in each species. Finally, each gene in the TREM cluster was expressed in a different range of cell types. PMID- 12645957 TI - Episodic memory meets working memory in the frontal lobe: functional neuroimaging studies of encoding and retrieval. AB - Recent functional-neuroimaging studies have provided a wealth of new information suggesting that regions of the prefrontal cortex play a role in episodic memory encoding and retrieval. This review seeks to evaluate the results of these studies in the context of one general model that has proposed that the left prefrontal cortex is preferentially involved in episodic memory encoding, whereas the right prefrontal cortex is preferentially involved in episodic memory retrieval, irrespective of the type (e.g., modality) of information being remembered. The origins of this framework are considered in some detail and then all relevant functional-neuroimaging studies are critically reviewed. The results of this review fail to provide support for the functional-asymmetry model, suggesting instead that episodic memory encoding and retrieval may actually involve similar regions of the lateral prefrontal cortex when all factors relating to the type of stimulus material (i.e., modality), are appropriately controlled. PMID- 12645958 TI - Neural origins of the P300. AB - A review of the literature investigating the neural origins of detection behavior in humans reveals two event-related potential components, P3a and P3b, each with a distinct neural organization and cognitive function.The P3a is involved in automatic novelty detection and characterized by a more anterior cortical distribution, whereas the P3b is concerned with volitional target detection and has a more posterior cortical distribution. Intracranial investigation, studies with patients with focal brain lesions, and functional neuroimaging (fMRI) studies converge with scalp-recorded event-related potential (ERP) data in suggesting that a widespread cortical network gives rise to both automatic and controlled detection behavior. The main regions consistently attributed to generating detection-related brain activation include the temporal-parietal junction, medial temporal complex, and the lateral prefrontal cortex. The extant human and animal literature addressing the neural networks, neuropharmacological underpinnings, and behavioral significance of the P300 potential will be reviewed. PMID- 12645959 TI - Sleep after immobilization stress and sleep deprivation: common features and theoretical integration. AB - The goal of the present paper is to elucidate and to resolve contradictions in the relationships among different forms of stress, sleep deprivation, and paradoxical sleep (PS) functions. Acute immobilization stress and the stress of learned helplessness are accompanied by an increase of PS, whereas the stress of defense behavior and the stress of self-stimulation cause PS reduction. Recovery sleep after total sleep deprivation performed on the rotating platform is marked by a dramatic rebound of PS although NREM (non-rapid eye movement) sleep deprivation is more prominent than PS deprivation. This PS rebound leads to a quick reversal of the pathology caused by prolonged sleep deprivation. The search activity (SA) concept presents an explanation for these contradictions. SA increases body resistance to stress and diseases, whereas renunciation of search (giving up, helplessness) decreases body resistance. PS and dreams contain covert SA, which compensates for the lack of the overt SA in the preceding period of wakefulness. The requirement for PS increases after giving up and decreases after active defense behavior and self-stimulation. Immobilization stress prevents SA in waking behavior and increases the need in PS. Sleep deprivation on the rotating platform, like immobilization stress, prevents SA, produces conditions for learned helplessness and, suppresses PS. Such a combination increases PS pressure and decreases body resistance. PMID- 12645960 TI - Acute arsenic poisoning treated by intravenous dimercaptosuccinic acid (DMSA) and combined extrarenal epuration techniques. AB - Arsenic poisoning was diagnosed in a 26-year-old man who had been criminally intoxicated over the last two weeks preceding admission by the surreptitious oral administration of probably 10 g of arsenic trioxide (As2O3). The patient developed severe manifestations of toxic hepatitis and pancreatitis, and thereafter neurological disorders, respiratory distress, acute renal failure, and cardiovascular disturbances. In addition to supportive therapy, extrarenal elimination techniques and chelating agents were used. Dimercaprol (BAL) and dimercaptosuccinic acid (DMSA or succimer) were used simultaneously as arsenic chelating agents for two days, and thereafter DMSA was used alone. DMSA was administered by intravenous (20 mg/kg/d for five days, then 10 mg/kg/d for six days) and intraperitoneal route. Intravenous DMSA infusion was well tolerated and resulted in an increase in arsenic blood concentration immediately after the infusion. Continuous venovenous hemofiltration combined with hemodialysis, and peritoneal dialysis were proposed to enhance arsenic elimination. It was calculated that over an 11-day period 14.5 mg arsenic were eliminated by the urine, 26.7 mg by hemodialysis, 17.8 mg by peritoneal dialysis, and 7.8 mg by continuous venovenous hemofiltration. These amounts appeared negligible with regard to the probable ingested dose. The patient died on day 26 from the consequences of multiple organ failure, with subarachnoid hemorrhage and generalized infection caused by Aspergillus fumigatus. PMID- 12645961 TI - Australian tiger snake (Notechis scutatus) and mexican coral snake (Micruris species) antivenoms prevent death from United States coral snake (Micrurus fulvius fulvius) venom in a mouse model. AB - BACKGROUND: Wyeth-Ayerst has discontinued production of Antivenin (Micrurus fulvius). Currently, there is no other approved coral snake antivenom available in the United States. METHODS: This study was a randomized, placebo-controlled and blinded determination of the ability of a Mexican Micrurus (coral snake) antivenom and an Australian Notechis (tiger snake) antivenom to prevent lethality from a United States Micrurus fulvius fulvius venom in a mouse model. Venom dosing was based on an LD50 determined for this experiment. Our comparison groups included: (1) M. f. fulvius venom + Micrurus antivenom, (2) M. f. fulvius venom + Notechis antivenom, (3) M. f. fulvius venom + protein control, (4) 0.9% normal saline + protein control, (5) saline + Notechis antivenom, (6) saline + Micrurus antivenom. Venom dose was 5 times the determined LD50. The antivenom amounts were capable of neutralizing 10 times the venom injected (50 times the LD50). RESULTS: The LD50 of M. f. fulvius venom was determined to be 0.85 mg/kg. All mice in both antivenom test groups were protected from lethality for the entire 24-hour observation period. Six of the 7 mice in the venom test group died, with a survival time of 349 +/- 382 minutes (mean +/- s.d.) after the venom injection. All three groups of control mice survived the entire 24-hour observation period. CONCLUSIONS: Mexican Micrurus antivenom and Australian Notechis antivenom provide protection from lethality in mice envenomated with a United States M. f. filvius venom. PMID- 12645962 TI - Characterization of gabapentin overdose using a poison center case series. AB - BACKGROUND: Gabapentin is an anticonvulsant that is being used for an increasing number of off-label indications. The purpose of this study is to document the clinical manifestations and outcomes of gabapentin exposures reported to poison centers. METHODS: A multicenter prospective observational study of all gabapentin exposures reported to three poisoncenters was conducted between 4/1/98 and 4/1/2000. Cases involving gabapentin only were evaluated. RESULTS: There were 20 cases with gabapentin as the sole substance in doses ranging from 50 mg to 35 g. Ten of the 20 cases involved children and adolescents. Clinical effects developed early and resolved within 10 hours in most patients. Seven cases were managed in the home with only observation. Four of these patients remained asymptomatic. Effects reported in the three symptomatic patients were drowsiness (3) and ataxia (1). Thirteen patients were managed in a health care facility. Nine were symptomatic with reported effects of drowsiness (6), dizziness (3), nausea/vomiting (2), tachycardia (2), and hypotension (2). None of the patients were admitted for medical care. CONCLUSION: In this cases series, gabapentin exposures caused no or minimal toxicity. PMID- 12645963 TI - Regional pharmaceutical preparation for biological and chemical terrorism. AB - BACKGROUND: The United States National Office of Domestic Preparedness has determined that the threat of a biological or chemical attack is very real. As an active participant of a 13-county regional task force, one of the roles of the poison center was to determine the pharmaceutical needs of the community in the event of a terrorist action and develop a financially responsible method of acquisition and storage. METHODS: Working with local health officials, an extensive literature review was conducted to identify possible biological and chemical poisons. Treatment recommendations were identified and an estimated amount to treat 5,000 people for 24hrs was determined. Instead of purchasing the medications, a unique solution utilizing a regional pharmacy wholesaler was used. DISCUSSION: An important element in a biological or chemical terrorist event is the availability of the pharmaceuticals and the capability of delivering them rapidly. The poison center is the ideal agency to help coordinate this endeavor since it is familiar with contemporary therapy and will be aware of the number, location, and status of casualties. Based on the expense involved in the purchase and storage of a large quantity of medications, utilizing a local pharmaceutical distribution company is fiscally responsible. Rotation through normal stock and being readily accessible is another benefit. CONCLUSION: The poison center serves a number of roles in the surveillance, recognition, and treatment of biological and chemical terrorism. Assisting in the development, implementation, and procurement of a pharmaceutical cache is yet another role. PMID- 12645964 TI - Methanol contamination of Romanian home-distilled alcohol. PMID- 12645966 TI - Polymorphisms of paraoxonase (PON1) and their significance in clinical toxicology of organophosphates. AB - Paraoxonase (PON1) is an HDL-associated enzyme capable of hydrolyzing multiple substrates, including several organophosphorous insecticides and nerve agents, oxidized lipids, and a number of drugs or pro-drugs. Several polymorphisms in the paraoxonase (PON1) gene have been described, which have been shown to affect either the catalytic efficiency of hydrolysis or the expression level of PON1. This review discusses the relevance of these polymorphisms for modulating sensitivity to organophosphorous compounds. Animal studies characterizing the PON1 polymorphisms have demonstrated the relevance of PON1 in modulating OP toxicity and have indicated the importance of an individual's PON1 status (i.e., genotype and phenotype taken together) rather than genotyping alone. Nevertheless, direct confirmation in humans of the relevance of PON1 status in conferring susceptibility to OP toxicity is still elusive. Recent studies examining the involvement of PON1 status in determining OP susceptibility of Gulf War veterans, sheep dippers, and individuals poisoned with chemical warfare agents represent a step in the right direction, but more studies are needed, with better documentation of both the level of exposure and the consequences of exposure. PMID- 12645965 TI - Copperhead envenomations in the Carolinas. AB - INTRODUCTION: Although the copperhead (Akistrodon contortrix) is responsible for most Crotaline envenomations in the Carolinas, manifestations and treatment are poorly characterized. OBJECTIVE: We sought to describe the clinical course after copperhead bites. METHOD: Structured review of copperhead exposures reported to a regional poison center from 1997-2000. Hospital records were reviewed when available. Phone followup was attempted. RESULTS: A total of 178 cases were identified. Of these 75% were males. The median age was 31 yr (range 2-93). The bite site included hand (52%), foot (36%), leg (7%), and arm (5%). Classification included dry (7%), mild (48%), moderate (39%), and severe (6%). The most common symptom was pain (93%). Local findings included swelling (94%), fang marks (93%), ecchymosis (53%), erythema (37%), bullae (13%), and tissue necrosis (8%). Eleven of 37 patients developed abnormal PT and/or PTT. Two patients bled. Patients were treated at a healthcare facility in 160 cases, with 79 patients admitted. Opioid analgesics were the most common therapy (81%). Equine-derived antivenin was given in 14 cases (range 2-30 vials). Antivenin reactions developed in three. Two patients received blood products. Surgical treatment included debridement (6), grafting (2), digit amputation (1), digit dermotomy (1), and fasciotomy (1). No patients died. In followup, 18 patients reported limb dysfunction ranging from 5 365 days. CONCLUSION: Copperhead bites typically result in mild to moderate envenomation due to local tissue effects. Significant systemic manifestations are rare. Limb dysfunction can be prolonged. PMID- 12645967 TI - Post-mortem toxicology: what the dead can and cannot tell us. AB - The evaluation of postmortem laboratory assays of drugs needs to be performed in a systematic manner. The condition of the body, drug characteristics, matrix and site analysis are factors which need to be considered in the proper interpretation of an autopsy specimen result. PMID- 12645968 TI - Repeated ingestion of 2-butoxyethanol: case report and literature review. AB - BACKGROUND: Ethylene glycol monobutyl ether (2-butoxyethanol) is not commonly associated with significant human poisoning. Exposures are usually through occupational contact and typically involve inhalation injury. Animal studies report severe hemolysis occurring in rats and mice. Rare published human cases give varied descriptions of the clinical course associated with 2-butoxyethanol poisoning including reports of metabolic acidosis, ethylene glycol production, oxaluria, renal failure, and anemia. We report a case of two separate ingestions (80 to 100 grams) of a glass cleaner concentrate containing 22% 2-butoxyethanol, and its primary metabolite butoxyacetic acid. CASE REPORT: An 18-year-old male ingested 360-480 mL of 22% 2-butoxyethanol on two separate occasions. Approximately 10hours after the first ingestion, the patient developed severe CNS depression, metabolic acidosis, hematuria, and mild elevation of hepatic enzymes. He was treated initially with ethanol therapy but continued to deteriorate and was started on hemodialysis. Approximately 10 days after discharge, the patient ingested 480 mL of the same product and received ethanol and hemodialysis within four hours of ingestion. During his second admission the patient did not develop the delayed severe CNS depression or profound metabolic acidosis. Clinically significant hemolytic anemia, oxaluria, ethylene glycol production, and renal failure were not noted in either episode. The patient recovered on both occasions without sequelae. CONCLUSION: Hemodialysis may be an effective treatment intervention for managing severe acute 2-butoxyethanol intoxication, however, further investigation is warranted. PMID- 12645969 TI - Intoxication with foothill camas (Zigadenus paniculatus). AB - Eight adults ingested foothill camas (Zigadenus paniculatus) bulbs in Juab County, Utah, believing them to be nontoxic wild bulbs. All who ingested the bulbs became ill, and three of them required admission for supportive care. All patients had nausea; other findings included vomiting, abdominal pain, dizziness, near syncope, hypotension, and bradycardia. No specific antidote is available for intoxication with Zigadenus species. Supportive care is indicated and atropine may benefit those with sympytomatic bradycardia and hypotension. PMID- 12645970 TI - Transient and reversible parkinsonism after acute organophosphate poisoning. AB - Parkinsonism is a rare complication in patients with organophosphate poisoning. To date there have been two cases of transient parkinsonism after acute and severe cholinergic crisis, both of which were successfully treated using amantadine, an anti-parkinsonism drug. We report on an 81-year-old woman who was admitted for the treatment of acute severe organophosphate poisoning. Although acute cholinergic crisis was treated successfully with large doses of atropine and 2-pyridine aldoxime methiodide (PAM), extrapyramidal manifestations were noticed on hospital day 6. The neurological symptoms worsened, and the diagnosis of parkinsonism was made by a neurologist on hospital day 9. Immediately, biperiden (5mg), an anti-parkinsonism drug, was administered intravenously, and her symptoms markedly improved. From the following day, biperiden (5 mg/day) was given intramuscularly for eight days. Subsequently, neurological symptoms did not relapse, and no drugs were required. Our patient is the third case of parkinsonism developing after an acute severe cholinergic crisis and the first case successfully treated with biperiden. Patients should be carefully observed for the presence of neurological signs in this kind of poisoning. If present, an anti-parkinsonism drug should be considered. PMID- 12645971 TI - Multiple dose-activated charcoal as a cause of acute appendicitis. AB - We presented a case of a 55-year-old woman who intentionally ingested an unknown amount of carbosulfan, a carbamate insecticide. On admission, her clinical findings were coma, pinpoint pupils, hypersalivation, respiratory failure, bradycardia, and hypotension. Hertrachea was intubated after suction of secretions, and atropine was administered intravenously. After gastric lavage, multiple doses of activated charcoal were instilled through the nasogastric tube over five days (total doses of 840 g). On the fourteenth day, she developed right lower quadrant abdominal pain, anorexia, nausea, and vomiting, and she underwent an appendectomy. On pathologic examination of the specimen, particles of activated charcoal were seen within the dilated part of the appendiculer lumen. The patient was discharged from the hospital after antidepressant therapy at the psychiatry clinic. This case documents that multiple doses of activated charcoal may be associated with acute appendicitis. PMID- 12645972 TI - Mercury vapor inhalation from Chinese red (Cinnabar). AB - INTRODUCTION: Acute inhalation of mercury fumes or vapors is a rare but frequently fatal cause of acute lung injury. This report describes a rare cause of mercury inhalation from Chinese red. CASE REPORT: An 87-year-old male inhaled the vapors from heating Chinese red (Cinnabar, mercury sulphide) intended to treat his foot ulceration. He subsequently developed acute lung injury (progressive dyspnea and acute respiratory failure) that was treated with mechanical ventilation. DMPS (2,3-Dimercapto-1-propanesulfonic acid) and penicillamine were used as chelating agents, and methylprednisolone pulse therapy was used to treat his pulmonary disease. Despite being extubated once, the patient eventually died from profound hypoxemia. CONCLUSION: A rare case of mercury intoxication was due to inappropriate use of an alternative medicine, Chinese red. This case serves as a reminder of the toxicity of the noxious gas from this substance and the importance of being familiar with alternative medicines. PMID- 12645973 TI - Cardiotoxicity associated with intentional ziprasidone and bupropion overdose. AB - BACKGROUND: Ziprasidone (Geodon) and bupropion (Wellbutrin) are medications prescribed for mood and behavior disorders. They have apparently safe cardiac safety profiles in both therapeutic and supra-therapeutic doses. CASE REPORT: A 17-year-old male developed a widened QRS and a prolonged QTc interval following an overdose of ziprasidone and bupropion. He required hospital admission for aggressive cardiac monitoring and antidysrhythmic therapy, stabilizing to baseline by 80 hours post-ingestion. CONCLUSIONS: We present a case that underscores the potential cardiotoxicities of these medications. Ziprasidone and bupropion ingestion can be associated with cardiotoxicities that may require several days of aggressive cardiac monitoring and treatment. PMID- 12645974 TI - Baclofen withdrawal following removal of an intrathecal baclofen pump despite oral baclofen replacement. AB - Intrathecal baclofen is used as a muscle relaxant and antispasmodic in cases of spasticity resulting from central nervous system trauma. The baclofen withdrawal syndrome may include hyperthermia, tachycardia, hypertension, seizures, altered mental status, and psychomotor agitation. We report a case in which the removal of a baclofen pump lead tothe development of severe withdrawal symptoms despite oral baclofen replacement therapy. In order to avoid the development of withdrawal, adequate doses of GABA agonist agents should be administered immediately prior to, and following, baclofen pump removal. PMID- 12645976 TI - Demonstration of a linear optical true-time delay device by use of a microelectromechanical mirror array. AB - We present the design and proof-of-concept demonstration of an optical device capable of producing true-time delay(s) (TTD)(s) for phased array antennas. This TTD device uses a free-space approach consisting of a single microelectromechanical systems (MEMS) mirror array in a multiple reflection spherical mirror configuration based on the White cell. Divergence is avoided by periodic refocusing by the mirrors. By using the MEMS mirror to switch between paths of different lengths, time delays are generated. Six different delays in 1 ns increments were demonstrated by using the Texas Instruments Digital Micromirror Device as the switching element. Losses of 1.6 to 5.2 dB per bounce and crosstalk of -27 dB were also measured, both resulting primarily from diffraction from holes in each pixel and the inter-pixel gaps of the MEMS. PMID- 12645975 TI - Intravenous paraquat poisoning-induced multiple organ failure and fatality--a report of two cases. AB - Paraquat poisoning is the most common cause of fatal herbicide intoxication, mostly through oral ingestion. This work reports two cases of death following intravenous paraquat injection. The clinical courses of the two cases were fulminated and fatal. Toxic symptoms and severe organ function impairment developed soon after paraquat injection. Timely treatment with activated charcoal hemoperfusion with pulse steroid and cyclophosphamide was attempted in both cases; however, both cases died within five days owing to multiple organ failure. In cases of intravenous paraquat intoxication, toxic signs develop more quicker than with oral ingestion. The prognosis of intravenous paraquat intoxication is extremely poor. PMID- 12645977 TI - Optical characterization and applications of a dual-cure photopolymerizable system. AB - The optical response of a photopolymerizable formulation consisting of a bisphenol A epoxy acrylate oligomer, a divinyl ether, and a photoinitiator system containing Rose Bengal was studied by recording holographic gratings. This blend is sensitive to blue-green light. Single- and double-exposure volume phase holograms were recorded. In addition to these examples, surface depth measurements were made by means of a holographic contour technique. PMID- 12645978 TI - Phase-only filtering on the three-dimensional Fourier spectrum of color images. AB - We propose what is to our knowledge a novel technique for filtering a two dimensional multichannel color image based on its interpretation as a three dimensional (3D) signal and the application of a 3D Fourier transform. The color image is considered to have the two usual spatial coordinates with the color distribution as the third dimension of the signal. This approach permits the 3D generalization of the usual phase-only filter and phase correlation, and color pattern recognition can be performed by means of a 3D correlation. We analyze two cases: whitening only along the color Fourier spectrum and whitening the whole 3D Fourier spectrum. We study the transformation induced in the color content of the image by the whitening of the color Fourier spectrum and how it affects the correlation distribution. In particular it is demonstrated that the whitening of the 3D Fourier spectrum improves the recognition performance of the correlation. PMID- 12645979 TI - Rotation invariant color pattern recognition by use of a three-dimensional Fourier transform. AB - Recently, the use of three-dimensional correlation for multichannel pattern recognition has been introduced. In this work we propose the use of circular harmonic components with this new technique to obtain invariance under target rotations. The differences between this method and the previous use of circular harmonic filters for multichannel images are discussed. Also the problem of determining the proper center is studied and, to our knowledge, a new and more understandable criterion to locate it is introduced. Some simulation results to verify the successful operation of the method are included. PMID- 12645980 TI - Volume emission rate tomography from a satellite platform. AB - The possibility of retrieving horizontal atmospheric structure from a series of limb images taken aboard a satellite is discussed and a maximum likelihood expectation maximization algorithm is developed. Examples of the retrieval of horizontal structure with this algorithm, for different S/N (signal-to-noise) ratios and different structures, are presented. It is shown that with this algorithm and even in the presence of substantial observational noise, a S/N equal to 10 for a single observation, it is possible to retrieve both horizontal and vertical atmospheric structure. PMID- 12645981 TI - Superresolution by use of code division multiplexing. AB - Traditional methods for superresolution have sacrificed field of view for resolution. These methods multiplexed different parts of signals' spectrum on different carriers, and thus managed to transfer a wider range of frequency, in a manner that is similar to frequency division multiplexing in classical communication. We propose code division multiplexing for such an application, which has been shown to have superior capabilities. To enable such mutiplexing we propose a unique setup that creates an incoherent cosine transform of the image. A theoretical analysis of the setup is obtained and later compared with the empirical results. PMID- 12645982 TI - Design of three-dimensional superresolution filters and limits of axial optical superresolution. AB - Theories to design a three-dimensional superresolution filter (TDSF) for confocal microscopy are proposed that can obtain a globally optimal solution through linear programming. The designed TDSF is proved to be a phase-only element introducing a phase delay of 0 or pi. Five design examples of the TDSF are presented to demonstrate the validity of these theories, Regardless of transverse superresolution, a curve of Seu(Ga+/-) defined as the maximum value of Strehl ratio S under the axial resolving power of Ga+/- is calculated to set the fundamental limits of axial optical superresolution. Finally, what is to our knowledge a novel analytic expression of Seu(Ga+/-) is deduced. PMID- 12645983 TI - Refined fourier-transform method of analysis of full two-dimensional digitized interferograms. AB - A refined Fourier-transform method of analysis of interference patterns is presented. The refinements include a method of automatic background subtraction and a way of treating the problem of heterodyning. The method proves particularly useful for analysis of long sequences of interferograms. PMID- 12645984 TI - Three-dimensional point spread functions of an optical heterodyne scanning image processor. AB - We study the three-dimensional (3-D) imaging properties of an optical heterodyne scanning image processor. The image processor is a two-pupil optical system capable of 3-D imaging coherently or incoherently, depending on the detection scheme used. We derive the imaging properties in terms of the two pupils and then show an important 3-D imaging application in scanning holography by deriving its 3-D point spread functions and compare them with conventional 3-D imaging systems. PMID- 12645985 TI - Code-multiplexed optical scanner. AB - A three-dimensional (3-D) optical-scanning technique is proposed based on spatial optical phase code activation on an input beam. This code-multiplexed optical scanner (C-MOS) relies on holographically stored 3-D beam-forming information. Proof-of-concept C-MOS experimental results by use of a photorefractive crystal as a holographic medium generates eight beams representing a basic 3-D voxel element generated via a binary-code matrix of the Hadamard type. The experiment demonstrates the C-MOS features of no moving parts, beam-forming flexibility, and large centimeter-size apertures. A novel application of the C-MOS as an optical security lock is highlighted. PMID- 12645986 TI - Particle swarm optimization-based approach for optical finite impulse response filter design. AB - This study presents what is to our knowledge a new and efficient method for the design of an optical finite impulse response (FIR) filter by employing a particle swarm optimization technique. With the method proposed, the design of an optical FIR filter, which is able to provide an arbitrary spectrum output based on crystal birefringence, could be implemented with good performance and high efficiency. The design procedure is discussed. A typical example of a green/magenta filter used in a liquid crystal on silicon projection display is included to demonstrate the feasibility and efficiency of this method in this design process as compared with simulated annealing. PMID- 12645987 TI - Secure optical data storage with random phase key codes by use of a configuration of a joint transform correlator. AB - A secure optical storage based on a configuration of a joint transform correlator by use of a photorefractive material is presented. A key code designed through the use of an optimized algorithm so that its Fourier transform has a uniform amplitude distribution and a uniformly random phase distribution is introduced. Original two-dimensional data and the key code are placed side-by-side at the input plane. Both of them are stored in a photorefractive material as a joint power spectrum. The retrieval of the original data can be achieved with the same key code. We can record multiple two-dimensional data in the same crystal by angular multiplexing and/or key code multiplexing. PMID- 12645988 TI - Biased micromechanical cantilever arrays as optical image memory. AB - We show that an array of optically actuated biased cantilevers can work as an optical data storage, able to encode data stored as arrays of optical pixels (images). Each of these optical pixels can, in addition, have a predetermined pixel depth, expressed as a certain number of gray levels. This new optical memory is able to work at a data rate of approximately 7 GB/s for an image with 128 x 128 pixels. PMID- 12645989 TI - Active optical interconnect based on liquid-crystal grating. AB - We propose a transparent plate of a liquid-crystal grating to be used as a light guide for optical interconnection. To this end, we are empowering the connection itself with active functions, such as switching, wavelength division, power adjustment, etc. In experiments, we built a grating based on in-plane switching. It contains vertically-aligned nematic liquid-crystal molecules between a glass plate with a high refractive index (light guide) and another glass plate with a pair of interdigitated electrodes. Entering a TM wave from an edge of the light guide, we have demonstrated that the activation of diffraction and intensity adjustment for the guided light are both possible. Because a TE wave is barely diffracted, the device also exhibits polarization division capability. PMID- 12645990 TI - Split off-specular reflection and surface scattering from woven materials. AB - We measured radiance distributions for black lining cloth and copper gauze using the convenient technique of wrapping the materials around a circular cylinder, irradiating it with a parallel light source and collecting the scattered radiance by a digital camera. One family of parallel threads (weave or weft) was parallel to the cylinder generator. The most salient features for such glossy plane weaves are a splitting up of the reflection peak due to the wavy variations in local slopes of the threads around the cylinders and a surface scattering lobe due to the threads that run along the cylinder. These scattering characteristics are quite different from the (off-)specular peaks and lobes that were found before for random rough specular surfaces. The split off-specular reflection is due to the regular structures in our samples of man-made materials. We derived simple approximations for these reflectance characteristics using geometrical optics. PMID- 12645991 TI - Preformulation development of recombinant pegylated staphylokinase SY161 using statistical design. AB - The goal of this study was to perform preformulation development of SY161 by using statistical design methods to understand the effects of buffer strength, NaCl concentration, and pH on conformation and stability of the protein. It was also important to elucidate interactions between these factors. A central composite design using a 2-level full-factorial study was performed. Secondary structure was evaluated using circular dichroism. Stability toward unfolding was investigated using high-sensitivity differential scanning calorimetry. Depegylation, aggregation, and protein loss were evaluated using SEC-HPLC with on line light scattering, at time zero and after a 2-week stability study. Response surface plots clearly show optimal pH, NaCl, and buffer conditions. Interactions between pH and NaCl as well as pH and buffer concentration are observed. Tm is seen to be predictive of SY161 stability. Secondary structure changes were minimal and did not influence stability. Statistical design was very effective in providing an understanding of the effects of the formulation components on SY161 stability. PMID- 12645992 TI - Bioerodible injectable poly(ortho ester) for tetracycline controlled delivery to periodontal pockets: preliminary trial in humans. AB - The semisolid consistency of poly(ortho esters) (POEs) containing tetracycline free base allows direct injection in the periodontal pocket and shows sustained and almost constant in vitro release in phosphate buffer, pH 7.4 at 37 degrees C, for up to 14 days. Total polymer degradation concomitant with drug release was obtained. Formulations containing 10% or 20% (wt/wt) tetracycline were evaluated in a panel of 12 patients suffering from severe and recurrent periodontitis. In the first trial including 6 patients, single-rooted teeth and molar teeth with furcations were treated immediately after scaling and root planing. Patients tolerated both formulations well, experienced no pain during application, and showed no signs of irritation or discomfort during the observation period. However, retention of the formulation was minimal in this first study. An improved clinical protocol followed in the second study (stopping bleeding after scaling and root planning) prolonged the retention of the formulations in the inflamed periodontal pockets. For up to 11 days, tetracycline concentrations in the gingival crevicular fluid were higher than the minimum inhibitory concentration of tetracycline against most periodontal pathogens. PMID- 12645993 TI - Monitoring intracellular pH changes in response to osmotic stress and membrane transport activity using 5-chloromethylfluorescein. AB - Intracellular free H+ concentration (pHi) responds to numerous extracellular stimuli. The use of fluorescent indicator dyes to measure pHi is strongly influenced by the ability of target cells to retain activated dye within the cytoplasmic compartment. Here, 3 pH-sensitive indicator dye - acetoxymethyl (AM) esters of SNARF-1 and BCECF, and the thiol-reactive 5-chloromethyfluorescein (CMFDA) - were examined for monitoring pHi. The stability of pH measurements was strongly affected by temperature, cell type, indicator dye, and use of transport inhibitors to prevent dye export. Cellular retention of CMFDA, which forms covalent complexes, was sufficient to permit monitoring of transient pHi changes over extended time periods in a multi-well plate assay format. In human embryonic kidney (HEK293) and Chinese hamster ovary (CHO) cells, increasing osmotic pressure caused a significant rise in pHi. In contrast, activation of native or transfected beta-adrenergic, cholinergic, and d and m opioid receptors did not measurably affect pHi in HEK293 cells. Decreases in pHi were observed in CHO cells expressing the human H+/peptide transporter PEPT1 upon addition of dipeptide substrates. The use of CMFDA in multi-well formats should facilitate study of osmotic and transport activity and screening for drugs that affect pHi. PMID- 12645994 TI - Population analysis of the pharmacokinetics and pharmacodynamics of RWJ-270201 (BCX-1812) in treating experimental influenza A and B virus in healthy volunteers. AB - OBJECTIVE: Our objective was to assess the pharmacokinetics and pharmacodynamics of RWJ-270201 (BCX-1812), an oral neuraminidase inhibitor for the treatment of influenza A and B virus in healthy volunteers. METHODS: This was a double-blind, randomized, placebo-controlled, parallel group study. A total of 80 adult male and female subjects were enrolled for the influenza A challenge study. This was a 5-arm study (100 mg/qd, 200 mg/qd, 200 mg/bid, 400 mg/qd, and placebo). In the challenge B virus model, 60 subjects were enrolled for a 3-arm study (800 mg on Day 1 followed by 400 mg on Days 2-5; 800 mg on Days 1-5; and placebo). The pharmacokinetics of RWJ-270201 (BCX-1812) were characterized with the use of a population approach and were described by a 2-compartmental model with first order absorption and elimination. The pharmacodynamic data, mean log viral titers, were described with the use of an empirical equation relating the viral growth and the effect of drug on changes in viral titers. RESULTS: Pharmacokinetic analyses show that weight was the most significant covariate for all estimated pharmacokinetic parameters. The pharmacodynamic data, mean log viral titers showed a decrease in viral titers with increase in plasma exposure. The decrease in viral titer started to occur 12 hours following the drug dosing, and viral suppression lasted 72 hours to 96 hours. The exposures associated with a 50% decrease in viral titers were 1089 ng-h/mL and 1898 ng-h/mL, respectively. CONCLUSIONS: A PK/PD model was well utilized to characterize the effect of RWJ 270201 (BCX-1812) on the influenza A and B virus. The results from this model showed that both the loading dose and the standard dose regimens are efficacious against A and B virus. RWJ-270201 (BCX-1812) is under clinical development for the treatment of influenza A and B infections in adult and high-risk populations. It is a potent and selective inhibitor of both influenza A and B virus neuraminidases and inhibits the viral cleavage of sialic acid from cell surface glycoproteins and glycolipids. Consequently, RWJ-270201 (BCX-1812) prevents infection by stopping the release of newly formed virus from the surface of infected cells and preventing viral spread across the mucous lining of the respiratory tract. It therefore represents an attractive agent for antiviral therapy. PMID- 12645995 TI - Bioinformatic analysis of the human mu opioid receptor (OPRM1) splice and polymorphic variants. AB - Mu opioid receptor (OPRM1), a member of the G-protein coupled receptor superfamily, mediates the analgesic and euphoric effects of opioid drugs. The sequences of OPRM1 cDNA and reported splice variants were used to search the public and Celera genomic databases. The matched sequences were analyzed to assemble an OPRM1 genomic contig. Human OPRM1 gene was estimated to span at least 90 kb in the chromosome 6q24-25 region. Four coding exons are separated by 3 introns. While intron 2 has only 773 bp, these databases for the first time provide the precise length of and other information about long introns 1 and 3, containing 50 and 27 kb, respectively. When a consensus exon/intron splice junction at the end of the coding exon 3 was not utilized, it may have resulted in continuous translation of the exon to yield the splice variant OPRM1A. The study did not identify human orthologs of other OPRM1 variants that had been reported for mouse OPRM1, although several proposed exons were found to be included in mouse genomic clones. Single nucleotide polymorphisms in the OPRM1 gene were also analyzed and summarized, which could provide potential polymorphic markers for molecular genetic studies. PMID- 12645996 TI - Pharmacokinetics, mass balance, and tissue distribution of a novel DNA alkylating agent, VNP40101M, in rat. AB - VNP40101M (1,2-Bis(methylsulfonyl)-1-(2-chloroethyl)-2-[(2 methylamino)carbonyl] hydrazine), a novel DNA alkylating agent, is currently under clinical development for the treatment of cancer in Phase I clinical trials. This study investigated the pharmacokinetics, mass balance, and tissue distribution of [14C]-VNP40101M in rats following a single intravenous dose of 10 mg/kg. After 7 days, the total recovery of radioactivity was 85% for males and 79% for females. Most of the radioactivity was eliminated within 48 hours through urine (70%), with less excreted in feces (6%). Tissue contained relatively high radioactive residues with the highest concentrations in kidneys, liver, lung, and spleen. After 7 days, tissue still contained 9% of the dose. At both 5 minutes and 1 hour post dose, brain contained relatively high radioactivity (5.9 and 3.3 micro g equivalence/g and 50% and 30% of the blood concentration, respectively), suggesting that VNP40101M penetrated the blood-brain barrier. The elimination half-life of VNP40101M was approximately 20 minutes, the peak plasma concentration (Cmax) averaged 11.3 micro g/mL, the volume of distribution (Vss) averaged 0.91 L/kg, and the total body clearance (Cl) averaged 33.5 mL/min/kg. The metabolite profile in urine was complex, indicating VNP40101M was extensively metabolized. There were no apparent sex differences in pharmacokinetic parameters of VNP40101M in the rat. PMID- 12645997 TI - Which concentration of the inhibitor should be used to predict in vivo drug interactions from in vitro data? AB - When the metabolism of a drug is competitively or noncompetitively inhibited by another drug, the degree of in vivo interaction can be evaluated from the [I]u/Ki ratio, where [I]u is the unbound concentration around the enzyme and Ki is the inhibition constant of the inhibitor. In the present study, we evaluated the metabolic inhibition potential of drugs known to be inhibitors or substrates of cytochrome P450 by estimating their [I]u/Ki ratio using literature data. The maximum concentration of the inhibitor in the circulating blood ([I]max), its maximum unbound concentration in the circulating blood ([I]max,u), and its maximum unbound concentration at the inlet to the liver ([I]in,max,u) were used as [I]u, and the results were compared with each other. In order to calculate the [I]u/Ki ratios, the pharmacokinetic parameters of each drug were obtained from the literature, together with their reported Ki values determined in in vitro studies using human liver microsomes. For most of the drugs with a calculated [I]in,max,u/Ki ratio less than 0.25, which applied to about half of the drugs investigated, no in vivo interactions had been reported or "no interaction" was reported in clinical studies. In contrast, the [I]max,u/Ki and [I]max/Ki ratio was calculated to be less than 0.25 for about 90% and 65% of the drugs, respectively, and more than a 1.25-fold increase was reported in the area under the concentration-time curve of the co-administered drug for about 30% of such drugs. These findings indicate that the possibility of underestimation of in vivo interactions (possibility of false-negative prediction) is greater when [I]max,u or [I]max values are used compared with using [I]in,max,u values. PMID- 12645999 TI - Comparison of stepwise covariate model building strategies in population pharmacokinetic-pharmacodynamic analysis. AB - The aim of this study was to compare 2 stepwise covariate model-building strategies, frequently used in the analysis of pharmacokinetic-pharmacodynamic (PK-PD) data using nonlinear mixed-effects models, with respect to included covariates and predictive performance. In addition, the effects of stepwise regression on the estimated covariate coefficients were assessed. Using simulated and real PK data, covariate models were built applying (1) stepwise generalized additive models (GAM) for identifying potential covariates, followed by backward elimination in the computer program NONMEM, and (2) stepwise forward inclusion and backward elimination in NONMEM. Different versions of these procedures were tried (eg, treating different study occasions as separate individuals in the GAM, or fixing a part of the parameters when the NONMEM procedure was used). The final covariate models were compared, including their ability to predict a separate data set or their performance in cross-validation. The bias in the estimated coefficients (selection bias) was assessed. The model-building procedures performed similarly in the data sets explored. No major differences in the resulting covariate models were seen, and the predictive performances overlapped. Therefore, the choice of model-building procedure in these examples could be based on other aspects such as analyst- and computer-time efficiency. There was a tendency to selection bias in the estimates, although this was small relative to the overall variability in the estimates. The predictive performances of the stepwise models were also reasonably good. Thus, selection bias seems to be a minor problem in this typical PK covariate analysis. PMID- 12645998 TI - Peptide delivery to the brain via adsorptive-mediated endocytosis: advances with SynB vectors. AB - Biological membranes normally restrict the passage of hydrophilic molecules. This impairs the use of a wide variety of drugs for biomedical applications. To overcome this problem, researchers have developed strategies that involve conjugating the molecule of interest to one of a number of peptide entities that are efficiently transported across the cell membranes. In the past decade, a number of different peptide families with the ability to cross the cell membranes have been identified. Certain of these families enter the cells by a receptor independent mechanism, are short (10-27 amino acid residues), and can deliver successfully various cargoes across the cell membrane into the cytoplasm or nucleus. Surprisingly, some of these vectors, the SynB vectors, have also shown the ability to deliver hydrophilic molecules across the blood-brain barrier, one of the major obstacles to the development of drugs to combat diseases affecting the CNS. PMID- 12646000 TI - Effects of obesity on the pharmacodynamics of nitroglycerin in conscious rats. AB - Literature reports have suggested that hemodynamic response toward organic nitrates may be reduced in obese patients, but this effect has not been studied. We compared the mean arterial pressure (MAP) responses toward single doses of nitroglycerin (NTG, 0.5-50 micro g) in conscious Zucker obese (ZOB), Zucker lean (ZL), and Sprague-Dawley (SD) rats. NTG tolerance development in these animal groups was separately examined. Rats received 1 and 10 micro g/min of NTG or vehicle infusion, and the maximal MAP response to an hourly 30 micro g NTG IVchallenge dose (CD) was measured. Steady-state NTG plasma concentrations were measured during 10 micro g/min NTG infusion. The Emax and ED50 values obtained were 33.9 +/- 3.6 and 3.5 +/- 1.7 micro g for SD rats, 33.2 +/- 4.1 and 3.0 +/- 1.4 micro g for ZL rats, and 34.8 +/- 3.9 and 5.3 +/- 2.8 micro g for ZOB rats, respectively. No difference was found in the dose-response curves among these 3 groups (P >.05, 2-way ANOVA). Neither the dynamics of NTG tolerance development, nor the steady-state NTG plasma concentrations, were found to differ among these 3 animal groups. These results showed that ZOB rats are not more resistant to the hemodynamic effects of organic nitrates compared with their lean controls. Thus, the acute and chronic hemodynamic effects induced by NTG are not sensitively affected by the presence of obesity in a conscious animal model of genetic obesity. PMID- 12646001 TI - Detection of MDR1 single nucleotide polymorphisms C3435T and G2677T using real time polymerase chain reaction: MDR1 single nucleotide polymorphism genotyping assay. AB - The objective of this study was to develop a real-time polymerase chain reaction (PCR) method to detect MDR1 (human multidrug resistance gene) single nucleotide polymorphisms (SNPs) C3435T and G2677T. C3435T and G2677T are linked to MDR1*2, which is associated with enhanced efflux activity in vitro. Using the Smart Cycler, an allele-specific real-time PCR-based genotyping method was developed to detect C3435T and G2677T. The MDR1 genotype of human genomic DNA templates was determined by direct DNA sequencing. PCR reactions for genotyping C3435T and G2677T by using allele-specific primers were conducted in separate tubes. An additional nucleotide mismatch at the third position from the 3' end of each allele-specific primer was used to abrogate nonspecific PCR amplification. The fluorescence emitted by SYBR Green I was monitored to detect formation of specific PCR products. PCR growth curves exceeding the threshold cycle were considered positive. Fluorescence melt-curve analysis was used to corroborate results from PCR growth curves. Using PCR growth curves, our assay accurately determined hetero- and homozygosity for C3435T and G2677T. Genotype assignments based on PCR growth curve, melt-curve analysis, agarose gel electrophoresis, and direct DNA sequencing results of PCR products were in perfect agreement. We have developed a rapid MDR1 genotyping method that can be used to assess the contribution of MDR1*2 to pharmacokinetic and pharmacodynamic variability of P glycoprotein substrates. PMID- 12646002 TI - Role of plasma lipoproteins in modifying the toxic effects of water-insoluble drugs: studies with cyclosporine A. AB - Lipoproteins are a heterogeneous population of macromolecular aggregates of lipids and proteins that are responsible for the transport of lipids through the vascular and extravascular fluids from their site of synthesis or absorption to peripheral tissues. Lipoproteins are involved in other biological processes as well, including coagulation and tissue repair, and serve as carriers of a number of hydrophobic compounds within the systemic circulation. It has been well documented that disease states (eg, AIDS, diabetes, cancer) significantly influence circulating lipoprotein content and composition. Therefore, it appears possible that changes in the lipoprotein profile would affect not only the ability of a compound to associate with lipoproteins but also the distribution of the compound within the lipoprotein subclasses. Such an effect could alter the pharmacokinetics and pharmacological action of the drug. This paper reviews the factors that influence the interaction of one model hydrophobic compound, cyclosporine A, with lipoproteins and the implications of altered plasma lipoprotein concentrations on the pharmacological behavior of this compound. PMID- 12646004 TI - Linking human and veterinary health: trends, directions and initiatives. AB - The objective of this brief article is to provide an overview of some of the important harmonization efforts that are currently under way within the animal health community. Topics include: scientific networks and interdisciplinary communication; organizations that address animal-related public health concerns; the role of the veterinary pharmaceutical scientist within human health-oriented professional organizations; recent publications pertaining to veterinary pharmacology, pharmaceutics and therapeutics; and the role of global networking in veterinary product research and development. PMID- 12646003 TI - Quantitative comparison of functional screening by measuring intracellular Ca2+ with radioligand binding at recombinant human dopamine receptors. AB - The purpose of this study was to test whether screening at dopamine receptors performed with a recently described functional assay for G-protein coupled receptors (GPCRs) provides data that correlate significantly with radioligand binding data in the literature, thus possibly allowing researchers to replace radioligand binding with nonradioactive functional screening. Human dopamine receptors hD1 and hD2L (representing Gs [hD1] or Gi [hD2L] coupled GPCRs) were recombinantly expressed in human embryonic kidney (HEK293) cells. Cells were loaded with Oregon Green 488 BAPTA-1/AM and evenly distributed in 384 well plates. Seventeen test compounds were screened for agonistic activity by injection into the cell suspension and monitoringH of intracellular Ca2+ with a fluorescence microplate reader. Then, standard agonists (100nM SKF38393 for hD1, 30nM quinpirole for hD2L) were injected into wells preincubated with test compounds (screening for antagonism). Injection of various agonists resulted in a concentration-dependent increase in fluorescence. Further, preincubation of antagonists with dopamine receptor expressing cells inhibits concentration dependent the agonist-induced increase in fluorescence. Calculated apparent functional Ki values correlate with radioligand binding data in the literature (r2 = 0.7796 for D1, r2 = 0.7743 for D2). The correlation between apparent functional Ki values and radioligand binding data for the 17 tested compounds suggests that screening of test compounds at dopamine receptors with the functional Ca2+ assay can replace radioligand binding studies. Furthermore, besides apparent Ki values, information about agonistic or antagonistic properties of a test compound can be obtained with the functional Ca2+ assay. PMID- 12646005 TI - Evolving importance of biologics and novel delivery systems in the face of microbial resistance. AB - Methods to control infectious diseases in livestock are growing in importance. As the size of the average farm increases - for poultry, dairy and beef cattle, swine, and fish - the risk of rapid spread of infectious diseases increases as well. This increases the need for alternative methods of control of infectious agents. Improvements in specific immunogens, adjuvants, and delivery systems are needed to meet the demand for vaccines to ensure a healthy and safe meat supply. This article explores the challenges, trends, and recent advances in the control of infectious diseases through the use of biologics. PMID- 12646007 TI - Dissolution test development for complex veterinary dosage forms: oral boluses. AB - Fundamental aspects of electrolyte chemistry were used to design an appropriate dissolution medium with the capacity to maintain sink conditions throughout the test. Dissolution of various bolus dosage forms was studied using USP Apparatus II at various stirring speeds. Complete dissolution of each drug in the designed medium was achieved, and there is evidence that such a dissolution test could be discriminating. This review details the development of potentially discriminating in vitro dissolution tests for veterinary boluses using USP Apparatus II and examines the potential role of such testing during product quality assessments, in the evaluation of postapproval manufacturing changes and for the establishment of the generic equivalence of veterinary products. PMID- 12646006 TI - Interspecies considerations in the evaluation of human food safety for veterinary drugs. AB - Residues are composed of the parent drug and metabolites, and therefore interspecies comparisons must involve a consideration of comparative xenobiotic metabolism. The focus of this article will be the residue studies that are required to establish human food safety, and the interspecies pharmacokinetic differences and similarities that impact drug residues in animal- derived foods. To illustrate the factors that can complicate and assist these comparisons, 2 drugs will be examined in detail: ivermectin and fenbendazole. In addition, the activities of 2 US programs, the Food Animal Residue Avoidance Databank (FARAD) and the NRSP-7 (National Research Support Project Number 7) Minor Use Animal Drug Program will be presented, along with strategies that may be employed in the study of species differences. PMID- 12646008 TI - The role of the clinical pharmacologist in animal health. AB - Like most scientific disciplines, pharmacology is replete with subspecialties. Certainly most scientists recognize the value of animal studies in drug development for human pharmaceuticals. However, animals as the target species also represent a major focus of investigation. According to recent estimates, in the United States for the year 2000, 98.1 million cattle, 59.8 million pigs, and 1.5 billion chickens existed. Added to that estimate were companion animals, including 4 million horses, 59 million cats, and 52.9 million dogs. The estimate does not include the so-called "minor" species, such as 7 million sheep and 320,000 acres of freshwater fish production. In most respects, the medical needs of these animals are addressed in a manner parallel to that of human medicine. One such parallel, with certain distinct differences from its human counterpart, is veterinary clinical pharmacology. PMID- 12646009 TI - Impact of antimicrobial resistance on regulatory policies in veterinary medicine: status report. AB - Increasing resistance to antimicrobial agents is of growing concern to public health officials worldwide. The concern includes infections acquired in hospitals, community infections acquired in outpatient care settings, and resistant foodborne disease associated with drug use in food-producing animals. In the United States, a significant source of antimicrobial-resistant foodborne infections in humans is the acquisition of resistant bacteria originating from animals. The US Food and Drug Administration's (FDA's) goal in resolving the public health impact arising from the use of antimicrobial drugs in food producing animals is to ensure that significant human antimicrobial therapies are not compromised or lost while providing for the safe use of antimicrobials in food animals. The FDA's approach to the problem is multipronged and innovative. The strategy includes revision of the pre-approval safety assessment for new animal drug applications, use of risk assessment to determine the human health effect resulting from the use of antimicrobials in food animals, robust monitoring for changes in susceptibilities among foodborne pathogens to drugs that are important both in human and veterinary medicine, research, and risk management. PMID- 12646010 TI - Pharmacokinetic/pharmacodynamic integration in drug development and dosage regimen optimization for veterinary medicine. AB - Pharmacokinetic (PK)/pharmacodynamic (PD) modeling is a scientific tool to help developers select a rational dosage regimen for confirmatory clinical testing. This article describes some of the limitations associated with traditional dose titration designs (parallel and crossover designs) for determining an appropriate dosage regimen. It also explains how a PK/PD model integrates the PK model (describing the relationship between dose, systemic drug concentrations, and time) with the PD model (describing the relationship between systemic drug concentration and the effect vs time profile) and a statistical model (particularly, the intra- and interindividual variability of PK and/or PD origin). Of equal importance is the utility of these models for promoting rational drug selection on the basis of effectiveness and selectivity. PK/PD modeling can be executed using various approaches, such as direct versus indirect response models and parametric versus nonparametric models. PK/PD concepts can be applied to individual dose optimization. Examples of the application of PK/PD approaches in veterinary drug development are provided, with particular emphasis given to nonsteroidal anti-inflammatory drugs. The limits of PK/PD approaches include the development of appropriate models, the validity of surrogate endpoints, and the acceptance of these models in a regulatory environment. PMID- 12646012 TI - Deposition of nanoparticles in the arterial vessel by porous balloon catheters: localization by confocal laser scanning microscopy and transmission electron microscopy. AB - Restenosis remains the major limitation of percutaneous transluminal angioplasty (PTA) and stenting in the treatment of patients with atherosclerotic disease. Catheter-based local delivery of pharmacologic agents offers a potential therapeutic approach to reducing restenosis and minimizing undesirable systemic side effects. However, the intramural retention of liquid agents is low. Therefore, to achieve a sustained and regional release of the therapeutic agent it must be encapsulated in nanoparticle carrier systems. The purpose of this study was to investigate the size dependence of the penetration of nanoparticles after local delivery into the vessel wall of the aorta abdominalis of New Zealand white rabbits. Two milliliters of a 0.025% fluorescence-labeled polystyrene nanoparticle suspension with diameters ranging from 110 to 514 nm were infused at 2 atm and at constant PTA pressure of 8 atm into the aorta abdominalis. After the infused segments were removed, the location of nanoparticles was visualized using confocal laser scanning microscopy and transmission electron microscopy. The study demonstrates a size-dependent nanoparticle penetration into the intact vessel wall. While nanoparticles of about 100 and 200 nm were deposited in the inner regions of the vessel wall, 514-nm nanoparticles accumulated primarily at the luminal surface of the aorta. The observations confirm that size plays a critical role in the distribution of particles in the arterial vessel wall. It is additionally influenced by the formation of pressure-induced infusion channels, as well as by the existence of anatomic barriers, such as plaques, at the luminal surface of the aorta or the connective elastic tissue. PMID- 12646011 TI - An in vitro examination of the impact of polyethylene glycol 400, Pluronic P85, and vitamin E d-alpha-tocopheryl polyethylene glycol 1000 succinate on P glycoprotein efflux and enterocyte-based metabolism in excised rat intestine. AB - The potential inhibitory effects of 3 excipients (polyethylene glycol [PEG] 400, Pluronic P85, and vitamin E d-a-tocopheryl polyethylene glycol 1000 succinate [TPGS]) on the P-glycoprotein (P-gp) -mediated efflux of digoxin (DIG) and cytochrome P450 3A (CYP3A) -mediated metabolism of verapamil (VRP) have been examined in an in vitro permeability model. Experiments were conducted utilizing rat jejunal tissue mounted in diffusion chambers and included assessment of the serosal to mucosal (s to m) transport of DIG and the formation of norverapamil (NOR) during the mucosal to serosal transport of VRP, as measures of P-gp efflux and CYP3A metabolism, respectively. The presence of PEG at 1%, 5%, and 20% (wt/vol) reduced both the s to m flux of DIG (by 47%, 57%, and 64%, respectively, when compared to control) and the metabolism of VRP (by 54%, 78%, and 100%) in a concentration-dependent manner. P85 (0.1% wt/vol) significantly reduced s to m DIG flux by 47% and inhibited VRP metabolism by 42%. TPGS had insignificant effects on both metabolism and efflux at a concentration of 0.01% (wt/vol). The P gp inhibitory effects of PEG and P85 were evident regardless of whether the excipient was added to the mucosal side, the serosal side, or both sides of the tissue. The current data suggest that inclusion of PEG and P85 as solubilizing agents during in vitro permeability assessment may have a significant impact on both drug metabolism and efflux processes. These compounds appear to exert their effects on P-gp primarily via direct transporter inhibition - or indirectly, through effects on buffer osmolarity, membrane fluidity, and/or mitochondrial toxicity and subsequent adenosine triphosphate (ATP) depletion. PMID- 12646013 TI - Pharmacodynamic modeling of chemotherapeutic effects: application of a transit compartment model to characterize methotrexate effects in vitro. AB - The time course of chemotherapeutic effect is often delayed relative to the time course of chemotherapeutic exposure. In many cases, this delay is difficult to characterize mathematically through the use of standard pharmacodynamic models. In the present work, we investigated the relationship between methotrexate (MTX) exposure and the time course of MTX effects on tumor cell growth in culture. Two cancer cell lines, Ehrlich ascites cells and sarcoma 180 cells, were exposed for 24 hours to MTX concentrations that varied more than 700-fold (0.19-140 micro g/mL). Viable cells were counted on days 1, 3, 5, 7, 9, 11, 13, 15, 17, 20, 22, and 24 for Ehrlich ascites cells and on days 1, 2, 3, 5, 7, 9, 11, 13, 14, 15, 17, 19, and 21 for sarcoma 180 cells, through the use of a tetrazolium assay. Although MTX was removed 24 hours after application, cell numbers reached nadir values more than 100 hours after MTX exposure. Data from each cell line were fitted to 3 pharmacodynamic models of chemotherapeutic cell killing: a cell cycle phase-specific model, a phase-nonspecific model, and a transit compartment model (based on the general model recently reported by Mager and Jusko, Clin Pharmacol Ther. 70:210-216, 2001). The transit compartment model captured the data much more accurately than the standard pharmacodynamic models, with correlation coefficients ranging from 0.86 to 0.999. This report shows the successful application of a transit compartment model for characterization of the complex time course of chemotherapeutic effects; such models may be very useful in the development of optimization strategies for cancer chemotherapy. PMID- 12646015 TI - A thermogravimetric analysis of non-polymeric pharmaceutical plasticizers: kinetic analysis, method validation, and thermal stability evaluation. AB - Four non-polymeric plasticizers, propylene glycol, diethyl phthalate, triacetin, and glycerin have been subjected to rising temperature thermogravimetry for kinetic analysis and vaporization-based thermal stability evaluation. Since volatile loss of a substance is a function of its vapor pressure, the thermal stability of these plasticizers has been analyzed by generating vapor pressure curves using the Antoine and Langmuir equations. Unknown Antoine constants for the sample compounds, triacetin and glycerin have been derived by subjecting the vapor pressure curves to nonlinear regression. For the first time, the entire process of obtaining the unknown Antoine constants through thermogravimetry has been validated by developing an approach called the 'double reference method.' Based on this method, it has been possible to show that this technique is accurate even for structurally diverse compounds. Kinetic analysis on the volatilization of compounds revealed a predominant zero order process. The activation energy values for vaporization of propylene glycol, diethyl phthalate, triacetin, and glycerin, as deduced from the Arrhenius plots, have been determined to be 55.80, 66.45, 65.12, and 67.54 kJ/mol, respectively. The enthalpies of vaporization of the compounds have been determined from the Clausius-Clapeyron plots. Rising temperature thermogravimetry coupled with nonlinear regression analysis has been shown to be an effective and rapid technique for accurately predicting the vapor pressure behavior and thermal stability evaluation of volatile compounds. PMID- 12646014 TI - Chitosan and sodium alginate-based bioadhesive vaginal tablets. AB - Metronidazole was formulated in mucoadhesive vaginal tablets by directly compressing the natural cationic polymer chitosan, loosely cross-linked with glutaraldehyde, together with sodium alginate with or without microcrystalline cellulose (MCC). Sodium carboxymethylcellulose (CMC) was added to some of the formulations. The drug content in tablets was 20%. Drug dissolution rate studies from tablets were carried out in buffer pH 4.8 and distilled water. Swelling indices and adhesion forces were also measured for all formulations. The formula (FIII) containing 6% chitosan, 24% sodium alginate, 30% sodium CMC, and 20% MCC showed adequate release properties in both media and gave lower values of swelling index compared with the other examined formulations. FIII also proved to have good adhesion properties with minimum applied weights. Moreover, its release properties (% dissolution efficiency, DE) in buffer pH 4.8, as well as release mechanism (n values), were negligibly affected by aging. Thus, this formula may be considered a good candidate for vaginal mucoadhesive dosage forms. PMID- 12646016 TI - The prediction of plasma and brain levels of 2,3,5,6-tetramethylpyrazine following transdermal application. AB - The purpose of this study was to construct a pharmacokinetic (PK) model and to determine PK parameters of 2,3,5,6-tetramethylpyrazine (TMP) after application of TMP transdermal delivery system. Data were obtained in Sprague-Dawley (SD) rats following a single dose of TMP transdermal delivery system. Blood samples were obtained at 0, 0.25, 0.5, 1, 2, 4, 6, 16, and 24 hours after the transdermal application. In the brain level study, 18 SD rats were divided into 6 groups. Three SD rats before and after transdermal application were culled and sacrificed at each of the following time intervals: 2, 4, 6, 16, and 24 hours after the TMP TTS application. TMP concentrations in plasma and brain tissues were determined using high performance liquid chromatography and data were fitted using a zero order absorption and a first-order-elimination 3-compartment PK model. Fitted parameters included 2 volumes of distribution (V1, V2) and 2 elimination rate constants (k10, k20). The elimination half-life for TMP in plasma and brain was 26.5 and 31.2 minutes, respectively. The proposed PK model fit observed concentrations of TMP very well. This model is useful for predicting drug concentrations in plasma and brain and for assisting in the development of transdermal systems. PMID- 12646017 TI - Antiestrogens and selective estrogen receptor modulators as multifunctional medicines. 2. Clinical considerations and new agents. PMID- 12646018 TI - Synthesis and biological evaluation of non-peptidic cyclophilin ligands. AB - Peptidylprolyl isomerase cyclophilins play critical roles in a variety of biological processes. Recent findings that cyclophilins are present at high levels in the CNS and that cyclosporin A may possess neuroprotective/neurotrophic effects have prompted us to search for nonimmunosuppressant small molecule cyclophilin ligands. To this end, we report the lead identification through "virtual screening" and the synthesis of our first series of non-peptidic cyclophilin ligands, along with the preliminary biological results. PMID- 12646020 TI - Pyrrolopyrazinedione-based inhibitors of human hormone-sensitive lipase. AB - The regulation of lipid metabolism and it's effect on glucose control and diabetes has received intense interest. Hormone-sensitive lipase (HSL) is a vital enzyme in lipid metabolism. A series of novel pyrrolopyrazinediones has been discovered that demonstrate submicromolar activity both in the enzyme assay and in a (14)C-emulsion assay employing cholesteryl oleate as a substrate as a secondary measure of HSL activity. These compounds represent novel inhibitors of the human HSL enzyme. PMID- 12646019 TI - Discovery of 5-[5-fluoro-2-oxo-1,2- dihydroindol-(3Z)-ylidenemethyl]-2,4- dimethyl-1H-pyrrole-3-carboxylic acid (2-diethylaminoethyl)amide, a novel tyrosine kinase inhibitor targeting vascular endothelial and platelet-derived growth factor receptor tyrosine kinase. AB - To improve the antitumor properties and optimize the pharmaceutical properties including solubility and protein binding of indolin-2-ones, a number of different basic and weakly basic analogues were designed and synthesized. 5-[5-Fluoro-2-oxo 1,2-dihydroindol-(3Z)-ylidenemethyl]-2,4-dimethyl-1H-pyrrole-3-carboxylic acid (2 diethylaminoethyl)amide (12b or SU11248) has been found to show the best overall profile in terms of potency for the VEGF-R2 and PDGF-Rbeta tyrosine kinase at biochemical and cellular levels, solubility, protein binding, and bioavailability. 12b is currently in phase I clinical trials for the treatment of cancers. PMID- 12646021 TI - Discovery of tyrosine-based potent and selective melanocortin-1 receptor small molecule agonists with anti-inflammatory properties. AB - The melanocortin-1 receptor (MC-1R) is a G-protein-coupled receptor involved in inflammation and skin pigmentation. Compound 2 is the first highly potent and selective MC-1R small-molecule agonist reported. Compound 2 showed efficacy in an acute model of inflammation, which has demonstrated the role of MC-1R in modulation of inflammation. PMID- 12646022 TI - Design and synthesis of naphthalenic dimers as selective MT1 melatoninergic ligands. AB - We report the synthesis and binding properties at MT(1) and MT(2) receptors of the first example of agomelatine (N-[2-(7-methoxynaphth-1-yl)ethyl]acetamide) dimers in which two agomelatine moieties are linked together through their methoxy substituent by a polymethylene side chain according to the "bivalent ligand" approach. Some of these compounds behave as MT(1)-selective ligands. The most selective one (5) behaves as an antagonist. PMID- 12646023 TI - In vitro and in vivo antitumor properties of tetrakis((trishydroxy- methyl)phosphine)gold(I) chloride. AB - A novel hydrophilic gold compound, tetrakis((trishydroxymethyl)phosphine)gold(I) chloride 1, has been investigated for its antitumor properties. In vitro studies demonstrate that 1 is active against HCT-15, AGS, PC-3, and LNCaP tumor cells. Cell cycle analysis of the HCT-15 cells by flow cytometry revealed elongation of the G1 phase of the cell cycle leading to growth inhibition. Administration of 1 to Balb/C mice inoculated with syngenic meth/A cells demonstrated statistically significant dose-dependent survival time. PMID- 12646024 TI - Virtual screening for submicromolar leads of tRNA-guanine transglycosylase based on a new unexpected binding mode detected by crystal structure analysis. AB - Eubacterial tRNA-guanine transglycosylase (TGT) is involved in the hypermodification of cognate tRNAs, leading to the exchange of G34 by preQ1 at the wobble position in the anticodon loop. Mutation of the tgt gene in Shigella flexneri results in a significant loss of pathogenicity of the bacterium due to inefficient translation of a virulence protein mRNA. Herein, we describe the discovery of a ligand with an unexpected binding mode. On the basis of this binding mode, three slightly deviating pharmacophore hypotheses have been derived. Virtual screening based on this composite pharmacophore model retrieved a set of potential TGT inhibitors belonging to several compound classes. All nine tested inhibitors being representatives of these classes showed activity in the micromolar range, two of them even in the submicromolar range. PMID- 12646025 TI - Design, synthesis, and pharmacological profile of novel fused pyrazolo[4,3 d]pyridine and pyrazolo[3,4-b][1,8]naphthyridine isosteres: a new class of potent and selective acetylcholinesterase inhibitors. AB - A new family of tacrine (THA) analogues (7-9, 12), containing the azaheterocyclic pyrazolo[4,3-d]pyridine or pyrazolo[3,4-b][1,8]naphthyridine systems as isosteres of the quinoline ring of THA, has been synthesized. The compounds were tested in rat brain cholinesterases using Ellman's method, and all were fully efficacious in inhibiting the enzymes. Compounds 9 and 12b were the most potent against acetylcholinesterase (AChE), showing IC(50) of 6.0 and 6.4 microM, and were less active against rat brain butyrylcholinesterase, showing selectivity indexes of 5.3 and 20.9, respectively. Compounds 7-9 and 12 were also tested for their acute neurotoxicity in vitro, using cultured rat cortical cells. Compounds 7 and 8 were not significantly toxic; 9 was toxic at 500 microM, but not at 100 microM. The naphthyridine derivatives 12a and 12b showed a significant concentration dependent neurotoxicity, being able to kill most cells at 500 microM. Molecular dynamic simulation using the X-ray crystal structure of AChE from Torpedo californica was used to explain the possible binding mode of these new THA isosteres. PMID- 12646026 TI - Use of a pharmacophore model to discover a new class of influenza endonuclease inhibitors. AB - Data from both our own and literature studies of the biochemistry and inhibition of influenza virus endonuclease was combined with data on the mechanism of action and the likely active site mechanism to propose a pharmacophore. The pharmacophore was used to design a novel structural class of inhibitors, some of which were found to have activities similar to that of known influenza endonuclease inhibitors and were also antiviral in cell culture. PMID- 12646027 TI - Synthesis of novel thrombin inhibitors. Use of ring-closing metathesis reactions for synthesis of P2 cyclopentene- and cyclohexenedicarboxylic acid derivatives. AB - The thrombin inhibitory tripeptide d-Phe-Pro-Arg has been mimicked using either cyclopentenedicarboxylic derivatives or a cyclohexenedicarboxylic derivative as surrogate for the P2 proline. In the P3 position, tertiary amides were optimized as d-Phe P3 replacements. The P1 arginine was, in all compounds, substituted with the more rigid and biocompatible 4-aminomethylbenzamidine. One of the novel inhibitors was cocrystallized with alpha-thrombin and subjected to X-ray analysis. From analysis of the X-ray crystal structure, new ligands were designed leading to significantly improved binding affinity, the lead candidate exhibiting an in vitro IC(50) of 49 nM. PMID- 12646028 TI - Modes of methyleneoxy bridging and their effect on tetrahydronaphthalene lignan cytotoxicity. AB - Dioxatricyclodecane, oxabicyclooctane, and benzodihydropyran derivatives of alpha conidendrin (ACON), podophyllotoxin (PT), and sikkimotoxin (SK) were prepared to learn which methyleneoxy bridging modes and arene and aryl substituents coincided with high cytotoxicity. PT-derived dioxatricyclodecane 14 showed in vitro activity at 10(-8) M. SK analogue 12 was less active, and ACON analogue 11 was inactive at 10(-4) M. In vivo intraperitoneal and subcutaneous activities of 14 were observed. In vitro cytotoxicities were higher for oxabicyclooctanes when hydroxymethyl group and methyleneoxy bridge were cis, as in deoxypicropodophyllin analog20, rather than trans, as in PT analogue 5. Acetylation of the hydroxymethyl group of 20 lowered activities, whereas acetylation of 5 increased or lowered activities. Reduction of the hydroxymethyl group of 5 to a methyl group increased cytotoxicities. Molecular dynamics indicated the THN scaffold of benzodihydropyrans was conformationally mobile, but scaffolds of oxabicyclooctanes and dioxatricyclodecanes were immobile. Each of three PT benzodihydropyrans was less active than its oxabicyclooctane counterpart. PMID- 12646029 TI - New active series of growth hormone secretagogues. AB - New growth hormone secretagogue (GHS) analogues were synthesized and evaluated for growth hormone releasing activity. This series derived from EP-51389 is based on a gem-diamino structure. Compounds that exhibited higher in vivo GH-releasing potency than hexarelin in rat (subcutaneous administration) were then tested per os in beagle dogs and for their binding affinity to human pituitary GHS receptors and to hGHS-R 1a. Compound 7 (JMV 1843, H-Aib-(d)-Trp-(d)-gTrp-formyl) showed high potency in these tests and was selected for clinical studies.(1) PMID- 12646030 TI - Role of the glycopeptide framework in the antibacterial activity of hydrophobic derivatives of glycopeptide antibiotics. AB - The antibacterial properties of glycopeptide antibiotics are based on their interaction with the d-Ala-d-Ala containing pentapeptide of bacterial peptidoglycan. The hydrophobic amides of vancomycin (1), teicoplanin (2), teicoplanin aglycon (3), and eremomycin (4) were compared with similar amides of minimally or low active des-(N-methyl-d-leucyl)eremomycin (5), eremomycin aglycon (6), des-(N-methyl-d-leucyl)eremomycin aglycon (7), and a teicoplanin degradation product TB-TPA (8). All hydrophobic amides of 1, 3, 4, and 6 were almost equally active against glycopeptide-resistant enterococci (GRE) [minimum inhibitory concentrations (MIC) 150 mm) morphology. Spherical, tubular, and filamentous conceptuses were collected from pregnant gilts and subjected to SSH. Forward and reverse subtractions were performed to identify candidate genes differentially expressed during spherical to tubular and tubular to filamentous transition. A total of 384 transcripts were differentially screened to ensure unique expression. Of the transcripts screened, sequences were obtained for 142 that were confirmed to be differentially expressed among the various morphologies. Gene expression profiles during rapid trophoblastic elongation were generated for selected mRNAs using quantitative real-time PCR. During the transition from tubular to early filamentous conceptuses, s-adenosylhomocysteine hydrolase and heat shock cognate 70 kDa expression were significantly enhanced. A novel unknown gene was isolated and shown to be significantly up-regulated at the onset of rapid trophoblastic elongation and further enhanced in filamentous conceptuses. PMID- 12646054 TI - Expression of the insulin-like growth factor (IGF) system and steroidogenic enzymes in canine testis tumors. AB - Testis tumors occur frequently in dogs. The main types of tumors are Sertoli cell tumors, seminomas, and Leydig cell tumors. Mixed tumors and bilateral occurrence of tumors may be encountered frequently. To elucidate the possible relationship between the insulin-like growth factor (IGF) system and the development of different types of testis tumors in dogs, the expression of insulin-like growth factor-I and II (IGF-I and IGF-II), their type I receptor (IGF-IR), and their binding proteins (IGFBPs) was examined. In addition the expression of the steroidogenic enzymes p450-aromatase and 5alpha-reductase type I and type II, and the androgen receptor (AR) was investigated by a semiquantitative reverse transcriptase PCR (RT-PCR). Both normal testes and testes with tumors were studied. In normal testes a clear expression of IGF-I, IGF-II, IGF-IR, IGFBP2, IGFBP4 and IGFBP5 was found. Expression of IGFBP1 and IGFBP3 was weak. There was also clear expression of the steroidogenic enzymes 5alpha-reductase, aromatase, and the AR. Quantification of RT-PCR products revealed significantly less expression of IGFBP1, IGF-I, and 5alpha-reductase type I in Sertoli cell tumors and seminomas. Leydig cell tumors and mixed tumors had a significantly higher expression of IGFBP4 and IGF-IR than normal testes. The expression of aromatase was lower in seminomas and in mixed tumors. The expression of AR, IGF-II and IGFBP2, IGFBP3, IGFBP5, and 5alpha-reductase type II did not differ among the different types of tumors. It was concluded that Sertoli cell tumors and seminomas have a comparable expression of the IGF system while Leydig cell tumors have a different pattern, suggesting difference in pathobiology among these types of tumors. PMID- 12646055 TI - Male pheromone stimulates ovarian follicular development and body growth in juvenile female opossums (Monodelphis domestica). AB - Female opossums are induced into estrus by direct exposure to a non-volatile pheromone in male scentmarks. Juvenile females develop this responsiveness by 150 days of age (days), and earlier (130 days) if exposed to male pheromone beginning at 90 days. The goal of this study was to examine the effect of male pheromone on body growth and ovarian follicular development in young opossums. Females (n = 28) were weaned at 56 days and caged individually with ad libitum food and water from 84 days. Body mass was recorded 2-3 times weekly and the onset of estrus was monitored by urogenital sinus cytology. Exposed females were given continuous access to adult male scentmarks from 90 days and randomly selected for necropsy at 105 and 130 days. Unexposed females were isolated from direct contact with males and their scentmarks and randomly selected for necropsy at 90, 105, 130, and 150 days. Exposed females were larger (63.5 +/- 1.1 g) than unexposed females (56.6 +/- 1.1 g) at 130 days, and 4 of 5 had expressed estrus or proestrus. Uterine mass at 130 days was higher (P < 0.05) in exposed (129.8 +/- 28.8 mg) than in unexposed (25.4 +/- 6.7 mg) females, none of which expressed estrus by 150 days. The mean number of developing, antral follicles per female increased from 1.5 +/- 1.5 at 90 days to 17.8 +/- 5.2 at 130 days. Mean diameter of developing antral follicles at 130 days was larger in exposed (534 +/- 54 microm) than in unexposed females (393 +/- 4 microm). The results of this study demonstrate that pheromonal induction of first estrus in juvenile opossums is associated with an increased rate of body growth and follicular development. PMID- 12646058 TI - Expression of somatostatin receptor type-2 (sst2A) in immature porcine Leydig cells and a possible role in the local control of testosterone secretion. AB - We recently reported that immature porcine Leydig cells express both somatostatin (SRIF) and SRIF receptor type-2 (sst-2) transcripts. The present study was therefore undertaken to assess whether SRIF might exert autocrine actions on these cells through sst2A receptor, one of the two sst2 isoforms known to exert important neuroendocrine and endocrine functions. Using a polyclonal antibody directed towards the C-terminal tail of the sst2A receptor subtype, receptor immunoreactivity was detected in a subpopulation of Leydig cells and spermatogonia. To address the physiological correlates of this expression we then studied the possible involvement of sst2 receptor in the regulation of testosterone secretion. Functional assays showed that the sst2 agonist octreotide inhibited both basal and hCG-stimulated testosterone secretion by testosterone pretreated Leydig cells. To assess whether sst2 receptor expression might be regulated by testosterone, we performed a semi-quantitative RT-PCR analysis of sst2 mRNA expression in Leydig cells cultured in the presence or in the absence of the androgen. A significant increase in sst2 receptor transcripts was observed in testosterone-treated cells. Taken together, these data suggest that SRIF can inhibit testosterone secretion through the sst2A receptor. The mechanism of the local inhibitory actions of SRIF is probably autocrine since immature porcine Leydig cells express SRIF itself and it might involve testosterone-induced increase of sst2 receptor expression in immature Leydig cells. PMID- 12646056 TI - Proposed mechanism for sperm chromatin condensation/decondensation in the male rat. AB - Condensation of sperm chromatin occurs after spermatozoa have left the caput epididymis and are in transit to the cauda epididymis, during which time large numbers of disulfide bonds are formed. The formation of these disulfide bonds requires the repeated oxidation of the cofactor, NAD(P)H. To date, the means by which this oxidation is achieved has yet to be elucidated. Spermatozoa lose the bulk of their cytoplasm prior to leaving the testis; and, as a result, any shuttle systems for removing and transferring reducing equivalents into the mitochondria are unlikely to be operational. In an apparent preparation for the loss of cytoplasm, however, the following events occur during spermatogenesis. First, androgen-binding protein (ABP) is produced by the Sertoli cells of the testis; second, high affinity binding sites for ABP are inserted into the membrane surrounding the nucleus; and third, a nuclear location is acquired for the enzyme, 3alpha-hydroxysteroid dehydrogenase (3alpha-HSD). We propose that after the loss of cytoplasm, the nuclear region of spermatozoa is directly accessible to constituents contained in the lumen of the caput epididymis. As a consequence, luminal ABP attaches itself to the nuclear membrane via its binding sites, and is internalized. After internalization, ABP exerts its principle function, which is to bind to luminal 5alpha-dihydrotestosterone (5alpha-DHT), thereby ensuring its availability to the enzyme, 3alpha-HSD. In the conversion of 5alpha-DHT to 3alpha-androstanediol (3alpha-Diol), NAD(P)H is oxidized. Spermatozoa that reach the cauda epididymis have fully condensed chromatin. In addition, the nuclear region retains appreciable amounts of 5alpha-DHT and 3alpha Diol, both bound to ABP. During fertilization, the bound 3alpha-Diol is converted back to 5alpha-DHT, reducing equivalents are transferred to NAD(P)+, and disulfide bonds are broken.IVF clinics report that spermatozoa with incompletely condensed chromatin have a low percentage of fertilization. If our proposed mechanism for chromatin condensation/decondensation is borne out by further research, IVF clinics might consider preincubating spermatozoa with 5alpha-DHT in order to increase the efficiency of fertilization. PMID- 12646057 TI - Transmembrane and truncated (SEC) isoforms of MUC1 in the human endometrium and Fallopian tube. AB - The cell surface mucin MUC1 is expressed by endometrial epithelial cells with increased abundance in the secretory phase of the menstrual cycle, when it is found both at the apical cell surface and in secretions. This suggests the presence of a maternal cell surface glycoprotein barrier to embryo implantation, arising from the anti-adhesive property of MUC1. In previous work, we demonstrated alternatively spliced MUC1 variant forms in tumour cells. The variant MUC1/SEC lacks the transmembrane and cytoplasmic sequences found in the full-length variant. We now show that MUC1/SEC mRNA is present in endometrial carcinoma cell lines, endometrial tissue and primary cultured endometrial epithelial cells. The protein can be detected using isoform-specific antibodies in uterine flushings, suggesting release from endometrium in vivo. However, on the basis of immunolocalisation studies, MUC1/SEC also remains associated with the apical epithelial surface both in tissue and in cultured cells. Transmembrane MUC1 and MUC1/SEC are both strikingly localised to the apical surface of tubal epithelium. Thus MUC1 may contribute to the anti-adhesive character of the tubal surface, inhibiting ectopic implantation. The mechanism by which this barrier is overcome in endometrium at implantation is the subject of ongoing investigation. PMID- 12646059 TI - Signaling mechanisms in tumor necrosis factor alpha-induced death of microvascular endothelial cells of the corpus luteum. AB - The microvasculature of the corpus luteum (CL), which comprises greater than 50% of the total number of cells in the CL, is thought to be the first structure to undergo degeneration via apoptosis during luteolysis. These studies compared the apoptotic potential of various cytokines (tumor necrosis factor alpha, TNFalpha; interferon gamma, IFNgamma; soluble Fas ligand, sFasL), a FAS activating antibody (FasAb), and the luteolytic hormone prostaglandin F2alpha (PGF2alpha) on CL derived endothelial (CLENDO) cells. Neither sFasL, FasAb nor PGF2alpha had any effect on CLENDO cell viability. Utilizing morphological and biochemical parameters it was evident that TNFalpha and IFNgamma initiated apoptosis in long term cultures. However, TNFalpha was the most potent stimulus for CLENDO cell apoptosis at early time points. Unlike many other studies described in non reproductive cell types, TNFalpha induced apoptosis of CLENDO cells occurs in the absence of inhibitors of protein synthesis. TNFalpha-induced death is typically associated with acute activation of distinct intracellular signaling pathways (e.g. MAPK and sphingomyelin pathways). Treatment with TNFalpha for 5-30 min activated MAPKs (ERK, p38, and JNK), and increased ceramide accumulation. Ceramide, a product of sphingomyelin hydrolysis, can serve as an upstream activator of members of the MAPK family independently in numerous cell types, and is a well-established pro-apoptotic second messenger. Like TNFalpha, treatment of CLENDO cells with exogenous ceramide significantly induced endothelial apoptosis. Ceramide also activated the JNK pathway, but had no effect on ERK and p38 MAPKs. Pretreatment of CLENDO cells with glutathione (GSH), an intracellular reducing agent and known inhibitor of reactive oxygen species (ROS) or TNFalpha-induced apoptosis, significantly attenuated TNFalpha-induced apoptosis. It is hypothesized that TNFalpha kills CLENDO cells through elevation of reactive oxygen species, and intracellular signals that promote apoptosis. PMID- 12646060 TI - Synchronous onset of oestradiol-17beta secretion by Meishan conceptuses. AB - The response of Meishan conceptuses to an exogenous precursor for oestradiol 17beta biosynthesis was investigated in vitro, to determine whether gestational age or morphological stage of development elicit changes in hormone metabolism. Conceptuses were recovered on days 11, 12, 13 or 15 after the onset of oestrus and cultured for 6 hours at 37 degrees C, in the presence or absence of testosterone. On days 12 and 13 after the onset of oestrus spherical conceptuses were recovered from some gilts, whereas others yielded elongated or filamentous conceptuses. All conceptuses recovered on day 15 after oestrus had elongated. The number of cells per individual conceptus increased from days 11 to 13 after the onset of oestrus (P < 0.001), as did conceptus surface area (P = 0.038). Supplementing culture media with testosterone, as a substrate for oestrogen biosynthesis, significantly increased conceptus oestradiol-17beta secretion in vitro on days 12, 13 and 15, regardless of whether pre- or post-elongation conceptuses were cultured. However, on day 11 oestradiol-17beta was only detected at significant concentrations in the culture media of four testosterone supplemented conceptuses and only one gilt produced conceptuses capable of secreting oestradiol-17beta in the absence of testosterone. Therefore, the onset of conceptus oestradiol-17beta secretion is apparently limited by the expression of aromatase enzymes that are activated synchronously, irrespective of the stage of morphological development, within Meishan litters. Once established, Meishan conceptus oestradiol-17beta secretion in vitro is increased in the presence of exogenous testosterone. PMID- 12646061 TI - Responsiveness of bovine cumulus-oocyte-complexes (COC) to porcine and recombinant human FSH, and the effect of COC quality on gonadotropin receptor and Cx43 marker gene mRNAs during maturation in vitro. AB - Substantially less development to the blastocyst stage occurs in vitro than in vivo and this may be due to deficiencies in oocyte competence. Although a large proportion of bovine oocytes undergo spontaneous nuclear maturation, less is known about requirements for proper cytoplasmic maturation. Commonly, supraphysiological concentrations of FSH and LH are added to maturation media to improve cumulus expansion, fertilization and embryonic development. Therefore, various concentrations of porcine FSH (pFSH) and recombinant human FSH (rhFSH) were investigated for their effect on bovine cumulus expansion in vitro. Expression of FSHr, LHr and Cx43 mRNAs was determined in cumulus-oocyte complexes to determine whether they would be useful markers of oocyte competence. In serum free media, only 1000 ng/ml pFSH induced marked cumulus expansion, but the effect of 100 ng/ml pFSH was amplified in the presence of 10% serum. In contrast, cumulus expansion occurred with 1 ng/ml rhFSH in the absence of serum. FSHr mRNA was highest at 0-6 h of maturation, then abundance decreased. Similarly, Cx43 mRNA expression was highest from 0-6 h but decreased by 24 h of maturation. However, the relative abundance of LHr mRNA did not change from 6-24 h of maturation. Decreased levels of FSHr, LHr and Cx43 mRNAs were detected in COCs of poorer quality. In conclusion, expansion of bovine cumulus occurred at low doses of rhFSH in serum-free media. In summary, FSHr, LHr and Cx43 mRNA abundance reflects COC quality and FSHr and Cx43 mRNA expression changes during in vitro maturation; these genes may be useful markers of oocyte developmental competence. PMID- 12646062 TI - Expression and localization of estrogen receptor-alpha protein in normal and abnormal term placentae and stimulation of trophoblast differentiation by estradiol. AB - Estrogens play an important role in the regulation of placental function, and 17 beta-estradiol (E2) production rises eighty fold during human pregnancy. Although term placenta has been found to specifically bind estrogens, cellular localization of estrogen receptor alpha (ER-alpha) in trophoblast remains unclear. We used western blot analysis and immunohistochemistry with h-151 and ID5 monoclonal antibodies to determine the expression and cellular localization of ER-alpha protein in human placentae and cultured trophoblast cells. Western blot analysis revealed a ~65 kDa ER-alpha band in MCF-7 breast carcinoma cells (positive control). A similar band was detected in five normal term placentae exhibiting strong expression of Thy-1 differentiation protein in the villous core. However, five other term placentae, which exhibited low or no Thy-1 expression (abnormal placentae), exhibited virtually no ER-alpha expression. In normal placentae, nuclear ER-alpha expression was confined to villous cytotrophoblast cells (CT), but syncytiotrophoblast (ST) and extravillous trophoblast cells were unstained. In abnormal placentae no CT expressing ER-alpha were detected. Normal and abnormal placentae also showed ER-alpha expression in villous vascular pericytes and amniotic (but not villous) fibroblasts; no staining was detected in amniotic epithelial cells or decidual cells. All cultured trophoblast cells derived from the same normal and abnormal placentae showed distinct ER-alpha expression in western blots, and the ER-alpha expression was confined to the differentiating CT, but not to the mature ST. Trophoblast cells from six additional placentae were cultured in normal medium with phenol red (a weak estrogen) as above (PhR+), or plated in phenol red-free medium (PhR-) without or with mid-pregnancy levels of E2 (20 nM). Culture in PhR- medium without E2 caused retardation of syncytium formation and PhR-medium with E2 caused acceleration of syncytium formation compared to cultures in PhR+ medium. These data indicate that the considerable increase in estrogen production during pregnancy may play a role, via the ER-alpha, in the stimulation of CT differentiation and promote function in normal placentae. This mechanism, however, may not operate in abnormal placentae, which show a lack of ER-alpha expression. PMID- 12646064 TI - Ovarian follicle counts--not as simple as 1, 2, 3. PMID- 12646063 TI - Effects of deletion of the prolactin receptor on ovarian gene expression. AB - Prolactin (PRL) exerts pleiotropic physiological effects in various cells and tissues, and is mainly considered as a regulator of reproduction and cell growth. Null mutation of the PRL receptor (R) gene leads to female sterility due to a complete failure of embryo implantation. Pre-implantatory egg development, implantation and decidualization in the mouse appear to be dependent on ovarian rather than uterine PRLR expression, since progesterone replacement permits the rescue of normal implantation and early pregnancy. To better understand PRL receptor deficiency, we analyzed in detail ovarian and corpora lutea development of PRLR-/- females. The present study demonstrates that the ovulation rate is not different between PRLR+/+ and PRLR-/- mice. The corpus luteum is formed but an elevated level of apoptosis and extensive inhibition of angiogenesis occur during the luteal transition in the absence of prolactin signaling. These modifications lead to the decrease of LH receptor expression and consequently to a loss of the enzymatic cascades necessary to produce adequate levels of progesterone which are required for the maintenance of pregnancy. PMID- 12646066 TI - Peptides identify multiple hotspots within the ligand binding domain of the TNF receptor 2. AB - BACKGROUND: Hotspots are defined as the minimal functional domains involved in protein:protein interactions and sufficient to induce a biological response. RESULTS: Here we describe the use of complex and high diversity phage display libraries to isolate peptides (called Hotspot Ligands or HSPLs) which sub-divide the ligand binding domain of the tumor necrosis factor receptor 2 (TNFR2; p75) into multiple hotspots. We have shown that these libraries could generate HSPLs which not only subdivide hotspots on protein and non-protein targets but act as agonists or antagonists. Using this approach, we generated peptides which were specific for human TNFR2, could be competed by the natural ligands, TNFalpha and TNFbeta and induced an unexpected biological response in a TNFR2-specific manner. CONCLUSIONS: To our knowledge, this is the first report describing the dissection of the TNFR2 into biologically active hotspots with the concomitant identification of a novel and unexpected biological activity. PMID- 12646067 TI - mRNA analysis of single living cells. AB - Analysis of specific gene expression in single living cells may become an important technique for cell biology. So far, no method has been available to detect mRNA in living cells without killing or destroying them. We have developed here a novel method to examine gene expression of living cells using an atomic force microscope (AFM). AFM tip was inserted into living cells to extract mRNAs. The obtained mRNAs were analyzed with RT-PCR, nested PCR, and quantitative PCR. This method enabled us to examine time-dependent gene expression of single living cells without serious damage to the cells. PMID- 12646068 TI - Self-assembly of proteins and their nucleic acids. AB - We have developed an artificial protein scaffold, herewith called a protein vector, which allows linking of an in-vitro synthesised protein to the nucleic acid which encodes it through the process of self-assembly. This protein vector enables the direct physical linkage between a functional protein and its genetic code. The principle is demonstrated using a streptavidin-based protein vector (SAPV) as both a nucleic acid binding pocket and a protein display system. We have shown that functional proteins or protein domains can be produced in vitro and physically linked to their DNA in a single enzymatic reaction. Such self assembled protein-DNA complexes can be used for protein cloning, the cloning of protein affinity reagents or for the production of proteins which self-assemble on a variety of solid supports. Self-assembly can be utilised for making libraries of protein-DNA complexes or for labelling the protein part of such a complex to a high specific activity by labelling the nucleic acid associated with the protein. In summary, self-assembly offers an opportunity to quickly generate cheap protein affinity reagents, which can also be efficiently labelled, for use in traditional affinity assays or for protein arrays instead of conventional antibodies. PMID- 12646069 TI - BioMagnetic Research and Technology: a new online journal. PMID- 12646071 TI - The utilisation of health research in policy-making: concepts, examples and methods of assessment. AB - The importance of health research utilisation in policy-making, and of understanding the mechanisms involved, is increasingly recognised. Recent reports calling for more resources to improve health in developing countries, and global pressures for accountability, draw greater attention to research-informed policy making. Key utilisation issues have been described for at least twenty years, but the growing focus on health research systems creates additional dimensions.The utilisation of health research in policy-making should contribute to policies that may eventually lead to desired outcomes, including health gains. In this article, exploration of these issues is combined with a review of various forms of policy-making. When this is linked to analysis of different types of health research, it assists in building a comprehensive account of the diverse meanings of research utilisation.Previous studies report methods and conceptual frameworks that have been applied, if with varying degrees of success, to record utilisation in policy-making. These studies reveal various examples of research impact within a general picture of underutilisation.Factors potentially enhancing utilisation can be identified by exploration of: priority setting; activities of the health research system at the interface between research and policy-making; and the role of the recipients, or 'receptors', of health research. An interfaces and receptors model provides a framework for analysis.Recommendations about possible methods for assessing health research utilisation follow identification of the purposes of such assessments. Our conclusion is that research utilisation can be better understood, and enhanced, by developing assessment methods informed by conceptual analysis and review of previous studies. PMID- 12646070 TI - Breast cancer screening beliefs by practice location. AB - BACKGROUND: This study examines variations in breast cancer screening among primary care clinicians by geographic location of clinical practice. METHODS: A cross-sectional survey design was used to examine approaches to breast cancer screening among physicians, nurse practitioners, and physician assistants involved in primary care practice. A summary index of beliefs about breast cancer screening was created by summing the total number of responses in agreement with each of four survey items; values for this summary variable ranged between zero and four. Respondents were classified into urban, rural and suburban categories based upon practise location. RESULTS: Among the 428 respondents, agreement with "correct" responses ranged from 50% to 71% for the individual survey items; overall, half agreed with three or more of the four breast cancer screening items. While no significant differences were noted by practice location, variation in responses were evident. Reported use of written breast cancer guidelines was less in both suburban (OR = 0.51) and urban areas (OR = 0.56) when compared to clinicians in rural areas. CONCLUSION: Development of an evidence based consensus statement regarding breast cancer screening would support a single set of unambiguous guidelines for implementation in all primary care settings, thus decreasing variations in how breast cancer screening is approached across varied clinical settings. PMID- 12646072 TI - Assessing capacity for health policy and systems research in low and middle income countries* AB - BACKGROUND: As demand grows for health policies based on evidence, questions exist as to the capacity of developing countries to produce the health policy and systems research (HPSR) required to meet this challenge. METHODS: A postal/web survey of 176 HPSR producer institutions in developing countries assessed institutional structure, capacity, critical mass, knowledge production processes and stakeholder engagement. Data were projected to an estimated population of 649 institutions. RESULTS: HPSR producers are mostly small public institutions/units with an average of 3 projects, 8 researchers and a project portfolio worth $155,226. Experience, attainment of critical mass and stakeholder engagement are low, with only 19% of researchers at PhD level, although researchers in key disciplines are well represented and better qualified. Research capacity and funding are similar across income regions, although inequalities are apparent. Only 7% of projects are funded at $100,000 or more, but they account for 54% of total funding. International sources and national governments account for 69% and 26% of direct project funding, respectively. A large proportion of international funds available for HPSR in support of developing countries are either not spent or spent through developed country institutions. CONCLUSIONS: HPSR producers need to increase their capacity and critical mass to engage effectively in policy development and to absorb a larger volume of resources. The relationship between funding and critical mass needs further research to identify the best funding support, incentives and capacity strengthening approaches. Support should be provided to network institutions, concentrate resources and to attract funding. PMID- 12646073 TI - Apamin inhibits NO-induced relaxation of the spontaneous contractile activity of the myometrium from non-pregnant women. AB - There is now considerable evidence for the involvement of K+ channels in nitric oxide (NO) induced relaxation of smooth muscles including the myometrium. In order to assess whether apamin-sensitive K+ channels play a role in NO - induced relaxation of the human uterus, we have studied the effect of specific blockers of these channels on the relaxation of myometrium from non-pregnant women. In vitro isometric contractions were recorded in uterine tissues from non-pregnant premenopausal women who had undergone hysterectomy. Apamin (10 nM) and scyllatoxin (10 nM) did not alter spontaneous myometrial contractions. However, 15-min pretreatment of the myometrium strips with apamin completely inhibited relaxation caused by diethylamine-nitric oxide (DEA/NO). The pretreatment with scyllatoxin significantly reduced (about 2.6 times) maximum relaxation of the strips induced by DEA/NO (p < 0.05). These results strongly suggest that, beside Ca2+ and voltage dependent charybdotoxin-sensitive (CTX-sensitive) K+ channels, apamin-sensitive K+ channels are also present in the human non-pregnant myometrium. These channels offer an additional target in the development of new tocolytic agents. PMID- 12646074 TI - Effect of long-term treatment with steroid hormones or tamoxifen on the progesterone receptor and androgen receptor in the endometrium of ovariectomized cynomolgus macaques. AB - The progesterone receptor (PR) and androgen receptor (AR) belong to the nuclear receptor superfamily. Two isoforms of PR (A and B) have been identified with different functions. The expression of AR, each isoform of PR and their involvement in long-term effects on the endometrium after hormonal replacement therapy (HRT) or tamoxifen (TAM) treatment is not known. The aims of this study were to determine PR(A+B), PRB and AR distribution by immunohistochemistry in the macaque (Macaca fascicularis) endometrium. Ovariectomized (OVX) animals were orally treated continuously for 35 months with either conjugated equine estrogens (CEE); medroxyprogesterone acetate (MPA); the combination of CEE/MPA; or TAM. Treatment with CEE/MPA tended to down-regulate PR in the superficial glands, but increased it in the stroma. TAM treatment increased both the PR and PRB levels in the stroma. Overall, less than 20% of the cells were positive for the PRB isoform and less variation was observed after steroid treatment. AR was found in the stroma, mainly distributed in the basal layer of the endometrium in the OVX and steroid treated groups, but was absent in the TAM treated group. No AR was found in the glandular epithelium. The present data show that long-term hormone treatment affects the PR level, and also the ratio between PRA and PRB in the endometrium. PMID- 12646075 TI - Ovarian responses to undernutrition in pregnant ewes, USA. AB - In most mammals oogonia proliferate by mitosis and begin meiotic development during fetal life. Previous studies indicated that there is a delay in the progression to the first stage of meiotic arrest in germ cells of female fetuses of undernourished ewes. We report that underfeeding (50% NRC requirement beginning on Day 28 of pregnancy) provokes an increase in oxidative base lesions within DNA of mid-gestational (Day 78) fetal oogonia; this condition was associated with up-regulation of the tumor suppressor/cell-cycle arrest modulator p53, antiapoptotic factor Bcl-2, and base-excision repair polymerase beta. Fetal ovarian weights and germ cell concentrations were not altered by nutrient deprivation. Ovaries of ewes on control diets (100% NRC) contained more tertiary follicles than their restricted counterparts; however, peripheral venous estradiol-17beta was not different between groups. There was no effect of treatment on p53 accumulation in maternal oocytes. Luteal structure-function was not perturbed by undernutrition. No fetal losses were attributed to the dietary restriction. It is proposed that DNA of interphase fetal oogonia is vulnerable to oxidative insults perpetrated by a nutritional stress to the dam, and that multiple/integrated adaptive molecular response mechanisms of cell-cycle inhibition (providing the time required for base repairs) and survival hence sustain the genomic integrity and population stability of the germline. PMID- 12646078 TI - [Calcitonin today]. PMID- 12646076 TI - Relaxin receptors and nitric oxide synthases: search for the missing link. PMID- 12646079 TI - [Estimating the costs associated with nosocomial urinary tract infection. A case control study]. AB - BACKGROUND: Nosocomial infection (NI) is associated with increased resource use. The current study addressed the cost excess attributable to nosocomial urinary tract infection (NUTI). METHODS: Case-control study conducted in a cohort population. For matching, the Diagnostic Related Group (DRG) to which the episode of case patient was ascribed was used. Then, a further search was conducted in the hospital discharge database with the following parameters: DRG, gender, age, admission date, department, comparison of length of stays, main diagnosis, co morbidities, number of secondary diagnoses and procedures. Matching was achieved for 64 episodes (71%), and upon them an estimation of costs was performed. RESULTS: The mean length of stay for cases were 15.3 (median: 12) and 12.3 (median: 11) days for cases and controls, respectively (p = 0.0001). The excess of length of stay attributable to NUTI was 3 days (95% CI 1.6-4.7), longer for patients admitted to Medical Departments (5.3 days) than for patients admitted to Surgical Departments (2 days) (p = 0,03). The use of diagnostic resources was significantly higher for bacteriological testing only. The use of antibiotics and fluid therapy was higher among infected patients. Out of the total excess of the estimated costs per episode, 132,047 ptas, 93% corresponded to the increase in hospital stay. Ten patients (15.6%) were responsible for 68% of the total of extra-costs. In 17 occasions (26.6%), the control patient used more resources than the infected patient. CONCLUSIONS: Nosocomial urinary tract infection is associated with a resource use directly related to its presence. For the most part, it is related to the prolongation of hospital stay. PMID- 12646077 TI - Differentiation of adult-type Leydig cells occurs in gonadotrophin-deficient mice. AB - During mammalian testis development distinct generations of fetal and adult Leydig cells arise. Luteinising hormone (LH) is required for normal adult Leydig cell function and for the establishment of normal adult Leydig cell number but its role in the process of adult Leydig cell differentiation has remained uncertain. In this study we have examined adult Leydig cell differentiation in gonadotrophin-releasing hormone (GnRH)-null mice which are deficient in circulating gonadotrophins. Adult Leydig cell differentiation was assessed by measuring expression of mRNA species encoding four specific markers of adult Leydig cell differentiation in the mouse. Each of these markers (3beta hydroxysteroid dehydrogenase type VI (3betaHSD VI), 17beta-hydroxysteroid dehydrogenase type III (17betaHSD III), prostaglandin D (PGD)-synthetase and oestrogen sulphotransferase (EST)) is expressed only in the adult Leydig cell lineage in the normal adult animal. Real-time PCR studies showed that all four markers are expressed in adult GnRH-null mice. Localisation of 3betaHSD VI and PGD-synthetase expression by in situ hybridisation confirmed that these genes are expressed in the interstitial tissue of the GnRH-null mouse. Treatment of animals with human chorionic gonadotrophin increased expression of 3betaHSD VI and 17betaHSD III within 12 hours further indicating that differentiated, but unstimulated cells already exist in the GnRH-null mouse. Thus, while previous studies have shown that LH is required for adult Leydig cell proliferation and activity, results from the present study show that adult Leydig cell differentiation will take place in animals deficient in LH. PMID- 12646080 TI - [Is the routine chest radiography following thoracentesis necessary?]. AB - OBJECTIVES: To assess the routine indication of chest roentgenogram after thoracentesis in function of iatrogenic pneumothorax causation, following a procedure protocol. PATIENTS AND METHODS: Prospective study of 104 consecutive thoracentesis procedures performed in 76 patients. The procedure was protocolized and after each thoracentesis a chest roentgenogram was obtained within 6 hours after the procedure. The attending physician completed a questionnaire relative to patients data characteristics of the pleural effusion. RESULTS: Two pneumothorax were noted following thoracentesis (2%) which did not require pleural drainage. These pneumothorax occurred in patients with parapneumonic pleural effusions. CONCLUSIONS: The routine practice of chest roentgenogram after thoracentesis does not seem to be warranted and should be individualized. The performance of thoracentesis by physicians in training period, with procedure protocolization, minimizes the risk of pneumothorax. PMID- 12646081 TI - [Effect of lipoprotein (a) on the risk of coronary heart disease in adults aged less than 65 years in function of gender differences]. AB - BACKGROUND: Lipoprotein (a) is an independent risk factor for coronary heart disease, particularly the premature form. However, differences in its effect have been observed between men and women and in different populations. SUBJECTS AND METHODS: Lipid and non-lipid risk factors were compared (univariate and multivariate analyses) between a group of patients with myocardial infarction aged less than 65 years (n = 33) and those in their siblings (n = 32) and an unrelated healthy control group (n = 45) of the same gender. RESULTS: No differences were observed between male patients and their siblings. Differences were indeed observed when patients when compared with controls regarding HDL cholesterol (p = 0.001) and lipoprotein(a) (p = 0.002) serum concentrations. Female patients also had lower serum concentrations of HDL cholesterol than their siblings and controls, but non significant differences were observed for lipoprotein (a). The analysis of lipoprotein (a), categorized with respect to the 90th percentile of controls for each gender, showed a significant association with coronary heart disease only for males: crude odds ratio (OR) compared with controls 10.7 (95% CI: 2.1-54.9) and adjusted of 9.9 (95% CI: 1.6-60.7). The strength of this association was smaller and non-significant in women: crude OR 6,4 (95% CI: 0.8-51.8) and adjusted of 3.09 (95% CI: 0.3-47.2). CONCLUSIONS: Lipoprotein (a) concentrations are similar in siblings and increased concentrations in our population are more strongly and clearly association with coronary heart disease in men. PMID- 12646083 TI - [Current indications for proton pump inhibitors]. PMID- 12646082 TI - [Patient orientation clinic: an initiative to reduce waiting lists]. AB - OBJECTIVE: The patient orientation clinic (POC), acting as a filter, is an initiative aimed at reducing the waiting lists for medical specialities. METHODS: For a six-months period, an internist and a registered nurse attended 1,265 patients proposed for those specialties. RESULTS: 85.7% of cases were referred to their general practitioners after evaluation at our clinic. The remaining patients (14.3%) went on and frecuently benefited from initial therapies or because complementary tests and/or examinations were arranged for and earlier date. CONCLUSIONS: POC is shaping as a promising tool for health resource management in reducing waiting lists in medical specialties. PMID- 12646084 TI - [Computed tomography in the diagnosis of tuberculous peritonitis]. PMID- 12646085 TI - [Update in the diagnosis and differential diagnosis of Cushing's syndrome]. PMID- 12646086 TI - [Bilateral massive pneumothorax]. PMID- 12646087 TI - [Extreme weakness associated with gastric adenocarcinoma: beyond the expected]. PMID- 12646088 TI - [Eruption-like presentation of erythematous-desquamative lesions]. PMID- 12646089 TI - [Primary amyloidosis associated with osteosclerotic myeloma]. PMID- 12646090 TI - [Hypercholesterolemia, rhabdomyolysis and hypothyroidism]. PMID- 12646091 TI - [Inclusion body myositis with respiratory onset]. PMID- 12646092 TI - [Persistent fever, pancytopenia and spleen enlargement in a heart transplant carrier as presentation of visceral leishmaniasis]. PMID- 12646093 TI - [Applications of somatostatin receptor scintigraphy in oncology: the need to incorporate SPECT into the imaging protocol]. PMID- 12646095 TI - [Results of treatment with calculated doses of radioiodine depending on gland size and morphology in graves' disease. Analysis of variables]. AB - OBJECTIVE: To assess the efficacy of the administered doses of Radioiodine (RAI) in the treatment of patients with hyperthyroidism in Graves' disease, calculated on the basis of gland size and morphology (assessment by palpation and scintigraphy), analyzing which variables influence the final result of our sample, and on the early development of hypothyroidism. METHODS: 99 patients treated with RAI between april 1996 and november 2000 were evaluated retrospectively, 77 women and 22 men, mean age 44 years (range: 18-76). 67 patients received treatment with oral antithyroid drugs. The administered dose range was: 185-629 MBq. Mean follow-up time was 13.4 months. Success was defined as elimination of hyperthyroidism; and early hypothyroidism which appeared in the six months after therapy. RESULTS: 30 patients remained with euthyroidism, 53 patients had hypothyroidism and 16 had recurrent hyperthyroidism. A total of 84% of the patients were controlled with one dose of RAI. Only the time of evolution influenced the final results. The rest of the variables (gender, age, doses and antithyroid drugs) did not influence them. No relationship was found between the appearance of early/late hypothyroidism and the administered doses. A certain tendency was found for early hypothyroidism to appear in the youngest patients (p = 0.111). CONCLUSIONS: The administered doses in our department have been shown to be effective in controlling thyrotoxicosis. In our sample, only time of evolution influenced on the final results of the patients. Only the age factor showed a certain tendency in the evolution to early/late hypothyroidism. PMID- 12646094 TI - [Somatostatin receptor scintigraphy. When would a SPECT study be performed?]. AB - Somatostatin receptor scintigraphy has become an essential tool in the treatment planning of patients with carcinoid tumor (CaT) and pancreatic neuroendocrine tumors (PNET). However, the contribution of SPECT images in the diagnosis and staging of these tumors has not been well established. OBJECTIVE: 1) To find out if SPECT improves the sensitivity of planar imaging and adds any value to tumor diagnosis, and 2) To determine under which circumstances SPECT images must be obtained. MATERIAL AND METHODS: We have evaluated 49 patients, 26 females and 23 males, using both planar and SPECT 111In-DTPAOC images. The patients were classified into three groups according to their clinical indication: Group A: Indolent symptoms of CaT or PNET (n = 9); Group B: Known or functioning CaT or PNET tumors (n = 22); Group C: Post-surgical follow-up (n = 18). The total number of SPECT studies were 51 (37 abdomen, 13 thorax and 1 brain). In all cases SPECT was performed 24 hours after 111In-DTPAOC injection. Scintigraphic images were compared with surgical findings in 17 patients and with a synthesis of biochemical, clinical and CT findings in the other patients. RESULTS: 16 primary lesions were detected with planar images whereas SPECT detected 18. The two tumors detected only by SPECT were a bronchial CaT and an insulinoma located in the pancreatic tail. Planar and SPECT image sensitivities were 66.6% and 75% respectively. Twenty one patients developed metastatic disease. Sensitivities of planar and SPECT images for metastases were 76.1% and 90.4% respectively. Furthermore, SPECT helps planar images in the evaluation of six patients with uncertain images (lesions with low receptor density or with an uncertain scintigraphic localization). Therefore, in 15.6% of patients with primary or metastatic tumors, planar images were normal whereas SPECT was abnormal and SPECT provided an added value in 21.8% of the studies with abnormal planar images. CONCLUSIONS: We recommend the use of SPECT images in: 1) Patients with specific functional clinical syndrome or with a known CaT or PNET and normal planar images. 2) When planar images are abnormal only in primary tumor. 3) In order to confirm the absence of other metastases in patients with known liver metastases. 4) In order to enhance the detection of lesions with low receptor density and uncertain planar images and 5) In order to determine exactly the anatomic location of some lesions PMID- 12646096 TI - [CD44s Cell surface expression in 55 lung adenocarcinomas. Negative correlation with tumor size and CA125 cytosolic levels]. AB - INTRODUCTION: CD44s belongs to a family of cell adhesion molecules involved in cell adhesion, migration and cell-extracellular matrix interactions. In this work we attempt to study CD44s expression in lung adenocarcinomas and its possible correlation with other clinicobiological parameters. MATERIAL AND METHODS: Using an EIA, cell surface CD44s levels were determined in 55 lung adenocarcinomas, classified according to clinical stage, histological grade, ploidy and cellular S phase fraction. CA125 cytosolic concentrations were also assayed. RESULTS: Forty two adenocarcinomas (76.4%) showed CD44s concentrations > 80 ng/mg prot, and did not differ significantly from those observed in 16 normal samples (93.7%). There were no differences in CD44s expression when clinical stage (I: 24/28, II: 6/9 and III: 12/17), lymph node involvement (N = 245/29, N+: 18/26), ploidy (diploid: 3/5, aneuploid: 32/39), histological grade (I: 6/7, III: 18/26) and cellular S phase (> 8.8%: 24/31, < or = 8.8%: 17/24) were considered. Positive CD44s tumors had lower CA125 (p: 0.0072) cytosolic levels and a reduced tumor size (p: 0.0093). CONCLUSIONS: CD44s expressions in lung adenocarcinomas did not correlate with any clinicobiological parameters, but there was a negative correlation between this and reduced tumor size and lower CA125 cytosolic levels. PMID- 12646097 TI - [S100 protein as tumoral marker in melanoma patients. Comparative study with sentinel node biopsy and whole body FDG-PET]. AB - PURPOSE: Analyze the significance of serum S100 as tumor marker and study it in the sentinel node biopsy and in the whole body FDG-PET studies. MATERIAL AND METHODS: 332 melanoma patients diagnosed of cutaneous melanoma were included (41 untreated and 291 previously treated). Serum levels were measured by an immunoradiometric assay. Cut-off value was 0.2 microg/l. RESULTS: In the 41 untreated patients, the S100 mean values were: mean 0.36 microg/l, median 0.29 microg/l, with 56.1% of positivities. Serum mean values in stage III and IV were significantly higher than stage I and II (0.489 microg/l vs 0.209 microg/l) (p < 0.05). The mean serum value of S100 in treated patients and without evidence of disease was 0.14 microg/l and the median 0.13 microg/l. Sensitivity and specificity for detection of recurrence or metastases in the follow-up were 76.2% and 88.6%, respectively. There is a correlation of 91.3% between S100 and FDG-PET studies (23 patients). S100 does not provide information on the sentinel lymph node biopsy. CONCLUSIONS: S100 is not useful to diagnose melanoma in its early stages but it is a good tumor marker for the follow-up and monitoring of melanoma patients. PMID- 12646099 TI - [Detection of superior vena cava stenosis with superior vena cava-superior left pulmonary artery shunt and pulmonary hypertension. Detection by nuclear medicine study]. AB - We present a 59 year old patient diagnosed of right superior lobe pulmonary carcinoma by CT scan. A ventilation/perfusion pulmonary scintigraphy quantified for study of pulmonary function after surgery was performed. The images show an abnormal pattern of the distribution of macroaggregate albumin (MAA) and extrapulmonary presence of tracer (right/left shunt). A first step dynamic study showed the existence of superior cava stenosis and passage of the tracer from the vena cava to the left pulmonary artery by collateral circulation (innominate vein) that not only explained the existence of the abnormal pattern of the MAA distribution but also what the cause of the right/left shunt was. After the resolution of the stenosis, the distribution pattern became normal and the extrapulmonary presence of MAA disappeared. PMID- 12646098 TI - [Bilateral vocal cords paresis following iodine therapy]. AB - We present a case of a 33 year old man who underwent a thyroidectomy due to papillary thyroid carcinoma, with a reversible paresis of both vocal cords after radioiodine therapy. The patient had no previous lesion in laryngeal nerves. Paralysis of the vocal cords is a rare complication of the 131I administration and generally occurs more frequently when there is already some type of lesion in the recurrent laryngeal nerves. The paper reviews the literature on this type of complications. PMID- 12646100 TI - [Bilateral agenesis of parotid glands: scintigraphic demonstration]. PMID- 12646101 TI - [Enteric salmonella pancarditis. Diagnosis of site by examination with 67Gallium]. PMID- 12646102 TI - [Finding a thymoma in a 99mTc-Tetrofosmin myocardial perfusion imaging]. PMID- 12646103 TI - [Osteoid osteoma diagnosed by three phase bone scintigraphy in a young woman with back pain]. PMID- 12646104 TI - [PET and SPECT of cerebral neurotransmission]. PMID- 12646105 TI - [Biological features characteristic of the mucinous carcinoma of the breast]. PMID- 12646107 TI - [Prevalence of postpartum depression in Spanish mothers: comparison of estimation by mean of the structured clinical interview for DSM-IV with the Edinburgh Postnatal Depression Scale]. AB - BACKGROUND AND OBJECTIVE: Our objectives were: To estimate the prevalence of postpartum depression (PD) by two evaluation methods: Structured Clinical Interview for DSM-IV (SCID), and self-report Edinburgh Postnatal Depression Scale (EPDS), and to identify the EPDS cut-off to provide an unbiased estimation of PD prevalence. PATIENTS AND METHOD: We report data from all women (n = 1191) attending to postnatal routine checking visits at six weeks postpartum during one year in the Gynaecology and Obstetrics Department of the Hospital Clinic of Barcelona (Spain). A two-stage method was used. In the first stage, all women selected completed the EPDS. In the second stage, all women with an EPDS score >= 9 (probable PD cases) and a randomized sample of 16% with a score < 9 were evaluated by a psychiatrist using the SCID interview to establish a psychiatric diagnosis of major and minor depression. 402 postpartum women were offered the SCID interview; 68 of them refused to participate. RESULTS: The prevalence of depression according to the SCID interview was 10.15% (CI 95%, 8.43-11.87). The prevalence of major depression was 3.6% (CI 95%, 2.55-4.67) and it was 6.5% (CI 95%, 5.14-7.95) for minor depression. An EPDS cut-off of 11/12 provided an unbiased estimation of the postpartum depression prevalence rate. CONCLUSIONS: Our results justify the need to use different EPDS cut-offs. A cut-off of 10/11 is effective for identifying the population at risk and a cut-off of 11/12 is useful to estimate the prevalence in epidemiological studies. PMID- 12646106 TI - [A prospective, controlled, non-randomized study of three cohorts to test the effectiveness of two antibiotics levofloxacin and cotrimoxazol on antibacterial prophylaxis in neutropenic patients]. AB - BACKGROUND AND OBJECTIVE: At present, there is strong concern about the efficacy of current antimicrobial prophylaxis for the management of neutropenic patients. The purpose of this study was to test the effectiveness of levofloxacin, a new quinolone with expanded activity against grampositive bacteria, versus cotrimoxazol as a prophylactic treatment for granulocytopenic patients. PATIENTS AND METHOD: In this prospective and controlled study, we included 249 consecutive episodes of neutropenia, such as those resulting from lymphoma and leukemia treatment, during 28 months (from November 1999 to February 2002). These episodes were divided into 3 cohorts: the first was treated with levofloxacin, the second with cotrimoxazol and the third was a subgroup without antibiotic prophylaxis (control group). The incidence of infection, rate of mortality, and reduction of hospitalization rate for treatment with parenteral antibiotics were tested. RESULTS: There was a reduction in documented infections (clinically or microbiologically) when comparing the levofloxacin cohort with the control cohort (p < 0.0001) and the levofloxacin cohort with the cotrimoxazol group (p < 0.01). The reduction in the hospitalization rate for treatment with parenteral antibiotics reached statistical significance when comparing the levofloxacin group with the control cohort (p < 0.001) and levofloxacin group with the cotrimoxazol group (p < 0.05). Although the rate of global mortality was lower in the levofloxacin group than in the other two groups, no statistical significance was observed. CONCLUSIONS: Our results show that levofloxacin effectively reduces the incidence of infection, the rate of hospitalization and the requirement for parenteral antibiotics. Although we found a reduction in the overall mortality and in the infection-related mortality, the corresponding data did not reach statistical significance. PMID- 12646108 TI - [Effect of pneumoperitoneum on venous hemodynamics during laparoscopic cholecystectomy. Influence of patients' age and time of surgery]. AB - BACKGROUND AND OBJECTIVE: Abdominal hyperpressure developed during laparoscopic cholecystectomy by the effect of pneumoperitoneum represents an obstacle to the venous return that may facilitate thromboembolic complications. The aim of this study was to establish the effect of pneumoperitoneum in venous hemodynamics during laparoscopy. PATIENTS AND METHOD: Prospective study of 31 consecutive patients who underwent laparoscopic cholecystectomy. Venous occlusion plethysmography was performed preoperatively, after anaesthetic induction, after insufflation, before pneumoperitoneum release and at the end of surgery. Changes of plethysmography were compared with preoperative values and according to age, obesity, presence of varicose veins and pneumoperitoneum time. Bilateral lower limb venous Duplex scanning was performed at days 1, 7 and 30 to detect deep venous thrombosis (DVT). RESULTS: Average age was 56 years, 66.6% females, 40% obese, 16% with varicose veins and the pneumoperitoneum time was < 45 min in 22.5% patients. Capacitance decreased progressively during surgery and was significantly reduced with pneumoperitoneum. The maximum venous outflow in the first second was reduced significantly at the end of pneumoperitoneum. These reductions were more evident in older patients. CONCLUSIONS: Pneumoperitoneum produces plethysmographic changes on venous hemodynamics with a diminished venous return in lower limbs. Older patients have higher risk of thromboembolic complications, while obesity can also increase this risk. However, no DVT was demonstrated in this study. PMID- 12646109 TI - [Taking decisions at the end of life. A survey of the attitude of critically ill patients against death]. AB - BACKGROUND AND OBJECTIVE: An attitude of refusal regarding death still persists in our society and the majority of patients do not usually take decisions related to the end of life. SUBJECTS AND METHOD: From June 1st to December 31st, 2000, 88 first-degree relatives were interviewed concerning patient's preparation before his/her death. Patients considered to be candidates to take decisions about end of life were those with poor quality of life and previously admitted to the intensive care unit (ICU). Relatives were interviewed regarding organ donation, vital willingness, legal representative, civil willingness and patient's knowledge of disease and wish to limit therapeutic efforts in case of extreme severity. Previous quality of life was measured according to the Karnofsky scale. RESULTS: 20% patients had donated organs, none had vital willingness and 44% had civil willingness. When patients were divided into groups of those with and without legal willingness, vital representative, and willgness to donate organs and to limit therapeutic effort, there were no statistically significant differences except for patients with vital representatives who were significantly younger. CONCLUSIONS: Society needs to advance progressively towards the acceptance and preparation of death, so that decisions taken at the end of life, particularly in the ICUs, may represent, as accurately as possible, a patient's scale of values. PMID- 12646110 TI - [Low prevalence of the Leu7/Pro polymorphism in the signal peptide of preproneuropeptide Y in the Spanish Mediterranean population]. AB - BACKGROUND AND OBJECTIVE: A common Leu7Pro polymorphism in the signal peptide of neuropeptide Y has been associated with increased plasma cholesterol and accelerated atherosclerosis in the Finnish population. SUBJECTS AND METHOD: A cross-sectional design was carried out in 1,000 individuals from the Mediterranean Spanish population to estimate its prevalence. Two independently collected samples consisting of 486 subjects (sample 1) and 514 subjects (sample 2) were studied. The Leu7Pro polymorphism was determined. RESULTS: We found only one individual carrying the Leu7Pro variant (0.1%). The allele frequency of the Pro7 allele was extremely low: 0.005 (95% CI,0.0001-0.0015). CONCLUSIONS: This low prevalence suggests that the Pro7 allele is not one of the genetic determinant factors affecting obesity, plasma lipids or alcoholism in this population. PMID- 12646111 TI - [Heading for systematic flu vaccination?]. PMID- 12646112 TI - [Laboratory's function in clinic practice of breast cancer]. PMID- 12646113 TI - [MALT gastric lymphoma]. PMID- 12646114 TI - [Bone lesions in a 59-year-old male]. PMID- 12646115 TI - [Meningoencephalitis caused by HHV-6A in a previously healthy immunocompetent adult]. PMID- 12646116 TI - [Usefulness of autologous transfusion and erythropoietin in surgical and critically ill patients]. PMID- 12646117 TI - [Percutaneous fentanyl in fibromyalgia]. PMID- 12646118 TI - [Anterior uveitis in a patient treated with pegylated interferon and ribavirin]. PMID- 12646119 TI - Cough, primary care and other matters. PMID- 12646120 TI - Clinical efficacy and safety of a depigmented and glutaraldehyde polymerized therapeutic vaccine of Parietaria judaica. AB - BACKGROUND: The inhalation of Parietaria judaica pollen is a common cause of allergic respiratory diseases in the Mediterranean area. The objective of this study was to investigate the safety and clinical efficacy of a chemically modified (depigmented and glutaraldehyde-polymerized) vaccine of Parietaria judaica. METHODS AND RESULTS: Thirty patients with a well-documented clinical history of seasonal rhinitis and clinical sensitivity to Parietaria judaica pollen were included in a randomized trial during 12 months. The study was conducted following good clinical practices and appropriate consent forms were signed. Patients were divided into 2 groups of 15 individuals; group A received the modified extract and group C did not receive specific immunotherapy. Any adverse event was recorded to assess safety. Symptom scores, symptomatic medication use and the results of specific nasal challenges (before and after 12 months of treatment) were recorded to evaluate clinical efficacy. The treatment schedule consisted of an incremental phase of 5 injections and a maintenance dosage of 0.5 ml per month. Each patient received 14 injections during this period. All the patients completed the trial and no adverse reactions related to immunotherapy were recorded. A significant difference (p < 0.001) in symptom scores and overall use of symptomatic medication was observed between the two groups, being both scores lower in group A. No significant differences in nasal sensitivity existed before treatment among the 2 groups. However, after 12 months, a significant difference (p < 0.05) was observed only in group A patients, who showed a significant improvement in specific nasal challenges. CONCLUSIONS: Immunotherapy with depigmented and glutaraldehyde-polymerized extract of Parietaria judaica pollen is safe and effective to treat patients with allergic rhinitis and clinical sensitivity to this pollen. PMID- 12646121 TI - [Hypersensitiviy to Trichophyton rubrum antigens in atopic and non-atopic podiatrists]. AB - BACKGROUND: Podiatrists as well as manicurists are exposed to inhale nail dust contaminated with mycotic particles when filing and burring the onychomycotic nails of their patients. As some of them with atopic background suffered worsening of their symptoms we decided to study the immune response to the fungus Trichophyton rubrum (Tr) that was isolated from the nail dust obtained from the podiatrists office. METHODOLOGY AND RESULTS: A detailed clinical record, intracutaneous skin tests with a extract of Tr and the serum levels of total and specific IgE were performed in rhinitis-asthma as well as control patients. As the asthmatic group refused to perform the challenge bronchial test with Tr we developed a guinea pig experimental model with daily aerosolization of Tr during 12 weeks studying the levels of specific IgE and IgG as well as the lung's histopathology. Atopic patients showed positive immediate skin tests with Tr and both groups revealed delayed hypersensitivity to the antigen.RAST-IgE-anti-Tr and RAST-inhibition confirmed the specificity of the antibodies. Guinea pigs also synthetized IgG and IgE anti-Tr and suffered different degrees of lung lesions similar to those of hypersensitivity pneumonitis. CONCLUSION: Atopic podiatrists are exposed to fungal allergens that may participate or aggravate their previous respiratory conditions. PMID- 12646122 TI - Single-course specific immunotherapy with mixed pollen allergoids: results of a multi-centre study. AB - BACKGROUND: A short-term immunotherapy vaccine for the treatment of pollen allergy has been developed utilising L-tyrosine adsorbed allergoids. The reduced number of injections could provide advantages over long-term therapy schedules. This would improve compliance and support application of specific immunotherapy (SIT) to a greater extent. We report a multicenter study to evaluate the efficacy and safety of this treatment in a clinical practice setting. METHODS: Patients (n = 1808) with a diagnosis of sensitivities to various pollens and symptoms of allergic asthma and/or allergic rhinitis and/or allergic conjunctivitis were selected. The vaccine formulation was made up according to individual sensitivities and contained L-tyrosine adsorbed allergoids. The patients were treated with a 3-injection initial course followed by a 3-injection maintenance course. Efficacy was measured by consumption of symptomatic anti-allergic medication compared with that in the previous season and by physician assessment using a 5-point scale. All adverse events were recorded. RESULTS: Efficacy was demonstrated by a considerable decrease in regular and frequent use of medication compared with that in the previous season (p < 0.001). In addition, in 80 % of the patients, the physician's assessment was either "good" or "very good". These outcomes were unaffected by the closeness of the treatment course to the onset of the pollen season. Tolerability was good and most local and systemic reactions were mild. CONCLUSIONS: The treatment of pollen-allergic patients with a short term SIT using a 6-injection pollen allergoid/L-tyrosine vaccine in a clinical practice setting provided a high level of efficacy with a low incidence of mainly mild adverse events. PMID- 12646123 TI - Serum IgE level in malnutrition. AB - Infections and malnutrition remain the main causes of infant mortality in developing countries. In protein-calorie malnutrition, immunologic responses are affected, which often facilitates infections. However, the presence of asthma and allergic rhinitis are not commonly recognized in malnourished individuals. The aim of this study was to evaluate serum IgE values in children with primary moderate protein-calorie malnutrition. METHODS: The level of IgE in peripheral blood of 18 children between 2 and 4 old with moderate protein-calorie malnutrition and without associated parasitic infestation was compared with that of 15 well nourished children of similar age. IgE serum levels were measured by an immunoenzymatic method. RESULTS: The median level of serum IgE in malnourished children was 69.30 ng/ml while the control group showed a mean level of 95.97 ng/ml. This difference was significant (p < 0.01). CONCLUSION: Malnourished children show decreased serum IgE levels. This might be one of the adaptive mechanisms of malnutrition employed in an attempt to use energy and protein reserves for growth and other functions. Our results are coherent with the decrease in IgE mediated reactions in malnourished patients. PMID- 12646125 TI - A case of anaphylaxis due to rose pollen ingestion. AB - BACKGROUND: Recent publications have demonstrated that in Guneykent (Turkey), a rose-cultivating area, some workers experience various allergic reactions due to contact with rose or its pollen. CASE REPORT: A 47-year-old man, originally from Guneykent although he no longer lived there, suffered perennial rhinitis, asthma and rhinoconjunctivitis in spring related to sensitization to dust mites and various pollens, respectively. On several occasions, he had presented oral and cutaneous symptoms and angioedema due to contact with rose or intake of honey or other products. Finally, he had an anaphylactic reaction on drinking dew collected in the tulips of a rose that possibly contained rose pollen. METHODS: Specific IgE was evaluated with Rosa rugosa extract and was 30.3 IU (class IV). Sensitization to other pollens and to mites was also confirmed. Oral provocation test was not deemed ethical. CONCLUSION: This may be the first reported case of anaphylactic reaction due to rose pollen ingestion. PMID- 12646124 TI - Knowledge of pediatricians in Rio de Janeiro, Brazil, about inhalation therapy in asthmatic children. AB - BACKGROUND: Despite the numerous guidelines on the diagnosis and treatment of asthma, there are data that indicate that general pediatrician's knowledge of the disease and its preferred treatment is limited, which may influence the quality of care given to asthmatic children. The purpose of this study was to describe pediatrician's knowledge of spacers and of concepts of chilhood asthma, as well as their prescribing habits concerning inhalation therapy, in the public health system of the city of Rio de Janeiro. METHODS AND RESULTS: A descriptive cross sectional study was performed in a sample of 72 pediatricians from the public health system of Rio de Janeiro. A questionnaire was used to assess prescriptions for spacers, the reasons whay spacers were not used, the models employed, classification of asthma according to clinical severity and symptom frequency, recommendation for the correct spacer volume according to age group, and the concept of asthma as an inflammatory disease. Seventy percent (51/72) of the physicians did not routinely prescribe the spacer. The reasons given were as follows: a) lack of spacer availability in the public health system in 55 % (28/51); b) high cost in 57 % (29/51); c) the complexity of their use in 35 % (18/51); d) unwillingness to use aerosol type medication in 15 % (8/51), and e)lack of knowledge of their function and utilization in 59 % (30/51). Of the 30 % (21/72) who reported they regularly and routinely prescribed the spacer in daily practice, 48 % (10/21) stated that this routine prescription, even when indicated, was below 25 % of what was expected and makeshift models were preferred by 24 % (5/21) of the pediatricians. Six percent of the pediatricians chose the appropriate spacer volume according to age group, 62.5 % (45/72) reported that they classified asthma according to severity, 16 % (7/45) gave the correct answers when classifying asthma according to national consensus, and 22 % (16/72) considered asthma to be an inflammatory disease. CONCLUSIONS: The results of this study suggest that pediatrician's knowledge of inhalation therapy with dosed aerosol spacers and of asthma-related concepts in the public health system in Rio de Janeiro is limited. This may restrict the quality of care given to the asthmatic children in the city and suggests the need for training programs for the management of asthmatic children. PMID- 12646126 TI - Successful desensitization to penicillin after diagnostic reassessment. AB - Clinical situations for which penicillin is indicated as the sole effective treatment are not infrequent, in these circumstances penicillin allergy complicates their medical management. No proven alternatives to penicillin are available for treating neurosyphilis, congenital syphilis, or syphilis in pregnant women, it is not possible choosing an alternative non betalactam antibiotic. The management of the patient who has a history of penicillin allergy include skin testing to determine if penicillin-specific IgE antibodies exist, and desensitization if penicillin is the choice treatment. We report a case of successful desensitization to penicillin in a 30 year-old pregnant woman with latent syphilis and penicillin allergy, where the first penicillin study was not positive, but the repetition of diagnosis test (reassessment) with the same betalactam reagents than in the first study two weeks later the initial evaluation was positive, detecting allergic sensitization not detected at the previous examination. PMID- 12646127 TI - A WASp homolog powers actin polymerization-dependent motility of endosomes in vivo. AB - BACKGROUND: WASp/SCAR proteins activate the Arp2/3 complex to nucleate actin filament assembly and are thought to have important roles in endocytosis. WASp is required for efficient endocytosis of antigen receptors, N-WASp promotes actin polymerization-dependent movement of endomembrane vesicles, and Las17 (a yeast WASp homolog) is required for endocytic internalization. However, it is unknown whether movement of endosomes or other organelles requires activation of the Arp2/3 complex by members of the WASp/SCAR family. RESULTS: Fluorescence video microscopy of yeast cells expressing a GFP-tagged G protein-coupled receptor (Ste2-GFP) as an endocytic marker revealed that endosomes and the lysosome-like vacuole are highly motile. Endosome/vacuole motility required actin polymerization, as indicated by sensitivity to latrunculin A, whereas microtubules were uninvolved. Endosome/vacuole motility did not require actin cables or myosin V (a MYO2 gene product), which moves secretory vesicles and the Golgi apparatus and mediates vacuole segregation. However, endosome motility required Las17, a WASp homolog. In contrast to other processes involving Las17, endosome/vacuole motility required the WCA domain of Las17, which is necessary and sufficient to activate the Arp2/3 complex. CONCLUSIONS: Endosome/vacuole motility in vivo requires actin polymerization stimulated by the WASp homolog Las17. WASp/SCAR family members in mammalian cells may have similar functions. Defects in endosome/lysosome motility may contribute to deficits in lymphocyte or macrophage function observed in human patients lacking WASp or developmental defects in N-WASp-deficient mice. PMID- 12646128 TI - African Y chromosome and mtDNA divergence provides insight into the history of click languages. AB - BACKGROUND: About 30 languages of southern Africa, spoken by Khwe and San, are characterized by a repertoire of click consonants and phonetic accompaniments. The Jumid R:'hoansi (!Kung) San carry multiple deeply coalescing gene lineages. The deep genetic diversity of the San parallels the diversity among the languages they speak. Intriguingly, the language of the Hadzabe of eastern Africa, although not closely related to any other language, shares click consonants and accompaniments with languages of Khwe and San. RESULTS: We present original Y chromosome and mtDNA variation of Hadzabe and other ethnic groups of Tanzania and Y chromosome variation of San and peoples of the central African forests: Biaka, Mbuti, and Lisongo. In the context of comparable published data for other African populations, analyses of each of these independently inherited DNA segments indicate that click-speaking Hadzabe and Jumid R:'hoansi are separated by genetic distance as great or greater than that between any other pair of African populations. Phylogenetic tree topology indicates a basal separation of the ancient ancestors of these click-speaking peoples. That genetic divergence does not appear to be the result of recent gene flow from neighboring groups. CONCLUSIONS: The deep genetic divergence among click-speaking peoples of Africa and mounting linguistic evidence suggest that click consonants date to early in the history of modern humans. At least two explanations remain viable. Clicks may have persisted for tens of thousands of years, independently in multiple populations, as a neutral trait. Alternatively, clicks may have been retained, because they confer an advantage during hunting in certain environments. PMID- 12646129 TI - A function for Egf receptor signaling in expanding the developing brain in Drosophila. AB - BACKGROUND: In invertebrates and vertebrates, neural midline cells secrete signals that pattern the central nervous system (CNS). However, an important part of the developing insect brain, involved in functions such as olfaction and feeding behavior, is positioned lateral to the foregut and lacks neural cells at the midline. Could the foregut substitute for neural midline cells and secrete signals that pattern this part of the brain? RESULTS: In Drosophila embryos, the neural midline marker Single-minded is expressed in foregut cells adjacent to the brain, as are members of the Egf receptor signaling pathway. Removing the function of these molecules results in aberrant proliferation and reduced size in the brain lateral to the foregut. CONCLUSIONS: Cells of the brain lateral to the foregut receive an Egf signal from the midline and proliferate in response. A likely source of this signal is the foregut. These findings raise the possibility that the brain lateral to the foregut is an evolutionarily recent addition to the arthropod brain, and that the anterior boundary of the brain neural midline is a conserved feature in bilaterally symmetric animals. PMID- 12646130 TI - Viewing geometry determines how vision and haptics combine in size perception. AB - Vision and haptics have different limitations and advantages because they obtain information by different methods. If the brain combined information from the two senses optimally, it would rely more on the one providing more precise information for the current task. In this study, human observers judged the distance between two parallel surfaces in two within-modality experiments (vision alone and haptics-alone) and in an intermodality experiment (vision and haptics together). In the within-modality experiments, the precision of visual estimates varied with surface orientation, as expected from geometric considerations; the precision of haptic estimates did not. An ideal observer that combines visual and haptic information weights them differently as a function of orientation. In the intermodality experiment, humans adjusted visual and haptic weights in a fashion quite similar to that of the ideal observer. As a result, combined size estimates are finer than is possible with either vision or haptics alone; indeed, they approach statistical optimality. PMID- 12646131 TI - Maternal transfer of strain-specific immunity in an invertebrate. AB - The most celebrated component of the vertebrate immune system is the acquired response in which memory cells established during primary infection enhance the proliferation of antibodies during secondary infection. Additionally, the strength of vertebrate acquired immune responses varies dramatically depending on the infecting pathogen species or on the pathogen genotype within species. Because invertebrates lack the T-cell receptors and Major Histocompatibility Complex (MHC) molecules that mediate vertebrate adaptive immune responses, they are thought to lack adaptive immunity and be relatively unspecific in their interactions with pathogens. With only innate immunity, invertebrate hosts are believed to be nai;ve at each new encounter with pathogens. Nevertheless, some forms of facultative immunity appear to be important in insects; some individuals have enhanced immunity due to population density, and some social insects benefit when their nest-mates have been exposed to a pathogen or pathogen mimic (; see for a predation example.) Here we provide evidence for acquired strain-specific immunity in the crustacean Daphnia magna infected with the pathogenic bacteria Pasteuria ramosa. Specifically, the fitness of hosts was enhanced when challenged with a bacterial strain their mother had experienced relative to cases when mother and offspring were challenged with different strains. PMID- 12646132 TI - The cost of cortical computation. AB - Electrophysiological recordings show that individual neurons in cortex are strongly activated when engaged in appropriate tasks, but they tell us little about how many neurons might be engaged by a task, which is important to know if we are to understand how cortex encodes information. For human cortex, I estimate the cost of individual spikes, then, from the known energy consumption of cortex, I establish how many neurons can be active concurrently. The cost of a single spike is high, and this severely limits, possibly to fewer than 1%, the number of neurons that can be substantially active concurrently. The high cost of spikes requires the brain not only to use representational codes that rely on very few active neurons, but also to allocate its energy resources flexibly among cortical regions according to task demand. The latter constraint explains the investment in local control of hemodynamics, exploited by functional magnetic resonance imaging, and the need for mechanisms of selective attention. PMID- 12646133 TI - A model system for increased meiotic nondisjunction in older oocytes. AB - For at least 5% of all clinically recognized human pregnancies, meiotic segregation errors give rise to zygotes with the wrong number of chromosomes. Although most aneuploid fetuses perish in utero, trisomy in liveborns is the leading cause of mental retardation. A large percentage of human trisomies originate from segregation errors during female meiosis I; such errors increase in frequency with maternal age. Despite the clinical importance of age-dependent nondisjunction in humans, the underlying mechanisms remain largely unexplained. Efforts to recapitulate age-dependent nondisjunction in a mammalian experimental system have so far been unsuccessful. Here we provide evidence that Drosophila is an excellent model organism for investigating how oocyte aging contributes to meiotic nondisjunction. As in human oocytes, nonexchange homologs and bivalents with a single distal crossover in Drosophila oocytes are most susceptible to spontaneous nondisjunction during meiosis I. We show that in a sensitized genetic background in which sister chromatid cohesion is compromised, nonrecombinant X chromosomes become vulnerable to meiotic nondisjunction as Drosophila oocytes age. Our data indicate that the backup pathway that normally ensures proper segregation of achiasmate chromosomes deteriorates as Drosophila oocytes age and provide an intriguing paradigm for certain classes of age-dependent meiotic nondisjunction in humans. PMID- 12646134 TI - Phosphatidylinositol-5-phosphate activation and conserved substrate specificity of the myotubularin phosphatidylinositol 3-phosphatases. AB - Phosphoinositides control many different processes required for normal cellular function. Myotubularins are a family of Phosphatidylinositol 3-phosphate (PtdIns3P) phosphatases identified by the positional cloning of the MTM1 gene in patients suffering from X-linked myotubular myopathy and the MTMR2 gene in patients suffering from the demyelinating neuropathy Charcot-Marie-Tooth disease type 4B. MTM1 is a phosphatidylinositol phosphatase with reported specificity toward PtdIns3P, while the related proteins MTMR2 and MTMR3 hydrolyze both PtdIns3P and PtdIns(3,5)P2. We have investigated MTM1 and MTMR6 and find that they use PtdIns(3,5)P2 in addition to PtdIns3P as a substrate in vitro. The product of PtdIns(3,5)P2 hydrolysis, PtdIns5P, causes MTM1 to form a heptameric ring that is 12.5 nm in diameter, and it is a specific allosteric activator of MTM1, MTMR3, and MTMR6. A disease-causing mutation at arginine 69 of MTM1 falling within a putative pleckstrin homology domain reduces the ability of the enzyme to respond to PtdIns5P. We propose that the myotubularin family of enzymes utilize both PtdIns3P and PtdIns(3,5)P2 as substrates, and that PtdIns5P functions in a positive feedback loop controlling their activity. These findings highlight the importance of regulated phosphatase activity for the control of phosphoinositide metabolism. PMID- 12646135 TI - Dynamin 3 is a component of the postsynapse, where it interacts with mGluR5 and Homer. AB - The dynamins comprise a large family of mechanoenzymes known to participate in membrane modeling events. All three conventional dynamin genes (Dyn1, Dyn2, Dyn3) are expressed in mammalian brain and produce more than 27 different dynamin proteins as a result of alternative splicing. Past studies have suggested that Dyn1 participates in specialized neuronal functions such as rapid synaptic vesicle recycling, while Dyn2 may mediate the conventional clathrin-mediated uptake of surface receptors. Currently, the distribution, expression, and function of Dyn3 in neurons, or in any other cell type, are completely undefined. Here, we demonstrate that Dyn1 and Dyn3 localize differentially in the synapse. Dyn1 concentrates within the presynaptic compartment, while Dyn3 localizes to dendritic spine tips. Within the postsynaptic density (PSD), we found Dyn3, but not Dyn1, to be part of a biochemically isolated complex comprised of Homer and metabotropic glutamate receptors. Finally, although dominant-negative Dyn3 did not seem to inhibit receptor endocytosis, overexpression of a specific Dyn3 spliced variant in mature neurons caused a marked remodeling of dendritic spines. These data suggest that Dyn3 is a postsynaptic dynamin and, like its binding partner Homer, plays a significant role in dendritic spine morphogenesis and remodeling. PMID- 12646136 TI - Hyperactivation of the G12-mediated signaling pathway in Caenorhabditis elegans induces a developmental growth arrest via protein kinase C. AB - The G(12) type of heterotrimeric G-proteins play an important role in development and behave as potent oncogenes in cultured cells. However, little is known about the molecular nature of the components that act in the G(12)-signaling pathway in an organism. We characterized a C. elegans Galpha subunit gene, gpa-12, which is a homolog of mammalian G(12)/G(13)alpha, and found that animals defective in gpa 12 are viable. Expression of activated GPA-12 (G(12)QL) results in a developmental growth arrest caused by a feeding behavior defect that is due to a dramatic reduction in pharyngeal pumping. To elucidate the molecular nature of the signaling pathways in which G(12) participates, we screened for suppressors of the G(12)QL phenotype. We isolated 50 suppressors that contain mutations in tpa-1, which encodes two protein kinase C isoforms, TPA-1A and TPA-1B, most similar to PKCtheta/delta. TPA-1 mediates the action of the tumor promoter PMA. Expression of G(12)QL and treatment of wild-type animals with PMA induce an identical growth arrest caused by inhibition of larval feeding, which is dependent on TPA-1A and TPA-1B function. These results suggest that TPA-1 is a downstream target of both G(12) signaling and PMA in modulating feeding and growth in C. elegans. Taken together, our findings provide a potential molecular mechanism for the transforming capability of G(12) proteins. PMID- 12646137 TI - An interference effect of observed biological movement on action. AB - It has been proposed that actions are intrinsically linked to perception and that imagining, observing, preparing, or in any way representing an action excites the motor programs used to execute that same action. There is neurophysiological evidence that certain brain regions involved in executing actions are activated by the mere observation of action (the so-called "mirror system;" ). However, it is unknown whether this mirror system causes interference between observed and simultaneously executed movements. In this study we test the hypothesis that, because of the overlap between action observation and execution, observed actions should interfere with incongruous executed actions. Subjects made arm movements while observing either a robot or another human making the same or qualitatively different arm movements. Variance in the executed movement was measured as an index of interference to the movement. The results demonstrate that observing another human making incongruent movements has a significant interference effect on executed movements. However, we found no evidence that this interference effect occurred when subjects observed a robotic arm making incongruent movements. These results suggest that the simultaneous activation of the overlapping neural networks that process movement observation and execution infers a measurable cost to motor control. PMID- 12646138 TI - Circadian control of eclosion: interaction between a central and peripheral clock in Drosophila melanogaster. AB - Drosophila melanogaster display overt circadian rhythms in rest:activity behavior and eclosion. These rhythms have an endogenous period of approximately 24 hr and can adjust or "entrain" to environmental inputs such as light. Circadian rhythms depend upon a functioning molecular clock that includes the core clock genes period and timeless (reviewed in and ). Although we know that a clock in the lateral neurons (LNs) of the brain controls rest:activity rhythms, the cellular basis of eclosion rhythms is less well understood. We show that the LN clock is insufficient to drive eclosion rhythms. We establish that the prothoracic gland (PG), a tissue required for fly development, contains a functional clock at the time of eclosion. This clock is required for normal eclosion rhythms. However, both the PG clock function and eclosion rhythms require the presence of LNs. In addition, we demonstrate that pigment-dispersing factor (PDF), a neuropeptide secreted from LNs, is necessary for the PG clock and eclosion rhythms. Unlike other clocks in the fly periphery, the PG is similar to mammalian peripheral oscillators because it depends upon input, including PDF, from central pacemaker cells. This is the first report of a peripheral clock necessary for a circadian event. PMID- 12646139 TI - Death of Dolly marks cloning milestone. AB - Dolly the cloned sheep marked both an icon of scientific possibility and a potential ethical nightmare. Researchers in related fields of stem cell research need to learn the lessons and the right vocabulary if they are to make progress in their fields. PMID- 12646140 TI - Click to connect. AB - Humans have been enormously successful in exploiting a range of sounds used in language, but none more so than the click consonants used by a few populations in eastern and southern Africa. New research suggests these click languages represent an ancient human language. PMID- 12646141 TI - Martin Nowak. PMID- 12646143 TI - APC. PMID- 12646144 TI - Receptive fields. PMID- 12646145 TI - Vomeronasal organ: pheromone recognition with a twist. AB - Pheromones are detected by the vomeronasal organ using members of two receptor superfamilies: the V1Rs and V2Rs. New studies show that MHC class I molecules are co-expressed in particular combinations with specific V2Rs in the vomeronasal organ. The role of these MHC molecules is unknown, but they may be of considerable biological significance. PMID- 12646146 TI - Gastrulation: PARtaking of the bottle. AB - Gastrulation in C. elegans embryos involves ingression of individual cells, but is driven by apical constriction of the kind that promotes migration of epithelial cell sheets. Recent work shows that PAR proteins, known for their role in polarization and unequal cell division, are also associated with the polarization that establishes this apical constriction. PMID- 12646148 TI - Gene duplications: the gradual evolution of functional divergence. AB - Budding yeast provides a useful resource for studies of gene function. A new analysis of the fitness effects of deletion mutations in budding yeast reveals that genes that have duplicates create lower fitness losses when inactivated than do genes that are singletons. PMID- 12646147 TI - Sensory biology: how the nose knows. AB - Sensory information is encoded as patterns of synaptic activity. Recent evidence suggests that differential synaptic release and use of postsynaptic glutamate receptors is critical for encoding information from polymodal neurons. PMID- 12646149 TI - Fragile sites: breaking up over a slowdown. AB - Common fragile sites are specific regions in the human genome that are particularly prone to genomic instability under conditions of replicative stress. Recent data suggest that these sites depend on the checkpoint kinase ATR to maintain their stability. PMID- 12646150 TI - Shape perception: complex contour representation in visual area V4. AB - A recent study has put forward a physiologically plausible population model that implements a parts-based shape-coding scheme for macaque visual area V4. PMID- 12646151 TI - Lamellipodia protrusion: moving interactions of vinculin and Arp2/3. AB - A direct, transient interaction between vinculin and Arp2/3 is required to promote receptor-stimulated lamellipodial extension and cell spreading. Vinculin selectively recruits Arp2/3 to the leading edge of the lamellipodium, where it may couple the actin polymerization machinery to adhesion complexes to promote membrane protrusion over ruffling. PMID- 12646152 TI - Receptor sensitivity: when T cells lose their sense of self. AB - The T-cell antigen receptor binds to self-MHC molecules with low affinity. Recent reports disagree as to whether this interaction sensitizes or desensitizes the receptor. Here we discuss how these findings might be reconciled. PMID- 12646153 TI - Molecular links between X-inactivation and autosomal imprinting: X-inactivation as a driving force for the evolution of imprinting? AB - In classical Mendelian inheritance, each parent donates a set of chromosomes to its offspring so that maternally and paternally encoded information is expressed equally. The phenomena of X-chromosome inactivation (XCI) and autosomal imprinting in mammals violate this dogma of genetic equality. In XCI, one of the two female X chromosomes is silenced to equalize X-linked gene dosage between XX and XY individuals. In genomic imprinting, parental marks determine which of the embryo's two autosomal alleles will be expressed. Although XCI and imprinting appear distinct, molecular evidence now shows that they share a surprising number of features. Among them are cis-acting control centers, long-distance regulation and differential DNA methylation. Perhaps one of the most intriguing similarities between XCI and imprinting has been their association with noncoding and antisense RNAs. Very recent data also suggest the common involvement of histone modifications and chromatin-associated factors such as CTCF. Collectively, the evidence suggests that XCI and genomic imprinting may have a common origin. Here, I hypothesize that the need for X-linked dosage compensation was a major driving force in the evolution of genomic imprinting in mammals. I propose that imprinting was first fixed on the X chromosome for XCI and subsequently acquired by autosomes. PMID- 12646154 TI - Learning about life and death in early childhood. AB - have argued that young children initially understand biological phenomena in terms of vitalism, a mode of construal in which "life" or "life-force" is the central causal-explanatory concept. This study investigated the development of vitalistic reasoning in young children's concepts of life, the human body and death. Sixty preschool children between the ages of 3 years, 7 months and 5 years, 11 months participated. All children were initially given structured interviews to assess their knowledge of (1) human body function and (2) death. From this sample 40 children in the Training group were taught about the human body and how it functions to maintain life. The Control group (n=20) received no training. All 60 children were subsequently reassessed on their knowledge of human body function and death. Results from the initial interviews indicated that young children who spontaneously appealed to vitalistic concepts in reasoning about human body functioning were also more sophisticated in their understanding of death. Results from the posttraining interviews showed that children readily learned to adopt a vitalistic approach to human body functioning, and that this learning coincided with significant development in their understanding of human body function, and of death. The overall pattern of results supports the claim that the acquisition of a vitalistic causal-explanatory framework serves to structure children's concepts and facilitates learning in the domain of biology. PMID- 12646155 TI - Motion and edge sensitivity in perception of object unity. AB - Although much evidence indicates that young infants perceive unitary objects by analyzing patterns of motion, infants' abilities to perceive object unity by analyzing Gestalt properties and by integrating distinct views of an object over time are in dispute. To address these controversies, four experiments investigated adults' and infants' perception of the unity of a center-occluded, moving rod with misaligned visible edges. Both alignment information and depth information affected adults' and infants' perception of object unity in similar ways, and infants perceived object unity by integrating information about object features over time. However, infants perceived a moving, misaligned, three dimensional object as indeterminate in its connectedness, whereas adults perceived it as connected behind the occluder. These findings indicate that the effectiveness of common motion in specifying unified surfaces across an occluder is reduced by misalignment of edges. Alignment information enhances perception of object unity either by serving directly as information for unity or by optimizing the detectability of motion-carried information for unity. In addition, young infants are able to retain information about edge orientation over short intervals in determining connectedness via a process of spatiotemporal integration. PMID- 12646156 TI - Lexical viability constraints on speech segmentation by infants. AB - The Possible Word Constraint limits the number of lexical candidates considered in speech recognition by stipulating that input should be parsed into a string of lexically viable chunks. For instance, an isolated single consonant is not a feasible word candidate. Any segmentation containing such a chunk is disfavored. Five experiments using the head-turn preference procedure investigated whether, like adults, 12-month-olds observe this constraint in word recognition. In Experiments 1 and 2, infants were familiarized with target words (e.g., rush), then tested on lists of nonsense items containing these words in "possible" (e.g., "niprush" [nip+rush]) or "impossible" positions (e.g., "prush" [p+rush]). The infants listened significantly longer to targets in "possible" versus "impossible" contexts when targets occurred at the end of nonsense items (rush in "prush"), but not when they occurred at the beginning (tan in "tance"). In Experiments 3 and 4, 12-month-olds were similarly familiarized with target words, but test items were real words in sentential contexts (win in "wind" versus "window"). The infants listened significantly longer to words in the "possible" condition regardless of target location. Experiment 5 with targets at the beginning of isolated real words (e.g., win in "wind") replicated Experiment 2 in showing no evidence of viability effects in beginning position. Taken together, the findings suggest that, in situations in which 12-month-olds are required to rely on their word segmentation abilities, they give evidence of observing lexical viability constraints in the way that they parse fluent speech. PMID- 12646157 TI - Protein prenylation: a pivotal posttranslational process. PMID- 12646158 TI - Incorporation of DUF/FACT into chromatin enhances the accessibility of nucleosomal DNA. AB - DNA unwinding factor (DUF) was discovered as an essential DNA replication factor in Xenopus egg extracts. DUF consists of an HMG protein and a homolog of Cdc68p/Spt16p, and has the capability of unwinding dsDNA. Here we have examined the interaction of DUF with chromatin. DUF was incorporated into chromatin assembled from sperm heads and from plasmid DNA in egg extracts. It was revealed that the chromatin assembled in egg extracts immunodepleted of DUF is less sensitive to micrococcal nuclease (NNase) digestion than that assembled in control extracts, indicating that chromatin containing DUF has more decompact structure than that without DUF. Also we found that DUF has a high affinity for core histones in vitro. We suggest that the function of DUF may be to make the chromatin structure accessible to replication factors. PMID- 12646159 TI - Elevated plasma deoxyuridine in patients with thymidine phosphorylase deficiency. AB - Mutations in the nuclear gene encoding thymidine phosphorylase (TP) cause mitochondrial neurogastrointestinal encephalomyopathy (MNGIE), an autosomal recessive disease with mitochondrial dysfunction and mitochondrial DNA abnormalities. We have demonstrated alterations of thymidine (dThd) metabolism in MNGIE patients. Here, we report the accumulation of another substrate of TP, deoxyuridine (dUrd), whose circulating levels ranged from 5.5 to 24.4 microM (average 14.2) in MNGIE and were undetectable (<0.05 microM) in both TP mutation carriers and controls. The dramatic accumulation of dUrd may contribute to nucleotide pool imbalances and, together with the increased levels of dThd, is likely to contribute to the pathogenesis of MNGIE. PMID- 12646160 TI - Active transducin alpha subunit carries PDE6 to detergent-resistant membranes in rod photoreceptor outer segments. AB - cGMP-Phosphodiesterase 6 (PDE6) is the central effector enzyme in the phototransduction system of vertebrate photoreceptors. We have recently found that PDE6 accumulates in a detergent-resistant membrane (DRM) fraction in response to excitation of bovine rod phototransduction system. Here, we studied the molecular mechanism of the PDE6 translocation to DRM. Pertussis toxin inhibited the translocation of PDE6. Upon addition of AlF(4)(-) to dark-adapted ROS, PDE6 translocated to DRM along with a minor fraction of the alpha subunit of transducin (T alpha). The addition of an excess of the inhibitory subunit of PDE6 blocked its accumulation in the DRM, but did not block the translocation of the minor fraction of T alpha. These data suggested that the formation of a complex between activated T alpha and PDE6 imparted upon T alpha a high affinity for the DRM. The translocation of PDE6 to the DRM may be involved in the spatiotemporal regulation of its activity on disk membranes. PMID- 12646161 TI - Cloning, expression, and characterization of chicken transforming growth factor beta 4. AB - Transforming growth factor beta 4 (TGF-beta 4) is unique to avian species, though its roles in vivo have not yet been well established. In this paper we describe the expression and partial characterization of recombinant chicken TGF-beta 4. By using a GC-rich PCR system in a modified 5'RACE methodology we generated the 5' end of cDNA sequence encoding the TGF-beta 4 precursor, which was in-frame cloned into pcDNA3.1/V5-His-TOPO and transfected into the Chinese hamster ovary cell line (CHO-K1). A cell line stably expressing TGF-beta 4 precursor protein was established from CHO-K1 cells. Acid-activated mature TGF-beta 4 inhibited the growth of mink lung epithelial (Mv1Lu) cell line. TGF-beta 4 also stimulated the expression of type I procollagen and enhanced heat shock protein 47 (Hsp47) expression in chicken tendon fibroblasts. Hsp47 expression by TGF beta 4 is likely regulated through activation of heat shock transcription factor 1 (HSF1). Because the presence of TGF-beta 1 has not been documented in avian cells and our data show that TGF-beta 4 elicits biological activities in chicken tendon cells, which closely parallel that of TGF-beta 1, we propose that TGF-beta 4 plays roles in avian species similar to what TGF-beta 1 plays in mammalian species. PMID- 12646162 TI - Cloning of a novel one-repeat calcium channel-like gene. AB - We describe the cloning of a cDNA from a human testis library that encodes a novel protein with similarity to one repeat of voltage-gated Ca(2+) channels (Ca(v)). Northern and dot blot analyses indicate that the novel Ca(v)-like gene is expressed predominantly in testis and at lower levels in many other tissues. Heterologous expression of the Ca(v)-like protein did not lead to the induction of any detectable ionic current and failed to modify intracellular Ca(2+) concentrations. Similar one-repeat Ca(v)-like proteins have been cloned from Bacillus, Mus, and Homo, and appear to encode ion channels involved in renal function, axis determination, and sperm motility. PMID- 12646164 TI - Function of the loop residue Thr792 in human DNA topoisomerase II alpha. AB - We studied the mutation effect of one of the putative loop residues Thr792 in human DNA topoisomerase II alpha (TOP2 alpha). Thr792 mutants were expressed from high or low copy plasmids in a temperature sensitive yeast strain deficient in TOP2 (top2-1). When expressed from a high copy plasmid, mutants with small side chains complemented the yeast defect; however, from a low copy plasmid, only wild type, Ser, and Cys substitution mutants complemented the yeast defect. Interestingly, at the permissive temperature other mutants (e.g., Val, Gly, and Glu substitutions) showed the dominant negative effect to the top2-1 allele, which was not observed by the control alpha 4-helix mutants. T792E mutant was 10 fold less active than wild-type and the T792P had no decatenation activity in vitro. These results suggest that Thr792 in human TOP2 alpha is involved in enzyme catalysis. PMID- 12646163 TI - Caspase-dependent cleavage of tensin induces disruption of actin cytoskeleton during apoptosis. AB - Members of both calpain and caspase protease families can degrade several components of focal adhesions, leading to disassembly of these complexes. In this report, we investigated the disappearance of tensin from cell adhesion sites of chicken embryonic fibroblast cells (CEFs) exposed to etoposide and demonstrated that loss of tensin from cell adhesions during etoposide-induced apoptosis may be due to degradation of tensin by caspase-3. Tensin cleavage by caspase-3 at the sequence DYPD(1226)G separates the amino-terminal region containing the actin binding domain and the carboxyl-terminal region containing the SH2 domain. The resultant carboxyl-terminal fragment of tensin is unable to bind phosphoinositide 3-kinase (PI3-kinase) transducing cell survival signaling. We also demonstrated that overexpression of the amino-terminal tensin fragment induced disruption of actin cytoskeleton in chicken embryonic fibroblasts. Therefore, caspase-mediated cleavage of tensin contributes to the disruption of actin organization and interrupts ECM-mediated survival signals through phosphatidylinositol 3-kinase. PMID- 12646166 TI - Branched-chain amino acids promote albumin synthesis in rat primary hepatocytes through the mTOR signal transduction system. AB - The administration of branched-chain amino acids (BCAAs) to cirrhosis patients increases serum albumin levels and improves the blood Fischer's ratio. Although it has been reported that albumin synthesis in rat primary hepatocytes is diminished under lower Fisher's ratio conditions compared to normal Fischer's ratio conditions, the mode of action at the molecular level for these effects is still uncertain. It has been reported recently that the triggering signal for protein synthesis is transmitted through mTOR (mammalian target of rapamycin). We have had an interest in the mTOR signal transduction system. In the present study, we analyzed the mode of action of BCAA-induced albumin synthesis using rat primary hepatocytes. The BCAA mixture dose-dependently promoted the production of albumin, with leucine being the major effector half of which was inhibited by the mTOR inhibitor rapamycin. We also showed that only leucine induces P70 S6 kinase activation and 4E-BP1 phosphorylation which are mTOR's downstream translational effectors. These activations were completely inhibited by rapamycin. Our results suggest that BCAAs, especially leucine, promote the production of albumin in rat primary hepatocytes through an mTOR signal transduction system. PMID- 12646165 TI - Calpain-mediated AQP2 proteolysis in inner medullary collecting duct. AB - Vitamin D-elicited hypercalcemia/hypercalciuria is associated with polyuria in humans and in animal models. In rats, dihydrotachysterol (DHT) induces AQP2 water channel downregulation despite unaltered AQP2 mRNA expression and thus we investigated the mechanism of AQP2 degradation. Incubation of AQP2-containing inner medullary collecting duct (IMCD) endosomes with Ca(2+) or calpain elicited AQP2 proteolysis, an effect abolished by leupeptin. This endogenous, Ca(2+) sensitive protease activity exhibited a different proteolytic digest pattern from trypsin, which also degraded AQP2 in vitro. IMCDs contain abundant micro-calpain protein and functional calpain proteolytic activity as demonstrated by immunohistochemistry, immunoblotting, and gel zymography. Furthermore, by small particle flow cytometry we demonstrated that micro-calpain colocalizes with apical IMCD endosomes. DHT does not appear to elicit general proteolysis, however, in addition to AQP2 degradation, DHT treatment also diminished micro calpain and calpastatin expression although whether these changes contributed to the AQP2 instability remains unclear. Together, these data show for the first time that AQP2 is a substrate for calpain-mediated proteolysis and that furthermore, micro-calpain, like AQP2, is both highly expressed in renal inner medulla and localized to apical IMCD endosomes. PMID- 12646167 TI - Reinvestigation on the causes of genomic GC variation between the orthologous genes of Mycobacterium tuberculosis and Mycobacterium leprae. AB - Genomic GC (overall G+C content of the coding sequences) variations were reinvestigated between the orthologous genes of Mycobacterium tuberculosis and Mycobacterium leprae species. It was observed that overall genomic GC variation between the species mainly originates from the combined effects GC(1) and GC(2) variations. But codons having identical amino acids with different codons (IA) (between the orthologous codon pairs) are responsible for the genomic GC(3) variation between the organisms, whereas orthologous codons having different amino acids (DA) between the two organisms are responsible for the variation of GC(1) levels. Further analyses indicate that duets and quartets are going in the same direction with same magnitude in changing the GC(3) levels for IA category, whereas GC(1) levels of duets of DA category decreases significantly from the overall GC(1) levels but GC(1) levels of quartets increases significantly from the overall GC(1) levels. GC(3) levels of informational genes for the IA category decrease more rapidly than the other functional categories of genes. The biological implications of these results have been discussed in this paper. PMID- 12646169 TI - LPS regulate ERK1/2-dependent signaling in cardiac fibroblasts via PKC-mediated MKP-1 induction. AB - Activation of MAPK pathways by angiotensin II (Ang II) is important for cardiac fibroblast (CFB) proliferation and migration. Activity of MAP-kinases is closely controlled by a group of dual-specific MAP kinase phosphatases (MKPs). Lipopolysaccharides (LPS) and cytokines are elevated in patients with heart failure and may contribute to disease progression. In this study, we investigate the effect of LPS on Ang II-induced CFB function. Pretreatment of CFBs with LPS (1 microg/mL; 30 min) almost completely inhibited Ang II-induced DNA-synthesis and inhibited Ang II directed chemotaxis by more than 80%. Compared to controls, LPS pretreatment significantly reduced phosphorylation levels of ERK1/2- and p38 MAPK and induced MKP-1 levels. Silencing MKP-1 with antisense oligodesoxynucleotides reversed the antimitogenic effect of LPS on Ang II-induced CFB DNA-synthesis and migration. Induction of MKP-1 by LPS was inhibited by the protein kinase C (PKC)-inhibitor calphostin C, but not by the ERK1/2-pathway inhibitor PD98059, suggesting that PKC but not ERK1/2 is required for LPS mediated MKP-1 induction in CFBs. Our data demonstrate that LPS have direct cellular effects in CFBs through an inhibition of Ang II-induced MAPK activity via PKC-mediated induction of MKP-1. This might be relevant with regard to the decreased MAPK activity and increased levels in MKPs reported during chronic heart failure in humans. PMID- 12646168 TI - ASC is an activating adaptor for NF-kappa B and caspase-8-dependent apoptosis. AB - ASC is a pro-apoptotic protein containing a pyrin domain (PD) and a caspase recruitment domain (CARD). A previous study suggests that ASC interacts with Ipaf, a member of the Apaf-1/Nod1 protein family. However, the functional relevance of the interaction has not been determined. Here, we report that co expression of ASC with Ipaf or oligomerization of ASC induces both apoptosis and NF-kappa B activation. Apoptosis induced through ASC was inhibited by a mutant form of Caspase-8 but not by that of Caspase-1. The PD of ASC physically interacted with Caspase-8 as well as with pyrin, the familial Mediterranean fever gene product. Caspase-8 deficiency rescued mouse fibroblasts from apoptosis induced by ASC oligomerization. Pyrin disrupted the interaction between ASC and Caspase-8, and inhibited both apoptosis and NF-kappa B activation induced by ASC. These findings suggest that ASC is a mediator of NF-kappa B activation and Caspase-8-dependent apoptosis in an Ipaf signaling pathway. PMID- 12646170 TI - NMR study of hexanucleotide d(CCGCGG)2 containing two triplet repeats of fragile X syndrome. AB - Long repeated stretches of d(CCG) and tri-nucleotide are crucial mutations that cause hereditary forms of mental retardation (fragile X-syndrome). Moreover, the alternating (CG) di-nucleotide is one of the candidates for Z-DNA conformation. Solution NMR structure of d(CCGCGG)(2) has been solved and is discussed. The determined NMR solution structure is a distorted highly bent B-DNA conformation with increased flexibility in both terminal residues. This conformation differs significantly from the Z-DNA tetramer structure reported for the same hexamer in the crystal state at similar ionic strength by Malinina and co-workers. Crystal structure of d(CCGCGG)(2) at high salt concentration includes a central alternating tetramer in Z-DNA conformation, while the initial cytosine swings out and forms a Watson-Crick base-pair with the terminal guanine of a symmetry related molecule. In solution, NMR data for sugar ring puckering combined with restrained molecular dynamics simulations starting from a Z-DNA form show that terminal furanose residues could adopt the conformation required for aromatic bases swinging out. Therefore, tetramer formation could be considered possible once the hexanucleotide had previously adopted the Z-DNA form. This work gives some insight into correlations between anomalous crystal structures and their accessibility in the solution state. PMID- 12646171 TI - A novel mammalian endoplasmic reticulum ubiquitin ligase homologous to the yeast Hrd1. AB - The yeast hHrd1 is a ubiquitin-protein ligase (E3) involved in ER-associated degradation. It was originally identified by genetic methods as an E3 of the yeast cholesterol biosynthetic enzyme HMG-CoA reductase (HMGR). We report the identification and cloning of a human homologue of Hrd1 (hHrd1). Immunofluorescence imaging confirms that the endogenous hHrd1 resides in the ER and in vitro assay demonstrates that it has a ubiquitin-ligase activity. However, the homology between the human and yeast Hrd1 is limited to the N-terminal domain of the proteins, and hHrd1 does not appear to be involved in the degradation of mammalian HMGR. PMID- 12646172 TI - Evidence for phosphorylation requirement for human bilirubin UDP glucuronosyltransferase (UGT1A1) activity. AB - Our discovery of rapid down-regulation of human bilirubin UDP glucuronosyltransferase (UGT) in colon cell lines that was transient and irreversible following curcumin- and calphostin-C-treatment, respectively, suggested phosphorylation event(s) were involved in activity. Likewise, bilirubin UGT1A1 expressed in COS-1 cells was inhibited by curcumin and calphostin-C. Because calphostin-C is a highly specific protein kinase C (PKC) inhibitor, we examined and found 4 to 5 predicted PKC phosphorylation sites in 11 UGTs examined. UGT1A1 incorporated [33P]orthophosphate, which was inhibited by calphostin-C. Also triple mutant, T75A/T112A/S435G-UGT1A1, at predicted PKC sites failed to incorporate [33P]orthophosphate. Individual or double mutants exhibited dominant-negative, additive, or no effect, while the triple mutant retained 10 15% activity towards bilirubin and two xenobiotics. Compared to wild-type, S435G and T112A/S435G shifted pH-optimum for eugenol, but not for bilirubin or anthraflavic acid, toward alkaline and acid conditions, respectively. This represents the first evidence that a UGT isozyme requires phosphorylation for activity. PMID- 12646173 TI - Gene expression of detoxifying enzymes in AhR and Nrf2 compound null mutant mouse. AB - The arylhydrocarbon receptor (AhR) regulates the expression of cytochrome P450 (CYP)-1 gene family members which catalyze xenobiotic Phase I metabolism, while Nrf2 exerts the concerted regulation of Phase II enzyme genes. We generated AhR and Nrf2 compound null mutant mice to examine the integrated function of AhR- and Nrf2-regulated enzymes in detoxification. Furthermore, we used this mouse model, by administering three different classes of chemical inducers, to examine how xenobiotic metabolism may be influenced in the absence of signals transduced by AhR or Nrf2. The compound mutant mice responded only weakly to AhR ligand or Phase II inducer, while they displayed a clear response to phenobarbital, an inducer of the CYP2B family through another, unrelated transcription factor. Here, we report an initial characterization of the AhR-Nrf2 double mutant mice, which may serve as a simplified bioassay system to evaluate xenobiotic toxicity and metabolic biotransformation of various drugs and environmental chemicals. PMID- 12646175 TI - Regulation of orexin neurons by the monoaminergic and cholinergic systems. AB - Orexins are a pair of neuropeptides implicated in energy homeostasis and arousal. Here we characterize the electrophysiological properties of orexin neurons using slice preparations from transgenic mice in which orexin neurons specifically express green fluorescent protein. Orexin neurons showed high frequency firing with little adaptation by injecting a positive current. The hyperpolarization activated current was observed in orexin neurons by a negative current injection. The neurotransmitters, which were implicated in sleep/wake regulation, affected the activity of orexin neurons; noradrenaline and serotonin hyperpolarized, while carbachol depolarized orexin neurons in either the presence or absence of tetrodotoxin. It has been reported that orexins directly or indirectly activate the nuclei that are the origin of the neurons containing these neurotransmitters. Our data suggest that orexin neurons have reciprocal neural circuitries between these nuclei for either a positive or negative feedback loop and orchestrate the activity of these neurons to regulate the vigilance states. PMID- 12646174 TI - Gliclazide protects pancreatic beta-cells from damage by hydrogen peroxide. AB - Oxidative stress is induced under diabetic conditions and possibly causes various forms of tissue damage in patients with diabetes. Recently, it has become aware that susceptibility of pancreatic beta-cells to oxidative stress contributes to the progressive deterioration of beta-cell function in type 2 diabetes. A hypoglycemic sulfonylurea, gliclazide, is known to be a general free radical scavenger and its beneficial effects on diabetic complications have been documented. In the present study, we investigated whether gliclazide could protect pancreatic beta-cells from oxidative damage. One hundred and fifty microM hydrogen peroxide reduced viability of mouse MIN6 beta-cells to 29.3%. Addition of 2 microM gliclazide protected MIN6 cells from the cell death induced by H(2)O(2) to 55.9%. Glibenclamide, another widely used sulfonylurea, had no significant effects even at 10 microM. Nuclear chromatin staining analysis revealed that the preserved viability by gliclazide was due to inhibition of apoptosis. Hydrogen peroxide-induced expression of an anti-oxidative gene heme oxygenase-1 and stress genes A20 and p21(CIP1/WAF1), whose induction was suppressed by gliclazide. These results suggest that gliclazide reduces oxidative stress of beta-cells by H(2)O(2) probably due to its radical scavenging activity. Gliclazide may be effective in preventing beta-cells from the toxic action of reactive oxygen species in diabetes. PMID- 12646176 TI - Inhibition of human telomerase reverse transcriptase gene expression by BRCA1 in human ovarian cancer cells. AB - Human telomerase reverse transcriptase (hTERT), the catalytic subunit of human telomerase, is responsible for the synthesis and maintenance of the telomeric repeats at the distal ends of human chromosomes. Telomerase expression is repressed in normal human cells and is activated in immortal cells and during tumorigenesis, but the mechanism by which telomerase expression is regulated is not fully understood. Previous studies have shown that c-Myc stimulates hTERT transcription through the binding sites located on the hTERT promoter. In this study, we sought to determine whether BRCA1 inhibits hTERT transcription through its direct interaction with c-Myc. In ovarian cancer cells, c-Myc increased hTERT expression by threefold and BRCA1 completely abrogated this activity. A mutation in the c-Myc-binding site (E-box) of the hTERT promoter resulted in the loss of activation by c-Myc and in the loss of inhibition by BRCA1. Deletion of the c-Myc binding domain in BRCA1 resulted in the loss of BRCA1's ability to inhibit transcription of the hTERT promoter. In addition, BRCA1 associates with c-Myc and inhibits the binding activity of c-Myc to the hTERT promoter. Our data indicate that BRCA1 is involved in regulating cellular immortalization through the modulation of c-Myc on the hTERT promoter. PMID- 12646177 TI - A role of the Duffy antigen for the maintenance of plasma chemokine concentrations. AB - We examined plasma chemokine concentrations and chemokine clearance rates in Duffy antigen knockout mice. The plasma concentrations of eotaxin and MCP-1 in Duffy antigen knockout mice were less than one-third of those in wild-type mice. When eotaxin or hMGSA was intravenously injected, the chemokine disappeared more rapidly from the plasma of Duffy antigen knockout mice than from the plasma of wild-type mice. The half-lives of hIP-10 and interferon-gamma, which do not have an affinity for the Duffy antigen, in plasma were indistinguishable between Duffy antigen knockout mice and wild-type mice. These results suggest that the Duffy antigen delays the disappearance of chemokines from the plasma, resulting in the maintenance of plasma chemokine concentrations. PMID- 12646178 TI - Regulation of YB-1 gene expression by GATA transcription factors. AB - GATA-1 is a transcription factor essential for erythroid cell development, and knockdown of GATA-1 gene results in maturation arrest and transformation of erythroblasts. To clarify the mechanism that gives rise to this abnormal phenotype, genes that are aberrantly expressed in the spleen of heterozygous GATA 1 knockdown mutant mouse (referred to as GATA-1 mutant mouse) were identified by using cDNA array. One of these genes, YB-1, was found to be highly expressed in the spleen of GATA-1 mutant mouse. Reporter and electrophoretic mobility shift assays revealed that the proximal GATA element in 5'-UTR region of YB-1 gene functions positively in K562 cells. Furthermore, both GATA-1 and GATA-2, which were transiently expressed in COS-7 cells, bound to this element and activated the YB-1 promoter through this element. These results suggest that YB-1 functions under the regulation of GATA factors in erythroid differentiation and aberrant expression of YB-1 gene may result in dyserythropoiesis. PMID- 12646179 TI - Molecular identification of a Drosophila G protein-coupled receptor specific for crustacean cardioactive peptide. AB - The Drosophila Genome Project website (www.flybase.org) contains the sequence of an annotated gene (CG6111) expected to code for a G protein-coupled receptor. We have cloned this receptor and found that its gene was not correctly predicted, because an annotated neighbouring gene (CG14547) was also part of the receptor gene. DNA corresponding to the corrected gene CG6111 was expressed in Chinese hamster ovary cells, where it was found to code for a receptor that could be activated by low concentrations of crustacean cardioactive peptide, which is a neuropeptide also known to occur in Drosophila and other insects (EC(50), 5.4 x 10(-10)M). Other known Drosophila neuropeptides, such as adipokinetic hormone, did not activate the receptor. The receptor is expressed in all developmental stages from Drosophila, but only very weakly in larvae. In adult flies, the receptor is mainly expressed in the head. Furthermore, we identified a gene sequence in the genomic database from the malaria mosquito Anopheles gambiae that very likely codes for a crustacean cardioactive peptide receptor. PMID- 12646180 TI - Atomic-resolution STM structure of DNA and localization of the retinoic acid binding site. AB - Single-molecule imaging by scanning tunnelling microscopy (STM) yields the atomic resolution (0.6A) structure of individual B-type DNA molecules. The strong correlation between these STM structures and those predicted from the known base sequence indicates that sequencing of single DNA molecules using STM may be feasible. There is excellent agreement between the STM and X-ray structures, but subtle differences exist due to radial distortions. We show that the interactions of other molecules with DNA, their binding configurations, and the structure of these complexes can be studied at the single-molecule level. The anti-cancer drug retinoic acid (RA) binds selectively to the minor groove of DNA with up to 6 RA molecules per DNA turn and with the plane of the RA molecule approximately parallel to the DNA symmetry axis. Similar studies for other drug molecules will be valuable in the a priori evaluation of the effectiveness of anti-cancer drugs. PMID- 12646181 TI - Characterization of alternatively spliced and truncated forms of the Arf guanine nucleotide exchange factor GBF1 defines regions important for activity. AB - Analysis of multiple transcripts for the Arf-specific guanine nucleotide exchange factor GBF1 identified three positions displaying small in-frame deletions and insertions. Sequencing of genomic DNA for CHO GBF1 and analysis of the human gene established that those variations were consistent with alternate splicing events. RT-PCR analysis of CHO mRNA confirmed that these small in-frame deletions occurred at significant and similar frequencies in both WT and BFA resistant CHO cells. These splice variants behaved like GBF1 in biological assays based on the observation that GBF1 is cytotoxic at high levels but will confer resistance to BFA when moderately overexpressed. Comparison of variants with larger deletions defined regions of 75 (exons 5-7) and 412 (exons 31-39) amino acid residues that were required for cell killing but were dispensable for promoting BFA resistance. PMID- 12646183 TI - Association of the low-density lipoprotein receptor with caveolae in hamster and rat liver. AB - The association of the low-density lipoprotein (LDL) receptor with detergent resistant hepatic membranes was investigated using discontinuous sucrose gradients. In liver homogenates from both hamsters and rats, the fractions with the highest concentrations of LDL receptor coincided with the location of caveolin-1, a marker of the cholesterol-rich caveolae. Feeding the animals diets enriched in cholesterol slightly shifted both LDL receptor and caveolin-1 to positions of lower density. The cholesterol content of the caveolae fractions was increased 2-fold in animals fed cholesterol-supplemented diets. In homogenates of CHO cells, fractionated in the same manner, the LDL receptor was absent from the caveolae fractions but was present in denser fractions near the bottom of the gradient. Addition of caveolin-1 antibody to solubilized caveolae from liver coimmunoprecipitated the LDL receptor. These observations suggest that in liver, the LDL receptor is mainly located in caveolae. This location contrasts with the clathrin-coated pit location observed in fibroblasts and CHO cells. PMID- 12646182 TI - New selective amplifier genes containing c-Mpl for hematopoietic cell expansion. AB - We previously developed "selective amplifier genes (SAGs)" which confer a growth advantage to transduced cells. The SAG is a chimeric gene encoding the G-CSF receptor (GCR) and the estrogen or tamoxifen (Tm) receptor and is able to expand transduced hematopoietic cells by treatment with estrogen or Tm. In the current study, we examined the in vitro efficacy of modified SAGs containing the thrombopoietin (TPO) receptor (c-Mpl) gene instead of GCR as a more potent signal generator. In addition, we constructed various mutant Mpl-type SAGs to abolish the responsiveness to endogenous TPO while retaining Tm-dependency. When Ba/F3 cells were retrovirally transduced with the Mpl-type SAGs, the cells showed Tm- and TPO-dependent growth even without IL-3. The Mpl-type SAGs induced more potent proliferation of Ba/F3 and cynomolgus CD34(+) cells than the GCR-type SAG. One mutant Mpl-type SAG (Delta GCRMplTmR) successfully lost the responsiveness to TPO without affecting the Tm-dependence. PMID- 12646184 TI - Characterization of boar sperm cytoskeletal cylicin II as an actin-binding protein. AB - The presence of actin-binding proteins in the perinuclear theca of boar spermatozoa has been investigated, using stepwise extractions of proteins from sperm heads. Proteins extracted with the alkaline buffer 1M Na(2)CO(3), pH 11, were found to contain a 66kDa protein that binds F-actin in actin pelleting assays. Sequence studies and immunological characterization with antibodies specific for human cylicin II identified the 66kDa protein as the homologue of bovine and human cylicin II. Immunocytochemical studies showed the presence of porcine cylicin II in the acrosomal region of round spermatids and in the postacrosomal region of late spermatids and spermatozoa, in agreement with the previously described localization of cylicins. Taken together, the results suggest that cylicin II, a protein of the sperm perinuclear cytoskeleton, is a novel actin-binding protein, which probably plays a role in the actin-related events that occur during spermiogenesis and the early events of fertilization. PMID- 12646185 TI - ETS2 is involved in protein kinase C-activated expression of granulocyte macrophage colony-stimulating factor in human non-small lung carcinoma cell line, A549. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine expressed in the non-small lung carcinoma cells (NSCLC). However, transcriptional regulation of GM-CSF is not well characterized in NSCLC. In this study we found that two cis-acting ETS family consensus sites are important for transcriptional regulation of GM-CSF in A549 human lung carcinoma cells. These two sites are located separately at around -40 and -100 bp from the transcription start site. Results of transient transfection assays with A549 cells indicated that ETS2 had a strong positive effect on GM-CSF promoter activity. Furthermore, this activity was enhanced by protein kinase C activator, phorbol 12-myristate 13-acetate (PMA), in an ETS consensus-dependent manner, while PMA could also enhance the expression level of ETS2. The protein kinase C inhibitors decreased GM-CSF promoter activity induced by the protein kinase C activator PMA. We also found that antisense ETS2 mRNA decreased PMA-induced GM-CSF promoter activity, supporting the possibility that ETS2 is involved in protein kinase C-induced GM CSF transcriptional function. Endogenous expression of GM-CSF mRNA was increased by ETS2 transfection and the increased expression was further enhanced by PMA. These data indicate that GM-CSF is up-regulated by ETS2, a target of protein kinase C. PMID- 12646186 TI - Insights into the regulation of heat shock transcription factor 1 SUMO-1 modification. AB - The transcriptional regulatory protein HSF1 is the key mediator of induced heat shock protein gene expression in response to elevated temperature and other stresses. Our previous studies identified stress-induced SUMO-1 modification of HSF1 as an important regulator of the DNA-binding activity of this factor. The underlying molecular mechanism by which stress leads to sumoylation of HSF1 was unknown. Prompted by previous studies indicating stress-induced phosphorylation at serine 307 of HSF1, a site very near the sumoylation site at lysine 298, we examined the role of this phosphorylation event in regulating SUMO-1 modification of HSF1. Using a combination of transfection and in vitro phosphorylation/sumoylation experiments, our results indicate that phosphorylation at serine 307 stimulates sumoylation of HSF1. Our results also reveal a role for a conserved leucine zipper sequence in the C-terminal region of HSF1 in inhibiting its SUMO-1 modification. Based on these data, we postulate that phosphorylation at serine 307 could stimulate HSF1 sumoylation by causing a conformation change that relieves the inhibitory effect of the C-terminal leucine zipper. PMID- 12646187 TI - Efficient preparation of glycoprotein hormones lacking an alpha-subunit oligosaccharide. AB - The oligosaccharide on alpha-subunit loop 2 (alpha 2) is needed for full glycoprotein hormone efficacy. Efforts to prepare glycoprotein hormone antagonists usually involve removing the alpha 2 oligosaccharide and are hampered by its requirement for efficient heterodimer secretion from mammalian cells. Here we show that hormones lacking this oligosaccharide can be produced by treating them at low pH to dissociate the heterodimer and permitting the subunits to re associate in the presence of peptide N-glycosidase F (PNGase F). Re-assembly of human choriogonadotropin, human follitropin, and bovine lutropin occurred rapidly and efficiently following removal of the alpha 2 oligosaccharide by PNGase F. Consequently, virtually all heterodimers formed in the presence of this enzyme lacked this oligosaccharide. These findings support the notion that heterodimer assembly in vitro occurs by a threading mechanism that is impeded by the presence of the alpha 2 oligosaccharide. This procedure should facilitate the study of glycoprotein hormone structure and function. PMID- 12646188 TI - Protein kinase C alpha and zeta regulate nitric oxide-induced NF-kappa B activation that mediates cyclooxygenase-2 expression and apoptosis but not dedifferentiation in articular chondrocytes. AB - Nitric oxide (NO) regulates differentiation, survival, and cyclooxygenase (COX)-2 expression in articular chondrocytes. NO-induced apoptosis and dedifferentiation are mediated by p38 kinase activity and p38 kinase-independent and -dependent inhibition of protein kinase C (PKC)alpha and zeta. Because p38 kinase also activates NF-kappa B, we investigated the functional relationship between PKC and NF-kappa B signaling and the role of NF-kappa B in apoptosis, dedifferentiation, and COX-2 expression. We found that NO-stimulated NF-kappa B activation was inhibited by ectopic PKC alpha and zeta expression, whereas NO-stimulated inhibition of PKC alpha and zeta activity was not affected by NF-kappa B inhibition. Inhibition of NO-induced NF-kappa B activity did not affect inhibition of type II collagen expression but did abrogate COX-2 expression and apoptosis. Taken together, our results indicate that NO-induced inhibition of PKC alpha and zeta activity is required for the NF-kappa B activity that regulates apoptosis and COX-2 expression but not dedifferentiation in articular chondrocytes. PMID- 12646189 TI - PECAM-1 promotes beta-catenin accumulation and stimulates endothelial cell proliferation. AB - Platelet endothelial cell adhesion molecule-1 (PECAM-1) binds tyrosine phosphorylated beta-catenin and modulates beta-catenin localization [J. Immunol. 158 (7) (1997) 3408; J. Cell Sci. 112 (Pt 18) (1999) 3005]. To elucidate functional consequences of this interaction, we studied endothelial cells from PECAM-1 knockout animals and compared them to PECAM-1 expressing endothelial cells [Mol. Biol. Cell 11 (9) (2000) 3109]. We noted an increase in the expression of beta-catenin protein in PECAM-1 expressing endothelial cells. Further, by immunofluorescence, beta-catenin localized to the cell membrane as well as to the nucleus in PECAM-1 positive endothelial cells, whereas cells not expressing PECAM-1 stained for beta-catenin only at the membrane. Additionally, we demonstrate that PECAM-1 lacking the majority of the cytoplasmic domain promotes significantly less accumulation of transcriptionally active beta-catenin than full-length PECAM-1. Finally, we note an increased proliferative rate in the PECAM-1 reconstituted cells compared to the endothelial cells lacking PECAM-1. Taken together, our data suggest that PECAM-1, an adhesion molecule, affects cell proliferation via accumulation of transcriptionally active beta-catenin. PMID- 12646191 TI - Characterization of aldehyde ferredoxin oxidoreductase gene defective mutant in Magnetospirillum magneticum AMB-1. AB - A non-magnetic mutant of Magnetospirillum magneticum AMB-1, designated as NMA21, was generated by mini-Tn5 transposon mutagenesis to identify genes involved in bacterial magnetic particle (BMP) synthesis. Alignment of the DNA sequences flanking the transposon allowed the isolation of an open reading frame (ORF2) within an operon consisting of five genes. The amino acid sequence of ORF2 showed homology with tungsten-containing aldehyde ferredoxin oxidoreductase (AOR) from Pyrococcus furiosus (48% identity and 64% similarity), which functions for aldehyde oxidation. AOR was found to be expressed under microaerobic conditions and localized in the cytoplasm of AMB-1. Iron uptake and growth of NMA21 were lower than wild type. Transmission electron microscopy (TEM) of NMA21 revealed that no BMPs were completely synthesized, but polyhydroxybutyrate (PHB)-like granules were persistently produced. These results indicate that AOR may contribute to ferric iron reduction during BMP synthesis in M. magneticum AMB-1 under microaerobic respiration. PMID- 12646190 TI - P202, an interferon-inducible protein, inhibits E2F1-mediated apoptosis in prostate cancer cells. AB - p202, an interferon (IFN) inducible protein, is a phosphonuclear protein involved in the regulation of cell cycle, apoptosis, and differentiation. E2F1 belongs to the E2F family of proteins that are important cell cycle regulators in promoting cell growth. On the other hand, the deregulated expression of E2F1 also triggers apoptosis independent of p53 status. It has been well documented that p202 is able to inhibit cell growth by binding to E2F1 and abolishing the E2F1-mediated transcriptional activation of S-phase genes. However, it is not known whether E2F1-mediated apoptosis can be counteracted by p202 expression. Here, we show that E2F1-mediated apoptosis induced by the infection of an E2F1-expressing adenoviral vector (Ad-E2F1) was greatly diminished in p202-expressing prostate cancer cells. The E2F1-mediated caspase-3 activation was also reduced in p202 expressing cells infected with Ad-E2F1. Since caspase-3 is one of the E2F1 transcriptional targets, this result is consistent with the ability of p202 to inhibit the transcriptional activity of E2F1. Therefore, our results suggest a possible link between the IFN and E2F pathways in regulating apoptosis. PMID- 12646192 TI - Tissue-specific modulation of beta-adducin transcripts in Milan hypertensive rats. AB - Genetic variants in Adducins, a family of cytoskeleton proteins (alpha, beta, and gamma) encoded by three genes, have been associated with primary hypertension in humans and in Milan hypertensive (MHS) rats. The present paper describes the identification of a rat beta 4 alternative splicing isoform differing from beta subunit for an in-frame insertion of 18 amino acids and showing a polymorphic site (R592W) between MHS and its normotensive control (MNS). Furthermore, we established a quantitative real-time PCR assay for analyzing the tissue expression of adducin gene family and determining whether any subunit transcript demonstrates altered expression during the development of MHS hypertension, especially in tissues relevant for the control of cardiovascular phenotypes (i.e., kidney, left ventricle, and large arteries). Among the three adducins only beta transcripts were modulated, in a tissue-specific manner, during the development of hypertension in MHS, compared to age-matched MNS controls. A 43% decrease in renal outer medulla was already present at the prehypertensive phase; a 70% decrease in femoral artery and 66% increase in left ventricle were observed after the development of hypertension. Surprisingly beta 4-Add, which is a minor component of total beta transcripts, is drastically reduced up to 88% in all MHS tissues. Alteration in beta-Add expression levels may account, at least in part, for the observed phenotypic changes in MHS hypertension. PMID- 12646194 TI - Oxidized LDL receptor gene (OLR1) is associated with the risk of myocardial infarction. AB - Lectin-like oxidized low-density lipoprotein receptor (LOX-1/OLR1) has been suggested to play a role in the progression of atherogenesis. We analyzed the OLR1 gene and found a single nucleotide polymorphism (SNP), G501C, in patients with ischemic heart disease from a single family, which resulted in the missense mutation of K167N in LOX-1 protein. We compared the group of patients with myocardial infarction (MI) (n=102) with a group of clinically healthy subjects (n=102), and found that the MI group had a significantly high frequency of 501G/C+501C/C (38.2%) compared with the healthy group (17.6%; p<0.002). The odds ratio for the risk of MI associated with the 501G/C+501C/C genotype was 2.89 (95% CI, 1.51-5.53). These findings suggest that OLR1 or a neighboring gene linked with G501C SNP is important for the incidence of MI. Manipulating LOX-1 activity might be a useful therapeutic and preventative approach for coronary artery disease, especially for individuals with the G501C genotype of OLR1. PMID- 12646193 TI - Contryphan-Vn: a modulator of Ca2+-dependent K+ channels. AB - Contryphan-Vn is a D-tryptophan-containing disulfide-constrained nonapeptide isolated from the venom of Conus ventricosus, the single Mediterranean cone snail species. The structure of the synthetic Contryphan-Vn has been determined by NMR spectroscopy. Unique among Contryphans, Contryphan-Vn displays the peculiar presence of a Lys-Trp dyad, reminiscent of that observed in several voltage-gated K(+) channel blockers. Electrophysiological experiments carried out on dorsal unpaired median neurons isolated from the cockroach (Periplaneta americana) nerve cord on rat fetal chromaffin cells indicate that Contryphan-Vn affects both voltage-gated and Ca(2+)-dependent K(+) channel activities, with composite and diversified effects in invertebrate and vertebrate systems. Voltage-gated and Ca(2+)-dependent K(+) channels represent the first functional target identified for a conopeptide of the Contryphan family. Furthermore, Contryphan-Vn is the first conopeptide known to modulate the activity of Ca(2+)-dependent K(+) channels. PMID- 12646195 TI - Rapamycin inhibits E2F-dependent expression of minichromosome maintenance proteins in vascular smooth muscle cells. AB - Rapamycin inhibits vascular smooth muscle cell (VSMC) proliferation and rapamycin eluting stents represent a novel therapeutic strategy for preventing postangioplasty restenosis. The precise molecular mechanism, for rapamycin mediated inhibition of VSMC cell cycle progression and DNA replication remain to be elucidated. Minichromosome maintenance proteins (MCM) are essential regulators of DNA replication and the objective of this study was to examine the effect of rapamycin on their expression in rat aortic VSMC. Rapamycin substantially inhibited mitogen-induced MCM6 and MCM7 mRNA and protein expression in a dose dependent fashion. Transient transfection experiments revealed that rapamycin inhibited MCM6 and MCM7 promoter activity, implicating a transcriptional mechanism. MCM6 and MCM7 transcriptional activation is regulated by E2F and activity of a luciferase reporter plasmid driven by four E2F elements was also significantly inhibited by rapamycin. The inhibitory effect of rapamycin on MCM6 and MCM7 was reversed by overexpression of E2F, indicating that their downregulation by rapamycin involves an E2F-dependent mechanism. These observations suggest that rapamycin inhibits MCM6 and MCM7 expression by blocking their E2F-dependent transactivation which may contribute importantly to the inhibition of VSMC DNA synthesis by rapamycin. PMID- 12646196 TI - Effects of maturation on RNA transcription and protein expression of four MRP genes in human placenta and in BeWo cells. AB - The placenta is a multifunctional organ that protects the fetus from toxic compounds and the MRPs contribute to this function. The expression of MRP1, MRP2, MRP3, and MRP5 was compared in human placental tissue and in BeWo cells by real time RT-PCR analysis; protein expression was assessed by Western blot. MRP1 and MRP3 were the most abundantly expressed genes in placenta but only MRP1 was highly expressed in the BeWo cells. Expression of MRP1 increased 4-fold in the third as compared with first trimester placental samples, and increased 20-fold with polarization of BeWo cells. MRP2, MRP3, and MRP5 were weakly expressed both in placenta and BeWo cells. Protein expression followed mRNA quantification for MRP1 and MRP5 but not for MRP2 and MRP3. These data indicated that MRP1 and MRP5 increase with trophoblast maturation, suggesting a particular role for these proteins in the organ functional development. PMID- 12646197 TI - The immunosuppressive agent tacrolimus induces p21WAF/CIP1WAF1/CIP1 via TGF-beta secretion. AB - Tacrolimus (Tac) is more immunosuppressive drug compared to cyclosporine (CsA). Our previous studies have demonstrated that CsA induces the expression of p21WAF/CIP1 expression. In this study we explored if like CsA, Tac also induces expression of p21WAF/CIP1. We also determined if induction of p21WAF/CIP1 by Tac is dependent on TGF-beta. Using RT-PCR and Western blot analysis, we studied the induction of p21WAF/CIP1 mRNA and protein in human T cells and A-549 cells (human lung adenocarcinoma cells) by Tac. The stimulation of p21WAF/CIP1 promoter activity was studied by luciferase assay using p21WAF/CIP1-luc, chimeric plasmid DNA containing a p21WAF/CIP1 promoter segment and luciferase reporter gene. Using anti-TGF-beta antibody, we studied if induction of p21WAF/CIP1 by tacrolimus is dependent on TGF-beta. The results demonstrate that Tac induced p21WAF/CIP1 mRNA and protein expression as well as stimulated its promoter activity in T cells and A-549 cells. The induction of p21WAF/CIP1 expression by tacrolimus was dependent on TGF-beta since a neutralizing anti-TGF-beta antibody inhibited induction of p21WAF/CIP1in A-549 cells. These data support the hypothesis that cyclin inhibitor p21WAF/CIP1 might represent a unified mediator of the anti proliferative effects of Tac and other immunosuppressive agents. Strategies involving p21WAF/CIP1 induction should be considered a viable alternative strategy to achieve immunosuppression possibly with reduced toxicity associated with current immunosuppression. PMID- 12646198 TI - 13-cis-retinoic acid competitively inhibits 3 alpha-hydroxysteroid oxidation by retinol dehydrogenase RoDH-4: a mechanism for its anti-androgenic effects in sebaceous glands? AB - Retinol dehydrogenase-4 (RoDH-4) converts retinol and 13-cis-retinol to corresponding aldehydes in human liver and skin in the presence of NAD(+). RoDH-4 also converts 3 alpha-androstanediol and androsterone into dihydrotestosterone and androstanedione, which may stimulate sebum secretion. This oxidative 3 alpha hydroxysteroid dehydrogenase (3 alpha-HSD) activity of RoDH-4 is competitively inhibited by retinol and 13-cis-retinol. Here, we further examine the substrate specificity of RoDH-4 and the inhibition of its 3 alpha-HSD activity by retinoids. Recombinant RoDH-4 oxidized 3,4-didehydroretinol-a major form of vitamin A in the skin-to its corresponding aldehyde. 13-cis-retinoic acid (isotretinoin), 3,4-didehydroretinoic acid, and 3,4-didehydroretinol, but not all trans-retinoic acid or the synthetic retinoids acitretin and adapalene, were potent competitive inhibitors of the oxidative 3 alpha-HSD activity of RoDH-4, i.e., reduced the formation of dihydrotestosterone and androstandione in vitro. Extrapolated to the in vivo situation, this effect might explain the unique sebosuppressive effect of isotretinoin when treating acne. PMID- 12646199 TI - Dibenzoylmethane, a natural dietary compound, induces HIF-1 alpha and increases expression of VEGF. AB - Hypoxia-inducible factor 1 (HIF-1) is the major transcription factor activated during hypoxia. It is composed of HIF-1 alpha and HIF-1 beta subunits. While HIF 1 beta is constitutively expressed, HIF-1 alpha is targeted to proteasome degradation under normoxic conditions. Under hypoxia, HIF-1 alpha is stabilized and heterodimerizes with HIF-1 beta. Iron chelators have also been reported to stabilize HIF-1 alpha protein and activate HIF-1. In this study, we investigated the effects of dibenzoylmethane (DBM), a natural dietary compound and an iron chelator, on HIF-1 pathway. We found that DBM increases HIF-1 alpha protein levels in a dose- and time-dependent manner. This induction was accompanied with activation of HIF-1, measured by reporter gene assay and increased production of its downstream target, the vascular endothelial growth factor. Mechanistically, HIF-1 alpha was stabilized by DBM at a step prior to ubiquitination. The effect of DBM on HIF-1 and its low toxicity profile might be therapeutically beneficial in ischemic diseases. PMID- 12646200 TI - Bio-effectiveness of Tat-catalase conjugate: a potential tool for the identification of H2O2-dependent cellular signal transduction pathways. AB - Reactive oxygen species such as hydrogen peroxide (H(2)O(2)) have taken center stage as bona fide second messengers in various signaling pathways. Here, we report the synthesis, metabolic fate, and effectiveness in modulating such pathways of a Tat-catalase conjugate. Incubation of L2 cells with Tat-catalase greatly increased cell-associated enzymatic activity, reaching close to a plateau by 30 min. The cell-associated catalase activity and antibody-detectable Tat derivatives declined over time after changing medium, although still remaining at significantly higher levels than baseline even at 4h. While most cell-associated Tat-catalase was apparently tightly attached to the cell surface, a small fraction entered the cells as the proteasome inhibitor MG-132 slightly prevented the disappearance of the enzyme. Tat-catalase, either membrane-bound or intracellular, but not native catalase, inhibited serum-induced Elk phosphorylation and anisomycin- and/or MG-132-induced ERK phosphorylation, suggesting the involvement of H(2)O(2). Thus, Tat-catalase should be a useful tool to dissect H(2)O(2)-dependent events in signaling pathways. PMID- 12646201 TI - Altered functionality in rhodopsin point mutants associated with retinitis pigmentosa. AB - Point mutations found in rhodopsin associated with the retinal degenerative disease retinitis pigmentosa have been expressed in mammalian COS-1 cells, purified, and characterised. The mutations characterised-most of them for the first time-have been Met44Thr, Gly114Asp, Arg135Leu, Val137Met, and Pro171Leu in the transmembrane domain; Leu328Pro and Ala346Pro in the C-terminal tail of the cytoplasmic domain; and Gly106Trp in the intradiscal domain. Several of these mutations cause misfolding which results in impaired 11-cis-retinal binding. Two of them, Met44Thr and Val137Met, show spectral and structural features similar to those of wild type rhodopsin (Type I mutants) but significantly increased transducin initial activation rates. We propose that, in the case of these mutants, abnormal functioning resulting in faster activation kinetics could also play a role in retinitis pigmentosa by altering the stoichiometric balance of the different proteins involved in the phototransduction biochemical reactions. PMID- 12646202 TI - beta-Amyloid-specific upregulation of stearoyl coenzyme A desaturase-1 in macrophages. AB - beta-Amyloid peptide (A beta), a major component of senile plaques, the formation of which is characteristic of Alzheimer's disease (AD), is believed to induce inflammation of the brain mediated by microglia, leading to neuronal cell loss. In this study, we performed an oligonucleotide microarray analysis to investigate the molecular events underlying the A beta-induced activation of macrophages and its specific suppression by the A beta-specific-macrophage-activation inhibitor, RS-1178. Of the approximately 36,000 genes and expressed sequence tags analyzed, eight genes were specifically and significantly upregulated by a treatment with interferon gamma (IFN gamma) and A beta compared to a treatment with IFN gamma alone (p<0.002). We found that the gene for a well-characterized lipogenetic enzyme, stearoyl coenzyme A desaturase-1 (SCD-1), was specifically upregulated by A beta treatment and was suppressed to basal levels by RS-1178. Although the underlying mechanisms remain unknown, our results suggest the presence of a link between AD and SCD-1. PMID- 12646203 TI - Temporal gene expression changes during adipogenesis in human mesenchymal stem cells. AB - Human bone marrow mesenchymal stem cells (hMSCs) give rise to adipocytes in response to adipogenic hormones. An in-house cDNA microarray representing 3400 genes was employed to characterize the modulation of genes involved in this process. A total of 197 genes showed temporal gene expression changes during adipogenesis, including genes encoding transcriptional regulators and signaling molecules. Semi-quantitative RT-PCR analyses confirmed differential expression at the transcriptional level of several genes identified by cDNA microarray screening. Cluster analysis of the genes regulated during the late phase (from day 7 to day 14) of hMSC adipogenesis indicated that these changes are well correlated with data previously reported for murine preadipocytes. However, during the early phase (day 1-day 5), the modulations of genes differed from those reported for the preadipocytes. These data provide novel information on the molecular mechanisms required for lineage commitment and maturation accompanying adipogenesis of hMSC. PMID- 12646204 TI - NF-kappa B activation in endothelial cells treated with oxidized high-density lipoprotein. AB - We first determined whether oxidized high-density lipoprotein (ox-HDL) activates transcription factor nuclear factor-kappa B (NF-kappa B) in cultured human umbilical vein endothelial cells (HUVECs). Treatment for 7h with 100 microg/ml ox HDL elicited a marked downregulation of I kappa B alpha and upregulation of the phosphorylated form of I kappa B alpha in HUVECs in a manner dependent on the dose of ox-HDL. Electrophoretic mobility shift assay in nuclear fraction from HUVECs showed translocation of NF-kappa B to the nucleus and binding of NF-kappa B to NF-kappa B consensus oligonucleotides during ox-HDL exposure for 7h, suggesting that ox-HDL brings about NF-kappa B activation in endothelial cells. To clarify the mechanism of NF-kappa B activation in HUVECs treated with ox-HDL, we investigated the effect of ox-HDL treatment on intracellular production of reactive oxygen species (ROS) in HUVECs. Ox-HDL induced a significant dose dependent increase in ROS production during 4h incubation and this enhanced production of ROS was inhibited in the presence of probucol or diphenylene iodonium (DPI), an inhibitor of NADPH oxidase. In addition, pretreatment with probucol or DPI suppressed the phosphorylation and degradation of I kappa B alpha protein induced by ox-HDL, demonstrating that increased generation of ROS by ox HDL may be associated with NF-kappa B activation. Pretreatment with antibody against oxidized low-density lipoprotein receptor-1 (LOX-1) significantly suppressed the ox-HDL-induced downregulation of I kappa B alpha, suggesting that LOX-1 mediates NF-kappa B activation in endothelial cells stimulated with ox-HDL. Taking all of the above findings together, ox-HDL activates NF-kappa B via binding to LOX-1 on the cell surface, followed by enhancement of intracellular ROS production in endothelial cells. PMID- 12646205 TI - Mouse mast cells express the tryptic protease neuropsin/Prss19. AB - The only tryptic enzymes identified so far in mouse mast cells (MCs) are three members of the chromosome 17A3.3 family of neutral proteases. Sequence analysis of a cDNA library revealed that BALB/c mouse bone marrow-derived MCs express neuropsin, a member of the chromosome 7B2 family of tryptic kallikreins. Kinetic studies revealed that neuropsin is expressed relatively early in MC development. As assessed immunohistochemically, the MCs residing in numerous connective tissues store neuropsin in their secretory granules. The finding that the neuropsin transcript is maximally expressed in the intestine at the height of a helminth infection indicates that MC-committed progenitors selectively increase their expression of neuropsin as they develop into mature mucosal MCs. This is the first report documenting the expression of neuropsin in an immune cell. Thus, it is now apparent that mouse MCs store at least two distinct families of tryptic like proteases in their secretory granules. PMID- 12646206 TI - An overexpression of chalcone reductase of Pueraria montana var. lobata alters biosynthesis of anthocyanin and 5'-deoxyflavonoids in transgenic tobacco. AB - We isolated the chalcone reductase (pl-chr) gene of Pueraria montana var. lobata by using a PCR strategy from cDNA pools of storage roots. A high level of expression of RNA was found in both stems and roots. The genomic Southern blot result suggests that pl-chr exists as a member of a small gene family. By introducing a pl-chr gene under the control of the 35S CaMV promoter into the pink-flowering Xanthi line of Nicotiana tabacum, the flower color was changed from pink to white-to-pink. The contents of anthocyanin in the flowers of the transgenic lines were dramatically decreased by 40%, but the total UV absorption compounds remained unchanged. The production of liquiritigenin in pl-chr overexpressed transgenic tobacco lines was confirmed by HPLC and MS analysis. The introduction of pl-chr gene provides a method to redirect the flavonoid pathway into 5'-deoxyflavonoid production in non-legume crops, in order to manipulate the phenylpropanoid pathway for isoflavonoid production. PMID- 12646207 TI - Na+/Mg2+ transporter acts as a Mg2+ buffering mechanism in PC12 cells. AB - Mg(2+) buffering mechanisms in PC12 cells were demonstrated with particular focus on the role of the Na(+)/Mg(2+) transporter by using a newly developed Mg(2+) indicator, KMG-20, and also a Na(+) indicator, Sodium Green. Carbonyl cyanide p (trifluoromethoxy) phenylhydrazone (FCCP), a protonophore, induced a transient increase in the intracellular Mg(2+) concentration ([Mg(2+)](i)). The rate of decrease of [Mg(2+)](i) was slower in a Na(+)-free extracellular medium, suggesting the coupling of Na(+) influx and Mg(2+) efflux. Na(+) influxes were different for normal and imipramine- (a putative inhibitor of the Na(+)/Mg(2+) transporter) containing solutions. FCCP induced a rapid increase in [Na(+)](i) in the normal solution, while the increase was gradual in the imipramine-containing solution. The rate of decrease of [Mg(2+)](i) in the imipramine-containing solution was also slower than that in the normal solution. From these results, we show that the main buffering mechanism for excess Mg(2+) depends on the Na(+)/Mg(2+) transporter in PC12 cells. PMID- 12646208 TI - Protein kinase-dependent phosphorylation of the Menkes copper P-type ATPase. AB - The Menkes copper-translocating P-type ATPase (ATP7A; MNK) is a key regulator of copper homeostasis in humans. It has a dual role in supplying copper to essential cuproenzymes in the trans-Golgi network (TGN) and effluxing copper from the cell. These functions are achieved through copper-regulated trafficking of MNK between the TGN and the plasma membrane. However, the exact mechanism(s) which regulate the localisation and biochemical functions of MNK are still unknown. Here we investigated copper-dependent phosphorylation of MNK by a putative protein kinase(s). We found that in the presence of elevated copper there was a substantial increase in phosphorylation of the wild-type MNK in vivo. The majority of copper-dependent phosphorylation was on serine residues in two phosphopeptides. In contrast, there was no up-regulation of phosphorylation of a non-trafficking MNK mutant with mutated cytosolic copper-binding sites. Our findings suggest a potentially important role of kinase-dependent phosphorylation in the regulation of function of the MNK protein. PMID- 12646209 TI - Molecular cloning and RNA expression of a novel Drosophila calpain, Calpain C. AB - The calpains are Ca(2+)-activated cysteine proteases whose biochemical properties have been extensively characterized in vitro. Less is known, however, about the physiological role of calpains. In this respect, Drosophila melanogaster is a useful experimental organism to study calpain activity and regulation in vivo. The sequencing of the fly genome has been recently completed and a novel calpain homologue has been identified in the CG3692 gene product. We embarked on the cloning and characterization of this putative novel calpain. We demonstrate that the actual calpain is different from the predicted protein and we provide experimental evidence for the correction of the genomic annotation. This novel protein, Calpain C, must be catalytically inactive, having mutated active site residues but is otherwise structurally similar to the other known fly calpains. Moreover, we analysed Calpain C RNA expression during Drosophila development by RT-PCR and RNA in situ hybridization, which revealed strong expression in the salivary glands. PMID- 12646210 TI - The eta isoform of protein kinase C inhibits UV-induced activation of caspase-3 in normal human keratinocytes. AB - Protein kinase C (PKC) fulfills a central role in the decision of cell fate in keratinocytes. Both PKC delta and PKC eta induce growth inhibition and differentiation of normal human keratinocytes (NHK). Here we show that PKC delta and PKC eta play opposite roles in UVB-induced apoptosis in NHK. PKC delta enhanced UVB-induced caspase-3 activity, while overexpression of PKC eta reduced it. In keeping with these observations, the dominant negative mutant of PKC delta significantly inhibited the activation of caspase-3, whereas dominant negative PKC eta increased it in a dose (MOI)-dependent manner. Unlike PKC delta, cleavage and translocation to mitochondria of PKC eta were not observed, resulting in no detection of cytochorome c release. Furthermore, UV-induced activation of p38 MAP kinase, which suppressed the caspase-3 activity in NHK, was blocked by dominant negative PKC eta. These findings suggest that PKC eta negatively regulates UV induced apoptosis through its localization, resistance to cleavage, and the p38 MAPK pathway. PMID- 12646211 TI - Molecular cloning and expression of the hepatitis delta virus genotype IIb genome. AB - Analysis of hepatitis delta virus (HDV) genome sequences has revealed multiple genotypes with different geographical distributions and associated disease patterns. To date, replication-competent cDNA clones of HDV genotypes I, II, and III have been reported. HDV genotypes I, II, and IIb have been found in Taiwan. Although full-length sequences of genotype IIb have been published, its replication competence in cultured cells has yet to be reported. In order to examine this, we obtained a full-length cDNA clone, Taiwan-IIb-1, from a Taiwanese HDV genotype IIb isolate. Comparison of the complete nucleic acid sequence of Taiwan-IIb-1 with previously published genotype IIb isolates indicated that Taiwan-IIb-1 shares 98% identity with another Taiwanese isolate and 92% identity with a Japanese isolate. Transfection of Taiwan-IIb-1 into COS7 cells resulted in accumulation of the HDV genome and appearance of delta antigens, showing that cloned HDV genotype IIb can replicate in cultured cells. PMID- 12646212 TI - Molecular identification of nicotinic acid receptor. AB - Nicotinic acid and its derivative, Acipimox, have been widely used in the treatment of hyperlipidemia. Pharmacological studies have demonstrated that they exert the beneficial effect through the activation of a Gi-protein-coupled receptor on adipocyte, which has remained elusive to date. Here we show that a novel GPCR, designated HM74b because of its high similarity to HM74, is a receptor for nicotinic acid. HM74b mRNA is found in human, murine, and rat adipose tissues. Nicotinic acid and Acipimox inhibit forskolin-stimulated intracellular cAMP accumulation in human HM74b-expressing cells and activate GTP gamma S binding in a dose-dependent manner. [3H]Nicotinic acid specifically binds to HM74b-expressing membrane and its binding is replaced by Acipimox. This finding will open a new phase of research on the physiological role of nicotinic acid and will be a clue to develop novel antihyperlipidemic drugs. PMID- 12646213 TI - Characterization of Grp1p, a novel cis-Golgi matrix protein. AB - A high copy suppressor screen with sec34-2, a temperature-sensitive mutant defective in the late stages of ER to Golgi transport, has resulted in the identification of a novel gene called GRP1 (also called RUD3). GRP1 encodes a hydrophilic yeast protein related to the mammalian Golgi matrix protein golgin 160. A large portion of the protein is predicted to form a coiled-coil structure. Although GRP1 is not essential for growth, the loss of Grp1p results in a growth defect at high temperature. GRP1 genetically interacts with several genes involved in vesicle targeting/fusion stages of ER to Golgi transport. Despite these interactions, pulse chase analysis using Grp1p-depleted cells did not reveal a significant delay in the transit of the vacuolar protease carboxypeptidase Y. Grp1p-depleted cells efficiently secreted invertase which was underglycosylated, suggesting some disturbance of Golgi function. Grp1p-GFP predominantly colocalizes with the cis-Golgi marker Och1p. Despite lacking a signal peptide and a significant stretch of hydrophobic amino acids, Grp1p pellets with membranes. It is extracted with 1M NaCl or 0.1M Na(2)CO(3) (pH 11.0), but is surprisingly insoluble in 1% Triton X-100. Grp1p does not recycle to the ER when forward transport is blocked and a cis-Golgi marker (Och1p-HA), but not a trans-Golgi marker (Chs5p-HA), became dispersed in grp1 Delta cells after 1.5h incubation at 38.5 degrees C. Together, these data suggest that Grp1p is a novel matrix protein that is involved in the structural organization of the cis-Golgi. PMID- 12646214 TI - Expression of adrenomedullin and its receptor by chondrocyte phenotype cells. AB - For clarifying a process of de-differentiation in culturing chondrocytes, the present study was undertaken to investigate the secretion of adrenomedullin (AM) by chondrocyte phenotype cells and whether or not AM effects this proliferation in a cAMP-dependent fashion. Chondrocyte phenotype cells expressed AM and the AM receptor, and secreted high concentration of AM into the culture medium. When added to cultures, AM increased the intracellular cAMP level and decreased the number of these cells in a similar concentration-dependent fashion. Addition of forskolin and dibutyryl-cAMP caused a significant decrease in the number of these cells. Furthermore, the effect of AM was inhibited by a cAMP-dependent protein kinase A inhibitor (H89). The present findings indicate that AM has an autocrine/paracrine type of anti-proliferative effect on these cells mediated via a cAMP-dependent pathway and raise the possibility that AM plays a role in the local modulation of a process of de-differentiation by culturing chondrocyte phenotype cells. PMID- 12646215 TI - Functional characterization of beta-ketoacyl-ACP reductase (FabG) from Plasmodium falciparum. AB - The malaria parasite, Plasmodium falciparum, unlike its human host, utilizes type II fatty acid synthesis, in which steps of fatty acid biosynthesis are catalyzed by independent enzymes. Due to this difference, the enzymes of this pathway are a potential target of newer antimalarials. Here we report the functional characterization of Plasmodium FabG expressed in Escherichia coli. The purified recombinant FabG from P. falciparum is soluble and active. The K(m) of the enzyme for acetoacetyl-CoA was estimated to be 75 microM with a V(max) of 0.0054 micromol/min/ml and a k(cat) value of 0.014s(-1). NADPH exhibited negative cooperativity for its interaction with FabG. We have also modeled P. falciparum FabG using Brassica napus FabG as the template. This model provides a structural rationale for the specificity of FabG towards its cofactor, NADPH. PMID- 12646216 TI - U-box proteins as a new family of ubiquitin ligases. AB - Ubiquitin-protein ligases (E3s) determine the substrate specificity of ubiquitylation and, until recently, had been classified into two families, the HECT and RING-finger families. The U-box is a domain of approximately 70 amino acids that is present in proteins from yeast to humans. The prototype U-box protein, yeast Ufd2, was identified as a ubiquitin chain assembly factor (E4) that cooperates with a ubiquitin-activating enzyme (E1), a ubiquitin-conjugating enzyme (E2), and an E3 to catalyze the formation of a ubiquitin chain on artificial substrates. We recently showed that mammalian U-box proteins, in conjunction with an E1 and an E2, mediate polyubiquitylation in the absence of a HECT type or RING-finger type E3. U-box proteins have thus been defined as a third family of E3s. We here review recent progress in the characterization of U box proteins and of their role in the quality control system that underlies the cellular stress response to the intracellular accumulation of abnormal proteins. PMID- 12646217 TI - Molecular cloning, biochemical and structural analysis of elongation factor-1 alpha from Leishmania donovani: comparison with the mammalian homologue. AB - The Src-homology 2 domain containing protein tyrosine phosphatase-1 (SHP-1) is involved in the pathogenesis of infection with Leishmania. Recently, we identified elongation factor-1 alpha (EF-1 alpha) from Leishmania donovani as a SHP-1 binding and activating protein [J. Biol. Chem. 277 (2002) 50190]. To characterize this apparent Leishmania virulence factor further, the cDNA encoding L. donovani EF-1 alpha was cloned and sequenced. Whereas nearly complete sequence conservation was observed amongst EF-1 alpha proteins from trypanosomatids, the deduced amino acid sequence of EF-1 alpha of L. donovani when compared to mammalian EF-1 alpha sequences showed a number of significant changes. Protein structure modeling-based upon the known crystal structure of EF-1 alpha for Saccharomyces cerevisiae-identified a hairpin loop present in mammalian EF-1 alpha and absent from the Leishmania protein which corresponded to a 12 amino acid deletion. Consistent with these structural differences, the sub-cellular distributions of L. donovani EF-1 alpha and host EF-1 alpha were strikingly different. Interestingly, infection of macrophages with L. donovani caused redistribution of host as well as pathogen EF-1 alpha. Since EF-1 alpha is essential for survival, the distinct biochemical and structural properties of Leishmania EF-1 alpha may provide a novel target for drug development. PMID- 12646218 TI - Role of protein kinase C alpha for uptake of unopsonized prey and phagosomal maturation in macrophages. AB - Protein kinase C alpha (PKC alpha) participates in F-actin remodeling during phagocytosis and phagosomal maturation in macrophages. Leishmania donovani promastigotes, which inhibit phagosomal maturation, cause accumulation of periphagosomal F-actin instead of the disassembly observed around other prey [Cell. Microbiol. 7 (2001) 439]. This accumulation is induced by promastigote lipophosphoglycan (LPG), which has several effects on macrophages including inhibition of PKC alpha. To investigate a possible connection between PKC alpha and LPG's effects on actin dynamics, we utilized RAW264.7 macrophages overexpressing dominant-negative PCK alpha (DN PKC alpha). We found increased cortical F-actin and decreased phagocytic capacity, as well as defective periphagosomal F-actin breakdown and inhibited phagosomal maturation in the DN PKC alpha-overexpressing cells, effects similar to those seen in controls subjected to LPG-coated prey. The results indicate that PKC alpha is involved in F-actin turnover in macrophages and that PKC alpha-dependent breakdown of periphagosomal F-actin is required for phagosomal maturation, and endorse the hypothesis that intracellular survival of L. donovani involves inhibition of PKC alpha by LPG. PMID- 12646219 TI - Inhibition of Plasmodium falciparum proliferation in vitro by antisense oligodeoxynucleotides against malarial topoisomerase II. AB - The development of new effective antimalarial agents is urgently needed due to the ineffectiveness of current drug regimes on the most virulent human malaria parasite Plasmodium falciparum. Antisense (AS) oligodeoxynucleotides (ODNs) have shown promise as chemotherapeutic agents. Phosphorothioate AS ODNs against different regions of P. falciparum topoisomerase II gene were investigated. Chloroquine- and pyrimethamine-resistant P. falciparum K1 strain was exposed to phosphorothioate AS ODNs for 48 h and growth was determined by flow cytometric assay or by microscopic assay. Exogenous delivery of phosphorothioate AS ODNs between 0.01 and 0.5 microM significantly inhibited parasite growth compared with sense sequence controls suggesting sequence specific inhibition. This inhibition was shown to occur during maturation stages, with optimal inhibition being detected after 36 h. These results should prove useful in future designs of novel antimalarial agents. PMID- 12646220 TI - Nitrosative stress on yeast: inhibition of glyoxalase-I and glyceraldehyde-3 phosphate dehydrogenase in the presence of GSNO. AB - Under nitrosative stressed condition intracellular GSNO accumulation is common to all cell types. Conserved NADH-dependent GSNO reductase was reported previously as an important cellular protective measure against this. In spite of the constitutive nature of the enzyme, we observed in vivo inactivation of two important enzymes-glyoxalase-I and glyceraldehyde-3-phosphate dehydrogenase under 5 mM GSNO stress in two budding yeasts, though with difference in their sensitivity. Former was more susceptible to inactivation in in vitro condition, too. In this study, we explored the competitive nature of yeast glyoxalase-I inhibition by GSNO. GSNO actually competes with GSH substrate-binding site of the enzyme. PMID- 12646221 TI - Inhibition of ubiquitin ligase Siah-1A by disabled-1. AB - Disabled-1 (Dab1) is a cytosolic adaptor protein that plays critical roles in cortical development. However, a detailed mechanism of action has not yet been clearly defined. Through yeast two-hybrid screening, we observed that mouse Siah 1A, an E3 ubiquitin ligase containing a RING finger motif, interacts with Dab1. Co-immunoprecipitation experiments and in vitro binding experiments both indicated direct interaction between Siah and Dab1. Steady-state expression of Siah was enhanced by the presence of Dab1 or lactacystin, a representative proteasomal inhibitor. Auto-ubiquitination of Siah was inhibited by the presence of Dab1, suggesting inhibition of Siah activity and subsequent increase of Siah expression by Dab1. Both Dab1-induced increase of steady-state expression of Deleted in colorectal cancer (DCC), one of the well-known substrates of Siah, and its inhibition by Siah Delta R suggest that Dab1 increases expression of DCC through inhibiting the activity of endogenous Siah. Our results suggest that Dab1 inhibits the activity of Siah. PMID- 12646223 TI - Norepinephrine and neuropeptide Y promote proliferation and collagen gene expression of hepatic myofibroblastic stellate cells. AB - The mechanisms initiating and perpetuating the fibrogenic response in the injured liver are not well understood. Hepatic stellate cells are activated by liver injury to become proliferative and fibrogenic myofibroblasts. Emerging evidence suggests that the sympathetic nervous system may play a role in the development of cirrhosis. It is not known, however, whether this requires a direct interaction between sympathetic neurotransmitters and stellate cell receptors, or results indirectly, from sympathetic effects on the vasculature. Using cultured hepatic stellate cells, we show that the sympathetic neurotransmitters, norepinephrine and neuropeptide Y, markedly stimulate the proliferation of activated, myofibroblastic, hepatic stellate cells. Norepinephrine, but not neuropeptide Y, also induces collagen gene expression. In conclusion, physiologically relevant concentrations of sympathetic neurotransmitters directly modulate the phenotype of hepatic stellate cells. This suggests that targeted interruption of sympathetic nervous system signaling in hepatic stellate cells may be useful in constraining the fibrogenic response to liver injury. PMID- 12646222 TI - Antioxidant activity of different dihydropyridines. AB - Lacidipine, a dihydropyridine-based calcium antagonist (DHP), has already been demonstrated to possess antioxidant activity and to reduce the intracellular production of reactive oxygen species (ROS). To verify if this effect is a peculiarity of this molecule, or belongs to other DHPs, the activity of lacidipine was compared with those of amlodipine, lercanidipine, nimodipine, and nifedipine. The DHPs were incorporated in bovine aortic endothelial cells (BAECs). Cu(2+)-oxidized LDL (ox-LDL, 5 microM) was incubated with BAECs for 5 min. 2',7'-Dichlorofluorescein (DCF) as expression of intracellular ROS production was measured by flow cytometry. Ox-LDL induced a strong increase in intracellular ROS formation (p<0.001) that was significantly reduced only with lacidipine and lercanidipine (p from <0.05 to <0.01); the effect of lacidipine, however, resulted in being much more evident than lercanidipine (p<0.01); amlodipine, nimodopine, and nifedipine had no effect on ROS formation. The lowest IC50s, i.e. the concentrations determining the 50% reduction of ROS, were obtained with lacidipine (p<0.01). The inhibitory effect of lacidipine on ox-LDL induced ROS production in endothelial cells is a peculiarity of this molecule through its antioxidant activity. PMID- 12646224 TI - The neurotoxic phospholipase A2 associates, through a non-phosphorylated binding motif, with 14-3-3 protein gamma and epsilon isoforms. AB - Two novel acceptors for ammodytoxin C, a presynaptically neurotoxic phospholipase A(2) from snake venom, have been purified from porcine cerebral cortex by a toxin affinity-based procedure. Using tandem mass spectrometry, the isolated acceptors were identified as 14-3-3 gamma and epsilon isoforms, highly conserved cytoplasmic proteins involved in the regulation of numerous physiological processes. The interaction between ammodytoxin C and 14-3-3 proteins is direct and not mediated by calmodulin, a high-affinity acceptor for both ammodytoxin C and 14-3-3 proteins, as demonstrated in pull-down experiments and by surface plasmon resonance. The latter technique gave an apparent dissociation constant of 1.0+/-0.2 microM for the interaction between chip-immobilized 14-3-3 and ammodytoxin C. 14-3-3 usually interacts with proteins through specific phospho Ser/Thr motifs. Ammodytoxin C is not a phospho-protein, therefore the interaction must occur through a non-phosphorylated binding site, most probably the KEESEK sequence at its C-terminal end. The interaction we describe suggests an explanation for the pathophysiological effects evoked by some secreted phospholipases A(2), such as the inhibition of protein phosphorylation, of terminal ion currents, and of neurotransmission, as well as the initiation of neuronal cell death, all processes regulated by 14-3-3 proteins. PMID- 12646225 TI - ZccR--a MerR-like regulator from Bordetella pertussis which responds to zinc, cadmium, and cobalt. AB - A transcriptional regulator of the MerR family encoded by Bordetella pertussis was characterized in Escherichia coli and in vitro. Uniquely, the regulator responded specifically to Zn(II), Cd(II), and Co(II) and was named ZccR. Gel shift assays confirmed that ZccR binds to an adjacent divergent promoter possessing an elongated spacer region of 19bp between the -10 and -35 elements, and that Zn(II), Co(II), and Cd(II) reduced the protein affinity for DNA. Site directed mutagenesis of four cysteine and six histidine residues of ZccR showed that the cysteine residues at positions 77, 112, and 122, conserved in many of the metal-responsive MerR-like regulators, were essential for induction. Mutagenesis of the histidine residues (positions 73, 87, 90, 126, 140, and 142) revealed that histidine residues at 90, 140, and 142 were required for full induction by all three metals. PMID- 12646226 TI - Altered antigenic disposition of peroxisomal urate oxidase in PEX5-defective Chinese hamster ovary cells. AB - Since Chinese hamster ovary (CHO) cells never express urate oxidase (UO), we tried to establish cell lines stably producing UO in order to elucidate the peroxisomal import process. The enzyme is a peroxisome targeting signal 1 (PTS1) protein harboring SKL motif at the carboxy-terminus [Biochem. Biophys. Res. Commun. 158 (1989) 991] and PEX5 protein (Pex5p) is supposed to be involved in the import process [Nat. Genet. 9 (1995) 115; J. Cell Biol. 130 (1995) 51]. We transfected a cDNA encoding rat UO into both wild type and PEX5-defective CHO cells to isolate each cell line stably producing the enzyme. While we examined the import process of UO in mutant cells, we noticed an interesting observation by using polyclonal antibody U1 or U2, which separately recognizes epitopes of UO. U1 antibody mainly interacts with epitopes in the amino-terminal region of UO. On the other hand, U2 antibody reacts with many epitopes distributed in the broad region of UO molecule. When UO produced in cultured cells was stained with U2 antibody, the enzyme was detected in peroxisomes of both wild type and PEX5 mutant cells. Whereas, U1 antibody stained the peroxisomal UO in wild type cells, but not in PEX5-mutant cells. These immunocytochemical observations suggest that the epitopes at the amino-terminal region of UO will be concealed in mutant cells. When the mutant cells were transfected with wild type PEX5 cDNA, U1 antibody came to react with UO in peroxisomes of mutant cells. The restoration indicates that the exposure of N-terminal epitopes of UO will depend upon the functional Pex5p. Immunoelectron microscopic observation showed that the peroxisomal import of UO was partially retarded in PEX5 mutant cells. The observation also supported the fact that UO was mainly localized in the peroxisomal matrix of wild type cells but in the membrane of mutant cells. PMID- 12646227 TI - Enhanced stability of alpha B-crystallin in the presence of small heat shock protein Hsp27. AB - Lens alpha-crystallin, alpha A- and alpha B-crystallin, and Hsp27 are members of the small heat shock protein family. Both alpha A- and alpha B-crystallin are expressed in the lens and serve as structural proteins and as chaperones, but alpha B-crystallin is also expressed in nonlenticular organs where Hsp27, rather than alpha A-crystallin, is expressed along with alpha B-crystallin. It is not known what additional function Hsp27 has besides as a heat shock protein, but it may serve, as alpha A-crystallin does in the lens, to stabilize alpha B crystallin. In this study, we investigate aspects on conformation and thermal stability for the mixture of Hsp27 and alpha B-crystallin. Size exclusion chromatography, circular dichroism (CD), and light scattering measurements indicated that Hsp27 prevented alpha B-crystallin from heat-induced structural changes and high molecular weight (HMW) aggregation. The results indicate that Hsp27 indeed promotes stability of alpha B-crystallin. PMID- 12646228 TI - Serotype and VP1 gene sequence of a foot-and-mouth disease virus from Hong Kong (2002). AB - The nucleotide sequence of the VP1 coding region of foot-and-mouth disease virus (FMDV) strain HKN/2002, isolated from a disease outbreak occurring in Hong Kong in February 2002, was determined and compared with the sequences of other FMDVs. The VP1 coding region was 639 nucleotides in length and encoded a protein of 213 amino acid residues. Comparison of the VP1 nucleotide sequence with those of other isolates indicated that HKN/2002 belonged to serotype O. A VP1-based sequence similarity tree of several South-east Asian FMDV-O isolates showed that HKN/2002 was most closely related to FMDV isolates found in Hong Kong from 1991 to 1999 and Taiwan in 1997. Comparison of the amino acid sequence of the major immunogenic region of HKN/2002 with that of the serotype O vaccine strain, O1/Manisa/Turkey/69, reveals significant similarity, indicating that current serotype O vaccines may offer some degree of protection against HKN/2002. PMID- 12646229 TI - HrcA is a negative regulator of the dnaK and groESL operons of Streptococcus pyogenes. AB - The genome of Streptococcus pyogenes, an important human pathogen, encodes homologs of the principal bacterial heat shock proteins DnaK and GroES, -EL, as well as HrcA, a negative regulator of dnaK and groESL expression in other Gram positive bacteria. Using nuclease protection assays to measure dnaK/groESL mRNA abundance and a "non-polar" insertion to disrupt hrcA, we demonstrate that heat shock triggers a 4- to 8-fold increase in dnaK and groESL-specific mRNAs within 5 min of the temperature shift and that HrcA is a negative regulator of S. pyogenes dnaK/groESL mRNA abundance in unstressed S. pyogenes. Although the loss of HrcA elevated dnaK and groESL mRNA levels under non-heat shock conditions, the relative abundance of these RNAs increased further in heat shocked S. pyogenes, suggesting an additional element contributing to their synthesis or stability. PMID- 12646230 TI - Multiple replication origins of the archaeon Halobacterium species NRC-1. AB - The genomic sequence of the halophilic archaeon Halobacterium NRC-1 has been analyzed by the Z curve method. The Z curve is a three-dimensional curve that uniquely represents a given DNA sequence. Based on the known behaviors of the Z curves for the archaea whose replication origins have been identified, the analysis of the Z curve for the genome of Halobacterium NRC-1 strongly suggests that the large genome has two replication origins, oriC1 (921,863-922,014) and oriC2 (1,806,444-1,807,229), which are located at two sharp peaks of the Z curve. These two regions are next to the cdc6 genes and contain multiple copies of stretches of G and C, i.e., ggggtgggg and ccccacccc, which may also be regarded as direct and inverted repeats. Based on the above analysis, a model of replication of Halobacterium NRC-1 with two replication origins and two termini has been proposed. The experimental confirmation of this model would constitute the first example of multiple replication origins of archaea, which will finally provide much insight into the understanding of replication mechanisms of eukaryotic organisms, including human. In addition, the potential multiple replication origins of the archaeon Sulfolobus solfataricus are suggested by the analysis based on the Z curve method. PMID- 12646231 TI - Mortalin-MPD (mevalonate pyrophosphate decarboxylase) interactions and their role in control of cellular proliferation. AB - Mortalin (mot-2/GRP75/PBP74/mthsp70) is a member of the hsp70 family of proteins and is differentially distributed in normal and immortal cells. It was shown to be involved in pathways to cell senescence and immortalization. To elucidate its functional aspects, a yeast interactive screen for mortalin (mot-2) binding proteins was performed. Mevalonate pyrophosphate decarboxylase (MPD) was identified as one of the mortalin binding partners. The interactions were confirmed in mammalian cells by two-hybrid assay and in vivo coimmunoprecipitation. MPD is known to furnish prenyl groups required for prenylation, protein modification that is essential for the activity of many proteins including p21(Ras) (Ras). We have examined the effect of MPD-mot-2 interactions on the level and activity of p21(Ras) and its downstream effectors, p44 and p42 MAP kinases (ERK1/ERK2), in Ras-Raf pathway. An overexpression of mot 2 resulted in reduced level of Ras and phosphorylated ERK2. These were rescued by co-expression of MPD from an exogenous promoter demonstrating a functional link between mot-2, MPD, and Ras. Ras and its oncogenic forms act as key players in controlling proliferation of normal and cancerous cells. Assigning mot-2 upstream of p21(Ras) offers an important mechanism for influence over cell proliferation. PMID- 12646232 TI - Improvement in blood lipid levels by dietary sn-1,3-diacylglycerol in young women with variants of lipid transporters 54T-FABP2 and -493g-MTP. AB - In a double-blind parallel-group study, serum lipids and visceral fat/total fat ratio in young women (n=49) with variants of lipid transporters, i.e., fatty acid binding protein 2 (FABP2) and microsomal triglyceride transfer protein (MTP), were analyzed by substituting dietary triacylglycerol (TAG) with sn-1,3 diacylglycerol (DAG). All subjects, including some with the hyperlipidemia-prone genotypes Ala54Thr of FABP2 and c-493g of MTP, received DAG or TAG (20 g/day) for 8 weeks. Reductions of serum lipids from weeks 4 to 8 in FABP2-Ala54Thr heterozygotes and MTP -493g homozygotes were significantly different between the DAG and TAG groups (p<0.05, p<0.01). Visceral fat/total fat (%), as determined by computed tomography (CT), was lower in FABP2-Ala54Thr heterozygotes (p<0.05) of the DAG group. The apoCII/CIII ratio was higher in the DAG group than in the TAG group (p<0.01). Other variants of lipid metabolism, including peroxisome proliferator activated receptors (PPARs) alpha and gamma and SREBP cleavage activating protein (SCAP), were only slightly affected by dietary DAG. CONCLUSION: improvement of serum lipid profiles and visceral fat/total fat ratio (CT) was potentiated by DAG intake in subjects with hyperlipidemia-prone genotypes (Ala54Thr heterozygotes of FABP2 and -493g homozygotes of MTP). PMID- 12646233 TI - Large-scale search of SNPs for type 2 DM susceptibility genes in a Japanese population. AB - The etiology of type 2 diabetes (DM) is polygenic. We investigated here genes and polymorphisms that associate with DM in the Japanese population. Single nucleotide polymorphisms (SNPs) of 398 derived from 120 candidate genes were examined for association with DM in a population-based case-control study. The study group consisted of 148 cases and 227 controls recruited from Funagata, Japan. No evident subpopulation structure was detected for the tested population. The association tests were conducted with standard allele positivity tables (chi(2) tests) between SNP genotype frequency and case-control status. The independent association of the SNPs from serum triglyceride levels and body mass index was examined by multiple logistic regression analysis. A value of P<0.01 was accepted as statistically significant. Six genes (met proto-oncogene, ATP binding cassette transporter A1, fatty acid binding protein 2, LDL receptor defect C complementing, aldolase B, and sulfonylurea receptor) were shown to be associated with DM. PMID- 12646234 TI - Methyl-CpG-DNA binding proteins in human prostate cancer: expression of CXXC sequence containing MBD1 and repression of MBD2 and MeCP2. AB - We analyzed gene expression of MBD1, MBD2, MBD3, MBD4, and MeCP2 and protein expression of MBD1, MBD2, and MeCP2 in prostate cancer cell lines, benign prostate epithelium (BPH-1) cell line, 49 BPH tissues, and 46 prostate cancer tissues. The results of this study demonstrate that MBD2 gene is expressed in all samples and MeCP2 gene is expressed in all cancer cell lines but not in BPH-1 cell line. However, there was no protein expression for MBD2 and MeCP2 in cancer cell lines and cancer tissues. For CXXC sequence containing MBD1, both protein and mRNA were expressed in cancer cell lines, cancer tissues, BPH-1 cell line, and BPH tissues. We observed that, in BPH tissues and low-grade cancer tissues, MBD1 protein expression was very high and gradually decreased with increase of cancer grade. Treatment of cancer cell lines with proteasome inhibitor (MG-132) did not restore expression of MBD2 and MeCP2 proteins. When prostate cancer cell lines were treated with hypomethylating agent, 5-aza-2(')-deoxycytidine (DNMT inhibitor), HDAC1 and HDAC2 expression was decreased. This is the first report demonstrating that CXXC sequence containing MBD1 is overexpressed and can be the major factor of hypermethylated chromatin segments through HDAC1/2 translocation and histone deacetylation in human prostate cancer. PMID- 12646236 TI - Mouse mast cell protease-1 cleaves angiotensin I to form angiotensin II. AB - The ability to convert angiotensin (Ang) I to Ang II was compared between human alpha-chymase and two mouse beta-chymases, mouse mast cell protease (mMCP)-1 and mMCP-4. Human chymase hydrolyzed Ang I to produce Ang II without further degradation. mMCP-1 similarly generated Ang II from Ang I in a time-dependent manner and the formation of the fragment other than Ang II was marginal. In contrast, mMCP-4 hydrolyzed Ang I at two sites, Tyr(4)-Ile(5) and Phe(8)-His(9), with Ang II formation being tentative. Consistently, mMCP-4 but not human chymase hydrolyzed Ang II and mMCP-1 showed little hydrolytic activity against Ang II. These data suggest that not only human chymase but also mMCP-1 might possess a physiological role in Ang II formation. Our findings also imply that the Ang converting activity of chymase may not be related to the categorization of chymase into alpha- or beta-type based on their primary structure. PMID- 12646235 TI - Molecular cloning of mammalian Spred-3 which suppresses tyrosine kinase-mediated Erk activation. AB - We have reported on Spred-1 and Spred-2, which inhibit MAP kinase activation by interacting with c-kit and ras/raf. Here, we report the cloning of a third member in this family, Spred-3. Spred-3 is expressed exclusively in the brain and its gene locates in chromosome 19q13.13 in human. Like Spred-1 and -2, Spred-3 contains an EVH1 domain in the N-terminus and a Sprouty-related cysteine-rich region (SPR domain) in the C-terminus that is necessary for membrane localization. However, Spred-3 does not possess a functional c-kit binding domain (KBD), since the critical amino acid Arg residue in this region was replaced with Gly in Spred-3. Although Spred-3 suppressed growth factor-induced MAP kinase (Erk) activation, inhibitory activity of Spred-3 was lower than that of Spred-1 or Spred-2. By the analysis of chimeric molecules between Spred-3 and Spred-1, we found that the SPR domain, rather than KBD, is responsible for efficient Erk suppression. The finding of Spred-3 revealed the presence of a novel family of regulators for the Ras/MAP kinase pathway, each member of which may have different specificities for extracellular signals. PMID- 12646237 TI - Altered cAMP signaling induced by lysophosphatidic acid in senescent human diploid fibroblasts. AB - Lysophosphatidic acid (LPA) is a lipid mitogen that acts through G-protein coupled receptors. LPA responsiveness has been reported to be dependent on the senescent state of the cells. To solve the mechanism underlying, we observed LPA dependent cAMP status and found its age-dependent contrasting profile such as high level of cAMP in the senescent cells vs its low level in the young cells. In order to clarify the molecular mechanism of the ageing effect, we examined various molecular species involved in the cAMP signaling pathway by semi quantitative RT-PCR. EDG-1 and EDG-4 were unchanged, but EDG-2 and EDG-7 were reduced with age. Senescent cells showed a partial reduction of Gi1, Gi2, and Gi3, but no change in the level of Gq. Decreased Gis and Gi-coupled LPA receptors may reduce the inhibitory effect of Gi alpha on adenylyl cyclases (ACs), resulting in cAMP accumulation via activation of adenylyl cyclase in senescent fibroblasts. We also observed an age-dependent increase in some of AC isoforms: II, IV, and VI. In conclusion, multiple changes in the cAMP signaling pathway of the senescent cells might explain the altered responsiveness to the mitogenic stimuli. PMID- 12646238 TI - Vasopressin-SV40 T antigen expression in transgenic mice induces brain tumor and lymphoma. AB - In order to understand the importance of various cis-acting elements in regulating VP gene expression, transgenic mice regulated by VP constructs were produced containing 3.8 kb of the 5' flanking region and all the exons and introns in the mouse VP gene, which was fused at the end of exon 3 to an SV40 T antigen (Tag). In the transgenic mice by the pVPSV.IGR3.6 construct, all the six transgenic mice died at the age of 2-6 weeks. In the transgenic mice by pVPSV.IGR2.1, 21% of them had brain tumors at 5 weeks and 100% of the mice had brain tumors after 24 weeks. Histological analysis of the transgenic mice revealed primitive neuroectodermal tumors (PNET) in the brain and lymphoma in the spleen and lymph nodes. The phenotype differences between the two transgenic mice suggest that tissue-specific expression might be regulated by cis-acting elements in the 1.5-kb of the 3(') flanking region, which are not contained in pVPSV.IGR2.1. In conclusion, pVPSV.IGR2.1 mice will be a valuable mouse model system for investigating PNET tumorigenesis in the brain and lymphoma in the lymph nodes and spleen. PMID- 12646240 TI - Uncoupling of protein kinase D from suppression of EGF-dependent c-Jun phosphorylation in cancer cells. AB - Protein kinase D (PKD) has been established as a negative modulator of the c-Jun N-terminal kinase (JNK) signaling pathway. We previously demonstrated that induced expression of constitutively active PKD (PKD-S744/748E) that mimics phosphorylation by PKC is sufficient to attenuate epidermal growth factor (EGF) stimulated c-Jun Ser 63 phosphorylation, a natural substrate of JNK, in HEK 293 cells. Because the JNK pathway has been implicated in sustaining both lung and pancreatic cancerous phenotypes, we have utilized stable inducible expression of PKD-S744/748E in clones of A549 non-small cell lung cancer (NSCLC) and Panc1, pancreatic cancer cells to determine its effects on JNK signaling in the context of the cancerous phenotype. In contrast to HEK 293 cells, induced expression of PKD-S744/748E in either A549 NSCLC or Panc1 cells failed to attenuate EGF dependent phosphorylation of c-Jun, indicating that EGF stimulated JNK phosphorylation of c-Jun is uncoupled from PKD suppression in these cancer cells. PMID- 12646239 TI - A peptide encoded by exon 6 of VEGF (EG3306) inhibits VEGF-induced angiogenesis in vitro and ischaemic retinal neovascularisation in vivo. AB - VEGF is an important mediator of pathological angiogenesis in the eye and is a target for the development of novel anti-angiogenic molecules. In a previous study we identified 12-amino acid peptides derived from exon 6 of VEGF that inhibited VEGF binding to its receptors in HUVECs, endothelial cell functions, and in vitro angiogenesis. Screening of a series of truncated peptides corresponding to the inhibitory region of exon 6 identified a seven amino acid residue peptide, RKRKKSR, as the minimum exon 6-encoded sequence which retains the ability to inhibit VEGF receptor binding and angiogenesis in vitro. The effect of the seven-residue peptide was examined in a mouse model of ischaemic retinal neovascularisation. Administration of the peptide caused a 50% inhibition of retinal neovascularisation and was as effective in inhibiting ischaemic angiogenesis as soluble Flt-1 adenovirus. These results demonstrate that a seven amino acid VEGF exon 6-derived peptide is an effective inhibitor of ocular neovascularisation in vivo, and may have applications in the treatment of pathophysiological ocular neovascularisation in human disease. PMID- 12646242 TI - The regulatory effect of hydrogen sulfide on hypoxic pulmonary hypertension in rats. AB - Hypoxic pulmonary hypertension (HPH) is an important pathophysiological process. The mechanism of HPH is still not fully understood. Recent studies showed that hydrogen sulfide (H(2)S) could relax vascular smooth muscles and inhibit the proliferation of cultured vascular smooth muscle cells. Our study showed that both the gene expression of cystathionine gamma-lyase (CSE), one of the H(2)S generating enzymes, and the activity of CSE were suppressed in lung tissues during HPH. And the plasma level of H(2)S was decreased during HPH. Exogenous supply of H(2)S could increase the plasma level of H(2)S, enhance CSE activity, and up-regulate CSE gene expression in lung tissue. At the same time, exogenous supply of H(2)S could oppose the elevation of pulmonary arterial pressure and lessen the pulmonary vascular structure remodeling during HPH. The results showed that endogenous H(2)S system was involved and exogenous H(2)S could exert beneficial effect on the pathogenesis of HPH. PMID- 12646241 TI - Vanadate enhances leptin-induced activation of JAK/STAT pathway in CHO cells. AB - Leptin, the product of the ob gene, is an adipocyte-derived hormone that plays a key role in the control of food intake and energy expenditure. Leptin acts through receptors that belong to a member of the class I cytokine receptor family. It has been demonstrated that the SH2 domain-containing tyrosine phosphatase 2 (SHP-2) negatively regulates STAT3-mediated transcriptional activation through long form leptin receptor (OBRb). Vanadate has been shown to be a potent and selective inhibitor of PTPase activity in vitro. In this study, we have demonstrated that vanadate increases leptin-induced JAK2 and STAT3 phosphorylation in CHO cells expressing OBRb. The increased leptin-dependent luciferase activity of SOCS3 gene was also seen in vanadate-treated cell. Furthermore, vanadate reversed the inhibitory effects of SOCS3 on leptin-induced STAT3 phosphorylation. The present findings suggest that PTP inhibitors including vanadate and vanadate-derived compounds could be used as a therapeutic agent in the treatment of obesity. PMID- 12646243 TI - Intracellular redistribution of protein kinase D2 in response to G-protein coupled receptor agonists. AB - The protein kinase D (PKD) family consists of three serine/threonine protein kinases: PKC mu/PKD, PKD2, and PKC nu/PKD3. While PKD has been the focus of most studies to date, no information is available on the intracellular distribution of PKD2. Consequently, we examined the mechanism that regulates its intracellular distribution in human pancreatic carcinoma Panc-1 cells. Analysis of the intracellular steady-state distribution of fluorescent-tagged PKD2 in unstimulated cells indicated that this kinase is predominantly cytoplasmic. Cell stimulation with the G protein-coupled receptor agonist neurotensin induced a rapid and reversible plasma membrane translocation of PKD2 by a mechanism that requires PKC activity. In contrast to the other PKD isoenzymes, PKD2 activation did not induce its redistribution from the cytoplasm to the nucleus. Thus, this study demonstrates that the regulation of the distribution of PKD2 is distinct from other PKD isoenzymes, and suggests that the differential spatio-temporal localization of these signaling molecules regulates their specific signaling properties. PMID- 12646244 TI - The appearance of truncated cyclin A2 correlates with differentiation of mouse embryonic stem cells. AB - The presence of a form of cyclin A2 with an N-terminal truncation has recently been reported in various murine cell lines and tissues. The truncated cyclin A2 binds to and activates the cyclin-dependent kinase 2 (CDK2). However, CDK2 bound by the truncated cyclin A2 is located in the cytoplasm in contrast to CDK2 bound to full-length cyclin A2, which is in the nucleus. Here, we show that proliferating mouse embryonic stem cells (ES cells) contain very little truncated cyclin A2 but as the cells are induced to differentiate the amount of truncated cyclin A2 increases. The expression pattern of truncated cyclin A2 was the same in p27(Kip1) -/- differentiating ES cells as in the differentiating wild-type cells. We conclude that p27(Kip1) is not necessary for the proteolytic cleavage that gives rise to the truncated form of cyclin A2 in differentiating ES cells and that this post-translational modification is not a function of the cell density but is correlated with differentiation. PMID- 12646245 TI - Overexpression of the CT GalNAc transferase inhibits muscular dystrophy in a cleavage-resistant dystroglycan mutant mouse. AB - Transgenic mice that express dystroglycan containing a serine to alanine point mutation at the normal site of cleavage (DG(S654A)) in their skeletal muscles fail to express endogenously cleaved dystroglycan and have muscular dystrophy [Neuromusc. Disord., in press]. Dystrophic DG(S654A) muscles have reduced binding of antibodies, including VIA4-1, that recognize carbohydrate antigens on alpha dystroglycan, a finding similar to muscles in some forms of congenital muscular dystrophy. Here we describe one DG(S654A) transgenic line where VIA4-1 antibody binding is absent in skeletal muscle. In theory, the absence of this carbohydrate antigen should inhibit later glycosylation events that would occur on the structure or structures this antibody binds to. One such modification is likely to be the CT carbohydrate antigen, which is present on alpha dystroglycan in muscles overexpressing the CT GalNAc transferase [Dev. Biol. 242 (2002) 58]. To test the relationship between the VIA4-1 and CT carbohydrate antigens, we made DG(S654A)/CT GalNAc transferase (DG(S654A)/CT) transgenic mice. Surprisingly, dystroglycan was cleaved, and alpha dystroglycan was glycosylated with the VIA4-1 antigen, in DG(S654A)/CT muscles. In addition, muscles in DG(S654A)/CT transgenic mice had little or no evidence of muscular dystrophy when compared to DG(S654A) littermates. These experiments demonstrate that the CT GalNAc transferase can affect the post-translational processing of dystroglycan and the extent of muscular dystrophy even in muscles where the VIA4-1 antigen is not present. PMID- 12646246 TI - Interleukin-6 gene expression is increased in insulin-resistant rat skeletal muscle following insulin stimulation. AB - IL-6 expression in skeletal muscle is stimulated by contractions. We sought to examine whether hyperinsulinaemia increases IL-6 mRNA in skeletal muscle and whether any increase is modified in insulin resistant muscle. We hypothesized that intramuscular IL-6 mRNA would be increased in response to insulin, but such an affect would be unaffected by insulin resistance because the primary insulin sensitive signalling protein responsible for activating IL-6 functions normally in insulin resistant muscle. Transgenic rats over-expressing the gluconeogenic regulatory enzyme phosphoenolpyruvate carboxykinase (PEPCK) were studied. White gastrocnemius muscle samples were obtained under hyperinsulinaemic, euglycaemic clamp (4 mU kg(-1)min(-1) insulin, plasma glucose concentration 4-6 mmol L(-1)) and basal conditions in both PEPCK (basal n=4; insulin n=5) and wild-type (CON) (basal n=5; insulin n=4) rats, which were previously injected with a bolus of 2 [1-14C]deoxyglucose (2-DG) into the carotid artery. Muscle samples were assayed for 2-DG uptake and IL-6 mRNA. No differences in 2-DG uptake or IL-6 mRNA were observed when comparing groups under basal conditions. Under clamp conditions, 2 DG uptake was lower (P<0.05) in PEPCK compared with CON. Insulin stimulation in CON did not change IL-6 mRNA compared with basal levels. In contrast, there was an approximately 8-fold increase (P<0.05) in IL-6 mRNA in insulin-stimulated PEPCK compared with CON basal levels. Insulin stimulation increases IL-6 gene expression in insulin resistant, but not healthy, skeletal muscle, suggesting that IL-6 expression in skeletal muscle is sensitive to changes in insulin in circumstances of insulin resistance. It is likely that the differences observed when comparing healthy with insulin resistant muscle are due to the differential activation of insulin sensitive signalling proteins responsible for activating IL 6. PMID- 12646248 TI - The structure and function of human dipeptidyl peptidase IV, possessing a unique eight-bladed beta-propeller fold. AB - Dipeptidyl peptidase IV (DPPIV) is a serine protease, a member of the prolyl oligopeptidase (POP) family, and has been implicated in several diseases. Therefore, the development of DPPIV selective inhibitors, which are able to control the biological function of DPPIV, is important. We determined the crystal structure of human DPPIV at 2.6A resolution. The molecule consists of a unique eight-bladed beta-propeller domain in the N-terminal region and a serine protease domain in the C-terminal region. Also, the large "cave" structure, which is thought to control the access of the substrate, is found on the side of the beta propeller fold. Comparison of the overall amino acid sequence between human DPPIV and POP shows low homology (12.9%). In this paper, we report the structure of human DPPIV, especially focusing on a unique eight-bladed beta-propeller domain. We also discuss the way for the access of the substrate to this domain. PMID- 12646247 TI - CBP/p300 induction is required for retinoic acid sensitivity in human mammary cells. AB - The coactivators CBP and p300 are recruited by retinoic acid receptors (RARs) during retinoid mediated transcriptional regulation. To assess the role of CBP/p300 in all-trans-retinoic acid (ATRA)-mediated growth arrest in mammary epithelial cells, two systems were tested: (1) ATRA resistant MCF-7 cells were transduced with a functional RAR-beta 2; (2) normal human mammary epithelial cells (HMECs) were transduced with a pan-RAR dominant negative, RAR-alpha 403. Expression of RAR-beta 2 in MCF-7 cells resulted in increased sensitivity to ATRA induced growth arrest and correlated with induction of CBP/p300 mRNA and protein. Inhibition of RAR function in HMECs resulted in resistance to ATRA-induced growth arrest and loss of CBP/p300 induction. Antisense suppression of CBP/p300 in HMECs resulted in decreased retinoic acid response element reporter trans-activation and decreased ATRA-mediated growth arrest. Thus, in human mammary epithelial cells, CBP/p300 were both modulated by an ATRA signaling pathway and were required for a normal response to ATRA. PMID- 12646249 TI - Involvement of Rho-kinase in P2Y-receptor-mediated contraction of renal glomeruli. AB - The involvement of Rho-kinase in P2Y-receptor induced contraction of isolated rat renal glomeruli was investigated. The contraction effects have been investigated based on changes in the intracapillary volume of isolated glomeruli. ATP was found to induce time- and concentration-dependent contraction of isolated glomeruli. Other tested nucleotides (ADP, UTP) and ATP analogues (beta,gamma methylene-ATP, 2-methylothio-ATP) contracted glomeruli in similar magnitude whereas AMP had no effect. Furthermore, the contractive effect of ATP was prevented in the presence of an antagonist of P2Y-receptors, reactive blue 2. However, a selective antagonist of A1-receptors, 8-cyclopentyl-1,3 dipropylxanthine (DPCPX), had no effect. Contraction induced by ATP, ADP, and UTP, in contrast to 2-methylothio-ATP and beta,gamma-methylene-ATP, was prevented in the presence of Rho-kinase's inhibitor, (R)-(+)-trans-N-(4-pyridyl)-4-(1 aminoethyl)-cyclohexanecarboxamide dihydrochloride monohydrate (Y-27632). These findings suggest the involvement of Rho-kinase pathways in P2Y-induced contraction of isolated glomeruli. PMID- 12646251 TI - Cytochrome c decelerates channel kinetics of negatively charged gramicidin due to electrostatic interaction. AB - The effect of cytochrome c on the kinetic properties of ion channels formed by O pyromellitylgramicidin (OPg), the negatively charged analogue of gramicidin A (gA), in bilayer lipid membranes was studied by the method of sensitized photoinactivation. The addition of cytochrome c to both sides of the membrane caused substantial deceleration of the photoinactivation kinetics of OPg channels which expose three negative charges to the aqueous phase at both sides of the membrane. By contrast, the gA photoinactivation kinetics was unaltered by the addition of cytochrome c. Based on the sensitivity of the observed effect to the ionic strength of the bathing solution, the cytochrome c-induced deceleration of the OPg photoinactivation kinetics reflecting the increase in the OPg channel lifetime was ascribed to electrostatic interaction of positive charges of cytochrome c with negative charges of OPg that resulted in channel clustering. Formation of clusters of OPg channels was previously inferred to explain the polylysine effect on the OPg channel kinetics. The decelerating effect of cytochrome c on OPg channels was observed only at a high number of OPg channels in the membrane, thus suggesting that the interaction between cytochrome c and the charged transmembrane protein requires sufficiently high negative charge density on the surface of the membrane. PMID- 12646250 TI - Ergothioneine inhibits oxidative stress- and TNF-alpha-induced NF-kappa B activation and interleukin-8 release in alveolar epithelial cells. AB - Oxidants and inflammatory mediators such as tumour necrosis factor-alpha (TNF alpha) activate transcription factors such as NF-kappa B. Interleukin-8 (IL-8) is a ubiquitous inflammatory chemokine that mediates a multitude of inflammatory events in the lung. Ergothioneine is a naturally occurring thiol compound, which possesses antioxidant property. The aim of this study was to determine whether ergothioneine can inhibit the hydrogen peroxide (H(2)O(2))- and TNF-alpha mediated activation of NF-kappa B and the release of IL-8 in human alveolar epithelial cells (A549). Treatment of A549 cells with H(2)O(2) (100 microM) and TNF-alpha (10 ng/ml) significantly increased NF-kappa B activation using a reporter assay. Ergothioneine inhibited both H(2)O(2)- and TNF-alpha-mediated activation of NF-kappa B. Both H(2)O(2) and TNF-alpha significantly increased IL 8 release, which was inhibited by pre-treatment of A549 cells with ergothioneine compared to the control untreated cells. Ergothioneine also abolished the transcriptional activation of IL-8 in an IL-8-chloramphenicol acetyltransferase (CAT) reporter system, transfected into A549 cells. This indicates a molecular mechanism for the anti-inflammatory effects of ergothioneine. PMID- 12646252 TI - Mammalian Numb is a target protein of Mdm2, ubiquitin ligase. AB - Drosophila Numb protein functions as an antagonist against Notch signal. The expression of this protein is asymmetrical in divided cells and thought to be involved in the neural cell differentiation and/or cell fate. Human homologue of Numb (hNumb) was cloned as Mdm2-binding protein by yeast two-hybrid screening. Since Mdm2 is an oncoprotein and has ubiquitin ligase activity toward tumor suppressor p53, we assessed to find out whether Mdm2 ubiquitinylates the hNumb protein. The recombinant hNumb expressed in Sf-9 cells using baculovirus protein expression system bound to Mdm2 in vitro. When hNumb was subjected to in vitro ubiquitinylation assay system, which contains E1, E2, or UbcH5c, and Mdm2, hNumb was ubiquitinylated as efficiently as the p53 protein. However, when the Ring finger domain mutant of Mdm2 was used in place of wild-type Mdm2, hNumb was not ubiquitinylated. Furthermore, when U2OS cells were co-transfected with hNumb and Mdm2, the hNumb protein was ubiquitinylated and degraded. These data strongly suggest that Mdm2 functions as the ubiquitin ligase toward hNumb and that it induces its degradation in intact cells. PMID- 12646253 TI - Platelet aggregation by membrane-expressed A1 domains of von Willebrand Factor is dependent on residues Asp 560 and Gly 561. AB - We have previously demonstrated that the isolated von Willebrand Factor (vWF)-A1 domain can activate platelets in a GPIb-dependent manner. Here we evaluated the functional impact of targeted point mutations Gly561His (G1324(561)H), an analog of a previously described 2M von Willebrand disease variant, and Asp560Ser (D1323(560)S) in the model of the membrane expressed A1-domain. Platelet aggregation in response to COS-7-cells stably transfected with wild type A1 domain was abrogated by both substitutions. Ristocetin did not increase the aggregatory potential of mutant vWF-A1, in contrast to native forms. Botrocetin boosted the aggregatory responses of all A1-domains tested. These data suggest that G1324(561) and D1323(560) comprise part of the GPIb binding motif essential for subsequent platelet aggregation. Botrocetin seems to alter the potential of vWF for GPIb interaction independently of that motif. The experimental system tested here provides a rapid and reproducible approach for the functional analysis of isolated A1-domain interactions with platelet-GPIb. PMID- 12646254 TI - Involvement of tonB-exbBD1D2 operon in infection of Xanthomonas campestris phage phi L7. AB - phi L7 is a lytic bacteriophage infecting Xanthomonas campestris pv. campestris, a Gram-negative bacterium producing xanthan gum and causing black rot in crucifers. A mutant resistant to phi L7 was isolated by Tn5 mutagenesis. Sequence analysis indicated that the gene responsible for the mutation is tonB encoding an inner membrane protein previously shown to be required for iron uptake and pathogenesis. This gene is clustered with three other genes, tonB-exbB-exbD1 exbD2. Results of insertional mutations, DNA and protein sequence analyses, phage sensitivity tests, transfection tests, complementation tests, and phage adsorption assays together with the cellular location of the proteins indicate that TonB, ExbB, and ExbD1 are essential for penetration of phage phi L7. The genome organization, structural features of the tonB-exb region, and transcriptional analyses including Northern hybridization, reporter assays, and primer extension together indicate that the four genes are organized into an operon. PMID- 12646255 TI - Effect of DNA length and H4 acetylation on the thermal stability of reconstituted nucleosome particles. AB - To examine the factors involved with nucleosome stability, we reconstituted nonacetylated particles containing various lengths (192, 162, and 152 base pairs) of DNA onto the Lytechinus variegatus nucleosome positioning sequence in the absence of linker histone. We characterized the particles and examined their thermal stability. DNA of less than chromatosome length (168 base pairs) produces particles with altered denaturation profiles, possibly caused by histone rearrangement in those core-like particles. We also examined the effects of tetra acetylation of histone H4 on the thermal stability of reconstituted nucleosome particles. Tetra-acetylation of H4 reduces the nucleosome thermal stability by 0.8 degrees C as compared with nonacetylated particles. This difference is close to values published comparing bulk nonacetylated nucleosomes and core particles to ones enriched for core histone acetylation, suggesting that H4 acetylation has a dominant effect on nucleosome particle energetics. PMID- 12646256 TI - Overexpression of hepatocyte growth factor receptor in renal carcinoma cells indirectly stimulates tumor growth in vivo. AB - We examined the role of increased expression of HGFR kinase in in vivo growth of renal carcinoma. Human renal carcinoma cell line, ACHN cells, was transfected with plasmid encoding wild-type HGFR gene to generate cell lines with increased HGFR protein. ACHN cells with elevated HGFR expression, denoted clones 8 and 10, respectively, showed higher basal kinase activities of HGFR and PI3-kinase than those of empty-vector (mock)-transfected cells. Clone 8 and 10 cells grew similar to mock cells in culture. In mice, tumors of these clones grew more rapidly than those of mock cells. Microvessel density of clone 8 or 10 tumors was higher than that of mock tumors. Clone 8 and 10 cells secreted vascular endothelial growth factor-A (VEGF-A) more than mock cells and the secretion was PI3-kinase inhibitor, LY294002-sensitive. Anti-VEGF-A neutralizing antibody significantly inhibited tumor growth of clones 8 and 10 in mice. These results indicate for the first time that overexpression of HGFR tyrosine kinase in renal carcinoma cells participates in rapid tumor growth in vivo. PMID- 12646257 TI - Fatty acid synthase drives the synthesis of phospholipids partitioning into detergent-resistant membrane microdomains. AB - Fatty acid synthase (FAS) is a key metabolic enzyme catalyzing the synthesis of long-chain saturated fatty acids. It plays a central role in the production of surfactant in fetal lungs, in the supply of fatty components of milk, and in the conversion and storage of energy in liver and adipose tissue. Remarkably high levels of FAS expression are found in the majority of human epithelial cancers. As the role of FAS in cancer cells remains largely unknown, we have initiated studies to assess the fate of newly synthesized lipids in cancer cells and have estimated the contribution of FAS to the synthesis of specific lipid classes by treating the cells with small interfering RNAs targeting FAS. Here, we show that in cancer cells FAS plays a major role in the synthesis of phospholipids partitioning into detergent-resistant membrane microdomains. These are raft aggregates implicated in key cellular processes including signal transduction, intracellular trafficking, cell polarization, and cell migration. These findings reveal a novel role for FAS, provide important new insights into the otherwise poorly understood mechanisms underlying the control of lipid composition of membrane microdomains, and point to a link between FAS overexpression and dysregulation of membrane composition and functioning in tumor cells. PMID- 12646258 TI - ECRG2, a novel candidate of tumor suppressor gene in the esophageal carcinoma, interacts directly with metallothionein 2A and links to apoptosis. AB - Esophageal cancer related gene 2 (ECRG2) is a novel candidate of the tumor suppressor gene identified from human esophagus. To study the biological role of the ECRG2 gene, we performed a GAL4-based yeast two-hybrid screening of a human fetal liver cDNA library. Using the ECRG2 cDNA as bait, we identified nine putative clones as associated proteins. The interaction of ECRG2 and metallothionein 2A (MT2A) was confirmed by glutathione S-transferase pull-down assays in vitro and co-immunoprecipitation experiments in vivo. ECRG2 co localized with MT2A mostly to nuclei and slightly to cytoplasm, as shown by confocal microscopy. Transfection of ECRG2 gene inhibited cell proliferation and induced apoptosis in esophageal cancer cells. In the co-transfection of ECRG2 and MT2A assays, cell proliferation was inhibited and apoptosis was slightly induced compared with control groups. When we used antisense MT2A to interdict the effect of MT2A, the inhibition of cell proliferation and induction of apoptosis were significantly enhanced. When we used antisense ECRG2 to interdict the effect of ECRG2 in the group of Bel7402 cells co-transfected with ECRG2 and MT2A, the inhibition of cell proliferation and induction of apoptosis disappeared. The results provide evidence for ECRG2 in esophageal cancer cells acting as a bifunctional protein associated with the regulation of cell proliferation and induction of apoptosis. ECRG2 might reduce the function of MT2A on the regulation of cell proliferation and induction of apoptosis. The physical interaction of ECRG2 and MT2A may play an important role in the carcinogenesis of esophageal cancer. PMID- 12646260 TI - The dragging emergence of the P450 cytochrome. PMID- 12646261 TI - Glutathione, S-substituted glutathiones, and leukotriene C4 as substrates for peptidylglycine alpha-amidating monooxygenase. AB - The C-terminal alpha-amide moiety of most peptide hormones arises by the posttranslational cleavage of a glycine-extended precursor in a reaction catalyzed by bifunctional peptidylglycine alpha-amidating monooxygenase (PAM). Glutathione and the S-alkylated glutathiones have a C-terminal glycine and are, thus, potential substrates for PAM. The addition of PAM to glutathione, a series of S-alkylated glutathiones, and leukotriene C(4) results in the consumption of O(2) and the production of the corresponding amidated peptide and glyoxylate. This reaction proceeds in two steps with the intermediate formation of a C terminal alpha-hydroxyglycine-extended peptide. Amidated glutathione (gammaGlu Cys-amide) is a relatively poor substrate for glutathione S-transferase with a V/K value that is 1.3% of that for glutathione. Peptide substrates containing a penultimate hydrophobic or sulfur-containing amino acid exhibit the highest (V/K)(app) values for PAM-catalyzed amidation. The S-alkylated glutathiones incorporate both features in the penultimate position with S-decylglutathione having the highest (V/K)(app) of the substrates described in this report. PMID- 12646263 TI - Reversible denaturation of the soybean Kunitz trypsin inhibitor. AB - The soybean Kunitz trypsin inhibitor (SKTI) is a beta-sheet protein with unusual stability to chemical and thermal denaturation. Different spectroscopic criteria were used to follow the thermal denaturation and renaturation of SKTI. Upon heating to 70 degrees C, changes in UV difference spectra showed increased absorbance at 292 and 297 nm, attributable to perturbation of aromatic residues. Cooling the protein resulted in restoration of the native spectrum unless reduced with dithiothreitol. Far- and near-UV CD spectra also indicate thermal unfolding involving the core tryptophan and tyrosine residues. Both CD and UV-absorbance data suggest a two-state transition with the midpoint at approximately 65 degrees C. CD data along with the increased fluorescence intensity of the reporter fluorophore, 1-anilino-8-naphthalenesulfonate with SKTI, between 60 and 70 degrees C, are consistent with a transition of the native inhibitor to an alternate conformation with a more molten state. Even after heating to 90 degrees C, subsequent cooling of SKTI resulted in >90% of native trypsin inhibition potential. These results indicate that thermal denaturation of SKTI is readily reversible to the native form upon cooling and may provide a useful system for future protein folding studies in the class of disordered beta-sheet proteins. PMID- 12646262 TI - Loss of p53 induces M-phase retardation following G2 DNA damage checkpoint abrogation. AB - Most cell lines that lack functional p53 protein are arrested in the G2 phase of the cell cycle due to DNA damage. When the G2 checkpoint is abrogated, these cells are forced into mitotic catastrophe. A549 lung adenocarcinoma cells, in which p53 was eliminated with the HPV16 E6 gene, exhibited efficient arrest in the G2 phase when treated with adriamycin. Administration of caffeine to G2 arrested cells induced a drastic change in cell phenotype, the nature of which depended on the status of p53. Flow cytometric and microscopic observations revealed that cells that either contained or lacked p53 resumed their cell cycles and entered mitosis upon caffeine treatment. However, transit to the M phase was slower in p53-negative cells than in p53-positive cells. Consistent with these observations, CDK1 activity was maintained at high levels, along with stable cyclin B1, in p53-negative cells. The addition of butyrolactone I, which is an inhibitor of CDK1 and CDK2, to the p53-negative cells reduced the floating round cell population and induced the disappearance of cyclin B1. These results suggest a relationship between the p53 pathway and the ubiquitin-mediated degradation of mitotic cyclins and possible cross-talk between the G2-DNA damage checkpoint and the mitotic checkpoint. PMID- 12646264 TI - Mitochondrial targets of oxidative stress during renal ischemia/reperfusion. AB - Endogenous tyrosine nitration and inactivation of manganese superoxide dismutase (MnSOD) has previously been shown to occur in both human and rat chronic renal allograft rejection. To elucidate the time course of MnSOD inactivation and mitochondrial dysfunction at earlier times during renal transplantation, we developed a rodent model of renal ischemia/reperfusion (I/R). Renal function was significantly impaired at 16 h reperfusion following 30 min of warm ischemia. Tyrosine nitration of specific mitochondrial proteins, MnSOD and cytochrome c, occurred at the earliest time point examined, an event that preceded significant renal injury. Interestingly, a small percentage of both mitochondrial proteins were also located in the cytosol. This leakage and decreased adenosine 5(') triphosphate levels indicate loss of mitochondrial membrane integrity during renal I/R. Inactivation of MnSOD occurred rapidly in this model of renal I/R, suggesting that loss of MnSOD activity leads to further renal injury and nitration of other mitochondrial targets. PMID- 12646265 TI - Leukotriene B4 omega-side chain hydroxylation by CYP4F5 and CYP4F6. AB - Leukotriene B(4) (LTB(4)) is a lipid mediator that plays an important role in inflammation. Metabolism of LTB(4) by cytochrome P450 (CYP) enzymes belonging to the CYP4F subfamily is considered to be of importance for the regulation of inflammation. This study investigates LTB(4) metabolism by recombinant rat CYP4F5 and CYP4F6 expressed in a yeast system and by microsomes isolated from rat organs expressing CYP4F mRNA. CYP4F6 was found to convert LTB(4) into 19-hydoxy- and 18 hydroxy-LTB(4) with an apparent K(m) of 26 microM, and CYP4F5 was found to convert LTB(4) primarily into 18-hydroxy-LTB(4) with an apparent K(m) of 9.7 microM. The rate of formation of 18-hydroxy-LTB(4) by CYP4F5 was surprisingly high. At a substrate concentration of 30 microM, the rate of formation was about 15 nmol/min/mg microsomal protein, approximately 30 times faster than the reaction catalyzed by CYP4F6. Analysis of LTB(4) metabolism by microsomes isolated from various tissues from the rat suggests that CYP4F5 and CYP4F6 are active in the lung and to some extent in the brain, kidney, and testis. CYP4F5 and CYP4F6, due to their capacities to metabolize LTB(4), may play important roles in modulating inflammatory response in these organs. PMID- 12646266 TI - AGE-breakers cleave model compounds, but do not break Maillard crosslinks in skin and tail collagen from diabetic rats. AB - Advanced glycation end products (AGE), formed by nonenzymatic Maillard reactions between carbohydrate and protein, contribute to the increase in chemical modification and crosslinking of tissue proteins with age. Acceleration of AGE formation in collagen during hyperglycemia, with resultant effects on vascular elasticity and basement membrane permeability, is implicated in the pathogenesis of diabetic complications. AGE-breakers, such as N-phenacylthiazolium (PTB) and N phenacyl-4,5-dimethylthiazolium (PMT) halides, have been proposed as therapeutic agents for reversing the increase in protein crosslinking in aging and diabetes. We have confirmed that these compounds, as well as the AGE-inhibitor pyridoxamine (PM), cleave the model AGE crosslink, phenylpropanedione, and have studied the effects of these compounds in reversing the increased crosslinking of skin and tail collagen isolated from diabetic rats. Crosslinking of skin collagen, measured as the half-time for solubilization of collagen by pepsin in 0.5M acetic acid, was increased approximately 5-fold in diabetic, compared to nondiabetic rats. Crosslinking of tail tendon collagen, measured as insolubility in 0.05 N acetic acid, was increased approximately 10-fold. Collagen preparations were incubated in the presence or absence of AGE-breakers or PM in phosphate buffer, pH 7.4, for 24h at 37 degrees C. These treatments did not decrease the half-time for solubilization of diabetic skin collagen by pepsin or increase the acid solubility of diabetic tail tendon collagen. We conclude that, although AGE breakers and PM cleave model crosslinks, they do not significantly cleave AGE crosslinks formed in vivo in skin collagen of diabetic rats. PMID- 12646267 TI - Singlet oxygen quenching by dietary carotenoids in a model membrane environment. AB - The ability of several dietary carotenoids to quench singlet oxygen in a model membrane system (unilamellar DPPC liposomes) has been investigated. Singlet oxygen was generated in both the aqueous and the lipid phase, with quenching by a particular carotenoid independent of the site of generation. However, singlet oxygen quenching is dependent on the carotenoid incorporated; xanthophylls exhibit a marked reduction in efficiency compared to the hydrocarbon carotenoids. Lycopene and beta-carotene exhibit the fastest singlet oxygen quenching rate constants (2.3-2.5 x 10(9)M(-1)s(-1)) with lutein the least efficient (1.1 x 10(8)M(-1)s(-1)). The other carotenoids, astaxanthin and canthaxanthin, are intermediate. Zeaxanthin exhibits anomalous behavior, and singlet oxygen quenching decreases with increasing amounts of zeaxanthin, leading to nonlinear plots for the decay of singlet oxygen with zeaxanthin concentration. Such differences are discussed in terms of carotenoid structure and their influence on the properties of the lipid membrane. The formation of aggregates by the polar carotenoids is also proposed to be of significance in their ability to quench singlet oxygen. PMID- 12646268 TI - The trypanothione-thiol system in Trypanosoma cruzi as a key antioxidant mechanism against peroxynitrite-mediated cytotoxicity. AB - Peroxynitrite, the reaction product between superoxide (O(*2)) and nitric oxide (*NO), is a powerful oxidizing species that contributes to macrophage competence against pathogens. In this context, peroxynitrite appears to play an important role in controlling infection by Trypanosoma cruzi, the unicellular parasite responsible for Chagas disease. T. cruzi contains various enzyme systems for the decomposition of hydroperoxides, all of which involve the participation of the low-molecular-weight dithiol trypanothione (N(1),N(8) bis(glutathionyl)spermidine) as a critical redox partner. A large fraction of the trypanothione-dependent antioxidant capacity of T. cruzi is linked to the tryparedoxin-tryparedoxin peroxidase system which has critical protein thiol groups. In this report we demonstrate that dihydrotrypanothione is readily consumed during peroxynitrite challenge to cells to yield the corresponding trypanothione disulfide. On the other hand, glutathione, which is present in T. cruzi at lower concentrations than trypanothione, is consumed to a much lesser extent and mainly evolves to glutathione-protein mixed disulfides. The inhibition of glutathione biosynthesis by buthionine sulfoximine, which decreases glutathione concentration to 10% of control after 20 h, neither affects the concentration of dihydrotrypanothione nor sensitizes T. cruzi to peroxynitrite mediated cytotoxicity. On the other hand, pretreatment of T. cruzi with diamide, which leads to a significant depletion (>70%) of dihydrotrypanothione, largely increases the extent of cellular nitration and inhibition of cell growth caused by peroxynitrite. Altogether, our findings support a key protective role for dihydrotrypanothione and the trypanothione-dependent antioxidant system in T. cruzi against peroxynitrite, which may facilitate the survival of trypanosomes within the oxidative environment of activated macrophages. PMID- 12646269 TI - Electron transfer is activated by calmodulin in the flavin domain of human neuronal nitric oxide synthase. AB - The objective of this study was to clarify the mechanism of electron transfer in the human neuronal nitric oxide synthase (nNOS) flavin domain using the recombinant human nNOS flavin domains, the FAD/NADPH domain (contains FAD- and NADPH-binding sites), and the FAD/FMN domain (the flavin domain including a calmodulin-binding site). The reduction by NADPH of the two domains was studied by rapid-mixing, stopped-flow spectroscopy. For the FAD/NADPH domain, the results indicate that FAD is reduced by NADPH to generate the two-electron-reduced form (FADH(2)) and the reoxidation of the reduced FAD proceeds via a neutral (blue) semiquinone with molecular oxygen or ferricyanide, indicating that the reduced FAD is oxidized in two successive one-electron steps. The neutral (blue) semiquinone form, as an intermediate in the air-oxidation, was unstable in the presence of O(2). The purified FAD/NADPH domain prepared under our experimental conditions was activated by NADP(+) but not NAD(+). These results indicate that this domain exists in two states; an active state and a resting state, and the enzyme in the resting state can be activated by NADP(+). For the FAD/FMN domain, the reduction of the FAD-FMN pair of the oxidized enzyme with NADPH proceeded by both one-electron equivalent and two-electron equivalent mechanisms. The formation of semiquinones from the FAD-FMN pair was greatly increased in the presence of Ca(2+)/CaM. The air-stable semiquinone form, FAD-FMNH(.), was further rapidly reduced by NADPH with an increase at 520 nm, which is a characteristic peak of the FAD semiquinone. Results presented here indicate that intramolecular one-electron transfer from FAD to FMN is activated by the binding of Ca(2+)/CaM. PMID- 12646270 TI - Cloning, tissue distribution, and functional studies of a new cytochrome P450 3A subfamily member, CYP3A45, from rainbow trout (Oncorhynchus mykiss) intestinal ceca. AB - In trout and mammals, the major extrahepatic expression site for CYP3A forms is in the intestine. A cDNA encoding a new CYP3A subfamily member was isolated from rainbow trout intestinal ceca by reverse transcriptase-polymerase chain reaction (RT-PCR), followed by rapid amplification of cDNA ends (RACE)-PCR. In a set of two primers for PCR, a consensus sequence in the highly conserved regions in 17 CYP3A sequences was used for one primer, and the second primer was designed based on adapter sequence ligated on the 5(') and 3(') cDNA ends. The 3(') and 5(') end nucleotide sequences of RACE-PCR products were used for the priming sites for the full-length cDNA in RT-PCR. The resulting 2615-bp cDNA contained an open reading frame of 1554 bp encoding a 518-amino acid residue protein (M(r)=59057.13, pI=6.15) with 26 amino acid differences from that of the previously cloned rainbow trout CYP3A27. The cDNA was assigned as CYP3A45 by the P450 Nomenclature Committee. The deduced amino acid sequence of rainbow trout CYP3A45 was 94% identical with trout CYP3A27, 72% with killifish CYP3A56, and 71% with both medaka CYP3A40 and killifish CYP3A30 in positional alignment comparisons. Northern blotting by a CYP3A45-specific nucleotide probe showed that the major expression site was the intestinal ceca rather than the liver in both male and female trout. Recombinant baculovirus containing a CYP3A45 cDNA (Bv-3A45) was constructed under polyhedrin promoter of the Autographa californica nuclear polyhedrosis virus and used to express CYP3A45 protein in Spodoptera frugiperda cells. The Western blot showed that the expressed CYP3A45 protein comigrated with purified LMC5 P450 and was recognized by anti-LMC5 polyclonal antibodies. The expressed CYP3A45 showed catalytic activities for the 6 beta-, 2 beta-, and 16 beta-hydroxytestosterones of 1.76, 0.193, and 0.078 nmol/min/nmol CYP3A45, respectively. In summary, a second form of CYP3A with steroid hydroxylase activity, CYP3A45, has been cloned from rainbow trout and the major site of expression was in the intestinal tissues. PMID- 12646271 TI - Protection of epidermal cells against UVB injury by the antioxidant selenium containing single-chain Fv catalytic antibody. AB - The antioxidant effect of selenium-containing single-chain Fv catalytic antibody (Se-scFv2F3), a new mimic of glutathione peroxidase, was confirmed using a model system in which cultured rat skin epidermal cells were injured by ultraviolet B (UVB). The cell damage was characterized in terms of lipid peroxidation of the cells, cell viability, and cell membrane integrity. The injury effects of UVB and protection effects of Se-scFv2F3 on the cells were studied using the model system. UVB can damage the cells severely. Upon precultivation of the cells with 0.4U/ml Se-scFv2F3, however, the damage was significantly reduced as shown by the increase in cell viability, the decrease in the malondialdehyde and hydrogen peroxide levels, and the normalization of lactate dehydrogenase activity. In addition, a novel finding that Se-scFv2F3 can stimulate cultured epidermal cells to proliferate under certain conditions was observed. PMID- 12646272 TI - Mutational analysis of the mouse aryl hydrocarbon receptor tyrosine residues necessary for recognition of dioxin response elements. AB - Tyrosine phosphorylation of the aryl hydrocarbon receptor (AhR), a member of the basic helix-loop-helix/PER-ARNT-SIM transcription factor family, has been shown to regulate its dioxin response elements (DRE) binding ability, although no specific residues have been directly demonstrated to be phosphorylated. Of the 23 tyrosines in the mouse AhR, 19 are conserved across all mammalian species sequenced thus far. The studies presented here were conducted to examine tyrosine residue(s) that are both likely candidates of phosphorylation and necessary for DNA binding and/or transcriptional activity of the AhR. Two-dimensional gel electrophoresis of phosphatase-treated AhR indicated that the receptor is phosphorylated on serine/threonine and tyrosine residues. Computational analysis predicted several highly conserved tyrosine residues to be phosphorylated. Both the N terminus (amino acids 1-399) and the C terminus (amino acids 399-805) of the mouse receptor synthesized in vitro using a rabbit reticulocyte lysate system are tyrosine phosphorylated as detected by antiphosphotyrosine antibodies. Furthermore, the N-terminal AhR bound DRE in a ligand-dependent manner similar to that by the full-length receptor, suggesting that phosphorylated tyrosines involved in DNA binding are likely located in the region between residues 1 and 399. Mouse AhR tyrosine (Y) residues were evaluated by phenylalanine (F) mutational analysis for both DNA binding (electrophoretic mobility shift assays; EMSAs) and ability to induce a DRE-driven reporter gene in transiently transfected AhR-deficient cells. Of the 12 tyrosine residues in the N-terminal AhR, only a tyrosine 9 mutant (AhRY9F) significantly decreased DRE binding as determined by EMSA. Similarly, only the AhRY9F mutant decreased the DRE-driven luciferase expression in AhR-deficient cells. Overall, these data strongly suggest that the putative posttranslational modification at, or mediated by, tyrosine 9, and not any other individual mouse AhR tyrosine residue, is necessary for AhR DRE binding and transcriptional activity. PMID- 12646274 TI - Arginine 25 and Arginine 28 of lactoferrin are critical for effective heparin neutralization in blood. AB - The interaction of lactoferrin with endogenous heparin-like molecules modulates glycosaminoglycan-mediated biological processes. We performed site-specific mutagenesis and expressed recombinant lactoferrin and lactoferrin mutants by the baculovirus insect cell expression system. Five basic residues at the lactoferrin N terminus; Arg 5, Arg 25, Arg 28, Lys 29, and Arg 31, were individually replaced by alanines. Heparin chromatography on fast-performance liquid chromatography system showed that the NaCl concentrations corresponding to the peak of each eluted recombinant protein from the column were 665, 620, 540, 550, 630, or 650 mM for wild-type recombinant lactoferrin, Arg 5, Arg 25, Arg 28, Lys 29, or Arg 31 recombinant lactoferrin mutant, respectively. We compared the ability of each mutated lactoferrin derivative to neutralize glycosaminoglycans in the thrombin serpin inhibition assays. In comparison to wild-type recombinant lactoferrin, all the mutants showed decreased ability to neutralize glycosaminoglycan in a dose dependent manner. The mutations of lactoferrin at Arg 25 and Arg 28 demonstrated the most striking decrease in lactoferrin's ability to neutralize various glycosaminoglycans in both enzymatic and plasma clotting-based experiments. Therefore, our results suggest that Arg 25 and Arg 28 are the critical basic residues at the lactoferrin N terminus responsible for heparin binding. The other basic residues on the N terminus, Arg 5, Lys 29, and Arg 31, also contribute to heparin binding by presenting an additional cationic motif. PMID- 12646273 TI - Expression and purification of recombinant tristetraprolin that can bind to tumor necrosis factor-alpha mRNA and serve as a substrate for mitogen-activated protein kinases. AB - Tristetraprolin (TTP) is an mRNA-binding protein, but studies of this interaction have been difficult due to problems with the purification of recombinant TTP. In the present study, we expressed human and mouse TTP as glutathione S-transferase and maltose-binding protein (MBP) fusion proteins in Escherichia coli, and purified them by affinity resins and Mono Q chromatography. TTP cleaved from the fusion protein was identified by immunoblotting, MALDI-MS, and protein sequencing, and was further purified to homogeneity by continuous-elution SDS-gel electrophoresis. Purified recombinant TTP bound to the AU-rich element of tumor necrosis factor-alpha (TNFalpha) mRNA and this binding was dependent on Zn(2+). Results from sizing columns suggested that the active species might be in the form of an oligomer of MBP-TTP. Recombinant TTP was phosphorylated by three members of the mitogen-activated protein (MAP) kinase family, p42, p38, and JNK, with half-maximal phosphorylation occurring at approximately 0.5, 0.25, and 0.25 microM protein, respectively. Phosphorylation by these kinases did not appear to affect the ability of TTP to bind to TNFalpha mRNA under the assay conditions. This study describes a procedure for purifying nonfusion protein TTP to homogeneity, demonstrates that TTP's RNA-binding activity is zinc dependent, and that TTP can be phosphorylated by JNK as well as by the other members of the greater MAP kinase family. PMID- 12646275 TI - Light-induced increase in free Mg2+ concentration in spinach chloroplasts: measurement of free Mg2+ by using a fluorescent probe and necessity of stromal alkalinization. AB - Free Mg(2+) in chloroplasts may contribute to the regulation of photosynthetic enzymes, but adequate methodology for the determination of free Mg(2+) concentration ([Mg(2+)]) in chloroplasts has been lacking. We measured internal chloroplast [Mg(2+)] by using a Mg-sensitive fluorescent indicator, mag-fura-2. In intact, dark-kept spinach chloroplasts, internal [Mg(2+)] was estimated to be 0.50 mM, and illumination caused an increase in [Mg(2+)] to 2.0mM in the stroma. The light-induced increase in [Mg(2+)] was inhibited by a blocker of driven electron transport and uncouplers. The K(+)-specific ionophore valinomycin inhibited the [Mg(2+)] increase in the absence of external K(+), and addition of KCl restored the [Mg(2+)] increase. NH(4)Cl, which induces stromal alkalinization, enhanced the [Mg(2+)] increase. A Ca(2+)-channel blocker, ruthenium red, inhibited the [Mg(2+)] increase, but LaCl(3) had no effect. These results indicate that stromal alkalinization is essential for light-induced increase in [Mg(2+)]. This system for measuring internal chloroplast [Mg(2+)] might provide a suitable system for assay of Mg(2+) transport activity of chloroplast membranes. PMID- 12646276 TI - Wide distribution of cysteine-rich secretory proteins in snake venoms: isolation and cloning of novel snake venom cysteine-rich secretory proteins. AB - Cysteine-rich secretory proteins (CRISPs) are found in epididymis and granules of mammals, and they are thought to function in sperm maturation and in the immune system. Recently, we isolated and obtained clones for novel snake venom proteins that are classified as CRISP family proteins. To elucidate the distribution of snake venom CRISP family proteins, we evaluated a wide range of venoms for immuno cross-reactivity. Then we isolated, characterized, and cloned genes for three novel CRISP family proteins (piscivorin, ophanin, and catrin) from the venom of eastern cottonmouth (Agkistrodon piscivorus piscivorus), king cobra (Ophiophagus hannah), and western diamondback rattlesnake (Crotalus atrox). Our results show the wide distribution of snake venom CRISP family proteins among Viperidae and Elapidae from different continents, indicating that CRISP family proteins compose a new group of snake venom proteins. PMID- 12646278 TI - Cytochrome b5 reductase and cytochrome b5 support the CYP2E1-mediated activation of nitrosamines in a recombinant Ames test. AB - With CYP2E1 in vitro both the first and the second electron of the catalytic cycle can come from cytochrome b(5) via either NADPH-cytochrome P450 reductase or NADH-cytochrome b(5) reductase, and the presence of cytochrome b(5) stimulates CYP2E1 turnover both in vitro and in vivo. To determine whether electron input via the NADH-dependent pathway was similarly functional in whole cells and necessary for the stimulation by cytochrome b(5), we constructed five plasmids designed to express human CYP2E1 in various combinations with cytochrome b(5) reductase, cytochrome b(5), and cytochrome P450 reductase. CYP2E1 activity in Salmonella typhimurium cells transformed with each plasmid was assessed by mutagenic reversion frequency in the presence of dimethylnitrosamine. A fivefold increase in reversion frequency when cytochrome b(5) was coexpressed with P450 reductase was abolished by disruption of heme-binding in cytochrome b(5) by site directed mutagenesis (His68Ala), suggesting that electron transfer to cytochrome b(5) was necessary for the stimulation. Addition of cytochrome b(5) reductase to the cytochrome b(5)/P450 reductase coexpression plasmid did not further increase the stimulation by cytochrome b(5), but b(5) reductase could support CYP2E1 activity in the absence of P450 reductase at a level equivalent to that obtained with just CYP2E1 and P450 reductase. Neither cytochrome b(5) reductase nor cytochrome b(5) alone could support CYP2E1 activity. These results demonstrate that the cytochrome b(5) reductase/cytochrome b(5) pathway can support CYP2E1 activity in bacterial cells. PMID- 12646277 TI - Binding of fatty acids to the uncoupling protein from brown adipose tissue mitochondria. AB - The uncoupling protein 1 (UCP1) is a H(+) carrier which plays a key role in heat generation in brown adipose tissue. The H(+) transport activity of UCP1 is activated by long-chain fatty acids and inhibited by purine nucleotides. While nucleotide binding has been well characterized, the interaction of fatty acid with UCP1 remains unknown. Here I demonstrate the binding of fatty acids by competition with a fluorescent nucleotide probe 2(')-O-dansyl guanosine 5(') triphosphate (GTP), which has been shown previously to bind at the nucleotide binding site in UCP1. Fatty acids but not their esters competitively inhibit the binding of 2(')-O-dansyl GTP to UCP1. The fatty acid effect was enhanced at higher pH, suggesting the binding of fatty acid anion to UCP1. The inhibition constants K(i) were determined by fluorescence titrations for various fatty acids. Short-chain (C<8) fatty acids display no affinity, whereas medium-chain (C10-14) and unsaturated C18 fatty acids exhibit stronger affinity (K(i)=65 microM, for elaidic acid). This specificity profile agrees with previous functional data obtained in both proteoliposomes and mitochondria, suggesting a possible physiological role of this fatty acid binding site. PMID- 12646279 TI - Acute activation of the spinal cord metabotropic glutamate subtype-5 receptor leads to cold hypersensitivity in the rat. AB - Activation of spinal cord dorsal horn ionotropic glutamate receptors leads to pain-related behaviors. However, the role of spinal metabotropic glutamate receptors (mGlu), particularly the mGlu5 receptor subtype, in nociception has not been well characterized. A recently described subtype selective and potent mGluR5 antagonist, 2-methyl-6-(phenylethynyl)pyridine (MPEP) was used to evaluate the role of the mGlu5 receptor in cold sensitivity. Intrathecal (i.t.) injection of group I (mGlu1 and mGlu5 receptors) mGlu receptor-selective agonist (RS)-3,5 dihydroxyphenylglycine (DHPG) increased the hind paw frequency and duration of lifting of rats placed on a cold (4 degrees C) surface, a behavior similarly observed in rats with a chronic constriction injury (CCI) of the sciatic nerve. In contrast, rats i.t. injected with DHPG did not display increased lifting when placed on a room temperature surface. I.t. injection of MPEP before i.t. injection of DHPG blocked DHPG-evoked cold hypersensitivity, suggesting that activation of spinal mGlu5 receptors induces this behavioral response. In contrast, i.t. injection of MPEP after i.t. injection of DHPG had no effect. In addition, i.t. injection of MPEP did not affect cold hypersensitivity in rats with a CCI. These data suggest that acute activation of spinal cord mGlu5 receptors results in increased sensitivity to cold, but ongoing cold hypersensitivity does not involve activation of the mGlu5 receptor. PMID- 12646280 TI - The GABA(A) receptor antagonist picrotoxin inhibits 5-hydroxytryptamine type 3A receptors. AB - For a number of years it has been known that the CNS convulsant picrotoxin inhibits the GABA(A) receptor, an anion-selective member of the ligand-gated ion channel (LGIC) superfamily. PTX also inhibits other anion-selective LGIC members, such as GABA(C), glycine and glutamate-gated Cl(-) channels. In the present report, we tested the ability of picrotoxin to inhibit cation-selective 5-HT(3A) receptors. Murine 5-HT(3A) receptors were expressed in HEK293 cells, and functionally evaluated using whole-cell patch clamp recording. Picrotoxin inhibited 5-HT-gated currents in a concentration-dependent manner, with an IC(50) of approximately 30 microM. Moreover, the blockade by PTX was non-competitive and use-facilitated. Pentylenetetrazole and U-93631, ligands that act at a domain similar to that of picrotoxin in GABA(A) receptors, also inhibited the 5-HT(3A) receptor. For each ligand tested, its potency was 5-10 fold lower than typically observed in GABA(A) receptors. Our results demonstrate that, in addition to being a relatively non-selective inhibitor of anionic LGICs, picrotoxin also inhibits the cation-selective 5-HT(3A) receptor. Moreover, the fact that both PTZ and U 93631 similarly inhibit the 5-HT(3A) receptor is consistent with the suggestion that the site of picrotoxin action in this receptor may be comparable to that in anion-selective LGICs. PMID- 12646282 TI - Differential effect of dietary selenium on the long-term neurotoxicity induced by MDMA in mice and rats. AB - We examined the effect of dietary selenium (Se) on the long-term effect of 3,4 methylenedioxymethamphetamine (MDMA) on dopamine (DA) and 5-hydroxytryptamine (5 HT) containing neurons in the brain of mice and rats. Animals were fed either a Se-deficient (<0.02 ppm) or Se-replete (0.2 ppm) diet for 8 weeks. On the seventh week mice received three injections of MDMA (15 mg/kg, i.p. 3 h apart) or saline and rats a single dose of MDMA (12.5 mg/kg i.p.) or saline. All animals were sacrificed 7 days later. MDMA administration to mice depleted striatal DA concentration in both dietary groups, although depletion was considerably larger in the Se-deficient mice (64%) than Se-replete mice (30%). In addition, a decrease in 5-HT (17-32%) occurred in brain regions of Se-deficient but not Se replete mice. In rats, MDMA decreased cortical [(3)H]-paroxetine binding (62%) and 5-HT content, the depletion being similar in the Se-deficient and Se-replete groups. No DA loss occurred in either group. There was no difference in the hyperthermic response induced by MDMA in Se-deficient or Se-replete animals. The Se-deficient diet decreased glutathione peroxidase (GPx) activity by 30% in mouse striatum and cortex and increased the degree of lipid peroxidation in cortical synaptosomes. Se-deficient rats also showed a decrease in brain GPx activity compared with the Se-replete group, but the degree of lipid peroxidation in synaptosomes was similar in both dietary groups. These results suggest that the antioxidant capacity of rats and mice differ leading to a differential susceptibility to the oxidative stress caused by MDMA in situations of low dietary Se. PMID- 12646281 TI - Reduced social interaction following 3,4-methylenedioxymethamphetamine is not associated with enhanced 5-HT 2C receptor responsivity. AB - This study examined the long-term change in serotonergic (5-hydroxytryptamine, 5 HT) neuronal function and 5-HT(2C) receptor agonist-induced behaviour following treatment of young rats with 3,4-methylenedioxymethamphetamine (MDMA). On post natal day (PND) 28, Lister-hooded rats received either MDMA (15 mg/kg i.p.) or saline (1 ml/kg i.p.) twice daily for 3 days. On PND 50 social interaction was assessed between treatment-matched pairs of rats derived from separate litters. The effect of either the 5-HT(2C) receptor agonist, m-chlorophenylpiperazine (m CPP, 2.5 or 1 mg/kg i.p., respectively) or saline was examined on open-field exploration (PND 52) and elevated plus-maze behaviour (PND 56). Acutely, MDMA produced hyperlocomotion and hypothermia compared with saline injection (p<0.001). Following 20 days abstinence, social interaction was decreased by 26% (p<0.05) in MDMA pre-treated rats compared with saline controls, without any change in locomotion. There was no difference in open-field or elevated plus-maze behaviour between pre-treatment groups. m-CPP caused hypolocomotion in the open field and decreased both the percentage entries into, and time spent in, the open arms of the elevated plus-maze to a comparable extent in MDMA and saline pre treated rats. Hippocampal and frontal cortical 5-HT and 5-hydroxyindoleacetic acid (5-HIAA) were significantly reduced in MDMA pre-treated rats, without any change in [(3)H]paroxetine binding or plasma corticosterone levels. These data suggest that the MDMA-induced reduction in social interaction is not mediated via alteration of 5-HT(2C) receptor function. PMID- 12646283 TI - Olanzapine and clozapine but not haloperidol reverse subchronic phencyclidine induced functional hyperactivity of N-methyl-D-aspartate receptors in pyramidal cells of the rat medial prefrontal cortex. AB - In the present study, we have investigated and compared the ability of olanzapine, clozapine and haloperidol to modulate phencyclidine (PCP)-induced effect in pyramidal cells of the medial prefrontal cortex (mPFC) of rats using the techniques of intracellular recording and voltage-clamp. Subchronic treatment of rats with PCP (2 mg/kg, b.i.d., 7 days, 48-60 h withdrawal) produced: (1) a depolarized resting membrane potential, a decrease of slow after hyperpolarization (sAHP) and spike frequency adaptation, (2) a shift of the concentration response curve of N-methyl-D-aspartate (NMDA), but not (+/-)-alpha amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA), to the left, (3) a decrease of the paired pulse facilitation (PPF) with an increase of excitatory postsynaptic current variance (EPSC variance), and (4) a reduction of the blockade of NMDA response by in vitro application of PCP. Repeated treatment with either olanzapine or clozapine, but not haloperidol, completely prevented the aforementioned subchronic PCP-induced effects. The present results indicate that the atypical antipsychotic drugs (APDs) clozapine and olanzapine share a common property in preventing subchronic PCP-induced functional hyperactivity of NMDA receptors. PMID- 12646284 TI - RO-10-5824 is a selective dopamine D4 receptor agonist that increases novel object exploration in C57 mice. AB - Novelty seeking as a behavioral phenomenon emerges as a compromise between approach and avoidance behavior. Although novelty seeking is thought to play a role in drug abuse and in cognition, the biological basis for this construct is poorly understood. At a genetic level, dopamine D4 receptors (D4R) appear to be critical for the behavioral expression of novelty seeking. In humans, polymorphisms of D4R have been associated with novelty-seeking traits in general and attention deficit-hyperactivity disorder in particular. Similarly, D4R (-/-) mice exhibit less novel object exploration than D4R (+/+) mice. Due to of the paucity of selective D4R ligands for use in behavioral pharmacology studies, few studies have examined the behavioral effects of D4R compounds in animals. The present experiments characterized RO-10-5824, a new, selective D4R partial agonist with minimal affinity for dopamine D2 and D3 receptors, and tested the hypothesis that activation of D4R increases the investigation by mice of a novel object placed in the center of a familiar open field. C57BL/6J and DBA/1J male mice were used in a dose response study of the selective D4R partial agonist RO 10-5824 (0, 1.0, 3.0, or 10.0 mg/kg). While having no effect on the amount of locomotor activity in novel or familiar environments, RO-10-5824 (10.0 mg/kg) increased time spent in the center of the enclosure in the presence of a novel object in C57 but not DBA mice. These results support the hypothesis that stimulation of D4R can enhance novelty seeking in mice and that this effect may be dependent on subtle genetic differences. PMID- 12646285 TI - Mu-opioid receptor activation prevents apoptosis following serum withdrawal in differentiated SH-SY5Y cells and cortical neurons via phosphatidylinositol 3 kinase. AB - Opioid peptides and alkaloids exert their effects via G protein-coupled receptors (GPCRs). It has been shown that, in addition to trophic factors, some GPCRs are able to activate the phosphatidylinositol 3-kinase/Akt (PI 3-K/Akt) signal transduction pathway, thus leading to cell survival. The aim of this study was to test whether activation of mu-opioid receptors has protective effects on serum withdrawal-induced cell death and to study the possible implication of PI 3-K in this process. In SH-SY5Y neuroblastoma cells fully differentiated by exposure to retinoic acid for five days, the enkephalin derivative selective mu-agonist DAMGO (0.1-2 microM) and the alkaloid morphine (0.1-10 microM) promoted cell survival after serum deprivation (MTT and trypan blue exclusion assays), without inducing cell proliferation. These effects were fully reversed by naloxone, by the selective mu-antagonist beta-funaltrexamine (beta-FNA) and also by the specific PI 3-K inhibitor LY294002. The two agonists stimulated Akt phosphorylation and the effect was also abolished by beta-FNA and by LY294002. In mouse primary cortical neurons, DAMGO reduced the percentage of apoptosis after 6, 12, 24 and 48 h of serum withdrawal; as determined by Hoechst staining. This effect was blocked by beta-FNA, by pre-treatment with pertussis toxin and by LY294002. DAMGO also stimulated Akt phosphorylation via PI 3-K in this primary neuronal culture. Together, these results indicate that stimulation of the mu-opioid receptor promotes neuronal survival in a G(i/o)-linked, PI 3-K-dependent signaling cascade and suggest that Akt may be a key downstream kinase involved in this anti apoptotic effect. PMID- 12646286 TI - Protein kinase C epsilon is involved in ethanol potentiation of glycine-gated Cl( ) current in rat neurons of ventral tegmental area. AB - Previously, we demonstrated that ethanol potentiates glycine current (I(Gly)) in 35% of neurons freshly isolated from the ventral tegmental area (VTA) of rats (J. Pharmacol. Exp. Ther. 296 (2001) 77). In the present study, we examined the role of protein kinase C (PKC) in this action of ethanol on VTA neurons from young rats. Extracellular ethanol and intracellular ATP-gamma-S when applied separately potentiated I(Gly). However, ethanol potentiation of I(Gly) was significantly reduced in neurons dialyzed with 2 mM ATP-gamma-S. Phorbol-12-myristate-13 acetate (PMA, 10 nM), a PKC activator also increased I(Gly) and reduced ethanol potentiation of I(Gly). In addition, GF109203X (0.2 microM), a PKC inhibitor antagonized the potentiation effects produced either by PMA or by ethanol. Thus, ethanol potentiation of I(Gly) may be associated with PKC activation. While intracellular application of 1,2-bis(aminophenoxy)-ethane-N,N,N,N'-tetraacetic acid, a Ca(2+) chelator or Go6976, an inhibitor of Ca(2+)-dependent PKC had no appreciable effect on ethanol potentiation of I(Gly), translocation inhibitor peptide (PKC(epsilon)-TIP) (500 nM) significantly reduced ethanol potentiation, an action the translocation inhibitor peptide negative control (PKC(epsilon)-TIP NC) (500 nM) did not have. These results suggest that the activation of PKC(epsilon) isoenzyme contributes to ethanol-induced potentiation of GlyR function. PMID- 12646287 TI - Halogenated cytisine derivatives as agonists at human neuronal nicotinic acetylcholine receptor subtypes. AB - Cytisine (cy) is a potent and competitive partial agonist at alpha4 subunit containing nicotinic acetylcholine (nACh) receptors while at homomeric alpha7 nACh receptors it behaves as a full agonist with a relatively lower potency. In the present study, we assessed the effects of bromination or iodination of the pyridone ring of cy and N-methylcytisine (N-Me-cy) on the effects of these compounds on recombinant human (h) alpha7, halpha4beta2 and halpha4beta4 nACh receptors expressed in clonal cell lines and Xenopus oocytes. Halogenation at C(3) of cy or N-Me-cy usually brings about a marked increase in both affinity and efficacy at halpha7, halpha4beta2 and halpha4beta4 nACh, the extent of which depends on whether the halogen is bromine or iodine, and upon receptor subtype. The effects of halogenation at C(5) are strongly influenced by the specific halogen substituent so that bromination causes a decrease in both affinity and efficacy while iodination decreases affinity but its effects on efficacy range from a decrease (halpha7, halpha4beta4 nACh receptors) to a marked increase (halpha4beta2 nACh receptors). Based on these findings, which differ from those showing that neither the affinity nor efficacy of nicotine, 3-(2 azetidinylmethoxy)-pyridine or epibatidine are greatly affected by halogenation, dehalogenation or halogen exchange at equivalent positions, we suggest that cy, N Me-cy and their halo-isosteres bind to neuronal nACh receptors in a different orientation allowing the halogen atom to interact with a hydrophobic halogen accepting region within the predominantly hydrophobic agonist-binding pocket of the receptors. PMID- 12646288 TI - The substance P (NK1) receptor antagonist L-760735 inhibits fear conditioning in gerbils. AB - The ability of the substance P (NK(1) receptor) antagonist (SPA) L-760735 to inhibit conditioned fear was assessed in gerbils using a four plate apparatus. Animals that had been treated with diazepam (3 mg/kg) or L-760735 (3 mg/kg) 30 min before a 3 min conditioning session in the apparatus exhibited a release of plate crossings during the retest session approximately 3 h later. Plate crossings were also increased when animals received diazepam or L-760735 30 min before the retest session. In contrast, fluoxetine and venlafaxine (30 mg/kg) did not exhibit anxiolytic-like effects. During the retest session, gerbils drummed their hind feet on the floor; this behaviour was not observed spontaneously in gerbils that were naive to the apparatus. Foot drumming was abolished by pretreatment with L-760735 or diazepam (3 mg/kg) but was markedly increased following administration of fluoxetine or venlafaxine (30 mg/kg). Foot drumming elicited by aversive conditioning alone or in combination with fluoxetine was abolished by administration of L-760735 and by amygdala lesions involving the basolateral and lateral nuclei, indicating that this behaviour is an alarm signal or fear response mediated via release of substance P in brain circuits involving the amygdala. The observations provide further evidence for an anxiolytic-like profile of SPAs in preclinical assays and demonstrate a clear difference between the actions of SPAs and established antidepressant drugs. PMID- 12646289 TI - Endogenous CCK depresses contractile activity within the ascending myenteric reflex pathway of rat ileum. AB - The ascending excitatory reflex is an important part of the myenteric reflex. In order to study the ascending neural pathways, isolated segments of rat ileum were stimulated by electrical stimulation of the gut wall (20 V, 3 pulses per second, 1 ms) using platinum electrodes. The excitatory contractile response was recorded using perfused manometric side-hole tubing located 2 and 4 cm orally to the stimulation site. The contractile response to electrical stimulation was abolished by atropine (10(-6) M) or hexamethonium (10(-4) M). The excitatory response increased after administration of the cholecystokinin A (CCK(A)) receptor antagonists lorglumide (3x10(-6) M: +44.1%), devazepide (10(-8) M: +19.4%; 10(-7) M: +30.0%) and SR-27897 (10(-10) M: +21.8%, 10(-8) M: +47.0%, P<0.05, n=8). However, the CCK(B) receptor antagonist L-365,260 also caused a significant increase in the oral excitation (10(-6) M: +27.4%). sCCK-8 caused a significant reduction in the ascending response (10(-8) M: -11.5%) and induced spontaneously occurring contractions at doses ranging from 10(-10)-10(-6) M. CCK 9 significantly increased the ascending response (10(-7) M: +10.9%, P<0.05). However, caerulein (10(-10) M: -25.9%, 10(-8) M: -26.8%; P<0.01) and pentagastrin (10(-10) M: -20.2%, P<0.05; 10(-8) M: -23.7%, P<0.01; 10(-6) M: -28.3%, P<0.001) reduced the ascending contractile response significantly. These data, obtained with potent and highly specific CCK receptor antagonists, demonstrate an inhibitory role of endogenously released CCK within the ascending neural pathway. The data further suggest that exogenously applied CCK-related peptides have different effects on the myenteric reflex which might be due to excitation of the different involved neurons (short and long ascending inter- and motorneurons) in an unphysiological order. Thus in experiments investigating more complex neuronal circuits, experiments with antagonists should be regarded as more specific. PMID- 12646290 TI - Post-ischemic administration [correction of administeration] but not pre-ischemic administration [correction of administeration] of NG-nitro-L-arginine prevents spatial memory impairments and apoptosis by an inhibition of a delayed increase in NOx- in the hippocampus following repeated cerebral ischemia. AB - In the present study, we investigated the effects of N(G)-nitro-L-arginine (L NAME), an inhibitor of nitric oxide synthase, on repeated cerebral ischemia induced impairment of spatial memory of the 8-arm radial maze in rats. Repeated ischemia (10 min ischemia x 2 times with 1 h interval) impaired the spatial memory in the 8-arm radial maze test and produced apoptosis in the hippocampus 7 days after final occlusion, and gradually increased the NO(x)(-) levels approximately 30-180 min after the second reperfusion. Post-ischemic administration of L-NAME at a dose of 50 mg/kg, i.p. 30 min following the second occlusion, significantly attenuated the repeated ischemia-induced impairment of spatial memory in the 8-arm radial maze test and suppressed apoptosis in the hippocampus, and also significantly suppressed a delayed increase in the NO(x)(-) levels induced by repeated ischemia. However, pre-ischemic administration of L NAME at a dose of 50 mg/kg, i.p. 30 min before the first occlusion, caused about 90% mortality (the mortality rate of vehicle-treated group was 10%). These results suggest that the delayed generation of NO(x)(-) may cause spatial memory impairment and induction of apoptosis in the hippocampus in rats subjected to repeated ischemia. PMID- 12646291 TI - Ameliorative effect of NC-1900, a new AVP4-9 analog, through vasopressin V1A receptor on scopolamine-induced impairments of spatial memory in the eight-arm radial maze. AB - The mechanism by which NC-1900, a new pGlu-Asn-Cys(Cys)-Pro-Arg-Gly-NH(2) (AVP(4 9)) analog, improves spatial memory in rats using an eight-arm radial maze was examined. Even at very low doses (0.2 ng/kg for s.c., 1 microg/kg for p.o., 1 fg for i.c.v.) NC-1900 improved scopolamine-induced impairment of spatial memory. NC 1900 (1 ng/kg, s.c.) also improved impairment of spatial memory induced by pirenzepine, a muscarinic(1) (M(1)) receptor antagonist, and by KN-62, a Ca2+/calmodulin (CaM)-dependent protein kinase II inhibitor. [Pmp(1), Tyr(Me)(2)] Arg(8)-vasopressin, a vasopressin(1A) (V(1A)) receptor antagonist, and nicardipine, L-type Ca2+ blocker, but not OPC-31260, a V(2) antagonist, suppressed the effect of NC-1900 on scopolamine-induced impairment of spatial memory. A microdialysis study showed that NC-1900 did not affect acetylcholine release in the ventral hippocampus (VH) of intact rats or of scopolamine-treated rats. NC-1900 (1 microM) increased [Ca2+](i) in the VH than in the dorsal hippocampus (DH). Pretreatment with nicardipine (1 microM) and Ca2+ -free conditions inhibited the NC-1900-induced [Ca2+](i) response in the VH. Whereas co administration of NC-1900 (1 microM) and carbachol (500 microM) increased [Ca2+](i) in the VH. Moreover, nicardipine concentration-dependently inhibited the increase in [Ca2+](i) induced by the co-administration of NC-1900 and carbachol in the VH. These results suggest that NC-1900 activates the V(1A) receptor at the postsynaptic cholinergic nerve, and causes a transient influx of intracellular Ca2+ through L-type Ca2+ channels, to interact with the M(1) receptor. The activation of these Ca2+ -dependent processes induced by NC-1900 may be involved in the positive effect of NC-1900 on scopolamine-induced impairment of spatial memory. PMID- 12646293 TI - Women and depression: a millennial perspective. PMID- 12646294 TI - Epidemiology of women and depression. AB - BACKGROUND: Depression is the leading cause of disease-related disability among women in the world today. Depression is much more common among women than men, with female/male risk ratios roughly 2:1. AIMS: Recent epidemiological research is reviewed. Implications are suggested for needed future research. RESULTS: The higher prevalence of depression among women than men is due to higher risk of first onset, not to differential persistence or recurrence. Although the gender difference first emerges in puberty, other experiences related to changes in sex hormones (pregnancy, menopause, use of oral contraceptives, and use of hormone replacement therapy) do not significantly influence major depression. These observations suggest that the key to understanding the higher rates of depression among women than men lies in an investigation of the joint effects of biological vulnerabilities and environmental provoking experiences. CONCLUSIONS: Advancing understanding of female depression will require future epidemiologic research to focus on first onsets and to follow incident cohorts of young people through the pubertal transition into young adulthood with fine-grained measures of both sex hormones and gender-related environmental experiences. Experimental interventions aimed at primary prevention by jointly manipulating putative biological and environmental risk factors will likely be needed to adjudicate between contending causal hypotheses regarding the separate and joint effects of interrelated risk factors. PMID- 12646295 TI - The economics of women and depression: an employer's perspective. AB - BACKGROUND: While there is a growing literature on various aspects of depression in women, there is little research about the economic cost of depression in women. This analysis focuses on the direct and indirect cost to employers of female compared to male employees treated for depression, and their service utilization patterns. METHODS: We used a claims database from a national, Fortune 100 company to analyze the direct (medical and prescription drug) and indirect (disability and illness-related work absence) costs to an employer for female and male beneficiaries with depression. RESULTS: In 1998, the average female employee with depression cost this company $9265 compared to $8502 for male employees with depression. These women had significantly greater work absence costs which led to higher total costs than men, even though their medical costs were lower than those of comparable men. LIMITATIONS: Our analysis of the indirect costs associated with depressed female and male employees is limited to the costs of disability and sporadic illness-related work absences. The data available from this one employer did not allow accounting for the cost of reduced productivity while at work. CONCLUSIONS: We recommend that employers consider programs to improve the management of individuals with depression, in particular women. Also, further research is necessary to encourage the medical community to be more sensitive to the symptoms of depression in women. PMID- 12646296 TI - Cost-effectiveness of a primary care intervention for depressed females. AB - BACKGROUND: To date, there is little information about the differential impact of primary care interventions by gender. We conducted an exploratory cost effectiveness analysis by gender of an intervention to improve recognition and guideline-concordant treatment of depression in primary care. METHODS: Primary care practices that did not employ an onsite mental healthcare specialist were randomized to enhanced (intervention) versus usual care. All subjects met study criteria for current major depression. Medical Outcomes Study SF-36 scores were converted into quality-adjusted life years (QALYs) to compare the 1-year effectiveness of enhanced versus usual care by gender. Based on results of previous studies, antidepressant acceptors beginning a new depression treatment episode were the focus of the analysis. Statistical analyses included multivariate regression models controlling for sociodemographic and clinical covariates. RESULTS: In the main analysis, enhanced care for females was more expensive and more effective than usual care, at an additional cost of $5244 per QALY. For males, enhanced care was essentially cost and outcome neutral compared to usual care. The cost-effectiveness ratio estimates were robust to sensitivity analyses. Psychological side effects to the intervention may partially explain the limited effect of the intervention on outcomes for males. LIMITATIONS: We consider these results exploratory because the SF-36 to quality-adjusted life year conversion formula is preliminary and because of the relatively small sample size. CONCLUSIONS: The estimated cost-effectiveness ratio of this depression intervention is within the acceptable range for females, but not males. If replicated, these exploratory findings suggest that interventions to improve primary care depression treatment may need to be modified to improve their effectiveness in males while maintaining their effectiveness in females. PMID- 12646297 TI - Developmental changes in the phenomenology of depression in girls compared to boys from childhood onward. AB - BACKGROUND: Although it is well documented that teenage girls are at increased risk for depression, little is known about the importance of sex and development subsequent to onset of depression. In this article, we therefore report on sex differences in the developmental phenomenology of depression in a clinical sample. METHODS: Longitudinal analyses were used to examine changes in the risk of depression, patterns of comorbid diagnoses, and depressive symptoms from ages 8-13 years up to young adulthood (age 21) among 87 patients. RESULTS: Girls and boys were at similar risk for recurrent depression during follow-up. As girls got older, they had higher rates of comorbid eating disorders and lower rates of externalizing and substance use disorders than did boys; high risk periods for comorbid conditions also differed by sex. Comorbid disorders were usually contemporaneous with depression among girls but not among boys, and comorbidity patterns were influenced by age at depression onset. Girls with earlier (compared to later) onset depression were at lower risk for nonaffective disorders; boys evidenced the opposite pattern and were at particularly high risk for substance use disorders. There also were sex differences in developmental symptom patterns. LIMITATIONS: Results based on this initially clinically referred sample may not generalize to youths in other settings. CONCLUSIONS: Among depressed youths, sex associated developmental trajectories in comorbid disorders and depressive symptoms have implications for intervention. Such information may help identify optimal timing of efforts to reduce the risk of or to treat comorbid disorders, and help select developmentally appropriate target symptoms for girls versus boys. PMID- 12646298 TI - Interpersonal stress and depression in women. AB - The article presents a review and discussion of several aspects of the interpersonal context in which depression occurs that are unique to women. Women commonly experience depression in response to interpersonal life events, and also they contribute to the occurrence of stressful events and life contexts. Four key topics are reviewed: childrearing and parenting; romantic and marital relationships; generation of stressful life events; enduring social dysfunction even when not depressed. It is argued that depressed women are often locked into maladaptive interpersonal environments that contribute to the recurrence or chronicity of depression. Implications for treatment, conceptualization, and future research are noted. PMID- 12646299 TI - Personality and depression in women. AB - Personality disorders and traits should be assessed in studies concerned with the etiology and treatment of depression in women. There has been a considerable amount of research concerned with the effect of personality functioning on the etiology and treatment of depression, and much of this research has concerned personality traits and disorders for which significant gender differences and issues apply. The importance of personality functioning, and the particular relevance to women and gender-related issues, is illustrated in this paper with regard to dependent personality disorder and dependent personality traits. Implications for future research on the etiology and treatment of depression in women are provided. PMID- 12646300 TI - Hormones and mood: from menarche to menopause and beyond. AB - The lifetime prevalence of mood disorders in women is approximately twice that of men. The underlying causality of this gender difference is not yet understood. There is increasing scientific attention to the modulation of the neuroendocrine system by fluctuating gonadal hormones. This review attempts to summarize our current state of knowledge on the role and potential relevance of estrogen and other sex steroids to psychiatric disorders specific to women from menarche to menopause. The sudden appearance of higher levels of estrogen in puberty alters the sensitivity of the neurotransmitter systems. Moreover, the constant flux of estrogen and progesterone levels throughout the reproductive years portends constant modification of the neurotransmitter systems. Premenstrual syndromes may be the result of an altered activity or sensitivity of certain neurotransmitter systems. Pregnancy and delivery produce dramatic changes in estrogen and progesterone levels as well as significant suppression along the HPA axis, possibly increasing vulnerability to depression. At menopause, estrogen levels decline while pituitary LH and FSH levels increase. The loss of modulating effects of estrogen and progesterone may underlie the development of perimenopausal mood disorders in vulnerable women. The pattern of neuroendocrine events related to female reproduction is vulnerable to change and is sensitive to psychosocial, environmental, and physiological factors. Further research is needed to be able to identify specific genetic markers which might help us better understand how the balance between estrogen, progesterone, testosterone, and other steroid hormones affect neurotransmitter function. PMID- 12646302 TI - Pilocarpine-induced status epilepticus increases cell proliferation in the dentate gyrus of adult rats via a 5-HT1A receptor-dependent mechanism. AB - The dentate gyrus continues to produce granule neurons throughout life. Mossy fibers, the axons of granule neurons, undergo atypical sprouting in both clinical and experimental mesial temporal lobe epilepsy. Mossy fiber sprouting (MFS) has been hypothesized to underlie the network reorganization that is thought to produce spontaneously recurring seizures, possibly via the formation of new recurrent excitatory circuits. Hippocampal neurogenesis may be a critical step in the development of MFS, given that it is enhanced by at least 2-fold in the aftermath of pilocarpine-induced status epilepticus. Since it is known that serotonin (5-HT) 1A receptor activation also increases granule cell genesis in the dentate gyrus in rats, and reciprocally, that blockade of this receptor decreases it, we examined whether 5-HT(1A) receptor blockade would prevent the seizure-induced enhancement of neurogenesis. The ability to block seizure-induced neurogenesis would provide a test for its role in the network reorganization, especially in regards to MFS, which might underlie seizure development. In the present study, it was found that blockade of the 5-HT(1A) receptor before and after pilocarpine treatment prevented seizure-induced hippocampal cell proliferation and survival, and, its prevention by chronic treatment with a 5 HT(1A) receptor antagonist (WAY-100,635) did not prevent the development of MFS or spontaneously recurring seizures. Taken together, these results suggest that 5 HT(1A) receptor activation is a critical step in the activation of seizure induced cell proliferation and survival in the dentate gyrus, however, not for the onset of spontaneously recurrent seizures and MFS. PMID- 12646301 TI - Estrogen-mediated effects on depression and memory formation in females. AB - Women are twice as likely to suffer from depression as men. It has been proposed that the ovarian hormones estrogen and progesterone contribute to the higher incidence of this potentially debilitating disorder. Depression can also be accompanied by a loss of cognitive performance. Here we review estrogen-mediated effects on depression and memory formation in females. We propose that changes in levels of estrogen are associated with sex differences in learning as well as changes in affect prior to menses, immediately after pregnancy and during perimenopause and the menopausal transition. Finally, we discuss the animal model of depression known as 'learned helplessness' and describe research from our laboratory demonstrating that exposure to an acute stressful experience compromises a female's later ability to acquire certain types of new memories. This response to stressful experience is opposite to that observed in males and is dependent on the presence of estrogen, and more specifically-changing levels of estrogen. This observation indicates that females and males can use different hormonal and neural mechanisms to respond to the same emotional event and underscore the importance of studying the unique and changing biology of females, especially when considering treatment strategies for depression and stress related illness. PMID- 12646303 TI - Effect of indomethacin on the c-fos expression in AVP and TH neurons in rat brain induced by lipopolysaccharide. AB - The aim of the present study was to investigate the effect of indomethacin on the Fos expression in arginine vasopressin (AVP)-containing neurons in the hypothalamus and tyrosine hydroxylase (TH)-containing neurons in the locus coeruleus (LC) using dual-labeled immunohistochemistry. In the hypothalamus, intraperitoneal (i.p) injection of different doses [2.5 microg/100 g, 125 microg/100 g body weight (b.w.)] of lipopolysaccharide (LPS) induced a significant Fos expression in AVP neurons in the supraoptic nucleus (SON), the magnocellular division (mPVN) and the parvocellular division (pPVN) of the paraventricular nucleus (PVN). Pretreatment with the cyclooxygenase inhibitor indomethacin (0.8 mg/100 g b.w.) significantly blocked the Fos expression in these AVP neurons induced by a low dose of LPS (2.5 microg/100 g) but had no effect on the Fos expression induced by a high dose of LPS (125 microg/100 g). Similarly, in the brain stem, a large number of TH-positive neurons in the LC expressed Fos after administration of either dose of LPS. Indomethacin prevented the Fos expression induced only by a low dose of LPS, but not by a high dose of LPS. These results suggest that the activation of AVP neurons in PVN and SON and TH neurons in LC response to immune challenge might be mediated-at least partially-by prostaglandins. PMID- 12646304 TI - Nitric oxide synthase and arginase expression in the vestibular nucleus and hippocampus following unilateral vestibular deafferentation in the rat. AB - The aim of this study was to investigate the possible relationship between changes in neuronal and endothelial nitric oxide synthase (nNOS and eNOS) and arginase expression in the vestibular nucleus complex and the hippocampus (CA1, CA2/3 and the dentate gyrus (DG) at 10 h or 2 weeks following a unilateral vestibular deafferentation (UVD) in rats. There were no significant differences in nNOS or arginase II expression in the ipsilateral or contralateral VNC at either 10 h or 2 weeks post-UVD. For eNOS, there was only a significant decrease in expression in the ipsilateral VNC at 2 weeks post-UVD (P<0.01). In the hippocampus, the only significant difference in nNOS expression was a decrease in the ipsilateral DG at 2 weeks post-UVD (P<0.05). There was a significant decrease in eNOS expression in the contralateral CA2/3 region at 10 h post-UVD (P<0.01). The only other significant change in eNOS was an increase in the contralateral DG at 10 h post-UVD (P<0.01). Although arginase II was expressed in all regions of the hippocampus, there were no significant differences in arginase II expression at any time point following UVD. These results suggest that the changes in NOS expression that occur in the VNC and hippocampus following UVD are not correlated with one another or with changes in arginase II. PMID- 12646305 TI - Permanent cerebral ischemia induces sustained procaspase 9L increase not controlled by Bcl-2. AB - We have investigated how transgenic overexpression of human Bcl-2 (Hu-Bcl-2) modifies cell death proteins activation in the long-term in a model of permanent cerebral ischemia induced by middle cerebral artery occlusion. Hu-Bcl-2, cytochrome c, caspases 9 and 3 expression were examined by immunoblotting and immunohistochemistry. In wild type mice, 1 day after middle cerebral artery occlusion, cytochrome c released from the mitochondria was detected. Middle cerebral artery occlusion induces a lasting activation of caspases in WT mice from day 3 post-injury. Increased level of caspase 3 is accompanied by a decrease in procaspase 3. In contrast, middle cerebral artery occlusion induced a sustained increase of procaspase 9L and a decrease in procaspase 9S concomitant to caspase 9 production. These events were observed in the operated but not in the unoperated hemisphere. Bcl-2 overexpression blocks cytochrome c release and delays caspases activation. Consequently procaspase 3 decrease was no more observed. However, Bcl-2 overexpression did not influence the middle cerebral artery occlusion-induced changes in procaspases 9 L and S. Fourteen days after middle cerebral artery occlusion the apoptotic cascade was no longer blocked in transgenic mice. Caspases 9 and 3 were increased, procaspase 3 was decreased but procaspase 9L and procaspase 9S remained increased and decreased respectively. Hu Bcl-2 overexpression delays the activation of the cell death molecular machinery but does not control the ischemia-induced change in procaspase 9 L and S. Procaspase 9L increase is a potentially harmful event threatening cells of a rapid destruction when anti-apoptotic treatments by Bcl-2, or caspases inhibitors, are overrun. PMID- 12646306 TI - Mouse Flamingo1/Celsr2 relates neuronal reorganization of the hypertrophic dentate granule cells after kainate injection. AB - Single injection of kainate into the dorsal hippocampus of adult mice induced long-lasting hypertrophy and dispersion of dentate granule cells with dendritic hypertrophy and mossy fiber sprouting that resembled human hippocampal sclerosis. Our previous study indicated that brain derived neurotrophic factor was related to the initiation of these morphological changes. In this study, gene expression of the enlarged hippocampus was examined by differential display to find the gene relating to the progression of the pathological changes. Several genes were identified that were overexpressed in the enlarged dentate gyrus. One of them was highly homologous with mouse Flamingo1/Celsr2, suggesting that mouse Flamingo1/Celsr2 is related to the development of hippocampal sclerosis. PMID- 12646307 TI - Prostaglandin E2 attenuates SR141716A-precipitated withdrawal in tetrahydrocannabinol-dependent mice. AB - The present study aimed to clarify the role of the arachidonic acid cascade in mediating the expression of withdrawal signs in cannabinoid-dependent mice. Mice were injected with Delta(8)-tetrahydrocannabinol (THC) at 20 mg/kg (i.p.) every 12 h, 11 times. When SR141716A, a specific cannabinoid CB1 receptor antagonist, at 10 mg/kg (i.p.) was given 4 h after the last THC injection, withdrawal signs such as forepaw licking, facial preening, grooming, forepaw tremor, head shakes and weight loss were clearly observed. PGE(2) at 0.1, 1.0 and 3.2 microg (per animal; i.c.v.) given prior to SR141716A (10 mg/kg, i.p.) dose-dependently decreased the number of forepaw licking, facial preening, grooming and forepaw tremor episodes. Instead of SR141716A, a cyclooxygenase inhibitor diclofenac at 10 mg/kg (i.p.) also precipitated these withdrawal signs. The results suggest that the expression of THC withdrawal is due to a decrease in prostaglandin levels through inactivation of the arachidonic acid cascade in the brain. PMID- 12646308 TI - NK1 receptors in the medial hypothalamus potentiate defensive rage behavior elicited from the midbrain periaqueductal gray of the cat. AB - Defensive rage in the cat occurs naturally in response to a threat and is also elicited by electrical or chemical stimulation over the rostro-caudal extent of the medial hypothalamus and dorsolateral aspect of the periaqueductal gray (PAG). This behavior is mediated over a descending projection from the hypothalamus to the midbrain PAG. The underlying hypothesis for the present study was that medial hypothalamic defensive rage neurons are excited in two ways: by NK(1) receptors and by an ascending input from the PAG. The first aspect of this hypothesis was tested by eliciting defensive rage by electrical stimulation of the PAG and then microinjecting a selective NK(1) agonist and antagonist into the hypothalamus. Microinjections of 16 or 12 nmol/0.25 microl of the NK(1) agonist, GR 73632, resulted in facilitation of defensive rage. These facilitatory effects were then blocked by pretreatment with the NK(1) antagonist, GR 82334. However, microinjections of GR 82334 alone had no effect. The second aspect of the hypothesis was tested by stimulating defensive rage sites in the PAG and using immunohistochemical methods to test for the presence of c-Fos in the hypothalamus. The results revealed the presence of c-Fos immunoreactivity in the medial but not lateral hypothalamus. Overall, the findings indicate that NK(1) receptors in the medial hypothalamus facilitate defensive rage elicited from PAG neurons whose axons project back to the medial hypothalamus. The likely ethological significance of the ascending input is that it allows for potentiation and prolongation of defensive rage in response to a threatening stimulus. PMID- 12646310 TI - Feeding induced by microinjections of NMDA and AMPA-kainate receptor antagonists into ventral striatal and ventral pallidal areas of the pigeon. AB - The participation of glutamatergic circuits of the ventral basal ganglia in feeding-related regulatory mechanisms has been extensively indicated in primate and rodent species. In avian species, it has been shown that ICV injections of MK 801 or of CNQX increase food intake and reduce the latency of feeding initiation in free-feeding pigeons. In the present study, the effects of local injections of MK-801 (6 nmol), CNQX (160 nmol) or vehicle (0.2 microl) into a number of ventral striatopallidal nuclei on feeding, drinking and non-ingestive (sleep, preening) spontaneous behaviors were investigated in free-feeding pigeons (Columba livia). Intense feeding responses associated with an increased duration of feeding behavior were consistently recorded after injections of MK-801 or CNQX into the medial two-thirds of the tuberculum olfactorium (TO), the ventral aspect of lobus parolfactorium (LPOv), or the ventral pallidum (VP). In contrast, the latency of feeding initiation was unaffected by these treatments. No changes in drinking, preening or sleep responses were observed after drug injections into the TO/LPOv/VP area. These data indicate that glutamate-mediated circuits in the TO/LPOv/VP area can play an inhibitory role in feeding behavior in this species, contributing to the conclusion of a feeding bout, thus delaying satiation processes, and that these effects may be mediated by AMPA and NMDA receptors. Additionally, our data support the notion that a region functionally and anatomically comparable to the mammalian accumbens shell may be present in the TO/LPOv/VP region of the pigeon, and that the existence of a glutamatergic circuit in the ventral striatum controlling feeding-related phenomena may represent a highly conserved attribute throughout the amniote's evolution. PMID- 12646309 TI - Effect of 17 beta-estradiol on gene expression in lumbar spinal cord following sciatic nerve crush injury in ovariectomized mice. AB - Previously, we observed that estrogen treatment enhances regeneration of the sciatic nerve after crush injury [Brain Res. 943 (2002) 283]. In this research, we studied expression of estrogen receptors and effects of estrogen on gene expression in the lumbar spinal cord, following sciatic nerve crush injury. Using the Atlas Mouse 1.2 Array, changes in the expression of 267 of 1176 genes were registered 4 days after nerve injury. Those genes that exhibited a change in signal intensity ratios of 2-fold or greater were selected as up-regulated (42) or down-regulated (21). In estrogen treated mice, we have observed up-regulation of the genes known to control apoptosis, cell proliferation, and growth, which might account for the positive effects of estrogen on the regeneration of motor neurons. Immunohistochemical staining revealed estrogen receptor-alpha and estrogen receptor-beta localized in the nucleus and cytoplasm of lumbar motor neurons, and in the regenerating neurites of the sciatic nerve. Expression of estrogen receptor-alpha and estrogen receptor-beta mRNA in lumbar spinal cord was shown by traditional RT-PCR. Using real-time quantitative RT-PCR, we demonstrated increased expression of estrogen receptors-alpha and -beta mRNA on the injured side of the lumbar spinal cord. Western blot analysis showed the accumulation of ERs in regenerating sciatic nerve, and revealed a 40% increase of activated ERK1/2 in estrogen treated mice, compared to placebo. Our findings indicate that: (i). axotomized motor neurons increase expression of estrogen receptors-alpha and -beta mRNA, (ii). estrogen mediates the expression of genes which accelerate the growth and maturation of axons, and (iii). estrogen receptors are transported from the perikaryon into regenerating neurites, and estrogen promotes regeneration locally through the non-genomic ERK-activated signaling pathway. PMID- 12646311 TI - Intrinsic and synaptic properties of the dorsomedial nucleus of the intercollicular complex, an area known to be involved in distance call production in Bengalese finches. AB - The dorsomedial nucleus of the intercollicular complex (DM) of the midbrain in the Bengalese finch is essential for the vocal production of distance calls that have sexually-dimorphic acoustic structures in the adult. Anatomical tracing of the vocal control system shows that DM neurons of adult males receive axonal inputs from the robust nucleus of the archistriatum (RA), and the inputs are considered to be crucial for the male-typical features of distance calls. In order to investigate the neural mechanisms underlying distance call patterns of male finches in DM, we characterized neurons in DM and examined their synaptic responses to RA inputs in brain slice preparations. By using whole-cell recording techniques, we could classify at least three types of neurons based on electrophysiological and morphological characteristics. Type I neurons exhibited regular and high-frequency trains of action potentials in response to depolarizing current pulses. Type II neurons had large somata and action potential trains accommodating during depolarization. Type III neurons were characterized by a few spikes followed by a slow depolarization during current injection. Their somata were markedly small and their axons often projected toward the contralateral DM or the thalamic nucleus uvaeformis (Uva). In all these cell types, electrical stimulation of an area including DM-projecting RA axons often elicited both excitatory postsynaptic currents (EPSCs) mediated mainly by non-NMDA glutamate receptors and inhibitory postsynaptic currents (IPSCs) mediated by GABA(A) receptors. These intrinsic properties of DM neurons and their excitatory and inhibitory synaptic inputs may play important roles in generating the acoustic patterns of distance calls in male finches. PMID- 12646312 TI - Melatonin inhibition of nicotine-stimulated dopamine release in PC12 cells. AB - Melatonin, a pineal hormone, modifies numerous physiologic processes including circadian rhythms and sleep. In specific tissues, melatonin appears to have an inverse relationship with dopamine. To examine this relationship, a pheochromocytoma cell line (PC12) was used to determine the extent of melatonin's ability to inhibit nicotine-stimulated dopamine release. Multiple experiments were conducted that examined: (1). the dose response of acute melatonin (5 min); (2). the effects of chronic melatonin (16 h pre-exposure); (3). the effects of prior nicotine or melatonin exposure (5 min) on melatonin's ability to alter dopamine release from a second 5-min nicotine exposure; and (4). the role of melatonin receptors (by pertussis toxin inhibition) on nicotine-stimulated dopamine release. In the dose response studies, melatonin inhibited nicotine stimulated dopamine release with an ED50 of 8.6 microM. Chronic exposure to melatonin had no effect on melatonin's acute inhibition of nicotine-stimulated dopamine release. Prior nicotine or melatonin exposure had little effect on subsequent melatonin or nicotine exposure, except that the cells exposed to nicotine were not responsive to a second exposure to nicotine. Blockade of melatonin receptor function by pre-exposure to pertussis toxin (16 h) did not prevent melatonin's inhibition of nicotine-stimulated dopamine release. However, the toxin-treated cells were less inhibited by melatonin when compared to control cells suggesting a partial role for melatonin receptors. These results indicate that melatonin can acutely inhibit nicotine-stimulated dopamine release in PC12 cells. This model system allows detailed examination of melatonin's cellular actions as well as supporting a role for melatonin on neuronal dopamine release. PMID- 12646313 TI - Involvement of the raphe pallidus in the suppressive effect of preoptic warming on non-shivering thermogenesis in rats. AB - Thermogenesis in the brown adipose tissue (BAT) is activated by the stimulation of the ventromedial hypothalamic nucleus (VMH). Local warming of the preoptic area (PO) suppresses this response. Injection of the GABA(A) receptor antagonist bicuculline into the caudal periaqueductal gray (cPAG), where excitatory neurons for BAT thermogenesis are located, did not influence the suppressive effect of PO warming. On the other hand, after bicuculline injection into the raphe pallidus, where excitatory neurons for BAT thermogenesis are also located, VMH stimulation produced BAT thermogenesis even during PO warming. The present results suggest that the inhibitory signal from the PO reaches the raphe pallidus and not the cPAG for the control of BAT thermogenesis. PMID- 12646314 TI - Serotonin phase-shifts the mouse suprachiasmatic circadian clock in vitro. AB - The mammalian circadian clock in the suprachiasmatic nucleus (SCN) receives multiple afferent signals that could potentially modulate its phase. One input, the serotonin (5-HT) projection from the raphe nuclei, has been extensively investigated in rats and hamsters, yet its role(s) in modulating circadian clock phase remains controversial. To expand our investigation of 5-HT modulation of the SCN clock, we investigated the phase-shifting effects of 5-HT and its agonist, (+)8-hydroxy-2-(di-n-propylamino)tetralin (DPAT), when applied to mouse SCN brain slices. 5-HT induced 2-3 h phase advances when applied during subjective day, while non-significant phase shifts were seen after 5-HT application at other times. These phase shifts were completely blocked by the 5 HT antagonist, metergoline. DPAT also induced phase shifts when applied during mid-subjective day, and this effect appeared dose-dependent. Together, these results demonstrate that the mouse SCN, like that of the rat, is directly sensitive to in vitro phase-resetting by 5-HT. PMID- 12646315 TI - [1-13C]Glucose entry in neuronal and astrocytic intermediary metabolism of aged rats. A study of the effects of nicergoline treatment by 13C NMR spectroscopy. AB - Age-related changes in glucose utilization through the TCA cycle were studied using [1-13C]glucose and 13C, 1H NMR spectroscopy on rat brain extracts. Significant increases in lactate levels, as well as in creatine/phosphocreatine ratios (Cr/PCr), and a decrease in N-acetyl-aspartate (NAA) and aspartate levels were observed in aged rat brains as compared to adult animals following glucose administration. The total amount of 13C from [1-13C]glucose incorporated in glutamate, glutamine, aspartate and GABA was significantly decreased in control aged rat brains as compared to adult brains. The results showed a decrease in oxidative glucose utilization of control aged rat brains. The long-term nicergoline treatment increased NAA and glutamate levels, and decreased the lactate levels as well as the Cr/PCr ratios in aged rat brains as compared to adult rats. The total amount of 13C incorporated in glutamate, glutamine, aspartate, NAA and GABA was increased by nicergoline treatment, showing an improvement in oxidative glucose metabolism in aged brains. A significant increase in pyruvate carboxylase/pyruvate dehydrogenase activity (PC/PDH) in the synthesis of glutamate in nicergoline-treated aged rats is consistent with an increase in the transport of glutamine from glia to neurons for conversion into glutamate. In adult rat brains, no effect of nicergoline on glutamate PC/PDH activity was observed, although an increase in PC/PDH activity in glutamine was, suggesting that nicergoline affects the glutamate/glutamine cycle between neurons and glia in different ways depending on the age of animals. These results provide new insights into the effects of nicergoline on the CNS. PMID- 12646316 TI - Central projections of sensory innervation of the rat superficial temporal artery. AB - Elucidating the central sensory projection pathways of extra- and intracranial vessels appears to be of fundamental importance for understanding the pathogenetic mechanisms of primary headaches. In this paper, two kinds of tracers, choleragenoid (cholera toxin subunit b, CTb) and wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP), were used to transganglionically label the central sensory projections of the innervation of the superficial temporal artery (STA). Following either of the tracers applied on the adventitia of the STA, labelled terminations were found mainly in the ipsilateral C1-C3 spinal dorsal horns. Sparse labelling was also found in the interpolar and caudal parts of the spinal trigeminal nucleus. In the spinal cord, CTb labelled profiles were mainly located in laminae III and IV, whereas WGA-HRP labelled profiles were mainly located in laminae I and II. In the medulla, CTb but not WGA-HRP labelled terminals were found in a small dorsolateral extension of the cuneate nucleus. The present results indicate that the primary sensory nervous center of the STA is located in the rostral cervical spinal dorsal horn. The caudal parts of the spinal trigeminal nucleus, which has been demonstrated as a center of pain and temperature sensations of the head and face, transmits limited information from the STA to higher nervous centers. PMID- 12646317 TI - Distribution of Fos labeling in the inferior olive following transient blockade of the VIIIth cranial nerve. AB - The sodium channel blocker, tetrodotoxin (TTX), is an effective tool for blockade of action potentials in neurons. Unilateral transtympanic administration of 3 mM TTX produced behavioral symptoms paralleling those previously reported following unilateral vestibular ablation. Behavioral symptoms were evident as early as 15 min post-TTX. Fos immunocytochemistry revealed an initial bilateral distribution of Fos in the inferior olive (IO) followed by an almost exclusively unilateral distribution of Fos. By 1 h, Fos was predominantly localized in subdivisions of the IO contralateral to TTX treatment. Fos labeling in the IO was most pronounced at 2- and 6-h survival times and was localized in the contralateral IOA, IOB, IOC, IOBe, and IOK subdivisions and bilaterally in the IOM and IODM. Other regions of the brainstem including the vestibular nuclei, prepositus hypoglossi, dorsal paragigantocellular reticular nucleus, nucleus of the tractus solitarius and locus coeruleus also exhibited altered patterns of Fos labeling following TTX. The finding that Fos activity in the IO is initially bilateral and then rapidly becomes unilateral has not been reported for the traditional vestibular ablation models and may be unique to the TTX model. In addition, since altered Fos activity is readily detected in the IO at time-points prior to detectable changes in Fos in the central vestibular complex it is possible that the IO is particularly sensitive to events precipitated by unilateral vestibular disturbance. PMID- 12646318 TI - Cholinergic deficits in the septal-hippocampal pathway of the SAM-P/8 senescence accelerated mouse. AB - Senescence accelerated prone mouse strains (SAM-P) and resistant strains (SAM-R) have proven useful in elucidating aspects of the aging process. The senescence accelerated mouse SAM-P/8 strain exhibits severe age-related learning and memory impairments well before the median age of survival. Disruption of the brain cholinergic system produces learning and memory impairments as severe as those seen in aging SAM-P/8 mice. Therefore, we compared the effects of aging on cholinergic parameters in the septal-hippocampal pathway, a region known to play a role in learning and memory, in SAM-P/8 mice and mice of the senescence resistant SAM-R/1 strain. Between 4 and 12 months of age we observed a 40-50% decrease in choline acetyltransferase (ChAT) activity in two of three subregions of the hippocampus in the SAM-P/8, but not the SAM-R/1 strain. Between 4 and 12 months, SAM-P/8 mice also showed a 40-50% decrease in ChAT activity in the septal region that was maximal by 8 months of age. By contrast, these age-related changes were not observed in the control SAM-R/1 mouse strain. The changes in ChAT in the SAMP/8 mouse strain were limited to the septal-hippocampal cholinergic pathway. There were no differences in ChAT activity in the nucleus basalis of Meynert, nor any of several neocortical areas to which it projects. To determine the neurochemical specificity of these alterations, the activity of glutamic acid decarboxylase (GAD), was also measured in the septum and hippocampus of SAM-P/8 mice. There were no age-related alterations in the hippocampus, but a significant 50% increase in GAD activity in the septal nucleus at 12 months of age. There were no age-related alterations in either nicotinic (3H-cytisine) or muscarinic (3H-QNB) cholinergic receptor binding in the cortex or hippocampus of SAM-P/8 mice. However, there were significant strain differences. At 2 months of age, 3H-QNB binding was higher in hippocampus of the SAM-R/1 than in SAM-P/8 mice. Similarly, 3H-cytisine binding in cortex of SAM-R/1 mice was higher at both 2 and 13 months than in SAM-P/8 mice. The results suggest that a compromised septal-hippocampal cholinergic pathway may contribute to the previously reported early onset of impaired learning and memory in the SAM-P/8 mouse strain. PMID- 12646319 TI - Nimodipine potentiates light-induced phase shifts of circadian activity rhythms but not c-fos expression in the suprachiasmatic nucleus of mice. AB - The present study assessed whether treatment with the L-type calcium channel antagonist nimodipine affects the responsiveness of the circadian pacemaker to light in C3H/HeN mice. Nimodipine (10 mg/kg, sc) increased the magnitude of light induced phase delays (P<0.01) and c-fos mRNA expression in the paraventricular nuclei (P<0.01), but not in the suprachiasmatic nuclei (SCN). These results suggest that nimodipine may affect phase shifts of circadian activity rhythms through a mechanism independent of c-fos expression in the SCN. PMID- 12646320 TI - The expression and localization of heme oxygenase in the adult guinea pig cochlea. AB - Reverse transcription-polymerase chain reaction demonstrated the expression of heme oxygenase-1 and heme oxygenase-2 (HO-2) mRNA in the cochlea, but the former was considered to be much lower than the latter. The HO-2-like immunoreactivity was observed mainly in supporting cells in the organ of Corti and in the spiral ganglion cells. However, in the vascular walls of the cochlea little immunoreactivity was detected. These results suggest that HO-2 is involved in the auditory neurotransmission and micromechanics of the organ of Corti. PMID- 12646321 TI - Genetic engineering of a zeaxanthin-rich potato by antisense inactivation and co suppression of carotenoid epoxidation. AB - Zeaxanthin is an important dietary carotenoid but its abundance in our food is low. In order to provide a better supply of zeaxanthin in a staple crop, two different potato (Solanum tuberosum L.) varieties were genetically modified. By transformation with sense and antisense constructs encoding zeaxanthin epoxidase, zeaxanthin conversion to violaxanthin was inhibited. Both approaches (antisense and co-suppression) yielded potato tubers with higher levels of zeaxanthin. Depending on the transgenic lines and tuber development, zeaxanthin content was elevated 4 to 130-fold reaching values up to 40 microg/g dry weight. As a consequence of the genetic manipulation, the amount of violaxanthin was diminished dramatically and in some cases the monoepoxy intermediate antheraxanthin accumulated. Between one and eight copies of the sense or antisense epoxidase gene fragments were integrated into the genome. In addition, most of the transformants with higher zeaxanthin levels showed also increased total carotenoid contents (up to 5.7-fold) and some of them exhibited reduced amounts of lutein. The increase in total carotenoids suggests that the genetic modification affects the regulation of the whole carotenoid biosynthetic pathway in potato tubers. Northern blot analysis demonstrated that upregulation of carotenogenesis in the transgenics is accompanied by substantial higher phytoene synthase transcript levels in 6-week-old tubers and a very slight increase of the beta-carotene hydroxylase transcript. The amount of the deoxyxylulose 5-phosphate synthase mRNA was very similar in wild type and transformed tubers. Abscisic acid content of tubers remained unchanged whereas alpha-tocopherol was 2 to 3 fold elevated in the transformants. PMID- 12646323 TI - Computing positional isotopomer distributions from tandem mass spectrometric data. AB - The isotopomer distributions of metabolites are invaluable pieces of information in the computation of the flux distribution in a metabolic network. We describe the use of tandem mass spectrometry with the daughter ion scanning technique in the discovery of positional isotopomer distributions (PID). This technique increases the possibilities of mass spectrometry since given the same fragment ions, it uncovers more information than the full scanning mode. The mathematics of the new technique is slightly more complicated than the techniques needed by full scanning mode methods. Our experiments, however, show that in practice the inadequacy of the fragmentation of amino acids in the tandem mass spectrometer does not allow uncovering the PID exactly even if the daughter ion scanning is used. The computational techniques have been implemented in a MATLAB application called PIDC (Positional Isotopomer Distribution Calculator). PMID- 12646322 TI - Mathematical modeling of in vitro enzymatic production of 2-Keto-L-gulonic acid using NAD(H) or NADP(H) as cofactors. AB - A 2-Keto-L-gulonic acid (2-KLG) production process using stationary Pantoea citrea cells and a Corynebacterium 2,5-diketo-D-gluconic acid (2,5-DKG) reductase enzyme has been developed which may represent an improved method of vitamin C biosynthesis. Experimental data was collected using the F22Y/A272G 2,5-DKG reductase mutant and NADP(H) as a cofactor. An extensive kinetic analysis was performed and a kinetic rate equation model for this process was developed. A recent protein engineering effort has resulted in several 2,5-DKG reductase mutants exhibiting improved activity with NADH as a cofactor. The use of NAD(H) in the bioreactor may be preferable due to its increased stability and lower cost. The kinetic parameters in the rate equation model have been replaced in order to predict 2-KLG production with NAD(H) as a cofactor. The model was also extended to predict 2-KLG production in the presence of a range of combined cofactor concentrations. This analysis suggests that the use of the F22Y/K232G/R238H/A272G 2,5-DKG reductase mutant with NAD(H) combined with a small amount of NADP(H) could provide a significant cost benefit for in vitro enzymatic 2-KLG production. PMID- 12646324 TI - Changes of pentose phosphate pathway flux in vivo in Corynebacterium glutamicum during leucine-limited batch cultivation as determined from intracellular metabolite concentration measurements. AB - Corynebacterium glutamicum is an important organism for the industrial production of amino acids such as lysine. In the present study time-dependent changes in the oxidative pentose phosphate pathway activity, an important site of NADPH regeneration in C. glutamicum, are investigated, whereby intracellular metabolite concentrations and specific enzyme activities in two isogenic leucine auxotrophic strains differing only in the regulation of their aspartate kinases were compared. After leucine limitation only the strain with a feedback-resistant aspartate kinase began to excrete lysine into the culture medium. Concomitantly, the intracellular NADPH to NADP concentration ratio increased from 2 to 4 in the non-producing strain, whereas it remained constant at about 1.2 in the lysine producing strain. From these data the in'vivo flux through the pentose phosphate pathway was calculated. These results were used to approximate the total NADPH regeneration by glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and isocitrate dehydrogenase, which agreed fairly well with the calculated demands for biomass formation and lysine biosynthesis. The analysis allowed to conclude that NADPH regeneration in the pentose phosphate pathway is essential for lysine biosynthesis in C. glutamicum. PMID- 12646325 TI - Research to practice: adoption of naltrexone in alcoholism treatment. AB - Naltrexone, a prescription medication, was approved in December 1994 as an adjunct to counseling in treatment of alcoholism and alcohol abuse, representing the first new medication for alcoholism in several decades. Initial controlled trials indicated that it is effective in preventing relapse, while later trials show mixed results. Although many physicians and others treating alcoholism have found naltrexone to be very helpful in treatment, it is still a technology that has not been used widely. In this study, we examine which clinicians have adopted naltrexone into practice for what reasons, and what clinical and nonclinical factors acted as barriers to its use. In our mail survey of alcoholism treatment clinicians, 80% of physicians and 45% of nonphysicians report prescribing or recommending naltrexone at least rarely, but only 15% of physicians, even among addiction specialists, prescribe naltrexone often. The strongest barriers to adoption of naltrexone were financing and inadequate knowledge about the medication, followed by lack of sufficient evidence regarding effectiveness. Clinicians were most likely to adopt naltrexone if they were affiliated with treatment programs that actively promoted its use. We conclude that in order for a new substance abuse treatment medication to be widely adopted in clinical practice, information about it must be properly directed, clinicians must be convinced of its effectiveness, it must be adequately financed, and the treatment organizations in which clinicians work must promote its use. PMID- 12646327 TI - The relationship between attitudes toward pregnancy and contraception use among drug users. AB - Use of alcohol and drugs during pregnancy is regarded as a serious problem in the medical field. The current study explored gender and ethnic differences in attitudes and behaviors related to sexuality and contraceptive use among European American, Mexican American and Native American female and male clients in drug recovery programs. The study also examined the relationship between sexual abuse, concern for pregnancy, concern for contracting sexually transmitted diseases and contraception use among women drug users. Results showed gender and ethnic differences in attitudes toward pregnancy, abortion and contraceptive use. In addition, both Mexican American and Native American women who had a high concern for pregnancy were more likely to use birth control when they reported sexual abuse than when they reported no abuse. In contrast, European American women with a high concern for pregnancy showed a tendency to use birth control less when they reported abuse than when they did not. The observed attitudinal and behavioral differences between genders and ethnic groups are discussed in relation to design of treatment program components to increase contraceptive use and decrease unwanted pregnancies among drug users. PMID- 12646326 TI - Prolonged nicotine patch use in quitters with past abstinence-induced depressed mood. AB - To test the efficacy of sustained nicotine patch use among at-risk smokers, 55 smokers with a history of abstinence-induced depressed mood were randomly assigned to either Nicotine Maintenance or Standard Treatment following preliminary high-dose patch treatment. The Nicotine Maintenance group received 21 mg transdermal nicotine for 8 additional weeks; the Standard Treatment group followed a tapered dosing regimen. Significant differences favoring the Nicotine Maintenance group were found in self-reported craving but not withdrawal. No difference was observed in continuous abstinence or in relapse rates. When dropouts who did not relapse during patch use were classified as successful, however, the Nicotine Maintenance group had significantly lower relapse rates. Rate of lapse in the Nicotine Maintenance group during post-trial tapering did not differ significantly from that in the Standard Treatment group during tapering in the trial, suggesting that the benefits of sustained dosing may persist only as long as dosing continues. PMID- 12646328 TI - Rationales, obstacles and strategies for local outcome monitoring systems in substance abuse treatment settings. AB - The need for outcome monitoring systems (OMS) in substance abuse treatment is now such a pervasive position that it has achieved 'motherhood' stature. Yet, for a variety of reasons, implementation of systematic, ongoing monitoring continues to lag in most settings. The position taken in this article is that a local approach to monitoring is a viable alternative to large scale monitoring programs when such programs are unavailable. Moreover, local OMS can supplement these more generic programs by providing data that are more in tune with the orientation and program development of a specific setting. This article argues that local OMS can be developed with a reorganization of customary documentation procedures, a modest investment of personnel and specialized technical resources, a shorter term view to outcome, and a commitment to evidence-based treatment. PMID- 12646329 TI - Outcomes at 1 and 5 years for older patients with alcohol use disorders. AB - Older patients with alcohol use disorders who had gone through residential treatment were compared with matched groups of young and middle-aged patients (N = 432 in each age group) on their 1- and 5-year outcomes, use of continuing care services, and outcome predictors. Older patients had better outcomes than did young and middle-aged patients but had comparable levels of continuing substance abuse care and 12-step self-help group involvement. Similar factors predicted outcomes across the age groups. Longer duration of continuing substance abuse care and greater self-help group involvement were related to better outcomes, as were patients' attitudes and coping strategies at program discharge. The findings indicate that older patients with alcohol use disorders respond to age-integrated substance abuse treatment programs at least as well as do younger patients and are equally involved in formal and informal continuing substance abuse care. PMID- 12646330 TI - A national study of the substance abuse treatment workforce. AB - This study's purpose is to gain a current perspective on the substance abuse treatment field's workforce. The data are from the Retrospective Study of treatment professionals designed to document how the Treatment Improvement Protocols published by the Center for Substance Abuse Treatment have influenced the implementation of best practices. The Retrospective Study consisted of a two wave cross-sectional survey with telephone follow-up. Data for this study were from demographic information on Wave 1 study participants, which had a response rate of 80.1% (N = 3,267). The results of the study showed that most treatment professionals are White (84.5%) and middle-aged (i.e., between 40 and 55 years old) and slightly more are female (50.5.0%) than male (49.5%). Treatment professionals tend to enter the field and stay in it, and almost 80.0% of respondents possess a bachelor's degree or higher. In addition, most treatment professionals are licensed or certified and treat clients from different racial and ethnic backgrounds than themselves. Implications for the provision of treatment services are discussed. PMID- 12646331 TI - Survey of smoking cessation services in Canadian addiction programs. AB - Many alcohol and drug treatment programs now offer smoking cessation (SC) services to their clients. Little is known about the prevalence of such services in Canadian addiction programs. A telephone survey was conducted of all Canadian adult substance abuse programs with outpatient, day program, and residential treatment to determine: (a) whether the program offered help in quitting smoking; (b) the type of SC services; (c) the overall priority placed on quitting smoking; and (d) for residential programs, what type of smoking ban (indoors, outdoors, or both), if any, is imposed. Over half (54%) of the 223 programs that responded to the survey reported providing help in quitting smoking. Twenty-three programs had formal SC services, most often in the form of group treatment. The majority of programs surveyed stated their program placed 'very little' emphasis on smoking. Smoking was relatively unrestricted in residential programs. The implications of these findings for substance abuse treatment are discussed. PMID- 12646332 TI - Feasibility of referring drug users from a needle exchange program into an addiction treatment program: experience with a mobile treatment van and LAAM maintenance. AB - We evaluated program entry, retention, and early treatment response of needle exchange program (NEP) attenders referred to a drug treatment program using levomethadyl acetate hydrochloride (LAAM). Of 163 referrals, 114 (70%) entered the program, and 84% were retained for at least 90 days. Comparing baseline and follow-up visits after 1 month, there were significant reductions in the Addiction Severity Index subscale scores for drug and alcohol use and legal situation. We observed a 31% and 22% reduction in heroin- and cocaine-positive urine tests, respectively (p < .0001). Although LAAM is no longer considered a first line treatment for heroin addiction, these results demonstrate the feasibility of utilizing long-acting agonist therapies such as LAAM to treat opioid dependence among NEP attenders. PMID- 12646333 TI - Cognitive disparity in schizophrenics with and without cocaine dependency. AB - Although cognition has been investigated in individuals with schizophrenia and in non-schizophrenic cocaine abusers, few studies have focused on cocaine-abusing schizophrenics. Previous studies have shown contradictory results despite the fact that individuals with schizophrenia and cocaine dependence have worse long term outcomes, and that each disorder separately is associated with neuropsychological impairment. The present study intended to clarify these inconsistencies with a comprehensive neuropsychological battery. Twenty-four cocaine-dependent schizophrenics and 23 non-drug abusing schizophrenics were recruited from the VA. Participants were administered tests focusing on motor skills, processing speed, attention, concentration, and executive functioning. While individuals with schizophrenia and cocaine dependence performed worse on the Grooved Peg Board and the Stroop A, the non-drug abusing schizophrenics performed worse on Trails Part A and B. However, a MANOVA failed to show group differences in overall neuropsychological performance. These findings are similar to the existing literature and suggest that cocaine may compromise motor functioning. PMID- 12646334 TI - Trends in comprehensive service availability in outpatient drug abuse treatment. AB - Comprehensive medical and psychosocial services are essential to quality addiction treatment, but their availability declined in the 1980s. To determine whether this downward trend in the availability of comprehensive services continued in the 1990s, we analyzed data from a national panel study of outpatient substance abuse treatment units in 1990, 1995, and 2000. Response rates were greater than 85%. Regarding the availability of comprehensive services, including physical examinations, routine medical care, mental health services, financial counseling and employment counseling, administrators reported whether any substance abuse treatment client received the service in the past year. With the exception of physical examinations, whose reported availability increased from 1990 to 1995, and financial counseling, whose reported availability decreased during the same time, the reported availability of comprehensive services changed little during the 1990s. These findings highlight the continuing need to monitor access to comprehensive services and other quality markers in addiction treatment over time. PMID- 12646335 TI - Introduction: rosuvastatin--an efficacy assessment based on pooled trial data. PMID- 12646336 TI - Efficacy of rosuvastatin compared with other statins at selected starting doses in hypercholesterolemic patients and in special population groups. AB - A total of 5 randomized, double-blind trials in patients with hypercholesterolemia were prospectively designed to allow pooling of plasma lipid data after 12 weeks of treatment. The purpose was (1) to compare rosuvastatin 5 and 10 mg with atorvastatin 10 mg (data from 3 of the 5 trials); (2) to compare rosuvastatin 5 and 10 mg with simvastatin 20 mg and pravastatin 20 mg (data from 2 of the 5 trials); and (3) to summarize overall efficacy and subset analyses of rosuvastatin data from all 5 trials. Rosuvastatin 5 mg (n = 390) and 10 mg (n = 389) reduced low-density lipoprotein (LDL) cholesterol significantly more than did atorvastatin 10 mg (n = 393) (41.9% and 46.7% vs 36.4%, both p <0.001). Treatment with rosuvastatin 5 mg (n = 240) and 10 mg (n = 226) also resulted in significantly greater reductions in LDL cholesterol compared with both simvastatin 20 mg (n = 249) and pravastatin 20 mg (n = 252) (40.6% and 48.1% vs 27.1% and 35.7%, all p <0.001). Significant differences favoring rosuvastatin 10 mg were also observed for total cholesterol, high-density lipoprotein (HDL) cholesterol, non-HDL cholesterol, apolipoprotein (apo) B, and apo A-I versus atorvastatin 10 mg, and for total cholesterol, HDL cholesterol, triglycerides, non-HDL cholesterol, and apo B versus simvastatin 20 mg and pravastatin 20 mg. Analyses of all the rosuvastatin 10 mg data (n = 615) from the 5 trials in subgroups defined by age > or =65 years, female sex, postmenopausal status, hypertension, atherosclerosis, type 2 diabetes, and obesity showed that rosuvastatin had consistent efficacy across patient subgroups. PMID- 12646338 TI - Guidelines for lowering lipids to reduce coronary artery disease risk: a comparison of rosuvastatin with atorvastatin, pravastatin, and simvastatin for achieving lipid-lowering goals. AB - Both the National Cholesterol Education Program (NCEP) Adult Treatment Panel (ATP) III and the Second Joint Task Force of European Societies guidelines have established low-density lipoprotein (LDL) cholesterol goals for lipid-lowering treatment to reduce the risk of coronary artery disease. Data from 3 trials that compared rosuvastatin 10 mg (n = 389) with atorvastatin 10 mg (n = 393) and 2 trials that compared rosuvastatin 10 mg (n = 226) with pravastatin 20 mg (n = 252) and simvastatin 20 mg (n = 249) were pooled separately to compare the achievement of LDL cholesterol goals over 12 weeks of treatment in hypercholesterolemic patients. Noncomparative pooling of rosuvastatin 10 mg results from all 5 trials (n = 615) showed that 80% achieved NCEP ATP III goals and 81% achieved the European goal of <3.0 mmol/L. Compared with atorvastatin 10 mg, significantly more patients treated with rosuvastatin 10 mg achieved their ATP III (76% vs 53%) and European (82% vs 51%) goals (p <0.001). Also, in comparisons with simvastatin 20 mg and pravastatin 20 mg, 86% of patients treated with rosuvastatin 10 mg achieved ATP III goals, compared with 64% of simvastatin treated patients and 49% of pravastatin-treated patients (p <0.001). The proportions of patients who achieved the European goal were 80%, 48%, and 16% for rosuvastatin 10 mg, simvastatin 20 mg, and pravastatin 20 mg, respectively, in this comparison (all p <0.001). A total of 71% of patients treated with rosuvastatin 10 mg who had triglyceride levels > or =200 mg/dL met both their LDL cholesterol and their non-high-density lipoprotein cholesterol goals. PMID- 12646344 TI - John T. Edsall: 3 November 1902-12 June, 2002. PMID- 12646340 TI - Lipid and apolipoprotein ratios: association with coronary artery disease and effects of rosuvastatin compared with atorvastatin, pravastatin, and simvastatin. AB - Plasma lipid and apolipoprotein ratios that include both an atherogenic and an antiatherogenic lipid component (eg, total cholesterol/high-density lipoprotein [HDL] cholesterol ratio, low-density lipoprotein [LDL] cholesterol/HDL cholesterol ratio, non-HDL cholesterol/HDL cholesterol ratio, and apolipoprotein [apo] B/apo A-I ratio) have been found to be strong predictors of coronary artery disease (CAD) risk. Three trials that compared the effects of rosuvastatin 10 mg versus atorvastatin 10 mg and 2 trials that compared the effects of rosuvastatin 10 mg versus simvastatin 20 mg and pravastatin 20 mg on lipid ratios in patients with hypercholesterolemia were prospectively designed for pooled analysis. At 12 weeks, in the 3-trial pooled analysis, rosuvastatin 10 mg (n = 389) showed significantly greater reductions in all 4 lipid ratios compared with atorvastatin 10 mg (n = 393) (p <0.001). The mean percent reduction from baseline in the LDL cholesterol/HDL cholesterol ratio was 51% in patients treated with rosuvastatin 10 mg versus 39% in patients treated with atorvastatin 10 mg. In the 2-trial pooled analysis, treatment with rosuvastatin 10 mg (n = 226) also resulted in significantly greater reductions in all 4 lipid ratios compared with both simvastatin 20 mg (n = 249) and pravastatin 20 mg (n = 252) (p <0.001). Mean percent reductions from baseline in the LDL cholesterol/HDL cholesterol ratio were 52%, 39%, and 30% for rosuvastatin 10 mg, simvastatin 20 mg, and pravastatin 20 mg, respectively, in these 2 trials. PMID- 12646342 TI - Efficacy of rosuvastatin 10 mg in patients with the metabolic syndrome. AB - The constellation of risk factors known as the metabolic syndrome increases the risk of coronary artery disease at any low-density lipoprotein (LDL) cholesterol level. We performed an exploratory analysis of data from 5 trials to study the effects of rosuvastatin 10 mg on lipid levels and ratios in hypercholesterolemic patients (LDL cholesterol > or =160 mg/dL and <250 mg/dL) who met a modified National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) definition of the metabolic syndrome. Of 580 patients completing 12 weeks of treatment with rosuvastatin 10 mg, 194 (33%) met the definition of the metabolic syndrome by exhibiting > or =3 of the following: body mass index >30; triglycerides > or =150 mg/dL; high-density lipoprotein (HDL) cholesterol <40 mg/dL in men and <50 mg/dL in women; blood pressure > or =130/> or =85 mm Hg or receiving current medication for hypertension; and fasting blood glucose > or =110 mg/dL. Patients with the metabolic syndrome had higher triglyceride, non-HDL cholesterol, apolipoprotein B, and lipid ratios, and lower HDL cholesterol and apolipoprotein A-I levels, at baseline compared with patients without the metabolic syndrome. In patients with the metabolic syndrome, rosuvastatin 10 mg improved LDL cholesterol (-47%), non-HDL cholesterol (-43%), non-HDL cholesterol/HDL cholesterol ratio (-47%), apolipoprotein B (-37%), apolipoprotein B/apolipoprotein A-I ratio (-40%), triglycerides (-23%), apolipoprotein A-I (+7%), and HDL cholesterol (+10%)-in a manner similar to that in hypercholesterolemic patients who did not meet these criteria. Among patients who met the metabolic syndrome criteria and who had triglycerides > or =200 mg/dL, 64% met their ATP III non-HDL goals. PMID- 12646345 TI - Some personal history and reflections from the life of a biochemist. PMID- 12646346 TI - John T. Edsall: an appreciation. PMID- 12646347 TI - Focal contributions to molecular biophysics and structural biology: a personal view. Part I. PMID- 12646348 TI - John Tileston Edsall: some selected biography and personal reminiscences. AB - This paper has been written in honor of John T. Edsall. Its purpose is to review briefly the status of science during his active years and the combination of personal qualities and upbringing which resulted in his enormous impact on the field of biochemistry. There was an unusual combination of deep knowledge and interest in many intellectual areas by no means restricted to science; the large number of individuals he met by intent or by chance, many to become fast friends for life, who themselves became distinguished in science; and last but not least his influence on science policy through very clear statements and his moral positions on questions of the day. PMID- 12646349 TI - John T. Edsall: his key role in the determination of the structure of proteins. AB - A letter, written in 1947 by John Edsall, outlined a declared intent to set up an X-ray crystallographic laboratory devoted to the study of crystalline heavy atom derivatives of proteins in an attempt to learn more about their structure. The fundamental idea, to the recipient (B.W.L.) totally new, revolutionary, and wholly contrary to all learned certainties, led to a correspondence, presented here in excerpt. Detailed plans were made for the laboratory to be built in the Department of Physical Chemistry at the Harvard Medical School. The proteins to be studied were reviewed and debated. The work of the laboratory is briefly described. Lack of success, the fatal consequence of a then unknown feature of the protein first chosen for study, is now only recently understood. The history of the Edsall idea and initiative is explored, from its beginnings to its acceptance and exploitation. John Edsall is here recognized as prime proponent and developer of the fundamental idea behind the most powerful and, for more than three decades, the only successful approach to the determination of protein structure. PMID- 12646350 TI - Reflections on a century of protein chemistry. AB - The development of protein structural chemistry during the twentieth century is briefly reviewed. Emphasis is placed on certain major problems that have defined the field, and how they have been resolved, often as a consequence of technological advances. The ways in which incorrect hypotheses have affected the development of the field are also discussed. PMID- 12646352 TI - John T. Edsall as tutor and teacher. PMID- 12646351 TI - 'Cohn and Edsall': physical chemistry conclusively supports a protein model. AB - The Harvard laboratory of Edwin J. Cohn and J.T. Edsall and the treatise on proteins that emerged from their work exerted a dominant effect from the 1920s to the 1950s. Its most important achievement was to solidify a molecular picture of proteins, especially the picture of a globular protein. This model representation did not originate with Cohn and Edsall, nor were they directly crusading for it. Their support for the model evolved simply from the sheer volume and diversity of their work. Everything they did or wrote about depended on a molecular picture. All of it was in the realm of physical chemistry and inexorably linked by rigorous theoretical equations to parameters of size, shape and ionic charge. PMID- 12646353 TI - My unusually dedicated mentor. PMID- 12646354 TI - Thinking about John T. Edsall. PMID- 12646355 TI - John Edsall's influence. PMID- 12646356 TI - Linus Pauling and sickle cell disease. AB - The 1949 paper by Linus Pauling et al. [Science 110 (1949) 543-548] describing the discovery of sickle cell anemia as the first molecular disease had a major impact on biology and medicine. Inspired by the scholarly works of John Edsall on the history of hemoglobin research, I present a brief retrospective analysis of Pauling's paper. This is followed by some personal recollections of Edsall and Pauling. PMID- 12646357 TI - John Edsall and ligand-linked subunit interactions in hemoglobin. PMID- 12646358 TI - Hemoglobin-oxygen equilibria: retrospective and phenomenological perspective. AB - The origin of the concept of a molecular complex between oxygen and hemoglobin can be traced to Stokes, a century and a half ago. Subsequently, physicochemical concepts of equilibria provided a path to quantitative formulations of these ligand-receptor interactions. Then, it took a quarter of a century before a proper format was prepared in terms of four stoichiometric equilibria and their associated binding constants. Since then, experimental measurements of these stoichiometric binding constants have consistently disclosed that successive values of K(1) to K(4) are accentuated above those expected if every subunit of hemoglobin maintained the same, intrinsic, unchanging affinity for oxygen. An alternative analysis of the observed cooperative interactions has been obtained by extracting the roots of the polynomial of the stoichiometric binding equation and deriving an alternative binding equation containing constants that for O(2) Hb are complex numbers. These constants have the dimensions and properties of equilibrium constants. They provide some novel phenomenological insights into ligand-receptor equilibria. PMID- 12646359 TI - Site mutations disrupt inter-helical H-bonds (alpha14W-alpha67T and beta15W beta72S) involved in kinetic steps in the hemoglobin R-->T transition without altering the free energies of oxygenation. AB - Three recombinant mutant hemoglobins (rHbs) of human normal adult hemoglobin (Hb A), rHb (alphaT67V), rHb (betaS72A), and rHb (alphaT67V, betaS72A), have been constructed to test the role of the tertiary intra-subunit H-bonds between alpha67T and alpha14W and between beta72S and beta15W in the cooperative oxygenation of Hb A. Oxygen-binding studies in 0.1 M sodium phosphate buffer at 29 degrees C show that rHb (alphaT67V), rHb (betaS72A), and rHb (alphaT67V, betaS72A) exhibit oxygen-binding properties similar to those of Hb A. The binding of oxygen to these rHbs is highly cooperative, with a Hill coefficient of approximately 2.8, compared to approximately 3.1 for Hb A. Proton nuclear magnetic resonance (NMR) studies show that rHb (alphaT67V), rHb (betaS72A), rHb (alphaT67V, betaS72A), and Hb A have similar quaternary structures in the alpha(1)beta(2) subunit interfaces. In particular, the inter-subunit H-bonds between alpha42Tyr and beta99Asp and between beta37Trp and alpha94Asp are maintained in the mutants in the deoxy form. There are slight perturbations in the distal heme pocket region of the alpha- and beta-chains in the mutants. A comparison of the exchangeable 1H resonances of Hb A with those of these three rHbs suggests that alpha67T and beta72S are H-bonded to alpha14W and beta15W, respectively, in the CO and deoxy forms of Hb A. The absence of significant free energy changes for the oxygenation process of these three rHbs compared to those of Hb A, even though the inter-helical H-bonds are abolished, indicates that these two sets of H-bonds are of comparable strength in the ligated and unligated forms of Hb A. Thus, the mutations at alphaT67V and betaS72A do not affect the overall energetics of the oxygenation process. The preserved cooperativity in the binding of oxygen to these three mutants also implies that there are multiple interactions involved in the oxygenation process of Hb A. PMID- 12646360 TI - Application of the continuous variation method to cooperative interactions: mechanism of Fe(II)-ferrozine chelation and conditions leading to anomalous binding ratios. AB - The method of continuous variation, often known as the Job plot, has long been used for determining the stoichiometry of two interacting components. The correct binding ratio, n, is generally obtained when the total concentration of the reactants, C(o), is much greater than the dissociation constants involved. For non-cooperative binding systems, the stoichiometry varies between one and n as C(o) increases; whereas for positive cooperative systems, values larger than n may be observed at low C(o). In this report, we present examples to illustrate how the changing apparent stoichiometries as a function of C(o) can provide clues for differentiating various binding mechanisms. To test these concepts, we examined the chelation of Fe(II) with ferrozine in the range of C(o)=7 to 210 microM with Fe(II) expressed in molar concentration or in terms of its binding equivalents (three in this case). The results were analyzed according to several models and found to be most consistent with the mechanism of one-step complex formation or infinite cooperativity with a K(d) of 8 microM. PMID- 12646361 TI - Dielectric behavior and atomic structure of serum albumin. AB - After 1946, serum albumin was available for studies. Its residue sequence and internal disulfide bonding was developed by 1976. We began to make dielectric dispersion studies and apply Perrin's equations for rotational relaxation times around the two axes of revolution in 1938. These data indicated that albumin should have an elongated shape. In 1992 atomic structure data indicated the molecule was heart-shaped. A similar 1998 study of albumin complexed with fatty acid showed that the molecule was substantially rearranged. We found that the dielectric constant of albumin solutions was sensitive to fatty acid content, making this property an attractive probe in stop-flow kinetic studies. Such studies show that the fatty acid reaction is a two-step process. The fatty acid first binds to exterior sites in a diffusion-limited second order reaction complete in 1 ms. Then a first order rearrangement reaction with approximately 400 ms half-life follows. Thus the highly specialized serum albumin sequence of amino acid residues determines not only the structure of the unligated molecule, but also the distinctive structures of the numerous multiligated molecules. PMID- 12646362 TI - Reflections on Edsall's carbonic anhydrase: paradoxes of an ultra fast enzyme. AB - John Edsall's investigations of human erythrocyte carbonic anhydrase, a zinc metalloenzyme that powerfully catalyzes the reversible hydration of carbon dioxide, highlighted a conundrum regarding the correct hydration product. The measured kinetic parameters could not be reconciled with the choice of carbonic acid, since its bimolecular recombination rate with enzyme would exceed the diffusion limit. The alternate choice of bicarbonate obviated the recombination rate problem but required that the active site deprotonation exceed the diffusion limited maximum rate by an even greater extent. This paradox was resolved in favor of bicarbonate when the unsuspected role of buffer species indirectly deprotonating the enzyme was finally proposed, spurring numerous investigations to verify the hypothesis. Edsall's laboratory also reported the accidental discovery of the first competitive inhibitor, imidazole. This opened new avenues to understanding the binding of the CO(2) substrate and stimulated many investigations on this inhibitor. Paramagnetic NMR and crystallographic studies demonstrated that the only other known competitive inhibitor, phenol, apparently shared this unusual binding site. Despite enormous progress since Edsall's retirement, particularly the use of site-directed mutagenesis approaches, the precise interactions of carbon dioxide and bicarbonate with specific active site moieties remain as elusive today as when Edsall first considered these questions. PMID- 12646363 TI - So near and yet so far from understanding molecular motors: recollections in honor of John T. Edsall. PMID- 12646364 TI - The impact of the paper of Edsall and Mehl (1940) on actin and actomyosin research. AB - The paper of Edsall and Mehl, 'The effect of denaturing agents on myosin, II. Viscosity and double refraction of flow', J. Biol. Chem. 133 (1940) 409-429, inspired our research on actin and actomyosin. It led to the specific purification of actin with magnesium ions and to the demonstration of the central role of the Mg(2+)-activated actomyosin ATPase in contraction of live muscle. PMID- 12646365 TI - The role of water in the thermodynamics of dilute aqueous solutions. AB - Water plays a role in the thermodynamics of dilute aqueous solutions that is unusual in two ways. First, knowledge of hydration equilibrium constants of species is not required in calculations of thermodynamic properties of biochemical reactants and reactions at specified pH. Second, since solvent provides an essentially infinite source of oxygen atoms in a reaction system where water is a reactant, oxygen atoms are not conserved in the reaction system in dilute aqueous solutions. This is related to the fact that H(2)O is omitted in equilibrium expressions for dilute aqueous solutions. Calculations of the standard transformed Gibbs energies of formation of total carbon dioxide and total ammonia at specified pH are discussed, and the average bindings of hydrogen ions by these reactants are calculated by differentiation. Since both of these reactants are involved in the urease reaction, the apparent equilibrium constants and changes in the numbers of hydrogen ions bound are calculated for this reaction as functions of pH. PMID- 12646366 TI - The basis of the hydrophobic effect. AB - The property of a molecule that most reliably determines the magnitude of the hydrophobic effect that it will experience is the number of hydrogen-carbon bonds it contains not the accessible surface area of its nonpolar portions. This conclusion follows from an examination of the standard free energies of transfer of alkanes, alkenes, alkadienes, and arenes from water to hexadecane. When the standard free energies of transfer for hydrocarbons in these different classes are plotted as a function of the number of hydrogen-carbon bonds they contain, all of the data fall upon the same line. These standard free energies of transfer are also directly proportional to the number of hydrogen-carbon bonds the hydrocarbons contain. When the same standard free energies of transfer are plotted as a function of the accessible surface areas of the hydrocarbons, the data do not fall upon the same line nor are the standard free energies of transfer directly proportional to the accessible surface areas. An examination of the standard free energies of transfer for the different classes of hydrocarbons from the gas phase to water and from the gas phase to hexadecane reinforces the conclusion that the number of hydrogen-carbon bonds in a molecule rather than its accessible surface area is the basis of the hydrophobic effect. Consequently, estimates of the contribution of different functional groups to the hydrophobic effect providing the free energy of folding of a molecule of protein or providing the free energy of dissociation for the association of two proteins or the association of a ligand with a protein should be made by counting the number of hydrogen-carbon bonds excluded from water rather than computing the accessible surface areas excluded from water. PMID- 12646367 TI - New considerations of the Barclay-Butler rule and the behavior of water dissolved in organic solvents. AB - The Barclay-Butler (B-B) rule, which states that a linear relationship exists between the standard deltaH(vap) and deltaS(vap) for simple, non-associated liquids and their solutions, has been used to distinguish associated ('abnormal') liquids from simple ('normal') liquids. The exact character of the B-B plots depends on the standard states chosen for the liquid/solution and vapor. We examine the effects of using number density for both vapor and liquid states for pure liquids, non-aqueous solutions, aqueous solutions and solutions in which water is the solute. The utility of B-B plots to detect solute-induced order is strengthened, and we also find remarkable changes in the modified B-B relationship: (1) the points for small, H-bonded liquids, including water, are pulled below the general B-B line; (2) many solutions containing small, simple solutes have negative entropies of vaporization; and (3) solutions of water in several organic solvents, relevant to studies of proteins and micelles, appear 'abnormal'. PMID- 12646368 TI - A microscopic view of peptide and protein solvation. AB - The structure and dynamics of the water hydrating peptides and proteins are examined here at atomic resolution via molecular dynamics simulations. Detailed solvation density and residence time data for all 20 L-amino acids in an end capped AXA tripeptide motif are presented. In addition, the solvation of the protein chymotrypsin inhibitor 2 is investigated as a point of comparison. Residues on the surface of proteins are not isolated; they interact both locally and non-locally in sequence space, and comparison of the solvation properties of each amino acid in both the peptide and protein allow us to distinguish inherent solvation properties from context-dependent perturbations due to neighboring residues. This work moves beyond traditional radial distribution functions and presents graphical representations of preferential solvation and orientation of water by side chains and the main chain. The combination of 0.3 micros of simulation data improves the statistical sampling over previous studies and reveals the significance of bridging water molecules that stabilize and mediate side chain-side chain, side chain-main chain and main chain-main chain interactions at the solvation interface. PMID- 12646369 TI - Group additivity schemes for the calculation of the partial molar heat capacities and volumes of unfolded proteins in aqueous solution. AB - A critical review is given of the present state of group additivity schemes for the calculation of partial molar volumes and heat capacities of unfolded proteins. The comparison between the experimental values and the predictions based on the different models shows clearly that only the peptide-based additivity scheme represents properly both the absolute values and the temperature dependence of these thermodynamic quantities. PMID- 12646371 TI - Modeling of electrostatic recognition processes in the mammalian mitochondrial steroid hydroxylase system. AB - Adrenodoxin reductase (AR) and adrenodoxin (Adx) are components of the mammalian mitochondrial steroid-hydroxylating system. Crystal structures of Adx, AR and a cross-linked Adx-AR complex have recently been determined. Based on these, we have carried out a modeling and docking study to characterize the recognition between AR, Adx and cytochrome c (Cytc). To rationalize the recognition process, electrostatic potentials were calculated by solving the Poisson-Boltzmann equations. In the Adx-AR complex modeled, a negatively charged surface of Adx recognizes a positive surface of AR, as in the crystal structure of the Adx-AR complex, proving the correct parameterization for the energy calculations. After forming salt bridges between the polar primary binding sites of Adx and AR, charge compensation causes a domain movement in AR, which closes the binding cleft by 2-4 A. Thereby, a secondary polar binding site is closed and the electron transfer pathways between the FAD of AR and the [2Fe-2S] cluster of Adx are adjusted. Next, the model structure of a complex between Adx and Cytc was derived. The lowest-energy complex between Adx and Cytc matches earlier chemical modification and cross-linking experiments, which proposed polar interactions of Lys13, Lys27, Lys72 and Lys79 of Cytc with acidic residues in Adx. Because of the short distance of 9.4 A between the redox centers, a complex, productive in electron transfer via a different outlet pathway from the inlet route in Adx, is expected. However, a ternary complex cannot be formed between the Adx-AR complex and Cytc because of steric hindrance. Therefore, a shuttle model for the role of Adx in the electron transfer process to Cytc is preferable to a relay model. In addition, no preferable docking site could be detected for a second Adx when probing the Adx-AR complex, which is required for a quaternary organized-cluster model of all redox partners of the hydroxylase system. PMID- 12646370 TI - Addition of side chains to a known backbone with defined side-chain centroids. AB - An automatic procedure is proposed for adding side chains to a protein backbone; it is based on optimization of a simplified energy function for peptide side chains, given its backbone and positions of side-chain centroids. The energy is expressed as a sum of the energies of interaction between side chains, and a harmonic penalty function accounting for the preservation of the positions of the C(alpha) atoms and the side-chain centroids. The energy of side-chain interactions is calculated with the soft-sphere ECEPP/3 potential. A Monte Carlo search is carried out to explore all possible side-chain orientations within a fixed backbone and side-chain centroid positions. The initial, usually extended, side-chain conformations are taken directly from the ECEPP/3 database. The procedure was tested on six experimental (X-ray or NMR) structures: immunoglobulin binding protein (PDB code 1IGD, an alpha+beta-protein); transcription factor PML (PDB code 1BOR, a 49-104 fragment of the ring finger domain, predominantly beta-protein); bovine pancreatic trypsin inhibitor (crystal form II) (PDB code 1BPI, an alpha+beta-protein); the monomer of human deoxyhemoglobin (PDB code 1BZ0, an alpha-helical structure); chain A of alcohol dehydrogenase from Drosophila lebanonensis (PDB code 1A4U); as well as on the 10 55 portion of the B domain of staphylococcal protein A (PDB code 1BDD). In all cases except 1BPI, the data for the algorithm (i.e. the backbone or C(alpha) coordinates and the positions of side-chain centroids) were taken from the experimental structures. For protein A, the C(alpha) coordinates and positions of side-chain centroids were also taken from the 1.9-A-resolution model predicted by the UNRES force field. In all comparisons with experimental structures, complete side-chain geometry was reconstructed with a root-mean-square (RMS) deviation of approximately 0.6-0.9 A from the heavy atoms when complete backbone and side chain-centroid coordinates were used in reconstruction, or approximately 1.0 A when the C(alpha) and centroid coordinates were used. PMID- 12646372 TI - Design and synthesis of peptides that bind alpha-bungarotoxin with high affinity and mimic the three-dimensional structure of the binding-site of acetylcholine receptor. AB - Alpha-bungarotoxin (alpha-BTX) is a highly toxic snake neurotoxin that binds to acetylcholine receptor (AChR) at the neuromuscular junction, and is a potent inhibitor of this receptor. In the following we review multi-phase research of the design, synthesis and structure analysis of peptides that bind alpha-BTX and inhibit its binding to AChR. Structure-based design concomitant with biological information of the alpha-BTX/AChR system yielded 13-mer peptides that bind to alpha-BTX with high affinity and are potent inhibitors of alpha-BTX binding to AChR (IC(50) of 2 nM). X-Ray and NMR spectroscopy reveal that the high-affinity peptides fold into an anti-parallel beta-hairpin structure when bound to alpha BTX. The structures of the bound peptides and the homologous loop of acetylcholine binding protein, a soluble analog of AChR, are remarkably similar. Their superposition indicates that the toxin wraps around the binding-site loop, and in addition, binds tightly at the interface of two of the receptor subunits and blocks access of acetylcholine to its binding site. The procedure described in this article may serve as a paradigm for obtaining high-affinity peptides in biochemical systems that contain a ligand and a receptor molecule. PMID- 12646373 TI - Some notes on crystallizing fibrinogen and fibrin fragments. AB - We have recently determined the structure of a native fibrinogen at 2.7-A resolution. Not the least of the hurdles during the many years of this project was growing X-ray-grade crystals from suitably purified proteins. Small, synthetic peptides based on the parts of fibrinogen exposed by the action of thrombin contributed greatly to these experiments. In addition, trimethylamine oxide (TMAO) was found to improve the diffraction of fibrinogen crystals. The history of my interest in fibrinogen and its crystallization can be traced back in part to some early interactions with John Edsall. PMID- 12646374 TI - Dissecting substrate recognition by thrombin using the inactive mutant S195A. AB - The catalytically inactive mutant S195A was used to study the interaction of thrombin with substrates under equilibrium conditions. By monitoring changes in intrinsic fluorescence, we measured dissociation constants for a variety of synthetic substrates, PAR peptides and the inhibitor PPACK. The S195A mutant retains the Na(+)-binding properties of the wild type, and substrate binding to the mutant is enhanced by the presence of Na(+). Temperature dependence studies allowed calculation of the thermodynamic parameters of substrate binding at the active site and showed a negligible deltaC(p). Titration of synthetic substrates carrying substitutions at the P1-P3 positions revealed energetics consistent with the specificity hierarchy identified in hydrolysis by the wild type. Titration with PAR peptides, which interact with both the active site and exosite I of thrombin, also showed consistency with the results obtained with the wild type at steady state. These findings demonstrate that inactive mutants of enzymes make it possible to dissect the equilibrium components linked to substrate binding and complement information on the kinetic properties of the wild type. PMID- 12646375 TI - Multiple methionine substitutions are tolerated in T4 lysozyme and have coupled effects on folding and stability. AB - In order to further explore the tolerance of proteins to amino acid substitutions within the interior, a series of core residues was replaced by methionine within the C-terminal domain of T4 lysozyme. By replacing leucine, isoleucine, valine and phenylalanine residues a total of 10 methionines could be introduced, which corresponds to a third of the residues that are buried in this domain. As more methionines are incorporated the protein gradually loses stability. This is attributed in part to a reduction in hydrophobic stabilization, in part to the increased entropic cost of localizing the long, flexible methionine sidechains, and in part to steric clashes. The changes in structure of the mutants relative to the wildtype protein are modest but tend to increase in an additive fashion as more methionines are included. In the most extreme case, namely the 10-methionine mutant, much of the C-terminal domain remains quite similar to wildtype (root mean-square backbone shifts of 0.56 A), while the F and G helices undergo rotations of approximately 20 degrees and center-of-mass shifts of approximately 1.4 A. For up to six methionine substitutions the changes in stability are additive. Beyond this point, however, the multiple mutants are somewhat more stable than suggested from the sum of their constituents, especially for those including the replacement Val111-->Met. This is interpreted in terms of the larger structural changes associated with this substitution. The substituted sidechains in the mutant structures have somewhat higher crystallographic thermal factors than their counterparts in WT*. Nevertheless, the interiors of the mutant proteins retain a well-defined structure with little suggestion of molten-globule characteristics. Lysozymes in which selenomethionine has been incorporated rather than methionine tend to have increased stability. At the same time they also fold faster. This provides further evidence that, at the rate-limiting step in folding, the structure of the C-terminal domain of T4 lysozyme is similar to that of the fully folded protein. PMID- 12646376 TI - Role of an N(cap) residue in determining the stability and operator-binding affinity of Arc repressor. AB - The Arc repressor of bacteriophage P22 is a member of the ribbon-helix-helix family of transcription factors. Ser32 is a solvent-exposed position that serves a structural role as the N(cap) residue of alpha-helix B of Arc, but also serves a functional role because its side chain is packed close to the sugar-phosphate DNA backbone in the repressor-operator complex. The tolerance of this N(cap) position to amino-acid substitutions was probed by determining the repressor activity in vivo, the thermal stability and the operator-binding activity in vitro of a set of 13 mutant proteins. The stability of position-32 Arc variants, except for Cys32, correlated well with the frequencies observed for the corresponding residues at N(cap) positions in alpha-helices of other proteins. Cysteine was quite stabilizing at the helix-B N(cap) position in Arc, but surprisingly was the least frequent N(cap) residue in the protein database. This latter finding may reflect a hyper-reactivity of N(cap) cysteines, which makes them prone to chemical modification. In general, only Arc variants with small, uncharged residues at position 32 were active in vivo or showed strong operator binding in vitro. Based upon the results presented here, revised sequence alignments of the MetJ and NikR subfamilies with Arc and other ribbon-helix-helix proteins are proposed. PMID- 12646377 TI - Thermodynamic characterization of the interaction of human cyclophilin 18 with cyclosporin A. AB - Isothermal titration calorimetry (ITC) was used to investigate thermodynamic parameters of the cyclosporin A (CsA)-cyclophilin 18 (hCyp18) association reaction. We have calculated the thermodynamic parameters (enthalpy, entropy, heat capacity, and free energy of binding) of the CsA/hCyp18 complexation. All but two methods described in the literature underestimate the affinity to hCyp18 of CsA. We found that the association constant (1.1.10(8) M(-1) at 10 degrees C) of CsA to hCyp18 is in close agreement with the reciprocal of the reported inhibitory constant of the peptidylprolyl cis/trans isomerase activity of hCyp18. Interpretation of the thermodynamic parameters in buffered solution of water, 30% glycerol and D(2)O leads to the conclusion that the highly specific binding of CsA to hCyp18 is mainly mediated through hydrogen bonding and to a lesser degree through hydrophobic interaction. Furthermore, the pH dependence of the association constant was determined and analyzed according to a single proton linkage model, resulting in a pK(a) value of 5.7 in free hCyp18 and below 4.5 in the CsA complexed form. Titration experiments using different single component buffers possessing different heats of ionization allowed us to estimate that statistically half a proton is transferred upon CsA binding from the binding interface of hCyp18 to the buffer at pH 5.5. No proton transfer was detected at pH 7.5. The thermodynamic results are discussed in relation to the published X ray and NMR structure of the free and CsA complexed hCyp18. PMID- 12646378 TI - Thermodynamics of protein folding: a microscopic view. AB - Statistical thermodynamics provides a powerful theoretical framework for analyzing, understanding and predicting the conformational properties of biomolecules. The central quantity is the potential of mean force or effective energy as a function of conformation, which consists of the intramolecular energy and the solvation free energy. The intramolecular energy can be reasonably described by molecular mechanics-type functions. While the solvation free energy is more difficult to model, useful results can be obtained with simple approximations. Such functions have been used to estimate the intramolecular energy contribution to protein stability and obtain insights into the origin of thermodynamic functions of protein folding, such as the heat capacity. With reasonable decompositions of the various energy terms, one can obtain meaningful values for the contribution of one type of interaction or one chemical group to stability. Future developments will allow the thermodynamic characterization of ever more complex biological processes. PMID- 12646379 TI - Non-linear rate-equilibrium free energy relationships and Hammond behavior in protein folding. AB - Non-linear rate-equilibrium relationships upon mutation or changes in solvent conditions are frequently observed in protein folding reactions and are usually interpreted in terms of Hammond behavior. Here we first give a general overview over the concept of transition state movements in chemical reactions and discuss its application to protein folding. We then show examples for genuine Hammond behavior and for apparent transition state movements caused by other effects like changes in the rate-limiting step of the folding reaction or ground state effects, i.e. structural changes in either the native state or the unfolded state. These examples show that apparent transition state movements can easily be mistaken for Hammond behavior. We describe experimental tests using self- and cross-interaction parameters to distinguish between structural changes in a single transition state following Hammond behavior and apparent transition state movements caused by other effects. PMID- 12646380 TI - Cytochrome c(551) as a model system for protein folding. AB - Considerable progress was made over the last few years in understanding the mechanism of folding of cytochrome c(551), a small acidic hemeprotein from Pseudomonas aeruginosa. Comparison of our results with those obtained by others on horse heart cytochrome c allows to draw some general conclusions on the structural features that are common determinants in the folding of members of the cytochrome c family. PMID- 12646381 TI - Native structural propensity in cellular retinoic acid-binding protein I 64-88: the role of locally encoded structure in the folding of a beta-barrel protein. AB - A central question in protein folding is the relative importance of locally encoded structure and cooperative interactions among residues distant in sequence. We have been exploring this question in a predominantly beta-sheet protein, since beta-structure formation clearly relies on both local and global sequence information. We present evidence that a 24-residue peptide corresponding to two linked hairpins of cellular retinoic acid-binding protein I (CRABP I) adopts significant native structure in aqueous solution. Prior work from our laboratory showed that the two turns contained in this fragment (turns III and IV) had the highest tendency of any of the eight turns in this anti-parallel beta barrel to fold into native turns. In addition, the primary sequence of these two turns is well conserved throughout the structural family to which CRABP I belongs, and residues in the turns and their associated hairpins participate in a network of conserved long-range interactions. We propose that the strong local sequence biases within the chain segment comprising turns III and IV favor longer range interactions that are crucial to the folding and native-state stability of CRABP I, and may play a similar role in related intracellular lipid-binding proteins (iLBPs). PMID- 12646383 TI - Annealing function of GroEL: structural and bioinformatic analysis. AB - The Escherichia coli chaperonin system, GroEL-GroES, facilitates folding of substrate proteins (SPs) that are otherwise destined to aggregate. The iterative annealing mechanism suggests that the allostery-driven GroEL transitions leading to changes in the microenvironment of the SP constitutes the annealing action of chaperonins. To describe the molecular basis for the changes in the nature of SP GroEL interactions we use the crystal structures of GroEL (T state), GroEL-ATP (R state) and the GroEL-GroES-(ADP)(7) (R" state) complex to determine the residue specific changes in the accessible surface area and the number of tertiary contacts as a result of the T-->R-->R" transitions. We find large changes in the accessible area in many residues in the apical domain, but relatively smaller changes are associated with residues in the equatorial domain. In the course of the T-->R transition the microenvironment of the SP changes which suggests that GroEL is an annealing machine even without GroES. This is reflected in the exposure of Glu386 which loses six contacts in the T-->R transition. We also evaluate the conservation of residues that participate in the various chaperonin functions. Multiple sequence alignments and chemical sequence entropy calculations reveal that, to a large extent, only the chemical identities and not the residues themselves important for the nominal functions (peptide binding, nucleotide binding, GroES and substrate protein release) are strongly conserved. Using chemical sequence entropy, which is computed by classifying aminoacids into four types (hydrophobic, polar, positively charged and negatively charged) we make several new predictions that are relevant for peptide binding and annealing function of GroEL. We identify a number of conserved peptide binding sites in the apical domain which coincide with those found in the 1.7 A crystal structure of 'mini-chaperone' complexed with the N-terminal tag. Correlated mutations in the HSP60 family, that might control allostery in GroEL, are also strongly conserved. Most importantly, we find that charged solvent-exposed residues in the T state (Lys 226, Glu 252 and Asp 253) are strongly conserved. This leads to the prediction that mutating these residues, that control the annealing function of the SP, can decrease the efficacy of the chaperonin function. PMID- 12646382 TI - Isolation and characterization of the prokaryotic proteasome homolog HslVU (ClpQY) from Thermotoga maritima and the crystal structure of HslV. AB - Heat-shock locus VU (HslVU) is an ATP-dependent proteolytic system and a prokaryotic homolog of the proteasome. It consists of HslV, the protease, and HslU, the ATPase and chaperone. We have cloned, sequenced and expressed both protein components from the hyperthermophile Thermotoga maritima. T. maritima HslU hydrolyzes a variety of nucleotides in a temperature-dependent manner, with the optimum lying between 75 and 80 degrees C. It is also nucleotide-unspecific for activation of HslV against amidolytic and caseinolytic activity. The Escherichia coli and T. maritima HslU proteins mutually stimulate HslV proteins from both sources, suggesting a conserved activation mechanism. The crystal structure of T. maritima HslV was determined and refined to 2.1-A resolution. The structure of the dodecameric enzyme is well conserved compared to those from E. coli and Haemophilus influenzae. A comparison of known HslV structures confirms the presence of a cation-binding site, although its exact role in the proteolytic mechanism of HslV remains unclear. Amongst factors responsible for the thermostability of T. maritima HslV, extensive ionic interactions/salt-bridge networks, which occur specifically in the T. maritima enzyme in comparison to its mesophilic counterparts, seem to play an important role. PMID- 12646385 TI - Lipid binding ability of human apolipoprotein E N-terminal domain isoforms: correlation with protein stability? AB - Human apolipoprotein (apo) E exists as one of three major isoforms, E2, E3 or E4. Individuals carrying the epsilon 4 allele have an increased risk of heart disease and premature onset of Alzheimer's disease. To investigate the molecular basis for this phenomenon, the N-terminal domain of apoE3, apoE2 and apoE4 were expressed in bacteria, isolated and employed in lipid binding and stability studies. Far UV circular dichroism spectroscopy in buffer at pH 7 revealed a similar amount of alpha-helix secondary structure for the three isoforms. By contrast, differences were noted in apoE-NT isoform-specific transformation of bilayer vesicles of dimyristoylphosphatidylglycerol (DMPG) into discoidal complexes. ApoE4-NT induced transformation was most rapid, followed by apoE3-NT and apoE2-NT. To determine if differences in the rate of apoE-NT induced DMPG vesicle transformation is due to isoform-specific differences in helix bundle stability, guanidine HCl denaturation studies were conducted. The results revealed that apoE2-NT was the most stable, followed by apoE3-NT and apoE4-NT, establishing an inverse correlation between helix bundle stability and DMPG vesicle transformation rate at pH 7. When the zwitterionic dimyristoylphosphatidylcholine (DMPC) was employed as the model lipid surface, interaction of apoE-NT isoforms with the lipid substrate was slow. However, upon lowering the pH from 7 to 3, a dramatic increase in the rate of DMPC vesicle transformation rate was observed for each isoform. To evaluate if the increased DMPC vesicle transformation rates observed at low pH is due to pH-dependent alterations in helix bundle stability, guanidine HCl denaturation studies were performed. ApoE2-NT and apoE3-NT displayed increased resistance to denaturation as a function of decreasing pH, while apoE4-NT showed no change in stability. Studies with the fluorescent probe, 8-anilino-1-naphthalene sulfonic acid, indicated an increase in apoE hydrophobic surface exposure upon decreasing the pH to 3.0. Taken together, the data indicate that changes in the stability of secondary structure elements in apoE-NT isoforms are not responsible for pH induced increases in lipid binding activity. It is likely that pH-induced disruption of inter-helical tertiary contacts may promote helix bundle conformational changes that present the hydrophobic interior of the protein to potential lipid surface binding sites. PMID- 12646384 TI - Physical-chemical features of non-detergent sulfobetaines active as protein folding helpers. AB - Some non-detergent sulfobetaines had been shown to prevent aggregation and improve the yield of active proteins when added to the buffer during in vitro protein renaturation. With the aim of designing more efficient folding helpers, a series of non-detergent sulfobetaines have been synthesized and their efficiency in improving the renaturation of a variety of proteins (E. coli tryptophan synthase and beta-D-galactosidase, hen lysozyme, bovine serum albumin, a monoclonal antibody) have been investigated. Attempts to correlate the structure of each sulfobetaines with its effect on folding revealed some molecular features that appear important in helping renaturation. This enabled us to design and synthesize new non-detergent sulfobetaines that act as potent folding helpers. PMID- 12646386 TI - Rapid chemical reaction techniques developed for use in investigations of membrane-bound proteins (neurotransmitter receptors). AB - New techniques for investigating chemical reactions on cell surfaces in the microsecond-to-millisecond time region are described. Reactions mediated by membrane-bound neurotransmitter receptors that control signal transmission between approximately 10(12) cells of the nervous system are taken as an example. Cells with receptors on their plasma membrane are equilibrated with photolabile, biologically inactive precursors of the neurotransmitters. Photolysis of these compounds releases free neurotransmitter that interacts with the receptors, leading to the transient opening of transmembrane receptor-formed channels that are permeant to small inorganic ions. The current thus induced can be measured. The technique can be used to measure the elementary steps of the receptor mediated reactions. To illustrate the approach it was shown that an understanding of the mechanism of inhibition of the nicotinic acetylcholine receptor by the drug cocaine was obtained and led to the first proof that compounds exist that alleviate the inhibition. PMID- 12646387 TI - Proteins, channels and crowded ions. AB - Ion channels are proteins with a hole down their middle that control a vast range of biological function in health and disease. Selectivity is an important biological function determined by the open channel, which does not change conformation on the biological time scale. The challenge is to predict the function-the current of ions of different types and concentrations through a variety of channels-from structure, given fundamental physical laws. Walls of ion channels, like active sites of enzymes, often contain several fixed charges. Those fixed charges demand counter ions nearby, and the density of those counter ions is very high, greater than 5 molar, because of the tiny volumes of the channel's pore. Physical chemists can now calculate the free energy per mole of salt solutions (e.g. the activity coefficient) from infinite dilution to saturation, even in ionic melts. Such calculations of a model of the L-type calcium channel show that the large energies needed to crowd charges into the channel can account for the substantial selectivity and complex properties found experimentally. The properties of such crowded charge are likely to be an important determinant of the properties of proteins in general because channels are nearly enzymes. PMID- 12646388 TI - Environmental influences on signal transduction through membranes: a retrospective mini-review. AB - This mini-review is addressed to the question how the membranous environment may affect traffic of receptors and signalling from membrane-anchored receptors on the outside of cells to transducers and targets on the inside. Signal transduction by membrane-anchored receptors to the interior of the cell and eventually to the genome is a central issue in cellular regulation. In this context the role of membrane fluidity and of the cytoskeleton in restricting the mobility of proteins are discussed and the evidence for a structural order in membranes which could limit the mobility of proteins is scrutinised. PMID- 12646389 TI - The significance of the free energy of hydrolysis of GTP for signal-transducing and regulatory GTPases. AB - A large number of GTP/GDP binding proteins, which in general have intrinsic and/or stimulatable GTPase activity, have been identified in recent years and are involved in a wide range of cellular regulatory and signal transducing processes. A common property of these proteins is that they exist in what is generally described as an active form when GTP is bound and an inactive (resting) form when GDP is present. Thus, the intrinsic or stimulated GTPase activity of these 'enzymes' serves to turn off a signal or to terminate a regulated process. It has been suggested that these proteins, together with ATPases whose prime function is to convert the free energy of ATP hydrolysis into another form of energy or into energy-requiring chemical reactions should be grouped together under the heading of 'energyases'. In this article, this suggestion is examined from the point of view of identifying the role of the free-energy of hydrolysis of GTP in the signal-transducing or regulatory process of the GTPases. It is concluded that there is a qualitative difference between ATPases and classical GTPases, in the sense that a quantitative relationship between the free-energy of GTP hydrolysis and the appearance of this energy in a different form cannot be directly defined. The significance of the high free energy of hydrolysis is that it allows efficient transition from the active to the inactive state of GTPases in spite of the tendency of the strong interaction of the GTP-bound form with a partner molecule ('effector'), an essential feature of their mode of action, to stabilize the GTP-bound form. PMID- 12646390 TI - The structural biology of growth factor receptor activation. AB - Stimulation of cells by growth factors triggers cascades of signalling that result in cellular responses such as growth, differentiation, migration and survival. Many growth factors signal through receptor tyrosine kinases, leading to dimerization, trans-phosphorylation and activation of tyrosine kinases that phosphorylate components further downstream of the signal transduction cascade. Using insulin-like growth factor, nerve growth factor, hepatocyte growth factor and fibroblast growth factor as examples, we show that the globular architecture of the growth factors is essential for receptor binding. We describe how nerve growth factor (NGF) is a symmetrical dimer that binds four storage proteins (two alpha-NGF and two gamma-NGF) to give a symmetrical hetero-hexameric 7SNGF organised around the beta-NGF dimer. It binds the extracellular domains of two receptor molecules in a similar way, so dimerising the receptor. Hepatocyte growth factor/scatter factor (HGF/SF) probably binds its receptor as a dimer stabilised by interactions with heparan sulfate, and fibroblast growth factor (FGF) binds its receptor as a dimer cross-linked by heparan sulfate. Surprisingly, insulin and insulin-like growth factor (IGF) bind in the monomeric form to receptors that are already covalent dimers. We propose that, in general, weak binary interactions between growth factor and individual domains of receptors are enhanced by cooperative interactions with further receptor domains, and sometimes other components like heparan, to give rise to specific multi protein/domain complexes. PMID- 12646391 TI - Transmembrane domains in the functions of Fc receptors. AB - In the present study, we use a novel method, PHDhtm, to predict the exact locations and extents of the transmembrane (TM) domains of multisubunit immunoglobulin Fc-receptors. Whereas most previous studies have used single residue hydrophobicity plots for characterizing of these domains, PHDhtm utilizes a system of neural networks and the evolutionary information contained in multiple alignments of related sequences to predict the above. Present PHDhtm application predicts TM domains of immunoglobulin Fc-receptors that in many cases differ significantly from those derived by using earlier methods. Comparisons of helical wheel projections of the presently derived TM domains from PHDhtm with those produced earlier reveal different hydrophobic moments as well as hydrophobic and hydrophilic surfaces. These differences probably alter the character of subunit association within the receptor complexes. This new algorithm can also be used for other membrane protein complexes and may advance both understanding the principles underlying such complexes formation and design of peptides that can interfere with such TM domain association so as to modulate specific cellular responses. PMID- 12646392 TI - Prospects of electron cryotomography to visualize macromolecular complexes inside cellular compartments: implications of crowding. AB - Electron cryotomography has unique potential for three-dimensional visualization of macromolecular complexes at work in their natural environment. This approach is based on reconstructing three-dimensional volumes from tilt series of electron micrographs of cells preserved in their native states by vitrification. Resolutions of 5-8 nm have already been achieved and the prospects for further improvement are good. Since many intracellular activities are conducted by complexes in the megadalton range with dimensions of 20-50 nm, current resolutions should suffice to identify many of them in tomograms. However, residual noise and the dense packing of cellular constituents hamper interpretation. Recently, tomographic data have been collected on vitrified eukaryotic cells (Medalia et al., Science (2002) in press). Their cytoplasm was found to be markedly less crowded than in the prokaryotes previously studied, in accord with differences in crowding between prokaryotic and eukaryotic cells documented by other (indirect) biophysical methods. The implications of this observation are twofold. First, complexes should be more easily identifiable in tomograms of eukaryotic cytoplasm. This applies both to recognizing known complexes and characterizing novel complexes. An example of the latter-a 5-fold symmetric particle is-given. Second, electron cryotomography offers an incisive probe to examine crowding in different cellular compartments. PMID- 12646393 TI - Reciprocal interactions between cells and extracellular matrix during remodeling of tissue constructs. AB - Cells remodel extracellular matrix during tissue development and wound healing. Similar processes occur when cells compress and stiffen collagen gels. An important task for cell biologists, biophysicists, and tissue engineers is to guide these remodeling processes to produce tissue constructs that mimic the structure and mechanical properties of natural tissues. This requires an understanding of the mechanisms by which this remodeling occurs. Quantitative measurements of the contractile force developed by cells and the extent of compression and stiffening of the matrix describe the results of the remodeling processes. Not only do forces exerted by cells influence the structure of the matrix but also external forces exerted on the matrix can modulate the structure and orientation of the cells. The mechanisms of these processes remain largely unknown, but recent studies of the regulation of myosin-dependent contractile force and of cell protrusion driven by actin polymerization provide clues about the regulation of cellular functions during remodeling. PMID- 12646394 TI - Quantitative approaches to problems of eukaryotic gene expression. AB - During the past several years there has been intense interest in the mechanisms by which gene expression is regulated within the eukaryotic nucleus. We have made use of an avian beta-globin locus to study various aspects of this problem, some of which are amenable to quantitative analysis. In the course of this work we have identified the transcription factor GATA-1, which is an essential regulatory factor for virtually all erythroid-specific genes, and studied the structure of its complex with its specific DNA binding site. The way in which GATA-1 forms tight interactions with DNA led to an understanding of how other zinc finger proteins of this class bind to DNA. We have extended such studies to examine interactions with DNA packaged as chromatin, and to studies of chromatin structure and function at higher levels of organization. PMID- 12646395 TI - On scientific freedom and responsibility. PMID- 12646396 TI - A novel RGD antagonist that targets both alphavbeta3 and alpha5beta1 induces apoptosis of angiogenic endothelial cells on type I collagen. AB - Integrin-mediated cell adhesion is necessary for endothelial cell proliferation and apoptosis, which is a major determinant in tumor-induced angiogenesis. In this study, we compared two novel, structurally similar, Arg-Gly-Asp (RGD) peptidomimetic compounds having different integrin selectivities, for their inhibition of endothelial cell proliferation and induction of apoptosis on functionally relevant extracellular matrices (ECM) for angiogenesis. BCH-14661 was specific for integrin alphavbeta3, whereas BCH-15046 nonselectively antagonized integrins alphavbeta3, alphavbeta5, and alpha5beta1. Both compounds were potent inducers of endothelial cell apoptosis when plated on RGD-dependent ECM (vitronectin, VN), which was dependent on the ability to induce cell detachment. However, with endothelial cells plated on RGD-independent ECM (type I collagen, COL), only BCH-15046 was able to significantly prevent growth and induce apoptosis. This effect was not dependent on the induction of detachment. Experiments using the matrix metalloproteinase (MMP) inhibitor GM 6001 revealed that cleavage of COL was not required for the ability of BCH-15046 to induce apoptosis. However, the inhibition of growth factor-stimulated endothelial cell proliferation, required MMPs, and correlated with BCH-15046s' potent inhibition of endothelial cell attachment to denatured collagen. Antibody inhibition experiments showed that adhesion to denatured collagen required integrins alphavbeta3 and beta1, but not alphavbeta5. In addition, BCH-15046 exerted a significant inhibition of VEGF-stimulated angiogenesis in the chick chorioallontoic membrane in vivo. These results suggest that integrin antagonism of both alphavbeta3 and alpha5beta1 are important for MMP-independent induction of apoptosis on COL and MMP-dependent inhibition of endothelial cell-denatured collagen interactions required for proliferation. PMID- 12646397 TI - Evidence for the role of alpha1D- and alpha1A-adrenoceptors in contraction of the rat mesenteric artery. AB - We investigated the alpha(1)-adrenoceptor subtype(s) involved in contraction of the isolated rat mesenteric artery by the use of the agonists noradrenaline (NA), phenylephrine (PHE), oxymetazoline (OXY), and methoxamine (MET), the competitive antagonists 8-(2-(4-(2-methoxyphenyl)-1-piperazinyl)ethyl)-8-azaspiro(4.5)decane 7,9-dione dihydrochloride (BMY 7378) and 5-methylurapidil, and the alkylating agent chloroethylclonidine (CEC). Agonists showed the potency order NA> or =PHE>OXY>MET; pA(2) values for 5-methylurapidil and BMY 7378 were 7.74+/-0.11 and 8.72+/-0.28, respectively, while Schild slopes were not different than unity; alpha(1)-adrenoceptor alkylation with CEC showed a drastic decrease in maximal agonists-induced contraction and a shift to the right of about 46-, 122-, 2-, and 15-fold higher than controls for NA, PHE, OXY, and MET, respectively. Data suggest that alpha(1D)-adrenoceptors predominate for contraction in mesenteric artery of the Wistar rat, with a second population of alpha(1A)-adrenoceptors responding at high agonist concentrations. PMID- 12646398 TI - In vivo and in vitro pharmacological studies of a new hypotensive compound (QF0301B) in rat: comparison with prazosin, a known alpha1-adrenoceptor antagonist. AB - In this work, we studied the in vivo and in vitro pharmacological effects of the novel compound QF0301B (2-[2-(N-4-o-methoxyphenyl-N-1-piperazinyl)ethyl]-1 tetralone) and compared with those of prazosin. In anaesthetized normotensive rats, both QF0301B and prazosin (0.1-0.2 mg/kg iv) caused a pronounced and prolonged fall in mean arterial blood pressure accompanied by bradycardia. Neither QF0301B nor prazosin (0.2 mg/kg iv) significantly modified the cardiovascular effects of either 5-hydroxytryptamine (serotonin, 5-HT, 75 microg/kg iv) or the selective alpha(2)-adrenoceptor agonist B-HT 920 (0.2 mg/kg iv), but both markedly inhibited the hypertensive effect of noradrenaline (5 microg/kg iv), a nonselective alpha-adrenergic receptor agonist. In isolated rubbed rat aorta rings, QF0301B and prazosin showed marked alpha(1)-adrenoceptor blocking activity, with pA(2) values of 9.00+/-0.12 and 9.75+/-0.14, respectively. In addition, QF0301B reversed and competitively antagonized the inhibitory action produced by clonidine in electrically stimulated rat vas deferens and inhibited the force and rate of contraction in rat isolated atria (pA(2)=5.91+/-0.43), competitively antagonized the contractile effect of 5-HT in rat aorta (pA(2)=6.75+/-0.06) and in rat stomach fundus (pA(2)=7.13+/-0.48) and the contractions induced by histamine in isolated guinea pig longitudinal ileal muscle (pA(2)=7.40+/-0.40). QF0301B showed noncompetitive low action in 5-HT(3), muscarinic and nicotinic receptors, or as Ca(2+) antagonist. These results indicate that a alpha(1)-adrenoceptor blocking lead has been obtained with a new chemical structure and interesting pharmacological properties, which only alpha(1)-adrenoceptor blocking activity seems to be responsible for its cardiovascular effects. PMID- 12646400 TI - Oxidative metabolism of peripheral blood neutrophils in experimental acute hypercapnia in the mechanically ventilated rabbit. AB - BACKGROUND AND AIM: The aim of this study was to investigate the effect of acute hypercapnia due to the mechanical hypoventilation on the oxidative metabolism of peripheral blood neutrophils in the rabbit. METHODS: The study was performed on 24 Chinchilla rabbits, randomized into normo- and hypercapnia groups (P(a)CO(2) between 9 and 11 kPa over 180 min). At the baseline point and after 180 min of mechanical ventilation, a neutrophil count, luminol-dependent chemiluminescence of the neutrophils stimulated with opsonized zymosan (OZ) or PMA (phorbol myristate acetate), and the hydrogen peroxide production of the cells upon the PMA stimulation were measured. Serum cortisol levels were additionally determined. RESULTS: In both studied groups, a significant neutrophilia after 180 min of mechanical ventilation was observed (P<.05). Neither chemiluminescence nor hydrogen peroxide production of peripheral neutrophils was changed (P>.05). CONCLUSIONS: Hypercapnia lasting for 180 min did not affect the oxidative metabolism of circulating neutrophils but a rise in a neutrophil count was observed in the mechanically ventilated rabbit. PMID- 12646399 TI - Role of hemin in the modulation of H2O2-mediated endothelial cell injury. AB - Heme oxygenase (HO) has been primarily regarded as the rate-limiting enzyme in the degradation of heme. However it has recently been proposed that the inducible isoform, HO-1 (EC 1.14.99.3), functions as a stress-responsive antioxidant enzyme, with the capacity to protect against oxidant-mediated vascular injury. This study used an in vitro model of endothelial permeability to determine the effects of the HO-1-inducing agent hemin on noncytotoxic endothelial injury mediated by acute oxidant stress. Effects of hemin on oxidant-mediated cytotoxicity in a number of endothelial cell types were also investigated. A 20 min exposure of human umbilical vein endothelial cell (HUVEC) monolayers to H(2)O(2) resulted in a significant concentration-dependent increase in permeability, which was reversible 48 h later. Pretreatment of monolayers with hemin for 2 h followed by 18 h in complete medium resulted in HO-1 induction and the attenuation of H(2)O(2)-mediated increases in endothelial permeability, and significantly improved the restoration of endothelial barrier function 48 h later. In HUVEC and in the human microvascular endothelial cell line HMEC-1, hemin treatment as above resulted in protection against cytotoxicity, but not in bovine aortic endothelial cells (BAECs), where such toxicity was potentiated. This potentiation was inhibited by incubation with the HO inhibitor tin protoporphyrin IX, supporting a role for HO-1 in the potentiation of the cytotoxic response. When the exposure time of BAEC to hemin was extended to 24 h, H(2)O(2)-mediated cytotoxicity was attenuated. We conclude that hemin treatment is cytoprotective against noncytotoxic endothelial injury in vitro, under conditions that may not offer global protection against cytotoxic injury to vascular endothelium. This would indicate that HO-1 induction associated with cytotoxic injury in vivo is not always beneficial and therefore that the use of hemin as a therapeutic agent to offset oxidant injury in vascular endothelium should be undertaken with caution. PMID- 12646401 TI - Characterization of alpha2-adrenoceptors in smooth muscles of the spontaneously hypertensive rat aorta. AB - Previous works have shown that the alpha(2)-adrenoceptor agonist UK 14,304 induced the relaxation and hyperpolarization of the rat aorta, mediated by alpha(2)-adrenoceptors present in the smooth muscles, through small-conductance, ATP-sensitive K(+) channels. We now report that in spontaneously hypertensive rat (SHR) aortic rings, UK 14,304 induced concentration-dependent hyperpolarizing responses, which were inhibited by yohimbine, an alpha(2)-adrenoceptor inhibitor, and by glibenclamide, a specific inhibitor of small-conductance, ATP-sensitive K(+) channels. The responses were also partially inhibited by iberiotoxin and by apamin. Treatment with N(omega)-nitro-L-arginine (L-NNA) did not affect the response to UK 14,304. These results indicate that alpha(2)-adrenoceptors are present in SHR aortic smooth muscle cell membranes, but differ from those of normotensive animals regarding the K(+) channels involved in their responses. Moreover, the resting membrane potential (RMP) was significantly more negative in SHR than in normotensive rats. This relative hyperpolarized state is probably due to Ca(2+)-dependent K(+) channels being constitutively open in SHR, since the addition of iberiotoxin caused a significant depolarization of the aortic smooth muscle membranes in this strain. PMID- 12646402 TI - Ventrally emigrating neural tube cells contribute to the normal development of heart and great vessels. AB - We investigated the contributions of a recently described population of neural tube cells, which participates in the development of a variety of tissues, to the development of the heart and great vessels. These cells, termed as the ventrally emigrating neural tube (VENT) cells, originate in the ventral part of the hindbrain neural tube, emigrate at the site of attachment of the cranial nerves, and populate their respective target tissues. VENT cells of the caudal hindbrain neural tube at the level of the vagus nerve, which were previously reported to migrate into the heart, were tagged with replication-deficient retroviruses containing the LacZ gene in chick embryos, after the emigration of neural crest from this region. In older embryos, VENT cells were detected in a variety of locations including the ventricles, atria, their septa, aorticopulmonary septum, and great vessels of the heart. Immunostaining with a specific marker revealed that VENT cells differentiated into smooth muscle cells of great vessels. Differentiation of VENT cells into cardiac muscle cells was reported previously. Extirpation of the VENT cells prior to their departure from the neural tube resulted in some common cardiovascular malformations: thin-walled ventricles and atria, ventricular and atrial septal defects, persistent truncus arteriosus, and stenosis of the great vessels. These results suggest that a novel population of neural tube cells also contributes to the normal development of the heart and great vessels. Thus, the heart and great vessels develop from three sources of cells: mesoderm, neural crest, and VENT cells. PMID- 12646403 TI - A fluctuation in adrenocepter- and muscarinic receptor-mediated blood pressure responses in acute hyperthyroid rats. AB - Hyperthyroidism was induced by daily subcutaneous injections of L-thyroxine (T(4), 0.5 mg/kg/day) for 3 days, 1 week, or 2 weeks to study whether there is a fluctuation in adrenoceptor- and muscarinic receptor-mediated blood pressure responses at a hyperthyroid stage. T(4) treatment for 3 days or 1 week significantly suppressed the pressor response induced by norepinephrine (NE). The depressor responses induced by isoprenaline or acetylcholine (ACh) were increased by T(4) treatment for only 3 days. The pressor response induced by N(G)-nitro-L arginine (L-NA) was increased by T(4) treatment for only 3 days. Results suggest that adrenoceptor- and muscarinic receptor-mediated blood pressure responses fluctuate in hyperthyroidism caused by T(4) in rats, that the basal nitric oxide (NO) production and/or release are increased in hyperthyroid rats at an early stage of the disease. PMID- 12646404 TI - Repeated constriction to respiration-induced hypocapnia is not mimicked by isocapnic alkaline solution in rabbit basilar artery in situ. AB - The purpose of this study was to test whether constriction of the cerebral vasculature in response to respiration-induced hypocapnia was mimicked by isocapnic alkaline solution. Since the regulation of the cerebral vasculature by hypocapnia necessitates vessels to constrict repeatedly in response to hypocapnic challenge, we tested whether repeated challenge with isocapnic alkaline solution was also associated with constriction. In contrast to our previous demonstration that repeated hypocapnic challenge elicited constrictions of similar magnitudes in rabbit basilar artery in situ, repeated challenge with isocapnic alkaline solution resulted in reduced constriction. Constriction to hypocapnia was also reduced following isocapnic alkaline solution. Since we previously demonstrated that constrictions to hypocapnia and isocapnic alkaline solution were endothelin 1 dependent, we tested whether the inhibition of hypocapnia- and isocapnic alkaline solution-induced constrictions following isocapnic alkaline solution was due to reduced endothelin-1 constriction. Endothelin-1 constriction was not reduced following isocapnic alkaline solution. Thus, constriction to isocapnic alkaline solution does not mimic constriction to hypocapnia. The results further suggest that the decreased constriction to isocapnic alkaline solution is due to blockade of endothelin-1 release, and that both hypocapnia and isocapnic alkaline solution share a common step in their endothelin-1 release pathways that can be inhibited by isocapnic alkaline solution. PMID- 12646405 TI - Effects of hydrazine derivatives on vascular smooth muscle contractility, blood pressure and cGMP production in rats: comparison with hydralazine. AB - Hydralazine is a hydrazine derivative used clinically as a vasodilator and antihypertensive agent. Despite numerous studies with the drug, its mechanism of action has remained unknown; guanylate cyclase activation and release of endothelial relaxing factors are thought to be involved in its vasodilator effect. Other hydrazine derivatives are known to stimulate guanylate cyclase and could therefore share the vasodilator activity of hydralazine, although such possibility has not been assessed systematically. In the present study, hydralazine, hydrazine, phenylhydrazine, and isoniazid were evaluated for vascular smooth muscle relaxation in rat aortic rings with and without endothelium, as well as after incubation with the guanylate cyclase inhibitor methylene blue. They were also tested for enhancement of cyclic guanosine monophosphate (cGMP) production by cultured rat aortic smooth muscle cells and for hypotension in the anesthetized rat. All hydrazines relaxed aortic rings, an action unaffected by endothelium removal and, in all cases except hydralazine, antagonized by methylene blue. Only phenylhydrazine increased cGMP production and only hydralazine markedly lowered blood pressure. It was concluded that hydralazine vascular relaxation is independent of endothelium and is not related to guanylate cyclase activation. The other hydrazines studied also elicit endothelium-independent relaxation, but the effect is related to guanylate cyclase. The marked hypotensive effect of hydralazine contrasts with its modest relaxant activity and is not shared by the other hydrazines. The fact that hydrazine and isoniazid produce methylene blue-sensitive relaxation, yet do not enhance cGMP production suggests the need for activating factors present in aortic rings but not in isolated cells. PMID- 12646406 TI - Functional evidence that K+ is the non-nitric oxide, non-prostanoid endothelium derived relaxing factor in rat femoral arteries. AB - The mechanisms of K(+)-induced relaxation and of acetylcholine (ACh)-stimulated, endothelium-dependent relaxation were assessed in rat femoral arteries mounted in a myograph. ACh-stimulated (1 nM-1 microM) relaxation of arteries precontracted with 1 microM noradrenaline was mostly resistant to the combination of indomethacin (INDO; 10 microM) and N(omega)-nitro-L-arginine (L-NNA, 100 microM). The remaining relaxation was abolished by 30 mM K(+) or ouabain (1 mM) and significantly reduced by 30 microM Ba(2+) or charybdotoxin (ChTx; 100 nM) plus apamin (100 nM). K(+)-induced relaxation effected by raising [K(+)](o) by 0.5-4 mM was endothelium-independent and inhibited by ouabain and Ba(2+). These results indicate that ACh-stimulated relaxations are effected mainly by a non-prostanoid, non-nitric oxide mechanism, presumably an endothelium-derived hyperpolarising factor (EDHF). Relaxations stimulated by EDHF and K(+) are both mediated by Na(+) K(+) ATPase and inward rectifier potassium channels (K(IR)). This study provides further functional evidence that EDHF is K(+) derived from endothelial cells that relaxes arterial smooth muscle subsequent to activation of Na(+)-K(+) ATPase and K(IR). PMID- 12646407 TI - The effects of nifedipine and thapsigargin on the responses of pressurized rat mesenteric artery to 5-hydroxytryptamine and norepinephrine. AB - The responses of isolated pressurized second order mesenteric resistance arteries of Wistar rats, superfused with physiological salt solution (PSS) were determined to 5-hydroxytryptamine (5-HT) and norepinephrine (NE). The contractility of the vessel was enhanced in response to 5-HT compared to NE (P<.001, ANOVA). The L type calcium ion channel blocker, nifedipine (10(-6) M) abolished the response to either 5-HT or NE. In vessels with intact endothelium, thapsigargin (TG, 10(-6) M), which inhibits uptake of calcium ions into intracellular stores, significantly reduced the contractile response to 5-HT (P<.02) but had little or no effect on the response to NE (P=.2). However, in vessels denuded of the endothelium, there was no significant difference in the response of the mesenteric artery, after TG, to either 5-HT or NE. The results indicate that, in the rat mesenteric resistance vessel, both 5-HT and NE use calcium ions from extracellular sources for contraction, while NE relies mainly on extracellular ion influx with little or no contribution from intracellular sources. The reduced response of the de-endothelized vessel to 5-HT after TG suggests that the utilization of intracellular stores by this agonist is endothelium-dependent. These observations may explain the enhanced responsiveness of the mesenteric artery to 5-HT when compared with NE. PMID- 12646408 TI - Pramanicin, an antifungal agent, raises cytosolic Ca2+ and causes cell death in vascular endothelial cells. AB - The effects of a newly discovered antifungal agent, pramanicin, on cytosolic Ca(2+) and cell viability of cultured bovine pulmonary artery endothelial cells and on endothelium-dependent relaxation of dog carotid arterial rings were investigated by digital dynamic fluorescence ratio imaging and morphological and contractility studies, respectively. Pramanicin 100 microM, previously shown to cause maximal endothelium-dependent and NO-mediated vascular relaxation, induced a small transient elevation of cytosolic Ca(2+) concentration in Ca(2+)-free medium; subsequent introduction of 1 mM Ca(2+) caused a steady, nonsaturating increase of Ca(2+), which could be brought down to the basal level by the addition of EGTA. At the single cell level, the elevation of cytosolic Ca(2+) initiates from the cell periphery and progresses toward the central region. When added to the plateau phase of phenylephrine-induced contraction, pramanicin induced a slow endothelium-dependent relaxation, which could be reversed with the NO synthase inhibitor, L-NOARG. When preincubated with vascular tissue, pramanicin resulted in an irreversible loss of endothelial function characterized by the lack of carbachol-induced relaxation. Pramanicin caused cell injury characterized by plasmalemmal bleb formation, leading to cell death characterized by Trypan blue staining of the nuclei in cultured vascular endothelial cells in a concentration- and time-dependent manner. Such pramanicin-induced cell death was not associated with Ca(2+)-mediated or NO-mediated mechanisms. The time course of Ca(2+) elevation corresponds with that of pramanicin-induced relaxation of precontracted arterial rings, whereas the time course of endothelial cell death corresponds to that of pramanicin-induced loss of endothelial function as assessed by carbachol-induced relaxation. The pramanicin analogue, PMC-A, a by product of the biosynthesis of pramanicin, in which the epoxy group is replaced by a CC bond, caused little endothelial-dependent relaxation, but it was able to cause endothelial cell dysfunction, albeit to a lesser extent compared to pramanicin, suggesting a role of the epoxy group in pramanicin for its vasorelaxant effect. PMID- 12646409 TI - Sympathetic stimulation of renin is independent of direct regulation by renal nitric oxide. AB - Nitric oxide (NO) regulates renin secretion through various pathways. The possibility that renal neuronal nitric synthase (nNOS) may mediate beta adrenergic control of renin was tested. In six Inactin-anesthetized rats, renin secretion rate (RSR) was measured in response to the beta-agonist isoproterenol with and without selective inhibition of nNOS using 7-nitroindazole (7-NI, 50 mg/kg body weight [BW]). 7-NI had no effect on blood pressure (BP) or renal hemodynamics, while isoproterenol increased RSR by 9 ng AngI/h/min (P<.05) similarly with or without 7-NI. Isoproterenol decreased BP by 20 mm Hg (P<.001), but this depressor response was completely blocked by 7-NI. When acute endogenous stimulation of renal sympathetic nerve activity (RSNA) was induced by bilateral carotid occlusion, BP in 12 rats (105+/-5 mm Hg) rose transiently to peak at 121+/-6 mm Hg (P<.005) within 5 min, returning to baseline within 10 min. RSR rose threefold (2.1+/-0.5 to 7.6+/-3.3 ng AngI/ml/min/g kidney weight [KW]; P<.05). Next, 7-NI had no effect on BP (108+/-5 mm Hg), but subsequent carotid occlusion increased and sustained BP by 27+/-5 mm Hg (P<.001), but RSR did not change (2.46+/-0.94 ng AngI/ml/min/g KW). However, if after 7-NI treatment followed by carotid occlusion, the renal perfusion pressure was not allowed to rise, but held constant at 111+/-3 mm Hg, RSR increased from 3.03+/-0.79 to 12.97+/-3.41 ng AngI/ml/min/g KW (P<.025). Thus, neither beta-adrenergic stimulation of RSR with isoproterenol nor direct stimulation of RSR by activation of RSNA with carotid occlusion was modified by selective nNOS inhibition. These data suggest an important nNOS component in the regulation of BP in response to carotid occlusion, but do not support a direct role of renal nNOS mediating sympathetic regulation of RSR. PMID- 12646410 TI - Tert-butyl hydroperoxide-mediated vascular responses in DOCA-salt hypertensive rats. AB - tert-Butyl hydroperoxide (t-BOOH), a membrane permeant oxidant, elicits enhanced vasoconstriction of perfused kidney and mesenteric arterial beds isolated from DOCA-salt-induced hypertensive rats. We hypothesize that enhanced vasoconstriction to t-BOOH during DOCA-salt hypertension involves free radical species and decreases in the expression of the endogenous antioxidant enzyme, superoxide dismutase (SOD). t-BOOH (0.01-50 micromol) dose-dependently constricted the perfused kidney and mesenteric vascular beds (MVB) of rats. Infusion of tempol (100 microM), a free radical scavenger, reduced the constrictor responses from 116.70+/-16.65% to 57.45+/-9.25% (kidneys) and from 72.91+/-3.70% to 48.10+/-0.10% (mesenteric beds). t-BOOH-induced vasoconstriction of both vascular beds were also significantly reduced in DOCA-salt rats treated chronically (15 mg/kg ip, 3 weeks) with tempol (DOCA/TEMPOL). Catalase (500 IU) did not attenuate t-BOOH-induced responses in vascular beds of DOCA/TEMPOL rats. Western blot analyses showed significant reduction in Cu/Zn-SOD expression in DOCA-salt versus sham rats of both vascular preparations; SOD expressions were protected from down-regulation in DOCA/TEMPOL vascular beds. This study suggests that free radical species is involved in both t-BOOH-induced constrictions and in the down-regulation of SOD protein expressions during DOCA-salt hypertension. PMID- 12646411 TI - Vasorelaxant effects of eugenol on rat thoracic aorta. AB - Eugenol is a natural pungent substance and the main component of clove oil, with vasorelaxant action. To elucidate some of the possible mechanisms involved in this action isometric tension was measured in aortic rings from male Wistar rats precontracted with phenylephrine (PHE, 10(-7) M) or KCl (75 mM). Responses to increasing concentrations of eugenol (10(-6)-10(-2) M) were obtained in the presence and absence of endothelium. In the presence of eugenol, dose-response curves to PHE (10(-9) to 10(-4) M) and KCl (5-125 mM) were displaced downwards. Concentration-dependent relaxation was observed in rings precontracted with PHE (10(-7) M) and KCl (75 mM). The tension increment produced by increasing external calcium concentration (0.25-3 mM) was also reduced by eugenol (300 microM) treatment. The inhibitory effects of eugenol (300 microM) were compared to those induced by nifedipine (0.01 microM), a selective Ca(2+) channel blocker, producing similar relaxant effects. Two other protocols were performed. After precontraction with PHE (10(-7) M), increasing concentrations of eugenol (10(-6) 10(-2) M) were used before and after N(w)-nitro-L-arginine (L-NAME, 10(-4) M) and methylene blue (10(-5) M) treatment. Eugenol-induced relaxation was reduced by endothelial damage (rubbing), L-NAME and methylene blue treatments. Results suggested that eugenol produces smooth muscle relaxation resulting from the blockade of both voltage-sensitive and receptor-operated channels that are modulated by endothelial-generated nitric oxide. PMID- 12646412 TI - Time-dependent hyperreactivity to phenylephrine in aorta from untreated diabetic rats: role of prostanoids and calcium mobilization. AB - Diabetes alters vascular smooth muscle contractility. Changes in reactivity to phenylephrine (Phe) in aortas from controls and untreated 1- and 4-week streptozotocin (STZ)-induced diabetic rats were investigated. In 1-week diabetic (DB1) aortas, the maximum response (E(max)) and sensitivity (pD(2)) to Phe were similar to controls (CT1), but in 4-week diabetic (DB4) aortas, the E(max) for Phe was increased compared to CT4 aortas (E(max), DB4: 125+/-8.4% vs. CT4: 89.8+/ 4.5%, P<.001). Endothelial denudation increased the response to Phe, and E(max) was increased in the DB4 aortas compared to CT4 (E(max), DB4: 156+/-4.2% vs. CT4: 125+/-3.8%, P<.001). Pretreatment of CT4 and DB4 aortas with indomethacin reduced E(max) and pD(2) for Phe. After indomethacin treatment, no differences in E(max) and pD(2) to Phe were observed in either group. SQ 29548 did not alter the Phe actions in CT4 aortas. However, in DB4 aortas, E(max) was reduced to control level. CT4 and DB4 aortas incubated in free-Ca(2+) solution plus Phe, contracted upon addition of CaCl(2), this response was increased in DB4 aortas. No changes were observed for acetylcholine (ACh) or sodium nitroprusside (SNP) responses. Nitric oxide (NO) release in response to Phe determined by acute L-NAME administration showed no differences in the percentage increase of the contraction in CT1 and DB1 aortas, but was enhanced in DB4 aortas. Results suggested that diabetes induces time-dependent changes in the vascular reactivity to Phe. This response is not related to a reduction of endothelium-derived NO but might be due to an increase in prostaglandin H(2) (PGH(2))/thromboxane A(2) (TxA(2)) and/or an enhanced extracellular Ca(2+) influx. PMID- 12646413 TI - Skeletal muscle sarcoplasmic reticulum contains a NADH-dependent oxidase that generates superoxide. AB - Skeletal muscle sarcoplasmic reticulum (SR) is shown to contain an NADH-dependent oxidase (NOX) that reduces molecular oxygen to generate superoxide. Its activity is coupled to an activation of the Ca2+ release mechanism, as evident by stimulation in the rate of high-affinity ryanodine binding. NOX activity, coupled to the production of superoxide, is not derived from the mitochondria but is SR in origin. The SR preparation also contains a significant NADH oxidase activity, which is not coupled to the production of superoxide and appears to be mitochondrial in origin. This mitochondrial component is preferentially associated with the terminal cisternae region of the SR. Its activity is inhibited by diphenylene iodonium (10 microM), antimycin A (200 nM), and rotenone (40 nM) but is not coupled to the generation of superoxide or the stimulation of the ryanodine receptor. The rate of superoxide production per milligram of protein is larger in SR than in mitochondria. This NOX may be a major source of oxidative stress in muscle. PMID- 12646414 TI - Functional properties of a brain-specific NH2-terminally spliced modulator of Kv4 channels. AB - Kv4/K channel-interacting protein (KChIP) potassium channels are a major class of rapidly inactivating K channels in brain and heart. Considering the importance of alternative splicing to the quantitative features of KChIP gating modulation, a previously uncharacterized splice form of KChIP1 was functionally characterized. The KChIP1b splice variant differs from the previously characterized KChIP1a splice form by the inclusion of a novel amino-terminal region that is encoded by an alternative exon that is conserved in mouse, rat, and human genes. The expression of KChIP1b mRNA was high in brain but undetectable in heart or liver by RT-PCR. In cerebellar tissue, KChIP1b and KChIP1a transcripts were expressed at nearly equal levels. Coexpression of KChIP1b or KChIP1a with Kv4.2 channels in oocytes slowed K current decay and destabilized open-inactivated channel gating. Like other KChIP subunits, KChIP1b increased Kv4.2 current amplitude and KChIP1b also shifted Kv4.2 conductance-voltage curves by -10 mV. The development of Kv4.2 channel inactivation accessed from closed gating states was faster with KChIP1b coexpression. Deletion of the novel amino-terminal region in KChIP1b selectively altered the subunit's modulation of Kv4.2 closed inactivation gating. The role of the KChIP1b NH2-terminal region was further confirmed by direct comparison of the properties of the NH2-terminal deletion mutant and the KChIP1a subunit, which is encoded by a transcript that lacks the novel exon. The features of KChIP1b modulation of Kv4 channels are likely to be conserved in mammals and demonstrate a role for the KChIP1 NH2-terminal region in the regulation of closed inactivation gating. PMID- 12646415 TI - Transforming growth factor-beta inhibition of proteasomal activity: a potential mechanism of growth arrest. AB - Although the proteasome plays a critical role in the controlled degradation of proteins involved in cell cycle control, the direct modulation of proteasomal function by growth regulatory signaling has not yet been demonstrated. We assessed the effect of transforming growth factor (TGF)-beta, a potent inhibitor of cell growth, on proteasomal function. TGF-beta selectively decreased hydrolysis of the proteasomal substrate Cbz-Leu-Leu-Leu-7-amido-4-methyl-coumarin (z-LLL-AMC) in a concentration-dependent manner but did not inhibit hydrolysis of other substrates Suc-Leu-Leu-Val-Tyr-AMC (suc-LLVY-AMC) or Cbz-Leu-Leu-Glu-AMC (z LLE-AMC). An increase in intracellular oxidative injury occurred during incubation with TGF-beta. Furthermore, in vitro hydrolysis of z-LLL-AMC, but not suc-LLVY-AMC, was decreased by hydrogen peroxide. TGF-beta did not increase cellular expression of heat shock protein (HSP)90, a potent inhibitor of z-LLL AMC hydrolysis in vitro. The physiological relevance of TGF-beta inhibition of proteasomal activity was studied by assessing the role of z-LLL-AMC hydrolysis on cyclin-dependent kinase inhibitor expression and cell growth. TGF-beta increased expression of p27KIP1 but did not alter expression of p21WAF1 or p16INK4A. The peptide aldehyde Cbz-Leu-Leu-leucinal (LLL-CHO or MG132) potently inhibited z-LLL AMC hydrolysis in cell extracts as well as increasing p27KIP1 and decreasing cell proliferation. Thus growth inhibition by TGF-beta decreases a specific proteasomal activity via an HSP90-independent mechanism that may involve oxidative inactivation or modulation of proteasomal subunit composition and results in altered cellular expression of key cell cycle regulatory proteins such as p27KIP1. PMID- 12646416 TI - Decrease in activity of smooth muscle L-type Ca2+ channels and its reversal by NF kappaB inhibitors in Crohn's colitis model. AB - We investigated the mechanisms of dysmotility of the colonic circular muscle of the Crohn's disease rat model. Contractions induced by KCl, carbachol, and Bay K 8644 were decreased in circular smooth muscles isolated from 2,4,6 trinitrobenzenesulfonic acid (TNBS)-induced colitis rat colon. However, the absolute force and Ca2+ sensitivity of contractile proteins were not affected as assessed in alpha-toxin permeabilized smooth muscle. The current density of the L type Ca2+ channel in circular smooth muscle cells was significantly decreased in the TNBS-treated colonic cells. However, expressions of the L-type Ca2+ channel mRNA and protein did not differ between control and TNBS-treated preparations. Pretreatment with the NF-kappaB inhibitors pyrrolidinedithiocarbamate and sulfasalazine partially recovered the decreased contractility and current density of the L-type Ca2+ channel by TNBS treatment. These results suggest that the decrease in the contraction of circular smooth muscle isolated from TNBS-induced colitis rat colon, which may be related to gut dysmotility in Crohn's disease, is attributable to the decreased activity of the L-type Ca2+ channel. The dysfunction of the L-type Ca2+ channel may be mediated by NF-kappaB-dependent pathways. PMID- 12646417 TI - Biotin uptake by human intestinal and liver epithelial cells: role of the SMVT system. AB - It has been well established that human intestinal and liver epithelial cells transport biotin via an Na+-dependent carrier-mediated mechanism. The sodium dependent multivitamin transport (SMVT), a biotin transporter, is expressed in both cell types. However, the relative contribution of SMVT toward total carrier mediated uptake of physiological (nanomolar) concentrations of biotin by these cells is not clear. Addressing this issue is important, especially in light of the recent identification of a second human high-affinity biotin uptake mechanism that operates at the nanomolar range. Hence, we employed a physiological approach of characterizing biotin uptake by human-derived intestinal Caco-2 and HepG2 cells at the nanomolar concentration range. We also employed a molecular biology approach of selectively silencing the endogenous SMVT of these cells with specific small interfering RNAs (siRNAs), then examining carrier-mediated biotin uptake. The results showed that in both Caco-2 and HepG2 cells, the initial rate of biotin uptake as a function of concentration over the range of 0.1 to 50 nM to be linear. Furthermore, we found that the addition of 100 nM unlabeled biotin, desthiobiotin, or pantothenic acid to the incubation medium had no effect on the uptake of 2.6 nM [3H]biotin. Pretreatment of Caco-2 and HepG2 cells with SMVT specific siRNAs substantially reduced SMVT mRNA and protein levels. In addition, carrier-mediated [3H]biotin (2.6 nM) uptake by Caco-2 and HepG2 cells was severely (P 0.01) inhibited by the siRNAs pretreatment. These results demonstrate that the recently described human high-affinity biotin uptake system is not functional in intestinal and liver epithelial cells. In addition, the results provide strong evidence that SMVT is the major (if not the only) biotin uptake system that operates in these cells. PMID- 12646418 TI - HNF-1alpha and endodermal transcription factors cooperatively activate Fabpl: MODY3 mutations abrogate cooperativity. AB - Hepatocyte nuclear factor (HNF)-1alpha plays a central role in intestinal and hepatic gene regulation and is required for hepatic expression of the liver fatty acid binding protein gene (Fabpl). An Fabpl transgene was directly activated through cognate sites by HNF-1alpha and HNF-1beta, as well as five other endodermal factors: CDX-1, C/EBPbeta, GATA-4, FoxA2, and HNF-4alpha. HNF-1alpha activated the Fabpl transgene by as much as 60-fold greater in the presence of the other five endodermal factors than in their absence, accounting for up to one half the total transgene activation by the group of six factors. This degree of synergistic interaction suggests that multifactor cooperativity is a critical determinant of endodermal gene activation by HNF-1alpha. Mutations in HNF-1alpha that result in maturity onset diabetes of the young (MODY3) provide evidence for the in vivo significance of these synergistic interactions. An R131Q HNF-1alpha MODY3 mutant exhibits complete loss of synergistic activation in concert with the other endodermal transcription factors despite wild-type transactivation ability in their absence. Furthermore, whereas wild-type HNF-1alpha exhibited pairwise cooperative synergy with each of the other five factors, the R131Q mutant could synergize only with GATA-4 and C/EBPbeta. Selective loss of synergy with other endodermal transcription factors accompanied by retention of native transactivation ability in an HNF-1alpha MODY mutant suggests in vivo significance for cooperative synergy. PMID- 12646419 TI - Role of 5-HT1B/D receptors in canine gastric accommodation: effect of sumatriptan and 5-HT1B/D receptor antagonists. AB - The 5-HT1B/D receptor agonist sumatriptan has been proposed to treat dyspeptic symptoms, because it facilitates gastric accommodation. It is unknown whether stimulation of 5-HT1B/D receptors is involved. Thus, in four conscious dogs, we compared the effects of sumatriptan alone or combined with N-[4-methoxy-3-(4 methyl-1-piperazinyl) phenyl]-2'-methyl-4'-(5-methyl-1,2,4-oxadiazol-3-yl)-[1,1 biphenyl]-4-carboxamide hydrocloride (GR-127935), N-[3-[3 (dimethylamino)-ethoxy] 4-methoxyphenyl]-2'-[methyl-4'-(5-methyl-1,2,4-oxadiazol-3-yl)]-[1,1-biphenyl]-4 carboxamide hydrocloride (SB-216641 hydrochloride), or 3-[4-(4-chloro phenyl)piperazin-1-yl]-1,1-diphenyl-2-propanol hydrochloride (BRL-15572 hydrochloride) (respectively, nonselective 5-HT1B/D, selective 5-HT1B, and selective 5-HT1D receptor antagonists) on gastric accommodation to isobaric distensions performed with a barostat. An exponential and a linear model were used to fit the pressure-volume relationship. An exponential equation fitted the data better than a linear equation. Sumatriptan (800 nmol/kg iv) induced an immediate gastric relaxation (Deltavolume: 112 +/- 44 ml, P < 0.05). After sumatriptan, the pressure-volume curve was shifted toward significantly higher volumes. This effect was fully reversed by GR-127935 or SB-216641 but not by BRL 15572. In conclusion, 5-HT1B receptors seem to play an important role in modulating gastric accommodation to a distending stimulus. An exponential model for pressure-volume curves fits well with the concept of gastric adaptive relaxation. PMID- 12646420 TI - Regulation and role of p21 and p27 cyclin-dependent kinase inhibitors during hepatocyte differentiation and growth. AB - Unlike a large number of cell types that undergo terminal differentiation associated with permanent withdrawal from the cell cycle, mature quiescent hepatocytes retain high proliferative potential. We report here a specific behavior of members of the Cip/Kip family of cyclin-dependent kinase (Cdk) inhibitors during development of the rat liver and proliferation of normal hepatocytes. Expression of p21, p27, and p57 transcripts and proteins was downregulated during the differentiation process to low or undetectable levels in adult liver. In contrast to p27, p21 protein increased in a mitogen-dependent manner in isolated hepatocytes and its expression pattern correlated with that of cyclin D1. In proliferating hepatocytes, p21 was predominantly associated with cyclin D1, these proteins were colocalized in the nucleus and p21-associated retinoblastoma protein (pRb) kinase activity increased in parallel with that of cyclin D1. Overexpression of p21 in mitogen-stimulated hepatocytes reduced DNA synthesis. In contrast, inhibition of p21 expression by antisense or small interfering RNAs oligonucleotides accelerated S phase entry. Finally, expression of p21 and cyclin D1, but not p27 proteins was regulated by MAPK kinase/extracellular signal-regulated kinase and phosphatidylinositol 3-kinase ferric-reducing ability power/mammalian target of rapamycin signal transduction pathways. In conclusion, these results demonstrate a specific and differential regulation of p21 and p27 during hepatocyte differentiation and proliferation that may contribute to the control of quiescent differentiated hepatic cell proliferating activity. PMID- 12646421 TI - Inducible nitric oxide synthase is involved in acid-induced gastric hyperemia in rats and mice. AB - The role of different isoforms of nitric oxide synthase (NOS) in the gastric mucosal hyperemia, induced by 155 mM luminal hydrochloric acid (pH approximately 0.8) without a barrier breaker, was investigated. Rats were anesthetized with Inactin (120 mg/kg ip), and mice were anesthetized with Forene (2.2% in 40% oxygen gas at 150 ml/min); the gastric mucosa was exteriorized. Gastric mucosal blood flow was measured with laser-Doppler flowmetry (LDF) in rats treated with Nomega-nitro-l-arginine (l-NNA; unspecific NOS inhibitor), l-N6-(1 iminoethyl)lysine [l-NIL; inducible (i) NOS inhibitor], or S-methyl-l thiocitrulline [SMTC; neuronal (n) NOS inhibitor], 10 mg/kg, followed by 3 mg. kg 1. h-1 iv, in iNOS-deficient (-/-) and nNOS(-/-) mice. mRNA was isolated from the gastric mucosa in iNOS(-/-) and wild-type (wt) mice, and real-time RT-PCR was performed. The effect of 155 mM acid on gastric mucosal permeability was determined by measuring the clearance of 51Cr-EDTA from blood to lumen. LDF increased by 48 +/- 13% during 155 mM HCl luminally, an increase that was abolished by l-NNA, SMTC, or l-NIL. In iNOS wt mice, LDF increased by 33 +/- 8% during luminal acid. The blood flow increase was attenuated substantially in iNOS(-/-) mice. RT-PCR revealed iNOS mRNA expression in the gastric mucosa in the iNOS wt groups. The blood flow increase in response to acid was not abolished in nNOS(-/-) mice (nNOS-sufficient mice, 39 +/- 18%; heterozygous mice, 25 +/- 19%; /- mice, 19 +/- 7%). Mucosal permeability was transiently increased during 155 mM HCl. The results suggest that iNOS is constitutively expressed in the gastric mucosa and is involved in acid-induced hyperemia, suggesting a novel role for iNOS in gastric mucosal protection. PMID- 12646422 TI - Serotonin availability is increased in mucosa of guinea pigs with TNBS-induced colitis. AB - 5-HT released from enterochromaffin cells acts on enteric nerves to initiate motor reflexes. 5-HT's actions are terminated by a serotonin reuptake transporter (SERT). In this study, we tested the hypothesis that inflammation leads to altered mucosal 5-HT signaling. Colitis was induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS), and experiments were conducted on day 6. 5-HT content, number of 5-HT-immunoreactive cells, and the proportion of epithelial cells that were 5-HT-immunoreactive increased twofold in colitis. The amount of 5-HT released under basal and stimulated conditions was significantly increased in colitis. SERT inhibition increased the 5-HT concentration in media bathing stimulated control tissue to a level comparable to that of the stimulated colitis tissue. mRNA encoding SERT and SERT immunoreactivity were reduced during inflammation. Slower propulsion and reduced sensitivity to 5-HT-receptor antagonism were observed in colitis. These data suggest that colitis alters 5-HT signaling by increasing 5-HT availability while decreasing 5-HT reuptake. Altered 5-HT availability may contribute to the dysmotility of inflammatory bowel disease, possibly due to desensitization of 5-HT receptors. PMID- 12646423 TI - Helical CT: the revolution in imaging. PMID- 12646425 TI - Review. The resistive index in renal Doppler sonography: where do we stand? PMID- 12646426 TI - Developing a prediction rule to assess hepatic malignancy in patients with cirrhosis. AB - OBJECTIVE: The objectives of our study were to identify independent clinical, demographic, and MR imaging correlates of malignancy in patients with cirrhosis and to develop a predictive model based on identified correlates of malignancy. MATERIALS AND METHODS: Sixty examinations of 58 patients with biopsy proof of lesions suggestive of hepatocellular carcinoma on MR imaging were retrospectively reviewed. The signal intensity of the lesion on T2-weighted imaging and dynamic gadolinium-enhanced imaging, the size of the lesion, and the number of suspicious lesions were recorded; in addition, patient age and sex, alpha-fetoprotein level, and hepatitis C viral genotype were noted. The association between malignancy and each predictor variable was evaluated using the chi-square test or the two-group t test. The final logistic regression model included the variables that were shown to have a significant association with malignancy and the clinically relevant predictors. We used the adjusted odds ratios to measure the strength of each association. The discriminant ability of the model for detecting hepatic malignancy was assessed using receiver operating characteristic curve analysis. RESULTS: The prevalence of hepatic malignancy in our study population was 64%. The area under the receiver operating characteristic curve for the logistic regression model was 0.82. Venous washout (odds ratio = 9.2), alpha-fetoprotein level (odds ratio = 3.2), and number of lesions (odds ratio = 1.5) were significant predictors for malignancy (p < 0.05). When arterial enhancement and venous washout were either both present or both absent, alpha-fetoprotein level contributed little to the prediction of malignancy. CONCLUSION: The MR characteristics of hepatic lesions during the dynamic venous phase in conjunction with the serum alpha-fetoprotein level and number of lesions are predictors of hepatic malignancy. The use of these predictors can facilitate explicit estimation of malignancy in individuals with underlying cirrhosis, potentially improving clinical decision-making. PMID- 12646427 TI - MR imaging of the ipsilateral breast in women with percutaneously proven breast cancer. AB - OBJECTIVE: The purpose of this study was to review MR imaging findings in the ipsilateral breast in women with percutaneously proven breast cancer. MATERIALS AND METHODS: Retrospective review was performed of records of 70 consecutive women with percutaneously proven unilateral breast cancer who were considered candidates for breast conservation surgery and who had preoperative MR imaging of the ipsilateral breast. MR images and medical records were reviewed. RESULTS: MR imaging identified mammographically and clinically occult cancer other than the index lesion in the ipsilateral breast in 19 women (27%), including infiltrating cancer in 11 women (16%) and ductal carcinoma in situ in eight women (11%). These additional sites of cancer were in the same quadrant as the index cancer in 14 women (20%), in a different quadrant in three women (4%), and in both the same and different quadrants in two women (3%). Additional sites of cancer were more likely in women with, rather than in those without, a family history of breast cancer (42% vs 14%, p < 0.02) and in women whose index cancer was infiltrating lobular rather than other histologies (55% vs 22%, p < 0.06). In 17 women (24%), MR imaging detected ipsilateral lesions that were benign. Changes due to prior percutaneous biopsy were infrequently observed on MR images and included a clip in 12 women (17%) and a small hematoma in two women (3%). CONCLUSION: MR imaging identified additional sites of ipsilateral cancer in 27% of women with percutaneously proven breast cancer. The yield was highest in women with a family history of breast cancer or infiltrating lobular histology in the index cancer. Change after biopsy was infrequent and did not interfere with the MR imaging interpretation. PMID- 12646428 TI - CT versus radiography for initial evaluation of cervical spine trauma: what is the standard of care? PMID- 12646429 TI - Fundamentals of clinical research for radiologists. Introduction to probability theory and sampling distributions. PMID- 12646430 TI - Opinion. Report of the 2002 Intersociety Commission meeting: Radiology 2002- today's research Is tomorrow's practice. PMID- 12646431 TI - Using a saline chaser to decrease contrast media in abdominal CT. AB - OBJECTIVE: The purpose of this study was to compare hepatic tumor conspicuity on CT after injection of either 150 mL of contrast material or 100 mL of contrast material plus a 50-mL saline chaser. SUBJECTS AND METHODS: We evaluated 86 hypoattenuating liver metastases in 26 patients. Patients underwent CT in two sessions separated by a mean of 85 days: one time with 150 mL of contrast material and the other time with 100 mL of contrast material followed by a 50-mL saline chaser. The order of the sessions was randomized. Contrast material was administered via power injector and matched for injection rate and delay time. Attenuation values were obtained from normal liver tissue and metastases and from the spleen, kidney, aorta, and inferior vena cava. RESULTS: The 150 mL dose of contrast material caused slightly greater liver and tumor attenuation than 100 mL of contrast material with a chaser (mean hepatic attenuation, 95.6 vs 89.8 H, respectively; p < 0.03, paired t test; mean tumor attenuation, 53.2 vs 49.1 H, respectively; r = 0.71, p = 0.09). The difference in conspicuity of liver lesions was slightly greater with 150 mL than with 100 mL with a chaser (46.8 H vs 44.2 H; r = 0.46, p = 0.08, paired t test), but was of doubtful clinical significance (2.6 H). Kidney, spleen, and vascular structures enhanced more with 150 mL than with 100 mL and a chaser. CONCLUSION: Using 100 mL of contrast material and a saline chaser did not result in a meaningful difference in liver parenchyma attenuation or lesion conspicuity compared with using 150 mL of contrast medium alone. Routine use of a chaser for abdominal CT may yield cost savings and a decreased risk of contrast nephropathy. PMID- 12646432 TI - Sydney Breast Imaging Accuracy Study: Comparative sensitivity and specificity of mammography and sonography in young women with symptoms. AB - OBJECTIVE: We examined the age-specific sensitivity and specificity of mammography and sonography in symptomatic women to determine the age below which sonography may be the more accurate imaging test, which may guide the choice of initial breast imaging examination based on the woman's age. MATERIALS AND METHODS: Four hundred eighty subjects were sampled from all women consecutively attending a symptomatic breast clinic between 1994 and 1996 and ranging in age from 25 to 55 years. We included all 240 women shown to have breast cancer (thus avoiding selection bias) and 240 age-matched women shown not to have cancer. Mammograms and sonograms were prospectively interpreted independently and without knowledge of age by two radiologists in a blinded manner, with a third radiologist arbitrating disagreements. Sensitivity and specificity of each imaging test in relation to age were examined using logistic regression modeling, and accuracy was compared using the chi-square test for paired proportions. RESULTS: Sensitivity and specificity of each test were not linearly associated with age; however, the sensitivity of mammography increased substantially in women older than 50 years. Sonographic sensitivity of 81.7% was not significantly greater than mammographic sensitivity of 75.8% (chi(2)(1) = 2.06, p = 0.15). However, in women 45 years old or younger, the sensitivity of sonography was 13.2% (95% confidence interval, 2.1-24.3%) greater than that of mammography. The specificity of both tests was approximately 88.0%. CONCLUSION: These data show that sonography is the more accurate imaging test in women 45 years old or younger who present with breast symptoms and may be an appropriate initial imaging examination. PMID- 12646433 TI - Sonographic detection and sonographically guided biopsy of breast microcalcifications. AB - OBJECTIVE: The purpose of this study was to evaluate the ability of sonography to depict and guide biopsies of mammographically suspicious microcalcifications and to reveal the mammographic features and histologic outcomes of lesions amenable to sonographically guided biopsy. SUBJECTS AND METHODS: . Suspicious clusters of microcalcifications without other mammographic abnormalities were evaluated on sonography before biopsy and divided into two groups: those with and those without microcalcifications seen on sonography. Sonographically detected lesions underwent sonographically guided biopsy; lesions not seen on sonography underwent mammographically guided biopsy. Imaging features and histologies were correlated, and the positive predictive value of sonography was determined. RESULTS: Of 111 lesions (105 patients), 26 lesions (23%) were identified and underwent sonographically guided biopsy; 85 lesions (77%) were not identified sonographically. The diameters of microcalcification clusters in the sonographically identified group were significantly larger (p = 0.0005) and contained larger numbers of microcalcification particles (p = 0.038) compared with clusters not identified sonographically. Sonographically identified lesions were seen as masses (77%) or dilated ducts (23%) with echogenic foci. Sonographically identified lesions were more likely to be malignant than those not seen on sonography (69% vs 21%, respectively; p < 0.00002). Of 38 malignant lesions, those visible on sonography were more likely to be invasive than those not seen on sonography (72% vs 28%, respectively; p = 0.018). In malignant lesions undergoing core biopsy and surgical excision, the extent of disease was underestimated less with sonographically guided biopsy (7%, 1/15) than with stereotactic biopsy (33%, 5/15). CONCLUSION: Suspicious microcalcifications are seen infrequently on sonography (23%) but, when detected, can be successfully biopsied with sonographic guidance and more frequently are malignant and represent invasive cancer than those seen on mammography alone. PMID- 12646434 TI - Original report. Mucocele-like tumors of the breast: mammographic and sonographic appearances. AB - OBJECTIVE: The purpose of our study was to describe the mammographic and sonographic appearances of the mucocele-like tumor of the breast. CONCLUSION: The mucocele-like tumor is a rare lesion of the breast, the benign form of which has a nonspecific mammographic appearance. The tumor can present as indeterminate microcalcifications or as a nodule, often containing calcifications. The sonographic findings are of multiple well-defined hypoechoic oval or tubular structures with low-level internal echoes resembling complex cysts. Mucocele-like tumors of the breast can be associated with atypical hyperplasia or carcinoma. If a mucocele-like tumor is diagnosed at core needle biopsy, complete surgical excision is recommended, with careful evaluation of the entire specimen to exclude the presence of atypia or carcinoma. PMID- 12646435 TI - Small solitary pulmonary nodules (< or =1 cm) detected at population-based CT screening for lung cancer: Reliable high-resolution CT features of benign lesions. AB - OBJECTIVE: We assessed thin-section CT features specific to benignity in solitary pulmonary nodules of 1 cm or smaller that were detected at population-based CT screening for lung cancer. MATERIALS AND METHODS: Two reviewers independently made qualitative (presence or absence of lobulation, spiculation, air bronchogram, cavity, satellite lesions, pleural tag, concave margins, polygonal shape, and peripheral subpleural lesion) and quantitative (lesion size, percentage of ground-glass opacity areas, and two- and three-dimensional ratios of lesion) assessments in CT images of 72 nodules (25 lung cancers, seven atypical adenomatous hyperplasias, and 40 benign lesions). Optimal criteria specific to benignity were studied. RESULTS: The prevalence of polygonal shape (p = 0.005 and p = 0.019, reviewer 1 and reviewer 2), peripheral subpleural lesion (p = 0.011 and p = 0.033), a predominantly solid lesion (p < 0.001 and p < 0.001), and three-dimensional ratios (p < 0.001 and p < 0.001) were greater in benign lesions than in malignancies. The prevalence of a predominantly solid lesion (p < 0.001 and p < 0.001) was greater in benign lesions than in atypical adenomatous hyperplasias, and the prevalence of a peripheral subpleural lesion (p = 0.004 and p = 0.012) was greater in atypical adenomatous hyperplasias than in malignancies. Using a single CT feature, polygonal shape and a three-dimensional ratio of greater than 1.78 showed 100% specificity for both reviewers. Among all combinations of CT findings specific to benignity, a combined criterion of a predominantly solid lesion and peripheral subpleural lesion or polygonal shape or the three-dimensional ratio attained the highest sensitivity (63% and 60%) for both reviewers. CONCLUSION: A combined criterion of CT features was optimal for predicting benign pulmonary lesions. PMID- 12646436 TI - Widespread ground-glass opacity of the lung in consecutive patients undergoing CT: Does lobular distribution assist diagnosis? AB - OBJECTIVE: Our purpose was to establish the most frequent diagnoses associated with widespread ground-glass opacity on CT and to determine if the distribution of ground-glass opacity at a lobular level affects the likelihood of a given diagnosis. MATERIALS AND METHODS: A retrospective search of our department's CT database from January 1998 through December 2000 revealed 153 cases of ground glass opacity. Patients evaluated using bronchoscopy or open biopsy within 30 days (mean, 6.9 days) of imaging with ground-glass opacity as the predominant high-resolution CT finding were selected. There were 21 men and 16 women with a mean age of 51.4 years. Two chest radiologists, unaware of the clinical diagnoses, independently recorded lobular distributions. Consensus interpretation was used for discrepancies. Primary distributions were recorded as lobular (geographic ground-glass opacity marginated by septal anatomy), centrilobular (ground-glass opacity related to bronchovascular anatomy), or random. RESULTS: Infectious and other histologic diagnoses fell into four diagnostic groups: atypical pneumonia, chronic infiltrative interstitial disease, acute air-space filling, and drug toxicity. Ground-glass opacity was most frequently associated with acute atypical pneumonia (n = 12, 32%), chronic infiltrative disease (n = 10, 27%), acute air-space filling (n = 6, 16%), and drug toxicity (n = 4, 11%). In five patients, a definitive diagnosis was not established. Ground-glass opacity was most commonly randomly distributed (n = 16, 43%), followed by lobular (n = 15, 41%) and centrilobular (n = 6, 16%) distributions. Distribution did not correlate with diagnostic group. CONCLUSION: In unselected cases of ground-glass opacity evaluated at a tertiary institution, atypical infection and chronic infiltrative interstitial disease accounted for 59% of diagnoses. Distribution at a lobular level did not differentiate underlying causes. PMID- 12646437 TI - Case report. Pyothorax-associated lymphoma: Diagnosis at percutaneous core biopsy with CT guidance. PMID- 12646438 TI - Radiologic-pathologic conferences of the University of Texas M. D. Anderson Cancer Center: Extraskeletal osteosarcoma of the pleura. PMID- 12646439 TI - Pictorial essay. Multidetector CT evaluation of congenital heart disease in pediatric and adult patients. PMID- 12646440 TI - Original report. Late myocardial enhancement in hypertrophic cardiomyopathy with contrast-enhanced MR imaging. AB - OBJECTIVE: Contrast-enhanced MR imaging has great clinical value as a means of characterizing myocardial tissue in patients with ischemic heart disease. The purpose of our study was to evaluate use of this technique in the assessment of patients with hypertrophic cardiomyopathy. CONCLUSION: On late-enhancement MR imaging, the various types or patterns of enhancement found in patients with hypertrophic cardiomyopathy are related to differences in morphology and regional function. Enhancement in hypertrophied areas likely reflects the presence of abundant connective tissue, foci of myocardial necrosis, or a combination of both. PMID- 12646441 TI - Trauma cases from Harborview Medical Center. Doll's head "bezoar": complete craniocervical dislocation causing bowel obstruction. PMID- 12646442 TI - Videofluoroscopy of swallowing abnormalities in 22 symptomatic patients after cardiovascular surgery. AB - OBJECTIVE: The aim of this study was to evaluate the types of swallowing abnormalities that occur in symptomatic patients who have undergone cardiovascular surgery. MATERIALS AND METHODS: From 1994 to 2001, 22 patients (17 males and five females; age range, 4-89 years; mean age, 64 years) who had swallowing abnormalities after cardiovascular surgery were referred for a videofluoroscopic swallowing study. Each study was analyzed for functional abnormalities of the tongue, soft palate, epiglottis, hyoid and larynx, pharynx, upper esophageal sphincter, and esophagus. Also, the performance of transesophageal echocardiography, long-term intubation, or both was noted. RESULTS: Swallowing abnormalities were present in 18 patients (81.8%) (range, one to eight functional abnormalities; mean, 3.9 functional abnormalities). The distribution of abnormalities across the functional units statistically significantly deviated (chi(2) = 14.4; df = 6; p = 0.025) from uniform distribution, with abnormalities most commonly involving the hyoid and larynx (13 patients [59.1%]) and the pharynx (10 patients [45.5%]). Aspiration was found in 13 patients (59.1%) (predeglutitive, n = 1; intradeglutitive, n = 4; postdeglutitive, n = 3; and mixed, n = 5). In the 14 patients (63.6%) who underwent transesophageal echocardiography, long-term intubation, or both, we frequently found incomplete tilting of the epiglottis, pharyngeal weakness, and postdeglutitive aspiration. CONCLUSION: Most patients with swallowing problems after cardiovascular surgery present with multiple abnormalities that most commonly affect the hyoid and larynx and the pharynx and result predominantly in intra- or postdeglutitive aspiration. The performance of transesophageal echocardiography and long-term intubation may influence the types of swallowing abnormalities. PMID- 12646443 TI - Using a phantom to compare MR techniques for determining the ratio of intraabdominal to subcutaneous adipose tissue. AB - OBJECTIVE: Patients who have a greater distribution of intraabdominal adipose tissue as compared with subcutaneous adipose tissue and an increased ratio of intraabdominal adipose tissue to subcutaneous adipose tissue are at greater risk for developing cardiovascular disease and type 2 diabetes mellitus. In previous MR investigations, researchers have used conventional T1-weighted spin-echo images to determine the ratio of intraabdominal adipose tissue to subcutaneous adipose tissue. However, no investigation, to our knowledge, has been performed to determine the accuracy of using different MR sequences to estimate adipose distribution. The purpose of our investigation was to compare MR imaging and segmentation techniques in calculating the ratio of intraabdominal to subcutaneous adipose tissue using an adiposity phantom. MATERIALS AND METHODS: A phantom was created to simulate the distribution of subcutaneous and intraabdominal fat (with known volumes). Axial MR images were obtained twice through the phantom using a 5-mm slice thickness and zero gap for the following T1-weighted sequences: spin-echo, fast Dixon, and three-dimensional (3D) spoiled gradient-echo. An in-house computer software program was then used to segment the volumes of fat and calculate the volume of intraabdominal adipose tissue and subcutaneous adipose tissue and the ratio of intraabdominal to subcutaneous adipose tissue. Each imaging data set was segmented three times, so six sets of data were yielded for each imaging technique. The percentage predicted of the true volume was calculated for each MR imaging technique for each fat variable. The mean percentages for each variable were then compared using one-factor analysis of variance to determine whether differences exist among the three MR techniques. RESULTS: The three MR imaging techniques had statistically significant different means for the predicted true volume of two variables: volume of subcutaneous adipose tissue (p < 0.001) and volume of intraabdominal adipose tissue (p = 0.0426). Estimates based on fast Dixon images were closest to the true volumes for all the variables. All MR imaging techniques performed similarly in estimating the ratio of intraabdominal adipose tissue to subcutaneous adipose tissue (p = 0.9117). The acquisition time for the 3D spoiled gradient-echo images was 10-22 times faster than for the other sequences. CONCLUSION: Conventional T1-weighted spin-echo MR imaging, the current sequence used in practice for measuring visceral adiposity, may not be the optimal MR sequence for this purpose. We found that the T1-weighted fast Dixon sequence was the most accurate at estimating all fat volumes. The T1-weighted 3D spoiled gradient-echo sequence generated similar ratios of intraabdominal to subcutaneous adipose tissue in a fraction of the acquisition time. PMID- 12646444 TI - Pictorial essay. Radiology of ileal pouch-anal anastomosis surgery. PMID- 12646445 TI - Case report. CT diagnosis of volvulus of the descending colon with persistent mesocolon. PMID- 12646446 TI - Helical CT screening for hepatocellular carcinoma in patients with cirrhosis: frequency and causes of false-positive interpretation. AB - OBJECTIVE: The purpose of our study was to determine the specificity of helical CT for depiction of hepatocellular carcinoma in a population of patients with cirrhosis. SUBJECTS AND METHODS: Single-detector helical CT screening was undertaken in 1329 patients with cirrhosis who were referred for transplantation. The patients underwent one or more helical CT examinations over 30 months and were followed up for an additional 19 months or until transplantation. We predominantly used unenhanced and biphasic contrast-enhanced techniques with infusions of 2.5-5.0 mL/sec. Four hundred thirty patients underwent transplantation within this period. Liver specimens were sectioned at 1-cm intervals, with direct comparison of imaging and pathologic findings and histologic confirmations of all lesions. Prospective preoperative helical CT reports were used for the primary data analysis. A retrospective unblinded review was undertaken to determine characteristics of false-positive lesions diagnosed as hepatocellular carcinoma. RESULTS: Thirty-five patients (8%) had false positive diagnoses for hepatocellular carcinoma based on helical CT. Twenty of these patients (5%) showed hypoattenuating lesions seen during one of the three helical CT examination phases. Fifteen patients (3%) had hyperattenuating lesions seen during the arterial phase. Among the 15 hyperattenuating lesions, CT revealed the causes to be transient benign hepatic enhancement (n = 3), hemangiomas (n = 2), fibrosis (n = 2), peliosis (n = 1), volume averaging (n = 1), low-grade dysplastic nodule (n = 1), or undetermined (n = 5). Of the 20 hypoattenuating lesions, the causes were shown to be fibrosis (n = 8), focal fat (n = 4), infarcted regenerative nodules (n = 2), regenerative nodules (n = 1), fluid trapped at the dome of the liver (n = 1), hemangioma (n = 1), or undetermined (n = 3). Follow-up helical CT in 13 (72%) of 18 patients allowed a change in the diagnosis of hepatocellular carcinoma to a finding of no cancer present. CONCLUSION: Helical CT screening for hepatocellular carcinoma in patients with cirrhosis has a substantial false-positive detection rate. Although most of lesions were hypoattenuating, a few hyperenhancing arterial phase lesions were proven not to be hepatocellular carcinoma. An awareness of imaging characteristics and follow-up imaging can help radiologists avoid a mistaken diagnosis in many patients. PMID- 12646447 TI - Pictorial essay. Hepatocellular carcinoma: imaging and imaging-guided intervention. PMID- 12646448 TI - Nondysplastic nodules that are hyperintense on T1-weighted gradient-echo MR imaging: frequency in cirrhotic patients undergoing transplantation. AB - OBJECTIVE: Our objective was to determine the frequency and MR imaging findings of nondysplastic nodules that are hyperintense on T1-weighted gradient-echo imaging in patients with cirrhosis who undergo liver transplantation. MATERIALS AND METHODS: Two observers retrospectively evaluated in-phase (4-5 msec), opposed phase gradient-echo (2.0-2.4 msec), and turbo short tau inversion recovery (STIR) MR images in 68 patients with cirrhosis--but without dysplastic nodules or hepatocellular carcinoma--who underwent MR imaging at 1.5 T within 150 days before liver transplantation. The size, number, signal characteristics, and arterial enhancement pattern of nodules that appear hyperintense on T1-weighted gradient-echo images were evaluated as well as the presence or absence of signal loss on opposed-phase imaging. These imaging findings were correlated with pathologic findings of whole explanted livers. RESULTS: Eleven (16%) of 68 patients had at least one nondysplastic nodule that was hyperintense on T1 weighted MR imaging. Three patients had diffuse nondysplastic hyperintense nodules (>10 nodules) measuring less than 0.5 cm, and the remaining eight patients had 22 nondysplastic hyperintense nodules ranging in size from 0.5 to 2.5 cm (mean, 1.2 cm), of which 13 were isointense and nine were hypointense on turbo STIR images. No lesion lost signal on opposed-phase imaging or enhanced during the hepatic arterial phase. CONCLUSION: In cirrhotic patients undergoing liver transplantation, nondysplastic nodules that are hyperintense are common findings on T1-weighted gradient-echo MR imaging and do not lose signal intensity on opposed-phase imaging or enhance during the hepatic arterial phase. These nodules may be indistinguishable from dysplastic nodules. PMID- 12646449 TI - Case report. Intrabiliary colonic metastasis mimicking primary biliary neoplasia. PMID- 12646450 TI - Case report. CT, MR cholangiopancreatography, and endoscopy findings in Bouveret's syndrome. PMID- 12646452 TI - The urethra and its supporting structures in women with stress urinary incontinence: MR imaging using an endovaginal coil. AB - OBJECTIVE: The objective of this study was to evaluate the urethra and its supporting structures in patients with stress urinary incontinence using MR imaging with an endovaginal coil. SUBJECTS AND METHODS: We reviewed MR images obtained using an endovaginal coil in 63 patients with stress urinary incontinence and in 16 continent women. We compared the two groups for the thickness of the striated muscle, smooth muscle, and mucosa-submucosa of the urethra; degree of asymmetry of the puborectalis muscle; frequency of distortion in the periurethral, paraurethral, and pubourethral ligaments; degree of the vesicourethral angle; and dimension of the retropubic space. Using the status of the urethra and its supporting structures as our basis, we scored the risk of stress urinary incontinence for each woman on a scale of 0-5. RESULTS: The striated muscle layer of the urethra was thinner in the group with stress urinary incontinence (mean +/- SD, 1.9 +/- 0.5 mm) than that in the continent group (2.6 +/- 0.4 mm) (p < 0.001). A high degree of asymmetry of puborectalis muscle (>1.5) was more frequent in the group with stress urinary incontinence (29%) than in the continent group (0%) (p = 0.015). Supporting ligaments were more frequently distorted in the incontinent group than in the continent group. Distorted periurethral ligaments were found in 56% of the patients with stress urinary incontinence versus 13% of the women who were continent; distorted paraurethral ligaments were found in 83% of the patients with stress urinary incontinence versus 19% of the women who were continent; and distorted pubourethral ligaments were found in 54% of the patients with stress urinary incontinence versus 19% of the women who were continent (p < 0.05). The group with stress urinary incontinence had a greater vesicourethral angle (148 degrees vs 125 degrees ) and larger retropubic space (7.5 vs 5.1 mm) than did the women who were continent (p < 0.05). The score for the risk of stress urinary incontinence was higher in the group with stress urinary incontinence (3.3 +/- 1.4) than in the women who were continent (1.0 +/- 1.2) (p < 0.001). CONCLUSION: MR imaging with an endovaginal coil revealed significant morphologic alterations of the urethra and supporting structures in patients with stress urinary incontinence. PMID- 12646453 TI - Imaging in oncology from the University of Texas M. D. Anderson Cancer Center. Imaging in the diagnosis, staging, and follow-up of cancer of the urinary bladder. PMID- 12646454 TI - Early experience with percutaneous radiofrequency ablation of small solid renal masses. AB - OBJECTIVE: Incidental small renal-cell masses are often seen in elderly patients with significant comorbidity who are unfit to undergo major surgery. This study was conducted to determine the safety and efficacy of percutaneous imaging-guided radiofrequency ablation in the management of small solid renal masses, almost all of which are renal cell cancers. CONCLUSION: Early experience suggests that radiofrequency ablation is a safe, well-tolerated, and minimally invasive therapy for patients with solid renal masses. In the era of nephron-sparing surgery, radiofrequency ablation may have a role in the management of small problematic renal masses. PMID- 12646455 TI - Original report. Pulse-spray thrombolysis of thrombosed hemodialysis grafts with tissue plasminogen activator. AB - OBJECTIVE: The purpose of this study was to evaluate pulse-spray pharmacomechanical thrombolysis with the use of tissue plasminogen activator in the recanalization of thrombosed hemodialysis access grafts. CONCLUSION: Pulse spray pharmacomechanical thrombolysis with tissue plasminogen activator is an effective method for percutaneous recanalization of thrombosed hemodialysis access grafts with results similar to other percutaneous techniques. PMID- 12646456 TI - Technical innovation. Using a covered stent (wallgraft) to treat pseudoaneurysms of dialysis grafts and fistulas. PMID- 12646457 TI - Technical innovation. Using a cutting balloon to treat resistant high-grade dialysis graft stenosis. PMID- 12646458 TI - Technical innovation. Combined treatment of a spinal metastasis with radiofrequency heat ablation and vertebroplasty. PMID- 12646459 TI - Interventional radiology case conferences Massachusetts General Hospital. Continuous high-output drainage of hepatic abscess 3 months after radiofrequency ablation of hepatocellular carcinoma. PMID- 12646460 TI - Multidetector CT angiography of the aortoiliac system and lower extremities: a prospective comparison with digital subtraction angiography. AB - OBJECTIVE: Our objective was to determine whether multidetector CT (MDCT) angiography is an accurate and reliable method of revealing atheroocclusive disease of the aortoiliac system and the lower extremities compared with digital subtraction angiography (DSA). SUBJECTS AND METHODS: Forty-one patients with ischemic legs underwent both MDCT angiography and DSA of the aortoiliac system and the legs. The arterial supply of the legs was divided into 35 segments. Three independent observers rated each segment according to the maximal degree of arterial stenosis. Consensus interpretation was used to calculate the sensitivity and specificity of MDCT angiography in showing arterial occlusions and stenoses of at least 75%. Intertechnique agreement was measured for each anatomic segment, and interobserver agreement was calculated for both techniques. Agreement was quantified using the kappa statistic. RESULTS: The sensitivity and specificity of MDCT angiography for depicting arterial occlusions and stenoses of at least 75% were 88.6% and 97.7%, and 92.2% and 96.8%, respectively. Substantial intertechnique agreement (kappa > 0.4) was present in 102 (97.1%) of 105 arterial segments. Substantial interobserver agreement was present in 104 (99.0%) of 105 comparisons for both MDCT angiography and DSA with an average kappa value of 0.84 for CT and 0.78 for DSA. MDCT angiography showed more patent segments than DSA (1192 vs 1091). All nine segments seen on DSA and not seen on MDCT angiography were in the calves. Of 110 segments seen on MDCT angiography and not seen on DSA, 100 (90.9%) were in the calves. CONCLUSION: MDCT angiography was accurate in showing arterial atheroocclusive disease with reliability similar to DSA. MDCT angiography showed more vascular segments than DSA, particularly within calf vessels. PMID- 12646461 TI - Technical innovation. Optimization of multidetector CT venography performed with elastic stockings on patients' lower extremities: A preliminary study of nonthrombosed veins. PMID- 12646462 TI - MR imaging, MR arthrography, and specimen correlation of the posterolateral corner of the knee: an anatomic study. AB - OBJECTIVE: We sought to evaluate the anatomy of the posterolateral aspect of the knee with anatomic dissection, MR imaging, MR arthrography, and sectional anatomy. MATERIALS AND METHODS: We assessed the posterolateral corner of the knee during dissection of one gross anatomic specimen. MR imaging and MR arthrography were performed in seven additional knee specimens. T1-weighted spin-echo MR images were obtained in the standard imaging planes as well as in the coronal oblique plane. The specimens underwent T1-weighted spin-echo MR imaging after administration of intraarticular contrast material and were sectioned into planes corresponding to those of the MR images. RESULTS: At anatomic dissection, the following posterolateral structures were identified: the arcuate ligament (medial and lateral limbs), fabellofibular ligament, popliteofibular ligament, popliteus tendon and its two posterior attachments to the lateral meniscus, fibular collateral ligament, direct and anterior arms of the tendon of the long head of the biceps femoris muscle, and direct and anterior arms of the tendon of the short head of the biceps femoris muscle. Correlation of MR imaging and anatomic findings showed that the popliteofibular ligament and oblique popliteal ligament were found in 57% and 100% of specimens, respectively. At least one of the two limbs of the arcuate ligament was identified in 71% of specimens. The fabellofibular ligament was not identified on MR images in any of the specimens. The anteroinferior and posterosuperior popliteomeniscal fascicles were identified in all specimens. CONCLUSION: The posterolateral corner of the knee comprises complex and variable anatomic structures. Recognition of these variations is important in the assessment of MR images of the knee. PMID- 12646463 TI - Pictorial essay. MR imaging appearance and classification of acromioclavicular joint injury. PMID- 12646464 TI - Comparison of imaging-guided and non-imaging-guided quantitative sonography of the calcaneus with dual X-ray absorptiometry of the spine and femur. AB - OBJECTIVE: The purpose of our study was to evaluate the diagnostic agreement between imaging-guided and non-imaging-guided quantitative sonography of the calcaneus and dual X-ray absorptiometry of the spine and femur to show osteoporosis. SUBJECTS AND METHODS: In 113 patients (73 women, 59 +/- 14 years old; 40 men, 48 +/- 16 years old), dual X-ray absorptiometry of the lumbar spine and the proximal femur, imaging-guided quantitative sonography, and non-imaging guided quantitative sonography of the calcaneus were performed. The percentage of patients having a T-score equal to or less than a threshold of -2.5 SDs (prevalence of osteoporosis) was calculated for each imaging technique. The diagnostic agreement of the three techniques in identifying individuals with osteoporosis was assessed. RESULTS: Eleven percent of the women and 8% of the men were classified as osteoporotic by imaging-guided quantitative sonography, and 38% of the women and 25% of the men were so classified by non-imaging-guided quantitative sonography. At dual X-ray absorptiometry of the spine, 44% of the women and 38% of the men were classified as osteoporotic, and, at different femoral regions, 19-60% of the women and 8-38% of the men were so classified. Kappa analysis for both quantitative sonography techniques was not significant. Kappa analysis for both quantitative sonography techniques and dual X-ray absorptiometry showed diagnostic agreement to be generally poor. CONCLUSION: No advantage in diagnostic accuracy could be found for imaging-guided quantitative sonography. The considerable diagnostic disagreement between both quantitative sonography techniques and dual X-ray absorptiometry could be confusing in daily clinical practice. PMID- 12646465 TI - Original report. Sonographic evaluation of shoulder arthroplasty. AB - OBJECTIVE: The objective of our study was to review our experience using sonography to evaluate the rotator cuff after arthroplasty. Diagnostic reliability, with respect to surgical findings, as well as subjective assessment of the examinations, including the usefulness of applications such as extended field-of-view imaging, photopic imaging, and tissue harmonic imaging, were reviewed. CONCLUSION: We found sonography to be a useful method of imaging the rotator cuff after arthroplasty. Extended field-of-view imaging and tissue harmonic imaging aided in diagnosis by improving visualization of regional anatomic landmarks and increasing conspicuity of small tendon tears. PMID- 12646466 TI - Technical innovation. Dynamic sonography of the forefoot: The sonographic Mulder sign. PMID- 12646468 TI - Comparison of oral pentobarbital sodium (nembutal) and oral chloral hydrate for sedation of infants during radiologic imaging: preliminary results. AB - OBJECTIVE: The purpose of this study was to compare the safety and efficacy of oral cherry-flavored pentobarbital sodium (Nembutal) and oral chloral hydrate to sedate infants undergoing radiologic imaging. SUBJECTS AND METHODS: We prospectively recorded data for all infants sedated with oral cherry-flavored pentobarbital sodium and oral chloral hydrate for imaging examinations between January 1997 and August 1999. The parameters recorded were each patient's age, weight, and American Society of Anesthesiologists classification; the time required to sedate; the total length of sedation time; the time required to discharge from the recovery room; and adverse events. The two-sample Student's t test and Fisher's exact test were used for statistical analysis. RESULTS: Oral pentobarbital sodium was administered to 317 infants. These infants had a mean age +/- SD of 6.9 +/- 3.1 months and a mean weight of 7.8 +/- 4.8 kg; they received a median dose of 4 mg/kg of body weight. Oral chloral hydrate was administered to 358 infants. These infants had a mean age of 5.9 +/- 3.3 months and a mean weight of 7.3 +/- 4.9 kg; they received a median dose of 50 mg/kg of body weight. The mean time required to sedate was 19 +/- 14 min for infants receiving oral pentobarbital sodium and 16 +/- 11 min for infants receiving oral chloral hydrate (p = 0.02); the mean time required to discharge was 100 +/- 35 min for infants in the oral pentobarbital sodium group and 103 +/- 36 min for infants in the oral chloral hydrate group (p = 0.31); the mean length of sedation was 81 +/- 34 min for the oral pentobarbital sodium group and 86 +/- 36 min for the oral chloral hydrate group (p = 0.07); and median American Society of Anesthesiologists classification for both groups was P1. Oral pentobarbital sodium was inadequate for sedation in one patient (0.3%) and chloral hydrate was inadequate for sedation in another (0.3%) (p = 1.00). Adverse events were recorded for five patients (1.6%) in the oral pentobarbital sodium group and for six patients (1.7%) in the chloral hydrate group (p = 0.99). CONCLUSION: Oral pentobarbital sodium is as safe and efficacious as oral chloral hydrate for sedating infants. PMID- 12646469 TI - CT findings and temporal course of persistent pulmonary interstitial emphysema in neonates: a multiinstitutional study. AB - OBJECTIVE: The purpose of this study was to evaluate the CT appearance, management, and temporal course of persistent pulmonary interstitial emphysema in neonates. MATERIALS AND METHODS: Criteria for inclusion in the study group included neonates with a history of prematurity who required ventilation for lung disease, development of hyperexpanded radiolucent lung lesions after typical radiographic findings of pulmonary interstitial emphysema, and CT documentation of lung abnormalities. Radiographs and CT scans were reviewed for the anatomic distribution, appearance, and presence of classic lines or dots within a radiolucent mass. We compared the management (surgical vs nonsurgical) and the temporal course in nonsurgical cases for patients in the United States and patients outside the United States. RESULTS: From seven institutions, we identified 17 patients who had persistent pulmonary interstitial emphysema with CT documentation. On CT, all lesions consisted of hyperexpanded cystic radiolucencies. Distribution was single-lobe in nine patients (left upper lobe, n = 5; left lower lobe, n = 3; right middle lobe, n = 1), multilobar in eight patients, and bilateral in six patients. Fourteen patients (82%) showed the characteristic line-and-dot pattern. All patients were initially treated conservatively; nine eventually underwent surgical resection with confirmation at pathology. Of 10 patients who underwent nonsurgical treatment for at least 1 year, lesions resolved in four patients, decreased in size in three asymptomatic patients, and enlarged in three patients, with eventual resection in two. Surgical resection was performed in 89% (8/9) of patients treated in the United States and in 13% (1/8) of patients treated outside the United States. CONCLUSION: Our study found that 82% of patients with persistent pulmonary interstitial emphysema had characteristic CT findings (central lines and dots surrounded by radiolucency). Although most patients in this series eventually underwent surgical resection, initial treatment can be conservative. The decision to perform surgery may have reflected the local medical culture. PMID- 12646471 TI - MR evaluation of arrhythmogenic right ventricular cardiomyopathy in pediatric patients. AB - OBJECTIVE: The aim of our study was to correlate the findings of three MR imaging sequences with the clinical findings of possible arrhythmogenic right ventricular cardiomyopathy in pediatric patients. MATERIALS AND METHODS: Twenty-six consecutive pediatric patients underwent MR imaging with ECG-gated non-breath hold spin-echo T1-weighted non-fat-suppressed and fat-suppressed sequences. The MR images were evaluated for thinning or fat signal in the right ventricular wall and for enlargement or increased trabeculation of the right ventricle or right ventricular outflow tract. Cine MR imaging was used to assess wall motion abnormalities. Cardiac biopsy was performed in 17 patients. Biopsy results and other clinical findings suggesting arrhythmogenic right ventricular cardiomyopathy were tabulated. RESULTS: Two MR imaging studies were of poor quality as a result of arrhythmias, and one study was incomplete. In the 23 remaining patients, there were (mean +/- SD) 1.5 +/- 1.0 and 0.8 +/- 1.0 findings of possible arrhythmogenic right ventricular cardiomyopathy in the non-fat suppressed and the fat-suppressed sequences, respectively. Fat-compatible signal in the myocardium was detected in 16 (70%) of 23 non-fat-suppressed studies and in five (22%) of 23 fat-suppressed studies (p = 0.003). The non-fat-suppressed sequence had a higher sensitivity (75% vs 43%) and a lower specificity (38% vs 75%) for fatty infiltration than did the fat-suppressed sequence when correlated with the biopsies. The linear correlation between all MR findings and all clinical diagnostic criteria, including biopsy, was better for the combination of cine and both T1 sequences (r = 0.58) than for the non-fat-suppressed (r = 0.53) or fat-suppressed (r = 0.46) T1 sequences alone. CONCLUSION: MR imaging showed moderate correlation with the clinical criteria in the diagnosis of arrhythmogenic right ventricular cardiomyopathy. PMID- 12646472 TI - Original report. Testicular torsion in neonates and infants: sonographic features in 30 patients. AB - OBJECTIVE: The purpose of this study was to illustrate the spectrum of sonographic findings in testicular torsion in a large series of neonates and infants. These patterns and their evolution have, to our knowledge, not been described previously. CONCLUSION: The sonographic appearance of testicular torsion in neonates and infants can be divided into three types. We believe that the findings represent different stages in the evolution of testicular torsion. PMID- 12646473 TI - Pictorial essay. Neonates and umbilical venous catheters: normal appearance, anomalous positions, complications, and potential aid to diagnosis. PMID- 12646474 TI - Fetal central nervous system biometry on MR imaging. AB - OBJECTIVE: We sought to compare the biometry of the fetal head on MR imaging with sonographic measurements in fetuses with and without suspected central nervous system abnormalities. MATERIALS AND METHODS: Blinded retrospective measurements of biparietal diameter, head circumference, and cerebellar width obtained on MR imaging were assigned a gestational age on the basis of median sonographic measurements and compared with sonographic and clinical assignment of gestational age in fetuses with no central nervous system abnormalities. In fetuses with central nervous system abnormalities, the same MR measurements were compared with sonographic measurements obtained within 1 week. Single-shot fast spin-echo sequences were obtained. Pearson's product moment correlation coefficients and paired sample t tests were performed. RESULTS: In 22 fetuses with no suspected central nervous system abnormalities, significant correlation was seen in the assignment of gestational age by MR measurements and sonographic gestational age. In 25 fetuses with central nervous system abnormalities, significant correlation was also seen between biparietal diameter and head circumference measurements. The mean biparietal diameter on MR imaging was greater than on sonography in those fetuses with central nervous system abnormalities (p = 0.038). CONCLUSION: MR imaging measurements of biparietal diameter, head circumference, and cerebellar width are strongly correlated to gestational age in fetuses without central nervous system abnormalities. Significant correlation is found between MR imaging and sonographic measurements of biparietal diameter and head circumference in fetuses with central nervous system abnormalities. Larger biparietal diameter measurements were seen with MR imaging than with sonography in the abnormal group. Fetal central nervous system biometry can be performed as part of the MR imaging evaluation of the fetal central nervous system. PMID- 12646475 TI - Original report. Fetal cardiographic monitoring during 1.5-T MR imaging. AB - OBJECTIVE: The aim of this report was to determine the feasibility of fetal cardiotocography during MR imaging and the safety of 1.5-T MR imaging on the basis of fetal heart activity and fetal movements. CONCLUSION: Fetal cardiotocography is feasible during MR imaging using modified standard equipment. The use of 1.5-T MR imaging appears to be safe and to have no negative short-term effects on the heart rate or movement incidence of healthy third-trimester fetuses under our experimental conditions. PMID- 12646476 TI - Using CT to localize side and level of vocal cord paralysis. AB - OBJECTIVE: The purpose of our study was to assess the relative accuracy of imaging findings related to peripheral recurrent nerve paralysis on axial CT studies of the neck. Also assessed were imaging findings of a central vagal neuropathy. MATERIALS AND METHODS: We retrospectively identified 40 patients who had clinically diagnosed vocal cord paralysis and had undergone CT. Eight imaging signs of vocal cord paralysis were assessed, and an imaging distinction between a central or peripheral vagal neuropathy was made by evaluating asymmetric dilatation of the oropharynx with thinning of the constrictor muscles. In two patients, we studied the use of reformatted coronal images from a multidetector CT scanner. RESULTS: For unilateral vocal cord paralysis, the most sensitive imaging findings were ipsilateral pyriform sinus dilatation, medial positioning and thickening of the ipsilateral aryepiglottic fold, and ipsilateral laryngeal ventricle dilatation. In two patients, coronal reformatted images aided the diagnosis by better showing flattening of the subglottic arch. Imaging findings allowed localization of a central vagal neuropathy in four patients. CONCLUSION: Three reliable imaging findings associated with vocal cord paralysis were identified on routine axial CT studies: ipsilateral pyriform sinus dilatation, medial positioning and thickening of the ipsilateral aryepiglottic fold, and ipsilateral laryngeal ventricle dilatation. Coronal reformatted images of the larynx may be helpful, but they are not necessary in 95% of patients. Ipsilateral pharyngeal constrictor muscle atrophy is a helpful imaging finding to localize a more central vagal neuropathy. Our findings can aid radiologists in identifying peripheral and central vagal neuropathy in patients who present for CT of the neck who have a normal voice and are without a history suggestive of a vagal problem. PMID- 12646477 TI - Recurrent pleomorphic adenoma of the parotid gland in pediatric and adult patients: value of multiple lesions as a diagnostic indicator. AB - OBJECTIVE: Recurrent pleomorphic adenoma of the parotid gland is a significant problem. Rates have been as high as 40% in some series of patients who have undergone surgery for the primary lesion. In the imaging literature, anecdotal case reports show multiple lesions in recurrent pleomorphic adenoma. Our aim was to analyze the imaging of a series of patients to determine the reliability of multiple lesions as a tool in diagnosing recurrent disease. MATERIALS AND METHODS: Medical records of the patients with recurrent pleomorphic adenoma of the parotid gland referred to our institution were retrospectively reviewed. Before the second surgery, MR imaging had been performed in 15 patients. We retrospectively reevaluated and scored the MR imaging studies with particular attention paid to the location and number of the lesions and the remaining parotid gland tissue. RESULTS: On the basis of imaging findings, eight patients underwent enucleation, superficial parotidectomy had been performed in four patients, and three patients underwent total parotidectomy. For our group, the lesions were multiple in 73.3% of patients. CONCLUSION: To our knowledge, we present the first large series of imaging studies in recurrent pleomorphic adenoma of the parotid gland. Our findings show that recurrent pleomorphic adenomas are most likely to be multiple. Such multiplicity of lesions is a reliable diagnostic indicator of recurrent disease. PMID- 12646478 TI - To excise or to sample. PMID- 12646479 TI - Bell-clapper deformity. PMID- 12646480 TI - Gallium uptake in complicated pancreatitis. PMID- 12646481 TI - CT screening examinations. PMID- 12646482 TI - Pitfall in interpretation of MR imaging of the bladder. PMID- 12646484 TI - Xanthoma disseminatum: a rare cause of upper airway narrowing. PMID- 12646485 TI - MR imaging of umbilical cord urachal (allantoic) cyst in utero. PMID- 12646486 TI - Chronic expanding hematoma of the thorax. PMID- 12646487 TI - Urachal mucinous cystadenocarcinoma with a cystic ovarian metastasis. PMID- 12646488 TI - Ubiquitination of prohibitin in mammalian sperm mitochondria: possible roles in the regulation of mitochondrial inheritance and sperm quality control. AB - Ubiquitination of the sperm mitochondria during spermatogenesis has been implicated in the targeted degradation of paternal mitochondria after fertilization, a mechanism proposed to promote the predominantly maternal inheritance of mitochondrial DNA in humans and animals. The identity of ubiquitinated substrates in the sperm mitochondria is not known. In the present study, we show that prohibitin, a highly conserved, 30- to 32-kDa mitochondrial membrane protein, occurs in a number of unexpected isoforms, ranging from 64 to greater than 185 kDa in the mammalian sperm mitochondria, which are the ubiquitinated substrates. These bands bind antiubiquitin antibodies, displaying a pattern consistent with polyubiquitinated "ladders." Immunoprecipitation of sperm extracts with antiprohibitin antibodies followed by probing of the resultant immunocomplexes with antiubiquitin yields a banding pattern identical to that observed by antiprohibitin Western blot analysis. In fact, the presumably nonubiquitinated 30-kDa prohibitin band shows no antiubiquitin immunoreactivity. We demonstrate that ubiquitination of prohibitin occurs in testicular spermatids and spermatozoa. Ubiquitinated prohibitin molecules also accumulate in the defective fractions of ejaculated spermatozoa, which are thought to undergo surface ubiquitination during epididymal passage. In such sperm fractions, ubiquitin also coprecipitates with tubulin and microtubule-associated proteins, presumably contributed by the axonemes of defective, ubiquitinated spermatozoa. The results of the present study suggest that prohibitin is one of the ubiquitinated substrates that makes the sperm mitochondria recognizable by the egg's ubiquitin-proteasome dependent proteolytic machinery after fertilization and most likely facilitates the marking of defective spermatozoa in the epididymis for degradation. PMID- 12646489 TI - Induction of a senescent-like phenotype does not confer the ability of bovine immortal cells to support the development of nuclear transfer embryos. AB - Previously, we reported that cloned embryos derived from an immortalized bovine mammary epithelial cell line (MECL) failed to develop beyond 12- to 16-cell stage. To analyze whether induction of a senescent-like phenotype in MECL can improve their ability to support the development after transfer into enucleated oocytes, we treated MECL with DNA methylation inhibitor 5-aza-2-deoxycytidine (Aza-C), histone deacetylase inhibitors trichostatin A (TSA), sodium butyrate (NaBu), or 5-bromodeoxyuridine and used those cells for nuclear transfer. Primary bovine fetal fibroblasts (BFF) were used as control. All agents were capable to induce features of senescence including reduced cell proliferation, enlarged cell size with a considerable proportion of cells stained positive for acidic senescence-associated beta-galactosidase and G1/S cell cycle boundary arrest in MECL. Aza-C treatment induced genome demethylation. Acetylation of H3 and H4 was increased after TSA treatment in both MECL and BFF, whereas no obvious changes in global H3 or H4 acetylation were detected after NaBu treatment. Nuclear transfer experiments following diverse treatments demonstrated that the induced senescent like phenotype of MECL did not confer their ability to support embryonic development, although 7.3% of reconstructed embryos derived from NaBu-treated cells developed to morula stage. Intriguingly, a much higher proportion of cloned embryos developed to blastocysts when using NaBu-treated BFF, compared with using untreated BFF (59% versus 26%). Our results suggest that the developmental failure of donor nuclei from bovine immortal cells could not be reversed by induction of senescent-like phenotype. The beneficial effect of NaBu on the developmental potential of cloned embryos reconstructed from BFF merits further studies. PMID- 12646490 TI - Relationship between endogenous progesterone and follicular dynamics in lactating dairy cows with ovarian follicular cysts. AB - Two experiments were conducted to examine circulating concentrations of progesterone (P4) in cows with ovarian follicular cysts (OFCs) and to relate differing levels of P4 to subsequent follicular events. In experiment 1, peripheral concentrations of P4 were determined in cows diagnosed with OFCs. Nonpregnant, lactating Holstein and Jersey cows (n = 32) were diagnosed as having OFCs by rectal palpation. Ovarian follicular cysts were then examined by transrectal ultrasonography to confirm the presence of OFCs (follicle diameter, >/=17 mm; absence of luteal tissue). At confirmation, a blood sample was collected for quantification of P4. The concentration of P4 at confirmation was classified as low (<0.1 ng/ml), intermediate (0.1-1.0 ng/ml), or high (1.0-2.0 ng/ml). More OFCs were associated with intermediate (66%) than with either low (28%) or high (6%) concentrations of P4. In experiment 2, the fate of follicles (diameter, >/=10 mm) that formed in the presence of an OFC was determined and related to circulating concentrations of P4 during follicular development. Follicles (n = 59) that formed in the presence of an OFC ovulated (n = 19), formed a cyst (n = 30), or underwent normal growth and regression (NGR; n = 10). Endogenous P4 in the 7-day period during follicular development was classified as low (if P4 dropped to <0.1 ng/ml for 1 day or longer), intermediate (if P4 averaged between 0.1 and 1.0 ng/ml and never dropped to <0.1 ng/ml), or high (if P4 averaged >1.0 ng/ml and never dropped to <0.1 ng/ml). In the presence of intermediate P4, 75% of observed follicles formed cysts, compared with 10% that ovulated and 15% that experienced NGR. In the presence of low P4, 53%, 41%, and 6% of follicles ovulated, formed a follicular cyst, or experienced NGR, respectively. Thus, an association between intermediate P4 and the formation of OFCs was established. PMID- 12646491 TI - Differential expression of the pituitary gonadotropin subunit genes during male rat sexual maturation: reciprocal relationship between hypothalamic pituitary adenylate cyclase-activating polypeptide and follicle-stimulating hormone beta expression. AB - The neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) has been shown to differentially regulate the expression of the gonadotropin subunit genes in cultures of rat pituitary cells. PACAP is expressed within the hypothalamus, and concentrations of PACAP are 2- to 4-fold higher in the portal circulation than in the general circulation. Therefore, PACAP is a candidate regulator of pituitary function. In the present study, we examined the expression of PACAP mRNA within the paraventricular nucleus (PVN) during maturation (Days 20 60) in the male rat and compared this expression to the levels of the gonadotropin subunits, follistatin, and GnRH-receptor mRNAs within the anterior pituitary. Serum concentrations of FSH and LH confirm the established maturational pattern of divergent secretion of LH and FSH. Northern analysis of the gonadotropin subunit mRNAs revealed that FSHbeta expression parallels FSH secretion whereas LHbeta mRNA concentrations do not change during development. Expression of the GnRH receptor in the pituitary parallels that of FSHbeta. In situ hybridization revealed a developmental pattern of PACAP mRNA within the PVN that is reciprocal to that of FSHbeta. Competitive reverse transcriptase polymerase chain reaction (RT-PCR) analysis of total pituitary follistatin mRNA revealed no significant changes; however, semiquantitative RT-PCR analyses revealed the presence of two follistatin mRNA species, one of which, corresponding to follistatin-288, was developmentally regulated. These studies identified a reciprocal relationship between PVN PACAP and FSHbeta gene expression in maturing rats. We propose that PACAP contributes to the selective regulation of FSHbeta expression during maturation in the male rat, perhaps via regulation of follistatin. PMID- 12646492 TI - Cytoskeleton and chromatin reorganization in horse oocytes following intracytoplasmic sperm injection: patterns associated with normal and defective fertilization. AB - Intracytoplasmic sperm injection (ICSI) is the method of choice for fertilizing horse oocytes in vitro. Nevertheless, for reasons that are not yet clear, embryo development rates are low. The aims of this study were to examine cytoskeletal and chromatin reorganization in horse oocytes fertilized by ICSI or activated parthenogenetically. Additional oocytes were injected with a sperm labeled with a mitochondrion-specific vital dye to help identify the contribution of the sperm to zygotic structures, in particular the centrosome. Oocytes were fixed at set intervals after sperm injection and examined by confocal laser scanning microscopy. In unfertilized oocytes, microtubules were present only in the metaphase-arrested second meiotic spindle and the first polar body. After sperm injection, an aster of microtubules formed adjacent to the sperm head and subsequently enlarged such that at the time of pronucleus migration and apposition it filled the entire cytoplasm. During syngamy, the microtubule matrix reorganized to form a mitotic spindle on which the chromatin of both parents aligned. Finally, after nuclear and cellular cleavage were complete, the microtubule asters dispersed into the interphase daughter cells. Sham injection induced parthenogenetic activation of 76% of oocytes, marked by the formation of multiple cytoplasmic microtubular foci that later developed into a dense microtubule network surrounding the female pronucleus. The finding that a parthenote alone can produce a microtubule aster, whereas the aster invariably forms at the base of the sperm head during normal fertilization, indicates that both gametes contribute to the formation of the zygotic centrosome in the horse. Finally, 25% of sperm-injected oocytes failed to complete fertilization, mostly due to absence of oocyte activation (65%), which was often accompanied by failure of sperm decondensation. In conclusion, this study demonstrated that union of the parental genomes in horse zygotes is accompanied by a series of integrated cytoskeleton-mediated events, failure of which results in developmental arrest. PMID- 12646493 TI - Deduction of a model for sperm storage in the oviduct of the domestic fowl (Gallus domesticus). AB - The mechanism of sperm storage in the fowl oviduct has remained a mystery since the 1960s, when sperm storage tubules (SST) were discovered between the shell gland and vagina. Previously, it was known that only motile sperm could ascend the vagina and enter these tubules. However, the means by which sperm resided therein was not clear. Research with computer-assisted sperm motion analysis has demonstrated that 1) seminal plasma glutamate acts as a motility agonist via N methyl-d-aspartate receptors; 2) motility depends on extracellular Ca2+ and Na+; 3) straight-line velocity is a variable with a skewed distribution; 4) sperm cell trajectory is a function of straight-line velocity; and 5) specific inhibition of phospholipase A2 renders sperm immotile. An additional experiment demonstrated that Ca2+ acts as a second messenger and thereby modulates the content of long chain acylcarnitine within sperm. Therefore, it is proposed that 1) the release of endogenous fatty acids fuels sperm as they ascend the vagina; (2) on entering the SST, motile sperm maintain position against a fluid current generated by SST epithelial cells; 3) resident sperm metabolize exogenous fatty acids released from lipid-laden epithelial cells; (4) motile sperm emerge from the SST when their velocity declines to a threshold at which retrograde movement begins; and 5) the skewed distribution of straight-line velocity accounts for the exponential pattern of sperm emergence from the SST. In summary, sperm residence within and emergence from the SST are phenomena most likely explicable in terms of sperm cell motility. PMID- 12646494 TI - Complementary DNA cloning and characterization of rat spergen-2, a spermatogenic cell-specific gene 2 encoding a 56-kilodalton nuclear protein bearing ankyrin repeat motifs. AB - Differential display in combination with a cDNA cloning approach were used to isolate a novel gene, spergen-2, which has an open reading frame of 1500 nucleotides and encodes a protein of 500 amino acids that contains ankyrin repeat motifs and a putative nuclear localization signal. Expression of spergen-2 is developmentally upregulated in testis. In situ hybridization revealed that spergen-2 mRNA is expressed in spermatocytes and round spermatids (steps 1-6). Immunohistochemical analysis with confocal laser-scanning microscopy demonstrated that spergen-2 protein is predominantly expressed in nuclei of late spermatocytes (stages IX-XIV) and spermatids (steps 1-11), indicating the restricted expression of spergen-2 during spermatogenesis. In nucleoplasm of spermatogenic cell nuclei, spergen-2 tends to localize in the interchromosome space with relatively low DNA density. These findings indicate a potential role of spergen-2 in spermatogenesis, especially in cell differentiation from late pachytene spermatocytes to spermatids or in early spermatid differentiation. PMID- 12646495 TI - Analysis of cytokine regulators inducing interferon production by mouse uterine natural killer cells. AB - In mice and women, terminal differentiation of uterine natural killer (uNK) cells commences during endometrial decidualization. Both proliferation and interferon (IFN)-gamma are induced. Uterine NK cell precursors appear to home from secondary lymphoid organs to decidualizing uteri and localize mesometrially to the central decidua basalis, the site of maternal arterial modification at Gestation Days (gd) 9.5-10. In mice, genetic absence of uNK cells results in absence of pregnancy-induced spiral artery modification. Administration of IFN-gamma to uNK negative pregnant females induces arterial modifications without fetal loss. In this study, we investigated the roles of cytokines, known in other tissues to differentiate and activate NK cells, in induction of IFN-gamma production in normal mouse implantation sites. Fecundity evaluation, implantation site morphometry, and IFN-gamma quantification in interleukin (IL)-12p40(0/0), IL 18(0/0), dual IL-12p40(0/0)/IL-18(0/0) and congenic strains revealed the importance of both IL-12 and IL-18 in the induction of spiral artery modification and IFN-gamma synthesis. Immediately after implantation, IL-18 was localized transiently to decidual cells, but by gd8, IL-18 was produced solely by uNK cells, suggesting that early uNK cells are activated by stroma and lymphocyte derived signals maintain later uNK cell activation. Mesometrial tissue of C57Bl/6J mice was examined by reverse transcription polymerase chain reaction assay in virgin, early postimplantation, and midgestation females for expression of the heterodimeric cytokines IL-23 (composed of IL-12p40 and a novel alpha chain), IL-27 (composed of two IL-12-related chains) and IL-27R. No expression was detected in virgin uteri. The four genes were induced by gd6, and uNK cells isolated from midgestation transcribed IL-23alpha and IL-27R. This study advances the understanding of uNK cell activation during normal pregnancy. PMID- 12646497 TI - Does canopy position affect wood specific gravity in temperate forest trees? AB - The radial increases in wood specific gravity known in many tree species have been interpreted as providing mechanical support in response to the stresses associated with wind loading. This interpretation leads to the hypothesis that individuals reaching the canopy should (1) be more likely to have radial increases in specific gravity and (2) exhibit greater increases than individuals in the subcanopy. Wood specific gravity was determined for three species of forest trees (Acer rubrum, Fagus grandifolia and Tsuga canadensis) growing in central Massachusetts, USA. Acer rubrum shows radial increases in specific gravity, but these increases are not more pronounced in canopy trees; the other two species show a pattern of radial decreases. The degree of radial increase or decrease is influenced by tree height and diameter. Of the dominant tree species for which we have data, A. rubrum, Betula papyrifera and Pinus strobus show radial increases in specific gravity, whereas F. grandifolia, T. canadensis and Quercus rubra show decreases. The occurrence of radial increases in B. papyrifera and P. strobus, which are often canopy emergents, suggests that it is overall adaptive strategy that is important rather than position (canopy vs. subcanopy) of any individual tree. It is suggested that radial increases in specific gravity are associated with early-successional status or characteristics and decreases with late-successional status or persistence in mature forest. PMID- 12646496 TI - Na+ tolerance and Na+ transport in higher plants. AB - Tolerance to high soil [Na(+)] involves processes in many different parts of the plant, and is manifested in a wide range of specializations at disparate levels of organization, such as gross morphology, membrane transport, biochemistry and gene transcription. Multiple adaptations to high [Na(+)] operate concurrently within a particular plant, and mechanisms of tolerance show large taxonomic variation. These mechanisms can occur in all cells within the plant, or can occur in specific cell types, reflecting adaptations at two major levels of organization: those that confer tolerance to individual cells, and those that contribute to tolerance not of cells per se, but of the whole plant. Salt tolerant cells can contribute to salt tolerance of plants; but we suggest that equally important in a wide range of conditions are processes involving the management of Na(+) movements within the plant. These require specific cell types in specific locations within the plant catalysing transport in a coordinated manner. For further understanding of whole plant tolerance, we require more knowledge of cell-specific transport processes and the consequences of manipulation of transporters and signalling elements in specific cell types. PMID- 12646498 TI - Seasonal patterns of 13C partitioning between shoots and nodulated roots of N2- or nitrate-fed Pisum sativum L. AB - The effect of nitrogen source (N(2) or nitrate) on carbon assimilation by photosynthesis and on carbon partitioning between shoots and roots was investigated in pea (Pisum sativum L. 'Baccara') plants at different growth stages using (13)C labelling. Plants were grown in the greenhouse on different occasions in 1999 and 2000. Atmospheric [CO(2)] and growth conditions were varied to alter the rate of photosynthesis. Carbon allocation to nodulated roots was unaffected by N source. At the beginning of the vegetative period, nodulated roots had priority for assimilates over shoots; this priority decreased during later stages and became identical to that of the shoot during seed filling. Carbon allocation to nodulated roots was always limited by competition with shoots, and could be predicted for each phenological stage: during vegetative and flowering stages a single, negative exponential relationship was established between sink intensity (percentage of C allocated to the nodulated root per unit biomass) and net photosynthesis. At seed filling, the amount of carbon allocated to the nodulated root was directly related to net photosynthesis. Respiration of nodulated roots accounted for more than 60 % of carbon allocated to them during growth. Only at flowering was respiration affected by N supply: it was significantly higher for strictly N(2)-fixing plants (83 %) than for plants fed with nitrate (71 %). At the vegetative stage, the increase in carbon in nodulated root biomass was probably limited by respiration losses. PMID- 12646499 TI - Comparisons with Caenorhabditis (approximately 100 Mb) and Drosophila (approximately 175 Mb) using flow cytometry show genome size in Arabidopsis to be approximately 157 Mb and thus approximately 25% larger than the Arabidopsis genome initiative estimate of approximately 125 Mb. AB - Recent genome sequencing papers have given genome sizes of 180 Mb for Drosophila melanogaster Iso-1 and 125 Mb for Arabidopsis thaliana Columbia. The former agrees with early cytochemical estimates, but numerous cytometric estimates of around 170 Mb imply that a genome size of 125 Mb for arabidopsis is an underestimate. In this study, nuclei of species pairs were compared directly using flow cytometry. Co-run Columbia and Iso-1 female gave a 2C peak for arabidopsis only approx. 15 % below that for drosophila, and 16C endopolyploid Columbia nuclei had approx. 15 % more DNA than 2C chicken nuclei (with >2280 Mb). Caenorhabditis elegans Bristol N2 (genome size approx. 100 Mb) co-run with Columbia or Iso-1 gave a 2C peak for drosophila approx. 75 % above that for 2C C. elegans, and a 2C peak for arabidopsis approx. 57 % above that for C. elegans. This confirms that 1C in drosophila is approx. 175 Mb and, combined with other evidence, leads us to conclude that the genome size of arabidopsis is not approx. 125 Mb, but probably approx. 157 Mb. It is likely that the discrepancy represents extra repeated sequences in unsequenced gaps in heterochromatic regions. Complete sequencing of the arabidopsis genome until no gaps remain at telomeres, nucleolar organizing regions or centromeres is still needed to provide the first precise angiosperm C-value as a benchmark calibration standard for plant genomes, and to ensure that no genes have been missed in arabidopsis, especially in centromeric regions, which are clearly larger than once imagined. PMID- 12646500 TI - Influence of elevated CO2 and O3 on Betula pendula Roth crown structure. AB - Elevated CO(2) and ozone effects were studied singly and in combination on the crown structure of two Betula pendula clones. Measurements were made at the end of the second fumigation period in an open-top-chamber experiment with 9-year-old trees. Shoot ramification (number of long and short daughter shoots), shoot length, and number of metamers, leaves and buds were measured at four positions in every tree. As a result of increased temperature, trees in chambers had longer shoots and more frequent shoot ramification than control trees not enclosed in chambers. Ozone treatment decreased shoot ramification significantly. Additionally, ozone treatment resulted in an increased number of metamers in one clone. There was no statistically significant interaction between ozone effect and crown position; however, there was a slight tendency for the lower crown to be more affected by ozone. Elevated CO(2) caused a significant increase in the number of long-shoot metamers. Therefore, 2x ambient CO(2) concentration partly ameliorated the negative effect of ozone because the increased number of leaves per shoot counteracted the decreased branching. Although the main effects of elevated ozone and CO(2) were similar in the two clones, slight, statistically insignificant, differences appeared in their responses when interactions with crown position were considered. PMID- 12646501 TI - Cell division and morphological changes in the shoot apex of Arabidopsis thaliana during floral transition. AB - Eight-week-old vegetative plants of Arabidopsis thaliana, ecotype Columbia, were induced to flower by a single long day (LD). In this experimental system, it is known that the last component of the floral stimulus moves from the leaves to the apex 24-36 h after the start of the LD, and the first floral meristem is initiated by the shoot apical meristem (SAM) at 44-56 h (Corbesier et al., 1996, The Plant Journal 9: 947-952). Here we show that the rate of cell division is increased at floral transition in all SAM parts but not in the sub-apical pith cells. Mitotic activity starts to increase 24 h after the start of the LD and is two- to three-fold higher at peak times than that in non-induced plants. This activation is followed by the start of SAM enlargement at 44 h, SAM doming at 48 h, and the elongation of apical internodes (bolting) at 52 h. PMID- 12646502 TI - The ratio of leaf to total photosynthetic area influences shade survival and plastic response to light of green-stemmed leguminous shrub seedlings. AB - Different plant species and organs within a plant differ in their plastic response to light. These responses influence their performance and survival in relation to the light environment, which may range from full sunlight to deep shade. Plasticity, especially with regard to physiological features, is linked to a greater capacity to exploit high light and is usually low in shade-tolerant species. Among photosynthetic organs, green stems, which represent a large fraction of the total photosynthetic area of certain species, are hypothesized to be less capable of adjustment to light than leaves, because of biomechanical and hydraulic constraints. The response to light by leaves and stems of six species of leguminous, green-stemmed shrubs from dry and high-light environments was studied by growing seedlings in three light environments: deep shade, moderate shade and sun (3, 30 and 100 % of full sunlight, respectively). Survival in deep shade ranged from 2 % in Retama sphaerocarpa to 74 % in Ulex europaeus. Survival was maximal at moderate shade in all species, ranging from 80 to 98 %. The six species differed significantly in their ratio of leaf to total photosynthetic area, which influenced their light response. Survival in deep shade increased significantly with increasing ratio of leaf to total photosynthetic area, and decreased with increasing plasticity in net photosynthesis and dark respiration. Responses to light differed between stems and leaves within each species. Mean phenotypic plasticity for the variables leaf or stem specific mass, chlorophyll content, chlorophyll a/b ratio, and carotenoid to chlorophyll ratio of leaves, was inversely related to that of stems. Although mean plasticity of stems increased with the ratio of leaf to total photosynthetic area, the mean plasticity of leaves decreased. Shrubs with green stems and a low ratio of leaf to total photosynthetic area are expected to be restricted to well-lit habitats, at least during the seedling stage, owing to their inefficient light capture and the low plasticity of their stems. PMID- 12646503 TI - Dynamics of endogenous cytokinin pools in tobacco seedlings: a modelling approach. AB - Recent advances in cytokinin analysis have made it possible to measure the content of 22 cytokinin metabolites in the tissue of developing tobacco seedlings. Individual types of cytokinins in plants are interconverted to their respective forms by several enzymatic activities (5'-AMP-isopentenyltransferase, adenosine nucleosidase, 5'-nucleotidase, adenosine phosphorylase, adenosine kinase, trans-hydroxylase, zeatin reductase, beta-glucosidase, O-glucosyl transferase, N-glucosyl transferase, cytokinin oxidase). This paper reports modelling and measuring of the dynamics of endogenous cytokinins in tobacco plants grown on media supplemented with isopentenyl adenine (IP), zeatin (Z) and dihydrozeatin riboside (DHZR). Differences in phenotypes generated by the three cytokinins are shown and discussed, and the assumption that substrate concentration drives enzyme kinetics underpinned the construction of a simple mathematical model of cytokinin metabolism in developing seedlings. The model was tested on data obtained from liquid chromatography/tandem mass spectrometry cytokinin measurements on tobacco seedlings grown on Murashige and Skoog agar nutrient medium, and on plants grown in the presence of IP, Z and DHZR. A close match was found between measured and simulated data, especially after a series of iterative parameter searches, in which the parameters were set to obtain the best fit with one of the data sets. PMID- 12646504 TI - Over-expression of Arabidopsis CAP causes decreased cell expansion leading to organ size reduction in transgenic tobacco plants. AB - Cyclase-associated proteins (CAP) are multifunctional proteins involved in Ras cAMP signalling and regulation of the actin cytoskeleton. It has recently been demonstrated that over-expression of AtCAP1 in transgenic arabidopsis plants causes severe morphological defects owing to loss of actin filaments. To test the generality of the function of AtCAP1 in plants, transgenic tobacco plants over expressing an arabidopsis CAP (AtCAP1) under the regulation of a glucocorticoid inducible promoter were produced. Over-expression of AtCAP1 in transgenic tobacco plants led to growth abnormalities, in particular a reduction in the size of leaves. Morphological alterations in leaves were the result of reduced elongation of epidermal and mesophyll cells. PMID- 12646506 TI - Personal exposure to PM2.5 and biomarkers of DNA damage. AB - Ambient particulate air pollution assessed as outdoor concentrations of particulate matter < or = 2.5 microm in diameter (PM(2.5)) has been associated with an increased cancer risk. However, outdoor PM(2.5) concentrations may not be the best measure of the individual particle exposure that is a sum of many sources besides outdoor particle levels, e.g., environmental tobacco smoke and cooking. We measured personal PM(2.5) and black smoke exposure in 50 students four times over 1 year and analyzed for biomarkers of different types of DNA damages. Ambient PM(2.5) concentrations were also measured. Exposure was measured for 48 h, after which blood samples were collected and analyzed for DNA damage in lymphocytes in terms of 7-hydro-8-oxo-2'-deoxyguanosine (8-oxodG), strand breaks, endonuclease III- and fapyguanine glycosylase-sensitive sites, and polyaromatic hydrocarbon adducts. Twenty-four-h urine collections were analyzed for 8-oxodG and 1-hydroxypyrene. Personal PM(2.5) exposure was found to be a predictor of 8 oxodG in lymphocyte DNA with an 11% increase in 8-oxodG/10 microg/m(3) increase in personal PM(2.5) exposure (P = 0.007). No other associations between exposure markers and biomarkers could be distinguished. The genotype of glutathione S transferase M1 (GSTM1), T1 (GSTT1), and P1 (GSTP1) and NADPH:quinone reductase was also determined, but there were no effects of genotype on DNA polyaromatic hydrocarbon adducts or oxidative damage. The results suggest that moderate exposure to concentrations of PM can induce oxidative DNA damage and that personal PM(2.5) exposure is more important in this aspect than is ambient PM(2.5) background concentration. PMID- 12646505 TI - Interaction between CYP1A1 polymorphic variants and dietary exposures influencing ovarian cancer risk. AB - Aromatic hydrocarbon hydroxylase (CYP1A1) is involved in the metabolism of many substrates and the subject of cancer studies. This study examined the association between two polymorphic variants of CYP1A1 and ovarian cancer risk. The frequencies of the Msp1 and Ile/Val variants of CYP1A1 were determined in 445 ovarian cancer cases and 472 general population controls in New England. Overall relative risks were calculated as well as those within subgroups of various exposures. There was no increased risk for ovarian cancer associated with possession of either the Msp1 or Ile/Val polymorphism of CYP1A1. Elevated risk for ovarian cancer was found in those who carried an Ile/Val variant and who consumed more than median levels of caffeine (risk ratio = 2.69; 95% confidence interval, 1.18-6.18). No variation by histological type of ovarian cancer was observed. Significant interaction may exist between polymorphic variants of CYP1A1 and caffeine that could explain weak or inconsistent associations between caffeine and ovarian cancer when genotype has not been considered. PMID- 12646507 TI - Fecal bile acid concentrations in a subpopulation of the wheat bran fiber colon polyp trial. AB - Factors that affect the concentration of secondary bile acids in the aqueous phase of stool may have a greater impact on colon carcinogenesis than those that only modify the total fecal bile acid concentration. This hypothesis was tested using stool samples of a subset of participants enrolled in a Phase III colorectal adenomatous polyp prevention trial, which documented the inability of a 13.5 g/day wheat bran fiber (WBF) supplement to reduce polyp recurrence. Stool was collected from 68 consecutively consented participants who were enrolled in a Phase III clinical trial of WBF for the prevention of adenomatous polyp recurrence. Nineteen (27.9%) of these fecal bile acid substudy participants were on the low fiber (2.0 g/day) intervention group, whereas 49 (72.7%) were on the high fiber (13.5 g/day) intervention group for approximately 3 years. Sixty-four participants had both the aqueous and solid phases of stool samples analyzed for bile acid content. Bile acid concentrations, measured in microg/ml for fecal water and microg/mg for dry feces, were determined for lithochilic, deoxycholic, chenodeoxycholic, cholic, ursodeoxycholic, isodeoxycholic, isoursodeoxycholic, ursocholic, 7-ketolithocholic, and 12-ketolithocholic acids. There were no significant differences between the low and high fiber groups concerning mean or median aqueous phase concentrations of lithocholic or deoxycholic bile acids. In contrast, the median concentrations of deoxycholic acid and other secondary bile acids (including lithochilic, isodeoxycholic, ursodeoxycholic, isoursodeoxycholic, ursocholic, 7-ketolithocholic, and 12-ketolithocholic acids) were significantly lower for the high fiber group in the solid-phase stool (P < 0.05). These results document that a high WBF intervention, taken for a median of 2.4 years, does not significantly reduce aqueous-phase concentrations of secondary bile acids in stool, although their concentrations in solid-phase stool were suppressed. Thus, the inability of the high WBF intervention to reduce colorectal adenoma recurrence may be a consequence of its lack of effect on fecal aqueous-phase secondary bile acid concentrations. PMID- 12646508 TI - Dietary factors and risk of colon cancer in Shanghai, China. AB - Colon cancer incidence rates have risen sharply in Shanghai, China, since the early 1970s, and diet may have contributed to the rising incidence. To clarify the role of dietary factors for colon cancer in Shanghai, we analyzed data from a population-based case-control study of 931 cases (462 males and 469 females) and 1552 controls (851 males and 701 females) ages 30-74 years in Shanghai, China, from 1990-1993. Subjects were interviewed in person for a detailed history of dietary practices and food preferences by using a food-frequency questionnaire. Colon cancer risk was estimated by odds ratios (ORs) and 95% confidence intervals (CIs), adjusting for age, total energy, and other confounding factors. Risk for the highest versus the lowest quartile of intake was elevated for red meat (OR, 1.5; 95% CI, 1.0-2.1 for men and OR, 1.5; 95% CI, 1.0-2.2 for women), fish (OR, 1.7; 95% CI, 1.2-2.4 for men and OR, 1.2; 95% CI, 0.8-1.7 for women), and eggs (OR, 1.4; 95% CI, 1.0-1.9 for men and OR, 1.3; 95% CI, 0.9-1.9 for women), but was reduced for fresh fruit (OR, 0.7; 95% CI, 0.5-1.0 for men and OR, 0.6, 0.4 0.9 for women). High intake of preserved foods, whether animal or plant source, was associated with an excess risk of colon cancer (OR, 2.0; 95% CI, 1.5-2.9 for men and OR, 2.7; 95% CI, 1.9-3.8 for women). For dietary nutrients, risk generally declined with greater consumption of fiber and micronutrients common in fruit and vegetables, including vitamin C, carotene, and vitamin E. Intake of macronutrients in general was not significantly related to risk. Our findings suggest that diets high in fruit and antioxidant vitamins that are common in plant foods reduce the risk of colon cancer, whereas diets high in red meat, eggs, and preserved foods increase the risk. PMID- 12646509 TI - Induced abortion, miscarriage, and breast cancer risk of young women. AB - Early studies of breast cancer raised substantial concern regarding risk associated with induced abortion and miscarriage. Literature reviews suggest that study findings depend heavily on the comparison group and that the use of parous women as a reference group for nulliparous women may artificially inflate risk. To examine the individual effects of induced abortion and miscarriage on breast cancer risk of parous and nulliparous women, 744 patients < or =40 years of age and diagnosed from 1983-1988 were matched by parity, age, and race with controls living in the same neighborhood in Los Angeles County. In-person interviews were conducted to obtain a detailed reproductive history. Risk estimates were obtained by conditional logistic regression using nulligravid women as the reference group for nulliparous women with a history of incomplete pregnancy and parous women with no incomplete pregnancies as the reference group for parous women with a history of incomplete pregnancy. Breast cancer risk of parous women was unrelated to a history of miscarriage or induced abortion. Breast cancer risk was reduced among nulliparous women with a history of induced abortion relative to nulligravid women, although the risk estimate was imprecise. Risk declined as the number of induced abortions increased (P = 0.04). Our results do not support the hypothesis that induced abortion or miscarriage increase the breast cancer risk of young women. PMID- 12646510 TI - Height and the survival of prostate cancer patients. AB - We investigated the associations between height and other anthropometric factors and the survival of 584 prostate cancer patients, initially recruited for a population-based, case-control study. During a median of 6.6 years of follow-up, 129 prostate cancer deaths and 153 deaths because of other causes were identified. After adjusting for age, cancer stage, and grade, the relative risk and 95% confident intervals for prostate cancer death were 1.0 (reference), 0.9 (0.6-1.4), 0.5 (0.3-0.9), and 0.6 (0.3-1.0) for patients whose heights were <1.75 m, 1.75-1.79 m, 1.80-1.84 m, and > or =1.85 m, respectively (P for trend = 0.01). Similar associations were found in subgroup analyses by cancer stage, cancer grade, age, race, and occupation-based socioeconomic status. However, height was not associated with death because of other causes. In addition, no significant associations were found between body mass index or weight and either prostate cancer death or death because of other causes. Our results suggest that greater height may be associated with better survival of prostate cancer patients. PMID- 12646511 TI - Aryl hydrocarbon hydroxylase represents CYP1B1, and not CYP1A1, in human freshly isolated white cells: trimodal distribution of Japanese population according to induction of CYP1B1 mRNA by environmental dioxins. AB - The expression level of mRNAs for cytochrome P450 (CYP) 1A1 and 1B1 in freshly prepared white cells from 72 subjects exposed to dioxins at waste incinerators was investigated. The amounts of CYP1B1 mRNA ranged from 0.16 to 671 molecules/10(7) molecules of 18S rRNA, whereas the amounts of CYP1A1 mRNA were <6 molecules/10 ng total RNA, indicating that CYP1A1 was not induced to a detectable level by environmentally exposed dioxins. The inducibility of CYP1B1 mRNA in leukocytes, defined as the ratio of CYP1B1 mRNA to the plasma concentration of dioxins, varied among the subjects. It was found that the subjects showed trimodal distribution according to inducibility: 39 (54.2%), 25 (34.7%), and 8 (11.1%) of 72 subjects were judged as poor, intermediate, and high responders to environmental dioxins, respectively. The amounts of CYP1B1 mRNA in leukocytes of the intermediate and high responders were highly correlated with the plasma concentrations of dioxins (P < 0.05 and <0.01). These results suggest that CYP1B1 with polymorphic inducibility by dioxins is involved in aromatic hydrocarbon hydroxylase activities in human lymphocytes. PMID- 12646512 TI - Handedness and risk of brain tumors in adults. AB - The objective of this study was to evaluate the relation between handedness, and the risk of malignant and benign brain tumors. Handedness has been hypothesized to serve as a behavioral marker of prenatal hormonal exposures or other factors that influence subsequent cancer risk. A case-control study was conducted at hospitals in three United States cities between 1994 and 1998. The cases were adult patients newly diagnosed with glioma (n = 489), meningioma (n = 197), or acoustic neuroma (n = 96), and the 799 frequency-matched controls were patients admitted to the same hospitals for a variety of nonmalignant conditions. Handedness was determined by interview. Unconditional logistic regression was used to estimate odds ratios (ORs) and calculate 95% confidence intervals (CIs). Persons who described themselves as left-handed or ambidextrous appeared to be at reduced risk of glioma relative to those who described themselves as right-handed (OR, 0.7; 95% CI, 0.5-0.9). The association was similar for men and women, and for left-sided and right-sided tumors. Neither meningioma (OR, 0.9; CI, 0.6-1.5) nor acoustic neuroma (OR, 0.9; CI, 0.5-1.7) showed significant associations with handedness. These findings require confirmation but raise the possibility that early neurodevelopmental events or genetic factors related to handedness also influence the risk of glioma among adults. PMID- 12646513 TI - No relationship between ovarian cancer risk and progesterone receptor gene polymorphism in a population-based, case-control study in North Carolina. PMID- 12646514 TI - Proceedings of a symposium on reducing the unequal burden of cancer. Boston, Massachusetts, USA. PMID- 12646515 TI - Reducing the unequal burden of cancer. PMID- 12646516 TI - Commentary on the meaning of race in science and society. PMID- 12646517 TI - Cancers caused by infections: unequal burdens. PMID- 12646518 TI - Helicobacter pylori infection and gastric cancer. PMID- 12646519 TI - Cervical cancer: disparities in screening, treatment, and survival. PMID- 12646520 TI - Viral hepatitis and hepatocellular carcinoma in African Americans. PMID- 12646521 TI - Cancer prevention and early detection. PMID- 12646522 TI - Challenges in tobacco use prevention among minority youth. PMID- 12646523 TI - Reducing health disparities: from theory to practice. PMID- 12646524 TI - The role of state health agencies in cancer prevention and control: lessons learned from Massachusetts. PMID- 12646525 TI - Cancer therapy and survivorship. PMID- 12646526 TI - Reducing the burden of cancer borne by African Americans: if not now, when? PMID- 12646527 TI - Increasing participation of physicians and patients from underrepresented racial and ethnic groups in National Cancer Institute-sponsored clinical trials. PMID- 12646528 TI - Cancer survivorship issues for minority and underserved populations. PMID- 12646531 TI - "No ball games": children's views of their urban environments. PMID- 12646533 TI - The caterpillar and the butterfly: public health in Spain. PMID- 12646534 TI - Public health in the new European Union research programme. PMID- 12646535 TI - Media coverage as a risk factor in suicide. AB - A total of 293 findings from 42 studies on the impact of publicized suicide stories in the media on the incidence of suicide in the real world were analyzed by logistic regression analysis. Studies measuring the effect of either an entertainment or political celebrity suicide story were 14.3 times more likely to find a copycat effect than studies that did not. Studies based on a real as opposed to fictional story were 4.03 times more likely to uncover a copycat effect. Research based on televised stories was 82% less likely to report a copycat effect than research based on newspapers. A review of recent events in Austria and Switzerland indicates that suicide prevention organizations can successfully convince the media to change the frequency and content of their suicide coverage in an effort to reduce copycat effects. PMID- 12646536 TI - Who decides when to start preventive treatment? A questionnaire survey to compare the views of different population subgroups. PMID- 12646537 TI - Substantial potential for reductions in coronary heart disease mortality in the UK through changes in risk factor levels. AB - STUDY OBJECTIVE: The UK government called for a 40% reduction in cardiovascular disease mortality in those aged under 75 by 2010. This paper examines the potential for cardiovascular risk factor changes to reduce coronary heart disease deaths in Scotland, and then extrapolates the findings to the UK population. DESIGN: Secondary analysis of published data using a previously validated mortality model. The model combines uptake and effectiveness of treatments with risk factor trends by sex and age group. It was used to estimate the expected reductions in coronary heart disease mortality: (a) if recent risk factor trends simply continued; (b) if additional risk factor reductions were achieved in line with Scandinavia and the United States. An "analysis of extremes" sensitivity analysis was then carried out. SETTING: Scotland and UK. PARTICIPANTS: Projected Scottish population aged 45+ in 2010 (2.4 million) and UK population of 26.8 million. MAIN RESULTS: Continuation of current trends would result in 2169 fewer coronary deaths in 2010 (minimum estimate 1191 from sensitivity analyses to maximum 3870). About 4749 fewer deaths (minimum 3085, maximum 7155) could be achieved by: (a) a reduction in smoking prevalence from 30% to 18% (about 1668 fewer deaths); (b) a mean population cholesterol reduction from 6.2 to 5.2 mmol/l (about 2167 fewer deaths); (c) a 3.7 mm Hg fall in diastolic blood pressure (about 914 fewer deaths). Extrapolation from the Scottish population to the UK suggests 24 000 fewer deaths in 2010 if current trends continue, or 53 000 fewer deaths with the additional reductions. CONCLUSIONS: With additional interventions it would be possible to almost halve current UK coronary heart disease mortality. Even without gains from medical treatments, the UK government target of 28 000 fewer deaths in 2010 does not seem challenging. PMID- 12646538 TI - Measuring the habitat as an indicator of socioeconomic position: methodology and its association with hypertension. AB - STUDY OBJECTIVES: (1) to develop an indicator of socioeconomic position based on the social standing of the habitat (SSH), that is, the residential building, its immediate surroundings, and local neighbourhood; (2) to assess the relation of SSH to two usual markers of socioeconomic position (education and occupation) and a known, socially determined health outcome (hypertension). DESIGN: Population survey measuring SSH, detailed educational and occupational histories, and blood pressure. The SSH is a standardised assessment of the external and internal aspects of someone's building (or house), and of the characteristics of its immediate surroundings and local neighbourhood. SETTING: A sample of participants to the Bus Sante survey between 1993 and 1998, in Geneva, Switzerland. PARTICIPANTS: 588 men and women, aged 35 to 74. MAIN RESULTS: The SSH index was highly reproducible (kappa=0.8). Concordance of SSH with education or occupation was good for people of either high or low socioeconomic position, but not for those with medium education and/or occupation. There was a higher prevalence of hypertension in the lowest compared with the highest groups, defined on the basis of education or occupation, but the SSH was the only indicator that showed a higher prevalence of hypertension among people in the middle of the social spectrum. CONCLUSIONS: People of medium education or occupation are heterogeneous with respect to their habitat. Those living in habitats of medium social standing may be most affected by hypertension but this association could not be revealed on the basis of education and occupation alone. The habitat seems to capture different aspects of the socioeconomic position compared with the usual indicators of social class. PMID- 12646540 TI - Chinese SF-36 Health Survey: translation, cultural adaptation, validation, and normalisation. AB - STUDY OBJECTIVE: To develop a self administered Chinese (mainland) version of the Short-Form Health Survey (SF-36) for use in health related quality of life measurements in China. DESIGN: A three stage protocol was followed including translation, tests of scaling construction and scoring assumptions, validation, and normalisation. SETTING: 1000 households in 18 communities of Hangzhou. PARTICIPANTS: 1688 respondents recruited by multi-stage mixed sampling. MAIN RESULTS: The assumption of equal intervals was violated for the vitality and mental health scales. The recoded item values were used to calculate scale scores. The clustering and ordering of item means was the same as that of the source and other two Chinese versions. The items in each scale had similar standard deviations except those in the physical functioning, boduily pain, social functioning scales. The item hypothesised scale correlations were identical for all except the social functioning and vitality scales. Convergent validity and discriminant validity were satisfactory for all except the social functioning scale. Cronbach's alpha coefficients ranged from 0.72 to 0.88 except 0.39 for the social functioning scale and 0.66 for the vitality scale. Two weeks test-retest reliability coefficients ranged from 0.66 to 0.94. Factor analysis identified two principal components explaining 56.3% of the total variance. The Chinese SF-36 could distinguish known groups. CONCLUSIONS: This study suggested that the Chinese (mainland) version of the SF-36 functioned in the general population of Hangzhou, China quite similarly to the original American population tested. Caution is recommended in the interpretation of the social functioning and vitality scales pending further studies. PMID- 12646539 TI - Defining equity in health. AB - STUDY OBJECTIVE: To propose a definition of health equity to guide operationalisation and measurement, and to discuss the practical importance of clarity in defining this concept. DESIGN: Conceptual discussion. Setting, Patients/Participants, and Main results: not applicable. CONCLUSIONS: For the purposes of measurement and operationalisation, equity in health is the absence of systematic disparities in health (or in the major social determinants of health) between groups with different levels of underlying social advantage/disadvantage-that is, wealth, power, or prestige. Inequities in health systematically put groups of people who are already socially disadvantaged (for example, by virtue of being poor, female, and/or members of a disenfranchised racial, ethnic, or religious group) at further disadvantage with respect to their health; health is essential to wellbeing and to overcoming other effects of social disadvantage. Equity is an ethical principle; it also is consonant with and closely related to human rights principles. The proposed definition of equity supports operationalisation of the right to the highest attainable standard of health as indicated by the health status of the most socially advantaged group. Assessing health equity requires comparing health and its social determinants between more and less advantaged social groups. These comparisons are essential to assess whether national and international policies are leading toward or away from greater social justice in health. PMID- 12646541 TI - Landmine related injuries in children of Bosnia and Herzegovina 1991-2000: comparisons with adults. PMID- 12646542 TI - Educational level as a contextual and proximate determinant of all cause mortality in Danish adults. AB - STUDY OBJECTIVE: To examine the educational level in the area of living as a determinant of all cause mortality, controlling for individual and other correlated contextual factors. DESIGN: Pooled data from two population based cohort studies were linked to social registers to obtain selected socioeconomic information at parish and individual level. A total of 18 344 men and women were followed up from 1980 until October 1999. SETTING: Copenhagen, Denmark. MAIN OUTCOME MEASURE: All cause mortality. RESULTS: During follow up 2614 men and women died. Educational status both at parish (hazard ratio (HR): 0.87 (95% CI 0.77 to 0.98) and individual level (HR: 0.76 (95% CI 0.64 to 0.88) were inversely associated with mortality, when comparing the higest educated groups with the least educated. However, at parish level the effect was only present, when information on subject's income, behaviour (smoking, exercise, alcohol use, and body mass index) and contextual factors (local area unemployment, income share, and household composition) were included in the Cox model. CONCLUSION: In this study the educational level of an area influenced subject's mortality, but first after adjustment for behavioural and other contextual risk factors. Neighbourhood education is one of different characteristics of adverse social conditions in an area increasing mortality. PMID- 12646544 TI - The historical specificity of early life car ownership as an indicator of socioeconomic position. PMID- 12646543 TI - Residential area deprivation predicts smoking habit independently of individual educational level and occupational social class. A cross sectional study in the Norfolk cohort of the European Investigation into Cancer (EPIC-Norfolk). AB - OBJECTIVES: To investigate the independent association between individual and area based measures of socioeconomic status and cigarette smoking habit. DESIGN AND SETTING: Cross sectional, population based study. PARTICIPANTS AND METHODS: 12 579 men and 15 132 women aged 39-79 years living in the general community participating in the EPIC-Norfolk Study in 1993-1997. The association between social class, educational status, Townsend residential deprivation level, and cigarette smoking status was examined. MAIN OUTCOME MEASURES: Cigarette smoking status at baseline survey. RESULTS: Social class, educational level, and residential deprivation level independently related to cigarette smoking habit in both men and women. Multivariate age adjusted odds ratios for current smoking in men were 1.62 (95% CI 1.45 to 1.81) for manual compared with non-manual social class, 1.32 (95% CI 1.17 to 1.48) for those with educational level less than O level compared with those with O level qualifications or higher and 1.84 (95% CI 1.62 to 2.08) for high versus low area deprivation level. For women, the odds ratios for current smoking for manual social class were 1.14 (95% CI 1.03 to 1.27); 1.31 (95% CI 1.18 to 1.46) for low educational level and 1.68 (95% CI 1.49 to 1.90) for high residential deprivation respectively. CONCLUSIONS: Residential deprivation level using the Townsend score, individual social class, and educational level all independently predict smoking habit in both men and women. Efforts to reduce cigarette smoking need to tackle not just individual but also area based factors. Understanding the specific factors in deprived areas that influence smoking habit may help inform preventive efforts. PMID- 12646545 TI - No association of income inequality with adult mortality within New Zealand: a multi-level study of 1.4 million 25-64 year olds. AB - STUDY OBJECTIVE: To determine the association of regional income inequality within New Zealand with mortality among 25-64 year olds. DESIGN: Individual census and mortality records were linked over the 1991-94 period. Income inequality (Gini coefficients) and average household income variables were calculated for 35 regions. "Individual level" variables were sex, age, ethnicity, household income, rurality, and small area socioeconomic deprivation. Logistic regression was used for the analyses. Sensitivity analyses for the level of regional aggregation were conducted. PARTICIPANTS: 1.4 million New Zealand census respondents aged 25-64 years followed up for mortality for three years. MAIN RESULTS: Controlling for age, ethnicity, rurality, household income, and regional mean income, there was no association of income inequality with all cause mortality for either men (OR=1.007 for a 0.01 increase in the Gini, 95% confidence intervals 0.989 to 1.024) or women (OR=1.004, 0. 983 to 1.026). By cause of death (cancer, cardiovascular disease, unintentional injury, and suicide) there was some suggestion of a positive association for female unintentional injury (OR=1.068, 0.952 to 1.198) and suicide (OR=1.087, 0.957 to 1.234) but the 95% confidence intervals all included 1.0. Failure to control for ethnicity at the individual level resulted in some association of increasing regional income inequality with increasing mortality risk. Using fewer (n=14) or more (n=73) regional divisions did not substantially change the findings. CONCLUSION: There is no convincing evidence of an association of income inequality within New Zealand with adult mortality. Previous ecological analyses within New Zealand suggesting an association of income inequality with mortality were confounded by ethnicity at the individual level. However, this study does not refute the possibility that income inequality at the national level affects health. PMID- 12646546 TI - Do psychosocial work factors and social relations exert independent effects on sickness absence? A six year prospective study of the GAZEL cohort. AB - STUDY OBJECTIVES: The objective of this prospective cohort study was to determine whether psychosocial work characteristics and social relations exert independent effects on the incidence of sickness absence in a population of middle aged French employees over six years of follow up. DESIGN: This study included 9631 men and 3595 women participating in the French GAZEL cohort. Social relations (social networks, personal social support, and social relations satisfaction) were measured in 1994 by self report. Psychosocial work characteristics (decision latitude, psychological demands, and social support at work) were ascertained in 1995. Sickness absence data were collected independently. The authors studied the incidence of short (>7 days), intermediate (7-21 days), and long (>21 days) spells of absence from 1995 to 31 December 2001. Rate ratios associated with psychosocial exposures, adjusted on sociodemographic characteristics, and health behaviours, were calculated by means of log-linear Poisson regression. SETTING: A cohort of 20000 employees of France's national gas and electricity company (the GAZEL study). MAIN RESULTS: Among men and women, levels of decision latitude and personal social support below the median predicted 17% to 24% increases in absence rates. Low satisfaction with social relations and low social support at work lead to a 10% to 26% excess in sick leaves among men. No interactive effects were found between the variables under study. CONCLUSIONS: The quality of the work environment and of social relations affect sickness absence over an extended period of follow up. This study supports the hypothesis of independent, not interactive effects. PMID- 12646548 TI - Inequalities in survival from colorectal cancer: a comparison of the impact of deprivation, treatment, and host factors on observed and cause specific survival. AB - OBJECTIVE: To investigate whether socioeconomic deprivation is associated with cause specific and all cause survival for colorectal cancer and to what extent this is independent of significant prognostic factors. DESIGN: Prospective cohort. SETTING: The former Wessex Health Region, South West England. PARTICIPANTS: All patients resident in Wessex registered with a diagnosis of colorectal cancer over three years (n=5176). Survival analysis was carried out on those patients with compete data for all factors and a positive survival time (n=4419). OUTCOMES: Death from colorectal cancer and all cause over five year follow up from initial diagnosis. MAIN RESULTS: Deprivation was significantly associated with survival for both outcomes in univariate analysis; the unadjusted hazard ratio for dying from colorectal cancer (most deprived compared with most affluent) was 1.12 (95% CI 1.00 to 1.25) and for all cause was 1.18 (1.07 to 1.30). Significant prognostic factors for both outcomes were age, specialisation of surgeon, Dukes's stage, and emergency compared with elective surgery. Comorbidity and gender were only associated with all cause survival. After adjustment for prognostic factors, the effect of deprivation on both cause specific and all cause mortality was reduced, and it was non-significant for colorectal cancer. However, the most deprived group had consistently worse survival than the most affluent. CONCLUSIONS: Factors associated with survival with colorectal cancer depend on the outcome measure. Socioeconomic deprivation is adversely associated with survival in patients with colorectal cancer. This is strongest for non-colorectal cancer death, partly reflecting higher comorbidity, but it is there for colorectal cancer though not statistically significant. Conclusive evidence of the inequalities by socioeconomic status and underlying reasons needs to come from studies using individual based measures of socioeconomic status and more detail on treatment and host related factors. PMID- 12646549 TI - Retrospective cohort study examining incidence of HIV and hepatitis C infection among injecting drug users in Dublin. PMID- 12646547 TI - Health status of users of hormone replacement therapy by hysterectomy status in Western Australia. AB - STUDY OBJECTIVES: To compare the demographic, behavioural, and biological correlates of use of hormone replacement therapy (HRT) in women with an intact uterus and women who have undergone hysterectomy. DESIGN: Cross sectional analysis of data from the Busselton Health Study and the 1994 Healthway-National Heart Foundation Risk Factor Survey. SETTING: Busselton and Perth, Western Australia, 1994. PARTICIPANTS: 2540 women aged 35-79 years. MAIN OUTCOME MEASURES: Demographic, behavioural, and biological correlates of use of HRT by hysterectomy status. RESULTS: In women with an intact uterus, after adjustment for age and place of residence, current use of HRT was significantly associated with having a professional level of occupation, ever use of alcohol, having a history of smoking, and a lower body mass index. Current users of HRT had significantly lower levels of total cholesterol and higher levels of triglycerides than non-users. In women who had undergone hysterectomy, the only non-biological characteristic associated with use of HRT was having a history of smoking. Current users of HRT had lower levels of systolic blood pressure, lower levels of LDL cholesterol, higher levels of HDL cholesterol, and higher levels of triglycerides. The association between use of HRT and participation in exercise, level of systolic blood pressure, level of HDL cholesterol, and total/HDL cholesterol ratio varied significantly by hysterectomy status. After adjustment for age and place of residence, the mean levels of systolic and diastolic blood pressure, body mass index, waist/hip ratio, LDL cholesterol, and total/HDL cholesterol ratio were highest in women who had undergone hysterectomy and were not using HRT. CONCLUSIONS: Demographic/behavioural and biological correlates of use of HRT varied depending on hysterectomy status. Demographic and behavioural characteristics were more important as selection factors for use of HRT in women with an intact uterus than in women who had undergone hysterectomy. Women who had undergone hysterectomy and were not using HRT had a significantly worse profile for CHD than did women with an intact uterus. These results indicate that any bias in estimates of the protective effect of HRT on risk of CHD in observational studies is likely to depend on the prevalence of hysterectomy within the study population. Hysterectomy status needs to be taken into account in any studies that investigate the effect of HRT on risk of CHD. PMID- 12646550 TI - A superficial glance at child health. PMID- 12646551 TI - Health benefits of green spaces not confirmed. PMID- 12646552 TI - Using double-stranded RNA to prevent in vitro and in vivo viral infections by recombinant baculovirus. AB - Introduction of double-stranded RNA (dsRNA) into a wide variety of cells and organisms results in post-transcriptional depletion of the homologue endogenous mRNA. This well-preserved phenomenon known as RNA interference (RNAi) is present in evolutionarily diverse organisms such as plants, fungi, insects, metazoans, and mammals. Because the identification of the targeted mRNA by the RNAi machinery depends upon Watson-Crick base-pairing interactions, RNAi can be exquisitely specific. We took advantage of this powerful and flexible technique to demonstrate that selective silencing of genes essential for viral propagation prevents in vitro and in vivo viral infection. Using the baculovirus Autographa californica, a rapidly replicating and highly cytolytic double-stranded DNA virus that infects many different insect species, we show for the first time that introduction of dsRNA from gp64 and ie1, two genes essential for baculovirus propagation, results in prevention of viral infection in vitro and in vivo. This is the first report demonstrating the use of RNAi to inhibit a viral infection in animals. This inhibition was specific, because dsRNA from the polyhedrin promoter (used as control) or unrelated dsRNAs did not affect the time course of viral infection. The most relevant consequences from the present study are: 1) RNAi offers a rapid and efficient way to interfere with viral genes to assess the role of specific proteins in viral function and 2) using RNAi to interfere with viral genes essential for cell infection may provide a powerful therapeutic tool for the treatment of viral infections. PMID- 12646553 TI - Nitrosylation of cytochrome c during apoptosis. AB - Cytochrome c released from mitochondria into the cytoplasm plays a critical role in many forms of apoptosis by stimulating apoptosome formation and subsequent caspase activation. However, the mechanisms regulating cytochrome c apoptotic activity are not understood. Here we demonstrate that cytochrome c is nitrosylated on its heme iron during apoptosis. Nitrosylated cytochrome c is found predominantly in the cytoplasm in control cells. In contrast, when cytochrome c release from mitochondria is inhibited by overexpression of the anti apoptotic proteins B cell lymphoma/leukemia (Bcl)-2 or Bcl-X(L), nitrosylated cytochrome c is found in the mitochondria. These data suggest that during apoptosis, cytochrome c is nitrosylated in mitochondria and then rapidly released into the cytoplasm in the absence of Bcl-2 or Bcl-X(L) overexpression. In vitro nitrosylation of cytochrome c increases caspase-3 activation in cell lysates. Moreover, the inhibition of intracellular cytochrome c nitrosylation is associated with a decrease in apoptosis, suggesting that cytochrome c nitrosylation is a proapoptotic modification. We conclude that nitrosylation of the heme iron of cytochrome c may be a novel mechanism of apoptosis regulation. PMID- 12646554 TI - Histamine antagonizes tumor necrosis factor (TNF) signaling by stimulating TNF receptor shedding from the cell surface and Golgi storage pool. AB - Tumor necrosis factor (TNF) activates pro-inflammatory functions of vascular endothelial cells (EC) through binding to receptor type 1 (TNFR1) molecules expressed on the cell surface. The majority of TNFR1 molecules are localized to the Golgi apparatus. Soluble forms of TNFR1 (as well as of TNFR2) can be shed from the EC surface and inhibit TNF actions. The relationships among cell surface, Golgi-associated, and shed forms of TNFR1 are unclear. Here we report that histamine causes transient loss of surface TNFR1, TNFR1 shedding, and mobilization of TNFR1 molecules from the Golgi in cultured human EC. The Golgi pool of TNFR1 serves both to replenish cell surface receptors and as a source of shed receptor. Histamine-induced shedding is blocked by TNF-alpha protease inhibitor, an inhibitor of TNF-alpha-converting enzyme, and through the H1 receptor via a MEK-1/p42 and p44 mitogen-activated protein kinase pathway. Cultured EC with histamine-induced surface receptor loss become transiently refractory to TNF. Histamine injection into human skin engrafted on immunodeficient mice similarly caused shedding of TNFR1 and diminished TNF mediated induction of endothelial adhesion molecules. These results both clarify relationships among TNFR1 populations and reveal a novel anti-inflammatory activity of histamine. PMID- 12646555 TI - C-terminal octylation rescues an inactive T20 mutant: implications for the mechanism of HIV/SIMIAN immunodeficiency virus-induced membrane fusion. AB - T20, a synthetic peptide corresponding to a C-terminal segment of the envelope glycoprotein (gp41) of human and simian immunodeficiency viruses, is a potent inhibitor of viral infection. We report here that C-terminal octylation of simian immunodeficiency virus gp41-derived T20 induces a significant increase in its inhibitory potency. Furthermore, when C-terminally octylated, an otherwise inactive mutant in which the C-terminal residues GNWF were replaced by ANAA has potency similar to that of the wild type T20. This effect cannot be explained by a trivial inhibitory effect of the octyl group added to the peptides, because the N-terminally octylated peptides have the same activity as the non-octylated parent peptides. The effects caused by octylation on the oligomerization, secondary structure, and membrane-interaction properties of the peptides were investigated. Our results shed light on the mechanism of inhibition by T20 and provide experimental support for the existence of a pre-hairpin intermediate. PMID- 12646556 TI - Regulation of dopamine D1 receptor trafficking and desensitization by oligomerization with glutamate N-methyl-D-aspartate receptors. AB - Activation of dopamine D1 receptors is critical for the generation of glutamate induced long-term potentiation at corticostriatal synapses. In this study, we report that, in striatal neurons, D1 receptors are co-localized with N-methyl-d aspartate (NMDA) receptors in the postsynaptic density and that they co immunoprecipitate with NMDA receptor subunits from postsynaptic density preparations. Using modified bioluminescence resonance energy transfer, we demonstrate that D1 and NMDA receptor clustering reflects the existence of direct interactions. The tagged D1 receptor and NR1 subunit cotransfected in COS-7 cells generated a significant bioluminescence resonance energy transfer signal that was insensitive to agonist stimulation and that did not change in the presence of the NR2B subunit, suggesting that the D1 receptor constitutively and selectively interacts with the NR1 subunit of the NMDA channel. Oligomerization with the NR1 subunit substantially modified D1 receptor trafficking. In individually transfected HEK293 cells, NR1 was localized in the endoplasmic reticulum, whereas the D1 receptor was targeted to the plasma membrane. In cotransfected cells, both the D1 receptor and NR1 subunit were retained in cytoplasmic compartments. In the presence of the NR2B subunit, the NR1-D1 receptor complex was translocated to the plasma membrane. These data suggest that D1 and NMDA receptors are assembled within intracellular compartments as constitutive heteromeric complexes that are delivered to functional sites. Coexpression with NR1 and NR2B subunits also abolished agonist-induced D1 receptor cytoplasmic sequestration, indicating that oligomerization with the NMDA receptor could represent a novel regulatory mechanism modulating D1 receptor desensitization and cellular trafficking. PMID- 12646557 TI - Two aromatic residues in the PB2 subunit of influenza A RNA polymerase are crucial for cap binding. AB - mRNAs are capped at their 5'-end by a unique cap structure containing N7-methyl guanine. Recognition of the cap structure is of paramount importance in some of the most central processes of gene expression as well as in some viral processes, such as priming of influenza virus transcription. The recent resolution of the structure of three evolutionary unrelated cap binding proteins, the vaccinia viral protein VP39, the eukaryotic translation factor eIF4E, and the nuclear cap binding protein CBP20 showed that the recognition of the cap structure is achieved by the same general mechanism, i.e. by "sandwiching" of the N7-methyl guanine of the cap structure between two aromatic amino acid residues. The purpose of the present study was to test whether a similar cap recognition mechanism had independently evolved for the RNA polymerase of influenza virus. Combining in vivo and in vitro methods, we characterized two crucial aromatic amino acids, Phe363 and Phe404, in the PB2 subunit of the viral RNA polymerase that are essential for cap binding. The aromaticity of these two residues is conserved in influenza A, B, and C and even in the divergent Thogoto virus PB2 subunits. Thus, our results favor a similar mechanism of cap binding by the influenza RNA polymerase as in the evolutionary unrelated VP39, eIF4E, and CBP20. PMID- 12646558 TI - A2E-epoxides damage DNA in retinal pigment epithelial cells. Vitamin E and other antioxidants inhibit A2E-epoxide formation. AB - The autofluorescent pigments that accumulate in retinal pigment epithelial cells with aging and in some retinal disorders have been implicated in the etiology of macular degeneration. The major constituent is the fluorophore A2E, a pyridinium bisretinoid. Light-exposed A2E-laden retinal pigment epithelium exhibits a propensity for apoptosis with light in the blue region of the spectrum being most damaging. Efforts to understand the events precipitating the death of the cells have revealed that during irradiation (430 nm), A2E self-generates singlet oxygen with the singlet oxygen in turn reacting with A2E to generate epoxides at carbon carbon double bonds. Here we demonstrate that A2E-epoxides, independent of singlet oxygen, exhibit reactivity toward DNA with oxidative base changes being at least one of these lesions. Mass spectrometry revealed that the antioxidants vitamins E and C, butylated hydroxytoluene, resveratrol, a trolox analogue (PNU 83836-E), and bilberry extract reduce A2E-epoxidation, whereas single cell gel electrophoresis and cell viability studies revealed a corresponding reduction in the incidence of DNA damage and cell death. Vitamin E, a lipophilic antioxidant, produced a more pronounced decrease in A2E-epoxidation than vitamin C, and treatment with both vitamins simultaneously did not confer additional benefit. Studies in which singlet oxygen was generated by endoperoxide in the presence of A2E revealed that vitamin E, butylated hydroxytoluene, resveratrol, the trolox analogue, and bilberry reduced A2E-epoxidation by quenching singlet oxygen. Conversely, vitamin C and ginkgolide B were not efficient quenchers of singlet oxygen under these conditions. PMID- 12646559 TI - Stress-induced protein phosphatase 2C is a negative regulator of a mitogen activated protein kinase. AB - Protein phosphatases of type 2C (PP2Cs) play important roles in eukaryotic signal transduction. In contrast to other eukaryotes, plants such as Arabidopsis have an unusually large group of 69 different PP2C genes. At present, little is known about the functions and substrates of plant PP2Cs. We have previously shown that MP2C, a wound-induced alfalfa PP2C, is a negative regulator of mitogen-activated protein kinase (MAPK) pathways in yeast and plants. In this report, we provide evidence that alfalfa salt stress-inducible MAPK (SIMK) and stress-activated MAPK (SAMK) are activated by wounding and that MP2C is a MAPK phosphatase that directly inactivates SIMK but not the wound-activated MAPK, SAMK. SIMK is inactivated through threonine dephosphorylation of the pTEpY motif, which is essential for MAPK activity. Mutant analysis indicated that inactivation of SIMK depends on the catalytic activity of MP2C. A comparison of MP2C with two other PP2Cs, ABI2 and AtP2CHA, revealed that although all three phosphatases have similar activities toward casein as a substrate, only MP2C is able to dephosphorylate and inactivate SIMK. In agreement with the notion that MP2C interacts directly with SIMK, the MAPK was identified as an interacting partner of MP2C in a yeast two-hybrid screen. MP2C can be immunoprecipitated with SIMK in a complex in vivo and shows direct binding to SIMK in vitro in protein interaction assays. Wound-induced MP2C expression correlates with the time window when SIMK is inactivated, corroborating the notion that MP2C is involved in resetting the SIMK signaling pathway. PMID- 12646560 TI - Activation of the ERK1/2 signaling pathway promotes phosphorylation and proteasome-dependent degradation of the BH3-only protein, Bim. AB - Both the ERK and phosphatidylinositol 3'-kinase (PI3K) signaling pathways can protect cells from apoptosis following withdrawal of survival factors. We have previously shown that the ERK1/2 pathway acts independently of PI3K to block expression of the BH3-only protein, BimEL, and prevent serum withdrawal-induced cell death, although the precise mechanism by which ERK reduced BimEL levels was unclear. By comparing Bim mRNA and Bim protein, expression we now show that the rapid expression of BimEL following serum withdrawal cannot be accounted for simply by increases in mRNA following inhibition of PI3K. In cells maintained in serum BimEL is a phosphoprotein. We show that activation of the ERK1/2 pathway is both necessary and sufficient to promote BimEL phosphorylation and that this leads to a substantial increase in turnover of the BimEL protein. ERK1/2 dependent degradation of BimEL proceeds via the proteasome pathway because it is blocked by proteasome inhibitors and is defective at the restrictive temperature in cells with a temperature-sensitive mutation in the E1 component of the ubiquitin-conjugating system. Finally, co-transfection of BimEL and FLAG ubiquitin causes the accumulation of polyubiquitinated forms of Bim, and this requires the ERK1/2 pathway. Our findings provide new insights into the regulation of Bim and the role of the ERK pathway in cell survival. PMID- 12646561 TI - Enhancer-dependent 5'-splice site control of fruitless pre-mRNA splicing. AB - The Drosophila fruitless (fru) gene encodes a transcription factor that essentially regulates all aspects of male courtship behavior. The use of alternative 5'-splice sites generates fru isoforms that determine gender appropriate sexual behaviors. Alternative splicing of fru is regulated by TRA and TRA2 and depends on an exonic splicing enhancer (fruRE) consisting of three 13 nucleotide repeat elements, nearly identical to those that regulate alternative sex-specific 3'-splice site choice in the doublesex (dsx) gene. dsx has provided a useful model system to investigate the mechanisms of enhancer-dependent 3' splice site choice. However, little is known about enhancer-dependent regulation of alternative 5'-splice sites. The mechanisms of this process were investigated using an in vitro system in which recombinant TRA/TRA2 could activate the female specific 5'-splice site of fru. Mutational analysis demonstrated that one 13 nucleotide repeat element within the fruRE is required and sufficient to activate the regulated female-specific splice site. As was established for dsx, the fruRE can be replaced by a short element encompassing tandem 13-nucleotide repeat elements, by heterologous splicing enhancers, and by artificially tethering a splicing activator to the pre-mRNA. Complementation experiments showed that Ser/Arg-rich proteins facilitate enhancer-dependent 5'-splice site activation. We conclude that splicing enhancers function similarly in activating regulated 5'- and 3'-splice sites. These results suggest that exonic splicing enhancers recruit multiple spliceosomal components required for the initial recognition of 5'- and 3'-splice sites. PMID- 12646562 TI - Transcription arrest at a lesion in the transcribed DNA strand in vitro is not affected by a nearby lesion in the opposite strand. AB - Cis-syn cyclobutane pyrimidine dimers (CPDs) are the most frequently formed lesions in UV-irradiated DNA. CPDs are repaired by the nucleotide excision repair pathway. Additionally, they are subject to transcription-coupled DNA repair. In the general model for transcription-coupled DNA repair, an RNA polymerase arrested at a lesion on the transcribed DNA strand facilitates repair by recruiting the repair machinery to the site of the lesion. Consistent with this model, transcription experiments in vitro have shown that CPDs in the transcribed DNA strand interfere with the translocation of prokaryotic and eukaryotic RNA polymerases. Here, we study the behavior of RNA polymerase when transcribing a template that contains two closely spaced lesions, one on each DNA strand. Similar DNA templates containing no CPD, or a single CPD on either the transcribed or the nontranscribed strand were used as controls. Using an in vitro transcription system with purified T7 RNA polymerase (T7 RNAP) or rat liver RNAP II, we characterized transcript length and efficiency of transcription in vitro. We also tested the sensitivity of the arrested RNAP II-DNA-RNA ternary complex, at a CPD in the transcribed strand, to transcription factor TFIIS. The presence of a nearby CPD in the nontranscribed strand did not affect the behavior of either RNA polymerase nor did it affect the reverse translocation ability of the RNAP II-arrested complex. Our results additionally indicate that the sequence context of a CPD affects the efficiency of T7 RNAP arrest more significantly than that of RNAP II. PMID- 12646563 TI - Rrn3 becomes inactivated in the process of ribosomal DNA transcription. AB - The human homologue of yeast Rrn3, a 72-kDa protein, is essential for ribosomal DNA (rDNA) transcription. Although the importance of Rrn3 function in rDNA transcription is well established, its mechanism of action has not been determined. It has been suggested that the phosphorylation of either yeast RNA polymerase I or mammalian Rrn3 regulates the formation of RNA polymerase I.Rrn3 complexes that can interact with the committed template. These and other reported differences would have implications with respect to the mechanism by which Rrn3 functions in transcription. For example, in the yeast rDNA transcription system, Rrn3 might function catalytically, but in the mammalian system it might function stoichiometrically. Thus, we examined the question as to whether Rrn3 functions catalytically or stoichiometrically. We report that mammalian Rrn3 becomes the limiting factor as transcription reactions proceed. Moreover, we demonstrate that Rrn3 is inactivated during the transcription reactions. For example, Rrn3 isolated from a reaction that had undergone transcription cannot activate transcription in a subsequent reaction. We also show that this inactivated Rrn3 not only dissociates from RNA polymerase I, but is not capable of forming a stable complex with RNA polymerase I. Our results indicate that Rrn3 functions stoichiometrically in rDNA transcription and that its ability to associate with RNA polymerase I is lost upon transcription. PMID- 12646564 TI - Increased constitutive c-Jun N-terminal kinase signaling in mice lacking glutathione S-transferase Pi. AB - Glutathione S-transferase Pi (GSTP) detoxifies electrophiles by catalyzing their conjugation with reduced glutathione. A second function of this protein in cell defense has recently been proposed that is related to its ability to interact with c-Jun N-terminal kinase (JNK). The present study aimed to determine whether this interaction results in increased constitutive JNK activity in the absence of GSTP in GstP1/P2(-/-) mice and whether such a phenomenon leads to the up regulation of genes that are relevant to cell defense. We found a significant increase in constitutive JNK activity in the liver and lung of GstP1/P2-/- compared with GstP1/P2(+/+) mice. The greatest increase in constitutive JNK activity was observed in null liver and was accompanied by a significant increase in activator protein-1 DNA binding activity (8-fold) and in the mRNA levels for the antioxidant protein heme oxygenase-1 compared with wild type. Furthermore UDP glucuronosyltransferase 1A6 mRNA levels were significantly higher in the livers of GstP1/P2(-/-) compared with GstP1/P2(+/+) mice, which correlated to a 2-fold increase in constitutive activity both in vitro and in vivo. There was no difference in the gene expression of other UDP-glucuronosyltransferase isoforms, manganese superoxide dismutase, microsomal epoxide hydrolase, or GSTA1 between GstP1/P2(-/-) and GstP1/P2(+/+) mice. Additionally there was no phenotypic difference in the induction of heme oxygenase-1 mRNA after acetaminophen administration. This study not only demonstrates the role of GSTP as a direct inhibitor of JNK in vivo but also its role in regulating the constitutive expression of specific downstream molecular targets of the JNK signaling pathway. PMID- 12646565 TI - Synergistic assembly of linker for activation of T cells signaling protein complexes in T cell plasma membrane domains. AB - Transmembrane adaptor molecule LAT (linker for activation of T cells) forms a central scaffold for signaling protein complexes that accumulate in the vicinity of activated T cell antigen receptors (TCR). Here we used biochemical analysis of immunoisolated plasma membrane domains and fluorescence imaging of green fluorescence protein-tagged signaling proteins to investigate the contributions of different tyrosine-based signaling protein docking sites of LAT to the formation of LAT signaling protein assemblies in TCR membrane domains. We found that the phospholipase C gamma docking site of LAT and different Grb2/Gads docking sites function in an interdependent fashion and synergize to accumulate LAT, Grb2, and phospholipase C gamma in TCR signaling assemblies. Two-dimensional gels showed that Grb2 is a predominant cytoplasmic adaptor in the isolated LAT signaling complexes, whereas Gads, Crk-1, and Grap are present in lower amounts. Taken together our data suggest a synergistic assembly of multimolecular TCR.LAT signal transduction complexes in T cell plasma membrane domains. PMID- 12646566 TI - Nephrin and Neph1 co-localize at the podocyte foot process intercellular junction and form cis hetero-oligomers. AB - Glomerular visceral epithelial cells (podocytes) appear to play a central role in maintaining the selective filtration barrier of the renal glomerulus. While the immunoglobulin superfamily member Nephrin was proposed to act as a cell adhesion molecule at the podocyte intercellular junction necessary for maintaining glomerular perm selectivity, the Nephrin ligand has not been identified. The existence of a new subfamily of Nephrin-like molecules including Neph1 was recently described. Genetic deletion of Nephrin or Neph1 resulted in similar phenotypes of podocyte foot process effacement and proteinuria. The subcellular localization of Neph1 and the possibility that Nephrin and Neph1 interact was investigated. Polyclonal antiserum for Neph1 was raised and characterized. Neph1 migrated as a 90-kDa protein on SDS-PAGE under reducing conditions. Neph1 was identified in a glomerular and podocyte-specific distribution in adult rat kidney. Like Nephrin and Podocin, Neph1 was enriched in Triton X-100 detergent resistant membrane fractions. Consistent with this observation, immunogold electron microscopy demonstrated that Neph1 localized exclusively to lateral margins of podocyte foot processes at the insertion of the slit diaphragm. Neph1 and Nephrin participate in a direct cis-interaction involving their cytoplasmic domains. In addition, interactions between the extracellular domain of Nephrin and itself and between the extracellular domain of Nephrin and that of Neph1 were detected. Neph1 did not interact via a homophilic interaction. These observations suggest that Nephrin and Neph1 form a hetero-oligomeric receptor complex in the plane of the membrane that might interact across the foot process intercellular junction through interactions between Nephrin with itself and Neph1. PMID- 12646567 TI - Mammalian fibroblasts lacking mitochondrial NAD+-dependent methylenetetrahydrofolate dehydrogenase-cyclohydrolase are glycine auxotrophs. AB - Primary fibroblasts established from embryos of NAD-dependent mitochondrial methylenetetrahydrofolate dehydrogenase-cyclohydrolase (NMDMC) knockout mice were spontaneously immortalized or transformed with SV40 Large T antigen. Mitotracker Red CMXRos staining of the cells indicates the presence of intact mitochondria with a membrane potential. The nmdmc(-/-) cells are auxotrophic for glycine, demonstrating that NMDMC is the only methylenetetrahydrofolate dehydrogenase normally expressed in the mitochondria of these cell lines. Growth of null mutant but not wild type cells on complete medium with dialyzed serum is stimulated about 2-fold by added formate or hypoxanthine. Radiolabeling experiments demonstrated a 3-10 x enhanced incorporation of radioactivity into DNA from formate relative to serine by nmdmc(-/-) cells. The generation of one-carbon units by mitochondria in nmdmc(-/-) cells is completely blocked, and the cytoplasmic folate pathways alone are insufficient for optimal purine synthesis. The results demonstrate a metabolic role for NMDMC in supporting purine biosynthesis. Despite the recognition of these metabolic defects in the mutant cell lines, the phenotype of nmdmc(-/-) embryos that begin to die at E13.5 is not improved when pregnant dams are given a glycine-rich diet or daily injections of sodium formate. PMID- 12646568 TI - Distinct mechanisms for regulating the tumor suppressor and antiapoptotic functions of Rb. AB - The retinoblastoma protein, Rb, suppresses tumorigenesis by inhibiting cell proliferation and promoting senescence and differentiation. Paradoxically, Rb also inhibits apoptosis, which would seem to oppose its tumor suppressor function. Further, most human cancer cells inactivate Rb by hyperphosphorylation and demonstrate increased proliferative capacity but not high levels of apoptosis. As a potential explanation for these findings, we show here that the tumor suppressor and antiapoptotic functions of Rb are regulated by distinct phosphorylation events. Phosphorylation of sites in the C terminus occurs efficiently every cell cycle and regulates proliferation. Phosphorylation of Ser567 is inefficient and does not occur during the normal cell cycle. However, high cyclin-dependent kinase activity promotes phosphorylation of Ser567 by inducing an intramolecular interaction that leads to release of E2F, degradation of Rb, and susceptibility to apoptosis. Thus, phosphorylation of Ser567 may limit excessive proliferation by triggering cell death under hyperproliferative conditions. These findings suggest that the antiproliferative and antiapoptotic activities of Rb may represent complementary functions that work in concert to maintain the proliferation rate of cells within certain limits. As a survival strategy, some cancer cells may exploit this dual role of Rb by phosphorylating sites that regulate tumor suppression but avoiding phosphorylation of Ser567 and consequent apoptotic stimulus. PMID- 12646569 TI - Phosphorylation enhances mitochondrial targeting of GSTA4-4 through increased affinity for binding to cytoplasmic Hsp70. AB - Recently we showed that three different isoforms of cytosolic glutathione S transferases (GST), including GSTA4-4, are also localized in the mitochondrial compartment. In this study, we have investigated the mechanism of mouse GSTA4-4 targeting to mitochondria, using a combination of in vitro mitochondrial import assay and in vivo targeting in COS cells transfected with cDNA. Our results show that the mitochondrial GSTA4-4 is more heavily phosphorylated compared with its cytosolic counterpart. Protein kinase activators (cAMP, forskolin, or phorbol-12 myristate-13-acetate) markedly increased GSTA4-4 targeting to mitochondria, whereas kinase inhibitors caused its retention in the cytosol. Immunoinhibition and immunodepletion studies showed that the Hsp70 chaperone is required for the efficient translation of GSTA4-4 as well as its translocation to mitochondria. Co immunoprecipitation studies showed that kinase inhibitors attenuate the affinity of GSTA4-4 for cytoplasmic Hsp70 suggesting the importance of phosphorylation for binding to the chaperone. Mutational analysis show that the putative mitochondrial targeting signal resides within the C-terminal 20 amino acid residues of the protein and that the targeting signal requires activation by phosphorylation at the C-terminal-most protein kinase A (PKA) site at Ser-189 or protein kinase C (PKC) site at Thr-193. We demonstrate for the first time that PKA and PKC modulate the cytoplasmic and mitochondrial pools of GSTA4-4. PMID- 12646570 TI - Calreticulin is at the surface of circulating neutrophils and uses CD59 as an adaptor molecule. AB - Calreticulin, which has been proposed to be a C1q receptor on neutrophils, has neither a transmembrane domain nor a GPI-anchor attachment site and must utilize an adaptor molecule to attach to the plasma membrane. The expression of ecto calreticulin on purified human neutrophils did not result from contamination by soluble or intracellular calreticulin released during cell fractionation because it was expressed on circulating neutrophils, and the expression did not increase significantly with neutrophil isolation. All neutrophils expressed calreticulin with a unimodal distribution. Treatment of neutrophils with either a cholesterol binding agent or phosphatidylinositol-specific phospholipase C dramatically decreased ecto-calreticulin expression indicating that the adaptor molecule(s) are located in lipid rafts and have a GPI-anchor. Analysis for the co-expression of specific GPI-anchored proteins and ecto-calreticulin in cells that were deficient in specific GPI-anchored proteins, indicated that ecto-calreticulin was best associated with CD59. Calreticulin reciprocally immunoprecipited with CD59, which provided direct evidence that CD59 is an adaptor for ecto-calreticulin. Immunofluorescence and confocal microscopy demonstrated that ecto-calreticulin co localized with a fraction of CD59 at the cell surface. Cross-linking ecto calreticulin with antibodies induced a Ca2+ flux, which suggests that ecto calreticulin is capable of signaling following ligand binding. Ecto-calreticulin has been associated with diverse cellular functions. An appreciation that the adaptors for ecto-calreticulin are GPI-anchored will provide a framework for understanding any common features underlying ecto-calreticulin ligation. PMID- 12646571 TI - A fast-acting, modular-structured staphylokinase fusion with Kringle-1 from human plasminogen as the fibrin-targeting domain offers improved clot lysis efficacy. AB - To develop a fast-acting clot dissolving agent, a clot-targeting domain derived from the Kringle-1 domain in human plasminogen was fused to the C-terminal end of staphylokinase with a linker sequence in between. Production of this fusion protein in Bacillus subtilis and Pichia pastoris was examined. The Kringle domain in the fusion protein produced from B. subtilis was improperly folded because of its complicated disulfide-bond profile, whereas the staphylokinase domain produced from P. pastoris was only partially active because of an N-linked glycosylation. A change of the glycosylation residue, Thr-30, to alanine resulted in a non-glycosylated biologically active fusion. The resulting mutein, designated SAKM3-L-K1, was overproduced in P. pastoris. Each domain in SAKM3-L-K1 was functional, and this fusion showed fibrin binding ability by binding directly to plasmin-digested clots. In vitro fibrin clot lysis in a static environment and plasma clot lysis in a flow-cell system demonstrated that the engineered fusion outperformed the non-fused staphylokinase. The time required for 50% clot lysis was reduced by 20 to 500% under different conditions. Faster clot lysis can potentially reduce the degree of damage to occluded heart tissues. PMID- 12646572 TI - Direct observation of amyloid fibril growth monitored by thioflavin T fluorescence. AB - Real-time monitoring of fibril growth is essential to clarify the mechanism of amyloid fibril formation. Thioflavin T (ThT) is a reagent known to become strongly fluorescent upon binding to amyloid fibrils. Here, we show that, by monitoring ThT fluorescence with total internal reflection fluorescence microscopy (TIRFM), amyloid fibrils of beta2-microgobulin (beta2-m) can be visualized without requiring covalent fluorescence labeling. One of the advantages of TIRFM would be that we selectively monitor fibrils lying along the slide glass, so that we can obtain the exact length of fibrils. This method was used to follow the kinetics of seed-dependent beta2-m fibril extension. The extension was unidirectional with various rates, suggesting the heterogeneity of the amyloid structures. Since ThT binding is common to all amyloid fibrils, the present method will have general applicability for the analysis of amyloid fibrils. We confirmed this with the octapeptide corresponding to the C terminus derived from human medin and the Alzheimer's amyloid beta-peptide. PMID- 12646573 TI - Presenilin 1 and presenilin 2 have differential effects on the stability and maturation of nicastrin in Mammalian brain. AB - The presenilins and nicastrin form high molecular mass, multimeric protein complexes involved in the intramembranous proteolysis of several proteins. Post translational glycosylation and trafficking of nicastrin is necessary for the activity of these complexes. We report here that although there are differences in the post-translational processing of nicastrin in neurons and glia, both of the presenilins are required for the physiological post-translational modification and for the correct subcellular distribution of nicastrin. Absence of presenilin 1 (PS1) is associated with dramatic reductions in the level of mature glycosylated nicastrin and with redistribution of nicastrin away from the cell surface. In contrast, absence of presenilin 2 (PS2) is associated with only modest reductions in the levels of immature nicastrin. It is notable that these differential effects parallel the differential effects of null mutations in PS1 and PS2 on APP and Notch processing. Our data therefore suggest that the differential interactions of PS1 and PS2 with nicastrin reflect different functions for the PS1 and PS2 complexes. PMID- 12646574 TI - Matrix-independent activation of phosphatidylinositol 3-kinase, Stat3, and cyclin A-associated Cdk2 Is essential for anchorage-independent growth of v-Ros transformed chicken embryo fibroblasts. AB - The question remains open whether the signaling pathways shown to be important for growth and transformation in adherent cultures proceed similarly and play similar roles for cells grown under anchorage-independent conditions. Chicken embryo fibroblasts (CEF) infected with the avian sarcoma virus UR2, encoding the oncogenic receptor protein-tyrosine kinase (RPTK) v-Ros, or with two of its transformation-impaired mutants were grown in nonadherent conditions in methylcellulose (MC)-containing medium, and the signaling functions essential for Ros-induced anchorage-independent growth were analyzed. We found that the overall tyrosine phosphorylation of cellular proteins in CEF transformed by v-Ros or by two oncogenic nonreceptor protein-tyrosine kinases (PTKs), v-Src and v-Yes, was dramatically reduced in nonadherent conditions compared with that in adherent conditions, indicating that cell adhesion to the extracellular matrix plays an important role in efficient substrate phosphorylation by these constitutively activated PTKs. The UR2 transformation-defective mutants were differentially impaired compared with UR2 in the activation of phosphatidylinositol 3-kinase (PI 3-kinase) and Stat3 in nonadherent conditions. Consistently, the constitutively activated mutants of PI 3-kinase and Stat3 rescued the ability of the UR2 mutants to promote anchorage-independent growth. Conversely, dominant negative mutants of PI 3-kinase and Stat3 inhibited UR2-induced anchorage-independent growth. UR2 infected CEF grown in nonadherent conditions displayed faster cell cycle progression than the control or the UR2 mutant-infected cells, and this appeared to correlate with a PI 3-kinase-dependent increase in cyclin A-associated Cdk2 activity. Treatment of UR2-infected cells with Cdk2 inhibitors led to the loss of the anchorage-independent growth-promoting activity of UR2. In conclusion, we have adopted an experimental system enabling us to study the signaling pathways in cells grown under anchorage-independent conditions and have identified matrix independent activation of PI 3-kinase and Stat3 signaling functions, as well as the PI 3-kinase-dependent increase of cyclin A-associated Cdk2 kinase activity, to be critical for the Ros-PTK-induced anchorage-independent growth. PMID- 12646575 TI - The carboxyl terminus of human cytomegalovirus-encoded 7 transmembrane receptor US28 camouflages agonism by mediating constitutive endocytosis. AB - US28 is one of four 7 transmembrane (7TM) chemokine receptors encoded by human cytomegalovirus and has been shown to both signal and endocytose in a ligand independent, constitutively active manner. Here we show that the constitutive activity and constitutive endocytosis properties of US28 are separable entities in this viral chemokine receptor. We generated chimeric and mutant US28 proteins that were altered in either their constitutive endocytic (US28 Delta 300, US28 Delta 317, US28-NK1-ctail, and US28-ORF74-ctail) or signaling properties (US28R129A). By using this series of mutants, we show that the cytoplasmic tail domain of US28 per se regulates receptor endocytosis, independent of the signaling ability of the core domain of US28. The constitutive endocytic property of the US28 c-tail was transposable to other 7TM receptors, the herpes virus 8 encoded ORF74 and the tachykinin NK1 receptor (ORF74-US28-ctail and NK1-US28 ctail). Deletion of the US28 C terminus resulted in reduced constitutive endocytosis and consequently enhanced signaling capacity of all receptors tested as assessed by inositol phosphate turnover, NF-kappa B, and cAMP-responsive element-binding protein transcription assays. We further show that the constitutive endocytic property of US28 affects the action of its chemokine ligand fractalkine/CX3CL1 and show that in the absence of the US28 C terminus, fractalkine/CX3CL1 acts as an agonist on US28. This demonstrates for the first time that the endocytic properties of a 7TM receptor can camouflage the agonist properties of a ligand. PMID- 12646576 TI - ATP22, a nuclear gene required for expression of the F0 sector of mitochondrial ATPase in Saccharomyces cerevisiae. AB - Expression of the mitochondrial proton-translocating ATPase of Saccharomyces cerevisiae has been shown to depend on chaperones that target the F1 and F0 sectors of this inner membrane complex. Here we report a new gene, designated ATP22 (reading frame YDR350C on chromosome IV), that provides an essential function in the assembly of F0. ATP22 was cloned by transformation of C208/L2, a strain previously assigned to complementation group G99 of a collection of respiration-defective nuclear pet mutants. C208/L2 and the other atp22 mutants have oligomycin-insensitive F1-ATPase, suggesting that the lesion is confined to F0. This is supported by the sedimentation properties of the mutant ATPase and results of immunochemical analysis of F0 subunit polypeptides. Northern analysis of ATPase transcripts and in vivo pulse labeling of the mitochondrial translation products in the mutant indicate normal expression of subunits 6, 8, and 9, the three mitochondrial gene products of F0. Atp22p therefore functions at a post translational stage in assembly of F0. Localization studies indicate Atp22p to be a component of the mitochondrial inner membrane. Protease protection experiments further indicate that Atp22p faces the matrix side of the membrane where most of the ATPase proteins are located and assembled. PMID- 12646577 TI - Incadronate amplifies prostaglandin F2 alpha-induced vascular endothelial growth factor synthesis in osteoblasts. Enhancement of MAPK activity. AB - We have previously reported that prostaglandin F2 alpha (PGF2 alpha) activates p44/p42 mitogen-activated protein kinase (MAPK) through protein kinase C (PKC) in osteoblast-like MC3T3-E1 cells. In the present study, we investigated the mechanism of vascular endothelial growth factor (VEGF) synthesis induced by PGF2 alpha and the effect of incadronate on the VEGF synthesis in these cells. PGF2 alpha significantly stimulated the VEGF synthesis in a dose-dependent manner between 1 pm and 10 microm. Cycloheximide reduced the PGF2 alpha effect. PGF2 alpha increased the levels of mRNA for VEGF. Cloprostenol, a PGF2 alpha-sensitive receptor agonist, potently induced the VEGF synthesis. Indomethacin, an inhibitor of cyclooxygenase, significantly reduced the PGF2 alpha-induced VEGF synthesis. Bisindolylmaleimide, an inhibitor of PKC, reduced the PGF2 alpha-induced VEGF synthesis. The VEGF synthesis induced by PGF2 alpha was significantly attenuated in the PKC down-regulated cells. PGF2 alpha elicited the translocation of PKC beta I from cytosol to membrane fraction. PD98059 or U0126, inhibitors of MEK, suppressed the VEGF synthesis induced by PGF2 alpha. Farnesyltransferase inhibitor failed to affect the PGF2 alpha-induced VEGF synthesis. Incadronate enhanced the synthesis of VEGF induced by PGF2 alpha. NaF-induced VEGF synthesis was also amplified by incadronate. PD98059 suppressed the enhancement by incadronate of PGF2 alpha-induced VEGF synthesis. Incadronate markedly enhanced the phosphorylation of Raf-1, MEK1/2, and p44/p42 MAPK induced by PGF2 alpha or 12-O-tetradecanoylphorbol-13-acetate, a PKC activator. Incadronate significantly enhanced the cloprostenol-increased level of VEGF concentration in mouse plasma in vivo. These results strongly suggest that PGF2 alpha stimulates VEGF synthesis through the PKC-dependent activation of p44/p42 MAPK in osteoblasts and that the incadronate enhances the VEGF synthesis at the point between PKC and Raf-1. PMID- 12646578 TI - A novel lytic peptide composed of DL-amino acids selectively kills cancer cells in culture and in mice. AB - The high toxicity of most chemotherapeutic drugs and their inactivation by multidrug resistance phenotypes motivated extensive search for drugs with new modes of action. We designed a short cationic diastereomeric peptide composed of d- and l-leucines, lysines, and arginines that has selective toxicity toward cancer cells and significantly inhibits lung metastasis formation in mice (86%) with no detectable side effects. Its ability to depolarize the transmembrane potential of cancer cells at the same rate (within minutes) and concentration (3 micro m), at which it shows biological activity, suggests a killing mechanism that involves plasma membrane perturbation. Confocal microscopy experiments verified that the cells died as a result of acute injury, swelling, and bursting, suggesting necrosis. Biosensor binding experiments and attenuated total reflectance-Fourier transform infrared spectroscopy using model membranes have substantiated its high selectivity toward cancer cells. Although this is an initial study that looked at tumor formation rather than the ability of the peptides to reduce established tumors, the simple sequence of the peptide, its high solubility, substantial resistance to degradation, and inactivation by serum components might make it a good candidate for future anticancer treatment. PMID- 12646579 TI - Transforming growth factor-beta induces secretion of activated ADAMTS-2. A procollagen III N-proteinase. AB - The metalloproteinase ADAMTS-2 has procollagen I N-proteinase activity capable of cleaving procollagens I and II N-propeptides in vitro, whereas mutations in the ADAMTS-2 gene in dermatosparaxis and Ehlers-Danlos syndrome VIIC show this enzyme to be responsible in vivo for most biosynthetic processing of procollagen I N propeptides in skin. Yet despite its important role in the regulation of collagen deposition, information regarding regulation and substrate specificity of ADAMTS 2 has remained sparse. Here we demonstrate that ADAMTS-2 can, like the procollagen C-proteinases, be regulated by transforming growth factor-beta 1 (TGF beta 1), with implications for mechanisms whereby this growth factor effects net increases in formation of extracellular matrix. TGF-beta 1 induced ADAMTS-2 mRNA approximately 8-fold in MG-63 osteosarcoma cells in a dose- and time-dependent, cycloheximide-inhibitable manner, which appeared to operate at the transcriptional level. Secreted ADAMTS-2 protein induced by TGF-beta 1 was 132 kDa and was identical in size to the fully processed, active form of the protease. Biosynthetic processing of ADAMTS-2 to yield the 132-kDa form is shown to be a two-step process involving sequential cleavage by furin-like convertases at two sites. Surprisingly, purified recombinant ADAMTS-2 is shown to cleave procollagen III N-propeptides as effectively as those of procollagens I and II, whereas processing of procollagen III is shown to be decreased in Ehlers-Danlos VIIC. Thus, the dogma that procollagen I and procollagen III N-proteinase activities are provided by separate enzymes appears to be false, whereas the phenotypes of dermatosparaxis and Ehlers-Danlos VIIC may arise from defects in both type I and type III collagen biosynthesis. PMID- 12646580 TI - A fusion protein of the gp130 and interleukin-6Ralpha ligand-binding domains acts as a potent interleukin-6 inhibitor. AB - Interleukin (IL)-6 is involved in the maintenance and progression of several diseases such as multiple myeloma, rheumatoid arthritis, or osteoporosis. The present work aims at the development of an IL-6 inhibitor for the use in anti cytokine therapies. The IL-6 receptor is composed of two different subunits, an alpha-subunit (IL-6Ralpha) that binds IL-6 with low affinity and a beta-subunit (gp130) that binds the IL-6.IL-6Ralpha complex with high affinity and as a result triggers intracellular signaling. In its soluble form, gp130 is a natural antagonist that neutralizes IL-6.soluble IL-6Ralpha complexes. It was our strategy to appropriately fuse the two receptor subunit fragments involved in IL 6 receptor complex formation to bind IL-6 with high affinity and to antagonize its effects. The ligand-binding domains of gp130 (D1-D2-D3) and IL-6Ralpha (D2 D3) were connected using three different linkers. The resulting constructs were expressed in stably transfected insect cells and tested for their ability to inhibit IL-6 activity in several in vitro systems. All fusion proteins were strong inhibitors of IL-6 signaling and abrogated IL-6-induced phosphorylation of STAT3, proliferation of transfected Ba/F3 cells, and induction of acute-phase protein synthesis. As intended, the fused receptors were much more effective than the separately expressed soluble receptor proteins. The fusion protein strategy presented here can also be applied to other cytokines that signal via receptors composed of two different subunits to design new potent inhibitors for anti cytokine therapies. PMID- 12646581 TI - Secretin activation of chromogranin A gene transcription. Identification of the signaling pathways in cis and in trans. AB - Secretin evokes catecholamine secretion from PC12 pheochromocytoma cells. We tested whether secretin activates transcription of the major vesicular core protein chromogranin A (CgA). Secretin stimulated both endogenous CgA gene transcription (approximately 4-6-fold) as well as transfected CgA promoter activity (approximately 8-10-fold; EC50, approximately 7 nm) in PC12 cells. Studies on CgA promoter 5'-deletion mutant/luciferase reporter constructs, point mutations of the CgA cAMP response element (CRE), and their transfer to a heterologous promoter implicated CRE in cis as both necessary and sufficient for secretin-stimulated CgA gene transcription. Secretin-induced CgA gene transcription was inhibited/abolished by cytosolic Ca2+ chelation, chemical blockade of phospholipase C, protein kinase A (PKA), or mitogen-activated protein (MAP) kinase extracellular signal regulated kinase (ERK) 1/2 and the expression of dominant negative mutants of ERK1/2, CRE binding protein (CREB) kinase RSK2, or CREB. Secretin also augmented (approximately 4-fold) phosphorylation of ERK1/2. Trans-activation (approximately 21-fold) of GAL4-CREB fusion protein by secretin indicates involvement of CREB in secretin signaling to gene transcription. Electrophoretic mobility shift assays also identified CREB as the mediator of secretin-induced CgA gene transcription, and pCREB supershifts indicated Ser-133 as the active CREB moiety in vitro. This conclusion was reinforced in vivo by results of chromatin pCREB immunoprecipitation assays. We conclude that secretin signals to CgA gene transcription through the CRE domain in cis and through cAMP, Ca2+, PKA, MAP kinase, and the transcription factor CREB in trans. Thus, multiple signal transduction pathways seem to subserve the function of stimulus-transcription coupling after this peptidergic stimulus to chromaffin cells. PMID- 12646582 TI - The direct interaction of phospholipase C-gamma 1 with phospholipase D2 is important for epidermal growth factor signaling. AB - The epidermal growth factor (EGF) receptor has an important role in cellular proliferation, and the enzymatic activity of phospholipase C (PLC)-gamma1 is regarded to be critical for EGF-induced mitogenesis. In this study, we report for the first time a phospholipase complex composed of PLC-gamma1 and phospholipase D2 (PLD2). PLC-gamma1 is co-immunoprecipitated with PLD2 in COS-7 cells. The results of in vitro binding analysis and co-immunoprecipitation analysis in COS-7 cells show that the Src homology (SH) 3 domain of PLC-gamma1 binds to the proline rich motif within the Phox homology (PX) domain of PLD2. The interaction between PLC-gamma1 and PLD2 is EGF stimulation-dependent and potentiates EGF-induced inositol 1,4,5-trisphosphate (IP(3)) formation and Ca(2+) increase. Mutating Pro 145 and Pro-148 within the PX domain of PLD2 to leucines disrupts the interaction between PLC-gamma1 and PLD2 and fails to potentiate EGF-induced IP(3) formation and Ca(2+) increase. However, neither PLD2 wild type nor PLD2 mutant affects the EGF-induced tyrosine phosphorylation of PLC-gamma1. These findings suggest that, upon EGF stimulation, PLC-gamma1 directly interacts with PLD2 and this interaction is important for PLC-gamma1 activity. PMID- 12646583 TI - Poly(A)-binding protein positively affects YB-1 mRNA translation through specific interaction with YB-1 mRNA. AB - The major protein of cytoplasmic mRNPs from rabbit reticulocytes, YB-1, is a member of an ancient family of proteins containing a common structural feature, cold-shock domain. In eukaryotes, this family is represented by multifunctional mRNA/Y-box DNA-binding proteins that control gene expression at different stages. To address possible post-transcriptional regulation of YB-1 gene expression, we examined effects of exogenous 5'- and 3'-untranslatable region-containing fragments of YB-1 mRNA on its translation and stability in a cell-free system. The addition of the 3' mRNA fragment as well as its subfragment I shut off protein synthesis at the initiation stage without affecting mRNA stability. UV cross-linking revealed four proteins (69, 50, 46, and 44 kDa) that specifically interacted with the 3' mRNA fragment; the inhibitory subfragment I bound two of them, 69- and 50-kDa proteins. We have identified these proteins as PABP (poly(A) binding protein) (69 kDa) and YB-1 (50 kDa) and demonstrated that titrating out of PABP by poly(A) strongly and specifically inhibits YB-1 mRNA cap(+)poly(A)(-) translation in a cell-free system. Thus, PABP is capable of positively affecting YB-1 mRNA translation in a poly(A) tail-independent manner. PMID- 12646584 TI - Stimulation of proliferation of rat hepatic stellate cells by galectin-1 and galectin-3 through different intracellular signaling pathways. AB - We found that the expression of galectin-1 and galectin-3 was significantly up regulated in hepatic stellate cells (HSCs) both in the course of their transdifferentiation into myofibroblasts, a process of "self-activation," and in the fibrosis of liver tissues. Recombinant galectin-1 and galectin-3 stimulated the proliferation of cultured HSCs via the MEK1/2-ERK1/2 signaling pathway. However, galectin-3 utilized protein kinases C and A to induce this process, whereas galectin-1 did not. We also found that thiodigalactoside, a potent inhibitor of beta-galactoside binding, attenuated the effects of both galectins. In addition, galectin-1, but not galectin-3, promoted the migration of HSCs. Thus, it appears that galectin-1 and galectin-3, generated by activated HSCs, could participate in beta-galactoside binding and induce different intracellular signaling pathways leading to the proliferation of HSCs. PMID- 12646585 TI - Control of cell polarity in fission yeast by association of Orb6p kinase with the highly conserved protein methyltransferase Skb1p. AB - In the fission yeast Schizosaccharomyces pombe, proper establishment and maintenance of cell polarity require Orb6p, a highly conserved serine/threonine kinase involved in regulating both cell morphogenesis and cell cycle control. Orb6p localizes to the cell tips during interphase and to the cell septum during mitosis. To investigate the mechanisms involved in Orb6p function, we conducted a two-hybrid screen to identify proteins that interact with Orb6p. Using this approach, we identified Skb1p, a highly conserved protein methyltransferase that has been implicated previously in cell cycle control, in the coordination of cell cycle progression with morphological changes, and in hyperosmotic stress response. We found that Skb1p associates with Orb6p in S. pombe cells and that the two proteins interact directly in vitro. Loss of Skb1p exacerbates the phenotype of orb6 mutants, suggesting that Skb1p and Orb6p functionally interact in S. pombe cells. Our results suggest that Skb1p affects the intracellular localization of Orb6p and that loss of Skb1p leads to a redistribution of the Orb6p kinase away from the cell tips. Furthermore, we found that Orb6p kinase activity is strongly increased following exposure to salt shock, suggesting that Orb6p has a role in cell response to hyperosmotic stress. Previous studies have shown that Skb1p interacts with the fission yeast p21-activated kinase homologue Pak1p/Shk1p to regulate cell polarity and cell cycle progression. Our findings identify Orb6p as an additional target for Skb1p and suggest a novel function for Skb1p in the control of cell polarity by regulating the subcellular localization of Orb6p. PMID- 12646586 TI - Expression of the telomeric repeat binding factor gene NgTRF1 is closely coordinated with the cell division program in tobacco BY-2 suspension culture cells. AB - Telomeres are vital for preserving chromosome integrity during cell division. Several genes encoding potential telomere-binding proteins have recently been identified in higher plants, but nothing is known about their function or regulation during cell division. In this study, we have isolated and characterized a cDNA clone, pNgTRF1, encoding a putative double-stranded telomeric repeat binding factor of Nicotiana glutinosa, a diploid tobacco plant. The predicted protein sequence of NgTRF1 (Mr = 75,000) contains a single Myb-like domain with significant homology to a corresponding motif in human TRF1/Pin2 and TRF2. Gel retardation assays revealed that bacterially expressed full-length NgTRF1 was able to form a specific complex only with probes containing three or more contiguous telomeric TTTAGGG repeats. The Myb-like domain of NgTRF1 is essential, but not sufficient, to bind the telomeric repeat sequence. The glutamine-rich extreme C-terminal region, which does not exist in animal proteins, was additionally required to form a specific telomere-protein complex. The dissociation constant (Kd) of the Myb motif plus the glutamine-rich domain of NgTRF1 to the two-telomeric repeat sequence was evaluated to be 4.5 +/- 0.2 x 10 9 m, which is comparable to that of the Myb domain of human TRF1. Expression analysis showed that NgTRF1 gene activity was inversely correlated with the cell division capacity of tobacco root cells and during the 9-day culture period of BY 2 suspension cells, while telomerase activity was positively correlated with cell division. In synchronized BY-2 cells, NgTRF1 was selectively expressed in G1 phase, whereas telomerase activity peaked in S phase. These findings suggest that telomerase activity and NgTRF1 expression are differentially regulated in an opposing fashion during growth and cell division in tobacco plants. The possible physiological functions of NgTRF1 in tobacco cells are also discussed. PMID- 12646587 TI - An NS3 serine protease inhibitor abrogates replication of subgenomic hepatitis C virus RNA. AB - The hepatitis C virus (HCV) NS3 protease is essential for polyprotein maturation and viral propagation, and it has been proposed as a suitable target for antiviral drug discovery. An N-terminal hexapeptide cleavage product of a dodecapeptide substrate identified as a weak competitive inhibitor of the NS3 protease activity was optimized to a potent and highly specific inhibitor of the enzyme. The effect of this potent NS3 protease inhibitor was evaluated on replication of subgenomic HCV RNA and compared with interferon-alpha (IFN-alpha), which is currently used in the treatment of HCV-infected patients. Treatment of replicon-containing cells with the NS3 protease inhibitor or IFN-alpha showed a dose-dependent decrease in subgenomic HCV RNA that reached undetectable levels following a 14-day treatment. Kinetic studies in the presence of either NS3 protease inhibitor or IFN-alpha also revealed similar profiles in HCV RNA decay with half-lives of 11 and 14 h, respectively. The finding that an antiviral specifically targeting the NS3 protease activity inhibits HCV RNA replication further validates the NS3 enzyme as a prime target for drug discovery and supports the development of NS3 protease inhibitors as a novel therapeutic approach for HCV infection. PMID- 12646588 TI - The Ski-binding protein C184M negatively regulates tumor growth factor-beta signaling by sequestering the Smad proteins in the cytoplasm. AB - Ski is a transcriptional co-repressor and is involved in the negative regulation of tumor growth factor-beta (TGF-beta) signaling. To understand more fully the role of Ski in TGF-beta signaling, we searched for novel Ski-interacting proteins. The identified C184M protein consists of 189 amino acids and contains the leucine-rich region. An association between Ski and C184M involving the leucine-rich region of C184M and the C-terminal coiled-coil motif of Ski was confirmed by glutathione S-transferase pull-down and immunoprecipitation assays. The C184M protein is located in the cytosol, and the C184M and Ski signals are co localized in the cytoplasm when C184M was co-expressed with Ski in CV-1 cells. The cytoplasmic C184M-Ski complex inhibited the nuclear translocation of Smad2. Consistent with this, the activity of promoter containing the Smad-binding sites was repressed by C184M, and the TGF-beta-induced growth inhibition of mink lung Mv1Lu cells was attenuated by the ectopic expression of C184M. Thus, C184M inhibits TGF-beta signaling in concert with Ski. In hepatocytes, which express significant levels of C184M, the Ski signals were found only in the cytoplasm, supporting the notion that C184M forms a complex with Ski in the cytosol. PMID- 12646589 TI - Negative interactions in close relationships: introduction to a special section. PMID- 12646590 TI - Negative interactions in close relationships across the life span. AB - This article examines age differences in positive, but especially negative, interactions in close relationships. Two community-based representative samples of people from 13 to 96 years of age from the United States and Japan were used to explore these relationships. Analyses indicate stability across age in positive interactions but a general decline in negative interactions in both countries. Three possible explanations for this age difference in negative interactions were examined: social maturity, familiarity, and contact frequency. Results provide most support for the contact frequency explanation. And finally, intercorrelations across relationships, that is, spouse, mother, father, child, and same-gender best friend, indicate moderate to high intercorrelations across all ages with a slight increase among older Japanese age groups. These results can be summarized as lending support to a generalized reduction in negative interactions with age but exceptions are noted in specific relationships and cultural traditions. PMID- 12646591 TI - Do age differences in close and problematic family ties reflect the pool of available relatives? AB - Older adults consistently report having fewer close and fewer problematic social ties than do younger adults. Yet, prior studies have not explicitly examined associations between the availability of living relatives and the nature of family ties in later life. One hundred sixty-four individuals ages 13 to 99 described their networks of close and problematic social ties. Then they provided information about specific living relatives (e.g., spouse, mother, father, sons, daughters, siblings, grandparents, and grandchildren). Older individuals reported more living relatives than did adolescents or young adults. Although individuals of all ages tended to name the majority of living relatives as close social contacts, most adults did not name all available relatives as close contacts. Age differences emerged with regard to naming relatives as problematic social ties. Older adults were less likely to indicate that their relatives caused problems. Findings are discussed in terms of an extension of investment theory, the availability hypothesis, which suggests that individuals with few living relatives are likely to view more of these relatives as close ties and fewer of these relatives as problematic. PMID- 12646592 TI - Negative interaction in late life: issues in the stability and generalizability of conflict across relationships. AB - This study used an inductive approach to probe more deeply into the nature of negative interaction in late life. The data come from a nationwide longitudinal survey of older adults (N = 515). We began with two empirical observations and derived theoretical propositions from them that have not been explored extensively in mainstream gerontological research on negative interaction. First, the findings reveal that unpleasant interaction is quite stable over a 6-year period. This suggests that interpersonal difficulties are chronic stressors that may arise because at least some older people are involved in conflicted relationships that are difficult to terminate. The second empirical finding reveals that older adults who encounter interpersonal problems in one social relationship (e.g., with children) tend to encounter them in others as well (e.g., with friends). This suggests that some older people may play a role in creating the negative interactions they encounter. PMID- 12646593 TI - Exposure and reactivity to negative social exchanges: a preliminary investigation using daily diary data. AB - Negative social exchanges have the potential to detract from well-being in later life, but the factors that influence older adults' vulnerability to such exchanges remain poorly understood. Two dimensions of vulnerability, exposure and reactivity, were investigated using daily diary data collected at two time points from a sample of community-residing older adults (N = 129). Three categories of predictors were examined: individual characteristics, life stress, and social network characteristics. Greater exposure to negative social exchanges was related to greater life stress and to less supportive networks. Greater reactivity was related to lower self-esteem, less supportive networks, less satisfying friendships and family relationships, and surprisingly, to less life stress. Longitudinal changes in exposure and reactivity were related to changes in friendship satisfaction, the positive functions performed by the network, and health status. The findings suggest that personal characteristics and life circumstances play a role in influencing older adults' vulnerability to negative social exchanges. PMID- 12646594 TI - Associations of stressors and uplifts of caregiving with caregiver burden and depressive mood: a meta-analysis. AB - In the present meta-analysis, we integrated findings from 228 studies on the association of six caregiving-related stressors and caregiving uplifts with burden and depressed mood. Care recipients' behavior problems showed stronger associations with caregiver outcomes than other stressors did. The size of the relationships varied by sample characteristics: Amount of care provided and care receivers' physical impairments were less strongly related to burden and depression for dementia caregivers than for caregivers of nondemented older adults. For spouse caregivers, physical impairments and care recipients' behavior problems had a stronger relationship to burden than for adult children. Furthermore, we found evidence that the association of caregiver burden with the number of caregiving tasks, perceived uplifts of caregiving, and the level of physical impairment of the care receiver were stronger in probability samples than in convenience samples. PMID- 12646595 TI - Predictors of agitation in nursing home residents. AB - Agitation in nursing home residents presents a serious challenge to caregivers and may place residents at risk for harm. Understanding the etiology of agitation can assist clinicians in developing nonpharmacologic interventions for preventing and treating this problem. The purpose of this study was to examine independent and common predictors of resident agitation with structural equation modeling. Agitation was measured with both a standardized staff report rating scale and direct behavioral observation. No indirect or mediating effects were found. Cognitive impairment, vision and hearing impairment, and gender were found to be independent predictors of agitation as measured by direct behavioral observation. Only cognitive impairment was found to be predictive of agitation as measured by the standardized staff report scale. An unexpected finding was that vision impairment appeared to exert a protective effect for agitation in these severely cognitively impaired residents. The clinical implications of these findings are discussed as well as the relative merits of the two methods of measuring agitation. PMID- 12646596 TI - The relation between everyday activities and successful aging: a 6-year longitudinal study. AB - OBJECTIVES: Activity has long been thought to be related to successful aging. This study was designed to examine longitudinally the relation between everyday activities and indicators of successful aging, namely well-being, function, and mortality. METHODS: The study was based on the Aging in Manitoba Study, with activity being measured in 1990 and function, well-being, and mortality assessed in 1996. Well-being was measured in terms of life satisfaction and happiness; function was defined in terms of a composite measure combining physical and cognitive function. RESULTS: Regression analyses indicated that greater overall activity level was related to greater happiness, better function, and reduced mortality. Different activities were related to different outcome measures; but generally, social and productive activities were positively related to happiness, function, and mortality, whereas more solitary activities (e.g., hand-work hobbies) were related only to happiness. DISCUSSION: These findings highlight the importance of activity in successful aging. The results also suggest that different types of activities may have different benefits. Whereas social and productive activities may afford physical benefits, as reflected in better function and greater longevity, more solitary activities, such as reading, may have more psychological benefits by providing a sense of engagement with life. PMID- 12646597 TI - English language skills, ethnic concentration, and household composition: older Mexican immigrants. AB - OBJECTIVES: The authors examine the living arrangements of older Mexican immigrants to demonstrate how the formation of complex households is related to English language proficiency. Specifically, they examine whether the ability to use the English language is a determinant of an older Mexican immigrant persons' capacity to maintain residential independence. They also examine how living in communities with a relatively high proportion of Hispanic persons impacts household structure. METHODS: Data from the 1990 U.S. Census of Population are used to evaluate multilevel multinomial logistic regression models of living arrangements among older Mexican immigrants. RESULTS: The authors found that strong English language skills increase the likelihood of living independently and increase the likelihood of being the head of a household. They also found that living in an area with a relatively high proportion of Hispanic persons increases the likelihood that older Mexican immigrants will live independently as compared with living with others without headship. Finally, interaction models were examined, demonstrating that living in a community with a relatively high proportion of Hispanics reduces the strength of the relationship between English language proficiency and living arrangement outcomes for older Mexican immigrants. DISCUSSION: The authors discuss the implications of their findings and point out possible avenues for further research. PMID- 12646598 TI - Social networks, social integration, and social engagement determine cognitive decline in community-dwelling Spanish older adults. AB - OBJECTIVES: To examine the influence of social networks and social engagement on cognitive decline in a population-based cohort of elderly people, and to assess gender differences in the effect of social relations on cognition. METHODS: A longitudinal study of community-dwelling people over 65 was carried out. Cognitive function (orientation and memory) in 1997 and cognitive decline (absent, mild, and severe) over 4 years (1993-1997) were assessed using an instrument previously validated for populations with a low level of education. The effect of social networks, social integration, and social engagement with friends, children, and relatives on cognitive function and cognitive decline was estimated by multiple linear and logistic regressions after adjusting for age, sex, education, depressive symptoms, systolic and diastolic blood pressure, and functional status. RESULTS: Poor social connections, infrequent participation in social activities, and social disengagement predict the risk of cognitive decline in elderly individuals. The probability of cognitive decline was lower for both men and women with a high frequency of visual contact with relatives and community social integration. Engagement with friends seemed to be protective for cognitive decline in women but not in men. DISCUSSION: This longitudinal study indicates that few social ties, poor integration, and social disengagement are risk factors for cognitive decline among community-dwelling elderly persons. The nature of the ties that influence cognition may vary in men and women. PMID- 12646599 TI - Socioeconomic status and age identity: the role of dimensions of health in the subjective construction of age. AB - OBJECTIVES: This study examines health inequality as a potential explanation for socioeconomic differences in age identity. The following dimensions of health are examined: comparative self-rated health, self-assessed changes in physiological well-being, prospective self-rated health, perceived control over health, chronic conditions, and parents' health. Components of health also are explored as possible mediators of age differences in the effect of socioeconomic status on age identity. METHODS: Using the National Survey of Midlife Development in the United States, ordinary least squares regression models of age identity are estimated for a representative sample of the noninstitutionalized U.S. population between the ages of 25 and 74 (n = 2,864). RESULTS: The older identities held by the less socioeconomically advantaged are explained by their worse health, particularly their less favorable predictions of future health, compared with their wealthier peers. Differences in age identity by education and perceived financial well-being are greatest among older adults; however, health only partially accounts for these age patterns. DISCUSSION: This study reveals that health inequalities shape the subjective experience of aging. Further work using longitudinal data is needed to determine the precise causal pathways linking socioeconomic status, health, and age identity. PMID- 12646601 TI - An examination of the impact of health on wealth depletion in elderly individuals. AB - OBJECTIVES: This study investigates the effects of new health events and existing health conditions on wealth depletion in elderly individuals. METHODS: A model deriving from life-cycle theory is proposed and estimated using Waves 1 and 2 of the Asset and Health Dynamics of the Oldest Old (AHEAD) data set. RESULTS: Both new health events and existing health conditions significantly influence wealth depletion of elders, but their impacts differ across marital status. Whereas an occurrence of new health events brought wealth depletion of elders in married households, having existing chronic health conditions was associated with wealth depletion of elders in single households. DISCUSSION: Poor health, both a new health event and existing chronic conditions, leads to considerable wealth depletion in elderly individuals. Considering the significant impacts of health on wealth, the public needs to be better informed of potential health events in later life and the associated financial burden. Additional health insurance plays an important role in preventing elders from financial hardship. PMID- 12646600 TI - The life course of severe obesity: does childhood overweight matter? AB - OBJECTIVES: A life course perspective is used to examine the epidemiology of severe obesity in adulthood, defined as a body mass index > or = 35 kg/m(2). METHODS: Data from adults in the National Health and Nutrition Examination Survey I: Epidemiologic Followup Study, including their reports of childhood overweight, were used to examine the risk of severe obesity and mortality over 20 years (N = 6,767). All multivariate models control for age, sex, race, smoking, and socioeconomic resources. RESULTS: Childhood overweight was significantly associated with severe obesity for both women and men, although the effect was stronger for men. The prevalence of severe obesity was highest between 45 and 64 years of age than for persons under 45 or over 65, and higher for African Americans than White Americans. Childhood overweight was associated with lower mortality risk for women, but not for men. Mortality risk was higher for persons with severe obesity (relative risk = 1.571, 95% confidence interval = 1.335 1.849, p <.001). DISCUSSION: The findings demonstrate the importance of childhood overweight as a risk factor for severe obesity over the life course. Nevertheless, overweight children who did not become severely obese were not at greater risk of mortality. PMID- 12646602 TI - Analysis of the reliability of the modified caregiver strain index. AB - OBJECTIVES: The Caregiver Strain Index (CSI) is a 13-item index originally developed in the early 1980s to screen for caregiver strain after hospital discharge of an elderly family member. This study examined the reliability of a modified CSI for applications with contemporary long-term family caregivers. METHODS: As part of a larger family caregiver study using focus groups, telephone interviews, and mailed surveys, test-retest procedures were included to examine the reliability of the Modified CSI. The sample consisted of 158 family caregivers, with an average age of 61 years. Targeted recruitment resulted in participants from diverse racial/ethnic backgrounds. Minority caregivers represented 25% of the sample. RESULTS: The Modified CSI has slightly better internal reliability (alpha =.90) than the original Index, as reported in 1983 (alpha =.86). The two-week test-retest reliability is.88. No prior test-retest data were available for comparison. DISCUSSION: The Modified CSI can be a useful method for detecting strain levels among informal caregivers, and is easily administered and scored. The CSI continues to be a useful measure of caregiver strain for long-term care research and practice. PMID- 12646604 TI - Cutting edge: a Toll-like receptor 2 polymorphism that is associated with lepromatous leprosy is unable to mediate mycobacterial signaling. AB - Toll-like receptors (TLRs) are key mediators of the innate immune response to microbial pathogens. We investigated the role of TLRs in the recognition of Mycobacterium leprae and the significance of TLR2Arg(677)Trp, a recently discovered human polymorphism that is associated with lepromatous leprosy. In mice, TNF-alpha production in response to M. leprae was essentially absent in TLR2-deficient macrophages. Similarly, human TLR2 mediated M. leprae-dependent activation of NF-kappaB in transfected Chinese hamster ovary and human embryonic kidney 293 cells, with enhancement of this signaling in the presence of CD14. In contrast, activation of NF-kappaB by human TLR2Arg(677)Trp was abolished in response to M. leprae and Mycobacterium tuberculosis. The impaired function of this TLR2 variant provides a molecular mechanism for the poor cellular immune response associated with lepromatous leprosy and may have important implications for understanding the pathogenesis of other mycobacterial infections. PMID- 12646605 TI - Cutting edge: CD40-induced expression of recombination activating gene (RAG) 1 and RAG2: a mechanism for the generation of autoaggressive T cells in the periphery. AB - It has been speculated that autoimmune diseases are caused by failure of central tolerance. However, this remains controversial. We have suggested that CD40 expression identifies autoaggressive T cells in the periphery of autoimmune prone mice. In this study, we report that CD40 was cloned from autoaggressive T cells and that engagement induces expression and nuclear translocation of the recombinases, recombination activating gene (RAG) 1 and RAG2 in the autoaggressive, but not in the nonautoaggressive, peripheral T cell population. Furthermore, we demonstrate that CD40 engagement induces altered TCR Valpha, but not Vbeta, expression in these cells. Therefore, CD40-regulated expression of RAG1 and RAG2 in peripheral T cells may constitute a novel pathway for the generation of autoaggressive T cells. PMID- 12646606 TI - IL-9 enhances the growth of human mast cell progenitors under stimulation with stem cell factor. AB - We examined the effects of IL-9 on human mast cell development from CD34(+) cord blood (CB) and peripheral blood cells in serum-deprived cultures. IL-9 apparently enhanced cell production under stimulation with stem cell factor (SCF) from CD34(+) CB cells. A great majority of the cultured cells grown with SCF + IL-9 became positive for tryptase at 4 wk. In methylcellulose cultures of CD34(+) CB cells, IL-9 increased both the number and size of mast cell colonies grown with SCF. Furthermore, SCF + IL-9 caused an exclusive expansion of mast cell colony forming cells in a 2-wk liquid culture of CD34(+) CB cells, at a level markedly greater than for SCF alone. Clonal cell cultures and RT-PCR analysis showed that the targets of SCF + IL-9 were the CD34(+)CD38(+) CB cells rather than the CD34(+)CD38(-) CB cells. IL-9 neither augmented the SCF-dependent generation of progeny nor supported the survival of 6-wk-cultured mast cells. Moreover, there was no difference in the appearance of tryptase(+) cells and histamine content in the cultured cells between SCF and SCF + IL-9. The addition of IL-9 increased numbers of mast cell colonies grown with SCF from CD34(+) peripheral blood cells in children with or without asthma. It is of interest that mast cell progenitors of asthmatic patients responded to SCF + IL-9 to a greater extent than those of normal controls. Taken together, IL-9 appears to act as a potent enhancer for the SCF-dependent growth of mast cell progenitors in humans, particularly asthmatic patients. PMID- 12646607 TI - CpG-A oligonucleotides induce a monocyte-derived dendritic cell-like phenotype that preferentially activates CD8 T cells. AB - Human B cells and plasmacytoid dendritic cells recognize CpG motifs within microbial DNA via Toll-like receptor 9. Two functionally distinct types of CpG motif containing oligonucleotides (CpG ODN) have been described, CpG-A and CpG-B. In contrast to CpG-B, CpG-A induces high amounts of type I IFN (IFN-alpha and IFN beta) in plasmacytoid dendritic cells. In the present study, we examined the effects of CpG-A on human primary monocytes. In PBMC stimulated with CpG-A and GM CSF, monocytes showed excellent survival, increased in size and granularity, and within 3 days developed a dendritic cell-like phenotype that was characterized by down-regulation of CD14, partial up-regulation of CCR7, and an increased surface expression of costimulatory and Ag-presenting molecules. This effect could be inhibited by a combination of blocking Abs to type I IFN, and no such CpG-A induced changes were observed in purified monocytes. Although IL-12 production by this dendritic cell-like phenotype required additional stimulation with CD40 ligand, this cell type spontaneously up-regulated IL-15 expression. Consistent with the known effect of IL-15 on effector and memory CD8 T cells, the frequency of CCR7(-)/CD45RA(-) CD8 T cells was selectively increased in allogeneic T cell assays. Furthermore, this dendritic cell type was more potent to support both the generation and the IFN-gamma production of autologous influenza matrix peptide specific memory CD8 T cells as compared with dendritic cells generated in the presence of GM-CSF and IL-4. In conclusion, monocytes exposed to the cytokine milieu provided by CpG-A rapidly develop a dendritic cell-like phenotype that is well equipped to support CD8 T cell responses. PMID- 12646608 TI - Expression of a constitutively active Stat6 in vivo alters lymphocyte homeostasis with distinct effects in T and B cells. AB - IL-4 is a critical cytokine in the regulation of immune responses and genesis of atopy. Engagement of the IL-4R activates multiple signaling pathways, including the transcription factor Stat6. Stat6-deficient mice demonstrate the importance of this factor in lymphocyte proliferation, gene expression, and Th cell differentiation. Recently, a mutant Stat6 (Stat6VT) was generated that is transcriptionally active independent of IL-4 stimulation. To determine the ability of a constitutively active Stat6 to mimic IL-4-stimulated responses, we have generated transgenic mice expressing Stat6VT under control of the CD2 locus control region, restricting expression to lymphoid populations. The phenotype of Stat6VT transgenic mice is similar, but not identical, to IL-4 transgenic mice, suggesting a critical role for Stat6-independent signaling pathways in the generation of some IL-4 responses in vivo. The expression of a constitutively active Stat6 in vivo increases surface expression of IL-4-induced genes and increases serum levels of IgG1 and IgE, compared with nontransgenic mice. Stat6VT expression increases Th2 differentiation in vivo and in vitro. Stat6VT expression also dramatically alters homeostasis of peripheral lymphocyte populations resulting in decreased CD3(+) cells and increased B220(+) cells, compared with nontransgenic littermates. Altered T and B cell populations correlate with an activated phenotype and increased cell death in transgenic T cell, but not B cell, populations. Together these results suggest that expression of a constitutively active Stat6 has distinct effects on B and T lymphocytes. PMID- 12646609 TI - Gangliosides from human melanoma tumors impair dendritic cell differentiation from monocytes and induce their apoptosis. AB - Gangliosides are ubiquitous membrane-associated glycosphingolipids, which are involved in cell growth and differentiation. Most tumor cells synthesize and shed large amounts of gangliosides into their microenvironment, and many studies have unraveled their immunosuppressive properties. In the present study we analyzed the effects of GM3 and GD3 gangliosides, purified from human melanoma tumors, on the differentiation of monocyte-derived dendritic cells (MoDC). At concentrations close to those detected in the sera from melanoma patients, both gangliosides dose-dependently inhibit the phenotypic and functional differentiation of MoDC, as assessed by a strong down-regulation of CD1a, CD54, CD80, and CD40 Ags and impaired allostimulatory function on day 6 of culture. Furthermore, GM3 and GD3 gangliosides decreased the viable cell yield and induced significant DC apoptosis. Finally, addition of GD3 to differentiating DC impaired their subsequent maturation induced by CD154. The resulting DC produced low amounts of IL-12 and large amounts of IL-10, a cytokine pattern that might hamper an efficient antitumor immune response. In conclusion, the results demonstrate that gangliosides impair the phenotypic and functional differentiation of MoDC and induce their apoptosis, which may be an additional mechanism of human melanoma escape. PMID- 12646610 TI - IL-4 potentiates activated T cell apoptosis via an IL-2-dependent mechanism. AB - Activation-induced cell death (AICD) of T cells is one of the major mechanisms of peripheral tolerance. The regulation of AICD by IL-4 is poorly understood. In this study, we report that AICD in IL-4-deficient T cells is significantly reduced compared with that in wild-type T cells. This impaired AICD correlates with the failure to induce degradation of cellular FLIP. IL-4-mediated enhancement of AICD and cellular FLIP degradation requires a Janus kinase/STAT-6 signaling pathway. Unexpectedly, these effects of IL-4 could be blocked by a neutralizing anti-IL-2 Ab, and addition of rIL-2 could completely restore the defective AICD in IL-4-deficient T cells. Furthermore, IL-4 regulates the T cell thresholds for IL-2 signaling during AICD. These data suggest that IL-4 promotes AICD via an IL-2-dependent mechanism. PMID- 12646611 TI - Presentation by recycling MHC class II molecules of an influenza hemagglutinin derived epitope that is revealed in the early endosome by acidification. AB - We investigated the roles of nascent and recycling MHC class II molecules (MHC II) in the presentation of two well-defined I-E(d)-restricted epitopes that are within distinct regions of the influenza virus hemagglutinin (HA) protein. The site 3 epitope (S3; residues 302-313) lies in the stalk region that unfolds in response to mild acidification, while the site 1 epitope (S1; residues 107-119) is situated in the stable globular domain. In a murine B lymphoma cell line and an I-E(d)-transfected fibroblast cell line, presentation from inactivated virus of S3 is inhibited by primaquine, a compound that prevents recycling of cell surface proteins, including MHC II, while S1 presentation is unaffected. In contrast, brefeldin A, an agent that inhibits exit of proteins from the endoplasmic reticulum, selectively inhibited S1 presentation without affecting S3 presentation, suggesting that S1 presentation requires nascent MHC II. The use of agents that perturb endosomal function revealed a requirement for acidification of internalized viral particles for presentation of both epitopes. Notably, all compounds tested had similar effects on presentation of the two epitopes derived from endogenously synthesized HA. Thus, recycling I-E(d) molecules appear to be crucial for capturing and presenting an epitope that is revealed in mild acidic conditions following the uptake of virions or the synthesis of Ag, while nascent I-E(d) molecules are required for presentation of a second epitope located in a structurally constrained region of the same polypeptide. Viral glycoproteins, such as HA, may have been a major impetus for the evolutionary establishment of this recycling pathway. PMID- 12646612 TI - Two developmentally distinct populations of dendritic cells inhabit the adult mouse thymus: demonstration by differential importation of hematogenous precursors under steady state conditions. AB - Although a variety of lymphoid and myeloid precursors can generate thymic dendritic cells (DCs) under defined experimental conditions, the developmental origin(s) of DCs in the steady state thymus is unknown. Having previously used selective combinations of normal, parabiotic, and radioablated mice to demonstrate that blood-borne prothymocytes are imported in a gated and competitive manner, we used a similar approach in this study to investigate the importation of the hematogenous precursors of thymic DCs. The results indicate that two developmentally distinct populations of DC precursors normally enter the adult mouse thymus. The first population is indistinguishable from prothymocytes according to the following criteria: 1) inefficient (<20%) exchange between parabiotic partners; 2) gated importation by the thymus; 3) competitive antagonism for intrathymic niches; 4) temporally linked generation of thymocytes and CD8alpha(high) DCs; and 5) absence from prothymocyte-poor blood samples. The second population differs diametrically from prothymocytes in each of these properties, and appears to enter the thymus in at least a partially differentiated state. The resulting population of DCs has a CD8alpha(-/low) phenotype, and constitutes approximately 50% of total thymic DCs. The presence of two discrete populations of DCs in the steady state thymus implies functional heterogeneity consistent with evidence implicating lymphoid DCs in the negative selection of effector thymocytes and myeloid DCs in the positive selection of regulatory thymocytes. PMID- 12646613 TI - Prevention of autoantibody-mediated Graves'-like hyperthyroidism in mice with IL 4, a Th2 cytokine. AB - Graves' hyperthyroidism has long been considered to be a Th2-type autoimmune disease because it is directly mediated by autoantibodies against the thyrotropin receptor (TSHR). However, several lines of evidence have recently challenged this concept. The present study evaluated the Th1/Th2 paradigm in Graves' disease using a recently established murine model involving injection of adenovirus expressing the TSHR (AdCMVTSHR). Coinjection with adenovirus expressing IL-4 (AdRGDCMVIL-4) decreased the ratio of Th1/Th2-type anti-TSHR Ab subclasses (IgG2a/IgG1) and suppressed the production of IFN-gamma by splenocytes in response to TSHR Ag. Importantly, immune deviation toward Th2 was accompanied by significant inhibition of thyroid-stimulating Ab production and reduction in hyperthyroidism. However, in a therapeutic setting, injection of AdRGDCMVIL-4 alone or in combination with AdCMVTSHR into hyperthyroid mice had no beneficial effect. In contrast, coinjection of adenoviruses expressing IL-12 and the TSHR promoted the differentiation of Th1-type anti-TSHR immune responses as demonstrated by augmented Ag-specific IFN-gamma secretion from splenocytes without changing disease incidence. Coinjection of adenoviral vectors expressing IL-4 or IL-12 had no effect on the titers of anti-TSHR Abs determined by ELISA or thyroid-stimulating hormone-binding inhibiting Ig assays, suggesting that Ab quality, not quantity, is responsible for disease induction. Our observations demonstrate the critical role of Th1 immune responses in a murine model of Graves' hyperthyroidism. These data may raise a cautionary note for therapeutic strategies aimed at reversing Th2-mediated autoimmune responses in Graves' disease in humans. PMID- 12646614 TI - The MHC class I-like IgG receptor controls perinatal IgG transport, IgG homeostasis, and fate of IgG-Fc-coupled drugs. AB - Abs of the IgG isotype are efficiently transported from mother to neonate and have an extended serum t(1/2) compared with Abs of other isotypes. Circumstantial evidence suggests that the MHC class I-related protein, the neonatal FcR (FcRn), is the FcR responsible for both in vivo functions. To understand the phenotypes imposed by FcRn, we produced and analyzed mice with a defective FcRn gene. The results provide direct evidence that perinatal IgG transport and protection of IgG from catabolism are mediated by FcRn, and that the latter function is key to IgG homeostasis, essential for generating a potent IgG response to foreign Ags, and the basis of enhanced efficacy of Fc-IgG-based therapeutics. FcRn is therefore a promising therapeutic target for enhancing protective humoral immunity, treating autoimmune disease, and improving drug efficacy. PMID- 12646616 TI - The mechanism of CD47-dependent killing of T cells: heterotrimeric Gi-dependent inhibition of protein kinase A. AB - CD47 has been implicated in both positive and negative regulation of T cells as well as in T cell death. To clarify the role of CD47 in T cell function, we have studied the mechanism of T cell death in response to CD47 ligands, including mAb 1F7, thrombospondin-1, and a CD47 agonist peptide derived from it. CD47(-/-) Jurkat T cells (JINB8) were resistant to killing by all three ligands, indicating the essential role of CD47. Primary human T cells were also killed by CD47 ligands, but only after activation with anti-CD3. CD47-mediated cell death occurred without active caspases, DNA fragmentation, or Bcl-2 degradation. Pretreatment of Jurkat and primary T cells with pertussis toxin (PTX) prevented CD47-mediated death, indicating the involvement of G((i)alpha). Pretreatment of T cells with 8-bromo cAMP, forskolin, or 3-isobutyl-1-methylxanthine prevented the CD47-mediated apoptosis, and 1F7 dramatically reduced intracellular cAMP levels, an effect reversed with PTX. H89 and protein kinase A (PKA) inhibitor peptide, a specific PKA inhibitor, prevented rescue of T cells by PTX, 8-bromo cAMP, and forskolin, indicating a direct role for one or more PKA substrates. Thus, CD47 mediated killing of activated T cells occurs by a novel pathway involving regulation of cAMP levels by heterotrimeric G((i)alpha) with subsequent effects mediated by PKA. PMID- 12646615 TI - The B cell coreceptor CD22 associates with AP50, a clathrin-coated pit adapter protein, via tyrosine-dependent interaction. AB - The B cell coreceptor CD22 plays an important role in regulating signal transduction via the B cell Ag receptor. Studies have shown that surface expression of CD22 can be modulated in response to binding of ligand (i.e., mAb). Thus, it is possible that alterations in the level of CD22 expression following binding of natural ligand(s) may affect its ability to modulate the Ag receptor signaling threshold at specific points during B cell development and differentiation. Therefore, it is important to delineate the physiologic mechanism by which CD22 expression is controlled. In the current study, yeast two hybrid analysis was used to demonstrate that CD22 interacts with AP50, the medium chain subunit of the AP-2 complex, via tyrosine-based internalization motifs in its cytoplasmic domain. This interaction was further characterized using yeast two-hybrid analysis revealing that Tyr(843) and surrounding amino acids in the cytoplasmic tail of CD22 comprise the primary binding site for AP50. Subsequent studies using transfectant Jurkat cell lines expressing wild-type or mutant forms of CD22 demonstrated that either Tyr(843) or Tyr(863) is sufficient for mAb mediated internalization of CD22 and that these motifs are involved in its interaction with the AP-2 complex, as determined by coprecipitation of alpha adaptin. Finally, experiments were performed demonstrating that treatment of B cells with either intact anti-Ig Ab or F(ab')(2) blocks ligand-mediated internalization of CD22. In conclusion, these studies demonstrate that internalization of CD22 is dependent on its association with the AP-2 complex via tyrosine-based internalization motifs. PMID- 12646617 TI - Murine Flt3 ligand expands distinct dendritic cells with both tolerogenic and immunogenic properties. AB - Human Flt3 ligand can expand dendritic cells (DC) and enhance immunogenicity in mice. However, little is known about the effects of murine Flt3 ligand (mFlt3L) on mouse DC development and function. We constructed a vector to transiently overexpress mFlt3L in mice. After a single treatment, up to 44% of splenocytes became CD11c(+) and the total number of DC increased 100-fold. DC expansion effects lasted for >35 days. mFlt3L DC were both phenotypically and functionally distinct. They had increased expression of MHC and costimulatory molecules and expressed elevated levels of B220 and DEC205 but had minimal CD4 staining. mFlt3L DC also had a markedly altered cytokine profile, including lowered secretion of IL-6, IL-10, IFN-gamma, and TNF-alpha, but had a slightly increased capacity to stimulate T cells in vitro. However, in a variety of in vivo models, DC expanded by mFlt3L induced tolerogenic effects on T cells. Adoptive transfer of Ag-pulsed mFlt3L splenic DC to naive mice actually caused faster rates of tumor growth and induced minimal CTL compared with control DC. mFlt3L also failed to protect against tumors in which human Flt3 ligand was protective, but depletion of CD4(+) T cells restored tumor protection. Our findings 1) demonstrate that mFlt3L has distinct effects on DC development, 2) suggest an important role for mFlt3L in generating DC that have tolerogenic effects on T cells, and 3) may have application in immunotherapy in generating massive numbers of DC for an extended duration. PMID- 12646619 TI - Activated, but not resting, T cells can be recognized and killed by syngeneic NK cells. AB - We demonstrate that IL-2-activated NK cells or lymphokine-activated killer cells recognize and kill syngeneic CD4(+) and CD8(+) T cells that have been activated by APCs. Induction with APC required TCR-specific Ag, and lysis was perforin mediated. Brefeldin A, which disrupts protein transport, inhibited the sensitivity induced by activation. In BALB/c, expression of NKG2D ligands correlated with lysis and could be inhibited by brefeldin A. As well, addition of anti-NKG2D mAb to a killing assay completely abrogated lysis. Transduction of mouse NKG2D into a human NK cell line, YTSeco, conferred upon it the ability to kill activated BALB/c T cells, indicating that NKG2D is necessary for recognition. Our data provide a basis for studying a role for NK cells in T cell regulation. PMID- 12646618 TI - Toll-like receptor 3 mediates a more potent antiviral response than Toll-like receptor 4. AB - We have recently described an IFN regulatory factor 3-mediated antiviral gene program that is induced by both Toll-like receptor (TLR)3 and TLR4 ligands. In our current study, we show that activation of IFN/viral response gene expression in primary macrophage cells is stronger and prolonged with TLR3 stimulation compared with that of TLR4. Our data also reveal that the cytoplasmic tails of both TLR3 and TLR4 can directly interact with myeloid differentiation factor 88 (MyD88). However, although Toll/IL-1 receptor homology domain-containing adaptor protein/MyD88 adaptor-like is able to associate with TLR4, we were unable to detect any interaction between Toll/IL-1 receptor homology domain-containing adaptor protein/MyD88 adaptor-like and TLR3. By using quantitative real-time PCR assays, we found that TLR3 expression is inducible by both TLR3 and TLR4 ligands, while TLR4 expression is not inducible by these same stimuli. Furthermore, using cells derived from mice deficient in the IFN-alphabetaR, we show that both TLR3 and TLR4 require IFN-beta autocrine/paracrine feedback to induce TLR3 expression and activate/enhance genes required for antiviral activity. More specifically, a subset of antiviral genes is initially induced independent of IFN-beta, yet the cytokine further enhances expression at later time points. This was in contrast to a second set of genes (including TLR3) that is induced only after IFN-beta production. Taken together, our data argue that, despite both TLR3 and TLR4 being able to use IFN-beta to activate/enhance antiviral gene expression, TLR3 uses multiple mechanisms to enhance and sustain the antiviral response more strongly than TLR4. PMID- 12646620 TI - Ligation of retinoic acid receptor alpha regulates negative selection of thymocytes by inhibiting both DNA binding of nur77 and synthesis of bim. AB - Negative selection refers to the selective deletion of autoreactive thymocytes. Its molecular mechanisms have not been well defined. Previous studies in our laboratory have demonstrated that retinoic acids, physiological ligands for the nuclear retinoid receptors, selectively inhibit TCR-mediated death under in vitro conditions, and the inhibition is mediated via the retinoic acid receptor (RAR) alpha. The present studies were undertaken to investigate whether ligation of RARalpha leads to inhibition of TCR-mediated death in vivo and to identify the molecular mechanisms involved. Three models of TCR-mediated death were studied: anti-CD3-mediated death of thymocytes in wild-type mice, and Ag- and bacterial superantigen-driven thymocyte death in TCR-transgenic mice expressing a receptor specific for a fragment of pigeon cytochrome c in the context of the E(k) (class II MHC) molecule. Our data demonstrate that the molecular program of both anti CD3- and Ag-driven, but not that of superantigen-mediated apoptosis involves up regulation of nur77, an orphan nuclear receptor, and bim, a BH3-only member of the proapoptotic bcl-2 protein family, proteins previously implicated to participate in the negative selection. Ligation of RARalpha by the synthetic agonist CD336 inhibited apoptosis, DNA binding of nur77, and synthesis of bim induced by anti-CD3 or the specific Ag, but had no effect on the superantigen driven cell death. Our data imply that retinoids are able to inhibit negative selection in vivo as well, and they interfere with multiple steps of the T cell selection signal pathway. PMID- 12646621 TI - IL-10 mediates sigma 1 receptor-dependent suppression of antitumor immunity. AB - Sigma receptors are unique endoplasmic reticulum proteins that mediate signaling for a variety of drugs. We determined the effect of sigma(1) receptor agonists on immune responses in a syngeneic lung cancer model. Sigma(1) receptor agonists, including cocaine, up-regulated splenocyte IL-10 mRNA and protein production in vitro in a sigma receptor-dependent, pertussis toxin-sensitive manner. In vivo, sigma(1) receptor agonists promoted tumor growth and induced IL-10 at the tumor site. Increased tumor growth was prevented by administration of specific Abs to IL-10 or by administration of specific sigma(1) receptor antagonists. We report that sigma(1) receptor ligands, including cocaine, augment tumor growth through an IL-10 dependent mechanism. PMID- 12646622 TI - Infections that induce autoimmune diabetes in BBDR rats modulate CD4+CD25+ T cell populations. AB - Viruses are believed to contribute to the pathogenesis of autoimmune type 1A diabetes in humans. This pathogenic process can be modeled in the BBDR rat, which develops pancreatic insulitis and type 1A-like diabetes after infection with Kilham's rat virus (RV). The mechanism is unknown, but does not involve infection of the pancreatic islets. We first documented that RV infection of BBDR rats induces diabetes, whereas infection with its close homologue H-1 does not. Both viruses induced similar humoral and cellular immune responses in the host, but only RV also caused a decrease in splenic CD4(+)CD25(+) T cells in both BBDR rats and normal WF rats. Surprisingly, RV infection increased CD4(+)CD25(+) T cells in pancreatic lymph nodes of BBDR but not WF rats. This increase appeared to be due to the accumulation of nonproliferating CD4(+)CD25(+) T cells. The results imply that the reduction in splenic CD4(+)CD25(+) cells observed in RV-infected animals is virus specific, whereas the increase in pancreatic lymph node CD4(+)CD25(+) cells is both virus and rat strain specific. The data suggest that RV but not H-1 infection alters T cell regulation in BBDR rats and permits the expression of autoimmune diabetes. More generally, the results suggest a mechanism that could link an underlying genetic predisposition to environmental perturbation and transform a "regulated predisposition" into autoimmune diabetes, namely, failure to maintain regulatory CD4(+)CD25(+) T cell function. PMID- 12646623 TI - Role of MHC class I in immune surveillance of mitochondrial DNA integrity. AB - Mitochondrial DNA is subject to increased rates of mutations due to its proximity to the source of reactive oxygen species. Here we show that increased MHC class I (MHC I) expression serves to alert the immune system to cells with mitochondrial mutations. MHC I is overexpressed in fibroblasts with mitochondrial dysfunction from patients with mitochondrial encephalomyopathy, lactic acidosis, and stroke like episodes and in lymphocytes from purine nucleoside phosphorylase-deficient immune-deficient mice with mitochondrial DNA deletions. Consistent with a role of MHC I in the elimination of cells containing mitochondrial DNA mutations, mice deficient in MHC I accumulate mitochondrial DNA deletions in various tissues. These observations in both mice and humans suggest a role for the immune system in preventing reversion of mitochondrial DNA back into a parasitic state following deleterious mutations affecting mitochondrial oxidative phosphorylation. PMID- 12646624 TI - Requirement of species-specific interactions for the activation of human gamma delta T cells by pamidronate. AB - Human gammadelta T cells bearing Vgamma2Vdelta2-TCR recognize various kinds of small nonpeptide Ags, and activation of them by a nitrogen-containing bisphosphonate Ag, pamidronate, requires Ag presentation by cells other than gammadelta T cells, including many human tumor cells. Present results demonstrated that tumor cell lines of nonhuman origins pulsed with pamidronate failed to activate human gammadelta T cells without exception, whereas most if not all human tumor cell lines could do so. Gammadelta T cells formed stable conjugates with pamidronate-pulsed human tumor cells and both conjugate formation and gammadelta T cell activation were inhibited significantly by anti-LFA-1 mAb, suggesting the requirement of LFA-1-mediated interaction with APC for efficient gammadelta T cell activation. Consistently, ICAM-1(low) tumor cell lines pulsed with pamidronate induced no or only weak activation of gammadelta T cells, whereas similarly treated ICAM-1(high) cell lines could activate them. One of the two ICAM-1(low) tumor cell lines pulsed with pamidronate induced strong gammadelta T cell activation after ICAM-1 gene transfer. However, another ICAM 1(low) human cell line as well as murine tumor cell lines pulsed with pamidronate remained totally defective in gammadelta T cell activation even after expression of human ICAM-1. These results suggested that activation of human gammadelta T cells by nonpeptide Ags required species-specific interactions in addition to LFA 1/ICAM-1-mediated cell adhesion with APC. PMID- 12646625 TI - The molecular basis for the lack of immunostimulatory activity of vertebrate DNA. AB - Macrophages and B cells are activated by unmethylated CpG-containing sequences in bacterial DNA. The lack of activity of self DNA has generally been attributed to CpG suppression and methylation, although the role of methylation is in doubt. The frequency of CpG in the mouse genome is 12.5% of Escherichia coli, with unmethylated CpG occurring at approximately 3% the frequency of E. coli. This suppression of CpG alone is insufficient to explain the inactivity of self DNA; vertebrate DNA was inactive at 100 micro g/ml, 3000 times the concentration at which E. coli DNA activity was observed. We sought to resolve why self DNA does not activate macrophages. Known active CpG motifs occurred in the mouse genome at 18% of random occurrence, similar to general CpG suppression. To examine the contribution of methylation, genomic DNAs were PCR amplified. Removal of methylation from the mouse genome revealed activity that was 23-fold lower than E. coli DNA, although there is only a 7-fold lower frequency of known active CpG motifs in the mouse genome. This discrepancy may be explained by G-rich sequences such as GGAGGGG, which potently inhibited activation and are found in greater frequency in the mouse than the E. coli genome. In summary, general CpG suppression, CpG methylation, inhibitory motifs, and saturable DNA uptake combined to explain the inactivity of self DNA. The immunostimulatory activity of DNA is determined by the frequency of unmethylated stimulatory sequences within an individual DNA strand and the ratio of stimulatory to inhibitory sequences. PMID- 12646626 TI - More is not necessarily better: prozone-like effects in passive immunization with IgG. AB - Despite a century of study, the relationship between Ag-specific Ig concentration and protection remains poorly understood for the majority of pathogens. In certain conditions, administration of high Ab doses before challenge with an infectious agent can be less effective than smaller Ab doses, a phenomenon which is consistent with a prozone-like effect. In this study, the relationship between IgG1, IgG2a, IgG2b, and IgG3 dose, infective inocula, and protection was investigated in a mouse model of Cryptococcus neoformans infection. The activity of each IgG subclass ranged from protective to disease-enhancing depending on both the Ab dose and infective inocula used. Enhanced dissemination to the brain was observed in mice given a high IgG2a dose and a relatively low inoculum. Ab administration had immunomodulatory effects, with cytokine expression in lung, brain, and spleen varying as a function of the infective inoculum Ab dose and IgG subclass. In vitro studies did not predict or explain the mechanism of in vivo prozone-like effects, because all isotypes were opsonic and elicited NO release from macrophages. IgG2a was most efficient in inducing a macrophage oxidative burst. These results reveal that an individual Ab can be protective, nonprotective, or disease-enhancing depending on its concentration relative to a challenge inoculum. Our findings have implications for the potential contribution of Ab responses to defense against microbial diseases because Ab-mediated immunity may be protective, nonprotective, or even deleterious to the host. PMID- 12646627 TI - Galectin-9 induces apoptosis through the calcium-calpain-caspase-1 pathway. AB - Galectin-9 (Gal-9) induced the apoptosis of not only T cell lines but also of other types of cell lines in a dose- and time-dependent manner. The apoptosis was suppressed by lactose, but not by sucrose, indicating that beta-galactoside binding is essential for Gal-9-induced apoptosis. Moreover, Gal-9 required at least 60 min of Gal-9 binding and possibly de novo protein synthesis to mediate the apoptosis. We also assessed the apoptosis of peripheral blood T cells by Gal 9. Apoptosis was induced in both activated CD4(+) and CD8(+) T cells, but the former were more susceptible than the latter. A pan-caspase inhibitor (Z-VAD-FMK) inhibited Gal-9-induced apoptosis. Furthermore, a caspase-1 inhibitor (Z-YVAD FMK), but not others such as Z-IETD-FMK (caspase-8 inhibitor), Z-LEHD-FMK (caspase-9 inhibitor), and Z-AEVD-FMK (caspase-10 inhibitor), inhibited Gal-9 induced apoptosis. We also found that a calpain inhibitor (Z-LLY-FMK) suppresses Gal-9-induced apoptosis, that Gal-9 induces calcium (Ca(2+)) influx, and that either the intracellular Ca(2+) chelator BAPTA-AM or an inositol trisphosphate inhibitor 2-aminoethoxydiphenyl borate inhibits Gal-9-induced apoptosis. These results suggest that Gal-9 induces apoptosis via the Ca(2+)-calpain-caspase-1 pathway, and that Gal-9 plays a role in immunomodulation of T cell-mediated immune responses. PMID- 12646628 TI - B7-H1 (programmed death-1 ligand) on dendritic cells is involved in the induction and maintenance of T cell anergy. AB - In an effort to identify immunoregulatory molecules on dendritic cells (DC), we generated and screened for mAbs capable of modulating the T cell stimulatory function of DC. A particularly interesting mAb was mAb DF272. It recognizes monocyte-derived DC, but not blood monocytes or lymphocytes, and has profound immunomodulatory effects on DC. Treatment of DC with intact IgG or Fab of mAb DF272 enhanced their T cell stimulatory capacity. This effect on DC was accompanied by neither an up-regulation of costimulatory molecules such as B7.1 (CD80), B7.2 (CD86), and MHC class II molecules nor by an induction of cytokine production, including IL-1, TNF-alpha, IL-10, and IL-12. Moreover, the well established inhibitory function of IL-10-treated DC could be reverted with mAb DF272. Even T cells, anergized because of stimulation with IL-10-treated DC, could be reactivated and induced to proliferate upon stimulation with mAb DF272 treated DC. Furthermore, mAb DF272-treated DC favored the induction of a type-1 cytokine response in T cells and inhibited IL-10 production. By using a retrovirus-based cDNA expression library generated from DC, we cloned and sequenced the mAb DF272-defined cell surface receptor and could demonstrate that it is identical with B7-H1 (programmed death-1 ligand), a recently identified new member of the B7 family of costimulatory molecules. Our results thus demonstrate that the mAb DF272-defined surface molecule B7-H1 represents a unique receptor structure on DC that might play a role in the induction and maintenance of T cell anergy. PMID- 12646630 TI - SS-A/Ro52, an autoantigen involved in CD28-mediated IL-2 production. AB - An autoantibody against SS-A/Ro52 (Ro52) is most frequently found in the sera of patients with Sjogren's syndrome, systemic lupus erythematosus, and congenital heart block from anti-Ro52 Ab-positive mother. However, the physiological function of the autoantigen SS-A/Ro52 has not yet been elucidated. In this study, we describe the role of Ro52 protein in T cell activation. Overexpression of SS A/Ro52 in Jurkat T cell resulted in enhanced IL-2 production following CD28 stimulation. Furthermore, transfection of anti-Ro52-specific small RNA duplexes partially blocked the expression of native and overexpressed Ro52 in Jurkat T cell, resulting in decreased IL-2 production via CD28 pathway in these cells. Finally, intracellular localization of Ro52 dramatically changed following CD28 stimulation. Our data reveal a novel function of Ro52 in CD28-mediated pathway, which eventually contributes to cytokine production and expression of the T cell biological programs. PMID- 12646631 TI - Sphingosine-1-phosphate receptor agonism impairs the efficiency of the local immune response by altering trafficking of naive and antigen-activated CD4+ T cells. AB - FTY720 (2-amino-[2-(4-octylphenyl) ethyl]-1,3-propanediol hydrochloride) is an immunosuppressive agent that inhibits allograft rejection. We recently demonstrated that FTY-phosphate, the active metabolite of FTY720, acts as a full agonist for sphingosine-1-phosphate (S1P) receptors. Furthermore, activation of S1P receptors with their natural ligand, S1P, as well as pharmacological ligands leads to lymphopenia, probably due to sequestration of lymphocytes in secondary lymphoid organs. In the present study we used a local Ag-challenged mouse model to examine the effects of FTY720 on T cell activation in the draining lymph node (DLN) and on the release of activated T cells to the peripheral blood compartment. We showed that the number of Ag-activated CD4(+) T cells in the DLN after injection of Ag and CFA into a footpad was dramatically reduced after FTY720 treatment. However, T cell proliferation, both in vitro and in vivo, was not impaired by FTY720. Our results suggest that the reduced efficiency of T cell responses in the DLN in response to a local Ag is probably due to a defective recirculation of naive T cells caused by FTY720 treatment. Furthermore, we found that the numbers of naive and Ag-activated CD4(+) T cells in the peripheral blood of Ag-challenged mice were equally reduced with FTY720 treatment, suggesting that both T cell subsets are sequestered in the DLNs. Thus, FTY720 induces immunosuppression through inhibition of both the recirculation of naive T cells and the release of Ag-activated T cells from the DLN to lymph and to the blood compartment. PMID- 12646632 TI - B cell complement receptor 2 transfer reaction. AB - The B cell C receptor specific for C3dg (CR2) shares a number of features with the primate E C receptor (CR1). Previously, we have demonstrated, both in vitro and in animal models, that immune complexes (IC) bound to primate E CR1, either via C opsonization or by means of bispecific mAb complexes, can be transferred to acceptor macrophages in a process that also removes CR1 from the E. We have now extended this paradigm, the transfer reaction, to include B cell CR2. We used both flow cytometry and fluorescence microscopy to demonstrate that IC bound to Raji cell CR2, either via C opsonization or through the use of an anti-CR2 mAb, are transferred to acceptor THP-1 cells. This reaction, which appears to require Fc recognition of IgG bound to Raji cell CR2, also leads to transfer of CR2. Additional support for the B cell transfer reaction is provided in a prototype study in a monkey model in which IC bound to B cell CR2 are localized to the spleen. These findings may have important implications with respect to defining the role of C in IC handling during the normal immune response. PMID- 12646629 TI - TGF beta 1 inhibits Ca2+-calcineurin-mediated activation in thymocytes. AB - TGFbeta1 is a polypeptide growth modulatory and differentiation factor involved in many biological processes including immune homeostasis and self-tolerance. Tgfb1 knockout mice die around weaning age due to severe inflammation in most major organ systems, but the mechanism underlying this disease is not understood. In this study we demonstrate that Tgfb1(-/-) CD4(+)CD8(+) and CD4(+)CD8(-) thymocytes are hyperresponsive to receptor-mediated and receptor-independent mitogenic stimulation. A suboptimal concentration of ionomycin in the presence of PMA fully activates Tgfb1(-/-) thymocytes, whereas the inhibitors of Ca(2+) influx and calcineurin, EGTA and FK506, eliminate the hyperresponsiveness. Hence, the hypersensitivity of Tgfb1(-/-) thymocytes is due to a lowered threshold for Ca(2+)-dependent activation. Further, we demonstrate that the hypersensitivity of thymocytes results from the absence of TGFbeta1 and not from the inflammatory environment because the thymocytes are hyperresponsive in preinflammatory-stage Tgfb1(-/-) mice. Our results suggest for the first time that TGFbeta1 functions to inhibit aberrant T cell expansion by maintaining intracellular calcium concentration levels low enough to prevent a mitogenic response by Ca(2+) independent stimulatory pathways alone. Consequently, TGFbeta1 prevents autoimmune disease through a Ca(2+) regulatory pathway that maintains the activation threshold above that inducible by self-MHC-TCR interactions. PMID- 12646634 TI - Lipopolysaccharide promotes the survival of osteoclasts via Toll-like receptor 4, but cytokine production of osteoclasts in response to lipopolysaccharide is different from that of macrophages. AB - Lipopolysaccharide is a pathogen that causes inflammatory bone loss. Monocytes and macrophages produce proinflammatory cytokines such as IL-1, TNF-alpha, and IL 6 in response to LPS. We examined the effects of LPS on the function of osteoclasts formed in vitro in comparison with its effect on bone marrow macrophages, osteoclast precursors. Both osteoclasts and bone marrow macrophages expressed mRNA of Toll-like receptor 4 (TLR4) and CD14, components of the LPS receptor system. LPS induced rapid degradation of I-kappaB in osteoclasts, and stimulated the survival of osteoclasts. LPS failed to support the survival of osteoclasts derived from C3H/HeJ mice, which possess a missense mutation in the TLR4 gene. The LPS-promoted survival of osteoclasts was not mediated by any of the cytokines known to prolong the survival of osteoclasts, such as IL-1beta, TNF alpha, and receptor activator of NF-kappaB ligand. LPS stimulated the production of proinflammatory cytokines such as IL-1beta, TNF-alpha, and IL-6 in bone marrow macrophages and peritoneal macrophages, but not in osteoclasts. These results indicate that osteoclasts respond to LPS through TLR4, but the characteristics of osteoclasts are quite different from those of their precursors, macrophages, in terms of proinflammatory cytokine production in response to LPS. PMID- 12646633 TI - The possible role of TGF-beta-induced suppressors of cytokine signaling expression in osteoclast/macrophage lineage commitment in vitro. AB - Osteoclast formation is dependent on the ability of TGF-beta to enable receptor activator of NF-kappaB ligand (RANKL)-induced commitment of hemopoietic precursors to the osteoclastic lineage. The mechanism by which TGF-beta enables formation is unknown. One possibility is that TGF-beta opposes Janus kinase (JAK)/STAT signals generated by inhibitory cytokines such as IFN-beta. The JAK/STAT pathway is activated by cytokines that induce resistance to osteoclast formation, such as IFN-gamma and M-CSF, and the effect of these is opposed by TGF beta. Recently, a group of STAT-induced factors, termed suppressors of cytokine signaling (SOCS), has been identified that inhibit JAK/STAT signals. Therefore, we tested the ability of TGF-beta to induce SOCS expression in osteoclast precursors and examined the effect of SOCS expression on osteoclast/macrophage lineage commitment. We found that while SOCS mRNA is undetectable in macrophages, osteoclasts express SOCS-3, and TGF-beta up-regulates this expression. Furthermore, TGF-beta rapidly induces sustained SOCS-3 expression in macrophage/osteoclast precursors. To determine whether SOCS-3 plays a role in osteoclast differentiation we expressed SOCS-3 in precursors using a retroviral system. We found that osteoclast differentiation was significantly enhanced in SOCS-3-infected precursors, and SOCS-3 expression enables formation in the presence of anti-TGF-beta Ab. On the other hand, antisense knockdown of SOCS-3 strongly suppressed osteoclast formation and significantly blunted the response to TGF-beta. Moreover, like TGF-beta, SOCS-3 expression opposed the inhibitory effect of IFN-beta. These data suggest that TGF-beta-induced expression of SOCS-3 may represent a mechanism by which TGF-beta suppresses inhibitory cytokine signaling, priming precursors for a role in bone resorption. PMID- 12646635 TI - Conservation of nonpeptide antigen recognition by rhesus monkey V gamma 2V delta 2 T cells. AB - We have previously found that monkey Vgamma2Vdelta2(+) T cells mount adaptive immune responses in response to Mycobacterium bovis bacillus Calmette-Guerin infections. We have now analyzed rhesus monkey gammadelta T cell responses to nonpeptide Ags and superantigens. Like human Vgamma2Vdelta2(+) T cells, rhesus monkey gammadelta T cells are stimulated when exposed to prenyl pyrophosphate, bisphosphonate, and alkylamine Ags. Responsiveness was limited to gammadelta T cells expressing Vgamma2Vdelta2 TCRs. Rhesus monkey Vgamma2Vdelta2(+) T cells also responded to the superantigen, staphyloccocal enterotoxin A. Sequencing of the rhesus monkey Vgamma2Vdelta2 TCR revealed a strong sequence homology to human Vgamma2Vdelta2 TCR that preserves important sequence motifs. Moreover, chimeric TCRs that pair human Vgamma2 with monkey Vdelta2 and monkey Vgamma2 with human Vdelta2 retain reactivity to nonpeptide Ags and B cell lymphomas. A molecular model of the rhesus monkey Vgamma2Vdelta2 TCR has a basic region in the complementarity-determining region 3 binding groove that is similar to that seen in the human Vgamma2Vdelta2 TCR and preserves the topology of the complementarity determining region loops. Thus, recognition of nonpeptide prenyl pyrophosphate, bisphosphonate, and alkylamine Ags is conserved in primates suggesting that primates can provide an animal model for human gammadelta T cell Ag responses. PMID- 12646636 TI - ATM is not required in somatic hypermutation of VH, but is involved in the introduction of mutations in the switch mu region. AB - Class switch recombination (CSR) and somatic hypermutation (SHM) are mechanistically related processes that share common key factors such as activation-induced cytidine deaminase. We have previously shown a role for ATM (mutated in ataxia-telangiectasia) in CSR. In this paper we show that the frequency, distribution, and nature of base pair substitutions in the Ig variable (V) heavy chain genes in ataxia-telangiectasia patients are largely similar to those in normal donors, suggesting a normal SHM process. Characterization of the third complementarity-determining region in B cells from ataxia-telangiectasia patients also shows a normal V(D)J recombination process. SHM-like mutations could be identified in the switch (S) mu region (up to several hundred base pairs upstream of the S mu -S(alpha) breakpoints) in normal in vivo switched human B cells. In the absence of ATM, mutations can still be found in this region, but at less than half the frequency of that in normal donors. The latter mutations are mainly due to transitions (86% compared with 58% in controls) and are biased to A or T nucleotides. An ATM-dependent mechanism, different from that generating SHM in V genes, is therefore likely to be involved in introducing SHM-like mutations in the S region. ATM may thus be one of the factors that is not shared by the CSR and SHM processes. PMID- 12646637 TI - Transactivation of gp130 in myeloma cells. AB - Receptor transactivation, i.e., interaction between unrelated receptor systems, is a growing theme in cytokine and growth factor signaling. In this study we reveal for the first time the ability of IFN-alpha to transactivate gp130 in myeloma cells. An epidermal growth factor receptor/gp130 chimeric receptor previously shown by us to transactivate endogenous gp130, provided a complementary tool to study the underlying mechanisms of receptor cross-talk. Further analysis revealed that transactivation of gp130 by IFN-alpha did not require the extracellular or trans-membrane domain of gp130. Moreover, transactivation of gp130 was critically dependent upon Janus kinase activation by the initiating receptor and correlated with rapid and sustained Janus kinase 1 and tyrosine kinase (Tyk) 2 tyrosine phosphorylation. Finally, transactivation of gp130 may be a common theme in myeloma cells, perhaps providing a mechanism for enhanced or qualitatively distinct cellular responses to specific stimuli. PMID- 12646638 TI - c-Rel is required for chromatin remodeling across the IL-2 gene promoter. AB - IL-2 gene transcription occurs in an activation-dependent manner in T cells responding to TCR and CD28 activation. One of the critical events leading to increased IL-2 transcription is an alteration in chromatin structure across the 300-bp promoter region of the gene. We initially showed that IL-2 gene transcription in CD4(+) primary T cells is dependent on the NF-kappaB family member, c-Rel, but not RelA. We found that c-Rel is essential for global changes in chromatin structure across the 300-bp IL-2 promoter in response to CD3/CD28 in primary CD4(+) T cells, but not in response to pharmacological signals, paralleling the requirement for c-Rel in IL-2 mRNA and protein accumulation. Interestingly, measurement of activation-induced localized accessibility changes using restriction enzyme digestion revealed that accessibility close to the c-Rel binding site in the CD28RR region of the promoter is specifically dependent on c Rel. In contrast, restriction enzyme sites located at a distance from the CD28RR behave independently of c-Rel. These results suggest a nonredundant role for c Rel in generating a correctly remodeled chromatin state across the IL-2 promoter and imply that the strength of the signal determines the requirement for c-Rel. PMID- 12646639 TI - Regulation of the human Fc epsilon RI alpha-chain distal promoter. AB - The alpha-chain of the high affinity receptor for IgE (Fc epsilon RI) is essential for cell surface expression of Fc epsilon RI and binding of the IgE Ab. The human alpha-chain gene possesses two promoters: the proximal promoter, which is highly conserved with that of rodent; and the distal promoter, the structure and role of which are largely unknown. Transcriptional regulation of the alpha chain distal promoter was investigated in this study. Transient reporter assay revealed critical region for transcription activity located within -27/-17. EMSA identified Elf-1, YY1, and PU.1 as transcription factors binding to this region. In contrast to the proximal promoter, which was trans-activated by YY1 and PU.1, these transcription factors exhibited repressive function on this promoter. Addition of IL-4 caused a marked increase in transcription from the distal promoter and subsequently increased the intracellular production of the alpha chain. These results indicate that IL-4-dependent up-regulation of the human alpha-chain was due to enhancement of distal promoter activity and suggests that the two promoters have different regulatory mechanisms for alpha-chain expression. PMID- 12646640 TI - A new model of sheep Ig diversification: shifting the emphasis toward combinatorial mechanisms and away from hypermutation. AB - The current model of Ig repertoire development in sheep focuses on the rearrangement of a small number (approximately 20) of Vlambda gene segments. It is believed that this limited combinatorial repertoire is then further diversified through postrearrangement somatic hypermutation. This process has been reported to introduce as many as 110 mutations/1000 nucleotides. In contrast, our data have that indicated somatic hypermutation may diversify the preimmune repertoire to a much lesser extent. We have identified 64 new Vlambda gene segments within the rearranged Ig repertoire. As a result, many of the unique nucleotide patterns thought to be the product of somatic hypermutation are actually hard-coded within the germline. We suggest that combinatorial rearrangement makes a much larger contribution, and somatic hypermutation makes a much smaller contribution to the generation of diversity within the sheep Ig repertoire than is currently acknowledged. PMID- 12646641 TI - DM loss in k haplotype mice reveals isotype-specific chaperone requirements. AB - DM actions as a class II chaperone promote capture of diverse peptides inside the endocytic compartment(s). DM mutant cells studied to date express class II bound by class II-associated invariant chain-derived peptide (CLIP), a short proteolytic fragment of the invariant chain, and exhibit defective peptide loading abilities. To evaluate DM functional contributions in k haplotype mice, we engineered a novel mutation at the DMa locus via embryonic stem cell technology. The present experiments demonstrate short-lived A(k)/CLIP complexes, decreased A(k) surface expression, and enhanced A(k) peptide binding activities. Thus, we conclude that DM loss in k haplotype mice creates a substantial pool of empty or loosely occupied A(k) conformers. On the other hand, the mutation hardly affects E(k) activities. The appearance of mature compact E(k) dimers, near normal surface expression, and efficient Ag presentation capabilities strengthen the evidence for isotype-specific DM requirements. In contrast to DM mutants described previously, partial occupancy by wild-type ligands is sufficient to eliminate antiself reactivity. Mass spectrometry profiles reveal A(k)/CLIP and a heterogeneous collection of relatively short peptides bound to E(k) molecules. These experiments demonstrate that DM has distinct roles depending on its specific class II partners. PMID- 12646642 TI - Actin tyrosine dephosphorylation by the Src homology 1-containing protein tyrosine phosphatase is essential for actin depolymerization after membrane IgM cross-linking. AB - Src homology protein 1 (SHP-1) plays an important role in B cell Ag receptor (BCR) differentiation, proliferation, survival, and apoptosis. After BCR stimulation in apoptotic cells, SHP-1 has been shown to be recruited to phosphorylated immunoreceptor tyrosine-based inhibitory motifs present in receptors such as CD22 and CD72. However, the substrates of SHP-1 in the chicken B cell line, DT40, have remained undefined. To identify SHP-1 substrates in DT40, we used a trapping mutant, SHP-1 C/S (a catalytically inactive form). Cross linking of BCR induced hyperphosphorylation of approximately 44-kDa protein in C/S transfectants. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis revealed that this was actin (cytoplasmic type 5) carrying three immunoreceptor tyrosine-based inhibitory motif-like sequences. SHP 1 was shown to bind to one of these sequences in synthetic peptide binding experiment. Thus, actin is a direct SHP-1 substrate. Furthermore, more SHP-1 molecules translocate into lipid rafts, and their association with actin was increased after BCR stimulation. In C/S transfectants, actin polymerization induced by membrane IgM ligation was sustained to a greater extent for a longer time compared with wild-type transfectants. Therefore, actin dephosphorylation by SHP-1 is essential for actin depolymerization after BCR stimulation. Our data suggest that SHP-1 plays a pivotal role in reorganization of cytoskeletal architecture inducing actin dephosphorylation. These results clearly demonstrate the direct interaction of SHP-1 with actin. PMID- 12646643 TI - Investigation of early events in Fc epsilon RI-mediated signaling using a detailed mathematical model. AB - Aggregation of Fc epsilon RI on mast cells and basophils leads to autophosphorylation and increased activity of the cytosolic protein tyrosine kinase Syk. We investigated the roles of the Src kinase Lyn, the immunoreceptor tyrosine-based activation motifs (ITAMs) on the beta and gamma subunits of Fc epsilon RI, and Syk itself in the activation of Syk. Our approach was to build a detailed mathematical model of reactions involving Fc epsilon RI, Lyn, Syk, and a bivalent ligand that aggregates Fc(epsilon)RI. We applied the model to experiments in which covalently cross-linked IgE dimers stimulate rat basophilic leukemia cells. The model makes it possible to test the consistency of mechanistic assumptions with data that alone provide limited mechanistic insight. For example, the model helps sort out mechanisms that jointly control dephosphorylation of receptor subunits. In addition, interpreted in the context of the model, experimentally observed differences between the beta- and gamma chains with respect to levels of phosphorylation and rates of dephosphorylation indicate that most cellular Syk, but only a small fraction of Lyn, is available to interact with receptors. We also show that although the beta ITAM acts to amplify signaling in experimental systems where its role has been investigated, there are conditions under which the beta ITAM will act as an inhibitor. PMID- 12646644 TI - Depletion of neutrophils in IL-10(-/-) mice delays clearance of gastric Helicobacter infection and decreases the Th1 immune response to Helicobacter. AB - Gastric infection with Helicobacter induces a lymphocyte-rich mucosal inflammation that contains a minor population of neutrophilic granulocytes. The function of neutrophils in the local immune response to gastric Helicobacter infection remains unknown. To investigate this issue, we conducted experiments in neutrophil-depleted control wild-type (wt) and IL-10(-/-) mice infected with Helicobacter felis by gastric lavage. Infection of wt mice elicited a mild, focal gastritis and a Helicobacter-specific Th1 immune response. In wt mice Helicobacter colonization of the stomach was persistent and progressively increased during the 29 days of observation. Infection of IL-10(-/-) mice with H. felis elicited a severe chronic gastritis and a greatly enhanced Helicobacter specific Th1 immune response, as compared with wt mice. After initial colonization, the IL-0(-/-) mice completely cleared Helicobacter from the stomach by day 8. The gastric inflammation in wt and IL-10(-/-) mice contained modest numbers of neutrophils. The intensity of gastric inflammation and the extent of Helicobacter colonization were similar in control and in neutrophil-depleted wt mice. In contrast, neutrophil depletion of Helicobacter-infected IL-10(-/-) mice decreased the severity of gastritis, modulated the Helicobacter-specific Th1 immune response, and delayed the clearance of bacteria from the stomach. These studies identify a role for neutrophils in the local and systemic immune response to gastric Helicobacter in IL-10(-/-) mice. PMID- 12646645 TI - The hypervariable region of Anaplasma marginale major surface protein 2 (MSP2) contains multiple immunodominant CD4+ T lymphocyte epitopes that elicit variant specific proliferative and IFN-gamma responses in MSP2 vaccinates. AB - Major surface protein 2 (MSP2) is an immunodominant outer membrane protein of Anaplasma marginale and Anaplasma phagocytophilum pathogens that cause bovine anaplasmosis and human granulocytic ehrlichiosis, respectively. MSP2 has a central hypervariable region (HVR) flanked by highly conserved amino and carboxyl termini. During A. marginale infection, dynamic and extensive amino acid sequence variation in MSP2 occurs through recombination of msp2 pseudogenes into the msp2 expression site, followed by sequential segmental gene conversions to generate additional variants. We hypothesized that MSP2 variation leads to significant changes in Th cell recognition of epitopes in the HVR. T cell epitopes were mapped using T cells from native MSP2-immunized cattle and overlapping peptides spanning the most abundant of five different MSP2 HVRs in the immunogen. Several epitopes elicited potent effector/memory Th cell proliferative and IFN-gamma responses, including those in three discreet blocks of sequence that undergo segmental gene conversion. Th cell clones specific for an epitope in the block 1 region of the predominant MSP2 variant type failed to respond to naturally occurring variants. However, some of these variants were recognized by oligoclonal T cell lines from MSP2 vaccinates, indicating that the variant sequences contain immunogenic CD4(+) T cell epitopes. In competition/antagonism assays, the nonstimulatory variants were not inhibitory for CD4(+) T cells specific for the agonist peptide. Dynamic amino acid sequence variation in MSP2 results in escape from recognition by some effector/memory MSP2-specific Th cells. Antigenic variation in MSP2 Th cell and B cell epitopes may contribute to immune evasion that allows long-term persistence of A. marginale in the mammalian reservoir. PMID- 12646646 TI - A common mucosal chemokine (mucosae-associated epithelial chemokine/CCL28) selectively attracts IgA plasmablasts. AB - IgA immunoblasts can seed both intestinal and nonintestinal mucosal sites following localized mucosal immunization, an observation that has led to the concept of a common mucosal immune system. In this study, we demonstrate that the mucosae-associated epithelial chemokine, MEC (CCL28), which is expressed by epithelia in diverse mucosal tissues, is selectively chemotactic for IgA Ab secreting cells (ASC): MEC attracts IgA- but not IgG- or IgM-producing ASC from both intestinal and nonintestinal lymphoid and effector tissues, including the intestines, lungs, and lymph nodes draining the bronchopulmonary tree and oral cavity. In contrast, the small intestinal chemokine, TECK (CCL25), attracts an overlapping subpopulation of IgA ASC concentrated in the small intestines and its draining lymphoid tissues. Surprisingly, T cells from mucosal sites fail to respond to MEC. These findings suggest a broad and unifying role for MEC in the physiology of the mucosal IgA immune system. PMID- 12646647 TI - Tumor-derived TGF-beta reduces the efficacy of dendritic cell/tumor fusion vaccine. AB - Dendritic cell (DC)-based antitumor vaccine is a novel cancer immunotherapy that is promising for reducing cancer-related mortality. However, results from early clinical trials were suboptimal. A possible explanation is that many tumors secrete immunosuppressive factors such as TGF-beta, which may hamper host immune response to DC vaccine. In this study, we demonstrated that TGF-beta produced by tumors significantly reduced the potency of DC/tumor fusion vaccines. TGF-beta secreting (CT26-TGF-beta) stable mouse colon cancer cell lines were generated using a retroviral vector expressing TGF-beta. A non-TGF-beta-secreting (CT26 neo) cell line was generated using an empty retroviral vector. The efficacies of DC/tumor fusion vaccines were assessed in vitro and in vivo. DC/CT26-TGF-beta fusion cells failed to induce a strong T cell proliferative response in vitro, mainly due to the effect of TGF-beta on T cell responsiveness rather than DC stimulatory capability. Animals vaccinated with DC/CT26-TGF-beta fusion vaccine had lower tumor-specific CTL activity and had significantly lower survival after tumor challenge as compared with animals immunized with DC/CT26-neo hybrids (45 vs 77%, p < 0.05). Ex vivo exposure of DCs to TGF-beta did not appear to lessen the efficacy of DC vaccine. These data suggest that tumor-derived TGF-beta reduces the efficacy of DC/tumor fusion vaccine via an in vivo mechanism. Neutralization of TGF-beta produced by the fusion cells may enhance the effectiveness of DC-based immunotherapy. PMID- 12646648 TI - A role for triggering receptor expressed on myeloid cells-1 in host defense during the early-induced and adaptive phases of the immune response. AB - Triggering receptor expressed on myeloid cells (TREM)-1 is a cell surface molecule expressed on neutrophils and monocytes implicated in the propagation of the inflammatory response. To further characterize the function of this molecule in different phases of the immune response, we examined TREM-1 in the context of host defense against microbial pathogens. In primary human monocytes TREM-1 activation did not trigger innate antimicrobial pathways directed against intracellular Mycobacterium tuberculosis, and only minimally improved phagocytosis. However, activation of TREM-1 on monocytes did drive robust production of proinflammatory chemokines such as macrophage inflammatory protein 1alpha and IL-8. Engagement of TREM-1 in combination with microbial ligands that activate Toll-like receptors also synergistically increased production of the proinflammatory cytokines TNF-alpha and GM-CSF, while inhibiting production of IL 10, an anti-inflammatory cytokine. Expression of TREM-1 was up-regulated in response to TLR activation, an effect further enhanced by GM-CSF and TNF-alpha but inhibited by IL-10. Functionally, primary monocytes differentiated into immature dendritic cells following activation through TREM-1, evidenced by higher expression of CD1a, CD86, and MHC class II molecules. These cells had an improved ability to elicit T cell proliferation and production of IFN-gamma. Our data suggest that activation of TREM-1 on monocytes participates during the early induced and adaptive immune responses involved in host defense against microbial challenges. PMID- 12646649 TI - The resolution of relapsing fever borreliosis requires IgM and is concurrent with expansion of B1b lymphocytes. AB - The rate of pathogen clearance is a critical determinant of morbidity and mortality. We sought to characterize the immune response responsible for the remarkably rapid clearance of individual episodes of bacteremia caused by the relapsing fever bacterium, Borrelia hermsii. SCID or Rag(-/-) mice were incapable of resolving B. hermsii infection, indicating a critical role for T and/or B cells. TCR(-/-) mice, which lack T cells, and IL-7(-/-) mice, which are deficient in both T cells and follicular B cells, but not in B1 cells and splenic marginal zone (MZ) B cells, efficiently cleared B. hermsii. These findings suggested that B1 cells and/or MZ B cells, two B cell subsets that are known to participate in rapid, T-independent responses, might be involved. The efficient resolution of the episodes of moderate level bacteremia by splenectomized mice suggested that MZ B cells do not play the primary role in clearance of this bacterium. In contrast, xid mice, which are deficient in B1 cells, suffered more severe episodes of bacteremia than wild-type mice. The hypothesis that B1 cells are critical for clearance of B. hermsii was further supported by a selective expansion of the B1b (i.e., IgM(high), IgD(-/low), Mac1(+) CD23(-), and CD5(-)) cell subset in infected xid mice, which coincided with the eventual resolution of infection. Finally, mice selectively incapable of secreting IgM, the dominant isotype produced by B1 cells, were completely unable to clear B. hermsii. Together these results support the model that B1b cells generate the T independent IgM required for the control and resolution of relapsing fever borreliosis. PMID- 12646650 TI - Differential gamma-herpesvirus distribution in distinct anatomical locations and cell subsets during persistent infection in mice. AB - Murine gamma-herpesvirus 68 (MHV-68) provides an important experimental model for analyzing gamma-herpesvirus latent infection. After intranasal infection with MHV 68, we analyzed the distribution of the virus in different anatomical locations and purified populations of cells. Our data show that long-term latency is maintained in a variety of anatomical locations and cell populations with different frequencies. Importantly, we demonstrate that although latency in the lung is established in a variety of cell subsets, long-term latency in the lung is only maintained in B cells. In contrast, splenic latency is maintained in macrophages and dendritic cells, as well as in B cells. In blood, isotype switched B cells constitute the major viral reservoir. These results show that the cell subsets in which latency is established vary within different anatomical sites. Finally, we demonstrate that long-term latency is accompanied by a low level of infectious virus in lung and spleen. These data have important implications for understanding the establishment and maintenance of latency by gamma(2)-herpesviruses. PMID- 12646651 TI - IL-4 exacerbates anaphylaxis. AB - We evaluated whether IL-4, a cytokine critical for inducing allergic responses, also contributes to the effector phase of allergy. Pretreatment of mice with IL-4 or the related cytokine, IL-13, rapidly and dramatically increased the severity of anaphylaxis induced by cross-linking Fc(epsilon)RI or FcgammaRIII. This effect was inhibited by endogenously produced IFN-gamma, was T cell-, B cell-, and common gamma-chain-independent, and required IL-4Ralpha and Stat6. IL-4Ralpha signaling also enhanced anaphylaxis in mice infected with a nematode parasite that stimulates IL-4/IL-13 production. IL-4 exacerbated anaphylaxis by acting synergistically with vasoactive mediators to increase vascular permeability. Synergy between IL-4 and vasoactive mediators during the effector phase of allergic inflammation may both contribute to allergic immunopathology and enhance protective immunity against gastrointestinal worms. PMID- 12646652 TI - Unique regulation of CCL18 production by maturing dendritic cells. AB - Dendritic cells (DC) orchestrate the trafficking of lymphocytes by secreting chemokines with different specificity and function. Chemokines are produced at higher levels by mature DC. This study shows that CCL18 is one of the most abundant chemokines produced by immature DC. In contrast to all other chemokines investigated to date, CCL18 was selectively down-regulated during the maturation process induced by LPS, TNF, CD40 ligand, Staphylococcus aureus Cowan I, Candida albicans, and influenza virus. IL-10 and vitamin D(3), two known inhibitors of DC differentiation and function, strongly promoted CCL18 secretion, whereas IFN gamma, a costimulator of DC function, inhibited its production. IL-10 also induced CCL18 secretion in blood myeloid DC. No CCL18 secretion was observed in blood plasmacytoid DC. The opposite pattern of regulation was observed for CCL20, a prototypic inflammatory chemokine. CCL18 was found to be a chemotactic factor for immature DC. Therefore, CCL18 may act as a chemotactic signal that promotes the colocalization of immature DC with naive T lymphocytes in an IL-10-dominated environment with the consequent generation of T regulatory cells. These characteristics suggest that CCL18 may be part of an inhibitory pathway devoted to limiting the generation of specific immune responses at peripheral sites. PMID- 12646653 TI - The extracellular matrix and cytokines regulate microglial integrin expression and activation. AB - Microglia are the primary immune effector cells resident within the CNS, whose activation into migratory, phagocytic cells is associated with increased expression of cell adhesion molecules of the integrin family. To determine which specific factors are important regulators of microglial activation and integrin expression, we have examined the influence of individual cytokines and extracellular matrix (ECM) substrates by quantifying cell surface expression of MHC and individual integrins by flow cytometry. We found that the proinflammatory cytokines TNF and IFN-alpha promoted microglial activation, as assessed by amoeboid morphology and increased expression of MHC class I, and also increased expression of the alpha(4)beta(1) and Mac-1 integrins. In contrast, TGF-beta1 had the opposite effect and was dominant over the other cytokines. Furthermore, the ECM substrates fibronectin and vitronectin, but not laminin, also promoted microglial activation and increased expression of the alpha(4)beta(1), alpha(5)beta(1) and Mac-1 integrins, but significantly, the influence of fibronectin and vitronectin was not diminished by TGF-beta1. Taken together, this work suggests that, in addition to cytokines, the ECM represents an important regulatory influence on microglial activity. Specifically, it implies that increases in the local availability of fibronectin or vitronectin, as a result of blood-brain barrier breakdown or increased expression in different pathological states of the CNS, could induce microglial activation and increased expression of integrins. PMID- 12646654 TI - STAT4 signal pathways regulate inflammation and airway physiology changes in allergic airway inflammation locally via alteration of chemokines. AB - Mice homozygous for the STAT4-null mutation were sensitized to cockroach Ag, challenged intratracheally 21 days later, and compared with STAT4-competent allergic mice. The STAT4(-/-) mice showed significant decreases in airway hyperreactivity (AHR) and peribronchial eosinophils compared with wild-type controls. In addition, pulmonary levels of chemokines were decreased in the STAT4(-/-) mice, including CC chemokine ligand (CCL)5, CCL6, CCL11, and CCL17. However, levels of Th2-type cytokines, such as IL-4 and IL-13, as well as serum IgE levels were similar in the two groups. Transfer of splenic lymphocytes from sensitized wild-type mice into sensitized STAT4(-/-) mice did not restore AHR in the mutant mice. Furthermore, chemokine production and peribronchial eosinophilia were not restored during the cellular transfer experiments. Thus, it appears that STAT4 expression contributes to a type 2 process such as allergen-induced chemokine production and AHR. In additional studies, competent allergic mice were treated with anti-IL-12 locally in the airways at the time of allergen rechallenge. These latter studies also demonstrated a decrease in AHR. Altogether, these data suggest that STAT4-mediated pathways play a role locally within the airway for the exacerbation of the allergen-induced responses. PMID- 12646655 TI - Modulation of cutaneous inflammation by angiotensin-converting enzyme. AB - Cutaneous neurogenic inflammation is a complex biological response of the host immune system to noxious stimuli. Present evidence suggests that zinc metalloproteases may play an important role in the regulation of neurogenic inflammation by controlling the local availability of neuropeptides, such as substance P (SP), that are capable of initiating or amplifying cutaneous inflammation after release from sensory nerves. To address the hypothesis that the dipeptidyl carboxypeptidase angiotensin-converting enzyme (ACE) is capable of modulating skin inflammation, we have analyzed murine allergic contact dermatitis (ACD) and irritant contact dermatitis (ICD) using wild-type C57BL/6J (ACE(+/+)) or genetically engineered mice with a heterozygous deletion of somatic ACE (ACE(+/-)). In 2,4-dinitro-1-fluorobenzene-sensitized ACE(+/-) mice, ACD was significantly augmented in comparison to ACE(+/+) controls as determined by the degree of ear swelling after exposure to hapten. Likewise, systemic treatment of ACE(+/+) mice with the ACE inhibitor captopril before sensitization or elicitation of ACD significantly augmented the ACD response. In contrast, local damage and neuropeptide depletion of sensory nerves following capsaicin, injection of a bradykinin B(2), or a SP receptor antagonist before sensitization significantly inhibited the augmented effector phase of ACD in mice with functionally absent ACE. However, in contrast to ACD, the response to the irritant croton oil was not significantly altered in ACE(+/-) compared with ACE(+/+) mice. Thus, ACE by degrading bradykinin and SP significantly controls cutaneous inflammatory responses to allergens but not to irritants, which may explain the frequently observed exacerbation of inflammatory skin disease in patients under medication with ACE inhibitors. PMID- 12646656 TI - Sodium phenylacetate inhibits adoptive transfer of experimental allergic encephalomyelitis in SJL/J mice at multiple steps. AB - Experimental allergic encephalomyelitis (EAE) is the animal model for multiple sclerosis. The present study underlines the importance of sodium phenylacetate (NaPA), a drug approved for urea cycle disorders, in inhibiting the disease process of adoptively transferred EAE in female SJL/J mice at multiple steps. Myelin basic protein (MBP)-primed T cells alone induced the expression of NO synthase (iNOS) and the activation of NF-kappaB in mouse microglial cells through cell-cell contact. However, pretreatment of MBP-primed T cells with NaPA markedly inhibited its ability to induce microglial expression of iNOS and activation of NF-kappaB. Consistently, adoptive transfer of MBP-primed T cells, but not that of NaPA-pretreated MBP-primed T cells, induced the clinical symptoms of EAE in female SJL/J mice. Furthermore, MBP-primed T cells isolated from NaPA-treated donor mice were also less efficient than MBP-primed T cells isolated from normal donor mice in inducing iNOS in microglial cells and transferring EAE to recipient mice. Interestingly, clinical symptoms of EAE were much less in mice receiving NaPA through drinking water than those without NaPA. Similar to NaPA, sodium phenylbutyrate, a chemically synthesized precursor of NaPA, also inhibited the disease process of EAE. Histological and immunocytochemical analysis showed that NaPA inhibited EAE-induced spinal cord mononuclear cell invasion and normalized iNOS, nitrotyrosine, and p65 (the RelA subunit of NF-kappaB) expression within the spinal cord. Taken together, our results raise the possibility that NaPA or sodium phenylbutyrate taken through drinking water or milk may reduce the observed neuroinflammation and disease process in multiple sclerosis patients. PMID- 12646657 TI - Complement factor C5a mediates renal ischemia-reperfusion injury independent from neutrophils. AB - The complement system has been shown to mediate renal ischemia-reperfusion (I/R) injury. However, the contribution of complement factor C5a to I/R injury, in particular in the kidney, remains to be established. In this study, we investigated the impact of blocking the C5aR pathway on the inflammatory response and on the renal function in a murine model of I/R injury. First, we analyzed C5aR expression in kidneys of healthy mice. Intriguingly, we found expression on mesangial, as well as on tubular epithelial, cells. After I/R injury, C5aR expression was up-regulated in tubular epithelial cells. In addition, mRNA levels of CXC chemokines and TNF-alpha increased significantly and kidneys were heavily infiltrated by neutrophils. Blocking the C5aR pathway by a specific C5a receptor antagonist (C5aRA) abrogated up-regulation of CXC chemokines but not of TNF-alpha and reduced neutrophil infiltration by >50%. Moreover, application of the C5aRA significantly reduced loss of renal function. This improvement of function was independent of the presence of neutrophils because neutrophil depletion by mAb NIMP-R14 did not affect the protective effect of C5aRA treatment. Furthermore, blocking of the C5aR pathway had no influence on renal apoptosis. These data provide evidence that C5a is crucially involved in the pathogenesis of renal I/R injury by modulation of neutrophil-dependent as well as neutrophil-independent pathways, which include the regulation of CXC chemokines but not TNF-alpha or apoptotic pathways. PMID- 12646658 TI - IFN-gamma induces high mobility group box 1 protein release partly through a TNF dependent mechanism. AB - We recently discovered that a ubiquitous protein, high mobility group box 1 protein (HMGB1), is released by activated macrophages, and functions as a late mediator of lethal systemic inflammation. To elucidate mechanisms underlying the regulation of HMGB1 release, we examined the roles of other cytokines in induction of HMGB1 release in macrophage cell cultures. Macrophage migration inhibitory factor, macrophage-inflammatory protein 1beta, and IL-6 each failed to significantly induce the release of HMGB1 even at supraphysiological levels (up to 200 ng/ml). IFN-gamma, an immunoregulatory cytokine known to mediate the innate immune response, dose-dependently induced the release of HMGB1, TNF, and NO, but not other cytokines such as IL-1alpha, IL-1beta, or IL-6. Pharmacological suppression of TNF activity with neutralizing Abs, or genetic disruption of TNF expression (TNF knockout) partially (50-60%) inhibited IFN-gamma-mediated HMGB1 release. AG490, a specific inhibitor for Janus kinase 2 of the IFN-gamma signaling pathway, dose-dependently attenuated IFN-gamma-induced HMGB1 release. These data suggest that IFN-gamma plays an important role in the regulation of HMGB1 release through a TNF- and Janus kinase 2-dependent mechanism. PMID- 12646659 TI - Intranasal immunotherapy is more effective than intradermal immunotherapy for the induction of airway allergen tolerance in Th2-sensitized mice. AB - Immunotherapy (IT) by injection more readily induces clinical tolerance to stinging insects than to respiratory allergens. However, while systemic immunization induces adaptive responses systemically, the induction of mucosal immunity generally requires local Ag exposure. Taken together, these observations suggest that the poor success rate of systemic IT for asthma could be a consequence of inadequate immune modulation in the airways. In support of this position, investigations presented in this report demonstrate that allergen IT more effectively induces airway allergen tolerance in Th2-sensitized mice, when delivered by the intranasal (i.n.) vs the intradermal (i.d.) route. Moreover, compared with native allergen, allergen immunostimulatory sequence oligodeoxynucleotide conjugate proved to be a more effective i.n. IT reagent for protecting allergic mice from airway hypersensitivity responses. Furthermore, for both native allergen and allergen immunostimulatory sequence oligodeoxynucleotide conjugate, i.n. and i.d. IT delivery were similarly effective in modulating systemic immune profiles in Th2-sensitized mice, while only i.n. IT had significant immunomodulatory activity on B and T cell responses in the airways. The present investigations may be the first to suggest that i.n. IT is more effective than i.d. IT for the treatment of asthma. Furthermore, our results suggest that modulating airway rather than systemic immunity may be the more important therapeutic target for the induction of clinical tolerance to respiratory allergens. PMID- 12646660 TI - Viremia control despite escape from a rapid and potent autologous neutralizing antibody response after therapy cessation in an HIV-1-infected individual. AB - The neutralizing Ab response after primary HIV-1 infection is delayed relative to the virus-specific CD8(+) T cell response and the initial decline in plasma viremia. Because nearly all HIV-1 infections result in AIDS, it would be instructive to study cases where neutralizing Ab production commenced sooner. This was done in subject AC10, an individual treated during early infection and in whom a rapid autologous neutralizing Ab response was detected after therapy cessation as rebound viremia declined and remained below 1000 RNA copies/ml of blood for over 2.5 years. This subject's Abs were capable of reducing the infectivity of his rebound virus by >4 logs in vitro at a time when rebound viremia was down-regulated and virus-specific CD8(+) T cells were minimal, suggesting that neutralizing Abs played an important role in the early control of viremia. The rebound virus did not exhibit an unusual phenotype that might explain its high sensitivity to neutralization by autologous sera. Neutralization escape occurred within 75 days and was proceeded by neutralizing Ab production to the escape variant and subsequent escape. Notably, escape was not associated with a significant rise in plasma viremia, perhaps due to increasing CD8(+) T cell responses. Sequence analysis of gp160 revealed a growing number of mutations over time, suggesting ongoing viral evolution in the face of potent antiviral immune responses. We postulate that an early effective neutralizing Ab response can provide long-term clinical benefits despite neutralization escape. PMID- 12646662 TI - Medical detection in the 21st century. PMID- 12646663 TI - Medicine and the racial divide. PMID- 12646661 TI - IL-12 deficiency in MRL-Fas(lpr) mice delays nephritis and intrarenal IFN-gamma expression, and diminishes systemic pathology. AB - Autoimmune disease in MRL-Fas(lpr) mice is characterized by fatal nephritis, systemic pathology, and autoantibodies, mimicking human lupus. We previously reported that 1) intrarenal IL-12 elicits nephritis by fostering the accumulation of intrarenal IFN-gamma-secreting T cells, and 2) MRL-Fas(lpr) mice deficient in the IFN-gamma receptor were spared from nephritis. Therefore, we hypothesized that eliminating IL-12 in MRL-Fas(lpr) mice reduces IFN-gamma-secreting cells and thereby prevents systemic pathology. For this purpose, we constructed an IL-12p40 deficient MRL-Fas(lpr)(IL-12(-/-)) strain. We determined that glomerular and interstitial, but not perivascular, renal pathology were decreased in IL-12(-/-) mice vs the wild-type (WT) strain (5 mo of age). Similarly, systemic pathology (lung, lacrimal and salivary glands, skin, and lymphadenopathy) was diminished. The intrarenal accumulation of T cells (CD4(+), CD8(+), CD4(-)CD8(-)B220(+)) and macrophages was dramatically reduced in IL-12(-/-) MRL-Fas(lpr) kidneys. We determined that there were fewer IFN-gamma transcripts (>70%) in the IL-12(-/-) protected kidneys compared with the WT kidneys. Similarly, cells propagated from IL-12(-/-) MRL-Fas(lpr) kidneys generated substantially less IFN-gamma when stimulated with IL-12 and IL-18 compared with those from WT kidneys, and we detected fewer CD8 and B220 T cells producing IFN-gamma in these IL-12(-/-) MRL Fas(lpr) kidneys. Of note, survival was modestly extended in the IL-12(-/-) MRL Fas(lpr) mice. While lung and lacrimal and salivary gland pathology remained reduced in moribund IL-12(-/-) MRL-Fas(lpr) mice, renal pathology and IFN-gamma expression were equivalent to those in the WT strain. Thus, we suggest that IL-12 is a therapeutic target for multiple tissues in lupus; however blocking IL-12 alone is not sufficient to confer enduring protection from lupus nephritis. PMID- 12646664 TI - A death at Duke. PMID- 12646665 TI - Clinical spectrum of obesity and mutations in the melanocortin 4 receptor gene. AB - BACKGROUND: Melanocortin 4 receptor (MC4R) deficiency is the commonest monogenic form of obesity. However, the clinical spectrum and mode of inheritance have not been defined, pathophysiological mechanisms leading to obesity are poorly understood, and there is little information regarding genotype-phenotype correlations. METHODS: We determined the nucleotide sequence of the MC4R gene in 500 probands with severe childhood obesity. Family studies were undertaken to examine cosegregation of identified mutations with obesity. Subjects with MC4R deficiency underwent metabolic and endocrine evaluation; the results were correlated with the signaling properties of mutant receptors. RESULTS: Twenty nine probands (5.8 percent) had mutations in MC4R; 23 were heterozygous, and 6 were homozygous. Mutation carriers had severe obesity, increased lean mass, increased linear growth, hyperphagia, and severe hyperinsulinemia; homozygotes were more severely affected than heterozygotes. Subjects with mutations retaining residual signaling capacity had a less severe phenotype. CONCLUSIONS: Mutations in MC4R result in a distinct obesity syndrome that is inherited in a codominant manner. Mutations leading to complete loss of function are associated with a more severe phenotype. The correlation between the signaling properties of these mutant receptors and energy intake emphasizes the key role of this receptor in the control of eating behavior in humans. PMID- 12646666 TI - Binge eating as a major phenotype of melanocortin 4 receptor gene mutations. AB - BACKGROUND: Obesity, a multifactorial disease caused by the interaction of genetic factors with the environment, is largely polygenic. A few mutations in these genes, such as in the leptin receptor (LEPR) gene and melanocortin 4 receptor (MC4R) gene, have been identified as causes of monogenic obesity. METHODS: We sequenced the complete MC4R coding region, the region of the proopiomelanocortin gene (POMC) encoding the alpha melanocyte-stimulating hormone, and the leptin-binding domain of LEPR in 469 severely obese white subjects (370 women and 99 men; mean [+/-SE] age, 41.0+/-0.5 years; body-mass index [the weight in kilograms divided by the square of the height in meters], 44.1+/-2.0). Fifteen women and 10 men without a history of dieting or a family history of obesity served as normal-weight controls (age, 47.7+/-2.0 years; body mass index, 21.6+/-0.4). Detailed phenotypic data, including information on body fat, resting energy expenditure, diet-induced thermogenesis, serum concentrations of leptin, and eating behavior, were collected. RESULTS: Twenty-four obese subjects (5.1 percent) and one control subject (4 percent) had MC4R mutations, including five novel variants. Twenty of the 24 obese subjects with an MC4R mutation were matched for age, sex, and body-mass index with 120 of the 445 obese subjects without an MC4R mutation. All mutation carriers reported binge eating, as compared with 14.2 percent of obese subjects without mutations (P<0.001) and 0 percent of the normal-weight subjects without mutations. The prevalence of binge eating was similar among carriers of mutations in the leptin-binding domain of LEPR and noncarriers. No mutations were found in the region of POMC encoding alpha melanocyte-stimulating hormone. CONCLUSIONS: Binge eating is a major phenotypic characteristic of subjects with a mutation in MC4R, a candidate gene for the control of eating behavior. PMID- 12646667 TI - Soluble CD40 ligand in acute coronary syndromes. AB - BACKGROUND: CD40 ligand is expressed on platelets and released from them on activation. We investigated the predictive value of soluble CD40 ligand as a marker for clinical outcome and the therapeutic effect of glycoprotein IIb/IIIa receptor inhibition in patients with acute coronary syndromes. METHODS: Serum levels of soluble CD40 ligand were measured in 1088 patients with acute coronary syndromes who had previously been enrolled in a randomized trial comparing abciximab with placebo before coronary angioplasty and in 626 patients with acute chest pain. RESULTS: The levels of soluble CD40 ligand were elevated (above 5.0 microg per liter) in 221 patients with acute coronary syndromes (40.6 percent). Among patients receiving placebo, elevated soluble CD40 ligand levels indicated a significantly increased risk of death or nonfatal myocardial infarction during six months of follow-up (adjusted hazard ratio as compared with patients with low levels of the ligand [< or =5.0 microg per liter], 2.71; 95 percent confidence interval, 1.51 to 5.35; P=0.001). The prognostic value of this marker was validated in the patients with chest pain, among whom elevated soluble CD40 ligand levels identified those with acute coronary syndromes who were at high risk for death or nonfatal myocardial infarction (adjusted hazard ratio as compared with those with low levels of the ligand, 6.65; 95 percent confidence interval, 3.18 to 13.89; P<0.001). The increased risk in patients with elevated soluble CD40 ligand levels was significantly reduced by treatment with abciximab (adjusted hazard ratio as compared with those receiving placebo, 0.37; 95 percent confidence interval, 0.20 to 0.68; P=0.001), whereas there was no significant treatment effect of abciximab in patients with low levels of soluble CD40 ligand. CONCLUSIONS: In patients with unstable coronary artery disease, elevation of soluble CD40 ligand levels indicated an increased risk of cardiovascular events. Elevation of soluble CD40 ligand identifies a subgroup of patients at high risk who are likely to benefit from antiplatelet treatment with abciximab. PMID- 12646668 TI - An outbreak of conjunctivitis due to atypical Streptococcus pneumoniae. AB - BACKGROUND: In February 2002, clinicians at the Dartmouth College Health Service recognized an outbreak of conjunctivitis; cultures of conjunctival swabs implicated Streptococcus pneumoniae. An investigation was begun to determine the extent of the outbreak, confirm the cause, identify modes of transmission, and implement control measures. METHODS: Investigators reviewed the health service's data base for diagnoses of conjunctivitis. Viral and bacterial cultures were obtained from ill students. Bile-soluble isolates that were susceptible to ethylhydrocupreine (optochin) and therefore were presumed to be pneumococci underwent serotyping, capsular staining, pulsed-field gel electrophoresis, a DNA probe, and multilocus sequence typing. A cohort study of risk factors was conducted with the use of the Internet. Control measures included distribution of alcohol-based hand gel and messages about prevention. RESULTS: Among 5060 students, 698 (13.8 percent) received a diagnosis of conjunctivitis from January 1, 2002, through April 12, 2002, including 22 percent of first-year students. Presumed pneumococci were isolated from 43.3 percent of conjunctival swabs (110 of 254); viral cultures performed on 85 specimens were negative. DNA probes and multilocus sequence typing confirmed that the organisms were pneumococci, although the bacteria did not have the characteristic capsule. On pulsed-field gel electrophoresis, strains were found to be identical to pneumococci that caused outbreaks of conjunctivitis in other parts of the country in 1980. Analysis of survey data from 1832 students indicated that close contact with a student with conjunctivitis, wearing contact lenses, membership on a sports team, and attending parties at or living in a fraternity or sorority house were associated with conjunctivitis. The rate of diagnosis of conjunctivitis declined after the implementation of control measures and after spring break. CONCLUSIONS: This large outbreak of conjunctivitis on a college campus was caused by an atypical, unencapsulated strain of S. pneumoniae that was identical to strains that had caused outbreaks two decades earlier. PMID- 12646669 TI - Images in clinical medicine. Appearance of intraocular silicone oil on computed tomography. PMID- 12646670 TI - Preventing complications of central venous catheterization. PMID- 12646671 TI - The chromogranin-secretogranin family. PMID- 12646672 TI - Case records of the Massachusetts General Medicine. Weekly clinicopathological exercises. Case 9-2003. An 18-year-old man with back and leg pain and a nondiagnostic biopsy specimen. PMID- 12646673 TI - Defective melanocortin 4 receptors in hyperphagia and morbid obesity. PMID- 12646674 TI - CD40 ligand--assessing risk instead of damage? PMID- 12646675 TI - Race and genomics. PMID- 12646676 TI - The importance of race and ethnic background in biomedical research and clinical practice. PMID- 12646677 TI - Risk of myocardial infarction and polymorphisms in candidate genes. PMID- 12646678 TI - Surgical treatment of esophageal cancer. PMID- 12646679 TI - Mass treatment of filariasis in New Guinea. PMID- 12646680 TI - High-frequency ventilation. PMID- 12646681 TI - Typhoid fever. PMID- 12646682 TI - Views of practicing physicians and the public on medical errors. PMID- 12646683 TI - Low-dose heparin for severe sepsis. PMID- 12646684 TI - PR interval and the response to enzyme-replacement therapy for Fabry's disease. PMID- 12646685 TI - Bisphosphonates and ocular inflammation. PMID- 12646686 TI - Site-specific mutagenesis in a homogeneous polyglutamine tract: application to spinocerebellar ataxin-3. AB - In recent years, nine neurodegenerative diseases have been found to be caused by the expansion of a CAG-triplet repeat in the coding region of the respective genes, resulting in lengthening of an otherwise harmless polyglutamine tract in the gene products. To facilitate structural studies of these disease mechanisms, a general protocol is described that allows site-specific mutations to be introduced into the polyglutamine tract. Based on 'cassette mutagenesis', this protocol involves engineering unique restriction sites into the flanking regions of the CAG repeat and subsequently replacing the wild-type CAG repeat with a double-stranded synthetic DNA fragment containing the desired mutations. This method was applied to the spinocerebellar ataxin-3 protein, such that the wild type amino acid sequence -Q(3)KQ(22)- was replaced by a -Q(9)CQ(9)- sequence. In this case, the incorporated cysteine residue can be exploited for various chemical modifications, lending the host glutamine repeat to many structural and biophysical techniques for the resolution of a specific residue. The method reported here bypasses many problems that can arise from PCR-based mutagenesis methods. PMID- 12646687 TI - Buried water molecules contribute to the conformational stability of a protein. AB - This study sought to attain a better understanding of the contribution of buried water molecules to protein stability. The 3SS human lysozyme lacks one disulfide bond between Cys77 and Cys95 and is significantly destabilized compared with the wild-type human lysozyme (4SS). We examined the structure and stability of the I59A-3SS mutant human lysozyme, in which a cavity is created at the mutation site. The crystal structure of I59A-3SS indicated that there were ordered new water molecules in the cavity created. The stability of I59A-3SS is 5.5 kJ/mol less than that of 3SS. The decreased stability of I59A-3SS (5.5 kJ/mol) is similar to that of Ile to Ala mutants with newly introduced water molecules in other globular proteins (6.3 +/- 2.1 kJ/mol), but is less than that of Ile/Leu to Ala mutants with empty cavities (13.7 +/- 3.1 kJ/mol). This indicates that water molecules partially compensate for the destabilization by decreasing hydrophobic and van der Waals interactions. These results provide further evidence that buried water molecules contribute to protein stability. PMID- 12646688 TI - HLA-A3 supermotif defined by quantitative structure-activity relationship analysis. AB - Activation of a cytotoxic T cell requires specific binding of antigenic peptides to major histocompatibility complex (MHC) molecules. This paper reports a study of peptides binding to members of the HLA-A3 superfamily using a recently developed 2D-QSAR method, called the additive method. Four alleles with high phenotype frequency were included in the study: A*0301, A*1101, A*3101 and A*6801. The influence of each of the 20 amino acids at each position of the peptide on binding was studied. A refined A3 supertype motif was defined in the study. PMID- 12646690 TI - Evolutionary engineering of a beta-Lactamase activity on a D-Ala D-Ala transpeptidase fold. AB - The beta-Lactamase hydrolytic activity has arisen several times from DD transpeptidases. We have been able to replicate the evolutionary process of beta Lactamase activity emergence on a PBP2X DD-transpeptidase. Some of the most interesting changes, like modifying the catalytic properties of an enzyme, may require several mutations in concert; therefore it is essential to explore efficiently sequence space by generating the right diversity. We designed a biased combinatorial library in which biochemical and structural information were incorporated by site directed mutagenesis on relevant residues and then subjected to random mutagenesis to allow for mutations in unforeseen positions. We isolated mutants from this library conferring 10-fold higher cefotaxime resistance levels than the background wild-type through mutations exclusively in the coding sequence. We demonstrate that only three substitutions in the DD-transpeptidase active site, two produced by the directed and one by the random mutagenesis, are sufficient to acquire this activity. The purified product of one mutant (MutE) had a 10(5)-fold increase in cefotaxime deacylation rate allowing it to hydrolyze beta-Lactams yet it has apparently conserved DD-peptidase activity. This work is the first to show a possible evolutionary intermediate between a beta-Lactamase and a DD-transpeptidase necessary for the development of antibiotic resistance. PMID- 12646689 TI - Protein engineering to improve the thermostability of glucoamylase from Aspergillus awamori based on molecular dynamics simulations. AB - Twelve mutations were constructed to improve the thermostability of glucoamylase from Aspergillus awamori based on the results of molecular dynamics simulations. The thermal unfolding of the catalytic domain followed a putative hierarchical behavior. In addition, the unfolding of the 13 alpha-helices obeyed the random ordered mechanism, in which the alpha-helices 8, 1 and 11 unfolded more rapidly than the others. The catalytic center was well protected by the (alpha/alpha)(6) barrel at simulation temperatures up to 600 K, whereas the catalytic base, E400, migrated from its original interior pocket to the surface of the catalytic domain by surmounting the hydrophobic barrier provided by alpha-helices 12 and 13 at 800 K. The disulfide bonds engineered to 'lock' the alpha-helix 11 on the surface of the catalytic domain dramatically increased the thermostability. Substituting G396 and G407 with Ala residues slightly increased the thermostability, whereas their specific activity and catalytic efficiency were reduced. This indicates that the introduced residues with higher hydrophobicity were favorable in the loop between alpha-helices 12 and 13, whereas they partially destroyed the hydrogen bond and salt linkage network in the catalytic center. Alpha-helices 12 and 13 can be stabilized by introducing residues with higher hydrophobicity, except for the H391M mutation. PMID- 12646691 TI - Further improvement of broad specificity hapten recognition with protein engineering. AB - Sulfa-antibiotics (sulfonamides) are widely used in veterinary medicine. Meat and milk from treated animals can be contaminated with sulfa residues. Current sulfonamide assays are unfit for screening of food, because they are either too laborious, insensitive or specific for a few sulfa compounds only. An immunoassay for detection of all sulfas in a single reaction would be useful for screening. Previously we have improved the broad specificity sulfa binding of antibody 27G3 with random mutagenesis and phage display. In order to improve the properties of this antibody further, mutants from the previous study were recombined and more mutations introduced. These new libraries were enriched with phage display and several different mutant antibodies were isolated. The cross-reaction profile of the best mutant was better than that of the wild-type antibody and the mutants of the previous study: it was capable of binding 10 of the tested 13 sulfonamides within a narrow concentration range and also bound the rest of the sulfas 5- to 11-fold better than the mutants of the previous study. PMID- 12646692 TI - Single-chain Fv multimers of the anti-neuraminidase antibody NC10: the residue at position 15 in the V(L) domain of the scFv-0 (V(L)-V(H)) molecule is primarily responsible for formation of a tetramer-trimer equilibrium. AB - Single-chain variable fragment of the murine monoclonal antibody NC10 specific to influenza virus N9 neuraminidase, joined directly in the V(L) to V(H) orientation (scFv-0), forms an equilibrium mixture of tetramer and trimer with the tetramer as the preferred multimeric species. In contrast, the V(H)-V(L) isomer was previously shown to exist exclusively as a trimer. Computer-generated trimeric and tetrameric scFv models, based on the refined crystal structure for NC10 Fv domain, were constructed and used to evaluate factors influencing the transition between V(L)-V(H) trimer and tetramer. These model structures indicated that steric restrictions between loops spanning amino acid residues L55-L59 and L13 L17 from the two adjacent V(L) domains within the V(L)-V(H) trimer were responsible for four scFv-0 molecules assembling to form a tetramer. In particular, leucine at position L15 and glutamate at position L57 appeared to interfere significantly with each other. To minimize this steric interference, the site-directed mutagenesis technique was used to construct several NC10 scFv-0 clones with mutations at these positions. Size-exclusion chromatographic analyses revealed that several of these mutations resulted in the production of NC10 scFv 0 proteins with significantly altered tetramer-trimer equilibrium ratios. In particular, introduction of a polar residue, such as asparagine or threonine, at position L15 generated a highly stable NC10 scFv-0 trimer. PMID- 12646693 TI - Translated products of tandem microgene repeats exhibit diverse properties also seen in natural proteins. AB - Repetitiousness is often observed in the primary and tertiary structures of proteins. We are intrigued by the potential role played by periodicity in the evolution of proteins and have created artificial repetitious proteins from repeats of short DNA sequences (microgenes). In this paper we characterize the physicochemical properties of six such artificially created proteins, which are the translated products of repeats of three microgenes. Three of the six proteins contain beta-sheet-like structures and are rather hydrophobic in nature. These proteins form macroscopic membranous structures in the presence of monovalent cationic ions, suggesting they have the capacity to promote strong intermolecular interactions. Of the other three proteins, one is comprised of alpha-helices and two have disordered structures. Small angle X-ray scattering analysis indicates that the artificial proteins do not fold as tightly as natural proteins, but are more compact than if completely denatured. One alpha-helical protein whose microgene unit was designed from coiled coil proteins was crystallized, demonstrating that repetitious artificial proteins can undergo transition to a more ordered state under appropriate conditions. Application of this approach to the development of a novel protein engineering system is discussed. PMID- 12646694 TI - Use of phage display to probe the evolution of binding specificity and affinity in integrins. AB - The specific binding of RGD-containing proteins to integrin is a function of both the conformation of and the local sequence surrounding the RGD motif. To study the effect of these factors on integrin binding affinity and specificity, we obtained RGD-containing ligands specific for different integrins presented on the same protein scaffold. The beta-turn region between two anti-parallel beta strands on the loop I of tendamistat, an inhibitor of alpha-amylase, was extended by two residues and randomized in a phagemid library. This library and two subsequently constructed RGD-containing loop I libraries were biopanned with purified integrins alphaIIbbeta3, alphaVbeta3 and alphaVbeta5 individually. The sequence analysis of selected tendamistat variants and characterization by phage ELISA revealed that phage adhesion is mediated exclusively by an RGD motif located at only two out of four possible positions on loop I. Further, sequences flanking the RGD motif were specific for different integrin targets. Interestingly, selected tendamistat variants mimic natural integrin ligands, both in sequence similarity and in integrin binding specificity, indicating that various ligand specificity patterns can be generated by driving towards maximum affinity in the integrin-ligand complexes. PMID- 12646695 TI - Baculovirus surface display of Theileria parva p67 antigen preserves the conformation of sporozoite-neutralizing epitopes. AB - Theileria parva is an intracellular protozoan parasite that causes East Coast fever, a severe lymphoproliferative disease in cattle. Previous attempts to produce recombinant sporozoite surface antigen (p67) in bacterial or insect cells for vaccine purposes have not resulted in a correctly folded protein. Here, we report the expression of N- and C-terminal domains of p67 fused to the baculovirus envelope glycoprotein GP64 by cloning the appropriate p67 cDNA segments between the signal sequence and the major portion of GP64. To further advance the generation of such recombinants, existing surface display techniques were combined with bacmid technology. Chimeric proteins were present on the surface of budded viruses as judged by immunogold labelling and were exposed on the surface of insect cells, as concluded from immunofluorescence studies of infected, non-fixed insect cells. In non-denaturing dot blot experiments, a strong reaction was obtained between monoclonal TpM12 and baculovirus particles displaying the p67N-GP64 chimeric protein. This antibody, raised against native p67, also specifically recognized the surface of recombinant-infected cells. Apparently, a more native conformation was achieved than when p67 was expressed in E.coli or in conventional baculovirus expression systems. The baculovirus surface expression system, therefore, provides an improved way of expressing this T.parva sporozoite surface protein. PMID- 12646698 TI - Property gamma factors and the Hochschild cohomology problem. AB - The main result of this article is that the kth continuous Hochschild cohomology groups H(k)(M,M ) and H(k)(M, B(H)) of a von Neumann factor subset, dbl equals B(H) of type II(1) with property Gamma are 0 for all positive integers k. The method of proof involves the construction of hyperfinite subfactors with special properties and a new inequality of Grothendieck type for multilinear maps. We prove joint continuity in the parallel. parallel (2) norm of separately ultraweakly continuous multilinear maps and combine these results to reduce to the case of completely bounded cohomology, which is already solved. PMID- 12646696 TI - Designing isoform-specific peptide disruptors of protein kinase A localization. AB - A kinase-anchoring proteins (AKAPs) coordinate cAMP-mediated signaling by binding and localizing cAMP-dependent protein kinase (PKA), using an amphipathic helical docking motif. Peptide disruptors of PKA localization that mimic this helix have been used successfully to assess the involvement of PKA in specific signaling pathways. However, these peptides were developed as disruptors for the type II regulatory subunit (RII) even though both RI and RII isoforms can bind to AKAPs and have discrete functions. To evaluate the effects of each localized isoform, we designed peptides that specifically bind to either RI or RII. Using a peptide array, we have defined the minimal binding sequence of dual specific-AKAP 2 (d AKAP2), which binds tightly to both RI and RII. Side-chain requirements for affinity and isoform specificity were evaluated by using a peptide substitution array where each position along the A kinase binding domain of d-AKAP2 was substituted by the other 19 l-amino acids. This array comprises 513 single-site substitution analogs of the d-AKAP2 sequence. Peptides containing single and multiple mutations were evaluated in a quantitative fluorescence binding assay and a cell-based colocalization assay. This strategy has allowed us to design peptides with high affinity (K(D) = 1-2 nM) and high specificity for RIalpha versus RIIalpha. These isoform-specific peptides will be invaluable tools to evaluate functional differences between localized RI and RII PKA and are RIalpha specific disruptors. This array-based analysis also provides a foundation for biophysical analysis of this docking motif. PMID- 12646697 TI - Amino acid variant in the kinase binding domain of dual-specific A kinase anchoring protein 2: a disease susceptibility polymorphism. AB - The focus of human genetics in recent years has shifted toward identifying genes that are involved in the development of common diseases such as cancer, diabetes, cardiovascular diseases, and Alzheimer's disease. Because many complex diseases are late-onset, the frequencies of disease susceptibility alleles are expected to decrease in the healthy elderly individuals of the population at large because of their contribution to disease morbidity andor mortality. To test this assumption, we compared allele frequencies of 6,500 single-nucleotide polymorphisms (SNPs) located in approximately 5,000 genes between DNA pools of age-stratified healthy, European-American individuals. A SNP that results in an amino acid change from Ile to Val in the dual-specific A kinase-anchoring protein 2 (d-AKAP2) gene, showed the strongest correlation with age. Subsequent analysis of an independent sample indicated that the Val variant was associated with a statistically significant decrease in the length of the electrocardiogram PR interval. The IleVal SNP is located in the A-kinase-binding domain. An in vitro binding assay revealed that the Ile variant bound approximately 3-fold weaker to the protein kinase A (PKA)-RIalpha isoform than the Val variant. This decreased affinity resulted in alterations in the subcellular distribution of the recombinantly expressed PKA-RIalpha isoform. Our study suggests that alterations in PKA-RIalpha subcellular localization caused by variation in d-AKAP2 may have a negative health prognosis in the aging population, which may be related to cardiac dysfunction. Age-stratified samples appear to be useful for screening SNPs to identify functional gene variants that have an impact on health. PMID- 12646699 TI - Posterior parietal cortex and the filtering of distractors. AB - Neural systems for visual processing can focus attention on behaviorally relevant objects, filtering out competing distractors. Neurophysiological studies in animals and brain imaging studies in humans suggest that such filtering depends on top-down inputs to extrastriate visual areas, originating in structures important for attentional control. To test whether the posterior parietal cortex may be a necessary source of signals that filter distractors, we measured the ability of a patient with bilateral parietal lesions to discriminate the features of a target surrounded by distractors of variable contrast. In the presence of distractors, the patient was impaired at discriminating both grating orientation and faces, and the magnitude of the impairment increased with distractor salience. These attentional deficits are remarkably similar to those caused by damage to monkey extrastriate regions V4 andor TEO, which are thought to be recipients of top-down attentional feedback. In contrast to the effects of V4 and TEO lesions, however, the parietal lesions impaired performance even with widely spaced targets and distractors, a finding consistent with the projections of parietal cortex to visual processing areas covering a wide range of receptive field sizes and eccentricities. PMID- 12646700 TI - Transforming growth factor beta 1 inhibits expression of NKp30 and NKG2D receptors: consequences for the NK-mediated killing of dendritic cells. AB - The surface density of the triggering receptors responsible for the natural killer (NK)-mediated cytotoxicity is crucial for the ability of NK cells to kill susceptible target cells. In this study, we show that transforming growth factor beta1 (TGFbeta1) down-regulates the surface expression of NKp30 and in part of NKG2D but not that of other triggering receptors such as NKp46. The TGFbeta1 mediated inhibition of NKp30 surface expression reflects gene regulation at the transcriptional level. NKp30 has been shown to represent the major receptor involved in the NK-mediated killing of dendritic cells. Accordingly, the TGFbeta1 dependent down-regulation of NKp30 expression profoundly inhibited the NK mediated killing of dendritic cells. On the contrary, killing of different NK susceptible tumor cell lines was variably affected, reflecting the differential usage of NKp30 and/or NKG2D in the lysis of such tumors. Our present data suggest a possible mechanism by which TGFbeta1-producing dendritic cells may acquire resistance to the NK-mediated attack. PMID- 12646701 TI - Mineralized tissue and vertebrate evolution: the secretory calcium-binding phosphoprotein gene cluster. AB - Gene duplication creates evolutionary novelties by using older tools in new ways. We have identified evidence that the genes for enamel matrix proteins (EMPs), milk caseins, and salivary proteins comprise a family descended from a common ancestor by tandem gene duplication. These genes remain linked, except for one EMP gene, amelogenin. These genes show common structural features and are expressed in ontogenetically similar tissues. Many of these genes encode secretory Ca-binding phosphoproteins, which regulate the Ca-phosphate concentration of the extracellular environment. By exploiting this fundamental property, these genes have subsequently diversified to serve specialized adaptive functions. Casein makes milk supersaturated with Ca-phosphate, which was critical to the successive mammalian divergence. The innovation of enamel led to mineralized feeding apparatus, which enabled active predation of early vertebrates. The EMP genes comprise a subfamily not identified previously. A set of genes for dentine and bone extracellular matrix proteins constitutes an additional cluster distal to the EMP gene cluster, with similar structural features to EMP genes. The duplication and diversification of the primordial genes for enameldentinebone extracellular matrix may have been important in core vertebrate feeding adaptations, the mineralized skeleton, the evolution of saliva, and, eventually, lactation. The order of duplication events may help delineate early events in mineralized skeletal formation, which is a major characteristic of vertebrates. PMID- 12646702 TI - The protein-folding speed limit: intrachain diffusion times set by electron transfer rates in denatured Ru(NH3)5(His-33)-Zn-cytochrome c. AB - The kinetics of electron transfer from the triplet-excited Zn-porphyrin to a Ru(NH(3))(5)(His-33)(3+) complex have been measured in Zn-substituted ruthenium modified cytochrome c under denaturing conditions. In the folded protein, the electron-tunneling rate constant is 7.5 x 10(5) s(-1). As the protein is denatured with guanidine hydrochloride, a faster adiabatic electron-transfer reaction appears (4.0 x 10(6) s(-1), [guanidine hydrochloride] = 5.4 M) that is limited by the rate of intrachain diffusion to bring the Zn-porphyrin and Ru complex into contact. The 250-ns contact time for formation of a 15-residue loop in denatured cytochrome c is in accord with a statistical model developed by Camacho and Thirumalai [Camacho, C. J. & Thirumalai, D. (1995) Proc. Natl. Acad. Sci. USA 92, 1277-1281] that predicts that the most probable transient loops formed in denatured proteins are comprised of 10 amino acids. Extrapolation of the cytochrome c contact time to a 10-residue loop sets the folding speed limit at approximately 10(7) s(-1). PMID- 12646703 TI - A macrocyclic zinc(II) fluorophore as a detector of apoptosis. AB - Our originally designed dansylamidoethylcyclen 4 as a biomimetic Zn(2+)-selective fluorophore has been demonstrated to be a good detector of the apoptosis (induced by an anticancer agent, etoposide, and H(2)O(2)) in cancer cells such as HeLa and HL60 cells. The macrocyclic Zn(2+) ligand 4 (mostly as a deprotonated form) is cell-permeable to show weak fluorescence (emission at 550 nm), which forms a strong fluorescent 1:1 Zn(2+) complex 5 (emission at 530 nm) when Zn(2+) is incorporated into the cells by a zinc(II) ionophore pyrithione. Thus formed, Zn(2+) complex 5 is cell-impermeable and remains intact over a few hours. When apoptosis in HeLa or HL60 cells is artificially induced, 4 selectively and strongly stains apoptotic cells only at early stages, which was verified by using the conventional apoptosis detection probe annexin V-Cy3. Detection of the apoptotic cells by 4 was perhaps due to significantly increased free Zn(2+) flux at early stages of apoptosis. Apoptotic detection by 4 has been compared with a presently available Zn(2+) fluorophore, Zinquin 1. We present that 4 has advantages in detection of apoptosis over annexin V-Cy3 and Zinquin 1. PMID- 12646704 TI - Inducible protein knockout reveals temporal requirement of CaMKII reactivation for memory consolidation in the brain. AB - By integrating convergent protein engineering and rational inhibitor design, we have developed an in vivo conditional protein knockout andor manipulation technology. This method is based on the creation of a specific interaction interface between a modified protein domain and sensitized inhibitors. By introducing this system into genetically modified mice, we can readily manipulate the activity of a targeted protein, such as alpha-Ca(2+)calmodulin-dependent protein kinase II (alphaCAMKII), on the time scale of minutes in specific brain subregions of freely behaving mice. With this inducible and region-specific protein knockout technique, we analyzed the temporal stages of memory consolidation process and revealed the first postlearning week as the critical time window during which a precise level of CaMKII reactivation is essential for the consolidation of long-term memories in the brain. PMID- 12646705 TI - A soluble protein is immobile in dormant spores of Bacillus subtilis but is mobile in germinated spores: implications for spore dormancy. AB - Fluorescence redistribution after photobleaching has been used to show that a cytoplasmic GFP fusion is immobile in dormant spores of Bacillus subtilis but becomes freely mobile in germinated spores in which cytoplasmic water content has increased approximately 2-fold. The GFP immobility in dormant spores is not due to the high levels of dipicolinic acid in the spore cytoplasm, because GFP was also immobile in germinated cwlD spores that had excreted their dipicolinic acid but where cytoplasmic water content had only increased to a level similar to that in dormant spores of several other Bacillus species. The immobility of a normally mobile protein in dormant wild-type spores and germinated cwlD spores is consistent with the lack of metabolism and enzymatic activity in these spores and suggests that protein immobility, presumably due to low water content, is a major reason for the metabolic dormancy of spores of Bacillus species. PMID- 12646706 TI - Gigantic cannibals driving a whole-lake trophic cascade. AB - Trophic cascades have been a central paradigm in explaining the structure of ecological communities but have been demonstrated mainly through comparative studies or experimental manipulations. In contrast, evidence for shifts in trophic cascades caused by intrinsically driven population dynamics is meager. By using empirical data of a cannibalistic fish population covering a 10-year period and a size-structured population model, we show the occurrence of a dynamic trophic cascade in a lake ecosystem, in which the community over time alternates between two different configurations. The intrinsically driven change in the size structure of the fish population from a dominance of stunted individuals to a dominance of gigantic cannibals among adult individuals is the driving force behind distinct abundance switches observed in zooplankton and phytoplankton. The presence of the phase with gigantic cannibals depends critically on the energy they extract from their victims, allowing strong reproduction for a number of years. PMID- 12646707 TI - The CsrR/CsrS two-component system of group A Streptococcus responds to environmental Mg2+. AB - Group A streptococci control expression of key virulence determinants via the two component sensorregulator system CsrRCsrS. The membrane-bound sensor CsrS is thought to respond to previously unknown environmental signal(s) by controlling phosphorylation of its cognate regulator component CsrR. Phosphorylation of CsrR increases its affinity for binding to the promoter regions of Csr-regulated genes to repress transcription. Here we show that environmental Mg(2+) concentration is a potent and specific stimulus for CsrRCsrS-mediated regulation. We studied the effect of divalent cations on expression of the Csr-regulated hyaluronic acid capsule genes (hasABC) by measuring chloramphenicol acetyltransferase (CAT) activity in a reporter strain of group A Streptococcus carrying a has operon promoter-cat fusion. Addition of Mg(2+), but not of Ca(2+), Mn(2+), or Zn(2+), repressed capsule gene expression by up to 80% in a dose-dependent fashion. The decrease in capsule gene transcription was associated with a marked reduction in cell-associated capsular polysaccharide. RNA hybridization analysis demonstrated reduced expression of the Csr-regulated hasABC operon, streptokinase (ska), and streptolysin S (sagA) during growth in the presence of 15 mM Mg(2+) for the wild type strain 003CAT but not for an isogenic csrS mutant. We propose that Mg(2+) binds to CsrS to induce phosphorylation of CsrR and subsequent repression of virulence gene expression. The low concentration of Mg(2+) in extracellular body fluids predicts that the CsrRCsrS system is maintained in the inactive state during infection, thereby allowing maximal expression of critical virulence determinants in the human host. PMID- 12646708 TI - The influence of the synergistic anion on iron chelation by ferric binding protein, a bacterial transferrin. AB - Although the presence of an exogenous anion is a requirement for tight Fe(3+) binding by the bacterial (Neisseria) transferrin nFbp, the identity of the exogenous anion is not specific in vitro. nFbp was reconstituted as a stable iron containing protein by using a number of different exogenous anions [arsenate, citrate, nitrilotriacetate, pyrophosphate, and oxalate (symbolized by X)] in addition to phosphate, predominantly present in the recombinant form of the protein. Spectroscopic characterization of the Fe(3+)anion interaction in the reconstituted protein was accomplished by UV-visible and EPR spectroscopies. The affinity of the protein for Fe(3+) is anion dependent, as evidenced by the effective Fe(3+) binding constants (K'(eff)) observed, which range from 1 x 10(17) M(-1) to 4 x 10(18) M(-1) at pH 6.5 and 20 degrees C. The redox potentials for Fe(3+)nFbpXFe(2+)nFbpX reduction are also found to depend on the identity of the synergistic anion required for Fe(3+) sequestration. Facile exchange of exogenous anions (Fe(3+)nFbpX + X' --> Fe(3+)nFbpX' + X) is established and provides a pathway for environmental modulation of the iron chelation and redox characteristics of nFbp. The affinity of the iron loaded protein for exogenous anion binding at pH 6.5 was found to decrease in the order phosphate > arsenate approximately pyrophosphate > nitrilotriacetate > citrate approximately oxalate carbonate. Anion influence on the iron primary coordination sphere through iron binding and redox potential modulation may have in vivo application as a mechanism for periplasmic control of iron delivery to the cytosol. PMID- 12646709 TI - Dietary zinc modulates gene expression in murine thymus: results from a comprehensive differential display screening. AB - Differential mRNA display was used to comprehensively screen the murine thymic transcriptome for genes modulated in vivo by dietary zinc. A moderate feeding protocol rendered young adult, outbred mice zinc-deficient and zinc-supplemented without alterations in feeding behavior or growth. However, these levels of deficiency and supplementation altered specific mRNA abundances in a manner detectable by differential display. In total, 240 primer-pair combinations were used to generate >48,000 interpretable cDNA bands derived from thymic total RNA, of which only 265 or 0.55% were identified as zinc-modulated under these moderate dietary conditions. The most strongly zinc-modulated cDNAs identified by display were reamplified and sequenced. No cDNAs encoding zinc-metalloenzymes or zinc finger transcription factors were identified as zinc-modulated in this global screening. Those zinc-regulated genes independently confirmed by quantitative PCR included: heat shock proteins 40 and 60; heat shock cognate 70; histocompatibility 2, class II antigen A, alpha; and the T cell cytokine receptor. In addition, a variety of transcription- and translation-related factors (such as ribosomal proteins L3, L5, and L28; nuclear matrix protein 84; matrin cyclophilin; the H3 histone family 3A protein; beta(2) microglobulin; and a cleavage and polyadenylation factor) were identified as zinc-modulated. These profiling data show that differential expression of genes in the thymus in response to the dietary zinc supply precedes many of the phenotypic effects on thymic function associated with severe zinc restriction or supplementation. Several genes involved in T cell development were identified as regulated by zinc and will be targets to evaluate the effects of zinc on immune function. PMID- 12646711 TI - Database for renal collecting duct regulatory and transporter proteins. AB - The mammalian kidney collecting duct plays an important role in the fine regulation of Na, K, water, and acid-base balance. Functional genomic and proteomic studies of the kidney offer new opportunities in the understanding of renal physiology and pathophysiology, and the collecting duct is an appropriate target tissue because of the relative simplicity of its cells and the ease of isolating or culturing large numbers of collecting duct cells. Study of the collecting duct includes assessment of gene expression and protein regulation and abundance. For example, DNA and protein microarrays can be used to quantitate gene expression and protein regulation and abundance under varying physiological conditions. An Internet-accessible database has been devised for major collecting duct proteins involved in transport and regulation of cellular processes. The individual proteins included in this database are those culled from literature searches and from previously published studies involving cDNA arrays and serial analysis of gene expression (SAGE). Design of microarray targets for the study of kidney collecting duct tissues is facilitated by the database, which includes links to curated base pair and amino acid sequence data, relevant literature, and related databases. Use of the database is illustrated by a search for water channel proteins, aquaporins, and by a subsequent search for vasopressin receptors. Links are shown to the literature and to sequence data for human, rat, and mouse, as well as to relevant web-based resources. Extension of the database is dynamic and is done through a maintenance interface. This permits creation of new categories, updating of existing entries, and addition of new ones. PMID- 12646710 TI - Phosphatidylinositol 4-kinase type IIalpha is responsible for the phosphatidylinositol 4-kinase activity associated with synaptic vesicles. AB - Phosphorylation of inositol phospholipids plays a key role in cellular regulation via the generation of intracellular second messengers. In addition, it represents a mechanism to regulate interactions of the lipid bilayer with proteins and protein scaffolds involved in vesicle budding, cytoskeletal organization, and signaling. Generation of phosphatidylinositol 4-phosphate [PI(4)P] from phosphatidylinositol (PI) is an important step in this metabolic pathway because PI(4)P is a precursor of other important phosphoinositides and has protein binding properties of its own. We report here that a PI 4-kinase (PI4K) activity previously reported on synaptic vesicles is accounted for by the alpha isoform of the recently characterized type II PI4K (PI4KII) family. PI4KIIalpha, which also accounts for the bulk of PI4K activity in brain extracts, is concentrated at synapses and in the region of the Golgi complex in neuronal perikarya. Our results provide new evidence for the occurrence of a cycle of phosphoinositide synthesis and hydrolysis nested within the exo-endocytic cycle of synaptic vesicles and point to PI4KIIalpha as a critical player in this cycle. PMID- 12646713 TI - Different relationship between anthropometric markers and umbilical cord plasma leptin in Asian and Caucasian neonates. AB - The leptin to fat ratio early in life could contribute to fixing the set point of leptin feedback at the hypothalamic level. Subjects from Asian and Caucasian ethnicities differ in body composition. We tested the hypothesis that anthropometric markers and their relationship to umbilical cord leptin, cortisol and cortisone, DHEAs and oestriol differed between Caucasians and Asians at birth. Birthweight, length, arm, calf and abdominal circumferences, scapular, triceps, quadriceps and abdominal skinfolds were measured in 180 healthy, full term newborns of Asian and Caucasian ethnicities. Leptin and steroid hormone concentrations were determined in umbilical cord plasma. There was a significant difference in the slope of the regression between leptin and birthweight (p = 0.03) and calf circumference (p = 0.05) between male Caucasian and Asian neonates. In contrast, in female neonates, there was no significant difference (p = 0.099 and p = 0.07 for birthweight and calf circumference, respectively). In addition, while the slopes of the regression plots were not affected by gender in Asian newborns, there was a significant difference between male and female Caucasian newborns (p = 0.006 and p = 0.002 for birthweight and calf circumference, respectively). There was no significant correlation between cord leptin concentrations or anthropometric markers and steroid hormone concentrations. In conclusion, gender and ethnic differences in the relationship between leptin and anthropometric markers are detectable at birth between Asians and Caucasians, two ethnic groups that have been demonstrated to have different body compositions later in life. This may represent the first clinical evidence of a difference in leptin regulation between these two ethnic groups. PMID- 12646712 TI - Reassessment of the unbound concentrations of unconjugated bilirubin in relation to neurotoxicity in vitro. AB - Most studies of the cellular toxicity of unconjugated bilirubin (UCB) have been performed at concentrations of unbound UCB (BF) that exceed those in the plasma of neonates with bilirubin encephalopathy. We assessed whether UCB could be toxic to neurons and astrocytes at clinically relevant BF values (7 d. We defined episodes of sepsis and sepsis-like illness as acute clinical deteriorations that prompted physicians to obtain blood cultures and start antibiotics. A predictive statistical model yielding an HRC index was developed on a derivation cohort of 316 neonates in the University of Virginia NICU and then applied to the validation cohort of 317 neonates in the Wake Forest University NICU. In the derivation cohort, there were 155 episodes of sepsis and sepsis-like illness in 101 infants, and in the validation cohort, there were 118 episodes in 93 infants. In the validation cohort, the HRC index 1) showed highly significant association with impending sepsis and sepsis-like illness (receiver operator characteristic area 0.75, p < 0.001) and 2) added significantly to the demographic information of birth weight, gestational age, and days of postnatal age in predicting sepsis and sepsis-like illness (p < 0.001). Continuous HRC monitoring is a generally valid and potentially useful noninvasive tool in the early diagnosis of neonatal sepsis and sepsis-like illness. PMID- 12646727 TI - Functional and molecular characterization of rat intestinal prolidase. AB - Genetic deficiency of prolidase can lead to severe problems in child development, including mental retardation. However, the exact pathogenesis of the disease is unclear. To understand the enzyme's physiologic functions, we studied the regulation of rat intestinal prolidase. The results indicated that 1) the activities of intestinal prolidase and its kinetic parameters (Km and Vmax) are site-dependent; 2) the jejunal prolidase activity was the most sensitive to the dietary restriction, and the duodenal and jejunal but not colonic kinetic parameters changed with dietary restriction; 3) the pH activity profile of jejunal prolidase at 24 h postfeeding was different from that at 48 h postfeeding, whereas the inhibition profiles of prolidase were qualitatively independent of dietary restriction; and 4) old-aged rats have lower prolidase activities in the small intestine. We also purified rat intestinal prolidase I to homogeneity. The characterization study indicated that the purified rat intestinal prolidase I is fairly similar to prolidase I from other species with a molecular weight of 116,000, which consisted of two monomers, 58,000 D each. The purified prolidase I has a Km value of 178 microM and a Vmax value of 601 micromol x min-1. mg protein-1. Screening of a rat intestinal cDNA library produced a 1.8-kb fragment that encodes the rat intestinal prolidase. This enzyme has 494 deduced amino acid sequence, which is 96% or 86% identical to mouse or human erythrocyte prolidase I. This represents the first report of a successful attempt to purify and clone an intestinal prolidase and of investigation to study prolidase regulation by diet. PMID- 12646728 TI - Characteristic acylcarnitine profiles in inherited defects of peroxisome biogenesis: a novel tool for screening diagnosis using tandem mass spectrometry. AB - Patients with inherited defects of peroxisomal metabolism, a class of diseases with marked clinical and genetic heterogeneity, show a characteristic phenotype in most cases with severe neurologic impairment, craniofacial abnormalities, and hepatic and kidney dysfunction. For the differential diagnosis of clinically suspected cases, a complex biochemical and genetic approach is required. Analysis of plasma very-long-chain fatty acids is a reliable screening method to detect most but not all peroxisomal disorders. To study the potential presence of abnormal acylcarnitine species in plasma and blood, we screened by tandem mass spectrometry a series of patients affected by a peroxisome biogenesis disorder (PBD) and compared the results with those obtained in patients with isolated peroxisomal defects (e.g. D-bifunctional protein deficiency, X-linked adrenoleukodystrophy) and mitochondrial long-chain fatty acid oxidation defects. The most relevant finding observed in plasma of patients with PBD was a significant increase of long-chain dicarboxylic C16- and C18-carnitine, i.e. hexadecanedioyl- and octadecanedioyl-carnitine, with high dicarboxylycarnitine/monocarboxylylcarnitine ratio. Elevation of very long-chain acylcarnitines C24- and C26-, i.e. lignoceroyl- and cerotoyl-carnitine, was detected in some PBDs and in D-bifunctional protein deficiency. Similar abnormalities were also found in neonatal screening blood spots. Detection of these compounds alone, in the absence of other shorter-chain acylcarnitines, is highly specific and characteristic of PBD, as confirmed by the differing profiles observed in patients with adrenoleukodystrophy and mitochondrial long-chain fatty acid oxidation defects. Our study adds a novel method to the diagnosis of PBD, which may also be of benefit for future neonatal mass screening programs based on acylcarnitine profiling. PMID- 12646729 TI - The basomedial hypothalamus modulates the ventilatory response to hypoxia in neonatal rats. AB - We sought to examine the role of the basomedial hypothalamus in the regulation of breathing in neonatal rats. Small basomedial hypothalamic lesions were produced by a lesioning agent, goldthioglucose, in 5-d-old male rat pups, and 2 d later, baseline ventilation and the ventilatory responses to hypoxia and hypercapnia were examined. When compared with vehicle-injected controls, goldthioglucose lesioned rat pups had a significantly slower respiratory rate and longer expiratory time at baseline. Lesioned rats displayed an impaired increase in breathing frequency in response to hypoxia, and augmented increases in tidal volume and respiratory drive (the ratio of tidal volume to inspiratory time) during hypoxia relative to controls. Hypercapnic responses were not affected. These data demonstrate that cells in a restricted area of the hypothalamus specifically influence the respiratory response to hypoxia. PMID- 12646730 TI - IUGR alters postnatal rat skeletal muscle peroxisome proliferator-activated receptor-gamma coactivator-1 gene expression in a fiber specific manner. AB - Uteroplacental insufficiency and subsequent intrauterine growth retardation (IUGR) increase the risk of insulin resistance in humans and rats. Aberrant skeletal muscle lipid metabolism contributes to the pathogenesis of insulin resistance. Peroxisome proliferator-activated receptor-gamma co-activator-1 (PGC 1) is a transcriptional co-activator that affects gene expression of key lipid metabolizing enzymes such as carnitine palmitoyl-transferase I (mCPTI). Because gene expression of lipid metabolizing enzymes is altered in IUGR postnatal skeletal muscle, and we hypothesized that PGC-1 expression would be similarly affected. To prove this hypothesis, bilateral uterine artery ligation and sham surgery were used to produce IUGR and control rats respectively. Western Blotting demonstrated that PGC-1 hind limb skeletal muscle protein levels were increased in perinatal and postnatal IUGR rats. Conventional RT-PCR demonstrated that PGC-1 mRNA levels were similarly increased in perinatal hind limb skeletal muscle and juvenile extensor digitorum longus (EDL), but were decreased in juvenile soleus. Because a gender specific trend was noted in PGC-1 mRNA levels, real time RT-PCR was used for further differentiation. Real time RT-PCR revealed that changes in postnatal skeletal muscle PGC-1 expression were more marked in male IUGR rats versus female IUGR rats. Down stream targets of PGC-1 followed a similar pattern of expression. We conclude that PGC-1 expression is altered in rat IUGR skeletal muscle and speculate that it contributes to the pathogenesis of insulin resistance in the IUGR rat. PMID- 12646732 TI - The effect of Escherichia coli endotoxin infusion on the ventilatory response to hypoxia in unanesthetized newborn piglets. AB - To determine the effects of endotoxemia on the neonatal ventilatory response to hypoxia, 17 chronically instrumented and unanesthetized newborn piglets (2 ng/ml (liposomes) and <150 ng/ml (adenovirus) per 5,000 initially planted BCEC]. Expression levels in BCEC were dependent on the concentration of applied liposomes. For gene transfer in HC, only the adenoviral transduction technique achieved a high production of vIL-10, whereas liposomal transfection led only to low vIL-10 secretion (4.8 microg/ml vs. 95 pg/ml per quarter of cornea). CONCLUSION: For transfection of corneal endothelial cells in culture, liposomes can be considered as a safe and useful alternative method of gene transfer avoiding side-effects of viral vectors. However, for transfection of organ-cultured HC, adenoviral vectors are superior to liposomal vehicles. PMID- 12646754 TI - Positron emission tomographic study of post-ischaemic-hypoxic amnesia. AB - BACKGROUND: Despite extensive research, it still remains controversial as to what the precise location of the critical lesions underlying amnesia actually is. The amnesic syndrome is believed to be heterogeneous and due to several distinct functional deficits. PATIENTS AND METHODS: Two patients, a 45-year-old woman and a 56-year-old man, with sudden cardiopulmonary arrest and successful resuscitation, were left with a clear amnesic syndrome as main neurological sequela. During their revalidation period, they underwent a positron emission tomographic (PET) examination, utilizing the (13)NH(3) bolus technique at rest and after intravenous acetazolamide administration. RESULTS: Both PET studies showed more or less similar features with a decrease in regional cerebral blood flow (rCBF) in the frontal, temporal and parietal lobes. In addition, the rCBF was increased in both thalami of the 45-year-old woman and in the striata of the 56-year-old man. Acetazolamide vasoreactivity was most lost in the frontal lobes. CONCLUSIONS: In the present PET study, we demonstrated that destruction of the inhibitory pathways to the thalamus and basal ganglia by ischaemic-hypoxic frontal lesions could be one of the mechanisms leading to amnesia. PMID- 12646755 TI - Effects of mitoxantrone on multiple sclerosis patients' lymphocyte subpopulations and production of immunoglobulin, TNF-alpha and IL-10. AB - We designed this longitudinal study to clarify the short- and long-term effects of mitoxantrone on the immune system in a subgroup of multiple sclerosis patients treated at our centre. After 14 days we found a highly significant sustained reduction of leucocytes, primarily affecting neutrophils and most lymphocyte subsets except for naive and activated T lymphocytes. The CD4/CD8 ratio and serum immmunoglobulin levels were not affected. Furthermore, whole blood-stimulated mononuclear cell IL-10 production showed a significant lower level 2 weeks treatment, whereas basal IL-10 as well as stimulated and basal TNF-alpha secretion showed no significant changes. Longitudinal data disclosed a persistent decrease of B lymphocytes, while secretion of immunoglobulins, IL-10, and TNF alpha was not altered in the follow-up. In conclusion, we confirmed a selective short-term effect of mitoxantrone therapy on most lymphocyte subpopulations, but not on immunoglobulines or the pro- and anti-inflammatory cytokines TNF-alpha and IL-10, which do not serve as possible response markers. PMID- 12646756 TI - Statistical parametric mapping of brain SPECT perfusion abnormalities in patients with Alzheimer's disease. AB - Brain perfusion in 20 patients with mild Alzheimer's disease (AD), 20 patients with moderate AD and 20 control subjects (matched for age, gender and education) were assessed by single photon emission computed tomography (SPECT) using technetium-99m hexamethylpropylene amine oxime ((99m)Tc-HMPAO). SPECT images were transformed to a standard size and shape for group comparisons by the voxel-based t test of the statistical parametric mapping techniques. Cerebral hypoperfusion in the left lower parietal area was found in mild AD patients. In moderate AD patients, significant cerebral hypoperfusion was located in bilateral posterior parietotemporal cortices, contiguous anterior occipital lobes, posterior cingulate gyri and, to a lesser extent, in frontal areas. PMID- 12646757 TI - Spinal MRI supporting myelopathic origin of early symptoms in unsuspected cobalamin deficiency. AB - We report two patients with subjectively progressive sensory symptoms and gait disturbance due to cobalamin deficiency, but only slight or absent abnormalities on neurological examination. In both patients, spinal MRI provided evidence for a myelopathic origin of the symptoms, disclosing characteristic T(2) hyperintense signal alterations confined to the posterior columns of the cervical and thoracic spinal cord. The patients illustrate the early clinical presentation of subacute combined degeneration (SCD) with a sensory neuropathy starting with acroparesthesia and Lhermitte's sign. Furthermore, the diagnostic value of spinal MRI for early diagnosis of SCD with characteristic findings is highlighted. PMID- 12646758 TI - Early antitoxin treatment in wound botulism results in better outcome. AB - Wound botulism in 7 heroin 'skin poppers' produced ophthalmoplegia and descending paralysis. Rapid recovery occurred in 2 who received the antitoxin within the fourth day of symptom onset. A poor outcome was seen in 4 who received the antitoxin after the eighth day of symptoms and 1 who did not receive the antitoxin. Early antitoxin administration is important in achieving a favorable outcome. PMID- 12646759 TI - Laryngeal stridor in multiple system atrophy. AB - The purpose of this study was to elucidate the clinical features of laryngeal stridor in 104 patients with multiple system atrophy (MSA) and to predict the hazard risk. Stridor was observed in 36 patients. It occurred in the first year of the disease in 10 cases, and 69% of the cases were diagnosed with stridor within the first 4 years. Dysphagia and hoarseness had a statistically higher frequency in the stridor group, and the onset period of these elements correlated with the onset of stridor. A follow-up study of survival probability was carried out in 83 patients. The median survival period in the stridor group (33 cases) and the non-stridor group (50 cases) was 8.0 and 9.0 years, respectively. Treatment for stridor decreased the relative risk from 2.998 to 0.147. Laryngeal stridor is a common and early clinical symptom in MSA. Early treatment for stridor is advisable to reduce mortality. PMID- 12646760 TI - Interleukin-2 production by peripheral blood mononuclear cells from patients with myasthenia gravis. AB - We examined interleukin-2 (IL-2) production by phytohemagglutinin (PHA) stimulated peripheral blood mononuclear cells (PBMs) from 75 untreated myasthenia gravis (MG) patients and 48 control patients. Patients with MG consisted of those with elevated PBM IL-2 production (>1,250 pg/ml; mean + 2SD of the controls) (n = 29, 39%) and those with normal PBM IL-2 production (<1,250 pg/ml) (n = 46, 61%). Significant characteristics of patients with elevated PBM IL-2 production included elevated serum levels of anti-acetylcholine receptor antibodies, severe generalized symptoms, thymic hyperplasia, and marked effects of thymectomy (p < 0.05). These findings suggest that elevated PBM IL-2 production can reflect functional abnormalities of T cells in some patients with MG, and that PBM IL-2 production should be considered as a candidate target of therapy. PMID- 12646761 TI - Extraocular blood flow and endothelin-1 plasma levels in patients with multiple sclerosis. AB - In order to evaluate whether plasma levels of the potent vasoconstrictor endothelin-1 (ET-1) are increased in patients with multiple sclerosis (MS) and whether these patients exhibit an ET-1-mediated vascular dysregulation, ET-1 plasma levels were measured in 30 patients with MS. Blood flow velocities in the ophthalmic artery, central retinal artery, central retinal vein, short lateral posterior ciliary artery, and short medial posterior ciliary artery were assessed in parallel. ET-1 plasma levels were significantly increased in MS patients when compared to sex- and age-matched healthy controls (2.0 +/- 0.4 pg/ml, range 1.1 2.8 vs. 1.5 +/- 0.2 pg/ml, range 0.9-2.0; p < 0.001). Moreover, the patients exhibited significant alterations of extraocular blood flow. The role of ET-1 in the inflammatory process remains to be clarified. PMID- 12646762 TI - Hypersomnia associated with bilateral posterior hypothalamic lesion. A polysomnographic case study. AB - We examined an obese 58-year-old patient with a bilateral posterior hypothalamic lesion of unknown etiology. A 24-hour polysomnography revealed a markedly increased total sleep time (17.6 h). During daytime, only 3 continuous wake phases occurred. REM periods occurred only between 5 p.m. and 6 a.m. We conclude from our results that, similar to the results from animal experiments, the posterior hypothalamus in humans plays a critical role in the maintenance of wakefulness. PMID- 12646763 TI - Comparative single-dose pharmacokinetics of clonazepam following intravenous, intramuscular and oral administration to healthy volunteers. AB - The objective was to assess the single-dose pharmacokinetics of clonazepam following i.m., p.o. and i.v. administration. In an open-label, three-way crossover study, 12 healthy volunteers were randomized to receive a single dose of 2 mg clonazepam either by the i.m., p.o. or i.v. route. Serial blood samples were collected up to 120 h after drug administration. Plasma concentrations of clonazepam were determined by electron-capture gas-liquid chromatography. The absorption rates of clonazepam after i.m. and p.o. administration of clonazepam were significantly different from each other, as reflected by the respective mean values of maximum plasma concentration (C(max) 11.0 vs. 14.9 ng.ml(-1)) and time to reach maximum concentration (t(max) 3.1 vs. 1.7 h). Secondary plasma peaks of clonazepam were observed in 9 volunteers after i.m. injection (C(max) 9.9 ng.ml( 1); t(max) 10.4 h). A comparison of the area under the plasma concentration-time curves (AUC) shows that the i.m. route is equivalent to the oral route (AUC(0- infinity ) 620 vs. 561 ng.h.ml(-1)). Clonazepam was almost completely absorbed after i.m. and p.o. administration, as shown by the mean absolute bioavailability of 93 and 90%, respectively. No significant differences existed between the elimination half-lives (i.v. 38.0 h; i.m. 43.6 h; p.o. 39.0 h). The average clearance and volume of distribution (V(Z)) were 55 ml.min(-1) and 180 liters, respectively. In conclusion, the observed differences in C(max) and t(max) after i.m. and p.o. administration were consistent with a slower absorption rate of clonazepam after i.m. injection. The systemic exposure to clonazepam was not affected by the route of extravascular administration. Statistical evaluation of these kinetic data showed differences in the absorption rate, so that clonazepam given by the i.m. route is not bioequivalent to the oral route. On the basis of the results of this study, we would recommend the same i.m. and p.o. dose in epileptic patients, but therapeutic response would be expected to be less predictable and to occur later in the case of i.m. administration. PMID- 12646764 TI - Silent infarcts demonstrated by diffusion-weighted MRI in CADASIL. PMID- 12646765 TI - Hepatitis-C-associated multiple arterial dissection of cerebral vessels. PMID- 12646766 TI - Reversible diffusion-weighted MR findings of Salmonella enteritidis-associated encephalopathy. PMID- 12646767 TI - Combined temporal and frontal epileptogenic foci in meningioangiomatosis. PMID- 12646768 TI - LMX1B 17-bp deletion and A3243G mtDNA transition in a previously described patient. PMID- 12646769 TI - Primary CNS mantle cell lymphoma associated with an isolated CSF monoclonal IgG band. PMID- 12646770 TI - Thalamic and mesencephalic hemorrhages after multiple honeybee stings: a life threatening apitherapy complication. PMID- 12646771 TI - Levetiracetam control of myoclonus in a patient with Creutzfeldt-Jakob disease. PMID- 12646772 TI - Oro-mandibular dystonia in a case of multiple sclerosis with capsular plaque. PMID- 12646773 TI - Thrombolytics for cerebral sinus thrombosis: a systematic review. AB - BACKGROUND AND PURPOSE: The use of thrombolytics is frequently mentioned in patients with cerebral venous or dural sinus thrombosis (CVDST) who deteriorate despite anticoagulant therapy. The aim of this review was to collect all the published information about their use in CVDST and to assess their efficacy and safety. METHODS: To find cases of CVDST treated with thrombolytics, we performed a MEDLINE search from 1966 to July 2001, checked all reference lists of studies found and hand searched volumes of 11 journals. Data was extracted by means of a standardised data extraction form. Proportions and 95% confidence intervals (CI) were calculated for outcomes and complications of thrombolytics. Cases were stratified according to variables that may influence the outcome and subgroups were compared by odds ratios and 95% CI. RESULTS: No randomised clinical trial (RCT) was found. Seventy-two studies (169 patients) were included. Urokinase was the thrombolytic most frequently administered (76%). In the majority of cases the thrombolytic was locally infused in the occluded sinus (88%). At discharge, 10 cases (7%; 95% CI 3-12%) were dependent and 9 cases (5%; 95% CI 2-9%) died. Intracranial haemorrhages occurred in 17% of cases. In 5% they were associated with clinical deterioration. Extracranial haemorrhages occurred in 21%, but only 2% required blood transfusion. CONCLUSIONS: Thrombolytics appeared to be reasonably safe in CVDST, but its efficacy cannot be assessed from the published data. Considering that CVDST is an uncommon disease, a randomised controlled trial to assess effectiveness and safety of local thrombolytics in cases of CVDST with poor prognosis is difficult but not impossible to undertake, on a multicentre international collaboration trial. PMID- 12646774 TI - Increased common carotid artery intima media thickness in adolescent hypertension: results from the Debrecen Hypertension study. AB - BACKGROUND: The aim of the present work was to investigate intima media thickness (IMT) in healthy and in hypertensive adolescents and its influencing factors. METHODS: 103 hypertensive and 58 healthy adolescents were studied. IMT was measured in the common carotid artery using B-mode ultrasonography. Additionally, laboratory parameters (blood glucose, cholesterol, triglycerides, HDL- and LDL cholesterols) and left ventricular mass indices were obtained. RESULTS: IMT in the common carotid artery was higher in hypertensive adolescents (means +/- SD: 0.55 +/- 0.11 mm) than in healthy control subjects (0.48 +/- 0.08 mm, p < 0.001). Similarly, a higher left ventricular mass index was measured in hypertensive (103.2 +/- 30.6 g/m(2)) than in healthy teenagers (91.1 +/- 25.2 g/m(2), p < 0.001). In general, IMT in adolescents was associated with age, weight, body mass index, left ventricular mass index and average systolic and diastolic blood pressure values of the subjects. By assessing the multivariate association between IMT and other factors, intima-media thickness was only associated with age and left ventricular mass index of the hypertensive subjects and was independent from all the investigated factors in normotensive controls. CONCLUSIONS: Our data suggest an ongoing target-organ damage in adolescent hypertension. These patients need to be subjected to early diagnostic methods, treatment and a regular follow-up, in order to avoid severe clinical manifestations of secondary target-organ damage due to hypertension. PMID- 12646776 TI - Value of diffusion-weighted imaging in patients with a nonlocalizing examination and vasospasm from subarachnoid hemorrhage. AB - We evaluated the value of diffusion-weighted imaging (DWI) in patients with suspected vasospasm from subarachnoid hemorrhage (SAH). Magnetic resonance imaging (MRI), including DWI, was performed in 5 patients with vasospasm resulting from SAH. Restricted diffusion (ischemia) was present in 3 patients with symptomatic vasospasm. A diffusion abnormality was not demonstrated in 2 patients with asymptomatic vasospasm despite persistently elevated velocities by transcranial Doppler ultrasonography in 1. In this patient, serial MRI studies showed no ischemia. Our results suggest that DWI may provide a method to assess the presence and severity of ischemia in patients with SAH and vasospasm and assist in patient management. PMID- 12646775 TI - Is ultrasound examination sufficient in the evaluation of patients with internal carotid artery severe stenosis or occlusion? AB - BACKGROUND AND PURPOSE: Carotid endarterectomy (CE) has been shown to be beneficial in patients with symptomatic high-grade internal carotid artery (ICA) stenosis. Some authors have suggested that when ultrasound shows a stenosis 70 99%, CE can be performed without further imaging study. However, ultrasound findings that suggest an ICA occlusion, not confirmed by angiography but which instead show a near-occlusion usually benefit from CE. The objectives of this study are: (1). to evaluate how angiography-obtained information on intracranial arteries affects the treatment decision in patients with 70-99% ICA stenosis, and (2). to evaluate when a symptomatic ICA occlusion shown by ultrasound could actually be a patent artery and therefore benefit from CE. MATERIALS AND METHODS: We prospectively collected the cerebral angiograms of 133 consecutive patients with TIA or non-disabling stroke due to large artery disease where ultrasound suggested a stenosis >or=70% or occlusion of symptomatic ICA; we calculated the frequency of intracranial vascular malformations and intracranial artery disease (IAD) located in the infraclinoid or supraclinoid portion of the ICA, and in the anterior or middle cerebral artery. RESULTS: Ultrasound showed 31 ICA occlusions and 102 ICA with 70-99% stenosis. All the patients with an ICA stenosis 70-99% on ultrasound examination had the degree of stenosis confirmed by angiography. Two out of 31 patients did not have a complete occlusion but angiography showed a near-occlusion and consequently they underwent CE. Sixty-five (62.5%) out of 104 patients with patent ICA had IAD (mild 26.9%, moderate 21.2%, and severe 14.4%). Five patients (4.8%) had intracranial vascular malformations (4 aneurysms and 1 arteriovenous malformation). One patient had disabling stroke during angiography. Seven patients (6.7%) did not undergo CE after angiography (1 patient had an aneurysm >10 mm, 1 patient had a very tight stenosis of the basilar artery, 5 patients had intracranial stenosis more severe than the extracranial stenosis). CONCLUSIONS: In patients that on the basis of ultrasound examination can benefit from CE, information on intracranial arteries is necessary. Moreover, complete occlusion cannot be detected with certainty only by ultrasound examination. PMID- 12646777 TI - Visualization of ischaemic penumbra using a computed tomography perfusion method. AB - A computed tomography (CT) perfusion imaging and a diffusion/perfusion magnetic resonance imaging (MRI) were performed in a 51-year-old man at 15 and 15.5 h of onset of left middle cerebral artery infarction, respectively. The established infarct, as revealed by the diffusion-weighted MRI, had low values of cerebral blood flow (CBF) and cerebral blood volume (CBV) on the CT perfusion study. The ischaemic penumbra, as reflected by the perfusion-diffusion mismatch on MRI, had reduced CBF, prolonged mean transit time (MTT) and compensated CBV. Spontaneous bleeding occurred in the medial part of the left lentiform nucleus, immediately following decompressive craniectomy. CT measurements of CBF, CBV and MTT may visualize the ischaemic penumbra and explain the site of bleeding after surgical decompression. PMID- 12646778 TI - Infarct volume on apparent diffusion coefficient maps correlates with length of stay and outcome after middle cerebral artery stroke. AB - BACKGROUND: Diffusion-weighted MRI (DWI) can depict acute ischemia based on decreased apparent diffusion coefficient (ADC) values. ADC maps, unlike DWI (which have contributions from T2 properties), solely reflect diffusion properties. Recent studies indicate that severity of neurological deficit corresponds with degree of ADC alteration. PURPOSE: To determine whether infarct volume on ADC maps correlates with length of hospitalization and clinical outcome in patients with acute ischemic middle cerebral artery (MCA) stroke. STUDY POPULATION: Forty-five consecutive patients with acute (3 SDs below the average ADC value of a contralateral control region. Infarct volume was correlated with length of hospitalization and 6-month outcome assessed with Glasgow Outcome Scale (GOS), Modified Rankin Score (mRS), Barthel Index (BI) and a dichotomized outcome status with favorable outcome defined as GOS 1, mRS or=95. RESULTS: Infarct volume on ADC maps ranged from 0.2 to 187 cm(3) and was significantly correlated with length of hospitalization (p < 0.001, r = 0.67). Furthermore, ADC infarct volume was significantly correlated with GOS (r = 0.73), mRS (r = 0.68), BI (r = 0.67) and outcome status (r = 0.65) (each p < 0.001). Multiple logistic regression revealed a statistically significant correlation between ADC infarct volume and outcome status (p < 0.05), but none for Canadian Neurological Scale score, age and gender (p >0.05 each). CONCLUSION: Infarct volume measured by using a quantitative definition for infarcted tissue on ADC maps correlated significantly with length of hospitalization (as a possible surrogate marker for short-term outcome) and functional outcome after 6 months. ADC infarct volume may provide prognostic information for patients with acute ischemic MCA stroke. PMID- 12646779 TI - Reduced rate of microembolism by optimized aortic cannula position does not influence early postoperative cognitive performance in CABG patients. AB - BACKGROUND AND PURPOSE: The number of microemboli as measured by Doppler ultrasound during coronary artery bypass grafting (CABG) can be reduced by positioning the arterial cannula into the aorta descendens. The aim of this study was to prospectively evaluate whether this alternative aortic cannulation procedure leads to better neuropsychologic outcome early after surgery along with embolus reduction. METHODS: Sixty patients with elective CABG were randomized to either using a short aorta ascendens cannula or an elongated cannula placed in the aorta descendens. All patients were tested by seven neuropsychologic tests preoperatively. Intraoperative embolus detection could be performed by transcranial Doppler in 32 patients. The neuropsychologic tests could be repeated serially until the 9th postoperative day in 54 patients. Patient groups did not differ in terms of preoperative psychometric performance and of the surgical characteristics except cannula positioning. All data were analyzed by a blinded assessor. RESULTS: Neuropsychologic test scores showed in all individual patients a transient decline with subsequent recovery, but did not differ significantly between the groups except for the letter cancellation test at discharge favoring the patients with the longer cannula (102.3 +/- 11.6 vs. 94.5 +/- 11.5 mean +/- SD; p = 0.025). In the subgroup who had Doppler sonography, neuropsychologic test scores did not differ between the groups. However, microembolic signals were markedly reduced in patients with the elongated cannula (median 174.5 vs. 413.0; p = 0.011). CONCLUSIONS: Though reducing brain microembolism, use of an elongated aortic cannula does not appear to influence overall cognitive performance early after CABG in this pilot study. PMID- 12646780 TI - Ischaemic cerebrovascular events in HIV infection: a cohort study. AB - Several case reports and series described ischaemic cerebrovascular events in HIV infection. However, the exact prevalence and the clinical features of these events are unknown. We performed a cohort study on 772 consecutive HIV infected patients and evaluated the rate of transient ischaemic attacks (TIA) and of completed stroke. A total prevalence of 1.9% for TIA (0.8%) and stroke (1.2%) was calculated resulting in an annual incidence rate of 216 per 100000. The prevalence was highest in the later stages of the infection. Stroke patients had a poorer immunological state than the TIA and the cohort patients. Probable (n = 3) and possible (n = 2) vasculitis and cardiogenic embolism (n = 2) could be detected as aetiology, the remaining patients had a cryptogenic event. Our data suggest that ischaemic cerebrovascular events are more common in HIV infected patients than in the general population and that a part of these events might be caused by HIV associated vasculitis or vasculopathy. PMID- 12646781 TI - Distinguishing intracerebral hemorrhages caused by arteriovenous malformations. AB - BACKGROUND: There is a shortage of data addressing the clinical characteristics of patients with arteriovenous malformations (AVMs) who present with intracerebral hemorrhages (ICH). METHODS: A retrospective cohort study of members of a large, pre-paid health care program was conducted to identify factors that distinguish ICH secondary to cerebral AVMs from all other causes. Univariate and multivariate analysis was performed using Student's t test, Wilcoxon rank-sum test, and logistic regression. RESULTS: Patients with an underlying AVM were younger and more likely to be female, non-smokers with lower blood pressures, lower cholesterol, and lower white blood cell counts on presentation. CONCLUSIONS: These clinical characteristics may be useful in defining potential risk factors in future prospective studies as well as targeting candidates for additional imaging studies after ICH with no apparent etiology. PMID- 12646783 TI - Acute inflammatory events and ischemic stroke subtypes. AB - BACKGROUND: Recent studies have suggested that previous infection may be a risk factor for ischemic stroke mainly in young and middle-aged patients. The present study sought to further investigate the association between recent inflammatory events (IE) and ischemic stroke without age restriction and to determine the role of recent IE in different ischemic stroke subtypes. METHODS: We performed a case control study with 93 consecutive hospitalized stroke patients and 200 (107 hospital and 93 community) controls. Acute IE, both infective and non-infective, occurring in the previous 30 days were assessed using a standard questionnaire. The TOAST criteria were used for ischemic stroke subtypes classification. RESULTS: Acute IE in the previous 30 and 7 days were significantly and independently associated with ischemic stroke (37/93 vs. 47/200; OR 2.23, 95% CI 1.26-3.96 and 17/93 vs.16/200; OR 2.45, 95% IC 1.11-5.39, respectively). Stratifying for stroke subtypes, acute IE significantly and independently increased the risk of atherothrombotic (OR 5.72, 95% CI 2.14-15.25) and cardioembolic stroke (OR 3.02, 95%CI 1.20-7.63). CONCLUSIONS: Acute IE increase the risk of acute ischemic stroke of atherothrombotic and cardioembolic type independently of other predisposing factors. Implications for daily clinical practice, in relation to prevention and treatment of IE in patients at risk, have to be explored. PMID- 12646782 TI - Phenylpropanolamine and intracranial hemorrhage risk in a Mexican population. AB - BACKGROUND AND PURPOSE: Phenylpropanolamine (PPA) has been associated with an increased risk of intracranial hemorrhage (ICH). The aim of this study was to assess the association between PPA intake and ICH in a Mexican population. METHODS: We included all patients with ICH aged 18 to 51 years, with no known structural etiology, diagnosed from January 1991 to December 2000. Three to 4 controls per patient matched by sex, age (within 5 years) and place of residence were included. Patients and controls were asked about use of cold medication or appetite suppressant medications within the previous year before the interview. We considered a PPA related hemorrhage when there was a temporal relationship between the use of medication and the development of the hemorrhage, and when other causes could be ruled out. Associated risks for PPA use and other possibly related variables were estimated. RESULTS: 177 patients (mean age 39 +/- 12 years) were included; 58 (33%) were diagnosed with subarachnoid hemorrhage (SAH) and 119 (67%) with ICH. 41.2% (73 of 177) of patients had documented use of PPA within the past year and 10 (5.7%) of them had a temporal relationship between ingestion of PPA and ICH. In control subjects 42.4% (422 of 996) had been exposed to PPA and none of them developed hemorrhage. The time from PPA exposure to the onset of ICH varied from 30 minutes to 24 hours. The risk of PPA exposure for hemorrhage was not significant in cases or controls, OR 0.95 (95% CI, 0.68 to 1.34; p = 0.77). No subjects (cases or controls) reported use of PPA as an appetite suppressant. CONCLUSIONS: We found no association between ingestion of PPA and cerebral hemorrhage with respect to ingestion of PPA in the previous year. When recent use was looked at an apparent risk was evident. PMID- 12646784 TI - Post-transfusion posterior leukoencephalopathy with cytotoxic and vasogenic edema precipitated by vasospasm. PMID- 12646785 TI - Isolation, identification, and quantitation of urinary glycosaminoglycans. AB - BACKGROUND: Substantial amounts of glycosaminoglycans (GAGs) are present in the urine of healthy individuals, but the concentration in the serum is very low. This finding suggests that urinary GAGs come from the glomerulus and may reflect the turnover of GAGs in the glomerulus. HYPOTHESIS: However, little is known about the physiologic regulation of the urinary GAGs in humans, and so investigations are needed to evaluate the effects of age and sex on urinary GAGs in normal individuals. METHODS: Eighty-seven healthy subjects were included in this study. Urinary GAGs were isolated and quantified at the nanogram level by combined azure A-silver staining in agarose gels. RESULTS: The level of urinary GAGs peaked at 10-19 years in both sexes. The proportion of chondroitin sulfate decreased with age, but the proportion of heparan sulfate increased with age. CONCLUSION: The total amount of GAGs and the proportions of chondroitin sulfate, heparan sulfate, and dermatan sulfate appear to change with age. Therefore, investigations in which urinary GAG is used as a parameter of glomerular GAG turnover should ensure that control groups are precisely matched for age. Changes in the proportions of each GAG may be more informative than their absolute levels. PMID- 12646786 TI - Longitudinal monitoring of 2-long terminal repeat circles in peripheral blood mononuclear cells from patients with chronic HIV-1 infection. AB - OBJECTIVE: To evaluate the potential use of 2-long terminal repeats (LTR) HIV circular DNA quantification for the monitoring of ongoing virus replication in treated HIV-1-infected patients. DESIGN AND METHODS: In a longitudinal setting, where the natural course of HIV-1 infection was in most cases disrupted by continuous or discontinuous antiviral therapy, 2-LTR circles of HIV-1 DNA were quantified in serial peripheral blood mononuclear cell samples, selected in retrospect from 16 patients with chronic HIV-1 infection, using quantitative real time PCR. We compared variations of 2-LTR circle level with concomitant variations in plasma viral RNA level and with the frequency of productively infected cells and chromosome associated proviral DNA copy numbers in patient's peripheral blood mononuclear cells. RESULTS: Antiviral treatment led to a sharp decrease in plasma viraemia and infectious cell frequency. In contrast, we found that levels of proviral DNA and 2-LTR circles were significantly lower under treatment only when groups of specimens that were homogeneous, with respect both to plasma viraemia and infectious cell frequency, were compared. Moreover, during the time of undetectable plasma viraemia, scarcely any decline in proviral DNA or 2-LTR circle levels was observed. CONCLUSIONS: The low impact of antiviral treatment on 2-LTR circle levels in vivo, when plasma viraemia and infectious cell frequency both dramatically decline lead us to conclude that 2-LTR circles should not be used for the monitoring of recent viral replication in treated patients. PMID- 12646787 TI - Blocking of cell-free and cell-associated HIV-1 transmission through human cervix organ culture with UC781. AB - OBJECTIVE: To determine the efficacy of UC781 in preventing HIV-1 transmission through cervical tissue. DESIGN: Use of human cervical tissue organ culture, in which the cervix is in the upper chamber of a transwell and transmission of infective virus is quantified in the lower chamber. METHODS: Five-millimeter pieces of cervical tissues are exposed to UC781. After thorough removal of the drug, the tissues are exposed to high doses of cell-free or cell-associated HIV 1. Transmission of HIV-1 through the cervix is measured by determining infection of target cells in the lower chamber. RESULTS: Exposure of cervix to 0.5 microM UC781 for 30 min, followed by extensive washing away of the residual drug, resulted in 95% reduction of subsequent viral transmission. Exposure of the cervix to 1 microM UC781 for 20 min, or 10 microM UC781 for 2 min, resulted in neutralization of T- and M-tropic HIV-1 isolates of various clades, and prevention of cell-associated HIV-1 transmission. Moreover, a 20 min incubation with 10 microM UC781 abolished HIV-1 transmission through the cervix for 48 h after drug pretreatment. Importantly, UC781 was not toxic, even when the cervical tissues were exposed to 20 microM UC781 for 24 h. UC781 was effective against transmission of both cell-free and cell-associated HIV-1 also when formulated into a non-spermicidal (Replen) or spermicidal (BufferGel) gel. CONCLUSIONS: Exposure of the cervix to UC781 results in blocking of subsequent HIV-1 transmission with no toxicity. Therefore, UC781 is an excellent candidate microbiocide. PMID- 12646788 TI - Processivity and drug-dependence of HIV-1 protease: determinants of viral fitness in variants resistant to protease inhibitors. AB - OBJECTIVE: To investigate the role of processivity and drug-dependence of HIV-1 protease as fitness determinants in variants resistant to protease inhibitors (PI). DESIGN AND METHODS: HIV-1 protease sequences from 32 infected subjects (27 patients who failed PI-treatments and five PI-naive controls) were evaluated using a recombinant method. The HIV-1 phenotype to seven PI was analysed together with the replication capacity of recombinants and the processivity and drug dependence of the HIV-1 proteases. Protease mutants (positions 10, 46, 54, 82, 84, 90, and combinations thereof) were generated in vitro and studied under identical experimental conditions. RESULTS: In the absence of PI, 24 of 27 (89%) resistant proteases from treated subjects showed decreased processivity compared with the wild type. Processivity was lower in sequences bearing fewer mutations, than in more mutated ones. Twelve sequences (44%) conferred slower replication kinetics to the recombinant viruses. Seven sequences (26%) showed higher processivity levels in the presence of PI than in their absence, suggesting that drug-dependence influences PI-resistant variants. Among the mutants generated in vitro, mutations 82A and 90M determined broad cross-resistance to PI in association with 10I. A drop of processivity was observed for the 82A+90M variants; 10I allowed partial recovery for 82A and 84V, and marked recovery for 90M mutants. CONCLUSIONS: A decrease in HIV-1 protease processivity parallels early selection of primary mutations, whereas its recovery is driven by compensatory mutations. Furthermore, a PI may select drug-dependent, besides resistant, HIV-1 protease variants. Changes in processivity and drug-dependence of HIV-1 proteases have implications in the replication capacity of PI-resistant viruses. PMID- 12646789 TI - Persistent non-gastrointestinal metabolic acidosis in pediatric HIV-1 infection. AB - OBJECTIVES: To determine the incidence and to identify the clinical parameters associated with non-gastrointestinal renal tubular and high anion gap acidosis in a cohort of HIV-1-infected children. METHODS: Records of 202 HIV-1-infected children were reviewed to identify patients with metabolic acidosis. Serum and urine chemistries of those children with persistent non-gastrointestinal acidosis were then studied prospectively. Serum and urinary anion gaps (SAG and UAG) were calculated. Those with acidosis (group 1) were compared with children without acidosis (group 2). Associations were determined with Pediatric HIV classification, height, weight, antiretroviral therapy, and Pneumocystis carinii pneumonia prophylaxis. RESULTS: Persistent acidosis was noted in 34 out of 202 children (17%): 16 out of 34 (47%, group 1A) had elevated SAG acidosis, and 18 out of 34 (53%) had normal SAG acidosis with a positive UAG (distal renal tubular) acidosis (group 1B). Those with acidifying defects more often received P. carinii pneumonia prophylaxis (P = 0.02 and 0.01 for groups 1 and 1A, respectively) independently of HIV-1 classification. This group was shorter in height than group 2 (P = 0.007). Differences in weight were not significant (P = 0.1). However, acidotic subjects were more immunocompromised than those in group 2 (multivariate P < 0.001 for HIV classification C3). CONCLUSIONS: Elevated SAG acidosis and renal tubular acidosis are not uncommon among HIV-infected children with advanced disease. These disorders may be associated with height growth failure and prophylaxis with sulfur/sulfone containing antibiotics. HIV infection and/or its associated therapies may cause renal tubular damage. The causes of elevated SAG acidosis require further investigation. PMID- 12646790 TI - Identification of two mutually exclusive groups after long-term monitoring of HIV DNA 2-LTR circle copy number in patients on HAART. AB - BACKGROUND: Anti-retroviral drug therapy reduces but does not abolish HIV transmission and replication throughout the body. HIV DNA 2-long terminal repeat (2-LTR) circles have been shown in point-based studies to persist in some patients whose plasma HIV RNA was undetectable. However, the degree of fluctuation of circle copy number over time has not been determined. METHODS: A reliable, reproducible and robust quantitative LightCycler (LC qPCR)-based assay for HIV DNA 2-LTR circles in peripheral blood mononuclear (PBMN) cells was established. A prospective, longitudinal study of these circles was undertaken in HIV-1-positive patients on anti-retroviral therapy whose plasma HIV RNA was undetectable at < 50 copies/ml. Patients starting therapy for the first time were also monitored. RESULTS: A cohort of 60 patients whose plasma HIV RNA was undetectable for 32 +/- 2 months were monitored for circles for 15 +/- 2 months. The circle copy number ranged from < 10 to 620 copies/106 PBMN cells. The circle negative (< 10 copies/1 x 106 PBMN) cells group of 36 patients and the circle positive (> 10 copies/106 PBMN cells) group of 24 patients were mutually exclusive (P < 0.0001). The mean circle half-life in seven of the 10 patients starting anti-retroviral therapy for the first time was 5.7 days. CONCLUSION: The circle assay is useful for identifying those patients in whom transmission of infectious virus continues despite prolonged periods of time during which plasma HIV RNA is undetectable. New drug combinations and new therapeutic approaches should be aimed at those patients whose plasma HIV RNA is undetectable but who remain positive for 2-LTR circles. PMID- 12646791 TI - Conformational changes in env oligomer induced by an antibody dependent on the V3 loop base. AB - OBJECTIVE: The HIV-1 env oligomer is structured such that conserved, neutralizing epitopes are obscured by gp120 variable loops. We have studied the ability of an IgG2 human monoclonal antibody (hmAb), F425 B4e8 (B4e8), dependent upon the base of the V3 loop, to induce conformational changes in the env oligomer. DESIGN: The effect of B4e8 antibody on the exposure of neutralizing epitopes and viral neutralization was studied in combination with other hmAb. METHODS: Epitope exposure and viral neutralization was determined using native, intact primary isolate virions. RESULTS: B4e8 antibody neutralizes infection and binds to HIV infected cells and primary isolate virions. B4e8 and 2G12 enhanced the binding of each other to infected cells or virus and the combination resulted in synergistic neutralization. B4e8 also enhanced the binding of CD4i and CD4 binding site antibodies. CONCLUSIONS: The conserved epitopes exposed by B4e8 are similar to those exposed by the movement of the variable loops following CD4 engagement. Further studies with select antibody combinations should provide important information for the design of effective immunotherapeutic agents. PMID- 12646792 TI - A phase II clinical study of the long-term safety and antiviral activity of enfuvirtide-based antiretroviral therapy. AB - OBJECTIVES: The primary objective was to determine the long-term safety of the subcutaneous self-administration of enfuvirtide. Secondary objectives included the determination of enfuvirtide pharmacokinetics and antiviral activity and the immunological response to the enfuvirtide-containing regimen. METHODS: A multicenter 48-week uncontrolled open-label rollover study was conducted on 71 HIV-infected adults recruited from previous enfuvirtide clinical trials. Patients with extensive previous use of protease and reverse transcriptase inhibitors received a twice-daily dose of 50 mg enfuvirtide subcutaneously (45 mg deliverable) combined with two or more antiretroviral drugs selected for each individual, guided by resistance testing and previous treatment history. RESULTS: The mean baseline plasma HIV-RNA level was 4.81 log(10) copies/ml and the mean CD4 cell count was 134.8 cells/microl. The majority (86.9%) of treatment-emergent adverse events were grade 2 or less in severity. Injection site reactions were common, but no patients discontinued treatment. A mean HIV-RNA change of -1.33 log(10) was achieved within 14 days of treatment initiation. At week 48, approximately one-third of all patients in the intent-to-treat population maintained significant suppression of plasma HIV RNA, with either less than 400 copies/ml or more than a 1.0 log(10) decline from baseline. The mean gain in absolute CD4 cell counts at 48 weeks was 84.9 cells/microl. Trough plasma concentrations of enfuvirtide were consistently higher than target concentrations. CONCLUSION: Self-administration of enfuvirtide is not associated with unexpected toxicities for up to one year, and combined with oral antiretroviral drugs was associated with a significant decrease in HIV RNA and an increase in CD4 cell counts. PMID- 12646793 TI - Double-blind, randomized, placebo-controlled phase III trial of oxymetholone for the treatment of HIV wasting. AB - BACKGROUND: Despite highly active antiretroviral therapy (HAART), chronic involuntary weight loss still remains a serious problem in the care of HIV patients. Various alterations in energy metabolism and endocrine regulation have been found to cause loss of lean body mass (LBM) and body cell mass (BCM). Previous studies in HIV-positive men undergoing androgen replacement therapy or treatment with recombinant growth hormone (rGH) have shown partial restoration of LBM, but these treatments have largely been ineffective in eugonadal individuals. STUDY DESIGN: Double-blind, randomized, placebo-controlled trial of 89 HIV positive women and men with wasting assigned to the anabolic steroid oxymetholone [50 mg twice (BID) or three times daily (TID)] or placebo for 16 weeks followed by open-label treatment. STUDY ENDPOINTS: Body weight, bioimpedance measurements, quality of life parameters and appetite. RESULTS: Oxymetholone led to a significant weight gain of 3.0 +/- 0.5 and 3.5 +/- 0.7 kg in the TID and BID groups, respectively (P < 0.05 for each treatment versus placebo), whereas individuals in the placebo group gained an average of 1.0 +/- 0.7 kg. Body cell mass increased in the oxymetholone BID group (3.8 +/- 0.4 kg; P < 0.0001) and in the oxymetholone TID group (2.1 +/- 0.6 kg; P < 0.005), corresponding to 12.4 and 7.4% of baseline BCM, respectively. Significant improvements were noted in appetite and food intake, increased well-being and reduced weakness by self examination. The most important adverse event was liver-associated toxicity. Overall, 35% of patients in the TID, 27% of patients in the BID oxymetholone group and no patients in the placebo group had a greater than five times baseline increase for alanine aminotransferase during the double-blind phase of the study. CONCLUSIONS: Oxymetholone can be considered an effective anabolic steroid in eugonadal male and female patients with AIDS-associated wasting. The BID (100 mg/day) regimen appeared to be equally effective as the TID (150 mg/day) regimen in terms of weight gain, LBM and BCM and was associated with less, but still significant liver toxicity. PMID- 12646794 TI - Is there a baseline CD4 cell count that precludes a survival response to modern antiretroviral therapy? AB - OBJECTIVE: Therapeutic guidelines advise that 200-350 x 106 cells/l may approximate an irreversible threshold beyond which response to therapy is compromised. We evaluated whether non-immune-based factors such as physician experience and adherence could affect survival among HIV-infected adults starting HAART. METHODS: Analysis of 1416 antiretroviral naive patients who initiated triple therapy between 1 August 1996 and 31 July 2000, and were followed until 31 July 2001. Patients whose physicians had previously enrolled six or more patients were defined as having an experienced physician. Patients who received medications for at least 75% of the time during the first year of HAART were defined as adherent. Cumulative mortality rates and adjusted relative hazards were determined for various CD4 cell count strata. RESULTS: Among patients with < 50 x 106 cells/l the adjusted relative hazard of mortality was 5.07 [95% confidence interval (CI), 2.50-10.26] for patients of experienced physicians and was 11.99 (95% CI, 6.33-22.74) among patients with inexperienced physicians, in comparison to patients with > or = 200 x 106 cells/l treated by experienced physicians. Similarly, among patients with < 50 x 106 cells/l, the adjusted relative hazard of mortality was 6.19 (95% CI, 3.03-12.65) for adherent patients and was 35.71 (95% CI, 16.17-78.85) for non-adherent patients, in comparison to adherent patients with > or = 200 x 106 cells/l. CONCLUSION: Survival rates following the initiation of HAART are dramatically improved among patients starting with CD4 counts < 200 x 106 cells/l once adjusted for conservative estimates of physician experience and adherence. Our results indicate that the current emphasis of therapeutic guidelines on initiating therapy at CD4 cell counts above 200 x 106 cells/l should be re-examined. PMID- 12646795 TI - Serum biochemical markers accurately predict liver fibrosis in HIV and hepatitis C virus co-infected patients. AB - OBJECTIVE: Liver biopsy, the gold standard for assessing hepatitis C virus (HCV) related fibrosis, is invasive and prone to complications. Our aim was to determine the operating characteristics of a non-invasive index of biochemical markers for the prediction of fibrosis in patients with HIV/HCV co-infection. DESIGN: In a cross-sectional, cohort study in a French tertiary-care hospital 130 HIV/HCV-co-infected patients with a liver biopsy and serum were tested for markers of liver fibrosis. METHODS: An index incorporating age, sex, alpha(2) macroglobulin, apolipoprotein A1, haptoglobin, bilirubin, and gamma-glutamyl transpeptidase (GGT), derived using multivariate logistic regression, was compared with liver histology. HIV-specific indices including the CD4 cell count and HIV-RNA load were also constructed. The diagnostic values of the indices were compared using receiver operating characteristic (ROC) curves. MAIN OUTCOME MEASURE: Septal fibrosis (F2-F4) by the METAVIR classification. RESULTS: By multivariate analysis, the most informative markers were alpha(2)-macroglobulin, apolipoprotein A1, GGT, and sex. The area under the ROC curve of the five-marker index was 0.856 +/- 0.035; not significantly different from the HIV-specific indices. On a scale from zero to 1.00, the five-marker index had a positive predictive value of 86% for scores greater than 0.60, and a negative predictive value of 93% for scores of 0.20 or less. These thresholds could reduce the necessity for liver biopsy by 55% while maintaining an accuracy of 89%. CONCLUSION: An index including five biochemical markers accurately predicts significant fibrosis in patients with HIV/HCV co-infection, and may substantially reduce the necessity for liver biopsy. PMID- 12646796 TI - Safety and acceptability of the Reality condom for anal sex among men who have sex with men. AB - OBJECTIVES: To assess safety and acceptability of Reality condoms for anal sex among men who have sex with men. METHODS: Crossover study among HIV seroconcordant (33 HIV-negative and 5 HIV-positive) monogamous male couples, randomized to latex male and Reality condom use with anal sex. RESULTS: Slippage with removal was reported more frequently with Reality than male latex condoms [odds ratio (OR), 2.7; 95% confidence interval (CI), 1.2-5.8 for receptive partners and OR, 34.1; 95% CI, 13.8-84.1 for insertive partners]. Receptive partners more frequently reported pain or discomfort (OR, 5.0; 95% CI, 2.6-9.4) and rectal bleeding (OR, 1.9; 95% CI, 0.9-4.1) with Reality condoms than male condoms. Over 20% reported willingness to use the Reality condom in the future with a partner of unknown HIV status; willingness was associated with past problems with male condoms and no problems with Reality condoms among receptive partners, and with past use of Reality condoms and HIV seropositivity among insertive partners. CONCLUSIONS: Men reported more frequent problems with Reality condoms than male latex condoms used for anal intercourse, particularly slippage, discomfort, and rectal bleeding. Design modifications, training, and research on the clinical significance of safety outcomes are needed for use of Reality condoms with anal sex. PMID- 12646797 TI - Sexual behavior of HIV discordant couples after HIV counseling and testing. AB - BACKGROUND AND OBJECTIVES: Sexual behavior following voluntary HIV counseling and testing (VCT) is described in 963 cohabiting heterosexual couples with one HIV positive and one HIV negative partner ('discordant couples'). Biological markers were used to assess the validity of self-report. METHODS: Couples were recruited from a same-day VCT center in Lusaka, Zambia. Sexual exposures with and without condoms were recorded at 3-monthly intervals. Sperm detected on vaginal smears, pregnancy, and sexually transmitted diseases (STD) including HIV, gonorrhea, syphilis, and Trichomonas vaginalis were assessed. RESULTS: Less than 3% of couples reported current condom use prior to VCT. In the year after VCT, > 80% of reported acts of intercourse in discordant couples included condom use. Reporting 100% condom use was associated with 39-70% reductions in biological markers; however most intervals with reported unprotected sex were negative for all biological markers. Under-reporting was common: 50% of sperm and 32% of pregnancies and HIV transmissions were detected when couples had reported always using condoms. Positive laboratory tests for STD and reported extramarital sex were relatively infrequent. DNA sequencing confirmed that 87% of new HIV infections were acquired from the spouse. CONCLUSIONS: Joint VCT prompted sustained but imperfect condom use in HIV discordant couples. Biological markers were insensitive but provided evidence for a significant under-reporting of unprotected sex. Strategies that encourage truthful reporting of sexual behavior and sensitive biological markers of exposure are urgently needed. The impact of prevention programs should be assessed with both behavioral and biological measures. PMID- 12646798 TI - The cost effectiveness of universal antenatal screening for HIV in New Zealand. AB - OBJECTIVE: To model the incremental costs and benefits of a universal antenatal HIV screening programme in New Zealand (NZ). DESIGN: Cost effectiveness analysis, including only health service costs, using secondary data sources and expert opinion. Uncertainty assessed in multi-way sensitivity analyses. SETTING: The NZ Health Care System. SUBJECTS: Antenatal population of NZ. INTERVENTION: Universal antenatal HIV screening programme. MAIN OUTCOME MEASURES: Incremental cost per true-positive HIV case detected in mothers; incremental cost per HIV case avoided in babies; and incremental cost per discounted life-year gained, for mothers and babies, due to screening. RESULTS: Using base case values the application of universal screening would cost an additional $NZ 723 607 ($US 307 917) and would lead to the identification of an additional 6.25 true-positive women. After terminations have been excluded, the screening programme would detect five HIV exposed babies. There would be 1.15 avoided cases of HIV infection in babies and a net gain of 41.97 discounted life-years, for mothers and babies combined. The cost per incremental HIV-positive woman detected was $NZ 115 859 ($US 49 301), HIV infected baby avoided $NZ 629 669 ($US 267 944) and discounted life-year gained $NZ 17 241 ($US 7336). CONCLUSION: The discounted cost per life gained in NZ compares favourably to estimates reported in studies of similar interventions in other developed countries and other health care interventions in NZ. The decision of whether to implement universal screening in NZ would be clarified if the prevalence of antenatal HIV infection was known and policy makers identified their willingness to pay for an additional life-year gained. PMID- 12646799 TI - Measuring covert HIV replication during HAART: the abundance of 2-LTR circles is not a reliable marker. PMID- 12646800 TI - Treatment of chronic hepatitis C virus infection: we must target the virus or liver fibrosis? PMID- 12646801 TI - The economic profile of antenatal HIV testing: pharmaceutical and methodological considerations. PMID- 12646802 TI - Lack of evidence for an association between a polymorphism in CX3CR1 and the clinical course of HIV infection or virus phenotype evolution. PMID- 12646803 TI - Human cytomegalovirus glycoprotein N genotypes in AIDS patients. PMID- 12646804 TI - Thyroid hormones in HIV-infected patients in the highly active antiretroviral therapy era: evidence of an interrelation between the thyroid axis and the immune system. PMID- 12646805 TI - Increased soluble markers of endothelial dysfunction in HIV-positive patients under highly active antiretroviral therapy. PMID- 12646806 TI - Viral breakthrough after suppression with highly active antiretroviral therapy: experience from 233 individuals with viral loads of less than 50 copies/ml followed for up to 4 years. PMID- 12646807 TI - Glitazones in lipodystrophy syndrome induced by highly active antiretroviral therapy. PMID- 12646808 TI - Small dense LDL and atherogenic lipid profile in HIV-positive adults: influence of lopinavir/ritonavir-containing regimen. PMID- 12646809 TI - Determinants of HIV serconversion in an era of increasing HIV infection among young gay and bisexual men. PMID- 12646810 TI - MBL2 polymorphisms are involved in HIV-1 infection in Brazilian perinatally infected children. PMID- 12646811 TI - Sorting out the complexities of HIV-1 fitness. PMID- 12646812 TI - Failure to detect a non-B HIV-1 subtype by the HIV-1 Amplicor Monitor test, version 1.5: a case of unexpected vertical transmission. PMID- 12646813 TI - Rapid hepatitis B virus-DNA decay in co-infected HIV-hepatitis B virus 'e-minus' patients with YMDD mutations after 4 weeks of tenofovir therapy. PMID- 12646815 TI - Universal access to antiretroviral therapy may be the best approach to 'Do no harm' in developing countries: the Brazilian experience. PMID- 12646814 TI - Quantitative markers for cytomegalovirus disease in HIV-infected patients receiving highly active antiretroviral therapy. PMID- 12646816 TI - Pregnancies among high-risk women in the first ever HIV vaccine efficacy trial. PMID- 12646817 TI - Fulminant Kaposi's sarcoma without mucocutaneous disease. PMID- 12646818 TI - A combined adult health/mental health clinical experience: fostering holistic practice. PMID- 12646819 TI - Determining learning style preferences of students. PMID- 12646820 TI - Activities designed to socialize students into nursing. PMID- 12646821 TI - Student drawing: a clinical learning tool. PMID- 12646822 TI - The use of humor and role-playing in reinforcing key concepts. PMID- 12646824 TI - Video streaming in nursing education: bringing life to online education. AB - Distance education is a standard form of instruction for many colleges of nursing. Web-based course and program content has been delivered primarily through text-based presentations such as PowerPoint slides and Web search activities. However, the rapid pace of technological innovation is making available more sophisticated forms of delivery such as video streaming. High quality video streams, created at the instructor's desktop or in basic recording studios, can be produced that build on PowerPoint or create new media for use on the Web. The technology required to design, produce, and upload short video streamed course content objects to the Internet is described. The preparation of materials, suggested production guidelines, and examples of information presented via desktop video methods are presented. PMID- 12646823 TI - Making nursing research "real": an experiential approach. AB - Research can be a challenging subject for nursing students at all levels. Although a research course was a prerequisite for admittance to our masters of science in nursing program at Viterbo University, faculty found that students had varying levels of expertise with the content as well as differing attitudes toward research in practice. To initiate "research thinking" early in the program, 2 faculty members integrated research concepts across 2 of the core courses, one being the nursing research course. Both quantitative and qualitative methods were taught using didactic and experiential methods over 2 semesters. Through the combined activities in the 2 courses, many skills and a new appreciation for nursing research were instilled. PMID- 12646825 TI - Teaching by twenties: mini-lectures about infectious diseases and bioterroristic agents. PMID- 12646826 TI - Strategies for teaching loss, grief, and bereavement. AB - Teaching loss, grief, and bereavement to nursing students should be an interactive process to stimulate critical thinking and address the affective domain of learning. Lecture as a teaching methodology may be the easiest to prepare and deliver; however, used alone, it is ineffective in identifying perceptions, fears, and issues related to dying and death. Personal and professional experiences of loss, grief, and bereavement are central to student's learning of effective and compassionate care of the dying patient and their family. Strategies that explore such experiences allow students to move forward and focus on the cognitive retention of content related to loss, grief, and bereavement, as well as the ability to learn related psychomotor skills. The authors discuss pedagogical methods for teaching student nurses about loss, grief, and bereavement utilizing the End of Life Nursing Education Consortium (ELNEC) curriculum training materials. PMID- 12646827 TI - The promise and perils of a weekend nursing program. AB - In 1996, a weekend nursing program was created as an addition to the traditional weekday schedule to accommodate an increased number of qualified applicants seeking admission to the baccalaureate nursing program. Five years after the inception of the program, a comprehensive evaluation study was conducted to examine the impact of the weekend course schedule on students and faculty. The opportunities and challenges associated with the implementation of an alternative program schedule for entry-level nursing students are presented. PMID- 12646828 TI - A case for problem-based collaborative learning in the nursing classroom. AB - To facilitate student learning of the content in a clinical nursing course, the authors used ideas from Problem-based Learning (PBL) and collaborative learning to develop student writing assignments and group workshops. They describe the PBL enhanced activities and student work, concluding with student feedback and future directions for this curriculum. PMID- 12646829 TI - Promoting personal safety in community health: four educational strategies. AB - Nurse educators must confront the issue of nursing students' personal safety in community health practice, provide the necessary knowledge and skills, and document their actions. Learning opportunities are required to assist students to recognize and evaluate risks, and prevent their consequences. Thematic analysis of interviews with 56 public health nurses in Alberta, Canada uncovered knowledge, skill, and attitudinal components of risk assessment. Categories under the theme framing personal risk in work environments and the sub-themes framing for action and framing for no action provided a conceptual framework for planning educational strategies. Based on how these nurses framed their personal risk, 4 educational strategies are presented: a checklist, small group learning activities, a template for Web-based courseware, and a problem-based learning scenario. PMID- 12646830 TI - Collaborative testing: an innovative approach to test taking. AB - Collaborative testing was used with second semester associate degree nursing students during their fluid and electrolyte content to enhance collaboration and joint problem solving. Student nurses were randomly assigned a partner and allowed to change answers to test questions for 10 minutes at the end of the exam. Outcomes included an increase in student comprehension and test scores with decreased test anxiety. PMID- 12646831 TI - Child abuse and the otolaryngologist: part I. AB - Nearly 1 million infants and children are neglected and abused yearly in the United States, with a greater than 1% resulting mortality rate. One half of these children are seen by physicians for abuse-related injuries, and nearly 75% have injuries of the head and neck. Physicians, however, account for reporting only 11% of all cases. As experts trained in diseases and injuries of the head and neck, otolaryngologists are particularly well positioned to recognize abuse in the clinic and in the emergency room and during other consultations. We present an overview of child abuse definitions, risk factors, and legal obligations of the physician. We also review the manifestations of child abuse within the head and neck, with particular attention to the role of the otolaryngologist. We briefly discuss some conditions that may be mistaken for abuse and suggest a practical protocol for management of suspected cases in the clinic. PMID- 12646832 TI - Child abuse and the otolaryngologist: part II. PMID- 12646833 TI - Harmonic scalpel tonsillectomy in children: a randomized prospective study. AB - OBJECTIVE: In this randomized prospective study, we evaluated postoperative morbidity after use of the Harmonic Scalpel (HS), an ultrasonic dissector coagulator (Ethicon Endo-Surgery Inc, Cincinnati, OH), or conventional electrocautery (EC) during tonsillectomy. STUDY DESIGN AND SETTINGS: Pediatric subjects at 2 sites were randomized and underwent tonsillectomy. Intraoperative blood loss and operation duration were recorded. Postoperative parameters and complications were recorded. RESULTS: One hundred seventeen subjects completed the study. For the HS group, mean operative time was significantly longer (P < 0.001), but intraoperative blood loss was equivalent (P = 1.000). HS subjects slept soundly on postoperative days 1, 2, 3, and 14 (P = 0.041, 0.013, 0.022, and 0.038, respectively, compared with EC group). Mean postoperative pain scores trended lower for HS subjects on postoperative days 2, 3, and 4. CONCLUSION: The use of the HS in pediatric tonsillectomy showed no increase in intraoperative or postoperative blood loss compared with the use of EC, and HS provided possible clinical advantages over EC in patient comfort. SIGNIFICANCE: Tonsillectomy subjects in the HS group showed a statistically significant ability to sleep soundly, suggesting that the subjects experienced less pain. These data correlate with the observed decrease in pain scores. PMID- 12646834 TI - Revisiting outpatient tonsillectomy in young children. AB - OBJECTIVE: Postoperative same-day discharge is safe for most children undergoing tonsillectomy. However, young children with upper airway obstruction have a higher risk of postoperative complications. We review our tonsillectomy experience in children under 36 months to evaluate the safety of outpatient tonsillectomy in this population. STUDY DESIGN AND SETTING: We conducted a retrospective study of all children under 36 months who underwent tonsillectomy during a recent 2-year period at The Children's Hospital of Philadelphia. RESULTS: The indication for tonsillectomy in 96% of 421 children was upper airway obstruction. Eighteen percent required postoperative treatment to prevent respiratory compromise; 56% of these patients had no associated medical comorbidity. Patients younger than 24 months and those with medical comorbidities were more likely to require intervention. CONCLUSION AND SIGNIFICANCE: It is not possible to preoperatively anticipate which children will have postsurgical complications. We recommend planning an overnight admission for children younger than 36 months undergoing tonsillectomy. PMID- 12646835 TI - An evidence-based review of the treatment of peritonsillar abscess. AB - OBJECTIVE: Peritonsillar abscess (PTA) is commonly seen but still controversial. We performed an evidence-based review to answer 3 questions: Are steroids beneficial? Which is the best technique for acute surgical management? When is tonsillectomy indicated? STUDY DESIGN: We performed a MEDLINE search of the published literature using appropriate search terms to identify pertinent articles, which were reviewed and graded according to the evidence quality. RESULTS: Forty-two articles were analyzed. There are no published studies on steroids in PTA. There were 5 level I clinical studies on surgical technique, which indicated that needle aspiration, incision and drainage, and quinsy tonsillectomy are all effective for initial management. The overall PTA recurrence rate is 10% to 15%. CONCLUSIONS: Overall, grade C evidence indicates that several methods of initial surgical drainage are equally effective, and the recurrence rate is low. The literature does not specifically address different treatments for children and adults. PMID- 12646836 TI - Evaluation of sleep-disordered breathing in children. AB - OBJECTIVE: We evaluated changes in disease-specific quality of life (QOL) for children with sleep-disordered breathing (SDB). PATIENTS, DESIGN, AND SETTING: We conducted a cohort study in 69 children (age range, 6 months to 13 years; median age, 6.1 years) from a hospital-based pediatric otolaryngology practice in a metropolitan area. We used the OSA-18, an 18-item QOL survey with known test retest reliability, internal consistency, and construct validity. Survey domains included sleep disturbance, physical suffering, emotional distress, daytime problems, and caregiver concerns. INTERVENTION: Tonsillectomy or adenoidectomy was performed as part of routine clinical care. The OSA-18 was completed at entry by the child's caregiver and again at least 4 weeks after surgery. MAIN OUTCOME MEASURES: The validity of the OSA-18 change score was assessed by comparison with multiple external constructs, including a validated measure of longitudinal change (OSD-6 survey). Responsiveness was assessed by the standardized response mean (SRM). RESULTS: The mean baseline OSA-18 survey score was 3.1 (SD, 0.9) of a maximum 7.0, with higher values indicating poorer QOL. Change scores were available for all children, with a mean interval between preoperative and postoperative surveys of 61 days (range, 42 to 92 days). The mean OSA-18 change score of 1.14 (SD, 0.71) showed significant correlation with tonsil size (r = 0.40), adenoid size (r = 0.31), OSD-6 change scores (r = 0.71), and the direct caregiver estimates of change (r = 0.34). OSA-18 change scores showed large responsiveness to change (SRM 1.44; 95% confidence interval, 1.16 to 69), with the most responsive domains being sleep disturbance, physical suffering, and caregiver concerns. Significant responsiveness was also demonstrated for the domains of daytime problems and emotional distress. CONCLUSIONS: The OSA-18 is a valid, reliable, and responsive QOL measure that combines the advantages of a discriminative and evaluative survey in a single instrument. The OSA-18 is a useful measure of patient-based outcomes for clinical trials, health services research, and routine clinical care. PMID- 12646837 TI - Nonattended home automated continuous positive airway pressure titration: comparison with polysomnography. AB - OBJECTIVE: Automatic adjusting nasal continuous positive airway pressure titration (APAP) has been introduced as an alternative method of establishing pressures for patients with sleep apnea. The performance and accuracy of APAP in nonattended home environment are controversial. This study assessed APAP polysomnographic outcomes and accuracy in a nonattended home environment. STUDY DESIGN AND SETTING: We conducted a retrospective consecutive case series of 24 consecutive patients who had nonattended APAP and simultaneous full polysomnography (PSG). RESULTS: APAP was tolerated and reduced obstructive Apnea Hypopnea Index (AHI) to <10 events/h in all patients. Mean AHI decreased from 38.4 (21.2) to 5.9 (6.6) events/h. Central apneas worsened in one patient. A therapeutic pressure was determined in 91% of patients. Autoset accurately measured residual AHI compared with PSG (R = 0.77, P < 0.001). APAP overestimated the AHI by 1.4 events/h. CONCLUSIONS: Nonattended APAP is successful in many patients in determining a therapeutic positive pressure setting. Reported AHI via Autoset is similar to that of PSG. SIGNIFICANCE: APAP reduces AHI and is tolerated in a nonattended environment. PMID- 12646838 TI - Percutaneous dilation tracheotomy versus surgical tracheotomy: our experience. AB - BACKGROUND: Percutaneous dilation tracheotomy (PDT) is becoming a popular alternative to surgical tracheotomy. In our department, we recently adopted the use of the PDT in intensive care unit patients. Here, we compare the results of the use of these 2 techniques on 150 patients, all performed by the same surgeon. We discuss the pros and cons of PDT and present our experience with the technique compared with surgical tracheotomy (ST). MATERIALS AND METHODS: A prospective study of 75 PDTs and a retrospective study of 75 surgical tracheotomies (ST) were performed at the Department of Otolaryngology-Head and Neck Surgery, Rambam Medical Center, Haifa, Israel. Age, sex, duration of intubation before surgery, time interval between the decision to perform and the performance of tracheotomy, and cost were compared. RESULTS: One hundred fifty tracheotomies were reviewed. The indication for tracheotomy in both groups was prolonged mechanical ventilation. Seven patients were found unsuitable for PDT and underwent ST. Complications included 3 cases of mild postoperative hemorrhage in the ST group, and 1 case of subcutaneous emphysema, 1 case of stomal cellulitis and 2 cases of mild postoperative hemorrhage in the PDT group. The average waiting interval was between 2 to 5 days for ST and 1 to 24 hours for PDT. The intraoperative time for ST was 20 minutes; for PDT, 5 minutes. The cost was 565 dollars for ST and 274 dollars for PDT. CONCLUSIONS: PTD provides an easy, less expensive, and convenient alternative to ST and should be added to the otolaryngologists' armamentarium of surgical airway procedures. The procedure is advantageous for the patient. Complication rates of both techniques are similar and low; however, PDT is a blind technique of obtaining a surgical airway and therefore holds more potential for serious complications. It is our conclusion that this technique is suitable for many, but not all, critical care patients and that the procedure should be performed only by surgeons who are capable of urgently obtaining a surgical airway or exploring the neck should the PDT fail. PMID- 12646839 TI - Gene discovery using a human vestibular schwannoma cDNA library constructed from a patient with neurofibromatosis type 2 (NF2). AB - BACKGROUND: Despite a strong association of schwannomin/merlin gene mutations with vestibular schwannoma formation, the regulatory mechanisms and biologic pathways involved are still largely unknown. The hypothesis of this study is that the genesis and growth characteristics of neurofibromatosis type 2 (NF2) associated vestibular schwannomas are determined by genetic alterations that vary in gene transcript expression; this transcript expression includes oncogenic gene products that may be identified by construction and sequencing of a cDNA library from NF2-associated vestibular schwannoma. METHODS: Approximately 3 mL of fresh tumor was obtained during resection of a 4-cm vestibular schwannoma from a patient with NF2. Poly(A)(+) mRNA was isolated, synthesized into double-stranded cDNA, and unidirectionally inserted into Uni-Zap XR (Stratagene, La Jolla, CA) bacteriophage vectors. Bacteriophage vectors containing cDNA inserts were processed into phagemids according to Uni-Zap XR protocol, and inserted vectors were sequenced and analyzed using BLAST software (National Institutes of Health, Bethesda, MD) with GenBank, EMBL, DDBJ, and PBD databases. RESULTS: The cDNA library contained 2.4 million primary plaques. Inserts averaged 1.8 kilobases (kb) in length, with a range of 0.8 to 3.0 kb. BLAST multidatabase comparison of the sequence data obtained from 50 randomly selected clones yielded identification of 13 sequences representing known human genes and 17 sequences representing cloned sequences with unknown function. Three clones represented sequences not previously described in vestibular schwannomas but strongly implicated in oncogenesis within other tissues. CONCLUSIONS: These data have implications for understanding the molecular mechanisms of vestibular schwannoma tumor biology. Identified genes may provide future diagnostic/prognostic markers and therapeutic targets. PMID- 12646840 TI - Dizziness: relating the severity of vertigo to the degree of handicap by measuring vestibular impairment. AB - OBJECTIVE: We sought to correlate the severity of vertigo and handicap in patients with vestibular pathology according to measures of impairment. STUDY DESIGN AND SETTING: We conducted a prospective assessment of patients with dizziness by means of caloric, rotatory test, and computerized dynamic posturography to estimate impairment. Handicap and severity of vertigo were determined with specific questionnaires (Dizziness Handicap Inventory and UCLA DQ). RESULTS: A fair relationship were found between severity of dizziness and vestibular handicap. When impairment was taken into consideration, values were still fair and only moderate for a group of patients with an abnormal caloric test as the only pathologic finding. The composite score from the sensory organization test portion of the computerized dynamic posturography is fairly correlated to severity of vertigo and handicap in the whole population of patients, but no correlation was found when they were assigned to groups of vestibular impairment. CONCLUSION: To assess vestibular impairment, the results from several tests must be taken into account. However, vestibular handicap is not solely explained with measurements of impairment and/or severity. PMID- 12646841 TI - Jugular bulb diverticula: clinical and radiologic aspects. AB - OBJECTIVE: This study was performed to determine the clinical presentation of jugular diverticulum and its relevance with the extension of the lesion. STUDY DESIGN: The records and high-resolution CT scans of 1474 patients, with otologic symptoms or related clinical findings, were evaluated retrospectively. In 17 cases in which CT scans revealed the presence of jugular diverticulum, sensorineural symptoms were evaluated with respect to a reference line, perpendicular to the basal turn of the cochlea and tangent to the vestibule in axial CT scan images. RESULTS: When the extension of jugular diverticulum was posterior to the reference line, sensorineural symptoms with vertigo were dominant in most patients (72.7%) in this group. On the other hand, for the patients with anterior-extending jugular diverticulum, sensorineural symptoms without vertigo were detected in 50% of patients, whereas 33.3% had sensorineural symptoms with vertigo. CONCLUSION: This study revealed a relationship between the extension of jugular diverticulum and clinical symptoms. However, this relationship lacks statistical evidence because of the limited number of patients. PMID- 12646842 TI - Headache after removal of vestibular schwannoma via the retrosigmoid approach: a long-term follow-up-study. AB - OBJECTIVE: Our goal was to study the occurrence and source of origin of postcraniotomy headache syndrome after removal of vestibular schwannoma via the retrosigmoid approach. METHODS: A retrospective chart analysis was conducted of all patients with headache at 3 months after removal of vestibular schwannoma from January 1981 through March 1997 and with a minimum of 24 months of follow up. Diagnosis was made according to the headache classification and was graded using the HARNER scale. Recovery outcome was compared in selected groups of patients with and without headache. A descriptive statistical analysis was used to analyze differences between groups. RESULTS: Of the patients who underwent retrosigmoid craniotomy for removal of vestibular schwannomas, 52 of 155 patients (34%) reported having severe headache of requiring medication every day and/or feeling incapacitated 3 months after surgery. Headache was more prevalent in those who had the bone flap replaced (94% versus 27%), if there was duraplastic or direct dura closure (0% versus 100%). Laboratory-proven aseptic meningitis, most likely due to the use of fibrin glue and drilling of posterior aspect of the internal auditory canal, was mainly associated with postoperative headache (81% versus 2%). In 75% of these cases, calcifications along the brainstem had been noted. CONCLUSION: The origin of postoperative headaches after retrosigmoid vestibular schwannoma resections is not yet fully understood. Different factors may play a role in preventing or reducing headache: dural adhesions to nuchal muscles or to subcutaneous tissues and dural tension in the case of direct dural closure may explain postoperative headache from dural tension. Intradural drilling and the use of fibrin glue may be the source of aseptic meningitis as the etiology of persistent postoperative headache. Prevention of postoperative headache may include the replacement of bone flap at the end of surgery, duraplastic instead of direct dural closure, and prevention of the use of fibrin glue or extensive drilling of the posterior aspect of internal auditory canal. PMID- 12646843 TI - Study of some "intrinsic risk factors" that can enhance an iatrogenic injury of the external branch of the superior laryngeal nerve. AB - OBJECTIVE: This study evaluates some "intrinsic risk factors" for a surgical injury of the external branch of superior laryngeal nerve (EBSLN) based on its anatomic relationship with the thyroid gland. STUDY DESIGN AND SETTING: Seventy two neck dissections were performed in 36 fresh human adult cadavers. Every EBSLN was dissected and categorized using Cernea's classification. All data regarding gender, ethnicity, stature, side of the neck, and volume of hemi thyroid were collected and statistically analyzed by the chi(2)-test and analysis of variance. RESULTS: Although no statistical differences were observed between genders (P = 0.3770), ethnicities (P = 0.9780) or sides of the neck (P = 0.2523), statistical significance was reached regarding the nerve type when the groups were divided by stature (P = 0.0006) and glandular volume (P = 0.0007). CONCLUSION: Individual stature and volume of the hemi thyroid gland are probably involved in risk of the surgical EBSLN injury. SIGNIFICANCE: Surgeons should be aware of these anatomic findings in order to obtain a good voice-related outcome in the thyroidectomies. PMID- 12646844 TI - Viscoelastic properties of rabbit vocal folds after augmentation. AB - BACKGROUND: Vocal fold function is closely related to tissue viscoelasticity. Augmentation substances may alter the viscoelastic properties of vocal fold tissues and hence their vibratory capacity. OBJECTIVES: We sought to investigate the viscoelastic properties of rabbit vocal folds in vitro after injections of various augmentation substances. STUDY DESIGN AND SETTING: Polytetrafluoroethylene (Teflon), cross-linked collagen (Zyplast), and cross linked hyaluronan, hylan b gel (Hylaform) were injected into the lamina propria and the thyroarytenoid muscle of rabbit vocal folds. Dynamic viscosity of the injected vocal fold as a function of frequency was measured with a Bohlin parallel-plate rheometer during small-amplitude oscillation. RESULTS: All injected vocal folds showed a decreasing dynamic viscosity with increasing frequency. Vocal fold samples injected with hylan b gel showed the lowest dynamic viscosity, quite close to noninjected control samples. Vocal folds injected with polytetrafluoroethylene showed the highest dynamic viscosity followed by the collagen samples. CONCLUSIONS: The data indicated that hylan b gel in short-term renders the most natural viscoelastic properties to the vocal fold among the substances tested. This is of importance to restore/preserve the vibratory capacity of the vocal folds when glottal insufficiency is treated with injections. PMID- 12646845 TI - Do micrometastases detected with cytokeratin immunoperoxidase reactivity affect the treatment approach to neck in supraglottic cancers? AB - OBJECTIVES: The aim of this study was to determine the occult lymph node metastasis rates with immunoperoxidase staining for cytokeratin in supraglottic larynx cancers and to assess our approach to the neck. STUDY DESIGN: Twenty-two patients who had squamous cell carcinoma in the supraglottic region and no histopathologic metastasis in the neck who had their cancer detected with the use of hematoxylin-eosin were included in the study. All of the specimens were reevaluated with the use of immunoperoxidase staining for cytokeratin. RESULTS: Micrometastatic disease (pN1) has been detected in 3 patients (13.61%) in whom no metastasis was detected with routine histopathologic examination. CONCLUSION AND SIGNIFICANCE: Results suggest that the occult metastasis rates are higher than those detected with routine hematoxylin-eosin staining. However, because it is accepted that neck dissection alone has adequate therapeutic efficiency in pN1 patients, the detected 13.61% rate of micrometastases has not changed our treatment plan in the neck. PMID- 12646846 TI - Functional comparison after reconstruction with a radial forearm free flap or a pectoralis major flap for cancer of the tongue. AB - OBJECTIVE: Numerous patients in Taiwan with tongue carcinoma require tongue reconstruction. We compared the abilities of 2 methods of tongue reconstruction to reserve tongue function. STUDY DESIGN AND SETTING: Sixty patients underwent resection of the tumors and reconstruction with a pectoralis major flap or a radial forearm flap. The Chinese articulation test was used to evaluate the place and manner of error production, and a questionnaire on dietary habits was used to evaluate deglutition 6 months to 10 years after reconstruction. RESULTS: Patients with the free flap had more intelligible speech. The questionnaire study showed no significant difference between the 2 groups in swallowing rating. Motility caused by flap pliability increased speech intelligibility more than it did on swallowing function. CONCLUSION: Our experience in a few selected patients shows that the functional outcome of tongue surgery is related to the reconstruction methods used (for speech) and to the extent of tongue resection (for swallowing). PMID- 12646847 TI - Multiple primary tumors in patients diagnosed with hypopharyngeal cancer. AB - OBJECTIVE: There have been few series to report on the incidence of multiple primary tumors associated with hypopharyngeal cancer. A unique consecutive patient group in a closed community who were treated by a single surgeon was available. The incidence and effect of multiple primary tumors were unknown. STUDY DESIGN: We sought to assess (1) the incidence of multiple primary tumors among patients with hypopharyngeal cancer who were treated at a tertiary center, (2) the incidence of synchronous and metachronous tumors, and (3) the location of these multiple primary tumors and their effect on patient survival. METHODS: We conducted a retrospective study of case notes of 150 consecutive patients with hypopharyngeal malignancy treated by a single surgeon between 1983 and 1998. Information was compiled from the patients' medical records and death data from the Family Health Services Authority. RESULTS: Thirty-four patients had multiple primary tumors (22.6%). There were 22 men and 12 women; piriform fossa tumor was seen in 21 men and 6 women, and postcricoid space tumor was seen in 6 women and 1 man. Second primary tumors were synchronous in 7 patients, subsequent to hypopharyngeal tumor in 5 patients, and antecedent to hypopharyngeal tumor in 14 patients. Eight patients had 2 primary tumors, of which 4 were synchronous, 4 were subsequent, and 8 were antecedent to hypopharyngeal malignancy. On the last review (2001), 3 patients were alive, and 31 had died: 17 had died from primary malignancy, 11 from another malignancy, and 3 from unrelated causes. CONCLUSION: The presence of second primary tumors in hypopharyngeal cancer is higher than previously reported, and their presence had a significant effect on the patients' survival. PMID- 12646848 TI - Maxillary infraorbital myxoma: reconstruction with vascularized temporal bone. PMID- 12646849 TI - Lingual dermoid cysts. PMID- 12646850 TI - Hemangioma of the left cheek: a case report. PMID- 12646851 TI - Protrusion of orbital content through dehiscence of lamina papyracea mimics ethmoiditis: a case report. PMID- 12646852 TI - Zygomatic abscess complicating a huge mastoid cholesteatoma with intact eardrum. PMID- 12646853 TI - Eccrine poroma in the external auditory canal. PMID- 12646854 TI - Bilateral myoclonus of the tensor tympani: a case report. PMID- 12646855 TI - Leiomyosarcoma of the auricle. PMID- 12646856 TI - Angiocentric eosinophilic fibrosis. PMID- 12646858 TI - A novel attenuated replication-competent adenovirus for melanoma therapy. AB - To generate a replication-competent adenovirus (Ad) with specificity for melanoma, we constructed a tissue-specific promoter restricting E1A expression to melanoma cells. The combination of four copies of a mouse tyrosinase enhancer element (TE) fused to the human tyrosinase promoter (TP) yielded up to 2000-fold higher luciferase reporter activity in tyrosinase-expressing melanoma cells than in nonmelanoma cells. Insertion of the composite TETP construct upstream of the E1A gene was combined with deleting as far as possible the intertwined endogenous Ad enhancer/promoter (EP). The resulting AdDeltaEP-TETP vector, also deleted for the E3 region, was found to replicate in tyrosinase-positive melanoma cells, such as SK-Mel23 as efficiently as wild-type Ad5, but at a more than 50-fold reduced level in nonmelanoma tumour cells and primary human cells. Injection of AdDeltaEP TETP into xenotransplanted melanomas, but not into HeLa-derived tumours led to long-lasting tumour regression in nude mice. This AdDeltaEP-TETP virus might be useful for the treatment of accessible lesions in advanced melanoma patients. PMID- 12646860 TI - Amelioration of antigen-induced arthritis in rats by transfer of extracellular superoxide dismutase and catalase genes. AB - Reactive oxygen species (ROS) have been implicated in the pathogenesis of rheumatoid arthritis (RA), while antioxidant enzymes, such as extracellular superoxide dismutase (EC-SOD) and catalase, block radical-induced events. The present study tested if the ex vivo transfer of EC-SOD and catalase genes alone or in combination in the knee joint of rats with monoarticular antigen-induced arthritis (AIA) was anti-inflammatory, and examined the potential mechanisms involved. Synoviocytes isolated from female Wistar rats were immortalized with a retroviral vector SUV19.5. These cells were permanently transfected with an EC SOD expression plasmid (pEC-SODZeo) or a catalase expression plasmid (pCatalaseZeo) to create cells overexpressing EC-SOD or catalase, as measured by RT-PCR and Western blots. The cells were engrafted in knee joints of animals at the time of the induction of AIA. Three gene transfer groups, an EC-SOD group, a catalase group and a combined therapy group (EC-SOD and catalase) were included in these experiments. Animals in the control group were engrafted with synoviocytes transfected with the plasmid pZeoSV2 without an insert. Clinical and histological assessments were performed, as well as tissue measurements of SOD, catalase and gelatinase activities. Ex vivo gene transfer of EC-SOD and catalase into rat knee joints produced about a six- to seven-fold increase in EC-SOD activity and a two- to three-fold increase in catalase activity compared with the control animals. Rats treated with cells overexpressing EC-SOD, catalase or a combination of EC-SOD and catalase showed significant suppression of knee joint swelling, decreased infiltration of inflammatory cells within the synovial membrane and reduced gelatinase activity in knee joints, compared with animals receiving cells transfected with the plasmid alone. No statistically significant difference was found between the groups treated with cells overexpressing EC-SOD, catalase or a combination of both. Gene therapy involving the local intra articular overexpression of two antioxidant enzymes, EC-SOD and catalase, was anti-inflammatory in AIA. One mechanism appears to be the suppression of gelatinase activities by both EC-SOD and catalase. PMID- 12646859 TI - Hypoxia-mediated tumour targeting. AB - Hypoxia is a common physiological feature of tumours. It activates a signalling cascade that culminates in the stabilization of the HIF-1 transcription factor and activation of genes that possess a hypoxia response element (HRE). We have used an optimized hypoxia responsive promoter (OBHRE) to investigate hypoxia targeted gene expression in vivo in the context of an adenovirus vector. The OBHRE promoter showed limited activity in the liver or spleen such that expression was 1000-fold lower than that driven by the strong CMV/IE promoter. However, in the context of the tumour microenvironment, the OBHRE promoter achieved expression levels comparable to that of the CMV/IE promoter. Next, we showed that an adenovirus expressing the human cytochrome P450 (CYP2B6) regulated by the OBHRE promoter delays tumour growth in response to the prodrug cyclophosphamide (CPA). Finally, we exploited the hepatotropism of adenovirus to investigate whether the OBHRE promoter could mitigate the hepatotoxicity of a recombinant adenovirus expressing thymidine kinase (TK) in the context of the prodrug ganciclovir (GCV). High-dose Ad.CMVTK/GCV treatment caused significant liver necrosis whereas the same dose of Ad.HRETK was well tolerated. These in vivo data demonstrate that hypoxia-targeted gene expression via the OBHRE promoter can be used to increase the therapeutic window of cytotoxic cancer gene therapy. PMID- 12646861 TI - Suppression of glomerulosclerosis by adenovirus-mediated IL-10 expression in the kidney. AB - Glomerulosclerosis is a common morphologic result seen in almost all progressed renal diseases, and is the characteristic change in focal segmental glomerulosclerosis (FSGS). The most convincing hypothesis for glomerulosclerosis is cytokine-mediated injury by infiltrating immune cells in the glomerulus and tubulointerstitial area. This study investigated whether the anti-inflammatory effect of interleukin-10 (IL-10) when expressed by a recombinant adenoviral vector can prevent the onset of glomerulosclerosis in FGS/Kist mice (an animal model with naturally occurring renal failure initiated by FSGS). Each group of mice received recombinant adenoviruses encoding human IL-10 (Ad:hIL-10) by intraparenchymal injection at 6 weeks and were examined for cytokine expression, glomerular sclerotic index, and proteinuria. After injection of Ad:hIL-10 to the kidney, IL-10 expression was found to last over 20 days. Mice treated with Ad:hIL 10 were shown to have a significant reduction in the glomerular sclerotic index at 10 weeks when compared to control groups. The level of proteinuria in Ad:hIL 10-treated mice was also significantly reduced. About 50% of the urine samples of naive and Ad:LacZ-treated groups had severe levels of proteinuria. By contrast, at 10 weeks the group treated with Ad:hIL-10 had lower levels of proteinuria and transforming growth factor-beta1 (TGF-beta1) expression. These results demonstrate that IL-10 effectively prevents the development of glomerulosclerosis in FGS/Kist mice, and IL-10 gene therapy may be of use for the treatment of renal failure. PMID- 12646863 TI - Transfection of oocytes and other types of ovarian cells in rabbits after direct injection into uterine arteries of adenoviruses and plasmid/liposomes. AB - Transfection of oocytes should be avoided in somatic gene therapy. However, several viral vectors including adenoviruses can transfect zona-pellucida-free eggs in vitro. During early stages of development, oocytes of postnatal ovaries lack the zona pellucida. Therefore, they may be susceptible to gene transfer and unintended toxic effects. The purpose of this study was to see whether the injection of adenoviruses (1 x 10(10) PFU) or plasmid (500 microg)/DOTMA:DOPE (1:2) liposomes directly into uterine arteries in pregnant rabbits leads to transfection of oocytes and other types of ovarian cells. LacZ and herpes simplex virus thymidine kinase (HSV-TK) were used as transgenes. It was found that both adenovirus and plasmid vectors transfected oocytes at the primordial and primary follicle stage when they were not protected by the zona pellucida, whereas no transfection was seen in oocytes surrounded by the zona pellucida. Efficient transfection of corpus luteum and granulosa cells was also detected by adenoviral and plasmid vectors. Transfection of oocytes and other ovarian cells was verified by X-gal staining and laser microdissection, followed by PCR analysis. HSV-TK gene transfer, followed by ganciclovir treatment, led to destruction of a significant number of oocytes, whereas HSV-TK gene transfer alone did not lead to toxic effects. It is concluded that the presence of a high concentration of adenovirus or plasmid vectors via the uterine artery may lead to transfection of zona-pellucida-free oocytes and other ovarian cells. PMID- 12646862 TI - Non-viral gene transfer of murine spleen cells achieved by in vivo electroporation. AB - Gene electrotranfer is an attractive physical method to deliver genes to target tissues. The aim of this study was to evaluate in vivo gene electrotransfer into spleen, one of the most important lymphoid organ, in order to create a new tool to modulate the immuno-inflammatory system. C57Bl/6 mice were submitted either to intramuscular electrotransfer (IME) as a reference method or to intrasplenic (ISE) gene electrotransfer. In the naked injected plasmids, the CMV promoter controlled the expression of luciferase, secreted alkaline phosphatase, EGFP, or IFNgamma. The ISE optimal electrotransfer conditions were first determined and ISE was found to be an efficient gene transfer method, which can be used to express secreted or intracellular proteins transiently. Although transfected cells were still present in the spleen 30 days after ISE, transfected spleen cells could recirculate since they were detected in extrasplenic locations. Using a T-lymphocyte-specific promoter controlling the expression of EGFP, splenic T cells could be targeted. Finally, it appeared that ISE procedure does not impair by itself the immune response and does not result in a significant production of antibodies directed to the transgenic proteins in C57Bl/6 mice. This strategy constitutes a new method to manipulate the immune response that can be used in various experimental designs. PMID- 12646864 TI - Water-soluble lipopolymer as an efficient carrier for gene delivery to myocardium. AB - Water-soluble lipopolymer (WSLP), which consisted of polyethylenimine (PEI, 1800 Da) and cholesterol, was characterized as a gene carrier to smooth muscle cells and myocardium. Acid-base titration showed that WSLP had a proton-buffering effect. The size of WSLP/plasmid DNA (pDNA) complex was around 70 nm. WSLP/pDNA complex was transfected to A7R5 cells, a smooth muscle cell line. WSLP showed the highest transfection at a 40/1 N/P ratio. WSLP has higher transfection efficiency than PEI (1800 and 25 000 Da), SuperFect, and lipofectamine. In addition, WSLP has less cytotoxicity than PEI (25 000 Da), SuperFect, and lipofectamine. Since WSLP has cholesterol moiety, it may utilize cellular cholesterol uptake pathway, in which low-density lipoprotein (LDL) is involved. An inhibition study with free cholesterol or low-density lipoprotein (LDL) showed that transfection was inhibited by cholesterol or LDL, suggesting that WSLP/pDNA complex is transfected to the cells through the cholesterol uptake pathway. To evaluate the transfection efficiency to myocardium, WSLP/pDNA complex was injected into the rabbit myocardium. WSLP showed higher transfection than PEI and naked pDNA. WSLP expressed the transgene for more than 2 weeks. In conclusion, WSLP is an efficient carrier for local gene transfection to myocardium, and useful in in vivo gene therapy. PMID- 12646865 TI - Antigen-specific targeting of CD8+ T cells with receptor-modified T lymphocytes. AB - Chimeric receptors that link ligand recognition domains, such as antibody Fv fragments, with TCR signaling domains can redirect T lymphocytes against MHC unrestricted targets. Such receptor-modified T lymphocytes have shown promise in the treatment of infectious diseases and cancer. We hypothesized that receptor modified T lymphocytes may also be designed to target antigen-specific T cells. We synthesized chimeric receptors consisting of the extracellular and transmembrane domains of the class I MHC H-2K(b) molecule linked to the signaling domains of either TCR-zeta, CD28 and zeta, or CD28, zeta, and lck. T lymphocytes modified to express these receptors and pulsed with antigenic peptide specifically killed precursor CTL. Cytolysis was efficient, even at effector:target ratios of less than one, and specific, selectively killing antigen-specific precursor CTL among a mixed population of T cells. Cytolysis required activation of the receptor-modified T cells, and did not occur with a signaling-deficient chimeric receptor. In contrast to precursor CTL, differentiated CTL proved resistant to lysis by the receptor-modified T cells. These data demonstrate the feasibility of redirecting T lymphocytes against antigen-specific T cells. Receptor-modified T cells expressing chimeric MHC receptors have potential application in autoimmune and alloimmune diseases. PMID- 12646867 TI - Dynamic proteins and a cytoskeleton in bacteria. PMID- 12646868 TI - Armadillo/beta-catenin signals in the nucleus--proof beyond a reasonable doubt? AB - Wnt signalling results in transcriptional stimulation of genes controlling normal and malignant development. A key effector of the canonical Wnt pathway is beta catenin (also known as Drosophila melanogaster Armadillo (Arm)), thought to function as a nuclear co-activator of TCF transcription factors. This has been challenged by unexpected observations of membrane-bound Arm/beta-catenin signalling activity. Plausible explanations allow these observations to be reconciled with the large body of evidence supporting a nuclear function of Arm/beta-catenin. PMID- 12646869 TI - A nucleolar disappearing act in somatic cloning. PMID- 12646870 TI - Rac signalling: a radical view. PMID- 12646871 TI - RNA repaired. PMID- 12646872 TI - Multitasking at mitotic exit. PMID- 12646873 TI - GNOM alone. PMID- 12646874 TI - The multiple functions of tumour suppressors: it's all in APC. PMID- 12646879 TI - How to slake a planet's thirst. PMID- 12646877 TI - The spindle-associated transmembrane protein Axs identifies a membranous structure ensheathing the meiotic spindle. AB - Mutations in the aberrant X segragation (Axs) gene disrupt the segregation of achiasmate chromosomes during female meiosis in Drosophila melanogaster. We show that Axs encodes the founding member of an eukaryotic family of transmembrane proteins. Axs protein colocalizes with components of the endoplasmic reticulum and is present within a structure ensheathing the meiotic spindle. In both meiotic and mitotic cells, Axs is recruited to the microtubules of assembling spindles. We propose that Axs and the sheath represent novel mediators of meiotic spindle assembly and chromosome segregation. PMID- 12646880 TI - Biodefence plans earn lukewarm response from US academics. PMID- 12646881 TI - Biotech firms join charities in drive to help Africa's farms. PMID- 12646882 TI - Health labs focus on mystery pneumonia. PMID- 12646884 TI - Revamped accelerator seeks quark's secrets. PMID- 12646883 TI - Chicken flu races through Dutch poultry farms. PMID- 12646885 TI - NATO reform promises more publicity for less science. PMID- 12646886 TI - Alien-hunters get scope to search. PMID- 12646887 TI - NASA intensifies probe into Columbia failure. PMID- 12646889 TI - The world's forgotten crisis. PMID- 12646890 TI - Atlas of a thirsty planet. PMID- 12646891 TI - Delta blues. PMID- 12646892 TI - If atomic precision is unfeasible, so is life. PMID- 12646893 TI - Venezuela: crisis puts major institutions at risk. PMID- 12646894 TI - Venezuela: the other side of the story. PMID- 12646895 TI - The politics of publication. PMID- 12646903 TI - Cell biology: Just mix and patch. PMID- 12646904 TI - Astronomy: Glowing embers. PMID- 12646905 TI - Genomics: Gene expression meets genetics. PMID- 12646906 TI - Optics: Ado about nothing much? PMID- 12646909 TI - Obituary: Jean Brossel (1918-2003). PMID- 12646907 TI - Developmental biology: A hairy situation. PMID- 12646908 TI - Meteorology: Getting the wind up. PMID- 12646911 TI - Surveillance: Radiographic imaging with cosmic-ray muons. PMID- 12646912 TI - Palaeo-oceanography: Deepwater variability in the Holocene epoch. PMID- 12646913 TI - Reduced drag coefficient for high wind speeds in tropical cyclones. AB - The transfer of momentum between the atmosphere and the ocean is described in terms of the variation of wind speed with height and a drag coefficient that increases with sea surface roughness and wind speed. But direct measurements have only been available for weak winds; momentum transfer under extreme wind conditions has therefore been extrapolated from these field measurements. Global Positioning System sondes have been used since 1997 to measure the profiles of the strong winds in the marine boundary layer associated with tropical cyclones. Here we present an analysis of these data, which show a logarithmic increase in mean wind speed with height in the lowest 200 m, maximum wind speed at 500 m and a gradual weakening up to a height of 3 km. By determining surface stress, roughness length and neutral stability drag coefficient, we find that surface momentum flux levels off as the wind speeds increase above hurricane force. This behaviour is contrary to surface flux parameterizations that are currently used in a variety of modelling applications, including hurricane risk assessment and prediction of storm motion, intensity, waves and storm surges. PMID- 12646914 TI - Early optical emission from the gamma-ray burst of 4 October 2002. AB - Observations of the long-lived emission--or 'afterglow'--of long-duration gamma ray bursts place them at cosmological distances, but the origin of these energetic explosions remains a mystery. Observations of optical emission contemporaneous with the burst of gamma-rays should provide insight into the details of the explosion, as well as into the structure of the surrounding environment. One bright optical flash was detected during a burst, but other efforts have produced negative results. Here we report the discovery of the optical counterpart of GRB021004 only 193 seconds after the event. The initial decline is unexpectedly slow and requires varying energy content in the gamma-ray burst blastwave over the course of the first hour. Further analysis of the X-ray and optical afterglow suggests additional energy variations over the first few days. PMID- 12646915 TI - Femtosecond X-ray measurement of coherent lattice vibrations near the Lindemann stability limit. AB - The study of phase-transition dynamics in solids beyond a time-averaged kinetic description requires direct measurement of the changes in the atomic configuration along the physical pathways leading to the new phase. The timescale of interest is in the range 10(-14) to 10(-12) s. Until recently, only optical techniques were capable of providing adequate time resolution, albeit with indirect sensitivity to structural arrangement. Ultrafast laser-induced changes of long-range order have recently been directly established for some materials using time-resolved X-ray diffraction. However, the measurement of the atomic displacements within the unit cell, as well as their relationship with the stability limit of a structural phase, has to date remained obscure. Here we report time-resolved X-ray diffraction measurements of the coherent atomic displacement of the lattice atoms in photoexcited bismuth close to a phase transition. Excitation of large-amplitude coherent optical phonons gives rise to a periodic modulation of the X-ray diffraction efficiency. Stronger excitation corresponding to atomic displacements exceeding 10 per cent of the nearest neighbour distance-near the Lindemann limit-leads to a subsequent loss of long range order, which is most probably due to melting of the material. PMID- 12646916 TI - Phonon interpretation of the 'boson peak' in supercooled liquids. AB - Glasses are amorphous solids, in the sense that they display elastic behaviour. In crystalline solids, elasticity is associated with phonons, which are quantized vibrational excitations. Phonon-like excitations also exist in glasses at very high (terahertz; 10(12) Hz) frequencies; surprisingly, these persist in the supercooled liquids. A universal feature of such amorphous systems is the boson peak: the vibrational density of states has an excess compared to the Debye squared-frequency law. Here we investigate the origin of this feature by studying the spectra of inherent structures (local minima of the potential energy) in a realistic glass model. We claim that the peak is the signature of a phase transition in the space of the stationary points of the energy, from a minima dominated phase (with phonons) at low energy to a saddle-point-dominated phase (without phonons). The boson peak moves to lower frequencies on approaching the phonon-saddle transition, and its height diverges at the critical point. Our numerical results agree with the predictions of euclidean random matrix theory on the existence of a sharp phase transition between an amorphous elastic phase and a phonon-free one. PMID- 12646917 TI - Detection of human influence on sea-level pressure. AB - Greenhouse gases and tropospheric sulphate aerosols--the main human influences on climate--have been shown to have had a detectable effect on surface air temperature, the temperature of the free troposphere and stratosphere and ocean temperature. Nevertheless, the question remains as to whether human influence is detectable in any variable other than temperature. Here we detect an influence of anthropogenic greenhouse gases and sulphate aerosols in observations of winter sea-level pressure (December to February), using combined simulations from four climate models. We find increases in sea-level pressure over the subtropical North Atlantic Ocean, southern Europe and North Africa, and decreases in the polar regions and the North Pacific Ocean, in response to human influence. Our analysis also indicates that the climate models substantially underestimate the magnitude of the sea-level pressure response. This discrepancy suggests that the upward trend in the North Atlantic Oscillation index (corresponding to strengthened westerlies in the North Atlantic region), as simulated in a number of global warming scenarios, may be too small, leading to an underestimation of the impacts of anthropogenic climate change on European climate. PMID- 12646918 TI - Enhanced mantle-to-crust rhenium transfer in undegassed arc magmas. AB - Variations in the 187Os/188Os isotopic signature of mantle and mantle-derived rocks have been thought to provide a powerful chemical tracer of deep Earth structure. Many studies have inferred from such data that a long-lived, high rhenium component exists in the deep mantle (187Re is the parent isotope decaying to 187Os, with a half-life of approximately 42 billion years), and that this reservoir probably consists of subducted oceanic crust. The interpretation of these isotopic signatures is, however, dependent on accurate estimates of rhenium and osmium concentrations in all of the main geochemical reservoirs, and the crust has generally been considered to be a minor contributor to such global budgets. In contrast, we here present observations of high rhenium concentrations and low Yb/Re ratios in arc-type melt inclusions. These results indicate strong enrichment of rhenium in undegassed arc rocks, and consequently the continental crust, which results in a crustal estimate of 2 p.p.b. rhenium, as compared to previous estimates of 0.4-0.2 p.p.b. (refs 4, 5). Previous determinations of rhenium in arc materials, which were largely measured on subaerially erupted samples, are likely to be in error owing to rhenium loss during degassing. High mantle-to-crust rhenium fluxes, as observed here, require a revaluation of geochemical models based on the 187Re-187Os decay system. PMID- 12646919 TI - Genetics of gene expression surveyed in maize, mouse and man. AB - Treating messenger RNA transcript abundances as quantitative traits and mapping gene expression quantitative trait loci for these traits has been pursued in gene specific ways. Transcript abundances often serve as a surrogate for classical quantitative traits in that the levels of expression are significantly correlated with the classical traits across members of a segregating population. The correlation structure between transcript abundances and classical traits has been used to identify susceptibility loci for complex diseases such as diabetes and allergic asthma. One study recently completed the first comprehensive dissection of transcriptional regulation in budding yeast, giving a detailed glimpse of a genome-wide survey of the genetics of gene expression. Unlike classical quantitative traits, which often represent gross clinical measurements that may be far removed from the biological processes giving rise to them, the genetic linkages associated with transcript abundance affords a closer look at cellular biochemical processes. Here we describe comprehensive genetic screens of mouse, plant and human transcriptomes by considering gene expression values as quantitative traits. We identify a gene expression pattern strongly associated with obesity in a murine cross, and observe two distinct obesity subtypes. Furthermore, we find that these obesity subtypes are under the control of different loci. PMID- 12646920 TI - Glycine binding primes NMDA receptor internalization. AB - NMDA (N-methyl-d-aspartate) receptors (NMDARs) are a principal subtype of excitatory ligand-gated ion channel with prominent roles in physiological and disease processes in the central nervous system. Recognition that glycine potentiates NMDAR-mediated currents as well as being a requisite co-agonist of the NMDAR subtype of 'glutamate' receptor profoundly changed our understanding of chemical synaptic communication in the central nervous system. The binding of both glycine and glutamate is necessary to cause opening of the NMDAR conductance pore. Although binding of either agonist alone is insufficient to cause current flow through the channel, we report here that stimulation of the glycine site initiates signalling through the NMDAR complex, priming the receptors for clathrin-dependent endocytosis. Glycine binding alone does not cause the receptor to be endocytosed; this requires both glycine and glutamate site activation of NMDARs. The priming effect of glycine is mimicked by the NMDAR glycine site agonist d-serine, and is blocked by competitive glycine site antagonists. Synaptic as well as extrasynaptic NMDARs are primed for internalization by glycine site stimulation. Our results demonstrate transmembrane signal transduction through activating the glycine site of NMDARs, and elucidate a model for modulating cell-cell communication in the central nervous system. PMID- 12646921 TI - Antibody neutralization and escape by HIV-1. AB - Neutralizing antibodies (Nab) are a principal component of an effective human immune response to many pathogens, yet their role in HIV-1 infection is unclear. To gain a better understanding of this role, we examined plasma from patients with acute HIV infection. Here we report the detection of autologous Nab as early as 52 days after detection of HIV-specific antibodies. The viral inhibitory activity of Nab resulted in complete replacement of neutralization-sensitive virus by successive populations of resistant virus. Escape virus contained mutations in the env gene that were unexpectedly sparse, did not map generally to known neutralization epitopes, and involved primarily changes in N-linked glycosylation. This pattern of escape, and the exceptional density of HIV-1 envelope glycosylation generally, led us to postulate an evolving 'glycan shield' mechanism of neutralization escape whereby selected changes in glycan packing prevent Nab binding but not receptor binding. Direct support for this model was obtained by mutational substitution showing that Nab-selected alterations in glycosylation conferred escape from both autologous antibody and epitope-specific monoclonal antibodies. The evolving glycan shield thus represents a new mechanism contributing to HIV-1 persistence in the face of an evolving antibody repertoire. PMID- 12646922 TI - Links between signal transduction, transcription and adhesion in epithelial bud development. AB - The morphogenesis of organs as diverse as lungs, teeth and hair follicles is initiated by a downgrowth from a layer of epithelial stem cells. During follicular morphogenesis, stem cells form this bud structure by changing their polarity and cell-cell contacts. Here we show that this process is achieved through simultaneous receipt of two external signals: a Wnt protein to stabilize beta-catenin, and a bone morphogenetic protein (BMP) inhibitor to produce Lef1. Beta-catenin then binds to, and activates, Lef1 transcription complexes that appear to act uncharacteristically by downregulating the gene encoding E cadherin, an important component of polarity and intercellular adhesion. When either signal is missing, functional Lef1 complexes are not made, and E-cadherin downregulation and follicle morphogenesis are impaired. In Drosophila, E-cadherin can influence the plane of cell division and cytoskeletal dynamics. Consistent with this notion, we show that forced elevation of E-cadherin levels block invagination and follicle production. Our findings reveal an intricate molecular programme that links two extracellular signalling pathways to the formation of a nuclear transcription factor that acts on target genes to remodel cellular junctions and permit follicle formation. PMID- 12646924 TI - Insights into the ubiquitin transfer cascade from the structure of the activating enzyme for NEDD8. AB - Post-translational modification by ubiquitin-like proteins (Ublps) is an essential cellular regulatory mechanism. The Ublp NEDD8 regulates cell division, signalling and embryogenesis. Ublps are conjugated to their targets by the sequential action of E1, E2 and often E3 enzymes. Each Ublp has a dedicated E1, or activating enzyme, that initiates its conjugation cascade. First, E1 associates with the Ublp and catalyses adenylation of the carboxy terminus of the Ublp. Second, E1 forms a thioester between its catalytic cysteine and the Ublp. Next, E1 is loaded with a second Ublp molecule, adenylating the C terminus of this second Ublp while still carrying the first thioester-bound Ublp. Last, E1 binds E2 and promotes Ublp transfer to the catalytic cysteine of E2. We report here the structure and mutational analysis of human APPBP1-UBA3, the heterodimeric E1 enzyme for NEDD8 (ref. 11). Each E1 activity is specified by a domain: an adenylation domain resembling bacterial adenylating enzymes, an E1 specific domain organized around the catalytic cysteine, and a domain involved in E2 recognition resembling ubiquitin. The domains are arranged around two clefts that coordinate protein and nucleotide binding so that each of E1's reactions drives the next, in an assembly-line fashion. PMID- 12646926 TI - A question of form. PMID- 12646923 TI - Segregation of receptor and ligand regulates activation of epithelial growth factor receptor. AB - Interactions between ligands and receptors are central to communication between cells and tissues. Human airway epithelia constitutively produce both a ligand, the growth factor heregulin, and its receptors--erbB2, erbB3 and erbB4 (refs 1 3). Although heregulin binding initiates cellular proliferation and differentiation, airway epithelia have a low rate of cell division. This raises the question of how ligand-receptor interactions are controlled in epithelia. Here we show that in differentiated human airway epithelia, heregulin-alpha is present exclusively in the apical membrane and the overlying airway surface liquid, physically separated from erbB2-4, which segregate to the basolateral membrane. This physical arrangement creates a ligand-receptor pair poised for activation whenever epithelial integrity is disrupted. Indeed, immediately following a mechanical injury, heregulin-alpha activates erbB2 in cells at the edge of the wound, and this process hastens restoration of epithelial integrity. Likewise, when epithelial cells are not separated into apical and basolateral membranes ('polarized'), or when tight junctions between adjacent cells are opened, heregulin-alpha activates its receptor. This mechanism of ligand-receptor segregation on either side of epithelial tight junctions may be vital for rapid restoration of integrity following injury, and hence critical for survival. This model also suggests a mechanism for abnormal receptor activation in diseases with increased epithelial permeability. PMID- 12646927 TI - Drug target validation: Hitting the target. PMID- 12646928 TI - The silent treatment. PMID- 12646929 TI - A whole picture. PMID- 12646931 TI - The glory of autonomy. PMID- 12646932 TI - Victims of success. PMID- 12646933 TI - Clouded vision. PMID- 12646934 TI - Demonstration of Philadelphia chromosome negative abnormal clones in patients with chronic myelogenous leukemia during major cytogenetic responses induced by imatinib mesylate. AB - Imatinib mesylate, an Abl-specific kinase inhibitor, produces sustained complete hematologic responses (CHR) and major cytogenetic responses (MCR) in chronic myeloid leukemia (CML) patients, but long-term outcomes in these patients are not yet known. This article reports the identification of clonal abnormalities in cells lacking detectable Philadelphia (Ph) chromosome/BCR-ABL rearrangements from seven patients with chronic- or accelerated-phase CML, who were treated with imatinib. All seven patients were refractory or intolerant to interferon therapy. Six of seven patients demonstrated MCR and one patient, who had a cryptic translocation, achieved low-level positivity (2.5%) for BCR-ABL by fluorescence in situ hybridization. The median duration of imatinib treatment before the identification of cytogenetic abnormalities in BCR-ABL-negative cells was 13 months. The most common cytogenetic abnormality was trisomy 8, documented in three patients. All patients had varying degrees of dysplastic morphologic abnormalities. One patient exhibited increased numbers of marrow blasts, yet consistently demonstrated no Ph-positive metaphases and the absence of morphologic features of CML. The presence of clonal abnormalities in Ph-negative cells of imatinib-treated CML patients with MCR and CHR highlights the importance of routine metaphase cytogenetic testing and long-term follow-up of all imatinib treated patients. PMID- 12646935 TI - Is modulation of multidrug resistance a viable strategy for acute myeloid leukemia? PMID- 12646936 TI - New targets for therapy in acute myeloid leukemia. PMID- 12646937 TI - Induction and postremission therapy: new agents. PMID- 12646938 TI - Elitek-rasburicase: an effective means to prevent and treat hyperuricemia associated with tumor lysis syndrome, a Meeting Report, Dallas, Texas, January 2002. AB - Renal precipitation of uric acid associated with tumor lysis syndrome (TLS) is a major complication in the management of leukemia, lymphoma, and other drug sensitive cancers. Management of hyperuricema has historically consisted of administration of allopurinol, hydration, alkalinization to maintain pH between 7.0 and 7.3, and in some cases diuresis. Allopurinol, a xanthine analogue, blocks xanthine oxidase and formation of uric acid. Urate oxidase converts uric acid to allantoin, which is 5-10 times more soluble than uric acid. Homo sapiens cannot express urate oxidase because of a nonsense mutation. Urate oxidase was initially purified from Aspergillus flavus fungus. Treatment with this nonrecombinant product had been effective in preventing renal precipitation of uric acid in cancer patients, but was associated with a relatively high frequency of allergic reactions. This enzyme was recently cloned from A. flavus and is now manufactured as a recombinant protein. Clinical trials have shown this drug to be more effective than allopurinol for prevention and treatment of hyperuricemia in leukemia and lymphoma patients. This drug has been approved in Europe as well as the US and several clinical trials are in progress to further determine its clinical utility in other patient subsets. The purpose of this meeting was to discuss usefulness of recombinant urate oxidase, also known as rasburicase, Fasturtec, and Elitek, for the management of TLS in certain cancer patients. PMID- 12646939 TI - Immunological classification of acute myeloblastic leukemias: relevance to patient outcome. AB - Immunophenotyping is a major tool to assign acute leukemia blast cells to the myeloid lineage. However, because of the large heterogeneity of myeloid-related lineages, no clinically relevant immunological classification of acute myeloblastic leukemia (AML) has been devised so far. To attempt at formulating such a classification, we analyzed the pattern of expression of selected antigens, on blast cells collected at AML diagnosis. Patients were eligible if they had a first diagnosis of de novo AML and a sufficient number of blast cells for proper immunophenotyping. The relative expression of CD7, CD13, CD14, CD15, CD33, CD34, CD35, CD36, CD65, CD117, and HLA-DR were analyzed by cytometry in a test series of 176 consecutive AML cases. Statistical tools of clusterization allowed to remove antigens with overlapping distribution, leading us to propose an AML classification that was validated in a second AML cohort of 733 patients. We identified five AML subsets (MA to ME) based on the expression of seven antigens within four groups (CD13/CD33/CD117, CD7, CD35/CD36, CD15).-MA and MB AML have exclusively myeloid features with seldom extramedullary disease and rare expression of lymphoid antigens. No cases of acute promyelocytic leukemia (APL) were observed within MB AML. MC AML have either myeloid or erythroblastic features. MD AML have more frequently high WBC counts than other subsets, which were related to the expression of CD35/CD36 and CD14 and to monoblastic differentiation. ME AML lack CD13, CD33, and CD117 but display signs of terminal myeloid differentiation. Specific independent prognostic factors were related to poor overall survival in each immunological subset: CD34+ (P<3 x 10(-4)) in MA AML, CD7+ in MB AML, non-APL cases (P<0.03) in MC AML, CD34+ (P<0.002) and CD14+ (P<0.03) in MD AML, CD14+ in ME AML (P<0.01). The inclusion of seven key markers in the immunophenotyping of AML allows a stratification into clinically relevant subsets with individual prognostic factors, which should be considered to define high-risk AML populations. PMID- 12646940 TI - Deletions on der(9) chromosome in adult Ph-positive acute lymphoblastic leukemia occur with a frequency similar to that observed in chronic myeloid leukemia. AB - The t(9;22)(q34;q11), generating the Philadelphia chromosome (Ph), is found in more than 90% of patients with chronic myeloid leukemia (CML) and in 15-30% of adults with acute lymphoblastic leukemia (ALL). Different groups have recently described the presence of large genomic deletions adjacent to the translocation breakpoint on the derivative chromosome 9 in 9-16% of CML patients. In the present paper, we report a FISH study of 45 Ph+ adult ALL patients with the aim of investigating the presence of deletions on derivative chromosome 9. In four (9%) of 45 cases, all showing an M-bcr, we detected deletions on der(9). The frequency of deletions we observed is similar to that reported in CML patients. The association of an M-bcr breakpoint and deletions appears significant (P=0.03). Some authors have suggested a very low incidence of der(9) deletions in ALL. This discrepancy can be explained by taking into account the low percentage of M-bcr ALL patients in the latter study (18%) compared to the present one (44%). PMID- 12646941 TI - Modification of topoisomerase genes copy number in newly diagnosed childhood acute lymphoblastic leukemia. AB - Topoisomerase genes were analyzed at both DNA and RNA levels in 25 cases of newly diagnosed childhood acute lymphoblastic leukemia (ALL). The results of molecular analysis were compared to risk group classification of children in order to identify molecular characteristics associated with response to therapy. At diagnosis, allelic imbalance at topo-isomerase IIalpha (TOP2A) gene locus was found in 75% of informative cases whereas topoisomerase I and IIbeta gene loci are altered in none or only one case, respectively. By semi-quantitative Polymerase chain reaction, we found a 2.5 to 8-fold TOP2A gene amplification in 72% of the children, which was correlated to gene overexpression in every case. These results show that TOP2A gene amplification is a frequent event in ALL at diagnosis. Interestingly, we also identified a small population of children that do not present TOP2A gene amplification or gene overexpression and who are significantly associated with very high risk classified patients showing glucocorticoid resistance. In conclusion, characterization of TOP2A gene status in childhood ALL at diagnosis provides useful complementary information for risk assessment. PMID- 12646942 TI - Urolithiasis in pediatric patients with acute lymphoblastic leukemia. AB - We evaluated the incidence, timing, and consequences of urolithiasis in children with acute lymphoblastic leukemia (ALL). A total of 20 patients with urolithiasis were identified from 2095 patients with ALL treated at St Jude Children's Research Hospital on consecutive protocols between 1968 and 1998. For remission induction therapy, all patients received daily prednisone; continuation chemotherapy regimens differed by protocol with some including pulses of prednisone or dexamethasone and others no glucocorticoid. Patients with urolithiasis were older at diagnosis of ALL than those without urolithiasis (median age, 7.5 vs 5.0 years; P=0.03) and less likely to be black (P=0.03) than white or Hispanic, but sex and treatment era did not differ. Presenting symptoms included abdominal or flank pain, hematuria, and dysuria. All stones analyzed biochemically were calcium stones. The incidence of urolithiasis after completion of therapy was 1.8 per 10 000 person-years. Compared to this baseline rate, the relative risk of urolithiasis was 45 (P<0.01) during induction therapy, 22 (P<0.01) during continuation therapy with glucocorticoids, and 5.1 (P>0.05) during continuation therapy without glucocorticoids. Urolithiasis occurred 4.5 times more often during continuation treatment with glucocorticoids than without (P<0.05). Seven patients (35%) had recurrent urolithiasis. Patients with ALL are at risk of developing calcium renal stones during chemotherapy, especially when a glucocorticoid is included. PMID- 12646943 TI - Amplification of AML1 on a duplicated chromosome 21 in acute lymphoblastic leukemia: a study of 20 cases. AB - This study identifies multiple copies of the AML1 gene on a duplicated chromosome 21, dup(21), as a recurrent abnormality in acute lymphoblastic leukemia (ALL). Clusters of AML1 signals were visible at interphase by fluorescence in situ hybridization (FISH). In metaphase, they appeared tandemly duplicated on marker chromosomes of five distinct morphological types: large or small acrocentrics, metacentrics, submetacentrics or rings. The markers comprised only chromosome 21 material. Karyotypes were near-diploid and, besides dup(21), no other established chromosomal changes were observed. A total of 20 patients, 1.5 and <0.5% among consecutive series of childhood and adult ALL respectively, showed this phenomenon. Their median age was 9 years, white cell counts were low and all had a pre-B/common immunophenotype. Although this series is not the first report of this abnormality, it is the largest, permitting a detailed description of the variety of morphological forms that duplicated chromosome 21 can assume. PMID- 12646945 TI - Telomerase inhibition enhances apoptosis in human acute leukemia cells: possibility of antitelomerase therapy. AB - Telomerase is a ribonucleoprotein enzyme that maintains protective structures at the ends of eukaryotic chromosomes. We examined the impact of telomerase inhibition by the dominant-negative human catalytic subunit of telomerase (DN hTERT) on the biological features of acute leukemia. We introduced vectors encoding dominant- negative (DN)-hTERT, wild-type (WT)-hTERT, or a control vector expressing only a drug-resistant marker into a telomerase-positive human acute lymphoblastic leukemia cell line, HAL-01. Expression of DN-hTERT dramatically inhibited telomerase activity, leading to apoptotic cell death. Mutant telomerase expression also enhanced daunorubicin-induced apoptosis. Nude mice (n=5 per group) received subcutanous implants of HAL-01 cells expressing the control vector or DN-hTERT or WT-hTERT. Implantation of HAL-01 cells expressing control vector (n=5) rapidly produced tumors, whereas implantation of those expressing DN hTERT (n=5) did not. Thus, telomerase inhibition both growth of HAL-01 cells in vitro and tumorigenic capacity in vivo. Furthermore, the G-quadruplex-interactive telomerase-specific inhibitor, telomestatin, shortened the telomere length and induced apoptosis in freshly isolated primary acute leukemia cells. These results suggest that antitelomerase therapy may be useful in some acute leukemias in combination with antileukemic agents such as daunorubicin. PMID- 12646944 TI - A phase II study of alpha-interferon and oral arabinosyl cytosine (YNK01) in chronic myeloid leukemia. AB - YNK01 (Starasid) is a prodrug that is adsorbed in the gut and is transformed in the liver in arabinosyl cytosine (AC). Low-dose AC (LDAC) is useful for the treatment of Philadelphia positive (Ph+) chronic myeloid leukemia (CML), especially in combination with alpha-interferon (alphaIFN). The use of YNK01 can avoid the daily s.c. injection of conventional AC. To assess the safety and the efficacy of alphaIFN and YNK01, we enrolled 86 consecutive previously untreated chronic phase Ph+ CML patients in a phase II study of alphaIFN (Intron-A) 5 MIU/m(2) daily and YNK01 600 mg daily 14 days a month. The 6-month complete hematologic response and the 12-month major cytogenetic response rates were 78 and 28%, respectively. In a prior study of alphaIFN and conventional LDAC, they were 62 and 22%, respectively. However, the compliance to the treatment was poor, with 25% of cases discontinuing the treatment within the first year. This was not because of the severity of the side effects but because of the frequency, duration and repetition of the side effects, for an overall frequency of 13.17 adverse events, mostly grade 1 and 2, per patient per year. Therefore, the study of this effective combination is being pursued, testing lower doses of alphaIFN and YNK01. PMID- 12646946 TI - Fibrates and medroxyprogesterone acetate induce apoptosis of primary Burkitt's lymphoma cells and cell lines: potential for applying old drugs to a new disease. AB - Current therapies for Burkitt's lymphoma (BL) utilise combined cytotoxic chemotherapy, but these treatments are not always available in areas where the disease is endemic and are also markedly less successful in AIDS-related BL. Therefore, additional therapies are urgently required. We demonstrate here that combined fibrates and MPA exert powerful, antiproliferative actions against well characterised Daudi, Raji and L3055 BL cell lines and primary BL cells. Detailed studies in L3055 demonstrated that this activity was mediated by induced apoptosis and confirmed by observations that overexpression of the antiapoptotic genes bcl-2 or bcl-x(L) conferred significant protection against the drugs. Importantly, since fibrates and MPA are inexpensive and stable with minimal associated toxicities, we suggest that these drugs should be considered as adjuncts to currently available treatments for BL in endemic and AIDS-related disease. PMID- 12646947 TI - High susceptibility of human leukemic cells to Fas-induced apoptosis is restricted to G1 phase of the cell cycle and can be increased by interferon treatment. AB - In this study, we analyzed the influence of cell cycle status manipulations of leukemic cells on Fas-mediated apoptosis using the GM-CSF-dependent human myeloid leukemia cell line AML-193 as a model. GM-CSF and long-term treatment with interferon-gamma (IFN-gamma) or interferon-alpha (IFN-alpha) were used to manipulate the cell cycle status. Control cells were GM-CSF deprived, nonproliferating cells. IFN-gamma or IFN-alpha treatment did not induce proliferation in control cells, but resulted in recruitment of cells from resting G(0) phase into activated G(1) phase. Using agonistic anti-Fas antibodies (FAS18), we demonstrated that this shift from G(0) to G(1) was accompanied by a 2.5-fold increase in Fas sensitivity. A similar increase in sensitivity to FAS18 could be obtained by induction of proliferation with GM-CSF. Quantitative FACS analysis of surviving cells after FAS18-induced apoptosis showed deletion of the G(1) compartment, but complete protection of resting G(0) cells. Cells in S or G(2)/M phase were relatively protected against Fas induction. In conclusion, sensitivity to Fas-mediated apoptosis was restricted to cells in G(1) phase of the cell cycle, and can be increased by treatment of cells with interferons. By this mechanism, interferon treatment may render leukemic cells more susceptible to lysis by T cells during immunotherapeutic interventions. PMID- 12646948 TI - Prospective analysis of minimal bone marrow infiltration in pediatric Burkitt's lymphomas by long-distance polymerase chain reaction for t(8;14)(q24;q32). AB - The chromosomal translocation t(8;14)(q24;q32) represents a characteristic marker for Burkitt's lymphoma (BL). This translocation involves the MYC oncogene on chromosome 8 and the immunoglobulin heavy-chain (IgH) locus on chromosome 14. Since the translocation does not produce a fusion gene, we established a long distance polymerase chain reaction (LD-PCR) assay that can detect the t(8;14) at the genomic level. The sensitivity of the LD-PCR was 10(-4). We used the LD-PCR assay to prospectively study 78 BL patients and found a specific PCR product in 52 of them. Among the 52 positive patients, we could test both the tumor and the bone marrow (BM) at diagnosis in 33 and determined the prevalence of minimal disseminated disease (MDD) at diagnosis. In 12/33 patients, BM was positive by LD PCR and in 10 of them we conducted a study of minimal residual disease (MRD). Eight out of 10 children showed a clearance of MRD after one cycle of chemotherapy. The only two patients who did not achieve a negative MRD status died of disease progression. The comparative analysis of sensitivity of BM aspirate, BM biopsy and LD-PCR in t(8;14)-positive patients demonstrated a superiority of the molecular method in the assessment of MDD. The LD-PCR for t(8;14) is an important tool to study minimal BM infiltration at diagnosis and to determine its response kinetics in BL. PMID- 12646949 TI - Involvement of PI3K/Akt pathway in cell cycle progression, apoptosis, and neoplastic transformation: a target for cancer chemotherapy. AB - The PI3K/Akt signal transduction cascade has been investigated extensively for its roles in oncogenic transformation. Initial studies implicated both PI3K and Akt in prevention of apoptosis. However, more recent evidence has also associated this pathway with regulation of cell cycle progression. Uncovering the signaling network spanning from extracellular environment to the nucleus should illuminate biochemical events contributing to malignant transformation. Here, we discuss PI3K/Akt-mediated signal transduction including its mechanisms of activation, signal transducing molecules, and effects on gene expression that contribute to tumorigenesis. Effects of PI3K/Akt signaling on important proteins controlling cellular proliferation are emphasized. These targets include cyclins, cyclin dependent kinases, and cyclin-dependent kinase inhibitors. Furthermore, strategies used to inhibit the PI3K/Akt pathway are presented. The potential for cancer treatment with agents inhibiting this pathway is also addressed. PMID- 12646950 TI - Inhibition of human leukemia in an animal model with human antibodies directed against vascular endothelial growth factor receptor 2. Correlation between antibody affinity and biological activity. AB - Vascular endothelial growth factor (VEGF) and its receptors (VEGFR) have been implicated in promoting solid tumor growth and metastasis via stimulating tumor associated angiogenesis. We recently showed that certain 'liquid' tumors such as leukemia not only produce VEGF, but also express functional VEGFR, resulting in an autocrine loop for tumor growth and propagation. A chimeric anti-VEGFR2 (or kinase insert domain-containing receptor, KDR) antibody, IMC-1C11, was shown to be able to inhibit VEGF-induced proliferation of human leukemia cells in vitro, and to prolong survival of nonobese diabetic-severe combined immune deficient (NOD-SCID) mice inoculated with human leukemia cells. Here we produced two fully human anti-KDR antibodies (IgG1), IMC-2C6 and IMC-1121, from Fab fragments originally isolated from a large antibody phage display library. These antibodies bind specifically to KDR with high affinities: 50 and 200 pM for IMC-1121 and IMC 2C6, respectively, as compared to 270 pM for IMC-1C11. Like IMC-1C11, both human antibodies block VEGF/KDR interaction with an IC(50) of approximately 1 nM, but IMC-1121 is a more potent inhibitor to VEGF-stimulated proliferation of human endothelial cells. These anti-KDR antibodies strongly inhibited VEGF-induced migration of human leukemia cells in vitro, and when administered in vivo, significantly prolonged survival of NOD-SCID mice inoculated with human leukemia cells. It is noteworthy that the mice treated with antibody of the highest affinity, IMC-1121, survived the longest period of time, followed by mice treated with IMC-2C6 and IMC-1C11. Taken together, our data suggest that anti-KDR antibodies may have broad applications in the treatment of both solid tumors and leukemia. It further underscores the efforts to identify antibodies of high affinity for enhanced antiangiogenic and antitumor activities. PMID- 12646951 TI - Chimerism testing after allogeneic stem cell transplantation: importance of timing and optimal technique for testing in different clinical-biological situations. PMID- 12646952 TI - Serial quantification of lymphoid and myeloid mixed chimerism using multiplex PCR amplification of short tandem repeat-markers predicts graft rejection and relapse, respectively, after allogeneic transplantation of CD34+ selected cells from peripheral blood. AB - We used multiplex amplification of nine microsatellite sequences (PCR-STR) to analyse chimerism in pure populations of T cells and neutrophils from peripheral blood from 40 patients submitted to an allogeneic transplant, 22 having received a T-cell depleted (TCD) peripheral blood graft by means of CD34(+) selection (allo-PBT/CD34(+)), and 18, an unmodified graft (allo-SCT; 13 allogeneic bone marrow transplants and five allo-PBT). T-cell mixed chimerism (TcMC) was observed in 16 of the 22 (72.3%) patients receiving an allo-PBT/CD34(+), but in only one of the 18 (5.5%) patients receiving an allo-SCT (P=0.0001). TcMC was transient (n=6), stable (n=7), and associated with poor haematopoietic engraftment (n=4). All patients with TcMC who developed graft failure had more than 30% of host T cells. Myeloid MC (MyMC) was observed in four (19%) allo-PBT/CD34(+) patients and in three (17%) allo-SCT patients (P=NS). Five out of seven (71%) patients with MyMC relapsed, all of them diagnosed with myeloid malignancies, as compared with two of the 20 (10%) patients with complete donor chimerism (P&<0.0001). In conclusion, TcMC appears in a significant number of allo-PBT/CD34(+) patients and may be associated with poor engraftment when the percentage of host T cells is >30%; likewise, MyMC appears in a small percentage of recipients of both allo PBT/CD34(+) and allo-SCT patients, and is associated with leukaemia relapse in myeloid malignancies. PMID- 12646953 TI - Quantification of donor and recipient hemopoietic cells by real-time PCR of single nucleotide polymorphisms. AB - Analysis of changes in recipient and donor hemopoietic cell origin is extremely useful to monitor the effect of stem cell transplantation (SCT) and sequential adoptive immunotherapy by donor lymphocyte infusions (DLI). We developed a sensitive and accurate method to quantify the percentage of recipient and donor cells by real-time PCR using single nucleotide polymorphisms (SNPs) as markers. Allele-specific PCR of seven SNPs resulted in specific markers for donor or recipient in 97% of HLA-identical sibling pairs. Both, recipient- and donor derived hemopoietic cells can be simultaneously analyzed in 67% sibling pairs. We expect this can be increased to approximately 99% by developing three additional SNP-PCR. Serial dilution of SNP-positive DNA into either SNP-negative DNA or water revealed a detection limit of 0.1-0.01% depending on the amount of input DNA and start C(t) of the used SNP-PCR. Application of our real-time SNP-PCR method for a CML patient treated by allogeneic SCT and DLI demonstrated its feasibility to follow donor T-cell chimerism and early detection of residual and recurrent autologous hemopoiesis in response to treatment. This detailed monitoring of the genetic origin of hemopoietic cells, in particular immune effector cells and target cells after SCT and DLI, may substantially contribute to understanding of the mechanisms that play a role in the success of treatment. PMID- 12646954 TI - Quantification of donor and recipient hemopoietic cells by real-time PCR of single nucleotide polymorphisms. PMID- 12646955 TI - Fatal myelodysplastic syndrome developing during therapy with imatinib mesylate and characterised by the emergence of complex Philadelphia negative clones. PMID- 12646956 TI - Prognostic impact of t(9;11) in childhood acute myeloid leukemia (AML). PMID- 12646957 TI - TET1, a member of a novel protein family, is fused to MLL in acute myeloid leukemia containing the t(10;11)(q22;q23). PMID- 12646958 TI - VLA-4 affinity correlates with peripheral blood white cell count and DNA content in patients with precursor B-ALL. PMID- 12646960 TI - Expression of retinoic acid receptor-target genes during retinoic acid therapy for acute promyelocytic leukemia. PMID- 12646959 TI - A discriminating screening is necessary to ascertain EVI1 expression by RT-PCR in malignant cells from the myeloid lineage without 3q26 rearrangement. PMID- 12646961 TI - P-glycoprotein (P-gp) and multidrug resistance-associated protein 1 (MRP1) are induced by arsenic trioxide (As(2)O(3)), but are not the main mechanism of As(2)O(3)-resistance in acute promyelocytic leukemia cells. PMID- 12646964 TI - Expression of the two alternative [a] and [b] transcripts of CCND1 gene in cyclin D1-expressing B-malignancies: relevance for the pathogenesis. PMID- 12646965 TI - Hairy cell leukemia and Lambert-Eaton myasthenic syndrome. PMID- 12646962 TI - Molecular monitoring of acute myeloid leukemia associated with inv(16): threshold of CBFbeta/MYH11 transcript copy number above which relapse occurs and below which continuous Complete Remission is likely. PMID- 12646968 TI - [Myelodysplastic syndromes (preleukemic states)]. PMID- 12646969 TI - [NSAIDs caused gastric mucosal injury: with a special reference to COX-2]. AB - Nonsteroidal antiinflammatory drugs (NSAIDs) have been shown to cause mucosal injury in the gastrointestinal tract as a side effect, occasionally turning out to be severe complications such as bleeding and perforation. So far NSAIDs-caused mucosal injury was attributed to their inhibitory effects on the activity of cyclooxygenase-1 (COX-1) which is expressed and shown to play a crucial role for the mucosal protection via producing prostaglandin E2 in the stomach. However, a recent progress of the understanding about COX physiology has revealed that NSAIDs cause gastric mucosal injury by inhibiting not only COX-1 but also COX-2 in the stomach. COX-1 inhibition alone has been demonstrated not to cause gastric mucosal injury. In addition, a selective COX-2 inhibitor which is demonstrated to have much less harmful effect in the stomach is now widely used as a safer NSAID in USA. Moreover, a selective COX-2 inhibitor is recently considered to have an inhibitory effect on growth of a certain type of cancers, thereby being in the spotlight as a chemopreventive agent. PMID- 12646970 TI - Effect of the surface potential of the hemodialysis membrane and the electrical charge of the gadolinium contrast medium on dialyzability. AB - PURPOSE: To search for methods of improving the excretion of injected gadolinium contrast medium (GdCM) in hemodialysis (HD) patients, we investigated the effect of the surface potential of HD membranes and the electrical charge of GdCM on the dialyzability of GdCM. MATERIALS AND METHODS: Ionic (Gd-DTPA) or non-ionic (Gd DO3A) GdCM solutions were dialyzed using a clinical HD unit. Two types of HD membranes, AN69 with a surface potential and PMMA without, were used. GdCM clearance was then calculated. RESULTS: Gd-DTPA clearance was significantly higher for PMMA membranes than for AN69 membranes. Gd-DO3A clearance was slightly higher for AN69 membranes than for PMMA membranes. The difference in Gd-DTPA and Gd-DO3A clearance values was not significant when PMMA membranes were used. CONCLUSION: These data indicate that non-ionic GdCM is preferable to ionic GdCM in patients receiving dialysis through an electrically positive membrane. Either ionic or non-ionic GdCM can be used when a normal dialysis membrane is being used. PMID- 12646971 TI - Preoperative administration of methylprednisolone attenuates cytokine-induced respiratory failure after esophageal resection. AB - Proinflammatory cytokines have been implicated in mediating respiratory failure associated with major surgery. We investigated the effect of giving glucocorticoids preoperatively for the prophylaxis of surgical stress and the association of cytokine levels, such as interleukin-6 (IL-6) and interleukin-8 (IL-8), with oxygenation after esophagectomy. We studied 17 patients who underwent subtotal esophagectomy. Seven patients (steroid group) were chosen at random to be given methylprednisolone (10 mg/kg) and 10 patients (control group) to be given saline intravenously before operation. Plasma and bronchoalveolar lavage fluid (BALF) IL-8 levels in the control group were significantly higher than those in the steroid group. In both groups, plasma IL-6 levels were significantly higher than those in BALF, but in contrast, BALF IL-8 levels were significantly higher than plasma levels of IL-8 postoperatively. The PaO(2)/FiO(2) ratio was significantly reduced in the control group. The PaO(2)/FiO(2) ratio of the control group had significantly lower values than that of the steroid group. There was significant correlation between BALF IL-8 levels and the PaO(2)/FiO(2) ratio postoperatively. We conclude that preoperative administration of methylprednisolone may attenuate postoperative reduction of arterial oxygen saturation by suppressing the release of cytokines. PMID- 12646972 TI - Clinicopathological characteristics of 10 patients with rupture of both ventricular free wall and septum (double rupture) after acute myocardial infarction. AB - Cardiac ruptures after myocardial infarction are classified as ventricular free wall ruptures (FWR), ventricular septal ruptures (VSR), and papillary muscle ruptures (PMR). A combination of any two types of rupture is called "ventricular double rupture;" (VDR) and shows a specific clinical course. 3,284 patients with acute myocardial infarction (AMI) were admitted to the CCU of our hospital between April, 1973 and December, 2001, and 10 patients (8 males and 2 female, aged 54 through 82 years) with VDR were clinicopathologically evaluated. All were diagnosed as VDR consisting of FWR and VSR. VDR was observed in 0.30% of all patients with AMI, in 3.0% of those with FWR, and in 16.1% of those with VSR. The infarct site was anteroseptal in 3 patients, anterolateral in 3, inferior in 3, and posterolateral in 1. Two patients with inferior infarction complicated RV infarction and a patient with posterolateral infarction had healed inferior infarction. The risk factors related to VDR were age, a history of hypertension, increased sympathetic tone to improve hemodynamic aggravation after perforation, cardiotonic agents, thrombolytic agents, delayed reperfusion, right ventricular volume overload by shunt and re-infarction. However, these factors might have played only a subsidiary role. The most important factor in VDR was the pathological findings. The site of septal perforation was the apex close to the septum-free wall junction in 9 patients and the site of rupture was also apical in 8 patients. Four patients already had VSR on admission to our CCU. FWR developed soon after VSR was demonstrated in 4 patients. FWR and VSR occurred simultaneously in one patient. These results suggest that VSR in the apical region is a precursor of VDR and requires the earliest surgical treatment. Surgical treatment was carried out in the operating room in 5 patients and 3 (60.0%) of them survived for 4 months or more. Two patients with rupture incidentally detected during operation for VSR were discharged and are still alive, though another one with free wall blow out rupture died 129 days after operation. Bedside thracotomy was performed in 3 patients and all of them died. PMID- 12646973 TI - Prognostic suggestion in the evaluation of solid component in poorly differentiated adenocarcinoma of the lung. AB - Pulmonary adenocarcinoma composed of pure or predominant solid components is reported to be a highly malignant tumor. However, the existence of solid components and its connection with biological behavior have not been well documented. To answer this question, we histologically subclassified poorly differentiated adenocarcinoma (P/D Ad Ca) into solid type and non-solid type, and compared the biological behavioral characteristics. MATERIAL AND METHODS: All histological specimens of surgically resected primary lung carcinoma diagnosed as P/D Ca or large cell carcinoma in Nippon Medical School Hospital were re evaluated according to the 1999 WHO manual. The cases re-evaluated as P/D Ad Ca were further divided into solid type and non-solid type according to our original definition: the solid type contains solid components where a glandular structure is not recognized in more than one high-power field, while in the non-solid type, a small glandular structure is observed in every high-power field. The differences in the occurrence of lymph node metastasis were assessed by Fisher's exact test. RESULTS: Among 109 cases satisfying both histological and clinical investigation, 45 cases were re-evaluated as P/D Ad Ca; solid type (n=22), and non-solid type (n=23). Lymph node metastases occurred at a higher rate in the solid type than in the non-solid type (p<0.01). CONCLUSION: Patients with solid type Ad Ca have reached a more advanced stage than patients having non-solid type due to high metastatic rate to lymph nodes. These results suggest that we should not overlook solid components even if the solid components are the focal lesion. This sub-typing alerts clinicians to survey metastases, and may contribute to therapeutic strategies in the future. PMID- 12646974 TI - Surgical therapy in Hashimoto's thyroiditis. AB - Hashimoto's thyroiditis (HT) is usually considered to be manageable by levothyroxine (L-T4) administration, which can reduce the thyroid volume and supplement the lack of hormone. However, we sometimes encounter a huge goiter that has not shrunk in response to L-T4 therapy. These goiters continue to produce symptoms of compression and an unsightly appearance. Here we discuss the surgical indication and procedure for HT. Thirteen patients with clinically diagnosed Hashimoto's thyroiditis involving a huge diffuse goiter that produced pressure symptoms or nodular lesions were treated with surgery. The gender, age distribution, total dose and period of L-T4 administration prior to the operation, and clinical symptoms caused by the large goiter were evaluated in each case. The titer of antibodies was extremely elevated in 8 HT patients with a diffusely enlarged goiter. The total period of L-T4 medication ranged from 6 to 25 years. A subtotal thyroidectomy in which a small amount of thyroid was left in the posterior area of the bilateral lobes was performed in the 8 cases of diffusely enlarged goiter. Pressure symptoms and the unsightly appearance caused by the goiter were relieved by the surgery in all cases. No surgical complications developed. In conclusion, the surgery is an effective therapy for HT patients who have persistent compression symptoms and/or an unsightly neck appearance due to a large goiter despite long-term L-T4 treatment. PMID- 12646975 TI - Association of ovarian tumor epithelium coexpressing HLA-DR and CA-125 antigens with tumor infiltrating cytotoxic T lymphocytes. AB - Understanding of the relationship between tumor infiltrating cytotoxic T lymphocytes (CD 8(+)TILs) and tumor cells as well as tumor-associated antigens is important and may reflect the extent of immune response of the patient to the tumor, thus providing a useful clue relevant to the prognosis. The purpose of this study was to determine the relationship between the expression of HLA-DR and CA-125 antigens and the presence of CD 8(+)TILs with regard to the established clinicopathological factors of ovarian carcinomas using immunohistochemical methods. Thirty-one ovarian carcinomas consisting of 20 serous, 7 mucinous, and 4 clear cell types were examined. Of these, 18 (58%) and 22 (71%) were positive for HLA-DR and CA-125 antigens respectively, and the overall mean number of the CD 8(+)TILs was 7.2+/-2.9. A significant association was observed between the mean number of CD8(+)TILs and tumor grade (P=0.01), disease stage (P=0.003), and patient outcome (P=0.01). The mean number of CD 8(+)TILs in HLA-DR positive (8.6+/-2.2) or CA-125 positive (8.4+/-2.1) tumors was significantly higher than that in HLA-DR negative (5.2+/-2.5; P=0.0003) or CA-125 negative (4.2+/-2.2; P=0.00002) tumors. These significant levels were further enhanced by one order of magnitude when the mean number of CD 8(+)TILs in tumors positive for both HLA-DR and CA-125 antigens (9.1+/-1.7) was compared to that in HLA-DR negative or CA-125 negative tumors. The frequency of cancer-related deaths in HLA-DR and CA-125 positive tumors was significantly lower than in the negative tumors (P=0.01). These data suggest that concurrent expression of HLA-DR and CA-125 antigens may augment the immune response of the patient to the tumor, thus providing a potential clue relevant to the prognosis. PMID- 12646976 TI - Genetic diagnosis of Werdnig-Hoffmann disease: a problem for application to prenatal diagnosis. AB - We report a floppy infant with Werdnig-Hoffmann disease (spinal muscular atrophy: SMA type 1) and Klinefelter syndrome. After genetic counseling with parents, a genetic diagnosis using DNA from the infant's peripheral blood mononuclear cells was performed. The parents' deletion of exons 7 and 8 of the survival motor neuron (smn) gene and exons 4 and 5 of the neuronal apoptosis inhibitory protein (naip) gene were noted in the infant, so he was confirmed to have SMA type 1. The parents wanted to receive a prenatal diagnosis on the next pregnancy. However this genetic test is achieved by confirming that a specific band can not be detected by PCR. Therefore, this method should be applied with great care to prenatal diagnosis using chorionic villi, which may be contaminated with maternal tissue. PMID- 12646977 TI - Behcet's syndrome associated with acute myocardial infarction. AB - We report here a case of Behcet's syndrome. The patient was suddenly affected by chest pain during the clinical course of the disease and developed abdominal pain and melena after 5 weeks. The diagnosis of acute anteroseptal myocardial infarction was made on the basis of the electrocardiography findings, and many perforated ulcers were confirmed in specimens of the resected colon. Coronary angiography showed constriction by 50% of the left anterior descending artery. There were no atherosclerotic changes. Pulse therapy was performed using sodium methylprednisolone succinate at a dose of 1,000 mg for 3 days, followed by oral administration of prednisolone at a dose of 60 mg/day, leading to improvement of all the symptoms after 3 weeks. The electrocardiogram findings at that time were normal. Prompt resolution of the symptoms provided by corticosteroid therapy supports the conclusion of previous studies that myocardial infarction in Behcet's syndrome may possibly be due to vasculitis of the coronary arteries. In Behcet's syndrome, myocardial infarction is rare but should be considered as one of the most important lesions that determine the prognosis. PMID- 12646978 TI - [Pharmacoeconomical evaluation of clinical pathway in gastrectomy patients]. AB - The aim of the present study was to ascertain the pharmacoeconomical efficacy of a clinical pathway (CP) employing medication management and instruction tasks (i. e. pharmaceutical care and counseling for inpatients) in gastrectomy patients. Pharmaceutical services of a uniform quality were provided. These included a CP check sheet, medication management, and a history of the drugs chiefly prescribed by pharmacists. As a result, the average number of hospitalized days among the patients who were offered pharmaceutical care compared with those who were not was significantly shortened from 35.4 days to 26.1 days (P<0.001). Moreover, the average cost of medication was also significantly reduced from 270,631 yen to 190,331 yen (P<0.05). These data provide the first evidence that a CP employing medication management and instruction tasks for gastrectomy patients may play a substantial role in saving on medical costs. PMID- 12646979 TI - [Endocrine surgery. The ninth report: the latest data on and clinical characteristics of the epidemiology of thyroid carcinoma]. AB - Thyroid disease is comparatively prevalent, and it is important to perform careful palpation of the anterior neck when patients are first examined. In this paper, the latest data on and clinical characteristics of the epidemiology of thyroid carcinoma are described. According to Japanese cancer statistics, in 1996, the number of cases of thyroid carcinoma was 1,390 men and 5,437 women. The female to male ratio was 3.91: 1. The crude rate of thyroid carcinoma per 100,000 population was 2.3 men and 8.5 women. The age-adjusted incidence rate showed a gradual increase with age, and the peak of the age-adjusted incidence rate was over 80 years old among men, and from 70 to 80 years old among women. According to the Japanese Thyroid Cancer Registration Committee, from 1977 to 1999, papillary thyroid carcinoma accounted for more than 85% (30,256) of thyroid carcinomas among 35,312 patients. Medullary thyroid carcinoma (1.4%) and anaplastic thyroid carcinoma (1.6%) were rare. The female to male ratio of thyroid carcinoma was 6.1: 1. Registration was made in accordance with the general rules issued by the Japanese Society of Thyroid Surgery. According to the vital statistics, mortality related to thyroid carcinoma among all cancer-related deaths was 0.23% among men (411) and 0.76% among women (887) in 2000. The female to male ratio of mortality was 2.16: 1. The peak of mortality was from 70 to 80 years old among men, and from 80 to 90 years old among women. Most patients with differentiated thyroid carcinoma had a low mortality rate, but anaplastic carcinoma had a poor prognosis. Lung and bone were the main distant metastatic sites, and respiratory insufficiency was the most common specific fatal condition. PMID- 12646980 TI - [Stem cell transplantation in the 21st century]. PMID- 12646981 TI - [Nerve block therapy for intractable pain: 3 cases of erythlomelalgia]. AB - Erythromelalgia is a rare condition that is characterized by the triad of redness, increased temperature, and intense pain of feet or hands. This syndrome was first described in 1878, however, universal classification, precise mechanism and successful medical treatment of erythoromelalgia have not been described. We experienced 3 cases of erythomelalgia in our outpatient pain clinic that showed the different progress. One patient experienced the long-term pain complicated with neuropathic pain. The pain of other two patients was eliminated early by nerve block treatments, and they did not showed chronic pain. From the prophylactic viewpoint of chronic pain, the most considerable matter is early and certain elimination of severe pain under certain diagnosis. In this article, the mechanisms of passing into the chronic pain state as well as the efficacy of neural blockade for intractable pain such as erythromelalgia are discussed. PMID- 12646982 TI - Ultrastructure of normal rat hepatocytes: stereological arrangement of the organellae. PMID- 12646983 TI - From pain research to pain treatment: role of human pain models. AB - There is no objective measure of pain; we can however measure different aspects of the pain perception. Earlier experimental pain models often only involved induction of cutaneous pain. Recently new experimental models have been developed eliciting deep muscle and visceral pain that may more closely resemble the clinical pain condition. It is imperative to use multi-modal and multi-structure pain induction and assessment techniques, as a simple model cannot describe the very complex and multi-factorial aspects of clinical pain. The importance of peripheral and central hyperexcitability for acute and chronic pain has been demonstrated in animals and to some extent in humans. But in spite of our immense knowledge we still do not know how to prevent and treat this hyperexcitability. Our understanding of nociceptive mechanisms involved in acute and chronic pain and the effects of anaesthetic drugs or combinations of drugs on these mechanisms in humans may also be expanded with experimental human models. This knowledge can then help us to develop and test therapeutic regimes in patients with acute and chronic pain. PMID- 12646984 TI - Clinical significance of wide QRS complexes at the termination of paroxysmal supraventricular tachycardias. AB - BACKGROUND: A wide QRS complex is not a rare electrocardiographic phenomenon at the termination of paroxysmal supraventricular tachycardia (PSVT), but no plausible underlying mechanism has yet been proposed. The purpose of the present study was to elucidate the frequency and the underlying mechanism of the wide QRS complexes at the termination of PSVT. METHODS: We retrospectively reviewed 305 electrocardiograms (ECGs) from 100 patients, on which PSVT termination was recorded. The frequency of the wide QRS complexes was analyzed in 181 ECGs to avoid duplication, because there were 124 ECGs obtained from the same patients with same methods. The 181 ECGs were divided by morphology into three groups: Type A, termination with wide QRS complex without pause; Type B, wide QRS complex following initial pause after termination; Type C, wide QRS complex following the first narrow QRS after termination. RESULTS: The wide QRS complex was recorded in 81/181 (44.8%) ECGs (Type A; 3/81 (3.7%), Type B; 44/81 (54.3%), Type C; 62/81 (55.6%) ) and its frequency was not dependent on the mechanism of PSVT. It was more frequently observed after a long pause, and was frequently induced by procedures that increase vagal tone, such as intravenous adenosine 5' triphosphate administration (16/22: 72.7%) and vagal stimulation maneuvers (16/32: 50%). There were a total of 41 wide QRS complexes (44.6%) which had a preceding sinus P wave, out of a total of 92 wide QRS complexes in all three types. These 41 wide QRS complexes included 30/44 (68.2%) Type B wide QRS, and 11 (24.4%) Type C wide QRS complexes. CONCLUSION: The aberrant conduction or escaped ventricular contraction was suggested to be the underlying mechanism of the majority of wide QRS complexes and ventricular premature contraction is less frequent. PMID- 12646985 TI - Windows of therapeutic opportunity on fetal growth retardation induced by transient intrauterine ischemia in rats. AB - OBJECTIVE: To assess the windows of therapeutic opportunity for drugs with various chemical actions on fetal growth retardation induced by transient intrauterine ischemia in rats. METHODS: At 17 days of gestation, ischemia was induced by 30 min of right uterine artery occlusion. The administration of either alpha-phenyl-N-tert-butyl-nitrone (PBN), FK 506, nifedipine, or MK-801 to pregnant rats was randomly started before occlusion, 1 hour, 3 hours, or 24 hours after recirculation. All of the pups were delivered by cesarean section at 21 days of gestation and were weighed to determine the degree of fetal growth retardation. RESULTS: The vehicle-treated animals exposed to ischemia showed a significant decrease in fetal body weight compared with the normoxic control animals. The growth disturbances were prevented by nifedipine and MK-801 only when given just prior to ischemia. In contrast, PBN and FK 506 had a protective effect even when given 1 hour and 3 hours after the start of recirculation, respectively. CONCLUSIONS: The present results indicate that treatment with PBN and FK 506 gives relatively wide windows of therapeutic opportunity in fetal growth retardation induced by transient intrauterine ischemia in rats and suggest the possibility of therapeutic intervention after the start of recirculation. PMID- 12646986 TI - Effect of regular maternal exercise on lipid peroxidation levels and antioxidant enzymatic activities before and after delivery. AB - This study was designed to investigate the influence of maternal aerobic exercise on lipid peroxidation levels and antioxidant enzymatic activities before and after delivery. Predelivery and 1- and 24-hour post partum blood samples were collected from 18 nulliparous healthy pregnant women who exercised regularly throughout the whole period of pregnancy and from 22 matched controls. The plasma concentration of malondialdehyde (MDA) was measured as an indicator of lipid peroxidation. Erythrocyte enzymes, superoxide dismutase (SOD), glutathione peroxidase (GSHPx) and catalase (CAT), were measured as intracellular antioxidant markers. In the control group, MDA increased slightly from predelivery to 1 hour post partum and then increased significantly at 24 hours post partum, with significant increase in SOD and CAT activities. On the other hand, MDA of the exercise group remained unaltered at 1 and 24 hours after delivery. At 1 hour post partum, the SOD and CAT levels of the exercise group increased markedly and then decreased to predelivery levels. The present results indicate that uncontrolled lipid peroxidation occurs during labor and suggest that continuing regular maternal exercise may reduce labor-induced lipid peroxidation by improving the defense capabilities against free radical generation. PMID- 12646987 TI - Relationship between grade of fibrosis in pancreatic stump and postoperative pancreatic exocrine activity after pancreaticoduodenectomy: with special reference to insufficiency of pancreaticointestinal anastomosis. AB - BACKGROUND: Pancreaticoduodenectomy (PD) has become a standard operation for malignant and benign periampullary diseases. Although the operative mortality of PD has decreased to less than 4% in hospitals with many cases of PD, the leakage of pancreaticointestinal anastomosis (PIA) still carries a substantial risk of lethal outcome. The aim of this study was to evaluate the local factors that affect the incidence of PIA leakage by evaluation of exocrine function and fibrosis in the pancreatic remnant following PD. METHOD: Twenty-eight patients (17 pancreatic disease, 8 bile duct cancers and 3 ampullary cancers) underwent PD with complete extracorporeal pancreatic juice drainage. The cut-end of the pancreatic remnant was histologically studied for its grade of fibrosis in comparison with the exocrine activity of the pancreatic remnant (EAPR) calculated by the value of the product of volume of drained pancreatic juice and its amylase activity. The influences of those factors and other clinicopathologic data on PIA outcome were evaluated. RESULTS: The histological grade of fibrosis in the pancreatic stump was inversely correlated with EAPR (rs=0.5848, p=0.0011). Three patients had major leakages and 6 had minor leakages of PIA; all the patients with leakages had biliary or ampullary diseases, but not pancreatic diseases. The incidence of leakage of PIA was significantly higher in the patients with high values of EAPR (p<0.05). None with EAPR less than 10(7) had PIA leakage. The incidence of PIA leakage in low-grade fibrosis of the pancreatic stump was significantly higher than that in high-grade fibrosis (p<0.05). Other clinicopathologic data did not influence the incidence of leakage of PIA. CONCLUSIONS: The degree of fibrosis in the pancreatic stump is significantly related to the EAPR and affects the development of PIA insufficiency as a local factor. PMID- 12646988 TI - Plasma immunoadsorption therapy for Guillain-Barre syndrome: critical day for initiation. AB - Immunoadsorption plasmapheresis (IAPP) is a method of removing circulating immune factors that is used to treat Guillain-Barre syndrome (GBS). We retrospectively analyzed the data on our GBS patients. In 21 patients treated with IAPP, linear regression analysis showed that the time from the onset of symptoms to the initiation of IAPP was correlated with the time required for improvement by one Hughes functional grade. We investigated the critical day for initiating treatment, which we defined as the day when initiation of IAPP was significantly more likely to improve function by at least one Hughes grade when compared with the outcome in patients receiving supportive therapy (non-IAPP group). The critical day was found to be day 6 after the onset of GBS. PMID- 12646989 TI - [Histological changes elicited by hair removal lasers]. AB - Several different laser systems are currently used to remove unwanted hairs. In this study, we studied follicular changes following hair removal with ruby or alexandrite lasers at different fluences. Unwanted hairs were treated with a ruby laser (ICN, Photon Ics, UK) at 10, 14, 18 J/cm(2) or an alexandrite laser (Cynosure, USA) at 11, 14, 17 J/cm(2). A 3 mm punch biopsy was taken immediately after each laser exposure and one month later. Specimens were stained for histological observations. They were observed using immunohistochemistry to Factor VIII related antigen and PCNA, and also by the TUNEL method. Immediately after the laser exposure, moderate follicular damage was observed following treatment with either laser. One month later, cystic formation of hair follicles and foreign body giant cells were observed in skin treated with either laser. The similar influence of each laser treatment resulted in similar histological changes. In this study, the histological changes following treatment with a ruby or an alexandrite laser at the same fluence were considered to be similar. PMID- 12646990 TI - T1-weighted magnetic resonance imaging sequence appropriate for the evaluation of the longitudinal relaxation effect of superparamagnetic iron oxide: a phantom study. AB - The goal of this study was to determine a T1-weighted magnetic resonance (MR) imaging sequence appropriate for evaluating the longitudinal relaxation effect of superparamagnetic iron oxide (ferumoxides) in a phantom study. An agarose phantom that included various concentrations of ferumoxides (0 - 0.5 mmol/l in 0.05 mmol/l increments) was examined for six types of T1-weighted imaging sequences using a 1.5-T MR unit. Three-dimensional (3D) fast spoiled gradient-echo (SPGR) imaging with a short echo time showed a strong linear correlation between the concentration of ferumoxides and the enhancement ratio. Two-dimensional (2D) fast SPGR imaging showed a high signal-to-noise ratio of the phantom even at low ferumoxides concentrations. These results suggest that 3D fast SPGR imaging is an appropriate technique for the evaluation of the longitudinal relaxation effect of ferumoxides, and that 2D fast SPGR imaging can be useful for evaluating the longitudinal relaxation effect at lower ferumoxides concentrations. PMID- 12646991 TI - [A rare case of the lumbar ectopic kidney in Japanese adult]. AB - In this paper, about the left lumber ectopic kidney of a 87-years-old Japanese woman, we observed the size of her kidney, the form of her renal hilum, and the form of her calicopelvic system. In addition, we measured the index of cortex and medulla (C/M), and examined the difference between the lumber ectopic and the normal kidney. There was no significant difference in the size (length, width, thickness, weight), without the external form was the distorted oval of lumber ectopic kidney. The renal hilum was divided into the upper and lower parts by the septum consisted of the parenchyma of kidney. In the calicopelvic system of the left lumber ectopic kidney, an obstructive distension due to the oppression by branch of the renal vein was observed in the superior minor renal calices. It seemed that it was Fraley's syndrome. From the viewpoint of C/M in the lumber ectopic kidney, it was suggested that there was no difference in the renal function, because the C/M was within a normal range. PMID- 12646992 TI - [Diagnosis and treatment of lumbar spinal canal stenosis]. AB - Lumbar spinal canal stenosis (LSCS) was first described in 1954 by Verbiest, followed by the currently accepted international classification of LSCS in 1976 by Arnoldi. Briefly, LSCS is a nervous system syndrome that is characterized by neural symptoms in the lower extremities due to tightened cauda equina and spinal nerve root involvement. LSCS international classification consists of: (1) degenerative, (2) congenital developmental, (3) combined, (4) spondylolytic spondylolisthesis, (5) iatrogenic and (6) post traumatic stenosis. Degenerative stenosis-the most common type of LSCS-is caused by disc degeneration, osteoarthritis of the facet joint and hypertrophy of the ligamentum flavum. LSCS may also be the result of intervertebral disc degeneration, protruded intervertebral disc and/or bony spur compress cauda equina and spinal nerve root anteriorally, while degenerated facet joint and hypertrophied the ligamentum flavum compress cauda equina and spinal nerve root posteriorally? Most often, spondylolytic spondylolisthesis occurs at the fourth lumbar vertebrae in middle aged women. As a result of a slipping forward of the vertebra, cauda equina and spinal nerve roots can be tightened between the edge behind the top of lower vertebra and frontal edge of the lower part of upper lamina. Typical clinical symptoms of LSCS are low back pain, leg pain and intermittent claudication. Low back pain is chronic with secondary radiating pain in the buttock. The leg pain is called "sciatica", which tends to appear on the back of thigh, in the lateral aspect of lower leg and calf muscles, and which intensifies when the patient is fatigued. Intermittent claudication is a symptom associated with this syndrome. Often, patients with LSCS find it impossible to walk because of increased numbness and pain in their leg. Many patients report that after squatting for a few minutes they are able to resume walking. LSCS patients may also report dysaesthesia in the perineum area, and may also report urinary dysfunction ranging from extreme urgency to urinary delay. Patients who present with symptoms of LSCS should be seen by an orthopedic surgeon. Correct diagnosis by imaging and clinical examination, with appropriate conservative or operative treatment in a timely fashion should be encouraged in order to prevent irreversible nerve damage. PMID- 12646993 TI - [Mechanism of intractable low back pain and neural blockade]. AB - Low back pain is one of the most prevalent complaints in clinical medicine. Sensations from the axial skeleton and the surrounding tissues are only vaguely somatotopic and are non-specific in quality. The diagnosis of the mechanism or source of low back pain is therefore very challenging. The treatment of low back pain that recurs, persists or intensifies is also formidable, because there has been no evidence of various therapies for chronic back pain. From the prophylactic viewpoint of chronic pain, the most considerable matter is early elimination of severe pain under certain diagnosis. In this article, the mechanisms of passing into the chronic state, especially the development of neuropathic pain, and the utilities of diagnostic and therapeutic neural blockade for low back pain are discussed. PMID- 12646994 TI - [Acute suppurative thyroiditis in 7 year-old girl with piriform sinus fistula]. AB - A 7 year-old girl was admitted to our hospital with high grade fever and redness, swelling and tenderness in left neck. CT scan revealed cyst formation (4.5x3 cm) in left lobe of thyroid with swelling of surrounding lymphonodes. We diagnosed her as acute suppurative thyroiditis and treated her with intravenous antibiotics infusion and incisional drainage. After the treatment, the clinical course was uneventful. Pharyngograph revealed left piriform sinus fistula. PMID- 12646995 TI - Estimate of oxygen consumption and intracellular zinc concentration of human spermatozoa in relation to motility. AB - AIM: To investigate the human sperm oxygen/energy consumption and zinc content in relation to motility. METHODS: In washed spermatozoa from 67 ejaculates, the oxygen consumption was determined. Following calculation of the total oxygen consumed by the Ideal Gas Law, the energy consumption of spermatozoa was calculated. In addition, the zinc content of the sperm was determined using an atomic absorption spectrometer. The resulting data were correlated to the vitality and motility. RESULTS: The oxygen consumption averaged 0.24 micromol/10(6) sperm x 24h, 0.28 micromol/10(6) live sperm x 24h and 0.85 micromol/10(6) live motile sperm x 24h. Further calculations revealed that sperm motility was the most energy consuming process (164.31 mJ/10(6) motile spermatozoa x 24h), while the oxygen consumption of the total spermatozoa was 46.06 mJ/10(6) spermatozoa x 24h. The correlation of the oxygen/energy consumption and zinc content with motility showed significant negative correlations (r= -0.759; P<0.0001 and r=-0.441; P<0.0001, respectively). However, when correlating sperm energy consumption with the zinc content, a significant positive relation (r=0.323; P=0.01) was observed. CONCLUSION: Poorly motile sperm are actually wasting the available energy. Moreover, our data clearly support the "Geometric Clutch Model" of the axoneme function and demonstrate the importance of the outer dense fibers for the generation of sperm motility, especially progressive motility. PMID- 12646996 TI - Androgen-independent growth in LNCaP cell lines and steroid uridine diphosphate glucuronosyltransferase expression. AB - AIM: To investigate the mechanism of androgen-independent growth of prostate cancer after androgen ablation in LNCaP cells and the effect of glucuronidation activity. METHODS: To establish androgen-independent growth in prostate cancer LNCaP-SF, continuous passage was performed in androgen-stripped medium and the cells were evaluated for glucuronidation activity. The expression vector of antisense uridine diphosphate glucuronosyl-transferase (UGT) 2B15 cDNA was also constructed and evaluated. RESULTS: LNCaP-SF lead to a higher expression in UGT2B15 and their glucuronidation activity is 2.5 times higher than that of LNCaP cells. Significantly fewer LNCaP and LNCaP-SF than control were transfected with the antisense UGT2B15 cDNA, suggesting that UGT2B15 plays an important part in the glucuronidation activity of androgens in both cells. CONCLUSION: The alteration of UGT2B15 expression in LNCaP-SF cells is proposed as a biological characteristic involved in the growth of hormone-refractory prostate cancer. PMID- 12646997 TI - Effects of icariin on cGMP-specific PDE5 and cAMP-specific PDE4 activities. AB - AIM: To clarify the mechanism of the therapeutic action of icariin on erectile dysfunction (ED). METHODS: PDE5 was isolated from the human platelet and PDE4 from the rat liver tissue using the FPLC system (Pharmacia, Milton Keynes, UK) and the Mono Q column. The inhibitory effects of icariin on PDE5 and PDE4 activities were investigated by the two-step radioisotope procedure with [(3)H] cGMP/[(3)H]-cAMP. Papaverine served as the control drug. RESULTS: Icariin and papaverine showed dose-dependent inhibitory effects on PDE5 and PDE4 activities. The IC(50) of Icariin and papaverine on PDE5 were 0.432 micromol/L and 0.680 micromol/L, respectively and those on PDE4, 73.50 micromol/L and 3.07 micromol/L, respectively. The potencies of selectivity of icariin and papaverine on PDE5 (PDE4/PDE5 of IC(50)) were 167.67 times and 4.54 times, respectively. CONCLUSION: Icariin is a cGMP-specific PDE5 inhibitor that may be developed into an oral effective agent for the treatment of ED. PMID- 12646998 TI - Androgen and prostatic stroma. AB - AIM: To investigate the effect of androgen on the proliferation, differentiation and regression of canine prostatic stromal cells in vivo and human stromal cells in vitro. METHODS: Twenty-two dogs, including 15 normal prostate dogs and 7 prostatic hyperplasia dogs, had their serum concentration of testosterone and estrodiol determined by radioimmunoassay before and after castration. The expression of androgen receptor (AR) and estrogen receptor (ER) in the prostate were analysed by immunohistochemistry and semi-quantitative RT-PCR before and after castration. Light microscopy, transmission electron microscopy and TUNEL assay were carried out successively before and after castration to evaluate the prostatic histomorphology. In vitro serum-free cell cultures from human prostatic stroma were established and exposed to dihydrotestosterone (DHT). The proliferation of the cell culture was detected by MTT assay. The expression of TGFbgr, bFGF, AR, and smooth muscle cell (SMC) specific proteins (myosin and/or smoothelin) were detected using immunohistochemistry and RT-PCR. The differentiation from fibroblasts to smooth muscle cells was deduced by measuring the expression of SMC specific proteins. RESULTS: Before castration, the serum concentrations of testosterone and estrodiol were not statistically different between normal and hyperplasia groups. Following castration, the serum concentration of testosterone decreased rapidly in 2 days, and the concentration of estrodiol had no significant change compared with the pre-castration data. In the prostate, AR was presented in both the epithelial and stromal cells and the AR mRNA level was higher in hyperplasia than in normal prostate tissues (P<0.05). While ER predominantly existed in the prostate stromal cells and the ER mRNA had no difference between the hyperplasia and the normal group. Within the early phase of castration (0.05). Elastic fibers in the tunica albuginea of Group A were very rich and arranged regularly and undulatedly, but in Group B, most of the elastic fibers were replaced by collagenous fibers. In Group C, the tunica albuginea was mainly composed of thick and irregular-arranged collagenous fibers. In Group A, there were abundant smooth muscle fibers in the trabeculae of corpus cavernosum, but they were much less in Group C and scarce or even disappeared in Group B. In Groups B and C, the diminished/disappeared smooth muscle fibers were replaced by irregularly arranged collagenous fibers. CONCLUSION: In rats, androgen is essential for maintaining the normal structure of penile tunica albuginea and corpus cavernosum. PMID- 12647001 TI - Ultrastructural effect of sildenafil citrate on corpus cavernosum and other genital organs in female rats. AB - AIM: To determine the ultrastructural effects of sildenafil on the female genital organs. METHODS: Twenty female cycling Wistar albino rats weighing 250+/-20 g were randomly divided into two groups of 10 each. Rats of one group were gavaged with 0.5 mg.kg(-1).d(-1) of sildenafil 3 days in a week for 4 weeks and the other served as the controls. After cessation of treatment animals were sacrificed by cervical dislocation under methoxyflurane anaesthesia. The clitoris, vagina, uterus and bartholin glands were taken at the estrous and were fixed with 10% formalin solution for light microscopy and 2.5% glutaraldehyde and osmic acid for electron microscopy. RESULTS: Under the light microscope, the fibrocollageous tissue was found increased, the capillaries enlarged and the connecting tissue elements increased in the corpus cavernosum in the treated group. On electron microscopy, increased connective tissue, fibroblasts with notched nucleus, shorten immature collagen fibers without striation were seen. Abundant foldings and penetration with collagen bundles were observed in the basal membrane. Large connection complexes, especially gap junctions among the wide capillary endothelial cells were observed. CONCLUSION: There are evident histological changes due to sildenafil citrate in female rat corpus cavernosum. The clitoris and bartholin glands were the most effected organs. While the histopathological changes of clitoral tissue could be expected, an increase in the mass of bartholin gland was surprising. PMID- 12647002 TI - Cryopreservation-induced decrease in heat-shock protein 90 in human spermatozoa and its mechanism. AB - AIM: To study the protein changes of spermatozoa associated with sperm motility during sperm cryopreservation and its mechanism. METHODS: In 18 healthy men, the seminal sperm motility and HSP90 levels were studied before and after cryopreservation using SDS-PAGE, Western blotting and computerized image analysis. RESULTS: The sperm motility declined significantly after cryopreservation (P<0.01). The average grey level and the integrated grey level of sperm HSP90 before cooling were 34.1+/-3.2 and 243.0+/-21.6, respectively, while those after thawing were 23.2+/-2.5 and 105.7+/-28.5, respectively. Both parameters were decreased significantly (P<0.01). No HSP90 was found in the seminal plasma before and after cryopreservation. CONCLUSION: HSP90 in human spermatozoa was decreased substantially after cryopreservation. This may result from protein degradation, rather than leakage into the seminal plasma. PMID- 12647003 TI - Chlamydia trachomatis and sperm lipid peroxidation in infertile men. AB - AIM: To relate the presence of anti-Chlamydial trachomatis IgA in semen with sperm lipid membrane peroxidation and changes in seminal parameters. METHODS: Semen samples of the male partners of 52 couples assessed for undiagnosed infertility were examined for the presence of IgA antibody against C. trachomatis. The level of sperm membrane lipid peroxidation was estimated by determining the malondialdehyde (MDA) formation. RESULTS: Sperm membrane of infertile males with positive IgA antibodies against C. trachomatis showed a higher level of lipid peroxidation than that of infertile males with negative IgA antibody (P<0.05). There was a positive correlation (P<0.01) between the level of C. trachomatis antibody and the magnitude of sperm membrane lipid peroxidation. All the other tested semen parameters were found to be similar in the two groups. CONCLUSION: The activation of immune system by C. trachomatis may promote lipid peroxidation of the sperm membrane. This could be the way by which C. trachomatis affects fertility. PMID- 12647004 TI - Expression and localization of Smad1, Smad2 and Smad4 proteins in rat testis during postnatal development. AB - AIM: To study the expression and regulation of Smad1, Smad2 and Smad4 proteins (intracellular signaling molecules of transforming growth factor-b family) in rat testis during postnatal development. METHODS: The whole testes were collected from SD rats aged 3, 7, 14, 28 and 90 (adult) days. The cellular localization and developmental changes were examined by immunohistochemistry ABC method with the glucose oxidase-DAB-nickel enhancement technique. Quantitative analysis of the immunostaining was made by the image analysis system. The Smads proteins coexistence in the adult rat testis was tested by the double immune staining for CD14-Smad4 and Smad2-Smad4. The protein expression of Smad during rat testicular development was examined by means of Western blots. RESULTS: Smad1, Smad2 and Smad4 were present throughout testicular development. The immunostaining of Smad1 and Smad2 were present in spermatogenic cells. A positive immunoreactivity was located at the cytoplasm, but the nucleus was negative. Smad1 was immunolocalized at the d14, d28 and adult testes, while Smad2, at the d7, d14, d28 and adult testis. There was positive immunoreaction in the Sertoli cells and Leydig cells as well. The immunolocalization of Smad4 was exclusively at the cytoplasm of Leydig cells and the nuclei were negative throughout the testicular development. No expression was detected in the germ cells. The results of image and statistical analysis showed that generally the expression of Smad1, Smad2 and Smad4 in the testis tended to increase gradually with the growth of the rat. CONCLUSION: The present data provide direct evidences for the molecular mechanism of TGF-bgr action in rat testes during postnatal development and spermatogenesis. PMID- 12647005 TI - Prosaposin ablation inactivates the MAPK and Akt signaling pathways and interferes with the development of the prostate gland. AB - The recent development of a prosaposin -/- mouse model has allowed the investigation of the role of prosaposin in the development of the male reproductive organs. A morphometric analysis of the male reproductive system of 37 days old mice revealed that prosaposin ablation produced a 30 % reduction in size and weight of the testes, 37 % of the epididymis, 75 % of the seminal vesicles and 60 % of the prostate glands. Light microscopy (LM) showed that smaller testis size from homozygous mutant mice was associated with reduced spermiogenesis. Both, dorsal and ventral lobules of the prostate glands were underdeveloped in the homozygous mutant. LM analysis also showed that prostatic alveoli were considerably smaller and lined by shorter epithelial cells in the homozygous mutant. Smaller tubular diameter and shorter undifferentiated epithelial cells were also observed in seminal vesicles and epididymis. In the efferent ducts of the homozygous mutant mice, the epithelium was composed exclusively of ciliated cells in contrast to the heterozygotes, which showed the presence of nonciliated cells. Radioimmunoassays demonstrated that testosterone levels were normal or higher in mice with the inactivated prosaposin gene. Immunostaining of prostate sections with an anti-androgen receptor antibody showed that the epithelial cells lining the alveoli express androgen receptor in both the heterozygous and homozygous tissue. Similarly, sections immunostained with antibodies to the phosphorylated MAPKs and Akts strongly reacted with tall prostatic secretory cells in prostate from heterozygous mouse. On the other hand, the epithelial cells in the homozygous prostate remained unstained or weakly stained. These findings demonstrate that inactivation of the prosaposin gene affected the development of the prostate gland and some components of the MAPK pathway. PMID- 12647006 TI - Intratunical bupivacaine and methylprednisolone instillation for scrotal pain after testicular sperm retrieval procedures. AB - AIM: To investigate the effect of intratunical instillation of bupivacaine and methylprednisolone for scrotal pain, swelling and peritesticular fibrosis due to testicular sperm retrieval procedures. METHODS: A total of 65 patients were randomly divided into two groups. In the instillation group (GI), 34 patients were administered 2.5 mL of 0.5 % bupivacaine combined with 10 mg/ml methylprednisolone before closure of the tunica vaginalis. In the control group (GC), 31 patients only received analgesics postoperatively by intramuscular route. The pain (by visual analogue scale, VAS) and duration of pain-free period after surgery between the two groups were evaluated at 2 and 4 h and at days 2 and 7 postoperatively. RESULTS: The mean pain scores were significantly lower in the GI than in the GC group at 2 and 4 h after surgery (P<0.05 and P<0.01, respectively). The mean duration of pain free interval after the procedure was 47.8 +/-16.9 (12-72) h in GI, which was significantly longer than that in GC [(9.9 +/-3.6; 4-20) h]. Besides, in the GI, 29 % of patients were completely free from pain and 67 % had no scrotal swelling, but in the GC, all the patients required additional NSAID injection due to pain and only 3 % had no scrotal swelling. CONCLUSION: This study confirms that direct intratunical instillation of bupivacaine and methylprednisolone around the testis reduces the postoperative pain, scrotal swelling and peritesticular fibrosis. PMID- 12647007 TI - Intracytoplasmic sperm injection in cases with history of in vitro fertilization failure. AB - AIM: To evaluate the effect of intracytoplasmic sperm injection (ICSI) in the management of cases with a history of conventional in vitro fertilization (IVF) failure. METHODS: Two groups of patients, 19 with normal semen parameters and a history of IVF failure (metaphase II oocytes: 0-30 %) and 28 with severe male factor infertility received ICSI technology during the same period. Ovarian stimulation was achieved by conventional procedure. Transvaginal ultrasound guided oocyte collection was done 35-37 h after human chorionic gonadotrophin (hCG) injection. Only metaphase II oocytes were selected for microinjection. RESULTS: Fertilization was achieved with ICSI in all the patients. The fertilization rate (75.6 % +/-21.1 % vs. 73.9 % +/-19.2 %), cleavage rate (85.1 % +/-19.3 % vs. 82.7 % +/-22.1 %), clinical pregnancy rate per embryo transfer cycle (31.6 % vs. 28.6 %) and implantation rate per embryo (15.3 % vs. 14.4 %) did not differ significantly between the two groups. CONCLUSION: ICSI is a valuable method for couples with a history of IVF failure. These patients may have a similar ICSI result as in severe male infertility. PMID- 12647008 TI - Successful pregnancy and birth after intrauterine insemination using caput epididymal sperm by percutaneous aspiration. AB - AIM: To manage male infertility with obstructive azoospermia by means of percutaneous epididymal sperm aspiration (PESA) and intrauterine insemination (IUI). METHODS: Ninety azoospermic patients with congenital bilateral absence of the vas deferens (BAVD, n=58) or bilateral caudal epididymal obstruction (BCEO, n=32) requesting for fine needle aspiration (FNA), PESA and IUI were recruited. The obstruction was diagnosed by vasography and determination of the fructose, carnitine and alpha-glucosidase levels in the seminal fluid. RESULTS: The mean sperm motility, density, abnormal sperm and total sperm count of the caput epdidymis were 16 %+/-22 %, (12+/-31) x 10(6)/mL, 55 %+/-36 % and (16+/-14) x 10(6), respectively. In the 90 couples, a total of 74 PESA procedures and 66 cycles of IUI were performed. Three pregnancies resulted, including one twin pregnancy giving birth to two healthy boys, one single pregnancy with a healthy girl and another single pregnancy aborted at week 6 of conception. The pregnancy rate per IUI cycle was 4.5 %. CONCLUSION: The birth of normal, healthy infants by IUI using PESA indicates that the caput epididymal sperm possess fertilization capacity. The PESA-IUI programme is a practical and economical procedure for the management of patients with obstructive azoospermia. PMID- 12647009 TI - First experience with intracytoplasmic sperm injection for extreme oligozoospermia associated with Crohn's disease and 6-mercaptopurine chemotherapy. AB - AIM: To describe the reproductive outcome following intracytoplasmic sperm injection (ICSI) for male factor infertility associated with Crohn's disease and 6-mercaptopurine (6-MP) chemotherapy. METHODS: The male partner of a couple suffered from severe Crohn's disease and received a 3-month course of 6-MP for this condition. Two spontaneous conceptions were established before 6-MP, although post-chemotherapy semen analysis found the sperm concentration to be 8,000/mL. In vitro fertilization (IVF) with ICSI and embryo transfer was performed. RESULTS: The woman underwent an uncomplicated controlled ovarian hyperstimulation sequence using a combined rec-FSH+hMG protocol, following late luteal phase pituitary downregulation. This culminated in the retrieval of 18 oocytes, 11 of which were fertilized with ICSI. She later delivered a normal male infant without urogenital anomaly. Four nontransferred blastocysts were cryopreserved. CONCLUSION: This report describes the first successful birth after ICSI for severe oligozoospermia associated with Crohn's disease and 6-MP therapy. We outline salient features of Crohn's disease, 6-MP pharmacology, and their relevance to human fertility. PMID- 12647010 TI - Effect of angiotensin II and angiotensin(1-7) on hematopoietic recovery after intravenous chemotherapy. AB - PURPOSE: Previous studies have shown that angiotensin peptides stimulate the proliferation of hematopoietic progenitors in vitro, promote survival after exposure to lethal irradiation as well as accelerate the recovery of white blood cells (WBC), i.e., lymphocytes, monocytes and neutrophils, and platelets. These changes in the level of formed elements in the blood after irradiation was thought to be due to increases in the numbers of bone marrow progenitors including myeloid, erythroid and megakaryocyte progenitors by the action of angiotensin peptides. In view of these findings, the effect of angiotensin peptides on recovery after chemotherapy was assessed. MATERIALS AND METHODS: The effect of angiotensin II (AII) and angiotensin(1-7) (A1-7) on the recovery of WBC and platelets in the blood, as well as the number of myeloid, erythroid and megakaryocyte progenitors in the bone marrow and the number of myeloid progenitors in the blood after intravenous administration of chemotherapeutic drugs was assessed in a mouse model. RESULTS. In initial studies, subcutaneous administration of 10 or 100 microg/kg per day of AII starting either 2 days before or 2 days after intravenous administration of 5-fluorouracil (5FU) accelerated WBC recovery (return to baseline between 7 and 14 days). Further, consistent with previous observations, the number of myeloid progenitors in the bone marrow and blood was increased after systemic administration of angiotensin peptides. The comparability of A(1-7) and AII in their effect on hematopoietic recovery after chemotherapy was shown in subsequent studies. Daily administration of both AII and A(1-7) increased platelet numbers in the peripheral blood and myeloid, erythroid and megakaryocyte progenitors in the bone marrow. As 5FU is not a stem cell toxin, these studies were repeated with administration of A(1-7) initiated before or after intravenous cyclophosphamide. Following treatment with A(1-7) before cyclophosphamide the numbers of circulating WBC initially increased and then decreased starting on day 14. Following treatment with A(1-7) 2 days after cyclophosphamide the numbers of WBC and the numbers of myeloid progenitors increased in the peripheral blood and bone marrow. CONCLUSIONS: These findings suggest that angiotensin peptides accelerate hematopoietic recovery in multiple cellular lineages after chemotherapy, perhaps through an increase in the number of early hematopoietic progenitors. PMID- 12647011 TI - A population pharmacokinetic model for doxorubicin and doxorubicinol in the presence of a novel MDR modulator, zosuquidar trihydrochloride (LY335979). AB - PURPOSE: To develop a population pharmacokinetic model for doxorubicin and doxorubicinol in the presence of zosuquidar.3HCl, a potent P-glycoprotein inhibitor. METHODS: The population approach was used (implemented with NONMEM) to analyse doxorubicin-doxorubicinol pharmacokinetic data from 40 patients who had received zosuquidar.3HCl and doxorubicin intravenously (separately in cycle 1 and concomitantly in cycle 2 over 48 h and 0.5 h, respectively). RESULTS: A five compartment pharmacokinetic model (including three compartments for doxorubicin pharmacokinetics with two pathways for doxorubicinol formation) best described the doxorubicin-doxorubicinol pharmacokinetics in the presence of zosuquidar.3HCl. Doxorubicin clearance (CL), peripheral volume of distribution (V2) and doxorubicinol apparent clearance (CLm/fm) and apparent volume of distribution (Vm/fm) were 62.3 l/h, 2360 l, 143 l/h and 3150 l, respectively, in the absence or presence of low doses of zosuquidar.3HCl (<500 mg). In the presence of high doses of zosuquidar.3HCl (>or=500 mg), these values decreased by 25%, 26%, 48% and 73%, respectively, and doxorubicinol pharmacokinetics were characterized by a delayed t(max) (24 h versus 4 h), which led to the inclusion of the parallel pathways. A decrease in the objective function ( P<0.005) was observed when the impact of zosuquidar.3HCl was accounted for. CONCLUSIONS: This integrated parent-metabolite population pharmacokinetic model accurately characterized the increase in doxorubicin and doxorubicinol exposure (1.33- and 2 fold, respectively) in the presence of zosuquidar.3HCl (>or=500 mg) and provided insights into the pharmacokinetic interaction, which may be useful in designing future clinical trials. PMID- 12647012 TI - Arsenic trioxide induces apoptosis equally in T lymphoblastoid leukemia MOLT-4 cells and P-gp-expressing daunorubicin-resistant MOLT-4 cells. AB - PURPOSE: To investigate the effects of arsenic trioxide (As(2)O(3)) on human T lymphoblastoid leukemia MOLT-4 cells and P-gp-expressing daunorubicin-resistant MOLT-4 (MOLT-4/DNR) cells. METHODS: Cell growth was measured by an MTT assay. Cell viability was determined by a dye exclusion test. The level of P-gp expression was estimated using phycoerythrin-conjugated anti-P-gp monoclonal antibody 17F9. The function of P-gp was evaluated in terms of rhodamine 123 (Rh123) efflux. The percentage of cells undergoing apoptosis was determined by flow cytometry after staining with annexin V-FITC and propidium iodide. RESULTS: As(2)O(3) inhibited the growth and survival of MOLT-4 and MOLT-4/DNR cells in a time- and dose-dependent manner. The 50% inhibitory concentrations of As(2)O(3) (IC(50)) against the growth of these cell lines were 5.1 micromol/l and 5.0 micromol/l, respectively, when the cells were treated with As(2)O(3) for 96 h. As(2)O(3) induced an apoptotic morphology in both MOLT-4 and MOLT-4/DNR cell lines. These effects of As(2)O(3) were time- and dose-dependent when the two cell lines were incubated in the presence of 1-8 micromol/l of As(2)O(3) for 3-144 h. As(2)O(3) treatment for 3 to 24 h at 5.0 micromol/l did not change the percentage of P-gp-expressing cells or the efflux ability of MOLT-4/DNR cells. CONCLUSION: As(2)O(3) inhibited growth and induced apoptosis equally in MOLT-4 and MOLT-4/DNR cells, and this suppressive effect did not influence P-gp expression or function in MOLT-4/DNR cells. PMID- 12647013 TI - Population pharmacokinetics of oxaliplatin. AB - The objective of this study was to explore correlations between a variety of covariates and oxaliplatin ultrafilterable and blood pharmacokinetic parameters. Data from 40 patients receiving oxaliplatin combined with 5-fluorouracil and levofolinic acid as standard treatment for advanced colorectal cancer were analysed. Plasma ultrafilterable, blood, and urine platinum concentrations were determined by flameless atomic absorption spectrophotometry. Data were analysed according to a population pharmacokinetic method using the NONMEM program. The best fit for oxaliplatin plasma ultrafilterable clearance (CL) was given by the following equation, which considers four covariates: body surface area (BSA, in metres squared), age (in years), sex (0 if male, 1 if female), and serum creatinine (Scr, in micromoles per liter): CL (l/h)=5.49xBSA+4.55xBSAx(140 AGE)x(1-0.15xSEX)/Scr. By taking into account these covariates, the interindividual variability in CL decreased from 43% to 33%. Renal clearance represented 34% of the overall elimination. This value was obtained by recovering urine over only 5 h from the beginning of the infusion and modelling the data using NONMEM. We would recommend the use of this methodology for pharmacokinetic studies in oncology in which renal clearances of the drug are presently rarely explored. The oxaliplatin blood concentrations versus time observed during the three-cycle period were well-described by a three-compartment model with first order elimination from the central compartment. No significant intrapatient pharmacokinetic variability was observed between cycles. The relationship we obtained using the population approach between oxaliplatin CL and covariates may allow rational reduction of oxaliplatin dose in cases of elevated serum creatinine levels. PMID- 12647014 TI - Inhibition of basolateral transport and cellular accumulation of cDDP and N acetyl- L-cysteine-cDDP by TEA and PAH in the renal proximal tubule. AB - PURPOSE: The objective of this study was to determine the effect of para aminohippurate (PAH) and tetraethylammonium (TEA) on basolateral cellular accumulation (C(Pt)) and bath-to-lumen transepithelial transport rates (J(B)(- >)(L)) of platinum from cisplatin (cDDP) and a conjugate of cDDP, N-acetyl- L cysteine-cDDP (NAC-cDDP), in S(1), S(2), and S(3) segments of the rabbit proximal tubule. METHODS: Cellular accumulations and transport rates were determined using the isolated perfused tubule technique and samples were analyzed by ICP-MS. RESULTS: First, to establish the control data, each tubular segment was bathed in free cDDP (2 m M) which resulted in no observable toxicity. Next, TEA (4 m M) was added to the bathing solution containing cDDP. This resulted in a reduction in platinum J(B)(-->)(L) by approximately 75% in the S(1) segment and 50% in the S(2) and S(3) segments. C(Pt) was reduced by 80-90% in relation to control values with no observable changes in toxicity. In the next experiment, exposure of the basolateral membrane to NAC-cDDP (2 m M) elicited pronounced toxicity after 20-30 min of perfusion. The J(B)(-->)(L) for NAC-cDDP was similar for each of the three nephron segments. There were no significant differences in the ability of these three segments to accumulate NAC-cDDP, but the conjugate increased uptake of platinum by 200-300% in the S(1) and S(2) segments, with no significant change in the S(3) segments, compared cDDP control values. The presence of PAH (4 m M) in the bathing solution significantly reduced J(B)(-->)(L) (by approximately 90%) for NAC-cDDP in all segments and the C(Pt) by approximately 80%. This also abrogated the NAC-cDDP-induced toxicity. CONCLUSIONS: There was axial heterogeneity among the basolateral membranes of the S(1), S(2), and S(3) segments of the proximal tubule in accumulating free cDDP and transport of NAC cDDP. Generally, the NAC-cDDP molecule was transported more avidly than free cDDP across the basolateral membrane, except in the S(3) segment, where accumulation was similar to that of free cDDP. It is concluded that a PAH-sensitive organic anion transporter is involved in the accumulation of NAC-cDDP at the basolateral membrane and a TEA-sensitive organic cation transport system is involved in the accumulation of free cDDP. PMID- 12647015 TI - Effect of dexrazoxane on doxorubicin pharmacokinetics in young and old rats. AB - PURPOSE: Although dexrazoxane (ICRF-187) is used clinically to protect against doxorubicin cardiotoxicity, the age-related effect of dexrazoxane on doxorubicin pharmacokinetics has not been well studied. METHODS: We therefore examined the effect of pretreatment with dexrazoxane (50 mg kg(-1) i.p. 1 h prior to administration of doxorubicin 2 mg kg(-1) i.v. bolus) on doxorubicin and doxorubicinol pharmacokinetics in Fischer 344 rats at 5 months of age (young adult) and 22 months of age (old). RESULTS: Dexrazoxane had no major effects on doxorubicin or doxorubicinol pharmacokinetics in plasma or heart in either young or old rats. However, age had significant effects on anthracycline pharmacokinetics. Early plasma concentrations were increased and systemic clearance of doxorubicin was decreased in old compared with young rats. Cardiac concentrations of doxorubicin (AUC) were significantly increased in old rats. In addition cardiac doxorubicinol concentrations (AUC 0-72 h) were increased by over 80% in old compared to young rats. CONCLUSION: The results suggest that dexrazoxane does not alter doxorubicin pharmacokinetics. In contrast, aging in the rat model is associated with altered doxorubicin and doxorubicinol pharmacokinetics, in particular in the heart. These changes could increase the risk of anthracycline cardiotoxicity with age. PMID- 12647016 TI - Effects of chlorin-mediated photodynamic therapy combined with fluoropyrimidines in vitro and in a patient. AB - Although photodynamic therapy (PDT) is becoming an additional cancer therapy, only little is known about its interactions with other drugs and treatment modalities in vitro and in vivo. We investigated the combination of 5-fluoro-2' deoxyuridine (5FdUr), a chemotherapeutic drug, with 5,10,15,20-tetra( m hydroxyphenyl)chlorin (mTHPC), a potent photosensitizer. Two cell lines, MCF-7 and LNCaP, were either simultaneously or consecutively incubated with both drugs and irradiated with laser light to activate mTHPC, and cell survival was determined. The combination of the two treatments was additive or antagonistic in LNCaP cells but additive or synergistic in MCF-7 cells depending on the protocol and concentration of 5FdUr. In one patient with multiple basal cell carcinoma, the effect of the combination of 5-fluorouracil administration followed by PDT resulted in significantly stronger effects than expected, leading to severe oedema, redness and ulceration. The healing process was delayed by 2 months compared to PDT alone. It is therefore important to find optimal conditions under which PDT and chemotherapy combinations do not fall outside the therapeutic window. PMID- 12647017 TI - Correlation between clinicopathologic factors and kinetics of metabolic enzymes for 5-fluorouracil given to patients with colon carcinoma by two different dosage regimens. AB - PURPOSE: Using tumor tissue specimens from colon carcinoma patients given 5 fluorouracil (5-FU) by two different administration methods, we investigated the degree of correlation between clinicopathologic factors and the kinetics of metabolic enzymes for 5-FU. METHODS: Group A patients received 500 mg/day of 5-FU as a rapid infusion over 2 h for 3 days preoperatively, and group B patients received 500 mg/day of 5-FU as a continuous infusion for 3 days preoperatively. The activities of orotate phosphoribosyl transferase (OPRT), thymidine phosphorylase (TP), uridine phosphorylase (UP), and dihydropyrimidine dehydrogenase (DPD) were measured by separation on ion-exchange filter paper, by high-performance liquid chromatography (HPLC) with UV detection and by HPLC with radioactive flow monitoring, respectively, and the [(3)H]-5-fluorodeoxyuridine monophosphate binding site in thymidylate synthetase (TS) was determined as an index of the amount of TS using a radiobinding assay. The TS inhibition rate (TSIR) was calculated from the formula: 1-(TS(free)/TS(total))x100. Finally, 5-FU incorporation into RNA (FRNA) was measured by capillary gas chromatography-mass spectrometry. RESULTS: In group A patients, FRNA showed a positive correlation only with OPRT, and TSIR showed positive correlations with OPRT and TP, but a negative correlation with TS. In group B patients, FRNA showed a negative correlation with DPD, and TSIR showed a negative correlation with TS. No difference in TSIR levels was seen between groups A and B. FRNA was higher in group A than in group B, but the difference was not statistically significant. CONCLUSION: The method of administration may influence 5-FU metabolism in colon carcinoma patients. PMID- 12647018 TI - Modulation of multidrug resistance-associated protein 1 (MRP1) by p53 mutant in Saos-2 cells. AB - The p53 tumor suppressor gene is one of the most frequently mutated genes in human cancer and the mutation is correlated with a poor prognosis in cancer therapy. Upregulation of multidrug resistance-associated protein 1 (MRP1) and increase in drug resistance have been found to be induced by p53 mutation. Human osteosarcoma Saos-2 cells, a p53-null cell line, was transfected with p53 with mutations at codon 143 (V to A), 175 (R to H), 248 (R to W), 273 (R to H) and 281 (D to G). Among the different transfectants, overexpression (about 42-fold) of MRP1 was detected in p53-R175H cells. Furthermore, the p53-R175H cells were 2.5 fold more resistant to doxorubicin (DOX) and had a 4-fold greater DOX efflux rate than the control cells 1 h after DOX treatment. Transfection with antisense MRP1 oligonucleotides demonstrated a DOX sensitization effect (about 2-fold) in p53 R175H transfectants but not in control cells. In addition, transfection with antisense p53 oligonucleotides greatly suppressed MRP1 expression and reversed DOX resistance in p53-R175H cells but had no effect in control cells. The results suggested that p53-R175H might induce MRP1 expression and DOX resistance in cells. PMID- 12647019 TI - Extensive hepatic replacement due to liver metastases has no effect on 5 fluorouracil pharmacokinetics. AB - PURPOSE: The influence of liver metastases on the pharmacokinetics of 5 fluorouracil (5-FU) and its metabolite 5,6-dihydrofluorouracil (DHFU) was studied in patients with liver metastases from gastrointestinal cancer ( n=16) and compared with a control group of patients with nonmetastatic gastrointestinal cancer ( n=18). METHODS: Patients were assigned to two different groups based on the presence of liver metastases. The percentage of hepatic replacement was determined with CT and ultrasonography and classified as <25%, 25-50% or >50% of the total liver volume. Chemotherapy consisted of leucovorin 20 mg/m(2) per day plus 5-FU 425 mg/m(2) per day, both for 5 days. Blood sampling was carried out on the first day of the first chemotherapy cycle. 5-FU and DHFU were quantified in plasma by HPLC. A four-compartment parent drug-metabolite model with nonlinear Michaelis-Menten elimination from the central compartment of the parent drug (5 FU) was applied to describe 5-FU and DHFU pharmacokinetics. RESULTS: No effect of liver metastases on 5-FU clearance was observed. The effects of 18 covariables on pharmacokinetic parameters were also studied in a univariate correlation analysis. Body surface area was positively correlated with the distribution volume of 5-FU in the central compartment and with V(max) ( r=0.65 and r=0.54, respectively). CONCLUSIONS: There is no need for dose adjustment of 5-FU as a standard procedure in patients with liver metastases and mild to moderate elevations in liver function tests. PMID- 12647020 TI - Sonodynamic therapy with photofrin II on AH130 solid tumor. Pharmacokinetics, tissue distribution and sonodynamic antitumoral efficacy of photofrin II. AB - BACKGROUND: The pharmacokinetics and tissue distribution of photofrin II (PF) and its efficacy in sonodynamic therapy were studied in rats bearing AH130 solid tumors. MATERIALS AND METHODS: In order to find the optimum timing of the ultrasound exposure after administration of PF, the PF concentrations in plasma, skin, muscle and tumor were measured and pharmacokinetically analyzed. Antitumor effects were estimated by measuring tumor size. RESULTS: Since the highest concentration of PF in tumors occurred 24 h after administration, ultrasound administration 24 h after the intravenous administration of PF was chosen. Ultrasound alone showed a slight antitumor effect, which became increasingly significant as the dose of PF was increased, while PF alone showed no significant effect. CONCLUSIONS: PF significantly sensitized solid tumors to the antitumor effect of ultrasound in a synergistic manner. PMID- 12647022 TI - Vasa vasorum of the human great saphenous vein. AB - The distribution of the vasa vasorum of the human great saphenous vein (GSV) was studied on veins taken both post-mortem and peroperatively. It was found that the stems of feeding vessels approach the venous wall at intervals of 1.5-2.5 cm; their smaller branches first passed the fascial compartments of the GSV and then entered the adventitia at intervals of 0.5-1.5 cm on both the stem and the largest tributaries of the GSV. In the stem regions vasa vasorum arteries and veins ran together but, between neighboring stems, isolated venae vasorum were regularly found which opened individually into terminal segments of the largest tributaries of the GSV. Neither by dissection nor by injection methods were venae vasorum found to open directly into the lumen of the GSV stem. The total thickness of the media ranged between 500 and 1300 micro m, according to the state of constriction of the venous wall before fixation. Two structurally different layers of GSV tunica media were present: an inner loose layer and an outer dense layer, both of similar thickness. The innermost capillaries of the vasa vasorum network were found in all cases on the border between the two layers of media. No lymphatic was found in any of the layers of GSV wall. From the findings the authors recommend extremely careful dissection of the GSV wall during in situ grafting surgery, to ensure the best viability of the venous wall. PMID- 12647021 TI - Vinorelbine and docetaxel as first-line chemotherapy in metastatic breast cancer. AB - PURPOSE: To evaluate the efficacy and tolerability of a combination of vinorelbine (VNR) and docetaxel (DOC) as first-line chemotherapy in patients with metastatic breast cancer. PATIENTS AND METHODS: The study group comprised 40 women with untreated metastatic breast cancer with visceral (85%) and bone (70%) metastases. Of the 40 patients, 24 (60%) had previously received adjuvant chemotherapy, which had included anthracyclines in 12 patients (30%). Treatment consisted of VNR 25 mg/m(2) on days 1 and 5, and DOC 75 mg/m(2) on day 1 every 3 weeks. Depending on the neutrophil nadir (grade 3 or 4 neutropenia by WHO criteria) recombinant human granulocyte colony-stimulating factor (G-CSF) 5 micro g/kg on days 2-4 and 6-13 was given for all subsequent treatment cycles. RESULTS: The overall response rate (ORR) was 40% (95% confidence interval, CI 15-65). Six patients (15%) achieved a complete response (CR) and ten patients (25%) achieved a partial response (PR). Stable disease (SD) was observed in six patients (15%), and 18 patients (45%) had progressive disease (PD). The median duration of response was 8 months and the median predictive time to progression (TTP) was 6 months. The main toxicity was neutropenia grade 3 and 4 in 28 patients (70%). Febrile neutropenia requiring hospitalization occurred in 12 patients (30%). Grade 3 or 4 anemia was seen in two patients (5%) and grade 3 or 4 thrombocytopenia was seen in one patient (2.5%). Severe nonhematologic toxicity, except alopecia, was uncommon and included stomatitis in two patients (5%), vomiting in two (5%) and diarrhea in one (2.5%). There were no treatment-related deaths. CONCLUSIONS: The combination of VNR and DOC at the doses used in this study showed moderate activity as first-line chemotherapy in metastatic breast cancer. Neutropenia was considerable despite G-CSF administration. PMID- 12647023 TI - The supra-auricular arterial network: anatomical bases for the use of superior pedicle retro-auricular skin flaps. AB - The authors present an anatomical study on vascularization of the retro-auricular skin, focusing on the origin, distribution and anastomoses of the superior auricular artery. This artery is used as the pedicle of a retro-auricular flap to repair defects in the upper third of the auricle and in the temporal region. The study was carried out on 13 fresh cadavers, with a total of 25 auricles. The common carotid artery had previously been injected with dyed latex. The superior auricular artery was found in all specimens. It had constant course and caliber, mean axial length 2.4 cm, mean caliber 0.8 mm. This artery connected the superficial temporal artery, or its parietal branch, and the posterior auricular artery network. This branch proved a reliable vascular pedicle for the mobilization of retro-auricular flaps. The results of this study are presented together with preliminary clinical results obtained using an antero-superior retro-auricular flap, never previously described, to reconstruct the superior third of the auricle and the temporal region. PMID- 12647024 TI - Radio-anatomic study of the pancreatic duct by MR cholangiopancreatography. AB - To compare the performance of MR-cholangiopancreatography (MRCP) and that of classical anatomy in the depiction of the main pancreatic duct, 50 MRCP examinations were done in patients free of pancreatic disease. Axial and coronal sections 20 mm thick were obtained in a Single Shot Fast Spin Echo (SSFSE) sequence. The following were analyzed: (1) visibility of pancreatic duct structures, (2) form of the main pancreatic duct, (3) various angulations of the duct and (4) diameter of the duct. Anatomic variants were noted. These findings were compared with anatomic and radio-anatomic (ERCP) data in the literature. The main pancreatic duct was visualized in 100% of cases and the accessory pancreatic duct in 61%. The form, diameter and angulations of the various segments of the pancreatic duct were similar to those reported in the literature. These findings are reported in the axial and coronal planes. Comparison with major anatomic classifications was not possible. MRCP enables in vivo anatomic exploration of the main pancreatic duct. Horizontal sections provided new radio-anatomic information. The technique nevertheless remains limited by poor spatial resolution. PMID- 12647025 TI - Role of the sinu-vertebral nerve in low back pain and anatomical basis of therapeutic implications. AB - Low back pain is frequent and results in major disability for patients. This anatomical study was done to understand mechanisms involved in that pain. Two kinds of innervation are present in the lumbar spine: one depends on the somatic nervous system and the other on the sympathetic nervous system. The sympathetic nerves are the sinu-vertebral nerves and the rami communicantes which innervate the intervertebral disc, the ventral surface of the dura mater, the longitudinal dorsal ligament and the longitudinal ventral ligament. The sinu-vertebral nerve was described first by Luschka in 1850. This nerve is implicated in diffuse low back pain because of its pathway and its sympathetic component. This nerve cannot directly reach a somatic element at each level of the lumbar spine, so must first reach the L2 spinal ganglion. Thus, there is a "hole" in the somatic innervation between L3 and L5 because the dorsal nerves do not reach the skin at these levels. The pain therefore takes another route through the sympathetic system. Discogenic pain is mediated by the sinu-vertebral nerves, and through the rami communicantes reaches the L2 spinal ganglion. Anatomical and clinical features reinforce this hypothesis. PMID- 12647026 TI - Can intestinal innervation be preserved in pancreatoduodenectomy for cancer? Results of an anatomical study. AB - Twenty dissections were carried out, in all of which the splanchnic nerves, celiac plexuses, capital pancreatic plexus and superior mesenteric plexus were identified and traced. The capital pancreatic plexus was formed from two bundles, the first taking its origin from the right celiac plexus, the second from the superior mesenteric plexus. These two bundles joined together just behind the head of the pancreas. Two preganglionic bundles, a ganglion and two postganglionic bundles composed the superior mesenteric plexus. Postganglionic bundles received fibers from both right and left celiac plexuses. In small cancers a thin layer of nervous tissue around the superior mesenteric artery might be spared in order to avoid diarrhea from intestinal denervation. This study has provided anatomical evidence that a part of the mesenteric plexus, which receives fibers from both left and right celiac plexuses, maintains a sufficient intestinal innervation. PMID- 12647027 TI - Does the meniscus exist in the elbow joint in children? AB - The authors report a case of meniscus at the elbow joint in a 15-month-old infant causing a limitation of elbow extension. Histological examination demonstrated that this tissue was not a synovial fold or a chondroid metaplasia of the synovial fold. As a meniscus does not appear at any stage of the embryological evolution of the elbow joint, it has been concluded that the presence of the meniscus can be considered as an abnormal condition. PMID- 12647028 TI - Combined transmission and emission tomography in a vastus intermedius muscle stress syndrome. PMID- 12647029 TI - TRPs make sense. AB - Drosophila flies with the trp mutation exhibit impaired vision due to the lack of a specific Ca2+ influx pathway in the photoreceptors. The identification of the trp gene product as a Ca2+-permeable ion channel and the search for TRP homologues in flies, worms and mammals has opened the way to the discovery of a whole superfamily of cation channels, baptized TRP channels. In contrast to voltage-gated K+, Na+, or Ca2+ channels, with whom they share their transmembrane architecture, TRP channels are not activated by voltage but by a variety of signals including intra- and extracellular ligands, Ca2+-store depletion and mechanical or thermal stress. Due to the promiscuity of these gating mechanisms, TRP channels are privileged candidates as primary sensing molecules for the recognition and integration of physical and chemical signals from the environment. In this review we discuss recent evidence that implicates members of the TRP superfamily in sensory signal transduction. PMID- 12647030 TI - Osmotic stress alters the intracellular distribution of non-erythroidal spectrin (fodrin) in bovine aortic endothelial cells. AB - Cell swelling is known to result in unfolding of membrane invaginations and restructuring of F-actin. The effect of cell swelling on the intracellular distributions of other cytoskeletal proteins that constitute the submembrane cortical cytoskeleton is virtually unknown. This study focuses on the effects of cell swelling on non-erythroidal spectrin (fodrin, also known as spectrin II), a predominant component of the membrane cytoskeleton. The intracellular distribution of spectrin in vascular endothelial cells was studied by optical sectioning using a 3-D deconvolution microscopy system. Our results show that once bovine aortic endothelial cells (BAECs) reach confluency, the non erythroidal spectrin is localized in the submembrane regions of the cells. Analysis of the intensity profiles of the non-erythroidal spectrin under isotonic and hypotonic conditions show that: (a) the width of the submembrane spectrin staining increases gradually with time within the first 5 minutes after the osmotic shock; (b) significant recovery is observed after 10 minutes even if the cells are maintained in hypotonic medium, and (c) spectrin distribution is altered by disrupting F-actin with latrunculin A but not by stabilizing F-actin with jasplakinolide. We suggest that cell swelling results in partial translocation of the submembrane spectrin to the cytosol and that it may play a major role in initiation of swelling-induced cellular events. PMID- 12647031 TI - Reactive oxygen species regulate swelling-induced taurine efflux in NIH3T3 mouse fibroblasts. AB - NIH3T3 mouse fibroblasts generate reactive oxygen species (ROS) and release taurine following exposure to hypotonic medium and to isotonic medium containing the lipase activator melittin. The swelling-induced taurine release is potentiated by H2O2, the calmodulin antagonist W7, and ATP, but inhibited by the antioxidant butulated hydroxytoluene (BHT), the NAD(P)H oxidase inhibitor diphenylene iodonium (DI), and the iPLA2 inhibitor bromoenol lactone (BEL). The swelling-induced ROS production is also inhibited by BHT and BEL. H2O2 does not affect the volume set point for activation of the volume-sensitive taurine efflux. The 5-lipoxygenase (5-LO) inhibitor ETH 615-139 impairs the swelling induced taurine efflux in the absence as well as in the presence of H2O2. The melittin-induced taurine release is, in analogy with the swelling-induced taurine release, potentiated by H2O2 and inhibited by BHT, DI, BEL, ETH 615-139 and anion channel blockers. Thus, swelling- and melittin-induced cell signalling and taurine release involve joint elements. The swelling-induced taurine efflux is potentiated by the protein tyrosine phosphatase inhibitor vanadate, and the potentiating effect of H2O2 and vanadate is impaired in the presence of protein tyrosine kinase inhibitor genistein. It is suggested that (i) iPLA2 and 5-LO activity is required for the swelling-induced activation of taurine efflux from NIH3T3 cells, (ii) ROS are produced subsequent to the PLA2 activation by the NAD(P)H oxidase complex, and (iii) ROS inhibit a protein tyrosine phosphatase (PTP1B) causing a potentiation of the swelling-induced taurine release. PMID- 12647032 TI - Fusion, leakage and surface hydrophobicity of vesicles containing phosphoinositides: influence of steric and electrostatic effects. AB - Calcium and lanthanum ion-induced fusion of lipid vesicles containing phosphatidylinositol (PI), phosphatidylinositol-4-monophosphate (PIP), phosphatidylinositol-4,5-bisphosphate (PIP2) or phosphatidylinositol-3,4,5 trisphosphate (PIP3) and its associated membrane properties, e.g., surface dielectric constant and vesicle leakage, were studied by fluorescence methods. The presence of poly-phosphorylated phosphoinositides (PPI) in lipid vesicles enhanced fusion, depending on the PPI phosphorylation level and the PPI concentration, as determined by the lipid mixing assay. This correlation held even at physiologically relevant small concentrations of PPI in vesicle membranes. However, the presence of nonphosphorylated PI inhibited fusion due to the steric effect of the inositol ring. The cation threshold concentration for the lipid mixing of vesicles made of mixtures of phosphatidylserine (PS) with PI increased with increasing PI contents. For all vesicle systems studied, a decrease in vesicle surface dielectric constant and an increase in vesicle leakage accompanied fusion. The presence of the nonphosphorylated inositol ring in PI did not interfere with the changes in the surface dielectric constant caused by fusogenic cations. Therefore, we deduce that the reduction of the surface dielectric constant is a necessary condition for membrane fusion to occur but it does not correlate with membrane fusion when interacting membranes are blocked for close approach as by the nonphosphorylated inositol ring. PMID- 12647033 TI - Gating models of the anomalous mole-fraction effect of single-channel current in Chara. AB - The dependence of single-channel current on the Tl+/K+ mole fraction exhibiting a minimum at [Tl+]/[K+] of about 1:15 is proportional to open probability in bursts. Five models are suggested to explain modulation of gating by the Tl+/K+ ratio. Three models start from a channel with 4 identical subunits, each with an allosteric binding site for K+ or Tl+. In the first model, ion binding is directly observable as a transition from one Markov state to another. This model can explain the dependence of the apparent single-channel current on Tl+ concentrations. However, the predicted linear dependence on ion concentrations of the apparent rate constants was not observed in measurements in 25 or 250 mM KNO3 and 250 mM Tl NO3. The second model can overcome this problem by introducing saturation kinetics for ion binding. In the third model, gating is caused by inherent vibrations of the protein, and the rate constants of the related transitions depend on the occupation of the allosteric sites. The fourth model is based on the foot-in-the-door approach with the essential feature that two K+ ions in the selectivity filter are necessary to keep the pore radius suitable for K+ ions. The fifth model is also a foot-in-the-door model, but non-Markovian because, similar to model 3, it is assumed that the conformation of the protein (and thus the rate constants of the Markov model of the time series) depends on the force exerted by the temporal average over the states of a Markov model of ion occupation. These ions may reside in the pore itself or outside. PMID- 12647034 TI - External copper inhibits the activity of the large-conductance calcium- and voltage-sensitive potassium channel from skeletal muscle. AB - We have characterized the effect of external copper on the gating properties of the large-conductance calcium- and voltage-sensitive potassium channel from skeletal muscle, incorporated into artificial bilayers. The effect of Cu2+ was evaluated as changes in the gating kinetic properties of the channel after the addition of this ion. We found that, from concentrations of 20 microM and up, copper induced a concentration- and time-dependent decrease in channel open probability. The inhibition of channel activity by Cu2+ could not be reversed by washing or by addition of the copper chelator, bathocuproinedisulfonic acid. However, channel activity was appreciably restored by the sulfhydryl reducing agent dithiothreitol. The effect of copper was specific since other transition metal divalent cations such as Ni2+, Zn2+ or Cd2+ did not affect BK(Ca) channel activity in the same concentration range. These results suggest that external Cu2+-induced inhibition of channel activity was due to direct or indirect oxidation of key amino-acid sulfhydryl groups that might have a role in channel gating. PMID- 12647036 TI - Isolation of mutants of Vibrio anguillarum defective in haeme utilisation and cloning of huvA, a gene coding for an outer membrane protein involved in the use of haeme as iron source. AB - The isolation of Vibrio anguillarum mutants lacking the ability to use haemin and haemoglobin as the only iron sources, as well as the identification of a gene involved in haeme utilisation are described. One of the isolated mutants defective in haeme utilisation lacked an iron-regulated outer membrane protein of 79-kDa. Although growth on haeme as iron source was completely abolished, the haemin and haemoglobin binding activities remained intact in the mutant, suggesting that the absent protein is not the only one involved in haeme binding. The wild-type phenotype in this mutant was restored by transformation with a cosmid clone (pML1) containing a 21-kb DNA fragment isolated from a gene library derived from the parental strain of V. anguillarum. Sequence analysis of pML1 subclones led to the finding of an ORF, huvA, that codes for a 79-kDa protein (HuvA) and whose sequence shows high identity with haeme receptors from Vibrio choleare (HutA) and Vibrio vulnificus (HupA). The sequence of huvA from the V. anguillarum haeme-utilisation mutant revealed a single mutation, leading to the synthesis of a truncated HuvA protein of 70 kDa. The parental strain and the cosmid-complemented mutant showed a higher degree of virulence for fish than the mutant strain in experimental infections in which fish were previously overloaded with haemin. This finding suggests that haeme uptake plays an important role in V. anguillarum multiplication in fish tissues when free haeme is available. PMID- 12647037 TI - Age of red blood cell transfusions in critically ill patients: comparison of two opposite transfusion policies. PMID- 12647038 TI - Lipopolysaccharide preconditioning attenuates metabolic alteration induced by endotoxin shock: tissue-specific monitoring by microdialysis. PMID- 12647039 TI - [Interdisciplinary treatment of diabetic foot syndrome]. AB - The amputation rate in patients with diabetic foot syndrome (DFS) in Germany is still as high as 28,000 per year. Ischemia and osteomyelitis often complicate the DFS. Impaired wound healing frequently requires further surgery with a higher amputation level. The results of treating patients with DFS in our specialized foot care center were evaluated in order to assess our interdisciplinary strategy. Advanced diabetic foot wounds in patients with ischemia and osteomyelitis first require diagnostics concerning polyneuropathy, osteomyelitis, and blood supply. If peripheral arterial vessel disease is present, surgical revascularization by distal bypass grafting is the first and crucially important element of the interdisciplinary approach. Minor amputation or elective resection of the infected bone improves wound healing. Post-interventional care for wounds with secondary healing and prevention of new ulcers are provided in a foot care clinic specialized in diabetes. The clinical and radiological results of 77 patients who underwent this treatment algorithm including bypass surgery and bone resection within 1 year were collected using a standardized questionnaire. Those results were subjected to a historical comparison. Only three patients needed further intervention because of persisting ulcers and osteomyelitis. The frequency of major amputations in all patients with DFS and ischemia combined with osteomyelitis was low (10.3%). This interdisciplinary concept of treatment guarantees a high healing rate in patients even with osteomyelitis and ischemia and allows the reduction of the rate of major amputations. The data obtained allow a fact-based design for future studies. PMID- 12647040 TI - [Effect of body weight on plantar peak pressure in diabetic patients]. AB - The goal was a better understanding of the correlation between body mass and plantar peak pressure in patients with diabetic neuropathy. A further aim was to obtain insight into the practicability of simulated weight increase and weight release and to delineate more clearly the indications for reducing body weight in order to lower the risk of ulcerations. Simulated change of body weight: (1) +20 kg by a waistcoat and (2)-20 kg by a movable overhead suspension. While normal forces and thus mean normal pressures depend directly on body mass, this does not necessarily apply to peak normal pressures. As shown in a transversal study by Cavanagh with pressure measurements in the bare foot, there is only a poor correlation between body mass and peak pressure. Therefore, peak pressure in heavy persons cannot be expected to be higher than in lighter weight persons. However, from these results it cannot be concluded that peak pressure is likely to remain unchanged irrespective of a change in body weight in a specific subject. Ten subjects were investigated: five controls and five patients suffering from diabetes without neuropathy or preceding ulcerations. All subjects wore the same kind of ready made shoes with ready made standard fitting insoles of cork. Each subject was measured in three modes of weight simulation: normal weight,20 kg weight increase (waistcoat with weight pieces), and 20 kg weight release. Weight release was effected by a modified rescue harness attached to an overhead suspension rail with 6m free walking distance. Individual alignment of the waistcoat and the suspension was checked by a force platform. In-shoe pressure measurement was done with the PEDAR in-shoe system (by Novel, Munich,Germany). For data analysis with the PEDAR standard software only peak pressures were considered. The foot was divided into six regions, particularly metatarsal region and heel. No significant difference between diabetics and controls was found. In the regions at highest risk (metatarsals and heel),peak pressure increased and decreased with weight. In the combined group (n=10), a simulated weight loss of 20 kg decreased metatarsal peak pressure by 5.4+/-1.9 N/cm(2), a 20 kg weight gain increased it by 7.4+/-5.1 N/cm(2). Therefore, without a significant deviation from linearity, peak pressure was found to be a linear function of weight. The results of this study show that weight increase or weight loss in the individual patient has an effect on the plantar peak pressure. The effect is significant in the metatarsal and heel regions. The linearity allows for a simple method of predicting the effect of weight loss by inverting the effect of simulated weight gain. PMID- 12647041 TI - [Effect of weight load and carrying conditions on plantar peak]. AB - Knowledge is limited regarding the effects exerted by carrying a load on the distribution of plantar peak pressures. It is unknown whether a special technique might exist which keeps peak pressures low when carrying a load. This question is important to diabetic neuropathic patients at risk for tissue damage and pressure ulcerations and therefore with a need for minimized plantar peak pressures. The study included 19 healthy volunteers (14 M, 5 F, mean age: 34.2+/-15.2 years). They walked three times each along a 7-m walkway with an EMED pressure measurement platform under five different carrying conditions. The conditions were: carrying no load (reference), carrying a load of 20 kg in a backpack, carrying a load of 20 kg in a waistcoat, and carrying 20 kg in two bags. A fifth condition, carrying a waistcoat with a load of 10 kg only,was used to check the linearity of peak pressure and weight. Velocity and step length were measured to check the gait parameters. Peak pressures were determined in six regions: large toe, small toes,metatarsals, lateral and medial midfoot, and heel. In agreement with the literature, the reference measurements showed peak pressure to be widely independent of body weight. On the other hand, a significant increase of peak pressure was observed in a single person when a carried load was added. In the regions of the large toe,metatarsals, and heel the peak pressure increased linearly with the carried weight and amounted to 0.54, 0.76, and 0.38 N/cm(2) per kg additional load, respectively. No significant difference between the various techniques of carrying was detected. The plantar peak pressure increases with weight load,however, this is independent of the way the load is carried. No specific recommendation on the technique of carrying a load can be given in order to take care of diabetic feet. PMID- 12647042 TI - [The MRSA patient in technical orthopaedics and rehabilitation]. AB - Colonisation and infection of humans by methicillin-resistant Staphylococcus aureus (MRSA) was examined retrospectively at the Clinic of Technical Orthopedics and Rehabilitation of the University Clinic at Munster (Germany). The cohort consisted of 28 patients who over the period were microbiologically shown to harbour MRSA from January 1997 to June 2000. Out of these, only 16 patients were colonised and only 12 patients developed MRSA infection. The inpatient stay was longer for MRSA subjects than for non-MRSA subjects. All patients had chronic wound healing disorders of the lower extremities which were due to peripheral neuropathies, diabetes mellitus and/or obstructive disease of the arteries. All patients were at risk for colonisation with MRSA. PMID- 12647043 TI - [The MRSA patient in technical orthopaedics and rehabilitation. Part 2: Hygiene management]. AB - One of the multiresistant pathogens is methicillin-resistant Staphylococcus aureus (MRSA),which was increasingly found in the 1980's. In the USA, the ratio of MRSA rose from 2% in 1975 to 29% in 1991. A similarly steep rise was seen in Germany, with the Paul Ehrlich Society for Chemotherapy reporting a rise from 1.7% in 1990 to 15.7% in 1998. In order to control MRSA, the Clinic and Polyclinic of Technical Orthopaedics and Rehabilitation at the University Clinic Munster, Germany,has set up a comprehensive hygiene management programme for early detection and treatment of nosocomial infections. In addition to addressing the usual problems of MRSA carriers (e.g., higher mortality, longer hospital stays, stigmatisation), these preventive measures are also aimed at combating MRSA-caused cost increases. PMID- 12647044 TI - [Brunner's technique for open wedge resection]. AB - The interaction of different risk factors exposes the feet of diabetics to a very high risk of infection. Therapeutic measures that are often implemented too late or are inappropriate result in major amputation. A retrospective analysis was performed to assess whether surgical reconstruction with an open wedge resection could also be achieved in cases of serious infections. An open wedge resection was performed between 1996 and 2001 on ten patients who had been suffering from diabetes mellitus for an average of 14 years. After V-shaped excision of necrotic tissue and metatarsal osteotomy close to the base, the wound was treated with wet compresses. The patients were mobilized with interim shoes or orthotics. None of the patients with an average age of 45 years experienced a recurrent ulcer or infection during the average follow-up period of 19 months. Eight patients were fitted with made-to-measure orthopedic shoes while modified standard shoes sufficed for two patients. Brunner's technique for open wedge resection is adequately radical to repair deep forefoot defects and simultaneously appropriate to avoid major amputation. The afflicted patients thus retain the highest measure of mobility and quality of life. PMID- 12647045 TI - [Link-Witzel operation for diabetics]. AB - The inner Chopart's amputation for tuberculosis of the tarsal as performed by Link and Witzel at the end of the nineteenth century laid the groundwork for a foot-saving surgical procedure in cases of neuropathic osteoarthropathy with concomitant infection. The question was posed whether such an inner amputation can represent an alternative to the classic hindfoot amputation. Six patients with neuropathic osteoarthropathy, five of whom had diabetes mellitus,who had been operated between 1989 and 2002 were analyzed retrospectively. In all of the cases, surgery had been indicated by chronic osteomyelitis of the tarsal with concomitant, extensive, mainly plantar ulcerations. During the average follow-up period of 55 weeks, no recurrence of an ulcer or infection occurred and therefore no further proximal amputation was necessary. In addition to being provided with orthotics, the remaining five patients could be fitted with made-to-measure orthopedic shoes. In selected cases, the inner Chopart's amputation can represent a solution even in such problematic cases that cannot be solved by a typical hindfoot amputation. The surgical procedure according to Link and Witzel should thus not only be considered as an alternative to Chopart's amputation, but also as a further method to spare diabetics major amputations. PMID- 12647046 TI - [Sequelae of diabetes mellitus in the vertebral column and spinal cord]. AB - Diffuse idiopathic skeletal hyperostosis (DISH) is a chronic disease of the spine characterized by a significant association with metabolic alterations such as diabetes mellitus. Diabetes decreases the immunocompetence and increases the susceptibility to infections such as spondylitis, spondylodiscitis, and epidural abscess with spinal cord lesions in a high percentage of the cases. Microangiopathic changes of the intrinsic vasculature of the spinal cord are rare and mild. In a retrospective study of 784 acute spinal cord lesions, suspected diabetes could be found in only 13 patients. Neurophysiological and animal experiments as well as pathological studies have provided proof for the presence of diabetic myelopathy. Diabetic neuroarthropathy of the spine is rare, but important in the differential diagnosis of spinal disorders. PMID- 12647047 TI - [Influence of psychic comorbidity on the treatment process of patients with diabetic foot ulcer]. AB - Until now there are only few studies with small samples dealing with psychosocial aspects of patients with diabetic foot ulcer. The main interest concerns the quality of life of these patients. By reviewing the literature and our own clinical experience, the influence of psychic comorbidity on the treatment process will be demonstrated. Mobility and hospitalization are also related to problems of coping with diabetic foot ulcer and psychosocial problems (e.g., job, partnership, and social support). Therefore, disease management programs for patients with diabetes mellitus types I and II should include psychosomatic psychotherapeutic diagnostics. The necessity for interdisciplinary work between orthopedics and psychosomatics is discussed. PMID- 12647048 TI - [Fat embolism during total hip arthroplasty. Cementless versus cemented--a quantitative in vivo comparison in an animal model]. AB - Thrombembolic complications,which include the fat embolism syndrome, are well known consequences of cementless and cemented femoral total hip replacement. Thrombembolic phenomena have been demonstrated in clinical and experimental situations with both these fixation techniques, but so far no exact quantification of the intravasated fat emboli has been performed. In a standardized animal model in 15 Merino sheep we investigated the intravasation of fat into the bloodstream during simultaneous bilateral prosthetic implantation (cemented versus cementless). After identical preparation of the intramedullary canal on both sides, a cement restrictor was additionally inserted on the cemented side and the canal was cleaned by 250 ml jet lavage. Catheters in the external iliac veins made it possible to collect the drained blood in two phases, after preparation of the intramedullary canal and during insertion of the prosthesis, and the fat content of these blood samples was measured. The amount of fat that passed into the venous draining system of the femur induced by cemented implantation (2.2749 g; S=+/-1.0079) was twice the amount seen with cementless implantation (1.1586 g; S=+/-0.4555) ( P=0.0002). An obvious effect of the canal preparation was recognizable with the cemented implantation, 8 of the 13 animals evaluated showing a peak in the fat intravasation caused by application of the cement restrictor. Our results emphasize the importance of a thorough preparation of the intramedullary canal, particularly when cemented fixation is performed. The jet lavage,which should be considered mandatory standard in cemented total hip arthroplasty, should be implemented before the insertion of the cement restrictor in order to further reduce the risk of fat embolism. PMID- 12647049 TI - [Pediatric flexible flatfoot]. PMID- 12647050 TI - Construction of a linkage map and QTL analysis of horticultural traits for watermelon [Citrullus lanatus (THUNB.) MATSUM & NAKAI] using RAPD, RFLP and ISSR markers. AB - We have been constructing linkage maps for watermelon ( Citrullus lanatus) on the basis of random amplified polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP), inter-simple sequence repeats (ISSRs) and isozymes using an F(2) population derived from a crossing between a cultivated inbred line (H-7; C. lanatus) and an African wild form (SA-1; C. lanatus). A total of 120 F(2) plants was used for construction of a linkage map using 477 RAPDs, 53 RFLPs, 23 ISSRs and one isozyme markers. Linkage analysis revealed that 554 loci could be mapped to 11 linkage groups that extended for 2,384 centimorgans (cM). While a BC(1) population [(H-7 x SA-1) x H-7] consisting of 60 individuals was grown and scored for quantitative traits. Another linkage map with a total length of 1,729 cM was constructed in the BC(1) using genetic markers found to segregate in the F(2) population. A QTL analysis was applied by means of interval mapping for locating such agronomic traits as hardness of rind, Brix of flesh juice, flesh color (red and yellow) and rind color. The relative order of markers in the BC(1) map was essentially the same as that on the linkage map in the F(2). A total of five QTLs for four agronomic traits was detected. The QTL for hardness of rind was mapped on group 4. The linkage group 8 contained the QTL for sugar content of the flesh as expressed in Brix of the juice. The QTL for red flesh color was detected on groups 2 and 8. The QTL for rind color mapped on the group 3. The present map and QTL analysis may provide a useful tool for breeders by introducing valuable wild watermelon genes to cultivars. PMID- 12647052 TI - Identification of five new blast resistance genes in the highly blast-resistant rice variety IR64 using a QTL mapping strategy. AB - Rice progenies used for the construction of genetic maps permit exhaustive identification and characterization of resistance genes present in their parental cultivars. We inoculated a rice progeny derived from the cross IR64 x Azucena with different Magnaporthe grisea isolates that showed differential responses on the parental cultivars. By QTL mapping, nine unlinked loci conferring resistance to each isolate were identified and named Pi-24( t) to Pi-32( t). They could correspond to nine specific resistance genes. Five of these resistance loci (RLs) were mapped at chromosomal locations where no resistance gene was previously reported, defining new resistance genes. Using degenerate primers of the NBS (nucleotide binding site) motif found in many resistance genes, two resistance gene analogues (RGAs) IR86 and IR14 were identified and mapped closely to two blast RLs (resistance identified in this study, i.e. Pi-29(t) and Pi-30(t) respectively). These two RLs may correspond to the Pi-11 and Pi-a blast resistance genes previously identified. Moreover, the ir86 and ir14 genes have been identified "in silico" on the indica rice cultivar 93-11, recently sequenced by Chinese researchers. Both genes encodes NBS-LRR-like proteins that are characteristics of plant-disease resistance genes. PMID- 12647051 TI - Isolation and characterization of a full-length resistance gene homolog from soybean. AB - Using mixed resistance gene analogs as probes, a putative resistance gene (KR1) was isolated from soybean and characterized further. The KR1 protein consists of a Toll/interleukin receptor (TIR) domain, a nucleotide binding site (NBS) domain, an imperfect leucine-rich repeat (LRR) domain and two C-terminal transmembrane segments. Due to these features, KR1 represents a distinct member in the TIR-NBS LRR class of resistance genes. Southern-blot analysis indicated that there were several KR1-related sequences within the soybean genome, and two polymorphic loci were mapped onto linkage group L. KR1 was induced by SA treatment and soybean mosaic virus (SMV) infection in the resistant line (Kefeng 1). An orthologue (NR1) and a homologue (NR2) of the KR1 gene were also identified in the SMV susceptible-line Nannong1138-2. Sequencing analysis revealed that NR2 was highly homologous to KR1 and NR1, but had a 21-bp deletion. Moreover, the NR1, NR2 transcription and the ratio of NR1/ NR2 was up-regulated by viral infection in Nannong1138-2. These results indicated the complexity of the regulatory mechanism in the plant responses to SMV infection. PMID- 12647053 TI - Multiple origins of allopolyploid Aegilops triuncialis. AB - Polyploidization is a key component of plant evolution. The number of independent origins of polyploid species traditionally has been underestimated. The objective of this study was to ascertain the number of origins of a tetraploid Aegilops species. We screened 84 primer sets to identify genome-specific primer sets for the tetraploid wheat relative [Aegilops triuncialis (UUCC genome)] and its diploid progenitors [Ae. umbellulata (UU genome) and Ae. caudata (CC genome)]. Primer sets G12 and G43 were U genome-specific and D21 was a C genome-specific primer. DNA sequence comparison of the G43 locus was used to estimate the number of polyploidization events in the formation of Ae. triuncialis. Parsimony analysis of G43 data revealed at least two independent formations of Ae. triuncialis. In the chloroplast hotspot region, located between genes rbcL and petA, sequence analysis suggested that at least three polyploidization origins might have occurred independently. Ae. triuncialis appears to be a tetraploid derived from multiple origins with minimal genome change after its formation. PMID- 12647054 TI - Sequence variability of a dehydrin gene within Helianthus annuus. AB - Dehydrins are proteins produced during the late stages of plant embryo development and following any environmental stimulus involving dehydration. In order to investigate the variability of a dehydrin-encoding gene (Dhn1) in cultivated and wild sunflower (Helianthus annuus) genotypes, near-complete alleles were isolated by the polymerase chain reaction and sequenced. All of the isolated sequences were found to contain the typical dehydrin domains, and interrupted by an intron. The number of nucleotide substitutions and indels per site was calculated. With respect to the overall sequence, variation in both the coding and noncoding [intron and 3'-UTR (untranslated region)] sequences was much larger among wild accessions than among cultivars. No variation was observed in 3'-UTRs from cultivated sunflowers. Different coding regions showed a different numbers of synonymous and nonsynonymous substitutions. The Y and K domains were the most conserved in both wild and cultivated genotypes. Sequence analysis of the deduced dehydrin proteins showed that nucleotide substitutions in wild accessions should also determine large biochemical differences at the protein level. All of the isolated alleles were however functional, at least at the transcription level. To our knowledge these are the first data on intraspecific genetic variability of such a stress response gene. The low variability of dehydrin genes from cultivated sunflower is discussed in relation to the origin of sunflower cultivars. The possibility of rescuing general genetic variability through crosses to wild accessions of H. annuus rather than using wild Helianthus species is also discussed. PMID- 12647055 TI - A set of simple-sequence repeat (SSR) markers covering the Prunus genome. AB - A set of 109 microsatellite primer pairs recently developed for peach and cherry have been studied in the almond x peach F(2) progeny previously used to construct a saturated Prunus map containing mainly restriction fragment length polymorphism markers. All but one gave amplification products, and 87 (80%) segregated in the progeny and detected 96 loci. The resulting Prunus map contains a total of 342 markers covering a total distance of 522 cM. The approximate position of nine additional simple sequence repeats (SSRs) was established by comparison with other almond and peach maps. SSRs were placed in all the eight linkage groups of this map, and their distribution was relatively even, providing a genome-wide coverage with an average density of 5.4 cM/SSR. Twenty-four single-locus SSRs, highly polymorphic in peach, and each falling within 24 evenly spaced approximately 25-cM regions covering the whole Prunus genome, are proposed as a 'genotyping set' useful as a reference for fingerprinting, pedigree and genetic analysis of this species. PMID- 12647056 TI - Comparison between Poncirus and Citrus genetic linkage maps. AB - Five genetic linkage maps were constructed for the parents of three progenies: Citrus aurantium (A) x Poncirus trifoliata var. Flying Dragon (Pa), C. volkameriana (V) x P. trifoliata var. Rubidoux (Pv) and a self-pollination of P. trifoliata var. Flying Dragon (Pp). The number of polymorphic markers assayed ranged from 48 for Pa to 120 for A according to the heterozygosity of each parental. As our focus was on genome comparison, most of the markers were newly generated simple sequence repeats. Inter-retrotransposon amplified polymorphisms based on four retrotransposon sequences isolated from Citrus spp were also used to saturate the maps. These polymorphisms were much more frequent in A (53) than in Pa (15) and randomly distributed throughout both genomes. Since comparative genomics and quantitative trait locus analysis applicability depends on the reliability of marker ordering, the causes of variation in marker order were investigated. Around 25% of the markers showed gametal segregation distortions. Segregation distortions were also observed at the zygotic level towards a reduction in the observed frequency of homozygotes from that expected in linkage groups 5 and 7. The presence of balanced lethal factors or gametal incompatibility genes in those genomic regions would explain a zygotic advantage of heterozygotes at these specific regions. Four differences in genomic organization were observed; three are putative translocations and affect homeologous linkage groups 3, 7 and 11, where highly distorted markers are found. Other causes of variation in marker order are also discussed: the introduction of new markers in the map, lowering the LOD score and the mapping software. These results represent the first comparative mapping analysis among Citrus and Poncirus species. PMID- 12647057 TI - Pollen morphology and in vitro germination characteristics of nodulating and nonnodulating soybean (Glycine max L.) genotypes. AB - Artificial hybridization in highly self-pollinated crop species such as soybean (Glycine max L.) is important for both generating genetic variability and segregation for selection. In higher plants, pollen is an agent for transmission of genetic information over generations. The objective of this study was to measure and compare both morphological (length, width) and in vitro germination (germination percentage tube length) characteristics of pollen from the nodulating soybean cultivar, Bragg, and a nonnodulating Bragg mutant line, Nod 139, obtained following ethyl methanesulphonate treatment. Highly significant (P = 0.007) differences in pollen length were observed between these two genotypes. Similarly, in vitro germination percent (G%) indicated highly significant (P = 0.0001) differences between these genotypes, suggesting that the nodulation trait produces variation in in vitro germination capacity of the pollen. It appears the nonnodulation trait in soybean alters pollen grain length and G%. PMID- 12647058 TI - Determining the sample size for co-dominant molecular marker-assisted linkage detection for a monogenic qualitative trait by controlling the type-I and type-II errors in a segregating F2 population. AB - Tests for linkage are usually performed using the lod score method. A critical question in linkage analyses is the choice of sample size. The appropriate sample size depends on the desired type-I error and power of the test. This paper investigates the exact type-I error and power of the lod score method in a segregating F(2) population with co-dominant markers and a qualitative monogenic dominant-recessive trait. For illustration, a disease-resistance trait is considered, where the susceptible allele is recessive. A procedure is suggested for finding the appropriate sample size. It is shown that recessive plants have about twice the information content of dominant plants, so the former should be preferred for linkage detection. In some cases the exact alpha-values for a given nominal alpha may be rather small due to the discrete nature of the sampling distribution in small samples. We show that a gain in power is possible by using exact methods. PMID- 12647059 TI - Genetic and morphological characterization of the barley uniculm2 (cul2) mutant. AB - Axillary meristem growth and development help define plant architecture in barley (Hordeum vulgare L). Plants carrying the recessive uniculm2 (cul2) mutation initiate vegetative axillary meristem development but fail to develop tillers. In addition, inflorescence axillary meristems develop into spikelets, but the spikelets at the distal end of the inflorescence have an altered phyllotaxy and are sometimes absent. Double mutant combinations of cul2 and nine other recessive mutations that exhibit low to high tiller number phenotypes resulted in a uniculm vegetative phenotype. One exception was the occasional multiple shoots produced in combination with granum-a; a high tillering mutant that occasionally produces two shoot apical meristems. These results show that the CUL2 gene product plays a role in the development of axillary meristems into tillers but does not regulate the development of vegetative apical meristems. Moreover, novel double-mutant inflorescence phenotypes were observed with cul2 in combination with the other mutants. These data show that the wild-type CUL2 gene product is involved in controlling proper inflorescence development and that it functions in combination with some of the other genes that affect branching. Our genetic analysis indicates that there are genetically separate but not distinct regulatory controls on vegetative and inflorescence axillary development. Finally, to facilitate future positionally cloning of cul2, we positioned cul2 on chromosome 6(6H) of the barley RFLP map. PMID- 12647060 TI - Microsatellite mapping of the powdery mildew resistance gene Pm5e in common wheat (Triticum aestivum L.). AB - Powdery mildew, caused by Erysiphe graminis DM f. sp. tritici (Em. Marchal), is one of the most important diseases of common wheat world-wide. Chinese wheat variety 'Fuzhuang 30' carries the powdery mildew resistance gene Pm5e and has proven to be a valuable resistance source of powdery mildew for wheat breeding. Microsatellite markers were employed to identify the gene Pm5e in a F(2) progeny from the cross 'Nongda 15' (susceptible) x 'Fuzhuang 30' (resistant). The gene Pm5e was mapped in the distal region of chromosome 7BL. Seven microsatellite markers were found to be linked to the gene Pm5e, of which two codominant markers Xgwm783 and Xgwm1267 were relatively close to Pm5e with a linkage distance of 11.0 cM and 6.6 cM, respectively. It is possible to use the 136-bp allele of Xgwm1267 in 'Fuzhuang 30' for marker-assisted selection during the wheat resistance breeding process for facilitation of gene pyramiding. The mapping information in the present study provides a starting point for fine mapping of the Pm5 locus and map-based cloning to clarify the molecular structure and function of the different alleles at the Pm5 locus. A microsatellite linkage map of chromosome 7B was constructed with 20 microsatellite loci, nine on the short arm and 11 on the long arm. This information will be very useful for further mapping of agronomically important genes of interest on chromosome 7B. PMID- 12647061 TI - Genetic mapping and analysis of quantitative trait loci affecting fiber and lignin content in maize. AB - Plant cell walls of forage provide a major source of energy for ruminant animals. Digestion of cell walls is limited by the presence of lignin, therefore the improving the digestibility of forages by reducing lignin content is a major goal in forage crop breeding programs. A recombinant inbred line maize population was used to map quantitative trait loci (QTL) for neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL) of leaf-sheath and stalk tissues. All traits were positively genetically correlated. The larger genetic correlations were between NDF and ADF in sheaths (r = 0.84), NDF and ADF (r = 0.96), ADF and ADL (r = 0.83), and NDF and ADL (r = 0.76) in stalks. Twelve QTL were detected for NDF and 11 QTL for ADF in leaf-sheaths. Eight QTL detected for both traits were defined by the same or linked marker loci. Eight QTL were associated with leaf-sheath ADL. Eleven QTL were detected for NDF and ADF, and 12 QTL for ADL in stalks. Nine of eleven QTL detected for both NDF and ADF in stalks coincided in their genomic position. A high proportion of QTL detected for these traits had the same parental effects and genomic locations, suggesting that it is only necessary to select on one fiber component (NDF or ADF) to improve digestibility. Favorable correlated responses of unselected fiber components are expected due to coincident genomic locations of QTL and the high genetic correlation between fiber components. Several QTL detected in this study coincided in their positions with putative cellulose synthase genes from maize. PMID- 12647062 TI - High-resolution mapping of the leaf rust disease resistance gene Lr1 in wheat and characterization of BAC clones from the Lr1 locus. AB - Leaf rust is the most common disease in wheat production. There are more than 45 specific resistance genes described and used in wheat breeding to control epidemics of leaf rust, but none of them has been cloned. The leaf rust disease resistance gene 1 ( Lr1) is a good model gene for isolation by map-based cloning because it is a single, dominant gene which is located in the distal region of chromosome 5DL of wheat. As the first step towards the isolation of this gene we constructed a high-resolution genetic map in the region of the Lr1 locus by saturation mapping of two large segregating F(2) populations (Thatcher Lr1 x Thatcher, Thatcher Lr1 x Frisal). The resistance gene Lr1 was delimited in a 0.16 cM region between the RFLP markers ABC718 and PSR567 (0.12 cM from ABC718 and 0.04 cM from PSR567). A genomic BAC library of Aegilops tauschii (D genome) was screened using the RFLP markers ABC718 and PSR567. Five positive BAC clones were identified by ABC718 and four clones by PSR567. Two NBS-LRR type of resistance gene analogs, which encode proteins highly homologous to the bacterial blight disease resistance protein Xa1 of rice, were identified on BAC clones isolated with PSR567. Polymorphic BAC end probes were isolated from both ends of a 105-kb large BAC clone identified by ABC718. The end probes were mapped at the same locus as ABC718, and no recombination event was found within 105 kb around ABC718 in our analysis of more than 4,000 gametes. PMID- 12647063 TI - Cytological and breeding behavior of pentaploids derived from 3x x 4x crosses in potato. AB - Cytology and breeding behavior of Solanum commersonii - S. tuberosum hybrids derived from 3 x x 4 x crosses was examined. The chromosome number of hybrids ranged from hypo-pentaploid (2 n=5 x - 8=52), to hyper-pentaploid (2 n=5 x + 7=67), with the euploid pentaploid 2 n=5 x=60 class predominant. The high variability in chromosome number of the 3 x x 4 x hybrids was attributed to the fact that meiotic restitution during megasporogenesis of the 3 x female may have involved poles with various chromosome numbers, resulting in 2 n eggs with 24-48 chromosomes. Microsporogenesis analyses provided evidence that chromosome pairing between S. commersonii and S. tuberosum genomes occurred. In addition, chromosome distribution at anaphase I and anaphase II revealed an average chromosome number of 29.5 and 29.1 per pole, respectively. To further study the extent of transmission of extra genome chromosomes from pentaploids, 5 x x 4 x and 4 x x 5 x crosses were performed, and the chromosome number of resulting progeny was determined. Ploidy ranged from 2 n=4 x=48 to 2 n=5 x=60 following 5 x x 4 x crosses, and from 2 n=4 x + 1=49 to 2 n=5 x=60 following 4 x x 5 x crosses. These results provided indirect evidence that the pentaploid hybrids produced viable aneuploid gametes with a chromosome number ranging from 24 to 36. They also demonstrated that gametes with large numbers of extra chromosomes can be functional, resulting in sporophytes between the 4 x and 5 x ploidy level. Fertility parameters of crosses involving various (aneuploid) pentaploid genotypes were not influenced by chromosome number, suggesting a buffering effect of polyploidy on aneuploidy. The possibility of successfully using (aneuploid) pentaploid genotypes for further breeding efforts is discussed. PMID- 12647064 TI - Linkage of the A locus for the presence of anthocyanin and fs10.1, a major fruit shape QTL in pepper. AB - The purple color of the foliage, flower and immature fruit of pepper ( Capsicum spp.) is a result of the accumulation of anthocyanin pigments in these tissues. The expression of anthocyanins is controlled by the incompletely dominant gene A. We have mapped A to pepper chromosome 10 in a Capsicum annuum (5226) x Capsicum chinense (PI 159234) F(2) population to a genomic region that also controls anthocyanin expression in two other Solanaceous species, tomato and potato, suggesting that variation for tissue-specific expression of anthocyanin pigments in these plants is controlled by an orthologous gene(s). We mapped an additional locus, Fc, for the purple anther filament in an F(2) population from a cross of IL 579, a C. chinense introgression line and its recurrent parent 100/63, to the same position as A, suggesting that the two loci are allelic. The two anthocyanin loci were linked to a major quantitative trait locus, fs10.1, for fruit-shape index (ratio of fruit length to fruit width), that also segregated in the F(2) populations. This finding verified the observation of Peterson in 1959 of linkage between fruit color and fruit-shape genes in a cross between round and elongated fruited parents. The linkage relationship in pepper resembles similar linkage in potato, in which anthocyanin and tuber-shape genes were found linked to each other in a cross of round and elongated-tuber parents. It is therefore possible that the shape pattern of distinct organs such as fruit and tuber in pepper and potato is controlled by a similar gene(s). PMID- 12647065 TI - Maize introduction into Europe: the history reviewed in the light of molecular data. AB - The resolution that can be obtained from molecular genetic markers affords new prospects for understanding the dispersion of agricultural species from their primary origin centres. In order to study the introduction and the dispersion of maize in Europe, we have characterised a large and representative set of maize populations of both American and European origins for their variation at 29 restriction fragment length polymorphism loci. Polymorphism was higher for American populations than for European populations (respectively, 12.3 and 9.6 alleles per locus, on average), and only a few alleles were specific to European populations. Investigation of genetic similarity between populations from both continents made it possible to identify various types of American maize introduced into Europe at different times or in different places and which have given rise to distinctive European races. Beyond confirming the importance of Caribbean germplasm, the first maize type to be introduced into Europe, this research revealed that introductions of Northern American flint populations have played a key role in the adaptation of maize to the European climate. According to a detailed historical investigation, the introduction of these populations must have occurred shortly after the discovery of the New World. PMID- 12647066 TI - Inheritance of self-compatibility in almond: breeding strategies to assure self compatibility in the progeny. AB - To assure self-compatibility in the progenies, three different crosses were conducted for the first time in an almond breeding programme: self-pollination (266 descendants from 30 families), crosses between parents sharing an S-allele (108 descendants from five families) and crosses with homozygous self-compatible parents (62 descendants from five families). Depending on the cross, self compatibility in the progenies was determined by observing pollen tube growth (by means of fluorescence microscopy), stylar S-RNases analysis or allele-specific PCR. The results obtained fit with the accepted hypothesis of inheritance of self compatibility and the three crossing strategies used ensured 100% of self compatible descendants. These strategies increase the efficiency of the breeding programme and avoid the laborious task of evaluating this characteristic. From the breeding point of view, self-fertilisation and crosses between relatives tend to produce inbreeding. Furthermore, these methods reduce the possibilities of choosing the parental combination. The use of homozygous self-compatible parents does not have any of these disadvantages. As far as we know, this is the first time that allele-specific PCR has been used for early selection of self compatible seedlings. The advantages and disadvantages of the three methodologies used to determine self-compatibility are discussed. PMID- 12647067 TI - Development and transferability of apricot and grape EST microsatellite markers across taxa. AB - EST microsatellite markers were developed in apricot (Prunus armeniaca L.) and grape (Vitis vinifera L.). cDNA libraries from either apricot leaves or grape roots were used in an enrichment procedure for GA and CA repeats. The transferability of EST simple sequence repeat (SSR) markers from apricot and grapevine to other related and unrelated species was examined. Overall, grape primers amplified products in most of the Vitaceae accessions while the apricot primers amplified polymorphic alleles only in closely related species of the Rosaceae. In this taxonomic family, ten EST SSR loci were tested, and one single primer pair, PacB22, was amplified across species and sections in the Prunoideae and Maloideae. Sequencing of EST SSR loci in other species and genera confirmed a higher level of conservation in the microsatellite motif and flanking regions in the Vitaceae compared to the Rosaceae. Two distinct fragments of the PacB22 locus amplified across the Malus and Pyrus genera; however, while the coding region was highly conserved, the microsatellite repeat motif was no longer present. The banding pattern was explained by base substitution and insertion/deletion events in the intronic region of PacB22. This study includes the determination of the degree of polymorphism detected among species and genera in two unrelated taxonomic families and the evaluation of the information provided by the microsatellite repeats and the flanking regions. PMID- 12647068 TI - An EREBP/AP2-type protein in Triticum aestivum was a DRE-binding transcription factor induced by cold, dehydration and ABA stress. AB - We characterize one transcription factor of DRE-binding proteins (TaDREB1) that was isolated from a drought-induced cDNA library of wheat (Triticum aestivum L.). TaDREB1 contains one conserved EREBP/AP2 domain, and shows similarity with Arabidopsis thaliana DREB family members in both overall amino-acid sequences and the secondary structure arrangement within the DNA-binding motifs. In yeast one hybrid system, TaDREB1, can specially activate the genes fused with the promoter containing three tandemly repeated copies of the wild-type DRE sequence: TACCGACAT. In different wheat cultivars, the Ta DREB1 gene is induced by low temperature, salinity and drought; and the expression of Wcs120 that contains DRE motifs in its promoter is closely related to the expression of TaDREB1. These results suggest that TaDREB1 functions as a DRE-binding transcription factor in wheat. We also observed the dwarf phenotype in transgenic rice (T0) overexpressing TaDREB1. PMID- 12647069 TI - Old-growth red spruce forests as reservoirs of genetic diversity and reproductive fitness. AB - Old-growth forests are assumed to be potential reservoirs of genetic diversity for the dominant tree species, yet there is little empirical evidence for this assumption. Our aim was to characterize the relationship of stand traits, such as age, height and stem diameter, with the genetic and reproductive status of old growth and older second-growth stands of red spruce (Picea rubens Sarg.) in eastern Canada. We found strong relationships between height growth (a fitness trait) and measures of genetic diversity based on allozyme analyses in red spruce. The negative relationship between height and the proportion of rare alleles suggests that high proportions of these rare alleles may be deleterious to growth performance. Latent genetic potential, however, showed a significant and positive relationship with height. Stand age was not correlated to height, but was correlated to seedling progeny height. In late-successional species such as red spruce, age and size (e.g., height and stem diameter) relationships may be strongly influenced by local stand disturbance dynamics that determine availability of light, growing space, moisture and nutrients. In larger and older stands, age appeared to provide a good surrogate measure or indicator for genetic diversity and progeny height growth. However, in smaller and more isolated populations, these age and fitness relationships may be strongly influenced by the effects of inbreeding and genetic drift. Therefore, older populations or old growth forests may represent superior seed sources, but only if they are also of sufficient size and structure (e.g., stem density and spatial family structure) to avoid the effects of inbreeding and genetic drift. Thus, larger and older forests appear to have an important evolutionary role as reservoirs of both genetic diversity and reproductive fitness. Given the rapid environmental changes anticipated (as a result of climate change, increasing population isolation through fragmentation, or following the introduction of exotic pests and diseases) these older populations of trees may have a valuable function in maintaining the adaptive potential of tree species. PMID- 12647070 TI - Structural organization of the group-1 chromosomes of two bread wheat sister lines. AB - Eureka and Renan are two French bread wheat cultivars derived from a 4-way cross. Using molecular markers (essentially RFLPs), we studied the structure of the group-1 chromosomes of these two genotypes, their parents and a doubled-haploid (DH) population derived from their F(1). Using the DH population (102 lines), a molecular map of the three homoeologous group-1 chromosomes was produced and compared with the map established on another intervarietal cross: Courtot x Chinese Spring (Cadalen et al. 1997). The polymorphic markers were mapped on the DH population and characterized on the four grand-parents, allowing us to compare the structural organization of the group-1 chromosomes of Eureka and Renan and determine their origin. These chromosomes were very different, except for small regions (1AL proximal and 1BL distal) which were identical. PMID- 12647072 TI - [Pancreatic carcinogenesis. Clinical implications]. AB - Our understanding of the molecular pathology of pancreatic carcinoma has improved tremendously over the past few years due to the development of sophisticated molecular techniques. This knowledge has led to the postulation of a pancreatic tumor progression model. This article describes the molecular oncogenesis of pancreatic carcinoma and points out potential targets for therapeutic interventions. Pancreatic cancer is characterized by the sequential acquisition of somatic mutations in the proto-oncogene K-RAS and the tumor suppressors INK4a,TP53 and DPC4/SMAD4 and by epigenetic alterations, including the overexpression of the "epidermal growth factor" receptor/ligand system. These genetic changes cause a profound disturbance to cell cycle regulation and continuous growth. Further analysis of the underlying molecular mechanisms will offer new diagnostic and therapeutic options and hopefully improve the outcome of this grim disease in the future. PMID- 12647071 TI - Population genetic structure of Pyrenophora teres Drechs. the causal agent of net blotch in Sardinian landraces of barley (Hordeum vulgare L.). AB - Monoconidial cultures of Pyrenophora teres, the causal agent of barley net blotch, were isolated from leaves collected from six populations of the barley landrace "S'orgiu sardu" growing in five agro-ecological areas of Sardinia, Italy, and genotyped using AFLPs. The 150 isolates were from lesions of either the "net form" (P. teres f. sp. teres) or the "spot form" (P. teres f. sp. maculata) of the disease. Of 121 AFLP markers, 42%, were polymorphic. Cluster analysis resolved the isolates into two strongly divergent groups (F(ST) = 0.79), corresponding to the net (45% of the isolates) and the spot (55% of the isolates) forms (designated the NFR and SFR groups, respectively). The absence of intermediate genotypes and the low number of shared markers between the two groups indicated that hybridization between the two formae is rare or absent under the field condition of Sardinia. Five of the barley populations hosted both forms but in different proportions. The SFR populations were similar in overall polymorphism to the NFR populations. However, compared to the SFR form, the NFR occurred in all fields sampled and showed a higher population divergence (F(ST) = 0.43 versus F(ST) = 0.09 with all isolates; F(ST) = 0.37 versus F(ST) = 0.06 with clone corrected samples) probably due to a lower migration rate. AFLP fingerprints resolved 117 distinct genotypes among the 150 isolates sampled (78%), 87% in SFR and 68% in NFR isolates. Although the absolute numbers may be a function of the number of AFLP markers assayed, the relative difference suggests that clonality is more prevalent among the NFR isolates (with 11 of 46 haplotypes observed more than once), compared with SFR isolates (7 of 71 haplotypes). Both digenic and multilocus linkage disequilibrium analyses suggested that sexual reproduction occurs at significant levels within the NFR and SFR populations, and that the relative contribution of sexual and asexual reproduction varies among different environments. PMID- 12647073 TI - [Diagnostic evaluation of pancreatic carcinoma prompting treatment decisions]. AB - An early diagnosis or even sufficient screening tools may allow improved and/or more successful treatment strategies for pancreatic carcinomas. Presently,we have no useful screening tools available. The 5 year survival rates of those suffering from pancreatic cancer is around 3%, with only 15% of patients at diagnosis available for curative resection protocols. Thus, better diagnostic procedures for populations at risk are urgently required. Today,endosonography complements new CT and MRI tests. High resolution CT and contrast enhanced MRI procedures are rapidly evolving and may offer more precise diagnostic opportunities. PET tests help discriminating benign from neoplastic tumors, and new molecular markers may offer the opportunity to screen patients at risk with the hope for an early diagnosis. Nevertheless, the technical progress made in tomographic procedures is developing so rapidly that comparative studies are often outdated at publication. Therefore,we have reason to hope for improved diagnostic skills, but presently have only limited data for the new techniques currently available. PMID- 12647074 TI - [Surgical standard therapy for cancer of the pancreas]. AB - Approximately 150,000 people worldwide and 40,000 people in Europe die each year of pancreatic cancer,making it one of the five leading causes of cancer-related death and one of the most aggressive human tumors. Resection is still the only option that offers a chance of cure for pancreatic cancer patients. Recent studies have highlighted the correlation between the number of pancreatic resections per year and postoperative mortality. Thus, large centers of pancreatic surgery have mortality rates below 5%, whereas centers with lower caseloads have mortality rates exceeding 10%. Standards have been established for the surgical treatment of pancreatic cancer;however, these are often not based on evidence derived from randomized, controlled studies. Resection for pancreatic cancer is carried out if there are no metastases present and if the tumor is locally resectable; i.e., if there is no complex vessel invasion. However, an isolated infiltration of the portal vein is not considered a contraindication for surgery. At present, there are no evidenced-based data available on palliative (R2) resections,which might be a therapeutic option in centers with low morbidity/mortality. Three randomized controlled trials clearly show that the pylorus-preserving Whipple is equal to the classical Whipple in terms of oncological effectiveness, morbidity, and quality of life. Therefore, the pylorus preserving Whipple is increasingly considered the standard operation for tumors of the pancreatic head. Based on randomized trials, extended lymph node dissection cannot generally be recommended for pancreatic cancer. Further prospective, randomized, multicenter trials have to be carried out in the upcoming years to find new approaches in the therapy of pancreatic cancer and to establish evidence-based treatment strategies for this disease. PMID- 12647075 TI - [Adjuvant and additive therapy for cancer of the pancreas]. AB - Recent advances have been made in the treatment of pancreas cancer. Specialized pancreas centres have reported an increasing rate of resections with reduced postoperative mortality. On account of the highly aggressive nature of pancreas cancer, there is a great challenge in identifying effective therapy concepts for advanced stages of the cancer as well as for the development of resection associated measures. As large-scale, randomised, controlled studies are lacking, the additive therapy concepts after resection do not have a sufficiently scientific basis. The ESPAC-1 study, which included 600 patients, surpassed all previous studies on adjuvant therapy for pancreas cancer. This study has shown,for example, that the most promising adjuvant chemotherapy with 5 fluorouracil and folic acid leads to an equal if not better result than the multimodal regimen. This regimen can be superseded with the use of Gemcitabine, which will be evaluated in the ESPAC-3 study that includes 990 patients from various European countries including Germany, as well as from Canada and Australia. Participation in the large, phase-3 study therefore plays a key role in the continued development of the management of pancreas cancer. PMID- 12647076 TI - [Pancreatic cancer. The relative importance of neoadjuvant therapy]. AB - Neoadjuvant radiochemotherapy in patients suffering from pancreatic cancer is presently not well established. Neoadjuvant radiochemotherapy is recommended to be applied in 5-8 weeks. The full dose of radiotherapy is between 50 and 54 Gy with 5FU used as radiosensitator. In patients with resectable pancreatic cancer, particularly in UICC-stage II neoadjuvant radiochemotherapy, this results in an improvement in survival: the median survival is between 15 and 30 months. In about 15% of the patients with resectable pancreatic cancer (UICC I-III), neoadjuvant radiochemotherapy results in downstaging. In combination with a R0 resection,neoadjuvant radiochemotherapy effects a reduction of local recurrence. Results from controlled clinical trials are necessary to objectify the benefits of neoadjuvant radiochemotherapy. PMID- 12647077 TI - [Laparoscopic cholecystectomy. Training on a bio-simulation model with learning success documented using score-cards]. AB - The possibilities of gaining theoretical knowledge in surgery are virtually unlimited. To acquire practical skills,however, the availability and willingness of the patient are the limiting factors. With the development of training tools and structured courses, experience can be gained independently of the clinical situation. To objectify and record individual performance and improvement, and to define further potential, a scoring system is necessary. Using the example of laparoscopic cholecystectomy, a training program was combined with an evaluation system and the beginners' learning curves were recorded during the course of a training week. The present study shows that first experiences and measurable improvements can be reliably assessed within one week's structured training on a simulation model. PMID- 12647078 TI - [Endoscopic ultrasound in routine clinical practice for staging adenocarcinomas of the stomach and distal esophagus]. AB - PROBLEM: Endoscopic ultrasound (EUS) is an important diagnostic tool for determining the best therapeutic strategy (primary resection, neoadjuvant therapy or palliation only) to offer esophageal or gastric cancer patients. PATIENTS AND METHODS: In the present study (1992-2001),we evaluated the accuracy of EUS in adenocarcinomas of the distal esophagus and stomach and compared our results with pathologists findings as the gold standard. RESULTS: Of the 222 patients studied, the precise examination of 11% EUS was not completely possible due to severe tumor stenosis. The accuracy of EUS with respect to T, N+/- and TN+/- amounted to 51%, 65% and 34% in 131 patients with adenocarcinomas of the esophageal gastric junction and to 50%, 66% and 37% in 91 patients with adenocarcinomas located in the fundus, corpus or antrum of the stomach respectively. With respect to T stage, the overstaging of tumors was more common than understaging, especially in pT2b-carcinomas. The subgroup analysis of the 131 EGJ adenocarcinoma patients showed that the results obtained by EUS were slightly better in type I (distal esophageal cancer) than in type II and III cardia carcinomas (proximal gastric cancer).When comparing two observation periods (1992-1996 and 1997-2001), the accuracy of endoscopic ultrasound staging was very similar in both periods for T category (51% vs 49%) and N-category (63% vs 64%) as well as for combined TN staging (36% vs 35%) respectively. CONCLUSIONS: In clinical routine examinations of adenocarcinomas of the stomach and the distal esophagus, the accuracy of EUS is not as good as the excellent results in the past--mostly obtained under study conditions--may suggest. PMID- 12647079 TI - [Preoperative diagnostic procedures in locally advanced rectal carcinoma (> or =T3 or N+). What does endoluminal ultrasound achieve at staging and restaging (after neoadjuvant radiochemotherapy) in contrast to computed tomography?]. AB - INTRODUCTION: Neoadjuvant radiochemotherapy (neoRT/CT) in locally advanced rectal cancer requires an exact initial determination of the depth of the cancerous infiltration (T-status) and of locoregional lymph node metastasis (N-status). For staging and restaging, contrast-enhanced computed tomography (CT) is usually used. In specialised centers, the endorectal ultrasound (rES) may be preferred. METHODS: Between January 1998 and May 2001, the T- and N-status of 102 patients with adenocarcinoma of the rectum (> or =T3 or N+) was determined prospectively by rES and CT (group I: n=61 without neo-RT/CT, examined once; group II: n=41 examined before and after neoRT/CT). All diagnostic findings were compared using the (y)pTNM-classification. RESULTS: In the patients from group I, the depth of infiltration (uT) was predicted correctly by rES in 75% and by CT in 48% of cases; the carcinomas were understaged in 10% and 41% of cases and overstaged in 15% and 11%, respectively. According to the histopathological findings, the N status was determined correctly by rES and CT in 75% and 57% of cases, understaging occurred in 8% and 30% and overstaging in 17% and 13%, respectively. In cases in which both methods resulted in identical T- (uT+ctT) or N-staging (uN+ctN), the accuracy increased to 82% and 80%, respectively. In patients from group II, after neoRT/CT rES and CT allowed the exact prediction of the yuT-stage in 66% and 51%, respectively. Only 2% were understaged by rES (understaging by CT: 22%). Overstaging occurred in 32% and 27% by rES and CT, respectively. The N status determined by rES and CT was in accordance with the histopathological findings in 68% and 76%of cases, respectively. Understaging occurred in 20% and 17%,overstaging in 12% and 7%, respectively. Again identical staging results in both rES and CT increased the accuracy of the T- (yuT+yctT) or N- (yuN+yctN) classification to 90% and 83%, respectively. In group II, downsizing of the tumor by more than one T-stage was correctly assessed by rES results in 15/20 cases (75%). A complete remission of initial uT3-carcinoma was diagnosed correctly in only two of eight ypT0-cases. In contrast, CT demonstrated a remission of disease in all cases but was unable to predict the extent of tumour reduction. A remission of lymph node metastasis was accurately shown by rES in 17/19 cases (90%) and by CT in 10/12 cases (83%). CONCLUSION: The staging of pretherapeutic, locoregional T- and N-status by rES is superior to that by CT (T-status: P=0.0164, N-status: P=0.0035). At restaging, rES offers higher accuracy in the detection of residual tumour infiltration (but not significantly to CT, yT status: P=0.0833, yN-status: P=0.7962) and assessment of local remission. Therefore rES should be the method of choice in staging to avoid overtreatment in neoadjuvant settings.After neoRT/CT, the predictive efficacy of the rES for the downsizing/-staging of rectal cancer must be evaluated on greater numbers of patients receiving standardised diagnostic procedures and therapy. PMID- 12647080 TI - [Surgical pain management. A Germany-wide survey including the effect of clinical guidelines]. AB - INTRODUCTION: Over the last few years, various studies have aimed at improving the diagnosis and therapy of perioperative and posttraumatic pain. METHODS: In an anonymous postal survey, 2,393 German surgical clinics were asked to describe several aspects of their clinical pain treatment. In order to assess the influence of interdisciplinary guidelines on surgical pain therapy, the results of this survey were compared to a survey from 1997. RESULTS: The response rate was 30.8% ( n=738). Pain was measured quantitatively in 11.4% ( n=80) of hospitals, a figure which is unchanged since 1997. In pain treatment,however, some changes were discernible: while 46.6% of all hospitals in 1997 used patient controlled analgesia often or occasionally, this proportion has risen to 64.5%.Both, the national acute pain guidelines and the WHO chronic pain guidelines were well known (71.3% and 74.7%) and accepted (98.8% and 98.5%, respectively). Among those surgeons who knew the national guidelines,93.7% reported that they used them clinically. In 149 hospitals (20.2%), local guidelines have been developed either anew or from existing guidelines. CONCLUSIONS: Although clinical guidelines are widely used, pain therapy in surgical patients has improved only marginally. PMID- 12647081 TI - [Treatment of ingrown toenails. What is an "Emmert plasty"?]. AB - In Germany,"Emmert plasty" is one of the terms most frequently used in daily surgical practice. It describes the wedge excision of nail fold, nail edge, and corresponding matrix for advanced stages of ingrown toenail in most standard textbooks of surgery but without direct reference to Emmert. The method, however,was apparently first described by the French military surgeon Baudens in 1850 and was strongly opposed by Emmert himself who propagated a pure soft tissue resection. This text gives a short historical review of the various treatment methods of ingrown toenails. PMID- 12647082 TI - [Extrahepatic portal vein thrombosis following laparoscopic cholecystectomy]. AB - Postoperative portal vein thrombosis is a rare complication, which occurs most often after hepatic surgery, but has not yet been described in combination with laparoscopic cholecystectomy. We present a case where thrombotic occlusion of the portal vein was diagnosed on the 6th day following laparoscopic cholecystectomy. Treatment with high-dose anticoagulant and antibiotic therapy was successful and without further complications. PMID- 12647083 TI - [Adrenal adenomatoid tumor. A rare clinicopathological entity]. AB - Adenomatoid tumors are uncommon, benign tumors of the genital tract which have also been reported to occur extragenitally. Case reports on adenomatoid tumors of the adrenal gland exist. Most of these are incidentally discovered at autopsy or after the resection of incidentalomas. We report on the case of a young man with epigastic pain and with the finding of a 4 cm heterogeneous right adrenal mass on abdominal CT scan. After endocrine activity had been ruled out, an inactive, benign adrenal tumor was suspected and laparoscopic right adrenalectomy performed. The specimen was found to be an adenomatoid tumor. We discuss the differential diagnosis and the possible embryological origin of these tumors. The feature of 'local invasive ability' does not imply malignancy. All cases discovered surgically and at autopsy have been benign. Local resection seems to be the appropriate therapy. PMID- 12647084 TI - [Postoperative pain therapy]. PMID- 12647086 TI - [Cost analysis and outcome prediction with the Therapeutic Intervention Scoring System (TISS and TISS-28]. AB - BACKGROUND AND PURPOSE: The "Therapeutic Intervention Scoring System" (TISS) and the simplified version TISS-28 obtain the therapeutic workload in the critically ill and may be used for outcome prediction. The feasibility and applicability regarding cost analysis and outcome prediction of TISS and TISS-28 have been assessed in 303 consecutive medical patients staying longer than 24 h in the intensive care unit (ICU). PATIENTS AND METHODS: The mean age of the enrolled patients was 62 +/- 12 years, 216 (71%) patients were male, length of ICU stay 3.7 +/- 4.7 days, and SAPS II (Simplified Acute Physiology Score) 26 +/- 13 points. The overall mortality was 44 patients (14.5%) with 25 patients (8.3%) dying while on the ICU. RESULTS: The data collection process for TISS took significantly longer than for TISS-28. On the day of admission, the correlation of TISS and TISS-28 was excellent (r(2) = 0.91; p < 0.001). The discriminatory power as assessed by the area under the receiver operating characteristic (ROC) curve was satisfactory for TISS (0.79 +/- 0.04), TISS-28 (0.76 +/- 0.04), and SAPS II (0.77 +/- 0.04) with regard to outcome prediction. Patient-specific costs per TISS-28 point amounted to 36 euros.--and were significantly higher than the 25 euros.--calculated per TISS point. Staff costs (42%) were the most prominent cost-generating factor, and patient-specific costs contributed two thirds to the total ICU costs. There was no association of severity of illness or number of organ failure and costs. Only the length of ICU stay correlated strongly with the costs of the individual patients during the ICU stay (r(2) = 0.79; p < 0.001). CONCLUSION: The faster data collection process as well as the uniformity of the system are strong clinical and scientific advantages of the TISS-28. In addition, TISS-28 is capable of calculating individual costs in an acceptable time frame. Therefore TISS-28 serves as a valuable tool for quality assurance and cost analysis purposes in the medical ICU. PMID- 12647087 TI - [Thrombophilic disorders in venous thromboembolism. The clinical perspective]. AB - BACKGROUND: Treatment of venous thromboembolism should consider the influence of thrombophilic disorders on the risk of recurrence. Diagnostic tests should be guided by their expected relevance to therapy. A competent and straightforward clinical approach to venous thromboembolism requires knowledge of the current data regarding stratification of risk of recurrence in thrombophilic patients. METHODS: Focusing on clinical management, the current literature on thrombophilia is analyzed. Recommendations are given taking into account the risk of first and recurrent thromboembolism, the bleeding risk under anticoagulation, and validated therapy strategies. A rational diagnostic approach in the light of potential therapies is suggested. CONCLUSION: The recurrence risk after triggered venous thromboembolism is low, and anticoagulation does not need to be extended beyond 6 months. Idiopathic or thromboembolism in unusual locations is associated with a higher recurrence risk since underlying thrombophilia is more common in these patients. These cases merit laboratory screening for thrombophilic factors (whose relevance is discussed), and extended anticoagulation is indicated (1 year; in severe thrombophilia several years). In recurrent venous thromboembolism anticoagulation is ideally continued for 4 years, or longer in the absence of complications. PMID- 12647088 TI - [Nephrology--Part 2. Strategies in the immunosuppression after kidney transplantation]. AB - BACKGROUND: As a result of better prevention and treatment of acute rejection, 1 year kidney graft survival exceeds 90% in most centers. By contrast, the rate of attrition over the long-term did not change during the last decades. As current immunosuppressive agents lack specificity (infection, malignant disease), the search for the ideal immunosuppressive agent or drug combinations continues. The goal is to prevent chronic allograft nephropathy, toxicity, infections, malignancies, and metabolic problems. Long-term immunosuppression should be reliable and stable. PURPOSE: This article is aiming to discuss current strategies such as nephrotoxicity-free therapy, avoidance or withdrawal of steroids, therapy of acute rejection, and immunosuppressive therapy in high-risk patients. PMID- 12647089 TI - [Increased myocardial blood flow after spinal cord stimulation in patients with refractory angina pectoris]. AB - BACKGROUND: In spite of great progresses in surgical and catheter interventional techniques there is an increasing number of patients with coronary heart disease not suitable for these conventional treatment strategies. THERAPY: A recent review of the Study Group on the treatment of refractory angina pectoris of the European Society of Cardiology (ESC) recommends spinal cord stimulation (SCS) as first-line therapy. SCS is a well-known and often used therapy for refractory angina in other European countries but not in Germany. The present studies show that SCS is an efficient therapy. By reduction of angina symptoms and a consecutive increase of exercise capacity, the patients experience a great improvement in quality of life. In addition, recent data of our own study suggest a significant decrease in myocardial ischemia in patients under SCS. This might be a direct effect of SCS or due to a better collateralization because of the improved exercise capacity. CONCLUSION: In agreement with the study group of the ESC, we would recommend SCS as first-line therapy for refractory angina pectoris. As a matter of course, conventional treatment strategies should not be replaced by SCS. Hence, a strict evaluation before implanting a SCS device is indispensable. PMID- 12647090 TI - [The journal impact factor as a parameter for the evaluation of researchers and research]. AB - The journal impact factor (IF), which is published annually by the Institute for Scientific Information, (Philadelphia, USA), is meanwhile in widespread use as a scientometric parameter for the evaluation of research and researchers in Germany and other European countries. The present article subjects the IF to critical analysis. It first deals with processes of production, transfer, and use of medical knowledge, because the IF intervenes in these processes on account of its reflexivity. Secondary effects of the IF resulting from its reflexivity are discussed with the focus on the level of the author, the journal and the medical discipline as well as on social knowledge processes in society. In addition, the extent to which the IF is appropriate for evaluating the quality of a specific article, of a journal or of individual and collective research achievements is discussed. The present article calls for (1) research evaluation in accordance with the recommendations of the Deutsche Forschungsgemeinschaft (German Research Council) and of the Arbeitsgemeinschaft der Wissenschaftlichen Medizinischen Fachgesellschaften (Association of the Scientific Medical Societies) and (2) more intensive occupation with and organization of medical knowledge processes. PMID- 12647092 TI - Using weather indices to predict survival of winter wheat in a cool temperate environment. AB - Seven years of winter survival data for winter wheat ( Triticum aestivum L.) were collected on a loam soil located on the Central Experimental Farm at Ottawa, Ontario (45 degrees 23'N, 75 degrees 43'W). The site was low-lying and subject to frequent winter flooding and ice-sheet formation. Two cultivars, a soft white and a hard red winter wheat, were planted in September. Crop establishment was measured in late fall and the percentage survival was measured in April of the following year. Meteorological data, which were available from the nearby weather site, were used to develop a large set of monthly weather indices that were felt to be important for winter survival. The objective of the study was to use genetic selection algorithms and artificial neural networks to select a subset of critical weather factors and topographic features and to model winter survival. The six weather indices selected were the total rain depth for December (mm), the total rain depth for February (mm), the number of days of the month with snow on the ground for January, the extreme minimum observed daily air temperature for March ( degrees C), the number of days of the month with snow on the ground for March, and the number of days of April with a daily maximum air temperature greater than 0 degrees C. It was found 89% of the variation in winter survival could be explained by these six weather indices, the cultivar, elevation and plot location. PMID- 12647091 TI - Weather change and pain: a behavioral animal study of the influences of simulated meteorological changes on chronic pain. PMID- 12647093 TI - Predicting live herbaceous moisture content from a seasonal drought index. AB - In order to provide a relatively simple means of predicting live herbaceous plant moisture content from a readily available meteorological index with an accuracy adequate for fire hazard assessment, the moisture content fluctuations of certain species were correlated with the values of a seasonal drought index based on soil moisture deficiency. The simple linear regression models provided the best fit of the relationship between plant moisture content and the Keetch-Byram drought index (KBDI) values. KBDI predicted with accuracy, for two growing seasons, the moisture content of three annual herbaceous plants ( Piptatherum miliaceum, Parietaria diffusa, Avena sterillis) with shallow rooting systems, typical of the understory vegetation of Pinus brutia forests in the Mediterranean region of Crete, Greece. The greatest aberrations between measured and predicted values of plant moisture content were observed early (May) and late (September) in the growing season, when plant phenology (flushing and withering stages respectively) appears to become the dominant factor in determining plant moisture content regardless of the soil moisture conditions. The KBDI was poorly correlated with the live-needle moisture content of deeply rooted P. brutia trees and modestly with the soil water content of the upper layers. This indicates that the index adequately reflects the moisture condition of the surface soil layers but not the water content deeper in the soil. PMID- 12647094 TI - Effect of uniform and non-uniform skin temperature on thermal exchanges in water in humans. AB - We investigated the effect of uniform (UST) and non-uniform (NUST) skin temperature on thermal exchanges during a 3-h water immersion in five male subjects wearing (NUST) or not wearing (UST) a water-perfused garment. UST was achieved by immersing the nude subject in water up to the neck. For each subject, the water temperature was adjusted to the critical temperature ( T(cw), 31.4 +/- 0.9 degrees C) or 3 degrees C below T(cw) ( T(cw) - 3). NUST was achieved by perfusing different segments of the perfused garment with water of different temperatures. The water temperature of the segment was independently adjusted according to the skin temperature distribution in cold air, the mean skin temperature being the same as the UST. At T(cw) and T(cw) - 3, changes in esophageal and mean skin temperatures were identical in UST and NUST conditions, but the skin temperature of the trunk was higher and that of the limb was lower in the NUST condition. Heat production and the overall skin heat flux at T(cw) were identical in the two conditions, but those at T(cw) - 3 were about 25% lower ( P < 0.05) in NUST than in UST conditions. At T(cw) - 3, the overall tissue insulation was 36% higher ( P < 0.05) in NUST than in UST conditions, mainly because of higher limb insulation. Thermogenesis due to shivering was lower by 62% ( P < 0.05) in NUST than in UST. We conclude that the NUST condition increased tissue insulation and suppressed shivering. This suggests that a high skin temperature of the trunk attenuates shivering in cold water and increases the ability to defend body temperature more economically in cold water. PMID- 12647095 TI - Modeling valley fever (coccidioidomycosis) incidence on the basis of climate conditions. AB - Valley fever (coccidioidomycosis) is a disease endemic to arid regions within the Western Hemisphere, and is caused by a soil-dwelling fungus, Coccidioides immitis. Incidence data for Pima County, reported to the Arizona Department of Health Services as new cases of valley fever, were used to conduct exploratory analyses and develop monthly multivariate models of relationships between valley fever incidence and climate conditions and variability in Pima County, Arizona, USA. Bivariate and compositing analyses conducted during the exploratory portion of the study revealed that antecedent temperature and precipitation in different seasons are important predictors of incidence. These results were used in the selection of candidate variables for multivariate predictive modeling, which was designed to predict deviation from mean incidence on the basis of past, current, and forecast climate conditions. The models were specified using a backward stepwise procedure, and were most sensitive to key predictor variables in the winter season and variables that were time-lagged 1 year or more prior to the month being predicted. Model accuracy was generally moderate ( r(2) values for the monthly models, tested on independent data, ranged from 0.15 to 0.50), and months with high incidence can be predicted more accurately than months with low incidence. PMID- 12647096 TI - A geographical information system model for creating bioclimatic maps - examples from a high, mid-latitude city. AB - This paper presents a method for creating large-scale bioclimatic maps with the aid of a geographical information system, GIS. Meteorological data are linked with geographical information about land use, elevation and distance to the coast, in order to generate spatial distributions of physiological equivalent temperature, PET. The model combines an air temperature map and a wind map in order to create different zones for which the thermal component is to be calculated. The advantage of the model presented is that it uses generally available information about land use, altitude and distance to the coast. Further, the model uses a GIS application, which makes it non-static. Compared to most other models, a wide range of observations are used as input. Few biometeorological studies have been performed in high-latitude areas. This paper presents bioclimatic maps for the Goteborg urban area, in Sweden, for the month of July. The results show large variations in PET during a clear, calm day at 1200 hours (Delta T 13.4 degrees C) and during average conditions in July (Delta T 6.8 degrees C), which gives an indication of the magnitude and the spatial variations within high, midlatitude, urban area in summer. The highest PET values were found in the central built-up areas and the lowest PET values in the coastal and green areas. The model generates valuable information for urban planners and decision makers when planning and constructing new areas for outdoor activities etc. This information is also useful in the fields of health and energy. PMID- 12647098 TI - Effects of male sterility on reproductive traits in gynodioecious plants: a meta analysis. AB - Female fecundity advantage in gynodioecious plants is required for the spread and maintenance of this reproductive system. However, not all reproductive characters show female advantage in all species. We used a meta-analysis to summarise differences between females and hermaphrodites reported from the literature for several reproductive traits. Further we tested three hypotheses, (1) that female plants of species with many ovules produce more seeds per fruit while those with few ovules produce heavier seeds, (2) that females are more pollen limited than hermaphrodites, and (3) that floral sexual size dimorphism is more pronounced in species with few ovules, either because female reproductive success is less limited by pollen availability in such species or because flowers with few ovules require a smaller floral structure to protect the carpels. Overall, females compared to hermaphrodites produced more but smaller flowers, had higher fruit set, higher total seed production, and produced heavier seeds that germinated better. Species with many versus few ovules differed in female advantage for flower size dimorphism, flower number, fruit set and total seed production. However seed size, seed set per fruit and seed germination differences between females and hermaphrodites did not differ significantly between species with few and many ovules. We also found no evidence for differential pollen limitation between females and hermaphrodites. Degree of floral sexual size dimorphism differed significantly between species with few and many ovules. Though pistillate flowers were generally smaller than those of hermaphrodites, species with many ovules showed less difference in flower size between the sexes, suggesting either that the protective role of the perianth constrains the evolution of sexual size dimorphism in species with many ovules or that selection for adequate pollination in species with many ovules impedes the reduction in flower size of females. PMID- 12647099 TI - The carbon charging of pines at the climatic treeline: a global comparison. AB - The carbon charging of pines across the treeline ecotone of three different climatic zones (Mexico 19 degrees N Pinus hartwegii, Swiss Alps 46 degrees N P. cembra and northern Sweden 68 degrees N P. sylvestris) was analyzed, to test whether a low-temperature-driven carbon shortage can explain high-elevation tree limits, and whether the length of the growing season affects the trees' carbon balance. We quantified the concentrations of non-structural carbohydrates (NSC) and lipids (acylglycerols) in all tree organs at three dates during the growing seasons across elevational transects from the upper end of the closed, tall forest (timberline) to the uppermost location where groups of trees > or =3 m in height occur (treeline). Mean ground temperatures during the growing season at the treelines were similar (6.1+/-0.7 degrees C) irrespective of latitude. Across the individual transects, the concentrations of NSC and lipids increased with elevation in all organs. By the end of the growing season, all three species had very similar total mobile carbon (TMC) concentrations at the treeline (ca. 6% TMC in the aboveground dry biomass), suggesting no influence of the length of the growing season on tree carbon charging. At a temperate lowland reference site P. sylvestris reached only ca. 4% TMC in the aboveground dry biomass, with the 2% difference largely explained by higher lipid concentrations of treeline pines. We conclude that carbon availability is unlikely to be the cause of the altitudinal tree limit. It seems rather that low temperatures directly affect sink activity at the treeline, with surplus carbon stored in osmotically inactive compounds. PMID- 12647100 TI - Petiole twisting in the crowns of Psychotria liminesis: implications for light interception and daily carbon gain. AB - We used Y-plant, a computer-based model of crown architecture, to examine the implications of leaf reorientation resulting from petiole bending in Psychotria limonensis (Rubiaceae) seedlings. During this reorientation process, bending of the petioles of lower leaves that are potentially self-shaded by the upper leaves rotates the lamina around the stem's orthotropic axis so that self-shading is reduced. Simulations of daily light capture and assimilation revealed a 66% increase in daily C gain due to reorientation of the leaves as compared to simulations where the leaves remained in their characteristic opposite decussate pattern set by the phyllotaxy. This was due to enhanced carbon (C) gain of the lower leaves because of the reduction of shading by upper developing leaves in the same vertical plane. The light signal for this movement was experimentally examined by placing leaf-shaped filters above already fully expanded leaves and following the resulting shade-avoiding movements. The filters were either neutral density shade cloth that reduced the photon flux density (PFD) but did not alter the red to far red ratio (R:FR) or a film that reduced the PFD equivalently but also reduced the R:FR. Leaf reorientation was much more rapid and complete under the low R:FR as compared to the high R:FR indicating involvement of a phytochrome photosensory system that detected the presence of a shading leaf. Plants in gaps were found to lack a reorientation response indicating that the reorientation is specific to the shaded understory environment. PMID- 12647101 TI - Are lichens active under snow in continental Antarctica? AB - Photosynthetic activity, detected as chlorophyll a fluorescence, was measured for lichens under undisturbed snow in continental Antarctica using fibre optics. The fibre optics had been buried by winter snowfall after being put in place the previous year under snow-free conditions. The fibre optics were fixed in place using specially designed holding devices so that the fibre ends were in close proximity to selected lichens. Several temperature and PPFD (photosynthetic photon flux density) sensors were also installed in or close to the lichens. By attaching a chlorophyll a fluorometer to the previously placed fibre optics it proved possible to measure in vivo potential photosynthetic activity of continental Antarctic lichens under undisturbed snow. The snow cover proved to be a very good insulator for the mosses and lichens but, in contrast to the situation reported for the maritime Antarctic, it retained the severe cold of the winter and prevented early warming. Therefore, the lichens and mosses under snow were kept inactive at subzero temperatures for a prolonged time, even though the external ambient air temperatures would have allowed metabolic activity. The results suggest that the major activity period of the lichens was at the time of final disappearance of the snow and lasted about 10-14 days. The activation of lichens under snow by high air humidity appeared to be very variable and species specific. Xanthoria mawsonii was activated at temperatures below -10 degrees C through absorption of water from high air humidity. Physcia dubia showed some activation at temperatures around -5 degrees C but only became fully activated at thallus temperatures of 0 degrees C through liquid water. Candelariella flava stayed inactive until thallus temperatures close to zero indicated that liquid water had become available. Although the snow cover represented the major water supply for the lichens, lichens only became active for a brief time at or close to the time the snow disappeared. The snow did not provide a protected environment, as reported for alpine habitats, but appeared to limit lichen activity. This provides at least one explanation for the observed negative effect of extended snow cover on lichen growth. PMID- 12647102 TI - Effects of a simple plant morphological mutation on the arthropod community and the impacts of predators on a principal insect herbivore. AB - Plant features that enhance predator effectiveness can be considered extrinsic resistance factors because they result in reduced insect herbivory. In this paper we test the hypothesis that reduced epicuticular wax (EW) in Pisum sativum L. is an extrinsic-resistance factor contributing to field resistance to Acyrthosiphon pisum (Harris). We monitored pea aphid populations in the field on reduced EW and normal EW near isolines of peas for two seasons and confirmed that aphid populations are lower on reduced EW peas than on normal EW peas. We also monitored predators within the canopies of the two pea lines to discover community level patterns in response to differences in EW. We found that while predator numbers were similar between the two lines, there were more syrphids on the normal EW peas, and a trend towards more coccinellids on reduced EW peas. We tested the impact of predators on pea aphids on the two EW lines by monitoring their population levels in cages that excluded predators, and in cages that allowed predators to enter. We found that pea aphid populations were similar on the two EW lines when predators were excluded. When predators were allowed access to the plants, pea aphid populations were reduced more on reduced EW peas than on normal EW peas. We also examined the intrinsic resistance to aphids in reduced EW peas with laboratory dual-choice tests comparing aphid response to reduced EW and normal EW peas, and found that walking, apterous aphids displayed no preference for one pea line over the other. Bioassays to measure growth and fecundity of the pea aphid on the two EW types in the greenhouse and in the field showed that intrinsic rate of increase, and other life table parameters, were not different for aphids on the two lines. Together these results support the hypothesis that reduced EW in peas is a predator-dependent extrinsic resistance factor. Genetically reducing EW bloom in peas and other waxy crop plants might improve the effectiveness of arthropod natural enemies of insect pests. More generally, the results show that a subtle change in plant morphology can substantially influence the impact of predators on insect herbivore populations. The benefit of extrinsic resistance to herbivory conferred by reduced EW may balance any benefits of a prominent EW bloom, thereby sustaining EW polymorphisms in some natural plant populations. PMID- 12647103 TI - Behavioral mechanisms underlie an ant-plant mutualism. AB - Predators can reduce herbivory by consuming herbivores (a consumptive effect) and by altering herbivore behavior, life history, physiology or distribution (non consumptive effects). The non-consumptive, or trait-mediated, effects of predators on prey may have important functions in the dynamics of communities. In a facultative ant-plant mutualism, we investigated whether these non-consumptive effects influenced the host plants of prey. Here, predaceous ants (Forelius pruinosus) consume and disturb a dominant lepidopteran folivore (Bucculatrix thurberiella) of wild cotton plants (Gossypium thurberi). Season-long ant exclusion experiments revealed that ants had a larger proportional effect on damage by B. thurberiella than on caterpillar abundance, a result that suggests ants have a strong non-consumptive effect. Behavioral experiments conducted in two populations over 2 years demonstrated that B. thurberiella caterpillars were substantially less likely to damage wild cotton leaves in the presence of ants due to ant-induced changes in caterpillar behavior. In the absence of ants caterpillars spent more time stationary (potential feeding time) and less time dropping from leaves by a thread of silk than when ants were present. Furthermore, ants altered the spatial distribution of both caterpillars and damage; caterpillars spent relatively more time on the upper surfaces of leaves and caused damage further from the leaf margin in ant exclusion treatments. Both direct encounters with ants and information conveyed when ants walked onto leaves were key events leading to the anti-predator behaviors of caterpillars. This study contributes to a small body of evidence from terrestrial systems demonstrating that the trait-mediated effects of predators can cascade to the host plants of prey. PMID- 12647104 TI - Sex and pollen: the role of males in stabilising a plant-seed eater pollinating mutualism. AB - Some plants are exclusively pollinated by an insect whose larvae feed on their seeds. The net outcome of a single visit for the plant depends on the number of ovules fertilised by the visitor, the number of eggs laid, and the number of seeds eaten by each larva. Unlike other known plant-seed eater pollinating mutualisms, the globeflower-globeflower fly mutualism (Trollius europaeus Chiastocheta spp.) is unique in that not only females but also males visit flowers, and both sexes are potential pollinators. I analysed the relative efficiency of Chiastocheta males versus females in transporting pollen and fertilising globeflower ovules. I show that there is no sex-specific morphological adaptation or behaviour to enhance pollen collection and transportation in Chiastocheta flies, and that males contribute to pollination. However, because of their smaller body size, males transport significantly less pollen than females. Less seeds are produced after a visit from a male than after a visit from a female. A single female visit contributes to about 12% of total seed production, and a single male visit to only 5.4%. Females tend to spend more time inside the flower than males, and the number of ovules fertilised is significantly correlated with the time insects spent inside the closed corolla. The lower efficiency of ovule fertilisation by a male's single visit is compensated for by the higher rate of flower visitation by males: a flower receives about twice as many visits from males as from females during a time unit. The contribution of males to pollination is of major importance with respect to understanding the evolutionary stability of the globeflower globeflower fly mutualism, as males satiate pollen requirement of flowers, masking the antagonistic effect of ovipositing females. PMID- 12647105 TI - Molecular and carbon isotopic composition of leaf wax in vegetation and aerosols in a northern prairie ecosystem. AB - We measured the molecular and carbon isotopic composition of major leaf wax compound classes in northern mixed mesic prairie species (Agropyron smithii, Stipa viridula, Bouteloua gracilis, Tragopogon dubius) and in selected crops (Triticum aestivum, Brassica napus, Hordeum vulgare, Medicago sativa) of southern Alberta and also in aerosols collected 4 m above the prairie canopy. Our aims were to better constrain the wax biosynthetic carbon isotopic fractionation relative to the plant's carbon isotopic discrimination and to quantitatively assess the correspondence between wax composition in vegetation and in boundary layer aerosols. Wax molecular composition of the C(3)prairie species and bulked vegetation was characterized by high abundance of C(28) n-alkanol and C(31) n alkane compounds whereas the C(4) species B. gracilis had several co-dominant n alkanol and n-alkane compounds. Wax molecular composition of crop species differed significantly from that of prairie vegetation and was often dominated by a single compound. Results indicate that leaf wax isotopic composition is quantitatively related to the plant's carbon isotopic discrimination. Although species variations were evident, n-alcohol, n-acid and n-alkane wax compounds were on average depleted in (13)C by approximately 6.0+/-1 per thousand relative to total plant carbon. The magnitude of the depletion in wax delta(13)C was unaffected by environmental factors which altered photosynthetic carbon isotopic discrimination. No consistent difference in the magnitude of wax biosynthetic fractionation was observed between C(3) and C(4) species, indicating that photosynthetic pathway has little influence on the isotopic fractionation of wax during biosynthesis. The isotopic composition of ablated waxes in aerosols collected above the canopy was similar to that of the grassland vegetation but the molecular composition differed significantly and indicated that the source "footprint" of the ablated leaf wax particles we sampled in boundary layer air masses was of a regional or larger spatial scale. PMID- 12647106 TI - Habitat age increases reproduction and nutritional condition in a generalist arthropod predator. AB - We studied the nutritional and reproductive response of Poecilus cupreus (synonymous with Pterostichus cupreus), one of the most abundant carabid beetles in arable land, to the succession in sixteen 1- to 4-year-old wildflower areas. A total of 390 male and 373 female beetles was examined. Each female was dissected and the number of ripe eggs counted. The nutritional state was expressed by a condition factor, which was calculated for each individual based on the observed weight and elytra length of male and female P. cupreus. Carabids in the 1-year old wildflower areas contained significantly less ripe eggs than those from the 4 year-old areas. The condition factor of female and male beetles was significantly lower in 1- than 2- to 4-year-old areas. We examined the influence of habitat parameters (vegetation cover, soil water content, coarse and fine sand, pore volume, habitat size and age, surrounding landscape structure) on the reproductive success and nutritional state of P. cupreus in the 16 wildflower areas. The number of eggs was best explained by habitat age, accounting for 53.4% of the variability. The variation in the condition factor of female and male beetles was best explained by habitat age, which accounted for 73% and 71% of the variation, respectively. Moreover, the beetles' reproductive potential and nutritional condition were significantly associated with vegetation cover, and occasionally also with soil water content. The potential of wildflower areas as a reservoir for the generalist predator P. cupreus was shown to increase with successional age. PMID- 12647107 TI - Infochemical-mediated intraguild interactions among three predatory mites on cassava plants. AB - Carnivorous arthropods exhibit complex intraspecific and interspecific behaviour among themselves when they share the same niche or habitat and food resources. They should simultaneously search for adequate food for themselves and their offspring and in the meantime avoid becoming food for other organisms. This behaviour is of great ecological interest in conditions of low prey availability. We examined by means of an olfactometer, how volatile chemicals from prey patches with conspecific or heterospecific predators might contribute to shaping the structure of predator guilds. To test this, we used the exotic predatory mites Typhlodromalus manihoti and T. aripo, and the native predatory mite Euseius fustis, with Mononychellus tanajoa as the common prey species for the three predatory mite species. We used as odour sources M. tanajoa-infested cassava leaves or apices with or without predators. T. manihoti avoided patches inhabited by the heterospecifics T. aripo and E. fustis or by conspecifics when tested against a patch without predators. Similarly, both T. aripo and E. fustis females avoided patches with con- or heterospecifics when tested against a patch without predators. When one patch contained T. aripo and the other T. manihoti, females of the latter preferred the patch with T. aripo. Thus, T. manihoti is able to discriminate between odours from patches with con- and heterospecifics. Our results show that the three predatory mite species are able to assess prey patch profitability using volatiles. Under natural conditions, particularly when their food sources are scarce, the three predatory mite species might be involved in interspecific and/or intraspecific interactions that can substantially affect population dynamics of the predators and their prey. PMID- 12647108 TI - Biogeography of sponge chemical ecology: comparisons of tropical and temperate defenses. AB - Examples from both marine and terrestrial systems have supported the hypothesis that predation is higher in tropical than in temperate habitats and that, as a consequence, tropical species have evolved more effective defenses to deter predators. Although this hypothesis was first proposed for marine sponges over 25 years ago, our study provides the first experimental test of latitudinal differences in the effectiveness of sponge chemical defenses. We collected 20 common sponge species belonging to 14 genera from tropical Guam and temperate Northeast Spanish coasts (Indo-Pacific and Mediterranean biogeographic areas) and conducted field-based feeding experiments with large and small fish predators in both geographic areas. We use the term global deterrence to describe the deterrent activity of a sponge extract against all of the predators used in our experiments and to test the hypothesis that sponges from Guam are chemically better defended than their Mediterranean counterparts. Sympatric and allopatric deterrence refer to the average deterrent activity of a sponge against sympatric or allopatric predators. All of the sponges investigated in this study showed deterrent properties against some predators. However, 35% of the sponge species were deterrent in at least one but not in all the experiments, supporting the idea that predators can respond to chemical defenses in a species-specific manner. Tropical and temperate sponges have comparable global, sympatric, and allopatric deterrence, suggesting not only that chemical defenses from tropical and temperate sponges are equally strong but also that they are equally effective against sympatric and allopatric predators. Rather than supporting geographic trends in the production of chemical defenses, our data suggest a recurrent selection for chemical defenses in sponges as a general life-history strategy. PMID- 12647109 TI - Nestedness of north-west European land snail ranges as a consequence of differential immigration from Pleistocene glacial refuges. AB - We investigated whether ranges in continental biota are nested. We propose a test for nested subset structure which can detect nestedness even if there are several sets of nested subsets as expected on a larger geographical scale. The test is based on a Monte Carlo simulation with a null model that considers spatial autocorrelation of the occurrences of a taxon. The number of cases in which the occurrences of a species form a subset of the occurrences of another species is used as test statistic. In a case study we show that the ranges of north-west European land snail species are significantly nested. The geographic centres of the sets of nested subsets correlate with glacial refuges. The differential immigration of taxa restricted to southern refuges during the glacials was probably an important mechanism resulting in the observed nestedness of the ranges of the north-west European land snail species. Some species which were more widespread during Pleistocene glacials contribute little to the nested subset pattern and are not nested among themselves. A comparison between groups of species differing in their dispersal abilities indicates that differences in the degree of nestedness are primarily due to differences in the variance of range sizes and not in dispersal abilities. We found a very weak correlation between dispersal ability and the rank of ranges in the sets of nested subsets indicating that nestedness might in part be caused by differential dispersal abilities. We assume that the graded variation of environmental parameters might be more important in generating the nestedness of ranges of north-west European land snail species than their differential dispersal abilities. PMID- 12647110 TI - Factors limiting the intertidal distribution of the mangrove species Xylocarpus granatum. AB - The tree species Xylocarpus granatum is commonly described as occurring in the upper intertidal zone of mangrove forests, but mature trees are occasionally found at lower elevations. In the Utwe River basin, on the Pacific island of Kosrae, we investigated the relative importance of several biotic and abiotic factors that may control the intertidal distribution of X. granatum. Factors we evaluated included differential seed predation across the lower, mid, and upper intertidal zones and seedling responses to salinity, tidal flooding, and shade. Seed predation was 22.4% over the first 34 days and varied little among zones or in gaps versus under the forest canopy. By day 161, there were still no differences in seed mortality, but a significant difference was found in seedling establishment, with much greater establishment in the upper intertidal plots. X. granatum seedlings in a greenhouse experiment exhibited greater growth in freshwater than seedlings in 23 ppt salinity, which is typical of salinity levels found in the mid intertidal zone in our field study sites in Micronesia, where mature X. granatum trees are generally absent. Seedlings grown in 23 ppt salinity, however, exhibited few visible signs of stress associated with patterns in growth. Seedlings grown in a simulated tidal flooding treatment (with 23 ppt salinity) also showed few signs of stress. Growth declined dramatically under 80% shade cloths, but there were few interactions of shading with either 23 ppt salinity or simulated tidal flooding. Differential seed predation is not likely to be the primary factor responsible for the intertidal distribution of X. granatum on Kosrae. However, seedling tolerance of flooding or salinity may be more important, especially relative to a potential contribution to secondary stress mortality. Other factors may ultimately prove to be more critical, such as physiological effects of salinity on seed germination, effects of tides on seed dispersal and rooting, or differential herbivory on seedlings. PMID- 12647111 TI - Czech alien flora and the historical pattern of its formation: what came first to Central Europe? AB - Temporal patterns of immigration to the country were analysed using 668 alien species in the flora of the Czech Republic for which the dates of the first record were available (64.8% of the total number of 1031 so-called neophytes, i.e. aliens introduced after the year 1500). After a period of initial slow increase lasting to the 1840s, the accumulation of neophytes over time could be best fitted by a linear model that explained 97% of the variance. The intensity of floristic research, which varied between periods, did not significantly affect the overall increase in the number of aliens. The effect of species traits on the year of introduction was evaluated, with continent of origin, introduction type (deliberate or accidental), life history, Grime's life strategy, onset of flowering, mode of dispersal and propagule size as explanatory variables. Species of European origin and CSR strategists arrived earlier than those with other origins and strategies. Deliberately introduced species appeared earlier than accidental arrivals, and those cultivated for utilitary reasons on average arrived earlier than ornamentals. Species capable of early flowering were remarkably more prevalent among early newcomers. A separate analysis of accidentally introduced American species also identified life history as a significant predictor of immigration time, with annuals being introduced earlier than biennials and perennials. The data contribute to an understanding of a crucial stage of the invasion process that has received little attention in the literature. The model "early alien" to Central Europe is a European species with a CSR strategy deliberately brought for cultivation as a utilitary plant. Once it escaped from cultivation, its establishment in the wild was favoured by its ability to flower early and, therefore, complete the life cycle. PMID- 12647112 TI - The mud flat anemone-cockle association: mutualism in the intertidal zone? AB - The intertidal cockle Austrovenus stutchburyi exists in a symbiotic relationship with the mud flat anemone Anthopleura aureoradiata, the latter using the shell of buried cockles as the only available hard substrate for attachment. The cockles are also host to a detrimental larval trematode Curtuteria australis that invades the bivalves through the filtration current, and here we demonstrate that the anemones significantly depress the rate by which cockles accumulate parasites in the field. Along the tidal gradient, the relative parasite load of cockles was lowest where anemones were most abundant, and the area occupied by anemones per square meter sediment surface explained 30% of the spatial variation in infection intensity. At a smaller spatial scale, parasite loads were significantly lower (34%) in cockles from patches with than without anemones at the same tidal height. A field experiment manipulating the density of anemones showed that the rate of parasite accumulation in cockles decreased with increasing anemone density, and that the generally positive relationship between infection intensity and cockle size tended to disappear in the presence of anemones. The results suggest that the anemone-cockle symbiosis is a non-obligate mutualistic relationship in which the former is provided with a suitable substrate for attachment whereas the latter obtains protection against parasitic infections. PMID- 12647113 TI - Species decline--but why? Explanations of carabid beetle (Coleoptera, Carabidae) declines in Europe. AB - We investigated some of the causes of ground beetle decline using atlas data from Belgium, Denmark and the Netherlands, countries in which natural environments have all but disappeared. We used ordinal regression to identify characteristics that are significantly correlated with the decline of carabid beetle species over the last 50-100 years, using a stepwise selection procedure to select the optimal model according to the Akaike Information Criterion. The results showed that large-bodied carabid populations have declined more than smaller ones, possibly because of their lower reproductive output and lower powers of dispersal. Habitat specialist populations (i.e. species with small niche breadths) have also decreased more than habitat generalist populations. Species with both long- and short-winged individuals have been less prone to decline than those that are exclusively either short-winged or long-winged. Dimorphic species may survive better in highly altered environments because long-winged individuals are good at dispersing between suitable habitats and short-winged individuals are good at surviving and reproducing in these newly colonised habitats. Finally, populations of large carabids associated with coastal, woodland or riparian habitat types were less prone to decline than populations of large carabids associated with various, open or grassland habitat types. The pattern is reversed for carabid species smaller than 8 mm in size. These results are explained in the context of habitat restoration and destruction in these highly modified western European countries. PMID- 12647114 TI - Influence of the hypogaeic army ant Dorylus (Dichthadia) laevigatus on tropical arthropod communities. AB - The majority of army ant species forage hypogaeically. Due to the difficulties in observing these ants, their potential influence on hypogaeic and epigaeic arthropod communities has not yet been investigated. As the first hypogaeically foraging army ant studied in detail, we attracted Dorylus laevigatus to areas monitored for their arthropod diversity. Here, for the first time, the same sites were sampled before and after an army ant raid. Furthermore, interactions between D. laevigatus and the five most common ground-nesting ant species were noted and their life-history traits compared, allowing first inferences on possible mechanisms of their coexistence. The occurrence of D. laevigatus within a study plot had no evident effect on the number of arthropod taxa or individuals collected with epigaeic and hypogaeic pitfall traps. Likewise, juvenile arthropods, which are less mobile and thus are potentially easier prey for D. laevigatus, showed no differences in their collected numbers before and after the army ant had visited a plot. However, significantly fewer ant species were collected with hypogaeic traps after D. laevigatus had been within the study plots, indicating a possible predation of D. laevigatus especially on two Pseudolasius and one Pheidole species. The five most common ground-foraging ant species demonstrated their ability to avoid, kill, and even prey on the army ant. The reaction of Lophomyrmex bedoti towards D. laevigatus indicated the former to be a potential prey species, while Pachycondyla sp. 2 showed signs of "enemy specification." Odontoponera diversus and O. transversa actively preyed on D. laevigatus, while Pheidologeton affinis fought with D. laevigatus over resources. All ant species could co-occur with D. laevigatus at palm oil baits. Adding to the differences detected in previous studies between D. laevigatus and epigaeically foraging army ant species, the occurrence of this hypogaeic army ant seems to have less devastating effects on arthropod community compositions than those of epigaeically mass raiding species. PMID- 12647115 TI - Swimming ability and behaviour of post-larvae of a temperate marine fish re entrained in the pelagic environment. AB - The degree to which behaviour, vertical movement and horizontal transport, in relation to local hydrodynamics, may facilitate secondary dispersal in the water column was studied in post-larval Sillaginodes punctata in Port Phillip Bay, Australia. S. punctata were captured in shallow seagrass beds and released at the surface in three depth zones (1.5, 3 and 7 m) off-shore at each of two sites to mimic the re-entrainment of fish. The behaviour, depth and position of S. punctata were recorded through time. The direction and speed of local currents were described using an S4 current meter and the movement of drogues. Regardless of site, fish immediately oriented toward the bottom, and into the current after release. In shallow water (1.5 m), 86% of fish swam to the bottom within 2 min of release. At one site, the net horizontal displacement of fish was largely unrelated to the speed and direction of local currents; at a second site, fish could not maintain their position against the current, and the net horizontal displacement was related to the speed and direction of currents. In the intermediate depth zone, wide variability in depths of individual fish through time led to an average depth reached by fish that was between the shallow and deep zones. Based on daily increments in the otoliths, however, this variability was not related significantly to the time since entry of fish into Port Phillip Bay. In the deepest depth zone, 81% of fish remained within 1 m of the surface and their horizontal displacement was significantly related to the direction and speed of currents. Secondary dispersal of post-larval fish in the water column may be facilitated by the behaviour and vertical movements of fish, but only if fish reach deeper water, where their displacement (direction and distance) closely resembles local hydrodynamic regimes. In shallow water, fish behaviour and vertical migration actually reduce the potential for secondary dispersal. PMID- 12647116 TI - Photosynthetic performance and resource utilization of two mangrove species coexisting in a hypersaline scrub forest. AB - In a hypersaline mangrove scrub forest in northern Florida, coexisting trees of Laguncularia racemosa and Avicennia germinans were either fertilized with nitrogen or phosphorus, or not fertilized (controls). We aimed to test whether nutrient additions differentially altered photosynthetic performance and resource utilization in these two species. In control trees, photosynthetic rates were higher in L. racemosa than A. germinans. However, leaf nitrogen concentrations were higher in A. germinans than L. racemosa. Avicennia germinans responded to fertilization with nitrogen by increasing leaf nitrogen concentrations and rates of photosynthesis such that they were equivalent to photosynthesis in L. racemosa. Laguncularia racemosa did not show a response to nitrogen additions. Neither species showed strong responses to phosphorus fertilization. Avicennia germinans had high photosynthetic water-use efficiency (photosynthesis/transpiration), but low photosynthetic nitrogen-use efficiency (photosynthesis/leaf nitrogen). In contrast, L. racemosa had comparatively low photosynthetic water use efficiency and high photosynthetic nitrogen use efficiency. Leaf level characteristics lead us to hypothesize that coexistence of A. germinans and L. racemosa should occur where nitrogen levels are low and salinity is moderate, or at least moderate for some period of the year. PMID- 12647117 TI - Carbon isotope discrimination differences within and between contrasting populations of Encelia farinosa raised under common-environment conditions. AB - Previous studies of the desert shrub Encelia farinosa have shown variation of morpholological and physiological integration that appears to match environmental differences among populations. Such findings led us to ask if there is a genetic basis for such differentiation that may be related to physiological control of intercellular CO(2) concentrations as indicated by carbon isotope discrimination (Delta) values, and if genetic variance for Delta is detectable within populations. Under common environment conditions, Delta values were compared between two populations of E. farinosa from desert regions with contrasting rainfall patterns: Superior, Ariz., a region with high annual rainfall and droughts of short duration, and Oatman, Ariz. a region with lower annual rainfall and longer drought periods. Superior plants had consistently greater mean Delta values than Oatman plants across a broad range of soil water potentials, indicating that there is a genetic basis for Delta variation between these populations. At the intrapopulation level only Oatman plants showed detectable genetic variance of Delta based on: (1) consistent individual-rank values for Delta among soil-drought stages, and (2) evidence of heritable genetic variance for Delta during one drought stage. No genetic variance in Delta was evident for the Superior population. It is hypothesized that the high spatio-temporal heterogeneity of water availability at Oatman may facilitate the maintenance of genetic variance for carbon isotope discrimination within this population. Both the inter- and intra-population level findings suggest that selection associated with rainfall and drought has resulted in genetic divergence of the physiological factors involved in Delta determination for these populations. There appears to be strong differences of water-use and carbon-gain strategies among populations, and broader functional breadth among plants in the habitat of greatest environmental heterogeneity. PMID- 12647118 TI - Covariation in leaf and root traits for native and non-native grasses along an altitudinal gradient in New Zealand. AB - Across 30 grassland sites in New Zealand that ranged from native alpine grasslands to low elevation improved pastures, there were consistent patterns of leaf and root traits and significant differences between native and non-native grasses. Plants of high altitude sites have low N concentrations in both their leaves and roots, have thick leaves and roots, yet no differences in tissue density or photosynthetic water use efficiency when compared to plants of low altitude sites. Both the leaves and roots of the low altitude plants were enriched in (15)N relative to the plants of higher altitude, indicating that the low-N set of traits is associated with a more closed N cycle at high altitude. A second independent set of correlations shows that plants of wetter habitats have lower photosynthetic water use efficiency (more negative partial differential (13)C) and lower leaf and root tissue density than the plants of drier sites. For both leaves and roots, plants of native species consistently had traits associated with lower resource availability: lower N concentrations, denser tissues, more negative partial differential (15)N, and more positive partial differential (13)C than non-native species. If root %N is correlated with root longevity as has been shown in other systems, root longevity may be able to be predicted from simple measurements of leaf %N, though a hysteresis in the relationship between leaf and root N concentrations may make prediction of high longevity roots difficult. PMID- 12647119 TI - Effects of diet and Echinostoma revolutum infection on energy allocation patterns in juvenile Lymnaea elodes snails. AB - Resource allocation strategies may be influenced by both biotic and abiotic factors. The purpose of this study was to investigate the effects of both parasitism and diet quality on the growth, reproduction, and survival of the pond snail, Lymnaea elodes. In addition, we assessed parasite growth and reproduction. High-protein (high diet) or low-protein diets (low diet) were fed to juvenile L. elodes snails that were either exposed or sham-exposed to the castrating trematode, Echinostoma revolutum. Host growth was assessed weekly; reproduction and survival were recorded every 2-3 days. We estimated parasite development as the time to parasite release from the host (patency), and parasite reproduction as the number of larvae shed from infected snails at two time points. Diet and infection status had significant effects on snail growth. Infected snails produced few eggs and tended to grow to larger sizes than uninfected snails regardless of diet. In contrast, exposed-uninfected individuals displayed diet dependent patterns of growth and reproduction. On the high-protein diet, uninfected and exposed-uninfected snails exhibited similar patterns of growth and reproduction, whereas in the low-diet treatment, exposed-uninfected snails exhibited reduced growth and delayed reproduction relative to uninfected individuals. Survival differed among treatments in the latter stages of the study with infected snails exhibiting reduced survival relative to snails from other treatments. Moreover, infected low-diet snails exhibited lower survival than infected high-diet snails. Parasite development and reproduction did not appear to be directly influenced by the quality of host diet. Results from this study suggest that energy allocation patterns are context-dependent in juvenile snails, influenced by parasite exposure and diet quality. Furthermore, parasite reproduction appears to depend more on host size than on the quality of host diet. PMID- 12647120 TI - Seasonal shift from bottom-up to top-down impact in phytophagous insect populations. AB - Although many studies now examine how multiple factors influence the dynamics of herbivore populations, few studies explicitly attempt to document where and when each is important and how they vary and interact. In fact, how temporal variation in top-down (natural enemies) and bottom-up (host plant resources) factors affect herbivore dynamics has been suggested as a particularly important yet poorly understood feature of terrestrial food webs. In this study we examined how temporal changes in predator density (wolf spiders, sheet-web builders, and mirid egg predators) and host-plant resources (plant quality and structural complexity) influence the population dynamics of the dominant phytophagous insects on Atlantic-coast salt marshes, namely Prokelisia planthoppers (Homoptera: Delphacidae). We designed a factorial experiment in meadows of Spartina alterniflora to mimic natural variation in vegetation quality and structure by establishing two levels of plant nutrition (leaf nitrogen content) by fertilization, and two levels of habitat complexity by adding leaf litter (thatch). We then assessed seasonal changes in the strength of bottom-up (plant quality) and top-down (predator) impacts on planthopper populations. Planthopper populations responded positively to increased plant quality treatments in late summer. Despite the greater number of planthopper adults colonizing fertilized Spartina plots compared to unfertilized controls, the offspring of these colonists were much less abundant at the end of the season in fertilized plots, particularly those with thatch. The initial colonization effect was later erased because arthropod predators selectively accumulated in fertilized plots where they inflicted significant mortality on all stages of planthoppers. Predators rapidly colonized fertilized plots and reached high densities well in advance of planthopper colonization, a response we attribute to their rapid aggregation in complex-structured habitats with readily available alternative prey. Our results suggest that plant resources not only mediate the strength of predator impacts on herbivore populations, but they also promote the coupling of predator and prey populations and thus influence when enemy impacts are realized. PMID- 12647121 TI - Marine resources subsidize insular rodent populations in the Gulf of California, Mexico. AB - Inputs of energy and nutrients from one ecosystem may subsidize consumers in adjacent ones, with significant consequences for local communities and food webs. We used stable isotope and faecal pellet analysis to quantify use of ocean derived resources by small mammals on islands in the Gulf of California, Mexico. Rodents were live-trapped on grids originating near shore and extending 125-200 m inland to evaluate the extent to which rodents transport marine nutrients inland, and to determine whether marine foods subsidize island populations, permitting higher densities than would be possible based on terrestrial resources alone. Both faeces and stable carbon and nitrogen isotopes revealed that omnivorous mice (Peromyscus maniculatus) consume ocean-derived prey, including littoral and supralittoral invertebrates, and that their diets differed markedly from those of granivorous rodents (Chaetodipus rudinoris). On a small, seabird roosting island, marine prey were important in the diet of mice regardless of their proximity to shore, underscoring the pervasive influence of the ocean on small islands with relatively large coastline area. On a large island, however, consumption of marine foods declined sharply > or =50 m from shore, which suggests that mice are poor conduits of inland movement of energy and nutrients from the sea. Marine resources seemed to act as subsidies for omnivorous rodents: more P. maniculatus were captured near shore than farther inland and there was an inverse relationship between island area and rodent abundance, suggesting that small islands with large amounts of marine inputs support the highest population densities. Patterns of local and island-wide abundance of P. maniculatus are likely the result of several interacting factors, including frustrated dispersal, competition with C. rudinoris, and the absence of predators. We speculate, however, that the availability of marine resources allows P. maniculatus to reach high densities and to persist on small islands in the Gulf despite low and unpredictable terrestrial productivity. Spatial trophic subsidies thus provide a possible mechanistic explanation for the widely reported inverse relationship between population density and island or habitat area. PMID- 12647122 TI - Estimating the latitudinal origins of migratory birds using hydrogen and sulfur stable isotopes in feathers: influence of marine prey base. AB - Hydrogen stable isotope analysis of feathers is an important tool for estimating the natal or breeding latitudes of nearctic-neotropical migratory birds. This method is based on the latitudinal variation of hydrogen stable isotope ratios in precipitation in North America (deltaD(p)) and the inheritance of this variation in newly formed feathers (deltaD(f)). We hypothesized that the typically strong relationship between deltaD(p) and deltaD(f) would be decoupled in birds that forage in marine food webs because marine waters have relatively high deltaD values compared to deltaD values for local precipitation. Birds that forage on marine prey bases should also have feathers with high delta(34)S values, since delta(34)S values for marine sulfate are generally higher than delta(34)S values in terrestrial systems. To examine this potential marine effect on feather stable isotope ratios, we measured deltaD and delta(34)S in the feathers of nine different species of raptors from both inland and coastal locations across North America. Feathers from coastal bird-eating raptors had consistently higher deltaD and delta(34)S values than feathers from inland birds. Birds that had high delta(34)S values also deviated strongly from the typical relationship between deltaD(p) and deltaD(f). We recommend measuring both sulfur and hydrogen stable isotope ratios in feathers when some members of a migrant population could potentially forage in marine habitats. We suggest using a practical cutoff of delta(34)S >10 per thousand to remove marine-foraging birds from a migrant sample when using stable isotopes of hydrogen to estimate the latitudinal origins of migrants because high deltaD(f) values for marine-foraging birds could potentially distort estimates of origins. PMID- 12647123 TI - El Nino effects on soil seed bank dynamics in north-central Chile. AB - The soil seed bank was monitored in four 75x75 m plots over 6 years (1990-1995) in an arid thorn scrub community in north-central Chile. Sixty-six species were identified. Total seed densities ranged from 2,000 to 42,000/m(2). Average mass of shrub seeds was significantly greater than that of other growth-forms. Between 70 and 90% of the seeds were less than 1 mg, with those in the 0.51-1.00 mg size class being most numerous. Seed densities were highly variable between years as well as within years, but were also closely associated with plant cover patterns and rainfall regime. Higher seed densities were found in wet years, and in samples taken in early summer and early autumn (i.e., after seed set); the lowest seed densities were in late winter (i.e., after annual plant germination). The annual plant species with the highest cover were also the most abundant in the soil seed bank and exhibited the largest seed density fluctuations. In general, seed densities were 5- to 10-fold higher during the 1991-1992 El Nino/southern oscillation (ENSO) years than non-ENSO years, showing the importance of this phenomenon for seed bank replenishment in the arid region of Chile. PMID- 12647124 TI - Food availability affects Osmia pumila (Hymenoptera: Megachilidae) foraging, reproduction, and brood parasitism. AB - Food limitation can reduce reproductive success directly, as well as indirectly, if foraging imposes a risk of predation or parasitism. The solitary bee Osmia pumila suffers brood parasitism by the cleptoparasitic wasp Sapyga centrata, which enters the host nest to oviposit while the female bee is away. I studied foraging and reproduction of O. pumila nesting within cages stocked with rich or sparse floral resources, and the presence or absence of S. centrata to test (1) the response of nesting female O. pumila to food shortages, (2) the response of nesting female O. pumila to the presence of parasites, and (3) whether brood produced under scarce resources are more likely to be parasitized by S. centrata. The rate of brood cell production was significantly lower in cages with sparse floral resources, although females in sparse cages did not produce significantly fewer brood cells overall. Sapyga centrata did not influence the rate of brood cell production, but females exposed to the cleptoparasites had marginally significantly lower reproductive output. Nests in parasite cages had significantly fewer brood cells than those in parasite free cages. The mean duration of foraging bouts made by female O. pumila in sparse cages was not significantly longer than that in rich cages. O. pumila spent less time in the nest between pollen and nectar foraging bouts in sparse cages with S. centrata than those in other cages suggesting that these individuals made more frequent food foraging trips. Despite the weak effects of parasites and bloom density on foraging behavior, O. pumila brood cells experienced a 5-fold higher probability of parasitism by S. centrata in cages with sparse bloom than in those with rich bloom [corrected]. These results support the hypothesis that indirect effects of food scarcity increase O. pumila susceptibility to brood parasitism, although the exact mechanism is not entirely clear yet. PMID- 12647125 TI - Consumption of snowdrop lectin (Galanthus nivalisagglutinin) causes direct effects on adult parasitic wasps. AB - Honeydew is a common sugar-rich excretion of aphids and other phloem-feeding insects and represents the primary sugar in many agricultural systems. When honeydew-producing insects feed on genetically modified plants, the honeydew can contain amounts of the transgene product. Here we address whether this route of exposure poses a risk for non-target insects. Three species of parasitic wasps were selected: i.e. Aphidius colemani, Trichogramma brassicae and Cotesia glomerata, all of which are known to use honeydew as a carbohydrate source in the field. Wasps were fed sucrose solutions with varying concentrations of snowdrop lectin ( Galanthus nivalisagglutinin, GNA), a protein that has been engineered into crops to confer resistance against homopteran pests and that has been detected in honeydew. Parameters evaluated included gustatory response, longevity, fecundity, progeny emergence and sex ratio. While A. colemani and T. brassicae, but not C. glomerata, were able to detect GNA, this gustatory recognition had no effect on the acceptance of a GNA-sucrose solution. In all three species, GNA ingestion reduced parasitoid survival significantly. However, in respect to fecundity, negative effects were observed for T. brassicae but not for A. colemani. The results suggest that the effect of GNA consumption may depend on the specifics of a parasitoid's biology, especially its longevity and its mode of egg maturation. PMID- 12647126 TI - Temperature as a control over ecosystem CO2 fluxes in a high-elevation, subalpine forest. AB - We evaluated the hypothesis that CO(2) uptake by a subalpine, coniferous forest is limited by cool temperature during the growing season. Using the eddy covariance approach we conducted observations of net ecosystem CO(2) exchange (NEE) across two growing seasons. When pooled for the entire growing season during both years, light-saturated net ecosystem CO(2) exchange (NEE(sat)) exhibited a temperature optimum within the range 7-12 degrees C. Ecosystem respiration rate ( R(e)), calculated as the y-intercept of the NEE versus photosynthetic photon flux density (PPFD) relationship, increased with increasing temperature, causing a 15% reduction in net CO(2) uptake capacity for this ecosystem as temperatures increased from typical early season temperatures of 7 degrees C to typical mid-season temperatures of 18 degrees C. The ecosystem quantum yield and the ecosystem PPFD compensation point, which are measures of light-utilization efficiency, were highest during the cool temperatures of the early season, and decreased later in the season at higher temperatures. Branch level measurements revealed that net photosynthesis in all three of the dominant conifer tree species exhibited a temperature optimum near 10 degrees C early in the season and 15 degrees C later in the season. Using path analysis, we statistically isolated temperature as a seasonal variable, and identified the dynamic role that temperature exhibits in controlling ecosystem fluxes early and late in the season. During the spring, an increase in temperature has a positive effect on NEE, because daytime temperatures progress from near freezing to near the photosynthetic temperature optimum, and R(e )values remain low. During the middle of the summer an increase in temperature has a negative effect on NEE, because inhibition of net photosynthesis and increases in R(e). When taken together, the results demonstrate that in this high-elevation forest ecosystem CO(2) uptake is not limited by cool-temperature constraints on photosynthetic processes during the growing-season, as suggested by some previous ecophysiological studies at the branch and needle levels. Rather, it is warm temperatures in the mid-summer, and their effect on ecosystem respiration, that cause the greatest reduction in the potential for forest carbon sequestration. PMID- 12647127 TI - Responses of soil nitrogen dynamics in a Mojave Desert ecosystem to manipulations in soil carbon and nitrogen availability. AB - We investigated the effects of changes in soil C and N availability on N mineralization, nitrification, denitrification, NH(3) volatilization, and soil respiration in the Mojave Desert. Results indicate a C limitation to microbial N cycling. Soils from underneath the canopies of Larrea tridentata (DC.) Cov., Pleuraphis rigida Thurber, and Lycium spp. exhibited higher rates of CO(2 ) flux, lower rates of NH(3) volatilization, and a decrease in inorganic N (NH(4)(+)-N and NO(3)(-)-N) with C addition. In addition to C limitation, soils from plant interspaces also exhibited a N limitation. Soils from all locations had net immobilization of N over the course of a 15-day laboratory incubation. However, soils from interspaces had lower rates of net nitrification and potential denitrification compared to soils from under plant canopies. The response to changes in C availability appears to be a short-term increase in microbial immobilization of inorganic N. Under controlled conditions, and over a longer time period, the effects of C and N availability appear to give way to larger differences due to spatial location. These findings have implications for ecosystems undergoing changes in soil C and N availability due to such processes as desertification, exotic species invasions, or elevated atmospheric CO(2) concentration. PMID- 12647128 TI - Mechanisms behind positive diversity effects on ecosystem functioning: testing the facilitation and interference hypotheses. AB - Little is known about the mechanisms behind positive effects of species richness on ecosystem functioning. In a previous study that showed a positive effect of aquatic detritivore species richness on leaf litter breakdown (process) rates, we proposed that facilitation and release from intra-specific interference were the two most likely mechanisms. To test the interference hypothesis, we performed an experiment using three densities of each of three detritivore species and found varying effects on leaf breakdown rates across species: one species showed no effect, one a positive, marginally insignificant, effect, and a third species showed a significant, positive effect of decreasing density. The density (interference) effect thus partly explained the results from our previous study. The facilitation hypothesis was tested by sequentially introducing and removing two species. We predicted that, if this hypothesis were true, facilitation would be expressed in higher process rates than when replacing with individuals of the same species. We found that process rate per unit biomass did increase when one species was introduced after the other species, while the opposite sequence did not show any increase. Hence, this result was also confirmative of our previous results. Therefore, we conclude that both intra-specific interference and inter specific facilitation may explain the positive effect of species richness observed in our system. Since many species exhibit intra-specific interference that inhibits foraging efficiency, this may be a general mechanism generating effects of species richness per se. If facilitation is unidirectional, or if it involves few species, it is more likely to be species specific with species identities being more important than species richness per se. We conclude that species loss may be expected to have negative consequences on ecosystem functioning if anyspecies is lost, with additional effects in the event of losing "facilitator" species. PMID- 12647129 TI - Ungulate community structure and ecological processes: body size, hoof area and trampling in African savannas. AB - A wide range of bioenergetic, production, life history and ecological traits scale with body size in vertebrates. However, the consequences of differences in community body-size structure for ecological processes have not been explored. We studied the scaling relationships between body mass, shoulder height, hoof area, stride length and daily ranging distance in African ungulates ranging in size from the 5 kg dik-dik to the 5,000 kg African elephant, and the implications of these relationships on the area trampled by single and multispecies herbivore communities of differing structure. Hoof area, shoulder height and stride length were strongly correlated with body mass (Pearson's r >0.98, 0.95 and 0.90, respectively). Hoof area scaled linearly to body mass with a slope of unity, implying that the pressures exerted on the ground per unit area by a small antelope and an elephant are identical. Shoulder height and stride length scaled to body mass with similar slopes of 0.32 and 0.26, respectively; larger herbivores have relatively shorter legs and take relatively shorter steps than small herbivores, and so trample a greater area of ground per unit distance travelled. We compared several real and hypothetical single- and multi-species ungulate communities using exponents of between 0.1 and 0.5 for the body mass to daily ranging distance relationship and found that the estimated area trampled was greater in communities dominated by larger animals. The impacts of large herbivores are not limited to trampling. Questions about the ecological implications of community body-size structure for such variables as foraging and food intake, dung quality and deposition rates, methane production, and daily travelling distances remain clear research priorities. PMID- 12647131 TI - The effect of temporal scale on the outcome of trophic cascade experiments. AB - The outcome of experimental manipulations in ecology should depend on the duration of the experiment. We tested this proposition by examining the results of published trophic cascade experiments in freshwater communities and the results of our own previously unpublished pond experiment. In particular, we analyze the results of 90 published trophic cascade experiments to test the prediction that the outcome of freshwater trophic cascade experiments, in which zooplanktivorous fish are either present or absent, depends on the duration of the experiment. We present evidence that there are quantitative differences in the strength of trophic cascades among different venue types (enclosure, mesocosm, pond, and lake), but that the strength of trophic cascades does not diminish with increasing experiment duration. Despite the large number of studies, there have been few replicated studies of trophic cascades for longer than a summer field season, and none for the time required to estimate the long term result of press perturbations. We therefore present the results of a 4-year study of trophic cascades in experimental ponds, to test the predictions that the addition of the top predator results in a sustained increase in the phytoplankton biomass. We found that, as predicted by our literature review, there was no decline in the strength of the trophic cascade as the experiment progressed. PMID- 12647130 TI - Delta15N values of tropical savanna and monsoon forest species reflect root specialisations and soil nitrogen status. AB - A large number of herbaceous and woody plants from tropical woodland, savanna, and monsoon forest were analysed to determine the impact of environmental factors (nutrient and water availability, fire) and biological factors (microbial associations, systematics) on plant delta(15)N values. Foliar delta(15)N values of herbaceous and woody species were not related to growth form or phenology, but a strong relationship existed between mycorrhizal status and plant delta(15)N. In woodland and savanna, woody species with ectomycorrhizal (ECM) associations and putative N(2)-fixing species with ECM/arbuscular (AM) associations had lowest foliar delta(15)N values (1.0-0.6 per thousand ), AM species had mostly intermediate delta(15)N values (average +0.6 per thousand ), while non mycorrhizal Proteaceae had highest delta(15)N values (+2.9 to +4.1 per thousand ). Similar differences in foliar delta(15)N were observed between AM (average 0.1 and 0.2 per thousand ) and non-mycorrhizal (average +0.8 and +0.3 per thousand ) herbaceous species in woodland and savanna. Leguminous savanna species had significantly higher leaf N contents (1.8-2.5% N) than non-fixing species (0.9 1.2% N) indicating substantial N acquisition via N(2) fixation. Monsoon forest species had similar leaf N contents (average 2.4% N) and positive delta(15)N values (+0.9 to +2.4 per thousand ). Soil nitrification and plant NO(3)(-) use was substantially higher in monsoon forest than in woodland or savanna. In the studied communities, higher soil N content and nitrification rates were associated with more positive soil delta(15)N and plant delta(15)N. In support of this notion, Ficus, a high NO(3)(-) using taxa associated with NO(3)(-) rich sites in the savanna, had the highest delta(15)N values of all AM species in the savanna. delta(15)N of xylem sap was examined as a tool for studying plant delta(15)N relations. delta(15)N of xylem sap varied seasonally and between differently aged Acacia and other savanna species. Plants from annually burnt savanna had significantly higher delta(15)N values compared to plants from less frequently burnt savanna, suggesting that foliar (15)N natural abundance could be used as marker for assessing historic fire regimes. Australian woodland and savanna species had low leaf delta(15)N and N content compared to species from equivalent African communities indicating that Australian biota are the more N depauperate. The largest differences in leaf delta(15)N occurred between the dominant ECM Australian and African savanna (miombo) species, which were depleted and enriched in (15)N, respectively. While the depleted delta(15)N of Australian ECM species are similar to those of previous reports on ECM species in natural plant communities, the (15)N-enriched delta(15)N of African ECM species represent an anomaly. PMID- 12647132 TI - Species richness and regional distribution of myrmecophilous beetles. AB - Four major hypotheses have been put forward to explain local species richness of commensal or parasitic species. The resource distribution hypothesis predicts that regionally widespread host species are able to support higher local species richness of commensals or parasites. On the other hand, the resource size hypothesis predicts that larger hosts can support more species than smaller hosts, and comparably, the resource abundance hypothesis predicts that hosts that offer more resources are able to support more species. Finally, the resource concentration hypothesis predicts that hosts that occur in high-density patches support higher species richness. In this study, we tested the first three of the above hypotheses with myrmecophilous beetles and their host ants. In addition to species richness of myrmecophilous beetles, we also applied the above hypotheses to explain the distribution of the beetles. Our data are exclusively based on an extensive literature survey. Myrmecophilous beetles live in naturally fragmented environments composed of host ant colonies and they are exclusively dependent on ants. We found that the distribution of the host ants and the colony size of the host ants had a positive effect on both the species richness and the distribution of myrmecophilous beetles. In the same way, we found that myrmecophilous beetle species that are generalists, i.e. have more than one host ant species, and thus have more abundant resources, were more widely distributed than specialist species. Thus, we found support for the hypothesis that resource distribution, resource size and resource abundance have an effect on species richness and on the distribution of species. PMID- 12647133 TI - The impact of larval predators and competitors on the morphology and fitness of juvenile treefrogs. AB - Studies of phenotypic plasticity typically focus on traits in single ontogenetic stages. However, plastic responses can be induced in multiple ontogenetic stages and traits induced early in ontogeny may have lasting effects. We examined how gray treefrog larvae altered their morphology in four different larval environments and whether different larval environments affected the survival, growth, development, and morphology of juvenile frogs at metamorphosis. We then reared these juveniles in terrestrial environments under high and low intraspecific competition to determine whether the initial differences in traits at metamorphosis affected subsequent survival and growth, whether the initial phenotypic differences converged over time, and whether competition in the terrestrial environment induced further phenotypic changes. Larval and juvenile environments both affected treefrog traits. Larval predators induced relatively deep tail fins and short bodies, but there was no impact on larval development. In contrast, larval competitors induced relatively short tails and long bodies, reduced larval growth, and slowed larval development. At metamorphosis, larval predators had no effect on juvenile growth or relative morphology while larval competitors produced juveniles that were smaller and possessed relatively shorter limbs and shorter bodies. After 1 month of terrestrial competition among the juvenile frogs, the initial differences in juvenile morphology did not converge. There were no differences in growth due to larval treatment but there were differences in survival. Individuals that experienced low competition as tadpoles experienced near perfect survival as juvenile frogs but individuals that experienced high competition as tadpoles suffered an 18% decrease in survival as juvenile frogs. There were also morphological responses to juvenile competition, but these changes appear to be due, at least in part, to allometric effects. Collectively, these results demonstrate that larval environments can have profound impacts on the traits and fitness of organisms later in ontogeny. PMID- 12647134 TI - Field boundaries as barriers to movement of hover flies (Diptera: Syrphidae) in cultivated land. AB - Field boundaries play an important role as refuges, food sources and corridors for invertebrates and vertebrates, and increasing farmland fragmentation impacts on these functions. However, hedgerows and other structures can also impede dispersal by flying insects. The current work uses the pollen of Phacelia tanacetifolia in hover fly guts as a marker to assess hover fly movement in farm landscapes. In the United Kingdom and New Zealand, Phacelia pollen was found in the guts of Ephisyrphus balteatus and Metasyrphus corollae (United Kingdom) and Melanostoma fasciatum (New Zealand) at distances up to 200 m from the source, when there were no barriers between the flowers and the traps used to catch the flies. The rate of decline over distance in the proportion of flies containing pollen was similar for the two countries. The extent to which four replicated field boundary types impeded hover fly movement was tested using post-and-wire fences, lines of poplars (Populus spp.) with gaps, dense poplars and controls (no potential barriers). Phacelia was planted on one side of each boundary, and along the centre of the control plots. The relative presence of the pollen in flies on both sides of the barriers showed that both types of poplar boundary restricted the movement of the flies, but the fence had no effect. In a separate experiment, gravid females of M. fasciatum were captured at a greater height on a shade-cloth fence than were non-gravid females and males. The implications of this work include the functioning and persistence of metapopulations and the influence of field boundaries on population recovery of beneficial invertebrates following pesticide-induced mortality. If field boundaries contribute to a temporal asynchrony between pest and natural enemy populations, this needs to be considered along with the well-established roles of boundaries as refugia for, and sources of, beneficial arthropods. PMID- 12647135 TI - Hydraulic lift in Acacia tortilis trees on an East African savanna. AB - Recent studies suggest that savanna trees in semi-arid areas can increase understorey plant production. We hypothesized that one of the mechanisms that explains the facilitation between trees and grasses in East African savannas is hydraulic lift (HL). HL in large Acacia tortilis trees was studied during the first 3 months of the dry season during a relatively wet year (1998) and a very dry year (2000). In 1998, we found distinct diel fluctuation in soil water potential (psi(s)), with increasing values during the night and decreasing again the following day. These fluctuations in psi(s )are consistent with other observations of HL and in A. tortilis were found up to 10 m from the tree. In 2000, during a severe drought, psi(s) measurements indicated that HL was largely absent. The finding that HL occurred in wetter years and not in drier years was supported by data obtained on the delta(18)O values in soil, rain and groundwater. The delta(18)O of water extracted from the xylem water of grasses indicated that when they grew near trees they had values similar to those of groundwater. This could be because they either (1) use water from deeper soil layers or (2) use hydraulically lifted water provided by the tree; this was not seen in the same grass species growing outside tree canopies. While our data indicate that HL indeed occurs under Acacia trees, it is also true that psi(s) was consistently lower under trees when compared to outside tree canopies. We believe that this is because tree-grass mixtures take up more water from the upper soil layers than is exuded by the tree each night. This limits the beneficial effect of HL for understorey grasses and suggests that in savannas both facilitation via HL and competition are active processes. The importance of each process may depend upon how wet or dry that particular site or year is. PMID- 12647136 TI - Carotenoid-based plumage coloration of male greenfinches reflects health and immunocompetence. AB - Hypotheses of parasite-mediated sexual selection (PMSS) propose that elaborate male ornaments have evolved due to female preferences. Females would benefit from mating with more ornamental males if males' ornamentation signals their health status and ability to provide parasite resistance genes for the offspring. Carotenoid-based plumage coloration of birds has been hypothesised to honestly reflect an individual's health status due to trade-off in allocation of carotenoids between maintenance and signalling functions. The prediction of this hypothesis, namely that individuals with brighter plumage are able to mount stronger immune responses against novel antigens and reveal generally better health state, was tested in captive male greenfinches (Carduelis chloris). Greenfinches with brighter yellow breast feathers showed stronger humoral immune response against novel antigen (SRBC) while no relationship between plumage coloration and an estimate of cell-mediated immune responsiveness (PHA response) was detected. Elaborately ornamental individuals had better general health state as indicated by the negative correlations between plumage brightness and heterophil haemoconcentration. Consistent with the concept of PMSS, these results suggest that carotenoid-based plumage coloration in greenfinches honestly signals immunocompetence and health status. PMID- 12647138 TI - Root responses and nitrogen acquisition by Artemisia tridentata and Agropyron desertorum following small summer rainfall events. AB - Resources in the Great Basin of western North America often occur in pulses, and plant species must rapidly respond to temporary increases in water and nutrients during the growing season. A field study was conducted to evaluate below ground responses of Artemisia tridentata and Agropyron desertorum, common Great Basin shrub and grass species, respectively, to simulated 5-mm (typical summer rain) and 15-mm (large summer rain) summer rainfall events. The simulated rainfall was labeled with K(15)NO(3) so that timing of plant nitrogen uptake could be monitored. In addition, soil NH(4)(+) and NO(3)(-) concentrations and physiological uptake capacities for NO(3)(-) and NH(4)(+) were determined before and after the rainfall events. Root growth in the top 15 cm of soil was monitored using a minirhizotron system. Surprisingly, there was no difference in the amount of labeled N acquired in response to the two rainfall amounts by either species during the 7-day sample period. However, there were differences between species in the timing of labeled N uptake. The N label was detected in above ground tissue of Agropyron within 1 h of the simulated rainfall events, but not until 24 h after the rainfall in Artemisia. For both Agropyron and Artemisia, root uptake capacity was similarly affected by the 5-mm and 15-mm rainfall. There was, however, a greater increase in uptake capacity for NH(4)(+) than for NO(3)(-), and the 15-mm event resulted in a longer response. No root growth occurred in either species in response to either rainfall event during this 8-day period. The results of this study indicate that these species are capable of utilizing nitrogen pulses following even small summer rainfall events during the most stressful period of the summer and further emphasize the importance of small precipitation events in arid systems. PMID- 12647139 TI - Arrival fat and reproductive performance in a long-distance passerine migrant. AB - Long-distance passerine migrants deposit substantial fat stores to fuel their migratory journey. Many of those migratory birds arrive at their northerly breeding grounds with larger fat stores than were necessary to reach their breeding area. Both male and female American Redstarts ( Setophaga ruticilla) arrived to breed in Michigan's Upper Peninsula with fat, and females arrived with more fat than males in 2 out of 3 years. We test the hypothesis that migrants arriving at the breeding grounds with more body fat have higher reproductive success than birds arriving with little or no fat. Females, and to a lesser extent males, that arrive with fat experience gains in reproductive performance as evidenced by increased clutch size, egg volume, and nestling mass. The results have implications for understanding how events occurring during one phase of the annual cycle influence survival and/or reproductive performance in subsequent phases. PMID- 12647137 TI - Nutritional quality of prebreeding diet influences breeding performance of the Florida scrub-jay. AB - Food supplementation studies of breeding birds have traditionally concentrated on energetic constraints on breeding performance. It is only recently that the nutritional quality of the prebreeding diet has also been considered influential. We examined the importance of specific nutrients in the prebreeding diet of the Florida scrub-jay ( Aphelocoma coerulescens). Birds were provided with one of two supplements (rich in protein and fat or rich in fat only) prior to breeding in 2000 and 2001 and their breeding performance, in relation to unsupplemented (control) birds, was examined. Birds receiving both supplements significantly advanced laying in both years, and increased clutch size in 2000 but not in 2001. Laying date explained variation in clutch size in birds on dietary supplements. Egg mass and volume declined with laying order, irrespective of dietary treatment, but birds on the high fat, high protein diet laid heavier third eggs than controls and this was independent of laying date. Laboratory analysis of 14 abandoned and unhatched eggs revealed that as egg mass increased so did the absolute amount of protein and water while fat content remained relatively fixed. Using these relationships between the masses of egg components and fresh egg mass, we calculated that heavier third eggs laid by birds on high fat and high protein, compared with those laid by controls, contained more water that may be fundamental to chick growth and survival. This is the first demonstration for an avian species that nutritional quality of prebreeding diet can simultaneously influence laying date, clutch size, and egg size and composition. PMID- 12647140 TI - Mechanisms promoting higher growth rate in arctic than in temperate shorebirds. AB - We compared prefledging growth, energy expenditure, and time budgets in the arctic-breeding red knot (Calidris canutus) to those in temperate shorebirds, to investigate how arctic chicks achieve a high growth rate despite energetic difficulties associated with precocial development in a cold climate. Growth rate of knot chicks was very high compared to other, mainly temperate, shorebirds of their size, but strongly correlated with weather-induced and seasonal variation in availability of invertebrate prey. Red knot chicks sought less parental brooding and foraged more at the same mass and temperature than chicks of three temperate shorebird species studied in The Netherlands. Fast growth and high muscular activity in the cold tundra environment led to high energy expenditure, as measured using doubly labelled water: total metabolised energy over the 18-day prefledging period was 89% above an allometric prediction, and among the highest values reported for birds. A comparative simulation model based on our observations and data for temperate shorebird chicks showed that several factors combine to enable red knots to meet these high energy requirements: (1) the greater cold-hardiness of red knot chicks increases time available for foraging; (2) their fast growth further shortens the period in which chicks depend on brooding; and (3) the 24-h daylight increases potential foraging time, though knots apparently did not make full use of this. These mechanisms buffer the loss of foraging time due to increased need for brooding at arctic temperatures, but not enough to satisfy the high energy requirements without invoking (4) a higher foraging intake rate as an explanation. Since surface-active arthropods were not more abundant in our arctic study site than in a temperate grassland, this may be due to easier detection or capture of prey in the tundra. The model also suggested that the cold-hardiness of red knot chicks is critical in allowing them sufficient feeding time during the first week of life. Chicks hatched just after the peak of prey abundance in mid-July, but their food requirements were maximal at older ages, when arthropods were already declining. Snow cover early in the season prevented a better temporal match between chick energy requirements and food availability, and this may enforce selection for rapid growth. PMID- 12647141 TI - Habitat-specific demography: evidence for source-sink population structure in a mammal, the pika. AB - Theory suggests that populations may persist in sink habitats that cannot support replacement-level birth rates. Although it is commonly believed that organisms that can actively select habitat should rarely occur in sinks, the frequency of use of sinks in free-ranging species is not well-documented. We found that a population of American pikas ( Ochotona princeps, Lagomorpha) inhabiting distinct alpine habitats (meadow and snowbed) in Wyoming, USA, had habitat-specific demographic rates that produced a source-sink population structure. Population size increased in both habitats in summer and declined in both habitats in winter, with populations in snowbeds increasing more during summer and decreasing more over winter. Birth rates were consistently higher in meadows and populations in meadows had a consistently higher finite rate of increase (lambda, from life tables) than did those in snowbeds, for which lambda was far below that needed for replacement. Patterns of immigration, population structure, and temporal variation in population size were as expected if meadows were functional sources and snowbeds functional sinks. Patterns of snowmelt differed between habitats, predicted the critical difference in birth rates between habitats, and are a likely primary cause of the differences in habitat-specific birth rates that we observed. This study provides a clear example of source-sink population structure for a mammal. PMID- 12647142 TI - Effects of habitat fragmentation on the fitness of two common wetland species, Carex davalliana and Succisa pratensis. AB - Small habitat size and spatial isolation may cause plant populations to suffer from genetic drift and inbreeding, leading to a reduced fitness of individual plants. We examined the germination, establishment, growth, and reproductive capacity of two characteristic species of mown fen meadows, Carex davalliana, and Succisa pratensis, common in Switzerland. Plants were grown from seeds, which were collected in 18 habitat islands, differing in size and in degree of isolation. We used both common garden and reciprocal transplant experiments to assess effects of habitat fragmentation. In the common garden, plants of Carex originating from small habitat islands yielded 35% less biomass, 30% fewer tillers, and 45% fewer flowering tillers than plants from larger ones. In contrast, plants of Succisa originating from small habitat islands yielded 19% more biomass, 14% more flower heads and 35% more flowers per flower head than plants from larger ones. Moreover, plants of Succisa from small isolated habitats yielded 32% more rosettes than did plants from small connected islands. Reciprocally transplanted plants of Succisa originating from small habitat islands produced 7% more rosettes than plants from larger ones. There was no effect of small habitat size and isolation on germination and establishment of both species in the field. Our results document genetic differences in performance attributable to habitat fragmentation in both species. We suggest that fitness loss in Carex is caused by inbreeding depression, whereas in Succisa the differences in fitness are more likely caused by genetic differentiation. Our study implies that habitat fragmentation affects common habitat-specific species, such as Carex and Succisa, as well as rare ones. PMID- 12647143 TI - Forest mediated light regime linked to amphibian distribution and performance. AB - The vegetation in and around the basins of ephemeral wetlands can greatly affect light environments for aquatic species such as amphibians. We used hemispherical photographs to quantify the light environment in terms of the global site factor (GSF), the proportion of available solar radiation that actually strikes the wetland. We compared GSF to the distribution and performance of two amphibian species (Pseudacris crucifer and Rana sylvatica) within 17 ephemeral wetlands in northeastern Connecticut, USA. We found that P. crucifer is restricted to lighter wetlands (GSF >0.34) and that its abundance is proportional to GSF. By contrast, R. sylvatica is found across the light gradient and its abundance is unrelated to GSF. For both species, GSF is a strong predictor of larval developmental rate. In addition, P. crucifer growth rate is higher in lighter wetlands. Through thermal effects, changes in resources, or other influences, light appears to be an important predictor of the distribution and performance of amphibians. Because the structure of canopies can change rapidly, and because amphibians can be strongly impacted by these changes, vegetation mediated effects on wetland light environments may be critical to understanding the dynamics of amphibian populations within forested biomes. PMID- 12647144 TI - Density-dependent host selection in ectoparasites: an application of isodar theory to fleas parasitizing rodents. AB - Parasites should make the same decisions that every animal makes regarding fitness reward. They can maximize reproductive success by selection of those habitats that guarantee the greatest fitness output. We consider the host population as a habitat of a parasite population. Consequently, hosts (=habitats) that differ quantitatively or qualitatively will support different numbers of parasites. The nature of habitat selection can be detected by isodars, lines along which habitat selection yields equivalent fitness reward. We applied this approach to study host selection of five fleas, each infesting two desert rodents. Xenopsylla conformis, Xenopsylla ramesis, Nosopsyllus iranus theodori and Stenoponia tripectinata medialis parasitize Gerbillus dasyurus and Meriones crassus. Synosternus cleopatrae pyramidis parasitizes Gerbillus andersoni allenbyi and Gerbillus pyramidum. Three fleas ( X. conformis, X. ramesis and S. c. pyramidis) were able to perceive quantitative (amount of the resource; e.g. organic matter in the nest for flea larvae) and/or qualitative (pattern of resource acquisition; e.g. host defensiveness) differences between hosts. Two other fleas did not perceive between-host differences. X. conformis was a density dependent host selector that showed sharp selectivity at low density. X. ramesis and S. c. pyramidis were density-independent host selectors with a direct correspondence of density with habitat quality. N. i. theodori and S. t. medialis were non-selectors with no relationship at all between density and host quality. The results of the application of the isodar theory suggest that ectoparasites, like other animals, behave as if they are able to make choices and decisions that favour environments in which their reproductive benefit is maximized. PMID- 12647145 TI - Exploring the boundary between pollination syndromes: bats and hummingbirds as pollinators of Burmeistera cyclostigmata and B. tenuiflora (Campanulaceae). AB - In this study I documented the degree of specialization in the pollination systems of Burmeistera cyclostigmata and B. tenuiflora (Campanulaceae) to explore the potential role of floral isolation in the diversification of the genus. I asked which floral characteristics are important in specializing on either bat or hummingbird pollination, and whether overlap between these floral syndromes can exist. I examined nocturnal and diurnal pollen deposition, pollinator visitation rates, and single visit effectiveness and related them to intra- and interspecific variation in Burmeistera floral characteristics at Monteverde, Costa Rica. Bats and hummingbirds visited both Burmeistera species, and bats pollinated both species. Owing to differences in floral morphology, however, hummingbirds effectively pollinated only B. tenuiflora. The generalized pollination system of B. tenuiflora demonstrates that there can be overlap in the boundary between ornithophily and chiropterophily, and that nectar production and timing of anthesis do not serve as barriers between these syndromes. The high intraspecific variation in floral color from green to red or purple did not correlate with either nocturnal or diurnal pollen deposition. Degree of flower accessibility did affect pollination; nocturnal pollen deposition significantly decreased as flowers become more obstructed. In Burmeistera, floral morphology and accessibility appear to be the most important floral characteristics for specialization at the boundary between ornithophily and chiropterophily. PMID- 12647146 TI - Why do fig wasps actively pollinate monoecious figs? AB - Active pollination, although rare, has been documented in a few pollination mutualisms. Such behaviour can only evolve if it benefits the pollinator in some way. The wasps that pollinate Ficus inflorescences can be active or passive pollinators. They lay their eggs in fig flowers, so that a proportion of flowers will host a wasp larva instead of a seed. We show in an actively pollinated monoecious fig that lack of pollination does not induce fig abortion or affect wasp offspring size but results in smaller numbers of offspring. Hence, conversely to other active pollination systems, seed formation is not obligatory to sustain developing pollinator larvae; however there is a direct fitness cost to active pollinators not to pollinate. We then compared the locations of eggs and fertilised flowers of three actively pollinated Ficus species and one passively pollinated species. We found that more flowers containing wasp eggs were fertilised in the actively pollinated species relative to those of the passively pollinated one. These results along with comparison with similar studies on dioecious figs, support the hypothesis that active pollination has evolved in fig wasps to ensure that more flowers containing wasp eggs are fertilised as this may increase the chances of successful gall development. The stigmatic platform characterising actively pollinated figs is probably an adaptation to increase pollen dispersion within the fig. PMID- 12647147 TI - Microbial impacts on plant-herbivore interactions: the indirect effects of a birch pathogen on a birch aphid. AB - The role of indirect interactions in structuring communities is becoming increasingly recognised. Plant fungi can bring about changes in plant chemistry which may affect insect herbivores that share the same plant, and hence the two may interact indirectly. This study investigated the indirect effects of a fungal pathogen ( Marssonina betulae) of silver birch ( Betula pendula) on an aphid ( Euceraphis betulae), and the processes underpinning the interaction. There was a strong positive association between natural populations of the aphid and leaves bearing high fungal infection. In choice tests, significantly more aphids settled on leaves inoculated with the fungus than on asymptomatic leaves. Individual aphids reared on inoculated leaves were heavier, possessed longer hind tibiae and displayed enhanced embryo development compared with aphids reared on asymptomatic leaves; population growth rate was also positively correlated with fungal infection when groups of aphids were reared on inoculated branches. Changes in leaf chemistry were associated with fungal infection with inoculated leaves containing higher concentrations of free-amino acids. This may reflect a plant initiated response to fungal attack in which free amino acids from the degradation of mesophyll cells are translocated out of infected leaves via the phloem. These changes in plant chemistry are similar to those occurring during leaf senescence, and are proposed as the mechanistic basis for the positive interaction between the fungus and aphid. PMID- 12647148 TI - Changes in biomass, productivity and decomposition along topographical gradients under different geological conditions in tropical lower montane forests on Mount Kinabalu, Borneo. AB - We have examined how the structure and function of a forest ecosystem change with topography (lower-slope versus ridge) and how the changes are modified by nutrient availability depending on geological substrate (Quaternary and Tertiary sedimentary rocks and ultrabasic rock) in the tropical montane rain forests of Mt. Kinabalu (Borneo) where climate is humid and aseasonal. Reflecting the difference in site age and parent rock, the pool size of soluble-P and inorganic N in topsoil decreased from Quaternary sedimentary >Tertiary sedimentary >ultrabasic rock on the lower-slope, and they decreased from the lower-slope to the ridge on all substrates. Forest structural attributes [stature, above-ground biomass, and leaf area index (LAI)] decreased in the order of Quaternary sedimentary >Tertiary sedimentary >ultrabasic rock in association with soil nutrients on the lower-slopes, and decreased upslope consistently on each of the three substrates. Functional attributes [above-ground net primary productivity (ANPP) and decomposition rate] demonstrated similar patterns to structure. ANPP significantly correlated with LAI among the six sites, while net assimilation rate (ANPP divided by LAI assuming an even productivity between above vs below ground system) was nearly constant. Therefore, ANPP could be explained primarily by LAI. Topographical change in LAI could be explained by leaf mass per area (LMA) combined with stand-level leaf biomass. LMA increased upslope on all substrates in association with the decrease in individual leaf area. Stand-level leaf biomass decreased upslope on all substrates but the Tertiary sedimentary rock. Our study demonstrated that topography and geological substrates interactively affected forest structure and processes. The effect of topography on forest structure and processes was greater on nutrient-rich substrates than on poor substrates, and the effect of geological substrate was greater on lower slopes than on ridges. PMID- 12647149 TI - Nitrogen limitation of growth and nutrient dynamics in a disturbed mangrove forest, Indian River Lagoon, Florida. AB - The objectives of this study were to determine effects of nutrient enrichment on plant growth, nutrient dynamics, and photosynthesis in a disturbed mangrove forest in an abandoned mosquito impoundment in Florida. Impounding altered the hydrology and soil chemistry of the site. In 1997, we established a factorial experiment along a tree-height gradient with three zones, i.e., fringe, transition, dwarf, and three fertilizer treatment levels, i.e., nitrogen (N), phosphorus (P), control, in Mosquito Impoundment 23 on the eastern side of Indian River. Transects traversed the forest perpendicular to the shoreline, from a Rhizophora mangle-dominated fringe through an Avicennia germinans stand of intermediate height, and into a scrub or dwarf stand of A. germinans in the hinterland. Growth rates increased significantly in response to N fertilization. Our growth data indicated that this site is N-limited along the tree-height gradient. After 2 years of N addition, dwarf trees resembled vigorously growing saplings. Addition of N also affected internal dynamics of N and P and caused increases in rates of photosynthesis. These findings contrast with results for a R. mangle-dominated forest in Belize where the fringe is N-limited, but the dwarf zone is P-limited and the transition zone is co-limited by N and P. This study demonstrated that patterns of nutrient limitation in mangrove ecosystems are complex, that not all processes respond similarly to the same nutrient, and that similar habitats are not limited by the same nutrient when different mangrove forests are compared. PMID- 12647150 TI - Helpful habitant or pernicious passenger: interactions between an infaunal bivalve, an epifaunal hydroid and three potential predators. AB - The importance of positive interspecific interactions within physically stressful habitats has received increased attention from community ecologists. The exposed sandy beach is an example of a physically rigorous environment where biological interactions have long been considered insignificant. We examined the interaction between the infaunal clam, Donax variabilis, and the hydroid, Lovenella gracilis, on exposed sandy beaches in North Carolina. Epibiotic occupation of Donax by hydroids has been repeatedly observed on ocean beaches but rarely investigated. By providing a stable substrate for attachment, the clam facilitates the persistence of the hydroid in the intertidal beach; however, benefits or costs experienced by the host as a result of this association are unknown. By exposing clams with and without hydroid colonies to multiple types of clam predators, we tested the effectiveness of the hydroid, which possesses stinging nematocysts, in defending its host. The hydroid defended the clam against one common predator, the Florida pompano ( Trachinotus carolinus). Against speckled crabs ( Arenaeus cribrarius) and ghost crabs ( Ocypode quadrata), however, the hydroid offered no protection for its host and instead facilitated predation. The epibiotic hydroid, which projects above the surface of the sand, allowed the crabs to more readily detect clams below the surface. In the field, we evaluated the effect of the hydroid on the tidally synchronized migrations and burrowing speed of the clam. The hydroid, which can form large colonies on the posterior end of the clam, had no effect on Donax burrowing speed, but did reduce the speed of transport of clams by wave swash. Depending on relative predation pressure, the occupation of D. variabilis by L. gracilis can alternately be characterized as beneficial or detrimental to the host. PMID- 12647151 TI - Effects of Formica ants on soil fauna-results from a short-term exclusion and a long-term natural experiment. AB - Wood ants (Formica spp.) were hypothesised to affect the composition and greatly reduce the abundance of large-sized soil fauna by predation. This was tested in two ways. Firstly, a 4-year-long experimental study was carried out in a mixed forest. Five ant-free 1.3-m(2) plots were created by fenced exclosures within an ant territory. Five nearby plots had fences with entrances for the ants. In addition, five non-fenced control plots were selected. Soil fauna (e.g. Coleoptera, Diptera larvae, Collembola and Araneae) was sampled during the summers of 1997-2000. The soil fauna was affected by the exclosures but there was no detectable effect of ants on the soil fauna. Secondly, soil fauna was studied within a large-scale natural experiment in which the long-term (30 years) effects of red wood ants could be assessed inside and outside ant territories. This long term natural experiment revealed no significant effects of ants on the abundance or composition of soil fauna. The results from the two studies indicate that the effects of wood ants on soil fauna are fairly small. The hypothesis that wood ants are key-stone predators on soil fauna could, thus, not be supported. PMID- 12647152 TI - Positive interactions between migrant and resident birds: testing the heterospecific attraction hypothesis. AB - We experimentally tested the conditions where heterospecific attraction is more likely to occur. The heterospecific attraction hypothesis predicts that colonizing or migrant individuals use the presence of resident species as a cue for profitable breeding sites. In other words, increasing resident densities will result in increased migrant densities until the costs of interspecific competition override the benefits of heterospecific attraction. The experiment consisted of a reference and a manipulation year. In the reference year, resident titmice were permitted to breed at intermediate densities whilst in the manipulation year, resident densities were manipulated in nine study plots. Three treatments were performed as low, intermediate and high resident densities and migrant density responses were measured in both years. Relative between-year migrant and resident densities were analyzed by regression analysis. Migrant foliage gleaning guild densities responded linearly and positively, as did densities of habitat generalists, in particular Chaffinch ( Fringilla coelebs),. The ground-foraging guild did not show a response. This study provides support for predictions of the heterospecific attraction hypothesis and suggests that information on habitat quality with reference to both food availability and safe breeding sites are important in using heterospecifics as cues. Based on Chaffinch response data, artificially increased resident densities were not high enough for competitive effects between residents and migrants to decrease heterospecific attraction. It seems unlikely that in northern environments natural resident densities will reach high levels where competitive effects would occur, therefore heterospecific attraction will always be beneficial. This study again shows the importance of heterospecific attraction in migrant habitat selection and as a process promoting species diversity in northern breeding bird assemblages. PMID- 12647153 TI - Seasonal variation in the niche, habitat availability and population fluctuations of a bivoltine thermophilous insect near its range margin. AB - We investigated the niche requirements of the summer and autumn/spring generations of the bivoltine butterfly, Polyommatus bellargusRott., and their implications for population dynamics at sites occurring near its northern range margin. The larvae of this species are sedentary, and the turf height and shelter of Hippocrepis comosa foodplants selected for egg-laying accurately predict larval distributions within United Kingdom (UK) sites. We found a significant shift between the plants used for egg-laying in each generation, with the niche occupied by summer-feeding larvae being broader and different to the autumn one. Measurements of soil temperature confirmed that the short, sheltered foodplants selected by ovipositing females in autumn placed the autumn/spring-feeding generation of larvae in the warmest available microclimates within sites. In late spring, egg-laying females avoided the hottest spots but extended egg-laying into taller, less sheltered (relatively cool) turf where the microclimate was similar to that experienced by autumn/spring-feeding larvae. Using each generations' definition of niche requirement, we analysed surveys of foodplant populations available on 24 UK sites for P. bellargus, and estimated that nearly twice as many plants were available to the summer-feeding larvae compared to those feeding in the autumn. Annual adult population counts match these seasonal differences in site carrying capacity; first generation counts (from autumn-laid eggs) were generally half as abundant as in the second generation, and more variable. These results suggest that the seasonal cycle of niche switches represents an annual (autumn-spring) bottleneck for populations of this butterfly at its northern range margin. Under climate warming we predict that the inter-generational difference in niche availability, carrying capacity and population size will be reduced. We recommend revised management requirements for this threatened species under current and predicted climates in northern Europe. PMID- 12647156 TI - Compensation for herbivory by Cucumis sativus through increased photosynthetic capacity and efficiency. AB - Herbivory is an important selective pressure in the life history of most plant species, as it usually results in reduced plant fitness. In some situations, however, plants are able to compensate for the resources lost to herbivory and do not suffer any reduction in growth or reproduction after attack. We examined the ability of Lebanese cucumber (Cucumis sativus) to compensate for both pre flowering and during-flowering foliar herbivory through increased photosynthetic efficiency and capacity. Plants that were damaged before flowering were able to compensate, in terms of vegetative biomass and fruit production for up to 80% leaf area loss. Plants that were damaged during the flowering period were less able to compensate and fruit production declined with increasing herbivory. Damaged plants had higher photosynthetic efficiency and capacity, and dissipated less light energy as heat. Herbivore-damaged plants may be induced to use a greater proportion of the absorbed light energy for photosynthesis as a result of altered carbohydrate source-sink relationships. PMID- 12647154 TI - Thyme and isolation for the Sinai baton blue butterfly (Pseudophilotes sinaicus). AB - The distribution of the narrowly endemic butterfly Pseudophilotes sinaicus (Lycaenidae) was studied. Potential habitat within its range was first located and then the quality of that habitat assessed. Degree of shelter, diversity of plant species, and resource area of an individual food plant (Thymus decussatus) all affected habitat quality and together were used to develop an index of habitat suitability applicable to each site. The butterfly's distribution was then studied within the identified network of suitable habitat patches: isolated patches with a small resource area were least likely to contain butterflies. Population size in a patch (as opposed merely to patch occupancy) was affected by resource area and the quality of habitat within that patch. Metapopulation processes and variation in habitat quality therefore appear to combine to describe the distribution of patches occupied by P. sinaicus and their population sizes. This finding provides insights into some of the processes operating on an endemic species throughout its geographical range and has important implications for the conservation of this rare butterfly. PMID- 12647157 TI - Impact of fertilisation on phenol content and growth rate of Cladina stellaris: a test of the carbon-nutrient balance hypothesis. AB - Responses of concentrations of usnic (UA) and perlatolic (PA) acids and the relative growth rate (RGR) of a mat-forming lichen, Cladina stellaris, to enhanced N and P input were studied in a fertilisation experiment. It was predicted on the basis of carbon-nutrient balance (CNB) hypothesis that the concentrations of these phenolics would decline and the growth rate increase in response to increased nutrient uptake. The concentration of UA showed a convex response pattern to increased N input whereas the concentration of PA was non responsive. An ecologically realistic, "moderate", N treatment clearly lowered the level of UA both with and without the P application. Applying P alone caused a significant increase in the level of UA. The RGR of C. stellaris did not respond to nutrient addition. The results indicate that even though the CNB hypothesis may be applicable in explaining concentrations of lichen secondary metabolites, it may be applied under a relatively narrow set of conditions. Especially inherited constraints in the growth of lichen fungi may seriously limit the responsiveness of lichens to short-time changes in the availability of resources. These limitations may also apply to other perennials adapted to nutrient-poor conditions. PMID- 12647158 TI - Phenotypic variation in smooth softshell turtles (Apalone mutica) from eggs incubated in constant versus fluctuating temperatures. AB - Temperatures experienced during embryonic development elicit well-documented phenotypic variation in embryonic and neonatal animals. Most research, however, has only considered the effects of constant temperatures, even though developmental temperatures in natural settings fluctuate considerably on a daily and seasonal basis. A laboratory study of 15 clutches of smooth softshell turtles (Apalone mutica) was conducted to explicitly examine the influence of thermal variance on phenotypic variation. Holding mean temperature constant and eliminating substrate moisture effects permitted a clear assessment of the impact of thermal variance on hatching success, incubation length, hatchling body size, swimming speed, and righting time. Incubation length and swimming speed varied significantly among temperature treatments. Both traits tended to increase with increasing thermal variance during embryonic development. Clutch significantly affected all traits examined, except righting time, even after accounting for the effects of initial egg mass. These results highlight the importance of accounting for the impact of both thermal mean and variance on phenotypic variation. The findings also strengthen the increasing recognition of maternal clutch effects as critical factors influencing phenotypic variation in neonatal animals. PMID- 12647159 TI - Adaptive reproductive variation along a pollution gradient in a wolf spider. AB - When populations are exposed to environmental pollutants, growth and reproduction might be strongly reduced due to an increased detoxification effort. Sublethal metal pollution is therefore to be expected to cause the same selection pressure as a low resource habitat and might alter the reproductive strategy. Optimality models of life history theory predict that when resource availability is reduced, growth and reproductive output are reduced and that the release of fewer but larger propagules will be favoured. This was tested by applying a life history model to reproductive trait measurements in six populations of the wolf spider Pirata piraticus in which the assumptions of the model are satisfied. Internal Cd, Cu and Zn body burden were strongly correlated with each other, and differed strongly between the populations, indicating consistently differing metal exposure at the different sites. Pb levels were extremely variable within each population and did not differ between the populations. Females from populations with high concentrations of the first three heavy metals showed a strongly reduced reproductive output and fecundity, indicating a high reduction in resource availability due to detoxification processes. Egg size in contrast was negatively correlated with fecundity and reproductive output and as a consequence positively related with internal metal burden. Our results are thus in strong agreement with the predictions of the optimality models and confirm the benefits of a larger propagule size when resource availability is reduced. PMID- 12647160 TI - Effects of brood parasitism on host reproductive success: evidence from larval interactions among dung beetles. AB - This paper investigates the effect of brood parasitism in a dung beetle assemblage in an arid region of Spain. The study was conducted during the spring season (March-May 1994-1998) using mesh cylinders buried into the ground, filled with sand and with sheep dung on top. We quantified the proportion of nests containing larvae of parasitic beetles and their effect on host larvae survival. Experiments on the effect of parasitic larvae on host-larvae survival were conducted by placing scarab brood masses (raised from captive scarabs in the laboratory) in containers with and without aphodiid larvae. During the spring, dung desiccation is rapid, preventing aphodiids nesting in the dung, and forcing these species to adopt brood parasitism as a nesting strategy. Parasitic aphodiids were found in 12-47% of scarab nests of three species. The incidence of brood parasitization was positively related with the number of brood masses contained in the nests, being also higher in the most abundant species. Field data and experiments showed that brood parasites significantly reduced host larvae survival from 74.8% in non-parasitized nests to 8.8% in parasitized nests. Because different rates of nest parasitization and mortality were caused by parasites, brood parasitism had a differential effect on different host species. Thus, brood parasitism constitutes an important mortality factor reducing the reproductive success of the host species and potentially affecting the beetle abundance in the area. PMID- 12647161 TI - Spatial responses of two herbivore groups to a geometrid larva on mountain birch. AB - Direct or plant-mediated interactions between herbivores may modify their spatial distribution among and within plants. In this study, we examined the effect of a leaf-chewing geometrid, the autumnal moth (Epirrita autumnata), on two different herbivore groups, leaf rolling Deporaus betulae weevils and Eriocrania spp. leafminers, both feeding on mountain birch (Betula pubescens ssp. czerepanovii). The exact locations of herbivores within tree canopies were mapped during three successive summers. In the first 2 years, some trees were artificially colonized by eggs of the autumnal moth to induce both rapid and delayed resistance in the foliage. The natural infection levels of the pathogenic rust fungus (Melampsoridium betulinum), potentially involved in species interactions, were also recorded. At the level of the whole tree, the density of D. betulae leaf rolls was lower in trees infested by the autumnal moth in the same year. However, the feeding locations within trees were partly segregated: D. betulae favoured shadier branches, while E. autumnata preferred the sunny parts of the canopy. The autumnal moth did not affect current- or following-year density of leafminers at the tree or branch level. Trees infected by rust had fewer leafminers in the same summer than noninfected trees. There were no interaction effects between defoliation by the autumnal moth and rust infection, and no delayed effects on the abundance of other herbivores the following year. Taken together, these findings suggest that the autumnal moth has a negative, partially plant-mediated impact on D. betulae, and can reduce the extent of current-year defoliation caused by D. betulae. This may be beneficial for the mountain birch, since the greater part of D. betulae damage occurs around or after the end of the larval period of the autumnal moth, which may be a critical time for tree recovery after moth outbreaks. PMID- 12647162 TI - Catalpa bignonioides alters extrafloral nectar production after herbivory and attracts ant bodyguards. AB - Inducible anti-herbivore defenses are found within many plant taxa, but there are fewer examples of inducible indirect defenses that incorporate the third trophic level. This study links caterpillar foraging, herbivore-induced changes in extrafloral nectar production, and the attraction of ants to vulnerable leaves and plants. Catalpa bignonioides Walter (Bignoniaceae) uses extrafloral nectar to attract ant (Forelius pruinosus(Roger)) bodyguards in response to Ceratomia catalpae (Boisduval)(Lepidoptera: Sphingidae) herbivory. Ant density per leaf increased with the sugar content of extrafloral nectar excreted by sampled leaves, suggesting that increased nectar production could attract or retain beneficial arthropods. The masses of sucrose, fructose, glucose and all three sugars combined in the extrafloral nectar increased two- to three-fold on attacked leaves within 36 h of the experimental addition of caterpillars. Production rates for neighboring non-attacked leaves and non-attacked leaves on adjacent plants did not differ over the same time period. Ant attendance at caterpillar-attacked leaves increased two- to three-fold within 24 h of herbivory, relative to attendance at neighboring, undamaged leaves. These attacked leaves attracted the fewest ants prior to the onset of herbivory, suggesting the specialist caterpillar may avoid or be excluded from leaves with more bodyguards. The removal of leaf tissue with scissors did not alter ant attendance at damaged leaves. Mean ant attendance per leaf on attacked plants increased 6- to 10-fold after caterpillar introduction, relative to adjacent unattacked plants. The plant's biotic defense thus operates at two scales; the number of bodyguards (ant workers) on the plant increases after attack, and this increased workforce is biased towards attacked leaves within plants. Fewer caterpillars remained on plants that attracted greater numbers of ants, suggesting these bodyguards benefit the plant. PMID- 12647163 TI - Foraging ecology of bison at the landscape and plant community levels: the applicability of energy maximization principles. AB - Predictions of animal distribution and resource use require multi-scale consideration because animals can use different sets of selection criteria at different scales. We investigated whether patterns of distribution and resource use by free-ranging bison (Bison bison) in Prince Albert National Park, Saskatchewan, follow rules of energy maximization that hold across multiple scales. Optimality theory predicts specialization on Carex atherodes and frequency-independent selection among plant species; that is, local variation in C. atherodes biomass should not influence diet choice but only the time spent in individual patches. The summer use of resources within meadows was closely related to energy maximization principles. C. atherodes dominated the diet of bison, was selected in all meadows, and diet choice was frequency-independent among meadows in the bison range. In winter, diet was still dominated by C. atherodes, but frequency-dependent selection of Scolochloa festucacea and the relative use of Cirsium arvense were inconsistent with theoretical predictions. At a larger spatial scale, however, the probability of meadow use was not positively related to the abundance of Carex atherodes. During summer and winter, general landscape features within the daily radius of bison (2 km in summer and 1.3 km in winter), together with abiotic characteristics of meadows, had the major influence on the probability of meadow utilization. Our study suggests that bison distribution and resource use are influenced by abiotic and biotic factors which vary in importance at different spatio-temporal scales. PMID- 12647164 TI - Use of novel pollen species by specialist and generalist solitary bees (Hymenoptera: Megachilidae). AB - If trade-offs between flexibility to use a range of host species and efficiency on a limited set underlie the evolution of diet breadth, one resulting prediction is that specialists ought to be more restricted than generalists in their ability to use novel resource species. I used foraging tests and feeding trials to compare the ability of a generalist and a specialist solitary mason bee species to collect and develop on two pollen species that are not normally used in natural populations (novel pollens). Osmia lignaria (Hymenoptera: Megachilidae) is a generalist pollen feeder; O. californica, is more specialized. Adults of the specialist were more limited in use of novel hosts, but only in some contexts. Both bee species refused to collect one novel pollen. The specialist accepted a second novel pollen only when it was presented along with its normal pollen, whereas the generalist collected novel pollen whether presented alone or with normal pollen. Surprisingly, larvae of the specialist were more flexible than were generalists. The specialist grew well on mixtures of normal and novel pollen species, in some cases better than on its normal host alone. Larvae of the generalist grew more poorly on all diets containing novel pollens than on their normal host. Data on these two species of bees suggest that specialization by itself need not reduce flexibility on novel hosts. The findings also provide information about mechanisms of specialization in bees. Similar to some folivores, specific cues of the pollen host and the bee's interpretation of these contribute, along with foraging economics, to pollen choice by adults. The ability of the larvae to cope with specific components of one pollen species need not interfere with its ability to use others. PMID- 12647165 TI - Carbon and nitrogen transfer from a desert stream to riparian predators. AB - Adult aquatic insects emerging from streams may be a significant source of energy for terrestrial predators inhabiting riparian zones. In this study, we use natural abundance delta(13)C and delta(15)N values and an isotopic (15)N tracer addition to quantify the flow of carbon and nitrogen from aquatic to terrestrial food webs via emerging aquatic insects. We continuously dripped labeled (15)N NH(4) for 6 weeks into Sycamore Creek, a Sonoran desert stream in the Tonto National Forest (central Arizona) and traced the flow of tracer (15)N from the stream into spiders living in the riparian zone. After correcting for natural abundance delta(15)N, we used isotopic mixing models to calculate the proportion of (15)N from emerging aquatic insects incorporated into spider biomass. Natural abundance delta(13)C values indicate that orb-web weaving spiders inhabiting riparian vegetation along the stream channel obtain almost 100% of their carbon from instream sources, whereas ground-dwelling hunting spiders obtain on average 68% of their carbon from instream sources. During the 6-week period of the (15)N tracer addition, orb-web weaving spiders obtained on average 39% of their nitrogen from emerging aquatic insects, whereas spider species hunting on the ground obtained on average 25% of their nitrogen from emerging aquatic insects. To determine if stream subsidies might be influencing the spatial distribution of terrestrial predators, we measured the biomass, abundance and diversity of spiders along a gradient from the active stream channel to a distance of 50 m into the upland using pitfall traps and timed sweep net samples. Spider abundance, biomass and richness were highest within the active stream channel but decreased more than three-fold 25 m from the wetted stream margin. Changes in structural complexity of vegetation, ground cover or terrestrial prey abundance could not account for patterns in spider distributions, however nutrient and energy subsidies from the stream could explain elevated spider numbers and richness within the active stream channel and riparian zone of Sycamore Creek. PMID- 12647166 TI - Seed dispersal by wind, birds, and bats between Philippine montane rainforest and successional vegetation. AB - In the moist Neotropics, vertebrate frugivores have a much greater role in the dispersal of forest and successional woody plants than wind, and bats rather than birds play the dominant role in dispersing early successional species. I investigated whether these patterns also occurred in a Philippine montane rainforest and adjacent successional vegetation. I also asked whether seed mass was related to probability of dispersal between habitats. A greater number of woody species and stems in the forest produced vertebrate-dispersed seeds than wind-dispersed seeds. Although input of forest seeds into the successional area was dominated by vertebrate-dispersed seeds in terms of species richness, wind dispersed seeds landed in densities 15 times higher. Frugivorous birds dispersed more forest seeds and species into the successional area than bats, and more successional seeds and species into the forest. As expected, seed input declined with distance from source habitat. Low input of forest seeds into the successional area at the farthest distance sampled, 40 m from forest edge, particularly for vertebrate-dispersed seeds, suggests very limited dispersal out of forest even into a habitat in which woody successional vegetation provides perches and fruit resources. For species of vertebrate-dispersed successional seeds, probability of dispersal into forest declined significantly with seed mass. PMID- 12647167 TI - Redundancy among three herbivorous insects across an experimental current velocity gradient. AB - We conducted streamside experiments to determine if the ability of herbivorous insects to remove algal periphyton varies with local current velocity. We used two mayfly species (Baetis bicaudatusand Drunella grandis) and one caddisfly species (Glossosoma verdona), which differ from one another in body morphology and mobility. Periphyton was grown for 30 days on ceramic tiles in constant velocity to create similar initial forage conditions for grazers. Tiles were transferred to three velocity regimes characteristic of the natural streambed: slow (3-5 cm s(-1)), medium (15-20 cm s(-1)) and fast (32-41 cm s(-1)). Four grazer treatments (Baetis, Drunella, and Glossosoma alone, and all species combined) were repeated for each velocity treatment to isolate the effect of local current on grazer ability to crop periphyton. Grazers differed in their abilities to remove periphyton across current treatments. Glossosoma removed significantly (P<0.05) more periphyton at fast versus either slow or medium velocities; Baetis showed a similar (but non-significant) trend; and, Drunella always removed about 75% of periphyton, irrespective of current. At fast current, periphyton removal was equivalent among the species. At medium current, Drunella removed significantly more than both Baetis and Glossosoma, whereas at slow current, Drunella removed more than Baetis, which removed more than Glossosoma. Periphyton removal under the combined three-grazer treatment was similar qualitatively to the combined effects of individual grazers. More periphyton tended to be removed as current increased, with the fast versus slow contrast showing marginal significance (P=0.10). Under all current regimes, the quantity of periphyton removed did not differ from the null model expectation of simple additive effects among individual grazers (i.e., no facilitation or inhibition). These experiments show that for some species, herbivory varies with current, which suggests that the herbivore "function" of cropping periphyton may vary with the environmental context of local current. Under some local velocities, however, different herbivore species "function" similarly and are potentially redundant with respect to periphytic removal. In naturally heterogeneous streams characterized by sharp gradients in local current velocity, we expect current dependent species interactions to be common and at least partially contribute to intra-guild co-existence of species. PMID- 12647168 TI - Tadpoles' responses to risk of fish introduction. AB - The introduction of predatory species, such as fish, in amphibian breeding sites is one of the many likely causes of amphibian population decline. The existence of inducible or constitutive (permanent) defences is expected to temper the lethal effects of fish on tadpoles. According to current theories on the evolution of phenotypic plasticity, the amphibian species that occur in habitats that are mostly or often fish-free were expected to display inducible defences, while the species that reproduce mainly in fish ponds ought to present constitutive defences. The tested species were Rana dalmatina, Hyla arborea and R. ridibunda, respectively. As expected, R. ridibunda tadpoles were less vulnerable to fish predation than the other species. Nevertheless, all three species exhibited morphological responses in the presence of fish. Their increases in tail area and decreases in body depth were similar. However, behaviour was only modified in R. dalmatina, which exhibited lower activity rates and higher refuge use in the presence of fish cues. Such lower activity rates were related to lower growth and developmental rates. While not detected in this experiment, the existence of inducible defences in the three species suggests the existence of costs of anti-predator defence. Those costs may have delayed effects so that increasing the frequency of fish ponds may exacerbate the impact of delayed costs whatever the species, suggesting that the dynamics of species that are not fish-adverse could also suffer from the introduction of fish. PMID- 12647169 TI - Morphological variation of the predatory cladoceran Leptodora kindtii in relation to prey characteristics. AB - The responses of invertebrate predators to changes in the morphology of their prey, and especially the responses for induced defences, are largely unexplored, compared with the vast amount of data on predator-induced defences. This study demonstrates that the size of the feeding basket, the anatomical structure used to capture prey by the predaceous freshwater cladoceran Leptodora kindtii, shows significant allometric changes with the average body size of the prey (herbivorous cladocerans) in six lakes of northeastern Poland. Prey density influences the basket size only in adult Leptodora individuals, whereas induced defences such as helmets of Daphniasp. seem to have no effect on Leptodora's morphology. In a feeding experiment it is shown that the increase in the feeding basket enables Leptodora to capture larger prey. The seasonal pattern of morphological change, and its measurable benefit suggest that the observed morphological variability of Leptodora is phenotypic plasticity. PMID- 12647170 TI - Effects of experimental food supplementation on movements of juvenile northern goshawks (Accipiter gentilis atricapillus). AB - Food availability is thought to strongly influence the leaving phase of natal dispersal in animals with condition-dependent dispersal. We conducted a food supplementation experiment to determine the influence of extra food on the onset of dispersal and early dispersal movement patterns of 42 radio-tagged northern goshawks (Accipiter gentilis atricapillus) from 28 broods in north-central New Mexico during 1992 and 1993. We randomly assigned half of the broods each year as treatments and the other half as controls. Treatment broods were given supplemental food from hatching (late April) until mid-October. Control broods received the same visitation rate but no food. Birds were located approximately every 2 days from fledging until mid-October in 1992 and 1993. Timing of fledging and independence were not affected by the treatment. However, extra food significantly influenced post-fledging movements of juvenile goshawks. During the late fledgling-dependency period (>65 days of age until independence) control birds were located in the natal area (<2 km from nest tree) more frequently than supplemented birds. This pattern reversed after independence (approximately 82 days of age) when supplemented birds were located more frequently in the natal area than controls. After independence the control birds were never located in the natal area and by the end of September in both years the controls had all left the study area (study area boundaries were approximately 25 km from nest tree). Supplemented birds were never located outside of the study area for the duration of the experiment. We conclude that the control birds dispersed out of the study area and the supplemented birds remained. Since the experimentally fed juveniles remained near a known food source and the controls did not, our study demonstrates that food availability influences at least the first 4 months of post-fledging movement patterns in this population. These results also suggest individuals base dispersal decisions on knowledge of their environment at a local scale, which can influence juvenile recruitment. PMID- 12647172 TI - Functional convergence in plant responses to the environment. AB - Much comparative ecophysiological research has focused on contrasting species specific behavior or ecological strategies with regard to regulation of basic physiological processes such as transpiration, photosynthesis and growth, leading to an emphasis on divergence rather than convergence in plant functioning. This review highlights selected examples in which substantial functional convergence among taxonomically, phylogenetically and architecturally diverse species has been revealed by applying appropriate scaling factors and identifying universal constraints or trade-offs. Recent empirical and theoretical scaling models emphasize the strong role that plant size, architecture, allometry and chemistry play in constraining functional traits related to water and carbon economy and growth. Taken together, the findings summarized here strongly suggest that there are a limited number of physiological solutions to a given problem of plant adaptation to the environment. Comparative ecophysiological studies will therefore benefit from consideration of the constraints that plant anatomical, structural and chemical attributes place on physiological functioning. PMID- 12647173 TI - Sex-ratio dependent execution of queens in polygynous colonies of the ant Formica exsecta. AB - Formica exsecta has become an important model system for studying intraspecific variation in sex ratios. Patterns of sex allocation in polygynous (multiple queen per nest) populations of F. exsecta are generally consistent with the queen replenishment hypothesis. This hypothesis states that colonies produce gynes (reproductive females) in order to increase queen number and enhance colony survival and/or productivity when the number of resident queens is low. However, the small proportion of colonies that raise gynes produce more than necessary for simple queen replenishment. It has been hypothesized that excess production of gynes may occur to reduce the frequency of accepting foreign unrelated gynes into the colony when workers cannot distinguish nestmate from non-nestmate queens. This explanation for excess gynes requires weak or no aggression between non nestmates and is expected to lead to the selective execution of new queens by colonies that do not invest in the production of gynes. Experimental studies where gynes were introduced into natal and foreign colonies indeed suggested that polygynous populations of F. exsecta have a poor nestmate recognition system. Although gynes were significantly more likely to be accepted in their parental colony compared to another foreign female-producing colony, the difference was small. Moreover, encounters between workers from different colonies within the population showed very little aggression and were no more aggressive than encounters between nestmates, again suggesting a weak capacity for nestmate recognition. Our experiment also showed that colonies that produced only males executed most of the gynes that were experimentally introduced into the colony, whereas female-producing colonies accepted most gynes. This is consistent with ants using a simple rule of thumb to decrease parasitism by unrelated queens, whereby colonies selectively destroy gynes whenever gynes are not produced in the colonies. PMID- 12647174 TI - Seed size and germination response: a relationship for fire-following plant species exposed to thermal shock. AB - Thermal shock is well known to be an important stimulus for the germination of soil-stored seeds in fire-prone plant communities. Nevertheless, while the overall germination response of different species is known to vary, the interaction between seed size and germination to a range of thermal-shock temperatures is poorly understood. This interaction may be important in regulating post-fire plant community establishment, since larger seeds are able to emerge from deeper within the soil profile than smaller seeds, and are therefore likely to be insulated against high above-ground temperatures by a deeper soil covering. In this experiment we examined how germination of eight co occurring Western Australian fire-followers was influenced by thermal shock, and whether germination was significantly correlated with seed size. We found that small-seeded species not only showed enhanced germination at higher temperatures, but that their ability to germinate at higher temperatures was also greater than that displayed by larger-seeded species. These findings suggest that while seed size may be a useful general predictor of post-fire recruitment success, under different fire regimes the interaction between seed size, maximum seedling emergence depth, and the ability to withstand different thermal-shock temperatures is complex and may confound recent predictive models. PMID- 12647175 TI - Kinetics of photoacclimation in corals. AB - Traditional models describing the relationship between photosynthesis (P) and irradiance (I) do not account for photoacclimation to short-term variation in irradiance. Here we develop and test a model that predicts the rate of photosynthesis under fluctuating irradiances at the scale of days to weeks. Using oxygen respirometry, we measured the rates of change in the P-I model parameters P(max) (maximum rate of gross photosynthesis) and I(k) (sub-saturation irradiance) of the photo-symbiotic coral Turbinaria mesenterina (Lamarck) following large and small increases and decreases in growth irradiance. We analyse the behaviour of the dynamic P-I model in turbid-water conditions using a dataset of 3-month continuous irradiance as the input variable. In response to upward or downward changes in experimental growth irradiance, I(k) values decreased or increased exponentially, reaching new and stable levels within 5-10 days. I(k) responded 4 times stronger than P(max) to changes in growth irradiance. The kinetics of I(k) did not show hysteresis, and changed in similar ways when irradiance was increased or decreased in small or large amounts. This suggests that mechanisms associated with photo-protection during increases in irradiance, and the maximisation of photosynthetic efficiency during decreases in irradiance, are equally potent. On the scale of months, the dynamic P-I model did not predict higher rates of photosynthesis than the static P-I model, but buffered the variation in photosynthesis during periods of reduced irradiance. Fourier analysis indicated that the kinetics of I(k) closely matches the main periodicities in daily irradiance (1-2 weeks). The recorded kinetics of photoacclimation in the Turbinaria-zooxanthella symbiosis is comparable to that of free-living phytoplankton and faster than that of higher plants. PMID- 12647176 TI - Interactive effects of nitrogen and irradiance on sustained xanthophyll cycle engagement in Eucalyptus nitens leaves during winter. AB - Eucalyptus nitens is a species that is adapted to low temperature. This study examines xanthophyll-cycle engagement in E. nitens seedlings exposed to cold induced photoinhibitory conditions under different levels of irradiance and nutrient status. Xanthophyll-cycle pool size indicated an increased requirement for light energy dissipation under high irradiance and low nutrient status. Greater sensitivity to photoinhibition of non-shaded seedlings indicated that sustained xanthophyll-cycle engagement may occur in response to damaged chlorophyll. Within irradiance treatments, fertilised seedlings had higher photochemical efficiency and faster recovery from photoinhibition than unfertilised seedlings. These results demonstrate that fertilised compared to unfertilised seedlings can utilise a greater proportion of incident light under cold temperature conditions PMID- 12647177 TI - Axial and radial water transport and internal water storage in tropical forest canopy trees. AB - Heat and stable isotope tracers were used to study axial and radial water transport in relation to sapwood anatomical characteristics and internal water storage in four canopy tree species of a seasonally dry tropical forest in Panama. Anatomical characteristics of the wood and radial profiles of sap flow were measured at the base, upper trunk, and crown of a single individual of Anacardium excelsum, Ficus insipida, Schefflera morototoni, and Cordia alliodora during two consecutive dry seasons. Vessel lumen diameter and vessel density did not exhibit a consistent trend axially from the base of the stem to the base of the crown. However, lumen diameter decreased sharply from the base of the crown to the terminal branches. The ratio of vessel lumen area to sapwood cross sectional area was consistently higher at the base of the crown than at the base of the trunk in A. excelsum, F. insipida and C. alliodora, but no axial trend was apparent in S. morototoni. Radial profiles of the preceding wood anatomical characteristics varied according to species and the height at which the wood samples were obtained. Radial profiles of sap flux density measured with thermal dissipation sensors of variable length near the base of the crown were highly correlated with radial profiles of specific hydraulic conductivity (k(s)) calculated from xylem anatomical characteristics. The relationship between sap flux density and k(s) was species-independent. Deuterium oxide (D(2)O) injected into the base of the trunk of the four study trees was detected in the water transpired from the upper crown after only 1 day in the 26-m-tall C. alliodora tree, 2 days in the 28-m-tall F. insipida tree, 3 days in the 38-m-tall A. excelsum tree, and 5 days in the 22-m-tall S. morototoni tree. Radial transport of injected D(2)O was detected in A. excelsum, F. insipida and S. morototoni, but not C. alliodora. The rate of axial D(2)O transport, a surrogate for maximum sap velocity, was positively correlated with the predicted sapwood k(s) and with tree height normalized by the relative diurnal water storage capacity. Residence times for the disappearance of the D(2)O tracer in transpired water ranged from 2 days in C. alliodora to 22 days in A. excelsum and were positively correlated with a normalized index of diurnal water storage capacity. Capacitive exchange of water between stem storage compartments and the transpiration stream thus had a profound influence on apparent rates of axial water transport, the magnitude of radial water movement, and the retention time in the tree of water taken up by the roots. The inverse relationship between internal water exchange capacity and k(s) was consistent with a trade-off contributing to stability of leaf water status through highly efficient water transport at one extreme and release of stored water at the other extreme. PMID- 12647178 TI - Water source utilization by Pinus jeffreyi and Arctostaphylos patula on thin soils over bedrock. AB - Stable isotopes were used to evaluate water sources for co-occurring Jeffrey pine (Pinus jeffreyi Grev & Balf.) and greenleaf manzanita (Arctostaphylos patula Greene) in the southern Sierra Nevada, California, where soils averaged only 75 cm thick but were underlain by up to 5 m of weathered granitic bedrock. Soils and underlying weathered bedrock were sampled three times during both the 1997 and 1998 growing seasons, in 25 cm increments, from 0 to 400 cm or until hard bedrock was reached, and plant stem tissue was sampled simultaneously. Extracted water from the soil/bedrock substrate and plant tissue was analyzed for delta(18)O and/or deltaD, and depth of water source was determined by inference in conjunction with moisture status of the substrate. Water source utilization over the growing seasons for both plants generally followed a pattern similar to that observed for water depletion. Predominant water use was initially from the surface soils. Progressively deeper water sources, including weathered bedrock to a depth of several meters, were exploited as the season progressed and the overlying substrate was depleted of moisture. Early in the growing season, stable isotope values were slightly lower for pine than for manzanita (e.g., average deltaD in June 1997 was -81 per thousand for pine and -77 per thousand for manzanita), and suggest that the functional rooting depth for pine may have been slightly greater than for manzanita. In September 1997, manzanita deltaD values averaged -57 per thousand while pine values averaged -85 per thousand, indicating that manzanita opportunistically utilized summer precipitation while pine used more dependable bedrock water. In 1998, soils remained moist through July due to a late snowfall. Unlike the previous year, pine and manzanita deltaD values were not significantly different in mid- and late-growing season, and both plants exploited bedrock-derived water as soil water was depleted. Water held within bedrock was essential for meeting plant transpirational requirements over the summer drought. PMID- 12647179 TI - Direct nocturnal water transfer from oaks to their mycorrhizal symbionts during severe soil drying. AB - Symbiotic mycorrhizal fungi play an important role in the absorption of soil nutrients and water by most plants. It has been suggested that hydraulically lifted water might maintain the integrity of the external mycorrhizal mycelium during drought. We tested this hypothesis in the obligately mycorrhizal species, coast live oak (Quercus agrifolia), using a microcosm system that separated the effects of hydraulic lift in roots from those in the external mycelium. Mycorrhizal oak seedlings were established in microcosms comprising three discrete compartments for (1) upper roots, (2) tap roots, and (3) external fungal mycelium. Eight months after planting, a drought treatment was initiated: irrigation to the upper root and fungal chambers was terminated and only irrigation to the taproot compartment was maintained. After 3, 12, 30, 50, 70 and 80 days of drought, tracers were injected into the taproot compartment at dusk. At dawn the following morning, mycorrhizal hyphae (EM and AM) and spores (AM) in upper root and fungal compartments were extensively labeled with the tracers. In contrast, no labeling was observed when tracers were injected into the taproot compartment during daytime. Nocturnal water translocation from plant to mycorrhizal fungi occurred in association with hydraulic lift. Saprotrophic/parasitic fungi in the microcosms were not labeled, suggesting a direct water transfer from plants to their mycorrhizal mutualists and not to other fungi in the soil. Even after prolonged drought (70-80 days), mycorrhizal hyphae persisted in soils with water potential values as low as -20 MPa. Maintaining mycorrhizal activity through direct water translocation could potentially improve the nutrient status of deep-rooted plants during periods when the fertile upper soil is dry. PMID- 12647181 TI - A double test of the parasite manipulation hypothesis in a burrowing bivalve. AB - The parasite manipulation hypothesis predicts that parasites should be selected to manipulate host behaviour to facilitate transmission to the next host. The bivalve Macoma balthica burrows less deep when parasitized by the trematode Parvatrema affinis. Shallow burrowing increases the likelihood of ingestion by birds, their final hosts, and therefore this has been interpreted as manipulation by the parasite. When unparasitized, M. balthica displays seasonal changes in burrowing depth, becoming less accessible to predators in winter. If shallow burrowing of parasitized individuals is due to direct manipulation by the parasite, the availability of parasitized individuals should be high throughout the year, or at least especially in the season when most birds are present and potential transmission rates are highest. We compared burrowing depths of parasitized and unparasitized individuals in a single population during seven consecutive years. Parasitized individuals showed reduced burrowing depths but, in contrast to the prediction, the effect of parasites on availability to predators was smallest, not largest, in the season with the highest bird numbers. The parasite P. affinis competes for energy with the host, and M. balthica with low energy stores are known to reduce depth of burrowing. When we included size corrected somatic ash-free dry mass (as an estimate of the energy stores) in our statistical analysis, the effect of infection on burrowing depth disappeared. Thus the effect of infection on burrowing depth is likely to be an unavoidable, indirect effect of the channelling of energy towards the parasite, causing the starving individual to take greater risks in the acquisition of food. Since both the seasonal pattern and the magnitude of increased availability of parasitized individuals are inadequate, the increased exposure of parasitized M. balthica to the final host does not seem to represent an example of adaptive host manipulation by the parasite. PMID- 12647182 TI - Root cooperation in a clonal plant: connected strawberries segregate roots. AB - The ability to selectively avoid competition with members of the same clone should be highly advantageous but has not been demonstrated in plants. We found that physical connection between plants in a clone of the wild strawberry Fragaria chiloensis induced them to segregate their roots, significantly increasing clonal performance. Such increase in performance was not found when plants were grown in containers that artificially divided their rooting zones. There was no effect of connection in a different clone of F. chiloensis with a lower degree of carbon transport between connected plants, suggesting that the mechanism for root segregation depended upon transport of a signal through the strawberry runners. We suggest that clonal integration allows some clones to coordinate below-ground resource foraging with other clone members, thus exhibiting a type of root cooperation. PMID- 12647183 TI - Does prior breeding experience improve reproductive success in collared flycatcher females? AB - In order to assess the importance of breeding experience in shaping age-specific patterns of reproduction an experiment on collared flycatchers (Ficedula albicollis) was performed. Experimental females were restrained from reproduction by destroying their nests after prolonged incubation. Unmanipulated females were used as controls. Only young, first time breeders were considered. A year after manipulation, experimental (inexperienced) and control (experienced) females were compared for a number of breeding traits. They did not differ with respect to laying date, clutch size, number of fledglings and number of local recruits. However, fledglings from broods of inexperienced females were significantly lighter and had shorter tarsi. This suggests that prior experience of feeding young may indeed influence the future reproductive performance in altricial bird species such as the collared flycatcher. PMID- 12647184 TI - Elevated CO2 lowers relative and absolute herbivore density across all species of a scrub-oak forest. AB - The unabated increase in global atmospheric CO(2) is expected to induce physiological changes in plants, including reduced foliar nitrogen, which are likely to affect herbivore densities. This study employs a field-based CO(2 )enrichment experiment at Kennedy Space Center, Florida, to examine plant herbivore (insect) interactions inside eight open-topped chambers with elevated CO(2) (710 ppm) and eight control chambers with ambient CO(2). In elevated CO(2) we found decreased herbivore densities per 100 leaves, especially of leaf miners, across all five plant species we examined: the oak trees Quercus myrtifolia, Q. geminata, and Q. chapmanii, the nitrogen-fixing vine Galactia elliottii and the shrub Vaccinium myrsinites. Both direct and indirect effects of lowered plant nitrogen may influence this decrease in herbivore densities. Direct effects of lowered nitrogen resulted in increased host-plant related death and an increase in compensatory feeding: per capita herbivore leaf consumption in elevated CO(2) was higher than in ambient CO(2). Indirectly, compensatory feeding may have prolonged herbivore development and increased exposure to natural enemies. For all leaf miners we examined, mortality from natural enemies increased in elevated CO(2). These increases in host-plant induced mortality and in attack rates by natural enemies decreased leaf miner survivorship, causing a reduction in leaf miner density per 100 leaves. Despite increased leaf production in elevated CO(2) from the carbon fertilization effect, absolute herbivore abundance per chamber was also reduced in elevated CO(2). Because insects cause premature leaf abscission, we also thought that leaf abscission would be decreased in elevated CO(2). However, for all plant species, leaf abscission was increased in elevated CO(2), suggesting a direct effect of CO(2) on leaf abscission that outweighs the indirect effects of reduced insect densities on leaf abscission. PMID- 12647185 TI - Elimination of plant toxins by herbivorous woodrats: revisiting an explanation for dietary specialization in mammalian herbivores. AB - Constraints on rates of detoxification and elimination of plant toxins are thought to be responsible for limiting dietary specialization in mammalian herbivores. This hypothesis, known as the detoxification limitations hypothesis, suggests that most mammalian herbivores are generalists to avoid overdosing on toxins from a single plant species. The hypothesis also predicts that the few mammalian specialists that exist should have adaptations for rapid detoxification and elimination of plant secondary compounds. We took a pharmacological approach to test whether specialists eliminate toxins from the bloodstream faster than generalists. We compared elimination rate and total exposure of alpha-pinene in closely related dietary specialist and generalist woodrats, Neotoma stephensi and N. albigula, respectively. Animals were orally gavaged with alpha-pinene, a plant secondary compound present in the natural diets of both woodrat species. We collected venous blood at 3, 6, 10, 15, and 20 min post-ingestion of alpha pinene. Blood was analyzed for alpha-pinene concentration using gas chromatography. We found that specialist and generalist woodrats did not differ in elimination rates of alpha-pinene. However, specialists had lower exposure levels of alpha-pinene than generalists due to lower initial delivery of alpha pinene to the general circulation. The levels of alpha-pinene detected in the bloodstream of specialists were 4.7-5.3x lower over all time intervals than generalists. Thus, specialists encounter a functionally lower dose of toxin than generalists. We suggest that the lower exposure level of specialist woodrats may be due to mechanisms in the gut that decrease toxin absorption. Regardless of mechanism, lower exposure to plant toxins may allow specialists to forage on diets with high toxin concentrations thereby facilitating dietary specialization. PMID- 12647186 TI - Effects of elevated carbon dioxide and ozone on the phytochemistry of aspen and performance of an herbivore. AB - The purpose of this study was to assess the independent and interactive effects of CO(2), O(3), and plant genotype on the foliar quality of a deciduous tree and the performance of a herbivorous insect. Two trembling aspen (Populus tremuloides Michaux) genotypes differing in response to CO(2) and O(3) were grown at the Aspen FACE (Free Air CO(2) Enrichment) site located in northern Wisconsin, USA. Trees were exposed to one of four atmospheric treatments: ambient air (control), elevated carbon dioxide (+CO(2); 560 microl/l), elevated ozone (+O(3); ambient x1.5), and elevated CO(2)+O(3). We measured the effects of CO(2) and O(3) on aspen phytochemistry and on performance of forest tent caterpillar (Malacosoma disstria Hubner) larvae. CO(2) and O(3) treatments influenced foliar quality for both genotypes, with the most notable effects being that elevated CO(2) reduced nitrogen and increased tremulacin levels, whereas elevated O(3) increased early season nitrogen and reduced tremulacin levels, relative to controls. With respect to insects, the +CO(2) treatment had little or no effect on larval performance. Larval performance improved in the +O(3) treatment, but this response was negated by the addition of elevated CO(2) (i.e., +CO(2)+O(3) treatment). We conclude that tent caterpillars will have the greatest impact on aspen under current CO(2) and high O(3) levels, due to increases in insect performance and decreases in tree growth, whereas tent caterpillars will have the least impact on aspen under high CO(2) and low O(3) levels, due to moderate changes in insect performance and increases in tree growth. PMID- 12647188 TI - Adult beetle grazing induces willow trichome defence against subsequent larval feeding. AB - Induced increases in trichome density to grazing by herbivores have been suggested to increase plant resistance to future herbivore attack. In this paper we present results which show that plants of Salix cinerea L. respond to adult leaf beetle (Phratora vulgatissima L.) grazing by developing new leaves with an increased trichome density. The same plants are usually attacked again later in the season when the next generation of larvae hatches on the plants. The effect of the induced response was studied by comparing larval growth and feeding on newly developed leaves of plants exposed to different defoliation treatments. Larvae on plants previously exposed to adult grazing consumed less total leaf area and showed more dispersed feeding than larvae on plants protected from previous grazing. Larvae on plants exposed to mechanical defoliation responded intermediately. These results corresponded to the increased trichome density of defoliated plants. However, we found this larval response only in whole plant tests--when reared on single, excised leaves in petri dishes, larvae in all treatments behaved similarly. This discrepancy between the on-plant experiment and that in petri dishes highlights how experimental design may alter the conclusion of a study. We suggest that the induced response to adult grazing may act as a defence against subsequent larval feeding. PMID- 12647187 TI - Why does the larval integument of some sawfly species disrupt so easily? The harmful hemolymph hypothesis. AB - The larvae of several sawfly species belonging to the Tenthredinidae (Hymenoptera) have such a low mechanical resistance in the integument that slight mechanical damage to the integument is enough to provoke the release of hemolymph at a given spot. We quantified this phenomenon, which we call "easy bleeding", by measuring the pressure needed to pierce dissected sawfly integument. We also investigated the feeding deterrance of ethanolic extracts of the hemolymph by laboratory bioassays using Myrmica rubra ant workers. These traits, integument resistance and hemolymph deterrence, were inversely related, considering 22 tenthredinid species. A negative correlation was obtained by only taking into account the species of one tenthredinid tribe, namely the Phymatocerini (nine species studied). Our results support the "harmful hemolymph hypothesis" that we present here and that assumes a functional link between these morphological and chemical traits, jointly acting as a chemical defense strategy. We suspect hemolymph deterrence to be often due to sequestration of plant secondary metabolites. We discuss the role of easy bleeding, considering the fact that sawfly larvae are frequently the prey of invertebrate and vertebrate predators. It is suggested that invertebrates such as ants were more important than vertebrates in the evolution of easy bleeding. PMID- 12647189 TI - Avian fruit preferences across a Puerto Rican forested landscape: pattern consistency and implications for seed removal. AB - Avian fruit consumption may ensure plant reproductive success when frugivores show consistent preference patterns and effectively remove and disperse seeds. In this study we examined avian fruit preferences and their seed-removal services at five study sites in north-central Puerto Rico. At each site, we documented the diet of seven common fruit-eating avian species from February to September 1998. Using foraging observations and area-based estimates of fruit abundance, we examined preference patterns of birds. We found that 7 out of 68 fleshy-fruited plant species were responsible for most of the fruit diet of birds. Seventeen plant species were preferred and four of them were repeatedly preferred across several study sites and times by at least one avian species. Preferred plant species comprised a small percentage of fleshy fruits at each site (<15% in four out of five study sites), but showed extended phenology patterns. The quantity of seeds removed by frugivore species was not strictly related to preferences. Some frugivores showing no preference could effectively remove more seeds from plants at some locations than species exhibiting constancy in their patterns of preference. Only two frugivores, Euphonia musica and Vireo altiloquous, removed most of the seeds of plants for which they exhibited repeated preference across the landscape. Preference patterns, particularly those exhibiting consistency in space and time for plant species having prolonged fruiting periods, may have important mechanistic consequences for the persistence, succession, and regeneration of tropical plant communities. PMID- 12647190 TI - Controls of fine root dynamics across a gradient of gap sizes in a pine woodland. AB - Controls of fine dynamics were investigated in a Pinus palustris Mill. (longleaf pine) woodland subjected to two understory vegetation treatments (control versus removed) and four overstory treatments (no gap control, and canopy gaps of three sizes with constant total gap area per stand). Fine root (<2 mm diameter) dynamics were measured over 11 months using ingrowth cores (all treatments) and minirhizotrons (understory removed in no gap control and large gap treatments only). At the fine (microsite) spatial scale, pine and non-pine root mass production responded negatively to each other (P=0.033). Each life form was significantly (P< or =0.028) related to nearby overstory density, and pine root production compensated for reductions in non-pine roots if understory vegetation was removed. Soil moisture and NO(3) mineralization rate were negatively related to pine root mass production (ingrowth cores; P<0.001 and P=0.052) and positively related to pine root length production, mortality and turnover (minirhizotrons; P from <0.001 to 0.078). Temperature variance was negatively related to pine root lifespan P<0.001) and positively related to pine root turnover (P=0.003). At the ecosystem scale, pattern of overstory disturbance (gap size and number) had no significant effect on non-pine, pine, or total root production. However, the presence of gaps (versus the no-gap control) increased non-pine root mass production (ANOVA, P=0.055) in natural understory conditions, and reduced pine root mass production (P=0.035) where the understory was removed. Ecosystem-wide pine root length production, mortality and turnover were positively related to weekly soil temperature (P< or =0.02). In natural systems, fine root dynamics are highly variable and strongly affected by biotic factors. Roots quickly close belowground gaps because one life form (pine or non-pine) compensates for the absence of the other. When understory vegetation is removed, however, pine roots respond to the local abiotic environment, particularly moisture and NO(3). PMID- 12647191 TI - Intraguild interference and biocontrol effects of generalist predators in a winter wheat field. AB - Arable land typically harbours communities of polyphagous invertebrate natural enemies, among them numerous soil-surface dwelling predators such as ground beetles (Carabidae) and spiders (Lycosidae, Linyphiidae). Numbers of these predators were experimentally manipulated in a winter wheat field in order to study the predation impact of a generalist predator assemblage on herbivorous insects, the possible interferences among the predators concerned, and subsequent effects on wheat plant parameters. Removing ground beetles doubled numbers of Lycosidae indicative of intraguild interference between these two predator groups. Aphid densities were highest in carabid removal plots implying a substantial predation impact of ground beetles on the pest population. The predation impact of ground beetles was strongest earlier and disappeared later in the season. In mid-season, at intermediate aphid densities, the combined impact of carabid beetles and spiders appeared to be responsible for the reduction in aphid abundance. This result was probably due to a biomass effect rather than to a synergistic effect of the predator community. Thysanoptera decreased when spiders were removed (perhaps because spiders were preying on a predator of thrips), while Cicadellidae and Delphacidae showed no effect at all. The rise of aphid numbers in carabid removal plots corresponded to an increase in protein content of the wheat grains, while other plant parameters such as plant numbers and grain mass were not affected. In conclusion, this study provided field evidence for intraguild interference among generalist ground predators in arable land. Despite this interference the polyphagous predator community was able to depress numbers of aphids in winter wheat, a result cascading down to plant quality parameters. PMID- 12647192 TI - Support for the allotonic frequency hypothesis in an insectivorous bat community. AB - The allotonic frequency hypothesis proposes that certain insectivorous bat species can prey upon moths that can hear bat echolocation calls by using echolocation frequencies outside the sensitivity range of moth ears. The hypothesis predicts that the peak frequencies of bat echolocation calls are correlated with the incidence of moths in the diets of these bats. The aim of this study was to test this prediction on a bat community dominated by bats using low duty cycle echolocation calls, i.e. aerial foraging, insectivorous species using frequency modulated calls. The community consisted of nine species, two molossids, Sauromys petrophillus and Tadarida aegyptiaca, five vespertilionids, Eptesicus capensis, Eptesicus hottentotus, Miniopteris schreibersii, Myotis tricolor, and Myotis lesueuri, one rhinolophid, Rhinolophus clivosus, and one nycterid, Nycteris thebaica. The insect fauna in the habitat used by the bat community was suited to the testing of the allotonic frequency hypothesis because more than 90% of the moths comprising the insect fauna were tympanate. These included Pyralidae (3.8%), Geometridae (44.9%), Notodontidae (3.8%), Arctiidae (4.6%), Lymantriidae (0.8%) and Noctuidae (32.4%). As predicted, peak echolocation frequency was correlated with the incidence of moths in the diets of these nine species (r=0.98, df=7, P<0.01). Furthermore, multivariate analysis revealed that echolocation frequency (t=9.91, n=129, P<0.001) was a better predictor of diet than forearm length (t=5.51, n=129, P<0.001) or wing area (t= 3.41, n=129, P<0.001). This suggests that the selection pressure exerted by moth hearing might have acted directly on call frequency and secondarily on body size and wing morphology, as part of the same adaptive complex. It is unlikely that dietary differences were due to temporal and spatial differences in the availability of prey because the pattern of differences in skull morphology of the nine species supported our dietary analyses. The skull morphology of a bat represents a historical record of the kind of diet it has become adapted to over its evolutionary history. These results suggest that prey defences may mediate other factors structuring bat communities, e.g. competition. Competition may be reduced for those species of bats that can circumvent prey defences. PMID- 12647193 TI - Early use of high frequency ventilation in the premature neonate. AB - This study evaluated whether the early use of high frequency ventilation (HFV) decreased the incidence of oxygen dependency at 36 weeks postconceptual age [chronic lung disease (CLD)] and improved developmental outcome. Neonates of less than 32 weeks gestational age needing ventilatory support for RDS who were admitted to a tertiary academic neonatal intensive care unit (NICU) within 6 h of birth were included in a prospective controlled clinical trial. With randomisation they were given either HFV (n=147) or conventional ventilation (CV) (n=153). As a primary outcome variable, ventilator and/or oxygen dependence at a postconceptual age of 36 weeks (CLD) was measured. Secondary outcome variables were: mortality at discharge, treatment failure, ventilator and/or oxygen dependence at 28-30 days (bronchopulmonary disease [BPD]), duration of ventilation, use of surfactant, days in oxygen and on continuous positive airway pressure (CPAP), survival without BPD or CLD, air leak, intracranial haemorrhages (ICH) grades 3 and 4, periventricular leukomalacia (PVL) grades 1 and 2, retinopathy of prematurity (ROP), patent ductus arteriosus (PDA), necrotising enterocolitis (NEC), developmental outcome at 7 to 12 months and if necessary at 18-24 months corrected age. The results showed that CLD (16.3 vs. 12.4%), BPD (33.3 vs. 36.6%), early cerebral abnormalities, mortality at discharge (17.2 vs. 13.2%), failure rate (11.6 vs. 6.5%) and motor and mental developmental outcome at a corrected age of 18 to 24 months (p>0.05) did not differ between the two groups. CONCLUSION: Under the present study design HFV compared with CV did not decrease chronic lung disease and no developmental outcome differences could be found at a corrected age of almost 2 years. PMID- 12647194 TI - Minimal handling and bronchopulmonary dysplasia in extremely low-birth-weight infants. AB - Over the last 16 years a minitouch regime, i.e., nasal continuous positive airway pressure (n-CPAP) and/or nasal intermittent positive pressure ventilation (n IPPV), together with a minimal intubation policy has been routinely used for the treatment of respiratory distress syndrome (RDS) in preterm infants. Only 1.39 (1 out of 72) of the extremely low-birth-weight babies admitted to our Neonatal Intensive Care Unit (NICU) and surviving for at least 36 weeks' postconceptional age developed bronchopulmonary dysplasia at 36 weeks (BPD 36-wk). The BPD-36 wk incidence observed in our population is significantly lower than expected (30%) from the literature (p=0.000002). CONCLUSION: Our experience supports the effectiveness of the minitouch regime as a way to ventilate premature babies, reducing BPD risk. PMID- 12647195 TI - Incidence and risk factors of respiratory syncytial virus-related hospitalizations in premature infants in Germany. AB - Premature infants have an increased risk of developing complicated respiratory syncytial virus (RSV) infections. Epidemiological data on RSV-related hospitalizations are a prerequisite to develop guidelines for the use of preventive measures. The objective of this study was to determine incidence and risk factors of RSV-related rehospitalizations (RSV-RH) of premature infants. We recruited 1,103 infants with a gestational age of less than 35 weeks, primarily admitted to nine neonatologic care units in southern Germany between Nov. 1, 1998 and Oct. 31, 1999. Questionnaires were sent to all parents of infants discharged from neonatal care units to determine the risk of rehospitalization for acute respiratory infections (ARI-RH) and RSV-RH in the 1999-2000 season. The questionnaire response rate was 68.4%. The 717 included infants of the responders had a mean gestational age of 31.6 weeks (Range: 23-35) and a mean birth weight of 1,747 g (range: 430-4,050 g). The risk for an ARI-RH was 10.6% and the risk for RSV-RH 5.2% during the observation period. Premature infants with chronic lung disease (CLD) had a probability of 24.5% for ARI-RH and of 15% for RSV-RH. The following factors were independently associated with an increased risk of RSV RH: male gender (adjusted Odds-Ratio (OR): 8.7; 95% confidence interval (CI): 2.6 29.1), chronic lung disease (OR: 3.99; 95%CI: 1.4-11.2), discharge between October and December (OR: 2.1; 95%CI: 0.99-4.4), day-care attendance of siblings (OR: 3.9; 95%CI: 1.9-8.3). CONCLUSIONS: The risk for RSV rehospitalization among premature infants discharged from neonatal care facilities in southern Germany was low. Additional risk factors and high costs of prophylaxis have to be considered when infants are selected for RSV prophylaxis using monoclonal antibodies. PMID- 12647196 TI - Economic evaluation of possible prevention of RSV-related hospitalizations in premature infants in Germany. AB - Palivizumab (Synagis, MedImmune Inc./Abbott Laboratories) has been shown to reduce the number of respiratory syncytial virus (RSV)-related hospitalizations in premature infants. The cost-effectiveness ratio of this prophylaxis, however, has not been evaluated in the German health-care system to date. The aim of the study was to assess the costs and benefits of Palivizumab among premature infants 6) manifested longer survival than patients with lower PTHrP expression (IRS< or =6; Cox'es F test, P<0.05). Moreover, in the group with the lower PTHrP expression, a negative relationship was detected between expression of the protein and the survival time (Cox'es model, P<0.05). No relationship was detected between PTHrP expression and the development of distant metastases. PMID- 12647215 TI - Early pathogenesis of cardiac amyloid deposition in senile systemic amyloidosis: close relationship between amyloid deposits and the basement membranes of myocardial cells. AB - Despite a number of in vitro studies of transthyretin (TTR) amyloidogenesis the early stage of in vivo amyloidogenesis in the human heart is largely unknown. A heart with a mild degree of cardiac amyloidosis removed from a 90-year old woman at autopsy was selected for analysis. The genotype of the TTR was the wild type. An immunohistochemical study with anti-TTR antibody was performed on serial paraffin sections, and 17 TTR-positive lesions less than 50 micro m in diameter consisting of 13 interstitial and 4 vascular lesions were identified. The early interstitial lesions start as thick membranous deposits between interfacing myocardial cells. They are Congophilic with green birefringence and positive for apolipoprotein E but negative for amyloid P component. The TTR-positive amyloid extends along intercellular spaces and becomes larger, involving several myocardial fibers. The media is the initial site of arteriolar involvement. According to the in vitro studies of amyloid fibrillogenesis, the most critical step is formation of the nucleus under supersaturated conditions. The supersaturated conditions are speculated to be achieved by binding to proteoglycans or lipid membranes. Our results indicate that the basement membrane of myocardial cells is the initial site of amyloid deposition, providing a suitable place for concentration of TTR. PMID- 12647216 TI - Mixed ductal-endocrine carcinomas of the pancreas and ductal adenocarcinomas with scattered endocrine cells: characterization of the endocrine cells. AB - We compared the histological and immunohistochemical features of mixed ductal endocrine carcinomas of the pancreas with those of ductal adenocarcinomas (DACs) containing scattered tumor-associated endocrine cells (SECs). Three pancreatic neoplasms fulfilled the WHO criteria for mixed ductal-endocrine carcinomas. Two of them showed moderately to poorly differentiated glandular structures composed of both mucin producing and neuroendocrine cells. The third mixed ductal endocrine carcinoma was of the composite type showing DAC structures and a solid component with small epithelial cells, most of them of neuroendocrine nature. In 32 of 34 cases of DAC located in the head (30 cases) and body to tail (4 cases) of the pancreas and showing lymph-node metastases, SECs were found, but they were few in number and irregularly distributed in the tumors. In three DACs a few SECs were also detected in lymph-node metastases. Double staining for chromogranin A and the proliferation marker Ki-S5 revealed that all SECs that were not intimately integrated into the neoplastic glandular epithelium failed to show proliferative activity and changes of the expression of tumor suppressor genes (p53 and DPC 4). These findings suggest that only those SECs that belong to the proliferative cell fraction may be of neoplastic origin, while the majority of SECs probably constitute a tumor-associated but non-neoplastic cell population. These features contrast with those of mixed ductal-endocrine carcinomas, in which all endocrine cells are a component of the neoplasm. PMID- 12647217 TI - Expressions of two adenomatous polyposis coli and E-cadherin proteins on human colorectal cancers. AB - Mutations in the adenomatous polyposis coli (APC) gene contribute to the progression of colorectal tumorigenesis. Despite the importance, few studies regarding the localization of this protein on surgically resected human colorectal cancer specimens using immunohistochemistry have been reported so far because of the unavailability of the antibodies for this use. The goal of this study has been to provide the APC protein expression and to validate the APC molecular studies. We took advantage of an immunohistochemistry procedure of applying the unique detergent-mediated antigen retrieval technique to frozen sections and examined the expressions of one amino (N)-terminal (AC4) and one carboxy (C)-terminal APC antibody (HG2). Further, we compared the stainings of APC antibodies with those of the E-cadherin antibody using a quantitative image analysis. E-cadherin is a critical morphogenetic regulator during embryogenesis and recent evidence strongly suggests that downregulation of E-cadherin expression in cancers is associated with a high rate of invasion and metastasis. The analysis indicated statistically that normal epithelia showed stronger staining than cancer cells ( P<0.05). Further, in normal epithelia, the amino (N) terminal APC antibody (AC4) showed a positive correlation with another carboxy (C)-terminal APC antibody (HG2). E-cadherin showed no positive correlation with other APCs in either the normal epithelia or cancer cells. This study verified reduced expressions of APCs and E-cadherin proteins in colorectal cancer cells. This suggests that the normal APC and E-cadherin protein expressions in benign epithelium are progressively and independently lost in the sporadic colorectal cancers. PMID- 12647218 TI - Immunoprofile of cervical and endometrial adenocarcinomas using a tissue microarray. AB - Adenocarcinomas of the uterine cervix show a wide range of morphological features, and can be confused with endometrial adenocarcinoma in biopsy or curetting specimens. The objective of this study was to use tissue microarray technology to evaluate the immunoprofile of a large set of uterine adenocarcinomas with an extended panel of antibodies, comparing the profile of primary cervical and endometrial adenocarcinomas. A tissue microarray was constructed using paraffin-embedded, formalin-fixed tissues from 141 hysterectomy specimens. Duplicate 0.6-mm cores were obtained from 57 cervical adenocarcinomas (16 in situ and 41 invasive) and 84 endometrial adenocarcinomas. Tissue array sections were immunostained with 21 commercially available antibodies [B72.3, CD 99, carcinoembryonic antigen (CEA), c-kit, pancytokeratin, CK 5/6, CK 7, CK8/18, CK19, CK 20, CK 22, EMA, estrogen receptor (ER), KP-1, melan-A, p53, PLAP, S-100, synaptophysin, TTF-1, and vimentin] utilizing the avidin-biotin (ABC) technique. Hierarchical clustering analysis of the tumors was done based on the immunostaining results. Only ER ( P<0.001), CEA ( P=0.04), vimentin ( P<0.001), and CK 8/18 ( P=0.002) showed a significantly different frequency of positivity in endometrial relative to cervical adenocarcinomas. ER, vimentin, and CK 8/18 were more likely to be expressed in endometrial adenocarcinomas, while cervical adenocarcinomas more frequently expressed CEA. We were able to identify immunoprofiles that were highly specific for endocervical adenocarcinoma (ER(-), vimentin(-), CK 8/18(-), CEA(+)) or endometrial adenocarcinoma (ER(+), vimentin(+), CK 8/18(+), CEA(-)), but most tumors showed an intermediate, non specific immunophenotype. Hierarchical clustering analysis was useful in the interpretation of these intermediate immunophenotypes. Papillary serous adenocarcinoma of the endometrium was less likely to express vimentin ( P=0.002) than endometrioid carcinoma of the endometrium. PMID- 12647219 TI - Expression of promyelocytic leukemia protein increases during the differentiation of human neuroblastoma cells. AB - The promyelocytic leukemia (PML) protein, whose fusion with retinoic acid receptor alpha is responsible for the tumorigenesis of acute promyelocytic leukemia, acts as a tumor suppressor in various types of human cancers. We analyzed the expression patterns of PML, in both primary neuroblastic tumors ( n=20) and two human neuroblastoma (NB) cell lines, SMS-KCNR (KCNR) and SH-SY5Y (SY5Y). The expression of PML, revealed as speckled or microgranular staining in the nuclei, was positively correlated with the differentiation status of NB cells in vivo, and was upregulated during the differentiation of KCNR and SY5Y cells following retinoic acid treatment. Screening of PML expression in human brain and sympathetic ganglia showed restricted expression of PML in mature neurons and glial cells, a result that was consistent with that in differentiated NB tumors. All these findings strongly suggest that increased PML expression is associated with growth inhibition and differentiation of human NB cells, and that it is of critical significance in the biology of NBs and in human nervous system development. PMID- 12647221 TI - Multiparameter flow cytometry in the diagnosis of a gynaecologic double tumor: a case report. AB - PURPOSE: An uncommon clinical presentation of metastatic tumor will often lead to additional diagnostic examinations. The patient of the present study was known to have endometrial cancer which was thought to be limited to the endometrium. Three months postoperatively, she developed ascites due to spread of the tumor, which is rarely seen in low-stage endometrial cancer. METHOD: Multiparameter flow cytometry using both cell phenotype information and DNA ploidy was performed. RESULTS: Retrospectively, the patient was diagnosed as having a DNA-diploid epithelial tumor of the endometrium as well as a DNA-aneuploid epithelial tumor in the left fallopian tube. It was shown that 3 months after primary surgery she developed ascites caused by metastatic tumor from the primary fallopian tube cancer. CONCLUSION: The complete diagnosis was made using multiparameter flow cytometry which, at present, is not routinely applied in gynecologic pathology. PMID- 12647220 TI - Expression patterns of transcription factors in progressively transformed germinal centers and Hodgkin lymphoma. AB - The World Health Organization (WHO) classification of Hodgkin lymphoma (HL) distinguishes two types: Classical Hodgkin lymphoma (CHL) and nodular lymphocyte predominant Hodgkin lymphoma (NLPHL). Both groups have in common that they mostly derive from B cells with rare classical cases originating from T cells. They differ in their histomorphology, immunophenotype, and clinical behavior. One of the subtypes of CHL, designated as lymphocyte-rich classical Hodgkin lymphoma (LRCHL), shares some morphological features with NLPHL. The transcription factors BSAP, BOB.1, Oct2 and MUM1 are sequentially expressed in normal B-cell development. In order to investigate the relationship between the CHL subgroups and NLPHL, we examined the protein expression of these transcription factors using immunohistochemistry in 15 reactive processes and 4 different subtypes of 58 HL cases. Our findings underline the B-cell origin of HL, without evidence, that reactive processes like progressively transformed germinal centers (PTGCs) are precursor lesions of HL. Furthermore, they demonstrate that LRCHL is distinct from NLPHL and that it is closely related to the mixed cellularity CHL (MCHL) in respect of BSAP, BOB.1, and Oct2 expression. It therefore occupies an intermediate position between MCHL and NLPHL. Based on MUM1 staining, LRCHL exhibits a more mature phenotype than NLPHL. PMID- 12647222 TI - Cutaneous papillary squamous cell carcinoma. Report of three new cases and review of the literature. AB - Squamous cell carcinoma is one of the most common malignant cutaneous tumors. Several histological variants have been described; the papillary subtype is one of the most infrequent, with only four cases having being reported previously. We report three new cases of this unusual variant of cutaneous squamous cell carcinoma, review the literature and consider the main differential diagnoses. Polymerase chain reaction performed in our three cases did not demonstrate human papilloma virus infection. PMID- 12647223 TI - Ectomesenchymal chondromyxoid tumor of the anterior tongue with myxoglobulosislike change. PMID- 12647224 TI - Roles of dynamic linkage of stable attractors across cortical networks in recalling long-term memory. AB - We propose a neural network model for a category-association task. By simulating the model, neuronal relevance of cortical interactions to recalling long-term memory was investigated. The model consists of the left and right hemispheres, each of which has IT (inferotemporal cortex) and PC (prefrontal cortex) networks. Information about visual features and their categories were encoded into point attractors of the IT and PC networks, respectively. In the task, the IT network of the right hemisphere was stimulated with a cue feature. After a delay period, the IT network of the left hemisphere was simultaneously stimulated with the choice feature and an irrelevant feature. The cue and choice features belong to the same category, while the irrelevant feature belongs to another category. To complete the task, the IT network must select the point attractor corresponding to the choice feature. We demonstrate that the top-down pathway (PC-to-IT) triggers the retrieval of long-term memory of the choice feature from the IT, and the bottom-up pathway (IT-to-PC) contributes to the maintenance of the retrieved memory during the delay period. The key mechanism for the retrieval and maintenance of that memory is the dynamic linkage of attractors across separate cortical networks. We show that a single hemisphere is sufficient for the memory retrieval, but it is advantageous to use the two hemispheres because the retrieved memory is thereby retained with greater reliability until the brain chooses the choice feature. PMID- 12647225 TI - Use of 'relative-phase' analysis to assess correlation between neuronal spike trains. AB - We propose a new method of studying the correlation between neuronal spike trains. This technique is based on the analysis of relative phase between two point processes. Relative phase here is defined as the relative timing difference between two spike trains normalized by the associated interspike interval of one cell. This phase measurement is intended to reveal the relative timing relationship between spike trains at different firing rates. We apply this method to a numerical example and an example from two cerebellar neuronal spike trains of a behaving rat. The results are compared with classical cross-correlation analysis. We show that the technique can avoid some of the limitations of cross correlation methods, reveal certain statistical dependencies that cannot be shown by cross correlation, and provide information as to the direction of influence between two spike trains. PMID- 12647226 TI - A space-variant filter model of texture segregation: parameter adjustment guided by psychophysical data. AB - This article presents a space-variant version of a standard spatial filter model of texture segregation of the "back-pocket" type (i.e., two filter layers with an intermediate pointwise nonlinearity). The model was tested with psychophysical data from experiments with line textures in which target lines differed in orientation from background lines. The textures were presented briefly and then masked. Segregation performance was evaluated along the horizontal meridian up to retinal eccentricities of about 10 deg. Data are reported from two experiments with different line densities (Kehrer 1989) and two experiments with different orientation contrasts between target lines and background lines (Kehrer 1990). Segregation performance proved to depend strongly on these texture variations, and it peaked several degrees from fixation in all cases. The filter model provided satisfactory predictions of experimental data when model parameters were adjusted appropriately. It is concluded (1) that filter models defined in strictly spatial terms (i.e., without temporal properties) offer a sufficient framework to account for the psychophysical data and (2) that the particular course of the performance curve (i.e., the performance peak outside the central region) must be attributed to the characteristics of second-layer filters. PMID- 12647227 TI - Synaptic fusion pore structure and AMPA receptor activation according to Brownian simulation of glutamate diffusion. AB - The rising phase of fast, AMPA-mediated Excitatory Post Synaptic Currents (EPSCs) has a primary role in the computational ability of neurons. The structure and radial expansion velocity of the fusion pore between the vesicle and the presynaptic membrane could be important factors in determining the time course of the EPSC. We have used a Brownian simulation model for glutamate neurotransmitter diffusion to test two hypotheses on the fusion pore structure, namely, the proteinaceous pore and the purely lipidic pore. Three more hypotheses on the radial expansion velocity were also tested. The rising phases of the EPSC, computed under various conditions, were compared with experimental data from the literature. Our present results show that a proteinaceous fusion pore should produce a more marked foot at the beginning of the rising phase of the EPSC. They also confirm the hypothesis that the structure of the fusion pore and its radial expansion velocity play significant roles in shaping the fast EPSC time course. PMID- 12647228 TI - Deriving physical connectivity from neuronal morphology. AB - A model is presented that allows prediction of the probability for the formation of appositions between the axons and dendrites of any two neurons based only on their morphological statistics and relative separation. Statistics of axonal and dendritic morphologies of single neurons are obtained from 3D reconstructions of biocytin-filled cells, and a statistical representation of the same cell type is obtained by averaging across neurons according to the model. A simple mathematical formulation is applied to the axonal and dendritic statistical representations to yield the probability for close appositions. The model is validated by a mathematical proof and by comparison of predicted appositions made by layer 5 pyramidal neurons in the rat somatosensory cortex with real anatomical data. The model could be useful for studying microcircuit connectivity and for designing artificial neural networks. PMID- 12647229 TI - Robustness of neuroprosthetic decoding algorithms. AB - We assessed the ability of two algorithms to predict hand kinematics from neural activity as a function of the amount of data used to determine the algorithm parameters. Using chronically implanted intracortical arrays, single- and multineuron discharge was recorded during trained step tracking and slow continuous tracking tasks in macaque monkeys. The effect of increasing the amount of data used to build a neural decoding model on the ability of that model to predict hand kinematics accurately was examined. We evaluated how well a maximum likelihood model classified discrete reaching directions and how well a linear filter model reconstructed continuous hand positions over time within and across days. For each of these two models we asked two questions: (1) How does classification performance change as the amount of data the model is built upon increases? (2) How does varying the time interval between the data used to build the model and the data used to test the model affect reconstruction? Less than 1 min of data for the discrete task (8 to 13 neurons) and less than 3 min (8 to 18 neurons) for the continuous task were required to build optimal models. Optimal performance was defined by a cost function we derived that reflects both the ability of the model to predict kinematics accurately and the cost of taking more time to build such models. For both the maximum-likelihood classifier and the linear filter model, increasing the duration between the time of building and testing the model within a day did not cause any significant trend of degradation or improvement in performance. Linear filters built on one day and tested on neural data on a subsequent day generated error-measure distributions that were not significantly different from those generated when the linear filters were tested on neural data from the initial day (p<0.05, Kolmogorov-Smirnov test). These data show that only a small amount of data from a limited number of cortical neurons appears to be necessary to construct robust models to predict kinematic parameters for the subsequent hours. Motor-control signals derived from neurons in motor cortex can be reliably acquired for use in neural prosthetic devices. Adequate decoding models can be built rapidly from small numbers of cells and maintained with daily calibration sessions. PMID- 12647230 TI - Dynamical synaptic plasticity: a model and connection to some experiments. AB - Using a modified version of a phenomenological model for the dynamics of synaptic plasticity, we examine some recent experiments of Wu et al. [(2001) J Physiol 533:745-755]. We show that the model is quantitatively consistent with their experimental protocols producing long-term potentiation (LTP) and long-term depression (LTD) in slice preparations of rat hippocampus. We also predict the outcome of similar experiments using different frequencies and depolarization levels than reported in their results. PMID- 12647231 TI - Suppression of contour perception by band-limited noise and its relation to nonclassical receptive field inhibition. AB - Band-spectrum noise has been shown to suppress the visual perception of printed letters. The suppression exhibits a specific dependence on the spatial frequency of the noise, and the frequency domain of most effective inhibition has been related to the size of the letters. In this paper, we address two important questions that were left open by previous studies: (1) Is the observed effect specific to text, and which parameters determine the domain of most effective suppression? (2) What is the origin of the effect in terms of underlying neural processes? We conduct a series of psychophysical experiments that demonstrate that the frequency domain of most effective inhibition depends on the stroke width of the letter rather than on the letter size. These experiments also demonstrate that the effect is not specific to the recognition of letters but also applies to other objects and even to single bars. We attribute the observed effect to nonclassical receptive field (non-CRF) inhibition in visual area V1. This mechanism has previously been suggested as the possible origin of various other perceptual effects. We introduce computational models of two types of cell that incorporate non-CRF inhibition, which are based on Gabor energy filters extended by surround suppression of two kinds: isotropic and anisotropic. The computational models confirm previous qualitative explanations of perceptual effects, such as orientation contrast pop-out, reduced saliency of lines embedded in gratings, and reduced saliency of contours surrounded by textures. We apply the computational models to the images used in the psychophysical experiments. The computational results show a dependence of the inhibition effect on the spatial frequency of the noise that is similar to the suppression effect measured in the psychophysical experiments. The experimental results and their explanation give further support to the idea of a possible functional role of non-CRF inhibition in the separation of contour from texture information and the mediation of object contours to higher cortical areas. PMID- 12647233 TI - Inhaled ultrasonically nebulized distilled water decreases exhaled nitric oxide in asthma. AB - Exhaled nitric oxide (eNO) is increasingly used as a marker of disease activity in asthma. Inhaled hypertonic saline has been shown to induce bronchoconstriction and to decrease eNO in asthmatic subjects, whereas the effects of hypotonic solutions on eNO in these patients have not been studied. To evaluate the effect of ultrasonically nebulized distilled water (UNDW), an indirect hypotonic stimulus, on eNO, 17 asthmatic patients were enrolled and eNO from lower airways was measured by chemiluminescence. UNDW significantly reduced FEV(1) >or= 20% in 9 subjects (UNDW+), but had no effect in eight patients (UNDW-). Baseline eNO concentration were found to be 51.3 +/- 11.1 ppb in UNDW+ and 32.9 +/- 7.5 ppb in UNDW- patients, respectively ( p = 0.199, NS). UNDW inhalation significantly decreased eNO (from 51.3 +/- 11.1 ppb to 31.0 +/- 7.1 ppb in UNDW+ ( p < 0.020, n = 9) and from 32.9 +/- 7.5 ppb to 26.2 +/- 7.3 ppb in UNDW- subjects ( p < 0.024, n = 8), respectively). eNO percentage reduction in UNDW+ patients was significantly higher compared with UNDW- subjects (-37 +/- 4% vs -23 +/- 3%, p = 0.021). There was no correlation between FEV(1) changes and eNO percentage decreases in both UNDW+ and UNDW- subjects. In UNDW+ patients, acute bronchodilation induced by salbutamol caused a recovery in both FEV(1) and eNO, though eNO levels remained lower than baseline values. We concluded that UNDW inhalation can significantly decrease eNO in asthmatic patients, either responders or nonresponders to this indirect osmotic challenge; the reduction in eNO levels was only partly dependent on acute changes in airway caliber. PMID- 12647234 TI - Mannitol dose-dependently attenuates lung reperfusion injury following liver ischemia reperfusion: a dose-response study in an isolated perfused double-organ model. AB - We had previously studied different modes of prevention of liver ischemia reperfusion (IR)-induced remote organ reperfusion injury, a challenge that remains partly unmet. We have now studied the capability of mannitol at different doses in abrogating liver IR-induced lung reperfusion injury in an isolated double-organ model. Rat livers ( n = 8/group) were perfused with Krebs-Henseleit solution (control) or made globally ischemic (IR) for 2 h, after which they were paired with normal lungs and "reperfused" together for 15 min. The lungs were then perfused alone with the accumulated Krebs for an additional 45 min. Another 4 control and 4 IR pairs were reperfused with Krebs containing mannitol at.22 mmol,.55 mmol,.77 mmol, or 1.1 mmol. Mannitol.22 mmol and 1.1 mmol failed to attenuate IR-lung injury as indicated by 50-95% increases in inspiratory and perfusion pressures and compliance reduction, a 70% increase in weight gain, and a 2-50-fold increase in bronchoalveolar lavage volume and content. Mannitol.55 mmol prevented all these abnormalities, and.77 mmol attenuated only changes in ventilatory parameters. The latter two treatments were also associated with a 50% reduction in xanthine oxidase activity and a 35-45% increase in the reduced glutathione tissue content compared with the nontreated IR-paired lungs. It is concluded that mannitol in a narrow therapeutic dose range can reduce oxidalive stress-induced lung damage that is related to liver IR. PMID- 12647232 TI - Hyperoxia and nitric oxide reduce surfactant components (DSPC and surfactant proteins) and increase apoptosis in adult and fetal rat type II pneumocytes. AB - Nitric oxide (NO) alone or in conjunction with hyperoxia can have protective or detrimental effects on the lung. Our hypothesis was that hyperoxia in conjunction with NO would result in increased cellular dysfunction and apoptotic cell death in adult and fetal Type II pneumocytes (TIIP) in a dose-dependent manner. The TIIP were obtained from adult and 19-day fetal rat lungs. The TIIP were then exposed to 100, 200 and 500 micro M of the NO-donor, Glyco-SNAP-2, alone or in conjunction with 95% oxygen for 24 h. While low-dose NO exposure alone did not increase cytotoxicity, in conjunction with hyperoxia, there was a significant dose-dependent increase in apoptotic cell death of adult TIIP as well as fetal TIIP. Choline incorporation into disaturated phosphatidylcholine was markedly decreased in adult TIIP while the fetal TIIP had similar values as controls. However, the mRNAs of surfactant proteins A, B and C as well as iNOS were significantly reduced in fetal TIIP. Exogenous peroxynitrite also increased nitrotyrosine formation in fetal TIIP as did hyperoxia and NO. The effect of hyperoxia and NO could be abrogated with catalase and superoxide dismutase. These findings may have significant clinical implications in the use of NO in premature infants. PMID- 12647235 TI - Antileukinate, a hexapeptide inhibitor of CXC-chemokine receptor, suppresses bleomycin-induced acute lung injury in mice. AB - A hexapeptide, Ac-RRWWCR-NH(2) (Antileukinate), has been reported to be a potent inhibitor of CXC-chemokine receptor. However, the in vivo anti-inflammatory activity of this agent has not been tested except in a rabbit skin edema model. This study was undertaken to investigate the effect of subcutaneously administered Antileukinate on experimental bleomycin-induced acute lung injury in mice, in which CXC-chemokines have been reported to be involved. Lung injury was assessed on the basis of histopathology, the number of total cells, the percentage of neutrophils, and protein concentration in the bronchoalveolar lavage (BAL) fluid, and the wet lung weight at 7 days after intratracheal instillation of bleomycin. Histopathological studies revealed that treatment with Antileukinate markedly suppressed inflammatory cell infiltration and interstitial lung edema. The neutrophil counts in the BAL fluid were significantly decreased in the Antileukinate-treated group. The suppression of pulmonary edema was further confirmed by the reduction of wet lung weight and total protein concentration in the BAL fluid. These findings suggest that Antileukinate is able to inhibit acute lung injury by suppressing neutrophil mobilization induced by CXC-chemokines. PMID- 12647236 TI - Chest wall kinematics during chemically stimulated breathing in healthy man. AB - Chest wall compartment kinematics and respiratory muscle coordinate activity, during either hypercapnia or hypoxia, have not been comparatively assessed in healthy humans. We assessed the displacement volume of the chest wall (Vcw) in 5 normal subjects during hypoxic-normocapnic and hypercapnic-hyperoxic rebreathing by using linearized magnetometers. Vcw was divided into displacement volumes of the rib cage (Vrc) and the abdomen (Vab). Esophageal (Pes) and gastric (Pga) pressures were simultaneously recorded and transdiaphragmatic pressure (Pdi) was calculated by subtracting Pes from Pga. Pressure swings (sw) from end expiration (EE) to end inspiration (EI) were also calculated. During both hypoxia and hypercapnia, from quiet breathing to 40 L/min VE, Vrc,EI increased consistently but Vrc,EE, and Vab,EI did not. Moreover, Vab,EE decreased significantly during hypercapnia and remained unchanged during hypoxia. PesEI decreased (more negative values) and PesEE increased (less negative values) during either stimulus, while PgaEE increased with hypercapnia. Pdisw, calculated as the difference between PdiEE and PdiEI, increased significantly with both hypercapnia and hypoxia ( p = 0.002 for both). On the plot of Pes vs Pga, the slope of a line from end expiratory to end inspiratory lung volume between 20 and 40 L/min VE progressively increased during hypercapnia indicating increasing rib cage muscle (RCM) contribution to inspiratory pressure swings relative to the diaphragm. From these results we conclude that in healthy man: (i) with both chemical stimuli RCM contribution accounts for increase in Vrc displacement; (ii) with hypercapnia, the decrease in Vab,EE displacement indicates abdominal muscle (ABM) contribution to tidal volume; (iii) RCM and ABM assist the diaphragmatic function during hypercapnic stimulation. PMID- 12647238 TI - Genetic and phenotypic analysis of the mouse mutant mh2J, an Ap3d allele caused by IAP element insertion. AB - Mocha (mh), a mouse model for Hermansky-Pudlak syndrome (HPS), is characterized by platelet storage pool deficiency, pigment dilution, and deafness as well as neurological abnormalities. The trans-Golgi/endosome adaptor-related complex AP-3 is missing in mh mice owing to a deletion in the gene encoding the delta subunit. Mice mutant for a second allele, mh(2J), are as hyperactive as mh, and display both spike wave absence and generalized tonic clonic seizures, but have less coat color dilution, no hearing loss, and no hypersynchronized EEG. Here we show that the mh(2J) mutation is due to an IAP element insertion in the Ap3d gene leading to a C-terminally truncated protein. Despite correct assembly of the AP-3 complex and localization to the trans-Golgi network and endosomes, AP-3 function in neurons remains impaired. While mh mice show a severe reduction of vesicular zinc (TIMM staining) owing to mislocalization and degradation of the Zinc transporter ZnT-3, the TIMM and ZnT-3 staining patterns in mh(2J) varies, with normal expression in hippocampal mossy fibers, but abnormal patterns in neocortex. These results indicate that the N-terminal portion of the delta subunit is sufficient for AP-3 complex assembly and subcellular localization to the TGN/endosomes, while subsequent function is regulated in part by cell-specific interactions with the C-terminal portion. PMID- 12647239 TI - Effects of spontaneous KitlSteel mutations on survival and red blood cells of mice. AB - Kit ligand (Kitl), which is a member of the helical cytokine superfamily, is encoded by the Steel (Sl) locus of mice and is essential for the development of hematopoietic cells, germ cells, and melanocytes. A large series of Kitl(Sl) alleles has been described, including some that arose spontaneously and others that were induced by either chemical or radiation mutagenesis. Here we describe the nucleotide sequence alterations in two spontaneous Kitl(Sl) alleles. The Kitl(Sl-18R) allele has a point mutation that introduces a premature termination codon, and the encoded protein is expected to be null functionally. The Kitl(Sl 5R) allele has an in-frame deletion that results in deletion of amino acids at position 31 and 32 of Kitl. While both mutations exert severe effects on blood cells and survival of homozygous mice, these effects are slightly milder than those of a previously characterized spontaneous deletion allele, Kitl(Sl-gb). Examination of the survival of compound heterozygotes provided strong genetic evidence that the Kitl(Sl-18R) and Kitl(Sl-5R) mutants are null functionally for mouse survival. PMID- 12647240 TI - Characterization of a QTL affecting skeletal size in mice. AB - Previous work identified a tail length QTL on Chromosome (Chr) 1 in an F(2) population of C57BL/6J x DBA/2J mice. The goals of the present study were to (1) refine the position of this QTL by additional genotyping of samples from the original study; (2) confirm the effect of this QTL by producing a partially congenic strain carrying the C57BL/6J allele against the DBA/2J background; and (3) examine the effect of the QTL on skeletal dimensions. The presence of the QTL was confirmed in a new F(2) population (N = 431) derived from the partially congenic strain, and estimates of its additive effects were similar to those from the original F(2) population (N = 901) in both sexes, i.e., the C57BL/6J chromosomal segment increased tail length, the additive effect (half the difference between homozygotes) being 0.5-0.8 standard deviations. The QTL region was more than halved, relative to that in the previous study, to an 8-cM region between D1Mit30 and D1Mit57. Among a subsample of individuals (N = 30) from the new F(2) population that were not recombinant within the QTL region, there was a significant additive effect of the QTL on the length of the humerus, femur, tibia, mandible, scapula, pelvic girdle, and a tail bone; the direction of the effect was the same as for tail length. No significant effect was found on the number of bones in the tail or on the dimensions of the ulna, skull, or first vertebra. PMID- 12647241 TI - Chromosomal regions controlling resistance to gastro-intestinal nematode infections in mice. AB - Quantitative trait loci (QTL) associated with resistance to an intestinal worm in a well-defined murine model are described. These have been identified in an F(2) population derived from resistant (SWR) and susceptible (CBA) parental mouse strains infected with the gastro-intestinal nematode parasite Heligmosomoides polygyrus. Seven QTL located on six chromosomes are described, associated with components of the complex host response and the differential regulation of parasite survival and reproduction. The combined additive effects of the five significant QTL associated with worm survival (total worm count at necropsy) account for about 60% of the difference in worm count between the parental lines. The dominance effect for these five QTL are all in the direction of resistance, supporting the heterosis for resistance established from the mean worm count for the F(2) line relative to the parental lines. It is now possible to identify the comparative chromosomal regions of these QTL in livestock and humans and to consider the possibility of future improved control strategies. These may include breeding of resistant or tolerant livestock, development of vaccines, or identification of new anthelmintic drugs. PMID- 12647242 TI - Comparative analysis of vertebrate Shh genes identifies novel conserved non coding sequence. AB - The puffer fish Takifugu rubripes (Fugu), with its compact genome, is an ideal model organism for comparative genomics. Sonic hedgehog (Shh) is a key protein in the patterning of differentiating cells during embryonic development. We have sequenced the Fugu Shh gene and compared it with the mammalian and zebrafish orthologs, identifying a number of novel conserved, non-coding sequences upstream of exon one and within the two introns. Additional conserved sequences serve to delineate activator regions and enhancers previously characterized through functional analysis. Control elements can thus be rapidly and effectively predicted by comparative methodology in its own right as well as complementing other, functional methods. This work demonstrates the value of using Fugu in comparative genomics, which has allowed identification of new putative regulatory elements, as well as corroborating enhancers identified by the more traditional deletion mapping method. PMID- 12647243 TI - Characterization and radiation hybrid mapping of expressed sequence tags from the canine brain. AB - Maps of the canine genome are now developing rapidly. Most of the markers on the current integrated canine radiation hybrid/genetic linkage/cytogenetic map are highly polymorphic microsatellite (type II) markers that are very useful for mapping disease loci. However, there is still an urgent need for the mapping of gene-based (type I) markers that are required for comparative mapping, as well as identifying candidate genes for disease loci that have been genetically mapped. We constructed an adult brain cDNA library as a resource to increase the number of gene-based markers on the canine genome map. Eighty-one percent of the 2700 sequenced expressed sequence tags (ESTs) represented unique sequences. The canine brain ESTs were compared with sequences in public databases to identify putative canine orthologs of human genes. One hundred nine of the canine ESTs were mapped on the latest canine radiation hybrid (RH) panel to determine the location of the respective canine gene. The addition of these new gene-based markers revealed three conserved segments (CS) between human and canine genomes previously detected by fluorescence in situ hybridization (FISH), but not by RH mapping. In addition, five new CS between dog and human were identified that had not been detected previously by RH mapping or FISH. This work has increased the number of gene-based markers on the canine RH map by approximately 30% and indicates the benefit to be gained by increasing the gene content of the current canine comparative map. PMID- 12647245 TI - [Micro incision cataract surgery time has arrived]. PMID- 12647244 TI - Construction and integration of radiation-hybrid and cytogenetic maps of dog Chromosome X. AB - Chromosome (chr) X is under-represented in current maps of the genome of the domestic dog ( Canis familiaris). To address this problem, we have constructed a small-insert, genomic DNA library in pBluescript from flow-sorted canine Chr X DNA. Fluorescence in situ hybridization (FISH) studies confirmed that the library was highly enriched for Chr X. Clones containing microsatellites were identified and sequenced. Database searches detected significant sequence identity between four X-derived clones and genes previously characterized in other species. Thirty seven markers derived from these clones were mapped on Chr X by FISH, and of these, 28 were mapped by using the female-derived T72 whole-genome radiation hybrid (RH) panel (Research Genetics). Four X-linked canine genes from publicly available data were also mapped. Eight RH linkage groups with LOD >4.0 were identified, and FISH data were used to locate the groups on the chromosome; four groups could be unambiguously orientated by FISH data. In each case, the FISH and RH data were mutually consistent. The data suggest strongly conserved synteny between canine and human X Chrs. The pseudoautosomal region has been further characterized, and the putative or actual locations of nine genes of clinical relevance have been suggested. PMID- 12647246 TI - [New trends in ocular drug delivery]. PMID- 12647247 TI - [Mitomycin C treatment for ocular cicatricial pemphigoid]. PMID- 12647248 TI - [Techniques for preparing postmortem human eyes to perform anterior segment intraocular surgery]. AB - We describe different methods to prepare postmortem human or animal eyes used at the Center for Research in Ocular Therapeutics and Biodevices at the Storm Eye Institute, Medical University of South Carolina, Charleston, SC, USA. These techniques have been utilized for performing different surgical procedures (phacoemulsification, extracapsular cataract extraction, etc.), and for training of surgeons in-transition. Performing these techniques in the wet-laboratory contributed to improve surgical skills to perform the critical steps of the phacoemulsification surgery. Pathological evaluation of pseudophakic postmortem human eyes using the Miyake-Apple posterior view and histology was helpful to analyze postoperative complications of cataract surgery (anterior capsule opacification and posterior capsule opacification) secondary to postoperative proliferation of lens epithelial cells into the capsular bag. Modifications in the surgical techniques and/or lens design may be helpful to reduce these postoperative complications. Implantation of various aphakic and phakic intraocular lenses in postmortem human eyes as well as animal eyes was helpful to analyze the sizing and fitting of new lens designs within the eye. PMID- 12647249 TI - [Clinical risk factors for postoperative proliferative vitreoretinopathy (PVR). A prospective study]. AB - PURPOSE: To identify clinical risk factors for development of postoperative PVR, to determine the incidence of this complication and its time of onset by a prospective multicentric study. METHODS: A multicentric and prospective study of 223 patients with rhegmatogenous retinal detachment (RD) was conducted. Logistic regression analysis was used to identify risk factors for PVR among 83 variables related to preoperative, intraoperative and postoperative characteristics. RESULTS: 22 out of 223 RD developed PVR (incidence 9.9%, confidence interval 95%: 5.9-13.9). After logistic regression analysis, four variables showed an odds ratio higher than 1.0 (RD affecting 4 quadrants, cryopexy, aphakia/pseudophakia and those RD in which an encircling band was implanted). None of these factors showed a <

> value lower than 0.05. Most of postoperative PVR (77.2%) appeared in the first month after surgery. CONCLUSIONS: This study establishes the incidence of PVR, and its time of onset, but it was not effective to identify clinical risk factors with a high level of confidence. PMID- 12647250 TI - [Choroidal melanoma in ocular melanosis]. AB - CASES: We present four cases of ocular melanosis. Choroidal melanoma was detected in all of them. Three eyes had decreased visual acuity and were enucleated because of their large, active tumours. In the fourth case the melanoma was detected in a routine examination and we were able to apply a preserving treatment with I125 brachytherapy. DISCUSSION: Melanosis oculi is often underestimated as a risk factor for uveal melanoma and glaucoma. Ophthalmic surveillance, every 6 or 12 months is important, in patients with ocular melanocytosis for early detection of high risk diseases. PMID- 12647251 TI - [Cryptococcal choroiditis in a patient with acquired immunodeficiency syndrome]. AB - PURPOSE/METHODS: We report the case of a 31 year old male with presumed cryptococcal choroiditis associated with cryptococcal meningitis and AIDS. Multiple yellowish-whitish small choroidal lesions, retinal hemorrhages and cotton-wool spots appeared in the posterior pole associated with vision loss. We also carried out differential diagnosis with other frequent diseases and a review of the treatments. RESULTS/CONCLUSIONS: Cryptococcus neoformans is an opportunistic fungus with a predilection for infecting the meninges in patients with Acquired Immunodeficiency Syndrome AIDS (10%). Ocular manifestation usually includes cranial nerve palsies or papilledema secondary to increased intracranial pressure. Cryptococcal choroiditis is less frequent (5% of patients with meningitis). PMID- 12647253 TI - [Mutation P28T in gene GK1 as the cause of a familial galactokinase deficiency]. AB - OBJECTIVE/METHOD: To alert about galactokinase deficiency (GK) as a possible cause of infantile cataracts, and even presenile cataracts in heterozygous carriers. Diagnosis by enzyme and galactitol determination would lead to the introduction of a galactose-free diet which completely prevents the damage. RESULT/CONCLUSIONS: We report on a highly consanguineous Spanish family of gypsy ethnia, with three females of different sibships affected by GK deficiency. The deficiency was due to their homozygosis for mutation P28T in gene GK1. P28T mutation in european Romani gypsies, is also present in Spanish gypsies. It is important to bear in mind that GK deficiency may be an important cause of blindness in that endogamous group. PMID- 12647252 TI - [Chronic endophthalmitis in pseudophakic patients caused by bacillus subtilis]. AB - OBJECTIVE/METHODS: We present two cases of patients with chronic endophthalmitis. The treatment included intraocular lens exchange, total capsulectomy and posterior vitrectomy through the limbus. In both cases, samples of the vitreous were taken and sent to the laboratory together with the intraocular lens and remmants of posterior capsule for culture and antibiogram. RESULTS/CONCLUSIONS: Microbiologic examination and culture were positive in two cases for Bacillus subtilis, which is not actually identified in literature as an etiologic agent for chronic endophthalmitis. PMID- 12647254 TI - [Orbital abscess secondary to sinusitis in a child]. AB - PURPOSE/METHOD: We present the case of a 10-year-old patient who was admitted as an emergency to our hospital with right ocular infection due to pansinusitis. He presented fever and malaise. Clinical, ophthalmic and radiologic examination confirmed the existence of an orbital abscess. Due to the poor evolution, surgical treatment was needed. RESULTS/CONCLUSION: Orbital cellulitis and orbital abscess are two different types of orbital infections. Being able to differentiate between then is very important for selecting the appropriate medical and surgical therapy. Medical treatment is a good solution for infections without abscesses, but surgery is usually needed for infections with an abscess. Abscess formation sometimes require surgical drainage of the orbital abscess with associated treatment of paranasal sinus. PMID- 12647255 TI - [Neurotrophic keratopathy management with cyanoacrylate tarsorraphy and autologous serum]. AB - PURPOSE/METHODS: Neurotrophic keratopathy is a degenerative corneal disease with a highly complex treatment caused by an impairment of corneal sensitivity. We report the case of a 47 year-old man with a refractory postherpetic neurotrophic keratopathy in his right eye and we discuss the treatment options. RESULTS/CONCLUSION: The combination of cyanoacrylate with temporary tarsorraphy along with the administration of topical autologous serum eyedrops and systemic tetracycline, allowed to control the progression of the disease and to regenerate the corneal surface. This unpublished therapeutic strategy might be an effective and safe alternative in the management of neurotrophic keratopathy (Arch Soc Esp Oftalmol 2003; 78: 119-122). PMID- 12647256 TI - [Tacuinum sanitatis]. PMID- 12647257 TI - [Joseph-Antoine Ferdinand Plateau (Brussels, 1801 - Gante, 1883)]. PMID- 12647258 TI - Polymorphisms at the G72/G30 gene locus, on 13q33, are associated with bipolar disorder in two independent pedigree series. AB - Linkage evidence suggests that chromosome 13 (13q32-33) contains susceptibility genes for both bipolar disorder and schizophrenia. Recently, genes called "G72" and "G30" were identified, and polymorphisms of these overlapping genes were reported to be associated with schizophrenia. We studied two series of pedigrees with bipolar disorder: the Clinical Neurogenetics (CNG) pedigrees (in which linkage to illness had been previously reported at 13q32-33), with 83 samples from 22 multiplex families, and the National Institute of Mental Health (NIMH) Genetics Initiative pedigrees, with 474 samples from 152 families. Sixteen single nucleotide polymorphisms (SNPs) were genotyped at and around the G72/G30 locus, which covered a 157-kb region encompassing the entire complementary DNA sequences of G72 and G30. We performed transmission/disequilibrium testing (TDT) and haplotype analysis, since a linkage-disequilibrium block was present at this gene locus. In the CNG and NIMH data sets, the results of global TDT of the entire haplotype set were significant and consistent (P=.0004 and P=.008, respectively). In the CNG series, the associated genotypes divided the families into those with linkage and those without linkage (partitioned by the linkage evidence). Analysis of the decay of haplotype sharing gave a location estimate that included G72/G30 in its 95% confidence interval. Although statistically significant association was not detected for individual SNPs in the NIMH data set, the same haplotype was consistently overtransmitted in both series. These data suggest that a susceptibility variant for bipolar illness exists in the vicinity of the G72/G30 genes. Taken together with the earlier report, this is the first demonstration of a novel gene(s), discovered through a positional approach, independently associated with both bipolar illness and schizophrenia. PMID- 12647259 TI - Genome association studies of complex diseases by case-control designs. AB - One way to perform linkage-disequilibrium (LD) mapping of genetic traits is to use single markers. Since dense marker maps-such as single-nucleotide polymorphism and high-resolution microsatellite maps-are available, it is natural and practical to generalize single-marker LD mapping to high-resolution haplotype or multiple-marker LD mapping. This article investigates high-resolution LD mapping methods, for complex diseases, based on haplotype maps or microsatellite marker maps. The objective is to explore test statistics that combine information from haplotype blocks or multiple markers. Based on two coding methods, genotype coding and haplotype coding, Hotelling's T2 statistics TG and TH are proposed to test the association between a disease locus and two haplotype blocks or two markers. The validity of the two T2 statistics is proved by theoretical calculations. A statistic TC, an extension of the traditional chi2 method of comparing haplotype frequencies, is introduced by simply adding the chi2 test statistics of the two haplotype blocks together. The merit of the three methods is explored by calculation and comparison of power and of type I errors. In the presence of LD between the two blocks, the type I error of TC is higher than that of TH and TG, since TC ignores the correlation between the two blocks. For each of the three statistics, the power of using two haplotype blocks is higher than that of using only one haplotype block. By power comparison, we notice that TC has higher power than that of TH, and TH has higher power than that of TG. In the absence of LD between the two blocks, the power of TC is similar to that of TH and higher than that of TG. Hence, we advocate use of TH in the data analysis. In the presence of LD between the two blocks, TH takes into account the correlation between the two haplotype blocks and has a lower type I error and higher power than TG. Besides, the feasibility of the methods is shown by sample-size calculation. PMID- 12647260 TI - Antihypertensive treatment and homocysteine concentrations. AB - Thiazides and angiotensin-converting enzyme (ACE) inhibitors are first-choice drugs for lowering elevated blood pressure and hence risk of cardiovascular disease. Homocysteine (tHcy) is another and independent cardiovascular risk factor and has been reported to be elevated in patients on antihypertensive therapy. As these studies reported only associations, a preliminary, randomized, prospective treatment study was performed in 40 hypertensive patients. We investigated the major determinants of tHcy concentrations after treatment with hydrochlorothiazide (HCT) or captopril: vitamins B6, B12, folic acid, and creatinine and cystatin C as parameters of renal function. A total of 21 Patients were treated with HCT and 19 with captopril, for, respectively, 31 and 29 days. HCT, but not captopril, raised tHcy by 16% (P =.003) and also creatinine and cystatin C (P =.025 and P =.004, respectively). This tHcy increase may offset the desired cardioprotection conferred by lowering the blood pressure. PMID- 12647261 TI - Chronic effects of dehydroepiandrosterone on rat adipose tissue metabolism. AB - The goal of the present study was to examine cellular mechanisms that regulate adipose cell metabolism in ovariectomized (OVX) and intact rats that were subjected to long-term (27 weeks) treatment with dehydroepiandrosterone (DHEA). Forty-eight 16-month-old female rats were divided into 4 groups of 9 to 11 animals (intact, intact-DHEA, OVX, OVX-DHEA). Adipose tissue lipoprotein lipase (LPL), hormone-sensitive lipase (HSL), and cyclic adenosine monophosphate (cAMP) dependent phosphodiesterase (cAMP-PDE) activities were determined, and alpha2-, beta1/beta2-, and beta3-adrenoceptors (ARs) were quantified. DHEA did not affect body weight, fat, or muscle mass in intact rats. The similar retroperitoneal fat pad weight of intact-DHEA rats compared to intact animals was in agreement with the lack of difference in the enzyme activities and AR densities. The increased body weight of OVX rat was paralleled by a greater retroperitoneal adipose tissue mass (P <.01), which was in turn associated with a marked rise in LPL activity (P <.005) and a slight decrease in HSL activity (P <.05) compared to intact animals. OVX-DHEA rats, compared to untreated OVX animals, had a smaller retroperitoneal fat depot, which correlated with a decrease in LPL activity (P <.005) and moderate increase in both HSL activity and beta3-AR density (P <.05). DHEA treatment lowered fasting insulin and triglyceride levels in both intact and OVX rats (P <.05). Plasma testosterone, androsterone, androstenedione, and androstenediol levels were also significantly increased in both intact-DHEA and OVX-DHEA rats compared to untreated animals (P <.0001). These findings suggest that the antiobesity action of DHEA may be related in part to changes in lipase activities and in beta3-AR density, and that it is dependent on the ovarian status of the animal. PMID- 12647262 TI - Evidence for a direct effect of captopril on early steps of insulin action in BC3H-1 myocytes. AB - Captopril, an angiotensin-converting enzyme (ACE) inhibitor, has been reported to improve insulin sensitivity. However, despite extensive investigation, the mechanisms responsible for this effect are not fully understood. Reduction of plasma angiotensin II and inhibition of kininase II have been suggested to contribute to improve insulin sensitivity. Insulin binding was measured at tracer insulin concentration in intact cells with or without captopril treatment. Specific binding, expressed as percent of total insulin added, was not different in control and captopril-treated cells. However, captopril treatment caused an increase in insulin-induced insulin receptor substrate-1 (IRS-1) phosphorylation accompanied by an increased association of IRS-1 with phosphoinositide-3 kinase (PI-3 kinase), despite no change on insulin receptor (IR) autophosphorylation. There was also an increased threonine kinase B (AKT) phosphorylation in captopril treated cells followed by enhanced basal and insulin-stimulated glucose uptake. These results indicate that captopril treatment has a direct effect on early phosphorylation events induced by insulin in BC3H-1 myocytes. PMID- 12647263 TI - Interactions of apolipoprotein E genotype and dietary fat intake of healthy older persons during mid-adult life. AB - In a case control study of genetic and lifestyle risk factors for Alzheimer's disease (AD), we obtained recalled food consumption frequencies translated to nutrients and averaged over 2 age periods of adult life, 20 to 39 and 40 to 59 years. The proportion of controls with the apolipoprotein E epsilon4 (APOE epsilon4) genotype was significantly higher in the lowest tertile of fat consumption (36.3% of energy) compared with controls with epsilon4 in the highest tertile of fat intake (44.6% of energy). Healthy older persons with the epsilon4 allele who survived to be included in this study may be protected with lower dietary fat intake and other healthy behaviors. Diet-genotype interactions may have important influences on disorders of later life. PMID- 12647264 TI - Cholesteryl ester transfer protein and lecithin:cholesterol acyltransferase activities in hispanic and anglo postmenopausal women: associations with total and regional body fat. AB - Reverse cholesterol transport is one process by which high-density lipoprotein (HDL) cholesterol has been hypothesized to play a role in reducing the risk of coronary heart disease. This study was designed to examine cholesteryl ester transfer protein (CETP) and lecithin:cholesterol acyltransferase (LCAT) activities, 2 modulators of reverse cholesterol transport, in Hispanic and Anglo postmenopausal women. The associations between plasma CETP and LCAT activities and body composition were also examined. Of the 199 subjects, 33% were of Hispanic origin and 47% were undergoing hormone replacement therapy (HRT). Body composition was measured by dual-energy x-ray absorptiometry (DXA) and anthropometry. Plasma CETP activity was higher in Hispanic compared to Anglo women, although the difference was eliminated when data were adjusted for abdominal fat. Hispanic women had lower plasma HDL cholesterol concentrations, higher total cholesterol:HDL cholesterol ratios and triglyceride concentrations, and greater susceptibility of low-density lipoprotein (LDL) particles to oxidation. Hispanic women also had a significantly greater relative deposition of body fat on the trunk and intra-abdominally than did Anglo women, even after adjusting for total body fat. There were no significant ethnic differences in LCAT activity. Plasma CETP and LCAT activities were negatively correlated with HDL cholesterol and positively correlated with total cholesterol, LDL cholesterol, and triglycerides, as well as total and regional body composition variables. In conclusion, results suggest a greater risk for coronary heart disease in Hispanic women compared to Anglo women. PMID- 12647265 TI - Angiotensin II upregulates the expression of vasopressin V2 mRNA in the inner medullary collecting duct of the rat. AB - Previous in vivo studies in cardiomyopathic hamsters suggested that the expression of vasopressin (AVP) V2 mRNA is up- regulated by angiotensin II. The present study was performed to determine whether angiotensin II plays a role in regulating the expression of AVP V2 mRNA and aquaporin-2 (AQP2) mRNA in the inner medullary collecting duct (IMCD) of the male Wistar rat. The expression of AVP V2 mRNA and AQP2 mRNA in the IMCD was measured by competitive reverse-transcriptase polymerase chain reaction (RT-PCR). Six groups of experiments were performed. In the first group, we incubated IMCD with 3 different doses of angiotensin II (10( 11), 10(-9) and 10(-7) mol/L). Angiotensin II caused a significant increase in the AVP V2 mRNA in a dose-dependent manner but its effect on AQP2 mRNA was modest. This effect of angiotensin II was inhibited by angiotensin II receptor antagonist, [Sar1,Ile8]-angiotensin II. To examine the role of PKA in mediating an increase in AVP V2 mRNA expression, we incubated IMCD with 10(-7) and 10(-11) M of angiotensin II in the presence of a specific protein kinase A (PKA) inhibitor, Rp diasteroisomer of adenosine 3'-5'-cylic monophosphothionate (Rp cAMPS). The angiotensin II-induced upregulation of V2 mRNA was abolished. In the fourth group, we examined the effect of protein kinase C (PKC) inhibition on V2 mRNA expression. The upregulation of V2 mRNA induced by angiotensin II was greatly exaggerated when IMCD was incubated with angiotensin II and RO-31-8220 (PKC inhibitor). In the fifth and sixth groups of studies, we determined the direct effect of PKA and PKC on regulating the expression of V2 mRNA and AQP2 mRNA in the IMCD, respectively. Dibutryl cAMP stimulated an upregulation in the expression of V2 mRNA and AQP2 mRNA, whereas phorbol esters suppressed the expression of V2 mRNA. These results suggested that PKA stimulates and PKC suppresses the expression of V2 mRNA in the IMCD of the kidney. PMID- 12647266 TI - Evidence for metabolic and endocrine abnormalities in subjects recovered from anorexia nervosa. AB - Subjects with anorexia nervosa (AN) at low weight display metabolic, endocrine, and behavioral abnormalities. Whether these various differences are a consequence of the condition and persist after recovery is unclear. We tested the hypothesis that abnormalities in the insulin and leptin axes and in the desire to eat persisted in subjects who had recovered from AN in terms of body mass index (BMI) and menstrual function. Endocrine, metabolic, and psychological parameters were assessed by sampling under fasting conditions and serially in response to a standard meal. Subjects included 18 females recovered from AN and 18 female controls and measures included plasma insulin, leptin, glucose and beta hydroxybutyrate (beta-HBA) concentrations together with desire to eat. Fasting glucose concentrations were normal in both groups, but fasting insulin concentrations were significantly lower and the fasting glucose/insulin ratio significantly higher in the recovered subjects. The glucose concentration was significantly higher at the end of the meal period in the recovered group. The peak increase of insulin during the meal was significantly less in the recovered group and in response to the meal, glucose/insulin ratios were significantly higher for the first 45 minutes indicating a delayed insulin response. Fasting beta-HBA concentrations were not significantly different between groups, but postmeal decreases were significant and larger in the recovered AN group. Fasting and meal-related leptin concentrations were not significantly different between the groups and in both groups were correlated with BMI. In controls, but not in recovered subjects, the reported desire to eat was correlated with plasma glucose and leptin concentrations. The insulin, glucose and beta-HBA data indicated the presence of insulin hypersensitivity in the recovered subjects. As the insulin response to the meal was blunted and apparently delayed, there may be a persistent alteration in pancreatic function as a long-term pathological consequence of the anorexia. Alternatively, these data indicate a possible trait marker for AN. PMID- 12647267 TI - Plasma concentrations of asymmetric-dimethyl-arginine in type 2 diabetes associate with glycemic control and glomerular filtration rate but not with risk factors of vasculopathy. AB - Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide synthase (NOS). Increased plasma levels of ADMA may indicate endothelial dysfunction and increased risk of angiopathy. The relation of ADMA to diabetes, glycemic control, and renal function, especially early diabetic hyperfiltration, remains unknown. We tried to evaluate whether there is an association between ADMA and glycosylated hemoglobin (GHbA(1c)) on the one hand and glomerular filtration rate (GFR) on the other hand in diabetic subjects with normal or slightly increased GFR. We also studied whether plasma ADMA is associated with some risk factors of vasculopathy (hypercholesterolemia and hypertension). The study subjects consisted of 86 patients with type 2 diabetes and 65 control subjects. Plasma ADMA levels were measured by high-pressure liquid chromatography as o-pthalaldehyde (OPA) derivatives and GFR was determined by the plasma clearance of chromium 51-EDTA. The diabetic patients had lower plasma ADMA levels than the nondiabetic control subjects (0.29 +/- 0.15 v 0.34 +/- 0.16 micromol/L, P <.03). In the diabetic subjects, plasma ADMA concentrations were inversely correlated with GHbA(1c) (R = -0.28, P =.01). In a multivariate linear model, significant predictors of ADMA were GFR (R = -0.32, P =.008) in diabetic subjects and GHbA(1c) (R = -0.19, P =.03) and GFR (R = -0.19, P =.02) in all subjects. Plasma ADMA was not associated with risk factors of vasculopathy. We conclude that diabetic patients with a normal or slightly increased GFR have lower circulating ADMA concentrations than nondiabetic control subjects. In type 2 diabetic patients high GFR and poor glycemic control were related to low plasma ADMA concentrations. PMID- 12647268 TI - Atherogenic role of lysophosphatidylcholine in low-density lipoprotein modified by phospholipase A2 and in diabetic patients: protection by nitric oxide donor. AB - The aim of our study was to investigate the atherogenic role of lysophosphatidylcholine (lyso-PC) in low-density lipoprotein (LDL) under diabetic environment. Expression of monocyte chemoattractant protein-1 (MCP-1) mRNA and nuclear factor-kappa B (NF-kappaB)-DNA binding activity were determined in human umbilical vein endothelial cells (HUVEC) incubated with native or glycoxidized LDL, LDL modified by phospholipase A2 (PLA2) and LDL isolated from diabetic patients. Lyso-PC contents in LDL were measured using electrospray ionization liquid chromatography/mass spectrometry (ESI-LC/MS). Lyso-PC contents were higher in glycoxidized LDL and PLA2-treated LDL compared with native LDL. Glycoxidized LDL and enrichment of lyso-PC by PLA2 treatment resulted in upregulation of MCP-1 mRNA expression through increased NF-kappaB activity in HUVEC. Moreover, LDL isolated from diabetics contained more lyso-PC than that from nondiabetic subjects, and induced higher MCP-1 mRNA expression and NF-kappaB activity in HUVEC. In both in vitro and human studies, palmitoyl- and stearoyl-lyso-PC contents correlated with MCP-1 expression and NF-kappaB activity. Preincubation with 4-ethyl-2-hydroxyimino-5-nitro-3-hexenamide, a NO donor, abrogated increased expression of MCP-1 mRNA and high NF-kappaB activity induced by PLA2-treated LDL and by LDL isolated from diabetic patients. Our results suggest that lyso-PC contents in LDL play an important role in atherogenesis under diabetic condition, which could be prevented by increased availability of vascular NO. PMID- 12647269 TI - Plasma galanin response to head-up tilt in normal subjects and patients with recurrent vasovagal syncope. AB - Neurohumoral factors may contribute to cardiovascular changes associated with vasovagal syncope (VVS). Galanin (GAL) is a neuropeptide, widely distributed in the central and peripheral nervous systems, that interacts with both sympathetic and vagal systems as well as with neurotransmitters, such as serotonin. We investigated the changes in plasma GAL and catecholamine levels during head-up tilt (HUT) test in patients with recurrent VVS. Twenty-two patients (11 women, aged 33.1 +/- 4.2 years) with a history of VVS and 10 healthy subjects (5 women, aged 38.0 +/- 5.8 years) underwent HUT test (60 degrees, 45 minutes). GAL and catecholamine plasma levels were measured in the supine position, during HUT and, in patients with positive response, at presyncope, syncope, and after recovery of consciousness. Thirteen patients developed syncope during HUT, whereas no healthy subjects had a positive response. In healthy subjects, GAL did not change during HUT. By contrast, in patients with a history of VVS and a negative response to tilting (no syncope), GAL significantly (P <.001) increased in response to tilting (supine, 10.2 +/- 0.6 pmol/L; tilting, 18.1 +/- 1.1 pmol/L at 45 minutes) and correlated positively with the increases in blood pressure (BP) and heart rate (HR). In patients with a positive response, GAL did not change either before the loss of consciousness or during syncope. In patients with a positive response, norepinephrine (NE) significantly (P <.001) increased during tilting and then remained practically unchanged during syncope, whereas epinephrine (E) significantly (P <.001) increased during tilting and then showed further significant increases at presyncope and syncope. In conclusion, this study shows that circulating GAL levels progressively increase in correlation with the cardiovascular parameters during a negative HUT in patients with a history of VVS, whereas they remain unchanged in healthy subjects. Moreover, in the patients with tilting-induced syncope GAL does not change either before or during loss of consciousness. These data suggest a role for endogenous GAL in the adaptive responses to acute orthostatic stress preventing syncope in susceptible individuals. PMID- 12647270 TI - Effect of food restriction on lactate sarcolemmal transport. AB - The objective of this study was to investigate the effects of 6 weeks of food restriction (FR) on sarcolemmal lactate transport in rats. The daily food consumption of rats was monitored for 10 days, after which they were assigned to either a control group (CTL, n = 7) that consumed food ad libitum or an FR group (n = 7) that received a daily ration equal to 60% of their predetermined baseline food intake. After the 6-week period, we observed in red gastrocnemius (RG) a fall of 48% in glycogen content (P <.01) and a reduction in glutathione peroxidase activity (P <.05), confirming that the FR program was well executed. FR resulted in a reduction in muscle lactate (P <.05) and liver glycogen contents (P <.01). Moreover, hyperlactatemia was noted in the FR group: 1.77 +/- 0.24 versus 2.67 +/- 0.29 mmol/L (P <.05). Lactate transport capacity was significantly increased (P <.05) in FR rats, although monocarboxylate transporter isoforms (MCT1 and MCT4) did not change significantly. We conclude that FR alters sarcolemmal lactate transport activity without affecting MCT1 and MCT4 expression. PMID- 12647271 TI - Early vascular damage in primary hypoalphalipoproteinemia. AB - The relationship between hypoalphalipoproteinemia (hypoalpha), a metabolic disorder characterized by reduced high-density lipoprotein (HDL) cholesterol levels, and atherosclerotic disease is not completely understood. We investigated arterial functional and structural changes in 19 subjects with hypoalpha (HDL cholesterol < or = 0.7 mmol/L for men and < or = 0.8 mmol/L for women; 13 men; 47 +/- 7 years) and in 21 healthy control subjects (11 men; 46 +/- 13 years). Brachial-artery flow-mediated vasodilation (FMV) and intima-media thickness (IMT) of the carotid and femoral arteries were determined in all subjects. FMV was significantly lower in hypoalpha than in controls (5.6% +/- 4.3% v 8.2% +/- 2.7%; P <.05). IMT was greater in hypoalpha than in controls at both the internal carotid (0.83 +/- 0.1 mm v 0.69 +/- 0.1 mm) and superficial femoral level (0.83 +/- 0.2 mm v 0.68 +/- 0.1 mm; both P <.05). FMV had a positive correlation with HDL cholesterol (r =.42, P =.06) and a negative one with triglycerides (r = 0.38, P =.01). An inverse relationship was found between HDL cholesterol and internal carotid and superficial femoral IMT (r = -0.64 and r = -0.60, respectively; P <.01 for both) and a positive one between triglycerides and internal carotid and superficial femoral IMT (r =.53 and r =.47, P <.05). In a multivariate regression analysis, brachial FMV was predicted by HDL cholesterol and brachial diameter (beta =.42 and -0.43, respectively; both P <.05). HDL cholesterol was the only significant predictor of internal carotid and superficial femoral IMT (beta = -0.45 and -0.49, respectively; both P <.05). In conclusion, subjects with primary hypoalpha, without overt cardiovascular disease, are characterized by an impaired endothelial function and by an increase in large-artery IMT. PMID- 12647272 TI - Short-term dietary adjustment with a hydrolyzed casein-based diet postpones diabetes development in the diabetes-prone BB rat. AB - From earlier studies it appears that weaning associated changes in the animal's physiology and that of the pancreas in particular, render diabetes-prone Bio Breeding (DP-BB) rats susceptible to the induction and development of insulin dependent diabetes mellitus (IDDM). In this study we tested whether a short-term dietary adjustment at weaning would influence the development of diabetes later in life. For this purpose a diet in which the protein source was replaced with hydrolyzed casein (HC) was given to the rats from weaning to 60 days of age and from weaning to 130 days of age. The control group received the cereal-based standard diet throughout the experiment. The short-term dietary adjustment resulted in a significant delay of diabetes development. The rats fed the HC diet from weaning to 130 days of age showed a lower incidence of diabetes at 130 days of age. No differences were seen in the histological insulitis scores between the rats of the different treatment groups. Interestingly, when testing (mucosal) immune functions of short-term HC-fed rats, their mesenteric lymph node cells (MLNC) showed increased interferon-gamma (IFN-gamma) and reduced interleukin-10 (IL-10) production after in vitro stimulation. These results demonstrate that short-term dietary adjustments at a young age can influence the course of diabetes later in life. The shift in cytokine profile of MLNC of the HC-fed rats suggests that mechanisms involved can be at the level of both the (mucosal) immune system and the beta cell. PMID- 12647273 TI - The lipoprotein lipase gene HindIII polymorphism is associated with lipid levels in early-onset type 2 diabetic patients. AB - Lipoprotein lipase (LPL) plays a central role in triglyceride metabolism, and the LPL gene T495G HindIII polymorphism has been associated with variations in lipid levels and heart disease in Caucasians with the more common H+ allele being associated with adverse lipid profiles and increased risk of CHD. We investigated this polymorphism in 785 Chinese subjects with varying components of the metabolic syndrome, including 61.4% with early-onset type 2 diabetes (age at diagnosis < or = 40 years), and 167 healthy control subjects using a polymerase chain reaction (PCR)-based restriction fragment length polymorphism (RFLP) method. The allele and genotype frequencies were similar in the patients and control subjects. When grouped above or below standard cutoffs for triglyceride levels, the H+ allele was more frequent in hypertriglyceridemic than that in normotriglyceridemic subjects in the total population (81.5% v 76.1%) and early onset type 2 diabetics (84.4% v 77.4%, both P <.05). Moreover, H+H+ carriers had significantly higher plasma triglyceride and lower high-density lipoprotein (HDL) cholesterol levels when compared to subjects with the H- allele in the total population, and in patients with early-onset diabetics (both P <.05). In the total population and the early-onset diabetic patients, this relationship was confined to males when gender was considered. We conclude that the H+ allele of the LPL gene HindIII polymorphism is associated with higher plasma triglyceride and lower HDL-cholesterol levels in Chinese patients with early-onset diabetes. PMID- 12647274 TI - In vivo induction of insulin secretion by ornithine alpha-ketoglutarate: involvement of nitric oxide and glutamine. AB - We previously demonstrated that ornithine alpha-ketoglutarate (OKG), known for its anabolic properties, induces insulin secretion in vitro. The present study was undertaken to further characterize this effect in vivo and investigate a possible interaction with glucose both in vivo and in vitro. Male Wistar rats received an intravenous bolus of OKG (25 mg/kg) and/or glucose (0.8 g/kg) or saline, and their plasma insulin and glucose levels were monitored for 30 minutes. OKG alone increased plasma insulin to a similar extent to glucose. In combination with glucose, OKG significantly increased glucose-induced insulin secretion in vivo and in vitro, and led to a significant increase in glucose utilization in vivo. The absence of significant variations in plasma arginine and glutamine suggests a direct effect of OKG on the pancreas. To assess the involvement of the synthesis of nitric oxide and glutamine in OKG-induced insulin secretion, the experiments were repeated in the presence of inhibitors of these 2 pathways, respectively L-nitroarginine-methylester (L-NAME) and methionine sulfoximine (MSO). Both inhibitors were able significantly to reduce OKG-induced insulin secretion without affecting either basal or glucose-induced insulin release. Thus OKG acts directly with glucose on islets to induce insulin secretion via mechanisms involving NO and glutamine synthesis. In addition, our results suggest that OKG and glucose act via separate pathways. PMID- 12647275 TI - Reductions in plasma cholesterol levels after fenofibrate treatment are negatively correlated with resistin expression in human adipose tissue. AB - The adipocyte-derived cytokine, resistin, has been proposed as the link between obesity and type 2 diabetes mellitus in murine models. In humans, resistin is identical to FIZZ3 (found in inflammatory zone 3), which belongs to a family of proteins that appears to be involved in inflammatory processes. To study the mechanisms by which fibrates improve glucose homeostasis, we determined resistin mRNA levels by using relative quantitative reverse-transcriptase-polymerase chain reaction (RT-PCR) in omental white adipose tissue samples obtained from patients treated with placebo or fenofibrate (200 mg/d) for 8 weeks before elective cholecystectomy. Fenofibrate treatment reduced total plasma cholesterol and low density lipoprotein (LDL)-cholesterol levels by 24% and 35%, respectively. Compared with placebo values, a 2.4-fold induction in resistin mRNA levels was observed in white adipose tissue of fenofibrate-treated patients, whereas no changes were observed in the mRNA levels of the well-known perosixome proliferator-activated receptor (PPAR) target genes CD36, acyl-CoA oxidase, and carnitine palmitoyltransferase. These findings indicate that resistin changes were not related to PPAR activation by fenofibrate. Interestingly, resistin mRNA levels showed a negative correlation with plasma cholesterol levels (r2 =.53, P =.039, n = 8), but not with triglyceride levels (r2 =.02, P =.73, n = 8). These results suggest that cholesterol regulates resistin expression in human white adipose tissue. PMID- 12647276 TI - The polymorphism of the beta3-adrenergic receptor gene is associated with reduced low-density lipoprotein particle size. AB - People with a predominance of small, dense low-density lipoprotein (LDL) particles appear to be at increased risk for coronary disease, independent of LDL cholesterol levels. The Trp64Arg variant of the beta3-adrenergic receptor gene is reported to be associated with abdominal obesity and resistance to insulin, and as a consequence, this variant may be a genetic factor in the development of atherosclerosis. Therefore, we investigated whether the beta3-adrenergic receptor polymorphism contributes to the distribution of LDL particle size in 136 Japanese subjects, aged 33 to 59 years, who visited for a routine annual checkup. None of these subjects were taking any medication. The diameter of LDL particles was determined at their peak size using nondenaturing 2% to 16% polyacrylamide gradient gels using fresh plasma samples. The genotype frequencies were: Trp/Trp, 71.3%; Try/Arg, 22.1%; and Arg/Arg, 6.6%, with allele frequencies of 0.82 for Trp64 and 0.18 for Arg64. The subjects with the Arg/Arg genotype had significantly higher levels of fasting plasma insulin and triglycerides and an insulin resistance index of homeostasis model assessment (HOMA-R), and significantly smaller LDL particle size than did the subjects with the Trp/Trp genotype. After adjusting for fasting insulin, body mass index (BMI), and HOMA-R index, there was no longer an observed difference in LDL particle size. The number of the Arg64 allele in individuals was significantly related with fasting insulin, BMI, triglycerides, glycosylated hemoglobin (HbA1c), and fasting glucose, and it was inversely related with LDL particle size. After adjusting for triglyceride, fasting insulin levels, and HOMA-R index, LDL particle size was no longer inversely correlated with the Arg allele. These findings suggest that the Trp64Arg variant in the beta3-adrenergic receptor gene may be associated with reducing LDL particle size, probably due to insulin resistance. PMID- 12647277 TI - Risk factor clustering in the insulin resistance syndrome and its relationship to cardiovascular disease in postmenopausal white, black, hispanic, and Asian/Pacific Islander women. AB - The aim of this study was to examine how major components of the insulin resistance (IR) syndrome relate to each other and to cardiovascular disease (CVD) in postmenopausal women in 4 ethnic groups. Baseline data from the Women's Health Initiative (WHI) on 3,083 50- to 79-year-old women (1,635 white, 802 black, 390 Hispanic, and 256 Asian/Pacific Islander) were examined. Participants underwent a personal interview and a physical examination, blood samples were drawn, and a detailed cardiovascular history was ascertained. Factor analysis was used to assess the clustering and interdependence of groups of CVD-related IR syndrome variables. Four factors were identified. An obesity factor included IR in all groups and had a significant association with CVD in white (P =.0001) and Hispanic (P =.0024) women. A dyslipidemia factor (high-density lipoprotein [HDL], triglycerides, and HDL2: total HDL ratio) also included insulin and IR and was significantly correlated with CVD in black (P=.0006) and Hispanic (P =.0217) women and had a borderline association in white women (P =.068). Total and low density lipoprotein (LDL) cholesterol did not relate to CVD in any group. Blood pressure was related weakly to CVD in white women (P =.0434) and strongly in black women (P =.0095). Components of the IR syndrome appear to be associated with CVD in postmenopausal women, although the magnitude of these relationships differed by ethnicity. PMID- 12647278 TI - Antihyperglycemic and blood pressure-reducing effects of stevioside in the diabetic Goto-Kakizaki rat. AB - Stevioside, a glycoside present in the leaves of the plant, Stevia rebaudiana Bertoni (SrB), has acute insulinotropic effects in vitro. Its potential antihyperglycemic and blood pressure-lowering effects were examined in a long term study in the type 2 diabetic Goto-Kakizaki (GK) rat. Rats were fed 0.025 g x kg(-1) x d(-1) of stevioside (purity > 99.6%) for 6 weeks. An intra-arterial catheter was inserted into the rats after 5 weeks, and conscious rats were subjected to arterial glucose tolerance test (2.0 g x kg(-1)) during week 6. Stevioside had an antihyperglycemic effect (incremental area under the glucose response curve [IAUC]): 985 +/- 20 (stevioside) versus 1,575 +/- 21 (control) mmol/L x 180 minutes, (P <.05), it enhanced the first-phase insulin response (IAUC: 343 +/- 33 [stevioside] v 136 +/- 24 [control] microU/mL insulin x 30 minutes, P <.05) and concomitantly suppressed the glucagon levels (total AUC: 2,026 +/- 234 [stevioside] v 3,535 +/- 282 [control] pg/mL x 180 minutes, P <.05). In addition, stevioside caused a pronounced suppression of both the systolic (135 +/- 2 v 153 +/- 5 mm Hg; P <.001) and the diastolic blood pressure (74 +/- 1 v 83 +/- 1 mm Hg; P <.001). Bolus injections of stevioside (0.025 g x kg(-1)) did not induce hypoglycemia. Stevioside augmented the insulin content in the beta-cell line, INS-1. Stevioside may increase the insulin secretion, in part, by induction of genes involved in glycolysis. It may also improve the nutrient-sensing mechanisms, increase cytosolic long-chain fatty acyl-coenzyme A (CoA), and downregulate phosphodiesterase 1 (PDE1) estimated by the microarray gene chip technology. In conclusion, stevioside enjoys a dual positive effect by acting as an antihyperglycemic and a blood pressure-lowering substance; effects that may have therapeutic potential in the treatment of type 2 diabetes and the metabolic syndrome. PMID- 12647280 TI - [Considerations on continuing medical education]. AB - How to prepare questionnaires for continuing medical education. How to prepare meetings and talks. How to present slides at meetings. These three topics are very important for the success of the continuing medical education. Chairmen and speakers involved in the education of young physicians or in the communication of new information on life sciences should be well acquainted with the points listed above. PMID- 12647279 TI - Serum concentrations of dehydroepiandrosterone sulfate and leptin in obese patients with normal serum cholesterol. AB - Normal (< 200 mg/dL) serum concentrations of cholesterol and a favorable ratio of cholesterol/high-density lipoprotein (HDL)-cholesterol are frequently seen in morbidly obese (body mass index [BMI] > 35 kg/m2) patients. Because it is unknown whether this subgroup is characterized by differences in other potential markers of cardiovascular disease, serum concentrations of dehydroepiandrosterone sulfate (DHEAS) and leptin were determined in 155 obese patients (BMI > 35 kg/m2, aged 20 to 50 years) with normal (n = 72) or with elevated (n = 83) total serum cholesterol. We found that seemingly negative marginal correlations between serum concentrations of DHEAS and cholesterol, as well as between DHEAS and the ratio cholesterol/HDL-cholesterol, were not any more apparent after correction for age, sex, and BMI. A negative correlation between serum leptin concentrations and the ratio cholesterol/HDL-cholesterol persisted after correction for age, sex, and BMI. In morbid obesity, there appears to be an association between serum concentrations of leptin and a more favorable lipid profile, whereas there is no direct interrelation between serum concentrations of cholesterol and DHEAS. PMID- 12647281 TI - [Renal damage in type 2 diabetes]. AB - Renal involvement in patients with type 2 diabetes will (probably) be one of the most important clinical problems for nephrologists to face during the next few years. Unlike type 1 diabetes, in type 2 diabetes the renal damage has not yet been well defined at both clinical and pathological levels. Pathological examination of renal biopsies has displayed different patterns of renal damage including diabetic glomerulosclerosis (Class 1), mostly chronic vascular changes (Class 2) and superimposed glomerular diseases (Class 3a) or unrelated to diabetic glomerulosclerosis (Class 3b). Despite the large number of papers published in this field, the actual prevalence and outcome of the different histological classes still remain to be established. Reported discrepancies are most likely caused by ethnic and geographic factors. However, as documented by a recent study carried out on a large number of patients, the prevalence of histological patterns is also greatly influenced by the policy for performing renal biopsies adopted at the various nephrological centers. Although the natural history of type 2 glomerulosclerosis (Class 1) still remains to be defined, those patients with clinical nephropathy and impairment of renal function have very poor outcome with a high rate of mortality and progression to uremia. Moreover, when diabetic glomerulosclerosis is complicated by superimposed glomerular diseases (Class 3a) the prognosis is much worse. On the contrary, when glomerular diseases are not associated with glomerulosclerosis lesions (Class 3b) the prognosis is markedly better. During the last ten years controlled studies have shown that the outcome in type 1 diabetic nephropathy has improved as a result of the use of drugs inhibiting the renin-angiotensin system. Although it is likely that this type of drug might also favourably influence the outcome of type 2 diabetic nephropathy, any conclusive evidence is presently still lacking. PMID- 12647282 TI - [Physiopathology of pre-eclampsia]. AB - BACKGROUND: Pre-eclampsia is a pregnancy-specific disorder that presents major health problems for both mothers and babies. It is a major cause of maternal mortality, especially in developing countries, where it may account for 80% of maternal deaths. In this review we have discussed the putative pathogenetic mechanisms of this severe illness. METHODS: Review of the international literature on pre-eclampsia. RESULTS: Pre-eclampsia occurs predominantly in primigravidas and is associated with increased foetal and maternal risk. Generally speaking, one could hypothesise that during pre-eclampsia the oxidative, coagulative and vasomotor balance is altered by increased sensitivity to angiotensin II associated with reduced synthesis of vasodilator prostaglandins, sympathetic nervous system hyperactivity, hyperlipidaemia associated with raised lipoperoxide synthesis and incomplete implant of cytotrophoblast in maternal spiral arteries. General vasoconstriction could then be a consequence with related utero-placental ischaemia and endothelial damage. These features are clinically characterised by arterial hypertension, proteinuria and sodium retention. The predisposition to develop pre-eclampsia seems to be at least partially hereditary and the risk seems to be transmissible through male sons. To explain the susceptibility of some women to develop pre-eclampsia, specific genetic polymorphisms have been associated to increased risk. CONCLUSIONS: Many factors are involved in the pathogenesis of pre-eclampsia. Occasionally these factors seem to act together, sometimes they do not and, consequently, it might be difficult to postulate a common pathogenesis for the disorder. Good obstetrical practice will identify known risk factors but will not enable physicians to determine the risk value for a particular pregnancy. Moreover, we are far from understanding the exact mechanisms involved, let alone acting upon them. The achievement of these objectives constitute a remarkable challenge for research. PMID- 12647283 TI - [Ethical problems in dialysis in elderly patients]. AB - The incidence and prevalence of elderly patients are progressively increasing in most dialysis facilities with consequent medical assistance difficulties and the need to find a suitable care unit. Particularly in this age group, the clinical assessment is often difficult and the selection criteria vary widely, not only from country to country, but also from one dialysis unit to another unit in the same area. The authors discuss some of the more complex arguments for and against dialysis in elderly patients. For some of the more difficult cases, Kantian deontology and its three ethical principles of beneficence, non-maleficence and autonomy can facilitate the decision-making process regarding the acceptance or refusal of the therapy. The central role of the patients themselves, the involvement of the family, the discussion of the individual case within the dialysis team, and the good performance of the pre-dialysis program are particularly important. In certain cases it is possible to actually discontinue the treatment, or not initiate it, by using different arguments in competent or incompetent patients. In conclusion, the difficult choice of whether to treat or turn down an elderly patient must depend exclusively on the medical clinical assessment of each case, whereas economical considerations can incorrectly influence that choice. PMID- 12647284 TI - [Role of iron deficiency in erythropoietin sensitivity in dialysis patients with elevated C-reactive protein]. AB - BACKGROUND: Chronic inflammation is a well-known cause of hyporesponsiveness of the bone marrow to erythropoietin (Epo). Factors which contribute to Epo resistance in the presence of inflammation include inhibition of erythroid precursor proliferation and functional iron deficiency induced by inflammatory cytokines. The specific role of iron deficiency in this clinical context, however, has not yet been clarified. METHODS: Our dialysis population consisted of 200 patients, from which 163 (91 males, mean age 67 +/- 12 years) who had been in dialysis for at least 4 (mean 62.4 +/- 71) months were selected for further study. Two groups were defined on the basis of C-reactive protein (CRP) concentrations: Group A (normal CRP < 5 mg/L; 78 patients) and Group B (elevated CRP > 15 mg/L; 43 patients). The remaining 42 patients with CRP in the range of 5 to 15 mg/L were excluded from the study. RESULTS: Erythropoietin dose and the parameter EpoDose/hemoglobin (Hb) were both greater in Group B (dose: 150 +/- 65 vs. 106 +/- 56 U/kg, p<0.001; EpoDose/Hb: 14.0 +/- 6 vs. 9.8 +/- 6, p<0.001. The two groups were stratified on the basis of transferrin saturation (tSAT) greater or less than 20%: A1 (tSAT > 20%, n = 52), A2 (tSAT < 20%, n = 26), B1 (tSAT > 20%, n = 19) and B2 (tSAT < 20%, n = 24). Erythropoietin dose and EpoDose/Hb were lower in A1 compared to A2 (dose: 96 +/- 52 vs. 124 +/- 6 U/kg, p<0.05; EpoDose/Hb: 8.4 +/- 5 vs. 12.4 +/- 7, p < 0.05), whereas in the B subgroups the variables were equally elevated (dose: 151 +/- 71 vs 142 +/- 59 U/kg, ns; EpoDose/Hb: 14.4 +/- 7 vs. 13.6 +/- 6, ns). Patients in subgroups A2 and B2 were treated with intravenous Fe gluconate 31 mg after each dialysis session for 6 months. Erythropoietin dose and EpoDose/Hb were significantly reduced only in subgroup A2 with normal CRP (dose: from 126 +/- 55 to 95 +/- 52 U/kg, p < 0.05; EpoDose/Hb: from 12.4 +/- 7 to 8.4 +/- 5, p < 0.05), whereas no improvement was observed in subgroup B2 with elevated CRP (dose: from 142 +/- 59 to 151 +/- 65 U/kg, ns; EpoDose/Hb: from 13.6 +/- 6 to 14.4 +/- 7, ns). CONCLUSIONS: Our data demonstrate that dialysis patients with CRP greater than 15 mg/L require an erythropoietin dose approximately 40% higher than patients with normal CRP, both in the presence and absence of iron deficiency. Iron therapy in patients with normal CRP and tSAT < 20% significantly improved the response to erythropoietin, but was completely ineffective in patients with increased CRP. These results suggest that functional iron deficiency plays a marginal role in resistance to erythropoietin observed in patients with elevated CRP concentrations. PMID- 12647285 TI - [Polyomavirus BK nephropathy in renal transplant: 2 cases with different clinical expressions and review of the literature]. AB - INTRODUCTION: Polyomavirus BK nephropathy is emerging as a significant cause of interstitial nephritis and allograft dysfunction (1-2). CASE REPORT: Two patients with renal transplants from cadaveric kidneys were treated with Tacrolimus plus Mycophenolate Mofetil (MMF) and Cyclosporine plus MMF, respectively. Their renal function gradually deteriorated eight to twelve months after the transplant. The renal biopsy of the first patient showed signs of significant interstitial tubulite, which necessitated the anti-rejection therapy with intravenous steroid pulses. After the pulses there was an additional dramatic increase in plasmatic creatinine, which suggested a revaluation of the kidney biopsy because of suspected Polyomavirus BK (BKV) nephropathy. In fact, after a more careful review, the suspicion of BKV infection was confirmed by the presence of intranuclear inclusions of tubular epithelium cells and marked denudation of the tubular basal membrane. The subsequent screening in both cases confirmed the presence of decoy cells in the urine, while the immunohistochemical analysis of the renal biopsy was strongly positive for the SV40 antigen. Our diagnosis was that of interstitial nephritis due to Polyomavirus BK that, in the first patient, was expressed by more aggressive clinical progress, probably due to enhanced immunosuppression from incorrect diagnosis of the interstitial rejection. The pre transplant clinical outcome of the first patient was characterised by proteinuric nephropathy without any histological confirmation. Furthermore, we observed abundant pre-transplant residual diuresis and glucose intolerance. All these elements led us to hypothesise that native kidneys could have a fundamental role as viral reservoirs. CONCLUSION: Even though we reconfirm the decisive role of the immunosuppressive therapy and of the donor s kidney as the fundamental causes of Polyomavirus reactivation, we believe that it cannot be the result of a possible active role by the native kidney. In fact, as already noted, the SV40 genome is important in the pathogenesis of focal gomerulosclerosis. Furthermore, reports of polyoma nephropathy in not-yet-transplanted patients could accredit the role of the native kidneys as important viral reservoirs capable of inducing nephropathy in renal transplant patients. PMID- 12647287 TI - [Complications of the nephrotic syndrome]. AB - We described the case of a 27-year-old man presenting pulmonary embolism and hyperlipidaemia. Subsequent investigation revealed that he was affected by renal vein thrombosis and nephrotic syndrome due to membranous glomeruloephritis. Nephrotic syndrome complications are numerous and may represent the first sign of the syndrome. Among these complications we find thromboembolism, infections, negative nitrogen balance and renal failure. There are very few prognostic indicators that enable the prediction of nephrotic syndrome complications. Recent advances in the understanding of alterations in the metabolism of circulating and somatic proteins associated with proteinuria and hypooncotic condition have led to new insights into the pathophysiologic processes associated with the syndrome. PMID- 12647286 TI - [Historical Archives of Italian Nephrology: the artificial kidney commissioned in 1947 by Aminta Fieschi (1904-1991)]. AB - Aminta Fieschi, a physician who is today remembered mainly for his contributions in the field of haematology, in the late 1940s devoted himself to the study of the treatment of uraemia. For this reason he commissioned, a manufacturer based in Milano, an artificial kidney, which was built on the design of the "rotating drum kidney" described by Willem Kolff in 1947 in his epoch-making monograph "New Ways of Treating Uraemia". However, after using that artificial kidney in few patients Fieschi abandoned it due to dangerous cardiovascular complications, and shifted to the use of other safer techniques such as intestinal perfusion. This paper describes the main contributions of Fieschi in the field of nephrology as well as the features of artificial kidney he used, whose manufacturer has been identified and retraced after more than fifty years by the author. PMID- 12647288 TI - [Spinal cord infarction during haemodialysis]. AB - A 26-year-old patient with chronic renal failure presented a spinal cord infarction during haemodialysis. This is the first case of a patient with chronic renal failure maintained on chronic haemodialysis described in literature. In this case, the severity of vascular lesions documented by widespread vascular calcifications were particularly striking. PMID- 12647289 TI - [Membranous glomerulonephritis and transitional cell carcinoma, improved proteinuria after each tumor resection]. AB - Nephrotic syndrome has been described in association with solid tumours, such as carcinoma of the lung or colon, whilst the relationship between urinary tract cancers and proteinuria is reported less frequently. We describe a case of a 75 year-old man with nephrotic syndrome and relapsing transitional cell carcinoma affecting the bladder. Renal biopsy showed membranous nephropathy. The patient underwent four transurethral tumour resections and each time we observed improved proteinuria by 3-4 weeks after surgery. On the contrary, steroid treatment was clearly unrelated to this improvement. We conclude that, at least in this case, surgical treatment of the tumour led to improved proteinuria whilst steroid treatment was not beneficial. PMID- 12647291 TI - Breast cancer gene 1 (BRCA1): role in cell cycle regulation and DNA repair- perhaps through transcription. AB - Mutations of BRCA1 gene are associated with more than half the cases of hereditary breast cancer. Breast cancer formation in BRCA1 mutation carriers is generally accompanied by loss of the wild-type allele, suggesting that BRCA1 protein may function as a tumor suppressor. The human BRCA1 gene encodes a nuclear protein of 1863 amino acids. Although several lines of evidences suggest that BRCA1 protein may have a role to play in cell cycle regulation, DNA repair, and other processes, the exact mechanism of functioning by BRCA1 protein is not clear. Recent evidences from several laboratories suggest that BRCA1 may regulate the expression of many genes like p21(WAF1/CIP1), Gadd45, Cyclin B1, DBB2, XPC, 14-3-3o and others at the level of transcription. These BRCA1-regulated gene products have been implicated directly or indirectly in cell cycle regulation and DNA repair. Thus a plausible model is proposed in which BRCA1 protein may bring its effects on cell cycle and DNA repair through its ability to modulate gene expression at the level of transcription. PMID- 12647290 TI - Bone morphogenetic protein-2 enhances osterix gene expression in chondrocytes. AB - Osterix is a recently identified zinc-finger-containing transcription factor, which is required for skeletogenesis as no bone formation was observed in osterix deficient mice. Osterix was first cloned as a gene whose expression was enhanced by BMP in C2C12 cells. As BMP induces ectopic bone formation in vivo via a pathway reminiscent to endochondral bone formation, BMP may also regulate osterix gene expression in chondrocytes. However, no information was available regarding the BMP actions on osterix gene expression in chondrocytes. We therefore examined the effects of BMP-2 on osterix gene expression in chondrocytes in culture. RT PCR analysis indicated that osterix mRNA was expressed in the primary cultures of chondrocytes derived from mouse rib cartilage. The treatment with BMP-2 enhanced the levels of osterix transcripts within 24 h and the enhancement was still observed at 48 h based on RT-PCR analysis. This BMP effect was specific to this cytokine, as TGF-beta did not alter osterix gene expression. BMP effects on the osterix mRNA levels were also confirmed by Northern blot analysis. The enhancing effect of BMP on osterix gene expression was observed in a dose-dependent manner starting at 200 ng/ml. The BMP enhancement of the osterix gene expression in chondrocytes was blocked in the presence of a protein synthesis inhibitor, cycloheximide, while it was still observed in the presence of 5,6-dichloro-1-beta D-ribofuranosylbenzimidazol (DRB) suggesting the involvement of post transcriptional events, which require new protein synthesis. These results indicated that osterix gene is expressed in the primary cultures of chondrocytes and its expression is under the control of BMP-2. PMID- 12647292 TI - Novel protein RGPR-p117: the gene expression in physiologic state and the binding activity to regucalcin gene promoter region in rat liver. AB - The binding activity of a novel regucalcin gene promoter region-related protein (RGPR-p117) to the TTGGC sequence of the rat regucalcin gene promoter region was investigated. The expression of RGPR-p117 mRNA was seen in the liver tissues of male and female rats. The sexual difference of this expression was not found. Liver RGPR-p117 mRNA expression was not changed with increasing age (1-50 weeks old), and its expression was not altered by fasting or refeeding. Nuclear factor I-A1 (NF1-A1) has been identified to be a transcription factor in stimulating the rat regucalcin gene promoter activity (Misawa and Yamaguchi [2002a] J Cell Biochem 84:795-802]. Recombinant nuclear factor I-A1 (NF1-A1) and RGPR-p117 proteins were used gel mobility shift assay. RGPR-p117 could not bind to TTGGC motif of the sequence between -525 and -504, which has been defined as a functional promoter element II-b. NF1-A1 was specifically bound to the II-b oligonucleotide. Moreover, RGPR-p117 was not bound to the II-b oligonucleotide in the presence of NF1-A1 or rat liver nuclear protein. The binding of NF1-A1 to the II-b oligonucleotide was not altered in the presence of RGPR-p117. This study demonstrates that RGPR-p117 mRNA, is expressed stably for physiologic change in rat liver, and that recombinant the protein does not directly bind to the TTGGC motif in rat regucalcin gene promoter. PMID- 12647294 TI - Type I collagen regulated dentin matrix protein-1 (Dmp-1) and osteocalcin (OCN) gene expression of rat dental pulp cells. AB - In this study, we investigated the effect of type I collagen on dentin matrix protein-1 (Dmp-1) and osteocalcin (OCN) gene expression of dental pulp cells. The mRNA level of Dmp-1 gene was down-regulated; however, OCN gene expression was up regulated by the culture of dental pulp cells with type I collagen. These findings imply that type I collagen regulates mRNA level of Dmp-1 and OCN gene that are predominantly expressed in active odontoblasts. The change of gene expression by type I collagen was suppressed by the blocking of collagen-integrin interaction. We could conclude that the effect of type I collagen was mediated via binding of collagen to integrin receptors. PMID- 12647293 TI - Regulation of hematopoietic-specific G-protein Galpha15 and Galpha16 by protein kinase C. AB - Heterotrimeric G proteins mediate cell growth and differentiation by coupling cell surface receptors to intracellular effector enzymes. The G-protein alpha subunit, Galpha(16), and its murine homologue Galpha(15), are expressed specifically in hematopoietic cells and their expression is highly regulated during differentiation of normal and leukemic cells. In this study, we examined the phosphorylation of Galpha(15)/Galpha(16) and its role in receptor and effector coupling. We observed a PMA-stimulated intact cell phosphorylation of Galpha(15) in COS7 cells transfected with Galpha(15) and protein kinase Calpha (PKCalpha), and phosphorylation of endogenous Galpha(16) in HL60 cells. We also showed that peptides derived from the two G-proteins were phosphorylated in vitro using purified brain PKC. Furthermore, we identified the putative phosphorylation site and showed that mutation or deletion of this PKC phosphorylation site inhibited phospholipase C (PLC) activation. The behavior of double mutants with the constitutively active G-protein mutation (QL-mutant) and mutation in the putative phosphorylation site suggests that the phosphorylation site of Galpha(15/16) is essential for receptor-coupled activation of PLC, but not for direct interaction of the G-protein with PLC-beta. PMID- 12647295 TI - Ref-1 protein enhances the IL-2-stimulated telomerase activity. AB - Telomerase is an important ribonucleoprotein enzyme involved in cellular proliferation and senescence. Activation of telomerase has been detected in a vast majority of human cancer cells. In this article, we demonstrated that Interleukin-2 (IL-2) which is the pivotal cytokine in the immune system could stimulate the activity of telomerase in the cultured BA/F3beta cells. It was also found that the level of IL-2-induced telomerase activity was decreased by the treatment with chemical oxidant in vitro. Since IL-2 stimulation produces a oxidative shift of the intracellular environment, the activation and maintenance of telomerase in this oxidative circumstance requires particular protection. Here we proved the redox factor-1 (Ref-1) protein was involved in this process. The addition of GST-Ref-1 protein increased the level of IL-2-induced telomerase activity in the TRAP assay, while elimination of the endogenous Ref-1 protein by immunodepletion decreased it. Consistent with these in vitro results, IL-2 induced telomerase activity could be enhanced by transient overexpression of Ref 1 protein in BA/F3beta cells. Taken together, these findings proved that Ref-1 protein benefits the activation of telomerase activity in the oxidative microenvironment of the BA/F3beta cells stimulated by IL-2. PMID- 12647297 TI - Regulation of bone resorption and osteoclast survival by nitric oxide: possible involvement of NMDA-receptor. AB - Nitric oxide has been shown to play an important role in regulation of bone resorption. However, the role of endogenous nitric oxide on osteoclast activity remains still controversial. In this work, using RT-PCR amplification, we demonstrated that rabbit mature osteoclasts express mRNA encoding for neuronal nitric oxide synthase suggesting that this enzyme could be involved in basal nitric oxide production in these cells. Then we assessed the effect of carboxy PTIO, a nitric oxide scavenger, on in vitro bone resorption and osteoclast survival. Carboxy-PTIO (10-100 microM) inhibited osteoclastic bone resorption in a dose dependent manner and induced osteoclast apoptosis by a mechanism involving caspase 3 activation. These results suggest that basal concentration of endogenous nitric oxide may be essential for normal bone resorption by supporting osteoclast survival. Because osteoclasts express N-methyl-d-aspartate-receptor (NMDA-R), we hypothesized that in osteoclasts NMDA-R may be involved in nitric oxide production as in neuronal cells. We confirmed that blockade of NMDA-R with specific non-competitive antagonists, MK801 and DEP, strongly inhibited bone resorption. As for carboxy-PTIO, we showed that blockade of NMDA-R by both antagonists induced osteoclast apoptosis in a dose dependent manner by a mechanism dependent on caspase 3 activation. Intracellular calcium concentration in osteoclasts decreased within minutes in the presence of both antagonists. Finally, MK801-induced osteoclast apoptosis was partially reversed in the presence of small amount of SNAP (100 nM), a nitric oxide donor, suggesting that the effect of NMDA-R on osteoclast apoptotic cell death could be due to a decrease in nitric oxide production. Taken together, our results are consistent with the hypothesis that NMDA-R on osteoclasts could have a similar function as those in neuronal cells, i.e., to allow a calcium influx, which in turn activates a constitutive neuronal nitric oxide synthase. Nitric oxide generated by this pathway may be essential for osteoclast survival and hence for normal bone resorption. PMID- 12647296 TI - Progression of chondrogenesis in C3H10T1/2 cells is associated with prolonged and tight regulation of ERK1/2. AB - Close contact of mesenchymal cells in vivo and also in super dense micromass cultures in vitro results in cellular condensation and alteration of existing cellular signaling required for initiation and progression of chondrogenesis. To investigate chondrogenesis related changes in the activity of ubiquitous cell signaling mediated by mitogen-activated protein kinases (MAP kinase), we have compared the effect of cell seeding of pluripotent C3H10T1/2 mesenchymal cells as monolayers (non-chondrogenic culture) or high density micromass cultures (chondrogenic) on the regulation and phosphorylation state of extracellular signal-regulated kinase 1 and 2 (ERK1/2) and also on regulation of ERK1/2 nuclear targets, namely, activation protein-1 (AP-1) and serum response factor (SRF). Increasing cell density resulted in reduced DNA binding as well as activity of AP 1. SRF activity, on the other hand, was up-regulated in confluent monolayer cultures but like AP-1 was inhibited in micromass cultures. Low levels of PD 98059 (5 microM), a specific inhibitor of ERK1/2, resulted in delayed induction of AP-1 and SRF activity whereas higher concentrations of this inhibitor (10-50 microM) conferred an opposite effect. Increasing concentrations of the PD 98059 inhibitor in long term monolayer or micromass cultures (2.5 day) resulted in differential regulation of c-Fos and c-Jun protein levels as well as total expression and phosphorylation levels of ERK1/2. PD 98059 treatment of C3H10T1/2 micromass cultures also resulted in up-regulation of type IIB collagen and Sox9 gene expression. While high expression of aggrecan and type IIB collagen genes were dependent on BMP-2 signaling, ERK inhibition of BMP-2 treated micromass cultures resulted in reduced activity of both genes. Our findings show that the activity of ERK1/2 in chondrogenic cultures of C3H10T1/2 cells is tightly controlled and can cross interact with other signaling activities mediated by BMP 2 to positively regulate chondrogensis. PMID- 12647298 TI - Characterization of PTH/PTHrP receptor in rat duodenum: effects of ageing. AB - In rat enterocytes, signaling through the parathyroid hormone (PTH)/PTH-related peptide receptor type 1(PTHR1) includes stimulation of adenylyl cyclase, increases of intracellular calcium, activation of phospholipase C, and the MAP kinase pathway, mechanisms that suffer alterations with ageing. The purpose of this study was to evaluate whether an alteration at the level of the PTH receptor (PTHR1) is the basis for impaired PTH signaling in aged rat enterocytes. Western Blot analysis with a specific monoclonal anti-PTHR1 antibody revealed that a 85 kDa PTH binding component, the size expected for the mature PTH/PTHrP receptor, localizes in the basolateral (BLM) and brush border (BBM) membranes of the enterocyte, being the protein expression about 7-fold higher in the BLM. Two other bands of 105 kDa (corresponding to highly glycosylated, incompletely processed receptor form) and 65 kDa (proteolytic fragment) were also seen. BLM PTHR1 protein expression significantly decreases with ageing, while no substantial decrease was observed in the BBM from old rats. PTHR1 immunoreactivity was also present in the nucleus where PTHR1 protein levels were similar in enterocytes from young and aged rats. Immunohistochemical analysis of rat duodenal sections showed localization of PTHR1 in epithelial cells all along the villus with intense staining of BBM, BLM, and cytoplasm. The nuclei of these cells were reactive to the PTHR1 antiserum, but not all cells showed the same nuclear staining. The receptor was also detected in the mucosae lamina propria cells, but was absent in globets cells from epithelia. In aged rats, PTHR1 immunoreactivity was diffused in both membranes and cytoplasm and again, PTH receptor expression was lower than in young animals, while the cell nuclei showed a similar staining pattern than in young rats. Ligand binding to PTHR1 was performed in purified BLM. rPTH(1-34) displaced [I(125)]PTH(1-34) binding to PTHR1 in a concentration-dependent fashion. In both, aged (24 months) and young (3 months) rats, binding of [I(125)]PTH was characterized by a single class of high-affinity binding sites. The affinity of the receptor for PTH was not affected by age. The maximum number of specific PTHR1 binding sites was decreased by 30% in old animals. The results of this study suggest that age-related declines in PTH regulation of signal transduction pathways in rat enterocytes may be due, in part, to the loss of hormone receptors. PMID- 12647299 TI - Stage specific inhibition of osteoblast lineage differentiation by FGF2 and noggin. AB - Fibroblast growth factor 2 (FGF2) and noggin are two unrelated ligands of two distinctly different signaling pathways that have a similar inhibitory effect on osteoblast differentiation. Because of their differences, we postulated that they probably acted at a different stage within the osteoprogenitor differentiation pathway. This study was performed on primary murine bone cell cultures under conditions where alkaline phosphatase (AP) and type I collagen expression (Col1a1) were observed by day 7 (preosteoblast stage), followed by bone syaloprotein (BSP) at day 11 (early osteoblast) and osteocalcin (OC) by day 15-18 (mature osteoblast stage). FGF2 completely inhibited expression of AP and the mRNA transcript for Col1a1, while noggin showed only a partial inhibition of these markers of preosteoblast differentiation. However, the markers of differentiated osteoblasts (BSP and OC) were completely inhibited in both the FGF2 and noggin treated cultures, suggesting that noggin acts at later point in the osteoprogenitor differentiation pathway than FGF2. To further verify that the inhibition was occurring at a different stage of osteoblasts development, primary cultures derived from transgenic mice harboring segments of the collagen promoter driving green fluorescent protein (GFP) that activate at different levels of osteoblast differentiation were analyzed. Consistent with the endogenous markers, pOBCol3.6GFP and pOBCOL2.3GFP transgene activity was completely inhibited by continuous addition of FGF2, while noggin showed partial inhibition of pOBCol3.6GFP and complete inhibition of the pOBCol2.3GFP transgene. Upon removal of either agent, endogenous and GFP markers of osteoblast differentiation reappeared although at a different temporal pattern. This work demonstrates that FGF2 and noggin can reversibly modulate osteoblast lineage differentiation at different maturational stages. These agents may be useful to enrich for and maintain a population of osteoprogenitor cells at a defined stage of differentiation. PMID- 12647300 TI - TE2 and TE1 sub-elements of the testis-specific histone H1t promoter are functionally different. AB - The testis-specific linker histone H1t gene is transcribed exclusively in pachytene primary spermatocytes. Tissue specific expression of the gene is mediated in part by transcriptional factors that bind elements located within the proximal and distal promoter. A 40 bp promoter element, designated H1t/TE, that is located within the proximal promoter between the CCAAT-box and AC-box, is known to be essential for H1t gene transcription in transgenic animals. In the present study, we show by SDS-PAGE analysis of UV crosslinked protein and DNA and by electrophoretic mobility shift assays (EMSA) of testis nuclear proteins separated on a non-denaturing glycerol gradient that the TE1 sub-element is bound by a protein complex. Mutation of TE1 leads to a drop in H1t promoter activity in germinal GC-2spd cells as well as in nongerminal Leydig, NIH3T3, and C127I cell lines. Although TE1 and TE2 sub-elements have similar sequences, mutation of the TE2 sub-element causes an increase in promoter activity in C127I and Leydig cells. The rat TE1 but not TE2 contains a CpG dinucleotide and this cytosine is methylated in liver but not in primary spermatocytes. Methylation of the cytosine at this site almost eliminates nuclear protein binding. Thus, there are significant functional differences in the TE2 and TE1 sub-elements of the H1t promoter with TE1 serving as a transcriptional activator binding site and TE2 serving as a repressor binding site in some cell lines. PMID- 12647301 TI - Vascular smooth muscle cells efficiently activate a new proteinase cascade involving plasminogen and fibronectin. AB - The plasminogen/plasmin system is involved in vascular wall remodeling after injury, through extracellular matrix (ECM) degradation and proteinase activation. Vascular smooth muscle cells (VSMCs) synthesize various components of the plasminogen/plasmin system. We investigated the conversion of plasminogen into plasmin in primary cultured rat VSMCs. VSMCs efficiently converted exogenous plasminogen into plasmin in a time- and dose-dependent manner. We measured plasmin activity by monitoring the hydrolysis of Tosyl-G-P-R-Mca, a fluorogenic substrate of plasmin. Cell-mediated plasmin activation was associated with the degradation of ECM, as revealed by fibronectin proteolysis. Plasmin also activated a proteinase able to hydrolyze Mca-P-L-G-L-Dpa-A-R-NH(2), a fluorogenic substrate of matrix metalloproteinases (MMPs). However, this proteinase was not inhibited by an MMP inhibitor. Furthermore, this proteinase displayed similar biochemical and pharmacological properties to fibronectin-proteinase, a recently identified zinc-dependent metalloproteinase located in the gelatin-binding domain of fibronectin. These results show that VSMCs convert exogenous plasminogen into plasmin in their pericellular environment. By hydrolyzing matrix protein plasmin activates a latent metalloproteinase that differs from MMP, fibronectin proteinase. This metalloproteinase may participate to vascular wall remodeling, in concert with other proteinases. PMID- 12647302 TI - A synthetic peptide of link protein stimulates the biosynthesis of collagens II, IX and proteoglycan by cells of the intervertebral disc. AB - To date, there have been no reports on the effect on disc cells of the intervertebral disc (IVD) of the amino terminal peptide of link protein (DHLSDNYTLDHDRAIH) (link N) which is generated by the cleavage of human link protein by stromelysins 1 and 2, gelatinase A and B, and collagenase between His(16) and Ile(17). However, link N has been shown to act as a growth factor and stimulate synthesis of proteoglycans and collagen by chondrocytes of human articular cartilage. There are also no studies on the effect of link N on type IX collagen in any tissue. In the studies reported here, a serum-free pellet culture system has been used to examine whether link N can play a role in maintaining the integrity of disc matrix, specifically at the level of matrix assembly by cells of the IVD. Using this culture system, we determined the capacity of link N to stimulate accumulation of these matrix proteins in the annulus fibrosus (AF) and nucleus pulposus (NP). Gross inspection of separate AF and NP pellet cultures in the absence of link N revealed a progressive increase in size and a transition from "spherical" to "polygonal" pellets after centrifugation. Addition of 10 ng/ml link N resulted in increased pellet sizes for both AF and NP pellet cultures. Link N increased proteoglycan, type II and type IX collagen contents with an increase in DNA content over time. This study demonstrates that link N can act directly on disc cells to stimulate matrix production, which involves increased accumulation of proteoglycan, and types II and IX collagens. This study also identifies the value of pellet cultures for studies of the IVD cells in a serum-free chemically defined medium, in which pellets can continue growing in size in response to growth factors with minimal cell loss. Link N may have value in stimulating the growth and regeneration of the damaged IVD. PMID- 12647303 TI - Nuclear localization of basic fibroblast growth factor is mediated by heparan sulfate proteoglycans through protein kinase C signaling. AB - Understanding the process of wound healing will provide valuable insight for the development of new strategies to treat diseases associated with improper regeneration, such as blindness induced by corneal scarring. Heparan sulfate proteoglycans (HSPG) are not normally expressed in the corneal stroma, but their presence at sites of injury suggests their involvement in the wound healing response. Primary cultured corneal stromal fibroblasts constitutively express HSPG and represent an injured phenotype. Recently, nuclear localization of HSPG was shown to increase in corneal stromal fibroblasts plated on fibronectin (FN), an extracellular matrix protein whose appearance in the corneal stroma correlates with injury. One possible role for the nuclear localization of HSPG is to function as a shuttle for the nuclear transport of heparin-binding growth factors, such as basic fibroblast growth factor (FGF-2). Once in the nucleus, these growth factors might directly modulate cellular activities. To investigate this hypothesis, cells were treated with (125)I-labelled FGF-2 under various conditions and fractionated. Our results show that nuclear localization of FGF-2 was increased in cells plated on FN compared to those on collagen type I (CO). Interestingly, FGF-2-stimulated proliferation was increased in cells plated on FN compared to CO and this effect was absent in the presence of heparinase III. Furthermore, pre-treatment with heparinase III decreased nuclear FGF-2, and CHO cells defective in the ability to properly synthesize heparan sulfate chains showed reduced nuclear FGF-2 indicating that the heparan sulfate chains of HSPG are critical for this process. HSPG signaling, particularly through the cytoplasmic tails of syndecans, was investigated as a potential mechanism for the nuclear localization of FGF-2. Treatment with phorbol 12-myristate-13-acetate (PMA), under conditions that caused downregulation of protein kinase Calpha (PKCalpha), decreased nuclear FGF-2. Using pharmacological inhibitors of specific PKC isozymes, we elucidated a potential mode of regulation whereby PKCalpha mediates the nuclear localization of FGF-2 and PKCdelta inhibits it. Our studies suggest a novel mechanism in which FGF-2 translocates to the nucleus in response to injury. PMID- 12647304 TI - Suppressive role of endogenous regucalcin in the enhancement of nitric oxide synthase activity in liver cytosol of normal and regucalcin transgenic rats. AB - The suppressive role of endogenous regucalcin, which is a regulatory protein of calcium signaling, in the enhancement of nitric oxide (NO) synthase activity in the liver cytosol of rats was investigated. The enzyme activity was measured in a reaction mixture containing either vehicle or calcium chloride (1-20 microM) in the absence or presence of regucalcin (0.1, 0.25, or 0.5 microM). NO synthase activity was significantly increased by the addition of calcium (5-20 microM). This increase was completely abolished in the presence of trifluoperazine (TFP; 10-50 microM), an antagonist of Ca(2+)/calmodulin. The addition of regucalcin (0.1-0.5 microM) caused a significant fall in the calcium-increased enzyme activity. The effect of regucalcin (0.25 microM) in decreasing NO synthase activity was seen in the presence of ethylene glycol bis-(2-aminoethylether) N,N,N',N'-tetraacetic acid (EGTA, 1 mM) or TFP (20 microM), indicating that regucalcin acts independent on Ca(2+)/calmodulin. NO synthase activity was significantly raised in the presence of anti-regucalcin monoclonal antibody (10 50 ng/ml) in the reaction mixture. The effect of the antibody (50 ng/ml) or calcium (10 microM) in elevating NO synthase activity in the liver cytosol of normal rats was not seen in the liver cytosol obtained from regucalcin transgenic rats. Moreover, the increase in NO synthase activity in the liver cytosol of normal rats induced by a single intraperitoneal administration of calcium (5.0 mg/100 g body weight) was significantly enhanced in the presence of anti regucalcin monoclonal antibody (50 ng/ml) in the reaction mixture. The administration of calcium caused a significant increase in regucalcin level in the liver cytosol of normal rats. The present study demonstrated that endogenous regucalcin plays a suppressive role in the enhancement of NO synthase activity in the liver cytosol of rats. PMID- 12647305 TI - Involvement of MEKK1/ERK/P21Waf1/Cip1 signal transduction pathway in inhibition of IGF-I-mediated cell growth response by methylglyoxal. AB - The abnormal accumulation of methylglyoxal (MG), a physiological glucose metabolite, is strongly related to the development of diabetic complications by affecting the metabolism and functions of organs and tissues. These disturbances could modify the cell response to hormones and growth factors, including insulin like growth factor-1 (IGF-I). In this study, we investigated the effect of MG on IGF-I-induced cell proliferation and the mechanism of the effect in two cell lines, a human embryonic kidney cell line (HEK293), and a mouse fibroblast cell line (NIH3T3). MG rendered these cells resistant to the mitogenic action of IGF I, and this was associated with stronger and prolonged activation of ERK and over expression of P21(Waf1/Cip1). The synergistic effect of MG with IGF-I in activation of ERK was completely abolished by PD98059 but not by a specific PI3K inhibitor, LY294002, or a specific PKC inhibitor, bisindolylmaleimide. Blocking of Raf-1 activity by expression of a dominant negative form of Raf-1 did not reduce the enhancing effect of MG on IGF-I-induced activation of ERK. However, transfection of a catalytically inactive form of MEKK1 resulted in inactivation of the MG-induced activation of ERK and partial inhibition of the enhanced activation of ERK and over-expression of p21(Waf1/Cip1) induced by co-stimulation of MG and IGF-I. These results suggested that the alteration of intracellular milieu induced by MG through a MEKK1-mediated and PI3K/PKC/Raf-1-independent pathway resulted in the modification of cell response to IGF-I for p21(Waf1/Cip1) mediated growth arrest, which may be one of the crucial mechanisms for MG to promote the development of chronic clinical complications in diabetes. PMID- 12647306 TI - Inhibition of phosphatidylinositol 3-kinase and p70S6 kinase blocks osteogenic protein-1 induction of alkaline phosphatase activity in fetal rat calvaria cells. AB - Published studies reveal that Osteogenic Protein-1 (OP-1) and insulin-like growth factor-I (IGF-I) synergistically stimulate alkaline phosphatase (AP) activity and bone nodule formation in fetal rat calvaria (FRC) cells. In the present study, we examined whether there are interactions between the signal transduction pathways activated by these two growth factors. OP-1 did not significantly affect the levels of IRS-1, IRS-2, the p85alpha subunit of phosphatidylinositol 3-kinase (PI 3-kinase) or the extracellular signal-regulated kinase (ERK)-2, but stimulated ERK-1 protein by twofold. OP-1 also induced phosphorylation of ERK-1 and -2, but not of Akt/protein kinase B (PKB), a protein kinase that is downstream of PI 3 kinase. By comparison, IGF-I increased the levels of the phosphorylated forms of ERK-1 and -2, and Akt/PKB. Inhibition of ERK activation by PD98059 did not significantly alter the stimulation of AP activity by OP-1 or OP-1 in combination with IGF-I. In contrast, inhibition of PI 3-kinase activity by LY294002 blocked the induction of AP activity by OP-1 and OP-1 plus IGF-I. Treatment of cells with rapamycin, an inhibitor of the mammalian target of mTOR, resulted in a 47% and a 53% decrease in the AP activity induced by OP-1 alone and by OP-1 plus IGF-I, respectively. These studies suggest that PI 3-kinase and mTOR contribute to the induction of AP activity by OP-1 and the synergistic effect of OP-1 and IGF-I on AP activity in FRC cells. PMID- 12647307 TI - Effects of Bafilomycin A1: an inhibitor of vacuolar H (+)-ATPases on endocytosis and apoptosis in RAW cells and RAW cell-derived osteoclasts. AB - Bafilomycin A1, a specific inhibitor of V-ATPases, is a potent inhibitor of bone resorption, but the underlying mechanisms of its action remain unclear. In this study, we investigated the effect of Bafilomycin A1 on endocytosis and apoptosis in RAW cells and RAW cell-derived osteoclasts. Quantitative analysis by flow cytometry showed that Bafilomycin A1 increased total transferrin levels when RAW cells were exposed to labeled transferrin and decreased the total uptake of Dextran-rhodamine B, both in a dose- and time-dependent fashion, indicating that Bafilomycin influences receptor-mediated and fluid phase endocytosis in these cells. Furthermore, Bafilomycin A1 induced apoptosis of RAW cells in a dose dependent manner as evidenced by Annexin V flow cytometry. The action of Bafilomycin A1 on endocytotic events appeared to be more sensitive and occurred earlier than on its apoptosis inducing effects, suggesting that interrupting of endocytosis might be an early sign of Bafilomycin-mediated osteoclast inhibition. Semi-quantitative RT-PCR analysis showed that the gene transcripts of putative Bafilomycin A1 binding subunit, V-ATPase-subunit a3, were expressed in the preosteoclastic RAW cell line, and up-regulated during RANKL-induced osteoclastogenesis. Osteoclasts treated with Bafilomycin A1 exhibited apoptosis as well as altered cellular localization of Transferrin Alexa 647. Given that endocytosis and apoptosis are important processes during osteoclastic bone resorption, the potent effect of Bafilomycin A1 on endocytosis and apoptosis of osteoclasts and their precursor cells may in part account for Bafilomycin A1 inhibited bone resorption. PMID- 12647308 TI - Capacitative calcium influx in human epithelial breast cancer and non-tumorigenic cells occurs through Ca2+ entry pathways with different permeabilities to divalent cations. AB - The operation of capacitative Ca(2+) entry (CCE) in human breast cancer (SKBR3) and non-tumorigenic (HBL100) cell lines was investigated as an alternative Ca(2+) entry route in these cells. Ca(2+) readdition after thapsigargin-induced store depletion showed activation of CCE in both cell lines. SKBR3 cells exhibited retarded store depletion and CCE decay kinetics compared to the non-tumorigenic HBL100 cells, suggesting alterations in Ca(2+) homeostasis. CCE was also highly permeable to Mn(2+) and to a lesser extent to Sr(2+), but not to Ba(2+). In HBL100 cells, CCE is contributed (30%) by a Ca(2+)/Mn(2+) permeable route insensitive to low (1 microM) Gd(3+) and a Ca(2+)/Sr(2+)/Mn(2+) permeable non selective pathway (70%) sensitive to 1 microM Gd(3+). In SKBR3 cells, the relative contribution to CCE of both routes was opposite to that in non tumorigenic cells. PMID- 12647309 TI - Nuclear trafficking of FGFR1: a role for the transmembrane domain. AB - Several members of the fibroblast growth factor (FGF) family lack signal peptide (SP) sequences and are present only in trace amounts outside the cell. However, these proteins contain nuclear localization signals (NLS) and accumulate in the cell nucleus. Our studies have shown that full length FGF receptor 1 (FGFR1) accumulates within the nuclear interior in parallel with FGF-2. We tested the hypothesis that an atypical transmembrane domain (TM) plays a role in FGFR1 trafficking into the nuclear interior. With FGFR1 destined for constitutive fusion with the plasma membrane due to its SP, how the receptor may enter the nucleus is unclear. Sequence analysis identified that FGFR1 has an atypical TM containing short stretches of hydrophobic amino acids (a.a.) interrupted by polar a.a. The beta-sheet is the predicted conformation of the FGFR1 TM, in contrast to the alpha-helical conformation of other single TM tyrosine kinase receptors, including FGFR4. Receptor trafficking in live cells was studied by confocal microscopy via C-terminal FGFR1 fusions to enhanced green fluorescent protein (EGFP) and confirmed by subcellular fractionation and Western immunoblotting. Nuclear entry of FGFR1-EGFP was independent of karyokinessis, and was observed in rapidly proliferating human TE671 cells, in slower proliferating glioma SF763 and post-mitotic bovine adrenal medullary cells (BAMC). In contrast, a chimeric FGFR1/R4-EGFP, where the TM of FGFR1 was replaced with that of FGFR4, was associated with membranes (golgi-ER, plasma, and nuclear), but was absent from the nucleus and cytosol. FGFR1delta-EGFP mutants, with hydrophobic TM a.a. replaced with polar a.a., showed reduced association with membranes and increased cytosolic/nuclear accumulation with an increase in TM hydrophilicity. FGFR1(TM-) EGFP (TM deleted), was detected in the golgi-ER vesicles, cytosol, and nuclear interior; thus demonstrating that the FGFR1 TM does not function as a NLS. To test whether cytosolic FGFR1 provides a source of nuclear FGFR1, cells were transfected with FGFR1(SP-) (SP was deleted), resulting in cytosolic, non membrane, protein accumulation in the cytosol and the cell nucleus. Our results indicate that an unstable association with cellular membranes is responsible for the release of FGFR1 into the cytosol and cytosolic FGFR1 constitutes the source of the nuclear receptor. PMID- 12647310 TI - Highly potent and selective inhibition of varicella-zoster virus replication by bicyclic furo[2,3-d]pyrimidine nucleoside analogues. AB - The bicyclic furo[2,3-d]pyrimidine nucleoside analogues represent an entirely new class of fused furopyrimidine derivatives with unprecedented selectivity for varicella-zoster virus (VZV). From extensive structure-activity relationship (SAR) studies, the 6-(p-alkylphenyl)substituted furopyrimidine derivatives Cf 1742 and Cf 1743 emerged as the most potent inhibitors of VZV replication: they were found to inhibit both laboratory VZV strains and clinical VZV isolates at subnanomolar concentrations, while not being toxic to the host cells at 100,000 fold higher concentrations. Although the precise mechanism of action of these compounds remains to be elucidated, it is clear that for their antiviral activity they depend on phosphorylation by the VZV-encoded thymidine kinase. The furo[2,3 d]pyrimidine nucleoside analogues are not susceptible to degradation by human or bacterial thymidine phosphorylase, which may otherwise release the free aglycone. Also, the latter is not inhibitory to dihydropyrimidine dehydrogenase, an enzyme involved in the degradation of thymine, uracil, and the anticancer agent 5 fluorouracil. Further development of the furo[2,3-d]pyrimidine nucleoside analogues as new therapeutic modalities for the treatment of VZV infections (i.e., varicella and herpes zoster) seems highly justified. PMID- 12647311 TI - In silico prediction of drug-binding strengths to human serum albumin. AB - Drug binding to Human Serum Albumin (HSA) is an area of intense research. The pharmacokinetics and pharmacodynamics of drugs are strongly affected by their binding to this protein. In this article, the field is reviewed, as well as our models to predict drug-binding affinities to HSA from drug structure. The physiological role of HSA is described, as well as its influence in drug action. The crystal structures of this protein are discussed, emphasizing the two drug binding sites and the fatty acids binding sites observed therein. The advantages of using high-performance affinity chromatography to rapidly screen drugs for HSA binding are explained. The different QSAR models for HSA binding of restricted families of drugs (both from other groups and our group) are enumerated. Finally, a detailed description of our general models to predict drug-binding strengths to HSA from structure is given. It is expected for these models to be useful in drug design and pharmaceutical research. PMID- 12647312 TI - Selection criteria for drug-like compounds. AB - The fast identification of quality lead compounds in the pharmaceutical industry through a combination of high throughput synthesis and screening has become more challenging in recent years. Although the number of available compounds for high throughput screening (HTS) has dramatically increased, large-scale random combinatorial libraries have contributed proportionally less to identify novel leads for drug discovery projects. Therefore, the concept of 'drug-likeness' of compound selections has become a focus in recent years. In parallel, the low success rate of converting lead compounds into drugs often due to unfavorable pharmacokinetic parameters has sparked a renewed interest in understanding more clearly what makes a compound drug-like. Various approaches have been devised to address the drug-likeness of molecules employing retrospective analyses of known drug collections as well as attempting to capture 'chemical wisdom' in algorithms. For example, simple property counting schemes, machine learning methods, regression models, and clustering methods have been employed to distinguish between drugs and non-drugs. Here we review computational techniques to address the drug-likeness of compound selections and offer an outlook for the further development of the field. PMID- 12647313 TI - Recent progress in the development of coumarin derivatives as potent anti-HIV agents. AB - Numerous plant-derived compounds have been evaluated for inhibitory effects against HIV replication, and some coumarins have been found to inhibit different stages in the HIV replication cycle. This review article describes recent progress in the discovery, structure modification, and structure-activity relationship studies of potent anti-HIV coumarin derivatives. A dicamphanoyl khellactone (DCK) analog, which was discovered and developed in our laboratory, and calanolide A are currently in preclinical studies and clinical trials, respectively. PMID- 12647314 TI - Boronic acid compounds as potential pharmaceutical agents. AB - Boronic acid compounds have been used, because of their unique structural features, for the development of potent enzyme inhibitors, boron neutron capture agents for cancer therapy, and as antibody mimics that recognize biologically important saccharides. Consequently, there has been a surge of interests in boronic acid compounds. This study reviews the recent development in this area during the last six years. PMID- 12647315 TI - VLA-4 antagonists: potent inhibitors of lymphocyte migration. AB - Circulating lymphocytes normally migrate through extravascular spaces in relatively low numbers as important members of the immunosurveillance process. That is until signals are received by endothelial cells that there is an underlying infection or inflammatory condition. These vascular surface cells in turn overexpress and present ligands to circulating lymphocyte adhesion molecules. Upon encountering this higher density of ligands, lymphocytes, which had been leisurely rolling along the vascular surface, now become more firmly attached, change shape, and migrate through tight junctions to the sites of infection or inflammation. If the initiating events are not resolved and the condition becomes chronic, there can be a sustained extravasation of lymphocytes that can exacerbate the inflammatory condition, which in turn will continue to recruit more inflammatory cells resulting in unwanted tissue destruction. It is for the attenuation of this cycle of sustained inflammatory cell recruitment that very late activating antigen-4 (VLA-4) antagonists are being developed. Most lymphocytes, except neutrophils, express VLA-4 on their surface and they interact with endothelial vascular cell adhesion molecule-1 (VCAM-1). It is this interaction that VLA-4 antagonists are intended to disrupt, thus, putting an end to the cycle of chronic inflammation, which is the hallmark of many diseases. This review will provide an update of VLA-4 antagonists that have appeared since early 2001 and will discuss some of the issues, both positive and negative, that may be encountered in their development. PMID- 12647316 TI - [A clinical analysis of alcoholic liver cirrhosis in South Sichuan area]. PMID- 12647317 TI - [Three-dimensional reconstruction and measurement of the human membranous semicircular canal]. AB - The planar relationship of the human semicircular canals was determined by Blanks et al. at a series of points measured from the dissected bony labyrinth of the human skull. The relationship of membranous canal planes have not, however, been measured from the human temporal bone. We reconstructed 3 semicircular canals by computer-aided 3-dimensional analysis and measured the angles formed between pairs of 3 osseous and membranous canal planes of temporal bones. Five temporal bones in adults were used for this study. Results indicated angles formed between pairs of ipsi-lateral canal planes of both the bony and membranous labyrinth. Angles formed between the horizontal-anterior, anterior-posterior, and posterior horizontal canal planes of the bony labyrinth were 89.64 +/- 1.82 (mean +/- SD), 90.95 +/- 1.25, and 94.02 +/- 3.77 degrees. The same angles measured from the membranous labyrinth were 90.12 +/- 2.64, 90.18 +/- 2.75, and 91.48 +/- 6.32 degrees. Differences between the angles formed between bony and membranous canal planes were 2.11, 6.05, and 3.26 degrees in the anterior, horizontal, and posterior canal. Pairs of membranous canal planes were nearly perpendicular without exception, but pairs of osseous canal planes had a larger deviation from 90 degrees. This suggested that membranous canals could successfully be constructed in adequate alignment for canal function in the large perilymphatic space within osseous semicircular canals. PMID- 12647319 TI - [Impact of the "Doctor-Heli" system for emergency and critical care medicine in otolaryngology]. AB - We reported the "Doctor-Heli" (medical service helicopter) system at the center of emergency and critical care medicine at Tokai University Hospital. From October 1999 to March 2001, the service had transported 485 patients, shortening the time to critical care and improving patient-prognosis. We report a case of cervical and laryngeal trauma occurring during a suicide attempt successfully treated thanks to the rapid start of critical care enabled by use of the helicopter. The service has proven its utility in otolaryngology and head and neck surgery. PMID- 12647318 TI - [Head and neck cancer treatment in the elderly--evaluation and management of complications]. AB - With the population over age 70 growing, treatment for head and neck cancer in the elderly has increased. We retrospectively evaluated their management and outcome. Subjects numbered 121--83 men and 38 women from 70 to 94 years old- initially treated at our hospital. We classified them into 2 groups by age--the aged at 70-79 years (55 men and 26 women) and the very old at 80 years and older (28 men and 12 women). We also evaluated a younger control group aged 50-59 years (37 men and 19 women). Primary tumor sites were the oral cavity (28.1%), larynx (28.1%), paranasal sinus (15.8%), and hypopharynx (9.9%). Preoperative geriatric disease was seen in 54% of controls, 74% of the aged, and 93% of the very old. Cardiovascular and respiratory diseases were most common. Surgical treatment and irradiation were essential for cancer treatment. Postoperative complications, including pneumonia, delirium, renal and cardiovascular hypofunction occurred in 56.5% of controls, 48.2% of the aged, and 47.8% of the very old. The frequency of postoperative complications correlated significantly with the American Society of Anesthesiologist classification of physical status (ASA) and preoperative performance status (PS). The complications of irradiation including pneumonia, dehydration, and feeding disturbance occurred in 53% of the very old. Cures were achieved in 83.9% of controls, 81.5% of the aged, and 65.0% of the very old. Cause specific 5-year survival in those cured was 85.2% of controls, 84.5% of the aged, and 80.0% of the very old. Median survival in those not cured was 4 months in controls, 9.6 months in the aged, and 5 months in the very old. We concluded that curative treatment is important in the elderly, and the success of curative treatment and the prevention of complications depend on careful assessment of systemic disease, PS, ASA, and mental activity. PMID- 12647320 TI - [Study on 242 inpatients reporting vertigo and dizziness]. AB - We studied 242 inpatients--men (34%) and women (66%) reporting vertigo and dizziness while hospitalized at Hakodate Municipal Hospital from July 1999 to June 2002. Adults over 65 years old accounted for 47.3% of all subjects. Reports of symptoms tended to increase in March, August, and December. Cases were classified into 4 groups: 1) peripheral disorders (35.7%), 2) central disorders (7.9%), 3) disorders of other origins (8.3%), and 4) disorders of unknown origin (47.1%). Group 1) involved vestibular neuritis (10.1% of all cases), Meniere diseases (12.4%), benign paroxysmal positional vertigo (BPPV) (5.8%), and sudden deafness with vertigo (5.8%). Patients with vestibular neuritis--25 subjects including men (68%) and women (32%)--tended to be admitted mainly in July. Vestibular compensation in 15 patients with vestibular neuritis was studied using the platform stabilometry. The total length of a locus and the area of surroundings with open eyes decreased significantly (p < 0.05) as the condition of patients improved. In conclusion, many subjects reporting vertigo and dizziness had vestibular neuritis and few had BPPV. We found platform stabilometry to be useful in estimation of improvement in patients with vestibular neuritis. PMID- 12647321 TI - [Effects of Burow's solution as an ear drop on Intractable chronic suppurative diseases of the external ear canal and middle ear]. AB - OBJECTIVE: Burow's solution was developed by Karl August von Burow in the 19th century as an ear drop. Thorp et al. reported its excellent effect on chronic suppurative otitis media in 1998. We applied Burow's solution to intractable chronic purulent diseases of the external and middle ear in the 12 months from February 2001. SUBJECTS: Subjects were 25 ears of 21 patients--35-79 years old, 10 men's ears and 15 women's ears--whose disease has continued for an average of 3.78 years. Diseases and patients are as follows: 1) 11 ears with postoperative mastoid cavity problems, 2) 7 ears with chronic external otitis and chronic eczema of the external ear canal, 3) 7 ears with fungal otitis externa, 4) 6 ears with chronic otitis media with perforation, and 5) 2 ears of chronic granulated myringitis. METHOD: The solution was dropped into the ear canal once a day for 10 min or cotton balls soaked with the solution were applied to the canal wall. Criteria of efficacy were divided into cured, effective and unchanged. RESULTS: 8/11 ears with a cavity problem were cured (72.7%) and 3 ears showed treatment to be effective. All 7 ears with chronic external otitis and chronic eczema were cured. All 7 ears with fungal otitis externa were cured. 4/6 ears (66.6%) with chronic perforated otitis media were cured and 1 ear each showed treatment to be effective and 1 unchanged. Two chronic granurated myringitis were cured. Twenty of 25 ears (80%) were cured. Pre- and posttreatment audiometries showed almost the same. The cure appeared within 3 days to 3 weeks. The solution was not effective against mucoid secretion, cholesteatoma, or residual mastoid cells. The day after treatment, serous secretion appears temporarily. CONCLUSION: Burows's solution is seemed to be very effective and nonototoxic as an otic drop for treating suppurative ear diseases. PMID- 12647322 TI - Mediational models of spirituality and depressive symptomatology among HIV positive Puerto Rican women. AB - A survey of 142 Puerto Rican women living with HIV/AIDS in New York City revealed high Centers for Epidemiological Studies Depression (CES-D) scores, with 66% of the sample scoring above the conventional threshold of possible clinical depression. Most respondents (71%) were Catholic, 29% considered themselves members of a church or other place of worship, and 30% reported attending religious services 1-3 times a month. As predicted, spirituality was high and negatively associated with CES-D scores. A series of simultaneous multiple regression analyses controlling for all potentially confounding medical and sociodemographic variables demonstrated that both mastery and self-esteem scores mediated this relationship. Implications for future research and the provision of services to HIV-positive Puerto Rican women are discussed. PMID- 12647323 TI - The measurement of self-esteem among Korean Americans: a cross-ethnic study. AB - Studies have indicated that Asians' self-esteem is generally lower than other ethnic groups'. The authors hypothesized that an item response bias would result in lower self-esteem scores for Asians. Specifically, it was hypothesized that positively worded items would be rated lower by Korean Americans but that there would be no difference on negatively worded items across ethnic groups. The hypothesis was tested using 214 individuals diagnosed with schizophrenia who were Korean American, Caucasian, African American, or Latino. Results supported the hypothesis. Korean Americans had significantly lower self-esteem scores than the other 3 groups on all positively worded items but were no different from the other groups on negatively worded items. Additional analyses showed modest support for the cross-cultural construct equivalence of this measure of self esteem. PMID- 12647324 TI - Age, acculturation, cultural adjustment, and mental health symptoms of Chinese, Korean, and Japanese immigrant youths. AB - This study of Chinese, Japanese, and Korean immigrant junior high and high school students (N = 319) investigated the association between age, acculturation, cultural adjustment difficulties, and general mental health concerns. Hierarchical regression analyses determined that among all of the independent variables, age, acculturation, and cultural adjustment difficulties had significant predictive effects on mental health symptoms. Implications for theory, research, and practice are addressed, particularly as they relate to developmental issues among immigrant youths. PMID- 12647325 TI - The effects of depression on the emotional responses of Spanish-speaking Latinas. AB - Emotional responses (physiology, self-report, and facial expression) of 12 depressed and 10 nondepressed Spanish-speaking Latinas during sad and amusing film clips of human and animal content were compared. Depressed Latinas demonstrated less electrodermal reactivity across all the film clips and displayed fewer social smiles during the amusing-human film clip than nondepressed Latinas. No differences emerged for cardiovascular measures, reports of emotion, or facial expressions of happiness and negative emotion. Observed differences in electrodermal reactivity are similar to results from previous studies of Anglo Americans, suggesting that reduced electrodermal activity may be linked to depression across cultures. The findings also suggest that, for Latinas, depression may selectively alter expressions that serve interpersonal functions. PMID- 12647326 TI - Perfectionism and ethnicity: implications for depressive symptoms and self reported academic achievement. AB - Ethnic differences in perfectionism were examined among Asian American, African American, and Caucasian American college students. Analyses revealed that Asian American students scored significantly higher than the other groups on 3 of 6 perfectionism subscales. Minority students reported significantly higher parental expectations than Caucasian students. Significant correlations emerged for all 3 groups between depressive symptoms and concerns about making mistakes, perceived criticism from parents, and self-doubt. In regression analyses, perfectionism explained significant variance in depressive symptoms for Asian Americans and Caucasians and explained significant variance in cumulative grade point average for Asian Americans and African Americans. Self-doubt emerged as a robust individual predictor in these analyses. PMID- 12647327 TI - Jewish identity and intentions to obtain breast cancer screening. AB - This study explored the associations between 2 dimensions of Jewish identity (cultural identification and religious practice) and intentions to perform breast cancer screening. Ashkenazi Jewish women (N = 220) completed surveys as part of an ongoing study of breast cancer risk counseling. Multiple regressions examined the relationships between the 2 identity measures and intention to follow routine recommendations for mammography, intention to perform monthly breast self-exam, and interest in genetic testing for breast cancer susceptibility. Cultural identity positively predicted interest in testing, whereas religious identity was inversely related. Religious identity was a significant predictor of intention to adhere to mammography recommendations. Findings show that culture and religion, although correlated, may have different associations with health attitudes. PMID- 12647329 TI - [Zoonosis and ocular diseases]. PMID- 12647328 TI - Examining culturally based variables associated with ethnicity: influences on credibility perceptions of empirically supported interventions. AB - Treatment rationales for 2 widely used and empirically supported interventions, cognitive therapy (CT) and time-limited dynamic psychotherapy (TLDP), were examined for their perceived credibility among 136 Asian American college students. This study conducted a comprehensive analysis of culturally based variables (often assumed to underlie ethnicity) and their related effects on credibility perceptions. Variables assessed included cultural identity, self construals, values, and mental health beliefs. Participants were randomly assigned to read either a CT or TLDP treatment rationale for depression and then rated the credibility of the interventions. Results indicated that cultural identity and self-construals moderated credibility ratings across CT and TLDP rationales. Findings underscore the importance of moving beyond ethnic group analyses to the examination of specific culturally based variables. PMID- 12647330 TI - [Long-term complications of MIRAgel exoplants in scleral buckling surgery]. AB - BACKGROUND: Long-term complications of extrusion have been reported in patients who received MIRAgel buckles for retinal detachment. All ophthalmologists need to recognize the clinical characteristics of these severe complications, and how the clinical course of complication after the use of MIRAgel buckles differs from that of silicone buckles. METHODS: Case reports and a review of the literature. RESULTS: Severe deterioration of MIRAgel buckles may occur when left in place for 5 years or longer, causing adverse symptoms and occasionally intraocular complications. This deterioration is associated with changes in the microstructural architecture of the hydrogel material in the MIRAgel buckles. CONCLUSION: Long-term follow-up is necessary for patients with MIRAgel buckles, and the buckles should be removed prophylactically when advisable. When severe ocular motility disturbances and/or presence of a tumor-like extraocular mass are observed in patients several years after retinal detachment surgery, the cause may be a swollen MIRAgel buckle. PMID- 12647331 TI - [Electrophysiological and histopathological study on visual toxicity of clioquinol]. AB - OBJECTIVE: To evaluate the effect of clioquinol on the optic nerve and retina of rhesus monkeys by ophthalmoscopy, electrophysiology and histopathology. METHODS: Clioquinol was given orally to 5 monkeys, gradually increasingly from 100 mg/kg/day up to 14 months(total dosage 227 g/kg). Ophthalmoscopy, erectroretinogram(ERG), visual evoked potential(VEP) and histopathological examination of enucleated eyeballs were done periodically up to 10 years. RESULTS: The margin of the optic disc was not clear at the early stage, but the colour became atrophic at the late stage. VEP maximum amplitude decreased quickly at the early stage and the amplitude of ERG a and b waves and oscillatory potential decreased gradually. 37 months after the discontinuation of administration of VEP, ERG amplitude increased gradually. Swelling of axons and disorganization of the myelin sheath were noticed 2.5 months after beginning treatment. Swelling of the peripapillary nerve fiber layer was seen 5.5 months after beginning treatment. Karyorrhexis was seen in the inner layer of the retina after 12.5 months. Axonal swelling disappeared and the myelin sheath became reorganized 9 months after the discontinuation of treatment. CONCLUSIONS: Clioquinol produced an early decrease of electrophysiological function, but recovery of function was seen after discontinuation of treatment. The degeneration of axons and myelin sheath continued during treatment, and interruption of the degeneration was seen after discontinuation of treatment. PMID- 12647332 TI - [Correlation of the presence of meibomian gland dysfunction with the incidence of superficial punctate keratopathy after penetrating keratoplasty]. AB - PURPOSE: To examine the correlation of the presence of meibomian gland dysfunction(MGD) with the incidence of superficial punctate keratopathy (SPK) after penetrating keratoplasty. SUBJECTS & METHODS: We studied 151 eyes of 141 patients that underwent penetrating keratoplasty. SPK and the presence of MGD were evaluated by slit-lamp examination. Tear function was evaluated by Schirmer test. RESULTS: Among 151 eyes investigated, 78 eyes had neither MGD nor hypolacrimation. The number of eyes with MGD only, with hypolacrimation only, or with both MGD and hypolacrimation was 34, 23, and 16. The presence of SPK after penetrating keratoplasty was correlated with the presence of MGD 1 month or 6 months post-surgery, but no correlation was observed with the presence of hypolacrimation. There was no interaction between the presence of MGD and hypolacrimation and the incidence of SPK. CONCLUSION: This study suggested that the active treatment of MGD is important in the post-surgery management of SPK. PMID- 12647334 TI - [Long-term clinical course of excimer laser photorefractive keratectomy]. AB - PURPOSE: Long-term clinical follow-up for 5 years or longer has not yet been reported in Japan. We performed excimer laser photorefractive keratectomy(PRK) on 181 eyes from 1990 to 1993, and 44 of those eyes could be followed for 8 to 10 years. METHODS: The 44 eyes were divided into 3 groups depending on degree of correction. We compared of safety, efficacy, predictability-spherical equivalent, stability, complication, and satisfaction among the 3 groups. RESULTS AND CONCLUSIONS: The first generation PRK was certified clinically to be safe and stable in patients corrected for low to moderate myopia. PMID- 12647333 TI - [Attitude survey of diabetic patients visiting the department of internal medicine as outpatients]. AB - PURPOSE: To investigate the awareness and understanding of ophthalmologic complications by diabetic patients and assess the problems. SUBJECTS AND METHODS: In August 1999, a questionnaire survey was done of 3,613 diabetic outpatients attending the Department of Internal Medicine at Tokyo Women's Medical University Diabetes Center. RESULTS: Patients who understood diabetic retinopathy as ophthalmological complications were 54.4% and those to whom physicians explained the complications were 33.8%. Patients who were recommended to receive an eye examination by physicians were 66.4% and those who continued to attend the eye clinic were 61.8%. Patients who had ophthalmological complications explained by physicians were 74.2%. CONCLUSION: Few patients understood diabetic retinopathy as a diabetic ophthalmological complication. To encourage the awareness and understanding concerning diabetic retinopathy, it is important to form a tight relationship between physicians and ophthalmologists and to improve the system for educating patients. PMID- 12647335 TI - [Cat scratch disease with posterior segment involvement]. AB - PURPOSE: To describe the clinical characteristics of patients with cat scratch disease during the last 2 years. METHODS: Clinical characteristics and anterior and posterior segment manifestations were reviewed in five patients who were serologically diagnosed as having cat scratch disease. RESULTS: Four women and one man were examined. Their ages ranged from 7 to 60 years. Each patient had a markedly elevated serum anti-Bartonella henselae antibody titer. Visual symptoms developed 2 weeks or less after the onset of systemic symptoms. Lymphadenopathy was detected in one of five patients. Neuroretinitis was found in 4 patients, and papillitis in 1 patient. Seven eyes showed retinochoroidal exudates. Anterior uveitis was observed in three eyes. Four patients received systemic corticosteroids. CONCLUSION: The ocular manifestations of cat scratch disease include neuroretinitis, papillitis, retinochoroidal exudates, and anterior uveitis. PMID- 12647336 TI - [A case of corneal lactoferrin amyloidosis secondary to trichiasis]. AB - PURPOSE: A case of corneal lactoferrin amyloidosis secondary to trichiasis is reported. CASE: A 30-year-old male suffered from trichiasis with an elevated gray whitish lesion just under the center of the cornea in his right eye. The lesion had an irregular surface. METHOD: We excised the corneal lesion, and studied the excised corneal lesion morphologically. RESULT: The deposit observed just under the corneal epithelial layer was positive for Congo red staining, and showed dichroism under polarizing microscopy. The deposit also showed a immunoreactivity against anti-human lactoferrin antibody. CONCLUSION: The morphological study proved that the deposits under the corneal lesion were derived from lactoferrin. Long term injury of the corneal surface by trichiasis may lead to the deposition and structural changes of lactoferrin originating from tears. PMID- 12647337 TI - [International clinical diabetic retinopathy and diabetic macular edema disease severity scales]. PMID- 12647338 TI - [The research project: motivation and general methodology]. PMID- 12647339 TI - [Research tools]. PMID- 12647340 TI - [Explained bibliography of the HoNOS Scale]. PMID- 12647341 TI - [Services and recruited patients]. PMID- 12647342 TI - [Diagnosis and severity]. PMID- 12647343 TI - [Patients' cost]. PMID- 12647345 TI - [Quality: necessary, important, persuasive]. PMID- 12647346 TI - [Pathology in Vienna]. PMID- 12647344 TI - [Explanatory model of the cost variability of treating patients: methodology and results]. PMID- 12647347 TI - [Dr. Axel Greiner receives the Rudolph Virchow Prize 2002 from the German Society of Pathology]. PMID- 12647348 TI - [Dynamic and flexible classification of transplant rejection]. AB - Different classification systems of renal allograft rejection are in use. Although, significant progress has been made with the introduction of the widely recognized Banff classification scheme, major problems remain unsolved. Minimal diagnostic threshold levels for tubulo-interstitial rejection are lacking. "Hyperacute, acute and chronic rejection or allograft nephropathy" are ill defined entities and thereby hamper rather than promote the communication between clinicians and pathologists and prevent a better understanding of pathophysiological events. Here, we propose a classification system of renal allograft rejection which is primarily based on well characterized morphological changes. All anatomical compartments of the renal cortex i.e., the tubulo interstitial compartment, arteries and glomeruli, can be affected by rejection individually or in combination. In the subgroups only morphological changes known to be induced by rejection are considered. Such a classification system has, in our opinion, several major advantages. Every group and subgroup can easily be amended by "qualifiers" providing additional immunohistochemical and molecular biological information as they are known today or become available in the future. As knowledge expands, qualifiers may be added or removed without fundamentally changing the entire classification scheme--making the system flexible and dynamic. PMID- 12647349 TI - [Molecular genetic classification of renal cell carcinoma]. PMID- 12647350 TI - [Renal tumors in adults: rare tumors and new tumor entities]. AB - International agreement was reached on the histologic classification of renal epithelial neoplasms during the last years. This classification defines malignant neoplasms as clear-cell (conventional) renal carcinoma, papillary renal carcinoma, chromophobe renal carcinoma, collecting duct carcinoma and renal cell carcinoma, unclassified. Benign neoplasms are papillary adenoma, renal oncocytoma and metanephric nephroadenoma/adenofibroma. Over the past few years, new or rare distinctive kidney tumors have been described. The aim of this review is to present examples of recently recognized clinicopathologic tumor entites and to discuss the value of immunohistochemical and molecular tests for the differential diagnosis. The following tumors will be described: mixed epithelial/stromal renal tumors, primary renal synovial sarcomas, primary renal primitive neuroectodermal tumors, low grade myxoid renal epithelial neoplasms with distal nephron differentiation and epitheloid angiomyolipoma. Detection of SYT-SSX gene fusion transcripts resulting from the t(X;18) and the EWS-FLI-1 gene fusion are described as molecular tests for the diagnosis of renal synovial sarcomas and renal primitive neuroectodermal tumors. Immunohistochemical expression of hormone receptor is helpful to diagnose mixed epithelial/stromal renal tumors. It is important to distinguish these tumor entities from adult Wilms' tumors and sarcomatoid renal cell carcinomas because of a different biological behaviour and different therapeutical approaches. PMID- 12647351 TI - [Molecular genetics of urinary bladder cancer progression]. AB - Extensive cytogenetic and molecular analyses have recently helped to gain a much better understanding of the biology of urinary bladder cancer. Early studies had suggested two different pathways of bladder cancer development, one characterized by chromosome 9 losses and the other by p53 mutations. Subsequent studies have greatly expanded these data. Overall, there is compelling evidence for two entirely different bladder tumor entities. One entity consists of genetically unstable tumors that have many cytogenetic alterations, including p53 alterations in about 50% of the cases. Other cytogenetic alterations that can be found frequently in these tumors are deletions of 8p, 9p, 11p, 11q, and Y as well as gains of 1q, 8q, 17q, and 20q. More than 20 chromosomal regions can undergo high level DNA amplification including 17q21 (HER-2/neu). Phenotypically, these tumors are characterized by a high degree of cytological atypia. Their growth pattern can be non-invasive flat (carcinoma in situ), non-invasive papillary (pTaG3) or invasive (papillary or solid). The other, "benign" bladder cancer entity is composed of tumors with a low level of genetic instability, a low number of cytogenetic alterations, and absence of p53 mutations. Morphologically, these tumors are papillary non-invasive neoplasms with a low degree of cytological atypia. Progression to invasively growing carcinoma is extremely rare. CONCLUSION: Molecular studies have revealed profound genetic differences between invasive and non-invasive bladder cancers. This argues against the previously used combination of pTa and pT1 tumor stages as "superficial bladder cancer". The high frequency of genetic alterations in invasive carcinomas enables a markedly improved detection of bladder cancer cells in voided urine using fluorescence in situ hybridization (FISH) assays. PMID- 12647352 TI - [New WHO classification of urothelial carcinoma of the urinary bladder]. AB - The WHO classification of urothelial carcinomas of the urinary bladder (1999) presents the papillary urothelial neoplasia of low malignant potential (PUNLMP) as a new entity in between the papillomas and the papillary urothelial carcinomas. This neoplasia shows a typical basal palisading, a low mitotic rate, and a low MIB-1-proliferation index. The PUNLMP is said to have an increased risk of development of recurrent papillary lesions with the possibility of malignant transformation. At present, there is an intensive discussion on this new entity. The participants of a meeting on the consensus classification on urothelial tumors held in Ancona in 2000 have meanwhile split in two discussion groups. One favors the new WHO classification with the papillary urothelial carcinomas G I, G II, and G III, but without PUNLMP, whereas the other group favors the consensus classification of 1998 with papillomas, papillary urothelial neoplasia of low malignant potential, and non invasive as well as invasive low-grade and high grade papillary urothelial carcinomas. Future long term prospective studies will show the significance of PUNLMP compared to well differentiated non invasive papillary urothelial urinary bladder carcinoma G I (G Ia). Otherwise, there is no significant difference in the classification of carcinomas and non epithelial lesions compared with the previous classification of 1973. The new WHO does however discriminate the minimally invasive papillary urothelial carcinomas in those with infiltration of the lamina propria above the muscularis mucosae (pT1a), the infiltration of the lamina muscularis mucosae (pT1b), and the extension beyond the muscularis mucosae (pT1c). The recurrence rate increases from stage pT1b. This substaging may be of therapeutical relevance. PMID- 12647353 TI - [WHO classification of testicular tumors]. AB - Twenty years after the first edition (1977), the WHO has presented the updated version of the "Histological typing of testis tumours". The revised classification was necessary because some clinically important new entities have been described in recent years. The most important is obviously the precursor lesion of germ cell tumors, which has been called "intratubular malignant germ cells". Such atypical cells appear in the tubules adjacent to the germ cell tumors, in some few cases (6%) also in the contra lateral healthy gonad and rarely in infertile men (1%). The precursor lesion can progress to franc germ cell tumor starting probably with seminoma, which still maintain the capability of differentiation (pluripotente cells) in all other types of non-seminomatous germ cell tumors. This lesion is missed in germ cell tumors of childhood and in spermatocytic seminomas, both seem to have a histogenetic history rather different from the other germ cell in adults. The categories used in the first classification have been preserved and some new diagnoses added. Most of the new diagnoses are subtypes--called "variants"--of well known tumors. Seminoma with syncytiotrophoblastic cells is a variant which should not be confused with choriocarcinoma. Spermatocytic seminomas are perfectly benign tumors but they become a life threatening disease when combined with sarcomas (new entity). In the group of mature teratomas the "dermoid cyst" appears as a benign subtype mostly observed in children. Unfortunately, however, the old term "teratoma with malignant transformation" was changed to "teratoma with malignant areas" in the 1998 classification. This is a harmless name for an extremely dangerous tumor in which one tissue overgrows the other and gives rise to somatic type sarcomas or carcinomas. Such tumors do not respond like germ cell tumors to the usual chemotherapy. Treatment should be tailored according to that used in standard management of the respective sarcoma or carcinoma. In the comments it is mentioned that the testis carcinoid could be a part of teratoma, but the diagnosis is listed in the group of "miscellaneous" tumors together with tumors of ovarian epithelial type. This is a very questionable decision because the normal testis does not contain neuroendocrine cells from which carcinoids would have to be able to develop. Teratomas, however, show plenty of them producing all kinds of typical hormones. "Large cell calcifying Sertoli cell tumour" has been recently described and can be sporadic or inherited. This morphologically peculiar tumor can be part of the Swiss syndrome also called Carney's complex. The patients have cardiac myxomas, spotty skin pigmentation, hormone active nodular hyperplasia of the adrenals and soft tissue myxomas. The newly appearing "mixed germ cell--sex cord/gonadal stromal tumours, unclassified" has a histology similar to the well known gonadoblastomas. In contrast to gonadoblastoma, however, these tumors occur in testes of genotypic and phenotypic normal males. From the practical, diagnostic point of view the new classification does not contain dramatic changes. For the therapy of germ cell tumor an assessment of risk factors found by the pathologists is extremely important. The most important independent predictors of relapse are tumor invasion of blood or lymph-vessels, absence of yolk sac elements and the presence of an embryonal carcinoma component. In the absence of such predictors a surveillance policy allows some patients to forgo chemotherapy. PMID- 12647354 TI - [Innate and acquired immunity in angiotensin-induced malignant hypertension]. AB - Participation of the immune system in angiotensin (Ang) II-induced endorgan damage is not a conventionally accepted idea. Nevertheless, we have evidence from a double-transgenic rat model of malignant hypertension that Ang II leads not only to activated innate immunity via NF-kB, but also to activated acquired immunity via dendritic cells. By means of surface markers, we observed dendritic cell maturation, migration, and contact with CD4 and CD8 lymphocytes in hearts and kidneys of double-transgenic rats. Treatment with dexamethasone or etanercept provides protection in this model, independent of arterial blood pressure-related effects. Our preliminary data implicate innate immunity and acquired immunity. Both cell-mediated and antibody-mediated effects are involved in the latter in this model. PMID- 12647356 TI - Diabetic microangiopathy: pathology and current understanding of its pathogenesis. AB - Recent drastic increase in diabetic population poses serious problems in both health sciences and socioeconomic conditions. The most important issue in the clinical practice of diabetic patients is the treatment and care of chronic complications. It is not fully clear, however, as to the pathophysiology of diabetic microangiopathy and its pathogenesis. Recent studies on microvessel pathology in diabetic patients and molecular analyses on the diabetic animal models disclosed novel features of the dynamic changes of specific organ pathology affected by diabetes and factors involved in its pathogenesis. Under long-term hyperglycemia, early stimuli elicit adaptive reactions of tissues showing acute inflammatory processes of vessel walls and then late irreversible and regressive changes of microangiopathy. Consequently, remodeling of vascular cells and excessive matrix production are cardinal feature. The precise mechanisms of how these tissue changes occur remain speculative; increased polyol pathway, excessive non-enzymatic glycation, increased protein kinase C activity, as well as oxidative stress are all interrelated for the cause and development of the microangiopathy. PMID- 12647355 TI - [Pathology of vasculitis: differential diagnosis and selected disorders]. AB - The morphological diagnosis of vasculitis is a challenge for the pathologist for many reasons. Primary systemic vasculitides are rare diseases (ann. incidence 4 5/100,000 according to the Vaskulitis-Register Schleswig-Holstein), pathognomonic histopathological findings are hard to obtain in many patients even with multiple biopsies, and the nomenclature of vasculitis has long been confusing and unequivocal. The Chapel-Hill Consensus Conference (1992) is credited with the establishment of clinico-pathological definitions for primary systemic vasculitides. For the discrimination of primary systemic vasculitis from secondary vasculitis as well as pseudovasculitic syndromes, correlation with clinical data is essential; it has been facilitated by the American College of Rheumatology (ACR) classification criteria (1990). Differential diagnostic schemes for vasculitis are based on the predominant type and size of the blood vessel involved, as well as the histopathological type of inflammation and the presence or absence of immune complex deposits. However, differential diagnostic problems remain: even with the clarified nomenclature of Chapel-Hill, considerable overlap still occurs. A clinical diagnosis of vasculitis is now often established in an earlier disease stadium, in part of the cases facilitated by ANCA testing; the pathologist may therefore at first be confronted with a more subtle or abortive morphology, while in the course of disease, histopathology is modulated by immunosuppressive therapy. A number of immunopathogenic mechanisms prevailing in vasculitis can be verified with immunohistochemical and molecular biological methods, although it is frequently not known how they are initiated and perpetuated. ANCA-associated vasculitides serve as a good example to demonstrate the central role of morphology, its potentials but also its limitations in the differential diagnosis of vasculitis. PMID- 12647357 TI - Bronchiolitis obliterans. Organizing pneumonia. AB - Bronchiolitis obliterans combined organizing pneumonia (BOOP), now called organizing pneumonia, is a multi-etiologic disease. It can present as a solitary lesion, or as multinodular or diffuse interstitial lung disease. It is speculated if solitary BOOP may evolve into inflammatory pseudotumor of the lung. BOOP can be seen after non-resolving infectious bronchopneumonia as well as acute interstitial pneumonia with diffuse alveolar damage. BOOP can be the early morphologic pattern in toxic inhalation, especially water-soluble substances, but also in drug induced lung disease. BOOP can be the late stage of extrinsic allergic alveolitis, but also a morphologic sequel of collagen vascular disease. Even Wegener's granulomatosis can be preceded by a BOOP pattern. In many cases a careful analysis of BOOP, including changes of the pneumocytes, macrophages, myofibroblasts and endothelial cells, can establish the correct etiologic diagnosis. For example virus-induced pneumocyte proliferation can be seen months after the onset of interstitial pneumonia, and can be found within BOOP. A small percentage of BOOP, however, has to be labeled as idiopathic, which is important too, because of different modalities of therapy. Idiopathic BOOP also is different with respect to prognosis. In the overview different BOOP etiologies will be discussed, and the etiologic background will be analyzed. The pathogenesis will be discussed with respect to the understanding of the causing mechanisms. The role of bronchoalveolar lavage and the optimal tissue sample for establishing the diagnosis will be discussed and demonstrated by examples. A part of the presentation will deal with the differential diagnosis, such as usual interstitial pneumonia, non-specific interstitial pneumonia, constrictive, and respiratory bronchiolitis combined interstitial lung disease. PMID- 12647358 TI - [Differential diagnosis of amyloidosis]. AB - Amyloid is defined as a proteinaceous tissue deposit that shows a typical green birefringence in polarized light after staining with Congo red, the presence of non-branching linear fibrils of indefinite length with an approximate diameter of 10-12 nm, and a distinct X-ray diffraction pattern consistent with Pauling's model of a cross-beta fibril. Amyloid may deposit locally, organ-limited, tumor like, or may present as a generalized disease. The origin of amyloid is diverse: twenty different fibril proteins have been described so far. The precursor proteins differ from each other in their primary structures and functions. The only common denominator is the propensity to form anti-parallel cross beta fibrils under certain circumstances. Early diagnosis of amyloid is still a major challenge in surgical pathology. Histological proof can be obtained using Congo red staining and polarisation microscopy. However, small deposits may be difficult to discern and sensitivity can be improved by using fluorescence microscopy. Classification of amyloid is mandatory, since amyloid is treatable and different treatment regimens are applied to different amyloid diseases. AA amyloidosis requires intensified treatment of the underlying disease. Patients with generalized AL amyloidosis receive high dose chemotherapy and stem cell transplantation. Familial amyloidoses (AApoAI, AFib, ATTR) may undergo liver transplantation. PMID- 12647359 TI - [WHO classification of breast tumors and tumors of the female genital organs: pathology and genetics]. AB - Under the supervision of P. Kleihues and L. H. Sobin, the "World Health Organisation Classification of Tumours" with its numerous volumes is currently being re-edited. Following the first three volumes on "Central Nervous System", "Digestive System" and "Haematopoetic System", the volume on "Tumours of the Breast and Female Genital Organs" will be published this autumn. This volume will be edited by F. Tavassoli and P. Devilee. The new WHO books serve a double function of text books and atlases of tumour pathology. The first part of the new volume includes the genetics and pathology of the entire spectrum of malignant tumours and their precursor lesions. The following part is devoted to benign proliferative lesions. PMID- 12647360 TI - [Drug-induced liver injury]. AB - Drugs may cause acute or chronic liver damage depending on their mode of action. Hepatotoxic drugs include anaesthetics, psychotropic and anticonvulsant drugs, antiinflammatory agents, steroids, antimicrobial agents and cardiovascular drugs as well as antineoplastic agents. Hepatotoxic agents, including drugs, fall into two categories: (i) intrinsic and obligatory liver toxins with dose-dependent and predictable adverse effects, and (ii) facultative (idiosyncratic) hepatotoxins with non-predictable and non-dose-dependent liver toxicity affecting only few exposed individuals. Intrinsic hepatotoxins may either injure hepatocytes directly, e.g. by direct physicochemical effects, or indirectly by interfering with specific metabolic processes. In the idiosyncratic type of liver injury immunologic hypersensitivity reactions or toxic metabolites may be involved. Clinical and morphologic consequences of adverse drug reactions are acute or chronic liver diseases, including parenchymal damage (finally leading to necrosis or apoptosis), steatosis, cholestasis, various types of vascular alterations, granuloma formation and also neoplastic transformation. Thus, drugs are important causes of liver diseases and may account for up to 40% of cases of hepatitis and up to 25% of fulminant hepatic failure. Moreover, drug-induced injury also plays a leading role as cause of acute cholestasis. PMID- 12647361 TI - [Histopathologic diagnosis of Barrett mucosa and associated neoplasias: results of a consensus conference of the Working Group for Gastroenterological Pathology within the Germany Society of Pathology, September 2001 in Erlangen]. AB - There are a number of difficulties regarding the diagnosis of Barrett's mucosa and the varying grades of neoplasia that may be associated with it. It was therefore the aim of a Consensus-Conference of the Working Group for Gastroenterological Pathology within the German Society of Pathology to achieve standardization regarding the following issues: definition and diagnostic criteria for Barrett's mucosa and its discrimination from intestinal metaplasia of the cardia, diagnostic criteria for intraepithelial neoplasia, number of biopsies necessary to establish the diagnosis, significance of additional immunohistochemical and/or molecular methods as well as importance of a second opinion in the diagnosis of intraepithelial neoplasia. PMID- 12647362 TI - [Brain--music--intelligence]. PMID- 12647363 TI - [Pathogenesis of cystic kidney diseases]. AB - Several gene mutations cause renal cysts in humans, including PKD1 and PKD2 (autosomal dominant polycystic kidney disease, ADPKD), PKHD1 (autosomal recessive polycystic kidney disease, ARPKD), TSC1 and TSC2 (Tuberous Sclerosis), NPHP1 (Nephronophthisis type I), and VHL (von-Hippel-Lindau syndrome). This raises the question, whether there are common denominators of cyst development. Both polycystin-1 and fibrocystin, the gene products of PKD1 and PKHD1, appear to act as adhesion molecules. Polycystin-2, encoded by PKD2, is a ion channel that requires polycystin-1 to translocate to the plasma membrane. Polycystin-1 mandates TSC2 for the transport to the plasma membrane; thus, mutations of TSC2 could cause cyst formation by compromising the function of polycystin-1. Disturbances of fibrillary adhesion complexes may represent the final common pathway of NPHP1 as well as VHL mutations. Delineating common pathways of cystogenesis may help to design therapeutic strategies that combat the development and/or progression of renal cysts. PMID- 12647364 TI - [Pathogenesis of MALT-type lymphoma]. AB - Gastric marginal zone lymphoma, MALT-type (MZBCL) is thought to be derived from precursor lesions in follicular H. pylori-associated gastritis, that through accumulating genetic changes ultimately develop into an autonomously proliferating, monoclonal B-cell lymphoma. In the early phases of the disease, the proliferation is still at least partly dependent on the presence of H. pylori induced T-cell help. Ongoing genetic alterations drive the process into an antigen independent phase. First of all, Fas mutations resulting in loss of function were found in a high frequency in MZBCL and gastric DLBCL and imply that Fas may act as a tumor suppressor gene. Therefore it is possible, that various Fas mutations may arise in mature B-cells during V(D)J recombination and other diversification processes in the course of immune responses as it was shown recently for the bcl-6 gene. Second, allelic imbalances suggest two different pathways of MZBCL development and progression. One group of tumors develops along the pathway determined by the dysregulation of the API2 and MALT1 genes brought about by the t(11;18). These tumors do not accumulate enough secondary genetic aberrations to transform into DLBCL and remain in the stage of MZBCL. Other MALT lymphomas characterized by the absence of the t(11;18) and increased accumulation of various clonal genetic aberrations, most frequently the 3q26.2-27 amplification, could be the source of tumors which eventually do transform into high-grade DLBCL. PMID- 12647365 TI - Transcription profiling of renal cell carcinoma. AB - AIMS: Our aim was to prepare a comprehensive catalogue of the changes in gene expression accompanying the development and progression of renal cell carcinoma, and to correlate these with histo-pathological, cytogenetic and clinical findings. METHODS: mRNA samples from paired neoplastic and non-cancerous human kidney tissue were labeled and hybridized in duplicate against high-density cDNA arrays. Two array technologies were used: 31,500-element transcriptome-wide nylon arrays for hybridization with 37 radioactively labelled sample pairs, and 4200 element kidney- and cancer-specific glass microarrays for hybridization with 19 fluorescently labelled sample pairs. RESULTS: We identified more than 1700 cDNA clones that show differential transcription levels in kidney tumor tissue compared to normal kidney tissue. The functional classification of 389 annotated genes provided views of the changes in the activities of specific biological processes in renal cancer. Among the biological processes with a large proportion of up-regulated genes we found cell adhesion, signal transduction, and nucleotide metabolism. Down-regulated processes included small molecule transport, ion homeostasis, and oxygen and radical metabolism. Furthermore, we explored the feasibility of molecular diagnosis for renal cell tumors using cDNA microarrays on glass slides, investigating the association of transcription levels with tumor type, progression, and a putative prognostic variable. The experimental data is available from the GEO gene expression database (http://www.ncbi.nlm.nih.gov/geo; accession no. GSE3), and a comprehensive presentation of the results is available in the web supplement (http://www.dkfz-heidelberg.de/abt0840/whuber/rcc). CONCLUSION: Transcription profiling using high-density cDNA arrays is a powerful method with the potential to improve cancer diagnosis and prognosis. The identification and classification of differentially transcribed genes, as described in our study, is the beginning of a more complete understanding of kidney cancer. PMID- 12647366 TI - [Gene expression profiling in prostatic cancer]. AB - Basic aspects of the biology and molecular alterations in prostate carcinoma remain poorly understood. New diagnostic and prognostic markers for prostate carcinoma may add additional information to current histopathological diagnosis. In order to achieve these goals, a comprehensive gene expression analysis was performed on non-metastasizing, untreated prostate cancer tissues. RNA expression profiles of approximately 12,600 sequences from 26 human prostate tissues (17 adenocarcinomas and 9 normal adjacent to cancer tissues) were investigated using high-density oligonucleotide microarray technology (Affymetrix). We identified 63 genes which were significantly increased (at least 2.5-fold) and 153 genes which were decreased (at least 2.5-fold). Upregulated genes included several which had not yet been described, such as the genes encoding the specific granule protein (SGP28), several members of the histone family, and the alpha-methylacyl-CoA racemase, but also previously reported ones such as hepsin, LIM domain kinase 2, and carcinoma-associated antigen GA733-2. Laser capture-microdissection of epithelial and stromal compartments from cancer and histologically normal specimens followed by an amplification protocol for low amounts of RNA (< 0.1 microgram) allowed us to distinguish between gene expression profiles characteristic of epithelial cells and those typical of stroma. Most of the genes identified in bulk tumor material as upregulated were indeed overexpressed in cancerous epithelium rather than in the stromal compartment. DNA microarray data for up- and downregulated genes were confirmed by quantitative RT-PCR. We demonstrated that development of prostate cancer is associated with downregulation as well as upregulation of genes that show complex differential regulation in epithelia and stroma. Some of the alterations in gene expression identified in this study may prove useful in development of novel diagnostic and therapeutic strategies. Gene expression profiling of microdissected tumor cells in prostate biopsies may supplement histopathologic diagnosis. PMID- 12647367 TI - [HER-2 and TOP 2A gene amplifications in urinary bladder carcinoma]. AB - The HER-2/neu gene is frequently amplified in bladder cancer. Topoisomerase 2 Alpha (TOP2A) which is located nearby the HER-2/neu gene is an important molecular target for several anti cancer drugs. The frequency of TOP2A amplification in urinary bladder cancer is unknown. It was the aim of this study to determine the frequency of HER-2 and TOP2A amplification in urinary bladder cancer and to evaluate the association of these amplifications with tumor phenotype. For this purpose a tissue microarray containing 768 pTa, 425 pT1 and 571 pT2-4 carcinomas was analyzed by fluorescence in situ hybridization (FISH). Amplifications of both genes were significantly associated with advanced tumor stage and high grade. HER-2 amplification was found in 1.6% of pTa, 7.2% of pT1 and 13.8% of pT2-4 carcinomas (p < 0.0001). HER-2 amplification was present in only 1.1% of grade 1 and 0.8% of grade 2 tumors but in 14.2% of grade 3 tumors (p < 0.0001). TOP2A amplification was present in 0.7% pTa, 1.8% pT1 and 3.4% pT2-4 carcinomas (p < 0.0001). TOP2A was found in none of the grade 1, in 0.2% of grade 2 and 3.8% of grade 3 tumors (p < 0.0001). 1% of all analyzed tumors had simultaneously high level amplification of TOP2A and HER-2. Amplification of both genes were significantly associated with tumor specific survival if all tumors were analyzed together. Given the high frequency of HER-2 amplification in urinary bladder cancer, some of these tumors may respond favorable to Herceptin therapy. The TOP2A amplification status may influence response to anthracyclin treatment. PMID- 12647368 TI - [List of members]. PMID- 12647369 TI - Eight questions consumers should ask on the threat of mad cow disease. PMID- 12647370 TI - Influenza. PMID- 12647371 TI - Smallpox--no longer is it only a disease of historical interest. PMID- 12647372 TI - Death at the Hiawatha Asylum for Insane Indians. AB - From 1903 to 1933, the Hiawatha Asylum for Insane Indians housed hundreds of inmates in Canton, SD. It closed in 1933 in a cloud of scandal and controversy. Death certificate data from deceased inmates were analyzed for diagnostic cause of death and for age. These findings were compared to a sample of non-Asylum adult deaths for the same years in the same county. The inmates died younger (average age-42), and of tuberculosis (63 of 182 deaths). The historical import of this is discussed. PMID- 12647373 TI - "Doctor, oh, by the way ..." a 58-year old woman with muscle aches. PMID- 12647374 TI - State of South Dakota's child: 2002. AB - In 2001, for the second consecutive year, there was an increase in births in South Dakota that also included the highest rate of low birth weight infants born in 28 years. The infant mortality rate also increased from its exceptionally low rate in 2000. The rate of Sudden Infant Death Syndrome in South Dakota is over twice that noted nationally and emphasizes the need for vigilant efforts to promote substance free pregnancies and safe sleeping for infants. The special topic of this year's report is emergent literacy. This concept reveals the continuum of experiences that begin in infancy and contribute to a child's learning how to read and write. Efforts that have successfully promoted reading with well-baby and child health care visits are described. PMID- 12647376 TI - [Usefulness of PET scan guidance in stereotaxic radioneurosurgery using a gamma knife]. AB - Integration of the metabolic data of PET in the dosimetry planning of radiosurgical treatment with the Gamma Knife, can optimize the definition of the target volume for infiltrating or ill-defined brain tumors. Our experience of twenty patients treated with this technique demonstrates the interest of this original approach that should improve the results of radiosurgical treatment of cerebral tumors. PMID- 12647375 TI - [Metabolic interactions between glucose and fructose]. AB - Fructose-1-phosphate, the product of fructose phosphorylation by fructokinase, favours glucose phosphorylation by glucokinase, at the intervention of a regulatory protein. Glucose confers to glucokinase positive cooperativity, towards fructose. These reciprocal effects are operative in hepatocytes and pancreatic islets. They could optimize their glucostatic function. PMID- 12647377 TI - [Oxytocin: a natural means of treating psychological stress]. AB - Oxytocin as an endogenous antistress hormone. The neurohypophysial peptide oxytocin (OT), besides it well known uterotonic and milk ejection activity, share also an inhibitory action on corticotrope activity. This has been demonstrated not only in pharmacological (perfusion) but also in physiological (lactation) conditions. This action is opposite to that of its "sister" hormone vasopressin (AVP) thus bringing arguments favouring the ago-antagonist ying-yang hypothesis. A non pharmacological stimulation of ocytocine secretion, as we have recently demonstrated in a preliminary study, related to hypnosis, could induce a beneficial inhibition of corticotrop axis and needs further rigorous experimental approach. PMID- 12647378 TI - [Multiple sclerosis: from the immune system to inflammatory demyelination and irreversible neurodegeneration]. AB - The complex pathogenesis of multiple sclerosis includes inflammation and potentially disabling focal lesions that are associated with heterogeneous, often destructive pathologic changes disseminated throughout the white matter of the central nervous system. Genetic and environmental factors may trigger the movement of autoreactive T cells, from the systemic circulation into the central nervous system through disruption of the blood-brain barrier. In the brain, local factors may enhance the immune reaction, resulting in the proliferation of pro inflammatory CD4+ type 1 T cells, and ultimately in immune-mediated injury of oligodendrocytes and myelin. The exposed axon segments may be susceptible to further injury from soluble mediators such as cytokines, chemokines, complement and proteases, resulting in irreversible axonal lesions. Partial remyclination may occur by differentiation of resident oligodendrocyte precursor cells. Decreases in the volume of the brain and spinal cord, indicating atrophy, correlate with both the loss of axons and the occurrence of extensive demyelination, and are markers of the degenerative phase of the disease. Besides classical motor and sensory signs and symptoms, cognitive impairment is frequently observed and increases the burden of the disease. PMID- 12647380 TI - Fundamentals of protein structure. PMID- 12647379 TI - Defining bioinformatics and structural bioinformatics. PMID- 12647381 TI - Fundamentals of DNA and RNA structure. PMID- 12647383 TI - Macromolecular structure determination by NMR spectroscopy. PMID- 12647382 TI - Computational aspects of high-throughput crystallographic macromolecular structure determination. PMID- 12647385 TI - Molecular visualization. PMID- 12647384 TI - Electron microscopy. PMID- 12647386 TI - The PDB format, mmCIF, and other data formats. PMID- 12647388 TI - Other structure-based databases. PMID- 12647387 TI - The Protein Data Bank. PMID- 12647389 TI - Protein structure evolution and the SCOP database. PMID- 12647390 TI - The CATH domain structure database. PMID- 12647392 TI - All-atom contacts: a new approach to structure validation. PMID- 12647391 TI - Structural quality assurance. PMID- 12647393 TI - Structure comparison and alignment. PMID- 12647394 TI - Secondary structure assignment. PMID- 12647395 TI - Identifying structural domains in proteins. PMID- 12647396 TI - Inferring protein function from structure. PMID- 12647397 TI - Prediction of protein-protein interactions from evolutionary information. PMID- 12647398 TI - Electrostatic interactions. PMID- 12647399 TI - Principles and methods of docking and ligand design. PMID- 12647400 TI - Structural bioinformatics in drug discovery. PMID- 12647401 TI - CASP and CAFASP experiments and their findings. AB - This short introductory chapter is intended simply to introduce a sense of the progress, limitations, challenges, and likely future developments in the field of protein structure prediction through what seems to be a unique scientific process. CASP and CAFASP represent a direct challenge and careful assessment of a field of study that has captured the interest of many scientists. Three of the best scientists in the field and their colleagues provide a more detailed description of the field and how it is developing in Chapters 25, 26, and 27. As prediction methods have advanced the distinction between comparative modeling, fold recognition, and novel fold recognition have blurred somewhat. It is a testament to the community that as the knowledge of the algorithms evolved, World Wide Web servers providing access to these algorithms appeared. Thus, making it relatively straightforward for any investigator to apply a melting pot of methods to the prediction process. What all approaches need are more targets and a continued refinement to the evaluation process. The first need is being met in part by the PDB, which is, with depositors' approval, releasing sequences ahead of structure release (see http://www.rcsb.org/pdb/status.html). Further, the structural genomics projects are reporting their progress for all targets on a weekly basis (see http://targetdb.pdb.org/). While there is no indication that the sequences of the latter will lead to a structure, it is a rich source of targets (17,000 in October 2002). Not only do CASP and CAFASP measure progress, they help define where efforts should be directed to move the field forward. It is a testament to how far the field has come that investigators are now turning to the unknown. Although attempting to predict a structure that will appear experimentally helps improve the methods applied to structure prediction, it does not further our understanding of living systems directly. Attempts at defining the "The Most Wanted" (Abbott, 2001)--the structures most in need of prediction to help further our understanding of the biology, and the efforts to make those predictions, speak to a healthy future for the field of protein structure prediction. To the many individuals who help define the CASP and CAFASP processes, serve the community as assesors and compete in the experiments this is a tribute. PMID- 12647402 TI - Homology modeling. PMID- 12647403 TI - Fold recognition methods. AB - We are still missing a basic understanding of sequence/structure/function relationships in proteins. Analogy-based prediction algorithms remain the only reliable fold prediction tools. New methods, such as threading and hybrid threading/sequence fold recognition, can often recognize even the most distant homologues and, in some cases, even unrelated proteins with similar overall structures. This knowledge pushed the envelope of analogy-based function analysis to the point that the majority of newly sequenced genomes can be tentatively assigned to already characterized protein superfamilies. However, at this evolutionary distance, fold prediction is no longer equivalent to function prediction. Instead of having the same exact function, distantly related proteins might share some functional analogy that is not obvious to the casual observer. The main challenge facing the fold recognition field is to develop tools to follow the structure prediction with function prediction and analysis. PMID- 12647404 TI - Ab initio methods. PMID- 12647405 TI - Prediction in 1D: secondary structure, membrane helices, and accessibility. PMID- 12647406 TI - Structural genomics. PMID- 12647407 TI - Diagnostic approach to Vater's papilla tumours. PMID- 12647408 TI - How accurate is preoperative diagnosis by endoscopic biopsies in ampullary tumours? AB - INTRODUCTION: The benign or malignant nature of ampullary tumours has prognostic and therapeutic implications. On the other hand the difficulty of reaching a correct preoperative diagnosis in these lesions is well known, even when we have a histological study obtained by endoscopic biopsies. MATERIAL AND METHODS: We review all ERCP's in which biopsies of the papilla were taken, performed between January 1991 and September 2000. We analyse the concordance rate between preoperative diagnosis (endoscopic biopsies obtained during ERCP) and definitive diagnosis (surgical specimen) (n = 32), and the possible influence of previous sphincterotomy in our results. RESULTS: In the 32 patients studied global accuracy between pre- and postsurgical diagnosis was 68.7%. This accuracy was markedly higher in the group with ampullary cancer (82.7%) than in the group with ampullary adenoma (50%) (p = 0.12). In the group of patients with sphincterotomy accuracy was 56.25% and increased up to 81.25% in the group of patients without sphincterotomy, although statistical significance was not reached (p = 0.25). CONCLUSIONS: In our series, the accuracy of endoscopic biopsies is higher in the adenocarcinoma group than in the adenoma group, obtaining better results in patients without previous sphincterotomy. The impossibility of a preoperative and absolutely certain confirmation of the benign or malignant nature of ampullary tumours forces us to be cautious in deciding type of surgical resection. PMID- 12647409 TI - Helical CT evaluation in the preoperative staging of gastric adenocarcinoma. AB - INTRODUCTION: The aim of this study is to evaluate helical CT as a basic preoperative test for the staging of gastric adenocarcinoma. MATERIAL AND METHOD: We enrolled 50 patients with a diagnosis of gastric adenocarcinoma, evaluated helical CT scans, and compared their findings to the microscopic, pathology confirmed ones obtained by laparotomy. UICC's TNM classification, 5th edition (May 1997), was used. RESULTS: Best results regarding Sensitivity were obtained in the diagnosis of lymph node disease (83%; 69-92%); regarding specificity we obtained 92% (79-98%) for T4, and 89% (77-96%) in metastasis detection. Accuracy was 70% for T1-T2, and 62% for T3. CONCLUSIONS: 1. For the T category, helical CT effectively detects advanced cases classified as non-resectable. 2. For the N category, helical CT prediction is better than ever before with previous CT generations. 3. For the M category, helical CT is a useful test to identify the presence of liver metastases that would render surgery unnecessary. PMID- 12647410 TI - Helical CT of the stomach: differentiation between benign and malignant pathologies, together with the staging of gastric carcinoma. AB - OBJECTIVES: To evaluate the capacity of Helical Hydro-CT to differentiate between benign and malignant gastric pathologies, and also to measure its usefulness in the staging of gastric carcinoma. PATIENTS AND DESIGN: We perform a cross sectional study to evaluate the diagnostic efficacy of CT, including patients prospectively. We study 92 patients with a clinical suspicion of gastric pathology using helical CT with a contrast agent (HCTC), water being the oral contrast agent, and i.v. iodine contrast. According to the findings of previous works, we considered stomachs to be normal when the thickness of their wall was less than or equal to 6 mm, with a multilayered appearance that stands out homogeneously with the i.v. contrast. A malignant tumour was diagnosed if the thickness of the wall was greater, together with strong marking by the contrast agent and loss of the normal multilayered pattern. Parietal thickening was classified as gastritis if there was no excessive marking and no loss of the layered pattern. Masses with smooth borders, intraluminal growth and a rounded morphology were diagnosed as sub-mucosal tumours. The results of our 92 studies were compared in all cases with the findings of endoscopic studies, while in the 52 patients treated with surgery they were compared against surgical findings. RESULTS: In 12 of the 92 patients studied using HCTC no gastric pathology was observed by CT or endoscopy. Of the 80 pathological cases CT was used to diagnose 29 as benign pathology, 19 of which were confirmed as such by histology, and 51 cases as malignant pathology, of which 49 were confirmed by histology. We obtained a sensitivity of 81.7% in the diagnosis of malignant pathology and a specificity of 90%, with a PPV of 96% and a NPV of 62%. Regarding TNM staging (in comparison with the 1997 TNM classification), the diagnostic reliability obtained amounted to 56% for T and N, and 87% for M. CONCLUSIONS: Helical hydro CT makes it possible to diagnose advanced gastric carcinoma. Its usefulness basically lies in the evaluation of metastatic neoplastic disease. It has also been shown to be useful in the diagnosis of benign pathologies. It is not a good screening method for the diagnosis of gastric carcinoma. PMID- 12647411 TI - Chronic viral hepatitis: liver biopsy, yes or no? AB - LB increases costs and has--although minimal--potential risks; however, LB is: a) the gold standard to get to know lesion severity (grade and stage); b) the only method for the diagnosis of an F3 stage, which progresses to liver cirrhosis in approximately 10 years; c) the only method for the certain diagnosis of compensated liver cirrhosis, which requires early diagnosis programs for oesophageal varices and hepatocarcinoma; and d) the only method for the certain diagnosis of lesions predicting a good therapeutic response. On all these grounds -save for exceptions (contraindications, clinical and/or ultrasonographic cirrhosis)--liver biopsy should be carried out in the initial study of all patients. In non-responders to anti-viral therapy liver biopsy should be repeated once or twice with 5-10-year intervals to ascertain fibrosis growth rate and, according to this, plan a most appropriate follow-up. Assuming hepatitis progresses rapidly in all HIV co-infected patients, anti-viral therapy could be administered with no previous LB; such biopsy would be performed in non responders to decide their course of therapy. PMID- 12647412 TI - Extra-gastrointestinal stromal tumour--semiology and clinical therapy peculiarities. AB - The gastrointestinal stromal tumor with extragastrointestinal location are very infrequent. We often diagnose them when they show a big size. Their bening or malignant nature is difficult to fix. The best histological parameters to evaluate their prognosis are a high cellularity, the tumor-like necrosis presence and having more than two mitosis per fifty high-power fields. We introduce an asyntomatic patient's case by a routine echographical control for chronic hepatitis by C virus, that has been diagnosed of a mesentery tumor. The patient has been treated surgically. The inmunohistological study of the tumor had confirmed a stromal gastrointestinal tumor. The showed case's analysis and the considered bibliography suggest some clinical discoveries characteristic of this entity. The histogenesis of these neoplasias are examined and made up to date and the usefulness of the new medication to control check the tumor-like progress is emphasized. PMID- 12647413 TI - [Celiac disease]. PMID- 12647414 TI - [Subcapsular hematoma as complication of percutaneous hepatic biopsy]. PMID- 12647415 TI - [Pharmacogenetic study of thiopurine S-methyltransferase (TPMT) and thiopurine toxicity]. PMID- 12647416 TI - [Pancreatic abscess drained through the inguinal canal]. PMID- 12647417 TI - [Hepatic angiomyolipoma]. PMID- 12647418 TI - [Short biography of professor Tadeusz Bilikiewicz]. PMID- 12647419 TI - [Works of T. Bilikiewicz as an example of influential scientific ideas]. PMID- 12647420 TI - [Etioepigenetic nosography by Tadeusz Bilikiewicz as one of the earliest multiaxial psychiatric classification]. PMID- 12647421 TI - [Professor Tadeusz Bilikiewicz as an historian of medicine]. PMID- 12647422 TI - [Anticonvulsants (carbamazepine, valproate, lamotrigine) in bipolar affective disorder]. AB - The introduction of anticonvulsants to the treatment of recurring affective disorders, has significantly broadened the possibilities of helping the ill, especially in the prophylaxis of recurring symptoms of bipolar affective disorders. Carbamazepine and valproate (probably also lamotrigine) are a valuable alternative of Lithium Carbonate. The application of anticonvulsants requires strict rule regiment, which includes: gradual dose increase (especially in lamotrigine), gradual dose reduction when planning ending of the therapy, drug blood-level monitoring, liver action monitoring, haemopoietic system monitoring, in the case of the appearance of flu-like symptoms and skin changes--withdrawal of the drug. In simultaneous application of other drugs (e.g. psychotropic), probable interactions should be considered. PMID- 12647423 TI - [Long-term lithium prophylaxis in the bipolar affective disorder]. AB - The purpose of the study was to evaluate the efficacy of lithium prophylaxis in patients with bipolar affective disorder and to determine clinical differences between lithium responders and non-responders. Clinical and demographic data of 103 patients treated with lithium for at least 10 years were analysed. 24 patients had no episodes on lithium (excellent responders--ER), in 48--at least 50% reduction of episode index (number of episodes/year) was observed, and the remaining 31 did not benefit from lithium treatment. Duration of illness before lithium introduction was significantly longer in lithium non-responders. Lithium responders had significantly higher episode index before lithium. Percentage of excellent responders diminished during treatment. The trend toward higher lithium plasma level in excellent responders was observed. Non-responders had significantly more depressions during first year of lithium prophylaxis. Depressive episodes during first year of treatment may be a predictor of non response. PMID- 12647424 TI - [Therapeutic drug monitoring in depression--new perspectives]. AB - 34 patients with diagnosis of depressive episode (ICD-10) were treated for 8 weeks with tricyclic antidepressants (TCA) 22 patients were treated with clomipramine 75-175 mg daily, 11 with imipramine 75-150 mg and 1 with amitriptyline 150 mg. Following parameters were analysed: plasma concentration (FPIA, HPLC), pharmaco-EEG (spectrum power for delta, theta, alfa1, 2, beta 1, 2, 3 by the use of FFT), clinical improvement (HAMD, HARS, SGI, SERS). 50% reduction in HAMD was regarded as improvement. RESULTS: No relationship between mental state and plasma concentration of TCA was found, initial results in SERS were prognostic for the course of treatment, pharmaco-EEG was typical for antidepressants after two weeks of treatment and reflects clinical improvement and stabilization of plasma concentration. Comparing the plasma concentration of TCA measured by the use of FPIA and HPLC method may be useful for the implementation of monitoring therapy. PMID- 12647425 TI - [The comparative studies of therapeutic results in patients with psychotic refractory depression treated with selected antidepressants and unilateral ECT]. AB - Out of 14 patients with severe, refractory, psychotic depression-7 patients were treated by UECT and the rest with amitriptyline or mianserin. Remission and improvement were recorded in 6 patients after a course of UECT but in none treated with antidepressants. PMID- 12647426 TI - [Intravenous clomipramine for depressive disorders]. AB - 51 patients with depressive episode (40--recurrent depressive disorder, 11- bipolar affective disorder) were treated by the clomipramine intravenous infusions. The number of infusions varied from 5 to 21 (mostly 10-16). The median of maximal dose was 150 mg. There were significant clinical improvement measured by CGI. The therapy was well tolerated. PMID- 12647427 TI - [Prognostic value of pharmaco-EEG in depression treated with moclobemide]. AB - Ten women (age 41.90 +/- 5.67) with depressive episode (moderate, mild; ICD-10) were treated with moclobemide (450 mg). After 4 weeks two groups (each consists of 5 patients) with different response to the drug emerged (responders: improvement in Beck Inventory of Depression 13.6 +/- 2.9 and nonresponders: 2.0 +/- 2.4). FFT analysis of their EEG (taken before and after 4 weeks of treatment) showed some relative power differences in most bands of frequencies between groups, mostly marked for a (7-14 Hz), smaller for a and a, but no differences for e were found. Alfa activity was relatively low after treatment in nonresponders and slightly higher in responders. However any of these results had statistical value. PMID- 12647428 TI - [Phototherapy as a supporting treatment in depressive patients]. AB - Effects of treatment of depression with combined antidepressants and phototherapy and antidepressants alone were compared. Phototherapy was applied 5 days a week, up to 1.5 hour a day, 4500 Lux approximately. Mental status was evaluated by the use of HAMD and BDI in the beginning and after two weeks of treatment. Final results were similar in both groups but in the group treated with combined therapy significant improvement just after the first week of treatment was seen. It can be suggested that phototherapy when applied together with antidepressants may accelerate remission. Particularly good response was seen in patients with bipolar affective disorder. Influence of two kinds of treatment on particular symptoms of depression was different. In patients who were given only antidepressants improvement of sleep and appetite was observed first. Patients treated also with phototherapy reported increase of energy and mood improvement. PMID- 12647429 TI - [Prognostic value of pharmaco-EEG in depression treated with fluvoxamine]. AB - 14 patients (age 42.1 +/- 15.5; 12 women, 2 men) with a depressive episode (moderate, mild; ICD-10) were treated with fluvoxamine (150 mg). After 4 weeks two groups with different responses to the drug emerged (10 responders: improvement in Beck Inventory of Depression 13.6 +/- 2.9 and 4 nonresponders: 2.0 +/- 2.4). FFT analysis of their EEG (taken before and after the treatment) showed some relative power differences in most bands of frequencies between groups, mostly marked for a (7-14 Hz) and smaller for the others. Alpha activity was relatively low after treatment in nonresponders and slightly higher in responders. However, none of these results had statistical value. PMID- 12647430 TI - [Assessment of efficacy and tolerability of mirtazapine treatment of patients with depression]. AB - The paper presents results of the study on the efficacy and tolerability of mirtazapine treatment of patients with depression. The study was open and conducted in three centers. 65 out- and in-patients with diagnosis of depression was included. 50% reduction of the score in the HAM-D scale was obtained in 57%, according to MADRS scale--in 61% patients. Side effects were reported in 49% patients. The most frequent side effects were: drowsiness, fatigue, anxiety, sleepiness, weight increase, headache. The results of the study are comparable with other mirtazapine studies. PMID- 12647431 TI - [Mirtazapine--an antidepressant]. AB - The pharmacological properties and clinical value of mirtazapine has been presented, based on a literature review. It is concluded that mirtazapine has syndromolytic anti-depressive actions towards syndromes of various clinical picture and intensity. It is particularly highly recommended in the treatment of depressive disorders with insomnia, anxiety and body-weight reduction. It is a safe drug, which can be used, in long-term supportive treatment. PMID- 12647433 TI - [Efficacy and tolerance assessment of mitrazapine in the treatment of depression in the elderly--preliminary report]. AB - Depressive disorders are often met in the elderly, its course is usually protracted and outcome sometimes fatal. The purpose of this study is evaluation of efficacy and tolerance of mirtazapine in this group of patients. 24 patients (mean age 73.8) with major depressive episode (> 20 in 17-tem HAMD; also CGI scale was used for evaluation) without psychotic features were treated with mirtazapine (30-45 mg) for 6 months. RESULTS: Improvement (especially such symptoms as better mood and sleep, less anxiety) could be seen in 75% of patients. PMID- 12647432 TI - [Acetylcholinesterase inhibitors--beyond Alzheimer's disease]. AB - Based on a literature review, the application of Acetylcholinesterase inhibitors, IAchE (donepezil, rivastigmine, galantanine) in the treatment of various illnesses which have cholinergic system disability and dementia in their course- (dementia with Lewy bodies, vascular dementia, Parkinson's disease, Multiple Sclerosis, Down Syndrome), delirium symptoms (e.g. Korsakoff psychosis), hyperkinesis, attention and memory disorders--is presented. Promising results in the treatment of late dyskinesias, in schizophrenia with impaired cognitive function, as well as in the additional treatment of various psychotic states are noted. It should be stressed that in Poland, the IAchE have been approved only in the treatment of slight to moderate dementia in the course of Alzheimer's disease. PMID- 12647435 TI - [Electroconvulsive therapy in the treatment of depression in the elderly]. AB - In this paper we discuss seven cases of major depression in elderly (age > 60) treated with ECT. Six suffered from unipolar disorder and one from bipolar; four of them had psychotic features. In all patients at least some improvement was gained. Memory impairment appeared in two patients, however in one case no abnormalities in neuropsychological tests were found; in the second patient memory impairment, which could have been seen before treatment, became slightly more pronounced. In one patient after injection used for anaesthesia convulsion (type grand mal) occurred, but when the medicine was changed all ECT sessions were successful without any side effects. In three patients ECT had been successfully used before and was still efficient during our study. In other three patients ECT as prevention is still being used. Our first experience with ECT in elderly patients is highly encouraging, especially after careful selection of patients. PMID- 12647434 TI - [Efficacy of selected antidepressants in senile depressive disorders--the author's own research]. AB - Apart from dementia, depressive disorders are the commonest psychiatric disorders of elderly people. The treatment of these patients encounters numerous limitations due to their susceptibility to undesirable effects. Therefore, new better drugs are being searched for, which would be as effective as tricyclic (TLPD) antidepressants. The purpose of the research was to assess the effectiveness of selected antidepressants in senile depressive disorders. 90 patients (76 women and 14 men), diagnosed with a depressive episode according to the ICD-10 criteria, were examined. The following scales were used: the MMSE, Hachinski, IADL, GDS, ADAS and GdeprS. The CGI scale was used to assess clinical improvement. The research showed that mianserin was as effective as the TLPD. Both the TLPD and mianserin proved to be more efficient than the SSRI. PMID- 12647436 TI - [The cognitive deficits in the late-life depression and their prognostic value for pharmacotherapy]. AB - The cognitive deficits in the late-life depression are considered as risk factor for presentation of dementia in the long-term prognosis. In this research we were looking for correlations between the cognitive deficits and depression, their influence for the short-term prognosis and the activity of daily living in the elderly. 90 patients with depression (ICD-10 criteria were used) were assessed with scales: MADRS, MMSE, ADAS, IADL. After 3 months the evaluation with clinical improvement scale was made. The results indicate for correlations between the cognitive deficits and intensity of depression. No influence of cognitive deficits level for the clinical improvement after 3 months was proved. The intensity of depression was connected with lower level of daily living activities (assessed with IADL). PMID- 12647437 TI - [Specificity of depression treatment in elderly population]. AB - Aging of population is one of the most important reasons for the increased incidence of depression observed in recent years. In 1994, 11% of Polish population aged 65 years or more. A similar tendency has been observed in other countries. These data prove the importance of depression in elderly people and its significance as a clinical as well as social problem. Depression in the elderly, in comparison to younger people, characterizes with a more variable and, usually, a more complex etiology. It is difficult to estimate the influence of aging, with all accompanying physical consequences, on the development of depression. Changes connected with aging are considered factors contributing to development of this disease and not its direct reason. According to Puzynski (1979), there are two major clinical patterns of depression in the elderly: depression with delusions and motional anxiety and depressive-astenic syndrome. The diagnosis of depression in the elderly is not easy. Classical symptoms of depression are becoming less clear and distinctive with aging. When treating depressive elderly people, it is important to remember about psychosocial influence, social therapy and psychotherapy directed to improve patient's self esteem and create a positive image of future and surrounding world. PMID- 12647438 TI - [Geriatric Depression Scale as auxiliary diagnostic tool used in patients 55 years and older]. AB - The purpose of this study was to examine the Geriatric Depression Scale translated into Polish for its sensitivity and specificity in relation to obligatory criterions ICD-10. 208 at random select patients from Psychiatry Department and Psychiatry Clinic were included into study. Diagnosis was based on ICD 10 criteria. Geriatric Depression Scale were used in full version containing 30 of questions directed to examined (self-rating). Each questions was read to examined persons. At last 185 persons were included to statistical analysis's. Sensitivity and specificity obtained for critical value equal or higher from 11 points, were 81% and 47% Geriatric Depression Scale is useful tool in initial diagnosis of depression in older people. However one should be clearly to underline, that investigation with Geriatric Depression Scale should start diagnostic process, never while to take place full psychiatric investigations. PMID- 12647440 TI - [Study of the protein glycation in Alzheimer's disease]. AB - Advancement in research on glication process in pathogenesis of AD is presented. Its results may open new, promising possibilities of treatment of neurodegenerative diseases including AD with inhibitors of glication. PMID- 12647439 TI - [Estimation of the risk of Alzheimer type dementia based on "Prognostic Questionnaire"]. AB - The aim of undertaken studies was verification of usefulness of "Prognostic Questionnaire" for estimations of risk of Alzheimer type dementia. In support for own base were included 286 persons, which were investigated by scales creating of "Prognostic Questionnaire" in years 1991-1993. From among 286 persons originally classified to renewed study one surrendered 163 persons. Result in investigation MMSE equal or lower from 25 points was base to removal full psychiatric investigations which had in view recognition of Alzheimer type dementia or its exclusion. Possible Alzheimer disease were recognized at 23 persons. Sensitivity and specificity for "Prognostic Questionnaire" obtained for several critical values were marked. The study showed usefulness of joint interpretation of results of several clinical scales creating "Prognostic Questionnaire" for estimations of risk of Alzheimer type dementia. PMID- 12647441 TI - [Second generation of neuroleptics--expectations and the reality]. AB - The expectations regarding the successful treatment of psychotic disorders, especially schizophrenia, are quite high. By introduction of the antipsychotic drugs of second generation, the hope of better efficacy and tolerability has arisen. Up to now, basing on the clinical experience it seems that the efficacy of II generation antipsychotics reflects not only the positive but also the negative as well as affective schizophrenic symptoms. These drugs do not worsen the cognitive functioning of patients, but sometimes even improve some cognitive functions. Their tolerability is in general better in comparison to the classic neuroleptics, but sometimes it happens that unfavorable drug-induced side-effects during treatment (e.g. extrapyramidal symptoms) may occur. The careful selection of the drug and its dosage increases the possibility of good clinical improvement, but more wider use of these drugs is recommended in order to evaluate their practical usefulness. PMID- 12647442 TI - [The treatment of schizophrenia--improper choices, adverse events. Preliminary report]. AB - Files of 68 patients with paranoid schizophrenia treated first time in Department of Psychiatry Medical University of Lublin and in Psychoneurological Hospital of Lublin were analyzed. The aim of this study was evaluation of treatment according to the standards. In the most cases treatment was in consent with standards regarding choice of neuroleptic, period of time of treatment and its effectiveness. The only objections, which we founded were: 1) strong neuroleptics in therapy were used too early and, 2) doses of neuroleptics were elevated to fast, 3) neuroleptics in depo form were used very rarely, 4) atypical neuroleptics were used also very rarely. PMID- 12647444 TI - [Olanzapine in the first episode of schizophrenia in young adults]. AB - The study concerns the therapeutic effectiveness of olanzapine in the treatment of the first episode of schizophrenia in children and young adults from the 12-19 year old age group. The positive symptoms were the first ones to cease. A constant rise in effectiveness was noted with time duration of application, especially in the case of negative symptoms. The treatment with olanzapine enabled most of the youngsters to continue school. PMID- 12647443 TI - [Factors connected with the effect of risperidone on psychopathological symptoms and working memory in schizophrenia]. AB - The aim of the study was to assess the impact of some clinical factors such as family history of psychiatric illness, obstetric complications and duration of the illness on the effect of treatment with risperidone on psychopathological symptoms and deficits of working memory in patients with schizophrenia. The study involved 50 patients with schizophrenia, aged 16-50 years (mean 28 years), with duration of the illness 1-15 years (mean 4 years). The clinical and neuropsychological evaluation was performed before and after 3 months of risperidone treatment in dose 2-6 mg (mean 4.2 mg/day). Before treatment, a significantly higher intensity of psychopathological symptoms and of some cognitive dysfunctions was found in schizophrenic patients with family history of psychiatric illness and in those with a history of obstetric complications. After 3 months of risperidone treatment, no difference in the intensity of psychopathological symptoms or the performance on neuropsychological tests was observed among these groups. Patients with longer duration of the illness had less improvement in positive symptoms. The results of this study show that treatment with risperidone may lead to better compensation of psychopathology and of neuropsychological deficits originating in genetic predisposition and obstetric complications. Better effect of risperidone on psychotic symptoms is likely with the administration of the drug early in the course of the illness. PMID- 12647445 TI - [Opinions about long-term treatment of schizophrenia among psychiatrist in Gdansk region]. AB - Authors present opinions of 64 psychiatrists from Gdansk region working in university departments, in- and out-patients clinics. The aim of the study was to describe their opinion about selected problem in long-term treatment of schizophrenia. There were no significant differences between psychiatrists working in different centers. Psychiatrists had different opinions about long term treatment of schizophrenia, risk of relapse and side effects after antipsychotic drugs. Another opinions were quite similar to each other and were intact to present algorithms. PMID- 12647446 TI - [Self-efficacy and remission in schizophrenia]. AB - Self-efficacy is an important variable modifying ability to behavioral change, according to cognitive approach to mental disorders. It is a conviction of possessing psychobiological and cognitive abilities to establish desirable behavior. Self-efficacy determines efforts that are displayed in attempting to perform. The paper resolves self-efficacy of schizophrenic patients treated in hospital because of acute psychotic symptoms. Patients were examined twice: in the very beginning of hospitalization and after 4 weeks of neuroleptic treatment. Results reveal self-efficacy relation to negative, positive symptoms and global psychopathology in schizophrenia. If the therapy is based on neuroleptics, social self-efficacy predicts clinical improvement. The necessity of psychotherapy and rehabilitation focused on self concept aspects connected with competence to deal with social and life tasks is underlined. PMID- 12647447 TI - [Evaluation of depressive symptoms among schizophrenic patients during psychoeducation program]. AB - The article describes impact of psychoeducation on severity and frequency among schizophrenic patients. A group of 36 patients were evaluated using BDI, CDS and PANSS-G6 scales. There were no significant differences in severity of depressive symptoms before and after psychoeducation program. A slight fail in the frequency of depressive symptoms was observed after psycho-education. PMID- 12647448 TI - [Purpose in life among schizophrenic patients during psychoeducation program- preliminary report]. AB - The article describe impact of psychoeducation on purpose in life, will of life, hope, delight and satisfaction with life's attainments, relationship with family, health care and socio-economic situation among schizophrenic patients. There were no significant difference in on purpose in life hope and satisfaction with life's attainments, relationship with family, health care and socioeconomic situation. Patient after psychoeducation have significantly higher level of hope and delight. PMID- 12647449 TI - [The analysis of factors prior to positive symptoms relapse in schizophrenia]. AB - Schizophrenic relapses are an important problem for the sick persons and the society. They decrease the quality of patients' and their families life and also increase treatment costs. The aim of the study was to evaluate the type and frequency of the following factors during six months prior to relapse: patient non-compliance with pharmacotherapy, substance use, doctor's mistakes in antipsychotic treatment, absence of patient and family psychoeducation, stress situations, occurrence of physical disturbances and presence of additional psychiatric diagnosis. The studied group was composed of 60 patients with the diagnosis of paranoid schizophrenia (according to DSM-IV) that fulfilled the relapse criteria and were admitted in turn to psychiatric departments. Data were collected by specially constructed Questionnaire of Factors Prior to Schizophrenic Relapse. In the studied group the most frequent "risk factor" prior to relapse was patient non-compliance with pharmacological treatment (in 51.7% of subjects). PMID- 12647450 TI - [Prodromal symptoms of schizophrenics syndrome in children and adolescent]. AB - This presentation concentrates on modality and frequency analysis of prodromal schizophrenic symptoms of the children and adolescent. A sample (n-50) of children between 8 and 19 years was tested by structural interview. There were observed the signs resembling following symptoms: negative symptoms (74%), anxiety disorders (42%) and obsessive-compulsive disorders (20%). PMID- 12647451 TI - [The effect of sertraline on cognitive functions in patients with obsessive compulsive disorder]. AB - The aim of this study was to assess the effect of sertraline on psychopathological symptoms and cognitive functions and in patients with obsessive compulsive disorder (OCD). The investigated group consisted of 25 patients with OCD (12 male, 13 female) aged 17-47 (mean 29 +/- 9) years, duration of illness was 1-15 (mean 4 +/- 2) years. After treatment with sertraline, a significant improvement in OCD symptoms measured by YBOCS and in neuropsychological "frontal" tests were observed. Little correlation was found between the effect of sertraline on OCD symptoms and on cognitive dysfunctions. This may suggest that these two effects may be connected with different pharmacological properties of the drug. The effect of sertraline on OCD symptoms, similarly like other drugs inhibiting serotonin transporter (clomipramine, other SSRI) is associated with its influence on serotonergic system. On the other hand, the effect of sertraline on dopaminergic neurotransmission may be related to its favorable action on cognitive functions connected with the activity of frontal lobe. PMID- 12647452 TI - [Clinical efficacy of citalopram in patients with obsessive-compulsive disorder]. AB - 16 patients (9 female, 7 male; mean age 32.69 +/- 10.30) with coexisting obsessive-compulsive disorder (OCD) and depression were treated for 6 weeks with citalopram (10-60 mg; mean dosage after 6 weeks--25.71 +/- 12.83 mg). Their mental state was evaluated in the 7th, 14th, 28th and 42nd of treatment by the use of psychiatric examination and self-assessment clinical scales. After 6 weeks of treatment 5 patients were in full remission, 7 in partial remission (totally 75% of patients were responders) and 4 still suffered from OCD or depressive symptoms. In the 1st week of treatment 5 patients complained on temporary aggravation of symptoms but it turned out to have no prognostic value. It is recommended to find early signs of final clinical response. PMID- 12647454 TI - [The comparison of tianeptine and carbamazepine in benzodiazepines withdrawal symptoms]. AB - Dealing with benzodiazepine dependent creates as serious clinical problem that requires knowledge and experience. Abrupt discontinuation of benzodiazepines, particularly those with short half-life is not advised to avoid severe withdrawal syndrome. Reports from literature suggest use of carbamazepine and recently tianeptine as substances useful in treatment of benzodiazepine dependence. This paper presents a double-blind trial in which both, carbamazepine and tianeptine were used in treatment of benzodiazepiene withdrawal syndrome. Patient mental state was evaluated by using questionnaire SCL-90, Beck Depression Inventory and specifically designed questionnaire assessing severity of symptoms following benzodiazepine withdrawal. It appears from this study that both drugs (carbamazepine and tianeptine) are comparable, safe and efficient in treating benzodiazepine withdrawal symptoms. PMID- 12647453 TI - [Carbamazepine in the treatment in psychiatric disorders based on retrospective clinical study]. AB - Medical records of patient treated in I Department of Psychiatry Medical University of Gdansk in 1996-2000 were analysed. Carbamazepine was used in 383 patients, 209 male and 174 female. Mean age for the whole group 41.2 years, for male 40.6, for female 41.9. Carbamazepine was mainly used in affective disorders (F30-33) and organic affective disorders (F06.3)-107 persons. Second the most common indication was organic personality disturbances (F07.0)-121 persons. Carbamazepine was used in 100-1200 mg per day (mean dose 415 mg) during 5-189 days (mean 36 days). Treatment was well tolerated. Only 24 patients (6.3%) have mild side effects. Drug level in blood was checked in 19 patients during two last years of analysed material. Carbamazepine was used mainly with tricyclic antidepressants (30.3%), second generation antidepressive drugs (22.2%) and typical neuroleptics (11.1%). More than half of patients (55.6%) were recovered. Based on clinical material carbamazepine seems to be active agent in treatment of affective disorders and diseases connected with organic brain lesion. PMID- 12647455 TI - [Therapeutic possibilities regarding somatoform disorders in soldiers]. AB - The authors used the ICD-10 classification and presented the most common psychopathological syndromes from the category "neurotic disorders connected to stress and somatoform disorders" present amongst recruited soldiers. Recruitment to the army is a "situation which is endangering to the mental health" (from the Act on mental health protection), however socio-political changes that took place in our country are an additional factor which can lead to such disorders. The limitations in treatment of these disorders, which are due to army specifications, cause for the majority of those soldiers affected to be unable to carry out their duty. As a consequence of this, civil units carry out the therapeutic processes. Particular attention, amongst the other disorders described, is attributed to the so-called organ neuroses. The authors stress upon the need for psychoprophylactic actions which would reduce the health consequences of being recruited into the army. PMID- 12647456 TI - [Follow-up study of quality of life and treatment of eating disorder: dynamics of the depressive and anxiety symptoms]. AB - A group of 56 female patients with eating disorders (36 anorexic and 20 bulimic) aged 16-48 (average 23.9) was studied. According to DSM-IV and ICD-10 among anorectic 8 (14.3%) represented restricting type and 28 (50%)-bulimic type, among bulimic-12 (21.4%)-purging type and 8 (14.3%)-nonpurging type. They were treated average 21.5 months in the I Department of Psychiatry Medical University of Gdansk (some of them were outpatients) between 1995-2000. Duration of disorders was 4 months-10 years (X +/- SD33.8 +/- 28.6 months). Follow-up period lasted from 4 months to 2 years. All patients were examined twice time (before and after treatment) with the following methods: Structured Questionnaire, Hospital Depression and Anxiety Scale Beck Depression Inventory (BDI), PIL by Crumbaugh and Maholick, Spielberger Self-Evaluation Questionnaire. The aim of our examination was to define changes in the state of patients after complex treatment (pharmacotherapy and psychotherapy) and important parameters of quality of life in our groups. After complex treatment (pharmacotherapy, individual and group psychotherapy) which lasted average 21.5 months we observed significant improvement of depressive and anxiety symptoms. Pharmacotherapy included tricyclic antidepressants (55.4%), tetracyclic antidepressants (10.7%), SSRI (44.6%), neuroleptics (10.7%), carbamazepine (32.1%). Some patients were treated more than one medicine. We received high correlation between ideal weight and weight after treatment. Therefore we supposed that our complex therapy was effective and improved approval of self-image and weight. Significance of life after treatment depended on intensity of depression in BDI and intensity of anxiety as state. The long lasting complex therapy significantly lowered depressive and anxiety symptoms (in subjective and objective aspects) in our group of patients with eating disorders. PMID- 12647457 TI - [Quality of life in patients with irritable bowel syndrome]. AB - The authors have evaluated the quality of life of 120 persons in three equal groups (those with irritable bowel syndrome, those with neurosis but no psychosomatic symptoms and healthy individuals). The Scale of Quality of Life constructed on the basis of the Rotterdam Symptom List, was applied. The authors have also observed the changes that took place in the noted quality of life in the course of therapy. The major conclusion that arises from this study is the following: in the case of irritable bowel syndrome, psychotherapy should be used besides typical pharmacological treatment. PMID- 12647458 TI - [The analyses of psychiatric treatment among a group of people applying for disability pension benefits]. AB - The study includes data of 109 people who were examined by psychiatrists and psychologists of their pension demands concerning statement for the Regional Court in Lublin, in 1999-2001 period. The period of treatment and the frequency of calls to PZP have been taken into consideration. The number of hospitalizations has also been taken into account. Then, pharmacological treatment was examined. Some discrepancies have been found among algorithms of pharmacotherapy and the practice of treatment. Insufficient and inconsistent application of psychotropic medicine has been proved. It turned out that 1/3 of men and 1/5 of women of the examined group weren't patients of PZP. They were treated by physician (family doctors) or neurologists. PMID- 12647459 TI - [Dependence on benzodiazepine and treatment of withdrawal syndrome]. AB - The 45 patients long-term benzodiazepine users were analyzed because of the diagnosis, the purpose of benzodiazepine onset, the management of benzodiazepine discontinuation and katamnestic examination. Most of the patients suffered from anxiety disorders or mixed anxiety-depressive disorders and they could be successfully with antidepressive drugs treated. Benzodiazepines were also often used without any clinical reason. PMID- 12647461 TI - ["European Service Mapping Schedule" (ESMS)--instrument used to describe the structure of mental health care]. AB - There is many changes in psychiatric service last years. In deinstitutionalization process arising new diversified and wide integrated psychiatric services. Traditional names lose significance for example: open or stationary care. Moreover the same names, have different meaning in different centers or countries. Because of that processes in year 1997 EPCAT group with funding from European Commission Biomed grant developed ESMS instrument by Sonia Johnson and Robert Kuhlmann, to modern description and classification of psychiatric services. Instrument is based on standardizing classification accessible services, with different care models in Europe. In this work authors presented structure of questionnaire and typical applications in catchment areas. That compatible with European standards of psychiatric service instrument, portray gaps in old standards system and indicate developmental trends new psychiatric health service in medical care. Instrument ESMS indicates another possibilities in service study area for example analysis and comparison individual service components and helps in planning local development of psychiatric care services. PMID- 12647460 TI - [Treatment in psychiatric day hospital in comparison with inpatient wards in different European health care systems--objectives of EDEN project]. AB - The paper presents the objectives and design of an ongoing multicenter randomized, controlled trial EDEN (European Day Hospital EvaluatioN). The EDEN study aims to evaluate the efficacy of acute psychiatric treatment in a day hospital setting in five European centres: Dresden, London, Michalovce, Prague and Wroclaw. The main hypothesis is that day hospital treatment for acute psychiatric patients is as effective as conventional inpatient hospital care. The objectives of the study are to evaluate the viability and effectiveness of day hospitals for acute psychiatric treatment, to identify subgroups of patients with a more or less favourable outcome so that the treatment setting might be specifically applied and to ascertain the cost-effectiveness of day hospital treatment compared to conventional inpatient treatment. The study utilises a Randomised Controlled Trial (RCT) design with repeated measures at a maximum of six time points: at admission (t1), one week after admission (t2), four weeks after admission (t3), discharge (t4), three months after discharge (t5), and 12 months after discharge (t6). A combination of well-established standardised assessment instruments and open questions is used in 6 time periods. If the findings accept the main hypothesis of the study, some practical consequences could be inevitable: at a mental health policy level, these results could lead to an increase in the capacity of day hospitals; at the clinical level clinicians could redefine their concepts of care to consider the day hospital as an alternative to conventional inpatient treatment; from economic point of view could lead to reduction of treatment costs. PMID- 12647462 TI - [Comparison between a SCAN diagnosis and a clinical diagnosis]. AB - The aim of this paper is to present comparison of clinical diagnoses and SCAN (Schedules for Clinical Assessment in Neuropsychiatry, version 2.1) diagnoses of patients treated in day care and inpatients wards. SCAN was translated at Medical University of Wroclaw according to WHO permission. It is a part of multicenter study evaluating effectiveness of treatment in conventional psychiatric hospital and day hospital for acute psychiatric patients. Clinical diagnoses were made using ICD-10 classification system. Till now 56 patients were examined, 15 men and 39 women in age from 21 to 65 years old, median age 45 years old. Among them 31 patients were treated in day care wards, and 25 patients were treated in inpatients wards. During standard medical procedure diagnoses of 25 patients were classified into the block F2, and 19 patients into the block F3 of ICD-10 classification. Schizophrenia were diagnosed in 16 patients of, 19 patients had diagnosis of depressive disorder, and 21--of other mental disorders. Clinical diagnoses were in most of the cases single one; there were only 6 cases with double (comorbid) diagnoses. SCAN diagnoses were multiple, ranged from 1 to 12 (median 3) for one patient. Many concomitant SCAN diagnoses referred to tobacco use, sleep disorders, and somatoform disorders. Cohen's kappa coefficient for the block F2 was 0.78, for schizophrenia--0.62, for the block F3--0.42, and for depression--0.36. Results showed low agreement between clinical and SCAN diagnoses, particularly for mood disorders. The explanations of such discrepancies could be: no strict compliance with rules of ICD-10 by clinicians and avoidance of comorbid diagnoses by clinicians or errors in the SCAN coding algorithms. Further analyses are needed to characterize more precisely the sources of discrepancies. PMID- 12647463 TI - [The Groningen Social Disabilities Schedule (GSDS) in the multicenter study EDEN]. AB - The paper presents an instrument for estimation of social functioning Groningen Social Disabilities Schedule (GSDS). Analysis of reliability in patients' social disability assessment made by research assistants during the GSDS training in research project EDEN was also described. PMID- 12647464 TI - [Presentation of the cost-effectiveness technique questionnaire--the possibility of the Polish adaptation]. AB - An increase of interest of cost-effectiveness begins from 1999 when act about general insurance was in force. The same situation was observed a few years ago in Europe. It was the reason why created and standardized questionnaire: European Service of Mapping Schedule--(ESMS)--for describing mental health system, European Socio--Demographic Schedule--(ESDS)--for presentation sociodemographic situation, The International Classification of Mental Health Care--(ICMHC)- questionnaire used to assess therapeutic process and Client Service Receipt Inventory--(CSRI)--for estimate cost-effectiveness. These questionnaires are used in clinical trial "European of Day Hospital Evaluation" conduct in Wroclaw. PMID- 12647465 TI - News media reporting on civil litigation and its influence on civil justice decision making. AB - The news media have the potential to act as a powerful influence on the civil litigation system, influencing decision making in particular cases and on the system more generally as media reports influence the decision making of various participants in the system. This paper reviews the research that has examined the relationship between news media reporting and civil litigation and proposes a framework that integrates this work and provides guidance for future research efforts. Specifically, we discuss the nature of media reporting on civil litigation, perceptions of the civil litigation system held by the public and legal actors, and the potential influence of news reporting about civil litigation on the decision making of jurors, judges, civil litigants, and policymakers. Overall, the research suggests that news reporting of civil litigation presents a systematically distorted picture of civil litigation and that this reporting can influence perceptions and outcomes of civil litigation in various ways. However, there are many gaps in the existing research that need to be filled. The proposed organizational scheme helps to identify ways that future research can provide links between the findings of existing research and to identify ways in which this research can be extended to new areas. PMID- 12647466 TI - Sexual harassment stories: testing a story-mediated model of juror decision making in civil litigation. AB - The story model of juror decision-making proposes that jurors use personal experience and information presented at trial to create stories that guide their verdicts. This model has received strong empirical support in studies using criminal cases. The research presented here extends the story model to civil litigation and tests a story-mediated model against an unmediated model of jury decision-making. In Phase 1, content analysis of mock juror responses to 4 realistic sexual harassment cases revealed prototypic plaintiff and defense stories. In Phase 2, these prototypic stories were included as mediators in a model predicting verdicts in 4 additional sexual harassment cases. Mock juror attitudes, experiences, and demographics were assessed, then attorneys presented abbreviated versions of 4 actual sexual harassment cases. Path analyses provided support for the story-mediated model, which added significantly to the amount of variance accounted for in the outcome measures of verdict, commitment to verdict, and confidence times verdict. Implications for sexual harassment and other types of civil cases are discussed. PMID- 12647467 TI - Measuring damages for lost enjoyment of life: the view from the bench and the jury box. AB - Civil jury instructions are inconsistent in defining what constitutes noneconomic damages, which may include pain, suffering, disability, disfigurement, and loss of enjoyment of life (LEL), among other injury sequelae. This inconsistency has been manifested recently in court decisions that have considered whether LEL should be treated as a separate element of noneconomic damages, distinct from pain and suffering. This paper reviews the case law on this issue and also describes a jury simulation experiment. Mock jurors awarded damages after they received instructions on noneconomic damages in which LEL was (1) not identified as a distinct element of damages; (2) defined as an element of damages distinct from pain and suffering, but participants awarded a single amount for noneconomic damages; or (3) defined as a distinct element of damages, and participants awarded separate amounts for LEL and pain and suffering. Instructions about LEL resulted in larger awards, but only when mock jurors also made a separate award for that element of damages. PMID- 12647468 TI - Here comes the judge: the influence of judge personal characteristics on federal sexual harassment case outcomes. AB - This study explored the effects of judges' personal characteristics (gender, race, age, and political affiliation) and case characteristics on the outcomes of federal cases of hostile environment sexual harassment. Results revealed that even after controlling for the effects of relevant case characteristics (e.g., severity of the harassment), judges' personal characteristics influenced case outcomes. Specifically, younger judges and Democrat judges were more likely to find for the plaintiff (the alleged victim of harassment). The probability that the decision would favor the plaintiff was only 16% when the case was heard by an older judge but 45% when heard by a younger judge. The probability that the decision would favor the plaintiff was only 18% when the case was heard by a judge who had been appointed by a Republican president but 46% when the judge had been appointed by a Democrat president. PMID- 12647470 TI - New visual technologies in court: directions for research. AB - Litigation is being transformed by new visual communication technologies, including videoconferencing, PowerPoint, and computer animations. Yet the effects of these visual technologies on legal decision making are largely unknown. In order to understand better the most pressing issues surrounding technology in the courtroom, psychologists, lawyers, and representatives from technology companies and funding agencies attended a Research Conference on Courtroom Technology organized by the Federal Judicial Center. The goals of the conference were to identify issues raised by courtroom uses of new technologies that could be illuminated by empirical research and to suggest designs and methods for conducting that research. This paper emerged from that conference. The authors provide an overview of considerations that should guide research in this area, including a framework that takes into account features of the technology, the audience, and the legal strategy of the user of the technology. They outline a paradigm for conducting such research, illustrate it with several possible empirical studies of varying levels of experimental and conceptual complexity, and identify directions for subsequent research. PMID- 12647469 TI - Organizational downsizing and age discrimination litigation: the influence of personnel practices and statistical evidence on litigation outcomes. AB - The present study examined relationships between reduction-in-force (RIF) personnel practices, presentation of statistical evidence, and litigation outcomes. Policy capturing methods were utilized to analyze the components of 115 federal district court opinions involving age discrimination disparate treatment allegations and organizational downsizing. Univariate analyses revealed meaningful links between RIF personnel practices, use of statistical evidence, and judicial verdict. The defendant organization was awarded summary judgment in 73% of the claims included in the study. Judicial decisions in favor of the defendant organization were found to be significantly related to such variables as formal performance appraisal systems, termination decision review within the organization, methods of employee assessment and selection for termination, and the presence of a concrete layoff policy. The use of statistical evidence in ADEA disparate treatment litigation was investigated and found to be a potentially persuasive type of indirect evidence. Legal, personnel, and evidentiary ramifications are reviewed, and a framework of downsizing mechanics emphasizing legal defensibility is presented. PMID- 12647471 TI - Postmenopausal hormone therapy in the aftermath of the WHI. What patients need to know. AB - HT is now relegated primarily to the treatment of menopausal symptoms. It should be used for this indication at the lowest effective dose and for the shortest duration possible. Consideration should be given to other strategies to control symptoms when patients are willing to try less effective but potentially safer alternatives to HT. There is little support for the HT to prevent chronic conditions. The risks of combined HT appear to outweigh the benefits. However, the jury is still out regarding the use of estrogen alone. Ongoing research is likely to provide more informaton about the effects of HT on quality of life, cognition, and prevention of Alzheimer's disease. These potential benefits could tip the balance in favour or HT use for some patients. Any decision about starting or stopping HT requires a comprehensive discussion between the physician and the patient regarding benefits, risks, and uncertainties. The WHI results underscore the importance fo well-designed randomized controlled trials to inform our treatment discussions. Even the best observational studies have the portential to mislead us. PMID- 12647472 TI - The good news about bad news. PMID- 12647473 TI - Update on therapy for acute and chronic heart failure. Applying advances in outpatient management. AB - Care of patients with heart failure is often challenging and involves proper therapeutic selection from the pharmacologic, interventional, and mechanical options available. In this article, Drs Almeda and Hollenberg discuss the latest approaches and guidelines to treatment of acute and chronic heart failure in patients with left ventricular systolic dysfunction. They review the benefits, risks, and myriad considerations that physicians need to assess before making choices about best medical management. PMID- 12647475 TI - Titration of beta-blockers in heart failure. How to maximize benefit while minimizing adverse events. AB - Data from large clinical trials indicate that beta-blocker therapy can be successfully initiated and adjusted upward in most patients with stable chronic heart failure who already take standard heart failure therapy. Such therapy typically includes ACE inhibitors, diuretics, and digoxin. With optimal titration and maintenance strategies, beta-blockers are effective and well tolerated in these patients. It is recommened that all patients with clinically stable mild to moderate chronic heart failure (NYHA class II or III), no contraindications to beta-blocker use, and an LVEF less than 40% should be treated with beta-blockers. Based on the results of recent clinical trials on heart failure, beta-blocker therapy should be initiated at a low dose and slowly tirtrated upward as tolerated. A patient's heart failure should be stable for at least 2 weeks before the dose is adjusted upward. Slow titration facilitates maximal tolerability. In primary care practice, physicians should apply titration strategies and target dosed that have been demonstrated to reduce morbidity and mortality in clinical trials. Although worsening heart failure or other adverse events occur in a minority of patients who take beta-blockers, these effects can be managed by adjusting the dose of ACE inhibitor or diuretic, or both, or by temporarily withholding the beta-clocker. Currently, professional treatment guidelines recommend beta-blocker therapy in combination with ACE inhibitors an diuretics as the standard of care in the treatment of heart failure. PMID- 12647474 TI - Improving outcomes in diastolic heart failure. Techniques to evaluate underlying causes and target therapy. AB - Abnormal diastolic function is a common cause of clinical heart failure, particularly among elderly patients. Through early diagnosis and careful management of diastolic dysfunction, these patients can expect improved functional capacity and, in some cases, a favorable long-term outcome. In this article, Drs Torosoff and Philbin discuss how to confirm the diagnosis of diastolic heart failure through objective testing. Current approaches to the treatment of symptoms, including reduction of intravascular volume, heart rate control, and elimination of precipitating factors, are also presented. PMID- 12647476 TI - A curious case of ascites. Chylous ascites caused by Kaposi's sarcoma. PMID- 12647477 TI - Aspiration pneumonia. Recognizing and managing a potentially growing disorder. AB - Gross aspiration of liquid or particulate matter into the lung can result in severe hypoxemia, pulmonary infiltrates in dependent lung regions, fever, and leukocytosis. The initial lung injury is primarily due to inflammatory mediators rather than infection. The responsible bacterial pathogens differ between community-acquired and nosocomial aspiration pneumonia. Many aspiration pneumonias are mixed aerobic-anaerobic infections. Enteric gram-negative bacilli and S aureus are more common in nosocomial aspiration pneumonia. Current treatment guidelines support initial empirical antibiotic therapy in patients with severe aspiration pneumonia pending culture results. Appropriate initial treatment improves outcome. Antimicrobial therapy for aspiration pneumonia is often empirical and should be based on patient characteristics, the setting in which aspiration occurred, the severity of pneumonia, and available information regarding local pathogens and resistance patterns. PMID- 12647479 TI - Patient notes. Asthma. PMID- 12647478 TI - Cost-effective workup for tonsillitis. Testing, treatment, and potential complications. AB - Despite increased strictness in surgical criteria, tonsillectomy continues to be one of the most common outpatient surgical procedures performed in the United States. The primary care physician is integrally involved in the diagnosis and treatment of tonsillitis, the chief reason for tonsillectomy. This article gives guidelines for diagnosis and management of tonsillitis and provides an overview of its potential complications. PMID- 12647480 TI - [The clinical significance of postictal electrocardiographic changes mimicking acute myocardial infarction]. AB - The electrocardiogram (ECG) is of critical importance in the diagnosis of acute myocardial infarction. Clinical conditions such as acute pericarditis, esophageal rupture, pancreatitis, subarachnoid hemorrhage, perforated duodenal ulcer, pneumothorax and status following elective DC cardioversion result in ECG changes that include ST elevation and T wave inversion. This report aims to increase the awareness of non-cardiac syndromes, with ECG abnormalities mimicking acute myocardial infarction, and thus to avoid unjustified thrombolytic therapy. We describe the case of a patient after epileptic seizures and pathological EEG pattern. The ECG showed repolarization abnormalities suggestive of evolving acute myocardial infarction. The cardiac enzymes (except normal Troponin I) were severely elevated and coronary angiography was normal. PMID- 12647481 TI - [Evaluation of the impact of a clinical pharmacist on the quality and cost of pharmacotherapy in a general surgical department, the Hillel-Yaffe Medical Center]. AB - BACKGROUND: Clinical pharmacy could be defined as an umbrella of services aimed at maximizing the pharmacotherapeutic effect, minimizing the risk for developing adverse drug effects and reducing drug expenditures for the health care system. Clinical pharmacists, in Israel and abroad, have been practicing in different health care settings aiming to implement the principles mentioned above. Most of the articles previously published reviewed the role of the clinical pharmacist in internal medicine wards or in intensive care units. This article focuses on the role of the clinical pharmacist in a general surgery department and the influence of this service on the quality and cost of pharmacotherapy and on intravenous antibiotic therapy in particular. AIMS: Improving the quality of pharmacotherapy while optimally using economic resources in a general surgery ward. METHODS: During September 1999 to August 2000, the clinical pharmacist joined physician rounds in the surgical ward. During the rounds he advised physicians about clinical and economical aspects of drug treatment, and collected data on his activity on a specific designated form. The data was processed on a central computerized database, and analyzed to determine the influence of the clinical pharmacist on clinical and economical outcomes. RESULTS: At the end of the study period the following were found: The pharmacist made 219 interventions. These included adverse drug effects, which were identified and/or prevented. When considering the economic data, a substantial reduction of 56% in intravenous antibiotic therapy was noted (direct saving of about 140,000 NIS). Such savings were accompanied by changes in the prescribing patterns in the department, which were translated into increased rate of oral antibiotic prescribing and reduction in the use of certain i.v. antibiotics while undamaging the quality of the pharmacotherapeutic effect. CONCLUSION: The results of this study indicate that participation of a clinical pharmacist during physicians rounds improved the quality of the pharmacotherapy, assisted in changing clinicians prescribing habits, and at the same time significantly reduced the direct expenditures on medications in general and of intravenous antibiotic therapy in particular. PMID- 12647482 TI - [The diverse etiology of hiccups]. AB - Hiccup is a sudden contraction of the inspiratory muscles, followed by an abrupt closure of the glottis, thus producing a characteristic sound. Hiccups serve no known physiologic function. Mostly, it is a benign symptom which terminates without treatment. Persistent and intractable hiccups may indicate an organic disorder, thereby requiring evaluation based on history, physical examination, and selected laboratory tests. The treatment is based on the organic disorder, if it is found. The combination therapy with cisapride, omeprazole and baclofen is the most effective empiric treatment. We describe a number of case reports of patients with hiccups due to digitalis intoxication, ischemia of the inferior wall of the heart, peptic disease and gastroesophageal reflux disease. PMID- 12647483 TI - [Undiagnosed celiac disease in adults unmasked by endoscopy]. AB - BACKGROUND: Celiac disease may be present in many patterns other than the classical manifestations of diarrhea and malabsorption. The prevalence of celiac disease is greater than usually reported as mild cases are often undiagnosed. Diagnosing celiac disease may have major health significance. AIMS: The aim of our study was to determine the prevalence of celiac disease in adults appearing for endoscopy with abdominal pain, in an open access setting. METHODS: Two hundred and seventy consecutive, non-selected adult patients undergoing upper endoscopy for abdominal pain were included in the study. Three biopsies were routinely taken from the second part of the duodenum. In those cases with flattened villi on histology, antiendomysial antibody (AEA) was tested. RESULTS: The prevalence of celiac disease among Israeli patients undergoing endoscopy for abdominal pain was high (1:23), greater than its highest reported prevalence in general populations (1:200-300). CONCLUSIONS: Routine small bowel biopsies should be considered in patients undergoing upper endoscopy for otherwise unexplained pain. PMID- 12647484 TI - [Prevalence and characteristics of myocardial infarction, diabetes and hypertension in the adult Jewish population: results from the Negev Nutritional Study]. AB - BACKGROUND: Monitoring the prevalence of chronic conditions such as chronic heart disease, diabetes and hypertension in adult populations is essential for health services planning and identification of populations at high risk. OBJECTIVES: To describe the prevalence of self-reported conditions such as myocardial infarction, diabetes mellitus and hypertension in the Jewish Negev population and the patterns of use of health services and dietary behavior of persons suffering from these conditions. METHODS: A random proportional geographic cluster sample of the adult Jewish population (n = 1159, age 35+) from the Negev area was interviewed at home between 1998 and 1999. The interview included questions regarding chronic conditions, patterns of health services use and dietary behavior. RESULTS: Men had twice the prevalence of myocardial infarction and underwent more invasive cardiac procedures than women. The highest prevalence of myocardial infarction and hypertension were found in Central- and Eastern European-born persons while the highest prevalence of diabetes was found in Western-born participants. Of the participants < 61 years of age, 19% reporting diabetes and 33% reporting hypertension did not use medication and were not adhering to an appropriate diet. Thus, one-third of those with reported hypertension and 15% of those reported as diabetics were not adhering to any treatment. The prevalence reported in this study was higher than the national data. CONCLUSIONS: The data collected showed a higher prevalence of chronic diseases among the southern Israeli population as compared with the national data. Among people with chronic diseases, high percentages are not treated. The information reported here may help in the allocation of health services for the south of Israel and in the identification of populations at risk. PMID- 12647485 TI - [Comparison between colonic J-pouch anal anastomosis and straight coloanal anastomosis following low anterior resection of the rectum]. AB - BACKGROUND: Low anterior resection, colo-anal anastomosis with anal sphincters preservation has become the routine procedure for the treatment of low rectal cancer. This method avoids the need for a permanent colostomy without compromising oncological principles. However, many of these patients experience symptoms of fecal frequency, urgency and variable degree of incontinence. Improvement with the functional results has been reported in patients in whom continuity has been restored by means of colonic j pouch. AIM: To compare the outcome of anterior resection and colonic pouch anal anastomosis (CPAA) with the conventional straight coloanal anastomosis (SCAA). MATERIAL & METHODS: The files of patients who underwent anterior resection with CPAA or SCAA between 1994-2001 were analyzed retrospectively. Patients filled out an updated questionnaire regarding their functional outcome. RESULTS: Thirty nine patients underwent CPAA and 42 SCAA. Mean follow-up was 31 months. There was no perioperative mortality and the rate of complications was similar in the two groups. Bowel movements per 24 hours were 4.2 and 6 in the CPAA and the SCCA groups respectively. The bowel movements during night were 0.4 and 1.5 respectively. Sixty percent of patients after CPAA had 3 or less bowel movements per 24 hours as compared to 44% of patients after SCAA. Continence scores were 3.9 and 3.5 in patients after CPAA and SCAA respectively. Seventy two percent of patients after CPAA reported continence scores of 4 or more as compared to only 51% of patients in the SCAA group. CONCLUSION: Our results confirm previous reports that anterior resection and CPAA improves the functional outcome and quality of life after sphincter saving operation for low rectal cancer. PMID- 12647486 TI - [Monitoring heart failure with brain natriuretic peptide]. AB - Brain natriuretic peptide (BNP), is a 32 amino acid containing cardiac hormone released from the ventricles due to increased filling pressure and left ventricle wall stretch. For over a decade since its discovery, information is mounting to suggest its superiority over the other natriureic peptide, ANP, as a reliable measure of left ventricular systolic dysfunction. Measurement of BNP concentration in blood emerges as a sensitive and specific test for the identification of patients with congestive heart failure in the urgent-care setting. It is a potent, cost-effective addition to the diagnostic armamentarium and helps in the titration of following therapy and prognosis evaluation. PMID- 12647487 TI - [The effectiveness of estrogen replacement therapy to reduce ischemic heart disease--revaluation]. AB - Cardiovascular disease is the leading cause of death of postmenopausal women in developed countries. One of the main reasons for providing hormone treatment for women was the possible prevention of ischemic heart disease (IHD). There are many cellular and molecular studies showing the effectiveness of estrogen in reducing risk factors of IHD, but none of these studies deal directly with the reduction of IHD. However, the studies that "prove" the direct effectiveness of estrogen in primary prevention of IHD are only observational studies. Doubts were raised, when two prospective, randomized, blinded studies (HERS and ERAT) were published, refuting the beneficial effect of estrogen in secondary prevention of IHD. Last July, part of the WHI study was published, showing increased risk for ischemic heart disease in patients treated with continuous combined HRT. In this era of Evidence Based Medicine, it is necessary to support decisions of treatment with prospective, randomized, blind trials. The publication of HERS I and II, ERAT and WHI, raised doubts concerning the ability of estrogen replacement treatment to be considered as primary or secondary prevention of IHD. PMID- 12647488 TI - [Out of hospital deliveries: incidence, obstetrical characteristics and perinatal outcome]. AB - Traditionally, women used to deliver their babies at home. In 1927, in England and Wales, 85% of births took place at home. By 1970 the position was reversed. The move from home to institutional delivery has been accompanied by changes in the institutions themselves and in the type of care provider. There are two kinds of out-of-hospital deliveries: 1. Planned home deliveries--women who decide to deliver in their home with the assistance of midwives or other consultant obstetric facilities. Few cases from this group, however, end up in the hospital; 2. Unplanned home deliveries or delivery en-route to the hospital--when women enter the active phase of labor rapidly, resulting in accidental out-of-hospital deliveries. The study aims to review the available literature and to describe the incidence, obstetrical characteristics and perinatal outcome of out-of-hospital deliveries. PMID- 12647489 TI - [Ptosis--etiology, diagnosis and management]. AB - Ptosis is a common complaint in patients who refer to ophthalmologists especially oculoplastic surgeons. Occasionally, ptosis may be the presenting sign or symptom of a systemic disease. Therefore, it is of great importance to know the causes of ptosis, its clinical presentation, and the systemic evaluation that one must undergo before going into surgery. This review presents the common causes of ptosis according to age of appearance, systemic diseases that must be ruled out, and the surgical options that exist. PMID- 12647490 TI - [Toxoplasma and toxoplasmosis]. AB - Toxoplasmosis is a zoonotic protozoal disease, caused by an obligatory intracellular parasite of the genus Toxoplasma. The disease is widely distributed affecting more than a billion milliard people, worldwide. Raising sheep and cattle, handling and eating raw meat, interaction with domestic cats and climate conditions play an important role in the distribution of the disease. Both Toxoplasma and Plasmodium belong to the Phylum Apicomplexa, and are, therefore, almost similarly sensitive to anti-malarial drugs. Toxoplasmosis is considered to be viscerotropic in adults and children and neurotropic in fetal and newborn children. An accurate and fast diagnosis of the disease is highly important, particularly in pregnant women, since the results may affect both the mother and her fetus. The present study summarizes the old and new techniques available and their importance in the diagnosis of the disease. PMID- 12647492 TI - [Hyperemesis gravidarum: an updated review]. AB - Hyperemesis gravidarum is a common entity characterized by severe and protracted vomiting during pregnancy not attributed to any organic or metabolic condition other than pregnancy itself. It may lead to fluid and electrolyte imbalances as well as ketosis, and its diagnosis is made per exclusion. While the accurate etiology and pathophysiology of hyperemesis gravidarum remain largely unknown, numerous theories have been proposed in attempts to explain this peculiar phenomenon. The different therapeutic modalities aim to correct the hypovolemic state, electrolyte imbalance and ketosis. The present manuscript reviews the pertinent medical literature regarding this illness in order to refine our understanding of its pathophysiology, clinical manifestations and therapeutic modalities. PMID- 12647491 TI - [Oral anticoagulation treatment--control, risks, benefits, and informed consent]. AB - The most common indications for warfarin therapy are atrial fibrillation, the presence of a mechanical heart valve and venous thromboembolism. The risks and benefits of treatment for these conditions have been fairly well characterized and recent consensus groups have provided guidelines which are nearly identical. Although we can define the recommended range of the PT in INR units, it is often difficult under the best of circumstances to keep the INR in the desired range. Physicians often dont recognize the limitations of INR testing. Furthermore, the continued risk of thromboembolic and bleeding complications despite state of the art treatment might not be recognized by the patient. This places the physician at risk of a malpractice suit and emphasizes the need for informed consent. The following review will summarize current recommendations for the major conditions requiring long-term management with oral anticoagulants, identify problems with PT-INR testing, consider when the laboratory should contact the physician urgently, define acceptable control of the PT-INR during audits, and finally suggest information for the patient, which might be used in an informed consent form. PMID- 12647493 TI - [The medical treatment of glaucoma]. AB - Chronic open angle glaucoma is a one of the leading causes of irreversible blindness. Elevated intraocular pressure is a major risk factor for the progression of this disease. At present, most patients suffering from open angle glaucoma have started medical therapy. The goal is to reduce the intraocular pressure to an individualized target pressure in an effort to delay the progression of damage to the optic nerve. Until a decade ago, topical beta adrenergic antagonists, adrenergic agonists, miotics and oral carbonic anhydrase inhibitors comprised the common medications in use. In the past decade many new drugs have been introduced. These drugs exert less systemic side effects and are very effective in lowering the intraocular pressure and furthermore, are easier to comply with. These include: local carbonic anhydrase inhibitors, prostaglandin F2a analogues and a2 adrenergic selective agonists. Due to lack of consensus as to the initial medication of choice for the commencement of treatment, the traditional tendency to initiate treatment with a local beta adrenergic antagonist persists. This review attempts to familiarize the reader with the new arsenal of glaucoma medications. PMID- 12647494 TI - [Human transmissible spongiform encephalopathies]. AB - A new variant of Creutzfeldt-Jakob disease was discovered in 1996. This new variant was found to be related to encephalopathy. There are 5 known human transmissible spongiform encephalopathies, all created by prions. The prion, a pure protein, smaller than a virus is derived from a normal glycoprotein of human cells wall. The function of this protein is unknown but supposed to be connected with copper incorporation into superoxide dismutase. Creutzfeldt-Jakob disease is an early dementive disease with loss of neurons, accumulation of amyloid and spongiform degeneration of the brain. We discuss the epidemiology, diagnosis and therapy of Creutzfeldt-Jakob and its variant. The increased life expectancy will certainly cause these diseases to be more common, thereby having a definite affect on the economy in both developing and developed countries. Future therapy will concentrate on either preventing the creation of pathologic prions or extracting intracellular prions. PMID- 12647495 TI - [Intestinal diseases in the City of David]. PMID- 12647496 TI - Gastroparesis and the gastric pacemaker: a revolutionary treatment for an old disease. AB - Gastroparesis--or delayed gastric emptying--can apply to either solids or liquids but the term (gastroparesis) usually refers to delayed solid emptying, in an appropriate clinical setting. Impaired gastric emptying can occur acutely in a variety of conditions from neurologic to metabolic. Intractable nausea and vomiting, and the inability to tolerate oral intake, occurs frequently in patients with severe gastroparesis. The patients' nutritional status is adversely affected by gastroparesis as are the socioeconomic aspects of patient's lives. The previous treatments of gastroparesis, such as medications and surgery, were often not successful. Our recent understanding of the normal gastric electro mechanical function and its abnormalities/dysrythmias has led to the development of a gastric electrical stimulator, analagous to other devices used to stimulate dysfunctional organs. Gastric electrical stimulation is associated with improvements in symptomatic relief, nutritional status, health resource utilization and costs and is now approved by the FDA as a humanitarian use device. It offers a new and hopeful therapy for patients with refractory gastroparesis where none have existed before. PMID- 12647497 TI - A recovering physician being monitored by the Mississippi Recovering Physicians Program and prescribed Ultram. PMID- 12647498 TI - If these walls could only talk. PMID- 12647499 TI - The treatment of pneumonias in 1920. PMID- 12647500 TI - Shackleton's cardiac phobia. PMID- 12647501 TI - Introduction from guest editor, Dr. William Cleveland. PMID- 12647502 TI - A tribute to Dr. Louis W. Sullivan. PMID- 12647503 TI - Introduction from President George H. W. Bush and Mrs. Barbara Bush. PMID- 12647505 TI - Louis W. Sullivan, M.D. Secretary of the U.S. Department of Health and Human Services, 1989-1993. PMID- 12647504 TI - Knowledge Wisdom Excellence Service. PMID- 12647506 TI - Morehouse School of Medicine: the years after health and human services. PMID- 12647507 TI - The legacy of leadership. PMID- 12647508 TI - Practice transitions--is your practice ready for your retirement? PMID- 12647509 TI - Systematic review of the current literature related to disability and chronic fatigue syndrome. PMID- 12647510 TI - Ephedra and ephedrine for weight loss and athletic performance enhancement: clinical efficacy and side effects. PMID- 12647512 TI - Family size and support of older adults in urban and rural China: current effects and future implications. AB - Change in China's age structure is creating concerns about whether reductions in family size undermine traditional support mechanisms for older adults. Future generations may expect less support as the availability of children declines. In this article, the association between number of children and the receipt of instrumental and financial support is examined for rural and urban populations. Probabilities are modeled as bivariate probits. Coefficients are used to conduct simulations in which support is examined across hypothetical distributions of number of children. The results show that the number of children is an important determinant of support, but future reductions in support may not be as dramatic as anticipated. PMID- 12647511 TI - Appropriate methods for analyzing the effect of method choice on contraceptive discontinuation. AB - The contraceptive method chosen is an important determinant of contraceptive discontinuation. However, method choice is endogenous to contraceptive discontinuation. Using data from the 1997 Indonesia Demographic and Health Survey, we apply a multilevel multi-process model to examine the impact of method choice on three types of contraceptive discontinuation. We confirm that method choice is endogenous to the processes of contraceptive abandonment and method switching, but not failure. Ignoring the endogeneity of contraceptive choice leads to various biases in the magnitude of estimated effects of method choice on abandonment and method switching, but the general conclusions are robust to these biases. PMID- 12647513 TI - Declining sex differences in mortality from lung cancer in high-income nations. AB - After decades of widening, the difference in mortality from lung cancer between men and women has begun to narrow in recent years. Recognizing that the increase in smoking among women relative to men is the proximate cause of the changing sex difference in rates of lung cancer, I analyzed two approaches to identify the more distant sources of the changes. A gender-equality argument suggests that the difference is related to the more general equalization of women's and men's work and family roles, which also encourages the adoption of harmful behaviors such as smoking by women. An alternative explanation suggests that the convergence in mortality from lung cancer among men and women is the byproduct of a lag in the adoption, diffusion, and abatement of smoking by women. Using mortality data on 21 nations from 1955 to 1996, an analysis of logged rates of men's and women's lung cancer mortality and the logged ratio of the rates demonstrated little relationship between the sex difference and gender equality. However, I found a strong and consistent relationship between the sex difference and the stage of diffusion of the use of cigarettes. PMID- 12647514 TI - The joint effects of marriage partners' socioeconomic positions on the risk of divorce. AB - This study investigated the joint effects of spouses' socioeconomic positions on the risk of divorce in Finland. For couples in which both partners were at the lowest educational level, the risk of divorce was lower than could be expected on the basis of the previously documented overall inverse association between each spouse's education and the risk of divorce. Women who were employed or were homemakers, and who had employed husbands, had comparatively stable marriages; couples in which the husband, the wife, or both partners were unemployed had an elevated risk of divorce. A husband's high income decreased the risk of divorce, and a wife's high income increased the risk at all levels of the other spouse's income, but especially when the wife's income exceeded the husband's. PMID- 12647515 TI - Educational attainment and the transition to first marriage among Japanese women. AB - I use data from a large nationally representative survey to examine the relationship between women's educational attainment and the timing of first marriage in Japan. The results indicate that later marriage for highly educated women primarily reflects longer enrollment in school, that university education is increasingly associated with later and less marriage, and that the trend toward later and less marriage is occurring at all levels of educational attainment. These findings are consistent, albeit weakly, with the argument that higher education should be negatively associated with marriage only in countries in which gender relations make it particularly difficult for women to balance work and family. PMID- 12647516 TI - The impact of economic resources on premarital childbearing and subsequent marriage among young American women. AB - This paper extends previous work on premarital childbearing by modeling both the entry rates and the exit rates of unwed motherhood among young American women. In particular, I investigate the impact of economic resources on the likelihood of experiencing a premarital birth and then of subsequent marriage. Using a multiple destination, multiple-spell hazard regression model and a microsimulation analysis, I analyze the accumulating effects of various economic variables. The results show that the economic resources are indeed important both for premarital childbearing and for subsequent marriage. However, the simulations show that large changes in these economic variables do not necessarily translate into large changes in nonmarital childbearing. PMID- 12647517 TI - Cohabiting and marriage during young men's career-development process. AB - Using recently released cohabitation data for the male sample of the National Longitudinal Survey of Youth, first interviewed in 1979, I conducted multinomial discrete-time event-history analyses of how young men's career-development process affects both the formation and the dissolution of cohabiting unions. For a substantial proportion of young men, cohabitation seemed to represent an adaptive strategy during a period of career immaturity, whereas marriage was a far more likely outcome for both stably employed cohabitors and noncohabitors alike. Earnings positively affected the entry into either a cohabiting or marital union but exhibited a strong threshold effect. Once the men were in cohabiting unions, however, earnings had little effect on the odds of marrying. Men with better long-run socioeconomic prospects were far more likely to marry from either the noncohabiting or cohabiting state, and this was particularly true for blacks. PMID- 12647518 TI - Do children from welfare families obtain less education? AB - In this study, we analyzed whether parents' receipt of welfare affects children's educational attainment in early adulthood, independent of its effect through changing family income. We used data from the Panel Study of Income Dynamics with information on parents' welfare receipt over the first 15 years of childhood. Cross-sectional results show that greater exposure to welfare is significantly associated with children's poorer educational attainment. Family fixed-effect regressions also indicate a negative effect of exposure to welfare, but its overall pattern is less consistent. Although exposure to welfare in early childhood has no effect, in adolescence and, to a lesser degree, in middle childhood, its effect is often negative. PMID- 12647520 TI - Parental leave: the impact of recent legislation on parents' leave taking. AB - We use data from the Survey of Income and Program Participation to examine the impact of leave entitlements on unpaid leave usage by men and women after the birth of a child from 1991 to 1999. The results indicate that legislation providing the right to unpaid leave has not affected men's leave usage. The results for women are mixed: in some specifications, leave entitlements are associated with increased leave taking or longer leaves, but the results depend on how we define leave coverage. Our results point to the limited impact of unpaid leave policies and the potential importance of paid-leave policies. PMID- 12647519 TI - Explaining trends in child support: economic, demographic, and policy effects. AB - We use data from the Panel Study of Income Dynamics to examine trends in the receipt of child support (and the determinants of trends) between 1968 and 1997. The findings suggest that political, demographic, and economic forces all exerted downward pressure on child-support payments during this 30-year period, with inflation, the shift to unilateral divorce, and declines in fertility and men's earnings being more important during the earlier years and decreases in men's earnings being more important during the later years. These negative forces were offset by the passage of new child-support legislation in the 1980s and 1990s, including numeric guidelines, universal withholding, and genetic testing. PMID- 12647521 TI - Perspectives. Bush budget: no facts on drug plan, priorities same as in 2002. PMID- 12647522 TI - The changing timetable for the evaluation of new products for formulary consideration. AB - The time for drug manufacturers to educate MCOs about new drugs is not a month before obtaining Food and Drug Administration approval. Today, as Dr. Giaquinta explains, it is imperative that pharmaceutical companies present information about their pipeline products three to five years before drug launch. PMID- 12647523 TI - Employers are cautiously optimistic about health care after election 2002. AB - The Director of Public Policy from the Washington Business Group on Health offers his take on how health care legislation will be addressed in the new Congress. Will the Republican majority work cohesively to reform Medicare? The author believes that "Congress is poised to act on critical legislation to fill some of the gaps in our health care system and has retreated from issues that would only have added more stress to it." PMID- 12647524 TI - Attitudes of physicians toward formularies and services provided by pharmacists. AB - This study was undertaken to evaluate the attitudes of physicians toward formularies as well as services provided by the pharmacy department in a large integrated delivery system (IDS) with a multispecialty group practice. Surveys were sent to all 282 practitioners in the IDS, requesting information on their use of and satisfaction with the existing printed "Formulary Quick List" (FQL), their satisfaction with pharmacy services, and their attitudes toward formularies in general. A five-point rating scale was used. The response rate was 32%, with the majority from staff physicians. Results indicated that practitioners were very satisfied with the services offered by the pharmacists. Their attitudes toward the FQL were very positive. However, their attitudes toward formularies in general were very negative. There was a significant, negative correlation between attitudes toward formularies in general and attitudes toward pharmacy services, as well as toward the FQL. PMID- 12647525 TI - The demise of the small HMO in California. AB - On May 25, 2001, the California Department of Managed Health Care served Maxicare, a Los Angeles-based HMO with approximately 250,000 subscribers, a notice of conservatorship. Within the following 16 months, four other small California HMOs either filed for bankruptcy or were put under state conservatorship. The age of the small HMO in California had come to an end. PMID- 12647526 TI - Can smallpox response teams use the experience of disease management programs? AB - Any attempt to widely disperse smallpox vaccinations will necessitate educating people about the risks and benefits of vaccination. Most disease management programs have extensive experience in distributing educational materials and programs to health care workers and patients as well as in tracking response to interventions. Can this experience lend a hand in the event of widespread vaccination? PMID- 12647527 TI - Preventive care in managed care and fee-for-service plans: is it cost effective? AB - Lack of coverage, knowledge, and physician guidance, and low socioeconomic status are major barriers to receiving preventive care. Compared with fee-for-service insurance, managed care promotes prevention through more generous coverage, physician incentives, and patient education, and members of MCOs use more preventive services than do members of fee-for-service plans. However, cost effectiveness of prevention has not been previously examined. A study of seven preventive services suggests that managed care enrollees more frequently obtained four preventive services, including two less-effective ones. For less-effective services, it may be prudent to base disease-risk assessment on key factors other than age and sex, and screen only patients at high risk. Effective screening strategies must be identified, especially for the underserved, low-income population. PMID- 12647528 TI - [Formation of evolutionary approach as an explanatory principle for ecology]. AB - Although the connection of ecology with evolutionary idea and specifically with Darwinism was proclaimed for a long time it seems that Herbert Spencer's approach with its emphasize on natural equilibrium was much more often used as its real theoretical base. Elements of Darwinian approach appeared only in 1920-30s in works of those few researchers who studying the distribution and population dynamics of different species tried to understand general mechanisms providing their continuing existence. Later, in the middle of 1950s the first attempts were undertaken to consider the population life history (primarily the age specific schedule of death and reproduction) as a result of natural selection aimed to maintain the necessary level of fitness. A special attention in these studies that burgeoned in 1980-90s was paid to looking for various trade-offs between particular parameters of life history, e.g., between the survival of juveniles and fecundity of adults. The problem of life history optimization became central for the whole branch of science named "evolutionary ecology". Though traditionally this branch is connected with Darwinism, it is rooted rather in Spencer's ideas on moving equilibrium and deals more with static than dynamic. Disproportionately less attention was paid to the evolution of communities since these formations could be hardly interpreted as units of Darwinian selection. Moreover, the ecologists dealing with biosphere as a unified biogeochemical system began insist on "nondarwinian" nature of its evolution. The author considers this opinion as not sufficiently grounded. Darwin's ideas about unavoidable exponential growth, intrinsic for any population, consequent deficiency of resources, and differential survival and reproduction of individuals are still useful while studying the evolution of living organisms (phylogenetics) or the development of biosphere as a global ecosystem. PMID- 12647530 TI - [The conflict between optimisation and orderliness in building behaviour of the caddis fly, Chaetopteryx villosa Fabr.(Limnephilidae: Trichoptera), larvae]. AB - The Optimal Foraging Theory predict that an animal should restrict its searching activity to those patches of environment, where the ratio of gain to time and energy spent is maximal. Experimenters usually verify the prediction against the null hypothesis of random activity distribution between patches. As animals always prefer profitable patches to some degree, experimental results can always be interpreted as confirmation of the theory. In opposite to this approach, we put forward the "regularity hypothesis". According to this hypothesis, the finding of a profitable item in some patch makes an animal to stay and test more items within the patch. The readiness to test decreases if the profitability of these items is low, and the animal eventually leaves the patch. It also searches in other patches less carefully. As a result, positive and negative responses to items are repeated by ordered series. In general, this regularity of responses helps animals to choose profitable patches. However, animals may also ignore some profitable patches because of the regularity, so that the behavioural optimazation fails. The regularity hypothesis have been tested in experiment with the search of building material and patch choice by caddisfly larvae. The first of two experimental patches contained egg shell fragments (profitable building material) mixed with a sand (unprofitable). The second patch contained sand only. Larva stayed within the first patch after a shell had been found, so that the probability to find more shells increased. However, larvae started walking after they had found several sand particles. Once starting to walk, they found new shell fragments and tested them, but tended to reject them and eventually leave the patch. Moreover, upon returning to the first patch, larvae also might reject fragments and leave the patch again. Rejections are accounted for by the fact that the duration of testing was too short to identify fragments correctly. As a result, negative responses were repeated to a certain degree independently of the profitability of building material, and the patch choice was not optimal. These results are agreed with the regularity hypothesis. It is argued that the hypothesis can be used as an alternative to the Optimal Foraging Theory. PMID- 12647529 TI - [Immune system and behavioural strategies of reproduction under parasitic pressure]. AB - Since both reproduction and immunocompetence are costly, the negative reciprocal relationships between function were found in many species. Due to reproductive immunosuppression, some fraction of seasonally breeding populations of small mammals reproduces in the first breeding season, while others reproduce in the next one. Enhancement of breeding efforts under the increased risk of mortality and the reproductive delay until recovery are the polar variants of mating behavior of parasitized individuals. However the parasite-induced changes of odor, visual or acoustic signals limit the mating success of the infected hosts. The direct influence of the immune system, regularly activated by infections, to chemicals signals can be answer to the question: why these signals are honest? Decrease in strange infection risk by kin breeding can be a satisfactory strategy of an isolated population. Nevertheless, many species follow the inbreeding avoidance strategy, where the major histocompatibility complex (MHC) genes play key role in kin recognition. The advantage of MHC heterozygosity was found at the all steps of the breeding cycle; including mating choice, fertilization, pre- and postnatal development. So, the relationships between immune system and neuroendocrine regulation of behavior give proximal explanations of the evolutionary stable strategies of breeding behavior. PMID- 12647531 TI - [The problem of individuality in taxonomy]. AB - The ideas of wholeness of superorganism units consist of three constituents: 1) emergeness of properties of the whole; 2) structureness; 3) separability of the whole. Emergeness is a character of both integrated and complex units. Integrated units in strict sense are those that consist of functionally different parts. Species but not super-specific taxa are integrated units. The ideas of unity are based on formula "unity is an aggregation of shape and matter". In taxonomy the idea of archaetype can be used as a shape. Superspecific taxa but not species are recognized as unitary units in taxonomy. Unitary but not integrated units can be considered as individuals. PMID- 12647532 TI - [Classification of nucleotide sequences over their frequency dictionaries reveals a relation between the structure of sequences and taxonomy of their bearers]. AB - Classification of 16S RNA sequences over their frequency dictionaries, both real ones, and transformed ones was studied. Two entities were considered to be close each other from the point of view of their structure, if their frequency dictionaries were close, in Eucledian metric. A transformation procedure of a frequency dictionary has been implemented that reveals the peculiarities of information structure of a nucleotide sequence. A comparative study of two classification developed over the real frequency dictionary vs. that one developed over the transformed frequency dictionary was carried out. The strong correlation is revealed between the classification and the taxonomy of 16S RNA bearer. For the classes isolated, the information valuable words were identified. These words are the main factors of a difference between the classes. The frequency dictionaries containing the words of the length 3 exhibit the best correlation between a class and a genus. A genus, as a rule, is included into the same class, and the exclusion are sporadic. A development of hierarchy classification over the transformed frequency dictionaries separated one or two taxonomy groups, as each stage of classification. The unexpectedly frequent, or contrary, unexpectedly rare occurred of words (of the length 3) in entities under consideration make the structure difference between the classes of the nucleotide sequences. PMID- 12647533 TI - [The geographical parallels of lithorheophil biotic community organisation of small streams of eastern Europe and northern Asia]. AB - The influence of geographical position on taxonomical composition and structure of communities inhabited stony riffles is considered on a material from 79 small streams situated in four regions of Eastern Europe (Moscow region, Southern Ural, Northern Caucasus and Mountain Crimea), as well as on publishing data on Asia and Northern Ural regions. Litoreophil communities of mentioned regions include different but closely allied species with similar habits. Eight basic living forms of litoreophils are common practically everywhere irrespective of geographical position and taxonomical composition of the community. The average number of species in a sample is also constant and includes 13-14 species. Local relief and climate influence the abundance of living forms and species diversity. PMID- 12647534 TI - [Organization and evolution of animal colonies]. AB - Based on the theory of organization and evolution of colonies in an extinct group of hemichordates graptolites (Urbanek, 1960, 1973) the relationship between the events in the late astogeny of bryozoan colonies and their somatic and reproductive cycle. The bryozoan colonies with simple organization are compared with graptoloid colonies and their structure is interpreted within the framework of the morphogen gradient theory. A morphogen produced by the founder-zooid (oozooid) diffuses along the long axis of the colonies and regulates the phenotypic expression of the size and structure of zooids. Evolutionary changes in the graptoloid colonies involve introduction of new characters and their spreading is also accompanied by gradient changes of their manifestation. Evolutionary mechanisms in bryozoan colonies are considered in terms of penetrance and expressivity of genes. In contrast to graptolites, many bryozoan colonies display multiple zones of astogeny changes and repetitions. PMID- 12647535 TI - [Disturbed cooperation of hepatocytes in the rhythm of protein synthesis by chelating agent of cytoplasmic calcium BAPTA-AM]. AB - Previously we demonstrated synchronized oscillations of protein synthesis rate in the hepatocyte cultures upon accumulation of monosialognaglioside GM1 in the medium or after introduction of exogenous GM1 to the medium. The synchronized oscillations of the protein synthesis rate in dense hepatocyte cultures were blocked 30 min after their treatment with 10-20 microM BAPTA-AM--a chelating agent of cytoplasmic calcium. Enzyme immunoassay for GM1 demonstrated similar amounts of GM1 in the medium conditioned for 3 h by dense hepatocyte cultures pretreated with 20 microns]M BAPTA-AM for 1 h and in the medium of normal dense cultures--0.0060 +/- 0.0005 and 0.0055 +/- 0.0005 pmol/1000 cells, respectively. The content of GM1 was also similar in the normal and BAPTA-AM-pretreated hepatocytes--0.158 +/- 0.013 and 0.183 +/- 0.014 pmol/1000 cells, respectively. The synchronized rhythm of protein synthesis has been confirmed in the diluted cultures in the medium conditioned by the normal dense cultures. However, the medium conditioned by the dense cultures pretreated with BAPTA-AM induces no synchronization of the diluted cultures. Since GM1 concentration was normal in this medium, we propose the effect of physicochemical form of the gangliosides accumulated in the medium on their ability to synchronize the rhythm of protein synthesis. PMID- 12647536 TI - Antioxidant properties of heat-treated Hibiscus syriacus. AB - The antioxidant properties of heat-treated Hibiscus syriacus was investigated using DPPH test. The stems and the roots of Hibiscus syriacus were examined, respectively. As a result, the extracts of heat-treated Hibiscus syriacus at 100 degrees C for 24 h were more effective than those of non-treated Hibiscus syriacus in reducing the stable free radical 1,1-Dipheny 1-2-picrylhydrazyl (DPPH). PMID- 12647537 TI - [An organotypic culture of the newt retina together with other tissues of the posterior eye segment as a model for studying the effects of the cell adhesion glycoproteins]. AB - The adult newt retina explanted together with the posterior eye wall and cultivated for a short time in a serum-free medium was tested as an experimental model by several criteria, including the expression of protein markers of the main retinal cell types. Some differences in the expression of specific photoreceptor, interneuron, and glial cell proteins and the localization of acetylcholinesterase activity were found to appear during in vitro cultivation. Using this model, preliminary tests of new cell adhesion glycoproteins from the bovine retina and pigment epithelium were conducted, and the role of pigment epithelial cell proteins in improving cell viability in the cultivated newt retina was revealed. Moreover, the fraction of basic adhesion proteins from the bovine pigment epithelium improved the survival potential of the macroglial (Muller) cell population, compared to that in the control. PMID- 12647539 TI - [Theoretical analysis of inductive effects in higher plant photosynthesis]. AB - The proposed mathematical model of the light stages of photosynthesis in higher plants takes into account the regulatory process of excitation energy redistribution between photosystems depending on the redox state of electron carriers. Nonmonotonic changes in the intensity of pigment fluorescence upon switching on the excitation light (delayed fluorescence induction) are described. The time required for fluorescence to reach its peak intensity and the degree of fluorescence quenching depending on light intensity are analyzed. Th kinetics of light-induced changes in EPR I signal upon switching from the far-red to white light is simulated. PMID- 12647538 TI - [Effect of dietary intoxication by mercury salts on cysteine proteinase activity in rat tissues]. AB - The experiments on dietary intoxication of rats by HgI2 or Hg(NO3)2 show that the activities of lysosomal proteinase cathepsin B and cytosolic Ca(2+)-activated proteinases (calpains I and II) in the liver and kidney depend on the mercury salt solubility and the exposure duration. Mercury iodide and nitrate contribute more to inhibiting cathepsin B and calpains activities in the above tissues, respectively. PMID- 12647540 TI - [The influence of salinity on the rate of dark respiration and structure of the cells of brown algae thallus from the Barents sea littoral]. AB - We studied the changes in the rate of dark respiration (DR) and structure of the cells in Ascophyllum nodosum and Fucus vesiculosus thalluses during the incubation at 40, 34, 20, 10, and 2@1000 salinity for 14 days. The changes in salinity affect the rate of DR and the structure of the thallus apical cells: the organelles swell and later are destroyed. The effect of decreased salinity on the algae was more pronounced as compared to the increase. The stress intensity directly increased with the rate of desalination. Further adaptation of the algae to low salinity enhanced DR and, hence, was an energy-dependent process. Despite higher DR rates (during the stress and adaptation) in F. vesiculosus as compared to A. nodosum, the seaweeds had similar pattern of adaptation to the changed salinity. Different primary response of the seaweeds to 20@1000 salinity was an important exception; apparently, the salinity around 20@1000 is the limit of these species distribution in desalination zones. PMID- 12647541 TI - [Cold hardening of plants is a result of maintenance of an increased photosynthesis/respiration ratio at low temperature]. AB - The capacity of plants for hibernation is ensured by the presence of significant food resources, like in hibernating mammals. In plants, these resources are accumulated in autumn due to maintenance of the rates of true CO2 assimilation elevated with reference to the dark respiration. This becomes possible as a result of higher thermoresistance of photosynthesis, as compared to respiration, and low light requirements for photosynthesis saturation at low positive or negative temperatures. PMID- 12647542 TI - [Architectonic and diagnostic significance of hair cortex and medulla]. AB - Architectonic of hair and spine was studied in 345 mammalian species using scanning electron microscopy. A consistent terminology and system of hair and spine cortex is proposed. The significance of the cortex and medulla structural patterns for taxon identification and adaptation to environmental conditions are discussed. PMID- 12647543 TI - [On the role of morphological and biochemical criteria in species identification: an example of larval dragonflies of the genus Aeshna]. AB - Dragonflies belong to the group of organisms with numerous well-differentiated species-specific characters at the adult stage, on the one hand, and a significantly smaller number or even the absence of such characters at the early ontogenetic stages. An example of the genus Aeshna is used to show difficulties in revealing morphological and biochemical characters allowing identification of larval dragonflies belonging to closely related species of the family. Distinct morphometric characters can be found only in late-instar larvae. The presence of species-specific proteins in the homogenates of thoracic muscles provides the possibility of using biochemical tests for species identification of larvae. Infestation by parasites has no effects on the biochemical parameters studied. Species identification of the early-instar dragonfly larvae is still problematic. PMID- 12647545 TI - [Blood corticosterone concentration and differential leukocyte count in stressed Norway rats with different modes of behavior]. AB - We studied dynamics of leukocyte composition and corticosterone content in the peripheral blood of male Norway rats after long-term selection for tame or aggressive behavior immediately after 20 min restrain as well as 2 and 4 h later. The stressor changed the stressor concentration and the proportion of various types of leukocytes in the both experimental groups. Quantitatively, the changes were more pronounced in the group of aggressive animals. PMID- 12647544 TI - [Interaction with esterases and mechanism of synergistic action of thio- and dithiophosphates containing the fragments of N-acylated glycine and beta alanine]. AB - We studied the interaction between O,O-diethyl-S-[(N-acyl-N alkoxycarbonylalkyl)aminomethyl]thiophosphates and mammalian cholinesterases as well as esterases from insect tissue extracts by kinetic methods and disc electrophoresis. The coefficients of combined effect of these compounds or their dithioanalogs with permethrin were determined. The obtained data suggest that the synergistic effect on the common cockroaches and houseflies is chiefly due to carboxylesterase inhibition by monothioderivatives and monooxygenase suppression by dithioderivatives, respectively. PMID- 12647546 TI - [Territoriality in aquatic animals and their sizes]. AB - The concept of territoriality proposed by E. Odum was used in this study. The territories were determined that were occupied by individuals of specific animal species and the notion "territory of individual" was proposed. The territories of individuals for different representatives of aquatic animals, including fish, were calculated and their relationships with mass and energy potential of animals were established. The territories of individuals and home ranges were compared in invertebrates and vertebrates. It was shown that for the entire animal world the relationship of these areas and mass can be described by a power equation with the exponent indistinguishable from 1. It was established on the example of zooplankton and zoobenthos communities that animal species with a larger size obtain advantage in the oligotrophic water bodies, since when the territories of individuals increase, they are capable of obtaining the necessary amount of food objects at their low density in such conditions. On the contrary, in the eutrophic water bodies, smaller animals have advantage, since they are capable of obtaining the necessary amount of food at a small home range and high density of food objects. A hypothesis was put forward that large and largest aquatic animals can occur in sea and oceanic waters not only because these are huge water basin, but also because these waters are, on the most part of territories, less productive, as compared to continental water bodies. In these conditions, the animals have to hunt over big territories for the necessary amount of food to be obtained and only large animals are able to do this. PMID- 12647547 TI - [On the notion of microrange of soil testaceans (Protozoa: Testacea)]. AB - We studied species composition and population structure of testaceans (Protozoa: Testacea) in a series of soil microblock samples (weighing between 5 and 0.05 g) from podzol horizons L, F, A1A2, and A2 of cowberry-heath-lichen and gramineous cowberry pine forests. Similarity between the testacean communities in the soil microblock samples from horizon A1A2 was at least 0.94 (by Morisita-Horn index of similarity, quantitative data), while in the enzymatic layer of litter it decreased to 0.34. The notion of microrange of soil-forming testaceans is proposed and discussed on the basis of the obtained and published data. PMID- 12647548 TI - [Requirements for glucose, nitrate and phosphate and their variants in the analysis of the mixed culture of Pseudomonas aeruginosa variants]. AB - The calibration data and sensitivity analysis for the variational model of Pseudomonas aeruginosa mixed culture containing R, S, and M phenotypic variants are presented. The model with calibrated parameters (requirements of the variants for glucose, nitrates, and phosphates) adequately describes the experiment. The sensitivity indices (the species abundances and total community size) changed least under the influence of the parameter variations in the one-way limitation areas. The variant proportions are more sensitive to variation of the requirements than the community size. PMID- 12647549 TI - [Multidisciplinary therapy of Tourette syndrome]. AB - The marked fluctuation in symptoms with a spectrum of behavioral problems contribute to misdiagnosis of Tourette syndrome. The authors review the recent progress in diagnosis and management with an emphasis on multidisciplinary approach. Possible associations with various genes have been found in etiology of Tourette syndrome. Development of the disease comes of dopaminerg neurotransmission disorder resulting in cortico-striato-thalamic system dysfunction. Tics are brief movements or sounds that occur intermittently and unpredictably mimicking fragments of normal behavior. Diagnostic criteria are based on the motor and vocal phenomena and their dynamics. The key concept in management are the tic severity scaling correlating with quality of life measurements. Therapeutic interventions indicated at severe alteration in patient's quality of life. Treatment plan combines various drug protocols, psychotherapy and behavioral therapy which should be optimalized for most disabling symptom. Social isolation and self injurious behavior complicates the treatment resistant, severe cases. In these subgroup of patient, an adequate selection of stereotactic intervention could provide an effective control of tic severity or behavioral disorder. Tourette syndrome, as a typical neuropsychiatric disorder, is a striking example for improved efficacy of multidisciplinary approach. PMID- 12647550 TI - [Effectiveness of laparoscopic sacrouterine nerve ablation in our department during the last six years]. AB - INTRODUCTION: Chronic pelvic pain is a very common disease in women. As a final and life-long solution laparoscopic sacrouterine nerve ablation (LUNA) is generally used in the daily practice. OBJECTIVE: To investigate the effectiveness of LUNA by evaluating their results. PATIENTS AND METHODS: Authors performed sacrouterine nerve ablation on 20 patients suffering from chronic pelvic pain. Using visual analogue scale, and with the help of computer database and mailed personal questionnaires efficacy of the method was evaluated. RESULTS: 16 out of 20 cases could be estimated. In 10 patients the level of the pain decreased from 10 to 0, in 2 patients from 8 to 0, while in one from 5 to 0. In 2 patients a decrease from 10 to 4 and 5 was detected in the pain intensity. The level of pain remained unchanged in five patients. There were no side effects during procedures. CONCLUSION: Despite of the low number of the patients authors emphasize the very good effectiveness of laparoscopic sacrouterine nerve ablation in treating of chronic pelvic pain if the procedure is well indicated. PMID- 12647552 TI - [Role of echocardiographic assessment of the left atrial appendage in clinical practice]. AB - BACKGROUND: It is known for a long-while that the left atrial appendage is a frequent source of cardiac thrombus formation associated with systemic embolism. Transesophageal echocardiography made it possible to evaluate accurately the left atrial appendage in vivo. Albeit the most important part of transesophageal echocardiographic evaluation of left atrial appendage is the exclusion of thrombi, growing evidence support that several other clinically important informations can be achieved with its detailed ultrasonographic assessment. AIM: The purpose of authors was to summarize the data available in the literature on the echocardiographic evaluation of left atrial appendage. RESULTS AND CONCLUSIONS: Beyond transesophageal detection of left atrial appendage thrombus it is possible to assess the anatomic variations, dimensions, prethrombotic states and specific flow patterns of the appendage that may have clinically important diagnostic and prognostic impact. PMID- 12647551 TI - [Interstitial boost and altered fractionation as novel approaches to dose escalation in the radiotherapy of glioblastoma multiforme]. AB - INTRODUCTION: The practice of image-based three dimensional treatment planning and conformal radiotherapy techniques give the opportunity to elaborate optimal treatment forms for primary brain tumours. PURPOSE: The authors examined the effect of two novel dose escalation methods on glioblastoma patients. METHODS: In nine cases they treated T1 tumours with single HDR-AL boost of 10-12 Gy dose following the conventional fractionated 60 Gy external beam radiotherapy. In fifteen patients with T2-4 tumours an intensified, hypofractionated regimen with 2.25-2.5 Gy daily and 60 Gy total dose was applied. All the treatments were carried out with image-based conformal methods. RESULTS: Majority of patients endured treatments without neurological deterioration. Transient neurotoxiticy was noticed in one and two cases, respectively. The median survival times (MST) were found to be 17 months (range: 9-25) and 12 months (range: 6-38) in the two groups, respectively. With respect to all patients, the MST was 13 months, while this value in the conventional treatment is generally considered to be 9-10 months. All the three patients who survived more than 18 months was treated with temozolamide chemotherapy as well. CONCLUSION: Based on own experience and current knowledge of authors, it seems reasonable to apply higher biological dose focal radiotherapy and chemotherapy in case of glioblastoma patients with better prognosis. To define the optimal treatment regimens randomised clinical trials should be executed. PMID- 12647553 TI - [Torsades de pointes ventricular tachycardia induced by intravenous amiodarone]. AB - The authors report on the case history of a 61 year old woman with hyperthyroidism induced atrial fibrillation, tachycardiomyopathy and congestive heart failure, in whom life threatening ventricular proarrhythmia (torsades de pointes) developed in response to intravenous amiodarone. The patient in a septic state was resuscitated because of ventricular fibrillation. The atrial fibrillation complicated by a high ventricular frequency was slowed down with intravenous amiodarone; additionally, the iodine-containing antiarrhythmic drug was expected to counter thyrotoxicosis. In response to amiodarone (2 x 300 mg), the sinus rhythm was restored, but the excessive post-cardioversion bradycardia led to the development of extreme QT interval prolongation and torsades de pointes ventricular tachycardias that often degenerated into ventricular fibrillation. In connection with this case, the authors survey those electropharmacological and pathophysiological factors which may have played a role in the emergence of ventricular proarrhythmia based on a lengthening of repolarization through the exhaustion of the repolarization reserve. PMID- 12647555 TI - [Diagnosis of rotavirus infections and their reporting in the Czech Republic in 2001]. AB - In 2001 in 44 laboratories in the CR by laboratory methods 2,250 rotavirus infections were confirmed. From the total number of diagnostic departments 37, i.e. 84.1% use latex agglutination. Among the examined subjects children under 3 years predominated a positive finding being confirmed in 23.0%. The specific morbidity in this age group was 651.3 cases/100,000/year. The hygiene service received in the course of the year 851 notifications of sporadic cases and 72 infections which developed in three epidemics. As to infections confirmed by laboratory examinations only 39.3% were notified. Probably also one imported case of the disease was recorded, no death was reported. PMID- 12647554 TI - [Molecular epidemiology of tuberculosis among prisoners]. AB - The subject of the investigation is a group of 27 strains of Mycobacterium tuberculosis isolated in 2000 from prisoners with the diagnosis of tuberculosis, 19 Czechs and 8 foreigners (mean age 41 and 35 years resp.). The molecular epidemiological examination of these strains was made using the RFLP fingerprint technique (Restriction Fragment Length Polymorphism) with evidence of the insertive sequence IS6110 and the technique of spoligotyping, based on detection of hybridization of spacer oligonucleotides. DNA fingerprinting revealed a high polymorphism in the number and molecular weight of sequence IS6110 which is common in Czech and other European strains of M. tuberculosis. All strains with the exception of two had mutually different fingerprint profiles. In the two with identical fingerprints probably a duplicit examination of the same material was involved which occurred by mistake during transport or in the laboratory. The fingerprint method thus did not prove interhuman transmission of tuberculosis between the examined prisoners. The technique of spoligotyping revealed the finding of genotype Beijing M. tuberculosis in two sick prisoners, one Algerian and one Albanese, and in one Czech prisoner. This genotype found in a high percentage of patients in southeastern Asia and in migrants from this area was detected for the first time in the Czech Republic. The findings are evidence of a satisfactory standard of the programme of tuberculosis control in the Czech prison system and at the same time draw attention to the potential possibility of the spread of tuberculosis from migrants coming from areas with a high prevalence. PMID- 12647556 TI - [Borrelia in ticks biting humans and prevention of Lyme borreliosis]. AB - We examined by dark field microscopy 195 of 209 Ixodes ricinus ticks that were removed from humans in the Czech Republic (mainly in southern Moravia) during 1997-2001. The majority of the ticks were nymphs (62%), 31% were females and 7% larvae; 10.7% of the nymphs, 20.3% of the females but no larvae were found to be infected with B. burgdorferi s.l., the agent of Lyme borreliosis (LB). This observation supports for Central Europe the conclusions drawn from North America about nymphal ixodid ticks as the most important vector stage in transmission of LB. In the course of the year, we found the highest proportion of infected nymphal and female ticks taken off humans in June. The detection of borreliae in a human-biting tick is an important step that enables the general practitioner to prescribe prophylactic antimicrobial treatment and to reduce significantly the risk of Lyme disease in the patient. PMID- 12647557 TI - [Detection of hantavirus antigen in rodents in the Czech Republic]. AB - The work is based on the hitherto implemented examinations of lungs of rodents from the territory of the Czech Republic for the presence of hantavirus antigen. For detection of a species specific antigen the ELISA set Hantagnost from the Poliomyelitis Institute in Moscow was used. A total of 1494 micromammalia of 15 species caught in 1991-2001 were examined. The hantavirus antigen was detected in the lungs of 101 individuals (6.8%) whereby representatives of the species Microtus arvalis predominated markedly: 97 positive from 687 examined (14.1%). Twice the hantavirus antigen was proved in Microtus subterraneus and once each in Apodemus sylvaticus and Clethrionomys glareolus. From hitherto assembled findings ensures that most probably genotype Tula was involved. The authors observed a direct correlation of positivity on the increase of volves and the season of their catching with maxima in the spring and autumn months. No correlation with the sex of the host was found. PMID- 12647558 TI - [Molecular genetics methods in medical mycology]. AB - Molecular genetic methods that use the polymerase chain reaction (PCR) are due their speed widely employed in diagnostic approaches in microbiology. In this report, the possibilities of application of these methods in medical mycology are discussed with regard not only to species identification, but also for genotyping of strains for epidemiological purposes. Recently, a tendency to exploit molecular genetic methods rather for epidemiological studies than for routine species identification may be observed. With regard to the high inter-species variability, careful standardization using samples of isolates of the tested species from corresponding geographical origin is necessary. Perspectives of future development associated with the explanation of molecular biological relations between human tissues and the pathogen, with the recognition of mechanisms of virulence and resistance to antifungal drugs are discussed. PMID- 12647559 TI - [Aspergillus, aspergillosis]. AB - Aspergillus is a ubiquitous fungus which can penetrate into the human body in a natural way, by the respiratory tract or traumatically or iatrogenically. The presence of Aspergillus in the atmosphere is constant, in the environment, especially in moist places or in the neighbourhood of man, in dust. Aspergillus can colonise different substrates, the organic rests, soil, plants, cloting, climatisation, furniture. Aspergillus is in the first phase among the agents responsible for respiratory infections, for different physiopathological mechanisms and for some nosocomial infections. The author gives a review of sources of Aspergillus, especially for hospitalized patients, of mechanisms of the host's defence, of factors enabling the survival of Aspergillus in hosts and of clinical picture. The prevalence of deep forms: aspergillome, allergic bronchopulmonal aspergillosis remain constant. New clinical forms appear due to modern therapy (immunosuppression, organ transplantation). The author describes further laboratory diagnostics with the interpretation of results and epidemiological studies. The early diagnostics of different types of aspergillosis, preventive methods and actual antimycotic therapy can inhibit the progress and improve the prognosis of these infectious diseases. PMID- 12647560 TI - Pragmatic versus form-based accounts of referential contrast: evidence for effects of informativity expectations. AB - Characterizing the relationship between form-based linguistic knowledge and representation of context has long been of importance in the study of on-line language processing. Recent experimental research has shown evidence of very rapid effects of referential context in resolving local indeterminacies on-line. However, there has been no consensus regarding the nature of these context effects. The current paper summarizes recent work covering a range of phenomena for which referential contrast has been shown to influence on-line processing, including prenominal and post-nominal modification, focus operators, and intonational focus. The results of the body of work suggest that referential context effects are not limited to situations in which the linguistic form of the utterance directly specifies the point of contact with context. Rather, context effects of a pragmatic, Gricean nature appear to be possible, suggesting the relationship between linguistic form and context in rapid on-line processing can be of a very indirect nature. PMID- 12647561 TI - Disfluencies signal theee, um, new information. AB - Speakers are often disfluent, for example, saying "theee uh candle" instead of "the candle." Production data show that disfluencies occur more often during references to things that are discourse-new, rather than given. An eyetracking experiment shows that this correlation between disfluency and discourse status affects speech comprehensions. Subjects viewed scenes containing four objects, including two cohort competitors (e.g., camel, candle), and followed spoken instructions to move the objects. The first instruction established one cohort as discourse-given; the other was discourse-new. The second instruction was either fluent or disfluent, and referred to either the given or new cohort. Fluent instructions led to more initial fixations on the given cohort object (replicating Dahan et al., 2002). By contrast, disfluent instructions resulted in more fixations on the new cohort. This shows that discourse-new information can be accessible under some circumstances. More generally, it suggests that disfluency affects core language comprehension processes. PMID- 12647562 TI - Integration of syntactic and semantic information in predictive processing: cross linguistic evidence from German and English. AB - Two visual-world eyetracking experiments were conducted to investigate whether, how, and when syntactic and semantic constraints are integrated and used to predict properties of subsequent input. Experiment 1 contrasted auditory German constructions such as, "The hare-nominative eats ... (the cabbage-acc)" versus "The hare-accusative eats ... (the fox-nom)," presented with a picture containing a hare, fox, cabbage, and distractor. We found that the probabilities of the eye movements to the cabbage and fox before the onset of NP2 were modulated by the case-marking of NP1, indicating that the case-marking (syntactic) information and verbs' semantic constraints are integrated rapidly enough to predict the most plausible NP2 in the scene. Using English versions of the some stimuli in active/passive voice (Experiment 2), we replicated the same effect, but at a slightly earlier position in the sentence. We discuss the discrepancies in the two Germanic languages in terms of the ease of integrating information across, or within, constituents. PMID- 12647563 TI - Studying the grammatical aspects of word recognition: lexical priming, parsing, and syntactic ambiguity resolution. AB - Two experiments are reported examining the relationship between lexical and syntactic processing during language comprehension, combining techniques common to the on-line study of syntactic ambiguity resolution with priming techniques common to the study of lexical processing. By manipulating grammatical properties of lexical primes, we explore how lexically based knowledge is activated and guides combinatory sentence processing. Particularly, we find that nouns (like verbs, see Trueswell & Kim, 1998) can activate detailed lexically specific syntactic information and that these representations guide the resolution of relevant syntactic ambiguities pertaining to verb argument structure. These findings suggest that certain principles of knowledge representation common to theories of lexical knowledge--such as overlapping and distributed representations--also characterize grammatical knowledge. Additionally, observations from an auditory comprehension study suggest similar conclusions about the lexical nature of parsing in spoken language comprehension. They also suggest that thematic role and syntactic preferences are activated during word recognition and that both influence combinatory processing. PMID- 12647565 TI - Revolution and evolution. A brief intellectual history of American psychoanalysis during the past two decades. AB - The past twenty years have witnessed revolutionary changes in the theory and practice of psychoanalysis in the United States. The previously dominant clinical model of ego psychology with its emphasis on ubiquitous unconscious conflict between instinctual drives and the demands of reality and conscience, all mediated by the ego, is now but one of an array of differing theoretical views vying for attention. The alternative competing theoretical and clinical domains can be loosely categorized under the rubrics of object relations, self psychology, and the off-shoots of these which include the relational and intersubjective frames. These insurgent models of mental functioning and development, all of which grow out of object relations theory and thus overlap conceptually, have significant implications for psychoanalytic technique. It is the purpose of this article to review aspects of these models and the historic and revolutionary changes that their appearance have effected in the psychoanalytic landscape. A brief summary will also be made of the more evolutionary advances that have occurred in ego psychology during the past two decades. The extant literature on these issues is complex and voluminous, not to say vast. In advance, I apologize for the necessarily selective and even idiosyncratic nature of this paper. Inevitably, many original and important contributions to the evolving mosaic that makes up contemporary psychoanalysis are ignored or short-changed. PMID- 12647564 TI - Probabilistic constraint satisfaction at the lexical/phonetic interface: evidence for gradient effects of within-category VOT on lexical access. AB - Research in speech perception has been dominated by a search for invariant properties of the signal that correlate with lexical and sublexical categories. We argue that this search for invariance has led researchers to ignore the perceptual consequences of systematic variation within such categories and that sensitivity to this variation may provide an important source of information for integrating information over time in speech perception. Data from a study manipulating VOT continua in words using an eye-movement paradigm indicate that lexical access shows graded sensitivity to within-category variation in VOT and that this sensitivity has a duration sufficient to be useful for information integration. These data support a model in which the perceptual system integrates information from multiple sources and from the surrounding temporal context using probabilistic cue-weighting mechanisms. PMID- 12647566 TI - Mind, medicine, and metaphysics: reflections on the reclamation of the human spirit. AB - Following the publication of such works as Auguste Comte's Cour de Philosophie Positive (1830-1842), in which he argued the inherent immaturity of metaphysical discourse, metaphysics, for Western intellectuals--and especially for Western intellectuals committed to science--has largely been abandoned. In recent years, however, we have seen renewed interest in metaphysics among some researchers and clinicians, due, in part, to increasing attempts to integrate diverse fields of study into some unified and coherent understanding of human life. For many psychologists and psychiatrists, this renewed interest is accompanied by an implicit, and sometimes explicit, re-embrace of the notion of the "human spirit." In this paper we explore some of the processes animating this movement and some of the clinical implications that flow from it. PMID- 12647567 TI - Caring less, doing more: the role of therapeutic detachment with volatile and unmotivated clients. AB - The therapeutic alliance is taught and commonly regarded as essential in engendering change. The formation and maintenance of such an alliance often involves the corrective emotional experience of the patient with the therapist and the therapist's ability to relate empathetically. We posit that some situations and specific clients are better served by therapeutic detachment. In the following case, we put forth the principles and purpose of this therapeutic option. PMID- 12647568 TI - The psychodynamic formulation updated. AB - The goal of this article is to update the conceptualization and format of the traditional psychodynamic formulation to include new areas of knowledge and thereby increase its usefulness. Following a historical review of the concept of psychodynamic formulation, new elements are proposed as essential to update this clinical and educational tool. The traditional formulation format is described along with the modifications necessary to incorporate the new elements. A case example of an updated formulation is offered, including the traditional and new elements, and the potential value and limitations of this new approach are discussed. PMID- 12647569 TI - New mother/old therapist: transference and countertransference challenges in the return to work. AB - Psychodynamic psychotherapists are not generally accustomed to discussing with patients the impact of their own personal experiences on their clinical work. Psychotherapists who return to work postmaternity leave, however, have both a rich opportunity and a clinical mandate to explore the effects of their new parenting on the treatment field. Pregnant therapists have addressed their pregnancy with their patients; however, the experiences of both therapists and patients postmaternity leave are often not discussed to the same extent. Using the author's own experience following her return to clinical work following the birth of her child, this paper explores the ways that the psychodynamic work is affected when the therapist has become a parent, including: changes in transference work; problems of separation and abandonment; and expanding boundaries. PMID- 12647570 TI - Therapeutic boundary issues in working with childhood sexual-abuse survivors. AB - The article describes a study of therapists involved in treating survivors of childhood sexual abuse. The authors focus on what induces participants to change their usual therapeutic boundaries. Through qualitative research utilizing extensive interviews, written questionnaires, and a focus-group discussion, the authors gathered information from therapists related to situations that influence them to change their own boundaries. Emergent themes are discussed and the information is integrated with the authors' experience of supervision and psychotherapy. Concern for client safety is the commonly recurrent issue that pushes therapists to change their boundaries. Other intense feelings, such as resentment of the client, worry about the client's feelings, a wish to connect and imbue hope are also strong influences. Self-disclosure boundaries of the therapist are challenged by the wish to balance power within the relationship. Supervision provides an opportunity to examine what influences therapists to make subtle boundary shifts before they become harmful boundary crossings. PMID- 12647572 TI - An object-relational interpretation of Thomas Mann's Tristan. AB - This paper, which analyzes Thomas Mann's short story Tristan from an object relational perspective, posits that Spinell, the protagonist, is functioning primarily in the schizoid position. Splitting, his primary defense mechanism, is discussed at length. His relationships with others are examined in light of his projection of his internal conflicts, with the idealized, sickly Gavriele representing his artistic, romantic and spiritual side, and her husband, the detested Herr Kloterjahn, representing his more mundane self, Seduced by the romantic notions of Spinell and alienated from her husband, the embodiment of the life principle, Gabriele's health deteriorates, and she becomes a willing participant in her own destruction. Ironic contrasts between Mann's story and the story of Tristan und Isolde, whose narrative informs Mann's story and gives it its title, are explored. It is argued that by the end of the novella, with the death of Gabriele, Spinell is momentarily forced to face reality, but chooses to run from it, and hence is headed for further withdrawal and decline. PMID- 12647571 TI - Behavioral avoidance test for childhood obsessive-compulsive disorder: a home based observation. AB - Obsessive-compulsive disorder (OCD) is one of the most debilitating of the anxiety disorders. As our knowledge about this childhood condition continues to grow, there is a need for controlled treatment-outcome trials with precise assessments that are sensitive to treatment change, to guide the development of effective interventions. To evaluate the efficacy of a treatment protocol, it is necessary to have reliable and sensitive measures of OCD symptoms, including measures of obsessions, compulsions, and related levels of distress and avoidance. Whilst structured diagnostic interviews, semistructured clinical interviews, and self-report measures have been widely used in the assessment of childhood OCD, related levels of behavioral distress and avoidance have not been measured in treatment-outcome trials. This study investigated the sensitivity of a behavioral avoidance test (BAT), conducted in the home environment, in assessing treatment-outcome effects for children and adolescents with OCD following a 14-week cognitive-behavioral therapy (CBT) family intervention, in comparison to children in an 8-week "waitlist" control group. The results of the current study strongly support the sensitivity of a standardized BAT in assessing treatment-related changes in children and adolescents with OCD. Implications and future directions for research are discussed. PMID- 12647573 TI - The relief of a caregiver's burden through guided imagery, role-playing, humor, and paradoxical intervention. AB - Caregiving by relatives to a person with mental illness is demanding and overwhelming. This paper describes how to use guided imagery, role-playing, humor, and paradoxical intervention to help the wife of a man with mental illness in easing her worry, anxiety, and frustration engendered by caregiving. The intervention made the wife feel normal and relaxed in facing the "symptoms" of her husband's mental illness. It also revived the wife's patience, hope, and strength in the process of rehabilitation. PMID- 12647574 TI - To reveal or not to reveal a secret. AB - As the physiological birth of the human infant precedes the psychological birth, the technology that has enabled birth of a child through donor fertility treatments may precede the psychological birth of the parent to such a child. The clinical vignette describes the therapy of a man preoccupied with the secret that his children had been conceived by artificial insemination due to his own infertility. His quandary over whether to keep the secret or reveal it to those close to him, and if so, then when and how, sapped his emotional energy, dragged down his spirits, and unsettled his life. The preoccupation with the secret and the question whether to keep it or reveal it, constitutes an organizing and containing envelope for other concealed issues and unconscious conflicts seeking revelation: who am I and where did I come from? Unsatisfactory answers to these fundamental questions during the pre-oedipal and oedipal stages of development can create a difficult dilemma to overcome. PMID- 12647575 TI - JCAHO: serious nosocomial infections are sentinel events, RCA necessary. PMID- 12647576 TI - Six Sigma boosts quality with ongoing analysis. PMID- 12647577 TI - Think how records might be used before discarding. PMID- 12647578 TI - How's your outpatient 'continuity of care'? PMID- 12647579 TI - Bronchiolitis obliterans: the Achilles heel of lung transplantation. AB - Operative mortality in lung transplantation in many centers has decreased to less than 10%. Despite this improvement in early survival, delayed graft failure related to chronic graft rejection continues to limit the survival and function of lung transplant recipients. Chronic lung transplant rejection Bronchiolitis obliterans (BO) is an inflammatory process that leads to fibrous scarring of the terminal and respiratory bronchioles and subsequent total occlusion of the airways. The histological changes are manifest by a progressive obstructive ventilatory defect on spirometric testing. Histologic diagnosis of BO is difficult due to sampling. The term BO syndrome has been developed to allow clinical diagnosis using spirometric criteria. BO is an alloimmune phenomenon, aggravated by airway ischemia, and infection but predicted by frequent and severe acute vascular rejection. It is characterized by increased expression of TGF beta and other cytokines on airway epithelial cells, increased expression of class II antigens in the airways, lymphocytic bronchiolitis, and bronchial epithelial cell proliferation mediated by numerous cytokines. Airway neutrophilia, and activation of neutrophilis with release of their granules into the airway is a precursor of progressive BO. Loss of normal protective mechanisms--leukocyte antiproteases- against this oxidative insult may be an integral part of disease progression. To date treatment of BOS is infrequently successful. Better understanding of the pathophysiology, and earlier recognition of BO may result in improved long term patient and graft survival and function. PMID- 12647580 TI - [Tooth transplantation: an extra dimension in orthodontics]. AB - In the first part of the presentation, the content of orthodontics has been briefly explained by means of some clinical cases. The main part of the presentation consisted of discussing patient treatment by means of autogenous tooth transplantation with special emphasis on its indications and contra indications. When a tooth transplantation is carried out properly, a success percentage of 90%, even forty years after treatment, can be expected. This high percentage has been reported in a recent study. The success of tooth transplantation is mainly due to the timing of the transplantation and the non traumatic handling during surgery. According to the literature, the ideal moment for tooth transplantation is when half to three quarters of the root has been formed. One of the indications for tooth transplantation is the transplantation of a tooth in the cleft of a cleft palate patient. Two cleft palate patients have been presented in which tooth transplantation was carried out after bone grafting in the alveolar cleft. Finally some findings of a recent study, carried out at our department, dealing with cryopreservation of teeth before transplantation, has been presented. According to the findings of this study, cryopreservation, after removal of the pulp tissue does not inhibit the normal ingrowth of new pulp tissue after transplantation. PMID- 12647581 TI - From X-rays to radiofrequency waves or the changing practice of diagnostic neuroradiology: imaging, health care and economical aspects. AB - Over the past 25 years, radiology has become an increasingly important diagnostic technique in medicine. The majority of radiological techniques still use x-rays, despite the availability of other techniques that do not use ionising radiation. The diagnostic work-up of patients with neurological disorders underwent significant changes in the past 20 years parallel with the advances in medical technology. In neuroradiology, the imaging of the central nervous system, magnetic resonance (MR) imaging has challenged the x-ray procedures such as computed tomography (CT), myelography and angiography. MR imaging uses radiofrequency waves that do not have noxious biological effects. It is generally accepted that MR imaging yields superior image quality compared with CT. Despite the advantages of MR imaging, CT remains an important investigation and has not been replaced by MR. In this memoir the state of the art imaging procedures in diagnostic neuroradiology are reviewed, with their advantages and disadvantages. The failed substitution of CT by MR imaging seems to be mainly due to the limited availability of MR installations and the still long examination times compared with CT. The impact of the changing practice of neuroradiology on health care and the economical aspects are extremely important knowing that financial resources are limited. MR leads to a decrease in invasive diagnostic and therapeutic procedures and the real cost of MR seems to be less than expected. Health care technology assessment and evidence based medicine are less well known in the radiological community, but will become increasingly important in the years to come. PMID- 12647582 TI - [Prevention of twin pregnancy after IVF/ICSI using embryo transfer]. AB - One of the negative aspects of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) is the increased risk of multiple pregnancies. Multiples of high order (triplets and pregnancies of higher order) are obsolete to both doctors and patients and need to be avoided. However, there is an epidemic of twin pregnancies with a higher risk of obstetric, perinatal and neonatal complications than singleton pregnancies and with an important psychosocial, economic and financial impact for the parents to be. A reduction in the number of twins can only be obtained by transfer of one embryo. If this guideline would be applied to all cycles, a dramatic fall in pregnancy rate would occur. In a selected patient group one selected embryo with an excellent implantation potential can be transferred. PMID- 12647583 TI - [ORL pathology resulting from tourism and migration]. PMID- 12647584 TI - [Diagnostic evaluation of "acute" vertigo]. AB - Numerous studies of outpatients have been effected with the object of evaluating the role of the different causes of vertigo, and these have clearly evidenced the importance of the individual disciplinary sectors and, consequently, of the clinicoinstrumental exams necessary in order to reach the correct etiopathogenetic interpretation of the dizzy syndrome. We report the experience of a retrospective study carried out in 310 patients suffering from dizziness who were sent to us from the Emergency Room with a generic diagnosis of "acute vertigo". A detailed medical history was taken in every case, after which the patients underwent a series of otoneurological examinations which enabled us to situate each in the ambit of a vestibular pathology (central and/or peripheral) or to schedule him for a series of specialized exams, the indication of which was drawn directly from the characteristics of the clinical examination and medical history associated with the vertigo symptomatology. Of the 157 cases (50.6%) with otoneurological signs compatible with a vestibular involvement, 77 presented positional paroxysmal vertigo, 34 had an acute peripheral vestibulopathy, and 32 manifested sufferance of the central vestibular structures. In 137 patients, the otoneurological exam was negative and further investigations led to the identification of a psychogenic form in 54 cases, a cardiovascular disease in 31 cases, and a neurological disorder in 24 cases. Our data highlights the fundamental role of the anamnestic approach and the objective examination with particular reference to the search for nystagmic evidence that enabled us to make a definitive diagnosis in an elevated number of cases. In the remainder, we proceeded to perform various instrumental examinations, at times upon the request of the other specialists involved in the management of the vertigo patient. By means of a structured and detailed medical interview, it was possible to obtain useful information that directed our investigation towards a clear distinction between extravestibular and vestibular forms. In this way, furthermore, the same information was able to supply a whole series of clinical indicators of fundamental importance in identifying the most suitable specialist for the clinical picture under examination. A detailed knowledge of the different pathologies (vestibular and extravestibular) capable of causing acute vertigo appears to us to be of fundamental importance if the patients sent from the Emergency Room is to be properly evaluated. From the above, one may see how fundamental a multidisciplinary approach is in appraising the vertigo patients, enabling the correct diagnosis to be reached rapidly and economically, as demonstrated by the results obtained by our group, in which we failed to reach a precise diagnostic conclusion in only 5% of the cases. PMID- 12647585 TI - [Horizontal glottectomy according to Calearo-Teatini for the treatment of tumors involving the anterior commissure of the larynx: oncological and functional results]. AB - Among the different surgical options proposed for the treatment of glottic neoplasms involving the anterior commissure is the Horizontal Glottectomy (HG) proposed in 1978 by Calearo and Teatini. This type of laryngectomy is now included with good reason among the reconstructive horizontal laryngectomies in that it calls for the complete removal of the vocal folds and corresponding thyroid cartilage, with reconstruction of the airways by means of a thyrocricopexy. The object of our study was to describe the indications, surgical technique and oncological and functional results on the basis of the experience acquired by the Authors in 21 patients affected by laryngeal carcinoma (juxta commissural T1a with diffuse hyperkeratosis of the contralateral cord and T1b infiltration to the anterior commissure) who underwent Horizontal Glottectomy (HG) according to the Calearo-Teatinin technique. For 3 of the 21 patients, the follow-up was under 3 years; for 11 patients it was over 5 years; and in 4 cases, it was over 10 years, with a survival rate at 5 years of 91.5%. There were 5 deaths (27.7%), 2 of which were due to non-neoplastic intervening causes, and 1 of which for a recurrence of N (5.5%) a year after surgery. In 3 patients, postoperative RT (radiotherapy) was carried out due to infiltration of the resection borders. With regard to the functional results, the naso-gastric tube was maintained for 8 days on average (min 4, max 12 days), and the tracheal cannula remained in situ an average of 11 days (min 7, max 19 days), while the average hospital stay was 9.3 days. The treatment options currently available for glottic tumors involving the anterior commissure may be divided among radiotherapy (RT), conventional surgery including vertical laryngectomies (Tucker's frontoanterior and Leroux-Robert's frontolateral laryngectomies) and horizontal laryngectomies (Calearo-Teatini's horizontal glottectomy, Majer Piquet's crico-hyoido-epiglottopexy, and Labayle's crico-hyoidopexy), as well as endoscopic laser surgery. Each procedure is adopted on the basis of the experience and philosophy of the surgeon. For tumors involving the anterior commissure, our treatment protocol prescribes HG to be performed whenever possible, as its oncological and functional result have confirmed the validity of the procedure, as long as the indications are followed carefully. Furthermore, when compared to the frontolateral procedure, it provides similar oncological results, but being a much more economical operation on the vertical plane, it avoids stenotic complications, while, compared to the Majer-Piquet procedure, it allows easier and faster rehabilitation, even in elderly patients. PMID- 12647586 TI - [Optic fiber endoscopy in childhood with facial mask]. AB - Flexible fibroscopy is the method of choice in the diagnosis of upper airway (UA) pathologies. Some Authors believe that it should be carried out under local anaesthesia even in children less than one year of age. In this article, the Authors report on their experience with general anaesthesia, using an airway mask that simultaneously allows the flexible endoscope to be inserted and the patient to be ventilated. The study was carried out in the period 2000-2001 in 32 subjects, 18 males and 14 females between 2 and 12 months of age who presented different degrees of stridor and dyspnoea. 18 patients were affected by laryngomalacia, 6 presented bilateral paralysis of the vocal cords, 3 had a paralysis of the left vocal cord, 3 were suffering from congenital anomalies of the epiglottis and 2 had normal larynx from the point of view of morphology and motility. The method described is easy to perform and allows the subglottid area to be carefully evaluated, prolonging, if necessary, procedure time without risking hypoventilation-induced hypoxemia; carrying out the examination on a sedated patient furthermore reduces the likelihood of complications. The Authors therefore recommend that in all infants under one year of age, endoscopy of the upper airways be effected under general anaesthesia with an airway mask, reserving the execution of the same procedure under local anaesthesia to cases involving older children. PMID- 12647590 TI - [Image of the month. Myocardial rupture]. PMID- 12647587 TI - [Case report: ipsilateral association of posterior and anterior canalolithiasis]. AB - Benign paroxysmal positional vertigo (BPPV) is the most frequently occurring dizzy syndrome. The literature provides ample descriptions of the forms determined by canalo/cupulolithiasis of the posterior canal, the most common, as well as canalo/cupulolithiasis of the horizontal canal. Descriptions of the forms of canalo/cupulolithiasis of the anterior canal are, instead, very rare and, sometimes questioned: this is fundamentally due to its superior position, which makes it a much less likely site for the accumulation of debris, both from a conceptual and practical point of view. For these reasons, the Authors feel that the presentation of this case of PPV is of particular interest. After analyzing and excluding other interpretations as less probable, they considered the syndrome as determined by the association of right posterior and anterior canalolithiasis. This form, as in the case of posterior canalolithiasis, responded to treatment with Semont's liberatory procedure. PMID- 12647591 TI - [How I treat...chronic myeloid leukemia]. AB - This review article describes the identification of the tyrosine kinase BCR/ABL as the hallmark of chronic myeloid leukemias (CML) as well as the development of a specific inhibitor of this tyrosine kinase, the STI571 (Glivec, imatinib mesylate). The authors discuss the results of a phase I and three phase II trials reporting the efficacy of STI571 as treatment for CML patients and propose two simplified algorithms that may help to guide decision-making for the individual patient. PMID- 12647592 TI - [Case of the month. Wegener's disease]. AB - A 49 year old woman was admitted because of general poor health condition having lasted for 15 days and been associated for 6 months with sinusitis. During hospitalisation, she developed thrombosis of the retina central artery and signs of cerebral vasculitis. A diagnosis of Wegener disease was suggested based on one side on the presence of multiple excavated pulmonary macronodules and, on the other side, on the signs of cerebral vasculitis. No renal dysfunction was documented in our patient. The detection of serum antineutrophil antibodies (c ANCA and anti-PR3) confirmed the diagnosis of Wegener disease without the need of surgical pulmonary biopsy. In addition to the typical ENT and pulmonary symptoms, this particular clinical presentation of unusual ocular and cerebral symptoms, reminds us that Wegener disease is a vasculitis that can give multisystemic disorders. PMID- 12647593 TI - [Effects of nicotine and cannabinoids on the central nervous system]. AB - We describe the molecular mechanisms of action of nicotine and delta 9 tetrahydrocannabinol, the most potent component of cannabis. Like other drugs of abuse, these compounds enhance dopamine release in a precise area of the limbic system when administered acutely. It has recently been shown that the cannabinoid receptors on which delta 9-tetrahydrocannabinol acts are also activated by endogenous ligands such as anandamide. This unconventional neurotransmitter appears to have important physiological effects in the central nervous system. Both preclinical and clinical evidence suggests that nicotine is more addictive than delta 9-tetrahydrocannabinol. However, the intimate interactions that exist between cannabinoid and opioid systems within the brain suggest that cannabinoids should not be considered as harmless drugs of abuse. PMID- 12647595 TI - [Glycoalkaloids in potatoes]. AB - With its high concentration in starch and vitamins (C, B1 and B2), the potato, cooked in water, is a very interesting source of energy. Unfortunately, it also contains natural toxins: the glycoalkaloids. Their variable amount can depend, in particular, on different environmental factors. This paper summarizes these factors effects and the glycoalcaloids toxicology. PMID- 12647594 TI - [Adverse cutaneous reactions to antidepressants]. AB - Due to the widespread usage of antidepressants by the population, they represent a class of drugs which is at high risk for cutaneous side effects. The diversity in the clinical presentations is large. The variability in their severity is large as well. Morbidity may be quite severe, but the vital prognosis is rarely affected. PMID- 12647597 TI - [Ectodermal dysplasias]. AB - The ectodermal dysplasias form a heterogeneous group of rare and complex genetic diseases with different ectodermal derivates abnormalities. The aim of this paper is to present briefly the anhidrotic form or Christ-Siemens-Touraines's syndrome and the hidrotic form or Clouston's syndrome. PMID- 12647596 TI - [Left ventricular aneurysms: principles of surgical treatment]. AB - After reviewing the historical background, the morphology and physiopathology of left ventricular aneurysms, the authors review the physiology, the technical aspects, and the current surgical indications of aneurysmectomy, with a particular emphasis on the concept of endoventriculoplasty of Jatene & Dor. PMID- 12647598 TI - [How I explore...the secrets of a meta-analysis]. AB - Meta-analysis is becoming a key partner of Evidence-Based Medicine. This statistical tool is currently used in most systematic reviews and allows to quantitatively synthesize the results of several studies by a single weighted average whose precision is indicated by the confidence interval. A standardized graphical representation allows, at first glance, to derive a lot of useful information, especially the number of studies taken into account for the analysis, the mean effect observed in each study with its confidence interval, the relative size (number of subjects) of each study, the homogeneity/heterogeneity of between-study results, finally the overall mean effects with its confidence interval. The meta-analysis plays a key-role in epidemiology, for the evaluation of a risk or a protective factor, and in therapeutics, for summarizing the results of numerous clinical trials with a specific drug or a pharmacological class in a given disease. PMID- 12647599 TI - [Clinical study of the month. Which initial antihypertensive? Results from the ALLHAT trial]. AB - Antihypertensive therapy is well established to reduce hypertension-related morbidity and mortality, but the optimal first-step therapy is still controversial. The "Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial" (ALLHAT) should give such an answer. It is a randomised, double-blind, trial designed to determine whether treatment with either a calcium channel blocker or an angiotensin-converting enzyme inhibitor lowers the incidence of coronary heart disease (CHD) or other cardiovascular disease (CVD) events vs treatment with a diuretic. A total of 33,357 participants aged 55 years or older with mild to moderate hypertension and at least 1 other CHD risk factor were randomly assigned to receive chlorthalidone (12.5 to 25 mg/day; n = 15,255), amlodipine (2.5 to 10 mg/day; n = 9,048) or lisinopril (10 to 40 mg; n = 9,054). The primary outcome combined both fatal CHD and non-fatal myocardial infarction, analyzed by intent-to-treat. Secondary outcomes were all-causes mortality, stroke, combined CHD (primary outcome, coronary revascularization, or angina with hospitalization), and combined CVD (combined CHD, stroke, treated angina without hospitalization, heart failure and peripheral arterial disease). Chlorthalidone was slightly more effective in reducing systolic pressure while amlodipine reduced slightly more effectively diastolic blood pressure. After a mean follow up of 4.9 years, no differences were observed between the three treatments regarding both the primary outcome and the total mortality. Secondary outcomes were similar when comparing amlodipine vs chlorthalidone. A moderately higher 6 year incidence rate of clinically detected heart failure was observed with amlodipine, but without significant influence on mortality. For lisinopril vs chlorthalidone, lisinopril had slightly higher 6-year rates of combined CVD, stroke and heart failure. In conclusion, thiazide-type diuretics are superior in preventing one or more major forms of CVD and offer the advantage to be cheaper. They should be preferred for first-step antihypertensive therapy. However, to reach the recommended blood pressure target, most patients should receive a combination of antihypertensive compounds. Such a combination should always comprise a diuretic agent, in absence of contra-indications. PMID- 12647600 TI - [Clinical study of the month. The ALLHAT-LLT trial]. AB - ALLHAT-LLT was part of the ALLHAT study. The purpose was to determine whether pravastatin compared with usual care reduces all-cause mortality in older, moderately hypercholesterolaemic, hypertensive patients with at least one additional coronary heart disease (CHD) risk factor. 10,355 ambulatory patients, aged 55 years or more, with LDL cholesterol of 120-189 mg/dl (or, 100-129 mg/dl if known CHD) and triglycerides < 350 mg/dl, were randomised to pravastatin (40 mg/d; n = 5,170), or usual care (n = 5,185). Mean age was 66 years; 49% were women; 38% were black and 23% hispanic; 14% had a history of CHD and 35%, type 2 diabetes. Baseline mean total cholesterol was 224 mg/dl; LDL-C, 146 mg/dl; HDL-C, 48 mg/dl, and triglycerides, 152 mg/dl. Mean follow-up was 4.8 years. Among usual care patients, 32% of those with known CHD and 29% of those without CHD started taking lipid-lowering drugs. At year 4, total cholesterol was reduced by 17.2% with pravastatin and by 7.6% with usual care. A random sample had their LDL-C levels assessed: there was a reduction of 28% with pravastatin and of 11% with usual care. All-cause mortality was similar in the two groups (RR, 0.99; 95% CI, 0.89-1.11; p = 0.88), with 6-year mortality rates of 14.9% (pravastatin) and 15.3% (usual care). CHD event-rates were not different between the two groups (RR, 0.91; 95% CI, 0.79-1.04; p = 0.16); 6-year CHD event rates were 9.3% (pravastatin) and 10.4% (usual care), respectively. These results are likely due to the small differential in total cholesterol (9.6%) and LDL-C (16.7%) between pravastatin and usual care, compared with prior statin trials. Such an unusual differential essentially results from the open table of the study and from the possibility of prescribing a statin in the usual care group. PMID- 12647601 TI - [Coping with acute psychological traumas in childhood]. AB - While there is increasing knowledge about predictors of posttraumatic stress reactions in adults, much less is known in children. Importantly, the theoretical integration of research findings and the development of models of post-trauma adaptation have been neglected. After some introducing remarks on the classification and the prevalence of posttraumatic stress reactions, this paper aims to integrate current research findings within a trauma adaptation model that is based on transactional stress theory. Characteristics of the trauma, the child and the social context are assumed to predict posttraumatic adaptation. The effect of these variables on trauma adaptation may be direct or indirect, i.e. mediated by appraisals and coping processes. Based on the proposed theoretical model current research findings are presented and discussed in detail. Finally, the usefulness of the model for the planning and the development of psychosocial interventions after trauma is shown. PMID- 12647602 TI - [Play therapy--psychotherapy with play as the medium: II. New developments]. AB - A wide array of new forms and combinations of play therapy have been developed. The aim of the second part of this paper is to present an overview of these newer approaches, including: focussed therapies for specific disorders; behavioural approaches like the Cognitive-Behavioral Play Therapy and the Parent-Child Interaction Therapy; various combinations with family therapy; and therapies especially for preschool children like Filial Therapy, Developmental Play Therapy and Thera-play. Following a phase of experiments and combinations, the empirical evaluation of many play-therapy forms is needed. Especially questions of the differential indication of specific play-therapies and their effectiveness in the therapeutical practice need to be studied. PMID- 12647603 TI - [Dr. Elisabeth Hecker (1895-1986): career in child and adolescent psychiatry on the one hand--involvement in elimination of the handicapped on the other hand]. AB - During the period of national socialism in Loben/Lublinitz (Silesia) existed a children's special department, where many disabled and mentally retarded children were killed. This was happened under participation of the physician Elisabeth Hecker, who was one of the pioneer's of child and adolescent psychiatry in Germany. During the Second World War she was director of the Loben's clinic for adolescent psychiatry--one of the first of its sort and situated directly in the neighbourhood of the children's special department. Hecker announced children with negative ratings to the "Reich Committee for the Scientific Registration of Severe Genetically and Constitutionally Determined Diseases" in Berlin and transferred them to the children's special department in Loben. In 1979 the German Association of Child and Adolescent Psychiatry appointed Hecker as a honorary member for her commitment after the war. This honorary membership has been discussed critically because of her activities during the period of national socialism. The following paper analyses Hecker's involvement to the child "euthanasia" program based on unnoticed archival sources. PMID- 12647604 TI - [Modification of self-injury and self-mutilation behavior--analysis of a single case intervention study of a boy in a school for mentally retarded]. AB - The present study analysed the self-injurious, destructive behaviour of a mentally retarded boy, ten years of age which was generally noticed through his resistance to demands and because of his self-destructive behaviour and aggressions against others. This behaviour appeared in critical situations (following instructions; not getting something). It is maintained by positive (getting attention) and negative reinforcement (let off demands). The interventions followed the mediating concept, including both female teachers and the mother of the boy. They were instructed to set effective demands, to give positive reinforcement for following rules and to sanction deviant behaviour by over correction. The effects of the intervention were measured through four problem behaviours (oppositional defiant behaviour, self-injurious behaviour, aggressive behaviour against others, evading) with 80 measurements in a multiple baseline analysis (replication over behaviours). The interventions reduced the destructive behaviours of the boy. A follow-up after 10 months proved that no problem behaviours were noticeable anymore. PMID- 12647605 TI - [Environment and health status of the population in areas with high risk of environmental crisis in Italy]. AB - The Italian Ministry of Environment identified 15 areas throughout the country as at "high risk of environmental crisis". The areas host industrial activities that may entail noxious exposures, through occupation and the environment. An epidemiological descriptive study on the residents was carried out using routinely collected mortality data. Mortality data for the period 1981-1994 at the level of municipality were analysed, for around 30 causes of death for both sexes. Methods for small area analysis and disease mapping were used. Rates and SMRs (also adjusted for socio-economic factors) were tabulated for residents and restricted to people born locally. The geographical distribution within areas, time trends, and birth cohort effects were also investigated. Results for some specific areas indicate excesses that can be ascribed to known exposures. The mortality of the residents of areas taken together is substantially higher than expected; significant excesses are more than can be predicted by chance. Due to the heterogenous nature of the risk factors, no common patterns of mortality were observed, but for example 50% of the SMRs for total mortality calculated in the areas, for men and women separately, were significantly greater than unity, and so were 37% of the SMRs for all cancers and 30% for lung cancer. Relative risks were in general of small magnitude, but as populations are large (the areas include some 3.5 million people) and mortality is increased for common causes, absolute numbers of excess deaths were large. These results indicate the occurrence of important risk factors, some of which are related to the industrial activities, but more detailed data are needed for the identification of corrective public health actions. PMID- 12647606 TI - Remission as the goal of treatment in major depressive disorder. AB - The first step in initiating treatment is to establish the diagnosis, confirming MDD and being careful to rule out BPAD and comorbid anxiety or substance abuse/dependence disorders. Consideration should be given to the pathophysiology of the MDD, recognizing that current technology does not allow its identification in an individual patient. The pathophysiology must be considered in the selection of initial treatment; if an initial treatment has failed, treatment with a medication that has a different or expanded mechanism of action should be considered. Clinicians must also consider the importance of psychotherapy, where indicated, and listen to the feedback of their patients, tempering it with a measure of objectivity. Finally, clinicians must constantly query themselves as to whether there has been sufficient improvement in their patients with depression. It is the willingness to be appropriately aggressive in treating MDD that will ensure a timely, optimal outcome for their patients. PMID- 12647607 TI - Remission of depression and the Texas Medication Algorithm Project. PMID- 12647608 TI - Effectiveness measures of major depressive disorder in pharmacoeconomic evaluations. PMID- 12647609 TI - A pharmacoeconomic evaluation of major depressive disorder. PMID- 12647610 TI - The role of pharmacoeconomic modeling in depression management by a health plan. PMID- 12647611 TI - A final checklist. It's never too late--there is still time to take some critical steps to advance your organization toward HIPAA compliance. PMID- 12647612 TI - Managed care's challenges. Health plans use new IT to meet the burgeoning challenges of costs and consumerism. PMID- 12647613 TI - Referral management made easy. Mid-Atlantic IDN assumes responsibility for developing its own automated referral management system. PMID- 12647614 TI - Justifying the RIS before the PACS. A logical migration to a digital and paperless imaging environment requires analysis. PMID- 12647615 TI - What works. Three phases of EDIS. Installation of a comprehensive emergency department information system enables a Virginia IDN to revitalize its ED services. PMID- 12647616 TI - What works. Front-end finesse. A combination of the right technology and adept planning and implementation skills brings financial efficiency to a Phoenix specialty practice. PMID- 12647617 TI - What works. Right on schedule. California healthcare organization finds an automated scheduling system that can keep pace with its increase in patient volume. PMID- 12647618 TI - What works. Good form. A massive hardware replacement generates efficiency for a New York IDN. PMID- 12647619 TI - Multiple users of anytime, anywhere access to information. Virginia IDN extends its use of remote access beyond caregivers to empower its in-house support staff. PMID- 12647620 TI - Bed-der than ever. Pittsburgh hospital uses automated bed tracking and control to speed efficiency in its ED. PMID- 12647621 TI - Managed care hotlist. PMID- 12647623 TI - Long hours can't be defended. PMID- 12647622 TI - Thrive or survive: will new health plan IT needs spur vendor consolidation? PMID- 12647624 TI - Change is good. PMID- 12647625 TI - Mental illness or psychological dysfunction? PMID- 12647626 TI - How can taxes help end Texas' dependence on tobacco? PMID- 12647627 TI - Academic education or licensure to protect the public--where to test? PMID- 12647628 TI - Measuring clinical skills. PMID- 12647629 TI - Eye of the storm. PMID- 12647630 TI - Give a little to get a little? 'Quid pro quo' the key to medical liability caps. PMID- 12647631 TI - Smallpox, big decision. Health care workers must decide whether to be vaccinated. PMID- 12647632 TI - Descriptive study of sexually transmitted diseases in Tarrant County, Texas, from 1998 through 2000. AB - Incidence rates of sexually transmitted diseases in Tarrant County, Texas, from 1998 through 2000 were assessed and compared with the rates in Texas and the United States, with the focus on similarities and differences in gender, age, and race/ethnicity. Data were obtained from the Tarrant County Public Health Department, the Texas Department of Health, and the Centers for Disease Control and Prevention. The rates for gonorrhea and syphilis in Tarrant County were significantly higher than rates in Texas and the United States. The largest disparity was found among blacks, followed by Hispanics and then whites, with those aged 15 to 24 years at greatest risk. To increase awareness and reduce the burden of sexually transmitted diseases, prevention programs need to be developed. PMID- 12647633 TI - Problem-based learning in a managed care seminar for all new residents at an academic medical center. AB - One of the most important challenges in resident education is to train residents how to function in relation to managed care companies and systems so as to enhance the quality, accessibility, and efficiency of health care. Since 1996, The University of Texas Medical Branch in Galveston has presented an annual half day seminar on managed care to new residents. The format involves training sessions and didactic presentations aimed at small groups (led by faculty physicians and nonphysicians from throughout the medical and managed care establishments). Problem-based learning sessions conducted in these groups focus on topics such as the organization of managed care systems, access, network, admit versus observation, inpatient status, denials, avoidable hospital days, concurrent reviews, gatekeepers, referrals, behavioral health, disease management programs, and financial considerations. Pretests and posttests are given to those participating to gauge the effectiveness of the program. In addition, participants complete evaluation forms that can be used by program coordinators to assess resident satisfaction with the learning format and to determine what improvements can be made in the process. For the 1999 and 2000 seminars, posttest results were significantly higher than pretest results for the new residents who participated in the seminar. Each year, seminar evaluations show that the small group format is well received. We conclude that the small-group learning format is effective and enjoyable for the residents and their leaders. The format necessitates the training of 30 group leaders to increase their knowledge of managed care systems. PMID- 12647634 TI - The role of vascular endothelial growth factor (VEGF) and p53 status for angiogenesis in gastric cancer. AB - BACKGROUND: Angiogenesis is of crucial importance for tumor growth and development of metastases. Vascular endothelial growth factor (VEGF) has a potent angiogenic activity and mutations of the p53 gene has been thought to upregulate VEGF. The purpose of our study was to evaluate the prognostic significance of these tumor biomarkers for angiogenesis relative to the information derived from established clinicopathological parameters in gastric cancer. METHODS: In this study, we conducted an immunohistochemical investigation of VEGF and p53 expression in 145 tissue samples obtained from gastric cancer patients undergoing curative surgical treatment. To evaluate angiogenesis, microvessel density (MVD) was counted by staining endothelial cells immunohistochemically using anti-CD34 monoclonal antibody. RESULTS: High MVD was significantly associated with depth of tumor invasion and distant metastasis (p = 0.004, 0.021, respectively). Moreover, overall survival for patients with high MVD were significantly lower than that of low MVD (p = 0.048). Positive expression of VEGF correlated significantly with lymph node and distant metastasis (p = 0.040, 0.048, respectively). However, no significant correlation was found between p53 expression and various clinicopathological parameters. VEGF positive tumors showed a higher MVD than VEGF negative tumors (p = 0.028). The expression of p53 did not correlate with VEGF expression. Also, the relationship between the status of p53 expression and MVD had not statistically significant differences. In the multivariate analysis, status of VEGF, p53 expression and MVD were not an independent prognostic factor. CONCLUSION: VEGF seems to be an important, clinically relevant inducer of angiogenesis and angiogenesis assessed by the MVD may be a useful marker for predicting metastasis in gastric cancer. However, further studies are warranted to clarify the impact of p53 on the angiogenesis and the prognostic significance of angiogenesis in gastric cancer. PMID- 12647635 TI - Characterization of binding and phagocytosis of oxidatively damaged erythrocyte to macrophage. AB - BACKGROUND: Scavenger receptors are thought to be involved in the recognition of oxidized low-density lipoprotein (oxLDL) and oxidized erythrocyte (oxRBC). However, there are controversies about the kind of receptors and ligands related to the binding. Macrophages lacking class A scavenger receptor show identical binding of oxRBC with wild-type ones. METHODS: RBCs were oxidized with ascorbic acid and CuSO4. Lipid oxidation was measured indirectly by measuring TBARS semiquantitatively. The binding and phagocytosis were measured by counting the number of oxRBC bound or taken up after incubation at 4 degrees C or 37 degrees C for 60 minutes to 100 macrophages differentiated from human monocytic leukemia cell line. RESULTS: The degree of oxidation and the binding of oxRBCs were dependent on the concentration of CuSO4. The binding and phagocytosis of oxRBC were inhibited by 99% with oxLDL. Fucoidan, competing class A scavenger receptor, inhibited the binding by more than 90%. The binding of oxRBC was higher at 37 degrees C than at 4 degrees C by 3 times. The binding of oxRBCs was maximal at pH 6.5 and higher than at physiologic pH by 2.8 times. At pH 8.5 and 9.5, binding decreased by 67 and 88%, respectively. CONCLUSION: OxRBCs might bind and be taken up to macrophages not mainly through class A nor B scavenger receptors, but through other scavenger receptors and/or pathways. These processes are dynamic and ionic strength might be involved. PMID- 12647636 TI - Helicobacter pylori and telomerase activity in intestinal metaplasia of the stomach. AB - BACKGROUND: Helicobacter pylori (H. pylori) has been considered a definitive carcinogen in gastric cancer. Telomerase is activated in gastric cancer and some premalignant gastric lesions, including intestinal metaplasia (IM). In this study, we evaluated the relationships of both H. pylori infection and telomerase activity with endoscopic and histologic features in IM. The effects of H. pylori eradication on endoscopic, histologic and biochemical changes were evaluated. METHODS: Endoscopic biopsies were obtained from 43 patients with IM for rapid urease, histologic and telomerase tests. The endoscopic and histologic features, H. pylori infection and telomerase were assessed. After H. pylori eradication, 15 patients were re-evaluated and compared after 4 months. RESULTS: Thirty-four (79.1%) patients were infected with H. pylori. The incidence of H. pylori infection was borderline correlated to the severity of IM (p = 0.076). Telomerase was elevated in eight (18.6%) patients. Telomerase tends to be high in subtype III and endoscopic grade III of IM. After H. pylori eradication, endoscopic extent (p = 0.039) and histologic severity (p = 0.074) showed improvements, and telomerase decreased significantly (p = 0.0001). CONCLUSION: Our data suggest that telomerase is associated with the severity and extent of IM and that H. pylori eradication improves the endoscopic and histologic features in IM, and decreases telomerase activity. H. pylori eradication can be considered one of the methods to prevent gastric cancer in patients with H. pylori-infected IM. Further long-term and large-scaled study will be needed. PMID- 12647637 TI - Adenocarcinoma of unknown primary site. AB - BACKGROUND: Metastatic cancer of unknown primary site occupies 0.5-10% of all diagnosed cancer patients and includes various tumors with diverse responses to systemic chemotherapy. Adenocarcinoma of unknown primary site (ACUPS), the most common subtype, has no standard treatment, rarely responds to conventional treatment and has a poor survival rate. METHODS: The retrospective study was performed to investigate the clinical characteristics and the treatment outcomes of ACUPS. RESULTS: Eighty-one patients with ACUPS diagnosed at Samsung Medical Center from May 1995 to July 1999 were included. The median age was 58 years (range, 29-77). The common sites of metastases were the lymph node, liver, lung and bone in order. In 49 of 81 patients (60.5%), the dominant tumor location was below the diaphragm. The majority of patients (76 of 81) were initially treated with systemic chemotherapy including cisplatin. Responses were evaluable in 70 of 76. Eighteen of 70 patients (25.7%) responded to chemotherapy and complete remission was observed in 6 patients. The overall median survival of 81 patients was 5.6 months. The median survival of the responding patients was 18.3 months but the median survival of the nonresponding patients was 4.6 months (p < 0.01). In univariate and multivariate analysis, age, performance status and response to initial chemotherapy were significant prognostic factors for overall survival. CONCLUSION: We observed poor response to the treatment and survival rate in ACUPS, but complete remission and long-term survival were observed in a small number of patients. PMID- 12647638 TI - A phase II study of vinorelbine, mitomycin C and cisplatin chemotherapy for advanced non-small cell lung cancer. AB - BACKGROUND: This prospective phase II trial was performed to determine the efficacy and toxicity of mitomycin C, vinorelbine and cisplatin combination chemotherapy for patients with previously untreated stage IIIB or IV non-small cell lung cancer (NSCLC). METHODS: Between January 1999 and April 2001, 30 patients with chemotherapy-naive stage IIIB or IV NSCLC were entered into this study. Mitomycin C at a dose of 7 mg/m2, vinorelbine at a dose of 25 mg/m2 and cisplatin at a dose of 75 mg/m2 on day 1 and vinorelbine at a dose of 25 mg/m2 on day 8 were administered. This regimen was repeated every 4 weeks. RESULTS: 29 patients out of 30 patients were assessable. Among the assessable patients, 15 (51.7%) patients had a partial response. The median duration of response and survival was 22 weeks and 39 weeks, respectively. Grade 3 or 4 leukopenia and thrombocytopenia were observed in 28.3% and 4.7% of all the cycles, respectively. Nausea and vomiting of grade 3 occurred only in 2.4% of all the cycles. CONCLUSION: The regimen of mitomycin C, vinorelbine and cisplatin for non-small cell lung cancer is active against advanced NSCLC with tolerable toxicities. PMID- 12647639 TI - Endoscopically observed lower esophageal capillary patterns. AB - BACKGROUND: It has been reported that there are four zones of distinct venous patterns around the gastroesophageal junction (GEJ); i.e. truncal, perforating, palisade (PZ) and gastric zones. Using the distal end of PZ as a marker for GEJ, this study was done to assess the length and patterns of PZ in Koreans, and to assess the prevalence of endoscopic Barrett's esophagus (E-BE) and hiatal hernia (E-HH). METHODS: 847 consecutive patients undergoing diagnostic endoscopy were included. During endoscopy, PZ, squamocolumnar junction (SCJ) and pinchcock action (PCA) were identified. Patterns were classified according to the relationships of the distal end of PZ with SCJ and PCA; A: all three at the same level, B: SCJ proximal to the other two which are at the same level, C: PCA distal to the other two which are at the same level, D: SCJ proximal to the distal end of PZ which is proximal to PCA. Cases with patterns B and D were thought to have E-BE, and those with patterns C and D to have E-HH. RESULTS: Patterns A, B, C and D were 79.2%, 12.1%, 3.8% and 4.9%, respectively. Length of PZ was 3.0 +/- 0.1 cm. E-BE and E-HH were found in 17.0% and 8.7%, respectively. Both E-BE and E-HH were more frequently found in males and in cases with reflux esophagitis. CONCLUSION: E-BE and E-HH are not so infrequent in Koreans as previously thought, if we use the distal end of PZ as an endoscopic marker of GEJ. PMID- 12647640 TI - Longitudinal study of specific antibodies to toluene diisocyanate (TDI)-human serum albumin (HSA) conjugate in patients with TDI-induced asthma. AB - BACKGROUND: An appreciable number of patients with toluene diisocyanate (TDI) induced asthma have high serum levels of specific IgE (sIgE) antibody to TDI human serum albumin conjugate (HSA). A recent investigation suggested a role of specific IgG (sIgG) in the development of TDI asthma. METHODS: We observed the changes in the levels of specific IgE and IgG antibodies to TDIHSA conjugate in TDI-induced asthmatic patients during seven years avoidance. RESULTS: Six subjects with high sIgE and five with high sIgG were enrolled. All of them had taken anti-asthmatic medications with complete avoidance. Serum levels of sIgE and sIgG to TDI-HSA conjugate were detected by ELISA. The level of sIgE continued to decline up to 7 years and the mean half-life was 3.9 years. The mean half-life of sIgG was 4.5 yrs. CONCLUSION: These findings suggest that both sIgE and sIgG to TDI-HSA conjugate may persist for several years after the last exposure to TDI. PMID- 12647641 TI - Pulmonary inflammatory pseudotumor--a report of 28 cases. AB - BACKGROUND: Pulmonary inflammatory pseudotumor is an uncommon benign lesion of the lung. In Korea, most literature of the pulmonary inflammatory pseudotumor was case reports. METHODS: We collected 28 cases of pulmonary inflammatory pseudotumor in Korea. This collective series included 4 cases from our hospital and 24 cases were reviewed from the literature since 1977. The analysis involved the age, sex, chief complaint, hematologic examination, size and location of the lesion, cavity formation, presence of calcification and treatment method. RESULTS: Male was more prevalent (81.5%) than female and mean age was 37.9 years old (6-63 yrs). Chief complaints were cough (44.4%), chest pain (29.6%), fever (22.2%), hemoptysis (15%), sputum (15%) and dyspnea (11.1%). There were asymptomatic cases in 11.1%. Hematologic examination revealed normal finding (53.3%) and anemia (20%). The mean size of the lesion was 4.76 cm (1.5-14 cm) and the locations were parenchymal (85.7%), endobronchial (10.7%) and endotracheal (3.6%). Except the endotracheal case, the lesions were in the right (46.4%), the left (42.8%) and bilateral (7.1%). Calcifications (18.5%) and cavitations (11.1%) were present. Diagnostic methods were open thoracotomy (82.1%), bronchoscopy (3.6%), needle aspiration biopsy (7.1%) and core needle gun biopsy (7.1%). Treatments were surgery (85.2%), steroid therapy (7.4%), rigid bronchoscopic removal (3.7%) and observation (3.7%). Postoperative recurrence occurred in only 1 case (4.3%). CONCLUSION: Pulmonary inflammatory pseudotumor was more prevalent in the male, and patients presented with the respiratory symptoms were common. It was necessary to do surgery in most cases for diagnosis and/or treatment. PMID- 12647642 TI - Gemcitabine therapy in patients with advanced pancreatic cancer. AB - BACKGROUND: Advanced, unresectable pancreatic cancer is an extremely aggressive disease. The 5-year survival rate for pancreatic cancer is only less than 5%. Current therapeutic options for patients with locally advanced or metastatic disease are limited. This analysis is a retrospective evaluation of the efficacy and toxicity of gemcitabine regimen as first-line chemotherapy in patients with advanced pancreatic cancer. METHODS: Seventeen chemotherapy-naive patients with advanced or recurred pancreatic cancer were consecutively treated. Gemcitabine was diluted in normal saline and administered intravenously over 1 hour. Gemcitabine 1,000 mg/m2 was administered once weekly for 3 out of every 4 weeks. RESULTS: The median age of patients was 55 years (range 44-82 years). Based on RECIST criteria, there were 5 cases of stable disease (45%) and 6 cases of progressive disease (55%) among the 11 assessable patients. The median survival time was 189 days (range, 84 to 409 days), the 1 year survival rate was 18% in all 17 patients. Grade 3-4 toxic side effect was leucopenia only (29%) and was easily managed without infection. CONCLUSION: Gemcitabine is well tolerated, but has no objective response in advanced pancreatic cancer. PMID- 12647643 TI - A case of chronic pancreatitis associated with liver infarction and acrodermatitis enteropathica. AB - Liver infarction and acrodermatitis enteropathica are rare complications of chronic pancreatitis. This report shows the case of a 56-year-old man who developed liver infarction due to portal vein thrombosis from chronic pancreatitis and acrodermatitis enteropathica during the course of his treatment. The rare combination of these complications in a patient with chronic pancreatitis has never previously been reported in the literature. PMID- 12647644 TI - A case of pancreatic endometrial cyst. AB - Pancreatic endometrial cyst is an extremely rare instance of ectopic endometriosis that was first described by Marchevsky in 1984. A 21-yr-old woman with a history of epigastric pain and weight loss was found to have a cystic lesion in the pancreas on CT-scan. Under the tentative diagnosis of a pancreatic cystic neoplasm, partial pancreatectomy was performed. Histopathological examination of the specimen revealed cystic endometriosis. The clinicopathological features of the lesion are discussed and literature concerning this extremely rare lesion is reviewed. PMID- 12647646 TI - Papillary muscle rupture after acute myocardial infarction--the importance of transgastric view of TEE. AB - Transesophageal echocardiography was performed to evaluate the exact cause of severe mitral regurgitation in a 64-year-old man presented with hypotension and dyspnea after acute inferior wall myocardial infarction. In mid-esophageal two- and four-chamber view, the ruptured stump of papillary muscle could not be visualized. However, in transgastric two-chamber view, we could clearly visualize the ruptured head of the posteromedial papillary muscle as a separated mass attached by chorda tendinae, as well as the freely mobile stump of the ruptured papillary muscle within the left ventricle. So, the comprehensive transesophageal echocardiography, including transgastric imaging, is always indicated in patients with severe mitral regurgitation after acute myocardial infarction. PMID- 12647647 TI - A case of primary myxoid leiomyosarcoma of the liver. AB - Primary hepatic leiomyosarcoma is very rare and no primary myxoid leiomyosarcoma in the liver has been reported yet. Most cystic space-occupying lesions in the liver are benign in nature. But, rarely, malignancy could appear as a cystic lesion by ultrasonographic examination. A 64-year-old woman with a huge cystic mass detected by hepatic ultrasonography was diagnosed as primary hepatic myxoid leiomyosarcoma by immunohistochemical and ultrastructural studies after various image studies and fine needle aspiration biopsy of the liver mass. PMID- 12647648 TI - Ringing in the new year with the SNAP behavioural risk factors in general practice. PMID- 12647649 TI - Oral contraceptives. PMID- 12647645 TI - A case of refractory Henoch-Schonlein purpura treated with thalidomide. AB - Henoch-Schonlein purpura is an acute, self-limited vasculitis syndrome which shows characteristic skin, joint, renal and gastrointestinal manifestations. It is common in childhood and may also occur in adults with fatal complications such as nephritis and gastrointestinal bleeding. We experienced a case of a 20-year old woman who presented with palpable purpura and severe arthritis. The histopathologic examination of the skin revealed leukocytoclastic vasculitis with perivascular deposition of IgA and she was diagnosed with Henoch-Schonlein purpura. Despite treatment with prednisolone for one month, she had more aggravated purpura and fatal gastrointestinal bleeding. The symptoms were improved shortly by cyclophosphamide pulse therapy with plasmapheresis but symptoms were aggravated and symmetric mononeuropathy of the ulnar nerve developed. She was treated with 400 mg/day of thalidomide and symptoms were improved. We herein report a case of Henoch-Schonlein purpura successfully treated with thalidomide which was refractory to prednisolone, immunosuppressive drugs and plasmapheresis. PMID- 12647650 TI - The birth of the RCGP. PMID- 12647651 TI - Meniere's disease: some patients benefit significantly from SSRIs. PMID- 12647653 TI - Management of fertility issues in cancer survivors. AB - BACKGROUND: With improved survival of cancer patients, the issue of fertility after treatment assumes greater importance. Consideration of fertility preserving measures before treatment needs to be considered. OBJECTIVE: To discuss an approach to management of fertility issues in survivors of cancer. DISCUSSION: Sensitive early counselling on fertility options is important to cancer survivors hoping to achieve a pregnancy. Assessment of fertility, the safety of a possible pregnancy and the impact of cancer treatments on future fertility treatments all need to be considered. PMID- 12647652 TI - Fertility effects of cancer treatment. AB - BACKGROUND: Cancer sufferers are a subfertile group, and most treatments have the potential to adversely affect gonadal function. As cancer treatment becomes more effective and survival rates improve there are more cancer survivors in the reproductive age group for whom parenting is an important consideration. OBJECTIVE: This article outlines the effects on fertility of cancer treatments and techniques to minimise the risk of infertility. DISCUSSION: The overall prospects for younger cancer sufferers to either retain their fertility or have genetic offspring is now better than ever before, due to advances in assisted reproductive technology, the appropriate use of fertility sparing surgery and other techniques to reduce the toxicity of therapy on the reproductive organs. These advances raise new moral and ethical concerns that must be considered before advising cancer sufferers of the options for preserving reproductive capacity. PMID- 12647654 TI - Sexuality and body image after cancer. AB - BACKGROUND: Cancer and its treatments can profoundly affect a person's sexuality including their body image, sexual functioning, relationships, identity and self esteem. One of the more common long lasting effects of cancer treatment is sexual dysfunction, yet for a number of reasons, health practitioners may not adequately address these topics. OBJECTIVE: This article describes the range of factors contributing to psychosexual problems for people with cancer and provides a useful framework for addressing problems and taking a sexual history. DISCUSSION: More than half of all people diagnosed and treated for cancer in Australia now survive their disease. Many of these survivors suffer from significant physical and emotional changes during and after their cancer treatments that affect their sexuality and sexual functioning. Such patients need an opportunity to discuss these changes with their health practitioners, but all too often are not given the opportunity to do so. PMID- 12647655 TI - Second malignant neoplasms following treatment for primary cancer. AB - BACKGROUND: Long term survival following treatment for primary cancer has increased significantly in the past decade. With cure comes 'survivorship'. One of the factors clearly affecting quality and length of survival is morbidity associated with treatment and the development of subsequent malignancies. OBJECTIVE: This article aims to look at the risks associated with the development of second malignant neoplasms, give some meaningful figures, and provide suggestions for follow up. DISCUSSION: The balance of heightened awareness and healthy lifestyle versus relentless surveillance and ongoing patient concern is a difficult one. We hope to make our patients aware of the small but significant risk of a second malignancy, advise on any appropriate screening investigations and encourage a life long relationship with their chosen family physician to enable detection of any future malignancy. PMID- 12647656 TI - Immunisation recommendations following treatment for cancer. PMID- 12647657 TI - Prescribing: what's all the fuss? AB - BACKGROUND: Prescribing is a commonly used skill which has until recently been poorly taught in medical school curricula. This is despite the fact that there are a number of proven approaches to teaching better prescribing. OBJECTIVE: The WHO Guide to Good Prescribing is discussed, with an example elaborating the steps involved in the process. DISCUSSION: Central to this approach is the development of a rational and evidence based list of P- or personal drugs which the prescriber develops familiarity with and uses regularly for specific indications. PMID- 12647659 TI - A GP's duty to follow up test results. AB - Medical negligence claims alleging 'failure to diagnose' are a common cause of claims against general practitioners. In these claims there is often an underlying weakness in the GP's test result and patient tracking systems. This article discusses the duty of care of a GP to follow up patients and their test results. Guidance is provided on how to establish an effective test result tracking system in order to minimise the possibility of a claim arising from 'failure to diagnose'. PMID- 12647660 TI - Health inequalities in general practice. AB - The higher burden of disease associated with low socioeconomic status is an every day challenge faced in general practice. General practitioners can make an important difference to these inequalities in health by being aware of the importance of patients socioeconomic background, identifying groups of disadvantaged patients in their practice population and targeting efforts to improve preventive care coverage and quality chronic disease management to those groups. Divisions are important to this work. The profession also has an important role in advocating on behalf of disadvantaged communities, and joining with other health workers and professions in doing so. PMID- 12647658 TI - Recent advances in the management of chronic heart failure. AB - BACKGROUND: The management of chronic heart failure (CHF) has undergone a revolution in the past decade, highlighted by dramatic turnarounds in opinion, renewed focus on 'traditional' therapies and the emergence of novel diagnostic and treatment options. OBJECTIVE: Based on review of the current evidence, this article aims to provide an update of the issues pertaining to the diagnosis and treatment of CHF. DISCUSSION: Underlying this revolution on the management of CHF have been novel insights into the pathophysiology of CHF. Promising findings from recent clinical trials will also be examined in this article. PMID- 12647662 TI - Test your knowledge. Sudden right eye shadow. Eye series 1. PMID- 12647661 TI - Test your knowledge. Back pain. PMID- 12647664 TI - Gender and the stress response. PMID- 12647663 TI - Bettering the Evaluation and Care of Health (BEACH) 2001-2002. AB - The following overview is designed to disseminate key findings from BEACH, publicize the annual report and to thank the general practitioner participants without whose generous contribution this study would not be possible. PMID- 12647665 TI - GP provision of counselling. More research is necessary. PMID- 12647666 TI - GP visits by health care card holders. A secondary analysis of data from Bettering the Evaluation and Care of Health (BEACH), a national study of general practice activity in Australia. AB - BACKGROUND: Patients of low socioeconomic status are more likely to hold a commonwealth government health care card. Card holders are recipients of age, disability, unemployment or other low income pensions. OBJECTIVE: To compare the general practice managed morbidity of health care card holders with noncard holders. METHOD: Data from one year of the continuous Australian national survey of general practice activity (comprising 98,000 encounters from 980 general practitioners) were used to compare patient and encounter characteristics and the problems managed for health care card holders and noncard holders. Logistic regression adjusted for patient confounders (age and sex, practice size, comorbidity and measured social factors) to describe morbidity associated with health care card status. RESULTS: Health care card holders were more likely to have respiratory, circulatory, musculoskeletal, psychological, neurological, endocrine, digestive, urinary and social problems managed. They were more likely to have chronic and psychosocial problems managed and to receive prescriptions. Female card holders were less likely to have a genital check up (including Pap smear). CONCLUSION: This analysis supports a relationship between socioeconomic status and health. Those from a low socioeconomic status (health care card holders) experience worse psychosocial health and more chronic health problems, have more medications prescribed and receive less preventive care. PMID- 12647667 TI - Audit of a behaviour modification program for weight management. AB - BACKGROUND: The traditional management of overweight and obesity is to make weight loss the primary goal and to offer advice about eating less and exercising more. In controlled settings participants who remain in weight loss programs usually lose 10% of their weight, However, one to two-thirds of the weight lost is regained within one year, and almost all is regained within five years. METHOD: At the Melbourne Weight Management and Eating Behaviour Clinic we designed a program to assist patients achieve and maintain a healthy weight. The program is based on achieving behaviour change rather than focussing on weight loss as the primary goal. The focus is on the reasons why patients exceed their most healthy weight, setting realistic behaviour change and accepting what cannot be changed. DISCUSSION: We conducted an audit of this program to identify changes in eating behaviour and weight management, and examine whether any change were maintained after the completion of the program. PMID- 12647668 TI - Australian doctors and their postgraduate qualifications. AB - BACKGROUND: A key activity for doctors is lifelong learning. This may be as part of professional development, vocational registration, formal postgraduate study, e.g. university diplomas and degrees, or vocational training which leads to fellowship of a professional college. However, there are significant barriers of time and cost that prevent many doctors from participating in lifelong learning. METHOD: We examined the range of basic and postgraduate qualifications held by a representative sample of 1198 doctors provided by the HIC of Australian doctors. DISCUSSION: Significant numbers of general practitioners practise without a formal postgraduate qualification in general practice, unlike our specialist colleagues. However, postgraduate studies are undertaken in a variety of other formats and qualifications. PMID- 12647669 TI - Contamination of interventional research is possible through GP membership of more than one division. AB - INTRODUCTION: General practice divisions were promoted in the 1990s to provide support for general practitioners. Membership patterns are not well understood and may have implications for research and health services development. METHODS: Within a postal questionnaire conducted in 1999, we determined self reported membership of divisions. RESULTS: We obtained a 60% response rate (n = 296) from a random sample drawn from all New South Wales GPs. The majority of GPs (n = 204, 69%) belonged to one division. Thirty respondents (10%) did not belong to any division. Fifty-nine GPs (20%) belonged to two or more divisions, women GPs (n = 27, 31%) significantly more than men (n = 32, 16%) (P = 0.002), and GPs with city or metropolitan area practices (n = 52, 24%) were significantly more likely than rural or remote GPs (n = 7, 9%) (P = 0.005) to belong to two or more divisions. DISCUSSION: If divisions are used as the unit of randomisation for interventional research, there is risk of contamination in study design. Articles reporting such trials should acknowledge this. PMID- 12647671 TI - Times, they are a-changing. AB - The outcome of the US elections in the fall of 2002 serve as symbol of the high degree of change currently affecting the fate of people with HIV. The political and economic environment raises serious concerns about the future of treatment access, support for medical care and the overall priority given the fight against HIV within the federal government. HIV programs, with the exception of research at the National Institutes of Health, have not seen increases in the President's budget since the current administration took office, despite obvious increases in demand. The recent election gives little reason to think that will change. Not only are different attitudes affective every aspect of HIV funding, but different people, with different agendas, have significant influence over federal HIV policy. PMID- 12647670 TI - Prevalence and determinants of HIV-1 subtypes in Canada: enhancing routinely collected information through the Canadian HIV Strain and Drug Resistance Surveillance Program. PMID- 12647672 TI - Anti-HIV drug updates--three drugs on the near horizon. AB - The most significant new information about anti-HIV drugs offered in Barcelona concerned those drugs that are either already available or which will soon be available. This includes new information about T-20 (now called enfuvirtide or Fuzeon) and atazanavir--two drugs which will likely be approved within the next six months. Another new drug likely to be approved soon is FTC (Coviracil), a close relative of 3TC (lamivudine, Epivir), though its importance is less certain than that of enfuvirtide and atazanavir. Important new information was also released about tenofovir (Viread), a drug approved by the FDA late in 2001. Equally important were new observations about some older drugs, particularly the combination of ddI and d4T. Many comparative studies of different drug combinations were also reported, offering new information about the relative value of a number of treatment strategies. PMID- 12647673 TI - New hope from new classes of therapy. AB - A number of very interesting new drugs were discussed in sessions at Barcelona. Two represent new classes of therapy--entry inhibitors and integrase inhibitors- while others seek to offer improvements over drugs in existing classes. Both are welcome advances. PMID- 12647674 TI - Complementary corner. AB - Interest in nutritional health products stems from a number of observations. These include documented nutritional/vitamin deficiencies even in early stages of HIV infection and malnutrition associated with increased risk of HIV disease progression. There is great controversy, however, over whether or not using supplements is always a good idea and if it provides benefits in the long run. There has also been long-standing interest in complementary and alternative medicine (CAM) approaches to managing HIV infection and various conditions associated with HIV. The CAMs most commonly used by people living with HIV are not drugs, herbs or other pharmacologic agents, but rather things like meditation, massage, energy healing, acupuncture and the like. The following article contains summary highlights of studies of nutritional health products and CAM approaches in the setting of HIV presented at the World AIDS Conference in Barcelona. PMID- 12647675 TI - New questions about an old combination--ddI + d4T. AB - For the last several years, the combination of ddI (didanosine, Videx) and d4T (stavudine, Zerit) as a backbone of three-drug therapy has been popular both in treatment and in research. Together, the two nucleoside analog reverse transcriptase inhibitor (NARTI) drugs offered relatively high strength and fairly simple use. Despite this, some researchers have long questioned the wisdom of the combination as it violates one of the key rules of combining drugs: combine only drugs with different side effect profiles. Both drugs are associated with the development of peripheral neuropathy and pancreatitis. Pancreatitis is more commonly seen with ddI and neuropathy with d4T, but both occur to a significant degree with each drug and to a higher degree than was seen with other drugs of their class. However, few if any studies were run comparing the ddI/d4T combination to alternatives such as AZT/3TC (Combivir) or even 3TC/d4T. Both ddI and d4T come from the same company, Bristol Myers Squibb. PMID- 12647677 TI - Interleukin-2 (IL-2, Proleukin) and immune function. AB - IL-2 is an immune-based therapy that results in dramatic increases in CD4+ cell counts when used in conjunction with anti-HIV therapy. Although IL-2 has been discussed in previous issues of PI Perspective, new information warrants a further look at the product. PMID- 12647676 TI - New uses for tenofovir: more questions about d4T. AB - In fall of 2001, the drug tenofovir (Viread) was approved based on data showing its effectiveness in people who had previously developed resistance to one or more of the older anti-HIV drugs of the NRTI class (nucleoside analogue reverse transcriptase inhibitors, such as AZT, ddI, 3TC, d4T, etc.). The drug filled an important niche since tens of thousands of people had been on such drugs for many years already. Since tenofovir also offered the advantage of once daily usage and an excellent side effect record, many people wondered how the drug would fare when used as a person's first therapy or in the early years of treatment. They didn't have long to wait for an answer, which was announced at the Barcelona Conference. PMID- 12647678 TI - Human growth hormone for thymus reconstitution. AB - The thymus is an important immune organ necessary for development of new T-cells (like CD4+ and CD8+ cells.) Without some residual thymus, immune reconstitution with a wide variety of functional CD4+ cells is not believed to be possible. Thus, the state of the thymus in HIV disease and the impact of therapies on the thymus are of great interest to those researching immune restoration approaches. PMID- 12647679 TI - Cell cycles, anti-HIV drugs and treatment. AB - New research regarding how and when anti-HIV drugs are effective (and ineffective) raise questions about many of the combination treatment regimens in use today. The data seem to provide additional insights into why drugs fail, and consequently suggest new strategies for improving the effectiveness of combination therapy. PMID- 12647680 TI - Progress report: organ transplantation in HIV. AB - As people with HIV are living longer due to advances in HIV medicines, there is a rise in death rates from conditions not historically associated with HIV. This includes an increase in risks and rates of both liver and kidney failure, often caused by hepatitis B or C, and underlying kidney disease or HIV-related harm to the kidneys (called HIV-associated nephrotoxicity or HIVAN). Anti-HIV therapies that are processed through the liver or kidney can also, in some cases, worsen these conditions and there have been some instances where the damage to the organ has been wholly caused by the side effects of therapies to treat HIV. For people with very advanced liver disease, liver transplantation is often the only option. People with kidney disease have slightly greater options, including dialysis, which involves being hooked up periodically to a machine to circulate and cleanse the blood. It is critical to assess the effectiveness of organ transplantation in people with HIV in order to determine if it prolongs life, improves quality of life and if so then costs should be covered by third-party payers (insurance, Medicaid/MediCal, etc.). The answers to these questions are not obvious since the kind of surgery associated with organ transplants can be very hard on anyone, let alone people suffering from HIV infection. PMID- 12647681 TI - The challenge of Barcelona. AB - Forty million infected with HIV; five million dead in 2001. Fourteen million children orphaned. And, perhaps most startling, the prediction that seventy million people will have died by 2020 unless there is decisive intervention. The horrifying numbers that describe the international AIDS pandemic provided the backdrop for the 14th International AIDS Conference in Barcelona. It is easy for numbers like these to paralyze those of us who live far from where the epidemic is taking its most deadly toll--in places where HIV treatment, care and prevention programs are more generally available. We wonder what we can possibly do in the face of so much suffering and death. PMID- 12647682 TI - Re-infection: is it a concern for people living with HIV? AB - Re-infection is a term used to describe a new or secondary infection by a virus that has already infected a person. In most viral diseases, re-infection with the same virus doesn't occur because once the immune system conquers the original viral infection, it creates immunity against that virus. Re-infection occurs almost constantly, however, in some types of infection, such as the cold or flu viruses, because each new version of those new viruses is substantially different from the last. This is why a person may develop immunity to the flu strain that is common in one year, but still be at risk from the strain that becomes dominant the next year. PMID- 12647683 TI - We must have presumed consent. AB - More and more people with HIV and/or hepatitis are going to need organ transplants, particularly liver transplants. As more of us all over the world discover we are carrying one or more of these viruses, even if we are being treated for them--or particularly if we are being treated for them--the more likely it becomes that one of our organs is going to cease working effectively. And the longer we are being treated, the longer we live and the more that chance grows. PMID- 12647684 TI - Adolescent sexual trauma screening is urged for HIV-infected women. AB - With one in two women with HIV having a history of being sexually abused before age 18, it's become apparent that this risk factor needs to be addressed by HIV clinics and physicians. Women who have been sexually abused often have multiple social-behavioral problems, including injection drug use and depression, and their ability to adhere to HIV antiretroviral therapy may be dependent on whether their history of abuse is addressed through services and psychological care. PMID- 12647685 TI - Is atherosclerosis connected to PI therapy? AB - In a breakthrough study, researchers at the Columbus Children's Research Institute Center of Developmental Pharmacology and Toxicology have discovered direct toxicity of protease inhibitors to the cells that line blood vessels. PMID- 12647686 TI - CDC prevention project one of few for young MSM. AB - Prevention interventions geared toward young men who have sex with men are few and far between, so clinicians and researchers will be watching with interest next year when some outcomes data are released for one of the most comprehensive prevention projects aimed at this population. A research project of the Centers for Disease Control and Prevention is an intervention that was developed specifically for MSM in the age range of 15 to 25 years. PMID- 12647687 TI - First comprehensive HIV data several years away. AB - Since the Centers for Disease Control and Prevention asked that each state and territory add HIV surveillance to their AIDS surveillance, all but three have complied. California is scheduled to implement HIV surveillance on July 1, Pennsylvania is till going through the process, and Georgia has not announced any plans to implement surveillance. PMID- 12647688 TI - NCI investigators find CD4 t-cell protectors. AB - A senior investigator with the National Cancer Institute discusses in this question-and-answer interview some recent findings that a novel human antibody is effective in preventing some strains of HIV from entering CD4 t-cells. The researchers named the newly identified antibody X5 and found that it reacts strongly against a highly conserved region of the glycoprotein gp120. PMID- 12647689 TI - New viral fitness test helps determine regimens. AB - The enigma of failing antiretroviral regimens and rising viral load in patients who are not progressing to AIDS now has an explanation and a tool that will help clinicians determine which patients will experience this phenomenon. It appears at the same time HIV mutates and becomes resistant to antiretrovirals in some patients, the virus also has a reduced ability to replicate. A new replication capacity test can measure whether the virus has developed a reduced ability to replicate. PMID- 12647690 TI - FDA notifications. Watch out for counterfeit Procrit, 2 lot numbers. PMID- 12647692 TI - FDA notifications. Watch for counterfeit lot of Serostim. PMID- 12647693 TI - FDA warns about counterfeit Epogen. PMID- 12647691 TI - FDA notifications. Product tampering labels Ziagen as Combivir. PMID- 12647694 TI - Computed tomography after breast conserving carcinoma treatment: recognition of surgically induced lesions prior to radiation therapy. AB - To introduce computed tomography for the detection of post-surgical intramammarian lesions, exposing risk factors for subsequent radiation therapy. 104 patients with breast cancer were treated. They previously underwent breast conserving carcinoma treatment and were subsequently irradiated. The techniques were planned individually based on a CTdata set. All the acquired slices were evaluated with regard to seromas, larger tissue defects or organized hematomas. Seromas and tissue defects are raising the rate of byeffects during radiation therapy and deteriorating the cosmetic results. Pretherapeutical CT allows the puncture of hidden seromas and the individual adaptation of the irradiation technique in order to avoid overdosage due to tissue defects. Therefore CT is valuable for all patients prior to radiation therapy. PMID- 12647695 TI - Acute-onset autoimmune hepatitis treated with living donor-liver transplantation. AB - A 50-year-old woman was diagnosed with acute-onset autoimmune hepatitis. She did not respond to steroid therapy including pulse therapy, and was subsequently treated with living donor-liver transplantation 36 days after the beginning of steroid therapy. Except for a period of transient mild acute rejection, her liver function tests remained within a normal range for 2.5 years after the operation. The courses of autoimmune hepatitis patients treated with living-donor liver transplantation have not been previously documented to our knowledge. Living donor-liver transplantation is thought to be one of the therapy options for severe autoimmune hepatitis. PMID- 12647696 TI - Stimulated gastrinemia and maximal acid output in Helicobacter pylori-positive patients. PMID- 12647697 TI - Omentoplasty reinforcement of cervical esophagogastrostomy ansatomoses. PMID- 12647698 TI - [Cytokines. The role of cytokines in colorectal cancer]. PMID- 12647699 TI - Descriptive analyses of the integrity of a US Medicaid claims database. AB - PURPOSE: To examine the integrity of six Medicaid databases for use in pharmacoepidemiology research. METHODS: We performed descriptive analyses to examine four categories of potential data errors: incomplete claims for certain time periods; absence of an accurate indicator of inpatient hospitalizations; missing hospitalizations for those aged 65 years and over; and diagnostic codes in demographic groups in which those conditions should be rare. RESULTS: Prescription claims appeared to be missing intermittently in some states. No valid marker of inpatient hospitalizations could be found for three of six states. Hospitalizations appeared to be missing to varying degrees for those aged 65 years and over. Gross errors in diagnostic codes and demographic data did not appear to be widespread. CONCLUSIONS: Whenever possible, investigators using administrative data should perform macro-level descriptive analyses on the parent data set. In particular, researchers should examine the number of medical and pharmacy claims over time, looking for gaps. Validity of markers of hospitalization should be assessed. The accuracy of diagnosis and demographic data should be examined. Such a descriptive macro-level approach should be used to supplement, and perhaps precede validation of study outcomes using clinical records. PMID- 12647701 TI - Phase II trial of ZD0473 as second-line therapy in mesothelioma. AB - A phase II, open-label, non-comparative, multicentre trial of the platinum analogue ZD0473 as second-line therapy for pleural mesothelioma has been completed. The objectives were to evaluate the activity and tolerability of ZD0473 in patients with relapsed or progressive disease who had received one prior chemotherapy regimen. Forty-seven patients were recruited onto the trial, all aged > 18 years with a life-expectancy > 12 weeks, and World Health Organization (WHO) performance status < or = 2. A starting dose of 120 mg/m2 was administered to 14 patients, six of whom subsequently had their dose escalated to 150 mg/m2. Thirty-three patients received a starting dose of 150 mg/m2. In total, 147 treatment cycles were administered (median number of cycles 3 [range 1-6]). The main toxicity of ZD0473 was haematological (thrombocytopenia) and the most common non-haematological adverse event was nausea. There was no clinically significant nephro-, neuro-, or oto-toxicity. Of the 43 patients evaluable for response, 12% had a minor response (defined by a reduction in lesion size > or = 10% but < 50%), 44% had stable disease, 40% had disease progression, and two patients died before an objective response could be assigned. Median time to progression and death in evaluable patients was 77 days (95% confidence interval [CI]: 44, 105 days) and 203 days (95% CI: 165, 277 days), respectively. In conclusion, although ZD0473 demonstrated a manageable tolerability profile, no complete or partial responses were seen in second-line treatment of mesothelioma. This trial also demonstrates that clinical trials in second-line mesothelioma patients are feasible. PMID- 12647700 TI - Patient-centered diabetes self-management education. AB - Diabetes self-management education (DSME) has been shown to improve health outcomes. Yet, relatively little is known about how DSME has its effects. Literature reviewed from the past 3 years indicates that if DSME is to become more effective interventions need to be theory-based, to increase patient involvement in their care, and to encompass a broader array of evidenced-based outcomes. Outcomes reviewed go beyond knowledge and glycemic control to include prevention of diabetes, quality of life, and reduction of cardiovascular risk. The ability of practitioners and health care systems to implement, adopt, and maintain patient-centered interventions over time is discussed. By linking theory to behavior, and broadening the outcomes examined, advances can continue to be made in closing the gap between the scientific base for the treatment of diabetes, and the care and outcomes patients experience. Further research on patient-centered approaches that promote self-management is seen as critical in closing this gap. PMID- 12647702 TI - [Deeply carry out studies on diagnosis and treatment of traumatic optic neuropathy]. PMID- 12647703 TI - Calcium Summit II. Washington, DC January 17, 2002. PMID- 12647704 TI - Pre-eclampsia, mechanical heart valve prosthesis and prematurity. PMID- 12647705 TI - Irreducible uterine torsion at caesarean section: how to deliver? PMID- 12647706 TI - Silent dehiscence of a caesarean section scar with placenta praevia accreta. PMID- 12647707 TI - Acrania. PMID- 12647708 TI - Maternal group B streptococcal meningitis in the postpartum period. PMID- 12647709 TI - Intrapartum subcutaneous emphysema and pneumomediastinum. PMID- 12647710 TI - Hypersensitivity to Mirena--a rare complication. PMID- 12647711 TI - In conisation with pelvic lymphadenectomy adequate for FIGO stage 1B1 cervical cancer? Case report and review of the literature. PMID- 12647712 TI - Viable retained fetal bones and secondary infertility: ultrasound and hysteroscopic appearances. PMID- 12647713 TI - Cervical endometriosis presented as a polypoid mass of portio cervix uteri. PMID- 12647714 TI - Scar endometrioma: a cause for concern in the light of the rising caesarean section rate. PMID- 12647715 TI - Recurrent abortion caused by Haemophilus influenzae with subsequent successful pregnancy. PMID- 12647716 TI - Uterine diverticulum as a late complication of caesarean section. PMID- 12647717 TI - Bladder leiomyoma--a rare cause of bladder symptoms. PMID- 12647718 TI - Management of an extremely large cystocoele. PMID- 12647719 TI - Sarcoma of the uterus metastasising to the spleen--an unusual cause for splenomegaly. PMID- 12647720 TI - Ovarian carcinoma detected by routine cervical smear. PMID- 12647721 TI - Clarification added to hospital standards addressing contracted patient care services. PMID- 12647722 TI - "Disaster" replaces "emergency" in "medical staff" privileges standard. PMID- 12647723 TI - Fees apply to all sentinel event review alternatives. PMID- 12647724 TI - Credentialing revisions reflect reduced expectations for networks. PMID- 12647725 TI - 2002 Directory of air medical programs. As of August 19, 2002. PMID- 12647727 TI - Current awareness in geriatric psychiatry. PMID- 12647726 TI - The nurturers. Interview by Jay Greene. PMID- 12647728 TI - Advanced beneficiary notice [ABN] requirement. PMID- 12647729 TI - Surgical treatment of esophageal cancer. PMID- 12647730 TI - Mass treatment of filariasis in New Guinea. PMID- 12647732 TI - High-frequency ventilation. PMID- 12647731 TI - [Congress culture today--an analysis of scientific contributions at the 2002 German Orthopedics Congress]. PMID- 12647733 TI - [Acupuncture in chronic back pain. Background, development and design of the German Acupuncture Trial (gerac-cLBP)]. PMID- 12647735 TI - Typhoid fever. PMID- 12647734 TI - [Acute backache guideline clearing process. Methodology and results of formal evaluation]. PMID- 12647736 TI - Typhoid fever. PMID- 12647737 TI - [Differences in metabolism of chondrocytes of the knee and ankle joint]. PMID- 12647739 TI - Formation of the International Federation of Professional Aromatherapists (IFPA). An interview with Linda Orton-Jay. PMID- 12647738 TI - [German patients as experimental rabbits. Spiegel Magazine reports on implantation of hip endoprostheses with ROBODOC]. PMID- 12647741 TI - The European Society of Cardiology XXIV Annual Congress 2002. Berlin, Germany, 31 August-4 September, 2002. PMID- 12647742 TI - American Heart Association Scientific Sessions 2002. Chicago, IL, USA, November 9 12, 2002. PMID- 12647740 TI - Introduction: a welcome to the First Special Animal Health Issue of AAPS PharmSci. AB - The goal of this special volume is to provide veterinary scientists with state-of the art reviews in animal health and to inform human health scientists of the various challenges and collaborative opportunities associated with their animal health counterparts. The contributors are highly respected experts, providing invaluable insights into current issues and state-of-the-art advances within veterinary medicine. PMID- 12647743 TI - Introduction: pharmaceutics and drug delivery. PMID- 12647744 TI - Current bibliography of the history of science and its cultural influences. 2002. PMID- 12647745 TI - Software and technology: critical tools for implementing evidence-based treatment. PMID- 12647746 TI - Health care fraud and abuse: market change, social norms, and the trust "reposed on the workmen". AB - Health care fraud and abuse reportedly account for 10 percent of total spending on health care, or about $120 billion per year. Not surprisingly, Congress has granted fraud control personnel sweeping powers with which to attack the problem. Unfortunately, effectively addressing health care fraud is exceedingly complicated, particularly in light of recent major changes in the medical marketplace and the social context of such conduct. Broadly speaking, physicians view such conduct as essential to ensure high-quality care; program administrators view it as the price of the program; fraud control personnel view it as criminal misconduct; and the public's view depends greatly on who is benefitting. Social norms regarding health care fraud vary among these groups as well. The article examines the practical and theoretical challenges associated with attacking health care fraud and the merits of the current fraud control regime in light of these considerations. PMID- 12647748 TI - Managed care, assisted suicide, and vulnerable populations. PMID- 12647747 TI - Hospitalists and the doctor-patient relationship. AB - Hospitalists--physicians whose practice focuses on the care of hospitalized general medicine patients--are increasingly common in the United States, often displacing primary care physicians from this role. While advocates of hospitalists point to evidence of cost reduction and perhaps improved short-run outcomes, critics question whether costs or long-run outcomes are improved and whether there may be insidious effects on the doctor-patient relationship. I define a framework for addressing these questions, assess the available evidence, and identify a research agenda to better understand the implications of the hospitalist movement for the doctor-patient relationship. Using a framework that emphasizes general and patient-specific knowledge as crucial to a successful doctor-patient relationship, I argue that the success of the hospitalist model will depend on its ability to continue to develop its areas of technical expertise while ensuring care that is both continuous and appropriate to the needs and values of individual patients. PMID- 12647749 TI - Law as a structural factor in the spread of communicable disease. PMID- 12647750 TI - AIDS as a weapon: criminal prosecution of HIV exposure. PMID- 12647751 TI - The Texas Communicable Disease Prevention and Control Act: are we offering enough protection to those who need it most? PMID- 12647752 TI - Epilepsy and pregnancy. PMID- 12647753 TI - Policy analysis at the Department of Health and Human Services, then and now. PMID- 12647754 TI - FDA halts 27 gene therapy trials after illness: leukemia-like cases in 2 children in France prompt the action. PMID- 12647755 TI - 30 years after abortion ruling, new trends but the old debate. PMID- 12647756 TI - How one clone leads to another. PMID- 12647757 TI - Drug sales bring huge profits, and scrutiny, to cancer doctors: insurers and experts see high costs and conflicts. PMID- 12647758 TI - Health data monitored for bioterror warning. PMID- 12647759 TI - The neurologist and stroke. PMID- 12647760 TI - Proceedings of the International Conference on the Role of Health Professionals in Addressing Violence Against Women. 15 October 2000, Naples, Italy. PMID- 12647761 TI - A sudden bleeding meningioma. PMID- 12647763 TI - Seeking health care information: most consumers still on the sidelines. AB - Contrary to popular belief that Americans avidly seek health information- especially on the Internet--a majority of Americans in 2001 sought no information about a health concern, according to a Center for Studying Health Systems Change (HSC) study. And, instead of surfing the Internet, the 38 percent of Americans who did obtain health information relied more often on traditional sources such as books or magazines. People living with chronic conditions were more likely to seek information, yet more than half did not. Education is key to explaining differences among people. Those with a college degree are twice as likely to seek health information as people without a high school diploma. As consumers are confronted with more responsibility for making trade-offs among the cost, quality and accessibility of care, credible and understandable information will be critical to empowering consumers to take active roles in managing their care. PMID- 12647762 TI - Delayed angiography alone is not enough to discard arteriovenous malformation after haemorrhagic stroke. PMID- 12647764 TI - European urinalysis guidelines. PMID- 12647765 TI - Interpreting reaction time measures in between-group comparisons. AB - Although reaction time measures have been used extensively in many types of between-group comparisons, the assumptions and limitations of reaction time measurement are not always recognized. In this article we discuss three issues that should be considered when designing and interpreting comparisons involving reaction time. These concern speed-accuracy tradeoffs, methods of analyzing measures postulated to reflect specific processes, and methods for distinguishing group-related influences that are shared with other variables from those that are unique to a single variable. PMID- 12647766 TI - Cognitive experimental approaches to investigating impaired cognition in schizophrenia: a paradigm shift. AB - There has been a growing interest in cognitive deficits in schizophrenia in recent years. The use of traditional clinical neuropsychological measures of cognition in schizophrenia has the advantages of normative data and standardized administration, and remains the standard for cognitive assessment in clinical practice. However for the investigation of cognition in schizophrenia these methods have critical limitations in their ability to measure specific cognitive processes. In this paper we discuss solutions to these interpretive and measurement issues that can be provided by measures derived from experimental cognitive psychology. We suggest that the use of these measures will increasingly become the preferred approach to the investigation of the nature, neurobiology and genetics of impaired cognition in schizophrenia in the coming years. PMID- 12647767 TI - RT and non-RT methodology for semantic priming research with Alzheimer's disease patients: a critical review. AB - This paper provides a critical review of RT and non-RT methodology for the conduct of research on semantic priming in Alzheimer's disease (AD) compared to elderly normal (EN) individuals. The review is organized by type of semantic priming (or attribute-knowledge) paradigm. Theoretical as well as methodological issues are highlighted insofar as they are relevant to: (1) the design, conduct, and analysis of semantic priming experiments (and closely related types of experiments) with AD compared to EN participants, and (2) the interpretation of findings from these experiments. Recommendations are provided for further research involving semantic priming and related paradigms with AD and certain other neuropsychological populations. PMID- 12647768 TI - Reaction time performance in adults with HIV/AIDS. AB - Infection with the Human Immunodeficiency Virus-Type 1 (HIV-1) has been shown to lead to cognitive decline in a substantial proportion of infected adults. The characteristic neuropsychological symptomatology includes memory dysfunction, higher order attentional disturbance, executive dysfunction and, most relevant to this manuscript, cognitive slowing. This paper reviews the extant literature on reaction time (RT) performance in HIV-infected adults with an emphasis on mental chronometry. In addition to discussing studies of simple and choice RT, we also examine the utility of RT paradigms in the assessment of selective attention, covert orienting, sustained attention, divided attention, working memory, and implicit memory. Studies documenting the utility of RT tasks to track treatment response are also introduced. In aggregate, research to date that has employed RT tasks in the evaluation of HIV-infected patients has found that HIV infection leads to a mild degree of cognitive slowing that tends to worsen with increasing disease severity. It needs to be noted, however, that a significant percentage of studies failed to find HIV infection to lead to RT slowing. Results of Brinley plot analyses, a technique that shares commonalities with meta-analysis, reveal that HIV-infected patients diagnosed with AIDS are on an average 22% slower than uninfected controls. This paper also reviews data that has shown that HIV associated cognitive slowing, as indexed by single and dual choice RT, is amenable to pharmacologic intervention using the psychostimulant methylphenidate. Given the demonstrated sensitivity of RT tasks to the neuropsychological effects of HIV infection, it is recommended that RT tasks be included whenever possible in the routine clinical evaluation of HIV-infected patients. PMID- 12647769 TI - Locating the human frontal eye fields with transcranial magnetic stimulation. AB - The variability in the location and function of the human frontal eye fields (FEFs) was assessed using transcranial magnetic stimulation (TMS). Ten subjects performed a saccadic eye movement task previously shown to be influenced by TMS of the FEFs. A sequence of points over the prefrontal cortex was stimulated until an effective site of the TMS was found that induced contralateral saccade delays. In 7 out of the 10 subjects, we were able to localize a region in the prefrontal cortex that when stimulated produced delays in the execution of contralateral saccadic eye movements. The location of this functionally defined FEF region across these subjects was approximately 1.5 cm anterior to the motor hand area, although there was considerable variability in this measure. In the remaining 3 subjects, no site within our circumscribed probing was found that when disrupted with TMS produced delays in contralateral saccadic eye movements. The inter individual differences in the location and function of the FEFs highlights the importance of using functional as well as anatomical landmarks when attempting to localize brain structures. PMID- 12647770 TI - Using reaction time to assess patients with unilateral neglect and extinction. AB - Brain damage often results in visual defects and/or higher order visuo-spatial disorders including the syndromes of unilateral neglect and extinction. These syndromes and their associated behavioral sequelae are described along with several standard assessments and methods for behaviorally quantifying such deficits. In particular, the advantages of the reaction time measures commonly used by cognitive neuroscientists are surveyed. In order to illustrate how reaction time measures can be used to examine the subtleties of visuo-spatial deficits, several reaction time studies that have been conducted in patients with neglect and/or extinction are discussed. These studies stand as an example of how reaction time measures can contribute to both clinical and experimental neuropsychology. PMID- 12647771 TI - Orienting attention in aging and Parkinson's disease: distinguishing modes of control. AB - Past research on attentional orienting and Parkinson's disease (PD) has been compromised because the experimental paradigms tended to confound different forms of orienting. We sought to overcome this by examining the attentional orienting of three distinct groups (PD-patients, age-matched controls, and young controls) on five different tasks, four of which isolated pure forms of orienting. On two covert orienting tasks PD patients oriented volitional (Experiment 1) and reflexive (Experiment 2) covert attention in a healthy and normal manner for their age. On two overt orienting tasks, PD patients were found to execute volitional eye movements that were prone to undershoot their target goal (Experiment 3), and reflexive eye movements that were unusually fast (Experiment 4). When required to perform an antisaccade task (Experiment 5), which combines reflexive and volitional modes of overt orienting, PD patients performed normally. This indicates that using a task which combines different modes of orienting creates a situation that is more than the sum of its parts. Together our study supports the thesis that it is crucial to isolate and investigate different modes of attentional control. PMID- 12647772 TI - Mental chronometry in the study of individual and group differences. AB - The papers in this issue are excellent examples of the many uses of measuring reaction time in the exploration of nervous system pathologies. In our commentary we consider mental chronometry as a field that seeks to measure the time course of mental operations in the human nervous system. We draw upon diverse methods such as neuroimaging, electrical recording and reaction time to illustrate the use of chronometry in conjunction with anatomy and genetics to approach both normal individual differences and pathologies. The goal is to examine general and specific changes in neural networks that underlie both variations in normal function and changes due to pathology. Although much remains to be done along these lines, it is now possible to see how the various chronometric contributions outlined in this special issue can converge to provide a basis for improved understanding of the genetic and experiential basis of cognitive and emotional processes. PMID- 12647773 TI - Evaluation of acrosome reactivity using the Acrobeads test in varicocele patients: findings before and after treatment. AB - The Acrobeads test was performed on semen samples from 43 patients with varicocele before and after varicocelectomy. Sperm motility significantly increased after surgery (p <.05), while sperm concentration and motile sperm concentration did not alter postoperatively. Sperm motion analysis using CellSoft 3000 did not demonstrate a significant change after treatment. Acrobeads score significantly increased after the procedure (p<.005). Pregnancy was achieved in 10 patients (28%). Acrobeads score in cases that achieved pregnancy was increased postoperatively (p<.005). The percentage of patients with a postoperative increase in Acrobeads score in the group that achieved pregnancy was significantly higher than that observed in the unsuccessful group (p <.05). Sperm parameters other than the Acrobeads score did not show a significant difference between the successful and unsuccessful patients. The Acrobeads test assessed postoperatively can be useful in precisely evaluating fertility potential after varicocele repair. PMID- 12647775 TI - Molecular biology of male infertility. AB - About 15% of couples have reduced fertility and in approximately one-half of all cases the reason is male infertility, usually of genetic origin. Thus, in the context of research in genes involved in reproduction and sex determination, genetic anomalies in gametogenesis are being extensively studied. The most frequent pathogenic causes of male infertility are Y-chromosomal microdeletions (8-15%) in the long arm of the Y chromosome, which, by loss of specific DNA segments, leads to loss of vital genes for sperm production. Infertile men, who attend infertility clinics, rise to 15% among those with azoospermia or spermatogenesis problem. The new technique of intracytoplasmic sperm injection has allowed many infertile men to achieve their dreams of fatherhood. However, the spermatogenic defect is genetic anomalies, which might be a potential risk of transmitting this defect to future offspring. Therefore, genetic counseling of all couples with the diagnosis of male infertility is recommended before their enrolment in intrauterine insemination, in vitro fertilization, and intracytoplasmic sperm injection. The important role of genetic abnormalities in the causation of human male infertility is increasingly recognized. While much remains to be learned in this fast-moving field, considerable progress has been made in the clinical delineation of genetic forms of male infertility and in the characterization of the responsible genes and their mutations or deletions. This review should provide insight into the understanding of parthenogenesis of male infertility in the human. PMID- 12647774 TI - Erectile dysfunction: current concepts and future directions. AB - Major advances in science and medicine have led to improved understanding of the pathophysiology of erectile dysfunction. The development of reliable pharmacological therapy for erectile dysfunction has led to heightened awareness in the public and medical communities. This article reviews recent clinical advances and future research directions. PMID- 12647776 TI - Inhibitory mechanisms of lead on steroidogenesis in MA-10 mouse Leydig tumor cells. AB - Lead can directly influence Leydig cell steroidogenesis, which results in reduction of testosterone and causes low sperm counts in human beings and animals. This study investigated the effect of 6 h incubation time of lead on steroidogenesis in MA-10 mouse Leydig tumor cells. Lead acetate, ranging from 10( 8) to 10(-5) M, caused profounder inhibitory effects on human chorionic gonadotropin (hCG)- and dibutyryl cAMP (dbcAMP)-stimulated progesterone production for 6 h in MA-10 mouse Leydig tumor cells. Lead acetate significantly inhibited hCG- and dbcAMP-stimulated progesterone production from 20 to 35% in MA 10 cells at 6 h. Lead suppressed the expression of steroidogenesis acute regulatory (StAR) protein from 30 to 55%. Moreover, the activities P450 side chain cleavage (P450scc) enzyme and 3beta-hydroxysteroid dehydrogenase (3beta HSD) were reduced by lead from 15 to 25%. Thus, after 6 h exposure to lead caused profounder inhibitory effects on StAR protein expression and steroidogenic enzymes and then progesterone production compared to 2- or 3-h lead treatments in MA-10 mouse Leydig tumor cells. PMID- 12647777 TI - Variation of chromogranin A serum levels after radical retropubic prostatectomy for prostate adenocarcinoma. AB - This study evaluated perioperative and postoperative variations in serum CgA levels induced by radical retropubic prostatectomy (RRP) and their relationship with serum PSA levels in prostate cancer patients. Thirty consecutive patients with clinically localized adenocarcinoma of the prostate undergoing RRP were prospectively analyzed. Serum levels of CgA and total PSA were analyzed in each case preoperatively (time 0), at removal of the prostate (time 1), 1 h after the end of RRP (time 2) and then at regular postoperative intervals till 12 weeks (time 14). During the postoperative period no adjuvant therapies were performed and none of the 30 cases showed biochemical (PSA > 0.2 ng/mL) and/or clinical progression. Mean preoperative serum levels of CgA were 57 +/- 14 ng/mL. Immediately after the surgical removal of the prostate gland (time 1), in all 30 cases there was a significant (time 0-time 1: p = .001) increase in serum PSA, but a nonsignificant modification in serum CgA levels (60 +/- 15 ng/mL). After time 1, serum PSA levels progressively decreased to below the detection limit of 0.2 ng/mL. On the contrary, at time 2, serum CgA levels were postoperatively increased (time 2 = 145 +/- 47) and they remained significantly higher than preoperative values (time 0) till the 21-day postoperative interval (time 11). Moreover, at the last control (time 14) mean and median CgA levels were very similar to those shown preoperatively (time 14: 58 +/- 18 ng/mL). In patients with untreated clinically localized adenocarcinoma of the prostate submitted to RRP, surgical and postoperative stress, more than surgical manipulation of the prostate gland, could produce a significant increase in serum CgA levels maintained for a longer period when compared to the increase in serum PSA levels. PMID- 12647779 TI - Ultrastructural analysis of TESE and semen sperm tails from patients exhibiting absolute immotility on the day of ICSI. AB - Sperm flagellar pathology was found to be the underlying cause of motility disorders that lead to male infertility. Conventional in vitro fertilization (IVF) procedures will fail when sperm show a total absence of motility. In such difficult cases intracytoplasmic sperm injection (ICSI) is the only available technique to fertilize an oocyte. Fertilization rates are low and may also be reduced when immotile sperm are used for ICSI from ejaculate of other than epididiymal or testicular origin. Presence of totally immotile sperm in the ejaculate on the day of ICSI if spermatogenesis is normal testicular sperm recovery can improve ICSI outcomes. But for patients having severe morphological or functional sperm defects embryos of lower quality tend to be produced when totally immotile sperm are used. In this study the 2 patients exhibiting totally immotile sperm in their ejaculates and TESE samples on the day of ICSI showed the same ultrastructural abnormalities. Peri-axonemal and axonemal abnormalities that were seen in association with sperm nucleus structural defects suggested that the source of sperm has no effect on morphologic characteristics and also reflects abnormality in both spermatogenesis and spermiogenesis. In this study the two patients who presented with oligoteratozoospermia with total immotility, using either ejaculate or TESE sperm fertilization and embryo development, can be obtained with ICSI, but no pregnancies were established after embryo transfers. PMID- 12647778 TI - Sperm DNA fragmentation is increased in couples with unexplained recurrent pregnancy loss. AB - Previous studies have indicated that sperm quality may be related to unexplained recurrent pregnancy loss. This study evaluated the degree of sperm DNA fragmentation using the TUNEL assay on sperm from 24 couples with unexplained recurrent pregnancy loss (RPL) compared to sperm from 2 control groups: donors of known fertility and unscreened men from the general population. The percentage of sperm staining positive for DNA fragmentation was increased (p < .001) in the RPL group (38 +/- 4.2) compared to the donor (11.9 +/- 1.0) or general population (22 +/- 2.0) control groups. In the RPL group, no correlation was observed between semen quality parameters and the TUNEL data. These data indicate that some RPL patients have a significant increase of sperm DNA fragmentation, which may be causative of pregnancy loss in some patients. PMID- 12647780 TI - Effect of diluents on motility of ram sperm during storage at 5 degrees C. AB - This study examined the effects of 5 diluents--glucose-phosphate (GP), skim milk egg yolk (MY), egg yolk-citrate (YC), blood sera (S), and seawater (SW)--on daily motility of ram sperm stored at 5 degrees C for a short time. Ram semen collected by artificial vagina was diluted. Percentage of motile spermatozoa (PM) was assessed every 24 h up to day 10 after dilution. The different kinds of diluents except for SW had no effect on the primary motility of sperm and the primary motility was 0 with SW. There was a significant effect (p<.01) of diluent type and storage times in the conservation rates after cooling for PM. The GP diluent achieved greater values (80.5+/-0.88%) than S (69.5+/-1.70%) and MY (78.0+/ 1.17%), while YC showed an intermediate value (79.0+/-1.24%). In relation to time of storage, GP and YC diluents showed the PM values up to day 10, but these values were 8 and 7 days for MY and S, respectively. The GP and YC extenders maintained a PM > 50% up to day 4, while MY was greater than 50% up to day 3 and S to day 2. PMID- 12647781 TI - Clinical significance of nuclear matrix-estradiol receptor complex in human sperm. AB - Estrogen receptor levels were determined in nuclear matrix of human spermatozoa of normospermic fertile men and idiopathic infertile men. In noncapacitated sperm cells, the distribution of the estrogen receptor in normospermic fertile men was present in 50% of the cases, whereas in idiopathic infertile men was present in 19% of the cases. In capacitated sperm cells, the distribution of the estrogen receptor in normospermic fertile men was present in 80% of cases, whereas in idiopathic infertile men was present in 31% of the cases. The values of estrogen receptors in capacitated and noncapacitated sperm cells of normospermic fertile men were 91 +/- 21 and 26 +/- 7 fmol/10(8) sperm cells, respectively, whereas in capacitated and noncapacitated sperm cells of idiopathic infertile men, the estrogen receptor values were 50 +/- 17 and 22.5 +/- 9 fmol/10(8) sperm cells, respectively. The diminution of the estrogen receptor levels in the nuclear matrix could be a biochemical indicator of male factor infertility. PMID- 12647782 TI - Day-night changes in plasma melatonin levels, synaptophysin expression and ultrastructural properties of pinealocytes in developing female sheep under natural long and short photoperiods. AB - The aim of the present study was to analyze the 24-h rhythm in plasma melatonin concentration and the day-night differences in synaptophysin expresion and ultrastructural characteristics of the pinealocytes in developing female sheep. Ewes of three different ages were examined: infantile (1-6 months old), pubertal and early fertile age (9-24 months old) and adult (36-60 months old). Experiments were conducted under natural non-stimulatory (long) and stimulatory (short) photoperiods. The obtained results were similar for both analyzed photoperiods. Plasma melatonin concentration, measured in samples obtained every 4 h, showed a similar pattern in the three age groups, with peak values at 02:00 h and troughs at 14:00 h. Mean value of plasma melatonin levels in 9-24 month-old sheep was significantly greater than that in younger or older sheep. The weight of pineal glands obtained at night (02:00 h) was significantly higher than in daylight (14:00 h). Pubertal and early fertile sheep had the largest pineal glands. The pineal volume, and the total number of pinealocytes per gland of 9-24 months-old sheep differed significantly from that of younger or older sheep. The pineal volume, and the mean volume of pinealocytes was significantly greater in animals killed at night. Number of pinealocytes did not vary between animals killed during daylight or at night. The mean volumen of pinealocytes did not show statistical differences between the age groups. In quantitative ultrastructural analysis of pinealocyte cells, the relative volume of mitochondria, rough endoplasmic reticulum and Golgi complexes was significantly greater in 9-24 month old sheep and in animals killed at night. The relative volume of lipid droplets was highest in older sheep. Collectively, the data support the existence of developmental changes in pinealocyte morphology and quantity, partially in coincidence with a higher melatonin secretion rate. PMID- 12647783 TI - Beta1-integrins co-localize with Na, K-ATPase, epithelial sodium channels (ENaC) and voltage activated calcium channels (VACC) in mechanoreceptor complexes of mouse limb-bud chondrocytes. AB - Interactions between chondrocytes and their extracellular matrix are partly mediated by beta1-integrin receptors. Recent studies have shown that beta1 integrins co-localize with a variety of cytoskeletal complexes, signaling proteins and growth factor receptors. Since mechanosensitive ion channels and integrins have been proposed to participate in skeletal mechanotransduction, in this study, we investigated the possible co-localization of beta1-integrins with two ion channels and a P-type ATPase in mouse limb-bud chondrocytes. The alpha subunits of Na, K-ATPase, the epithelial sodium channel (ENaC) and the voltage activated calcium channel (VACC) were immunostained in organoid cultures derived from limb-buds of 12-day-old mice using well-characterized antibodies. Indirect immunofluorescence revealed abundant expression of beta1-integrins and each of the selected systems in limb-bud chondrocytes. Two-fluorochrome immunostaining demonstrated that beta1-integrin, Na, K-ATPase, ENaC and VACC co-localize in chondrocytes. Co-imunoprecipitation experiments revealed co-localization and association of integrins with ENaC, VACC and Na, K-ATPase. Cellular responses and signaling cascades initiated by the influx of calcium or sodium through putative mechanosensitive channels may be regulated more effectively if such channels were organized around integrins with receptors, kinases and cytoskeletal complexes clustered about them. The close proximity of ATPase ion pumps such as Na, K ATPase to chondrocyte mechanoreceptor complexes could facilitate rapid homeostatic responses to the ionic perturbations brought about by activation of mechanically gated cation channels and efficiently regulate the intracellular milieu of chondrocytes. PMID- 12647784 TI - Proliferative activity of well differentiated neuroendocrine tumours of the gut. AB - Neuroendocrine tumours of the gastrointestinal tract are relatively uncommon neoplasms with, in spite of their characteristic morphology, relatively unpredictable biological behaviour. In some sites, notably the appendix, these tumours are largely benign whereas at other localisations, such as the small bowel, metastases occur and the outcome is less favourable. Given the lack of discriminative power of histological parameters, immunohistochemical parameters have been proposed. Of these the Ki-67 index, as an indicator of proliferative activity, has shown some promise. In order to assess their proliferative activity and the potential contribution of this parameter to defining biological behaviour, we performed Ki-67 immunostaining of a series of 64 well differentiated neuroendocrine tumours of the gut (stomach, small bowel, appendix, colon and rectum). Ki-67 labeling index, based upon counting of up to 5000 cells, ranged between 0 and 6.1%. No difference was found according to age, gender, size, location or TNM classification. Ki-67 labeling index of midgut endocrine tumours of long term surviving patients did not differ from patients that died. We conclude that Ki-67 labeling index as an indicator of proliferative activity of well differentiated neuroendocrine tumours of the digestive tract does not correlate with size nor site nor stage. Even though only small numbers of tumours could be analysed, which hampered appropriate statistical analysis, it seems unlikely that proliferative activity has potential as an independent prognostic parameter for this type of tumour. PMID- 12647785 TI - Altered Cx43 expression during myocardial adaptation to acute and chronic volume overloading. AB - Gap-junctions are specialized regions of intercellular contacts allowing electrical impulse propagation among adjacent cardiomyocytes. Connexin43 (Cx43) is the predominant gap-junction protein in the working ventricular myocardium and its reduced expression has been extensively implicated in the genesis of conduction abnormalities and re-entry arrhythmia of chronically hypertrophied hearts. In contrast, data on the role played by this protein during cardiac remodeling and early phases of developing hypertrophy are lacking. Therefore, in the present study, we investigated this issue using an experimental model of pig left ventricle (LV) volume overloading consisting in the creation of an aorto cava fistula. At scheduled times (6, 24, 48, 96, 168 h, and 2, 3 months after surgery) echocardiographic and haemodynamic measurements were performed and myocardial biopsies were taken for the morphological and biochemical analyses. When faced with the increased load, pig myocardium underwent an initial period (from 6 up to 48 h) of remarkable tissue remodeling consisting in the occurrence of cardiomyocyte damage and apoptosis. After that time, the tissue developed a hypertrophic response that was associated with early dynamic changes (up regulation) in Cx43 protein expression, as demonstrated by Western blot and confocal immunofluorescence analyses. However, an initial transient increase of this protein was also found after 6 h from surgery. With the progression of LV hypertrophy (from 168 hr up to 3 months), a reduction in the myocardial Cx43 expression was, instead, observed. The increased expression of Cx43 protein during acute hypertrophic response was associated with a corresponding increase in the levels of its specific mRNA, as detected by RT-PCR. We concluded that up regulation of Cx43 gap-junction protein could represent an immediate compensatory response to support the new working conditions in the early stages of ventricular overloading. PMID- 12647786 TI - In situ visualization of o-phthalate esters in gastrointestinal tract of the frog Rana esculenta. AB - The regional distribution and relative occurrence of phthalates were studied immunohistochemically by confocal laser scanning microscopy in the alimentary tract of the green frog, Rana esculenta, using an antibody against o-phthalate esters. Many positive sites indicating the basal presence of phthalate esters were identified. The immunoreactive cells were located in the gastric glands of the stomach and in the intestinal epithelium regions with variable frequencies. The regional distribution of phathalate-accumulating cells resembled that of fish and demonstrated that these endocrine disruptors not only enter via the alimentary canal, but also bioaccumulate inside cells specialized in secretion as well as absorption functions. PMID- 12647787 TI - Effects of the proapoptotic drug prodigiosin on cell cycle-related proteins in Jurkat T cells. AB - Prodigiosin (PG) is a red pigment produced by Serratia marcescens with immunosuppressive and apoptotic activities. In this study, we sought to examine the effect of PG on cell cycle-related proteins. The antiproliferative activity of PG was tested using human Jurkat leukaemia T cells in culture. PG-inhibited cell proliferation was determined using thymidine incorporation assay. PG arrested cell cycle was analysed using immunoblot analysis with specific antibodies against cell cycle-related proteins and kinase assays of cdk2. Apoptosis was determined by Hoechst staining and analysis of DNA fragmentation. PG inhibited cyclin E, cdk2, p27 and p21, the induction of the cyclin A-cdk2 and cyclin E-cdk2 kinase activity, and the phosphorylation of Rb in leukaemic Jurkat cells. We confirmed that PG induces apoptosis by the characteristic DNA laddering pattern and condensed nuclei or apoptotic bodies identified by fluorescence microscopy. These results indicate that PG and other family members form a new group of molecules with a common mechanism of action and specific molecular targets, raising the possibility of their therapeutic use as antineoplastic drugs. PMID- 12647788 TI - Hydroxyurea (HU)-induced apoptosis in the mouse fetal tissues. AB - Hydroxyurea (HU), a ribonucleotide reductase inhibitor, induces morphological anomalies in the central nervous system (CNS), craniofacial tissues and limb buds in animals, and neonatal respiratory distress in humans. In the present study, pregnant mice were treated with 400 mg/kg of HU at day 13 of gestation, and their fetuses were examined from 1 to 48 hours after treatment (HAT) to find a clue to clarify the mechanisms of HU-induced fetotoxicity and teratogenecity. At 6 and 12 HAT, a moderate to marked increase in the number of pyknotic cells was detected in the CNS and lung. A mild increase in the number of pyknotic cells was also found in the craniofacial mesenchymal tissues, limb buds and so on. These pyknotic cells had nuclei positively stained by the TUNEL method, which is widely used for the detection of apoptotic nuclei, and they also showed electron microscopic characteristics identical to those of apoptotic cells. The present results suggest that the HU-induced fetotoxicity is characterized by excess apoptotic cell death in the fetal tissues, and that such excess cell death in the fetal CNS, lung, craniofacial tissue and limb bud may have a certain relation to the later occurrence of morphological or functional anomalies reported in these tissues following HU-administration. PMID- 12647789 TI - Distinctive expression of STAT3 in papillary thyroid carcinomas and a subset of follicular adenomas. AB - Hepatocyte growth factor (HGF), HGF receptor (c-met) and the interleukin 6 (IL-6) are expressed in thyroid nodules. In extra-thyroidal tissues, the HGF/c-met and IL-6/IL-6 receptor (IL-6R) systems activate STAT3, a member of the signal transducers and activators of transcription (STATs) family. To evaluate whether either system utilizes STAT3 in thyroid nodules, we examined the immunohistochemical expression of HGF, c-met, IL-6, IL-6R, STAT3 in 6 normal thyroids and in 68 thyroid nodules. STAT3 expression was observed in 12/12 (100%) papillary thyroid carcinomas (PTC) but in none of the follicular tumors. Among benign thyroid nodules, only 2/10 (20%) follicular adenomas (FA) were STAT3+. All these 14 STAT3+ cases expressed both HGF and c-met, but only 5 PTC co-expressed IL-6 and IL-6R and the 2 FA were IL-6+ but IL-6R-. The remaining 8 FA were all HGF/c-met-, but IL-6+; of these 8, only 2 were also IL-6R+. In conclusion, in thyroid nodules STAT3 is expressed only in PTC and a number of FA. Since these cases are consistently HGF+/c-met+ and only one-third of them co-express IL-6/IL 6R, STAT3 expression correlates with the HGF/c-met expression, not with the IL 6/IL-6R expression. The 100% rate of expression of the HGF/c-met/STAT3 signaling in PTC could be relevant for the establishment of the papillary phenotype. Because of the communeness of a HGF/c-met/STAT3 pattern between all PTC and a subset of FA, we speculate that a fraction of FA may progress to PTC. PMID- 12647790 TI - Prognostic significance of metallothionein expression in correlation with Ki-67 expression in adenocarcinomas of large intestine. AB - The study aimed at determining levels of metallothionein (MT) and Ki-67 antigen expression in adenocarcinomas of large intestine and examining relation of the expression levels with various clinical and pathological variables. The studies were performed on 81 cases of large intestine adenocarcinoma. Using immunocytochemistry, expressions of MT (positive reaction in 73 cases) and of Ki 67 (positive reaction in 79 cases) antigen were examined and the obtained results were compared with, i.a., grade (G) of the tumour and depth to which intestinal wall was infiltrated by individual tumours. Patient survival analysis was also performed, as correlated to expression levels of the two antigens. The obtained results permitted to disclose that the lower was grade of histological differentiation (G2, G3), the more pronounced was expression of MT and Ki-67. Also, the deeper was neoplastic infiltration of intestinal wall, the more pronounced was MT and Ki-67 expression. Despite the relatively strong correlation between MT expression and Ki-67 expression (r=0.536; p<0.05), only Ki-67 antigen expression in large intestine adenocarcinomas was inversely correlated to survival of the patients. Ki-67 proved to be a better prognostic marker, as compared to MT, in large intestine adenocarcinomas. PMID- 12647791 TI - Peptidergic innervation in the rat carotid body after 2, 4, and 8 weeks of hypocapnic hypoxic exposure. AB - The distribution and abundance of neuropeptide-containing nerve fibers were examined in the carotid bodies of rats exposed to hypocapnic hypoxia (10% O2 in N2) for 2, 4, and 8 weeks. The carotid bodies after 2, 4, and 8 weeks of hypoxic exposure were enlarged by 1.2-1.5 times in the short axis, and 1.3-1.7 times in the long axis in comparison with the normoxic control ones. The enlarged carotid bodies contained a number of expanded blood vessels. Mean density per unit area (10(4) microm2) of substance P (SP) and calcitonin gene-related peptide (CGRP) immunoreactive fibers was transiently high in the carotid bodies after 4 weeks of hypoxic exposure, and decreased significantly to nearly or under 50% after 8 weeks of hypoxic exposure. Density of vasoactive intestinal polypeptide (VIP) immunoreactive fibers increased significantly in all periods of hypoxic exposure observed, and was especially high in the carotid bodies after 4 weeks of hypoxic exposure. Density of neuropeptide Y immunoreactive fibers was unchanged in the carotid bodies during hypoxic exposure. These characteristic changes in the density of SP, CGRP, and VIP fibers in the carotid bodies after 4 weeks of hypoxic exposure suggest that the role of these neuropeptide-containing fibers may be different in the carotid bodies after each of three periods of hypoxic exposure, and that the peptidergic innervation after 8 weeks of hypoxic exposure may show an acclimatizing state. PMID- 12647792 TI - Postnatal differentiation of the immunohistochemical expression of aromatase P450 in the rat pituitary gland. AB - At our laboratory, we have recently demonstrated the immunohistochemical expression of aromatase P450 in the pituitary glands of adult rats; this expression was seen to be sex-dependent. In order to determine whether the changes in the expression of the enzyme are related to changes in the gonadal sphere and whether the expression of the enzyme is related to the postnatal differentiation of hypophyseal cytology, in the present work we performed an immunohistochemical study in the rat pituitary gland from birth to old age. The immunohistochemical reaction to aromatase was evident and very generalized at 7 days after birth, with no large differences between the male and female animals. At 14 days the immunohistochemical reaction was decreased in the females, with no changes in the males. At 17 days, aromatase immunoreactivity in the pituitary glands of female rats was very weak whereas the males showed large numbers of reactive cells. These observations were further pronounced at 21 days and 2 months of life. At 24 months, the immunoreactivity found in the pituitary glands of the male rats had almost completely disappeared. Our results show that a postnatal differentiation in the immunohistochemical expression of aromatase occurs; this is tightly linked to sexual activity and is lost in old age. This suggests that hypophyseal aromatase would be related to the mechanisms of action of gonadal steroids on hypophyseal differentiation and secretion. PMID- 12647793 TI - Preferential expression of cystein-rich secretory protein-3 (CRISP-3) in chronic pancreatitis. AB - BACKGROUND: Chronic pancreatitis (CP) is a progressive inflammatory process resulting in exocrine and endocrine pancreatic insufficiency in advanced stages. Cysteine-rich secretory protein (CRISP-3) has been identified as a defense associated molecule with predominant expression in the salivary gland, pancreas and prostate. AIMS: In this study, we investigated CRISP-3 expression in normal pancreatic tissues, chronic pancreatitis tissues, pancreatic cancer tissues and pancreatic cancer cell lines, as well as in other gastrointestinal organs. MATERIALS AND METHODS: 15 normal pancreatic tissues, 14 chronic pancreatitis tissues and 14 pancreatic cancer tissues as well as three pancreatic cancer cell lines were analyzed. Moreover, hepatocellular carcinoma and esophageal, stomach and colon cancers were also analyzed together with the corresponding normal controls. RESULTS: CRISP-3 was expressed at moderate to high levels in chronic pancreatitis tissues and at moderate levels in pancreatic cancer tissues but at low levels in normal pancreatic tissues, and was absent in three pancreatic cancer cell lines. CRISP-3 expression was below the level of detection in all cancerous gastrointestinal tissues and in all normal tissues except 2 of 16 colon tissue samples. CRISP-3 mRNA signals and immunoreactivity were strongly present in the cytoplasm of degenerating acinar cells and in small proliferating ductal cells in CP tissues and CP-like lesions in pancreatic cancer tissues. In contrast, CRISP-3 expression was weak to absent in the cytoplasm of cancer cells as well as in acinar cells and ductal cells in pancreatic cancer tissues and normal pancreatic tissues. CONCLUSION: These results reveal that the distribution of CRISP-3 in gastrointestinal tissues is predominantly in the pancreas. High levels of CRISP-3 in acinar cells dedifferentiating into small proliferating ductal cells in CP and CP-like lesions in pancreatic cancer suggests a role of this molecule in the pathophysiology of CP. PMID- 12647794 TI - Antitumor effect of intratumoral administration of dendritic cell combination with vincristine chemotherapy in a murine fibrosarcoma model. AB - A new antitumor therapeutic strategy utilizing the combined effect of chemotherapy and DC (dendritic cell)-based immunotherapy was designed, and the effect of intratumoral injections of unpulsed, immature DCs was evaluated after in vivo pretreatment of vincristine on tumor growth in a murine fibrosarcoma tumor model. Vincristine exerted a much more potent apoptosis/necrosis-inducing effect on MCA-102 tumor cells than on DCs both in vitro and in vivo. Moreover, CD11c, CD40, CD80 and CD86 molecules on DCs were not downregulated after treatment with vincristine either in vitro or in vivo. The growth of tumor significantly regressed in the group which received the combined vincristine chemotherapy with intratumoral administration of DCs in contrast to the untreated group, the group treated with DCs alone, and the group treated with vincristine alone. In particular, an upregulated expression of CD40, CD80 and CD86 molecules on DCs was found in the combination treatment group. Furthermore, the number of CD4+ and CD8+ T cells and the staining intensity of their CD4 and CD8 surface molecules also increased after the combination treatment. Therefore, our results indicate the feasibility of this combination therapy with vincristine chemotherapy and DC-based immunotherapy as an efficient antitumor strategy for the treatment of fibrosarcoma. PMID- 12647796 TI - Astrogliosis in different zones of the spinal cord gray matter after sciatic nerve axotomy in the newborn rat: a morphometric and immunohistochemical study. AB - Astrocytic response following unilateral sciatic nerve axotomy was examined in the spinal gray matter of newborn rats. Using an antiserum to glial fibrillary acidic protein (GFAP), immunoreactive astrocytes were studied in the ventral, dorsal and transitional region between the dorsal and ventral gray matters (TDVG) at intervals of one day, one week, two weeks and one month postaxotomy. The axotomized side showed an obvious increase in the number of immunoreactive astrocytes at one week, two weeks and one month after surgery. The numerical density per area of the glial cells (N(a)) was determined in all groups on both the intact and axotomized sides, and it increased in all groups at the axotomized sides. The percentage of glial cell increase (Pgi) was also determined. At the ventral horn Pgi increased at day one and continued to increase in all groups, while the increase in TDVG and the dorsal horn occurred at later time points. The total motoneuron count in the ventral horn at the axotomized and intact sides was done at all time points, and the percentage of motoneuron reduction (Pmr) was calculated, the highest Pmr being noticed at one month (41%). A nonlinear regression for Pmr and Pgi showed that the rate of Pgi was approximately double that of Pmr. The rate of glial cell increase at each time point (one day, one week, two weeks and one month groups) was calculated, and the highest rate of glial cell increase in the ventral horn occurred one week after axotomy, while the highest rate in the dorsal horn and TDVG occurred at the second week. The conclusion of the study is that there may be an initial post-axotomic proliferative phase of the glial cells, which was followed by a differentiation phase. Also a gradient of an increase in the rate glial cell proliferation was noticed from the ventral horn toward the dorsal horn, maybe due to stimulation by a paracrine factor. PMID- 12647795 TI - Expression of cyclin D3 and cyclin E and identification of distinct clusters of proliferation and apoptosis in diffuse large B-cell lymphomas. AB - In the present study 79 cases of de novo Diffuse Large B-cell Lymphomas (DLBCL) were studied in order: a) to analyse the expression of cyclin D3, cyclin E and cyclin D1 in relation to other proliferative features (expression of Ki67, cyclin A and cyclin B1), the apoptosis status and the expression of p53, Rb, p16 and p27; and b) to determine whether distinct clusters of proliferation and apoptosis could be identified in DLBCL. Overexpression of cyclin D3 and cyclin E was found in 35/79 (43%) and 18/79 (22%) cases, respectively, whereas overexpression of cyclin D1 was not detected in any case. In most cases (39/46) overexpression of cyclin D3 and cyclin E was mutually exclusive possibly reflecting different underlying pathways inducing deregulated expression of these cyclins. In most cases (29/35) overexpression of cyclin D3 was mutually exclusive with Rb/p16 aberrant expression status supporting an oncogenic role for cyclin D3 and suggesting that the pathogenetic effect of cyclin D3 overexpression occurs through perturbation of the Rb1 pathway. Combined alterations of the P53 and the Rb/p16/cyclin D3 expression status were significantly associated with higher mean values of cyclin A (p=0.023) and cyclin B1 (p=0.033) indicating that concurrent impairment of the p53 and Rb1 pathways induces increased tumour cell proliferation in DLBCL. Cluster analysis of the apoptosis and the proliferation status permitted separation of DLBCL into distinct groups with low (44 cases) and high (18 cases) apoptotic activity and into distinct groups with low (32 cases), intermediate (36 cases) and high (11 cases) proliferative activity. The identification of distinct clusters with respect to the proliferation and the apoptosis status indicates that groups with distinct cellular kinetic properties can be defined in the histological group of DLBCL. PMID- 12647797 TI - 65Zn uptake in the rat cerebellum and brainstem. AB - We have studied the autoradiographic uptake of 65Zn in the cerebellum and brainstem of the rat, contrasting these results with Timm's positivity in these structures. Both, autoradiographic uptake and histochemical positivity, have demonstrated Zinc in a location that could be accepted as in climbing fibres and glomeruli of the cerebellum cortex, and also in brainstem neurons that project their axons to the cerebellum cortex, suggesting a circuit where zinc may act as a neuromodulator. PMID- 12647798 TI - In vitro acute toxicity of anionic surfactant linear alkylbenzene sulphonate (LAS) on the motility of gilthead (Sparus aurata L.) sperm. AB - This paper describes the acute toxicity of a known anionic surfactant, Linear Alkylbenzene Sulphonate (LAS), on the quality of gilthead Sparus aurata L. sperm. The parameter used to judge exposure effectiveness was sperm motility as well as its fertilizing ability after being combined with unexposed gilthead eggs. Preincubation of sperm suspensions with concentrations of LAS of 0.1, 0.5, 1, 2 and 4 mg/L caused decrease in sperm motility and fertilizing ability. In this respect, percentages of motile sperm were respectively 89.8+/-9.8, 81.7+/-16.3, 69.5+/-21.3, 57.1+/-19.1 and 21.2+/-10.9%. With regard to the percentage of fertilization success, the results were 85.7+/-8.1, 75.1+/-20.2, 62.9+/-19.7, 52.7+/-19.2 and 14.2+/-7.9% respectively. At concentrations of LAS of 0.5 mg/L or higher, the differences in both percentage of motility and fertilizing ability with controls were significant (p<5%). Although extrapolation from the laboratory to the field requires caution, the results of this work demonstrated that low level surfactant pollution may impact directly on reproduction of the free gametes (sperm) released into water. It may lead to a long-term decline and eventual extinction of gilthead populations in nature when they are located close to effluents that are either untreated or receive inadequate secondary treatment. It is also quite important because this species constitutes an important link in the food chain and its death via exposure to surfactants may imbalance the littoral ecosystem. PMID- 12647799 TI - Autofluorescence spectroscopy of malpighian epithelial cells, as a new tool for analysis of cervical cancer precursors. AB - A spectroscopic analysis of autofluorescence was investigated within the cell cytoplasm from cervical malpighian epithelia prepared on Thin-Prep smears. Autofluorescence emission spectra from 22 cervix were analyzed by microspectrofluorometry under a 363 nm laser excitation. Among the analyzed cervix, 6 were in normal limits, 6 in inflammatory limits, 5 were evocative of Low-Grade Squamous Intraepithelial Lesions (LGSILs) and 5 were evocative of High Grade Squamous Intraepithelial Lesions (HGSILs). Cytoplasmic emission intensities at 450 nm of cells from inflammatory, LGSIL and HGSIL cervix were equivalent and were 3-fold higher than from normal cervix. All smears presented a two-fold lower autofluorescence emission in the cytoplasm than in the nucleus. The spectral profile analysis allows the discrimination of cells from inflammatory, LGSIL and HGSIL cervix. The 525/425 nm emission ratios were 0.75+/-0.1, 0.96+/-0.04 and 1.2+/-0.1 for inflammatory, LGSIL and HGSIL, respectively. We suggest that smears of normal, inflammatory, LGSIL and HGSIL cervix could be discriminated by the analysis of the 450 nm emission intensity and 525/425 nm emission ratios from cells of malpighian epithelia. PMID- 12647800 TI - Review of papillary renal cell carcinoma with focus on clinical and pathobiological aspects. AB - Recent studies have shown that papillary renal cell carcinoma (RCC) is clinically and genotypically a distinct entity. Papillary RCCs account for about 10-15% of renal parenchymal neoplasms. Macroscopically, the cut surface is yellow or brown in color and large tumors frequently show cystic change. Hemorrhage and necrosis are common. Histologically, Delahunt and Eble have classified papillary RCCs into type 1 (small cells, single layer) and type 2 (large cells, pseudostratification) according to the cytoplasmic volume and thickness of the lining cells. In chromosomal analysis, gain of chromosomes 7 and 17, loss of Y chromosome and additional gains (chromosome 3q, 8p, 12q, 16q and 20q) are frequently found in type 1 papillary RCCs, but the chromosomal aberration of type 2 papillary RCCs seems to be more heterogenous than that of type 1 papillary RCCs. Mutations of MET proto-oncogenes in some cases of both hereditary and sporadic papillary RCCs have recently been detected. Furthermore, all hereditary and sporadic papillary RCCs with MET proto-oncogene show type 1 histological features. Type 1 papillary RCCs generally seem to have a favorable prognosis, but type 2 tumors have a worse prognosis than do type 1 tumors. Papillary RCCs with involvement of the X chromosome and cancer syndrome with predisposition to cutaneous/uterine leiomyomas and papillary RCCs, the histological features of which are basically different from those of usual papillary RCCs, have also been recently reported. Since papillary RCCs seem to constitute clinically, histologically, and even genetically more heterogenous groups than previously thought, further investigations are needed to characterize the subtype of papillary RCC. PMID- 12647801 TI - Suicide gene therapy with Herpes simplex virus thymidine kinase and ganciclovir is enhanced with connexins to improve gap junctions and bystander effects. AB - Connexins are proteins that form gap junctions between cells in various mammalian tissues. Because of their role in intercellular communication, connexins are important in the bystander cell death seen in Herpes simplex virus-thymidine kinase (HSV-TK) gene therapy for brain tumors. A selective review of connexin transduction/transfection studies with particular emphasis to central nervous system tumor cells is presented. In addition, specific references to studies with cell types that demonstrate low gap junction intercellular communication are presented. Data are included with the HT-29 colorectal tumor cell line to support the concept that enhancing gap junction protein expression in otherwise low gap junction communicating HT-29 cells increases bystander cell death and reduces tumor burden beyond what might be expected from HSV-TK and ganciclovir (GCV) treatment alone. Maximum in vitro bystander cell death was always produced when GCV treated co-cultures of TK-transduced and non-TK-transduced HT-29 cell lines were also transduced with connexin-43. When connexin was present in only one group of cells in the co-culture, there was more bystander cell death observed with connexin transduced into the non-TK-transduced cells, rather than the TK transduced cells. The data presented reinforces conclusions made from earlier findings from cell line mixing experiments in which the non-TK-transduced cell population determined the level of bystander cell death (Burrows et al., 2002). PMID- 12647802 TI - Ubiquitinated inclusions and neuronal cell death. AB - Ubiquitinated inclusions and selective neuronal cell death are considered the pathological hallmarks of Parkinson's disease and other neurodegenerative diseases. Recent genetic, pathological and biochemical evidence suggests that dysfunction of ubiquitin-dependent protein degradation by the proteasome might be a contributing, if not initiating factor in the pathogenesis of these diseases. In neuronal cell culture models inhibition of the proteasome leads to cell death and formation of fibrillar ubiquitin and alpha-synuclein-positive inclusions, thus modeling some aspects of Lewy body diseases. The processes of inclusion formation and neuronal cell death share some common mechanisms, but can also be dissociated at a certain level. PMID- 12647803 TI - Selection events in directing B cell development. AB - Homeostasis in the B cell compartment (as well as in T cells) is controlled by tightly regulated selection events. Throughout their life span, B cells are subjected to selection signals determining not only developmental progression, but also maturation and survival. It is now clear that most of these signals require the expression of B cell antigen receptor (or preB receptor) with functional signaling capacity. The administration of numerous mutations into the mouse germline enabled us to identify several checkpoints along the B cell developmental pathway, and provided us with powerful experimental tools to probe for selection events regulating developmental progression. In here, we will discuss recent studies in this field. PMID- 12647805 TI - SV40 early region oncoproteins and human cell transformation. AB - We now understand neoplastic transformation to be the consequence of multiple acquired genetic alterations. The combination of these acquired changes confer the various phenotypes that constitute the clinical features of cancer. Although only rare human cancers derive from a viral etiology, the study of DNA tumor viruses that transform rodent and human cells has led to a greater understanding of the molecular events that program the malignant state. In particular, investigation of the viral oncoproteins specified by the Simian Virus 40 Early Region (SV40 ER) has revealed critical host cell pathways, whose perturbation play an essential role in the experimental transformation of mammalian cells. Recent work has re-investigated the roles of two SV40 ER oncoproteins, the large T antigen (LT) and the small t antigen (ST), in human cell transformation. Co expression of these two oncoproteins, together with the telomerase catalytic subunit, hTERT, and an oncogenic version of the H-Ras oncoprotein, suffices to transform human cells. LT inactivates two key tumor suppressor pathways by binding to the retinoblastoma protein (pRB) and p53. The ability of ST to transform human cells requires interactions with PP2A, an abundant family of serine-threonine phosphatases. Here we review recent developments in our understanding of how these two viral oncoproteins facilitate human cell transformation. PMID- 12647804 TI - Aberrant PAX3 and PAX7 expression. A link to the metastatic potential of embryonal rhabdomyosarcoma and cutaneous malignant melanoma? AB - Transcription factors encoded by PAX3 and PAX7 are amongst the first expressed in the embryo, being principal regulators of neurogenic and myogenic progenitor cell specification and embryonic segmentation. The basis for this review lies in the supposition that genetic programs for cell migration, thought regulated by PAX3 and PAX7 during embryonic development, become tools used by the metastatic cell. In highly metastatic neoplasms arising from cells of neurogenic and myogenic lineages such as embryonal rhabdomyosarcoma and cutaneous malignant melanoma, markedly high expression levels of PAX3 and PAX7 support this supposition. As PAX3 and PAX7 are known to play a role in the regulation of migratory events in embryogenesis, it is possible that the metastatic potential of these tumours is directly linked to migratory properties conferred them through PAX expression. Here we provide a novel perspective by correlating metastasis with expression of PAX3, PAX7 and ephrin/Eph receptors as well as NCAMs, cell surface markers normally involved in migration and adhesion during development, and propose a role for PAX genes in the increased metastatic potential of these tumours. PMID- 12647806 TI - Review of sarcomatoid renal cell carcinoma with focus on clinical and pathobiological aspects. AB - In sarcomatoid renal cell carcinoma (RCC), it is generally accepted that the sarcomatoid portion is derived from metaplastic transformation of carcinoma. Sarcomatoid RCCs account for about 1-8% of all renal tumors. Macroscopically, tumors generally form encapsulated masses and show invasive growth. Sarcomatoid RCCs originate from all subtypes of RCCs, including conventional, papillary, chromophobe, and collecting duct carcinomas. With regard to the growth pattern of the sarcomatoid component, malignant fibrous histiocytomatous, fibrosarcomatous and unclassified sarcomatous patterns are frequently seen. Immunohistochemically, sarcomatoid RCCs are generally positive for AE1/AE3, epithelial membrane antigen (EMA) and vimentin and negative for desmin, actin and S-100. Little is know about genetic alterations in sarcomatoid RCCs. Further studies are therefore needed to identify the key gene involved in sarcomatoid transformation of RCCs. PMID- 12647807 TI - Dynamics of lineage-restricted mixed chimerism following sex-mismatched allogeneic bone marrow transplantation. AB - Scant knowledge is available about the dynamics of lineage-specific mixed chimerism (Ch) following bone marrow transplantation (BMT). This review is focused on findings derived from bone marrow (BM) biopsies in patients with chronic myeloid leukemia (CML) including a sex-mismatched host/donor constellation. Appropriate techniques involved immunophenotyping by monoclonal antibodies to identify the various cell lineages, dual color fluorescence in situ hybridization (FISH) with x- and y-chromosome-specific DNA-probes and a proper detection system for a simultaneous labeling of the bcr/abl locus. A significant degree of Ch with more than 20% host CD34+ progenitors was found in the early and late (up to 200 days after BMT) posttransplant period. However, only 10% of these cells harbored the bcr/abl translocation gene. This result fits well with corresponding molecular biological findings of so-called minimal residual disease. Conversion of Ch evolved during leukemic relapse with 90% host progenitors of which 50% revealed the bcr/abl locus. A Ch of nucleated erythroid percursors (5%) and CD68+ macrophages (8%) was expressed to a significantly lower degree. The slightly increased frequency found in CD61+ megakaryocytes (16%) was probably due to the polyploid state of these cells. Similar to the CD34+ progenitor cells abrupt changes from donor to host type was associated with an insidious transformation into recurrent leukemia. The CD34+ endothelial cells showed a minor degree of Ch, because donor-derived elements ranged from 18% to 25%. Leukemic relapse was characterized by an almost complete conversion of the endothelial cells to a host type. These findings point towards a CD34+ progenitor cell origin of the (leukemic) endothelial cell layer and suggests that their dysfunction may contribute to an expansion of the neoplastic clone. PMID- 12647808 TI - Sympathetic sprouting in dorsal root ganglia (DRG): a recent histological finding? AB - During the nineties it was described, as an original finding, the existence of afferent amyelinic nerve endings in animal dorsal root ganglia (DRG) caused by diverse experimental lesions. These works do not take into account the historical studies carried out by Ehrlich (1886), Ramon y Cajal (1890) and Dogiel (1885) among others. Ramon y Cajal (1899) confirmed the existence of these nerve endings naming them after their discoverer as "Dogiel's arborisations". Ramon y Cajal claims that these endings originate from fibres of sympathetic nature, something supported by later authors devoted to this topic. In any case, the same authors remarked already a possible relationship with pathological phenomena, nonetheless always referring to the frequent occasions in which the same images appeared in healthy animals. In this work we review the bibliography about the classically named "Terminal Dogiel's nests" which in modern literature have been referred to as sprouting of sympathetic axons in dorsal root ganglia likely related with sympathetically maintained pain. Furthermore, we present the finding, not described up to date, of multiple afferent amyelinic nervous endings related with the "Terminal Dogiel's nests" observed in different DRG from young adult healthy rabbits. PMID- 12647809 TI - Leukotriene synthesis by epithelial cells. AB - Leukotrienes (LTs) are intercellular signaling molecules that evoke a variety of responses. They are best known as potent promoters of inflammation. Normally, LTs are produced primarily by leukocytes. As a result, current models regarding the production of LTs in the context of disease focus on the leukocytes as the site of production. Structural cells, including epithelial cells, are typically relegated to supportive roles. It is recognized that epithelial cells normally contain all the components necessary for LT synthesis except the enzyme 5 lipoxygenase (5-LO). There is accumulating evidence that some populations of epithelial cells normally express low levels of 5-LO and can synthesize LTs autonomously. Moreover, certain factors, including bacterial and viral infection, can promote the expression of 5-LO in airway, gastrointestinal and skin epithelial cells. The appearance of active 5-LO enzyme in epithelial cells at these sites may contribute to diseases like cancer, colitis and psoriasis. This paper reviews the state of our knowledge regarding the expression of 5-LO in epithelial cells, the factors that modify that expression, and the implications regarding pathogenesis. PMID- 12647810 TI - Apocrine secretory mechanism: recent findings and unresolved problems. AB - Cell secretion is an important physiological process that ensures smooth metabolic activities, tissue repair and growth and immunological functions in the body. It occurs when the intracellular secretory materials are released to the exterior; these may be in the form of lipids, protein or mucous and may travel through a duct system or via blood to reach the target organ. To date three types of secretory mechanisms have been characterized, they include apocrine, holocrine and exocytosis. Apocrine secretion occurs when the release of secretory materials is accompanied with loss of part of cytoplasm. The secretory materials may be contained in the secretory vesicles or dissolved in the cytoplasm that is lost during secretion. In holocrine secretion, the entire cell is secreted into the glandular lumen, and it is presumed that the intended secretory materials are contained in the cell cytoplasm. Exocytosis is the most commonly occurring type of secretion; here the secretory materials are contained in the secretory vesicles and released without loss of cytoplasm. Apocrine secretory mechanisms have not been properly discussed; for example the biochemical and physiological pathways that regulate apocrine secretory process are not clearly known. Similarly, the plasma membrane dynamics during apocrine secretion has not been researched. In other glands morphological features during apocrine secretion have not been documented. The current paper reviews what is known about apocrine secretion, recent findings and highlights on the unresolved areas for future research. PMID- 12647811 TI - Use of laser scanning cytometry to study tumor microenvironment. AB - The study of phenomena occurring in the tumor microenvironment is a challenging task because of technical difficulties, particularly when dealing with hypocellular specimens. Laser scanning cytometry (LSC) is a new laboratory technology that has been recently introduced to overcome the limitations of other traditional technologies. By combining the properties and the advantages of flow cytometry (FC) and immunohistochemistry (IHC), LSC allows the investigator to obtain objective information on DNA content, protein expression and cellular localization is combination with morphological features. It has been already shown that LSC results are reliable compared to more traditional technologies, and its implementation in the clinical routine is under way. Its use in oncology, which is rapidly expanding, spans from apoptosis analysis to DNA content quantitation and tumor cell phenotyping. Here we describe the technology underlying this novel fluorescence-based device, review its use in oncology by dissecting the phenomena occurring in the tumor microenvironment and propose its application for the immunological follow-up of malignant lesions undergoing immunotherapeutic manipulation. PMID- 12647813 TI - TrkB receptor signaling and activity-dependent inhibitory synaptogenesis. AB - When mouse organotypic cerebellar cultures were exposed to anti-GABA agents that increased neuronal activity early in development, there was a doubling of the ratio of inhibitory axosomatic synapse profiles to Purkinje cell somatic profiles after two weeks in vitro, which correlated with a decrease in spontaneous cortical discharges. When similar cultures were maintained in medium with activity blocking agents, Purkinje cell axosomatic synapses were reduced to approximately half of control values and, after recovery from activity blockade, the cultures discharged hyperactively. By contrast, the full complement of excitatory cortical synapses developed in the absence of neuronal activity. These results support the concept that neuronal activity is necessary for the complete development of inhibitory circuitry. When cerebellar cultures were simultaneously exposed to activity blocking agents and to neurotrophins BDNF or NT-4, both of which bound to the TrkB receptor, the numbers of inhibitory Purkinje cell axosomatic synapses were similar to those of untreated control cultures, and control rates of spontaneous cortical discharges were recorded. The TrkC receptor ligand, NT-3, did not promote inhibitory synapse development in the absence of neuronal activity, and such cultures exhibited hyperactive cortical discharges. These results are consistent with a role for TrkB receptor ligands in activity dependent inhibitory synaptogenesis. Subsequent exposure of cerebellar cultures to antibody to the extracellular domain of TrkB induced an increased development of Purkinje cell axosomatic synapses, while similar antibody activation of TrkC had no effect on inhibitory synaptogenesis. The promotion of inhibitory synapse development by specific antibody activation of TrkB supports the concept that signaling for activity-dependent inhibitory synaptogenesis is via the TrkB receptor. PMID- 12647812 TI - Structure and molecular organization of dendritic spines. AB - Dendritic spines mediate most excitatory synapses in the CNS and are therefore likely to be of major importance for neural processing. We review the structural aspects of dendritic spines, with particular emphasis on recent advances in the characterization of their molecular components. Spine morphology is very diverse and spine size is correlated with the strength of the synaptic transmission. In addition, the spine neck biochemically isolates individual synapses. Therefore, spine morphology directly reflects its function. A large number of molecules have been described in spines, involving several biochemical families. Considering the small size of a spine, the variety of molecules found is astounding, suggesting that spines are paramount examples of biological nanotechnology. Single-molecular studies appear necessary for future progress. The purpose of this rich molecular diversity is still mysterious but endows synapses with a diverse and flexible biochemical machinery. PMID- 12647814 TI - The pathological basis of myocardial hibernation. AB - Myocardial hibernation refers to a state of persistent regional ventricular dysfunction, in patients with coronary artery disease that is reversible with revascularization. It is part of the spectrum of pathophysiological responses to myocardial ischemia and is a particularly important concept in understanding the development and progression of ischemic cardiomyopathy. Hibernating myocardium may be associated with chronic hypoperfusion, or result from repetitive episodes of ischemia with a cumulative effect on contractile function. Mechanistic studies on myocardial hibernation have been hampered by the difficulty in developing a reproducible and reliable animal model. This review describes the pathologic changes found in hibernating myocardial segments discussing the potential mechanisms involved in their development. Depletion of cardiomyocyte contractile elements, loss of myofilaments and disorganization of cytoskeletal proteins are among the most consistently reported morphological alterations found in hibernating myocardial segments. In addition, the cardiac intersitium exhibits inflammatory changes, leading to fibrotic remodeling. Induction of cytokines and chemokines suggests an active continuous inflammatory process leading to fibrosis and dysfunction. Although, the initial response may be adaptive to ischemia, if timely revascularization is not performed, irreversible tissue injury, fibrosis and myocyte degeneration may develop. Understanding the role of inflammatory mediators in the development and progression of the cardiomyopathic process may lead to the development of specific therapeutic strategies aiming at preventing irreversible fibrosis and dysfunction. PMID- 12647815 TI - Pathogenetic mechanisms of nuclear pleomorphism of tumour cells based on the mutator phenotype theory of carcinogenesis. AB - The nuclei of the cells of most solid tumours in histopathologic preparations vary in size, shape and chromatin pattern, both from normal nuclei and from each other. These features have not been explained in terms of conventional concepts of nuclear structure and theories of carcinogenesis. In recent years, the unfolded chromosomes have been shown to occupy "domains" in the nucleus during interphase, providing a relatively uniform density of fine chromatin fibres throughout the nucleus in the living state. This is in contrast to the appearances of interphase chromatin existing as coarse clumps and fibres (heterochromatin and euchromatin respectively) as are seen in histologic preparations. Additionally, the binding of chromatin to nuclear membrane, the possible existence of a nuclear matrix, the functions of nuclear pores, and the attachments of cytoskeletal structures to the outer nuclear membrane are now recognised. Studies of genetic instability of cancer cells (many random mutations are present in the genome, which vary from nucleus-to-nucleus in individual tumours) have shown that this phenomenon occurs early in tumour formation, can be present in morphologically-normal cells adjacent to tumours, and can result in thousands of genomic events per tumour cell. These observations form the basis for the mutator phenotype/clonal selection theory of carcinogenesis, which proposes that genetic instability is an essential early part of carcinogenesis. Genetic instability has been used to explain significant cell-to-cell variability of behaviour (tumour cell heterogeneity) among cells of individual tumours. This paper proposes that a high incidence of nucleus-to-nucleus-variable mutation of the genes for factors controlling nuclear morphology in tumours can explain nucleus-to-nucleus variations of histopathologic appearance of these nuclei when some additional effects of histological processing are taken into account. PMID- 12647816 TI - Epigenetic inactivation of the Ras-association domain family 1 (RASSF1A) gene and its function in human carcinogenesis. AB - The Ras GTPases are a superfamily of molecular switches that regulate cellular proliferation and apoptosis in response to extra-cellular signals. The regulation of these pathways depends on the interaction of the GTPases with specific effectors. Recently, we have cloned and characterized a novel gene encoding a putative Ras effector: the Ras-association domain family 1 (RASSF1) gene. The RASSF1 gene is located in the chromosomal segment of 3p21.3. The high allelic loss in a variety of cancers suggested a crucial role of this region in tumorigenesis. At least two forms of RASSF1 are present in normal human cells. The RASSF1A isoform is highly epigenetically inactivated in lung, breast, ovarian, kidney, prostate, thyroid and several other carcinomas. Re-expression of RASSF1A reduced the growth of human cancer cells supporting a role for RASSF1 as a tumor suppressor gene. RASSF1A inactivation and K-ras activation are mutually exclusive events in the development of certain carcinomas. This observation could further pinpoint the function of RASSF1A as a negative effector of Ras in a pro apoptotic signaling pathway. In malignant mesothelioma and gastric cancer RASSF1A methylation is associated with virus infection of SV40 and EBV, respectively, and suggests a causal relationship between viral infection and progressive RASSF1A methylation in carcinogenesis. Furthermore, a significant correlation between RASSF1A methylation and impaired lung cancer patient survival was reported, and RASSF1A silencing was correlated with several parameters of poor prognosis and advanced tumor stage (e.g. poor differentiation, aggressiveness, and invasion). Thus, RASSF1A methylation could serve as a useful marker for the prognosis of cancer patients and could become important in early detection of cancer. PMID- 12647818 TI - Solid-phase microextraction from small volumes of sample in a glass capillary. AB - A new sampling method is proposed for solid-phase microextraction (SPME), in which the extraction is carried out in a glass capillary containing a few microliters of sample. When an adsorption-type fiber is used for SPME, the equilibrium between aqueous sample and coating can be described by a Langmuir isotherm. Since the total amount of analytes and coexisting substances stays at a low level in a small volume of sample, the linear concentration range of analytes will be extended for SPME to be applied in quantification and the interference caused by sample matrix will be reduced. In addition, sampling in a capillary has a short diffusion distance and extraction equilibrium is established in 5-10 min. It is important in clinical analysis and therapeutic drug monitoring to be able to analyse sample volumes of samples. The feasibility of the new sampling method is demonstrated by the extractions of p-hydroxybenzaldehyde and a synthetic solution containing 1-naphthol, paeonol and 1-naphthylamine. PMID- 12647819 TI - Solid-phase clean-up in the liquid chromatographic determination of polycyclic aromatic hydrocarbons in edible oils. AB - A solid-phase extraction (SPE) method for sample clean-up, followed by reversed phase high-performance liquid chromatography (HPLC) with fluorescence detection is reported for the determination of polycyclic aromatic hydrocarbons (PAHs) in edible oils. The effects of experimental variables, such as washing and elution solvents, sample solvent and drying time have been studied using C18 cartridges. Recoveries and selectivity using other sorbent materials (C8, C2, CH, PH and NH2) were also examined, with C18 being the best one. The recoveries ranged between 50 and 103% depending on the molecular mass of the PAH. The limits of quantitation were lower than 1 ng/g for most PAHs and good precision was achieved. The method was validated using certified reference materials. PMID- 12647817 TI - Current and prospective applications of metal ion-protein binding. AB - Since immobilized metal ion affinity chromatography (IMAC) was first introduced, several variants of this method and many other metal affinity-based techniques have been devised. IMAC quickly established itself as a highly reliable purification procedure, showing rapid expansion in the number of preparative and analytical applications while not remaining confined to protein separation. It was soon applied to protein refolding (matrix-assisted refolding), evaluation of protein folding status, protein surface topography studies and biosensor development. In this review, applications in protein processing are described of IMAC as well as other metal affinity-based technologies. PMID- 12647820 TI - Trace analysis of technical nonylphenol, bisphenol A and 17alpha-ethinylestradiol in wastewater using solid-phase microextraction and gas chromatography-mass spectrometry. AB - To enable high sample throughput, an automated solid-phase microextraction (SPME) method coupled with GC-MS for the trace analysis of technical nonylphenol, bisphenol A and 17alpha-ethinylestradiol was developed. The extraction performance of different SPME fibre coatings was examined, with polyacrylate proving most suitable. Although study of the extraction time showed that the analytes have not reached equilibrium after 3 h, as a compromise an extraction time of 1 h was applied in all the experiments with detection limits between 0.04 and 1 microg l(-1) for wastewater effluent. The mean reproducibility of the technique is 8% RSD. Carry-over effects are negligible. The linearity of calibration curves ranges over three orders of magnitude. The method was tested for determining the analytes in influents and effluents of constructed wetland plants and in model wastewater used in laboratory experiments. PMID- 12647821 TI - Application of the reversed-phase liquid chromatographic model to describe the retention behaviour of polydisperse macromolecules in gradient and isocratic liquid chromatography. AB - This paper illustrates how conventional models of chromatographic behaviour can be used to predict the separation behaviour of polydisperse macromolecules. Using polystyrene and polymethylmethacrylate homo- and co-polymeric standards, the models were validated by comparing experimental retention behaviour with that predicted by the chromatographic model. The experimental retention time of each of the samples was entered into a spreadsheet application, which calculated the parameters that best described retention (for a given model). When a correlation between the relevant parameters and molecular mass was established, that correlation was used to predict the change in retention behaviour over the molecular-mass range. An expression introduced in a previous paper, to calculate the critical mobile-phase composition of a homopolymer was validated using polystyrene homopolymers. A second expression, which can predict the elution behaviour of copolymers, was also validated. This expression can predict the retention of a copolymer, based solely onthe retention of the homopolymeric units that make up the copolymer. PMID- 12647822 TI - Elution behavior of polyethylene in polar mobile phases on a non-polar sorbent. AB - Linear polyethylene standards in the range of 1-500 kg/mol, dissolved in 1,2,4 trichlorobenzene, were injected into a column packed with oligo(dimethylsiloxane) modified silica gel. Fifteen polar solvents (cyclohexanone, cyclohexylacetate, cyclohexanol, nonylalcohol, dimethylformamide, dimethyl sulfoxide, ethylene- and diethylene glycol monobutyl ether, benzylalcohol, hexylacetate, bis(2-ethyl hexyl)phthalate, N,N-dimethylacetamide, propylene carbonate, dipropylene glycol and N-methyl-pyrrolidone) were evaluated as mobile phases. Depending on the type of mobile phase evaluated, different elution behaviors are observed for polyethylene: (1) polyethylene was eluted in the size exclusion mode, (2) polyethylene was eluted together with the sample solvent peak at constant elution volume, (3) polyethylene was partially or fully retained on the column. The retained polymer was easily removed from the column by injecting a small volume of trichlorobenzene. The use of ethylene glycol monobutyl ether as the mobile phase enabled separation of the polyethylene from polypropylene. In this case polypropylene is eluted in the size exclusion mode, while polyethylene is eluted at a constant elution volume or remains in the column. PMID- 12647824 TI - Isolation and identification of four flavonoid constituents from the seeds of Oroxylum indicum by high-speed counter-current chromatography. AB - Four flavonoids, chrysin, baicalein, baicalein-7-O-glucoside, baicalein-7-O diglucoside (Oroxylin B) and one unknown flavonoid have been isolated and purified for the first time in the seeds of Oroxylum indicum by high-speed counter-current chromatography with a two-phase solvent system composed of chloroform-methanol-water (8:10:5, v/v). Then, 50 mg baicalein-7-O-glucoside, 10.5 mg baicalein-7-O-diglucoside, 4.5 mg chrysin-7-O-diglucoside, 25 mg baicalein and 45 mg chrysin could be obtained after injecting 20 mg/ml sample extract ten times and their purities were 96, 90, 85, 95 and 98%, respectively. All these constituents were identified by high-performance liquid chromatography mass spectrometry and nuclear magnetic resonance. PMID- 12647823 TI - Determining orthogonal chromatographic systems prior to the development of methods to characterise impurities in drug substances. AB - To define starting conditions for the development of methods to separate impurities from the active substance and from each other in drugs with an unknown impurity profile, the parallel application of generic orthogonal chromatographic systems could be useful. The possibilities to define orthogonal chromatographic systems were examined by calculation of the correlation coefficients between retention factors k for a set of 68 drugs on 11 systems, by visual evaluation of the selectivity differences, by using principal component analysis, by drawing color maps and evaluating dendrograms. A zirconia-based stationary phase coated with a polybutadiene (PBD) polymer and three silica-based phases (base deactivated, polar-embedded and monolithic) were used. Besides the stationary phase, the influence of pH and of organic modifier, on the selectivity of a system were evaluated. The dendrograms of hierarchical clusters were found good aids to assess orthogonality of chromatographic systems. The PBD-zirconia phase/methanol/pH 2.5 system is found most orthogonal towards several silica based systems, e.g. a base-deactivated C16 -amide silica/methanol/pH 2.5 system. The orthogonality was validated using cross-validation, and two other validation sets, i.e. a set of non-ionizable solutes and a mixture of a drug and its impurities. PMID- 12647825 TI - Gradient reversed-phase liquid chromatographic-electrospray ionization mass spectrometric method for the comparison of smokeless powders. AB - A gradient reversed-phase liquid chromatographic-electrospray ionization mass spectrometric (LC-ESIMS) method was developed to determine compositional variation in the organic additives of smokeless powders. The method was optimized for the separation and detection of selected powder constituents, including diphenylamine, along with isomers of its nitroso and nitro derivatives, centralite I and II, in addition to dialkylphthalate acid esters. A series of commercially available smokeless powders was prepared by organic liquid extraction and characterized using the LC-ESIMS method. The results demonstrate the differentiation of smokeless powders by their additive profile. PMID- 12647826 TI - Using temperature gradient gas chromatography to determine or predict vapor pressures and linear solvation energy relationship parameters of highly boiling organic compounds. AB - An isothermal chromatographic method allowing determination of sigmabetaH2 and sigmaalphaH2 descriptors of the linear solvation energy relationship (LSER) was tested and results obtained are presented. This method is based on the use of four stationary phases of various polarity. On the other hand, it was demonstrated that the temperature gradient chromatography may be successfully used to determine LSER descriptors. Results of piH2, sigmabetaH2 and log L16 determination are reported. This approach opens new possibilities of precise and rapid determination of LSER descriptors of high boiling compounds using a small number of phases. It was demonstrated that the log L16 descriptor may be used to estimate vapor pressures of high boiling organic compounds with a better accuracy than those usually obtained with chromatographic methods. PMID- 12647827 TI - Analysis of low-molecular-mass organic acids using capillary zone electrophoresis electrospray ionization mass spectrometry. AB - A capillary zone electrophoresis-electrospray ionization-mass spectrometry (CZE ESI-MS) method was developed to facilitate identification and determination of eleven low-molecular-mass (LMW) organic acids (i.e. oxalic, lactic, malonic, maleic, citric, tartaric, adipic, glutaric, gluconic, isosaccharinic and succinic acid) in different sample matrices. This CZE method was adapted to suit MS conditions. Sheath liquid, sheath flow and MS parameters were optimized to achieve high mass spectrometric sensitivity. The CZE-ESI-MS procedure showed good sensitivity (limit of detection of < 0.05-0.7 mg/l for all acids), linearity (r2 = 0.9925-0.9998) and reproducibility (2.09-5.34% RSD). The applicability of the CZE-ESI-MS was demonstrated on LMW organic acids in an ale sample. In addition the (here presented) method also provided quantification of fumaric, galacturonic and 2-ketoglutaric acid with high sensitivity. PMID- 12647828 TI - Quantitation of the enantiomers of ofloxacin by capillary electrophoresis in the parts per billion concentration range for in vitro drug absorption studies. AB - Ofloxacin, a chiral fluoroquinolone, possesses two optical isomers. The antibacterial activity of S-(-)-ofloxacin is reported to be 8-128 times higher than that of R-(+)-ofloxacin. A capillary zone electrophoresis method has been developed to quantify the enantiomers of ofloxacin in high diluted samples (20 700 ng/ml for each enantiomer). After fluid-fluid extraction of ofloxacin from physiological solution electrokinetic injection was employed to improve the sensitivity. The method was optimised using a central composite design. Four experimental factors were investigated: the background electrolyte concentration, the methyl-beta-cyclodextrin concentration, the buffer pH and the temperature. The amount migrated into the capillary, determined by the peak area, the resolution between the ofloxacin enantiomers, the migration time and the generated current were evaluated as responses. The quantification limit is 11.4 ng/ml for S-ofloxacin and 10.8 ng/ml for R-ofloxacin. The method has shown good validation data in terms of precision and recovery rate. PMID- 12647829 TI - Determination of major phenolic compounds in water by reversed-phase liquid chromatography after pre-column derivatization with benzoyl chloride. AB - A simple reversed-phase LC method capable of detecting ng/ml quantities of phenolic compounds in water is described. Pre-column derivatization with benzoyl chloride is used for the separation and determination o-cresol, m-cresol, p cresol, phenol, resorcinol, catechol and hydroquinone in water. The benzoyl derivatives formed within in 15 min, were extracted with dietyl ether, and then analyzed by liquid chromatography with UV detection at 232 nm. With a mobile phase of acetonitrile-tetrahydrofuran-water (54:6:40, v/v) the seven derivatives were eluted in 15 min. The detection limits were between 0.05 and 0.50 ng/ml for 50 ml of a standard water sample. The method was applied to the analysis of phenols in wine and river water. The recovery of the derivatives from pure water was 81-94% with relative standard deviations of 2.5-5.0%. PMID- 12647830 TI - Determination of hexavalent chromium by on-line dialysis ion chromatography in a matrix of strong colourants and trivalent chromium. AB - Hexavalent chromium detection in the presence of a high load of colourants without any false positive and in-procedure oxidation of Cr(III) is an important area of study. Colourants are a class of interfering substances in many spectroscopic analyses and chromatographic separations and detection. A purification method using an on-line dialysis technique for ion chromatography (IC) has been developed to remove water-soluble anionic dyes and particulate colourants and other substances to facilitate Cr(VI) quantification and the method is discussed. The dialysis was optimized with Cr(VI) standard solutions for quantification. The efficacy of the procedure for the removal of anionic dyes and detection of Cr(VI) was checked with a Cr(VI) spiked synthetic preparation containing a water-soluble dye and trivalent chromium. Soluble Cr(VI) extracted with organic dyes from environmental samples was analyzed. The method has a detection limit of 5 microg/l, recovery rate of 100% and analysis time less than 20 min. PMID- 12647831 TI - Determination of organic acids in tobacco by capillary isotachophoresis. AB - Nine organic acids and phosphate have been separated and quantified in tobacco by capillary isotachophoresis. Two operating systems for the separation were found: hydrochloric acid (10 mmol l(-1)) adjusted with beta-alanine to pH 2.9 plus 0.1% poly(vinylpyrrolidone) was used as the leading electrolyte and 10 mmol l(-1) nicotinic acid as the terminating electrolyte and hydrochloric acid (10 mmol l( 1)) including 5.5 mmol l(-1) 1,3-bis[tris(hydroxymethyl)methylamino]propane plus 0.1% poly(vinylpyrrolidone) was used as the leading electrolyte and 5 mmol l(-1) 2-morpholinoethanesulfonic acid as the terminating electrolyte. Linearity was observed from 0.008 to 0.100 mmol l(-1) with a coefficient of determination (r2) of 0.999. The separation of anions was achieved in less than 16 min. The minimal sample pretreatment and relatively low running cost make isotachophoresis a good alternative to existing methods. PMID- 12647832 TI - Continuing education. PMID- 12647834 TI - When will immunomodulation for the prevention of type 1 diabetes be a reality? PMID- 12647833 TI - Exercise in type 1 diabetes. PMID- 12647835 TI - Triple oral anti-diabetic therapy in type 2 diabetes. PMID- 12647836 TI - Differing aspects of insulin resistance in diabetes complications: the shape of things to come. RD Lawrence Lecture 2000. AB - The anticipated global epidemic of diabetes, largely as a consequence of increased rates of obesity, will particularly impact on people of South Asian and African Caribbean descent, two populations at elevated risk of insulin resistance. This article contrasts the consequent heightened risk of heart disease on the one hand in South Asians, and the paradoxical protection from heart disease in African Caribbeans on the other. Protection from the hypertriglyceridaemic effects of insulin resistance is likely to account for much of the African Caribbean paradox, although the mechanisms remain unclear. The growing evidence that insulin resistance is commonly observed in people with Type 1 diabetes, as well as those with Type 2 diabetes, and that features of insulin resistance may play a crucial role in the development of microvascular, as well as macrovascular complications, is also discussed. This indicates novel targets for the prevention and treatment of diabetes complications. PMID- 12647837 TI - The status of diabetes control in Asia--a cross-sectional survey of 24 317 patients with diabetes mellitus in 1998. AB - AIMS: To establish the status of diabetes control in Asia, the Diabcare-Asia 198 study collected data from 230 diabetes centres in Bangladesh, People's Republic of China, India, Indonesia, Malaysia, Philippines, Singapore, South Korea, Sri Lanka, Taiwan, Thailand and Vietnam from March to December 1998. METHODS: Data were obtained either by patient interview during the enrolment visit or by reviewing medical records for the most recent laboratory assessment and clinical examinations. Blood samples were also collected during patients'. visits for central assessments of HbA1c (normal range 4.7-6.4%). RESULTS: The mean of centrally measured HbA1c was 8.6 +/- 2.0% for 18 211 patients (82% of the analysis population). Of the patients with central HbA1c measurements, the majority (55%) had values exceeding 8%, indicative of poor glycaemic control. The prevalence of retinopathy, microalbuminuria and neuropathy was also higher in the group of patients with higher HbA1c. Based on the findings from central HbA1c measurements and reported local HbA1c assessments, it also appears that more patients with poor glycaemic control did not have access to glycated haemoglobin measurements. Mean HbA1c of thediabetic populations in Bangladesh, Indonesia, Korea, Malaysia and Taiwan were significantly lower (all P = 0.0001, except P = 0.0007 for Malaysia), while that of China, India, Philippines and Vietnam was significantly higher (all P = 0.0001) than the grand mean. CONCLUSIONS: In our study population of the Asian diabetes patients treated at diabetes centres, more than half were not well controlled. The prevalence of diabetic microvascular complications was higher in the group of patients with higher HbA1c. Further therapeutic actions to improve glycaemic control are required to prevent chronic diabetic complications. PMID- 12647838 TI - Pertussis infection in childhood and subsequent type 1 diabetes mellitus. AB - AIMS: Pertussis has been implicated but not proven as a risk for Type 1 diabetes mellitus (DM). Previous studies have investigated paediatric, but not adult-onset Type 1 DM. We investigated association of pertussis exposures and Type 1 DM with follow-up into adulthood. METHODS: Longitudinal analysis of 16 820 members (100 with Type 1 DM) of two nationally representative British birth cohorts (the 1970 British Cohort Study (BCS70) and the National Child Development Study (NCDS)) followed from birth to ages 30 years (BCS70) and 42 years (NCDS). Cox regression analysis with age of onset for Type 1 DM as the dependent variable investigated relationships with pertussis infection and immunization, modelled as time dependent co-variates. Simultaneous adjustment was made for Wild measles, mumps and chickenpox infections; tetanus and smallpox immunizations; sex, parental social class and cohort. The potential confounding factors were modelled as fixed co-variates. RESULTS: Cox regression analysis produced adjusted odds ratios (ORs) (with 95% confidence intervals (CIs)) of 2.21 (1.35-3.59) and 0.73 (0.49-1.05) for Type 1 DM (with onset at any age) associated with pertussis infections and immunization (trend over number of vaccinations), respectively. Adjusted ORs from Cox regression for Type 1 DM with onset after age 10 years are 2.59 (1.56-4.30) for pertussis infection and 0.63 (0.42-0.94) for pertussis immunization. None of the other infections or immunizations are notably associated with Type 1 DM. CONCLUSIONS: Some pertussis infections may be a risk for Type 1 DM and immunization may confer protection. Further research should consider delayed Type 1 DM following pertussis exposures. PMID- 12647839 TI - Heritability of features of the insulin resistance syndrome in a community-based study of healthy families. AB - AIMS: To investigate genetic and environmental influences on anthropometric, metabolic and fibrinolytic traits of the insulin resistance syndrome (IRS) in a population not characterized by a high degree of insulin resistance. METHODS: We recruited 537 adults from 89 randomly ascertained healthy families of white north European origin from the general population. We used maximum likelihood analysis to estimate the heritabilities and effects of environmental covariates on traits of the IRS in these families. RESULTS: Adjusted for age, sex and body mass index, the traits showed considerable heritability. For waist-hip ratio, heritability was 15%. The heritabilities of fasting glucose, insulin and estimated insulin resistance were 20%, 23% and 23%, respectively. Heritabilities were 20%, 24% and 43% for triglycerides, LDL-cholesterol and HDL-cholesterol, respectively. For PAI 1 Ag and t-PA Ag they were 20% and 26%. Covariates explained 20-25% of the variance of lipids and insulin resistance and 35-36% of fibrinolytic factors. Childhood household influences significantly affected variance for waist-hip ratio (4%), fasting insulin (11%) and estimated insulin resistance (12%). CONCLUSIONS: These family data demonstrate significant genetic influence on anthropometric, fibrinolytic and glucose-related traits of the IRS in a healthy white North European population. PMID- 12647840 TI - Interleukin-6 polymorphism (-634C/G) in the promotor region and the progression of diabetic nephropathy in type 2 diabetes. AB - AIMS: Interleukin-6 (IL-6) is a multifunctional cytokine produced by many different cell types, including glomerular mesangial cells. Recently, a novel C/G polymorphism at position -634 in the promotor region of the IL-6 gene has been reported. The aim of this study was to investigate whether the -634C/G polymorphism is associated with an increased risk for progression to diabetic nephropathy as well as elevated levels of IL-6 secretion by peripheral blood mononuclear cells. METHODS: The frequency of the -634C/G polymorphism was determined in Japanese patients with Type 2 diabetes and either normoalbuminuria (n = 162), microalbuminuria (n = 138), or macroalbuminuria (n = 154) by polymerase chain reaction-restriction fragment length polymorphism analysis. The level of IL-6 secretion in relation to genotype was assessed in lipopolysaccharide or advanced glycation end products-stimulated IL-6 secretion by peripheral mononuclear cells. RESULTS: The frequency of the -634G/G genotype and -634*G allele was significantly increased in the patients with macroalbuminuria compared with patients with normoalbuminuria (genotype: chi2 = 6.787, Pc = 0.0368; allele: chi2 = 9.080, Pc = 0.0104). Stepwise multiple regression analysis in these patients showed that hypertension (F = 40.48) and IL 6-634 gene polymorphism (F = 5.48) were the relevant variables for the progression of Type 2 diabetic nephropathy. Analysis of the IL-6 secretion data revealed that individuals carrying the -634*G allele had a higher IL-6 secretion capacity than those without the *G allele (P < 0.05). CONCLUSIONS: These results suggest that the IL-6-634C/G polymorphism may be a possible genetic susceptibility factor for the progression of diabetic nephropathy. PMID- 12647841 TI - Preventing glycaemic excursions in diabetic patients requiring percutaneous endoscopic gastrostomy (PEG) feeding after a stroke. AB - AIMS AND METHODS: Enteral feeding for diabetic patients with a stroke is often associated with hyperglycaemia and/or hypoglycaemia, which can adversely influence neurological recovery. We have developed a structured enteral feeding programme aimed at establishing 'normal' feeding patterns and avoiding marked glycaemic excursions. RESULTS: Of 332 consecutive patients admitted to an acute stroke unit, 20 of 41 diabetic patients required PEG feeding. Over the initial 24 48 h, patients had 22- h feeds with continuous intravenous infusion of soluble insulin. Thereafter, they were established on three feeds per day. Soluble insulin was given prior to each feed with isophane insulin at 2200 h. Average duration of feed was 13 +/- 8 days with an achieved glucose level of 8.7 +/- 2.5 (mean +/- SD) mmol/l associated with 0.8 episodes of biochemical hypoglycaemia (< 3 mmol/l) each week. CONCLUSIONS: A structured enteral feeding programme for PEG fed diabetic patients can improve the quality of care with avoidance of marked glycaemic excursions. PMID- 12647842 TI - Ultrasonography of plantar soft tissues thickness in young people with diabetes. AB - AIMS: The aims of this study were to examine soft tissue changes in the skin and plantar aponeurosis of young people with Type 1 diabetes mellitus (T1DM) and to evaluate any relationship between any soft tissue changes, arch length, limited joint mobility (LJM) and plantar pressure. METHODS: The thickness of the skin on the plantar surface of the foot and plantar aponeurosis were examined using ultrasound in 216 young people with diabetes and 57 controls. Foot length, arch length, joint mobility, peak pressure and pressure time integrals were evaluated. RESULTS: Skin was not significantly thicker in the diabetic subjects. The plantar aponeurosis was significantly thicker in the diabetic subjects and was associated with foot size, male gender and subtalar joint (ST) LJM (P < 0.01). Males were nearly three times more likely to have thickened plantar aponeurosis. CONCLUSION: Soft tissue thickening in young people with T1DM affects the deeper structures on the plantar surface of the foot rather than the skin. Thickening of the plantar aponeurosis was associated with LJM at the ST joint and male gender, but was not associated with plantar pressure or arch height changes. Plantar aponeurosis thickening does not appear to alter foot mechanics in young people with T1DM. PMID- 12647843 TI - Prevalence of diabetic eye disease in patients entering a systematic primary care based eye screening programme. AB - AIMS: Large-scale, baseline prevalence measurements in a population at the institution of systematic retinal screening are currently unavailable. We report the prevalence of all grades of retinopathy at entry into a systematic primary care-based diabetic eye screening programme. METHODS: Primary care-based photographic screening utilizing mydriasis and three-field non-stereoscopic photography for all patients with diabetes (except those under continuing care of an ophthalmologist) in Liverpool. Sight-threatening diabetic eye disease (STED) was defined as any of: moderate preproliferative retinopathy or worse, circinate maculopathy or exudates within one disc diameter of the centre of fovea. RESULTS: Type 1 diabetes mellitus (DM) (n = 831): baseline prevalence (95% confidence interval (CI)) of any retinopathy, proliferative diabetic retinopathy (PDR) and STED was 45.7% (42.3-49.1), 3.7% (2.4-5.0) and 16.4% (13.9-18.9), respectively. Presence of STED was associated with increased disease duration (odds ratio (OR) 1.09 per year; P < 0.0001) and higher in men (OR 2.15; P = 0.001). Type 2 DM (n = 7231): baseline prevalence (95% CI) of any retinopathy, PDR and STED was 25.3% (24.3-26.3), 0.5% (0.3-0.7) and 6.0% (5.5-6.5), respectively. Presence of STED was associated with longer time since diagnosis of DM (OR 1.03; P < 0.0001) and insulin use (OR 2.46; P < 0.0001). CONCLUSION: This study provides baseline information for health providers on prevalence of all grades of retinopathy and STED in a large population at the establishment of systematic screening. Baseline prevalence of STED was high and highest in patients with a longer disease duration in both Type 1 and Type 2 DM. PMID- 12647844 TI - Post-mortem findings in familial partial lipodystrophy, Dunnigan variety. AB - AIMS: Familial partial lipodystrophy, Dunnigan variety (FPLD), is an autosomal dominant disorder due to missense mutations in the lamin A/C gene and is characterized by gradual loss of subcutaneous fat from the extremities and trunk, fat accumulation in the head, neck and intra-abdominal areas, insulin resistance and its metabolic complications. We studied autopsy findings in two patients with FPLD to determine fat distribution and organ involvement. RESULTS: Patient 1, a 66-year-old woman with the R482Q mutation, had diabetes mellitus, dyslipidaemia, and coronary artery disease and died suddenly. Autopsy confirmed the typical body fat distribution and further revealed excess fat deposition in the subpectoral regions extending to the axillae, in the axillary lymph nodes and in the retroperitoneum. Atherosclerotic vascular disease including old infarcts of the myocardium, temporal lobe and kidneys were noted. Severe amyloidosis of the pancreatic islets and grouped muscle atrophy of the quadriceps and diaphragmatic muscles were present. Patient 2, a 29-year-old woman belonging to a pedigree with the R62G mutation, died of hyperlipidaemia-induced acute pancreatitis. Autopsy of patient 2 revealed extensive pancreatitis, hepatic steatosis and polycystic ovaries. CONCLUSIONS: Our study confirms typical body fat distribution and describes new sites of excess fat deposition. Our data show predisposition to atherosclerosis and polycystic ovaries and suggest that pancreatic amyloidosis may underlie development of hyperglycaemia in FPLD patients. PMID- 12647846 TI - Microbiological profile of infected diabetic foot ulcers. PMID- 12647845 TI - Efficacy of Humalog injections before an afternoon meal and their acceptance by children and adolescents with type 1 diabetes. AB - AIMS: To evaluate the acceptability and efficacy of an injection of insulin lispro, before an afternoon meal. METHODS: The subjects, 43 patients with Type 1 diabetes, 16 boys and 27 girls, aged 12.4 +/- 2.4 years, were randomly assigned to the treatment (n = 20) or the untreated control group (n = 23). The treatment was an injection of insulin lispro immediately before the afternoon meal. The control group had no injection. The treatment and the control group consumed identical types of meals for 2 months. The mean before-dinner blood glucose was measured during the last 2 weeks of the study. RESULTS: Injection of insulin lispro resulted in a significant reduction in the before-dinner blood glucose compared with the untreated control group (10.4 +/- 3.8 mmol/l vs. 14.7 +/- 3.9 mmol/l, respectively). The number of days on which the blood glucose was > 10 mmol/l was reduced by half in the insulin lispro group. The difference in HbA1c between baseline and endpoint differed slightly but significantly between the two groups, in boys. Treated patients ate the meal less frequently (11.4 +/- 3.0 times per 15 days) than the control patients (14.4 +/- 0.6 times per 15 days) and injected themselves with insulin 8.9 +/- 3.6 times per 15 days. The HbA1c increased significantly with the number of meals taken without injection. There was no statistically significant difference in the frequency of hypoglycaemia or changes in weight between the two groups. CONCLUSIONS: We conclude that an injection of insulin lispro before the afternoon meal can effectively lower the before-dinner blood glucose, and in boys also lowers the HbA1c. Patients were satisfied with the lower blood glucose before dinner, and did not find the insulin lispro injection difficult. However, compliance with the protocol procedures decreased during a subsequent 6-month period. PMID- 12647847 TI - Continuous Internet-based activity monitoring provides a surrogate marker for nocturnal polyuria in persons with diabetes mellitus at high risk for foot ulceration. PMID- 12647849 TI - (S)-(+)-linalool, a mate attractant pheromone component in the bee Colletes cunicularius. AB - Enantiomerically pure (S)-(+)-linalool was the main constituent in the extracts of the cephalic secretions of virgin females, mated females, freshly emerged males, and patrolling males of the solitary bee Colletes cunicularius. After copulation, the content of (S)-(+)-linalool emitted by the female was strongly reduced. Electrophysiological experiments revealed that both enantiomers of linalool elicited responses from the antennae of the males. Field tests using the pure enantiomers and the racemate of linalool showed that the number of male bees attracted was highest for (S)-(+)-linalool. The search flight activity in the mating flight area increased dramatically when patrolling males were presented with (S)-(+)-linalool vs (R)-(-)-linalool. Taken together, these data indicate a mate attractant pheromone function of (S)-(+)-linalool. PMID- 12647850 TI - Sex pheromone of the scarab beetle Phyllophaga elenans and some intriguing minor components. AB - Three amino acid-derived compounds were identified in extracts from the pheromone glands of the scarab beetle Phyllophaga elenans, i.e., L-isoleucine methyl ester (LIME), N-formyl L-isoleucine methyl ester (For-LIME), and N-acetyl L-isoleucine methyl ester (Ac-LIME). The compounds were characterized from their spectral data (MS and IR), confirmed by synthesis, and their absolute configurations were assigned by gas chromatography with a chiral phase column. The amount of LIME in calling females was ca. 2 microg/gland, whereas only traces of For-LIME and Ac LIME (0.005% of LIME) were detected in fresh extracts. Regardless of the storage temperature, the amounts of the minor constituents in the extracts increased overtime. Field tests showed that traps baited with For-LIME captured more beetles than control traps. Ac-LIME per se was not attractive, and it did not increase trap catches when combined with For-LIME. Traps baited with LIME caught ca. 150 beetles/trap/day, but catches did not increase with the addition of For LIME and/or Ac-LIME in binary or tertiary blends. PMID- 12647848 TI - Alternative medicine recommendations to patients with type 2 diabetes visiting health product shops and pharmacies. PMID- 12647851 TI - Trapping of Phyllophaga elenans with a female-produced pheromone. AB - Attraction of Phyllophaga elenas to vaned bucket traps baited with the recently identified female-produced pheromone, L-isoleucine methyl ester (LIME), is efficient. Pheromone-baited vaned buckets with water to retain insects were more effective than buckets without vanes or plastic containers with the sides cut out. Pheromone-baited vaned bucket traps from which water was omitted required the addition of a funnel below the vanes to retain insects. Normally used light traps were about 10 times more effective than pheromone-baited vane bucket traps in capturing P. elenans. Over 95% of P. elenans were captured between 6:00 and 9:00 PM. The male-female ratio was approximately 3-4:1 in both light and pheromone traps, and the ratio was relatively unchanged throughout the capture period. Most P. elenans were captured in the treed areas surrounding sugarcane fields. More P. elenans were captured in treed borders than in grassy borders of sugarcane fields. The effective radius of the pheromone-baited vaned bucket trap is between 5 and 15 m. PMID- 12647852 TI - Behavioral and electrophysiological evidence for volatile sex pheromones in Parcoblatta wood cockroaches. AB - Species within the cockroach genus Parcoblatta are sexually dimorphic for wing length; females have reduced wings and are flightless, while males have long wings that are used in flight. We predicted that Parcoblatta females would release a volatile sex pheromone to attract the more mobile males. Nymphs of the broad wood cockroach, P. lata, and the Caudell's wood cockroach, P. caudelli, were collected in forested areas in North Carolina, USA, and reared in the laboratory for observations of sexual behavior and for pheromone analysis. After several days of sexual maturation, virgin females of both species exhibited distinct calling behaviors. In females of P. lata, calling commenced 6 days after adult emergence. Under a light-dark photoperiod regime, calling behavior in both species was restricted to the scotophase. Calling consisted of a repeated pattern of raising and lowering the abdomen with occasional exposure of the genital vestibulum. To test whether calling behavior is associated with the release of pheromone, volatiles from calling and noncalling females were collected on Super Q and tested by electroantennogram (EAG) and behavioral assays. Volatile collections from calling females elicited higher male-specific EAG responses than collections from noncalling females of the same physiological stage. In an olfactometer choice test (Y-tube), males preferred volatiles from calling females over those from noncalling females. To determine the anatomical source of the pheromone, solvent extracts of various body parts were analyzed by EAG. The first through seventh tergites were the only body parts that elicited male-specific EAG responses in both species. In P. lata, the activity of the extract increased from 1- to 7-day-old females, but was lower in mated than in virgin females of the same age. The putative pheromone gland appears to consist of numerous class-3 secretory units, each composed of a secretory cell connected to a cuticular pore via a tubular duct. We conclude that female P. lata and P. caudelli produce ex specific volatile pheromones that are emitted during calling behavior. PMID- 12647853 TI - Female sex pheromone-mediated effects on behavior and consequences of male competition in the shore crab (Carcinus maenas). AB - Exposure to receptive female pheromone elicits guarding behavior in shore crab males (Carcinus maenas), but little is known about the effects of sex pheromone on male competition or if the female plays an active role in mate choice. This study examined whether female pheromone enhanced agonistic behavior between males and what effects visual and chemical cues had on the rules and costs of such contests. We also investigated whether females exhibit a preference for males in terms of size. Under laboratory conditions, solitary male shore crabs engaged males who already had possession of a female. The visual and chemical presence of a receptive female had an impact on contest rules and costs. Fights were costly in terms of duration and of sustaining injury with either one or both crabs incurring injury in 40% of fights. To investigate the metabolic consequences of fighting over a perceived sexual resource (chemical cue only), fights were staged between pairs of size-matched males in the presence of water containing the female sex pheromone, water in which males had been kept, and untreated seawater. The duration and intensity of contests were greater when staged in the presence of the female pheromone compared with the two other treatments. Crabs that fought in the presence of female sex pheromone also had a greater accumulation of L lactate and a reduction of glycogen stores. Fights were less costly in terms of injury with a single chemical cue compared with enhanced costs with a multiple sexual cue. The importance of female choice was determined by presenting postmolt females with different sized males. Males were kept in a fixed position, and the majority of females approached and performed courtship behavior to the largest males, demonstrating that females may be selective in terms of size. PMID- 12647854 TI - Quantitative analysis of the effects of ultrasound from an odor sprayer on moth flight behavior. AB - A piezoelectric sprayer was recently developed for precision release of odor stimuli in olfactory research. The device replaces conventional dispensers used to release semiochemicals in studies of moth flight toward sources. However, the device generates high-frequency sounds in the range that some moths can hear. Ultrasound from the standard set-up sprayer had a considerable impact on flight behavior of the silver Y moth, Autographa gamma, tested in a flight tunnel. It was affected at all behavioral stages when the dispenser was driven at 120 kHz. Only 5% of the moths reached the source when exposed to 120-kHz sound from the dispenser compared to 65% in the control group without sound. The proportion taking flight was also reduced. Hearing threshold curves obtained electrophysiologically revealed that moths were sensitive to the frequency range at which the sprayer was operated and that sound intensity from the sprayer was up to 40 dB above the moths' electrophysiological hearing threshold. The audiogram for A. gamma was similar to audiograms obtained for other noctuids. Hearing sensitivity was highest at around 15 kHz, where the threshold was 35 dB SPL (sound pressure level). The threshold increased with frequency up to 94 dB SPL at 160 kHz. We improved the sprayer to operate at 300 kHz, which is beyond the hearing ability of most insects with ears. At this high frequency, the moths' sensitivity to ulrasound is reduced considerably, and we did not observe any effect on flight behavior compared to a control group without sound. Accordingly, this new piezoelectric sprayer can be used with ultrasound-sensitive insects and insensitive insects alike. PMID- 12647855 TI - Differential metabolism of 1,8-cineole in insects. AB - In order to compare the metabolism of 1,8-cineole in the pyrgo beetle, Paropsisterna tigrina, three other herbivorous insect species, Faex nigroconspersa, Chrysophtharta bimaculata, and Oxyops vitiosa, were fed 1,8 cineole leaf diets. F. nigroconspersa adults excreted predominantly 9-hydroxy-1,8 cineole (36.2% of the volatile constituents) with some 2alpha-hydroxy-1,8-cineole (11.4%). In contrast, larvae excreted predominantly 2alpha-hydroxy-1,8-cineole (27.4%) and smaller proportions of 9-hydroxy-1,8-cineole (5.2%) and 3alpha hydroxy-1,8-cineole (4.3%). C. bimaculata adults excreted predominantly 3alpha hydroxy-1,8-cineole (16.5%). Oxyops vitiosa adults, on a lower 1,8-cineole diet, excreted predominantly 2alpha,9-dihydroxy-1,8-cineole (4.2%) and 2alpha-hydroxy 1,8-cineole (3.5%), with smaller proportions of 3alpha-hydroxy-1,8-cineole (1.1%) and 9-hydroxy-1,8-cineole (0.5%). This is the first reported occurrence of a dihydroxycineole as an insect metabolite. Gas chromatographic and mass spectral data for hydroxycineoles are recorded and interspecific metabolite variation discussed. PMID- 12647856 TI - Comparative survey of abdominal gland secretions of the ant subfamily Ponerinae. AB - The chemical contents of three abdominal glands were investigated in representative species of the ponerine ants. The Dufour glands of 14 species show a wide variety of contents. In Mystrium camillae and Proceratium itoi, no volatile substances were found in either the Dufour or venom glands. In Ectatomma sp., Diacamma ceylonense, Diacamma indicum, Pachycondyla obscuricornis, and Pachycondyla striata, volatile chemicals were found in the venom glands as well as in the Dufour glands. Platythyrea punctata was examined, but unusually it does not have a Dufour gland and its venom gland contained no volatile substances. Epoxides were found in ants for the first time in the Dufour glands of Amblyopone reclinata. Venom glands of Pachycondyla tarsata were also found to contain volatile material, including bitter-tasting cyclic dipeptides. In all, 16 species have been added to the list of those examined. All of the 27 known analyses of Dufour glands, 21 analyses of venom glands, and 4 of pygidial glands of workers of ponerine ant species have been brought together in order to seek some pattern in the type of glandular contents. Although the great majority of species produce hydrocarbons in their worker Dufour glands, and some have terpenes, there is no observable pattern for this gland on a tribe or genus level. Volatile compounds have been found in the venom glands of some species of the tribe Ponerini only. The information on pygidial glands is still too fragmentary for any conclusions. PMID- 12647857 TI - Chemical cues from Murgantia histrionica eliciting host location and recognition in the egg parasitoid Trissolcus brochymenae. AB - Host location and recognition by the egg parasitoid Trissolcus brochymenae were analyzed in terms of response to kairomones from several stages of its host, Murgantia histrionica. In a Y-tube olfactometer, parasitoid females responded by increasing residence time and/or reducing linear speed to chemical cues from gravid females, virgin females and males, fifth and third instars, and eggs. In an open arena, T. brochymenae females also responded to patches contaminated by chemicals from the host in the same stages, sexes, and/or physiological conditions as those tested in the olfactometer. The parasitoid displayed arrestment behavior, increased residence time, changed walking pattern, and intense substrate examination. When host egg clusters or glass dummies with a chemical egg extract were placed on the host-contaminated open arena, these elicited an orientation response in the parasitoid. In addition, the chemical egg extract without dummies elicited the same response, whereas dummies without extract did not influence parasitoid behavior. In a closed arena, the parasitoid females recognized and attempted to probe glass beads treated with chemical extracts of host eggs. There were no significant differences compared with their response to the host eggs, and they did not respond to untreated beads. Host recognition was elicited by chemicals from the follicular secretion used by the host to glue the eggs on the substrate. These results are discussed in relation to the level of the host selection sequence influenced by these cues. PMID- 12647858 TI - Specific foraging kairomones used by a generalist parasitoid. AB - In general, it is assumed that generalist natural enemies do not innately use specific cues for the location of their host or prey species. This hypothesis was tested using naive females of the generalist parasitoid Lariophagus distinguendus Forster and two of its hosts, larvae of the lesser grain borer Rhyzopertha dominica (F.) and of the granary weevil Sitophilus granarius L., feeding in wheat grains. In a four-chamber olfactometer, female parasitoids were attracted to volatiles emanating from the feces of both host species. Chemical analysis of the volatiles from the feces of R. dominica revealed the presence of dominicalure 1 and 2, the species specific aggregation pheromones of R. dominica. The main compounds in the volatiles from feces of S. granarius were identified as chemicals related to mites that are associated with hosts of L. distinguendus. Because these mites are not specific for S. granarius but also co-occur with other hosts, the mite chemicals have to be considered as general cues. In bioassays, synthetic dominicalure was attractive to naive L. distinguendus, explaining the attraction of feces volatiles from R. dominica. Synthetic mite chemicals and sitophilate, the aggregation pheromone of S. granarius, had no effect on naive parasitoids. It remains to be determined which innate chemical cues from feces of S. granarius are used by L. distinguendus. In contrast to our initial hypothesis, the generalist L. distinguendus is innately using specific cues for foraging. Two ideas are provided to explain this result. PMID- 12647859 TI - Induction of volatile emissions in maize by different larval instars of Spodoptera littoralis. AB - Maize plants under attack by caterpillars emit a specific blend of volatiles that is highly attractive to parasitic wasps. The release of these signals is induced by elicitors in the caterpillar regurgitant. Studies suggest that plants respond differently to different herbivore species and even to different herbivore stages, thus providing parasitoids and predators with specific signals. We tested if this is the case for different larval instars of the noctuid moth Spodoptera littoralis when they feed on maize plants. Cut maize plants were incubated in diluted regurgitant from second, third, or fifth instar caterpillars. There were no differences in total amount released after these treatments, but there were small differences in the release of the minor compounds phenethyl acetate and alpha-humulene. Regurgitant of all three instars contained the elicitor volicitin. To test the effect of actual feeding by the larvae, potted plants were infested with caterpillars of one of the three instars, and volatiles were collected the following day. The intensity of the emissions was correlated with the number of larvae feeding on a plant, and with the amount of damage inflicted, but was independent of the instar that caused the damage. We also used artificial damage to mimic the manner of feeding of each instar to test the importance of physical aspects of damages for the odor emission. The emission was highly variable, but no differences were found among the different types of damage. In olfactometer tests, Microplitis rufiventris, a parasitoid that can only successfully parasitize second and early third instar S. littoralis, did not differentiate among the odors of maize plants attacked by different instar larvae. The odor analyses as well as the parasitoid's responses indicate that maize odors induced by S. littoralis provide parasitoids with poor information on the larval developmental stage. We discuss the results in the context of variability and lack of specificity in odorous plant signals. PMID- 12647860 TI - Salicylic acid mediates resistance in the willow Salix viminalis against the gall midge Dasineura marginemtorquens. AB - Resistant willow (Salix viminalis L.) genotypes react with a rapid hypersensitive response (HR) when attacked by the gall midge Dasineura marginemtorquens Bremi. In general, infected plant genotypes or species that react hypersensitively to pathogens accumulate salicylic acid (SA) locally and systemically. Thus, pathogen induced HRs are strongly associated with accumulation of SA. In this study, we tested the hypothesis that SA mediates resistance in S. viminalis against D. marginemtorquens. By means of HPLC, we found accumulation of SA in a resistant S. viminalis genotype, but not in a susceptible genotype one to two days after D. marginemtorquens egg hatch. To test whether this correlation between accumulation of SA and resistance is causal, we treated a normally susceptible S. viminalis genotype exogenously with SA and found a decreased larval survival frequency on treated shoots. Thus, the hypothesis that SA mediates resistance in the S. viminalis/D. marginemtorquens system was strongly supported. Interestingly, great similarity seems to exist between the biochemical signaling associated with pathogen-induced HRs and this gall-inducer induced HR. PMID- 12647861 TI - Allelochemical effects of eudesmane and eremophilane sesquiterpenes on Tribolium castaneum larvae. AB - Eight eudesmane and eremophilane sesquiterpenes administered to Tribolium castaneum larvae caused different allelochemical effects. Topical application of 3-oxo-gamma-costic acid produced the greatest lengthening in the duration of the pupal stage. Morphological deformities were found, specifically when ilicic, costic, and gamma-costic acids and costic aldehyde were used. Ilicic acid exhibited the major toxicity 72 hr following topical application. All compounds were significantly toxic at the end point of the experiment (60 days). Treated surface toxicity was lower than when topical assays were carried out. Responses to tessaric acid in choice bioassays had the highest attractive effect. Maximum repellency was caused by the 3-oxo-gamma-costic acid. However, experimental series carried out using gamma-costic acid, eremophilan-1(10),2,11(13)-trien-12 oic acid, costic aldehyde, and ilicic aldehyde showed no clear response. PMID- 12647862 TI - Dietary influences on terpenoids sequestered by the biological control agent Oxyops vitiosa: effect of plant volatiles from different Melaleuca quinquenervia chemotypes and laboratory host species. AB - The weevil Oxyops vitiosa is an Australian species imported to Florida, USA, for the biological control of the invasive species Melaleuca quinquenervia. The larvae of this species feed on the leaves of their host and produce a shiny orange secretion that covers their integument. Previous results indicated that a major component of this secretion, viridiflorol, is sequestered from the host plant and repels a generalist predator, the red imported fire ant, Solenopsis invicta. When the larvae fed on a different chemotype of M. quinquenervia, which lacked viridiflorol but was rich in a different sesquiterpene, (E)-nerolidol, similar protection occurred. Solvent washes of these larvae indicated that (E) nerolidol was sequestered from M. quinquenervia leaves and repelled S. invicta workers when applied to dog food baits at physiological concentrations (17.5, 35.0, and 52.5 microg/mg). Additionally, beta-caryophyllene also repelled S. invicta workers when applied to dog food baits at concentrations that approximated those in the O. vitiosa larval secretions (3.5 and 35 microg/mg). When the O. vitiosa larvae were fed leaves from laboratory hosts (nonfield hosts), similar repellent activity was found. This activity was traced to several of the same compounds (e.g., 1,8-cineole, viridiflorol) found to be active in their field host M. quinquenervia. These weevil larvae are opportunistic, sequestering the primary terpenoids in their host leaves that confer antipredator activity. PMID- 12647863 TI - Cynipid gall-wasp communities correlate with oak chemistry. AB - Host-plant association data, gathered from field surveys conducted throughout Florida and from the literature, were used to identify the specificity of cynipid gall inducers to one or more of six Quercus species that occur at Archbold Biological Station, Lake Placid, Florida, USA, including the red oaks Q. laevis, Q. myrtifolia, and Q. inopina, and the white oaks Q. chapmanii, Q. geminata, and Q. minima. Quercus myrtifolia had the highest cynipid richness and diversity (37 cynipid species, Shannon H' = 3.61, Simpson's D = 0.97), followed by Q. chapmanii, Q. laevis, Q. inopina, Q. geminata, and finally Q. minima (10 species, H' = 2.30, D = 0.90). All cynipid species showed strong fidelity to a particular host plant or a restricted set of host plants. An ordination of gall-wasp host associations indicated that the cynipid communities of each oak species were distinct and specific to a given oak species. Leaf samples taken from each oak species were analyzed for condensed and hydrolyzable tannins, total phenolics, lignin, cellulose and hemicellulose, nitrogen, and carbon. All of these chemical traits, with the exception of carbon, differed by oak species, and the differences were strongly correlated with the axes of the cynipid-species ordination. These results suggest that gall-wasp occurrence is influenced by oak chemistry and imply that experimental studies of cynipid gall inducers that examine host-plant chemistry and female oviposition choice and larval performance will yield useful insights. PMID- 12647864 TI - Role of cucurbitacin C in resistance to spider mite (Tetranychus urticae) in cucumber (Cucumis sativus L.). AB - Cucurbitacins are bitter triterpenoid compounds that are toxic to most organisms and occur widely in wild and cultivated Cucurbitaceae. The only cucurbitacin identified in Cucumis sativus is cucurbitacin C. The bitter taste of cucumber has been correlated with resistance to the spider mite Tetranychus urticae, but a quantitative relationship has not been established. We determined the spider mite resistance and cucurbitacin C content in the dihaploid progeny derived from the F1 generation of a cross between a bitter, spider-mite-resistant cucumber line and a bitter-free, spider-mite-susceptible line. The ratio of the number of bitter to bitter-free dihaploids conformed to the expected 1:1 ratio, based on a monogenic segregation pattern. Genetic analysis ascribed 69% of the variance of the difference in spider mite survival rate to the bitterness locus. Within the group of bitter dihaploids, cucurbitacin C content was significantly correlated with spider mite resistance. Thus, a quantitative relationship between cucurbitacin C content and spider mite resistance could be established. PMID- 12647865 TI - Effect of smoke, charred wood, and nitrogenous compounds on seed germination of ten species from woodland in central-western Spain. AB - The effect of smoke, charred wood, and nitrogenous compounds on germination was tested on 10 species of the Cistaceae, Poaceae, Fabaceae, and Asteraceae, from fire-prone, shrubby woodlands in central-western Spain. Dry seeds were exposed to smoke, by watering with distilled water-charred wood suspensions, or NaNO2, KNO3, NH4Cl, and NH4NO3. Smoke enhanced germination in 9 of 10 of the species. In species of Poaceae, germination was stimulated by 20 min of smoke exposure. In Asteraceae and Fabaceae species, 10 min of smoke exposure was the most effective treatment for enhancing germination. Three species--Cistus ladanifer, Cistus crispus, and Cistus monspeliensis--had a positive response to 20 min of smoke exposure; germination of Cistus salviifolius L. was also enhanced after 10 min. The effect of charred wood was variable, with no consistent germination pattern within the families. Trifolium angustifolium and Retama sphaerocarpa showed no stimulation of germination under most of the charred wood concentrations. Similarly, germination of Senecio jacobea under the charred wood treatment did not surpass that of the control. NaNO2 promoted seed germination in Dactylis glomerata (10 mM), Cistus ladanifer (1, 10, and 25 mM), and Cistus crispus (1 and 10 mM). KNO3 enhanced germination in Dactylis glomerata (1 and 25 mM), Dittrichia viscosa (10 and 25 mM), C. ladanifer (1, 10, and 25 mM), Cistus crispus (1 and 25 mM), and C. salviifolius aud C. monspeliensis (25 mM). NH4Cl induced germination of Dactylis glomerata and Dittrichia viscosa (1 mM), and Cistus species germinated best in 25 mM of this salt. NH4NO3 induced germination only in Cistus species. Holcus lanatus had the highest level of germination regardless of treatment. PMID- 12647866 TI - Odor compound detection in male euglossine bees. AB - Male euglossine bees collect fragrances from various sources, which they store and use for as yet unknown purposes. They are attracted, often specifically, to single odor compounds and blends thereof. We used gas chromatography with electroantennographic detection (GC-EAD) and electroantennography (EAG) to investigate the response to 8 odor compounds by males of two euglossine species, Euglossa cybelia Moure and Eulaema polychroma (Mocsary). In E. cybelia, we recorded EAD reactions in response to 1,8-cineole, methyl benzoate, benzyl actetate, methyl salicylate, eugenol, and methyl cinnamate. E. polychroma responded to the same compounds in EAG experiments, while (1s)(-)alpha-pinene and beta-pinene failed to trigger EAD or EAG responses in the bees. Blends of two compounds triggered larger responses than single compounds in EAG experiments with E. polychroma, however, when alpha-pinene was added, reactions decreased. In the light of existing data on the bees' behavior towards these odor compounds, our work indicates that both peripheral and central nervous processes influence the attraction of euglossine bees to odors. PMID- 12647867 TI - Optical projection tomography as a new tool for studying embryo anatomy. AB - Optical projection tomography (OPT) is a new technique for three-dimensional (3D) imaging of small biological tissues. It is particularly useful for reconstructing vertebrate embryos and for examining the 3D anatomy of developing organs. The advantages of this technique over previous methods will be explained: in particular, its ability to image at a higher resolution than magnetic resonance imaging (MRI), while at the same time being able to image specimens much larger than those possible using confocal laser-scanning microscopy. Being an optical technique, OPT is also able to take advantage of the many coloured and fluorescent dyes which have been developed for tissue-specific or gene-specific staining. This becomes particularly important for the visualization of the 3D shapes of specific organs and tissues as it allows the computer to automatically determine the outline of the desired structure. PMID- 12647868 TI - Improved digital SEM of cancellous bone: scanning direction of detection, through focus for in-focus and sample orientation. AB - Three-dimensional (3D) study of cancellous bone tissue organization is necessary to understand how modelling and remodelling processes regulate bone structure and connectivity. It requires imaging methods that have both sufficient resolution power and width and depth of field. Since clinical imaging methods fall far short of the first requirement, we can only study prepared tissue in isolation from the body. Scanning electron microscopy (SEM) of macerated plane parallel slices is the most productive method, but we meet special technical problems in imaging porous bone because samples need to be relatively thick to maintain both continuity and context. Problems due to charging under the electron beam can be controlled by imaging with only high-energy backscattered electrons (BSE). This gives an important additional benefit that the direction of apparent illumination can be manipulated by positioning the detector, and multiple detector positions can be employed strategically to generate images in which colour is used to help in coding surface morphology. However, we next confront the difficulty of the limited depth of field. This can be improved by taking series of images, moving the sample along the electron optic axis, and combining these to generate a single extended-focus image. SEM imaging geometry gives a change in magnification with change of working distance, and it is shown that this must be corrected for each image of the through-focus sequence. Colour coding the lighting direction and increasing the depth of field are approaches that can be combined, and are well matched to the possibilities offered by communication by digital data projection. Finally, the latter means also offer another powerful technique for 3D representation through the display of through tilt image sequences. The novel routines considered here are generally applicable to all classes of microanatomical SEM sample. PMID- 12647869 TI - Immunocytochemical investigation of neurovascular relationships in human tooth pulp. AB - This study sought to explore the anatomical relationships between peptidergic nerves and blood vessels within human primary and permanent teeth. Extracted primary and permanent molars (n = 120) were split longitudinally, placed in Zamboni's fixative and the coronal pulps were processed for indirect immunofluorescence. Ten-micrometre-thick serial frozen pulp sections were triple labelled using combinations of the following antisera: (1) protein gene-product 9.5 (PGP 9.5), a general neuronal marker; (2) one of the neuropeptides, calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal polypeptide (VIP) or neuropeptide Y (NPY); and (iii) the lectin Ulex europeus, a label for vascular endothelium. The mid-coronal pulp region was examined, using fluorescence microscopy, to determine the proportion of blood vessels showing a positive innervation (recorded when PGP 9.5-labelled nerves appeared to intersect the vessel wall). In addition, the percentage of these vascular-related nerves expressing each of the above neuropeptides was recorded. Overall, 20% of pulpal blood vessels appeared to have a positive innervation. In the main these were thick-walled arterioles. Capillaries, venules and lymphatics were mostly devoid of an associated innervation. Ninety-two per cent of vascular-related nerves expressed CGRP, 87% expressed SP, 15% expressed VIP and 80% expressed NPY. There were no significant differences in overall innervation or peptide-related innervation between primary and permanent teeth (P < 0.05, ANOVA), indicating that pulpal blood flow is likely to be subject to similar neurological control mechanisms in both dentitions. PMID- 12647870 TI - NADH-O2 oxidoreductase activity and mRNA expression of complex I (51 kDa, ND1) in postnatal intrinsic muscle of rat tongue. AB - Complex I is one of the respiratory chain enzymes related to NADH dehydrogenase and is an encoded gene product derived from both nuclear and mitochondrial genomes. Transcription levels of ND1 (mitochondrial) and 51 kDa (nuclear) subunits of complex I in the postnatal development of the intrinsic muscle in rat tongues were determined by Northern blot analysis. Enzyme activity levels were determined by NADH staining with tetrazolum salt, and oxygen consumption of NADH O2 oxidoreductase activity using a Clark-type electrode. The detailed structure of the mitochondria was observed using electron microscopy. The cross-sectional area of the mitochondria gradually increased during postnatal development, and the cristae also became complex, despite the length of mitochondria in muscle fibre being constant. The mitochondria density increased from birth to 15 days of age, and declined slightly afterwards. This pattern of density resembled that of NADH-O2 oxidoreductase activity. The level of mRNA for ND1 through Northern blot analysis gradually increased from birth to 15 days of age and was highest at 21 days. For 51 kDa, the level was highest at 0 days and fell thereafter to a constant low. This suggests that the production of NADH dehydrogenase is limited by 51 kDa of Complex I derived from nuclear genomes rather than by the increase in mitochondria and composition of muscle fibre types due to changes in feeding behaviour. PMID- 12647871 TI - Microvascular assembly and cell invasion in chick mesonephros grafted onto chorioallantoic membrane. AB - Embryonic tissues, in common with other tissues, including tumours, tend to develop a substantial vasculature when transplanted onto the chorioallantoic membrane (CAM). Studies conducted to date have not examined in any detail the identity of vessels that supply these grafts, although it is known that the survival of transplanted tissues depends on their ability to connect with CAM vessels supplying oxygen and nutrients. We grafted the mesonephros, a challenging model for studies in vascular development, when it was fully developed (HH35). We used reciprocal chick-quail transplantations in order to study the arterial and venous connections and to analyse the cell invasion from the CAM to the organ, whose degeneration in normal conditions is rapid. The revascularization of the grafted mesonephros was produced by the formation of peripheral anastomoses between the graft and previous host vasculatures. The assembly of graft and CAM blood vessels occurred between relatively large arteries or veins, resulting in chimeric vessels of varying morphology depending on their arterial or venous status. Grafts showed an increased angiogenesis from their original vasculature, suggesting that the normal vascular degeneration of the mesonephros was partially inhibited. Three types of isolated host haemangioblast were identified in the mesonephros: migrating angioblast-like cells, indicating vasculogenesis, undifferentiated haematopoietic cells and macrophages, which might have been involved in the angiogenesis. Tomato lectin was found to bind activated macrophages in avian embryos. PMID- 12647873 TI - High-resolution imaging of normal anatomy, and neural and adrenal malformations in mouse embryos using magnetic resonance microscopy. AB - An efficient investigation of the effects of genetic or environmental manipulation on mouse development relies on the rapid and accurate screening of a substantial number of embryos for congenital malformations. Here we demonstrate that it is possible to examine normal organ development and identify malformations in mouse embryos by magnetic resonance microscopy in a substantially shorter time than by conventional histology. We imaged embryos in overnight runs of under 9 h, with an operator time of less than 1 h. In normal embryos we visualized the brain, spinal cord, ganglia, eyes, inner ear, pituitary, thyroid, thymus, trachea, bronchi, lungs, heart, kidneys, gonads, adrenals, oesophagus, stomach, intestines, spleen, liver and pancreas. Examination of the brain in embryos lacking the transcriptional coactivator Cited2 showed cerebellar and midbrain roof agenesis, in addition to exencephaly. In these embryos we were also able to detect agenesis of the adrenal gland. We confirmed all malformations by histological sectioning. Thus magnetic resonance microscopy can be used to rapidly identify developmental and organ malformations in mutant mouse embryos generated by transgenic techniques, in high-throughput mutagenesis screens, or in screens to identify teratogenic compounds and environmental factors contributing to developmental malformations. PMID- 12647872 TI - Constitutive expression of the low-affinity neurotrophin receptor and changes during axotomy-induced death of sensory neurones in the neonatal rat dorsal root ganglion. AB - Sensory neurones in the dorsal root ganglion (DRG) of the neonatal rat express the 75-kDa low-affinity neurotrophin receptor (p75NTR) and these neurones degenerate rapidly after axotomy. p75NTR belongs to the tumour necrosis factor superfamily, several members of which have a role in cell death and it is constitutively expressed within a subpopulation of DRG neurones. p75NTR has been implicated in mediating the degeneration of these neurones after axotomy. In this study, we characterize the expression of p75NTR in sensory neurones of the newborn rat DRG using immunohistochemistry. Furthermore, we investigate the change in constitutive expression pattern of p75NTR in these neurones following axotomy. In the C7 and C8 DRG of the newborn rat, p75NTR is expressed in approximately 70% of DRG neurones. Those expressing p75NTR can be classified into subpopulations with moderate or intense p75NTR expression, each present in approximately equal proportions. Whilst p75NTR expression is observed in neurones throughout the entire neuronal diameter range, a correlation exists between neuronal diameter and p75NTR expression intensity. We also found that the most vulnerable population following axotomy were those sensory neurones which constitutively express the highest levels of p75NTR, i.e. the large-diameter neurones. PMID- 12647875 TI - Radiofrequency energy: applications and basic science. AB - Interest in using radiofrequency energy devices in orthopedics has been growing recently. The basic science of this intervention is similar to that of laser energy application and involves the heating of tissue. Collagen responds characteristically to heat--it becomes denatured, and its configuration becomes disorganized. Thus, this application of heat can be used to shorten collagenous soft tissue. Radiofrequency energy can also be used at higher parameters for tissue ablation. Although early clinical trials have shown promising results for many applications, long-term study is needed. PMID- 12647874 TI - Wrong site surgery. PMID- 12647876 TI - Analysis of rotator cuff muscles in adult human cadaveric specimens. AB - Management of irreparable massive rotator cuff tears remains a challenging and controversial problem. Defining glenohumeral force relations may allow for the development of treatment strategies based on biomechanical principles. Five fresh frozen adult human cadaveric shoulder specimens were dissected to determine fiber length, mass, and lever arm of (a) the 3 bellies of the deltoid and (b) the rotator cuff muscles (supraspinatus, infraspinatus, teres minor, subscapularis). From these data, physiologic cross-sectional areas and moment relations were calculated. These relations provide evidence for a balanced axial force couple between the anterior and posterior rotator cuff. Demonstration of an axial force couple across the glenohumeral joint may have clinical significance for treatment of irreparable massive rotator cuff tears and may explain why many patients with full-thickness rotator cuff tears can regain acceptable shoulder function. PMID- 12647877 TI - Administering caudal anesthesia at completion of clubfoot surgery does not affect postoperative use of narcotics. AB - In the pediatric population, control of postoperative pain is a challenging and important issue. We conducted this retrospective study to determine whether single-dose caudal anesthesia administered after club-foot surgery helps to decrease postoperative use of narcotics. Fifty-one patients given an injection of caudal anesthesia (bupivacaine) at completion of clubfoot surgery were compared with 41 patients who did not receive a caudal block. Postoperative pain control was assessed by recording how much narcotic was used by each patient during time in the recovery room and during the first 8 hours after surgery. Results show that a single dose of caudal anesthesia administered at completion of clubfoot surgery is not associated with a statistically significant change in use of narcotics during either postoperative period. PMID- 12647878 TI - Compliance in using a pneumatic compression device after total knee arthroplasty. AB - A prospective study was performed to assess compliance with using a pneumatic foot compression device for venous thrombosis prophylaxis after total knee arthroplasty. One hundred patients received the PlexiPulse pneumatic compression device after unilateral total knee arthroplasty. Patients were observed postoperatively by the nurses on their floor and by 2 impartial research assistants. Total time the patient spent wearing the device (as observed) was termed total compliance. Actual compliance was calculated by subtracting unusable time from total compliance. A survey was also issued to both patients and nurses to evaluate the device regarding comfort, ease of use, and effectiveness. Overall, actual patient compliance was 90.1%. Patients responded favorably to the device in terms of comfort and ease of application and removal. Nurses rated the PlexiPulse device highly when asked to compare it with other pneumatic compression devices they had used before. PMID- 12647879 TI - Extensive lipoma causing suprascapular nerve entrapment. AB - Suprascapular nerve entrapment is a frequently misdiagnosed phenomenon. In this case report, we describe a benign lipomatous tumor causing suprascapular nerve entrapment in the spinoglenoid notch, and we describe the history, clinical examination, and radiographic evaluation. Benign lipomatous tumor compressing the suprascapular nerve is mentioned in the literature, but we believe this is the first case report. PMID- 12647880 TI - Solitary bilateral palmar masses as initial presentation of sarcoidosis. AB - Sarcoidosis, a systemic disease characterized by noncaseating granulomas, has multiple presentations in the hand. In this report, we present the case of a patient manifesting sarcoidosis initially as bilateral palmar nodules not involving bone. To our knowledge, similar cases have not been reported in the literature. PMID- 12647882 TI - In-hospital mortality after femoral neck fracture: do internal fixation and hemiarthroplasty differ? AB - In this article, we examine rates of in-hospital mortality of elderly patients with femoral neck fracture treated with internal fixation or hemiarthroplasty. Data were analyzed for 51,003 patients (> or = 65 years old) admitted with femoral neck fractures to New York state hospitals between 1985 and 1996. The primary outcome examined was in-hospital mortality. Associations between type of surgical procedure and outcome were assessed using a multiple logistic regression model, adjusting for patient age, sex, race, number of comorbidities, and residence in a nursing facility before hip fracture. Approximately 30% of the study group had undergone open or closed reduction and internal fixation; the other 70% had undergone hemiarthroplasty. Forty-six percent of the internal fixation group and 56% of the hemiarthroplasty group were 85 years old or older (P < .001). Median hospital stays were 13 days for the internal fixation group and 15 days for the hemiarthroplasty group (P < 001). In-hospital mortality was 5.1% overall, 3.9% for the internal fixation group, and 5.6% for the hemiarthroplasty group (P < .001). The association between type of procedure and mortality held after adjusting for patient age, sex, and number of comorbidities (odds ratio, 1.42; 95% confidence interval, 1.29-1.56; P < .001). After controlling for potential confounding variables, we found that elderly patients who had undergone hemiarthroplasty after femoral neck fracture were more likely to die during hospitalization than those who had undergone internal fixation. PMID- 12647881 TI - Primary bacterial pyomyositis associated with septic arthritis caused by Streptococcus pyogenes: a case report. AB - Pyomyositis is relatively rare in temperate climates. This is the first report of a case of calf pyomyositis associated with septic arthritis of the knee. This case illustrates that pyomyositis should be included in the differential diagnosis of a swollen, tender calf in an emergency setting. The other notable feature of this case is that the infective organism was not the usual Staphylococcus aureus but rather the relatively rarely involved Streptococcus pyogenes. PMID- 12647883 TI - Percutaneous toggle technique for manipulating displaced, unstable, and nonreducible proximal tibial fractures in children. AB - Reducing displaced and unstable tibial fractures of the proximal metaphysis can be challenging. Closed reduction with subsequent casting may fail to achieve acceptable anatomic alignment, potentially resulting in deformity or increasing the risk for compartment syndrome. A method is presented for aligning the opposing fragments by inserting a guide pin percutaneously to manipulate the proximal and distal segments in preparation for percutaneous Kirschner-wire stabilization. Using this percutaneous toggle technique lessens the chance that open reduction will be needed and thus avoids the inherent risks associated with open procedures. PMID- 12647884 TI - Intonation abilities of children with speech and language impairments. AB - Intonation has been little studied in children with speech and language impairments, although deficits in related aspects of prosody have been hypothesized to underlie specific language impairment. In this study a new intonation battery, the Profiling Elements of Prosodic Systems-Child version (PEPS-C), was administered to 18 children with speech and/or language impairments (LI). PEPS-C comprises 16 tasks (8 x 8, Input x Output) tapping phonetic and functional aspects of intonation in four areas: grammar, affect, interaction, and pragmatics. Scores were compared to a chronological age (CA) matched group of 28 children and a group of 18 children matched for language comprehension (LC). Measures of language comprehension, expressive language, nonverbal intelligence, and segmental phonology were also taken. The LI group did not score significantly below the LC group on any PEPS-C task. On 5 of 16 tasks, the LI group scored significantly lower than the CA group. In the LI group, there were just 2 significant correlations between a PEPS-C task and 1 of the nonprosodic measures. The results support the view that intonation is relatively discrete from other levels of speech and language while suggesting some specific areas of possible vulnerability: auditory memory for longer prosodic strings and the of prosody for pragmatic/interactional purposes. PMID- 12647885 TI - Inferring emotional reactions in social situations: differences in children with language impairment. AB - Anticipating and responding to a partner's emotional reactions are key components in the comprehension of daily social discourse. Kindergarten children with language impairment (LI) and age-matched controls (CA) were asked to label facial expressions depicting 1 of 4 emotions (happy, surprised, sad, mad) and to identify those expressions when given a verbal label. Children then chose among these facial expressions when asked to infer emotional reactions from stories (3 sentence scenarios) presented in 1 of 3 modalities: verbal, visual, and combined. Although all children were able to identify and label the facial expressions, children with LI had difficulty integrating emotion knowledge with event context in order to infer a character's feelings. When these inferencing errors occurred, children in the LI group were more likely to provide emotions of a different valence (e.g., substituting happy for mad) than were children in the CA group. Inferencing ability was related to language comprehension performance on a standardized test. The findings suggest that inferencing errors made by children with LI occur during the early stages of social processing and may contribute to social difficulties often experienced by this group of children. PMID- 12647886 TI - Auditory temporal processing in children with specific reading disability with and without attention deficit/hyperactivity disorder. AB - The auditory temporal deficit hypothesis predicts that children with specific reading disability (RD) will exhibit a deficit in the perception of auditory temporal cues in nonspeech stimuli. Tasks assessing perception of auditory temporal and nontemporal cues were administered to children with (a) RD without attention-deficit/hyperactivity disorder (RD/no-ADHD, n = 40), (b) ADHD alone (ADHD/no-RD, n = 33), (c) RD and ADHD (RD/ADHD, n = 36), and (d) no impairment (NI, n = 41). The presence of RD was associated with a specific deficit in detection of a tone onset time asynchrony, but no reduction in performance on other tasks assessing perception of temporal or nontemporal acoustic cues. The presence of ADHD was associated with a general reduction in performance across tasks. The pattern of results did not indicate a pervasive deficit in auditory temporal function in children with RD, but did suggest a possible sensitivity to backward masking in this group. Results also indicated that the comorbid presence of ADHD is a significant factor in the performance of children with RD on psychoacoustic tasks. PMID- 12647887 TI - Surface forms and grammatical functions: past tense and passive participle use by children with specific language impairment. AB - Children with specific language impairment (SLI) use past tense -ed in fewer obligatory contexts than younger normally developing children matched for mean length of utterance (MLU). In this study, the use of passive participle -ed (e.g., kissed in The frog got kissed by the kitty) as well as past tense -ed was examined in children with SLI, normally developing children matched for age (ND A), and normally developing children matched for MLU (ND-MLU). The children with SLI used both past tense -ed and passive participle -ed in fewer obligatory contexts than bath the ND-A and the ND-MLU children. Only the children with SLI had greater difficulty with past tense -ed than with passive participle -ed. The pattern of findings indicates that the surface properties of -ed cannot adequately account for the past tense -ed difficulty shown by the children with SLI. However, the fact that the children with SLI were less consistant than the ND-MLU children in using passive participle -ed suggests that either the surface properties of -ed are responsible for a portion of the difficulty or these children have a separate, non-tense-related deficit in the area of verb morphology. PMID- 12647888 TI - Word-learning by preschoolers with specific language impairment: what predicts success? AB - Thirty preschool children with specific language impairment (SLI) and 30 age matched controls with normal language (NL) participated in a study to compare group performance and to examine the relationship between fast mapping and word learning and between comprehension and production of new words. The groups performed similarly on the fast-mapping task. The NL group comprehended and produced significantly more words than the SLI group, and did so in fewer trials. Language test scores did not predict word-learning performance for either group. Some children with SLI may need to hear a new word twice as many times as their NL peers before comprehending it and may need twice as many opportunities to practice producing the word before using it independently. PMID- 12647889 TI - The perception of "sine-wave speech" by adults with developmental dyslexia. AB - Numerous studies have shown that, as a group, children or adults with developmental dyslexia perceive isolated syllables or words abnormally. Continuous speech containing reduced acoustic information also might prove perceptually difficult to such listeners. They might, however, exploit the intact syntactic and semantic features present in whole utterances, thereby compensating fully for impaired speech perception. "Sine-wave speech" sentences afford a test of these competing possibilities. The sentences contain only 4 frequency modulated sine waves, lacking many acoustic cues present in natural speech. Adults with and without dyslexia were asked to orally reproduce 9 sine-wave utterances, each occurring in 4 immediately successive trials. Participants with dyslexia reported fewer words than did control listeners. Practice, phonological contrasts, and word position affected both groups similarly. Comprehension of sine-wave sentences seems impaired in many, but not all, adults with dyslexia. A reduced auditory memory capacity may contribute to this deficit. PMID- 12647890 TI - Speaking clearly for children with learning disabilities: sentence perception in noise. AB - This study compared the speech-in-noise perception abilities of children with and without diagnosed learning disabilities (LDs) and investigated whether naturally produced clear speech yields perception benefits for these children. A group of children with LDs (n = 63) and a control group of children without LDs (n = 36) were presented with simple English sentences embedded in noise. Factors that varied within participants were speaking style (conversational vs. clear) and signal-to-noise ratio (-4 dB vs. -8 dB); talker (male vs. female) varied between participants. Results indicated that the group of children with LDs had poorer overall sentence-in-noise perception than the control group. Furthermore, both groups had poorer speech perception with decreasing signal-to-noise ratio; however the children with LDs were more adversely affected by a decreasing signal to-noise ratio than the control group. Both groups benefited substantially from naturally produced clear speech, and for both groups, the female talker evoked a larger clear speech benefit than the male talker. The clear speech benefit was consistent across groups; required no listener training; and, for a large proportion of the children with LDs, was sufficient to bring their performance within the range of the control group with conversational speech. Moreover, an acoustic comparison of conversational-to-clear speech modifications across the two talkers provided insight into the acoustic-phonetic features of naturally produced clear speech that are most important for promoting intelligibility for this population. PMID- 12647891 TI - Delayed predictive accuracy of narrative recall after traumatic brain injury: salience and explicitness. AB - Fifteen adults with traumatic brain injury (TBI) and 15 adults without brain injury listened to narratives, made delayed predictions of recall, and took a delayed recall test. Narrative questions differed by salience (main ideas, details) and explicitness (implied, stated) (R. H. Brookshire & L. E. Nicholas, 1993). TBI survivors recalled less than control participants regardless of question type. All participants recalled main ideas and implied information with greater accuracy than details and stated information. Predictive accuracy for recalling stated information was strong regardless of group. Participants were unable to predict recall for implied information. The materials-appropriate processing (MAP) hypothesis proposes that predictive accuracy is biased by text type (i.e., predictive accuracy for recalling main ideas should be higher than for details when learning narratives). However, there were no differences in predictive accuracy for recalling main ideas and details, with both groups predicting recall modestly well. Controlling for explicitness appears to be an important variable for future metamemory text studies. PMID- 12647892 TI - French-English bilingual children with SLI: how do they compare with their monolingual peers? AB - The goal of this study was to determine whether bilingual children with specific language impairment (SLI) are similar to monolingual age mates with SLI, in each language. Eight French-English bilingual children with SLI were compared to age matched monolingual children with SLI, both English and French speaking, with respect to their use of morphosyntax in language production. Specifically, using the extended optional infinitive (EOI) framework, the authors examined the children's use of tense-bearing and non-tense-bearing morphemes in obligatory context in spontaneous speech. Analyses revealed that the patterns predicted by the EOI framework were borne out for both the monolingual and bilingual children with SLI: The bilingual and monolingual children with SLI showed greater accuracy with non-tense than with tense morphemes. Furthermore, the bilingual and monolingual children with SLI had similar mean accuracy scores for tense morphemes, indicating that the bilingual children did not exhibit more profound deficits in the use of these grammatical morphemes than their monolingual peers. In sum, the bilingual children with SLI in this study appeared similar to their monolingual peers for the aspects of grammatical morphology examined in each language. These bilingual-monolingual similarities point to the possibility that SLI may not be an impediment to learning two languages, at least in the domain of grammatical morphology. PMID- 12647893 TI - Contralateral acoustic reflex thresholds for tonal activators using wideband energy reflectance and admittance. AB - The purpose of this study was to evaluate a new method for estimating the acoustic reflex threshold incorporating wideband (250-8000 Hz) measures of energy reflectance and admittance (M. P. Feeney & D. H. Keefe, 2001). The wideband technique incorporates both a correlation method to assess the pattern of the reflex-induced shifts in reflectance and admittance across frequency and a magnitude method to determine if the amplitude of the shifts exceeds baseline variability. Contralateral reflex thresholds for 1000- and 2000-Hz activators were obtained for 34 young adults with both the wideband method and a clinical method using a 226 Hz probe tone. Average reflex thresholds obtained with the new method were 12 to 13.7 dB lower than than obtained with the clinical method. When the bandwidth of analysis of admittance and reflectance responses was limited to 250 to 2000 Hz, the reduction in reflex thresholds was accompanied by the rejection of 96% of nonactivator-baseline responses as reflexes. The method holds promise for extending reflex threshold testing to patients with reflexes elevated beyond current equipment limits, for reducing the sound levels used in reflex testing, and for obtaining sensitive measures of reflex threshold in infants. PMID- 12647894 TI - An examination of changes in hearing-aid performance and benefit in the elderly over a 3-year period of hearing-aid use. AB - This brief report describes the changes in hearing-aid performance and benefit in 9 elderly hearing-aid wearers over a 3-year period following the hearing-aid fitting. Objective measures of hearing-aid performance included three measures of speech recognition: (a) the Nonsense Syllable Test (NST) presented at 65 dB SPL and a +8 dB signal-to-noise ratio (SNR), (b) the Connected Speech Test (CST) presented at 50 dB SPL in quiet, and (c) the CST presented at 65 dB SPL and a +8 dB SNR. Subjective, self-report measures of hearing-aid benefit included the Hearing Aid Performance Inventory (HAPI; B. E. Walden, M. E. Demorest, & E. L. Helper, 1984) and the Hearing Handicap Inventory for the Elderly (HHIE; I. Ventry & B. Weinstein, 1982). Performance and benefit measures were obtained at postfit intervals of 1 month, 6 months, 1 year, 2 years, and 3 years using a standardized measurement protocol. Individual data were evaluated using 95% critical differences established previously for each benefit measure and applied around the scores observed at the 1-month postfit interval. Little evidence was seen for systematic improvement in aided performance or benefit, consistent with that expected from acclimatization, in any participant or for any measure of benefit. PMID- 12647895 TI - Accuracy and variability of isochronous rhythmic timing across motor systems in stuttering versus nonstuttering individuals. AB - Ten adults who stutter and ten adults who do not stutter completed speech, orofacial nonspeech, and finger isochronous rhythmic timing tasks in a synchronization-continuation paradigm with auditory stimuli and with 450 ms, 650 ms, and 850 ms interstimulus onset intervals. Responses consisted of bilabial contact in the syllable /pa/ during a speech task, bilabial contact in an orofacial nonspeech task, and thumb-index finger contact in a finger movement task. Effector movements were transduced, and time points associated with minima in the derived lip or finger aperture signals were automatically extracted. Multiple analyses of timing accuracy and variability were completed for both the synchronization and continuation phases, including decomposition of total timing variance into central clock and motor implementation variance according to the Wing-Kristofferson model. The combined results from descriptive comparisons, statistical significance testing, and effect size computations suggest that the stuttering and nonstuttering participants showed highly similar levels of both timing accuracy and timing variability. This was true (a) for all three motor tasks, (b) at all movement rates, and (c) for synchronization as well as continuation movements. As one component of a systematic approach to investigating the role, if any, of timing difficulties in stuttering, these findings extend growing evidence that stuttering individuals do not differ from nonstuttering individuals in the ability to generate temporal movement patterns with a simple isochronous rhythm. We present some hypotheses about the implications that may follow from brain imaging and clinical neurological studies that have investigated the neural substrates recruited by this particular experimental task. PMID- 12647897 TI - Language and fluency characteristics of preschoolers' multiple-utterance conversational turns. AB - The present study examined language and fluency characteristics of single utterance (SU) and multiple-utterance (MU) conversational turns produced by 15 preschoolers who stutter and 15 age- and sex-matched preschoolers who do not stutter. Participants conversed with a parent in a play seating. Each interaction was videotaped and the participants' resultant utterances were transcribed and analyzed. Results indicated that the children's utterances from MU-turns typically served assertive functions and were significantly longer and more linguistically complex than their utterances from SU-turns. Neither group showed a significant difference in disfluency rate for length-matched utterances from MU and SU-turns. Similarly, there were no significant between-group differences in speaking-turn length or frequency of MU-turns. Although present findings do not support the hypothesis that MU-turns directly affect children's fluency, they do suggest that MU-turns are demanding for youngsters because they evoke relatively long and complex utterances. As such, conversational turn length seems to be an important variable for clinicians to consider when assessing and treating children who stutter. PMID- 12647898 TI - Acoustic and perceptual correlates of stress in nonwords produced by children with suspected developmental apraxia of speech and children with phonological disorder. AB - Previous research (L. Shriberg, D. Aram, & J. Kwiatkowski, 1997b, 1997c) has suggested that accuracy in producing linguistic stress reliably distinguishes between children with suspected developmental apraxia of speech (sDAS) and children with phonological disorder (PD). The current investigation tested this hypothesis by examining acoustic correlates of stress in trochaic (strong-weak) and iambic (weak-strong) nonwords produced by 5 children in each of these 2 groups. Four measures relating to stress production were examined: vowel duration, fundamental frequency (f0) at vowel midpoint, timing of the f0 peak relative to vowel onset, and intensity at vowel midpoint. In addition, perceptual judgments of accuracy of stress production were obtained. No group differences in the production of stress were found; however, listeners judged that the nonword repetitions of children with sDAS matched the target stress contour less often than did the repetitions of children with PD. Multiple regression analyses found that mean vowel duration, as well as the relative duration and relative f0 of stressed and stressless syllables, predicted listeners judgments of stress, although these variables only accounted for a small proportion of variance (21.8%). Thus, children with sDAS were able to produce acoustic differences between stressed tently perceptible to listeners. PMID- 12647896 TI - Respiratory sinus arrhythmia during speech production. AB - The amplitude of the respiratory sinus arrhythmia (RSA) was investigated during a reading aloud task to determine whether alterations in respiratory control during speech production affect the amplitude of RSA. Changes in RSA amplitude associated with speech were evaluated by comparing RSA amplitudes during reading aloud with those obtained during rest breathing. A third condition, silent reading, was included to control for potentially confounding effects of cardiovascular responses to cognitive processes involved in the process of reading. Calibrated respiratory kinematics, electrocardiograms (ECGs), and speech audio signals were recorded from 18 adults (9 men, 9 women) during 5-min trials of each condition. The results indicated that the increases in respiratory duration, lung volume, and inspiratory velocity associated with reading aloud were accompanied by similar increases in the amplitude of RSA. This finding provides support for the premise that sensorimotor pathways mediating metabolic respiration are actively modulated during speech production. PMID- 12647900 TI - Kinematic analyses of speech, orofacial nonspeech, and finger movements in stuttering and nonstuttering adults. AB - This work investigated the hypothesis that neuromotor differences between individuals who stutter and individuals who do not stutter are not limited to the movements involved in speech production. Kinematic data were obtained from gender and age-matched stuttering (n = 10) and nonstuttering (n = 10) adults during speech movements, orofacial nonspeech movements, and finger movements. All movements were performed in 4 conditions differing in sequence length and location of the target movement within the sequence. Results revealed statistically significant differences between the stuttering and nonstuttering individuals on several measures of lip and jaw closing (but not opening) movements during perceptually fluent speech. The magnitude of these differences varied across different levels of utterance length (larger differences during shorter utterances) and across different locations of the target movement within an utterance (larger differences close to the beginning). Results further revealed statistically significant differences between the stuttering and nonstuttering groups in finger flexion (but not extension) movement duration and peak velocity latency. Overall, findings suggest that differences between stuttering and nonstuttering individuals are not confined to the sensorimotor processes underlying speech production or even movements of the orofacial system in general. Rather, it appears that the groups show generalized differences in the duration of certain goal-directed movements across unrelated motor systems. PMID- 12647899 TI - Perception of vocal tremor. AB - Vocal tremors characterize many pathological voices, but acoustic-perceptual aspects of tremor are poorly understood. To investigate this relationship, 2 tremor models were implemented in a custom voice synthesizer. The first modulated fundamental frequency (F0) with a sine wave. The second provided irregular modulation. Control parameters in both models were the frequency and amplitude of the F0 modulating waveform. Thirty-two 1-s samples of /a/, produced by speakers with vocal pathology, were modeled in the synthesizer. Synthetic copies of each vowel were created by using tremor parameters derived from different features of F0 versus time plots of the natural stimuli or by using parameters chosen to match the original stimuli perceptually. Listeners compared synthetic and original stimuli in 3 experiments. Sine wave and irregular tremor models both provided excellent matches to subsets of the voices. The perceptual importance of the shape of the modulating waveform depended on the severity of the tremor, with the choice of tremor model increasing in importance as the tremor increased in severity. The average frequency deviation from the mean F0 proved a good predictor of the perceived amplitude of a tremor. Differences in tremor rats were easiest to hear when the tremor was sinusoidal and of small amplitude. Differences in tremor rate were difficult to judge for tremors of large amplitude or in the context of irregularities in the pattern of frequency modulation. These results suggest that difference limens are larger for modulation rates and amplitudes when the tremor pattern is complex. Further, tremor rate, regularity, and amplitude interact, so that the perceptual importance of any one dimension depends on values of the others. PMID- 12647901 TI - Acoustic startle responses and temperament in individuals who stutter. AB - Fourteen individuals who stutter and 14 individuals who do not stutter were presented with 10 bursts of white noise to assess the magnitude of their eyeblink responses as a measure of temperament. Both the magnitude of the eyeblink response to the initial noise burst and the mean of the 10 responses were significantly greater for the stuttering group. The Taylor-Johnson Temperament Analysis (R. M. Taylor & L P. Morrison, 1996) did not distinguish between the two groups, but informal follow-up statistics indicated that the Nervous subscale showed a significant group difference. Scores on this subscale were also significantly positively correlated with the magnitude of the startle response. A discriminant analysis demonstrated that although both the startle response and the nervous trait differentiated the two groups, the startle response measures were more powerful in making this differentiation. PMID- 12647902 TI - Posterior pharyngeal wall position in the production of speech. AB - The posterior pharyngeal wall has been assumed to be stationary during speech. The present study examines this assumption in order to assess whether midsagittal widths in the pharyngeal region can be inferred from measurements of the anterior pharyngeal wall. Midsagittal magnetic resonance images and X-ray images were examined to determine whether the posterior pharyngeal wall from the upper oropharynx to the upper laryngopharynx shows anterior movement that can be attributed to variables in speech: vowel quality in both English and Japanese; vowels versus consonants as classes of speech sounds; sustained versus dynamically produced speech; and isolated words versus sentences. Measurements were made of the distance between the anterior portion of the vertebral body and the pharyngeal wall. The first measurement was on a line traversing the junction between the dens and the body of the second cervical vertebra (C2). The next three measurements were on lines at the inferior borders of the bodies of C2, C3, and C4. The measurements showed very little movement of the posterior pharyngeal wall, none of it attributable to speech variables. Therefore, the position of the posterior pharyngeal wall in this region can be eliminated as a variable, and the anterior portion of the pharynx alone can be used to estimate vocal cavities. PMID- 12647904 TI - Avoiding painful blood sampling in neonates by transcutaneous bilirubinometry. AB - BACKGROUND: Transcutaneous bilirubinometry, Bilicheck (kindly provided by Oxygen Care Company, Dublin) is a non-invasive, screening method for determining bilirubin levels in neonates. It has not previously been validated in Irish babies. AIMS: To determine the clinical accuracy and precision of Bilicheck measurements in comparison to total serum bilirubin (TSB) measurements in Irish neonates, and to determine whether Bilicheck is a useful non-invasive screening method of avoiding blood tests in neonates. METHODS: Correlation data were obtained from 53 neonates between simultaneous TSB and Bilicheck readings. Bilicheck was used to screen 100 neonates who were assessed as requiring serum bilirubin measurement. TSB was only performed on neonates whose Bilicheck was above the phototherapy line of a recognised serum bilirubin chart. RESULTS: Bilicheck and TSB were significantly correlated (r=0.890). Bland and Altman analysis showed that on average, Bilicheck read 30 micromol/l lower than TSB. Of 100 jaundiced neonates screened by Bilicheck, blood sampling was avoided in 70 because Bilicheck did not meet phototherapy guidelines. CONCLUSIONS: There is a good correlation between TSB and Bilicheck, although the latter tends to under read by 30 micromol/l. Bilicheck may be a useful screening device to decrease the risks and discomfort associated with blood sampling in neonates. PMID- 12647905 TI - Critical appraisal of the application of carotid duplex scanning. AB - AIMS: To evaluate the indications for carotid arterial imaging in an open access vascular laboratory. To identify those symptoms predictive of > 50% stenosis of the carotid artery in order to reduce unnecessary imaging. To test the hypothesis that duplex scanning would not be of significant benefit in the management of those patients with ill defined symptoms. METHODS: We compared the outcome of carotid duplex scanning performed on 816 consecutive patients referred for a variety of clinical indications. The medical records of 816 patients were retrospectively analysed to identify the clinical indication for carotid duplex imaging over a three-year period (1997-9). RESULTS: The indications for duplex imaging were divided into two groups: definite carotid symptoms, n=350 (transient ischaemic attack n=205, cerebrovascular accident n=66, amaurosis fugax n=49, dysphasia n=30); and non-carotid symptoms, n=466 (dizziness n=63, syncope n=63, confusion n=20, vertigo n=10 and others n=310). Less than 5% of those with definite carotid symptoms and 2% of those with ill-defined symptoms had a stenosis > 80%. CONCLUSION: Regardless of symptoms, 14% and 2.9% of patients referred for carotid duplex imaging have a stenosis of > or = 50% and > or = 80%, respectively. Patients without definite carotid symptoms are of low priority for duplex imaging. PMID- 12647906 TI - Combination chemotherapy in the treatment of inoperable non-small cell lung cancer. AB - BACKGROUND: Chemotherapy is an established intervention in inoperable non-small cell lung cancer (NSCLC), yet few Irish patients receive this treatment. AIM: To determine survival, toxicity and radiological response following chemotherapy for NSCLC at our institution. METHODS: Retrospective audit of all patients receiving chemotherapy for histologically proven, inoperable NSCLC from January 1997 to December 2000. RESULTS: There were 80 treatment episodes in 77 patients, mean age 62 years. Forty-eight (60%) patients had locally advanced and 32 (40%) metastatic disease. Mitomycin, Ifosfamide, Carboplatin (MIC) and Paclitaxel/Carboplatin (PC) were the most commonly administered regimens. Median survival for locally advanced and metastatic disease was 13.9 months and 7.1 months respectively. Severe neutropenia and thrombocytopenia were each witnessed after less than 9% of cycles. Eleven (16.7%) patients had radiological response including 4 (6.1%) complete responses. CONCLUSION: Survival for inoperable NSCLC treated with chemotherapy was encouraging and achieved at low toxicity. PMID- 12647907 TI - Does prophylactic endoscopic sphincterotomy prevent recurrent biliary problems in patients with gallstones and a normal cholangiogram? AB - BACKGROUND: Endoscopic sphincterotomy (ES) is indicated in patients with confirmed bile duct stones at endoscopic retrograde cholangiopancreatography (ERCP). The role of ES in patients with suspected bile duct stones but a normal cholangiogram, in the prevention of recurrent biliary symptoms, when cholecystectomy is not planned, is unclear. AIM: To determine if prophylactic ES prevents further biliary problems in such patients. METHODS: Patients were identified with gallbladder stones presenting with jaundice, abnormal liver function tests (LFTs) or dilated bile ducts on ultrasound, in whom cholecystectomy was not planned and who had a normal cholangiogram at ERCP. Patients were followed-up to determine the frequency of recurrent biliary problems or repeat investigations. RESULTS: Forty-one patients were included, of whom 20 had an ES. The frequency of pre-ERCP features did not differ between the two groups. Median follow-up was 32 months (range 15-66). Post-ERCP recurrent abdominal pain (5 vs 3; p=0.39), jaundice (3 vs 1; p=0.28), pancreatitis (0 vs 1; p=0.32), and repeat ultrasound (2 vs 1; p=0.52), ERCP (1 vs 1; p=0.97) or cholecystectomy (2 vs 3, p=0.82) did not differ between the two groups. CONCLUSIONS: Patients with gallstones, suspected common bile duct (CBD) stones and a normal cholangiogram need not have a prophylactic sphincterotomy since there is no reduction in recurrent biliary problems and this potentially increases the morbidity. PMID- 12647909 TI - Prevalence of obstructive airflow limitation in Irish collegiate athletes. AB - BACKGROUND: Asthma prevalence in athletes is variable, depending on the sport and training conditions. AIMS: To investigate the prevalence of obstructive airflow limitation, asthma control and lung volume differences in a control group and five groups of collegiate athletes--outdoor field, indoor court, endurance, combat sports and swimming. METHODS: Three hundred and five athletes completed a respiratory questionnaire and performed spirometry during normal training conditions, to determine FEV1, FVC and PEFR pre- and at 5, 10 and 15 minutes post exercise. Between and within group analysis was carried out using single factor and repeated measures ANOVA. RESULTS: Previous diagnosis of asthma was reported by 24%, outdoor field sports 4.9%, indoor court sports 2.3%, endurance events 2.6%, combat sports 3%, swimming 6.9% and controls 4.3%. Mean FEV1 and FVC data, expressed as percentage of predicted, were greatest in swimming and lowest in controls. Swimming showed a significant rise in FEV1 post-exercise, indoor court and endurance sports demonstrated no change and outdoor field, combat and control groups showed a fall in FEV1 post-exercise. CONCLUSION: Warm humid training conditions appear protective against exercise-induced asthma. The greatest reductions in lung volumes occurred following high intensity exercise under cool conditions in the poorly aerobically conditioned. PMID- 12647908 TI - Mortality in epilepsy in the west of Ireland: a 10-year review. AB - BACKGROUND: Sudden and unexpected death is not an uncommon event in otherwise healthy epileptics. AIMS: To study the autopsied cases of sudden death in known epileptic patients in the West of Ireland. METHODS: A retrospective study was carried out which involved the review of 3,103 autopsy reports over a 10-year period in Galway University Hospital. RESULTS: Twenty-two cases were classified as sudden, unexpected death in epilepsy (SUDEP). Forty-five per cent had been found dead in bed. The mean age was 38 years and 68% were males. Of 16 cases with data about antiepileptic drug (AED) levels, 68% had absent or low levels at postmortem. Eighty-eight per cent of the cases had a past history of a generalised seizure. Pulmonary oedema was present in 86% of cases at postmortem. CONCLUSIONS: Compliance with treatment and frequent nightly observations to those at high risk of sudden death may help to minimise its incidence. The patients and their families should be aware of this potential outcome. PMID- 12647910 TI - An approach to the education about, and assessment of, attitudes in undergraduate medical education. AB - BACKGROUND: Attitudes have been shown to be important determinants of the quality and efficacy of medical care. There is little research on the education about, and assessment of, attitudes in undergraduate medical education. AIM: To describe the design and delivery of an attitude awareness workshop, and the associated objective structured clinical examination (OSCE) station used as the assessment tool. METHODS: Development, delivery, assessment and evaluation of an attitude awareness workshop were performed. RESULTS: Data are presented from 144 students. In 1999, the overall mean OSCE score was 62.44% (SD 7.6, n=73). The mean score for the attitude station was 57.97% (SD 12.9). In 2000, these figures were 67.11% (SD 8.3, n=71) and 73.75% (SD 10.8) respectively. In 1998/99, the average mark out of 10 for the educational quality of the attitude workshop was 6.6 and in 1999/00 this rose to 7.8. CONCLUSION: Development of both an educational and assessment programme concerning attitudes appears feasible. PMID- 12647911 TI - High dependency care: impact of lack of facilities for high-risk surgical patients. AB - BACKGROUND: The difference between the level of care available on general ward areas and intensive care units (ICUs) has resulted in the development of high dependency units (HDUs). AIMS: This study examined the current perioperative management techniques and clinical care settings of high-risk surgical patients in a hospital without a HDU. METHODS: A prospective audit of high-risk surgical patients was performed over an eight-week period. Using a pre-operative questionnaire, the anaesthetist categorised patient suitability for one of three post-operative care areas. In addition, desired and actual post-operative monitoring, pain management and organ support were indicated. RESULTS: Seven (25%) of 28 patients admitted to ICU were rated HDU suitable indicating inappropriate use of resources, while 27 (75%) of 36 patients admitted to the general ward were categorised as fulfilling HDU admission criteria. A total of 21/27 (78%) and 12/27 (44%) of this latter group had alteration of idealised post operative pain management and haemodynamic invasive monitoring plans respectively. CONCLUSIONS: The absence of an intermediate care area facility in a tertiary hospital without an acute pain team impacts on the type of perioperative care provided to patients and the optimal use of resources. PMID- 12647913 TI - Late presentation of small bowel obstruction following blunt abdominal trauma. AB - BACKGROUND: Motor vehicle accidents have increasingly become a major cause of serious blunt abdominal and chest injury, the pattern and mechanism of which has changed in recent years largely due to seatbelt legislation. AIM: A case of blunt abdominal and chest trauma is reported which resulted in a mesenteric tear--the small bowel subsequently herniated through and strangulated. CONCLUSION: This case highlights the need for clinical suspicion, serial physical examination and early surgery in the management of these injuries. PMID- 12647912 TI - Sigmoid volvulus: a 10-year-audit. AB - BACKGROUND: Chronic constipation in elderly, institutionalised patients is the leading cause of sigmoid volvulus in the developed world. Endoscopic deflation is associated with a 90% recurrence rate and a 35% mortality rate. AIMS: To review a 10-year experience of sigmoid volvulus and encourage more aggressive primary treatment. METHODS: A retrospective study was performed on 16 patients with sigmoid volvulus from 1992 to 1999. Patients were identified using the hospital inpatient enquiry (HIPE) data system. Demographics, clinical course, intervention, complications and outcome were recorded. RESULTS: The male:female ratio was 5:3 and mean age was 78 years (range 39-92). Fifty per cent had at least one risk factor: Parkinson's disease (n=3); multiple sclerosis (n=1); Alzheimer's disease (n=1); and hypokalaemia (n=3). Thirty-seven per cent were managed conservatively and 63% required surgical intervention. Mean time to surgery was 2.4 days. Operations performed were sigmoid colectomy (45%), Hartmann's procedure (33%) and total colectomy (22%). There was one post operative death from myocardial ischaemia. Mean duration of admission was 21 days. CONCLUSIONS: Endoscopic deflation of a sigmoid volvulus facilitates optimisation of cardiopulmonary co-morbidity in a high-risk group of patients. It converts an emergent to an elective procedure and minimises operative morbidity as a result. PMID- 12647915 TI - Medical statues--Irish and others. PMID- 12647914 TI - Conway Memorial Lecture 2002. The dyslipidaemia of diabetes: lessons in the pathogenesis of atherosclerosis. PMID- 12647916 TI - Gastric urease and peptic ulcer disease in Ireland in the 1940s--the Fitzgerald connection. PMID- 12647917 TI - Income inequality and health in contemporary Ireland. PMID- 12647918 TI - Mesoblastic nephroma in an adult: a case report. PMID- 12647919 TI - Pilomatrixoma of the spermatic cord. PMID- 12647920 TI - Pilonidal abscess of the sternum. PMID- 12647921 TI - How confident are you in your study? PMID- 12647922 TI - Adults with cerebral palsy: walking ability after progressive strength training. AB - The purpose of this study was to evaluate effects of a progressive strength training programme on walking ability in adults with cerebral palsy. Ten individuals with spastic diplegia (seven males, three females; mean age 31, range 23-44 years) participated twice a week over 10 weeks. Seven individuals with spastic diplegia (four males, three females; mean age 33, range 25-47 years) who did not receive strength training served as controls. All individuals were ambulatory but motor ability ranged from functional walkers to individuals who always required walking aids and used a wheelchair regularly. Significant improvements were seen in isometric strength (hip extensors p=0.006, hip abductors p=0.01), and in isokinetic concentric work at 30 degrees/s (knee extensors p=0.02) but not in eccentric work. Results also showed significant improvements in Gross Motor Function Measure (GMFM) dimensions D and E (p=0.005), walking velocity (p=0.005), and Timed Up and Go (p=0.01). There was no increase in spasticity for those who underwent strength training. Individuals in the control group did not show any significant improvement in any measured variable. The groups were small, however, and there was no significant difference between the groups in any measured variable. These findings suggest that a 10-week progressive strength training programme improves muscle strength and walking ability without increasing spasticity. PMID- 12647923 TI - Ischaemic stroke subtypes in children and adults. AB - Ischaemic stroke subtypes in children and adults were compared to determine the similarity in aetiologies. Thirty-six children (22 females, 14 males; median age 5 years 7 months, range 6 weeks to 15 years 10 months) and 50 adults (35 males, 15 females; median age 44 years, range 17 years 2 months to 49 years 11 months) who had presented with ischaemic stroke between 1995 and 2000, were categorized using a modified version of the Trial of Org 10172 in Acute Stroke Therapy (TOAST) classification. Proportions of patients in the subtypes of the TOAST classification system were significantly different in the two groups (chi2 test, p<0.01). The first three subtypes (large artery atherosclerosis, cardioembolic, and small vessel disease) accounted for the majority of adult strokes (27 of 50). In contrast, only three of 36 children were accounted for within these three subtypes. The majority of children (29 of 36) were classified within the 'other determined aetiology' subtype. Aetiology was undetermined in 12 of 50 adults compared with three of 36 children. Causes of ischaemic stroke in children and adults are distinct. A classification system for ischaemic stroke in children would be useful for collaborative studies. PMID- 12647924 TI - Prognosis of haemorrhagic stroke in childhood: a long-term follow-up study. AB - Little is known about long-term physical sequelae, cognitive functioning, and quality of life of children who have had a haemorrhagic stroke. Fifty-six patients (29 females, 27 males) under 16 years of age at time of the bleeding were studied. Mean age at time of bleeding was 7.7 years (range 1 month to 15.9 years). The primary site and cause of the bleeding at baseline were determined. Occurrences of death, re-bleedings, and seizures during follow-up were recorded. Patients who survived were invited for a follow-up examination including physical check-up, general screening of cognition, and an inventory of subjective health perception. Thirteen children died directly as a result of the haemorrhage; nine experienced a recurrent bleeding, which was fatal in three; six children developed epileptic seizures. At follow-up 36 of 56 patients were still alive. Mean follow-up time was 10.3 years (range 1.3 to 19.9 years) and mean age was 18.6 years (range 1.8 to 34.1 years). There was no patient lost to follow-up. Five patients declined to visit the hospital. In 15 out of 31 patients who could be examined, no physical impairment was observed, 11 had a hemiparesis of varying severity, and three had symptoms of cerebellar ataxia. One child had persisting tetraparesis and one persisting paraparesis. Signs of cognitive deficits were found in 15 patients. Of the children who survive haemorrhagic stroke, the physical and functional prognosis is relatively good, as almost all children were independent at follow-up. However, only a quarter of the surviving children had no physical or cognitive deficit after a mean follow-up period of 10 years. The majority had low self-esteem as well as emotional, behavioural, and health problems. PMID- 12647925 TI - Object release under varying task constraints in children with hemiplegic cerebral palsy. AB - Considerable attention has been given in recent years to fingertip force coordination during grasping and lifting small objects in children with cerebral palsy (CP). However, little is known about the children's ability to replace and release an object from grasp. The present study examined the coordination of fingertip forces during replacement and release of an object from grasp under varying task constraints in the involved hand of 15 children (10 males, five females, age range 7 to 14 years) with hemiplegic CP and in the non-dominant hand of 15 age-matched, typically-developing children (seven males, eight females). Participants released an object, instrumented with force transducers and held with a precision grip, onto a stable surface and onto an unstable surface (requiring higher accuracy) at self-paced and fast-as-possible speeds. Temporal and force measures were recorded and the dependent measures were tested using analyses of variance. Results showed that force coordination was impaired in children with hemiplegia, resulting in prolonged and uncoordinated replacement and release of the object (p<0.05). Differences between controls and children with hemiplegia were greater when speed and accuracy constraints were imposed (i.e. task performance was affected by these constraints to a greater extent in the children with CP, p<0.05). Impairments in temporal coordination of object release were also observed in the non-involved hand under all conditions (p<0.05). These results provide additional information about impaired hand function in children with hemiplegic CP. Clinical implications of these findings are discussed. PMID- 12647926 TI - Grip force parameters in precision grip of individuals with myelomeningocele. AB - The aim of this study was to characterize impairments of hand function in individuals with myelomeningocele (MMC) using a quantitative method. A grip-lift task was studied in 29 individuals with MMC (17 females, 12 males; mean age 12 years 4 months, SD 5 years 6 months; range 4 to 28 years) and 29 age- and sex matched control participants. A small object (weight 200 g) was grasped and lifted with a precision grip of the dominant hand and grip forces (GF) and time intervals were measured. Sensibility was examined with a two-point discrimination test. In those with MMC, the latency between GF onset of the thumb and index finger and ensuing preload duration were both significantly prolonged (p<0.01). Fingertip forces were excessively high and variable. Several parameters differed significantly between those with MMC and control individuals: GF peak, GF at start of loading, mean GF, and the SDs of the GF during static holding. Although the two-point discrimination task indicated deficits of sensibility, these did not correlate with grip force parameters. Results demonstrate that fine motor skills in patients with MMC are impeded by slowness and inadequate adjustments of manipulative forces. PMID- 12647927 TI - Plasma long-chain polyunsaturated fatty acids and neurodevelopment through the first 12 months of life in phenylketonuria. AB - The aims of the study were to examine the relationship between long-chain polyunsaturated fatty acid (LCPUFA) status at diagnosis of phenylketonuria (PKU) and neurodevelopment through the first 12 months of life, and to assess whether any difference exists between infants breastfed and bottlefed in the first days of life on the basis of LCPUFA status. Twenty infants with PKU were prospectively examined through the first year of life. Plasma fatty acids were measured in infants at diagnosis. Plasma phenylalanine levels were determined monthly. Main outcome measures were the Bayley Mental Developmental Index (MDI) and Psychomotor Developmental Index (PDI) at 5 and 12 months of age, and the visual function at 12 months, evaluated by P100 wave latencies with visual evoked potentials. A higher PDI score was associated with higher plasma arachidonic acid at diagnosis (adjusted correlation coefficient of PDI at 5 months, r=0.38, p=0.05). P100 wave latency at 15 minutes of arc (15') was associated with the early plasma arachidonic acid (adjusted r=-0.56, p=0.02) and phenylalanine levels at 12 months (adjusted r=0.22, p=0.05). No association was found between MDI score and any essential fatty acids. Breastfed infants exhibited higher plasma arachidonic acid (mean difference, delta, =3.4%; 95% confidence interval [CI]=1.2-5.6%) and shorter P100 wave latency at 15' (delta=-21 ms, 95%CI=-30 to -12) than bottlefed infants. Within the population of this study, a weak positive association has been found between plasma LCPUFAs at diagnosis (higher in breastfed infants) and neurodevelopmental indices through the first year of life. PMID- 12647929 TI - Visual and spatial working memory dissociation: evidence from Williams syndrome. AB - This study aimed at investigating the possible dissociation between visual and spatial working memory (WM) by means of two different experiments. In the first experiment, a WM test for visual material and for spatial information was given to a group of 202 normally developing children (chronological age 7 years 5 months, SD 1.6 years; 109 males, 93 females). Results document a different developmental trend in the two span tests. In the second experiment, we presented the same visual and spatial WM tests to a group of 13 individuals with Williams syndrome (WS), seven males and six females (chronological age mean 18 years 3 months, SD 5.1; mental age mean 7 years 2 months, SD 1.5 years), and to a control group of 26 typically developing children 14 males and 12 females matched for mental age (mean 7 years 2 months, SD 1.4). The results of this second experiment show that, on average, the spatial span obtained by individuals with WS was significantly lower than control participants, but visual span was comparable in the two groups. Our data support the hypothesis of a dissociation within the visuo-spatial sketch pad slave system in the WM model. PMID- 12647928 TI - Neurocognitive functioning in children with type-1 diabetes with and without episodes of severe hypoglycaemia. AB - Previous studies have shown that recurrent severe hypoglycaemia can cause long term cognitive impairment in children with type-1 diabetes, but the results are controversial, possibly due to the heterogeneity of samples and lack of comprehensive neuropsychological assessments of children. The aim of this study was to assess the effects of diabetes and severe hypoglycaemia on the neurocognitive functioning of children with a standardized, wide age-range neuropsychological test battery designed for the assessment of children. Eleven children with diabetes and a history of severe hypoglycaemia, 10 children with diabetes without a history of severe hypoglycaemia, and 10 healthy control children (a total of 31 children: 14 males and 17 females, age range 5 years 6 months to 11 years 11 months, mean 9 years 4 months, SD 1 year 11 months) were studied using the Wechsler Intelligence Scale for Children-Revised (WISC-R) and the NEPSY, a Developmental Neuropsychological Assessment. The NEPSY assessed development in attention and executive functions, language, sensorimotor functions, visuospatial processing, and learning and memory. Children with a history of severe hypoglycaemia had more neuropsychological impairments, more learning difficulties (as reported by parents), and needed more part-time special education than those in the other groups. Significant differences were found in verbal short-term memory and phonological processing. Results suggest that severe hypoglycaemia is a risk factor for learning due to deficits in auditory-verbal functioning. PMID- 12647930 TI - Skin-to-skin contact (Kangaroo Care) accelerates autonomic and neurobehavioural maturation in preterm infants. AB - The effects of mother-infant skin-to-skin contact (Kangaroo Care; KC) on autonomic functioning, state regulation, and neurobehavioural status was examined in 70 preterm infants, half of whom received KC over 24.31 days (SD 7.24) for a total of 29.76 hours (SD 12.86). Infants were matched for sex (19 males and 16 females in each group); birthweight (KC, 1229.95 g [SD 320.21]; controls, 1232.17 g [SD 322.15]); gestational age (GA) (KC, 30.28 weeks [SD 2.54]; controls, 30.19 weeks [SD 2.65]); medical risk; and family demographics. Vagal tone was calculated from 10 minutes of heart rate before KC and again at 37 weeks' GA. Infant state was observed in 10-second epochs during four consecutive hours before KC and again at 37 weeks' GA. Neurobehavioural status was assessed at 37 weeks' GA with the Neonatal Behavioral Assessment Scale (NBAS). Infants receiving KC showed a more rapid maturation of vagal tone between 32 and 37 weeks' GA (p=0.029). More rapid improvement in state organization was observed in KC infants, in terms of longer periods of quiet sleep (p=0.016) and alert wakefulness (p=0.013) and shorter periods of active sleep (p=0.023). Neurodevelopmental profile was more mature for KC infants, particularly habituation (p=0.032) and orientation (p=0.007). Results underscore the role of early skin-to-skin contact in the maturation of the autonomic and circadian systems in preterm infants. PMID- 12647931 TI - Modulation of spatial attention in a child with developmental unilateral neglect. AB - Attentional neglect of left space is one of the most striking acquired neurological disorders of adulthood. Recent evidence indicates a link between left spatial neglect and general right-hemisphere impairments in sustained attention and alertness. Poor sustained attention and alertness is also a central feature of other disorders, particularly childhood attention-deficit hyperactivity disorder (ADHD). Here we present the case of a 7-year-old male showing that frank neglect can be present in children with sustained attention problems without a clear aetiological event, or obvious structural brain abnormalities as indicated by a normal MRI. Experimental amelioration of the neglect through left-hand movement and externally alerting stimulation by uninformative sounds further suggest close similarities to the adult disorder. We suggest that such distortions of spatial attention may be more common in childhood than previously thought. PMID- 12647932 TI - The multifunctional or moonlighting protein CD26/DPPIV. AB - CD26/DPPIV can be considered a moonlighting protein because it is a multifunctional protein that exerts its different functions depending on cell type and intra- or extracellular conditions in which it is expressed. In the present review, we summarize all its known functions in relation to physiological and pathophysiological conditions. The protein is a proteolytic enzyme, receptor, costimulatory protein, and is involved in adhesion and apoptosis. The CD26/DPPIV protein plays a major role in immune response. Abnormal expression is found in the case of autoimmune diseases, HIV-related diseases and cancer. Natural substrates for CD26/DPPIV are involved in immunomodulation, psycho/neuronal modulation and physiological processes in general. Therefore, targeting of CD26/ DPPIV and especially its proteolytic activity has many therapeutic potentials. On the other hand, there are homologous proteins with overlapping proteolytic activity, which thus may prevent specific modulation of CD26/DPPIV. In conclusion, CD26/DPPIV is a protein present both in various cellular compartments and extracellularly where it exerts different functions and thus is a true moonlighting protein. PMID- 12647933 TI - Towards determining the differentiation program of antigen-presenting dendritic cells by transcriptional profiling. AB - Dendritic cells (DC) represent professional antigen-presenting cells that develop from hematopoietic progenitors through successive steps of differentiation. Employing DNA microarray technology, we analysed the specific changes in gene expression that occur when human progenitor cells differentiate into DC. CD34 progenitor cells were first amplified in vitro with stem cell factor (SCF), Flt3 ligand (FL), thrombopoietin and IL-6/soluble IL-6 receptor fusion protein, and cells were then induced to differentiate into DC with IL-4 and GM-CSF. DC maturation was induced by TNFalpha. Progenitor cells and DC were subjected to transcriptional profiling by DNA microarrays that represent 13000 human genes. Our analysis revealed specific changes in the expression of a large number of cell surface antigens including molecules involved in antigen uptake and processing, cell migration and antigen presentation. Genes encoding such molecules were upregulated during DC differentiation as were genes encoding cytokines, cytokine receptors, chemokines and chemokine receptors. Stem cell genes and genes related to the multilineage differentiation potential and proliferative state of progenitor cells were downregulated. Our analysis also provides information on the expression profiles of transcriptional regulators such as the NF-kappaB/rel and STAT transcription factors. Interestingly, NF kappaB/rel factors were found to be expressed in both progenitor cells and DC at similar levels and were induced by TNFalpha. In contrast, expression of STAT factors increased during DC differentiation and their expression was virtually unaffected by TNFalpha. PMID- 12647935 TI - Chemotactically directed redistribution of alpha-actinin precedes morphological polarization and reversal of polarity in human polymorphonuclear leucocytes (PMNs). AB - We have determined the temporal and spatial relationship between cell polarization and alpha-actinin localization by analysing the redistribution of alpha-actinin and F-actin in spherical PMNs developing polarity and in polarized cells reversing polarity following localized stimulation with chemotactic peptide using micropipettes. Initially spherical PMNs develop a one-sided accumulation of alpha-actinin before lamellipodia enriched in alpha-actinin are formed. In polarized cells, alpha-actinin is concentrated at the leading front. When polarity is reversed, alpha-actinin redistribution to the uropod precedes reversal of morphological polarity and formation of new lamellipodia at the uropod. Later, lamellipodia enriched in F-actin and alpha-actinin develop at the former uropod to form a new front. The data document that redistribution of alpha actinin is a very early event in the development of polarity, which precedes formation of lamellipodia. PMID- 12647934 TI - Kidney Na+,K(+)-ATPase is associated with moesin. AB - Na+,K(+)-ATPase is a ubiquitous plasmalemmal membrane protein essential for generation and maintenance of transmembrane Na+ and K+ gradients in virtually all animal cell types. Activity and polarized distribution of renal Na+,(+)-ATPase appears to depend on connection of ankyrin to the spectrin-based membrane cytoskeleton as well as on association with actin filaments. In a previous study we showed copurification and codistribution of renal Na+,K(+)-ATPase not only with ankyrin, spectrin and actin, but also with two further peripheral membrane proteins, pasin 1 and pasin 2. In this paper we show by sequence analysis through mass spectrometry as well as by immunoblotting that pasin 2 is identical to moesin, a member of the FERM (protein 4.1, ezrin, radixin, moesin) protein family, all members of which have been shown to serve as cytoskeletal adaptor molecules. Moreover, we show that recombinant full-length moesin as well as its FERM domain bind to Na+,K(+)-ATPase and that this binding can be inhibited by an antibody specific for the ATPase activity-containing cytoplasmic loop (domain 3) of the Na+,K(+)-ATPase alpha-subunit. This loop has been previously shown to be a site essential for ankyrin binding. These observations indicate that moesin might not only serve as direct linker molecule of Na+,K(+)-ATPase to actin filaments but also modify ankyrin binding at domain 3 of Na+,K(+)-ATPase in a way similar to protein 4.1 modifying the binding of ankyrin to the cytoplasmic domain of the erythrocyte anion exchanger (AE1). PMID- 12647936 TI - Characterization of heterokaryons between skeletal myoblasts and preadipocytes: myogenic potential of 3T3-L1 preadipocytes. AB - It has been shown previously that heterokaryons between myoblasts and non myogenic cells disturb myogenic differentiation (Hirayama et al. (2001); Cell Struct. Funct. 26, 37-47), suggesting that some myogenesis inhibitory factors exist in non-myogenic cells. Skeletal myoblasts and adipose cells are derived from a common mesodermal stem cell, indicating that both cells have a closer relationship in the developmental lineage than the other somatic cells. To investigate the functional relationship between myoblasts and adipose cells, heterokaryons between quail myoblasts and 3T3-L1 cells, a mouse preadipocyte cell line, were prepared and examined for characteristics of myogenic differentiation. Myogenic differentiation was inhibited in the heterokaryons between quail myoblasts and well-differentiated (adipocytes) 3T3-L1 cells. On the contrary, normal myogenic differentiation proceeded in the heterokaryons between quail myoblasts and undifferentiated (preadipocytes) 3T3-L1 cells. Further investigation showed that the mouse myogenin gene from 3T3-L1 cells was transactivated in the heterokaryons between quail myoblasts and undifferentiated 3T3-L1 cells. The results demonstrated that undifferentiated 3T3-L1 cells have no myogenesis inhibitory factors but acquire these during terminal differentiation into adipocytes. PMID- 12647937 TI - The economic impact of COPD in North America and Europe. Analysis of the Confronting COPD survey. Introduction. PMID- 12647938 TI - Economic analysis of the Confronting COPD survey: an overview of results. AB - An economic analysis of data from a large-scale international survey, Confronting COPD in North America and Europe, was conducted in seven countries (Canada, France, Italy, The Netherlands, Spain, the U.K and the U.SA.), to investigate the burden of chronic obstructive pulmonary disease (COPD). The results demonstrated the high economic impact of COPD on the healthcare system and society in each country. The mean annual direct costs of the disease were particularly high in the U.S.A. (US dollar 4119 per patient) and Spain (US dollar 3196 per patient) but relatively low in The Netherlands (US dollar 606) and France (US dollar 522). Lost productivity due to COPD had a particularly high impact on the economy in France, The Netherlands and the U.K, accounting for 67%, 50% and 41% of overall costs, respectively. The total societal cost of COPD per patient ranged from over US dollar 5646 in the U.S.A. to US dollar 1023 in The Netherlands. In five out of seven countries, the majority (52-84%) of direct costs associated with COPD were due to inpatient hospitalizations. As acute exacerbations of COPD are a key driver of secondary care costs, interventions aimed at preventing and treating exacerbations effectively could significantly reduce the economic impact of this disease. In all of the participating counties, COPD was underdiagnosed and undertreated. Between 9% and 30% of patients were undiagnosed despite having symptoms consistent with COPD, and up to 65% of patients did not receive regular prescribed medication. Patients reported poor symptom control and considerable use of healthcare resources. Therefore, reducing the burden of COPD will involve better evaluation and diagnosis of patients with COPD, as well as improved management of chronic COPD symptoms by healthcare professionals. The survey also demonstrated that the societal costs of COPD were 4-17 times higher in patients with severe COPD than in patients with mild COPD. Patients with comorbid conditions (accounting for 30-57% of patients in each country) were also particularly costly to society. These results suggest that a high priority should be given to interventions aimed at delaying the progression of disease, preventing exacerbations and reducing the risk of comorbidities, in order to alleviate the clinical and economic burden of COPD in North America and Europe. PMID- 12647939 TI - Economic analysis of the Confronting COPD survey: methodology. AB - Chronic obstructive pulmonary disease (COPD) is a condition characterized by progressive airflow limitation, which causes considerable morbidity and mortality worldwide. Yet the burden of COPD is poorly recognized, and the disease remains an inadequately managed health problem. Few studies have attempted to quantify the impact of the disease on patient health, the healthcare system and society as a whole. This provided the rationale for Confronting COPD in North America and Europe, the first large-scale international survey of the burden of COPD. This paper describes how quantitative measures of healthcare resource utilization and workplace productivity loss were derived from patient responses to the Confronting COPD survey, to investigate the country-specific impact of COPD on the healthcare system and society. The aim of this analysis is to inform countries of the economic impact of the condition, and demonstrate the need for better COPD treatment to improve health and reduce the sizeable burden of this disease. PMID- 12647940 TI - The burden of COPD in Canada: results from the Confronting COPD survey. AB - Chronic obstructive pulmonary disease (COPD) is a condition characterized by progressive airflow limitation, with symptoms of dyspnoea, cough, and sputum production. Aside from information on prevalence, mortality and hospital resource utilization arising from COPD in Canada, there is a lack of data on the impact of the disease on primary care healthcare resource utilization and the economic burden of the disease (i.e. direct and indirect costs). Canada is not unusual in this respect, as surprisingly few studies have attempted to quantify the impact of COPD on the healthcare system and society in other countries. In an attempt to address the need for information on the burden of COPD, an economic analysis of data from a large-scale international survey, Confronting COPD in North America and Europe, was conducted in Canada and six other countries. The results of the Canadian survey estimated the direct cost of the disease at CA dollar 1997.81 per patient, with over half of this due to inpatient hospitalizations. COPD also had an impact on the economy, with indirect costs amounting to CA dollar 1198.18, a third of the total per patient cost of COPD to society (CA dollar 3195.97). Reducing the impact of this disease will necessitate improvements to the way the disease is managed in primary care, as poor symptom control and frequent exacerbations are key drivers of hospital and other unscheduled care costs. Early diagnosis and the application of available but underused interventions (e.g. smoking cessation, inhaled drug therapies and pulmonary rehabilitation) could reduce the morbidity and costs of COPD in this country. PMID- 12647941 TI - The burden of COPD in France: results from the Confronting COPD survey. AB - Chronic obstructive pulmonary disease (COPD) is an obstructive lung disorder characterized by progressive airflow limitation that is not reversible or only partially reversible, including chronic bronchitis and emphysema. Confronting COPD in North America and Europe was the first large-scale international survey to attempt to quantify the country-specific burden of the disease, collecting data on clinical outcomes, healthcare resource utilization, and lost productivity, from patients and physicians in France and seven other countries. The economic analysis of the French survey results showed that patients with COPD required considerable utilization of healthcare resources, with annual direct costs estimated at Euro 530 per patient In addition, COPD-related illness or disability prevented many patients from working, with an estimated annual indirect cost of Euro 1078 per patient The survey suggested underdiagnosis and undertreatment of COPD by healthcare professionals, and patients reported poor symptom control. The cost of unscheduled care (Euro 151) was almost double the cost of scheduled visits to healthcare professionals (Euro 82). This suggests that improving the long-term management of chronic symptoms by healthcare professionals could reduce the burden of disease. As in other countries, the clinical management of COPD in France may be improved by following guideline recommendations for COPD treatment. These include smoking cessation at all stages of the disease, regular treatment of chronic symptoms with bronchodilators in dyspnoeic patients, and pulmonary rehabilitation. The results of the survey also showed that the societal cost of COPD was considerably greater in patients with severe disease (Euro 2882) compared with mild COPD (Euro 289). This suggests that interventions that could help delay the progression of COPD to the advanced stages of the disease (such as smoking cessation) could be of economic benefit. PMID- 12647942 TI - The burden of COPD in Italy: results from the Confronting COPD survey. AB - Chronic obstructive pulmonary disease (COPD) is a disease state characterized by airflow limitation that is not fully reversible, and progressive lung function decline. In Italy, an estimated 2.6 million men and women have COPD, and the disease causes around 18 000 deaths each year. In addition to mortality, morbidity from COPD results in substantial use of secondary healthcare resources. The burden of COPD in Italy may be due to the underdiagnosis of the disease by healthcare professionals, particularly in the early stages of the disease, and a lack of awareness among physicians of recommended treatment practices. In an attempt to address the need for information on the burden of COPD, a large-scale international survey, Confronting COPD in North America and Europe, assessed clinical outcomes, use of healthcare resources and loss of productivity in the workplace, and the economic cost of this disease in Italy and six other countries. In Italy, the economic analysis of the survey data showed that the mean annual cost of COPD to the healthcare system was Euro 1261.25 per patient Indirect costs were estimated at Euro 47.29 per patient, bringing the societal cost of the disease to Euro 1308.54 per patient. Three-quarters of the direct per patient cost of COPD in Italy were accounted for by inpatient hospitalizations (Euro 963.10), suggesting that interventions aimed at preventing exacerbations could alleviate the burden of COPD in Italy. The high impact of COPD on the Italian healthcare system is also likely to be a consequence of the underdiagnosis and undertreatment of the disease, suggesting that costs may be reduced by increasing the utilization of spirometry as a diagnostic tool, and improving physician adherence to treatment guidelines. Patients with severe COPD and other comorbidities showed higher costs (Euro 6366 and Euro 1861, respectively) than patients with mild disease (Euro 441) or no comorbidities (Euro 1021), underlying the importance of smoking cessation interventions to prevent disease progression. PMID- 12647943 TI - The burden of COPD in The Netherlands: results from the Confronting COPD survey. AB - Chronic obstructive pulmonary disease (COPD) is a disabling condition associated with progressive airflow limitation that is largely irreversible, with symptoms of dyspnoea, cough and sputum production. In The Netherlands, COPD tends to be underpresented to physicians, underdiagnosed and undertreated by healthcare professionals, and poorly recognized among the general population. Improving the diagnosis and management of COPD in this country may require raised awareness of the impact of the disease on society. This may be achieved by providing detailed information on the burden of COPD on the patient, healthcare system and the economy. This information has now become available from Confronting COPD in North America and Europe, the first international survey to quantify the country specific impact of the disease. An economic analysis of the results of the Dutch survey revealed the high cost of COPD to the healthcare system and society, with direct costs estimated at Euro 614 per patient Indirect costs amounted to Euro 410, bringing the annual per patient cost of COPD to Euro 1024. Around 50% of the cost of COPD to the healthcare system was for prescription medication, including symptomatic medication and treatment for underlying airway inflammation. This contributed to effective symptom control in many patients, as shown by the low utilization of unscheduled healthcare (inpatient hospitalizations, emergency room visits, or contacts with healthcare professionals). However, the survey suggested that there was scope for improvement in the understanding of the origin and consequences of this disease among the general public, and the way COPD is managed by healthcare professionals, in order to increase the proportion of patients who are diagnosed with COPD and treated in accordance with management guidelines. In addition, smoking cessation intervention at the early stages of the disease could help to reduce the high costs associated with severe COPD in this country. PMID- 12647944 TI - The burden of COPD in Spain: results from the Confronting COPD survey. AB - Chronic obstructive pulmonary disease is a smoking-related condition of progressive airflow obstruction, with disabling symptoms of chronic dyspnoea, cough and sputum production. In Spain, as in other countries worldwide, only a limited number of studies have attempted to quantify the impact of COPD on the patient, healthcare system and society. To obtain comprehensive information about the burden of this disease, an economic analysis of a large international survey, Confronting COPD in North America and Europe, was conducted. The results of the economic analysis of data from the Spanish survey sample estimated that the annual cost of COPD to the healthcare system was Euro 3238 per patient, with indirect costs amounting to Euro 300 per patient bringing the total societal cost of the disease to Euro 3538 per patient per year. A significant proportion of the economic burden of COPD on the Spanish healthcare system was associated with inpatient hospitalization (Euro 2708), which accounted for almost 84% of the total direct cost of the disease. As the major cause of inpatient hospitalization for COPD is acute exacerbations, these results highlight the need for interventions in the outpatient setting to prevent exacerbations in Spain. The impact of COPD on the healthcare system may also be due to the underdiagnosis and treatment of COPD, suggesting that costs may be reduced by improving the diagnosis and treatment of the disease in primary care. The sub-analysis of costs from the survey showed that patients with severe COPD were associated with considerably higher total societal costs than patients with mild disease (Euro 9850 versus Euro 1316 per patient). Therefore, introducing interventions to reduce the progression to severe COPD could also reduce the impact of the disease. PMID- 12647945 TI - The burden of COPD in the U.K.: results from the Confronting COPD survey. AB - Chronic obstructive pulmonary disease (COPD) is a condition characterized by progressive airflow limitation and decline in lung function. As seen in other developed countries throughout the world, COPD in the U.K is associated with considerable mortality, and morbidity from the disease places a significant burden on the healthcare system and society. Despite the obvious burden of COPD in this country, there is a lack of recognition of COPD among the general public. Healthcare professionals may also fail to recognize the burden of disease, as shown by underdiagnosis and inadequate managemen. A key step in increasing awareness of the burden of COPD is obtaining comprehensive information about the impact of the disease on patients, the health service and society. The large scale international survey, Confronting COPD in North America and Europe, aimed to address this need for information, by interviewing patients and physicians in eight countries. An economic analysis of patient responses to the survey in the U.K showed that COPD places a high burden on the healthcare system and society with annual direct costs estimated at pounds 819.42 per patient, and indirect cost at pound 819.66 per patient resulting in total per patient costs of pound 1639.08. The cost impact of the disease was particularly marked in secondary care, as a result of inpatient hospitalizations, amounting to 54% of direct costs. These results suggest that reducing patient requirement for hospital care could alleviate the burden of COPD on the U.K. healthcare system. This will require considerable improvements to the way the disease is managed by healthcare professionals in primary care, with earlier diagnosis and the use of interventions aimed at preventing exacerbations and delaying the progression of disease. PMID- 12647946 TI - The burden of COPD in the U.S.A.: results from the Confronting COPD survey. AB - Chronic obstructive pulmonary disease (COPD) is a progressive disorder of airflow limitation that is not fully reversible, with disabling symptoms including chronic cough and dyspnoea. Although a number of studies in the U.S.A. have assessed the impact of COPD on the healthcare system and society, data on healthcare resource utilization (particularly outpatient services and medication use) in patients with mild to moderate COPD, or patients who meet symptom criteria for COPD but have not received this diagnosis, are limited or unavailable. To fill gaps in current knowledge about the impact of this disease, an economic analysis was conducted on the data collected from patients enrolled in the U.S.A. sample of Confronting COPD in North America and Europe, the first large-scale international survey of the burden of the disease. The annual cost of healthcare resource utilization was estimated at US dollar 4119 per patient with COPD, with indirect (non-medical care) costs amounting to US dollar 1527 per patient. The annual estimated societal cost was therefore US dollar 5646 per patient. The majority of disease costs in the survey were associated with inpatient hospitalizations (US dollar 2891). The results of the survey suggest that interventions that improve COPD outcomes by decreasing symptoms and preventing acute exacerbations could substantially decrease the costs associated with this disease. PMID- 12647947 TI - Biochemistry, genetics, and applications of exopolysaccharide production in Streptococcus thermophilus: a review. AB - Many strains of Streptococcus thermophilus synthesize extracellular polysaccharides. These molecules may be produced as capsules that are tightly associated with the cell, or they may be liberated into the medium as a loose slime (i.e., "ropy" polysaccharide). Although the presence of exopolysaccharide does not confer any obvious advantage to growth or survival of S. thermophilus in milk, in situ production by this species or other dairy lactic acid bacteria typically imparts a desirable "ropy" or viscous texture to fermented milk products. Recent work has also shown that exopolysaccharide-producing S. thermophilus can enhance the functional properties of Mozzarella cheese, but they are not phage-proof. As our understanding of the genetics, physiology, and functionality of bacterial exopolysaccharides continues to improve, novel applications for polysaccharides and polysaccharide-producing cultures are likely to emerge inside and outside the dairy industry. This article provides an overview of biochemistry, genetics, and applications of exopolysaccharide production in S. thermophilus. PMID- 12647948 TI - Increase of viability of entrapped cells of Lactobacillus delbrueckii ssp. bulgaricus in artificial sesame oil emulsions. AB - A technique was developed to protect lactic acid bacteria (Lactobacillus delbrueckii ssp. bulgaricus) against simulated gastrointestinal conditions by encapsulation of bacterial cells within artificial sesame oil emulsions. Purified sesame oil bodies consisting of approximately 99% oil, 0.5% phospholipid, and 0.5% protein were decomposed by heating at 70 degrees C for 1 h. The bacteria cultured in nonfat milk were encapsulated in artificial oil emulsions constituted with decomposed sesame oil bodies and excess sesame or vegetable cooking oil. Viability of bacteria in storage at 4 degrees C for 16 d was substantially elevated from 0.023 to 5.45% after encapsulation. Compared with free cells, the entrapped bacteria demonstrated a significant increase (approximately 10(4) times) in survival rate when subjected to simulated high acid gastric or bile salt conditions. The results indicate that artificial sesame oil emulsion may serve as an effective biocapsule for encapsulation of bacteria in dairy products. PMID- 12647949 TI - The impact of fermentation and in vitro digestion on the formation of angiotensin I-converting enzyme inhibitory activity from pea and whey protein. AB - Pea and whey protein were fermented by Lactobacillus helveticus and Saccharomyces cerevisiae in monoculture and in combination at 28 and 37 degrees C in order to release angiotensin-I-converting enzyme (ACE) inhibitory peptides. The fermentation products were subjected to in vitro gastrointestinal digestion, and the digests of nonfermented samples served as controls. After fermentation, the ACE inhibitory activity (%) increased by 18 to 30% for all treatments, except for the fermentations of whey protein with Saccharomyces cerevisiae at 28 degrees C, where no significant change was observed. After digestion, however, both fermented and nonfermented samples reached maximum ACE inhibitory activity. The whey digests tended to have lower (50%) inhibitory concentrations (IC50; 0.14 to 0.07 mg/ml), hence, higher ACE inhibitory activity, than the pea digests (0.23 to 0.11 mg/ml). The nonfermented whey protein digest showed the highest ACE inhibitory activity of all. For pea protein, the nonfermented sample had the lowest IC50 value. These results suggest that in vitro gastrointestinal digestion was the predominant factor controlling the formation of ACE inhibitory activity, hence, indicating its importance in the bioavailability of ACE inhibitory peptides. PMID- 12647951 TI - The effect of compression, stretching, and cooking temperature on free oil formation in mozzarella curd. AB - The effect of the extent and rate of compression and stretching on free oil formation in Mozzarella cheese curd was investigated at 55, 65, and 75 degrees C. Confocal laser scanning microscopy was used to determine the maximum cross sectional diameter, cross-sectional area, elongation factor (maximum divided by minimum cross-sectional diameter), and circularity of fat globules in the cheese curd at the different temperatures, and after stretching or compression. Free oil was not significantly affected by the rate of biaxial compression from 50 to 2000 mm/min at 65 degrees C, the rate of tensile stretching from 1000 to 2500 mm/min at 60 degrees C, or the extent of biaxial compression from 40 to 80% of the original height at 1000 mm/min and 65 degrees C. Increasing the rate of stretching from 1000 to 2500 mm/min increased the elongation factor from 1.91 to 2.61. Cross-sectional area, maximum diameter, and circularity were not affected by the rate of biaxial compression. The extent of curd compression had no effect on the milk fat globule size and shape. Increasing the extent of stretching at 60 degrees C and at 1000 mm/min increased the free oil content (on a fat basis) from 23.8% (curd stretched by 1.4x) to 32.3% (stretched by 4.6x) and the elongation factor of the globules, but did not affect any of the other globule parameters. Increasing the temperature of the cooking-stretching water increased the cross sectional area, diameter of the globules, and free oil content from 24.1% at 55 degrees C to 34.5% at 75 degrees C for curd compressed to 50% height at 1000 mm/min. PMID- 12647950 TI - Determining flavor and flavor variability in commercially produced liquid cheddar whey. AB - Dried whey and whey protein are important food ingredients. Functionality of whey products has been studied extensively. Flavor inconsistency and flavors which may carry through to the finished product can limit whey ingredient applications in dairy and nondairy foods. The goal of this research was to determine the flavor and flavor variability of commercially produced liquid Cheddar cheese whey. Liquid Cheddar cheese whey from five culture blends from two different stirred curd Cheddar cheese manufacturing facilities was collected. Whey flavor was characterized using instrumental and sensory methods. Wide variation in whey headspace volatiles was observed between different manufacturing facilities (P < 0.05). Hexanal and diacetyl were two key volatiles that varied widely (P < 0.05). FFA profiles determined by solid-phase microextraction and degree of proteolysis of the whey samples were also different (P < 0.05). Differences in whey flavor profiles were also confirmed by descriptive sensory analysis (P < 0.05). Differences in liquid whey flavor were attributed to differences in milk source, processing and handling and starter culture blend. The flavor of liquid Cheddar cheese whey is variable and impacted by milk source and starter culture rotation. Results from this study will aid future studies that address the impact of liquid whey flavor variability on flavor of dried whey ingredients. PMID- 12647952 TI - Detection limits of antimicrobials in ewe milk by delvotest photometric measurements. AB - The Delvotest method detection limits per manufacturer's instructions at a fixed reading time of 3 h for 24 antimicrobial agents were determined in ewe milk by photometric measurement. For each drug, eight concentrations were tested on 20 ewe milk samples from individual ewes. Detection limits, determined by means of logistic regression models, were (microg/kg): 3, amoxycillin; 2, ampicillin; 18, cloxacillin; 1, penicillin "G"; 34, cefadroxil; 430, cephalosporin "C"; 40, cephalexin; 20, cefoperazone; 33, Ceftiofur; 18, cefuroxime; 6100, streptomycin; 1200, gentamycin; 2600, neomycin; 830, erythromycin; 100, tylosin; 180, doxycycline; 320, oxytetracycline; 590, tetracycline; 88, sulfadiazine; 44, sulfamethoxazole; 140, sulfametoxypyridazine; 48, sulfaquinoxaline; 12,000, chloramphenicol; and 290, trimethoprim. Whereas the beta-lactam antibiotics, sulphonamides, and tylosin were detected by Delvotest method at levels equal to those of maximum residue limits, its sensitivity needs to be enhanced to detect aminoglycosides, tetracyclines, streptomycin, chloramphenicol, and trimethoprim residues in ewe milk or to develop an integrated residue detection system for ewe milk with different sensitive microorganisms for each group of antiinfectious agents. PMID- 12647953 TI - Isolation and structural elucidation of a peptide derived from Edam cheese that inhibits beta-lactoglobulin transport. AB - A peptide that inhibits beta-lactoglobulin absorption in an in vitro Caco-2 cell model was isolated from Edam cheese. By 1H-NMR and N-terminal amino acid analyses, the active compound was identified as Asp-Lys-Ile-His-Pro-Phe. The sequence of the hexapeptide is the same as the amino acid sequence of residues 47 52 of beta-casein. The hexapeptide shows remarkable inhibition of beta lactoglobulin transport at a concentration of 10(-7)M. The possibility exists that this peptide can be applied practically to the prevention of milk-sensitive allergy. PMID- 12647954 TI - Transfer of protein from milk to cheese. AB - This report concerns measurement of paracasein in milk and transfer of protein from milk to cheese. In the main experiment, two vats of Cheddar cheese were made from each of 11 lots of milk from one large herd over a period of 7 mo. Exclusion of solutes from moisture in paracasein micelles in milk and cheese was central to estimation of paracasein and to the transfer of protein from milk to cheese and whey. Solute-exclusion by paracasein and its changes during cheesemaking could be visualized by considering paracasein micelles to be a very fine sponge. The sponge excludes solutes, especially the large solutes like whey proteins. The sponge shrinks during cheesemaking and expels solute-free liquid, thereby slightly diluting the whey surrounding the micelles inside the curd. Paracasein N in milk was calculated as the difference between total milk N and rennet whey N, the latter adjusted to its level in milk. Adjustment used appropriate solute exclusion factors (h) of the protein fractions of whey and 1.08 for paracasein and associated salts. They were combined to give a factor Fpc, which adjusted the level of rennet whey N to its level in milk: 1.001 x (1 - 1.01 x FM/100 - Fpc x pc/100), where FM = fat in milk, pc = estimated paracasein, and 1.001 = dilution of milk by chymosin and CaCl2. The mean Fpc was 3.03. Differences in values were small among different procedures for calculating paracasein, but they are considered to be important because they represent biases, which, in turn, are important in analyses commercially. We conclude that solute exclusion by moisture in paracasein must have decreased during cheesemaking because the ratio of moisture to paracasein in the final cheese was 1.5, much less than the h of 2.6 for serum proteins by paracasein. Release of solute-excluding moisture from paracasein during cooking was likely the reason for lower total N in cheese whey than in the rennet whey in the paracasein analysis. Paracasein, estimated to be in cheese, curd fines, salted whey, and whey during cheddaring, was 98.21, 0.20, 0.25 and 0.19%, respectively, of the paracasein in milk for a total of 98.85% (SD of 22 vats = 0.46); the location of the missing paracasein is not known. On the other hand, recovery of milk N in cheese and wheys was 99.92% (SD = 0.37%). Nitrogen in paracasein and its hydrolysis products in cheese was estimated to be 98.51% of total cheese N. Proteose-peptone from milk appeared not to be included with the paracasein in appreciable amounts. Some was apparently included with denatured serum proteins during Rowland fractionation of whey, perhaps as a coprecipitate. Measured paracasein would include fat globule membrane proteins in milk containing fat, and denatured whey proteins in heated milks. It was concluded that the method of measurement and the associated calculations are integral parts of the definition and quantification of paracasein in milk. PMID- 12647955 TI - Relationships between flavoring capabilities, bacterial composition, and geographical origin of natural whey cultures used for traditional water-buffalo mozzarella cheese manufacture. AB - Natural whey cultures (NWC) (n = 29) used for traditional water-buffalo Mozzarella cheese manufacture and arising from different geographical areas of production were characterized and grouped on the basis of their capability to develop neutral volatile compounds and according to their microbial diversity as revealed by molecular analysis. The flavoring properties of NWC were studied in dairy microcosms resembling the specific technological procedure used in the traditional water-buffalo Mozzarella cheese-making. Neutral volatile compounds were identified by high-resolution gas chromatography (HRGC)-mass spectrometry analysis while information on the microbial diversity occurring in the NWC was retrieved by PCR-denaturing gradient gel electrophoresis (DGGE) analysis of 16S rDNA after direct DNA extraction. Neoformation volatile substances (n = 27) were found; 23 were identified and some of them recognized as odor-conferring molecules. Eight different bands, referable to eight microbial species, were obtained by PCR-DGGE analysis of the NWC. Statistical analyses were applied to PCR-DGGE and HRGC data. Interestingly, the flavoring capabilities and the microbial diversity of the NWC proved to be closely linked and both related to the geographical origin of the NWC. These results suggested a possible use of the molecular characterization of the dairy products to support the traceability criteria of typical dairy products like water-buffalo Mozzarella cheese. PMID- 12647956 TI - 5'-flanking regions of camel milk genes are highly similar to homologue regions of other species and can be divided into two distinct groups. AB - The concentrations of individual casein and whey proteins in camel milk differ markedly to respective protein concentrations in bovine milk. The ratio of beta casein to kappa-casein is considerably higher in camel milk. beta-Lactoglobulin is absent, but whey acidic protein and peptidoglycan recognition protein have been detected. Genomic sequences upstream to milk-protein genes, which are known to regulate the expression of milk proteins to a great extent, were determined for 10 camel milk-protein genes and compared to respective sequences in other mammals. Multiple sequence alignment showed closest relationships to homologous sequences from other mammals. Comparison of milk protein regulative regions revealed two distantly related groups with pronouncedly different transcription factor site probabilities. The GC-content in sequences of the first group was considerably higher than in sequences of the second group and combined occurrence of CAAT and TATAA boxes was rare, suggesting that the first group represented mostly the housekeeping gene type, probably regulated by cellular signal transduction pathways, whereas the second group helped to regulate genes specifically expressed in terminally differentiated cells of the lactating alveolar epithelium. A core region of the composite response element, which primarily controls milk protein gene activity, was found by a search for elements conserved within all 5'-flanking sequences analyzed, and it is assumed, that the presence of this element determines gene expression in the lactating mammary gland, and binding sites for general activator and repressor factors, surrounding the milk protein gene specific element, are important for regulation of gene activity. PMID- 12647957 TI - Growth hormone and milking frequency act differently on goat mammary gland in late lactation. AB - In ruminants, milk yield can be affected by treatment with growth hormone (rbGH) and/or changes in frequency of milking. Frequent milkings encourage the maintenance of lactation, whereas infrequent milkings result in mammary involution. Our objective was to evaluate the influence of rbGH treatment and milking frequency on mammary gland morphology and milk composition. After adaptation to twice-daily milkings, six Saanen goats in late lactation were milked once daily from one udder-half and thrice-daily from the other udder-half. Concurrently, three of the six goats received daily injections of rbGH. After 23 d of treatment, milking frequency significantly affected milk yield (+8% vs. -26% for thrice- vs. once-daily milking). Additionally, treatments of rbGH increased milk yield from thrice-daily milked udder-halves (+19%), but failed to abate the reduction in milk yield from once-daily milked udder-halves (-31%). Mammary glands were heavier in the frequently milked udder-halves and in GH-treated goats. Based on histological and DNA analysis of mammary tissues, it was determined that milking frequency clearly affected epithelial cell numbers and alveolar diameter, whereas rbGH induced a potential cell hypertrophy and only a tendency to increase and/or maintain the mammary cell number. RNA concentration and kappa casein gene expression were not affected by treatments. In udder-halves milked once-daily, low casein:whey protein ratios, high Na+:K+ ratios, and high somatic cell counts (SCC) were indicative of changes in epithelial permeability, which rbGH treatment facilitated. The present data suggest that milking frequency and exogenous treatments of rbGH use different cellular mechanisms to influence mammary gland morphology and milk production. PMID- 12647958 TI - Effects of three types of free-stall surfaces on preferences and stall usage by dairy cows. AB - One important criterion in choosing appropriate housing systems for dairy cattle is that the freestall provides a comfortable surface for the cow. This paper describes two experiments testing the effects of commonly used lying surfaces on stall preference and stall usage by Holstein cows. In both experiments, 12 cows were housed individually in separate pens. Each pen contained three free stalls with a different surface: deep-bedded sawdust, deep-bedded sand, and a geotextile mattress covered with 2 to 3 cm of sawdust. The animals were restricted to each surface in turn, in a random order for either 2 (Experiment 1) or 3 d (Experiment 2). Both before and after this restriction phase, the animals were allowed access to all three surfaces, and preference was determined, based on lying times. Of the 12 cows used in Experiment 1, 10 preferred sawdust before and nine after the restriction phase. During the restriction phase, average lying times and number of lying events during the restriction phase were significantly lower for the sand-bedded stalls (P < or = 0.05), and standing times were higher on mattresses (P < or = 0.05), compared with sawdust. Although these cows had some experience with all three surfaces during the experiment, they had been housed in sawdust bedded stalls during their previous lactation. Cows used in Experiment 2 had spent their previous lactation in sand bedded stalls. In this experiment, about half the cows preferred sand and half sawdust, after the restriction phase. During the restriction phase of experiment, lying times and number of lying events were lower, and standing times were higher when the animals were restricted to the mattresses compared to either sand or sawdust (P < or = 0.05). These results indicate that (1) free stall surface can affect both stall preferences and stall usage, and (2) mattresses are less preferred. PMID- 12647959 TI - Influence of pulsation rate on udder health and teat thickness changes in dairy ewes. AB - In this work on machine milking of ewes, pulsation rates of 120 and 180 cycles per min were compared, both with a pulsation ratio of 50:50 and a vacuum level of 36 kPa, comparing intramammary infection (IMI), somatic cell count (SCC) and teat end thickness changes. To this end, two groups of 20 Manchega ewes were used in a crossover experimental design with two experimental periods of 24 d for each. Bacterial exposure of all teats was increased by dipping them in a suspension of Staphylococcus simulans at four consecutive milkings of each period. Pulsation rate of 180 cycles per min, compared with 120 cycles per min, had no negative effect upon new IMI (11 and 16% of ewes infected, respectively) and SCC. No teat end lesions were observed in those animals milked with the two pulsations assayed. Also, teat thickness changes (-0.38 and -0.36 mm at 120 and 180 cycles per min, respectively) were not affected significantly. Finally, in absence of IMI, the two pulsation rates assayed did not affect the SCC. PMID- 12647960 TI - Gene expression of immunologically important factors in blood cells, milk cells, and mammary tissue of cows. AB - Cytokines, eicosanoids, and lactoferrin are involved in the mammary gland's immune response to invading microorganisms. The goal of this work was to investigate the synthesis of these immunologically important factors in somatic milk cells, blood cells, and mammary tissue of cows with different somatic cell count levels, i.e., different immunological activity. On the level of mRNA expression, the cytokine tumor necrosis factor alpha (TNFalpha), lactoferrin (Lf), and specific key enzymes of leukotriene and prostaglandin biosynthesis, 5 lipoxygenase (5-LO), and cyclooxygenase-1 (COX-1) and -2 (COX-2), respectively, were determined. All 15 experimental cows were clinically healthy with no visible mammary disease. Eight cows were defined as control group with all quarters <150,000 cells/ml (C), whereas seven cows had partially elevated quarter somatic cell counts, with at least one quarter >150,000 cells/ml (H) and one quarter <150,000 cells/ml (L). Total quarter milk from one quarter of control group and from two quarters of cows with partially elevated cell counts (one of H and one of L) was collected at one milking and a blood sample was taken simultaneously. In addition, mammary tissue samples were taken from the respective quarters on the following day during slaughter. Total RNA from milk, blood, and tissue cells was isolated and reverse transcription and quantitative polymerase chain reaction was carried out. All factors investigated were not significantly different between groups in blood cells and between C and L quarters in milk cells and mammary tissue. TNFalpha and COX-2 mRNA expression was higher in milk cells and mammary tissue of H than in L quarters, except for COX-2 in mammary tissue. Generally, TNFalpha and COX-2 showed their highest expression in milk cells, 5-LO in blood cells, whereas lactoferrin was mainly expressed by the mammary tissue. COX-1 was similarly expressed in all tested samples. PMID- 12647961 TI - Effect of method of application of a fibrolytic enzyme product on digestive processes and milk production in Holstein-Friesian cows. AB - Four multiparous cows with cannulas in the rumen and proximal duodenum were used in early lactation in a 4 x 4 Latin square experiment to investigate the effect of method of application of a fibrolytic enzyme product on digestive processes and milk production. The cows were given ad libitum a total mixed ration (TMR) composed of 57% (dry matter basis) forage (3:1 corn silage:grass silage) and 43% concentrates. The TMR contained (g/kg dry matter): 274 neutral detergent fiber, 295 starch, 180 crude protein. Treatments were TMR alone or TMR with the enzyme product added (2 kg/1000 kg TMR dry matter) either sprayed on the TMR 1 h before the morning feed (TMR-E), sprayed only on the concentrate the day before feeding (Concs-E), or infused into the rumen for 14 h/d (Rumen-E). There was no significant effect on either feed intake or milk yield but both were highest on TMR-E. Rumen digestibility of dry matter, organic matter, and starch was unaffected by the enzyme. Digestibility of NDF was lowest on TMR-E in the rumen but highest postruminally. Total tract digestibility was highest on TMR-E for dry matter, organic matter, and starch but treatment differences were nonsignificant for neutral detergent fiber. Corn silage stover retention time in the rumen was reduced by all enzyme treatments but postruminal transit time was increased so the decline in total tract retention time with enzymes was not significant. It is suggested that the tendency for enzymes to reduce particle retention time in the rumen may, by reducing the time available for fibrolysis to occur, at least partly explain the variability in the reported responses to enzyme treatment. PMID- 12647962 TI - Effect of quantity, quality, and length of alfalfa hay on selective consumption by dairy cows. AB - Twenty-four lactating Holstein cows were used in a replicated 6 x 6 Latin square design. Experimental periods were 6 or 7 d. Cows were housed in tie-stalls, and diets were fed ad libitum twice daily at 1100 and 1600 h. Diets contained 60% concentrate and either 40% alfalfa hay or 20% alfalfa hay and 20% alfalfa silage (dry matter basis). The effect of quantity, quality, and length of hay on sorting behavior was determined. Treatments consisted of 20% lower or higher quality long alfalfa hay, 20% lower or higher quality chopped alfalfa hay, and 40% lower or higher quality chopped alfalfa hay. Variation of sorting among cows was also determined. Particle size distribution of samples of as-fed total mixed rations and orts were determined using the Wisconsin particle size separator. Screens have square holes with diagonals of 26.9, 18, 8.98, 5.61, and 1.65 mm (screens Y1 to Y5, respectively). Sorting was calculated as the actual intake of each fraction expressed as a percentage ofthe predicted intake. Increasing the proportion of dry hay increased sorting. Quality of alfalfa hays that were offered did not affect sorting activity. Feeding long alfalfa hay increased selective consumption of fine particles. However, feeding long alfalfa hay also increased intake of longer particles because a higher percentage of long particles was offered. Across treatments, animals consistently sorted against longer particles in favor of finer particles. In particular, intake of Y1 as a percentage of the predicted intake was the most variable. Average Y1 intake, across the six treatments for each cow, was between 60 and 70% of predicted intake for four cows, 71 to 80% for 11 cows, 81 to 90% for five cows, 91 to 100% for two cows, and 101 to 110% for two cows. On one diet a cow failed to consume any of the Y1 portion of the total mixed ration. This variation among animals in sorting of very long feed particles may have practical significance. PMID- 12647963 TI - Fibrolytic enzymes and parity effects on feeding behavior, salivation, and ruminal pH of lactating dairy cows. AB - Four multiparous and four primiparous lactating dairy cows fitted with ruminal cannulas were used in a duplicated 4 x 4 Latin square design to study the effects of parity and inclusion of a fibrolytic enzyme product (Agribrands International, St. Louis, MO) on feeding and chewing behavior, salivation, and ruminal pH. Diets consisting of rolled barley, barley silage, and alfalfa haylage (55% forage, DM basis) differed in enzyme application: 1) control, 2) enzyme applied to concentrate (45% of TMR), 3) enzyme applied to supplement (4% of TMR), and enzyme applied to a premix (0.2% of TMR). Enzyme supplementation did not alter daily time spent eating or ruminating, but when enzymes were added to the ration daily, saliva production increased, with no difference among enzyme application treatments. Multiparous cows consumed a greater amount of feed, but spent a similar amount of time eating, compared to primiparous cows. Primiparous cows had shorter ruminating episodes, resulting in lower daily ruminating time compared with multiparous cows. Primiparous cows had lower daily saliva output compared with multiparous cows. These results indicate that application of this fibrolytic enzyme product did not alter the physical structure of the feed, as measured by feeding and chewing variables. The increase in total saliva production observed in cows fed enzyme-supplemented diets may be attributed to a physiological response to compensate for the increase in fermentation products during digestion. The increased intake for multiparous cows is attributed to increased eating rate and not to increased time spent eating. The higher DMI of multiparous cows resulted in increased rumination time needed to process the additional feed and increased salivation to buffer the greater production of VFA. PMID- 12647964 TI - Effect of feeding supplemental fibrolytic enzymes or soluble sugars with malic acid on milk production. AB - Two trials were conducted to evaluate effects of feeding supplemental fibrolytic enzymes or soluble sugars and malic acid on milk production. In trial 1, 257 cows at four sites were fed a basal diet consisting of no more than 60% of forage DM as corn silage and less than 40% as alfalfa hay. Cows were assigned randomly within site, parity, and two stages of lactation to: 1) control; 2) enzyme A; 3) enzyme B; and 4) soluble sugars and malic acid. There was a 14-d pretreatment and an 84-d treatment period. Enzyme solutions were sprayed on either the forage component or the TMR each day while mixing feed. Trial 2 was similar, except 122 cows at one site in the United Kingdom were fed diets containing forage that was 75% corn silage and 25% grass silage, and all cows began the study between 25 to 31 DIM. Mean milk productions for 233 cows that completed trial 1 were 32.9, 32.5, 32.4, and 32.9 kg/d for control, enzyme A, enzyme B, and soluble sugars and malic acid, respectively. Mean milk productions for 116 cows that completed trial 2 were 28.2, 27.9, 28.8, and 28.4 kg/d, respectively. In vitro analyses of the activities of enzyme solutions indicated that all major cellulose and hemicellulose degrading activities were present; however, the pH optima (approximate pH = 4 to 5) were more acidic, and the temperature optimum (approximately 50 degrees C) was greater than normal pH and temperature in the rumen. If fibrolytic activity in the rumen is a major mechanism of action of supplemental fibrolytic enzymes, it appears that considerable activity of these preparations was lost due to conditions in the rumen. In conclusion, feeding supplemental fibrolytic enzymes or malic acid with soluble sugars had no effect on milk production under the conditions used in this study. PMID- 12647965 TI - Effects of spray-dried animal plasma in calf milk replacer on health and growth of dairy calves. AB - Male Holstein calves (n = 120) purchased from local dairy farms were fed one of three calf milk replacers for 42 d. Experimental milk replacers were formulated to contain whey protein concentrate (WPC) as the primary protein source or WPC plus 5% spray-dried bovine plasma (SDBP) or spray-dried porcine plasma (SDPP). The SDPP was heated to remove heat-insoluble materials and provide products with similar IgG content. Calves were also fed commercial calf starter and water for ad libitum consumption. Intake, change in body weight (BW), feed efficiency, morbidity and mortality were determined. Mortality was 10, 3, and 2 in calves fed WPC, SDBP, and SDPP treatments, respectively. Morbidity, measured as the number of days that calves had diarrhea was reduced by 30% when SDBP or SDPP were fed. Calves had diarrhea for 6.9, 3.9, and 4.7 d during the 42-d study when fed commercial calf milk replacer containing WPC, SDBP, and SDPP, respectively. Fecal scores tended to be reduced and feed efficiency tended to be improved when SDBP or SDPP were fed. Mean intakes of total dry matter during the 42-d study were greater when calves were fed SDBP or SDPP and were 661, 710, and 684 g/d for calves fed WPC, SDBP, and SDPP, respectively. Mean BW gains from d0 to 42 were 231,261, and 218 g/d, respectively. Calves fed SDPP tended to have lower BW gain during the first 28 d of the study. However, difference in daily BW gain from d 1 to 28 was only 39 g/d. Inclusion of SDBP or SDPP in milk replacer reduced morbidity and mortality of milk-fed dairy calves. PMID- 12647966 TI - Effects of tallow in diets based on corn silage or alfalfa silage on digestion and nutrient use by lactating dairy cows. AB - Six multiparous Holstein cows (average 31 days in milk; 36.3 kg/d of milk) fitted with ruminal cannulas were used in a 6 x 6 Latin square with 21-d periods to investigate the effects of diets that varied in forage source and amount of supplemental tallow. Isonitrogenous diets in a 2 x 3 factorial arrangement were based on either high corn silage (40:10 corn silage to alfalfa silage, % of dry matter) or high alfalfa silage (10:40 corn silage to alfalfa silage, % of dry matter) and contained 0, 2, or 4% tallow. Intakes of dry matter and total fatty acids were lower when cows were fed the high corn silage diet. Tallow supplementation linearly decreased dry matter intake. Milk yield was unaffected by diet; yields of milk fat and 3.5% fat-corrected milk were higher for the high alfalfa silage diet but were unaffected by tallow. Milk fat percentage was higher for the high alfalfa silage and tended to decrease when tallow was added to the high corn silage diet. Contents of trans-C18:1 isomers in milk fat were increased by high corn silage and tallow, and tended to be increased more when tallow was fed in the high corn silage diet. Ruminal pH and acetate:propionate were lower when high corn silage was fed. Ruminal acetate:propionate decreased linearly as tallow increased; the molar proportion of acetate was decreased more when tallow was added to the high corn silage diet. Ruminal liquid dilution rates were higher for the alfalfa silage diet; ruminal volume and solid passage rates were similar among diets. Total tract apparent digestibilities of dry matter, organic matter, crude protein, starch, energy, and total fatty acids were unaffected by diet. Digestibilities of neutral detergent fiber, acid detergent fiber, hemicellulose, and cellulose were lower when high corn silage was fed. The high alfalfa silage diet increased intakes of metabolizable energy and N, and increased milk energy and productive N. Tallow decreased the amount of N absorbed but had few other effects on utilization of energy or N. Tallow linearly increased concentrations of nonesterified fatty acids and cholesterol in plasma; cholesterol was increased by high alfalfa silage. Overall, forage source had more pronounced effects on production and metabolism than did tallow supplementation. Few interactions between forage source and tallow supplementation were detected except that ruminal fermentation and milk fat content were affected more negatively when tallow was fed in the high corn silage diet. PMID- 12647967 TI - Effect of genetic merit and concentrate supplementation on grass intake and milk production with Holstein Friesian dairy cows. AB - A total of 48 high genetic merit (HM) and 48 medium merit (MM) cows, each given a low (LC), medium (MC), or high (HC) level of concentrate supplementation, were used in a split-plot design experiment, which was run in three consecutive years, to evaluate animal production responses. Individual cow intakes were estimated twice each year while at pasture; measurement period 1 (MP1) was in May/June, and measurement period 2 (MP2) was in early September, corresponding on average to d 110 and 200 of lactation, respectively. In MP1, cows were offered 0 (LC), 3 (MC), and 6 kg (HC), whereas in MP2 the levels were 0 (LC), 0 (MC), and 4 kg (HC) of concentrate daily. Genotype had a significant effect on all milk production parameters in MP1 and MP2. The HM cows had the highest yield of milk, fat, protein, and lactose, whereas the MM cows had the highest milk fat, protein, and lactose concentrations. The HM cows had significantly higher grass dry matter intake (GDMI) estimates. In MP1, the average responses, per kg concentrate dry matter, was +1.10 kg of milk, +0.038 kg of protein, +0.032 kg of fat. The corresponding values in MP2 were +0.94 kg of milk, +0.037 kg of protein, and +0.025 kg of fat. The response to concentrate was linear and independent of preexperimental milk yield. In MP1, the partial regression coefficients relating daily GDMI to an increase in 1 kg of preexperimental milk yield (PMY), preexperimental BW (PBW), and concentrate intake (CI) were 0.123, 0.006, and 0.54, respectively, whereas the corresponding values in MP2 were 0.190,0.007, and 0.444, respectively. This study indicates that with high yielding dairy cows, on gras only GDMI of 17 kg of supporting milk yield of 30-kg/d is achievable. In this scenario, concentrate supplementation will result in lower substitution rates, and higher milk yield response than previously published with lower yielding cows. PMID- 12647968 TI - Effect of dietary ascorbyl-2-phosphate on immune function after transport to a feeding facility. AB - Effects of dietary ascorbyl-2-phosphate on immune function after a 210-km trip were measured in 18 Holstein heifers. After transport on d 0, 10 g of ascorbyl-2 phosphate each were added to the diets of 10 heifers, whereas eight heifers were fed a control diet. Plasma cortisol concentrations increased by an average of 25.6 microgram/ml on d 0 following transport, but by d 7 after transport had decreased to pretransport levels. Average daily gain was lower in heifers fed ascorbyl-2-phosphate from d 28 to 49 d after transport, but did not differ over the entire study. Feeding ascorbyl-2-phosphate maintained plasma ascorbate concentrations on d 7 post-transport, which decreased in control heifers. Plasma keyhole limpet hemocyanin antibody titers were significantly higher in control heifers from d 7 to 49. Mononuclear leukocyte proliferation responses were decreased on d 0 in lymphocytes stimulated by mitogens, with pokeweed mitogen stimulated cells showing less of a response than cells stimulated by the other mitogens. In the absence of mitogens, dietary ascorbyl-2-phosphate increased basal 3H-methyl thymidine incorporation by cultured lymphocytes. Across diets and mitogens, lymphocytes treated with cortisol showed decreased 3H-methyl thymidine incorporation. Transportation acted as a stressor, as evidenced by the increased plasma cortisol levels at d 0 immediately after transport, but immunological effects were not apparent by d 7. Feeding ascorbyl-2-phosphate maintained plasma ascorbate concentrations on d 7, but had negative effects on immune responses posttransport. PMID- 12647970 TI - Effects of abomasal infusion of conjugated linoleic acid on milk fat concentration and yield from pasture-fed dairy cows. AB - The aim of this study was to investigate the effects of conjugated linoleic acid supplementation on the synthesis of milk fat in pasture-fed Friesian cows. In four cows, a commercial mixture containing 62.3% (wt/vol) conjugated linoleic acid was infused intraabomasally to avoid rumen fermentation and biohydrogenation. The design was a 4 x 4 Latin square in which each cow received infusions of 0, 20, 40, and 80 g/d of conjugated linoleic acid mixture for 4 d. Cows were fed freshly cut ryegrass/white clover pasture ad libitum. Milk fat concentration was decreased by 36, 43, and 62% and milk fat yield was decreased by 32, 36, and 60% by the 20, 40, and 80 g of conjugated linoleic acid/d treatments. Dry matter intake, milk protein concentration, and protein yield were unaffected by treatments; however, milk yield was increased by 11% during the 40 g conjugated linoleic acid/d treatment. The effects of conjugated linoleic acid infusion were most pronounced in reducing de novo fatty acid synthesis and desaturation. Results show that the inhibitory effect of this conjugated linoleic acid mixture on milk fat synthesis occurs in pasture-fed cows, and demonstrate the potential to dramatically alter gross milk composition. This technology could offer a management tool to manipulate milk composition and energy demands of pasture-fed cows. PMID- 12647969 TI - Effects of particle size of alfalfa-based dairy cow diets on chewing activity, ruminal fermentation, and milk production. AB - Effects offorage particle size measured as physically effective NDF and ratio of alfalfa silage to alfalfa hay of diets on feed intake, chewing activity, particle size reduction, salivary secretion, ruminal fermentation, and milk production of dairy cows were evaluated using a 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments. The diets consisted of 60% barley-based concentrate and 40% forage, comprised either of 50:50 or 25:75 of alfalfa silage:alfalfa hay, and alfalfa hay was either chopped or ground. Various methods were used to determine physically effective NDF content of the diets. Cows surgically fitted with ruminal and duodenal cannulas were offered ad libitum access to these total mixed diets. The physically effective NDF content of the diets was significantly lower when measured using the Penn State Particle Separator than when measured based on particles retained on 1.18-mm screen. Intake of DM was increased by increasing the ratio of silage to hay but was not affected by physically effective NDF content of diets. Eating time (hours per day) was not affected by the physically effective NDF content of diets, although cows spent more time eating per unit of DM or NDF when consuming high versus low alfalfa hay diets. Ruminating time (hours per day) was increased with increased physically effective NDF content of the diets. Rumen pH was affected more by changing dietary particle size than altering the ratio of silage to hay. Feeding chopped hay instead of ground hay improved ruminal pH status: time during which ruminal pH was above 6.2 increased and time during which ruminal pH was below 5.8 decreased. Milk production was increased by feeding higher concentrations of alfalfa silage due to increased DM intake, but was not affected by dietary particle size. Feed particle size, expressed as mean particle length or physically effective NDF was moderately correlated with ruminating time but not with eating time. Although physically effective NDF and chewing time were not correlated to mean rumen pH, they were negatively correlated to the area between the curve and pH 5.8, indicating a positive effect on reducing the risk of acidosis. Milk fat content was correlated to rumen pH but not to physically effective NDF or chewing activity. These results indicate that increasing physically effective NDF content of the diets increased chewing activity and improved rumen pH status but had limited effect on milk production and milk fat content. PMID- 12647971 TI - Bivariate analysis of liability to clinical mastitis and to culling in first lactation cows. AB - Records taken on 13,070 first-lactation daughters of 250 Norwegian Cattle sires were used to examine associations between susceptibility to clinical mastitis and to culling. Clinical mastitis was defined as a binary trait, whereas culling was treated as either binary (culled or not culled) or continuous (length of opportunity period) for two sampling periods (120 or 300 d of lactation). Two Bayesian models were employed; 1) a bivariate threshold model with both mastitis and culling as binary traits, and 2) a bivariate model with mastitis as a threshold binary variable and time to culling as Gaussian. The heritability of liability to clinical mastitis was not affected by either the length of sampling period (120 vs. 300 d) or by whether culling (binary) or length of opportunity period was the second trait in the bivariate analysis. The posterior mean (standard deviation) of heritability of liability to clinical mastitis was 0.06 to 0.07 (0.02) in all analyses. The heritability estimate of length of opportunity period was less than 0.001. Culling (threshold trait) in first lactation had a low heritability, but a high genetic correlation with clinical mastitis. The posterior means (standard deviation) for heritability of liability to culling were 0.01 (0.006) for 120 d and 0.02 (0.009) for 300 d, and the posterior means (standard deviation) of the genetic correlation between liability to clinical mastitis and to culling were 0.48 (0.24) and 0.53 (0.21) for 120 and 300 d, respectively. PMID- 12647972 TI - Genetic parameters for feet and leg traits evaluated in different environments. AB - The objective of this study was to test for genotype x environmental interaction (GXE) for feet and leg traits scored in different environments. Genetic correlations of seven feet and leg traits were estimated across different management systems: free versus tie stalls, slatted versus solid flooring, and intact versus trimmed hooves. Data were records from first-lactation Holstein cattle. Traits were claw uniformity, depth of heel, rear leg rear view, foot angle, bone quality, rear leg side view, and overall feet and legs. Different subsets of data were used for each comparison, resulting in 147,400; 53,550; and 145,160 records for housing, flooring, and hoof trimming management systems, respectively. Genetic parameters were estimated using REML and two-trait models in which for each animal a given trait was observed in one environment and missing in the other. Phenotypic scores were lower with tie stalls, slatted floors, and no trimming. Heritabilities tended to be greater in herds with tie stalls and slatted floors. Trimming had little effect on genetic parameters. The genetic correlations of feet and leg traits across pairs of management systems were > or = 0.85, except for rear legs, rear view. Therefore, effects of GXE were assumed to be of little importance and modification of genetic evaluation procedures on the basis of housing, flooring, and hoof conditions seems unnecessary. PMID- 12647973 TI - Effects of including a quantitative trait locus in selection under different waiting plans of young bulls. AB - The advantage of using the genotype of a quantitative trait locus (QTL) in selection schemes of dairy cattle was quantified using stochastic simulation. Three selection plans were studied. In the first plan, young bulls waited for 3 yr until their sisters completed a lactation and then were evaluated and selected based on an animal model. In a second plan, young bulls waited for 5 yr until their daughters completed a lactation. An intermediate 4-yr waiting plan was also studied. Simulation was for 16 yr with overlapping generations. Population and model parameters were proportional to the U.S. Holstein population. The advantage of using a QTL was quantified as the percentage of superiority of QTL-assisted over QTL-free selection using cumulative genetic response. Percentage of superiority was reported for four selection pathways: active sires, young bulls, bull dams, and first lactation cows. A general trend was observed: low superiority in early years of selection that increased to a plateau in later years and then decreased. The superiority of the QTL information was greatest in the 3-yr waiting plan and least in the 4-yr waiting plan. Superiority at plateau for selection pathways ranged from 16 to 26% for the 3-yr waiting plan, from 3 to 12% for the 4-yr waiting plan, and from 5 to 13% for the 5-yr waiting plan. The contribution to selection response attributed to the QTL and the polygenes was quantified. The rate at which the favorable allele approached fixation and the accuracy of predicting breeding values on the percentage of superiority were studied. PMID- 12647974 TI - A simple method for weighted bending of genetic (co)variance matrices. AB - Due to computational demand, elements of genetic correlation matrices may have been estimated separately and then combined together into a single correlation matrix at a later stage. Because these matrices should be positive definite (PD) a statistical method commonly known as "bending" is used to make them positive definite. The conventional bending method ignores the reliability of different correlations and may subject any of them to change in order to make a positive definite correlation matrix. A simple method to obtain a weighted bended matrix to be used in animal breeding applications is proposed, and the implementation of the method is demonstrated by an example. PMID- 12647975 TI - Effect of omission of machine stripping on milk production and parlor throughput in East Friesian dairy ewes. AB - Due to the large cisternal storage capacity and non-vertical teat placement in most dairy ewes, machine stripping is commonly performed to remove milk not obtained by the machine. However, stripping requires individual manual intervention, lengthens the milking routine, and could inadvertently lead to overmilking of other ewes in the parlor. The objective of the present experiment was to estimate the effect of omission of machine stripping on milk production and parlor throughput. East Friesian crossbred dairy ewes that had been machine milked and stripped twice daily from d 0 to 79 postpartum, were randomly assigned to two stripping treatments for the remainder of lactation: normal stripping (S, n = 24), or no stripping (NS, n = 24). The NS ewes yielded 14% less commercial milk during the experiment, but had similar lactation length, milk composition, and somatic cell count compared to S ewes. Average machine milk yield (amount of milk obtained without manual intervention) tended to be greater for NS compared to S ewes. Average machine-on time for S ewes was longer than for NS ewes because of stripping, which may have resulted in over-milking of many ewes in the S group. Results from a milking simulation indicated that parlor throughput would increase by 33%, and overmilking would not occur when stripping was omitted from the milking routine. These results collectively suggest that residual milk left in the udder as a result of omission of machine stripping does not negatively influence milk quality and the loss in commercial milk yield could be compensated for by improved parlor throughput. PMID- 12647976 TI - Whole-farm phosphorus balance on western dairy farms. AB - Environmental concerns have focused attention on animal agriculture and its contribution to P accumulation in soils and runoff to surface waters. Monitoring P inputs and outputs on farms is a means of calculating the potential P build-up in farm soils. The objective of this study was to determine whole-farm P balance and the relative importance of the farm components (herd, manure storage, cropping systems) that contribute to it in dairies of the western United States. Whole-farm balances were computed for 41 commercial dairies in Utah and Idaho using the Maryland Nutrient Balancer. The average whole-farm P balance in the study was 6.6 tonne/yr with an average herd size of 466 cows. Imported feed made up 85.4% of the total P inputs and exported animal products (milk and meat), and manure and compost made up 53.1 and 45.9%, respectively, of the total P outputs. Farms were divided into those that grew crops and those that did not. Whole-farm balance (kg of P balance per animal) for farms that grew crops had more unaccounted for P (difference between P inputs and output) than farms that grew no crops. They also had more imported fertilizer and less imported feed and exported manure and compost. Multiple regression analysis of the relative effects of herd management, manure storage, and cropping system on whole-farm balance per product found that herd P utilization efficiency was the most important factor in determining whole-farm P balance on farms where crops were grown. Crop uptake of available P was the only other subsystem important for these farms. Increased conversion of feed P to P in product is an important way to decrease whole-farm P balance. PMID- 12647977 TI - American society of clinical oncology 2002. Orlando, Florida, USA, 18-22 May, 2002. PMID- 12647978 TI - 5th International Congress of Nuclear Oncology and 15th National Congress of Turkish Society of Nuclear Medicine Kusadasi, Turkey, 1-5 May, 2002. PMID- 12647980 TI - Darbepoetin alfa: potential role in managing anemia in cancer patients. AB - Anemia in cancer patients is frequent but often under-recognized and under treated. This may be related to misconceptions about the impact of anemia on cancer patients and ill-defined guidelines for treatment, as well as the inadequacies of current therapy. Darbepoetin alfa, a novel erythropoiesis stimulating protein with a longer serum half-life than epoetin (alpha and beta), is approved to treat anemia in patients with chronic kidney disease. Most recently darbepoetin alfa has received approval by the FDA in USA for the treatment of anemia associated with myelosuppressive chemotherapy and approval in the EU is expected soon. Clinical trials in cancer patients indicate that darbepoetin alfa effectively and safely alleviates anemia in patients receiving chemotherapy. A Phase II trial also indicates that darbepoetin alfa is effective in patients who are not receiving chemotherapy. Thus, darbepoetin alfa has the potential to improve supportive care and thereby, cancer patients' quality of life, and might also impact on treatment outcome. PMID- 12647979 TI - Management of symptoms associated with advanced cancer: olanzapine and mirtazapine. A World Health Organization project. AB - Advanced cancer patients are polysymptomatic and often receive multiple medications for symptom relief. Common symptoms include anorexia, weight loss, delirium and depression. Olanzapine and mirtazapine may have several advantages over older agents despite increased acquisition costs. Both medications can treat several symptoms with a low risk for drug-drug interactions and with only once- or twice-daily dosing. Drug side effects are low, compared with more conventionally used agents. The pharmacokinetics and pharmacodynamics of both agents are unique and explain many of the benefits. More research and clinical experience will be necessary to define their role in the palliation of advanced cancer. PMID- 12647981 TI - Radiologic screening for lung cancer. AB - Lung cancer is the leading cause of cancer death. A number of screening trials have been carried out over the past two decades and have failed to demonstrate a reduction of lung cancer-related mortality. Recent advances in new forms of technology--spiral low-dose CT--have resuscitated interest in screening programs. Preliminary results are promising but the outstanding question is whether the magnitude of improvement is sufficient to affect a measurable reduction in lung cancer mortality? PMID- 12647982 TI - Advanced NSCLC: from cytotoxic systemic chemotherapy to molecularly targeted therapy. AB - Approximately a third of non-small cell lung cancer patients present with disseminated disease at the time of diagnosis. For these patients, as well as those with recurrent disease, chemotherapy remains the mainstay of treatment. For several decades, researchers have attempted different combinations of drugs in search for the 'best' chemotherapy regimen. Despite the emergence of newer, 'third-generation' cytotoxic agents, success is still modest at best. Fortunately, new insights in tumor biology, leading to the design of molecularly targeted drugs, are opening a new era in cancer treatment. These novel agents target molecular pathways specifically found in cancer cells, thus maximizing the antitumor effect while minimizing toxicities on normal cells. PMID- 12647983 TI - Chemoradiotherapy of non-small cell lung cancer. AB - The efficacy of radiotherapy in locally advanced unresectable non-small cell lung cancer is low. This method does not provide effective eradication of bulky disease in the thorax, neither does it prevent uncontrolled systemic disease. The addition of chemotherapy to radiation results in increased cure rate by both improving tumor control in the thorax and by eliminating or delaying the emergence of metastatic disease. The two most frequently tested strategies of combining chemotherapy and radiation include primary chemotherapy followed by radiation and concurrent application of both methods. This review provides the rationale for this strategy and presents the results of major Phase III studies. Discussed are advantages of chemoradiation, its limitations in clinical practice and prospects for the future. PMID- 12647984 TI - Fluoropyrimidine resistance in colon cancer. AB - A significant obstacle for the management of patients with colorectal cancer is intrinsic drug resistance or in patients that respond to chemotherapy, acquired drug resistance. Drug resistance can occur through a variety of mechanisms. These include alterations in drug influx, drug efflux, intracellular metabolic activation, intracellular catabolism, through alterations in the drug's target or through numerous changes downstream of the target including alterations in genes involved in the regulation of the cell cycle, apoptosis or in DNA damage repair. In this article, the mechanisms of action and the mechanisms of resistance to the fluoropyrimidines are reviewed focusing on newer studies using tumor samples obtained from patients. Clinical trials that can potentially overcome the relevant mechanisms of resistance are described. PMID- 12647985 TI - Contribution of meta-analyses to the evaluation of treatments for advanced colorectal cancer. AB - In the last decade, the Meta-Analysis Group in Cancer (formerly Advanced Colorectal Cancer Meta-Analysis Project) has systematically used a meta-analytic to reassess the efficacy and toxicity of various fluoropyrimidine regimens in advanced colorectal cancer, as well as to investigate related questions, such as the relation between tumor response and survival. In this paper, the basic methodology of meta-analysis is reviewed and the findings of seven successive meta-analyses conducted between 1989-2000 by the Meta-Analysis Group in Cancer are summarized. The interested reader can refer to the statistical appendix for details on the methodology used for all meta-analyses. PMID- 12647987 TI - Evolving role of ribonucleoside reductase inhibitors in hematologic malignancies. AB - Ribonucleotide reductases catalyze the de novo biosynthesis of deoxyribonucleosides for DNA synthesis. Increased ribonucleotide reductases activity has been associated with malignant transformation and tumor cell growth. The ribonucleotide reductases inhibitors may bind with the R1 subunit of the enzyme (Class 1) or the nonheme iron (Class 2). This review focuses on the therapeutic use of ribonucleotide reductases inhibitors in hematologic malignancies. Hydroxyurea, fludarabine and cladribine have established roles in the management of hematologic malignancies, while other ribonucleotide reductases inhibitors, such as gemcitabine, tezacitabine and heterocyclic carboxaldehyde thiosemicarbazones (e.g., triapine) are being evaluated in clinical trials. PMID- 12647986 TI - Systemic therapy for advanced pancreatic cancer. AB - Death from pancreatic cancer remains high with few long-term survivors. Systemic chemotherapy with 5-fluorouracil-based combinations had minimal impact on natural history of this disease. Several new agents with activity against pancreatic cancer have been identified over the past decade. Gemcitabine has modest activity in this disease. Combination chemotherapy trials incorporating gemcitabine, cisplatin, 5-fluorouracil, oxaliplatin, docetaxel or irinotecan show improved outcomes in objective response rates and survival that need to be confirmed in prospectively randomized studies. Advancement in the understanding of the biology of pancreatic cancer has helped identify several molecular targets for the development of novel therapies. Ongoing and future treatment regimens for pancreatic cancer will incorporate traditional cytotoxic drugs and novel targeted therapies. PMID- 12647988 TI - Integrins and extracellular matrix: a novel mechanism of multidrug resistance. AB - Multidrug resistance remains the major hurdle to successful cancer treatment. Classical mechanisms of multidrug resistance include drug efflux pumps, glutathione-S-transferase upregulation and topoisomerase II-associated multidrug resistance. However, despite extensive research, the clinical relevance of these mechanisms remains unclear and no significant clinical benefit has materialized. Recently, a novel mechanism of multidrug resistance has been identified- extracellular matrix-mediated multidrug resistance: integrin-mediated adherence of cells to extracellular matrix proteins results in significant resistance to many anticancer agents that induce cell death via unrelated mechanisms. Verification of the mechanisms of action of this novel phenomenon will hopefully identify new therapeutic targets to aid in the fight against cancer. PMID- 12647989 TI - Interference with EGFR signaling: paradigm for improving radiation response in cancer treatment. AB - The introduction of biologically active agents that interfere with the epidermal growth factor receptor (EGFR) provides a promising opportunity to improve cancer treatment outcomes. Several EGFR-selective agents, such as humanized monoclonal antibodies and small molecule, orally available tyrosine kinase inhibitors have shown antitumor activity in early clinical trials in advanced cancer patients. Preclinical studies have demonstrated enhanced radiation- and chemotherapy induced tumor cytotoxicity when EGFR antagonists are implemented. More broadly, recent clinical trials have confirmed improved survival with combinations of HER 2 (a member of the ErbB family of receptors) targeted antibodies and chemotherapy in patients with advanced breast cancer. A landmark trial combining C225 antiEGFR antibody with radiation therapy for patients with locally advanced head and neck cancer has just completed accrual. Thus, emerging rapidly are cancer treatment strategies based on an improved understanding of the specific cellular and molecular abnormalities of individual tumors. PMID- 12647990 TI - DNA diagnostics in the fifty-year retrospect. PMID- 12647991 TI - Molecular diagnostics: an FDA perspective. AB - The potential medical applications of microarrays and in vitro diagnostic devices for global assessments of DNA sequence variations, relative RNA abundance and measurements of proteins have generated much excitement, and some skepticism, within the biomedical community. It has been suggested that within the next decade these microarrays and diagnostic devices will be routinely used in the selection, assessment and quality control of the best drugs for pharmaceutical development, at the bedside for diagnostics and for clinical monitoring of both desired and adverse outcomes of therapeutic interventions. Realizing such potential will be a challenge to the entire scientific community as often breakthroughs which show great promise at the bench fail to meet the requirements of clinicians and regulatory scientists, and to make the transition into common clinical and regulatory practice. The development of a co-operative framework between regulators, product sponsors and technology experts will be essential for realizing the revolutionary promise these platforms could have on the evolution of drug development, regulatory science, the practice of medicine and public health. PMID- 12647992 TI - Origin and utility of the reverse dot-blot. AB - Reverse allele specific oligonucleotide assays provide a robust method for the molecular characterization of high-mutation spectrum disorders. Commercial test have been developed for human leukocyte antigens class I and class II regions of human chromosome 6, the cystic fibrosis transmembrane conductance regulator at 7q31 and strains of human Hepatitis B and C virus. In their most developed form, these assays rely upon highly multiplexed PCR reactions containing biotinylated primers providing a substrate for nonradioactive detection systems. Sophisticated reverse dot-blot technology involves mechanized covalent attachment of activated primary amine-conjugated oligonucleotides to carboxylated nylon membranes or bovine serum albumin. Subsequent to line or dot printing, membranes are stored or sold dry in preparation for hybridization. Circular spots or lines are visualized colorimetrically after hybridization through the use of streptavidin horseradish peroxidase incubation followed by development using tetramethylbenzidine and hydrogen peroxide, or via chemiluminescence after incubation with avidin alkaline phosphatase conjugate and a luminous substrate susceptible to enzyme activation, such as CSPD, followed by exposure to x-ray film. The entire procedure from blood specimen receipt to result usually requires less than 1 day. Because of the simplicity, speed, and generally high sensitivity and specificity, large numbers of individuals can be rapidly screened using this technology. Rapid turnaround is often required in prenatal diagnosis of cystic fibrosis, beta-thalassemia and hemoglobinopathies, giving this technology has special applicability in those genetic diseases. Commercial instruments are available which automate the hybridization and color development. In addition, scanning software can capture the probe reactivity pattern and interpret it in terms of a genotype. PMID- 12647993 TI - Nanodiagnostics: application of nanotechnology in molecular diagnostics. AB - Nanotechnology extends the limits of molecular diagnostics to the nanoscale. Nanotechnology-on-a-chip is one more dimension of microfluidic/lab-on-a-chip technology. Biological tests measuring the presence or activity of selected substances become quicker, more sensitive and more flexible when certain nanoscale particles are put to work as tags or labels. Magnetic nanoparticles, bound to a suitable antibody, are used to label specific molecules, structures or microorganisms. Magnetic immunoassay techniques have been developed in which the magnetic field generated by the magnetically labeled targets is detected directly with a sensitive magnetometer. Gold nanoparticles tagged with short segments of DNA can be used for detection of genetic sequence in a sample. Multicolor optical coding for biological assays has been achieved by embedding different-sized quantum dots into polymeric microbeads. Nanopore technology for analysis of nucleic acids converts strings of nucleotides directly into electronic signatures. DNA nanomachines can function as biomolecular detectors for homogeneous assays. Nanobarcodes, submicrometer metallic barcodes with striping patterns prepared by sequential electrochemical depositon of metal, show differential reflectivity of adjacent stripes enabling identification of the striping patterns by conventional light microscopy. All this has applications in population diagnostics and in point-of-care hand-held devices. PMID- 12647994 TI - Biomarkers in melanoma: predisposition, screening and diagnosis. AB - Melanoma's incidence and mortality for certain groups has appeared to level off after a steady increase over the last half century. This trend is suspected to be due to better detection and removal of thin, biological early melanomas. However, to date, no prospective evidence exists to clearly demonstrate the efficacy of prevention and early detection in decreasing melanoma mortlity. Nonetheless, many studies suggest that both self-assessment of risk factors or clinician examination can identify a proportion of patients at highest risk for melanoma who may benefit from behavior modification (primary prevention) and routine screening (secondary prevention). Compromising these goals is the fact that neither the clinical or histologic diagnosis of melanoma is 100% accurate. Clinical diagnosis of melanoma, based on evaluation of a skin lesion's color and shape, correlates best with the experience of the clinician. Ancillary technologies have been developed to improve clinical accuracy of suspicious skin lesions but a subset of melanomas exist that do not fall in the spectrum of the 'ABCDE' guidelines commonly used for melanoma identification. Similarly, at the histologic level (the 'gold standard' of diagnosis), overlap exists between benign and malignant melanocytic proliferations leading to both over and underdiagnosis of melanoma. A better understanding of melanoma's pathogenesis has identified disease-related biomarkers that may more reliably differentiate a melanocytic nevus from melanoma. In this paper, we review current and novel, potentially more accurate, biomarkers and supplementary technologies that can be used for the prevention, screening and diagnosis of melanoma. PMID- 12647995 TI - Gene expression in cancer: the application of microarrays. AB - Genome-wide monitoring of gene expression using DNA microarrays represents one of the latest breakthroughs in experimental molecular biology and provides unprecedented opportunity to explore the biological processes underlying human diseases by providing a comprehensive survey of a cell's transcriptional landscape. In the cancer field, this revolutionary technology allows the simultaneous assessment of the transcription of tens of thousands of genes, and of their relative expression between normal cells and malignant cells. As microarray analysis emerges from its infancy, there is widespread hope that microarrays will significantly impact on our ability to explore the genetic changes associated with cancer etiology and development, and ultimately lead to the discovery of new biomarkers for disease diagnosis and prognosis prediction, and of new therapeutic tools. This review provides an overview of microarray technology, specifically in the context of cancer research and describes some of its recent applications to the study of cancer. In addition, the challenges of translating microarray findings into molecular cancer diagnosis and prognosis tools, with the potential of altering clinical practice through individualized cancer care and ultimately of contributing to the battle against cancer, are discussed. PMID- 12647996 TI - Genetic screening for hereditary diffuse gastric cancer. AB - Knowledge regarding the genetic basis of hereditary diffuse gastric cancer has greatly increased in the past 4 years, namely due to the discovery of segregating germline mutations in the gene coding for E-cadherin, within families with this cancer predisposing syndrome. Members of hereditary diffuse gastric cancer families have predominantly high predisposition to develop diffuse carcinomas of the stomach but can also be associated with an elevate risk for other types of cancer, namely lobular breast carcinoma. In this review, we focus on the epidemiology, pathology and genetics of gastric cancer, describe families, E cadherin mutations, and suggest alternative candidate genes underlying the hereditary diffuse gastric cancer syndrome. This knowledge is a fundamental step towards accurate genetic counselling, in which a highly specialized presymptomatic therapeutic intervention should be offered. PMID- 12647998 TI - Detection of target nucleic acids and proteins by amplification of circularizable probes. AB - Circularizable oligonucleotide probe (C-probe) is a unique molecule that offers significant advantages over conventional probes. Closed circular structure can be formed through ligation of its ends after hybridizing onto a target and locked on its target due to the helical turns formed between the complementary sequences of the target and the C-probe (padlock probe). Under an isothermal condition, C probe can be amplified by rolling circle amplification, to generate multimeric single-stranded DNA. This multimeric single-stranded DNA can be further amplified by a ramification mechanism through primer extension and upstream DNA displacement, resulting in an exponential amplification. Usually, an unbiased product is generated by either rolling circle amplification or ramification mechanism due to the generic primers of C-probe and is localized on targets. These advantages make C-probe amplification very useful for research and molecular diagnosis, especially in the areas where other techniques are not adequately helpful. The development of C-probe-based technologies iniates a new future for molecular diagnostics. The applications of C-probe, rolling circle amplification, ramification mechanism, in situ detection, microarray, immunoassay, single nucleotide polymorphism and whole genome amplification are discussed. PMID- 12647997 TI - Gene expression profiling of inflammatory bladder disorders. AB - Inflammation underlies all major bladder pathologies including malignancy and represents a defense reaction to injury caused by physical damage, chemical substances, micro-organisms or other agents. During acute inflammation, activation of specific molecular pathways leads to an increased expression of selected genes whose products attack the insult, but ultimately should protect the tissue from the noxious stimulus. However, once the stimulus ceases, gene expression should return to basal levels to avoid tissue damage, fibrosis, loss of function, and chronic inflammation. If this down-regulation does not occur, tissue fibrosis occurs as a serious complication of chronic inflammation. Although sensory nerve and most cells products are known to be key parts of the inflammatory puzzle, other key molecules are constantly being described that have a role in bladder inflammation. Therefore, as the database describing the repertoire of inflammatory mediators implicated in bladder inflammation increases, the central mechanisms by which injury can induce inflammation, cell damage, and repair often becomes less rather than more clear. To make sense of the vast knowledge of the genes involved in the inflammatory response may require analysis of the patterns of change and the elucidation of gene networks far more than definition of additional members of inflammatory cascades. This review discuss the appropriate use of microarray technology, which promises to solve both of these problems as well as identifying key molecules and mechanisms involved in the transition between acute and chronic inflammation. PMID- 12647999 TI - Rapid enhanced tissue culture immunofluorescence: a new, rapid method for detection of viruses. AB - The detection of a large variety of viruses requires a number of different susceptible cell lines in a shell vial method--currently considered to be the most enhanced method of viral detection. However, this method is unsuitable in its standard form for handling a large volume of specimens. We have developed a time and cost saving new method, named rapid enhanced tissue culture immunofluorescence (RETCIF) and incorporates the attributes of the current methods. It is specific, uses the most sensitive cell lines and monoclonal antibodies and does not require the use of cover slips. The RETCIF method, in our hands, is a time saving procedure, with higher isolation ratio than the shell vial method. We recommend the RETCIF method for busy diagnostic virology laboratories. PMID- 12648000 TI - A review of treatment for carpal tunnel syndrome. AB - PURPOSE: A common disorder encountered by healthcare specialists is carpal tunnel syndrome (CTS). CTS is a neuropathy disorder caused by compression on the median nerve. METHOD: Currently, there are several treatment methods for CTS such as: (1) non-steroidal anti-inflammatory drugs (NSAIDs); (2) injection of medications; (3) immobilization by splinting; (4) rehabilitation modalities (therapeutic ultrasound, ASTM AdvantEDGE, stretching and strengthening); and (5) surgery by carpal tunnel release. RESULTS: While NSAIDs, injections, and splinting have shown promise in relieving symptoms, long-term outcomes have been poor. CONCLUSION: This article provides a background in current treatment methods and an insight into the focal point of the future. PMID- 12648001 TI - A clinical model for the assessment of posture and balance in people with stroke. AB - PURPOSE: The lack of models to define and describe rehabilitation processes have often been identified as limiting research and the development of clinical practice. This study describes the development of a clinical model to address a key aspect of stroke physiotherapy--the assessment of posture and balance. METHOD: Twenty seven experienced neurological physiotherapists (PT) in six focus groups were used. Participants were shown photographs of a typical stroke patient in sitting and standing positions and were asked 'What would you note if you were assessing the posture and balance of this patient?' Answers were displayed on flip charts to allow immediate feedback about the accuracy and completeness of data. Thematic content analysis was then used. RESULTS: A complex reasoning process emerged to answer three main questions: What can the patient do? How does s/he do it? Why does s/he do it that way? To answer these questions physiotherapists established balance disability (by observing the patient's ability to perform a series of increasingly demanding balance tasks), identified postural and movement impairments (by observing alignment and movement of body segments relative to each other and to the expected norm for that patient) and assessed muscle activity (by observation and palpation). CONCLUSIONS: Focus groups have been used to elicit a clinical model for the assessment of posture and balance, the content of which will be used to inform a new outcome measure. PMID- 12648002 TI - A postural adaptation test for stroke patients. AB - PURPOSE: To explore the interrater reliability of the Advanced Mobility and Balance Scale (AMBS) and to determine its discriminative capacity in stroke patients. METHODS: Twelve hemiparetic patients and six healthy elderly volunteers were videotaped while: (1) executing rapid head motions during standing and walking; and (2) standing and walking on a slope. Five physical therapists viewed the videotapes to establish interrater reliability. RESULTS: Interrater reliability: Intraclass correlation ratios ranged from 0.93-0.97 for the AMBS global as well as slope and head turn subscores. Construct validity: One-way ANOVAs and post-hoc pairwise comparisons were performed to determine whether there was a difference in scores between high (HFL) and low functional-level (LFL) stroke patients (based on gait speed) and healthy subjects. Mean (+/-SD) global scores were 45 +/- 3 for healthy subjects, 40 +/- 9 for HFL stroke patients and 25 +/- 1 for LFL stroke patients (p < 0.05 for HFL versus LFL patients and LFL patients versus healthy subjects). The AMBS slope subscores were 22 +/- 2, 19 +/- 5, 9 +/- 7 for healthy, HFL and LFL subjects respectively (p < 0.05 for HFL versus LFL patients and LFL patients versus healthy subjects). CONCLUSION: The AMBS has excellent interrater reliability and good discriminative capacities. PMID- 12648003 TI - Physicians' preferences and expectations in traumatic brain injury rehabilitation -results of a case-based questionnaire survey. AB - PURPOSE: Analysis of medical decisions in the differential allocation of traumatic brain injury (TBI) rehabilitation programmes. METHOD: Medical specialists routinely involved in the treatment and rehabilitation of TBI patients from acute hospitals, rehabilitation centres, and the Medical Services of the Workers' Pension Insurance of Sachsen-Anhalt and Mecklenburg-Vorpommern were included in a systematic survey, in which they had to give their opinions on the individually optimal rehabilitation setting and therapies and probable outcome on the basis of case vignettes. These specialists are crucial for decision-making in rehabilitation resource allocation. RESULTS: Both allocation and prognosis depended mainly on patient characteristics. In a mixed logistic regression model, which takes into account that: (a) patient and physician characteristics may influence each other; and (b) that expert assessments might be correlated, setting preferences were related to chronicity and degree of motor and cognitive impairment, but also to the institutional background of the reviewer. Neuropsychological therapy dominated intervention recommendations, followed by physio- and occupational therapy. A comparatively great demand of psychotherapy and medicosocial assessment was recognized. Preferred rehabilitation setting did not influence the physicians' subjective prognosis. Older, chronic and motorically impaired patients were considered to profit less from rehabilitation. Physician characteristics and recommendations of either rehabilitation setting or therapies did not influence prognostic expectations. CONCLUSIONS: Physician characteristics influenced setting preferences. Preferences for type of treatment and ADL-prognosis depended on patients' variables only. Expected effects of rehabilitation were independent on rehabilitation setting and kinds of treatment. PMID- 12648004 TI - Is the Rivermead Mobility Index appropriate to measure mobility in lower limb amputees? AB - PURPOSE: Measurement of lower limb amputee mobility was investigated using the Rivermead Mobility Index (RMI). METHOD: Reliability and reproducibility were assessed in 62 patients. The timed waking test (TWT) was used to investigate concurrent validity. The RMI construct was examined in 200 established amputees. RESULTS: One hundred and forty-four males and 66 females, aged 13-90 were recruited. Intraclass correlation coefficients and kappa statistics showed good reproducibility. Spearman correlation coefficient between the RMI and TWT -0.58 (p<0.000). Psychometric properties of the RMI were tested using item response theory. Hierachical differences in RMI grades were identified in amputees compared with neurologically impaired patients for which the RMI was developed. The RMI construct was not unidimensional, with redundancy of items and local dependency. At the upper end of the scale there were insufficient items measuring high levels of mobility. Finally, differential item functioning showed items behaving differently for patient subgroups. CONCLUSIONS: Although initial impressions suggest the RMI is a useful measure of lower limb amputee mobility, further analysis shows it is not appropriate for all amputees, with a number of limitations of its psychometric properties. Its use is not recommended in this population. PMID- 12648005 TI - Classifying client goals in community-based ABI rehabilitation: a taxonomy for profiling service delivery and conceptualizing outcomes. AB - PURPOSE: To develop, confirm and trial a framework for analysing the content of goals set within community-based rehabilitation. This framework (taxonomy) is proposed as a tool to assist in service evaluation and outcome exploration. METHOD: Qualitative thematic analysis and categorization of 1765 rehabilitation goal statements in a four phase process of synthesis, refinement, verification and application. RESULTS: A taxonomy of goal content was developed comprising 21 categories within five domains, utilizing 125 descriptors. The taxonomy demonstrated good inter-rater consistency and was able to discriminate between similar but related data sets comprising goal statements. CONCLUSION: Structured analysis of the content of goal setting (particularly in community rehabilitation) utilizing a framework such as the proposed taxonomy has considerable potential as a 'window' into service delivery to broaden the parameters of existing service evaluation and to more clearly link outcome exploration to intervention. PMID- 12648006 TI - A head movement image (HMI)-controlled computer mouse for people with disabilities. AB - PURPOSE: This study proposes image processing and microprocessor technology for use in developing a head movement image (HMI)-controlled computer mouse system for the spinal cord injured (SCI). METHOD: The system controls the movement and direction of the mouse cursor by capturing head movement images using a marker installed on the user's headset. In the clinical trial, this new mouse system was compared with an infrared-controlled mouse system on various tasks with nine subjects with SCI. RESULTS: The results were favourable to the new mouse system. The differences between the new mouse system and the infrared-controlled mouse were reaching statistical significance in each of the test situations (p<0.05). CONCLUSIONS: The HMI-controlled computer mouse improves the input speed. People with disabilities need only wear the headset and move their heads to freely control the movement of the mouse cursor. PMID- 12648007 TI - Oral sustained delivery of paracetamol from in situ gelling xyloglucan formulations. AB - The purpose of this study was to evaluate the potential of a xyloglucan formulation with in situ gelling properties for the oral sustained delivery of paracetamol. Gelation of dilute aqueous solutions of the polysaccharide xyloglucan occurred in rabbit and rat stomachs as the orally administered chilled solutions attained body temperature. In vitro studies demonstrated diffusion controlled release of paracetamol from the gels over a period of 6 hr. The bioavailabilities of paracetamol from the xyloglucan gels formed in situ in the stomachs of rabbits after oral administration of the liquid formulations were similar to that of a commercially available suspension containing an identical dose of paracetamol. PMID- 12648009 TI - Melt granulation and heat treatment for wax matrix-controlled drug release. AB - The purpose of this study was to evaluate sustained drug release after melt granulation and heat treatment. Theophylline (anhydrous) and phenylpropanolamine hydrochloride (PPA) were used as model drugs. Compritol 888 ATO (Glyceryl Behenate NF) was incorporated as the wax matrix material. Formulations with drug:wax in 3:1 and 1:1 ratios were evaluated. Tablets were made by dry blending or melt granulation; some of the tablets were heat treated at 80 degrees C for 30 min. Tablets with or without heat treatment were tested for drug release using in vitro drug dissolution. The results showed that melt granulation gave slower drug release than dry blending. Heat treatment further retarded drug release for both dry blending and melt granulation. The drug release rates for theophylline were slower than for PPA at the same wax level and processing method. The drug release profiles were linear using a square root of time scale. In conclusion, melt granulation and heat treatment slowed drug release for the wax matrix-controlled release tablets. Heat treatment of the tablets made by melt granulation further retarded drug release. Heat treatment redistributed the wax, forming a new matrix system with higher tortuosity. The application of melt granulation or heat treatment can successfully retard drug release. PMID- 12648008 TI - Effect of buffers on the properties of microbicidal hydrogels containing monoglyceride as the active ingredient. AB - Hydrogel formulations containing the monoglyceride monocaprin have shown potent microbicidal activity against several sexually transmitted viruses and bacteria. It is recommended that formulations for preventing infection in the vagina have a low pH as the HIV virus is inactivated at low pH. The object of the work was to investigate how incorporation of buffers into the hydrogel formulations affects physicochemical properties and microbicidal activity of the active substance. Two series of gels were formulated using carbomer (Carbopol 934) and sodium carboxymethylcellulose (NaCMC) as gel-forming agents. The presence of buffers in the gels caused a lowering in gel viscosity, with carbomer gels being more sensitive to buffer presence than NaCMC gels. To obtain viscosity similar to that of a gel without buffer, the amount of polymer needs to be increased. An increase in the amount of NaCMC by 60-70% is needed to obtain the same viscosity as in gel without buffers; but for carbomer, the amount of polymer needs to be doubled. It appears that the effect of maleate buffer on NaCMC gel formation is greater than that of the citrate/lactate buffer; but for carbopol gels, the effects of the buffer systems tested on gel viscosity were equal. The virucidal activity of NaCMC gel buffered with citrate/lactate buffer against herpes simplex virus type 1 and HIV was not reduced by the presence of buffer. The results show that the presence of buffers in the hydrogel formulations affects gel viscosity, but the virucidal effect of the active compound, monocaprin, is not diminished. PMID- 12648010 TI - Carbamazepine/betaCD/HPMC solid dispersions. I. Influence of the spray-drying process and betaCD/HPMC on the drug dissolution profile. AB - The aim of this study was to compare carbamazepine (CBZ) solid dispersions prepared by spray-drying of aqueous dispersions with the corresponding physical mixtures. The influence of the association of beta-cyclodextrin (betaCD) and hydroxypropyl methylcellulose (HPMC) on the CBZ dissolution profile of the preparations was investigated. Results demonstrated that CBZ release from solid dispersions is dependent on the ratio of betaCD and HPMC. The spray-drying process confers better homogeneity to CBZ polymeric dispersions than the physical mixture process. In summary, we demonstrated the feasibility of obtaining a homogeneous polymeric solid dispersion of CBZ from an aqueous media by spray drying and a clear influence of the betaCD:HPMC ratio on the release profile of CBZ. PMID- 12648011 TI - Carbamazepine/betaCD/HPMC solid dispersions. II. Physical characterization. AB - Solid dispersions containing carbamazepine (CBZ) associated with beta cyclodextrin (betaCD) and/or hydroxypropyl methylcellulose were prepared by two different methods, spray-drying or physical mixture, and characterized by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), infrared (IR) spectroscopy, and x-ray powder diffraction analysis (XRPD) studies. Scanning electron microscopy pictures showed that spray-drying produced a mixture of hollow, spherical, and partially shrunken microparticles of homogeneous materials, whereas the physical mixtures yielded heterogeneous systems in which all individual components could be identified. Thermal and IR analyses suggest the existence of a strong interaction between CBZ and excipients in spray-dried solid dispersions, but no CBZ polymorphic transition was detected by either IR spectroscopy or XRPD analysis after the spray-drying process. PMID- 12648012 TI - Piroxicam benzoate--synthesis, HPLC determination, and hydrolysis. AB - An improved method of piroxicam benzoate synthesis was described, and an isocratic reversed-phase high-performance liquid chromatography method for its determination was developed and fully validated. The method was found to be specific, precise (relative standard deviation 0.3%), accurate (mean recovery 99.9%), and robust. Limit of detection was estimated at 0.055 microg mL(-1) and limit of quantification at 0.185 microg mL(-1). The kinetics of piroxicam benzoate hydrolysis in aqueous buffer solutions (pH 1.1 and 10), simulated gastric and intestinal fluids was studied. The hydrolysis followed first-order kinetics. The following rate constants were obtained at pH 10: k = 1.8 x 10(-3) hr(-1) at 37 degrees C and k = 3.4 x 10(-2) hr(-1) at 60 degrees C. In acidic media, no significant hydrolysis was observed after 24 hr. During the 24-hr period in simulated intestinal fluid, only 10.9% of the starting ester was hydrolyzed. PMID- 12648013 TI - N-trimethyl chitosan chloride: optimum degree of quaternization for drug absorption enhancement across epithelial cells. AB - N-trimethyl chitosan chloride (TMC) is a polycation that enhances drug transport across epithelia by opening tight junctions. The degree of quaternization of TMC determines the number of positive charges available on the molecule for interactions with the negatively charged sites on the epithelial membrane and thereby influences its drug absorption-enhancing properties. The effects of six different TMC polymers (degree of quarternization between 12% and 59%) on the transepithelial electrical resistance (TEER) of Caco-2 cell monolayers and on the transport of hydrophilic and macromolecular model compounds across Caco-2 cells were determined. All the TMC polymers were able to decrease the TEER markedly in a slightly acidic environment (pH 6.2). However, only TMC polymers with higher degrees of quaternization (> 22%) were able to reduce the TEER in a neutral environment (pH 7.4). The maximum reduction in TEER (47.34 +/- 6.0% at a concentration of 0.5% w/v and pH 7.4) was reached with TMC with a degree of quaternization of 48%, and this effect did not increase further with higher degrees of quaternization of TMC. In agreement with the TEER results, the transport of model compounds across Caco-2 cell monolayers increased with an increase in the degree of quaternization of TMC. However, the transport reached a maximum for TMC with a degree of quaternization of 48% (25.3% of the initial dose for [14C]mannitol and 15.2% of the initial dose for [14C]PEG 4000), and this effect did not increase further with higher degrees of quaternization of TMC. Therefore, the increase in the effects of TMC on intestinal epithelia did not directly correlate up to the maximum quaternization degree of this polymer, but reached an optimum value already at an intermediate degree of quaternization (ca. 48%). PMID- 12648014 TI - Increasing the aqueous solubility of acetaminophen in the presence of polyvinylpyrrolidone and investigation of the mechanisms involved. AB - It was shown that the aqueous solubility of acetaminophen in the presence of polyvinylpyrrolidone (PVP) increased. The solubility at 25 degrees C increased from 14.3 m mL(-1) in the absence of PVP, to 19.7 mg m(-1) in the presence of 4% w/v PVP, and to 26.7mg mL(-1) in the presence of 8% w/v PVP. Dialysis studies indicated that there is a potential of binding between PVP and acetaminophen in their aqueous solutions. Dialysis studies also revealed that the nature of interaction between PVP and acetaminophen is physical and reversible, and there was no strong binding between PVP and acetaminophen in their solutions. Infrared spectroscopy of acetaminophen/PVP solid dispersion indicated that the mechanism of interaction between PVP and acetaminophen is via hydrogen bonding. Therefore, the increase in solubility of acetaminophen in the presence of PVP is probably attributed to its ability to form a water-soluble complex with PVP. PMID- 12648015 TI - Ketoprofen 1-alkylazacycloalkan-2-one esters as dermal prodrugs: in vivo and in vitro evaluations. AB - Six new 1-alkylazacycloalkan-2-one esters of ketoprofen (1-6) were synthesized and evaluated as potential dermal prodrugs of ketoprofen. Their lipophilicity by both experimental lipophilicity indices (log k') and calculated ClogP was also determined. In vitro experiments were carried out to evaluate the chemical and enzymatic stability and permeation through excised human skin of these new ketoprofen derivatives. Furthermore, we investigated the in vivo topical anti inflammatory activity of ester 5, which showed the best in vitro profile, evaluating the ability of this compound to inhibit methyl nicotinate-induced skin erythema on healthy human volunteers. Esters 1-6 showed increased lipophilicity compared with the parent drug (ketoprofen), good stability in phosphate buffer pH 7.4, and were readily hydrolyzed by porcine esterase. Results from in vitro percutaneous absorption studies showed that, among all esters synthesized, only for esters 1 and 5 did a higher cumulative amount of drug penetrate through the skin, compared with that obtained after topical application of ketoprofen. In vivo results showed an interesting delayed and sustained activity of ester 5, compared with the parent drugs. PMID- 12648016 TI - Enhanced percutaneous permeability of nicardipine hydrochloride by carvone across the rat abdominal skin. AB - The purpose of this study was to investigate the effect of carvone on the permeation of nicardipine hydrochloride across the excised rat abdominal epidermis from 2% w/w hydroxypropyl cellulose (HPC) gel system. The HPC gel formulations containing nicardipine hydrochloride (1% w/w) and selected concentrations of carvone (0 to 12% w/w) were prepared, and evaluated for drug content, stability of the drug, and in vitro permeation of the drug through excised rat abdominal epidermis. The HPC gel was found to contain 99.98 to 101.6% of nicardipine hydrochloride, and the drug was found to be stable in the HPC gels. The permeation flux of nicardipine hydrochloride across rat epidermis was increased markedly by the addition of carvone to the HPC gels. A maximum flux of nicardipine hydrochloride (243.95.70 +/- 1.90 microg/cm2/hr) was observed with an enhancement ratio of 7.9 when carvone was incorporated at a concentration of 12% w/w in the HPC reservoir system. The differential scanning calorimetry and Fourier transform-infrared data indicated that carvone increased the permeability of nicardipine hydrochloride across the rat epidermis by partial extraction of lipids in the stratum corneum. The results suggest that carvone may be useful for enhancing the skin permeability of nicardipine hydrochloride from transdermal therapeutic system containing HPC gel as a reservoir. PMID- 12648017 TI - Synthesis and enzymatic degradation of epichlorohydrin cross-linked pectins. AB - The water solubility of pectin was successfully decreased by cross-linking with increasing amounts of epichlorohydrin in the reaction media. The initial molar ratios of epichlorohydrin/ galacturonic acid monomer in the reaction mixtures were 0, 0.37, 0.56, 0.74, 1.00, 1.47, and 2.44. The resulting epichlorohydrin cross-linked pectins were thus referred to as C-LP0, C-LP37, C-LP56, C-LP75, C LP100, C-LP150, and C-LP250, respectively. Methoxylation degrees ranged from 60.5 +/- 0.9% to 68.0 +/- 0.6%, and the effective cross-linking degrees, determined by quantification of the hydroxyl anions consumed during the reaction, were 0, 17.8, 26.0, 38.3, 46.5, 53.5, and 58.7%. respectively. After incubating the different cross-linked pectins (0.5% w/v) in 25 mL of 0.05 M acetate-phosphate buffer (pH 4.5), containing 50 microL of Pectinex Ultra SP-L (pectinolytic enzymes), between 60 and 80% of the pectin osidic bounds were broken in less than 1 hr. Moreover, increasing the cross-linking degree only resulted in a weak slowing on the enzymatic degradation velocity. PMID- 12648018 TI - Ophthalmic delivery of ciprofloxacin hydrochloride from different polymer formulations: in vitro and in vivo studies. AB - Reservoir-type ocular inserts were fabricated using sodium alginate containing ciprofloxacin hydrochloride as the core (drug reservoir) that was sandwiched between the Eudragit and/or polyvinylacetate films. Ocular inserts were packaged in aluminium foil and sterilized by gamma radiation. These were tested for sterility as per British Pharmacopoeia (BP). Ocular inserts were evaluated for in vitro release rate studies, microbial efficacy, in vivo release studies, efficacy against induced bacterial conjunctivitis in rabbit's eyes, concentration in the aqueous humor, and stability studies as per the International Conference on Harmonization (ICH) guidelines. Ocular inserts passed the test for sterility. They showed zero-order release of the drug in the in vitro and in vivo release studies over a period of 120 hr. The drug was found to be active against selected microorganisms as was proved by microbial efficacy studies. A high correlation coefficient was found between in vitro and in vivo release rate studies. Better improvement was observed in artificially induced bacterial conjunctivitis in rabbit's eyes, compared with marketed eye drops and placebo. Drug concentration in the aqueous humor was found above Minimum Inhibitory Concentration (MIC-90) against selected microorganisms. Shelf-life of the product was found to be more than 2 years. PMID- 12648020 TI - Engineering tools for understanding the hydrodynamics of dissolution tests. AB - In this article, three well-established engineering tools are used to examine hydrodynamics in dissolution testing apparatuses. The application of these tools would provide detailed information about the flow, shear, and homogeneity in dissolution tests. Particle image velocimetry successfully measures two dimensional cross-sections of the velocity field in an experimental device under both laminar and turbulent conditions. The velocity field is also calculated with computational fluid dynamics (CFD), which can rapidly provide data that is difficult or impossible to obtain experimentally. The occurrence of segregated regions within a USP Apparatus II under mild agitation conditions is revealed by CFD simulations and confirmed by laser-induced fluorescence experiments. The results clearly demonstrate that under current operation settings, the USP Apparatus II operates in a regime where the flow is in incipient turbulence, which is a highly time-dependent condition that might explain possible inconsistencies in dissolution results. It is further demonstrated that proposed changes advocating lower speeds or smaller vessels displace the system toward laminar flow conditions characterized by segregation, compromising the robustness of the test and making it vulnerable to variability with respect to sample location. PMID- 12648019 TI - Influence of hydroxypropyl beta-cyclodextrin on the corneal permeation of pilocarpine. AB - The influence of hydroxypropyl beta-cyclodextrin (HPbetaCD) on the corneal permeation of pilocarpine nitrate was investigated by an in vitro permeability study using isolated rabbit cornea. Pupillary-response pattern to pilocarpine nitrate with and without HPbetaCD was examined in rabbit eye. Corneal permeation of pilocarpine nitrate was found to be four times higher after adding HPbetaCD into the formulation. The reduction of pupil diameter (miosis) by pilocarpine nitrate was significantly increased as a result of HPbetaCD addition into the simple aqueous solution of the active substance. The highest miotic response was obtained with the formulation prepared in a vehicle of Carbopol 940. It is suggested that ocular bioavailability of pilocarpine nitrate could be improved by the addition of HPbetaCD. PMID- 12648021 TI - Release of pentoxifylline from xanthan gum matrix tablets. AB - Pentoxifylline-controlled release tablets were prepared using xanthan gum. The effects of polymer concentration, rotation speed, ionic strength, and pH of the dissolution medium on the release of the water-soluble pentoxifylline were studied. The release rate decreased with increasing polymer concentration in the tablet, which was reflected in the increase in the mean dissolution time. A higher rotation speed and increased ionic strength of the dissolution medium resulted in a higher rate of drug release of xanthan-based tablets. A higher release rate of pentoxifylline was also observed using acidic dissolution medium. PMID- 12648022 TI - Skin permeation of 5-methoxypsoralen from topical dosage forms. AB - The topical photochemotherapy of dermatoses with psoralens (PUVA therapy) requires an adequate drug level at the target site (basal epidermis) at the time of UVA radiation. The aim of this work was to enhance 5-methoxypsoralen transport to the basal epidermis, with the goal to shorten the delay between drug application and UVA irradiation. 5-Methoxypsoralen transport through rabbit skin was studied in vitro from topical formulations (water solution, gel. and emulsion). The results obtained show that the use of the emulsion increased the flux through rabbit ear skin, even if partitioning was not favorable. Additionally, the time lag was sensibly reduced, compared with the gel and solution. Furthermore, drug accumulation in human skin in vitro was determined using the thin slicing technique. Human skin accumulation profile was significantly higher for the emulsion, compared with the gel, indicating that the delay between psoralen application and UVA irradiation can be shortened. PMID- 12648023 TI - Maternal-fetal transport of hypoglycaemic drugs. AB - Due to legal, ethical and monetary problems, drug studies in pregnancy are rare. Numerous pharmacokinetic and pharmacodynamic changes occur in pregnancy that can affect the efficacy and safety of drugs, and these are difficult to predict without appropriate studies. Drugs potentially useful and safe in pregnancy have to either not cross the placenta and/or be harmless to the fetus at clinically relevant concentrations. The first characteristic can be predicted using in vitro models such as the placenta perfusion model. In the case of glibenclamide (glyburide), in vitro experiments showed minimal maternal-fetal transfer, leading to completion of a successful clinical trial of this drug in gestational diabetes. Insulin, the main drug used in diabetes during pregnancy, has also been shown not to cross the placenta in vitro, as has insulin lispro. Animal insulin may cross the placenta when complexed with anti-insulin antibodies. Other sulphonylurea drugs (tolbutamide and chlorpropamide) have been shown to cross the placenta both in vitro and in vivo and to produce toxicity in the fetus. This review summarises the pharmacokinetic data available for hypoglycaemic drugs during pregnancy, as well as the potential role for the in vitro placenta perfusion model in the preclinical evaluation of drugs with potential usefulness in pregnancy. PMID- 12648024 TI - Pharmacokinetic principles of bacteriophage therapy. AB - Use of bacteriophage to control bacterial infections, including antibiotic resistant infections, shows increasing therapeutic promise. Effective bacteriophage therapy requires awareness of various novel kinetic phenomena not known in conventional drug treatments. Kinetic theory predicts that timing of treatment could be critical, with the strange possibility that inoculations given too early could be less effective or fail completely. Another paradoxical result is that adjuvant use of an antibiotic can sometimes diminish the efficacy of phage therapy. For a simple kinetic model, mathematical formulae predict the values of critical density thresholds and critical time points, given as functions of independently measurable biological parameters. Understanding such formulae is important for interpreting data and guiding experimental design. Tailoring pharmacokinetic models for specific systems needs to become standard practice in future studies. PMID- 12648026 TI - Pharmacokinetics of selective estrogen receptor modulators. AB - Selective estrogen receptor modulators (SERMs) are a class of compounds used to treat and prevent breast cancer and osteoporosis. SERMs currently approved for use in patients include tamoxifen, toremifene and raloxifene. These compounds are well tolerated in patients, and the most common adverse effects experienced in patients undergoing SERM therapy include vasomotor symptoms such as hot flashes and vaginal discharge. New SERMs currently under development for use in the treatment and prevention of osteoporosis and breast cancer include ospemifene, a derivative of toremifene, and arzoxifene, a compound very similar in structure to raloxifene. SERMs are administered orally at doses ranging from 20 to 60 mg/day. Tamoxifen and toremifene have a bioavailability of approximately 100%, whereas that of raloxifene is only 2%. SERMs are very highly bound to plasma proteins (>95%). Tamoxifen and toremifene are metabolised by the cytochrome p450 enzyme system, and raloxifene is metabolised by glucuronide conjugation. The terminal elimination half-lives of these drugs range from 27.7 hours to 7 days. The pharmacokinetics of these compounds are affected in hepatically impaired patients, but not in renally impaired patients. SERMs have several potential drug interactions with other agents, such as warfarin, rifampicin (rifampin), cholestyramine and aromatase inhibitors. PMID- 12648027 TI - Pharmacokinetics of intravenous and rectal ketoprofen in young children. AB - OBJECTIVE: To evaluate the relative bioavailabilities of ketoprofen after intravenous and rectal administration to young children. DESIGN: Open-label prospective parallel-group study. PATIENTS: Participants were 28 children aged 7 to 93 months. METHODS: Eighteen children received a single intravenous injection of ketoprofen 1 mg/kg, and ten children, weight 16-24 kg, received a 25mg ketoprofen suppository. Venous blood samples were collected at selected times after administration, ranging from 2 minutes to 8 hours for the intravenous group and from 30 minutes to 8 hours for the suppository group. A validated high performance liquid chromatography method was used to measure plasma ketoprofen concentrations. RESULTS: In the intravenous group, the maximum plasma concentration of ketoprofen ranged between 10.5 and 22.2 mg/L, and in the suppository group, following dose normalisation to 1 mg/kg of ketoprofen, between 3.8 and 7.4 mg/L. In the intravenous group, area under the concentration-time curve from zero to infinity ranged between 9.2 and 23.5 mg x h/L, and in the suppository group after dose normalisation between 8.8 and 12.9 mg x h/L. The bioavailability of ketoprofen from the suppository was about 73%. Volume of distribution was 0.04-0.10 L/kg in the intravenous group and 0.08-0.16 L/kg in the suppository group. The terminal half-life was comparable in both study groups, ranging between 0.7 and 3.0 hours in the intravenous group and between 1.2 and 2.9 hours in the suppository group. CONCLUSION: Absorption of ketoprofen after rectal administration is reasonably rapid and predictable. Because the bioavailability of rectal ketoprofen is also relatively high, a suppository may be used in children in whom the drug cannot be given intravenously or by mouth. PMID- 12648028 TI - Influence of age on the pharmacokinetics and pharmacodynamics of ximelagatran, an oral direct thrombin inhibitor. AB - OBJECTIVE: To investigate the influence of age on the pharmacokinetics and pharmacodynamics of ximelagatran. STUDY DESIGN: This was an open-label, randomised, 3 x 3 crossover study with 4 study days, separated by washout periods of 7 days. SUBJECTS: Subjects comprised 6 healthy young men (aged 20-27 years) and 12 healthy older men and women (aged 56-70 years). METHODS: All subjects received a 2mg intravenous infusion of melagatran over 10 minutes followed, in randomised sequence, by a 20 mg immediate-release tablet of ximelagatran with breakfast, a 20 mg immediate-release tablet of ximelagatran while fasting, and a 7.5 mg subcutaneous injection of ximelagatran. The primary variables were the plasma concentration of melagatran, the active form of ximelagatran, and the activated partial thromboplastin time (APTT), an ex vivo coagulation time measurement used to demonstrate inhibition of thrombin. RESULTS: After oral and subcutaneous administration, ximelagatran was rapidly absorbed and biotransformed to melagatran, its active form and the dominant compound in plasma. The metabolite pattern in plasma and urine was similar in young and older subjects after both oral and subcutaneous administration of ximelagatran clearance of melagatran was correlated with renal function, resulting in about 40% (after intravenous melagatran) to 60% (after oral and subcutaneous ximelagatran) higher melagatran exposure in the older than in the young subjects. Renal clearance of melagatran, was 7.7 L/h and 4.9 L/h in the younger and older subjects, respectively. The interindividual variability inn the area under the melagatran plasma concentration-time curve was low following all regimens (coefficient variation 12-25%). The mean bioavailability of melagatran in young and older subjects was approximately 18 and 12% , respectively, following oral administration of ximalagratan, and 38 and 45%, respectively, following subcutaneous administration of ximelagatran. The bioavailability of melagatran following oral administration of ximelagatran was unaffected by whether subjects were fed or fasting, although the plasma concentration of melagatran peaked about 1 hour later under fed than fasting conditions, due to gastric emptying of the immediate-release tablet formulation used. The APTT as prolonged with increasing melagatran plasma concentration-effect relationship was independent of age. CONCLUSIONS: There were no age-dependent differences in the absorption and biotransformation of ximelagatran, and the observed differences in exposure to melagatran can be explained by differences in renal function between the young and older subjects. PMID- 12648029 TI - Pharmacokinetics of methylphenidate after oral administration of two modified release formulations in healthy adults. AB - OBJECTIVE: To compare the rate and extent of absorption of DL-threo methylphenidate (MPH) from two modified-release MPH formulations at their respective recommended starting doses in healthy adult volunteers. DESIGN: Open label, randomised, crossover, bioavailability study. PARTICIPANTS: Twenty healthy adult male and female volunteers. METHODS: Subjects received single doses of two modified-release formulations of MPH, a 20mg capsule (Ritalin) LA) and an 18 mg tablet (Concerta). A total of 19 plasma samples was collected over 24 hours, and MPH plasma concentrations were determined by liquid chromatography-mass spectrometry (LC-MS/MS). These values were used to calculate standard noncompartmental pharmacokinetic parameters describing the rate (peak concentration and time to peak concentration) and extent (area under the concentration-time curve, AUC) of absorption of the two formulations. The relative bioavailability of the two drugs was assessed using a 90% confidence interval, based on the lower and upper endpoints of the confidence interval for the ratios of the geometric means (log transformed) being within the 0.80-1.25 equivalence criterion. RESULTS: Nineteen subjects, ten male and nine female, aged 21-34 years completed both treatment phases of the study. The Ritalin LA formulation displayed a distinctly biphasic pharmacokinetic profile, with mean initial peak plasma concentration of 7 microg/L at an average of 2.1 hours after administration and a second peak of 9.3 microg/L occurring at 5.6 hours. In contrast, the profile of the Concerta formulation rapidly reached an initial plateau concentration of 3.4 microg/L at 3.3 hours after administration and a second mean plateau concentration of 5.9 microg/L approximately 6 hours after administration. Substantially more MPH was absorbed from Ritalin LA than from Concert over the first 4 hours; the respective AUC(4) values were 18.5 and 9.3 microg x h/L (p < 0.001). The overall extent of absorption of MPH was similar between the two formulations. Oral clearance was identical between the two dosage forms. CONCLUSIONS: The Ritalin LA formulation exhibited more rapid initial absorption and reached significantly higher peak plasma concentrations compared with the Concerta formulation, although the oral bioavailability of MPH was similar between the two formulations. The Ritalin LA capsule demonstrated a distinctly bimodal plasma concentration-time profile. MPH plasma concentrations resulting from Concerta reached a peak at 6 hours. These results indicate that the recommended starting dose of the Ritalin LA 20 mg capsule formulation provides more rapid absorption and higher peak plasma concentrations than the recommended 18 mg starting dose of the Concerta formulation. PMID- 12648030 TI - A model to assess the cost effectiveness of statins in achieving the UK National Service Framework target cholesterol levels. AB - BACKGROUND: Coronary heart disease (CHD) is a public health priority in the UK. The National Service Framework (NSF) has set standards for the prevention, diagnosis and treatment of CHD, which include the use of cholesterol-lowering agents aimed at achieving targets of blood total cholesterol (TC) < 5.0 mmol/L and low density lipoprotein-cholesterol (LDL-C) < 3.0 mmol/L. In order to achieve these targets cost effectively, prescribers need to make an informed choice from the range of statins available. AIM: To estimate the average and relative cost effectiveness of atorvastatin, fluvastatin, pravastatin and simvastatin in achieving the NSF LDL-C and TC targets. DESIGN: Model-based economic evaluation. METHODS: An economic model was constructed to estimate the number of patients achieving the NSF targets for LDL-C and TC at each dose of statin, and to calculate the average drug cost and incremental drug cost per patient achieving the target levels. The population baseline LDL-C and TC, and drug efficacy and drug costs were taken from previously published data. Estimates of the distribution of patients receiving each dose of statin were derived from the UK national DIN-LINK database. RESULTS: The estimated annual drug cost per 1000 patients treated with atorvastatin was pound 289000, with simvastatin pound 315000, with pravastatin pound 333000 and with fluvastatin pound 167000. The percentages of patients achieving target are 74.4%, 46.4%, 28.4% and 13.2% for atorvastatin, simvastatin, pravastatin and fluvastatin, respectively. Incremental drug cost per extra patient treated to LDL-C and TC targets compared with fluvastatin were pound 198 and pound 226 for atorvastatin, pound 443 and pound 567 for simvastatin and pound 1089 and pound 2298 for pravastatin, using 2002 drug costs. CONCLUSIONS: As a result of its superior efficacy, atorvastatin generates a favourable cost-effectiveness profile as measured by drug cost per patient treated to LDL-C and TC targets. For a given drug budget, more patients would achieve NSF LDL-C and TC targets with atorvastatin than with any of the other statins examined. PMID- 12648031 TI - An economic analysis of the Atorvastatin Comparative Cholesterol Efficacy and Safety Study (ACCESS). AB - INTRODUCTION: The objective of the Atorvastatin Comparative Cholesterol Efficacy and Safety Study (ACCESS) was to compare the efficacy and safety of the five 3 hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors in a randomised, controlled, yet large-scale study. ACCESS also produced data that permitted comparative analysis of the cost to achieve National Cholesterol Education Panel (NCEP) II low density lipoprotein-cholesterol (LDL-C) targets. STUDY DESIGN: A 54-week, multicentre, open-label, randomised, parallel-arm, active-control study in men and women with or without documented coronary heart disease or peripheral vascular disease. Data included medication use, clinic visits, adverse events, LDL-C and other laboratory measures. Analyses of resource use and cost are reported from a third-party payer perspective. METHODS: Patients were randomly assigned to receive one of the following treatments: atorvastatin (10-80 mg/day); fluvastatin (20-40 mg/day, or 40 mg twice daily); lovastatin (20 40 mg/day, or 40 mg twice daily); pravastatin (10-40 mg/day); or simvastatin (10 40 mg/day). Patients were started at the lowest available dose and titrated to higher doses at 6-week intervals until they achieved the NCEP II LDL-C target or reached the highest available dose of medication. PATIENTS: A total of 153 centres enrolled 3887 patients: atorvastatin (n = 1944); fluvastatin (n = 493); lovastatin (n = 494); pravastatin (n = 478); and simvastatin (n = 478). Inclusion criteria included LDL-C >or= 30 mg/dL higher than NCEP II LDL-C target (stratified by risk factors), fasting triglyceride values < 400 mg/dL, and a confirmed negative serum pregnancy test. Known hypersensitivity to statins, use of prohibited medications, uncontrolled diabetes, acute liver disease and age > 80 years or < 18 years were among the exclusion criteria. RESULTS: Mean total treatment costs to reach LDL-C targets for patients receiving atorvastatin (US dollars 683.37 in 2001) were significantly less than mean total treatment costs for patients receiving fluvastatin (difference = US dollars 211.35, p < 0.01), lovastatin (US dollars 607.96, p < 0.01), pravastatin (US dollars 424.60, p < 0.01) and simvastatin (US dollars 95.74, p < 0.01). Results were robust to sensitivity analyses using alternative definitions of the patient population (randomised, intent-to-treat, completers) and cost measures (50th percentile charges, 95th percentile charges, Medicare prices). CONCLUSIONS: Compared with the other statins studied, atorvastatin was associated with the lowest resource use and costs when used to treat patients to their NCEP II LDL-C targets. Atorvastatin was also associated with the highest percentage of patients achieving their desired clinical outcomes. Therefore, in cost-effectiveness terms, it dominated the four other statins. PMID- 12648032 TI - A pharmacoeconomic evaluation of the Myocardial Ischaemia Reduction with Aggressive Cholesterol Lowering (MIRACL) study in the United Kingdom. AB - OBJECTIVE: To determine the short-term healthcare costs associated with intensive lipid lowering with atorvastatin initiated within 24-96 hours of the occurrence of acute coronary syndrome (ACS) in patients in the UK. METHODS: Patient-level clinical outcome data from the Myocardial Ischaemia Reduction with Aggressive Cholesterol Lowering (MIRACL) trial and standard cost data were used to compare the total expected 16-week cost per patient on atorvastatin 80 mg/day versus placebo. Clinical outcomes assessed included the following: death; cardiac arrest with resuscitation; nonfatal myocardial infarction; worsening angina pectoris with objective evidence of myocardial ischaemia requiring rehospitalisation; surgical or percutaneous coronary revascularisation; nonfatal stroke; hospitalisation for angina without objective evidence of myocardial ischaemia; and new or worsening congestive heart failure requiring rehospitalisation. All relevant direct medical costs from the perspective of the NHS were considered. RESULTS: The total expected cost was pound 784.05 per patient in the placebo cohort and pound 851.59 per patient in the atorvastatin cohort, resulting in an incremental cost of pound 67.54 per patient in the atorvastatin group. The cost per event avoided was pound 1762.04. A third of the cost of atorvastatin treatment was offset within 16 weeks by the cost savings resulting from the reduction in the number of events in the atorvastatin cohort compared with the placebo cohort. CONCLUSION: The clinical benefits of short-term intensive atorvastatin treatment administered after ACS is attainable through a marginal increase in 'upfront' costs. PMID- 12648033 TI - Financial and health costs of uncontrolled blood pressure in the United Kingdom. AB - BACKGROUND: Uncontrolled hypertension is associated with an elevated risk of all cardiovascular disease, especially stroke. However, many patients with hypertension do not achieve agreed targets for blood pressure. OBJECTIVE: To estimate the cost and morbidity consequences of uncontrolled hypertension in the UK. DESIGN: Descriptive epidemiological study. METHODS: The study used a burden of-disease model combining data on the prevalence of hypertension, the incidence of major cardiovascular (CV) events and the costs of treating these events. The prevalence of uncontrolled hypertension was taken from the 1998 Health Survey for England. The incidences of three CV events, acute myocardial infarction, congestive heart failure and stroke, at different levels of achieved blood pressure, were estimated using results from the Hypertension Optimal Treatment study. Costs were taken from published sources. We estimated the number of major CV events and acute hospital costs that would be avoided if all people with hypertension had blood pressure treated to target levels. RESULTS: The model estimated that in the UK 5.7 million adults (12% of the population aged >16 years) have actual blood pressure above 160/95 mm Hg, and a further 10.3 million (21%) have blood pressure in the range 140/90-160/95 mm Hg. An estimated 58000 major CV events per year occur in these patients that would be avoided if their blood pressure was at target levels. If all patients had blood pressure treated to target, the cost to the NHS of managing major CV events would fall by pound 97.2 million per year at 2000/01 prices (95% CI: pound 56- pound 144 million). CONCLUSION: Failure to achieve blood pressure targets contributes substantially to avoidable NHS costs and to the number of CV events in the UK. PMID- 12648025 TI - Pharmacokinetics and pharmacodynamics of cannabinoids. AB - Delta(9)-Tetrahydrocannabinol (THC) is the main source of the pharmacological effects caused by the consumption of cannabis, both the marijuana-like action and the medicinal benefits of the plant. However, its acid metabolite THC-COOH, the non-psychotropic cannabidiol (CBD), several cannabinoid analogues and newly discovered modulators of the endogenous cannabinoid system are also promising candidates for clinical research and therapeutic uses. Cannabinoids exert many effects through activation of G-protein-coupled cannabinoid receptors in the brain and peripheral tissues. Additionally, there is evidence for non-receptor dependent mechanisms. Natural cannabis products and single cannabinoids are usually inhaled or taken orally; the rectal route, sublingual administration, transdermal delivery, eye drops and aerosols have only been used in a few studies and are of little relevance in practice today. The pharmacokinetics of THC vary as a function of its route of administration. Pulmonary assimilation of inhaled THC causes a maximum plasma concentration within minutes, psychotropic effects start within seconds to a few minutes, reach a maximum after 15-30 minutes, and taper off within 2-3 hours. Following oral ingestion, psychotropic effects set in with a delay of 30-90 minutes, reach their maximum after 2-3 hours and last for about 4-12 hours, depending on dose and specific effect. At doses exceeding the psychotropic threshold, ingestion of cannabis usually causes enhanced well-being and relaxation with an intensification of ordinary sensory experiences. The most important acute adverse effects caused by overdosing are anxiety and panic attacks, and with regard to somatic effects increased heart rate and changes in blood pressure. Regular use of cannabis may lead to dependency and to a mild withdrawal syndrome. The existence and the intensity of possible long-term adverse effects on psyche and cognition, immune system, fertility and pregnancy remain controversial. They are reported to be low in humans and do not preclude legitimate therapeutic use of cannabis-based drugs. Properties of cannabis that might be of therapeutic use include analgesia, muscle relaxation, immunosuppression, sedation, improvement of mood, stimulation of appetite, antiemesis, lowering of intraocular pressure, bronchodilation, neuroprotection and induction of apoptosis in cancer cells. PMID- 12648036 TI - Abstracts of the 37th annual scientific meeting of the European Society for Clinical Investigation. Verona, Italy, 2-5 April 2003. PMID- 12648034 TI - The economic burden of stroke in the United Kingdom. AB - AIM: To estimate the cost of treating stroke in the UK. METHODS: A cost-of illness model was constructed to estimate stroke-related costs over a 5-year period. The cost estimates were based on data from a large, randomised, prospective study comparing alternative strategies of stroke care. The study collected detailed data on resource use in hospital, primary care, healthcare contacts, and utilisation of social services over a period of 1 year following stroke. A Markov framework was used to extrapolate 1-year costs over 5 years. RESULTS: The model estimated that, for every patient who experiences a stroke, the cost to the NHS in the UK is pound 15306 over 5 years and, when informal care costs are included, the amount increases to pound 29405 (2001/2002 prices). The robustness of the cost findings was explored with the use of sensitivity analysis. This focused on the key variables of rates of recurrent stroke, the estimated acute costs, and costs attached to institution and home care. CONCLUSION: As well as being a considerable cause of morbidity and mortality, stroke is also a huge cost burden to both the UK's NHS and the carers of stroke victims. PMID- 12648039 TI - Let's paddle this boat. PMID- 12648035 TI - Organization on the plasma membrane of the retinitis pigmentosa protein RP2: investigation of association with detergent-resistant membranes and polarized sorting. AB - Mutations in the retinitis pigmentosa protein gene RP2 account for up to 15% of X linked retinitis pigmentosa. RP2 is a novel protein of unknown function, which is targeted to the plasma membrane by dual N-terminal acyl-modification. Dual acylated proteins are targeted to lipid rafts, and some are subject to polarized sorting. Therefore we investigated the organization of RP2 on the plasma membrane. Endogenous RP2 protein was predominantly localized at the plasma membrane, and exogenously expressed green-fluorescent-protein-tagged protein was also targeted to the membrane in a wide range of cultured cells. High levels of endogenous RP2 protein were present in HeLa cells and in the retinal pigment epithelium-derived cell line ARPE19. A significant proportion of RP2 in cultured neuroblastoma cells was associated with detergent-resistant membranes (DRMs), but much less than other dually acylated proteins (e.g. Lyn and Fyn). In contrast, the RP2-interacting protein Arl3 (ADP-ribosylation factor-like 3) was not found to be associated with DRMs. The association of RP2 with DRMs was cholesterol dependent. In polarized epithelial cells in culture and in vivo, RP2 was present in both the apical and basolateral domains of the plasma membrane. These data show that RP2 is not specific to either domain, unlike some other dually acylated proteins. Interestingly, the level of RP2 protein increased in the epithelial cell line Caco-2 with differentiation and polarization. These data show that RP2 is present on the membrane of all cell types examined both in vitro and in vivo, and that RP2 associates with lipid rafts, suggesting a potential role for the protein in signal transduction. PMID- 12648038 TI - Keratocytes: more than a framework for the window. PMID- 12648040 TI - Retinal detachment in cytomegalovirus retinitis: intravenous versus intravitreal therapy. AB - PURPOSE: To compare the rate of retinal detachment in a group of patients treated with intravitreal ganciclovir to the rate of retinal detachment in a group of patients treated with systemic ganciclovir. METHODS: A retrospective non randomized comparative case series was conducted. The participants were 186 consecutive patients with cytomegalovirus (CMV) retinitis treated at two centres over the period from June 1990 to May 1997. Of the 186, 133 patients had systemic therapy of whom 113 had intravenous ganciclovir or foscarnet and 20 had oral ganciclovir, and 53 patients had intravitreal induction and maintenance therapy. The main outcome measure was the rate of retinal detachment, which was calculated using person-months-at-risk denominators. The effect measure used in the main comparative analysis was the hazard ratio obtained from multiple regression using the Cox proportional hazards model. RESULTS: Retinal detachment occurred in three of 53 patients in the intravitreal therapy group, one of 20 patients receiving oral maintenance therapy and 21 of 113 patients receiving intravenous maintenance therapy. The risk of retinal detachment with systemic therapy was 14-fold higher than with intravitreal therapy (P < 0.001)and up to fourfold higher with oral maintenance therapy. CONCLUSIONS: Intravitreal therapy offers a significant advantage over systemic therapy in the treatment of CMV retinitis by substantially reducing the risk of CMV-related retinal detachment. PMID- 12648041 TI - Limited macular translocation for subfoveal choroidal neovascularization in age related macular degeneration. AB - PURPOSE: To examine the safety and efficacy of limited macular translocation followed by laser photocoagulation in the management of subfoveal choroidal neovascularization (CNV) in age-related macular degeneration (AMD). METHODS: A prospective study was conducted on eight consecutive patients undergoing limited macular translocation followed by laser photocoagulation for the treatment of subfoveal classic CNV form of AMD. Patients were followed up for a minimum of 12 months. The magnitude of foveal translocation, visual outcomes and complications were assessed. RESULTS: In all cases the fovea was successfully translocated inferiorly, with a median displacement of 1.1 mm. Thermal laser photocoagulation was subsequently performed in all cases. Visual acuity improved in five eyes, remained unchanged in two eyes and worsened in one eye. The mean improvement in visual acuity was by 0.19 logMAR. At 12 months, six of the eight patients (75%) achieved Snellen visual acuity of at least 6/15, with four patients (50%) achieving Snellen visual acuity of 6/9. CONCLUSION: In this small case series, limited macular translocation was found to be an effective and reproducible means of treating small well-defined subfoveal CNV. PMID- 12648042 TI - Golf-related ocular injuries. AB - PURPOSE: To examine patient outcomes from golf-related ocular injuries. METHODS: A retrospective case review was undertaken of subjects with golf-related ocular injuries treated at the Auckland and Waikato Hospitals during the 5-year period from 1997 to 2002. RESULTS: Of 11 injuries treated in the 5-year period, five of the subjects incurred globe rupture with two subjects requiring enucleation. Seven of the injuries were from golf balls and four injuries were from golf clubs. Six of the subjects had a final visual acuity of 6/48 or worse. CONCLUSION: Golfing injuries, though uncommon, are frequently associated with severe ocular trauma and poor visual outcome. Although unpredictable and difficult to avoid, proper golf etiquette and safety may diminish the incidence of golf-related ocular injures. Injuries caused by golf clubs occurred only in children. PMID- 12648043 TI - The New Zealand cataract and refractive surgery survey 2001. AB - BACKGROUND: This study documents the current practice for cataract and refractive surgery in New Zealand. METHODS: A postal questionnaire was distributed late in 2000 to all consultant ophthalmologists in New Zealand. Most questions were identical to the 2000 survey of the American Society of Cataract and Refraction Surgeons (ASCRS) enabling a comparison. RESULTS: There was an 84% return rate from the 103 surveys distributed. In regards to cataract surgery, topical anaesthesia was used by 12% of respondents, clear corneal incisions by 64%, no suture was used by 94%, diamond blades were used by 33%, disposable blades were reused by 64%, preoperative antibiotics were used by 28%, anti-biotic was used in the irrigant by 10%, postoperative injections of steroid/antibiotics were used by 63% and 41% of patients had three postoperative visits after cataract surgery. In regards to refractive surgery, 51% of respondents had access to an excimer laser and clear lens extraction was performed by 13 ophthalmologists. Advice to a 30 year-old -7.00 myope wanting refractive surgery was to have LASIK (88%), wait (8%) or have no surgery (4%). DISCUSSION: In broad terms, New Zealand ophthalmologists' cataract and refractive practice is similar to that of the members of ASCRS with the exception of the use of topical anaesthetic (NZ 12%vs USA 49%) and the higher use of postoperative injections of steroid and antibiotic at the end of surgery (NZ 63%vs USA 20%). PMID- 12648044 TI - Comparison of the Zeiss IOLMaster and applanation A-scan ultrasound: biometry for intraocular lens calculation. AB - PURPOSE: A comparison of axial length estimates using applanation A-scan ultrasound and the Zeiss IOLMaster was conducted. The accuracy in predicting postoperative refraction determined by each method was also compared. METHODS: A cross-sectional study was performed on 51 eyes in 46 patients presenting to clinical practice for cataract surgery assessment. Preoperative measurement of axial length was performed with applanation ultrasound and the IOLMaster. The IOLMaster measurements were used to determine the intraocular lens power based on the SRK/T formula. Postoperative refractive outcomes were obtained and spherical equivalent calculated and compared to the predicted refractive error with each biometric method. RESULTS: On average the axial lengths measured by the IOLMaster were longer by 0.15 mm compared to ultrasound biometry (P < 0.01). Using the IOLMaster over applanation ultrasound biometry significantly improved the postoperative refractive outcome from 0.65 D to 0.42 D (P = 0.011). CONCLUSIONS: The IOLMaster provides an accurate axial length measurement and results in accurate intraocular lens power calculation based on the SRK/T formula. It is quick and easy to use and provides a non-contact technique with no risk of infection or corneal abrasion. PMID- 12648046 TI - Otago Glaucoma Surgery Outcome Study. Atypical late failure of drainage following clear cornea trabeculectomy. AB - BACKGROUND: Bleb scarring is the major cause of late drainage failure following the standard trabeculectomy. To overcome this, Cairns described a modification of his original technique, which he termed the clear cornea trabeculectomy. The essential feature was communication between the anterior chamber and subconjunctival space without intraoperative disturbance of the conjunctival or Tenon's tissue. As there is little published data on Cairns' clear cornea trabeculectomy, this study aimed to assess the long-term success of this procedure relative to conventional trabeculectomy. METHODS: Patients who underwent clear cornea trabeculectomy at Dunedin Hospital were identified. Cases in which late failure occurred were subject to a detailed review. Records of all patients in whom late failure of drainage occurred following conventional trabeculectomy performed since 1985 were also reviewed. The nature of late drainage failure associated with these different procedures was assessed. RESULTS: Late failure of drainage occurred in three of 21 eyes following clear cornea trabeculectomy. In each case there was an accelerated drainage failure with a dramatic rise in intraocular pressure. Consequent visual loss occurred in two patients. Late failure occurred in 32 of the 450 cases of conventional trabeculectomy, in all of which the pressure rise was gradual and less acute. CONCLUSIONS: This report highlights a previously unreported type of late failure associated with clear cornea trabeculectomy, which may result in significant visual loss. This reinforces the view that the standard Cairns' trabeculectomy is a safer procedure. PMID- 12648045 TI - Topical anaesthesia: a risk factor for post-cataract-extraction endophthalmitis? AB - PURPOSE: To investigate if the incidence of postoperative endophthalmitis in temporal clear corneal cataract surgery is influenced by the use of topical anaesthesia compared to retrobulbar anaesthesia. METHODS: A retrospective study was conducted of one surgeon's cataract surgery between October 1997 and October 2001. RESULTS: Between October 1997 and May 2000, there were 633 cataract extractions performed with 219 patients operated under topical anaesthesia and 414 under retro-bulbar injection. The selection criteria were that surgically easier cases underwent topical anaesthesia. There were five patients who suffered postoperative endophthalmitis in their first week, of which four cases were under topical anaesthesia. This was shown to be borderline significance of P = 0.05 using the Fischer exact 2-tailed test. There was a complicated case, operated under retrobulbar anaesthesia, who had a low-grade endophthalmitis in the second postoperative week. The four topical cases and the case from the second week all grew Staphylococcus epidermidis. The retrobulbar case developing endophthalmitis in the first week grew alpha haemolytic Streptococcus. After May 2000, there was a change to performing all cataract surgery under retrobulbar anaesthesia and the next 453 cases had no incidence of endophthalmitis. CONCLUSION: Topical anaesthesia techniques in temporal clear corneal cataract extraction may be a factor in endophthalmitis. PMID- 12648047 TI - Fornix-based trabeculectomy using the 'anchoring' corneal suture technique. AB - AIM: To assess the safety and efficacy of fornix-based tra-beculectomy with the 'anchoring' corneal suture technique in Chinese patients. METHODS: A retrospective non-comparative case series of 66 eyes of 63 Chinese patients who underwent fornix-based trabeculectomy combined with intraoperative mitomycin C (MMC) 0.4 mg/mL for 2-5 min were studied. Conjunctival wound closure of all eyes was with the anchoring corneal suture technique using a 9-0 nylon suture to prevent leakage at the corneolimbal interface. The success rate, with or without anti-glaucoma medication, the complication rate and the longevity of the drainage blebs were analyzed. RESULTS: At a mean follow-up period of 447.36 +/- 337.98 days, the mean intraocular pressure decreased from 26.83 +/- 7.90 mmHg to 17.74 +/- 8.74 mmHg (P < 0.001). The mean glaucoma medication decreased from 2.95 +/- 0.98 preoperatively to 1.23 +/- 1.41 postoperatively (P < 0.001). Thirty-one eyes (47.0%) required no anti-glaucoma medication postoperatively. Twenty eyes (30.3%) had qualified success postoperatively. Eight eyes (12.1%) had early wound leakage that resolved with conservative treatment. Three eyes (4.5%) required surgical repair. One eye (1.5%) had hypotonous maculopathy. No other major complications were encountered. CONCLUSION: Fornix-based trabeculectomy with adjunctive mitomycin C employing the anchoring corneal suture technique appears to be both safe and effective in Chinese patients. PMID- 12648048 TI - Blepharophimosis: a recommendation for early surgery in patients with severe ptosis. AB - PURPOSE: To determine the optimal age for surgical correction of blepharophimosis. Associated features and their effects on incidence of amblyopia were also investigated. METHODS: The study was a retrospective case series of 28 patients with blepharophimosis, ptosis and epicanthus inversus syndrome presenting to a tertiary referral eyelid, lacrimal and orbital clinic. RESULTS: Amblyopia was present in 39% of patients. Patients with coexistent strabismus had a 64% incidence of amblyopia compared to 24% for those without strabismus. Hypermetropia was present in 43% of patients and 7% were myopic. Significant astigmatism was found in 40% of patients, but these factors did not increase the risk of amblyopia. Patients with severe ptosis had lower rates of amblyopia than those with moderate ptosis but had their ptosis corrected at a median age of 2 years compared to 5 years for those with moderate ptosis. There was an 18% incidence of nasolacrimal drainage problems. A good to excellent cosmetic outcome was achieved in 86% of patients. A positive family history was noted in 75% of patients, usually with paternal inheritance. CONCLUSIONS: Patients with blepharophimosis have a high rate of amblyopia. Co-existent strabismus doubles the risk of amblyopia. Ptosis alone causes mild to moderate amblyopia only. Patients with severe ptosis should have their ptosis corrected before 3 years of age, and all other patients should undergo surgery before 5 years of age. PMID- 12648049 TI - Toxocara canis: egg presence in Melbourne parks and disease incidence in Victoria. AB - PURPOSE: Toxocara canis can cause blinding eye disease. This study assessed the presence of T. canis eggs in soil from parks in Melbourne and also the incidence of presumed ocular toxocariasis in Victoria. METHODS: One hundred and eighty soil samples were collected from nine suburban locations in Melbourne, Australia. These were analyzed for the presence of T. canis eggs. A search of laboratory records of T. canis serology requests from Victorian patients over an 8-year period was performed. RESULTS: Only one soil sample was positive for T. canis eggs. Positive T. canis serology was reported in 13 samples from patients. These patients all had ocular features suggestive of T. canis infection. CONCLUSION: Toxocara canis eggs are rare in public parks in Melbourne and symptomatic ocular toxocariasis is uncommon in the Victorian population. The acquisition of the disease is unlikely to be from public parks. PMID- 12648050 TI - Confocal imaging of the human keratocyte network using the vital dye 5 chloromethylfluorescein diacetate. AB - BACKGROUND: The human corneal stroma consists of intercalated layers of collagen and keratocytes. These cells are known to maintain the stroma and aid in repair but it is likely they have other crucial roles throughout the cornea. The complexity of their anatomy is revealed in this study by ex vivo in situ images of the human keratocyte covering a range of ages. METHODS: Human donor corneas of different ages were stained with 5-chloromethylfluorescein diacetate (CMFDA), a dye that is anchored and retained within the cell cytoplasm. The tissue was fixed, sectioned, mounted, and then imaged using a confocal laser scanning microscope at various magnifications and tissue planes. The digital image sets were transferred to multifunction image processing software for analysis and production of 3-D stereo images of keratocyte networks throughout the stroma. RESULTS: High quality images of CMFDA-stained cells revealed differences in the structure and orientation of keratocytes in the anterior, central and posterior stroma, which did not differ throughout the age-range studied. This method reveals very fine cell process ramifications not previously visualized, orientated in lateral and antero-posterior directions, and it confirms the potential for multidirectional communication between keratocyte networks. CONCLUSIONS: This qualitative study found consistency of keratocyte morphology in the normal human cornea throughout life. It confirmed differences in keratocyte anatomy, and the potential for rapid cellular communication by multiple interconnecting processes supporting cohesive keratocyte activity. This high resolution 3-D microscopic study should assist in identifying gross deviant cellular behaviour in post-surgical and disease states. PMID- 12648051 TI - Pseudo-hypopyon as the presenting feature in B-cell and T-cell intraocular lymphoma. AB - The clinicopathologic correlation of two patients with primary intraocular lymphoma is described, both of whom had a rare and unusual presentation of hypopyon uveitis. Sampling of aqueous fluid proved the cytopathologic diagnosis of B-cell lymphoma in the first patient. In the second patient an iris biopsy confirmed a T-cell lymphoma. The samples were examined using haematoxylin and eosin, and immunohistochemical stains. These patients were diagnosed with primary intraocular lymphoma on initial presentation with hypopyon uveitis. Secondary glaucoma was noted in the patient with T-cell lymphoma. Both cases appeared confined to the eye and initially responded favourably to aggressive chemotherapy and radiotherapy, but later went on to develop systemic involvement. Clinicians should be aware of this dramatic mode of presentation, which is unusual for ocular lymphoma. PMID- 12648052 TI - Fungal granuloma following ruthenium plaque radiotherapy of a choroidal melanoma. PMID- 12648053 TI - Acute vitreomacular traction with early spontaneous resolution. PMID- 12648054 TI - Idiopathic polypoidal choroidal vasculopathy as a cause of a disciform macular scar. PMID- 12648055 TI - Cystoid macular oedema associated with brimonidine therapy. PMID- 12648056 TI - Relationship between intraocular pressure and glaucomatous optic neuropathy. PMID- 12648058 TI - Palmodigital purpura as the only skin abnormality in myeloma-associated systemic amyloidosis. PMID- 12648059 TI - Extramedullary haemopoiesis in haemoglobin E/beta-thalassaemia. PMID- 12648060 TI - Glucocorticoid resistance in childhood leukaemia: mechanisms and modulation. PMID- 12648061 TI - Transient leukaemia--a benign form of leukaemia in newborn infants with trisomy 21. PMID- 12648062 TI - Di Guglielmo and his syndromes. PMID- 12648063 TI - Cytogenetics of multiple myeloma: interpretation of fluorescence in situ hybridization results. AB - The cytogenetic picture in multiple myeloma (MM) is highly complex, from which non-random numerical and structural chromosomal changes have been identified. Specifically, translocations involving the immunoglobulin heavy chain gene (IGH) at 14q32 and either monosomy or deletions of chromosome 13 have been reported in a significant number of patients from both cytogenetic and interphase fluorescence in situ hybridization (FISH) studies. Importantly, these abnormalities of chromosome 13 have recently been associated with a poor prognosis. In view of the highly complex nature of the karyotypes in MM patients, interphase FISH results may be difficult to interpret. In this study, cytogenetics and/or interphase FISH were carried out on bone marrow samples or purified plasma cells from 37 MM patients. Abnormal karyotypes, characterized by multiplex FISH (M-FISH) were found in 11 patients, all of which were highly complex. Interphase FISH revealed translocations involving the IGH locus in 16 (43%) patients. The IGH/cyclin D1 (CCND1) gene fusion characteristic of the translocation, t(11;14)(q13;q32), was seen in 12 (32%) of these patients and other rearrangements of IGH in four (11%) patients. Fourteen patients had additional copies of chromosome 11. Twenty patients (54%) had 13q14 deletions, 10 of whom also had t(11;14) or another IGH translocation. By comparing cytogenetic and FISH results, this study has revealed that significant chromosomal abnormalities might be hidden within highly complex karyotypes. Therefore, extreme caution is required in the interpretation of interphase FISH results in MM, particularly in relation to certain abnormalities, such as 13q14 deletions, which have an impact on prognosis. PMID- 12648064 TI - Soluble urokinase-type plasminogen activator receptor (suPAR) as an independent factor predicting worse prognosis and extra-bone marrow involvement in multiple myeloma patients. AB - The urokinase-type plasminogen activator (uPA) system, which consists of a proteinase (uPA), a receptor (uPAR or CD87) and inhibitors, is involved in proteolysis, cell migration, tissue remodelling, angiogenesis and cell adhesion. Recent findings suggest that malignant plasma cells express uPA and uPAR. The expression of these factors could represent a process by which myeloma plasma cells interact with the bone marrow (BM) environment and influence important biological events such as bone matrix degradation, plasma cell invasion and homing and, possibly, clinical evolution. We evaluated uPAR (CD87) and its soluble form (suPAR) in 49 multiple myeloma (MM) patients and correlated their expression and levels with clinico-biological characteristics of the disease. Flow cytometric analysis demonstrated that CD87 was expressed in all MM patients. High CD87 expression was associated with higher intensity of expression of CD56 (P = 0.038), CD38 (P = 0.058) and CD138 (P = 0.054) and CD45bright positivity (P = 0.014). suPAR levels correlated positively with soluble serum CD138 (P = 0.001), creatinine (P = 0.001), beta2-microglobulin (P < 0.001), disease stage (P = 0.017) and extra-BM involvement (P = 0.002). In the 46 evaluable patients, multivariate analysis showed that high levels of suPAR (P = 0.0214) and disease stage (P = 0.0064) were predictive of extra-BM involvement. In multivariate Cox analysis, 13q deletion (P = 0.0278), high soluble serum CD138 (P = 0.0201) and high suPAR (P = 0.0229) were the only parameters that independently affected survival. We conclude that CD87 is expressed on myeloma plasma cells and that suPAR, which predicts extra-BM involvement and poor prognosis, possibly represents a molecule with a relevant role in the biology of MM. PMID- 12648065 TI - A pooled analysis of karyotypic patterns, breakpoints and imbalances in 783 cytogenetically abnormal multiple myelomas reveals frequently involved chromosome segments as well as significant age- and sex-related differences. AB - The cytogenetic features (ploidy, complexity, breakpoints, imbalances) were ascertained in 783 abnormal multiple myeloma (MM) cases to identify frequently involved chromosomal regions as well as a possible impact of age/sex. The series included MM patients from the Mitelman Database of Chromosome Aberrations in Cancer and from our own laboratory. Hyperdiploidy was most common, followed by hypodiploidy, pseudodiploidy and tri-/tetraploidy. Most cases were complex, with a median of eight changes per patient. The distribution of modal numbers differed between younger and older patients, but was not related to sex. No sex- or age related differences regarding the number of anomalies were found. The most frequent genomic breakpoints were 14q32, 11q13, 1q10, 8q24, 1p11, 1q21, 22q11, 1p13, 1q11, 19q13, 1p22, 6q21 and 17p11. Breaks in 1p13, 6q21 and 11q13 were more common in the younger age group. The most frequent imbalances were + 9, - 13, + 15, + 19, + 11 and - Y. Trisomy 11 and monosomy 16 were more common among men, while -X was more frequent among women. Loss of Y as the sole change and + 5 were more common in elderly patients, and - 14 was more frequent in the younger age group. The present findings strongly suggest that some karyotypic features of MM are influenced by endogenous and/or exogenous factors. PMID- 12648066 TI - Gemcitabine, cisplatin and methylprednisolone chemotherapy (GEM-P) is an effective regimen in patients with poor prognostic primary progressive or multiply relapsed Hodgkin's and non-Hodgkin's lymphoma. AB - This study was designed to assess the efficacy and safety of gemcitabine, cisplatin and methylprednisolone (GEM-P) for patients with relapsed or refractory Hodgkin's disease (HD) and non-Hodgkin's lymphoma. Twenty-one patients were treated with gemcitabine (1000 mg/m2 d 1, 8 and 15), cisplatin (100 mg/m2 d 15) and methylprednisolone (1000 mg d 1-5) given every 28 d. Of these, 20 patients were evaluable for response. The median age was 38 years (range 17-64 years). Histological subtypes were: nodular sclerosing HD (n = 10), diffuse large B cell (n = 5), T cell-rich B cell (n = 2), follicular (n = 2), mantle cell (n = 1) and enteropathy-associated T-cell lymphoma (n = 1). The median remission duration prior to receiving GEM-P was only 42 d. The overall objective response rate was 80%[95% confidence interval (CI): 56-94%], including five complete and 11 partial responses. GEM-P induced responses in all histological subtypes, primary progressive disease and patients who had received a previous autograft. The only grade 3-4 toxicity was myelosuppression. However, no cases of febrile neutropenia or haemorrhage with thrombocytopenia were encountered. Median survival has not yet been reached and survival probability at 1 year was 60.8% (95% CI: 31.9 80.5%). In conclusion, GEM-P is a novel combination salvage therapy for poor prognostic primary progressive or multiply relapsed lymphoma patients. It has clinically significant activity with a favourable toxicity profile. PMID- 12648067 TI - Autologous stem cell transplant for relapsed and refractory peripheral T-cell lymphoma: variable outcome according to pathological subtype. AB - The purpose of this study was to evaluate the outcome of high-dose chemotherapy (HDCT) followed by autologous haematopoietic stem cell transplant (ASCT) for patients with relapsed T-cell non-Hodgkin's lymphoma. We reviewed 36 patients with peripheral T-cell lymphoma (PTL) who underwent ASCT between January 1987 and June 2001. Patients had chemosensitive disease, and received high-dose melphalan and etoposide with or without total body irradiation supported by unpurged autologous stem cells. Comparisons were made with 97 diffuse large B-cell lymphoma (DLBL) patients. PTL patients had a median age of 46 years (19-62 years). Twenty-nine had relapsed and seven had primary refractory disease. DLBL patients were statistically similar in baseline characteristics. Of patients with PTL, six (17%) died of treatment-related complications and 14 (39%) were in remission with a median follow-up of 42 months (range 6-116 months). Three-year overall survival and event-free survival (EFS) were 48% and 37%, respectively, for PTL, compared with 53% and 42% for DLBL (P = 0.41 and 0.29 respectively). There was no significant prognostic variable found by univariate analysis for the PTL cohort. Major PTL subtypes were analysed for outcomes. The 20 patients with PTL, not otherwise specified (PTL-NOS), had an inferior EFS compared with DLBL patients (23%, P = 0.028). In contrast, the nine patients with anaplastic large T/null cell lymphoma had a non-significant trend for improved EFS (67%, P = 0.41). While ASCT in patients with relapsed or primary refractory PTL results in long-term remission rates comparable to DLBL patients, those with PTL-NOS do significantly worse. PMID- 12648068 TI - Over-expression of short isoforms of Helios in patients with adult T-cell leukaemia/lymphoma. AB - In previous studies, we demonstrated an over-expression of the dominant-negative isoform of the transcription factor Ikaros in patients with blast crisis of both chronic myelogenous leukaemia and B-cell acute lymphoblastic leukaemia (ALL). Recently, we reported an over-expression of the short isoforms of Helios, which is one of the members of the Ikaros gene family, in a patient with T-cell ALL. In the present study, we found over-expression of short isoforms of Helios in human T lymphotropic virus-I (HTLV1)-infected patients who had developed chronic and acute forms of adult T-cell leukaemia/lymphoma. In contrast, we could not detect any over-expression of short isoforms of Helios in healthy HTLV1 carriers. By Southern blotting, we detected a small deletion in the Helios gene locus of adult T-cell leukaemia/lymphoma patients. The present results suggest that Helios gene abnormalities might be one of the important mechanisms in the disease progression of HTLV1 infection. PMID- 12648069 TI - The early molecular response to imatinib predicts cytogenetic and clinical outcome in chronic myeloid leukaemia. AB - Real-time quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR) is increasingly used to monitor responses in chronic myeloid leukaemia (CML). The peripheral blood BCR-ABL/ABL ratio, as assessed by Q-RT-PCR, has been shown to correlate with the contemporary cytogenetic response in patients receiving imatinib (Glivec, Gleevec). We have used Q-RT-PCR to monitor the early molecular response to 4 weeks and 3 months of imatinib therapy, in 47 patients with established CML. After 4 weeks of imatinib therapy, patients whose BCR-ABL/ABL ratio had fallen to less than 50% that of baseline had a significantly higher probability of achieving a major cytogenetic response after 6 months of therapy, when compared with those whose ratio did not fall by this amount (P < 0.001). Similarly, patients whose ratio at 3 months was less than 10% of that at baseline had a significantly higher probability of achieving a major cytogenetic remission at 6 months (P < 0.001). Patients who achieved these falls in their BCR-ABL/ABL ratio at either 4 weeks or 3 months had a superior progression-free survival at a median follow-up of 16.5 months (P = 0.01 and 0.003 respectively). These effects were independent of patient age and disease stage. The occurrence of peripheral blood cytopenias sufficiently severe to interrupt therapy was unrelated to progression-free survival. In conclusion, the data suggest that the early trend in the BCR-ABL/ABL ratio may be clinically useful for the early identification of patients destined to fare poorly on imatinib. PMID- 12648070 TI - Expression of human leucocyte antigens and co-stimulatory molecules on blasts of patients with acute myeloid leukaemia. AB - Recently, leukaemia-associated antigens (LAA) recognized by T lymphocytes, such as Wilm's tumour-1 (WT-1) or pathogenesis-related protein-1 (PR-1), have been identified. For immunotherapies that employ antigen peptides, either alone or pulsed on dendritic cells (DC), the expression of human leucocyte antigen (HLA) molecules on the targeted leukaemic blasts (LB) is crucial. The co-stimulatory molecules CD80 and CD86 give the secondary signal to T lymphocytes that is necessary for the lysis of leukaemia cells, and CD40 enhances the efficacy of antigen presentation. Here, the expression of HLA-ABC, HLA-A2, HLA-DR, CD40, CD80 and CD86 was flow cytometrically examined in blood samples from 24 healthy volunteers (HV), 24 patients with newly diagnosed acute myeloid leukaemia (AML) and five patients with relapsed AML. The expression of HLA-ABC, CD40, CD80 and CD86 was significantly reduced on LB in comparison with monocytes of HV. HLA-A2 and HLA-DR expression was similar on LB and on monocytes of HV. In AML patients, the expression of HLA and CD86 molecules was significantly higher on LB than on CD33/CD34-negative monocytes. CD40 and CD80 molecules were deficient on AML blasts. The preservation of HLA molecules and CD86 on LB of the majority of AML patients at the time of diagnosis and even at relapse of the disease are prerequisites for LAA-targeted immunotherapies in these patients. PMID- 12648071 TI - Resistance to spontaneous apoptosis in acute myeloid leukaemia blasts is associated with p-glycoprotein expression and function, but not with the presence of FLT3 internal tandem duplications. AB - The ability of acute myeloid leukaemia (AML) blasts to survive in culture has been associated with poor patient response to chemotherapy. Other biological factors predicting an adverse outcome include p-glycoprotein (pgp) expression, which is associated with a reduced remission rate, and the presence of fms-like tyrosine kinase 3 gene (FLT3) internal tandem duplications (ITDs), predictive of a high rate of leukaemic relapse. Our previous work has indicated a drug efflux independent role for pgp in apoptosis resistance. We measured spontaneous in vitro apoptosis in 58 primary AML samples to establish its relationship with functional and phenotypic pgp and with FLT3 ITDs. Cells were incubated for 48 h in a suspension culture, and the remaining viable cells were counted by flow cytometry. Median survival was 38% of baseline values. Resistance to spontaneous apoptosis was strongly associated with pgp (MRK-16 antibody) expression (P = 0.001) and with pgp functional activity (P < 0.001). FLT3 ITDs, found in 20 cases, were inversely associated with functional pgp activity: thus, the median pgp modulation ratio was 2.0 in FLT3 wild-type cases and 1.38 in ITD cases (P = 0.018). Also, the presence of FLT3 ITDs was not associated with in vitro apoptosis resistance. In conclusion, we have found that the presence of FLT3 ITDs is not related to AML blast survival in vitro, and is inversely associated with pgp activity, whereas pgp expression and activity are associated with resistance to spontaneous apoptosis. These results may help to explain the differing adverse effects of pgp (on remission induction) and FLT3 ITDs (on relapse) in AML. PMID- 12648072 TI - Prognostic significance of CD38 and CD20 expression as assessed by quantitative flow cytometry in chronic lymphocytic leukaemia. AB - CD38 expression on chronic lymphocytic leukaemia (CLL) cells is a poor prognostic factor, however, methods for measuring this vary. The QuantiBRITETM flow cytometry (FC) system yields an absolute antigen expression value (antibodies bound/cell, ABC) and may be useful in standardizing CD38 expression analysis. We evaluated cryopreserved pretreatment CLL cells for CD20 ABC, CD38 ABC, and percentage of CD38+ B cells from 131 patients requiring therapy. The 92 patients (70%) with >/= 100 CD38 ABC had worse overall survival (OS; 34% at 5 years) compared with those with < 100 CD38 ABC (70% at 5 years, mortality hazard ratio 2.30, 95% confidence interval 1.28-4.12; two-tailed P = 0.003). Among the 64 patients with < 30% CD38+ cells, OS of the 25 with >/= 100 ABC was worse than that of the 39 with < 100 ABC (P = 0.018). OS of patients with < 30% CD38+ cells and >/= 100 ABC was actually similar to that of patients with >/= 30% CD38+ cells. BrightCD20 expression (>/= 20 000 ABC) was not associated with a worse OS (P = 0.10). The presence of >/= 100 CD38 ABC in CLL patients requiring therapy is an unfavourable prognostic factor for OS and quantitative FC may be superior to percentage CD38+ cell assessment. Prospective trials are required to determine more precisely the prognostic significance of absolute expression levels in fresh CLL cells. PMID- 12648073 TI - Demonstration of aberrant T-cell and natural killer-cell antigen expression in all cases of granular lymphocytic leukaemia. AB - The diagnosis of granular lymphocytic leukaemia (GLL) requires the presence of an immunophenotypically distinct T-cell (T-GLL) or natural killer-cell (NK-GLL) population. Flow cytometric immunophenotyping was performed on 21 T-GLL patients, 11 NK-GLL patients and 20 normal control subjects using antibodies to T and NK cell-associated antigens in order to accurately identify the distinguishing features of T-GLL and NK-GLL. The NK antigens evaluated included: CD16, CD57, CD94, CD161, and the killing inhibitory receptors (KIRs) CD158a, CD158b and CD158e (p70). Abnormal T-antigen expression was present in all T-GLL patients. CD57 was frequently expressed in T-GLL, however, one-third of patients showed partial CD57 expression similar to that seen in T cells from normal control subjects. Ten T-GLL were KIR positive; all expressed a single KIR isoform. All NK GLL showed a distinctive, abnormal immunophenotype. Four NK-GLL expressed a single KIR isoform; the remaining seven patients lacked all tested KIRs, which is also a distinct, abnormal finding. Immunoperoxidase staining of bone marrow biopsy specimens from NK-GLL patients with antibodies to CD8, TIA-1 and granzyme B revealed the disease-specific distinctive staining patterns previously found in T-GLL. These studies delineate the unique immunophenotypic features diagnostic of T-GLL and provide strong evidence that NK-GLL, like T-GLL, represents a clonal lymphoproliferative disorder. PMID- 12648074 TI - A validated decision model for treating the anaemia of myelodysplastic syndromes with erythropoietin + granulocyte colony-stimulating factor: significant effects on quality of life. AB - We have published previously a prototype of a decision model for anaemic patients with myelodysplastic syndromes (MDS), in which transfusion need and serum erythropoietin (S-Epo) were used to define three groups with different probabilities of erythroid response to treatment with granulocyte colony stimulating factor (G-CSF) + Epo. S-Epo 500 U/l and >/= 2 units/month for a poor response, whereas the presence of only one negative prognostic marker predicted an intermediate response. A total of 53 patients from a prospective study were included in our evaluation sample. Patients with good or intermediate probability of response were treated with G CSF + Epo. The overall response rate was 42% with 28.3% achieving a complete and 13.2% a partial response to treatment. The response rates were 61% and 14% in the good and intermediate predictive groups respectively. The model retained a significant predictive value in the evaluation sample (P < 0.001). Median duration of response was 23 months. Scores for global health and quality of life (QOL) were significantly lower in MDS patients than in a reference population, and fatigue and dyspnoea was significantly more prominent. Global QOL improved in patients responding to treatment (P = 0.01). The validated decision model defined a subgroup of patients with a response rate of 61% (95% confidence interval 48 74%) to treatment with G-CSF + Epo. The majority of these patients have shown complete and durable responses. PMID- 12648075 TI - Infection--an underappreciated cause of bone pain in multiple myeloma. AB - Bone pain, especially back pain, is a common presenting feature of myeloma patients. We report three multiple myeloma patients with exacerbations of back pain and referred shoulder pain resulting from vertebral infections. Two patients were treated with surgery, and one patient had computerized tomography-guided percutaneous needle aspiration for diagnostic purposes. All three patients received a prolonged course of antibiotics. Vertebral infection resolved with this treatment in all three patients without any recurrence. Previous dexamethasone therapy, together with an episode of bacteraemia, appears to be a predisposing factor for vertebral infection. Magnetic resonance imaging enabled the diagnosis in all three patients. PMID- 12648076 TI - Methionine synthase genetic polymorphism MS A2756G alters susceptibility to follicular but not diffuse large B-cell non-Hodgkin's lymphoma or multiple myeloma. AB - Lymphoproliferative diseases are characterized by chromosomal aberrations, and susceptibility may depend on inherited activity of enzymes required for DNA synthesis and methylation. We analysed genetic polymorphisms for methionine synthase (MS) A2756G, methylenetetrahydrofolate reductase (MTHFR) C677T and MTHFR A1298C in Caucasians with non-Hodgkin's lymphoma (NHL; n = 151), multiple myeloma (MM; n = 90) and 299 control subjects. The MS 2756 AG/GG genotypes were significantly under-represented in NHL (26.2%) vs control subjects (37.2%; P = 0.02), and conferred a 2.4-fold lower risk of follicular (odds ratio = 0.41, 95% confidence interval: 0.19-0.88, p = 0.02) but not diffuse large B-cell lymphoma. MM patients showed no significant difference in the polymorphisms compared with control subjects. PMID- 12648077 TI - High levels of hyaluronan production by a malignant lymphoma cell line with primary effusion lymphoma immunophenotype OHK. AB - We investigated whether hyaluronan (HA) was produced by a malignant lymphoma cell line with a primary effusion lymphoma immunophenotype, OHK. OHK secreted high levels of HA into the culture supernatant, and reverse transcription polymerase chain reaction showed mRNA for the HA synthases 2 and 3. HA binding protein reactive HA was present in the plasma membrane and in the cytoplasm. The inoculation of OHK into severe combined immunodeficiency disease mice produced marked viscid ascites with concentrated HA and a vascular-rich mesenteric solid tumour. These results show that HA is produced and secreted by the malignant lymphoma cell line OHK. PMID- 12648078 TI - Leukaemic pulmonary infiltrates in adult acute myeloid leukaemia: a high resolution computerized tomography study. AB - Leukaemic infiltration of the lungs may occur in acute myeloid leukaemia (AML). Pulmonary infiltrates are usually microscopic and invariably associated with hyperleucocytosis. Four AML patients with respiratory symptoms and low leucocyte counts underwent standard chest radiography, bronchoscopy with bronchoalveolar lavage and high-resolution computerized tomography (HRCT) of the lungs. HRCT scans showed pulmonary infiltrates with alveolar, interstitial, mixed and peribronchial/perivascular patterns in all patients, including one with negative standard radiographic findings. Infectious agents were excluded. Histology of the lung biopsy/autopsy specimens showed leukaemic infiltrates. Pulmonary leukaemia may be the cause of pulmonary infiltrates, even in non-hyperleucocytosic AML patients with low blast counts. PMID- 12648080 TI - Lack of association of beta2-glycoprotein I polymorphisms Val247Leu and Trp316Ser with antiphospholipid antibodies in patients with thrombosis and pregnancy complications. AB - Beta2-glycoprotein I (beta2GPI) is an important target antigen for antiphospholipid antibodies (aPL) and thus beta2GPI polymorphisms may influence aPL production and the development of antiphospholipid syndrome. We have studied the relationship between the Val247Leu and Trp316Ser beta2GPI polymorphisms and the aPL status of 230 patients referred for aPL screening. Sixty-one (26.5%) had persistent aPL [anticardiolipin antibodies (IgG and/or IgM), lupus anticoagulants and/or IgG anti-beta2GPI antibodies]. A comparison of the genotypic and allelic frequencies of these two polymorphisms between the Caucasian patient population and an ethnic-matched normal control group (n = 308) showed no significant differences between aPL-positive patients, aPL-negative patients and the normal control group. This suggests that the Val or Leu allele at position 247 and the Trp or Ser allele at position 316 of beta2GPI do not play a role in the production of aPL. There was a significantly decreased prevalence of the Ser316 allele in aPL-negative women (n = 98) when compared with female normal control subjects (n = 249) [0.020 [95% confidence interval (CI) 0.00-0.04]vs 0.060 (95% CI 0.04-0.08), P = 0.0286]. Subgroup analysis showed no significant difference between female patients with thrombosis and female normal control subjects. Thus, the Ser316 allele may protect women from developing pregnancy complications by influencing an anticoagulant function of beta2GPI via a mechanism distinct from aPL production. PMID- 12648079 TI - Aberrant p15 gene promoter methylation in therapy-related myelodysplastic syndrome and acute myeloid leukaemia: clinicopathological and karyotypic associations. AB - Seventeen patients with therapy-related myelodysplastic syndrome/acute myeloid leukaemia (t-MDS/AML) were examined for aberrant p15 gene methylation by methylation-specific polymerase chain reaction. Ten patients (58%) showed p15 methylation, which was significantly related to monosomy/deletion of chromosome 7q, but not to antecedent chemotherapy, blast count, leukaemic evolution or survival. In three of six patients with marrow samples obtained prior to the diagnosis of t-MDS/AML, p15 methylation predated disease development by up to 2 years. Bone marrow transplantation led to the disappearance of p15 methylation in one patient. These results showed that p15 methylation was an early event in the evolution of some t-MDS/AML patients. PMID- 12648081 TI - Ultrasound screening for internal jugular vein thrombosis aids the detection of central venous catheter-related infections in patients with haemato-oncological diseases: a prospective observational study. AB - To prove the hypothesis that central venous catheter-related thrombosis and infection are associated, 43 haemato-oncological patients with an internal jugular vein catheter underwent ultrasound screening for thrombosis every 4 d. Catheter-related thrombosis was detected in 13/43 patients (30%). Catheter related infection, as defined by the U.S. Hospital Infection Control Practices Advisory Committee, was found in 14/43 patients (33%) with colonization of the catheter in two patients, exit site infection in eight patients and catheter related bloodstream infection in four patients. Catheter-related thrombosis and catheter-related infection coincided in 12 patients and were significantly correlated (Fisher's exact test, P < 0.0001). Detection of thrombosis indicated a catheter-related infection with a superior sensitivity (86% vs 57%) and an equivalent specificity (97%) compared with the presence of clinical signs (erythema, tenderness, warmth or swelling). Neutropenia, which occurred in 32 patients, was found in 13/14 patients (93%) with a catheter-related infection and, therefore, seemed to be an important covariate for the development of a catheter-related infection. This study showed a close correlation between catheter-related thrombosis and infection. Ultrasound screening for thrombosis was helpful for detecting catheter-related infection. These findings could be clinically useful for the handling of central venous catheters in patients with an elevated risk of infectious complications. PMID- 12648082 TI - Long-term follow-up of chronic autoimmune thrombocytopenic purpura refractory to splenectomy: a prospective analysis. AB - Splenectomy remains the most effective treatment of chronic autoimmune idiopathic thrombocytopenia (ITP) (i.e. of > 6 months duration). Treatment of patients refractory to splenectomy (with absence of response or relapse after initial response) is difficult, and their long-term outcome is not well known. Over a 10 year period, 183 patients with chronic ITP were splenectomized including 158 adults and 25 children ( 100 x 10(9)/l, nine of them without treatment and 27 of them with low-dose steroids or azathioprine; six (13%) remained moderately thrombocytopenic (35 x 10(9)/l to 100 x 10(9)/l platelets); the last five patients, without response to any treatment (up to six regimens), remained severely thrombocytopenic (platelets < 20 x 10(9)/l), and three of them died from bleeding. Twenty-seven (57%) of the 47 refractory cases required at least one hospitalization, in the majority of cases for intravenous immunoglobulin (IVIg) infusions. Seven of the refractory cases occurred in children. Six of them subsequently reached platelet counts > 100 x 10(9)/l, but one died from bleeding. Our findings confirm the overall favourable long-term prognosis of chronic ITP refractory to splenectomy. PMID- 12648083 TI - Screening strategies in C282Y-linked haemochromatosis. PMID- 12648084 TI - The dangers of iron overload in pyruvate kinase deficiency. PMID- 12648085 TI - Photosensitization in chronic myelogenous leukaemia patients treated with imatinib mesylate. PMID- 12648086 TI - Successful combination of anti-CD33 antibody (gemtuzumab ozogamicin) and minimal conditioning before second allografting in recurrent acute myeloid leukaemia. PMID- 12648087 TI - Methylenetetrahydrofolate reductase 677CC normal genotype may protect against multiple myeloma. PMID- 12648090 TI - Origin of rabbit (Oryctolagus cuniculus) in China: evidence from mitochondrial DNA control region sequence analysis. AB - A fragment of mitochondrial DNA (mtDNA) control region (approximately 700 bp) was sequenced in 104 individuals from 20 breeds (three Chinese domestic breeds, five recently derived breeds and 12 introduced breeds) of domestic rabbits, Oryctolagus cuniculus. Nineteen sites were polymorphic, with 18 transitions and one insertion/deletion, and eight haplotypes (A1, A2, A3, A4, A5, A6, A7 and A8) were identified. Haplotype A1 was the most common and occurred in 89 individuals. In the 25 Chinese rabbits, only haplotype A1 was observed, while four haplotypes (A1, A3, A5 and A6) were found in 26 recently derived individuals. Haplotype A2 was shared by seven individuals among three introduced strains. The other six haplotypes accounted for 0.96-1.92% of the animals. Combined with the published sequences of European rabbits, a reduced median-joining network was constructed. The Chinese rabbit mtDNAs were scattered into two clusters of European rabbits. These results suggest that the (so-called) Chinese rabbits were introduced from Europe. Genetic diversity in Chinese rabbits was very low. PMID- 12648091 TI - Genotypic variability at the major histocompatibility complex (B and Rfp-Y) in Camperos broiler chickens. AB - Evidence for the importance of major histocompatibility complex (MHC) genotype in immunological fitness of chickens continues to accumulate. The MHC B haplotypes contribute resistance to Marek's and other diseases of economic importance. The Rfp-Y, a second cluster of MHC genes in the chicken, may also contribute to disease resistance. Nevertheless, the MHC B and Rfp-Y haplotypes segregating in broiler chickens are poorly documented. The Camperos, free-range broiler chickens developed in Argentina, provide an opportunity to evaluate MHC diversity in a genetically diverse broiler stock. Camperos are derived by cross-breeding parental stocks maintained essentially without selection since their founding. We analysed 51 DNA samples from the Camperos and their parental lines for MHC B and Rfp-Y variability by restriction fragment pattern (rfp) and SSCP typing methods for B-G, B-F (class Ia), B-Lbeta (class II) and Y-F (class Ib) diversity. We found evidence for 38 B-G genotypes. The Camperos B-G patterns were not shared with White Leghorn controls, nor were any of a limited number of Camperos B-G gene sequences identical to published B-G sequences. The SSCP assays provided evidence for the presence of at least 28 B-F and 29 B-Lbeta genotypes. When considered together B-F, B-L, and B-G patterns provide evidence for 40 Camperos B genotypes. We found even greater Rfp-Y diversity. The Rfp-Y class I-specific probe, 163/164f, revealed 44 different rfps among the 51 samples. We conclude that substantial MHC B and Rfp-Y diversity exists within broiler chickens that might be drawn upon in selecting for desirable immunological traits. PMID- 12648092 TI - Phylogenies using mtDNA and SRY provide evidence for male-mediated introgression in Asian domestic cattle. AB - Using nucleotide sequences of the mitochondrial DNA (mtDNA) cytochrome b and SRY genes, we examined the genetic status of two major groups of domestic cattle, the humpless taurine (Bos taurus) and humped zebu (B. indicus), using 10 cattle populations in Asia. Several sequence polymorphisms specific for each major group were found, although the frequency of these polymorphisms varied in each population. Six major mtDNA-SRY composite types were observed. The Mishima, Mongolian, Korean, Chinese Yellow and Sri Lanka cattle populations had a full match between the mtDNA and SRY sequences, specifically the taurine/taurine type or zebu/zebu type. A non-match type (zebu/taurine type) was found at a high frequency in the Bangladesh (83.4%) and Nepal populations (83.3%). Our results suggest that these non-match type populations developed from genetic hybridization of different strains. Also, the domestication history of modern Asian domestic cattle could be explained by male-mediated introgression. Additionally, our results suggest the occurrence of introgression of mtDNA from other Bibos or Poephagus species into native cattle populations. The existence of other mtDNA-SRY composite types, such as the Bali-zebu and yak-zebu types in Indonesia (85.7%) and Nepal (16.7%), respectively, suggests that genetic introgression also occurred from other genera into domestic cattle during the process of domestication. PMID- 12648093 TI - A consolidated linkage map for rainbow trout (Oncorhynchus mykiss). AB - Androgenetic doubled haploid progeny produced from a cross between the Oregon State University and Arlee clonal rainbow trout (Oncorhynchus mykiss) lines, used for a previous published rainbow trout map, were used to update the map with the addition of more amplified fragment length polymorphic (AFLP) markers, microsatellites, type I and allozyme markers. We have added more than 900 markers, bringing the total number to 1359 genetic markers and the sex phenotype including 799 EcoRI AFLPs, 174 PstI AFLPs, 226 microsatellites, 72 VNTR, 38 SINE markers, 29 known genes, 12 minisatellites, five RAPDs, and four allozymes. Thirty major linkage groups were identified. Synteny of linkage groups in our map with the outcrossed microsatellite map has been established for all except one linkage group in this doubled haploid cross. Putative homeologous relationships among linkage groups, resulting from the autotetraploid nature of the salmonid genome, have been revealed based on the placement of duplicated microsatellites and type I loci. PMID- 12648094 TI - Characterization of naturally processed and presented peptides associated with bovine major histocompatibility complex (BoLA) class II DR molecules. AB - Differential regulation of genetic resistance to infectious disease may partially be explained by variation in the binding affinity and the repertoire of pathogen derived antigenic peptides associated with major histocompatibility complex (MHC) molecules. In this study, we investigated characteristics of peptides that bind to the bovine MHC allele BoLA-DRB3*2703, which is associated with occurrence of clinical mastitis in Holstein dairy cattle, and assigned a putative peptide binding motif to this allele. This was achieved by in vitro expression of allele *2703 as well as a control allele, BoLA-DRB3*1201 which is present at high frequency in Holsteins. Transfected cell lines alone (for allele *1201) or in combination with blood mononuclear cells from an animal homozygous for allele *2703 were used as the source of naturally processed and presented peptides. Subsequent to elution of peptides from BoLA-DR+ cells, their sequences were determined by electrospray ionization mass spectrometry. Eluted peptides were between 13 and 20 amino acids long and the majority were in sets of overlapping sequences. These peptides were derived from intra- and extracellular proteins, as well as foreign proteins present in the culture medium. Some peptides had originated from molecular chaperones present in the endoplasmic reticulum, such as ER-60 and GRP78, pointing to some degree of overlap and cross-sampling between MHC class I and class II antigen presentation pathways. Consistent with reports of human and mouse MHC class II-associated peptides, putative peptide-binding motifs could be assigned to alleles *2703 and *1201, comprising a hydrophobic or an aromatic residue at relative position 1, a hydrophobic residue at position 4 and a small residue at position 6 of the eluted peptides. These findings provide the foundation for future studies of molecular mechanisms of MHC-disease associations of cattle. PMID- 12648095 TI - Cloning and characterization of the bovine EGR4 gene and evaluation as candidate gene for bovine spinal dysmyelination. AB - Genes of the early growth response (EGR) family encode transcription factors with a highly conserved DNA binding zinc finger domain, which regulate a variety of genes, e.g. late myelin genes. Here, the cloning, genomic structure and expression of the bovine orthologue of the EGR4 gene are reported. The gene consists of two exons and encodes a 482 amino acid protein with a Cys2His2 zinc finger structure. The predicted protein shares between 80 and 87% identity to mouse, rat and human EGR4 proteins and all four species share almost complete identity in the DNA-binding domain. The bovine transcript is alternatively spliced by retaining intronic sequence, giving rise to two different mRNAs differing in three nucleotides and resulting in an extra alanine residue in the longer variant of the predicted protein. The gene was mapped by radiation hybrid (RH) mapping to markers on bovine chromosome 11. EGR4 transcripts were detected by reverse transcriptase polymerase chain reaction (RT-PCR) in the frontal cortex and cerebellum, and a low expression level was also detected in the liver. The EGR4 gene was evaluated as a candidate gene for bovine spinal dysmyelination (BSD). Sequencing of the gene from a homozygous affected animal and a heterozygous carrier revealed a single base mutation that leads to an amino acid substitution at residue 322 in EGR4. Genotype analysis of this polymorphism in a pedigree segregating for BSD, as well as in a panel of different cattle breeds, and sequence analysis of the entire coding region suggested that the EGR4 is not the gene responsible for BSD. Furthermore, 87 animals of different cattle breeds were screened for single-nucleotide polymorphisms (SNPs) resulting in the identification of two SNPs in EGR4. PMID- 12648096 TI - Mapping of porcine ESTs obtained from the anterior pituitary. AB - We report the physical mapping of porcine expressed sequence tags (ESTs) from anterior pituitary clones isolated by differential display PCR in a study using lines selected for reproduction. These ESTs were mapped using a somatic cell hybrid panel (SCHP) and a radiation hybrid panel (IMpRH) as follows (SCHP position, nearest marker on the RH map): SPARCL1 (8q23-q27, SSP1); ATF4 (5p11 p15, AC02); MEF2C [2(1/2q21)-(1/2q22), SW2134]; FTH1 (2p14-p17, SWR783); FRAP1 (6q22-q23, SW1355); PBP (14, SW2508); LOC92004 [13q23-(1/2q41), CP]; and PGRMC1 [Xq22, SW1943]. All RH assignments were at LOD score >6.0 except for PGRMC1 at LOD score 5.4. ESTs TCP1 [12p11-(2/3p13)], SF3B1 (15q23-q26) and Clock (8q11-q12) were assigned using only the SCHP. The map position of SPARCL1 coincides with a quantitative trait loci (QTL) for age at puberty found in the University of Nebraska selection lines. Physical mapping of ESTs reported in the present study contributes to characterization of the transcriptome of anterior pituitary of pigs, adds new information to the public database of the porcine genome expression map, and further develops the porcine-human comparative map. PMID- 12648098 TI - Evaluation of the present breeding programme against copper toxicosis in Danish Bedlington terriers. AB - A breeding programme to eradicate copper toxicosis in Danish Bedlington terriers has been established based on a DNA marker test. Genotyping of both parents is compulsory and after 1 January 2000, only homozygous non-carriers are used for breeding. In this study, two groups of Bedlington terriers were genotyped at 18 microsatellite loci. One group represented the original population of Bedlington terriers before introducing the breeding programme (n = 23); the other represented a group of homozygous non-carriers (n = 24) available for breeding after year 2000. Allele numbers, allele frequencies, observed heterozygosities (Ho), expected heterozygosities (He), locus-specific coefficients of inbreeding (Fl) and Nei's genetic distance (D) was calculated. Individual coefficients of inbreeding (Fi) were calculated from the pedigrees and an assignment test was performed. Four rare alleles were lost in the group of homozygous non-carriers. No significant differences were observed between the mean values of allele numbers, Ho, He, Fl and Fi of the two populations of dogs. Nei's genetic distance between the two populations was 0.06 and 88% of the homozygous non-carriers were assigned correctly in the assignment test. The overall diversity of the breed was low (Ho = 0.41) and the breeders were advised to include the heterozygous carriers again. PMID- 12648097 TI - Development of 50 gene-associated microsatellite markers using BAC clones and the construction of a linkage map of swine chromosome 4. AB - The development of informative polymorphic markers is essential for QTL mapping. We developed 50 microsatellite markers from BAC clones containing genes that were predicted to map swine chromosome 4 (SSC4) according to comparative analysis between human and swine chromosomes, and constructed a linkage map that consisted of 37 markers including 24 markers closely linked to genes in BAC clones. Microsatellite markers were developed by direct-sequencing of BAC clones and our results demonstrated that this method was effective for developing microsatellite markers in specific regions on chromosomes. Effective development of microsatellite markers closely linked to genes can further accelerate the comparative studies of chromosomes between different species. PMID- 12648099 TI - Characterization of a SINE indel polymorphism in the porcine AGL gene and assignment of the gene to chromosome 4q. PMID- 12648100 TI - Polymorphisms in the equine WNT1 gene allow linkage mapping to ECA6q. PMID- 12648101 TI - Linkage mapping of POMC to bovine chromosome 11. PMID- 12648102 TI - Linkage mapping of FBN1 to bovine chromosome 10. PMID- 12648103 TI - Construction and characterization of three BAC libraries for analysis of the chicken genome. PMID- 12648105 TI - Characterization and mapping of 19 polymorphic microsatellite markers for rainbow trout (Oncorhynchus mykiss). PMID- 12648104 TI - Linkage mapping of the porcine testis enhanced gene transcript (TEGT) gene to chromosome 5. PMID- 12648106 TI - OtY1 is a Y-linked marker in Chinook salmon but not in Rainbow trout. PMID- 12648107 TI - Marker polymorphisms in the porcine genes for muscle glycogen synthase (GYS1) and muscle glycogen phosphorylase (PYGM). PMID- 12648108 TI - Genetic mapping of the chicken stem cell antigen 2 (SCA2) gene to chromosome 2 via PCR primer mutagenesis. PMID- 12648109 TI - Anaesthetists and care of the critically ill child. PMID- 12648111 TI - The effects of sevoflurane and desflurane in vitro on platelet-leukocyte adhesion in whole blood. AB - The interaction between platelets and leukocytes plays an important role in inflammatory and thrombotic processes. We investigated whether the volatile anaesthetics sevoflurane and desflurane alter the formation of platelet-leukocyte aggregates and the expression of P-selectin on platelets. Whole blood was incubated with 1 and 2 minimum alveolar concentration (MAC) sevoflurane or desflurane. Unstimulated and adenosine diphosphate, or thrombin receptor agonist peptide-6-stimulated samples were stained with fluorochrome-conjugated antibodies. The formation of platelet-leukocyte conjugates and the expression of P-selectin on platelets were measured using flow cytometry. Sevoflurane was found to enhance the binding of platelets to lymphocytes, neutrophils and monocytes, it also increased the expression of P-selectin on platelets especially in the stimulated samples. Desflurane decreased the percentage of lymphocyte-platelet, neutrophil-platelet and monocyte-platelet conjugates principally in unstimulated samples. The results show that these two volatile anaesthetics have differing effects on the formation of platelet-leukocyte conjugates in vitro. Sevoflurane also enhanced the expression of P-selectin on platelets. PMID- 12648112 TI - Mathematical modelling and simulation for planning critical care capacity. AB - Using average number of patients expected in a year, average length of stay and a target occupancy level to calculate the number of critical care beds needed is mathematically incorrect because of nonlinearity and variability in the factors that control length of stay. For a target occupancy in excess of 80%, this simple calculation will typically underestimate the number of beds required. More seriously, it provides no quantitative guidance information about other aspects of critical care demand such as the numbers of emergency patients transferred, deferral rates for elective patients and overall utilisation. The combination of appropriately analysing raw data and detailed mathematical modelling provides a much better method for estimating numbers of beds required. We describe this modelling approach together with evidence of its performance. PMID- 12648113 TI - Impact of an outreach team on re-admissions to a critical care unit. AB - Re-admissions have been cited as a measure of critical care quality and outreach teams have recently been introduced to improve critical care delivery. The aim of this study was to examine whether the number, causes and sequence of re admissions to critical care altered as a result of the introduction of an outreach team. Re-admissions between April 2000 and November 2001 were examined. The reasons for re-admission were classified as (i) same pathology or disease process; (ii) new, but related, pathology; (iii) new and unrelated pathology; (iv) exacerbation of other comorbidities. During the two-year period, a total of 2546 patients were admitted to critical care of which 100 were re-admitted (49 before outreach and 51 after outreach). The reasons for re-admission did not vary before or after the introduction of the outreach team (same pathology 15 vs. 15; new, but related, pathology 17 vs. 23; new, but unrelated, 14 vs. 9; exacerbation of comorbidity 3 vs. 4, respectively, Chi-squared = 2.07, df = 3, p = 0.56). There was also no difference between the duration of stay on the general ward in between the critical care unit admissions before (median 2.93 [interquartile range 1.32-6.05] days) or after (median 2.25 [interquartile range 1.06-6.32] days) the introduction of an outreach team. As we could not detect any change in patterns of re-admissions as a result of the introduction of an outreach team, we would suggest that although outreach is an important development for critical care, its performance should be measured by other parameters. PMID- 12648114 TI - A randomised crossover comparison of patient-controlled sedation and patient maintained sedation using propofol. AB - This randomised, crossover study compared patient-controlled sedation using boluses of propofol and patient-maintained sedation using a target-controlled infusion of propofol. Twenty-three patients aged 18-35 years having surgical removal of bilateral third molar teeth under local anaesthesia during two separate visits were studied. In the majority of patients, both techniques provided moderate sedation, good operating conditions, stable physiological parameters and a high degree of patient satisfaction. Two patients became over sedated during patient-controlled sedation. The time taken for titration to adequate sedation was longer with patient-maintained sedation than with patient controlled sedation [mean (SD) = 8.6 (3.7) min vs. 5.7 (3.1) min, p < 0.005]. The mean overall propofol consumption was similar with both techniques. The majority of patients preferred patient-maintained sedation to patient-controlled sedation, p < 0.05. PMID- 12648115 TI - Job satisfaction, stress and burnout in Australian specialist anaesthetists. AB - A postal survey was sent to specialist anaesthetists in Australia looking at aspects of job satisfaction, dissatisfaction and stress. Burnout was measured using the Maslach Burnout Inventory. The response rate was 60% (422/700) with the majority of respondents being male (83%). Stressful aspects of anaesthesia included time constraints and interference with home life. Experienced assistants and improved work organisation helped to reduce stress. The high standard of practice and practical aspects of the job were deemed satisfying, whereas poor recognition and long hours were the major dissatisfying aspects of the job. With respect to burnout, high emotional exhaustion, high levels of depersonalisation and low levels of personal achievement were seen in 20, 20 and 36% of respondents, respectively. Female anaesthetists reported higher stress levels than males (p = 0.006), but tended to prioritise home/work commitments better than males (p = 0.05). Private practitioners rated time issues of high importance compared with public hospital doctors, whereas public hospital doctors rated communication problems as being more significant than with private specialists. Although burnout levels are high in anaesthetists, they compare favourably with other medical groups. There are, however, aspects of the anaesthetist's job that warrant further attention to improve job satisfaction and stress. PMID- 12648116 TI - Eliminating protein from reusable laryngeal mask airways. A study comparing routinely cleaned masks with three alternative cleaning methods. AB - Laryngeal mask airways (LMAs) have the potential to act as a vector for the transmission of prion diseases. This study was undertaken to define the problem of protein contamination and to investigate three alternative cleaning methods. Forty-eight LMAs were allocated to one of four groups, stained with erythrosin and given a total stain score and a grid stain score in order to determine the degree of protein contamination. Eighteen randomly selected LMAs that had been routinely cleaned and sterilised (group 1) were compared with 12 LMAs that had been washed and scrubbed with the benefit of prior staining (group 2), 13 that had been washed and scrubbed without any visual guide (group 3) and 13 that had been cleaned using a Biosonic ultrasonic cleaning system (group 4). The results show that none of the cleaning methods achieved optimal results, as all methods left proteinaceous material on some masks. The study showed that: (i) staining as a guide to cleaning does not lead to more effective removal of proteinaceous material; (ii) systematic cleaning and scrubbing does lead to more effective removal of proteinaceous material from surfaces other than the grid area; and (iii) ultrasonic cleaning was more effective than other methods of cleaning for the removal of proteinaceous material from those areas of the mask most inaccessible, such as the grid. PMID- 12648117 TI - Mobile phones in the hospital--past, present and future. AB - The phenomenon of electromagnetic interference by mobile phones is real and potentially clinically significant. This has been recognised by the Department of Health and the Medical Devices Agency, leading to bans on phone use in hospitals. Current evidence suggests that mobile phones can cause malfunction of medical equipment, but only when used in close proximity. Allowing phone use in non patient care areas and improving staff education may improve compliance with hospital policies. PMID- 12648119 TI - Problems with disposable laryngoscope blades. PMID- 12648118 TI - The use of remifentanil in the anaesthetic management of patients undergoing adrenalectomy: a report of three cases. AB - The use of remifentanil has been recommended because of its ability to minimise the hypertensive response to tracheal intubation and surgical stimulation in various types of surgery. We describe the use of remifentanil in the anaesthetic management of three cases of open adrenalectomy, two for removal of a phaeochromocytoma and one for removal of an adrenal cortical tumour. Although the use of remifentanil was associated with no adverse events in the patient undergoing resection of the adrenal cortical tumour, its administration was associated with significant hypotension and bradycardia in the two phaeochromocytoma patients, who had both been given alpha- and beta-adrenergic receptor blocking drugs before surgery. It did not prevent the increases in blood pressure or plasma catecholamine levels associated with tumour manipulation in these patients. Remifentanil should therefore be used with caution in patients receiving alpha- and beta-adrenergic receptor blocking drugs. The use of potent vasodilators may still be necessary during tumour manipulation even if remifentanil is being infused. PMID- 12648120 TI - New GMC guidelines. PMID- 12648121 TI - Chinese herbal medicines. PMID- 12648122 TI - Herbal medicine and anaesthesia. PMID- 12648123 TI - Time constant or half time of a breathing system? PMID- 12648124 TI - Magnetic resonance compatible equipment. PMID- 12648125 TI - Thirteen centimetre central venous catheters, lucky for all? PMID- 12648126 TI - Critical incident involving an adjustable tracheostomy tube. PMID- 12648127 TI - Oxygen failure alarms. PMID- 12648129 TI - Laryngoceles and the laryngeal tube. PMID- 12648128 TI - Lingual tonsil hypertrophy with difficult airway and uncontrollable bleeding. PMID- 12648130 TI - Gum elastic bougie and simulated difficult intubation. PMID- 12648131 TI - Unexpected difficult airway. PMID- 12648132 TI - Time to switch to disposable bougies. PMID- 12648133 TI - Use of the laryngeal tube during cardiopulmonary resuscitation by paramedical staff. PMID- 12648134 TI - Failure of prefilled propofol syringe. PMID- 12648135 TI - Operative ascitic drainage in a patient with primary intestinal lymphangiectasia. PMID- 12648136 TI - An unusual anaesthetic for a through-knee amputation. PMID- 12648138 TI - Post dural puncture headache. PMID- 12648139 TI - False dermatome testing. PMID- 12648140 TI - An unusual cause of postpartum collapse or a red herring? PMID- 12648141 TI - Uncommon cause of itchy back in pregnancy. PMID- 12648142 TI - Blocked epidural catheter: time to look beyond the catheter. PMID- 12648143 TI - Teeth jewelry. PMID- 12648145 TI - Chancre very much! PMID- 12648146 TI - Mind your language. A response to 'Can we be more polite?', Shanker R, Anaesthesia 2002; 57: 1044. PMID- 12648147 TI - A response to 'A simple way to reduce heat loss from the head', Parry NS, Nightingale A, Anaesthesia 2002; 57: 1043. PMID- 12648148 TI - A response to 'Herbal medicines - harmless or harmful?', Sehgal A, Hall JE, Anaesthesia 2002; 57: 947-8. PMID- 12648149 TI - A response to 'A study of the initial fluid resuscitation and pain management of patients with fractured neck of femur', Levy N, Anaesthesia 2002; 57: 1148. PMID- 12648150 TI - A response to 'A reply', Cogliano M et al., Anaesthesia 2002; 57: 1235-6. PMID- 12648151 TI - A response to 'Not so free flowing', Kitsberg A, Agarwal S, Anaesthesia 2002; 57: 1143-4. PMID- 12648152 TI - A response to 'Convulsions following axillary brachial plexus blockade with levobupivacaine', Pirotta D, Sprigge J, Anaesthesia 2002; 57: 1187-9. PMID- 12648153 TI - A response to 'Learning to apply effective cricoid pressure using a part task trainer', Owen H et al., Anaesthesia 2002; 57: 1098-101. PMID- 12648154 TI - A response to 'Consent issues in obstetric anaesthesia', Roberts J, Anaesthesia 2002; 57: 1232-3. PMID- 12648155 TI - A response to 'Emergency physicians are not anaesthetists', Baigel GD, Anaesthesia 2002; 57: 1218-19. PMID- 12648158 TI - Regulation of cyclooxygenase-2 in renal medulla. AB - AIM: Renal medulla is a major site for production and action of prostaglandins (PGs). The renal medullary functions as well as structural integrity are in part dependent on PGs under certain physiological or pathophysiological conditions. The two COX isoforms, COX-1 (constitutive form) and COX-2 (inducible form) are both abundantly expressed in renal inner medulla at basal state, raising a question of which COX isoform may mediate the known functions of PGs in the region. As in many other cell types, COX-1 expression in renal medulla is unlikely subject to robust regulation. In contrast, COX-2 expression in renal medulla is markedly stimulated by chronic salt loading, dehydration and endotoxaemia in vivo. At cellular levels, the signalling pathways responsible for the COX-2 stimulation in renal medullary cells seem to involve both the mitogen activated protein kinases and NF-kappa B. It is likely that in response to various insults that are detrimental to renal medulla, the induction of PG synthesis may become more dependent on COX-2 than COX-1, and this phenomenon may be relevant to the cytoprotective response against the insults. PMID- 12648159 TI - Interactions between COX-2 and the renin-angiotensin system in the kidney. AB - AIM: In adult mammalian kidney, COX-2 expression is found in a restricted subpopulation of cells. The two sites of renal COX-2 localization detected in all species to date are the macula densa (MD) and associated cortical thick ascending limb cells (cTALH) and medullary interstitial cells. Physiological regulation of COX-2 in these cellular compartments suggests functional roles for eicosanoid products of the enzyme. In the MD region, COX-2 expression increases in high renin states [salt restriction, angiotensin converting enzyme (ACE) inhibition, renovascular hypertension], and selective COX-2 inhibitors significantly decrease plasma renin levels and renal renin activity and mRNA expression. An important role for COX-2-derived prostanoids in regulation of renin expression and secretion has also been determined by using mice with selective genetic deletion of either the COX-1 or COX-2 gene. There is evidence for negative regulation of MD/cTALH COX-2 by angiotensin II and by glucocorticoids and mineralocorticoids, suggesting that in the kidney, cortical COX-2 expression is regulated in part by alterations in activity of the renin-angiotensin system. PMID- 12648160 TI - Role of cyclooxygenase-2 in the control of renal haemodynamics and excretory function. AB - AIM: The available evidence supporting the importance of cyclooxygenase-2 (COX-2) in the regulation of renal haemodynamics and excretory function is summarized. Cyclooxygenase-2-derived metabolites play a very important role in regulating renal haemodynamics when sodium intake is low whereas it plays a minor role in the control of cortical blood flow when sodium intake is normal or elevated. The importance of COX-2 in the regulation of renal haemodynamics seems to be dependent on the endogenous production of other vasoactive products such as nitric oxide (NO) or noradrenaline. The activation of COX-2 in response to a decrease in NO may represent a mechanism aimed at defending the renal vasculature in the face of a decrease in NO levels. CONCLUSION: Contrary to the important role of COX-2 in the long-term regulation of renal haemodynamics, the metabolites derived from COX-2 seem to be only involved in the acute regulation of renal excretory function. PMID- 12648161 TI - Electroporation: theory and methods, perspectives for drug delivery, gene therapy and research. AB - Electroporation designates the use of short high-voltage pulses to overcome the barrier of the cell membrane. By applying an external electric field, which just surpasses the capacitance of the cell membrane, transient and reversible breakdown of the membrane can be induced. This transient, permeabilized state can be used to load cells with a variety of different molecules, either through simple diffusion in the case of small molecules, or through electrophoretically driven processes allowing passage through the destabilized membrane--as is the case for DNA transfer. Initially developed for gene transfer, electroporation is now in use for delivery of a large variety of molecules: From ions to drugs, dyes, tracers, antibodies, and oligonucleotides to RNA and DNA. Electroporation has proven useful both in vitro, in vivo and in patients, where drug delivery to malignant tumours has been performed. Whereas initial electroporation procedures caused considerable cell damage, developments over the past decades have led to sophistication of equipment and optimization of protocols. The electroporation procedures used in many laboratories could be optimized with limited effort. This review (i) outlines the theory of electroporation, (ii) discusses factors of importance for optimization of electroporation protocols for mammalian cells, (iii) addresses particular concerns when using electroporation in vivo, e.g. effects on blood flow and considerations regarding choice of electrodes, (iv) describes DNA electrotransfer with emphasis on use in the in vivo setting, and (v) sums up data on safety and efficacy of electroporation used to enhance delivery of chemotherapy to tumours in cancer patients. PMID- 12648162 TI - Increased contribution of alpha 1- vs. beta-adrenoceptor-mediated inotropic response in rats with congestive heart failure. AB - AIM: In failing myocardium the mechanical response to beta-adrenoceptor stimulation is attenuated. Alternative signalling systems might provide inotropic support when the beta-adrenoceptor system is dysfunctioning. Accordingly, the inotropic responses to alpha 1- and beta-adrenoceptor stimulation by the endogenous adrenoceptor agonist noradrenaline in non-failing and failing rat hearts were compared. METHODS: Chronic heart failure was induced in male Wistar rats by coronary artery ligation. Corresponding sham groups were prepared. After 6 weeks, papillary muscles from non-failing and failing hearts were isolated. Receptor binding studies were performed in the corresponding myocardium. The alpha 1-adrenoceptor-mediated inotropic response was not changed while the beta adrenoceptor-mediated response was substantially reduced in failing compared with non-failing myocardium. RESULTS: No change in potency for the agonists was observed at the alpha 1-adrenoceptors, while an increased potency for the agonists at the beta-adrenoceptors was found during heart failure. The lusitropic response to beta-adrenoceptor stimulation was intact during heart failure. No over all change in affinity or number of either adrenoceptor type was observed in receptor binding studies. The alpha 1-adrenoceptor-mediated inotropic response became dominating compared with the beta-adrenoceptor-mediated one in failing rat myocardium in contrast to the dominating role of the latter in non-failing myocardium. The attenuation of the beta-adrenoceptor-mediated inotropic response in rat failing myocardium was not because of a reduced number of receptors. CONCLUSION: Increasing contractility through stimulation of alpha 1-adrenoceptors in situ by the endogenous agonist may be an alternative way of inotropic support during heart failure and even more so during beta-adrenoceptor blockade. PMID- 12648163 TI - Effects of sex, gonadectomy, and oestrogen substitution on ischaemic preconditioning and ischaemia-reperfusion injury in mice. AB - AIM: Ischaemic preconditioning (IPC) has been demonstrated to protect heart function and viability, but has been predominantly studied in male animals. METHODS: We studied a possible influence of sex and oestrogen for protection in IPC. Infarct size and heart function after 40 min global ischaemia and 60 min reperfusion with or without preceding classic IPC was investigated in Langendorff perfused hearts. Hearts were harvested from 10-week-old male and female C57BL6 mice with or without gonadectomy 6 weeks earlier, or gonadectomy and substitution with 17 beta-oestradiol for 4 weeks (n = 104). RESULTS: Classic IPC reduced depression of left ventricular developed pressure (P < 0.01), attenuated the increase of end-diastolic pressure (P < 0.01), and reduced infarct size (P < 0.01) in hearts of untreated male mice, but failed to protect untreated females which had improved functional recovery and smaller infarctions than untreated males. After gonadectomy of female mice, developed pressure was reduced (P < 0.01) and infarct size increased (P < 0.01) compared with normal females, with no protection of preconditioning. The changes were not reversed by 17 beta oestradiol substitution. In hearts of gonadectomized males, the post-ischaemic increase of end-diastolic pressure was attenuated (P < 0.01), and enhanced after substitution with 17 beta-oestradiol (P < 0.01). The preconditioning effect disappeared after gonadectomy and gonadectomy with substitution in male mice. CONCLUSION: There is a sex difference in evoking preconditioning in male and female mice which is only partially dependent on sex hormones. PMID- 12648164 TI - Left ventricular volume changes during supine exercise in young endurance athletes. AB - AIM: The primary objective of the study was to measure the relative left ventricular volumes and the changes in left ventricular ejection fraction during supine position from rest to exercise in young endurance athletes. The secondary objective was to examine if there were gender differences regarding the volume reply and ejection fraction with exercise. METHOD: Sixty-five (35 female and 30 males) young healthy Swedish orienteers participated in the study. Left ventricular volume and ejection fraction changes between rest and submaximal supine bicycle exercise were measured with radionuclide ventriculography. RESULTS: The mean left ventricular end-diastolic volume increased by 13% (P < 0.001) but there was no change in end-systolic volume. Stroke volume was found to increase by 21% (P < 0.001). Left ventricular ejection fraction increased significantly (>0.04 units) in 54% of the athletes from rest to exercise; 5% of the athletes showed a decrease in ejection fraction. A negative correlation was found between ejection fraction at rest and the difference in ejection fraction from rest to exercise (r = -0.38, P = 0.002). There were no gender differences in the left ventricular volume changes or ejection fraction. CONCLUSION: During submaximal supine exercise, the adjustments in cardiac volumes in endurance athletes were small. There were no gender disparities concerning the left ventricular volume reply during exercise. PMID- 12648165 TI - Synthesis and degradation of type IV collagen in rat skeletal muscle during immobilization in shortened and lengthened positions. AB - AIM: Type IV collagen is a major protein in basement membranes surrounding and supporting skeletal muscle cells. In the present study, we tested the hypotheses that immobilization down-regulates synthesis and up-regulates degradation of type IV collagen in skeletal muscle. METHODS: mRNA level and concentration of type IV collagen as well as mRNA levels and activities of proteins involved in its degradation were analysed from soleus (SOL), gastrocnemius (GAS) and extensor digitorum longus muscles after immobilization in shortened and lengthened positions for 1, 3 and 7 days. RESULTS: Following immobilization, type IV collagen mRNA level was decreased in SOL and GAS suggesting down-regulated synthesis of this protein. The mRNA level and activity of matrix metalloproteinase-2 (proMMP-2) were increased in all muscles, while the activity of tissue inhibitor of metalloproteinase-2 was decreased in SOL and GAS. These findings reflect an increased capacity for degradation of type IV collagen. CONCLUSIONS: As a consequence of decreased synthesis/degradation ratio immobilization reduced the concentration of type IV collagen in all muscles. The regulation of type IV collagen through synthesis and/or degradation seems, however, to be muscle specific. Immobilization in lengthened position seems to delay and partly decrease the net degradation of type IV collagen. PMID- 12648166 TI - Comparison of force-velocity relationships of vastus lateralis muscle in isokinetic and in stretch-shortening cycle exercises. AB - AIM: This study investigated the force-velocity characteristics of the vastus lateralis (VL) muscle fascicle and muscle-tendon unit (MTU) in isolated lengthening and shortening actions, and during natural movement. METHODS: Four subjects performed maximal eccentric and concentric knee extensions (60, 120 and 180 degrees s-1). Unilateral counter movement jumps and drop jumps in the sledge apparatus served as natural movements. Vastus lateralis fascicle lengths were determined from ultrasonography. In vivo patellar tendon forces (PTF) were measured with an optic fibre technique. Patellar tendon force was derived to VL force according to the cross-sectional area of the muscle. Force in the direction of fascicle was calculated by dividing the VL force value by cosine of the fascicle angle. Force-velocity curves were constructed using angle specific values from isokinetic knee extensions (classical curve) and using instantaneous values from jumping exercises. RESULTS: In the fascicle level, we did not find an enhanced muscle force in the jumping performances as compared with the classical force-velocity curve. In the muscle-tendon level, the instantaneous force at high muscle-tendon shortening speeds exceeded that extrapolated according to Hill's equation. CONCLUSION: This difference between fascicle and muscle-tendon behaviour suggests that the neural input in fast stretch-shortening cycle exercises minimizes the length changes in muscle fascicle and enables storage and recoil of energy from elastic components that contributes to the enhanced mechanical output of the MTU during the push-off phase. PMID- 12648167 TI - Mathematical analysis of atelectasis formation in middle ears with sealed ventilation tubes. AB - AIM: A mathematical model was developed to identify time periods of atelectasis induction in middle ear (ME) ventilated via ventilating tubes (VT). Atelectatic ears are characterized by a total gas pressure lower than 760 mmHg. METHODS: Ventilating tubes were deliberately sealed and ME gas content changed in the presence of a preset blood gas pressure. Once sealed, CO2 rapidly diffuses out of the blood via lining tissues into the ME cleft. This results in initially a total ME pressure rise followed by a decrease in subatmospheric pressures. Time periods for atelectasis reformation was determined once ME pressure crossed the 760 mmHg value and continued to decline as the atelectasis reached higher grades. RESULTS: Time periods calculated by the model varied from 18 to 125 min in ME cavities ranging in volume from 0.5 to 3.5 mL, respectively. These results were calculated for conditions of venous blood in the lining mucosa blood and are consistent with prior clinical tests that measured an induced return to previous atelectasis state following the closure of the VT in 33 tested ears within 25-120 min (43 min on average). CONCLUSIONS: The model demonstrates that under the above conditions, diffusive gas transfer in relation to blood gas content is the leading mechanism to alterations in ME pressure and volume. It may be used as a tool to determine ME physiological cavity volume of ears with VT. PMID- 12648168 TI - Somato-sympathetic vasoconstriction to intranasal fluid administration with consecutive decrease in nasal nitric oxide. AB - AIM: Patients suffering from non-allergic chronic rhinosinusitis (NACRS) increasingly use intranasal saline sprays. They report better nasal comfort. METHODS: In order to better understand this phenomenon, we studied intranasal laser Doppler flowmetry (LDF) and nasal nitric oxide (NO) variations evoked by local administration of saline, histamine, N-acetylcysteine (NAC) and lidocaine at room temperature (22 degrees C). RESULTS: There was a significant (P < 0.05) 14 +/- 3.8% decrease in LDF signal after 30 s, which lasted for 60-90 s, for all the substances applied at 22 degrees C. This pharmaco-independent vasoconstriction was further studied in patients under general anaesthesia (GA), with saline at 37 degrees C and after intranasal adrenaline treatment. While GA did not influence the vasoconstriction, saline at 37 degrees C and adrenaline pre treatment abolished it. Nasal NO is influenced by vasoconstriction. Therefore we investigated, whether the observed vasoconstriction also changes nasal NO. A significant (P < 0.001) 8.03 +/- 0.59% decrease in nasal NO was recorded 60 s after administration of all the substances, and under GA after 22 degrees C saline application. This NO decrease was absent after intranasal adrenaline pre treatment. An additional experiment tested the effect of nose blowing on nasal NO concentration. We registered an NO decrease with a similar pattern than observed with the other substances. CONCLUSIONS: Intranasal fluid nebulization at 22 degrees C induces a sympathetic mediated, transient vasoconstrictor reflex response. This somato-sympathetic vasoconstriction induces a decrease in nasal NO. Both could be related to the subjective comfort experienced by NACRS patients using intranasal saline sprays. PMID- 12648169 TI - Occurrence of gestational diabetes mellitus and the value of different screening indicators for the oral glucose tolerance test. AB - BACKGROUND: The objective of the present study was to determine prevalence of gestational diabetes mellitus (GDM) in terms of impaired glucose tolerance (IGT) and diabetes mellitus (DM), and the value of traditional anamnestic risk factors for predicting outcome of the oral glucose tolerance test (OGTT). METHODS: A prospective population-based study in a defined geographic area in Sweden. All pregnant nondiabetic women (n = 4918) attending maternal health care from July 1994 to June 1996 were offered a 75g OGTT in gestational weeks 28-32. Traditional anamnestic risk factors, as well as results of the OGTT in terms of fasting-B glucose and 2h-B-glucose, were registered. RESULTS: 3616 (73.5%) women agreed to perform the OGTT. Sixty-one (1.7%) of those had GDM [47 (1.3%) had impaired glucose tolerance and 14 (0.4%) had diabetes mellitus]. 15.8% fulfilled traditional risk factor criteria. Traditional anamnestic risk factors as an indicator to perform an OGTT identified 29/61 GDM women and 9/14 women with DM. Among primiparas, 4/21 with gestational diabetes mellitus were detected. CONCLUSION: Using traditional risk factors as an indicator to perform an OGTT gives a low sensitivity to detect GDM and even DM especially among primiparas. PMID- 12648170 TI - Unexplained antepartum fetal death in Norway, 1985-97: diagnostic validation and some epidemiologic aspects. AB - OBJECTIVE: To validate the diagnosis of an unexplained antepartum fetal death in the Medical Birth Registry of Norway against data obtained from hospital records, alone and combined with autopsy data. To compare epidemiologic characteristics of an unexplained fetal death based on cases recorded by the three data sources. METHODS: Data on unexplained fetal deaths in the Registry were compared with clinical and autopsy data from 108 457 singletons with a gestational age >or= 28 weeks or a birthweight >or= 1000 g delivered in 1985-97 at Haukeland Hospital in Bergen and Aker Hospital in Oslo. RESULTS: Compared with clinical data, the positive and negative predictive values of a Registry diagnosis of an unexplained fetal death were 88% and 86%, respectively, while the sensitivity and specificity were 76% and 93%, respectively. Compared with clinical and autopsy data combined, the positive and negative predictive values of a Registry diagnosis of an unexplained fetal death were 77% and 89%, respectively, while the sensitivity and specificity were 78% and 88%, respectively. High agreement was observed in comparisons between the data sources of risks according to various independent variables. CONCLUSIONS: The validity of a diagnosis of an unexplained antepartum fetal death based on the Medical Birth Registry of Norway is sufficiently high to justify future large-scale epidemiologic studies based on this database. PMID- 12648171 TI - Evaluation of an Internet-based database on infectious disorders during pregnancy: INFPREG. AB - OBJECTIVE: To evaluate the use of an Internet-based information system on infectious disorders (INFPREG) in antenatal care in Sweden. METHODS: A postal questionnaire was sent to all antenatal clinics in Sweden in 2000 (n = 515) and 2002 (n = 503). The questionnaire consisted of sections covering use of computers, availability of Internet connections and the use of INFPREG in patient care. RESULTS: We received 404 completed questionnaires in 2000 and 501 in 2002. In 2000, 81% of the midwives had access to computers at their antenatal clinics, and this number had increased to 93% in 2002. Sixty-eight percent and 88% in 2000 and 2002, respectively, had computers with an Internet connection. Of the responding midwives, 74% in 2000 and 84% in 2002 had received information concerning INFPREG. In 2000, 29% of the midwives had visited INFPREG and this figure had increased to 58% in 2002. Of the midwives that had used INFPREG, 67% in 2000 and 81% in 2002 reported that the information obtained from the site was implemented in the patient care. Of the responders, 45% in 2000 and 43% in 2002 claimed that they needed more information on how to use INFPREG. CONCLUSIONS: A majority of midwives at antenatal clinics in Sweden have access to the Internet and are confident in using an Internet-based knowledge center on infectious disorders in pregnancy. The present study indicates a rapid acceptance among health care providers in antenatal care in Sweden of this new method for dissemination of information and guidelines. However, many midwives still want more information and knowledge on how to use an Internet-based information system. PMID- 12648172 TI - Microbial invasion and cytokine response in amniotic fluid in a Swedish population of women in preterm labor. AB - BACKGROUND: Previous studies indicate an association between intra-amniotic microbial invasion and/or inflammation and spontaneous preterm birth, but there is a limited amount of data available from Europe. The aim of this study was to investigate the occurrence of intra-amniotic microorganisms and cytokines (interleukin-6 and interleukin-8) in a Swedish population of women in preterm labor and their correlation with preterm birth. METHODS: Amniotic fluid was retrieved transabdominally from 61 patients in preterm labor before 34 weeks of gestation. Polymerase chain reaction analyses for Ureaplasma urealyticum and Mycoplasma hominis and culture for aerobic and anaerobic bacteria were performed. Interleukin-6 and interleukin-8 were analyzed with enzyme-linked immunosorbent assay. RESULTS: Microorganisms in amniotic fluid were detected in 10 patients (16%). Patients with detected bacteria in the amniotic fluid had significantly higher levels of interleukin-6 and interleukin-8. There was also an association between interleukin-6/-8, the amniocentesis-delivery interval (or= 1.5 ng/mL or interleukin-8 >or= 1.3 ng/mL was associated with an increased risk of delivery within 7 days (interleukin-6: relative risk 7.3; 95% confidence interval: 2.8-19; sensitivity 83%, specificity 87%; interleukin-8: relative risk 14, 95% confidence interval: 3.6-55, sensitivity 91%, specificity 87%). CONCLUSIONS: The occurrence of intra-amniotic microbial invasion and inflammation in this population of Swedish women in preterm labor was similar to data reported from populations with a higher incidence of preterm delivery. Amniotic interleukin-6 and interleukin-8 correlated with the presence of microorganisms and with preterm birth. PMID- 12648173 TI - Serial serum ferritin estimation in pregnant women at risk of preterm labor. AB - BACKGROUND: To estimate serial serum ferritin (SF) concentrations after 20 weeks of pregnancy in women at risk of having a preterm labor and to correlate them with the outcome of pregnancy. METHODS: Venous blood samples were drawn serially at 26, 30 and 34 weeks of gestation in one hundred pregnant women who were at risk of having a preterm delivery. Samples were analyzed for hemoglobin (Hb), leukocyte count, hemotocrit serum iron (SI), total iron-binding capacity (TIBC), SF and transferrin saturation. Seventy-six subjects delivered at term and 24 subjects delivered before 37 weeks. The various parameters were compared between the term and the preterm groups. RESULTS: Mean SF at different gestations in the term and the preterm group was not statistically different. Serial SF levels in subjects in the term group showed a declining trend with advancing gestation as seen in normal pregnancies. However, in the preterm group, a rising trend was observed. Further, SF levels of > 30 micro g/dl at 26 weeks and > 40 micro g/dl at 34 weeks were found to have a reasonable sensitivity and specificity for predicting preterm delivery. CONCLUSION: A SF concentration of > 40 micro g/dl and a rise in SF concentration with increasing gestation should alert the clinician regarding the possibility of preterm delivery. PMID- 12648174 TI - Dinoprostone versus misoprostol: a randomized study of nulliparous women undergoing induction of labor. AB - BACKGROUND: The objectives of the study were to compare the efficacy and safety of intravaginal misoprostol and intravaginal dinoprostone for induction of labor and to quantify the clinical response to suspicious cardiotocographic (CTG) readings. METHODS: One hundred and ninety-one patients were randomized to receive either 50 micro g misoprostol initially then a further identical dose 6 h later or 2 mg dinoprostone initially followed by 1 mg 6 h later, over a period of 24 h. If not in labor after 24 h, then both arms of the study would thereafter receive dinoprostone alone as per hospital protocol. RESULTS: The induction to delivery interval (1047 vs. 1355 min, p = 0.01), delivery within 12 h (35.4% vs. 18.9%, p = 0.02) and delivery within 24 h (83.3% vs. 63.3%, p = 0.01) were all shorter in the misoprostol arm. There were no differences in rates of oxytocin augmentation (p = 0.47), tachysystole (p = 0.32) and hyperstimulation syndrome (p = 0.82). There was an increase in the median number of times a doctor was called to advise on a suspicious CTG in the misoprostol group (1 vs. 2 occasions, p = 0.052), but there was no difference in neonatal outcome. CONCLUSIONS: Intravaginal misoprostol led to a shorter, more efficient labor, and although there was more anxiety related to the CTG, there was no increase in neonatal adverse effects. PMID- 12648175 TI - Randomized trial in multiparous patients: investigating a single vs. two-dose regimen of intravaginal misoprostol for induction of labor. AB - BACKGROUND: Multiparous patients have a higher risk of hyperstimulation and uterine rupture than nulliparous patients. The minimum possible dose of uterotonic drug should be used in induction of labor for multiparous patients to avoid excessive uterine activity, which could increase both maternal and fetal risks. METHODS: One hundred and four women were randomized to either a single dose of 50 micro g of intravaginal misoprostol in 24 h, or two consecutive doses of intravaginal 50 micro g misoprostol 6 h apart. RESULTS: The mean induction to delivery interval (789 min [95% CI: 637-941] vs. 576 min [95% CI: 484-667], p = 0.018) and delivery rate within 12 h (63% vs. 83%, p = 0.035) were higher in the two-dose group. The oxytocin augmentation rate (14% vs. 2%, p = 0.03) was higher in the single-dose group. There was a higher rate of clinician input related to suspicious cardiotocographic readings in the single-dose arm (p = 0.04). There was no statistical difference (p > 0.05) between the one- and two-dose regimens with respect to the rates of tachysystole (21% vs. 15%), hyperstimulation (3.9% vs. 0%), and meconium staining at delivery (9.8% vs. 13.2%). CONCLUSIONS: The two dose regimen was most efficient, but both regimens were well tolerated by the fetuses. PMID- 12648176 TI - Effects of epidural analgesia during labor on pelvic floor function after vaginal delivery. AB - BACKGROUND: Aims of this study were to determine the rate of symptoms related to perineal trauma (anal and stress urinary incontinence) and to assess pelvic floor muscle function in women who underwent epidural analgesia. METHODS: Comparative design comprising 70 matched pairs of primiparous mothers. Each woman was questioned about urogynecologic symptoms and examined by digital test, vaginal perineometry and uroflowmetric stop test score 3 months after vaginal delivery. Urogenital prolapse was defined in accordance with the Baden and Walker's 'Halfway System Classification'. Statistical analysis was performed using Fisher's exact test to compare the two groups and simple logistic regression models to estimate the odds ratios of every variable considered in respect of the control population. RESULTS: No significant difference was found in the incidence of stress urinary incontinence, anal incontinence and vaginal prolapse in the two study groups. No significant differences were found between the study groups with regard to the digital test, vaginal manometry and urine stream interruption test. CONCLUSIONS: Use of epidural analgesia is not associated with symptoms related to perineal trauma and pelvic floor muscle weakness. PMID- 12648177 TI - Shoulder dystocia and brachial plexus injury: a case-control study. AB - OBJECTIVE AND BACKGROUND: To evaluate risk factors for shoulder dystocia and brachial plexus injury using a case-control study at the departments of obstetrics and gynecology at the four largest hospitals in southern Sweden. All cases of shoulder dystocia between 1987 and 1993 inclusive were identified. For each case, two control infants with similar birthweight (+/- 100 g) and identical year of birth were randomly selected. METHODS: Original maternal records were reviewed and information regarding 10 potential risk factors was extracted. Odds ratios (ORs) were calculated using the Mantel-Haenszel method. Stratification was made for year of delivery, parity (0, I, II, III+), and maternal age (5-year class). RESULTS: In all, 107 infants with shoulder dystocia and 198 controls were included. The OR was greater than unity for all risk factors except gestational age. Three of the risk factors, induction of labor, epidural analgesia, and instrumental delivery, reached statistical significance. Thirty-four infants also suffered brachial plexus injury, giving a brachial plexus injury rate of 32% among the shoulder dystocia cases. We also made a separate analysis of the nine risk factors for brachial plexus injury following a shoulder dystocia, however none reached statistical significance. CONCLUSION: In this case-control study based on more than 100,000 deliveries at four large hospitals during a 7-year period, induction of labor, epidural analgesia, and instrumental delivery turned out to be significant risk factors for shoulder dystocia. For brachial plexus injury following shoulder dystocia, no significant risk factor was identified. PMID- 12648178 TI - Personality factors and emotional responses to pregnancy among IVF couples in early pregnancy: a comparative study. AB - BACKGROUND: Little is known as to whether negative experiences associated with infertility and IVF treatment persist after successful treatment. The aim of the study was to compare couples who have conceived after IVF and couples who have conceived naturally regarding personality factors and emotional responses to pregnancy. METHODS: Fifty-seven women pregnant after IVF and 55 male partners and 43 women who had conceived naturally and 39 male partners were recruited from university IVF clinics and antenatal clinics in Stockholm. The subjects were interviewed about their socio-demographic background. They completed scales of personality traits, anxiety, emotional responses to pregnancy, marital adjustment and reactions to recalled infertility while in pregnancy week 13 (range 11-17). RESULTS: The results showed that the IVF women had more muscular tension and were more anxious about loosing the pregnancy than the control women. The IVF women with high infertility distress were more anxious about loosing the pregnancy and less ambivalent than the women with lower distress. The IVF men had more somatic anxiety, indirect aggression, guilt, and were more detached and more anxious about loosing the pregnancy and less ambivalent than the control men. The IVF men with high infertility distress were more anxious about the baby not being normal than the men with lower infertility distress. CONCLUSIONS: The women and men who had conceived after IVF differed on a number of personality dimensions and emotional responses to the pregnancy from that of the women and the men who had conceived naturally. The results suggest that IVF couples may need additional emotional support in early pregnancy. PMID- 12648179 TI - Long CAG repeats in the AR gene are not associated with infertility in Finnish males. AB - BACKGROUND: The modulatory domain of the human androgen receptor gene contains a polymorphic CAG repeat coding for a polyglutamine tract. The length of the polyglutamine tract is inversely correlated with transcriptional activity of the androgen receptor. As androgens are crucial to spermatogenesis, decreased transcriptional activity of the androgen receptor associated with a long polyglutamine tract could lead to failure in spermatogenesis. Accordingly, long CAG repeats within the normal range have been suggested to be more common in infertile males than in the control population. METHODS: To test this hypothesis, we examined the CAG repeat number of 192 Finnish males with moderate or severe spermatogenic failure and 149 control males. RESULTS: Our results did not support the hypothesis, the controls harbored slightly longer CAG repeats than the infertile males. CONCLUSION: At least in the present study population from Finland, long CAG repeats in the androgen receptor gene do not play a significant role in spermatogenic failure. PMID- 12648180 TI - Subdermal progestin implant (Nestorone) in the treatment of endometriosis: clinical response to various doses. AB - OBJECTIVE: To evaluate the efficacy of three different doses of Nestorone progestin administered by implants for relief of pain, as well as bleeding patterns and symptoms in women with endometriosis. DESIGN: An open clinical study without a control group. PATIENTS: Twenty-one women in whom endometriosis was diagnosed and treated at laparoscopy or at laparotomy. INTERVENTIONS: The patients were randomly allocated to one of three groups receiving a daily dose of either 150 micro g, 200 micro g or 400 micro g Nestorone for two to four implants inserted subcutaneously for 7 months. After removal of the implants the follow-up period was 6 months. Six patients in the 150 micro g group, seven in the 200 micro g group and five in the 400 micro g group completed the study. MEASUREMENTS: Records of bleeding and registered side-effects and endometriosis related symptoms were kept by the patients. Ovarian status was evaluated by vaginal ultrasonography. Serum concentrations of Nestorone and of lipids were measured. RESULTS: Pelvic pain decreased significantly in response to the treatment in all groups but returned to pretreatment levels during the post treatment period. Bleeding and spotting was the most common side-effect followed by hypoestrogenic and psychic symptoms, with no significant difference among the groups. No significant changes in the serum levels of total, HDL or LDL cholesterol were found. One patient during the treatment period and four patients during the follow ups underwent surgical procedures. CONCLUSION: All three doses of Nestorone effectively alleviated endometriosis-related pain. While the Nestorone implant provides an interesting option for the treatment of endometriosis-related pain, its role needs to be assessed in comparative trials. PMID- 12648181 TI - Decreased serum leptin levels in women with uterine leiomyomas. AB - OBJECTIVE: To analyze the possible involvement of leptin in uterine leiomyomas. STUDY DESIGN: Serum leptin levels, determined by radioimmunoassay, were compared in myomatic (n = 50) and the normal (n = 50) women. RESULTS: A significant correlation was found between serum leptin levels and body mass index in both the myomatic women (r = 0.76, p < 0.001) and the normal women (r = 0.56, p < 0.001). Serum leptin levels in the myomatic women (9.3 +/- 0.6 ng/mL) were significantly lower (p < 0.001) than those in the normal women (13.6 +/- 1.2 ng/mL). In addition, the ratios of serum leptin levels/body mass index in the myomatic women (0.38 +/- 0.02) were significantly lower than those in the normal women (0.57 +/- 0.04) (p < 0.001). A significant correlation was found between the ratios of serum leptin levels/body mass index and body mass index (r = 0.59, p < 0.001) in the normal women, but not in the myomatic women (r = 0.27, p = 0.061). CONCLUSION: The lower plasma leptin levels observed in the women with myomas were independent of body mass index, and unlike the normal women there was no significant up-regulation of leptin production in response to increased adiposity. PMID- 12648182 TI - Are complications after an induced abortion associated with reproductive failures in a subsequent pregnancy? AB - BACKGROUND: We have previously shown that induced abortions result in a slightly increased risk of spontaneous abortion and preterm delivery in subsequent pregnancies. Danish records show that approximately 4% of women who undergo surgically induced abortions have complications related to the procedure. We examined whether it was women who had the short-term complications that carried an excess risk of spontaneous abortion and preterm delivery in the subsequent pregnancy. METHODS: Two cohorts of women who had had an induced abortion and a subsequent pregnancy were followed. The cohort with no reported complications comprised 12,972 women, and the cohort with complications 605 women. RESULTS: The cohort with complications did not have more spontaneous abortions or preterm births. They did, however, have a higher risk of stillbirth, mainly seen in women whose induced abortion had been complicated by an infection. CONCLUSION: Most of the women who had had an induced abortion with reported complications did, however, carry a subsequent pregnancy to term with no more complications than other women of similar parity and age. PMID- 12648183 TI - Cigarette, alcohol, and caffeine consumption: risk factors for spontaneous abortion. AB - OBJECTIVE: To study the association between cigarette, alcohol, and caffeine consumption and the occurrence of spontaneous abortion. METHODS: The study population consisted of 330 women with spontaneous abortion and 1168 pregnant women receiving antenatal care. A case-control design was utilized; cases were defined as women with a spontaneous abortion in gestational week 6-16 and controls as women with a live fetus in gestational week 6-16. The variables studied comprise age, parity, occupational situation, cigarette, alcohol, and caffeine consumption. The association between cigarette, alcohol, and caffeine consumption was studied using logistic regression analyzes while controlling for confounding variables. In addition stratified analyzes of the association between caffeine consumption and spontaneous abortion on the basis of cigarette and alcohol consumption were performed. RESULTS: Women who had given birth twice or more previously had increased odds ratio (OR), 1.78 (1.27-2.49), whereas women who were students had decreased OR, 0.55 (0.34-0.91) for having spontaneous abortions. Regarding lifestyle factors, the adjusted ORs among women who consumed 5 units or more alcohol per week or 375 mg or more caffeine per day were 4.84 (2.87-8.16) and 2.21 (1.53-3.18), respectively. Women who smoked 10-19 cigarettes and 20 or more cigarettes per day did not have significantly increased ORs for having spontaneous abortions, after adjusting for other risk factors. CONCLUSION: Consumption of 5 or more units alcohol per week and 375 mg or more caffeine per day during pregnancy may increase the risk of spontaneous abortion. PMID- 12648184 TI - Endoscopic treatment of interstitial pregnancy. PMID- 12648185 TI - High risk of cesarean section among ethnic Filipinos: an effect of the paternal contribution to birthweight? PMID- 12648186 TI - Coronary artery dissection during pregnancy. PMID- 12648187 TI - Ovarian Sertoli-Leydig cell tumor of intermediate differentiation with immature skeletal muscle heterologous elements. PMID- 12648188 TI - Serum-hCG: still a problematic marker. PMID- 12648189 TI - Gender differences in drug effects: implications for anesthesiologists. AB - BACKGROUND: The gender aspect in pharmacokinetics and pharmacodynamics of anesthetics has attracted little attention. Knowledge of previous work is required to decide if gender-based differences in clinical practice is justified, and to determine the need for research. METHODS: Basis for this paper was obtained by Medline searches using the key words 'human' and 'gender' or 'sex,' combined with individual drug names. The reference lists of these papers were further checked for other relevant studies. RESULTS: Females have 20-30% greater sensitivity to the muscle relaxant effects of vecuronium, pancuronium and rocuronium. When rapid onset of or short duration of action is very important, gender-modified dosing may be considered. Males are more sensitive than females to propofol. It may therefore be necessary to decrease the propofol dose by 30 40% in males compared with females in order to achieve similar recovery times. Females are more sensitive than males to opioid receptor agonists, as shown for morphine as well as for a number of kappa (OP2) receptor agonists. On this basis, males will be expected to require 30-40% higher doses of opioid analgesics than females to achieve similar pain relief. On the other hand, females may experience respiratory depression and other adverse effects more easily if they are given the same doses as males. CONCLUSION: These examples illustrate that gender should be taken into account as a factor that may be predictive for the dosage of several anesthetic drugs. Moreover, there is an obvious need for more research in this area in order to further optimize drug treatment in anesthesia. PMID- 12648191 TI - Catecholamine release during laparoscopic fundoplication with high and low doses of remifentanil. AB - BACKGROUND: Reports on stress responses to laparoscopic surgery have been conflicting. Depth of anesthesia may influence the neuro-hormonal release, including catecholamines. Opioids depress general sympathetic activation in a dose-dependent manner. We investigated the hypothesis that remifentanil would depress the catecholamine response to pneumoperitoneum and laparoscopic surgery differently with a high dose (HD) compared with a low dose (LD). METHODS: In a randomized, prospective study we investigated 18 ASA I-II patients undergoing laparoscopic fundoplication with an intra-abdominal pressure of 12 mmHg. The patients were randomized to receive either a LD (0.13 microg kg-1x min-1) or HD (0.39 microg kg-1 x min-1) of remifentanil with a target-controlled infusion (TCI) technique. Bispectral index of EEG (BIS) was maintained at 40-55 by propofol delivered by a TCI system. Arterial catecholamines were analyzed at different times during the procedure. RESULTS: Norepinephrine increased equally in both groups during pneumoperitoneum and surgical intervention. Epinephrine stayed low in the HD-group, while increasing during surgery in the LD-group. CONCLUSION: High dose of remifentanil depressed the epinephrine response to pneumoperitoneum and surgery, indicating no general activation of the sympathetic nervous system. Neither a LD nor HD of remifentanil depressed the norepinephrine response during pneumoperitoneum. This suggests a centrally independent release of norepinephrine. PMID- 12648192 TI - Retrograde intubation with a Mini-Trach II kit. AB - BACKGROUND: Retrograde intubation has been accepted internationally as a viable alternative for managing the difficult airway. Various techniques have been described to perform this procedure, however, difficulties have arisen on account of problems with suboptimal materials. We therefore describe a retrograde intubation technique using the knife and stiff plastic introducer from a Mini Trach II set from Portex Ltd (Kent, UK). METHODS: The cricothyroid membrane was identified and using the knife from the mini-trach set, incised longitudinally. The plastic introducer was inserted through the incision and maneuvered out through the mouth providing a guide over which the endotracheal tube was threaded. The technique was evaluated on 20 cadavers and thereafter used in four patients. RESULTS: Mean intubation time in the 20 cadavers was 6.7 s (range 3-10) from incision to removal of the guide. Also, the technique was used successfully in four patients in whom anterograde attempts failed. In one of these patients the retrograde intubation was life saving. CONCLUSION: Retrograde intubation with a stiff curved plastic introducer was rapid and easy in cadavers and in four patients. In emergency situations where conventional intubation fails it may be life saving. PMID- 12648190 TI - Does anaesthesia cause postoperative cognitive dysfunction? A randomised study of regional versus general anaesthesia in 438 elderly patients. AB - BACKGROUND: Postoperative cognitive dysfunction (POCD) is a common complication after cardiac and major non-cardiac surgery with general anaesthesia in the elderly. We hypothesized that the incidence of POCD would be less with regional anaesthesia rather than general. METHODS: We included patients aged over 60 years undergoing major non-cardiac surgery. After giving written informed consent, patients were randomly allocated to general or regional anaesthesia. Cognitive function was assessed using four neuropsychological tests undertaken preoperatively and at 7 days and 3 months postoperatively. POCD was defined as a combined Z score >1.96 or a Z score >1.96 in two or more test parameters. RESULTS: At 7 days, POCD was found in 37/188 patients (19.7%, [14.3-26.1%]) after general anaesthesia and in 22/176 (12.5%, [8.0-18.3%]) after regional anaesthesia, P = 0.06. After 3 months, POCD was present in 25/175 patients (14.3%, [9.5-20.4%]) after general anaesthesia vs. 23/165 (13.9%, [9.0-20.2%]) after regional anaesthesia, P = 0.93. The incidence of POCD after 1 week was significantly greater after general anaesthesia when we excluded patients who did not receive the allocated anaesthetic: 33/156 (21.2%[15.0-28.4%]) vs. 20/158 (12.7%[7.9-18.9%]) (P = 0.04). Mortality was significantly greater after general anaesthesia (4/217 vs. 0/211 (P < 0.05)). CONCLUSION: No significant difference was found in the incidence of cognitive dysfunction 3 months after either general or regional anaesthesia in elderly patients. Thus, there seems to be no causative relationship between general anaesthesia and long-term POCD. Regional anaesthesia may decrease mortality and the incidence of POCD early after surgery. PMID- 12648193 TI - Analgesia following music and therapeutic suggestions in the PACU in ambulatory surgery; a randomized controlled trial. AB - BACKGROUND: This study was designed to determine whether music (M), or music in combination with therapeutic suggestions (M/TS) could improve the postoperative recovery in the immediate postoperative in daycare surgery. METHODS: One-hundred and eighty-two unpremedicated patients who underwent varicose vein or open inguinal hernia repair surgery under general anaesthesia were randomly assigned to (a). listening to music (b). music in combination with therapeutic suggestions or (c). blank tape in the immediate postoperative period. The surgical technique, anaesthesia and postoperative analgesia were standardized. Analgesia, the total requirement of morphine, nausea, fatigue, well-being, anxiety, headache, urinary problems, heart rate and oxygen saturation were studied as outcome variables. RESULTS: Pain intensity (VAS) was significantly lower (P = 0.002) in the M (2.1), and the M/TS (1.9) group compared with the control group (2.9) and a higher oxygen saturation in M (99.2%) and M/TS (99.2%) group compared with the control (98.0%), P < 0.001, were found. No differences were noted in the other outcome variables. CONCLUSION: This controlled study has demonstrated that music with or without therapeutic suggestions in the early postoperative period has a beneficial effect on patients' experience of analgesia. Although statistically significant, the improvement in analgesia is modest in this group of patients with low overall pain levels. PMID- 12648194 TI - Inhibitory effect of propofol on ketamine-induced c-Fos expression in the rat posterior cingulate and retrosplenial cortices is mediated by GABAA receptor activation. AB - BACKGROUND: Non-competitive N-methyl-D-aspartate (NMDA) receptor antagonists, including ketamine, have psychotomimetic activities and cause neuronal damage in the posterior cingulate and retrosplenial cortices (PC/RS), which are suggested to be the brain regions responsible for their psychotomimetic activities. We previously demonstrated that ketamine induced marked c-Fos (c-fos protein) expression in the rat PC/RS, which was inhibited by propofol, and the expression was closely related to ketamine-induced abnormal behavior. In the present study, we investigated whether the inhibition by propofol was mediated by GABAA receptor receptor activation. METHODS: Using Wistar rats, propofol alone, propofol with bicuculline or propofol with flumazenil was injected intravenously and then continuously infused. Fifteen minutes later, 100 mg kg-1 of ketamine or normal saline was injected intraperitoneally. Two hours after the ketamine or saline injection, the brain was extracted and brain sections were prepared, and c-Fos expression was detected using immunohistochemical methods. RESULTS: Ketamine induced marked c-Fos expression in the PC/RS (171 +/- 9/0.4 mm2), which was significantly inhibited by propofol (5 +/- 5/0.4 mm2). The inhibition by propofol was disinhibited dose-dependently by both bicuculline (0.5 and 1.0 mg kg-1 bicuculline groups: 46 +/- 15 and 143 +/- 16, respectively) and flumazenil (0.1 and 1.0 mg kg-1 flumazenil groups: 79 +/- 6 and 130 +/- 15, respectively). CONCLUSION: These results demonstrate that the inhibitory effect of propofol on ketamine-induced c-Fos expression in the PC/RS is mediated by GABAA receptor activation, and suggests that ketamine-induced psychoneuronal adverse effects may be suppressed by propofol via the activation of GABAA receptors. PMID- 12648196 TI - Multiple organ failure and outcome of critically ill patients with haematological malignancy. AB - BACKGROUND: The number of failing organs systems in ICU patients with haematological malignancy is associated with outcome. The objective of this study was to assess short and long-term survival in these patients with special reference to multiple organ failure reflected by the SOFA (Sequential Organ Failure Assessment) score. METHODS: Retrospective chart review of haematological patients admitted to the 10-bed intensive care unit (ICU) of a tertiary level academic teaching hospital from 1994 to 1998. Of 31 admitted patients with the diagnosis of haematological malignancy, the charts of 30 were available for analysis. RESULTS: Univariate logistic regression analysis of factors previously shown to influence survival revealed that only admission SOFA score and untreated status of haematological disease were significantly associated with survival (P < 0.05). ICU, 3-month and one-year survival rates were 57% (17/30), 23% (7/30) and 20% (6/30), respectively. If maximal SOFA score during the ICU stay was included in a multivariate model comprising treatment status and effect, admission day SOFA and APACHE II scores, mechanical ventilation, renal replacement therapy and neutropenia, the maximal SOFA score became the only independent variable. All patients with an admission SOFA score exceeding 11 died in hospital. Leave-one out method revealed that admission SOFA scores and the status of haematological disease (untreated or not) correctly classified 83% (25 of 30) of patients to survivors or non-survivors. CONCLUSIONS: Multiple organ failure assessed as SOFA score on admission and status of disease were associated with outcome in critically ill patients with haematological malignancy. PMID- 12648195 TI - A new short-acting non-depolarizing muscle relaxant (SZ1677) without cardiovascular side-effects. AB - BACKGROUND: In order to facilitate rapid tracheal intubation, the development of a rapid onset, short duration, non-depolarizing muscle relaxant without cardiovascular side-effects would be a significant accomplishment in the field of anesthesiology. The aim of the present study was to test the action of a new non depolarizing muscle relaxant (SZ1677) on neuromuscular transmission, muscarinic (M2, M3) receptors and cardiovascular reactions and to compare it with clinically used muscle relaxants. METHODS: Neuromuscular transmission was studied by recording muscle contractions elicited by indirect electrical stimulation, using (i). in vitro isolated phrenic nerve-hemidiaphragm preparation of mice, rats and guinea pigs and (ii). in vivo sciatic nerve-anterior tibial muscle preparation of anesthetized rats, guinea pigs and cats. Cardiovascular effects of muscle relaxants were evaluated on the grounds of their effects on changes of blood pressure and heart rate induced by electrical stimulation of the right vagal nerve in anesthetized cats. To study postsynaptic antimuscarinic affinity of muscle relaxants on M3 receptors, oxotremorine-induced contractions of longitudinal muscle strip of guinea pig ileum were registered in their presence and absence. RESULTS: One of more than 120 newly synthesized non-depolarizing muscle relaxants compounds, 1-3[alpha-hydroxy-17beta-acetyloxy-2beta-(1,4-dioxa-8 azaspiro[4,5]dec-8-yl)-5alpha-androstane-16beta-il] -1-(2-propenyl)pyrrolidinium bromide (SZ1677), excelled with its advantageous pharmacological properties: relatively short duration of action, no accumulation and lack of unwanted side effects. Pharmacodynamic studies show that SZ1677 is a non-depolarizing neuromuscular blocking agent with a relatively short duration and rapid onset of action in a variety of laboratory animal species. It is without cumulative effect, does not reduce blood pressure, and fails to produce tachycardia. Significant cardiac vagal blocking effects were not observed even at concentrations or dosages of 8 times the ED90. This compound, unlike many other muscle relaxants, does not have atropine-like effects on human atrial tissue; it does not increase the release of NA from sympathetic innervation in the heart. In all practical ways, at least from the vantage point of the preclinical study, SZ1677 compares favorably with all presently available short-acting muscle relaxants, including rapacuronium. CONCLUSION: In experiments, SZ1677 proved to be a short-acting neuromuscular blocking compound having a large safety margin between the doses required to produce neuromuscular block and those likely to lead to cardiovascular side-effects. PMID- 12648198 TI - Nitric oxide increases dramatically in air exhaled from lung regions with occluded vessels. AB - BACKGROUND: We observed dramatic changes in exhaled nitric oxide concentration (DeltaNOE) during wedge measurements, and hypothesised that occlusion and redistribution of pulmonary blood flow affects NOE. METHODS: We inflated the balloon of the pulmonary artery catheter and measured NOE and central hemodynamics in closed chest anesthetised pigs (n = 11) ventilated with hyperoxic gas (fraction of inspired oxygen [FIO2] = 0.5), before and during lung injury, and in open chest anesthetised pigs (n = 17) before and during left lower lobar (LLL) hypoxia (FIO2 0.05), and during hyperoxic (FIO2 0.8) ventilation of the other lung regions (HL). RESULTS: In the closed chest pigs NOE increased from 2.0 (0.9) to 3.4 (2.0) p.p.b. (P < 0.001) during wedge, and returned to 2.0 (1.0) p.p.b. when the balloon was deflated. The increase in mean pulmonary artery pressure (MPaP) during wedge was small and insignificant (P > 0.07). When the balloon was inflated in the right pulmonary artery in the open chest pigs, the perfusion of the HL decreased from 2.57 (0.58) to 2.34 (0.55) l min(-1) (P < 0.001), and NOEHL increased from 2.5 (0.9) to 6.2 (3.2) p.p.b. (P < 0.001). The perfusion of the LLL increased from 0.33 (0.26) to 0.54 (0.34) l min(-1) (P < 0.001), and NOELLL decreased from 1.7 (0.6) to 1.5 (0.5) p.p.b. (P < 0.001). Neither lung injury nor LLL hypoxia had any influence on DeltaNOE (P > 0.07) during wedge. The correlation coefficient (R2) was 0.66 between changes in regional blood flow and DeltaNOE, and 0.37 between changes in MPaP and DeltaNOE. CONCLUSIONS: Nitric oxide concentration increases dramatically from lung regions with occluded vessels, whereas changes in MPaP have minor effects on NOE. This is an important fact to consider when comparing NOE within or between studies, and indicates a possible marker of diseases with occluded lung vessels. PMID- 12648197 TI - The effect of nitrous oxide on cerebral blood flow velocity in children anesthetized with propofol. AB - BACKGROUND: Propofol for maintenance of anesthesia by continuous infusion is gaining popularity for use in pediatric patients. Nitrous oxide (N2O) has been shown to increase cerebral blood flow velocity (CBFV) in both children and adults. To determine the effects of N2O on middle cerebral artery blood flow velocity (Vmca) during propofol anesthesia in children, Vmca was measured with and without N2O using transcranial Doppler (TCD) sonography. METHODS: Thirty ASA I or II children aged 18 months to 6 years undergoing elective urological surgery were enrolled. Anesthesia comprised propofol aimed at producing an estimated steady-state serum concentration of 3 micro g.ml-1 and a caudal epidural block. A transcranial Doppler probe was used to measure middle cerebral artery blood flow velocity. Each patient was randomized to receive a sequence of either Air/N2O/Air or N2O/Air/N2O in 35% oxygen. Fifteen min after each change in the N2O concentration, three measurements of cerebral blood flow velocity, blood pressure and heart rate were recorded. Ventilatory parameters and EtCO2 were kept constant throughout the study period. RESULTS: CBFV increased by 12.4% when air was replaced by N2O, and returned to baseline when N2O was subsequently removed. There was a 14% decrease in CBFV when N2O was replaced with air, which increased to baseline when air was subsequently replaced with N2O. Mean heart rate and blood pressure remained constant throughout the study period. CONCLUSION: The effects of nitrous oxide on CBFV are preserved in children during propofol anesthesia. PMID- 12648199 TI - Anti-inflammatory cytokine response and the development of multiple organ failure in severe sepsis. AB - BACKGROUND: The production of proinflammatory cytokines activates the systemic inflammatory response in sepsis. Patients also develop a compensatory anti inflammatory reaction, which may have an important down-regulatory effect on the overactive inflammation. However, the role of this anti-inflammatory response in sepsis is not completely clarified. In this prospective study, we investigated the relationship between the pro- and anti-inflammatory cytokine profiles in severe sepsis and their role in the development of multiple organ failure (MOF). METHODS: Thirty-eight patients meeting the criteria for severe sepsis were studied. MOF was defined as a maximum SOFA score of 10 or higher. Serial measurements of the proinflammatory IL-6 and IL-1beta and the anti-inflammatory IL-10 and IL-1ra were used. The cytokine samples were taken at the onset of sepsis and on the third and fifth day during the ICU period. RESULTS: The initial IL-10 and IL-1ra responses were identical in patients with or without MOF. The anti-inflammatory cytokine levels remained elevated in the MOF patients, whereas in patients without MOF the levels declined. The IL-6/IL-10 ratio was significantly higher in the MOF patients on days 1 and 3 compared with patients without MOF. CONCLUSIONS: We could not demonstrate overproduction of anti inflammatory IL-10 in MOF patients. On the contrary, the high IL-6/IL-10 ratio indicates that IL-10 deficiency may contribute to the development of MOF in severe sepsis. PMID- 12648200 TI - Adjusting positive end-expiratory pressure and tidal volume in acute respiratory distress syndrome according to the pressure-volume curve. AB - BACKGROUND: Management of acute respiratory distress syndrome (ARDS) patients implies the selection of the adequate ventilatory parameters, essentially PEEP and tidal volume (Vt), to prevent ventilator-induced lung injury. These parameters should be reset as the lung injury evolves. Among the different methods proposed for the adjustment of the ventilator, the measurement of the P-V curve has emerged as a useful, although debated, tool. Our aim has been to study the relationship between the different inflection points of the P-V curve in ARDS patients, and to assess the changes in the empiric PEEP and Vt (PEEP(emp), V(temp) following its use. METHODS: P-V curves were measured in 27 patients (lung injury score [LIS] >or= 2, 69 measurements) by means of the low-flow continuous inflation method. RESULTS: A lower inflection point (LIP) was found in all patients and, although it correlated with the PEEP(emp), there was only a fair concordance, so the PEEP was modified in 80% of the cases. The expiratory inflection point (EIP) was significantly lower than the LIP (6.3 +/- 1.7 vs. 8.1 +/- 3.2, P = 0.008). An upper inflection point was observed in 16 measurements (23%) and the Vt was reset in 20% of the cases. Both PEEP and Vt were readjusted on 10 occasions (14%). Only the EIP was significantly higher on the first 3 days of mechanical ventilation. The LIS was correlated with all the inflection points. There were no differences for any parameter independent of the cause of the ARDS (pulmonary/extrapulmonary). CONCLUSIONS: The quasi-static measurement of the P-V curve is a simple method, easy to interpret, for objective adjustment of the ventilatory parameters in ARDS patients as the lung injury evolves. The implementation of this strategy may vary the empiric clinical practice. The role of the EIP for the evaluation of the severity of lung injury deserves further investigation. PMID- 12648201 TI - Chronic intraperitoneal endotoxin treatment in rats induces resistance to d tubocurarine, but does not produce up-regulation of acetylcholine receptors. AB - BACKGROUND: Chronic systemic inflammation resulting from intraperitoneal Eschevichia coli endotoxin administration or Corynebacterium injections induces tolerance to non-depolarizing neuromuscular blockers in rodents. Although this has been explained as up-regulation of muscle acetylcholine receptors (AChR), the numbers of involved receptors have not been documented. The aim of this study was to determine the effects of chronic endotoxin administration on rat muscle AChR. METHODS: One day after one, seven, or 14 daily intraperitoneal doses of lipopolysaccharide endotoxin (0 or 0.5 mg kg(-1)), we studied in vivo dose response relationships for d-tubocurarine (d-Tc) and AChR binding using [125I]alpha-bungarotoxin as a ligand. RESULTS: One day after seven and 14 daily intraperitoneal doses of endotoxin, the effective dose of d-Tc required to suppress the twitch response to 50% of the control (ED50) was significantly increased compared with that of time-matched control rats (146.5 +/- 38.2 vs. 76.1 +/- 9.0 microg kg(-1) for seven doses; 116.4 +/- 51.3 vs. 74.4 +/- 9.6 micro g kg-1 for 14 doses, P < 0.05). However, this was not associated with an increase in the number of AChR in the anterior tibial muscle or diaphragm. CONCLUSIONS: Mechanisms other than AChR up-regulation might be responsible for the increased d Tc requirement during chronic intraperitoneal endotoxin administration. PMID- 12648202 TI - Intrathecal hyperbaric bupivacaine 3 mg + fentanyl 10 microg for outpatient knee arthroscopy with tourniquet. AB - BACKGROUND: Combination of local anesthetic and opioid enables the use of less spinal anesthetic and increases the success of anesthesia. Intrathecal opioid does not prolong motor recovery and thus should not delay discharge home. We hypothesized that 3 mg of hyperbaric bupivacaine with 10 microg of fentanyl permits fast-tracking or shorter stay in post anesthesia care unit (PACU), and earlier discharge home, compared with 4 mg of hyperbaric bupivacaine. METHODS: In this double-blind study, 100 outpatients undergoing knee arthroscopy received randomly either 4 mg of bupivacaine (B4) or 3 mg of bupivacaine + 10 microg fentanyl (B3F) intrathecally. The volume of 0.8 ml was injected at the L2/3 interspace over a 2-min period. A lateral decubitus position was maintained for 10 min. The sensory block was recorded by using thermal stimuli, and motor block was assessed according to a modified Bromage scale. Fast-tracking criteria were complete recovery of motor block, sensory block Th12 or lower and stable vital signs. RESULTS: One block (1%) failed. Motor recovery was faster in the B3F group: 60% of the patients recovered in 80 min or less compared with 28% in group B4 (P = 0.002). The PACU-time was shorter: 36 (10-103) vs. 55 (10-140) min, respectively (P = 0.005). Seventeen (B3F) vs. nine patients (B4) could bypass PACU (NS). Time to discharge home was similar in both groups. In the B3F group, 75% of the patients developed pruritus. CONCLUSION: Both solutions produced reliable spinal anesthesia for outpatient knee arthroscopy. The PACU-time was shorter in the bupivacaine-fentanyl group, but both groups reached home-readiness equally. PMID- 12648203 TI - Effects of intrathecally administered dexmedetomidine, MPV-2426 and tizanidine on EMG in rats. AB - BACKGROUND: When administered intrathecally, alpha-2 adrenergic agonists produce spinally mediated antinociception, but also rapidly redistribute to supraspinal sites. This investigation the compared EMG effects of intrathecally administered dexmedetomidine, MPV-2426 (fadolmidine), and tizanidine in Sprague-Dawley rats, which has not been previously described. METHODS: We studied electromyographic (EMG) responses of the head and gastrocnemius muscles, antinociception using the tail-flick test, and sedation by using observer assessment. Saline, dexmedetomidine (0.5 microg, 2.5 microg and 12.5 microg), MPV-2426 (2 microg, 10 microg and 50 microg) and tizanidine (2 microg, 10 microg and 50 microg) were administered intrathecally. RESULTS: Tizanidine 50 microg, MPV-2426 10 microg and 50 microg, and dexmedetomidine 2.5 microg and 12.5 microg, decreased EMG activity (P < 0.005). Dexmedetomidine 12.5 microg, MPV-2426 50 microg, and tizanidine 10 microg and 50 microg increased tail-flick latencies (P < 0.01). Dexmedetomidine alone significantly increased the magnitude of observer-assessed sedation (P < 0.0001). CONCLUSION: We conclude that in rats, intrathecally administered dexmedetomidine, MPV-2426 and tizanidine have dose-dependent effects on EMG. At antinociceptive doses, the EMG effects of these three alpha-2 adrenergic agonists differ (dexmedetomidine > MPV-2426 > tizanidine). PMID- 12648204 TI - Lidocaine versus ropivacaine for continuous interscalene brachial plexus block after open shoulder surgery. AB - BACKGROUND: This study compared the postoperative infusion of 1% lidocaine and 0.2% ropivacaine for continuous interscalene analgesia in patients undergoing open shoulder surgery. METHODS: Forty patients undergoing open shoulder surgery received an interscalene brachial plexus block with 30 ml of either 1.5% lidocaine (n = 20) or 0.5% ropivacaine (n = 20), followed by a continuous patient controlled interscalene analgesia with 1% lidocaine or 0.2% ropivacaine, respectively. A blinded observer recorded the quality of analgesia and recovery of motor function during the first 24 h of infusion. RESULTS: Onset of the block occurred after 7.5 (5-40) min with lidocaine and 30 (10-60) min with ropivacaine (P = 0.0005). Postoperative pain intensity was higher with lidocaine than ropivacaine for the first 8 h of infusion. The ratio between boluses given and demanded from the pump was 0.5 (0.13-0.7) with lidocaine and 0.7 (0.4-1.0) with ropivacaine (P = 0.005). Rescue IV tramadol was required during the first 24 h of infusion by 16 patients of the lidocaine group (84%) and eight patients of the ropivacaine group (46%) (P = 0.05). At the 16 h and 24 h observation times a larger proportion of patients receiving ropivacaine had complete regression of motor block (70% and 95%) than patients receiving lidocaine (50% and 55%) (P = 0.05 and P = 0.013, respectively). CONCLUSIONS: Although 1% lidocaine can be effectively used for postoperative patient-controlled interscalene analgesia, 0.2% ropivacaine provides better pain relief and motor function. PMID- 12648205 TI - Nitrous oxide anesthesia and intravitreal gastamponade. AB - The right eye of a 66-year-old man was operated with vitrectomy and peeling of an epiretinal membrane. Perioperatively, the eye was filled with 20% SF6 gas to tamponade retinal breaks. Five days later the patient underwent prostatectomy under general anesthesia using nitrous oxide. Postoperatively the eye had no light perception as a result of ischemic retinopathy. The movement of nitrous oxide into gas-containing spaces in the body has been known for a long time. The use of nitrous oxide in patients with intravitreal gas will elevate the intraocular pressure with risk for closure of the central retinal artery. The present case report highlights the problems that can occur when preoperative assessment is carried out a long time before surgery. PMID- 12648206 TI - Cardiopulmonary resuscitation after near drowning and hypothermia: restoration of spontaneous circulation after vasopressin. AB - Recent animal data have challenged the common clinical practice to avoid vasopressor drugs during hypothermic cardiopulmonary resuscitation (CPR) when core temperature is below 30 degrees C. In this report, we describe the case of a 19-year-old-female patient with prolonged, hypothermic, out-of-hospital cardiopulmonary arrest after near drowning (core temperature, 27 degrees C) in whom cardiocirculatory arrest persisted despite 2 mg of intravenous epinephrine; but, immediate return of spontaneous circulation occurred after a single dose (40 IU) of intravenous vasopressin. The patient was subsequently admitted to a hospital with stable haemodynamics, and was successfully rewarmed with convective rewarming, but died of multiorgan failure 15 h later. To the best of our knowledge, this is the first report about the use of vasopressin during hypothermic CPR in humans. This case report adds to the growing evidence that vasopressors may be useful to restore spontaneous circulation in hypothermic cardiac arrest patients prior to rewarming, thus avoiding prolonged mechanical CPR efforts, or usage of extracorporeal circulation. It may also support previous experience that the combination of both epinephrine and vasopressin may be necessary to achieve the vasopressor response needed for restoration of spontaneous circulation, especially after asphyxial cardiac arrest or during prolonged CPR efforts. PMID- 12648207 TI - Uvulopalatopharyngoplasty for sleep apnea in mentally retarded obese 14-year-old: an anaesthetic challenge. AB - Anaesthetic management of patients with obstructive sleep apnea for upper airway surgery has always been a challenging task. We report our anaesthetic approach for a young, mentally retarded obese patient with documented obstructive sleep apnea undergoing uvulopalatopharyngoplasty. The therapeutic intervention before, during and after operation is discussed. PMID- 12648208 TI - Anesthetic management of a patient with Weill-Marchesani syndrome. AB - Weill-Marchesani syndrome is characterized by short stature, brachydactylyl, myopia, microspherophakia, lens dislocation, glaucoma, joint stiffness, restricted articular movements and facial features. The anesthetic management of an 11-year-old-male patient with diagnosis of this syndrome is reported. PMID- 12648209 TI - Neuraxial complications after epidural and spinal anaesthesia. PMID- 12648210 TI - Extremely low oxygen tension in the rectal lumen of human subjects. PMID- 12648211 TI - Enrichment for living murine keratinocytes from the hair follicle bulge with the cell surface marker CD34. AB - It is widely believed that epithelial stem cells reside in the hair follicle bulge region. We investigated the hematopoietic stem and progenitor cell marker, CD34, as a potential marker of hair follicle bulge keratinocytes. Using a CD34 specific antibody, we identified intense membrane staining on keratinocytes in the bulge region of the mouse hair follicle. CD34 expression colocalized with both slowly cycling (label retaining) cells and keratin 15 expression. Live CD34+ keratinocytes were positively selected using antibodies to CD34 and alpha6 integrin in combination with fluorescent activated cell sorting. Sorted cells were analyzed for DNA content, and a staining profile was generated to confirm these cells as keratinocytes. CD34+ keratinocytes were predominantly in Go/G1, in contrast to CD34- cells, which had well defined G2/M and S phases. In addition, CD34+ keratinocytes were found to express alpha6 integrin more intensely than CD34- cells (p<0.05), identifying this population as an alpha6 integrin bright subset. When seeded at clonal density, CD34+ keratinocytes formed larger colonies than CD34- cells (p<0.05), indicating a higher proliferative potential. All flow sorted cells were positive for keratin 14 expression, and negative for keratin 1, loricrin, vimentin, and CD31. The majority of CD34+ cells (98%) were positive for keratin 6, establishing this population as basal keratinocytes of follicular origin. CD34 message was detected by reverse transcription polymerase chain reaction predominantly in the CD34+ keratinocytes, confirming specificity of the antibody. This work is the first to demonstrate that CD34 is a specific marker of bulge cell keratinocytes in the cutaneous epithelium. Furthermore, the use of this marker facilitates isolation of live epithelial cells with stem and progenitor cell characteristics, potentially providing a tool for the study of carcinogen target cells, gene therapy, and tissue engineering applications. PMID- 12648213 TI - Genetic evidence for a novel human desmosomal cadherin, desmoglein 4. AB - Desmosomes are essential adhesion structures in most epithelia that link the intermediate filament network of one cell to its neighbor, thereby forming a strong bond. The molecular components of desmosomes belong to the cadherin superfamily, the plakin family, and the armadillo repeat protein family. The desmosomal cadherins are calcium-dependent transmembrane adhesion molecules and comprise the desmogleins and desmocollins. To date, three human desmoglein isoforms have been characterized, namely desmogleins 1, 2, and 3 that are expressed in a tissue- and differentiation-specific manner. Here we have identified and characterized, at the genetic level, a novel human desmoglein cDNA sharing homology with desmogleins 1, 2, 3 and we name this desmoglein 4. The human desmoglein 4 cDNA (3.6 kb) contains an open reading frame of 3120 bp that encodes a precursor protein of 1040 amino acids. The predicted mature protein comprises 991 amino acids with a molecular weight of 107822 Da at pI 4.38. Human desmoglein 4 shares 41% identity with human desmoglein 1, 37% with human desmoglein 2, and 50% with human desmoglein 3. Analysis of the exon/intron organization of the human desmoglein 4 gene (DSG4) demonstrates that it is composed of 16 exons spanning approximately 37 kb of 18q12 and is situated between DSG1 and DSG3. We have demonstrated using RT-PCR on multiple tissue cDNA samples that desmoglein 4 has very specific tissue expression in salivary gland, testis, prostate, and skin. PMID- 12648212 TI - K6irs1, K6irs2, K6irs3, and K6irs4 represent the inner-root-sheath-specific type II epithelial keratins of the human hair follicle. AB - In this study we report on the cloning of two novel human type II keratin cDNAs, K6irs3 and K6irs4, which were specifically expressed in the inner root sheath of the hair follicle. Together with the genes of two previously described type II inner root sheath keratins, K6irs1 and K6irs2, the K6irs3 and K6irs4 genes were subclustered in the type II keratin/hair keratin gene domain on chromosome 12q13. Evolutionary tree analysis using all known type II epithelial and hair keratins revealed that the K6irs1-4 formed a branch separate from the other epithelial and hair keratins. RNA in situ hybridization and indirect immunofluorescence studies of human hair follicles, which also included the K6irs2 keratin, demonstrated that both K6irs2 and K6irs3 were specifically expressed in the inner root sheath cuticle, but showed a different onset of expression in this compartment. Whereas the K6irs3 expression began in the lowermost bulb region, that of K6irs2 was delayed up to the height of the apex of the dermal papilla. In contrast, the K6irs4 keratin was specifically expressed in the Huxley layer. Moreover, K6irs4 was ideally suited to further investigate the occurrence of Flugelzellen, i.e., Huxley cells, characterized by horizontal cell extensions that pass through the Henle layer, abut upon the companion layer, and form desmosomal connections with the surrounding cells. Previously, we detected Flugelzellen only in the region along the differentiated Henle layer. Using the Huxley-cell-specific K6irs4 antiserum, we now demonstrate this cell type to be clearly apposed to the entire Henle layer. We provide evidence that Flugelzellen penetrate the Henle layer actively and may play a role in conferring plasticity and resilience to the otherwise rigid upper Henle layer. PMID- 12648214 TI - Caveolin expression and localization in human keratinocytes suggest a role in lamellar granule biogenesis. AB - Lamellar granules are sphingolipid-enriched organelles, probably intimately related to the tubulo-vesicular elements of the trans-Golgi network, that deliver the precursors of stratum corneum barrier lipids to the extracellular compartment. Caveolins are cholesterol-binding scaffolding proteins that facilitate the assembly of cholesterol- and sphingolipid-enriched membrane domains known as caveolae. Similarities in the composition of lamellar granules and caveolae suggest that caveolins could be involved in lamellar granule assembly, trafficking, and/or function. In order to explore this relationship, we have examined the expression of caveolins in epidermis, keratinocyte cultures, and an isolated lamellar granule fraction using immunolabeling, immunoblotting, and northern blotting. Several antibodies show immunolocalization of caveolin-1 in the basal layer of human epidermis, with a decline in the suprabasal layers and a reemergence of expression at the stratum granulosum/stratum corneum junction. Two of three caveolin-2 antibodies show little basal staining, but strong signal throughout the rest of the epidermis, whereas a third shows a pattern like caveolin-1. An antibody against caveolin-3 shows a strong signal at the stratum granulosum/stratum corneum interface. Caveolins partially colocalize with glucocerebrosidase, an enzyme known to be critical for remodeling of extruded lamellar granule contents, with AE17, a previously described lamellar granule-associated antibody, and with glucosylceramides, a major lipid component of lamellar granules. Caveolin-1 protein is present in undifferentiated low calcium-grown keratinocyte cultures, decreases upon induction of differentiation, and then rises to levels above those seen in undifferentiated cultures, consistent with the immunofluorescence findings. Caveolin-1 mRNA expression parallels that of the protein. Caveolin-2 mRNA and protein expression were unchanged over the course of culture differentiation. Keratinocyte caveolin-1 mRNA expression is not induced by an increase in medium calcium level and is markedly reduced by phorbol-ester-mediated protein kinase C induction. Caveolin-1 is enriched in an isolated lamellar granule fraction that is also enriched, as we have previously described, in lysosomal acid lipase and glucocerebrosidase, and localizes to structures consistent with lamellar granules on immunoelectron microscopy. The differentiation-dependent expression of caveolin-1, the colocalization of caveolins with putative lamellar-granule-associated antigens, their enrichment in isolated lamellar granules, and their presence in lamellar granule-like structures on immunoelectron microscopy, along with their known structural role in the assembly of glycosphingolipid- and cholesterol-enriched domains in other cell types, suggest that caveolins may play a role in lamellar granule assembly, trafficking, and/or function. PMID- 12648215 TI - Function blocking autoantibodies against matrix metalloproteinase-1 in patients with systemic sclerosis. AB - Systemic sclerosis is characterized by fibrosis and systemic autoimmunity; however, roles of autoantibodies in the development of fibrosis remain unknown in systemic sclerosis. The net accumulation of extracellular matrix is dependent on the balance between the synthesis and degradation of extracellular matrix components, the latter process regulated by matrix metalloproteinases. Matrix metalloproteinase-1 (interstitial collagenase-1) can initiate degradation of collagen types I-III that are major extracellular matrix constituents in affected skin of systemic sclerosis. In this study, we tested the hypothesis that systemic autoimmunity in systemic sclerosis induced anti-matrix metalloproteinase-1 autoantibodies that inhibited matrix metallo-proteinase-1 activity, resulting in collagen accumulation. Enzyme-linked immunosorbent assay using human recombinant matrix metalloproteinase-1 revealed that IgG anti-matrix metalloproteinase-1 autoantibody levels were significantly elevated in sera from patients with systemic sclerosis, but not patients with active systemic lupus erythematosus or dermatomyositis, relative to normal controls. IgG anti-matrix metalloproteinase-1 autoantibody levels were significantly higher in patients with diffuse cutaneous systemic sclerosis than those found in patients with limited cutaneous systemic sclerosis. Furthermore, IgG anti-matrix metalloproteinase-1 antibody levels significantly correlated with the extent of fibrosis in the skin, lung, and renal blood vessels. The presence of IgG anti-matrix metalloproteinase-1 autoantibody in sera from systemic sclerosis patients was confirmed by immunoblotting analysis. Remarkably, IgG anti-matrix metalloproteinase-1 autoantibody in sera from systemic sclerosis patients inhibited matrix metalloproteinase-1 collagenase activity. Collectively, the results of this study suggest that anti-matrix metalloproteinase-1 autoantibody contributes to the development of fibrosis by inhibiting matrix metalloproteinase-1 collagenase activity and reducing the extracellular matrix turnover and suggest that the presence of anti-matrix metalloproteinase-1 autoantibody in systemic sclerosis is the link between systemic autoimmunity and fibrosis. PMID- 12648216 TI - Effect of smoking and sun on the aging skin. AB - Smoking and ultraviolet radiation are known to have a detrimental effect on human skin. Important characteristics of the aging skin are elastosis and telangiectasia. The purpose of the study was to assess the relative importance of age per se, and the detrimental effects caused by sun exposure and smoking on the development of cutaneous elastosis and telangiectasia in a well-defined group of individuals. We made use of 966 individuals who participated in a case-control study to investigate environmental and genetic risk factors for skin cancer. Exposure measurements for sunlight and smoking were collected and the amount of elastosis and telangiectasia in the face and neck was recorded according to a four-graded score varying from none to severe. Relative risks were estimated using exposure odds ratios from cross-tabulation and logistic regression. Multivariate logistic regression was used to adjust for potential confounders. Among both sexes a strong association was observed between increasing age, sun exposure, and amount of elastosis. The association between increasing age, sun exposure, and amount of telangiectasia was strong among men, but less apparent among women. Smoking was also associated with elastosis among both sexes, and with telangiectasia predominantly among men. Intrinsic differences between men and women (e.g., hormones) or behavior differences (e.g., more frequent use of creams and cosmetics among women) could account for this apparent difference in the occurrence of telangiectasia. In contrast to elastosis, telangiectasia may not be a good marker of the aging skin, specifically not in women. PMID- 12648217 TI - A cryotransmission electron microscopy study of skin barrier formation. AB - Direct visualization of the skin barrier formation process by cryotransmission electron microscopy of vitreous epidermal sections has been performed. The results obtained differ in many aspects from those of classical chemical fixation electron microscopy. Here we show that (i) a new "organelle or branched tubular structure" containing nonlamellar or partly lamellar material, and closely corresponding in numbers, dimensions, and localization to lamellar bodies of classical chemical fixation electron micrographs, and (ii) a new "ribosome complex-like structure", not preserved in classical electron micrographs, are omnipresent at apparent active sites of skin barrier formation. Evidence that skin barrier formation may not take place via extensive membrane fusion involving discrete lamellar bodies, but rather by a morphologically continuous membrane folding process are: (i) the often clearly nonlamellar content of structures corresponding to lamellar bodies with concomitant visualization of multilamellar membrane structures of the intercellular space; (ii) the "multifolded" appearance of the lipid matrix of the intercellular space; and (iii) the identification of extended "intracellular" multilamellar continuous structures with an optical density profile closely corresponding to that of the lipid matrix of the intercellular space. Based on the cryo-electron microscopic data presented in this study we propose that a membrane transition from cubic-like to multilamellar may take place already inside the "tubuloreticular cisternal membrane system" of upper granular cells recently reported by Elias et al. PMID- 12648218 TI - 1alpha,25-dihydroxyvitamin D3 stimulates activator protein 1 DNA-binding activity by a phosphatidylinositol 3-kinase/Ras/MEK/extracellular signal regulated kinase 1/2 and c-Jun N-terminal kinase 1-dependent increase in c-Fos, Fra1, and c-Jun expression in human keratinocytes. AB - 1alpha,25-Dihydroxyvitamin D3 added to human keratinocytes increases differentiation through an activation of the transcription factor activator protein 1. We have previously reported that the 1alpha,25-dihydroxyvitamin D3 induced increase of activator protein 1 DNA binding activity is mediated by a protein kinase C-independent mechanism. The purpose of this study was to investigate further the mechanisms by which 1alpha,25-dihydroxyvitamin D3 modulates activator protein 1 DNA binding activity in cultured normal human keratinocytes. Western blotting experiments revealed that 1alpha,25 dihydroxyvitamin D3 caused a rapid and transient activation of the mitogen activated protein kinases, extracellular signal regulated kinase 1/2 and c-Jun N terminal kinase 1. 1alpha,25-Dihydroxyvitamin D3 also enhanced the expression of the activator protein 1 subunits, c-Fos, Fra1, and c-Jun as determined by northern and western blotting. The 1alpha,25-dihydroxyvitamin D3-induced activator protein 1 DNA binding activity was completely blocked by the MEK inhibitor PD 98059 indicating that the MEK/extracellular signal regulated kinase pathway is involved in the activation of activator protein 1. Transfection experiments showed that 1alpha,25-dihydroxyvitamin D3 also increased the activator protein 1-dependent transactivation, which was completely blocked by expression of a dominant negative Ras, suggesting that the 1alpha,25 dihydroxyvitamin D3-induced activator protein 1 activity involves Ras-dependent signaling. Furthermore, preincubation of the keratinocytes with the specific phosphatidylinositol 3-kinase inhibitors, Wortmannin and LY294002, demonstrated that the 1alpha,25-dihydroxyvitamin D3-induced activation of extracellular signal regulated kinase 1/2 and c-Jun N-terminal kinase 1 required phosphatidylinositol 3-kinase activity. Finally, preincubation of keratinocytes with a polyclonal antibody against the membrane receptor annexin II, blocked the 1alpha,25 dihydroxyvitamin D3-induced activation of extracellular signal regulated kinase 1/2 and c-Jun N-terminal kinase 1. Taken together, our results indicate that 1alpha,25-dihydroxyvitamin D3, via binding to the membrane receptor annexin II, induces activation of the phos-phatidylinositol 3-kinase/Ras/MEK/extracellular signal regulated kinase 1/2 and c-Jun N-terminal kinase 1 signal transduction pathway resulting in increased expression of c-Fos, Fra1, and c-Jun, and subsequently increased activator protein 1 DNA binding activity and gene transcription. PMID- 12648220 TI - New acylceramide in native and reconstructed epidermis. AB - Culturing of normal human keratinocytes at the air-liquid interface results in the formation of fully differentiated epidermis under in vitro conditions. Although the reconstructed epidermis shows a close resemblance to native tissue, there are still some differences in the stratum corneum lipid profile and intercellular lipid organization. As ceramides belong to one of the major stratum corneum lipid classes, the aim of this study was to characterize this fraction in more detail. For this purpose, individual ceramide fractions were isolated by column chromatography and characterized by a combination of nuclear magnetic resonance spectroscopy, high-performance thin-layer chromatography, and gas chromatography. The results of this study show that in both the native and reconstructed human epidermis the extractable ceramide fraction contains, in addition to the well known acylceramides (EOS, EOH), a new acylceramide in which the omega-O-acylhydroxyacid is amide-linked to phytosphingosine (EOP). The same three sphingoid base moieties (S, P, H) are also found in ceramides with amide linked nonhydroxy and alpha-hydroxyacids. Whereas the same types of ceramides were present in both tissues, some differences in their fatty acid profiles have been found. In reconstructed epidermis the content of linoleic acid in all three acylceramides fraction was significantly lower; the ceramide(NS) fraction was enriched in short fatty acids and the ceramide(AS) fraction was enriched in long chain alpha-hydroxyacids. These differences together with a lower content of free fatty acids may explain the differences between native and reconstructed tissue in stratum corneum lipid organization observed earlier by X-ray diffraction. PMID- 12648219 TI - Suppressor of cytokine signaling 1/JAB and suppressor of cytokine signaling 3/cytokine-inducible SH2 containing protein 3 negatively regulate the signal transducers and activators of transcription signaling pathway in normal human epidermal keratinocytes. AB - The suppressor of cytokine signaling/cytokine-inducible SH2 containing proteins are cytokine inducible and are negative regulators of the signal transducers and activators of the transcription signaling pathway. We investigated the mechanism regulating signal transducers and activators of transcription and the suppressor of cytokine signaling/cytokine-inducible SH2 containing protein family in keratinocytes, one of the major target cells for cytokines. Suppressor of cytokine signaling 1 mRNA was upregulated 3 h post-interferon gamma, and a 8.1 fold increase in the suppressor of cytokine signaling 1 mRNA occurred 48 h post interferon gamma. The suppressor of cytokine signaling 3 mRNA was also upregulated from 1 h post-interferon gamma, and a 6.7-fold increase in the suppressor of cytokine signaling 3/cytokine-inducible SH2 containing protein 3 mRNA occurred between 6 and 12 h post-interferon gamma. Interleukin-6 exposure for 1 h enhanced the expression of the suppressor of cytokine signaling 3/cytokine-inducible SH2 containing protein 3 mRNA, but the suppressor of cytokine signaling 1/JAB mRNA was not induced by interleukin-6. Interleukin-4 upregulated the suppressor of cytokine signaling 1/JAB and cytokine-inducible SH2 containing protein 1 mRNA, with 3.4-fold and 5.1-fold increases in mRNA observed at 1 h post-interleukin-4, respectively. In contrast, epidermal growth factor, which phosphorylates signal transducers and activators of transcription 3, did not influence the level of the suppressor of cytokine signaling/cytokine inducible SH2 containing protein family mRNA expression. Transfection of an adenovirus vector expressing the suppressor of cytokine signaling 1/JAB completely inhibited interferon gamma-dependent signal transducers and activators of transcription 1 phosphorylation and interleukin-4-dependent signal transducers and activators of transcription 6 phosphorylation. Transfection of adenovirus vector expressing the suppressor of cytokine signaling 1/JAB did not inhibit interleukin-6-dependent signal transducers and activators of transcription 3 phosphorylation-several reports show that the suppressor of cytokine signaling 1/JAB is a potent inhibitor of signal transducers and activators of transcription 3 signaling in the myeloid leukemia M1 cell. Transfection of the adenovirus vector expressing suppressor of cytokine signaling 3/cytokine-inducible SH2 containing protein 3 completely inhibited interleukin-6-dependent signal transducers and activators of transcription 3 phosphorylation and partially inhibited interferon gamma-dependent signal transducers and activators of transcription 1 phosphorylation. Transfection of the adenovirus vector expressing suppressor of cytokine signaling 3/cytokine-inducible SH2 containing protein 3, however, did not inhibit interleukin-4-dependent signal transducers and activators of transcription 6 phosphorylation. Transfection of the adenovirus vector expressing cytokine-inducible SH2 containing protein 1 had no effect on signal transducers and activators of transcription 1, 3, and 6 signaling in normal keratinocytes. Therefore, the relationship between signal transducers and activators of transcription and suppressor of cytokine signaling is unique in the keratinocytes, and the suppressor of cytokine signaling regulates cytokine signals in these cells. PMID- 12648221 TI - Bovine-serum-albumin-containing receptor phase better predicts transdermal absorption parameters for lipophilic compounds. AB - Based on the hypothesis that limited receptor solubility of lipophilic compounds may result in lower observed permeability parameters, the aim of this study was to determine the in vitro human epidermal permeability coefficients and membrane retention of a series of aliphatic alcohols (C1-C10, log p-0.72 to 4.06) using two different receptor solutions (water and 4% bovine serum albumin in phosphate buffered saline). Aqueous solutions of radiolabeled alcohols were dosed into the stratum corneum side of membranes mounted in side-by-side glass diffusion cells. Appearance of alcohol in the receptor compartment filled with either of the two solutions was monitored over a 7 h period when both stratum corneum (assessed by tape stripping) and the remaining epidermis levels of radioactivity were determined. In a separate study the degree of binding of alcohols to 4% bovine serum albumin was determined. The data showed increased receptor phase solubility in the bovine serum albumin solution and higher permeability coefficients for the more lipophilic alcohols in the series. No changes were seen in the partitioning of the alcohols from the vehicle into either the stratum corneum or tape-stripped epidermis with the two receptor phases; however, a decrease in the amount of the more lipophilic alcohols partitioning into the water receptor phase from the tape stripped epidermis was observed. We conclude that bovine serum albumin receptor phase allows better estimation of real permeability parameters for lipophilic compounds due to its increased solubility capacity and we question whether permeability parameters for lipophilic solutes from older data sets based on aqueous receptor phases are completely reliable. PMID- 12648222 TI - Analysis of proteins with caseinolytic activity in a human stratum corneum extract revealed a yet unidentified cysteine protease and identified the so called "stratum corneum thiol protease" as cathepsin l2. AB - Desquamation is described as a protease-dependent phenomenon where serine proteases with a basic pH optimum play a key role. Recently proteases with an acidic pH optimum were identified in the stratumcorneum and associated with desquamation, e.g., cathepsin D and the stratum corneum thiol protease. The purpose of this study was to investigate if human stratum corneum contains proteases different from the above, exhibiting similar properties. After gel filtration, we identified four distinct proteolytic activities in a human stratum corneum extract, a cathepsin-E-like activity (80 kDa), a cathepsin-D activity (40 kDa), a yet unknown cathepsin-L-like form (28 kDa) exhibiting the highest caseinolytic activity, and a chymotrypsin-like protein (24 kDa) containing the acidic activity of the well described stratum corneum chymotryptic enzyme. We named the new 28 kDa protease stratum corneum cathepsin-L-like enzyme. Characterization of stratum corneum cathepsin-L-like enzyme provided clear evidence that this new protease, despite its membership to the cathepsin-L-like family, is distinct from cathepsin L and from the recently described stratum corneum thiol protease. Its ability to hydrolyze corneodesmosin, a marker of corneocyte cohesion, was in favor of a role of stratum corneum cathepsin-L-like enzyme in the desquamation process. A more detailed analysis did not allow us to identify stratum corneum cathepsin-L-like enzyme at the molecular level but revealed that stratum corneum thiol protease is identical with the recently described cathepsin L2 protease. Reverse transcription polymerase chain reaction studies and the use of a specific antibody revealed that, in contrast to earlier reports, expression of stratum corneum thiol protease in human epidermis is not related to keratinocyte differentiation. Our results indicate that the stratum corneum thiol protease is probably expressed as a pro-enzyme in the lower layers of the epidermis and in part activated by a yet unidentified mechanism in the upper layers during keratinocyte differentiation. PMID- 12648223 TI - Genetic heterogeneity in erythrokeratodermia variabilis: novel mutations in the connexin gene GJB4 (Cx30.3) and genotype-phenotype correlations. AB - Erythrokeratodermia variabilis is an autosomal dominant genodermatosis characterized by persistent plaque-like or generalized hyperkeratosis and transient red patches of variable size, shape, and location. The disorder maps to a cluster of connexin genes on chromosome 1p34-p35.1 and, in a subset of families, results from mutations in the gene GJB3 encoding the gap junction protein connexin-31 (Cx31). A recent report suggested the involvement of another connexin gene (GJB4) in the etiology of erythrokeratodermia variabilis. In this study, we sequenced the coding region of GJB4 in 13 unrelated erythrokeratodermia variabilis families without detectable mutations in GJB3. Mutation analysis revealed six distinct missense mutations in five families and a sporadic case of erythrokeratodermia variabilis, all of which were not found in controls. Mutation G12D, identified in an extended Dutch family, lies in the predicted amino terminus and may interfere with the flexibility of this domain, connexin selectivity, or gating polarity of gap junction channels. Other mutations (R22H, T85P, F137L, F189Y) were located in the transmembrane domains of Cx30.3, and are predicted to hinder regulation of voltage gating or alter the kinetics of channel closure. Affected individuals of two unrelated families harbored point mutations leading to amino acid substitution F137L, which was also reported in GJB3, yet the extent and severity of hyperkeratosis was milder compared to the corresponding mutation in GJB3. Two mutations (T85P, F137L) were associated with the occurrence of rapidly changing erythematous patches with prominent, circinate, or gyrate borders in affected children but not in adults, supporting the notion that this feature is specific to Cx30.3 defects. Nevertheless, we observed highly variable intrafamilial phenotypes, suggesting the strong influence of modifying genetic and epigenetic factors. In addition to pathogenic mutations, we identified several missense mutations and a 4 bp deletion within the GJB4 coding region, which might represent either inconsequential polymorphisms or recessive mutations. In conclusion, our results demonstrate genetic heterogeneity in erythrokeratodermia variabilis, and emphasize that intercellular communication mediated by both Cx31 and Cx30.3 is crucial for epidermal differentiation. PMID- 12648225 TI - Association of TAP and HLA-DM genes with psoriasis in Koreans. AB - To investigate the possible involvement of antigen-processing genes in the pathogenesis of psoriasis, we analyzed the polymorphisms of the TAP1, TAP2, LMP2, LMP7, DMA, and DMB genes in 98 Korean psoriasis patients and compared them with 184 healthy controls. The frequencies of TAP2*B/B [relative risk (RR)=3.6, p<0.0002] and TAP2*B (RR=1.7, p<0.05) were significantly increased, but TAP1*B (RR=0.3, p<0.002) and TAP2*A (RR=0.6, p<0.03) were significantly decreased, in the patients compared to the controls. We performed further analysis on the TAP1 and TAP2 single nucleotide polymorphisms and found significant differences between the patients and controls in TAP1 single nucleotide polymorphism at position 637 and in TAP2 at 665. In HLA-DM, DMA*0102 (RR=2.5, p<0.0003) was significantly increased, but DMA*0101/0101 (RR=0.4, p<0.0004) and DMB*0103/0103 (RR= 0.3, p<0.005) were significantly decreased in the patients compared to the controls. The TAP and HLA-DM alleles were also analyzed according to the age of onset of psoriasis in the patients (types I and II). It was found that the HLA-DM alleles showed a greater association in type I than type II patients. An analysis of the linkage disequilibrium and stratification also indicated that the alleles of TAP and HLA-DM might be independently associated with HLA-Cw*0602 in psoriasis patients. The stratification analysis between DMA*0101/0101 and DMB*0103/0103 showed that a certain factor, controlled by a gene located between DMA and DMB, might provide strong protection against psoriasis, independently of Cw*0602, in our Korean population. In conclusion, our data suggest that the TAP and HLA-DM alleles could lead to genetic susceptibility toward psoriasis in Koreans. PMID- 12648224 TI - Suppression of the immune response against exogenous desmoglein 3 in desmoglein 3 knockout mice: an implication for gene therapy. AB - Gene therapies for recessive genetic diseases may provoke unwanted immune responses against the introduced gene product because patients, especially those with null mutation of a certain protein, have no tolerance for the protein of interest. This study used desmoglein 3 knockout (Dsg3-/-) mice as a disease model for a genetic defect in DSG3, to investigate whether nonviral gene therapy induces an immune response against Dsg3 and whether the reaction against Dsg3 can be prevented. When mouse Dsg3 cDNA was injected in the skin of Dsg3-/- mice, 50% of treated Dsg3-/- mice developed anti-Dsg3 IgG, which can bind native Dsg3 in vivo. To prevent this response, we used an anti-CD40L monoclonal antibody, MR1, which blocks the costimulatory interaction between CD40 and CD40L. To evaluate the effect of MR1, we grafted Dsg3+/+skin on Dsg3-/- mice, to mimic stable gene transfer of Dsg3. After skin grafting, all the recipient Dsg3-/- mice were treated with either MR1 (n=8) or control hamster IgG (n=8). All of the control IgG-treated mice developed circulating anti-Dsg3 IgG about 2 wk after grafting, and IgG deposition was observed on the surfaces of keratinocytes in the grafted Dsg3+/+skin. Such anti-Dsg3 IgG production was significantly prevented, however, when the recipient mice were treated with MR1. These findings suggested that gene therapies for recessive diseases may provoke an immune response against the transgene product, and that the CD40-CD40L interaction might be a reasonable target for effective prevention of such undesirable immune responses, leading, in turn, to a successful gene therapy. PMID- 12648226 TI - Epidermolytic hyperkeratosis and epidermolysis bullosa simplex caused by frameshift mutations altering the v2 tail domains of keratin 1 and keratin 5. AB - The cytoskeleton of epithelial cells is formed by heteropolymeric keratin proteins characterized by a central alpha-helical rod flanked by nonhelical head and tail domains of variable sequence. Most mutations described in 18 distinct keratins disrupt highly conserved regions at the boundaries of the rod, which have been recognized as zones of overlap during keratin alignment and assembly into intermediate filaments. We recently reported the first mutation located in a keratin tail domain (V2) in ichthyosis hystrix Curth-Macklin. In this study, we report two novel frameshift mutations that are predicted to alter the tail of keratin 1 or keratin 5, leading to an atypical form of epidermolytic hyperkeratosis and a mild form of epidermolysis bullosa simplex, respectively. Mutation analysis of the patient with epidermolytic hyperkeratosis revealed a de novo heterozygous nucleotide insertion (1752insG) in exon 9 of KRT1, predicted to result in an aberrant 69 residue keratin 1 tail. In the patient with mild epidermolysis bullosa simplex, we identified a single nucleotide deletion (1635delG) in exon 9 of KRT5 leading to frameshift and translation of an abnormal V2 domain, 35 amino acids longer than the native keratin 5 tail. Our results, together with previous observations, establish the existence of a subgroup of keratin disorders due to frameshift mutations altering the keratin tail domains that are characterized by phenotypic heterogeneity. PMID- 12648227 TI - Genetic analysis of PSORS1 distinguishes guttate psoriasis and palmoplantar pustulosis. AB - The PSORS1 locus in the major histocompatibility complex region is the major genetic determinant for psoriasis vulgaris. Within the PSORS1 region reside at least three potential candidate genes for psoriasis susceptibility. Specific allelic variants of the genes HLA-Cw*6, HCR*WWCC, and CDSN*5 are strongly associated with psoriasis vulgaris and are in strong linkage disequilibrium with each other. We have genotyped the three psoriasis vulgaris susceptibility alleles of the PSORS1 locus in two clinical variants of psoriasis (guttate psoriasis and palmoplantar pustulosis) to study whether PSORS1 is also involved in the pathogenesis of these variants. We also asked whether these two clinical subgroups could help us to distinguish the causative gene within the high-risk PSORS1 haplotype. The association of guttate psoriasis with the three PSORS1 susceptibility alleles was similar and even stronger than seen with psoriasis vulgaris. Palmoplantar pustulosis, however, did not show association with any of the three candidate genes at this locus. Finally, no correlation with the age of onset for disease was observed. Our results show conclusively that psoriasis vulgaris and guttate psoriasis have a similar genetic basis for their association to PSORS1, whereas palmoplantar pustulosis appears to be a distinct disorder. PMID- 12648228 TI - Epicutaneous natural rubber latex sensitization induces T helper 2-type dermatitis and strong prohevein-specific IgE response. AB - In addition to immediate type I allergy symptoms, natural rubber latex allergy may manifest as protein contact dermatitis on the hands of health-care workers and other natural rubber latex glove users. We examined whether repeated application of natural rubber latex on mouse skin causes sensitization to natural rubber latex and dermatitis. Epicutaneous sensitization with natural rubber latex produced a significant influx of mononuclear cells, CD4+ CD3+ cells, and eosinophils to the sensitized skin sites. The number of degranulated mast cells in natural rubber latex-sensitized skin sites was significantly higher compared with control sites treated with phosphate-buffered saline. The expression of interleukin-1beta and interleukin-4 mRNA was markedly increased in natural rubber latex-sensitized skin sites. Moreover, significant increases in the mRNA expression of chemokines CCL2 (monocyte chemoattractant protein-1), CCL11 (eotaxin-1), CCL3 (macrophage inflammatory protein-1alpha), and CCL4 (macrophage inflammatory protein-1beta) were found. In addition to the cutaneous inflammatory response, epicutaneous sensitization with natural rubber latex induced a striking increase in the total and specific immunoglobulin E levels but not in the immunoglobulin G2a levels. Intraperitoneal immunization with natural rubber latex induced a strong natural rubber latex-specific immunoglobulin G2a response, but only a weak immunoglobulin E response. We also studied the role of two major natural rubber latex allergens, the highly hydrophilic prohevein and the hydrophobic rubber elongation factor. Cutaneous application of natural rubber latex elicited a strong immunoglobulin E response against prohevein, but not against rubber elongation factor. On the contrary, intraperitoneal immunization with natural rubber latex elicited strong immunoglobulin G2a production to rubber elongation factor but not to prohevein. These results demonstrate that epicutaneous sensitization with natural rubber latex induces T helper 2-dominated dermal inflammation and strong immunoglobulin E response in this murine model of natural rubber latex induced protein contact dermatitis. Epicutaneous sensitization to natural rubber latex proteins eluting from latex gloves may therefore contribute to the development of hand dermatitis and also natural rubber latex-specific immunoglobulin E antibodies. PMID- 12648229 TI - Afferent and efferent phases of allergic contact dermatitis (ACD) can be induced after a single skin contact with haptens: evidence using a mouse model of primary ACD. AB - Allergic contact dermatitis is a T cell-mediated delayed type hypersensitivity reaction that occurs upon hapten challenge in sensitized individuals. The inflammatory response in classical allergic contact dermatitis requires both a sensitization phase and an elicitation phase responsible for the recruitment and activation of specific T cells at the site of hapten skin challenge. Conversely, previously unsensitized patients may develop a "primary allergic contact dermatitis" after the first skin contact with potent contact sensitizers leading to a skin inflammation with all the features of classical allergic contact dermatitis. In this study we used an experimental murine model, referred to as contact hypersensitivity, to study the pathophysiology of primary allergic contact dermatitis and its relationship to classical allergic contact dermatitis. We show that one epicutaneous application of a nonirritant dose of hapten (2,4 dini-trofluorobenzene, fluorescein isothiocyanate) was sufficient to induce an optimal allergic contact dermatitis reaction at the site of primary contact with the hapten without subsequent challenge. As in classical allergic contact dermatitis, the skin inflammation in primary allergic contact dermatitis was mediated by interferon-gamma producing, CD8+ effector T cells that were induced in the draining lymph nodes at day 5 postsensitization and downregulated by CD4+ T cells. Reverse transcription-polymerase chain reaction analysis revealed that the primary allergic contact dermatitis reaction was mediated by a recruitment of CD8+ T cells at the sensitization skin site at day 6 postsensitization. Analysis of the fate of the hapten fluorescein isothiocyanate applied once on the skin revealed its persistence in the epidermis for up to 14 d after skin painting. These results suggest that the development of primary allergic contact dermatitis (i.e., without secondary challenge) is associated with persistence of the hapten in the skin, which allows the recruitment and activation of CD8+ T cells at the site of the single hapten application. PMID- 12648231 TI - Influence of hydration on dihydroxyacetone-induced pigmentation of stratum corneum. AB - Dihydroxyacetone, the browning ingredient in sunless tanning formulations, reacts with amino acids in the outer stratum corneum to form a mixture of high molecular weight pigments. Our initial observations indicated that high hydration of dihydroxyacetone-treated skin completely inhibited development of pigmentation. To investigate the mechanism underlying this effect, studies were carried out in isolated murine epidermis, polyvinyl alcohol/lysine films, and lysine in glycerol/water solvent. Murine epidermis treated with dihydroxyacetone showed a biphasic dependence on relative humidity: maximum pigmentation developed at 84% relative humidity and minimum pigmentation at 0% and 100% relative humidity. Filaggrin proteolysis, which shows a similar dependence on relative humidity and provides free amino acids in the outer stratum corneum, did not account for the relative humidity dependence of dihydroxyacetone pigmentation. A similar biphasic pigmentation response was obtained when polyvinyl alcohol film containing lysine was treated with dihydroxyacetone and incubated at various relative humidities, indicating that the structure of the stratum corneum was not a major factor. To remove the influence of the matrix, the reaction of dihydroxyacetone with lysine was followed at varying concentrations of water in mixed glycerol/buffer solvent. Again, greater pigment formation was found at an intermediate level of water (6% vol/vol) and little pigmentation at 0% and 100% water content. These results are consistent with a requirement for water at low relative humidity, which facilitates formation of free amine groups needed for the initial reaction with dihydroxyacetone, and with inhibition of the dehydration reactions by water through the law of mass action at high relative humidity. PMID- 12648230 TI - Radiosensitization of human melanoma cells by ribozyme-mediated inhibition of survivin expression. AB - Survivin is a structurally unique member of the inhibitors of apoptosis protein family and is involved in the control of cell division and inhibition of apoptosis. The notion that survivin is overexpressed in most human tumors but absent in normal adult tissues with only a few exceptions has led to the proposal of survivin as a promising therapeutic target for novel anticancer therapies. In this context, we generated a hammerhead ribozyme targeting the 3' end of the CUA110 triplet in the survivin mRNA. Two human melanoma cell lines (JR8 and M14) overexpressing survivin were stably transfected with the pRc/CMV vector carrying the ribozyme sequence. Two polyclonal cell populations proven to endogenously express ribozyme and characterized by a markedly lower survivin protein level ( 60% and -50%, respectively) than JR8 and M14 parental cells were selected for the study. Ribozyme-expressing cells showed a significantly (p<0.01) increased sensitivity to gamma-irradiation (as detected by clonogenic cell survival) compared to JR8 and M14 cells. Moreover, in the JR8 cell line, the extent of radiation-induced apoptosis (in terms of percentage of apoptotic nuclei in cells stained with propidium iodide and level of caspase-3 catalytic activity) was markedly greater in ribozyme-expressing cells than in parental cells. These results demonstrate for the first time that attenuation of survivin expression renders human melanoma cells more susceptible to gamma-irradiation. PMID- 12648232 TI - De novo ceramide synthesis participates in the ultraviolet B irradiation-induced apoptosis in undifferentiated cultured human keratinocytes. AB - Ultraviolet irradiation is a major environmental cause of skin cancers, whereas ultraviolet-induced DNA repair and apoptosis are defense mechanisms that rescue and/or protect keratinocytes from this risk. Multiple pathways are involved in ultraviolet-induced keratinocyte apoptosis, including activation of p38-mitogen activated protein kinase, protein kinase C, and CD95, each of which are associated with caspase activation. Alternatively, ceramides could serve as ultraviolet-induced, second messenger lipids, because they induce cell cycle arrest and apoptosis in a variety of cell types, including keratinocytes. We investigated the role of ceramide versus caspase, and the responsible pathway for ceramide generation in ultraviolet B-induced apoptosis of cultured normal human keratinocytes maintained in low calcium (0.07 mm) medium. Ultraviolet B (40 mJ per cm2) significantly inhibited cultured normal human keratinocyte proliferation, assessed as [3H-methyl]thymidine-thymidine incorporation into DNA, 2 h after irradiation. Terminal nick deoxynucleotide end-labeling-positive apoptotic cells (14.8% at 24 h and 34.4% at 48 h) and trypan blue-positive apoptotic cells (8.4% at 24 h and 28.6% at 48 h) became evident in a time dependent manner after ultraviolet B irradiation, in parallel with activation of caspase-3. The ceramide content of irradiated cultured normal human keratinocytes increased significantly by 8 h, whereas glucosylceramide only modestly increased, and sphingomyelin content remained unaltered. Metabolic studies with radiolabeled serine, palmitic acid, and phosphorylcholine revealed that the ultraviolet B induced increase in ceramide results primarily from increased de novo synthesis rather than accelerated sphingomyelin hydrolysis. Increased ceramide synthesis, in turn, could be attributed to increased activity of ceramide synthase (i.e., 1.7-fold increase 8 h after ultraviolet B irradiation), whereas serine palmitoyltransferase activity did not change. Both fumonisin B1, an inhibitor of ceramide synthase, and ISP-1, myriocin an inhibitor of serine palmitoyltransferase, significantly attenuated the ultraviolet B-induced apoptosis in a caspase-3-independent fashion, whereas co-incubation with a caspase-3 inhibitor (Ac-DEVD-chloromethyl-ketone) further attenuated the ultraviolet B-induced apoptosis. Thus, increased de novo ceramide synthesis signals ultraviolet B-induced apoptosis, by a pathway independent of, but in concert with, caspase-3 activation. PMID- 12648233 TI - T cell receptor-gamma gene analysis of CD30+ large atypical individual cells in CD30+ large primary cutaneous T cell lymphomas. AB - The hallmark of primary cutaneous CD30+ large T cell lymphoma are large lymphoid tumor cells, at least 75% of which, by definition, must be positive for CD30. The relatively benign clinical course of this lymphoma type has been explained with CD30-induced apoptosis, on the assumption that expression of CD30 defines the tumor clone; however, this hypothesis has not been tested on the molecular level to date. In this study we analyzed CD30+ cells in four patients with primary cutaneous CD30+ large T cell lymphoma by single cell polymerase chain reaction of T cell receptor-gamma genes followed by sequencing. Here, we demonstrate that most of the large CD30+ atypical cells possessed identical T cell receptor-gamma gene rearrangements, indicative of clonal proliferation. Nevertheless, polyclonally rearranged T cells were present in all CD30+ samples studied. In addition, one patient showed a second clone in a separate biopsy and three of four patients showed chromosomal imbalances as revealed by comparative genomic hybridization. Taken together, our data suggest that the CD30+ population in primary cutaneous CD30+ large T cell lymphoma indeed contains the tumor clone, thus providing molecular support for a link between clinical course and CD30 related signaling. Importantly, however, CD30 expression does not define the tumor clone as bystander T cells, as well as occasional additional clones, are also present in this population. PMID- 12648234 TI - High frequency of ultraviolet mutations at the INK4a-ARF locus in squamous cell carcinomas from psoralen-plus-ultraviolet-A-treated psoriasis patients. AB - Squamous cell carcinomas in psoralen-plus-ultraviolet A (PUVA) treated patients frequently exhibit p53 tumor suppressor genes and Ha-ras protooncogenes that are mutated at dipyrimidine sites and carry the ultraviolet fingerprint (i.e., C-to-T or CC-to-TT transitions). To further broaden the knowledge of genetic mutations in PUVA-associated skin cancer, we used DNA sequencing analysis to study the mutational spectrum of the INK4a-ARF locus in 26 squamous cell carcinomas from 11 long-term PUVA-treated psoriasis patients and classified the mutations by origin (ultraviolet, ultraviolet and/or PUVA, or other). Nineteen INK4a-ARF missense/nonsense mutations were found in exons 1alpha, 1beta, and 2 in 11 of 26 squamous cell carcinomas (42%) from seven of 11 patients (64%). Eleven mutations (58%) were of the ultraviolet type; three (16%) were of the ultraviolet and/or PUVA type (i.e., C-to-T transitions at dipyrimidine sites opposite a 5'TpG sequence, a potential psoralen binding site); and five (26%) were of other type. Interestingly, 10 of 11 patients (91%) showed intron polymorphism C500G at the 3' untranslated region of exon 3. These data indicate that (i) INK4a-ARF mutations frequently occur in PUVA-associated squamous cell carcinomas; (ii) ultraviolet B radiation is the major cause of these mutations; and (iii) PUVA itself may play no direct role in development of most INK4a-ARF mutations. PMID- 12648235 TI - Stroma formation and angiogenesis by overexpression of growth factors, cytokines, and proteolytic enzymes in human skin grafted to SCID mice. AB - Reorganization of skin during wound healing, inflammatory disorders, or cancer growth is the result of expression changes of multiple genes associated with tissue morphogenesis. We wanted to identify proteins involved in skin remodeling and select those that may be targeted for agonistic or antagonist therapeutic approaches in various disease processes. Full-thickness human skin was grafted to severe combined immunodeficient mice and injected intradermally with 38 different adenoviral vectors inserted with 37 different genes coding for growth factors, cytokines, proteolytic enzymes and their inhibitors, adhesion receptors, oncogenes, and tumor suppressor genes. Responses were characterized for infiltration of inflammatory cells, vascular density, matrix formation, fibroblast-like cell proliferation, and epidermal hyperplasia. Of the 17 growth factor vectors, 16 induced histological changes in human skin. Members of the VEGF and angiopoietin families induced neovascularization. PDGFs and TGF-betas stimulated connective tissue formation, and the chemokines IL-8 and MCP-1 attracted inflammatory neutrophils and monocytes, respectively. The serine protease uPA induced a vascular response similar to that of VEGF. Vectors with adhesion receptors, oncogenes and tumor suppressor genes had, with few exceptions, little effects on skin architecture. The overall results suggest that adenoviral vectors can effectively remodel the architecture of human skin for studies in morphogenesis, inflammatory skin disorders, wound healing, and cancer development. PMID- 12648236 TI - Sphingosine-1-phosphate and its potentially paradoxical effects on critical parameters of cutaneous wound healing. AB - The sphingolipid metabolite sphingosine-1-phosphate has emerged as a new bioactive molecule involved in the regulation of cell growth, differentiation, survival, and chemotaxis as well as angiogenesis and embryogenesis. These effects are mediated either via G-protein-coupled receptors or through intracellular actions. The most prominent sources of sphingosine-1-phosphate are human platelets suggesting its potential role in wound healing. In agreement with a positive function on reconstruction of wounded skin, we identified sphingosine-1 phosphate as a potent chemoattractant for keratinocytes as well as an activator of extracellular matrix production by fibroblasts. An unexpected finding is a strong cell growth arrest of keratinocytes after exposure to sphingosine-1 phosphate, as keratinocyte proliferation is critical for re-epithelialization of the wound. Most interestingly, the anti-proliferative effect of sphingosine-1 phosphate is not a result of cytotoxicity or apoptosis as sphingosine-1-phosphate even protects these cells from programmed cell death. Moreover, sphingosine-1 phosphate enhances differentiation of keratinocytes. To investigate further by which signaling pathway cell growth inhibition is mediated expression of the mRNA of all sphingosine-1-phosphate receptors (S1P1-5) was identified. 1 (Edg 1), 2 (Edg 5), 3 (Edg 3), 4 (Edg 6), and 5 (Edg 8) mRNA in keratinocytes was identified. As demonstrated in guanosine 5-[gamma-35S] triphosphate-gammaS binding assays, these G-protein-coupled receptors are functional at nanomolar concentrations. As the anti-proliferative effect of sphingosine-1-phosphate is only partially inhibited in the presence of pertussis toxin, it was investigated if intracellular actions are also involved. Microinjections of sphingosine-1 phosphate in keratinocytes also reduce proliferation suggesting that both sphingosine-1-phosphate receptors as well as intracellular actions mediate sphingosine-1-phosphate- induced cell growth arrest. PMID- 12648238 TI - The crucial role of TGF-beta in the age-related alterations induced by ultraviolet A irradiation. PMID- 12648237 TI - Autoantibody in mucous membrane pemphigoid binds to an intracellular epitope on human beta4 integrin and causes basement membrane zone separation in oral mucosa in an organ culture model. PMID- 12648239 TI - Greater diversity of desmosomal cadherins. PMID- 12648241 TI - In this issue: progress towards a more complete understanding of two important immune diseases involving skin. PMID- 12648242 TI - The virtue of tolerance. PMID- 12648243 TI - Paradoxical effects of sphingosine-1-phosphate. PMID- 12648244 TI - Caveolins in epidermal lamellar bodies: skin is an interactive interface, not an inflexible barrier. PMID- 12648245 TI - Genotyping psoriasis. PMID- 12648246 TI - Variation in predictors of primary care career choice by year and stage of training. AB - CONTEXT: It is not known whether factors associated with primary care career choice affect trainees differently at different times or stages of medical education. OBJECTIVE: To examine how role models, encouragement, and personal characteristics affect career choice at different stages (medical school vs residency) and periods (1994 vs 1997) of training. DESIGN: A split-panel design with 2 cross-sectional telephone surveys and a panel survey in 1994 and 1997. PARTICIPANTS: A national probability sample of fourth-year students (307 in 1994, 219 in 1997), 645 second-year residents in 1994, and 494 third-year residents in 1997. Of the fourth-year students interviewed in 1994, 241 (78.5%) were re interviewed as third-year residents in 1997. MAIN OUTCOME MEASURE: Primary care (general internal medicine, general pediatrics, or family medicine) career choice. RESULTS: Having a primary care role model was a stronger predictor of primary care career choice for residents (odds ratio [OR], 18.0; 95% confidence interval [95% CI], 11.2 to 28.8 in 1994; OR, 43.7; 95% CI, 24.4 to 78.3 in 1997) than for students (OR, 6.5; 95% CI, 4.3 to 10.2; no variation by year). Likewise, peer encouragement was more predictive for residents (OR, 5.4; 95% CI, 3.3 to 8.9 in 1994; OR, 16.6; 95% CI; 9.7 to 28.4 in 1997) than for students (OR, 2.1; 95% CI, 1.3 to 3.2; no variation by year). Orientation to the emotional aspects of care was consistently associated with primary care career choice across stages and years of training. CONCLUSIONS: The effect of peer encouragement and role models on career choice differed for students and residents and, in the case of residents, by year of training, suggesting that interventions to increase the primary care workforce should be tailored to stage of training. PMID- 12648247 TI - House staff nonverbal communication skills and standardized patient satisfaction. AB - OBJECTIVE: To examine the association of physician nonverbal communication with standardized patient (SP) satisfaction in the context of the "quality" of the interview (i.e., information provided and collected, communication skills). DESIGN: Observational. SETTING: One university-based internal medicine residency program. PARTICIPANTS: Fifty-nine internal medicine residents. INTERVIEWING: The 59 residents were recruited to participate in 3 SP encounters. The scenarios included: 1) a straightforward, primarily "medical" problem (chest pain); 2) a patient with more psychosocial overlay (a depressed patient with a history of sexual abuse); and 3) a counseling encounter (HIV risk factor reduction counseling). Trained SPs rated physician nonverbal behaviors (body lean, open versus closed body posture, eye contact, smiling, tone of voice, nod, facial expressivity) in the 3 encounters. Multiple regression approaches were used to investigate the association of physician nonverbal behavior with patient satisfaction in the context of the "quality" of the interview (SP checklist performance, measures of verbal communication skills), controlling for physician characteristics (gender, postgraduate year). RESULTS: Nonverbal communication skills was an independent predictor of standardized patient satisfaction for all 3 patient stations. The effect sizes were substantial, with nonverbal communication predicting 32% of the variance in patient satisfaction for the chest pain station, 23% of the variance for the depression-sexual abuse station, and 19% of the variance for the HIV counseling station. CONCLUSION: Better nonverbal communication skills are associated with significantly greater patient satisfaction in a variety of different types of clinical encounters with standardized patients. Formal instruction in nonverbal communication may be an important addition to residency. PMID- 12648248 TI - Patient satisfaction in women's clinics versus traditional primary care clinics in the Veterans Administration. AB - OBJECTIVE: To compare patient satisfaction in women's clinics (WCs) versus traditional primary care clinics (TCs). DESIGN: Anonymous, cross-sectional mailed survey. SETTING: Eight Department of Veterans Affairs (VA) medical centers in 3 states. PATIENTS: A random sample of women stratified by site and enrollment in WC versus TC (total response rate = 61%). MEASURES: Overall satisfaction and gender-specific satisfaction as measured by the Primary Care Satisfaction Survey for Women (PCSSW). ANALYSIS: We dichotomized the satisfaction scores (excellent versus all other), and compared excellent satisfaction in WCs versus TCs using logistic regression, controlling for demographics, health status, health care use, and location. RESULTS: Women enrolled in WCs were more likely than those in TCs to report excellent overall satisfaction (odds ratio, 1.42; 95% confidence interval, 1.00 to 2.02; P = .05). Multivariate models demonstrated that receipt of care in WCs was a significant positive predictor for all 5 satisfaction domains (i.e., getting care, privacy and comfort, communication, complete care, and follow-up care) with the gender-specific satisfaction instrument (PCSSW). CONCLUSIONS: This study is the first to consistently show higher satisfaction in WCs versus TCs despite age and race differences and comparable health status. Since these WCs show better quality in terms of satisfaction, other quality indicators should be evaluated. If WCs reduce fragmentation and improve health care delivery, the model will be applicable in VA and non-VA outpatient settings. PMID- 12648249 TI - Beliefs and expectations of women under 50 years old regarding screening mammography: a qualitative study. AB - OBJECTIVE: Because shared decision making has been recommended for screening mammography by women under age 50, we studied women's decision-making process regarding the procedure. DESIGN: Qualitative research design using in-depth semi structured interviews. PATIENTS: Sixteen white and African-American women aged 38 to 45 receiving care at a large New England medical practice. MEASUREMENTS AND MAIN RESULTS: We identified the following content areas in women's decision making process: intentions for screening, motivating factors to undergo screening, attitudes toward screening mammography, attitudes toward breast cancer, and preferences for information and shared decision making. In our sample, all women had or intended to have a screening mammogram before age 50. They were motivated by the awareness of the recommendation to begin screening at age 40, knowing others with breast cancer, and a sense of personal responsibility for their health. Participants feared breast cancer and thought the benefits of screening mammography far outweighed its risks. Women's preferences for involvement in decision making varied from wanting full responsibility for screening decisions to deferring to their medical providers. All preferred the primary care provider to be the main source of information, yet the participants stated that their own providers played a limited role in educating them about the risks and benefits of screening and the mammography procedure itself. Most of their information was derived from the media. CONCLUSIONS: The women in this study demonstrated little ambivalence in their desire for mammography screening prior to age 50. They reported minimal communication with their medical providers about the risks and benefits of screening. Better information flow regarding mammography screening is necessary. Given the lack of uncertainty among women's perceptions regarding screening mammography, shared decision making in this area may be difficult to achieve. PMID- 12648250 TI - Comparison of current guidelines for primary prevention of coronary heart disease: risk assessment and lipid-lowering therapy. AB - OBJECTIVE: In primary prevention of atherosclerotic disease, it is difficult to decide when medical treatment should be initiated. The main goal of the study was to compare different guidelines for coronary heart disease (CHD) risk assessment and initiation of lipid-lowering therapy. DESIGN: Cross-sectional evaluation. SETTING: An outpatient lipid and diabetes clinic in a university hospital. PARTICIPANTS/METHODS: Risk factor data obtained on 100 consecutive patients (58 men and 42 women) without clinical evidence of cardiovascular disease were used to compare the Framingham risk equation, the U.S. National Cholesterol Education Program (Adult Treatment Panel III) (NCEP ATP III) guidelines, the joint European Societies guidelines, the joint British guidelines, the revised Sheffield table, and the Munster Heart Study calculator (PROCAM) CHD risk assessment and lipid lowering therapy. RESULTS: Guidelines could be applied to different subsets of the cohort, ranging from 22% (PROCAM) to 95% of the cohort (revised Sheffield table). All guidelines (except PROCAM) could be applied to a total of 62 patients. Guidelines predicted > or =20% risk for developing CHD over 10 years in 53% (NCEP ATP III), 26% (European) and 32% (British), while Framingham predicted this risk level in 34%. CHD risk was estimated to be > or =3%/year in 5% according to Sheffield, while Framingham predicted this risk in 13%. Lipid lowering drug therapy is recommended in 52% by NCEP ATP III, while European, British, and Sheffield guidelines recommend this in 26%, 35%, and 5%, respectively. CONCLUSIONS: Guidelines for assessing CHD risk and lipid-lowering therapy differ greatly. Therefore, these algorithms must be used with caution. PMID- 12648252 TI - Could our pretest probabilities become evidence based? A prospective survey of hospital practice. AB - OBJECTIVE: We sought to measure the proportion of patients on our clinical service who presented with clinical problems for which research evidence was available to inform estimates of pretest probability. We also aimed to discern whether any of this evidence was of sufficient quality that we would want to use it for clinical decision making. DESIGN: Prospective, consecutive case series and literature survey. SETTING: Inpatient medical service of a university-affiliated Veterans' Affairs hospital in south Texas. PATIENTS: Patients admitted during the 3 study months for diagnostic evaluation. MEASUREMENTS: Patients' active clinical problems were identified prospectively and recorded at the time of discharge, transfer, or death. We electronically searched medline and hand-searched bibliographies to find citations that reported research evidence about the frequency of underlying diseases that cause these clinical problems. We critically appraised selected citations and ranked them on a hierarchy of evidence. RESULTS: We admitted 122 patients for diagnostic evaluation, in whom we identified 45 different principal clinical problems. For 35 of the 45 problems (78%; 95% confidence interval [95% CI], 66% to 90%), we found citations that qualified as disease probability evidence. Thus, 111 of our 122 patients (91%; 95% CI, 86% to 96%) had clinical problems for which evidence was available in the medical literature. CONCLUSIONS: During 3 months on our hospital medicine service, almost all of the patients admitted for diagnostic evaluation had clinical problems for which evidence is available to guide our estimates of pretest probability. If confirmed by others, these data suggest that clinicians' pretest probabilities could become evidence based. PMID- 12648251 TI - Clinically important changes in health-related quality of life for patients with chronic obstructive pulmonary disease: an expert consensus panel report. AB - OBJECTIVE: Without clinical input on what constitutes a significant change, health-related quality of life (HRQoL) measures are less likely to be adopted by clinicians for use in daily practice. Although standards can be determined empirically by within-person change studies based on patient self-reports, these anchor-based methods incorporate only the patients' perspectives of important HRQoL change, and do not reflect an informed clinical evaluation. The objective of this study was to establish clinically important difference standards from the physician's perspective for use of 2 HRQoL measures among patients with chronic obstructive pulmonary disease (COPD). DESIGN: We assembled a 9-person expert panel of North American physicians familiar with the use of the Chronic Respiratory Questionnaire (CRQ), a disease-specific HRQoL measure, or the generic Medical Outcomes Study Short-Form 36-Item Health Survey (SF-36, Version 2.0) among patients with COPD. RESULTS: Using 2 rounds of the Delphi process, 1 in person meeting, and an iterative improvement process for circulating and correcting the final report, the expert panel established small, moderate, and large clinically important change levels for the CRQ and SF-36. CONCLUSIONS: For this expert physician panel, levels for detecting clinically important differences on the CRQ were equal to or slightly higher than previous studies based on patient-reported differences. Clinically important differences on the SF 36, Version 2.0, were noticeably larger than previous estimates based on cross sectional differences between clinically defined patient groups. PMID- 12648253 TI - What do academic primary care physicians want in an electronic journal? AB - To determine the interest of academic general internists and family physicians in specific features of electronic journal publications, we surveyed 350 physicians, 175 randomly selected from each of 2 medical societies: the Society of General Internal Medicine, and the Society of Teachers of Family Medicine. The response rate was 70%. Most general internists and family physicians used online journals sometimes or often. Most general internists and family physicians reported moderate to high interest in having links from original articles, reviews, or editorials to listed references (77% to 89% of internists and 65% to 81% of family physicians) and electronic medical reference texts (73% to 78% of internists and 65% to 83% of family physicians). Less than 25% of both groups reported moderate to high interest in having links to initiate dialog with other readers or to communicate comments to the author or editor. General internists were more likely than were family physicians to have moderate to high interest in having links to appendices and supportive material (e.g., 66% of general internists versus 46% of family physicians for original articles; P < .05) and less likely to have moderate to high interest in links to health-related web sites (44% of general internists versus 69% of family physicians for original articles; P < .05). We conclude that academic general internists and family physicians have strong but not identical interests in specific features of electronic publication that primary care-oriented journals should consider. PMID- 12648254 TI - Rehabilitation for patients with chronic obstructive pulmonary disease: meta analysis of randomized controlled trials. AB - OBJECTIVE: To develop a meta-analysis to determine the effectiveness of rehabilitation in patients with chronic obstructive pulmonary disease (COPD). DATA SOURCES: medline, cinhal, and Cochrane Library searches for trials of rehabilitation for COPD patients. Abstracts presented at national meetings and the reference lists of pertinent articles were reviewed. STUDY SELECTION: Studies were included if: trials were randomized; patients were symptomatic with forced expiratory volume in one second (FEV1) <70% or FEV1 divided by forced vital capacity (FEV1/FVC) <70% predicted; rehabilitation group received at least 4 weeks of rehabilitation; control group received no rehabilitation; and outcome measures included exercise capacity or shortness of breath. We identified 69 trials, of which 20 trials were included in the final analysis. DATA EXTRACTION: Effect of rehabilitation was calculated as the standardized effect size (ES) using random effects estimation techniques. RESULTS: The rehabilitation groups of 20 trials (979 patients) did significantly better than control groups on walking test (ES = 0.71; 95% confidence interval [95% CI], 0.43 to 0.99). The rehabilitation groups of 12 trials (723 patients) that used the Chronic Respiratory Disease Questionnaire had less shortness of breath than did the control groups (ES = 0.62; 95% CI, 0.35 to 0.89). Trials that used respiratory muscle training only showed no significant difference between rehabilitation and control groups, whereas trials that used at least lower-extremity training showed that rehabilitation groups did significantly better than control groups on walking test and shortness of breath. Trials that included severe COPD patients showed that rehabilitation groups did significantly better than control groups only when the rehabilitation programs were 6 months or longer. Trials that included mild/moderate COPD patients showed that rehabilitation groups did significantly better than control groups with both short- and long-term rehabilitation programs. CONCLUSION: COPD patients who receive rehabilitation have a better exercise capacity and they experience less shortness of breath than patients who do not receive rehabilitation. COPD patients may benefit from rehabilitation programs that include at least lower-extremity training. Patients with mild/moderate COPD benefit from short- and long-term rehabilitation, whereas patients with severe COPD may benefit from rehabilitation programs of at least 6 months. PMID- 12648256 TI - Diagnosis: highlighting the gaps. PMID- 12648255 TI - Host-environment medicine: a primary care model for the age of genomics. AB - Rapid developments in genomic and proteomic testing promise to impact the way in which clinicians assess disease risk and drug selection in their patients. Because most diseases result from host-environment interactions, however, primary care providers will need to avoid the trap of biological determinism by examining the important role of environmental factors in their clinical assessments and interventions. This article discusses the application of host-environment concepts to recent developments in the areas of genomics and proteomics. PMID- 12648257 TI - Declining interest in primary care careers. PMID- 12648258 TI - The assessment of systematic reviews in dentistry. AB - The aim of this study was to assess the quality of systematic reviews of effectiveness of interventions in dentistry. The Database of Abstracts of Reviews of Effectiveness and the Cochrane Database of Systematic Reviews were searched to identify systematic reviews examining the effectiveness of interventions for oral, dental and craniofacial disorders and diseases. Sixty-five reviews were identified and assessed independently by two reviewers. The area most frequently evaluated within the reviews was pain relief/prevention (20/65, 31%) followed by caries and oral medicine. The quality assessment of the identified systematic reviews highlighted key areas where improvements could be made. One major weakness of the reviews was that the search strategies employed were not always adequate. Only 12 reviews (19%) demonstrated an attempt to identify all relevant studies. Other areas of weakness include the screening and quality assessment of primary studies, the pooling of data and examination of heterogeneity, and the interpretation of findings. This investigation shows that the quality of systematic reviews in dentistry could be improved. If future clinical decisions are to be based upon systematic reviews, it is imperative that the reviews address clinically relevant, focused questions and follow a 'transparent', well designed protocol. PMID- 12648259 TI - Temperament and character personality dimensions in patients with dental anxiety. AB - The aim of the present study was to investigate character and temperament dimensions of personality in six men and 31 women (aged 20-57 yr) with severe dental anxiety, and to evaluate whether these dimensions were associated with the level of dental anxiety. The Dental Anxiety Scale (DAS) and the Temperament and Character Inventory (TCI) were used. High ratings in novelty seeking and female gender predicted high DAS scores. Compared with controls, the patients scored significantly higher on the temperament dimension, novelty seeking. For character dimensions, the patients scored lower on cooperativeness and higher on self transcendence than controls. Our results indicated that patients with dental anxiety are neurotic extravert (i.e. novelty seekers who experience brief dissociative periods and magical thinking). Furthermore, the combination of the inherited temperament dimension novelty seeking and the social learned character dimension cooperativeness and self-transcendence seem to form a vulnerable personality to develop dental anxiety. PMID- 12648260 TI - The importance of dental beliefs for the outcome of dental-fear treatment. AB - This study investigated the importance of dental beliefs and the predictive value of the Dental Belief Survey (DBS) in dental-fear treatment. The sample comprised 117 adult patients seeking treatment at a dental-fear clinic. Pretreatment data were collected during a screening procedure, including two visits to the dentist. Outcome measurements were completed after treatment. The dentist rated successful/unsuccessful treatment outcome. Patients unsuccessful in treatment (n = 48) reported more initial negative dental beliefs, while patients successful in treatment (n = 69) showed a larger decrease in negative beliefs between the first and second visit to the dentist. However, these differences were small. There was a significant difference between the groups at visit two. Thus, patients unsuccessful in treatment reported more negative beliefs about how dentists communicate. Regression analyses showed that improved dental beliefs during the first two visits to the dentist predicted dental-fear reduction, while longer avoidance time, female gender, low engagement in treatment, and depressed mood increased the risk of unsuccessful treatment outcome. Our results suggest that the DBS provides valuable information, and that patients' subjective perceptions about how dentists communicate are important for treatment outcome. However, initial dental beliefs were not found to predict clinical treatment outcome. PMID- 12648261 TI - Impact of caries experience in the deciduous molars on the emergence of the successors. AB - The purpose of this study was to evaluate the effect of caries experience in a primary molar (decayed and/or restored vs. extracted) on the timing of emergence of its successor. Data were obtained from a longitudinal epidemiological study in a sample of 4468 Flemish children. Survival analyses with log-logistic distribution were performed to calculate median emergence ages and 95% confidence intervals (CI); caries experience was added as a covariate. The results indicate that the emergence of the maxillary and mandibular premolars was accelerated by 2 8 months when its predecessor had been decayed and or restored but had not been extracted. Premature loss of maxillary primary molars resulted in a significant acceleration of the emergence of the premolars; this was not observed in the mandible. In conclusion, when considering permanent tooth emergence ages, caries experience in the primary dentition should be taken into account. PMID- 12648262 TI - The effect of retention on orthodontic relapse after the use of small continuous or discontinuous forces. An experimental study in beagle dogs. AB - Relapse is a major concern in orthodontics for which avoidance retention is the general procedure. However, the effect of retention on relapse after active tooth movement with different force regimes has never been studied in a standardized experimental setting. Mandibular third premolars were extracted in 19 young adult beagle dogs. Three months later, the second premolars were bodily moved distally with forces of 10 cN or 25 cN. The forces were applied for 24 h d-1 or for 16 h d 1. After 4 months, relapse was allowed in half of the animals, while in the others relapse was preceded by retention for 90 d. Statistical analyses were performed on the relation between force regime, active tooth movement, retention, and relapse. Force magnitude had no effect on relapse, while continuous forces resulted in a longer-lasting and more pronounced relapse than did discontinuous forces. A significant positive correlation was found between the amount of active tooth movement and both the rate and the total amount of relapse, but not between the amount of active tooth movement and the duration of the relapse. Retention had no effect on the duration of the relapse, but it strongly decreased its total amount. Finally, the effect of retention on the amount of relapse was strongly correlated with the amount of active tooth movement. PMID- 12648263 TI - Expression profile of matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs in mature human odontoblasts and pulp tissue. AB - Previous studies have demonstrated that (at least) matrix metalloproteinase (MMP) 2, -8, -9, -14 and -20 are expressed by human odontoblasts. Here, we analysed the expression of 19 MMPs and their specific tissue inhibitors (TIMP)-1, -2 and -3) 1, -2 and -3 in mature human odontoblasts and pulp tissue. Since MMP-20 is almost exclusively expressed by the dentin-pulp complex cells, we further analysed the effect of transforming growth factor (TGF)-beta1 and bone morphogenetic protein (BMPs)-2 on its expression. Matrix metalloproteinase-9 served as a positive control for growth factor responsiveness. It was found that MMP-1, -2, -9, -10, 11, -13, -14, -15, -16, -17, -19, -20 and -23, in addition to TIMP-1, -2 and -3 were expressed by both odontoblasts and pulp tissue. Neither MMP-3 nor MMP-12 were expressed in odontoblasts or pulp tissue, and MMP-7, -8, -24 and -25 were expressed only in the odontoblasts; MMP-2, -10, -11, -14 and -20 were expressed more abundantly by odontoblasts, whereas pulp tissue expressed more MMP-13 and MMP-17. Transforming growth factor-beta1 (1 ng ml(-1)) and BMP-2 (100 ng ml(-1)) did not markedly affect MMP-20 mRNA expression. In contrast, TGF-beta1 alone and with BMP-2 significantly upregulated MMP-9 mRNA by 2.4-fold and by 2.6-fold, respectively, in odontoblasts, while in pulp tissue no effects could be detected. The wide-scale expression of MMPs and TIMPs by mature human odontoblasts and pulp tissue suggests that they may participate in dentin matrix organization prior to mineralization, and that growth factors may further control dentin matrix modeling by differentially regulating individual MMPs. PMID- 12648264 TI - Identification, distribution and expression of osteoadherin during tooth formation. AB - Osteoadherin (OSAD) is a keratan sulfate-containing proteoglycan, belonging to the small leucine-rich proteoglycan (SLRP) family. In bone OSAD has been localized in primary spongiosa within the bovine fetal rib growth plate. Moreover, in situ hybridization has shown expression of OSAD in osteoblasts close to the cartilage and bone border in the growth plate of rat femur. mRNA expression has also detected OSAD in mature osteoblasts on the surface of bone trabeculae. We have identified OSAD in extracts of bovine dentin, and the identity was verified by N-terminal sequencing. Western blot analysis detected two bands in bovine bone and dentin at 85 kDa and 60 kDa. Northern blot analysis of RNA samples from 5-d-old-rat tooth and femur showed a 1.9-kb transcript for OSAD in both tissues. OSAD was located to the mineralized dentin matrix, cementum and surrounding alveolar bone by immunohistochemistry, and in situ hybridization showed OSAD to be highly expressed during early crown formation in the entire odontoblast cell layer, in the area of Hertwig's epithelial root sheath, in the cells of the newly formed mantle dentin, and in the odontoblasts at the fissures. Ultrastructural studies indicated that OSAD might be associated with collagen fibrils. Thus, OSAD may play an important role during tooth development and biomineralization of dentin. PMID- 12648265 TI - Influence of the endothelium on the vasorelaxant response to acetylcholine and vasoactive intestinal polypeptide in the isolated rabbit facial artery. AB - The aim was to examine the influence of the endothelium on acetylcholine (ACh) and vasoactive intestinal polypeptide (VIP) functional responses in the isolated glandular branch of rabbit facial artery precontracted with phenylephrine as well as the potential contribution of nitric oxide (NO) and prostanoids in the ACh- and VIP-induced effects. Acetylcholine caused endothelium-dependent and VIP endothelium-independent relaxations of facial artery. The effect of ACh was partly inhibited by NG-monomethyl-l-arginine (l-NMMA, a non-selective NO synthase inhibitor) or by indomethacin (a cyclooxygenase inhibitor) while being completely blocked after concomitant addition of l-NMMA and indomethacin. The relaxation of the facial artery caused by ACh was unaffected by 65 mm KCl. The VIP-induced vasodilation was potentiated by forskolin (an adenylate cyclase stimulator) and partly reduced by l-NMMA or S-methyl-l-thiocitrulline (l-SMTC, a neuronal NO synthase inhibitor), whereas it was unaffected by indomethacin. These results suggest that ACh effects on the rabbit facial artery are mediated through release of endothelium-derived NO and cyclooxygenase products, while the effect of VIP is most probably mediated by an increase of cyclic adenosine 3',5'-monophosphate (cAMP) in vascular smooth muscles and by VIP-induced release of NO from perivascular nerve fibers. PMID- 12648266 TI - Comparison of antiadhesive and antibacterial effects of antiseptics on Streptococcus sanguinis. AB - Three antiseptic (chlorhexidine, Olaflur, Octenisept) and one putative antiadhesive (chitosan) agent were investigated for their effect on viable planktonic and attached Streptococcus sanguinis cells. The bacterial pretreatment with each chemotherapeutic was performed in two steps: (i) After the exposure of planktonic streptococci to the antiseptics, the cells were suspended in human sterile saliva and allowed to attach to human enamel for 60 min; (ii) After 60 min in the flow chamber system, initially attached streptococci were treated with these agents. The microbial viability was monitored by the percentage of vital streptococci determined by fluorescence microscopy and cell reproduction. In comparison with the negative control NaCl, the non-bactericidal chitosan derivative showed distinctive antiadhesive properties. For both treatment procedures, the efficacy of the antiseptics in reducing the viability of planktonic and attached streptococci was Octenisept > Olaflur > chlorhexidine > saline > chitosan. Further studies appear warranted to develop new antiplaque/antibiofilm strategies involving highly efficient bactericidals with antiadhesive formulations. PMID- 12648267 TI - Abfraction lesion formation in maxillary incisors, canines and premolars: a finite element study. AB - Abfraction lesions are angular, wedge-shaped defects found at the cervical region of teeth and are caused by mechanical overloading initiated by cuspal flexure. Clinically, these lesions are more prevalent on the labial aspect of maxillary incisors. The aim of this study was to provide a biomechanical explanation for this clinical variation. Two-dimensional plane strain finite element models of an maxillary incisor, canine and first premolar were developed and the cervical stress profiles were examined along a horizontal plane 1.1 mm above the amelo cemental junction. The local X (horizontal) stress on the labial/buccal side was 176.4 MPa for the incisor, 57.8 MPa for the premolar, and 3.4 MPa for the canine. Similarly, the maximum labial/buccal principal stress was 181.4 MPa for the incisor, 25.2 MPa for the premolar, and 66.8 MPa for the canine. The labial/buccal stress profile in the cervical region of an maxillary incisor was always greater than that found in an maxillary canine or premolar tooth. These findings provide a biomechanical explanation for the clinical variation seen in the prevalence of cervical abfraction lesions. PMID- 12648268 TI - Marginal deterioration as a predictor of failure of a posterior composite. AB - The purpose of the present study was to undertake a retrospective analysis of the findings of a multicentre clinical trial to test the hypothesis that marginal deterioration and cavomarginal discoloration may be a predictor of the failure of posterior composite restorations. Longitudinal 5-yr data from the multicentre trial of Occlusin were analysed (modified USPHS criteria data were available in respect of 1018 of the 1242 restorations included in the study). The 5-yr failure rate was calculated using three different methods. The various failure rates (maximum 18.8%) and the data pertaining to marginal deterioration and cavomarginal discoloration were then compared and contrasted by means of chi2 test. The analysis revealed that restorations with marginal deterioration at 3 yr were 5.3 times more likely to have failed by 5 yr than restorations with A (Alfa) rated marginal adaptation at 3 yr. Similarly restorations with cavomarginal discoloration at 3 yr were found to be 3.8 times more likely to have failed at 5 yr than restorations with no cavomarginal discoloration at 3 yr. Moreover, restorations with both marginal deterioration and cavomarginal discoloration at 3 yr failed 8.7 times more frequently than restorations with sound margin at 3 yr. It is concluded that clinical investigations of present-day materials for posterior composite should seek to determine if marginal deterioration and cavomarginal discoloration is an important predictor of the failure of posterior composites, especially when marginal deterioration and cavomarginal discoloration occur simultaneously. PMID- 12648269 TI - Clinical and SEM evaluation of all-ceramic chair-side CAD/CAM-generated partial crowns. AB - The effect of hardware and software on the quality of Cerec all-ceramic partial crowns was investigated in this cross-sectional study. Partial crowns (n = 818) had been adhesively placed in 496 patients between 1993 and 1997 using Cerec 1 and Cerec 2 units (groups 1 and 2) as well as Cerec 2 with wall-spacing software (group 3). From each group, 25 randomly selected partial crowns were evaluated using modified United States Public Health Service (USPHS) criteria. Of these, 12 were randomly selected in each group, replicas taken and examined in a scanning electron microscope for marginal interfacial width and for continuous margin adaptation. Interfacial width of group 1 (308 +/- 95 micro m) was significantly larger than those of groups 2 (243 +/- 48 micro m) and 3 (207 +/- 63 micro m). Continuous margin adaptation at the tooth-luting composite and luting composite restoration interfaces showed only minor differences in groups 1 (94.5 +/- 8% and 95.5 +/- 2%), 2 (98.1 +/- 1% and 97.5 +/- 1.4%) and 3 (96.8 +/- 3% and 96.8 +/- 2%). Pooled clinical rating was excellent or good at 97% for all groups, indicating acceptable restoration quality except for one breakage in group 1. PMID- 12648270 TI - Adhesive bonding of titanium-aluminum-niobium alloy with nine surface preparations and three self-curing resins. AB - The purpose of the current study was to evaluate the adhesive performance of metal conditioners when used for bonding between auto-polymerizing methacrylic resins and a titanium alloy. Disk specimens were cast from a titanium-aluminum niobium (Ti-6Al-7Nb) alloy, air-abraded with alumina, and bonded with 24 combinations of eight metal conditioners (Acryl Bond, ACB; All-Bond 2 Primer B, ABB; Alloy Primer, ALP; Cesead II Opaque Primer, COP; Metafast Bonding Liner, MBL; Metal Primer II, MPII; MR Bond, MRB; Super-Bond liquid, SBL) and three autopolymerizing methacrylic resins (Repairsin, RE; Super-Bond C & B, SB; Tokuso Rebase; TR). Unprimed specimens were used as controls. Shear bond strengths were determined both before and after thermocycling (4-60 degrees C, 20, 000 cycles). The ALP-SB group recorded the greatest post-thermocycling bond strength (21.8 MPa) followed by the COP-SB group (17.8 MPa) and the MPII-SB group. The post thermocycling bond strengths of the unprimed-SB group and the ALP-RE group were statistically comparable. No significant differences were found among the nine TR resin groups, and these groups showed the lowest bond strength. In conclusion, the use of one of the three conditioners (ALP, COP, and MPII) in combination with the SB resin is recommended for bonding the Ti-6Al-7Nb alloy. PMID- 12648271 TI - Localization of the neuropeptide galanin in nerve fibers and epithelial keratinocytes of the rat molar gingiva. AB - Knowledge of the histochemical substrates of cellular and neurovascular connections in the gingiva is essential in order to understand the initial mechanisms of inflammation in the periodontium. Since the localization of the neuroendocrine peptide galanin in the gingiva is still unclear, we used immunohistochemical, in situ hybridization and immunoblot techniques to assess the localization of galanin in the gingiva of rat molars. Galanin-immunoreactive nerve fibers were located around blood vessels in the lamina propria, beneath the epithelium, in the epithelial-proprial junction and in the basal layer of the epithelium. Galanin was highly expressed in the suprabasal keratinocytes of the gingival epithelium. The localization of galanin in gingival nerve fibers and the expression of galanin in keratinocytes of the gingival epithelium indicate that galanin may be a possible regulator of different cellular functions in the gingiva. PMID- 12648272 TI - Computer-automated caries detection tools: a big payoff is possible when user judgment is combined with the tool. PMID- 12648275 TI - Frequencies of TAP1 and TAP2 gene polymorphisms in the Anatolian population. AB - Anatolia has a long and complex record of immigration from various regions. Here, we have used TAP1 and TAP2 gene polymorphisms as genetic markers to study the relationship between the Anatolian population and other populations. A neighbour joining tree was constructed indicating the relatedness of European populations and the Anatolian population with respect to TAP1 and TAP2 allele frequencies. PMID- 12648276 TI - Identification of novel single nucleotide polymorphisms within the NOTCH4 gene and determination of association with MHC alleles. AB - Mapping of disease susceptibility loci within the MHC has been partly hampered by the high degree of polymorphism of the HLA genes and the high level of linkage disequilibrium (LD) between markers within the MHC region. It is therefore important to identify new markers and determine the level of LD between HLA alleles and non-HLA genes. The NOTCH4 gene lies at the centromeric end of the MHC class III region, approximately 335 kb telomeric of the DRB1 locus. The encoded protein is an oncogene that is important in regulating vascular development and remodelling. A recent report has linked polymorphisms within NOTCH4 with risk of developing schizophrenia. We have investigated if coding polymorphisms exist within this gene and have identified three single nucleotide polymorphisms; a synonomous T to C transition at +1297 (HGBASE accession number SNP000064386), a synonomous A to G transition at +3061 (SNP000064387) and an A to G transition at +3063 which results in a replacement of glycine with aspartic acid at amino acid 279 (SNP000064388). The allele frequencies of +1297T, +3061A and +3063G were 0.65, 0.66 and 0.66, respectively. Linkage disequilibrium was detected both between these markers and with MHC alleles. These findings can be used in the fine mapping of disease susceptibility alleles within the MHC. PMID- 12648278 TI - HLA-DPB1*0401 is associated with dominant protection against type 1 diabetes in the general Saudi population and in subjects with a high-risk DR/DQ haplotype. AB - The human leukocyte antigen (HLA) class II DQB1*0201/0202-DRB1*04 genotype has been identified as predisposing to type 1 diabetes [insulin-dependent diabetes mellitus (IDDM)] in the Saudi Arabian population (P = 0.0002; odds ratio = 0.67; 95% confidence interval = 0.009-0.381). In this study, we searched for a factor at the DPB1 locus by analysing DPB1 polymorphism using sequence-based typing in 86 Saudi IDDM patients and control subjects, all carrying the HLA-DRB1*04/DQB1*02 haplotype or the known susceptibility allele DQB1*0201/0202. Significant protection was conferred by DPB1*0401, which was observed in 17 of 50 control subjects (55%) and 2 of 36 IDDM patients (5%) with the DQB1*0201/0202 allele (P = 0.0012; odds ratio = 8.75; confidence interval = 1.72-59.70). Our data showing a high frequency of the DPB1*0401 allele even in the presence of the predisposing DQB1*02 allele in healthy subjects may indicate a protective effect of this combination of HLA alleles against type 1 diabetes. This finding supports the hypothesis that protective HLA class II genes can override the risk conferred by HLA-DQ susceptibility alleles. Further studies using larger cohorts of control subjects and patients should be undertaken to confirm this observation. PMID- 12648277 TI - Molecular cloning of feline tumour necrosis factor receptor type I (TNFR I) and expression of TNFR I and TNFR II in lymphoid cells in cats. AB - Tumour necrosis factor (TNF)-alpha is a pro-inflammatory cytokine produced by many types of cells. It has been shown that two distinct TNF receptors (TNFRs), TNFR type I (TNFR I) and TNFR type II (TNFR II), have different functions in signal transduction, which is possibly associated with the development of a variety of diseases. In this study, we isolated a feline TNFR I cDNA clone and analysed the expression of TNFR I and TNFR II mRNA in feline lymphoid cells. The deduced amino acid sequence of feline TNFRI cDNA showed 75.8, 62.5 60.9 and 72.1% similarity with those of its human, mouse, rat, and pig counterparts, respectively. The feline TNFR I cDNA was shown to encode extracellular, transmembrane and intracellular domains fundamentally conserved in the homologues of other species. Expression of TNFR I and TNFR II mRNAs was shown to be up regulated in feline peripheral blood mononuclear cells (PBMC) by stimulation with concanavalin A. Five of six feline lymphoma cell lines were shown to express both TNFR I and TNFR II mRNAs. The expression of TNFR I in PBMC was up-regulated in cats infected with feline immunodeficiency virus (FIV), whereas the expression of TNFR II in PBMC was not different between FIV-infected cats and uninfected cats. The present study indicate that expression of TNFR I and TNFR II may be associated with disease progression, especially in retrovirus infections in cats. PMID- 12648280 TI - Distribution of human CD30 gene promoter microsatellite alleles in healthy and human immunodeficiency virus-1 infected populations. AB - The human CD30 gene, found on chromosome 1; 1p36, contains a microsatellite of the type [(CCAT)2-12CCACTTATGCAT]n within the promoter. As the microsatellite has been shown to be both polymorphic and involved in the transcriptional regulation of this gene, it is of potential interest with respect to interindividual differences in CD30 expression. Here we describe a method for determining length variation of this repeat region and determine the frequency and distribution of alleles of the CD30 microsatellite within the Western Australian population. As soluble CD30 levels are predictive for disease associated with human immunodeficiency virus (HIV)-1 infection, we also determined allele frequencies in an HIV+ cohort. PMID- 12648279 TI - Increased frequency of the mannose-binding lectin LX haplotype in Chinese systemic lupus erythematosus patients. AB - Mannose-binding lectin (MBL) is an important complement-activating protein of the human immune system. As a result of one of three structural gene mutations in exon 1 (variants B, C and D) and/or the presence of a low-efficiency promoter polymorphism, MBL deficiency may be associated with increased susceptibility to infectious diseases and to autoimmune disorders, including systemic lupus erythematosus (SLE). Using a combined approach of heteroduplex generator and polymerase chain reaction, a systematic search for mutations in exon 1 and the promoter region of the MBL gene was performed in a Chinese study population comprising 41 SLE patients and 111 healthy controls. Two alleles, a wild-type allele A and a variant allele B (a previously reported mutation of GGC to GAC at codon 54), were identified in MBL exon 1. The frequency of the B allele (0.15) was higher in the SLE patients than in the healthy controls (0.09), but the difference did not attain statistical significance (P > 0.05). However, for two polymorphisms at positions -550 and -221 in the promoter region, the frequency of the low-MBL-producing haplotype (LX) in the patients (0.2073) was significantly higher than that in the controls (0.0855) (P = 0.003, relative risk = 2.79). Our results suggest that the LX haplotype represents a strong risk factor among Chinese SLE patients. Although of lesser importance, the MBL B allele also may be a risk component in the developing process of SLE in Chinese patients. PMID- 12648281 TI - Immunogenetics of the rare HLA-B allele B*4408. AB - The nucleotide sequence of exons 2 and 3 of B*4408 has been confirmed. This allele has now been found in five, apparently unrelated, north-western European Caucasoid subjects, residing in Wales, on HLA haplotypes bearing: A*0201, Cw*0501 with DRB1*0401, DQA1*03, DQB1*0301 in four instances and DRB1*0701, DQA1*0201, DQB1*0202 in one instance. The unique serological reactivity of 'HLA-B44BO' was established using up to nine B*4408 subjects and was easily identified using a variety of B44 and B62 antisera. The phenotype and gene frequencies of B*4408, in a sample of 40 473 subjects residing in Wales, were 0.01235% and 0.00006, respectively. PMID- 12648282 TI - Polymorphisms of the TAP1 and TAP2 genes in human alveolar echinococcosis. AB - We postulated that TAP genes may influence the susceptibility of some individuals to Echinococcus multilocularis infection. Six coding region variants (codons 333 and 637 in TAP1, and 379, 565, 651 and 665 in TAP2) were typed in 94 patients and 100 controls. Thr/Thr homozygosity at TAP2/665 was more prevalent in patients than in controls [64% vs. 45%, respectively; odds ratio (OR) = 2.1 (95% confidence interval (CI) 1.1; 2.7)] and Thr/Ala heterozygozity was less prevalent (32% vs. 50%, respectively) (P = 0.014). Of the 38 patients with progressive lesions, 76% were Thr/Thr, as compared with 55% of patients without progressive lesions and 45% of controls (P = 0.058 and 0.02, respectively), independent of HLA status. To determine whether this association is functionally relevant, functional analyses and/or confirmation in distinct populations of patients with alveolar echinococcosis would be required. PMID- 12648283 TI - Relative quantification of HLA-DRA1 and -DQA1 expression by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). AB - Polymorphism in the upstream regulatory region (URR) of the MHC class II DQA1 gene defines 10 different alleles named QAP (DQA1 promoter). In vitro studies have suggested that allelic polymorphism in the HLA-DQA promoter region may result in differences in HLA-DQA1 gene expression. In the present study, we used real-time reverse transcriptase-polymerase chain reaction (RT-PCR) to quantify differences in HLA-DQA1 gene expression. After the isolation of total mRNA, reverse transcription into cDNA was carried out using random hexamer priming and moloney murine leukaemia virus (MMLV) reverse transcriptase. Quantification of DQA1 mRNA species using a set of six group-specific primer pairs for the detection of HLA-DQA1*01, *02, *03, *04, *05 and *06 was carried out on an ABI PRISM GeneAmp 7700 Sequence Detection System (Perkin Elmer, Foster City, CA) with real-time detection and quantification taking advantage of the fluorescence TaqMan technology (Perkin Elmer, Foster City, CA). Normalization of cDNA templates was achieved by glyceraldehyde-3-phosphate dehydrogenase (GAPDH) quantification. In addition, the total amount of mRNA produced by HLA-DQA1 and HLA-DRA1 expression was quantified for comparison. Subsequently, this approach was validated using Raji and HUT-78 cell lines and tested with peripheral mononuclear cells (PBMC) of 45 samples taken from healthy volunteers. The sensitivity was determined with > or = 10(2) copies. Comparison of the allele specific DQA1 expression with the total expression of DQA1 and DRA1 mRNA indicated that DQA1*04 expression was increased compared with the expression of other alleles of the DQA1 gene. Thus, allele-specific quantification of DQA1 gene products could be achieved by real-time RT-PCR suitable for the analysis of differential expression of DQA1 mRNAs in homozygote and heterozygote combinations. PMID- 12648284 TI - Allele frequencies for an interferon-gamma microsatellite in a population of Brazilian leprosy patients. AB - A group of Brazilian leprosy patients and controls were genotyped for a CA-repeat microsatellite polymorphism within the interferon (IFN)-gamma gene. A significantly higher frequency of alleles 5-7 was observed in this patient population, indicating that IFN-gamma gene polymorphism may contribute to the course of leprosy post-infection. PMID- 12648285 TI - HLA-B*15 subtypes in the population of north-eastern Thailand. AB - The HLA-B*15 group is the most polymorphic HLA-B allele and so has several subtypes. These subtypes have not been defined in the population of north-eastern Thailand (NET). In a previous study, using polymerase chain reaction-sequence specific primers (PCR-SSP), subtypes were categorized into four groups, namely: group I: HLA-B*15 (01, 04-07, 12, 14, 19, 20, 24, 25, 26N, 27, 32, 33, 34 and 35); group II: HLA-B*15 (02, 08, 11, 15, 28 and 30); group III: HLA-B*1503/4802; group IV: HLA-B*1521. Groups I and II occurred frequently (allele frequency = 8.0 and 2.5%), and thus we optimized the polymerase chain reaction-single-stranded conformation polymorphism (PCR-SSCP) method to identify HLA-B*15 subtypes of groups I and II. Eighty samples of DNA carrying HLA-B*15 from 300 healthy unrelated individuals were tested. B*1502 (52.5%) and B*1525 (13.8%) were the most common subtypes found in NET. They also showed strong linkage disequilibrium with HLA-Cw and heterogeneity of HLA-A, DR, DQ haplotypes. Although limited conclusions can be drawn from this study because of the small number of DNA references used, the baseline data will be useful in the selection of common HLA B*15 alleles when subtyping for unrelated donor transplantations. PMID- 12648286 TI - Expression of low-frequency Ala108Pro substitution in the platelet glycoprotein Ibbeta gene. AB - We determined the gene frequency of the glycoprotein (GP) Ibbeta Ala108Pro substitution. The Pro108 allele was not found in 208 healthy Japanese and 200 healthy Caucasians. In vitro expression studies showed surface expression of the GPIbbeta Pro108 variant, suggesting the possibility of the involvement of the substitution as an alloantigen. PMID- 12648287 TI - Levels of soluble HLA-G in amniotic fluid are related to the sex of the offspring. PMID- 12648288 TI - Nomenclature for factors of the HLA system, update November 2002. PMID- 12648289 TI - Nomenclature for factors of the HLA system, update December 2002. PMID- 12648291 TI - Contraception and society. AB - When an idea meets the exigencies of an epoch, it becomes stronger than any form of political power and it becomes the common property of humankind. Such an idea was the development of contraceptives. In retrospect, the invention of contraceptives was as fundamental for the evolution of humankind as the invention of the wheel; today more than 550 million couples are using contraceptive methods. The large-scale use of contraceptives triggered the most powerful social revolutions of a century in reproductive health and gender equity, and substantially contributed to an unparalleled demographic change, characterized by a rapid aging of populations. One of the important reasons for population aging is a significant decline in fertility rates, resulting in gradually changing population structures with fewer and fewer children and more and more elderly persons. The causes underlying these demographic changes are complex and manifold; they reflect major societal changes of historical dimensions. Many of our institutions cater increasingly for a population structure that no longer exists. There is therefore an increasing need for institutional reforms in social security, health care, housing and education. In addition, several surveys conducted in the developed world have indicated an erosion of confidence in our basic institutions, e.g. courts and justice, the Church and Parliament. Whereas modem sociologists are concerned about an increase in crime, decrease in trust and depleted social capital, one can also observe an accelerated perception of our global destiny and a re-awakening of the moral impulse with a strong demand for increased transparency in public affairs. Also, various global communities have assumed a growing importance. It can be predicted that international professional communities, such as the European Society of Contraception, will play an increasingly important future role in influencing policies in general and health policies in particular. because of their profound commitment to the improvement of the human condition by the judicious use of new scientific information. PMID- 12648292 TI - Improving compliance in oral contraception: 'the reminder card'. AB - OBJECTIVES: To assess oral contraceptive compliance and to evaluate the reminder card device as a means of improving compliance. The study also aimed to evaluate ease of use of the card and women's attitudes to it after several months of use. METHODS: In France, 975 women requesting oral contraception (whether repeat prescription or first-time use) were asked retrospectively about missed pills during the previous 3 or 6 months. The decision of whether or not to provide a patient with a reminder card at inclusion was left to the gynecologist. Usefulness of and satisfaction with the reminder card were also assessed. RESULTS: Of all study participants, 60% acknowledged missing at least one pill since starting oral contraception. Compliance appeared to be significantly better in card users than non-users (p < 0.001). More than 80% of card users stated that the device had prevented them from forgetting to take the pill on at least one occasion. Forty-six per cent of prevented missed pills occurred during the first week of the cycle. No pregnancies were recorded. Among the card users, 98% found the card easy to use and 97% were satisfied with the card. CONCLUSIONS: Use of the reminder card significantly improved compliance. Users found the reminder card acceptable and easy to use. This type of reminder device could potentially reduce the number of unwanted pregnancies and hence the number of requests for pregnancy termination. PMID- 12648293 TI - The differentials and determinants of perinatal mortality in rural Bangladesh. AB - OBJECTIVE: In Bangladesh, the perinatal mortality is very high. This study examined the differentials and determinants of perinatal mortality in rural Bangladesh. METHODS: The study was based on the prospective data on maternal morbidity collected by the Bangladesh Institute of Research for Promotion of Essential and Reproductive Health and Technologies (BIRPERHT). This prospective study was conducted during the period November 1992 to December 1993. In this study, the factors associated with perinatal deaths were examined, employing differential and multivariate analyses. RESULTS: It was found that assisted delivery caused higher risks of stillbirth and death among live births during the first week of life. It was also observed that five or more pregnancies, prior to the index pregnancy, were positively associated with perinatal death. CONCLUSION: If the delivery is assisted or there are complications in delivery, then it is likely that the incidence of perinatal mortality will increase sharply. If the newborn baby was given colostrum, then perinatal mortality decreased to a great extent. PMID- 12648294 TI - Five-year continuation rate and reasons for early removal of Norplant in Shiraz, Iran. AB - Norplant has been the focus of controversy in Iran because of its relatively high premature removal rate. This study assessed the reasons for Norplant removal and its 5-year continuation rate to clarify the determining factors on elimination of this method. Subjects were identified via a retrospective chart review of Norplant users in Shiraz, Iran who were followed for 5 years. The mean age and weight of acceptors were 23.6 +/- 4.1 years and 54.4 +/- 8.5 kg, respectively. The most common reasons for removal were menstrual disturbances and personal reasons (each 41%). The 5-year continuation rate was 45%; on average, Norplant capsules were in place for 4.1 +/- 0.09 years. Norplant has been shown to be an acceptable option in other Muslim countries, so the relatively high rate of removal in Iran could not be attributed to the Norplant itself, but is probably linked to other factors that are in need of improvement. PMID- 12648295 TI - Turkish women's satisfaction with family planning services. AB - This research was designed to identify women's satisfaction with family planning services and, secondly, to establish a standardized scale of satisfaction for use in the evaluation of women's satisfaction with family planning services. The population for this study comprised Turkish women living in Erzurum, and attending one of five primary health-care centers in Erzurum for services. The study included 406 women who were selected by random sampling from the population who used current family planning methods. Data were collected using the satisfaction with family planning services scale front 1 April to 20 June 2002. Cronbach's alpha of the scale is 0.86. Alpha coefficients ranged from 0.83 to 0.77 for seven dimensions. Factor analysis on the 37 items revealed that the seven factors that resulted were the same as the seven components of intrinsic motivation. The score mean of the scale was 132.2 +/- 26.8, and the score mean per item was 3.5. In terms of both the mean overall score and the score per item, women were reasonably satisfied with family planning services. This study provided a standardized scale of women's satisfaction with family planning services. The sample of the present study was collected from only the Eastern Anatolia region of Turkey. Utilization of such methods as an interview form that would identify factors and rate their importance while women are in health-care centers and offices would be helpful. PMID- 12648296 TI - The comparison of reproductive health data in a rural district in Turkey (1981 2001). AB - OBJECTIVES: The main objective was to compare new reproductive and family planning data with data from the 1981 survey. METHODS: All 15-49-year-old married women living in seven villages of Gemlik were interviewed and the data were compared with the 1981 data of the same villages. RESULTS: In 1981, 66.8% used a family planning method, 6.8% used an intrauterine device, 10.2% used oral contraceptives and 10.0% used condoms. In 2001, these values were 93.0%, 36.5%, 6.0% and 14.8%, respectively. In 1981, 51.5% and in 2001 35.5% used coitus interruptus. CONCLUSION: In 20 years, users of both current family planning methods and modem methods have increased; users of traditional methods have decreased. PMID- 12648297 TI - Improvement of quality of life in women using a low-dose desogestrel-containing contraceptive: results of an observational clinical evaluation. AB - OBJECTIVE: To assess the influence of an oral contraceptive (OC) containing 20 microg ethinylestradiol and 150 microg desogestrel on quality of life and subjective symptoms. METHODS: In this multicenter observational evaluation performed in Germany, 3679 first-time OC users were included. They were treated by 623 physicians. The women completed quality of life questionnaires (Q-LES-Q) at baseline and after three treatment cycles. RESULTS: The mean age of users (+/- SD) was 22.5 +/- 7.1 years, with 47% of the women aged between 15 and 20 years. Half of the women reported skin problems at inclusion and, of these, 36% reported disappearance of the problems after three treatment cycles. Most women suffering from subjective symptoms (headache, nausea, nervousness, breast tenderness and depressive moods) at baseline reported improvements (57-71%) in their symptoms after using the OC for three cycles. Adverse events were reported by 4% of users; the most frequently reported were bleeding irregularities (1.7%). The total quality-of-life score and all its 13 individual items improved significantly (p < 0.0001), with the largest improvements in sex life and mood. Almost all (94%) women were either very satisfied or satisfied with the medication. CONCLUSION: The OC containing ethinylestradiol and desogestrel significantly improved the quality of life and subjective symptoms. The OC was well tolerated by first-time users. PMID- 12648290 TI - Signaling and regulation of G protein-coupled receptors in airway smooth muscle. AB - Signaling through G protein-coupled receptors (GPCRs) mediates numerous airway smooth muscle (ASM) functions including contraction, growth, and "synthetic" functions that orchestrate airway inflammation and promote remodeling of airway architecture. In this review we provide a comprehensive overview of the GPCRs that have been identified in ASM cells, and discuss the extent to which signaling via these GPCRs has been characterized and linked to distinct ASM functions. In addition, we examine the role of GPCR signaling and its regulation in asthma and asthma treatment, and suggest an integrative model whereby an imbalance of GPCR derived signals in ASM cells contributes to the asthmatic state. PMID- 12648298 TI - Two cases of abortion and premature birth after removal of a levonorgestrel releasing intrauterine system. AB - Levonorgestrel-releasing intrauterine systems (IUSs) are frequently used for contraception, increasingly also in young women before and/or between planned pregnancy. We report on two cases of abortion (double abortion and abortion followed by premature birth) after removal of a levonorgestrel-releasing IUS. Both women (25 and 26 years old) with no previous gynecological problems had a normal pregnancy and had previously delivered healthy babies. The significance of the two cases remains unclear, unless further cases are reported. However, the authors believe that there is a need for further investigation of this issue. PMID- 12648299 TI - Change of cytosolic Ca(2+) mobility in cultured bovine corneal endothelial cells by endothelin-1. AB - The effect of endothelin-1 (ET-1) on the intracellular free Ca(2+) ([Ca(2+)](i)) in cultured bovine corneal endothelial cells was studied after loading with fura 2-AM. In Ca(2+)-containing buffer and Ca(2+)-free buffer, ET-1 induced a significant rise in [Ca(2+)](i) at concentrations from 10(-9) to 10(-7) M. In Ca(2+)-free buffer, pretreatment of the cells with ET-1 inhibited thapsigargin induced [Ca(2+)](i) increase and carbonylcyanide m-chlorophenylhydrazone (CCCP) induced Ca(2+) release by 99% and 62%, respectively. Pretreatment of the cells with thapsigargin or CCCP also inhibited ET-1-induced [Ca(2+)](i) rise by 36% and 92%, respectively. In Ca(2+)-containing buffer, the ET(A) receptor antagonist (BQ123) and ET(B) receptor antagonist (BQ788) partially inhibited ET-1-induced [Ca(2+)](i) by 92% and 98%, respectively. Nifedipine and La(3+) also inhibited ET 1-induced [Ca(2+)](i) increase by 26% and 91%, respectively. The intracellular calcium release caused by ET-1 was partially inhibited by phospholipase C inhibitor (U73122). After incubation of the cells with ET-1 in Ca(2+)-free buffer, the addition of 5 mM CaCl(2) increased Ca(2+) influx, implying that release of Ca(2+) from internal stores caused by ET-1 further induced capacitative Ca(2+) entry. These data suggest that ET-1-induced [Ca(2+)](i) rise in bovine corneal endothelial cells are mediated by ET(A) receptor, ET(B) receptor, La(3+)-sensitive Ca(2+) pump and L-type voltage-operated Ca(2+) channel, leading to Ca(2+) influx. ET-1 also increased the internal Ca(2+) release from endoplasmic reticulum and mitochondria Ca(2+) stores followed by capacitative Ca(2+) entry. ET-1-induced intracellular Ca(2+) release was modulated by phospholipase C-coupled events. PMID- 12648301 TI - Conjunctival hyperemia and the use of topical prostaglandins in glaucoma and ocular hypertension. AB - The prostaglandin derivatives are thought to lower intraocular pressure (IOP) primarily by increasing uveoscleral outflow. The ocular side effect, hyperemia, appears to occur via a secondary, unrelated mechanism. Variations in the IOP lowering effect and incidence of hyperemia associated with these drugs are a function of their different chemical structures. Among the currently approved prostaglandin derivatives, hyperemia occurs in as many as 50% of patients treated with travoprost and as few as 5% of patients treated with latanoprost. The side effect of hyperemia may be of concern to the ophthalmologist for at least 2 reasons: hyperemia may compromise the outcome of filtration surgery, and it may represent a cosmetic problem to the patient thereby leading to non-compliance. The extent to which hyperemia may contribute to patient noncompliance and the effect of administration of the prostaglandin derivatives on outcome of filtration surgery remain to be determined. Until more definitive data are available, when selecting a prostaglandin analogue for ocular hypotensive therapy, it seems prudent to choose an agent with a low incidence of hyperemia. PMID- 12648300 TI - A collaborative evaluation of the cytotoxicity of two surfactants by using the human corneal epithelial cell line and the WST-1 test. AB - This study was undertaken to investigate the use of the in vitro test WST-1, an assay of cell proliferation and viability, for a preliminary safety evaluation of topical ophthalmic preparations. The cytotoxicity of two surfactants, benzalkonium chloride (BAC) and polyoxyethylene-20-stearyl ether (Brij78, PSE) was independently investigated in four laboratories in the EU by using an immortalized human corneal epithelial (HCE) cell line. The HCE cells were exposed to BAC and PSE for 5 min, 15 min, and 1 hour, and the results of the HCE-WST-1 tests were collected and compared. After one-hour exposure, the EC(50) values in BAC-treated cells in the presence of serum ranged between 0.0650 +/- 0.0284 (mean +/- SD) mM, and those in the absence of serum 0.0296 +/- 0.0081 mM. The corresponding values for PSE were 0.0581 +/-.0300 mM and 0.0228 +/-.0063 mM. There were variations in the results between different laboratories, with coefficients of variation ranging from 31 to 121%, mean 58%. The use of one-hour exposure time is to be preferred, and the elimination of serum in the culture medium is recommended to avoid both underestimation of toxic effects and variability of the test results. PMID- 12648302 TI - A 3-month comparison of efficacy and safety of brimonidine-purite 0.15% and brimonidine 0.2% in patients with glaucoma or ocular hypertension. AB - PURPOSE: To compare the efficacy and safety of brimonidine Purite 0.15% (ALPHAGAN P) BID with brimonidine 0.2% (ALPHAGAN) BID in patients with glaucoma or ocular hypertension. METHODS: 3-month, multicenter, randomized, double-masked trial. Eligible patients were taking brimonidine 0.2% BID for at least 6 weeks prior to study entry and their intraocular pressure (IOP) was < or = 21 mm Hg. Patients were randomly assigned to receive either brimonidine Purite 0.15% BID (n = 203) or brimonidine 0.2% BID (n = 204). Scheduled visits were prestudy, baseline, and weeks 2, 6 and 12. IOP was measured at hour 0 and hour 2 to evaluate efficacy. Safety was measured by monitoring adverse events. Patient satisfaction and comfort were also evaluated at all visits. RESULTS: There was no statistically significant difference between the brimonidine 0.2% and brimonidine Purite 0.15% groups with respect to mean IOP at baseline. The IOP-lowering efficacy of brimonidine Purite 0.15% was equivalent to that of brimonidine 0.2% and both study treatments maintained IOP-lowering effects of brimonidine 0.2%. There were no significant between-group differences in mean IOP. The differences in the mean IOPs were < or = 0.26 mm Hg and the mean change from baseline IOP was < or = 0.35 mm Hg at all follow-up time points. There were no statistically significant differences between the two groups in the overall incidence of adverse events. The most commonly reported treatment-related adverse events were conjunctival hyperemia and allergic conjunctivitis: both were mild in severity. CONCLUSION: Brimonidine Purite 0.15% dosed BID provides equal IOP-lowering efficacy to brimonidine 0.2% BID. Patients previously controlled on brimonidine 0.2% can be successfully switched to brimonidine Purite 0.15%. PMID- 12648303 TI - Hydrolysis of bimatoprost (Lumigan) to its free acid by ocular tissue in vitro. AB - We determined whether bimatoprost, which has been reported to act via putative prostamide receptors, could be hydrolyzed to its free acid (17-phenyl-PGF(2 alpha)), a potent FP receptor agonist, by human ocular tissue in vitro. We developed a gas chromatography/mass spectrometric method to measure 17-phenyl PGF(2 alpha) levels at sub-picomolar levels. We then analyzed the amount of 17 phenyl-PGF(2 alpha) present after incubation of 50 microl Lumigan (0.03% bimatoprost) with eye tissue using this assay. We found that cornea, sclera, iris, and ciliary body, all rapidly hydrolyzed bimatoprost to 17-phenyl-PGF(2 alpha) with linear kinetics at a rate of 6.3, 2.0, 2.8, and 1.5 pmol mg tissue( 1) hr(-1), respectively. For cornea, sclera, and ciliary body, this linear rate of hydrolysis continued over a period of at least three hours, while iris-induced hydrolysis did not continue beyond one hour. Our findings suggest that bimatoprost can act as prodrug for FP receptor activation and questions the concept of a "prostamide receptor" agonist. PMID- 12648304 TI - 5-HT(2) receptor-mediated phosphoinositide hydrolysis in bovine ciliary epithelium. AB - The serotonin 2 (5-HT(2)) receptor antagonists, MCI-9042 (Anplag) and ketanserin, have been shown to lower intraocular pressure in rabbits (1) and humans (2). The mechanism of action of these drugs has not been determined, but it is hypothesized that 5-HT(2) receptors, and possibly alpha-adrenergic receptors, (3) may regulate in part aqueous humor production via an intracellular signal transduction pathway in the ciliary body. We therefore examined whether 5-HT(2) receptors were coupled to phosphoinositide hydrolysis in an organ culture system of isolated bovine ciliary epithelium. 5-HT stimulated [(3)H]inositol phosphates ([(3)H]InsPs) accumulation in a dose-dependent manner with a maximum increase approximately twice over the basal level. The mean EC(50) value was 1.1 microM, which was calculated from four dose-response curves. The 5-HT stimulated accumulation of [(3)H]InsPs was inhibited by spiperone (5-HT(2A/1A) and dopamine 2 (D(2)) antagonists), M-1 (a major metabolite of MCI-9042), ketanserin (5-HT(2A) antagonist), SB-206553 (5- HT(2B/2C) antagonist), and mesulergine (5-HT(2C) antagonist and D(2) agonist). It was not inhibited by chlorpromazine, which is a D(2) receptor antagonist. Accordingly, our study demonstrates that 5-HT(2) receptors are coupled to phospholipase C in bovine ciliary epithelium. PMID- 12648305 TI - Amelioration by topical bunazosin hydrochloride of the impairment in ocular blood flow caused by nitric oxide synthase inhibition in rabbits. AB - We investigated whether topical instillation of an alpha(1)-adrenergic blocker would improve an insufficient blood supply in the optic nerve head (ONH) and visual function, in rabbits. The effect of systemic NOS inhibition on visual evoked potentials (VEPs) and hemodynamics in ONH were determined. VEPs were recorded before and every 15 min during a 120-min observation period after an intravenous injection of 50 mg/kg N(G)-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase (NOS) inhibitor. Capillary blood flow in ONH was evaluated by the laser speckle method throughout the same period. Then, we investigated the effect of topical instillation of a recently developed alpha(1) adrenergic blocker, bunazosin hydrochloride (0.01%), 60 min prior to the intravenous L-NAME (50 mg/kg) on the changes by NOS inhibition. The VEP amplitudes were reduced by L NAME (10, 20, and 50 mg/kg) in a dose-dependent manner, while the VEP implicit time was unchanged, and no significant changes were detected in the electroretinogram. The reductions in ONH capillary blood flow and VEP amplitudes caused by L-NAME (50 mg/kg) were significantly suppressed by an instillation of bunazosin hydrochloride. These results indicate that blocking alpha(1)-adrenergic receptors may ameliorate the impairments in blood flow and retinal function caused by NOS inhibition. The enhancement of basal vascular tone due to deprivation of continuous NO production may be diminished by this alpha(1) adrenergic blocker. PMID- 12648306 TI - In vitro inhibition of human conjunctival mast-cell degranulation by ketotifen. AB - Ketotifen relieves the symptoms of allergic conjunctivitis through multiple mechanisms of action. One such mechanism may involve stabilization of conjunctival mast cells. Because of inter- and intra-species variation, however, this hypothesis cannot be adequately tested using mast cells from animals or other human tissues. We therefore employed human conjunctival mast cells. The mast cells were prepared using human conjunctival tissues obtained from US eye banks. Cell suspensions were sensitized with human IgE and incubated with ketotifen fumarate or control. After antigenic challenge of sensitized cells with anti-IgE, levels of histamine and tryptase, two mast-cell granule markers, were measured in the supernatant fluid. Cell viability was assessed with a Trypan Blue assay. Ketotifen at concentrations of approximately 10(-11) to 10(-4) M inhibited mast-cell histamine release by 90% or more. Similarly, ketotifen at approximately 10(-10) to 10(-4) M inhibited tryptase release by 90% or more (apart from a single anomalous reading). At all ketotifen concentrations that stabilized mast cells, cell viability was preserved. Moreover, ketotifen did not impair cell viability unless concentrations were increased above the clinically relevant range, i.e., above the order of magnitude of 10(-4) M. These data demonstrate that ketotifen can stabilize human conjunctival mast cells, without impairing cell viability. PMID- 12648307 TI - The cell cycle distribution of cultured human retinal pigmented epithelial cells under exposure of anti-proliferative drugs. AB - It has been well documented that RPE cells play an important role in the formation of proliferative vitreoretinopathy (PVR). We studied the effects of four most widely used anti-proliferative drugs, i.e., 5-fluorouracil (5-FU), daunomycin (DM), mitomycin C (MMC) and dexamethasone (DEX) in an in vitro model system (cultured human RPE). The cells treated with indicated concentrations of drugs for 48 h were harvested for DNA content analysis. In addition, a time course study with constant dosage of drugs was performed. The results revealed that, after incubation for 48 h, an increased percentage (31.1%) of S phase cells was noted with exposure to MMC (10 ng/ml). It was confirmed by 5 bromodeoxyuridine (BrdU) incorporation test that this increase does not result from higher transition rate from G1 to S phase. The cells of G2/M phase markedly increased from 13.8% to 29.7% with 10 ng/ml DM. We also demonstrated that 5-FU and MMC treatment led to cell accumulation at S phase and DM treatment resulted in cell accumulation at G2/M phase, These findings were compatible with their pharmacological mechanisms. Development of an in vitro model using cultured human RPE to study the effects of various anti-proliferative drugs on cell cycle can provide a rapid, safe and inexpensive method for selection of drugs used for management of PVR. PMID- 12648309 TI - Serotyping group B streptococci in a small community hospital: an analysis of distribution and site of isolation. AB - OBJECTIVE: To determine the prevalence and site of isolation of different serotypes of group B streptococcus (GBS) colonization or infection at a small community hospital. METHODS: GBS isolates were obtained from a small community hospital and were then serotyped as la, Ib, II, III, IV, V or nontypeable. Hospital records were reviewed for patient sex, age and pregnancy status as well as the site of GBS isolation. RESULTS: GBS serotypes Ia, III and V were most common and accounted for over 60% of the total number of isolates. Serotype Ia was most prevalent in reproductive-age females, while serotypes V and III were most prevalent in non-reproductive-age females and males, respectively. Serotype la was most frequent in both pregnant and nonpregnant females. Serotype IV was more common in this study population than in those from other locations. CONCLUSIONS: The GBS serotype distribution in this small community did not differ significantly from distributions described in larger North American centers. A GBS vaccine designed against multiple serotypes would be protective for most of this population. PMID- 12648310 TI - Accuracy of five different diagnostic techniques in mild-to-moderate pelvic inflammatory disease. AB - OBJECTIVE: To evaluate the clinical diagnosis of pelvic inflammatory disease (PID) compared with the diagnosis of PID made by laparoscopy, endometrial biopsy, transvaginal ultrasound, and cervical and endometrial cultures. STUDY DESIGN: A diagnostic performance test study was carried out by cross-sectional analysis in 61 women. A group presenting PID (n = 31) was compared with a group (n = 30) presenting another cause for non-specific lower abdominal pain (NSLAP). Diagnosis provided by an evaluated method was compared with a standard diagnosis (by surgical findings, histopathology, and microbiology). The pathologist was unaware of the visual findings and presumptive diagnoses given by other methods. RESULTS: All clinical and laboratory PID criteria showed low discrimination capacity. Adnexal tenderness showed the greatest sensitivity. Clinical diagnosis had 87% sensitivity, while laparoscopy had 81% sensitivity and 100% specificity; transvaginal ultrasound had 30% sensitivity and 67% specificity; and endometrial culture had 83% sensitivity and 26% specificity. CONCLUSIONS: Clinical criteria represent the best diagnostic method for discriminating PID. Laparoscopy showed the best specificity and is thus useful in those cases having an atypical clinical course for discarding abdominal pain when caused by another factor. The other diagnostic methods might have limited use. PMID- 12648311 TI - Role of the vaginal microbiological ecosystem and cytokine profile in the promotion of cervical dysplasia: a case-control study. AB - OBJECTIVE: To identify alterations in the cytokine profile and microbial ecosystem of the vagina in association with cervical dysplasia. METHODS: Demographics, lifestyle variables and Papanicolau (Pap) smear results of subjects presenting to the same site for gynecologic complaints, obstetric visits or colposcopy were prospectively recorded. Vaginal smear for Gram stain, aerobic and anaerobic culture, pH, and wet mount and KOH examination for Trichomonas vaginalis, Gardnerella vaginalis and yeast organisms were performed. Vaginal lavage specimens were centrifuged, and the pellets and supernatants were assayed for human papillomavirus (HPV) by polymerase chain reaction and for cytokines interleukin (IL)-1beta IL-6, IL-10 and IL-12 by enzyme-linked immunosorbent assay (ELISA) respectively. Subjects with abnormal Pap smears underwent colposcopy and biopsy as indicated. RESULTS: Of 51 patients, 32 were referred for colposcopy, 12 presented with gynecologic needs, and seven presented for obstetric visits. Median age was 24 years. Demographics did not differ significantly between the dysplasia and control groups except for a trend towards more sexual partners in the dysplasia group. Biopsies were performed in 81% (26/32) of patients presenting for colposcopy and 17 revealed cervical intraepithelial neoplasia. IL 1beta, IL-6, IL-10, and IL-12 levels were elevated in 63% (20/32), 38% (15/39), 4% (2/49), and 0% of samples respectively. Elevated vaginal lavage IL-1beta was associated with a 6.1 odds ratio (95% confidence interval 1.06-35) of cervical dysplasia. Alterations in other variables studied were not associated with cervical dysplasia. CONCLUSIONS: Elevated IL-1beta, possibly representing a complex host inflammatory response to multiple pathogens, was demonstrated in patients with cervical dysplasia. PMID- 12648312 TI - Lack of an effect of protease inhibitor use on glucose tolerance during pregnancy. AB - OBJECTIVE: We hypothesized that HIV-positive women on protease inhibitors (PIs) would be more likely to have an elevated glucola test result than those not on PIs. METHODS: We reviewed our database of all HIV-positive pregnant women seen at our hospital. Serum glucose was measured 1 hour following a 50-g glucola load, at approximately 26-28 weeks of gestation. Statistical analysis was performed using Student's t-test, Fisher's exact test, and the Mann-Whitney rank sum test. RESULTS: Forty-one HIV-infected pregnant women with glucola testing were seen between January 1, 1997 and March 1, 2000. Fourteen patients were on PIs at the time of glucola. One patient in each group had an abnormal glucola test result (glucose >/= 140 mg/dl); both had normal 3-hour glucose tolerance tests. The glucola test results were similar between the PI-exposed and unexposed, with a mean difference of 5.8 mg/dl (95% confidence interval 9.2-20.8 mg/dl). Two neonates (both exposed to PI) had hypoglycemia (glucose < 40 mg/dl). CONCLUSIONS: The use of PIs does not significantly increase the risk of an elevated glucola result, nor is the mean glucola result increased in the women on PIs. The finding of hypoglycemia in neonates exposed to PIs merits further investigation. PMID- 12648313 TI - Case-control study of vulvar vestibulitis risk associated with genital infections. AB - OBJECTIVE: To evaluate the risk of vulvar vestibulitis syndrome (VVS) associated with genital infections in a case-control study. METHODS: Diagnosed cases with VVS (n = 69) and age-frequency-matched healthy controls (n = 65) were enrolled from gynecology clinics in a university medical hospital during 1999. They were compared for potential risk factors and symptoms of disease. RESULTS: VVS cases had a significantly higher risk of physician-reported bacterial vaginosis (BV) (odds ratio, OR = 9.4), Candida albicans (OR = 5.7), pelvic inflammatory disease (PID) (OR = 11.2), trichomoniasis (OR = 20.6), and vulvar dysplasia (OR = l5.7) but no risk associated with human papillomavirus (HPV), ASCUS, cervical dysplasia, genital warts, chlamydia, genital herpes or gonorrhea. Genital symptoms reported significantly more often with VVS included vulvar burning (91 vs. 12%), dyspareunia (81 vs. 15%), vulvar itching (68 vs. 23%) and dysuria (54 vs. 19%) (p < 0.0001). CONCLUSION: A history of genital infections is associated with an increased risk of VVS. Long-term follow-up case-control studies are needed to elucidate etiologic mechanisms, methods for prevention and effective treatment. PMID- 12648314 TI - Comparison of Gram stain and Pap smear procedures in the diagnosis of bacterial vaginosis. AB - OBJECTIVE: The purpose of this study was to examine the characteristics of Gram stain versus Pap smear in diagnosis of bacterial vaginosis (BV). METHODS: One thousand and sixty women were enrolled in this study. All cases with symptoms of BV were determined by Amsel's criteria, which were accepted as the gold standard for diagnosis of BV. Pap smear and Gram stain evaluations were compared according to Amsel's criteria, without viewing the clinical results of the patients. Gram stain and Pap smear results were determined as negative or positive according to Amsel's criteria. Sensitivity, specifity and positive predictive values were calculated. RESULTS: After accepting the cases that were diagnosed as BV according to Amsel's criteria as reference cases, the sensitivity of the Gram stain method was calculated as 97% and the sensitivity of the Pap smear method as 93%. Similar specificity rates were obtained with both methods in diagnosis of BV related to the clinical results. There were no statistically significant differences in diagnosis of BV between these two groups. CONCLUSION: If Amsel's criteria are accepted as the gold standard for diagnosis of BV, Gram stain and Pap smear methods will give similar results in diagnosis. PMID- 12648315 TI - Group A streptococcal toxic shock syndrome developing in the third trimester of pregnancy. AB - BACKGROUND: Group A streptococcal (GAS) toxic shock syndrome (TSS) is an uncommon, but life-threatening infection during pregnancy and should be considered in rapid onset of shock. Most cases described in the literature have occurred in the puerperium. We report a case of GAS TSS occurring during the third trimester of pregnancy in a previously healthy woman. CASE: A 31-year-old female, who was 34 weeks pregnant, presented with fevers and a prodromal 'flu like' illness. She rapidly developed shock and multiorgan failure. Blood cultures revealed GAS bacteremia and the patient met criteria for streptococcal TSS. Despite her eventual recovery, her infant died on postpartum day 15 as a consequence of the mother's TSS. CONCLUSIONS: This case is unusual in that there were no identifiable initiating events or source of the streptococcal infection, and the TSS developed during pregnancy rather than after delivery. Early recognition of GAS infections is important given the rapid onset and high morbidity and mortality associated with these infections. This is the first reported case utilizing intravenous immunoglobulin for GAS TSS in the puerperium. PMID- 12648316 TI - Two cases of delayed diagnosis of postpartal streptococcal toxic shock syndrome. AB - BACKGROUND: Puerperal sepsis due to group A beta-hemolytic streptococcal (GAS) toxic shock syndrome is associated with very high morbidity and mortality. Luckily it is now rare, but diagnosis is not always easy. This report demonstrates the problem of recognizing this disease, and summarizes the current knowledge on the pathomechanism and management of streptococcal toxic shock syndrome. CASE: Two cases of postpartum streptococcal toxic shock syndrome due to GAS are described. In both cases the correct diagnosis was delayed for several days. The first patient was sent home with the diagnosis of German measles; the second patient was transferred to our neurological intensive care unit with the diagnosis of meningitis. Both patients were admitted to the intensive care unit in profound shock, both developed multiple organ failure, and one patient died. CONCLUSIONS: GAS may produce virulence factors that cause host tissue pathology. Besides aggressive modern intensive care treatment, early diagnosis and correct choice of anti-streptococcal antibiotics are crucial. A possible adverse effect of non-steroidal anti-inflammatory agents requires confirmation in a multicenter study. PMID- 12648317 TI - Compliance with a protocol for intrapartum antibiotic prophylaxis against neonatal group B streptococcal sepsis in women with clinical risk factors. AB - OBJECTIVE: The aim of this study was to determine the prevalence of clinical risk factors (CRF) for neonatal sepsis in laboring women and to evaluate clinician compliance with a CRF-based protocol for intrapartum antibiotic prophylaxis (IAP). METHODS: A retrospective chart audit was undertaken at a district hospital (A) and a tertiary obstetric hospital (B) in Sydney, Australia between 1996 and 1998, to determine compliance with IAP in women with defined CRF. RESULTS: Eighty five (12%) women at Hospital A and 117 (19%) at Hospital B had one or more CRF. Overall compliance rates with the IAP protocols were 65 and 50% at Hospitals A and B respectively, but varied according to maternal, obstetric and sepsis related risk factors. We postulate that differences between the hospitals were related to protocol implementation. CONCLUSIONS: Compliance with a CRF-based protocol was lower than previously reported. Improvements in protocol development, implementation and maintenance are required to enhance compliance with IAP based on CRF. PMID- 12648318 TI - Zinc supplementation in Brazilian children with acute diarrhoea. AB - Although oral rehydration therapy greatly reduces mortality from diarrhoeal diseases, it has little effect on stool frequency. However, there is mounting evidence that zinc is an effective adjunct to the treatment of diarrhoea, although few studies have examined its efficacy in Latin America. This study assessed the efficacy of zinc supplementation in children with acute diarrhoea in Brazil. The study was a double-blind, placebo-controlled, randomised, clinical trial in children <5 years of age attending emergency services in Sergipe, Brazil. Subjects received zinc or vitamin C as placebo. There was a marked reduction in the duration of the diarrhoea (1.1 vs 2.6 days) and of watery stools in the zinc-supplemented group. The efficacy of zinc was independent of the presence of viral enteropathogens in the stools. It is concluded that, similar to studies in India and Bangladesh, zinc could be an important adjunct for treating acute diarrhoea in Brazilian children. PMID- 12648319 TI - Faecal lactoferrin as a predictor of positive faecal culture in south Indian children with acute diarrhoea. AB - Faecal lactoferrin, an iron-based glycoprotein found concentrated in secondary granules of neutrophils, may serve as a surrogate marker of inflammation in the intestine. We evaluated the usefulness of faecal lactoferrin as a predictor of infection with invasive enteropathogens in 262 children with diarrhoea. Lactoferrin at a dilution of 1:50 had the highest sensitivity for detection not only of conventionally cultured invasive enteropathogens but also of all other enteropathogens. Neither individual clinical symptoms nor the identification of faecal leucocytes by microscopy significantly predicted isolation of invasive enteropathogens from the faeces of children with diarrhoea. Faecal lactoferrin is a simple test which showed promise in predicting which children with diarrhoea are likely to be infected with invasive pathogens and can be incorporated as a screening test before faecal cultures are undertaken in this population. PMID- 12648320 TI - High burden of invasive Streptococcus agalactiae disease in South African infants. AB - The epidemiology of invasive Streptococcus agalactiae (GBS) disease was evaluated in South African children. Records of 208/220 children in whom GBS was isolated between January 1997 and December 1999 were reviewed. These included 63%, 31.7% and 5.3% children with early- (EOD, <7 days of age), late- (LOD, age 7-90 days) and childhood-onset disease (COD, age >90 days), respectively. The overall burden of EOD and LOD were 2.06 and 1/1000 live births, respectively. The overall mortality was 19.8% and 13.6% for infants with EOD and LOD, respectively. Risk factors for mortality in infants with EOD and LOD included septic shock (82.1% vs 1.9%), prematurity (35.2% vs 9.6%), low birthweight (29.2% vs 11.0%) and a leucocyte count <5000/mm(3) (43.5% vs 18.6%). Eight (72.7%) of 11 children with COD had an immunosuppressive, predisposing cause for invasive bacterial disease. In infants with EOD and LOD, serotype III isolates caused 49.2% and 75.7% of disease, respectively, and, together with serotype Ia isolates, caused 78.9% and 100% of invasive disease, respectively. Invasive GBS disease is common in South African infants and current strategies aimed at reducing the burden of the disease should be reconsidered. PMID- 12648321 TI - A base-line survey on birth defects in Gansu province, West China. AB - We report a study to determine the incidence, types and rank order of congenital anomalies and related fetal and infant mortality in Gansu province in China to provide a base-line for an intervention project directed at reducing birth defects in Gansu. Forty-two communities in four counties of Gansu were randomly selected by cluster sampling, based on economic and geographical features of the province. All infants born between 1 January 2001 and 1 January 2002 (live and stillborn) were investigated in departments of gynaecology and obstetrics, in birth control centres and in facilities for women and children in county and community hospitals. The types of birth defect were classified by the diagnostic standardisation ICD-9. The overall incidence of birth defects in Gansu was 15.4/1000 births (102/6621): 6.7/1000 (44) neural tube defects, 4.7/1000 (31) low birthweight infants, 2/1000 (13) limb defects, 0.8/1000 (five) cleft lip and palate and 0.5/1000 (three) Down's syndrome. The infant mortality rate was 14.8/1000 (98), to which birth defects contributed 7.3/1000 (48). We conclude that the incidence of birth defects in Gansu is one of the highest in China, that birth defects are the leading cause of infant mortality and that neural tube defects cause most deaths. PMID- 12648322 TI - Foreign body aspiration in children: experience of 1160 cases. AB - Hospital records of 1160 children 41 degrees C and abdominal pain were significantly associated with UTI. Other features significantly associated with UTI were fever of at least 7 days duration, a peak evaluation temperature >or=38.3 degrees C and a white blood cell count >10/mm(3). Escherichia coli, Klebsiella pneumonia and Staphylococcus aureus were the only organisms isolated and they all showed high in vitro sensitivity to clavulanic acid-potentiated amoxycillin, gentamicin and ceftriaxone but were poorly sensitive to co-trimoxazole. PMID- 12648325 TI - Facial paralysis caused by tuberculosis in a 2.5-month-old infant. AB - We describe an infant who presented with a cervical mass and ear discharge that did not respond to broad-spectrum antibiotics. Tuberculous infection was diagnosed after the onset of respiratory distress. Persistent otorrhoea that does not respond to conventional antibiotics or facial paralysis in a child with a discharging ear should alert the physician to a diagnosis of tuberculosis. PMID- 12648324 TI - Low level of compliance with tuberculosis treatment in children: monitoring by urine tests. AB - Patient compliance should be ensured in an effective tuberculosis control programme. We measured patient compliance by detecting antituberculous drugs in the urine of 237 outpatients receiving one to three antituberculous drugs. Positive controls were 20 hospitalised patients, supervised to receive isoniazid (INH), rifampicin (RIF) and pyrazinamide (PZA), and negative controls were not on any drugs. Among the 237 study patients, only 67% were found to be taking the appropriate treatment and 8% had taken none. We conclude that a remarkable number of patients (33%) were non-compliant with treatment. The detection of antituberculous drugs in the urine is a quick, simple and inexpensive means of measuring adherence to treatment. Unless directly observed therapy (DOT) is adopted, we recommend routine urine testing for antituberculous drugs to identify defaulting patients. PMID- 12648326 TI - Long-term outcome of preterm infants discharged home on kangaroo care in a developing country. AB - Several hospital-based studies have shown the beneficial effect of kangaroo care on preterm infants. Long-term outcome was studied in 297 preterm infants born at Harare Hospital weighing 500-1800 g, discharged home on kangaroo care and followed up for 12 months. Of these, 79 (26.6%) died, 141 (47.5%) survived to complete follow-up and 77 (25.9%) were lost to follow-up. Of those who died, median birthweight was 1460 g, median age at hospital discharge 7 days, median weight at discharge 1400 g and median age at death 66 days. Of those who completed follow-up, median birthweight was 1575 g, median age at hospital discharge was 6 days and median weight at hospital discharge was 1500 g. Of those who were lost to follow-up, median age at loss to follow-up was 70 days, median birthweight was 1540 g, median age at hospital discharge was 5 days and median weight at hospital discharge was 1500 g. The hospital re-admission rate was 22.9% with 8.8% mortality. Maternal mortality and chronic morbidity rates were 4.7% and 7.4%, respectively. On comparing those who died with those who completed follow up, mother's age <20 years, birthweight <1500 g and maternal mortality and chronic morbidity were significant risk factors for infant mortality. Age at discharge and weight at birth and on discharge were not significantly associated with infant mortality. PMID- 12648327 TI - Body composition of children in south-western Nigeria: validation of bio electrical impedance analysis. AB - Bio-electrical impedance analysis (BIA) is a non-invasive method of estimating body composition and has the potential to be useful in clinics and for nutrition and health-related research in Africa. We sought to validate BIA for use among a Yoruba population in south-western Nigeria and to use BIA to assess the body composition of a healthy cohort of children. Total body water (TBW) was measured in 92 individuals (53 adults and 39 children) using deuterium dilution; height, weight and resistance were measured by BIA. Multiple regression analysis was used to develop prediction equations for TBW among children only or among all participants. Independent covariates tested in the regression models included the impedance index (height(2)/resistance), weight, age and gender with TBW as the dependent variable. Depending on the model used, between 97% (root MSE=0.7 kg ) and 99% (root MSE=0.7 kg) of the variance observed in TBW could be explained by the impedance index, weight and/or gender; age, however, was not significant in any model. In a separate cohort of 69 children, 5-8 years old, anthropometrics were measured and TBW was estimated using the developed equations. Body composition data are presented by gender and age group. BIA was validated for use among Nigerian children and adults and provides a potentially important tool for research. PMID- 12648328 TI - A rare syndrome in the differential diagnosis of hepatosplenomegaly and pancytopenia: report of identical twins with Griscelli disease. AB - White, identical twin boys aged 3 months were referred to our centre with persisting fever, mouth ulcers, hepatosplenomegaly, pancytopenia and failure to thrive. The parents were first cousins and there was a history of a sibling with similar manifestations who had died. The infants had silvery-grey hair and pigment clumps on the hair shafts, and skin biopsy showed accumulation of melanocytes on melanosomes. Bone marrow revealed hypercellularity and haemophagocytosis. HLH-94 chemotherapy (initial therapy with daily dexamethasone and etoposide, maintenance with dexamethasone pulses, etoposide and cyclosporin A) was started. Though partial haematological remission was achieved, one of the boys died on the 34th day following aspiration pneumonia. No pathogen could be identified. The second boy responded to therapy but had a haematological relapse and died 68 days after first being admitted. Genetic study revealed a 5 bp deletion in the RAB27A gene (510 del AAGCC in exon 5). Transient haematological remission can be achieved with chemotherapy but allogeneic bone marrow transplantation is the only curative therapy in Griscelli disease, as in other familial haemophagocytic syndromes. Identification of the mutation also provides an opportunity for prenatal diagnosis. PMID- 12648329 TI - Disseminated nocardiosis in an immunocompetent child. AB - We report a 2-month-old child with a disseminated Nocardia farcinica infection that presented with suppurative lymphatic abscess. The child did not have any predisposing factors and responded to treatment with co-trimoxazole and amikacin. This is first case report of disseminated nocardiosis caused by Nocardia farcinica in an immunocompetent child. PMID- 12648330 TI - A child with supraventricular tachycardia and convulsions. AB - We present an 11-year-old girl, referred for management of episodes of supraventricular tachycardia followed by generalised tonic-clonic convulsions. The episodes did not respond to digoxin and propranolol, but subsided after initiation of carbamazepine therapy. Seizures following supraventricular tachycardia are uncommon in older children. The differential diagnosis and pathophysiology of arrhythmia-related seizures are discussed. PMID- 12648331 TI - Femoral venous access in a general paediatric setting. PMID- 12648332 TI - ThermoSpot in the detection of neonatal hypothermia. PMID- 12648333 TI - 28-Day emergency surgical re-admission rates as a clinical indicator of performance. AB - With the introduction of clinical governance, the NHS Executive has identified 28 day emergency re-admission rates as a clinical indicator to be used to assess and compare performance between NHS trusts. We undertook a 3-month retrospective audit of patients identified from the trust computer as having been re-admitted as an emergency within 28 days of discharge from the general surgical division. We wanted to examine reasons for re-admission, possible errors in coding and any preventable factors in these patients subsequently re-admitted acutely. PMID- 12648334 TI - Consent and complications: risk disclosure varies widely between individual surgeons. AB - AIMS: To assess variations in consent practice for a single operative procedure, namely laparoscopic cholecystectomy, particularly with respect to the frequency with which potential complications are discussed with patients. METHODS: Postal questionnaire sent to general surgeons in the West Midlands and to UK members of the Association of Endoscopic Surgeons of Great Britain and Ireland. Respondents were asked to estimate how often they discussed particular complications with their patients pre-operatively. RESULTS: Conversion to open cholecystectomy was the most frequently discussed complication. Other complications were much less likely to be mentioned overall and there was disturbing variation between individual surgeons in the frequency with which certain complications were discussed, particularly the risk of bile duct injury. CONCLUSIONS: In order for consent to be informed, patients need to be aware of significant risks. Our results indicate a lack of consensus from surgeons as to which risks are significant for this operation and this is likely to be true for other procedures. PMID- 12648335 TI - Variation in general practitioner referral of patients with arterial disease. AB - AIM: To compare patterns of referral for arterial disease in two areas in the UK. METHODS: A postal questionnaire was used to survey general practitioner (GP) referral patterns for arterial disease. Questionnaires were sent to West Berkshire GPs in 1993 and 2000 and to Llantrisant GPs in 2000. RESULTS: A 70-year old man with claudication at half a mile and an 80-year-old man with claudication at half a mile or 100 m were significantly more likely to be referred (P < 0.001) in West Berkshire in 2000 compared with 1993. This referral change also applied to an 80-year-old man with an aortic aneurysm who was more likely to be referred in 2000 (P < 0.01). Patients with gradual onset of rest pain were more likely to be referred urgently or as an emergency in 2000 (P < 0.05). When comparing the two areas in 2000, significant differences emerged in the likelihood of referring patients with intermittent claudication. Scepticism continues towards the value of AAA screening in West Berkshire with only about two-thirds of GPs thinking that it was of value. On the other hand in Llantrisant, 94% of GPs thought AAA screening was valuable (P < 0.001). There was a significant decrease in the number of GPs who felt that they would refer patients directly to a regional vascular centre in West Berkshire between the two time periods (P < 0.001). There was a difference in likelihood of referral to regional centres between the two areas in 2000 (33% versus 6%, P < 0.001). CONCLUSIONS: Vascular referral patterns change with time and vary from one area to another. This has implications for planning vascular services. PMID- 12648336 TI - Femoro-femoral arterial bypass is an effective and durable treatment for symptomatic unilateral iliac artery occlusion. AB - AIMS: This study was designed to determine the effectiveness of femoro-femoral arterial bypass (FFB) operation at hospital discharge and 1 year after operation, and to determine the role of long saphenous vein (LSV) as a conduit. METHODS AND RESULTS: A retrospective review was undertaken of 161 consecutive patients (median age, 66 years; range, 44-97 years) who had femoro-femoral grafts during the 12 years from July 1987 to March 1999. The indication for operation was claudication in 66 patients and critical ischaemia in 95. A synthetic graft was used in 123 patients and LSV in 38. Six patients with LSV had a previously infected synthetic graft and 2 a previously occluded synthetic graft. In-hospital operative mortality was 13 (8.1%). One year postoperatively, known mortality was 29, fifteen patients were lost to follow-up putting 1-year mortality at 18.0 27.3%. Eight of these had LSV as the conduit. At discharge from hospital, the median improvement in the ankle brachial pressure index was 0.3 (range, 0-1.0) overall, and 0.32 for patients with LSV (range 0-1.0). Among the 117 known survivors at 1 year, secondary graft patency was 107 confirmed by Doppler or duplex (91.5%) overall, and 25 (89.3%) for LSV; 100 (85.5%) maintained symptomatic improvement, 11 (9.4%) were experiencing no benefit and 6 (5.1%) were worse, of whom 2 had undergone amputation. In patients in whom LSV was used, 22 (78.8%) remained symptomatically improved, 3 (10.7%) experienced no benefit, 3 deteriorated and one had an amputation. CONCLUSIONS: One year following FFB, at least 18.0% of patients were dead. Among possible survivors to 1-year, graft patency was at least 78.8% and at least 75.8% remained clinically improved. FFB is effective in the treatment of unilateral iliac artery occlusion. LSV is as effective as a synthetic conduit. PMID- 12648337 TI - Management of gallstone disease in the elderly. AB - AIM: To determine the outcome of management of symptomatic gallstone disease (GSD) in patients aged 80 years or more. PATIENTS AND METHODS: A retrospective review of the outcome of 79 patients admitted to 2 district general hospitals with symptomatic GSD over a 1-year period was undertaken. Patients were grouped according to method of management: non-operative, ERCP, and cholecystectomy. POSSUM scores for the ERCP and cholecystectomy groups were calculated and observed, and predicted outcome compared. RESULTS: Obstructive jaundice and biliary colic were the most common presenting symptoms. Each patient had been admitted at least once before the study period (median, 2; range, 1-3). Outcomes are detailed in Table 1. Non-operative management failed in 18 of 23 patients, with 17.4% mortality. ERCP was successful in 40 of 47 patients with 3 complications (0.24 of predicted) and no mortality. In all, 11 laparoscopic and 12 open cholecystectomies were performed with 6 complications and 1 mortality (0.95 and 0.83 of predicted, respectively): 4 complications and the only death occurring after emergency cholecystectomy. Table 1 Outcomes Management Number Mortality Morbidity Outcome Non-operative 23 4 9 5 OK, 13 re-admitted, 4 still symptomatic ERCP 47 0 3 1 PTC, 6 operated Operative 23 1 6 CONCLUSIONS: This study suggests that recurrent GSD in elderly patients managed non-operatively may have fatal outcome. Elective cholecystectomy has acceptable morbidity and mortality in this age group and there is often ample opportunity to avoid emergency surgery, but a prospective randomised study is required to improve clinical algorithms. PMID- 12648338 TI - Current practice in the use of antiplatelet agents in the peri-operative period by UK vascular surgeons. AB - BACKGROUND: There currently appears to be no firm consensus with regards to the use of antiplatelet agents during the peri-operative period in vascular surgical practice. METHODS: A nine-part questionnaire relating to peri-operative antiplatelet use was sent to 137 ordinary members of the Vascular Surgical Society of Great Britain and Ireland (VSS-GBI). RESULTS: Of the 137 questionnaires sent, 90 were returned (66%). For patients undergoing infra inguinal bypass, carotid endarterectomy and varicose vein surgery, over 90% of vascular surgeons continue antiplatelet agents peri-operatively; however, in the case of aortic aneurysm repair, this figure is lower (77%). Three of the respondents stated that they would stop clopidogrel, but not aspirin, prior to surgery because of concerns over increased operative bleeding. In patients starting routine heparin prophylaxis against thrombosis, most surgeons opted to continue antiplatelet therapy (82%), although in patients requiring therapeutic heparin treatment, opinions were almost equally split. Most vascular surgeons (93%) would to start an alternative antiplatelet agent if a patient was intolerant of aspirin for gastrointestinal reasons. CONCLUSIONS: Although the benefits of antiplatelet drugs in the long-term reduction of vascular events is established, evidence supporting their use in the peri-operative period is scarce. The general consensus of opinion from this survey suggests that most vascular surgeons do not stop antiplatelet drugs pre-operatively. PMID- 12648339 TI - The fate of gallstones: traditional practice questioned. AB - AIMS: Gallstones are traditionally handed back to patients as a souvenir of their cholecystectomy. The aim of this study was to review the potential hazards of this practice, the final fate of gallstones and the patients' perception of the risks. PATIENTS AND METHODS: A total of 86 consecutive patients undergoing cholecystectomy for cholelithiasis were surveyed prospectively. An unrestricted Medline database search was performed to gather pertinent literature. RESULTS: Of the 86 patients studied, 64% took their gallstones home, out of whom 29% actually handled the stones. In 9%, gallstones were accessible to children. Of all patients surveyed, 19% thought that the stones could be harmful. No patient received any information about the potential risks. Of the study group, 35% had discarded their stones by 6 weeks and 16% were planning to discard them. The rest were unsure or had decided to keep the stones. CONCLUSIONS: Available literature establishes the presence of viable pathogenic bacteria in gallstones, but their infective potential is unknown. These bacteria are possibly more pathogenic than similar gut flora. This study has shown that gallstones are handled by a significant number of patients and are even accessible to children. The majority of patients do not consider them harmful and no information is provided to them. In this age of increasing health awareness, we recommend that patients taking their gallstones home should be informed of the potential risks. PMID- 12648340 TI - Acute adrenal insufficiency presenting as high output ileostomy. AB - We report a patient with ulcerative colitis who, in the period following surgery, developed recurrent episodes of acute adrenal insufficiency presenting clinically as a high output ileostomy. PMID- 12648341 TI - Referral guidelines for colorectal cancer--do they work? AB - AIMS AND METHODS: Urgent referral guidelines for patients with suspected colorectal cancer were introduced in 2000. In a district general hospital, we prospectively assessed the effect of these guidelines on the number of urgent referrals received and the number found to have cancer. RESULTS: Over the first year, 180 urgent referrals were received of whom 95 (55%) fitted the guidelines. Of these 95 patients, 24 (25%) had colorectal cancer. Conversely, only 2 of the 85 patients (2%) who did not fit the guidelines had colorectal cancer. During the same time period, a total of 145 new cancers were identified within the district of which 119 (82%) were in patients who had not been urgently referred to out patients as suspected colorectal cancer. DISCUSSION: The guidelines are effective in that patients who fit them have a significant chance of having colorectal cancer. However, the majority of cancers are identified outside the new system. Efforts to reduce delays in diagnosis need to recognise that many patients do not have features which fit published referral criteria. Improved support for general practitioners and better access to specialist services are required to reduce delays in diagnosis. PMID- 12648342 TI - Postoperative stay following colorectal surgery: a study of factors associated with prolonged hospital stay. AB - AIM: To study the factors that contribute to postoperative stay following colorectal surgery. DESIGN: A prospective observational study. SETTING: Three colorectal surgical units - a teaching hospital, a large district general hospital and a district general hospital. PARTICIPANTS: 350 patients undergoing colorectal surgery. MAIN OUTCOME MEASURES: 28 pre-, peri- and postoperative patient- and treatment-related factors. RESULTS: Stepwise regression analysis suggests that the factors that significantly lengthen postoperative stay include a low albumin on admission, stoma formation, operative blood loss, urinary and respiratory complications, wound infections, postoperative ventilation and social delay at the time of discharge. The postoperative stay was not affected by patient age or by the seniority of the surgical team. CONCLUSIONS: Factors have been identified that determine the postoperative length of stay. These data may allow better planning and treatment of patients undergoing colorectal surgery. PMID- 12648343 TI - Improving the quality of patient care: patient satisfaction with a nurse-led fracture clinic service. AB - A prospective study was conducted to assess nurse-led follow-up of patients with fracture. PMID- 12648344 TI - Factors delaying surgical treatment of hip fractures in elderly patients. AB - BACKGROUND: Published guidelines recommend early surgical treatment of hip fractures in elderly patients. Understanding the factors that delay surgical intervention is essential in order to introduce changes that will facilitate early treatment. AIM: To determine the factors delaying surgical treatment of hip fractures in elderly patients for more than 24 h. PATIENTS AND METHODS: Assessment of 163 consecutive patients undergoing surgery for hip fractures at the trauma unit of Manchester Royal Infirmary. RESULTS: Only 72/163 (44.2%) patients had their operation within 24 h of presenting to hospital. The remaining 91 patients had a total of 239 days delay (in excess of the initial 24 h) for surgical treatment. Active medical problems (56.5%) and a wait for medical investigations (19.7%) caused most delays. Lack of operating theatre time and Sunday trauma lists caused 23.8% of delays. CONCLUSIONS: Medical problems account for most delays of surgical treatment of hip fractures. A multidisciplinary approach, with early input by medical and anaesthetic teams, is essential in managing such patients. Established protocols may reduce waiting times for essential investigations. PMID- 12648345 TI - The use of skull X-rays in the accident and emergency department. AB - The notes of all patients attending the accident and emergency department at the Royal Berkshire Hospital with a head injury from 1-30 September 1999 were analysed for the indications for skull X-ray, the report on film, and the outcome of the consultation. Using the existing Royal Berkshire Hospital guidelines, 50% (193/385) of all patients had skull X-rays performed. One fracture was detected. If the recent guidelines from The Royal College of Surgeons of England Working Party for the use of skull X-rays in institutions which possess a CT scanner were applied, the number of skull X-rays performed would reduce from 193 to 14 without detriment to any patient. PMID- 12648346 TI - Patient perceptions of MRSA. AB - Drug-resistant nosocomial infections are an increasing problem. This issue has received considerable media coverage. To our knowledge there have been no studies investigating patient awareness and perceptions of nosocomial infections. A total of 113 surgical out-patients completed a questionnaire stating whether they had heard of either superbugs or MRSA. A series of questions were asked about the source of any information; methods of transmission and prevention; the consequences of infection and their emotional response if they were to acquire MRSA. Fifty patients (44%) had heard of superbugs or MRSA mainly via the media (58%) or from hospital staff (44%). The majority would feel either angry or afraid if they acquired MRSA in hospital, but there was good awareness of both methods of infection control and the consequences of infection. From our study, we conclude that the media is at least as important as health professionals in providing information. Concerns regarding nosocomial infection may need to be addressed prior to admission. PMID- 12648348 TI - Controlling cement chips in revision arthroplasty. PMID- 12648347 TI - What to do if it gets 'bigger'. AB - The problems associated with intra-operative erections are discussed. Present theory is reviewed to allow a better understanding of the available treatment options. PMID- 12648349 TI - A novel use of a 'DHS-type' plastic vertical isolation drape in spinal surgery. PMID- 12648350 TI - Cable ties for pelvic stabilisation. PMID- 12648351 TI - A new way of monitoring and nursing patients with melaena. PMID- 12648352 TI - A simple technique to avoid inadvertent deep anchorage of surgical drains. PMID- 12648354 TI - Improving the view at flexible cystoscopy. PMID- 12648353 TI - Tear-drop incision for mastectomy to avoid dog-ear deformity. PMID- 12648355 TI - A do-it-yourself vascular anastomosis simulator. PMID- 12648356 TI - Surgical treatment of para-oesophageal hiatal hernia. PMID- 12648357 TI - Surgical site identification--think before you print! PMID- 12648358 TI - The Internet--friend or foe? A questionnaire study of orthopaedic out-patients. PMID- 12648360 TI - Issues of consent in renal transplantation. PMID- 12648361 TI - Breast augmentation should be on the NHS: a discussion of the ethics of rationing. PMID- 12648362 TI - Acute cholecystitis--room for improvement? PMID- 12648363 TI - Should ICU be part of surgical training? PMID- 12648365 TI - Six-fold suture: wound length ratio for abdominal closure. PMID- 12648367 TI - Surgical skills training in middle-ear surgery. AB - Until recently the practice of otological procedures on cadaver temporal bones was a common occurrence in otolaryngology departments. The difficulty in acquiring specimens has led to alternative techniques which involve artificial and computer-aided models. This article looks at the present situation in these rapidly developing areas and describes an artificial model developed by the senior author for training in middle-ear procedures. PMID- 12648368 TI - Simple snoring: current practice. AB - Snoring is a significant problem with both social and medical manifestations. This article addresses the current management of the simple snorer from initial out-patient assessment to definitive treatment. PMID- 12648369 TI - Bacterial aetiology of non-resolving otitis media in South African children. AB - Little is known of the aetiology, serotypes or susceptibility of the pathogens causing non-resolving otitis media in children receiving care from specialists in private practice in developed or in developing countries. Increased access to antibiotics in the community amongst children receiving such private care in South Africa may be anticipated to lead to levels of resistance similar to those found in countries with similar models of private practice, such as the United States. This study was conducted to determine the aetiology of non-resolving otitis media in South African children receiving private care and to determine the antimicrobial resistance patterns and serotypes of the bacterial isolates. Middle-ear fluid was cultured from 173 children aged two months to seven years with non-resolving acute otitis media accompanied by persistent pain or fever who were referred to otorhinolaryngologists for drainage of middle-ear fluid within 14 days of the start of symptoms. While 92 per cent of the children had recently received antibiotics and 54 per cent were currently receiving them, bacteria were isolated from 47 children (27 per cent). Streptococcus pneumoniae was the most common pathogen (35), followed by Haemophilus influenzae (nine), Staphylococcus aureus (six), Moraxella catarrhalis (two), Streptococcus pyogenes (two) and Pseudomonas aeruginosa (one). Two isolates were identified in each of eight children. Antimicrobial resistance to one or more antibiotics was found in 33/35 (94 per cent) of the pneumococci isolated, with resistance to penicillin in 86 per cent, resistance to trimethoprim-sulfamethoxazole in 54 per cent and to erythromycin and clindamycin in 69 per cent and 57 per cent, respectively. The pneumococcal serotypes found were 19F (28 per cent), 14 (26 per cent), 23F (23 per cent), 6B (nine per cent), 19A (87 per cent), and four (three per cent). Children with a bacterial pathogen isolated were younger (mean age of 17 months) than children from whom no bacteria were isolated (mean age of 23 months; p = 0.03). Isolation of a pneumococcus was also significantly associated with younger age (mean = 16 months versus 22 months, p = 0.03), the presence of fever (OR = 2.15, p = 0.049), and having one or more prior episodes of otitis media within the six months before tympanocentesis (OR = 7.72, p = 0.03). Almost all pneumococci isolated from non-resolving acute otitis media in this community are antibiotic-resistant and should be considered especially in young children who have failed previous therapy and who have non-resolving pain or fever. PMID- 12648370 TI - Failure of antibiotic therapy in acute otitis media. AB - The aim of this retrospective study was to determine the possible causes of failure of antibiotic therapy in children with acute otitis media (AOM). Thirty nine samples of middle-ear fluid were obtained by myringotomy from 31 children suffering from AOM, unrelieved by antibiotic therapy administered for over 48 hours. The samples were analysed by the usual microbiological techniques, including cultures, tests for beta-lactamase producing strains and the determination of the minimal inhibitory concentration of penicillin for Streptococcus pneumoniae. In 14 samples, no bacterial strains were detected in the cultures of middle-ear fluid; and in two samples the cultures revealed two strains of bacteria. The bacteria most frequently identified were Haemophilus influenzae, found in 11 samples, and Streptococcus pneumoniae, found in seven samples, of which four produced strains with reduced susceptibility to penicillin. The failure of antibiotic therapy in AOM appears to be related to the increased resistance of Haemophilus influenzae and to the reduced susceptibility of Streptococcus pneumoniae to penicillin. Other factors contributing to the failure of antibiotic therapy in AOM may be the viruses or the bacteria that produce multiple pathogens in the middle ear. PMID- 12648371 TI - Mastoiditis in children. AB - Fifty-two children with mastoiditis were treated at the Federal Government Services Hospital over a period of 15 years (from 1986 to 2000). The salient feature in these cases was the presence of a post-auricular swelling and mastoid abscess. Extensive cholesteatoma was seen in 80.7 per cent of cases and 19.3 per cent had granulations in the mastoid cavity. Significantly, all patients had a history of chronic ear discharge prior to abscess formation. These children belonged to poor communities, were malnourished and attended hospital in a state of emergency. The presentation and course of the disease confirms the aggressive and potentially serious nature of the pathology requiring early medical attention in order to avert impending complications. It appears that in our environment acute mastoiditis with abscess formation develops from chronic otitis media and the cholesteatoma. The surgical procedures conducted in these cases were radical and modified radical mastoidectomy. PMID- 12648373 TI - Informed consent for septal surgery: the evidence-base. AB - Informed consent is becoming an increasingly important issue in patient care. Inherent in this is the ability to accurately inform patients of the risks and complications involved in surgery. Nasal septal surgery is commonly performed by otolaryngologists and although often considered routine has a number of recognized complications that can have a significant effect on life quality. We carried out a review of the literature over the last 25 years and reviewed studies quoting rates of complications after septoplasty and submucous resection (SMR) using strictly applied criteria for study inclusion. There was a paucity of prospective data, especially for the cosmetic effects of septal surgery. Rates of septal perforation were two to eight per cent for SMR and 1.6-5.4 per cent for septoplasty. The incidence of cosmetic deformity was low for both procedures zero to one per cent. More well-designed studies are required to augment the current evidence base. While this does not replace the need for surgeons to audit their own results it provides evidence-based information to assist in the process of informed consent. PMID- 12648372 TI - Myringostapediopexy: surgical expectation. AB - Myringostapediopexy may occur as a result of incus erosion with medialization of the tympanic membrane and is recognized as often producing serviceable hearing. The technique may be used as part of tympanoplasty following either canal wall up or canal wall down surgery for chronic otitis media. The use of this type of reconstruction is influenced by the anatomy of the ear after disease excision. This review of the hearing levels associated with myringostapediopexy shows that there is a similar range of hearing level both for naturally formed as well as surgically fashioned myringostapediopexy. For both 'naturally formed' and following canal wall up surgery about 80 per cent of patients will have an air bone gap of 20 dB or less compared to 60 per cent of those who undergo canal wall down surgery. PMID- 12648374 TI - Analytical and clinical evaluation of CYFRA 21-1 by electrochemiluminescent immunoassay in head and neck squamous cell carcinoma. AB - This paper attempts to evaluate the clinical usefulness of CYFRA 21-1 as a serum tumour marker in patients with head and neck squamous cell carcinoma (HNSCC). The serum concentration of CYFRA 21-1 was measured utilizing a new electrochemiluminescent immunoassay (ECLIA) in 142 patients with HNSCC before and after treatment, 68 patients with benign tumours of the head and neck, and 50 healthy controls. Serum levels of CYFRA 21-1 in patients with HNSCC were significantly higher than those of benign tumours and healthy controls (p < 0.001). The diagnostic sensitivity and specificity of CYFRA 21-1 for HNSCC were 62 per cent and 100 per cent, respectively. The positive rates of CYFRA 21-1 increased with progression of HNSCC, serum CYFRA 21-1 levels were related to the tumour stage expressed by primary tumour (T) and nodal status (N) (p < 0.001), but not related to patient age, gender, smoking and drinking habit, or histopathological grade (p > 0.05). Post-treatment levels of CYFRA 21-1 in HNSCC decreased significantly (p < 0.001). Among 38 patients with clinical or radiological evidence of a recurrence during follow-up, 78.9 per cent (30 of 38) showed an increase in CYFRA 21-1. The analytical ECLIA performance for serum CYFRA 21-1 provides a new means of clinical assessment for HNSCC. The results of ECLIA suggest that the serum marker CYFRA 21-1 is valuable not only for diagnosis but also for close monitoring of patients with HNSCC. PMID- 12648375 TI - The Canterbury technique for canalplasty via an endaural approach in the surgical management of chronic refractory otitis externa. AB - We describe an endaural technique for soft tissue and bony meatoplasty and canalplasty in the surgical management of chronic otitis externa unresponsive to medical treatment and out-patient microsuction. Following analysis of the surgical outcome via a retrospective review of patient records, we would recommend this surgical treatment in this patient group. PMID- 12648376 TI - Sutureless thyroidectomy using electrothermal system: a new technique. AB - Bleeding is a dangerous complication of thyroid surgery. We describe a new technique of vessel and tissue sealing and bleeding control using an electrothermal sealer. The technique has some potential advantages over the current methods used for this purpose. PMID- 12648377 TI - Mucoepidermoid carcinoma arising from the eustachian tube and middle ear. AB - We report a case of mucoepidermoid carcinoma (MEC) originating from the eustachian tube and middle ear. A 31-year-old male who presented with otorrhoea and methicillin-resistant Staphylococcus aureus (MRSA) in the right ear was admitted to hospital due to cerebral infarction and deep vein thrombosis. After recovery, biopsies from a granulomatous mass found in the middle ear during operation for chronic otitis media revealed intermediate-grade MEC and a nasopharyngeal mass identified after surgery also revealed the same result. He received combined radiation therapy and chemotherapy and no residual or recurrent tumour was detected after two years of follow-up. PMID- 12648378 TI - Primary middle-ear lymphoma in a child. AB - The case of a five year old boy who presented with a lower motor neurone facial nerve palsy secondary to primary non-Hodgkin's lymphoma (NHL) of the middle ear is discussed. Any child who presents with a facial nerve palsy and conductive hearing loss requires thorough evaluation to exclude the possibility of temporal bone malignancy. PMID- 12648379 TI - Early diagnosis and stage-adapted treatment of Wegener's granulomatosis. AB - A case of Wegener's granulomatosis (WG) presenting with hearing loss and right facial nerve palsy is reported. The definitive diagnosis was based upon clinical data and serum cANCA and AECA detection. Early assessment of WG prevented surgical facial nerve decompression to treat a chronic otitis media complication. Immunosuppressive therapy with steroids, cyclophosphamide and methotrexate was required for relief of clinical symptoms and cANCA negativity as an expression of disease remission. The effectiveness of co-trimoxazole for preventing relapses of WG is discussed. PMID- 12648380 TI - Autoimmune sensorineural hearing loss: is it still a clinical diagnosis? AB - Inner ear involvement with sensorineural hearing loss (SNHL) has been reported in many autoimmune disorders including ulcerative colitis. The pathogenetic mechanism of hearing loss in ulcerative colitis is thought to be immune mediated. Diagnostic tests are being developed to identify inner ear autoantibodies, that may be the cause of such hearing loss. The only test that is currently available for clinical use is the Otoblot test. This, however, tests only for antibodies against bovine heat shock protein 70 which is only one of the many cross-reacting proteins against the inner ear in suspected immune-mediated hearing loss. The clinical response to steroid therapy is thus the mainstay in the diagnosis of immune-mediated hearing loss. This paper presents a series of patients with clinically suspected autoimmune hearing loss. Diagnostic assays for this condition are discussed along with a review of the recent advances in the pathogenesis and laboratory diagnosis of immune-mediated sensorineural hearing loss. PMID- 12648382 TI - Resection of a large benign pharyngo-oesophageal submucous mesenchymoma via laterocervical approach. AB - Two rare cases of benign submucosal pharyngo-oesophageal mesenchymoma are presented in this paper. One patient was treated by tumour removal via a combined thoracic and laterocervical approach and the other by resection through a laterocervical approach. The paper discusses the pathology and diagnosis of benign mesenchymomas. The authors suggest that for large tumours located in the pharynx and extending down the oesophagus without adhesion to the oesophageal wall, the laterocervical approach can avoid complications associated with the thoracic approach. The new technique caused less tissue damage and provided a quicker recovery and shorter hospitalization. PMID- 12648381 TI - Incomplete tracheal duplication associated with severe unilateral lung hypoplasia. AB - A rare case of incomplete tracheal duplication with severe unilateral lung hypoplasia is presented. Photo-documentation of the gross post-mortem specimens is presented so that the anatomical aspects of this unusual anomaly can be recognized and appreciated. Clinical information is presented in the hope that successful premorbid identification of potential complications of this anomaly could be made by future physicians. To our knowledge, this is the first reported case of pathologically-confirmed duplication of the trachea. PMID- 12648383 TI - Internal jugular vein thrombosis following ovarian hyperstimulation. AB - Superovulation therapy with in vitro fertilization (IVF) treatment may result in ovarian hyperstimulation syndrome and a hypercoaguable state. The site of thrombus formation is commonly in the upper venous extremities. One case of bilateral internal jugular vein thrombosis following ovarian hyperstimulation is described. The clinical presentation, investigations and management are discussed. PMID- 12648384 TI - Early radiation-induced malignant fibrous histiocytoma of the oral cavity. AB - With an incidence of less than 0.3 per cent, post-radiation sarcomas are rare malignant neoplasms with a very poor prognosis. On average, they occur after a latency period of at least 15 years following radiation therapy with doses ranging from 24 to 80 Gy. We present the case of a post-irradiation malignant fibrous histiocytoma (MFH) on the floor of the mouth in a 79-year-old male patient arising only five and a half years after radiation therapy. The primary tumour was classified as a well differentiated squamous cell carcinoma of the right rim of the tongue. Primary therapy was surgical resection of the tumour and post-operative radiation with 50 Gy. Five and a half years later, the patient developed a rapidly progressing MFH within the field of radiation. PMID- 12648386 TI - Effects of orthographic transparency on reading and phoneme awareness in children learning to read in Wales. AB - The relationship between the development of reading skills and the consistency of the orthography (writing system) is investigated in a study that examines reading acquisition in children living in Wales. Performance of children learning to read Welsh (a transparent alphabetic orthography) on tests of reading and phoneme detection was compared with the performance of children learning to read English (an opaque alphabetic orthography). The children were tested during their second year of formal reading instruction at school when they were aged between 5 and 6 years, and again one year later. The children learning to read in Welsh performed significantly better at reading both real words and nonwords than children learning to read in English. The English readers made fewer phonologically based reading errors. The Welsh readers also performed better on a phoneme awareness task. These findings support the claim that children learn to read more quickly in a transparent orthography, and provide further evidence that the consistency of the orthography influences the initial adoption of different strategies for word recognition. PMID- 12648387 TI - Mental imagery and creativity: a meta-analytic review study. AB - The reports of many creative individuals suggest the use of mental imagery in scientific and artistic production. A variety of protocols have tested the association between mental imagery and creativity, but the individual differences approach has been most frequently employed. This approach is assessed here through a range of meta-analytic tests. Database searches revealed 18 papers employing the individual differences approach that were subjected to a conservative set of selection criteria. Nine studies (1494 participants) were included in the final analyses. A marginal, but statistically significant, Fisher's Z-transformed correlation coefficient was revealed. Further analyses showed little difference between form and type of self-reported imagery and divergent thinking. Explanations for the failure to account for more than 3% of the variance in the data sets are discussed in the context of anecdotal reports, task validity, and design problems. PMID- 12648388 TI - The effects of orientation on detection and identification of facial expressions of emotion. AB - Signal detection procedures were used to examine the ability of participants to detect and label facial expressions of emotion in an upright or inverted orientation when the faces were rapid videotaped presentations. The detection and identification of facial expressions were remarkably accurate. In the upright orientation, the A' measure of sensitivity was above.9 for detection and identification of all six facial expressions of emotion. Sensitivity to inverted expressions was diminished for all emotions; however, the extent of the decline in sensitivity depended upon the specific facial expression. If the expression was difficult to detect or label in the upright orientation, the sensitivity score was lower in the inverted orientation. An assessment of the errors made in the detection and labelling process allowed a demonstration of the specific facial expressions that were confused in either the upright or inverted orientation. The assessment of sensitivity and analysis of the errors suggests that the nature of perceptual processing of some, but not all, facial expressions is changed by inversion. PMID- 12648389 TI - Do clever brains age more slowly? Further exploration of a nun result. AB - Recent epidemiological evidence suggests that individuals who have higher levels of mental ability in youth experience a slower cognitive decline as they grow old. In a sample of 3,263 Newcastle residents, average scores on a vocabulary test (Raven's 1965 'Mill Hill A') did not vary, while average scores on a test of fluid mental ability (the Heim, 1970, AH 4 (1) group intelligence test) sharply declined with age from 49 to 92 years. In young adults, Mill Hill A scores are good proxies for AH 4 (1) scores. This relationship allowed individuals' youthful AH 4 (1) test scores to be estimated from their current, unchanged, Mill Hill A scores so that age-related changes in AH 4 test scores over the adult life-span could be estimated and compared between high and low ability groups, men and women, and individuals of different levels of socio-economic advantage. The cross sectional estimated rate of age-related decline in general mental ability was found to be the same for people of all levels of ability and socio-economic advantage, and not to differ between men and women. PMID- 12648390 TI - The effect of item feedback on multiple-choice test responses. AB - This study examines the effects of item feedback on multiple-choice test responses. In the first experiment, a positive effect was predicted under instructions advising a penalty for errors; a larger beneficial effect on female scores was expected, given the (presumed) tendency of female participants toward omission when uncertain. Feedback effects were either negligible or negative, and the expected interaction effect with gender was not found. The second experiment was a high-powered replication of the feedback effect on errors, controlling for participants' ability. The discussion takes into account other evidence to state that recommendations of providing item feedback in the context of testing are neither theoretically nor empirically founded. PMID- 12648391 TI - 'There's no smoke without fire': are male ex-partners perceived as more 'entitled' to stalk than acquaintance or stranger stalkers? AB - In a stalking scenario, the prior relationship between and the gender of stalker and victim were systematically manipulated in order to judge culpability and consequences for the persons involved. Written vignettes were presented to 168 participants who responded via seven Likert scales. Stalker-victim relationship had three levels: ex-partner, acquaintance and stranger. In accordance with the 'Just World' hypothesis (Lerner, 1980), the victim was judged as having greater responsibility for the stalking when their harasser was an ex-partner or a prior acquaintance rather than a stranger, and police intervention was felt to be most necessary when the stalker was a stranger. Sex of stalker and victim was manipulated, and the following comparisons proved significant: when the perpetrator was male, bodily injury to the victim was seen as more likely and police intervention as more necessary than when the perpetrator was female; and male victims were viewed as more responsible for the scenario and as possessing greater powers to alleviate it. The Just World hypothesis and gender stereotypes provide a plausible account for these findings. Future research should determine whether criminal convictions show similar biases towards convicting male and stranger stalkers more often than female and ex-partner stalkers. PMID- 12648392 TI - Effort after meaning and the hedonic value of paintings. AB - Two experiments tested the prediction that providing the viewer with information about abstract and semi-abstract paintings, in the form of titles and descriptions, would increase the paintings' perceived meaningfulness and hedonic value. The meaningfulness prediction was supported in both experiments. The results of one experiment, using a between-participants design, failed to support the hedonic value prediction, but the results of the other, using a within participants design, supported the prediction. Failure to find an effect of information on hedonic value in other experiments may result from the use of between-participants designs that are relatively insensitive to the effects of different evaluative conditions. The results are discussed in the light of the 'effort after meaning' theory that part of the pleasure derived from looking at a painting stems from making a successful interpretation of it and picking up the artist's message. PMID- 12648394 TI - [Diagnostic criteria of nonalcoholic fatty liver disease]. PMID- 12648393 TI - 'Most advanced, yet acceptable': typicality and novelty as joint predictors of aesthetic preference in industrial design. AB - Typicality and novelty have often been shown to be related to aesthetic preference of human artefacts. Since a typical product is rarely new and, conversely, a novel product will not often be designated as typical, the positive effects of both features seem incompatible. In three studies it was shown that typicality (operationalized as 'goodness of example') and novelty are jointly and equally effective in explaining the aesthetic preference of consumer products, but that they suppress each other's effect. Direct correlations between both variables and aesthetic preference were not significant, but each relationship became highly significant when the influence of the other variable was partialed out. In Study 2, it was furthermore demonstrated that the expertise level of observers did not affect the relative contribution of novelty and typicality. It was finally shown (Study 3) that a more 'objective' measure of typicality, central tendency - operationalized as an exemplar's average similarity to all other members of the category - yielded the same effect of typicality on aesthetic preference. In sum, all three studies showed that people prefer novel designs as long as the novelty does not affect typicality, or, phrased differently, they prefer typicality given that this is not to the detriment of novelty. Preferred are products with an optimal combination of both aspects. PMID- 12648395 TI - [Diagnostic criteria of alcoholic liver disease]. PMID- 12648397 TI - [Effects of tea polyphennols on hepatic lipase activity in rabbits with fatty liver]. AB - OBJECTIVE: To investigate the relationship between hepatic lipase activity and fatty degeneration of hepatocytes and explore the effects of tea polyphennols (TP) on the changes of hepatic lipase (HL) activity in rabbits with fatty liver. METHODS: According to serum cholesterol and triglyceride (TC) levels, 19 rabbits were divided into fatty liver group (FL, n=6) fed with high cholesterol diet, TP group (n=7) fed with high cholesterol diet and 20mug/g/d tea polyphennols everyday orally, control group (n=6) fed with normal diet. After 8 weeks, the levels of serum TC, HL activity, HL activity and malondildehyde (MDA) in hepatic tissue were detected, and the pathomorphology of hepatic tissue were determined in all rabbits. RESULTS: The fatty degeneration of hepatocyts in FL group was more severe than that in TP and control group. The serum TC level in TP group (16.87 6.58) mmol/L was higher than that (1.11 0.82) mmol/L in control group (t=5.786, p<0.05), but lower than that (28.49 5.99) mmol/L in FL group (t=3.968, p<0.05). The serum low density lipoprotein-cholesterol level in Tp group (5.10 4.19) mmol/L also higher than that (0.71 1.14) mmol/L in control group (t=3.763, p<0.05), but lower than that (12.15 1.95) mmol/L in FL group (t=2.478, p<0.05). The number of positive dots presenting HL activity level in 100 square micron, hepatic tissue in TP group (3.24 0.17) was higher than that (1.76 0.10) in FL group (t=-3.153, p<0.05), but lower than that (4.14 0.05) in control group (t= 2.902, p<0.05). The levels of MDA in hepatic tissue in TP group (44.66 26.18) nmol/mg was significantly lower than that (75.58 29.88) nmol/mg in FL group (t=2.261, p<0.05), but no evidently different from that (43.64 16.95) nmol/mg in control group. The plasma HL activity was no difference among the three groups. CONCLUSION: The HL activity in hepatic tissue with fatty degeneration of hepatocytes was lower than that in normal liver. Tea polyphennols can increase HL activity in hepatic tissue and protect hepatocytes from fatty degeneration. PMID- 12648396 TI - [Change of serum endotoxin level in the progress of nonalcoholic steatohepatitis in rats]. AB - OBJECTIVE: To explore the role of endotoxin in the pathogenesis of nonalcoholic steatohepatitis (NASH). METHODS: Rat models of NASH were established by giving a fat-riched diet. These rats were sacrificed at the 4th, 8th, 12th, 16th and 24th weeks during the study. The other rats fed with normal diet were taken as normal controls at the same stage during the study. The blood of abdominal aorta was obtained and the levels of serum endotoxin, tumor necrosis factor-alpha (TNF-a), and interleukin-1 beta (IL-1 b) were measured. The expression of CD(14) and lysozyme in rats' livers were detected by immunohistochemistry. RESULTS: Rat models of NASH with liver fibrosis were established successfully. The levels of endotoxin in aorta blood of NASH rats increased significantly at the 24th week (0.23 EU/L 0.06 EU/L vs 0.15 EU/L 0.03 EU/L, t>2.179, p <0.05) while the expression of CD(14) increased from the 4th week, and the Kupffer cells expressing lysozyme were activated, then kept increasing activation through the study. In NASH rats, the levels of serum TNF-a increased from the 8th week (26.39 pg/ml 24.21 pg/ml vs 9.82 pg/ml 9.29 pg/ml, t>2.145, p < 0.05) and serum IL-1beta increased from the 16th week (23.76 pg/ml 21.81 pg/ml vs 6.25 pg/ml 2.98 pg/ml, t>2.145, p<0.05). CONCLUSION: Liver injury results from endotoxin existing in NASH rats which may play an important role in the pathogenesis of NASH by activating Kupffer cells and inducing the production of cytokines, such as TNF-a. PMID- 12648398 TI - [Clinicopathologic analysis of shock liver]. PMID- 12648399 TI - [A clinicopathological study on nonalcoholic steatohepatitis]. AB - OBJECTIVE: To observe the pathological and clinical characters of nonalcoholic steatohepatitis (NASH). METHODS: Liver biopsy tissues taken from 97 patients negative for common viral detection with serological test and immunohistochemistry as well as in situ hybridization, were observed by routine light microscopic examination. And the clinical data of patients with NASH was analyzed. RESULTS: Among the chronic hepatitis patients with unknown etiology, the detection rate of NASH was 15.5% (15/97). The characteristic lesions in NASH patients included macrovesicular steatosis in zone 3 of lobules, hepatocytes ballooning degeneration, lobules diffused with acute and chronic inflammation, and perisinusoidal fibrosis. Grading and staging according to Brunt's method, histological lesions could be accounted for G1S1 in 7 patients, G2S2 in 3 patients, G3S1 in 4 patients and G3S2 in 1 patient. There were 14 patients with mild to moderate elevation of transaminase, 10 with hyperlipemia, 8 with diabetes and 9 with fatty liver by ultrasonography. CONCLUSION: Nonalcoholic steatohepatitis is a common form of unknown etiology chronic liver disease with certain clinic-pathological features. PMID- 12648400 TI - [Clinical characters and CT findings of steatohepatitis in highland area]. AB - OBJECTIVE: To explore the relationship between the CT findings of steatohepatitis and the special environmental factors in highland area, such as anoxia and life styles. METHODS: CT scan was performed for 64 patients, who were 44 males and 20 females in Tibet with a 3500-meter elevation. RESULTS: Hypodensity in liver parenchyma and the invertion of liver density and spleen density were found in all the 64 patients. According to the degree and scope in their pathological changes, these patients were divided into three groups: severe 14, moderate 26, and mild 24.And diffused 43, sectional 14, regional 7 by CT findings. Among the 44 drinkers, 34 with more than 10 years were diffused by CT scan, the other 10 drinkers under 10 years, diffused 8, and sectional 2. In addition, the 27 steatohepatitis patients were all diffused by CT who were obese and had drinked more than 10 years. CONCLUSION: anoxia and drinking excessively in highland area are relevant to fatty degeneration in liver. PMID- 12648401 TI - [Therapy effects of fenofibrate on alcoholic fatty liver and drug-induced fatty liver in rats]. AB - OBJECTIVE: To investigate the fat decreasing effects of fenofibrate on alcoholic fatty liver and drug-induced fatty liver in rats. METHODS: Alcoholic fatty liver and drug-induced fatty liver rats models were established. The two kinds of rats with fatty liver were seperatedly divided into fenofibrate treatment group (80 mg/kg daily) and control group without treatment. Rats were killed after four weeks, then the levels of serum triglycerides (TG), total cholesterol (TC), high density lipoprotein (HDL) and malondialdehyde (MDA), hepatic lipase (HL), lipoprotein lipase (LPL) both in serum and liver tissue were measured according to the Test Kits. Histopathological changes in liver was dyed with HE and observed under light microscope. RESULTS: After treatment by fenofibrate, in the serum of rats with alcoholic fatty liver, the level of TG decreased significantly (1.07 mmol/L 0.06 mmol/L vs 1.56 mmol/L 0.29 mmol/L, t=5.115, p<0.001), while the level of TC had no alteration. The levels of MDA both in serum and liver tissue decreased (1.10 nmol/L 0.22 nmol/L vs 1.26 nmol/L 0.21 nmol/L, t=0.592, p<0.05; 5.92 nmol/g 1.24 nmol/g vs 7.42 nmol/g 1.22 nmol/g, t=3.477, p<0.05, respectively), while the levels of HL, LPL in serum and liver tissue increased significantly (Serum: 0.053muEq/ml/h 0.006muEq/ml/h vs 0.037 muEq/ml/h 0.006muEq/ml/h, t=-5.086, p<0.001; 0.018 muEq/ml/h 0.004 muEq/ml/h vs 0.014muEq/ml/h 0.004muEq/ml/h, t=-2.485, p<0.05. Liver tissue: 0.075muEq/ml/h 0.010muEq/ml/h vs 0.065muEq/ml/h 0.007muEq/ml/h, t=-2.437, p<0.05; 0.022 muEq/ml/h 0.014 muEq/ml/h vs 0.008 muEq/ml/h 0.002 muEq/ml/h, t=-2.876, p<0.05). Fat content in liver decreased (26.01 mg/g 1.69 mg/g vs 71.45 mg/g 2.66 mg/g, t= 43.224, p<0.001). The pathological changes of liver in fenofibrate-treated rats with alcoholic fatty liver were improved. For the drug-induced fatty liver rats, fenofibrate treatment group had no difference from the untreated control group. CONCLUSION: Fenofibrate can significantly decrease the fat content in liver tissue of rats with alcoholic fatty liver, as well as ameliorating liver pathological changes. But fenofibrate has no effect on drug-induced fatty liver. PMID- 12648402 TI - [Generation of high affinity humanized single-chain antibody against PreS1 of hepatitis B virus from immune phage displayed antibody library]. AB - OBJECTIVE: Screening and characterizing high affinity completely humanized single chain antibody (ScFv) against PreS1 of hepatitis B virus. METHODS: A combinatorial library of phage-displayed human ScFv, genes of which were derived from peripheral blood lymphocytes immunized by PreS1 of Hepatitis B Virus in vitro, was constructed. The library contained 7 10(8) clones. RESULTS: After 3 rounds panning, a high affinity (K=10(7) to 10(8) mol/L) ScFv specific to PreS1 was obtained. The V(H) belonged to human V(H4) family, and V(1) to V(4) by sequence analysis. CONCLUSION: This study suggests that the method of antigen stimulation in vitro is an expeditious way for the source of human immune antibody. And the ScFv may provide a more satisfactory therapy. PMID- 12648404 TI - [Mitochondrial microsatellite instability in hepatocellular carcinoma]. PMID- 12648403 TI - [Influence of Cordyceps sinensis on pancreatic islet beta cells in rats with experimental liver fibrogenesis]. AB - OBJECTIVE: To investigate the effect of Cordyceps sinensis (CS) on pancreatic islet B cells of experimental hepatic fibrogenesis rats. METHODS: Rats were randomly allocated into three groups: normal group, model group and CS group. The rats in the latter two groups were administered with CCl(4) solution to induce liver fibrosis, the CS group was also treated with CS 10 days after the beginning of CCl(4) administration. Rats in normal group were sacrificed at the beginning of the experiment, while the rats in the other two groups were sacrificed randomly at the end of the third and sixth weeks. The rats' islets were isolated and cultured in vitro, then the basal insulin level of the islets and the serum level of insulin were determined by radioimmunological assay. RESULTS: It seemed no change that the levels of serum insulin and basal insulin between the model group and the normal group at the third week. But at the sixth week, both insulin levels in the model group were higher than those in the normal group (52.6 mU/L2.5 mU/L vs 23.7 mU/L 2.3 mU/L, q=13.01, p<0.05; 52.94muU/ml 13.12muU/ml vs 35.16muU/ml 5.64muU/ml, q=10.06, p<0.01). No significant change could be seen in the serum levels of insulin between the CS group and the model group at the third and sixth weeks. But the basal insulin levels in the CS group were apparently higher than those in the model group at the third and sixth weeks (156.63muU/ml 6.57muU/ml vs 39.64muU/ml 3.95muU/ml, q=66.94, p<0.001; 140.44muU/ml 38.53muU/ml vs 52.94muU/ml 13.12muU/ml, q=12.98, p<0.01). CONCLUSION: Cordyceps sinensis can increase the basal insulin level of the islets in CCl(4)-induced liver fibrosis rats. PMID- 12648405 TI - [Different gene expression during hepatocarcinogenesis in tree shrew induced by aflatoxin B1]. AB - OBJECTIVE: To understand the molecular mechanism and find out the responsible genes for liver cancer by exploring the regulation of gene expression during hepatocarcinogenesis in tree shrew induced by aflatoxin B1 (AFB1). METHODS: The tissues from tree shrew of different stages during the pathogenesis and development of hepatocellular carcinoma (HCC), liver cancer tissue, para cancerous tissues, pre-cancerous liver tissues, liver tissues of the same stage from normal controls and the liver tissues taken before AFB1-treatment were analyzed for gene expression by cDNA array. RESULTS: Four patterns of gene expression were observed during AFB1-induced hepatocarcinogenesis. They were: genes up-regulated in HCC tissue and para-cancerous tissue, especially in HCC tissues; genes with similar expressing level in both HCC tissue and para cancerous tissue, but higher than that in pre-cancerous tissue; genes down regulated in HCC tissue; genes up-regulated before HCC appeared but down regulated after HCC appeared. CONCLUSION: Dynamic observation of gene expression will be beneficial to elucidate the mechanisms of AFB1- induced hepatocarcinogenesis and locate the responsible genes. PMID- 12648406 TI - [Expression characters of ATP7B mRNA in liver tissue of patients with Wilson's disease]. PMID- 12648407 TI - [Preparation and application of the monoclonal antibody against hepatoma-specific gamma-glutamyltransferase]. AB - OBJECTIVE: To prepare a monoclonal antibody against hepatoma-specific gamma glutamyltransferase (GGT-II) and study it's application. METHODS: Two Bal B/C mice were immunized with pure GGT-II, then their spleen cells were separated and fused to SP 2/0 myeloma cells so as to make hybridoma cell strain which could yield monoclonal antibody against GGT-II. And it's effect of binding GGT-II was detected by competitive inhibitory enzyme linked-immunosorbance assay (ELISA). RESULTS: A mouse hybridoma cell strain which could steadily secrete the monoclonal antibody against GGT-II was obtained and named 2G4F6B2. This monoclonal antibody belonged to IgG1 subclass and was specific to GGT-II, without cross-reaction to GGT-II. The result of detecting human serum GGT-II by ELISA with the monoclonal antibody accorded with that by polyacrylamide gradient electrophoresis. CONCLUSION: The monoclonal antibody against GGT-II prepared in this study has high specificity and can be applied in clinic to detect human serum GGT-II. PMID- 12648408 TI - [Effects of interferon on interferon sensitivity determining region 2209 aa~2248 aa in HCV NS5a]. PMID- 12648410 TI - [Effect of single-using or combined-using lamivudine/interferon-alpha 1b on serum markers in patients with hepatitis B]. PMID- 12648409 TI - [Chronic hepatitis B treated with domestic manufactured lamivudine in 2200 patients: a phase IV study]. AB - OBJECTIVE: To further verify the efficacy and safety of locally manufactured lamivudine on patients with chronic hepatitis B (CHB). METHODS: 2200 patients with CHB were recruited and received lamivudine orally 100 mg once daily for 12 months. The efficacy assessments included virologic response rate (defined by the absence of serum HBV DNA, HBeAg loss and HBeAg/HBeAb seroconversion), percentage of patients with normalization of alanine aminotransferase (ALT). Meanwhile improvement of quality of life (QOL) measured by mos SF-36 QOL questionnaire and liver histology evaluation were conducted in some patients. The safety assessments included adverse events, serious adverse events and laboratory abnormalities. All 2200 patients received at least one dose of medication and were all included in the safety population. RESULTS: Ninety seven percent of patients (2137/2200) recruited were HBV DNA positive by dot blot (sensitivity GRT or equal to 1.0 pg/ml) at baseline. At the end of 12 months treatment, HBV DNA was undetectable in 80% patients (1538/1920) with HBV DNA positive before treatment. Among the 79%(1744/2200) of the patients recruited had positive HBV DNA accompanied abnormal ALT levels at baseline, 72% patients became ALT normal. And among the 84% (1843/2200) of the patients recruited were HBV DNA and HBeAg positive, anti-HBe negative, 16% (269/1650) patients achieved HBeAg/HBeAb seroconversion after 12 months of lamivudine treatment. The HBeAg/HBeAb seroconversion rate was positive correlation to the ALT level before treatment. A total of 304 patients completed the health-related QOL questionnaire. After 12 months treatment, lamivudine improved both their physical and mental health, especially for their mental health. 133 evaluable, paired liver biopsies were obtained for histological assessment, among whom 115 patients had abnormal ALT levels at baseline. Compared with pre-treatment most of their liver injury got alleviated (51.9%) or no further deterioration (36%), only 12% worsening. During the 12 months treatment, 9% patients withdrew from the study and 17% patients showed at least one adverse event, mild or moderate. There were no obvious difference between this study and the previously reported lamivudine Phase II or III study with regard to the kinds, incidence and severity of adverse events. CONCLUSION: The efficacy and safety profile of the locally manufactured lamivudine 100 mg tablets are similar with those of the previously reported available lamivudine tablets imported in treating Chinese chronic hepatitis B patients. PMID- 12648411 TI - [Therapeutic strategies for nonalcoholic fatty liver disease]. PMID- 12648412 TI - [Treatment and prevention effects of weight loss on obese patients with nonalcoholic fatty liver disease]. PMID- 12648413 TI - [The role of improving insulin-resistant in the treatment and prevention of nonalcoholic fatty liver disease]. PMID- 12648414 TI - [Drug therapy for nonalcoholic fatty liver disease]. PMID- 12648415 TI - [Activation of nuclear factor-kappa in concanavalin A-induced mice liver injury]. PMID- 12648416 TI - [Genetic polymorphism of cytochrome P450II E1 related to the development of nonalcoholic fatty liver]. PMID- 12648417 TI - [Therapeutic effect of alpha 1b interferon on patients with chronic hepatitis B: changes in serological fibrosis markers and histology]. PMID- 12648418 TI - [Clinical analysis of hepatolenticular degeneration in children]. PMID- 12648419 TI - [Characteristics of free calcium distribution and Ca(2+) oscillations in the subregeon of hepatocytes]. PMID- 12648420 TI - [Current therapy strategies for nonalcoholic fatty liver disease]. PMID- 12648421 TI - [Mitochondrial reactive oxygen species and fatty liver]. PMID- 12648422 TI - [The role of cyclooxygenase-2 in the pathogenesis of alcoholic liver diseases]. PMID- 12648423 TI - [Classification and diagnostic strategies of nonalcoholic fatty liver diseases]. PMID- 12648424 TI - Are urologists in danger of becoming "technicians"? PMID- 12648425 TI - Preparedness in robotically assisted interventions. AB - For many years, robots have been used in manufacturing to perform a variety of delicate tasks. Their use is now being generalized to other fields, such as biology, domestic applications, and especially medicine, in which they are poised to make a significant contribution. This evolution comes from the progress made in the field of robotics and from recent changes in medical and surgical techniques, namely, developments in medical imaging and a new desire for minimally invasive interventions. This emerging combination of high-precision robotic manipulators, new medical diagnostic techniques, and efficient minimally invasive surgery has not yet been perfected. After a brief discussion of state-of the-art robotic systems used in urology, this article discusses new challenges presented by robotic minimally invasive surgery. A computer-integrated approach aimed at increasing the efficiency of such interventions through better preparedness is presented. This approach is illustrated by a case study in human nephrectomy and a cardiac animal experiment. PMID- 12648426 TI - Telesurgery, telementoring, virtual surgery, and telerobotics. AB - With the advent of laparoscopic surgery, a method characterized by a surgeon's lack of direct contact with the patient's organs and tissue and the availability of magnified video images, it has become possible to incorporate computer and robotic technologies into surgical procedures. Computer technology has the ability to enhance, compress, and transmit video signals and other information over long distances. These technical advances have had a profound effect on surgical procedures and on the surgeons themselves because they are changing the way surgery is taught and learned. This article provides an overview of the most important advances and issues developing from the use of computer and robotic technologies in surgery. PMID- 12648428 TI - Vattikuti Institute prostatectomy: surgical technique and current results. AB - The Vattikuti Urology Institute has been developing a robotic prostatectomy system for the management of prostate cancer. This technique is based on the scientific foundations of Walsh's anatomic prostatectomy. Two hundred fifty patients with clinically localized prostate cancer have undergone this technique. Preoperative, operative, and postoperative parameters were collected, and functional outcomes using previously validated quality-of-life instruments were evaluated. The mean operating time for these patients was 2.5 hours (165 and 135 minutes with and without lymphadenectomy, respectively), and the average blood loss was 150 mL. The median specimen Gleason score was 7, and the mean tumor volume was 7 mL. Four patients had a positive surgical margin (three unifocal, one multifocal). Ninety-five percent of the patients were discharged within 23 hours, and the mean catheterization time was 4.2 days. The complication rate was 4%. Approximately 78% of the patients had intact erectile response and 96% achieved continence by the sixth month after surgery. The robotic prostatectomy system is a safe and effective operation for the management of prostate cancer. PMID- 12648427 TI - Principles and advantages of robotics in urologic surgery. AB - Although the available minimally invasive surgical techniques (ie, laparoscopy) have clear advantages, these procedures continue to cause problems for patients. Surgical tools are limited by set axes of movement, restricting the degree of freedom available to the surgeon. In addition, depth perception is lost with the use of two-dimensional viewing systems. As surgeons view a "virtual" target on a television screen, they are hampered by decreased sensory input and a concurrent loss of dexterity. The development of robotic assistance systems in recent years could be the key to overcoming these difficulties. Using robotic systems, surgeons can experience a more natural and ergonomic surgical "feel." Surgical assistance, dexterity and precision enhancement, systems networking, and image guided therapy are among the benefits offered by surgical robots. In return, the surgeon gains a shorter learning curve, reduced fatigue, and the opportunity to perform complex procedures that would be difficult using conventional laparoscopy. With the development of image-guided technology, robotic systems will become useful tools for surgical training and simulation. Remote surgery is not a routine procedure, but several teams are working on this and experiencing good results. However, economic concerns are the major drawbacks of these systems; before remote surgery becomes routinely feasible, the clinical benefits must be balanced with high investment and running costs. PMID- 12648431 TI - Improved survival in relapsed Wilms' tumor in the modern era. PMID- 12648429 TI - Robotic pyeloplasty. AB - In addition to the classic open surgery, a variety of minimally invasive therapeutic options have been developed for the treatment of ureteropelvic junction obstruction, including an endoscopic antegrade or retrograde ureteropelvic junction obstruction visually controlled incision or radioscopically controlled Acucise (Applied Medical, Laguna Hills, CA), which does not share the high success rate that results from open-surgical dismembered pyeloplasty. Laparoscopic pyeloplasty, which duplicates the open technique and differs only by the mode of access, has proven to have positive results when performed by experts, but remains a demanding technique that requires a long learning curve. Providing a three-dimensional vision, an unprecedented control of the endocorporeal instruments, and an ergonomic surgeon's position, robots may allow urologists with limited laparoscopic experience to rapidly master the endocorporeal management of ureteropelvic junction obstruction. They likely will propel minimally invasive urology forward in the next several years. PMID- 12648432 TI - Biofeedback therapy and dysfunctional voiding in children. AB - Dysfunctional voiding, a condition in which a neurologically intact child fails to relax the urinary sphincter during micturition, was first described in the 1970s. Clinically, these children have urinary incontinence and recurrent urinary tract infections. Biofeedback, through which the child is taught to relax the pelvic floor during voiding, has become an increasingly popular method of treatment. Many series, most retrospective, have shown biofeedback to be very effective in the treatment of this disorder. PMID- 12648430 TI - Laparoscopic and robotic surgical training in urology. AB - The most important change in urology during the past decade was the development of minimally invasive surgery, particularly laparoscopy. However, the main drawback of laparoscopy is a steep learning curve, which results from the significant changes in the surgical environment. Although laparoscopy can provide important advantages for the patient, including decreased length of hospitalization, decreased analgesic requirement, and a shortened postoperative convalescence, one concern has been whether laparoscopic techniques should be learned solely in the operating room. For example, sports, music, and aviation are practiced before an actual performance is ever undertaken. In this review, the advantages and limitations of all available training modalities in minimally invasive surgery are described. Testing basic laparoscopic skills on inanimate models, becoming familiar with the principles of dissection and hemostasis on living animals, and studying surgical anatomy on cadavers should be considered as indispensable and complementary elements for laparoscopic training in the future. In addition, telementoring with the help of modern image processing and virtual reality eventually may become the basis of tomorrow's surgical instruction. PMID- 12648433 TI - Urologic anomalies and two-vessel umbilical cords: what are the implications? AB - A two-vessel umbilical cord (or single umbilical artery) is a perinatal finding that may be detected in infants with congenital or chromosomal abnormalities. Its presence has been associated with an increased risk of genitourinary defects. The urologic anomalies that typically are associated with a two-vessel cord often are minor and self-limiting. These defects usually can be detected perinatally in routine prenatal screening ultrasonographic examinations. The neonatal physical examination, an additional postnatal ultrasonography, and possibly a voiding cystourethrogram will be sufficient to complete the screening for genitourinary abnormalities in children with a single umbilical artery. Infants with two-vessel umbilical cords do not require additional screening for genitourinary defects, unless the imaging or amniocentesis detects a genitourinary abnormality. This review details the history of two-vessel cords and the associated genitourinary anomalies. PMID- 12648434 TI - Two-stage repair for proximal hypospadias: a reappraisal. AB - Great advances have been made in surgery for the correction of hypospadias. Proximal hypospadias remains the greatest challenge, but, despite many innovations and much progress, surgery can fail. Many authorities have introduced single-stage techniques, which have the purported advantage of correcting the defect with minimal hospitalization and family inconvenience. However, the wider published experience with these approaches would suggest that the ideal single stage procedure has yet to be devised. A substantial number of children undergoing a single-stage procedure will have to undergo further surgery. A small, but irreducible number of patients will be rendered hypospadias "cripples." A number of these children will be dissatisfied as adults because of cosmetic or functional deficiencies. A two-stage approach for correction of proximal hypospadias and severe chordee remains the most credible and reliable solution for many of these patients. Pediatric urologists should maintain familiarity with these techniques and continue to advocate them for a select group of patients. PMID- 12648435 TI - Tissue engineering in urology: where are we going? AB - Tissue engineering in urology is a broad term used to describe the development of alternative tissue sources for diseased or dysfunctional native urologic tissue. This article reviews the recently published techniques involving synthetic and natural biodegradable matrices alone, known as "unseeded" scaffolds, and the latest data on "seeded" scaffolds, which are impregnated with cultured cells from urologic organs. Recent discoveries in reporter gene labeling of urologic tissue are discussed as a new method to identify and track the fates of these transplanted cells in vivo. This article also investigates how these bioengineering techniques are applied to synthetic and natural scaffolds, such as polyglycolic acid and porcine small intestine submucosa, to increase bladder capacity, repair urethral strictures, and replace corporal plaques in Peyronie's disease. Furthermore, recently published reports that these materials have been seeded with chondrocytes to create corporal rods for penile prostheses and stents for ureteral and urethral stricture disease are discussed. With these latest developments as a foundation, the future directions of tissue engineering in urology are presented. PMID- 12648436 TI - Molecular mechanisms of renal development. AB - The biology of renal development has become increasingly complex because technical advances in genetics and cell biology have been used to study this aspect of embryogenesis. The molecular biology and genetics of renal development may seem inconsequential and frustrating to the practicing clinician, but insight into fundamental mechanisms of renal development are necessary to understand clinical breakthroughs that will occur in the future. As a basis for appreciating these concepts, specific paradigms of renal development are illustrated and the investigative strategies used to develop them are summarized in this article. PMID- 12648437 TI - Molecular genetics of renal development. AB - The understanding of the complex branching morphogenesis of the early kidney is at an early stage; however, a framework is emerging that suggests numerous active genes sequentially switching on and reciprocally influencing each other. Much of our understanding of this process comes from studies of rodents specifically engineered to lack a particular gene responsible for an inductive agent or receptor. This review attempts to place newly discovered genetic programs within an organized framework of sequential renal development. PMID- 12648438 TI - Preeclampsia is associated with a reduced interleukin-10 production from peripheral blood mononuclear cells. AB - PROBLEM: This study aims to investigate and compare in vitro, immune cell production of the immunosuppresor cytokine interleukin-10 (IL-10) and the proinflammatory cytokine tumor necrosis factor (TNF-alpha) between normal pregnancy (NP) and preeclampsia. METHODS: Peripheral blood mononuclear cells (PBMC) were isolated from age-matched patients with preeclampsia and women with a NP (n=3/group) and cultured for 48 hr in the absence and presence of the mitogen phytohaemagluttanin (PHA, 1/100). The concentration of IL-10 and TNF-alpha in the culture medium (CM) was measured by ELISA. RESULTS: Stimulated PBMCs associated with preeclampsia had significantly less IL-10 in the CM compared to NP (347+/ 39, preeclampsia vs. 689+/-128, NP; p<0.05) but no difference in TNF-alpha. There was no significant difference in IL-10 or TNF-alpha concentration in the CM of unstimulated PBMCs between preeclampsia and NP. CONCLUSIONS: These findings suggest that under in vitro stimulated conditions preeclampsia is associated with an abnormality characterized by a diminished ability of peripheral immune cells to produce the immunosupressor cytokine interleukin-10. PMID- 12648440 TI - Neurodevelopmental and cognitive assessment of children born growth restricted to mothers with and without preeclampsia. AB - We examined neurological and intellectual outcome of growth-restricted newborns of pregnancies complicated with preeclampsia and without preeclampsia. Seventy five consecutive growth restricted newborns (<5th percentile) were prospectively followed up at 6 months' intervals. Newborns with major congenital malformations and newborns with evident intrauterine viral infection were excluded. At 3 years of age all children had detailed neurological examination and intellectual examination using the Mean developmental index (Stanford Binnet-IQ). Eleven children were born to mother with preeclampsia (ACOG criteria), and 64 were born to mothers without a definite diagnosis of preeclampsia. Gestational age was 34.7 weeks in the preeclamptic group and 37 weeks in the non-preeclamptic group. After adjustment for gestational age, there was no significant difference in the neurological exam score between groups, but the IQ was 85.5 in the preeclamptic group and 96.9 in the non-preeclamptic group (p<0.03). We conclude that newborns born growth restricted after pregnancies complicated by preeclampsia have a lower IQ at the age of 3 years compared to growth-restricted babies without preeclampsia. PMID- 12648439 TI - Adrenomedullin requires an intact nitric oxide system to function as an endogenous vasodilator in rat gestation. AB - OBJECTIVE: We hypothesize that administration of adrenomedullin (AM), an endogenous vasodilator, will ameliorate the hypertension and growth restriction associated with chronic nitric oxide inhibition induced by L-omega nitro-L arginine methyl ester (L-NAME) infusion in pregnant rats. METHODS: Osmotic minipumps were inserted on day 14 of gestation to deliver continuously either AM, L-NAME, AM+L-NAME, or vehicle control. Systolic blood pressure was recorded daily in pregnant rats. Pregnant rats were either sacrificed on gestational days 15, 16, 17, 18, or 22, or they were allowed to deliver at term. The placentas from all of the treated groups were fixed for 24 hr in Bouin solution, sectioned, processed, embedded in paraffin, and stained with hematoxylin and eosin. The placentas were graded for the quality of fetal vessel development in the labyrinth. RESULTS: Systolic blood pressure was increased in AM+L-NAME-treated rats. The animals that delivered in the AM+L-NAME group exhibited decreased pup weight (L-NAME and AM+L-NAME, 5.2+/-0.1 compared with 6.4+/-0.1 g for both AM and controls, p<0.001) and increased pup mortality (AM+L-NAME, 44.4% compared with 16.7% in L-NAME, 0% in AM and 3.1% in controls, p<0.001 AM+L-NAME compared with controls). Increased decidual necrosis, necrosis in the labyrinth, and deficient fetal vessel development in the labyrinth was identified in the placentas treated with AM+L-NAME. CONCLUSIONS: Addition of the endogenous vasodilator AM to an L NAME-induced state of chronic NO inhibition did not ameliorate hypertension and growth restriction. PMID- 12648441 TI - Elevated levels of adhesion molecules derived from leukocytes and endothelial cells in patients with pregnancy-induced hypertension. AB - OBJECTIVE: Our purpose was to elucidate the role of adhesion molecules in the pathogenesis of pregnancy-induced hypertension (PIH). METHODS: Sera, peripheral lymphocytes, and polymorphonuclear cells (PMN) from PIH patients, normal pregnant women, and nonpregnant women were collected. Soluble intercellular adhesion molecule-1 (sICAM-1) in sera was measured by ELISA. ICAM-1 expression on endothelial cells (EC) incubated with sera was analyzed by flow cytometry and RT PCR. CD11a, CD11b, and CD18 expression on lymphocytes and PMN were also measured by flow cytometory. RESULTS: CD11a and CD18 expression levels on PMN and lymphocytes of PIH patients were significantly higher than those of normal pregnant women (p<0.05). The expression of CD11b was significantly increased in normal pregnancy compared with that in nonpregnant women (p<0.05). Serum sICAM-1 in PIH patients was higher than that in normal pregnant women (p<0.05). ICAM-1 expression level on EC incubated with PIH serum for 24 hr was significantly higher than that with normal pregnant serum (p<0.0005). ICAM-1 mRNA expression after 12-hr incubation with PIH serum was also significantly increased compared with serum from normal pregnant women (p<0.05). CONCLUSION: Adhesion molecules may play an important role in the pathogenesis of PIH. PMID- 12648442 TI - Women with preeclampsia have increased serum levels of pregnancy-associated plasma protein A (PAPP-A), inhibin A, activin A and soluble E-selectin. AB - OBJECTIVE: Poor placentation in early pregnancy is thought to lead to an excessive maternal systemic inflammatory response, which causes the maternal syndrome of preeclampsia. The aims of this retrospective study were to confirm old reports of increased blood levels of pregnancy-associated plasma protein A (PAPP-A) in preeclampsia and how its levels correlate with the levels of other placental and endothelial proteins that are reported to be elevated in preeclampsia. METHODS: Nineteen women with preeclampsia symptoms were matched with 19 normal pregnant controls for gestational age, maternal age, and parity. PAPP-A, placental pregnancy-specific beta1-glycoprotein (SP1), inhibin A, activin A, and sE-selectin were measured in serum using specific ELISAs. RESULTS: Maternal serum levels of PAPP-A, inhibin A, activin A and sE-selectin were increased in women with preeclampsia (mean 157.7 vs. 76.85 mIU/mL, p=0.005; 3.08 vs. 1.51 ng/mL, p=0.002, 32.36 vs. 3.77 ng/mL, p<0.001 and 62.15 vs. 46.37 ng/mL, p=0.02 respectively), compared to controls. Serum levels of SP1 were not altered in preeclampsia. PAPP-A (r=0.636, p<0.01) had a positive correlation with sE selectin in patients with preeclampsia. Serum inhibin A and activin A had a significant positive correlation with each other in preeclampsia. CONCLUSIONS: Raised levels of PAPP-A in preeclampsia confirm earlier reports. Activin A showed the highest increase over the controls and is thus likely to be a better serum marker for this pathology than the other markers that were tested. PMID- 12648443 TI - Correlation of platelet count changes with liver cell destruction in HELLP syndrome. AB - OBJECTIVE: HELLP syndrome is defined as a subset of preeclampsia involving a combination of hemolysis, low platelets affecting the circulatory system, and elevated liver enzymes, reflecting liver cell destruction. Our objective was to assess the correlation between the platelet count changes and liver cell destruction as indicated by aspartate aminotransferase (AST) levels. METHOD: We reviewed the records of 15 preeclamptic patients who developed HELLP syndrome as defined by 1) hemolysis, 2) elevated liver enzymes with an AST greater than 70 U/L (>2 SD above mean), and 3) low platelets (<100x10(9)/L). In these patients, both platelet counts and AST were assessed every 4-6 hr before and after delivery until recovery. At least eight laboratory assessments were done on each patient and were available for review. In each patient, the correlation between platelet count and AST levels was done using the Spearman rank order correlation coefficient. RESULTS: A negative correlation was found between platelet count and AST levels. A correlation coefficient of greater than 0.8 occurred in 53% of cases. The correlation coefficient ranged from 0.1 to 0.93. In all cases, improvement of liver cell dysfunction occurred within 4 hr of recovery of platelet count. At 72 hr postpartum, 73% of the platelet counts had not recovered to >150x10(9)/L, and 66% of patients with abnormally elevated AST levels had not returned to <35 U/L. CONCLUSIONS: Liver dysfunction and platelet count are poorly correlated in individual cases. This lack of correlation is evident both during decline and recovery of the platelet counts. Changes in platelet count cannot be used as a proxy for liver dysfunction in severe preeclampsia with HELLP syndrome. PMID- 12648444 TI - Nitric oxide production by decidual endothelial cells is not reduced in preeclampsia. AB - OBJECTIVE: To investigate whether decidual endothelial cells (DEC) contribute to the pathogenesis of preeclampsia through abnormal nitric oxide production. Decidual endothelial cells from normal (NDEC) and preeclamptic (PEDEC) pregnancies, and also human umbilical vein endothelial cells (HUVEC), were examined. METHODS: HUVEC, NDEC, and PEDEC were incubated for 45 min in serum-free media with the addition of potential stimulators [calcium ionophore (A23187), sepiapterin, and a combination of cytokines (TNF-alpha, gamma-IFN and LPS)], and the competitive inhibitor, NG-monomethyl-L-arginine (L-NMMA). These were added alone or in combination. Supernatants were measured for nitrate/nitrite (NOx) levels and the cells acid-extracted for measurement of cyclic guanosine monophosphate (cGMP). The effect of 30 min of shear stress (approximately 20 dynes/cm2) on NO and cGMP production by NDEC and PEDEC and on production of prostacyclin and thromboxane A2, was assessed. RESULTS: PEDEC and HUVEC both produced more NO than NDEC under all conditions examined. Cell-associated cGMP levels, however, were not different among the cell groups but were increased by A23187 and inhibited by L-NMMA. In control conditions, shear stress stimulated cGMP levels 5-fold (p<0.01) in both NDEC and PEDEC, and PGI2 production 2-fold (p<0.05). CONCLUSIONS: DEC from preeclamptic women do not have reduced NO production and respond normally to shear stress by increasing cGMP and PGI2 production. Our results are consistent with other reports of equal or higher NO levels in preeclampsia and indicate that reduced NO production by endothelial cells is not the explanation for the vasoconstriction of uterine vessels. PMID- 12648445 TI - Validation of the DCA 2000 microalbumin:creatinine ratio urinanalyzer for its use in pregnancy and preeclampsia. AB - OBJECTIVE: To determine the accuracy of the DCA 2000 albumin/creatinine ratio urinanalyzer (Bayer Corp., Elkhart, IN) in uncomplicated pregnancy and preeclampsia. METHODS: This was a prospective observational study in a large teaching maternity hospital. Ninety one uncomplicated pregnant women and 100 women referred for assessment of de novo hypertension in pregnancy had albumin concentrations, creatinine concentrations, and albumin/creatinine ratios (ACR) compared between the DCA 2000 and the laboratory gold standard assays (Dade Dimension clinical chemistry autoanalyzer), for both early morning urines (EMU) and 24-hr urine collections. RESULTS: The interassay and intra-assay variability for the DCA 2000 were less than 5.1%. In uncomplicated pregnancy the mean difference in ACR between the DCA 2000 and the laboratory assay was 0.08 mg/mmol (SD 0.28; 95% limits of agreement, -0.47, 0.63) for EMU and 0.06 (SD 0.23; 95% limits of agreement, -0.39, 0.51) for 24-hr samples. In the hypertensive cohort the ACR mean differences were -0.82 (SD 7.13; 95% limits of agreement, -14.79, 13.15) for EMU and -0.76 (SD 4.14; 95% limits of agreement, -8.87, 7.35) for 24 hr samples. The mean differences between assays in the hypertensive group had broader 95% limits of agreement due to greater variability in the samples with high albumin concentrations (>40 mg/L). CONCLUSIONS: The DCA 2000 is accurate for the measurement of albumin creatinine ratios in the uncomplicated pregnant population. In the hypertensive population the DCA 2000 remains accurate though when the albumin concentration is greater than 40 mg/L the 95% limits of agreement are broader. We would recommend that all other automated urinalysis devices be validated by similar protocols to allow meaningful comparisons of accuracy. PMID- 12648446 TI - Patient initiated home blood pressure recordings are accurate in hypertensive pregnant women. AB - OBJECTIVE: We undertook this study to determine the accuracy and reliability of patient initiated blood pressure measurement and recording. METHODS: We recruited 72 women from the antenatal hypertension clinic in a university teaching hospital. All were at high risk for preeclampsia and were asked to measure and record their blood pressure three times per day at home using a validated blood pressure device with an internal memory. RESULTS: From 979 measurements taken only 28 (2.9% were inaccurate). The inaccurate readings were restricted to three women. On further questioning two women admitted that the device had been used by other family members thus making comparison with the memory impossible. Thus the true nonconcordance rate amongst participants was 1/72 (1.4%). CONCLUSIONS: We conclude that blood pressure recordings taken and documented by high-risk women at home are accurate. This allows more frequent measurements to be taken without the inconvenience of additional visits to hospital and may therefore lead to the earlier detection of preeclampsia. PMID- 12648447 TI - Increased endothelial monolayer permeability is induced by serum from women with preeclampsia but not by serum from women with normal pregnancy or that are not pregnant. AB - OBJECTIVE: To determine if endothelial monolayer permeability could be altered by serum from preeclampsia (PE). METHODS: Confluent normal endothelial cells (ECs) were incubated with 20% serum from nonpregnant females, normal and PE pregnancies or combined with antioxidant superoxide dismutase (SOD) for 8 hr. Confluent PE ECs were incubated with 20% serum from normal pregnancies. EC barrier function of monolayer permeability was accessed by measuring EC electrical resistance (ER) and the leakage of horseradish peroxidase (HRP) passing through EC filters. Plasma concentrations of IL-8 and lipid peroxides by MDA were also measured. We determined 1) if serum from PE could affect EC permeable function; 2) if antioxidant and serum from normal pregnancies could preserve PE EC barrier function; 3) if lipid peroxides and cytokine IL-8 were increased in PE blood samples. Data are presented as mean+/-SE. ANOVA was used for statistical analysis. A p level less than 0.05 was considered statistically different. RESULTS: 1) ER was significantly decreased and HRP passage was significantly increased in ECs incubated with serum from PE compared to serum from non-pregnant and normal pregnant females (ER: 36.30+/-2.60 vs. 51.30+/-4.00 and 53.90+/-5.80 Omega x cm2, p<0.01; HRP: 0.100+/-0.020 vs. 0.014+/-0.002 and 0.022+/-0.007 DeltaOD470 nm, p<0.01, respectively). 2) ER was improved in PE ECs incubated with serum from normal pregnancies compared to controls, 52.28+/-3.13 vs. 34.50+/-3.80 Omega x cm2, p<0.01. 3) SOD attenuated decreased EC ER induced by PE serum, 55.58+/-3.61 Omega x cm2 (SOD+PE serum) vs. 42.34+/-3.24 (control) and 35.46+/ 2.44 (PE serum), p<0.01, respectively. 4) Both MDA and IL-8 concentrations were higher in plasma or serum samples from PE than those in samples from nonpregnancies and normal pregnancies, MDA: 28.65+/-1.45 vs. 22.40+/-1.47 and 25.53+/-0.89 micromol/mL, p<0.01; IL-8: 5.35+/-1.08 vs. 1.69+/-0.47 and 2.28+/ 0.73 pg/mL, p<0.05, respectively. Conclusions. 1) Sera from PE but not from nonpregnant women or normal pregnancies increase EC monolayer permeability. 2) Increased lipid peroxides and IL-8 are candidates altering EC barrier function. 3) Antioxidant SOD preserves increased EC monolayer permeability induced by PE serum, suggesting that EC oxidative stress may be associated with altered EC barrier function in preeclampsia. PMID- 12648449 TI - The calculus of immunity: quantitating antigen processing. AB - Peptide ligands destined for binding MHC class I molecules are generated by the proteasome in the cytoplasm and transported to the endoplasmic reticulum for loading onto newly synthesized class I molecules. A new study now provides a quantitative analysis of how total protein turnover in the cell relates to the efficiency of generating peptide epitopes presented by class I molecules. PMID- 12648450 TI - Caspase activation by granzyme B is indirect, and caspase autoprocessing requires the release of proapoptotic mitochondrial factors. AB - Apoptosis in response to granzyme B involves activation of caspase-dependent target cell death pathways. Herein, we show that granzyme B initiates caspase processing but cannot fully process procaspase-3 in intact Jurkat T leukemia or NT2 neuronal cells. Rather, the release from mitochondria of proapoptotic mediators cytochrome c, Smac/Diablo, and HtrA2/Omi facilitates full activation of caspases that results from autoprocessing. Bcl-2 overexpression in mitochondria suppresses the release of these proapoptotic molecules, resulting in cell survival despite partial procaspase processing by granzyme B. We propose that binding of inhibitor of apoptosis (IAP) proteins to partially processed procaspases inhibits cell death unless mitochondrial disruption also occurs in response to granzyme B or activated BH3-domain proteins such as truncated Bid. PMID- 12648451 TI - The survival of antigen-stimulated T cells requires NFkappaB-mediated inhibition of p73 expression. AB - We have explored the interactions between the NFkappaB and Cdk-Rb-E2F pathways in controlling T cell fate following antigen stimulation. The inhibition of NFkappaB in antigen-stimulated T cells results in apoptosis but does not inhibit E2F activation and S phase entry. IkappaB-induced apoptosis coincides with the superinduction of p73 expression and activity. G1 Cdk activity is required for IkappaB-induced apoptosis and the induction of p73. Importantly, p73 deficiency rescues activated T cells from the apoptosis resulting from the inhibition of NFkappaB. Thus, Cdk2 activation sends signals for both cell cycle progression and apoptosis, the latter of which must be blocked by NFkappaB to allow for proliferation. PMID- 12648452 TI - Quantitating protein synthesis, degradation, and endogenous antigen processing. AB - Using L929 cells, we quantitated the macroeconomics of protein synthesis and degradation and the microeconomics of producing MHC class I associated peptides from viral translation products. To maintain a content of 2.6 x 10(9) proteins, each cell's 6 x 10(6) ribosomes produce 4 x 10(6) proteins min(-1). Each of the cell's 8 x 10(5) proteasomes degrades 2.5 substrates min(-1), creating one MHC class I-peptide complex for each 500-3000 viral translation products degraded. The efficiency of complex formation is similar in dendritic cells and macrophages, which play a critical role in activating T cells in vivo. Proteasomes create antigenic peptides at different efficiencies from two distinct substrate pools: rapidly degraded newly synthesized proteins that clearly represent defective ribosomal products (DRiPs) and a less rapidly degraded pool in which DRiPs may also predominate. PMID- 12648453 TI - Granzyme B-induced apoptosis requires both direct caspase activation and relief of caspase inhibition. AB - Cytotoxic lymphocytes employ Granzyme B as a potent initiator of apoptosis to cleave and activate effector caspases. Unexpectedly, cells transfected with Bcl-2 were resistant to granzyme B-induced killing, suggesting that a mitochondrial pathway was critical. Utilizing cells expressing a dominant-negative caspase 9, the current study demonstrated that caspase activation via the apoptosome was not required. Indeed, cleavage of caspase 3 to p20 still occurred in Bcl-2 transfectants but processing to p17 was blocked. This blockade was recapitulated by the Inhibitor-of-Apoptosis-Protein XIAP and relieved by Smac/DIABLO. Thus granzyme B mediates direct cleavage of caspase 3 and also activates mitochondrial disruption, resulting in the release of proapoptotic proteins that suppress caspase inhibition. Engagement of both pathways is critical for granzyme-induced killing. PMID- 12648454 TI - Regulatory dendritic cells protect mice from murine acute graft-versus-host disease and leukemia relapse. AB - We have established a novel immunotherapeutic approach involving dendritic cells (DCs) with potent immunoregulatory property (designated as regulatory DCs [rDCs]) for acute graft-versus-host disease (GVHD) and leukemia relapse in allogeneic bone marrow (BM) transplantation (BMT) in mice bearing leukemia. rDCs displayed high levels of MHC molecules and extremely low levels of costimulatory molecules. A single injection of rDCs following allogeneic BMT controlled the ability of the transplanted T cells to induce acute GVHD and graft-versus-leukemia (GVL) effect in the recipients bearing leukemia, and that resulted in protection from the lethality caused by acute GVHD and tumor burden. Thus, the use of rDCs may be therapeutically useful for the treatment of acute GVHD and leukemia relapse in allogeneic BMT. PMID- 12648455 TI - Perforin is required for innate and adaptive immunity induced by heat shock protein gp96. AB - Tumor-secreted gp96-Ig is highly immunogenic and triggers CD8 T cell-mediated tumor rejection. In vivo secreted gp96-Ig and gp96-myc cause NK activation and clonal expansion of specific CD8(+) CTL in wild-type and in Fas-ligand-deficient (gld) mice but not in perforin- (PKO) or IFN-gamma-deficient (GKO) mice. Transfer of perforin-competent NK cells restores the ability of PKO mice to clonally expand CD8 CTL in response to gp96-Ig. The data demonstrate an essential role for perforin-mediated functions in the activation of innate and adaptive immunity by heat shock protein gp96-peptide complexes. Crosspresentation of antigens by heat shock proteins seems to require a perforin-dependent positive feedback loop between NK and DC for both sustained NK activation and clonal CTL expansion. The studies also explain how depressed NK activity in patients with tumors or after viral infections could diminish CTL responses. PMID- 12648456 TI - Regulation of murine cerebral malaria pathogenesis by CD1d-restricted NKT cells and the natural killer complex. AB - NKT cells are specialized cells coexpressing NK and T cell receptors. Upon activation they rapidly produce high levels of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) and are therefore postulated to influence T(H)1/T(H)2 immune responses. The precise role of the CD1/NKT cell pathway in immune response to infection remains unclear. We show here that CD1d-restricted NKT cells from distinct genetic backgrounds differentially influence T(H)1/T(H)2 polarization, proinflammatory cytokine levels, pathogenesis, and fatality in the P. berghei ANKA/rodent model of cerebral malaria. The functional properties of CD1d restricted NKT cells vary according to expression of loci of the natural killer complex (NKC) located on mouse chromosome 6, which is shown here to be a significant genetic determinant of murine malarial fatalities. PMID- 12648457 TI - SWAP-70-like adapter of T cells, an adapter protein that regulates early TCR initiated signaling in Th2 lineage cells. AB - We describe the isolation of a protein, SWAP-70-like adapter of T cells (SLAT), which is expressed at high levels in thymocytes and differentiated Th2 cells. SLAT expression was upregulated in differentiating Th2 cells and downregulated in Th1 cells. Ectopic SLAT expression exerted positive or negative effects on IL-4 versus IFNgamma induction, respectively. TCR signaling induced translocation of SLAT to the immunological synapse and its association with ZAP-70 kinase. SLAT reduced the association of ZAP-70 with TCR-zeta and interfered with ZAP-70 but not Lck signaling. Consistent with these results, pharmacological inhibition of ZAP-70 also induced Th2 skewing. Thus, SLAT is a protein which plays a role in Th2 development and/or activation, perhaps by interfering with ZAP-70 signaling. PMID- 12648458 TI - GATA-3 suppresses Th1 development by downregulation of Stat4 and not through effects on IL-12Rbeta2 chain or T-bet. AB - To further understand the interaction among GATA-3, Stat4, and T-bet in helper T cell development, we first showed that retroviral expression of GATA-3 in developing Th1 cells suppresses Th1 development through downregulation of Stat4 rather through downregulation of the IL-12Rbeta2 chain. Correspondingly, Stat4 levels are greatly suppressed during physiological Th2 development. Then, using cells doubly infected with GFP- and YFP-expressing retroviruses, we showed that retroviral GATA-3 expression in developing Th1 cells does not block Th1 development in cells coexpressing Stat4 but does so in cells coexpressing T-bet. Finally, we showed that retroviral Stat4 expression could facilitate Th2-->Th1 conversion in cells bearing an IL-12Rbeta2 transgene, even in cells lacking T bet. These findings reassert that Stat4 signaling is a central element of Th1/Th2 development. PMID- 12648459 TI - The cytosolic endopeptidase, thimet oligopeptidase, destroys antigenic peptides and limits the extent of MHC class I antigen presentation. AB - Most antigenic peptides presented on MHC class I molecules are generated by proteasomes during protein breakdown. It is unknown whether these peptides are protected from destruction by cytosolic peptidases. In cytosolic extracts, most antigenic peptides are degraded by the metalloendopeptidase, thimet oligopeptidase (TOP). We therefore examined whether TOP destroys antigenic peptides in vivo. When TOP was overexpressed in cells, class I presentation of antigenic peptides was reduced. In contrast, TOP overexpression didn't reduce presentation of peptides generated in the endoplasmic reticulum or endosomes. Conversely, preventing TOP expression with siRNA enhanced presentation of antigenic peptides. TOP therefore plays an important role in vivo in degrading peptides released by proteasomes and is a significant factor limiting the extent of antigen presentation. PMID- 12648460 TI - Nonpathogenic SIV infection of sooty mangabeys is characterized by limited bystander immunopathology despite chronic high-level viremia. AB - HIV-infected humans and SIV-infected rhesus macaques who remain healthy despite long-term infection exhibit exceptionally low levels of virus replication and active antiviral cellular immune responses. In contrast, sooty mangabey monkeys that represent natural hosts for SIV infection do not develop AIDS despite high levels of virus replication and limited antiviral CD8(+) T cell responses. We report here that SIV-infected mangabeys maintain preserved T lymphocyte populations and regenerative capacity and manifest far lower levels of aberrant immune activation and apoptosis than are seen in pathogenic SIV and HIV infections. These data suggest that direct consequences of virus replication alone cannot account for progressive CD4(+) T cell depletion leading to AIDS. Rather, attenuated immune activation enables SIV-infected mangabeys to avoid the bystander damage seen in pathogenic infections and protects them from developing AIDS. PMID- 12648462 TI - EGF receptor ligands. PMID- 12648463 TI - Neuregulins: functions, forms, and signaling strategies. AB - The neuregulins (NRGs) are cell-cell signaling proteins that are ligands for receptor tyrosine kinases of the ErbB family. The neuregulin family of genes has four members: NRG1, NRG2, NRG3, and NRG4. Relatively little is known about the biological functions of the NRG2, 3, and 4 proteins, and they are considered in this review only briefly. The NRG1 proteins play essential roles in the nervous system, heart, and breast. There is also evidence for involvement of NRG signaling in the development and function of several other organ systems, and in human disease, including the pathogenesis of schizophrenia and breast cancer. There are many NRG1 isoforms, raising the question "Why so many neuregulins?" Study of mice with targeted mutations ("knockout mice") has demonstrated that isoforms differing in their N-terminal region or in their epidermal growth factor (EGF)-like domain differ in their in vivo functions. These differences in function might arise because of differences in expression pattern or might reflect differences in intrinsic biological characteristics. While differences in expression pattern certainly contribute to the observed differences in in vivo functions, there are also marked differences in intrinsic characteristics that may tailor isoforms for specific signaling requirements, a theme that will be emphasized in this review. PMID- 12648464 TI - Epidermal growth factor receptor: mechanisms of activation and signalling. AB - The epidermal growth factor (EGF) receptor (EGFR) is one of four homologous transmembrane proteins that mediate the actions of a family of growth factors including EGF, transforming growth factor-alpha, and the neuregulins. We review the structure and function of the EGFR, from ligand binding to the initiation of intracellular signalling pathways that lead to changes in the biochemical state of the cell. The recent crystal structures of different domains from several members of the EGFR family have challenged our concepts of these processes. PMID- 12648465 TI - The deaf and the dumb: the biology of ErbB-2 and ErbB-3. AB - ErbB-2 (also called HER2/neu) and ErbB-3 are closely related to the epidermal growth factor receptor (EGFR/ErbB-1), but unlike EGFR, ErbB-2 is a ligandless receptor, whereas ErbB-3 lacks tyrosine kinase activity. Hence, both ErbB-2 and ErbB-3 are active only in the context of ErbB heterodimers, and ErbB-2. ErbB-3 heterodimers, which are driven by neuregulin ligands, are the most prevalent and potent complexes. These stringently controlled heterodimers are repeatedly employed throughout embryonic development and dictate the establishment of several cell lineages through mesenchyme-epithelial inductive processes and the interactions of neurons with muscle, glia, and Schwann cells. Likewise, the potent combination of signaling pathways engaged by the heterodimers drives an aggressive phenotype of tumors of secretory epithelia, including breast and lung cancers. This review highlights recent structural insights into the mechanism of ligand-induced heterodimer formation, and concentrates on signaling pathways employed by ErbB-2 and ErbB-3 in normal and in malignant cells. PMID- 12648466 TI - ErbB-4: mechanism of action and biology. AB - The most recently described member of the ErbB receptor tyrosine kinase family is ErbB-4. In general, the structure of this receptor and its mechanism of action is similar to that described for ErbB-1. However, significantly less is known about ErbB-4 and there are several novel aspects to its structure, mechanism of action, and biology. This includes the spectrum of ligands that activate ErbB-4, the presence of functionally distinct isoforms, a proteolytic processing pathway to the nucleus, and the capacity to induce a spectrum of cellular responses such as mitogenesis, differentiation, growth inhibition, and survival. PMID- 12648467 TI - Trafficking of the ErbB receptors and its influence on signaling. AB - Although members of the ErbB receptor family are found predominantly at the cell surface, these receptors undergo constant cycling between the plasma membrane and the endosomal compartment. In the absence of an activating ligand, these receptors are slowly internalized (t(1/2) approximately 30 min) but are quickly recycled. The constitutive degradation rate of the epidermal growth factor (EGF) receptor (EGFR) is slower than other ErbB family members and only the EGFR appears to alter its trafficking pattern in response to ligand binding. This altered pattern is characterized by accelerated internalization and enhanced lysosomal targeting. Ligand-regulated trafficking of the EGFR is mediated by a series of motifs distributed through the cytoplasmic domain of the receptor that are exposed by a combination of activation-mediated conformation changes and the binding of proteins such as Grb2. As a consequence of induced internalization, most EGFR signaling occurs within endosomes whereas signaling by the other members of the ErbB family appear to be generated predominantly from the cell surface. Overexpression of ErbB family members can disrupt normal receptor trafficking by driving heterodimerization of receptors with disparate trafficking patterns. Because different ErbB receptor substrates are localized in different cellular compartments, disrupted trafficking could be an important factor in the altered signaling patterns observed as a consequence of receptor overexpression. PMID- 12648468 TI - ErbBs in mammary development. AB - Members of the ErbB (epidermal growth factor receptor) family of receptor tyrosine kinases are important in etiology of human mammary carcinoma, and are rational targets for cancer therapy. The frequent selection of ErbB2/HER2/Neu, and, less often, the epidermal growth factor receptor, and ErbB3 for overexpression in breast cancer implies that these receptors have important functions in normal mammary development. Better understanding of ErbBs in mammary development may yield important dividends for development and deployment of cancer therapies. The roles of these receptors and their ligands in mammary development are discussed, with an emphasis on new insights from genetic analysis of the receptors in mice. PMID- 12648469 TI - The ErbB receptors and their role in cancer progression. AB - The involvement of the ErbB receptor tyrosine kinases in human cancer, as well as their essential role in a variety of physiological events during normal development, have motivated the interest in this receptor family. Approaches taken to block the activity of ErbB receptors in cancer cells have not only proven that they drive in vitro tumor cell proliferation, but have also become clinically relevant for targeting tumors with deregulated ErbB signaling. The mechanisms and downstream effectors through which the ErbB receptors influence processes linked to malignant development, including proliferation, cell survival, angiogenesis, migration, and invasion, are, however, only now becoming apparent. Our particular emphasis in this review will be on how ErbB receptors, in particular ErbB1 and ErbB2, contribute to processes linked to cancer progression. Importantly, in keeping with the emerging theme that ErbB receptors do not function in isolation, we will focus on receptor cooperativity, i.e., ErbB1 cooperates with other classes of receptors, and the ligand-less ErbB2 functions as a heterodimer with other ErbBs. PMID- 12648470 TI - Ligand-independent oncogenic signaling by the epidermal growth factor receptor: v ErbB as a paradigm. AB - Relay of information from the extracellular environment into the cell often results from a peptide growth factor binding to its cognate cell surface receptor; this event is an integral mechanism by which many cellular functions occur, including cell growth, motility, and survival. In recent years, however, this requirement for ligand binding has been shown to be surpassed by several distinct mechanisms, including cell surface receptor cross-talk (e.g., between epidermal growth factor receptor [EGFR] and G-coupled receptors), receptor extracellular matrix interactions (e.g., EGFR: integrin complexes), and finally by structural mutations within the receptor itself. While all of these pathways result in so-called ligand-independent signaling by the EGF receptor, to date, only structural mutations in the receptor have been shown to result in qualitative changes in downstream targets of the receptor, which specifically result in oncogenic signaling, transformation, and tumorigenicity. In this review, we describe aspects of the known signaling properties of the retroviral oncogene v-ErbB as a model of ligand-independent oncogenic signaling, and compare these properties to results emerging from ongoing studies on structurally related EGF receptor mutants originally identified in human tumors. A better understanding of the signaling pathways used by these uniquely oncogenic receptor tyrosine kinase mutants may ultimately reveal new targets for the development of novel therapeutics selective for the inhibition of tumor cell growth. PMID- 12648471 TI - ErbB-targeted therapeutic approaches in human cancer. AB - The overexpression and aberrant function of the epidermal growth factor receptor (EGFR, erbB1, HER1) and its ligands and coreceptors in a wide spectrum of epithelial cancers have provided a rationale for targeting this signaling network with novel treatment approaches. Several antireceptor therapeutic strategies have been pursued, but two stand ahead in their clinical development. One approach has been the generation of small molecules that compete with adenosine triphosphate (ATP) for binding to the receptor's kinase pocket, thus blocking receptor activation and the transduction of postreceptor signals. The second approach utilizes humanized monoclonal antibodies generated against the receptor's ligand binding extracellular domain. These antibodies block binding of receptor activating ligands and, in some cases, can induce receptor endocytosis and downregulation. Clinical studies already suggest that both of these approaches, either alone or in combination with standard anticancer therapies, are well tolerated and can induce clinical responses and tumor stabilization in a variety of common carcinomas. PMID- 12648472 TI - Mechanism of action of erbB tyrosine kinase inhibitors. AB - Over the last decade, drug discovery efforts have generated a myriad of compounds that inhibit the activity of the erbB family of tyrosine kinases with potencies and selectivity that have surpassed original expectations. These characteristics, along with improved pharmaceutical properties, have enabled inhibitors from this class of agents to finally realize their therapeutic potential, and indeed, some are currently producing significant clinical responses. Interestingly, those properties that are essential for a clinically active inhibitor of the erbB family are most readily attained with compounds that bind at the ATP site, and the most successful compounds have shown a distinct convergence to certain common chemical features. The reasons for this trend are beginning to be realized through the generation of an increasing array of crystalline structures for protein kinases as well as advances in molecular modeling. This has allowed a more complete understanding of the precise physical interactions that occur between erbB tyrosine kinase inhibitors and their target(s), which, in turn, has begun to shed light on the mechanism by which these molecules attain their remarkable affinity and specificity. PMID- 12648474 TI - The epidermal growth factor system in Caenorhabditis elegans. AB - The single known epidermal growth factor-like growth factor and single epidermal growth factor receptor in Caenorhabditis elegans mediate two types of processes, each via a distinct signal transduction pathway. Several instances of cell fate specification during organogenesis require the RAS-MAP kinase pathway, as well as multiple nuclear factors. By contrast, appropriate myoepithelial contractions during ovulation involve IP3-mediated signal transduction. Positive modulators of the RAS pathway include KSR, SUR-8, phosphatase PP2A, and a zinc cation diffusion facilitator. Negative regulators of the RAS pathway include homologs of CBL, GAP 1, ACK, and MAP kinase phosphatase, while negative regulators of the IP3 pathway are enzymes that modify IP3. In addition to its stimulation of RAS activity, the GRB2 homolog SEM-5 acts negatively on both signaling pathways, as does the Ack related kinase ARK-1. PMID- 12648473 TI - Signaling by the Drosophila epidermal growth factor receptor pathway during development. AB - In 1997 we wrote a review entitled "A thousand and one roles for the Drosophila epidermal growth factor (EGF) receptor (DER/EGFR)." We are not there yet in terms of the number of developmental roles assigned to this receptor in Drosophila. Nevertheless, DER has certainly emerged as one of the key players in development, since it is used repeatedly to direct cell fate choices, cell division, cell survival, and migration. A battery of activating ligands and an inhibitory ligand achieves this versatility. For the ligands that are produced as membrane-bound precursors, trafficking and processing are the key regulatory steps, determining the eventual temporal and spatial pattern of receptor activation. In most cases DER is activated at a short range, in the cells adjacent to the ones producing the active ligand. This activation dictates a binary choice. In some instances DER is also activated over a longer range, and multiple cell fate choices may be induced, according to its level of activation. A battery of negative feedback loops assures the limited range of DER induction. The distinct responses to DER activation in the different tissues depend upon combinatorial interactions with other signaling pathways and tissue-specific factors, at the level of target-gene regulation. PMID- 12648475 TI - Lack of correlation between environmental or biological indicators of benzene exposure at parts per billion levels and micronuclei induction. AB - Despite growing concern for possible carcinogenic effects associated with environmental benzene exposure in the general population, few studies exist at parts per billion (ppb) levels. We investigated the existence of a relationship between airborne/biological measurements of benzene exposure (i.e., personal/area sampling and unmodified urinary benzene/trans,trans-muconic acid; t,t-MA) and micronuclei induction (cytochalasin B technique) among exposed chemical laboratory workers (n=47) and traffic wardens (n=15). Although urinary t,t-MA (106.9+/-123.17 microg/L(urine)) correlated (R(2)=0.37) with urinary benzene (0.66+/-0.99 microg/L(urine)), neither biological measurement correlated with environmental benzene exposure (14.04+/-9.71 microg/m(3); 4.39+/-3.03ppb), suggesting that, at ppb level (1ppb=3.2 microg/m(3)), airborne benzene constitutes a fraction of the total intake. Traffic wardens and laboratory workers had comparable numbers of micronuclei (4.70+/-2.63 versus 5.76+/-3.11; n.s.), similar to levels recorded in the general population. With univariate/multivariate analysis, no association was found between micronuclei induction and air/urinary benzene exposure variables. Notably, among the personal characteristics examined (including age, gender, smoking, drinking, etc.), high body mass index correlated with micronuclei induction while, among females, use of hormonal medication was associated with less micronuclei. Thus the present study provides no evidence that ppb levels of environmental benzene exposure appreciably affect micronuclei incidence (against the background of other relevant factors). However, this should not be taken as an argument against efforts aiming to reduce environmental benzene pollution. PMID- 12648476 TI - Dietary and reproductive determinants of plasma organochlorine levels in pregnant women in Rio de Janeiro. AB - Organochlorine (OC) plasma levels and their dietary and reproductive determinants were investigated in 64 pregnant women from Rio de Janeiro, Brazil. Prenatal exposure of newborns was evaluated in a subset of these women in which umbilical cord blood was analyzed. To assess the influence of dietary factors on OC levels, a semiquantitative food frequency questionnaire was completed by participants. Nondietary determinants were investigated through a general questionnaire that addressed reproductive history, lifestyle, and social-demographic characteristics. Both women and their newborns had detectable but low levels of OCs. The most frequently detected compound was p,p'-DDE, the main metabolite of p,p'-DDT. This compound was found in 97% of the women who participated in the study and in 67% of the umbilical cord blood samples, with levels ranging from 0.161 to 8.03ppb and from 0.320 to 1.06ppb, respectively. Among the OCs detected, only p,p'-DDE showed any correlation with dietary and nondietary factors. There was a positive correlation between maternal p,p'-DDE and the consumption of fish (Pearson r=0.38,P=0.002) and chicken (Pearson r=0.26, P=0.042). The correlation between pork consumption and p,p'-DDE exhibited an inverse relationship (Pearson r=-0.25,P=0.052). Parity was the only nondietary factor that showed a significant correlation (Pearson r=-0.36,P=0.004). The main determinants of p,p'-DDE levels in this group of women were fish consumption and parity, together explaining 28% of the variance (P<001), in a multivariate model. PMID- 12648477 TI - Respiratory impairment due to asbestos exposure in brake-lining workers. AB - There is extensive evidence that exposure to asbestos causes pulmonary parenchymal fibrosis, pleural disease, and malignant neoplasm in asbestos-exposed workers. However, few data concerning brake-lining workers are available in the literature. In this study, we aimed to assess the long-term effects of chrysotile asbestos exposure on lung function and the risk of asbestos-related diseases in brake-lining workers. Seventy-four asbestos-exposed workers who processed brake lining products and 12 unexposed office workers were offered pulmonary function tests (spirometry and transfer factor) in 1992 and 1999. In 1999, the mean duration of asbestos exposure was 10.00+/-4.07 and 11.02+/-4.81 years (7-31 years) in nonsmoking and smoking asbestos workers, respectively. Transfer factor (T(L), CO) and transfer coefficient (K(CO)) decline were significant in the 7 year follow-up in both smoking and nonsmoking asbestos workers. However, lung function indices of the control group, whom were all current smokers; were also found to be decreased, including FEF(75), T(L), CO and K(CO). We found minimal reticular changes in 10 asbestos workers who were all current smokers, they underwent high-resolution computed tomography scans of the chest and we found that they had peribronchial thickening resulting from smoking. As a conclusion, even in the absence of radiographic asbestosis, T(L), CO and K(CO) may decrease after a mean 10-year duration of exposure to asbestos in brake-lining workers and this is more noticeable with cigarette burden. PMID- 12648478 TI - Effects of endosulfan on brain acetylcholinesterase activity in juvenile bluegill sunfish. AB - The effects of endosulfan upon brain acetylcholinesterase (AChE) activity were measured in juvenile bluegill sunfish (Lepomis macrochirus). Based on exposure durations of 0, 24, 48, 72, and 96h and 1 week at 1.0 microg/L (just below the LC50 of 1.2 microg/L for this species), step-wise decreases in AChE activity were noted, corresponding to 0%, 3.57%, 12.65%, 14.23%, 16.31%, and 23.11% inhibition, respectively. Total brain protein concentrations were measured to test the accuracy of the AChE data with no significant anomalies. The duration of exposure was related to the reduction in the AChE activities which reflected the biotoxicity of endosulfan. The changes in the AChE activities will certainly affect the normal behavior of the juvenile bluegill which is detrimental to their very existence in the natural habitat. PMID- 12648479 TI - Public views on the links between air pollution and health in Northeast England. AB - We investigated how public perceptions of the links between air pollution and health varied with contextual factors describing individuals and their locality. Information was collected via postal surveys on 2744 adults resident in five neighborhoods in Northeast England. Perceptions were compared by individual factors (health status, age, and gender) and locality factors (relative deprivation, proximity to industry and district-Teesside or Sunderland, with different amounts of heavy industry). There was relatively little variation in views about air pollution and health links between neighborhoods. The greatest contrasts were found when comparing those living near or further from industry and between the two districts. Any differences were related more to awareness of illness in the neighborhood thought to be affected by air pollution, rather than belief that a particular disease was linked to air pollution. Chronic illness status and age were sometimes found to be associated with perceptions of disease affected by air pollution, but gender and material deprivation were not central to differences in risk perceptions among the population studied. In understanding public perceptions about the links between air quality and health, research should focus on the characteristics of places as well as of people. PMID- 12648480 TI - Associations between dietary intake and serum polychlorinated dibenzo-p-dioxin and dibenzofuran (PCDD/F) levels in Taiwanese. AB - The study was conducted to correlate the consumption frequency of different food groups (milk, eggs, fish, and others) and the levels of serum polychlorinated dibenzo-p-dioxin and dibenzofuran (PCDD/F) concentrations of residents living near a municipal waste incinerator. All selected subjects were between 18 and 65 years old and proportionally chosen from each age group based on population distribution. In addition, they had to have lived within a 5-km radius of the incinerator for at least 5 years. Trained interviewers administered a dietary questionnaire interview. Higher serum PCDD/F levels were found in older than in younger subjects, and higher PCDD/F concentrations were found in females than in males. An analysis between the consumption frequency of different foods and serum PCDD/F levels showed that fish might have contributed the highest quantity of measured serum PCDD/Fs. Yet, the regression coefficient of dietary intake and PCDD/F concentration was only 0.017 before and 0.105 after adjusting for the variables of age, gender, and smoking status of the study subjects. The frequency of fish consumption may be the most significant contributor to serum PCDD/F levels. Further research is needed to quantify the association between the consumption of various food groups and their potential contributions to the corresponding serum PCDD/F concentrations. PMID- 12648481 TI - Evaluation of 4-nonylphenol toxicity in the clam Tapes philippinarum. AB - Lethal and sublethal effects of 4-nonylphenol (NP) were investigated in the clam Tapes philippinarum from the Lagoon of Venice. In a 96-h lethality test, bivalves were exposed to the following NP concentrations: 0, 0+acetone, 0.19, 0.38, 0.75, 1.5, and 3.0mg NP/L. The 96-h LC(50) value was 1.12mg NP/L. No mortality was observed at 0.19mg NP/L, whereas at 1.5 and 3.0mg NP/L both siphons and foot of clams were often cut-off, the animals being unable to withdraw them before shell closure, suggesting the possible narcotic effect of NP. Lower concentrations (0, 0+acetone, 0.025, 0.05, 0.1, and 0.2mg NP/L) were then used to evaluate sublethal NP effects on clearance rate (CR), respiration rate (RR), scope for growth (SFG), and survival in air. Following a 7-day exposure, decreased RRs were found at all NP concentrations tested. Significant decreases in the CR and SFG were only observed at the two highest NP concentrations (0.1 and 0.2mg/L), with SFG reductions reaching 54% and 71%, respectively. Last, the highest concentrations tested significantly decreased the resistance of clams to exposure to air and enhanced the mortality rate. As the lowest effective NP concentrations are similar to environmentally realistic levels, a condition of potential risk for the well-being of clam populations in estuarine areas is highlighted. PMID- 12648482 TI - Lichens as biomonitors around a coal-fired power station in Israel. AB - In the present study epiphytic lichens were applied as biomonitors of air pollution to determine the environmental impact of a coal-fired power station. Thalli of the lichen Ramalina lacera (With.) J.R. Laund. growing on carob twigs (Ceratonia siliqua L.) were collected with their substrate in July 2000 in a relatively unpolluted forest near HaZorea, Ramoth Menashe, Northeast Israel, and transplanted to 10 biomonitoring sites in the vicinity of the coal-fired power station Oroth Rabin near the town of Hadera. The lichens were retrieved in January 2001. We examined the following parameters of lichen vitality: (a) potential quantum yield of photosynthesis expressed as fluorescence ratio F(v)/F(m), (b) stress-ethylene production, and (c) electric conductivity expressing integrity of cell membranes. Following an exposure of 7 months, the lichens were retrieved and physiological parameters and data of elemental content were analyzed comparatively. Electric conductivity values correlated positively with B, Fe, Mg, Mn, Na, Pb, S, Sn, and Ti content. Concentrations of stress ethylene correlated positively with Al, Ba, Pb, S, and V content and negatively with Cu and Sn. F(v)/F(m) ratios correlated negatively with S content. Some of the heavy metals reached lower levels than those reported in the relevant literature despite a wind regime that should have blown pollutants toward the biomonitoring sites. PMID- 12648483 TI - fc177, a minor dec-1 proprotein, is necessary to prevent ectopic aggregation of the endochorion during eggshell assembly in Drosophila. AB - The Drosophila eggshell is a highly specialized extracellular matrix that forms between the oocyte and the surrounding epithelial follicle cells during late oogenesis. The dec-1 gene, which is required for proper eggshell assembly, produces three proproteins that are cleaved within the vitelline membrane layer to multiple derivatives. The different spatial distributions of the cleaved derivatives suggest that they play distinct roles in eggshell assembly. Using extant dec-1 mutations in conjunction with genetically engineered dec-1 transgenes, we show that, although all three dec-1 proproteins, fc106, fc125, and fc177, are required for female fertility, gross morphological abnormalities in the eggshell are observed only in the absence of fc177. The coalescence of the roof, pillar, and floor substructures of the tripartite endochorion suggested that quantitatively minor fc177 derivatives are necessary to prevent ectopic aggregation of endochorion proteins during the assembly process. Expression of a fc177 cDNA in dec-1 null mutants was sufficient to restore spaces within the endochorion layer. Fc177 may function as a scaffolding protein akin to those utilized in viral morphogenesis. PMID- 12648484 TI - Sex-specific telomere redistribution and synapsis initiation in cattle oogenesis. AB - The process of homolog pairing is well characterised in meiosis of male mammals, but much less information is available from female meiosis. We have therefore studied telomere dynamics by FISH and synapsis formation by immunostaining of synaptonemal complex proteins (SCP3, SCP1) on ovarian sections from 15 bovine fetuses, which covered the entire female prophase I. Telomeres displayed a dispersed intranuclear distribution in oogonia and relocated to the nuclear periphery during the preleptotene stage. Tight telomere clustering (bouquet formation) coincided with synapsis initiation at the leptotene/zygotene transition. Clustering of telomeres persisted during zygotene and even into the pachytene stage in a subset of nuclei, while it was absent in diplotene/dictyotene stage nuclei. Thus, the bouquet stage in the bovine female lasts significantly longer than in the male. Further, we observed that synapsis in the female initiated both terminally and interstitially in earliest zygotene stage oocytes, which contrasts with the predominantly terminal synapsis initiation in early zygotene spermatocytes of the bovine male. Altogether, our data disclose a sex-specific difference in telomere dynamics and synapsis initiation patterns in male and female bovine germ cells that may be related to the sex-specific differences in recombination rates observed in this and other mammalian species. PMID- 12648485 TI - Evolutionary aspects of developmentally regulated helix-loop-helix transcription factors in striated muscle of jellyfish. AB - The function of basic helix-loop-helix (bHLH) proteins in cell differentiation was shown to be conserved from Drosophila to vertebrates, exemplified by the function of MyoD in striated muscle differentiation. In phylogeny striated muscle tissue appears first in jellyfish and the question of its evolutionary position is controversially discussed. For this reason we have studied the developmental role of myogenic bHLH genes in medusa development. Based on their dimerization ability, four genes of the bHLH family of transcription factors were isolated from the hydrozoan jellyfish Podocoryne carnea. While the proteins Id and Ash group with cognate family members from bilaterians, Net-like and JellyD1 could not be unequivocally classified. Id is expressed during the medusa budding process and in the adult medusa, Ash and Net-like are expressed in all life cycle stages from egg to adult medusa and JellyD1 is expressed in the blastula and gastrula stages, the planula larva, and in late medusa bud stages. The dimerization specificity, the expression pattern, and the conservation of two residues specific for a MyoD bHLH domain suggest that JellyD1 is related to an ancestral MyoD gene. Id, Net-like, and JellyD1 are either expressed in the entocodon or its derived tissues, the striated and smooth muscle of the bell. These findings strengthen the hypothesis that the entocodon is a mesoderm-like structure and that the common ancestor of Cnidaria and Bilateria was more complex in cell-type architecture and body organization than commonly thought. PMID- 12648486 TI - Galphaq and Galpha11 proteins mediate endothelin-1 signaling in neural crest derived pharyngeal arch mesenchyme. AB - Endothelin-A (ET(A)) is a G-protein-coupled receptor expressed in the neural crest-derived mesenchyme of the pharyngeal arches during craniofacial development. Targeted deletion of the ET(A) receptor or its ligand endothelin-1 (ET-1) causes cleft palate and hypoplasia of the mandible, otic cup, and tympanic ring. Previously we showed that Galpha(q)/Galpha(11)-null mice die around E11.0, whereas Galpha(q)((-/-))Galpha(11)((+/-)) mice survive to birth with hypomorphic phenotypes similar to, but less severe than, ET(A) or ET-1-null mice. To determine whether ET-1 signaling is transduced by Galpha(q)/Galpha(11) proteins, we examined the expression patterns of several ET-1 dependent and independent transcription factors in Galpha(q)/Galpha(11)-deficient embryos. Expression of genes encoding the ET-1-dependent transcription factors Dlx3, Dlx6, dHAND, and eHAND was specifically downregulated in the pharyngeal arches of Galpha(q)/Galpha(11)-deficient mice. In contrast, pharyngeal arch expression of the homeobox gene Msx1, which is not regulated by ET-1 signaling, was maintained in these embryos. We conclude that the Galpha(q) and Galpha(11) proteins serve as the intracellular mediators of ET-1 signaling in the pharyngeal arch mesenchyme. PMID- 12648487 TI - Hair cycle regulation of Hedgehog signal reception. AB - Proper patterning of self-renewing organs, like the hair follicle, requires exquisite regulation of growth signals. Sonic hedgehog (Shh) signaling in skin controls the growth and morphogenesis of hair follicle epithelium in part through regulating the Gli transcription factors. While ectopic induction of Shh target genes leads to hair follicle tumors, such as basal cell carcinomas, how Shh signaling normally functions during the cyclic process of hair development is unknown. Here, we show that, during the hair cycle, Shh expression and the ability of skin cells to respond to Shh signaling is spatially and temporally regulated. Induction of Shh target genes normally occurs only in the anagen hair follicle in response to expression of Shh. However, in patched1 heterozygous mice, putative tumor precursors form with concomitant induction of Shh target gene transcription only during anagen in follicular and interfollicular keratinocytes. Ectopic production of Gli1 accumulates Gli protein and induces Shh target genes and epithelial tumors at anagen but not other stages, pointing to a restricted competence occurring at the level of Gli protein accumulation. Delivery and reception of growth signals among multipotent cells are restricted in time and space to facilitate cyclic pattern formation. PMID- 12648488 TI - Requirement for RNA-binding activity of MSY2 for cytoplasmic localization and retention in mouse oocytes. AB - MSY2, a mouse germ cell-specific Y-box protein, is implicated in the global regulation of the stability and/or translation of maternal mRNAs in the mouse oocyte. We report here that in the oocyte approximately 75% of MSY2 protein is associated with a Triton-insoluble preparation, whereas in either male germ cells or when exogenously expressed in transfected somatic cells almost all MSY2 is soluble. This retention in the oocyte, which is unlikely mediated either by microfilaments or by microtubules, markedly decreases beyond the two-cell stage of development. By microinjecting mutant MSY2-EGFP chimeric mRNAs into mouse oocytes and then assaying the expressed protein's localization by laser-scanning confocal microscopy, we find that an intact cold-shock domain (CSD), containing two RNA-binding motifs, is required to localize MSY2 to the oocyte cytoplasm. In addition, an additional basic/aromatic amino acid island (B/A), which can also interact with RNA, in the C-terminal tail domain is necessary to retain MSY2 following Triton permeabilization. Intact mRNA appeared required for this retention, since RNase A treatment of Triton-permeabilized oocytes or microinjection of RNase A into the oocyte released essentially all of the endogenous MSY2 protein. Furthermore, there is a positive correlation between the ability of the mutant MSY2-EGFP protein to remain associated with the Triton insoluble preparations and its increased affinity for RNA, as determined by RNA electrophoretic mobility shift assays. These results suggest that binding of intact maternal mRNA by MSY2 is required for its cytoplasmic retention. PMID- 12648489 TI - Disruption of sonic hedgehog signaling alters growth and patterning of lingual taste papillae. AB - Taste buds on the anterior part of the tongue develop in conjunction with epithelial-mesenchymal specializations in the form of gustatory (taste) papillae. Sonic hedgehog (Shh) and Bone Morphogenetic Protein 4 (BMP4) are expressed in developing taste papillae, but the roles of these signaling molecules in specification of taste bud progenitors and in papillary morphogenesis are unclear. We show here that BMP4 is not expressed in the early tongue, but is precisely coexpressed with Shh in papillary placodes, which serve as a signaling center for both gustatory and papillary development. To elucidate the role of Shh, we used an in vitro model of mouse fungiform papillary development to determine the effects of two functional inhibitors of Shh signaling: anti-Shh (5E1) antibody and cyclopamine. Cultured E11.5 tongue explants express Shh and BMP4(LacZ) in a pattern similar to that of intact embryos, localizing to developing papillary placodes after 2 days in culture. Tongues cultured with 5E1 antibody continue to express these genes in papillary patterns but develop more papillae that are larger and closer together than in controls. Tongues cultured with cyclopamine have a dose-dependent expansion of Shh and BMP4(LacZ) expression domains. Both antibody-treated and cyclopamine-treated tongue explants also are smaller than controls. Taken together, these results suggest that, although Shh is not involved in the initial specification of papillary placodes, Shh does play two key roles during pmcry development: (1) as a morphogen that directs cells toward a nonpapillary fate, and (2) as a mitogen, causing expansion of the interplacodal epithelium and underlying mesenchyme. PMID- 12648490 TI - Mesenchymal expression of Foxl1, a winged helix transcriptional factor, regulates generation and maintenance of gut-associated lymphoid organs. AB - The Foxl1 gene, which encodes a winged helix transcriptional regulator, is expressed in the mesenchymal layer of developing and mature gastrointestinal tract. Foxl1-deficient mice exhibit various defects not only in the epithelial layer of the gastrointestinal tract but also in gut-associated lymphoid tissues. In the small intestine of Foxl1-deficient mice, the formation of Peyer's patches is affected, particularly in the caudal region. This alteration is shown to be due to the delayed formation of Peyer's patches organizing centers as revealed by the expressions of VCAM1 and IL-7 receptor alpha-chain at 17.5 days postcoitus. Peyer's patch defects are concordant with the significantly decreased expression of Lymphotoxin beta-receptor in the caudal region of fetal intestine. Foxl1 is suggested to regulate the responsiveness of fetal intestinal mesenchymal cells to inductive signals mediated by Lymphotoxins during Peyer's patch organogenesis. In addition, constitutive outgrowth of colonic patches due to defects in radioresistant stromal components of colonic patches are seen in Foxl1-deficient mice. Because of the functional similarities of hypertrophic colonic patches to those seen in hapten-induced experimental colitis, this hypertrophy is suggested to involve Lymphotoxin beta-receptor signaling. Together, the data suggest that Foxl1 might be involved in cellular responses of gut-associated lymphoid tissues dependent upon the Lymphotoxins/Lymphotoxin beta-receptor axis. PMID- 12648492 TI - The roles of Pax6 in the cornea, retina, and olfactory epithelium of the developing mouse embryo. AB - The roles of Pax6 were investigated in the murine eye and the olfactory epithelium by analysing gene expression and distribution of Pax6(-/-) cells in Pax6(+/+) <--> Pax6(-/-) chimeras. It was found that between embryonic days E10.5 and E16.5 Pax6 is autonomously required for cells to contribute fully not only to the corneal epithelium, where Pax6 is expressed at high levels, but also to the to the corneal stroma and endothelium, where the protein is detected at very low levels. Pax6(-/-) cells contributed only poorly to the neural retina, forming small clumps of cells that were normally restricted to the ganglion cell layer at E16.5. Pax6(-/-) cells in the retinal pigment epithelium could express Trp2, a component of the pigmentation pathway, at E14.5 and a small number went on to differentiate and produce pigment at E16.5. The segregation and near-exclusion of mutant cells from the nasal epithelium mirrored the behaviour of mutant cells in other developmental contexts, particularly the lens, suggesting that common primary defects may be responsible for diverse Pax6-related phenotypes. PMID- 12648491 TI - Difference in the maternal and zygotic contributions of tumorhead on embryogenesis. AB - Tumorhead (TH) is a maternally expressed gene in Xenopus laevis, that when overexpressed, increased proliferation of ectodermal derivatives and inhibited neural and epidermal differentiation. However, injection of anti-TH antibodies inhibited cleavage of all blastomeres, not only those contributing to the ectoderm. The injection of TH morpholino antisense oligonucleotide (TH-MO), which inhibits translation of TH mRNA, did not affect early cleavage but inhibited cell division in both the neural field and epidermis. This was accompanied by the inhibition of neural and epidermal markers. TH-MO did not affect the formation and differentiation of mesoderm and endoderm derivatives. Our overexpression and loss-of-function studies demonstrated that TH plays an important role in differentiation of the ectoderm by regulating cell proliferation. They also supported the conclusion that the maternal component of TH may affect the cell cycle in all cells, while the zygotic component has a germ layer-specific effect on the ectoderm. PMID- 12648494 TI - Erythropoietin and retinoic acid, secreted from the epicardium, are required for cardiac myocyte proliferation. AB - We have established a heart slice primary culture, which allows us to mechanically separate distinct cardiac cell populations and assay their relative mitogenic and trophic effects on cardiac myocyte proliferation and survival. Using this system, we have found that a signal(s) from the epicardium, but not the trabeculae and endocardium, is required in embryonic day 10 (E10) chick heart slices for continued cardiac myocyte proliferation and survival. An examination of potential epicardial growth or trophic factors has revealed that blockade of either retinoic acid (RA) or erythopoietin (epo) signaling from the epicardium inhibits cardiac myocyte proliferation and survival. The blockade of cardiac myocyte proliferation following administration of an RA antagonist can be rescued by exogenous epo. Conversely, the blockade of cardiac myocyte proliferation following administration of an anti-epo receptor antisera can be rescued by exogenous RA. Thus, our findings suggest that RA and epo signals work in parallel to support myocardial cell proliferation. In addition, we have found that these factors do not act directly on myocardial cells. Rather, they induce another soluble factor(s) in the epicardium that directly regulates proliferation of cardiac myocytes. We therefore postulate that the epicardium controls normal heart growth in ventricular segments of the embryonic chick heart by secreting a cardiac myocyte mitogen whose expression (or activity) is regulated by both RA and erythropoietin signaling. PMID- 12648493 TI - The tumour suppressor gene l(2)giant discs is required to restrict the activity of Notch to the dorsoventral boundary during Drosophila wing development. AB - During the development of the Drosophila wing, the activity of the Notch signalling pathway is required to establish and maintain the organizing activity at the dorsoventral boundary (D/V boundary). At early stages, the activity of the pathway is restricted to a small stripe straddling the D/V boundary, and the establishment of this activity domain requires the secreted molecule fringe (fng). The activity domain will be established symmetrically at each side of the boundary of Fng-expressing and non-expressing cells. Here, I present evidence that the Drosophila tumour-suppressor gene lethal (2) gaint discs (lgd) is required to restrict the activity of Notch to the D/V boundary. In the absence of lgd function, the activity of Notch expands from its initial domain at the D/V boundary. This expansion requires the presence of at least one of the Notch ligands, which can activate Notch more efficiently in the mutants. The results further suggest that Lgd appears to act as a general repressor of Notch activity, because it also affects vein, eye, and bristle development. PMID- 12648496 TI - Down-regulation of Notch target gene expression by Suppressor of deltex. AB - In Drosophila, Suppressor of deltex (Su(dx)) mutations display a wing vein gap phenotype resembling that of Notch gain of function alleles. The Su(dx) protein may therefore act as a negative regulator of Notch but its activity on actual Notch signalling levels has not been demonstrated. Here we show that Su(dx) does regulate the level of Notch signalling in vivo, upstream of Notch target genes and in different developmental contexts, including a previously unknown role in leg joint formation. Overexpression of Su(dx) was capable of blocking both the endogenous activity of Notch and the ectopic Notch signalling induced by the overexpression of Deltex, an intracellular Notch binding protein. In addition, using the conditional phenotype of the Su(dx)(sp) allele, we show that loss of Su(dx) activity is rapidly followed by an up-regulation of E(spl)mbeta expression, the immediate target of Notch signal activation during wing vein development. While Su(dx) adult wing vein phenotypes are quite mild, only affecting the distal tips of the veins, we show that the initial consequence of loss of Su(dx) activity is more severe than previously thought. Using a time course experiment we show that the phenotype is buffered by feedback regulation illustrating how signalling networks can make development robust to perturbation. PMID- 12648495 TI - Doubleridge, a mouse mutant with defective compaction of the apical ectodermal ridge and normal dorsal-ventral patterning of the limb. AB - doubleridge is a transgene-induced mutation characterized by polydactyly and syndactyly of the forelimbs. The transgene insertion maps to the proximal region of chromosome 19. During embryonic development of the mutant forelimb, delayed elevation and compaction of the apical ectodermal ridge (AER) produces a ridge that is abnormally broad and flat. Fgf8 expression persists in the ventral forelimb ectoderm of the mutant until E10.5. Strong expression of Fgf8 and other markers at the borders of the AER at E11.5 gives the appearance of a double ridge. At E11.5, apoptotic cells are distributed across the broadened ridge, but at E13.5, there is reduced apoptosis in the interdigital regions. The Shh expression domain is widely spaced at the posterior margin of the AER. The doubleridge AER is morphologically similar to that of En1 null mice, but the expression of En1 and Wnt7a is properly restricted in doubleridge, and the dorsal and ventral structures are correctly determined. doubleridge thus exhibits an unusual limb phenotype combining abnormal compaction of the AER with normal dorsal/ventral patterning. PMID- 12648497 TI - Contrasting activities of the aggregative and late PDSA promoters in Dictyostelium development. AB - Expression of the Dictyostelium PdsA gene from the aggregative (PdA) and late (PdL) promoter is essential for aggregation and slug morphogenesis, respectively. We studied the regulation of the PdA and PdL promoters in slugs using labile beta galactosidase (gal) reporter enzymes. PdL was active in prestalk cells as was also found with stable gal. PdA activity decreased strongly in slugs from all cells, except those at the rear. This is almost opposite to PdA activity traced with stable gal, where slugs showed sustained activity with highest levels at the front. PdA was down-regulated after aggregation irrespective of stimulation with any of the factors known to control gene expression. PdL activity was induced in cell suspension by cAMP and DIF acting in synergy. However, a DIF-less mutant showed normal PdL activity during development, suggesting that DIF does not control PdL in vivo. Dissection of the PdL promoter showed that all sequences essential for correct spatiotemporal control of promoter activity are downstream of the transcription start site in a region between -383 and -19 nucleotides relative to the start codon. Removal of nucleotides to position -364 eliminated responsiveness to DIF and cAMP, but normal PdL activity in prestalk cells in slugs was retained. Further 5' deletions abolished all promoter activity. This result also indicates that the induction by DIF and cAMP as seen in cell suspensions is not essential for PdL activity in normal development. PMID- 12648498 TI - XGef is a CPEB-interacting protein involved in Xenopus oocyte maturation. AB - XGef was isolated in a screen for proteins interacting with CPEB, a regulator of mRNA translation in early Xenopus development. XGef is a Rho-family guanine nucleotide exchange factor and activates Cdc42 in mammalian cells. Endogenous XGef (58 kDa) interacts with recombinant CPEB, and recombinant XGef interacts with endogenous CPEB in Xenopus oocytes. Injection of XGef antibodies into stage VI Xenopus oocytes blocks progesterone-induced oocyte maturation and prevents the polyadenylation and translation of c-mos mRNA; injection of XGef rescues these events. Overexpression of XGef in oocytes accelerates progesterone-induced oocyte maturation and the polyadenylation and translation of c-mos mRNA. Overexpression of a nucleotide exchange deficient version of XGef, which retains the ability to interact with CPEB, no longer accelerates oocyte maturation or Mos synthesis, suggesting that XGef exchange factor activity is required for the influence of overexpressed XGef on oocyte maturation. XGef overexpression continues to accelerate c-mos polyadenylation in the absence of Mos protein, but does not stimulate MAPK phosphorylation, MPF activation, or oocyte maturation, indicating that XGef may function through the Mos pathway to influence oocyte maturation. These results suggest that XGef may be an early acting component of the progesterone-induced oocyte maturation pathway. PMID- 12648499 TI - A study of regional gut endoderm potency by analysis of Cdx2 null mutant chimaeric mice. AB - Inactivation of Cdx2 by homologous recombination results in the development of forestomach epithelium at ectopic sites in pericaecal areas of the midgut of heterozygote mice. Local factors subsequently result in the secondary induction of tissues exhibiting an orderly sequence of tissue types between the ectopic forestomach tissue and the surrounding colon. Clonal analysis of this secondarily generated tissue using Y chromosome painting in chimaeric mice indicates that once differentiated to express Cdx2, host colonic epithelium can only form small intestinal-type epithelium, while Cdx2 mutant cells give rise to a succession of gastric-type tissue but never to a small intestine morphology. Our results indicate a difference in potency between forestomach and midgut precursor endodermal cells. PMID- 12648500 TI - Differentiation of nonbeating embryonic stem cells into beating cardiomyocytes is dependent on downregulation of PKC beta and zeta in concert with upregulation of PKC epsilon. AB - Cardiomyocyte differentiation overall has been analyzed in vivo and in vitro at the molecular level by homologous recombination, gene mutation studies, and by transgenics; however, the roles of many signal transduction mechanisms that drive this differentiation process are still not fully understood. One set of signal transduction components that has been studied in detail in mature, differentiated cardiomyocytes is the PKC isotype superfamily. However, while the function of each isotype is slowly being uncovered in adult cardiomyocytes, limited information persists concerning their function in the differentiation process of cardiomyocytes. To begin analyzing the function of specific PKC isotypes in the differentiation process, we employed an established model for differentiating ES cells into cardiomyocyte-positive embryoid bodies (EBs) in vitro. RT-PCR, Western analyses, and confocal microscopy all showed that the expression of specific PKC isotypes was significantly changed as ES cells differentiated into cardiomyocytes. More importantly, by using antagonists specific for each isotype we found that this change was a final step in the differentiation process. PKC beta and zeta downregulation served to promote differentiation (beating), while upregulation of PKC epsilon appeared to amplify differentiation (beating). Finally, melding classical tools (i.e., ionic exchange glass beads) with recently developed methods for differentiating ES cells creates a possible novel technique for investigating differentiation of ES cells into cardiomyocytes as well as other cell types. PMID- 12648501 TI - Lack of species-specificity in mammalian sperm chemotaxis. AB - Attraction of spermatozoa by way of chemotaxis to substances secreted from the egg or its surrounding cells has been demonstrated in marine species, amphibians, and mammals. This process is species- or family-specific in marine invertebrates: a chemoattractant for one marine species is usually not recognized by another species or by a member of another family. It is not known whether this selectivity is also the rule in other phyla. Furthermore, it is not at all obvious that such selectivity would be advantageous to species with internal fertilization. Here, using a directionality-based assay for chemotaxis, we studied in vitro the chemotactic response of human and rabbit spermatozoa to human, rabbit, and bovine egg-related factors. We found that spermatozoa from each of the two sources responded similarly well to egg-related factors obtained from any of the three species examined. These results indicate lack of chemotaxis related, species specificity between these species, suggesting that their sperm chemoattractants are common or very similar. The findings further suggest that mammals do not rely on species specificity of sperm chemotaxis for avoidance of interspecies fertilization. PMID- 12648502 TI - eIF4E/4E-BP dissociation and 4E-BP degradation in the first mitotic division of the sea urchin embryo. AB - The mRNA's cap-binding protein eukaryotic translation initiation factor (eIF)4E is a major target for the regulation of translation initiation. eIF4E activity is controlled by a family of translation inhibitors, the eIF4E-binding proteins (4E BPs). We have previously shown that a rapid dissociation of 4E-BP from eIF4E is related with the dramatic rise in protein synthesis that occurs following sea urchin fertilization. Here, we demonstrate that 4E-BP is destroyed shortly following fertilization and that 4E-BP degradation is sensitive to rapamycin, suggesting that proteolysis could be a novel means of regulating 4E-BP function. We also show that eIF4E/4E-BP dissociation following fertilization is sensitive to rapamycin. Furthermore, while rapamycin modestly affects global translation rates, the drug strongly inhibits cyclin B de novo synthesis and, consequently, precludes the completion of the first mitotic cleavage. These results demonstrate that, following sea urchin fertilization, cyclin B translation, and thus the onset of mitosis, are regulated by a rapamycin-sensitive pathway. These processes are effected at least in part through eIF4E/4E-BP complex dissociation and 4E-BP degradation. PMID- 12648503 TI - The role of Epstein-Barr virus and elevated levels of tumor necrosis factor in determining prognosis in Asian peripheral T-cell lymphomas. AB - Evidence of Epstein-Barr virus (EBV) infection in patients continues to be associated with the development of various lymphomas, and EBV genomic regions are detected in some lymphoma cells. Its role in causality and progression of disease versus the role of a passenger continues to be debated. Several reports of Asian T-cell lymphomas have also associated these diseases with evidence of EBV infections. Asian T-cell lymphomas are clinically distinct from Western T-cell lymphoma, and are also associated with increased levels of cytokine production, including tumor necrosis factor (TNF). The current study reports correlations of EBV + T-cell lymphoma with higher levels of TNF, and suggests some prognostic correlations. PMID- 12648504 TI - Commentary: effect of flavonoids on normal and leukemic cells. AB - The success of the phosphotyrosine kinase inhibitor imatinib mesylate (STI571, Gleevec has emphasized the significance of a growing understanding of tumor cell biology. The search has since been intensified to identify other candidate molecules in cancer cell-specific signaling transduction pathways whose disruption may result in similar therapeutic benefits. Flavonoids are potent inhibitors of cyclin-dependent kinases, but in addition also inhibit the activity of angiogenic mediators and induce apoptosis by mechanisms that are still not fully understand. In the current study by Liesveld et al., flavonoids are shown to have an antiproliferative and proapoptotic effect in leukemic cells. The implications of the results of this study on the activity of flavonoids in leukemias and their future development are being discussed. PMID- 12648505 TI - Empirical examination of the neutrophil criterion (>1500 microl(-1)) currently needed to declare CR in AML. AB - Currently, NCI guidelines require that the neutrophil count exceed 1500 before complete remission (CR) may be declared in acute myeloid leukemia (AML). We tested the empirical validity of this formulation by comparing event-free survival (EFS) in CR in (a). 305 patients who met the NCI criteria for CR and (b). 36 patients who met all the criteria for CR except that the neutrophil count at what we considered CR was 1000-1499 (lower neutrophil group) at which time they began post-remission therapy. EFS was statistically identical in both groups. We extended these findings to 752 patients, who met the NCI criteria for CR except that some may have had circulating blasts at "CR"; 82 of the 752 were in the lower neutrophil group. These data suggest that the minimum needed to declare CR in AML be changed from 1500 to 1000. PMID- 12648506 TI - The final phase in acute myeloid leukaemia (AML). A study on bleeding, infection and pain. AB - To increase the knowledge of the final phase of acute myeloid leukaemia (AML) a retrospective review of the medical and nursing records of 106 adult patients with AML who had died in 1995-1997 was made. A total of 27 patients were treated with curative intent at the time of death and 79 patients were prescribed palliative care. From the documentation, an evaluation of the frequency and severity of bleeding and pain episodes was made during their last week in life, and the occurrence of infection criteria was studied. Notations on bleeding were found in 44%, infection in 71% and pain in 76% of the patients. In 54% of the morphine administration days, no information on the effect of given morphine treatment was registered. To give AML patients in the final phase, the best possible treatment, skills in palliative medicine and palliative care are important. PMID- 12648507 TI - ABO incompatibility between donor and recipient and clinical outcomes in allogeneic stem cell transplantation. AB - We performed a retrospective, cohort study to evaluate the impact on recipient survival of ABO incompatibility between recipient and donor after allogeneic stem cell transplantation, primarily involving marrow-derived cells. No statistically significant difference was noted in survival for 153 patients with acute or chronic leukemia or myelodysplastic syndrome receiving ABO identical or ABO mismatched allografts. Five patients who had allografts that were bidirectionally incompatible (both donor cells and plasma incompatible) did have significantly poorer survival than the other recipients, similar to the experience reported in one other cohort study. However, these patients had other risks for mortality, including being older and receiving transplants from matched, unrelated donors. Our data do not support a significant role for ABO donor-recipient matching in allogeneic stem cell transplantation. PMID- 12648508 TI - High tumor necrosis factor-alpha levels in the patients with Epstein-Barr virus associated peripheral T-cell proliferative disease/lymphoma. AB - We have attempted to find out any relationships between circulating tumor necrosis factor (TNF)-alpha levels and Epstein-Barr virus (EBV) associated peripheral T-cell and NK-cell proliferative disease/lymphoma (PTPD/L) status. The distribution of TNF-alpha level was significantly higher (P<0.05) in patients than in controls. Patients carrying EBV genome in their peripheral T-cells showed higher TNF-alpha levels than the patients with EBV negative peripheral T-cells (P<0.001). Among patients whose peripheral T-cells were positive for EBV genome, TNF-alpha levels between the wild type LMP-1 gene carriers and the 30-bp deletion type LMP-1 gene carriers were compared and the wild type LMP-1 gene carrier group showed significantly higher TNF-alpha levels (P<0.01). As for the outcome of the patients and TNF-alpha levels, significant differences were observed between dead and alive with disease group (P<0.001), and dead and alive with complete remission group (P<0.01). Since circulating TNF-alpha levels in PTPD/L patients correlate with the disease and EBV infection status, it may be possible that monitoring of the TNF-alpha levels will be a useful prognostic marker. PMID- 12648509 TI - Bone marrow immunohistochemical studies of angiogenic cytokines and their receptors in myelofibrosis with myeloid metaplasia. AB - Bone marrow specimens from 11 patients with myelofibrosis with myeloid metaplasia (MMM) and seven normal controls were studied immunohistochemically to determine expression of transforming growth factor beta (TGF-beta), platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and corresponding receptors. Staining distribution and intensity for TGF-beta, PDGF, VEGF, TGF-beta type II receptor, a receptor for PDGF, and receptors for VEGF and bFGF were similar in patients and controls. Bone marrow from 10 MMM patients showed increased TGF-beta type I receptor (TGF-betaRI) expression in small vessel endothelial cells. Eight patient specimens had bFGF overexpression in megakaryocytes. Increased microvessel density and decreased concentration of bFGF-staining stromal cells accompanied these changes. Microvascular TGF-betaRI upregulation and bFGF overexpression by megakaryocytes may cause bone marrow microenvironmental changes in MMM patients. PMID- 12648510 TI - Monoclonal gammopathies in B-cell non-Hodgkin's lymphomas. AB - The association of monoclonal gammopathy (MG) with B-cell non-Hodgkin's lymphomas (NHL) is a well known phenomenon. The aim of the present work was to study the incidence, type of monoclonal component and prognostic significance of MG in a population of 255 cases with B-cell NHL. Among 255 evaluable patients with B-cell NHL, 145 were males and 110 females with a median age of 58 years (range 18-85). There were 166 patients with the various subtypes of aggressive (intermediate/high grade) NHL and 89 with the various subtypes of low risk. MG was detected in 44 patients (17.2%) with a median age of 61 years (range 23-79). There were 22 cases (8.6%) with IgG type (IgG/(k) 15, IgG/(lambda) 7), 4 cases (1.6%) with (IgA/(k) 3, IgA/(lambda) 1) and 18 cases (7.0%) with IgM (IgM/(k) 12 IgM/(lambda) 6). MG was found in 15.6% of the patients with aggressive NHL, while in low risk NHL the incidence was 20.2% (N.S.). The type of MG according to histological classification was as follows: Aggressive NHL: IgG 17 cases, IgA 2 cases, IgM 7 cases: low risk NHL: IgG 5 cases, IgA 2 cases, IgM 11 cases. The distribution of MG according to stage of the disease was as follows: stage I (4.5%), stage II (18%), stage III (6.8%) and stage IV (70.4%). The median survival of patients with aggressive NHL with MG was 17 months compared to 40 months of those without (P=0.22). Similarly the median survival of patients with low risk NHL and MG was 51.5 months compared to 38.5 months of those without (P=0.90). In conclusion MG was detected in 17.2% of cases with B-cell NHL. IgG-MG was more frequent in cases with aggressive NHL, while IgM in cases with low risk NHL. MG was mostly associated with advanced stage and had not any prognostic significance on survival. PMID- 12648511 TI - Involvement of DNA-dependent protein kinase in regulation of the mitochondrial heat shock proteins. AB - Since DNA-dependent protein kinase (DNA-PK) has been known to play a protective role against drug-induced apoptosis, the role of DNA-PK in the regulation of mitochondrial heat shock proteins by anticancer drugs was examined. The levels of basal and drug-induced mitochondrial heat shock proteins of drug-sensitive parental cells were higher than those of multidrug-resistant (MDR) cells. We also demonstrated that the development of MDR might be correlated with the increased expression of Ku-subunit of DNA-PK and concurrent down-regulation of mitochondrial heat shock proteins. The basal mtHsp70 and Hsp60 levels of Ku70(-/ ) cells, which were known to be sensitive to anticancer drugs, were higher than those of parental MEF cells, but conversely these mitochondrial heat shock proteins of R7080-6 cells over-expressing both Ku70 and Ku80 were lower than those of parental Rat-1 cells. Also, the mtHsp70 and Hsp60 levels of DNA-PKcs deficient SCID cells were higher than those of parental CB-17 cells. Our results suggest the possibility that mitochondrial heat shock protein may be one of determinants of drug sensitivity and could be regulated by DNA-PK activity. PMID- 12648512 TI - Flavonoid effects on normal and leukemic cells. AB - Quercetin and flavopiridol, both flavonoids which influence oxidative milieu, proliferation, and apoptosis of various cell types, were examined for their effects on acute myelogenous leukemic cells and normal progenitors. Both quercetin and flavopiridol inhibited the growth and viability of various acute myelogenous leukemia (AML) cell lines and AML blasts isolated afresh from patients with AML of various subtypes. The effects on inhibition of proliferation and decreased viability were also significant in normal CD34+ cells isolated from normal marrow donors. In certain AML cases, the effects of flavopiridol appeared to be mediated through activation of caspase 3, offering one possible mechanism for the apoptosis evident after exposure to flavopiridol as measured by annexin V expression. These flavonoid compounds might find use in various therapeutic settings in AML. PMID- 12648513 TI - A novel method for single cell detection of in situ telomerase or histone H3 in combination with clonal analysis by FISH. AB - A novel method for simultaneously detecting clonality by FISH, and presence of telomerase activity (telo+ cells) or histone H3 mRNA (H3+) in single cells from a mixed leukemic population is reported. The methods were validated using K562 cells mixed with peripheral blood granulocytes and bone marrow aspirate cells from newly diagnosed AML patients. Fifty patients with AML were analyzed for telo+ cells, while eight AML patients were analyzed for FISH-Telomerase and FISH H3+ during remission induction therapy. Our results demonstrate that: (1). changes in the leukemic populations during therapy could be followed; (2). a favorable response to chemotherapy occurred when there was a reduction in both the cytogenetically abnormal cells along with reduction in telo+ cells within this abnormal population; (3). reduction of either telo+ cells or FISH+ cells alone did not correlate with good response. H3+ could be detected in only 4% of the leukemic population, most of which were cytogenetically abnormal. These newly established methods allow sub-populations of cells to be followed during disease progression and treatment and to elucidate factors that give a specific clone proliferative advantage. PMID- 12648514 TI - Granulocyte-macrophage colony-stimulating factor: inhibitor of tumor necrosis factor-induced apoptosis. AB - Tumor necrosis factor (TNF) can induce proliferation as well as apoptosis in acute myeloid leukemia (AML)-derived cells. We have shown recently that these seemingly contradictory effects are based on the divergent capacities of the cells to produce granulocyte-macrophage colony-stimulating factor (GM-CSF) upon stimulation with TNF. Only those cells that produce GM-CSF survive the TNF attack and start growing. Here, we set out to elucidate the mechanisms of the antiapoptotic effect of GM-CSF. Protection from apoptosis was achieved by preincubating TF-1 cells with exogeneous GM-CSF. Cycloheximide prevented protection, indicating that GM-CSF might induce synthesis of antiapoptotic proteins. Regulation of protective genes was analyzed using cDNA expression arrays and the results were verified by Northern and Western blot analysis. This screen revealed the elevated expression of BCL-2, BCL-2A1, BAG-1 and TACE upon stimulation with GM-CSF. The major novelty of our study is that GM-CSF carries protective effects against TNF-induced apoptosis, not only against apoptosis induced by irradiation or cytokine-starvation. This protection requires de novo protein synthesis and is not-or at least not exclusively-the consequence of a direct crosstalk between the GM-CSF and TNF signaling pathways. PMID- 12648515 TI - Constitutively active mutant D816VKit induces megakayocyte and mast cell differentiation of early haemopoietic cells from murine foetal liver. AB - Mutations of Kit at position D816 have been implicated in mastocytosis, acute myeloid leukaemia and germ cell tumours. Expression of this mutant Kit in cell lines results in factor-independent growth, differentiation and increased survival in vitro and tumourigenicity in vivo. Mutant D816VKit and wild-type Kit were expressed in murine primary haemopoietic cells and grown in stem cell factor (SCF) or the absence of factors. Expression of D816VKit did not lead to transformation as assessed by a colony assay, but resulted in enhanced differentiation of cells when compared to control cells. D816VKit induced an increase in the number of cells differentiating along the megakaryocyte lineage in the absence of factors. SCF had an added effect with an increase in differentiation of mast cells. Expression of wild-type Kit in the presence of SCF also failed to cause transformation and induced differentiation of mast cells and megakaryocytes. We conclude that constitutive expression of D816VKit in primary haemopoietic cells is not a sufficient transforming stimulus but leads to the survival and maturation of cells whose phenotype is influenced by the presence of SCF. PMID- 12648516 TI - Serous retinal detachment in a case with chronic lymphocytic leukemia: no response to systemic and local treatment. AB - Leukemias are systemic hematopoietic neoplasias and not infrequently cause ocular findings. Serous retinal detachment (SRD) is one of these manifestations and even may be the first sign of the underlying leukemia. Here we reported a case with chronic lymphocytic leukemia (CLL) presenting with SRD and discussed the clinical importance and therapeutic options. PMID- 12648517 TI - Burkitt's leukemia with precursor B-cell immunophenotype and atypical morphology (atypical Burkitt's leukemia/lymphoma): case report and review of literature. AB - A 45-year-old male presented with fever, night sweats, splenomegaly, high LDH, high uric acid, and high blast count with atypical L3 morphology. Flow cytometry showed neoplastic clone consistent with pre B-cell immunophenotype, on the basis of lack of cytoplasmic immunoglobulins, surface immunoglobulins, and light chain immunoglobulins. Cytogenetic analysis revealed t(8;14)(q24; q32), the characteristic translocation of Burkitt's leukemia. A few similar case reports of Burkitt's leukemia with precursor B-cell immunophenotype have been described primarily in pediatric literature and usually with typical L3 morphology. This case recognizes this subset in adult ALL and emphasizes the need for comprehensive diagnostic analysis. PMID- 12648518 TI - Non-genomic effects of progestins--inhibition of cell growth and increased intracellular levels of cyclic nucleotides. AB - The anti-proliferative effect of progestins was studied in human transformed cell lines from the uterine cervix (C-4I, C33A and Me-180). Progestins caused a concentration-dependent inhibition of proliferation. The maximum tested concentration (2.6-3.2 microM) inhibited C-4I cell growth by the following order of potency: progesterone (56%) > medroxyprogesterone (38%) > megestrol acetate (25%). The sensitivity, expressed as I(25) (the concentration that caused 25% inhibition of growth), showed the same order: progesterone (7.7 nM) > medroxyprogesterone (78 nM) > megestrol acetate (570 nM). The intracellular levels of cGMP and cAMP were elevated and the cellular export of these cyclic nucleotides was inhibited by a similar order of potency. The C-4I cell line was devoid of progesterone-, estrogen-, androgen- and glucocorticoid-receptors. In addition, the antiprogestins mifepristone, onapristone and ZK-112993 did not block the anti-proliferative effect of progesterone. On the other hand, antiprogestins (2.3 nM) appeared to have some progesterone-like ("mimetic") activity with inhibition of C-4I cell growth; mifepristone (11%), onapristone (12%) and ZK-112993 (16%). The observed effects of progestins and antiprogestins on C-4I cells were also presented in C33A cells (16% androgen receptor positive) and Me-180 cells (22% progesterone receptor positive, 9% androgen receptor positive and 17% glucocorticoid receptor positive). This study suggests that a non-genomic mechanism contributes to the anti-proliferative effect of progestins. PMID- 12648519 TI - Evidence for two progesterone receptor binding sites in murine mammary carcinomas. AB - We have evaluated the progesterone receptor (PR) binding patterns in progestin dependent and -independent murine mammary carcinomas; all variants regress completely after antiprogestin treatment. These studies revealed the presence of a high affinity, low capacity-binding site (K(d): 43 +/- 9 pM; Q=9 +/- 3 fmol/mg protein) and of the classical lower affinity, high capacity-binding site (K(d): 9.2 +/- 4.2 nM; Q=376 +/- 64 fmol/mg protein). These sites could also be detected in uterus. Antiprogestins were able to bind to both sites. In vitro, medroxyprogesterone acetate (MPA) was stimulatory along a biphasic curve with two slopes, one at very low concentrations (EC(50): 1.5 +/- 0.7 fM) and the other at values compatible with the described K(d) for the PR (EC(50): 0.33 +/- 0.3 nM). PMID- 12648520 TI - The nuclear receptor corepressor (N-CoR) modulates basal and activated transcription of genes controlled by retinoic acid. AB - Variation in cell morphology and function is caused by differentiation. In myeloid differentiation, retinoid signaling, acting through heterodimers consisting of retinoic acid receptor and retinoid X receptor (RAR/RXR) plays a crucial part. The RAR/RXR heterodimers bind to naturally occurring response elements in the promoter regions of target genes, deciding whether the gene is to be transcribed or not. In the absence of the RAR-specific ligand all trans retinoic acid, RAR/RXR heterodimers are associated with the nuclear receptor corepressor N-CoR or the related SMRT. Here we show, using Western, far-Western and Northern blot techniques, that when the human monocytic cell line THP-1 is allowed to differentiate into macrophage-like cells the expression of N-CoR is down-regulated both at the protein and at the mRNA level. To investigate how this affects the transcriptional activity of retinoic acid response element (RARE) controlled genes, we performed transient transfection experiments in THP-1 and CV 1 cells. The results indicate that N-CoR functions not merely as a repressor of basal transcription, but rather as a modulator of both basal and ligand-activated transcription of genes controlled by RAR/RXR heterodimers in a dose-dependent manner. PMID- 12648522 TI - Limited species differences in estrogen receptor alpha-medicated reporter gene transactivation by xenoestrogens. AB - Estrogen receptors (ERs) play an important role in estrogen function. However, it is well known that there are species differences in amino acid sequences of the ligand binding domains. Here, we report on the analysis of species differences in ER-dependent transactivation with some chemicals using reporter gene assays. Full length ER cDNAs from human, rat, chicken, alligator (Caiman), whiptail lizard, African clawed frog and rainbow trout were prepared from hepatic mRNA by the RT PCR method and inserted into expression plasmids. Both expression and reporter plasmids were transiently transfected into HeLa cells, and then the estrogenic effects of chemicals were analyzed in terms of induction of luciferase activity. No species differences in transactivation were found among human, rat, chicken, alligator, whiptail lizard and African clawed frog ERs. However, thermo-dependent alteration in susceptibility to 17-beta-estradiol was observed with the rainbow trout ER because of thermo-dependence of estrogen binding. PMID- 12648521 TI - 17 beta-estradiol- and 4-hydroxytamoxifen-induced transactivation in breast, endometrial and liver cancer cells is dependent on ER-subtype, cell and promoter context. AB - The pattern of transcriptional activation by 17beta-estradiol (E2) and 4 hydroxytamoxifen (4-OHT) was determined in ZR-75 and MDA-MB-231 breast, ECC1 and HEC1A endometrial and HepG2 liver cancer cell lines cotransfected with E2 responsive constructs and wild-type estrogen receptor alpha (ER alpha) or ER beta (ER beta) or variant forms of ER alpha expressing activation function 1, AF1 (ER alpha-AF1) or activation function 2, AF2 (ER alpha-AF2). The E2-responsive constructs contained promoter inserts from the human complement C3 (pC3), human cathepsin D (pCD) and rat creatine kinase B (pCKB) genes. Minimal ER beta dependent transactivation (<2.5-fold induction) was observed for E2 only in ECC1 and MDA-MB-231 cells transfected with pCKB or pC3, whereas 4-OHT was inactive as an ER beta agonist for all promoters in the four cell lines. The ER alpha agonist and/or antagonist activities for E2 and 4-OHT were highly variable and the transactivation was dependent on ER subtype, ER alpha variant expressed, gene promoter, and cell context. For example, E2 did not activate pCD in HepG2 cells transfected with wild-type or variant ER alpha, whereas E2 activated reporter gene activity in the four endometrial and breast cancer cell lines transfected with ER alpha and pCD, pCKB or pC3. Hormone activation of these constructs by ER alpha-AF1 or ER alpha-AF2 was highly variable among the different cell lines and even in the same cell line transfected with the three E2-responsive constructs. Similar variability was observed for 4-OHT. For example, 4-OHT activates pC3 in HepG2 cells transfected with ER alpha or ER alpha-AF1, and pCKB in HEC1A cells. However, AF1-dependent activation by 4-OHT is not observed for pCKB in ECC1 cells or for pC3 and pCD in HEC1A or ECC1 endometrial cancer cells. The results of this study suggest that transcriptional activation by E2 and 4-OHT induces recruitment of different transcription factor complexes that are dependent on the cell type and also the gene promoter. PMID- 12648523 TI - Molecular communication between androgen receptor and general transcription machinery. AB - The androgen-androgen receptor (AR) signaling pathway plays a key role in proper development and function of male reproductive organs. Like other transcriptional regulators, AR may communicate with the general transcription machinery on the core promoter to exert its function as a transcriptional modulator. The molecular communication between AR and the general transcription machinery may be achieved either by the direct protein-protein interaction between AR and the general transcription machinery or by the indirect interaction mediated by coregulators. Analyses of AR-mediated transcription suggest that the orchestrated interaction of AR with the transcription factors IIF (TFIIF) and IIH (TFIIH), and positive transcription elongation factor b (P-TEFb), may increase efficiency of transcriptional elongation from the androgen target genes, such as prostate specific antigen (PSA). Based on studies so far, AR may regulate transcription not by enhanced assembly of preinitiation transcription complex but by regulating promoter clearance and elongation stage of transcription. PMID- 12648524 TI - Effect of 2-methoxyestradiol on the growth of methyl-nitroso-urea (MNU)-induced rat mammary carcinoma. AB - The present study investigated the influence of the endogenous estradiol metabolite 2-methoxyestradiol (2ME) on the growth of methyl-nitroso-urea (MNU) induced mammary carcinoma in the rat. 2ME was administered by means of subcutaneously implanted osmotic pumps for a period of 4 weeks. The dosages of 2ME were 1 and 5mg/kg per day, the control animals received saline. At the low dosage of 2ME a stimulation of tumor growth was observed, whereas at the high dosage an inhibition was found. The urinary excretion of 15 estradiol metabolites revealed that 2ME triggered strong changes in estrogen metabolism in the organism. Our data show that 2ME may elicit both stimulation and inhibition of tumor growth depending on the dosage used, a fact which should be considered in case of therapeutic use. PMID- 12648525 TI - Molecular targets of 1,25(OH)2D3 in HC11 normal mouse mammary cell line. AB - Our aim was to determine the molecular targets involved in the antiproliferative effects of 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)), in a normal murine mammary epithelial cell line, HC11. Among the early response genes analyzed, c myc, junB, junD, c-jun, c-fos, fosB, fra, as well as max, mad1-4, sin3, only c jun and fra-2 mRNAs were up-regulated after 1,25(OH)(2)D(3) exposure. Cyclin C was reduced and cyclin A2 and E were slightly enhanced; however, cyclins D1, D3, B1, B2, F, G1, G2, I and H, as well as TGF beta 1, TGF beta 3, T beta RI and T beta RII transcripts were not modulated by 1,25(OH)(2)D(3). Although p27(KIP1) protein content was unchanged, enhancement of p21(WAF1/CIP1) low basal levels in cell extracts and IGFBP-3 abundance on the culture medium was detected after 1,25(OH)(2)D(3) induction. Using differential display analysis, we identified eight down-modulated clones in exposed cells: 26S proteasome non-ATPase subunit Pad1, ubiquitin-conjugating enzyme Ube2i, extracellular proteinase inhibitor Expi or Wdnm1, cytochrome-c oxidase Cox7c, microtubule-associated protein-1 light chain-3 (Map1lc3), nascent-associated complex alpha Naca, transforming acidic coiled-coil Tacc3, stearoyl-CoA desaturase (Scd), keratin 6 alpha, and 1 up regulated, fork head transcription factor Hfh-1L. Hence, the antiproliferative effect of 1,25(OH)(2)D(3) seems associated to enhancement of c-jun, Fra-2, IGFBP3 and p21(WAF1/CIP1). Decreased Pad1 and Ube2i might account for increased stability of cell cycle inhibitory proteins while reduced Wdnm1, Tacc3 and Scd might be secondary to accumulation of cells in G0/G1 phase. PMID- 12648526 TI - Binding of ovarian steroids to erythrocytes in patients with sickle cell disease; effects on cell sickling and osmotic fragility. AB - Ovarian steroids appear to influence the manifestations of sickle cell disease (SCD); oestrogens can adversely affect erythrocyte function, whereas progestogens may inhibit sickling and decrease the osmotic fragility of erythrocytes. The aims of the present studies were: (i) to characterise the binding of oestradiol and progesterone to erythrocytes from women with HbSS, HbSC and HbAA genotypes; (ii) to investigate whether steroids modulate susceptibility to sickling or osmotic fragility of HbSS and HbAA erythrocytes. Erythrocytes were incubated for 1h with [3H]-steroids at 4 and 37 degrees C. Binding of both oestradiol and progesterone was independent of temperature and steroid concentration, but was decreased by sequential "washing" of erythrocytes in fresh incubation buffer. Binding capacity was 80 +/- 6% greater for oestradiol (versus progesterone) in all three genotypes, and binding of both steroids was decreased by > or = 70% in HbSS erythrocytes compared to HbSC or HbAA erythrocytes. Pre-incubation of erythrocytes with 35 microM oestradiol or 30 microM progesterone had no significant effect on susceptibility of HbSS and HbAA erythrocytes to sickling, or on osmotic fragility. We conclude that both oestradiol and progesterone bind in a low affinity, non-saturable manner to erythrocytes with decreased binding in cells from women with HbSS. However, steroid binding does not affect susceptibility to sickling or osmotic fragility irrespective of haemoglobin genotype. PMID- 12648527 TI - Influence of stage of the reproductive cycle and estradiol on thymus cell antigen presentation. AB - The objective of this study was to determine whether thymus cells present antigen and if endocrine balance influences antigen presentation. We report here that antigen presenting cells (APC) from the thymus glands of male and female rats, when incubated with ovalbumin (OVA)-specific T cells and OVA, are functionally able to present antigen via MHC class II. To determine whether antigen presentation in the thymus is under hormonal control, tissues from female rats at different stages of the estrous cycle were analyzed. Antigen presentation was higher at estrus and proestrus than that seen at diestrus when estradiol levels are low. Estradiol given to ovariectomized animals for 3 days stimulated antigen presentation by adherent thymus cells compared to saline controls. Flow cytometry studies indicated that the adherent thymus cell preparations consisted of DC, T cells, B cells and cells of the myeloid lineage all of which expressed MHC class II, as did a small population of non-leukocytes. Antibody neutralization studies indicated that thymus cell antigen presentation involves the expression of transmembrane proteins B7.1 and B7.2. These studies demonstrate that sex hormones play a central role in regulating antigen presentation in the thymus. PMID- 12648528 TI - Differential expression of P450 aromatase during gonadal sex differentiation and sex reversal of the newt Pleurodeles waltl. AB - A better understanding of vertebrate sexual differentiation could be provided by a study of models in which genetic sex determination (GSD) of gonads can be reversed by temperature. In the newt Pleurodeles waltl, a P450 aromatase cDNA was isolated from adult gonads, and the nucleotide or deduced amino acid sequences showed a high level of identity with various vertebrate species. In adults, aromatase expression was found in gonads and brain. In developing gonads, the expression was found to fit with the thermo-sensitive period (TSP) and was detected in both ZZ and ZW larvae, as well as in ZW submitted during the whole TSP to a masculinizing temperature. In the latter individuals, in situ hybridization and semi quantitative RT-PCR showed that, at the end of TSP, aromatase expression was at the same level than in normal ZZ larvae and was significantly lower than in normal ZW ones. Furthermore, temperature-induced down regulation did not occur when heating was performed at the end of TSP. Our results confirm the importance of aromatase regulation in female versus male differentiation and demonstrate that a down regulation of aromatase expression is involved in the process of sex reversal. PMID- 12648529 TI - Calreticulin inhibits the MEK1,2-ERK1,2 pathway in alpha 1-adrenergic receptor/Gh stimulated hypertrophy of neonatal rat cardiomyocytes. AB - In cardiac myocytes, stimulation of alpha(1)-adrenoceptor (AR) leads to a hypertrophic phenotype. The G(h) protein (transglutaminase II, TGII) is tissue type transglutaminase and transmits the alpha(1B)-adrenoceptor signal with GTPase activity. Recently, it has been shown that the calreticulin (CRT) down-regulates both GTP binding and transglutaminase activities of TGII. To elucidate whether G(h) mediates norepinephrine-stimulated intracellular signal transductions leading to activation of extracellular signal-regulated kinases (ERKs) and neonatal rat cardiomyocyte hypertrophy, we examined the effects of G(h) on the activation of ERKs and inhibitory effects of CRT on alpha(1)-adrenoceptor/G(h) signaling. In neonatal rat cardiomyocytes, norepinephrine-induced ERKs activation was inhibited by an alpha(1)-adrenoceptor blocker (prazosin), but not by an beta adrenoceptor blocker (propranolol). Overexpression of the G(h) protein stimulated norepinephrine-induced ERKs activation, which was inhibited by alpha-adrenoceptor blocker (prazosin). Co-overexpression of G(h) and CRT abolished norepinephrine induced ERKs activation. Taken together, norepinephrine induces hypertrophy in neonatal rat cardiomyocytes through alpha(1)-AR stimulation and G(h) is partly involved in norepinephrine-induced MEK1,2/ERKs activation. Activation of G(h) mediated MEK1,2/ERKs was completely inhibited by CRT. PMID- 12648530 TI - Expression of perivitelline membrane glycoprotein ZP1 in the liver of Japanese quail (Coturnix japonica) after in vivo treatment with diethylstilbestrol. AB - Avian perivitelline membrane, an oocyte extracellular matrix homologous to the zona pellucida in mammals or chorion in fish, is composed of at least two glycoproteins. Previous studies have indicated that one of the components, a glycoprotein homologous to mammalian ZPC, is produced in the granulosa cells of the developing follicles of quail ovary on stimulation with testosterone. However, little is known about the molecular biology of the other component of the avian perivitelline membrane, ZP1, and information about gene expression is particularly lacking. We have cloned the ZP1 in Japanese quail and examined its gene expression. A cDNA encoding quail ZP1 was isolated from the livers of mature females using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. It encoded a 934-amino acid protein that showed greatest homology (87.8% identity) with the chicken ZP1. RT-PCR amplification indicated that the ZP1 mRNA in the liver was restricted to mature laying females. The expression of ZP1 mRNA was stimulated by in vivo treatment with diethylstilbestrol in immature females as well as males. These results suggested that androgens and estrogens coordinately regulate the formation of quail perivitelline membrane proteins. In addition, the use of ZP1 transcriptional induction in males or immature females as a biological marker of environmental estrogens is discussed. PMID- 12648531 TI - Tetranortriterpenoid derivatives from Turraea parvifolia (Meliaceae). AB - The methanol extract of the seeds of Turraea parvifolia has yielded seven novel triterpenoid derivatives: 12alpha-acetoxyazadironolide, turraparvin A (12alpha acetoxy-7alpha,23-dihydroxy-24,25,26,27-tetranor-3-oxoapotirucalla-1,14,20(22) trien-21,23-olide), turraparvin B (12alpha-acetoxy-7alpha,21-dihydroxy 24,25,26,27-tetranor-3-oxoapotirucalla-1,14,20(22)-trien-23,21-olide), turraparvin C (7alpha,12alpha-diacetoxy-21-hydroxy-24,25,26,27-tetranor-3 oxoapotirucalla-1,14,20(22)-trien-23,21-olide), 11-epi-21-hydroxytoonacilide, 11 epi-23-hydroxytoonacilide and turraparvin D (12alpha-acetoxy-7alpha-hydroxy 24,25,26,27-tetranor-3-oxoapotirucalla-1,14,20(22)-trien-21,23-lactam). PMID- 12648533 TI - Sesquiterpene lactones from glandular trichomes of Viguiera radula (Heliantheae; Asteraceae). AB - In addition to known compounds, the floral parts of Viguiera radula afforded two new sesquiterpene lactones. All compounds were detected in glandular trichomes, which were micromechanically collected from the anther appendages and analyzed by HPLC. Structure identification was performed by NMR and MS techniques. PMID- 12648534 TI - Terpenes from Inula verbascifolia. AB - The aerial parts of Inula verbascifolia afforded two new xanthanes and a new germacranolide derivative, together with the known compounds inusoniolide, 4-O dihydroinusoniolide and 9beta-hydroxyparthenolide. The structures were determined by spectral methods (IR, HRMS,1H NMR, 13C NMR, DEPT, 1H-1H COSY, HMQC and HMBC). PMID- 12648532 TI - Ecdysteroids and other constituents from Sida spinosa L. AB - Two compounds (3 and 10) were isolated from the aerial parts of Sida spinosa L. Their structures have been established as glyceryl-1-eicosanoate and 20-hydroxy, 24-hydroxymethylecdysone by 1D and 2D-NMR techniques. In addition 12 known compounds (1, 2, 4-9 and 11-14) have been isolated and identified. PMID- 12648535 TI - Eudesmane derivatives and other constituents from Saussurea parviflora. AB - A survey of the whole plant of Saussurea parviflora afforded three compounds 11,12,13-trihydroxy-4(15),8-eudesmadiene-9-one, eudesman-8beta,12-olide-1-O-beta D-glucoside and 1beta,3beta-dihydroxyursa-9(11),12-diene-3-octadecanoate, as well as 13 known compounds. Their structures were elucidated on the basis of spectral evidence, especially by using NMR spectroscopic techniques. In addition, encelin exhibited effective antitumor activity on L02, SMMC-7721 and HO-8910 cells. PMID- 12648536 TI - Further saponins from Taverniera aegyptiaca. AB - From the saponin fraction of the total methanolic extract of the dried root and stem barks of Taverniera aegyptiaca Boiss, six new triterpenoidal saponins of oleanane type were isolated and identified as 28-methyl serratagenate-3-beta-O beta-xylopyranosyl (1-->2)-beta-glucopyranoside (2), 28-methyl serratagenate 3 beta-O-alpha-rhamnopyranosyl (1-->2)-beta-glucopyranoside (3), 3beta-O-alpha rhamnopyranosyl (1-->2) beta-glucopyranosyl-olean-11,13(18)-dien-1beta, 3beta, 22beta-triol (4), 3beta-O-beta-glucopyranosyl (1-->2)-beta glucuronopyranosylolean-11,13(18)-dien-1beta,3beta,22beta-triol (5), 3beta-O-beta xylopyranosyl(1-->2)-beta-glucuronopyranosylolean-11,13(18)-dien 1beta,3beta,22beta-triol (6), 3beta-O-alpha-rhamnopyranosyl (1-->2)-beta glucuronopyranosylolean-11,13(18)-dien-1beta, 3beta, 22beta-triol (7) together with the known oleanolic acid 3-beta-O-beta-glucoside (1). The identification of the isolated compounds was done on the basis of chemical and spectral evidences. PMID- 12648537 TI - Ceriopsins F and G, diterpenoids from Ceriops decandra. AB - Chemical examination of the ethyl acetate solubles of the CH(3)OH:CH(2)Cl(2) (1:1) extract of the roots of Ceriops decandra collected from Kauvery estuary resulted in the isolation of two more diterpenoids, ceriopsins F and G (1-2) and five known compounds, ent-13-hydroxy-16-kauren-19-oic acid (steviol, 3), methyl ent-16beta,17-dihydroxy-9(11)-kauren-19-oate (4), ent-16beta,17-dihydroxy-9(11) kauren-19-oic acid (5), ent-16-oxobeyeran-19-oic acid (isosteviol, 6), 8,15R epoxypimaran-16-ol (7). The structures of the new diterpenoids were elucidated by a study of their physical and spectral data as methyl ent-13,17-epoxy-16 hydroxykauran-19-oate (1) and ent-16-oxobeyeran-19-al (2). PMID- 12648538 TI - Guaiane sesquiterpenes from Amoora rohituka. AB - The petroleum ether extract of the stem bark of Amoora rohituka afforded two novel guaiane-derived sesquiterpenoids, 6beta,7beta-epoxyguai-4-en-3-one (1) and 6beta,7beta-epoxy-4beta,5-dihydroxyguaiane (2). The structures of 1 and 2 were determined by extensive NMR and MS analyses and by comparison of their spectral data with related compounds. The relative stereochemistry of the asymmetric centers in 1 and 2, except at C-5 of 2, were determined by selective 1D-NOESY experiments. PMID- 12648539 TI - Anthocyanins from red onion, Allium cepa, with novel aglycone. AB - Four anthocyanins with the same novel 4-substituted aglycone, carboxypyranocyanidin, have been isolated from acidified, methanolic extracts of the edible scales as well as from the dry outer scales of red onion, Allium cepa L. The structures of 1 and 2 were identified as the 3-O-beta-glucopyranoside and 3-O-(6"-O-malonyl-beta-glucopyranoside) of 5-carboxypyranocyanidin, respectively. This aglycone, 5-carboxy-2-(3,4-dihydroxyphenyl)-3,8-dihydroxy-pyrano[4,3,2-de]-1 benzopyrylium, is with exception of the substitution pattern on the phenyl ring similar to carboxypyranomalvidin (vitisidin A) recently isolated from red wines. In addition to 1 and 2, two analogues of 2 methylated at the terminal carboxyl group of the acyl moiety (3) or at the aglycone carboxyl (4), respectively, were also identified. These latter compounds are most probably formed by esterification of 2 with the solvent (acidified methanol) during the isolation process. The structures were elucidated by 2D NMR spectroscopy and LC-MS. PMID- 12648540 TI - Antibacterial bromophenols from the marine red alga Rhodomela confervoides. AB - Two bromophenols, together with three known compounds, were isolated from the methanolic extract of the marine alga, Rhodomela confervoides. By means of MS and NMR spectroscopic analyses, they were identified as 3-bromo-4-[2,3-dibromo-4,5 dihydroxyphenyl] methyl-5-(hydroxymethyl) 1,2-benzenediol (1) and 3-bromo-4-[2,3 dibromo-4,5-dihydroxyphenyl] methyl-5- (ethoxymethyl) 1,2-benzenediol (2). Three known compounds were also isolated, namely 3-bromo-4-[2,3-dibromo-4,5 dihydroxyphenyl] methyl-5-(methoxymethyl) 1,2-benzenediol (3), 4,4'- methylenebis [5,6-dibromo-1,2-benzenediol] (4) and bis (2,3-dibromo-4,5-dihydroxybenzyl) ether (5). Compound 5 was the most active against five strains of bacteria with the MIC less than 70 microg/ml, while compounds 2, 3 and 4 exhibited moderate activity. PMID- 12648541 TI - Chalcones from the seed of Cedrelopsis grevei (Ptaeroxylaceae). AB - The seed of Cedrelopsis grevei (Ptaeroxylaceae) has yielded the known compounds uvangoletin, 5,7-dimethylpinocembrin, cardamonin, flavokawin B, 2' methoxyhelikrausichalcone, and the novel prenylated chalcones, cedreprenone and cedrediprenone. Cedridiprenone has been shown to exhibit superoxide scavenging properties. PMID- 12648542 TI - Acetylated phenolic glycosides from Harpagophytum procumbens. AB - Two acetyl phenolic glycosides, 6-acetylacteoside and 2,6-diacetylacteoside, were obtained from commercially available secondary roots of Harpagophytum procumbens and were identified using spectroscopic methods. PMID- 12648543 TI - Antiviral flavonoids from the root bark of Morus alba L. AB - A prenylated flavonoid, moralbanone, along with seven known compounds kuwanon S, mulberroside C, cyclomorusin, eudraflavone B hydroperoxide, oxydihydromorusin, leachianone G and alpha-acetyl-amyrin were isolated from the root bark of Morus alba L. Leachianone G showed potent antiviral activity (IC(50) = 1.6 microg/ml), whereas mulberroside C showed weak activity (IC(50) = 75.4 microg/ml) against herpes simplex type 1 virus (HSV-1). Their structures were elucidated by spectroscopic methods. PMID- 12648544 TI - 6-Hydroxypelargonidin glycosides in the orange-red flowers of Alstroemeria. AB - Two 6-hydroxypelargonidin glycosides were isolated from the orange-red flowers of Alstroemeria cultivars, and determined to be 6-hydroxypelargonidin 3-O-(beta-D glucopyranoside) and 3-O-[6-O-(alpha-L-rhamnopyranosyl)-beta-D-glucopyranoside], respectively, by chemical and spectroscopic methods. In addition, five known anthocyanidin glycosides, 6-hydroxycyanidin 3-malonylglucoside, 6-hydroxycyanidin 3-rutinoside, cyanidin 3-malonylglucoside, cyanidin 3-rutinoside and pelargonidin 3-rutinoside were identified in the flowers. PMID- 12648545 TI - Eryvarins F and G, two 3-phenoxychromones from the roots of Erythrina variegata. AB - Two 3-phenoxychromones, eryvarins F and G, were isolated from the roots of Erythrina variegata. Their structures were established to be 3-(2,4 dihydroxyphenoxy)-7-hydroxy-6,8-di(3,3-dimethylallyl)chromen-4-one and 3-(2,4 dihydroxyphenoxy)-8-(3,3-dimethylallyl)-2,2-dimethylpyrano[5,6:6,7]chromen-4-one on the basis of spectroscopic and chemical evidence. Eryvarins F and G are unusual 3-phenoxychromone derivatives with two isoprenoid groups. PMID- 12648546 TI - Acylated phenolic glycosides from Solenostemma argel. AB - From the aerial parts of Solenostemma argel, four new acylated phenolic glycosides sinapyl alcohol 9-O-feruloyl-4-O-alpha-rhamnopyranosyl-(1-->2)-beta glucopyranoside, solargin I (1), sinapyl alcohol 9-O-caffeoyl-4-O-alpha rhamnopyranosyl-(1-->2)-beta-glucopyranoside, solargin II (2), sinapyl alcohol 9 O-feruloyl-4-O-alpha-rhamnopyranosyl-(1-->2)-alpha-rhamnopyranosyl-(1-->2)-beta glucopyranoside, solargin III (3) and sinapyl alcohol 9-O-caffeoyl-4-O-alpha rhamnopyranosyl-(1-->2)-alpha-rhamnopyranosyl-(1-->2)-beta-glucopyranoside, solargin IV (4) have been isolated. The structures of the isolated compounds were verified by means of MS and NMR spectral analyses. PMID- 12648547 TI - Muscanone: a 3-O-(1", 8", 14"-trimethylhexadecanyl)naringenin from Commiphora wightii. AB - A new antifungal flavanone, muscanone (1), was isolated along with known naringenin (2) from Commiphora wightii (Arn.) Bhandari (Burseraceae) by directing the fractionation of an EtOH extract of the air-dried trunk of C. wightii with microbial sensitivity assay. The structures of 1 and 2 were determined from EIMS, HREIMS, DEPT, 1H-1H COSY, HSQC and HMBC spectral data. Muscanone (1) was identified as 3-O-(1", 8",14"-trimethylhexadecanyl)naringenin and was found to be active against Candida albicans. The isolation, structure elucidation, NMR spectral assignments, and bioactivities of 1 and 2 are reported. PMID- 12648548 TI - Ipobscurines C and D: macrolactam-type indole alkaloids from the seeds of Ipomoea obscura. AB - Separation of the methanolic seed extract of Ipomoea obscura afforded five indole alkaloids, three of them (ipobscurines B-D) being new natural products of a unique structural type characterized as serotonin hydroxycinnamic acid amide-type conjugates with a second phenylpropanoid moiety forming an ether with the 5-OH position of the indole nucleus. Due to an oxidative phenolic coupling between the two phenylpropanoid moieties of the supposed precursor ipobscurine B two 21 membered macrolactams with a phenol ether partial structure are formed: the trans cis isomers ipobscurines C and D. Their structures were established on the basis of spectral data. Moreover, total synthesis of the racemic erythro- and threo ipobscurine B 4',4"-dimethyl ethers and the comparison with the corresponding derivative of natural (-)-ipobscurine B proved an erythro configuration of the latter. PMID- 12648549 TI - Tetrahydroisoquinoline alkaloids and 2-deoxyribonolactones from Aristolochia arcuata. AB - 2-Deoxyribonolactones and four tetrahydroisoquinoline alkaloids were isolated from the acetone extract of the leaves of Aristolochia arcuata Mast., together with pinitol, sequoyitol, glycerol, fructose, sucrose, eupomatenoid-7, salsolinol, and 6,7-dihydroxy-1,1-dimethyl-1,2,3,4-tetrahydroisoquinoline. Their structures were determined on the basis of spectroscopic methods, mainly using 1H, 13C, 15N, and 31P NMR. PMID- 12648550 TI - A flavone and an unusual 23-carbon terpenoid from Andrographis paniculata. AB - Phytochemical investigation of the roots and aerial parts of Andrographis paniculata Nees yielded a new flavone, 5-hydroxy-7,2',6'-trimethoxyflavone and an unusual 23-carbon terpenoid, 14-deoxy-15-isopropylidene-11,12 didehydroandrographolide together with five known flavonoids and four known diterpenoids. The structures of these compounds were determined on the basis of spectral and chemical studies. PMID- 12648551 TI - Further constituents from Caralluma negevensis. AB - Two new megastigmane glycosides (1 and 2) and two new flavone glycosides (3 and 4) were isolated from the methanol extract of the whole plant of Caralluma negevensis Zohary (Asclepiadaceae). The structures of the isolated compounds were characterized as (9R)-2beta,9-dihydroxymegastigma-4,7-dien-3-one-9-O-alpha-L rhamnopyranosyl-(1-->6)-beta-D-glucopyranoside (1), 2beta,9-dihydroxymegastigma-4 en-3-one 9-O-alpha-L-rhamnopyranosyl-(1-->6)-beta-D-glucopyranoside (2), luteolin 3'-O-beta-D-glucopyranoside-4'-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D glucopyranoside (3), and luteolin 3',4'-di-O-beta-D-glucopyranoside (4). The structures of the isolated compounds were established on the basis of spectral evidence and chemical transformation. PMID- 12648552 TI - Biochemical characterization of blood orange, sweet orange, lemon, bergamot and bitter orange. AB - This paper reports on the composition of aroma compounds and fatty acids and some physico-chemical parameters (juice percentage, acidity and total sugars) in five varieties of citrus: blood orange, sweet orange, lemon, bergamot and bitter orange. Volatile compounds and methyl esters have been analyzed by gas chromatography. Limonene is the most abundant compound of monoterpene hydrocarbons for all of the examined juices. Eighteen fatty acids have been identified in the studied citrus juices, their quantification points out that unsaturated acids predominate over the saturated ones. Mean concentration of fatty acids varies from 311.8 mg/l in blood orange juice to 678 mg/l in bitter orange juice. PMID- 12648554 TI - X-linked amelogenesis imperfecta may result from decreased formation of tyrosine rich amelogenin peptide (TRAP). AB - Amelogenesis imperfecta (AI) is a group of inherited disorders with defective tooth enamel formation caused by various gene mutations. One of the mutations substitutes a cytidine for an adenine in exon 6 of the X-chromosomal amelogenin gene, which results in a proline to threonine change in the expressed amelogenin. This transformation is four amino acids N-terminal to the cleavage site for enamel matrix metalloproteinase-20 (MMP-20) in amelogenin. MMP-20 releases the tyrosine rich amelogenin peptide (TRAP) from amelogenin. This study evaluated the rate at which MMP-20 hydrolyses mutated amelogenin relative to unmutated amelogenin. A full-length recombinant human amelogenin and a mutated amelogenin with a substitution of proline by threonine were expressed and purified by ammonium sulphate precipitation and reverse phase HPLC. Recombinant metalloproteinase-20 (rMMP-20) was used to digest the recombinant proteins, which resulted in fragments with a mass predicted for TRAP. The proteolytic site was also modelled as substrates by two synthetic peptides, SYGYEPMGGWLHHQ and SYGYETMGGWLHHQ, selected from residues 36 to 49 of the amino acid sequence for amelogenin and the respective X-linked amelogenin mutant. These two peptides were labelled at their N- and C-termini respectively by using rhodamine and biotin. After digestion with MMP-20, the truncated peptides were separated by avidin labelled magnetic Dynal beads and were identified by mass spectrometry. These results demonstrated that both oligopeptides were cleaved between tryptophan and leucine, matching the TRAP cutting site found in tooth enamel. Enzyme kinetics showed that the k(cat)/K(m) of rMMP-20 against the unmutated amelogenin peptide was 21 times greater than that against the mutated peptide. This study suggests that the reduced rate of TRAP formation by a single amino acid substitution alters enamel matrix hydrolysis by MMP-20, which may result in amelogenesis imperfecta. PMID- 12648555 TI - Expression of vascular endothelial growth factor in human dysfunctional temporomandibular joint discs. AB - A high density of blood vessels is found in specimens of temporomandibular joint (TMJ) disc at any stage of internal derangement of the joint, but the factors responsible for angiogenesis in the disc have not been described. The purpose here was to investigate, in human TMJ discs, the expression of vascular endothelial growth factor (VEGF), a multifunctional cytokine that contributes to angiogenesis. Specimens, free of significant morphological alterations and with varying degrees of disc tissue degeneration/regeneration, were studied by immunohistochemistry for VEGF in order to correlate immunohistochemical with histopathological findings. In normal discs and discs with minor pathological changes, fibroblast-like cells, fibrochondrocytes and chondrocyte-like cells were either not or only weakly immunostained by VEGF antibody. In disc specimens from internal derangement of the TMJ with significant tissue degeneration/regeneration, VEGF was consistently expressed. In these specimens, immunoreaction products for VEGF were observed both in the disc and in the endothelial cells of newly formed vessels. This VEGF immunolocalization is consistent with the stimulation of angiogenesis and the morphogenesis and differentiation of chondrocytes. Therefore VEGF expression by disc chondrocyte like cells might reflect the action of the cytokine as an inducer of angiogenesis and as an autocrine signal for cells of the chondrogenic lineage. PMID- 12648556 TI - Chewing behaviour and bolus formation during mastication of meat with different textures. AB - During chewing, meat is mashed under compression and shear bite forces whilst saliva is incorporated. The resulting mixture is shaped into a cohesive bolus by agglomeration of small particles, and triggers a swallow. This study aimed to investigate the relationship between chewing behaviour and bolus formation of meat with different textures. Twenty-five consenting young adults participated in this study. Electromyographic activity was recorded from surface electrodes on the elevator muscles (masseter and temporalis) during mastication of cold beef. Two different textures (T(1): tough and dry; T(2): tender and juicy) were studied, and subjects were asked to chew the beef and then spit out the bolus either: (1) after a constant chewing period of 7s or (2) when the bolus was ready to be swallowed. Meat samples were weighed before and after chewing to determine weight changes due to saliva incorporation and the release of meat juice. Cutting tests were applied to measure the maximum shear force. The mechanical shear force was maximal for meat before chewing (T(1)=124 N/cm(2); T(2)=83 N/cm(2)) and decreased with increased chewing duration. Texture differences analysed from mechanical measurements remained significant even when the boli were ready for swallowing (T(1)=39 N/cm(2); T(2)=32 N/cm(2)); the toughest meat gave the toughest bolus. Muscular activity adapted to the texture of the meat as soon as chewing began, and remained constant over the observed chewing period. Mean muscular activity was higher during the chewing of tough meat than during the chewing of tender meat. As a consequence, by the time a bolus was ready to be swallowed, more saliva had been incorporated into the tough meat samples (mean weight increase: 36%) than the tender meat samples (mean weight increase: 30%). PMID- 12648557 TI - The effect of chewing bicarbonate-containing gum on salivary flow rate and pH in humans. AB - OBJECTIVE: Gum chewing increases salivary flow rate and pH. The aim of this study was to compare the effects of chewing standard sugar-free gum with those of a gum containing sodium bicarbonate. DESIGN: Whole mouth saliva was collected from 20 volunteers who met inclusion criteria and gave informed consent. After unstimulated saliva was collected, stimulated saliva was collected at intervals during 30 min of chewing either a standard, mint-flavoured gum or bicarbonate containing, mint-flavoured gum. The salivary flow and pH were measured for each sample. RESULTS: With the standard gum, the mean peak salivary flow rate was 3.1+/-1.27 ml/min and the peak salivary pH was 7.39+/-0.14. With the bicarbonate gum, the peak flow rate was 2.79+/-1.38 ml/min and the peak salivary pH was 8.06+/-0.18. The salivary flow rates with the two gums were not significantly different; however, the increase in salivary pH was significantly greater for the bicarbonate gum. CONCLUSION: The increased salivary pH with bicarbonate gum may have implications for oral health and prevention of dental caries. PMID- 12648558 TI - Modulation by somatostatin of rat submandibular salivary secretion. AB - Although somatostatin (somatotrophin release inhibitory factor; SRIF) is a well known inhibitory peptide, there are only a few reports of it acting as a positive modulator. In this work, the action of somatostatin upon rat submandibular protein secretion was studied. In vivo somatostatin infusion (35 microg/(kg h)) raised protein secretion stimulated by adrenergic and peptidergic agents. To rule out possible systemic effects of somatostatin, in vitro experiments were performed. Somatostatin (90 nmol/l) augmented protein release stimulated by noradrenaline (19 micromol/l) and substance P (10 micromol/l), but it did not affect isoprenaline (400 micromol/l)-induced protein release. Phenoxybenzamine (20 micromol/l) reduced the effect of somatostatin on noradrenaline-stimulated protein release. Propranolol (20 micromol/l) increased the noradrenaline stimulated protein release and this effect was synergistic with the action of somatostatin. The absence of extracellular calcium did not significantly reduce somatostatin enhancement of agonist-induced secretion. Fluorescence measurements of the Ca(2+)-sensitive dye fluo3 showed that cytosolic calcium in acinar cells remained elevated during stimuli when somatostatin was present in the medium. It was concluded that somatostatin modulates rat submandibular protein secretion by prolonging the time that the cytosolic calcium signal remains high after stimulus. PMID- 12648559 TI - Evidence for fibroblast growth factor receptors in myofibroblasts during palatal mucoperiosteal repair. AB - Fibroblast growth factors (FGFs) regulate cell growth and differentiation and play crucial roles in the process of tissue repair and remodelling. We have previously shown that basic FGF is widely expressed at the injured site. Since the presence of FGF receptors (FGFRs) determines cellular responsiveness, we examined the localisation of FGFR1, FGFR2 and FGFR3 expression by immunohistochemistry throughout the repair of full-thickness excisional wounds up to 28 days after wounding. Strong expression of FGFR1 was observed in the nuclei of myofibroblasts, which are characterised by alpha-smooth muscle (alpha-SM) actin expression. The weak expression of FGFR2 was also observed in the nuclei of myofibroblasts. In contrast, there was no staining for FGFR3 in fibroblasts through the wound healing process. In addition, transforming growth factor-beta1 (TGF-beta1), a potential inducer of myofibroblasts, enhanced the expression of FGFR1 and FGFR2 in the nuclei of palatal fibroblasts in vitro. These findings suggest that FGFR1 and FGFR2 in myofibroblasts may be responsible for the signal transduction of FGF during the wound healing process. PMID- 12648561 TI - Sialolith crystals localized intraglandularly and in the Wharton's duct of the human submandibular gland: an X-ray diffraction analysis. AB - The exact cause of the formation of sialoliths is unknown. Detailed knowledge of pathogenesis of sialolithiasis and composition of sialoliths is necessary to define new therapeutic procedures. The crystalline components of 23 sialoliths of human submandibular gland were investigated by X-ray powder diffraction analysis. All of the sialoliths localized in the ducts in the submandibular gland consisted of hydroxylapatite. However, in the sialoliths in the Wharton's duct, hydroxylapatite as well as whitlockite and brushite could be found in all except one case. Whitlockite was observed more often in the nucleus of the sialoliths and it was a common co-phase along with hydroxylapatite. The nucleus in one sialolith consisted of brushite and the cortex showed a co-phase of hydroxylapatite and brushite. The occurrence of whitlockite in the sialoliths in Wharton's duct may be due to a higher concentration of calcium and phosphate in saliva in this duct. PMID- 12648560 TI - Biodistribution of a carborane-containing porphyrin as a targeting agent for Boron Neutron Capture Therapy of oral cancer in the hamster cheek pouch. AB - Boron Neutron Capture Therapy (BNCT) is a bimodal cancer treatment based on the selective accumulation of 10B in tumors and concurrent irradiation with thermalized neutrons. The short-range, high-LET radiation produced by the capture of neutrons by 10B could potentially control tumor while sparing normal tissue if the boron compound targets tumor selectively within the treatment volume. In previous studies, we proposed and validated the hamster cheek pouch model of oral cancer for BNCT studies, proved that absolute and relative uptake of the clinically employed boron compound boronophenylalanine (BPA) would be potentially therapeutic in this model and provided evidence of the efficacy of in vivo BPA mediated BNCT to control hamster oral mucosa tumors with virtually no damage to normal tissue. We herein present the biodistribution and pharmacokinetics of a lipophilic, carborane-containing tetraphenylporphyrin (CuTCPH) in the hamster oral cancer model. CuTCPH is a novel, non-toxic compound that may be advantageous in terms of selective and absolute delivery of boron to tumor tissues. For potentially effective BNCT, tumor boron concentrations from a new agent should be greater than 30 ppm and tumor/blood and tumor/normal tissue boron concentration ratios should be greater than 5/1 without causing significant toxicity. We administered CuTCPH intraperitoneally (i.p.) as a single dose of 32 microg/g body weight (b.w.) (10 microg B/g b.w.) or as four doses of 32 microg/g b.w. over 2 days. Blood (Bl) and tissues were sampled at 3, 6, 12, 24, 48, and 72 h in the single-dose protocol and at 1-4 days after the last injection in the multidose protocol. The tissues sampled were tumor (T), precancerous tissue surrounding tumor, normal pouch (N), skin, tongue, cheek and palate mucosa, liver, spleen, parotid gland and brain. The maximum mean B ratios for the single-dose protocol were T/N: 9.2/1 (12h) and T/Bl: 18.1/1 (72 h). The B value peaked to 20.7+/-18.5 ppm in tumor at 24h. The multidose protocol maximum mean ratios were T/N: 11.9/1 (3 days) and T/Bl: 235/1 (4 days). Absolute boron concentration in tumor reached a maximum value of 116 ppm and a mean value of 71.5+/-48.3 ppm at 3 days. The fact that absolute and relative B values markedly exceeded the BNCT therapeutic threshold with no apparent toxicity may confer on this compound a therapeutic advantage. CuTCPH-mediated BNCT would be potentially useful for the treatment of oral cancer in an experimental model. PMID- 12648562 TI - Transmission and scanning electron microscopic observations of epithelial mesenchymal interface of rat tooth germs using dithiothreitol separation. AB - Aperiodic fibrils (AF) project from the interstitial side of the lamina densa of the basement membrane (BM) of the inner enamel epithelium (IE), and show remarkable changes in their morphology during development. The three-dimensional morphology of aperiodic fibrils during development has not been observed, because of the difficulty of exposing the interstitial surface of the BM of the inner enamel epithelium. In the present study, the dithiothreitol separation method was applied to expose the interstitial side of the inner enamel epithelial BM of rat tooth germs for the purpose of observing the exposed aperiodic fibrils by transmission and scanning electron microscopy (TEM and SEM, respectively). After dithiothreitol treatment, the enamel organ (EO) was mechanically separated from the dental papilla (DP). In the region with poorly-developed aperiodic fibrils, the separation occurred at the junction between the inner enamel epithelial BM and the dental papilla, and the aperiodic fibrils were exposed, showing the typical picture of dithiothreitol separation. SEM observation of this region revealed that the aperiodic fibrils were connected to each other and they formed networks. These networks resembled those formed by the anchoring fibrils of epidermal and mucosal epithelial BMs. TEM and SEM observations revealed that there were sidechain-like structures on the surface of the aperiodic fibrils. In the region with well-developed aperiodic fibrils, dithiothreitol treatment was not entirely effective, and some mesenchymal tissues remained on the BM. In this region, TEM observation revealed that the aperiodic fibrils were arranged in parallel with each other, and were connected by the sidechains. Several thin collagen fibrils, which were thought to be immature collagen fibrils (CF) of the predentine, were also connected to the aperiodic fibrils with these sidechains and arranged in parallel with them. Based on SEM and TEM observations, the aperiodic fibrils may be regarded as a kind of anchoring fibrils and they may play a role in connecting the BM with the mesenchymal tissue below. They are also thought to guide the arrangement of collagen fibrils in the surface layer of the predentin. PMID- 12648563 TI - Extraction and radioimmunoassay quantitation of neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) from human dental pulp tissue. AB - The measurement of neuropeptides in complex biological tissue samples requires efficient and appropriate extraction methods so that immunoreactivity is retained for subsequent radioimmunoassay detection. Since neuropeptides differ in their molecular mass, charge and hydrophobicity, no single method will suffice for the optimal extraction of various neuropeptides. In this study, dental pulp tissue was obtained from 30 human non-carious teeth. Of the three different neuropeptide extraction methods employed, boiling in acetic acid in the presence of protease inhibitors yielded the highest levels of neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP). High pressure liquid chromatography (HPLC) analysis of dental pulp tissue verified the authenticity of the neuropeptides extracted. PMID- 12648564 TI - Models of bacteriophage DNA packaging motors. AB - An ATP-dependent motor drives a DNA genome into a bacteriophage capsid during morphogenesis of double-stranded DNA bacteriophages both in vivo and in vitro. The DNA molecule enters the capsid through a channel in the center of a symmetric protein ring called a connector. Mechanisms in two classes have been proposed for this motor: (1) An ATP-driven rotating connector pulls a DNA molecule via serial power strokes. (2) The connector rectifies DNA motion that is either thermal, biased thermal, or oscillating electrical field-induced (motor-ratchet hypothesis). Mechanisms in the first class have previously been proposed to explain the detailed structure of DNA packaging motors. The present study demonstrates that the motor-ratchet hypothesis also explains the current data, including data in the following categories: biochemical genetics, energetics, structure, and packaging dynamics. PMID- 12648565 TI - Structure of human chromosomes studied by atomic force microscopy. Part II. Relationship between structure and cytogenetic bands. AB - In the first part of this work, human chromosomes were characterized by atomic force microscopy (AFM) in air and in aqueous solution. The analysis of the images suggests that the last level of organization consists of a radial arrangement of chromatin loops which are anchored to a fiber which is folded giving a pattern of bands which differs in volume. Here the pattern of bands observed by AFM is compared to the cytogenetic map at the 850-band level. Thus thicker and thinner bands are identified as G and R bands, respectively. Finally a model is proposed which links genome sequence, cytogenetics, and chromosome structure. PMID- 12648566 TI - Structure of human chromosomes studied by atomic force microscopy. AB - In this work human chromosomes have been treated with RNase and pepsin to remove the layer of cellular material that covers the standard preparations on glass slides. This allows characterization of the topography of chromosomes at nanometer scale in air and in physiological solution by atomic force microscopy. Imaging of the dehydrated structure in air indicates radial arrangement of chromatin loops as the last level of DNA packing. However, imaging in liquid reveals a last level of organization consisting of a hierarchy of bands and coils. Additionally force curves between the tip and the chromosome in liquid are consistent with radial chromatin loops. These results and previous electron microscopy studies are analyzed, and a model is proposed for the chromosome structure in which radial loops and helical coils coexist. PMID- 12648567 TI - Characteristics of mineral particles in the human bone/cartilage interface. AB - Bone and cartilage consist of different organic matrices, which can both be mineralized by the deposition of nano-sized calcium phosphate particles. We have studied these mineral particles in the mineralized cartilage layer between bone and different types of cartilage (bone/articular cartilage, bone/intervertebral disk, and bone/growth cartilage) of individuals aged 54 years, 12 years, and 6 months. Quantitative backscattered electron imaging and scanning small-angle X ray scattering at a synchrotron radiation source were combined with light microscopy to determine calcium content, mineral particle size and alignment, and collagen orientation, respectively. Mineralized cartilage revealed a higher calcium content than the adjacent bone (p<0.05 for all samples), whereas the highest values were found in growth cartilage. Surprisingly, we found the mineral platelet width similar for bone and mineralized cartilage, with the exception of the growth cartilage sample. The most striking result, however, was the abrupt change of mineral particle orientation at the interface between the two tissues. While the particles were aligned perpendicular to the interface in cartilage, they were oriented parallel to it in bone, reflecting the morphology of the underlying organic matrices. The tight bonding of mineralized cartilage to bone suggests a mechanical role for the interface of the two elastically different tissues, bone and cartilage. PMID- 12648568 TI - Identification of minimal peptide sequences in the (8-20) domain of human islet amyloid polypeptide involved in fibrillogenesis. AB - We have examined a series of overlapping peptide fragments from the 8-20 region of human islet amyloid polypeptide (IAPP) with the objective of defining the smallest fibril-forming domain. Peptide fragments corresponding to LANFLV (residues 12-17) and FLVHSS (residues 15-20) were strong enhancers of beta-sheet transition and fibril formation. Negative stain electron microscopy illustrated the ability of these peptide fragments to form fibrils independently when incubated alone in solution. Circular dichroism analysis revealed that when full length human IAPP was incubated in the presence of these two fragments, fibrillogenesis was accelerated. While the two fragments, LANFLV and FLVHSS, were able to enhance the recruitment of additional IAPP molecules during fibril formation, the "seeding" activity of these peptides had no effect on altering IAPP-induced cytotoxcity as determined by cell culture studies. Therefore, this study has identified two internal IAPP peptide fragments within the 8-20 domain that may have a role in enhancing the folding and aggregation of human IAPP. These fragments are the smallest sequences identified, within the 8-20 region of hIAPP, that can independently form fibrils, and that can interact with IAPP to assemble into fibrils with characteristics similar as those formed by human IAPP alone. PMID- 12648569 TI - Gold finder: a computer method for fast automatic double gold labeling detection, counting, and color overlay in electron microscopic images. AB - This work presents a computerized method to identify, detect, evaluate, and, by colored overlay, present gold particle pairs in electron microscopy (EM), even in wide-field views. Double gold immunolabeled specimens were analyzed in a LEO 912 electron microscope equipped with a 2k x 2k-pixel slow-scan cooled CCD camera connected to a computer with analySIS 3.1 PRO image processing software. The acquisition of a high-resolution and high-dynamic-range image by the camera allowed correct segmentation of the gold particles, separating them from other cell structures and from the substrate. Particle identification was performed by a classification module designed by us. Based on shape and size, the computer recognized the group of small particles and classified them as either singular or clustered and differentiated these from the single bigger type. The final image shows the particle types separated and colored, and indicates the total number of objects encountered in the specific region of interest. Moreover, a montage tool allowed us to obtain final representative images of large microscopic fields, which on analysis by the Gold Finder module provided information on the distribution and localization of antigens comparable to that provided by the wide field light microscope images. PMID- 12648570 TI - Interaction of GroEL and GroEL/GroES complexes with a nonnative subtilisin variant: a small-angle neutron scattering study. AB - Small-angle neutron scattering and contrast variation were used to study the solution structure of GroEL and GroEL/GroES chaperonins complexed with a nonnative variant of the polypeptide substrate, subtilisin (PJ9). The subtilisin was 86% deuterated (dPJ9) so that it contrasted sufficiently with the chaperonin, allowing the contrast variation technique to be used to separate the scattering from the two components bound in the complex. Both the native double-ring GroEL and a single-ring mutant were used with dPJ9 bound in a 1:1 stoichiometry per GroEL toroid. This allowed both the position and the shape of dPJ9 in the GroEL/dPJ9 complexes to be determined. A single-ring GroEL/GroES variant complexed with one dPJ9 molecule was used to study the structural changes of dPJ9 in GroEL/GroES/dPJ9 complexes formed with ADP and with ATP. It was found that both the shape and the position of the bound dPJ9 in the GroEL/GroES/dPJ9 complex with ADP were the same as those in the GroEL/dPJ9 complex. However, dPJ9 assumed a more symmetric shape when bound in the GroEL/GroES/dPJ9 complex with ATP. This important observation reflects the relative ability of ATP to promote refolding of protein substrates relative to that of ADP. PMID- 12648571 TI - Nuclear pore complexes exceeding eightfold rotational symmetry. AB - Nuclear pore complexes are rotationally symmetric structures that span the nuclear envelope and provide channels for nucleocytoplasmic traffic. These large complexes normally consist of eight spokes arranged around a central channel, although, occasionally, 9- and 10-fold nuclear pore complexes are found in preparations of Xenopus oocyte macronuclei. Here we examine these unusual nuclear pore complexes by negative stain electron microscopy and image analysis and compare the results with data previously obtained from 8-fold structures. The details in two-dimensional and three-dimensional maps indicate that the substructure of the spoke is the same in 8-, 9- and 10-fold nuclear pore complexes: therefore, the spoke is likely an immutable structural component. In all three variant forms, the spacing between adjacent annular subunits, which surround the central channel, is identical. Distances between spokes at higher radius decrease in the 9- and 10-fold nuclear pore complexes. These data imply that the most important connections holding the nuclear pore complex together are those between adjacent annular subunits and that these interactions may play a predominant role in nuclear pore complex assembly. Circumferential connections mediated by ring subunits and radial arms presumably further stabilize the structure and are flexible enough to accommodate additional spokes. PMID- 12648572 TI - HER2/neu overexpression: has the Achilles' heel of uterine serous papillary carcinoma been exposed? PMID- 12648573 TI - Phase II study of gemcitabine in recurrent platinum-and paclitaxel-resistant ovarian cancer. AB - OBJECTIVE: To determine the activity and tolerability of gemcitabine in the palliative treatment of ovarian cancer. METHODS: Patients affected by ovarian cancer, and with progressive disease after treatment with platinum/paclitaxel based chemotherapy, were enrolled into this phase II study. Gemcitabine, 1000 mg/m(2), was administered on days 1, 8, and 15, by 30-min intravenous infusion. Cycles were repeated every 28 days. RESULTS: Fifty patients were enrolled. All the patients were platinum and/or paclitaxel resistant (median number of previous regimens, 2; range, 1-5). Median platinum-free interval was 3 (range, 1-11) months and median paclitaxel-free interval was 6 (range, 1-36) months. A total of 210 courses were evaluable for toxicity, with a median number of four cycles administered per patient (range, 1-10). A grade 3 or 4 hematological toxicity was observed in 27 patients (54%) (anemia grade 3, 16%; grade 4, 2%; neutropenia grade 3, 24%; grade 4, 18%; thrombocytopenia grade 3, 8%; grade 4, 0%). A 20-50% dose reduction was required for 36 patients (72%, 55% of cycles). Blood transfusions were necessary for 15 patients (30%), while 2 (4%) were treated with erythropoetin. Granulocyte colony-stimulating factor was necessary in 4 patients (8%). Nonhematological toxicity was mild and manageable. Only 4 patients (8%) experienced a grade 3 hepatic toxicity (elevated liver enzymes). Forty-one patients (82%) are, so far, evaluable for response. Among them, 7 partial responses (17%; 95% confidence interval [CI], 6-29), 15 disease stabilizations (>16 weeks) (36.6%; 95% CI, 21.9-51.3), and 19 progressions (46.3%; 95% CI, 31.0 61.6) have been registered. An overall clinical benefit was observed in 53.7% of patients. Thirteen patients (31.7%) had a time-to-progression exceeding 24 weeks. CONCLUSIONS: This study confirms the activity and safety of gemcitabine in heavily pretreated patients with recurrent ovarian cancer. PMID- 12648574 TI - Physical side effects and quality of life during postoperative radiotherapy for uterine cancer. Prospective evaluation by a diary card. AB - OBJECTIVE: Adjuvant pelvic radiotherapy (XRT) after radical surgery for uterine cancer may cause various (mainly gastrointestinal) side effects capable of affecting a patient's lifestyle. This prospective study used a diary card to evaluate toxicity and daily quality of life changes in uterine cancer patients given postoperative pelvic radiotherapy. METHODS: The self-completed diary card listed 12 items concerning lifestyle changes and side effects. The patients were stratified by age (65 years). RESULTS: Twenty-five patients were evaluable, and a total of 848 days were assessed (82.6% of the 1027 days of XRT). The mean number of daily stools progressively increased during treatment (P < 0.01). No statistically significant differences were observed in terms of nausea or pain. There was a significant decrease in appetite (P < 0.01). Vaginal symptom scores did not significantly change during treatment, and no significant differences were found in terms of anxiety, daily activities, or compliance to therapy. The subjective perception of overall well-being and the quality of life score progressively and significantly decreased over time (P < 0.01). The younger patients had significantly more bowel movements, less appetite, more pain and vaginal burning, less vaginal bleeding, more anxiety, a greater reduction in daily activities and therapeutic compliance, a poorer subjective perception of overall well-being, and a lower quality of life score. CONCLUSIONS: The use of a diary card may be an adequate means of detecting the extent of treatment-related changes in the lifestyle of uterine cancer patients receiving postoperative radiotherapy. PMID- 12648575 TI - Activity of paclitaxel as second-line chemotherapy in endometrial carcinoma: a Gynecologic Oncology Group study. AB - OBJECTIVE: To estimate the antitumor activity of paclitaxel (Taxol) in patients with persistent or recurrent endometrial carcinoma who have failed prior chemotherapy. To determine the nature and degree of toxicity of paclitaxel in this group of patients. METHODS: Paclitaxel was administered as a 3-h infusion at an initial dose of 200 mg/m(2) every 21 days or 175 mg/m(2) for patients with prior pelvic radiation therapy. Dose modifications were based on nadir toxicity, both hematologic and nonhematologic, and were accomplished by dose level adjustments. The dose levels were 200, 175, 135, and 110 mg/m(2). Patients were evaluable for response after receiving one dose of paclitaxel and living 3 weeks. They were evaluable for toxicity after receiving any paclitaxel. RESULTS: Of the 44 patients evaluable for response, three patients (6.8%) achieved a complete response and nine patients (20.5%) had a partial response for an overall response rate of 27.3%. The 95% confidence interval for the true response rate was 15 42.8%. The median number of courses of paclitaxel to response was 2 (range: 1-4) and the median response duration was 4.2 months. The median overall survival was 10.3 months. Of 48 patients evaluable for toxicity, 28 experienced at least one episode of grade 3 or 4 neutropenia, with one treatment-related death. There were four patients who developed grade 3 neurotoxicity in this group of previously treated patients, most of whom had received cisplatin-containing chemotherapy. There was virtually no cardiac toxicity and only 3 of 48 patients experienced grade 3 or 4 gastrointestinal symptoms. CONCLUSIONS: Paclitaxel is an active agent in the treatment of endometrial cancer in patients who have had prior chemotherapy. PMID- 12648576 TI - Phase II trial of single agent carboplatin followed by dose-intense paclitaxel, followed by maintenance paclitaxel therapy in stage IV ovarian, fallopian tube, and peritoneal cancers: a Southwest Oncology Group trial. AB - OBJECTIVE: To improve the survival of women with stage IV epithelial ovarian cancer the Southwest Oncology Group conducted a phase II trial examining two novel treatment strategies: (a) modification of the chemotherapy regimen (carboplatin/paclitaxel) based on evidence of response during treatment and (b) "maintenance therapy," with the monthly administration of single agent paclitaxel (maximum 2 years). METHODS: Individuals with histologically documented stage IV ovarian, fallopian tube, or peritoneal cancers initially received a maximum of 3 courses of single agent carboplatin (AUC 6), with the decision to administer a subsequent course based on specific response criteria (i.e., declines in serum CA 125 or shrinkage of measurable mass lesions). This was followed by single agent paclitaxel (150 mg/m(2)), delivered every 2 weeks for a maximum of 6 courses, again based on evidence of continued response to the treatment program. Finally, in the absence of disease progression or unacceptable toxicity, patients received single agent paclitaxel (175 mg/m2) for a maximum of 24 monthly courses. The primary study end point was 2-year overall survival, which was to be compared to the survival of a previously published historical control population [stage IV ovarian cancer patients treated with a "standard" cisplatin/paclitaxel regimen (GOG 111; McGuire WP, N Eng E J Med 1996; 334:1)]. This analysis was employed to determine if this novel strategy was worthy of further clinical investigation in a controlled clinical trial. RESULTS: There were 53 patients entered this multicenter study, of whom 50 were evaluable for toxicity and 51 for survival. There were 2 treatment-related deaths. Grades 2 and 3 neutrotoxicity were observed in 18 and 4% of patients, respectively. There were no major treatment related protocol violations. The overall 2-year survival was 48% (95% CI, 34 62%), compared to 60% for the historical control group. CONCLUSIONS: While this trial demonstrated the feasibility of delivering a treatment regimen requiring frequent alternations in therapy based on predetermined changes in objective parameters of response, further exploration of this specific management approach does not appear warranted in stage IV ovarian cancer, based on the survival outcome compared to a reasonably similar historical control population. As this trial demonstrates, the prospective determination of clinical end points in a phase II study (e.g., 2-year survival) that will lead to continued evaluation of a particular investigative strategy in a large-scale randomized trial can be an effective method to reduce the bias and uncertainty often associated with this complex decision process. PMID- 12648577 TI - Immunohistochemical and quantitative competitive PCR analyses of midkine and pleiotrophin expression in cervical cancer. AB - OBJECTIVE: The aim of this study was to determine midkine (MK) and pleiotrophin (PTN) expression in cervical cancer. METHODS: Prospective study in tertiary teaching hospital. Normal and cancerous cervical tissues were obtained from healthy women (n = 19) and from patients with cervical cancer (n = 42). The expressions of MK and PTN mRNA and protein were examined by quantitative competitive PCR and by immunohistochemistry. MK and PTN mRNA and protein expressions were examined with respect to tumor stage and size. RESULTS: The expressions of midkine and pleiotrophin mRNA in cervical cancer were higher than those in the normal cervix (MK, 175.59 +/- 63.3 vs 1.00 +/- 0.18 fmol, respectively; PTN, 3.18 +/- 1.25 vs. 0.86 +/- 0.12 fmol, respectively, P < 0.05), and their expressions were not correlated with cervical cancer stage or size of the tumor. The expressions of MK and PTN protein in cancerous tissue were higher than those in the normal cervix (P < 0.05). Moreover, the protein expression of MK, but not of PTN, correlated with tumor stage and size. The expressions of MK and PTN were not correlated with vascular density. CONCLUSIONS: Our results suggest that increased midkine mRNA and protein expressions are associated with the carcinogenesis of cervical cancer. PMID- 12648578 TI - Intratumoral color Doppler analysis in endometrial carcinoma: is it clinically useful? AB - OBJECTIVE: The aim was to study the effectiveness of subjective color Doppler evaluation and spectral Doppler parameters in preoperative characterization of endometrial carcinomas. METHODS: Seventy-six patients with endometrial carcinoma were preoperatively analyzed by color Doppler ultrasound in order to subjectively evaluate the amount of intratumoral blood flow (color score) and to analyze the lowest resistance index (RI), the highest peak systolic velocity (PV), and the highest time averaged maximum velocity (TAMVX). These parameters were analyzed according to clinico-pathological characteristics. RESULTS: In 13 patients no intratumoral arterial vessels were detected by color Doppler examination. No lymph node metastases were found in this group of patients. Positive nodes were found in 24% of patients with detectable arterial vessels, although the difference did not reach the statistical significance. No differences were found in spectral Doppler parameters (RI, PV, TAMVX) according to tumor characteristics or nodal involvement. A higher percentage of cases with a color score of 3 was found in stage >I than in stage I patients (69 vs 42%, P < 0.05), and in patients with myometrial invasion greater than 50% than in those with less than 50% invasion (72 vs 38%; P = 0.05). CONCLUSIONS: Nodal metastases were found in 24% of patients with detectable vessels at color Doppler examination. Subjective analysis of vessel density correlated >50%, myometrial invasion, but spectral Doppler analysis was not predictive of surgical stage, tumor grade, myometrial invasion, or lymph node metastases. These results do not support the use of preoperative intratumoral blood flow analysis as a clinical test in evaluating tumor characteristics or in predicting lymph node metastases. PMID- 12648579 TI - Prevention of chemotherapy-induced intraperitoneal adhesion formation in rats by icodextrin at a range of concentrations. AB - OBJECTIVE: Two controlled in vivo studies in rats have investigated the effect of icodextrin solution on intraperitoneal chemotherapy-induced adhesion formation. The first study evaluated the effect of three concentrations of icodextrin (4, 15, 20% w/v) in comparison to a phosphate-buffered saline (PBS) control in response to intraperitoneal doxorubicin (n = 40). The second study compared the effect of 4% icodextrin to Ringers' lactate solution (RLS) control in response to intraperitoneal bleomycin (n = 30). METHODS: Doxorubicin and bleomycin were administered via a continuous pump and as a single bolus (bleomycin only). Doxorubicin 2 ml (23.2 microg/ml) was delivered via pump in conjunction with 20 ml of 4, 15, or 20% icodextrin or PBS (n = 10 per group). In the bleomycin experiments rats received either 2 ml (0.77 U/ml) bleomycin delivered via pump in conjunction with 15 ml 4% icodextrin or RLS, or 0.77 or 0.077 U bleomycin delivered in 15 ml 4% icodextrin or RLS administered as a bolus injection (n = 5 per group). Seven days after the initiation of doxorubicin treatment and 9 days after initiation of bleomycin treatment, the rats were euthanized by CO(2) and the extent of peritoneal adhesion formation was evaluated using an 8-point scoring system. RESULTS: When icodextrin was administered in conjunction with doxorubicin there was a reduction in the formation of adhesions compared to PBS. Efficacy increased with the concentration of icodextrin used. The lowest dose of bleomycin (0.077 U) caused very few adhesions. Results with bleomycin 0.77 U/ml (pump) and 0.77 U (bolus) showed that 4% icodextrin was significantly more effective than RLS at preventing adhesion formation, irrespective of the dosing regimen. CONCLUSIONS: These studies suggest that 4% icodextrin may reduce adhesion formation caused by intraperitoneal chemotherapy. PMID- 12648580 TI - Toward understanding the natural history of ovarian carcinoma development: a clinicopathological approach. AB - OBJECTIVE: The natural history of the development of ovarian carcinoma is not known. It also remains undetermined whether ovarian carcinomas develop from benign and/or borderline malignant tumors or arise de novo from the ovarian surface epithelium. METHODS: To address these issues clinicopathologically, we reviewed the clinical charts of 543 patients with epithelial ovarian carcinoma and 252 patients with borderline tumors who underwent laparotomy at seven hospitals and collected patients whose clinical and transvaginal ultrasonography (USG) findings for adnexal regions 12 months or fewer prior to the surgery were available. Histological slides of the resected specimens were reexamined concerning the diagnosis and histological grade, as well as the presence or absence of benign- or borderline-like lesions adjacent to the carcinoma. RESULTS: Forty-nine patients had had gynecological examination with transvaginal USG 12 months or fewer prior to laparotomy. Among them, 35 had carcinomas (11 serous, 6 mucinous, 8 clear cell, 10 endometrioid) and 14 had borderline tumors (8 serous, 6 mucinous). Of the 35 patients with carcinoma, 19 (54%) had been followed up for benign-appearing cysts or endometriotic cysts. In these cases, serial USG examinations revealed an increase in size and/or appearance of the solid part of the cyst. In the remaining 16 (46%), however, there had been no apparent abnormalities in USG, and such cases occurred most frequently for serous carcinomas. CONCLUSIONS: Our findings suggest that approximately half of ovarian carcinomas develop secondarily from preexisting, benign-appearing cysts or endometriotic cysts, whereas the remaining half seem to develop suddenly from a normal-appearing ovary. This appears to be consistent with two possible pathways of ovarian carcinoma development; adenoma-carcinoma sequence and de novo carcinogenesis. PMID- 12648581 TI - p53 mutations and overexpression affect prognosis of ovarian endometrioid cancer but not clear cell cancer. AB - OBJECTIVE: Although ovarian clear cell adenocarcinoma (OCCA) and ovarian endometrioid adenocarcinoma (EC) are considered to be closely related to endometriosis, the mechanisms of carcinogenesis of these two malignancies and malignant transformation of endometriosis are unclear. In this study, we examined the biology of OCCA and EC by performing large-scale analysis of K-ras activation and p53 mutation and overexpression in these malignancies. The results were subsequently analyzed for correlation with the clinicopathologic data. METHODS: In the present study of OCCA and EC, we obtained clinicopathological data and analyzed frequency of mutations and overexpression of K-ras and p53. DNA was extracted from formalin-fixed, paraffin-embedded tissue, and target sequences were amplified in vitro by polymerase chain reaction. The DNA was analyzed for K ras and p53 mutations by testing for single-strand conformation polymorphisms and by direct sequencing. Immunohistochemical staining was performed using p53 monoclonal antibody. Univariate analysis was performed using the Kaplan-Meier algorithm, and differences in survival were analyzed using the log rank test. The prognostic significance of the studied variables for survival was assessed using multivariate analysis with Cox regression analysis. RESULTS: K-ras mutation was detected in 16.2% (6/37) of OCCA patients and 3.7% (1/27) of EC patients. No evidence of p53 mutation was detected in OCCA patients, but p53 mutation was detected in 63.0% of EC patients; these findings are consistent with the results of p53 immunohistochemistry. No statistical significance was observed for K-ras mutation in OCCA or EC. In EC patients, the absence of endometriosis and p53 overexpression was associated with a poorer survival. In OCCA patients tubulocystic and papillary histotype as well as stage II correlated with a worse survival. CONCLUSIONS: p53 mutation, which was found in 63% of EC tumors, is an independent prognostic factor for EC patients. However, no p53 mutation was found in OCCA tumors. K-ras mutations did not affect survival of OCCA or EC patients. PMID- 12648582 TI - TAP1, TAP2, and HLA-DR2 alleles are predictors of cervical cancer risk. AB - OBJECTIVE: The likelihood of developing cervical cancer has been shown to be increased in persons with certain HLA alleles. We evaluated immune response genes in the HLA region of chromosome 6 to see if individual or interactive associations with cervical cancer risk could be identified. METHODS: Tissue was obtained from 127 women undergoing surgical treatment for cervical cancer. Blood samples were obtained from 175 control subjects. A combination of polymerase chain reaction (PCR), sequence-specific PCR, and DNA sequencing was used to evaluate polymorphic alleles, including HLA class I B7, TNF alpha, HLA class II DR2, TAP1, and TAP2 genes. Fisher's exact test and logistic regression modeling were used for statistical analysis. RESULTS: A significantly greater proportion of the patients with cervical cancer were found to have the HLA class II DR2 1501 allele (P = 0.023) and the TAP2 A/B heterozygous pattern of alleles (P = 0.0006) than were women without cervical cancer. A proportion of patients with cervical cancer significantly smaller than that of the control women had a polymorphism at the -238 position of the TNF promoter and the TAP1 C/C homozygous pattern of alleles. With logistic modeling, the markers that showed consistent association with the occurrence of cervical cancer were TAP2 A/B, HLA-DR2 1501, and TAP1 C/C. CONCLUSIONS: We demonstrated a significant association between immune response genes and the risk of cervical cancer. Our data create a compelling argument for a gene or a cluster of genes in the HLA region of chromosome 6 that regulates host immune responses to human papillomavirus infection in a manner that results in inherited susceptibility or resistance to the transforming properties of oncogenic papillomaviruses. PMID- 12648583 TI - Endocrine tumors of the uterine cervix: incidence, demographics, and survival with comparison to squamous cell carcinoma. AB - OBJECTIVE: The aim was to describe the epidemiology of endocrine tumors of the cervix in comparison with invasive squamous cell carcinomas using population based data reported to the Surveillance, Epidemiology and End-Results (SEER) program. METHODS: Retrospective analysis of actively followed cases reported to SEER from 1973 to 1998. Incidence, demographic characteristics, and survival were compared for endocrine and squamous tumors. RESULTS: There were 239 cases of endocrine tumors and 18,458 cases of invasive squamous cell carcinoma of the cervix included in the study. Mean age at diagnosis was 49 years for endocrine tumors versus 52 years for squamous cell carcinoma (P < 0.01). Endocrine tumors were more likely to present at a later FIGO stage (P < 0.01), and to have lymph node involvement at diagnosis (57 vs 18%, P < 0.01) compared to squamous cell carcinoma. Observed median survival for women with endocrine tumors was 22 months versus 10 years for women with squamous cell carcinoma. Age and FIGO stage adjusted hazards of death were 1.84 times greater for endocrine tumors than for squamous cell carcinoma (95% CI 1.52-2.23). At all stages of disease, survival was worse for women with endocrine tumors compared to women with squamous cell carcinomas. CONCLUSIONS: Endocrine tumors of the cervix are extremely aggressive and survival is poor regardless of stage at diagnosis. PMID- 12648584 TI - Histologic findings from the cervix among older women with abnormal cervical cytology. AB - OBJECTIVE: The aim of this study was to determine rates of cervical neoplasia among women at least 50 years of age referred for colposcopy after abnormal cytology and to compare these to younger women. METHODS: From a prospectively accrued database of 2825 women undergoing colposcopy in the gynecology clinic of an urban public hospital, women at least 50 years of age with a known cytologic abnormality were selected for retrospective analysis. Demographic and medical information collected at colposcopy and subsequent histology was reviewed. Cytology results were based on the Bethesda system, and histology was reported as grades of cervical intraepithelial neoplasia (CIN). Statistical analysis was by t test, chi(2) test, and Mann-Whitney U test. RESULTS: Among 325 women at least age 50, cervical histologic results were benign or atypical for 147 (45%), CIN1 for 28 (9%), CIN2 for 21 (6%), CIN3 for 49 (15%), cancer for 11 (3%), and ungraded dysplasia for 7 (2%), with no biopsy performed for 62 (19%) women. Symptoms were more common among women with cancer (6/11 or 55%) than those without (62/263 or 21%, P = 0.01). Negative histology (80/231 or 35%), CIN3 (49/231 or 21%), and cancer (9/231 or 4%) were more common among older than younger women (287/1403 or 20%, 199/1403 or 14%, and 11/1403 or 1%, respectively), while atypia (438/1403 or 31%), CIN1 (321/1403 or 23%), and CIN2 (147/1403 or 10%) were more common among younger than older women (53/231 or 23%, 23/231 or 10%, and 17/231 or 7%, respectively, P < 0.04). Differences in the distribution of cervical histology results remained significant among women with ASCUS (P = 0.001) but not those with LSIL (P > 0.9), HSIL (P > 0.07), or cancer (P > 0.4). CONCLUSIONS: Most older women are referred for colposcopy with lesser grades of abnormality, but cervical cancers are found across all cytologic grades and were more common in symptomatic women. Compared to younger women with abnormal cytology, women at least 50 years of age with ASCUS had higher rates of negative evaluations and high grade but not low- or mid-grade lesions. PMID- 12648585 TI - Human papilloma virus testing in patient follow-up post cone biopsy due to high grade cervical intraepithelial neoplasia. AB - OBJECTIVE: We evaluated the contribution of the human papilloma virus (HPV) load in planning follow-up and management of women post cone biopsy for high-grade cervical intraepithelial neoplasia (CIN2-3). METHODS: Ninety-six suitable women were followed-up by Pap smears: two consecutive abnormal smears dictated referral for colposcopy-directed biopsy. Before colposcopy, HPV tests determined high-risk HPV DNA type and load (Hybrid Capture System type I). Patients histologically diagnosed with CIN1 or CIN2-3 underwent repeat conization or hysterectomy for residual disease. HPV load was compared to cytology for the detection of residual disease. RESULTS: At follow-up, 20/89 (22.4%) studied women had positive cytology reports of either low- (n = 11) or high-grade (n = 9) squamous intraepithelial lesion (SIL). Colposcopic biopsies diagnosed 9 CIN1 and 8 CIN2-3 cases. Residual disease was corroborated in 16/17 (94.1%) women and the status was readjusted based on cone biopsy/hysterectomy: CIN2-3 in 9 and CIN1 in 7. The positive prediction values for CIN2-3 residual disease with high-grade SIL, CIN2-3 on colposcopic punch biopsy, and high HPV load were 89, 100, and 100%, respectively. For CIN1 residual disease with low-grade SIL, CIN1 on colposcopic punch biopsy, and low and borderline HPV load, they were 54.5, 77.7, and 100%. The HPV load was a more accurate predictor for CIN1 or CIN2-3 on the cervical specimen in cases with low-grade SIL or CIN1 on colposcopic biopsy. CONCLUSIONS: Evaluating HPV loads after a positive cytology report may assist in triaging women post conization biopsy for CIN2-3 to appropriate treatment. Its high positive predictive value, specificity, and sensitivity for CIN1 and CIN2-3 and supplementary information could be especially pertinent for clinical management of low-grade SIL cases. PMID- 12648586 TI - Epirubicin and paclitaxel (EPI-TAX regimen) for advanced ovarian cancer after failure of platinum-containing regimens. AB - BACKGROUND: The place of anthracyclines in the treatment of advanced ovarian cancer remains a matter of debate. We have assessed the feasibility and evaluated the tolerance of epirubicin (EPI) combined with paclitaxel (TAX) in heavily pretreated ovarian cancer patients. METHODS: Between March 1996 and March 1998, 34 patients with ovarian cancer in relapse after platinum-based chemotherapy received EPI (75 mg/m(2)/day, iv) and TAX (175 mg/m(2)/day, 3-h infusion). Cycles were repeated every 3 weeks. This treatment was second-line for 10 patients and third/fourth-line for 24. RESULTS: Of the 34 assessable patients, 15 (44%) (95% confidence interval 27-60%) achieved objective response (3 complete and 12 partial responses). The number of previous lines of chemotherapy or previous anthracycline treatments did not influence response rates. Responders to previous paclitaxel-based regimens had a significantly higher response rate to EPI-TAX combination (57%) than nonresponders (11%) (P = 0.05). Median response duration was 40 weeks (range 12-94). Median survival from inclusion was 10.7 months (range 1-40). Myelosuppression was the most frequent side effect. Grade 3/4 neutropenia occurred in 31 patients (91%), febrile neutropenia episodes in 17%, and grade 3/4 anemia and thrombopenia in 27 and 24%, respectively. The main nonhematological toxicities included alopecia and grade 2 peripheral neuropathy (12%). Cardiac dysfunction was observed in one patient after the fourth treatment cycle. CONCLUSIONS: Toxicity of the EPI-TAX regimen was acceptable in this population of heavily pretreated ovarian cancer patients. The regimen was effective and it is considered an option for patients previously responding to paclitaxel-based therapy. PMID- 12648587 TI - Discrepant expression of carboxy- and aminoterminal propeptides of type I procollagen predicts poor clinical outcome in epithelial ovarian cancer. AB - OBJECTIVE: The metabolism and prognostic value of serum concentrations of the aminoterminal (PINP) and carboxyterminal (PICP) propeptides of type I collagen and the PICP:PINP ratio in relation to the carboxyterminal telopeptide of type I collagen (ICTP) in ovarian cancer were evaluated. METHODS: Fifty patients with epithelial ovarian cancer were evaluated with serial measurements of serum concentrations of PICP, PINP, and ICTP before the operation and at 3-month intervals during the first year after the operation. For statistical analysis the patients were divided into two groups according to clinical outcome (alive vs dead) and clinical behavior (fast progression vs others). RESULTS: The serum PINP concentration before the operation was increased and the PICP/PINP ratio was lowered in patients with poor prognosis (PP) compared to those with good prognosis (GP) and in patients with fast progression compared to others. The serum PICP concentration did not differ between the groups. The circulating ICTP concentration was significantly higher in the PP-group than in the GP group. In Kaplan-Meier analysis the PICP:PINP ratio divided the PP and GP patients (P = 0.0004). In multivariate regression analysis, the independent prognostic variables were clinical stage (P = 0.014, 95% confidence interval (CI) 1.31 11.19) and preoperative serum ICTP concentration (P = 0.048, CI 1.01-5.91). When serum ICTP concentration was excluded from the equation, the PICP:PINP ratio (P = 0.012, CI 1.29-7.83), together with clinical stage (P = 0.013, CI 1.31-10.37), was found to be an independent prognostic variable. When the early and advanced stage patients were analyzed separately, the PICP:PINP ratio was a significant prognostic variable in multivariate analysis in early stage patients and in advanced stages during the first 4 years of follow-up. CONCLUSIONS: A low PICP:PINP ratio was associated with fast progression and poor clinical outcome in ovarian cancer. Evaluation of the PICP:PINP ratio together with ICTP may be valuable in predicting the clinical outcome of ovarian cancer. PMID- 12648588 TI - beta-catenin nuclear localization is associated with grade in ovarian serous carcinoma. AB - OBJECTIVE: beta-Catenin has been previously associated with oncogenic activity in human cancers. We evaluated whether beta-catenin also plays a role in papillary serous ovarian neoplasms. METHODS: Immunohistochemistry for beta-catenin was performed on the primary ovarian serous neoplasms of 105 women. Of these, 10 were low malignant potential (LMP) serous tumors, and 95 were serous cancers. Nuclear beta-catenin staining was correlated with grade of tumor and median survival. OVCAR-3, OVCA-420, OVCA-432, and MDAH-277-10c were evaluated for beta-catenin localization and transfected with a T-cell factor (TCF) responsive reporter to evaluate beta-catenin transcriptional activity. RESULTS: Of 105 serous tumors, 13 (12.3%) demonstrated beta-catenin nuclear staining. Eleven of 48 high-grade serous carcinomas (23.0%) demonstrated nuclear staining compared with 1 low-grade serous carcinoma (2.1%) (P = 0.006). One LMP tumor had nuclear staining. beta Catenin nuclear localization was undetectable in the cell lines tested. Furthermore, transient transfection of the cell lines with a TCF-responsive reporter did not demonstrate significant constitutive transcriptional activation. CONCLUSIONS: We found a statistically significant correlation between beta catenin nuclear localization and ovarian high-grade serous carcinomas. Thus, deregulation of beta-catenin may play a role in the pathogenesis of ovarian high grade serous carcinomas in contrast to ovarian low-grade serous carcinomas and LMP serous tumors. PMID- 12648589 TI - Reduced retinoblastoma gene protein to Ki-67 ratio is an adverse prognostic indicator for ovarian adenocarcinoma patients. AB - OBJECTIVE: Alterations in the retinoblastoma gene (RB-1) are common in human neoplasia. However, the clinical significance of the deregulated expression of RB 1 in ovarian cancer remains undefined. We therefore conducted a retrospective investigation to clarify the relationships of RB-1 gene protein (pRb) to the percentage of cycling cells, clinicopathologic variables, other G1 interacting proteins and prognosis of nonbenign epithelial ovarian tumors. METHODS: Paraffin embedded tissue from 127 nonbenign epithelial ovarian tumors, including 44 of low malignant potential (LMP) and 83 primary ovarian adenocarcinomas, was stained immunohistochemically for pRb, p21(Cip1), p27(Kip1), p53, and Ki-67 antigen (a cell proliferation associated marker). Expression of these markers was correlated with clinicopathologic features and with overall survival of patients with adenocarcinomas. RESULTS: pRb levels were significantly lower in LMP tumors than in carcinomas (P = 0.027). In the latter group, pRb expression decreased with increasing grade (I-II vs III) (P = 0.010), advancing stage (I-II vs III) (P < 0.001), and bulk residual disease (P = 0.014). pRb was not related to Ki-67 expression (P > 0.10) or to overall survival (P > 0.10) but a low pRb to Ki-67 ratio emerged as an important indicator of poor survival in univariate analysis in the entire cohort (P = 0.0076) and in the platinum-treated patients (P = 0.0162) as well as in multivariate analysis, along with histologic type and FIGO stage. CONCLUSIONS: Diminished pRb levels are related to several clinicopathologic indicators of aggressiveness in ovarian adenocarcinomas. More importantly, pRb expression coupled with the percentage of Ki-67 positive cells is a better prognostic marker than pRb, Ki-67, or other G1 interacting proteins and supplements the information gained from traditional prognosticators. PMID- 12648591 TI - Endometrial hyperplasia with secretory changes. AB - OBJECTIVE: Secretory changes in endometrial hyperplasia are uncommon. The aim of this study is to review the morphologic and clinical findings of 24 cases of endometrial hyperplasia with secretory changes. METHODS: In 24 patients diagnosed with endometrial hyperplasia with secretory changes during 6 years, clinical characteristics such as menopausal status and hormone treatment were correlated with morphological features. A matched age control group of 24 women with conventional endometrial hyperplasia was used to compare the hormonal effect. RESULTS: Nineteen patients were premenopausal. Nine women showed simple hyperplasia without atypia and 15 complex hyperplasia, 7 of them with atypia. Seventeen women were under hormonal treatment at the time of diagnosis, 10 of them with progestins. In 7 patients endometrial adenocarcinoma could be seen, 5 coexisting with endometrial hyperplasia with secretory changes and in 2 appearing after 1 and 4 months. In control group only 2 patients were undergoing progestin hormonal treatment. CONCLUSIONS: Secretory changes can be found in hyperplastic endometrium, particularly in premenopausal women under hormonal treatment with progestins, with the risk of misdiagnosis. PMID- 12648590 TI - High cyclooxygenase-2 expression in cervical adenocarcinomas. AB - OBJECTIVE: The purpose of this study was to examine the relationships between cyclooxygenase-2 (COX-2) expression and prognostic factors in cervical carcinomas. METHODS: We studied COX-2 expression in 53 women with cervical cancers, including 35 squamous cell carcinomas (SCCs), 1 adenosquamous cell carcinoma (ASCC), and 17 adenocarcinomas (ACs), using commercially available polyclonal antibodies on Formalin-fixed, paraffin-embedded tissues. Normal cervical tissues were obtained as from other patients with uterine myomas treated with a total hysterectomy (n = 16). The immunoreactivity was quantified using an immunohistochemical scoring system that approximates the use of an image analysis based system. RESULTS: Twenty-two cervical cancer tissues (41.5%), including 10 SCCs and 12 ACs, expressed COX-2 at a moderate to strong level, which significantly, differed from the negligible expression found in the control group of 16 normal cervical tissues (P = 0.001). Different cell types showed significantly different expression levels of COX-2 (SCC at 28.6% vs AC at 70.6%, P = 0.004). The presence of deep stromal invasion (n = 40) showed a significant inverse relationship to COX-2 expression (32.5% vs 69.2%, P = 0.02). The expression of COX-2 in well-differentiated carcinomas was significantly increased compared to that in moderately and poorly differentiated carcinomas (72.7% vs 33.3%, respectively, P = 0.018). CONCLUSIONS: Overexpression of COX-2 was found in both SCC and AC, but SCCs showed infrequent and low expression. These findings suggest that increased COX-2 expression may play an important role in cervical adenocarcinomas. PMID- 12648592 TI - Evaluation of complete surgical staging with pelvic and para-aortic lymphadenectomy and paclitaxel plus carboplatin chemotherapy for improvement of survival in stage I ovarian clear cell carcinoma. AB - OBJECTIVE: The aim was to determine the benefits of lymphadenectomy and paclitaxel plus carboplatin chemotherapy for stage I ovarian clear cell carcinoma (defined as intra-abdominal disease confined to the ovaries). METHODS: Twenty patients with stage I pure clear cell carcinoma of the ovary diagnosed between 1991 and 2001 were divided into two groups: Group A (12 patients, 1997-2001) underwent complete surgical staging including bilateral salpingo-oophorectomy, hysterectomy, omentectomy, and pelvic and para-aortic lymphadenectomy, followed by paclitaxel and carboplatin chemotherapy. Group B (8 patients, 1991-1996) underwent bilateral salpingo-oophorectomy, hysterectomy, and omentectomy without lymphadenectomy, followed by cisplatin-based chemotherapy. The survival of the two groups was compared. The clinical characteristics of the two groups were evaluated for age distribution, grade, substage, preoperative CA-125, presence or absence of endometriosis, and maximal tumor diameter. RESULTS: The estimated 4 year survival rate was 76.9%. The clinical characteristics of the two groups were similar, except for lymphadenectomy and regimen of chemotherapy. With a median follow-up of 36 months (range: 11-130 months), one of 12 patients in Group A had recurrence in comparison with 6 of 8 patients in Group B (P = 0.004). The estimated 3-year recurrence-free survival and 4-year overall survival for Group A was significantly greater than that for Group B (91.7 vs 33.3%, P = 0.014; 100 vs 50%, P = 0.014). Median time to recurrence was 8 months. CONCLUSIONS: Complete surgical staging, including pelvic and para-aortic lymphadenectomy and paclitaxel plus carboplatin chemotherapy, appeared to be capable of improving survival of patients with stage I ovarian clear cell carcinoma. PMID- 12648593 TI - Clinical parameters and treatment results in recurrent granulosa cell tumor of the ovary. AB - OBJECTIVE: The objectives of this study were to evaluate the patients' characteristics and treatment results for 11 recurrent granulosa cell tumors (GCT) of the ovary. METHODS: Tumor registries were screened retrospectively for all patients treated between 1979 and 1999 for ovarian tumors at the University of Istanbul. All information about these patients was obtained from operation, pathology, and follow-up reports. Then the clinical data were extracted. The patients were classified in stages according to FIGO criteria. There were 952 ovarian carcinomas, of which 45 were GCT. Eleven of 45 patients (24%) developed recurrent disease on follow-up. RESULTS: All patients but one had advanced (III IV) disease at diagnosis. The median progression-free interval (PFI) was 16 months (range, 5-39 months) with a marked difference in PFI between patients receiving (24 months) and not receiving (8 months) initial chemotherapy. Among patients received chemotherapy (six patients), two complete and two partial responses were observed, for an overall response rate of 67%. The median survival for all patients was 26 (4-73) months with a longer survival (38 versus 8 months) for patients retreated with a platinum-containing regimen. CONCLUSIONS: Despite the fact that small number of treated cases does not allow an adequate evaluation, retreatment with cis-platinum-containing regimens may be the treatment of choice for patients with recurrent disease. PMID- 12648594 TI - Adequate staging for uterine cancer can be performed through Pfannenstiel incisions. AB - OBJECTIVE: To determine if the type of operative incision influences the adequacy of surgical staging in patients with uterine cancer. METHODS: All patients with uterine cancer referred to the Swedish Medical Center Cancer Institute for adjuvant radiotherapy between June 1, 1989, and June 1, 1999, who underwent comprehensive surgical staging and for whom complete records could be obtained were eligible. Data on type of incision, weight, medical comorbidities, histology, total number and distribution of lymph nodes (LN), estimated blood loss, complications, and length of stay were abstracted retrospectively. Statistical analysis with two-tailed Student t test, chi(2), Fisher's exact, and Kaplan-Meier survival curves were performed. RESULTS: Five hundred four women with uterine cancers were referred to the Cancer Institute with 332 meeting inclusion criteria. A vertical midline incision (ML) was used in 236 (72%) while 96 (28%) received a Pfannenstiel incision (PI). No panniculectomies were performed. There were no statistically significant differences in age, weight, stage, histology, comorbidities, or estimated blood loss between the ML and PI groups. ML was associated with significantly more intraoperative and postoperative complications (34 vs. 7; P = 0.003). When compared to ML a greater number of total LN (21.0 vs. 16.8; P = 0.001) and a comparable number of pelvic LN (13.7 vs. 12.2; P = 0.14) were procured through a PI. More patients with a ML (72% vs. 63%; P = 0.13) had para-aortic lymph nodes (PALN) dissected; however, when obtained equivalent numbers of nodes were removed (3.52 vs. 4.36; P = 0.14). Overall, the median length of stay was statistically shorter for those patients operated on via a PI (4 vs. 3 days; P = 0.007). The projected 5-year disease-free (83% vs. 85%) and disease-specific (87% vs. 85%) survival was unaffected by incision. In the heaviest quartile of patients (>180 lb), a statistically greater number of total LN (23.3 vs. 16.5; P = 0.005) and pelvic LN (16.7 vs. 11.5; P = 0.05) were obtained with a PI. Again, PALN were sampled more frequently (67% vs. 56%; P = 0.45) in patients with a ML, but the mean LN yield was no different (3.91 vs. 5.20; P = 0.37). Likewise, in this heaviest quartile, there were no statically significant differences in operative complications (7 vs. 1; P = 0.43) with either incision. CONCLUSIONS: Comprehensive surgical staging for uterine cancers can be adequately performed through a PI without greater morbidity or mortality. By using this surgical approach, patients with uterine cancer can benefit from the inherent benefits previously described for PI. Appropriate patient selection, however, is necessary. PMID- 12648595 TI - Altered imprinting, promoter usage, and expression of insulin-like growth factor II gene in gestational trophoblastic diseases. AB - OBJECTIVE: We aimed to understand the involvement of imprinted genes in the pathogenesis of gestational trophoblastic diseases (GTD) such as hydatidiform mole (H-mole) and gestational trophoblastic tumors (GTT). METHODS: An allelic typing assay was performed using a PCR-RFLP-based method for identification of heterozygous informative cases. The usage of insulin-like growth factor-II (IGF2) promoters was examined by RT-PCR using promoter-specific primers. The mRNA expression of IGF2 and H19 was quantified using a densitometer. RESULTS: The imprinting of IGF2 and H19 was maintained in all normal placenta tissues (n = 15) but relaxed in GTD (n = 47). Loss of imprinting (LOI) of IGF2 was in the order of GTT (57%) > complete H-mole (43%) > partial H-mole (25%). Similarly, LOI of H19 was in the order of GTT (40%) > complete H-mole (18%) > partial H-mole (0%). Promoter usage pattern of IGF2 changed with gestation stage of normal placentae and GTD. In normal placentae, the usage of promoter P1 was higher than that of P4 in the first trimester but lowered in the full term. H-mole and GTT predominantly used promoter P1 with relative silencing of promoter P4. Although normal early placenta and various GTD tissues showed the similar usage of IGF2 promoter P1, GTT tissues revealed the higher expression levels of IGF2 but a down-regulation of H19 relative to the normal early placentae. CONCLUSIONS: These results suggest that LOI, deregulation of IGF2 promoters, and the altered expression levels of IGF2 and H19 genes might be associated with the progression of GTD. PMID- 12648596 TI - Radical vaginal trachelectomy and pelvic lymphadenectomy for preservation of fertility in early cervical carcinoma. AB - OBJECTIVE: The aim of this study was to examine our experience with radical vaginal trachelectomy in women with early cervical cancers who desire to maintain fertility. METHODS: Women who underwent radical vaginal trachelectomy with pelvic lymphadenectomy over a 6-year period are the basis of this report. Subjects were selected for this treatment on the basis of favorable cervical tumors and a desire to maintain fertility. All subjects were informed that this therapy did not represent standard treatment for early stage cervical cancer. Obstetrical and oncologic outcomes were evaluated. RESULTS: Twenty-one women underwent this procedure. The median age was 30 years (range 23-41); 14 were nulligravid and 16 were nulliparous. Mean tumor diameter was 1.1 cm (range 0.3-3.0). Mean operative time was 318 min, with a mean blood loss of 293 cc, and average hospital stay was 3 days. Three patients had transient neuropathy postoperatively. No patient required laparotomy. Two patients had completion of radical vaginal hysterectomy for an inability to clear the cancer with trachelectomy and 1 had postoperative radiation for high-risk features on final pathology. With an average follow-up of 31.5 months, there have been no recurrences. Three women have become pregnant: 1 woman delivered twins at 24 weeks, 1 woman delivered a singleton at term, and 1 patient had rupture of membranes and chorioamnionitis at 20 weeks gestation. CONCLUSIONS: Radical vaginal trachelectomy with pelvic lymphadenectomy permits preservation of fertility in selected patients. To date, with more than 150 cases reported in the literature, recurrence rates are comparable to those seen with radical hysterectomy. PMID- 12648597 TI - Protein profiling of complete mole and normal placenta using ProteinChip analysis on laser capture microdissected cells. AB - INTRODUCTION: Surface-enhanced laser desorption/ionization mass spectrometry (SELDI-MS) is a novel method for biomarker discovery that can provide a rapid protein expression profile from a variety of biological samples. Since SELDI-MS requires a small amount of biological material, this technique is ideal for analyzing proteins isolated from microdissected tissue samples. The current study was undertaken to investigate potential differences in protein expression between normal and molar trophoblast procured by laser capture microdissection (LCM) utilizing SELDI ProteinChip array technology. Further knowledge of protein expression in complete mole may advance our understanding of the pathogenesis of gestational trophoblastic diseases. MATERIALS AND METHODS: Laser capture microdissected trophoblast cells from nine fresh complete moles were analyzed and compared to the trophoblast cells from 10 fresh normal placentas of comparable gestational age, using SELDI ProteinChip to identify potential differences in protein expression. RESULTS: Three metal binding polypeptides were identified with the estimated molecular weights of 11.3, 13.8, and 14.0 kDa, which appeared in significantly lower levels in complete mole as compared to normal trophoblast cells (P < 0.001, P < 0.03, and P < 0.01). DISCUSSION: While further characterization of these protein peaks is important and necessary, our current work clearly demonstrates that the combined technology of SELDI and LCM is effective in distinguishing protein expression between normal placenta and complete mole. Further knowledge of protein expression in complete mole may advance our understanding of molecular mechanisms and improve management in gestational trophoblastic diseases. PMID- 12648598 TI - Inhibition of paclitaxel-induced apoptosis by the specific COX-2 inhibitor, NS398, in epithelial ovarian cancer cells. AB - OBJECTIVE: In vitro studies have revealed that treatment of various human cancer cell lines with specific cyclo-oxygenase 2 (COX-2) inhibitors induces apoptotic cell death. It is currently proposed that the combination of COX-2 inhibitors with chemotherapeutic agents improves the efficacy of cancer treatment. MATERIALS AND METHODS: In this study we sought to determine the effects of combining paclitaxel and the COX-2 inhibitor NS398 on apoptosis of epithelial ovarian cancer (EOC) cells. Two EOC cell lines, SKOV3 and MDAH2774, were exposed to increasing concentrations of paclitaxel (0.1, 10, and 100 microM) and NS398 (10, 100 microM) as well as a combination of both drugs. Apoptosis was evaluated by the Tunel assay. The fluorescein-labeled DNA was visualized directly by fluorescence microscopy and quantitated by flow cytometry. RESULTS: While NS398 did not significantly alter apoptosis of either EOC cell lines after 24 h of continuous exposure, treatment of both cell lines with paclitaxel resulted in a significant increase in the rate of apoptosis (60-70%). Concomitant treatment of both SKOV3 and MDAH2774 cells with paclitaxel and NS398 resulted in marked impairment of paclitaxel-induced apoptosis. Similarly, sequential treatment during which both cell lines were treated with NS398 for 4 h, triple-washed, and then exposed to paclitaxel for 24 h resulted in a significant inhibition of paclitaxel-induced apoptosis. Similar inhibition was seen when NS398 was replaced by aspirin. CONCLUSIONS: Combining COX-2 inhibitors and paclitaxel does not have an additive or synergistic tumoricidal effect. On the contrary, NS398 treatment markedly inhibited the apoptotic effects of paclitaxel in each of these two EOC cell lines. PMID- 12648599 TI - The use of antioxidant therapies during chemotherapy. AB - OBJECTIVE: At the present time, many cancer patients combine some form of complementary and alternative medicine therapies with their conventional therapies. The most common choice of these therapies is the use of antioxidants. RESULTS: A review of four common antioxidants is undertaken, which includes vitamin E (mixed tocopherols and tocotrienols), beta-carotene (natural mixed carotenoids), vitamin C (ascorbic acid), and vitamin A (retinoic acid). Antioxidants act as electron acceptors as well as therapeutic biologic response modifiers. Despite the fact that chemotherapy-induced formation of free radicals is well-demonstrated, chemotherapy-induced cytotoxicity in general does not seem to depend on formation of reactive oxygen species. CONCLUSIONS: Currently, evidence is growing that antioxidants may provide some benefit when combined with certain types of chemotherapy. Because of the potential for positive benefits, a randomized controlled trial evaluating the safety and efficacy of adding antioxidants to chemotherapy in newly diagnosed ovarian cancer is underway at the University of Kansas Medical Center. PMID- 12648600 TI - Clinically inapparent invasive vulvar carcinoma in an area of persistent Paget's disease: a case report. AB - BACKGROUND: Invasive vulvar carcinoma is reported to occur in 5 to 20% of patients with vulvar Paget's disease. We report a case in which a clinically inapparent invasive lesion was discovered on reexcision of microscopically persistent vulvar Paget's disease. CASE: A 58-year-old woman presented with a diagnosis of vulvar Paget's disease. A wide local excision of the lesion was performed and pathologic analysis revealed microscopic Paget's disease at two of the margins. The patient returned for a follow-up 4 months later and a vulvar biopsy revealing persistent Paget's cells was obtained from the area of the prior microscopically positive surgical margin. A reexcision was performed from the normal-appearing vulva and invasive vulvar carcinoma was noted in this specimen. CONCLUSIONS: This case demonstrates several concerning aspects of this disease, most important of which is that the clinically apparent lesion did not contain the clinically significant invasive lesion. Invasive vulvar carcinoma may occur in association with microscopically persistent vulvar Paget's disease, a condition often encountered after primary treatment with wide local excision. PMID- 12648601 TI - Primary in situ squamous cell carcinoma of the endometrium, with extensive squamous metaplasia and dysplasia. AB - BACKGROUND: Primary squamous cell carcinoma of the endometrium is exceedingly rare. It has been described in association with pyometra but its etiology is still unclear. CASE: The authors report the case of a 75-year-old woman who presented with pelvic pain and pyometra. No gross tumor was identified in the uterus; however, extensive epidermalization of the endometrial mucosa was noted. Microscopic findings were consistent with a primary in situ squamous carcinoma of the endometrium associated with extensive squamous metaplasia and areas of dysplasia. HPV antigen and DNA detection were negative in both the endometrial lesions and the cervix. CONCLUSIONS: These results support the sequence of change with squamous metaplasia, progressing through dysplasia to carcinoma as a possible pathogenetic process. HPV's role, however, remains uncertain. PMID- 12648602 TI - Successful management of endodermal sinus tumor of the ovary associated with pregnancy. AB - BACKGROUND: Pregnancy complicated by endodermal sinus tumor of the ovary has rarely been reported. CASE: A 32-year-old pregnant woman was found to have an ovarian tumor. At 19 weeks of gestation, tumorectomy was performed and a diagnosis of primary endodermal sinus tumor of the ovary (stage Ic) was made. Pregnancy was continued without postoperative chemotherapy. At 36 weeks of gestation, she underwent cesarean section combined with second-look laparotomy. A normal infant was delivered and there were no signs of recurrence. Subsequently, three courses of combination chemotherapy with bleomycin, etoposide, and cisplatin were administered. There was no evidence of recurrence at 27 months after initial treatment. CONCLUSIONS: Successful management of endodermal sinus tumor of the ovary in a pregnant woman is reported. PMID- 12648603 TI - Primary small cell carcinoma of the vagina. AB - BACKGROUND: Primary vaginal small cell carcinoma is extremely rare, with a total number reported in English-language journals to date of 23. Most patients die of the disease within 2 years of diagnosis from metastatic disease. CASE: A 69-year old woman presented with vaginal spotting while on Premarin. She was subsequently diagnosed with Stage I (T1N0M0) small cell carcinoma of the vagina. She underwent concurrent chemoradiation and then brachytherapy for persistent disease. Due to residual disease after the brachytherapy, surgical resection was planned but aborted because of metastatic disease. CONCLUSIONS: Of the three reported cases treated with concurrent chemoradiation, ours is the first case reported with persistent local disease after therapy. Extrapolating from the available clinical trials from lung carcinoma, concurrent chemoradiation as a primary treatment approach should still be considered. PMID- 12648604 TI - A case of Mullerian adenosarcoma of the uterus treated with liposomal doxorubicin. AB - OBJECTIVE: We report on a case of uterine a denosarcoma responsive to treatment with liposomal doxorubicin (Doxil). METHODS: The clinical course, histopathology, and radiologic studies of the case were reviewed and are reported. RESULTS: A 69 year-old woman presented to our institution with dehydration and failure to thrive, and recurrent uterine adenosarcoma, 9 months after initial diagnosis. She had undergone total abdominal hysterectomy and bilateral salpingo-oophorectomy at the time of diagnosis, followed by two cycles of Ifosfamide and treatment with megestrol acetate. The patient presented to our institution for hospice care placement, severely dehydrated and with lethargy and with recurrent disease in the pelvis and vagina. She underwent treatment with six cycles of liposomal doxorubicin with a marked response. Given the development of Grade II hand-foot syndrome, liposomal doxorubicin was stopped and two cycles of carboplatin/paclitaxel chemotherapy were administered. Given the patient's disease progression on this regimen, surgical cytoreduction for localized recurrent disease was then performed. The patient developed disease recurrence after a 7-month disease-free interval and 2 years after initial diagnosis. Following another two cycles of liposomal doxorubicin, the patient underwent another cytoreductive procedure for recurrent disease. Two months later, the patient expired, 29 months after diagnosis and 20 months after initial treatment with liposomal doxorubicin chemotherapy. CONCLUSIONS: Liposomal doxorubicin appears to be active in the treatment of recurrent uterine adenosarcoma. PMID- 12648605 TI - Endometrial stromal sarcoma with a sole t(X;17) chromosome change: report of a case and review of the literature. AB - BACKGROUND: Endometrial stromal sarcomas (ESSs) exhibit varying degrees of malignancy and heterogeneity at the karyotypic level. The biological mechanisms that contribute to tumorigenesis of ESS are still largely unknown. CASE: A 33 year-old woman suffering from ESS was treated primarily surgically. Cytogenetic evaluation of the primary uterine nodule and metastatic tumor showed 46,XX,t(X;17)(p11:q23) karyotype in all metaphases analyzed. Normal endometrial cells exhibited 46,XX karyotype. Fluorescence in situ hybridization analysis confirmed the presence of the reciprocal t(X;17) translocation and allowed for the positioning of the chromosome X breakpoint distal to SSX1 gene loci. CONCLUSIONS: Our report of a previously undescribed sole cytogenetic translocation in an advanced stage of ESS might identify a cytogenetically distinct subgroup of ESS and help to reveal genes involved in ESS tumorigenesis. PMID- 12648606 TI - Immunohistochemical determination of hormone receptors in uterine adenosarcomas. PMID- 12648608 TI - Characterization of properties underlying rhythmicity in mouse portal vein. AB - We have used sharp intracellular and patch clamp electrophysiology, together with mechanical recordings and immunohistochemistry to characterize some of the properties underlying spontaneous rhythmicity in isolated murine portal vein. Mechanical recordings revealed that isolated whole portal veins were spontaneously active and generated regular contractions every 5-15-s that persisted in the presence of cyclopiazonic acid (CPA) (10 microM) or thapsigargin (100 nM). Intracellular recordings from smooth muscle cells revealed spontaneous depolarizations (SDs) in membrane potential, which were abolished by nifedipine (1 microM). Whole cell patch clamp recordings from isolated smooth muscle cells revealed an inward "pacemaker" current (I(H)) at negative potentials. Immunohistochemical studies failed to detect the presence of Kit-immunoreactive cells in portal veins of wild type mice, but were consistently observed in the small intestine. Furthermore, portal veins obtained from W/W(v) mutant mice, which lack full expression of the tyrosine-kinase, c-Kit, were also rhythmically active and were not different from wild type mice, in either their electrical or mechanical properties. These results show that both the wild type and W/W(v) mutant mouse portal vein are rhythmically active in vitro. However, pacemaker activity in this blood vessel occurs in the absence of Kit-immunoreactive cells; and is not critically dependent upon release of Ca(2+) from intracellular stores. The rhythmic pacemaker activity of mouse portal vein does involve L-type Ca(2+) currents, and possibly pacemaker conductances intrinsic to the smooth muscle. PMID- 12648609 TI - The influence of vasopressin on tonic activity of cardiovascular neurones in the ventrolateral medulla of the hypertensive rat. AB - In normotensive Wistar rats, vasopressin may act as an excitatory neurotransmitter at synapses of paraventricular neurones on rostral ventrolateral medullary vasomotor neurones. We studied the influence of this neurotransmitter in spontaneously hypertensive rats to determine if it contributed to the increases in sympathetic nerve traffic in these rats.A five-barrel micropipette assembly was used for extracellular recording of neuronal activity and for microiontophoresis of drugs into the vicinity of identified medullary vasomotor neurones. Excitatory effects of iontophoretically applied vasopressin were blocked by simultaneous iontophoretic application of V(1a) antagonist. Similar application of the vasopressin receptor antagonist did not block an excitatory effect of iontophoretically applied glutamate. Excitatory effects produced by activating paraventricular neurones were also blocked by the V(1a) antagonist. However, the vasopressin antagonist did not alter the ongoing activity of medullary vasomotor neurones. Therefore, in these anaesthetised hypertensive rats, we concluded that vasopressin neurones do not exert a significant tonic drive to rostral ventrolateral medullary-spinal vasomotor neurones. PMID- 12648611 TI - Cellular localization and distribution of neurokinin-1 receptors in the rat stomach. AB - In the stomach, the majority of substance P's effects are mediated by the activation of neurokinin-1 (NK1) receptors. The gastric cellular distribution of these receptors in Wistar and Sprague-Dawley rats was determined using immunocytochemistry. The localization of the NK1 receptors with respect to von Willebrand's factor, protein gene product 9.5, substance P, vasoactive intestinal peptide, and calcitonin gene-related peptide was also determined. Results show that NK1 receptor immunoreactivity was dependent on the duration of fixation. In corpus and antrum tissues that were fixed in 4% paraformaldehyde for 30 min, the presence of NK1 receptor immunoreactivity was demonstrated on nerve fibers throughout the stomach, on the surface and in the cytoplasm of myenteric cell bodies, on circular smooth muscle cells, and on vascular endothelial cells. This was observed in tissues from both rodent strains. Overnight fixation in the same fixative, however, demonstrated the presence of NK1 receptor immunoreactivity only on nerve fibers and cell bodies of the myenteric plexus, and on circular smooth muscle cells. In 30-min fixed tissues, the localization of NK1R immunoreactivity on vascular endothelial cells and nerve fibers was confirmed by co-localization with von Willebrand's factor and protein gene product 9.5 immunoreactivity, respectively. In both rodent strains, NK1 receptor immunoreactivity was co-localized with substance P immunoreactivity on nerve fibers of the longitudinal and circular muscle. In the Wistar rat, NK1 receptor immunoreactivity was co-localized with vasoactive intestinal peptide immunoreactivity or calcitonin gene-related peptide immunoreactivity throughout the stomach. However, in the Sprague-Dawley rat, NK1 receptor immunoreactivity was only co-localized with calcitonin gene-related peptide immunoreactivity in a minority of fibers of the circular muscle. The overall results of this study show that the antigenic epitopes of the NK1 receptor are sensitive to overfixation. When tissues were not overfixed, NK1 receptor immunoreactivity was distributed more extensively throughout the rat stomach than has been described previously. The results of this study provide the anatomical basis for many of the actions of substance P in the rat stomach. PMID- 12648610 TI - Impact of renal denervation on renal content of GLUT1, albuminuria and urinary TGF-beta1 in streptozotocin-induced diabetic rats. AB - In long-term diabetes mellitus, the progression of nephropathy has been related to the occurrence of autonomic neuropathy. This study was designed to evaluate the effects of bilateral denervation of the kidneys of streptozotocin-diabetic rats, an experimental model that presents diabetic nephropathy with increased abundance of cortical GLUT1 in the kidney and increased urinary excretion of albumin and transforming growth factor-beta1 (TGF-beta1). Twenty-four-hour urinary TGF-beta1 (ELISA), urinary albumin (electroimmunoassay) and GLUT1 protein levels (Western blotting) in the renal cortex and medulla were evaluated in diabetic (n=13) and control (n=13) rats 45 days after streptozotocin injection, submitted or not to surgical renal denervation. Evaluations were performed 15 days after the surgery. The effects of renal denervation were confirmed by intra renal decrease of norepinephrine levels. Mean arterial pressure did not differ between diabetic and control rats, whether they underwent renal denervation or not. Renal denervation increased cortical (6905+/-287, 3506+/-193, 4144+/-246 and 5204+/-516 AU in renal-denervated controls, controls, renal-denervated diabetics and diabetics, respectively) and medullar GLUT1 protein in control rats, but reverted the cortical GLUT1 protein rise determined by diabetes. Although kidney denervation in diabetic rats induced a decrease in cortical GLUT1 abundance toward normal levels, these levels did not reach those of normal animals. However, renal denervation did not determine any changes in urinary albumin and urinary TGF-beta1 in both diabetic (127.3+/-12 microg/24 h and 111.8+/-24 ng mg( 1) creatinine, respectively) and control rats (45.9+/-3 microg/24 h and 13.4+/-4 ng mg(-1) creatinine, respectively). In conclusion, early-phase renal denervation in streptozotocin-diabetic rats produces a normalisation of previously elevated cortical GLUT1 protein content, but is not enough for reverting the increased urinary TGF-beta1 and albuminuria of diabetes. PMID- 12648613 TI - Local cooling alters neural mechanisms producing changes in peripheral blood flow by spinal cord stimulation. AB - This study was performed to investigate the respective role of sensory afferent and sympathetic fibers in peripheral vasodilatation induced by spinal cord stimulation at different hindpaw skin temperatures. Cooling the skin was used as a strategy to enhance sympathetic activity [Am. J. Physiol.: Heart Circ. Physiol. 263 (1992) H1197]. Cutaneous blood flow in the footpad of anesthetized rats was recorded using laser Doppler flowmetry. Local cooling (<25 degrees C) or moderate local cooling (25-28 degrees C) of the hindpaw was produced with a cooling copper coil. Spinal cord stimulation delivered at clinically relevant parameters and with 30%, 60%, and 90% of motor threshold induced the early phase of vasodilatation in the cooled and the moderately cooled hindpaw. In addition, spinal cord stimulation at 90% of motor threshold produced the late phase of vasodilatation only in the cooled hindpaw, which was possible to block by the autonomic ganglion-blocking agent, hexamethonium. The early responses to spinal cord stimulation in the moderately cooled hindpaw were not affected by hexamethonium. In contrast, both the early and the late phase responses were eliminated by CGRP (8-37), an antagonist of the calcitonin gene-related peptide receptor. After dorsal rhizotomy, spinal cord stimulation at 90% of motor threshold elicited hexamethonium-sensitive vasodilatation in the cooled hindpaw (late phase). These results suggest that spinal cord stimulation-induced vasodilatation in the cooled hindpaw (<25 degrees C) is mediated via both the sensory afferent (early phase of vasodilatation) and via suppression of the sympathetic efferent activity (late phase) although the threshold for vasodilatation via the sympathetic efferent fibers is higher than that via sensory nerves. In contrast, vasodilatation via sensory afferent fibers may predominate with moderate temperatures (25-28 degrees C). Thus, two complementary mechanisms for spinal cord stimulation-induced vasodilatation may exist depending on the basal sympathetic tone. PMID- 12648612 TI - Control of canine membranous urethra, bulbocavernosus and ischiocavernosus muscles by lumbosacral sympathetic pathways. AB - The membranous urethral muscles, bulbocavernosus muscle (BCM) and ischiocavernosus muscle (ICM) play a significant role on expulsion at ejaculation. Projection of the sympathetic pathways in the retroperitoneum to these muscles was investigated in the dog. Intraluminar pressure of the membranous urethra and contractions of the BCM and ICM to electrical stimulation of the lumbar splanchnic nerve (LSN) or the lumbosacral sympathetic chain (LSC) were examined. Stimulation of the LSN was performed after transection of the unilateral hypogastric nerve (HGN) to clarify its cross-innervation. Stimulation of each of the 2nd to 4th LSNs elicited elevation of the intraluminal pressure of the membranous urethra. In eight dogs whose right HGNs were transected, 15 of the 19 right LSNs, and 12 of the 16 left LSNs examined elicited elevation of the membranous urethral pressure, respectively. In five dogs whose left HGNs were transected, 6 of the 9 right LSNs and 10 of the 12 left LSNs stimulated elicited elevation of the pressure. Stimulation of the HGN caused no detectable contractions of the BCM and ICM in all six dogs examined. Stimulation of the LSC elicited responses of the BCM and ICM in all six dogs examined and the membranous urethra in three of six dogs. The maximum value of the response of the BCM was greater than that of the ICM. After consecutive transection of the ipsilateral pelvic nerve, stimulation of the LSC showed no significant changes in the responses of the BCM and ICM. The above results indicate the following: (1). each of the L2-L4 LSNs sends signals to the membranous urethra via two routes, passing through the ipsilateral or contralateral HGN; (2). the above nerve does not send effective signals to the BCM and ICM; (3). the LSC sends signals to the membranous urethra, BCM and ICM; (4). the signals from the LSC mainly pass through the pudendal nerve to the BCM and ICM; and (5). the signals were dominant to the BCM compared with the ICM. PMID- 12648614 TI - Responses and afferent pathways of C(1)-C(2) spinal neurons to gastric distension in rats. AB - Some evidence shows that the upper cervical spinal cord might play an important role in propriospinal processing as a sensory filter and modulator for visceral afferents. The aims of this study were to determine (1). the responses of C(1) C(2) spinal neurons to gastric distension and (2). the relative contribution of vagal and spinal visceral afferent pathways for transmission of gastric input to the upper cervical spinal cord. Extracellular potentials of single C(1)-C(2) spinal neurons were recorded in pentobarbital anesthetized male rats. Graded gastric distension (20-80 mm Hg) was produced by air inflation of a latex balloon surgically placed in the stomach. Sixteen percent of the neurons (32/198) responded to gastric distension; 17 neurons were excited and 15 neurons were inhibited by gastric distension. Spontaneous activity of neurons with inhibitory responses was higher than those neurons with excitatory responses (18.1+/-2.7 vs. 3.8+/-1.7 impulses s(-1), p<0.001). Twenty-eight of thirty-two (87.5%) neurons responded to mechanical stimulation of somatic fields on head, neck, ears or shoulder. Most lesion sites of neurons with excitatory responses were found in laminae V, VII; however, neurons with inhibitory responses were in laminae III, IV. Bilateral cervical vagotomy abolished responses of 4/8 neurons tested. Spinal transection at C(6)-C(7) abolished responses of the other four neurons that still responded to gastric distension after bilateral vagotomy. Results of these data supported the concept that a group of C(1)-C(2) spinal neurons might play a role in processing sensory information from the stomach that travels in vagal and spinal visceral afferent fibers. PMID- 12648615 TI - Arterial sympathetic innervation and cerebrovascular diseases in original rat models. AB - The role of the arterial sympathetic innervation in cerebrovascular pathology was investigated in new experimental models using Brown Norway (BN) and Long-Evans (LE) rats. The BN rat is susceptible to intracerebral hemorrhage (ICH) within the cerebral cortex when rendered hypertensive whereas the LE rat is prone to cerebral aneurysms (CAs) in arteries of the circle of Willis with hypertension and carotid ligation. Noradrenaline (NA) content, determined by high performance liquid chromatography (HPLC), was lower both in the caudal and cerebral arteries in the BN than in the LE rat. Denervation of cerebral arteries by superior cervical ganglionectomy did not increase ICH lesion incidence in BN hypertensive rats. A possible link between the level of caudal artery NA content and the occurrence of ICH lesions and CAs was studied in rats from two distinct BNXLE crosses: back-cross (BC) rats (F1XBN) and F2 rats (F1XF1) which respectively display, with hypertension and carotid ligation, a high incidence of either ICH lesions or CAs. In BC rats, the level of caudal artery NA content was not related to ICH lesion occurrence. However, in F2 rats a low caudal artery NA content was associated with a high incidence of ruptured CAs. Thus, a low arterial sympathetic innervation may participate in mechanisms leading to rupture of CAs. PMID- 12648617 TI - Cell destruction from intraocular use of benzalkonium chloride. PMID- 12648618 TI - Incision burns. PMID- 12648620 TI - Transparent irrigation cannula. PMID- 12648622 TI - Why mix trypan blue with viscoelastic agents? PMID- 12648623 TI - Consultation section. Cataract surgical problem. PMID- 12648631 TI - Phacoemulsification in eyes with corneal opacification. AB - We describe a technique in which transcorneal illumination with a fiber-optic light probe is used to safely perform phacoemulsification in cataractous eyes with dense corneal opacification. The technique was used in 3 eyes of 3 patients. No surgical complications occurred, and all eyes had improved visual acuity. Indications and further refinement of the technique and instrumentation are discussed. PMID- 12648632 TI - Single-action implantation of a 3-piece acrylic intraocular lens with an injector. AB - We report a method to insert a 3-piece AcrySof MA30BA acrylic foldable intraocular lens (IOL) (Alcon) in a single action using a Monarch II (Alcon) injector. The technique was used in 134 eyes. The incision widths ranged from 3.00 to 3.75 mm. The IOLs were successfully inserted in the capsular bag. Complications included cracks in the IOL optic in 3 eyes (3%), haptic damage in 3 eyes (3%), and inadequate self-sealing of the incision in 18 eyes (13%). PMID- 12648633 TI - Histopathology of corneal melting associated with diclofenac use after refractive surgery. AB - PURPOSE: To describe the histopathology of the cornea in 3 cases of corneal melting associated with diclofenac therapy after refractive surgery procedures. SETTING: Clinic and pathology laboratory. METHODS: Three cases of corneal melting associated with diclofenac therapy (2 after laser in situ keratomileusis [LASIK] and 1 after mini-radial keratectomy enhancement of a LASIK undercorrection) were studied using patient and referring physician interviews, chart reviews, and histopathologic examination of the corneal tissue. RESULTS: In all 3 cases, the flaps were dislocated and the stromal corneal bed was exposed. Diclofenac, generic or brand name, was used in all cases; in 1 case, both generic and brand name were used. Dosing and duration varied, but in all 3 cases diclofenac was used at least 4 times a day for at least 3 days after LASIK. Topical steroids were also prescribed, but 1 patient did not use them. Preoperative medical conditions were present in 2 cases. Histologic analysis showed evidence of an inflammatory response in advanced cases and keratolysis and lack of inflammatory cells in the flaps that were amputated early. CONCLUSIONS: The use of generic or brand-name diclofenac with or without adjunctive topical steroids after LASIK can be associated with corneal melting when the LASIK flap is dislodged and the corneal stromal bed exposed. Caution is recommended with diclofenac use after LASIK in such cases. PMID- 12648634 TI - Maximum permissible lateral decentration in aberration-sensing and wavefront guided corneal ablation. AB - PURPOSE: To investigate the lateral alignment accuracy needed in wavefront-guided refractive surgery to improve the ocular optics to a desired level in a percentage of normally aberrated eyes. SETTING: Department of Ophthalmology, University of Zurich, Zurich, Switzerland. METHODS: The effect of laterally misaligned ablations on the optical outcome was simulated using measured wavefront aberration patterns from 130 normal eyes. The calculations were done for 3.0 mm, 5.0 mm, and 7.0 mm pupils. The optical quality of the simulated correction was rated by means of the root-mean-square residual wavefront error. RESULTS: To achieve the diffraction limit in 95% of the normal eyes with a 7.0 mm pupil, a lateral alignment accuracy of 0.07 mm or better was required. An accuracy of 0.2 mm was sufficient to reach the same goal with a 3.0 mm pupil. CONCLUSION: Procedures must be developed to ensure that the ablation is within a tolerance range based on each eye's original optical error. Rough centration based on the surgeon's judgment might not be accurate enough to achieve significantly improved optical quality in a high percentage of treated eyes. PMID- 12648635 TI - Laser in situ keratomileusis flap suturing to treat recalcitrant flap striae. AB - PURPOSE: To describe the management of resistant laser in situ keratomileusis (LASIK) flap striae using interrupted sutures. SETTING: Cullen Eye Institute, Baylor College of Medicine, Houston, Texas, USA. METHODS: Interrupted sutures were used to treat striae in the LASIK flaps in 7 eyes of 7 patients. The mean interval from LASIK to the time of suturing was 11.8 months (range 3.0 to 29.0 months). The preoperative best spectacle-corrected visual acuity (BSCVA) was 20/20(-2) to 20/100. All patients had subjective visual complaints with spectacle correction, and all corneas had obvious macrostriae on slitlamp biomicroscopic examination. RESULTS: The mean patient follow-up was 7.0 months (range 1.5 to 14.0 months). Six patients regained a BSCVA of 20/20 or better with resolution of preoperative irregular astigmatism and subjective visual disturbance. In 1 patient, mild central striae recurred. CONCLUSION: Flap suturing was an effective means of treating visually significant flap striae. PMID- 12648636 TI - Choroidal neovascularization in phakic eyes with anterior chamber intraocular lenses to correct high myopia. AB - PURPOSE: To analyze the appearance, incidence, and characteristics of choroidal neovascularization (CNV) in patients with high myopia corrected by implantation of a phakic anterior chamber intraocular lens (PACL). SETTING: University Miguel Hernandez, Instituto Oftalmologico de Alicante, Alicante, Spain. METHODS: The CNV observed in 294 consecutive eyes (181 patients) implanted with a PACL for the correction of high myopia (-7.0 to -38.0 diopters) was studied. The mean follow up was 50.6 months +/- 32.8 (SD) (range 6 to 120 months). RESULTS: Choroidal neovascularization occurred in 5 eyes (1.70%); 3 eyes were in women, and 2 were in men. The interval between PACL implantation and CNV was 63.2 +/- 27.3 months (range 18 to 87 months). The CNV was subfoveal in 4 eyes and juxtafoveal in 1 case. The mean best spectacle-corrected visual acuity (BSCVA) after PACL implantation and before the appearance of CNV was 0.53 +/- 0.18 (range 0.4 [20/50] to 0.8 [20/25]); after CNV appeared, it was 0.26 +/- 0.18 (range 0.05 [20/400] to 0.5 [20/40]), a statistically significant difference (P =.001, paired Student t test). In 2 cases, the CNV was treated with photodynamic therapy (PDT); in the other 3 cases, PDT was rejected. The cumulative risk for CNV (Kaplan-Meier survival analysis) in highly myopic patients corrected by PACL implantation was 0.43% at 18 months and 5.4% at 87 months. CONCLUSIONS: Implantation of a PACL to correct high myopia was followed by a small incidence of CNV (cumulative risk of 5.4% at 87 months). The appearance of CNV was followed by a significant decrease in BSCVA. PMID- 12648637 TI - Effect of pupil size on visual function under monocular and binocular conditions in LASIK and non-LASIK patients. AB - PURPOSE: To compare binocular and monocular vision in patients treated with laser in situ keratomileusis (LASIK) and in non-LASIK patients. SETTING: Jules Stein Eye Institute, Los Angeles, California, USA. METHODS: This comparative cross sectional study comprised 20 postoperative LASIK patients and 20 non-LASIK ametropic patients. LogMAR visual acuity, contrast sensitivity, and infrared pupillometry were tested. Outcome measures were better-eye monocular acuity, binocular acuity, better-eye contrast sensitivity, binocular contrast sensitivity, and pupil diameter under monocular and binocular conditions. RESULTS: Binocular visual acuity and contrast sensitivity were statistically significantly better than the visual acuity in the better eye (P =.0047 to <.0001) in both patient groups. Pupil diameter was statistically significantly smaller under the binocular condition than the monocular condition (P <.0001) in both groups. CONCLUSIONS: Monocular testing induced larger pupil diameters, which was associated with reduced vision compared to binocular measurements in LASIK and non-LASIK patients. PMID- 12648638 TI - Amniotic membrane transplantation for bullous keratopathy in eyes with poor visual potential. AB - PURPOSE: To evaluate the long-term outcomes of epithelial debridement and amniotic membrane transplantation (AMT) for pain and discomfort relief in patients with symptomatic bullous keratopathy and poor visual potential. SETTING: Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami, Miami, Florida, USA. METHODS: This retrospective study included 18 eyes (18 patients) with bullous keratopathy presenting with intractable pain or discomfort and poor visual potential. After epithelial debridement, all eyes had AMT with the basement membrane side up. During a mean follow-up of 25.1 months +/- 9.6 (SD) (range 12 to 45 months), pain relief, epithelial healing, and visual changes were analyzed. RESULTS: Pain relief was obtained in 88% of patients. Sixty-six percent of eyes had complete resolution of ocular discomfort starting soon after the first postoperative day. One eye had evisceration for persistent pain 10 months postoperatively. Corneal epithelial healing was complete in all except 1 eye. Remaining complaints included foreign-body sensation (5%), tearing (11%), and photophobia (5%). CONCLUSIONS: Amniotic membrane transplantation was a safe, effective, and long-lasting treatment modality for intractable pain associated with chronic bullous keratopathy in eyes with poor visual potential. It can be an alternative to conjunctival flaps for the long-term management of patients with bullous keratopathy in whom corneal transplantation is not indicated. A comparison of the efficacy of AMT with that of other surgical procedures must be performed. PMID- 12648640 TI - Four-point fixation of posterior chamber intraocular lenses in children with unilateral aphakia. AB - PURPOSE: To evaluate the results of 4-point scleral fixation of posterior chamber intraocular lenses (PC IOLs) in children. SETTING: Mansoura Ophthalmic Center, Mansoura University, Mansoura, Egypt. METHODS: This retrospective study comprised 20 aphakic eyes of 20 children. Preoperative investigations proved the absence of adequate posterior capsule support. All patients had ab externo 4-point scleral fixation of a PC IOL. All preoperative and postoperative data were studied and analyzed. RESULTS: The mean preoperative age was 7.7 years (range 4 to 11 years). The mean follow-up was 19.35 months (range 13 to 30 months). The preoperative findings included a best corrected visual acuity of 6/18 or better in 8 eyes (40%), corneal scarring in 18 eyes, sector iridectomy in 7 eyes, and after cataract in 4 eyes. The intraoperative complications included vitreous hemorrhage in 2 eyes and slippage of 1 suture in 1 eye. Postoperative complications included mild anterior uveitis in 12 eyes and severe fibrinoid reaction in the anterior chamber in 8 eyes. There were no cases of suture exposure, IOL malpositioning, or infection. Twelve patients (60%) had a visual acuity of 6/18 or better. The cause of the poor visual outcome in many patients was preexisting amblyopia. CONCLUSIONS: The results suggest that 4-point scleral fixation is an option to correct aphakia in children. However, long-term follow-up is important to assess the procedure's safety. PMID- 12648639 TI - Morphological and functional results of AcrySof intraocular lens implantation in children: prospective randomized study of age-related surgical management. AB - PURPOSE: To evaluate the prevalence and severity of posterior capsule opacification (PCO) in pediatric eyes with a foldable acrylic AcrySof (Alcon) intraocular lens (IOL) and age-related surgical methods. SETTING: Department of Ophthalmology, University of Vienna, Medical School, Vienna, Austria. METHODS: This prospective randomized study comprised 50 eyes of 34 children aged between 2 and 16 years. Eyes of children between 2 and 5.9 years were consecutively randomized to Group 1a (primary posterior capsulotomy and anterior vitrectomy) or Group 1b (optic capture in addition). Eyes of children between 6 and 16 years were consecutively randomized to Group 2a (primary posterior capsulotomy without anterior vitrectomy), Group 2b (optic capture in addition), or Group 2c (in-the bag IOL implantation without opening the posterior capsule). Main outcome parameters were the incidence and severity of PCO formation, early postoperative complications, pigmented cell deposits on the IOL surface, and cataract morphology. RESULTS: The visual axis was clear at the last follow-up in all eyes in Groups 1a, 1b, 2a, and 2b except in 1 eye in Group 1a. Sixty-percent of eyes in Group 2c had PCO. The incidence of early postoperative complications was significantly higher in eyes that developed PCO than in those that maintained a clear visual axis. There was no evidence that cataract morphology influenced PCO rates. CONCLUSIONS: The AcrySof IOL was well tolerated in pediatric eyes. Optic capture was not necessary to ensure a clear visual axis. Primary posterior capsulotomy should be performed in preschool and uncooperative children and in eyes expected to have relatively high postoperative inflammation. Implanting the AcrySof in the bag and leaving the posterior capsule intact is acceptable for school children and juveniles with isolated developmental cataract. PMID- 12648641 TI - Comparison of secondary implantation of flexible open-loop anterior chamber and scleral-fixated posterior chamber intraocular lenses. AB - PURPOSE: To analyze and compare the outcomes and complication rates of secondary implantation of flexible, open-loop, anterior chamber intraocular lenses (AC IOLs) and single-piece, scleral-fixated, posterior chamber IOLs (PC IOLs). SETTING: Departments of Ophthalmology, Gaziantep University Medical Faculty, Research Hospital, Gaziantep, and Inonu University Medical Faculty, Research Hospital, Malatya, Turkey. METHODS: This study comprised 124 eyes of 113 aphakic patients (61 men, 52 women) with insufficient capsule support who had secondary IOL implantation from January 1997 to June 2001. In Group 1 (n = 73 eyes), a flexible, open-loop AC IOL was implanted and in Group 2 (n = 51 eyes), a single piece, scleral-fixated PC IOL. The mean follow-up was 34 months (range 6 to 53 months) and took place at several different clinical settings. The mean interval between the initial cataract operation and secondary IOL implantation was 63 months (range 6 months to 12 years). The postoperative outcomes, safety, efficacy, and complication rates were analyzed, and the preoperative and postoperative best spectacle-corrected visual acuities (BSCVAs) were compared. RESULTS: The postoperative mean BSCVA was 20/34.8 +/- 45.2 (SD) in Group 1 and 20/32.1 +/- 33.7 in Group 2; the difference was not significant (P =.718). A BSCVA of 20/40 or better was achieved in 62 eyes (84.9%) in Group 1 and 45 eyes (88.2%) in Group 2. A BSCVA of 20/25 or better was achieved in 40 eyes (54.8%) and 33 eyes (64.7%), respectively. The difference between the 2 groups was not significant (P =.472). Complications occurred in 25 eyes (34.2%) in Group 1 and 13 eyes (25.5%) in Group 2 (P >.05). The most frequent complications in Group 1 were early transient corneal edema, intraocular pressure elevation, cystoid macular edema, hyphema, secondary glaucoma, and iris capture or pupil decentration and in Group 2, suture erosion, a tilted or decentered IOL, fibrin reaction, and vitreous prolapse into the anterior chamber. CONCLUSIONS: The AC IOLs and PC IOLs were safe and effective for secondary implantation to correct aphakia. Secondary implantation of the scleral-fixated PC IOL seemed to provide a more favorable outcome and a lower complication rate than the open-loop AC IOL in complicated cataract cases with inadequate capsule and zonular support. As scleral-fixated PC IOL implantation is technically more difficult than AC IOL implantation, the decisive factor in choosing a secondary IOL is surgical experience. Long-term comparison of both techniques is required. PMID- 12648642 TI - Corticocapsular adhesions and their effect during cataract surgery. AB - PURPOSE: To identify the types of corticocapsular adhesions by their preoperative clinical appearance and evaluate the difficulties they pose during cataract surgery. SETTING: Iladevi Cataract & IOL Research Center, Ahmedabad, India. METHODS: Seventy-six eyes with age-related senile cataract scheduled for phacoemulsification were identified as having corticocapsular adhesions. Preoperatively, the eyes were assessed at the slitlamp to determine the type of corticocapsular adhesions present. During surgery, the surgeon graded nucleus rotation after single-site cortical-cleaving hydrodissection as easy, difficult, or not possible. Additional multiquadrant-focal hydrodissection was performed. The surgeon's impressions of the presence of corticocapsular adhesions and visualization of the furry surface of the epinucleus were noted. RESULTS: Corticocapsular adhesions were confirmed in 86.84% of eyes. Equatorial corticocapsular adhesions alone or in combination were present in 72 eyes (94.74%) eyes, anterior corticocapsular adhesions in 40 (52.74%), and posterior corticocapsular adhesions in 42 (56.26%). Rotation of nucleus was not possible in 47.37% eyes, difficult in 39.47%, and easy in 13.16%. CONCLUSIONS: The surgeon should perform a thorough preoperative slitlamp evaluation in extreme gaze with a fully dilated pupil. Equatorial corticocapsular adhesions, which were present in most eyes, made nucleus rotation difficult. Additional multiquadrant and focal cortical-cleaving hydrodissection helped separate the adhesions and achieve successful rotation. PMID- 12648643 TI - Efficacy of a capsular tension ring for phacoemulsification in eyes with zonular dialysis. AB - PURPOSE: To determine the safety and efficacy of capsular tension ring (CTR) insertion in eyes with zonular dialysis of less than 150 degrees having phacoemulsification with posterior chamber intraocular lens (PC IOL) implantation. SETTING: Dr. Agarwal's Eye Hospital, Chennai, India. METHODS: This prospective study comprised 21 eyes of 19 patients with zonular dialysis of less than approximately 150 degrees determined preoperatively or intraoperatively. After insertion of a CTR, phacoemulsification with PC IOL implantation was performed. The mean follow-up was 242.33 days. RESULTS: Capsule collapse did not occur in any eye with a CTR. Intraoperative extension of the dialysis occurred in 2 eyes (9.52%). The IOL was placed in the bag in all the eyes except 1 that had traumatic cataract and received a scleral-fixated IOL during a subsequent surgery. Postoperatively, there was minimal corneal edema in 2 eyes (9.52%) and mild iritis in 5 eyes (23.80%). Three eyes (14.28%) developed raised intraocular pressure that responded well to medical therapy. The symptoms resolved in the 3 patients with preoperative diplopia. Fifteen eyes (71.42%) had a final visual acuity of 6/12 or better. Six eyes had a final best corrected visual acuity of worse than 6/12 because of coexisting fundus pathology. A dilated pupil examination at 6 months showed a well-centered IOL in all eyes. CONCLUSIONS: Phacoemulsification with in-the-bag PC IOL and CTR implantation in eyes with zonular dialysis of up to approximately 150 degrees had a success rate of 90.47%. Visual recovery was not as good as in normal eyes because of the problems associated with zonular dialysis. PMID- 12648644 TI - Intrastromal corneal ring implantation for the correction of myopia: 12-month follow-up. AB - PURPOSE: To evaluate the efficacy, predictability, and stability of refraction obtained after intrastromal corneal ring segment (ICRS) implantation for low to moderate myopia. SETTING: Single-center clinical practice. METHODS: In this prospective 2-surgeon study, 9 patients (15 eyes) with low to moderate myopia were recruited to receive ICRS implants. RESULTS: At 1 day, 10 of the 15 eyes had an uncorrected visual acuity (UCVA) of 20/40 or better. At 12 months, all eyes had this UCVA and 66.6% had 20/25 or better. The mean manifest refraction stabilized after the first week at <-0.5 diopter (D). At 12 months, all eyes were within +/-1.0 D of the intended manifest refraction; 67% were within +/-0.5 D. Sixty percent of eyes had no change from the preoperative best corrected visual acuity; 13.3% improved by 1 line, and 26.6% lost 1 line. The postoperative complications included lamellar channel deposits (n = 12), ICRS dislocation (n = 2), corneal infiltrates (n = 2), bleeding in the positioning ring hole (n = 1), 0.3 mm segment decentration (n = 1), and prolonged wound healing (n = 1). CONCLUSIONS: Intrastromal corneal ring segment implantation for the correction of low to moderate myopia afforded good visual recovery and efficacy similar to that with laser in situ keratomileusis and superior to that with photorefractive keratectomy. However, light or blunt trauma and insufficient hygiene can have serious consequences and there is the potential for induced astigmatism. Corneal infiltrates can occur and must be treated immediately. The ring implantation technique is demanding. Advantages of ICRS implantation include rapid and stable visual recovery as well as reversibility. PMID- 12648645 TI - Advanced glycation end-products in anterior chamber aqueous of cataractous patients. AB - PURPOSE: To investigate whether the advanced glycation end-products (AGEs) pentosidine, N( epsilon )-(carboxymethyl)lysine (CML), and imidazolone are present in the aqueous of cataract patients and how AGE levels correlate to cataract type or the diabetic condition of the patient. SETTING: Departments of Ophthalmology and Internal Medicine, University of Jena, Jena, Germany. METHODS: Aqueous and serum samples from 77 cataractous patients (33 nondiabetics, 44 diabetics; 14 with dense posterior, 63 with nuclear cataracts) were investigated. The mean age of the patients was 69 years +/- 14 (SD). The aqueous protein concentration was examined using a laser flare-cell meter. In the samples, pentosidine was measured by high-performance liquid chromatography and CML using a competitive enzyme-linked immunosorbent assay. Western blot analysis was used to detect imidazolone, pentosidine, and CML in the aqueous. RESULTS: The aqueous samples contained CML, pentosidine, or imidazolone. These AGEs occurred mainly bound to albumin. Significant correlations existed between serum pentosidine and aqueous CML and flare levels as well as between serum and aqueous CML. Patients with nuclear cataract had insignificantly higher pentosidine and CML levels than patients with posterior cataract, whereas the flare was significantly higher. No significant differences were found between the aqueous AGE levels in nondiabetic and diabetic patients. CONCLUSIONS: The aqueous of cataractous eyes contained the AGEs CML, pentosidine, and imidazolone. All 3 AGEs occurred mainly albumin-bound, providing evidence they may originate from the blood. Further investigation is needed to determine the relevance of aqueous AGEs in cataractogenesis. PMID- 12648646 TI - Intraocular lens surfaces and their relationship to postoperative glare. AB - PURPOSE: To estimate the potential for surface reflections in recently introduced intraocular lenses (IOLs) and to determine optic surface designs that will reduce surface reflections. SETTING: Mayo Clinic, Rochester, Minnesota, USA. METHODS: Surface-reflected glare in the unaccommodated human crystalline lens and in 6 IOLs (Bausch & Lomb SoFlex LI61U, Pharmacia CeeOn 911A, Allergan Sensar AR40, Bausch & Lomb Hydroview H60M, Alcon AcrySof MA60BM, Alcon AcrySof SA30AL) was examined in a physiologic eye model using the ZEMAX optical design program. Internal and external surface reflections were described and compared in terms of IOL surface reflectivity (%), area of the reflected glare image (mm(2)), and relative intensity of the reflected glare image. RESULTS: Compared to surface reflections from the unaccommodated human lens with a corneal power of 43.0 diopters, all the IOLs increased the relative intensity of internal and external reflections by 3- to 36-fold except the MA60BM and the SA30AL, which increased the relative intensity of internal and external reflections by 730- to 1090-fold. CONCLUSIONS: All the IOLs studied variably increased internal and external surface reflections when compared to the human crystalline lens. Surface reflections were minimized in optic designs with an anterior radius of curvature of approximately 17.0 mm or less. PMID- 12648647 TI - Computerized analysis of the effects of intraocular lens edge design on the quality of vision in pseudophakic patients. AB - PURPOSE: To study the distribution and intensity of reflected glare images from 3 intraocular lens (IOL) edge designs. SETTING: Eye Institute, University of Florence, Florence, Italy. METHODS: The interaction of light rays as a function of edge design was evaluated using an experimental eye model and an advanced ray tracing program. Three hydrophobic acrylic IOLs were studied: an acrylic square edged (sharp) design, an acrylic round-edged design, and a new acrylic IOL with a rounded anterior edge and a sharp posterior edge. RESULTS: The rays hitting the sharp edge of an IOL caused peripheral arcs or circles of light on the side of the retina opposite the focused image. Edge reflections also occurred with the round-edged IOL, but the intensity of the reflected image was one tenth that of the intensity with the sharp-edged design. The new round-/sharp-edged design reduced the maximum intensity of the reflected image to one third the intensity of the sharp-edged reflected image. CONCLUSIONS: The experimental findings with an acrylic IOL that combines a sharp posterior edge with a low incidence of reflected edge glare are promising. PMID- 12648648 TI - Effect of the optic size of a single-piece acrylic intraocular lens on posterior capsule opacification. AB - PURPOSE: To investigate the effect of the optic size of an intraocular lens (IOL) with sharp optic edges on posterior capsule opacification (PCO). SETTING: Nishi Eye Hospital, Osaka, Japan. METHODS: In 5 rabbits, a single-piece 5.5 mm optic AcrySof IOL (Alcon Laboratories) was implanted in 1 eye and a specially fabricated single-piece 7.0 mm optic AcrySof IOL was implanted in the contralateral eye. Histopathological examinations were performed 3 weeks after surgery. RESULTS: On posterior views, less PCO was noted with the 5.5 mm optic in all rabbits except 1. Histopathological sections demonstrated adhesion of the anterior and posterior capsules between the haptic and optic and the formation of a sharp capsular bend at the posterior optic edge in 3 eyes with the 5.5 mm optic but in no eye with the 7.0 mm optic. Abundant PCO was noted when a sharp capsular bend had not formed. CONCLUSIONS: Capsular adhesion is a prerequisite of capsular bend formation. The sharp optic edge alone does not provide a substantial barrier when a capsular bend is not formed. Bulky haptics such as those of the single piece AcrySof and large optics may hamper capsular adhesion and bend formation. PMID- 12648649 TI - Effect of tenascin and fibronectin on the migration of human corneal fibroblasts. AB - PURPOSE: To investigate the effect of fibronectin and tenascin on the migration of corneal fibroblasts. SETTING: Department of Ophthalmology, University of Vienna, Medical School, Vienna, Austria. METHODS: Using human corneal fibroblasts, a monolayer migration assay was performed to measure corneal fibroblast movement. The migration on collagen I, fibronectin, and tenascin with and without transforming growth factor (TGF)-alpha/fibroblast growth factor (FGF) 2 stimulation and the effect of soluble tenascin and fibronectin on corneal fibroblast migration on collagen-I-coated wells were investigated. RESULTS: The cytokines TGF-alpha and FGF-2 stimulated migrational activity of corneal stromal cells in a dose-dependent manner, reaching the maximum effect at 100 ng/mL and 10 ng/mL, respectively. The migration of corneal fibroblasts on fibronectin was significantly higher (P <.05) than the migration on collagen I. Transforming growth factor-alpha and FGF-2 increased radial cell displacement independent of the provided matrix composition. Tenascin had a negative effect on corneal fibroblast adhesion/migration in this in vitro model. CONCLUSION: Fibronectin and tenascin influenced corneal fibroblast migration and adhesion, respectively, and may play a role in stromal cell movement during wound healing. The cytokines TGF alpha and FGF-2 had an additive effect on corneal fibroblast migration on a fibronectin matrix. PMID- 12648650 TI - Bacterial adhesion to rigid and foldable posterior chamber intraocular lenses: in vitro study. AB - PURPOSE: To determine the in vitro bacterial adherence to intraocular lenses (IOLs) routinely used at 1 center. SETTING: University Hospital of Vienna, Vienna, Austria. METHODS: Four types of rigid IOLs (hydrophobic poly[methyl methacrylate] [PMMA] and hydrophilic heparin-surface-modified PMMA) and 5 types of foldable intraocular lenses (hydrophobic silicone, hydrophobic acrylic, and hydrophilic acrylic) were used in the experiment. Under standardized conditions, the IOLs were contaminated with 2 strains of Staphylococcus epidermidis. A sonication method and impression method were used for quantification. The mean bacterial density per lens type (colony forming units/mm(2)) was compared. RESULTS: With both germs and both quantification methods, more bacteria was found on hydrophobic lenses than on IOLs with hydrophilic surfaces (P =.001). The Alcon AcrySof and Askin UV80F IOLs had the greatest and the Corneal Acrygel and Bausch & Lomb Hydroview IOLs the least affinity to these microorganisms. CONCLUSION: The data suggest that hydrophilic IOLs can help reduce the rate of postoperative endophthalmitis as a result of their surface properties. PMID- 12648651 TI - Surface roughness of intraocular lenses and inflammatory cell adhesion to lens surfaces. AB - PURPOSE: To assess and examine the relationship between the degree of intraocular lens (IOL) surface roughness and the severity of inflammatory cell adhesion to the optical surface. SETTING: Department of Ophthalmology, Tokyo Medical University Hospital, Tokyo, Japan. METHODS: Poly(methyl methacrylate) IOLs with 3 degrees of mean surface roughness (Ra) were used including IOLs with an unpolished optical surface and IOLs with optical surfaces polished for 1 day or 5 days. The IOLs were cultured for 96 hours with 1 cell/mL x 10(7) cells/mL of splenocytes from rats that developed experimental autoimmune uveoretinitis (EAU). At the end of culture, the number of inflammatory cells adhering to the optical surfaces of the IOLs was counted. RESULTS: The mean Ra for the unpolished IOLs, IOLs polished for 1 day, and IOLs polished for 5 days was 14.17 nm, 3.71 nm, and 1.37 nm, respectively. The mean numbers of cells that adhered to the optical surfaces of the IOLs after culturing with splenocytes from rats with EAU were 86 cells/mm(2), 66 cells/mm(2), and 48 cells/mm(2), respectively. There was a significant difference between the unpolished IOLs and the IOLs polished for 5 days (P =.003, Mann-Whitney U test). CONCLUSION: Inflammatory cell adhesion to the IOL optical surface was affected by the Ra value of the IOL. PMID- 12648652 TI - In vitro study of bacterial adherence to processed dura mater, processed pericardium, pericardium in saline, and human sclera. AB - PURPOSE: To study bacterial adherence to processed dura mater, processed pericardium, pericardium in saline, and human sclera and the difference in bacterial adherence to these tissues. SETTING: Research Laboratory, Loyola University Medical Center, Maywood, Illinois, USA. METHODS: Specimens of processed dura mater, processed human pericardium, pericardium in saline, and human sclera (N = 32) were exposed to Staphylococcus epidermidis (concentration 3 x 10(8)) for 10, 20, 40, and 60 minutes, washed for 5 seconds, fixed, and processed for scanning electron microscopy (SEM). Each bacterial count represents an average of 50 random SEM fields at x5,000 magnification. After SEM, selected specimens were processed for transmission electron microscopy. RESULTS: The mean number of bacteria/mm(2) +/- SD adhering to the tissues at 10, 20, 40, and 60 minutes, respectively, were dura mater, 107,833 +/- 65,410, 104,500 +/- 13,471, 96,067 +/- 113,884, and 204,267 +/- 153,697; processed pericardium, 131,550 +/- 86,194, 100,900 +/- 20,031, 144,683 +/- 51,730, and 176,933 +/- 111,818; pericardium in saline, 7,925 +/- 1,520, 33,933 +/- 32,085, 1,217 +/- 1,287, and 21,550 +/- 16,436; and human sclera, 4,850 +/- 2,121, 23,700 +/- 17,961, 5,150 +/ 1,273, and 8,175 +/- 8,450. A 2-way analysis of variance showed significant differences among groups (P =.001) and no significant difference in sample time (P =.929). CONCLUSIONS: Bacterial adherence to processed dura mater, processed pericardium, pericardium in saline, and human sclera should be considered when these materials are used clinically during ophthalmic surgery and other surgical specialties. Adequate broad-spectrum antibiotic coverage is needed to prevent infection and subsequent complications in patients. PMID- 12648653 TI - Current surgical options for keratoconus. AB - The treatment of keratoconus is partly dependent on the severity of the disease process. When contact lenses fail because the patient is intolerant or they do not provide sufficient acuity to meet the patient's needs, penetrating keratoplasty is the usual surgical modality for adequate visual rehabilitation. Keratoconus is considered a contraindication to incisional refractive surgery such as radial keratotomy and laser in situ keratomileusis because of the corneal instability conferred by these procedures. When the cornea is transparent, intrastromal corneal ring implantation is a refractive modality that may improve visual function and, in select keratoconus patients, obviate the need for corneal transplantation. PMID- 12648654 TI - Aberrant regeneration of corneal nerves after laser in situ keratomileusis. AB - We report a case of aberrant regeneration of corneal nerves along the corneal flap interface after myopic laser in situ keratomileusis (LASIK) using confocal microscopy in vivo. The aberrant stromal nerves persisted at the last follow-up, 2 years post LASIK. The short-term clinical outcome was excellent. The long-term clinical effects are unknown. PMID- 12648656 TI - Tissue adhesive to treat 2-site corneal melting associated with topical ketorolac use. AB - We report a case of a 78-year-old man presenting with 2 discrete areas of sterile corneal melting associated with chronic use of topical ketorolac after uneventful clear corneal phacoemulsification. He was treated successfully with tissue adhesive application. Patients receiving chronic topical ketorolac treatment, especially those with ocular surface abnormalities, can present with severe complications such as corneal melting. PMID- 12648655 TI - Prophylactic use of mitomycin-C in the management of a buttonholed LASIK flap. AB - We report a case in which prophylactic mitomycin-C (MMC) was used to prevent haze and scarring after photorefractive keratectomy (PRK) in an eye with a previous laser in situ keratomileusis flap buttonhole. During the patient's primary procedure, a 3.5 mm central buttonhole was created during the microkeratome pass. The flap was repositioned without laser application. Follow-up examinations revealed a moderate central buttonhole scar. Twelve months later, the patient had PRK with adjunctive MMC 0.02% application in the eye. At the 1-year examination, the uncorrected visual acuity was 20/20 and the cornea was clear with no evidence of scarring or haze. PMID- 12648657 TI - Corneal wedge resection to treat progressive keratoconus in the host cornea after penetrating keratoplasty. AB - We present 3 cases of progressive keratoconus in the host after penetrating keratoplasty PKP that were treated with a wide crescentic excision of the diseased tissue in the host cornea guided by Orbscan II topography (Bausch & Lomb). In all cases, the procedure resulted in a more regular corneal shape with a marked reduction in astigmatism, preventing the need for repeat PKP PMID- 12648658 TI - Sutured piggyback toric intraocular lenses to correct high astigmatism. AB - A 74-year-old woman presented for bilateral cataract surgery, which was performed 3 days apart. Corneal astigmatism was 4.25 diopters (D) in the right eye and 4.87 D in the left. After cataract extraction through 6.0 mm scleral incisions, 2 toric, plate-haptic, silicone intraocular lenses (IOLs), each with a 3.50 D cylinder add power (2.30 D at spectacle plane), were sutured together and implanted in the bag. Both eyes had limbal relaxing incisions postoperatively. Four months postoperatively, corneal astigmatism was 4.50 D in the right eye and 4.00 D in the left. Refractive astigmatism was 0.50 D with an uncorrected visual acuity of 20/40 in both eyes. No IOL rotation was observed. Suturing toric lenses together allows greater correction of astigmatism without concern about counter rotation of the lenses. PMID- 12648659 TI - Congenital double crystalline lenses. AB - We report an unusual case of an eye with 2 congenital crystalline lenses; ie, a duplicated lens. The horizontal axis of the eye was obliquely placed. One lens was situated in the upper temporal quadrant and the other, in the lower nasal quadrant. One lens was larger than the other. PMID- 12648660 TI - Late bacterial keratitis after implantation of intrastromal corneal ring segments. AB - We report a case of bacterial keratitis that occurred after implantation of intrastromal corneal ring segments (Intacs). The patient presented with decreased vision, inflammation, and stromal infiltrates localized at the extremity of an Intacs channel 3 months after surgery. Culture were positive for Clostridium perfringens and Staphylococcus epidermidis. The infiltrates progressed despite treatment with topical fortified and systemic antibiotics. The Intacs were removed. The keratitis slowly resolved, and the patient recovered a best corrected visual acuity of 20/20. PMID- 12648661 TI - Spontaneous posterior dislocation of an intraocular lens presenting as pupillary block glaucoma. AB - We describe spontaneous posterior dislocation of an in-the-bag, plate-haptic silicone intraocular lens (IOL) 1 year after neodymium:YAG laser posterior capsulotomy. The patient presented with an acutely painful eye associated with loss of vision. The initial cataract surgery was uneventful. Clinical findings at presentation were suggestive of pupillary block glaucoma attributable to vitreous herniation into the anterior chamber. The acute attack was broken after pupil dilation. The patient subsequently had removal of the dislocated IOL via a pars plana vitrectomy and implantation of an IOL in the ciliary sulcus. PMID- 12648663 TI - Delayed opacification of a PMMA intraocular lens. PMID- 12648662 TI - Using nylon hooks during small-pupil phacoemulsification. PMID- 12648664 TI - Induced astigmatism after laser in situ keratomileusis. PMID- 12648665 TI - Recurrent capsule opacity and erythropoietin. PMID- 12648667 TI - Negative receptor signalling. AB - Binding of external factors to cell membrane receptors triggers intracellular signalling pathways that ultimately determine if the cell proliferates, differentiates or undergoes apoptosis. Activated receptors also initiate a cascade of events, called negative receptor signalling, that decreases the amplitude of positive signals and modulates the level of cell stimulation. Recent studies have revealed that negative signalling by receptor tyrosine kinases involves coordinated action of ubiquitin ligases (i.e. Cbl), adaptor proteins (i.e. Grb2 and CIN85), inhibitory molecules (i.e. Sprouty), cytoplasmic kinases (i.e. activated Cdc42-associated kinase) and phosphoinositol metabolites. These inhibitory signals are essential for normal cell functioning, and their deregulation often results in human diseases. PMID- 12648668 TI - Ras pathway signaling on endomembranes. AB - Until recently, the plasma membrane has been considered to be a unique platform from which emanate the signaling events regulating or regulated by Ras and its close relatives. For the past few years, the role of endosomes derived from the plasma membrane as platforms for Ras/mitogen-activated protein kinase signaling has been appreciated. More recently, the cytoplasmic face of the Golgi apparatus and endoplasmic reticulum have been shown to host Ras signaling. The biological implications of compartmentalized signaling are only beginning to emerge. PMID- 12648669 TI - Nuclear localization and possible functions of receptor tyrosine kinases. AB - Recent data have renewed interest in the possible nuclear localization of receptor tyrosine kinases, as well as their ligands. In one case, that of ErbB-4, the receptor is processed by two membrane-localized proteases to produce a soluble cytoplasmic domain fragment that includes the tyrosine kinase domain. This fragment, generated by a metalloprotease-dependent ectodomain cleavage followed by gamma-secretase cleavage within the transmembrane domain, is also found in the nucleus. Three other receptor tyrosine kinases have been detected in the nucleus in the absence of proteolytic processing. In some instances, nuclear localization of receptor tyrosine kinases is growth-factor-dependent and tentative evidence suggests a role in transcription. PMID- 12648670 TI - Nuclear choreography: interpretations from living cells. AB - The advent of green fluorescent protein technology, its use in photobleaching experiments and the development of methods to rapidly acquire images and analyze complex datasets have opened the door to unraveling the mechanisms of nuclear functions in living cells. Studies over the past few years have characterized the movement of chromatin, nuclear proteins and nuclear bodies and, in some cases, correlated their dynamics with energy dependence, cell cycle progression, developmental changes, factor targeting and nuclear position. The mechanisms by which nuclear components move or are restrained have important implications for understanding not only the efficacy of nuclear functions but also the regulation of developmental programs and cellular growth. PMID- 12648671 TI - Cyclin D1 serves as a cell cycle regulatory switch in actively proliferating cells. AB - Much of our current understanding of the cell cycle involves analyses of its induction in quiescent cells. To better understand the control of cell cycle propagation and termination, studies have been performed in actively cycling cultures using time-lapse photography and quantitative image analysis. These studies reveal a highly ordered sequence of events required for promotion of continued proliferation. The decision to continue cell cycle progression takes place in G2 phase, when cellular Ras induces the elevation of cyclin D1 levels. These levels are maintained through G1 phase and are required for the initiation of S phase, at which time cyclin D1 levels are automatically reduced to low levels. The reduction of cyclin D1 to low levels during S phase is required for DNA synthesis, and forces the cell to induce high cyclin D1 levels once again when it enters G2 phase. In this way, cyclin D1 is proposed to serve as an active switch in the regulation of continued cell cycle progression. PMID- 12648672 TI - Ubiquitination, phosphorylation and acetylation: the molecular basis for p53 regulation. AB - The p53 tumor suppressor exerts anti-proliferative effects, including growth arrest, apoptosis and cell senescence, in response to various types of stress. Tight regulation of p53 activation is imperative for preventing tumorigenesis and maintaining normal cell growth; p53 stabilization and transcriptional activation are crucial early events in a cell's battle against genotoxic stress. Ubiquitination, phosphorylation and acetylation are post-translational modifications to p53 that affect its overall appearance and activity. Recent findings suggest that these modifications have a profound affect on p53 stability and function. Defining the precise roles of these modifications in p53 function may show not only that they are markers of the stress response but also that they serve as the focal point in the regulation of p53. PMID- 12648673 TI - Histone and chromatin cross-talk. AB - Chromatin is the physiologically relevant substrate for all genetic processes inside the nuclei of eukaryotic cells. Dynamic changes in the local and global organization of chromatin are emerging as key regulators of genomic function. Indeed, a multitude of signals from outside and inside the cell converges on this gigantic signaling platform. Numerous post-translational modifications of histones, the main protein components of chromatin, have been documented and analyzed in detail. These 'marks' appear to crucially mediate the functional activity of the genome in response to upstream signaling pathways. Different layers of cross-talk between several components of this complex regulatory system are emerging, and these epigenetic circuits are the focus of this review. PMID- 12648674 TI - Ubiquitin: not just for proteasomes anymore. AB - Ubiquitin is a small protein that can be covalently linked to itself or other proteins, either as single ubiquitin molecules or as chains of polyubiquitin. Addition of ubiquitin to a target protein requires a series of enzymatic activities (by ubiquitin-activating, -conjugating and -ligating enzymes). The first function attributed to ubiquitin was the covalent modification of misfolded cytoplasmic proteins, thereby directing proteasome-dependent proteolysis. More recently, additional functions have been ascribed to ubiquitin and ubiquitin related proteins. Ubiquitin directs specific proteins through the endocytic pathway by modifying cargo proteins, and possibly also components of the cytoplasmic protein trafficking machinery. PMID- 12648675 TI - Function prediction and protein networks. AB - In the genomics era, the interactions between proteins are at the center of attention. Genomic-context methods used to predict these interactions have been put on a quantitative basis, revealing that they are at least on an equal footing with genomics experimental data. A survey of experimentally confirmed predictions proves the applicability of these methods, and new concepts to predict protein interactions in eukaryotes have been described. Finally, the interaction networks that can be obtained by combining the predicted pair-wise interactions have enough internal structure to detect higher levels of organization, such as 'functional modules'. PMID- 12648676 TI - Global approaches to protein-protein interactions. AB - The availability of complete, annotated genome sequences for a variety of eukaryotic organisms has paved the way for a paradigm shift in biomedical research from the 'one gene-one hypothesis' approach to more global, systematic strategies that analyse genes or proteins on a genome- and proteome-wide scale. One daunting task in the post-genome era is to determine how the complement of expressed cellular proteins - the proteome - is organised into functional, higher order networks, by mapping all constitutive and dynamic protein-protein interactions. Traditionally, reductionist approaches have typically focused on a few, selected gene products and their interactions in a particular physiological context. In contrast, more holistic strategies aim at understanding complex biological systems, for example global protein-protein interaction networks on a cellular or organismal level. Several large-scale proteomics technologies have been developed to generate comprehensive, cellular protein-protein interaction maps. PMID- 12648677 TI - Functional genomic maps in Caenorhabditis elegans. AB - The completion of the Caenorhabditis elegans genome sequence was the initial step toward the use of whole-genome analysis in this model organism. Advances in C. elegans genomics include transcript profiling, gene-function screens using RNA mediated interference, and protein-interaction mapping using the yeast two-hybrid system. Recent reports have employed these methods to gain new insights into diverse biological problems such as tissue-specific gene expression, cell-fate specification, genome organization, the DNA damage response, and early embryonic development. These studies combined genomic approaches to probe complex biological pathways on an unprecedented scale. PMID- 12648678 TI - Profiling cancer. AB - In the past couple of years, several very exciting studies have demonstrated the enormous power of gene-expression profiling for cancer classification and prediction of patient survival. In addition to promising a more accurate classification of cancer and therefore better treatment of patients, gene expression profiling can result in the identification of novel potential targets for cancer therapy and a better understanding of the molecular mechanisms leading to cancer. PMID- 12648679 TI - Sniffers, buzzers, toggles and blinkers: dynamics of regulatory and signaling pathways in the cell. AB - The physiological responses of cells to external and internal stimuli are governed by genes and proteins interacting in complex networks whose dynamical properties are impossible to understand by intuitive reasoning alone. Recent advances by theoretical biologists have demonstrated that molecular regulatory networks can be accurately modeled in mathematical terms. These models shed light on the design principles of biological control systems and make predictions that have been verified experimentally. PMID- 12648680 TI - Divide and conquer: nucleotide excision repair battles cancer and ageing. AB - Protection from cancer and ensured longevity are tightly linked in mammals. One of the fundamental mechanisms contributing to both is the cellular response to DNA damage. The appropriate response is an initial attempt at repair, but if the damage is too extensive or compromises DNA metabolism, a signalling cascade triggers cellular senescence or death. Evidence in mice and humans suggests a division of tasks amongst DNA repair pathways: transcription-coupled repair and interstrand crosslink repair of cytotoxic lesions are predominantly responsible for longevity assurance, whereas excision repair of mutagenic lesions provides protection against cancer. Similarly, the signalling component of the DNA-damage response might contribute unequally to organismal outcomes depending on its set point: an inadequate response to DNA damage sanctions carcinogenesis but might limit local ageing, whereas overzealous signalling provides cancer protection but accelerates ageing. PMID- 12648681 TI - Nicotinamide adenine dinucleotide, a metabolic regulator of transcription, longevity and disease. AB - Nicotinamide adenine dinucleotide (NAD) is a ubiquitous biological molecule that participates in many metabolic reactions. Recent studies show that NAD also plays important roles in transcriptional regulation, longevity, calorie-restriction mediated life-span extension and age-associated diseases. It has been shown that NAD affects longevity and transcriptional silencing through the regulation of the Sir2p family, which are NAD-dependent deacetylases. Many human diseases are associated with changes in NAD level and/or the NAD : NADH ratio, raising the possibility that the Sir2p family might play a role in these diseases. PMID- 12648682 TI - Oxidant signals and oxidative stress. AB - Although oxidants clearly possess the capacity to behave in a random and destructive fashion, growing evidence suggests that in many instances the production of reactive oxygen species is tightly regulated and their downstream targets exquisitely specific. This past year, several notable advances have been made in defining the specific redox-dependent targets of intracellular oxidants, as well as the myriad pathways that appear to employ oxidants as effector molecules. These new studies have significantly altered our understanding of how reactive oxygen species participate in diverse processes from tumourigenesis to ageing. PMID- 12648683 TI - Esophageal cancer: what price swallowing? PMID- 12648685 TI - Octreotide in the treatment of lymphorrhea after axillary node dissection: a prospective randomized controlled trial. AB - BACKGROUND: Axillary lymph node dissection for staging and local control of nodal disease is an integral part of breast cancer therapy. Lymphorrea is a serious and disabling complication of axillary lymphadenectomy, but no effective therapy is currently available. Octreotide is a hormone with general antisecretory effects that has been used to control lymphorrhea in thoracic duct injury and after radical neck dissection. The aim of the study we describe in this article was to determine whether octreotide has a role in the treatment of post axillary lymphadenectomy lymphorrhea. STUDY DESIGN: This is a prospective randomized controlled trial. Two hundred sixty-one consecutive patients with various stages of breast cancer who underwent axillary lymph node dissection were randomized and followed for 7 years. The treatment group received 0.1 mg octreotide subcutaneously three times a day for 5 days, starting on the first postoperative day, while the control group received no treatment. Of the 261 patients undergoing axillary node dissection, 136 were assigned to the control group and 125 composed the treatment group. The control group and the treatment group were evaluated for amount and duration of lymphorrhea as well as inflammatory and infectious complications. RESULTS: In the control group, the mean quantity (+/- standard deviation) of lymphorrhea was 94.6 +/- 19 cc per day and the average duration was 16.7 +/- 3.0 days. In comparison, the mean quantity of lymphorrhea in the treatment group was 65.4 +/- 21.1 cc (p < 0.0001) per day and the average duration was 7.1 +/- 2.9 days (p < 0.0001). We did not find an important difference in the number of infectious complication or hematomas formation between the study groups. CONCLUSIONS: Octreotide can be used successfully for the treatment of post-axillary dissection lymphorrea, and potentially, in the prevention of post-axillary lymph node dissection lymphosarcoma, since the amount and duration of lymphorrhea in this setting are known to be important risk factors for its development. Potentially, octreotide might be used in similar situations where lymphorrhea is detrimental, such as peripheral vascular surgery and regional lymph node dissection for melanoma. PMID- 12648684 TI - Surgical decision making and factors determining a diagnosis of breast carcinoma in women presenting with nipple discharge. AB - BACKGROUND: There is no consensus about the use of the various diagnostic tests and surgical procedures available to confirm or rule out breast cancer in patients presenting with nipple discharge. This study was designed to identify patient and nipple-discharge characteristics associated with the diagnosis of breast cancer and to determine the utility of mammography, sonography, ductography, and cytology in surgical decision making in patients presenting with pathologic nipple discharge. STUDY DESIGN: We reviewed the medical records of all patients who presented with nipple discharge at our institution between August 1993 and September 2000. Patient and nipple-discharge characteristics and findings on imaging studies and cytologic examination were analyzed. RESULTS: A total of 146 patients presented at our institution with nipple discharge during the study period. Of these, 52 had clinically benign discharge and were managed without surgical intervention; 94 patients had pathologic discharge and underwent a biopsy procedure for histologic diagnosis, treatment, or both. Logistic regression analysis identified mammographic (relative risk [RR] = 10.47, 95% confidence interval [CI] 2.36 to 46.39, p = 0.0002) and sonographic (RR = 5.54, 95% CI 1.27 to 25.40, p = 0.028) abnormalities as independent factors associated with a malignant diagnosis. Nineteen cancers, 62 papillomas, and 13 other benign lesions were identified among the patients with pathologic discharge. In 3 patients with cancer (15.8%) and 30 patients with a papilloma (48.4%), ductography was the only means of identifying lesions to be resected. Patients who underwent ductography-guided operation (n = 42, 50%) or any surgical procedure including a localization study (n = 66, 78.6%) were significantly more likely than patients who underwent central duct excision alone to have a specific underlying lesion identified (p = 0.045 and p = 0.033, respectively). CONCLUSIONS: Abnormalities on mammography and sonography in patients with nipple discharge should alert physicians to the possibility of a breast cancer diagnosis. In patients with pathologic discharge with normal findings on physical examination and other imaging studies, ductography might be the only means of localizing and resecting breast lesions associated with nipple discharge. PMID- 12648686 TI - Cricopharyngeal myotomy in the treatment of Zenker's diverticulum. AB - BACKGROUND: Transection of the cricopharyngeus muscle supposedly is of crucial therapeutic importance, because of its hypothesized pathogenic role in Zenker's diverticulum (ZD). This retrospective, nonrandomized study evaluates the outcomes of surgical therapy, with reference to cricopharyngeal myotomy (CM). STUDY DESIGN: Seventy-nine patients underwent diverticulectomy from 1985 to 1999. Group A (n = 47; men:women, 29:18; mean age +/- SD: 69 +/- 12 [range 35 to 87] years) underwent CM because of clearly discernible hypertrophic transverse fibers. In group B (n = 32; men:women, 22:10; mean age +/- SD 68 +/- 13 [range 36 to 95] years), without such transverse fibers, no CM was carried out. Dysphagia, regurgitation, and diverticular radiomorphology were classified according to ordinal scales. Diverticular volumes were calculated from barium swallows. Postoperative symptoms and outcomes were evaluated by questionnaires sent to the patients. RESULTS: The two groups did not differ significantly in severity of preoperative dysphagia and regurgitation, radiomorphology, or median diverticular volume. Both groups experienced postoperative alleviation of symptoms (p < 0.001), persisting slightly in 11 of 47 (23%) group A and 4 of 32 (13%) group B patients (p > 0.05). Of these, seven group A (64%) patients and three group B patients (75%) had additional upper gastrointestinal tract (GIT) diseases. Recurrent diverticula occurred in one patient and postoperative complications in five patients per group (p > 0.05). Diverticular volume and upper GIT comorbidities, but not CM, were significant risk factors for persistent symptoms in the multivariate analysis. CONCLUSIONS: CM has no significant influence on postoperative results, if carried out depending on the anatomic state and configuration of the cricopharyngeus muscle. This is suggested by the favorable outcomes of patients, with persistent symptoms being caused by factors other than CM. PMID- 12648687 TI - Guidelines for withholding or termination of resuscitation in prehospital traumatic cardiopulmonary arrest. PMID- 12648689 TI - A comparison of diet and exercise therapy versus laparoscopic Roux-en-Y gastric bypass surgery for morbid obesity: a decision analysis model. AB - BACKGROUND: In the absence of randomized controlled trials that directly compare medical versus surgical treatment of morbid obesity, decision analysis is a useful tool to help determine the optimal treatment strategy. Using decision analysis we simulated a trial comparing diet and exercise therapy to laparoscopic gastric bypass surgery to determine which approach resulted in longer life expectancy. STUDY DESIGN: A Markov decision analysis model was constructed to evaluate survival after laparoscopic Roux-en-Y gastric bypass surgery compared with a diet and exercise program for a 45-year-old woman with a body mass index (BMI) of 40 kg/m(2). Baseline mortality data were derived from published tables of vital statistics, and the relative risks of death associated with obesity (relative to normal weight) were taken from epidemiologic studies. We assumed that successful surgery resulted in a reduction of BMI to 30 kg/m(2). The baseline assumptions were: an operative mortality of 0.4%; a probability of weight loss after surgery of 80%; a rate of weight loss on a diet and exercise program, 20% at two years; a rate of regain of lost weight, 95% at two years; a relative risk of death for a BMI of 40 kg/m(2), 2.70; and a relative risk of death for a BMI of 30 kg/m(2), 1.51. RESULTS: The undiscounted life expectancy after surgery was 69.7 years compared with 67.3 years for a diet and exercise program (an absolute increase in life expectancy of 2.4 years, a relative increase in life expectancy of 10.8%). Sensitivity analyses assumed discounting at 5%/y, and showed that surgery was associated with a longer expectation of life when the risk of operative mortality was less than 10%, and when the probability of weight loss after surgery was greater than 4%. CONCLUSIONS: In a decision analysis model, laparoscopic gastric bypass surgery for morbid obesity was associated with a substantially longer survival than diet and exercise therapy. PMID- 12648690 TI - A cost-effectiveness analysis of intraoperative cholangiography in the prevention of bile duct injury during laparoscopic cholecystectomy. AB - BACKGROUND: Recent population-based studies have demonstrated that the use of intraoperative cholangiography (IOC) during laparoscopic cholecystectomy (LC) is associated with a decrease in the rate of common bile duct (CBD) injury. The cost implications of a management strategy involving routine IOC use have not been adequately evaluated. STUDY DESIGN: Decision analytic models were developed to analyze costs and benefits of routine IOC use during LC. The models were used to calculate the cost per life saved, cost per CBD injury avoided, and incremental cost of IOC when used routinely. Transition probabilities, costs, and outcomes were derived from published sources. Sensitivity analyses were used to account for uncertainty in these estimates. RESULTS: Using base-case estimates, management of patients undergoing LC with routine IOC would cost 100 dollars more per LC. Routine IOC would prevent 2.5 deaths for every 10,000 patients at a cost of 390,000 dollars per life saved (13,900 dollars per life year saved). The cost per CBD injury avoided with IOC use is 87,143 dollars. The cost per CBD injury avoided is less for procedures done in high-risk patients (approximately 8,000 dollars) or by less experienced surgeons (approximately 61,000 dollars). CONCLUSIONS: These models describe settings where the cost of IOC and the reduction in CBD injury rates make routine IOC use cost effective. Routine IOC use among less experienced surgeons and in high-risk operations is the most cost effective, but the cost implications of routine use for the general population should also be considered cost effective. PMID- 12648691 TI - Papillary carcinoma of the extrahepatic bile duct: characteristic features and implications in surgical treatment. AB - BACKGROUND: Papillary carcinoma of the extrahepatic bile duct presents clinically and histologically distinct features relevant to surgical decision-making. STUDY DESIGN: Serial sections of 15 specimens of resected papillary carcinoma of the bile duct were histologically examined to determine mode of spread, possibility of multicentric tumor origins, and coincidence with other neoplastic lesions. The presence of anomalous pancreaticobiliary ductal union was also investigated. These characteristics were considered with regard to surgical treatment. RESULTS: Three patients displaying pancreaticobiliary maljunction and one of three patients with a long common channel (> or = 8 mm) exhibited multicentric tumors. Eight patients (53%) demonstrated superficial spread along a mean length of 37.8 mm (range, 5 to 67 mm) of bile duct mucosa. Multicentric tumors developed synchronously in 4 patients, while metachronous tumors were identified in three patients displaying tumor histology similar to the primary lesions. Two of these three underwent successful repeated resection. Concomitant neoplastic lesions in the biliary tract were identified as mucosal dysplasia in four patients and cholangiocellular carcinoma of the liver in two. All tumors but one were removed via hepatic lobectomy or pancreatoduodenectomy, or both, resulting in a 5-year survival rate of 60%. CONCLUSIONS: Aggressive resection offers clear survival benefits for patients presenting with tumors displaying extensive superficial spread or multicentric origins. Closer attention should be paid to long common channels in relation to carcinogenesis of the bile duct, in addition to pancreaticobiliary maljunction. The risk of secondary tumor development remains, particularly in patients with pancreaticobiliary maljunction even after excision of the tumor-bearing extrahepatic bile duct. PMID- 12648692 TI - The use of fresh frozen plasma after major hepatic resection for colorectal metastasis: is there a standard for transfusion? AB - BACKGROUND: Major hepatic resection is indicated for selected patients with colorectal metastasis to the liver. Transfusion of fresh frozen plasma (FFP) might be required after major hepatectomy because of blood loss or coagulopathy, but there are no standard criteria for the use of FFP in this setting. METHODS: We identified 260 patients from our prospective database who underwent major (> or =3 Couinaud segments) hepatectomy between May 1997 and February 2001 for colorectal metastasis. FFP use was determined and tested for its relationship to clinical and pathologic factors. A survey on FFP use was sent to 12 other hepatobiliary centers worldwide. RESULTS: There were 142 (55%) men, 118 (45%) women, and the median age was 63 years. The most common hepatic resections performed were right lobectomy (37%) and extended right lobectomy (33%). There were 83 (32%) patients who received FFP. In these patients, a total of 405 units of FFP were administered with a median of 4 units. The majority of patients who received FFP were transfused within the first two postoperative days, while there were only five (2%) patients who initially received FFP beyond that time. FFP was administered for a median prothrombin time of 16.9. Only one (0.4%) patient required reoperation for bleeding. Right lobectomy and extended right lobectomy were found to predict FFP use on multivariate analysis. Postoperative complications did not correlate with FFP use. The criteria used for FFP administration at other major hepatobiliary centers were found to be variable. CONCLUSIONS: There is no universal standard for FFP use following major hepatic resection for colorectal metastasis. Our criterion of a prothrombin time of 16-18 seconds is conservative but results only rarely in reoperation for bleeding. Prospective evaluation of a higher threshold for FFP administration, such as an International Normal Ratio of 2.0, should be performed to better define the guidelines for FFP use in patients undergoing major hepatectomy who have normal underlying hepatic parenchyma. PMID- 12648693 TI - Effects of hospital volume on life expectancy after selected cancer operations in older adults: a decision analysis. AB - BACKGROUND: In addition to lower operative mortality, patients undergoing selected cancer operations at high volume centers have improved longterm survival. We sought to determine the overall effect of hospital volume on life expectancy after cancer surgery. STUDY DESIGN: We used a Markov decision analysis model to estimate life expectancy for patients undergoing resection for pancreatic, lung, or colon cancer. Model inputs included probabilities of operative mortality and longterm survival. For input data, we examined operative mortality (in-hospital or within 30 days) stratified by volume in over 400,000 patients undergoing resection for these three cancers using the national Medicare database (1994-1999). Risks of late mortality were abstracted from published studies (MEDLINE, 1966 to present) to model the effect of hospital volume on longterm survival. In analysis, we first calculated life expectancy for patients undergoing surgery at very low, low, medium, high, and very high volume hospitals. We then explored the effects of various regionalization strategies. RESULTS: Life expectancy increased steadily with hospital volume for all three cancers. Life expectancy after pancreatic cancer resection increased linearly with hospital volume: from 1.9 years at very low volume centers to 3.6 years at very high volume centers. For lung cancer, life expectancy ranged from 5.4 to 6.6 years. Increases in life expectancy for colon cancer were not as dramatic: from 6.8 at very low volume hospitals to 7.4 years at very high volume hospitals. Differences in life expectancy across volume strata were largely attributable to differences in longterm survival, not operative mortality. From a policy perspective, regionalizing surgery for colon cancer would produce the greatest overall life-expectancy gains, but it would require moving most patients. CONCLUSIONS: Patients aged 65 and older with pancreatic, lung, and colon cancer have substantially greater life expectancy after cancer resection at higher volume hospitals. Further work is needed to understand the mechanisms underlying differences in performance across hospitals in cancer care. PMID- 12648694 TI - Thoracoabdominal approach for right-sided hepatic resection for hepatocellular carcinoma. AB - BACKGROUND: Resection of a large hepatocellular carcinoma in the right liver or a small tumor located at the superior and posterior part of the right liver requires extensive hepatic mobilization. A thoracoabdominal approach might facilitate hepatic resection in such situations, but the safety and benefits of this approach remain unclear. STUDY DESIGN: A retrospective study based on a prospectively collected database of 488 patients was performed to evaluate the perioperative outcomes of right-sided hepatic resection for hepatocellular carcinoma using the thoracoabdominal approach (n = 92) in comparison with the conventional abdominal approach (n = 396). RESULTS: The two groups were comparable in age, comorbid illnesses, liver function, tumor size, and underlying cirrhosis. There were more extended right hepatectomies in the thoracoabdominal approach group than in the abdominal approach group, but the difference was not significant (33.7% versus 26.0%, p = 0.14). A significantly higher proportion of patients in the thoracoabdominal approach group did not require blood transfusion compared with the abdominal approach group (66.3% versus 54.8%, p = 0.04). The operating time for the former group was longer (median 450 min versus 360 min, p < 0.001). There were no significant differences in hospital stay (median 12.5 days versus 13.0 days, p = 0.82), overall morbidity (41.3% versus 38.6%, p = 0.64), or hospital mortality (4.3% versus 7.3%, p = 0.37) between the two groups. CONCLUSIONS: The thoracoabdominal approach is a safe operative approach that can facilitate resection of massive tumors in the right liver or tumors involving segments 7 and 8 without increased morbidity. PMID- 12648695 TI - The influence of gender in patients undergoing coronary artery bypass graft surgery: an eight-year prospective hospitalized cohort study. AB - BACKGROUND: We prospectively assess if there are any outcome differences between men and women undergoing coronary artery bypass graft surgery. STUDY DESIGN: This is an eight-year, prospective, hospitalization cohort study. Data were collected on 225 variables concurrently with admission. The main outcome was mortality. In addition, we evaluated 12 morbidity outcomes. To minimize confounding, we controlled for 18 other variables. RESULTS: We analyzed 5,324 patients (1,742 women; 3,582 men). Women were significantly older, with more African Americans and more Medicaid-insured (p < 0.05). Men had significantly higher body surface area, creatinine, tobacco history, prior myocardial infarctions, left ventricular hypertrophy, and cardiac pump time (p < 0.05). Women had significantly more diabetes, hypercholesterolemia, hypertension, previous cerebrovascular disease, urgency of the surgical procedure, fewer arterial grafts, and reduced functional capacity by the New York Heart Association classification (p < 0.05). Using multiple regression analysis there was no difference for mortality, positive bacterial cultures, ICU length of stay, arrhythmias, reoperation for bleeding, pulmonary complications, gastrointestinal complications, and return to the ICU. Women had significantly more intraoperative complications (RR = 2.09, 95% CI 1.35 3.23), length of stay (RR = 1.01, 95% CI 1.002-1.027), and low cardiac output conditions (RR = 1.10, 95% CI 1.01-1.20). Males developed more neurologic complications (RR = 0.85, 95% CI 0.75-0.95). CONCLUSIONS: Women undergoing coronary artery bypass graft surgery have more comorbidities at surgical presentation compared with men. Women have more unadjusted mortality, which is confounded by the difference in age and body surface area. But, even after multiple adjustments, female gender is a predictor of higher morbidity in patients undergoing coronary artery bypass graft. PMID- 12648696 TI - Surgical mortality in patients with infected aortic aneurysms. AB - BACKGROUND: Mortality in patients with infected aortic aneurysms remains high. A number of patient- and infection-specific risk factors for death have been proposed, but none is consistently predictive of poor outcomes. The purpose of this study was to examine the possible contribution of infection-related risk factors and the systemic inflammatory response syndrome (SIRS) to outcomes of patients with infected aortic aneurysms. STUDY DESIGN: Ten patients with infected aortic aneurysms presenting to our institution over a recent 6-year period were studied. Collected data included aneurysm location, culture results, preoperative indicators of SIRS, operative details, and outcomes. RESULTS: Common presenting symptoms included abdominal or back pain and fevers. Aneurysms involved the thoracoabdominal aorta in four patients, the suprarenal aorta in one, the juxtarenal aorta in one, and the infrarenal aorta in four. Seven patients met criteria for SIRS. Repairs included in situ replacement of the infected aneurysm using rifampin-soaked, gel-impregnated Dacron in four patients with thoracoabdominal aneurysms and using autogenous superficial femoral-popliteal vein in five patients with infrarenal aneurysms. Four patients died of sepsis, and six patients survived to discharge after a mean of 23 +/- 12 days in the hospital, followed by extensive rehabilitation. The combination of SIRS and suprarenal extension was present in all four patients who died. CONCLUSIONS: Although rare, infected aortic aneurysms are associated with marked morbidity and mortality. Sepsis is the leading cause of death. A combination of host- and infection-specific variables may be more predictive of outcomes than any single risk factor. Prolonged hospitalization and extended rehabilitation are frequently required in survivors, but longterm outlook is good after successful treatment. PMID- 12648698 TI - Selective cricopharyngeal myotomy for Zenker's diverticulum. PMID- 12648697 TI - Inguinofemoral dissection for carcinoma of the vulva: effect of modifications of extent and technique on morbidity and survival. AB - BACKGROUND: The aim of this study was to evaluate the impact of modifications of extent (medial inguinal and medial femoral lymphadenectomy, inguinal lymphadenectomy, inguinal and medial femoral lymphadenectomy, and inguinofemoral lymphadenectomy) and surgical technique of lymphadenectomy (including sartorius transposition, preservation of the fascia lata, and preservation of the saphenous vein) on morbidity, groin recurrence, and survival in patients with vulvar carcinoma. STUDY DESIGN: A retrospective review of 194 patients with primary squamous cell cancer of the vulva was conducted. Clinical, surgical, histopathologic, postoperative short- and longterm complications, and followup data were collected from patient records. RESULTS: Inguinal lymphadenectomy and medial inguinal and medial femoral lymphadenectomy produced about half fewer nodes than did other surgical procedures. On the other hand, number of lymph nodes removed did not differ notably between inguinofemoral lymphadenectomy and inguinal and medial femoral lymphadenectomy. Logistic regression showed that obesity was associated with increased risk of cellulitis. Age greater than 70, obesity, and extent of lymphadenectomy increased wound breakdown risk. Factors associated with leg edema persisting for more than 6 months were: extent of lymphadenectomy, sartorius transposition, and adjuvant irradiation of groin area. With a mean followup time of 38 months, neither groin recurrence rate nor disease specific survival markedly differed according to technique of lymphadenectomy. CONCLUSION: Techniques of lymphadenectomy with preservation of fascia lata and saphenous vein are associated with a decreased risk of postoperative morbidity without jeopardizing outcomes. PMID- 12648699 TI - What's new in plastic and maxillofacial surgery. PMID- 12648701 TI - Ethical considerations and barriers to research in surgical palliative care. PMID- 12648702 TI - Radiation arteritis. PMID- 12648703 TI - Congenital paraesophageal hiatal hernia. PMID- 12648704 TI - Tissue expansion-assisted closure of massive ventral hernias. PMID- 12648705 TI - The quick, no-twist, no-kink portal confluence reconstruction. PMID- 12648706 TI - History of surgery lecture. PMID- 12648708 TI - Postthyroidectomy hypocalcemia. PMID- 12648709 TI - Evidence-based research. PMID- 12648711 TI - [Randomised phase III trial of fotemustine versus fotemustine plus whole brain irradiation in cerebral metastases of melanoma]. AB - PURPOSE: The main objective of this prospective multicenter randomised phase III study was to compare a combined regimen of fotemustine plus whole brain irradiation versus fotemustine alone in terms of cerebral response and time to cerebral progression in patients with melanoma brain metastases. PATIENTS AND METHODS: Seventy-six patients (instead of the 106 planned patients; study was stopped after the interim analysis) were randomised receiving either fotemustine (arm A, n = 39) or fotemustine and whole brain irradiation (arm B, n = 37). Fotemustine was administered intravenously at 100 mg m(-2) on day 1, 8 and 15, followed by a 5-week rest period, then every 3 weeks in non-progressive patients. In arm B, a concomitant whole brain irradiation was performed at the total dose of 37.5 Gy (2.5 Gy/d(-1), days 1-5, 3 consecutive weeks). RESULTS: Although patients who received fotemustine alone had worse prognostic factors, there was no significant difference in brain response (arm A: 7.4%, B: 10.0%) or control rates (objective response plus stable disease) after seven weeks (arm A: 30%, B: 47%) and overall survival (arm A: 86d, B: 105d). However, there was a significant difference in favour of arm B for the time to brain progression (p = 0.028, Wilcoxon test). CONCLUSION: Fotemustine plus whole brain irradiation delayed the time to brain progression of melanoma cerebral metastases compared to fotemustine alone but without a significant improvement in terms of objective control or overall survival. PMID- 12648712 TI - [CT and MR image fusion using two different methods after prostate brachytherapy: impact on post-implant dosimetric assessment]. AB - PURPOSE: To study different methods of CT and MR images fusion in patient treated by brachytherapy for localized prostate cancer. To compare the results of the dosimetric study realized on CT slices and images fusion. MATERIALS AND METHODS: Fourteen cases of patients treated by I125 were retrospectively studied. The CT examinations were realized with contiguous section of 5mm thickness, and MR images were obtained with a surface coil with contiguous section of 3 mm thickness. For the images fusion process, only the T2 weighted MR sequence was used. Two processes of images fusion were realized for each patient, using as reference marks the bones of the pelvis and the implanted seeds. A quantitative and qualitative appreciation was made by the operators, for each patient and both methods of images fusion. The dosimetric study obtained by a dedicated software was realized on CT images and all types of images fusion. The usual dosimetric indexes (D90, V100 and V150) were compared for each type of image. RESULTS: The quantitative results given by the software of images fusion showed a superior accuracy to the one obtained by the pelvic bony reference marks. Conversely, qualitative and quantitative results obtained by the operators showed a better accuracy of the images fusion based on iodine seeds. For two patients out of three presenting a D90 inferior to 145 Gy on CT examination, the D90 was superior to this norm when the dosimetry was based on images fusion, whatever the method used. CONCLUSION: The images fusion method based on implanted seed matching seems to be more precise than the one using bony reference marks. The dosimetric study realized on images fusion could allow to rectify possible errors, mainly due to difficulties in surrounding prostate contour delimitation on CT images. PMID- 12648713 TI - [Primary intramedullary ependymomas: retrospective study of 16 cases]. AB - PURPOSE: To review the radiotherapy department experience in treating primary spinal cord ependymomas (PSCE), analyse prognostic factors and provide treatment recommendations regarding literature review. MATERIALS AND METHODS: Sixteen patients with PSCE received postoperative radiotherapy between 1972 and 1997. There were 10 male and 6 female patients with a mean age of 34 years (range 2 63). Surgery was gross total resection in 2 cases, subtotal resection in 9, biopsy in 4 and of unknown type in one patient. All patients were treated with radiotherapy to the craniospinal axis (4 cases), spinal cord (1 case) and to the site of primary tumour in 11 cases. RESULTS: Five year-survival rate was 73%. Two patients had recurrent tumours within the primary site. Gender, extent of surgery and treatment field extent were not prognostic factors. Histologic type was the only variable predictive of outcome. Patients with myxopapillary type had a 5 year survival rate of 100% compared with 47% for those with other histology types. CONCLUSION: We conclude that aggressive surgery is not necessary in the management of PSCE, localised field radiotherapy is associated with favourable outcome, and tumour grade is an important prognostic factor. PMID- 12648714 TI - [Brachytherapy in carcinomas of anal canal and rectum: techniques and results]. AB - Conservative treatment of small anal canal and low rectal cancers is a valid alternative to radical surgery through combinated strategies including external beam and/or intracavitary irradiation, concomitant chemotherapy and 192 iridium implantation as a booster dose. Feasibility, tolerance and results are effective. Local control of the disease and the quality of the anal functions are good. The treatment requires great care in patient selection and follow-up. PMID- 12648715 TI - [Endobronchial brachytherapy: technique and indications]. AB - The endobronchial brachytherapy procedure involves the insertion of an afterloading catheter into bronchus in close proximity to an endoluminal lesion, and to perform limited irradiation sparing as much as possible normal health tissues. The catheter is inserted during a classical flexible bronchoscopy. Three types of indications have been discussed: (i) palliative treatment of lung carcinoma, with or without laser desobstruction: an improvement in respiratory symptoms was observed in 60 to 80% of the cases; (ii) curative treatment for localised endobronchial carcinomas, in previously irradiated patients, or in case of contraindication of surgery or external beam irradiation; local control rate range from 60 to 70% at 2 years; (iii) combination of external irradiation and brachytherapy in the first line treatment of lung cancers. Two randomised trials did not show any improvement in survival for this approach; however, they have included advanced diseases. In the opposite, this association seems very effective for early stage lung carcinomas. Two major complications were regularly reported, haemoptysis and radiation bronchitis. Predictive factors for these toxicities are actually better known: haemoptysis could be due to a progressive disease more often than to brachytherapy itself; technical factors (dose, volume, fractionation), however, could explain a number of radiation bronchitis, and their incidence could decrease in the future. PMID- 12648716 TI - [Brachytherapy in cervix cancers: development of techniques and concepts]. AB - Brachytherapy plays an important role in the treatment of patients with cervical carcinoma. Technical modalities have evolved during the last years and have benefited from imaging modalities development, specially MRI. Imaging modalities contribute to a better knowledge of tumoral extension and critical organs. Ultrasound during brachytherapy has led to the almost complete eradication of uterine perforation. In the future, a more systematic use of systems allowing optimization may induce a better dose distribution in the tumor as well as in the critical organs. Recent data provided information in favor of a better analysis in the relative role of dose-rate, total dose and treated volume and their influence on the local control and complication incidence. Concomitant radiochemotherapy represents a standard in the treatment of patients with tumoral size exceeding 4 cm. Some questions still remain: is concomitant chemotherapy of benefit during brachytherapy? Is there any place for complementary surgery, specially in patients with complete response after external irradiation with concomitant chemotherapy and brachytherapy? In order to answer the former question, a phase III randomized trial is going to start, with the Federation Nationale des Centres de Lutte Contre le Cancer as a promoter. PMID- 12648717 TI - [Brachytherapy for coronary restenosis: state of art in 2003]. AB - Based on therapeutic approach for benign diseases, vascular brachytherapy decreases smooth vascular muscle cells proliferation and multiplication which lead to the formation of the neo-intima. The radioactive positive action affects arterial recoil due to post angioplasty vessel injury. Randomised studies has shown good angiographic results up to 6 months of follow-up, with 50% in-stent restenosis rate decrease and on the analysed segment as well. Decrease on Mace and TLR show statistically significance. Results don't correlate with emitter and beta emitters had been introduced in France recently. Vascular brachytherapy is actually indicated for in-stent restenosis, there is no evidence to perform this treatment for de novo lesion. Geographic miss, source centering, late thrombosis and pullback procedure may interfere with treatment quality. IVUS allows best target volume determination to a higher quality level. Internationals guidelines such as Eva-Gec-Estro recommendations could increase treatment safety and enable development of an optimal technique. PMID- 12648719 TI - [Brachytherapy in soft tissue sarcomas]. AB - Soft tissue sarcomas are rare tumors with a high risk or local recurrence and a risk of distant metastases. Standard treatment advocated is the combination of conservative resection and external radiotherapy. Brachytherapy is an integrated part of the multidisciplinary treatment. Brachytherapy can increase local control with good functional results. Primary exclusive brachytherapy has been used and is effective and safe in high grade sarcomas (randomized trial of MSKCC). Brachytherapy seems to be important as part of the treatment of central localization (shoulder, groin) and sarcomas with positive resection margins, but its relation with external radiotherapy has to be defined. Brachytherapy used with special guidelines allows to obtain an improved local control with an acceptable level of complication. PMID- 12648718 TI - [Brachytherapy in head and neck cancers]. AB - Experience accumulated over several decades with radiation of Head and Neck tumours by irradiation has demonstrated the need for a high tumour dose to achieve local control. With external beam irradiation alone, it is difficult to spare adjacent normal tissues, resulting in undesirable late effects on the salivary glands, mandible, and muscles of mastication. Interstitial implantation is ideally suited to deliver a high dose limited to the volume of the primary tumor, thus minimizing sequels. A large experience has been accumulated with low dose rate (LDR) brachytherapy in treatment of carcinoma of oral cavity, oropharynx, and nasopharynx. Recent analysis of large clinical series provided data indicating that modalities of low dose rate brachytherapy should be optimized in treating these tumors for increasing therapeutic ratio. Low dose rate brachytherapy is now challenged by high dose rate (HDR) brachytherapy and pulsed dose rate (PDR) brachytherapy. Preliminary results obtained with these two last modalities are discussed regarding to those of low dose rate brachytherapy. PMID- 12648720 TI - Deciphering the molecular basis of multidrug recognition: crystal structures of the Staphylococcus aureus multidrug binding transcription regulator QacR. AB - Multidrug transporters and their transcriptional regulators are key components of bacterial multidrug resistance (MDR). How these multidrug binding proteins can recognize such chemically disparate compounds represents a fascinating question from a structural standpoint and an important question in future drug development efforts. The Staphylococcus aureus multidrug binding regulator, QacR, is soluble and recognizes an especially wide range of structurally dissimilar compounds, properties making it an ideal model system for deciphering the molecular basis of multidrug recognition. Recent structures of QacR have afforded the first view of any MDR protein bound to multiple drugs, revealing key structural features of multidrug recognition, including a multisite binding pocket. PMID- 12648721 TI - Genetic polymorphism of cyanobacteria under permanent natural stress: a lesson from the "Evolution Canyons". AB - Cyanobacteria have high adaptive potential and occur in the most extreme habitats. The available literature data indicate that the versatility of cyanobacteria is related to their higher polymorphism under stress. The studies of a filamentous cyanobacterium, Nostoc linckia, from the ecological microsite models known as "Evolution Canyons" showed that, among the evolutionary forces maintaining the higher polymorphism and genome diversity under permanent natural stress, the various types of natural selection play a key role. PMID- 12648723 TI - Burkholderia cenocepacia sp. nov.--a new twist to an old story. AB - DNA-DNA hybridisation experiments between isolates representing Burkholderia cepacia genomovar III recA lineages IIIA and IIIB reinforced the classification of both phylogenetic subgroups as a single genospecies, distinct from B. cepacia (genomovar I). A formal classification of B. cepacia genomovar III encompassing the recA lineages IIIA and IIIB, and the new recA lineages IIIC and IIID, as B. cenocepacia sp. nov., with LMG 16656 as the type strain, is proposed. PMID- 12648722 TI - Pfiesteria piscicida, P. shumwayae, and other Pfiesteria-like dinoflagellates. AB - Pfiesteria piscicida and Pfiesteria shumwayae are estuarine dinoflagellates thought to be responsible for massive fish deaths and associated human illnesses in the southeastern United States. These dinoflagellates are described as having a complex life cycle involving flagellated zoospores, cysts, and amoeboid stages. Although no Pfiesteria toxin has been identified, certain strains of these dinoflagellates are thought to produce a water-soluble toxin that can kill fish and cause human illness. Recent reports show no evidence for amoeboid stages and indicate that a much more simplified life cycle exists. In addition, researchers have shown that P. shumwayae only kills fish through direct contact that does not necessarily involve the production of one or more toxins. This review summarizes these and other recent findings with an emphasis on establishing basic facts regarding the toxicity and life history of Pfiesteria dinoflagellates. PMID- 12648724 TI - Detection and characterization of verotoxigenic Escherichia coli by a VTEC/EHEC multiplex PCR in porcine faeces and pig carcass swabs. AB - To determine whether pigs and pig carcasses are a reservoir of verotoxigenic producing Escherichia coli, a sensitive and rapid isolation method was developed. A VTEC/EHEC multiplex PCR method was developed to test all samples taken at the farm (n=289) and at the slaughterhouse (n=233). From the 137 positive VTEC/EHEC multiplex PCR farm samples, 54 strains were isolated that carried one virulence gene. Of these, thirty-one strains carried the vt2e variant of the vt2 gene, which causes oedemic disease in young pigs, four strains contained the hlyA gene and 19 the eaeA gene. All strains isolated were further identified with an enterotoxigenic E. coli (ETEC) multiplex PCR developed to detect the LTI, STIa, STII and vt2 genes in a single reaction. One-third of the vt2e-positive strains also contained the STIa gene. A small fraction of the 132 carcasses tested were found positive for VTEC or the E. coli O157 serotype. We conclude that none of the isolated strains carried a combination of virulence genes indicative of potential human pathogenic characteristics. PMID- 12648725 TI - Fluctuation of bacteria isolated from elm tissues during different seasons and from different plant organs. AB - In this work we isolated a culturable endophytic aerobic heterotrophic bacterial community from the stem and root tissues of elm trees (Ulmus spp.) and analyzed its fluctuations. A total of 724 bacterial isolates were collected at different times (April, June, September and December) from two elm trees, one infected with Elm Yellows phytoplasmas, and one which was healthy-looking. The isolates were grouped into 82 haplotypes, identified by means of amplified ribosomal DNA restriction analysis (ARDRA) using the restriction enzyme AluI, suggesting that the genetic diversity of the bacterial community was very high. The taxonomic position of the isolates belonging to the twelve main haplotypes, representing more than 72% of the total population, was determined by 16S rDNA sequencing. The main genera were Bacillus, Curtobacterium, Pseudomonas, Stenotrophomonas, Sphingomonas, Enterobacter, and Staphylococcus. The fluctuations in the bacterial community, determined by different parameters (seasonal changes, plant organ, presence of phytoplasmas) were studied, revealing that they were influenced both by variations in temperature (warm or cold according to the season) and by the organ examined (roots or stems). The role of the phytopathogenic status in these fluctuations was also discussed. PMID- 12648726 TI - The effect of acid treatment on survival and protein expression of a laboratory K 12 strain Escherichia coli. AB - Pre-exposure of log phase enteric bacteria to nonlethal acidic pH induces phenotypic changes that protect the organisms against subsequent lethal acidity. Studies have revealed that when Salmonella typhimurium is grown in minimal medium at pH 5.5 and 4.3 the organism develops a biphasic acid tolerance. This two-stage response has not been reported at present in Escherichia coli; rather it is thought that when this organism is grown in rich medium there is a single stress response throughout the pH range of 4 to 6. We believe that the evidence for such a report is lacking; therefore, in this study the acid response of log phase E. coli was examined in rich medium (LB). The pH 3.0 acid survival assays of a laboratory strain of E. coli K-12 MG1655, after cultures had been exposed to LB acidified to pH 5.5 or pH 4.3 indicate that like S. typhimurium, E. coli shows both an acid tolerance and an acid-shock response to pH 5.5 and 4.3 exposure, respectively. It was consistently found, however, that longer pre-exposure (60 min rather than 15 min) at either pH afforded better protection against the lethal pH 3.0 challenge. Analysis of polypeptide induction at pH 5.5 and 4.3 by two-dimensional gel electrophoresis clearly shows different profiles. Together the results show that in E. coli, pre-treatment between pH 4 and 6 does not result in a flat protective response. PMID- 12648727 TI - Identification and characterization of the new gene rhtA involved in threonine and homoserine efflux in Escherichia coli. AB - The rhtA gene known as the ybiF ORF in the genome of Escherichia coli was identified as a new gene involved in threonine and homoserine efflux. This gene encodes a highly hydrophobic membrane protein that contains 10 predicted transmembrane segments. The rhtA23 mutation, which is an A-for-G substitution at position -1 in relation to the ATG start codon, increases the expression level of the rhtA gene. The overexpression of rhtA gene results in resistance to inhibitory concentrations of homoserine, threonine and a variety of other amino acids and amino acid analogues, reduced threonine and homoserine accumulation in resistant cells and increased production of threonine, homoserine, lysine and proline by the respective producing strains. The RhtA protein belongs to a vast family of transporters. The genome of E. coli contains at least 10 paralogues of RhtA. Phylogenetic analysis indicates that a common ancestor of living organisms contained several RhtA homologues. PMID- 12648728 TI - Characterization of the genes rdxA and frxA involved in metronidazole resistance in Helicobacter pylori. AB - Metronidazole (Mtz) resistance in Helicobacter pylori has been found to be associated with mutations in rdxA, a gene encoding an oxygen-insensitive NADPH nitroreductase, and enhanced by mutations in frxA, a gene encoding a NAD(P)H flavin oxidoreductase. The roles of these two genes in Mtz resistance in H. pylori were examined in this study. The rdxA and frxA genes were sequenced in nine pairs of strains isolated from biopsies obtained from patients before and after failed eradication treatments which included Mtz and resulted in the appearance of resistant strains. Metronidazole resistance could be explained in seven of these pairs of strains by mutations in rdxA and frxA. However, in one pair of strains, rdxA was identical in the susceptible and resistant strains, and only changes in frxA were observed; and in another pair, neither rdxA nor frxA were different in the susceptible and resistant strains. Sequencing of the upstream region of frxA and of the recA gene in the latter pair of strains did not reveal any mutations. To establish whether mutations in frxA alone could be involved in Mtz resistance, a resistant Escherichia coli strain transformed with the frxA of a Mtz susceptible H. pylori strain was rendered susceptible, and transformation with a mutated H. pylori frxA gene under the same conditions did not change the resistant E. coli phenotype. The results suggested that a Mtz resistance phenotype may arise in H. pylori without mutations in rdxA or frxA, or with mutations only in frxA. PMID- 12648729 TI - Rapid detection and identification of Brachyspira aalborgi from rectal biopsies and faeces of a patient. AB - This study reports for the first time the detection of Brachyspira aalborgi in faeces and rectal biopsies of a female suffering for 3-4 months of abdominal pain with long-standing mucosal diarrhoea, rectal bleeding and suspected carcinoma of the rectum. After pre-treatment of samples (faeces and biopsies) with a liquid medium (trypticase soy broth-TSB) containing foetal calf serum (FCS, 10%) and spectinomycin and rifampicin (TSB-SR) the first detection of B. aalborgi isolate HBS1 was observed after 48 h in the primary plates of selective blood agar modified medium (BAM) containing spectinomycin and rifampicin (BAM-SR), where growth zones were signalled by a small weakly beta-haemolytic halo. Attempts to subculture spirochaetes in agar media failed. The new HBS1 isolate was only propagated in TSB broth and at electron microscopy it showed 4 endoflagella inserted at each tapered end. The phenotypic characterization of HBS1 demonstrated absence of hippurate hydrolysis, indole production, alpha galactosidase, alpha- and beta-glucosidase activities in accordance with the B. aalborgi type strain. Rapid identification of B. aalborgi isolate HBS1 was performed directly from faeces and rectal biopsies and subsequently from pure cultures by a genetic method based on 16S DNA restriction fragment length polymorphism (RFLP)-polymerase chain reaction (PCR). The sequence of 16S DNA amplicon of the isolate HBS1 was found 99.2% corresponding to that of the B. aalborgi type strain. Our results encourage further investigations for the development of a suitable selective agar medium for the isolating and cultivating B. aalborgi from human specimens. PMID- 12648730 TI - Increased phospholipase A2 activity in schizophrenia with absent response to niacin. AB - An absent response to the niacin skin test has been reported to occur in about 80% of schizophrenic patients, as compared to 20% of healthy individuals. Niacin provokes redness in skin caused by a capillary vasodilatation mediated by prostaglandins. The metabolism of prostaglandins is regulated by the enzyme phospholipase A2 (PLA2). Several studies have reported increased PLA2 activity in schizophrenia. In this study we investigated the relationship between niacin response and PLA2 activity in 38 drug-free schizophrenic patients and in 28 healthy controls. Twenty-two of these patients were reevaluated after 8 weeks under treatment with new generation antipsychotic drugs. Niacin response was absent in 23% of the schizophrenic patients and in 14% in controls (n.s.). PLA2 activity was higher in schizophrenics than in controls (344+/-115 vs. 290+/-71 pmol/ml/min; p=0.03). Patients with absent response to niacin had the highest PLA2 activity as compared to those with positive response (426+/-155 vs. 319+/ 111; p=0.02). After 8 weeks on antipsychotic treatment, PLA2 activity was reduced (355+/-115 before, 267+/-39 after, p=0.001) and 4 out of 13 patients with absent response to niacin converted to positive. The reduction of PLA2 activity in these patients was higher than in patients who remained with absent response (36% vs. 23%). Our data support the findings that absent response to niacin is more frequent in schizophrenic than in healthy individuals although the magnitude of the difference was smaller than that reported in the literature. The relationship between absent response to niacin in schizophrenia and increased PLA2 activity suggests further that the skin test may be useful to easily identify a subgroup of patients with a disordered phospholipid metabolism. PMID- 12648731 TI - The PRIME North America randomized double-blind clinical trial of olanzapine versus placebo in patients at risk of being prodromally symptomatic for psychosis. I. Study rationale and design. AB - The first double-blind placebo-controlled clinical trial of an atypical neuroleptic medication is being conducted in symptomatic treatment-seeking patients meeting new diagnostic criteria for a putative prodromal syndrome. This identifies them as being at high risk for developing psychosis in the near future. The study aims include prevention of psychosis onset and disability, as well as palliation of ongoing symptomatology. This report presents the study rationale and design. Recent studies will be reviewed that have advanced our knowledge about the early course of schizophrenia and our ability to predict onset prospectively, advances that have rendered prodromal intervention research feasible and ethical. The study design has many novel features. It tests for prevention versus delay in psychosis onset, as well as for efficacy and safety in a newly defined clinical population. This has required the development of innovative clinical research assessment instruments and a new operational definition of psychosis onset. The integration of these novel elements into an otherwise typical clinical trial design is detailed. The companion report will address sample recruitment and the clinical phenomenology at baseline of this putative "prodromal" entity. PMID- 12648732 TI - The PRIME North America randomized double-blind clinical trial of olanzapine versus placebo in patients at risk of being prodromally symptomatic for psychosis. II. Baseline characteristics of the "prodromal" sample. AB - The first double-blind placebo-controlled clinical trial of an atypical neuroleptic medication is being conducted in symptomatic treatment-seeking patients meeting new diagnostic criteria for a putative prodromal syndrome. This identifies them as being at high risk for developing psychosis in the near future. The study aims include prevention of psychosis onset and disability, as well as palliation of ongoing symptomatology. The purpose of this report is to describe the study's "prodromally symptomatic" sample at baseline, i.e., at intake immediately prior to randomization and prior to receiving study medication. Sixty treatment-seeking patients meeting prodromal inclusion criteria were recruited across four sites: New Haven, CT (n=39), Toronto, Ontario (n=9), Calgary, Alberta (n=6), and Chapel Hill, NC (n=6). The sample was young (median age 16), largely male (65%), and came from families with high titers of serious mental illness (44%). Most patients (93%) met criteria for the Attenuated Positive Symptom (APS) prodromal syndrome and presented with significant but nonpsychotic suspiciousness, perceptual aberrations, unusual thought content, and conceptual disorganization. They presented with minimal to mild affective symptoms and substance use/abuse, but they were quite functionally compromised (mean Global Assessment of Functioning (GAF) score=42). The prodromal sample was compared with other clinical-trial samples of adolescent depression, adolescent mania, and first episode schizophrenia. Prodromal patients proved not to be depressed or manic. They were less severely ill than untreated first episode schizophrenia but more severely ill than treated first episode schizophrenia. While not psychotically disabled, these patients nevertheless present with a clinical syndrome. Subsequent reports will detail the effects of drug versus placebo on prodromal symptoms, neuropsychological profile, and the rate of conversion to psychosis. PMID- 12648734 TI - TNFB polymorphism may be associated with schizophrenia in the Korean population. AB - This study was conducted to test the association between the tumor necrosis factor (TNF)-beta gene (B) polymorphism and schizophrenia in the Korean population. 127 patients with schizophrenia according to the DSM-IV criteria and 202 healthy controls were enrolled in this study. Patients and controls were biologically unrelated age and sex-matched native Koreans. Genotyping for the TNFB polymorphism was performed by polymerase chain reaction-restriction fragment length polymorphism. Genotype and allele distributions of the TNFB polymorphism in patients with schizophrenia were significantly different from those of the controls. Subjects with the TNFB*2 allele had an increased risk for schizophrenia (Odds Ratio=1.76, 95% CI=1.27-2.45). The present study suggests that the TNFB polymorphism may confer susceptibility to schizophrenia in the Korean population. PMID- 12648733 TI - Association between a promoter variant in the monoamine oxidase A gene and schizophrenia. AB - Monoaminergic transmission has been implicated in the pathophysiology of schizophrenia. We investigated a putative functional promoter polymorphism in the monoamine oxidase A (MAOA) gene in schizophrenic patients (n=133) and control subjects (n=377). In men, there was an association between the less efficiently transcribed alleles and schizophrenia (chi(2)=4.01, df=1, p<0.05). In women, no significant differences were found. The present results support the involvement of the MAOA gene in men with schizophrenia in the investigated Swedish population but should be interpreted with caution until replicated. PMID- 12648735 TI - The factor structure for the Positive and Negative Syndrome Scale (PANSS) in recent-onset psychosis. AB - The Positive and Negative Syndrome Scale (PANSS) is a widely used instrument for measuring severe psychopathology in adult patients with schizophrenia. Data, primarily on chronic patients, have been used to define factors for the PANSS. The present study examines the PANSS factor structure in a large sample of subjects with recent-onset schizophrenia, schizophreniform disorder and schizoaffective disorder who had been exposed to very limited antipsychotic medication. Equamax factor analysis was conducted on PANSS baseline assessments from a multicenter, 11 country drug trial that enrolled 535 patients. The forced five-factor solution essentially corresponds to the factors most frequently described previously, namely negative, positive, disorganized (or cognitive), excited and anxiety/depression. In the exploratory analysis, a seven-factor solution was obtained, with depression and anxiety symptoms separating and a motor component emerging. The results of this study partially support the use of a five-factor model for the PANSS, but suggest that scales for catatonia, depressive and anxiety syndromes should be included in future studies. PMID- 12648736 TI - Anhedonia in schizophrenia: a distinct familial subtype? AB - Failures to replicate results in psychiatric genetics might be due to our inability to define the heritable phenotype. Instead of relying entirely on classical nosographical approaches, the use of a candidate symptom approach to identify more homogeneous forms of diseases among affected subjects and subclinical traits among first-degree relatives may increase genetic validity. Anhedonia may be a marker for subjects at risk of schizophrenia or schizophrenia spectrum disorders. We compared the familiality of anhedonia characterized by a high level of physical anhedonia (score above 23) in a sample of schizophrenic probands (N=80) and their relatives (N=78), with that in bipolar patients (N=109), their relatives (N=33) and normal controls (N=94). We identified a subform of schizophrenia characterized by highly anhedonic schizophrenic probands with a three-fold higher familial risk of schizophrenia and schizophrenic spectrum disorders. We also found that their first-degree relatives had a high level of anhedonia. An intrafamilial correlation analysis confirmed the familial nature of anhedonia. Our data suggest that anhedonia is a candidate symptom for schizophrenia. Refining phenotype definition by studying subgroups of anhedonic and non-anhedonic probands with relevant candidate genes might be fruitful. PMID- 12648737 TI - Minor physical anomalies in schizophrenia: is age a confounding factor? AB - Minor physical anomalies (MPAs) occur more frequently in a range of developmental disorders. They are also more frequent in schizophrenia supporting a neurodevelopmental aetiology of the illness. Contemporary MPA scales are yet to be validated for the effects of age. It is hypothesised that the effects of ageing may be confounding when these scales are applied to an elderly population. The distribution of MPAs in a normal elderly population was compared with younger subjects. Fifty subjects over the age of 60, and 50 subjects below the age of 60, with no known major mental illness, were evaluated. MPAs were assessed using a modified Lane scale [Psychol. Med. 27 (1997) 1155]. Elderly subjects showed an excess of absent trichions due to alopecia (p=0.004) short and broad palates (p=0.01) and greater ear protrusion (p=0.004) relative to young subjects. The differences reported are probably due to hair loss, edentulousness and growth of the auriculocephalic angle with age. These findings question the validity of studies of MPAs in schizophrenia that do not control for age. PMID- 12648738 TI - Insight in schizophrenia: a meta-analysis. AB - There has been an increase in the study of insight in schizophrenia in the last 20 years. Insight is operationally defined according to five dimensions which include: the patient's awareness of mental disorder, awareness of the social consequences of disorder, awareness of the need for treatment, awareness of symptoms and attribution of symptoms to disorder. Despite the development of psychometrically sound measurement tools, the results from previous studies have been inconclusive regarding the nature of the relationship between insight and symptomatology. A meta-analysis of 40 published English-language studies was conducted to determine the magnitude and direction of the relationship, or effect size, between insight and symptom domains in schizophrenia and to determine moderator variables that were associated with the variations in effect sizes across studies. Results indicated that there was a small negative relationship between insight and global, positive and negative symptoms. There was also a small positive relationship between insight and depressive symptoms in schizophrenia. Acute patient status and mean age of onset of the disorder moderated the relationship between insight and symptom clusters. The possible reasons for the effect sizes being modest, the examination of the role of moderator variables and directions for future research are provided. PMID- 12648739 TI - Anxiety and substance use comorbidity among inpatients with schizophrenia. AB - OBJECTIVE: To determine the association between lifetime anxiety symptoms and anxiety disorders and substance use disorders among patients with schizophrenia. METHOD: Participants were 184 inpatients with schizophrenia at the Schizophrenia Research Unit (SRU) at the New York State Psychiatric Institute (NYSPI). Multivariate logistic regression analyses were used to determine the relationship between specific anxiety symptoms and anxiety disorders and substance use disorders among inpatients with schizophrenia. RESULTS: Anxiety symptoms and anxiety disorders were prevalent among 31.5% of the sample. Panic attacks were associated with a significantly increased odds (OR=7.4 (1.2, 47.1)) of comorbid alcohol or substance use disorders (lifetime). This association was specific to panic attacks and persisted after adjusting for differences in sociodemographic characteristics and comorbid anxiety symptoms and anxiety disorders. CONCLUSIONS: These findings are consistent with and extend previous data by providing evidence of an association between panic attacks and increased likelihood of substance use disorders among inpatients with schizophrenia. Future studies that determine the nature of this relationship, the sequence of symptom onsets, and examine whether treatment of anxiety can influence the onset or outcome associated with substance use are needed. PMID- 12648740 TI - A confirmatory factor analytic evaluation of the pentagonal PANSS model. AB - The positive and negative syndrome scale (PANSS) is widely used in psychiatric research. Reflecting this common use, considerable attention has been applied to the psychometric properties of this instrument. However, despite the publication of numerous studies and analyses, it remains uncertain how best data from the PANSS should be analysed to best model the symptoms of schizophrenia. A resolution to these concerns seemed to be offered following the publication in 1997 of a large multisite factor analysis that produced the 'pentagonal model', which has subsequently been included in the 2000 revision of the PANSS user manual. However, to date, an independent confirmatory analysis of this model has not yet been published. The aim of this study was to test this model in a new independent sample with confirmatory factor analysis (CFA). Independent confirmation of the fit of the model is required to ensure that its implementation is informed by confirmation of its psychometric properties. CFA was performed in a sample of 347 subjects with schizophrenia. The analysis found that the model had inadequate goodness of fit. The use of the pentagonal model has similar difficulties as earlier models and more research is required to ascertain the optimal method for measuring symptom dimensions in research and clinical settings. PMID- 12648741 TI - Increasing age does not decrease risk of schizophrenia up to age 40. AB - It is well established that at the population level the age-specific incidence of schizophrenia is highest in the early 20s and decreases with age. We present new findings showing that this is not the case when we assume that only part of the population is susceptible. The study population consisted of birth cohorts born in Finland from 1950 to 1968. These were identified from the Population Register Center, which provided the data as multidimensional tables with sex, and year, month, and place of birth as marginals. Each cohort was followed-up until 1991, up to age 40. Data analyses were based on a compound Poisson statistical model, which assumed that only part of the population is susceptible to develop schizophrenia. The method detects the proportion of susceptibles in population level, but does not identify susceptible individuals. The model was able to reveal the basic hazard distribution, which showed increasing risk of developing schizophrenia with higher age at the individual level among the susceptible part of the population. The increase in risk was more rapid among males, but the susceptible part of the population was of similar size among both sexes. The proportion of susceptibles seemed to be lower in younger cohorts. We conclude that when an individual belongs to the susceptible part of the population, his/her individual risk of developing schizophrenia increases with age, at least up to age 40. This increase is more rapid among males. The reasons behind the lower proportion of susceptibles in younger birth cohorts, as well reasons for differences in risk between sexes remained unclear and need more research. PMID- 12648742 TI - Dopamine D4 receptor gene (DRD4) variants and schizophrenia: meta-analyses. PMID- 12648743 TI - EEG brain mapping in schizophrenic patients and healthy subjects during facial emotion recognition. PMID- 12648744 TI - Role of the dorsomedial hypothalamus in thermogenesis and tachycardia caused by microinjection of prostaglandin E2 into the preoptic area in anesthetized rats. AB - Prostaglandin E2 (PGE2) acts in the preoptic area (POA) of the mammalian hypothalamus to increase body temperature and heart rate. Chemical stimulation of the dorsomedial hypothalamus (DMH), a region richly innervated by neurons in the POA, evokes sympathetically-mediated increases in heart rate and body temperature. We tested the hypothesis that neurons in the DMH mediate hyperthermia and tachycardia resulting from the action of PGE2 in the POA. Microinjection of PGE2 150 pmol/15 nl into the POA in urethane-anesthetized rats caused increases in body temperature and heart rate that were sharply reversed after injection of muscimol 80 pmol/100 nl into the DMH but not after similar injection of saline vehicle. Therefore, thermogenic and tachycardic actions of PGE2 in the POA are at least in part a consequence of neuronal activity in the region of the DMH. PMID- 12648745 TI - Abnormal response to glutamate of T lymphocytes from multiple sclerosis patients. AB - Multiple sclerosis (MS) is an immune-mediate, inflammatory and demyelinating disease of the central nervous system (CNS). Since glutamate (Glu) is a modulator of T lymphocyte function and Glu excitotoxicity has been proposed as one of the mechanisms of the demyelination, we studied the responses of T lymphocytes from normal controls (NC), MS or other non-inflammatory neurological disease (ONND) patients to Glu, by measuring phytohemagglutinin-induced intracellular Ca(2+) ([Ca(2+)](i)) rise (Fura-2 method) and cell proliferation (3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyl tetrazolium bromide colorimetric assay). No differences in the Glu (1 microM)-induced potentiation of the [Ca(2+)](i) rise were measured in T lymphocytes from all groups of subjects, while a significant decrease in the Glu (1 mM)-induced inhibition of cell proliferation was observed in T lymphocytes from MS patients. These data demonstrate that MS T lymphocytes abnormally respond to Glu. PMID- 12648746 TI - Neuropeptide Y modulates a G protein-coupled inwardly rectifying potassium current in the mouse hippocampus. AB - Neuropeptide Y (NPY) is an abundant brain peptide with endogenous antiepileptic activity. Here we examined the role played by Y1 receptors (Y1R) in the mouse hippocampus. Using whole-cell patch-clamp recordings, we show that hilar neurons in acute mouse hippocampal slices exhibit a G-protein coupled inwardly rectifying potassium (GIRK) current that is significantly enhanced during exogenous NPY application. NPY-mediated enhancement of GIRK current was observed on 47% of putative interneurons and was mimicked by application of Y1R specific agonist (Leu(31)Pro(34) NPY). Immunostaining revealed the presence of Y1R on cell somas of hilar NPY-containing interneurons. Thus, our results suggest that Y1R on hilar interneurons may act as a peptide autoreceptor. PMID- 12648747 TI - Discrimination of Exner's area and the frontal eye field in humans--functional magnetic resonance imaging during language and saccade tasks. AB - In the left frontal lobe, Exner's area (EXA), which is responsible for writing and reading, is located close to the frontal eye field (FEF), which is responsible for eye movements. To discriminate EXA from FEF anatomically and functionally, functional magnetic resonance imaging was conducted in 12 healthy volunteers. The saccadic eye movement experiment activated a region defined as the FEF, whereas three language experiments that included translation between grapheme and phoneme activated another region defined as EXA. EXA was found to be located only 1.5 cm apart from the FEF in the Talairach brain template. By conducting the saccade and language experiments in the same individuals, this study was able to successfully separate EXA from FEF. PMID- 12648748 TI - Subjective sleepiness correlates negatively with global alpha (8-12 Hz) and positively with central frontal theta (4-8 Hz) frequencies in the human resting awake electroencephalogram. AB - Subjective sleepiness is part of the system controlling the decision to go to sleep in humans. Extended periods of waking lead to increased sleepiness, as well as to changes in cortical electroencephalogram (EEG) during waking. We investigated the association of sleepiness and awake EEG spectra during 40 h of wakefulness using multi-electrode EEG recordings for full coverage of the scalp. We found: (1). strong negative correlations of alpha (8-12 Hz) power with subjective sleepiness at all scalp locations, suggesting a negative association between sleepiness and general cortical activation; and (2). positive correlations of theta (4-8 Hz) power with subjective sleepiness with a focus on frontal locations, suggesting additional location specific associations between sleepiness and cortical activation. These findings support the notion that sleepiness is directly represented in the awake EEG. PMID- 12648749 TI - Selective attention deficits during human pregnancy. AB - Using a longitudinal design we provide evidence that selective attention - a key component of cognition concerned with selection and preparation - is compromised during pregnancy. Selective attention was operationalized by means of the finger precuing technique, which selectively prepares two of four finger responses. The precuing benefit was taken as a measure of selective attention. Pregnant women showed a significant smaller precuing benefit at week 36 of pregnancy than did the control women, indicating loss of selective attention. Thirty-two weeks after childbirth this performance decrement had vanished, reflecting a functional recovery. PMID- 12648750 TI - Current source density distribution of sleep spindles in humans as found by synthetic aperture magnetometry. AB - Previous magnetoencephalography (MEG) studies of sleep spindles have identified a complex and widespread distribution in parietal and frontal cortices by fitting a dipolar model to the data. In this study, we used a whole-head 151-channel MEG system with simultaneous electroencephalography (EEG) recording in eight normal subjects. All subjects fell asleep during stage 2 and 3; EEG spindles have been observed in all cases. The current source density distribution of sleep spindles in the 10-15 Hz frequency band was localized by means of synthetic aperture magnetometry, and statistically tested using a permutation analysis. Source locations of the sleep spindles were found primarily in the frontal cortex, including dorsolateral and medial prefrontal areas, as well as the parietal cortex, including the vicinity of the primary sensorimotor areas. These results suggest that sleep spindles are similar in frequency content and spatial location to mu rhythm, and that these two oscillatory activities might also have a common neural basis and physiological meaning. PMID- 12648751 TI - An association study of the codon 72 polymorphism in the pro-apoptotic gene p53 and Alzheimer's disease. AB - Recent studies have demonstrated that p53-associated apoptosis is involved in the pathogenesis of Alzheimer's disease (AD). We performed a case-control association study between sporadic AD and the common proline/arginine polymorphism at codon 72 in the pro-apoptotic gene p53, in 109 sporadic AD patients and in 111 controls. This polymorphism has been intensively investigated for association with cancer, but so far not with AD and neurodegeneration. We found no association between this locus and the risk for AD. No association was detected also for the age at disease onset and for disease progression, and no interactive effect was found with apolipoprotein E e4. These findings show no evidence for an association between p53 codon 72 polymorphism and AD in our population. PMID- 12648752 TI - Reduced expression of calsenilin/DREAM/KChIP3 in the brains of kainic acid induced seizure and epilepsy patients. AB - Calsenilin is a neuronal calcium binding protein that may function in calcium signaling and cell death. Kainic acid, an analog of the excitatory amino acid L glutamate, produced excitotoxic cell death and induced the pathophysiology of status epilepticus. The expression of calsenilin was investigated in the mouse brain after kainic acid-induced seizure and seizure-induced hippocampal neuronal cell culture system using immunostaining analysis. Calsenilin was markedly decreased not only in the damaged cortex and CA3 region of hippocampus at 24 h after kainic acid-induced seizure but also in a cell-culture model of seizure like activity. In addition, immunoreactivity of calsenilin in the hippocampus derived from human epilepsy patient was significantly decreased compared with normal brain. These results demonstrate that the reduced expression of calsenilin may functionally be associated with the pathophysiology of status epilepticus. PMID- 12648753 TI - Early gamma response in human neuroelectric activity is correlated with neuropsychological test scores. AB - The goal of this study was to investigate whether early, time-locked 'sensory' gamma band response is correlated with a set of scores derived from neuropsychological tests. Neuroelectric responses were obtained under active auditory oddball paradigm. Of 67 reportedly healthy adults, 35 displayed time locked early gamma, G (+), and 24 did not, G (-). Out of 52 neuropsychological test scores, G (+) and G (-) groups differed on the basis of Wisconsin Card Sorting Test, Wechsler Memory Scale-Revised and Serial Digit Learning Test scores. Results of logistic regression analysis was statistically reliable (overall success rate of prediction: 93.33%). The results showed that early gamma response can be classified on the basis of neuropsychological test performance and is thus associated with higher cognitive functions, supporting the view that brain integrates bottom-up with top-down processing. PMID- 12648754 TI - Estradiol alters afterdischarge threshold and acquisition of amygdala kindled seizures in male rats. AB - We have previously shown that estradiol (E(2)) can initially increase and then decrease kindle seizure parameters in amygdala kindled male rats. This study focuses on the effects of estradiol benzoate (EB) on afterdischarge (AD) threshold and electrical kindling acquisition in intact male rats. After implantation of tripolar and monopolar electrodes in amygdala and dura surface respectively, effects of EB on AD threshold and electrical kindling acquisition were investigated by daily injection of EB (microg/kg) or sesame oil (EB solvent) in different groups of male rats. AD threshold was reduced significantly 0.25 h post EB treatment and reached to the lowest value after 24 h and remained almost constant at low values. Also, the number of trials for stage 5 (S(5)) and cumulative seconds of AD to complete kindling decreased significantly, when compared to rats without EB treatment. These results support a marked influence of E(2) on seizure process and convulsive pattern in the brain of male rats. Base on the previous reports about female rats and our findings, these E(2) effects are probably not sex dependent. PMID- 12648755 TI - Increased expression of 3 beta-hydroxysteroid dehydrogenase mRNA in dorsal root ganglion neurons of adult rats following peripheral nerve injury. AB - 3beta-hydroxysteroid dehydrogenase (3beta-HSD) is an enzyme that converts pregnenolone to progesterone. It has been believed that 3beta-HSD is simply a converting enzyme of female steroid hormone. Recently, 3beta-HSD expressing cells were identified in the spinal cord. Steroid synthesis in the nervous system may indicate that steroid plays a role in the nervous system. We report here the increased expression of 3beta-HSD mRNA in the dorsal root ganglion (DRG) after peripheral nerve injury using reverse transcription-polymerase chain reaction and in situ hybridization histochemistry techniques. We detected only a few 3beta-HSD signals in the naive DRG, and found that 3beta-HSD mRNA expression increased 3 days after injury and this increase was still observed at 14 days. Our results suggest that progesterone may have a role in the process against neuronal injury or in regeneration in the peripheral nervous system. PMID- 12648756 TI - Intravenous glucose infusion decreases intracisternal thyrotropin-releasing hormone induced vagal stimulation of gastric acid secretion in anesthetized rats. AB - Gastroparesis is a common complication of diabetes attributed to autonomic neuropathy. This study investigated whether acute hyperglycemia influences central thyrotropin-releasing hormone (TRH), a well-established brain medullary vagal stimulus, induced gastric acid secretion in overnight fasted, urethane anesthetized rats. Intravenous infusion of D-glucose (20%, 30% and 40%) dose dependently reduced intracisternal TRH-induced gastric acid secretion (71+/-28 micromol/90 min) by 39%, 90% and 100% respectively. Pretreatment with cholecystokinin(A) (CCK(A)) receptor antagonist devazepide (1 mg/kg) did not influence the inhibitory effect of intravenous glucose (30%). These results indicate that hyperglycemia may have a central effect to antagonize medullary TRH stimulation of vagal outflow to the stomach. PMID- 12648758 TI - Disruption of early event-related theta synchronization of human EEG in alexithymics viewing affective pictures. AB - The 62-channel EEG was recorded while control non-alexithymic (n=21) and alexithymic (n=20) participants viewed sequentially presented neutral, pleasant and unpleasant pictures and subjectively rated them after each presentation. The event-related synchronization (ERS) to these stimuli was assessed in the theta-1 (4-6 Hz) and theta-2 (6-8 Hz) frequency bands. The obtained findings indicate that alexithymia influences perception of only emotional stimuli. Over anterior cortical regions alexithymia vs. control individuals in response to both pleasant and unpleasant stimuli manifested decreased left hemisphere ERS in the early test period of 0-200 ms along with enhanced ERS in response to negative vs. positive and neutral stimuli in the right hemisphere at 200-600 ms after stimulus onset. The findings provide the first EEG evidence that alexithymia construct, associated with a cognitive deficit in initial evaluation of emotion, is indexed by disrupted early frontal synchronization in the upper theta band that can be best interpreted to reflect disregulation during appraisal of emotional stimuli. PMID- 12648757 TI - Disulfonic stilbenes prevent beta-amyloid (25-35) neuronal toxicity in rat cortical cultures. AB - Anion exchange proteins were recently identified among some of the proteins found clustered together in the hallmark plaques and tangles of Alzheimer's patient's brains. Anion exchange proteins underlie chloride/bicarbonate exchange, cell shape regulation and participate in removal of aged cells by the immune system. In this study we compared the neuroprotective efficacy of an anion exchanger inhibitor, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), against beta-amyloid((25-35)) neurotoxicity, staurosporine-induced apoptosis and glutamate-induced necrosis in primary cortical cultures. We demonstrate potent neuroprotective efficacy with DIDS against beta-amyloid((25-35)) and staurosporine, but not against glutamate. Our results suggest that anion exchange proteins may play an important role in beta-amyloid toxicity and that DIDS may represent a viable therapeutic agent for Alzheimer's disease. PMID- 12648759 TI - Comparison of response characteristics of cutaneous mechanoreceptors in normal and neuropathic Sprague-Dawley and spontaneously hypertensive rats. AB - The activity of single myelinated afferents was recorded from dorsal roots L4-5 in normal Spontaneously Hypertensive rats (SHR) and animals that developed mechanical hypersensitivity following ischemic injury to the sciatic nerve. Control and neuropathic SHRs had significantly higher paw withdrawal threshold to mechanical stimulation than control and neuropathic Sprague-Dawleys (SD). In the SHR rats the mechanical response properties of afferents conducting through the injury site were similar to normals and many of the afferents not conducting through the injury site had spontaneous activity. The only significant difference between the two strains was a faster conduction velocity in afferents recorded from SHR than SD rats. Thus, the behavioral hyposensitivity and less development of mechanical allodynia of SHR rats, compared to SD is not due to differences in the properties of myelinated afferents, but probably involves differences in central inhibitory mechanisms in the two strains. PMID- 12648760 TI - Blockade of bombesin-like peptide receptors impairs inhibitory avoidance learning in mice. AB - Several studies reported that peripheral administration of bombesin (BN) and gastrin-releasing peptide (GRP) improved some forms of memory performance. In the present study, we examined the role of endogenous BN-like peptide(s) for the acquisition of inhibitory avoidance learning in mice using BN-like peptide receptor antagonists. An administration of [Leu(13)-(psi-CH(2)NH)-Leu(14)]BN (antagonizes GRP-R>neuromedin B receptor (NMB-R)) impaired the performance of inhibitory avoidance learning in all doses (16, 32, 64 nmol/kg). While the effect was somewhat lesser than [Leu(13)-(psi-CH(2)NH)-Leu(14)]BN, BIM23127 (antagonizes NMB-R>GRP-R) also impaired performance in a moderate dose (32 nmol/kg). These results showed that endogenous BN-like peptides have some role(s) for the modulation of learning and memory, and suggest that NMB/NMB-R pathway may also be involved in the memory acquisition and modulation as well as GRP/GRP-R pathway. PMID- 12648761 TI - Increased frequency of a new polymorphism in the cell division cycle 2 (cdc2) gene in patients with Alzheimer's disease and frontotemporal dementia. AB - Recent studies show linkage between Alzheimer's disease (AD) and two loci on chromosome 10. The cell division cycle 2 (cdc2) gene is located close to one of the chromosome 10 markers, and is a candidate gene for AD since it is involved in the pathogenesis of AD. We sequenced coding exons and flanking intronic sequences and the promoter region on the cdc2 gene and found three new single nucleotide polymorphisms (SNPs). We analyzed 272 Caucasian AD cases, 160 controls and 70 cases with frontotemporal dementia (FTD) for these SNPs. Homozygosity for one of the SNPs (Ex6+7I/D) was more frequent in both AD and FTD cases than in controls. In the combined tauopathy (AD and FTD) group the odds ratio (OR) was 1.77 (95% CI 1.19-2.63) for the Ex6+7II genotype. Our findings suggest that the Ex6+7I allele is associated with tauopathies, both AD and FTD. PMID- 12648762 TI - Identifying the components of a postsynaptic potential and their amplitude, latency and shape fluctuations: analysis by means of autocovariance functions and a stochastic infinite cable model. AB - In addition to amplitude fluctuations, physiological mechanisms may introduce latency and shape fluctuations in the components of a postsynaptic potential (PSP). Latency fluctuations may be originated mainly by presynaptic factors. Shape fluctuations may be produced by changes in the background synaptic activity received by the postsynaptic neuron, which affect the cell membrane resistance. This article aims to develop a unified approach for the analysis of amplitude, latency and shape fluctuations in the components of a PSP. The analysis is based on: (i) the Autocovariance Functions of the PSP (ACOVs); (ii) a mathematical model able to predict the average and ACOVs of a PSP with specified components and fluctuations (the 'Stochastic Infinite Cable Model' (SICM)); and (iii) a procedure to estimate the SICM parameters that best reproduce the average and ACOVs of a given PSP (the 'SICM-based PSP identification procedure' (SICM-IP)). The SICM-IP is tested with simulated PSPs. The results obtained support the feasibility of the approach. PMID- 12648763 TI - The use of time-variant EEG Granger causality for inspecting directed interdependencies of neural assemblies. AB - Understanding of brain functioning requires the investigation of activated cortical networks, in particular the detection of interactions between different cortical sites. Commonly, coherence and correlation are used to describe interrelations between EEG signals. However, on this basis, no statements on causality or the direction of their interrelations are possible. Causality between two signals may be expressed in terms of upgrading the predictability of one signal by the knowledge of the immediate past of the other signal. The best established approach in this context is the so-called Granger causality. The classical estimation of Granger causality requires the stationarity of the signals. In this way, transient pathways of information transfer stay hidden. The study presents an adaptive estimation of Granger causality. Simulations demonstrate the usefulness of the time-variant Granger causality for detecting dynamic causal relations within time intervals of less than 100 ms. The time variant Granger causality is applied to EEG data from the Stroop task. It was shown that conflict situations generate dense webs of interactions directed from posterior to anterior cortical sites. The web of directed interactions occurs mainly 400 ms after the stimulus onset and lasts up to the end of the task. PMID- 12648764 TI - Differential tissue shrinkage and compression in the z-axis: implications for optical disector counting in vibratome-, plastic- and cryosections. AB - The optical disector is among the most efficient cell counting methods, but its accuracy depends on an undistorted particle distribution in the z-axis of tissue sections. Because the optical disector samples particle densities exclusively in the center of sections, it is essential for unbiased estimates of particle numbers that differential shrinkage or compression (and resulting differences in particle densities along the z-axis) are known and corrected. Here we examined, quantified, and compared differential shrinkage and compression of vibratome-, celloidin- and cryosections. Vibratome sections showed a significant z-axis distortion, while celloidin- and cryosections were minimally distorted. Results were directly compared with previous data obtained from paraffin and methacrylate sections. We conclude that z-axis distortion varies significantly between embedding and sectioning methods, and that vibratome-, methacrylate- and paraffin sections can result in grossly biased estimates. We describe a simple method for assessing differential z-axis shrinkage or compression, as well as simple strategies to minimize the bias of the optical disector. Minimal bias can be achieved by either adjusting the placement and extent of counting boxes and guard spaces for sampling, or by applying a correction factor in cases when guard spaces are deemed essential for particle recognition. PMID- 12648765 TI - Production of dopaminergic neurons for cell therapy in the treatment of Parkinson's disease. AB - Dopaminergic cell therapy is a potential viable treatment for Parkinson's disease. However, lack of a well-characterized cell preparation of known phenotypic composition containing a high percentage of dopaminergic neurons, has prevented a definitive, controlled, pilot clinical trial from being conducted. We report the successful in vitro expansion of rat E12 mesencephalic progenitors to produce 5-fold the normal number of dopaminergic neurons. The expanded neurons (MAP2+) were detached, resuspended, and formed into small aggregates of 10-200 neurons containing 25-50% of dopaminergic neurons (TH+) that will likely be optimal for use in successful cell therapy. After storage in DPBS, in 0 mM Ca(2+) for up to 24 h at room temperature, aggregated cells were still 90% viable. These results demonstrate that it might be feasible to use a similar protocol to expand human dopaminergic progenitors in vitro. If successful, the requisite large numbers of dopaminergic neurons required to conduct a pilot clinical trial for Parkinson's disease will be produced in vitro. Indications are that the cells can be maintained at optimal viability for the duration of the neural transplantation procedure, under real operating conditions. PMID- 12648766 TI - Codon optimization of Caenorhabditis elegans GluCl ion channel genes for mammalian cells dramatically improves expression levels. AB - Organisms use synonymous codons in a highly non-random fashion. These codon usage biases sometimes frustrate attempts to express high levels of exogenous genes in hosts of widely divergent species. The Caenorhabditis elegans GluClalpha1 and GluClbeta genes form a functional glutamate and ivermectin-gated chloride channel when expressed in Xenopus oocytes, but expression is weak in mammalian cells. We have constructed synthetic genes that retain the amino acid sequence of the wild type GluCl channel proteins, but use codons that are optimal for mammalian cell expression. We have tagged the native and codon-optimized GluCl cDNAs with enhanced yellow fluorescent protein (EYFP, GluClalpha1 subunit) and enhanced cyan fluorescent protein (EFCP, GluClbeta subunit), expressed the channels in E18 rat hippocampal neurons and measured the relative expression levels of the two genes with fluorescence microscopy as well as with electrophysiology. Codon optimization provides a 6- to 9-fold increase in expression, allowing the conclusions that the ivermectin-gated channel has an EC(50) of 1.2 nM and a Hill coefficient of 1.9. We also confirm that the Y182F mutation in the codon optimized beta subunit results in a heteromeric channel that retains the response to ivermectin while reducing the response to 100 microM glutamate by 7-fold. The engineered GluCl channel is the first codon-optimized membrane protein expressed in mammalian cells and may be useful for selectively silencing specific neuronal populations in vivo. PMID- 12648767 TI - Novel functional imaging technique from brain surface with optical coherence tomography enabling visualization of depth resolved functional structure in vivo. AB - Mapping of the activity of brain by optical intrinsic signal imaging (OISI) provides a two-dimensional activation pattern of visual cortical areas at a resolution of a few hundred microns. However, integration of the intrinsic signal over depth results in loss of finer information about functional organization across the depth. Here, we report the first successful implementation of optical coherence tomography (OCT) at around 30 microm depth resolution to investigate cortical functions of a cat brain in vivo. This technique, named functional OCT (fOCT) provided visually evoked changes in the OCT signal. The fOCT signal shows stimulus specificity that correlates well with that of the intrinsic signals and provides depth resolved layer specific functional information. PMID- 12648768 TI - A robust statistics-based global energy function for the alignment of serially acquired autoradiographic sections. AB - Autoradiographic analysis of the functional changes occurring in the rat brain are most often performed on coronal sections that allow a good insight into the events occurring at the structural level but lacks the 3D context which is necessary to fully understand the involvement of the brain structures in specific situations like focal seizures with or without generalization. Therefore a robust, fully-automated algorithm for the registration of serially acquired autoradiographic sections is presented. The method accounts for the main difficulties of autoradiographic alignment: corrupted data (cuts and tears), dissimilarities or discontinuities between slices, non parallel or missing slices. The approach relies on the minimization of a global energy function based on robust statistics. The energy function measures the similarity between a slice and its neighborhood in the 3D volume. No particular direction is privileged in the method, so that global offsets, biases in the estimation or error propagations are avoided. The method is evaluated qualitatively and quantitatively on real autoradiographic data. Rat brain autoradiographic volumes are reconstructed with registration errors less than 1 degree in rotation and less than 1 pixel in translation. PMID- 12648769 TI - Misallocation of variance in event-related potentials: simulation studies on the effects of test power, topography, and baseline-to-peak versus principal component quantifications. AB - Since Wood and McCarthy's simulation study (Electroenceph Clin Neurophysiol 1984;59:249-260), the use of principal component analysis (PCA) as a tool for the identification and quantification of event-related potentials (ERP) has been considered a challenge. Three relevant aspects have not been fully acknowledged in previous studies, however, and were therefore investigated in the present simulation study. Firstly, the impact of test power on the amount of variance misallocation was studied. Secondly, the impact of ERP component topography on variance misallocation was investigated. Thirdly, a systematic evaluation of variance misallocation in baseline-to-peak derived ERP measures was performed. Results based on an overall set of 2700 simulations indicate that: (a) variance misallocation is reduced to an almost acceptable level when an appropriate test power is simulated; (b) the overall amount of variance misallocation remains at an almost acceptable level when systematic topographic effects are simulated in combination with an appropriate test power; and (c) variance misallocation is in fact also a problem in baseline-to-peak measures. These findings confirm that, when used appropriately, PCA is a helpful and efficient tool for the identification and quantification of ERPs. PMID- 12648771 TI - Condensed complexes of cholesterol and phospholipids. AB - There is overwhelming evidence that lipid bilayer regions of animal cell membranes are in a liquid state. Quantitative models of these bilayer regions must then be models of liquids. These liquids are highly non-ideal. For example, it has been known for more than 75 years that mixtures of cholesterol and certain phospholipids undergo an area contraction or condensation in lipid monolayers at the air-water interface. In the past 3 years, a thermodynamic model of "condensed complexes" has been proposed to account for this non-ideal behavior. Here we give an overview of the model, its relation to other models, and to modern views of the properties of animal cell membranes. PMID- 12648772 TI - Role of cholesterol in lipid raft formation: lessons from lipid model systems. AB - Biochemical and cell-biological experiments have identified cholesterol as an important component of lipid 'rafts' and related structures (e.g., caveolae) in mammalian cell membranes, and membrane cholesterol levels as a key factor in determining raft stability and organization. Studies using cholesterol-containing bilayers as model systems have provided important insights into the roles that cholesterol plays in determining lipid raft behavior. This review will discuss recent progress in understanding two aspects of lipid-cholesterol interactions that are particularly relevant to understanding the formation and properties of lipid rafts. First, we will consider evidence that cholesterol interacts differentially with different membrane lipids, associating particularly strongly with saturated, high-melting phospho- and sphingolipids and particularly weakly with highly unsaturated lipid species. Second, we will review recent progress in reconstituting and directly observing segregated raft-like (liquid-ordered) domains in model membranes that mimic the lipid compositions of natural membranes incorporating raft domains. PMID- 12648773 TI - Phospholipid/cholesterol model membranes: formation of cholesterol crystallites. AB - Experimental data that define conditions under which cholesterol crystallites form in cholesterol/phospholipid model membranes are reviewed. Structural features of the phospholipids that determine cholesterol crystallization include the length and degree of unsaturation of the acyl chains, the presence of charge on the headgroups and interheadgroup hydrogen bonds. PMID- 12648774 TI - Direct evidence for cholesterol crystalline domains in biological membranes: role in human pathobiology. AB - This review will discuss the use of small-angle X-ray diffraction approaches to study the organization of lipids in plasma membranes derived from two distinct mammalian cell types: arterial smooth muscle cells and ocular lens fiber cells. These studies indicate that cholesterol at an elevated concentration can self associate and form immiscible domains in the plasma membrane, a phenomenon that contributes to both physiologic and pathologic cellular processes, depending on tissue source. In plasma membrane samples isolated from atherosclerotic smooth muscle cells, the formation of sterol-rich domains is associated with loss of normal cell function, including ion transport activity and control of cell replication. Analysis of meridional diffraction patterns from intact and reconstituted plasma membrane samples indicates the presence of an immiscible cholesterol domain with a unit cell periodicity of 34 A, consistent with a cholesterol monohydrate tail-to-tail bilayer, under disease conditions. These cholesterol domains were observed in smooth muscle cells enriched with cholesterol in vitro as well as from cells obtained ex vivo from an animal model of atherosclerosis. By contrast, well-defined cholesterol domains appear to be essential to the normal physiology of fiber cell plasma membranes of the human ocular lens. The organization of cholesterol into separate domains underlies the role of lens fiber cell plasma membranes in maintaining lens transparency. These domains may also interfere with cataractogenic aggregation of soluble lens proteins at the membrane surface. Taken together, these analyses provide examples of both physiologic and pathologic roles that sterol-rich domains may have in mammalian plasma membranes. These findings support a model of the membrane in which cholesterol aggregates into structurally distinct regions that regulate the function of the cell membrane. PMID- 12648775 TI - Structural information about organized cholesterol domains from specific antibody recognition. AB - Cholesterol-rich domains have been observed to exist in cell membranes under physiological and pathological conditions. Their compositions and the microenvironment of their formation vary over a wide range. Very little information is however available on the molecular structure and organization of these domains. The techniques available to provide such structural information are reviewed here first. The possibility of using tailor-made antibodies as reporters of molecular organization in membranes is then considered. The concept of antibodies recognizing molecular organization rather than single molecular epitopes is established, reviewing the existing works on antibody and protein recognition of crystalline molecular arrays. The information that such antibodies could provide in cells is finally examined together with a proof of application. PMID- 12648776 TI - Relationship between cholesterol trafficking and signaling in rafts and caveolae. AB - Caveolae and lipid rafts are two distinct populations of free cholesterol, sphingolipid (FC/SPH)-rich cell surface microdomains. They differ in stability, shape, and the presence or absence of caveolin (present in caveolae) or GPI anchored proteins (enriched in lipid rafts). In primary cells, caveolae and rafts support the assembly of different signaling complexes, though signal transduction from both is strongly dependent on the presence of FC. It was initially thought that FC promoted the formation of inactive reservoirs of signaling proteins. Recent data supports the concept of a more dynamic role for FC in caveolae and probably, also lipid rafts. It is more likely that the FC content of these domains is actively modulated as protein complexes are formed and, following signal transduction, disassembled. In transformed cell lines with few caveolae, little caveolin and a preponderance of rafts, complexes normally assembled on caveolae may function in rafts, albeit with altered kinetics. However, caveolae and lipid rafts appear not to be interconvertible. The presence of non-caveolar pools of caveolin in recycling endosomes (RE), the trans-Golgi network (TGN) and in mobile chaperone complexes is now recognized. A role in the uptake of microorganisms by cells ascribed to caveolae now seems more likely to be mediated by cell surface rafts. PMID- 12648777 TI - Dynamics of raft molecules in the cell and artificial membranes: approaches by pulse EPR spin labeling and single molecule optical microscopy. AB - Lipid rafts in the plasma membrane, domains rich in cholesterol and sphingolipids, have been implicated in a number of important membrane functions. Detergent insolubility has been used to define membrane "rafts" biochemically. However, such an approach does not directly contribute to the understanding of the size and the lifetime of rafts, dynamics of the raft-constituent molecules, and the function of rafts in the membrane in situ. To address these issues, we have developed pulse EPR spin labeling and single molecule tracking optical techniques for studies of rafts in both artificial and cell membranes. In this review, we summarize our results and perspectives obtained by using these methods. We emphasize the importance of clearly distinguishing small/unstable rafts (lifetime shorter than a millisecond) in unstimulated cells and stabilized rafts induced by liganded and oligomerized (GPI-anchored) receptor molecules (core receptor rafts, lifetime over a few minutes). We propose that these stabilized rafts further induce temporal, greater rafts (signaling rafts, lifetime on the order of a second) for signaling by coalescing other small/unstable rafts, including those in the inner leaflet of the membrane, each containing perhaps one molecule of the downstream effector molecules. At variance with the general view, we emphasize the importance of cholesterol segregation from the liquid-crystalline unsaturated bulk-phase membrane for formation of the rafts, rather than the affinity of cholesterol and saturated alkyl chains. In the binary mixture of cholesterol and an unsaturated phospholipid, cholesterol is segregated out from the bulk unsaturated liquid-crystalline phase, forming cholesterol-enriched domains or clustered cholesterol domains, probably due to the lateral nonconformability between the rigid planar transfused ring structure of cholesterol and the rigid bend of the unsaturated alkyl chain at C9-C10. However, such cholesterol-rich domains are small, perhaps consisting of only several cholesterol molecules, and are short-lived, on the order of 1-100 ns. We speculate that these cholesterol-enriched domains may be stabilized by the presence of saturated alkyl chains of sphingomyelin or glycosphingolipids, and also by clustered raft proteins. In the influenza viral membrane, one of the simplest forms of a biological membrane, the lifetime of a protein and cholesterol-rich domain was evaluated to be on the order of 100 micro, again showing the short lifetime of rafts in an unstimulated state. Finally, we propose a thermal Lego model for rafts as the basic building blocks for signaling pathways in the plasma membrane. PMID- 12648778 TI - Lipid rafts are critical membrane domains in blood platelet activation processes. AB - Among the various hematopoi;etic cells, platelets are critical for maintaining the integrity of the vascular system. They must be rapidly activated by sequential and coordinated mechanisms in order to efficiently prevent haemorrhage upon vascular injury. Several signal transduction pathways lead to platelet activation in vitro and in vivo, among them, several are initiated via receptors or co-receptors containing immuno-receptor tyrosine-based activation motifs (ITAM) which trigger downstream signalling like the immune receptors in lymphocytes. However, in contrast to immune cells for which the role of lipid rafts in signalling has largely been described, the involvement of laterally segregated membrane microdomains in platelet activation has been investigated only recently. The results obtained until now strongly suggest that early steps of platelet activation via the collagen receptor GpVI or via FcgammaRIIa occur preferentially in these microdomains where specific proteins efficiently organize key downstream signalling pathways. In addition, lipid rafts also contribute to platelet activation via heterotrimeric G-protein-coupled receptors. They are sites where the phosphoinositide (PI) metabolism is highly active, leading to a local generation of lipid second messengers such as phosphatidylinositol 3,4,5 trisphosphate. Here, evidence is accumulating that cholesterol-enriched membrane microdomains are part of a general process that contributes to the efficiency and the coordination of platelet activation mechanisms. Here we will discuss the biochemical and functional characterizations of human platelet rafts and their potential impact in platelet physiopathology. PMID- 12648779 TI - Molecular characterization of the detergent-insoluble cholesterol-rich membrane microdomain (raft) of the central nervous system. AB - Many fundamental neurological issues such as neuronal polarity, the formation and remodeling of synapses, synaptic transmission, and the pathogenesis of the neuronal cell death are closely related to the membrane dynamics. The elucidation of functional roles of a detergent-insoluble cholesterol-rich domain (raft) could therefore provide good clues to the molecular understanding of these important phenomena, for the participation of the raft in the fundamental cell functions, such as signal transduction and selective transport of lipids and proteins, has been elucidated in nonneural cells. Interestingly, the brain is rich in raft and the brain-derived raft differs in its lipid and protein components from other tissue-derived rafts. Since many excellent reviews are written on the membrane lipid dynamics of this microdomain, signal transduction, and neuronal glycolipids, we review on the characterization of the raft proteins recovered in the detergent-insoluble low-density fraction from rat brain. Special focus is addressed on the biochemical characterization of a neuronal enriched protein, NAP 22, for the lipid organizing activity of this protein has become increasingly clear. PMID- 12648780 TI - Role of cholesterol in synapse formation and function. AB - Cholesterol is a multifaceted molecule, which serves as essential membrane component, as cofactor for signaling molecules and as precursor for steroid hormones. Consequently, defects in cholesterol metabolism cause devastating diseases. So far, the role of cholesterol in the nervous system is less well understood. Recent studies showed that cultured neurons from the mammalian central nervous system (CNS) require glia-derived cholesterol to form numerous and efficient synapses. This suggests that the availability of cholesterol in neurons limits the extent of synaptogenesis. Here, I will summarize the experimental evidence for this hypothesis, describe what is known about the structural and functional role of cholesterol at synapses, and discuss how cholesterol may influence synapse development and stability. PMID- 12648781 TI - Amyloid beta-protein interactions with membranes and cholesterol: causes or casualties of Alzheimer's disease. AB - Amyloid beta-protein (Abeta) is thought to be one of the primary factors causing neurodegeneration in Alzheimer's disease (AD). This protein is an amphipathic molecule that perturbs membranes, binds lipids and alters cell function. Several studies have reported that Abeta alters membrane fluidity but the direction of this effect has not been consistently observed and explanations for this lack of consistency are proposed. Cholesterol is a key component of membranes and cholesterol interacts with Abeta in a reciprocal manner. Abeta impacts on cholesterol homeostasis and modification of cholesterol levels alters Abeta expression. In addition, certain cholesterol lowering drugs (statins) appear to reduce the risk of AD in human subjects. However, the role of changes in the total amount of brain cholesterol in AD and the mechanisms of action of statins in lowering the risk of AD are unclear. Here we discuss data on membranes, cholesterol, Abeta and AD, and propose that modification of the transbilayer distribution of cholesterol in contrast to a change in the total amount of cholesterol provides a cooperative environment for Abeta synthesis and accumulation in membranes leading to cell dysfunction including disruption in cholesterol homeostasis. PMID- 12648782 TI - Tumour therapy with radionuclides: assessment of progress and problems. AB - Radionuclide therapy is a promising modality for treatment of tumours of haematopoietic origin while the success for treatment of solid tumours so far has been limited. The authors consider radionuclide therapy mainly as a method to eradicate disseminated tumour cells and small metastases while bulky tumours and large metastases have to be treated surgically or by external radiation therapy. The promising therapeutic results for haematological tumours give hope that radionuclide therapy will have a breakthrough also for treatment of disseminated cells from solid tumours. New knowledge related to this is continuously emerging since new molecular target structures are being characterised and the knowledge on pharmacokinetics and cellular processing of different types of targeting agents increases. There is also improved understanding of the factors of importance for the choice of appropriate radionuclides with respect to their decay properties and the therapeutic applications. Furthermore, new methods to modify the uptake of radionuclides in tumour cells and normal tissues are emerging. However, we still need improvements regarding dosimetry and treatment planning as well as an increased knowledge about the tolerance doses for normal tissues and the radiobiological effects on tumour cells. This is especially important in targeted radionuclide therapy where the dose rates often are lower than 1Gy/h. PMID- 12648783 TI - First results of a phase I/II dose escalation trial in non-small cell lung cancer using three-dimensional conformal radiotherapy. AB - PURPOSE: To evaluate the feasibility of dose escalation in non-small cell lung cancer (NSCLC) using three-dimensional conformal radiation therapy. PATIENTS AND METHODS: The main eligibility criteria of the trial were: pathologically proven inoperable NSCLC, ECOG performance status or=grade 3 (SWOG), grade 3 early and grade 2 late esophageal toxicity or any other (RTOG) grade 3 or 4 complications). RESULTS: Fifty-five patients were included. Tumor stage was I/II in 47%, IIIA in 33% and IIIB in 20%. The majority of the patients received a dose of 74.3 Gy (n=17) or 81.0 Gy (n=23). Radiation pneumonitis occurred in seven patients: four patients developed a grade 2, two patients grade 3 and one patient a grade 4. Esophageal toxicity was mild. In 50 patients tumor response at 3 months follow-up was evaluable. In six patients a complete response was recorded, in 38 a partial response, five patients had stable disease and one patient experienced progressive disease. Only one patient developed an isolated failure in an uninvolved nodal area. So far the radiation dose was safely escalated to 87.8 Gy in group 1 (lowest rMLD), 81.0 Gy in groups 2 and 3 and 74.3 Gy in group 4. CONCLUSION: Three-dimensional conformal radiotherapy enables significant dose escalation in NSCLC. The maximum tolerable dose has not yet been reached in any risk group. PMID- 12648785 TI - Impact of target reproducibility on tumor dose in stereotactic radiotherapy of targets in the lung and liver. AB - BACKGROUND AND PURPOSE: Previous analyses of target reproducibility in extracranial stereotactic radiotherapy have revealed standard security margins for planning target volume (PTV) definition of 5mm in axial and 5-10mm in longitudinal direction. In this study the reproducibility of the clinical target volume (CTV) of lung and liver tumors within the PTV over the complete course of hypofractionated treatment is evaluated. The impact of target mobility on dose to the CTV is assessed by dose-volume histograms (DVH). MATERIALS AND METHODS: Twenty-two pulmonary and 21 hepatic targets were treated with three stereotactic fractions of 10 Gy to the PTV-enclosing 100%-isodose with normalization to 150% at the isocenter. A conformal dose distribution was related to the PTV, which was defined by margins of 5-10mm added to the CTV. Prior to each fraction a computed tomography (CT)-simulation over the complete target volume was performed resulting in a total of 60 CT-simulations for lung and 58 CT-simulations for hepatic targets. The CTV from each CT-simulation was segmented and matched with the CT-study used for treatment planning. A DVH of the simulated CTV was calculated for each fraction. The target coverage (TC) of dose to the simulated CTV was defined as the proportion of the CTV receiving at least the reference dose (100%). RESULTS: A decrease of TC to <95% was found in 3/60 simulations (5%) of pulmonary and 7/58 simulations (12%) of hepatic targets. In two of 22 pulmonary targets (9%) and in four of 21 hepatic targets (19%) a TC of <95% occurred in at least one fraction. At risk for a decreased TC <95% were pulmonary targets with increased breathing mobility and hepatic targets with a CTV exceeding 100 cm(3). CONCLUSIONS: Target reproducibility was precise within the reference isodose in 91% of lung and 81% of liver tumors with a TC of the complete CTV >or=95% at each fraction of treatment. Pulmonary targets with increased breathing mobility and liver tumors >100 cm(3) are at risk for target deviation exceeding the standard security margins for PTV-definition at least for one fraction and require individual evaluation of sufficient margins. PMID- 12648784 TI - Carbon ion radiotherapy for stage I non-small cell lung cancer. AB - BACKGROUND AND PURPOSE: Heavy ion radiotherapy is a promising modality because of its excellent dose localization and high biological effect on tumors. Using carbon beams, a dose escalation study was conducted for the treatment of stage I non-small cell lung cancer (NSCLC) to determine the optimal dose. MATERIALS AND METHODS: The first stage phase I/II trial using 18 fractions over 6 weeks for 47 patients and the second one using nine fractions over 3 weeks for 34 patients were conducted by the dose escalation method from 59.4 to 95.4 Gray equivalents (GyE) in incremental steps of 10% and from 68.4 to 79.2 GyE in 5% increments, respectively. The local control and survival rates were obtained using the Kaplan Meier method. RESULTS: Radiation pneumonitis at grade III occurred in three of 81 patients, but they fully recovered. This was not a dose-limiting factor. The local control rates in the first and second trials were 64% and 84%, respectively. The total recurrence rate in both trials was 23.2%. The infield local recurrence in the first trial was significantly dependent on carbon dose. The doses greater than 86.4 GyE at 18 fractions and 72 GyE at nine fractions achieved a local control of 90% and 95%, respectively. The 5 year overall and cause-specific survivals in 81 patients were 42% and 60%, respectively. CONCLUSIONS: With our dose escalation study, the optimum safety and efficacy dose of carbon beams was determined. Carbon beam therapy attained almost the same results as surgery for stage I NSCLC although this was a I/II study. PMID- 12648786 TI - Neoadjuvant androgen deprivation and prostate gland shrinkage during conformal radiotherapy. AB - PURPOSE: The shrinking effect of 3-month neoadjuvant androgen deprivation (NAD) on preradiotherapy prostate gland volume is well documented. However, recently, it has been shown that the cancerous prostate gland keeps shrinking up to 12 months after NAD start. Thus, if such a reduction is not taken into account, a larger than planned portion of the surrounding normal tissues might shift in the high-dose region during conformal radiotherapy (3DCRT) course. The present study was undertaken to quantify this issue. MATERIALS AND METHODS: Prostate gland volume reduction between planning CT (plCT) and the last week of 3DCRT (tmtCT) was prospectively assessed in 33 consecutive patients with localized prostate carcinoma. The median time interval between plCT and tmtCT was 2.5 months (2.1 2.7 months). A single observer was asked to draw on each slice prostate gland volume as appropriate. The observer was 'blind' to the timing of CT (plCT vs. tmtCT). In order to estimate intra-observer variability, prostate gland delineation was repeated twice for each data set. Mean prostate gland change, plCT and tmtCT cumulative dose volume histogram (DVH) calculations for the rectum were analyzed for each patient. Results were correlated to AD status and its duration before plCT. Means were compared by non-parametric rank tests. RESULTS: Based on an internal protocol, 14 patients (42%) did not receive AD, while 19 patients (58%) had undergone neoadjuvant and concomitant AD. The median duration of AD before plCT ranged from 0.2 to 6 months (median: 2.9 months). Although individual data were highly variable, compared to plCT volume, mean prostate gland volume change at the end of 3DCRT was similar for patients receiving ( 7.3%) or not (-7%) androgen deprivation (P=0.77). However, within the group of patients treated with hormones, patients starting AD within 3 months from plCT had a significantly larger reduction in prostate volume (-14.2%) than patients with longer NAD duration (-1.1%, P=0.03). At tmtCT, on average, patients undergoing 3DCRT within 3 months from AD start showed an increase of the amount of rectum receiving 40-75 Gy compared to plCT values. At 40 Gy (V40) the mean difference between tmtCT and plCT was +7.5%. In the other two groups, average variations of V40-70 were within +/-2% of plCT values. However, these differences are not significant. CONCLUSION: For patients who undergo plCT and 3DCRT shortly after AD start, prostate gland shrinkage may be substantial. In some of these patients, this might lead to an unexpected increase of the percentage of rectal wall exposed to intermediate doses. PMID- 12648787 TI - Predictive factors of acute urinary morbidity after iodine-125 brachytherapy for localised prostate cancer: a phase 2 study. AB - PURPOSE: To analyse predictive factors of acute urinary morbidity after transperineal permanent prostate brachytherapy. METHODS AND MATERIALS: Sixty patients treated in a phase 2 study with iodine-125 brachytherapy (9/1998 to 2/2000) for localised prostate adenocarcinoma were analysed after at least 1-year follow-up. Prescribed dose was 144 Gy and all patients had a pre-planning and a post-implant dosimetry. Urinary morbidity was evaluated prospectively using the Radiation Therapy Oncology Group (RTOG) scale. We examined the relationship between pre-implant ultrasound prostate volume, post-implant CT-scan prostate volume, neoadjuvant hormonotherapy, total number of needles and seeds, post implant dosimetry variables, first 30 vs. last 30 treated patients and post implant urinary morbidity. RESULTS: All patients experienced some degree of urinary distress symptoms after treatment. Symptoms were generally mild grade 1 in 56% and grade 2 in 10% lasting less than 6 months. Eight patients (13%) required bladder catheter for acute urinary obstruction. At 1-year follow-up, nine patients (15%) complained from persistent dysuria requiring in three cases endoscopic prostate resection. The percentage of urethra volume receiving 216 Gy (cut-off 40%) and the pre-implant prostate volume (cut-off 31 ml) were the only statistically significant predictor of grade 2-3 or persistent urinary morbidity on multivariate analysis. CONCLUSION: Our short-term data suggest that both pre implant prostate volume value and post-implant V.U. 150 value might be predictors for urinary morbidity after prostate brachytherapy. PMID- 12648789 TI - Impact of target volume coverage with Radiation Therapy Oncology Group (RTOG) 98 05 guidelines for transrectal ultrasound guided permanent Iodine-125 prostate implants. AB - PURPOSE: Despite the wide use of permanent prostate implants for the treatment of early stage prostate cancer, there is no consensus for optimal pre-implant planning guidelines that results in maximal post-implant target coverage. The purpose of this study was to compare post-implant target volume coverage and dosimetry between patients treated before and after Radiation Therapy Oncology Group (RTOG) 98-05 guidelines were adopted using several dosimetric endpoints. MATERIALS AND METHODS: Ten consecutively treated patients before the adoption of the RTOG 98-05 planning guidelines were compared with ten consecutively treated patients after implementation of the guidelines. Pre-implant planning for patients treated pre-RTOG was based on the clinical target volume (CTV) defined by the pre-implant TRUS definition of the prostate. The CTV was expanded in each dimension according to RTOG 98-05 and defined as the planning target volume. The evaluation target volume was defined as the post-implant computed tomography definition of the prostate based on RTOG 98-05 protocol recommendations. Implant quality indicators included V(100), V(90), V(100), and Coverage Index (CI). RESULTS: The pre-RTOG median V(100), V(90), D(90), and CI values were 82.8, 88.9%, 126.5 Gy, and 17.1, respectively. The median post-RTOG V(100), V(90), D(90), and CI values were 96.0, 97.8%, 169.2 Gy, and 4.0, respectively. These differences were all statistically significant. CONCLUSIONS: Implementation of the RTOG 98-05 implant planning guidelines has increased coverage of the prostate by the prescription isodose lines compared with our previous technique, as indicated by post-implant dosimetry indices such as V(100), V(90), D(90). The CI was also improved significantly with the protocol guidelines. Our data confirms the validity of the RTOG 98-05 implant guidelines for pre-implant planning as it relates to enlargement of the CTV to ensure adequate margin between the CTV and the prescription isodose lines. PMID- 12648788 TI - Results of high dose rate afterloading brachytherapy boost to conventional external beam radiation therapy for initial and locally advanced prostate cancer. AB - PURPOSE: To evaluate the impact on biochemical control (bNED), acute and late gastro-intestinal (GI) and urological (GU) morbidity of initial and locally advanced prostate cancer treated with fractionated transrectal ultrasound-guided (TRUS) high dose rate after loading brachytherapy (HDR-B) as a boost to conventional external beam radiation therapy (EBRT). PATIENTS AND METHODS: From March 1997 to February 2000 a total of 119 patients with any of the following characteristics were eligible for study entry: biopsy proven adenocarcinoma Gleason scored (GS), initial prostatic specific antigen (PSA) level dosage 1992 AJCC clinical stage T3a or less, and prostatic volume <60 cc. All patients had prior to HDR-B a course of EBRT 6 MV photons to a median dose of 45 Gy, in 1.8 Gy fractions, to the prostate and seminal vesicles only. HDR-B treatment planning and dosimetric calculations were generated with the Nucletron Planning System. Patients were grouped into two groups, according to their risk for biochemical failure: low-risk group without (LR) or with neoadjuvant total androgen deprivation (AD) prior to EBRT (LR+AD) and high-risk group without (HR) or with neoadjuvant AD (HR+AD), for bNED and dose-escalation protocol. LR encompassed patients who presented GS<6, T1 or T2a and or initial PSA<10 ng/ml, who were treated with 16 Gy (4 Gy fractions, b.i.d.) HDR-B. The remaining patients were grouped into HR or HR+AD and received 20 Gy (5 Gy fractions, b.i.d.) HDR-B. The planning was optimized using the standard geometric optimization. Biological effective doses (BED) for tumor control and late responding tissue were calculated using a alpha/beta ratio of 1.5 and 3 Gy, respectively. They were matched with bNED, acute and late gastrointestinal (GI) and urological (GU) morbidity, according to the RTOG/EORTC scoring criteria. RESULTS: Median age of patients was 68 years (range 47-83), with a median follow-up of 41 months (range 18-48). The crude and actuarial biochemical controls (bNED) in 48 months for all patients were 69.5 and 75.3%, respectively. When grouped into LR, LR+AD, HR and HR+AD the actuarial bNED were 78.2, 76, 76 and 72.3% (P=0.89), respectively. Acute GU and GI morbidity G1-2 were seen in 18.5% (20/108) and 10.2% (11/108) of patients with spontaneous regression. Late GI and GU morbidity G1-2 were seen in 12% (13/108) and 4.6 (5/108) of patients, with no need of intervention. No acute or late G3-4 GU or GI morbidity was seen. CONCLUSIONS: There are many advantages in HDR-B, but the most important ones are the capability of on-line dosimetry, quality control and the procedure being very conformal. There is a low incidence of GU and GI acute and late morbidity with acceptable bNED when treating initial and locally advanced prostate cancer with HDR-B as a boost to EBRT, but we still need to wait for results of phase III open trials that analyze HDR-B and conformal therapy. PMID- 12648790 TI - Pre- and post-implant dosimetry: an inter-comparison between UK prostate brachytherapy centres. AB - The consistency and reliability of five prostate brachytherapy centres performing pre- and post-implant dosimetry were evaluated. Ultrasound based pre-planning protocols and techniques were similar although they may be slightly affected by the chosen seed activity. Computerised tomography-based post-implant dosimetry varied because of differing estimations of the prostate volume. PMID- 12648791 TI - Inverse and forward optimization of one- and two-dimensional intensity-modulated radiation therapy-based treatment of concave-shaped planning target volumes: the case of prostate cancer. AB - BACKGROUND: Intensity-modulated radiation therapy (IMRT) was suggested as a suitable technique to protect the rectal wall, while maintaining a satisfactory planning target volume (PTV) irradiation in the case of high-dose radiotherapy of prostate cancer. However, up to now, few investigations tried to estimate the expected benefit with respect to conventional three-dimensional (3D) conformal radiotherapy (CRT). PURPOSE: Estimating the expected clinical gain coming from both 1D and 2D IMRT against 3DCRT, in the case of prostate cancer by mean of radiobiological models. In order to enhance the impact of IMRT, the case of concave-shaped PTV including prostate and seminal vesicles (P+SV) was considered. MATERIALS AND METHODS: Five patients with concave-shaped PTV including P+SV were selected. Two different sets of constraints were applied during planning: in the first one a quite large inhomogeneity of the dose distribution within the PTV was accepted (set (a)); in the other set (set (b)) a greater homogeneity was required. Tumor control probability (TCP) and normal tissue control probability (NTCP) indices were calculated through the Webb-Nahum and the Lyman-Kutcher models, respectively. Considering a dose interval from 64.8 to 100.8 Gy, the value giving a 5% NTCP for the rectum was found (D(NTCP(rectum)=5%)) using two different methods, and the corresponding TCP(NTCP(rectum)=5%) and NTCP(NTCP(rectum)=5%) for the other critical structures were derived. With the first method, the inverse optimization of the plans was performed just at a fixed 75.6 Gy ICRU dose; with the second method (applied to 2/5 patients) inverse treatment plannings were re-optimized at many dose levels (from 64.8 to 108 Gy with 3.6 Gy intervals). In this case, three different values of alpha/beta (10, 3, 1.5)were used for TCP calculation. The 3DCRT plan consisted of a 3-fields technique; in the IMRT plans, five equi-spaced beams were applied. The Helios Inverse Planning software from Varian was used for both the 2D IMRT and the 1D IMRT inverse optimization, the last one being performed fixing only one available pair of leaves for modulation. A previously proposed forward 1D IMRT 'class solution' technique was also considered, keeping the same irradiation geometry of the inversely optimized IMRT techniques. RESULTS: With the first method, the average gains in TCP(NTCP(rectum)=5%) of the 2D IMRT technique, with respect 3DCRT, were 10.3 and 7.8%, depending on the choice of the DVHs constraints during the inverse optimization procedure (set (a) and set (b), respectively). The average gain (DeltaTCP(NTCP(rectum)=5%)) coming from the inverse 1D IMRT optimization was 5.0%, when fixing the set (b) DVHs constraints. Concerning the forward 1D IMRT optimization, the average gain in TCP(NTCP(rectum)=5%) was 4.5%. The gain was found to be correlated with the degree of overlapping between rectum and PTV. When comparing 2D IMRT and 1D IMRT, in the case of the more realistic set (b) constraints, DeltaTCP(NTCP(rectum)=5%) was always less than 3%, excepting one patient with a very large overlap region. Basing our choice on this result, the second method was applied to this patient and one of the remaining. Through the inverse re-optimization of the treatment plans at each dose level, the gain in TCP(NTCP(rectum)=5%) of the inverse 2D technique was significantly higher than the ones obtained by applying the first method (concerning the two patients: +6.1% and +2.4%), while no significant benefit was found for inverse 1D. The impact of changing the alpha/beta ratio was less evident in the patient with the lower gain in TCP(NTCP(rectum)=5%). CONCLUSIONS: The expected benefit due to IMRT with respect to 3DCRT seems to be relevant when the overlap between PTV and rectum is high. Moreover, the difference between the inverse 2D and the simpler inverse or forward 1D IMRT techniques resulted in being relatively modest, with the exception of one patient, having a very large overlap between rectum and PTV. Optimizing the inverse planning at each dose level to find TCP(NTCP(rectum)=5%)e level to find TCP(NTCP(rectum)=5%) can improve the performances of inverse 2D IMRT, against a significant increase of the time for planning. These results suggest the importance of selecting the patients that could have significant benefit from the application of IMRT. PMID- 12648792 TI - Dosimetric implications of changes in patient repositioning and organ motion in conformal radiotherapy for prostate cancer. AB - PURPOSE: To assess the influence of patient repositioning and organ motion on dose distribution within the prostate and the seminal vesicles (clinical target volume, (CTV)). MATERIAL AND METHODS: Nine patients were simulated and treated in the supine position, with an empty bladder, and without immobilization devices. While on treatment, patients underwent weekly pelvic computed tomography (CT) scans under conditions identical to those at simulation. Patients were aligned using lasers on anterior and lateral skin tattoos, onto which lead markers were placed. After each CT scan (n=53) the CTV was redefined by contouring, and a new isocenter was obtained. A six-field technique was used. The field margins around the CTV were 20 mm in the cranio-caudal axis, and 13 mm in the other axes, except in the lateral fields where a 10 mm posterior margin was used. Dose-volume histograms (DVHs) for each organ were compared with those determined at simulation, using the notion of the proportional change in the area under the CTV DVH curve resulting from a change in treatment plan (cDVH). RESULTS: The reproducibility of the dose distribution was good for the prostate (%cDVH, mean+/ SD: -0.97+/-2.11%) and less than optimal for the seminal vesicles (%cDVH, mean+/ SD: -4.66+/-10.45%). When correlating prostate %cDVH variations with displacements of the isocenter in the Y axis (antero-posterior) the %cDVH exceeded (-)5% in only two dosimetries, both with an isocenter shift of >10 mm. For the seminal vesicles, however, ten out of 53 dosimetries showed a %cDVH exceeding (-) 5%. In nine of these ten dose distribution studies the posterior shift of the isocenter exceeded 8 mm. CONCLUSIONS: Precise targeting of prostate radiotherapy is primarily dependent on careful daily set-up and on random changes in rectal geometry. Margins no less than 10 mm around the prostate and at least 15 mm around the seminal vesicles are probably necessary to insure adequate target coverage with a six-field technique. PMID- 12648793 TI - Radiotherapy treatment planning of prostate cancer using magnetic resonance imaging alone. AB - PURPOSE: Accurate anatomical delineation of the gross tumour volume (GTV) is crucial for effective radiotherapy (RT) treatment of prostate cancers. Although reference to pelvic magnetic resonance (MR) for improved delineation of the prostate is a regular practice in some clinics, MR has not replaced CT due to its geometrical distortions and lack of electron-density information. The possibility and practicality of using MR only for RT treatment planning were studied. MATERIALS AND METHODS: The addition of electron-density information to MR images for conformal radiotherapy (CRT) planning of the prostate was quantified by comparing dose distributions created on the homogeneous density- and bulk-density assigned images to original CT for four patients. To quantify the MR geometrical distortions measurements of a phantom imaged in CT (Siemens Somatom Plus 4) and FLASH 3D T1-weighted MR (1.5 T whole body Siemens Magnetom Vision) were compared. Dose statistics from CRT treatment plans made on CT and MR for five patient data were compared to determine if MR-only treatment plans can be made. RESULTS: The differences between dose-plans on bulk-density assigned images when compared to CT were less than 2% when water and bone values were assigned. Dose differences greater than 2% were observed when images of homogeneous-density assignment were compared to the CT. Phantom measurements showed that the distortions in the FLASH 3D T1-weighted MR averaged 2 mm in the volume of interest for prostate RT planning. For the CT and MR prostate planning study, doses delivered to the planning target volume (PTV) in CT and MR were always inside a 93-107% dose range normalised to the isocentre. Also, the doses to the organs-at-risk in the MR images were similar to the doses delivered to the volumes in the registered CT image when the organ volumes between the two images were similar. CONCLUSIONS: Negligible differences were observed in dose distribution between CRT plans using bone+water CT number bulk-assigned image and original CT. Also, the MR distortions were reduced to negligible amounts using large bandwidth MR sequence for prostate CRT planning. MR treatment planning was demonstrated using a large bandwidth sequence and bulk-assigned images. The development of higher quality, low distortion MR sequence will allow regular practice of this technique. PMID- 12648794 TI - Magnetic resonance imaging in the radiation treatment planning of localized prostate cancer using intra-prostatic fiducial markers for computed tomography co registration. AB - PURPOSE: To assess the feasibility, and potential implications, of using intra prostatic fiducial markers, rather than bony landmarks, for the co-registration of computed tomography (CT) and magnetic resonance (MR) images in the radiation treatment planning of localized prostate cancer. METHODS: All men treated with conformal therapy for localized prostate cancer underwent routine pre-treatment insertion of prostatic fiducial markers to assist with gross target volume (GTV) delineation and to identify prostate positioning during therapy. Six of these men were selected for investigation. Phantom MRI measurements were obtained to quantify image distortion, to determine the most suitable gold alloy marker composition, and to identify the spin-echo sequences that optimized both marker identification and the contrast between the prostate and the surrounding tissues. The GTV for each patient was contoured independently by three radiation oncologists on axial planning CT slices, and on axial MRI slices fused to the CT slices by matching the implanted fiducial markers. From each set of contours the scan common volume (SCV), and the scan encompassing volume (SEV), were obtained. The ratio SEV/SCV for a given scan is a measure of inter-observer variation in contouring. For each of the 18 patient-observer combinations the observer common volume (OCV) and the observer encompassing volume (OEV) was obtained. The ratio OEV/OCV for a given patient-observer combination is a measure of the inter modality variation in contouring. The distance from the treatment planning isocenter to the prostate contours was measured and the discrepancy between the CT- and the MR-defined contour recorded. The discrepancies between the CT- and MR defined contours of the posterior prostate were recorded in the sagittal plane at 1-cm intervals above and below the isocenter. RESULTS: Phantom measurements demonstrated trivial image distortion within the required field of view, and an 18K Au/Cu alloy to be the marker composition most suitable for CT-MRI image fusion purposes. Inter-observer variation in prostate contouring was significantly less for MR compared to CT. The mean SEV/SCV ratio was 1.58 (confidence interval (CI): 1.47-1.69) for CT scans and 1.37 (CI: 1.33-1.41) for MR scans (paired t-test; P=0.036). The overall magnitude of contoured GTV was similar for MR and CT; however, there were spatial discrepancies in contouring between the two modalities. The greatest systematic discrepancy was at the posterior apical prostate border, which was defined 3.6 mm (SD 3.5 mm) more posterior on MR- than CT-defined contouring. CONCLUSIONS: Prostate contouring on MR is associated with less inter-observer variation than on CT. In addition, we have demonstrated the feasibility of using intra-prostatic fiducial markers, rather than bony landmarks, for the co-registration of CT and MR images in the radiation treatment planning of localized prostate cancer. This technique, together with on-line correction of treatment set-up according to the fiducial marker position on electronic portal imaging, may enable a reduction in the planning target volume (PTV) margin needed to account for inter-observer error in target delineation, and for prostate motion. PMID- 12648795 TI - Feasibility of geometrical verification of patient set-up using body contours and computed tomography data. AB - BACKGROUND AND PURPOSE: Body contours can potentially be used for patient set-up verification in external-beam radiotherapy and might enable more accurate set-up of patients prior to irradiation. The aim of this study is to test the feasibility of patient set-up verification using a body contour scanner. MATERIAL AND METHODS: Body contour scans of 33 lung cancer and 21 head-and-neck cancer patients were acquired on a simulator. We assume that this dataset is representative for the patient set-up on an accelerator. Shortly before acquisition of the body contour scan, a pair of orthogonal simulator images was taken as a reference. Both the body contour scan and the simulator images were matched in 3D to the planning computed tomography scan. Movement of skin with respect to bone was quantified based on an analysis of variance method. RESULTS: Set-up errors determined with body-contours agreed reasonably well with those determined with simulator images. For the lung cancer patients, the average set up errors (mm)+/-1 standard deviation (SD) for the left-right, cranio-caudal and anterior-posterior directions were 1.2+/-2.9, -0.8+/-5.0 and -2.3+/-3.1 using body contours, compared to -0.8+/-3.2, -1.0+/-4.1 and -1.2+/-2.4 using simulator images. For the head-and-neck cancer patients, the set-up errors were 0.5+/-1.8, 0.5+/-2.7 and -2.2+/-1.8 using body contours compared to -0.4+/-1.2, 0.1+/-2.1, 0.1+/-1.8 using simulator images. The SD of the set-up errors obtained from analysis of the body contours were not significantly different from those obtained from analysis of the simulator images. Movement of the skin with respect to bone (1 SD) was estimated at 2.3 mm for lung cancer patients and 1.7 mm for head-and-neck cancer patients. CONCLUSION: Measurement of patient set-up using a body-contouring device is possible. The accuracy, however, is limited by the movement of the skin with respect to the bone. In situations where the error in the patient set-up is relatively large, it is possible to reduce these errors using a computer-aided set-up technique based on contour information. PMID- 12648796 TI - Thermoluminescent dosimeters (TLD) quality assurance network in the Czech Republic. AB - INTRODUCTION: The Czech thermoluminescent dosimeters (TLD) quality assurance network was established in 1997. Its aim is to pursue a regular independent quality audit in Czech radiotherapy centres and to support state supervision. MATERIALS AND METHODS: The audit is realised via mailed TL dosimetry. The TLD system consists of encapsulated LiF:Mg,Ti powder (type MT-N) read with Harshaw manual reader model 4000. Basic mode of the TLD audit covers measurements under reference conditions, specifically beam calibration checks for all clinically used photon and electron beams. Advanced mode consists of measurements under both reference and non-reference conditions using a solid multipurpose phantom ('Leuven phantom') for photon beams. The radiotherapy centres are instructed to deliver to the TLD on central beam axis absorbed dose of 2 Gy calculated with their treatment planning system for a particular treatment set-up. The TLD measured doses are compared with the calculated ones. Deviations of +/-3% are considered acceptable for both basic and advanced mode of the audit. RESULTS: There are 34 radiotherapy centres in the Czech Republic. They undergo the basic mode of the TLD audit regularly every 2 years. If a centre shows a deviation outside the acceptance level, it is audited more often. Presently, most of the checked beams comply with the acceptance level. The advanced TLD audit has been implemented as a pilot study for the present. The results were mostly within the acceptance limit for the measurements on-axis, whereas for off-axis points they fell beyond the limit more frequently, especially for set-ups with inhomogeneities, oblique incidence and wedges. CONCLUSIONS: The results prove the importance of the national TLD quality assurance network. It has contributed to the improvement of clinical dosimetry in the Czech Republic. In addition, it helps the regulatory authority to monitor effectively and regularly radiotherapy centres. PMID- 12648798 TI - Implications of SENSE MR in routine clinical practice. AB - Sensitivity encoding (SENSE) uses multiple MRI receive coil elements to encode spatial information in addition to traditional gradient encoding. Requiring less gradient encodings translates into shorter scan times, which is extremely beneficial in many clinical applications. SENSE is available to routine diagnostic imaging for the past 2 years. This paper highlights the use of SENSE with scan time reduction factors up to 6 in contrast-enhanced MRA, routine abdominal imaging, mammography, cardiac and neuro imaging. It is shown that SENSE has opened new horizons in both routine and advanced MR imaging. PMID- 12648799 TI - TrueFISP--technical considerations and cardiovascular applications. AB - Magnetic resonance imaging (MRI) using steady-state free precession (SSFP) sequences was described in the early years of MR imaging. However, due to hardware imperfections, these techniques were not robust enough at the time to play any significant role in clinical MRI. More recently, significant hardware improvements became widely available, and the SSFP sequences such as TrueFISP (true fast imaging with steady-state precession) became very popular for a variety of clinical applications due to the distinct improvements in signal-to noise ratio (SNR) and contrast-to-noise ratio (CNR). The purpose of this article is to give an overview on the basics of TrueFISP imaging and to demonstrate its potential for current clinical applications with a focus on cardiovascular MRI. PMID- 12648800 TI - Tissue-specific MR contrast agents. AB - The purpose of this review is to outline recent trends in contrast agent development for magnetic resonance imaging. Up to now, small molecular weight gadolinium chelates are the workhorse in contrast enhanced MRI. These first generation MR contrast agents distribute into the intravascular and interstitial space, thus allowing the evaluation of physiological parameters, such as the status or existence of the blood-brain-barrier or the renal function. Shortly after the first clinical use of paramagnetic metallochelates in 1983, compounds were suggested for liver imaging and enhancing a cardiac infarct. Meanwhile, liver specific contrast agents based on gadolinium, manganese or iron become reality. Dedicated blood pool agents will be available within the next years. These gadolinium or iron agents will be beneficial for longer lasting MRA procedures, such as cardiac imaging. Contrast enhanced lymphography after interstitial or intravenous injection will be another major step forward in diagnostic imaging. Metastatic involvement will be seen either after the injection of ultrasmall superparamagnetic iron oxides or dedicated gadolinium chelates. The accumulation of both compound classes is triggered by an uptake into macrophages. It is likely that similar agents will augment MRI of atheriosclerotic plaques, a systemic inflammatory disease of the arterial wall. Thrombus-specific agents based on small gadolinium labeled peptides are on the horizon. It is very obvious that the future of cardiovascular MRI will benefit from the development of new paramagnetic and superparamagnetic substances. The expectations for new tumor-, pathology- or receptor-specific agents are high. However, is not likely that such a compound will be available for daily routine MRI within the next decade. PMID- 12648801 TI - MR imaging at high magnetic fields. AB - Recently, more investigators have been applying higher magnetic field strengths (3-4 Tesla) in research and clinical settings. Higher magnetic field strength is expected to afford higher spatial resolution and/or a decrease in the length of total scan time due to its higher signal intensity. Besides MR signal intensity, however, there are several factors which are magnetic field dependent, thus the same set of imaging parameters at lower magnetic field strengths would provide differences in signal or contrast to noise ratios at 3 T or higher. Therefore, an outcome of the combined effect of all these factors should be considered to estimate the change in usefulness at different magnetic fields. The objective of this article is to illustrate the practical scientific applications, focusing on MR imaging, of higher magnetic field strength. First, we will discuss previous literature and our experiments to demonstrate several changes that lead to a number of practical applications in MR imaging, e.g. in relaxation times, effects of contrast agent, design of RF coils, maintaining a safety profile and in switching magnetic field strength. Second, we discuss what will be required to gain the maximum benefit of high magnetic field when the current magnetic field (< or = 1.5 T) is switched to 3 or 4 T. In addition, we discuss MR microscopy, which is one of the anticipated applications of high magnetic field strength to understand the quantitative estimation of the gain benefit and other considerations to help establish a practically available imaging protocol. PMID- 12648802 TI - MR diffusion tensor imaging: recent advance and new techniques for diffusion tensor visualization. AB - Recently, diffusion tensor imaging is attracting the biomedical researchers for its application in depiction of fiber tracts based on diffusion anisotropy. In this paper, we briefly describe the basic theory of diffusion tensor MR imaging, the determination process of diffusion tensor, and the basic concepts of diffusion tensor visualization techniques. Several results of clinical application in our institute are also introduced. Finally, the limitations, advantages and disadvantages of the techniques are discussed for further application of diffusion tensor visualization. PMID- 12648803 TI - MR imaging of ischemic penumbra. AB - Cerebral ischemic stroke is one of the most fatal diseases despite current advances in medical science. Recent demonstration of efficacy using intravenous and intra-arterial thrombolysis demands therapeutic intervention tailored to the physiologic state of the individual tissue and stratification of patients according to the potential risks for therapies. In such an era, the role of the neuroimaging becomes increasingly important to evaluate the extent and location of tissues at risk of infarction (ischemic penumbra), to distinguish it from unsalvageable infarcted tissues or doomed hemorrhagic parenchyma. In this review, we present briefly the current role and limitation of computed tomography and conventional magnetic resonance imaging (MRI). We also present the possible applications of advanced MR techniques, such as diffusion and perfusion imaging, concentrating on the delineation or detection of ischemic penumbra. PMID- 12648804 TI - Anti-nociceptive and anti-inflammatory effects of methanolic extract of Asparagus pubescens root in rodents. AB - The effect of methanolic extract of Asparagus pubescens was investigated on chemical, thermal-induced pain as well as fresh egg albumin-induced inflammation and pentylenetetrazol (PTZ)-induced convulsion in rodents. The extract dose dependently (0.25-1.5 g/kg) inhibited acetic acid-induced writhing, formalin induced pain licking and hot plate-induced pain in mice. The extract significantly inhibited both the fresh egg albumin-induced inflammation in rats as well as PTZ-induced convulsion in mice. These inhibitions were statistically significant (P < 0.02-0.001). It increased the latencies of both clonic and tonic convulsions and delayed their mortalities. Its ability to reduce both neurogenic and non-neurogenic pains may be related to its active constituents such as tannins, saponins, steroid and flavonoids. PMID- 12648805 TI - Screening of antioxidant activity of three Indian medicinal plants, traditionally used for the management of neurodegenerative diseases. AB - A number of Indian medicinal plants have been used for thousands of years in the traditional system of medicine (Ayurveda). Amongst these are plants used for the management of neurodegenerative diseases such as Parkinson's, Alzheimer's, loss of memory, degeneration of nerves and other neuronal disorders by the Ayurvedic practitioners. Though the etiology of neurodegenerative diseases remains enigmatic, there is evidence, which indicates that defective energy metabolism, excitotoxicity and oxidative damage may be crucial factors (Ann. Neurol. 38 (3) (1995) 357). The part of the Ayurvedic system that provides an approach to prevention and treatment of degenerative diseases is known as Rasayana, and plants used for this purpose are classed as rejuvenators. This group of plants generally possesses strong antioxidant activity (Pharmacol. Biochem. Behav. 43 (1992) 1175), but only a few have been investigated in detail. In the present study, three such rasayana plants were tested for the first time for their toxicity and free radical scavenging activity both in vitro and ex vivo. All the three plant infusions (up to 1 mg/ml) showed no toxic effects on the viability of PC12 cell line as judged by MTT-test. Both ethanolic extracts and water infusions of the plants were tested for their antioxidant activity in the 2,2'-azinobis-3 ethyl-benzothiazoline-6-sulfonic acid (ABTS*(+)) radical cation decolorization assay; inhibition of lipid peroxidation by plant infusions was carried out using spontaneous lipid peroxidation of rat brain homogenate, and IC50 values were determined. The results from the ABTS assay showed that the ethanolic extract of Sida cordifolia was found to be most potent (IC50 16.07 microg/ml), followed by Evolvulus alsinoides (IC50 33.39 microg/ml) and Cynodon dactylon (IC50 78.62 microg/ml). The relative antioxidant capacity for the water infusions was observed in the following order: E. alsinoides (IC50 172.25 microg/ml)>C. dactylon (IC50 273.64 microg/ml)>S. cordifolia (IC50 342.82 microg/ml). The results of water infusions of the plants on lipid peroxidation were as follows: E. alsinoides (IC50 89.23 microg/ml)>S. cordifolia) (IC50 126.78 microg/ml)>C. dactylon (IC50 608.31 microg/ml). PMID- 12648806 TI - Hypoglycemic properties of aqueous bark extract of Ceiba pentandra in streptozotocin-induced diabetic rats. AB - The claim by Nigerian traditional herbal medicine practitioners that the silk cotton tree, Ceiba pentandra L. Gaertn (Bombacaceae) barks extract has antidiabetic properties was investigated. Diabetes mellitus was induced with streptozotocin and graded doses of the aqueous bark extract were then administered ad libitum in drinking water to the experimentally diabetic rats for 28 days. Administration of the aqueous bark extract caused a statistically significant reduction in plasma glucose level in streptozotocin induced diabetic rats. The extract appeared non-toxic as evidenced by normal serum levels of AST, ALT, ALP and bilirubin. The data appear to support the hypoglycemic effects of C. pentandra. PMID- 12648807 TI - Protective effect of Rhodiola sachalinensis extract on carbon tetrachloride induced liver injury in rats. AB - This study was carried out to investigate the protective effect of an aqueous extract from the root of Rhodiola sachalinensis (RSE) on liver injury induced by repetitive administration of carbon tetrachloride in rats. RSE was given orally to rats at doses of 50, 100 or 200 mg/kg throughout the carbon tetrachloride treatment for 28 days. In rats treated with carbon tetrachloride, the levels of hydroxyproline and malondialdehyde (MDA) in the liver, and serum enzyme activities were significantly increased. RSE treatment significantly reduced the levels of liver hydroxyproline and MDA, and serum enzyme activities, in accordance with improved histological findings. Immunohistological findings indicated RSE treatment inhibited hepatic stellate cell activation, which is a major step for collagen accumulation during liver injury. These data suggest that RSE protects the liver from repetitive injury induced by carbon tetrachloride in rats. PMID- 12648808 TI - Biochemical studies on the anti-ulcerogenic potential of Hemidesmus indicus R.Br. var. indicus. AB - Roots of Hemidesmus indicus var. indicus are used for various ailments in Indian traditional medicines. The present study evaluated the antiulcerogenic property of aqueous ethanolic extracts of the roots in animal models. Modified pyloric ligated (Shay) rat model and aspirin-induced ulcerogenesis in pylorus ligated rat models were used for this study and analysed for gastric volume, ulcer score, pH, free and total acidity and sodium and potassium ion output. Bio-chemical estimations like total proteins, total hexoses, hexosamine, fucose, sialic acid and pepsin were also made. Ulcer score was calculated for cysteamine-induced duodenal ulcer model. Roots collected during flowering season were found to be more effective than that collected during vegetative seasons. PMID- 12648809 TI - Cytotoxic and antitumor activities of a polypore macrofungus, Phellinus rimosus (Berk) Pilat. AB - Cytotoxic and antitumor activities of ethyl acetate, methanol and aqueous extracts of a wood inhabiting polypore macrofungus, Phellinus rimosus (Berk) Pilat. were studied. Ethyl acetate and methanol extracts showed in vitro cytotoxic activity against Dalton's lymphoma ascites (DLA) and Ehrlich's ascites carcinoma (EAC) cell lines. The aqueous extract did not exhibit cytotoxicity against the tested cell lines. All the three extracts were highly effective in inhibiting growth of solid tumor induced by DLA cell line in mice. However, the antitumor activity of ethyl acetate extract was higher than that of methanol and aqueous extracts. The ethyl acetate extract was also effective in preventing the EAC induced ascites tumor development in mice. The antitumor activity of all the three extracts against solid tumor at a dose of 50 mg/kg (p.o.) was comparable to the clinically used standard reference drug, cisplatin (4 mg/kg, i.p.). Pre treatment of the extracts was also effective in inhibiting the tumor growth induced by DLA cell lines. The experimental results revealed that ethyl acetate extract of P. rimosus possessed significant antitumor activity. The findings thus suggest the potential use of this mushroom as antitumor agent. PMID- 12648810 TI - Effect of Coccinia indica leaves on antioxidant status in streptozotocin-induced diabetic rats. AB - The antioxidant effect of an ethanolic extract of Coccinia indica leaves, an indigenous plant used in Ayurvedic medicine in India, was studied in Streptozotocin-diabetic rats. Oral administration of Coccinia indica leaf extract (CLEt) (200 mg/kg body weight) for 45 days resulted in a significant reduction in thiobarbituric acid reactive substances and hydroperoxides. The extract also causes a significant increase in reduced glutathione, superoxide dismutase, catalase, glutathione peroxidase and glutathione-S-transferase in liver and kidney of streptozotocin diabetic rats, which clearly shows the antioxidant property of CLEt. The effect of CLEt at 200 mg/kg body weight was more effective than glibenclamide PMID- 12648811 TI - Anti-inflammatory and anti-nociceptive effects of Sphaeranthus senegalensis. AB - Sphaeranthus senegalensis Vaill (Asteraceae) is used in traditional medicine as a remedy for rheumatic pains and other ailments. The anti-inflammatory and anti nociceptive activity of the aqueous extract of the whole shoot of the plant was evaluated in mice and rats. Activity of the extract against egg-albumin induced hind paw oedema was measured to evaluate the anti-inflammatory activity while the anti-nociceptive potency was evaluated using three painful stimuli viz acetic acid induced abdominal constriction and hot plate test in mice, and the formalin test in rats. Results indicated that the extract possess remarkable dose dependent anti-inflammatory activities in rats. The extract also showed anti nociceptive activities against acetic acid induced writhing, formalin and the hot plate pain models. The effects were significant (P < 0.05) when compared with the saline control group. The results suggest the presence of a potent anti inflammatory and anti-nociceptive principle in the extract, which support the folkloric use of the plant in relieving rheumatic pains. PMID- 12648812 TI - Antispasmodic effect of Achillea nobilis L. subsp. sipylea (O. Schwarz) Bassler on the rat isolated duodenum. AB - The aim of the present study was to investigate the antispasmodic effect of the total extract of Achillea nobilis L. subsp. sipylea (Schwarz) Bassler (Asteraceae) on rat duodenum. In the first part of experiments, cumulative dose response curves for acetylcholine (Ach) were obtained and then dose-response curves are repeated after addition of atropine, papaverine and different doses of the extract. In the second part, cumulative dose-response curves to CaCl(2) were obtained in the absence and presence of verapamil and different doses of the extract. In the third part, papaverine and extract were applied to the tissues after contraction with K(+). The extract has exhibited an inhibitory effect on the dose-response curves induced by Ach and CaCl(2) on rat duodenum and significantly reduced the maximal response in a concentration-dependent manner. A similar effect was observed with papaverine but not with atropine on the dose response curves obtained by ACh. Verapamil also reduced the maximal response in curves induced by CaCl(2). The present results demonstrate that total extract of A. nobilis subsp. sipylea exerts antispasmodic activity on rat duodenum. PMID- 12648813 TI - Kuwanon G: an antibacterial agent from the root bark of Morus alba against oral pathogens. AB - Kuwanon G was isolated from the ethyl acetate fraction of methanol extract of Morus alba and its structure was elucidated by 13C-NMR, 1H-NMR and FAB-MS. Antibacterial activity of kuwanon G was investigated by the minimum inhibitory concentration (MIC) test and the viable cell count method. MIC of kuwanon G against Streptococcus mutans causing dental caries was determined to be 8.0 microg/ml. The bactericidal test showed that kuwanon G completely inactivated S. mutans at the concentration 20 microg/ml in 1 min. Kuwanon G also significantly inhibited the growth of other cariogenic bacteria such as Streptococcus sobrinus and Streptococcus sanguis, and Porpyromonas gingivalis causing periodontitis. Transmission electron microscopy (TEM) of kuwanon G treated cells demonstrated remarkable morphological damage of the cell wall and condensation of the cytoplasm. PMID- 12648814 TI - Pharmacokinetics and bioavailability of ginsenoside Rb1 and Rg1 from Panax notoginseng in rats. AB - Panax notoginseng is used as a therapeutic agent to stop haemorrhages and a tonic to promote health in Chinese medicine. Currently saponins of P. notoginseng (PNS) are especially given attentions for their hemorheological properties. The pharmacokinetic profiles of the main PNS are still not accurately investigated. Therefore, our preliminary aim is to elucidate the pharmacokinetic features of ginsenoside Rb(1) (Rb(1)) and ginsenoside Rg(1) (Rg(1)), two of the main PNS in rats. Firstly, quantitive analysis of Rb(1) and Rg(1) in saponins of P. notoginseng was studied and the most suitable assay method by HPLC for blood sample were established. Then Rb(1) and Rg(1) in the same serum were determined after administering PNS to rats. The decline of Rb(1) in serum could be described by a two-compartment model. The half-life of alpha phase was 23.40 min and that of beta phase was 17.96 h. Rb(1) was absorbed from the digestive tract and the bioavailability via P.O. was 4.35%. The pharmacokinetics of Rg(1) in rats also could be described by a two-compartment model. The half-lives of Rg(1) were 24.23 min for alpha phase and 14.13 h for beta phase. Rg(1) could be absorbed in the digestive tract and the oral bioavailability was 18.40%. Both of the low oral bioavailability of Rb(1) and rapid reduction of Rg(1) in blood indicated that formula modification is necessary. PMID- 12648815 TI - Immunomodulatory effects of aqueous-extracted Astragali radix in methotrexate treated mouse spleen cells. AB - The present study was conducted to evaluate the immunomodulatory effect of aqueous-extracted Astragali radix (ARE) in methotrexate (MTX)-treated mouse spleen cells. In spleen cell proliferation assay, ARE enhanced mitogenic activity in the dose-response manner. We also investigated the effect of ARE on the reducing of immune suppression caused by MTX in mouse spleen cells. MTX decreased the spleen cell proliferation (IC(50):800 microg/ml). However, ARE significantly reduced the suppression of cell proliferation by MTX in mouse spleen cells. Immunomodulatory effect of ARE were further investigated using reverse transcription polymerase chain reaction (RT-PCR). In RT-PCR, we examined the expressions of various cytokines such as IL-6, IL-1alpha, IL-1beta, IL-12p40, GM CSF and TNF. Enhancement of IL-1alpha and IL-12p40 mRNA expressions were shown in mouse spleen cells by ARE. In spite of MTX treatment, the expressions of IL 1alpha and IL-12p40 mRNA sustained in spleen cells. These data indicate that (1) ARE has a protective effect of immune suppression, and (2) the immunomodulatory effects of ARE may be, in part, associated with the expressions of IL-1alpha and IL-12p40 mRNA as well as the mitogenic effect on spleen cells. PMID- 12648816 TI - Effects of Crocus sativus petals' extract on rat blood pressure and on responses induced by electrical field stimulation in the rat isolated vas deferens and guinea-pig ileum. AB - We have investigated the effects of Crocus sativus petals' extract on blood pressure in anaesthetised rats and also on responses of the isolated rat vas deferens and guinea-pig ileum induced by electrical field stimulation (EFS). Aqueous and ethanol extracts of C. sativus petals reduced the blood pressure in a dose-dependent manner. For example administration of 50 mg/100 g of aqueous extract changed the blood pressure from 133.5+/-3.9 to 117+/-2.1 (mmHg). EFS of the isolated rat vas deferens and guinea-pig ileum evoked contractions were decreased by aqueous and ethanol extracts of C. sativus petals. The aqueous extract (560 mg/ml) significantly reduced the contractile responses of vas deferens to epinephrine (1 microM) without any change in contraction induced by KCl (300 mM). The present results may suggest that the relaxatory action of C. sativus petals' extract on contraction induced by EFS in the rat isolated vas deferens is a postsynaptic effect. PMID- 12648817 TI - Syzigium cumini seed extracts reduce tissue damage in diabetic rat brain. AB - Syzigium cumini commonly known as Jamun, is widely used in different parts of India for the treatment of diabetes mellitus. Oral administration of an aqueous Jamun seed extract (JSEt) for 6 weeks caused a significant decrease in lipids, thiobarbituric acid reactive substances (TBARS) and an increase in catalase and superoxide dismutase in the brain of alloxan induced diabetic rats. Oral administration of an alcoholic JSEt for 6 weeks brought back all the parameters to near normal. The effect of alcoholic JSEt (100 mg/kg) was better than aqueous JSEt (5 g/kg). The effect of both these extracts was better than glibenclamide (600 microg/kg). Thus, our study shows that S. cumini seed extracts reduce tissue damage in diabetic rat brain. PMID- 12648818 TI - Studies on pharmaceutical ethnobotany in the region of Pallars (Pyrenees, Catalonia, Iberian Peninsula). Part II. New or very rare uses of previously known medicinal plants. AB - An ethnobotanical survey was carried out in the region called Pallars--consisting of two districts, Pallars Sobira and Pallars Jussa--, situated in the Central Pyrenees, in North West Catalonia (Iberian Peninsula), with an approximate area of 2530 km(2) and a population of 19,000. Through interviews with 264 people, we obtained data on 437 plant species used for health care. We detected 867 unreported or uncommon uses corresponding to 272 plant species, 52 of which had never or very rarely been cited as medicinal. This is the second paper of a series intended to present the most important findings concerning the ethnopharmacology of the area studied; it includes unreported or very uncommon uses of known medicinal plant species. A previous paper reported the general results and the new or very scarcely reported medicinal vascular plants, and a further one will deal with medicinal non vascular plants. PMID- 12648819 TI - Studies on pharmaceutical ethnobotany in the region of Pallars (Pyrenees, Catalonia, Iberian Peninsula). Part III. Medicinal uses of non-vascular plants. AB - An ethnobotanical survey was carried out in the region called Pallars- constituted by two districts, Pallars Sobira and Pallars Jussa--, situated in the Central Pyrenees, in North West Catalonia (Iberian Peninsula), with an approximate area of 2530 km(2) and a population of 19000. Through interviews with 264 people, we obtained data on 437 plant species used for health care. We detected 867 unreported or uncommon uses corresponding to 272 plant species, 52 of which had never or very rarely been cited as medicinal. This is the third paper of a series intended to present the most important findings concerning the ethnopharmacology of the area studied; it includes information on 18 pharmaceutical uses of 16 non-vascular plant species (two algae, eight fungi, five lichens and one moss). Two previous papers reported the general results and the new or very scarcely reported medicinal vascular plants, and the unreported or very uncommon uses of already known medicinal plants. PMID- 12648820 TI - In vitro antiplasmodial activity of some plants used in Kisii, Kenya against malaria and their chloroquine potentiation effects. AB - Fifty-five organic and aqueous extracts of 11 plants used in malaria therapy in Kisii District, Kenya were tested in vitro against chloroquine (CQ)-sensitive and resistant strains of Plasmodium falciparum. Of the plants tested, 73% were active (IC(50) < 100 microg/ml). Three plants, Vernonia lasiopus, Rhamnus prinoides and Ficus sur afforded extracts with IC(50) values ranging less than 30 microg/ml against both CQ-sensitive and resistant strains. Combination of some extracts with CQ against the multi-drug resistant P. falciparum isolate V1/S revealed some synergistic effect. The plant extracts with low IC(50) values may be used as sources for novel antimalarial compounds to be used alone or in combination with CQ. PMID- 12648821 TI - Hypoglycemic activity of Urtica pilulifera in streptozotocin-diabetic rats. AB - The hypoglycemic activity of lectin isolated from Urtica pilulifera L. seeds (Urticaceae) was investigated in streptozotocin (STZ) induced diabetic rats. Significant hypoglycemic effect was found at the dose of 100 mg/kg after i.p. administration for 30 days. Blood glucose (BG) level, food and fluid intake, body weight (BW) loss and histopathologic findings of the normal and diabetic animals were evaluated. The group treated with UPSL (U. pilulifera seed lectin) was also compared against glipizide (oral antidiabetic agent, Carlo-Erba) as a standard. PMID- 12648822 TI - Antihyperglycemic and hypoglycemic effect of Aporosa lindleyana in normal and alloxan induced diabetic rats. AB - The hypoglycemic effect of aqueous and alcoholic extracts of root of Aporosa lindleyana was investigated in both normal and alloxan induced diabetic rats. The blood glucose levels were measured at 0, 1, 2 and 3 h after the treatment. The aqueous and alcoholic extracts of A. lindleyana (100 mg/kg) reduced the blood glucose of normal rat from 80.4+/-2.7 to 69.8+/-2.0 mg% and 82.6+/-1.9 to 70.8+/ 3.2 mg%, respectively 3 h after oral administration of the extract (P < 0.001) and also significantly lowered blood glucose level in alloxan induced diabetic rat from 306+/-3.37 to 160+/-2.46 and 328+/-4.15 to 152+/-3.86 mg%, respectively 3 h after oral administration of the extract (P < 0.001). The antihyperglycemic activity of A. lindleyana was compared with tolbutamide, an oral hypoglycemic agent. PMID- 12648823 TI - Gastroprotective activity of Nigella sativa oil and its constituent, thymoquinone, against gastric mucosal injury induced by ischaemia/reperfusion in rats. AB - Ischaemia/reperfusion (I/R) induced gastric lesion, is known to be linked with free radical (FR) formation. Therefore, this model was used to assess the antioxidant effects of Nigella sativa oil (N.O) and thymoquinone (TQ) on gastric mucosal redox state and gastric lesions, 1 and 24 h after reperfusion. Male Wistar rats were subjected to I/R and were injected with either N.O (2.5 and 5 ml/kg, p.o) or TQ (5, 20, 50 and 100 mg/kg, p.o). The results showed that I/R elevated the levels of lipid peroxide (LPX) and lactate dehydrogenase (LDH), while decreased those of reduced glutathione (GSH) and superoxide dismutase (SOD). These biochemical changes were accompanied by an increase in the formation of gastric lesions, which was reduced by either treatment. N.O tended to normalize the level of LDH, GSH and SOD. However, its effect to restore LPX was only seen 24 h after reperfusion. Moreover, the aforementioned parameters were nearly reinstated by TQ. On the other hand, high doses of TQ (50 and 100 mg/kg) severely reduced the GSH content, 1 h after reperfusion. These results indicate that both N.O and TQ possess gastroprotective effect against gastric lesions which may be related to the conservation of the gastric mucosal redox state. PMID- 12648824 TI - Prevention of bone loss in calcium deficient ovariectonized rats by OST-6, a herbal preparation. AB - OST-6, a herbomineral preparation, was studied for its inhibitory effects on the progress of bone loss induced by ovariectomy (OVX) and concurrent calcium deficiency in rats. Calcium deficient ovariectomized rats were administered with OST-6 at 250 mg/kg b. w. twice a day orally for 16 weeks. Compared with sham operated animals, OVX animals showed an increase in serum ALP, urinary excretion of calcium and phosphorus, which were significantly prevented in OST-6 administered rats. Evaluation of cortical bone morphometric indices by CT Scanning technique showed an increased medullary width and cross-sectional medullary area (MA), decreased periosteal area (PA), combined cortical thickness (CCT), cortical area/periosteal area (CA/PA) and maximal metaphyseal radial density (MMRD) in OVX animals when compared with sham operated. OST-6 treatment significantly prevented these bone resorption variables. Scanning Electron Microscopy (SEM) study revealed porous and erosive appearance of femur bone at the epiphyseal region and decreased calcium to phosphorus ratio (Ca:P) in the OVX rats when compared with sham operated rats. The treatment with OST-6 prevented the epiphyseal bone resorption and maintained Ca:P ratio. The results of ash analysis indicated a reduced bone mineral content (calcium and phosphorus) and ash weight and percent ash in OVX animals when compared with sham operated animals. All results are statistically significant at P < 0.05. These finding suggest the usefulness of OST-6 in the prevention of bone loss in a natural way through utilization of herbal resources. PMID- 12648826 TI - Muscle relaxant activity of Elaeagnus angustifolia L. fruit seeds in mice. AB - Muscle relaxant effect of Elaeagnus angustifolia L. (Elaeagnaceae) fruit seeds was studied in mice using traction test. The aqueous and ethanolic extracts (i.p) induced a muscle relaxant effect in a dose dependent manner as effective as diazepam (1 mg/kg). The aqueous extract was partitioned with methanol-chloroform (MeCh) and n-butanol (Bu.) saturated with water. The MeCh and Bu. fractions did not show activity. Preliminary phytochemical tests showed that the extract contains flavonoid. The results suggested that E. angustifolia fruit seeds exerted muscle relaxant activity via flavonoid component(s). PMID- 12648825 TI - Immunomodulation by water-soluble derivative of propolis: a factor of antitumor reactivity. AB - The antimetastatic efficacy of a water-soluble derivative of propolis (WSDP) was studied. Tumor was a transplantable mammary carcinoma of CBA mouse. Metastases in the lung were generated by 2 x 10(5) viable tumor cells i.v. WSDP was given intraperitoneally at doses of 50 or 150 mg/kg before or after tumor cell inoculation. Therapies reduced the number of metastases in the lung and tumor growth was suppressed significantly by WSDP. It is likely that antimetastatic activity of the WSDP is mainly mediated by immunomodulatory activity. Changes in several immunological parameters such as production of lymphocyte activating factor by peritoneal macrophages and the efficacy of those macrophages to kill tumor cell in vitro, responses of lymphocytes to mitogen, and weight and cellularity of spleen, respectively, correlated well with antimetastatic properties of the WSDP. Based on results we postulate that the antimetastatic activity of propolis includes a pronounced immunomodulatory activity mainly toward augmentation of nonspecific antitumor resistance in mice via macrophage activation. PMID- 12648827 TI - The malian medicinal plant Trichilia emetica; studies on polysaccharides with complement fixing ability. AB - Trichilia emetica is a tree or bush that has many different traditional uses in Mali, amongst others the leaves have been used as a woundhealing remedy. As polysaccharides have been isolated from other plants that have a long tradition as woundhealing remedies, it was of interest to study the polysaccharides of T. emetica as well. The polysaccharides were extracted at 50 and 100 degrees C, and the polymeric material was separated into neutral and acidic polymers by anion exchange chromatography. The complement fixation ability of the different fractions was determined. The Acidic fraction 4 of the 100 degrees C extract was the most active, but most of the other fractions were also quite active. Structural studies showed that the most active fraction was a pectin of the rhamnogalacturonan type 1 with side chains of the arabinogalactan type II. Removal of terminal arabinofuranosides lead to a drop of the activity indicating that this structural unit may be involved in the bioactive site of the molecule. PMID- 12648828 TI - Sustainable harvest of medicinal plants at Bulashbar Nullah, Astore (Northern Pakistan). AB - Rapid decline of plant resources due to their conventional use needs ex-situ and in-situ conservation, training of the community regarding collection of medicinal plants and their marketing. In this regard, the Bulashbar valley, Astore, District Diamer was identified as a case study. The main objectives of this activity were to enlist economic, medicinal and aromatic plants including their occurrence, general distribution and abundance in the project areas; to determine traditional use and pharmaceutical values of each medicinal plant species found in the project area. Ethnobotanical studies of the area revealed that 33 plants were being used by the local communities for medicinal purposes. Two species, Bunium persicum and Ephedra gerardiana, are recommended for in vitro cultivation to obtain quick benefits. While Hippophae rhamnoides can be sustainably used for socio-economic uplift of the local communities. PMID- 12648829 TI - Antihypertensive, antiatherosclerotic and antioxidant activity of triterpenoids isolated from Olea europaea, subspecies africana leaves. AB - For the first time a biossay-directed study of triterpenoids isolated from the leaves of Olea europaea from Greece, from wild African olive and from a cultivar of O. europaea grown in Cape Town was reported. The experiment was undertaken since our preliminary analyses showed that the African wild olive leave is rich in triterpenoids and contain only traces of the glycoside oleuropein, which is typical for the European olive leaves. The isolate of the African wild olive leaves (AO) used in the experiments was found to contain 0.27% 1:1 mixture of oleanolic acid and ursolic acid, named oleuafricein. The isolate of Greek olive leaves (GO) was found to contain 0.71% oleanolic acid, and the Cape Town cultivar (CT) contained 2.47% oleanolic acid. No ursolic acid was found in either GO or CT. The antihypertensive, diuretic, antiatherosclerotic, antioxidant and hypoglycemic effects of authentic oleanolic and ursolic acid and the three isolates (GO, AO and CT) were studied on Dahl salt-sensitive (DSS), insulin resistant rat genetic model of hypertension. All three isolates, in a dose 60 mg/kg b.w. for 6 weeks treatment, prevented the development of severe hypertension and atherosclerosis and improved the insulin resistance of the experimental animals. GO, OA and CT isolates could provide an effective and cheap treatment of this particular, most common type of salt-sensitive hypertension in the African population. PMID- 12648830 TI - Evaluation of antidiarrhoeal activity Rumex maritimus root. AB - The partitioned n-hexane, ethylacetate and residual methanol extracts of the root of Rumex maritimus were evaluated for antidiarrhoeal activity in mice using castor oil and serotonin induced diarrhoea and also charcoal motility test at 50, 100 and 200 mg/kg body weight dose. Among these, the methanol extract most significantly prolonged the time for induction of diarrhoea, reduced the frequency of diarrhoeal episodes and also decreased the propulsion of charcoal meal through the gastrointestinal tract. PMID- 12648832 TI - Proceedings of the FEMS-Symposium on The Versatility of Listeria Species. Izmir, Turkey, October 10-11, 2002. PMID- 12648833 TI - Listeriosis: clinical presentation. AB - Listeria monocytogenes is an uncommon cause of illness in the general population. However, this bacterium is an important cause of severe infections in neonates, pregnant women, the elderly, transplant recipients and other patients with impaired cell-mediated immunity. Various clinical syndromes due to L. monocytogenes have been described such as sepsis, central nervous system infections, endocarditis, gastroenteritis and localized infections. A review of the clinical presentation of listeriosis is given in this paper. PMID- 12648834 TI - Murine model of pregnancy-associated Listeria monocytogenes infection. AB - Listeria monocytogenes has been recognized as a significant pathogen, occurring worldwide, capable of causing animal and human infections. In its most severe form, listeriosis is an invasive disease that affects immunocompromised patients. Additionally, pregnant women represent a high-risk group for L. monocytogenes infection. Abortion, stillbirth or severe neonatal infection can be the serious outcome of such an infection. In an experimental murine model of pregnancy associated listeriosis we studied the impact of L. monocytogenes on the maternal immune response and pregnancy outcome. In comparison to virgin animals, pregnant mice mounted lower levels of protective cytokines and were unable to eliminate the pathogen. The impaired maternal immune response that has been found both on the systemic and local level, facilitated bacterial multiplication in the liver, placenta and ultimately in the fetal tissues. This resulted in severe necrotizing hemorrhagic hepatitis and Listeria-induced placental necrosis, increasing the incidence of postimplantation loss and poor pregnancy outcome. PMID- 12648835 TI - Listeria: growth, phenotypic differentiation and molecular microbiology. AB - The identification of Listeria species is based on a limited number of biochemical markers, among which absence or presence of hemolysis and arylamidase are used to differentiate between L. monocytogenes and L. innocua. The CAMP (Christie, Atkins, Munch-Petersen) test must be interpreted with caution. Chromogenic media are based on both the specific chromogenic detection of phosphatidylinositol phospholipase C and the xylose fermentation and give specific and direct identification of L. monocytogenes and L. ivanovii. Isolates of L. monocytogenes with atypical properties require tools of molecular biology for final identification. Serotyping, although not allowing speciation, serves a useful purpose for confirming the genus diagnosis Listeria. Polymerase chain reaction is particularly useful when prior administration of antimicrobial agents compromises culture. For clinical specimens the importance of trying to isolate the pathogen as a prerequisite for an epidemiological work-up and finally for prevention of further cases cannot be overstressed. PMID- 12648836 TI - Listeria monocytogenes: diagnostic problems. AB - The first isolation methods for the detection of Listeria spp. were generally based on the direct culture of samples on simple agar media, but isolation of the pathogenic Listeria monocytogenes was difficult. In time, new techniques were developed, based on a variety of selective and elective agents in isolation and enrichment media, which gained better and quicker results. Current reference methods allow the recovery of L. monocytogenes from a variety of foods with relative ease. However, more comparative studies are needed to select one horizontal method. It is suggested that the procedure of the International Organization for Standardization is a good base for such comparisons. PMID- 12648837 TI - History and epidemiology of listeriosis. AB - Listeriae are used as a tool by different specialities in biomedical research. There are now at least four major fields of interest in Listeria: (1). the role in medical microbiology: Listeria monocytogenes causes severe diseases of men and animals and is difficult to treat; (2). the role in food microbiology: Listeria is a food-borne pathogen and is found in various food items; (3). the role in cell biology: L. monocytogenes is a facultative intracellular parasite having an intense cross-talk and interactions with the host cell; (4). the role in immunology: basic knowledge on cell-mediated immunity has been acquired through the model of listeriosis. This paper presents information on the past and the actual situation in research on Listeria and listeriosis. PMID- 12648838 TI - Listeriosis: therapeutic options. AB - Since overt listeriosis occurs mainly in immunocompromised persons it is quite consistent to try to restore the hampered defence system by supportive measurements. For direct antimicrobial treatment a series of different antibiotics is available, since Listeria strains isolated from patients are in general susceptible to a wide range of antibiotics, except fosfomycin, quinolones and cephalosporins of the third generation, although a few exceptional strains exist. Unfortunately, most antibiotics are not bactericidal for Listeria. Drug combinations may exert a synergistic effect. Furthermore, the efficacy of therapy is limited by the fact of intracellular habitat of pathogenic Listeria. Few agents, such as macrolides and quinolones, are accumulated within host cells and may attack the intracellular Listeriae. The clinical experience shows that the combination of amoxicillin and gentamicin is the best option. PMID- 12648839 TI - Comparison of the genome sequences of Listeria monocytogenes and Listeria innocua: clues for evolution and pathogenicity. AB - Listeria monocytogenes, an invasive opportunistic, food-borne pathogen, remains one of the leading causes of mortality from food-borne infections. The recently determined complete genome sequences of L. monocytogenes strain EGDe and of that of the closely related non-pathogenic species Listeria innocua strain CLIP11262 enhance our knowledge of the genetic basis of the virulence of L. monocytogenes and advance our understanding of the evolution of these Listeria species. Both genomes encode a high number of surface, transport and regulatory proteins. Comparison of the genome organisation revealed a perfect synteny between the two Listeria genomes. Comparison with other closely related bacteria also showed a high conservation in genome organisation among the Listeria, Staphylococcus and Bacillus group of low G+C content bacteria. Distinct G+C content of a number of strain-specific genes suggests intensive lateral gene transfer. The identification of a 55-kb locus encoding proteins with high homology to Salmonella enterica serovar Typhimurium vitamin B(12) synthesis proteins as well as those necessary for degradation of ethanolamine and propanediol further indicates acquisition of a complete metabolic pathway by horizontal gene transfer and a probable role of this locus in anaerobic growth in the host. PMID- 12648840 TI - Nucleic acid-based, cultivation-independent detection of Listeria spp and genotypes of L monocytogenes. AB - Based on comparative analysis of 16S rRNA gene sequences, two oligonucleotide probes for in situ detection of all members of the genus Listeria were designed. These probes allowed fast and reliable in situ detection of Listeria spp. even in complex samples like raw milk. Almost full-length iap (invasion-associated protein) gene sequences were determined for 69 Listeria monocytogenes strains of all 13 known serotypes. A comparison of these sequences revealed that the L. monocytogenes strains can be grouped into three distinct genotypes. These clusters correlate well with distinct serotypes. Thus, strains of serotypes b and d belong to genotype I, a and c to genotype II, and 4a and 4c, which are rarely isolated from humans, group together within genotype III. These results could be corroborated by further comparative sequence analysis of genes encoding two phospholipases - plcA and plcB. Based on the iap gene sequences, a highly specific and reproducible competitive PCR detection method was developed. Primer pairs targeting genotype-specific regions of the iap gene were designed. The amplification of non-specific PCR products from DNA of non-target strains was prevented by adding competitive primers. By applying this method, the rapid and reliable distinction of the three L. monocytogenes genotypes was possible. PMID- 12648841 TI - Characterization of Listeria monocytogenes recovered from 41 cases of sporadic listeriosis in Austria by serotyping and pulsed-field gel electrophoresis. AB - 41 clinical Listeria monocytogenes strains recovered from seven feto-maternal and 34 non-pregnancy associated cases of human listeriosis documented between 1997 and 2000 underwent serotyping and typing by pulsed-field gel electrophoresis (PFGE) applying the enzymes AscI, ApaI and SmaI. The pulsotypes of the clinical strains were compared to the pulsotypes of three L. monocytogenes strains isolated from healthy fecal carriers and nine reference strains isolated from seven outbreaks in Europe and the USA. The 41 clinical strains of Austrian provenance showed 37 pulsotypes. Five sets of two Austrian strains each were indistinguishable by PFGE typing. Epidemiological links were absent between these indistinguishable isolates. One unique pulsotype (AB) was found in three fecal isolates. Five pulsotypes (A, Q, R, AC and AD) were distinguished among the strains associated with outbreaks. Clusters consisting of two, five and six Austrian strains each were indistinguishable from the outbreak-associated pulsotypes A, Q and R, respectively, after PFGE analysis with AscI. Three strains of AscI pulsotype Q and five strains of AscI pulsotype R could be further differentiated by restriction with ApaI and SmaI. One strain each from sporadic cases shared a combined pulsotype with the outbreak strains of pulsotypes A and R, respectively. These PFGE data suggest that a similar genetic background can be found in strains which have been contributing to outbreaks world-wide and in isolates associated with sporadic listeriosis in Austria. PMID- 12648842 TI - CD8 T cell immunome analysis of Listeria monocytogenes. AB - The identification of T cell epitopes is crucial for the understanding of the host response during infections with pathogenic microorganisms. Generally, the identification of relevant T cell responses is based on the analysis of T cell lines propagated in vitro. We used an ex vivo approach for the analysis of the CD8 T cell response against Listeria monocytogenes that is based upon the fractionation of naturally processed antigenic peptides and subsequent analysis with T cells in an enzyme-linked immunospot (ELISPOT) assay. Our data indicate that the direct ex vivo ELISPOT analysis of peptides extracted from infected tissues represents a versatile and potent test system for the analysis of the CD8 T cell immunome of microorganisms that furthermore requires neither the knowledge of the microbial genome nor of the specificity of responding T cells. PMID- 12648843 TI - Tailoring host immune responses to Listeria by manipulation of virulence genes -- the interface between innate and acquired immunity. AB - Although attenuated strains of microbial pathogens have triggered vaccine development from its origin, the role of virulence factors in determining host immunity has remained largely unexplored. Using the murine listeriosis model, we investigated whether the induction and expansion of protective and inflammatory T cell responses may be modified by selective manipulation of virulence genes. We intentionally deleted specific genes of Listeria monocytogenes, including those encoding the positive regulatory factor (prfA), hemolysin (hly), the actin nucleator (actA), and phospholipase B (plcB). The resulting strains showed decisive differences in their immunogenic properties. In particular, we identified a double-deletion mutant that retained Listeria's profound ability to induce protective CD8(+) T cells, but that is strongly attenuated and exhibits a significantly reduced ability to induce CD4(+) T cell-mediated inflammation. We conclude that this mutant, L. monocytogenes DeltaactADeltaplcB, is at present the most promising mutant for a bacterial vaccine vector and is able to safely induce potent CD8(+) T cell-mediated immunity. PMID- 12648845 TI - Quantitative risk assessment of Listeria monocytogenes in ready-to-eat foods: the FAO/WHO approach. AB - Quantitative microbiological risk assessment is a very new and unique scientific approach able to link, for the first time, data from food (in the farm-to-fork continuum) and the various data on human disease to provide a clear estimation of the impact of contaminated food on human public health. The Food and Agriculture Organization of the United Nations (FAO) and the World Health Organization (WHO) have recently launched risk assessment studies of a number of pathogen-food commodity combinations (Salmonella in eggs and in broiler chickens, Listeria monocytogenes in ready-to-eat foods, Campylobacter in broiler chickens, Vibrio in seafood) to be used to lower the risk associated with these food-borne diseases and ensure fair practices in the international trade of food. The FAO/WHO Listeria risk assessment was undertaken in part to determine how previously developed risk assessments done at the national level could be adapted or expanded to address concerns related to L. monocytogenes in ready-to-eat foods at an international level. In addition, after initiation of the risk assessment, the risk assessors were asked by the Codex Committee on Food to consider three specific questions related to ready-to-eat foods in general, which are: (1). estimate the risk for consumers in different susceptible populations groups (elderly, infants, pregnant women and immunocompromised patients) relative to the general population; (2). estimate the risk for L. monocytogenes in foods that support growth and foods that do not support growth under specific storage and shelf-life conditions; (3). estimate the risk from L. monocytogenes in food when the number of organisms ranges from absence in 25 g to 1000 colonies forming units per gram or milliliter, or does not exceed specified levels at the point of consumption. To achieve these goals, new dose-response relationships and exposure assessments for ready-to-eat foods were developed. Preliminary data indicate that eliminating the higher dose levels at the time of consumption has a large impact on the number of predicted cases. PMID- 12648846 TI - Efficiency of high pressure treatment for destruction of Listeria monocytogenes in fruit juices. AB - The objective of this study was to compare high pressure resistance of Listeria monocytogenes strains at 25 degrees C and 50 degrees C at 350 MPa and to use high pressure (250 MPa and 350 MPa) at 30 degrees C and 40 degrees C for the inactivation of the relatively most pressure resistant strain inoculated in pasteurized apple, apricot, cherry and orange juices. L. monocytogenes CA was found to be the relatively most pressure resistant strain and increasing pressurization from 250 MPa to 350 MPa at 30 degrees C had an additional three to four log cycle reduction in viability, still leaving viable cells after 5 min. When 350 MPa at 40 degrees C for 5 min was applied more than eight log cycle reduction in cell population of all fruit juices was achieved. This study demonstrated that low temperature (40 degrees C) high pressure (350 MPa) treatment has the potential to inactivate relatively pressure resistant L. monocytogenes strains inoculated in different fruit juices within 5 min. PMID- 12648844 TI - Production of IL-12 and IL-18 in human dendritic cells upon infection by Listeria monocytogenes. AB - Dendritic cells (DCs) are major antigen-presenting cells of the immune system, which need to be activated in order to initiate an immune response. Here, we describe the immunostimulatory effects on human monocyte-derived DCs observed upon infection with Listeria monocytogenes or after treatment with listerial lipoteichoic acid (LTA) and lipopolysaccharide (LPS), respectively. All stimuli caused upregulation of costimulatory molecules, induced T-cell proliferative responses and secretion of cytokines in vitro. Infection of DCs with L. monocytogenes induced release of interleukin (IL)-12 and IL-18. In contrast treatment with purified listerial LTA yielded high levels of IL-18 release, but only minimal IL-12 production. Treatment of DCs with LPS conversely induced significant amounts of IL-12 production, but no IL-18. The release of both stimulating cytokines IL-12 and IL-18 upon infection with entire bacteria suggests that attenuated strains of L. monocytogenes may be a valuable tool for subunit vaccine delivery. PMID- 12648847 TI - Occurrence of Listeria sp and L monocytogenes in sewage sludge used for land application: effect of dewatering, liming and storage in tank on survival of Listeria species. AB - The application of sewage sludge to agricultural land is widely used in France. To determine the impact of sludge treatments, concentrations of Listeria sp., Listeria monocytogenes and faecal indicators were monitored in five types of sludge from three sewage treatment plants in Angers (France) and its suburbs over a 1-year period. On the whole, bacteria were reduced in numbers through sludge treatments. Apart from liming, which leads to reduced levels of bacteria below detection limits, other sludge treatments did not eliminate Listeria sp. and faecal indicators. Listeria sp. and L. monocytogenes were found respectively in 87% and 73% of dewatered sludges and in 96% and 80% of sludges stored in tanks. Concentrations of L. monocytogenes, ranging from 0.15 to 20 MPN g(-1) dry matter in dewatered sludge and from 1 to 240 MPN g(-1) dry matter in sludge stored in tanks, did not show seasonal variations. Spreading of sanitised sludge onto agricultural land results in the addition of 10(6)-10(8) L. monocytogenes per hectare per year, which may contribute to the increase in the dissemination of this pathogenic species in the environment. PMID- 12648848 TI - A mathematical model accounting for the organization in multiplets of the genetic code. AB - A model using suitable mathematical operators in the crystal basis model of the genetic code is presented. This model retains a requirement for stability of the genetic code against misreading or translation errors. The main features (including number of encoded amino-acids, nucleotide content, and synonymous codons multiplet dimension) are described for mitochondrial and eukaryotic genetic codes. PMID- 12648849 TI - Mathematical formulation of Leibnizian world: a theory of individual-whole or interior-exterior reflective systems. AB - A world model, suggested by Leibniz's monadology, is formulated as a mathematical axiomatic system. The purpose of this world model is to provide a general scheme for describing a system of individuals having consciousness or internal worlds that communicate with each other and make a unified whole world, and moreover, the latter is reflected into the respective internal worlds and appears as an external world. Examples of such monadological structure of interior-exterior (or individual-whole) reflection can be observed in bio- or socio-systems and recently in computer networks. Moreover, a most elemental version of this structure can be found in the basic level of quantum physics. The model not only gives a prototype of monadological systems but also has an evolutionary ability to produce a hierarchy of monadological systems, which are interpreted as corresponding to various levels of consciousness. PMID- 12648850 TI - Quantum monadology: a consistent world model for consciousness and physics. AB - The NL world model presented in the previous paper is embodied by use of relativistic quantum mechanics, which reveals the significance of the reduction of quantum states and the relativity principle, and locates consciousness and the concept of flowing time consistently in physics. This model provides a consistent framework to solve apparent incompatibilities between consciousness (as our interior experience) and matter (as described by quantum mechanics and relativity theory). Does matter have an inside? What is the flowing time now? Does physics allow the indeterminism by volition? The problem of quantum measurement is also resolved in this model. PMID- 12648851 TI - Quantitative measures of organization for multiagent systems. AB - A set of "information theoretic" measures has been developed to quantify the degree of constraint inherent in the organization of a multiagent system. Separate measures can be provided to quantify spatial organization, trophic organization and, more generally, the overall structure of interactions. The additive character of these quantities allows them to be distributed in various fashions among species and places in a way that allows one to assign an "Importance Index" to those taxa and places. In addition, a measure to gauge the degree of adaptation of a species to a particular environment is proffered. The proposed measures allow one to formulate the following hypotheses in quantitative fashion: (1). that any disturbance of an ecosystem at a location associated with a high spatial Importance Index will exert a greater impact on the population dynamics than will a similar disturbance aimed at a place where the values of these indexes are lower; (2). that any disturbance in an ecosystem affecting a particular species with high individual Importance Indexes will cause a greater impact on the overall population dynamics than will a disturbance aimed at a species with a lower values of these indexes; (3). that the ascendancy of evolving system has a propensity to increase. The precise quantitative formulation of these hypothesis would permit them to be tested via multiagent simulation. Estimating the probablities pertaining to these hypotheses presents a number of problems that merit discussion. PMID- 12648853 TI - How to decide whether small samples comply with an equidistribution. AB - The decision whether a measured distribution complies with an equidistribution is a central element of many biostatistical methods. High throughput differential expression measurements, for instance, necessitate to judge possible over representation of genes. The reliability of this judgement, however, is strongly affected when rarely expressed genes are pooled. We propose a method that can be applied to frequency ranked distributions and that yields a simple but efficient criterion to assess the hypothesis of equiprobable expression levels. By applying our technique to surrogate data we exemplify how the decision criterion can differentiate between a true equidistribution and a triangular distribution. The distinction succeeds even for small sample sizes where standard tests of significance (e.g. chi(2)) fail. Our method will have a major impact on several problems of computational biology where rare events baffle a reliable assessment of frequency distributions. The program package is available upon request from the authors. PMID- 12648852 TI - A context-free data compression approach to measuring information transmission by action potentials. AB - Action potentials allow nervous systems to transmit information rapidly and efficiently over considerable distances, but what is the information they carry and how much can be carried by one neuron? Often, qualitative and vague descriptions are used, such as the firing rate representing intensity. Recent attempts to quantify information transmission by action potentials have concentrated on treating neurons as communication channels, whose information capacity can be estimated from their signal-to-noise ratios. However, this only indicates how much information could theoretically be carried, not the actual amount at any given time, and the ratio itself depends on assumptions about information coding. Here we introduce a different approach based on the concept of data compression, which has become familiar with the widespread use of digital computers and networks. Compression takes advantage of redundancy in a sequence of numbers to reduce its size, but allows it to be reconstructed later without error. We show that data compression by a context-free grammar can quantitatively estimate the real information content of action potential signals without any prior assumptions about coding, or knowledge of neural inputs. Measurements of information coding by mechanosensory neurons are used as examples, but a major advantage of this approach is its generality. It can estimate information transmission by any neuron whose output can be measured, regardless of neuronal type, connectivity or function. PMID- 12648855 TI - Postmenopausal hormone replacement therapy in the light of the women's health initiative trial. PMID- 12648856 TI - Homocysteine, folic acid and B-group vitamins in obstetrics and gynaecology. AB - The amino acid homocysteine is of considerable medical importance because it is involved in the etiopathogeny of cardiovascular diseases. Hyperhomocysteinemia is a factor in the vascular damage that predisposes thrombogenesis and arteriosclerosis. It has also been related to a range of obstetric and gynecologic complications. Numerous studies have demonstrated the association between increased levels of homocysteine and neural tube defects (NTD) and other congenital defects, spontaneous miscarriages, intrauterine growth retardation (IGR), preeclampsia and intrauterine fetal death. Folic acid and other B-group vitamins (B6 and B12) take part in the metabolism of homocysteine and the preventive administration of these vitamins may reduce some of the complications. PMID- 12648857 TI - Trends in maternal mortality: a 13-year hospital-based study in rural Ghana. AB - OBJECTIVE: To measure the impact of the Safe Motherhood Initiative (SMI) on hospital-based maternal mortality since its start in 1987. STUDY DESIGN: Retrospective analysis of all 229 maternal deaths in a district hospital in rural Ghana, between 1 January 1987 and 1 January 2000. Main outcome measures were maternal mortality ratio and relative contribution of causes of maternal deaths to overall maternal mortality. Chi-square test was used to assess differences in proportions, and relative risks with confidence intervals were calculated. RESULTS: The overall maternal mortality ratio of 1077/100,000 live births did not change significantly during the study period. However, the relative contributions of sepsis, hemorrhage, obstructed labor, anemia/sickle cell disease and (pre-) eclampsia diminished, while abortion complications increased significantly. CONCLUSIONS: The Safe Motherhood Initiative in the study area has contributed to the reduction of maternal mortality due to causes against which interventions were directed. Abortion complications as cause of maternal mortality need to be included in interventions and research. PMID- 12648858 TI - Six hundred and ten breech versus 12,405 cephalic deliveries at term: is there any difference in the neonatal outcome? AB - OBJECTIVES: To compare neonatal morbidity of breech and cephalic deliveries at term. STUDY DESIGN: Cohort study of 610 consecutive singleton breech presentations and 12,405 consecutive singleton cephalic presentations in term between 1992-1998. Five hundred and fourteen breech and 11,989 cephalic presentations were candidates for vaginal delivery, of which 407 (79%) breeches and 11,265 (94%) cephalic delivered vaginally. RESULTS: Neonatal intensive care admissions were significantly greater for breech than cephalic vaginal deliveries (2.7% versus 0.25%, P = 0.000), but newborn intensive care admission and mortality were equally distributed between the two groups. CONCLUSIONS: A low caesarean rate is possible (21% beech and 6% cephalic). Neonatal morbidity was equal in the two populations. Admission to neonatal intensive care was significantly more frequent for caesarean section than for vaginal delivery in the cephalic group and equal in the breech group. This study justifies our obstetrical policy and the realisation of a trial in several centres similar in terms of perinatal management. PMID- 12648859 TI - Pregnancies complicated by Twin-Twin transfusion syndrome: outcome and long-term neurological follow-up. AB - OBJECTIVE: To relate prenatal data of monochorionic pregnancies complicated by Twin-Twin transfusion syndrome (TTTS) with survival rates and neurological morbidity. STUDY DESIGN: Thirty-two cases of TTTS underwent biweekly ultrasound examinations. Amnioreduction was the standard of care. Mortality and neurological morbidity were evaluated; the children had neurodevelopmental follow-up with a mean follow-up time of 24 months. RESULTS: Ten out of 32 pregnancies terminated before 24 weeks of gestation, six after induction and four after spontaneous abortion. Among the 22 pregnancies followed up after 24 weeks, the overall survival rate at 28 days was 70% (31/44) with 10 intrauterine deaths including three cases of single selective terminations, and with three neonatal deaths. Eighteen out of 31 babies (58%) had a normal neurological development, eight cases had major and five cases minor neurological disabilities. Neonatal survival was not related to gestational age at diagnosis, number of amnioreductions or weeks at delivery. Absence of end diastolic flow in the umbilical artery, either in the donor or the recipient twin, and birth weight <1000 g were both associated with a higher perinatal mortality. Number of amnioreductions (>2) and birth weight <1000 g were both associated with abnormal neurological follow-up. CONCLUSION: In TTTS, absent end diastolic flow in the umbilical artery and birth weight can predict perinatal survival; neurological development is correlated with number of amnioreductions and birth weight. PMID- 12648860 TI - What to do after a failed attempt of vacuum delivery? AB - OBJECTIVES: To determine whether there is a difference in maternal and neonatal outcomes if a sequential operative vaginal or cesarean delivery follows failed vacuum delivery. STUDY DESIGN: A cross sectional study. We have analyzed maternal and neonatal outcomes of 215 vacuum extractions (group 1), 106 forceps assisted deliveries (group 2), 28 deliveries in which failed vacuum extraction were followed by forceps delivery (group 3) and 22 deliveries in which failed vacuum extraction were followed by cesarean delivery (group 4). RESULTS: Compared to other groups, patients in group 4 had significantly more post partum anemia, meconium stained amniotic fluid and hospital stay (both maternal and neonatal) as well as lower pH. Apgar scores were similar in groups 3 and 4. Incidence of respiratory distress syndrome, cephalhematoma and jaundice were similar in neonates of all groups. CONCLUSIONS: If an attempted vacuum delivery has failed, the risk of adverse neonatal outcome is increased with either subsequent forceps or cesarean delivery. It should remain in the judgment of the attending obstetrician to choose the method most suitable under the given circumstances. PMID- 12648861 TI - Reference values and predictors of white blood cell subset counts: a cross sectional study among HIV seronegative pregnant women in Zimbabwe. AB - OBJECTIVE: To identify predictors and develop reference values of white blood cell subset counts for pregnant black women in Zimbabwe. STUDY DESIGN: In this cross-sectional study, multiple linear regression (MLR) analysis was employed to assess the relationship of WBC subset counts with age, gestational age, gravidity, season, serum retinol, beta-carotene, ferritin, folate and alpha-1 antichymotrypsin among 998 women 22-35 weeks pregnant attending antenatal care (ANC) in Harare, Zimbabwe. RESULTS: Mean age was 24.0 (95% CI; 23.6-24.4), range 14-45 years. The mean gestational age was 29.2 (95% CI; 29.0-29.4), range 22-35 weeks. Median gravidity was 2, range 1-9. Predictors of neutrophil counts were gestational age, season and serum ferritin, the latter in interaction with gravidity (interaction, p = 0.016). Mean lymphocyte count was 0.13 x 10(9)cells/l higher in gravida >4 than gravida 1-3, and 0.35 x 10(9)cells/l higher in the late rainy than other seasons. Predictors of monocyte counts were gestational age, serum folate and season, while eosinophil counts declined with advancing gestation. Reference values adjusted or unadjusted for identified predictors were different from those of pregnant and non-pregnant white women reported in the literature. CONCLUSIONS: Gravidity, season and micronutrient status influence WBC counts during pregnancy and therefore are of physiological and clinical importance. WBC reference values in the literature were not applicable obviating the need for local reference values. PMID- 12648862 TI - Tocolytic effects of formoterol are associated with local change in the progesterone/estradiol ratio. AB - OBJECTIVE: The aim of the study was to determine whether a beta(2)-adrenoceptor agonist, formoterol, inhibits premature delivery in connection with change in estradiol and progesterone concentrations in the amniotic fluid in ovariectomized rats. STUDY DESIGN: Pregnant rats at the 15th day of gestation were bilaterally ovariectomized and given injection of 17beta-estradiol immediately after the operation and every 24 h. An osmotic pump filled with a solution of formoterol or saline was also implanted subcutaneously into the back of each. The animals were killed by decapitation under light ether anesthesia 18, 36, 54 or 72 h after ovariectomy, and the numbers of undelivered fetuses and newborn were counted. Amniotic fluid was collected 16, 36, and 54 h after ovariectomy. RESULTS: Formoterol (0.15 mg/(kg h)) reversed the decline in premature delivered fetuses due to 17beta-estradiol 54 and 72 h after ovariectomy. Although no influence was evident regarding the progesterone and estradiol concentrations in amniotic fluid in ovariectomized rats supplemented with 17beta-estradiol, formoterol significantly inhibited the increment in the estradiol/progesterone ratio as well as the elevation in prostaglandin F2alpha concentration. CONCLUSION: These findings indicate that tocolytic effects of formoterol may be associated with suppression of uterine activity due to modulation of hormone secretion. PMID- 12648863 TI - Local and systemic tolerability of magnesium sulphate for tocolysis. AB - An open-label, randomised, parallel-group, study was conducted in three study centres in women with premature labor and indication for a single agent intravenous tocolysis therapy with magnesium sulphate. The aim of this study was to examine the local and general tolerability and side-effects of magnesium sulphate for tocolysis. Furthermore, we tested the tolerability of a ready-for use magnesium solution. No measurements of efficacy were performed during this study. Initially, patients received a loading dose of 4.0 g magnesium sulphate administered over 30 min. Thereafter, a continuous intravenous infusion of 1-2 g magnesium sulphate per hour up to 21 days was given. Venous score (Maddox), vital signs, adverse events as well as general tolerability (assessed by investigator and patients) and blood parameters were assessed. We showed good local and systemic tolerability of high dose magnesium sulphate for tocolysis. Only seven patients (15%) were withdrawn from the study prematurely due to minor adverse events. Potential serious complications of MgSO(4) such as respiratory arrest or clinically relevant respiratory depression were not observed. The most frequently reported local adverse events were injection site pain, itching, erythema, swelling, induration and palpable venous cord. The most common systemic adverse events considered to be possibly related to the study drugs involved the nervous system (dizziness) followed by the digestive system (nausea, constipation). Systolic and diastolic blood pressure changed only slightly during the treatment. Respiratory rate and body temperature remained stable also. Toxic magnesium levels (>2.5 mmol/l) were not observed. The assessment of the clinical investigators with regard to tolerability was very good or good in 72.5% of the patients. The introduction of the ready-to-use solution has the advantage of eliminating the need to mix the solution prior to administration. This means a lower risk of overdose and contamination. PMID- 12648864 TI - Pregnancy rate is not improved by delaying embryo transfer from days 2 to 3. AB - OBJECTIVE: To compare the outcome of assisted reproduction in day 2 versus day three embryo transfer. DESIGN: Prospective study. PARTICIPANTS: A total of 927 consecutive embryo transfers for IVF and ICSI cycles including 626 embryo transfers on day 2 and 301 on day 3. INTERVENTION: IVF and ICSI. OUTCOME MEASURE: Clinical pregnancy rate. RESULTS: There is no significant difference in the pregnancy rate between ET on day 2 (50.9%) and ET on day 3 (50.5%). CONCLUSION: Embryo transfer could be done on days 2 or 3 according to the convenience of the patient and the medical team. CONDENSATION: Embryo transfer could be done on days 2 or 3 according to the convenience of the medical team with similar results. PMID- 12648865 TI - The effect of sodium monofluorophosphate therapy on lipid and lipoprotein metabolism in postmenopausal women. AB - OBJECTIVE: To investigate the effect of a low dose of fluoride, combined with hormone replacement therapy (HRT) or alone, on serum lipids and lipoproteins in postmenopause. STUDY DESIGN: One hundred and thirty-five healthy postmenopausal women were enrolled in this prospective study and randomly assigned to: (1) oral monofluorophosphate (MFP, 25 mg per day); or (2) HRT (a combination of oral conjugated estrogen 0.625 mg in conjunction with medroxyprogesterone acetate 5 mg daily); or (3) HRT + MFP; or (4) placebo, for 18 months. Serum lipid and lipoprotein levels were measured at enrollment and at the end of the study period. RESULTS: Total cholesterol was not significantly different among groups, with a decrease in the HRT and the HRT + MFP groups (respectively, -7.8 and 7.9%), and a small increase in the MFP group (0.4%). LDL-C did not differ significantly in the groups, with a decrease in the HRT and HRT + MFP groups (respectively, -10.3 and -10.4%), and a slight increase in the MFP group (0.8%). TG decreased with -12 and 11.8% in the HRT and HRT + MFP groups (P = 0.052 and 0.055, respectively), but a slight increase was seen in the MFP group (1.4%). For HDL-C, both HRT and HRT+MFP groups showed a small increase (respectively, 3.5 and 3.6%), whereas MFP group showed a slight decrease (-0.9%, NS). CONCLUSION: We conclude that long-term use of a low dose of fluoride had no significant adverse changes in the lipid and lipoprotein profile in postmenopause. It neither attenuated nor potentiated the effect of HRT on lipids and lipoproteins. PMID- 12648866 TI - Voiding symptoms in chronic pelvic pain (CPP). AB - OBJECTIVE: To establish the prevalence of voiding symptoms in CPP patients. STUDY DESIGN: Sixty women with chronic pelvic pain (CPP), aged 23-79 (mean: 48) years completed a questionnaire mainly addressing lower abdominal pain and voiding symptoms. Symptoms occurring often, almost always, or always for more than 6 months were considered present and serious. A group of 31 consecutive cases of non-CPP women aged 18-77 (mean: 49) years were selected to serve as controls. RESULTS: The following symptoms were found in the CPP group: incontinence: 43%; inadequate voluntary control of the urethral sphincter: 50%; inability to postpone: 37%; urge: 37%; nocturia > or =2X: 18%; dysuria: 12%; cystitis: 37%; urge-induced pain: 20%; pain-induced urge: 18%; strain to initiate voiding: 6%; strain to continue voiding: 17%; incomplete voiding: 37%. Two or more voiding symptoms were present in 63% of the CPP group. All urinary symptoms were more often present in the CPP group than in the controls. CONCLUSION: A substantial subgroup of CPP patients has voiding symptoms. PMID- 12648867 TI - Severity of symptoms in primary dysmenorrhea--a Doppler study. AB - OBJECTIVE: Doppler findings in women with severe symptoms of primary dysmenorrhea include high impedance to blood flow in uterine arteries with a preserved cyclic pattern throughout the whole cycle. Doppler findings in women who present with mild symptoms of primary dysmenorrhea are not yet documented. The aim of this study was to investigate possible differences in Doppler findings among women with mild and severe primary dysmenorrhea. STUDY DESIGN: One hundred and fifty four women were examined with color Doppler ultrasound: 50 in the control group, 60 in the mild and 44 in the severe primary dysmenorrhea subgroup. We calculated resistance index in uterine arteries in these women on the first day of the cycle, in the follicular (days 9-12) and the luteal (days 20-23) phase of the cycle and used analysis of variance for comparing results. RESULTS: The rate of visualization was 100% for uterine and arcuate arteries, 44-76% for the radial and 32-62% for spiral arteries, respectively. A significant difference in Doppler index values among the mild and severe dysmenorrheic group was observed in the luteal phase for the arcuate artery and in all the three measurement periods for the radial and spiral arteries. CONCLUSION: There is a difference in Doppler findings between women with mild and severe symptoms of primary dysmenorrhea. PMID- 12648868 TI - Laparoscopic surgery in women with endometrial cancer: the learning curve. AB - OBJECTIVE: The aim of this study was to assess the effect of increasing surgeons's experience in the laparoscopic surgery of women with endometrial cancer (EC) on the surgical outcome of these patients. STUDY DESIGN: Data were obtained from a prospectively collected database of 108 patients two oncolaparoscopic centers in Czech Republic who underwent laparoscopically assisted surgical staging (LASS) from April 1996 to March 2001. Patients were arranged in chronological order and divided into three groups, based on the date of their surgery. The three groups were compared in patient characteristics and surgical outcome using one-way analysis of variance (ANOVA) and Wilcoxon rank sum test. SETTING: Department of Obstetrics and Gynecology, Endoscopic Training Center, Baby Friendly Hospital Kladno, Czech Republic. RESULTS: The three groups were similar in patient characteristics. Operative times for laparoscopic staging with pelvic lymphadenectomy (LN) decreased significantly from mean of 156.3 min for group 1 to 142.8 min for group 3 (P < 0.05). In cases LASS with pelvic lymphadenectomy was significant increase in the number lymph nodes harvested (12.4 for group 1, 13.9 for group 2, and 15.4 for group 3, P < 0.05). In cases LASS without lymphadenectomy was not significant difference in operating time, estimated blood loss, rate of conversion to laparotomy, operative complications, and length of hospital stay among the compared groups. The number of patients who underwent para-aortic lymphadanectomy was too small (n = 22), and their distribution was asymmetrical for comparison. CONCLUSION: A learning curve is demonstrated in the LASS of women with endometrial cancer. With increasing surgeon's team experience, there is significant decrease in operative time for staging with pelvic lymph node dissection and increase in the number of pelvic lymph nodes removed. The para-aortic lymphadenectomy (PALN) was found to be more challenging than pelvic lymphadenectomy. PMID- 12648869 TI - Prevalence of urinary incontinence in young and middle-aged women in an Italian urban area. AB - OBJECTIVE: To investigate by questionnaire the prevalence and characteristics of urinary incontinence (UI) in a female population aged between 18 and 49. STUDY DESIGN: Of 44,095 females aged 18-49.9 living in our urban area a sample of 10,000 subjects was sent a questionnaire and 3557 replied. The age-based analysis was performed on 2900 of these. RESULTS: The overall prevalence of UI was 20%. 11% had suffered from nocturnal enuresis before 18 years old, 47% of parous women noted a relationship between the beginning of UI and delivery. Among the incontinent women 83% lost urine with physical exertion, and 44% had urge incontinence, 18% used diapers, 20% had consulted a physician and half of these had been referred for medical, surgical or rehabilitative therapy, 50% reported past or present urinary infections, 8% complained also of faecal incontinence, 22% considered loss of urine embarrassing and humiliating but only 6% revealed frustration and 2% were obliged to stay at home for long periods of time. Of the incontinents 62% defined themselves anxious or nervous and 22% of these ascribed an increase of urinary leakage to anxiety or to nervousness, 9% reported use of antidepressant drugs, 85% had sexual intercourse and only 2% considered UI as an impediment to a satisfying sexual activity. CONCLUSIONS: UI is a common problem in young women. Urge incontinence seems to be prevalent in <30 aged women while stress incontinence occurs more frequently after this age. PMID- 12648870 TI - Pre-pubic TVT: an alternative to classic TVT in selected patients with urinary stress incontinence. AB - OBJECTIVES: To evaluate in a prospective open study a pre-pubic route of TVT for surgical treatment of female stress incontinence. STUDY DESIGN: Consecutively, 74 patients were operated using a pre-pubic TVT tape application. All women were suffering subjectively and objectively from female stress urinary incontinence. The mean post-operative follow-up time was 5 months (range 2-10 months). The pre- and post-operative evaluations were performed according to a standard protocol. RESULTS: According to the protocol, 60 patients (81%) were cured of their stress incontinence symptoms. Another 10 patients (13%) were improved. Four patients (6%) were considered failures. There were no significant intra- or post-operative complications. CONCLUSION: The short-term results of pre-pubic TVT are consonant with those of classic TVT. The risks of intra-operative complications should be reduced by the pre-pubic route. If the long-term results of pre-pubic TVT are the same as those after classic TVT, then this surgical approach may be a tentative alternative in selected high-risk patients. PMID- 12648871 TI - Gastric carcinoma with metastasis to the placenta and amniotic fluid: case report and review of the literature. PMID- 12648873 TI - Acardiac twin: a misdiagnosed, mismanaged case. AB - Acardiac twins are usually diagnosed during the antenatal period and almost invariably cause complications to the other fetus. In this case, an acardiac twin pregnancy was misdiagnosed antenatally as a degenerated myoma but ended with delivery of a healthy baby. PMID- 12648872 TI - Detection of fetal DNA in the peritoneal cavity during pregnancy. AB - There is increasing evidence that fetal cells are commonly shed toward the cervix and in maternal circulation during pregnancy. In this study, a sample of peritoneal fluid was retrieved from a primigravida at 12 weeks' gestation undergoing urgent intervention for the torsion of an adnexal mass; the sample was then analysed by a polymerase chain reaction (PCR) assay using primers for X- and Y-chromosome specific sequences, and Y-derived sequences were identified. The course of pregnancy was then uneventful until term, when the patient delivered a male fetus, thus, supporting the hypothesis of a fetal origin for the Y-derived sequences detected in the peritoneal fluid. Further studies are required in order to confirm these findings and precisely define the origin of these sequences; however, this report seems to provide further evidence of the spreading of fetal cells during gestation and addresses relevant issues as to the possibility of collecting these cells by culdocentesis and intraperitoneal lavage for prenatal diagnosis. PMID- 12648874 TI - Mediterranean spotted fever during pregnancy: case presentation and literature review. AB - Mediterranean spotted fever (MSF) is caused by Rickettsia conorii, an obligate intracellular parasite of eukaryotic cells. Although, usually this disease has a benign course, a rapidly fatal outcome can occur even in young healthy adults. We describe a case of a 40-year-old Bedouin woman gravida 11, para 10, who was admitted at 36 weeks gestation with this rickettsial disease. During pregnancy, the treatment of choice for Mediterranean spotted fever is chloramphenicol, but it seems that Azithromycin could be another possible option. PMID- 12648875 TI - Ovarian psammocarcinoma with peritoneal implants. AB - A case of ovarian psammocarcinomas with omental and peritoneal implants in a 48 year-old woman was treated with total hysterectomy, bilateral oophorectomy and omentectomy. Two years later there was no sign of recurrent disease. PMID- 12648876 TI - Enterobius vermicularis infection of female genital tract: a report of three cases and review of literature. PMID- 12648880 TI - P53 expression in spontaneous and estradiol-induced endometrial hyperplasia during menopause. AB - OBJECTIVE: To determine the percentage of endometrial hyperplasia positive for p53 expression in both spontaneously occurring cases or following the use of unopposed estradiol. METHODS: Fifty-four postmenopausal patients with endometrial hyperplasia diagnosed by endometrial biopsy and hysteroscopy were recruited to this study. Thirty-three patients had used unopposed estradiol for periods of time from 1 to 3 years. P53 expression was detected in paraffin-embedded endometrial specimens by immunohistochemical methods. RESULTS: The percentage of endometrial hyperplasia positive for p53 expression was significantly greater in spontaneously occurring hyperplasia than in cases induced by the unopposed use of estradiol. CONCLUSION: Endometrial hyperplasia caused by the unopposed use of estradiol during menopause probably harbors fewer genomic errors than those cases occurring spontaneously. PMID- 12648881 TI - Plasma and urinary sex hormones are differently related to lipids in healthy postmenopausal women. AB - OBJECTIVE: Endogenous sex hormones can be measured in plasma and urine. We determined the extent to which these two methods provide different information on hormonal status by relating them to lipid profile in postmenopausal women. METHODS: Thirty healthy postmenopausal women collected one 24-h urine sample and a blood sample was taken. Urinary estrone (UE), plasma estrone (PE) and serum lipids were measured. Sex hormone levels were measured with specific radioimmunoassays. Linear regression analysis was used to determine associations between estrone levels and lipids. Results are presented as beta-coefficients in mmol/l per standard deviation (SD) of endogenous estrone levels, adjusted for body mass index (BMI) and smoking (95% confidence interval). A stratified analysis for obese (BMI> or =27 kg/m(2)) versus lean women was performed. RESULTS: Mean levels of endogenous sex hormones were (SD): PE, 90.1 pmol/l (37.3); and UE, 7757 pmol/24 h (2659). PE showed significant associations with HDL-cholesterol (0.18 mmol/l, 95% CI: 0.06; 0.30), triglycerides (-0.25 mmol/l, 95% CI: -0.49; -0.009) and very-low-density-lipoprotein (VLDL-cholesterol) (-0.11 mmol/l, 95% CI: -0.22; -0.003), but not with total and low-density-lipoprotein (LDL-cholesterol). UE was inversely associated with total (-0.41 mmol/l, 95% CI: 0.85; 0.02) and LDL-cholesterols (-0.42 mmol/l, 95% CI: -0.83; -0.005), but not with HDL-cholesterol, triglycerides and VLDL-cholesterol. All associations appeared to be stronger in lean women. CONCLUSION: Both plasma and UE levels appear to be associated to serum lipids in healthy postmenopausal women. However, this relation appears to be different for estrone levels in plasma and urine. Depending on the research question, either blood samples or urine samples may be preferred. PMID- 12648882 TI - Adverse effects of a SERM (Levormeloxifene). Safety parameters and bone mineral density 12 months after treatment withdrawal. AB - OBJECTIVE: Levormeloxifene is a selective estrogen receptor modulator (SERM). The development of the drug was discontinued due to intolerable adverse effects. This paper follow-up on the adverse events in a group of 234 women that was followed for 12 months without treatment after 12 months of treatment with levormeloxifene. METHODS: Adverse events were recorded at all clinical visits. The double-layer thickness of the uterine endometrium was determined by transvaginal ultrasonography. Endometrial biopsies were obtained by pipelle. The biopsies taken at the entrance to the follow-up phase were taken under hysteroscopy-guidance. Bone mineral density of the total body, lumbar spine (L1 L4), hip and forearm was measured by dual-energy X-ray absorptiometry. RESULTS: The most prominent adverse event was increased endometrial thickness over the pre defined threshold of 8 mm. No cases of proliferative endometrium were reported. Following withdrawal of treatment the mean endometrial thickness approached baseline levels in a dose dependent manner. Hysteroscopic examinations showed that levormeloxifene was related to increased incidence of edema, vascularization and cysticity. In the levormeloxifene groups, a total of eight women had utero vaginal prolapse and five women reported urinary incontinence (including worsening of a previously existing condition). Bone density in the spine and hip approached baseline levels during the 12 months of follow-up without treatment. CONCLUSION: Endometrial thickening, seen in association with the use of some SERM's, may lead to harmful adverse effects more than 12 months after treatment is initiated. Levormeloxifene prevents the postmenopausal bone loss, but the lowest effective dose is unknown. PMID- 12648883 TI - Compliance with hormone replacement therapy in Thai women. AB - A retrospective study was performed to determine the compliance of hormone replacement therapy (HRT), the characteristics of women who continue to use HRT and the reasons for discontinuation. This study comprised of 821 postmenopausal women who attended the menopause clinic between January 1993 and December 1997. A total of 613 women (74.67%) were considered to be good compliant users. They had been on HRT for at least 1 year. Two hundred and eight women (25.33%) discontinued the treatment, with an average of 5.1 months duration of use. The younger age group at time of consultation and at menopause and those with previous hysterectomy were factors associated with higher degree of compliance. Its compliance decreased with age and the duration of menopause. The common reasons in the women's decision to discontinue HRT were bleeding episodes (23.08%), undesirable side effects (15.05%) and on physicians' advice (13.46%), respectively. Compliance with HRT appears to be similar to that reported in other specialist centres. Factors affecting compliance were age, age at menopause, duration of menopause, and hysterectomy. Irregular vaginal bleeding was the major reason for discontinuation. PMID- 12648884 TI - A prospective, randomized, placebo-controlled study of the dose effect of oral oestradiol on menopausal symptoms, psychological well being, and quality of life in postmenopausal Chinese women. AB - OBJECTIVES: Hypoestrogenism occurring in association with the menopause may result in the development of vasomotor symptoms and it may also have a detrimental effect on psychological well being and quality of life (QOL). The aims of this study were to measure menopausal symptoms, mood and QOL in postmenopausal Chinese women and to assess the effect of different doses of oestrogen on these outcome indicators. METHODS: A prospective, randomized, placebo-controlled study of the effect of 1 and 2 mg oestradiol on menopausal symptoms, anxiety and depressive symptoms, and QOL in 152 postmenopausal women over a 12 month study period. Menopausal symptoms were measured using a modified Kupperman's scale. Anxiety and depressive symptoms and QOL were measured using the Hospital Anxiety and Depression Scales and a modification of the World Health Organization Quality of Life questionnaire, respectively. RESULTS: Baseline scoring of vasomotor symptoms in our population was low whilst QOL scoring was relatively high. Over 12 months, after adjustment for differences in baseline scoring, there was a significant reduction in menopausal symptom scores in the 2 mg oestradiol group compared with placebo but not in the 1 mg group. There were no statistically significant changes in levels of anxiety and depression or QOL in either the 1 or the 2 mg group compared with placebo. CONCLUSIONS: These results suggest that relatively few Chinese women will be expected to benefit from hormone replacement in terms of either QOL or mood. In addition, the overall benefit of treatment for vasomotor symptoms will be less for a given number of Chinese women than for Caucasians. Therefore, when considering the reasons for prescribing hormone replacement therapy in this population, protection against osteoporosis will for most women be the prime consideration. PMID- 12648885 TI - Quantitative ultrasound of the calcaneus in Arabian women: relation to anthropometric and lifestyle factors. AB - OBJECTIVES: To determine factors influencing quantitative ultrasound (QUS) parameters of the calcaneus in a population-based sample of United Arab Emirates (UAE) women, and to compare QUS parameters of the calcaneus for healthy young UAE women with the manufacturer's reference ranges for other populations. METHODS: All subjects completed a questionnaire on reproductive and life style factors. Height and weight were measured, and body composition was determined by bioelectric impedence. Estimated bone mineral density (BMD), Speed of sound (SOS), broadband ultrasound attenuation (BUA) and quantitative ultrasound index (QUI) of the right calcaneus were determined by Sahara ultrasound. RESULTS: In premenopausal women (n=330), age, weight, body mass index (BMI), lean weight, fat weight, education, age at menarche, and number of pregnancies, correlated significantly with QUS parameters. Multiple regression analysis showed that age at menarche, number of pregnancies, and BMI, were the best predictors of QUS parameters although these factors explained only small amounts of the variance (R(2)=0.05). In postmenopausal women (n=81), age, BMI and physical activity were the best predictors of BUA (R(2)=0.35), SOS (R(2)=0.39), and QUI (R(2)=0.43). Mean estimated BMD, QUI and SOS for healthy young UAE women were significantly lower than the manufacturer's reference ranges for U.S. Caucasian, European Caucasian, and Chinese Asian healthy young women of the same age range (P<0.001 for all comparisons). Mean BUA was not significantly different, however. CONCLUSIONS: Menopausal status, age, BMI and physical activity are strong predictors of QUS parameters of the calcaneus in Arabian women. Healthy young Arabian women have lower estimated calcaneal BMD compared with the manufacturer's reference ranges for other populations. PMID- 12648886 TI - Symptom experience in Australian men and women in midlife. AB - OBJECTIVE: To compare the experience of vasomotor, psychological and somatic symptoms in Australian men and women in midlife, to investigate whether symptoms often attributed to endocrine changes in midlife are associated with health and psychological well-being, and to evaluate their relationship to menopausal status in women. METHODS: A cross-sectional, correlational, population-based study based on self-report questionnaires. Participants comprised 451 men and 766 women, aged between 39 and 65, from urban and rural South Australia, who responded to invitations to participate, or who volunteered to participate following media releases. Outcome measures used comprised a 47-item symptom checklist of current presence and severity of vasomotor, somatic and psychological symptoms experienced by men and women during midlife, and measures of health and psychological well-being. RESULTS: The majority of men and women reported that they were not 'bothered' by vasomotor, psychological and somatic symptoms. For those symptoms in which men and women differed significantly, women generally reported being more 'bothered' than men, although the pattern of association between symptoms and measures of health and psychological well-being was the same for both men and women. Items from all three symptom clusters were independently related to menopausal status. CONCLUSIONS: Health and psychological well-being play a role in the genesis of symptoms experienced by men and women in midlife. Both men and women experience similar symptoms, although women are more distressed by them signifying support for a menopausal syndrome in women. The finding of an independent relationship between menopausal status and psychological and somatic symptoms, in addition to the vasomotor symptoms, contradicts the narrow-estrogen hypothesis of climacteric symptoms. PMID- 12648887 TI - Performance and acceptability of intrauterine release of levonorgestrel with a miniature delivery system for hormonal substitution therapy, contraception and treatment in peri and postmenopausal women. AB - OBJECTIVE: To evaluate the performance and acceptability of a novel intrauterine drug delivery system, FibroPlant-levonorgestrel (LNG), delivering approximately 14 microg per day. SUBJECT AND DESIGNS: A 1-year prospective clinical trial in 141 peri and postmenopausal women, including women with heavy or postmenopausal bleeding and women needing contraception. The majority received percutaneous 17beta-estradiol (Oestrogel), 1.5 mg daily. Clinical results and ultrasonographic effects were evaluated. RESULTS: Eighty-three insertions were done in perimenopausal women and 58 in postmenopausal women followed-up for 8-38 months. Fifty-two perimenopausal (64%) and virtually 100% of the postmenopausal women developed amenorrhoea, with occasional slight spotting. Eleven women with heavy bleeding, five of them with single or multiple intramural and subserosal fibroids of 3-6 cm or more, were all successfully treated, except one. There were no pregnancies. CONCLUSION: This study of 1432 women-months of use suggests that the frameless FibroPlant-LNG IUS is safe, well tolerated and effective in suppressing the endometrium during EST. The fact that the IUS also acts as a contraceptive, and significantly reduces menstrual bleeding, as demonstrated in earlier studies, is of added importance. PMID- 12648889 TI - The communicative use of pointing in autism: developmental profile and factors related to change. AB - PURPOSE: To describe different longitudinal profiles in communicative abilities and symptoms severity in early autism. METHODS: Five children with autism, aged from 3;3 to 4;10 years at baseline, were tested for production and comprehension of imperative and declarative pointing at about 4-month intervals. Concurrently with these sessions, children were evaluated in terms of cognitive and communicative abilities, and symptoms severity. RESULTS: Four subjects showed a mild to severe retardation in communicative and linguistic abilities. For production, all children exhibited the imperative pointing and only one the declarative pointing. For comprehension, two subjects showed the same profile as in production ('only imperative' and 'first imperative-later declarative', respectively). One child did not show any clear comprehension of the pointing gestures produced by the experimenter, and one child was able to understand both pointing in the same session. Childhood autism rating scale (CARS) global scores tended to decrease across sessions for all subjects and different individual profiles were identified. DISCUSSION: Declarative or experience-sharing pointing emerged later in one child only; it remained absent in four children as production, and in two children as comprehension. A preliminary conclusion based on CARS rating, is that autism involves a symptomatology that may decrease across time even if children differ in the decreasing profile relative to specific scores. PMID- 12648888 TI - Four-year outcome in non-compliant schizophrenia patients treated with or without home-based ambulatory outpatient care. AB - Non-compliance in neuroleptic maintenance treatment is a major concern in schizophrenia. Home-based outpatient care has been shown both to improve medication compliance and reduce relapse frequency. We analysed the need for hospitalisation, levels of functioning and mortality rate during the de institutionalisation process in 41 schizophrenia patients with repeated hospitalisations and prolonged history of non-compliance. Eighteen of the patients received ambulatory outpatient care (AOC) after discharge. This treatment procedure focuses on enduring neuroleptic maintenance treatment. One of the hospital nurses takes care of home visits every 2-4 weeks. In the 4-year follow-up, half of the patients in the AOC group did not need hospitalisation at all and the number of days of hospitalisation in the whole group diminished by almost four-fifths compared with the previous 4 years. In the non-AOC group, there was a more limited decrease in the number of days of hospitalisation during the corresponding follow-up period. The mortality rates showed a slight tendency towards a better outcome in the AOC group. There was no change in the levels of functioning in the AOC group. This treatment can be carried out with limited resources. It clearly reduces the need for hospitalisation in a subgroup of schizophrenia patients having problems with compliance and recurrent relapse. The effectiveness of AOC on the mortality rates of schizophrenia patients needs further examination. PMID- 12648890 TI - The Maudsley bipolar disorder project. Clinical characteristics of bipolar disorder I in a Catchment area treatment sample. AB - The clinical characteristics of bipolar I disorder (BD1) have prognostic and therapeutic importance. The aim of this study was to examine the effect of demographic and clinical variables on the course of BD1. We reviewed the case notes of all BD1 patients (n = 63) receiving treatment in a London psychiatric service during a 1-month period. Depressive and manic onsets were equally likely without any gender difference. The earlier the age of onset, the more likely it was for patients to experience psychotic features. Only depressive onsets predicted a higher number of episodes of the same polarity. Male gender and substance abuse were associated with younger age at first presentation, while women with co-morbid substance abuse had more manic episodes. Male patients were more likely than females to be unemployed or single. PMID- 12648892 TI - Cholesterol concentrations in violent and non-violent women suicide attempters. AB - The aim of this study was to evaluate whether women with a history of violent suicide attempts have lower serum cholesterol concentrations than those who attempted suicide by non-violent methods. Our retrospective study used a case control design to compare serum total cholesterol concentration, hematocrit, red blood cell count and body mass index (BMI) in women with a history of violent (n = 19) or non-violent (n = 51) suicide attempts and of non-suicidal controls (n = 70) matched by diagnosis and age. Analysis of covariance (ANCOVA) with age as the covariate was used to analyze differences in cholesterol levels in groups according to violence. Violence was found to be a significant factor (P = 0.016). Using the Scheffe test, a significant difference (P = 0.011) was revealed between the group of violent and non-violent suicide attempters and between the violent suicide attempters and the control group. Patients with a violent suicidal attempt have significantly lower cholesterol levels than patients with non violent attempts and the control subjects. Our findings suggest that suicide attempts should not be considered a homogeneous group. They are consistent with the theory that low levels of cholesterol are associated with increased tendency for impulsive behavior and aggression and contribute to a more violent pattern of suicidal behavior. PMID- 12648891 TI - Introducing a new recruitment approach to sample collection for genetic association studies in opioid dependence. AB - OBJECTIVE: In a modified case-control association study we tested the assumption that two polymorphisms (A(118)G in exon 1 and IVS2+31 in intron 2) of the human mu-opioid receptor gene (OPRM1) confer susceptibility to opioid dependence. METHODS: In contrast to classical case-control studies both groups, opioid dependent cases and non-opioid dependent controls were recruited from individuals who have had access to drugs including opioids and who had been sentenced for violation of the "Dangerous Drugs Act" in Germany. RESULTS: For the two allelic variants of OPRM1 under study we did not find evidence for association with opioid dependence. CONCLUSIONS ;Despite absence of association we think that this recruitment approach introduced here, is useful since it putatively offers a more adequate matching for case-control association studies of opioid dependent individuals. PMID- 12648893 TI - Zuclopenthixol-acetate treatment in catatonic patients: the implication of iron metabolism. AB - There is some evidence of ferropenia correlating with neuroleptic malignant syndrome and catatonic symptoms. The aim of this prospective and naturalistic study was to investigate the implications of ferremia in patients undergoing an intramuscular injection treatment of Zuclopenthixol-acetate in Visceolo. We recruited 59 catatonic patients (33 females). Age, sex, psychiatric and somatic diagnoses, body mass index (BMI), dosage and duration of Zuclopenthixol-acetate medication and the timing of the changeover from intramuscular to oral prescription, the single dosage of Clopenthixol if initially coadministered, incidence, onset and duration of transient benign hyperthermia, iron, ferritin, transferrin and saturation values, and white and red blood cell counts as well as liver function and electrolytes were registered. A transient and benign hyperthermic reaction (mean degrees: 37.5 + 0.3 degrees C) lasting for an average of 3.0 + 1.9 d was shown by 72.9% patients (N = 43, 22 females), during a mean treatment period of 5.8 + 3.1 d. These patients were medicated with significant different mean doses of Zuclopenthixol-acetate and compared to the patients with normal body temperature (ANOVA P < 0.01). The duration of Zuclopenthixol-acetate application did not vary between these patients groups. Furthermore, significant differences of iron (59.5 + 30.6 micromol/dl vs. 87.8 + 40.8 micromol/dl; ANOVA P < 0.006) and transferrin saturation values (18.3 + 10.4% vs. 27.2 + 17.0%; ANOVA P < 0.02) were found. Ferritin and transferrin were not implicated in the episode of hyperthermia. Diagnoses, sex, white and red blood cell counts also did not vary between these groups. Our findings indicate a possible involvement of ferropenia in catatonic patients, regardless of the diagnoses, and in the development of benign transient hyperthermia, also known as drug fever. PMID- 12648894 TI - Three case reports of secondary mania: evidence supporting a right frontotemporal locus. PMID- 12648895 TI - The clinical use of milnacipran for depression. AB - We examined the relation between dosage and efficacy, and the predictors of response to milnacipran. There was no difference between 50 and 100 mg dose. However, the 100 mg dose had a faster onset of action than the 50 mg dose. An age and an episode have been predictors of milnacipran. PMID- 12648896 TI - Social characteristics of seasonal affective disorder patients: comparison with suicide attempters with non-seasonal major depression and other mood disorder patients. AB - Although it is evident from numerous studies that patients with mood disorders generally have a deficient social functioning and a weak social network, little is known about these aspects of seasonal affective disorder (SAD) patients. We studied the social situation, the social network and the social functioning of SAD (n = 20) patients in comparison with matched suicide attempters (SA) with non seasonal major depression, and with findings from other major depressive disorder (MDD) studies and community samples. The social situation and the clinical background of both the SAD and the SA groups were almost similar and the social networks were equally disadvantageous and weaker than those observed in some community/healthy populations. Furthermore, the data on global functioning and social adjustment of the SAD group were well comparable to those of other MDD patients and significantly worse than that of a community sample. Thus, the results indicate a considerable social impairment in SAD. PMID- 12648897 TI - Which neuroleptic would psychiatrists take for themselves or their relatives? AB - OBJECTIVE: Psychiatrists should be asked which neuroleptic they would prefer for themselves and their relatives in the case of first manifestation of schizophrenia. METHOD: Questionnaires were answered by 66 psychiatrists from 13 different sites in Baden-Wuerttemberg (South West Germany). Only those were included who had treated more than 50 patients with schizophrenia within the last 5 years (n = 54). RESULTS: The psychiatrists were experienced with conventional and most of the atypical agents. Fifty-one point nine percent would take olanzapine as first line treatment, 20.4% risperidone, 13.0% quetiapine, 9.3% amisulpride, 3.7% haloperidol and 1.9% perazine. In four cases, different substances were preferred for oneself and relatives. CONCLUSION: Most psychiatrists would not take conventional neuroleptics, though 70% of prescriptions for schizophrenic patients have been conventional neuroleptics in Germany in 2000. The preferences among atypicals correspond well with the current prescription practice in Germany. PMID- 12648898 TI - Clinically relevant reduction of lamotrigine concentrations by carbamazepine. PMID- 12648899 TI - Paroxetine-induced enuresis. PMID- 12648902 TI - The natural history of untreated skull base meningiomas. AB - BACKGROUND: This study was undertaken to determine the natural history of untreated skull base meningiomas. Although there are reports on the natural history of meningiomas, most series contain only a few cases of meningiomas involving the skull base. Natural history information is important when recommending treatment and evaluating results. METHODS: The case records and imaging studies of 40 patients with skull base meningiomas from a single practice were reviewed. These patients either did not receive treatment or there was a long delay from the onset of symptoms to the initiation of treatment. RESULTS: Skull base meningiomas may be very indolent tumors. Patients with these tumors have lived long, comfortable, and relatively productive lives without treatment or with delayed treatment. Impairment may be much less than imaging studies would suggest. CONCLUSION: The authors are not advocating that skull base meningiomas not receive treatment. However, their unique experience with this group of patients provides convincing evidence that the indolent behavior of some skull base meningiomas must be considered when recommending various surgical and radiation procedures and interpreting their results in relationship to the functional outcome of the patient. PMID- 12648904 TI - Demonstration of neurovascular compression in trigeminal neuralgia and hemifacial spasm with magnetic resonance imaging: comparison with surgical findings in 60 consecutive cases. AB - BACKGROUND: Until recently, it has been impossible to demonstrate vascular compression at the root entry or exit zone (REZ) of the trigeminal nerve and facial nerve in patients with trigeminal neuralgia (TN) and hemifacial spasm (HFS) preoperatively, although surgical findings have revealed apparent neurovascular compression and its correction has resulted in a good outcome in most cases. Revealing the anatomic correlation between nerves and vessels at the REZ preoperatively would be useful to predict operative findings. METHODS: To assess whether the vascular contact of the nerve at the REZ could be demonstrated preoperatively, high-resolution magnetic resonance tomographic angiography (MRTA) was performed in 21 patients with TN and 39 with HFS. Neuroradiological findings were compared with the operative findings in all patients. Contralateral asymptomatic nerves were evaluated as a control. RESULTS: MRTA correctly identified offending vessels in 14 (67%) of the 21 TN and 34 (87%) of the 39 HFS patients. Failure to identify neurovascular contact was noted in the cases with compression by veins or small arteries, thickened arachnoid, or distal compression. Neurovascular contact was also observed in 15% of the asymptomatic nerves. The deformity of the nerve seemed to be a more important factor for determining operative indication. CONCLUSIONS: MRTA could demonstrate offending vessels in TN and HFS at a high rate and was useful to predict operative findings. MRTA gave supportive evidence of surgical indications in patients with TN and HFS, although attention should be paid to the fact that MRTA did not necessarily detect all of the offending vessels. PMID- 12648906 TI - Microsurgical excision of symptomatic sacral perineurial cysts: a study of 15 cases. AB - BACKGROUND: The widespread use of magnetic resonance imaging (MRI), now the first line investigation for back and leg pain, reveals cystic sacral lesions more often than myelography did in the past. There is agreement that symptomatic perineurial sacral cysts should be treated surgically. However, it is still debated whether the preference should be given to the curative option, consisting of excision of the cyst with duraplasty, or to drainage of the cyst to relieve symptoms. In this retrospective study the efficacy of microsurgical cyst resection with duraplasty is evaluated. METHODS: In 15 patients presenting with pain and neurologic deficits, myelography and/or MRI detected sacral cysts. The clinical features suggested that the space-occupying lesions caused the disturbances. Microsurgical excision of the cyst along with duraplasty or plication of the cyst wall was performed in all the cases. Postoperative care included bed rest and CSF drainage for several days. RESULTS: In 13 out of 15 patients the preoperative radicular pain disappeared after surgery. The 2 patients with motor deficits and the 6 patients with bladder dysfunction recovered completely. In all except 1 of the 10 patients complaining of sensory disturbances a significant improvement was achieved. No complications were observed. CONCLUSION: Microsurgical excision of the cyst combined with duraplasty or plication of the cyst wall is an effective and safe treatment of symptomatic sacral cysts and, in the view of the authors, the method of choice. PMID- 12648909 TI - Surgery for posterior communicating artery aneurysms. AB - The most common site of internal carotid artery (ICA) aneurysms is at the posterior communicating artery (PComA) junction. Although ICA aneurysm surgery carries substantial risk, the ultimate outcome of patients with PComA aneurysms is better than those with aneurysms in other locations. The rate of successful surgical outcome increases when precise knowledge of the anatomy and understanding of the hazards associated with aneurysms in this region are combined with a well organized plan for the surgical approach. This article will describe details of the preoperative considerations, operative techniques, and surgical results for aneurysms arising at the ICA-PComA junction based on the 25 year personal experience of the senior author, comprising a total of 424 surgical cases. PMID- 12648910 TI - Rupture of middle cerebral artery aneurysm in a neonate: case report and review of the literature. AB - Intracranial hemorrhage because of rupture of a cerebral aneurysm is extremely rare in the neonatal period. Delayed diagnosis contributes to high mortality and morbidity. The authors report an extremely rare case of a middle cerebral artery aneurysm diagnosed and treated shortly after birth. Extensive review of the literature is presented. The patient died 4 years after surgery. PMID- 12648913 TI - Pineal region lesion masquerading choroid plexus papilloma: case report. AB - BACKGROUND: Choroid plexus papillomas (CPPs) are rare intracranial neoplasms, especially in the third ventricle. We report a patient with a posterior third ventricular CPP extending into the pineal that radiographically and clinically presented as a pineal region lesion. CASE DESCRIPTION: In a 51-year-old female with headache and upward gaze impairment radiological examination showed a mass in the pineal region obstructing the aqueduct of Sylvius and causing hydrocephalus. After ventriculoperitoneal shunting the tumor was approached through the infratentorial-supracerebellar approach and pathological examination revealed a typical CPP. CONCLUSIONS: This case represents an unusual presentation of an intracranial CPP. This entity should be considered an extremely rare cause of a lesion in the pineal region. PMID- 12648911 TI - Congenital cavernous angioma of the temporal bone: case report. AB - BACKGROUND: Congenital cavernous angioma is a very rare tumor that occurs in young adolescents. We describe a rare case of congenital cavernous angioma of the temporal bone in a neonate. Ultrasonography (US) and dynamic magnetic resonance imaging (MRI) were helpful in its diagnosis. CASE DESCRIPTION: US and MRI of the fetus in the 32nd week of gestation revealed a large extra-axial tumor with intra and extracranial extension. After the birth, US and dynamic MRI studies were performed in addition to conventional CT and MRI. These findings were compatible with cavernous angioma. The mass was totally removed on the fourth day of life and was found at surgery to originate from the skull. Histopathological diagnosis was cavernous angioma. The usefulness of US and dynamic MRI are also discussed. CONCLUSION: US and dynamic MRI are useful, less invasive techniques for diagnosing this rare type of tumor. They provide useful information to differentiate calvarial mass lesions in neonates. PMID- 12648920 TI - Insurance and the depersonalization of medicine. PMID- 12648917 TI - Phrenic nerve stimulation for diaphragm pacing with a spinal cord stimulator: technical note. AB - BACKGROUND: Diaphragm pacing with electrical stimulation of the phrenic nerve is an established treatment for central hypoventilation syndrome. The device, however, is not readily available, at least in Japan. We used the spinal cord stimulator for pain control for phrenic nerve stimulation. The purpose of this study is to evaluate the efficacy and feasibility of phrenic pacing with the compromise method. METHODS: We implanted a stimulator for spinal cord stimulation (Itrel 3 or X-trel, Medtronic, MN) in 4 patients with chronic hypoventilation because of brainstem dysfunction of various origins. The stimulation electrode was placed along the right phrenic nerve in the neck, and the device was implanted in the anterior chest. We used the cyclic mode, and set the parameters at 1 second ramp up, 2 seconds on, 3 seconds off. The pulse width and the frequency were set at 150 microsec and 21 Hz, respectively. The amplitude of the output was adjusted to obtain sufficient tidal volume and to maintain PaCO(2) at around 40 mm Hg. RESULTS: During the follow-up period from 6 to 19 months (mean 8.3 months), stable and sufficient ventilation were observed in all patients without complications. One patient with sleep apnea syndrome used the device only at night and became free from a respirator. Three patients who were completely respirator-dependent became ambulatory during the daytime. CONCLUSION: Though longer follow-up is necessary, diaphragm pacing by stimulation of the phrenic nerve with the spinal cord stimulator is feasible for a treatment of central hypoventilation syndrome. PMID- 12648921 TI - Treat the patient not the disease. PMID- 12648922 TI - Meningiomas--the future: is 2020 here? PMID- 12648925 TI - Electron ionization (EI) mass spectra of exo-endo double-bond isomers of polycyano "push-pull" pentadienes derived from cycloalkylidene malonic acid derivatives. AB - The title compounds, which exist in solutions as mixtures of exo/endo double bond isomers due to the "push-pull" effect of the electron-donating and electron withdrawing substituents, were studied by mass-spectrometric methods. Their fragmentation routes under electron impact were established and confirmed by metastable ion analysis and accurate mass measurements. The results demonstrated that the relative amounts of exo/endo isomeric molecular ions are in close agreement with the isomeric ratios observed in solutions by the NMR, although the mass spectra of the tetracyano derivatives indicated a small fraction of molecular ions existing in the endo form, which could not be detected in solution by the NMR methods. PMID- 12648924 TI - Ketalization of phosphonium ions by 1,4-dioxane: selective detection of the chemical warfare agent simulant DMMP in mixtures using ion/molecule reactions. AB - Phosphonium ions CH(3)P(O)OCH(3)(+) (93 Th) and CH(3)OP(O)OCH(3)(+) (109 Th) react with 1,4-dioxane to form unique cyclic ketalization products, 1,3,2 dioxaphospholanium ions. By contrast, a variety of other types of ions having multiple bonds, including the acylium ions CH(3)CO(+) (43 Th), CH(3)OCO(+) (59 Th), (CH(3))(2)NCO(+) (72 Th), and PhCO(+) (105 Th), the iminium ion H(2)C[double bond]NHC(2)H(5)(+) (58 Th) and the carbosulfonium ion H(2)C[double bond]SC(2)H(5)(+) (75 Th) do not react with 1,4-dioxane under the same conditions. The characteristic ketalization reaction can also be observed when CH(3)P(OH)(OCH(3))(2)(+), viz. protonated dimethyl methylphosphonate (DMMP), collides with 1,4-dioxane, as a result of fragmentation to yield the reactive phosphonium ion CH(3)P(O)OCH(3)(+) (93 Th). This novel ion/molecule reaction is highly selective to phosphonium ions and can be applied to identify DMMP selectively in the presence of ketone, ester, and amide compounds using a neutral gain MS/MS scan. This method of DMMP analysis can be applied to aqueous solutions using electrospray ionization; it shows a detection limit in the low ppb range and a linear response over the range 10 to 500 ppb. PMID- 12648923 TI - Multiplex inhibitor screening and kinetic constant determinations for yeast hexokinase using mass spectrometry based assays. AB - An electrospray ionization mass spectrometry based assay was developed for kinetic measurements and inhibitor screening of yeast hexokinase. There is considerable discrepancy in the literature as to the accuracy of kinetic data obtained for hexokinase. In the assay described herein, the product, glucose 6 phosphate was directly monitored by ion trap mass spectrometry and quantified using an internal standard, 2 deoxy-glucose 6-phosphate. The kinetic parameters, K(M) and V(max) for the two substrates were determined without using a coupling enzyme as is normally employed in the traditional spectrophotometric assay for systems lacking a chromophore. In addition, hexokinase was successfully immobilized onto an amino-link gel, and a mock library was screened against the immobilized enzyme for the identification of possible inhibitors. After comparing the mass spectra of the library before and after incubation, trehalose 6 phosphate, ADP, and oxidized glutathione were differentiated from other weak or non-inhibitors. Inhibition behavior of ADP with respect to ATP was further evaluated with the ESI-MS assay and the value of K(i) was determined. This ESI-MS assay was demonstrated to be both accurate and precise for determining kinetic constants and for identifying enzyme inhibitors. PMID- 12648926 TI - Comparative studies of poly(dimethyl siloxanes) using automated GPC-MALDI-TOF MS and on-line GPC-ESI-TOF MS. AB - In this study we compare on-line gel permeation chromatography (GPC) electrospray ionization (ESI) time-of-flight (TOF) mass spectrometry (MS) to automated GPC matrix assisted laser desorption ionization (MALDI) TOF MS for poly (dimethylsiloxane) (PDMS) analysis. Average mass values for a hydroxyl-terminated PDMS (OH-PDMS) sample were obtained and compared to traditional GPC that was calibrated with narrow polystyrene standards, by direct ESI and MALDI MS analysis, by a summation of mass spectra of all GPC fractions, and also by the recalibration method determined by both mass spectrometric methods. Quantitatively, the difference noted here between these hyphenated techniques is that GPC-ESI-TOF MS effectively reports the low-mass oligomers and underestimates the high-mass oligomers, while GPC-MALDI-TOF MS effectively reports the high-mass oligomers and underestimates the low-mass oligomers. In the GPC-ESI-TOF MS experiments, ion current suppression was observed in the high molecular weight region. The suppression effect was confirmed by repeatable sample runs and by injecting different PDMS samples. Higher chromatographic resolution was observed for GPC-ESI-TOF MS compared to GPC-MALDI-TOF MS. In fact, truly mono-disperse oligomers were observed in the low molecular weight range from GPC-ESI MS experiments. PMID- 12648927 TI - Mass spectrometric study of the effects of hydrophobic surface chemistry and morphology on the digestion of surface-bound proteins. AB - Our previous work has demonstrated that reversed-phase chromatographic micro beads can be used to capture proteins from complex biological matrices and the surface-bound proteins can be enzymatically digested for protein identification by mass spectrometry (MS). Here we examine the peptides generated from digestion of proteins bound to various types of micro-bead surfaces in order to determine the effects of surface chemistry and surface morphology on the digestion process. Detailed examinations of site cleavages and sequence coverage are carried out for a tryptic digestion of cytochrome c adsorbed on reversed-phase polystyrene divinylbenzene (Poros R2 beads) versus C(18) bonded-phase silica beads. It is shown that although the surface does not completely hinder the digestion of cleavage sites of the protein, the digestion products are clearly different than those obtained from a solution digest. Specifically, a partial digestion results from surface digestion, resulting in a greater number of missed cleavages than a comparable solution digest. Subsequent comparisons of peptide mass maps generated from the digestion of various proteins on surfaces with altering chemistry (C(4), C(8), C(18), and R2 beads), or with different surface morphology, were performed. The results reveal that surface chemistry plays only a minor role in affecting the peptide mass maps, and surface morphology had no noticeable effects on the resulting peptide mass maps. It is also shown that the mass spectrometric detection method used to analyze the digested peptides can significantly influence the information content on cleavage sites and the extent of sequence coverage. The use of a combination of MALDI, LC/off-line MALDI, and LC/ESI MS is demonstrated to be crucial in revealing subtle changes in the peptide mass maps. PMID- 12648929 TI - Simultaneous or scanning data acquisition? A theoretical comparison relevant to inductively coupled plasma sector-field mass spectrometers. AB - The benefits of simultaneous multichannel detection over single-channel scanning detection are well established in analytical chemistry. Multichannel detection increases duty cycle, which leads to enhanced sensitivity, detection limits, and reduced analysis time. Also, multichannel detection used with either isotope ratio or internal-standard techniques provides a mechanism to reduce the effect of multiplicative or flicker noise prevalent in plasma sources. An additional benefit of simultaneous detection is superior analysis of short-lived transient signals. Presented here is a theoretical comparison between simultaneous/continuous multichannel acquisition and single-channel scanning acquisition. To conduct this comparison, reported sensitivity, single-channel precision, and background values for commercial inductively coupled plasma sector field mass spectrometers (ICP-SFMS) are used to generate theoretical figures of merit for both acquisition methods. Among the figures of merit that will be considered are detection limits, precision, and analysis time, particularly for multi-element or multi-isotope analysis. PMID- 12648928 TI - Study of protein modification by 4-hydroxy-2-nonenal and other short chain aldehydes analyzed by electrospray ionization tandem mass spectrometry. AB - A convenient way to study lipid oxidation products-modified proteins by means of suitable model systems has been investigated. As a model peptide, the oxidized B chain of insulin has been chemically modified by either 4-hydroxy-2-nonenal (HNE) or hexanal and the extent, sites, and structure of modifications were assessed by electrospray mass spectrometry. A reduction step, using either NaCNBH(3) or NaBH(4), was also studied to stabilize the alkylated compounds. From the data gathered, it appeared that NaCNBH(3), when added at the beginning of incubation, dramatically influenced the HNE-induced modifications in terms of the addition mechanism (Schiff base formation instead of Michael addition) but also of the amino acid residues modified (N-terminal amino acid instead of histidine residues). However, by reducing the HNE-adducted species at the end of the reaction with NaBH(4), the fragment ions obtained in the product ion scan experiments become more stable and thus, easier to interpret in terms of origin and mechanism involved. With regard to hexanal induced modifications, we have observed that hexanal addition under reductive conditions led to an extensive modification of the peptide backbone. Moreover, as confirmed by "in-source" collision followed by collision induced dissociation (CID) experiments on selected precursor ions (pseudo-MS(3) experiments), N,N-di-alkylations were first observed on the N-terminal residue and further on Lys(29) residue. On the other hand, compared to the native peptide, no significant changes in MS/MS fragmentation patterns (b and y ions series) were observed whatever the basic site modified by the aldehyde-addition. PMID- 12648930 TI - Classification of lactate dehydrogenase of different origin by liquid chromatography-mass spectrometry and multivariate analysis. AB - A method intended to serve as a multivariate quality control tool in the production of pharmaceutical proteins is presented. The method is based on multivariate analysis of peptide maps generated with liquid chromatography-mass spectrometry (LC-MS). Lactate dehydrogenase (LDH) from different species and tissues were used as model compounds in the study. The proteins were digested with Endoproteinase Lys-C before the LC-MS analysis. After data pretreatment of the peptide maps, successful classification of the LDHs were obtained by discriminant analysis with partial least squares regression and artificial neural networks. Further, principal component analysis was applied to visualize the relationships between the samples. PMID- 12648931 TI - Experimental and theoretical study of the gas-phase interaction between ionized nitrile sulfides and pyridine. AB - The gas-phase reactivity of ionized nitrile sulfides, R-C[triple bond]N(+)-S*, towards neutral pyridine was studied both experimentally (six sector hybrid mass spectrometer) and theoretically (density functional theory and Moller-Plesset ab initio calculations). An ionized sulfur atom transfer and a cycloaddition process respectively yielding ionized pyridine N-thioxide and a thiazolopyridinium cation were observed. Whereas the very efficient S*+ transfer reaction probably involves the intermediacy of several ion-molecule complexes, the thiazolopyridinium ion formation is likely to be initiated by an electrophilic attack of the R-C[triple bond]N(+)-S* ion on the nitrogen atom of pyridine; the resulting intermediate then undergo an intramolecular substitution of an alpha-hydrogen atom by the sulfur atom. PMID- 12648933 TI - On the comparison of different tests for identification of a compound from its mass spectrum. AB - It is shown that identification tests of different dimensions or dimensionless should not be evaluated (for their efficiency to identify molecules from their mass spectrum) by comparing the tests for a molecule itself (repeated measurements) with other molecules. This kind of tests must have similar dimensions (units). Another possibility is the comparison of tests on the basis of success of correct prediction for "unknown" molecules from a library of standards. PMID- 12648932 TI - Top down characterization of secreted proteins from Mycobacterium tuberculosis by electron capture dissociation mass spectrometry. AB - Secreted proteins of Mycobacterium tuberculosis are implicated in its disease pathogenesis and so are considered as potential diagnostic and vaccine candidates. The search for these has been slow, even though the entire genome sequence of M. tuberculosis is now available; of the 620 protein spots resolved by 2-D gel electrophoresis, 114 secreted proteins have been identified, but for only 13 has the primary structure been partly characterized. For comparison, in this top down mass spectrometry (MS) approach the secreted proteins were precipitated from cell culture filtrate, resuspended, and examined directly by electrospray ionization (ESI) Fourier transform MS. The ESI spectra of three precipitates showed 93, 535, and 369 molecular weight (M(r)) values, for a total of 689 different values. However, only approximately 10% of these values matched (+/-1 Da) the DNA predicted M(r) values, but these identifications were unreliable. Of nine molecular ions characterized by MS/MS, only one protein match was confirmed, and its isotopic molecular ions were overlapped by those of another protein. MS/MS identified a total of ten proteins by sequence tag search, of which three were unidentified previously. The low success of M(r) matching was due to unusually extensive posttranslational modifications, including loss of a signal sequence, loss of the N-terminal residue, proteolytic degradation, oxidation, and glycosylation. Although in eubacteria the latter is relatively rare, a 9 kDa protein showed 7 hexose attachments and two 20 kDa proteins each had 20 attachments. For MS/MS, electron capture dissociation was especially effective. PMID- 12648935 TI - Effects of modulation defects on Hadamard transform time-of-flight mass spectrometry (HT-TOFMS). AB - In any Hadamard multiplexing technique, discrepancies between the intended and the applied encoding sequences may reduce the intensity of real spectral features and create discrete, artificial signals. In our implementation of Hadamard transform time-of-flight mass spectrometry (HT-TOFMS), the encoding sequence is applied to the ion beam by means of an interleaved comb of wires (Bradbury Nielson gate), which shutters the ion beam on and off. By isolating and exaggerating individual skewing effects in simulating the HT-TOFMS process, we determined the nature of errors that arise from various defects. In particular, we find that the most damaging defects are: mismatched voltages between the wire sets and the acceleration voltage of the instrument, which cause positive and negative peaks throughout mass spectra; insufficient deflection voltage, which reduces the intensity of real peaks and causes negative peaks that are spread across the entire mass range; and voltage errors as the wire sets return from their deflection voltage to their transmission value, which yield significant reductions in peak intensities, create artificial peaks throughout mass spectra, and broaden real peaks by causing positive peaks to grow in the bins adjacent to them. Because the magnitude of the modulation defects grows as the applied modulation voltage is increased, Bradbury-Nielson gates with finer wire spacing, and hence stronger effective fields for a given applied voltage, were produced and installed. Operating at 10 to 15 V where errors in the electronics are essentially absent, the most finely spaced gate (100 microm) yielded signal-to noise ratios that were more than two times higher than those achieved with more widely spaced gates. As an alternative method for minimizing skewing effects, HT TOFMS data were post processed using an exact knowledge of the modulation defects. Nonbinary matrices that mimic the actual encoding process were built by measuring voltage versus time traces and then translating these traces to transmission versus time. Use of these matrices in the deconvolution step led to marked improvements in spectral resolution but require full knowledge of the encoding defects. PMID- 12648934 TI - Rapid and sensitive differentiation of anomers, linkage, and position isomers of disaccharides using High-Field Asymmetric Waveform Ion Mobility Spectrometry (FAIMS). AB - A challenging aspect of structural elucidation of carbohydrates is gaining unambiguous information for anomers, linkage, and position isomers. Such isomers with identical mass can't be easily distinguished in mass spectrometry and a separation step is required prior to mass spectrometry identification. In our laboratory, gas-phase separation and differentiation of anomers, linkage, and position isomers of disaccharides was achieved using High-Field Asymmetric Waveform Ion Mobility Spectrometry (FAIMS). The FAIMS method responds to changes in ion mobility at high field rather than absolute values of ion mobility, and was shown to provide efficient separation and identification of disaccharide isomers at high sensitivity. Separation of analyzed disaccharide isomers can be accomplished at low nM level in a matter of seconds without sample purification or fractionation. Capability for examining a large population of ionic species of disaccharides by this method allowed for correlating structural details of disaccharide isomers with their separation properties in FAIMS. Results for disaccharide isomers indicate that this method could be applied to a larger group of carbohydrates. PMID- 12648936 TI - Mimicking the way to autoimmunity: an evolving theory of sequence and structural homology. AB - Although the etiology of autoimmune diseases remains largely unknown, a prevailing theory concerns the infection-induced activation of self-reactive lymphocytes via the process of molecular mimicry. Here, we discuss the theory of molecular mimicry and its continued evolution from the initial basic considerations of sequence similarity to the current theories of structural homology. Such findings serve to further our understanding of T-cell receptor degeneracy and might one day provide a direct link between infection and autoimmune disease. PMID- 12648937 TI - Hypervirulence and pathogen fitness. AB - Historically, pathogenesis research has focused on the identification and characterization of virulence factors. More recently, 'anti-virulence' genes have been discovered. Mutations in these loci result in a hypervirulent phenotype, as measured by a lower lethal dose, a colonization advantage, reduced clearance or decreased survival time of the host. If these genes function to reduce pathogen virulence, why have they been retained? Multiple hypotheses have been offered to explain this phenomenon. PMID- 12648938 TI - The importance of being outer: consequences of the distinction between the outer and inner surfaces of flavivirus glycoprotein E. AB - Glycoprotein E of West Nile, dengue and other flaviviruses is the principal stimulus for the development of neutralizing antibodies and contains a fusion peptide responsible for inserting the virus into the host cell membrane. This glycoprotein lies flat on the surface of the virion and therefore only epitopes on the outer or lateral surface are important immunogens. Changes in antigen recognition after exposure of the virus to low pH have yielded clues to the fusion process. PMID- 12648939 TI - A matter of fitness. PMID- 12648940 TI - Evolution of virulence: adaptive or not? PMID- 12648942 TI - Enterobacterial adhesins and the case for studying SNPs in bacteria. AB - Single-nucleotide polymorphisms (SNPs) in structural genes can have a dramatic effect on the biology of whole organisms, from bacteria and viruses to mammals. Here, we underscore the importance of SNPs in bacterial genes that contribute to the ability of pathogens to cause disease. SNPs that confer an adaptive advantage for bacterial pathogens have been discovered in the genes encoding the FimH and Dr adhesins of Escherichia coli and, most recently, Salmonella enterica sv. Typhimurium FimH. PMID- 12648943 TI - Will the enigma of Francisella tularensis virulence soon be solved? AB - Francisella tularensis is one of the most infectious bacterial pathogens known and is the causative agent of the zoonotic disease tularemia. In spite of the importance of this pathogen little is known about its virulence mechanisms. However, it is clear that the bacterium is an intracellular pathogen, replicating mainly in macrophages, with replication in amoebae also having been reported. The genome sequence of a high virulence strain of F. tularensis is close to completion and when available, will stimulate further research into virulence mechanisms. PMID- 12648944 TI - Spying on HIV with SPR. AB - The application of surface plasmon resonance (SPR)-based optical biosensors has contributed extensively to our understanding of functional aspects of HIV. SPR biosensors allow the analysis of real-time interactions of any biomolecule, be it protein, nucleic acid, lipid, carbohydrate or small molecule, without the need for intrinsic or extrinsic probes. As such, the technology has been used to analyze molecular interactions associated with every aspect of the viral life cycle, from basic studies of binding events occurring during docking, replication, budding and maturation to applied research related to vaccine and inhibitory drug development. Along the way, SPR biosensors have provided a unique and detailed view into the inner workings of HIV. PMID- 12648946 TI - Exploiting host microtubule dynamics: a new aspect of bacterial invasion. AB - During infection, many pathogenic bacteria modulate the actin cytoskeleton of eukaryotic host cells to facilitate various infectious processes such as the attachment to or invasion of epithelial cells. Additionally, some pathogenic bacteria are capable of modulating the dynamics of host microtubule (MTs). Although the molecular basis for this is still poorly understood, a recent study of the Shigella VirA effector protein, which is delivered via a type III secretion system, suggests that MT destabilization plays an important role in Shigella infection. PMID- 12648945 TI - Helicobacter pylori virulence factors and the host immune response: implications for therapeutic vaccination. AB - Helicobacter pylori colonizes the human gastric mucosa and is associated with specific gastric disease. Virulence factors, such as urease, the vacuolating toxin (VacA), the cytotoxin-associated antigen CagA or blood-group-antigen binding adhesin (BabA), an adherence factor, might account for the development of different diseases. Vaccination trials exploiting the antigenic properties of some of these proteins have not been successful in preventing infection in humans. A more in-depth understanding of the immune response to H. pylori infection as well as additional information on suitable epitopes and adjuvants will be required before a successful vaccine can be developed. PMID- 12648947 TI - Changing anthropogenic influence on the Santa Monica Bay watershed. AB - Santa Monica Bay is an open coastal embayment located directly seaward of Los Angeles, California. The Bay provides vital economic value through its water dependent activities, such as swimming, diving, boating, and fishing. An increase from 100,000 residents in 1900 to 10 million in 2000 has imposed numerous environmental stressors on the Bay, including urbanization of the watershed. Pollutant discharges into the Bay increased throughout the early part of the century, but declined following passage of the Clean Water Act in 1972. Since that time, the predominant source of pollutant inputs has changed from point sources to non-point urban runoff. To assess how present-day and historical pollution interact to affect the environmental quality of Santa Monica Bay, three organizations collaborated on a multi-disciplinary study in 1997, towards which this volume is focused. This paper details the temporal patterns of anthropogenic influence on Santa Monica Bay to provide context for the papers that follow. PMID- 12648948 TI - Geomorphology, acoustic backscatter, and processes in Santa Monica Bay from multibeam mapping. AB - Santa Monica Bay was mapped in 1996 using a high-resolution multibeam system, providing the first substantial update of the submarine geomorphology since the initial compilation by Shepard and Emery [(1941) Geol. Soc. Amer. Spec. Paper 31]. The multibeam mapping generated not only high-resolution bathymetry, but also coregistered, calibrated acoustic backscatter at 95 kHz. The geomorphology has been subdivided into six provinces; shelf, marginal plateau, submarine canyon, basin slope, apron, and basin. The dimensions, gradients, and backscatter characteristics of each province is described and related to a combination of tectonics, climate, sea level, and sediment supply. Fluctuations of eustatic sea level have had a profound effect on the area; by periodically eroding the surface of Santa Monica plateau, extending the mouth of the Los Angeles River to various locations along the shelf break, and by connecting submarine canyons to rivers. A wetter glacial climate undoubtedly generated more sediment to the rivers that then transported the increased sediment load to the low-stand coastline and canyon heads. The trends of Santa Monica Canyon and several bathymetric highs suggest a complex tectonic stress field that has controlled the various segments. There is no geomorphic evidence to suggest Redondo Canyon is fault controlled. The San Pedro fault can be extended more than 30 km to the northwest by the alignment of a series of bathymetric highs and abrupt changes in direction of channel thalwegs. PMID- 12648950 TI - Spatial analysis of grain size in Santa Monica Bay. AB - Maps are useful scientific tools for presenting environmental information, but the statistical techniques necessary to prepare scientifically rigorous maps have primarily focused on terrestrial habitats. This study compares three popular techniques (triangulation, kriging, and co-kriging) to map sediment grain size in Santa Monica Bay, California. Two grain size data sets, one collected in 1994 (79 sites) and one collected in 1997 and 1998 (149 sites) were used for model development. A bathymetric data set collected in 1997 was used as a model covariate. A third grain size data set (40 sites) collected in 1996 from independent sites was used for model evaluation. Predictions were compared to validation data by average difference, prediction mean square error (PMSE), and a goodness-of-prediction measure, G. The average difference between prediction and truth was similar for all methods, but the PMSE for triangulation was more than twice that for kriging or co-kriging, which were similar. The G measure also shows triangulation to be a far worse predictor than kriging and co-kriging. Small-scale differences were observed between kriging and co-kriging at steep depth contours, where co-kriging predicted values commensurate with the expected depth-defined grain size. PMID- 12648949 TI - Characterizing benthic substrates of Santa Monica Bay with seafloor photography and multibeam sonar imagery. AB - Seafloor photography from three cruises is combined with multibeam sonar imagery to characterize benthic substrates and associated fauna of Santa Monica Bay, California. The multibeam EM1000 imagery was collected in 1996. Two sampling cruises (in 1998 and 1999) provided photographs at 142 sites throughout the Bay; a final cruise (in 2000) collected still photographs and continuous video along nine transects on the mainland shelf from Pt. Dume to the Palos Verdes peninsula. Muddy substrates (typically low backscatter) were the predominant habitat throughout the Santa Monica Bay, from the 20 m isobath to the adjacent Santa Monica basin floor (780 m). Bioturbation was pervasive as evidenced by abundant open burrows, mounds, and faunal tracks and trails. Sandy substrates (typically intermediate to high backscatter) were restricted to the innermost mainland shelf and a narrow outer shelf band north of Santa Monica Canyon. Cobble and gravel substrates (high backscatter) were restricted to the innermost shelf south of El Segundo and limited parts of the shelf edge. Rocky substrates (high backscatter) with interspersed patches of sand and gravel occurred on the high-relief marginal plateau and along parts of the shelf break offshore of Malibu. PMID- 12648951 TI - Clay-mineral suites, sources, and inferred dispersal routes: Southern California continental shelf. AB - Clay mineralogy is useful in determining the distribution, sources, and dispersal routes of fine-grained sediments. In addition, clay minerals, especially smectite, may control the degree to which contaminants are adsorbed by the sediment. We analyzed 250 shelf sediment samples, 24 river-suspended-sediment samples, and 12 river-bed samples for clay-mineral contents in the Southern California Borderland from Point Conception to the Mexico border. In addition, six samples were analyzed from the Palos Verdes Headland in order to characterize the clay minerals contributed to the offshore from that point source. The <2 microm-size fraction was isolated, Mg-saturated, and glycolated before analysis by X-ray diffraction. Semi-quantitative percentages of smectite, illite, and kaolinite plus chlorite were calculated using peak areas and standard weighting factors. Most fine-grained sediment is supplied to the shelf by rivers during major winter storms, especially during El Nino years. The largest sediment fluxes to the region are from the Santa Ynez and Santa Clara Rivers, which drain the Transverse Ranges. The mean clay-mineral suite for the entire shelf sediment data set (26% smectite, 50% illite, 24% kaolinite+chlorite) is closely comparable to that for the mean of all the rivers (31% smectite, 49% illite, 20% kaolinite+chlorite), indicating that the main source of shelf fine-grained sediments is the adjacent rivers. However, regional variations do exist and the shelf is divided into four provinces with characteristic clay-mineral suites. The means of the clay-mineral suites of the two southernmost provinces are within analytical error of the mineral suites of adjacent rivers. The next province to the north includes Santa Monica Bay and has a suite of clay minerals derived from mixing of fine-grained sediments from several sources, both from the north and south. The northernmost province clay-mineral suite matches moderately well that of the adjacent rivers, but does indicate some mixing from sources in adjacent provinces. PMID- 12648952 TI - Spatial scales and evolution of stormwater plumes in Santa Monica Bay. AB - Rainfall during winter storms produces extensive turbid, freshwater plumes in the coastal waters of the Southern California Bight. When the plumes result from urban runoff they contain toxic pollutants along with pathogenic bacteria and viruses, often resulting in closure of public beaches. We examined the spatial structure and evolution of stormwater plumes in Santa Monica Bay in 1996. The plumes resulted from freshwater discharge from the Ballona Creek and Malibu Creek watersheds which supply approximately 60% of the freshwater runoff to Santa Monica Bay. The spatial scales of the plumes were determined using shipboard measurements of water properties obtained from towyo transects and surface underway sampling. Salinity maps showed that the plumes typically extended 4-7 km offshore, consistent with scaling by the internal Rossby radius of deformation. Plumes extended along shore 10 km or more. Generally the plumes occupied the upper 10 m of the water column. The persistence time of a plume offshore of Ballona Creek was about three days based on a sequence of surveys in March 1996 following rainfall of about 21 mm. Limited comparison of plumes from Ballona Creek, which drains a developed watershed, and Malibu Creek, which drains a rural watershed, suggested that Malibu Creek required greater rainfall to produce an offshore plume. A stormwater plume offshore of Malibu Creek was observed on both sides of the creek mouth, possibly due to freshwater discharge from smaller surrounding watersheds or advection of freshwater discharges from the east and south. Plumes offshore of Ballona Creek mainly resulted from the creek itself and usually extended northward from the creek mouth, consistent with the wind forcing and the Coriolis acceleration. PMID- 12648953 TI - Observations of large-amplitude cross-shore internal bores near the shelf break, Santa Monica Bay, CA. AB - Two sets of moorings were deployed along a cross-shelf transect in central Santa Monica bay for four months in the winter of 1998-1999. Both sites had an array of instruments attached to tripods set on the seafloor to monitor currents over the entire water column, surface waves, near-bed temperature, water clarity and suspended sediment. A companion mooring had temperature sensors spaced approximately 10 m apart to measure temperature profiles between the surface and the seafloor. One array was deployed in 70 m of water at a site adjacent to the shelf break, just northwest of a major ocean outfall. The other was deployed on the mid shelf in 35 m of water approximately 6 km from the shelf break site. The subtidal currents in the region flowed parallel to the isobaths with fluctuating time scales around 10 days, a typical coastal-ocean pattern. However, during the falling phase of the barotropic spring tide, sets of large-amplitude, sheared cross-shore current pulses with a duration of 2-5 h were observed at the shelf break site. Currents in these pulses flowed exclusively offshore in a thin layer near the bed with amplitudes reaching 30-40 cm/s. Simultaneously, currents with amplitudes around 15-20 cm/s flowed exclusively onshore in the thicker layer between the offshore flow layer and the sea surface. The net offshore transport was about half the onshore transport. Near-surface isotherms were depressed 30-40 m. These pulses were likely internal bores generated by tidal currents. Bed stresses associated with these events exceeded 3 dynes/cm(2). These amplitudes are large enough to resuspend and transport not only fine-grained material, but also medium to coarse sands from the shelf toward the slope. Consequently, the seafloor over the shelf break was swept clear of fine sediments. The data suggest that the internal bores dissipate and are reduced in amplitude as they propagate across this relatively narrow shelf. There is evidence that they reach the 35 m site, but other coastal ocean processes obscure their distinctive characteristics. PMID- 12648954 TI - Magnitude and variability of Holocene sediment accumulation in Santa Monica Bay, California. AB - The spatial variability of Holocene (past 10,000 years) sediment accumulation in Santa Monica Bay (California) was examined to identify controls sediment trapping in a bathymetrically complex coastal embayment and to provide geologic context for the post-industrial sedimentary record and associated pollution gradients. Sediment chronologies based on downcore AMS 14C dates were used to quantify long term (millennia) accumulation rates in an effort to elucidate particle-transport pathways and sinks. Sediment accumulation rates for the full range of bayfloor environments (50-630 m water depths) range from 22 to 102 mg/cm(2)/year (15-88 mm/100 year), have an overall mean of 51+/-21 mg/cm(2)/year (1 sigma, n=11), and are comparable to rates reported for adjacent borderland basins. Maximal accumulation rates on the Malibu shelf and within a reentrant to Redondo canyon are interpreted to reflect (1) proximity to sediment sources and (2) localized oceanographic and topographic conditions conducive to sediment trapping and deposition. The 14C-derived accumulation rates are 2-10 times lower than rates determined through (210)Pb geochronology for the same sites in a related study, revealing that Holocene sediment accumulation has been non-steady-state. Santa Monica Bay is an important sink for suspended matter; averaged over the past several millennia a mass of sediment equivalent to 10-80% of the modern annual river supply is sequestered yearly. Net influx of suspended matter derived from the adjacent Palos Verdes shelf is evinced by a concentration gradient of p,p' DDE in bayfloor sediments, whereas the distribution of anthropogenic silver suggests transport from Santa Monica shelf to the southeastern boundary of the bay. The results of this study provide new insight to the long-term fates of particulate matter in Los Angeles coastal waters. PMID- 12648955 TI - Modern sedimentary processes in the Santa Monica, California continental margin: sediment accumulation, mixing and budget. AB - Sediment input to SMB appears to be associated with at least two point sources on the shelf, with Malibu Creek and the Hyperion sewage outfall being the most significant. Sediment contributions are sufficient to support apparent mass accumulation rates near these sources up to approximately 1.8 g/cm(2) year, which with distance decrease to approximately 0.5 g/cm(2) year near the shelf break (approximately 80-100 m water depth). Sequestering of material on the shelf and decreasing sediment supply to the slope is evident as rates decrease between 100 and 200 m water depths to less than 0.2 g/cm(2) year. Below 100-200 m water depth, rates are relatively slow throughout a broad region of the slope (0.07 0.14 g/cm(2) year). These slower rates are in general agreement with rates determined on the flanks of the California Borderland basins. Sediment texture fines from approximately 3.5 phi to approximately 7 phi with distance offshore. Texture does not exhibit significant changes from surficial values with depth in the seabed at any given site or between sites on the slope. This similarity in rates and downcore texture over such a broad extent suggests that hemiplegic sedimentation is the dominant mechanism of sediment delivery in water depths >200 m. Seabed distributions of radionuclides suggest that apparent accumulation rates in SMB may be twice the actual accumulation rates. A sediment budget documents that over the past century at least, SMB has served as a sink for 50-100% of the natural and anthropogenic inputs to the coastal ocean. PMID- 12648956 TI - Water quality impacts of stormwater discharges to Santa Monica Bay. AB - Urban stormwater runoff is a major source of contaminants to southern California's coastal waters, yet little is known about the fate and effects of these discharges. A 3-year multidisciplinary project was conducted to investigate the dispersion of stormwater plumes in Santa Monica Bay and the resultant impacts on the water column and benthos. This paper describes the toxicity component of the study. Sea urchin fertilization toxicity tests were conducted on stormwater from the two largest discharges into the bay: Ballona Creek, which drains a highly urbanized watershed, and Malibu Creek, which receives runoff from a largely undeveloped watershed. Every sample of Ballona Creek stormwater tested was toxic (usually >5 toxic units), while Malibu Creek stormwater had a lower frequency and magnitude of toxicity (usually <4 toxic units). Surface water samples collected within the Ballona Creek stormwater discharge plume were always toxic whenever the concentration of stormwater in the plume exceeded 10%. The toxic portion of the Ballona Creek stormwater plume extended more than 4 km offshore on one occasion. Toxicity identification studies indicated that zinc was the primary cause of toxicity in both Ballona Creek stormwater and the discharge plume. No acute sediment toxicity (10-day amphipod survival) was present in the study area, although interstitial water toxicity was present at some stations located near the mouth of Ballona Creek. Differences in watershed characteristics likely were responsible for the greater toxicity of the Ballona Creek stormwater discharge plume. The Ballona Creek watershed contained a greater degree of urbanization (83% versus 12% for Malibu Creek) and the presence of a network of concrete flood control channels resulted in a stormwater plume containing elevated concentrations of toxins that received less initial dilution (compared to Malibu Creek) in the nearshore environment. PMID- 12648957 TI - Impacts of stormwater discharges on the nearshore benthic environment of Santa Monica Bay. AB - Although large loads of potentially toxic constituents are discharged from coastal urban watersheds, very little is known about the fates and eventual impacts of these stormwater inputs once they enter the ocean. The goal of this study was to examine the effects of stormwater discharges on the benthic marine environment of Santa Monica Bay. Sediment samples were collected across a gradient of stormwater impact following significantly sized storm events offshore Ballona Creek (a predominantly developed watershed) and Malibu Creek (a predominantly undeveloped watershed). Sediments offshore Malibu Creek had a greater proportion of fine-grained sediments, organic carbon, and naturally occurring metals (i.e., aluminum and iron), whereas sediments offshore Ballona Creek had higher concentrations of anthropogenic metals (i.e., lead) and organic pollutants (i.e., total DDT, total PCB, total PAH). The accumulation of anthropogenic sediment contaminants offshore Ballona Creek was evident up to 2 km downcoast and 4 km upcoast from the creek mouth and sediment concentrations covaried with distance from the discharge. Although changes in sediment texture, organic content, and an increase in sediment contamination were observed, there was little or no alteration to the benthic communities offshore either Ballona or Malibu Creek. Both sites were characterized as having an abundance, species richness, biodiversity and benthic response index similar to shallow water areas distant from creek mouths throughout the Southern California Bight. There was not a preponderance of pollution tolerant, nor a lack or pollution sensitive, species offshore either creek mouth. PMID- 12648958 TI - Retrospective evaluation of shoreline water quality along Santa Monica Bay beaches. AB - Santa Monica Bay (SMB) beaches are the most heavily used in the U.S.A., despite an increased number of water quality postings over the last several years. To assess whether water quality problems are concentrated at a small number of chronically affected sites or whether the problems are widely distributed, we compiled 5 years of monitoring data collected at 59 sites, 22 of which are sampled daily. Other locally available rainfall and sewage spill monitoring information data were added to this data set to assess whether sewage spills, dry weather runoff, or wet-weather runoff contribute the most to exceedences of water quality thresholds. Approximately 13% of the shoreline mile-days along monitored beaches in SMB exceeded the State of California's beach water quality standards during the 5-year study period. Most of the water quality exceedences occurred near urban runoff drains even though areas affected by drains represent only a small portion of the total shoreline. Although storms are relatively infrequent in southern California, the extent of water quality exceedences resulting from storm water runoff was similar to the extent of water quality exceedences found during dry weather. Sewage spills, while potentially more serious because they lead to beach closures rather than to the more limited posting of warning signs, represented less than 0.1% of the shoreline mile-days that exceeded water quality thresholds. During dry weather conditions, most of the water quality problems occurred near five of the largest drains and at two beach areas that have unique physical characteristics, which limited mixing, dispersion, and dilution. During wet weather conditions, water quality problems were more widespread. PMID- 12648959 TI - Temporal and spatial distributions of contaminants in sediments of Santa Monica Bay, California. AB - Contaminant inputs from wastewater discharge, a major source of contamination to Santa Monica Bay (SMB), have declined drastically during the last three decades as a result of improved treatment processes and better source control. To assess the concomitant temporal changes in the SMB sediments, a study was initiated in June 1997, in which 25 box cores were collected using a stratified random sampling design. Five sediment strata corresponding to the time intervals of 1890 1920, 1932-1963, 1965-1979, 1979-1989, and 1989-1997 were identified using (210)Pb dating techniques. Samples from each stratum were analyzed for metals, 1,1,1-Trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) and its metabolites (DDTs), polychlorinated biphenyls (PCBs), and total organic carbon (TOC). Samples from the 1965-1979, 1979-1989, and 1989-1997 strata were also analyzed for polycyclic aromatic hydrocarbons (PAHs) and linear alkylbenzenes (LABs). Sediment metal concentrations increased from 1890-1979 and were similar during the time intervals of 1965-1979, 1979-1989, and 1989-1997, although the mass emissions of trace metals from sewage inputs declined substantially during the same time period. Trace organic contamination in SMB was generally highest in sediments corresponding to deposition during the years of 1965-1979 or 1979-1989 and showed a decline in concentration in the 1989-1997 stratum. Temporal trends of contamination were greatest in sediments collected from areas near the Hyperion Treatment Plant (HTP) outfall system and on the slope of Redondo Canyon. The highest contaminant concentrations were present in sediments near the HTP 7-mile outfall in the 1965-1979 stratum. Elevated trace metal and organic concentrations were still present in the 1989-1997 stratum of most stations, suggesting that sediment contaminants have moved vertically in the sediment column since sludge discharges from the 7-mile outfall (a dominant source of contamination to the bay) ceased in 1987. The widespread distributions of DDTs and PCBs in SMB and highly confined distribution of LABs around the HTP outfall system were indicative of a dispersal mechanism remobilizing historically deposited contaminants to areas relatively remote from the point of discharge. PMID- 12648961 TI - Response of benthic foraminifers to sewage discharge and remediation in Santa Monica Bay, California. AB - Examination of a time series of foraminiferal assemblage distributions on the continental shelf and slope of Santa Monica Bay from 1955 to 1997-1998 suggests that the benthic microfauna have been greatly affected by the quality and character of the municipal sludge and wastewater discharged into the bay over the last half-century by the Hyperion Treatment Plant serving the greater Los Angeles area. Five species dominate both the living and dead foraminiferal assemblages of the 1997-1998 surface samples, including Eggerella advena, Trochammina pacifica, Bulimina denudata, Buliminella elegantissima, and Epistominella bradyana. Temporal patterns of relative species abundances for both living and dead assemblages, as well as toxicity tests measuring amphipod survival and sea urchin fertilization success, show improvement since the sewage treatment program was enhanced in 1986. None of these trends are evident 10 years earlier, coincident with the onset of a Pacific Decadal Oscillation warming trend. This fact suggests that remediation, and not climate change, is responsible for the faunal changes observed. Even with remediation, however, all foraminiferal faunal trends have not returned to early-outfall levels. The organic-waste indicating species T. pacifica shows a slow decline in abundance as sewage treatment and sludge disposal activities have improved, whereas a dramatic increase in the abundance of the pioneer colonizer of impacted regions, E. advena, has occurred, often with a reciprocal response by B. denudata. Also evident is a dramatic shift in the abundance of the once-dominant species Nonionella basispinata and Nonionella stella, which were unable to recolonize Santa Monica Bay since the two major outfalls (5- and 7-mile) began discharging. Temporal variations in species abundances, as well as range expansions, contractions, and the inability to recolonize areas previously, or presently, impacted, suggests that foraminifers are a useful tool in defining areas affected by waste discharge. PMID- 12648962 TI - "Pro-life" policy threatens US HIV/AIDS initiative. PMID- 12648960 TI - Toxicity assessment of sediment cores from Santa Monica Bay, California. AB - During the summer of 1997, sediment core samples were taken at 25 stations in Santa Monica Bay. Toxicity testing was performed on 4-cm sections of the entire length of each core using purple sea urchin fertilization and amphipod survival tests. The sea urchin test identified sections as being toxic at six stations, all located near current or former Hyperion Treatment Plant (HTP) wastewater outfall locations. The amphipod test identified sections from 17 stations as having toxic sediments. The stations having toxic sediments were scattered throughout the bay and toxicity was identified at numerous core depths. Spatial and temporal patterns indicated that toxicity was most strongly associated with the historical disposal of municipal wastewater sludge. Many of the sections toxic to the amphipods did not have chemical levels expected to cause toxicity and were in locations where a source of toxicity was not apparent. PMID- 12648963 TI - Dementia comes of age in the developing world. PMID- 12648964 TI - Neuromyelitis optica: what it is and what it might be. PMID- 12648965 TI - Human metapneumovirus in the community. PMID- 12648966 TI - Genomics--a global public good? PMID- 12648967 TI - Postnatal home visits in teenage mothers: a randomised controlled trial. AB - BACKGROUND: Teenage pregnancies are associated with negative socioeconomic effects. Our aim was to ascertain whether a postnatal home-visiting service for teenage mothers younger than age 18 years could reduce the frequency of adverse neonatal outcomes and improve knowledge of contraception, breastfeeding, and infant vaccination schedules in this parent group. METHODS: We enrolled 139 adolescents, attending a teenage pregnancy clinic, in a randomised controlled trial. After completing an antenatal questionnaire designed to assess their knowledge of contraception, infant vaccination, and breastfeeding, we assigned participants to either receive five structured postnatal home visits by nurse midwives (n=65) or not (n=71). Assessment interviews were done 6 months postpartum. Our primary endpoint was unadjusted difference in knowledge between groups, and incidence of predefined adverse neonatal outcomes. Analysis was by intention to treat. FINDINGS: Three women withdrew before randomisation because of late fetal loss, 11 mothers withdrew because of adverse neonatal outcomes (adverse neonatal outcome was a primary endpoint, but resulted in withdrawal from the study for knowledge outcomes), and one left voluntarily. Follow-up data were, therefore, available for 124 teenagers. Postnatal home visits were associated with a reduction in adverse neonatal outcomes (intervention: 2; control: 9; relative risk 0.24, 95% CI 0.05-1.08), and a significant increase in contraception knowledge (mean difference 0.92, 95% CI 0.32-1.52). However, there was no significant increase in knowledge with respect to breastfeeding or infant vaccination schedules associated with the home visits. INTERPRETATION: Postnatal home-visiting services by nurse-midwives reduce adverse neonatal events and improve contraception outcomes, but do not affect breastfeeding or infant vaccination knowledge or compliance. PMID- 12648968 TI - Thrombophilic disorders and fetal loss: a meta-analysis. AB - BACKGROUND: Our aim was to assess the strength of the controversial association between thrombophilia and fetal loss, and to examine whether it varies according to the timing or definition of fetal loss. METHODS: We searched Medline and Current Contents for articles published between 1975 and 2002 and their references with terms denoting recurrent fetal and non-recurrent fetal loss combined with various thrombophilic disorders. We included in our meta-analysis case-control, cohort, and cross-sectional studies published in English, the methodological quality of which was rated as moderate or strong. Pooled odds ratios (OR) with 95% CI were generated by random effects models with Cochrane Review Manager software. FINDINGS: We included 31 studies. Factor V Leiden was associated with early (OR 2.01, 95% CI 1.13-3.58) and late (7.83, 2.83-21.67) recurrent fetal loss, and late non-recurrent fetal loss (3.26, 1.82-5.83). Exclusion of women with other pathologies that could explain fetal loss strengthened the association between Factor V Leiden and recurrent fetal loss. Activated protein C resistance was associated with early recurrent fetal loss (3.48, 1.58-7.69), and prothrombin G20210A mutation with early recurrent (2.56, 1.04-.29) and late non-recurrent (2.30, 1.09-4.87) fetal loss. Protein S deficiency was associated with recurrent fetal loss (14.72, 0.99-218.01) and late non-recurrent fetal loss (7.39, 1.28-42.63). Methylenetetrahydrofolate mutation, protein C, and antithrombin deficiencies were not significantly associated with fetal loss. INTERPRETATION: The magnitude of the association between thrombophilia and fetal loss varies, according to type of fetal loss and type of thrombophilia. PMID- 12648969 TI - Dementia diagnosis in developing countries: a cross-cultural validation study. AB - BACKGROUND: Research into dementia is needed in developing countries. Assessment of variations in disease frequency between regions might enhance our understanding of the disease, but methodological difficulties need to be addressed. We aimed to develop and test a culturally and educationally unbiased diagnostic instrument for dementia. METHODS: In a multicentre study, the 10/66 Dementia Research Group interviewed 2885 people aged 60 years and older in 25 centres, most in Universities, in India, China and southeast Asia, Latin America and the Caribbean, and Africa. 729 had dementia and three groups were free of dementia: 702 had depression, 694 had high education (as defined by each centre), and 760 had low education (as defined by each centre). Local clinicians diagnosed dementia and depression. An interviewer, masked to dementia diagnosis, administered the geriatric mental state, the community screening instrument for dementia, and the modified Consortium to Establish a Registry of Alzheimer's Disease (CERAD) ten-word list-learning task. FINDINGS: Each measure independently predicted a diagnosis of dementia. In an analysis of half the sample, an algorithm derived from all three measures gave better results than any individual measure. Applied to the other half of the sample, this algorithm identified 94% of dementia cases with false-positive rates of 15%, 3%, and 6% in the depression, high education, and low education groups, respectively. INTERPRETATION: Our algorithm is a sound basis for culturally and educationally sensitive dementia diagnosis in clinical and population-based research, supported by translations of its constituent measures into most languages used in the developing world. PMID- 12648970 TI - Informed consent during the clinical emergency of acute myocardial infarction (HERO-2 consent substudy): a prospective observational study. AB - BACKGROUND: Anxiety, fear, pain, and treatment with morphine might compromise the ability of patients to comprehend information about, and give informed consent for, participation in clinical trials. We aimed to assess whether patients with acute myocardial infarction could understand written and verbal information and whether they were competent to give autonomous informed consent to participate in a clinical trial. METHODS: We prospectively studied 399 patients with acute myocardial infarction in 16 hospitals in New Zealand and Australia who were eligible for participation in the Hirulog and Early Reperfusion or Occlusion (HERO)-2 trial. We assessed readability of patient information sheets, patients' educational status, their views of the consent process, comprehension of verbal and written information, and competence to give consent. FINDINGS: The patient information sheet needed a year 13 (age 18) educational level for comprehension, although only 75 of 345 patients (22%) had been educated beyond secondary school. Only 63 of 346 (18%) read the patient information sheet before giving or refusing consent to participate. Patients who gave consent were more likely to report good or partial comprehension of the information provided than were those who refused consent (272 [89%] vs 14 [70%], respectively; p=0.009). In an assessment of competence to make an autonomous decision, 75 of 145 (52%) were ranked at the lowest grade and 26 (18%) were not competent to consent. INTERPRETATION: Although the consent process for HERO-2 met regulatory requirements for clinical trials, it was inappropriate for the needs of most patients. The patients' comprehension of the information provided and their competence to autonomously give consent was less than optimum. PMID- 12648971 TI - Septic metastatic endophthalmitis. PMID- 12648972 TI - Oligonucleotide microarray for prediction of early intrahepatic recurrence of hepatocellular carcinoma after curative resection. AB - BACKGROUND: Hepatocellular carcinoma has a poor prognosis because of the high intrahepatic recurrence rate. There are technological limitations to traditional methods such as TNM staging for accurate prediction of recurrence, suggesting that new techniques are needed. METHODS: We investigated mRNA expression profiles in tissue specimens from a training set, comprising 33 patients with hepatocellular carcinoma, with high-density oligonucleotide microarrays representing about 6000 genes. We used this training set in a supervised learning manner to construct a predictive system, consisting of 12 genes, with the Fisher linear classifier. We then compared the predictive performance of our system with that of a predictive system with a support vector machine (SVM-based system) on a blinded set of samples from 27 newly enrolled patients. FINDINGS: Early intrahepatic recurrence within 1 year after curative surgery occurred in 12 (36%) and eight (30%) patients in the training and blinded sets, respectively. Our system correctly predicted early intrahepatic recurrence or non-recurrence in 25 (93%) of 27 samples in the blinded set and had a positive predictive value of 88% and a negative predictive value of 95%. By contrast, the SVM-based system predicted early intrahepatic recurrence or non-recurrence correctly in only 16 (60%) individuals in the blinded set, and the result yielded a positive predictive value of only 38% and a negative predictive value of 79%. INTERPRETATION: Our system predicted early intrahepatic recurrence or non recurrence for patients with hepatocellular carcinoma much more accurately than the SVM-based system, suggesting that our system could serve as a new method for characterising the metastatic potential of hepatocellular carcinoma. PMID- 12648973 TI - Acute ophthalmoplegia. PMID- 12648974 TI - Bronchoscopic volume reduction with valve implants in patients with severe emphysema. AB - Eight patients with severe emphysema entered a pilot study of unilateral volume reduction by endobronchial valve insertion. Five patients had emphysema judged too severe for volume reduction surgery and three refused the operation. After valve insertions, the median forced expiratory volume in 1 s (FEV1) increased from 0.79 L (range 0.61-1.07) to 1.06 L (0.75-1.22) (difference 34%, p=0.028) and the median diffusing capacity (TL(CO)) increased from 3.05 mL/min/mm Hg (2.35 4.71) to 3.92 mL/min/mm Hg (2.89-5.40) (difference 29%, p=0.017). CT scans showed a substantial reduction in regional volume in four of the eight patients. Two patients developed a transient pneumothorax (one requiring drainage) but we recorded no other important adverse effects during follow-up. Lung-volume reduction can be achieved with unilateral bronchoscopically placed valve implants in patients with severe emphysema with acceptable short-term safety and worthwhile functional benefits. PMID- 12648976 TI - Experts gather to discuss water crisis that the world is ignoring. PMID- 12648975 TI - Chronic synovitis and HLA B27 in patients with severe haemophilia. AB - Chronic synovitis affects about 10% of patients with severe haemophilia in India. This disease has some features in common with ankylosing spondylitis, which has been linked to HLA B27. We therefore aimed to test whether there is an association between HLA B27 and chronic synovitis. We studied 473 patients with severe haemophilia (33 of whom had chronic synovitis), and 1175 healthy controls using a standard serological technique and the reverse line strip assay. 64% (21 of 33) of patients with haemophilia and chronic synovitis were positive for HLA B27, compared with 5% (23 of 440) of those with severe haemophilia, but not chronic synovitis (odds ratio 31.6 [95% CI 9.28-39.38], p<0.0001), and 9% (100 of 1175) of healthy controls (18.81 [9.6-27.7], p<0.0001). We conclude that there is a strong association between HLA B27 and chronic synovitis in Indian patients with severe haemophilia and screening in this population could allow treatment and prevention of the complication. PMID- 12648977 TI - Scientists grieve loss of astronauts and science. PMID- 12648978 TI - Uninsured people in USA put a strain on health system. PMID- 12648980 TI - Bronchiolitis drugs lack convincing evidence of efficacy. PMID- 12648983 TI - Uganda and the Global Fund sign grant agreement. PMID- 12648984 TI - Hodgkin's lymphoma. AB - Hodgkin's lymphoma was first described in 1832, but the nature of the pathognomic Reed-Sternberg cell, on which diagnosis of the disease is based, has only been elucidated in the past few years. Radiotherapy has been used to treat localised disease since the 1940s, and in the 1960s, effective combination chemotherapy regimens were introduced for anatomically advanced disease. The past three decades have witnessed continued improvement in outcome to such an extent that Hodgkin's lymphoma is now one of the most curable of all non-cutaneous malignancies. With improved survival and extended follow-up, relevance of treatment-induced late effects has become apparent, and modern therapeutic strategies must fully account for these effects. We review the pathology of Hodgkin's lymphoma, and its clinical presentation, investigation, present management, and natural history, including late effects of treatment. PMID- 12648985 TI - Prevention of variceal rebleeding. AB - CONTEXT: Variceal bleeding is the most frequent severe complication of portal hypertension and a leading cause of death and liver transplantation in patients with cirrhosis. Patients surviving a variceal bleed are at high risk of rebleeding (over 60% at 1 year). Portacaval shunts and transjugular intrahepatic portasystemic shunts (TIPS) are effective for prevention of rebleeding but carry a high risk of hepatic encephalopathy. Endoscopic techniques include band ligation (EBL) and injection sclerotherapy (EIS). Drug approaches are based on non-selective beta blocker with or without isosorbide-5-mononitrate (ISMN). STARTING POINT: David Patch and colleagues (Gastroenterology 2002; 123: 1013-19) randomised 102 patients surviving a variceal bleeding to EBL or drug therapy with propranolol with the addition of ISMN if target reductions in portal pressure (evaluated by the hepatic venous pressure-gradient [HVPG]) were not achieved at 3 months. Overall, results of drug therapy were similar to those of EBL (44% vs 54% rebleeding at 1 year). There were no differences in survival or non-bleeding complications. Christophe Bureau and colleagues (Hepatology 2002; 36: 1361-66) treated 34 patients with cirrhosis and portal hypertension with propranolol and measured HVPG after a median of 4 days. Target HVPG reductions were achieved in 13 "responders". ISMN was added in the 21 "non-responders" and HVPG measured again: seven more patients achieved target HPVG reduction. Re-bleeding rates were lower in responders than in non-responders (10% vs 64%). Both studies suggest that drug therapy can be improved by adding ISMN to b blockers in those patients with an insufficient decrease in HVPG. WHERE NEXT? Long-term drug therapy is emerging as effective treatment for the prevention of variceal rebleeding. The role of HVPG monitoring as a guide to identifying patients requiring further treatment needs to be further evaluated. Trials to determine the best treatment for patients who do not respond to drug therapies are also required. PMID- 12648986 TI - Pathological and molecular aspects of prostate cancer. AB - This review focuses on new findings and controversial issues in the the pathology and molecular biology of adenocarcinoma of the prostate. Since management of high grade prostatic intraepithelial neoplasia on needle biopsy--the most common precursor lesion to prostate cancer--is the crucial issue with this lesion, we discuss the risk of cancer subsequent to this histological diagnosis and the issue of whether such neoplasia should be regarded as carcinoma-in-situ. We also look at prostate cancer itself, starting with its diagnosis, reporting on needle biopsy, and reviewing how the most frequently used grading system, the Gleason grading system, affects treatment. The molecular basis of prostate cancer includes inheritable and somatic genetic changes (tumour suppressor genes, loss of heterozygosity, gene targets and regions of chromosomal gain, CpG island promoter methylation, invasion and metastasis suppressor genes, telomere shortening, and genetic instability). Changed gene expression (eg, proliferation related genes, changes in the androgen receptor, apoptosis and stress-response genes) have potential as biomarkers and therapeutic targets in prostate cancer. PMID- 12648987 TI - Open letter to Jong Wook Lee, Director-General elect of WHO. PMID- 12648988 TI - Primary angioplasty or thrombolysis for acute myocardial infarction? PMID- 12648989 TI - Primary angioplasty or thrombolysis for acute myocardial infarction? PMID- 12648990 TI - Primary angioplasty or thrombolysis for acute myocardial infarction? PMID- 12648991 TI - Primary angioplasty or thrombolysis for acute myocardial infarction? PMID- 12648993 TI - Dipyrone and agranulocytosis: what is the risk? PMID- 12648994 TI - Kidney transplantation with rabbit antithymocyte globulin and sirolimus monotherapy. PMID- 12648996 TI - Anticipation of epileptic seizures from standard EEG recordings. PMID- 12648998 TI - Gene expression profiling in biliary atresia. PMID- 12648999 TI - Anticipation of epileptic seizures from standard EEG recordings. PMID- 12649002 TI - Clinical research in Europe. PMID- 12649003 TI - Equipment for Ethiopian hospitals. PMID- 12649005 TI - Needs assessment for CME. PMID- 12649008 TI - Women on the treadmill. PMID- 12649010 TI - Kinematics of the human pelvis following open book injury. AB - The objective of this study is to determine the three dimensional kinematics of the human pelvis including both sacroiliac joints following a simulated open book injury induced on cadavers by applying anterior-posterior compressive loads to the pelvis. An electromagnetic digitizing and motion tracking system was utilized to measure the morphology of the pelvis and the relative movements of its bones during this simulated open book fracture. The screw displacement axis method was used to describe the relative motion between the sacrum and each hipbone. Morphologically, it was found that the articular surfaces forming the sacroiliac joints could be approximated with planar surfaces directed from proximal and lateral to distal and medial and from posteromedial to anterolateral. The kinematic data obtained from this study indicate that there is a direct correlation between the opening of the symphysis pubis and the opening of the sacroiliac joint (SIJ) during open book injury. This suggests that the extent of injury of the SIJ maybe estimated from the degree of opening of the symphysis pubis as demonstrated on anteroposterior (A-P) x-rays. The results obtained from this study also indicate that the motion of the hipbone with respect to the sacrum on the side of the sacroiliac joint opening is almost a pure rotation, which translates clinically on the A-P x-rays as pure opening of the SIJ without vertical displacement. The average axis of rotation was found to be almost parallel to the SIJ planar articular surface. Furthermore, the pubic bone on the side of SIJ opening was found to displace inferiorly and posteriorly. One can thus conclude that in open book pelvic injuries, the pubic bone on the side of injury displaces inferiorly on the outlet projection x-rays with no vertical displacement of the SIJ. This is important since the initial assessment of the open book injury in the emergency room includes outlet projection x-rays. From this study, the relative vertical positions of the pubic bones on these x-rays can help the surgeon in differentiating open book fracture injury from other pelvic injuries. PMID- 12649011 TI - Biomechanical investigation of pedicle screw-vertebrae complex: a finite element approach using bonded and contact interface conditions. AB - This study used finite element simulation to investigate the load transfer mechanisms within the screw/vertebra complex under different interface conditions, and under varying screw lengths. Both bonded and contact conditions were employed to demonstrate the interface between the screw and vertebra. Loadings were applied at the superior surface of the vertebra and screw unthreaded end, respectively, to represent two modes of flexion loads. The results indicated that the screw within the vertebra underwent a series of discontinuities of loading, identified by the localized high contact pressures, thus creating localized bending moments. The peak stress of screw was located at the junction of the screw's hub and thread, which is consistent with the location of screw failure observed in a clinical setting and the values of peak stress in the screw were proportional to the amount of moments generated by the two loading modes. The interface condition plays an important role in transferring the force within the screw/vertebra complex. A contact interface condition induces significantly higher stress in the screw than the bonded condition. Therefore providing a binding surface (with HA, or porosity coating on the screw surface) between the screw and the vertebra might be the most effective way to prevent screw failure. The influences of screw length on the peak stress in the screw become negligible when the screw is of sufficient length to extend fully into the vertebral body. PMID- 12649012 TI - How to determine the permeability for cement infiltration of osteoporotic cancellous bone. AB - Cement augmentation is an emerging surgical procedure in which bone cement is used to infiltrate and reinforce osteoporotic vertebrae. Although this infiltration procedure has been widely applied, it is performed empirically and little is known about the flow characteristics of cement during the injection process. We present a theoretical and experimental approach to investigate the intertrabecular bone permeability during the infiltration procedure. The cement permeability was considered to be dependent on time, bone porosity, and cement viscosity in our analysis. In order to determine the time-dependent permeability, ten cancellous bone cores were harvested from osteoporotic vertebrae, infiltrated with acrylic cement at a constant flow rate, and the pressure drop across the cores during the infiltration was measured. The viscosity dependence of the permeability was determined based on published experimental data. The theoretical model for the permeability as a function of bone porosity and time was then fit to the testing data. Our findings suggest that the intertrabecular bone permeability depends strongly on time. For instance, the initial permeability (60.89 mm(4)/N(*)s) reduced to approximately 63% of its original value within 18 seconds. This study is the first to analyze cement flow through osteoporotic bone. The theoretical and experimental models provided in this paper are generic. Thus, they can be used to systematically study and optimize the infiltration process for clinical practice. PMID- 12649013 TI - A method for in-vivo analysis for regional arterial wall material property alterations with atherosclerosis: preliminary results. AB - Atherosclerosis is a diffuse arterial disease developing over many years and resulting in a complicated three-dimensional arterial morphology. The arterial wall material properties have been demonstrated to show regional alterations with atheroma development and growth. We present a mechanical analysis of diseased arterial segments reconstructed from intravascular ultrasound images in order to quantitatively identify regional alterations in the elastic constants with atherosclerotic lesions. We employ a finite element and a displacement sensitivity analysis to divide the arterial segment into regions with different material properties and use an optimization algorithm to identify the elastic constants in these regions. The results with regional variations identified with this method correlated qualitatively with the extent and location of atherosclerotic lesions identified by visual inspection of the affected arteries. The optimized elastic modulus in regions affected by early atherosclerotic lesions ranged from 90.9 to 93.0 kPa where as the corresponding magnitudes in normal arterial segments ranged from 97.9 to 101.0 kPa. This method can be potentially employed to identify the extent and location of atherosclerotic lesions in a systematic analysis and may potentially be used for the early detection of lesion growth. PMID- 12649014 TI - Flow changes in the aorta associated with the deployment of a AAA stent graft. AB - The purpose of this study was to investigate the hemodynamic implications of a proximal shift in the aortic bifurcation that results from abdominal aortic aneurysm (AAA) stent graft deployment. A flow model was constructed in which an anatomically accurate model of the aorta was subjected to physiologic pulsatile flow. The model included the celiac, superior mesenteric, left and right renal arteries. The aortic bifurcation, leading to the right and left iliac arteries was included, as well as the lumbar curvature. Flow simulations were performed under resting and mild exercise conditions with and without a Cordis AAA stent graft deployed. Flow patterns were visualized with dye injection and recorded onto video. The flow rates through the iliac and renal arteries were continuously monitored using ultrasonic flowmeters. Flow visualization revealed that flow disturbances at the level of the renal arteries were slightly increased with the deployment of the stent graft. The orientation of the endolegs within the aorta had no perceptible effect on these disturbances. Under mild exercise conditions, very little flow disturbance was observed. In conclusion, there are slight changes in flow disturbance near the renal arteries due to stent graft deployment, but these changes would not be expected to have significant clinical implications. PMID- 12649015 TI - Lung function interpolation by means of neural-network-supported analysis of respiration sounds. AB - Respiration sounds of individual asthmatic patients were analysed in the scope of the development of a method for computerised recognition of the degree of airways obstruction. Respiration sounds were recorded during laboratory sessions of allergen provoked airways obstruction, during several stages of advancing obstruction. The technique of artificial neural networks was applied for relating sound spectra and simultaneously measured lung function values (spirometry parameter FEV(1)). The ability of feedforward neural networks was tested to interpolate obstruction levels of FEV(1)-classes of which no members were included in the set used for training a network. In this way, a situation was simulated of an existing network recognising a new asthmatic attack under the same physiological conditions. It appeared to be possible to interpolate FEV(1) values, and it is concluded that a deterministic relationship exists between sound spectra and lung function parameter FEV(1). Variance optimisation appeared to be important in optimising the neural network configuration. PMID- 12649016 TI - Arm EMG during abduction and adduction: hysteresis cycle. AB - The electromyographic study of the muscles involved in the complex movements of the shoulder, is usually one way to quantifying the static and dynamic joint's behavior. In particular, the deltoid medium EMG produced a phenomenon similar to a hysteresis cycle when its amplitude was plotted as a function of the lateral angular position during a static, step by step, sequential abduction-adduction of the arm. Such a cycle was consistently repeated in 16 subjects (12 males and 4 females). The paired Student t-test, after comparing the mean EMG values of the rectified wave for the same arm opening angle between abduction and adduction, produced a highly significant difference (alpha<0.001) in all subjects. In all likelihood, it manifests the participation of muscles other than the deltoid medium in the overall movement (as for example, the anterior and posterior deltoids), that is, they are collaborating muscles that are different in the opening or lifting of the arm from those involved in its closure or lowering. Thus, it is concluded that a quantifiable and significant deltoid medium EMG difference has been demonstrated when the muscle is either ready to abduction or ready to adduction. The effect is fully reproducible between 0 and 90 degrees of an arm in static position within the scapular plane. PMID- 12649018 TI - Dual channel light pulse exposure timer. AB - An instrument has been developed for routine measurement of pulse durations of lasers and pulsed light sources. A four digit light emitting diode (LED) count display is used with a range of selectable clock rates in five decades from 1 MHz to 100 Hz with a longest time duration corresponding to 99.99 s. The performance of the system was verified using a Schmidt trigger test circuit to drive an LED where the time of the drive pulse was compared to that indicated by the timer unit and where also silicon photodiodes of different detector areas were used to verify device measurement accuracy.A buffered output channel allows inspection by an oscilloscope to verify output pulse characteristics. This can be especially relevant to verify performance characteristics of a given laser/pulsed light system and to establish definitive assessments for eye safety based on relevant standards. PMID- 12649017 TI - Prediction of response to incision using the mutual information of electroencephalograms during anaesthesia. AB - This paper presents a new approach to predict response during isoflurane anaesthesia by using mutual information (MI) time series of electroencephalograms (EEGs) and their complexity analysis. The MI between four lead electrodes was first computed using the EEG time series. The Lempel-Ziv complexity measures, C(n)s, were extracted from the MI time series. Prediction was made by means of artificial neural network (ANN). From 98 consenting patient experiments, 98 distinct EEG recordings were collected prior to incision during isoflurane anaesthesia of different levels. During and after skin incision, each patient was observed carefully for 2 min to detect subsequent responses (purposeful movement, changes in hemodynamic parameters and respiratory pattern) and then the EEG was labelled as 0.0 for responder or as 1.0 for non-responder. Training and testing the ANN used the 'drop-one-patient' method. The prediction was tested by monitoring the response to incision and the result given by the ANN. The system was able to correctly classify purposeful response in average accuracy of 91.84% of the cases. The results showed that the method has a better performance than other methods, such as spectral edge frequency, median frequency, and bispectral analysis. This method is computationally fast and acceptable real-time clinical performance was obtained. PMID- 12649019 TI - An experimental two degrees-of-freedom actuated external fixator for in vivo investigation of fracture healing. AB - A novel two degrees-of-freedom external fixator that can accurately control interfragmentary mechanical conditions over an ovine tibial osteotomy is presented. The device can apply either axial or torsional motion, to investigate mechano-transduction mechanisms in fracture healing. The device was bench-tested over a range of gap stiffnesses, for its ability to apply pure axial or torsional motions, control interfragmentary strain magnitudes and to measure stiffness over the gap. The experiments reveal very little cross-coupling between axial and torsional displacements, although some transverse displacement occurs for both degrees-of-freedom. The mean interfragmentary strain can also be accurately controlled, but with some variation over the gap (due to coupled transverse displacement error). Interfragmentary stiffness measurements are accurate up to gap stiffnesses corresponding to bony bridging. This characterisation study has shown that the system is adequate to control and measure the mechanical conditions in a fracture healing experiment. PMID- 12649020 TI - Development and evaluation of an automated stainer for acid-fast bacilli. AB - The current strategy for the control of tuberculosis (TB) relies on early diagnosis, and smear microscopy is an essential component of the laboratory diagnosis of TB in most countries with a high prevalence of the disease. However, even simple smear microscopy examination is far from satisfactory because staining results can vary among individual technicians. In an effort to minimize variations in manual staining procedures, we developed an automated stainer for AFB and evaluated its usefulness in comparison with manual staining. The key feature of our automated stainer is a heating apparatus required for fixation and carbol-fuchsin staining. After smear slides are placed into the machine, the entire staining process is fully automated, from fixation to final washing and drying. With the automated methods, five slides can be fixed and stained in 21 min at consistent high quality. Using sputum samples from 91 TB patients, the staining results of the automated stainer were compared blindly with those of manual staining. The concordance rate between the two methods was 94.5%. In addition, there was no significant difference in the rate of detection of AFB in the sputum samples. Although further optimization of the auto staining procedures is required, the results indicate that the automated AFB stainer developed in this study looks promising for use in clinical mycobacteriology laboratory in order to minimize personal variation during AFB staining. PMID- 12649021 TI - Meniscus preservation; rationale, repair techniques and results. AB - It is inescapable that loss of part or all of the structure of the meniscus of the knee alters the biomechanics of knee function leading to a gradual deterioration of the chondral surface, which ultimately will result in osteoarthritis. The goal of the knee surgeon should therefore be to preserve as much as possible the function of the meniscus. Classic teaching is that many meniscal tears, which are peripheral and longtitudinal, will heal if sutured and protected during a slow healing process. However it is clear that other tear configurations can heal if given the opportunity by adequate preparation of the meniscus rim, careful suturing and delayed rehabilitation. Thus some radial tears, horn avulsions, double longtitudinal tears and retears, previously thought to be irreparable, can be sutured with good results. Over 12 years, 288 meniscus repairs were undertaken in 265 patients ranging in age from 12 to 70 years. Most were repaired at the time of anterior cruciate ligament reconstruction, although 55 were isolated repairs. While the average time from injury to surgery was 11.7 months (1 - 72), one third of patient's already had chondral damage of at least Outerbridge Grade 2. The purpose of this paper is to report on the more difficult meniscus repair techniques and as such formal follow up was not undertaken to determine the final outcome of these patients. Never the less it is estimated from known data that there was a retear rate in the order of less than 10%. There have been three complications, 2 infections and one temporary neuropraxia of the common peroneal nerve; all of these healed completely with appropriate treatment. Therefore we recommend that meniscus repair should be undertaken if a clinical case for meniscal preservation exists, even if the appearances are not those classically recommended. PMID- 12649022 TI - Current status of meniscal transplantation. AB - The current status of meniscal transplantation is reviewed. Results of 36 meniscal transplants carried out by the authors are also presented. PMID- 12649024 TI - Finite element analysis of the meniscus: the influence of geometry and material properties on its behaviour. AB - A finite element model of the knee meniscus was developed to investigate the effects of various geometrical and material properties on the behaviour of the meniscus under compressive load. Factorial methods were used to determine the relative effect of varying the properties by +/-10% of their initial value. It was found that the stresses in the meniscus were more sensitive to geometry (meniscus width and radius of curvature of the femoral surface of the meniscus) than material properties. The model was also used to investigate the effect of incongruency between the radius of curvature of the femur and the femoral surface of the meniscus. It was shown that mismatch between the curvatures of the femur and meniscus has a large effect on the stresses both in the meniscus and in the underlying cartilage. The results from the study have implications for the design and development of meniscal repair devices and replacements. PMID- 12649023 TI - The current state of meniscal allograft transplantation and replacement. AB - This review details current efforts to transplant or to replace a meniscus. Different substitutes for meniscal transplantation or replacement have been used with varying experimental and clinical success. Meniscal transplantation emerges as a useful option for selected patients with a stable knee and appropriate alignment. Some long-term studies of meniscal transplantation prove that cartilage protection is possible. Clear convincing evidence that meniscal transplantation or replacement restores the normal function of the knee joint has not been shown. In keeping with the multiple functions and anatomy of the meniscus experience of meniscal transplantation has shown that many aspects have to be considered to achieve successful results. This offers many possibilities for further scientific investigations. PMID- 12649025 TI - Human meniscal chondrocyte culture and matrix infiltration with specific reference to meniscal generation. PMID- 12649026 TI - Development of an artificial meniscus using polyvinyl alcohol-hydrogel for early return to, and continuance of, athletic life in sportspersons with severe meniscus injury. I: mechanical evaluation. AB - The importance of knee meniscus function is now recognized, and the treatment of meniscus injury has been changing from resection to repair. However, depending on the type of injury, meniscectomy sometimes cannot be avoided. The young athlete might undergo meniscectomy in order to return to sports life as early as possible. However, in such a knee, it is important to anticipate the future problem of degenerative change or osteoarthrosis. In consideration of the prognosis and circumstances in such patients, we have developed an artificial meniscus using polyvinyl alcohol-hydrogel (PVA-H) and performed mechanical tests for compression and stress-relaxation. We found that the human meniscus has unique viscoelastic properties and a high water content. PVA-H showed viscoelastic behaviour similar to that of human meniscus in mechanical tests. These results suggest that an artificial meniscus using PVA-H with a high water content can compensate for meniscal function and might be clinically applicable. PMID- 12649028 TI - The effect of component malalignment on the clinical and radiological outcome of the Kinemax total knee replacement. AB - Component angles of 198 Kinemax total knee replacements were measured from standard short leg radiographs. An ideal tibio-femoral angle of between 4 and 10 degrees of valgus was achieved in 64.6% of patients. After an average follow-up of 6.5 years (range 4.5 to 9.5), there was no significant difference between knees in acceptable and suboptimal alignment in terms of pre- and post-operative knee and function scores and prevalence of radiolucent lines. Varus placement of the tibial component was significantly more common by trainee surgeons (P<0.001). PMID- 12649027 TI - Development of an artificial meniscus using polyvinyl alcohol-hydrogel for early return to, and continuance of, athletic life in sportspersons with severe meniscus injury. II: animal experiments. AB - Due to the prognosis and sporting needs of young athletes who have severe meniscus injury, we have developed an artificial meniscus using polyvinyl alcohol hydrogel (PVA-H). Biomechanical studies have shown that the human meniscus has various mechanical functions; in our mechanical tests, specimens of human meniscus (part I of this study) demonstrated unique viscoelastic properties. Those tests also showed that a high water-content PVA-H has very similar mechanical properties to those of human meniscus and potential as a substitutive meniscus implant. To assess further the use of PVA-H as an artificial meniscus, we have now performed animal experiments using a PVA-H with 90% water content. In rabbits, the lateral meniscus was replaced with an artificial meniscus in one knee of each animal and lateral meniscectomy was performed on the other side as a control. In the knees of meniscectomy group, degenerative changes in articular cartilage progressed with time and osteoarthritis was observed after 1 year. In the knees of the artificial meniscus group, regressive changes were initially observed, but did not progress, and knee joint function was good even after 1 year. Neither wear nor breakage were observed in artificial menisci. These results suggest that an artificial meniscus using a high water-content PVA-H might be clinically applicable. PMID- 12649029 TI - Symmetrical vs asymmetrical total knee replacement--a medium term comparative analysis. AB - Modifications of established implants can result in deleterious effects, as with the Capitol and the matt coated Exeter hips. In 1991 the Kinemax plus modification of the Kinematic knee was introduced in Bristol, the design changes having been made to reduce patello-femoral complications. We carried out a comparative analysis of the prospective recorded data on a consecutive series of 182 total knee replacements (95 Kinematic and 87 Kinemax plus knees) performed between 1991 and 1993. The same instrumentation and surgical technique was used. Since the design had introduced a broader trochlear, offset patella and had changed from an asymmetrical to a symmetrical femoral component, particular attention was paid to tracking and range of movement. All patients were reviewed at 5 years using the Bristol knee score and radiographs. There was no difference in the overall score (both prosthesis scoring 85-86) but the Kinemax plus group with a symmetrical trochlear had a significantly greater improvement in the range of movement (14 degrees as opposed to 4 degrees; P<0.05). In addition, secondary intervention for mal-tracking was significantly less in the Kinemax group. No deleterious effect of the changed geometry was observed. It is concluded that the introduction of a symmetrical femoral component with an offset patella reduced patello-femoral complications without detectable disadvantages. PMID- 12649031 TI - Intraoperative measurements of knee motion in total knee arthroplasty. AB - A photostereometric technology-based knee motion analysis system was developed and intraoperative kinematics during total knee arthroplasty (TKA) was investigated. Ten knees were evaluated and two different types of posterior cruciate-retaining TKA (Genesis-I and Genesis-II) were used. Both TKA showed posterior translation of the medial and lateral femoral condyle with knee flexion. The motion pattern of the estimated contact point of Genesis-II was small initial rollback followed by sliding motion, then rollback again more than 60 degrees. In Genesis-II, the amount of translation of the medial condyle was significantly larger than that of the lateral, suggesting that the lateral condyle acted as a rotational pivot. Our developed knee motion analysis system was non-contact, high resolution and can evaluate both kinematics and estimated contact pattern. The results of this study suggest that intraoperative measurement using this analysis system has advantages for the investigation of in vivo kinematics and contact condition in TKA. PMID- 12649030 TI - Suction technique to improve cement penetration under the tibial component in total knee arthroplasty. AB - Improving the longevity and reliability of cemented total knee arthroplasty remains an important part of orthopaedic research. This paper examines the recently described method of preparation of the cancellous surface of the tibia using suction cannulae in the proximal tibia and stem recess during pulsatile lavage and component fixation. The technique provides a surface clear of debris, fat, blood and marrow, and prevents backflow of blood. A retrospective analysis of post-operative radiographs shows significant improvement in the penetration of cement into the tibial plateau, and improved cement profile towards the periphery. Techniques of stem cementation are also compared, and it is recommended that for this particular design implant, cement should be applied into the cut stem recess in the tibia rather than to the stem of the implant itself. This technique of preparation of the cut surface of the tibia is a simple and, to-date, complication-free method of reliably providing satisfactory cement penetration during total knee arthroplasty. PMID- 12649032 TI - Can knee position save blood following total knee replacement? AB - Previous research showed knee flexion at 70 degrees for 6 h following total knee replacements (TKR) reduced wound drainage by 30%. However, wound drainage may not represent total blood loss and obstruction of the drain in flexion or the relative elevation of the knee could have caused the reduction in drainage. We wished to confirm that this simple, cheap method not only reduced wound drainage but also total haemoglobin loss and determine what part elevation or drain obstruction may play. Sixty consecutive TKR patients were randomly allocated to three groups. Group 1-knee extended and level with bed. Group 2-leg elevated 35 degrees at the hip with knee flexed to 70 degrees for 6 h post-operatively. Group 3-leg elevated 35 degrees at the hip with knee extended for 6 h post-operatively. We assessed wound drainage over 48 h, calculated haemoglobin loss, total blood transfusion, pain, arc of motion, swelling, length of hospital stay and complications. We found knee flexion and knee elevation in extension reduced haemoglobin loss by 25%. Compromise to tissue oxygenation has been reported with knee flexion. We recommend elevation of the leg at 35 degrees from the hip with the knee extended. This offers a simple, safe and effective way to reduce total blood loss by 25%. PMID- 12649033 TI - Evaluation of tibial bone tunnel enlargement using MRI scan cross-sectional area measurement after autologous hamstring tendon ACL replacement. AB - The aim of this study was to evaluate tibial bone tunnel enlargement following four-strand hamstring tendon anterior cruciate ligament (ACL) reconstruction, using a new method of magnetic resonance imaging (MRI) assessment. Correlation with clinical outcome was also examined. In a prospective study following ACL reconstruction, 24 patients underwent MRI and clinical assessment, at a mean follow-up of 6.5 months. A four-strand semitendinosus and gracilis (STG) tendon graft with the polyester Soffix (Surgicraft Ltd) fixation device was placed via an instrument guided tibial tunnel and an over-the-top femoral route. Cross sectional area (CSA) perpendicular to the long axis of the tibial tunnel was calculated digitally at three levels, using MRI. Clinical assessment included physical examination, Lysholm subjective self-evaluation score and KT 2000 arthrometric measurement. A mean CSA tibial tunnel enlargement of 33% was found. The mean side-to-side KT-2000 arthrometric difference was 1.66 mm and the mean Lysholm score was 92. We concluded that this previously unused MRI evaluation using digital direct CSA measurements provides a highly accurate method of tunnel assessment. We found no correlation between tibial tunnel enlargement, knee stability, joint function and patient satisfaction scores in the short term. PMID- 12649034 TI - Intraarticular inflammatory cytokines in acute anterior cruciate ligament injured knee. AB - To understand the dynamics of the intraarticular acute inflammatory phase of an anterior cruciate ligament (ACL) injured knee, we analyzed the level of inflammatory cytokines (TNF-alpha, IL-1beta, IL-6, IL-8, IL-1ra, and IL-10) in joint fluid samples aspirated from 34 knees following an acute ACL injury. The samples were divided into the following five groups according to the duration from injury to aspiration: within 24 h (n=5), 2-3 days (n=14), 4-6 days (n=5), 7 9 days (n=5), 10-14 days (n=4), and 15-21 days (n=3). For comparison, 7 samples were also aspirated from 4 patients with osteoarthritis and 3 with postmenisectomy hydrops (chronic arthritis group). The highest levels of inflammatory cytokines were detected in the ACL-injury group within 24 h of the injury, and the levels decreased thereafter. While there were several patterns of decrease, nearly all of the inflammatory cytokines decreased to the level of that in the chronic arthritis group within 1 week. These dynamics are similar to those reported for inflammatory cytokines in wound fluid during wound healing, and suggest that the intraarticular healing process also progresses in ACL injured knees. PMID- 12649035 TI - The effect of screw length and position on fixation of four-stranded hamstring grafts for anterior cruciate ligament reconstruction. AB - The aim of this study was to examine interference screw fixation of four strand hamstring grafts for anterior cruciate ligament reconstruction in vitro. Bovine tibiae and cannon tendons were used. Screws were introduced from 'outside-in'. The tendons were loaded cyclically to 150 N to simulate walking and 450 N to simulate jogging. Slippage of the tendon from the anchorage was recorded after 100, 300 and 1000 cycles. Cortico-cancellous and cancellous only fixations were compared using a 7-mm screw in an 8-mm tunnel with a 7-mm graft. The effect of screw length was studied by comparing 8 x 25 and 8 x 45 mm screws. Graft slippage with cancellous only fixation was significantly greater than with cortico cancellous screw fixation (P0.105), the longer screws gave more consistent behaviour. We concluded that the screw should be placed so that its head engages the cortex, while increasing screw length within cancellous bone did not significantly improve graft fixation. Cyclical load testing reflects the repetitive forces imposed in the early post operative period following hamstring anterior cruciate ligament reconstruction and is important for the evaluation of graft fixations. PMID- 12649036 TI - Force and repetition in cycling: possible implications for iliotibial band friction syndrome. AB - This study examined force and repetition during simulated distance cycling with regard to how they may possibly influence the on-set of the overuse injury at the knee called iliotibial band friction syndrome (ITBFS). A 3D motion analysis system was used to track lower limb kinematics during cycling. Forces between the pedal and foot were collected using a pressure-instrumented insole that slipped into the shoe. Ten recreational athletes (30.6+/-5.5 years) with no known history of ITBFS participated in the study. Foot-pedal force, knee flexion angle and crank angle were examined as they relate to the causes of ITBFS. Specifically, foot-pedal force, repetition and impingement time were calculated and compared with the same during running. A minimum knee flexion angle of approximately 33 degrees occurred at a crank angle of 170 degrees. The foot-pedal force at this point was 231 N. This minimum knee flexion angle falls near the edge of the impingement zone of the iliotibial band (ITB) and the femoral epicondyle, and is the point at which ITBFS is aggravated causing pain at the knee. The foot-pedal forces during cycling are only 18% of those occurring during running while the ITB is in the impingement zone. Thus, repetition of the knee in the impingement zone during cycling appears to play a more prominent role than force in the on set of ITBFS. The results also suggest that ITBFS may be further aggravated by improper seat position (seat too high), anatomical differences, and training errors while cycling. PMID- 12649037 TI - Neutrophil activity in total knee replacement: implications in preventing post arthroplasty infection. PMID- 12649038 TI - The 5th Bristol unicompartmental arthritis meeting. PMID- 12649040 TI - Cyclooxygenase-2 mediates interleukin-6 upregulation by vomitoxin (deoxynivalenol) in vitro and in vivo. AB - Interleukin-6 (IL-6) is a central mediator of immunotoxicity that is associated with exposure to the trichothecene vomitoxin (VT). The purpose of this investigation was to test the hypothesis that the inducible cyclooxygenase-2 (COX 2) and its metabolites contribute to VT-induced IL-6 upregulation. VT at 100 to 250 ng/ml readily induced COX-2 protein expression in the RAW 264.7 murine macrophage cell line. Superinduction of lipopolysaccharide (LPS)-mediated IL-6 production by VT in these cells was significantly reduced by the COX inhibitors indomethacin and NS-398, whereas the inhibitors did not affect direct induction of IL-6 by LPS alone. Mice that had been gavaged orally with 5 and 25 mg/kg VT exhibited elevated COX-2 mRNA expression in Peyer's patches and spleen with peak induction occurring 2 h after VT exposure. IL-6 mRNA was also induced by VT in vivo, however, peak expression occurred from 2 to 4 h after toxin exposure, suggesting that maximal COX-2 gene upregulation preceded or was concurrent with that for IL-6. Also consistent with a putative contributory role for COX-2 was the finding that both induction of splenic IL-6 mRNA and serum IL-6 by VT were significantly reduced by pretreating mice with the COX inhibitors indomethacin or NS-398. Finally, COX-2 knockout mice showed significantly reduced splenic IL-6 mRNA and serum IL-6 responses to oral VT exposure compared to their parental wild type. Taken together, these in vitro and in vivo data suggest that VT-induced COX 2 gene expression and resultant COX-2 metabolites contributed, in part, to subsequent upregulation of IL-6 gene expression, which has been previously shown to be a hallmark of VT-mediated immunotoxicity. PMID- 12649039 TI - Differential induction of glucocorticoid-dependent apoptosis in murine lymphoid subpopulations in vivo following coexposure to lipopolysaccharide and vomitoxin (deoxynivalenol). AB - Lipopolysaccharide (LPS) and vomitoxin (VT) synergistically induce glucocorticoid mediated apoptotic cell death in lymphoid tissues of the mouse. Based on the known effects of glucocorticoids, it was hypothesized that the combined exposure to LPS and VT targets immature lymphocyte populations. To test this hypothesis, we quantified the effects of VT and LPS on apoptosis induction in T lymphocyte subsets in thymus and B lymphocyte subsets in Peyer's patches and bone marrow. Flow cytometry revealed that a single dose of LPS (0.1 mg/kg body wt ip) together with VT (12.5 mg/kg body wt po) promoted apoptosis of immature (CD4(-)CD8(-), CD4(+)CD8(+)) and mature (CD4(-)CD8(+)) thymocytes at 12 h with a subsequent reduction of these populations being detectable at 24 h. RU 486, a glucocorticoid receptor antagonist, significantly abrogated apoptosis in CD4(-)CD8(-), CD4(+)CD8(+), and CD4(-)CD8(+) subsets and also prevented loss in cell numbers. In Peyer's patches, mature-B lymphocytes (B220(+)IgM(-)IgD(+)) underwent apoptosis and, in bone marrow, pro/pre-B lymphocytes (B220(+)IgM(-)IgD(-)) and mature-B lymphocytes (B220(+)IgM(-)IgD(+)) underwent apoptosis at 12 h after toxin co- exposure. RU 486 blocked LPS + VT-induced apoptosis of the aforementioned subsets in Peyer patches and bone marrow at 12 h. Taken together, these data suggest that LPS can interact with VT in mice to induce the glucocorticoid-driven apoptotic loss of immature thymocytes and cytotoxic T lymphocytes in thymus, mature-B lymphocytes in Peyer's patch, and pro/pre-B lymphocytes and mature-B lymphocytes in bone marrow in mice. PMID- 12649041 TI - Comparison of cis- and trans-crotononitrile effects in the rat reveals specificity in the neurotoxic properties of nitrile isomers. AB - The neurotoxic compound crotononitrile has two isomeric forms, cis and trans. We compared the effects of these two isomers isolated by distillation from the commercially available mixture. Adult male Long-Evans rats were administered vehicle control, cis-crotononitrile (80, 100, and 120 mg/kg/day), or trans crotononitrile (250 mg/kg/day) for 3 days and the changes in corneal transparency and vestibular function were assessed. Surface preparations of the vestibular sensory epithelia and the organ of Corti were examined for hair cell loss by scanning electron microscopy. Concentrations in retina and brain regions of glial fibrillary acidic protein, a marker for reactive gliosis, were also determined in rats exposed to cis-crotononitrile. In a dose-dependent manner, cis crotononitrile induced vestibular dysfunction, corneal opacity, and hair cell loss in both vestibular epithelia and organ of Corti, and gliosis in retina, olfactory bulb, superior colliculus, inferior colliculus, hypothalamus, hippocampus, and cingulate cortex, but not in cerebellum or striatum. This neurotoxic pattern is similar to that caused by 3,3'-iminodipropionitrile and allylnitrile. In contrast, trans-crotononitrile triggered rearing deficits but not vestibular dysfunction, hair cell loss, or corneal opacity. The isomeric specificity of crotononitrile isomers shows that the neurotoxic effects of nitriles depend on strict structural requirements, suggesting that they act through interaction with specific molecular targets. PMID- 12649042 TI - In vitro studies on the chemical reactivity of 2,4-dichlorophenoxyacetyl-S-acyl CoA thioester. AB - 2,4-Dichlorophenoxyacetic acid (2,4-D) is a widely used broadleaf herbicide that has been associated with acute liver toxicity in exposed humans or animals. Chemically reactive metabolites of 2,4-D are proposed as mediators of 2,4-D induced hepatotoxicity. The aim of the present study was to investigate a novel reactive metabolite of 2,4-D, namely 2,4-dichlorophenoxyacetyl-S-acyl-CoA (2,4-D CoA), and to determine its involvement in 2,4-D covalent adduct formation. Thus, incubations of synthetic 2,4-D-CoA (106 microM) with GSH (1 mM) in phosphate buffer (pH 7.4) showed 2,4-D-CoA to be able to transacylate the cysteine sulfhydryl of GSH, resulting in the formation of 2,4-D-S-acyl-glutathione (2,4-D SG) thioester and reaching a concentration of 65 microM after 1 h of incubation. Under similar conditions, 2,4-D-CoA was shown to covalently bind to nucleophilic groups on human serum albumin (HSA, 30 mg/ml), resulting in time-dependent 2,4-D HSA covalent adduct formation that reached a maximum of 440 pmol/mg HSA after 1 h of incubation. In addition to these studies, incubations of [1-(14)C]2,4-D (1 mM) with rat hepatocytes showed a time-dependent covalent binding of 2,4-D to hepatocyte protein. Inhibition of acyl-CoA formation by trimethylacetic acid (2 mM) decreased the amount of covalent binding to protein in rat hepatocytes by 50%. These results indicate that 2,4-D-CoA thioester is a reactive metabolite of 2,4-D that may contribute to 2,4-D-protein adduct formation in vivo and therefore the associated hepatotoxicity. PMID- 12649044 TI - Importance of the kidneys in metabolism of vitamins A1 and A2 and their fatty acyl esters in mink feeding on fish-based diets and exposed to Aroclor 1242. AB - Two-month-old female mink (Mustela vison) were fed diets based on freshwater smelt (Osmerus eperlanus), Baltic herring (Clupea harengus membras), or North Atlantic herring (Clupea harengus harengus) for 21 weeks. Half of the smelt-fed mink were exposed to the commercial polychlorinated biphenyl (PCB) preparation Aroclor 1242 (1 mg/day) in the feed. Retinol (vitamin A(1)), 3,4-didehydroretinol (vitamin A(2)), their fatty acyl esters, and vitamin E were studied in the kidneys by reversed-phase high-performance liquid chromatography. Exposure to Aroclor 1242 resulted a significant decrease in the alcoholic and esterified vitamin A(2). Levels of the A(1) analogs did not change due to the PCBs and were the same in mink fed either smelt or Baltic herring. In mink fed very fatty Atlantic herring, the renal levels of vitamin A(1) and E were significantly lower than in the other mink and apparently consumed by lipid peroxidation. The vitamins were located mainly in the cortex, and the fatty acyl esters showed a fatty acid composition that differed from those in liver and plasma. In the kidneys of the smelt-fed mink (control or Aroclor-exposed) the ratio of vitamin A(2) to A(1) was much lower than the ratios in the liver or plasma, suggesting inefficient uptake of A(2) in the kidneys. In the PCB-exposed mink, in which vitamin losses are increased, tissue levels of vitamin A(2) may be more difficult to maintain than levels of vitamin A(1). PMID- 12649043 TI - Competitive interactions of chlorinated phenol compounds with 3,3',5 triiodothyronine binding to transthyretin: detection of possible thyroid disrupting chemicals in environmental waste water. AB - Chlorinated phenol compounds, such as the chlorinated derivatives of bisphenol A, have been detected in effluents from paper manufacturing plants. We investigated the effects of bisphenol A, nonylphenol, and their seven chlorinated derivatives on 3,3',5-[(125)I]triiodothyronine ([(125)I]T(3)) binding to purified chicken and bullfrog transthyretin (cTTR and bTTR) and to the ligand-binding domains of chicken and bullfrog thyroid hormone receptor beta (cTR LBD and bTR LBD). The concentrations at which the chlorinated derivatives displaced [(125)I]T(3) from TTR were 10-10(3) times less than those of their parent molecules. 2,6-Dichloro-4 nonylphenol and 3,3',5-trichlorobisphenol A were the most potent competitors of T(3) binding to cTTR and to bTTR, respectively. The interactions of the chlorinated derivatives with the cTR and the bTR LBDs were weaker than those of the chlorinated derivatives with cTTR and bTTR. Chlorinated derivatives with a greater degree of chlorination were more efficient competitors of T(3) binding to TTR and TR. A structure-activity relationship between the phenol compounds and TTR (TTR assay) and TR (TR assay) was established. Structures with chlorine in either ortho position or both ortho positions, with respect to the hydroxy group, were more efficient competitors. Chemicals that interacted with bTTR, cTTR, and Japanese quail TTR were detected in water downstream of effluents from paper manufacturing plants using the TTR assay. As some of the chlorinated bisphenols and nonylphenols were potent competitors of T(3) binding to TTRs, the TTR assay could be applied as primary screening for possible thyroid-disrupting chemicals in environmental waste water. PMID- 12649045 TI - Dose-response analysis of short-term effects of 2,3,7,8-tetrachlorodibenzo-p dioxin in three differentially susceptible rat lines. AB - Line A, B, and C rats were selectively bred from TCDD-resistant Han/Wistar (Kuopio; H/W) and TCDD-sensitive Long-Evans (Turku/AB; L-E) rats. Line A rats are the most resistant to TCDD acute lethality followed by line B and line C rats. The resistance in line A rats is associated with a mutated H/W-type aryl hydrocarbon receptor (Ahr) allele (Ahr(hw)) and in line B rats the resistance is associated with an allele of an unknown gene B (B(hw)), while line C rats are almost as sensitive to TCDD as L-E rats. The dose-responses of characteristic short-term effects (day 8 postexposure) of TCDD were used to evaluate the efficacy (magnitude of effect) and potency relationships between these lines. Line A rats showed similar efficacies as line C (line A:line C efficacy ratio more than 0.7) for thymus weight, EROD activity, and incisor tooth defects. In contrast, efficacies in line A were decreased (efficacy ratios 0.19-0.37) for body weight change, serum bilirubin, and FFA levels, and serum ASAT activity. For most endpoints the efficacies in line B rats seem to be lower than in line C rats. The potencies were close to each other in line A and B rats, but somewhat lower than in line C rats. The results support our previous concept of two different AHR-mediated signaling pathways leading to dioxin type I and type II endpoints. Rats with the Ahr(hw/hw) genotype show a markedly decreased efficacy for type II endpoints, but B(hw) allele had only a minor effect on efficacies for most endpoints. Both H/W-type resistance alleles also decreased the potency of TCDD. However, the potency differences in short-term toxicity seem not to explain, at least alone, the differences seen in acute lethality among the rat lines. PMID- 12649046 TI - National trends in screening, prevalence, and treatment of cardiovascular risk factors. AB - BACKGROUND: Few studies have documented national trends in screening, awareness, and treatment of cardiovascular risk factors. We evaluated trends in screening, prevalence, and treatment of hypertension, hypercholesterolemia, and smoking. METHODS: Data were analyzed from the 1984-1998 Behavioral Risk Factor Surveillance System, a series of yearly cross-sectional population-based surveys of U.S. adults. Unadjusted and adjusted time trends (age-, gender-, ethnicity-, education-, and income-adjusted) in screening, prevalence, and treatment were evaluated. RESULTS: From 1984 to 1998, a larger proportion of U.S. adults were older, more educated, richer, and Hispanic. Hypertension screening was >97% (1988 1998), prevalence ranged from 21 to 24% (1984-1998), and approximately 58% (1984 1992) were prescribed blood-pressure-lowering medications. Hypercholesterolemia screening increased from 47 to 67% (1987-1998), prevalence from 18 to 31% (1987 1998), and cholesterol-lowering prescriptions from 22 to 25% (1988-1990). Smoking prevalence remained around 28% (1984-1998), while quit attempts declined from 63 to 47% (1990-1998). CONCLUSIONS: Although screening for hypertension and hypercholesterolemia has increased, a substantial proportion of cases were not being prescribed medications. While the prevalence of smoking remains constant, quit attempts have fallen. Continuing challenges for cardiovascular disease prevention include identification of individuals with hypercholesterolemia, appropriate prescription (initiation and/or maintenance) of antihypertensive and lipid-lowering medications, and intensifying smoking cessation efforts. PMID- 12649047 TI - Awareness of hypertension and dyslipidemia in a semirural population of young adults: the Bogalusa Heart Study. AB - BACKGROUND: To assess the awareness of hypertension and dyslipidemia in a semirural population of young adults. METHODS: A cohort of 5,707 individuals was surveyed by a questionnaire. The awareness analysis was done on 1,454 subjects screened for cardiovascular risk factors 5 years earlier. RESULTS: Among the respondents (n = 3,699, 65%), the prevalence of hypertension among blacks and whites was 11% and 7.3% (P < 0.0001), respectively; dyslipidemia, 4.7% and 5.8% (P = 0.27). Results from the screening showed a prevalence of 11.4% in blacks versus 6.0% in whites (P < 0.0001) for hypertension; 14.2% versus 17.9% (P = 0.12) for dyslipidemia. Males had a higher prevalence of dyslipidemia than females (24% vs. 12%, P = 0.001). Five years later, among those informed of their hypertension, males were more likely to be aware of their hypertension than females [odds ratio (95% confidence interval) = 5.0 (1.4-17.5)]. Increasing age [1.04 (1.0-1.1)], positive parental history of coronary heart disease [2.6 (1.5 4.3)], and higher education level [2.1 (1.2-33.6)] were associated with the awareness of dyslipidemia. Awareness of the condition increased the proportion of subjects receiving treatment for hypertension, but did not change the proportion receiving treatment for dyslipidemia. CONCLUSIONS: These results underscore the need for cardiovascular health education efforts in this population group. PMID- 12649048 TI - Cost-effectiveness of primary prevention of coronary heart disease through risk factor intervention in 60-year-old men from the county of Stockholm--a stochastic model of exercise and dietary advice. AB - BACKGROUND: Recent screenings show a high prevalence of cardiovascular risk factors in the county of Stockholm. Primary prevention may be a way to lower the risk burden of coronary heart disease, but we must establish that preventive programs are cost-effective. METHODS: Through the use of a stochastic Markov model, which predicts reduction in coronary heart disease events based on risk factor reductions, this study evaluates the results of a previous controlled trial in middle-aged men comparing dietary advice, exercise, and the combination of both applied to an observed cohort of 60-year-old men in the county of Stockholm. RESULTS: The model predicts lower costs and higher effectiveness for dietary advice compared to the alternatives. Assuming a declining effect of the intervention, dietary advice saves 0.0228 life-years compared to no intervention. If no decline is assumed, the corresponding figure is 0.0997 life-years. From the societal perspective, the added costs are 2,892 Swedish Kronor (SEK) and 14,106 SEK for the two modeling assumptions, resulting in a cost-effectiveness of 127,065 SEK per life-year gained (LYG) and 141,555 SEK/LYG. These figures are below what is generally thought of as cost-effective. CONCLUSION: Based on the model, dietary advice appears to be the most cost-effective of the studied interventions. PMID- 12649049 TI - The D-Net diabetes self-management program: long-term implementation, outcomes, and generalization results. AB - BACKGROUND: A prerequisite to translating research findings into practice is information on consistency of implementation, maintenance of results, and generalization of effects. This follow-up report is one of the few experimental studies to provide such information on Internet-based health education. METHODS: We present follow-up data 10 months following randomization on the "Diabetes Network (D-Net)" Internet-based self-management project, a randomized trial evaluating the incremental effects of adding (1) tailored self-management training or (2) peer support components to a basic Internet-based, information focused comparison intervention. Participants were 320 adult type 2 diabetes patients from participating primary care offices, mean age 59 (SD = 9.2), who were relatively novice Internet users. RESULTS: All intervention components were consistently implemented by staff, but participant website usage decreased over time. All conditions were significantly improved from baseline on behavioral, psychosocial, and some biological outcomes; and there were few differences between conditions. Results were robust across on-line coaches, patient characteristics, and participating clinics. CONCLUSIONS: The basic D-Net intervention was implemented well and improvements were observed across a variety of patients, interventionists, and clinics. There were, however, difficulties in maintaining usage over time and additions of tailored self-management and peer support components generally did not significantly improve results. PMID- 12649050 TI - Relative validity of a diet history interview in an intervention trial manipulating dietary fat in the management of Type II diabetes mellitus. AB - BACKGROUND: This study assesses the relative validity of a diet history interview in 56 free-living individuals with Type II diabetes mellitus. METHODS: The diet history interview was compared to a 3-day food record in a 1-year dietary intervention trial. The plausibility of energy intake data was examined throughout the trial. Paired data were compared for differences and the presence of systematic error was determined by Bland Altman analysis. Changes in accuracy and responsiveness were assessed over time. RESULTS: The proportion of underreporters was larger in the diet history at baseline. Underrecording with the food record was more common in subjects with BMIs > 30 kg/m(2). There was no difference between paired dietary data from the two methods; however, data on fatty acids failed to correlate. These correlations improved when outliers were removed. There was no evidence of a relationship between bias and mean intake of dietary variables. Accuracy of diet history measurement did not change during the trial for energy or macronutrients, but data on protein and monounsaturated fat were both affected by BMI. The diet history was more responsive than the food record to changes in monounsaturated fatty acid intake after 3 months, but this changed at the end of the trial. CONCLUSION: The diet history provided good estimates of energy and macronutrient intakes in a sample group with Type II diabetes mellitus. However, energy intake data revealed a high prevalence of underreporting especially in people with higher BMIs. PMID- 12649052 TI - Brief, validated survey instruments for the measurement of fruit and vegetable intakes in adults: a review. AB - BACKGROUND: Brief surveys measuring fruit and vegetable intakes of populations have been used to monitor local and national trends in fruit and vegetable consumption over time, and to evaluate interventions to promote fruit and vegetable consumption for the primary prevention of disease. To date, brief validated survey instruments measuring self-reported fruit and vegetable consumption levels have not been systematically reviewed. METHODS: MEDLINE search for papers describing validated survey instruments with 16 or fewer fruit and vegetable items. RESULTS: Ten survey instruments with total numbers of 6 to 16 items met our search criteria. In comparisons with in-depth dietary assessment methods, survey instruments with relatively greater numbers of fruit and vegetable items, and with questions on portion sizes and mixed vegetable dishes, were characterized by higher Pearson and/or Spearman rank correlation coefficients for fruit and vegetable intakes and by closer estimations of mean/median total fruit and vegetable intakes. CONCLUSIONS: This review suggests that the inclusion of a moderate number of representative fruit and vegetable items, and the review of questions on portion size and the consumption of mixed vegetable dishes, may enhance the validity of brief fruit and vegetable instruments. PMID- 12649051 TI - Feedback strategies to raise awareness of personal dietary intake: results of a randomized controlled trial. AB - BACKGROUND: Feedback is a strategy that can be used to influence awareness of dietary habits. Feedback was applied in an interactive computer-tailored intervention and in printed self-test forms. METHODS: A randomized controlled trial with a 3 (study groups) x 2 (higher vs. medium or lower educational level) design was conducted. Adult subjects (N = 304) were randomly assigned to a feedback group or the control group. Immediate impact on realism of self-rated intake levels of fat, fruit, and vegetables were tested, as were intentions to change. RESULTS: Self-rated fat intake compared to others was more realistic among respondents with a medium or lower educational level in the tailored intervention group. Self-rated fruit intake compared to others was more realistic in the tailored intervention group. Self-rated fat intake was more realistic in the tailored intervention than the self-test group. Intention to reduce fat consumption was greater in the tailored intervention group. Intention to eat more vegetables was greater in the tailored than in the self-test group. Subjects rated the tailored intervention as more effective, more personally relevant, more individualized, and providing more new information. CONCLUSIONS: Only the tailored intervention had an immediate impact on awareness and dietary change intention and was appreciated better than both other interventions. PMID- 12649053 TI - Trends in cigarette smoking in the German centers of the European Prospective Investigation into Cancer and Nutrition (EPIC): the influence of the educational level. AB - BACKGROUND: Several studies in Germany and other European countries have already shown smoking prevalence to be related to education. This study was aimed to investigate time trends in smoking habits in the German cohorts Heidelberg and Potsdam of the European Prospective Investigation into Cancer and Nutrition (EPIC) according to sex, birth cohort, and level of education. METHODS: Within EPIC, 25,546 and 27,548 participants were recruited in Heidelberg and Potsdam, respectively. Data on smoking were collected by means of a computer-guided interview during the baseline examination between 1994 and 1998. For each birth cohort smoking prevalence and mean number of cigarettes smoked per day at different ages were calculated. Odds ratios and 95% confidence interval for associations between smoking prevalence and educational level were computed by using logistic regression. RESULTS: Smoking prevalence was higher among men than among women, with a smaller difference in younger birth cohorts. Between 1950 and 1960, smoking prevalence among women in the Heidelberg cohort rose sharply (from 12.8% to 51.8% in the least educated group). This strong increase was delayed by 10 years in the Potsdam cohort. Men and women in Heidelberg smoked more cigarettes per day than their counterparts in Potsdam, but in both study centers less educated subjects smoked more than subjects with a higher education. CONCLUSIONS: Smoking patterns in the Potsdam and Heidelberg cohorts are quite similar with respect to prevalence and years of lifetime smoking. Since an increasing difference between smoking prevalence of less and high educated individuals is observable, programs on smoking cessation should especially concentrate on persons of lower educational level. PMID- 12649054 TI - Rapid progression to regular cigarette smoking among nonsmoking adolescents: interactions with gender and ethnicity. AB - BACKGROUND: While the correlates and predictors associated with transition through the stages of smoking acquisition have received substantial attention, the identification of factors associated with the time course of progression remain generally unexplored. METHODS: We identified adolescents escalating from nonsmoking to regular smoking in 1 year as rapid progressors (n = 98) and those escalating from nonsmoking to experimental smoking in 1 year as slow progressors (n = 555). We entered demographic and psychosocial variables, initial reactions to smoking variables, and their interactions with gender and ethnicity into a logistic regression analysis comparing rapid progressors to slow progressors. RESULTS: At baseline, rapid progressors reported more friends who smoked regularly. Girls who reported more concern with dieting and less social success were more likely to progress to regular smoking within 1 year; these variables did not influence progression in boys. Those who associated positive feelings with their initial smoking experience were more likely to progress to regular smoking. Differences between white and African American youth emphasize the importance of physiological interpretations of initial smoking experiences. CONCLUSIONS: Our findings suggest that even as nonsmokers, adolescents likely to progress quickly to regular smoking can be identified. Researchers designing prevention programs should take these factors into account. PMID- 12649055 TI - Impact of patient characteristics on physician's smoking cessation strategies. AB - BACKGROUND: The effectiveness of smoking cessation interventions may be influenced by a variety of patient characteristics, including level of nicotine addiction and readiness to change. We conducted this study to examine the relationship between these characteristics and the frequency of physician initiated smoking cessation interventions. METHODS: We identified smokers seen during office visits to 1 of 38 primary care physicians in rural Kansas. Trained students observed the frequency and nature of doctor-patient discussions related to tobacco. Telephone surveys were conducted with these patients 1 to 3 days after the office visit. RESULTS: We completed observations and telephone surveys on 259 smokers. Tobacco-related discussions occurred during 66% of doctor-patient encounters. Although discussions overall were unrelated to a patient's readiness to quit, specific assistance with smoking cessation was offered less often to precontemplators (15%) than to contemplators (31%) or those preparing to quit (37%) (P < 0.05). While bupropion was discussed with 23% of smokers, nicotine replacement therapy was discussed with 12% and was unrelated to markers of nicotine addiction. CONCLUSIONS: Current efforts to promote smoking cessation are only marginally related to patient characteristics. Doctors are missing many opportunities to effectively intervene with patients who are contemplating smoking cessation or preparing to quit. PMID- 12649056 TI - Changing multiple health behaviors: smoking and exercise. AB - BACKGROUND: Previous stage of change research examining health behaviors has tended to examine one behavior at a time. However, one recent study by King et al. (1996) examined the relationship between smoking and exercise across cognitive-behavioral mediators (i.e., decisional balance and self-efficacy) shown to be important in predicting readiness to change. In this study, we seek to replicate the study of King et al. (1996) in a low-income sample, the majority of whom are women, with at least one chronic illness who are attending primary care clinics. METHODS: Data were obtained from 270 adult outpatients attending four public primary care clinics in Louisiana. RESULTS: Smoking and exercise stage of change were not related. Significant relationships existed between the cognitive variables of smoking and exercise. No significant differences existed within exercise stage of change on the cognitive variables of smoking, and vice versa, no significant differences were noted within smoking stage of change on the cognitive variables of exercise. CONCLUSIONS: Smoking and exercise appear to be specific health behaviors that are independent constructs in this particular sample. However, caution should be taken when interpreting the findings since 75% of the sample had at least one chronic illness. PMID- 12649057 TI - Rationale, design, and baseline data for Commit to Quit II: an evaluation of the efficacy of moderate-intensity physical activity as an aid to smoking cessation in women. AB - BACKGROUND: Commit to Quit II is a 4-year randomized controlled trial comparing the efficacy of a cognitive-behavioral smoking cessation treatment plus moderate intensity physical activity with the same cessation treatment plus contact control. METHODS: Sedentary women smokers (n = 217) were randomized to receive 8 weeks of treatment followed by 12 months of follow-up. This article outlines the study design, presents baseline data about the sample, and compares the sample to national samples and to our previous study examining vigorous-intensity exercise as an aid to smoking cessation. RESULTS: Married and white participants reported significantly higher levels of nicotine dependence than unmarried and minority participants. Higher levels of nicotine dependence were also significantly related to lower smoking cessation self-efficacy and higher levels of self reported depression, anxiety, and perceived stress. Additionally, participants smoked significantly more cigarettes (mean 20.6) than a national sample of female smokers (mean 16.1). On average, participants were significantly older, weighed significantly more, and scored significantly higher on a measure of anxiety than participants in our previous trial. CONCLUSIONS: Our sample consisted of women who were heavier smokers than national samples seeking treatment. It remains to be determined how this will impact their ability to attain cessation in the present study. PMID- 12649058 TI - Can we skill and activate children through primary school physical education lessons? "Move it Groove it"--a collaborative health promotion intervention. AB - BACKGROUND: Physical education (PE) lessons are an ideal setting to improve child fundamental movement skills (FMSs) and increase physical activity (PA) for optimal health. Despite this, few studies have assessed the potential to do both simultaneously. The "Move It Groove It" primary school intervention in New South Wales, Australia, had this opportunity. METHODS: A whole school approach to implementation included establishment of school project teams, a teacher "buddy" system, project Web site, teacher training workshops, and small grants for equipment. The quasi-experimental evaluation involved 1,045 year 3 and 4 children (aged 7 to 10 years) in nine intervention and nine control rural primary schools (53% boys/47% girls). It utilised pre- and postobservational surveys of (1) mastery or near mastery levels for each of eight FMSs, (2) proportion of PE lesson time spent in moderate to vigorous PA (MVPA) and vigorous PA (VPA), and (3) teacher- and lesson-related contextual covariates. Data were analysed by hierarchical logistic multiple regression. RESULTS: For FMSs, overall mastery or near mastery level at baseline was 47% ranging from 22.7% for the overarm throw among girls to 75.4% for the static balance among boys. The intervention delivered substantial improvements in every FMS for both genders ranging from 7.2% to 25.7% (13 of 16 comparisons were significant). For PA level, mean MVPA at baseline was 34.7%. Baseline MVPA for boys was 38.7% and for girls was 33.2%. The intervention was associated with a nonsignificant 4.5% increase in MVPA and a significant 3.0% increase in VPA. This translates to a gain of <1 minute of MVPA per average 21-minute lesson. CONCLUSIONS: This is the first study to show that by modifying existing PE lessons, significant improvements in FMS mastery can be gained without adversely affecting children's MVPA and VPA. To increase PA levels, we recommend increasing the number of PE lessons per week. PMID- 12649059 TI - Asymptomatic sexually transmitted diseases: the case for screening. AB - BACKGROUND: The United States has the highest prevalence of sexually transmitted diseases in the developed world. Control strategies should address the most frequent reasons why curable sexually transmitted diseases are not treated. METHODS: We approached 1,631 persons ages 18-29 in various sites and offered them screening for gonorrhea and chlamydial infection and surveys regarding past genitourinary symptoms. For those with past symptoms we abstracted medical records or conducted additional interviews. From these data we estimated the total number of persons who had gonorrhea or chlamydial infections in the previous year, the proportion treated, and the primary reasons for nontreatment. RESULTS: The prevalence of gonorrhea was 2.3% and that of chlamydial infection was 10.1%. We estimate that 45 and 77% of all cases of gonorrhea and chlamydial infection, respectively, were never symptomatic and that 86 and 95% of untreated cases of gonorrhea and chlamydial infection, respectively, were untreated because they were never symptomatic. The remaining 14 and 5% of untreated cases of gonorrhea and chlamydia, respectively, were not treated because persons did not receive medical care for symptoms. CONCLUSIONS: The primary reason that gonorrhea and chlamydial infections are untreated is that infected persons never have symptoms. The most effective method to control these sexually transmitted diseases is routine screening at high-volume sites. PMID- 12649060 TI - Implementing a seventh grade vaccination law: school factors associated with completion of required immunizations. AB - OBJECTIVES: We investigated school factors associated with successful implementation of a seventh grade vaccination requirement. METHODS: The proportion of students vaccinated with hepatitis B vaccine and measles containing vaccine was determined from records of schools in San Diego County, California. A school survey identified compliance strategies. Analysis identified factors associated with coverage. RESULTS: In October 1999, 67.2% of 38,875 students had received the required vaccine doses. Of 315 schools, coverage was less than 40% in 60 schools and exceeded 80% in 111 schools. Factors associated with high coverage included private schools, early and frequent notice to parents, and, for public schools, higher overall socioeconomic status of students. CONCLUSIONS: In preparation for a middle school vaccination requirement, early and frequent notification of parents improves coverage. Schools with a high percentage of low socioeconomic status students may require extra resources to support implementation. PMID- 12649062 TI - Selective DNA recognition by the androgen receptor as a mechanism for hormone specific regulation of gene expression. AB - The androgen receptor (AR) is a member of the highly conserved group of the class I steroid hormone receptors, a subgroup of the nuclear receptor superfamily of ligand-induced transcription factors. All class I receptors influence the expression of their target genes by binding three-nucleotide spaced partial palindromic repeats of the core 5'-TGTTCT-3' motif. The implication that all class I receptors activate transcription by binding similar DNA motifs, poses the problem of how the in vivo steroid-specificity of transcriptional control is achieved. The AR, however, is able to interact with DNA motifs that are divergent from the classical hormone response elements. We will describe this AR-specific DNA interaction in the context of the general mechanisms that dictate the sequence-specificity of DNA-binding and dimerization of the nuclear receptors. The androgen receptor is the only steroid hormone receptor that is able to interact with response elements that are essentially arranged as a direct repeat of the 5'-TGTTCT-3' monomer binding element. We propose that the DNA-binding domain of the AR can interact with these androgen-specific response elements in a head-to-tail conformation, similar to many other nuclear hormone receptors. The fact that subtle differences in the sequence of response elements can dictate androgen-specific responses is a new and intriguing finding. It creates new possibilities in the research on hormone-selective action and provides a new angle in the search for selective ligands or co-factors that might influence androgen receptor action via either type of DNA motif. PMID- 12649061 TI - Replacement therapy in Mucopolysaccharidosis type VI: advantages of early onset of therapy. AB - This study evaluates the immunological response following weekly 2h infusions of recombinant human N-acetylgalactosamine 4-sulfatase (rh4S) in Mucopolysaccharidosis VI (MPS VI) cats. The results of three trials (Trial "A": 9 month duration with onset at 3-5 months of age, n = 5; and Trials "B" and "C": 6 month duration starting at birth, n = 9) were compared. No detrimental effects were noted throughout Trials B and C. Temporary hypersensitivity reactions (e.g., vomiting, diarrhoea) occurred in four cats in Trial A and were alleviated by increasing the dose of antihistamine premedication and the duration of infusion. All cats in Trial A developed antibodies to rh4S (range of final titres: 1041 134,931). All cats treated from birth showed negligible titres (range: < 50-598). In vitro inhibition of rh4S activity (up to 47%) was demonstrated with plasma from four cats with elevated titres. Significant reduction of urinary glycosaminoglycan concentration in all cats indicated the ability of rh4S to metabolize stored substrates regardless of the presence of circulating antibodies. Similarly, lysosomal storage in reticuloendothelial cells and fibroblasts of kidney interstistium, dura and skin was reduced in all cats irrespective of their antibody titre although cats with elevated titre had less beneficial effect on cardiovascular tissues (aorta smooth muscle cells, heart valve fibroblasts). Overall improvement in the disease condition (at physical, neurological, and skeletal levels) was most pronounced for cats treated from birth compared with cats treated at a later age. PMID- 12649063 TI - Impaired mitochondrial pyruvate importation in a patient and a fetus at risk. AB - The patient was the first child of healthy consanguineous parents. She presented at birth with hypotonia, mild facial dysmorphism, periventricular cysts, marked metabolic acidosis, hyperlactacidemia with normal lactate/pyruvate molar ratios, normoglycemia, and normal ammonia. Hyperlactacidemia was severe (5-14 mmol/l) and not corrected with bicarbonate, thiamine (10 mg/d), 2-chloropropionate (100 mg/kg/d) and a ketogenic diet. Pyruvate dehydrogenase (PDHC) activity was normal in lymphocytes and fibroblasts. Functional assays were performed in digitonin permeabilized fibroblasts to measure oxidation rates from radiolabeled pyruvate and malate. The production of [14C]acetylcarnitine or [14C]citric cycle intermediates derived from [2-14C]pyruvate as well as the release of 14CO(2) from [1-14C]pyruvate was severely impaired, whereas decarboxylation of [U-14C]malate was normal. With increasing concentrations of [1-14C]pyruvate, the patient's fibroblasts behave like control fibroblasts incubated in the presence of alpha cyano-4-hydroxycinnamate, a specific inhibitor of mitochondrial pyruvate uptake: a progressive increase in 14CO(2) production was observed, likely due to passive diffusion of [1-14C]pyruvate through the mitochondrial membranes. Our results are consistent with a defect of mitochondrial pyruvate transport in the patient. Mutational analysis was precluded as the cDNA sequence of the pyruvate carrier has not been identified as yet in any organism. An affected fetus was recognized in a subsequent dichorionic twin pregnancy using the coupled assay measuring [2 14C]pyruvate oxidation rates on digitonin-permeabilized trophoblasts. After selective feticide, the pregnancy was uncomplicated with delivery at 37w of a healthy female, who is currently 2-month old. PMID- 12649064 TI - Determination of monosaccharides and disaccharides in mucopolysaccharidoses patients by electrospray ionisation mass spectrometry. AB - The mucopolysaccharidoses are a group of lysosomal storage disorders characterised by the storage of glycosaminoglycans. With the exception of Hunters syndrome (MPS II), which is X-linked, they are autosomal recessively inherited resulting in a defect in any one of 10 lysosomal enzymes needed to catabolise glycosaminoglycans. The type and size of the glycosaminoglycans stored in lysosomes are determined by the particular enzyme deficiency. These glycosaminoglycan elevations are subsequently observed in tissue, circulation, and urine. A method has been developed for the derivatisation and quantification of sulfated N-acetylhexosamine-containing mono- and disaccharides from patient samples by electrospray ionisation tandem mass spectrometry. Urine from most mucopolysaccharidoses types had significant increases in di- and monosulfated N acetylhexosamines (GalNAc4,6S, GalNAc6S, GalNAc4S, or GlcNAc6S) and monosulfated N-acetylhexosamine-uronic acid disaccharides (GalNAc6S-UA, GalNAc4S-UA, or GlcNAc6S-UA). Analysis of plasma and dried blood spots on filter paper collected from mucopolysaccharidoses patients showed elevations of total monosulfated N acetylhexosamines but less than that seen in urine. Urine samples from bone marrow transplant recipients, mucopolysaccharidosis IVA and mucopolysaccharidosis VI patients, showed decreases in HexNAcS, HexNAcS(2)/GalNAc4,6S, and HexNAcS-UA post-transplant. This decrease correlated with clinical improvement to levels comparable with those identified in patients with less severe phenotypes. These metabolic markers therefore have potential applications in diagnosis, phenotype prediction and monitoring of current and future therapies, particularly for the mucopolysaccharidosis IIID, IVA, VI, and multiple sulfatase deficiency. This paper reports a sensitive and simple method for the measurement of sulfated N acetylhexosamines and sulfated disaccharides shown to be elevated in some mucopolysaccharidosis and multiple sulfatase deficient patients. PMID- 12649065 TI - DHPLC mutation analysis of phenylketonuria. AB - Denaturing high-performance liquid chromatography (DHPLC) is a sensitive and fast method for the detection of mutations which has been successfully used for mutation screening in several disease-related genes. Phenylketonuria (PKU, OMIM* 261600; McKusick 1986) is one of the most common autosomal recessive disorders in Europe. Mutations in the PAH gene mainly involve point mutations. In this study we report the successful use of DHPLC to analyse rapidly the complete coding sequence of the PAH gene in a total of 125 unrelated patients with PKU. PMID- 12649066 TI - Gene-gene interaction between the cystathionine beta-synthase 31 base pair variable number of tandem repeats and the methylenetetrahydrofolate reductase 677C > T polymorphism on homocysteine levels and risk for neural tube defects. AB - INTRODUCTION: Most studies showed that mothers of children with NTD have elevated homocysteine levels pointing to a disturbed homocysteine metabolism as a risk factor for NTD. Folate lowers homocysteine levels by remethylation of homocysteine to methionine. Homocysteine can be irreversibly converted to cystathionine by the vitamin B6-dependent enzyme CBS. Recently, our group showed that a 31 bp VNTR in the CBS gene was associated with decreased CBS activity and increased tHcy levels after methionine loading in a CVD population. AIM: The aim of our study was to investigate whether this VNTR influences tHcy levels and risk for NTD. In addition, we assessed the role of vitamin B6 as an effect modifier in this possible interaction. We examined possible gene-gene interaction with the MTHFR 677C > T polymorphism. We screened genomic DNA of 88 NBD patients, 100 mothers, 88 fathers, and 505 controls for this CBS 31 bp VNTR. RESULTS: In this study population five different alleles with 16,17, 18, 19, and 21 times the 31 bp repeat were observed that constituted 10 different genotypes. The most common 18/18 VNTR genotype was associated with higher tHcy levels compared with the 17/18 and 18/19 VNTR genotypes. Vitamin B6 levels did not influence this association. In addition, no association with risk for NTD was found. Combination of the CBS VNTR with the MTHFR 677C > T polymorphism revealed an additional increase in homocysteine levels in 18-18 individuals compared with 17-18 peers within subjects homozygous mutant for the MTHFR 677C > T polymorphism. CONCLUSIONS: The present study indicates that the number of 31 bp repeat elements in the CBS gene influences tHcy levels. This VNTR seems not to be associated with an increased risk for NTD. PMID- 12649067 TI - Homocysteine remethylation enzyme polymorphisms and increased risks for neural tube defects. AB - Folic acid supplementation can effectively reduce the risk of neural tube defects (NTDs); however, the mechanism underlying this beneficial effect remains unclear. Recent evidence suggests that certain folate pathway genes, as well as those related to homocysteine metabolism might be contributing to this effect. The purpose of this study is to investigate whether gene polymorphisms of methionine synthase (MTR) and methionine synthase reductase (MTRR) are involved in the risk for NTDs, specifically spina bifida. We detected MTR A2756G and MTRR A66G polymorphisms using PCR-RFLP analysis in a group of NTD infants, their mothers and normal controls. We found that infants with the MTRR mutant genotype had a 2.6-fold higher risk of NTDs when compared to the AA genotype (OR = 2.6, 95%CI = 1.3-5.3). Mothers with the MTRR mutant genotype also had a 1.9-fold higher risk of having an NTD baby compared to AA genotype (OR = 1.9, 95%CI = 1.1-3.1). Infants who carry mutant alleles for both MTRR and MTR had exceptionally elevated NTD risks, with odds ratios of 5.1 compared to infants with the wild type genotype at both loci (AA + AA) (OR = 5.1, 95%CI = 1.7-15.4). A comparable result was observed in the mothers of NTD cases (OR = 2.1, 95%CI = 1.0-4.7). Our results indicate that MTRR and MTR genes may interact to increase the infants' NTD risks. These results did not appear to be influenced by maternal periconceptional folic acid intake. However,the sample size of this study was limited, and a larger population study is needed to pursue these initial observations. PMID- 12649068 TI - New mutations in two Dutch patients with early infantile galactosialidosis. AB - Galactosialidosis is an autosomal recessive lysosomal storage disease caused by a combined deficiency of lysosomal beta-galactosidase and neuraminidase as a result of a primary defect in the protective protein/cathepsin A (PPCA). We report the first 2 Dutch cases of early infantile galactosialidosis, both presenting with neonatal ascites. The defect was identified in urine, leukocytes, and fibroblasts. Residual activity was determined with a modified assay for cathepsin A and was <5% in leukocytes and <1% in fibroblasts. Histological examination of the placenta in case 1 showed extensive vacuolization in all cell types. Northern blot analysis of RNA isolated from the patients' cultured fibroblasts showed substantially decreased levels of the PPCA transcript, which nevertheless had the correct size of 2 kb. Mutation analysis of both mRNA and genomic DNA from the patients identified two novel mutations in the PPCA locus. Case 1 was a compound heterozygote, with a single missense mutation in one allele, which resulted in Gly57Ser amino acid substitution, and a single C insertion at nucleotide position 899 in the second allele, which gave rise to a frame shift and premature termination codon. Case 2 was homozygous for the same C899 insertion found in case 1. PMID- 12649069 TI - Circadian rhythmicity of salivary leptin in healthy subjects. AB - In humans, circulating leptin correlates with body weight and fat mass. The main source of leptin is adipose tissue although placenta has been shown to produce leptin. More recently leptin and its receptor have been found in gastric mucosa and salivary gland. In the present study, we set to confirm the presence of leptin in human saliva and salivary gland, and if present, investigate whether salivary leptin exhibited circadian rhythmicity. Saliva and plasma were collected at 2 h intervals for 24 h in 12 healthy volunteers (6 females; 6 males). As previously described in human saliva and salivary gland, we have confirmed the presence of leptin at both mRNA and protein level. Leptin concentrations were significantly higher in plasma than saliva (p = 0.002), and a strong correlation between salivary and plasma leptin was demonstrated (r = 0.76; p < 0.001). Salivary leptin, like plasma leptin, showed circadian variations in both men and women, with a peak around 2400 h and a nadir at 1000 h. Both, mean 24-h salivary leptin and plasma leptin concentrations were significantly higher in women than men (p < 0.05; p < 0.01, respectively), but the ratios of salivary and plasma leptin concentrations were higher in men (p < 0.05), independent of other variables measured. In this study we have confirmed that leptin is synthesised and secreted by the human salivary gland, and have demonstrated for the first time that salivary leptin shows circadian variation. The precise role of salivary leptin needs to be elucidated. Our findings suggest that there may be differential regulation of leptin (salivary) between men and women. Finally, measurement of leptin in saliva is a simple, non-invasive and may be an acceptable alternative to plasma sampling. PMID- 12649070 TI - PAF- and bradykinin-induced hyperpermeability of rat venules is independent of actin-myosin contraction. AB - We tested the hypothesis that acutely induced hyperpermeability is dependent on actin-myosin contractility by using individually perfused mesentery venules of pentobarbital-anesthetized rats. Venule hydraulic conductivity (Lp) was measured to monitor hyperpermeability response to the platelet-activating factor (PAF) 1-O hexadecyl-2-acetyl-sn-glycero-3-phosphocholine or bradykinin. Perfusion with PAF (10 nM) induced a robust transient high Lp [24.3 +/- 1.7 x 10-7 cm/(s.cmH2O)] that peaked in 8.9 +/- 0.5 min and then returned toward control Lp [1.6 +/- 0.1 x 10-7 cm/(s.cmH2O)]. Reconstruction of venular segments with the use of transmission electron microscopy of serial sections confirmed that PAF induces paracellular inflammatory gaps. Specific inhibition of myosin light chain kinase (MLCK) with 1-10 microM 1-(5-iodonaphthalene-1-sulfonyl)-1H-hexahydro-1,4 diazepine hydrochloride (ML-7) failed to block the PAF Lp response or change the time-to-peak Lp. ML-7 reduced baseline Lp 50% at 40 min of pretreatment. ML-7 also increased the rate of recovery from PAF hyperpermeability measured as the decrease of half-time of recovery from 4.8 +/- 0.7 to 3.2 +/- 0.3 min. Inhibition of myosin ATPase with 5-20 mM 2,3-butanedione 2-monoxime also failed to alter the hyperpermeability response to PAF. Similar results were found using ML-7 to modulate responses. These experiments indicate that an actin-myosin contractile mechanism modulated by MLCK does not contribute significantly to the robust initial increase in permeability of rat venular microvessels exposed to two common inflammatory mediators. The results are consistent with paracellular gap formation by local release of endothelial-endothelial cell adhesion structures in the absence of contraction by the actin-myosin network. PMID- 12649071 TI - Remodeling of the skeletal muscle microcirculation increases resistance to perfusion in obese Zucker rats. AB - Whereas previous studies have demonstrated that the development of syndrome X in obese Zucker rats (OZR) is associated with impaired arteriolar reactivity to vasoactive stimuli, additional results from these studies indicate that the passive diameter of skeletal muscle arterioles is reduced in OZR versus lean Zucker rats (LZR). On the basis of these prior observations, the present study evaluated structural alterations to the skeletal muscle microcirculation as potential contributors to an elevated vascular resistance. Isolated skeletal muscle resistance arterioles exhibited a reduced passive diameter at all levels of intralumenal pressure and a left-shifted stress-strain curve in OZR versus LZR, indicative of structural remodeling of individual arterioles. Histological analyses using Griffonia simplicifolia I lectin-stained sections of skeletal muscle demonstrated reduced microvessel density (rarefaction) in OZR versus LZR, suggesting remodeling of entire microvascular networks. Finally, under maximally dilated conditions, constant flow-perfused skeletal muscle of OZR exhibited significant elevations in perfusion pressure versus LZR, indicative of an increased resistance to perfusion within the microcirculation. These data suggest that developing structural alterations to the skeletal muscle microcirculation in OZR result in elevated vascular resistance, which may, acting in concert with impaired arteriolar reactivity, contribute to blunted active hyperemic responses and compromised performance of in situ skeletal muscle with elevated metabolic demand. PMID- 12649073 TI - Attenuation of heart failure due to coronary stenosis by ACE inhibitor and angiotensin receptor blocker. AB - It is not known how the angiotensin-converting enzyme (ACE) inhibitor and angiotensin II receptor blocker (ARB) attenuate heart failure (HF) in viable ischemic hearts. To assess HF in a rat coronary stenosis (CS) model, we administered vehicle and quinapril or candesartan (or both) orally for 12 wk. Compared with the sham group, the vehicle group showed impaired myocardial perfusion, impaired coronary endothelial nitric oxide (NO) function in vitro, exhausted myocardial mitochondrial respiration, larger left ventricular (LV) dimensions and lower ejection fraction, lower LV rate of pressure development over time (dP/dt), lower slopes of LV end-systolic pressure-dimension relations (ESPDRs), and increased myocardial fibrosis. Treatment with quinapril or candesartan ameliorated these parameters without modifying the epicardial CS severity. Moreover, their combination maintained similar myocardial perfusion, despite a trend toward lower blood pressure, and showed distinctive neurohumoral modulation, normalized mitochondrial respiration, and increased ESPDR slopes. Thus improved myocardial blood flow and coronary flow reserve by quinapril or candesartan are the key to alleviate CS-induced HF, and their combination may have a therapeutic significance partly through ameliorated mitochondrial respiration and improved LV systolic function. PMID- 12649072 TI - Calcium dynamics in the failing heart: restoration by beta-adrenergic receptor blockade. AB - Changes in calcium (Ca2+) regulation contribute to loss of contractile function in dilated cardiomyopathy. Clinical treatment using beta-adrenergic receptor antagonists (beta-blockers) slows deterioration of cardiac function in end-stage heart failure patients; however, the effects of beta-blocker treatment on Ca2+ dynamics in the failing heart are unknown. To address this issue, tropomodulin overexpressing transgenic (TOT) mice, which suffer from dilated cardiomyopathy, were treated with a nonselective beta-receptor blocker (5 mg. kg-1. day-1 propranolol) for 2 wk. Ca2+ dynamics in isolated cardiomyocytes of TOT mice significantly improved after treatment compared with untreated TOT mice. Frequency-dependent diastolic and Ca2+ transient amplitudes were returned to normal in propranolol-treated TOT mice and but not in untreated TOT mice. Ca2+ kinetic measurements of time to peak and time decay of the caffeine-induced Ca2+ transient to 50% relaxation were also normalized. Immunoblot analysis of untreated TOT heart samples showed a 3.6-fold reduction of sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA), whereas Na+/Ca2+ exchanger (NCX) concentrations were increased 2.6-fold relative to nontransgenic samples. Propranolol treatment of TOT mice reversed the alterations in SERCA and NCX protein levels but not potassium channels. Although restoration of Ca2+ dynamics occurred within 2 wk of beta-blockade treatment, evidence of functional improvement in cardiac contractility assessed by echocardiography took 10 wk to materialize. These results demonstrate that beta-adrenergic blockade restores Ca2+ dynamics and normalizes expression of Ca2+-handling proteins, eventually leading to improved hemodynamic function in cardiomyopathic hearts. PMID- 12649074 TI - Expression of VEGF and angiopoietins-1 and -2 during ischemia-induced coronary angiogenesis. AB - The mechanisms underlying coronary capillary growth in response to ischemia are undefined. We hypothesized that the expression of vascular endothelial growth factor (VEGF) and angiopoietin (Ang)/Tie-2 were involved in capillary growth as an adaptation to ischemia. To test this hypothesis we measured capillary density, and the expressions of VEGF, Ang-1, Ang-2, and the Tie-2 receptor and its phosphorylation state during repetitive episodes of myocardial ischemia in chronically instrumented canines. Repetitive episodes of ischemia were induced by multiple (once/hour; 8/day), brief (2 min) occlusions of the left anterior descending coronary artery for 1, 7, 14, or 21 days. A sham group received the same instrumentation as the experimental groups but not the occlusion protocol. Collateral blood flow (microspheres) progressively increased from 9 +/- 3 to 83 +/- 10 ml. min-1. 100 g-1 on day 21. Capillary density increased at day 7 from 2378 +/- 53 (sham) to 2962 +/- 60/mm2, but it decreased to 2594 +/- 39/mm2 at day 21. Both VEGF and Ang-2 expression in myocardial interstitial fluid (Western analyses) peaked at day 3 of the repetitive occlusions but waned thereafter. In contrast the expression of Ang-1 remained relatively constant at all times in the occlusion groups. In shams, the expression of VEGF and Ang-2 was low and constant at all times. Tie-2 phosphorylation myocardial decreased decreased at day 7 but increased at 21 days of occlusions (P < 0.05). Our results indicate that capillary density was augmented by myocardial ischemia, but after development of collaterals and restoration of flow to the ischemic zone, capillary density returned to control levels. The change in capillary density paralleled with VEGF and Ang-2 expression but was inversely related to Tie-2 phosphorylation. We speculate the coronary angiogenesis is a coordinated event involving the expression of both VEGF and Ang-2 and that therapeutic angiogenic strategies may ultimately require treatment with more than a single factor. PMID- 12649076 TI - Caveolae-associated proteins in cardiomyocytes: caveolin-2 expression and interactions with caveolin-3. AB - Caveolin-3 the muscle-specific caveolin isoform, acts like the more ubiquitously expressed caveolin-1 to sculpt caveolae, specialized membrane microdomains that serve as platforms to organize signal transduction pathways. Caveolin-2 is a structurally related isoform that alone does not drive caveolae biogenesis; rather, caveolin-2 cooperates with caveolin-1 to form caveolae in nonmuscle cells. Although caveolin-2 might be expected to interact in an fashion analogous to that of caveolin-3, it generally has not been detected in cardiomyocytes. This study shows that caveolin-2 and caveolin-3 are detected at low levels in ventricular myocardium and increase dramatically with age or when neonatal cardiomyocytes are placed in culture. In contrast, flotillins (caveolin functional homologs) are expressed at relatively constant levels in these preparations. In neonatal cardiac cultures, caveolin-2 and -3 expression is not influenced by thyroid hormone (a postnatal regulator of other cardiac gene products). The further evidence that caveolin-2 coimmunoprecipitates with caveolin-3 and floats with caveolin-3 by isopycnic centrifugation in cardiomyocyte cultures suggests that caveolin-2 may play a role in caveolae biogenesis and influence cardiac muscle physiology. PMID- 12649075 TI - Glutamate release in midbrain periaqueductal gray by activation of skeletal muscle receptors and arterial baroreceptors. AB - We have previously reported that both skeletal muscle receptor and arterial baroreceptor afferent inputs activate neurons in the dorsolateral (DL) and lateral regions of the midbrain periaqueductal gray (PAG). In this study, we determined whether the excitatory amino acid glutamate (Glu) is released to mediate the increased activity in these regions. Static contraction of the triceps surae muscle for 4 min was evoked by electrical stimulation of the L7 and S1 ventral roots in cats. Activation of arterial baroreceptor was induced by intravenous injection of phenylephrine. The endogenous release of Glu from the PAG was recovered with the use of a microdialysis probe. Glu concentration was measured by the HPLC method. Muscle contraction increased mean arterial pressure (MAP) from 98 +/- 10 to 149 +/- 12 mmHg (P < 0.05) and increased Glu release in the DL and lateral regions of the middle PAG from 0.39 +/- 0.10 to 0.73 +/- 0.12 microM (87%, P < 0.05) in intact cats. After sinoaortic denervation and vagotomy were performed, contraction increased MAP from 95 +/- 12 to 158 +/- 15 mmHg, and Glu from 0.34 +/- 0.08 to 0.54 +/- 0.10 microM (59%, P < 0.05). The increases in arterial pressure and Glu were abolished by muscle paralysis. Phenylephrine increased MAP from 100 +/- 13 to 162 +/- 22 mmHg and increased Glu from 0.36 +/- 0.10 to 0.59 +/- 0.18 microM (64%, P < 0.05) in intact animals. Denervation abolished this Glu increase. Summation of the changes in Glu evoked by muscle receptor and arterial baroreceptor afferent inputs was greater than the increase in Glu produced when both reflexes were activated simultaneously in intact state (123% vs. 87%). These data demonstrate that activation of skeletal muscle receptors evokes release of Glu in the DL and lateral regions of the middle PAG, and convergence of afferent inputs from muscle receptors and arterial baroreceptors in these regions inhibits the release of Glu. These results suggest that the PAG is a neural integrating site for the interaction between the exercise pressor reflex and the arterial baroreceptor reflex. PMID- 12649077 TI - Exercise improves postischemic function in aging hearts. AB - Exercise improves cardioprotection against ischemia-reperfusion in young animals but has not been investigated in older animals, which represent the population most likely to suffer an ischemic event. Therefore, we sought to determine the effects of aging on exercise-induced cardioprotection. Young, middle-aged, and old (4, 12, and 21 mo old) male Fischer 344 rats ran 60 min at 70-75% of maximum oxygen consumption. Twenty-four hours postexercise, isolated perfused working hearts underwent 22.5 min of global ischemia and then 30 min of recovery (reperfusion). Compared with sedentary rats (n = 8-9 rats/group), recovery of function (cardiac output x systolic pressure) improved after exercise (n = 9 rats/group) by 40% at 4 mo, 78% at 12 mo, and 59% at 21 mo. Exercise increased inducible heat shock protein 70 expression 105% at 4 mo but only 27% at 12 mo and 24% at 21 mo. Catalase activity progressively increased with age (P < 0.05) and was increased by exercise at 4 mo (26%) and 21 mo (19%). Manganese superoxide dismutase activity was increased by exercise only at 21 mo (45%). No exercise related change in any antioxidant enzyme was observed at 12 mo. We conclude that exercise can enhance cardioprotection regardless of age, but the cardioprotective protein phenotype changes with age. PMID- 12649078 TI - Modulation of protein phosphatase 2a by adenosine A1 receptors in cardiomyocytes: role for p38 MAPK. AB - Adenosine A1 receptor activation causes protein phosphatase 2a (PP2a) activation in ventricular myocytes. This attenuates beta-adrenergic functional effects in the heart (Liu Q and Hofmann PA. Am J Physiol Heart Circ Physiol 283: H1314 H1321, 2002). The purpose of the present study was to identify the signaling pathway involved in the translocation/activation of PP2a by adenosine A1 receptors in ventricular myocytes. We found that N6-cyclopentyladenosine (CPA; an adenosine A1 receptor agonist)-induced PP2a translocation was blocked by p38 MAPK inhibition but not by JNK inhibition. CPA increased phosphorylation of p38 MAPK, and this effect was abolished by pertussis toxin and inhibitors of the cGMP pathway. Moreover, CPA-induced PP2a translocation was blocked by inhibition of the cGMP pathway. Guanylyl cyclase activation mimicked the effects of CPA and caused p38 MAPK phosphorylation and PP2a translocation. Finally, CPA-induced dephosphorylations of troponin I and phospholamban were blocked by pertussis toxin and attenuated by p38 MAPK inhibition. These results suggest that adenosine A1 receptor-mediated PP2a activation uses a pertussis toxin-sensitive Gi protein guanylyl cyclase-p38 MAPK pathway. This proposed, novel pathway may play a role in acute modulation of cardiac function. PMID- 12649079 TI - Effects of aging on capillary geometry and hemodynamics in rat spinotrapezius muscle. AB - The effects of aging on muscle microvascular structure and function may play a key role in performance deficits and impairment of O2 exchange within skeletal muscle of senescent individuals. To determine the effects of aging on capillary geometry, red blood cell (RBC) hemodynamics, and hematocrit in a muscle of mixed fiber type, spinotrapezius muscles from Fischer 344 x Brown Norway hybrid rats aged 6-8 mo [young (Y); body mass 421 +/- 10 g, n = 6] and 26-28 mo [old (O); 561 +/- 12 g, n = 6] were observed by high-resolution transmission light microscopy under resting conditions. The percentage of RBC-perfused capillaries (Y: 78 +/- 3%; O: 75 +/- 2%) and degree of tortuosity and branching (Y: 13 +/- 2%; O: 13 +/- 2%, additional capillary length) were not different in O vs. Y muscles. Lineal density of RBC-perfused capillaries in O was significantly reduced (Y: 30.7 +/- 1.8, O: 22.8 +/- 3.1 capillaries/mm; P < 0.05). However, RBC-perfused capillaries from O rats (n = 78) exhibited increased RBC velocity (VRBC) (Y: 219 +/- 12, O: 310 +/- 14 microm/s; P < 0.05) and RBC flux (FRBC) (Y: 27 +/- 2, O: 41 +/- 2 RBC/s; P < 0.05) vs. Y rats (n = 66). Thus O2 delivery per unit of muscle was not different between groups (Y: 894 +/- 111, O: 887 +/- 118 RBC. s-1. mm muscle-1). Capillary hematocrit was not different in Y vs. O rats (Y: 26 +/- 1%, O: 28 +/- 1%: P > 0.05). These data indicate that in resting spinotrapezius muscle, aging decreases the lineal density of RBC-perfused capillaries while increasing mean VRBC and FRBC within those capillaries. Whereas muscle conductive O2 delivery and capillary hematocrit were unchanged, elevated VRBC reduces capillary RBC transit time and may impair the diffusive transport of O2 from blood to myocyte particularly under exercise conditions. PMID- 12649080 TI - Do angiotensin-converting enzyme inhibitors directly stimulate the kinin B1 receptor? AB - It has been recently claimed that the human B1 receptors for kinins bind angiotensin-converting enzyme (ACE) inhibitors via a potential zinc-binding domain and are pharmacologically stimulated by these drugs. We verified whether ACE inhibitors stimulate B1 receptors in vitro. The isolated rabbit aorta or mouse stomach responded by negligible contractions to the application of captopril, enalaprilat, or zofenoprilat. The human isolated umbilical vein also failed to respond to enalaprilat. All of these preparations were responsive to the B1 receptor agonists des-Arg9-bradykinin (BK) or Lys-des-Arg9-BK. Furthermore, enalaprilat applied continuously had no significant interaction with the effects of Lys-des-Arg9-BK on the rabbit aorta. Enalaprilat failed to stimulate [3H]arachidonate release, translocate the receptors (confocal microscopy), or stimulate ERK1/2 phosphorylation (immunoblot) in HEK-293 cells stably expressing the rabbit B1 receptor conjugated to yellow fluorescent protein. The phospho-ERK1/2 content of arterial smooth muscle cells of human or rabbit origin was increased by treatment with Lys-des-Arg9-BK but not with enalaprilat. ACE inhibitors do not act as bona fide agonists of the kinin B1 receptors. PMID- 12649081 TI - Cardiac allograft vasculopathy and dysregulation of the NO synthase pathway. AB - Cardiac allograft vasculopathy is the most aggressive form of atherosclerosis in humans and is the leading cause of death after the first year of heart transplantation. Endothelial dysfunction is a major contributing factor to the acceleration of coronary vascular disease in these individuals. A reflection of this endothelial dysfunction is the severe impairment in endothelium-dependent vasodilation that occurs early after transplantation. The etiology of this allograft endothelial alteration is multifactorial and may include preexisting atherosclerosis of the graft vessels, reperfusion injury during transplantation, denervation, disruption of the lymphatic system, and acute and chronic immune injury, as well as traditional risk factors for coronary artery disease (hyperlipidemia, diabetes, hypertension, or hyperhomocysteinemia) and pathogens, such as cytomegalovirus. The alteration in endothelial function affects vasomotor tone of the coronary arteries. Evidence indicates that there may be an impairment of endothelial production and/or activity of NO. Because NO is a potent vasodilator, its deficiency would explain the abnormal vasomotor tone in these individuals. In addition, because NO inhibits key processes in vascular inflammation and atherosclerosis, its absence may contribute to the acceleration of transplant vascular disease. Recent studies from our group and others have shed light on the mechanisms of endothelial dysfunction and its importance in cardiac allograft vasculopathy. In addition, the alteration in endothelial function contributes to vascular inflammation and progression of the disease. PMID- 12649082 TI - Structure, function, and endothelium-derived hyperpolarizing factor in the caudal artery of the SHR and WKY rat. AB - OBJECTIVE: To quantify structural and functional characteristics of the caudal artery from spontaneously hypertensive (SHR) and normotensive Wistar Kyoto (WKY) rats with particular reference to endothelium-derived hyperpolarizing factor (EDHF). METHODS AND RESULTS: Ultrastructural studies showed that the number of myoendothelial gap junctions, smooth muscle cell (SMC) layers, and medial cross sectional area were significantly greater in SHR than WKY. Intracellular dye labeling demonstrated hyperplasia of SMCs in SHR. Analysis of nerve-mediated excitatory junction potentials recorded in SMCs at the adventitial and luminal borders demonstrated decreased radial coupling of SMCs in SHR. In both SHR and WKY, in the presence of NG-nitro-L-arginine methyl ester and indomethacin, acetylcholine-elicited EDHF was abolished by charybdotoxin and apamin, while iberiotoxin had no effect, implicating the involvement of small and intermediate, but not large, calcium-activated potassium channels. EDHF was abolished by Gap mimetic peptides, 18beta-glycyrrhetinic acid, and endothelial removal but not affected by the NO scavengers hydroxocobalamin and carboxy-PTIO. CONCLUSIONS: Significant differences in SMC morphology and homocellular and heterocellular coupling exist between the caudal artery of SHR and WKY rats. In the caudal artery of SHR, significantly greater heterocellular coupling compensates for other structural changes in the media to maintain a functional role for EDHF. PMID- 12649083 TI - White cell telomere length and risk of premature myocardial infarction. AB - OBJECTIVE: Biological age may be distinct from chronological age and contribute to the pathogenesis of age-related diseases. Mean telomeres lengths provide an assessment of biological age with shorter telomeres, indicating increased biological age. We investigated whether subjects with premature myocardial infarction (MI) had shorter leukocyte telomeres. METHODS AND RESULTS: Mean terminal restriction fragment (TRF) length, a measure of average telomere size, was compared in leukocyte DNA of 203 cases with a premature MI (<50 years) and 180 controls. Age- and sex-adjusted mean TRF length of cases was significantly shorter than that of controls (difference 299.7+/-69.3 base pairs, P<0.0001) and on average equivalent to controls 11.3 years older. The difference in mean TRF length between cases and controls was not accounted for by other coronary risk factors. Compared with subjects in the highest quartile for telomere length, the risk of myocardial infarction was increased between 2.8- and 3.2-fold (P<0.0001) in subjects with shorter than average telomeres. CONCLUSIONS: The findings support the concept that biological age may play a role in the etiology of coronary heart disease and have potentially important implications for our understanding of its genetic etiology, pathogenesis, and variable age of onset. PMID- 12649084 TI - E-selectin polymorphism associated with myocardial infarction causes enhanced leukocyte-endothelial interactions under flow conditions. AB - OBJECTIVE: Polymorphisms found in genes encoding adhesion molecules have been reported to be associated with atherosclerosis. We investigated the Ser128Arg polymorphism in the E-selectin gene in Japanese patients with myocardial infarction and its functional significance. METHODS AND RESULTS: Results from 135 patients with myocardial infarction and 327 control subjects revealed that the frequency of Arg128-positive was significantly higher in the patients than in controls (12.6% versus 6.7%; odds ratio, 2.0; 95% CI, 1.04 to 3.85), indicating that the Ser128Arg polymorphism was associated with myocardial infarction. We then generated a recombinant E-selectin adenovirus carrying a mutation (AdS128R E) and compared it with its wild-type counterpart by evaluating the adhesion characteristics of transduced human umbilical vein endothelial cells under flow. AdS128R-E-transduced human umbilical vein endothelial cells supported significantly more rolling and adhesion of neutrophils and mononuclear cells compared with human umbilical vein endothelial cells transduced with AdWT-E (P<0.001) and also exhibited significantly greater levels of phosphorylation of extracellular signal regulated kinase 1 and 2 and p38 mitogen-activated protein kinase, suggesting that an altered endothelial signaling pathway is associated with this polymorphism. CONCLUSIONS: Our results suggest that the E-selectin Ser128Arg polymorphism can functionally alter leukocyte-endothelial interactions as well as biochemical and biological consequences, which may account for the pathogenesis of myocardial infarction. PMID- 12649085 TI - Arterial remodeling and stiffness in patients with pseudoxanthoma elasticum. AB - OBJECTIVE: Proteoglycans organize the extracellular matrix, act as signaling molecules, and are involved in cell migration and proliferation. They may play an important role in arterial geometric and elastic properties. The aim of the present study was to determine large artery phenotype in patients with pseudoxanthoma elasticum (PXE), a genetic disease characterized by proteoglycan accumulation and fragmented elastic fibers in connective tissues. METHODS AND RESULTS: In 27 patients with PXE (25 females and 2 male) and 27 control subjects matched by age, sex, and blood pressure, we noninvasively determined the common carotid and radial artery diameter, intima-media thickness (IMT), and distensibility with high-definition echo-tracking systems and applanation tonometry. Patients with PXE had a significantly higher carotid IMT (611+/-106 versus 520+/-76 microm, P<0.001) independently of body surface area, age, and mean blood pressure. The increase in carotid IMT predominated in older patients with PXE at the time of examination. No significant difference in carotid elastic properties was observed between patients with PXE and control subjects. At the site of the radial artery, distensibility was significantly higher in patients with PXE than in control subjects (11.6+/-11.4 versus 5.9+/-3.4 kPa(-1) x 0.10( 3); P=0.02) and internal diameter was lower (1.66+/-0.51 versus 2.07+/-0.36 mm; P<0.01) without change in intima-media thickness and Young's elastic modulus. CONCLUSIONS: Phenotypic changes of superficial arteries in patients with PXE were represented by a thickening of the carotid artery and a reduced stiffness of the radial artery and predominated in older female patients. PMID- 12649086 TI - Expression of human scavenger receptor B1 on and in human platelets. AB - OBJECTIVE: The abundance of HDL particles correlates inversely with the incidence of coronary heart disease. The human scavenger receptor B1 (hSR-B1/CLA-1) is a receptor for HDL. Expression of hSR-B1/CLA-1 mRNA and protein in human platelets was determined using reverse transcriptase-polymerase chain reaction and Western blot, respectively. Presence of the protein on the surface of platelets was shown using flow cytometry. METHODS AND RESULTS: Immunochemical staining for hSR-B1/CLA 1 showed that it was expressed in megakaryocytes, the platelet precursors of human bone marrow. These findings prompted us to ask whether hSR-B1/CLA-1 was differentially expressed on platelets obtained from patients with atherosclerotic disease compared with those in control subjects. Our findings showed that abundance of hSR-B1/CLA-1 was significantly reduced on the surface of platelets from patients with atherosclerotic disease. The reduced levels of hSR-B1/CLA-1 were associated with increased cholesterol ester content in platelets from patients with atherosclerotic disease compared with control subjects. A negative correlation existed between hSR-B1/CLA-1 expression and platelet aggregation. In summary, our studies show that the HDL receptor hSR-B1/CLA-1 is expressed in platelets and their precursor, the megakaryocyte. The levels of hSR-B1/CLA-1 expression correlate inversely with cholesterol ester content and platelet aggregation. CONCLUSIONS: These findings suggest that determining the level of hSR-B1/CLA-1 expression on the platelets may be a useful clinical marker for atherosclerotic diseases. PMID- 12649087 TI - Soluble tissue factor induces coagulation on tumor endothelial cells in vivo if coadministered with low-dose lipopolysaccharides. AB - OBJECTIVE: This study was performed to evaluate the mechanisms leading to tumor vessel occlusion by tissue factor-based drugs, which are used in vascular targeting approaches for the treatment of malignant tumors. METHODS AND RESULTS: The effects of nontargeted soluble tissue factor were evaluated in vitro and in vivo. Tumor-bearing mice were treated with (1) the extracellular portion of tissue factor (soluble tissue factor), (2) low nontoxic doses of lipopolysaccharides, or (3) a combination thereof. The combination treatment showed the best effects and resulted in selective thrombosis of tumor vessels. On the basis of our data from subsequent in vitro analyses, including surface plasmon resonance measurements and endothelial cell based coagulation assays, we propose a model on how soluble tissue factor, although lacking its membrane anchor, can promote selective tumor vessel occlusion. CONCLUSIONS: To our knowledge, this is the first report to describe the molecular mechanisms of coagulation induction by untargeted soluble tissue factor in vivo. Combination treatments including soluble tissue factor might represent an alternative vascular targeting approach for the treatment of malignant tumors. PMID- 12649088 TI - Human plasma platelet-activating factor acetylhydrolase binds to all the murine lipoproteins, conferring protection against oxidative stress. AB - OBJECTIVE: Plasma platelet-activating factor (PAF) acetylhydrolase (AH) is an enzyme bound with lipoproteins that degrades not only PAF but also PAF-like oxidized phospholipids that are proposed to promote atherosclerosis. In this study, we investigated the distribution of PAF-AH protein among lipoprotein classes by using adenovirus-mediated gene transfer in mice, and we examined its effects on lipoprotein oxidation and foam cell formation of macrophages. METHODS AND RESULTS: Adenovirus-mediated overexpression of PAF-AH in mice resulted in a 76- to 140-fold increase in plasma PAF-AH activity. Contrary to the previous report, overexpressed human PAF-AH protein was bound to very low density lipoprotein, intermediate density lipoprotein, low density lipoprotein, and high density lipoprotein (HDL). All the lipoproteins with overexpressed human PAF-AH revealed more resistance against oxidative stress, which was associated with lower levels in autoantibody against oxidized low density lipoprotein in the plasma. In addition, HDL with human PAF-AH inhibited foam cell formation and facilitated cholesterol efflux in macrophages. CONCLUSIONS: These results suggest that human plasma PAF-AH exerts an antiatherogenic effect by binding to all the lipoproteins and thereby protecting them from oxidation, producing less proatherogenic lipoproteins and preserving HDL functions. PMID- 12649089 TI - Increased fecal bile acid excretion in transgenic mice with elevated expression of human phospholipid transfer protein. AB - OBJECTIVE: HDL plays a key role in protection against development of atherosclerosis by promoting reverse cholesterol transport from peripheral tissues to the liver for secretion into bile. Phospholipid transfer protein (PLTP) promotes the transfer of phospholipids between lipoproteins and modulates HDL size and composition, thereby having a crucial role in HDL metabolism. We investigated the effect of increased PLTP activity on removal of cholesterol from the body. METHODS AND RESULTS: On a chow diet, transgenic mice overexpressing human PLTP have a 15-fold increased plasma PLTP activity compared with wild-type mice (572.4+/-59.2 versus 38.6+/-3.6 micromol/mL per h). Plasma cholesterol, mainly present in HDL, is strongly decreased (-92%), caused by a rapid clearance from the circulation by the liver and leading to a 1.8-fold increase in hepatic cholesteryl esters. This results in a 2-fold increase in biliary bile acid secretion without changing the bile saturation index. Consequently, the transgenic mice show a 1.4-fold increase in the amount of excreted fecal bile acids compared with wild-type mice, whereas fecal neutral sterol excretion is unchanged. CONCLUSIONS: Our data show that elevation of PLTP activity results in rapid disposal of cholesterol from the body via increased conversion into bile acids and subsequent excretion. PMID- 12649090 TI - Effects of exercise on cardiovascular outcomes in monkeys with risk factors for coronary heart disease. AB - OBJECTIVE: Exercise reduces the risk for coronary heart disease. However, the mechanisms mediating the beneficial effects of exercise remain ambiguous. In particular, it is uncertain whether exercise inhibits the development of atherosclerosis, a major pathobiologic process underlying heart disease. METHODS AND RESULTS: To address this question, adult male monkeys were fed an atherogenic diet while assigned to one of four experimental conditions for 34 months: 1) runner/no group disruption, ie, "stable" (n=19); 2) runner plus frequent social group disruption, ie, "unstable" (n=19); 3) sedentary/stable (n=15); or 4) sedentary/unstable (n=18). Neither exposure to exercise nor social group disruption significantly affected the resulting coronary artery atherosclerosis extent or lumen areas (all ANOVA values, P>0.05). When compared with sedentary individuals, exercise animals had lower resting heart rates (119.0+/-3 vs 132.0+/ 3 bpm, P=0.002), greater echocardiographically measured left ventricular ejection fractions (77.2+/-0.01% vs 73.8+/-0.01%, P=0.02), greater quantitative angiographically measured dilation of coronary arteries to phenylephrine (2.6+/ 1% vs -3.7+/-1% change from baseline diameter, P=0.003), and a reduced cortisol response to an adrenocorticotropin challenge. These measures were not significantly affected by social condition. CONCLUSIONS: Thus, exercise improved some measures of cardiovascular health and reduced stress responsivity but did not inhibit progression of coronary artery atherosclerosis or promote positive artery remodeling. It is concluded that exercise may exert cardioprotective effects without influencing atherosclerosis extent. PMID- 12649092 TI - Inhibition of atherosclerosis in apoE-null mice by immunization with apoB-100 peptide sequences. AB - OBJECTIVE: LDL oxidation is believed to play an important role in the development of atherosclerosis, and oxidized LDL particles have been shown to become targets for the immune system. Immunization of animals with oxidized LDL results in reduction of atherosclerosis, suggesting an atheroprotective effect of this immune response. METHODS AND RESULTS: Using a polypeptide library covering the complete sequence of apoB-100, a large number of native and malondialdehyde modified peptide sequences in apoB-100 that are recognized by antibodies in human plasma were identified. We report here that immunization with apoB-100 peptide sequences, against which high levels of IgG and IgM antibodies are present in healthy human controls, reduce atherosclerosis in apoE-null mice by about 60%. Immunizations with these peptides were also found to increase the collagen content of subvalvular lesions. CONCLUSIONS: These studies have identified peptide sequences in apoB-100 that induce immune responses, which inhibits atherosclerosis. This suggests a way of developing an immunization therapy for coronary heart disease. PMID- 12649091 TI - Identification of immune responses against aldehyde-modified peptide sequences in apoB associated with cardiovascular disease. AB - OBJECTIVE: Atherosclerosis is associated with an immune response against oxidized LDL, which modulates the progression of the disease process. METHODS AND RESULTS: Using a library of polypeptides covering the complete sequence of apoB-100, the only major protein of LDL, we have identified over 100 different human antibodies reacting against aldehyde-modified apoB-100 sequences. IgM antibody titer levels decreased with age and were associated with the intima-media thickness of the carotid artery in subjects younger than 60 years. There were also inverse associations between IgM levels and oxidized LDL in plasma. In prospective clinical studies, antibody levels against several aldehyde-modified apoB-100 sites were associated with cardiovascular disease in this age group. Whether this immune response is adaptive (protective) or maladaptive (causal) in atherosclerosis requires further investigation. CONCLUSIONS: We have characterized a large number of epitopes within the apoB-100 component of oxidized LDL that provoke an immune response in humans. These findings will make it possible to study the role of immune responses against specific sites in oxidized LDL and to determine whether these immune responses influence the risk for future cardiac events. PMID- 12649093 TI - Can anything be done about oral mucositis? PMID- 12649094 TI - Chemoradiation therapy in locally advanced nasopharyngeal cancer: which kind of cooperation? PMID- 12649095 TI - Control of cancer-related anemia with erythropoietic agents: a review of evidence for improved quality of life and clinical outcomes. AB - BACKGROUND: Anemia occurs frequently in patients with cancer and is associated with impaired health-related quality of life (HRQOL). Treatment of anemia results in significant improvements in energy, activity and overall HRQOL, particularly among patients with mild-to-moderate anemia. Importantly, studies have indicated that anemia may have a negative impact on the success of radiotherapy, reducing survival and locoregional control. Recent preclinical and preliminary clinical data have also suggested that anemia may be associated with poorer outcomes following chemotherapy or surgery. MATERIALS AND METHODS: Data for review were identified and selected from searches of the literature published from January 1990 through to October 2002 using Medline, and searches of proceedings from key international oncology and hematology meetings. RESULTS: A wealth of data indicate that treatment of anemia improves HRQOL in patients with cancer. Prospective studies exploring survival and/or treatment outcomes in anemic cancer patients are currently in their early stages, preventing any firm conclusions from being drawn, although they do indicate a benefit in treating anemia. CONCLUSIONS: Recent studies support the use of erythropoietic agents in anemic cancer patients as a means of raising their hemoglobin levels and consequently improving their HRQOL. Randomized, controlled trials are needed to determine whether treating anemia with erythropoietic agents will improve other outcomes following therapy. PMID- 12649096 TI - An overview of the current clinical use of the anti-CD20 monoclonal antibody rituximab. AB - The chimeric anti-CD20 monoclonal antibody rituximab has become part of the standard therapy for patients with non-Hodgkin's lymphoma (NHL). To date, more than 300 000 patients have been treated with rituximab worldwide, including patients with indolent and aggressive NHL, Hodgkin's disease and other B-cell malignancies. Combination of rituximab with cytotoxic agents or cytokines has been explored in a number of different studies. Rituximab is now also approved for patients with diffuse large B-cell lymphoma when combined with standard CHOP chemotherapy (cyclophosphamide, doxorubicin, vincristine and prednisone). The monoclonal antibody is generally well tolerated. Most adverse events are infusion associated, including chills, fever and rigor related to the release of cytokines. PMID- 12649097 TI - Oxaliplatin plus high-dose leucovorin and 5-fluorouracil in pretreated advanced breast cancer: a phase II study. AB - BACKGROUND: The purpose of this study was to evaluate the efficacy and toxicity of oxaliplatin plus 5-fluorouracil (5-FU) and leucovorin (LV) in patients with metastatic breast cancer (MBC) previously treated with anthracyclines and taxanes. PATIENTS AND METHODS: Fifty anthracycline- and taxane-pretreated MBC patients were treated with oxaliplatin 85 mg/m(2) as a 2-h infusion on day 1, LV 200 mg/m(2)/day as a 2-h infusion followed by bolus 5-FU 400 mg/m(2)/day and a 22 h infusion of 5-FU 600 mg/m(2)/day for 2 consecutive days. Treatment was repeated every 3 weeks. Patients were evaluated for response every two cycles. RESULTS: The median age was 51 years (range 34-75). Twenty patients (40%) had received three or more chemotherapeutic regimens, 64% had three or four metastatic sites and 78% had visceral metastases. All patients had prior exposure to anthracyclines and taxanes. Based on an intention-to-treat analysis, one patient (2%) achieved a complete response and 16 (32%) a partial response, for a 34% overall response rate. Twenty-one patients (42%) had stable disease and 12 (24%) progressive disease. The median time to tumor progression was 5.3 months (range 0.5-12.8) and the median overall survival was 12.3 months (range 0.5-19.2). Toxicity was mild to moderate. Grade 3/4 neutropenia and thrombocytopenia occurred in 32% and 18%, respectively. Febrile neutropenia was experienced by three patients (6%), who were successfully treated. Grade 3/4 neurotoxicity was reported in 14% of the patients and gradually declined after treatment discontinuation. Cycle delays were reported in 28% of patients and dose reductions in 26%. Alopecia, nausea-vomiting, diarrhea and mucositis were not significant. There were no treatment-related deaths. CONCLUSION: The combination of oxaliplatin plus 5-FU/LV seems to be an active regimen in patients with MBC and prior exposure to anthracyclines and taxanes with a good safety profile. The incidence of severe toxicity was quite low and the compliance of patients to the treatment was satisfactory. The results obtained with this regimen could be considered encouraging in this heavily pretreated group of breast cancer patients with a high incidence of visceral metastases. PMID- 12649098 TI - A phase II study of ZD0473 given as a short infusion every 3 weeks to patients with advanced or metastatic breast cancer: a National Cancer Institute of Canada Clinical Trials Group trial, IND 129. AB - BACKGROUND: ZD0473 is a new generation platinum compound with activity against a wide range of human tumor cell lines and xenografts, including carboplatin- and cisplatin-resistant lines. A phase II study of ZD0473 in advanced breast cancer was initiated by the National Cancer Institute of Canada Clinical Trials Group. PATIENTS AND METHODS: Women with metastatic breast cancer, measurable disease, an Eastern Cooperative Oncology Group performance status of up to two, and a maximum of one prior cytotoxic agent for recurrent disease were enrolled and treated at 120 mg/m(2) every 3 weeks. After 13 patients were enrolled, the dose was increased to 150 mg/m(2) on the basis of emergent data from studies ongoing at the time. RESULTS: Thirty-three women were evaluable for toxicity and 26 patients for response. Toxicity was mainly hematological with grade 3/4 thrombocytopenia in 12 of 20 patients (60%) treated at 150 mg/m(2) and grade 3 thrombocytopenia in three of 13 patients (23%) at 120 mg/m(2). Grade 3/4 neutropenia occurred in 15 patients (75%) at 150 mg/m(2) and two patients (10%) at 120 mg/m(2). Non hematological toxicities were generally mild or moderate. There was one partial response seen for a response rate of 3.8% (95% confidence interval 0.1% to 19.5%) and stable disease in 15 patients. CONCLUSION: ZD0473 has minor activity as a single agent in metastatic breast cancer. Combinations with other drugs including docetaxel are ongoing and may be of interest. PMID- 12649099 TI - Predictive testing for BRCA1 and 2 mutations: a male contribution. AB - BACKGROUND: Management strategies for women carrying BRCA1 and 2 mutations are becoming clearer and predictive testing for a known family mutation is commonly undertaken. Implications for men are not as clear and they participate less frequently. PATIENTS AND METHODS: Twenty-six men from 10 extended families underwent predictive testing. Their motivation, reaction and outcome were studied. Subjects had appropriate pre- and post-test counselling. Informed consent was obtained before predictive testing for known deleterious mutations. DNA analysis followed standard procedures. RESULTS: Eighteen tested positive and eight negative. Four had adverse psychological reactions and three reneged on their commitments to impart results. The spouse of another man had an adverse psychological reaction to the disclosure of his positive result. Two, already suffering from prostate cancer, were phenocopies and paternal lineage transmission was unexpectedly determined in another. Risk was removed from 33 offspring and confirmed for 56. CONCLUSIONS: Complex themes associated with genetic testing are confirmed and the spectrum extended. Men appear to understand the importance of participating in this process. Methods of avoiding adverse reactions merit further study along with other aspects of the process. PMID- 12649100 TI - Low incidence of secondary myelodysplasia and acute myeloid leukemia after high dose chemotherapy as adjuvant therapy for breast cancer patients: a study by the Solid Tumors Working Party of the European Group for Blood and Marrow Transplantation. AB - BACKGROUND: To determine the incidence of secondary myelodysplasia (sMDS) or acute myeloid leukemia (AML) in node-positive breast cancer patients who received high-dose chemotherapy (HDCT) followed by autologous stem-cell support as adjuvant therapy. PATIENTS AND METHODS: The incidence of sMDS/AML was retrospectively assessed in 364 node-positive breast cancer patients who received HDCT followed by autologous stem-cell support as adjuvant therapy between November 1989 and December 1997 and were reported to the European Group for Blood and Marrow Transplantation registry. RESULTS: The median age of the patients was 45 years (range 22-62 years). Two hundred and ninety-one patients received peripheral blood stem cells and 55 patients received autologous bone marrow as stem-cell support. The most frequently used conditioning regimen was the STAMP-V regimen (32%), followed by melphalan-thiotepa (22%) and melphalan-mitoxantrone cyclophosphamide (21%). The 5-year probability of overall survival is 71% (95% CI 65% to 77%). After a median follow-up of 48 months (range 1-108 months) only one case of AML was observed, resulting in a crude incidence of 0.27%. This case of AML was observed 18 months after HDCT consisting of three cycles of epirubicin and cyclophosphamide with a cumulative dose of epirubicin 960 mg and cyclophosphamide 19 g. The French-American-British type of AML was M4, and the cytogenetic analysis showed a translocation t(9;11)(p22;q23). After complete remission following high-dose cytarabine and idarubicin the patient relapsed and died. CONCLUSIONS: In contrast to patients with malignant lymphoma there seems to be no increased risk of sMDS/AML after HDCT in breast cancer. Continued monitoring is required to confirm this low incidence after a longer follow-up period. PMID- 12649101 TI - Prophylaxis with GM-CSF mouthwashes does not reduce frequency and duration of severe oral mucositis in patients with solid tumors undergoing high-dose chemotherapy with autologous peripheral blood stem cell transplantation rescue: a double blind, randomized, placebo-controlled study. AB - BACKGROUND: The aim of this study was to evaluate the effectiveness of granulocyte-macrophage colony-stimulating factor (GM-CSF) mouthwashes in the prevention of severe mucositis induced by high doses of chemotherapy. PATIENTS AND METHODS: Ninety consecutive patients affected by solid tumors and undergoing high-dose chemotherapy with autologous peripheral blood stem cell transplantation rescue were randomized to receive placebo versus GM-CSF mouthwash 150 micro g/day. Patients were stratified on the basis of the conditioning treatment and the consequent different risk of severe oral mucositis. Treatment was administered from the day after the end of chemotherapy until the resolution of stomatitis and/or neutrophil recovery. RESULTS: The statistical analyses were intention-to-treat and involved all patients who entered the study. The severity of stomatitis was evaluated daily by the physicians according to National Cancer Institute Common Toxicity Criteria. Both study and control groups were compared with respect to the frequency [30% versus 36%, chi(2) exact test, not significant (NS)] and mean duration (4.8 +/- 4.7 versus 4.4 +/- 2.7 days, t-test, NS) of severe stomatitis (grade > or =3). Oral pain was evaluated daily by patients themselves by means of a 10 cm analog visual scale: the mean (+/- standard error of the mean) maximum mucositis scores were 4.8 +/- 3.5 versus 4.2 +/- 3.5 cm (t test, NS). Furthermore, 15/46 patients in the study group (33%) and 19/44 patients in the control group experienced pain requiring opioids (chi(2) exact test, NS). CONCLUSION: We did not find any evidence to indicate that prophylaxis with GM-CSF mouthwash can help to reduce the severity of mucositis in the setting of the patients we studied. PMID- 12649102 TI - Induction chemotherapy followed by concomitant chemoradiotherapy in the treatment of locoregionally advanced nasopharyngeal cancer. AB - BACKGROUND: Since 1990, we have treated patients with advanced nasopharyngeal cancer with induction chemotherapy and concomitant chemoradiotherapy. We herein report the results of our experience. PATIENTS AND METHODS: From 1990 to 1999, 27 patients with locoregionally advanced nasopharyngeal cancer were treated with induction chemotherapy followed by concomitant chemoradiotherapy. Using the American Joint Committee on Cancer's 1992 stage classification, all patients were stage III (11%) or IV (89%). By histology, 63% were poorly differentiated carcinoma and 37% squamous cell carcinoma. The median age was 42 years. Three cycles of induction chemotherapy consisting of cisplatin, 5-fluorouracil, leucovorin and interferon-alpha2b were administered, followed by concomitant chemoradiotherapy consisting of seven cycles of 5-fluorouracil, hydroxyurea and once-daily radiotherapy (FHX) on a week-on week-off schedule. The median radiotherapy dose was 70 Gy. RESULTS: Clinical response to induction chemotherapy was 100%, 54.2% complete response (CR) and 45.8% partial response. Clinical and/or pathological (37% of all patients had post-treatment biopsy with or without neck dissection) CR after FHX was 100%. At a median follow-up of 52 months, three failures were observed. Two patients have died of disease, one of local failure and one of distant metastases. One patient is alive with an isolated rib metastasis. At 5 years, actuarial locoregional control is 93% and actuarial distant control 92%. The overall survival at 3 and 5 years is 88% and 77%, respectively. Four patients died of unrelated illnesses and had no evidence of disease with respect to their nasopharyngeal cancer. The progression-free survival at 3 and 5 years is 92% and 86%, respectively. Thirty-three per cent of patients required a reduction in the chemotherapy dose due to acute toxicity. Chronic toxicity was not observed, with all patients able to eat orally without dietary restrictions. CONCLUSIONS: Treatment of locoregionally advanced nasopharyngeal cancer with induction chemotherapy followed by concomitant chemoradiotherapy resulted in excellent overall survival with acceptable toxicity. These results are encouraging and warrant further investigation of intensified approaches. PMID- 12649103 TI - Raltitrexed plus radiotherapy for the treatment of unresectable/recurrent rectal cancer: a phase I study. AB - BACKGROUND: Current consensus is that a combination of radiotherapy and chemotherapy may provide the optimal treatment for patients with unresectable rectal cancer. Raltitrexed has proven efficacy in the treatment of advanced colorectal cancer and has an acceptable toxicity profile. The aim of this phase I study was to determine the recommended dose of raltitrexed in combination with radiotherapy in patients with unresectable/recurrent rectal cancer. PATIENTS AND METHODS: Patients were treated with radiotherapy (25 fractions at 2.0 Gy per fraction) five times per week for 5 weeks. Raltitrexed was administered on days 1 and 22 at 2.0, 2.6 and 3.0 mg/m(2). RESULTS: A total of 20 patients were entered into the study. Dose-limiting toxicities were recorded in three of 20 patients following the first dose of raltitrexed; one patient at 2.6 mg/m(2) (grade 3 diarrhoea, grade 3 neutropenia and grade 2 pyrexia) and two patients at 3.0 mg/m(2) (one grade 3 neutropenia and one grade 4 diarrhoea). The most common non haematological and haematological treatment-related adverse events were diarrhoea (11 of 20, two grade 3, one grade 4) and leukopenia (eight of 20, one grade 3, one grade 4), respectively. CONCLUSIONS: The recommended dose of raltitrexed in combination with radiotherapy for future studies is 2.6 mg/m(2). PMID- 12649104 TI - Activity of raltitrexed and gemcitabine in advanced pancreatic cancer. AB - BACKGROUND: Gemcitabine has evolved as standard therapy in advanced pancreatic cancer since the demonstration of a significant clinical benefit. Phase II trials have shown that gemcitabine can be successfully combined with thymidylate synthase (TS) inhibitors such as continuous-infusion 5-fluorouracil (5-FU). However, continuous-infusion 5-FU is inconvenient because of the need for a central venous access. The aim of this study was to assess the efficacy and safety of gemcitabine in combination with raltitrexed (Tomudex), a novel and selective TS inhibitor that has the advantage of a 3-weekly treatment interval and manageable toxicity. PATIENTS AND METHODS: Chemotherapy-naive patients with measurable advanced pancreatic cancer were treated with raltitrexed 3 mg/m(2) as a 15-min infusion on day 1 and gemcitabine 1000 mg/m(2) on days 1 and 8, every 21 days. RESULTS: Twenty-five eligible patients (17 male, eight female) with metastatic (21 patients) or locally advanced (four patients) disease entered the study. The median number of courses per patient was four (range 1-14). One patient was not evaluable for response. There were three partial remissions [12%; 95% confidence interval (CI) 2.6% to 31.2%] and nine stable disease situations (36%; 95% CI 18.0% to 57.5%), while the tumours of 12 patients (48%; 95% CI 27.8% to 68.7%) showed progressive disease after three treatment cycles. WHO grade 3/4 toxicity was rare and symptomatic in only one patient, who experienced grade 4 diarrhoea and grade 3 nausea and vomiting. Symptomatic benefit was seen in 12 patients. Median survival was 185 days (95% CI 129-241) with six patients still alive. CONCLUSIONS: The efficacy of raltitrexed plus gemcitabine is limited, but compares well with other chemotherapy treatment options in advanced pancreatic cancer. However, this combination is convenient and symptomatic toxicity is rare. Thus, raltitrexed and gemcitabine should be investigated further in combination with drugs interfering with specific molecular targets. PMID- 12649105 TI - Gemcitabine and oxaliplatin for metastatic pancreatic adenocarcinoma: a North Central Cancer Treatment Group phase II study. AB - BACKGROUND: This study was performed to determine the efficacy of gemcitabine and oxaliplatin in patients with advanced or metastatic pancreatic adenocarcinoma (ACA). PATIENTS AND METHODS: Pancreatic ACA patients with previously untreated advanced or metastatic disease were enrolled in a phase II study of gemcitabine and oxaliplatin. Oxaliplatin was given i.v. on day 1 and gemcitabine i.v. on days 1 and 8 of a 3-week cycle. The primary end point of the trial was 6-month survival. Secondary end points included response rate, overall survival, median time to progression and toxicity. RESULTS: A total of 47 patients were enrolled, 46 of whom were evaluable. Of those patients assessed for the primary end point 50% lived for > or =6 months. The median time to progression was 4.53 months. Five confirmed responses were seen with a median duration of response of 2.7 months. Overall, the treatment was well tolerated. However, one patient died as a result of treatment-related hemolytic uremic syndrome. CONCLUSIONS: Gemcitabine and oxaliplatin, at doses of 1000 mg/m(2) and 100 mg/m(2), respectively, showed moderate activity in patients with pancreatic ACA. Based on the results of this study further evaluation of this combination is warranted. PMID- 12649107 TI - Prognostic factors for patients with advanced stage serous borderline tumours of the ovary. AB - BACKGROUND: The aim of this study was to determine the prognostic factors for patients with advanced stage, low malignant potential ovarian tumour (LMPOT). PATIENTS AND METHODS: A retrospective review of 80 patients with serous LMPOT and peritoneal implants treated at or referred to our institution was carried out. RESULTS: Sixty-five patients had non-invasive implants. Fifteen patients had invasive implants. Twenty-nine patients had stage II and 51 patients had stage III disease. Three patients died of evolutive invasive disease and four of complications of treatment. The only prognostic factor of progression to 'evolutive invasive disease' is the pathologic subtype of peritoneal implants. The 5-year rates of evolutive invasive disease in patients with non-invasive implants and invasive implants were 2% and 31%, respectively (P <0.002). CONCLUSIONS: In this series, the only prognostic factor for patients with advanced stage borderline tumour is the type of peritoneal implant. More patients died of the treatment's complications than of the disease itself. The patients' prognosis with non-invasive implants seems to be excellent, and conservative management could be discussed in younger patients. PMID- 12649106 TI - Staging of digestive endocrine tumours using helical computed tomography and somatostatin receptor scintigraphy. AB - BACKGROUND: In patients with digestive endocrine tumours, complete pre-operative staging is essential in planning proper management and evaluating treatment efficacy. To date, somatostatin receptor scintigraphy (SRS) is considered the 'gold standard' imaging procedure, and very few data are available concerning the use of helical computed tomography (hCT). This study aimed to determine the diagnostic accuracy and the ability to modify the surgical management of hCT, alone or combined with SRS. PATIENTS AND METHODS: Sixty patients were staged before surgery by hCT, SRS and tumour markers, and included in group 1 if suitable for radical surgery, otherwise in group 2. All patients underwent laparotomy followed by subsequent re-staging. RESULTS: SRS sensitivity was 77%, 48% and 67% for primary, lymph-node and liver lesions, respectively. hCT sensitivity was 94%, 69% and 94% for primary, lymph-node and liver lesions, respectively (P = 0.02 versus SRS, for liver lesions). During pre-operative evaluation, hCT correctly staged 92% and SRS 75% of patients (P = 0.02). hCT provided additional information in 17% of patients. CONCLUSIONS: Since hCT has been shown to be extremely accurate, providing essential information for the planning of surgical treatment compared with that of SRS, both techniques should be used in the pre-operative work-up of digestive endocrine tumours. PMID- 12649108 TI - Second-line chemotherapy with temozolomide in recurrent oligodendroglioma after PCV (procarbazine, lomustine and vincristine) chemotherapy: EORTC Brain Tumor Group phase II study 26972. AB - BACKGROUND: Oligodendroglial tumors are chemosensitive, with two-thirds of patients responding to PCV combination chemotherapy with procarbazine, lomustine (CCNU) and vincristine. Temozolomide (TMZ), a new alkylating and methylating agent has shown high response rates in recurrent anaplastic astrocytoma. We investigated this drug in recurrent oligodendroglial tumors (OD) and mixed oligoastrocytomas (OA) after prior PCV chemotherapy and radiation therapy. PATIENTS AND METHODS: In a prospective non-randomized multicenter phase II trial patients were treated with TMZ 150 mg/m(2) on days 1-5 in cycles of 28 days for 12 cycles. Eligible patients had a recurrence after prior PCV chemotherapy, with measurable and enhancing disease as shown by magnetic resonance imaging. Pathology and all responses were centrally reviewed. RESULTS: Thirty-two eligible patients were included. In four patients the pathology review did not confirm the presence of an OD or OA. Twelve of 24 patients [50%, 95% confidence interval (CI) 29% to 71%] evaluable for response to first-line PCV chemotherapy had responded to PCV. Temozolomide was in general well tolerated; the most frequent side effects were hematological. One patient discontinued treatment due to toxicity. In seven of 28 patients (25%, 95% CI 11% to 45%) with histologically confirmed OD an objective response to TMZ was observed. Median time to progression for responding patients was 8.0 months. After 6 and 12 months from the start of treatment, 29% and 11% of patients, respectively, were still free from progression. CONCLUSIONS: TMZ may be regarded as the preferred second-line treatment in OD after failure of PCV chemotherapy. Further studies on TMZ in OD are indicated. PMID- 12649109 TI - Multicentre phase II study and pharmacokinetic analysis of irinotecan in chemotherapy-naive patients with glioblastoma. AB - BACKGROUND: To assess the antitumour activity and safety profile of irinotecan and its pharmacokinetic interactions with anticonvulsants in patients with glioblastoma multiforme. PATIENTS AND METHODS: This multicentre phase II and pharmacokinetic study investigated the effects of irinotecan 350 mg/m(2) given as a 90-min infusion every 3 weeks either prior to (group A) or after relapse following radiotherapy (group B) in chemotherapy-naive patients with glioblastoma. Preferred concomitant medication for seizure prevention was valproic acid. Pharmacokinetic analysis of irinotecan and its main metabolites (SN-38, SN-38-G, APC and NPC) was performed during cycle 1. An independent panel of experts reviewed the activity data. RESULTS: Fifty-two patients (25 patients in group A and 27 patients in group B) received a total of 191 cycles of irinotecan. Forty-six patients (22 patients in group A and 24 patients in group B) were evaluable and externally reviewed for activity. According to external review, one partial response (group B), seven minor responses (three in group A and four in group B), 12 disease stabilisations (seven in group A and five in group B) were observed. This resulted in an overall response rate of only 2.2% (95% confidence interval 0.2% to 6.5%). The median time to tumour progression was 9 weeks in group A and 14.4 weeks in group B. Six-month progression-free survival rates were 26% in group A and 43% in group B. Grade 3-4 toxicities (percentage of patients in groups A and B) consisted of neutropenia (12.5% and 25.9%), diarrhoea (8.3% and 7.4%), asthenia (12.5% and 7.4%) and vomiting (0% and 7.4%). The clearance of irinotecan was 12.4 and 14.4 l/h/m(2) in two patients who received no anticonvulsant. In patients receiving valproic acid, the clearance of irinotecan was 17.2 +/- 4.4 l/h/m(2). CONCLUSIONS: Irinotecan given at the dose of 350 mg/m(2) every 3 weeks has limited clinical activity as a single agent in patients with newly diagnosed and recurrent glioblastoma after radiotherapy. The toxicity profile and plasma disposition of irinotecan and SN-38 were not strongly influenced by anticonvulsant valproic acid therapy. Although the response rate of irinotecan as a single agent was limited, it remains an attractive drug for combination studies in patients with glioblastoma. PMID- 12649110 TI - Clinical and pharmacokinetic phase II study of fotemustine in refractory and relapsing multiple myeloma patients. AB - BACKGROUND: Patients with relapsing or refractory multiple myeloma have poor prognosis. Few compounds are active in these patients and response duration remains short. We report the results of an open phase II trial evaluating the efficacy and safety of fotemustine monotherapy. PATIENTS AND METHODS: Twenty-one patients with relapsing (17) or refractory (four) multiple myeloma received fotemustine 100 mg/m(2) on an outpatient basis on days 1 and 8 of the induction cycle, followed after a 6-week rest period by fotemustine 100 mg/m(2) every 3 weeks until progression or unacceptable toxicity. Fotemustine pharmacokinetics during the first day of induction was compared between patients with normal or abnormal renal function. RESULTS: Five of 20 eligible patients had an objective response giving an intention-to-treat response rate of 25% [95% confidence interval (CI) 6% to 44%] and a 35.7% response rate (95% CI 11% to 61%) in the 14 patients having received at least four injections of fotemustine. The median time to objective response was 8.9 months. The median times to progression and survival were 13.8 and 23.1 months, respectively, with a 2-year survival rate of 49%. The main toxicity was myelosuppression with grade 3-4 neutropenia and thrombocytopenia in 66% and 71% of patients, respectively. There was one toxic death by sepsis after induction. The pharmacokinetic parameters in renal-impaired patients were not significantly different from those in patients with normal renal function with a similar incidence of grade 3-4 toxicity in both groups. CONCLUSIONS: Fotemustine as a single agent has definite activity in patients with relapsing or refractory multiple myeloma, with acceptable toxicity and can be administered at conventional doses in patients with mild or moderate renal impairment. PMID- 12649111 TI - Primary follicular lymphoma of the gastrointestinal tract: a study of 25 cases and a literature review. AB - BACKGROUND: To describe better the clinical, biological, endoscopic and pathological presentations, as well as the outcome, of primary follicular lymphoma (FL) of the gastrointestinal (GI) tract. PATIENTS AND METHODS: From November 1983 to February 2001, 25 eligible patients with primary FL of the GI tract were retrieved from several French Departments of Pathology departments based on histological diagnosis and immunophenotype. Median age was 56 years (range 44-71) with a sex ratio female/male of 2 (17/8). RESULTS: Abdominal pain was the main presenting symptom followed by intestinal obstruction. The small intestine was the most common site of involvement. Lesions were unifocal in the majority of patients (15/25). A pattern similar to lymphomatous polyposis was observed in 50% (7/14) of patients. Twelve patients had stage I, 10 patients stage II and three patients stage IV disease, and there was minimal extra intestinal involvement. Lymphoma tissues were composed of neoplastic follicles, most of which were grade 1 according to the World Health Organization (WHO) classification. The immunophenotype of the lymphoma cells was CD20+, CD10+, bcl2+ and CD5-. In tissue samples, IgH/bcl2 rearrangement at the MBR locus was present in 11 of 14 patients tested. Seven patients did not receive any treatment; four of them progressed after a median follow-up of 37.5 months. Treatment was otherwise heterogeneous, and complete remission was obtained in 15 patients which lasted for a median of 31 months. Relapses were either in the GI tract (n = 3) or outside the GI tract (n = 3). After a median follow-up of 34 months (range 5 203), 22 patients were still alive (complete remission, 11; partial remission, three; stable disease, six; progressive disease, two). CONCLUSIONS: Primary FL of the GI tract is a predominantly female lymphoma that most frequently involves the small intestine. Since the endoscopic and clinical presentation may not be different from lymphomatous polyposis, which is often associated with mantle cell origin of tumor cells, it is mandatory to perform an immunohistological and, if possible, a molecular analysis of GI lymphoma. The course of the disease is indolent and does not differ from nodal FL. Thus, therapy may not be required unless significant clinical symptoms are present or until disease progression. PMID- 12649112 TI - Phase I and pharmacokinetic study of the new vinca alkaloid vinflunine administered as a 10-min infusion every 3 weeks in patients with advanced solid tumours. AB - BACKGROUND: Vinflunine is a novel vinca alkaloid obtained by semi-synthesis using super-acidic chemistry to selectively introduce two fluorine atoms at the 20' position of vinorelbine. In human tumour xenografts, vinflunine showed definite antitumour activity in seven out of 11 tumours tested compared with three out of 11 for vinorelbine. PATIENTS AND METHODS: In this phase I study, vinflunine was administered to 31 patients with advanced malignancies as a 10-min i.v. infusion every 3 weeks according to an escalating schedule of doses between 30 and 400 mg/m(2). RESULTS: Pharmacokinetic parameters and toxicities were assessed and, at 400 mg/m(2), three out of five patients experienced dose-limiting toxicity. At the maximum tolerated dose (MTD), i.e. 400 mg/m(2), the toxicity profile of vinflunine consisted mainly of mucositis, constipation and neutropenia of short duration. Vinflunine area under the curve increased as a proportion of the administered dose whereas no saturation of elimination was observed. CONCLUSION: The MTD of vinflunine was achieved at 400 mg/m(2) every 3 weeks. According to protocol rules, the recommended dose was established at 350 mg/m(2). A preliminary assessment of first patients included in early phase II trials led to reduction of the recommended dose to 320 mg/m(2) every 3 weeks for further development of vinflunine. Three partial responses (two in breast carcinoma, one in renal cell carcinoma) suggest that activity is likely to be seen in less heavily pretreated patient populations. PMID- 12649113 TI - Phase I study of carboplatin, doxorubicin and weekly paclitaxel in patients with advanced ovarian carcinoma. AB - BACKGROUND: Doxorubicin is an active compound in epithelial ovarian cancer (EOC), but adding it to carboplatin-paclitaxel causes toxicity. Toxicity can be reduced by weekly administration. We examined the tolerability of weekly paclitaxel in combination with carboplatin and doxorubicin. PATIENTS AND METHODS: Chemotherapy naive patients with EOC were treated with doxorubicin (50 mg/m(2) day 1), carboplatin (AUC 6 day 1) and paclitaxel (days 1, 8, 15, 21), 28-day cycle. Three patients were treated at each paclitaxel dose level, starting at 60, 75 and 90 mg/m(2)/week. If more than two patients in a cohort experienced dose-limiting toxicity (DLT) three more patients were treated at the dose level below. RESULTS: Twelve patients with advanced EOC received a median of six cycles (range 2-6) of the three-drug combination. DLT occurred at dose level 3: prolonged grade 4 febrile neutropenia, 1 patient; grade 3 peripheral neuropathy, 1 patient. All six patients treated at dose level 2 experienced short-lived grade 4 neutropenia, which led to dose modifications resulting in an actual delivered dose of paclitaxel of 64 mg/m(2)/week. Eight out of 12 patients had measurable disease on CT scan: four obtained a partial remission; three had stable disease. CONCLUSIONS: The combination of carboplatin, doxorubicin and paclitaxel in patients with EOC is active and its main toxicity is myelosuppression. Dose intensity of paclitaxel can be maintained in a three-drug combination through weekly administration (65 mg/m(2)). PMID- 12649114 TI - Pharmacokinetics of low-dose carboplatin and applicability of a method of calculation for estimating individual drug clearance. AB - BACKGROUND: Carboplatin is the only cancer drug for which conventional doses are individually adjusted according to estimated clearance and target area under the curve (AUC). The aim of this prospective study was (i) to evaluate intra- and interpatient variability of ultrafilterable (UF) carboplatin AUC(0-)(infinity) and (ii) to test whether the prediction of carboplatin clearance according to the Chatelut formula established for conventional carboplatin doses was accurate for low carboplatin doses. MATERIALS AND METHODS: Thirty-one head and neck cancer patients (29 men, two women, mean age 55.9 years) received concomitant radiotherapy (Rgamma 2 Gy/day) and chemotherapy (carboplatin 50 mg/m(2)/day i.v.) for 7 weeks: Rgamma was administered 5 days/week (days 1-5) and carboplatin 2 days/week (days 1 and 4). Pharmacokinetics was performed once per week. A limited sample strategy based on Bayesian analysis was first validated and blood was subsequently taken 1 and 4 h after the end of carboplatin administration. RESULTS: A total of 143 cycles was analyzed. Ultrafilterable carboplatin AUC(0 )(infinity) ranged from 0.360 to 4.200 mg.min/ml (mean 0.830, median 0.670). As a corollary, UF carboplatin clearance ranged from 19.1 to 244.7 ml/min. Ultrafilterable carboplatin concentrations were very stable over time: AUC(0 )(infinity) variability due to treatment duration contributed to <1% of the total variance, while interpatient variability contributed to 68.6%. Accordingly, intrasubject effect was not significant (P = 0.38) whereas intersubject effect was highly significant (P <0.001). These results suggest that optimal dosage for targeting a given AUC may vary within a 13-fold range between patients. The Chatelut formula, based on creatininemia, body weight, age and sex, over estimates carboplatin clearance by 40% on average (bias 95% CI 29.6% to 51.1%). No significant relationship was observed between either bone marrow toxicity or creatinine clearance decrease and carboplatin pharmacokinetics. CONCLUSIONS: The Chatelut carboplatin clearance model established for conventional carboplatin dosages (>100 mg/m(2)) is not applicable for targeting low AUC (<1 mg x min/ml). PMID- 12649115 TI - Cancer prevalence in the UK: results from the EUROPREVAL study. AB - BACKGROUND: Cancer incidence, mortality and survival statistics for the UK are routinely available; however, data on prevalence, which is generally regarded as an important measure for health planning and resource allocation, are relatively scarce. MATERIALS AND METHODS: Eight cancer registries in the UK, covering more than half the population, provided data based on >1.5 million cases of cancer. Total prevalence was calculated using methods developed for the EUROPREVAL study, based on modelling incidence and survival trends. The prevalence of cancers of the stomach, colon, rectum, lung, breast (in females), cervix uteri, corpus uteri and prostate, melanoma of skin, Hodgkin's disease, leukaemia and all malignant neoplasms combined, was estimated for the UK for the end of 1992. RESULTS: Overall, approximately 1.5% of males and 2.5% of females in the UK population at the end of 1992 were living with a diagnosis of cancer. These proportions increased steeply with age, with approximately 7.5% (7.3% and 7.8%, in males and females, respectively) of people aged > or =65 years living with a diagnosis of cancer. Of the individual cancers, by far the highest prevalence (almost 1%) was seen for breast cancer in females; more than one in three of all living female cancer patients had been diagnosed with breast cancer. For males, around half of prevalent cases had been diagnosed >5 years previously and 30% >10 years previously; for females, these figures were both higher, at approximately 60% and 40%, respectively. CONCLUSIONS: The estimates of prevalence presented here comprise: recently diagnosed patients in need of treatment and monitoring; long term survivors, some of whom will nevertheless eventually die from the cancer, while others may be cured of the disease; and patients in the terminal phase who are dying from the cancer. Further work should attempt to identify the proportions of patients in the different phases of care in order to optimise the use of prevalence estimates in health care planning. PMID- 12649116 TI - High international prognostic score predicts a worse outcome for patients with Hodgkin's disease and HIV infection: results of a prospective study with Stanford V regimen. PMID- 12649117 TI - A case study documenting the anticancer activity of ZD1839 (Iressa) in the brain. PMID- 12649118 TI - A simple cost-effective lactate dehydrogenase level measurement can stratify patients with Ewing's tumor into low and high risk. PMID- 12649119 TI - L-Asparaginase-associated hyperlipidemia with hyperviscosity syndrome in a patient with T-cell lymphoblastic lymphoma. PMID- 12649120 TI - Sustained partial response of an intra-abdominal desmoid tumor treated with gemcitabine, 5-fluorouracil and leucovorin. PMID- 12649121 TI - Bone marrow origin of myofibroblasts in irradiation pulmonary fibrosis. AB - There is a rapid onset of organizing alveolitis/fibrosis at 120-140 d after whole lung irradiation of C57BL/6J mice. To test the hypothesis that circulating cells of bone marrow origin contribute to irradiation fibrosis, irradiated chimeric green fluorescent protein (GFP)+ C57BL/6J mice were followed for GFP+ cells in areas of lung fibrosis. In a second experimental model, C57BL/6J female mice received 20 Gy total lung irradiation, and after 60 or 80 d were intravenously injected with cells from a clonal GFP+ male bone marrow stromal cell line or male GFP+ whole bone marrow, respectively. The mice were then followed for the development of pulmonary fibrosis, and the contribution of Y-probe-positive, GFP+ cells to fibrotic areas was quantitated. Bromodeoxyuridine labeling of developing fibrotic areas showed that the cell division occurred predominantly in GFP+, Y probe-positive, and vimentin-positive cells. Immunohistochemistry demonstrated that these cells were macrophages and fibroblasts, not endothelial cells. Mice that received manganese superoxide dismutase-plasmid/liposome intratracheal injection 24 h before total lung irradiation demonstrated a decrease in GFP+ fibroblastic cells in the lung. Thus, pulmonary irradiation fibrosis contains proliferating cells of bone marrow origin, and gene therapy prevention of this condition acts in part by decreasing the migration and proliferation of marrow origin cells. PMID- 12649122 TI - In vitro surfactant protein B deficiency inhibits lamellar body formation. AB - Surfactant protein (SP) B is essential for normal pulmonary surfactant activity and lamellar body genesis in type 2 cells. However, the role of SP-B in lamellar body genesis is poorly understood. We developed an adenovirus vector expressing antisense SP-B as an alternative in vitro model of SP-B deficiency to begin to explore the role of SP-B in lamellar body genesis. RT-PCR analysis revealed that antisense SP-B expression interfered with translation of endogenous SP-B mRNA. Antisense SP-B expression resulted in reliable in vitro reproduction of many features of SP-B deficiency, including absent mature SP-B and decreased lamellar bodies and SP-C. Light and electron microscopy demonstrated significant reductions in lamellar body number. Western blotting revealed a significant reduction in mature 8-kD SP-B protein and decreased mature SP-C. Our data indicate that antisense SP-B can be effectively used to replicate the SP-B deficient type 2 cell phenotype in vitro, and provides an attractive alternative to transgenic models for the further study of the role of SP-B in lamellar body genesis. PMID- 12649123 TI - Air pollution particles produce airway wall remodeling in rat tracheal explants. AB - There is evidence that chronic exposure to high levels of ambient particulate pollutants (PM) is associated with chronic airflow obstruction, but how this occurs is not known. We exposed rat tracheal explants to Ottawa urban air particles (ECH93) or diesel exhaust particles. After 7 d in air organ culture, both types of PM increased explant procollagen and transforming growth factor (TGF)-beta 1 gene expression, and markedly increased tissue hydroxyproline. For both types of particle, nuclear factor-kappa B inhibitor SN50 completely blocked increased gene expression. With EHC93, procollagen expression was inhibited by the oxidant scavenger, tetramethylthiourea, and by the iron chelator, deferoxamine, but TGF-beta1 expression was not inhibited by deferoxamine. Inhibitors of extracellular signal regulated kinase and p38 kinase did not affect EHC93-induced gene expression. With diesel exhaust particles, tetramethylthiourea and deferoxamine had no effect, but extracellular signal regulated kinase and p38 inhibitors completely blocked effects on procollagen and TGF-beta 1. Fetuin, an inhibitor of TGF-beta receptor binding, prevented increases in procollagen gene expression. We conclude that two common types of PM can directly induce expression of genes involved in fibrogenesis and actual airway wall fibrosis through nuclear factor-kappa B- and TGF-beta-mediated mechanisms. PM-induced airway wall remodeling may play an important role in producing airflow obstruction in individuals living in high PM regions. PMID- 12649124 TI - Effect of SCH55700, a humanized anti-human interleukin-5 antibody, in severe persistent asthma: a pilot study. AB - Antagonizing the effect of interleukin (IL)-5 is a potential new treatment strategy in allergic disorders. We evaluated the safety, biological activity, and pharmacokinetics of SCH55700, a humanized anti-human IL-5 antibody, in subjects with severe persistent asthma treated with oral or high doses of inhaled steroids. In a double-blind, randomized, multicenter trial, a rising single dose of SCH55700 (0.03 mg/kg [n = 2], 0.1 mg/kg [n = 4], 0.3 mg/kg [n = 6], or 1.0 mg/kg [n = 12]) or placebo (n = 8) was administered intravenously. SCH55700 dose dependently reduced circulating eosinophil counts. At a dose of 1.0 mg/kg, the decrease remained significant up to Day 30 [(0.07 +/- 0.01) x 10(9)/L versus (0.23 +/- 0.04) x 10(9)/L at baseline] (mean +/- SEM) (p = 0.05). After administration of SCH55700 at 0.3 and 1.0 mg/kg, a trend toward improvement in baseline FEV1 was observed, which reached significance 24 hours after the 0.3 mg/kg dose (p = 0.019 versus placebo). No significant changes occurred in other clinical indices of disease activity. Adverse events were not different between active treatment and placebo. We conclude that SCH55700 is a biologically active anti-human IL-5 antibody that can be safely used in severe steroid-treated asthma. Its therapeutic potential needs to be addressed in specifically designed efficacy trials. PMID- 12649125 TI - Treatment of acute respiratory distress syndrome with recombinant surfactant protein C surfactant. AB - We performed a phase I/II trial in North America of a recombinant surfactant protein C-based surfactant (Venticute) as treatment for the acute respiratory distress syndrome. Patients were prospectively randomized to receive either standard therapy or standard therapy plus one of two doses of exogenous surfactant given four times over 24 hours. Surfactant administration was well tolerated. No significant treatment benefit was associated with surfactant treatment. Bronchoalveolar lavage of treated patients at 48 hours reflected the presence of exogenous surfactant components, did not show evidence of improved surface tension lowering function, and had interleukin-6 concentrations that were significantly lower than control group values, consistent with an antiinflammatory treatment effect. The presence of exogenous surfactant was not detected in lavage fluid obtained at 120 hours. Future studies might rationally employ larger surfactant doses and a more prolonged dosing schedule. PMID- 12649126 TI - Airflow obstruction after myeloablative allogeneic hematopoietic stem cell transplantation. AB - Despite advances in the management of myeloablative allogeneic hematopoietic stem cell transplants, airflow obstruction (AFO) remains a significant complication. We conducted a 12-year study to examine the recent epidemiology of AFO and its associated mortality. Using the rate of percent predicted FEV1 decline after transplant, we defined AFO as a more than 5% per year decline in percent predicted FEV1 with the lowest post-transplant FEV1/FVC ratio less than 0.8. New obstruction was more frequent than previous estimates (26% overall, 32% among patients with chronic graft-versus-host disease [GVHD]) and was significantly associated with older age at transplant, lower pretransplant FEV1/FVC ratio, history of both acute and chronic GVHD, and respiratory viral infection within the first 100 days after transplant. AFO was associated with significant attributable mortality rates of 9% at 3 years, 12% at 5 years, and 18% at 10 years after transplant, which were much higher for the subpopulation of patients with chronic GVHD (22% at 3 years, 27% at 5 years, and 40% at 10 years). These results suggest that the incidence of AFO may have been underestimated previously, and its presence significantly increases the mortality of long-term survivors of myeloablative allogeneic hematopoietic stem cell transplant patients. PMID- 12649128 TI - ATP stimulates the release of prostacyclin from perfused veins isolated from the hamster hindlimb. AB - ATP-stimulated prostacyclin release from veins was investigated using epigastric veins isolated from hamsters. Veins were perfused with MOPS-buffered physiological salt solution (PSS). ATP was administered into the perfusate, and the bath solution (MOPS-PSS) was collected and assayed for the presence of the stable prostacyclin metabolite 6-keto-PGF1alpha. ATP (100 microM) resulted in reproducible increases in bath concentration from 73 +/- 22 to 279 +/- 50 pg/ml (P < 0.05, n = 5). This response was abolished by indomethacin (10 microM, P < 0.05). To ascertain whether the endothelium was the source of prostacyclin, endothelium was disrupted using air (n = 10) or deoxycholic acid (n = 6). Perfusion with air significantly reduced (P < 0.05) but did not completely abolish ATP-stimulated release of prostacyclin, while deoxycholic acid totally abolished the response (P < 0.05). The nonselective P2 receptor antagonist reactive blue 2 (100 microM) attenuated ATP-mediated release of prostacyclin but did not significantly alter ACh-stimulated release of prostacyclin. The nonselective adenosine receptor antagonist xanthine amine congener (1 microM) had no effect on ATP-stimulated release, and adenosine did not stimulate the release of prostacyclin. These results show that increases in intraluminal concentration of ATP stimulate abluminal release of prostacyclin from the venous endothelium. This effect is mediated by P2 receptors while adenosine and its receptors are not involved in this response. PMID- 12649127 TI - Food-entrained circadian rhythms are sustained in arrhythmic Clk/Clk mutant mice. AB - Daily scheduled feeding is a potent time cue that elicits anticipatory activity in rodents. This food-anticipatory activity (FAA) is controlled by a food entrainable oscillator (FEO) that is distinct from light-entrained oscillators of the suprachiasmatic nucleus (SCN). Circadian rhythms within the SCN depend on transcription-translation feedback loops in which CLOCK protein is a key positive regulator. The Clock gene is expressed in rhythmic tissues throughout the brain and periphery, implicating its widespread involvement in the functioning of circadian oscillators. To examine whether CLOCK protein is also necessary for the FEO, the effect of daily food restriction was studied in homozygous Clock mutant (Clk/Clk) mice. The results show that Clk/Clk mutant mice exhibit FAA, even when their circadian wheel-running behavior is arrhythmic. As in wild-type controls, FAA in Clk/Clk mutants persists after temporal feeding cues are removed for several cycles, indicating that the FEO is a circadian timer. This is the first demonstration that the Clock gene is not necessary for the expression of a circadian, food-entrained behavior and suggests that the FEO is mediated by a molecular mechanism distinct from that of the SCN. PMID- 12649129 TI - Prognosis of inv(16)/t(16;16) acute myeloid leukemia (AML): a survey of 110 cases from the French AML Intergroup. AB - Acute myeloid leukemias (AMLs) carrying inv(16)/t(16;16) chromosomal abnormalities are associated with a good prognosis. However, studies of this AML subtype have been hampered by the few number of patients reported, frequently collectively considered with those with AML carrying the t(8;21) translocation. We performed a retrospective study in 110 patients with inv(16)/t(16;16) AML (median age, 34 years) prospectively enrolled in 6 trials conducted in France between 1987 and 1998, with the aim to investigate prognostic factors for complete remission (CR) achievement and outcome of CR patients in this AML subtype. CR rate was 93%. Bad-prognosis factors for CR achievement were higher white blood cell count (WBC) and lower platelet count (optimal cutpoints at 120 and 30 x 109/L, respectively). At 3 years, estimated overall survival, disease free survival (DFS), and cumulative incidence of relapse were 58%, 48%, and 42%, respectively. In multivariate analysis, (1) advanced age (optimal cutpoint, 35 years) was the only factor for shorter DFS and (2) advanced age and low platelet count were the 2 factors for shorter survival of CR patients. Outcome of CR patients (1) was not influenced by WBC and cytogenetic findings and (2) was similar among patients allocated to receive allogeneic transplantation, high dose, or intermediate-dose cytarabine. Interestingly, advanced age was associated with a trend for more frequent additional chromosome abnormalities and predictive of higher cumulative incidence of relapse rather than death in first CR. These results markedly contrast with those reported in patients with t(8;21) AML in whom WBC, and not age, was the main high-risk factor for relapse, DFS, and survival. PMID- 12649130 TI - NMDA receptor-mediated regulation of human megakaryocytopoiesis. AB - Identification of the regulatory inputs that direct megakaryocytopoiesis and platelet production is essential for the development of novel therapeutic strategies for the treatment of thrombosis and related hematologic disorders. We have previously shown that primary human megakaryocytes express the N-methyl-d aspartate acid (NMDA) receptor 1 (NR1) subunit of NMDA-type glutamate receptors, which appear to be pharmacologically similar to those identified at neuronal synapses, responsible for mediating excitatory neurotransmission in the central nervous system. However, the functional role of NMDA receptor signaling in megakaryocytopoiesis remains unclear. Here we provide evidence that demonstrates the fundamental importance of this signaling pathway during human megakaryocyte maturation in vitro. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of RNA extracted from CD34+-derived megakaryocytes identified expression of NR2A and NR2D receptor subunits in these cells, as well as the NMDA receptor accessory proteins, Yotiao and postsynaptic density protein 95 (PSD-95). In functional studies, addition of a selective NMDA receptor antagonist, MK-801 inhibited proplatelet formation, without affecting proliferation or apoptosis. Exposure of CD34+ cells to MK-801 cultured for 14 days in the presence of thrombopoietin induced a decrease in expression of the megakaryocyte cell surface markers CD61, CD41a, and CD42a compared with controls. At an ultrastructural level, MK-801-treated cells lacked alpha-granules, demarcated membranes, and multilobed nuclei, which were prominent in untreated mature megakaryocyte controls. Using immunohistochemistry on sections of whole tibiae from c-Mpl knockout mice we demonstrated that megakaryocytic NMDA receptor expression was maintained following c-Mpl ablation. These data support a fundamental role for glutamate signaling in megakaryocytopoiesis and platelet production, which is likely to be independent of thrombopoietin-mediated effects. PMID- 12649131 TI - Mutations in exon 2 of GATA1 are early events in megakaryocytic malignancies associated with trisomy 21. AB - Patients with Down syndrome (DS) frequently develop 2 kinds of clonal megakaryocytosis: a common, congenital, spontaneously resolving, transient myeloproliferative disorder (TMD) and, less commonly, childhood acute megakaryoblastic leukemia (AMKL). Recently, acquired mutations in exon 2 of GATA1, an X-linked gene encoding a transcription factor that promotes megakaryocytic differentiation, were described in 6 DS patients with AMKL. The mutations prevent the synthesis of the full-length GATA1, but allow the synthesis of a shorter GATA1 protein (GATA1s) that lacks the transactivation domain. To test whether mutated GATA1 is involved in the initiation of clonal megakaryoblastic proliferation or in the progression to AMKL, we screened 35 DS patients with either AMKL or TMD and 7 non-DS children with AMKL for mutations in exon 2 of GATA1. Mutations were identified in 16 of 18 DS patients with AMKL, in 16 of 17 DS patients with TMD, and in 2 identical twins with AMKL and acquired trisomy 21. Analysis revealed various types of mutations in GATA1, including deletion/insertions, splice mutations, and nonsense and missense point mutations, all of which prevent the generation of full-length GATA1, but preserve the translation of GATA1s. We also show that the likely mechanism of generation of GATA1 isoforms is alternative splicing of exon 2 rather than, or in addition to, alternative translation initiation, as was proposed before. These findings suggest that acquired intrauterine inactivating mutations in GATA1 and generation of GATA1s cooperate frequently with trisomy 21 in initiating megakaryoblastic proliferation, but are insufficient for progression to AMKL. PMID- 12649132 TI - Effective therapy for a murine model of adult T-cell leukemia with the humanized anti-CD2 monoclonal antibody, MEDI-507. AB - Adult T-cell leukemia (ATL) develops in a small proportion of individuals infected with the retrovirus human T-cell leukemia virus (HTLV-1). We evaluated the efficacy of MEDI-507 (a humanized monoclonal antibody directed against CD2) alone and in combination with humanized anti-Tac (HAT) directed toward CD25, the interleukin-2 receptor alpha (IL-2Ralpha) using a human adult T-cell leukemia xenograft model. Weekly treatments (4) with HAT significantly prolonged the survival of the ATL-bearing mice when compared with phosphate-buffered saline (PBS)-treated controls (P <.0001). Mice treated with MEDI-507 (100 microg/wk for 4 weeks) survived longer than those treated with HAT (P <.0025). Furthermore, prolonged treatment (6 months) of ATL with MEDI-507 significantly improved the outcome when compared with a short course (4 weeks) of therapy (P <.0036). Such treatment with weekly MEDI-507 for 6 months led to a prolonged survival of the ATL-bearing mice that was comparable with the survival observed in the control group of mice that did not receive a tumor or therapeutic agent. We also found that the expression of Fcgamma receptors (FcRgamma) on polymorphonuclear leukocytes and monocytes was required for MEDI-507-mediated tumor killing in vivo. Thus, the tumor-killing mechanism with MEDI-507 in vivo required the expression of the receptor FcRgammaIII on polymorphonuclear leukocytes and monocytes, suggesting that it is mediated by a form of antibody-dependent cellular cytotoxicity. These results demonstrate that MEDI-507 has therapeutic efficacy on ATL in vivo and provides support for a clinical trial involving this monoclonal antibody in the treatment of patients with CD2-expressing leukemias and lymphomas. PMID- 12649133 TI - Graft-versus-host disease following allogeneic transplantation from HLA-identical sibling with antithymocyte globulin-based reduced-intensity preparative regimen. AB - Reduced-intensity conditioning (RIC) regimens are increasingly used for allogeneic stem cell transplantation (allo-SCT). RIC has been shown to allow engraftment with minimal early transplantation-related mortality (TRM). However, in the context of RIC, predictive factors for acute and chronic graft-versus-host disease (aGVHD and cGVHD, respectively) and their effect on outcome remain unknown. In this report, we analyzed the outcome of 101 high-risk patients (70 hematologic and 31 nonhematologic malignancies) who received an HLA-identical sibling allo-SCT after RIC, including fludarabine, busulfan, and antithymocyte globulin (ATG). The cumulative incidence of grade II-IV aGVHD was 36% (95% confidence interval [CI], 27%-45%), whereas the cumulative incidence of cGVHD at 2 years was 43% (95% CI, 33%-53%). In multivariate analysis, the incidence of aGVHD was significantly associated with the ATG dose infused during conditioning (P =.0005), whereas peripheral blood as stem cell source was the only predictive factor for the development of cGVHD (P =.0007). The 1-year cumulative incidences of disease progression or relapse in patients with (n = 69) and without (n = 31) GVHD (whatever its form or grade) were 30% (95% CI, 19%-41%) and 55% (95% CI, 37% 72%), respectively (P =.02), suggesting that a potent graft-versus-tumor (GVT) effect can be achieved in high-risk patients following RIC. Moreover, the GVT effect was closely associated with GVHD without an increased risk of TRM (cumulative incidence of TRM, 18% [95% CI, 10%-25%]). Collectively, these results provide a framework for the refinement of RIC approaches designed to enhance the GVT effect with an acceptable risk of GVHD. PMID- 12649134 TI - Melphalan-mobilized blood stem cell components contain minimal clonotypic myeloma cell contamination. AB - Optimal methods of stem cell mobilization in multiple myeloma are undefined, and contaminating clonotypic cells could contribute to disease recurrence. A phase 2 trial of intravenous melphalan (60 mg/m2) and granulocyte colony-stimulating factor (G-CSF) (10 microg/kg/d) for mobilization was performed. To enhance reliability, contamination was assessed with 2 sensitive methods, immunoglobulin light and heavy chain variable region patient-specific limiting-dilution polymerase chain reaction (PCR). We evaluated 29 stem cell components (SCCs) from 15 patients; for 9 SCCs, only VL PCR was used because of light chain disease or technical problems with VH primers. For 20 SCCs, VL and VH PCR results were highly correlated (r2 = 0.93, P <.01), with 35% (7 of 20) having identical estimates. VH PCR gave significantly higher estimates for 8-and VL PCR for 5 SCCs, supporting the utility of using 2 methods. Estimated clonotypic contamination per SCC was 0.0009% (range, 0%-0.1%) or 0.5 x 10(4) clonotypic cells per kilogram (range, 0-41.2 x 10(4)/kg), and contamination correlated with CD34+ cells collected (r2 = 0.42, P <.01). Melphalan-mobilized SCCs contain minimal clonotypic contamination. PMID- 12649135 TI - ATP gradients inhibit the migratory capacity of specific human dendritic cell types: implications for P2Y11 receptor signaling. AB - Dendritic cells (DCs) are specialized antigen-presenting cells residing in tissues, from which they take up antigen. Activated DCs migrate through chemokine gradients from sites of inflammation to lymph nodes to stimulate T cells. At sites of inflammation, nucleotides, such as adenosine triphosphate (ATP), are released by activated or dying cells and can function as signaling molecules through P2 receptors (P2Rs). We investigated P2R expression in different DC populations and the effect of nucleotides on chemokine-directed migration. Exposure of monocyte-derived DCs (MoDCs) and CD1a+ dermal DCs to gradients of ATP inhibited their migratory capacity in a dose-dependent manner. Studies using P2R agonists and antagonists implicated signaling through the P2Y11R. On maturation, MoDCs down-regulated P2Y11R expression and were less sensitive to ATP-mediated inhibition of migration. In contrast, ATP did not inhibit the migration of CD1c+ peripheral blood (PB) DCs or interleukin-3 receptor-positive (IL-3R+) plasmacytoid DCs. Although all 4 DC populations expressed mRNA for P2Y11R, calcium-flux studies showed that blood DC types were unresponsive to P2Y11R agonists. In conclusion, DCs use distinct subtypes of P2R. The formation of ATP gradients at sites of inflammation may transiently inhibit the migration of local DCs, thus prolonging the time of antigen encounter. P2R inhibition may represent a new strategy to improve the migration of antigen-loaded DCs from the vaccination site to lymph nodes. PMID- 12649137 TI - Depsipeptide (FR901228) induces histone acetylation and inhibition of histone deacetylase in chronic lymphocytic leukemia cells concurrent with activation of caspase 8-mediated apoptosis and down-regulation of c-FLIP protein. AB - Depsipeptide is in clinical trials for chronic lymphocytic leukemia (CLL) on the basis of earlier observations demonstrating selective in vitro activity in CLL. We sought to determine the relationship of histone H3 and H4 acetylation, inhibition of histone deacetylase, and apoptosis observed in CLL cells to justify a pharmacodynamic end point in these clinical trials. We demonstrate that in vitro depsipeptide induces histone H3 and H4 acetylation and histone deacetylase enzyme inhibition at concentrations corresponding to the LC50 (concentration producing 50% cell death) for cultured CLL cells (0.038 microM depsipeptide). The changes in histone acetylation are lysine specific, involving H4 K5, H4 K12, and H3 K9, and to a lesser extent H4 K8, but not H4 K16 or H3 K14. Depsipeptide induced apoptosis is caspase dependent, selectively involving the tumor necrosis factor (TNF) receptor (extrinsic pathway) initiating caspase 8 and effector caspase 3. Activation of caspase 8 was accompanied by the down-regulation of cellular FLICE-inhibitory protein (c-FLIP, I-FLICE) without evidence of Fas (CD95) up-regulation. Changes in other apoptotic proteins, including Bcl-2, Bax, Mcl-1, and X-linked inhibitor of apoptosis (XIAP), were not observed. Our results demonstrate a relationship between target enzyme inhibition of histone deacetylase, histone H3 and H4 acetylation, and apoptosis involving the TNF receptor pathway of apoptosis that is not used by other therapeutic agents in CLL. These data suggest use of histone H3 and H4 acetylation, inhibition of histone deacetylase, and down-regulation of FLIP as pharmacodynamic end points for further evaluation of this drug in patients. PMID- 12649136 TI - Transgenic delivery of VEGF to mouse skin leads to an inflammatory condition resembling human psoriasis. AB - Gene therapy approaches involving vascular endothelial growth factor (VEGF) to promote therapeutic angiogenesis are under consideration for conditions ranging from ischemic heart disease to nonhealing skin ulcers. Here we make the surprising observation that the transgenic delivery of VEGF to the skin results in a profound inflammatory skin condition with many of the cellular and molecular features of psoriasis, including the characteristic vascular changes, epidermal alterations, and inflammatory infiltrates. Even longstanding psoriatic disease remains dependent on the transgenic VEGF in this model because it can be effectively reversed by the addition of VEGF Trap, a potent VEGF antagonist. Previous attempts to faithfully replicate the psoriatic phenotype through the transgenic delivery of epidermal keratinocyte growth factors or inflammatory mediators generated phenotypes with only partial resemblance to human psoriasis, leaving unanswered questions about the etiology of this disease. The ability of transgenic VEGF to induce a psoriasiform phenotype suggests a new etiology and treatment approach for this disease and further substantiates emerging concerns about possible proinflammatory adverse effects that might be associated with therapeutic attempts to deliver VEGF. PMID- 12649138 TI - Defect in neutrophil killing and increased susceptibility to infection with nonpathogenic gram-positive bacteria in peptidoglycan recognition protein-S (PGRP S)-deficient mice. AB - Insect peptidoglycan recognition protein-S (PGRP-S), a member of a family of innate immunity pattern recognition molecules conserved from insects to mammals, recognizes bacterial cell wall peptidoglycan and activates 2 antimicrobial defense systems, prophenoloxidase cascade and antimicrobial peptides through Toll receptor. We show that mouse PGRP-S is present in neutrophil tertiary granules and that PGRP-S-deficient (PGRP-S-/-) mice have increased susceptibility to intraperitoneal infection with gram-positive bacteria of low pathogenicity but not with more pathogenic gram-positive or gram-negative bacteria. PGRP-S-/- mice have normal inflammatory responses and production of tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6). Neutrophils from PGRP-S-/- mice have normal phagocytic uptake of bacteria but are defective in intracellular killing and digestion of relatively nonpathogenic gram-positive bacteria. Therefore, mammalian PGRP-S functions in intracellular killing of bacteria. Thus, only bacterial recognition by PGRP-S, but not its effector function, is conserved from insects to mammals. PMID- 12649139 TI - Platelet-collagen interaction: is GPVI the central receptor? AB - At sites of vascular injury, platelets come into contact with subendothelial collagen, which triggers their activation and the formation of a hemostatic plug. Besides glycoprotein Ib (GPIb) and alphaIIbbeta3 integrin, which indirectly interact with collagen via von Willebrand factor (VWF), several collagen receptors have been identified on platelets, most notably alpha2beta1 integrin and the immunoglobulin (Ig) superfamily member GPVI. Within the last few years, major advances have been made in understanding platelet-collagen interactions including the molecular cloning of GPVI, the generation of mouse strains lacking individual collagen receptors, and the development of collagen receptor-specific antibodies and synthetic peptides. It is now recognized that platelet adhesion to collagen requires prior activation of integrins through "inside-out" signals generated by GPVI and reinforced by released second-wave mediators adenosine diphosphate (ADP) and thromboxane A2. These developments have led to revision of the original "2-site, 2-step" model, which now places GPVI in a central position in the complex processes of platelet tethering, activation, adhesion, aggregation, degranulation, and procoagulant activity on collagen. This review discusses these recent developments and proposes possible mechanisms for how GPVI acts in concert with other receptors and signaling pathways to initiate hemostasis and arterial thrombosis. PMID- 12649140 TI - Dual effects of macrophage inflammatory protein-1alpha on osteolysis and tumor burden in the murine 5TGM1 model of myeloma bone disease. AB - Recent data have implicated macrophage inflammatory protein-1alpha (MIP-1alpha) in multiple myeloma (MM)-associated osteolysis. However, it is unclear whether the chemokine's effects are direct, to enhance osteolysis, or indirect and mediated through a reduction in tumor burden, or both. It is also unclear whether MIP-1alpha requires other factors such as receptor activator of nuclear factor kappaB ligand (RANKL) for its effects on bone. In murine 5TGM1 (Radl) myeloma bearing mice, administration of neutralizing anti-MIP-1alpha antibodies reduced tumor load assessed by monoclonal paraprotein titers, prevented splenomegaly, limited development of osteolytic lesions, and concomitantly reduced tumor growth in bone. To determine the effects of MIP-1alpha on bone in vivo, Chinese hamster ovary (CHO) cells secreting human MIP-1alpha (CHO/MIP-1alpha) were inoculated into athymic mice. Mice bearing intramuscular CHO/MIP-1alpha tumors developed lytic lesions at distant skeletal sites, which occurred earlier and were larger than those in mice with CHO/empty vector (EV) tumors. When experimental metastases were induced via intracardiac inoculation, mice bearing CHO/MIP-1alpha tumors developed hypercalcemia and significantly more osteolytic lesions than mice bearing CHO/EV tumors, with intramedullary CHO/MIP-1alpha tumors associated with significantly more tartrate-resistant acid phosphatase-positive (TRAP+) osteoclasts. Injection of recombinant MIP-1alpha over calvariae of normal mice evoked a striking increase in osteoclast formation, an effect dependent on RANK/RANKL signaling because MIP-1alpha had no effect in RANK-/- mice. Together, these results establish that MIP-1alpha is sufficient to induce MM-like destructive lesions in bone in vivo. Because, in the 5TGM1 model, blockade of osteoclastic resorption in other situations does not decrease tumor burden, we conclude that MIP-1alpha exerts a dual effect in myeloma, on osteoclasts, and tumor cells. PMID- 12649141 TI - PECAM-1 functions as a specific and potent inhibitor of mitochondrial-dependent apoptosis. AB - Programmed cell death, or apoptosis, is a tightly regulated, naturally occurring process by which damaged or unwanted cells are removed. Dysregulated apoptosis has been implicated in a variety of pathophysiological conditions, including degenerative diseases, tissue remodeling, and tumorigenesis. The decision to live or die results from integration of numerous environmental signals transmitted by specific classes of cell surface receptors that bind hormones, growth factors, or components of the extracellular matrix. Here we show that platelet endothelial cell adhesion molecule-1 (PECAM-1), a homophilic-binding member of the immunoreceptor tyrosine-based inhibitory motif (ITIM) family of inhibitory receptors, functions prominently to inhibit apoptosis in naturally occurring vascular cells subjected to apoptotic stimuli. Murine endothelial cells and human T lymphocytes lacking PECAM-1 were found to be far more sensitive than their PECAM-1-expressing counterparts to multiple death signals that stimulate Bax, a multidomain, proapoptotic member of the Bcl-2 family that plays a central role in mitochondrial dysfunction-dependent apoptosis. In addition, PECAM-1 markedly suppressed Bax overexpression-induced cytochrome c release, caspase activation, and nuclear fragmentation. Amino acid substitutions within PECAM-1's extracellular homophilic binding domain, or within its cytoplasmic ITIM, completely abolished PECAM-1-mediated cytoprotection. Taken together, these data implicate PECAM-1 as a novel and potent suppressor of Bax-mediated apoptosis and suggest that members of the immunoglobulin gene (Ig) superfamily, like cell surface integrins, may also transmit survival signals into blood and vascular cells. PMID- 12649142 TI - IVIg-mediated amelioration of murine ITP via FcgammaRIIB is independent of SHIP1, SHP-1, and Btk activity. AB - It has been established that amelioration of murine immune thrombocytopenia purpura (ITP) by IVIg is dependent on the inhibitory receptor FcgammaRIIB. Co cross-linking of the FcgammaRIIB with the B-cell receptor complex or with FcepsilonRI in mast cells results in cell inhibition, which is mediated by recruitment of the inositol phosphatase SHIP1 to the cytoplasmic tail of the FcgammaR. The FcgammaRIIB can also associate with protein tyrosine phosphatase SHP-1 as a potential secondary target of the receptor. Alternatively, homoaggregation of FcgammaRIIB can induce a proapoptotic state in B cells that is dependent on the presence of Bruton tyrosine kinase (Btk), a kinase also expressed in monocytes. We sought to determine if these signaling pathways may direct IVIg-mediated FcgammaRIIB-dependent regulation of in vivo monocyte function in a murine model of ITP in which IVIg functions in an FcgammaRIIB dependent manner. We demonstrate that mice deficient in SHIP1, SHP-1, and Btk respond to the ameliorating effects of IVIg with the same kinetics as control mice. We conclude that IVIgmediated inhibitory pathways operating via monocyte FcgammaRIIB may involve a transmembrane signaling pathway different from that of B cells. PMID- 12649143 TI - Immunogenicity and protective efficacy of orally administered recombinant Lactococcus lactis expressing surface-bound HIV Env. AB - This study investigates whether genetically modified orally administered Lactococcus lactis (L lactis) could be used as an HIV vaccine. L lactis is immunogenic and extremely safe when delivered orally. We created a recombinant L lactis vector expressing the envelope protein of HIV on its cell surface. Oral immunization with this vector induced high levels of HIV-specific serum IgG and fecal IgA antibodies. Cell-mediated immune responses also were generated in both the regional lymph nodes and the spleen. Dendritic cells are readily infected by L lactis and appear to play a potential role in mediating the development of these immune responses. The protective efficacy of this vaccine strategy was demonstrated by challenging mice intraperitoneally with an HIV Env-expressing vaccinia virus. Their viral loads were 350-fold lower than those of control mice. These findings support the further development of L lactis-based HIV vaccines. PMID- 12649144 TI - Founder von Willebrand factor haplotype associated with type 1 von Willebrand disease. AB - To date, no dominant mutation has been identified in a significant proportion of patients with type 1 von Willebrand disease (VWD). In this study, we examined 70 families as part of the Canadian Type 1 VWD Study. The entire VWF gene was sequenced for 1 index case, revealing 2 sequence variations: intron 30 (5312 19A>C) and exon 28 at Tyr1584Cys (4751A>G). The Tyr1584Cys variation was identified in 14.3% (10 of 70) of the families and was in phase with the 5312 19A>C variation in 7 (10.0%) families. Both variants were observed in 2 of 10 UK families with type 1 VWD, but neither variant was found in 200 and 100 healthy, unrelated persons, respectively. Mean von Willebrand factor antigen (VWF:Ag), VWF ristocetin cofactor (VWF:RCo), and factor VIII coagulant activity (FVIII:C) for the index cases in these families are 0.4 U/mL, 0.36 U/mL, and 0.54 U/mL, respectively, and VWF multimer patterns show no qualitative abnormalities. Aberrant VWF splicing was not observed in these patients, and both alleles of the VWF gene are expressed as RNA. Molecular dynamic simulation was performed on a homology model of the VWF-A2 domain containing the Tyr1584Cys mutation. This showed that no significant structural changes occur as a result of the substitution but that a new solvent-exposed reactive thiol group is apparent. Expression studies revealed that the Tyr1584Cys mutation results in increased intracellular retention of the VWF protein. We demonstrate that all the families with the Tyr1584Cys mutation share a common, evolved VWF haplotype, suggesting that this mutation is ancient. This is the first report of a mutation that segregates in a significant proportion of patients with type 1 VWD. PMID- 12649145 TI - Comparative response of plasma VWF in dogs to up-regulation of VWF mRNA by interleukin-11 versus Weibel-Palade body release by desmopressin (DDAVP). AB - Recombinant human interleukin-11 (rhIL-11), a glycoprotein 130 (gp130)-signaling cytokine approved for treatment of thrombocytopenia, also raises von Willebrand factor (VWF) and factor VIII (FVIII) by an unknown mechanism. Desmopressin (1 deamino-8-d-arginine vasopressin [DDAVP]) releases stored VWF and FVIII and is used for treatment of VWF and FVIII deficiencies. To compare the effect of these 2 agents, heterozygous von Willebrand disease (VWD) and normal dogs were treated with either rhIL-11 (50 microg/kg/d subcutaneously x 7 days) or DDAVP (5 microg/kg/d intravenously x 7 days). The rhIL-11 produced a gradual and sustained elevation of VWF and FVIII levels in both heterozygous VWD and normal dogs while DDAVP produced a rapid and unsustained increase. Importantly, rhIL-11 treatment produced a 2.5- to 11-fold increase in VWF mRNA in normal canine heart, aorta, and spleen but not in homozygous VWD dogs, thus identifying a mechanism for elevation of plasma VWF in vivo. Moreover, dogs pretreated with rhIL-11 retain a DDAVP-releasable pool of VWF and FVIII, suggesting that rhIL-11 does not significantly alter trafficking of these proteins to or from storage pools. The half-life of infused VWF is unchanged by rhIL-11 in homozygous VWD dogs. These results show that rhIL-11 and DDAVP raise plasma VWF by different mechanisms. Treatment with rhIL-11 with or without DDAVP may provide an alternative to plasma derived products for some VWD and hemophilia A patients if it is shown safe in clinical trials. PMID- 12649146 TI - Human T lymphocytes transduced by lentiviral vectors in the absence of TCR activation maintain an intact immune competence. AB - Gene transfer into T lymphocytes is currently being tested for the treatment of lymphohematologic disorders. We previously showed that suicide gene transfer into donor lymphocytes infused to treat leukemic relapse after allogeneic hematopoietic stem cell transplantation allowed control of graft-versus-host disease. However, the T-cell receptor (TCR) activation and sustained proliferation required for retroviral vector transduction may impair the half life and immune competence of transduced cells and reduce graft-versus-leukemia activity. Thus, we tested lentiviral vectors (LVs) and stimulation with cytokines involved in antigen-independent T-cell homeostasis, such as interleukin 7 (IL-7), IL-2, and IL-15. Late-generation LVs transduced efficiently nonproliferating T cells that had progressed from G0 to the G1 phase of the cell cycle on cytokine treatment. Importantly, IL-2 and IL-7, but not IL-15, stimulation preserved physiologic CD4/CD8 and naive-memory ratios in transduced cells with only minor induction of some activation markers. Functional analysis of immune response to cytomegalovirus (CMV) showed that, although CMV-specific T cells were preserved by all conditions of transduction, proliferation and specific killing of autologous cells presenting a CMV epitope were higher for IL-2 and IL-7 than for IL-15. Thus, LV transduction of IL-2 or IL-7 prestimulated cells overcomes the limitations of retroviral vectors and may significantly improve the efficacy of T cell-based gene therapy. PMID- 12649147 TI - Tolerance induction to a mammalian blood group-like carbohydrate antigen by syngeneic lymphocytes expressing the antigen, II: tolerance induction on memory B cells. AB - Induction of immune tolerance on memory B cells specific to transplantation carbohydrate antigens was studied in the experimental animal model of alpha1,3galactosyltransferase knockout (KO) mice, which lack the alpha-gal epitope (Galalpha1-3Galbeta1-4GlcNAc-R) and can produce the anti-Gal antibody against it. Memory anti-Gal B cells were generated by immunization of KO mice with pig kidney membranes (ie, xenogeneic cell membranes expressing an abundance of alpha-gal epitopes). Lymphocytes including memory anti-Gal B cells were administered into lethally irradiated KO mice, together with syngeneic wild-type (WT) lymphocytes expressing alpha-gal epitopes. Memory anti-Gal B cells were completely tolerized after being in vivo for 14 days with WT lymphocytes. This was indicated by the lack of anti-Gal immunoglobulin G (IgG) response following immunization with pig kidney membranes vs the extensive anti-Gal response in mice that did not receive WT lymphocytes. Tolerance induction was prevented if T cells were activated by alloantigens. This tolerance was highly specific to anti-Gal B cells and did not affect memory B cells with closely related specificity, such as B cells with anti-blood group A specificity. Tolerance induction on anti-Gal B cells was found to be time dependent and required more than 10 days of in vivo exposure of these B cells to WT lymphocytes. These observations suggest a novel method for induction of tolerance to transplantation carbohydrate antigens in humans, by in vitro transduction of autologous blood lymphocytes with an adenovirus containing the corresponding glycosyltransferase gene and administration of the transduced cells into the circulation after removal of natural antibodies to the antigen. PMID- 12649148 TI - Accelerated senescence of human erythrocytes cultured with Plasmodium falciparum. AB - Red blood cells infected withPlasmodium falciparum(IRBCs) undergo changes primarily in their membrane composition that contribute to malaria pathogenesis. However, all manifestations (eg, anemia) cannot be accounted for by IRBCs alone. Uninfected erythrocytes (URBCs) may play a role, but they have been under researched. We wanted to document changes in the erythrocyte membrane that could contribute to URBC reduced life span and malaria-associated anemia. Human erythrocytes were cultured withP falciparumand washed at the trophozoite stage. IRBCs and URBCs were separated on Percoll density gradient, thus obtaining erythrocyte fractions of different densities/ages. IRBC- and URBC-purified membranes were analyzed and compared with control normal erythrocytes (NRBCs) of the same age, from the same donor, kept in the same conditions.P falciparumaccelerated aging of both IRBCs and URBCs, causing a significant shift in the cell population toward the denser (old) fraction. Protein, phospholipid, and cholesterol content were reduced in IRBCs and young URBCs. Young and medium uninfected fractions had higher levels of lipid peroxidation and phospholipid saturation (because of the loss of polyunsaturated fatty acids, PUFAs) and lower phosphatidylserine. In IRBCs, thiobarbituric reactive substances (TBARSs) were higher, and PUFAs and phosphatidylserine lower than in NRBCs and URBCs. In comparison, trophozoite membranes had lower phospholipid (particularly sphingomyelin and phosphatidylserine) and cholesterol content and a higher degree of saturation. Parasite-induced peroxidative damage might account for these modifications. In summary, we demonstrated that membrane damage leading to accelerated senescence of both infected and uninfected erythrocytes will likely contribute to malaria anemia. PMID- 12649149 TI - Selective blockade of CD28 and not CTLA-4 with a single-chain Fv-alpha1 antitrypsin fusion antibody. AB - B7-1 and B7-2 are costimulatory molecules expressed on antigen-presenting cells. The CD28/B7 costimulation pathway is critical for T-cell activation, proliferation, and Th polarization. Blocking both cytotoxic T-lymphocyte associated antigen 4 (CTLA-4) and CD28 interactions with a CTLA-4/Ig fusion protein inhibits various immune-mediated processes in vivo, such as allograft rejection and autoimmunity. However, selective blockade of CD28 may represent a better strategy for immunosuppression than B7 blockade, because CTLA-4/B7 interactions have been shown to participate in the extinction of the T-cell receptor-mediated activation signal and to be required for the induction of immunologic tolerance. In addition, selective CD28 inhibition specifically decreases the activation of alloreactive and autoreactive T cells, but not the activation of T cells stimulated by exogenous antigens presented in the context of self major histocompatibility complex (MHC) molecules. CD28 blockade cannot be obtained with anti-CD28 dimeric antibodies, which cluster their target and promote T-cell costimulation, whereas monovalent Fab fragments can block CD28 and reduce alloreactivity. In this study, we report the construction of a monovalent single-chain Fv antibody fragment from a high-affinity antihuman CD28 antibody (CD28.3) that blocked adhesion of T cells to cells expressing the CD28 receptor CD80. Genetic fusion with the long-lived serum protein alpha1-antitrypsin led to an extended half-life without altering its binding characteristics. The anti-CD28 fusion molecule showed biologic activity as an immuno-suppressant by inhibiting T cell activation and proliferation in a mixed lymphocyte reaction. PMID- 12649150 TI - Myeloid and monocytic dyspoiesis as determined by flow cytometric scoring in myelodysplastic syndrome correlates with the IPSS and with outcome after hematopoietic stem cell transplantation. AB - Marrow cells of myeloid lineage from 115 patients with myelodysplastic syndrome (MDS) were characterized by multidimensional flow cytometry and compared with findings in 104 patients with various disorders and 25 healthy donors. Based on phenotypic and scatter characteristics, a flow cytometric scoring system (FCSS) was developed that allowed for a simple numerical display of results. The flow cytometric scores were categorized as normal/mild (0-1), moderate (2-3), or severe (> or = 4). Most flow cytometric abnormalities were significantly (P <.05) more frequent in patients with MDS than in the control cohort. Flow cytometric scores in MDS patients were then retrospectively compared with marrow blast counts assessed by morphology, cytogenetics, hematologic parameters, and International Prognostic Scoring System (IPSS) risk categorization. The flow cytometric scores correlated inversely with leukocyte and absolute neutrophil counts (P <.01) and correlated directly with IPSS scores (P <.01) and with IPSS cytogenetic risk categories (P <.01). In 111 MDS patients who underwent allogeneic hematopoietic stem cell transplantation, flow scores correlated with posttransplantation outcome. The probabilities of posttransplantation relapse were 3%, 15%, and 33% for patients with mild, moderate, and severe FCSS scores, respectively (P <.01), and overall survival was 74%, 40%, and 36%, respectively, for the 3 groups (P <.01). In multivariate analyses, there was a significant contribution of the flow score independent of the IPSS in predicting survival and relapse (P <.01, P =.02, and P =.03, respectively). These data suggest that FCSS is useful in assessing marrows for diagnosis of MDS and in determining the prognostic outcome in patients with this disorder. PMID- 12649151 TI - Asp1424Asn MYH9 mutation results in an unstable protein responsible for the phenotypes in May-Hegglin anomaly/Fechtner syndrome. AB - May-Hegglin anomaly (MHA), Fechtner syndrome (FTNS), Sebastian syndrome (SBS), and Epstein syndrome (EPS) are a group of rare, autosomal dominant disorders characterized by thrombocytopenia, giant platelets, and Dohle-like inclusion bodies, together with variable manifestations of Alport-like symptoms that include high-tone sensorineural deafness, cataracts, and nephritis. These disorders result from mutations in the MYH9 gene, which encodes for the nonmuscle myosin heavy chain A protein (also known as NMMHC-A). To date 20 different mutations have been characterized for this gene, but no clear phenotype-genotype correlation has been established, and very little is known regarding the molecular pathogenesis of this group of diseases. Here, we describe 2 new families with MHA/FTNS phenotypes that have been characterized in terms of their mutations, protein localization in megakaryocytes, protein expression, and mRNA stability. Our findings suggest that, at least for the Asp1424Asn mutation in the MYH9 gene, the phenotypes result from a highly unstable protein. No abnormalities in protein localization or mRNA stability were observed. We hypothesize that haploinsufficiency of the MYH9 results in a failure to properly reorganize the cytoskeleton in megakaryocytes as required for efficient platelet production. PMID- 12649152 TI - Very late relapse in diffuse large B-cell lymphoma represents clonally related disease and is marked by germinal center cell features. AB - Patients with diffuse large B-cell lymphoma (DLBCL) rarely show relapse after 4 years of complete remission (CR). In this study, we addressed the following questions: (1) Does late-relapsing DLBCL represent clonally related disease or a second malignancy; and (2) is there a characteristic biologic background? In 10 of 13 DLBCL patients with relapse after 4 to 17 years, a clonal relationship was established based on identical IgH-sequences and/or identical bcl2-IgH translocation. Most (77%) showed features of germinal center (GC) cells, as defined by expression of CD10, bcl-2, and bcl-6 protein and ongoing immunoglobulin heavy chain variable region (VH) hypermutation. A GC phenotype was seen in 8 (20%) of 38 control patients matched for age, stage, and (extra)nodal localization with relapse within 2.5 years (P =.005). In conclusion, we have found evidence that late-relapsing DLBCL represents truly clonally related disease episodes in most cases and that this clinical behavior may be related to the biologic features of GC cells. PMID- 12649153 TI - Lack of the CD8+ cell anti-HIV factor in CD8+ cell granules. AB - In HIV infection, CD8+ cells show cytotoxic and noncytotoxic anti-HIV activity. The latter function is mediated, at least in part, by a secreted antiviral protein, the CD8+ cell antiviral factor (CAF). Because antiviral effector molecules, such as perforin and granzymes, reside in the exocytic granules of CD8+ T cells, we examined the possibility that granules contain CAF-like activity. CD8+ cells from HIV-infected individuals showing strong CAF-mediated antiviral activity were induced to release their granule constituents into culture media. Within 1 hour of stimulation, high levels of granzyme B (a primary granule constituent) were found in the culture fluids of previously activated CD8+ cells. The same culture fluids contained no or very low amounts of CAF activity, as measured with HIV-infected CD4+ cells. Maximal levels of CAF activity were not observed until 5 or 7 days after stimulation, consistent with typical CAF production kinetics. In addition, extracts of granules purified from antiviral CD8+ cells did not show any CAF activity, whereas the cytoplasmic fraction of these cells showed substantial levels of antiviral activity. These findings suggest that CAF does not reside at appreciable levels in the exocytic granules of antiviral CD8+ T cells. PMID- 12649154 TI - Multiparity induces priming to male-specific minor histocompatibility antigen, HY, in mice and humans. AB - One of the factors that increases the risk of graft-versus-host disease following allogeneic stem cell transplantation is the use of multiparous females as donors. Since minor histocompatibility (H) antigens are the main targets of graft-versus host and graft-versus-leukemia responses, we tested the hypothesis that multiparity could prime minor H antigen-specific T cells. We examined the peripheral lymphoid populations of multiparous mice and humans for evidence of priming of CD8+ T-cytotoxic lymphocytes against peptide epitopes of the male specific minor H antigen, HY. In contrast to naive females, multiparous females have measurable levels of circulating HY-specific tetramer-positive T lymphocytes, which can be readily expanded in vitro. These findings have implications for the in vitro generation of T-cell clones as reagents for immunotherapy for tumors following stem cell transplantation. PMID- 12649155 TI - CD8alpha+ mouse spleen dendritic cells do not originate from the CD8alpha- dendritic cell subset. AB - Although previous studies had indicated that the CD8alpha- and CD8alpha+ subtypes of murine dendritic cells (DCs) differ in immediate origin, a recent study found that intravenous transfer of CD8alpha- DCs led to CD8alpha+ DCs in the spleen several days later, suggesting a direct precursor-product relationship. We have repeated these experiments with a balance sheet approach. We find that though a few CD8alpha+ DCs can be generated in such experiments, this is a rare event and could be the result of a contaminant precursor. Most of the immediate precursors of CD8alpha+ DCs are cells that lack the phenotype of a recognizable DC. CD8alpha DCs and CD8alpha+ DCs are not precursor-product related, though these sublineages may be connected further upstream. PMID- 12649156 TI - Proangiogenic properties of human myeloma cells: production of angiopoietin-1 and its potential relationship to myeloma-induced angiogenesis. AB - Patients with multiple myeloma (MM) have increased bone marrow (BM) angiogenesis; however, the proangiogenic properties of myeloma cells and the mechanisms of MM induced angiogenesis are not completely clarified. The angiopoietin system has been identified as critical in the regulation of vessel formation. In this study we have demonstrated that myeloma cells express several proangiogenic factors, and, in particular, we found that angiopoietin-1 (Ang-1), but not its antagonist Ang-2, was expressed by several human myeloma cell lines (HMCLs) at the mRNA and the protein levels. In a transwell coculture system, we observed that myeloma cells up-regulated the Ang-1 receptor Tie2 in human BM endothelial cells. Moreover, in an experimental model of angiogenesis, the conditioned medium of HMCLs significantly stimulated vessel formation compared with control or vascular endothelial growth factor (VEGF) treatment. The presence of anti-Tie2 blocking antibody completely blunted the proangiogenic effect of XG-6. Finally, our in vitro results were supported by the in vivo finding of Ang-1, but not Ang-2, mRNA and protein expression in purified MM cells obtained from approximately 47% of patients and by high BM angiogenesis in patients with MM positive for Ang-1, suggesting that the angiopoietin system could be involved, at least in part, in MM-induced angiogenesis. PMID- 12649157 TI - Functional phenotype of phosphoinositide 3-kinase p85alpha-null platelets characterized by an impaired response to GP VI stimulation. AB - Phosphoinositide 3-kinases (PI3Ks), a family of lipid kinases comprising 3 classes with multiple isoforms, have been shown to participate in different phases of platelet signaling. To investigate the roles that enzymes play in platelet function in vivo and determine which isoforms are important for particular signaling events, we analyzed platelet function of gene knockout mice deficient in the p85alpha regulatory subunit of heterodimeric class IA PI3K. The kinase activity of p85alpha-/- platelets was only 5% of the activity of platelets from wild-type littermates. Platelet aggregation induced by adenosine diphosphate (ADP), thrombin, U46619, phorbol 12-myristate 13-acetate (PMA), or botrocetin was not defective in p85alpha-/- mice, compared with wild-type animals. In contrast, aggregation induced by collagen and collagen-related peptide (CRP) was partially but readily impaired in p85alpha-/- mice. Both P-selectin expression and fibrinogen binding in response to CRP were also decreased to a similar extent in p85alpha-/- platelets. Platelets from p85alpha-/- mice appeared to spread poorly over a CRP-coated surface with intact filopodial protrusions. Significant attenuation of CRP-induced tyrosine phosphorylation in known PI3K effectors such as Btk, Tec, PKB/Akt, and phospholipase Cgamma2 were observed with p85alpha-/- platelets, whereas no alteration was noted in upstream molecules of Syk, LAT, and SLP-76. Considered as a whole, these results provide the first genetic evidence that PI3K p85alpha plays a significant role in platelet function, almost exclusively in the glycoprotein (GP) VI/Fc receptor gamma chain complex-mediated signaling pathway. PMID- 12649158 TI - Genetically targeted radiotherapy for multiple myeloma. AB - Multiple myeloma is a disseminated neoplasm of terminally differentiated plasma cells that is incurable with currently available therapies. Although the disease is radiosensitive, external beam radiation leads to significant toxicity due to sensitive end-organ damage. Thus, genetic approaches for therapy are required. We hypothesized that the incorporation of immunoglobulin promoter and enhancer elements in a self-inactivating (SIN) lentiviral vector should lead to specific and high-level transgene expression in myeloma cells. A SIN lentivector with enhanced green fluorescent protein (EGFP) expression under the control of a minimal immunoglobulin promoter as well as the Kappa light chain intronic and 3' enhancers transduced myeloma cell lines with high efficiency (30%-90%). EGFP was expressed at a high level in myeloma cells but silent in all nonmyeloma cell lines tested compared with the cytomegalovirus (CMV) promoter/enhancer. Transduction of myeloma cells with the targeted vector coding for the human sodiumiodide symporter (hNIS) led to hNIS expression by these cells allowing them to concentrate radioiodine up to 18-fold compared with controls. Tumor xenografts in severe combined immunodeficiency mice expressing hNIS could be imaged using iodine-123 (123I) and shown to retain iodide for up to 48 hours. These tumor xenografts were completely eradicated by a single dose of the therapeutic isotope iodine-131 (131I) without evidence of recurrence up to 5 months after therapy. We conclude that lentivectors can be transcriptionally targeted for myeloma cells and the use of hNIS as a therapeutic gene for myeloma in combination with 131I needs further exploration. PMID- 12649159 TI - Impact of CD34+ cell dose on the outcome of patients undergoing reduced-intensity conditioning allogeneic peripheral blood stem cell transplantation. AB - We analyzed the impact of CD34+ cell dose on the outcome of 86 patients undergoing reduced-intensity conditioning (RIC) allogeneic peripheral blood stem cell transplantation. The RIC was based on fludarabine 150 mg/m2 and melphalan 140 mg/m2 or busulphan 10 mg/kg. A median of 5.68 x 106 CD34+ cells/kg and 2.86 x 108 CD3+ cells/kg were infused. All patients receiving more than percentile 75 (p75) of CD34+ cells reached complete chimerism in T lymphocytes by days 21 to 28, compared with 44% among those receiving p75 or fewer cells (P =.046). Overall, 30.3% patients developed grade 2 to 4 acute graft-versus-host disease (aGVHD). Among 83 evaluable patients, 55.8% developed chronic GVHD (cGVHD). The dose of CD34+ cells infused did influence the development of cGVHD, with a cumulative incidence of extensive cGVHD of 74% vs 47% (P =.02) among patients receiving more than p75 CD34+ cells vs those receiving p75 or fewer. Projected overall survival (OS) and event-free survival (EFS) at 43 months were 60% and 46%, respectively. Neither the dose of CD34+ cells nor the dose of CD3+ cells infused significantly influenced OS and EFS, although among patients categorized as high-risk, 36% of those receiving p75 or fewer CD34+ cells relapsed or progressed, compared with only 9% among those receiving more than p75 CD34+ cells (P =.07). Among patients receiving p75 or fewer CD34+ cells, 36% of high-risk patients relapsed, compared with 10% of low- and intermediate-risk patients (P =.004), while relapse rates were not significantly different between both subgroups when we infused more than p75 CD34+ cells, thus indicating that infusing high doses of CD34+ cells ameliorates the negative effect of advanced disease status at transplantation. cGVHD was associated with better EFS (63% vs 16% at 43 months for patients with and without cGVHD; P <.0001) and better OS (78% vs 28% for patients with and without cGHVD; P <.001). The number of CD34+ cells infused should be tailored to prevent extensive cGVHD among patients categorized as low-risk, while high-risk patients, in whom the graft-versus leukemia effect may determine disease outcome, should receive high doses of CD34+ cells. PMID- 12649160 TI - Fanconi anemia protein complex: mapping protein interactions in the yeast 2- and 3-hybrid systems. AB - Fanconi anemia (FA) is an autosomal recessive syndrome characterized by progressive bone marrow failure and cancer predisposition. Eight FA complementation groups have been identified. The FANCA, FANCC, FANCE, FANCF, and FANCG proteins form a nuclear complex required for the monoubiquination of the FANCD2 protein. To investigate the architecture of the FA protein complex, the yeast 2-hybrid system was used to map contact points of the FANCA/FANCG, FANCC/FANCE, and FANCF/FANCG interactions. FANCG was shown to interact with both the amino-terminus of FANCA and the carboxyl-terminal region of FANCF. A FANCG mutant truncated at the carboxyl-terminus retained the ability to interact with FANCA. The interaction between FANCG and FANCF was ablated by a Leu71Pro mutant of FANCG. A central region of FANCE was sufficient for FANCC binding. A Leu554Pro mutant of FANCC failed to interact with FANCE. To further examine complex assembly, the yeast 3-hybrid system was used to investigate the ability of FANCG to act as a molecular bridge in mediating interaction between other FA proteins. FANCG was able to mediate interaction between FANCA and FANCF, as well as between monomers of FANCA. Direct interaction between FANCE and FANCD2 was also demonstrated in the yeast 2-hybrid system. This interaction involving an amino terminal region of FANCD2 may provide a link between the FA protein complex and its downstream targets. PMID- 12649161 TI - The heme-heme oxygenase system: a molecular switch in wound healing. AB - When cells are injured they release their contents, resulting in a local accumulation of free heme proteins and heme. Here, we investigated the involvement of heme and its degrading enzyme heme oxygenase (HO) in the inflammatory process during wound healing. We observed that heme directly accumulates at the edges of the wound after inflicting a wound in the palate of Wistar rats. This coincided with an increased adhesion molecule expression and the recruitment of leukocytes. To prove that heme is responsible for the recruitment of leukocytes, heme was administered intradermally 24 hours prior to injury. A clear heme-induced influx of both macrophages and granulocytes was observed. When examining the HO isoforms, HO-1 and HO-2, we found that HO-2 was present in the entire submucosa. Surprisingly, we observed also that HO-1 is significantly expressed in the epithelium of both the mucosa and the skin of animals without wounds. On inflammation, HO-1 expression increased, particularly in infiltrating cells during the resolution phase of inflammation. Interestingly, we observed that heme-induced influx of leukocytes was highly elevated after pharmacologic inhibition of HO activity. These observations suggest that the heme HO system is closely involved in the control of wound healing. Our results demonstrate that the local release of heme may be a physiologic trigger to start inflammatory processes, whereas HO-1 antagonizes inflammation by attenuating adhesive interactions and cellular infiltration. Moreover, the basal level of HO expression in the skin may serve as a first protective environment against acute oxidative and inflammatory insults. PMID- 12649162 TI - Red cell adenylate kinase deficiency: molecular study of 3 new mutations (118G>A, 190G>A, and GAC deletion) associated with hereditary nonspherocytic hemolytic anemia. AB - We report here 2 patients with chronic nonspherocytic hemolytic anemia (CNSHA) and severe red blood cell (RBC) adenylate kinase (AK) deficiency. One of these patients, a boy of Spanish origin, exhibited a neonatal icterus and splenomegaly and required blood transfusions until the age of 2 years. The other patient was a white, American infant born to parents who were first cousins; he also presented with neonatal icterus and anemia. In neither case was psychomotor impairment observed. The first patient was found to be a compound heterozygote for 2 different missense mutations, 118G>A(Gly40Arg) and 190G>A(Gly64Arg) (cDNA sequence first described by Matsuura et al, 1989). The second patient was homozygous for an in-frame deletion (GAC) from nucleotide (nt) 498 to 500 or nt 501 to 503 of the cDNA sequence, predicting deletion of either aspartic acid (Asp) 140 or 141. The crystal structure of porcine cytosolic AK was used as a molecular model to investigate how these mutations may affect enzyme structure and function. PMID- 12649163 TI - SU5416, a small molecule tyrosine kinase receptor inhibitor, has biologic activity in patients with refractory acute myeloid leukemia or myelodysplastic syndromes. AB - Increased bone marrow angiogenesis and vascular endothelial growth factor (VEGF) levels are adverse prognostic features in patients with acute myeloid leukemia (AML) or myelodysplastic syndromes (MDSs). VEGF is a soluble circulating angiogenic molecule that stimulates signaling via receptor tyrosine kinases (RTKs), including VEGF receptor 2 (VEGFR-2). AML blasts may express VEGFR-2, c kit, and FLT3. SU5416 is a small molecule RTK inhibitor (RTKI) of VEGFR-2, c-kit, and both wild-type and mutant FLT3. A multicenter phase 2 study of SU5416 was conducted in patients with refractory AML or MDS. For a median of 9 weeks (range, 1-55 weeks), 55 patients (33 AML: 10 [30%] primary refractory, 23 [70%] relapsed; 22 MDS: 15 [68%] relapsed) received 145 mg/m2 SU5416 twice weekly intravenously. Grade 3 or 4 drug-related toxicities included headaches (14%), infusion-related reactions (11%), dyspnea (14%), fatigue (7%), thrombotic episodes (7%), bone pain (5%), and gastrointestinal disturbance (4%). There were 11 patients (20%) who did not complete 4 weeks of therapy (10 progressive disease, 1 adverse event); 3 patients (5%) who achieved partial responses; and 1 (2%) who achieved hematologic improvement. Single agent SU5416 had biologic and modest clinical activity in refractory AML/MDS. Overall median survival was 12 weeks in AML patients (range, 4-41 weeks) and not reached in MDS patients. Most observed toxicities were attributable to drug formulation (polyoxyl 35 castor oil or hyperosmolarity of the SU5416 preparation). Studies of other RTKI and/or other antiangiogenic approaches, with correlative studies to examine biologic effects, may be warranted in patients with AML/MDS. PMID- 12649164 TI - Sorting of soluble TNF-receptor for granule storage in hematopoietic cells as a principle for targeting of selected proteins to inflamed sites. AB - Hematopoietic cells have secretory lysosomes that degranulate at the inflammatory site upon stimulation. We asked whether one could target exogenous proteins with a therapeutic potential to secretory lysosomes in hematopoietic cells. For this purpose, we expressed a soluble tumor necrosis factor (TNF) receptor form (sTNFR1) in hematopoietic cell lines. In order to accomplish targeting to secretory lysosomes, both endoplasmic reticulum (ER) retention and constitutive secretion have to be prevented. ER export was facilitated by addition of a transmembrane (tm) sequence, and constitutive secretion was overcome by incorporating a cytosolic sorting signal (Y) from CD63. This signal directed the resulting sTNFR1-tm-Y to secretory lysosomes. Confirmation of these results was provided by biosynthetic radiolabeling, subcellular fractionation, immunofluorescence microscopy, and immunoelectron microscopy. The tm-Y fragment was cleaved by proteolysis, resulting in generation of the membrane-free sTNFR1 in secretory lysosomes. Our results suggest a potential for using the storage organelles of hematopoietic cells as vehicles for targeting sites of inflammation with therapeutically active agents. PMID- 12649165 TI - A 6-month versus a 12-month surveillance for hepatocellular carcinoma in 559 hemophiliacs infected with the hepatitis C virus. AB - Hepatocellular carcinoma (HCC) is an increasingly frequent cause of mortality in hemophiliacs with chronic viral hepatitis. Early diagnosis of the tumor at an initial stage is known to improve the outcome of HCC treatment. Because all HCC cases detected in a previous study based upon annual ultrasound (US) surveillance of hemophiliacs with elevated alanine aminotransferase levels were multinodular, this study was designed to evaluate if a more intense surveillance with US and alphafetoprotein (AFP) serum levels of all the patients infected with the hepatitis C virus (HCV) improved the identification of single nodule tumors. A multicenter cohort of 559 HCV-infected hemophiliacs was divided into 2 arms, one followed up at 6-month intervals and one at 12-month intervals depending on the choice and available facilities of each treatment center. During a 6-year surveillance period, HCC was diagnosed in 5 (2.4%) of 210 patients in the 6-month group and in 3 (0.9%) of 349 patients in the 12-month group. The overall incidence rate of HCC was 239 per 100 000 per year (397 per 100 000 per year in the 6-month group and 143 per 100 000 per year in the 12-month group; differences not statistically significant). By multivariate analysis, HCC risk was increased 12.9-fold with alcohol intake more than 80 g/d and 15.2-fold with AFP levels higher than 11 ng/mL. Liver-related death occurred in 8 cases (1.4%), including 3 with HCC. Still alive and tumor free after 24 to 34 months from diagnosis are 3 patients with multinodular tumors treated with repeat chemoembolization followed by orthotopic liver transplantation. In conclusion, 6-month surveillance with US did not increase the chances of detection of single nodule tumors, but it is reasonable to assume that successful treatment of multinodular tumors based upon debulking with chemoembolization and liver transplantation was facilitated by this approach. PMID- 12649166 TI - Therapeutic effect of idiotype-specific CD4+ T cells against B-cell lymphoma in the absence of anti-idiotypic antibodies. AB - Immunoglobulin (Ig) variable (V) region idiotypes (Id's) are highly tumor specific antigens produced by B-lymphoma cells and are promising targets for immunotherapy. Id vaccination has proven effective in experimental mouse models and may possibly prevent recurrence of B lymphomas in humans. It has previously been shown that anti-Id antibodies protect against B-cell lymphoma in the absence of T cells. We here demonstrate in a T-cell-receptor transgenic mouse model that the contrary is also true: Id-specific CD4+ T cells can protect against Id+ B lymphoma cells in the absence of B cells, antibodies, and CD8+ T cells. Moreover, Id-specific CD4+ T cells have a curative potential since they could be transferred as late as 17 days after subcutaneous tumor cell injection of severe combined immunodeficiency (SCID) mice and still abrogate tumor development in about 50% of mice. Such mice undergo an acute inflammatory swelling with infiltration of neutrophils at the site of tumor injection, which subsides over weeks, with some mice cured and delayed emergence of lymphomas in other mice. Adoptively transferred CD4+ T cells accumulated in the tumor and were activated (CD69+). In vitro experiments demonstrated that memory, but not naive, Id specific CD4+ T cells kill Id+ B-lymphoma cells. The results show that Id specific CD4+ T cells, in the absence of antibodies home to subcutaneous Id+ B lymphoma, become activated, induce inflammation, and prevent tumor development. PMID- 12649167 TI - Phospholipid scramblase 3 is the mitochondrial target of protein kinase C delta induced apoptosis. AB - Protein kinase C (PKC) delta translocates to mitochondria during apo-ptosis,but its mitochondrial target remains unclear. We found that PKC-delta physically interacted with and phosphorylated phospholipid scramblase 3 (PLS3) after UV irradiation. PLS3 is a high-affinity substrate for PKC-delta in vitro with the K(m) at 10.5 nM. Cells expressing wild-type PLS3 became apoptotic on phorbol ester stimulation, whereas the control cells did not. Expression of a mitochondrial-targeted PKC-delta enhanced apoptosis more prominently in HeLa-PLS3 cells than control HeLa cells and HeLa cells expressing an inactive PLS3 mutant. These results indicate that PLS3 is a downstream effector of PKC-delta in the mitochondria. PMID- 12649168 TI - Death receptor 4 and bladder cancer risk. AB - Tumor necrosis factor-related apoptosis-inducing ligand stimulates the extrinsic apoptoticpathway by binding to death receptors 4 (DR4) and 5 (DR5). In DR4 exon 4, a C-->G polymorphism at amino acid 626 located immediately 3' to one of the main receptor ligand interface regions, results in a threonine-->arginine change. We found that the DR4 exon 4 G/G genotype was associated with an overall decreased risk of bladder cancer in Caucasians [odds ratio (OR) = 0.58; 95% confidence interval (CI), 0.38-0.88]. This protective effect was more apparent in younger individuals (OR = 0.42; 95% CI, 0.20-0.87) than in older individuals (OR = 0.60; 95% CI, 0.35-1.02) and in women (OR = 0.45; 95% CI, 0.20-0.99) than in men (OR = 0.60; 95% CI, 0.36-0.99). Moreover, the protective effect was greater for light smokers (OR = 0.19; 95% CI, 0.06-0.59) compared with heavy smokers (OR = 0.83; 95% CI, 0.41-1.69). These data provide the first large-scale molecular epidemiological evidence that the DR4 polymorphism is associated with environmental exposure and bladder cancer risk, possibly through modulating the capacity of the receptor ligand complex to engage the apoptotic pathway. PMID- 12649169 TI - Optical bioluminescence and positron emission tomography imaging of a novel fusion reporter gene in tumor xenografts of living mice. AB - Noninvasive imaging of reporter gene expression using various imaging modalitiesis playing an increasingly important role in defining molecular events in the field of cancer biology, cell biology, and gene therapy. In this study, a novel reporter vector was constructed encoding a fusion protein comprised of a mutant herpes simplex virus type 1 thymidine kinase (HSV1-sr39tk) (tk), a positron emission tomography (PET) reporter gene, and renilla luciferase (rl), a bioluminescence optical reporter gene joined by a 20 amino acid long spacer sequence. We validated the activity of the two enzymes encoded by the fusion protein (tk(20)rl) in cell culture. Then, tumors stably expressing the tk(20)rl fusion gene were imaged both by microPET and optically using a cooled charge coupled device camera in xenograft-bearing living mice. Using a single fusion reporter (PET/optical) gene should accelerate the validation of reporter gene approaches developed in cell culture for translation into preclinical and clinical models. PMID- 12649171 TI - Distinct amplification of an untranslated regulatory sequence in the egfr gene contributes to early steps in breast cancer development. AB - Overexpression of the epidermal growth factor receptor (egfr) gene is a common feature in breast cancer. We demonstrated recently that the expression of EGFR in breast cancer strongly correlates with the length of a CA simple sequence repeat within the first 2000 bases in intron 1 of the egfr gene [CA simple sequence repeat (CA-SSR) I; H. Buerger et al., Cancer Res., 60: 854-857, 2000]. Using a standardized semiautomated method of microsatellite analysis for loss of heterozygosity detection, we identified an allelic imbalance (AI) at the egfr locus in 55 of 163 primary breast cancer cases. Fine mapping of the chromosomal region at 7p12-15 around the egfr gene using 10 CA-SSR markers showed that mutations of egfr in breast cancer are frequently restricted to the first intron of egfr. Thereby, the simple sequence repeat CA-SSR I in intron 1 was affected in 84% of the patients with AI. Reverse transcription-PCR analysis of 23 breast cancer tissues with AI excluded the presence of in-frame deletions between exon 2 and exon 7. For additional characterization of the underlying phenomenon leading to the detection of an AI in microsatellite analysis, a quantitative 5'-nuclease assay for the first CA-SSR I in intron 1 was established. In breast cancer cases with AI the presence of amplifications of this sequence was shown. Kaplan-Meier analysis revealed a statistically significant worse prognosis for patients with AI in the cancer tissue at the egfr locus compared with patients without AI. Interestingly, 75% of the patients bearing AI of CA-SSR I in the tumor also showed AI at normal, nontumorous breast tissue. Our data strongly support the assumption that distinct amplifications in intronic sequences of the egfr gene, which enhance the basic transcription activity of the gene, represent one of the first steps in breast carcinogenesis. Furthermore, they point to the presence of prognosis-associated markers for breast cancer already in morphological normal breast tissue. PMID- 12649170 TI - Vascular endothelial growth factor C expression and lymph node metastasis are regulated by the type I insulin-like growth factor receptor. AB - Vascular endothelial growth factor (VEGF)-C is a lymphangiogenic factor implicated in lymphatic metastasis. In this study, we investigated the role of the type I insulin-like growth factor receptor (IGF-IR) in the regulation of VEGF C expression. We used Lewis lung carcinoma subline M-27 cells transfected with human IGF-IR cDNA. These cells, but not the wild-type cells, expressed VEGF-C mRNA, produced a M(r) 58,000 VEGF-C precursor protein, and secreted a M(r) 29,000 processed form in response to IGF-I. In vivo, they acquired a lymph node metastasizing potential. VEGF-C induction was abolished in cells expressing an IGF-IR with tyrosine-phenylalanine substitutions in the kinase domain, but not in the COOH-terminal domain. The induction was phosphatidylinositol 3'-kinase dependent and, to a lesser extent, mitogen-activated protein kinase signaling dependent, as determined by the use of the respective inhibitors LY294002 (84.6% reduction) and PD98059 (38% reduction). The results identify the IGF-IR as a positive regulator of VEGF-C expression and implicate it in the control of lymphatic metastasis. PMID- 12649172 TI - Tissue microarray analysis reveals site-specific prevalence of oncogene amplifications in head and neck squamous cell carcinoma. AB - Fluorescence in situ hybridization was applied on a collection of 609 squamous cell carcinomas of the head and neck (HNSCCs),including 511 primary carcinomas of different clinical stage and anatomical localization and 98 recurrent carcinomas, second primary carcinomas, and regional metastases on a tissue microarray. The overall prevalence of amplifications of five oncogenes analyzed was 34.5% for CCND1, 12.7% for EGFR, 8.8% for MYC, 6.2% for ZNF217, and 3.6% for ERBB2. CCND1 amplifications were associated with the pharyngeal site in primary carcinomas (P < 0.001), whereas amplifications of ZNF217 were less frequent in pharyngeal carcinomas as compared with primary oral and laryngeal carcinomas (P = 0.02). The amplification pattern of these oncogenes suggests that different molecular pathways are involved in HNSCCs of different localizations. PMID- 12649173 TI - Restoration of fragile histidine triad (FHIT) expression induces apoptosis and suppresses tumorigenicity in breast cancer cell lines. AB - The fragile histidine triad (FHIT) gene at chromosome 3p14.2 is a tumor suppressor gene that is altered mainly by deletion in a large fraction of human tumors, including breast cancers. To evaluate the potential of FHIT gene therapy in this type of cancer, we have studied the biological effects of adenoviral FHIT transduction (Ad-FHIT) in breast cancer cell lines. The results showed that, after FHIT restoration in BT-549, MDA-MB-436, and HCC1806 cells, they underwent apoptosis by activation of the intrinsic pathway. In all three cell lines infected with Ad-FHIT, we have found activation of caspase-2, which is required for permeabilization of mitochondria, release of cytochrome c, and apoptosis. Furthermore, Fhit overexpression produces alteration in cell cycling properties, as well as reduction of the tumorigenic potential in nude mice. PMID- 12649174 TI - Is the p53 inactivation frequency in squamous cell carcinomas of the head and neck underestimated? Analysis of p53 exons 2-11 and human papillomavirus 16/18 E6 transcripts in 123 unselected tumor specimens. AB - Mutations and interaction with high-risk human papillomavirus (HPV) E6 oncoprotein are well-established mechanisms of p53 inactivation. In a series of 123 unselected squamous cell carcinomas of the head and neck (SCCHN), we performed sequence analysis of the entire coding region of p53 transcript and determined the presence of the E6 transcripts of HPV 16 and 18. Aberrant p53 transcripts were identified in 97 (79%) SCCHN. HPV 16 and/or 18 E6 transcripts were detected in 37 (30%) tumor specimens, including 20 (77%) of the 26 p53 wild type tumors. The likely inactivation of p53 in 117 (95%) of the 123 SCCHN suggests that this event could be obligatory in the multistep process of carcinogenesis. PMID- 12649175 TI - A Nodal- and ALK4-independent signaling pathway activated by Cripto-1 through Glypican-1 and c-Src. AB - Human Cripto-1 (CR-1) is a member of the epidermal growth factor-Cripto FRL1 Cryptic family that has been shown to function as a coreceptor with the type I Activin serine-threonine kinase receptor ALK4 for the transforming growth factor beta-related peptide Nodal. However, CR-1 can also activate the mitogen-activated protein kinase and Akt pathways independently of Nodal and ALK4 by an unknown mechanism. Here, we demonstrate that CR-1 specifically binds to Glypican-1, a membrane-associated heparan sulfate proteoglycan, and activates the tyrosine kinase c-Src, triggering the mitogen-activated protein kinase and Akt signaling pathways. Finally, an active Src kinase is necessary for CR-1 to induce in vitro transformation and migration in mouse mammary epithelial cells. PMID- 12649176 TI - Threonines 2638/2647 in DNA-PK are essential for cellular resistance to ionizing radiation. AB - DNA-dependent protein kinase (DNA-PK) is required for the repair of double stranded DNA breaks through the nonhomologous DNA end joining pathway. DNA-PK activity is required for DNA repair, but kinase activity also appears to be attenuated through an autoregulatory feedback loop. We show that autophosphorylation of DNA-PK catalytic subunit occurs in trans at least three sites NH(2) terminal to the catalytic domain and that two sites, threonine 2638 and 2647, determine DNA-PK autophosphorylation in vitro. Thr2638/2647ala substitution in DNA-PK catalytic subunit compromised cellular resistance to ionizing radiation without affecting DNA end joining, suggesting a requirement for DNA-PK inactivation for cell survival at a step after the rejoining of double stranded DNA breaks. PMID- 12649177 TI - A novel gene, MSI2, encoding a putative RNA-binding protein is recurrently rearranged at disease progression of chronic myeloid leukemia and forms a fusion gene with HOXA9 as a result of the cryptic t(7;17)(p15;q23). AB - The pathogenetic role of the P210 BCR/ABL1 fusion gene in the chronic phase of chronic myeloid leukemia (CML) has been well established.In contrast, the genetic mechanisms underlying the disease progression into the accelerated phase (AP) and the final blast crisis (BC) remain poorly understood. We have previously identified (A. Barbouti et al., Genes Chromosomes Cancer, 35: 127-137, 2002) two cryptic balanced translocations, t(7;17)(p15;q23) and t(7;17)(q32-34;q23), in CML AP/BC using multicolor fluorescence in situ hybridization. In this study, we show that a novel gene in 17q23, Musashi-2 (MSI2), encoding a putative RNA-binding protein, is rearranged in both cases and that a MSI2/HOXA9 fusion gene is formed in the case with the 7p15 breakpoint. The identified in-frame MSI2/HOXA9 fusion transcript retains both of the RNA recognition motif domains of MSI2, which is fused to the homeobox domain of HOXA9, and is likely to play an important role in the disease progression of CML. PMID- 12649179 TI - Dicoumarol: a unique microtubule stabilizing natural product that is synergistic with Taxol. AB - In studies on the antiproliferative actions of coumarin compounds, we discovered that dicoumarol (a coumarin anticoagulant; 3,3'-methylenebis[4-hydroxycoumarin]) inhibits the first cleavage of Strongylocentrotus purpuratus (sea urchin) embryos in a concentration-dependent manner with 50% inhibition occurring at a concentration of 10 microM. Because first cleavage in sea urchin embryos is highly selective for microtubule-targeted agents, we thought that the active compounds might inhibit cell division by interacting with tubulin or microtubules. We found that dicoumarol binds to bovine brain tubulin with a K(d) of 22 microM and that 0.1 microM dicoumarol strongly stabilizes the growing and shortening dynamics at the plus ends of the microtubules in vitro. Dicoumarol reduces the rate and extent of shortening, it increases the percentage of time the microtubules spend in an attenuated (paused) state, and it reduces the overall dynamicity of the microtubules. The antimitotic effects of the widely used cancer chemotherapeutic agent Taxol (paclitaxel) are also mediated by suppressing microtubule dynamics. We demonstrate that exposure to combinations of Taxol and dicoumarol results in a synergistic inhibition of cell division of sea urchin embryos. The results suggest that the antiproliferative mechanism of action of dicoumarol and possibly related pharmacophores may be mediated by tubulin binding and the stabilization of spindle microtubule dynamics. Because of its low toxicity and simple chemical structure, there is potential interest to explore combinations of antimitotic coumarins with other chemotherapeutic agents to improve efficacy and lower toxicity. PMID- 12649178 TI - Elevated levels of microtubule destabilizing factors in a Taxol resistant/dependent A549 cell line with an alpha-tubulin mutation. AB - The A549 Taxol-resistant cell lines, A549-T12 and A549-T24, were isolated in our laboratory, and are dependent on Taxol for normal growth. The microtubules in these cells displayed increased dynamicity in the absence of Taxol. In the present study, a heterozygous point mutation in Kalpha1-tubulin was discovered at alpha379 (Ser to Ser/Arg). Although Taxol binds to beta-tubulin in the microtubule, sequencing of beta-tubulin class I did not reveal any mutations. The expression of the alpha-tubulin mutation was demonstrated using high-resolution isoelectric focusing and two-dimensional gel analysis. Both the wild-type and mutant tubulin were expressed in the Taxol-resistant cell lines. The region of alpha-tubulin that encompasses alpha379 is near the COOH terminus that has been proposed as a site of interaction with microtubule-associated protein (MAP) 4 and stathmin, a tubulin-interacting protein. In the Taxol-resistant cells, the active nonphosphorylated form of stathmin was increased approximately 2-fold, whereas the inactive phosphorylated forms were barely detected. The inactive phosphorylated forms of MAP4 were increased in the A549-T12 and A549-T24 cell lines. We hypothesize that these changes in tubulin/MAPs that result in increased microtubule instability may be related to the alpha-tubulin mutation and are compensated for by the stabilizing properties of Taxol. PMID- 12649180 TI - Inhibition of proliferation and estrogen receptor signaling by peroxisome proliferator-activated receptor gamma ligands in uterine leiomyoma. AB - Peroxisome proliferator-activated receptor (PPAR) gamma is an important signaling molecule in cells of mesenchymal origin, inducing differentiation and regulating cell proliferation in several cell types such as vascular smooth muscle cells. Leiomyomas arise from smooth muscle cells of the uterine myometrium with an incidence rate as high as 70% in women of reproductive age. PPAR signaling has not been characterized in these tumors, although prostaglandins, natural PPAR ligands, are known effectors of key biological functions in the normal myometrium. Leiomyomas and tumor-derived cells isolated from a rat model for this disease were characterized by Western analysis and found to express all three PPAR isoforms, suggesting that signaling pathways mediated by these receptors were intact in this tumor type. In vitro experiments with a leiomyoma-derived cell line demonstrated that the pan-PPAR ligand cis-4,7,10,13,16,19 docosahexaenoic acid and PPARgamma-specific ligands 15-deoxy-delta(12,14) prostaglandin J(2), troglitazone, and ciglitazone inhibited 17beta-estradiol stimulated cell proliferation. This inhibitory effect was not observed with PPARalpha- or PPARbeta-specific ligands. Although both PPAR and estrogen receptor (ER) signaling pathways were intact in leiomyoma cells, in addition to growth inhibition, stimulation of PPARgamma signaling also inhibited ER-mediated gene expression. Human leiomyomas were also found to express all three PPAR isoforms, and primary cultures of these cells were sensitive to the inhibitory effects of PPARgamma ligands. These results suggest that in uterine leiomyomas PPARgamma activation is growth inhibitory and that this inhibition is mediated at least in part by negative cross-talk between ER and PPAR signaling pathways. PMID- 12649181 TI - Effect of energy restriction on cell cycle machinery in 1-methyl-1-nitrosourea induced mammary carcinomas in rats. AB - Energy restriction (ER) results in a profound inhibition of chemically induced mammary carcinogenesis. The cancer inhibitory activity of ER has been shown to be associated with lower rates of cell proliferation during both premalignant and malignant stages of this disease process. Moreover, inhibition of carcinogenesis and suppression of cell proliferation occur in animals in which plasma concentrations of insulin-like growth factor (IGF)-I are reduced, and plasma corticosterone levels are increased concomitantly. Given the role of both hormones in signal transduction pathways that can modulate cell cycle progression, albeit via different regulatory mechanisms, we report experiments conducted to determine whether hypothesized effects of changes in plasma levels of IGF-I and corticosterone on cell cycle regulation could be detected in mammary carcinomas occurring in 40% ER rats in comparison to ad libitum fed control rats or 40% ER rats that were energy repleted for 7 days (ER-REP). As determined by appropriate combinations of immunoprecipitations, Western blots, and kinase activity assays, it was found that levels of phosphorylated retinoblastoma and E2F-1 were significantly reduced by ER (approximately 40 and 75%, respectively; P < 0.01), an effect that was partially reversed by ER-REP. Reductions in cyclin dependent kinase (CDK)2 (82%) and CDK4 (77%) kinase activity in ER carcinomas were likely to account for the observed effects on retinoblastoma and E2F-1. Both Cip1/p21 and Kip1/p27 and levels of these proteins complexed with CDK2 were significantly elevated in ER carcinomas (P < 0.01), and levels of cyclin E were reduced. On the other hand, regulation of CDK4 kinase activity by ER was likely attributable to effects on cyclin D1 as well as increased binding of P16 and P19 to CDK4. The majority of changes induced by ER were reversed by ER-REP. These observations are consistent with the hypothesis that ER exerts its profound cancer inhibitory activity, in part, by multifaceted regulation of cell cycle machinery, possibly via concomitant changes in corticosterone and IGF-1 metabolism, although the role of other hormones and growth factors should not be dismissed. PMID- 12649182 TI - Cyclin E expression is a significant predictor of survival in advanced, suboptimally debulked ovarian epithelial cancers: a Gynecologic Oncology Group study. AB - Cyclin E is a key regulator of the G(1)-S transition. Abnormalities in cyclin E expression have been related to survival in a variety of cancers. This study evaluated the prognostic relevance of cyclin E in human ovarian cancer. Immunohistochemical expression of cyclin E was evaluated in 139 advanced, suboptimally debulked epithelial ovarian cancer specimens from patients treated on Gynecologic Oncology Group protocol 111. High cyclin E protein expression (> or =40% cyclin E positive tumor cells) was seen in 62 (45%) of the advanced, suboptimally debulked ovarian cancer patients. Expression of cyclin E was not associated with age, race, stage, grade, cell type, or amount of residual disease. High verses low cyclin E expression was associated with a shorter median survival (29 +/- 2 versus 35 +/- 3 months) and worse overall survival (P < 0.05). Univariate and multivariate regression analyses revealed that high relative to low cyclin E was associated with a 40-50% increase in the risk of death (hazard rate, P < or = 0.05). Fluorescence in situ hybridization was used in a subset of 20 cases to examine cyclin E gene amplification. Eight of 10 cases with high cyclin E expression exhibited amplification of the cyclin E gene, whereas only 1 of 10 cases with low expression displayed gene amplification (P < 0.006). High cyclin E expression was an independent poor prognostic factor for patients with advanced ovarian cancer, and it was associated with amplification of the cyclin E gene. PMID- 12649183 TI - Functional blocks in caspase activation pathways are common in leukemia and predict patient response to induction chemotherapy. AB - Defects in apoptosis mechanisms contribute to chemoresistance in malignancy. However, correlations of apoptosis-regulating proteins with clinical outcome in cancer patients are variable, presumably reflecting the difficulty of using static tests of gene expression in a scenario influenced by a dynamic interplay of multiple pro- and antiapoptotic molecules. Therefore, we assessed the functional integrity of apoptosis pathways in intact primary leukemia cells and correlated the functional status of these pathways with clinical outcome. Active apoptogenic proteins were introduced into primary leukemia cells by electroporation followed by measurement of active caspases by flow cytometric techniques. Cytochrome c was introduced to activate the intrinsic (mitochondrial) pathway, whereas caspase-8 was introduced to activate the extrinsic (death receptor) pathway. In a series of 24 patients with acute myeloid leukemia, 79% had a block in at least one pathway, indicating that defects in caspase activation mechanisms are common in patients with leukemia. Simultaneous blocks in both pathways correlated with chemoresistant disease (92% of patients with chemoresistant disease versus 33% of patients with chemosensitive disease; P = 0.005) and decreased overall patient survival (35% versus 89% 1-year survival; P = 0.02). Simultaneous blockage of the intrinsic and extrinsic pathways could be explained by a defect located at a point of convergence of the two pathways, probably related to overexpression of endogenous inhibitors of the effector caspases, rather than decreased levels of these proteases. This study supports the importance of apoptosis pathways in determining response to chemotherapy and suggests that functional defects in caspase activation are prognostic in patients with leukemia. PMID- 12649184 TI - A common variant of the methylenetetrahydrofolate reductase gene (1p36) is associated with an increased risk of cancer. AB - Folate metabolism is thought to play an important role in carcinogenesis through its involvement in both DNA methylation and nucleotide synthesis. A common Ala(222)/Val variant in the methylenetetrahydrofolate reductase (MTHFR) gene leads to a disturbed folate metabolism and is associated with decreased genomic DNA methylation. We previously reported that the MTHFR Val/Val genotype was associated with increased cancer mortality in men from a population-based cohort of subjects ages > or = 85 years. To further explore the deleterious effects of the MTHFR genotype, we studied the association of the genotype with cancer risk in 860 men ages 65-84 years who were followed >10 years (Zutphen Elderly Study). During follow-up, 149 new cases of cancer occurred among the 793 men without cancer at baseline. The risk of developing cancer was 1.80-fold (95% confidence interval, 1.09-3.00) higher among men with the Val/Val genotype than among men with the Ala/Ala genotype. Except for lung cancer [relative risk (RR), 1.15], the risks of common forms of cancers were significantly increased among men with the Val/Val genotype [cancer of the prostate (RR, 3.48); the colorectum (RR, 3.65); the kidney and bladder (RR, 5.48)]. The risks of cancer were particularly increased among men with a lower folate and a higher alcohol intake and men of an older age. In conclusion, our current and previous studies in two independent populations indicate that a common Ala/Val variant in the MTHFR gene is a risk factor for cancer in elderly men from the general population. The mechanism underlying this association might involve genomic instability as a result of insufficient methylation of genomic DNA. PMID- 12649185 TI - Caspase-8 and apoptosis-inducing factor mediate a cytochrome c-independent pathway of apoptosis in human colon cancer cells induced by the dietary phytochemical chlorophyllin. AB - Chlorophyllin (CHL), an antimutagenic and anticarcinogenic water-soluble derivative of chlorophyll, was recently found to be highly effective as a chemopreventive agent in a high-risk population exposed unavoidably to aflatoxin B(1) in the diet (P. A. Egner et al., Proc. Natl. Acad. Sci. USA, 98: 14601 14606, 2001). The current study examined the response of HCT116 human colon cancer cells to CHL treatment. Cells exposed to concentrations in the range 0.0625-0.5 mM CHL underwent growth arrest and apoptosis after 24 h, with the formation of a sub-G(1) peak in the attached cell population and nuclear condensation in the floating cell population. There was a concentration-dependent attenuation of mitochondrial membrane potential (deltapsi(m)) without the release of cytochrome c or activation of the caspase-9/caspase-3/poly(ADP-ribose) polymerase pathway. However, apoptosis-inducing factor was released from mitochondria into the cytosol and translocated to the nucleus, leading to concentration-dependent cleavage of nuclear lamins. The upstream mediators of this CHL-induced apoptosis pathway were identified as caspase-8/caspase-6 and truncated Bid, acting in conjunction with other proapoptotic members of the Bcl-2 family, such as Bak. These findings suggest that CHL might trigger apoptosis via interaction with putative "death receptors" in the plasma membrane of cancer cells, leading to initial cleavage of procaspase-8 and activation of subsequent downstream events, resulting in the destruction of nuclear lamins. Importantly, E cadherin and alkaline phosphatase, which are indicators of cell differentiation, were strongly induced at all concentrations of CHL. Thus, in addition to being an effective blocking agent during the initiation phase, these findings support a role for CHL as a suppressing agent and as a possible novel therapeutic strategy directed toward aberrant cell proliferation in the colon. PMID- 12649187 TI - Discovery of JSI-124 (cucurbitacin I), a selective Janus kinase/signal transducer and activator of transcription 3 signaling pathway inhibitor with potent antitumor activity against human and murine cancer cells in mice. AB - Constitutively activated, tyrosine-phosphorylated signal transducer and activator of transcription (STAT) 3 plays a pivotal role in human tumor malignancy. To discover disrupters of aberrant STAT3 signaling pathways as novel anticancer drugs, we developed a phosphotyrosine STAT3 cytoblot. Using this high throughput 96-well plate assay, we identified JSI-124 (cucurbitacin I) from the National Cancer Institute Diversity Set. JSI-124 suppressed the levels of phosphotyrosine STAT3 in v-Src-transformed NIH 3T3 cells and human cancer cells potently (IC(50) value of 500 nM in the human lung adenocarcinoma A549) and rapidly (complete inhibition within 1-2 h). The suppression of phosphotyrosine STAT3 levels resulted in the inhibition of STAT3 DNA binding and STAT3-mediated but not serum response element-mediated gene transcription. JSI-124 also decreased the levels of tyrosine-phosphorylated Janus kinase (JAK) but not those of Src. JSI-124 was highly selective for JAK/STAT3 and did not inhibit other oncogenic and tumor survival pathways such as those mediated by Akt, extracellular signal-regulated kinase 1/2, or c-Jun NH(2)-terminal kinase. Finally, JSI-124 (1 mg/kg/day) potently inhibited the growth in nude mice of A549 tumors, v-Src-transformed NIH 3T3 tumors, and the human breast carcinoma MDA-MB-468, all of which express high levels of constitutively activated STAT3, but it did not affect the growth of oncogenic Ras-transformed NIH 3T3 tumors that are STAT3 independent or of the human lung adenocarcinoma Calu-1, which has barely detectable levels of phosphotyrosine STAT3. JSI-124 also inhibited tumor growth and significantly increased survival of immunologically competent mice bearing murine melanoma with constitutively activated STAT3. These results give strong support for pharmacologically targeting the JAK/STAT3 signaling pathway for anticancer drug discovery. PMID- 12649186 TI - Substitution of the adenovirus serotype 5 knob with a serotype 3 knob enhances multiple steps in virus replication. AB - Adenovirus (Ad) serotype 5 (Ad5) continues to be the predominant vector used for cancer gene therapy. However, many tumor types are reported to be relatively refractory to Ad5 infection because of low surface expression of the native Ad5 receptor, CAR. The observation that many tumor cells are CAR deficient has necessitated the development of CAR-independent infection strategies, including the introduction of heterologous ligand sequences into the virus fiber gene and immunological or chemical modifications of the capsid proteins. Alternatively, native Ad5 tropism can be modified by substituting the knob region from other Ad serotypes such as Ad type 3 (Ad3) into the Ad5 knob region. To date, the effect(s) of tropism modification on the replication and oncolytic capacity of these chimeric Ad vectors has not been fully evaluated. To address this issue, Ad5 vectors and isogenically matched chimeric vectors with Ad3 tropism (Ad5/3) were compared in this study. Various parameters of virus infection were compared, including binding, nuclear translocation, E1A transcription, transgene expression, de novo virus production, and oncolysis. Overall, the chimeric Ad5/3 virus was progressively more efficient at each step of the replication cycle compared with its Ad5 counterpart. The higher replication efficiency of the chimeric Ad5/3 vector translated into improved therapeutic efficacy in a murine in vivo tumor rejection model. These findings suggest that in addition to the initial target cell interaction, multiple mechanisms contribute to the enhanced replication of the chimeric Ad5/3 vector. Furthermore, the data demonstrate that alternative Ad serotype receptors can be used to improve infection and subsequent oncolytic replication, which is particularly relevant in gene therapy applications for tumors that are inefficiently infected with Ad5. PMID- 12649188 TI - Development and preclinical evaluation of a Bacillus Calmette-Guerin-MUC1-based novel breast cancer vaccine. AB - Due to the high incidence of breast cancer and associated mortality rate,the development of an effective vaccine may be beneficial for the prevention or adjuvant treatment of this malignancy. We have constructed a novel breast cancer vaccine, Bacillus Calmette-Guerin (BCG)-hIL2MUC1, that consists of BCG and expresses a truncated form of MUC1 and human interleukin (IL)-2. In vitro analysis of the BCG-hIL2MUC1 construct confirmed coexpression of MUC1 and human IL-2. The ability of BCG-hIL2MUC1 to inhibit breast cancer growth was evaluated in hu-PBL-SCID mice (severe combined immunodeficient mice reconstituted with 50 x 10(6) human peripheral blood lymphocytes) that received three biweekly injections of BCG-hIL2MUC1 (0.5 colony-forming unit). Control animals received PBS, MUC1 peptide (100 microg), or empty vector BCG-261 (0.5 colony-forming unit) vaccination. After immunization, hu-PBL-SCID mice (n = 8 in each group) were xenografted with 4 x 10(6) ZR75-1 human breast cancer cells. Whereas mice receiving the control vaccines developed a tumor, only 87% of BCG-hIL2MUC1 immunized animals developed a palpable tumor with a slower rate of tumor growth (P < 0.001). Histological analysis of the primary tumors in BCG-hIL2MUC1 immunized animals revealed areas of reduced MUC1 expression. CD8-positive human lymphocytes were detected only in tumors grown in BCG-hIL2MUC1-immunized animals. These results imply a critical role of coexpressed IL-2 and MUC1 in eliciting tumor-specific immune response. To our knowledge, this is the first report of BCG engineered to express a tumor-associated antigen. Our results suggest that BCG hIL2MUC1 immunization inhibited breast cancer growth in hu-PBL-SCID mice. Therefore, BCG-hIL2MUC1 may be a promising candidate as a breast cancer vaccine. PMID- 12649190 TI - The role of manganese superoxide dismutase in the growth of pancreatic adenocarcinoma. AB - Chronic pancreatitis, K-ras oncogene mutations, and the subsequent generation of reactive oxygen species (ROS) appear to be linked to pancreatic cancer. ROS have also been suggested to be mitogenic and capable of stimulating cell proliferation. Cells contain antioxidant enzymes to regulate steady state levels of ROS produced by products of metabolism. The aims of our study were to determine antioxidant enzyme activity in pancreatic cancer cells and correlate enzyme activity with tumor growth, as well as determine whether tumor cell growth could be altered with antioxidant gene transfection. Western blots, enzyme activity, and enzyme activity gels were performed for manganese superoxide dismutase (MnSOD), copper/zinc, catalase, and glutathione peroxidase in normal human pancreas and in the human pancreatic cancer cell lines BxPC-3, Capan-1, MIA PaCa-2, and AsPC-1. Cell population doubling times were determined and correlated with antioxidant enzyme activity. MnSOD was overexpressed in MIA PaCa-2 using an adenoviral vector, and the effect on cell growth was determined. The cell pancreatic cancer lines BxPC-3, MIA PaCa-2, and AsPC-1 had decreased levels of MnSOD immunoreactive protein as well as activity and decreases in MnSOD levels correlated well with increased rates of tumor cell proliferation as determined by cell doubling time. No correlation could be found between cell growth and levels of copper/zinc superoxide dismutase, catalase, or glutathione peroxidase. Enforced expression of MnSOD by adenovirus transfection in the rapid growing cell line MIA PaCa-2 increased MnSOD immunoreactivity and MnSOD activity and decreased growth rate. Overexpression of MnSOD may be effective in growth suppression of pancreatic cancer. PMID- 12649189 TI - Theoretical analysis of antibody targeting of tumor spheroids: importance of dosage for penetration, and affinity for retention. AB - The interplay among antibody/antigen binding kinetics, antibody diffusion, and antigen metabolic turnover together determines the depth of penetration of antitumor antibodies into prevascular tumor spheroid cell clumps. A sharp boundary between an outer shell of bound high-affinity antibody and an inner antibody-free core has been previously observed and mathematically modeled and was termed the "binding site barrier." We show here that this process is well described by a simplified shrinking core model wherein binding equilibration is much more rapid than diffusion. This analysis provides the following experimentally testable predictions: (a) the binding site barrier is a moving boundary whose velocity is proportional to the time integral of antibody concentration at the spheroid surface (i.e. plasma antibody AUC); (b) the velocity of this moving boundary is independent of binding affinity, if the affinity is sufficiently high to strongly favor antibody/antigen complex formation at prevailing antibody concentrations; and (c) maximum tumor retention is achieved when the antibody/antigen dissociation rate approaches the rate of antigen metabolic turnover. The consistency of these predictions with published experimental results is demonstrated. The shrinking core model provides a simple analytic relationship predicting the effects of altered antibody pharmacokinetics, antibody molecular weight, antigen turnover rate, antigen expression level, and micrometastasis size on antibody penetration and retention. For example, a formula is provided for predicting the bolus dose necessary to accomplish tumor saturation as a function of antibody and tumor properties. Furthermore, this analysis indicates certain attributes necessary for an optimal tumor targeting agent. PMID- 12649191 TI - Trimetrexate inhibits progression of the murine 32Dp210 model of chronic myeloid leukemia in animals expressing drug-resistant dihydrofolate reductase. AB - Expression of drug-resistant forms of dihydrofolate reductase (DHFR) in hematopoietic cells confers substantial resistance of animals to antifolate administration. In this study, we tested whether the chemoprotection conferred by expression of the tyrosine-22 variant DHFR could be used for more effective therapy of the 32Dp210 murine model of chronic myeloid leukemia (CML). Administration of the maximum tolerated dose of trimetrexate (TMTX) with the nucleoside transport inhibitor prodrug nitrobenzylmercaptopurine ribose-5' monophosphate (NBMPR-P) inhibited 32Dp210 tumor progression in mice engrafted with transgenic tyrosine-22 DHFR marrow and improved survival of tumor-bearing animals as long as drug administration was continued. NBMPR-P coadministration was necessary for maximal tumor inhibition, as administration of TMTX alone delayed but did not prevent tumor progression. The chemoprotection afforded by engraftment with transgenic tyrosine-22 DHFR marrow was necessary for effective chemotherapy, as normal mice lacking transgenic marrow could not tolerate the higher TMTX dose (60 mg/kg/day) administered to mice with transgenic marrow, and the decreased dose of TMTX with NBMPR-P tolerated by normal tumor-bearing animals did not inhibit tumor progression or improve animal survival. We conclude that TMTX with NBMPR-P inhibits tumor progression in the 32Dp210 model of CML in animals engrafted with drug-resistant tyrosine-22 DHFR transgenic marrow, and that based on this model the introduction of a drug-resistant DHFR gene into marrow combined with TMTX and NBMPR-P administration may provide an effective treatment for CML. PMID- 12649192 TI - Enhanced cisplatin cytotoxicity by disturbing the nucleotide excision repair pathway in ovarian cancer cell lines. AB - Ovarian cancer is the leading cause of death among women from gynecological malignancies inthe United States. Resistance to the chemotherapeutic agent cisplatin isa major limitation for the successful treatment of ovarian cancer. In an effort to overcome the cisplatin resistance problem in ovarian cancer treatment, we have sought to enhance cisplatin cytotoxicity by perturbing the nucleotide excision repair (NER) pathway. The NER pathway is responsible for repairing cisplatin bound to DNA. Expression of one of the NER components, ERCC1, is correlated with cisplatin drug resistance. Hence, we targeted ERCC1 by antisense RNA methodologies, and we show that we could sensitize a relatively sensitive A2780 cell line and also the highly resistant OVCAR10 cell line to cisplatin by expressing antisense ERCC1 RNA in them as measured with 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. The A2780 cell lines expressing antisense ERCC1 had 1.9-8.1-fold enhancements in cisplatin sensitivity. The OVCAR10 antisense ERCC1 cell lines had IC(50) values ranging from 2.28 microM to 2.7 microM cisplatin as compared with 9.52 micro M for control OVCAR10 cells. The OVCAR10 antisense ERCC1 cells also show reduced DNA damage repair capacity as assessed by host cell reactivation. Furthermore, immunocompromised mice transplanted with the antisense cell lines survived longer than the mice bearing control cells after response to cisplatin treatment. These data suggest that it is possible to substantially enhance the cisplatin cytotoxicity by disturbing the NER pathway in cisplatin-resistant cell lines and to enhance the survival capacity of mice in an ovarian cancer xenograft model. PMID- 12649193 TI - Validation and analysis of a mathematical model of a replication-competent oncolytic virus for cancer treatment: implications for virus design and delivery. AB - Motivated by the rapid expansion in the development of replication-competent viral agents for the treatment of solid tumors, we formulated and analyzed a three-dimensional mathematical model of a tumor that is infected by a replication competent virus. We initially considered three patterns of intratumoral injection in which a fixed fraction of cells are initially infected with the virus throughout (a) the entire tumor, (b) the tumor core, and (c) the tumor rim, respectively. For each injection pattern, an approximate analysis of the model provides a simple and accurate condition for whether the virus will eradicate the tumor. The model was then generalized to incorporate nutrient-limited necrosis and an innate immune response against virus-infected tumor cells. Recent preclinical and clinical data were used to validate the model and estimate key parameter values. Our analysis has the following implications: even in the absence of an immune response, tumor eradication requires widespread distribution of the virus within the tumor at the time of infection; core or rim injections alone may result in tumor escape, particularly in a well-vascularized tumor; the more rapidly a virus lyses infected cells the more effective it will be at controlling the tumor; and the innate immune response to the virus can potentially prevent the virus from controlling the tumor, even with repeat injections. Therefore, in addition to diffuse intratumoral infection, tumor eradication by oncolytic adenovirus will probably require potent suppression of innate immune clearance mechanisms (e.g., by replacement of adenovirus E3 genes), combinations with traditional (chemotherapy, radiotherapy) treatments, and/or concomitant therapeutic gene expression with resultant bystander effects. PMID- 12649194 TI - Carnosic acid potentiates the antioxidant and prodifferentiation effects of 1alpha,25-dihydroxyvitamin D3 in leukemia cells but does not promote elevation of basal levels of intracellular calcium. AB - Differentiation therapy of cancer remains an only partially attained goal. Agents currently under active investigation include derivatives of vitamin D, modeled on its physiological hormone form, 1alpha,25-dihydroxyvitamin D(3) (1,25D(3)), but the calcemic effects of these compounds preclude their use in the clinic. An approach that may obviate this problem is to combine 1,25D(3) or its derivatives with other agents that increase the antineoplastic effects of low, nontoxic concentrations of vitamin D compounds. We have recently used the plant-derived polyphenolic antioxidant, carnosic acid (CA), to demonstrate an increase in the differentiating action of 1,25D(3) on human leukemia cells under these conditions (M. Danilenko et al., JNCI, 93: 1224-1233, 2001). We now show that treatment of HL60-G cells with either CA or 1,25D(3) alone resulted in a decrease in the intracellular levels of reactive oxygen species. Furthermore, the combination of 10 micro M CA and a low concentration of 1,25D(3) (1 nM) produced an enhanced antioxidant effect, which correlated with the potentiation of monocytic differentiation. Other plant antioxidants tested (curcumin, silibinin, and the organoselenium antioxidant ebselen) also potentiated differentiation induced by 1,25D(3), although alone, they had only minor differentiating effects. Differentiation induced by CA/1,25D(3) combinations was associated with increased intracellular glutathione content, whereas buthionine sulfoxime decreased both differentiation and the cellular glutathione content. This combination also enhanced the activation of the Raf-mitogen-activated protein/extracellular signal regulated kinase kinase-extracellular signal-regulated kinase mitogen-activated protein kinase module and increased the binding of the activator protein-1 (AP-1) transcription factor to its cognate DNA element in the promoter regions of vitamin D receptor gene, suggesting that the mechanism of potentiation is at least in part attributable to induction and activation of components of this mitogen-activated protein kinase pathway. Cell treatment with a high concentration of 1,25D(3) (100 nM) resulted in a substantial elevation of basal intracellular calcium concentration. In contrast, importantly for an eventual clinical application of these studies, the potentiating action of CA on differentiation induced by a low concentration of 1,25D(3) (1 nM) was not accompanied by an elevation of basal intracellular calcium concentration. These findings suggest that combinations of CA with derivatives of vitamin D should be evaluated for use in differentiation therapy of myeloid leukemias. PMID- 12649195 TI - Radioiodide treatment after sodium iodide symporter gene transfer is a highly effective therapy in neuroendocrine tumor cells. AB - This study evaluates the possibility of treating Bon1 and QGP pancreatic neuroendocrine tumor cells with radioactive iodide ((131)I) after stable transfection with the thyroid sodium iodide symporter (NIS). NIS expression was driven either by the strong viral cytomegalovirus promoter or by the tissue specific chromogranin A promoter. Using either approach, NIS expression was confirmed by reverse transcription-PCR and Western blotting. Uptake of radioactive iodide was increased approximately 20-fold by chromogranin A promoter driven NIS expression and approximately 50-fold by cytomegalovirus promoter driven NIS expression. Maximal uptake was reached within 15 min in QGP cells and 30 min in Bon1 cells. Effective half-life was 5 min in QGP and 30 min in Bon1 cells. No evidence of organification was detected by high-performance liquid chromatography and gel filtration chromatography. (131)I was a highly effective treatment in NIS-expressing QGP and Bon1 cells, reducing clone formation by 99.83 and 98.75%, respectively, in the in vitro clonogenic assay. In contrast, clone formation was not reduced in QGP and Bon1 cells without NIS expression after incubation with the same activity concentration of (131)I as compared with mock treated cells. Absorbed doses to QGP and Bon1 cells are up to 150 and 30 Gy, respectively. In addition, a direct cytotoxic effect of radioiodide was demonstrated in NIS-expressing Bon1 cells after (131)I incubation. In conclusion, radioiodide treatment after NIS gene transfer appears to be a promising novel approach in the therapy of neuroendocrine tumors if its highly encouraging in vitro effectiveness can be transferred to the in vivo situation. PMID- 12649196 TI - Mouse breast cancer resistance protein (Bcrp1/Abcg2) mediates etoposide resistance and transport, but etoposide oral availability is limited primarily by P-glycoprotein. AB - The breast cancer resistance protein [BCRP (BCRP/ABCG2)] has not previously been directly identified as a source of resistance to epipodophyllotoxins.However, when P-glycoprotein (P-gp)- and Mrp1-deficient mouse fibroblast and kidney cell lines were selected for resistance to etoposide, amplification and overexpression of Bcrp1 emerged as the dominant resistance mechanism in five of five cases. Resistance was accompanied by reduced intracellular etoposide accumulation. Bcrp1 sequence in all of the resistant lines was wild-type in the region spanning the R482 mutation hot spot known to alter the substrate specificity of mouse Bcrp1 (mouse cognate of BCRP) and human BCRP. Transduced wild-type Bcrp1 cDNA mediated resistance to etoposide and teniposide in fibroblast lines and trans-epithelial etoposide transport in polarized Madin-Darby canine kidney II cells. Bcrp1 mediated etoposide resistance was reversed by two structurally different BCRP/Bcrp1 inhibitors, GF120918 and Ko143. BCRP/Bcrp1 (inhibition) might thus impact on the antitumor activity and pharmacokinetics of epipodophyllotoxins. However, treatment of P-gp-deficient mice with GF120918 did not improve etoposide oral uptake, suggesting that Bcrp1 activity is not a major limiting factor in this process. In contrast, use of GF120918 to inhibit P-gp in wild-type mice increased the plasma levels of etoposide after oral administration 4-5-fold. It may thus be worthwhile to test inhibition of P-gp in humans to improve the oral availability of etoposide. PMID- 12649197 TI - Active specific immunotherapy against occult brain metastasis. AB - We determined whether lyophilized High Five (H5) insect cells engineered to produce IFN-beta (H5BVIFN-beta) could induce systemic immunity against occult brain metastases. C3H/HeN mice were injected s.c. with syngeneic UV-2237M fibrosarcoma or K-1735M2 melanoma cells. Intralesional injection of 2 x 10(6) lyophilized H5BVIFN-beta cells produced complete regression of the s.c. tumors. Six weeks later, UV-2237M fibrosarcoma cells or K-1735M2 melanoma cells were injected into the internal carotid artery of naive or treated mice. UV-2237M brain metastases developed in naive mice or mice cured of K-1735M2 tumors but not in mice cured of UV-2237M tumors. Similarly, K-1735M2 brain metastases developed in naive mice or mice cured of UV-2237M fibrosarcomas but not in mice cured of K 1735M2 melanoma. In the next set of studies, mice were injected s.c. with UV 2237M fibrosarcoma cells. On day 7, UV-2237M fibrosarcoma cells or K-1735M2 cells were implanted into the internal carotid artery, and on day 10, the s.c. tumors were injected with lyophilized H5BVIFN-beta. Both the s.c. tumors and the occult brain metastases produced from carotid injections were eradicated in a tumor specific manner. The regression of the brain metastases was abrogated by depletion of CD4(+) or CD8(+) cells from immunized mice. These results demonstrate that specific systemic immunity can be induced by lyophilized H5BVIFN beta and that the resultant immune response can eliminate established brain metastasis. PMID- 12649198 TI - Fusion transcripts involving HMGA2 are not a common molecular mechanism in uterine leiomyomata with rearrangements in 12q15. AB - Uterine leiomyomata are one of several benign tumors characterized by frequent chromosomal rearrangement involving 12q15. The 12q15 rearrangement in leiomyomata typically is manifested as t(12;14)(q15;q23-24), which has been hypothesized to create pathobiologically significant fusion transcripts derived from HMGA2 and RAD51L1. To explore further this hypothesis, we mapped chromosomal breakpoints in 38 uterine leiomyomata with rearrangements involving 12q15 using fluorescence in situ hybridization. Most tumors (n = 26) harbored der(14)t(12;14)(q15;q23-24), whereas chromosomes 1, 5, 8, and 10 were involved in rearrangements with 12q15 in six myomas. An additional six cases had more complex rearrangements, including breakpoints other than 12q15 or 14q23-24, inversions of chromosome 12, insertions of 12q15 into chromosome 14, or additional translocation partners. Breakpoints were mapped either 5' (centromeric) or 3' (telomeric) in the HMGA2 locus in 24 and nine cases, respectively; one tumor was a mosaic of cells with either 5' or 3' breakpoints. Breakpoints flanking the gene in both 5' and 3' regions were found in six cases. Analysis of one tumor by 3' rapid amplification of cDNA ends showed altered transcripts in which either exons 1-3 of HMGA2 were aberrantly spliced to cryptic sites in chromosome 12 or transcripts encompassing the full coding sequence of HMGA2 through a portion of the 3' untranslated region were fused to sequence from chromosome 14. A panel of 10 uterine leiomyomata with t(12;14) was specifically tested for fusion transcripts. RAD51L1-HMGA2 transcripts were not detected. HMGA2-RAD51L1 transcripts, however, were detected in four tumors; two of these tumors had uncommon rearrangements in the 3' region of HMGA2 and two had 5' rearrangements. Although the mechanism of fusion transcripts derived from tumors with 5' breakpoints is unclear, these findings indicate that formation of a fusion transcript is not the principle pathobiological mechanism in uterine leiomyomata. The pattern of rearrangements suggests dysregulated expression of HMGA2, most often by translocation of chromosome 14 sequence 5' to this gene. PMID- 12649199 TI - Requirement of RhoA activity for increased nuclear factor kappaB activity and PC 3 human prostate cancer cell invasion. AB - To determine the molecular mechanisms of aggressive prostate cancer behavior, we studied RhoGTPases in high and low invasive variants of PC-3 prostate cancer cells. Prior studies with these cells revealed that elevated nuclear factor kappaB (NF-kappaB) expression and activity were necessary for the highly invasive phenotype. In the current study, increased RhoA expression was found in the PC-3 highly invasive cells as compared with the PC-3 low invasive cells through cDNA array and Western blot analyses. Similarly, RhoA activity, as measured by the Rhotekin binding assay, was elevated in the PC-3 highly invasive cells. Transfection of these highly invasive cells with dominant negative RhoA N19 or treatment with 1.0 micro g/ml RhoA inhibitor C3 exoenzyme demonstrated that RhoA activity was necessary for both NF-kappaB activity and cellular invasion of a Matrigel reconstituted basement membrane. Furthermore, stable transfection of the PC-3 highly invasive cells with constitutively active RhoA Q63L resulted in activation of NF-kappaB activity and Matrigel invasion, effects reversed by treatment of the cells with C3 exoenzyme. RhoA was also shown to act through the motility component of the invasion process. RhoA activity was therefore both necessary and sufficient for the elevated NF-kappaB, invasion, and motility activities of the PC-3 highly invasive cells. These findings suggest molecular targets to control cancer cell invasion and aid in the development of definitive tools for predicting the invasive and metastatic potential of cancer cells. PMID- 12649200 TI - Selective Akt inactivation and tumor necrosis actor-related apoptosis-inducing ligand sensitization of renal cancer cells by low concentrations of paclitaxel. AB - Recent studies demonstrated that the resistance of cancer cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) could be reversed by various chemotherapeutic agents. In the present study, we investigated the role of Akt in the apoptosis resistance to TRAIL and chemotherapeutic agents-induced TRAIL sensitization in human renal cell carcinoma cells. Apoptosis assays and Western blot analyses revealed that apoptosis resistance to TRAIL correlates well with the level of Akt phosphorylation at Ser 473 rather than protein expression levels of TRAIL receptors, DR4 and DR5. The apoptosis sensitivity to TRAIL in TRAIL resistant SKRC-49 and TRAIL-sensitive Caki-1 cells was altered by modulation of Akt activity, which increased the protein expression of cellular FADD-like IL 1beta-converting enzyme-like inhibitory protein (cFLIP). Paclitaxel (5 and 100 nM) and cisplatin (10 microM) but not etoposide (1 and 10 microM) promoted TRAIL induced apoptosis in SKRC-49 cells, which was not mediated by increased TRAIL receptor expression but by chemotherapeutic agents-induced Akt inactivation through ceramide formation derived from sphingomyelin hydrolysis. Of note, the low concentration (5 nM) of paclitaxel promoted ceramide formation and TRAIL induced apoptosis predominantly in SKRC-49 cells but not in the normal renal proximal tubular epithelial cells. Our results may provide a novel therapeutic modality for selective killing of renal cell carcinoma with minimal toxicity on normal renal cells. PMID- 12649201 TI - Synthetic triterpenoids enhance transforming growth factor beta/Smad signaling. AB - We have studied the effects of two new synthetic triterpenoids, 2-cyano-3,12 dioxooleana-1,9-dien-28-oic acid (CDDO) and its derivative, 1-(2-cyano-3,12 dioxooleana-1,9-dien-28-oyl) imidazole (CDDO-Im), on transforming growth factor (TGF)-beta/Smad signaling. These agents, at nanomolar concentrations, increase the expression of TGF-beta-dependent genes, such as those for plasminogen activator inhibitor 1 and the type II TGF-beta receptor, and they synergize with TGF-beta in this regard. They prolong the activation of Smad2 induced by TGF-beta and markedly enhance the ability of Smad3 to activate a Smad binding element, CAGA-luciferase. In transfection assays, they reverse the inhibitory effects of Smad7. CDDO and CDDO-Im also enhance Smad signaling in the pathways of two other members of the TGF-beta superfamily, namely, activin and bone morphogenetic protein. Finally, these triterpenoids induce expression of the transcriptional coactivator p300-CBP-associated factor and synergize with TGF-beta in this regard. These are the first studies to report enhancement of Smad signaling by synthetic triterpenoids and should further their optimal use for applications in prevention or treatment of diseases in which there is aberrant function of TGF beta. PMID- 12649202 TI - Apoptotic stimuli initiate MLL-AF9 translocations that are transcribed in cells capable of division. AB - Activation of apoptosis introduces a site-specific break within intron 11 of the MLL gene. Using the CD95 apoptotic signaling pathway in human lymphoblastoid cells, the 5' fragment of MLL undergoes translocation to intron 4 of AF9 and the proleukemogenic MLL-AF9 fusion gene created is transcribed. Both the breaks in MLL and transcription of the MLL-AF9 fusion gene are suppressed in the presence of the broad spectrum caspase inhibitor, zVAD.fmk. Duplicate cells containing sequence identical MLL-AF9 fusion junctions were identified within a cell population that had recovered from apoptosis. This indicated that cells harboring a translocation initiated by apoptotic cleavage had divided. These data are consistent with a novel pathogenic role for the apoptotic program where translocations with leukemogenic potential are created within cells that have the capacity to divide. PMID- 12649203 TI - Growth and molecular profile of lung cancer cells expressing ectopic LKB1: down regulation of the phosphatidylinositol 3'-phosphate kinase/PTEN pathway. AB - Germ-line mutations in LKB1 gene cause the Peutz-Jeghers syndrome (PJS), a genetic disease with increased risk of malignancies. Recently, LKB1-inactivating mutations have been identified in one-third of sporadic lung adenocarcinomas, indicating that LKB1 gene inactivation is critical in tumors other than those of the PJS syndrome. However, the in vivo substrates of LKB1 and its role in cancer development have not been completely elucidated. Here we show that overexpression of wild-type LKB1 protein in A549 lung adenocarcinomas cells leads to cell-growth suppression. To examine changes in gene expression profiles subsequent to exogenous wild-type LKB1 in A549 cells, we used cDNA microarrays. We detected deregulation of 100 genes involved in cell proliferation, apoptosis, and cell adhesion. Strikingly, modification of the expression of well-known p53-responsive genes such as GADD45, TOP2A, and p21 suggests that growth suppression in A549 cells overexpressing LKB1 may be mediated by p53. In addition, PTEN up-regulation indicates that LKB1 could be involved in the PTEN/phosphatidylinositol-3' kinase(PI3K)/AKT molecular pathway. Thus, our results give some insights into the understanding of how LKB1 inactivation contributes to lung carcinogenesis. PMID- 12649204 TI - Female mice chimeric for expression of the simian virus 40 TAg under control of the MISIIR promoter develop epithelial ovarian cancer. AB - In women, >80% of malignant ovarian tumors are of epithelial origin. Early detection of these tumors is very challenging,and extensive i.p. dissemination is common by the time of diagnosis. The lack of adequate geneticmouse models of ovarian carcinomas significantly delays advances in early detection and treatment. We report that female transgenic mice expressing the transforming region of SV40 under control of the Mullerian inhibitory substance type II receptor gene promoter develop bilateral ovarian tumors in approximately 50% of cases. Histologically, these tumors are poorly differentiated carcinomas with occasional cysts and papillary structures present at the surface of the ovary. These tumors disseminate i.p., invade omentum, and form ascites as do human ovarian carcinomas. The epithelial origin of these tumors is supported by detection of cytokeratins 8 and 19, and the absence of alpha-inhibin, a protein characteristically expressed in normal granulosa cells and most granulosa cell tumors. Cell lines derived from the ascites exhibit the properties of epithelial ovarian cancer, such as anchorage-independent growth, tumorigenicity in immunocompromised mice, expression of epithelial cell markers, and organotropic implantation. The availability of a transgenic mouse model of disseminated ovarian carcinoma and respective cell lines should advance our understanding of this neoplasm, and serve as a useful tool for the evaluation of emerging detection and treatment strategies. PMID- 12649205 TI - Centrosome abnormalities and chromosome instability occur together in pre invasive carcinomas. AB - Centrosomes play critical roles in processes that ensure proper segregation of chromosomes and maintain the genetic stability of human cells. They contribute to mitotic spindle organization and regulate aspects of cytokinesis and cell cycle progression. We and others have shown that centrosomes are abnormal in most aggressive carcinomas. Moreover, centrosome defects have been implicated in chromosome instability and loss of cell cycle control in invasive carcinoma. Others have suggested that centrosome defects only occur late in tumorigenesis and may not contribute to early steps of tumor development. To address this issue, we examined pre-invasive human carcinoma in situ lesions for centrosome defects and chromosome instability. We found that a significant fraction of precursor lesions to some of the most common human cancers had centrosome defects, including in situ carcinomas of the uterine cervix, prostate, and female breast. Moreover, centrosome defects occurred together with mitotic spindle defects, chromosome instability, and high cytologic grade. Because most pre invasive lesions are not uniformly mutant for p53, the development of centrosome defects does not appear to require abrogation of p53 function. Our findings demonstrate that centrosome defects occur concurrently with chromosome instability and cytologic changes in the earliest identifiable step in human cancer. Our results suggest that centrosome defects may contribute to the earliest stages of cancer development through the generation of chromosome instability. This, together with ongoing structural changes in chromosomes, could accelerate accumulation of alleles carrying pro-oncogenic mutations and loss of alleles containing wild-type tumor suppressor genes and thus accelerate the genomic changes characteristic of carcinoma, the most prevalent human cancer. PMID- 12649206 TI - Hepatocyte growth factor production by neutrophils infiltrating bronchioloalveolar subtype pulmonary adenocarcinoma: role in tumor progression and death. AB - Increased numbers of tumor-infiltrating neutrophils are linked to poorer outcome in patients with adenocarcinoma of the bronchioloalveolar carcinoma (BAC) subtype. Hepatocyte growth factor (HGF) is a pleiotropic cytokine operating through activation of the proto-oncogene c-met and is a factor of poor prognosis in various cancers. Reports that neutrophils produce HGF led us to investigate their participation in the aerogenous spread of tumor cells and the prognosis of BAC, through the effect of HGF on c-met-expressing tumor cells. Immunoreactive HGF was detected in bronchoalveolar lavage fluid (BALF) supernatants from 34 of 36 patients, whereas it was undetectable in BALF from healthy controls. The HGF thus detected was locally produced, because HGF mRNA was expressed by the patients' fresh alveolar cells, and HGF protein was detected in 24-h culture supernatants. In immunocytochemical studies of BALF cytospin preparations and tumor specimens from the patients, neutrophils were always HGF-positive, whereas alveolar macrophages and tumor cells gave inconsistent results. Alveolar neutrophil-derived HGFs induced significant, concentration-dependent migration of BAC-derived tumor cells in vitro, and this effect was inhibited by anti-HGF neutralizing antibodies. Granulocyte-macrophage colony-stimulating factor and tumor necrosis factor alpha (present in the lung tumor microenvironment) provoked HGF release from neutrophil intracellular stocks, and the capacity of blood neutrophils from BAC patients to produce HGF was unaltered. Immunochemical studies of c-met expression in BALF cytospin preparations and tumor sections showed that most HGF receptor-bearing cells were tumor cells. High HGF levels in BALF supernatants were significantly associated with poorer outcome in patients with BAC and were an independent predictor of clinical outcome in multivariate analysis. Altogether, our results support the notion that BAC generates a local environment that attracts functionally normal neutrophils from peripheral blood and leads to neutrophil release of biologically active HGF on contact with HGF receptor-expressing tumor cells, thereby contributing to poorer patient outcome. PMID- 12649207 TI - Loss of c-myc repression coincides with ovarian cancer resistance to transforming growth factor beta growth arrest independent of transforming growth factor beta/Smad signaling. AB - Many epithelial carcinomas, including ovarian, are refractory to the antiproliferative effects of transforming growth factor (TGF) beta. In some cancers, TGF-beta resistance has been linked to TGF-beta receptor II (TbetaR-II) and Smad4 mutations; however, in ovarian cancer, the mechanism of resistance remains unclear. Primary ovarian epithelial cell cultures were used as a model system to determine the mechanisms of TGF-beta resistance. To simulate in vivo responses to TGF-beta, primary cultures derived from normal human ovarian surface epithelium (HOSE) and from ovarian carcinomas (CSOC) were grown on collagen I gel, the predominant matrix molecule in the ovarian tumor milieu. When treated with 5 ng/ml TGF-beta for 72 h, HOSE (n = 11) proliferation was inhibited by 20 +/- 21% on average. In contrast, CSOC (n = 10) proliferation was stimulated 5 +/- 10% in response to TGF-beta (a statistically significant difference in response when compared with HOSE; P = 0.001). To dissect the TGF-beta/Smad signaling pathway we used a quantitative RNase protection assay (RPA) for measuring mRNA levels of TGF-beta pathway components in 20 HOSE and 20 CSOC cultures. Basal mRNA levels of TGF-beta receptors I and II, downstream signaling components Smad2, 3, 4, 6, 7, and the transcriptional corepressors Ski and SnoN did not show a statistically significant difference between HOSE and CSOC, and cannot explain their differential susceptibility to TGF-beta-induced cell cycle arrest. To assess functional differences of the TGF-beta pathway in TGF-beta-sensitive HOSE and TGF-beta-resistant CSOC, we measured Smad2/4 and 3/4 complex induction after TGF-beta treatment. HOSE and CSOC showed equivalent Smad2/4 and 3/4 complex induction after TGF-beta exposure for 0, 0.5, 2, and 4 h. It has been proposed that SnoN and Ski are corepressors of the TGF-beta/Smad pathway and undergo TGF beta-induced degradation followed by reinduction of SnoN mRNA. However, our data show equivalent SnoN degradation in HOSE and CSOC, and equivalent SnoN mRNA induction after TGF-beta treatment. Surprising, TGF-beta-induced Ski degradation was not observed in HOSE or CSOC, suggesting that Ski may not function as a TGF beta/Smad corepressor in ovarian epithelial cells. These data implied that the TGF-beta/Smad pathway remains functional in CSOC, although CSOC cells are resistant to antimitogenic TGF-beta effects. CSOC resistance to TGF-beta coincided with the loss of c-myc down-regulation. These data suggest that TGF beta/Smad signaling is blocked downstream of Smad complex formation or that an alternate signaling pathway other than TGF-beta/Smad may transmit TGF-beta induced cell cycle arrest in the ovarian epithelium. PMID- 12649208 TI - The tyrphostin AG1024 accelerates the degradation of phosphorylated forms of retinoblastoma protein (pRb) and restores pRb tumor suppressive function in melanoma cells. AB - Constitutive cell surface receptor kinase signaling and persistent phosphorylation/inactivation of the retinoblastoma (pRb) family of proteins (pRb, p107 and p130, known as pocket proteins) have been implicated in conferring uncontrolled growth to melanoma cells. However, the signals linking receptor kinase activity to neutralization of pocket proteins have not yet been fully elucidated. We therefore used specific chemical inhibitors to examine pRb regulation in melanoma cells. The most efficient agent, AG1024, known as an inhibitor of insulin-like growth factor 1 receptor and insulin receptor, arrested melanoma cell growth in vitro at nanomolar concentrations within 24 h of application. AG1024 inhibited the mitogen-activated protein kinase/extracellular signal-regulated kinase pathway and restored pRb tumor suppressive function. The latter was observed by the reduction in the phosphorylated forms of pRb, p107 and p130, and the formation of growth suppressive DNA binding complexes consisting of pRb and E2F1 or E2F3. The loss of phosphorylated forms of pRb at early time points after AG1024 application was not associated with suppression of cyclin dependent kinases 2 and 4 activity but rather with proteasomal and nonproteasomal degradation. Thus, inhibition of melanoma cell proliferation by AG1024 is mediated by inhibition of mitogen-activated protein kinase/extracellular signal regulated kinase 2 signaling and activation of pRb by a mechanism involving protein degradation. PMID- 12649209 TI - E1A deregulates the centrosome cycle in a Ran GTPase-dependent manner. AB - By means of the yeast two-hybrid system, we have discovered a novel physical interaction between the adenovirus E1A oncoprotein and Ran, a small GTPase which regulates nucleocytoplasmic transport, cell cycle progression, and mitotic spindle organization. Expression of E1A elicits induction of S phase and centrosome amplification in a variety of rodent cell lines. The induction of supernumerary centrosomes requires functional RCC1, the nucleotide exchange factor for Ran and, hence, a functional Ran network. The E1A portion responsible for the interaction with Ran is the extreme NH(2)-terminal region (amino acids 1 36), which is also required for the induction of centrosome amplification. In an in vitro assay with recombinant proteins, wild-type E1A interferes with nucleotide exchange on Ran, whereas an E1A mutant, deleted from the extreme NH(2) terminal region, does not. In addition, we detected an in vitro interaction between Ran and HPV-16 E7 and SV40 large T antigen, two oncoproteins functionally related to E1A. These findings suggest a common pathway of these oncoproteins in eliciting virus-induced genomic instability. PMID- 12649210 TI - Doctors and managers: a problem without a solution? PMID- 12649211 TI - What doctors and managers can learn from each other. PMID- 12649212 TI - Doctors in chambers. PMID- 12649213 TI - Useful information about rare inherited disorders. PMID- 12649214 TI - Not so benign intracranial hypertension. PMID- 12649215 TI - WHO issues global alert on respiratory syndrome. PMID- 12649216 TI - UK travellers warned after first suspected case of new syndrome. PMID- 12649219 TI - Privatisation of water is high on Kyoto summit agenda. PMID- 12649218 TI - GMC accuses doctor of research fraud. PMID- 12649223 TI - US buyers of drugs from Canadian pharmacies face prosecution. PMID- 12649225 TI - Lawyers may seek judicial review of panel reviewing paroxetine. PMID- 12649227 TI - US Senate outlaws "partial birth abortion". PMID- 12649229 TI - UK nuclear medicine is close to collapse, experts warn. PMID- 12649230 TI - Whistleblower charges drug company with deceptive practices. PMID- 12649231 TI - US body reviews errors in emergency departments. PMID- 12649232 TI - Possible link between flu jab and Guillain-Barre syndrome under investigation. PMID- 12649233 TI - Inhaled glucocorticoids versus leukotriene receptor antagonists as single agent asthma treatment: systematic review of current evidence. AB - OBJECTIVE: To compare the safety and efficacy of anti-leukotrienes and inhaled glucocorticoids as monotherapy in people with asthma. DESIGN: Systematic review of randomised controlled trials comparing anti-leukotrienes with inhaled glucocorticoids for 28 days or more in children and adults. MAIN OUTCOME MEASURE: Rate of exacerbations that required treatment with systemic glucocorticoids. RESULTS: 13 trials (12 in adults, one in children) met the inclusion criteria; all were in people with mild and moderate asthma. Leukotriene receptor antagonists were compared with inhaled glucocorticoids at a daily dose equivalent to 400-450 microg beclometasone dipropionate. Patients treated with leukotriene receptor antagonists were 60% more likely to suffer an exacerbation requiring systemic glucocorticoids (relative risk 1.6, 95% confidence interval 1.2 to 2.2; number needed to treat 27, 13 to 81). A 130 ml greater improvement (80 ml to 170 ml) in forced expiratory volume in one second and a 19 l/min greater increase (14 l to 24 l) in morning peak expiratory flow rate were noted in favour of inhaled glucocorticoids. Differences in favour of inhaled glucocorticoids were also observed for nocturnal awakenings, use of rescue beta2 agonists, and days without symptoms. Risk of side effects was no different between groups, but leukotriene receptor antagonists were associated a 2.5-fold increase risk of withdrawals due to poor asthma control (relative risk 2.5, 1.8 to 3.5). CONCLUSIONS: Inhaled glucocorticoids doses equivalent to 400 microg/day beclometasone are more effective than leukotriene receptor antagonists in the treatment of adults with mild or moderate asthma. There is insufficient evidence to conclude on the efficacy of anti-leukotrienes in children. PMID- 12649234 TI - Central overweight and obesity in British youth aged 11-16 years: cross sectional surveys of waist circumference. AB - OBJECTIVE: To compare changes over time in waist circumference (a measure of central fatness) and body mass index (a measure of overall obesity) in British youth. DESIGN: Representative cross sectional surveys in 1977, 1987, and 1997. SETTING: Great Britain. PARTICIPANTS: Young people aged 11-16 years surveyed in 1977 (boys) and 1987 (girls) for the British Standards Institute (n=3784) and in 1997 (both sexes) for the national diet and nutrition survey (n=776). MAIN OUTCOME MEASURES: Waist circumference, expressed as a standard deviation score using the first survey as reference, and body mass index (weight(kg)/height(m)2), expressed as a standard deviation score against the British 1990 revised reference. Overweight and obesity were defined as the measurement exceeding the 91st and 98th centile, respectively. RESULTS: Waist circumference increased sharply over the period between surveys (mean increases for boys and girls, 6.9 and 6.2 cm, or 0.84 and 1.02 SD score units, P<0.0001). In centile terms, waist circumference increased more in girls than in boys. Increases in body mass index were smaller and similar by sex (means 1.5 and 1.6, or 0.47 and 0.53 SD score units, P<0.0001). Waist circumference in 1997 exceeded the 91st centile in 28% (n=110) of boys and 38% (n=147) of girls (against 9% for both sexes in 1977-87, P<0.0001), whereas 14% (n=54) and 17% (n=68), respectively, exceeded the 98th centile (3% in 1977-87, P<0.0001). The corresponding rates for body mass index in 1997 were 21% (n=80) of boys and 17% (n=67) of girls exceeding the 91st centile (8% and 6% in 1977-87) and 10% (n=39) and 8% (n=32) exceeding the 98th centile (3% and 2% in 1977-87). CONCLUSIONS: Trends in waist circumference during the past 10-20 years have greatly exceeded those in body mass index, particularly in girls, showing that body mass index is a poor proxy for central fatness. Body mass index has therefore systematically underestimated the prevalence of obesity in young people. PMID- 12649235 TI - Views of doctors and managers on the doctor-manager relationship in the NHS. PMID- 12649237 TI - Media criticism of doctors: review of UK junior doctors' concerns raised in surveys. PMID- 12649236 TI - Evidence of Plasmodium falciparum malaria resistant to atovaquone and proguanil hydrochloride: case reports. PMID- 12649238 TI - Primary care in the United States: organisation of primary care in the United States. PMID- 12649240 TI - Interactive case report: a 66 year old woman with a rash: case progression. PMID- 12649239 TI - Recent developments: management of pain. PMID- 12649242 TI - ABC of learning and teaching in medicine: Written assessment. PMID- 12649241 TI - Doxycycline induced intracranial hypertension. PMID- 12649243 TI - Trends in doctor-manager relationships. PMID- 12649244 TI - Medicine, management, and modernisation: a "danse macabre"? PMID- 12649246 TI - Improving the doctor-manager relationship. Doctors and managers: mind the gap. PMID- 12649247 TI - Improving the doctor-manager relationship. Building a successful partnership between management and clinical leadership: experience from New Zealand. PMID- 12649248 TI - Improving the doctor-manager relationship. Kaiser Permanente: a propensity for partnership. PMID- 12649250 TI - Improving the doctor-manager relationship. Clinical networks for doctors and managers. PMID- 12649249 TI - Improving the doctor-manager relationship. Doctors and managers need to speak a common language. PMID- 12649251 TI - Doctors and managers. Agreeing objectives could help doctors and managers work well together. PMID- 12649252 TI - Values and leadership. Leadership is failing to adhere to values in the NHS. PMID- 12649253 TI - The making of a disease: female sexual dysfunction. Without industry funding little new research will be possible. PMID- 12649254 TI - Company defends "campaign to 'counter' BMJ claims". PMID- 12649256 TI - Sensory stimulation in dementia. Cause of behavioural and psychological symptoms of dementia needs to be established first. PMID- 12649257 TI - Musculoskeletal medicine. PMID- 12649259 TI - Modeling development of the epicardium and coronary vasculature: in vitro veritas? PMID- 12649260 TI - From pump to molecules. PMID- 12649261 TI - PPARs of the heart: three is a crowd. PMID- 12649262 TI - Letter to the editor: regarding authorship. PMID- 12649263 TI - Myocytes die by multiple mechanisms in failing human hearts. AB - We tested the hypothesis that myocyte loss in failing human hearts occurs by different mechanisms: apoptosis, oncosis, and autophagic cell death. Explanted hearts from 19 patients with idiopathic dilated cardiomyopathy (EF< or =20%) and 7 control hearts were analyzed. Myocyte apoptosis revealed by caspase-3 activation and TUNEL staining occurred at a rate of 0.002+/-0.0005% (P<0.05 versus control) and oncosis assessed by complement 9 labeling at 0.06+/-0.001% (P<0.05). Cellular degeneration including appearance of ubiquitin containing autophagic vacuoles and nuclear disintegration was present at the ultrastructural level. Nuclear and cytosolic ubiquitin/protein accumulations occurred at 0.08+/ 0.004% (P<0.05). The ubiquitin-activating enzyme E1 and the ligase E3 were not different from control. In contrast, ubiquitin mRNA levels were 1.8-fold (P<0.02) elevated, and the conjugating enzyme E2 was 2.3-fold upregulated (P<0.005). The most important finding, however, is the 2.3-fold downregulation of the deubiquitination enzyme isopeptidase-T and the 1.5-fold reduction of the ubiquitin-fusion degradation system-1, which in conjunction with unchanged proteasomal subunit levels and proteasomal activity results in massive storage of ubiquitin/protein complexes and in autophagic cell death. A 2-fold decrease of cathepsin D might be an additional factor responsible for the accumulation of ubiquitin/protein conjugates. It is concluded that in human failing hearts apoptosis, oncosis, and autophagy act in parallel to varying degrees. A disturbed balance between a high rate of ubiquitination and inadequate degradation of ubiquitin/protein conjugates may contribute to autophagic cell death. Together, these different types of cell death play a significant role for myocyte disappearance and the development of contractile dysfunction in failing hearts. PMID- 12649264 TI - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) sequentially upregulates nitric oxide and prostanoid production in primary human endothelial cells. AB - Endothelial cells express tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptors, but the function of TRAIL in endothelial cells is not completely understood. We explored the role of TRAIL in regulation of key intracellular signal pathways in endothelial cells. The addition of TRAIL to primary human endothelial cells increased phosphorylation of endothelial nitric oxide synthase (eNOS), NOS activity, and NO synthesis. Moreover, TRAIL induced cell migration and cytoskeleton reorganization in an NO-dependent manner. TRAIL did not activate the NF-kappaB or COX-2 pathways in endothelial cells. Instead, TRAIL increased prostanoid production (PGE2=PGI2>TXA2), which was preferentially inhibited by the COX-1 inhibitor SC-560. Because NO and prostanoids play a crucial role in the state of blood vessel vasodilatation and angiogenesis, our data suggest that TRAIL might play an important role in endothelial cell function. PMID- 12649265 TI - MAP kinase kinase 6-p38 MAP kinase signaling cascade regulates cyclooxygenase-2 expression in cardiac myocytes in vitro and in vivo. AB - Cyclooxygenase-2 (COX-2) catalyzes the rate-limiting step in delayed prostaglandin biosynthesis. The purpose of this study was to evaluate the role of the MAP kinase kinase 6 (MKK6)-p38 MAPK signaling cascade in the regulation of myocardial COX-2 gene expression, in vitro and in vivo. RT-PCR analysis identified p38alpha and p38beta2 MAPK mRNA in rat cardiac myocytes. Interleukin 1beta induced the phosphorylation of p38alpha and p38beta2 MAPK in cardiomyocytes and stimulated RNA polymerase II binding to the COX-2 promoter, COX-2 transcription, COX-2 protein synthesis, and prostaglandin E2 (PGE2) release. Infecting cardiomyocytes with adenoviruses that encode mutant, phosphorylation resistant MKK6 or p38beta2 MAPK inhibited interleukin-1beta-induced p38 MAPK activation, COX-2 gene expression, and PGE2 release. Treatment with the p38alpha and p38beta2 MAPK inhibitor, SB202190, attenuated interleukin-1beta-induced COX-2 transcription and accelerated the degradation of COX-2 mRNA. Cells infected with adenoviruses encoding wild-type or constitutively activated MKK6 or p38beta2 MAPK, in the absence of interleukin-1beta, exhibited increased cellular p38 MAPK activity, COX-2 mRNA expression, and COX-2 protein synthesis, which was blocked by SB202190. In addition, elevated levels of COX-2 protein were identified in the hearts of transgenic mice with cardiac-restricted expression of wild-type or constitutively activated MKK6, in comparison with nontransgenic littermates. These results provide direct evidence that MKK6 mediated p38 MAPK activation is necessary for interleukin-1beta-induced cardiac myocyte COX-2 gene expression and PGE2 biosynthesis in vitro and is sufficient for COX-2 gene expression by cardiac myocytes in vitro and in vivo. PMID- 12649266 TI - Ineffectiveness of histone deacetylase inhibitors to induce apoptosis involves the transcriptional activation of NF-kappa B through the Akt pathway. AB - Histone deacetylase (HDAC) inhibitors are emerging as a new class of anticancer agents for the treatment of solid and hematological malignancies. Although HDAC inhibitors induce cell death through an apoptotic process, little is known about the molecular events that control their effectiveness. In this study, we demonstrate that HDAC inhibitors are limited in their ability to induce apoptosis in non-small cell lung cancer (NSCLC) cell lines despite their ability to effectively inhibit deacetylase activity. Because the anti-apoptotic transcription factor NF-kappa B has been shown to be under the control of HDAC mediated repression, we analyzed whether HDAC inhibitors activated NF-kappa B in NSCLC cells. HDAC inhibitors effectively stimulated endogenous NF-kappa B dependent gene expression by up-regulating IL-8, Bcl-XL, and MMP-9 transcripts. The ability of HDAC inhibitors to increase NF-kappa B transcriptional activity was not associated with signaling events that stimulated nuclear translocation, but rather modulated the transactivation potential of the RelA/p65 subunit of NF kappa B. The inhibition of HDAC activity was associated with the recruitment of the p300 transcriptional co-activator to chromatin in an Akt-dependent manner. Moreover, Akt directly phosphorylated p300 in vitro and was required for stimulating the transactivation potential of the co-activator following the addition of HDAC inhibitors. Selective inhibition of either the phosphoinositide 3-kinase/Akt pathway, or NF-kappa B itself blocked the ability of HDAC inhibitors to activate NF-kappa B and dramatically sensitized NSCLC cells to apoptosis following of the addition of HDAC inhibitors. Our study indicates that the ineffectiveness of HDAC inhibitors to induce apoptosis in NSCLC cancer cells is associated with the ability of these molecules to stimulate NF-kappa B-dependent transcription and cell survival. PMID- 12649267 TI - Lens epithelium-derived growth factor relieves transforming growth factor-beta1 induced transcription repression of heat shock proteins in human lens epithelial cells. AB - Lens epithelium-cell derived growth factor (LEDGF) is a transcriptional activator. It protects the cells by binding to cis-stress response ((A/T)GGGG(T/A)), and heat shock (HSE; nGAAn) elements in the stress genes and activating their transcription. Transforming growth factor-beta (TGF-beta) has been implicated in the control of tissue homeostasis, terminal differentiation, and apoptosis. Here we provide evidence that TGF-beta1 down-regulates LEDGF expression and diminishes its affinity for DNA during TGF-beta1-induced phenotypic changes and apoptosis in human lens epithelial cells. Surprisingly, TGF-beta1 treatment for 48 h markedly decreased the LEDGF, Hsp27, and alphaB crystallin promoter activities with the decrease of abundance of LEDGF mRNA and protein. Deletion mutants of the LEDGF promoter showed that one TGF-beta1 inhibitory element (TIE) like sequence nnnTTGGnnn (-444 to -433) contributed to this negative regulation. Mutation of TIE (TTGG to TATT) abolished the down regulation of the LEDGF promoter. Gel mobility and supershift assays showed that LEDGF in the nuclear extracts of TGF-beta1-treated human lens epithelial cells did not bind to stress-response elements and HSE. The TGF-beta1-induced down regulation of LEDGF, Hsp27, and alphaB-crystallin promoters activity was reversed by cotransfection with a plasmid expressing LEDGF. Because overexpression of LEDGF was able to relieve TGF-beta1 and/or stress-induced changes, it would be a candidate molecule to postpone age-related degenerating disorders. PMID- 12649268 TI - Two trans-sialidase forms with different sialic acid transfer and sialidase activities from Trypanosoma congolense. AB - Trypanosomes express an enzyme called trans-sialidase (TS), which enables the parasites to transfer sialic acids from the environment onto trypanosomal surface molecules. Here we describe the purification and characterization of two TS forms from the African trypanosome Trypanosoma congolense. The purification of the two TS forms using a combination of anion exchange chromatography, isoelectric focusing, gel filtration, and subsequently, antibody affinity chromatography resulted, in both cases, in the isolation of a 90-kDa monomer on SDS-PAGE, which was identified as trans-sialidase using micro-sequencing. Monoclonal antibody 7/23, which bound and partially inhibited TS activity, was found in both cases to bind to a 90-kDa protein. Both TS forms possessed sialidase and transfer activity, but markedly differed in their activity ratios. The TS form with a high transfer-to-sialidase activity ratio, referred to as TS-form 1, possessed a pI of pH 4-5 and a molecular mass of 350-600 kDa. In contrast, the form with a low transfer-to-sialidase activity ratio, referred to as TS-form 2, exhibited a pI of pH 5-6.5 and a molecular mass of 130-180 kDa. Both TS forms were not significantly inhibited by known sialidase inhibitors and revealed no significant differences in donor and acceptor substrate specificities; however, TS-form 1 utilized various acceptor substrates with a higher catalytic efficiency. Interestingly, glutamic acid-alanine-rich protein, the surface glycoprotein, was co-purified with TS-form 1 suggesting an association between both proteins. PMID- 12649269 TI - Stimulation of cellular signaling and G protein subunit dissociation by G protein betagamma subunit-binding peptides. AB - We previously developed peptides that bind to G protein betagamma subunits and selectively block interactions between betagamma subunits and a subset of effectors in vitro (Scott, J. K., Huang, S. F., Gangadhar, B. P., Samoriski, G. M., Clapp, P., Gross, R. A., Taussig, R., and Smrcka, A. V. (2001) EMBO J. 20, 767-776). Here, we created cell-permeating versions of some of these peptides by N-terminal modification with either myristate or the cell permeation sequence from human immunodeficiency virus TAT protein. The myristoylated betagamma binding peptide (mSIRK) applied to primary rat arterial smooth muscle cells caused rapid activation of extracellular signal-regulated kinase 1/2 in the absence of an agonist. This activation did not occur if the peptide lacked a myristate at the N terminus, if the peptide had a single point mutation to eliminate betagamma subunit binding, or if the cells stably expressed the C terminus of betaARK1. A human immunodeficiency virus TAT-modified peptide (TAT SIRK) and a myristoylated version of a second peptide (mSCAR) that binds to the same site on betagamma subunits as mSIRK, also caused extracellular signal regulated kinase activation. mSIRK also stimulated Jun N-terminal kinase phosphorylation, p38 mitogen-activated protein kinase phosphorylation, and phospholipase C activity and caused Ca2+ release from internal stores. When tested with purified G protein subunits in vitro, SIRK promoted alpha subunit dissociation from betagamma subunits without stimulating nucleotide exchange. These data suggest a novel mechanism by which selective betagamma-binding peptides can release G protein betagamma subunits from heterotrimers to stimulate G protein pathways in cells. PMID- 12649270 TI - Crystal structure of Enterococcus faecalis SlyA-like transcriptional factor. AB - The crystal structure of a SlyA transcriptional regulator at 1.6 A resolution is presented, and structural relationships between members of the MarR/SlyA family are discussed. The SlyA family, which includes SlyA, Rap, Hor, and RovA proteins, is widely distributed in bacterial and archaeal genomes. Current evidence suggests that SlyA-like factors act as repressors, activators, and modulators of gene transcription. These proteins have been shown to up-regulate the expression of molecular chaperones, acid-resistance proteins, and cytolysin, and down regulate several biosynthetic enzymes. The structure of SlyA from Enterococcus faecalis, determined as a part of an ongoing structural genomics initiative (www.mcsg.anl.gov), revealed the same winged helix DNA-binding motif that was recently found in the MarR repressor from Escherichia coli and the MexR repressor from Pseudomonas aeruginosa, a sequence homologue of MarR. Phylogenetic analysis of the MarR/SlyA family suggests that Sly is placed between the SlyA and MarR subfamilies and shows significant sequence similarity to members of both subfamilies. PMID- 12649271 TI - Ceramide stabilizes beta-site amyloid precursor protein-cleaving enzyme 1 and promotes amyloid beta-peptide biogenesis. AB - The lipid second messenger ceramide regulates several biochemical events that occur during aging. In addition, its level is highly elevated in the amyloid burdened brains of Alzheimer's disease patients. Here, we analyzed the impact of aberrant ceramide levels on amyloid beta-peptide (Abeta) generation by using a cell-permeable analog of ceramide, C6-ceramide, and several biochemical inhibitors of the sphingomyelin/glycosphingolipid biosynthetic pathway. We found that C6-ceramide increased the biogenesis of Abeta by affecting beta-but not gamma-cleavage of the amyloid precursor protein. Similarly to C6-ceramide, increased levels of endogenous ceramide induced by neutral sphingomyelinase treatment also promoted the biogenesis of Abeta. Conversely, fumonisin B1, which inhibits the biosynthesis of endogenous ceramide, reduced Abeta production. Exogenous C6-ceramide restored both intracellular ceramide levels and Abeta generation in fumonisin B1-treated cells. These events were specific for amyloid precursor protein and were not associated with apoptotic cell death. Pulse-chase and time-course degradation experiments showed that ceramide post-translationally stabilizes the beta-secretase BACE1. Taken together, these data indicate that the lipid second messenger ceramide, which is elevated in the brains of Alzheimer's disease patients, increases the half-life of BACE1 and thereby promotes Abeta biogenesis. PMID- 12649272 TI - An alternative mechanism of bicarbonate-mediated peroxidation by copper-zinc superoxide dismutase: rates enhanced via proposed enzyme-associated peroxycarbonate intermediate. AB - Hydrogen peroxide can interact with the active site of copper-zinc superoxide dismutase (SOD1) to generate a powerful oxidant. This oxidant can either damage amino acid residues at the active site, inactivating the enzyme (the self oxidative pathway), or oxidize substrates exogenous to the active site, preventing inactivation (the external oxidative pathway). It is well established that the presence of bicarbonate anion dramatically enhances the rate of oxidation of exogenous substrates. Here, we show that bicarbonate also substantially enhances the rate of self-inactivation of human wild type SOD1. Together, these observations suggest that the strong oxidant formed by hydrogen peroxide and SOD1 in the presence of bicarbonate arises from a pathway mechanistically distinct from that producing the oxidant in its absence. Self inactivation rates are further enhanced in a mutant SOD1 protein (L38V) linked to the fatal neurodegenerative disorder, familial amyotrophic lateral sclerosis. The 1.4 A resolution crystal structure of pathogenic SOD1 mutant D125H reveals the mode of oxyanion binding in the active site channel and implies that phosphate anion attenuates the bicarbonate effect by competing for binding to this site. The orientation of the enzyme-associated oxyanion suggests that both the self oxidative and external oxidative pathways can proceed through an enzyme associated peroxycarbonate intermediate. PMID- 12649273 TI - The UDP-glucose:glycoprotein glucosyltransferase is organized in at least two tightly bound domains from yeast to mammals. AB - The endoplasmic reticulum UDP-Glc:glycoprotein glucosyltransferase (GT) exclusively glucosylates nonnative glycoprotein conformers. GT sequence analysis suggests that it is composed of at least two domains: the N-terminal domain, which composes 80% of the molecule, has no significant similarity to other known proteins and was proposed to be involved in the recognition of non-native conformers and the C-terminal or catalytic domain, which displays a similar size and significant similarity to members of glycosyltransferase family 8. Here, we show that N- and C-terminal domains from Rattus norvegicus and Schizosaccharomyces pombe GTs remained tightly but not covalently bound upon a mild proteolytic treatment and could not be separated without loss of enzymatic activity. The notion of a two-domain protein was reinforced by the synthesis of an active enzyme upon transfection of S. pombe GT null mutants with two expression vectors, each of them encoding one of both domains. Transfection with the C-terminal domain-encoding vector alone yielded an inactive, rapidly degraded protein, thus indicating that the N-terminal domain is required for proper folding of the C-terminal catalytic portion. If, indeed, the N-terminal domain is, as proposed, also involved in glycoprotein conformation recognition, the tight association between N- and C-terminal domains may explain why only N glycans in close proximity to protein structural perturbations are glucosylated by the enzyme. Although S. pombe and Drosophila melanogaster GT N-terminal domains display an extremely poor similarity (16.3%), chimeras containing either yeast N-terminal and fly C-terminal domains or the inverse construction were enzymatically and functionally active in vivo, thus indicating that the N terminal domains of both GTs shared three-dimensional features. PMID- 12649274 TI - Tpn1p, the plasma membrane vitamin B6 transporter of Saccharomyces cerevisiae. AB - Pyridoxine (PN) is a metabolic precursor of pyridoxal phosphate that functions as a cofactor of many enzymes in amino acid metabolism. PN, pyridoxal, and pyridoxamine are collectively referred to as vitamin B6, and mammalian organisms depend on its uptake from the diet. In addition to the ability to use extracellular vitamin B6, most unicellular organisms are also capable of synthesizing PN to generate pyridoxal phosphate. Here, we report the isolation of Saccharomyces cerevisiae mutants that have lost the ability to transport PN across the plasma membrane. We used these mutants to isolate TPN1, the first known gene encoding a transport protein for vitamin B6. Tpn1p is a member of the purine-cytosine permease family within the major facilitator superfamily. The protein functions as a proton symporter, localizes to the plasma membrane, and has high affinity for PN. TPN1 mutants lost the ability to utilize extracellular PN, pyridoxal, and pyridoxamine, showing that there is no other transporter for vitamin B6 encoded in the genome. Amino acid substitutions that led to a loss of Tpn1p function localized to transmembrane domain 4 within the 12-transmembrane domain protein. Moreover, expression of TPN1 was regulated and increased with decreasing concentrations of vitamin B6 in the medium. We also provide evidence that of the highly conserved SNZ and SNO genes in S. cerevisiae, only the protein encoded by SNZ1 is required for vitamin B6 biosynthesis. PMID- 12649275 TI - Interaction of guanylyl cyclase C with SH3 domain of Src tyrosine kinase. Yet another mechanism for desensitization. AB - Protein-protein interactions mediated by the Src homology 3 (SH3) domain have been implicated in the regulation of receptor functions for subcellular localization of proteins and the reorganization of cytoskeleton. The experiments described in this article begin to identify the interaction of the SH3 domain of Src tyrosine kinase with the guanylyl cyclase C receptor after activation with Escherichia coli heat-stable enterotoxin (ST). Only one of two post translationally modified forms of guanylyl cyclase C from T84 colonic carcinoma cells bind to GST-SH3 fusion protein of Src and Hck tyrosine kinases. Interestingly, the GST-Src-SH3 fusion protein showed 2-fold more affinity to native guanylyl cyclase C in solution than the GST-Hck-SH3 fusion protein. The affinity of the GST-Src-SH3 fusion protein to guanylyl cyclase C increased on desensitization of receptor in vivo. An in vitro cyclase assay in the presence of GST-Src-SH3 fusion protein indicated inhibition of the catalytic activity of guanylyl cyclase C. The catalytic domain recombinant protein (GST-GCD) of guanylyl cyclase C could pull-down a 60-kDa protein that reacted with Src tyrosine antibody and also showed autophosphorylation. These data suggest that SH3 domain-mediated protein-protein interaction with the catalytic domain of guanylyl cyclase C inhibited the cyclase activity and that such an interaction, possibly mediated by Src tyrosine kinase or additional proteins, might be pivotal for the desensitization phenomenon of the guanylyl cyclase C receptor. PMID- 12649276 TI - Detection of an intermediate during unfolding of bacterial cell division protein FtsZ: loss of functional properties precedes the global unfolding of FtsZ. AB - Using environment-sensitive fluorescence of 1-anilinonaphthalene-8-sulfonic acid, polarization of fluorescein 5'-isothiocyanate-labeled FtsZ, and far-UV circular dichroism spectroscopy, the chemical unfolding of FtsZ was found to proceed through two steps. The first step of the urea-induced unfolding produced an intermediate, which then unfolded at higher concentrations of urea. The intermediate state contains native-like secondary structure and much less tertiary structure compared with the native state. It is distinct from the native state as well as from the unfolded state. Similar to urea-induced unfolding of FtsZ, thermal unfolding of FtsZ also occurs in two steps. The midpoints for the first and second thermal unfolding transitions were found to be 38 +/- 4 and 77 +/- 5 degrees C, respectively. Further, the functional properties of FtsZ are extremely sensitive to urea, guanidium chloride, and sodium dodecyl sulfate. For example, 50% inhibition of the FtsZ assembly and GTP hydrolysis occurred at 0.1 and 0.2 m of urea, respectively. FtsZ lost its functional properties before any significant perturbation in the secondary or tertiary structure was detected by using several fluorescence techniques and far UV-CD indicating preferential local unfolding of the functional region(s). In addition, the unfolded FtsZ regains its ability to polymerize fully upon removal of urea. The data taken together suggest that FtsZ unfolds reversibly through a multistep process, and local responses that inhibit functional properties precede the global transition of FtsZ to the unfolded state. PMID- 12649277 TI - In vivo imaging of the dynamics of glucose uptake in the cytosol of COS-7 cells by fluorescent nanosensors. AB - Glucose homeostasis is a function of glucose supply, transport across the plasma membrane, and metabolism. To monitor glucose dynamics in individual cells, a glucose nanosensor was developed by flanking the Escherichia coli periplasmic glucose/galactose-binding protein with two different green fluorescent protein variants. Upon binding of substrate the FLIPglu-170n sensor showed a concentration-dependent decrease in fluorescence resonance energy transfer between the attached chromophores with a binding affinity for glucose of 170 nm. Fluorescence resonance energy transfer measurements with different sugars indicated a broad selectivity for monosaccharides. An affinity mutant with a Kd of approximately 600 microM was generated, which showed higher substrate specificity, and thus allowed specific monitoring of reversible glucose dynamics in COS-7 cells in the physiological range. At external glucose concentrations between 0.5 and 10 mM, reflecting typical blood levels, free cytosolic glucose concentrations remained at approximately 50% of external levels. The removal of glucose lead to reduced glucose levels in the cell, demonstrating reversibility and visualizing homeostasis. Glucose levels dropped even in the presence of the transport inhibitor cytochalasin B, indicating rapid metabolism. Consistently, the addition of 2-deoxyglucose, which is not recognized by the sensor, affects glucose uptake and metabolism rates. Within the physiological range, glucose utilization, i.e. hexokinase activity, was not limiting. Furthermore, the results show that in COS-7 cells, cytosolic glucose concentrations can vary over at least two orders of magnitude. The glucose nanosensor provides a novel tool with numerous scientific, medical, and environmental applications. PMID- 12649278 TI - Activation of the prolyl hydroxylase oxygen-sensor results in induction of GLUT1, heme oxygenase-1, and nitric-oxide synthase proteins and confers protection from metabolic inhibition to cardiomyocytes. AB - Recently an oxygen-sensing/transducing mechanism has been identified as a family of O2-dependent prolyl hydroxylase domain-containing enzymes (PHD). In normoxia, PHD hydroxylates a specific proline residue that directs the degradation of constitutively synthesized hypoxia-inducible factor-1alpha. During hypoxia, the cessation of hydroxylation of this proline results in less degradation and thus increases hypoxia-inducible factor-1alpha protein levels. In this study we have examined the consequences of activating the PHD oxygen-sensing pathway in cultured neonatal myocytes using ethyl-3,4 dihydroxybenzoate and dimethyloxalylglycine, inhibitors that, similar to hypoxia, inhibit this family of O2-dependent PHD enzymes. Increased glucose uptake and enhanced glycolytic metabolism are classical cellular responses to hypoxia. Ethyl-3,4 dihydroxybenzoate treatment of cardiomyocyte cultures for 24 h increased [3H]deoxy-4-glucose uptake concurrent with an induction of GLUT1 protein. In addition, ethyl-3,4 dihydroxybenzoate, dimethyloxalylglycine, and hypoxia treatments were found to induce protein levels of nitricoxide synthase-2 and heme oxygenase-1, two important cardioregulatory proteins whose expression in response to hypoxic conditions is poorly understood. In conjunction with these changes in gene expression, activation of the PHD oxygen-sensing mechanism was found to preserve myocyte viability in the face of metabolic inhibition with cyanide and 2 deoxyglucose. These results point to a key role for the PHD pathway in the phenotypic changes that are observed in a hypoxic myocyte and may suggest a strategy to pharmacologically induce protection in heart. PMID- 12649279 TI - Mutations in tau gene exon 10 associated with FTDP-17 alter the activity of an exonic splicing enhancer to interact with Tra2 beta. AB - Mutations in the human tau gene leading to aberrant splicing have been identified in FTDP-17, an autosomal dominant hereditary neurodegenerative disorder. Molecular mechanisms by which such mutations cause tau aberrant splicing were not understood. We characterized two mutations in exon 10 of the tau gene, N279K and Del280K. Our results revealed an exonic splicing enhancer element located in exon 10. The activity of this AG-rich splicing enhancer was altered by N279K and Del280K mutations. This exonic enhancer element interacts with human Tra2 beta protein. The interaction between Tra2 beta and the exonic splicing enhancer correlates with the activity of this enhancer element in stimulating splicing. Biochemical studies including in vitro splicing and RNA interference experiments in transfected cells support a role for Tra2 beta protein in regulating alternative splicing of human tau gene. Our results implicate the human tau gene as a target gene for the alternative splicing regulator Tra2 beta, suggesting that Tra2 beta may play a role in aberrant tau exon 10 alternative splicing and in the pathogenesis of tauopathies. PMID- 12649281 TI - An NF-kappaB-dependent transcriptional program is required for collagen remodeling by human smooth muscle cells. AB - Although remodeling of vessels can dramatically alter lumen diameter and clinical sequelae, the molecular mechanisms regulating extracellular matrix turnover and remodeling are still not well understood. To investigate these processes in human smooth muscle, we have compared their culture on monomer and polymerized collagen gels, conditions that mimic some of the features of injured and normal vessels, respectively. We show that culture on polymerized, but not monomer, collagen leads to the activation of the transcription factor NF-kappaB through phosphorylation and degradation of its inhibitor, IkappaBalpha. Coincident with NF-kappaB activation, expression of MMP1, MMP2, and alpha2 integrin increases on polymerized collagen. Specific inhibition of NF-kappaB by retroviral overexpression of wild-type IkappaBalpha or phosphorylation-resistant, IkappaBalpha-stabilized mutant (IkappaBalphaSer32,36/Ala) reverses the increases in MMP1 and alpha2beta1 on polymerized collagen and decreases collagen gel contraction and degradation. However, forced overexpression of alpha2beta1 integrin or MMP1 in smooth muscle cells expressing IkappaBalphaSer32,36/Ala rescues their ability to contract collagen gels. Thus, polymerized collagen induces NF-kappaB-dependent expression of MMP1 and alpha2beta1 integrin, that are required for smooth muscle extracellular matrix remodeling. PMID- 12649280 TI - The C-terminal tail of the M3-muscarinic receptor possesses anti-apoptotic properties. AB - This study investigates the mechanisms by which the muscarinic receptor gene family can protect against apoptosis. Chinese hamster ovary cells transfected with human muscarinic receptor subtypes underwent apoptotic cell death following treatment with the DNA-damaging agent etoposide. Apoptosis was significantly reduced following muscarinic receptor stimulation of cells that were transfected with receptor subtypes that couple to the Gq/11/phospholipase C pathway, namely M1, M3, and M5. No protection was detected in cells transfected with the Gi coupled M2 and M4 receptors. Further analysis of the Gq/11-coupled M3 receptor revealed that truncation of the carboxyl-tail (Delta 565-M3 mutant) removed the ability of the receptor to protect against etoposide-induced cell death. This mutation did not affect the ability of the receptor to signal through the phospholipase C pathway. Furthermore, activation of the Delta 565-M3 receptor resulted in robust activation of the extracellular-regulated kinase (ERK) and c Jun kinase (JNK). The Delta 565-M3 receptor mutant also underwent agonist-driven phosphorylation in a similar manner to the wild-type receptor indicating that the anti-apoptotic effect of the M3 receptor is independent of receptor phosphorylation. Consistent with this was the fact that two M3-muscarinic receptor mutants deficient in agonist-induced receptor phosphorylation were capable of producing a full anti-apoptotic response. We conclude that the anti apoptotic response of the muscarinic receptor family was confined to the Gq/11 coupled members of this family. The direct involvement of Gq/11/phospholipase C signaling and the ERK-1/2 and JNK pathways together with receptor phosphorylation in the anti-apoptotic response were eliminated. Mutation of a poly-basic region within the short C-terminal tail of the M3-muscarinic receptor inhibited the ability of the receptor to induce an anti-apoptotic response. We conclude that the conserved poly-basic region in the C-terminal tail of the M1, M3, and M5 receptors contributes to the ability of these receptors to mediate protection against apoptotic cell death. PMID- 12649282 TI - Vascular endothelial growth factor-dependent down-regulation of Flk-1/KDR involves Cbl-mediated ubiquitination. Consequences on nitric oxide production from endothelial cells. AB - Ligand-stimulated degradation of receptor tyrosine kinase (RTK) is an important regulatory step of signal transduction. The vascular endothelial growth factor (VEGF) receptor Flk-1/KDR is responsible for the VEGF-stimulated nitric oxide (NO) production from endothelial cells. Cellular mechanisms mediating the negative regulation of Flk-1 signaling in endothelial cells have not been investigated. Here we show that Flk-1 is rapidly down-regulated following VEGF stimulation of bovine aortic endothelial cells (BAECs). Consequently, VEGF pretreatment of endothelial cells prevents any further stimulation of Flk-1, resulting in decreased NO production from subsequent VEGF challenges. Ubiquitination of RTKs targets them for degradation; we demonstrate that activation of Flk-1 by VEGF leads to its polyubiquitination in BAECs. Furthermore, VEGF stimulation of BAECs or COS-7 cells transiently transfected with Flk-1 results in the phosphorylation of the ubiquitin ligase Cbl, the enhanced association of Cbl with Flk-1, and the relocalization of Cbl to vesicular structures in BAECs. Overexpression of Cbl in COS-7 cells enhances VEGF induced ubiquitination of Flk-1, whereas a Cbl mutant lacking the ubiquitin ligase RING finger domain, 70Z/3-Cbl, does not. Moreover, expression of Cbl in contrast to 70Z/3-Cbl inhibits the Flk-1-dependent activation of eNOS and, thus, NO release. In BAEC overexpressing Cbl, the degradation of Flk-1 upon VEGF stimulation is accelerated compared with cells transfected with a control vector (green fluorescent protein). Our findings demonstrate that Flk-1 is rapidly down regulated following sustained VEGF stimulation and identify Cbl as a negative regulator of Flk-1 signaling to eNOS. Cbl thus plays a role in the regulation of VEGF signaling by mediating the stimulated ubiquitination and, consequently, degradation of Flk-1 in endothelial cells. PMID- 12649283 TI - Munc18-syntaxin complexes and exocytosis in human platelets. AB - The Sec1-Munc18 (SM) proteins are required for cellular exocytosis, but their mechanistic function remains poorly understood. We examined SM-syntaxin complexes in human platelets, which are terminally differentiated, anuclear cells that secrete the contents of their intracellular granules through syntaxin 2- and syntaxin 4-dependent mechanisms. Munc18a, Munc18b, and Munc18c were detected in human platelets by immunoblotting and/or PCR. The SM proteins and syntaxin 2 were found in the membrane and cytosolic fractions of cells, whereas syntaxin 4 was detected only in the membrane. Platelet membranes contain Munc18c-syntaxin 4 complexes, but minimal if any Munc18c-syntaxin 2 complexes were found. No significant amounts of Munc18a or Munc18b complexes were seen with either syntaxin. Munc18c-syntaxin 4 complexes were dissociated when cells were activated to secrete. Two potential inhibitors of Munc18c-syntaxin 4 complexes were generated to examine whether complex dissociation may lead to exocytosis. Peptides that mimic the projected intermolecular contact sites of Munc18c with syntaxin enhanced Ca2+-triggered dense granule exocytosis in permeabilized cells. Similarly, an anti-Munc18c monoclonal antibody that inhibited the Munc18c syntaxin complex potently amplified Ca2+-induced platelet granule secretion. In summary, Munc18 proteins bind to specific syntaxin isoforms in platelets despite the presence of other potential binding partners. Acute inhibition of the SM syntaxin complex promotes Ca2+-induced exocytosis, suggesting that complex formation per se has a regulatory effect on triggered secretion. PMID- 12649284 TI - Importance of conserved N-domain residues Thr441, Glu442, Lys515, Arg560, and Leu562 of sarcoplasmic reticulum Ca2+-ATPase for MgATP binding and subsequent catalytic steps. Plasticity of the nucleotide-binding site. AB - Nine single mutations were introduced to amino acid residues Thr441, Glu442, Lys515, Arg560, Cys561, and Leu562 located in the nucleotide-binding domain of sarcoplasmic reticulum Ca2+-ATPase, and the functional consequences were studied in a direct nucleotide binding assay, as well as by steady-state and transient kinetic measurements of the overall and partial reactions of the transport cycle. Some partial reaction steps were also examined in mutants with alterations to Phe487, Arg489, and Lys492. The results implicate all these residues, except Cys561, in high affinity nucleotide binding at the substrate site. Mutations Thr441 --> Ala, Glu442 --> Ala, and Leu562 --> Phe were more detrimental to MgATP binding than to ATP binding, thus pointing to a role for these residues in the binding of Mg2+ or to a difference between the interactions with MgATP and ATP. Subsequent catalytic steps were also selectively affected by the mutations, showing the involvement of the nucleotide-binding domain in these reactions. Mutation of Arg560 inhibited phosphoryl transfer but enhanced the E1PCa2 --> E2P conformational transition, whereas mutations Thr441 --> Ala, Glu442 --> Ala, Lys492 --> Leu, and Lys515 --> Ala inhibited the E1PCa2 --> E2P transition. Hydrolysis of the E2P phosphoenzyme intermediate was enhanced in Glu442 --> Ala, Lys492 --> Leu, Lys515 --> Ala, and Arg560 --> Glu. None of the mutations affected the low affinity activation by nucleotide of the phosphoenzyme processing steps, indicating that modulatory nucleotide interacts differently from substrate nucleotide. Mutation Glu442 --> Ala greatly enhanced reaction of Lys515 with fluorescein isothiocyanate, indicating that the two residues form a salt link in the native protein. PMID- 12649285 TI - Nucleotide-dependent triggering of RNA polymerase-DNA interactions by an AAA regulator of transcription. AB - Enhancer-dependent activator proteins, which act upon the bacterial RNA polymerase containing the sigma54 promoter specificity factor, belong to the AAA superfamily of ATPases. Activator-sigma54 contact is required for the sigma54 RNAP to isomerize and engage the DNA template for transcription. How ATP hydrolysis is used to trigger changes in sigma54-RNA polymerase and promoter DNA that lead to DNA opening is poorly understood. Here, band shift and footprinting assays were used to investigate the DNA binding activities of sigma54 and sigma54 RNA polymerase in the presence of the activator protein PspF bound to poorly hydrolysable analogues of ATP and the ATP hydrolysis transition-state analogue ADP.AlFx. Results show that different nucleotide-bound forms of PspF can change the interactions between sigma54, sigma54-RNA polymerase, and a DNA fork junction structure present within closed promoter complexes. This provides evidence that in the activation transduction pathway, several functional states of the activator, prior to ATP hydrolysis, can serve to alter the fork junction binding activity of sigma54 and sigma54-RNA polymerase that precede full DNA opening. A sequential set of nucleotide-dependent transitions in sigma54-RNA polymerase promoter complexes needed for productive open complex formation may therefore depend upon different nucleotide-bound forms of the activator. PMID- 12649286 TI - Architecture of the replication complex and DNA loops at the fork generated by the bacteriophage t4 proteins. AB - Rolling circle replication has previously been reconstituted in vitro using M13 duplex circles containing preformed forks and the 10 purified T4 bacteriophage replication proteins. Leading and lagging strand synthesis in these reactions is coupled and the size of the Okazaki fragments produced is typical of those generated in T4 infections. In this study the structure of the DNAs and DNA protein complexes engaged in these in vitro reactions has been examined by electron microscopy. Following deproteinization, circular duplex templates with linear tails as great as 100 kb are observed. The tails are fully duplex except for one to three single-stranded DNA segments close to the fork. This pattern reflects Okazaki fragments stopped at different stages in their synthesis. Examination of the DNA-protein complexes in these reactions reveals M13 duplex circles in which 64% contain a single large protein mass (replication complex) and a linear duplex tail. In 56% of the replicating molecules with a tail there is at least one fully duplex loop at the replication complex resulting from the portion of the lagging strand engaged in Okazaki fragment synthesis folding back to the replisome. The single-stranded DNA segments at the fork bound by gene 32 and 59 proteins are not extended but rather appear organized into highly compact structures ("bobbins"). These bobbins constitute a major portion of the mass of the full replication complex. PMID- 12649287 TI - Potent inhibition of ribonuclease A by oligo(vinylsulfonic acid). AB - Ribonuclease A (RNase A) can make multiple contacts with an RNA substrate. In particular, the enzymatic active site and adjacent subsites bind sequential phosphoryl groups in the RNA backbone through Coulombic interactions. Here, oligomers of vinylsulfonic acid (OVS) are shown to be potent inhibitors of RNase A that exploit these interactions. Inhibition is competitive with substrate and has Ki = 11 pm in assays at low salt concentration. The effect of salt concentration on inhibition indicates that nearly eight favorable Coulombic interactions occur in the RNase A.OVS complex. The phosphonic acid and sulfuric acid analogs of OVS are also potent inhibitors although slightly less effective. OVS is also shown to be a contaminant of MES and other buffers that contain sulfonylethyl groups. Oligomers greater than nine units in length can be isolated from commercial MES buffer. Inhibition by contaminating OVS is responsible for the apparent decrease in catalytic activity that has been observed in assays of RNase A at low salt concentration. Thus, OVS is both a useful inhibitor of RNase A and a potential bane to chemists and biochemists who use ethanesulfonic acid buffers. PMID- 12649288 TI - Inactivation of menin, the product of the multiple endocrine neoplasia type 1 gene, inhibits the commitment of multipotential mesenchymal stem cells into the osteoblast lineage. AB - The physiological roles of menin, the product of the multiple endocrine neoplasia type 1 gene, are not known. Homozygous menin knockout mice exhibit cranial and facial hypoplasia. We, therefore, investigated the role of menin in the regulation of osteoblastic differentiation. Menin antisense oligonucleotides (AS oligo) reduced endogenous menin expression in the C3H10T1/2 (10T1/2) mouse mesenchymal stem cells and antagonized alkaline phosphatase (ALP) activity and the expression of type I collagen, Runx2/cbfa1 (Runx2), and osteocalcin (OCN) induced by bone morphogenetic protein 2 (BMP-2). AS-oligo did not affect adipogenic markers (Oil red staining and PPARgamma expression) and chondrogenic markers (Alcian blue staining and type IX collagen) induced by BMP-2 in 10T1/2 cells. Menin co-immunoprecipitated with Smad1 and Smad5, and inactivation of menin antagonized BMP-2-induced transcriptional activity of Smad1/5. In osteoblastic MC3T3-E1 cells, AS-oligo affected neither BMP-2-stimulated ALP activity nor the expression of Runx2 and OCN. Stable inactivation of menin in MC3T3-E1 cells increased ALP activity, mineralization, and the expression of type I collagen and OCN. In 21-day cultures of MC3T3-E1 cells and BMP-2-treated 10T1/2 cells, endogenous menin expression increased up to day 14 and declined thereafter. These data indicate that menin inactivation specifically inhibits the commitment of pluripotent mesenchymal stem cells to the osteoblast lineage, mediated by menin and Smad1/5 interactions. Menin is important for both early differentiation of osteoblasts and inhibition of their later differentiation, and it might be crucial for intramembranous ossification. PMID- 12649289 TI - Reversible glutathionylation of complex I increases mitochondrial superoxide formation. AB - Increased production of reactive oxygen species (ROS) by mitochondria is involved in oxidative damage to the organelle and in committing cells to apoptosis or senescence, but the mechanisms of this increase are unknown. Here we show that ROS production by mitochondrial complex I increases in response to oxidation of the mitochondrial glutathione pool. This correlates with thiols on the 51- and 75 kDa subunits of complex I forming mixed disulfides with glutathione. Glutathionylation of complex I increases superoxide production by the complex, and when the mixed disulfides are reduced, superoxide production returns to basal levels. Within intact mitochondria oxidation of the glutathione pool to glutathione disulfide also leads to glutathionylation of complex I, which correlates with increased superoxide formation. In this case, most of this superoxide is converted to hydrogen peroxide, which can then diffuse into the cytoplasm. This mechanism of reversible mitochondrial ROS production suggests how mitochondria might regulate redox signaling and shows how oxidation of the mitochondrial glutathione pool could contribute to the pathological changes that occur to mitochondria during oxidative stress. PMID- 12649290 TI - The a-subunit of the V-type H+-ATPase interacts with phosphofructokinase-1 in humans. AB - V-type or H+-ATPases are a family of ATP-dependent proton pumps that move protons across the plasma membrane at specialized sites such as kidney epithelial cells and osteoclasts as well as acidifying intracellular compartments. The 100-kDa polytopic a-subunit of this group of ATPases is suggested to play an important role in coupling the two functions of the pump, ATP hydrolysis and proton transport. In man, different a-subunit isoforms are encoded by four genes. ATP6V0A4 encodes a4, which is expressed apically in alpha-intercalated cells in both human and mouse kidney. We sought binding partners for the C terminus of a4 in order to address its potential role in the H+-ATPase complex. Random peptide phage display analysis revealed a consensus motif (WLELRP) with almost complete homology to part of the enzyme phosphofructokinase 1 (PFK-1). Activity of this enzyme is the rate-limiting step in glycolysis. Specificity of a4 binding to this peptide was confirmed by enzyme-linked immunosorbent assay. Protein-protein interaction was further demonstrated by co-immunoprecipitation of a4 with PFK-1 from solubilized human kidney membrane proteins. An in vitro bead-bound PFK-1 pull-down assay showed that this interaction was also true for the ubiquitously expressed a1 subunit. Finally, PFK-1 co-immunolocalized with a4 in alpha intercalated cells in the collecting ducts of human kidney. These findings indicate a direct link between V-type H+-ATPases and glycolysis via the C terminal region of the a-subunit of the pump and suggest a novel regulatory mechanism between H+-ATPase function and energy supply. This interaction between the a-subunit and PFK-1 also provides new evidence that the C terminus of this subunit lies cytoplasmically in vivo. PMID- 12649291 TI - Regulatory role of arginine 204 in the catalytic activity of rat alloantigens ART2a and ART2b. AB - ART2a (RT6.1) and ART2b (RT6.2) are NAD glycohydrolases (NADases) that are linked to T lymphocytes by glycosylphosphatidylinositol anchors. Although both mature proteins possess three conserved regions (I, II, III) that form the NAD-binding site and differ by only ten amino acids, only ART2b is auto-ADP-ribosylated and only ART2a is glycosylated. To investigate the structural basis for these differences, wild-type and mutant ART2a and ART2b were expressed in rat mammary adenocarcinoma (NMU) cells and released with phosphatidylinositol-specific phospholipase C. All mutants were immunoreactive NADases. Arginine 204 (Arg204), NH2-terminal to essential glutamate 209 in Region III, is found in ART2b, but not ART2a. Replacement of Arg204 in ART2b with lysine, tyrosine, or glutamate abolished auto-ADP-ribosylation. Unlike wild-type ART2a, ART2a(Y204R) was auto ADP-ribosylated. The tryptophan mutant ART2b(R204W) was auto-ADP-ribosylated and exhibited enhanced NADase activity. Incubation with NAD and auto-ADP-ribosylation decreased the NADase activities of wild-type ART2b and ART2b (R204W), whereas activity of ART2b(R204K), which is not auto-modified, was unchanged by NAD. Facilitation of auto-ADP-ribosylation by tryptophan 204 suggests that the hydrophobic amino acid mimics an ADP-ribosylated arginine. Thus, Arg204 in ART2b serves as a regulatory switch whose presence is required for additional auto-ADP ribosylation and regulation of catalytic activity. PMID- 12649292 TI - A novel mode of polymerization of alpha1-proteinase inhibitor. AB - Patients homozygous for the Z mutant form of alpha1-proteinase inhibitor (alpha1 PI) have an increased risk for the development of liver disease because of the accumulation in hepatocytes of inclusion bodies containing linear polymers of mutant alpha1-PI. The most widely accepted model of polymerization proposes that a linear, head-to-tail polymer forms by sequential insertion of the reactive center loop (RCL) of one alpha1-PI monomer between the central strands of the A beta-sheet of an adjacent monomer. This model derives primarily from two observations: peptides that are homologous with the RCL insert into the A beta sheet of alpha1-PI monomer and this insertion prevents alpha1-PI polymerization. Normal alpha1-PI monomer does not spontaneously polymerize; however, here we show that the disulfide-linked dimer of normal alpha1-PI spontaneously forms linear polymers in buffer. The monomers within this dimer are joined head-to-head. Thus, the arrangement of monomers in these polymers must be different from that predicted by the loop-A sheet model. Therefore, we propose a new model for alpha1 PI polymer. In addition, polymerization of disulfide-linked dimer is not inhibited by the presence of the peptide even though dimer appears to interact with the peptide. Thus, RCL insertion into A beta-sheets may not occur during polymerization of this dimer. PMID- 12649293 TI - Reducing the agonist activity of antiandrogens by a dominant-negative androgen receptor coregulator ARA70 in prostate cancer cells. AB - Although the progression of prostate cancer initially is dependent on androgens, tumor progression to an androgen-independent growth eventually occurs in most of patients treated with androgen ablation and/or antiandrogen therapy. After the initial response, antiandrogens lose their efficacy and eventually act as agonists to promote androgen receptor (AR)-mediated growth of prostate cancer cells. An aberrant regulation of AR activity, presumably by AR coregulators, may contribute to this acquired agonist activity of antiandrogens. Using an in vitro mutagenesis and a double-negative selection in yeast two-hybrid screening, we have identified a dominant-negative AR coregulator ARA70 (dARA70N), which can inhibit AR transcriptional activity by inactivating the normal function of ARA70 in the LNCaP cells. Whereas ARA70 in oligomeric form interacts with AR and enhances its transcriptional activity, dARA70N lacks AR interaction and might retain the ability to form a non-functional heteromer with ARA70 and interrupt AR transcriptional activity without a change in AR protein itself. The addition of dARA70N reduces the agonist activity and rescues the normal function of antiandrogens in prostate cancer cells. RNA-interference-mediated silencing of ARA70 gene further confirms these observations. Taken together, these findings indicate that ARA70 may contribute to the acquired agonist activity of antiandrogens and plays an important role in making prostate cancer cells resistant to androgen ablation and/or antiandrogen therapy. ARA70 may, thus, be a critical target for developing therapeutic agents against AR-mediated progression of prostate cancer. PMID- 12649294 TI - Rat inositol 1,4,5-trisphosphate 3-kinase C is enzymatically specialized for basal cellular inositol trisphosphate phosphorylation and shuttles actively between nucleus and cytoplasm. AB - The calcium-liberating second messenger inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) is converted to inositol 1,3,4,5-tetrakisphosphate (Ins(1,3,4,5)P4) by Ins(1,4,5)P3 3-kinases (IP3Ks) that add a fourth phosphate group to the 3-position of the inositol ring. Two isoforms of IP3Ks (named A and B) from different vertebrate species have been well studied. Recently the cloning and examination of a human full-length cDNA encoding a novel isoform, termed human IP3K-C (HsIP3K-C), has been reported. In the present study we report the cloning of a full-length cDNA encoding a rat homologue of HsIP3K-C with a unique mRNA expression pattern, which differs remarkably from the tissue distribution of HsIP3K-C. Of the rat tissues examined, rat IP3K-C (RnIP3K-C) is mainly present in heart, brain, and testis and shows the strongest expression in an epidermal tissue, namely tongue epithelium. RnIP3K-C has a calculated molecular mass of approximately 74.5 kDa and shows an overall identity of approximately 75% with HsIP3K-C. A bacterially expressed, enzymatically active and Ca2+-calmodulin regulated fragment of this isoform displays remarkable enzymatic properties like a very low Km for Ins(1,4,5)P3 ( approximately 0.2 microm), substrate inhibition by high concentrations of Ins(1,4,5)P3, allosteric product activation by Ins(1,3,4,5)P4 in absence of Ca2+-calmodulin (Ka(app) 0.52 microm), and the ability to efficiently phosphorylate a second InsP3 substrate, inositol 2,4,5 trisphosphate, to inositol 2,4,5,6-tetrakisphosphate in the presence of Ins(1,3,4,5)P4. Furthermore, the RnIP3K-C fused with a fluorescent protein tag is actively transported into and out of the nucleus when transiently expressed in mammalian cells. A leucine-rich nuclear export signal and an uncharacterized nuclear import activity are localized in the N-terminal domain of the protein and determine its nucleocytoplasmic shuttling. These findings point to a particular role of RnIP3K-C in nuclear inositol trisphosphate phosphorylation and cellular growth. PMID- 12649295 TI - Total conversion of bifunctional catalase-peroxidase (KatG) to monofunctional peroxidase by exchange of a conserved distal side tyrosine. AB - Catalase-peroxidases (KatGs) are unique peroxidases exhibiting a high catalase activity and a peroxidase activity with a wide range of artificial electron donors. Exchange of tyrosine 249 in Synechocystis KatG, a distal side residue found in all as yet sequenced KatGs, had dramatic consequences on the bifunctional activity and the spectral features of the redox intermediate compound II. The Y249F variant lost catalase activity but retained a peroxidase activity (substrates o-dianisidine, pyrogallol, guaiacol, tyrosine, and ascorbate) similar to the wild-type protein. In contrast to wild-type KatG and similar to monofunctional peroxidases, the formation of the redox intermediate compound I could be followed spectroscopically even by addition of equimolar hydrogen peroxide to ferric Y249F. The corresponding bimolecular rate constant was determined to be (1.1 +/- 0.1) x 107 m-1 s-1 (pH 7 and 15 degrees C), which is typical for most peroxidases. Additionally, for the first time a clear transition of compound I to an oxoferryl-like compound II with peaks at 418, 530, and 558 nm was monitored when one-electron donors were added to compound I. Rate constants of reaction of compound I and compound II with tyrosine ((5.0 +/- 0.3) x 104 m-1 s-1 and (1.7 +/- 0.4) x 102 m-1 s-1) and ascorbate ((1.3 +/- 0.2) x 104 m-1 s-1 and (8.8 +/- 0.1) x 101 m-1 s-1 at pH 7 and 15 degrees C) were determined by using the sequential stopped-flow technique. The relevance of these findings is discussed with respect to the bifunctional activity of KatGs and the recently published first crystal structure. PMID- 12649296 TI - Galantamine is an allosterically potentiating ligand of neuronal nicotinic but not of muscarinic acetylcholine receptors. AB - Galantamine (Reminyl), an approved treatment for Alzheimer's disease (AD), is a potent allosteric potentiating ligand (APL) of human alpha 3 beta 4, alpha 4 beta 2, and alpha 6 beta 4 nicotinic receptors (nAChRs), and of the chicken/mouse chimeric alpha 7/5-hydroxytryptamine3 receptor, as was shown by whole-cell patch clamp studies of human embryonic kidney-293 cells stably expressing a single nAChR subtype. Galantamine potentiates agonist responses of the four nAChR subtypes studied in the same window of concentrations (i.e., 0.1-1 microM), which correlates with the cerebrospinal fluid concentration of the drug at the recommended daily dosage of 16 to 24 mg. At concentrations >10 microM, galantamine acts as an nAChR inhibitor. The other presently approved AD drugs, donepezil and rivastigmine, are devoid of the nicotinic APL action; at micromolar concentrations they also block nAChR activity. Using five CHO-SRE-Luci cell lines, each of them expressing a different human muscarinic receptor, and a reporter gene assay, we show that galantamine does not alter the activity of M1 M5 receptors, thereby confirming that galantamine modulates selectively the activity of nAChRs. These studies support our previous proposal that the therapeutic action of galantamine is mainly produced by its sensitizing action on nAChRs rather than by general cholinergic enhancement due to cholinesterase inhibition. Galantamine's APL action directly addresses the nicotinic deficit in AD. PMID- 12649297 TI - Use of irreversible antagonists to determine the relative efficacy of mu-opioids in a pigeon drug discrimination procedure: comparison of beta-funaltrexamine and clocinnamox. AB - The use of irreversible antagonists to assess opioid efficacy has proven fruitful for classifying opioids on the basis of high or low efficacy, but few studies have provided quantitative estimates of efficacy. The purpose of this study was to use beta-funaltrexamine (beta-FNA) and clocinnamox (C-CAM) in a drug discrimination procedure to examine the efficacy of fentanyl, morphine, l methadone, sufentanil, and etorphine. In pigeons trained to discriminate 0.12 mg/kg fentanyl from water, dose-effect curves were determined for each opioid alone and after pretreatment with beta-FNA and C-CAM. Using quantitative analyses according to an extended model of Black and Leff (1983), apparent efficacy (tau) and affinity (KA) of each opioid was determined, as well as the degree of receptor inactivation (q) produced by each dose of each antagonist. beta-FNA and C-CAM produced dose- and time-dependent, rightward shifts in the dose-effect curves of each opioid, and analyses based on dose-ratios and tau values suggest a rank order of efficacy of etorphine > sufentanil = l-methadone > fentanyl = morphine. Marked differences in the profiles of antagonism produced by beta-FNA and C-CAM were also apparent, as C-CAM, but not beta-FNA, produced insurmountable antagonism. The q values for each antagonist were consistent with these data in indicating that C-CAM and beta-FNA can inactivate nearly 100 and 75% of the receptor population, respectively. In tests conducted in pigeons chronically treated with morphine, doses of beta-FNA that produced parallel, rightward shifts in untreated pigeons flattened the morphine dose-effect curve in morphine-treated pigeons. These results indicate that beta-FNA and C-CAM can differentiate opioids with high relative efficacy and yield comparable estimates of efficacy for various opioids. There are, however, limitations in the proportion of the receptor population that can by eliminated by beta-FNA. PMID- 12649298 TI - A novel free radical scavenger, edarabone, protects against cisplatin-induced acute renal damage in vitro and in vivo. AB - Accumulating evidence suggests that enhanced peroxidative damage caused by reactive oxygen species (ROS) may contribute to the pathogenesis of cisplatin induced acute renal failure. Nevertheless, little is known about the involvement of oxygen radicals in cisplatin nephropathy. In this study, we investigated the effects of a novel free radical scavenger, 3-methyl-1-phenyl-pyrazolin-5-one (MCI 186; edarabone), on murine proximal tubular cell (PTC) damage induced by exposure to cisplatin in vitro and on renal function in an in vivo model of cisplatin induced acute renal failure. Edarabone inhibited cisplatin-induced (40 microM, 24 h) cytotoxicity in a concentration-dependent manner (10-5 to 10-3 M). Edarabone also attenuated cisplatin-induced mitochondrial transmembrane potential loss and ROS production of PTCs. In the in vivo study, male Wistar rats were cotreated with cisplatin (5 mg/kg, i.p.) and edarabone (1 or 5 mg/kg, i.v.). Effects of edarabone on the kidney were examined 5 days after treatment. Cisplatin resulted in renal dysfunction, renal tubular damage, mitochondrial damage (assayed by histochemical staining for respiratory chain complex IV), renal protein oxidation (examined by Western blot analysis using a specific antibody for carbonyl group containing proteins), and tubular apoptosis (determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling staining). The above changes were attenuated by edarabone treatment. Thus, edarabone exhibited cytoprotective effects in PTCs and renoprotective effects against cisplatin. Our findings suggest that ROS, in particular hydroxyl radicals, are involved in cisplatin nephropathy and that edarabone may be potentially useful in protecting the kidneys and prevention of acute renal failure. PMID- 12649299 TI - Nicotinic receptors differentially regulate N-methyl-D-aspartate damage in acute hippocampal slices. AB - Although in neuronal cultures nicotine was reported to prevent early and delayed excitotoxic death, no studies with nicotinic drugs have been done with acute hippocampal slices. We investigated the effect of nicotine and methyllycaconitine (MLA) on the toxicity of N-methyl-d-aspartate (NMDA) in the CA1 area of hippocampal slices. The excitotoxic effect of NMDA was assessed as decreased recovery of the capability to produce synaptically evoked population spikes (PSs). Application of nicotine or MLA before NMDA application increased the recovery of PSs. This electrophysiological recovery was used as a measure of the early neuroprotective events. The neuroprotection conferred by both nicotine and MLA was inhibited by dihydro-beta-erythroidine, showing mediation of neuroprotection by alpha 4 beta 2 neuronal nicotinic receptors (nAChRs). Because nicotine activates alpha 4 beta 2 and other nAChR subtypes, whereas 10 nM MLA inhibits the alpha 7 subtype, we propose the involvement of a neuronal circuitry dependent mechanism for nicotinic neuroprotection. The effect of nicotine downstream from the receptors was investigated using inhibitors of cell signaling. The results suggest that the effect of nicotine is mediated by tyrosine receptor kinases, 1,2-phosphatidylinositol-3 kinase, and the mitogen activated extracellular signal-regulated kinases. Although nicotine neuroprotection is Ca2+-dependent, neither L-type Ca2+ channels nor calmodulin dependent protein kinase is involved in the effect of nicotine. In summary, these results suggest that in acute slices nicotinic protection is initiated either by direct activation of alpha 4 beta 2 or indirectly by inhibition of alpha 7 followed by signal transduction involving tyrosine kinases, phospholipid dependent kinases, and mitogen-activated kinases. PMID- 12649300 TI - Dissimilar pharmacological responses by a new series of imidazoline derivatives at precoupled and ligand-activated alpha 2A-adrenoceptor states: evidence for effector pathway-dependent differential antagonism. AB - Whereas agonist-directed differential signaling at a single receptor subtype has become an accepted pharmacological concept, distinct behaviors by ligands that are assumed to be antagonists is less documented. The intrinsic activity and capacity of antagonism for a new series of imidazoline-derived adrenergic ligands analogous to dexefaroxan were investigated by measuring two distinct signaling pathways at the recombinant human alpha 2A-adrenoceptor (alpha 2A AR): 1) pertussis toxin-resistant guanosine 5'-O-(3-[35S]thio)triphosphate ([35S]GTP gamma S) binding responses mediated by either a recombinant G alpha oCys351Ile or G alpha i2Cys352Ile protein in CHO-K1 cells, and 2) inhibition of cAMP formation in a stably transfected C6-glial cell line. Ligands could be differentiated as inverse agonists [i.e., 2-(4-methoxy-2-ethyl-2,3-dihydrobenzofuran-2-yl)-4,5 dihydro-1H-imidazole; RX 851062], neutral antagonists [i.e., 2-(4-hydroxy-2-ethyl 2,3-dihydrobenzofuran-2-yl)-4,5-dihydro-1H-imidazole; RX 851057], partial [i.e., 2-(4-chloro-2,3-dihydrobenzofuran-2-yl)-4,5-dihydro-1H-imidazole; RX 821008], and high-efficacy [i.e., 2-(6,7-dichloro-2,3-dihydrobenzofuran-2-yl)-4,5-dihydro-1H imidazole; RX 821010] agonists at a precoupled alpha 2A AR state in the copresence of a G alpha oCys351Ile protein but not G alpha i2Cys352Ile protein by monitoring [35S]GTP gamma S binding responses. Neither positive nor negative efficacy was observed for these compounds by monitoring the adenylate cyclase pathway at a presumably low-affinity alpha 2A AR state. The capacity of the dexefaroxan analogs to antagonize the (-)-epinephrine-mediated [35S]GTP gamma S binding response at a G alpha oCys351Ile protein was inversely correlated with their magnitude of intrinsic activity and unrelated to their ligand binding affinity for the alpha 2A AR. On the other hand, their capacity to antagonize either (-)-epinephrine or 5-bromo-6-(2-imidazolin-2-ylamino)quinoxaline tartrate (UK 14304)-mediated inhibition of forskolin-stimulated cAMP formation was not related with the rank order of antagonist capacity for the (-)-epinephrine mediated [35S]GTP gamma S binding response. In conclusion, these data demonstrate that certain alpha2 AR ligands that are assumed to be antagonists, may yield dissimilar pharmacological responses, dependent on the investigated agonist stimulated effector pathway. PMID- 12649301 TI - Enhancement of cytokine production and AP-1 transcriptional activity in T cells by thalidomide-related immunomodulatory drugs. AB - CC-4047 (Actimid) and CC-5013 (Revimid) belong to a class of thalidomide analogs collectively known as the immunomodulatory drugs (IMiDs), which are currently being assessed in the treatment of patients with multiple myeloma and other cancers. IMiDs potently enhance T cell and natural killer cell responses and inhibit tumor necrosis factor-alpha, interleukin (IL)-1 beta, and IL-12 production from LPS-stimulated peripheral blood mononuclear cells. However, the molecular mechanism of action for these compounds is unknown. Herein, we report on the ability of the IMiDs to up-regulate production of IL-2 from activated human CD4+ and CD8+ peripheral blood T cells, production of IL-2 and IFN-gamma from T helper (Th)1-type cells, and production of IL-5 and IL-10 from Th2-type cells. Elevation of IL-2 production from Jurkat T cells was observed as early as 6 h poststimulation and correlated with an increase in IL-2 promoter activity that was dependent upon the proximal but not the distal AP-1 binding site. The IMiDs enhanced AP-1-driven transcriptional activity 2- to 4-fold after 6 h of T cell stimulation, and their relative potencies for AP-1 activation correlated with their potencies for increased IL-2 production in Jurkat T cells and in CD4+ or CD8+ human peripheral blood T cells. The most potent of these IMiDs, CC-4047, had no effect on nuclear factor of activated T cells transcriptional activity, calcium signaling, or phosphorylation of extracellular signal-regulated kinase 1/2, c-Jun NH2-terminal kinase 1/2, p38 mitogen-activated protein kinase, or c Jun/Jun D in Jurkat T cells. These data suggest that IMiDs increase T cell cytokine production by potentiating AP-1 transcriptional activity. PMID- 12649302 TI - Differential cardiovascular regulatory activities of the alpha 1B- and alpha 1D adrenoceptor subtypes. AB - The regulation of cardiac and vascular function by the alpha 1B- and alpha 1D adrenoceptors (ARs) has been assessed in two lines of transgenic mice, one over expressing a constitutively active alpha 1B-AR mutation (alpha 1B-ARC128F) and the other an alpha 1D-AR knockout line. The advantage of using mice expressing a constitutively active alpha 1B-AR is that the receptor is tonically active, thus avoiding the use of nonselective agonists that can activate all subtypes. In hearts from animals expressing alpha 1B-ARC128F, the activities of the mitogen activated protein kinases, extracellular signal-regulated kinase, and c-Jun N terminal kinase were significantly elevated compared with nontransgenic control animals. Mice over-expressing the alpha 1B-ARC128F had echocardiographic evidence of contractile dysfunction and increases in chamber dimensions. In isolated perfused hearts or left ventricular slices from alpha 1B-ARC128F-expressing animals, the ability of isoproterenol to increase contractile force or increase cAMP levels was significantly decreased. In contrast to the prominent effects on the heart, constitutive activation of the alpha 1B-AR had little effect on the ability of phenylephrine to induce vascular smooth muscle contraction in the isolated aorta. The ability of phenylephrine to stimulate coronary vasoconstriction was diminished in alpha 1D-AR knockout mice. In alpha 1D-AR knockout animals, no negative effects on cardiac contractile function were noted. These results show that the alpha1-ARs regulate distinctly different physiologic processes. The alpha 1B-AR appears to be involved in the regulation of cardiac growth and contractile function, whereas the alpha 1D-AR is coupled to smooth muscle contraction and the regulation of systemic arterial blood pressure. PMID- 12649303 TI - Peroxisome proliferator-activated receptor ligands affect growth-related gene expression in human leukemic cells. AB - Peroxisome proliferator-activated receptors (PPARs) are ligand-activated nuclear receptors. Three subtypes of PPARs (alpha, beta, and gamma) have been identified in different tissues. PPAR alpha and PPAR gamma ligands inhibit cell proliferation and induce differentiation in several human cell models. We demonstrated that both PPAR alpha (clofibrate and ciprofibrate) and PPAR gamma ligands (troglitazone and 15 deoxy-prostaglandin J2, 15d-PGJ2) inhibited growth, induced the onset of monocytic-like differentiation, and increased the proportion of G0/G1 cells in the HL-60 leukemic cell line. Moreover, 3 days after the treatment with 2.5 microM 15d-PGJ2, an increase in sub-G0/G1 population occurred, compatible with an induction of programmed cell death. To clarify the mechanisms involved in HL-60 growth inhibition due to the effects of PPAR ligands, we investigated their action on the expression of some genes involved in the control of cell proliferation, differentiation, and cell cycle progression such as c-myc, c-myb, and cyclin D1 and D2. Clofibrate (50 microM), ciprofibrate (50 microM), and 15d-PGJ2 (2.5 microM) inhibited c-myb and cyclin D2 expression, whereas they did not affect c-myc and cyclin D1 expression. Only troglitazone (5 microM) decreased c-myc mRNA and protein levels, besides decreasing c-myb and cyclin D2. The down-regulations of c-myb and cyclin D2 expression represent the first evidence of the inhibitory effect exerted by PPAR ligands on these genes. Moreover, the inhibition of c-myc expression by troglitazone may depend on a PPAR independent mechanism. PMID- 12649304 TI - Nucleotide-induced restoration of conjunctival chloride and fluid secretion in adenovirus type 5-infected pigmented rabbit eyes. AB - We evaluated the role of extracellular UTP and other nucleotides in the regulation of chloride (JCl) and fluid secretion (JCl) across the pigmented rabbit conjunctiva. Jv was determined in freshly excised conjunctival tissues mounted between two buffer reservoirs maintained in an enclosed environment at 37 degrees C. Short circuit current (Isc) and 36Cl flux were measured using modified Ussing-type chambers. Fluid flux measurements were made with a pair of capacitance probes. After observing the baseline for 15 to 30 min, fluid flux was measured in the presence of mucosally applied nucleotides (10 microM) for a period of 30 min. Mucosal application of 10 microM each of UTP, UDP, ATP, ADP, AMP, adenosine, and ATP-gamma-S transiently stimulated fluid secretion across the conjunctiva to a significant extent for 10 to 15 min. Other nucleotides did not show any significant effect. The stimulation of fluid secretion correlated well with the stimulation in Isc (r2 = 0.85). UTP (0.1-1000 microM) led to a maximal increase in fluid secretion by 11.72 +/- 0.48 microl/(h x cm2) with an EC50 value of 10.39 +/- 1.08 microM. ATP (0.1-1000 microM) caused a maximal increase in fluid secretion by 11.89 +/- 0.88 microl/(h x cm2) with an EC50 value of 17.23 +/ 2.63 microM. Adenovirus type 5 (Ad5) infection significantly decreased both net 36Cl secretion across the conjunctiva by approximately 56% and the rate of fluid secretion by approximately 56%. UTP (10 microM), but not 1 mM 8-bromo-cAMP, was able to elicit a normal stimulatory response in the Ad5-infected tissues. In conclusion, mucosal application of purinergic nucleotides may be therapeutically important in restoring ion and fluid secretion in the diseased conjunctiva. PMID- 12649305 TI - Pharmacological characterization of the novel histamine H3-receptor antagonist N (3,5-dichlorophenyl)-N'-[[4-(1H-imidazol-4-ylmethyl)phenyl]-methyl]-urea (SCH 79687). AB - We present the pharmacological and pharmacokinetic profiles of a novel histamine H3 receptor antagonist, N-(3,5-dichlorophenyl)-N'-[[4-(1H-imidazol-4 ylmethyl)phenyl]-methyl]-urea (SCH 79687). The H3-receptor binding Ki values for SCH 79687 were 1.9 and 13 nM in the rat and guinea pig (GP), respectively. The Ki values for SCH 79687 at histamine H1 and H2 receptors were greater than 1 microM. SCH 79687 showed a 41- and 82-fold binding selectivity for the H3 receptor over alpha 2A-adrenoceptors and imidazoline I2, and >500-fold H3 selectivity compared with over 60 additional receptors. The pA2 value for SCH 79687 in the GP ileum electrical field-stimulated (EFS) contraction was 9.6 +/- 0.3. Similar H3 antagonist activity was observed in the EFS cryopreserved and fresh tissue isolated human saphenous vein (HSV) assays (pKb = 9.4 +/- 0.3 and 10.1 +/- 0.4). SCH 79687 (30 nM) did not block clonidine-induced inhibition of EFS-induced contractions in HSV. SCH 79687 (ED50 = 0.3 mg/kg i.v.) attenuated (R)-alpha methylhistamine inhibition of sympathetic hypertensive responses in the GP. At the time of activity evaluation, the GP plasma SCH 79687 concentration was 25 ng/ml at the dose of 0.3 mg/kg i.v. In feline nasal studies, combined administration of SCH 79687 (3 mg/kg i.v.) and the H1-antagonist loratadine (3 mg/kg i.v.), at individual doses that do not produce decongestion, inhibited the compound 48/80-induced congestion by 47%. The alpha-adrenergic agonist phenylpropanolamine (PPA; 1 mg/kg i.v.) also attenuated compound 48/80 nasal responses by 42%. Unlike the H3/H1 combination that did not affect blood pressure (BP), PPA (1 mg/kg i.v.) significantly increased BP compared with control animals by a maximum of 31 mm Hg. Orally, SCH 79687 (10 mg/kg) plus loratadine (10 mg/kg) also produced decongestion without effects on BP. In pharmacokinetic studies, oral dosing with SCH 79687 in the rat (10 mg/kg) and monkey (3 mg/kg) achieved plasma Cmax and area under the curve values greater than 1.5 and 12.1 microg. h/ml, respectively. SCH 79687 is an orally active H3 antagonist with a good pharmacokinetic profile that, in combination with an H1 antagonist, demonstrates decongestant efficacy comparable with oral sympathomimetic decongestants but without hypertensive liabilities. PMID- 12649306 TI - Sources and significance of plasma levels of catechols and their metabolites in humans. AB - Human plasma contains several catechols, including the catecholamines norepinephrine, epinephrine, and dopamine, their precursor, L-3,4 dihydroxyphenylalanine (L-DOPA), and their deaminated metabolites, dihydroxyphenylglycol, the main neuronal metabolite of norepinephrine, and dihydroxyphenylacetic acid, a deaminated metabolite of dopamine. Products of metabolism of catechols include 3-methoxytyrosine (from L-DOPA), homovanillic acid and dopamine sulfate (from dopamine), normetanephrine, vanillylmandelic acid, and methoxyhydroxyphenylglycol (from norepinephrine), and metanephrine (from epinephrine). Plasma levels of catechols and their metabolites have related but distinct sources and therefore reflect different functions of catecholamine systems. This article provides an update about plasma levels of catechols and their metabolites and the relevance of those levels to some issues in human health and disease. PMID- 12649307 TI - (+)-Fenfluramine and its major metabolite, (+)-norfenfluramine, are potent substrates for norepinephrine transporters. AB - (+/-)-Fenfluramine is an amphetamine analog that was once widely prescribed as an appetite suppressant. Although (+/-)-fenfluramine is no longer clinically available, the mechanisms underlying its anorectic properties are still of interest. Upon peripheral administration, stereoisomers of (+/-)-fenfluramine are N-deethylated to form the metabolites, (+)- and (-)-norfenfluramine. It is well accepted that isomers of (+/-)-fenfluramine and (+/-)-norfenfluramine interact with 5-hydroxytryptamine (serotonin, 5-HT) transporters to release 5-HT from neurons. However, the effects of these drugs on other monoamine transporters are not well characterized. In this study, we examined the interaction of stereoisomers of (+/-)-fenfluramine and (+/-)-norfenfluramine with transporters for 5-HT, norepinephrine (NE), and dopamine (DA). Results from in vitro assays confirmed these drugs are potent substrates for 5-HT transporters: (+) fenfluramine, (-)-fenfluramine, (+)-norfenfluramine, and (-)-norfenfluramine released [3H]5-HT from synaptosomes with EC50 values of 52, 147, 59, and 287 nM, respectively. Importantly, (+)-fenfluramine and (+)-norfenfluramine released [3H]NE with EC50 values of 302 and 73 nM. Results from in vivo microdialysis experiments showed that intravenous injection of (+)-norfenfluramine elevates extracellular levels of 5-HT, NE, and DA in rat frontal cortex. The effects of (+)-norfenfluramine on NE and DA were antagonized by pretreatment with the NE uptake blocker nisoxetine. In summary, administration of fenfluramines can increase synaptic levels of 5-HT, NE, and DA in the cortex, and (+) norfenfluramine likely contributes to these effects. Release of NE and DA evoked by (+)-norfenfluramine is at least partly mediated via NE transporters. Our results emphasize the potential involvement of noradrenergic mechanisms in the actions of fenfluramines. PMID- 12649308 TI - Kinetics of NAD(P)H:quinone oxidoreductase I (NQO1) inhibition by mitomycin C in vitro and in vivo. AB - The bioreductive activation of the antitumor quinone mitomycin C (MMC) by NAD(P)H: quinone oxidoreductase 1 (NQO1) is complicated by the ability of MMC to also act as a mechanism-based inhibitor of NQO1 in a pH dependent manner. Inhibition of NQO1 by MMC has been studied in purified enzyme preparations and in cultured cells but has not determined in vivo. In the studies presented here, NQO1 activity was measured in mouse tissues following treatment with MMC or the potent mechanism-based human NQO1 inhibitor 5-methoxy-1,2-dimethyl-[(4 nitrophenoxy)methyl]indole-4,7-dione (ES936). NQO1 activity was significantly decreased at 1, 2, and 4 h following MMC (10 or 20 mg/kg) treatment in kidney and lung but was unchanged in brain, heart, liver, and bladder. ES936 (1 mg/kg) treatment led to a significant and much more potent inhibition of NQO1 in all murine tissues analyzed except for bladder. To extrapolate these in vivo results from mice to humans, the species-specific kinetics of NQO1 inactivation by MMC was determined in vitro using mouse, rat, and human recombinant NQO1 proteins. Results showed the inactivation kinetics of mouse and human proteins by MMC were similar. Treatment of human and murine endothelial cells with MMC or ES936 showed similar inhibition of NQO1 activity. The aforementioned results clearly demonstrate that MMC can serve as a substrate for NQO1 in vivo; however, the metabolism resulting in enzyme inactivation is possibly tissue-specific. Furthermore, the kinetic similarities for inactivation between murine and human forms of NQO1 show these results are apropos to clinical use of MMC. PMID- 12649309 TI - Fractal activity generated independently by medullary sympathetic premotor and preganglionic sympathetic neurons. AB - In anesthetized cats with cervical spinal cord transection, Fano factor analysis was used to test for time-scale invariant (fractal) fluctuations in spike counts of single preganglionic cervical sympathetic neurons (PSNs) and putative sympathetic premotor neurons located in the rostral ventrolateral medulla (RVLM) and caudal medullary raphe. The medullary neurons exhibited cardiac-related activity, and their axons projected to the spinal cord, as demonstrated by antidromic activation. The variance-to-mean spike count ratio (Fano factor) was plotted as a function of the window size used to count spikes. The Fano factor curves for seven PSNs, eight RVLM neurons, and eight raphe neurons contained a power law relationship extending over more than one time scale. In these cases, random shuffling of the interspike intervals in the original time series eliminated the power law relationship. Thus the power law relationships can be attributed to long-range correlations among interspike intervals rather than simply to the distribution of the intervals that is not changed by shuffling the data. It is concluded that PSNs and sympathetic premotor neurons in the medulla can independently generate fractal firing patterns. PMID- 12649310 TI - Ionic mechanisms mediating oscillatory membrane potentials in wide-field retinal amacrine cells. AB - Particular types of amacrine cells of the vertebrate retina show oscillatory membrane potentials (OMPs) in response to light stimulation. Historically it has been thought the oscillations arose as a result of circuit properties. In a previous study we found that in some amacrine cells, the ability to oscillate was an intrinsic property of the cell. Here we characterized the ionic mechanisms responsible for the oscillations in wide-field amacrine cells (WFACs) in an effort to better understand the functional properties of the cell. The OMPs were found to be calcium (Ca2+) dependent; blocking voltage-gated Ca2+ channels eliminated the oscillations, whereas elevating extracellular Ca2+ enhanced them. Strong intracellular Ca2+ buffering (10 mM EGTA or bis-(o-aminophenoxy)-N,N,N',N' tetraacetic acid) eliminated any attenuation in the OMPs as well as a Ca2+ dependent inactivation of the voltage-gated Ca2+ channels. Pharmacological and immunohistochemical characterization revealed that WFACs express L- and N-type voltage-sensitive Ca2+ channels. Block of the L-type channels eliminated the OMPs, but omega-conotoxin GVIA did not, suggesting a different function for the N type channels. The L-type channels in WFACs are functionally coupled to a set of calcium-dependent potassium (K(Ca)) channels to mediate OMPs. The initiation of OMPs depended on penitrem-A-sensitive (BK) K(Ca) channels, whereas their duration is under apamin-sensitive (SK) K(Ca) channel control. The Ca2+ current is essential to evoke the OMPs and triggering the K(Ca) currents, which here act as resonant currents, enhances the resonance as an amplifying current, influences the filtering characteristics of the cell membrane, and attenuates the OMPs via CDI of the L-type Ca2+ channel. PMID- 12649311 TI - Dorsal and ventral distribution of excitable and synaptic properties of neurons of the bed nucleus of the stria terminalis. AB - The bed nucleus of the stria terminalis (BNST) is a structure uniquely positioned to integrate stress information and regulate both stress and reward systems. Consistent with this arrangement, evidence suggests that the BNST, and in particular the noradrenergic input to this structure, is a key component of affective responses to drugs of abuse. We have utilized an in vitro slice preparation from adult mice to determine synaptic and membrane properties of these cells, focusing on the dorsal and ventral subdivisions of the anterolateral BNST (dBNST and vBNST) because of the differential noradrenergic input to these two regions. We find that while resting membrane potential and input resistance are comparable between these subdivisions, excitable properties, including a low threshold spike (LTS) likely mediated by T-type calcium channels and an Ih dependent potential, are differentially distributed. Inhibitory and excitatory postsynaptic potentials (IPSPs and EPSPs, respectively) are readily evoked in both dBNST and vBNST. The fast IPSP is predominantly GABAA-receptor mediated and is partially blocked by the AMPA/kainate-receptor antagonist CNQX. In the presence of the GABAA-receptor antagonist picrotoxin, cells in dBNST but not vBNST are more depolarized and have a higher input resistance, suggesting tonic GABAergic inhibition of these cells. The EPSPs elicited in BNST are monosynaptic, exhibit paired pulse facilitation, and contain both an AMPA- and an N-methyl-d aspartate (NMDA) receptor-mediated component. These data support the hypothesis that neurons of the dorsal and ventral BNST differentially integrate synaptic input, which is likely of behavioral significance. The data also suggest mechanisms by which information may flow through stress and reward circuits. PMID- 12649312 TI - Adaptation and temporal decorrelation by single neurons in the primary visual cortex. AB - Limiting redundancy in the real-world sensory inputs is of obvious benefit for efficient neural coding, but little is known about how this may be accomplished by biophysical neural mechanisms. One possible cellular mechanism is through adaptation to relatively constant inputs. Recent investigations in primary visual (V1) cortical neurons have demonstrated that adaptation to prolonged changes in stimulus contrast is mediated in part through intrinsic ionic currents, a Ca2+ activated K+ current (IKCa) and especially a Na+-activated K+ current (IKNa). The present study was designed to test the hypothesis that the activation of adaptation ionic currents may provide a cellular mechanism for temporal decorrelation in V1. A conductance-based neuron model was simulated, which included an IKCa and an IKNa. We show that the model neuron reproduces the adaptive behavior of V1 neurons in response to high contrast inputs. When the stimulus is stochastic with 1/f 2 or 1/f-type temporal correlations, these autocorrelations are greatly reduced in the output spike train of the model neuron. The IKCa is effective at reducing positive temporal correlations at approximately 100-ms time scale, while a slower adaptation mediated by IKNa is effective in reducing temporal correlations over the range of 1-20 s. Intracellular injection of stochastic currents into layer 2/3 and 4 (pyramidal and stellate) neurons in ferret primary visual cortical slices revealed neuronal responses that exhibited temporal decorrelation in similarity with the model. Enhancing the slow afterhyperpolarization resulted in a strengthening of the decorrelation effect. These results demonstrate the intrinsic membrane properties of neocortical neurons provide a mechanism for decorrelation of sensory inputs. PMID- 12649314 TI - Modulation of striatal single units by expected reward: a spiny neuron model displaying dopamine-induced bistability. AB - Single-unit activity in the neostriatum of awake monkeys shows a marked dependence on expected reward. Responses to visual cues differ when animals expect primary reinforcements, such as juice rewards, in comparison to secondary reinforcements, such as tones. The mechanism of this reward-dependent modulation has not been established experimentally. To assess the hypothesis that direct neuromodulatory effects of dopamine on spiny neurons can account for this modulation, we develop a computational model based on simplified representations of key ionic currents and their modulation by D1 dopamine receptor activation. This minimal model can be analyzed in detail. We find that D1-mediated increases of inward rectifying potassium and L-type calcium currents cause a bifurcation: the native up/down state behavior of the spiny neuron model becomes truly bistable, which modulates the peak firing rate and the duration of the up state and introduces a dependence of the response on the past state history. These generic consequences of dopamine neuromodulation through bistability can account for both reward-dependent enhancement and suppression of spiny neuron single-unit responses to visual cues. We validate the model by simulating responses to visual targets in a memory-guided saccade task; our results are in close agreement with the main features of the experimental data. Our model provides a conceptual framework for understanding the functional significance of the short-term neuromodulatory actions of dopamine on signal processing in the striatum. PMID- 12649315 TI - Effects of attention and emotion on repetition priming and their modulation by cholinergic enhancement. AB - We examined whether behavioral and neural effects of repeating faces are modulated by independent factors of selective attention, emotion, and cholinergic enhancement, during functional MRI. Face repetition occurred either between task relevant (spatially attended) or task-irrelevant (unattended) stimuli; faces could be fearful or neutral; subjects received either placebo or physostigmine. Under placebo, a reaction time advantage occurred with repetition (i.e., priming) that did not differ between levels of attention, but was attenuated with emotion. Inferior temporo-occipital cortex demonstrated repetition decreases to both attended and unattended faces, and showed either equivalent or greater repetition decreases with emotional compared with neutral faces. By contrast, repetition decreases were attenuated for emotional relative to neutral faces in lateral orbitofrontal cortex. These results distinguish automatic repetition effects in sensory cortical regions from repetition effects modulated by emotion in orbitofrontal cortex, which parallel behavioral effects. Under physostigmine, unlike placebo, behavioral repetition effects were seen selectively for attended faces only, whereas emotional faces no longer impaired priming. Physostigmine enhanced repetition decreases in inferior occipital cortex selectively for attended faces, and reversed the emotional interaction with repetition in lateral orbitofrontal cortex. Thus we show that cholinergic enhancement both augments a neural signature of priming and modulates the effects of attention and emotion on behavioral and neural consequences of repetition. PMID- 12649316 TI - Effect of semicircular canal stimulation on the perception of the visual vertical. AB - The subjective visual vertical (SVV) is usually considered a measure of otolith function. Herewith we investigate the influence of semicircular canal (SCC) stimulation on the SVV by rotating normal subjects in yaw about an earth-vertical axis, with velocity steps of +/- 90 degrees /s, for 60 s. SVV was assessed by setting an illuminated line to perceived earth vertical in darkness, during a per and postrotary period. Four head positions were tested: upright, 30 degrees backward (chin up) or forward, and approximately 40 degrees forward from upright. During head upright/backward conditions, a significant SVV tilt (P < 0.01) in the direction opposite to rotation was found that reversed during postrotary responses. The rotationally induced SVV tilt had a time constant of decay of approximately 30 s. Rotation with the head 30 degrees forward did not affect SVV, whereas the 40 degrees forward tilt caused a direction reversal of SVV responses compared with head upright/backward. Spearman correlation values (Rho) between individual SCC efficiencies in different head positions and mean SVV tilts were 0.79 for posterior, 0.34 for anterior, and - 0.80 for horizontal SCCs. Three dimensional video-oculography showed that SVV and torsional eye position measurements were highly correlated (0.83) and in the direction opposite to the slow phase torsional vestibuloocular reflex. IN CONCLUSION: 1) during yaw axis rotation without reorientation of the head with respect to gravity, the SVV is influenced by SCC stimulation; 2) this effect is mediated by the vertical SCCs, particularly the posterior SCCs; 3) rotationally induced SVV changes are due to torsional ocular tilt; 4) SVV and ocular tilts occur in the "anticompensatory," fast phase direction of the torsional nystagmus; and 5) clinically, abnormal SVV tilts cannot be considered a specific indication of otolith system dysfunction. PMID- 12649317 TI - Long-term plasticity of ipsilesional medial vestibular nucleus neurons after unilateral labyrinthectomy. AB - Unilateral labyrinthectomy results in oculomotor and postural disturbances that regress in a few days during vestibular compensation. The long-term (after 1 mo) consequences of unilateral labyrinthectomy were investigated by characterizing the static and dynamic membrane properties of the ipsilesional vestibular neurons recorded intracellularly in guinea pig brain stem slices. We compared the responses of type A and type B medial vestibular nucleus neurons identified in vitro to current steps and ramps and to sinusoidal currents of various frequencies. All ipsilesional vestibular neurons were depolarized by 6-10 mV at rest compared with the cells recorded from control slices. Both their average membrane potential and firing threshold were more depolarized, which suggests that changes in active conductances compensated for the loss of excitatory afferents. The afterhyperpolarization and discharge regularity of type B but not type A neurons were increased. All ipsilesional vestibular cells became more sensitive to current injections over a large range of frequencies (0.2-30 Hz), but this increase in sensitivity was greater for type B than for type A neurons. This was associated with an increase of the peak frequency of linear response restricted to type B neurons, from 4-6 to 12-14 Hz. Altogether, we show that long term vestibular compensation involves major changes in the membrane properties of vestibular neurons on the deafferented side. Many of the static and dynamic membrane properties of type B neurons became more similar to those of type A neurons than in control slices, leading to an increase in the overall homogeneity of medial vestibular nucleus neurons. PMID- 12649318 TI - Functional significance of cannabinoid-mediated, depolarization-induced suppression of inhibition (DSI) in the hippocampus. AB - A number of recent studies have demonstrated that a well-known form of short-term plasticity at hippocampal GABAergic synapses, called depolarization-induced suppression of inhibition (DSI), is in fact mediated by the retrograde actions of endocannabinoids released in response to depolarization of the postsynaptic cells. These studies suggest that endogenous cannabinoids may play an important role in regulating inhibitory tone in the mammalian CNS. Despite the widespread interest and potential physiological importance of DSI, many questions regarding the physiological relevance of DSI remain. To that end, this study set out to define the specific limiting conditions that could elicit DSI at GABAergic synapses in CA1 hippocampal pyramidal neurons and to determine if DSI could be elicited with pulse trains that mimic hippocampal cell-firing patterns that occur in vivo. Whole cell recordings from hippocampal neurons under voltage-clamp configuration were made in rat hippocampal slices. Spontaneous and evoked gamma aminobutyric acid-A (GABAA) receptor-mediated inhibitory postsynaptic currents (sIPSCs and eIPSCs, respectively) were recorded prior to and following depolarization of CA1 hippocampal pyramidal cells. Depolarizing voltage pulses were shaped to evoke currents in QX-314-treated cells similar to those accompanying single spontaneous voltage-clamped action potentials recorded from the soma. Attempts were made to elicit DSI with trains of these pulses that mimicked hippocampal cell firing patterns in vivo, for instance, when animals traverse place fields or are performing a short-term memory task. DSI could not be elicited by such pulse trains or by a number of other combinations of behaviorally specific firing parameters. The minimum duration of depolarization necessary to elicit DSI in hippocampal neurons determined by paired-pulse manipulation was 50 -75 ms at a critical interval of 20 -30 ms between pulse pairs. Under the conditions tested, the normal firing patterns of hippocampal neurons that occur in vivo do not appear to elicit DSI. PMID- 12649319 TI - Neuregulin 1-erbB2 signaling is required for the establishment of radial glia and their transformation into astrocytes in cerebral cortex. AB - Radial glial cells and astrocytes function to support the construction and maintenance, respectively, of the cerebral cortex. However, the mechanisms that determine how radial glial cells are established, maintained, and transformed into astrocytes in the cerebral cortex are not well understood. Here, we show that neuregulin-1 (NRG-1) exerts a critical role in the establishment of radial glial cells. Radial glial cell generation is significantly impaired in NRG mutants, and this defect can be rescued by exogenous NRG-1. Down-regulation of expression and activity of erbB2, a member of the NRG-1 receptor complex, leads to the transformation of radial glial cells into astrocytes. Reintroduction of erbB2 transforms astrocytes into radial glia. The activated form of the Notch1 receptor, which promotes the radial glial phenotype, activates the erbB2 promoter in radial glial cells. These results suggest that developmental changes in NRG-1 erbB2 interactions modulate the establishment of radial glia and contribute to their appropriate transformation into astrocytes. PMID- 12649320 TI - Processive DNA synthesis observed in a polymerase crystal suggests a mechanism for the prevention of frameshift mutations. AB - DNA polymerases replicate DNA by adding nucleotides to a growing primer strand while avoiding frameshift and point mutations. Here we present a series of up to six successive replication events that were obtained by extension of a primed template directly in a crystal of the thermostable Bacillus DNA polymerase I. The 6-bp extension involves a 20-A translocation of the DNA duplex, representing the largest molecular movement observed in a protein crystal. In addition, we obtained the structure of a "closed" conformation of the enzyme with a bound triphosphate juxtaposed to a template and a dideoxy-terminated primer by constructing a point mutant that destroys a crystal lattice contact stabilizing the wild-type polymerase in an "open" conformation. Together, these observations allow many of the steps involved in DNA replication to be observed in the same enzyme at near atomic detail. The successive replication events observed directly by catalysis in the crystal confirm the general reaction sequence deduced from observations obtained by using several other polymerases and further refine critical aspects of the known reaction mechanism, and also allow us to propose new features that concern the regulated transfer of the template strand between a preinsertion site and an insertion site. We propose that such regulated transfer is an important element in the prevention of frameshift mutations in high fidelity DNA polymerases. The ability to observe processive, high-fidelity replication directly in a crystal establishes this polymerase as a powerful model system for mechanistic studies in which the structural consequences of mismatches and DNA adducts are observed. PMID- 12649321 TI - A model of the glycine receptor deduced from Brownian dynamics studies. AB - We have developed a three-dimensional model of the alpha1 homomeric glycine receptor by using Brownian dynamics simulations to account for its observed physiological properties. The model channel contains a large external vestibule and a shallow internal vestibule, connected by a narrow, cylindrical selectivity filter. Three rings of charged residues from the pore-lining M2 domain are modeled as point charges in the protein. Our simulations reproduce many of the key features of the channel, such as the current-voltage profiles, permeability ratios, and ion selectivity. When we replace the ring of alanine residues lining the selectivity filter with glutamates, the mutant model channel becomes permeable to cations, as observed experimentally. In this mutation, anions act as chaperones for sodium ions in the extracellular vestibule, and together they penetrate deep inside the channel against a steep energy barrier encountered by unaccompanied ions. Two subsequent amino acid mutations increase the cation permeability, enabling monovalent cations to permeate through the channel unaided and divalent cations to permeate when chaperoned by anions. These results illustrate the key structural features and underlying mechanism for charge selectivity in the glycine receptor. PMID- 12649322 TI - Calpain regulates neutrophil chemotaxis. AB - Cell polarization is required for directed cell migration. We investigated the role of the calcium-dependent protease calpain during neutrophil chemotaxis and found that calpain inhibition induced neutrophil adhesion, polarization, and rapid chemokinesis in the absence of exogenous activators. Resting neutrophils display constitutive calpain activity with mu-calpain being the predominant active isoform. Our findings suggest that constitutive calpain activity in resting neutrophils may function as a negative regulator of protrusion and migration. Specific inhibition of mu-calpain, but not m-calpain, induced neutrophil polarization and chemokinesis. In contrast to IL-8-induced chemokinesis, the chemokinesis induced by calpain inhibition was not reduced in the presence of pertussis toxin, suggesting that calpain functions downstream of G protein-coupled receptors. Further, both calpain inhibition and stimulation with IL-8 and formyl-Met-Leu-Phe (fMLP) induced an increase in Cdc42 and Rac activation. These findings are consistent with the involvement of calpain in chemotaxis pathways. Accordingly, calpain inhibition decreased neutrophil chemotaxis and directional persistence in a gradient of IL-8 and fMLP. Together, these data reveal a previously uncharacterized function for calpain in neutrophils and suggest that localized modulation of calpain activity may regulate neutrophil chemotaxis downstream of G-protein-coupled receptors. PMID- 12649323 TI - Nutritional supplement chromium picolinate causes sterility and lethal mutations in Drosophila melanogaster. AB - The nutritional dietary supplement chromium picolinate, [Cr(pic)(3)], has gained much notoriety as a safe supplement that supposedly promotes fat loss and muscle enhancement in humans. Thus, a significant industry has materialized around the incorporation of [Cr(pic)(3)] in many sports foods and drinks and a variety of weight loss products. However, in vitro studies have suggested that low levels of [Cr(pic)(3)] in the presence of biological reducing agents can catalytically generate reactive oxygen species, and recent in vivo studies have detected oxidative damage in rats receiving the supplement. The potential deleterious in vivo effects of this activity were examined by using Drosophila melanogaster. [Cr(pic)(3)], but not CrCl(3), at levels of 260 microg Crkg food or less were found to lower the success rate of pupation and eclosion and to arrest development of pupae in a concentration dependent fashion. X-linked lethal analysis indicates that the supplement greatly enhances the rate of appearance of lethal mutations and dominant female sterility. PMID- 12649324 TI - Synthetic modeling of zinc thiolates: quantitative assessment of hydrogen bonding in modulating sulfur alkylation rates. AB - A series of mononuclear zinc thiolate complexes have been prepared and fully characterized. The reactions of the complexes with alkyl halides, leading to zinc halides and the corresponding thioethers, have been examined by kinetic methods. In toluene, the reactions obey a second-order rate law displaying activation parameters consistent with a S(N)2 attack of the zinc-bound thiolate on the carbon electrophile. Intramolecular hydrogen bonding of an amide NH to the thiolate sulfur reduces the nucleophilicity and consequently, the rate of alkylation more than 30-fold at 25 degrees C. The H-bonding shows an inverse HD isotope effect of 0.33 (60 degrees C) ascribed to differential H-bonding for the two isotopomers due to zero point energy differences. These model studies provide quantitative evaluation of H-bonding on reaction rates relevant to zinc thiol activating proteins. PMID- 12649325 TI - Global and specific transcriptional repression by the histone H3 amino terminus in yeast. AB - The yeast CHA1 promoter is activated in the presence of serine or threonine. Activation requires the Cha4p activator, and it results in perturbation of a nucleosome that incorporates the TATA element under noninducing conditions. We show that in yeast lacking the amino terminus of histone H3, the promoter is constitutively active and the chromatin is concomitantly perturbed. This derepression occurs in the absence of elevated intracellular levels of serine or threonine and is not observed in cells lacking Rpd3p, Tup1p, or the amino terminus of histone H4. Furthermore, derepression in the absence of the H3 amino terminus requires the primary activator of this promoter, Cha4p, which we show by chromatin immunoprecipitation to be constitutively bound to the CHA1 promoter in WT yeast. Thus, the H3 amino terminus is required to prevent Cha4p from activating CHA1 in the absence of inducer. We also present results of a microarray experiment showing that the H3 amino terminus has a substantial repressive effect on a genome-wide scale. PMID- 12649326 TI - Importance of the CNGA4 channel gene for odor discrimination and adaptation in behaving mice. AB - Odor stimulation of olfactory sensory neurons (OSNs) leads to both the activation and subsequent desensitization of a heteromultimeric cyclic-nucleotide-gated (CNG) channel present in these cells. The native olfactory CNG channel consists of three distinct subunits: CNGA2, CNGA4, and CNGB1b. Mice in which the CNGA4 gene has been deleted display defective Ca(2+)calmodulin-dependent inhibition of the CNG channel, resulting in a striking reduction in adaptation of the odor induced electrophysiological response in the OSNs. These mutants therefore afford an excellent opportunity to assess the importance of Ca(2+)-mediated CNG channel desensitization for odor discrimination and adaptation in behaving animals. By using an operant conditioning paradigm, we show that CNGA4-null mice are profoundly impaired in the detection and discrimination of olfactory stimuli in the presence of an adapting background odor. The extent of this impairment depends on both the concentration and the molecular identity of the adapting stimulus. Thus, Ca(2+)-dependent desensitization of the odor response in the OSNs mediated by the CNGA4 subunit is essential for normal odor sensation and adaptation of freely behaving mice, preventing saturation of the olfactory signal transduction machinery and extending the range of odor detection and discrimination. PMID- 12649327 TI - Antisense protein tyrosine phosphatase 1B reverses activation of p38 mitogen activated protein kinase in liver of ob/ob mice. AB - Phosphorylation of stress-activated kinase p38, a MAPK family member, was increased in liver of ob/ob diabetic mice relative to lean littermates. Treatment of ob/ob mice with protein tyrosine phosphatase 1B (PTP1B) antisense oligonucleotides (ASO) reduced phosphorylation of p38 in liver-to below lean littermate levels-and normalized plasma glucose while reducing plasma insulin. Phosphorylation of ERK, but not JNK, was also decreased in ASO-treated mice. PTP1B ASO decreased TNFalpha protein levels and phosphorylation of the transcription factor cAMP response element binding protein (CREB) in liver, both of which can occur through decreased phosphorylation of p38 and both of which have been implicated in insulin resistance or hyperglycemia. Decreased p38 phosphorylation was not directly due to decreased phosphorylation of the kinases that normally phosphorylate p38-MKK3 and MKK6. Additionally, p38 phosphorylation was not enhanced in liver upon insulin stimulation of ASO-treated ob/ob mice (despite increased activation of other signaling molecules) corroborating that p38 is not directly affected via the insulin receptor. Instead, decreased phosphorylation of p38 may be due to increased expression of MAPK phosphatases, particularly the p38/ERK phosphatase PAC1 (phosphatase of activated cells). This study demonstrates that reduction of PTP1B protein using ASO reduces activation of p38 and its substrates TNFalpha and CREB in liver of diabetic mice, which correlates with decreased hyperglycemia and hyperinsulinemia. PMID- 12649328 TI - Phosphodiesterase regulation is critical for the differentiation and pattern of gene expression in granulosa cells of the ovarian follicle. AB - Feedback regulations are integral components of the cAMP signaling required for most cellular processes, including gene expression and cell differentiation. Here, we provide evidence that one of these feedback regulations involving the cyclic nucleotide phosphodiesterase PDE4D plays a critical role in cAMP signaling during the differentiation of granulosa cells of the ovarian follicle. Gonadotropins induce PDE4D mRNA and increase the cAMP hydrolyzing activity in granulosa cells, demonstrating that a feedback regulation of cAMP is operating in granulosa cells in vivo. Inactivation of the PDE4D by homologous recombination is associated with an altered pattern of cAMP accumulation induced by the gonadotropin LH/human chorionic gonadotropin (hCG), impaired female fertility, and a markedly decreased ovulation rate. In spite of a disruption of the cAMP response, LH/hCG induced P450 side chain cleavage expression and steroidogenesis in a manner similar to wild-type controls. Morphological examination of the ovary of PDE4D-/- mice indicated luteinization of antral follicles with entrapped oocytes. Consistent with the morphological finding of unruptured follicles, LH/hCG induction of genes involved in ovulation, including cyclooxygenase-2, progesterone receptor, and the downstream genes, is markedly decreased in the PDE4D-/- ovaries. These data demonstrate that PDE4D regulation plays a critical role in gonadotropin mechanism of action and suggest that the intensity and duration of the cAMP signal defines the pattern of gene expression during the differentiation of granulosa cells. PMID- 12649329 TI - Cyclin D2 compensates for the loss of cyclin D1 in estrogen-induced mouse uterine epithelial cell proliferation. AB - The cell cycle-regulatory protein, cyclin D1, is the sensor that connects the intracellular cell cycle machinery to external signals. Given this central role in the control of cell proliferation, it was surprising that mice lacking the cyclin D1 gene were viable and fertile. Fertility requires 17beta-estradiol (E2) induced uterine luminal epithelial cell proliferation. In these cells E2 causes the translocation of cyclin D1/cyclin-dependent kinase 4 (CDK4) from the cytoplasm into the nucleus with the consequent phosphorylation of the retinoblastoma protein. In cyclin D1 null mice, E2 also induces retinoblastoma protein phosphorylation and DNA synthesis in a normal manner. CDK4 activity was slightly reduced in the D1 null mice compared with wild-type mice. This CDK4 activity was due to complexes of cyclin D2/CDK4. Cyclin D2 was translocated into the nucleus in response to E2 in the cyclin D1-/- mice to a much greater degree than in wild-type mice. This cyclin D2/CDK4 complex was also able to bind p27kip1 in cyclin D1-/- uterine luminal epithelial cells, allowing for the activation of CDK2. Our data show that in vivo cyclin D2 can completely compensate for the loss of cyclin D1 and reinforces the conclusions that cyclin Ds are the central regulatory point in the proliferative responses of epithelial cells to estrogens. PMID- 12649331 TI - Lack of SYT-SSX fusion transcripts in malignant peripheral nerve sheath tumors on RT-PCR analysis of 34 archival cases, by Tamborini E., et al. PMID- 12649332 TI - DNA arrays in clinical oncology: promises and challenges. AB - Cancer is a complex genetic disease characterized by the accumulation of multiple molecular alterations. Current diagnostic and prognostic classifications, based on clinical and pathologic factors, are insufficient to reflect the whole clinical heterogeneity of tumors. Most current anticancer agents do not differentiate between cancerous and normal cells, leading sometimes to disastrous adverse effects. Recent advances in human genome research and high-throughput molecular technologies make it possible finally to tackle the molecular complexity of malignant tumors. With DNA array technology, mRNA expression levels of thousands of genes can be measured simultaneously in a single assay. Oncology is benefiting on multiple fronts. Gene expression profiles are revealing new biologically and clinically relevant tumor subclasses previously indistinguishable and are identifying new diagnostic and prognostic biomarkers as well as new potential therapeutic targets. Here, we review the technology and present clinical applications for which promising results have been obtained. Finally, we discuss issues that must be resolved in the near future to allow DNA arrays to translate into benefits for cancer patients. PMID- 12649333 TI - CD36/fatty acid translocase in rats: distribution, isolation from hepatocytes, and comparison with the scavenger receptor SR-B1. AB - The new mAb UA009 recognizes an antigen expressed by microvascular endothelium, by lymphatic endothelium, and by some epithelia in a number of organs, including the small intestine, lactating mammary gland, kidney, lung, sebaceous glands, and circumvallate papillae of the tongue. This antigen is also expressed abundantly in the splenic red pulp and marginal zone and by monocytes, macrophages, and erythrocytes (but not by platelets). Among tissues that store or metabolize fatty acids, the antigen is expressed by adipocytes, cardiomyocytes, and red skeletal muscle. Importantly, it is expressed by steroidogenic cells in the adrenal gland, testis, and ovary, whereas in the liver it is expressed by hepatocytes in a pattern that is dependent on gender and genetic background. mAb UA009 immunoprecipitated a mol wt 85-kDa surface protein from detergent extracts of hepatocytes from Dark Agouti female rats. The N-terminal amino acid sequence of this protein was identical to fatty acid translocase (FAT), the rat cluster of differentiation 36 (CD36) ortholog. The mAb also reacted with COS-7 cells transfected with cDNA encoding FAT. cDNAs encoding a CD36/FAT-like polypeptide were prepared from both liver and heart RNA by RT-PCR. The nucleotide sequences obtained from these cDNAs (Dark Agouti rats) revealed identity and 99% similarity, respectively, with the published sequences of Cd36/Fat in rats of the Wistar and Sprague-Dawley strains. The absence of the UA009 antigen in CD36/FAT deficient SHR/N rats confirmed the identity of the UA009 antigen and CD36/FAT. We suggest that CD36/FAT might function in the liver as a sex-regulated accessory molecule, either in reverse cholesterol transport and/or in fatty acid uptake. PMID- 12649334 TI - Cyclooxygenase-2-dependent and thromboxane-dependent vascular and bronchial responses are regulated via p38 mitogen-activated protein kinase in control and endotoxin-primed rat lungs. AB - Mitogen-activated protein kinases (MAPKs) are part of an intracellular signaling machinery consisting of three known distinct pathways, each leading to activation of a different protein kinase: p38, ERK (extracellular signal-regulated kinase), or JNK (c-Jun N-terminal kinase). We investigated the role of the p38 MAPK pathway in the phenomenon of lung endotoxin "priming": incubation of perfused rat lungs with lipopolysaccharide (LPS) for 2 hours results in drastically enhanced cyclooxygenase-2-dependent and thromboxane synthase-dependent vasoconstriction and bronchoconstriction, including edema formation in response to a second inflammatory stimulus, such as arachidonic acid application. Two unrelated selective inhibitors of p38 (SB203580 and SC-68376) dose dependently suppressed the arachidonic acid-induced pulmonary artery pressor response, edema formation, and bronchoconstrictor response in both control lungs and lungs that underwent preceding endotoxin priming. In parallel, thromboxane, but not prostacyclin, released into the lung perfusate was dose dependently inhibited. Using immunohistochemical techniques in combination with quantitative microdensitometry, p38 was detected in nearly all cell types in control lungs, whereas the activated form p-p38 was only expressed in certain cell types, eg, bronchial epithelial cells, endothelial cells, alveolar macrophages, and vascular smooth muscle cells (SMC) of small vessels. In response to endotoxin, p-p38 expression was additionally observed in septal cells, bronchial SMC, and vascular SMC of larger pulmonary vessels and was increased in most other cell types including small-vessel SMC. We conclude that both immunolocalization of p38 activity and pharmacologic interventions support a strong role of the p38 MAPK pathway in establishing an active cyclooxygenase-2/thromboxane synthase axis in vascular and bronchial SMC, with up-regulation of this signaling cascade occurring in LPS priming and being responsible for enhanced pulmonary artery pressor response, edema formation, and bronchoconstriction. Moreover, LPS induces or increases phosphorylation of p38 in other lung cell types. The physiologic consequences of these events remain to be established. PMID- 12649335 TI - Oxysterol 7alpha-hydroxylase (CYP39A1) in the ciliary nonpigmented epithelium of bovine eye. AB - The CYP39A1 oxysterol 7alpha-hydroxylase preferentially catalyzes the 7alpha hydroxylation of 24-hydroxycholesterol and has been suggested to play a role in the alternative bile acid synthesis pathway in the liver. The presence of CYP39A1 oxysterol 7alpha-hydroxylase has been reported only in the liver. To investigate the physiologic characteristics of the ciliary processes in bovine ocular tissues, we raised a mAb, 42C, against nonpigmented epithelial (NPE) cells, which have tight junctions that act as a blood-aqueous barrier and are involved in producing aqueous humor and maintaining ocular homeostasis. Immunohistochemical analysis showed that 42C antibody reacted intensely with an antigen in the NPE cells of the ciliary processes but not with other ocular tissues. The SDS-PAGE profile of immunoaffinity-purified antigens from bovine ciliary processes showed a predominant protein of molecular mass of 44.0 kDa. The amino acid sequence of this antigenic protein was identical to human CYP39A1 oxysterol 7alpha hydroxylase. Immunoreactivity with 42C antibody was found only in hepatocytes and ocular tissues. These data suggest a new physiologic function for the CYP39A1 oxysterol 7alpha-hydroxylase in addition to the production of bile acids and provide new insight into the physiologic role of the ciliary NPE cells concerning the metabolism of sterols in the eye. PMID- 12649336 TI - Attenuated apoptosis of B cell activating factor-expressing cells in primary Sjogren's syndrome. AB - B cell activating factor (BAFF) is known to be a powerful regulator of B-cell differentiation and proliferation. The aim of this study was to assess the incidence of apoptosis among BAFF-expressing cells in Sjogren's syndrome (SS) salivary gland tissue. We performed double stainings of BAFF together with one of the markers, CD21, CD68, CD40, Fas, Bcl-2 or Bax, and monitored apoptosis among BAFF expressing cells by using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-digoxigenin nick-end labeling method. A significantly lower level of apoptosis among the BAFF-expressing cells was detected in salivary glands from patients with SS compared with controls (p = 0.03). Furthermore, no difference in the coexpression of Fas or CD40 together with BAFF was detected between patients and controls. Coexpression of the pro apoptotic molecule Bax together with BAFF was nonsignificantly decreased in patients with SS compared with controls. Our results suggest that a reduced level of apoptosis among BAFF expressing cells might lead to longer-existing BAFF expression within these cells and thereby maintain signaling for tissue-infiltrating B cells to proliferate and mature. PMID- 12649337 TI - Time-dependent interactions of oxidant-sensitive fluoroprobes with inhibitors of cellular metabolism. AB - SUMMARY: We tested three oxidant sensitive fluoroprobes (dihydrorhodamine [DHR], 2',7'-dichlorodihydrofluorescein [H(2)DCF], and dihydroethidium [DHE]) for interactions with three inhibitors of mitochondrial electron transport. DHR, H(2)DCF, and DHE produced large time-dependent increases in fluorescence in a cell-free medium that contained either of the site III inhibitors antimycin (A) and 2-heptyl-4-hydroxy-quinoline-N-oxide but minimal increases in medium that contained another site III inhibitor, myxothiazol (Mx). The interactions between A and each of the fluoroprobes occurred at concentrations of agent/probe that are frequently used in experiments designed to investigate cellular oxidant production. To define more effectively the nature of these agent/probe interactions, we determined the oxygen dependence of the interactions between A and each probe. The A/H(2)DCF and A/DHR interactions either were highly oxygen dependent or exhibited a small degree of oxygen dependence, respectively, whereas the A/DHE interaction was oxygen-independent. Finally, we determined multiple ways to reduce the impact of the agent/probe interaction on data acquisition. The addition of either fetal bovine serum (10%) or albumin (5%) to the media abolished the A/DHR and A/H(2)DCF interactions. Shifting the excitation wavelength of DHE (from 470 to 530 nm) reduced measurement of the A/DHE interaction while preserving measurement of the intracellular signal. Collectively, these results emphasize the importance of testing for interactions between agents and probes, because these interactions can interfere with the accurate interpretation of experimental results. In addition, the methods presented for circumventing these interactions may be applicable to other experiments in which agent/probe interactions are an obstacle to accurate interpretation of the experimental results. PMID- 12649338 TI - Galectin-1 accumulation in the ovary carcinoma peritumoral stroma is induced by ovary carcinoma cells and affects both cancer cell proliferation and adhesion to laminin-1 and fibronectin. AB - Galectin-1 (gal-1) is a 14-kDa laminin-binding galectin involved in several biologic events including regulation of cancer cell proliferation and adhesion to the matrix. In this study, we examined gal-1 expression in 30 human epithelial ovary carcinoma samples by Western and Northern blotting and by immunohistochemistry. Gal-1 mRNA levels were increased in more than 95% of the examined ovary carcinoma samples, compared with a wedge resection of a normal ovary. Immunohistochemical analysis of the samples demonstrated gal-1 expression in cancer epithelial cells from 17 of 30 samples, with a cytoplasmic pattern. Gal 1 immunostaining was significantly increased in the stroma associated with carcinoma cells compared with the normal, noninvaded stroma (p = 0.003). This pattern of expression was confirmed by examination of 12 other frozen epithelial ovary carcinomas, using in situ hybridization. Immunohistochemical staining of the specimens demonstrated colocalization of gal-1, laminin-1, and fibronectin. In vitro experiments were conducted to elucidate the potential biologic role of gal-1 in ovarian cancer progression. Gal-1 protein expression and release was detected in AZ364, SK-OV-3, and AZ224, but not in OVCAR-3, AZ419, and AZ382, human ovary carcinoma cell lines. Incubation of 84BR fibroblasts with conditioned media harvested from the ovary carcinoma cell lines induced an increased expression of gal-1 in the cultured fibroblasts in all cases except AZ419 and SK OV-3. High concentrations of gal-1 (100 micro g/ml) induced significantly decreased cell proliferation in all cell lines, as defined by bromodeoxyuridine incorporation. Additionally, recombinant gal-1 induced a dose-dependent increase in in vitro adhesion of AZ224, SK-OV-3, and AZ382 cells to laminin-1; adhesion to fibronectin was increased by gal-1 in OVCAR-3, AZ224, and SK-OV-3. No effect was observed in the other cases. Our data contribute to define a role for gal-1 during the interactions between human ovary carcinoma cells and host fibroblasts. PMID- 12649339 TI - Characterization of a novel breast carcinoma xenograft and cell line derived from a BRCA1 germ-line mutation carrier. AB - A human tumor xenograft (L56Br-X1) was established from a breast cancer axillary lymph node metastasis of a 53-year-old woman with a BRCA1 germ-line nonsense mutation (1806C>T; Q563X), and a cell line (L56Br-C1) was subsequently derived from the xenograft. The xenograft carries only the mutant BRCA1 allele and expresses mutant BRCA1 mRNA but no BRCA1 protein as determined by immunoprecipitation or Western blotting. The primary tumor, lymph node metastasis, and xenograft were hypodiploid by DNA flow cytometry, whereas the cell line displayed an aneuploidy apparently developed via polyploidization. Cytogenetic analysis, spectral karyotyping, and comparative genomic hybridization of the cell line revealed a highly complex karyotype with numerous unbalanced translocations. The xenograft and cell line had retained a somatic TP53 missense mutation (S215I) originating from the primary tumors, as well as a lack of immunohistochemically detectable expression of steroid hormone receptors, epidermal growth factor receptor, human epidermal growth factor receptor 2 (HER 2), and keratin 8. Global gene expression analysis by cDNA microarrays supported a correlation between the expression profiles of the primary tumor, lymph node metastasis, xenograft, and cell line. We conclude that L56Br-X1 and L56Br-C1 are useful model systems for studies of the pathogenesis and new therapeutic modalities of BRCA1-induced human breast cancer. PMID- 12649340 TI - Abnormal expression of the novel epidermal enzyme, glucosylceramide deacylase, and the accumulation of its enzymatic reaction product, glucosylsphingosine, in the skin of patients with atopic dermatitis. AB - To clarify mechanisms underlying acylceramide deficiency as an causative factor of the permeability barrier disruption seen in the skin of patients with atopic dermatitis (AD), we hypothesized and then demonstrated the presence of a novel epidermal enzyme, termed glucosylceramide (GC) deacylase. This enzyme hydrolyzes (acyl)GC at the N-acyl site to yield its lysoform, glucosylsphingosine (GS), instead of the formation of (acyl)ceramides by beta-glucocerebrosidase. Assays of enzymatic activity using [palmitic acid-(14)C] GC as a substrate revealed that extracts from the stratum corneum and from the epidermis (but not from the dermis) of patients with AD have the significantly higher potential to hydrolyze GC at the N-acyl site to release (14)C-labeled free fatty acid than of healthy controls. To determine the in vivo physiologic function of this novel enzyme, we measured the metabolic product GS in the upper stratum corneum. In both the involved and the uninvolved stratum corneum from patients with AD, there were significant increases in the amounts of GS compared with healthy controls and there was a significant inverse correlation with the decreased content of ceramides or ceramide-1 (acylceramide). Thus, collectively these results strongly suggest the physiologic relevance of GC deacylase to the acylceramide deficiency seen in the stratum corneum of patients with AD. PMID- 12649341 TI - Energetic characteristics of the new transthyretin variant A25T may explain its atypical central nervous system pathology. AB - Transthyretin (TTR) is a tetrameric protein that must misfold to form amyloid fibrils. Misfolding includes rate-limiting tetramer dissociation, followed by fast tertiary structural changes that enable aggregation. Amyloidogenesis of wild type (WT) TTR causes a late-onset cardiac disease called senile systemic amyloidosis. The aggregation of one of > 80 TTR variants leads to familial amyloidosis encompassing a collection of disorders characterized by peripheral neuropathy and/or cardiomyopathy. Prominent central nervous system (CNS) impairment is rare in TTR amyloidosis. Herein, we identify a new A25T TTR variant in a Japanese patient who presented with CNS amyloidosis at age 42 and peripheral neuropathy at age 44. The A25T variant is the most destabilized and fastest dissociating TTR tetramer published to date, yet, surprising, disease onset is in the fifth decade. Quantification of A25T TTR in the serum of this heterozygote reveals low levels relative to WT, suggesting that protein concentration influences disease phenotype. Another recently characterized TTR CNS variant (D18G TTR) exhibits strictly analogous characteristics, suggesting that instability coupled with low serum concentrations is the signature of CNS pathology and protects against early-onset systemic amyloidosis. The low A25T serum concentration may be explained either by impaired secretion from the liver or by increased clearance, both scenarios consistent with A25T's low kinetic and thermodynamic stability. Liver transplantation is the only known treatment for familial amyloid polyneuropathy. This is a form of gene therapy that removes the variant protein from serum preventing systemic amyloidosis. Unfortunately, the choroid plexus would have to be resected to remove A25T from the CSF-the source of the CNS TTR amyloid. Herein we demonstrate that small-molecule tetramer stabilizers represent an attractive therapeutic strategy to inhibit A25T misfolding and CNS amyloidosis. Specifically, 2-[(3,5 dichlorophenyl)amino]benzoic acid is an excellent inhibitor of A25T TTR amyloidosis in vitro. PMID- 12649342 TI - Molecular diagnosis of oligodendroglioma in paraffin sections. AB - Distinction of oligodendrogliomas from other gliomas is clinically important, but the histologic diagnosis of oligodendroglioma has been a difficult and notoriously subjective task. Testing for loss of heterozygosity (LOH) on chromosomal arms 1p and 19q, the genetic signature of oligodendroglioma, has emerged as a useful, objective adjunct to the traditional histologic evaluation. However, LOH testing of glioma specimens has not yet been widely implemented, presumably because of a lack of a practical LOH assay. We describe a 1p and 19q LOH assay suitable for routine diagnostics. In contrast to traditional microsatellite-based LOH analysis, we show that detection of LOH is usually possible even without normal tissue or blood from the same patient. A small area of tumor on a single paraffin section is sufficient for the assay. The assay protocol consists of a one-step DNA extraction, multiplex PCR for microsatellites on 1p and 19q, and capillary electrophoresis of the PCR products. LOH is detected by analysis of the allelic patterns and by integration of data from multiple highly polymorphic microsatellites. In a validation study on 19 gliomas, the results were concordant with results obtained by established methods and correlated well with histologic diagnoses. Because only a paraffin section is required, the pathologist can perform both the traditional histopathologic evaluation and this supporting molecular assay from the material at hand. PMID- 12649343 TI - Overexpression of human Dickkopf-1, an antagonist of wingless/WNT signaling, in human hepatoblastomas and Wilms' tumors. AB - Hepatoblastomas (HBs) represent the most frequent malignant liver tumors of childhood; yet little is known about the molecular pathogenesis and the alterations in expression patterns of these tumors. We used a suppression subtractive hybridization approach to identify new candidate genes that may play a role in HB tumorigenesis. cDNA species derived from corresponding liver and fetal liver were subtracted from HB cDNAs, and a series of interesting candidates were isolated that were differentially expressed. One of the transcripts overexpressed in HB was derived from the human Dickkopf-1 (hDkk-1) gene, which encodes a secreted protein acting as a potent inhibitor of the wingless/WNT signaling pathway. We examined the hDkk-1 expression levels in 32 HB biopsy specimens and in the corresponding liver samples, in 4 HB cell lines, and in a panel of other tumors and normal tissues using a differential PCR approach and Northern blotting. Eighty-one percent of the HBs but none of the normal pediatric or fetal liver tissues showed hDkk-1 expression. hDkk-1 transcripts were also present in 5 of 6 Wilms' tumors but only weakly detectable in 2 of 20 hepatocellular carcinoma samples and in 1 of 5 medulloblastoma cell lines; transcripts were absent in malignant gliomas and breast cancer. The central effector molecule in the WNT developmental control pathway is the beta-catenin protein. Interestingly, activating mutations of the beta-catenin gene have previously been identified in 48% of HBs, and more than 85% of HBs show accumulation of beta-catenin protein as the indicator for an activated pathway. The overexpression of the inhibitor Dkk-1 may therefore be related to uncontrolled wingless/WNT signaling and may represent a negative feedback mechanism. hDkk-1 expression represents a novel marker for HBs and Wilms' tumors. PMID- 12649344 TI - Epidermal growth factor-induced epithelio-mesenchymal transition in human breast carcinoma cells. AB - PMC42-LA cells display an epithelial phenotype: the cells congregate into pavement epithelial sheets in which E-cadherin and beta-catenin are localized at cell-cell borders. They abundantly express cytokeratins, although 5% to 10% of the cells also express the mesenchymal marker vimentin. Stimulation of PMC42-LA cells with epidermal growth factor (EGF) leads to epithelio-mesenchymal transition-like changes including up-regulation of vimentin and down-regulation of E-cadherin. Vimentin expression is seen in virtually all cells, and this increase is abrogated by treatment of cells with an EGF receptor antagonist. The expression of the mesenchyme-associated extracellular matrix molecules fibronectin and chondroitin sulfate proteoglycan also increase in the presence of EGF. PMC42-LA cells adhere rapidly to collagen I, collagen IV, and laminin-1 substrates and markedly more slowly to fibronectin and vitronectin. EGF increases the speed of cell adhesion to most of these extracellular matrix molecules without altering the order of adhesive preference. EGF also caused a time dependent increase in the motility of PMC42-LA cells, commensurate with the degree of vimentin staining. The increase in motility was at least partly chemokinetic, because it was evident both with and without chemoattractive stimuli. Although E-cadherin staining at cell-cell junctions disappeared in response to EGF, beta-catenin persisted at the cell periphery. Further analysis revealed that N-cadherin was present at the cell-cell junctions of untreated cells and that expression was increased after EGF treatment. N- and E-cadherin are not usually coexpressed in human carcinoma cell lines but can be coexpressed in embryonic tissues, and this may signify an epithelial cell population prone to epithelio-mesenchymal-like responses. PMID- 12649345 TI - Impaired gap junction connexin43 in Sertoli cells of patients with secretory azoospermia: a marker of undifferentiated Sertoli cells. AB - Gap junctions are intercellular channels formed of connexins (Cx) at appositional plasma membranes between adjacent cells that have been involved in the control of cell proliferation and differentiation. Altered Cx expression is implicated consistently in several human diseases and in tumorigenesis. Although Cx43 plays a critical role in Sertoli cell control of spermatogenesis, there is no evidence of its altered expression in human testicular pathologies. We show here that Cx43 mRNA expression was significantly reduced in testes of infertile patients with secretory azoospermia (p < 0.05) compared with testes displaying normal spermatogenesis (excretory azoospermic patients). In Sertoli cell-only syndrome, in situ hybridization and immunohistochemistry analyses indicated that Cx43 mRNA and protein were undetectable in Sertoli cells but were still present in the interstitial compartment. In a rat model of Sertoli cell-only syndrome, the lack of Cx43 in Sertoli cells was associated with an impairment of gap junction intercellular communication between adjacent Sertoli cells. These results reveal that Cx43 mRNA and protein expression are markedly impaired in Sertoli cells of infertile patients. This defect could be a new functional marker of undifferentiated Sertoli cells and could be related to the increased risk of testicular cancer recently described in the population of infertile men. PMID- 12649346 TI - Nucleus accumbens dopamine and the regulation of effort in food-seeking behavior: implications for studies of natural motivation, psychiatry, and drug abuse. AB - For several decades, it has been suggested that dopamine (DA), especially in nucleus accumbens, mediates the primary reinforcing characteristics of natural stimuli such as food, as well as drugs of abuse. Yet, several fundamental aspects of primary food reinforcement, motivation, and appetite are left intact after interference with accumbens DA transmission. Recent studies have shown that accumbens DA is involved in responsiveness to conditioned stimuli and activational aspects of motivation. In concurrent choice tasks, accumbens DA depletions cause animals to reallocate their choice behavior in the direction of instrumental behaviors that involve less effort. Also, an emerging body of evidence has demonstrated that the effects of accumbens DA depletions on instrumental food-seeking behavior can vary greatly depending upon the task. For example, some schedules of reinforcement are insensitive to the effects of DA depletions, whereas others are highly sensitive (e.g., large fixed ratios). Accumbens DA depletions slow the rate of operant responding, blunt the rate facilitating effects of moderate-sized ratios, and enhance the rate-suppressing effects of very large ratios (i.e., produce ratio strain). Accumbens DA may be important for enabling rats to overcome behavioral constraints, such as work related response costs, and may be critical for the behavioral organization and conditioning processes that enable animals to engage in vigorous responses, such as barrier climbing, or to emit large numbers of responses in ratio schedules in the absence of primary reinforcement. The involvement of accumbens DA in activational aspects of motivation has implications for energy-related disorders in psychiatry, as well as aspects of drug-seeking behavior. PMID- 12649347 TI - Metabolic control of excessive extracellular nucleotide accumulation by CD39/ecto nucleotidase-1: implications for ischemic vascular diseases. AB - Platelets are responsible for maintaining vascular integrity. In thrombocytopenic states, vascular permeability and fragility increase, presumably due to the absence of this platelet function. Chemical or physical injury to a blood vessel induces platelet activation and platelet recruitment. This is beneficial for the arrest of bleeding (hemostasis), but when an atherosclerotic plaque is ulcerated or fissured, it becomes an agonist for vascular occlusion (thrombosis). Experiments in the late 1980s cumulatively indicated that endothelial cell CD39 an ecto-ADPase-reduced platelet reactivity to most agonists, even in the absence of prostacyclin or nitric oxide. As discussed herein, CD39 rapidly and preferentially metabolizes ATP and ADP released from activated platelets to AMP, thereby drastically reducing or even abolishing platelet aggregation and recruitment. Since ADP is the final common agonist for platelet recruitment and thrombus formation, this finding highlights the significance of CD39. A recombinant, soluble form of human CD39, solCD39, has enzymatic and biological properties identical to the full-length form of the molecule and strongly inhibits human platelet aggregation induced by ADP, collagen, arachidonate, or TRAP (thrombin receptor agonist peptide). In sympathetic nerve endings isolated from guinea pig hearts, where neuronal ATP enhances norepinephrine exocytosis, solCD39 markedly attenuated norepinephrine release. This suggests that NTPDase (nucleoside triphosphate diphosphohydrolase) could exert a cardioprotective action by reducing ATP-mediated norepinephrine release, thereby offering a novel therapeutic approach to myocardial ischemia and its consequences. In a murine model of stroke, driven by excessive platelet recruitment, solCD39 reduced the sequelae of stroke, without an increase in intracerebral hemorrhage. CD39 null mice, generated by deletion of apyrase-conserved regions 2 to 4, exhibited a decrease in postischemic perfusion and an increase in cerebral infarct volume when compared with controls. "Reconstitution" of CD39 null mice with solCD39 reversed these changes. We hypothesize that solCD39 has potential as a novel therapeutic agent for thrombotic diatheses. PMID- 12649348 TI - A novel chymase inhibitor, 4-[1-([bis-(4-methyl-phenyl)-methyl]-carbamoyl)3-(2 ethoxy-benzyl)-4-oxo-azetidine-2-yloxy]-benzoic acid (BCEAB), suppressed cardiac fibrosis in cardiomyopathic hamsters. AB - Previously, we reported that levels of chymase activity and its mRNA in cardiac tissues were significantly increased along with progression of cardiac fibrosis in cardiomyopathic hamsters, but the involvement of chymase in the progression of fibrosis has been unclear. In cultured human fibroblasts, the concentration of transforming growth factor-beta in the supernatant of medium was significantly increased after injection of human chymase. Furthermore, human chymase dose dependently increased cell proliferation, and this chymase-dependent proliferation was completely suppressed by a chymase inhibitor, Suc-Val-Pro Phe(p)(OPh)(2) (10 micro M) or an anti-transforming growth factor-beta antibody (100 micro g/ml). In this study, we used Bio14.6 and F1B hamsters as cardiomyopathic and control hamsters, respectively. Cardiomyopathic hamsters were orally administered a novel chymase inhibitor, 4-[1-([bis-(4-methylphenyl) methyl]-carbamoyl)-3-(2-ethoxy-benzyl)-4-oxo-azetidine-2-yloxy]-benzoic acid (BCEAB; 100 mg/kg per day), or placebo from 5- to 45-week-old. In the placebo treated group, the cardiac chymase activity in cardiomyopathic hamsters 45 weeks old was significantly increased compared with that in control hamsters. BCEAB significantly reduced the cardiac chymase activity. The indexes (+dP/dt and dP/dt) of cardiac function were significantly improved by treatment with BCEAB. The mRNA levels of collagen I and collagen III in the placebo-treated hamsters were significantly reduced to 69.6 and 76.5% by treatment with BCEAB, respectively. The fibrotic area in cardiac tissues in the BCEAB-treated hamsters was significantly suppressed to 50.7% compared with that in the placebo-treated treated hamsters. Therefore, the activation of transforming growth factor-beta by chymase may play an important role in the progression of cardiac fibrosis and cardiac dysfunction in cardiomyopathy. PMID- 12649349 TI - Low-molecular-weight fucoidan promotes therapeutic revascularization in a rat model of critical hindlimb ischemia. AB - The therapeutic potential of low-molecular-weight (LMW) fucoidan, a sulfated polysaccharide extracted from brown seaweed devoid of direct antithrombin effect, was investigated in vitro and in a model of critical hindlimb ischemia in rat. In vitro results showed that LMW fucoidan enhanced fibroblast growth factor (FGF)-2 induced [(3)H]thymidine incorporation in cultured rat smooth muscle cells. Intravenous injection in rats of LMW fucoidan significantly increased the stromal derived factor (SDF)-1 level from 1.2 +/- 0.1 to 6.5 +/- 0.35 ng/ml in plasma. The therapeutic effect of LMW fucoidan (5 mg/kg/day), FGF-2 (1 micro g/kg/day), and LMW fucoidan combined with FGF-2 was assessed 14 days after induction of ischemia by 1) clinical evaluation of claudication, 2) tissue blood flow analysis, 3) histoenzymology of muscle metabolic activity, and 4) quantification of capillary density. Both LMW fucoidan and FGF-2 similarly improved residual muscle blood flow (62.5 +/- 6.5 and 64.5 +/- 4.5%, respectively) compared with the control group (42 +/- 3.5%, p < 0.0001). The combination of FGF-2 and LMW fucoidan showed further significant improvement in tissue blood flow (90.5 +/- 3%, p < 0.0001). These results were confirmed by phosphorylase activity, showing muscle regeneration in rats treated with the combination of FGF-2 and LMW fucoidan. Capillary density count increased from 9.6 +/- 0.7 capillaries/muscle section in untreated ischemic controls to 14.3 +/- 0.9 with LMW fucoidan, 14.5 +/ 0.9 with FGF-2, and 19.1 +/- 0.9 in combination (p < 0.001). Thus, LMW fucoidan potentiates FGF-2 activity, mobilizes SDF-1, and facilitates angiogenesis in a rat model. This natural compound could be of interest as an alternative for conventional treatment in critical ischemia. PMID- 12649350 TI - Neurokinin-1 receptor antagonists CP-96,345 and L-733,060 protect mice from cytokine-mediated liver injury. AB - Previously, we have shown that primary afferent sensory neurons are necessary for disease activity in T cell-mediated immune hepatitis in mice. In the present study, we analyzed the possible role of substance P (SP), an important proinflammatory neuropeptide of these nerve fibers, in an in vivo mouse model of liver inflammation. Liver injury was induced by bacterial lipopolysaccharide (LPS) in D-galactosamine (GalN)-sensitized mice. Depletion of primary afferent nerve fibers by neonatal capsaicin treatment down-regulated circulating levels of the proinflammatory cytokines tumor necrosis factor-alpha (TNFalpha) and interferon-gamma (IFNgamma) and protected mice from GalN/LPS-induced liver injury. Likewise, pretreatment of mice with antagonists of the SP-specific neurokinin-1 receptor (NK-1R), i.e., (2S,3S)-cis-2-(diphenylmethyl)-N-((2 methoxyphenyl)-methyl)-1-azabicyclo(2.2.2.)-octan-3-amine (CP-96,345) and (2S,3S)3-([3,5-bis(trifluoromethyl)phenyl]methoxy)-2-phenylpiperidine (L 733,060), dose dependently protected mice from GalN/LPS-induced liver injury. The presence of the NK-1R in the murine liver was demonstrated by reverse transcription-polymerase chain reaction, sequence analysis, and immunocytochemistry. NK-1R blockade reduced inflammatory liver damage, i.e., edema formation, neutrophil infiltration, hepatocyte apoptosis, and necrosis. To get further insight into the mechanism by which receptor blockade attenuated GalN/LPS-induced liver damage, we analyzed plasma levels and intrahepatic expression of TNFalpha, IFNgamma, interleukin (IL)-6, and IL-10. NK-1R blockade clearly inhibited GalN/LPS-induced production of TNFalpha and IFNgamma, whereas synthesis of the hepatoprotective cytokines IL-6 and IL-10 was increased. NK-1 receptor antagonists might be potent drugs for treatment of inflammatory liver disease, most likely by inhibiting SP effects. PMID- 12649351 TI - Delivery of peptide drugs to the brain by adenovirus-mediated heterologous expression of human oligopeptide transporter at the blood-brain barrier. AB - The feasibility of using adenovirus-mediated human oligopeptide transporter (hPEPT1) gene transfer to achieve peptide drug delivery to the brain across the blood-brain barrier was tested by examining the accumulation of model peptides in a rat brain endothelial cell line (RBEC1) and rat brain after transduction with a recombinant adenovirus encoding hPEPT1-enhanced yellow fluorescent protein fusion gene (AdhPEPT1-EYFP). In vitro uptake of [(3)H]GlySar was determined in RBEC1 transduced with AdhPEPT1-EYFP. In vivo, the accumulation of cefadroxil in rat brain was evaluated after transduction of AdhPEPT1-EYFP. At pH 6.0, the uptake of [(3)H]GlySar by RBEC1 transduced with AdhPEPT1-EYFP was increased 4-fold compared with that of nontransduced cells. At pH 7.4, uptake of [(3)H]GlySar in AdhPEPT1 EYFP transduced RBEC1 was 1.5 times higher than that of nontransduced cells. Unlabeled glycylsarcosine (10 mM) reduced the uptake of [(3)H]GlySar to a level comparable with that of nontransduced cells. At 30 min after intravenous administration of cefadroxil to rats transduced with AdhPEPT1-EYFP at 3.2 x 10(9) plaque-forming units/rat by an in situ brain perfusion method, the brain-to plasma concentration ratio (Kp) of cefadroxil was increased about 2 times compared with that of nontransduced or AdGFP (control vector)-transduced rats, although this was not statistically significant. In contrast, Kp of [(14)C]inulin, a marker for extracellular fluid space, remained unchanged after adenoviral transduction. In conclusion, our results suggest that adenovirus mediated heterologous expression of hPEPT1 in vivo could be a useful approach to deliver oligopeptides to the brain. PMID- 12649352 TI - The novel neuroprotective action of sulfasalazine through blockade of NMDA receptors. AB - Sulfasalazine is widely used to treat inflammatory diseases. Besides anti inflammatory actions such as blockade of nuclear factor-kappaB and cyclooxygenases, we found that 30 to 1000 micro M sulfasalazine dose dependently blocked N-methyl-D-aspartate receptor-mediated excitotoxicity without intervening kainate or alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid neurotoxicity. The neuroprotective effects of sulfasalazine were attributable to prevention of Ca(2+) influx and accumulation through N-methyl-D-aspartate receptors as a low-affinity antagonist. The systemic administration of sulfasalazine reduced neuronal death following transient cerebral and retinal ischemia in adult rat. The present findings suggest that the neuroprotective action of sulfasalazine can be therapeutically applied to halt devastating neuronal death following hypoxic ischemia, trauma, and neurodegenerative diseases. PMID- 12649353 TI - Pharmacological characterization of a novel nonpeptide antagonist radioligand, (+/-)-N-[2-methyl-4-methoxyphenyl]-1-(1-(methoxymethyl) propyl)-6-methyl-1H-1,2,3 triazolo[4,5-c]pyridin-4-amine ([3H]SN003) for corticotropin-releasing factor1 receptors. AB - The in vitro pharmacological profile of a novel small molecule corticotropin releasing factor 1 (CRF(1)) receptor antagonist, (+/-)-N-[2-methyl-4 methoxyphenyl]-1-(1-(methoxymethyl)propyl)-6-methyl-1H-1,2,3-triazolo[4,5 c]pyridin-4-amine (SN003), and the characteristics of its radioligand ([(3)H]SN003) are described. SN003 has high affinity and selectivity for CRF(1) receptors expressed in rat cortex, pituitary, and recombinant HEK293EBNA (HEK293e) cells with respective radiolabeled ovine CRF ([(125)I]oCRF) binding K(i) values of 2.5, 7.9, and 6.8 nM. SN003 was shown to be a CRF(1) receptor antagonist inasmuch as it inhibited CRF-induced cAMP accumulation in human CRF(1)HEK293e cells and CRF-stimulated adrenocorticotropin hormone release from rat pituitary cells without agonist activities. Significant decreases in the B(max) of [(125)I]oCRF binding by SN003 suggest that this antagonist is not simply competitive. To further explore the interaction of SN003 with the CRF(1) receptors, [(3)H]SN003 binding to rat cortex and human CRF(1)HEK293e cell membranes was characterized and shown to be reversible and saturable, with K(D) values of 4.8 and 4.6 nM, and B(max) values of 0.142 and 7.42 pmol/mg protein, respectively. The association and dissociation rate constants of [(3)H]SN003 (k(+1) 0.292 nM(-1) min(-1) and k(-1) 0.992 x 10(-2) min(-1)) were also assessed using human CRF(1)HEK293e cell membranes, giving an equilibrium dissociation constant of 3.4 nM. Moreover, [(3)H]SN003 binding displayed a single affinity state and insensitivity to 5'-guanylylimidodiphosphate, consistent with characteristics of antagonist binding. Incomplete inhibition of [(3)H]SN003 binding by CRF peptides also suggests that SN003 is not simply competitive with CRF at CRF(1) receptors. The distribution of [(3)H]SN003 binding sites was consistent with the expression pattern of CRF(1) receptors in rat brain regions. Small molecule CRF(1) antagonist radioligands like [(3)H]SN003 should enable a better understanding of small molecule interactions with the CRF(1) receptor. PMID- 12649354 TI - Amelioration of experimental autoimmune encephalomyelitis in Lewis rats by FTY720 treatment. AB - Experimental autoimmune encephalomyelitis (EAE) is a T-cell-dependent autoimmune disease that reproduces the inflammatory demyelinating pathology of multiple sclerosis (MS). We investigated the efficacy and mechanism of immunosuppression against EAE by administering 2-amino-[2-(4-octylphenyl) ethyl]-1,3-propanediol hydrochloride (FTY720) in Lewis rats immunized with myelin basic protein together with complete Freund's adjuvant. FTY720 treatment almost completely protected the rats against disease. The protection by FTY720 was associated with a dramatic reduction in the number of lymphocytes staining for T-cell receptors in the spinal cord as examined by immunohistochemistry. The mRNA expression of Th1 cytokines interleukin (IL)-2, IL-6, and interferon-gamma in the spinal cord was also reduced dramatically as assessed by reverse-transcription polymerase chain reaction. Furthermore, lymphocytes isolated from the spleen of FTY720-treated rats were transferred into naive recipient rats against EAE manifestation by reducing both disease incidence and clinical score. These results suggested that the protective anti-inflammatory effect of treatment with FTY720 was, to a large extent, due to the inhibition of encephalitogenic T-cell responses and/or their migration into the central nervous system and may be a potential candidate for use in treating patients with MS. PMID- 12649355 TI - The 3-hydroxy-3-methylglutaryl-CoA reductase inhibitor pravastatin reduces disease activity and inflammation in dextran-sulfate induced colitis. AB - The dextran sulfate (DSS) model of colitis causes intestinal injury sharing many characteristics with inflammatory bowel disease, e.g., leukocyte infiltration, loss of gut epithelial barrier, and cachexia. These symptoms are partly mediated by entrapped leukocytes binding to multiple endothelial adhesion molecules (MAdCAM-1, VCAM-1, ICAM-1, and E-selectin). Pravastatin, an 3-hydroxy-3 methylglutaryl (HMG)-CoA reductase inhibitor, has anti-inflammatory potency in certain inflammation models; therefore, in this study, we measured the effects of pravastatin in DSS-induced colitis. The administration of pravastatin (1 mg/kg) relieved DSS-induced cachexia, hematochezia, and intestinal epithelial permeability, with no effect on serum cholesterol. Histopathologically, pravastatin prevented leukocyte infiltration and gut injury. Pravastatin also blocked the mucosal expression of MAdCAM-1. DSS treatment promoted mucosal endothelial nitric-oxide synthase (eNOS) mRNA degradation, an effect that was blocked by pravastatin. Importantly, the protective effects of pravastatin in DSS induced colitis were not found in eNOS-deficient mice. Our results demonstrate that HMG-CoA reductase inhibitors preserve intestinal integrity in colitis, most likely via increased eNOS expression and activity, independent of cholesterol metabolism. PMID- 12649356 TI - Receptor occupancy of nonpeptide corticotropin-releasing factor 1 antagonist DMP696: correlation with drug exposure and anxiolytic efficacy. AB - 4-(1,3-Dimethoxyprop-2-ylamine)-2,7-dimethyl-8-(2,4-dichlorophenyl)-pyrazolo[1,5 a]-1,3,5-triazine (DMP696) is a highly selective and potent, nonpeptide corticotropin-releasing factor 1 (CRF(1)) antagonist. In this study, we measured in vivo CRF(1) receptor occupancy of DMP696 by using ex vivo ligand binding and quantitative autoradiography and explored the relationship of receptor occupancy with plasma and brain exposure and behavioral efficacy. In vitro affinity (IC(50)) of DMP696 to brain CRF(1) receptors measured using the brain section binding autoradiography in this study is similar to that assessed using homogenized cell membrane assays previously. The ex vivo binding assay was validated by demonstrating that potential underestimation of receptor occupancy with this procedure could be minimized by identifying an appropriate in vitro incubation time (40 min) based upon the dissociation kinetics of DMP696. Orally administrated DMP696 dose dependently occupied CRF(1) receptors in the brain, with ~60% occupancy at 3 mg/kg. In the defensive withdrawal test of anxiety, this dose of DMP696 produced approximately 50% reduction in the exit latency. The time course of plasma and brain drug levels paralleled that of receptor occupancy, with peak exposure at 90 min after dosing. The plasma-free concentration of DMP696 corresponding to 50% CRF(1) receptor occupancy (in vivo IC(50), 1.22 nM) was similar to the in vitro IC(50) (~1.0 nM). Brain concentrations of DMP696 were over 150-fold higher than the plasma-free levels. In conclusion, doses of DMP696 occupying over 50% brain CRF(1) receptors are consistent with doses producing anxiolytic efficacy in the defense withdrawal test of anxiety, and the IC(50) value estimated in vivo based on plasma-free drug concentrations is consistent with the in vitro IC(50) value. PMID- 12649357 TI - Beneficial effects of sampatrilat, a novel vasopeptidase inhibitor, on cardiac remodeling and function of rats with chronic heart failure following left coronary artery ligation. AB - Sampatrilat is a novel vasopeptidase inhibitor that may offer a greater benefit than traditional angiotensin-converting enzyme (ACE) inhibitors in the treatment of chronic heart failure (CHF). The present study was undertaken to determine whether sampatrilat improves hemodynamic function and cardiac remodeling through a direct action on the failing heart in rats with CHF following left coronary artery ligation (CAL). Sampatrilat (30 mg/kg a day) was administered orally to the animals from the 1st to 6th week after the operation. Sampatrilat reduced the mortality of the rats with CAL (20 versus 57% for untreated rats). Treatment with sampatrilat for 5 weeks suppressed tissue ACE and neutral endopeptidase (NEP) activities. Sampatrilat did not affect the arterial blood pressure, whereas it attenuated the CAL-induced increases in the left ventricular end-diastolic pressure, heart weight, and collagen content of the viable left ventricle. To assess the direct effects of sampatrilat on collagen synthesis, we measured the incorporation of [(3)H]proline into cultured cardiac fibroblasts. Sampatrilat at concentrations that inhibited NEP activity in vitro augmented the atrial natriuretic peptide-induced decrease in [(3)H]proline incorporation by the cells. In addition, sampatrilat prevented the angiotensin I-induced increase in [(3)H]proline incorporation, whereas captopril did not. The results suggest that long-term treatment with sampatrilat regresses cardiac remodeling in rats with CAL, which is associated with improvement of hemodynamic function. The mechanism by which sampatrilat improved cardiac remodeling may be attributable to the direct inhibition of cardiac fibrosis, possibly acting through the cardiac natriuretic peptide system. PMID- 12649358 TI - Increased relaxant action of forskolin and isoproterenol against muscarinic agonist-induced contractions in smooth muscle from M2 receptor knockout mice. AB - The ability of forskolin and isoproterenol to inhibit the contractile action of the muscarinic agonist, oxotremorine-M, was investigated in smooth muscle from wild-type and M(2) muscarinic receptor knockout mice. Forskolin (5.0 micro M) caused a significant reduction in the contractile activity of oxotremorine-M in ileum, trachea, and urinary bladder from both wild-type and M(2) muscarinic receptor knockout mice. This reduction in contractile activity was characterized by decreases in potency or maximal response, but not always both. Similar results were obtained with isoproterenol (1.0 micro M). The relaxant effects of forskolin in ileum, trachea, and urinary bladder from M(2) receptor knockout mice were approximately 3- to 9-fold greater than those observed in the same tissues from wild-type mice. Similar results were obtained with isoproterenol in ileum and urinary bladder, although the differences between wild-type and M(2) receptor knockout tissues were less than those observed with forskolin. In contrast, there was no significant difference between the relaxant effect of isoproterenol in trachea from wild-type and M(2) receptor knockout mice. In contrast to the results observed with oxotremorine-M as the contractile agent, forskolin and isoproterenol did not exhibit greater relaxant activity against KCl-induced contractions in M(2) receptor knockout mice compared with wild-type mice. These results suggest that a component of the contractile response to muscarinic agonists in smooth muscle involves an M(2) muscarinic receptor-mediated inhibition of the relaxant effects of agents that increase cAMP levels. PMID- 12649359 TI - Effect of chronic treatment with vitamin E on endothelial dysfunction in a type I in vivo diabetes mellitus model and in vitro. AB - Diabetes mellitus often leads to generalized vasculopathy. Because of the pathophysiological role of free radicals we investigated the effects of vitamin E. Twenty-eight rats were rendered diabetic by streptozotocin injection and were fed either with a diet with low (10 mg/kg of chow), medium (75 mg/kg of chow) or high amounts of vitamin E (1300 mg/kg of chow). Nine age-matched nondiabetic rats receiving 75 mg of vitamin E/kg chow served as controls. After 7 months, mesenteric microcirculation was investigated. Smooth muscle contractile function was not altered in diabetic versus nondiabetic vessels. Endothelial function was significantly reduced in diabetics; relaxation upon 1 micro M acetylcholine was reduced by 50% in diabetics with a medium and high vitamin E diet. In vitamin E deprived rats, a complete loss of endothelium-dependent relaxation was observed, and instead, acetylcholine elicited vasoconstriction. L-N(G)-Nitro-arginine induced vasoconstriction was reduced in small arteries in diabetics, which was not prevented by vitamin E, but was aggravated by vitamin E deprivation. In a subchronic endothelial cell culture model, cells were cultivated with 5 or 20 mM D-glucose for an entire cell culture passage (4 days) with or without vitamin E (20 mg/l versus 0.01 mg/l). Hyperglycemia led to significant reduction in basal and ATP-stimulated nitric oxide (NO)-production. Hyperglycemia-induced reduction in basal NO-release was significantly prevented by vitamin E, whereas reduction in stimulated NO-release was not influenced. NADPH-diaphorase activity was reduced by 40% by hyperglycemia, which was completely prevented by vitamin E. We conclude that 1) vitamin E has a potential to prevent partially hyperglycemia induced endothelial dysfunction, 2) under in vivo conditions vitamin E deficiency enhanced diabetic endothelial dysfunction dramatically, and 3) positive effects of vitamin E may be attenuated with a longer disease duration. PMID- 12649360 TI - Renal function in a rat model of analgesic nephropathy: effect of chloroquine. AB - The antimalaria drug chloroquine is often taken against a background of analgesic nephropathy caused by nonsteroidal anti-inflammatory drugs such as paracetamol (acetaminophen). Chloroquine has marked effects on the normal kidney and stimulates an increase in plasma vasopressin via nitric oxide. The aim of this study was to determine the renal action of chloroquine in a model of analgesic nephropathy. Sprague-Dawley rats (n = 6-8/group) were treated with paracetamol (500 mg kg(-1) day(-1)) for 30 days in drinking water to induce analgesic nephropathy; control rats received normal tap water. Under intraval anesthesia (100 mg kg(-1)) rats were infused with 2.5% dextrose for 3 h to equilibrate and after a control hour they received either vehicle, chloroquine (0.04 mg h(-1)), N(omega)-nitro-L-arginine methyl ester (L-NAME, nitric-oxide synthase inhibitor, 60 micro g kg(-1) h(-1)) or combined chloroquine and L-NAME over the next hour. Plasma was collected from a parallel group of animals for vasopressin radioimmunoassay. Long-term paracetamol treatment resulted in a decrease in glomerular filtration rate (p < 0.05), sodium excretion (p < 0.001), and urine osmolality (p < 0.001), but no change in urine flow rate compared with untreated animals. Chloroquine administration in paracetamol treated rats induced a significant reduction (p < 0.05) in urine flow rate and a significant increase in plasma vasopressin (p < 0.001). These effects were blocked by coadministration of L-NAME and thus seem to be mediated by a pathway involving nitric oxide. However, these responses contrast with the chloroquine-induced diuresis previously observed in untreated rats, possibly reflecting paracetamol inhibition of renal prostaglandin synthesis and consequent moderation of vasopressin's action. PMID- 12649361 TI - L-homocysteine sulfinic acid and other acidic homocysteine derivatives are potent and selective metabotropic glutamate receptor agonists. AB - Moderate hyperhomocysteinemia is associated with several diseases, including coronary artery disease, stroke, Alzheimer's disease, schizophrenia, and spina bifida. However, the mechanisms for their pathogenesis are unknown but could involve the interaction of homocysteine or its metabolites with molecular targets such as neurotransmitter receptors, channels, or transporters. We discovered that L-homocysteine sulfinic acid (L-HCSA), L-homocysteic acid, L-cysteine sulfinic acid, and L-cysteic acid are potent and effective agonists at several rat metabotropic glutamate receptors (mGluRs). These acidic homocysteine derivatives 1) stimulated phosphoinositide hydrolysis in the cells stably expressing the mGluR1, mGluR5, or mGluR8 (plus Galpha(qi9)) and 2) inhibited the forskolin induced cAMP accumulation in the cells stably expressing mGluR2, mGluR4, or mGluR6, with different potencies and efficacies depending on receptor subtypes. Of the four compounds, L-HCSA is the most potent agonist at mGluR1, mGluR2, mGluR4, mGluR5, mGluR6, and mGluR8. The effects of the four agonists were selective for mGluRs because activity was not discovered when L-HCSA and several other homocysteine derivatives were screened against a large panel of cloned neurotransmitter receptors, channels, and transporters. These findings imply that mGluRs are candidate G-protein-coupled receptors for mediating the intracellular signaling events induced by acidic homocysteine derivatives. The relevance of these findings for the role of mGluRs in the pathogenesis of homocysteine mediated phenomena is discussed. PMID- 12649362 TI - Mixed cocaine agonist/antagonist properties of (+)-methyl 4beta-(4-chlorophenyl) 1-methylpiperidine-3alpha-carboxylate, a piperidine-based analog of cocaine. AB - The present study investigated the pharmacological properties of a piperidine based novel cocaine analog, namely, (+)-methyl 4beta-(4-chlorophenyl)-1 methylpiperidine-3alpha-carboxylic acid [(+)-CPCA]. Like cocaine, (+)-CPCA inhibited rat synaptosomal dopamine and norepinephrine uptake with high affinity, but was 33-fold less potent than cocaine in inhibiting serotonin uptake. Like cocaine, (+)-CPCA is a locomotor stimulant, although it was less potent and efficacious than cocaine. Importantly, pretreatment with (+)-CPCA dose dependently blocked the locomotor stimulant effects of cocaine in rats. (+)-CPCA completely substituted for cocaine in drug discrimination tests, although it was about 3 times less potent than cocaine. It was also self-administered by rats. Unexpectedly, (+)-CPCA did not enhance cocaine-induced convulsions in mice. As expected from rodent studies, rhesus monkeys readily self-administered (+)-CPCA. However, compared with cocaine, (+)-CPCA showed limited reinforcing properties in rats as assessed by both fixed and progressive ratio intravenous drug self administration tests. These results collectively suggest that (+)-CPCA has an atypical pharmacological profile having both cocaine-like "agonist" and some cocaine "antagonist" properties. These properties of (+)-CPCA suggest that it may have utility in the treatment of cocaine craving and dependence. PMID- 12649363 TI - A kinetic evaluation of the absorption, efflux, and metabolism of verapamil in the autoperfused rat jejunum. AB - P-glycoprotein (P-gp)-mediated drug efflux from the apical membrane of enterocytes is believed to modulate intestinal cytochrome P450 3A (CYP3A) metabolism by altering substrate access to the CYP3A enzyme. This interplay between P-gp and CYP3A was investigated in a rat in situ model of intestinal permeation, where a recirculating luminal perfusion of the jejunum was coupled with mesenteric vein blood collection to simultaneously monitor the uptake, transport, and metabolism of the P-gp and CYP3A substrate, verapamil. Transport of intact verapamil into the venous blood was increased by 160, 84, and 160%, and the intestinal extraction ratio on passage across the jejunum was reduced by 15, 24, and 97% by inhibitors of P-gp [PSC833 ([3'-keto-Me-Bmt(1)]-[Val(2)] cyclosporin)], CYP3A (midazolam), or P-gp and CYP3A (ketoconazole), respectively, when present in the luminal perfusate and compared with control experiments. Compartmental kinetic analysis of the data revealed that inhibition of P-gp did not affect the rate constant describing verapamil metabolism but, rather, increased the intestinal uptake of verapamil and stimulated a disproportionate increase in verapamil transport into the venous blood. The increase in verapamil transport, in the absence of changes to metabolism, reduced the intestinal extraction ratio. This finding may be explained by saturation of intracellular verapamil binding sites within the intestinal tissue in response to increased verapamil uptake resulting from P-gp inhibition. The current findings confirm previous in vitro and theoretical approaches which suggest that P-gp can modulate the extent of intestinal extraction of P-gp/CYP3A substrates. PMID- 12649364 TI - Hypothalamic-pituitary-thyroid axis and sympathetic nervous system involvement in hyperthermia induced by 3,4-methylenedioxymethamphetamine (Ecstasy). AB - An acute and potentially life-threatening complication associated with the recreational use of the 3,4-methylenedioxymethamphetamine (MDMA, Ecstasy) is hyperthermia. In the present study, Sprague-Dawley rats treated with MDMA (40 mg/kg s.c.) responded with a significant increase (maximal at 1 h) in rectal and skeletal muscle temperatures that lasted for at least 3 h post-treatment. Hypophysectomized (HYPO) and thyroparathyroidectomized (TX) animals treated with MDMA (40 mg/kg s.c.) did not become hyperthermic and in fact displayed a significant hypothermia. The HYPO and TX animals were also resistant to the serotonergic neurotoxic effects of MDMA assessed by serotonin measurements 4 to 7 days later in the striatum and hippocampus. MDMA (40 mg/kg s.c.) induced a significant increase in thyroxine levels 1 h post-treatment. Thyroid hormone replacement in TX animals returned the hyperthermic response seen after MDMA. Prazosin, an alpha(1)-antagonist (0.2 mg/kg i.p.), administered 30 min before MDMA significantly attenuated the MDMA-induced increase in rectal temperature, but had no effect on skeletal muscle temperature. Cyanopindolol, a beta(3) antagonist (4 mg/kg s.c.), administered 30 min before MDMA (40 mg/kg s.c.) significantly attenuated the increase in skeletal muscle temperature, but had no effect on the rise in rectal temperature. The combination of prazosin and cyanopindolol resulted in an abolishment of MDMA-induced hyperthermia. The mechanisms of thermogenesis induced by MDMA seem to result from an interaction between the hypothalamic-pituitary-thyroid axis and the sympathetic nervous system, wherein mechanisms leading to core and skeletal muscle hyperthermia after MDMA exposure seem to be differentially regulated by alpha(1)- and beta(3) adrenergic receptors. PMID- 12649365 TI - Is 11beta-hydroxysteroid dehydrogenase type 1 a therapeutic target? Effects of carbenoxolone in lean and obese Zucker rats. AB - In liver and adipose tissue, 11beta-hydroxysteroid dehydrogenase type 1 (11beta HSD1) regenerates glucocorticoids from inactive 11-keto metabolites. Pharmacological inhibition or transgenic disruption of 11beta-HSD1 attenuates glucocorticoid action and increases insulin sensitivity. Increased adipose 11beta HSD1 may also contribute to the metabolic complications of obesity. Here, we examine the effects of inhibition of 11beta-HSDs with carbenoxolone in obese insulin-resistant Zucker rats, a strain in which tissue-specific dysregulation of 11beta-HSD1 (increased in adipose, decreased in liver) mirrors changes in human obesity. Six-week-old male rats were treated orally with carbenoxolone (50 mg/kg/day) or water (1 ml/kg/day) for 3 weeks. Carbenoxolone inhibited 11beta HSD1 activity in liver (25 +/- 3 versus 52 +/- 2% conversion in lean; 18 +/- 3 versus 35 +/- 3% in obese; p < 0.01) but not in adipose tissue or skeletal muscle. Carbenoxolone had no effect on weight gain or food intake, did not affect plasma glucose during an oral glucose tolerance test, and increased the plasma insulin response to glucose. However, high-density lipoprotein cholesterol was increased by carbenoxolone in obese animals (1.52 +/- 0.24 versus 1.21 +/- 0.26 mM; p < 0.03). Carbenoxolone did not inhibit hepatic inactivation of glucocorticoid by 5beta-reductase and had no significant effect on plasma corticosterone levels. In conclusion, carbenoxolone provides a model for liver specific inhibition of 11beta-HSD1, which results in improved lipid profile, in Zucker obese rats. Failure to inhibit 11beta-HSD1 in adipose tissue and/or skeletal muscle may explain the lack of effect on glucose tolerance and obesity. Inhibition of adipose 11beta-HSD1 is probably necessary to gain the maximum benefit of an 11beta-HSD1 inhibitor. PMID- 12649366 TI - Activation of kappa-opioid receptors inhibits pruritus evoked by subcutaneous or intrathecal administration of morphine in monkeys. AB - Pruritus (itch sensation) is the most common side effect associated with spinal administration of morphine given to humans for analgesia. A variety of agents have been proposed as antipruritics with poorly understood mechanisms and they are effective with variable success. kappa-Opioid agonists possess several actions that are opposite to micro -opioid agonists. We proposed to investigate the role of kappa-opioid receptors (KORs) in morphine-induced scratching and antinociception in monkeys. Scratching responses were counted by observers blinded to treatment. Antinociception was measured by a warm water (50 degrees C) tail-withdrawal assay. Pretreatment with low doses of trans-(+/-)-3,4-dichloro-N methyl-N-(2-[1-pyrrolidinyl]-cyclohexyl)-benzeneacetamide (U-50488H) (0.032-0.18 mg/kg s.c.), a selective KOR agonist, dose dependently suppressed the s.c. morphine dose-effect curve for scratching and potentiated s.c. morphine-induced antinociception. In addition, s.c. U-50488H attenuated i.t. morphine (10 and 32 micro g)-induced scratching while maintaining or enhancing i.t. morphine-induced antinociception. The combination of s.c. or i.t. morphine with low doses of U 50488H did not cause sedation. More importantly, pretreatment with 3.2 mg/kg nor binaltorphimine, a selective KOR antagonist, blocked the effects of s.c. U-50488H on both s.c. and i.t. morphine-induced scratching. These results indicate that activation of KOR attenuates morphine-induced scratching without interfering with antinociception in monkeys. This mechanism-based finding provides functional evidence in support of the clinical potential of KOR agonists as antipruritics in the presence of MOR agonist-induced pruritus. PMID- 12649367 TI - Individual differences in cocaine-induced locomotor sensitization in low and high cocaine locomotor-responding rats are associated with differential inhibition of dopamine clearance in nucleus accumbens. AB - Behavioral sensitization to cocaine reflects neuroadaptive changes that intensify drug effects. However, repeated cocaine administration does not induce behavioral sensitization in all male Sprague-Dawley rats. Because cocaine inhibits the dopamine (DA) transporter (DAT), we investigated whether altered DAT function contributes to these individual differences. Freely moving rats had electrochemical microelectrode/microcannulae assemblies chronically implanted in the nucleus accumbens so that exogenous DA clearance signals were recorded simultaneous with behavior. The peak DA signal amplitude (A(max)) and efficiency of clearance (k) were used as indices of in vivo DAT function. Low and high cocaine responders (LCRs and HCRs, respectively) were identified based on their locomotor responsiveness to an initial injection of cocaine (10 mg/kg i.p.). Consistent with DAT inhibition, cocaine elevated A(max) and reduced k in HCRs, but not in LCRs. The same dose of cocaine was administered for six additional days and after a 7-day withdrawal. Baseline behavioral and dopamine clearance indices were unaltered by repeated cocaine or after withdrawal. Only LCRs expressed cocaine-induced sensitized locomotor activation, and this was accompanied by cocaine-induced elevations in A(max) and reductions in k. These sensitized responses to cocaine persisted in LCRs after withdrawal. In contrast, neither locomotor nor electrochemical responses were altered by repeated saline administration or a saline challenge after repeated cocaine administration, suggesting that conditioning did not significantly contribute. Our results suggest that increased DAT inhibition by cocaine is associated with locomotor sensitization and that DAT serves as a common substrate for mediating both the initial and sensitized locomotor responsiveness to cocaine. PMID- 12649368 TI - Differential regulation of GABA B receptor subunit expression and function. AB - The GABA(B) receptor is a G protein-coupled heterodimer composed of GABA(B1) and GABA(B2) subunits. In the present study, experiments were undertaken to examine the relationship between GABA(B) receptor function and subunit expression in the rat lumbar spinal cord following pharmacological and physiological manipulation of this receptor system. Although formalin-induced hind paw inflammation increases the production of GABA(B1) and GABA(B2) protein in the spinal cord within 24 h, there is no change in receptor function, as measured by the baclofen stimulated guanosine 5'-O-(3-[(35)S]thiotriphosphate) ([(35)S]GTPgammaS) binding assay. Conversely, although chronic (7 days) administration of baclofen, a GABA(B) receptor agonist, abolishes baclofen-stimulated [(35)S]GTPgammaS binding in the spinal cord tissue, causes tolerance to the sedative and antinociceptive effects of the drug, increases the number of formalin-induced hind paw flinches, and induces mechanical hyperalgesia, this treatment had no effect on the levels of GABA(B1) or GABA(B2) mRNAs in the lumbar spinal cord. The results indicate a lack of concordance between expression of GABA(B1) and GABA(B2) subunits and GABA(B) receptor function, suggesting these subunit proteins may serve multiple functions in the cells. Moreover, these findings indicate that nongenomic mechanisms are primarily responsible for the GABA(B) receptor desensitization that occurs during prolonged exposure to receptor agonist. PMID- 12649369 TI - Inhibition of P-glycoprotein by newer antidepressants. AB - Pharmacokinetic drug-drug interactions often occur at the level of P-glycoprotein (Pgp). To study possible interactions caused by the newer antidepressants we investigated citalopram, fluoxetine, fluvoxamine, paroxetine, reboxetine, sertraline, and venlafaxine and their major metabolites desmethylcitalopram, norfluoxetine, paroxetine-metabolite (paroxetine-M), desmethylsertraline, N desmethylvenlafaxine, and O-desmethylvenlafaxine for their ability to inhibit Pgp. Pgp inhibition was studied by a fluorometric assay using calcein acetoxymethylester as Pgp substrate and two different cell systems: L-MDR1 cells (model for human Pgp) and primary porcine brain capillary endothelial cells (pBCECs, model for the blood-brain barrier). Both cell systems proved to be suitable for the evaluation of Pgp inhibitory potency of drugs. All antidepressants tested except O-desmethylvenlafaxine showed Pgp inhibitory activity with sertraline, desmethylsertraline, and paroxetine being the most potent, comparable with the well known Pgp inhibitor quinidine. In L-MDR1 cells fluoxetine, norfluoxetine, fluvoxamine, reboxetine, and paroxetine-M revealed intermediate Pgp inhibition and citalopram, desmethylcitalopram, venlafaxine, and N-desmethylvenlafaxine were only weak inhibitors. The ranking order was similar in pBCECs. The fact that some of the compounds tested exert Pgp inhibitor effects at similar concentrations as quinidine suggests that pharmacokinetic drug-drug interactions between the newer antidepressants and Pgp substrates should now be thoroughly studied in vivo. PMID- 12649370 TI - Structure-activity study of LVV-hemorphin-7: angiotensin AT4 receptor ligand and inhibitor of insulin-regulated aminopeptidase. AB - The decapeptide LVV-hemorphin-7 binds with high affinity to the angiotensin IV (Ang IV) receptor (AT(4) receptor), eliciting a number of physiological effects, including cellular proliferation and memory enhancement. We have recently shown that the AT(4) receptor is identical to insulin-regulated aminopeptidase (IRAP) and that both LVV-hemorphin-7 and Ang IV inhibit the catalytic activity of IRAP. In the current study, a series of alanine-substituted and N- or C-terminally modified analogs of LVV-hemorphin-7 were evaluated for their abilities to compete for (125)I-Ang IV binding in sheep adrenal and cerebellar membranes. Selected analogs were also analyzed for binding to recombinant human IRAP and inhibition of IRAP aminopeptidase activity. C-Terminal deletions of LVV-hemorphin-7 resulted in modest changes in affinity for IRAP, whereas deletion of the first three N terminal residues abolished binding. Monosubstitutions of Tyr(4) and Trp(6) with alanine resulted in a 10-fold reduction in affinity. Competition binding studies using recombinant human IRAP demonstrated the same rank order of affinity as obtained for the ovine tissues. All LVV-hemorphin-7 analogs tested, except for Leu-Val-Val-Tyr, inhibit the cleavage of the synthetic substrate, leucine beta naphthylamide, by IRAP, with K(i) values between 56 and 620 nM. We find that the Val(3) residue is crucial for LVV-hemorphin-7 binding to IRAP, whereas the C terminal domain seems to play a minor role. The current study highlights the minimal residues necessary for binding and inhibition of IRAP and provides a basis to design peptidomimetic analogs for experimental and potentially clinical use. PMID- 12649371 TI - Dextromethorphan protects dopaminergic neurons against inflammation-mediated degeneration through inhibition of microglial activation. AB - Inflammation in the brain has increasingly been recognized to play an important role in the pathogenesis of several neurodegenerative disorders, including Parkinson's disease and Alzheimer's disease. Inflammation-mediated neurodegeneration involves activation of the brain's resident immune cells, the microglia, which produce proinflammatory and neurotoxic factors, including cytokines, reactive oxygen intermediates, nitric oxide, and eicosanoids that impact on neurons to induce neurodegeneration. Hence, identification of compounds that prevent microglial activation may be highly desirable in the search for therapeutic agents for inflammation-mediated neurodegenerative diseases. In this study, we report that dextromethorphan (DM), an ingredient widely used in antitussive remedies, reduced the inflammation-mediated degeneration of dopaminergic neurons through inhibition of microglial activation. Pretreatment (30 min) of rat mesencephalic neuron-glia cultures with DM (1-10 micro M) reduced, in a dose-dependent manner, the microglia-mediated degeneration of dopaminergic neurons induced by lipopolysaccharide (LPS, 10 ng/ml). Significant neuroprotection by DM was also evident when DM was applied to cultures up to 60 min after the addition of LPS. The neuroprotective effect of DM was attributed to inhibition of LPS-stimulated microglial activation because DM significantly inhibited the LPS-induced production of tumor necrosis factor-alpha, nitric oxide, and superoxide free radicals. This conclusion was further supported by the finding that DM failed to prevent 1-methyl-4-phenylpyridinium- or beta-amyloid peptide (1-42)-induced dopaminergic neurotoxicity in neuron-enriched cultures. In addition, because LPS did not produce any significant increase in the release of excitatory amino acids from neuron-glia cultures and N-methyl-D-aspartate antagonist dizocilpine maleate failed to afford significant neuroprotection, it is unlikely that the neuroprotective effect of DM is mediated through N-methyl-D aspartate receptors. These results suggest that DM may be a promising therapeutic agent for the treatment of Parkinson's disease. PMID- 12649372 TI - Delta-aminolevulinic acid transport in cancer cells of the human extrahepatic biliary duct. AB - This study was performed to characterize the transport of the endogenous photosensitizer delta-aminolevulinic acid in tumor cells of the extrahepatic biliary duct. Uptake of [(3)H]delta-aminolevulinic acid into human cholangiocarcinoma SK-ChA-1 cells was linear for up to 10 min, independent of a Na(+) gradient, but stimulated 3- to 4-fold by an inwardly directed H(+) gradient. Uptake of delta-aminolevulinic acid was mediated by a single transport system with an apparent affinity (K(t)) of 2.1 mM and a maximal velocity (V(max)) of 60.1 nmol. 10 min(-1). mg of protein(-1). Glycylsarcosine, alanylalanine, and cefadroxil strongly inhibited the [(3)H]delta-aminolevulinic acid uptake with K(i) values of 1.3, 0.2, and 3.6 mM, respectively. In contrast, gamma aminobutyric acid, glycine, L-glutamic acid, and L-aspartic acid (all 10 mM) had no effect on the total [(3)H]delta-aminolevulinic acid uptake, neither at pH 6.0 nor at pH 7.5. Applying a Dixon type of experiment and the ABC test revealed that glycylsarcosine and delta-aminolevulinic acid are transported via the same system, PEPT1. Treatment of the cells with phorbol 12-myristate 13-acetate, a phorbol ester that activates protein kinase C, resulted in a significant inhibition of the transport rate. This inhibition could be blocked by cotreatment with staurosporine. We conclude that delta-aminolevulinic acid is transported by the H(+)/peptide cotransporter PEPT1 into epithelial cells of the extrahepatic biliary duct. delta-Aminolevulinic acid can be accumulated specifically in bile duct tumor cells before photodynamic therapy. PMID- 12649373 TI - N-[3-(R)-tetrahydrofuranyl]-6-aminopurine riboside, an A1 adenosine receptor agonist, antagonizes catecholamine-induced lipolysis without cardiovascular effects in awake rats. AB - Elevated serum nonesterified free fatty acid (NEFA) concentrations are detrimental to both the mechanical and electrical function of the heart. A(1) adenosine receptor agonists are potent and efficacious inhibitors of lipolysis; however, their cardiovascular effects have limited their use to lower serum NEFA. Our objective was to determine whether the antilipolytic effect of N-[3-(R) tetrahydrofuranyl]-6-aminopurine riboside (CVT-510), an A(1) agonist, could be distinguished from its bradycardia effect and demonstrated in rats with normal or elevated serum NEFA. Rats were instrumented with telemetry transmitters for continuous recording of heart rate, and catheterized, for delivery of drugs and blood sampling. CVT-510 caused a rapid and sustained dose-dependent decrease in NEFA at doses that did not cause bradycardia (2, 5, and 20 micro g/kg). Significant bradycardia was observed at 50 micro g/kg. Norepinephrine (NE) increased NEFA from 0.5 +/- 0.01 to 0.9 +/- 0.2 mM and this effect lasted for 2 h. CVT-510 (10 micro g/kg) given at 40 min postinjection of NE reversed the rise in NEFA (69% reduction). When CVT-510 (20 micro g/kg) was given 15 min before a 30-min long infusion of NE, the lipolytic response to NE was prevented. To mimic the antilipolytic effect of CVT-510 in awake rats, hearts were perfused with palmitate at concentrations similar to those observed in the in vivo studies (0.8 and 0.2 mM), which decreased myocardial oxygen consumption (MVO(2)) by 11%. Thus, CVT-510 at doses > or =5-fold lower than those that slow heart rate caused a marked and sustained lowering of normal or elevated NEFA, that when mimicked in vitro decreased MVO(2) and would be expected to improve cardiac efficiency. PMID- 12649374 TI - Significant role of ceramide pathway in experimental gastric ulcer formation in rats. AB - Ceramides have emerged as key participants in the signaling pathway of cytokines and apoptosis. We previously revealed that phorbol 12-myristate 13-acetate (PMA) induced experimental ulcers in rat gastric mucosa. In this study, we investigated the role of ceramide in ulcer formation and its relation to the activation of transcription factors and apoptosis. PMA was subserosally injected to rat glandular stomach. Fumonisin B1 (FB1), an inhibitor of ceramide synthase, was administered together with the PMA. The time course of ceramide content was quantified using thin layer chromatography and the number of apoptotic cells was determined by immunohistochemistry. The activation of transcription factor nuclear factor-kappaB (NF-kappaB) or activator protein-1 (AP-1) was evaluated using an electrophoretic mobility shift assay. The administration of FB1 attenuated PMA-induced gastric ulcer formation in a dose-dependent manner. Before the ulcers became obvious, the ceramide content (C18 and C24 ceramide) increased significantly in the gastric wall. The activation of NF-kappaB and AP-1 and an increase in the number of apoptotic cells were also observed. Both of these were significantly inhibited by the coadministration of FB1. However, NF-kappaB inhibitors attenuated gastric ulcer formation without affecting the ceramide content or the number of apoptotic cells. Ceramide formation in the stomach significantly contributes to PMA-induced tissue damage, possibly via the activation of transcription factors and an increase in apoptosis in the gastric mucosa. However, after the increase in ceramide levels, the NF-kappaB and apoptosis pathways may be separately involved in ulcer formation. PMID- 12649375 TI - Inhibition of cyclooxygenases reduces complement-induced glomerular epithelial cell injury and proteinuria in passive Heymann nephritis. AB - In the passive Heymann nephritis (PHN) model of rat membranous nephropathy, complement induces glomerular epithelial cell injury and proteinuria, which is partially mediated by eicosanoids. Glomerular cyclooxygenase (COX)-1 and -2 are up-regulated in PHN and contribute to prostanoid generation. In the current study, we address the role of COX isoforms in proteinuria, using the nonselective COX inhibitor indomethacin and the COX-2-selective inhibitor 5,5-dimethyl-3-(3 fluorophenyl)-4-(4-methylsulphonyl)phenyl-2(5H)-furanone (DFU). Four groups of rats with PHN were treated twice daily, from day 7 through 14 with vehicle, 1 mg/kg DFU, 10 mg/kg DFU, or 2 mg/kg indomethacin. Vehicle-treated rats with PHN showed significant proteinuria on day 14 (163 +/- 15 mg/d, n = 19), compared with normal rats (10 +/- 4 mg/d, n = 3, p < 0.001). Treatment with DFU (1 or 10 mg/kg) reduced proteinuria significantly (by ~33%), compared with vehicle, but to a lesser extent than indomethacin (56% reduction). Glomerular eicosanoid generation was reduced significantly in the DFU and indomethacin groups, compared with vehicle. There were no significant differences among vehicle- or DFU-treated groups in [(3)H]inulin clearance, or in glomerular expression of COX-1 and -2. DFU did not affect the autologous immune response. In cultured rat glomerular epithelial cells, COX inhibition reduced complement-induced cytotoxicity, and this reduction was reversed by the thromboxane A(2) analog 9,11-dideoxy 9alpha,11alpha-methanoepoxyprostaglandin F(2alpha) (U46619). Thus, in experimental membranous nephropathy, selective inhibition of COX-2 reduces proteinuria, without adversely affecting renal function. However, inhibition of both COX-1 and -2 is required to achieve a maximum cytoprotective and antiproteinuric effect. PMID- 12649376 TI - In vivo mechanistic studies on the metabolic activation of 2-phenylpropionic acid in rat. AB - Two alternative metabolic pathways, acyl glucuronidation and acyl-CoA formation, are implicated in the generation of reactive acylating metabolites of carboxylic acids. Here, we describe studies that determine the relative importance of these two pathways in the metabolic activation of a model substrate, 2-phenylpropionic acid (2-PPA), in vivo in rats. Male Sprague-Dawley rats were pretreated with and without (-)-borneol (320 mg/kg i.p.), an inhibitor of acyl glucuronidation, or trimethylacetic acid (TMA, 500 mg/kg i.p.), an inhibitor of acyl-CoA formation, before receiving 2-PPA (racemic, 130 mg/kg). After administration of 2-PPA, livers were collected over a 2-h period and analyzed for 2-PPA acyl glucuronidation and 2-PPA-CoA formation by high-performance liquid chromatography. Covalent binding was measured by scintillation counting of washed liver protein precipitates. Results showed that pretreatment with TMA led to a 49% decrease in covalent binding of 2-PPA to liver proteins, when a 64% decrease in the exposure of 2-PPA-CoA was observed. Conversely, 95% inhibition of acyl glucuronidation by (-)-borneol, led to a 23% decrease in covalent binding to protein. These results suggest that metabolic activation by 2-PPA-CoA formation contributes to covalent adduct formation to protein in vivo to a greater extent than metabolic activation by acyl glucuronidation for this model substrate. PMID- 12649377 TI - Amiodarone-induced postrepolarization refractoriness suppresses induction of ventricular fibrillation. AB - It is still incompletely understood why amiodarone is such a potent antiarrhythmic drug. We hypothesized that chronic amiodarone treatment produces postrepolarization refractoriness (PRR) without conduction slowing and that PRR modifies the induction of ventricular arrhythmias. In this study, the hearts of 15 amiodarone-pretreated (50 mg/kg p.o. for 6 weeks) rabbits and 13 controls were isolated and eight monophasic action potentials were simultaneously recorded from the epicardium and endocardium of both ventricles. Steady-state action potential duration (APD), conduction times, refractory periods, and dispersion of action potential durations were determined during programmed stimulation and during 50 Hz burst stimuli, and related to arrhythmia inducibility. Amiodarone prolonged APD by 12 to 15 ms at pacing cycle lengths of 300 to 600 ms (p < 0.05) but did not significantly increase conduction times or dispersion of APD. Amiodarone prolonged refractoriness more than action potential duration, resulting in PRR (refractory period - APD at 90% repolarization, 14 +/- 10 ms, p < 0.05 versus controls). PRR curtailed the initial sloped part of the APD restitution curve by 20%. During burst stimulation, pronounced amiodarone-induced PRR (40 +/- 15 ms, p < 0.05 versus controls) reduced the inducibility of ventricular arrhythmias (p < 0.05 versus controls). Furthermore, in 35% of bursts only monomorphic ventricular tachycardias and no longer ventricular fibrillation were inducible in amiodarone treated hearts (p < 0.05 versus controls). Chronic amiodarone treatment prevents ventricular tachycardias by inducing PRR without much conduction slowing, thereby curtailing the initial part of APD restitution. PRR without conduction slowing is a desirable feature of drugs designed to prevent ventricular arrhythmias. PMID- 12649378 TI - Ethanol differentially enhances hippocampal GABA A receptor-mediated responses in protein kinase C gamma (PKC gamma) and PKC epsilon null mice. AB - Ethanol intoxication results partly from actions of ethanol at specific ligand gated ion channels. One such channel is the GABA(A) receptor complex, although ethanol's effects on GABA(A) receptors are variable. For example, we found that hippocampal neurons from selectively bred mice and rats with high hypnotic sensitivity to ethanol have increased GABA(A) receptor-mediated synaptic responses during acute ethanol treatment compared with mice and rats that display low behavioral sensitivity to ethanol. Here we investigate whether specific protein kinase C (PKC) isozymes modulate hypnotic and GABA(A) receptor sensitivity to ethanol. We examined acute effects of ethanol on GABA(A) receptor mediated inhibitory postsynaptic currents (IPSCs) in mice lacking either PKCgamma (PKCgamma(-/-)) or PKCepsilon (PKCepsilon(-/-)) isozymes and compared the results to those from corresponding wild-type littermates (PKCgamma(+/+) and PKCepsilon(+/+)). GABA(A) receptor-mediated IPSCs were evoked in CA1 pyramidal neurons by electrical stimulation in stratum pyramidale, and the responses were recorded in voltage-clamp mode using whole-cell patch recording techniques. Ethanol (80 mM) enhanced the IPSC response amplitude and area in PKCgamma(+/+) mice, but not in the PKCgamma(-/-) mice. In contrast, ethanol markedly potentiated IPSCs in the PKCepsilon(-/-) mice, but not in PKCepsilon(+/+) littermates. There was a positive correlation between ethanol potentiation of IPSCs and the ethanol-induced loss of righting reflex such that mice with larger ethanol-induced increases in GABA(A) receptor-mediated IPSCs also had higher hypnotic sensitivity to ethanol. These results suggest that PKCgamma and PKCepsilon signaling pathways reciprocally modulate both ethanol enhancement of GABA(A) receptor function and hypnotic sensitivity to ethanol. PMID- 12649379 TI - Ca v 1.3 is preferentially coupled to glucose-stimulated insulin secretion in the pancreatic beta-cell line INS-1. AB - L-Type Ca(2+) channel blockers inhibit glucose and KCl-stimulated insulin secretion by pancreatic beta cells. However, the role of the two distinct L-type channels expressed by beta cells, Ca(v)1.2 and Ca(v)1.3, in this process is not clear. Therefore, we stably transfected INS-1 cells with two mutant channel constructs, Ca(v)1.2DHPi or Ca(v)1.3 DHPi. Whole-cell patch-clamp recordings demonstrated that both mutant channels are insensitive to dihydropyridines (DHPs), but are blocked by diltiazem. INS-1 cells expressing Ca(v)1.3/DHPi maintained glucose- and KCl-stimulated insulin secretion in the presence of DHPs, whereas cells expressing Ca(v)1.2/DHPi demonstrated DHP resistance to only KCl induced secretion. INS-1 cells were also stably transfected with cDNAs encoding the intracellular loop between domains II and III of either Ca(v)1.2 or Ca(v)1.3 (Ca(v)1.2/II-III or Ca(v)1.3/II-III). Glucose- and KCl-stimulated insulin secretion in Ca(v)1.2/II-III cells were not different from untransfected INS-1 cells. However, glucose-stimulated insulin secretion was completely inhibited and KCl-stimulated secretion was substantially resistant to inhibition by DHPs, but sensitive to omega-agatoxin IVA in Ca(v)1.3/II-III cells. Moreover, the L-type channel agonist FPL 64176 markedly enhanced KCl-stimulated secretion by Ca(v)1.3/II-III cells. Together, our results suggest that Ca(2+) influx via Ca(v)1.3 is preferentially coupled to glucose-stimulated insulin secretion in INS 1 cells. PMID- 12649380 TI - Moment analysis of metabolic heterogeneity: conjugation of benzoate with glycine in rat liver studied by multiple indicator dilution technique. AB - Metabolic zonation was assessed with the multiple indicator dilution (MID) technique in the single-pass perfused rat liver with use of moment analysis of the formed metabolite (M) data. During single-pass, retrograde rat liver perfusion with 17 Awake M benzoate, a bolus containing tracer preformed metabolite (PM) [(3)H]hippurate was injected rapidly into the hepatic vein at 20 min postperfusion, followed by injection of a second bolus containing [(14)C]benzoate at 30 min. Both doses also contained noneliminated reference indicators ((51)Cr-labeled RBCs, (125)I-labeled albumin, [(14)C]- or [(3)H]sucrose, and (2)H(2)O). The steady-state extraction ratio of benzoate, the area under the curve (AUC) and its mean transit time (MTT) during retrograde flow were identical to those previously observed for prograde flow. Values of AUC(PM) and MTT(PM) and AUC(M) were also similar to previously published prograde data, but the MTT(M) with retrograde perfusion was smaller than that for prograde perfusion. This, according to theory based on the tubes-in-series model, was consistent with perivenous enrichment of glycination activity when transport of drug was even and when the ratio of drug influx/efflux coefficient exceeded that for metabolite. Similar benzoate transport in periportal, homogeneous and perivenous isolated rat hepatocytes existed, and the influx/efflux coefficients (partition ratio) of benzoate from MID indeed exceeded that of hippurate. However, metabolism by zonal hepatocytes failed to reveal the anticipated metabolic zonation, and this is likely due to the shallow gradient of metabolic activity. The study demonstrates that moment theory is useful in delineating the perivenous enrichment of glycine conjugation activity. PMID- 12649381 TI - Stress-induced suppression of the immune system after withdrawal from chronic cocaine. AB - Recent evidence suggests that withdrawal from cocaine shares similarities to the stress response. Here, we examine whether withdrawal from chronic cocaine produces immune system alterations and whether the hypothalamic-pituitary-adrenal axis is involved. Sprague-Dawley male rats received cocaine (10 mg/kg i.p., b.i.d.) or saline, followed by 2 h, 1, 2, 4, 6, and 14 days of withdrawal. Proliferation responses of peripheral blood lymphocytes to concanavalin A were significantly suppressed at the 2-h, 1- and 2-day time points, and persisted for up to 6 days during withdrawal from chronic cocaine. Flow cytometric analysis revealed no significant differences in the immunophenotype of blood lymphocytic populations of T cells, B cells, or monocytes at 2 or 6 days of withdrawal from cocaine. Consistent with the suppression in cellular immunity observed in the in vitro response, the in vivo delayed-type hypersensitivity response was also significantly decreased in cocaine withdrawing animals. Plasma corticosterone levels were significantly elevated 2 and 24 h after cessation of cocaine but returned to basal values by 2 days of withdrawal. The suppressive effects of cocaine withdrawal were no longer observed in either adrenalectomized animals or those treated with the glucocorticoid receptor antagonist mifepristone (RU486), when administered during the first 2 days of withdrawal. These data argue that repeated exposure to cocaine followed by withdrawal leads to an activation of the neuroendocrine stress response, which alters cellular immunity during the initial withdrawal phase and may contribute to an increased susceptibility to infection. PMID- 12649382 TI - Gabexate mesilate, a synthetic protease inhibitor, inhibits lipopolysaccharide induced tumor necrosis factor-alpha production by inhibiting activation of both nuclear factor-kappaB and activator protein-1 in human monocytes. AB - Gabexate mesilate, a synthetic protease inhibitor, was shown to be effective in treating patients with sepsis-associated disseminated intravascular coagulation in which tumor necrosis factor-alpha (TNF-alpha) plays a critical role. We demonstrated that gabexate mesilate reduced lipopolysaccharide (LPS)-induced tissue injury by inhibiting TNF-alpha production in rats. In the present study, we analyzed the mechanism(s) by which gabexate mesilate inhibits LPS-induced TNF alpha production in human monocytes in vitro. Gabexate mesilate inhibited the production of TNF-alpha in monocytes stimulated with LPS. Gabexate mesilate inhibited both the binding of nuclear factor-kappaB (NF-kappaB) to target sites and the degradation of inhibitory kappaBalpha. Gabexate mesilate also inhibited both the binding of activator protein-1 (AP-1) to target sites and the activation of mitogen-activated protein kinase pathways. These observations strongly suggest that gabexate mesilate inhibited LPS-induced TNF-alpha production in human monocytes by inhibiting activation of both NF-kappaB and AP-1. Inhibition of TNF alpha production by gabexate mesilate might explain at least partly its therapeutic effects in animals given LPS and those in patients with sepsis. PMID- 12649383 TI - In vivo modulation of intestinal CYP3A metabolism by P-glycoprotein: studies using the rat single-pass intestinal perfusion model. AB - P-Glycoprotein (P-gp) has been hypothesized to modulate intestinal drug metabolism by increasing the exposure of drug to intracellular CYP3A through repeated cycles of drug absorption and efflux. The rat single-pass intestinal perfusion model was used to study this interplay in vivo. N-Methyl piperazine-Phe homoPhe-vinylsulfone phenyl (K77), a peptidomimetic cysteine protease inhibitor (CYP3A/P-gp substrate), and midazolam (CYP3A substrate) were each perfused through a segment of rat ileum alone and with the P-gp inhibitor N-(4-[2-(1,2,3,4 tetrahydro-6,7-dimethoxy-2-isoquinolinyl)-ethyl]-phenyl)-9,10-dihydro-5-methoxy-9 oxo-4-acridine carboxamine (GG918). Samples were obtained continuously from the outlet perfusate and the mesenteric vein at 5-min intervals for 40 to 60 min. The parent drug and two main metabolites of K77 (N-desmethyl and N-oxide) and midazolam (1-OH and 4-OH) were quantitated by liquid chromatography/mass spectrometry. K77 appearance in the mesenteric blood (P(blood) = 5 +/- 3 x 10(-6) cm/s) was increased 3-fold with GG918, whereas midazolam permeability (P(blood) = 1.1 +/- 0.3 x 10(-4) cm/s) was unchanged by GG918. K77 metabolites were preferentially excreted into the lumen, 4-OH midazolam was found equally in lumen and blood, and 1-OH was mainly excreted into blood. The extent of metabolism was estimated by calculating the fraction metabolized = 1 - P(blood)/P(lumen) and the extraction ratio (ER) determined from the direct measurement of known metabolites as ER = sum metabolites(all)/(sum metabolites(all) + drug in blood). When P-gp was inhibited, the fraction metabolized for K77 was decreased (95 to 85%) and the ER tended toward a decrease, whereas no differences in either parameter were observed for midazolam (not a P-gp substrate). These data support a role for P-gp in modulating the extent of intestinal metabolism in vivo by controlling drug access to the enzyme. PMID- 12649384 TI - Contribution of individual cytochrome P450 isozymes to the O-demethylation of the psychotropic beta-carboline alkaloids harmaline and harmine. AB - The psychotropic beta-carboline alkaloids, showing high affinity for 5 hydroxytryptamine, dopamine, benzodiazepine, and imidazoline receptors and the stimulation of locus coeruleus neurons, are formed endogenously from tryptophan derived indolealkylamines through the Pictet-Spengler condensation with aldehydes in both plants and mammals. Cytochromes P450 1A1 (18.5), 1A2 (20), and 2D6 (100) catalyzed the O-demethylation of harmaline, and CYP1A1 (98.5), CYP1A2 (35), CYP2C9 (16), CYP2C19 (30), and CYP2D6 (115) catalyzed that of harmine (relative activities). The dehydrogenation/aromatization of harmaline to harmine was not carried out by aromatase (CYP19), CYP1A2, CYP2C9, CYP2D6, CYP3A4, pooled recombinant cytochromes P450, or human liver microsomes (HLMs). Kinetic parameters were calculated for the O-demethylations mediated by each isozyme and by pooled HLMs. K(cat) (min(-1)) and K(m) Awake M) values for harmaline were: CYP1A1, 10.8 and 11.8; CYP1A2, 12.3 and 13.3; CYP2C9, 5.3 and 175; CYP2C19, 10.3 and 160; and CYP2D6, 39.9 and 1.4. Values for harmine were: CYP1A1, 45.2 and 52.2; CYP1A2, 9.2 and 14.7; CYP2C9, 11.9 and 117; CYP2C19, 21.4 and 121; and CYP2D6, 29.7 and 7.4. Inhibition studies using monoclonal antibodies confirmed that CYP1A2 and CYP2D6 were the major isozymes contributing to both harmaline (20% and 50%, respectively) and harmine (20% and 30%) O-demethylations in pooled HLMs. The turnover numbers for CYP2D6 are among the highest ever reported for a CYP2D6 substrate. Finally, CYP2D6-transgenic mice were found to have increased harmaline and harmine O-demethylase activities as compared with wild-type mice. These findings suggest a role for polymorphic CYP2D6 in the pharmacology and toxicology of harmine and harmaline. PMID- 12649385 TI - Antidepressant-like effects of kappa-opioid receptor antagonists in the forced swim test in rats. AB - We showed previously that cAMP response element-binding protein (CREB) within the nucleus accumbens (NAc) of rats regulates immobility in the forced swim test (FST), an assay used to study depression. Because CREB regulates expression of dynorphin (which acts at kappa-opioid receptors) in NAc neurons, these findings raised the possibility that kappa-receptors mediate immobility behaviors in the FST. Here, we report that i.c.v. administration of the kappa-antagonist nor binaltorphimine dose dependently decreased immobility in the FST, suggesting that it has antidepressant-like effects. Implicating a specific effect at kappa receptors, similar antidepressant-like effects were seen after treatment with either of two novel, structurally dissimilar kappa-antagonists: 5' guanidinonaltrindole, which was effective after i.c.v. but not systemic treatment, and 5'-acetamidinoethylnaltrindole (ANTI), which was potent and effective after systemic treatment. The behavioral effects of the kappa antagonists resembled those of tricyclic antidepressants (desipramine) and selective serotonin reuptake inhibitors (fluoxetine and citalopram). Conversely, systemic administration of the kappa-agonist [5alpha,7alpha,8beta]-N-methyl-N-[7 [1-pyrrolidinyl]-1-oxaspiro[4.5]dec8-yl]-benzenacetamide (U-69593) dose dependently increased immobility in the FST, consistent with prodepressant-like effects. The effects of the kappa-ligands in the FST were not correlated with nonspecific effects on locomotor activity. Furthermore, the most potent and effective kappa-antagonist (ANTI) did not affect the rewarding impact of lateral hypothalamic brain stimulation at a dose with strong antidepressant-like effects. These findings are consistent with the hypothesis that CREB-mediated induction of dynorphin in the NAc "triggers" immobility behavior in the FST. Furthermore, they raise the possibility that kappa-antagonists may have efficacy as antidepressants, but lack stimulant or reward-related effects. PMID- 12649386 TI - Suppression of hepatic CYP3A1/2 and CYP2C11 by cyclosporine is not mediated by altering growth hormone levels. AB - Cyclosporine (CsA) suppresses drug metabolism by decreasing cytochrome P450 (P450) enzyme levels in rat liver. Growth hormone (GH) is known to pretranslationally regulate P450 expression. Thus, the suppression of P450 by CsA may involve GH as an intermediate. To address this question, we examined the effects of administering exogenous GH via twice daily subcutaneous injections and in conjunction with chronic subcutaneous CsA administration for 14 days on hepatic P450 expression. CsA alone decreased CYP3A1/2 and CYP2C11 significantly, in a manner similar to that previously found. When administered in the absence of CsA, GH also suppressed CYP3A1/2 and CYP2C11 protein levels as compared with GH vehicle. In the presence of CsA, GH did not cause further suppression of either CYP3A1/2 or CYP2C11 expression when compared with CsA treatment with GH vehicle. Testosterone in vitro catalytic assays confirmed that CsA and GH separately cause significant decreases in activity levels. Also, the concomitant administration of GH and CsA caused lowered production of 16alpha-, 2alpha-, 6beta-, and 2beta hydroxytestosterone as compared with the administration of GH with CsA vehicle and as compared with the administration of GH vehicle with CsA. This study shows that GH is a dominating factor over CsA in determining hepatic P450 expression and activity. In addition, CsA does not seem to alter GH levels as a mediating event in suppressing P450 expression and activity. Since CsA given in combination with GH further suppressed P450 activity as compared with CsA given in combination with vehicle, this suggests that changes in hormonal status are likely to be one of the many factors that is responsible for the lack of a clear association between cyclosporine dosing and markers of toxicity. PMID- 12649387 TI - Piribedil enhances frontocortical and hippocampal release of acetylcholine in freely moving rats by blockade of alpha 2A-adrenoceptors: a dialysis comparison to talipexole and quinelorane in the absence of acetylcholinesterase inhibitors. AB - In a dialysis procedure not requiring perfusate addition of acetylcholinesterase inhibitors to "boost" basal levels of acetylcholine (ACh), the influence of the antiparkinson agent piribedil upon levels of ACh in frontal cortex and dorsal hippocampus of freely moving rats was compared with those of other antiparkinson drugs and selective ligands at alpha(2)-adrenoceptors (ARs). Suggesting a tonic, inhibitory influence of alpha(2A)-ARs upon cholinergic transmission, the alpha(2) AR agonist 5-bromo-6-[2-imidazolin-2-yl-amino]-quinoxaline tartrate (UK14,304), and the preferential alpha(2A)-AR agonist guanabenz reduced levels of ACh. They were elevated by the antagonists 2(2-methoxy-1,4 benzodioxan-2-yl)-2-imidazoline HCl (RX821002) and atipamezole and by the preferential alpha(2A)-AR antagonist 2 (2H-(1-methyl-1,3-dihydroisoindole)methyl)-4,5-dihydroimidazole (BRL44008). In contrast, trans-2,3,9,13b-tetrahydro-1,2-dimethyl-1H-dibenz[c,f]imidazo[1,5 a]azepine (BRL41992) and prazosin, preferential alpha(2B/2C)-AR antagonists, were inactive. The dopaminergic agonist and antiparkinson agent piribedil, which behaves as an antagonist at alpha(2)-ARs, dose dependently increased extracellular levels of ACh. This action was absent upon pretreatment with a maximally effective dose of RX821002. On the other hand, a further dopaminergic agonist and antiparkinson agent, talipexole, which possesses agonist properties at alpha(2)-ARs, dose dependently reduced levels of ACh. This action was also blocked by RX821002. In contrast to piribedil and talipexole, quinelorane, which interacts with dopaminergic receptors but not alpha(2)-ARs, failed to affect ACh levels. Finally, in analogy to the frontal cortex, piribedil likewise elicited a dose-dependent increase in extracellular levels of ACh in the dorsal hippocampus. In conclusion, in distinction to talipexole and quinelorane, and reflecting its antagonist properties at alpha(2A)-ARs, piribedil reinforces cholinergic transmission in the frontal cortex and dorsal hippocampus of freely moving rats. These actions may be related to its facilitatory influence upon cognitive function. PMID- 12649388 TI - An orally bioavailable small molecule antagonist of CRTH2, ramatroban (BAY u3405), inhibits prostaglandin D2-induced eosinophil migration in vitro. AB - Ramatroban (Baynas, BAY u3405), a thromboxane A(2) (TxA(2)) antagonist marketed for allergic rhinitis, has been shown to partially attenuate prostaglandin (PG)D(2)-induced bronchial hyperresponsiveness in humans, as well as reduce antigen-induced early- and late-phase inflammatory responses in mice, guinea pigs, and rats. PGD(2) is known to induce eosinophilia following intranasal administration, and to induce eosinophil activation in vitro. In addition to the TxA(2) receptor, PGD(2) is known as a ligand for the PGD(2) receptor, and the newly identified G-protein-coupled chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2). To fully characterize PGD(2)-mediated inflammatory responses relevant to eosinophil activation, further analysis of the mechanism of action of ramatroban has now been performed. PGD(2)-stimulated human eosinophil migration was shown to be mediated exclusively through activation of CRTH2, and surprisingly, these effects were completely inhibited by ramatroban. This is also the first report detailing an orally bioavailable small molecule CRTH2 antagonist. Our findings suggest that clinical efficacy of ramatroban may be in part mediated through its action on this Th2-, eosinophil-, and basophil specific chemoattractant receptor. PMID- 12649389 TI - Insulin and glucagon regulation of glutathione S-transferase expression in primary cultured rat hepatocytes. AB - Diabetes is a major cause of morbidity and mortality, and complications resulting from diabetes have been attributed in part to increased oxidative stress. Glutathione S-transferases (GSTs) constitute a major protective mechanism against oxidative stress. Studies of the expression and activity of GSTs during diabetes are inconclusive, with both increased and decreased GST expression being reported in vivo. Insulin and glucagon effects on GST expression and the signaling pathway involved in the glucagon regulation of GST expression were examined in primary cultured rat hepatocytes. The addition of insulin resulted in the elevation of alpha-class GST protein levels, whereas alpha- and pi-class GST protein levels were markedly decreased in hepatocytes cultured with glucagon. In contrast, mu class GST protein expression was unaffected by insulin or glucagon treatment. Insulin concentrations >/=1 nM resulted in increased GST activities and alpha class GST protein levels, whereas glucagon concentrations >/=20 nM decreased alpha- and pi-class protein levels and activity. Treatment of cells with 8-bromo cAMP or dibutyryl-cAMP also resulted in decreased alpha- and pi-class GST protein levels. Pretreatment with N-[2-(4-bromocinnamylamino)ethyl]-5-isoquinoline sulfonamide (H89), a selective inhibitor of protein kinase A, before glucagon addition markedly attenuated the glucagon effect. This study demonstrates that insulin and glucagon regulate, in an opposing manner, the expression of alpha class GSTs and that glucagon completely inhibits pi-class GST expression in vitro, suggesting that hepatic GST expression may be decreased during diabetes. Furthermore, the present study implicates cAMP and protein kinase A in mediating the inhibitory effect of glucagon on GST expression. PMID- 12649390 TI - Sexual dimorphism in cafeteria diet-induced hypertension is associated with gender-related difference in renal leptin receptor down-regulation. AB - Plasma leptin levels are elevated in obesity suggesting a pathophysiologic role of this hormone in obesity and related disorders, such as hypertension. Furthermore, despite excess leptin levels, leptin satiety action is blunted in obesity suggesting the occurrence of central leptin resistance. As leptin acts on the kidney to induce natriuresis, renal leptin receptor alterations could lead to a defect in sodium excretion and hence to hypertension. Therefore, the present study investigated renal leptin receptor (Ob-Ra and Ob-Rb) mRNA and leptin binding capacities in diet-induced hypertension. Feeding male, female, and testosterone-treated female rats a cafeteria diet for 10 weeks increased body fat mass, plasma insulin, and leptin levels. Furthermore, although male and testosterone-treated female cafeteria-fed rats were hypertensive, the female rats fed the same diet failed to develop elevated blood pressure. In renal medulla, Ob Ra and Ob-Rb mRNA levels were unchanged after cafeteria diet feeding in all groups; however, binding analysis revealed Ob-R protein down-regulation exclusively in hypertensive rats. Moreover, renal Ob-R densities were inversely correlated to plasma leptin concentrations in male rats and testosterone-treated female rats but not in female rats. These findings demonstrate the existence of differences in renal Ob-R binding capacities, which are correlated to hypertension. PMID- 12649391 TI - Ambient temperature influences core body temperature response in rat lines bred for differences in sensitivity to 8-hydroxy-dipropylaminotetralin. AB - Agonist-induced decrease in core body temperature has commonly been used as a measure of serotonin1A (5-HT(1A)) receptor sensitivity in mood disorder. The thermoregulatory basis for 5-HT(1A) receptor agonist-induced temperature responses in humans and rats remains unclear. Therefore, the influence of ambient temperature on 5-HT(1A) receptor-mediated decreases in core body temperature were measured in rat lines bred for high (HDS) or low (LDS) sensitivity to the selective 5-HT(1A) receptor agonist 8-hydroxy-dipropylaminotetralin (8-OH-DPAT). HDS and LDS rats were injected with either saline, 0.25 or 0.50 mg/kg 8-OH-DPAT at ambient temperatures of 10.5, 24, 30, or 37.5 degrees C, and core temperature was measured by radiotelemetry. For both lines, the thermic response to acute 8 OH-DPAT was greatest at 10.5 degrees C and decreased in magnitude as ambient temperature increased to 30 degrees C, consistent with hypothermia. HDS rats displayed a greater hypothermic response than LDS rats at 10.5, 24, and 30 degrees C. At 37.5 degrees C, LDS rats showed a lethal elevation of temperature in response to 0.50 mg/kg 8-OH-DPAT. All thermic responses to 8-OH-DPAT, including the lethality, were effectively blocked by pretreatment with the 5 HT(1A) receptor antagonist WAY100635, suggesting line differences in thermoregulatory circuits that are influenced by 5-HT(1A) receptor activation. Following repeated injection of 8-OH-DPAT, the magnitude of the hypothermic response decreased in both lines at 10.5 degrees C, but increased in HDS rats treated with 0.50 mg/kg 8-OH-DPAT at 30 and 37.5 degrees C. This pattern was reversed in HDS rats following 8-OH-DPAT challenge at 24 degrees C, suggesting that a compensatory thermoregulatory response accounts for changes in the hypothermic response to chronic 8-OH-DPAT. PMID- 12649392 TI - Repeated exposures to subthreshold doses of chlorpyrifos in rats: hippocampal damage, impaired axonal transport, and deficits in spatial learning. AB - Organophosphorus (OP) compounds are detectable in the environment for years after use and endanger many populations. Although the effects of acutely toxic doses of many OP compounds are well described, much less is known about repeated low-level exposures. The purpose of these studies was to further evaluate potential toxicological effects of the extensively used OP pesticide chlorpyrifos (CPF) in rats. CPF, across a range of subthreshold doses (i.e., for acute toxicity), reduced rearing and sniffing activity and the magnitude of weight gain over 14 days of repeated exposure. Performance in a spatial learning task was impaired after 14 days of exposure to CPF (18.0 and 25.0 mg/kg) when testing was initiated 24 h after the last injection but not after a 14-day washout. However, inhibition of both fast anterograde and retrograde axonal transport was observed for up to 20 days after exposure to 25.0 mg/kg CPF. Studies using hippocampal cultures indicated that 8 days of continuous exposure to the parent compound, CPF (> or =100 micro M), resulted in cell toxicity and death. Furthermore, a dose (2.5 mg/kg) of CPF that had no effects on weight gain or memory performance when administered 5 days per week over 38 days impaired forelimb grip strength in the later days of testing. Collectively, these results indicate that repeated exposures to subthreshold doses of CPF may lead to growth retardation, behavioral abnormalities, and muscle weakness. Some of these symptoms may be attributed to effects of the OP on axonal transport. PMID- 12649393 TI - Human urocortin II, a selective agonist for the type 2 corticotropin-releasing factor receptor, decreases feeding and drinking in the rat. AB - Corticotropin-releasing factor (CRF) has been hypothesized to modulate consummatory behavior through the Type 2 CRF (CRF(2)) receptor. However, behavioral functions subserved by the CRF(2) receptor remain poorly understood. Recently, human urocortin II (hUcn II), a selective CRF(2) receptor agonist, was identified. To study the effects of this neuropeptide on ingestive behavior, we examined the effects of centrally infused hUcn II (i.c.v. 0, 0.01, 0.1, 1.0, 10.0 micro g) on the microstructure of nose-poke responding for food and water in nondeprived, male rats. Malaise-inducing properties of the peptide were monitored using conditioned taste aversion (CTA) testing. To identify potential sites of action, central induction of Fos protein expression was examined. hUcn II dose dependently reduced the quantity and duration of responding for food and water at doses lower (0.01-1.0 micro g) than that forming a CTA (10 micro g). Effects were most evident during hours 4 to 6 of the dark cycle. Meal pattern analysis showed that hUcn II potently (0.1 micro g) increased the satiating value of food. Rats ate and drank smaller and shorter meals without changing meal frequency. Rats also ate more slowly. hUcn II induced Fos in regions involved in visceral sensory processing and autonomic/neuroendocrine regulation and resembling those activated by appetite suppressants. hUcn II is a promising neuropeptide for investigating the role of the CRF(2) receptor in ingestive behavior. PMID- 12649394 TI - Role of cytochrome P4501B1 in benzo[a]pyrene bioactivation to DNA-binding metabolites in mouse vascular smooth muscle cells: evidence from 32P-postlabeling for formation of 3-hydroxybenzo[a]pyrene and benzo[a]pyrene-3,6-quinone as major proximate genotoxic intermediates. AB - Benzo[a]pyrene (BP), a polycylic aromatic hydrocarbon (PAH), is a potent atherogen and carcinogen in laboratory animals. Since genotoxic mechanisms may contribute to the development of atherosclerosis by PAHs, we have tested the hypotheses that: 1) BP induces DNA adducts in mouse aortic smooth muscle cells (SMCs); 2) 3-hydroxybenzo[a]pyrene (3-OH-BP) and benzo[a]pyrene-3,6-quinone (BPQ) are proximate genotoxic metabolites; and 3) cytochrome P4501B1 (CYP1B1) mediates the activation of BP and its metabolites to ultimate genotoxic intermediates. Cultured mouse aortic SMCs were treated with BP, 3-OH-BP, or BPQ for 24 h, and DNA adduct formation was analyzed by (32)P-postlabeling. In some experiments, cells were pretreated with the CYP1B1 inhibitor 1-ethynylpyrene (EP) prior to exposure to BP or its metabolites. BP, 3-OH-BP, and BPQ induced formation of several DNA adducts that were not observed in dimethylsulfoxide-treated cells. Re and cochromatography experiments indicated that 3-OH-BP and BPQ were proximate genotoxic metabolites of BP. DNA adduct formation was strongly inhibited by EP, a specific inhibitor of CYP1B1. BP treatment of SMCs resulted in induction of aryl hydrocarbon hydroxylase (AHH) activity and CYP1B1, but not CYP1A1, apoprotein. EP also blocked AHH induction by BP. In conclusion, the results of this study support the hypothesis that in SMCs, which are target sites for the development of atherosclerosis, the major bioactivation pathway of BP entails CYP1B1-mediated formation of the 3-OH-BP and BPQ, which are proximate genotoxic metabolites that may in turn get transformed to ultimate DNA-binding metabolites, which may contribute to atherogenesis by PAHs. PMID- 12649395 TI - Population needs assessment and knee replacement surgery. PMID- 12649396 TI - Fibromyalgia and the therapeutic domain. A philosophical study on the origins of fibromyalgia in a specific social setting. AB - OBJECTIVES: Fibromyalgia has always attracted controversy. Wolfe states that fibromyalgia will always exist regardless of the name given to the syndrome. Hadler describes fibromyalgia as a form of illness behaviour escalated by labelling. However, we believe that fibromyalgia, as other functional somatic syndromes, is not waiting below the surface until it becomes manifest by labelling. METHODS: We developed our hypothesis on the relationship between a specific social setting (called the therapeutic domain) and fibromyalgia using empirical philosophical arguments based on Foucault and Hacking. A therapeutic domain is a real and heterogeneous medical domain in which people, their thoughts and practices, and medical technology in any form coexist and communicate. In this domain blood is aspirated, radiographs are taken and classification criteria are made and applied. It is a domain where patient and therapist have initiated a relationship, which is influenced by the media and political pressure. This results in a looping effect where classification criteria and images give structure to perceptions and form the description for human behaviour; the person thus diagnosed (!) constantly has to grow into the conformity of these classification criteria, which also have to be constantly revised. The fibromyalgia concept becomes manifest in an individual as non-specific aches and pains along with other features. RESULTS: In other times and settings this resulted in analogue syndromes like railway spine, telegraph wrists, neurocirculatory asthenia or perhaps repetitive strain injury. In the application of American College of Rheumatology fibromyalgia classification criteria, labels and medical technology it is possible that invisible experiences manifest themselves in a therapeutic domain. It is not only a phenotype induced by the physician, but in this domain a certain power creates reality making the 'disease' become manifest. CONCLUSION: The only certainty in fibromyalgia is that it is still being diagnosed. For prevention and treatment of fibromyalgia, doctors as well as politicians and media have to start by fundamentally changing the therapeutic domain. In such a renewed setting, fibromyalgia cannot become manifest in an individual and thus fibromyalgia syndrome can no longer exist. A firm public message that symptoms can be psychological in origin to prevent their spread, as Wessely recently stated in the comparable case of mass psychogenic illness, is only a part of the answer. PMID- 12649397 TI - Population requirement for primary knee replacement surgery: a cross-sectional study. AB - OBJECTIVES: To determine the population requirement for total knee replacement (TKR) in England. METHODS: Population-based study using an age/sex-stratified random sample of 28 080 individuals aged 35 yr and over. Incident disease was estimated from prevalence by statistical modelling. The New Zealand priority criteria for major joint replacement were used for case selection. RESULTS: Patients with knee disease were less likely than those with equally severe hip disease to have been referred to a specialist, to have consulted an orthopaedic surgeon or to be on a waiting list for joint replacement. The estimated annual requirement of TKRs in England, based on New Zealand Scores alone, was 55,800 (95% CI 40 700-70,900), contrasting sharply with an annual provision of 29,300 actually observed. However, in contrast to previously reported hip replacement data, when patient willingness to undergo surgery was considered, this estimate decreased considerably. CONCLUSIONS: There appears to be an underprovision of TKR in England. This may be due in part to differences in perception of disease severity and likely response to surgery between patients and general practitioners on one hand, and rheumatologists and orthopaedic surgeons on the other. PMID- 12649398 TI - Elevated levels of synovial fluid antibodies reactive with the small proteoglycans biglycan and decorin in patients with rheumatoid arthritis or other joint diseases. AB - OBJECTIVES: To determine whether patients with rheumatoid arthritis (RA) express humoral immunity to the small proteoglycans biglycan and decorin and to compare the response to that of patients suffering from other joint diseases. METHODS: Serum and synovial fluid IgG and IgM antibody levels were determined by enzyme linked immunosorbent assay. Antibodies to biglycan and decorin as well as to other known and extensively investigated cartilage matrix components such as type II collagen, aggrecan and fibronectin were investigated. Patients suffering from RA, osteoarthritis (OA), psoriatic arthritis and other seronegative spondylarthropathies were included in the study. Correlation between antibody levels and clinical/laboratory parameters was determined. RESULTS: Patients with RA expressed an increased humoral immunity to biglycan, while patients with seronegative spondylarthropathies displayed elevated decorin-specific synovial antibody levels compared with OA patients. CONCLUSION: These results indicate a significantly higher immunity to small proteoglycans in RA and seronegative spondylarthropathies than in OA suggesting a possible involvement in the pathogenesis of inflammatory rheumatic diseases. PMID- 12649399 TI - FcgammaRIIIA-158V and rheumatoid arthritis: a confirmation study. AB - OBJECTIVES: To develop a robust assay for genotyping the FcgammaRIIIA-158V/F polymorphism and to confirm the putative association between the FcgammaRIIIA 158V allele and rheumatoid arthritis (RA). METHODS: This allelic association study examined the FcgammaRIIIA-158V/F polymorphism for association with RA. A novel single-stranded conformational polymorphism assay was used to genotype 828 RA patients and 581 controls from the UK. RESULTS: The FcgammaRIIIA-158V allele was associated with both RA (P=0.02) and nodules (P=0.04). Individuals homozygous for this higher affinity allele had a significantly increased risk of RA (OR 1.53, 95% CI 1.08-2.18) and the development of nodules (OR 2.20, 95% CI 1.20 4.01). There was no evidence of an interaction with the shared epitope. CONCLUSIONS: We have developed a novel assay to genotype the FcgammaRIIIA-158F/V polymorphism and confirmed that homozygosity for the FcgammaRIIIA-158V allele is associated with UK Caucasian RA, particularly in those individuals with nodules, suggesting FcgammaRIIIA may play a role in determining disease severity or in the development of nodules per se. PMID- 12649400 TI - The association of antinuclear antibodies with organ involvement and survival in systemic sclerosis. AB - OBJECTIVES: To study the frequency and specificity of antinuclear antibodies (ANA) and their association with internal organ involvement and survival in systemic sclerosis (SSc). METHODS: Sera from 276 SSc patients were analysed by an indirect immunofluorescence (IIF) technique with HEp-2 cells as a substrate to categorize centromeric (ACA), nucleolar, speckled and homogeneous nuclear IIF patterns. Specific ANA were determined as follows: anti-DNA topoisomerase I (anti topo I) by double immunodiffusion, anti-U1 RNP by passive haemagglutination, anti RNA polymerase I, II and III (anti-RNAP) and anti-histone (AHA) antibodies by enzyme immunoassays. During the follow-up of 7.0+/-4.5 (mean+/-S.D.) yr the occurrence of clinical manifestations and internal organ involvement was registered. RESULTS: ANA were present in 84% of the patients. The most common patterns of the IIF were speckled (41%), homogeneous (25%), nucleolar (24%) and centromeric (18%). A nucleolar pattern was associated with pulmonary fibrosis (P < 0.01) and cardiomegaly (P < 0.05). ACA were related to organic vasculopathy (P < 0.05) and renal involvement (P < 0.01), but not to pulmonary fibrosis (P < 0.01). Anti-topo I were present in 9.4%, anti-U1 RNP in 21%, anti-RNAP in 22% and AHA in 16% of the patients. Pulmonary involvement was more common in patients with anti-topo I (P < 0.05), whereas AHA-positive patients were characterized by cardiac (P < 0.05), pulmonary (P < 0.05) and renal (P < 0.05) involvement. A nucleolar IIF pattern and AHA were both associated with a decreased survival [relative risk of death 1.71 (P < 0.05) and 2.36 (P < 0.01), respectively]. CONCLUSIONS: AHA and a nucleolar HEp-2 cell pattern may indicate critical organ involvement and predict a reduced survival in SSc patients. PMID- 12649401 TI - Influence of therapy with chimeric monoclonal tumour necrosis factor-alpha antibodies on intracellular cytokine profiles of T lymphocytes and monocytes in rheumatoid arthritis patients. AB - INTRODUCTION: It has been shown that T lymphocytes and monocytes/macrophages, producing pro-inflammatory cytokines, play a pivotal role in the pathophysiology of rheumatoid arthritis (RA). In recent placebo-controlled double-blind randomized studies, chimeric (human/mouse) tumour necrosis factor-alpha (TNFalpha) antibodies (cA2) proved to be very effective in improving clinical disease activity and reducing inflammatory parameters in RA. OBJECTIVE: To investigate whether anti-TNFalpha therapy influences the in vitro intracellular cytokine production in peripheral blood monocytes and T lymphocytes of RA patients after one single (24 h) and multiple intravenous infusions (6 months). METHODS: An intracellular flow cytometric technique was applied to measure interleukin 1beta (IL-1beta), IL-6, TNFalpha, IL-10 and IL-12 in monocytes and IL 2, IL-4 and interferon-gamma in T lymphocytes of 15 patients, before, after 24 h and after 6 months of therapy with monoclonal chimeric anti-TNFalpha antibodies (3 mg/kg, bimonthly i.v.). All patients were on stable therapy with methotrexate (15-20 mg/week i.m.). Cytokine content in monocytes was measured directly after blood sampling (basal levels), after 8 h of culture (spontaneous production) and after 8 h of stimulation with lipopolysaccharides (LPS-stimulated production). RESULTS: Basal levels and production (after 8 h) of IL-1beta, IL-6 and TNFalpha were significantly decreased 24 h after the first administration of anti-TNFalpha (for IL-1beta P < 0.01; for IL-6 P < 0.01; for TNF P < 0.003) and after 6 months of therapy (for IL-1beta P < 0.02; for IL-6 P < 0.03; for TNFalpha P < 0.001). For IL-12, basal levels were significantly decreased 24 h and 6 months after the start of therapy with anti-TNFalpha antibodies (P=0.0001; P=0.003, respectively). In contrast, IL-10 production increased significantly after 24 h and after 6 months (P=0.02; P=0.01). The T(H2)/T(H1) cytokine ratio in CD4+ T cells was significantly increased after 24 h and after 6 months of anti-TNFalpha therapy (P=0.003; P=0.0007). CONCLUSION: Anti-TNFalpha therapy might down-regulate the monocytic capacity to produce pro-inflammatory cytokines and induces a shift to a more pronounced anti-inflammatory T(H2) cytokine production. PMID- 12649402 TI - The epidemiology of biopsy-positive giant cell arteritis: special reference to changes in the age of the population. AB - OBJECTIVE: The incidence of giant cell arteritis (GCA) increases with age. The aim of the present study was to investigate whether the increasing incidence of biopsy-proven GCA in Goteborg, Sweden, could be explained in terms of a change in the age composition of the general population. METHODS: All cases of biopsy verified GCA between 1976 and 1995 were recorded. The annual incidence was calculated for women and men aged 50 yr or older and its relationship with the age composition of the general population was tested statistically. RESULTS: There was a significant positive correlation between age and the risk of developing GCA. In the general population, there was a shift towards higher age; in 1976, the mean age of people 50 yr or older was 63.2 (men) and 65.0 (women), whereas in 1995 it was 65.0 (men) and 68.1 (women). After compensating for this, the incidence of biopsy-proven GCA still increased significantly. Moreover, for women aged 50 yr or older, the risk of developing the disease increased more among younger subjects than older ones. CONCLUSIONS: The increase in the incidence of biopsy-proven GCA between 1976 and 1995 could not be explained merely in terms of the increasing age of the general population. It is most probably related to an increase in the influence of other factors. PMID- 12649403 TI - Prospective analysis of the impact of HLA-DR and -DQ on joint destruction in recent-onset rheumatoid arthritis. AB - OBJECTIVE: To evaluate the differential impact of HLA-DR and -DQ on the progression of erosive disease in the clinical course of early rheumatoid arthritis (RA). METHODS: HLA genotyping for HLA-DR and -DQ was carried out in a prospective study of 87 patients with early RA. The progression of erosive disease was assessed by radiological scores over a period of 2 yr in all patients and over 4 yr in 77 patients. The impact of HLA markers was evaluated by univariate comparisons and by multiple logistic regression analyses. RESULTS: Patients expressing the RA-associated shared epitope (SE) on a DRB1*01-positive or, most prominently, on a DRB1*04-positive allele had higher Larsen scores at all time-points analysed when compared with SE-negative patients. A similar impact on radiological progression was seen for the RA-predisposing DQ3, but not for DQ5 heterodimers. In the presence or absence of the DRB1 SE, no additional effects could be discerned for RA-associated DQ molecules. The presence of a DERAA-positive DRB1 allele was associated with a slower pace of joint destruction. While gene dosage effects were seen for SE compound homozygosity, no effect for DQ3 homozygosity could be discerned. CONCLUSION: Although a significant influence of HLA-DQ3 heterodimers on the progression of erosive joint destruction was seen, the analysis of the HLA-DQ locus did not add additional information over the study of HLA-DR including the determination of the SE and the DERAA motif in order to predict the development of severe progressive joint destruction. PMID- 12649404 TI - Response of peripheral blood mononuclear cells from lupus patients to stimulation by CpG oligodeoxynucleotides. AB - OBJECTIVES: Increased levels of hypomethylated CpG-containing DNA in sera from patients with systemic lupus erythematosus (SLE) may contribute to the initiation and/or perpetuation of the disease. This study characterizes the in vitro response of peripheral blood mononuclear cells (PBMC) from SLE patients to CpG DNA. METHODS: Secretion of cytokines and IgM, cell proliferation and up regulation of co-stimulatory molecules were evaluated in PBMC from SLE patients (n=24) and normal controls (n=24) after stimulation with synthetic oligodeoxynucleotides (ODN) containing CpG motifs. RESULTS: Up-regulation of co stimulatory molecules and the secretion of interferon-alpha and interleukin-6 (IL 6) in response to CpG ODN was significantly reduced in monocytes and dendritic cells from SLE patients. Secretion of interferon-gamma by natural killer (NK) cells was also reduced. In contrast, the IgM and IL-10 response of B cells to CpG ODN was normal. CONCLUSION: Monocytes, dendritic cells and NK cells from SLE patients respond abnormally to CpG ODN stimulation, which may contribute to the cytokine imbalance observed in SLE. PMID- 12649405 TI - The three most common CARD15 mutations associated with Crohn's disease and the chromosome 16 susceptibility locus for systemic lupus erythematosus. AB - OBJECTIVE: To test if the three most common mutations contributing to Crohn's disease on the CARD15/NOD2 gene could contribute also to genetic susceptibility to systemic lupus erythematosus (SLE), which has been found to be linked to the region of chromosome 16q13 where the CARD15 gene is located. METHODS: We obtained DNA samples from the blood of 189 SLE patients (according to the American College of Rheumatology classification criteria) and 194 controls of Spanish ancestry. Genotypes for the three CARD15 mutations (3020insC, 2722G>C and 2104C>T) were determined by hybridization with fluorescence resonance energy transfer probes on a LightCycler real-time polymerase chain reaction system. RESULTS: CARD15 genotypes were similar in SLE patients and in controls from the general population (allelic frequencies for 3020insC 0.013 in SLE patients vs 0.013 in controls; for 2722G > C 0.011 vs 0.008; and for 2104C > T 0.032 vs 0.051). CONCLUSION: We did not find evidence that the Crohn's disease-associated mutations on CARD15 contributed to SLE susceptibility. PMID- 12649406 TI - Presentation and management of idiopathic inflammatory muscle disease: four case reports and commentary from a series of 78 patients. PMID- 12649407 TI - Synovial dendritic cells in juvenile idiopathic arthritis (JIA) express receptor activator of NF-kappaB (RANK). AB - OBJECTIVES: To analyse the expression of receptor activator of NF-kappaB (RANK) and RANK ligand (RANKL) in the joints of children with juvenile idiopathic arthritis (JIA), to characterize the phenotype of RANK(+) cells and to test the hypothesis that some RANK(+) cells are of the dendritic type. METHODS: Paired samples of peripheral blood mononuclear cells (PBMC) and synovial fluid mononuclear cells (SFMC) from children with oligoarticular (n=14) or polyarticular (n=4) JIA and PBMC from 10 control subjects were studied for expression of RANK, RANKL and dendritic cell-specific ICAM (intercellular adhesion molecule)-grabbing non-integrin (DC-SIGN) by the reverse transcriptase polymerase chain reaction and three-colour flow cytometry. Expression of DC-SIGN and RANK was followed after 1 week of culture with granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin 4 (IL-4). RESULTS: mRNA for RANK was detected in both adherent cells and T cells from PBMC and SFMC of patients with JIA and in control PBMC, while mRNA for RANKL was detectable in the T-cell fraction from JIA patients but not in that from controls. By flow cytometry, a large number of RANK(+) cells were detected in the joint; these cells had the phenotype HLA-DR(hi)CD86(hi) CD11c(+) and expressed low levels of DC-SIGN. CONCLUSIONS: There is increased expression of RANKL and RANK in the juvenile arthritic joint. RANK is expressed on a population of cells with features of dendritic cells. RANK/RANKL interactions may contribute to the survival of inflammatory cells within the joint, as well as to erosions and osteoporosis in juvenile arthritis. PMID- 12649408 TI - The physiological and physical determinants of functional ability measures in children with juvenile dermatomyositis. AB - OBJECTIVE: To study the relationships of muscle strength and maximal oxygen consumption (VO(2peak)) with Childhood Health Assessment Questionnaire (CHAQ) score, Childhood Myositis Assessment Score (CMAS) and Child Health Questionnaire [physical summary (CHQ-PhS) and psychosocial summary (CHQ-PsS)] scores in juvenile dermatomyositis. METHOD: Fifteen patients (age 5-14 yr) participated. CMAS, CHAQ, CHQ, muscle strength and VO(2peak) were measured. RESULTS: Correlations revealed significant relationships between CHAQ and (i) muscle strength (r=-0.72) and (ii) absolute VO(2peak) (r=-0.68); between CMAS and relative VO(2peak) (r=0.73); and between CHQ-PhS and (i) muscle strength (r=0.57) and (ii) relative VO(2peak) (r=0.58). Backward regression analysis showed that muscle strength was the best indicator of variation in CHAQ. Age and relative VO(2peak) were the best indicators for CMAS. Body mass and age were the best indicators for CHQ-PsS. Body mass and muscle strength were the best indicators for CHQ-PhS. CONCLUSION: CMAS, CHAQ and CHQ correlate with muscle strength and VO(2peak). CMAS, CHAQ and CHQ depend on different physical and physiological variables. PMID- 12649409 TI - Systemic juvenile idiopathic arthritis, Kikuchi's disease and haemophagocytic lymphohistiocytosis--is there a link? Case report and literature review. PMID- 12649410 TI - Severe spongiform aortic stenosis in monozygotic twins with anti-phospholipid syndrome related to systemic lupus erythematosus. PMID- 12649411 TI - Dose reduction of etanercept--can we treat more patients using a fixed budget? PMID- 12649412 TI - Serotonin reuptake inhibitors, Raynaud's phenomenon and erythromelalgia. PMID- 12649413 TI - Nerve conduction studies as a routine diagnostic aid in carpal tunnel syndrome. PMID- 12649414 TI - Survival in fibrosing alveolitis associated with rheumatoid arthritis is better than cryptogenic fibrosing alveolitis. PMID- 12649417 TI - Improved detection of hepatic metastases with pulse-inversion US during the liver specific phase of SHU 508A: multicenter study. AB - PURPOSE: To compare conventional B-mode ultrasonography (US) alone with the combination of conventional B-mode US and contrast material-enhanced (SHU 508A) late-phase pulse-inversion US for the detection of hepatic metastases by using dual-phase spiral computed tomography (CT) as the standard of reference. MATERIALS AND METHODS: One hundred twenty-three patients underwent conventional US, US in the liver-specific phase of SHU 508A, and single-section spiral CT. US and CT images were assessed by blinded readers. Differences in sensitivity, specificity, and the number and smallest size of metastases at conventional and contrast-enhanced US were compared by using CT as the standard of reference. Lesion conspicuity was assessed objectively (quantitatively) and subjectively by one reader before and after contrast material administration. RESULTS: In 45 of 80 (56%) patients with metastases, more metastases were seen at contrast-enhanced US than at conventional US. In three of these patients, conventional US images appeared normal. The addition of contrast-enhanced US improved sensitivity for the detection of individual metastases from 71% to 87% (P <.001). On a patient basis, sensitivity improved from 94% to 98% (P =.44), and specificity improved from 60% to 88% (P <.01). Contrast enhancement improved the subjective conspicuity of metastases in 66 of 75 (88%) patients and the objective contrast by a mean of 10.8 dB (P <.001). Contrast-enhanced US showed more metastases than did CT in seven patients, and CT showed more than did contrast-enhanced US in one of 22 patients in whom an independent reference (magnetic resonance imaging, intraoperative US, or pathologic findings) was available. CONCLUSION: Contrast enhanced US improved sensitivity and specificity in the detection of hepatic metastases. PMID- 12649418 TI - Peritoneal leiomyosarcomatosis originating from gastrointestinal leiomyosarcomas: CT features. AB - PURPOSE: To evaluate the computed tomographic (CT) features of peritoneal leiomyosarcomatosis (PL) originating from gastrointestinal leiomyosarcoma and compare the CT features of primary gastrointestinal leiomyosarcomas between patients with and those without PL. MATERIALS AND METHODS: The authors reviewed the medical and surgical-pathology records and the CT scans of 89 patients with gastrointestinal leiomyosarcoma to determine the prevalence of PL. Also, the CT scans of the patients with PL were evaluated for the morphologic appearance of peritoneal lesions. The CT features of the primary tumors were compared between the patients with and those without PL. Student t and Fisher exact tests were performed. RESULTS: Metastases were present in 39 patients. PL was seen in 25 patients. Other metastatic sites included liver in 26 patients, lymph nodes in 11, lungs in three, and bone in one. In the 25 patients with PL, CT findings included multiple discrete peritoneal nodules in 22 patients and massive peritoneal lesions in three. Smudged omental infiltration was combined with other findings in 10 patients. The sites of the peritoneal masses were mesentery in 18 patients, omentum in 11, paracolic gutters in 11, pelvic cavity in eight, perihepatic space in three, and perisplenic space in one. Four patients had ascites. There were statistically significant differences in size of primary tumor (mean diameter, 12.5 cm vs 9.2 cm) (P =.01) and prevalence of exophytic growth of primary tumor (P =.02) between the patients with and those without PL. CONCLUSION: CT findings can indicate PL in the differential diagnosis. The prevalence of PL appears to be higher when it originates from large primary tumors. PMID- 12649419 TI - Endovascular revascularization below the knee: 6-month results and predictive value of C-reactive protein level. AB - PURPOSE: To determine the association between pre- and postintervention serum C reactive protein (CRP) levels and 6-month restenosis after endovascular treatment of atherosclerotic lesions in arteries below the knee. MATERIALS AND METHODS: Eighty-nine patients with peripheral arterial disease underwent primary successful percutaneous transluminal angioplasty (PTA) of the distal popliteal, anterior tibial, posterior tibial, and fibular arteries. Six-month patency was evaluated with the ankle brachial index (ABI) and color-coded duplex ultrasonography (US). The association between restenosis and preintervention and 48-hour postintervention CRP levels was assessed with multiple logistic regression analysis. RESULTS: ABI improved from a preintervention median of 0.54 to a postintervention median of 0.75 (P <.001). The primary technical success rate was 94% (100 of 106). In 50 patients, a suboptimal technical result was achieved with 30%-40% residual stenosis at the dilated segment. The median ABI at 6 months was 0.65, and it was inversely correlated with preintervention (r = 0.27, P =.009) and 48-hour postintervention (r = -0.40, P <.001) CRP levels. With duplex US at 6 months, restenosis (> or =50%) occurred in 36 patients. Patients with a preintervention CRP level of 0.23-0.92 mg/dL (2.3-9.2 mg/L) had a 3.7-fold increased adjusted risk for restenosis (P =.05); patients with a preintervention CRP level greater than 0.92 mg/dL (9.2 mg/L) had a 4.7-fold increased adjusted risk (P =.03). Postintervention CRP values greater than 2.42 mg/dL (24.2 mg/L) were associated with a 10.7-fold adjusted risk for restenosis (P =.002). Suboptimal PTA result was the only other parameter associated with an increased risk for restenosis (odds ratio, 3.7; P =.03). CONCLUSION: Pre- and postintervention CRP levels were associated with restenosis after PTA of the distal popliteal and tibioperoneal arteries, which indicates that inflammation plays a crucial role in the pathophysiology of this process. PMID- 12649420 TI - Acute-phase response after stent implantation in the carotid artery: association with 6-month in-stent restenosis. AB - PURPOSE: To determine the prognostic importance of periintervention serum levels of acute-phase reactants in 6-month restenosis after stent implantation in the carotid artery. MATERIALS AND METHODS: One hundred eight consecutive patients with 70% or greater stenosis of the internal carotid artery (ICA) according to the North American Symptomatic Carotid Endarterectomy Trial criteria underwent successful stent implantation in the ICA in a prospective cohort study. Six-month patency was evaluated at color-coded duplex ultrasonography. The association between in-stent restenosis (> or =50%, North American Symptomatic Carotid Endarterectomy Trial criterion) and C-reactive protein (CRP) and serum amyloid A levels at baseline and at 24 and 48 hours after intervention was assessed. RESULTS: Restenosis was found in 15 (14%) patients within 6 months. CRP level at 48 hours, exemplary for the postintervention acute-phase response, was significantly (P =.01) associated with 6-month restenosis by using multiple logistic regression analysis. Recurrent stenosis after prior stent implantation in the carotid artery (odds ratio, 9.2; 95% CI: 1.6, 53.2; P =.01) and residual stenosis of 10%-30% after stent implantation (odds ratio, 9.7; 95% CI: 1.6, 59.3; P =.01) were independent clinical predictors of restenosis. CONCLUSION: Extent of vascular inflammatory response after stent implantation in the carotid artery measured with acute-phase reactants is associated with 6-month patency. Recurrent stenosis after prior stent implantation and initial suboptimal technical result seem to be clinically relevant predictors of postangioplasty outcome. PMID- 12649421 TI - Simulated pulmonary nodules implanted in a dedicated porcine chest phantom: sensitivity of MR imaging for detection. AB - PURPOSE: To evaluate the diagnostic accuracy of common magnetic resonance (MR) imaging sequences for detection of small pulmonary nodules by using a chest phantom and porcine lungs containing simulated lesions. MATERIALS AND METHODS: Fourteen porcine lungs containing 366 porcine myocardial tissue implants were inflated inside a phantom. Two-dimensional (2D) and three-dimensional (3D) gradient-echo (GRE), T2-weighted turbo spin-echo (SE), and T2-weighted single shot SE train MR sequences were performed. Spiral computed tomography (CT) was performed for comparison. Blinded observers read the images and recorded the sizes and locations of visible nodules by consensus. The sensitivity of each imaging method for depicting single nodules of given sizes was calculated. Specificities, positive predictive values (PPVs), and negative predictive values (NPVs) for detection of one or more nodules of various sizes were calculated. RESULTS: Sensitivities of 3D GRE, 2D GRE, T2-weighted turbo SE, and T2-weighted single-shot SE train MR imaging and of CT were 0.50, 0.40, 0.12, 0.00, and 0.55, respectively, for detection of 1.4-mm nodules and 0.88, 0.84, 0.69, 0.06, and 0.96, respectively, for detection of 4.2-mm nodules. The 95% CIs for CT and GRE MR imaging overlapped, but those for turbo SE and single-shot SE train MR imaging differed significantly (P <.05). For detection of nodules larger than 5 mm, all examinations except single-shot SE train MR imaging yielded a specificity, PPV, and NPV of 1.00 each. For detection of nodules smaller than 5 mm, diagnostic accuracy of 3D GRE MR imaging was high: Specificity, PPV, and NPV all were approximately 0.90. Two-dimensional GRE MR imaging results were influenced by false-positive findings: Specificity was 0.64; PPV, 0.74; and NPV, 1.00. CONCLUSION: Common MR imaging sequences such as 3D GRE have high diagnostic accuracy in depicting small pulmonary nodules when artifacts from cardiac and respiratory motion are absent. PMID- 12649423 TI - Tightly winding structure of sequential model peptide for repeated helical region in Samia cynthia ricini silk fibroin studied with solid-state NMR. AB - There are many kinds of silks from silkworms and spiders with different structures and properties, and thus, silks are suitable to study the structure property relationship of fibrous proteins. Silk fibroin from a wild silkworm, Samia cynthia ricini, mainly consists of the repeated similar sequences by about 100 times where there are alternative appearances of the polyalanine (Ala)(12-13) region and the Gly-rich region. In this paper, a sequential model peptide, GGAGGGYGGDGG(A)(12)GGAGDGYGAG, which is a typical sequence of the silk fibroin, was synthesized, and the atomic-level conformations of Gly residues at the N- and C-terminal ends of the polyalanine region were determined as well as that of the central Ala residue using (13)C 2D spin diffusion solid-state nuclear magnetic resonance (NMR) under off-magic angle spinning. In the model peptide with alpha helical conformation, the torsion angle of the central Ala residue, the 19th Ala, was determined to be (phi, psi) = (-60 degrees, -50 degrees ), which was a typical alpha-helical structure, but the torsion angles of two Gly residues, the 12th and 25th Gly residues, which are located at the N- and C-terminal ends of the polyalanine region, were determined to be (phi, psi) = (-70 degrees, -30 degrees ) and (phi, psi) = (-70 degrees, -20 degrees ), respectively. Thus, it was observed that the turns at both ends of polyalanine with alpha-helix conformation in the model peptide are tightly wound. PMID- 12649422 TI - How do helix-helix interactions help determine the folds of membrane proteins? Perspectives from the study of homo-oligomeric helical bundles. AB - The final, structure-determining step in the folding of membrane proteins involves the coalescence of preformed transmembrane helices to form the native tertiary structure. Here, we review recent studies on small peptide and protein systems that are providing quantitative data on the interactions that drive this process. Gel electrophoresis, analytical ultracentrifugation, and fluorescence resonance energy transfer (FRET) are useful methods for examining the assembly of homo-oligomeric transmembrane helical proteins. These methods have been used to study the assembly of the M2 proton channel from influenza A virus, glycophorin, phospholamban, and several designed membrane proteins-all of which have a single transmembrane helix that is sufficient for association into a transmembrane helical bundle. These systems are being studied to determine the relative thermodynamic contributions of van der Waals interactions, conformational entropy, and polar interactions in the stabilization of membrane proteins. Although the database of thermodynamic information is not yet large, a few generalities are beginning to emerge concerning the energetic differences between membrane and water-soluble proteins: the packing of apolar side chains in the interior of helical membrane proteins plays a smaller, but nevertheless significant, role in stabilizing their structure. Polar, hydrogen-bonded interactions occur less frequently, but, nevertheless, they often provide a strong driving force for folding helix-helix pairs in membrane proteins. These studies are laying the groundwork for the design of sequence motifs that dictate the association of membrane helices. PMID- 12649424 TI - Denaturation and reassembly of chaperonin GroEL studied by solution X-ray scattering. AB - We measured the denaturation and reassembly of Escherichia coli chaperonin GroEL using small-angle solution X-ray scattering, which is a powerful technique for studying the overall structure and assembly of a protein in solution. The results of the urea-induced unfolding transition show that GroEL partially dissociates in the presence of more than 2 M urea, cooperatively unfolds at around 3 M urea, and is in a monomeric random coil-like unfolded structure at more than 3.2 M urea. Attempted refolding of the unfolded GroEL monomer by a simple dilution procedure is not successful, leading to formation of aggregates. However, the presence of ammonium sulfate and MgADP allows the fully unfolded GroEL to refold into a structure with the same hydrodynamic dimension, within experimental error, as that of the native GroEL. Moreover, the X-ray scattering profiles of the GroEL thus refolded and the native GroEL are coincident with each other, showing that the refolded GroEL has the same structure and the molecular mass as the native GroEL. These results demonstrate that the fully unfolded GroEL monomer can refold and reassemble into the native tetradecameric structure in the presence of ammonium sulfate and MgADP without ATP hydrolysis and preexisting chaperones. Therefore, GroEL can, in principle, fold and assemble into the native structure according to the intrinsic characteristic of its polypeptide chain, although preexisting GroEL would be important when the GroEL folding takes place under in vivo conditions, in order to avoid misfolding and aggregation. PMID- 12649425 TI - Amphiphilic biopolymers (amphibiopols) as new surfactants for membrane protein solubilization. AB - The aim of this study was to develop new surfactants for membrane protein solubilization, from a natural, biodegradable polymer: the polysaccharide pullulan. A set of amphiphilic pullulans (HMCMPs), differing in hydrophobic modification ratio, charge ratio, and the nature of the hydrophobic chains introduced, were synthesized and tested in solubilization experiments with outer membranes of Pseudomonas fluorescens. The membrane proteins were precipitated, and then resolubilized with various HMCMPs. The decyl alkyl chain (C(10)) was the hydrophobic graft that gave the highest level of solubilization. Decyl alkyl chain-bearing HMCMPs were also able to extract integral membrane proteins from their lipid environment. The best results were obtained with an amphiphilic pullulan bearing 18% decyl groups (18C(10)). Circular dichroism spectroscopy and membrane reconstitution experiments were used to test the structural and functional integrity of 18C(10)-solubilized proteins (OmpF from Escherichia coli and bacteriorhodopsin from Halobacterium halobium). Whatever their structure type (alpha or beta), 18C(10) did not alter either the structure or the function of the proteins analyzed. Thus, HMCMPs appear to constitute a promising new class of polymeric surfactants for membrane protein studies. PMID- 12649426 TI - Stable octameric structure of recombinant hemoglobin alpha(2)beta(2)83 Gly-->Cys. AB - We have engineered a recombinant hemoglobin (rHb betaG83C) based on the variant Hb Ta-Li, which oligomerizes through intertetramer disulfide bonds. Size exclusion chromatography and electrospray ionization mass spectrometry show that the rHb betaG83C assembles into an oligomeric structure the size of a dimer of tetramers. The oligomer has carbon monoxide-binding properties similar to those of natural human hemoglobin. Unlike HbA, the oligomer does not participate in dimer exchange. The CO kinetics, auto-oxidation rate, and gel filtration experiments on the oligomeric betaG83C did not show the usual concentration dependence, implying that it does not dissociate easily into smaller species. The octamer could be dissociated by the use of reducing agents. The action of reduced glutathione on oligomeric betaG83C exhibited biphasic kinetics for the loss of the octameric form, with a time constant for the rapid phase of about 2 h at 1 mM glutathione. However, the size of oligomer betaG83C was not modified after incubation with fresh plasma. PMID- 12649427 TI - Prediction of protein domain boundaries from sequence alone. AB - We present here a simple approach to identify domain boundaries in proteins of an unknown three-dimensional structure. Our method is based on the hypothesis that a high-side chain entropy of a region in a protein chain must be compensated by a high-residue interaction energy within the region, which could correlate with a well-structured part of the globule, that is, with a domain unit. For protein domains, this means that the domain boundary is conditioned by amino acid residues with a small value of side chain entropy, which correlates with the side chain size. On the one hand, relatively high Ala and Gly content on the domain boundary results in high conformational entropy of the backbone chain between the domains. On the other hand, the presence of Pro residues leads to the formation of hinges for a relative orientation of domains. The method was applied to 646 proteins with two contiguous domains extracted from the SCOP database with a success rate of 63%. We also report the prediction of domain boundaries for CASP5 targets obtained with the same method. PMID- 12649428 TI - Polycation-induced oligomerization and accelerated fibrillation of human alpha synuclein in vitro. AB - The aggregation and fibrillation of alpha-synuclein has been implicated as a causative factor in Parkinson's disease and several other neurodegenerative disorders known as synucleinopathies. The effect of different factors on the process of fibril formation has been intensively studied in vitro. We show here that alpha-synuclein interacts with different unstructured polycations (spermine, polylysine, polyarginine, and polyethyleneimine) to form specific complexes. In addition, the polycations catalyze alpha-synuclein oligomerization. The formation of alpha-synuclein-polycation complexes was not accompanied by significant structural changes in alpha-synuclein. However, alpha-synuclein fibrillation was dramatically accelerated in the presence of polycations. The magnitude of the accelerating effect depended on the nature of the polymer, its length, and concentration. The results illustrate the potential critical role of electrostatic interactions in protein aggregation, and the potential role of naturally occurring polycations in modulating alpha-synuclein aggregation. PMID- 12649429 TI - The likelihood of aggregation during protein renaturation can be assessed using the second virial coefficient. AB - Protein aggregation is commonly observed during protein refolding. To better understand this phenomenon, the intermolecular interactions experienced by a protein during unfolding and refolding are inferred from second virial coefficient (SVC) measurements. It is accepted that a negative SVC is indicative of protein-protein interactions that are attractive, whereas a positive SVC indicates net repulsive interactions. Lysozyme denatured and reduced in guanidinium hydrochloride exhibited a decreasing SVC as the denaturant was diluted, and the SVC approached zero at approximately 3 M GdnHCl. Further dilution of denaturant to renaturation conditions (1.25 M GdnHCl) led to a negative SVC, and significant protein aggregation was observed. The inclusion of 500 mM L-arginine in the renaturation buffer shifted the SVC to positive and suppressed aggregation, thereby increasing refolding yield. The formation of mixed disulfides in the denatured state prior to refolding also increased protein solubility and suppressed aggregation, even without the use of L-arginine. Again, the suppression of aggregation was shown to be caused by a shift from attractive to repulsive intermolecular interactions as reflected in a shift from a negative to a positive SVC value. To the best of our knowledge, this is the first time that SVC data have been reported for renaturation studies. We believe this technique will aid in our understanding of how certain conditions promote renaturation and increase protein solubility, thereby suppressing aggregation. SVC measurements provide a useful link, for protein folding and aggregation, between empirical observation and thermodynamics. PMID- 12649430 TI - Structural dissection of alkaline-denatured pepsin. AB - It has been established in a number of studies that the alkaline-denatured state of pepsin (the I(P) state) is composed of a compact C-terminal lobe and a largely unstructured N-terminal lobe. In the present study, we have investigated the residual structure in the I(P) state in more detail, using limited proteolysis to isolate and characterize a tightly folded core region from this partially denatured pepsin. The isolated core region corresponds to the 141 C-terminal residues of the pepsin molecule, which in the fully native state forms one of the two lobes of the structure. A comparative study using NMR and CD spectroscopy has revealed, however, that the N-terminal lobe contributes a substantial amount of additional residual structure to the I(P) state of pepsin. CD spectra indicate in addition that significant nonnative alpha-helical structure is present in the C terminal lobe of the structure when the N-terminal lobe of pepsin is either unfolded or removed by proteolysis. This study demonstrates that the structure of pepsin in the I(P) state is significantly more complex than that of a fully folded C-terminal lobe connected to an unstructured N-terminal lobe. PMID- 12649431 TI - Using nondenaturing mass spectrometry to detect fortuitous ligands in orphan nuclear receptors. AB - Nondenaturing electrospray mass spectrometry (ESI-MS) has been used to reveal the presence of potential ligands in the ligand-binding domain (LBD) of orphan nuclear receptors. This new approach, based on supramolecular mass spectrometry, allowed the detection and identification of fortuitous ligands for the retinoic acid-related orphan receptor beta (RORbeta) and the ultraspiracle protein (USP). These fortuitous ligands were specifically captured from the host cell with the proper stoichiometry. After organic extraction, these molecules have been characterized by classic analytical methods and identified as stearic acid for RORbeta and a phosphatidylethanolamine (PE) for USP, as confirmed by crystallography. These molecules act as "fillers" and may not be the physiological ligands, but they prove to be essential to stabilize the active conformation of the LBD, enabling its crystallization. The resulting crystal structures provide a detailed picture of the ligand-binding pocket, allowing the design of highly specific synthetic ligands that can be used to characterize the function of orphan nuclear receptors. An additional advantage of this new method is that it is not based on a functional test and that it can detect low-affinity ligands. PMID- 12649432 TI - Noncovalent scFv multimers of tumor-targeting anti-Lewis(y) hu3S193 humanized antibody. AB - Single-chain variable fragments (scFvs) of anti-Lewis(y) hu3S193 humanized antibody were constructed by joining the V(H) and V(L) domains with either +2 residues, +1 residue, or by directly linking the domains. In addition two constructs were synthesized in which one or two C-terminal residues of the V(H) domain were removed (-1 residue, -2 residue) and then joined directly to the V(L) domain. An scFv construct in the reverse orientation with the V(L) joined directly to the V(H) domain was also synthesized. Upon transformation into Escherichia coli all scFv constructs expressed active protein. Binding activity, multimeric status, and multivalent properties were assessed by flow cytometry, size exclusion chromatography, and biosensor analysis. The results for hu3S193 scFvs are consistent with the paradigm that scFvs with a linker of +3 residues or more associate to form a non-covalent dimer, and those with a shorter linker or directly linked associate predominantly to form a non-covalent trimer and tetramer that are in equilibrium. While the association of V domains to form either a dimer or trimer/tetramer is governed by the length of the linker, the stability of the trimer/tetramer in the equilibrium mixture is dependent on the affinity of the interaction of the individual V domains to associate to form the larger Fv module. PMID- 12649433 TI - Prediction of the plant beta-barrel proteome: a case study of the chloroplast outer envelope. AB - In the postgenomic era, the transformation of genetic information into biochemical meaning is required. We have analyzed the proteome of the chloroplast outer envelope membrane by an in silico and a proteomic approach. Based on its evolutionary relation to the outer membrane of Gram-negative bacteria, the outer envelope membrane should contain a large number of beta-barrel proteins. We therefore calculated the probability for the existence of beta-sheet, beta barrel, and hairpin structures among all proteins of the Arabidopsis thaliana genome. According to the existence of these structures, a number of candidates were selected. This protein pool was analyzed by TargetP to discard sequences with signals that would direct the protein to other organelles different from chloroplasts. In addition, the pool was manually controlled for the presence of proteins known to function outside of the chloroplast envelope. The approach developed here can be used to predict the topology of beta-barrel proteins. For the proteomic approach, proteins of highly purified outer envelope membranes of chloroplasts from Pisum sativum were analyzed by ESI-MS/MS mass spectrometry. In addition to the known components, four new proteins of the outer envelope membranes were identified in this study. PMID- 12649434 TI - Unexpected similarity in regulation between an archaeal inositol monophosphatase/fructose bisphosphatase and chloroplast fructose bisphosphatase. AB - Hyperthermophilic archaea have an unusual phosphatase that exhibits activity toward both inositol-1-phosphate and fructose-1,6-bisphosphate, activities carried out by separate gene products in eukaryotes and bacteria. The structures of phosphatases from Archaeoglobus fulgidus (AF2372) and Methanococcus jannaschii (MJ0109), both anaerobic organisms, resemble the dimeric unit of the tetrameric pig kidney fructose bisphosphatase (FBPase). A striking feature of AF2372, but not of MJ0109, is that the sulfhydryl groups of two cysteines, Cys150 and Cys186, are in close proximity (4 A). A similar arrangement of cysteines has been observed in chloroplast FBPases that are regulated by disulfide formation controlled by redox signaling pathways (ferredoxin/thioredoxin). This mode of regulation has not been detected in any other FBPase enzymes. Biochemical assays show that the AF2372 phosphatase activity can be abolished by incubation with O(2). Full activity is restored by incubation with thiol-containing compounds. Neither the C150S variant of AF2372 nor the equivalent phosphatase from M. jannaschii loses activity with oxidation. Oxidation experiments using Escherichia coli thioredoxin, in analogy with the chloroplast FBPase system, indicate an unexpected mode of regulation for AF2372, a key phosphatase in this anaerobic sulfate reducer. PMID- 12649435 TI - A peptide model of insulin folding intermediate with one disulfide. AB - Insulin folds into a unique three-dimensional structure stabilized by three disulfide bonds. Our previous work suggested that during in vitro refolding of a recombinant single-chain insulin (PIP) there exists a critical folding intermediate containing the single disulfide A20-B19. However, the intermediate cannot be trapped during refolding because once this disulfide is formed, the remaining folding process is very quick. To circumvent this difficulty, a model peptide ([A20-B19]PIP) containing the single disulfide A20-B19 was prepared by protein engineering. The model peptide can be secreted from transformed yeast cells, but its secretion yield decreases 2-3 magnitudes compared with that of the wild-type PIP. The physicochemical property analysis suggested that the model peptide adopts a partially folded conformation. In vitro, the fully reduced model peptide can quickly and efficiently form the disulfide A20-B19, which suggested that formation of the disulfide A20-B19 is kinetically preferred. In redox buffer, the model peptide is reduced gradually as the reduction potential is increased, while the disulfides of the wild-type PIP are reduced in a cooperative manner. By analysis of the model peptide, it is possible to deduce the properties of the critical folding intermediate with the single disulfide A20-B19. PMID- 12649436 TI - Structural and kinetic characterization of the simplified SH3 domain FP1. AB - The simplified SH3 domain sequence, FP1, obtained in phage display selection experiments has an amino acid composition that is 95% Ile, Lys, Glu, Ala, Gly. Here we use NMR to investigate the tertiary structure of FP1. We find that the overall topology of FP1 resembles that of the src SH3 domain, the hydrogen deuterium exchange and chemical shift perturbation profiles are similar to those of naturally occurring SH3 domains, and the (15)N relaxation rates are in the range of naturally occurring small proteins. Guided by the structure, we further simplify the FP1 sequence and compare the effects on folding kinetics of point mutations in FP1 and the wild-type src SH3 domain. The results suggest that the folding transition state of FP1 is similar to but somewhat less polarized than that of the wild-type src SH3 domain. PMID- 12649437 TI - Rational design, synthesis, and verification of affinity ligands to a protein surface cleft. AB - The structure-based design, synthesis, and screening of a glucuronic acid scaffold library of affinity ligands directed toward the catalytic cleft on porcine pancreas alpha-amylase are presented. The design was based on the simulated docking to the enzyme active site of 53 aryl glycosides from the Available Chemicals Directory (ACD) selected by in silico screening. Twenty-three compounds were selected for synthesis and screened in solution for binding toward alpha-amylase using nuclear magnetic resonance techniques. The designed molecules include a handle outside of the binding site to allow their attachment to various surfaces with minimal loss of binding activity. After initial screening in solution, one affinity ligand was selected, immobilized to Sepharose (Amersham Biosciences), and evaluated as a chromatographic probe. A column packed with ligand-coupled Sepharose specifically retained the enzyme, which could be eluted by a known inhibitor. PMID- 12649438 TI - Tautomerism, acid-base equilibria, and H-bonding of the six histidines in subtilisin BPN' by NMR. AB - We have determined by (15)N, (1)H, and (13)C NMR, the chemical behavior of the six histidines in subtilisin BPN' and their PMSF and peptide boronic acid complexes in aqueous solution as a function of pH in the range of from 5 to 11, and have assigned every (15)N, (1)H, C(epsilon 1), and C(delta2) resonance of all His side chains in resting enzyme. Four of the six histidine residues (17, 39, 67, and 226) are neutrally charged and do not titrate. One histidine (238), located on the protein surface, titrates with pK(a) = 7.30 +/- 0.03 at 25 degrees C, having rapid proton exchange, but restricted mobility. The active site histidine (64) in mutant N155A titrates with a pK(a) value of 7.9 +/- 0.3 and sluggish proton exchange behavior, as shown by two-site exchange computer lineshape simulation. His 64 in resting enzyme contains an extremely high C(epsilon 1)-H proton chemical shift of 9.30 parts per million (ppm) owing to a conserved C(epsilon 1)-H(.)O=C H-bond from the active site imidazole to a backbone carbonyl group, which is found in all known serine proteases representing all four superfamilies. Only His 226, and His 64 at high pH, exist as the rare N(delta1)-H tautomer, exhibiting (13)C(delta1) chemical shifts approximately 9 ppm higher than those for N(epsilon 2)-H tautomers. His 64 in the PMSF complex, unlike that in the resting enzyme, is highly mobile in its low pH form, as shown by (15)N-(1)H NOE effects, and titrates with rapid proton exchange kinetics linked to a pK(a) value of 7.47 +/- 0.02. PMID- 12649439 TI - NMR characterization of interleukin-2 in complexes with the IL-2Ralpha receptor component, and with low molecular weight compounds that inhibit the IL-2/IL Ralpha interaction. AB - Nuclear magnetic resonance (NMR) methods were employed to study the interaction of the cytokine Interleukin-2 (IL-2) with the alpha-subunit of its receptor (IL 2Ralpha), and to help understand the behavior of small molecule inhibitors of this interaction. Heteronuclear (1)H-(15)N HSQC experiments were used to identify the interaction surface of (15)N-enriched Interleukin-2 ((15)N-IL-2) in complex with human IL-2Ralpha. In these experiments, chemical shift and line width changes in the heteronuclear single-quantum coherence (HSQC) spectra upon binding of (15)N-IL-2 enabled classification of NH atoms as either near to, or far from, the IL-2Ralpha interaction surface. These data were complemented by hydrogen/deuterium (H/D) exchange measurements, which illustrated enhanced protection of slowly-exchanging IL-2 NH protons near the site of interaction with IL-2Ralpha. The interaction surface defined by NMR compared well with the IL 2Ralpha binding site identified previously using mutagenesis of human and murine IL-2. Two low molecular weight inhibitors of the IL-2/IL-2Ralpha interaction were studied: one (a cyclic peptide derivative) was found to mimic a part of the cytokine and bind to IL-2Ralpha; the other (an acylphenylalanine derivative) was found to bind to IL-2. For the interaction between IL-2 and the acylphenylalanine, chemical shift perturbations of (15)N and (15)NH backbone resonances were tracked as a function of ligand concentration. The perturbation pattern observed for this complex revealed that the acylphenylalanine is a competitive inhibitor-it binds to the same site on IL-2 that interacts with IL 2Ralpha. PMID- 12649440 TI - Intermolecular ion pairs maintain the toroidal structure of Pyrococcus furiosus PCNA. AB - Two mutant proliferating cell nuclear antigens from the hyperthermophilic archaeon Pyrococcus furiosus, PfuPCNA(D143A) and PfuPCNA(D143A/D147A), were prepared by site-specific mutagenesis. The results from gel filtration showed that mutations at D143 and D147 drastically affect the stability of the trimeric structure of PfuPCNA. The PfuPCNA(D143A) still retained the activity to stimulate the DNA polymerase reaction, but PfuPCNA(D143A/D147A) lost the activity. Crystal structures of the mutant PfuPCNAs were determined. Although the wild-type PCNA forms a toroidal trimer with intermolecular hydrogen bonds between the N- and C terminal domains, the mutant PfuPCNAs exist as V-shaped dimers through intermolecular hydrogen bonds between the two C-terminal domains in the crystal. Because the mutated residues are involved in the intermolecular ion pairs through their side chains in the wild-type PfuPCNA, these ion pairs seem to play a key role in maintaining the toroidal structure of the PfuPCNA trimer. The comparison of the crystal structures revealed intriguing conformational flexibility of each domain in the PfuPCNA subunit. This structural versatility of PCNA may be involved in the mechanisms for ring opening and closing. PMID- 12649441 TI - The oligomerization and ligand-binding properties of Sm-like archaeal proteins (SmAPs). AB - Intron splicing is a prime example of the many types of RNA processing catalyzed by small nuclear ribonucleoprotein (snRNP) complexes. Sm proteins form the cores of most snRNPs, and thus to learn principles of snRNP assembly we characterized the oligomerization and ligand-binding properties of Sm-like archaeal proteins (SmAPs) from Pyrobaculum aerophilum (Pae) and Methanobacterium thermautotrophicum (Mth). Ultracentrifugation shows that Mth SmAP1 is exclusively heptameric in solution, whereas Pae SmAP1 forms either disulfide-bonded 14-mers or sub heptameric states (depending on the redox potential). By electron microscopy, we show that Pae and Mth SmAP1 polymerize into bundles of well ordered fibers that probably form by head-to-tail stacking of heptamers. The crystallographic results reported here corroborate these findings by showing heptamers and 14-mers of both Mth and Pae SmAP1 in four new crystal forms. The 1.9 A-resolution structure of Mth SmAP1 bound to uridine-5'-monophosphate (UMP) reveals conserved ligand binding sites. The likely RNA binding site in Mth agrees with that determined for Archaeoglobus fulgidus (Afu) SmAP1. Finally, we found that both Pae and Mth SmAP1 gel-shift negatively supercoiled DNA. These results distinguish SmAPs from eukaryotic Sm proteins and suggest that SmAPs have a generic single-stranded nucleic acid-binding activity. PMID- 12649442 TI - Comparison of family 12 glycoside hydrolases and recruited substitutions important for thermal stability. AB - As part of a program to discover improved glycoside hydrolase family 12 (GH 12) endoglucanases, we have studied the biochemical diversity of several GH 12 homologs. The H. schweinitzii Cel12A enzyme differs from the T. reesei Cel12A enzyme by only 14 amino acids (93% sequence identity), but is much less thermally stable. The bacterial Cel12A enzyme from S. sp. 11AG8 shares only 28% sequence identity to the T. reesei enzyme, and is much more thermally stable. Each of the 14 sequence differences from H. schweinitzii Cel12A were introduced in T. reesei Cel12A to determine the effect of these amino acid substitutions on enzyme stability. Several of the T. reesei Cel12A variants were found to have increased stability, and the differences in apparent midpoint of thermal denaturation (T(m)) ranged from a 2.5 degrees C increase to a 4.0 degrees C decrease. The least stable recruitment from H. schweinitzii Cel12A was A35S. Consequently, the A35V substitution was recruited from the more stable S. sp. 11AG8 Cel12A and this T. reesei Cel12A variant was found to have a T(m) 7.7 degrees C higher than wild type. Thus, the buried residue at position 35 was shown to be of critical importance for thermal stability in this structural family. There was a ninefold range in the specific activities of the Cel12 homologs on o-NPC. The most and least stable T. reesei Cel12A variants, A35V and A35S, respectively, were fully active. Because of their thermal tolerance, S. sp. 11AG8 Cel12A and T. reesei Cel12A variant A35V showed a continual increase in activity over the temperature range of 25 degrees C to 60 degrees C, whereas the less stable enzymes T. reesei Cel12A wild type and the destabilized A35S variant, and H. schweinitzii Cel12A showed a decrease in activity at the highest temperatures. The crystal structures of the H. schweinitzii, S. sp. 11AG8, and T. reesei A35V Cel12A enzymes have been determined and compared with the wild-type T. reesei Cel12A enzyme. All of the structures have similar Calpha traces, but provide detailed insight into the nature of the stability differences. These results are an example of the power of homolog recruitment as a method for identifying residues important for stability. PMID- 12649443 TI - Structure and dynamics of the DNA-binding protein HU of B. stearothermophilus investigated by Raman and ultraviolet-resonance Raman spectroscopy. AB - The histone-like protein HU of Bacillus stearothermophilus (HUBst) is a 90 residue homodimer that binds nonspecifically to B DNA. Although the structure of the HUBst:DNA complex is not known, the proposed DNA-binding surface consists of extended arms that project from an alpha-helical platform. Here, we report Raman and ultraviolet-resonance Raman (UVRR) spectra diagnostic of subunit secondary structures and indicative of key side-chains lining the proposed DNA-binding surface. Raman conformation markers show that the DNA-binding arms of the dimer contain beta-stranded structure in excess (eight +/- two residues per subunit) of that reported previously. Important among side-chain markers are Met (701 cm( 1)), Ala (908 cm(-1)), Arg (1082 cm(-1)), and Pro (1457 cm(-1)). The Ala marker undergoes a substantial shift (908 --> 893 cm(-1)) on deuteration of alanyl peptide sites, indicating a coupled side-chain/main-chain mode of diagnostic value in the identification of exchange-protected alanines. A large subset of alanines (67%) in the alpha-helical core exhibits robust resistance to exchange. A quantitative study of NH --> ND exchange exploiting newly identified amide II' markers of helical (1440 cm(-1)) and nonhelical (1472 cm(-1)) conformations of HUBst indicates unexpected flexibility at the dimer interface, which is manifested in rapid exchange of 80% of peptide sites. The results establish a basis for subsequent Raman and UVRR investigations of HUBst:DNA complexes and provide a framework for applications to other DNA-binding architectural proteins. PMID- 12649444 TI - Anion binding to a protein-protein complex lacks dependence on net charge. AB - The binding of anions to proteins occurs in numerous physiological and metabolic processes. In an effort to understand the factors important in these interactions, we have studied the weak binding of phosphate and sulfate to a protein-protein complex using isothermal titration calorimetry. To our knowledge, this is the first system in which the thermodynamics of anion binding have been determined calorimetrically. By studying both phosphate and sulfate binding and using a range of pH values, the charge on the anion was varied from approximately -1 to -2. Surprisingly, no dependence of the binding energetics on the charge of the anion was observed. This result indicates that charge-charge interactions are not the dominant factor in binding and suggests the importance of hydrogen bonding in specifically recognizing and coordinating anions. PMID- 12649446 TI - The climate divide. PMID- 12649445 TI - Analyses of circular dichroism spectra of membrane proteins. AB - Circular dichroism (CD) spectroscopy is a valuable technique for the determination of protein secondary structures. Many linear and nonlinear algorithms have been developed for the empirical analysis of CD data, using reference databases derived from proteins of known structures. To date, the reference databases used by the various algorithms have all been derived from the spectra of soluble proteins. When applied to the analysis of soluble protein spectra, these methods generally produce calculated secondary structures that correspond well with crystallographic structures. In this study, however, it was shown that when applied to membrane protein spectra, the resulting calculations produce considerably poorer results. One source of this discrepancy may be the altered spectral peak positions (wavelength shifts) of membrane proteins due to the different dielectric of the membrane environment relative to that of water. These results have important consequences for studies that seek to use the existing soluble protein reference databases for the analyses of membrane proteins. PMID- 12649447 TI - Infectious diseases. Scientists chase fast-moving and deadly global illness. PMID- 12649448 TI - Federal work force. Can outsiders do better in managing NIH grants? PMID- 12649449 TI - France. New cuts slam most research agencies. PMID- 12649451 TI - Math and science education. Congress lets states share the wealth. PMID- 12649450 TI - West Nile virus. Blood banks in a 'race against the mosquitoes'. PMID- 12649452 TI - Agrobiotechnology. Foundation goes to bat for African farmers. PMID- 12649453 TI - State-based initiatives. Rising deficits could doom science programs. PMID- 12649454 TI - NASA science probe. Gravity mission faces make-or-break review. PMID- 12649455 TI - Environmental health. Academy to mediate debate over rocket-fuel contaminants. PMID- 12649456 TI - Biotechnology. Stem cells lose market luster. PMID- 12649457 TI - Graduate education. New Asian schools look west for high-quality instruction. PMID- 12649458 TI - Astrophysics. Robotic telescopes catch up with gamma ray bursts. PMID- 12649459 TI - Conservation. Unwelcome bison may face a DNA test. PMID- 12649460 TI - Economics. How much are human lives and health worth? PMID- 12649461 TI - Economics. Wielding the data-quality cudgel. PMID- 12649462 TI - A risky forest policy in the Amazon? PMID- 12649463 TI - The EU and the dinosaur. PMID- 12649464 TI - Assessing environmental changes in grasslands. PMID- 12649465 TI - Comment on "The influence of the proinflammatory cytokine, osteopontin, on autoimmune demyelinating disease". PMID- 12649467 TI - Conservation. Fate of Iraqi archaeology. PMID- 12649468 TI - Physics. Flux qubit completes the hat trick. PMID- 12649469 TI - Anthropology. Overkill and sustainable use. PMID- 12649470 TI - Botany. Relieving DELLA restraint. PMID- 12649471 TI - Evolution. Wingless insects and plucked chickens. PMID- 12649472 TI - Geophysics. Sandbars in motion. PMID- 12649473 TI - Neuroscience. Gambling on dopamine. PMID- 12649474 TI - Signaling life and death in the thymus: timing is everything. AB - T lymphocytes are generated in the thymus, where developing thymocytes must accept one of two fates: They either differentiate or they die. These fates are chiefly determined by signals that originate from the T cell receptor (TCR), a single receptor complex with a remarkable capacity to decide between distinct cell fates. This review explores TCR signaling in thymocytes and focuses on the kinetic aspects of ligand binding, coreceptor involvement, protein phosphorylation, and mitogen-activated protein kinase (MAPK) activation. Understanding the logic of TCR signaling may eventually explain how thymocytes and T cells distinguish self from nonself, a phenomenon that has fascinated immunologists for 50 years. PMID- 12649475 TI - Imperceptible stimuli and sensory processing impediment. PMID- 12649476 TI - Templating organic semiconductors via self-assembly of polymer colloids. AB - A route for producing semiconducting polymer blends is demonstrated in which a doped pi-conjugated polymer is forced into a three-dimensionally continuous minor phase by the self-assembly of colloidal particles and block copolymers. The resulting cellular morphology can be viewed as a high-internal phase polymeric emulsion. Compared with traditional blending procedures, this process reduces the percolation threshold for electrical conductivity by a factor of 10, increases the conductivity by several orders of magnitude, and simultaneously improves thermal stability. Following this route, new applications can be envisaged for semiconducting polymer blends that require only minimal concentrations of doped pi-conjugated polymer. PMID- 12649477 TI - "Plugging into Enzymes": nanowiring of redox enzymes by a gold nanoparticle. AB - The reconstitution of an apo-flavoenzyme, apo-glucose oxidase, on a 1.4-nanometer gold nanocrystal functionalized with the cofactor flavin adenine dinucleotide and integrated into a conductive film yields a bioelectrocatalytic system with exceptional electrical contact with the electrode support. The electron transfer turnover rate of the reconstituted bioelectrocatalyst is approximately 5000 per second, compared with the rate at which molecular oxygen, the natural cosubstrate of the enzyme, accepts electrons (approximately 700 per second). The gold nanoparticle acts as an electron relay or "electrical nanoplug" for the alignment of the enzyme on the conductive support and for the electrical wiring of its redox-active center. PMID- 12649478 TI - Self-aligned, vertical-channel, polymer field-effect transistors. AB - The manufacture of high-performance, conjugated polymer transistor circuits on flexible plastic substrates requires patterning techniques that are capable of defining critical features with submicrometer resolution. We used solid-state embossing to produce polymer field-effect transistors with submicrometer critical features in planar and vertical configurations. Embossing is used for the controlled microcutting of vertical sidewalls into polymer multilayer structures without smearing. Vertical-channel polymer field-effect transistors on flexible poly(ethylene terephthalate) substrates were fabricated, in which the critical channel length of 0.7 to 0.9 micrometers was defined by the thickness of a spin coated insulator layer. Gate electrodes were self-aligned to minimize overlap capacitance by inkjet printing that used the embossed grooves to define a surface energy pattern. PMID- 12649479 TI - Wave-induced sediment transport and sandbar migration. AB - Onshore sediment transport and sandbar migration are important to the morphological evolution of beaches but are not well understood. Here, a model that accounts for fluid accelerations in waves predicts the onshore sandbar migration observed on an ocean beach. In both the observations and the model, the location of the maximum acceleration-induced transport moves shoreward with the sandbar, resulting in feedback between waves and morphology that drives the bar shoreward until conditions change. A model that combines the effects of transport by waves and mean currents simulated both onshore and offshore bar migration observed over a 45-day period. PMID- 12649480 TI - Hexapod origins: monophyletic or paraphyletic? AB - Recent morphological and molecular evidence has changed interpretations of arthropod phylogeny and evolution. Here we compare complete mitochondrial genomes to show that Collembola, a wingless group traditionally considered as basal to all insects, appears instead to constitute a separate evolutionary lineage that branched much earlier than the separation of many crustaceans and insects and independently adapted to life on land. Therefore, the taxon Hexapoda, as commonly defined to include all six-legged arthropods, is not monophyletic. PMID- 12649481 TI - Role of EphA4 and EphrinB3 in local neuronal circuits that control walking. AB - Local circuits in the spinal cord that generate locomotion are termed central pattern generators (CPGs). These provide coordinated bilateral control over the normal limb alternation that underlies walking. The molecules that organize the mammalian CPG are unknown. Isolated spinal cords from mice lacking either the EphA4 receptor or its ligand ephrinB3 have lost left-right limb alternation and instead exhibit synchrony. We identified EphA4-positive neurons as an excitatory component of the locomotor CPG. Our study shows that dramatic locomotor changes can occur as a consequence of local genetic rewiring and identifies genes required for the development of normal locomotor behavior. PMID- 12649482 TI - Identifying kinetic barriers to mechanical unfolding of the T. thermophila ribozyme. AB - Mechanical unfolding trajectories for single molecules of the Tetrahymena thermophila ribozyme display eight intermediates corresponding to discrete kinetic barriers that oppose mechanical unfolding with lifetimes of seconds and rupture forces between 10 and 30 piconewtons. Barriers are magnesium dependent and correspond to known intra- and interdomain interactions. Several barrier structures are "brittle," breakage requiring high forces but small (1 to 3 nanometers) deformations. Barrier crossing is stochastic, leading to variable unfolding paths. The response of complex RNA structures to locally applied mechanical forces may be analogous to the responses of RNA during translation, messenger RNA export from the nucleus, and viral replication. PMID- 12649483 TI - Accumulation of phosphorylated repressor for gibberellin signaling in an F-box mutant. AB - Gibberellin (GA) regulates growth and development in plants. We isolated and characterized a rice GA-insensitive dwarf mutant, gid2. The GID2 gene encodes a putative F-box protein, which interacted with the rice Skp1 homolog in a yeast two-hybrid assay. In gid2, a repressor for GA signaling, SLR1, was highly accumulated in a phosphorylated form and GA increased its concentration, whereas SLR1 was rapidly degraded by GA through ubiquitination in the wild type. We conclude that GID2 is a positive regulator of GA signaling and that regulated degradation of SLR1 is initiated through GA-dependent phosphorylation and finalized by an SCF(GID2)-proteasome pathway. PMID- 12649484 TI - Discrete coding of reward probability and uncertainty by dopamine neurons. AB - Uncertainty is critical in the measure of information and in assessing the accuracy of predictions. It is determined by probability P, being maximal at P = 0.5 and decreasing at higher and lower probabilities. Using distinct stimuli to indicate the probability of reward, we found that the phasic activation of dopamine neurons varied monotonically across the full range of probabilities, supporting past claims that this response codes the discrepancy between predicted and actual reward. In contrast, a previously unobserved response covaried with uncertainty and consisted of a gradual increase in activity until the potential time of reward. The coding of uncertainty suggests a possible role for dopamine signals in attention-based learning and risk-taking behavior. PMID- 12649485 TI - Identified sources and targets of slow inhibition in the neocortex. AB - There are two types of inhibitory postsynaptic potentials in the cerebral cortex. Fast inhibition is mediated by ionotropic gamma-aminobutyric acid type A (GABA(A)) receptors, and slow inhibition is due to metabotropic GABA(B) receptors. Several neuron classes elicit inhibitory postsynaptic potentials through GABA(A) receptors, but possible distinct sources of slow inhibition remain unknown. We identified a class of GABAergic interneurons, the neurogliaform cells, that, in contrast to other GABA-releasing cells, elicited combined GABA(A) and GABA(B) receptor-mediated responses with single action potentials and that predominantly targeted the dendritic spines of pyramidal neurons. Slow inhibition evoked by a distinct interneuron in spatially restricted postsynaptic compartments could locally and selectively modulate cortical excitability. PMID- 12649486 TI - Origin of the superflock of cichlid fishes from Lake Victoria, East Africa. AB - Lake Victoria harbors a unique species-rich flock of more than 500 endemic haplochromine cichlid fishes. The origin, age, and mechanism of diversification of this extraordinary radiation are still debated. Geological evidence suggests that the lake dried out completely about 14,700 years ago. On the basis of phylogenetic analyses of almost 300 DNA sequences of the mitochondrial control region of East African cichlids, we find that the Lake Victoria cichlid flock is derived from the geologically older Lake Kivu. We suggest that the two seeding lineages may have already been lake-adapted when they colonized Lake Victoria. A haplotype analysis further shows that the most recent desiccation of Lake Victoria did not lead to a complete extinction of its endemic cichlid fauna and that the major lineage diversification took place about 100,000 years ago. PMID- 12649487 TI - Gating the selectivity filter in ClC chloride channels. AB - ClC channels conduct chloride (Cl-) ions across cell membranes and thereby govern the electrical activity of muscle cells and certain neurons, the transport of fluid and electrolytes across epithelia, and the acidification of intracellular vesicles. The structural basis of ClC channel gating was studied. Crystal structures of wild-type and mutant Escherichia coli ClC channels bound to a monoclonal Fab fragment reveal three Cl- binding sites within the 15-angstrom neck of an hourglass-shaped pore. The Cl- binding site nearest the extracellular solution can be occupied either by a Cl- ion or by a glutamate carboxyl group. Mutations of this glutamate residue in Torpedo ray ClC channels alter gating in electrophysiological assays. These findings reveal a form of gating in which the glutamate carboxyl group closes the pore by mimicking a Cl- ion. PMID- 12649488 TI - Role of histone H3 lysine 27 methylation in X inactivation. AB - The Polycomb group (PcG) protein Eed is implicated in regulation of imprinted X chromosome inactivation in extraembryonic cells but not of random X inactivation in embryonic cells. The Drosophila homolog of the Eed-Ezh2 PcG protein complex achieves gene silencing through methylation of histone H3 on lysine 27 (H3-K27), which suggests a role for H3-K27 methylation in imprinted X inactivation. Here we demonstrate that transient recruitment of the Eed-Ezh2 complex to the inactive X chromosome (Xi) occurs during initiation of X inactivation in both extraembryonic and embryonic cells and is accompanied by H3-K27 methylation. Recruitment of the complex and methylation on the Xi depend on Xist RNA but are independent of its silencing function. Together, our results suggest a role for Eed-Ezh2-mediated H3 K27 methylation during initiation of both imprinted and random X inactivation and demonstrate that H3-K27 methylation is not sufficient for silencing of the Xi. PMID- 12649489 TI - Rube Goldberg goes (ribo)nuclear? Molecular switches and sensors made from RNA. AB - Switches and sensors play important roles in our everyday lives. The chemical properties of RNA make it amenable for use as a switch or sensor, both artificially and in nature. This review focuses on recent advances in artificial RNA switches and sensors. Researchers have been applying classical biochemical principles such as allostery in elegant ways that are influencing the development of biosensors and other applications. Particular attention is given here to allosteric ribozymes (aptazymes) that are regulated by small organic molecules, by proteins, or by oligonucleotides. Also discussed are ribozymes whose activities are controlled by various nonallosteric strategies. PMID- 12649490 TI - A twisted tRNA intermediate sets the threshold for decoding. AB - Putting together consistent cryo-EM structure, transient kinetic and mutant tRNA suppressor data, it appears that a deformed or waggling aminoacyl-tRNA is the critical transitional structure examined by the ribosome during decoding at the A site. The unusual conformation may be required for effective proofreading of the codon-anticodon complex. PMID- 12649491 TI - Recognition of acceptor-stem structure of tRNA(Asp) by Escherichia coli aspartyl tRNA synthetase. AB - Protein-RNA recognition between aminoacyl-tRNA synthetases and tRNA is highly specific and essential for cell viability. We investigated the structure-function relationships involved in the interaction of the Escherichia coli tRNA(Asp) acceptor stem with aspartyl-tRNA synthetase. The goal was to isolate functionally active mutants and interpret them in terms of the crystal structure of the synthetase-tRNA(Asp) complex. Mutants were derived from Saccharomyces cerevisiae tRNA(Asp), which is inactive with E. coli aspartyl-tRNA synthetase, allowing a genetic selection of active tRNAs in a tRNA(Asp) knockout strain of E. coli. The mutants were obtained by directed mutagenesis or library selections that targeted the acceptor stem of the yeast tRNA(Asp) gene. The mutants provide a rich source of tRNA(Asp) sequences, which show that the sequence of the acceptor stem can be extensively altered while allowing the tRNA to retain substantial aminoacylation and cell-growth functions. The predominance of tRNA backbone-mediated interactions observed between the synthetase and the acceptor stem of the tRNA in the crystal and the mutability of the acceptor stem suggest that many of the corresponding wild-type bases are replaceable by alternative sequences, so long as they preserve the initial backbone structure of the tRNA. Backbone interactions emerge as an important functional component of the tRNA-synthetase interaction. PMID- 12649492 TI - RNA aptamers to initiation factor 4A helicase hinder cap-dependent translation by blocking ATP hydrolysis. AB - The mammalian translation initiation factor 4A (eIF4A) is a prototype member of the DEAD-box RNA helicase family that couples ATPase activity to RNA binding and unwinding. In the crystal form, eIF4A has a distended "dumbbell" structure consisting of two domains, which probably undergo a conformational change, on binding ATP, to form a compact, functional structure via the juxtaposition of the two domains. Moreover, additional conformational changes between two domains may be involved in the ATPase and helicase activity of eIF4A. The molecular basis of these conformational changes, however, is not understood. Here, we generated RNA aptamers with high affinity for eIF4A by in vitro RNA selection-amplification. On binding, the RNAs inhibit ATP hydrolysis. One class of RNAs contains members that exhibit dissociation constant of 27 nM for eIF4A and severely inhibit cap dependent in vitro translation. The binding affinity was increased on Arg substitution in the conserved motif Ia of eIF4A, which probably improves a predicted arginine network to bind RNA substrates. Selected RNAs, however, failed to bind either domain of eIF4A that had been split at the linker site. These findings suggest that the selected RNAs interact cooperatively with both domains of eIF4A, either in the dumbbell or the compact form, and entrap it into a dead end conformation, probably by blocking the conformational change of eIF4A. The selected RNAs, therefore, represent a new class of specific inhibitors that are suitable for the analysis of eukaryotic initiation, and which pose a potential therapeutic against malignancies that are caused by aberrant translational control. PMID- 12649493 TI - SsrA-mediated trans-translation plays a role in mRNA quality control by facilitating degradation of truncated mRNAs. AB - An important unsolved question regarding the bacterial SsrA system is the fate of target mRNAs replaced by SsrA RNA during trans-translation. The aim of the present study is to address the potential role of SsrA system in mRNA quality control, focusing on truncated mRNAs that are expected to arise from 3'-to-5' exonucleolytic attack. We found that significant amounts of truncated mRNAs and polypeptides were produced from genes lacking a rho-independent terminator in SsrA-deficient cells. These truncated mRNAs, hence truncated polypeptides, were no longer observed in the presence of SsrA RNA. The data indicate that the SsrA system facilitates degradation of "nonstop" mRNAs by presumably removing the stalled ribosomes. Furthermore, analysis of affinity-purified proteins indicated that truncated polypeptides could be produced even from a gene with an intact rho independent terminator, although less efficiently, implying that C-terminally truncated proteins and 3'-truncated mRNA may be produced from virtually all protein-coding genes. We conclude that the SsrA system not only promotes the degradation of incomplete polypeptides but also minimizes the synthesis of incomplete polypeptides by facilitating the degradation of truncated mRNAs that are produced in cells. PMID- 12649494 TI - Antagonistic signals within the COX2 mRNA coding sequence control its translation in Saccharomyces cerevisiae mitochondria. AB - Translation of the mitochondrially coded COX2 mRNA within the organelle in yeast produces the precursor of Cox2p (pre-Cox2p), which is processed and assembled into cytochrome c oxidase. The mRNA sequence of the first 14 COX2 codons, specifying the pre-Cox2p leader peptide, was previously shown to contain a positively acting element required for translation of a mitochondrial reporter gene, ARG8(m), fused to the 91st codon of COX2. Here we show that three relatively short sequences within the COX2 mRNA coding sequence, or structures they form in vivo, inhibit translation of the reporter in the absence of the positive element. One negative element was localized within codons 15 to 25 and shown to function at the level of the mRNA sequence, whereas two others are within predicted stem-loop structures formed by codons 22-44 and by codons 46-74. All three of these inhibitory elements are antagonized in a sequence-specific manner by reintroduction of the upstream positive-acting sequence. These interactions appear to be independent of 5'- and 3'-untranslated leader sequences, as they are also observed when the same reporter constructs are expressed from the COX3 locus. Overexpression of MRS2, which encodes a mitochondrial magnesium carrier, partially suppresses translational inhibition by each isolated negatively acting element, but does not suppress them in combination. We hypothesize that interplay among these signals during translation in vivo may ensure proper timing of pre-Cox2p synthesis and assembly into cytochrome c oxidase. PMID- 12649495 TI - Efficient transcription of the EBER2 gene depends on the structural integrity of the RNA. AB - A 3'-truncated EBER2 RNA gene, although containing all previously identified promoter elements, revealed drastically reduced transcription rates in vitro and in vivo when fused to a heterologous terminator sequence. Inactivations were also observed with double point mutations affecting 5'- or 3'-end sequences of the EBER2 gene. However, wild-type activity of these mutants could be restored by compensatory mutations of the opposite strand of the EBER2 RNA sequence. A similar rescue was achieved with the 3'-truncated EBER2 gene, if the heterologous terminator was adapted for complementarity to the initiator element of the construct. Yet, double-strandedness alone of the RNA ends was not sufficient for high transcriptional activity of these gene constructs. Rather, the use of a nonrefoldable spacer, separating the 5'- and 3'-stem-loop structures, demonstrated that spatial proximity of the ends of EBER2 RNA was required. Furthermore, decay kinetics of wild-type and mutant RNA synthesized in vitro indicated that the effects observed could not be explained by altered transcript stability. Finally, single-round transcription confirmed that the reduced expression of mutant genes was not caused by decreased primary initiation reactions. In addition, differential sarcosyl concentrations demonstrated that the rate of reinitiation clearly was affected with the mutant EBER2 genes. Together, these results indicate that the secondary structure of this viral RNA represents a major determinant for efficient transcription of the EBER2 gene by host cell RNA polymerase III. PMID- 12649496 TI - Antagonistic regulation of alpha-actinin alternative splicing by CELF proteins and polypyrimidine tract binding protein. AB - The alpha-actinin gene has a pair of alternatively spliced exons. The smooth muscle (SM) exon is repressed in most cell types by polypyrimidine tract binding protein (PTB). CELF (CUG-BP and ETR3-like factors) family proteins, splicing regulators whose activities are altered in myotonic dystrophy, were found to coordinately regulate selection of the two alpha-actinin exons. CUG-BP and ETR3 activated the SM exon, and along with CELF4 they were also able to repress splicing of the NM (nonmuscle) exon both in vivo and in vitro. Activation of SM exon splicing was associated with displacement of PTB from the polypyrimidine tract by binding of CUG-BP at adjacent sites. Our data provides direct evidence for the activity of CELF proteins as both activators and repressors of splicing within a single-model system of alternative splicing, and suggests a model whereby alpha-actinin alternative splicing is regulated by synergistic and antagonistic interactions between members of the CELF and PTB families. PMID- 12649497 TI - RBP16 is a multifunctional gene regulatory protein involved in editing and stabilization of specific mitochondrial mRNAs in Trypanosoma brucei. AB - RBP16 is a Trypanosoma brucei mitochondrial RNA-binding protein that associates with guide RNAs (gRNAs), mRNAs, and ribosomal RNAs. Based on its inclusion in the multifunctional Y-box protein family and its ability to bind multiple RNA classes, we hypothesized that RBP16 plays a role in diverse aspects of mitochondrial gene regulation. To gain insight into RBP16 function, we generated cells expressing less than 10% of wild-type RBP16 levels by tetracycline regulated RNA interference (RNAi). Poisoned primer extension analyses revealed that edited, but not unedited, CYb mRNA is reduced by approximately 98% in tetracycline-induced RBP16 RNAi cells, suggesting that RBP16 is critical for CYb RNA editing. The down-regulation of CYb editing in RBP16 RNAi transfectants apparently entails a defect in gRNA utilization, as gCYb[560] abundance is similar in uninduced and induced cells. We observed a surprising degree of specificity regarding the ability of RBP16 to modulate editing, as editing of mRNAs other than CYb is not significantly affected upon RBP16 disruption. However, the abundance of the never edited mitochondrial RNAs COI and ND4 is reduced by 70%-80% in RBP16 RNAi transfectants, indicating an additional role for RBP16 in the stabilization of these mRNAs. Analysis of RNAs bound to RBP16 immunoprecipitated from wild-type cells reveals that RBP16 is associated with multiple gRNA sequence classes in vivo, including those whose abundance and usage appear unaffected by RBP16 disruption. Overall, our results indicate that RBP16 is an accessory factor that regulates the editing and stability of specific populations of mitochondrial mRNAs. PMID- 12649498 TI - Sequence and structural requirements for optimal guide RNA-directed insertional editing within Leishmania tarentolae. AB - The coding sequence of several mitochondrial mRNAs of the trypanosomatid family of protozoa is created by the guide RNA-directed insertion and deletion of uridylates (Us). Selection-amplification was used to explore the sequence and structure of the guide RNA and mRNA required for efficient insertional editing within a mitochondrial extract prepared from Leishmania tarentolae. This study identifies several novel features of the editing reaction in addition to several that are consistent with the previous mutagenesis and phylogenetic analysis of the reaction in Trypanosoma brucei, a distantly related trypanosomatid. Specifically, there is a strong bias against cytidines 5' of the editing sites and guanosines immediately 3' of guiding nucleotides. U insertions are directed both 5' and 3' of a genomically encoded U, which was previously assumed not to occur. Base pairing immediately flanking an editing site can significantly stimulate the editing reaction and affect the reaction fidelity but is not essential. Likewise, single-stranded RNA in the region upstream of the editing site, not necessarily immediately adjacent, can facilitate editing but is also not essential. The editing of an RNA containing many of the optimal features is linear with increasing quantities of extract permitting specific activity measurements to be made that are not possible with previously described T. brucei and L. tarentolae assays. The reaction catalyzed by the L. tarentolae extract can be highly accurate, which does not support a proposed model for editing that was based largely on the inaccuracy of an earlier in vitro reaction. PMID- 12649499 TI - Identification of novel components of Trypanosoma brucei editosomes. AB - The editosome is a multiprotein complex that catalyzes the insertion and deletion of uridylates that occurs during RNA editing in trypanosomatids. We report the identification of nine novel editosome proteins in Trypanosoma brucei. They were identified by mass spectrometric analysis of functional editosomes that were purified by serial ion exchange/gel permeation chromatography, immunoaffinity chromatography specific to the TbMP63 editosome protein, or tandem affinity purification based on a tagged RNA editing ligase. The newly identified proteins have ribonuclease and/or RNA binding motifs suggesting nuclease function for at least some of these. Five of the proteins are interrelated, as are two others, and one is related to four previously identified editosome proteins. The implications of these findings are discussed. PMID- 12649505 TI - Clinical relevance of atrial fibrillation/flutter, stroke, pacemaker implant, and heart failure in Emery-Dreifuss muscular dystrophy: a long-term longitudinal study. AB - BACKGROUND AND PURPOSE: Emery-Dreifuss muscular dystrophy (EDMD) is a rare inherited disorder associated with cardiac involvement. We investigated the spectrum and relevance of the cardiac manifestations of EDMD, focusing on bradyarrhythmias and tachyarrhythmias (including atrial fibrillation/flutter), embolic stroke, and heart failure. METHODS AND RESULTS: Eighteen patients (age 42.8+/-19.6 years) with genetically confirmed X-linked (n=10, including 3 carriers) or autosomal dominant (n=8) EDMD were followed for a period ranging from 1 to 30 years in a research center for neuromuscular diseases and in a university cardiological department. Pacemakers were required by 10 of 18 (56%) patients for bradyarrhythmia, and related complications occurred in 3 of 10 (30%) cases. Atrial fibrillation/flutter developed in 11 of 18 (61%) patients, with atrial standstill subsequently occurring in 5 of 11 (45%) cases and embolic stroke (most often disabling) in 4 of 11 (36%). Heart failure requiring transplantation occurred in 1 of 18 (6%) patients, and asymptomatic left ventricular dysfunction in a further 3 (17%). No relationship was evident between neuromuscular impairment and cardiac involvement. CONCLUSIONS: Both X-linked and autosomal dominant EDMD patients risk not only bradyarrhythmia (requiring pacemaker implant) but also atrial fibrillation/flutter, which often anticipates atrial standstill and can cause disabling embolic stroke at a relatively young age. Antithromboembolic prophylaxis has to be recommended in EDMD patients with atrial fibrillation/flutter or atrial standstill. With careful monitoring, survival after pacemaker implant may be long. Heart failure, which seems to occur only in a minority of patients, may be severe. PMID- 12649500 TI - Lentivirus-delivered stable gene silencing by RNAi in primary cells. AB - Genome-wide genetic approaches have proven useful for examining pathways of biological significance in model organisms such as Saccharomyces cerevisiae, Drosophila melanogastor, and Caenorhabditis elegans, but similar techniques have proven difficult to apply to mammalian systems. Although manipulation of the murine genome has led to identification of genes and their function, this approach is laborious, expensive, and often leads to lethal phenotypes. RNA interference (RNAi) is an evolutionarily conserved process of gene silencing that has become a powerful tool for investigating gene function by reverse genetics. Here we describe the delivery of cassettes expressing hairpin RNA targeting green fluorescent protein (GFP) using Moloney leukemia virus-based and lentivirus-based retroviral vectors. Both transformed cell lines and primary dendritic cells, normally refractory to transfection-based gene transfer, demonstrated stable silencing of targeted genes, including the tumor suppressor gene TP53 in normal human fibroblasts. This report demonstrates that both Moloney leukemia virus and lentivirus vector-mediated expression of RNAi can achieve effective, stable gene silencing in diverse biological systems and will assist in elucidating gene functions in numerous cell types including primary cells. PMID- 12649506 TI - Bedside monitoring of circulating blood volume after subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: Maintenance of an adequate intravascular volume is important in the management of patients with subarachnoid hemorrhage (SAH). The purpose of this study was to investigate the circulating blood volume (CBV) after SAH with the use of indocyanine green pulse spectrophotometry. METHODS: CBV and plasma hormones related to stress and fluid regulation were measured 4 times: day 2 to 3, day 4 to 5, day 7 to 8, and day 14 in 50 consecutive patients with SAH surgically treated within 48 hours. RESULTS: The mean value of CBV was 64 mL/kg on day 2 to 3, which gradually increased to 69 mL/kg on day 4 to 5, 71 mL/kg on day 7 to 8, and 70 mL/kg on day 14 (P=0.005) (control, 72 mL/kg). The clinical grades and plasma corticotropin levels were higher in patients with <60 mL/kg of CBV on day 2 to 3 (P<0.05 for both). There were no significant differences in other physiological and laboratory parameters such as time for surgery, estimated blood loss, levels of plasma noradrenaline, brain natriuretic peptide, serum sodium, and hematocrit. When CBV was decreased >10% of the former level, there were decreases in hematocrit (P<0.05), serum sodium (P<0.01), and serum albumin (P<0.05) and an increase in urinary sodium (P<0.05). CONCLUSIONS: A significant reduction of CBV, especially in patients with poor clinical grades, was noted after SAH and early surgery, which could not be detected by routine examinations. Anemia, central salt wasting, and hypoalbuminemia may be related to a decrease in CBV from the former level. Indocyanine green pulse spectrophotometry may be a powerful tool for the management of patients with SAH. PMID- 12649507 TI - Increased expression of neuronal apolipoprotein E in human brain with cerebral infarction. AB - BACKGROUND AND PURPOSE: Cellular origin of apolipoprotein E (ApoE) in the human brain and its roles in physiological and pathological conditions remain to be clarified. METHODS: Immunolocalization of ApoE was investigated in a series of autopsied human brains with or without infarction. ApoE expression was also estimated on immunoblot on protein extracts from autopsied brains and a cultured neuroblastoma cell line of human origin (GOTO) subjected to an oxidative stress induced by exposure to hydrogen peroxide (0.2 mmol/L). RESULTS: In addition to astrocytes and microglia, neurons and degenerated axons in and around the ischemic foci contained ApoE-like immunoreactivity, which was more intense in recent ischemic foci. Immunoblot demonstrated an increase in expression of ApoE in brain extracts from ischemic lesion, and this increase was also pronounced in the cultured neuroblastoma cell line after the stress. CONCLUSIONS: Accumulation of ApoE in neurons in and around ischemic foci of the human brain is related to an increase in ApoE synthesis in neurons, as seen in cultured neuronal cells after oxidative stress. Intrinsic regenerative activity of neuron in reaction to external insults may be related to this increase in ApoE of neuronal origin. PMID- 12649508 TI - Opening of mitochondrial ATP-sensitive potassium channels is a trigger of 3 nitropropionic acid-induced tolerance to transient focal cerebral ischemia in rats. AB - BACKGROUND AND PURPOSE: The role of mitochondrial ATP-sensitive potassium channels (mitoK(ATP)) in ischemic tolerance has been well documented in heart, but little work has been done in brain. To investigate the involvement of mitoK(ATP) activation in chemical preconditioning in brain, we examined the effect of 5-hydroxydecanoate (5-HD), a selective mitoK(ATP) blocker, on neurotoxin 3-nitropropionic acid (3-NPA)-induced ischemic tolerance to transient focal cerebral ischemia in rats. METHODS: Male Wistar rats were administrated 3 NPA (20 mg/kg IP; n=16) or vehicle (saline; n=16) 3 days before temporary occlusion (120 minutes) of the middle cerebral artery; 5-HD (40 mg/kg IP; n=16) was injected 20 minutes before 3-NPA administration. Infarct volumes were measured 4 days after reperfusion. To directly investigate whether chemical preconditioning activates mitoK(ATP), we tested the effect of prior incubation with 1 mmol/L 5-HD on 300 micromol/L 3-NPA-induced alterations of mitochondrial membrane potential (Delta(Psi)m) in cultured neurons and astrocytes using the fluorescent dye tetramethylrhodamine ethyl ester. RESULTS: Treatment with 3-NPA exhibited a 16% reduction (P<0.05) and 23% reduction in infarct volume (P<0.01) for total brain and cortex, respectively. Pretreatment with 5-HD completely abolished the neuroprotective effect of chemical preconditioning. In cultured cells, 3-NPA resulted in mitochondrial depolarization. This change of Delta(Psi)m was completely blocked by 5-HD pretreatment. CONCLUSIONS: These results strongly suggest that opening of mitoK(ATP) plays a key role as the trigger in the development of 3-NPA-induced ischemic tolerance in brain. PMID- 12649509 TI - Constraint-induced movement therapy and rehabilitation exercises lessen motor deficits and volume of brain injury after striatal hemorrhagic stroke in rats. AB - BACKGROUND AND PURPOSE: Constraint-induced movement therapy (CIMT) promotes motor recovery after occlusive stroke in humans, but its efficacy after intracerebral hemorrhage (ICH) has not been investigated clinically or in the laboratory. In this study we tested whether CIMT and a rehabilitation exercise program would lessen motor deficits after ICH in rats. METHODS: Rats were subjected to striatal ICH (via infusion of collagenase) or sham stroke. Seven days later, treatment began with CIMT (8 h/d of ipsilateral forelimb restraint), rehabilitation exercises (eg, reaching, walking; 1 h/d), or both for 7 days. Some rats were not treated. Motor deficits were assessed up to the 60-day survival time, after which the volume of tissue lost was determined. RESULTS: Untreated ICH rats made more limb slips traversing a horizontal ladder and showed an asymmetry toward less use of the contralateral paw in the cylinder test of limb use asymmetry (day 28). These rats were also significantly less successful in the Montoya staircase test (days 55 to 59) of skilled reaching. Neither therapy alone provided much benefit. However, the combination of daily exercises and CIMT substantially and persistently improved recovery. Unexpectedly, this group had a statistically smaller volume of tissue lost than untreated ICH rats. CONCLUSIONS: The combination of focused rehabilitation exercises and CIMT effectively promotes functional recovery after ICH, while either therapy alone is less effective. This therapy may work in part by reducing the volume of tissue lost, likely through reducing atrophy while promoting remodeling. PMID- 12649510 TI - Stereotactic treatment of intracerebral hematoma by means of a plasminogen activator: a multicenter randomized controlled trial (SICHPA). AB - BACKGROUND AND PURPOSE: Treatment of intracerebral hematoma (ICH) is controversial. An advantage of neurosurgical intervention over conservative treatment of ICH has not been established. Recent reports suggest a favorable effect of stereotactic blood clot removal after liquefaction by means of a plasminogen activator. The SICHPA trial was aimed at investigating the efficacy of this treatment. METHODS: A stereotactically placed catheter was used to instill urokinase to liquefy and drain the ICH in 6-hour intervals over 48 hours. From 1996 to 1999, 13 centers entered 71 patients into the study. Patients were randomized into a surgical group (n=36) and a nonsurgical group (n=35). Admission criteria were the following: age >45 years, spontaneous supratentorial ICH, Glasgow Eye Motor score ranging from 2 to 10, ICH volume >10 cm3, and treatment within 72 hours. The primary end point was death at 6 months. As secondary end points, ICH volume reduction and overall outcome measured by the modified Rankin scale were chosen. The trial was prematurely stopped as a result of slow patient accrual. RESULTS: Seventy patients were analyzed. Overall mortality at day 180 after stroke was 57%; this included 20 of 36 patients (56%) in the surgical group and 20 of 34 patients (59%) in the nonsurgical group. A significant ICH volume reduction was achieved by the intervention (10% to 20%, P<0.05). Logistic regression analysis indicated the possibility of efficacy for surgical treatment (odds ratio, 0.23; 95% confidence interval, 0.05 to 1.20; P=0.08). The odds ratio of mortality combined with modified Rankin scale score 5 at 180 days was also not statistically significant (odds ratio, 0.52; 95% confidence interval, 1.2 to 2.3; P=0.38). CONCLUSIONS: Stereotactic aspiration can be performed safely and in a relatively uniform manner; it leads to a modest reduction of 18 mL of hematoma reduction over 7 days when compared with control, which has a 7-mL reduction, and therefore may improve prognosis. PMID- 12649511 TI - Emergency calls in acute stroke. AB - BACKGROUND AND PURPOSE: In the last 10 years, stroke has become a medical emergency. Subsequently, early recognition of stroke symptoms and rapid activation of the medical system are essential. We sought to investigate what witnesses or victims of an acute stroke syndrome recognize and report in the actual situation. METHODS: We analyzed the recordings of all patients admitted to our stroke unit via the Emergency Medical System (EMS) dispatch center in Nuremberg within 1 year. With a structured evaluation form, the calls were screened for symptoms reported and for any diagnosis or other facts mentioned spontaneously or in response to a question by the dispatcher. We also evaluated data about EMS response and patient condition on admission. RESULTS: Of 482 patients treated in our stroke unit, 141 calls were evaluated. Main symptoms reported included speech problems (25.5%), motor deficits (21.9%), and disturbances of consciousness (14.8%). In many cases, a fall (21.2%) was presented as the main problem. Sensory deficits (7.8%) and vertigo (5.6%) were rarely mentioned. In 28 calls (19.8%), stroke was mentioned as a possible cause of the acute health problems. The dispatcher suspected a stroke in 51.7% of all cases. CONCLUSIONS: This is one of the first studies to investigate emergency calls in acute stroke. We found that motor deficits and speech problems were the most dramatic symptoms that led to activation of the EMS. Other symptoms were less frequently reported, or atypical descriptions were given. Educational efforts are needed to improve recognition of atypical stroke symptoms by stroke victims and EMS professionals. PMID- 12649512 TI - Increasing stroke incidence and decreasing case fatality, 1989-1998: a study from the stroke register in Malmo, Sweden. AB - BACKGROUND AND PURPOSE: Although the incidence of and mortality from myocardial infarction (MI) have declined in most industrialized countries, incidence studies of stroke have shown less consistent trends. This study examines time trends in the incidence of stroke and case fatality rates in relation to socioeconomic circumstances and history of MI. METHODS: Stroke incidence in the city of Malmo, Sweden (250,000 citizens), has been continuously monitored since 1989. All patients 50 to 79 years of age who experienced a first-ever stroke between 1989 and 1998 were included. RESULTS: We included 3621 patients (1969 men, 1652 women). The age-standardized incidence was 647 per 100,000 persons-years for men and 400 per 100,000 persons-years for women. The annual increase-3.1% in men (P<0.05) and 2.9% in women (P<0.05)-was more pronounced in the younger age groups and was lowest in areas with poor socioeconomic circumstances. We found that 13% of the men and 6% of the women had a history of MI; this proportion was stable over the study period. The average case fatality rates in 1989 to 1998 were 10% for men and 9.3% for women. Rates remained stable for men but declined significantly in women (odds ratio per year, 0.895; 95% confidence interval, 0.84 to 0.95). CONCLUSIONS: In this urban population, stroke incidence increased between 1989 and 1998. The rate of increase tended to be lower in residential areas with least favorable socioeconomic circumstances. The proportion with a history of MI remained unchanged during the study period. There has been significant improvement in the case fatality rate in women but not in men. PMID- 12649513 TI - Veterans Administration Acute Stroke (VASt) Study: lack of race/ethnic-based differences in utilization of stroke-related procedures or services. AB - BACKGROUND AND PURPOSE: Race/ethnic-based disparities in the utilization of health-related services have been reported. Data collected as part of the Veterans Administration Acute Stroke Study (VASt) were analyzed to determine whether similar differences were present in patients admitted to Veterans Administration (VA) hospitals with acute ischemic stroke. METHODS: VASt prospectively identified stroke patients admitted to 9 geographically separated VA hospitals between April 1995 and March 1997. Demographic characteristics and all inpatient diagnostic tests/procedures were recorded. Frequencies were compared with chi2 tests. RESULTS: Of 1073 enrolled patients, 775 (white, n=520; nonwhite, n=255, including 226 blacks and 28 Hispanic-Americans) with ischemic stroke were admitted from home. Mean ages (71.0+/-0.6 versus 71.9+/-0.4 years; P=0.25) and Trial of ORG 10172 in Acute Stroke Treatment (TOAST) stroke types (atherothrombotic, 12.9% versus 13.3%; cardioembolic, 16.5% versus 18.0%; lacunar, 26.4% versus 27.1%; other, 1.4% versus 2.0%; unclassified, 42.9% versus 39.6%; P=0.89) for whites versus nonwhites were similar. There were no race/ethnic-based differences in the utilization of brain CT (91.0% versus 92.2%; P=0.58), MRI (36.2% versus 41.6%; P=0.14), transthoracic (52.5% versus 53.7%; P=0.75) or transesophageal echocardiography (10.2% versus 10.6%; P=0.86), 24-hour ECG (3.3% versus 1.6%; P=0.17), carotid ultrasound (64.0% versus 62.0%; P=0.57), carotid endarterectomy (1.5% versus 0.8%; P=0.38), rehabilitation evaluations (71.0% versus 76.5%; P=0.11), speech therapy (9.6% versus 12.6%; P=0.21), recreational therapy (3.1% versus 2.0%; P=0.37), or occupational therapy (16.0% versus 19.6%; P=0.20) for whites versus nonwhites, respectively. Angiography was performed less frequently (3.1% versus 8.5%; P=0.01) and ECG more frequently (81.6% versus 73.5%; P=0.01) in nonwhites. The proportions of patients discharged functionally independent were also similar (52% of whites and 50% of nonwhites had discharge Rankin Scale scores of 0, 1, or 2; P=0.63). CONCLUSIONS: Aside from cerebral angiography and ECG, there were no race/ethnic-based disparities in the utilization of a variety of stroke-related procedures and services. The difference in the use of angiography is unlikely to be related to a difference in screening for carotid endarterectomy because there was no difference in the frequency of carotid ultrasonography. The reason ECG was obtained more frequently in nonwhites is uncertain. PMID- 12649514 TI - Recommendations for the relationship between sponsors and investigators in the design and conduct of clinical stroke trials. AB - BACKGROUND: As increasing numbers of clinical trials are being conducted worldwide, there has been concern about the relationship between the sponsors and investigators. Both parties need benchmarks or recommendations to act as a reference point when trials are initiated and conducted. These recommendations are designed to fulfill this role. SUMMARY OF COMMENT: A series of meetings of the International Trial Subcommittee of the International Stroke Liaison Committee, American Stroke Association, were conducted to review a series of draft recommendations that were then modified on the basis of the experience of the committee in the conduct of clinical trials. Consensus was reached on all points of the final document. The recommendations represent the opinions of the authors and do not necessarily reflect the views of the American Stroke Association. The document contains sensible recommendations concerning required sponsor company qualities, trial management structure, protocol development, trial conduct, data management, blinding, data analyses, publication, remuneration, and conflict of interest issues. CONCLUSIONS: The recommendations should provide a simple and practical benchmark for both investigators and sponsors in the conduct of clinical trials. Although designed for trials involving therapies for stroke, the framework allows generalization to other disciplines. PMID- 12649515 TI - Comparison of triflusal and aspirin for prevention of vascular events in patients after cerebral infarction: the TACIP Study: a randomized, double-blind, multicenter trial. AB - BACKGROUND AND PURPOSE: The efficacy of the antiplatelet agent triflusal for prevention of vascular events after stroke has been reported in a pilot study. However, there is a need to confirm those results in a larger study. METHODS: We performed a randomized, double-blind, multicenter study to test the efficacy of triflusal (600 mg/d) versus aspirin (325 mg/d) for prevention of vascular events in patients with stroke or transient ischemic attack (Triflusal versus Aspirin in Cerebral Infarction Prevention [TACIP]). We assessed a combined end point (incidence of nonfatal ischemic stroke, nonfatal acute myocardial infarction, or vascular death) as well as the incidence of these events separately and the incidence of major hemorrhage. RESULTS: Of 2113 patients, 1058 received triflusal and 1055 aspirin. The mean follow-up period was 30.1 months. The incidence of combined end point (13.1% for triflusal, 12.4% for aspirin) as well the survival analysis (hazard ratio [HR] for triflusal versus aspirin, 1.09; 95% CI, 0.85 to 1.38) showed no differences between groups. The incidence of nonfatal stroke (HR, 1.09; 95% CI, 0.82 to 1.44), nonfatal acute myocardial infarction (HR, 0.95; 95% CI, 0.46 to 1.98,) and vascular death (HR, 1.22; 95% CI, 0.75 to 1.96) was also similar. A significantly higher incidence of major hemorrhages in the aspirin group was recorded (HR, 0.48; 95% CI, 0.28 to 0.82). The overall incidence of hemorrhage was significantly lower in the triflusal group (16.7% versus 25.2%) (odds ratio, 0.76; 95% CI, 0.67 to 0.86; P<0.001). CONCLUSIONS: This study failed to show significantly superior efficacy of triflusal over aspirin in the long term prevention of vascular events after stroke, but triflusal was associated with a significantly lower rate of hemorrhagic complications. PMID- 12649516 TI - Feasibility of radioactive embolization of intracranial aneurysms using 32P implanted coils. AB - BACKGROUND AND PURPOSE: Beta radiation can prevent recanalization after embolization. Our goal was to assess the feasibility of endovascular treatment of intracranial aneurysms using coils of a predetermined activity of 32P per centimeter. METHODS: We studied the total length of coils deployed into 357 intracranial aneurysms. Aneurysmal volumes were estimated using 3 mathematical models. We simulated that coils were implanted with 0.26 microCi/cm of 32P, calculated resulting volumetric activities, and compared them with "effective" levels derived from experimental data and "safe" levels prescribed for the clinical use of 32P in cystic craniopharyngiomas. RESULTS: Effective activities would have been reached in 92% to 98% of lesions had the coils been radioactive at the time of treatment. CONCLUSIONS: Radioactive coil embolization of aneurysms is feasible in most patients. PMID- 12649517 TI - Thrombin-activable fibrinolysis inhibitor levels in the acute phase of ischemic stroke. AB - BACKGROUND AND PURPOSE: Thrombin-activable fibrinolysis inhibitor (TAFI) is a recently identified fibrinolysis inhibitor in plasma. The purpose of this work was to study TAFI levels in the acute phase of ischemic stroke and their relationship with stroke evolution. METHODS: In 30 consecutive ischemic stroke patients, TAFI plasma levels were measured by means of enzyme-linked immunosorbent assay (percentage of the pooled reference kit expressed as mean+/ SD) and compared with the values obtained in 30 healthy control subjects. All samples were drawn within the first 24 hours after symptom onset (mean, 4.6 hours) and before any treatment was started. RESULTS: TAFI plasma concentration was significantly higher (P<0.001) in stroke patients (158.4+/-53.2%) than in healthy control subjects (105.6+/-30.2%). The highest TAFI levels were found in cases of neurological deterioration (worsening, 198.1+/-63.0%; stability, 130.5+/ 39.3%; improvement, 173.9+/-52.0%; P=0.057). CONCLUSIONS: High levels of TAFI are found in the acute phase of ischemic stroke. PMID- 12649518 TI - Prognostication of recovery in patients with acute ischemic stroke through the use of brain SPECT with Technetium-99m--labeled metronidazole. AB - BACKGROUND AND PURPOSE: We hypothesized that technetium-99m-ethylene dicysteine metronidazole (99mTc-EC-MN) localizes to brain tissue that is hypoxic but viable. This study prospectively evaluated the relationship between neurological outcome and uptake of 99mTc-EC-MN in peri-infarcted regions of the brain. METHODS: Eight patients with acute ischemic stroke in the territory of the left middle cerebral artery underwent 99mTc-EC-MN and 99mTc-ethyl cysteinate dimer (ECD) brain SPECTs on the same day during the subacute stage (10.3+/-2.5 days). The infarct volumes from 99mTc-ECD images (IV(ECD)), infarct volumes from diffusion-weighted MRI images (IV(DW)), and hypoxic volume (HV) from 99mTc-EC-MN images were calculated. The net infarct volume (NIV(ECD)) was defined as IV(ECD) minus HV. The National Institutes of Health Stroke Scale scores were measured on admission and days 1, 3, 7, and 30. RESULTS: IV(ECD) was greater than IV(DW). The lesion-to-normal count-density ratios of 99mTc-EC-MN ranged from 1.80 to 5.96. HV was 60.2+/-65.2 cm3, and the mean percent HV was 24.5+/-28.1% of IV(ECD). NIV(ECD) was 162.6+/ 133.4 cm3 and was significantly smaller than IV(ECD). NIV(ECD) was significantly correlated with National Institutes of Health Stroke Scale score at 1 month and was a significant predictor of neurological deficit at 1 month. CONCLUSIONS: 99mTc-EC-MN brain SPECT can detect hypoxic tissue after acute ischemic stroke and, in combination with 99mTc-ECD brain SPECT, is useful in predicting neurological outcome in ischemic stroke patients. PMID- 12649519 TI - Absence of linkage of familial intracranial aneurysms to 7q11 in highly aggregated Japanese families. AB - BACKGROUND AND PURPOSE: We sought to test the linkage of familial intracranial aneurysms (FIAs) to the ELN (elastin) locus in chromosome 7q11 reported previously. METHODS: Intracranial aneurysm (IA) probands were searched from patient records or neurosurgeons' recalls in collaborating hospitals. Members of the participating probands' families who had unknown affection status were screened by MR angiography and diagnosed by digital subtraction angiography. Inclusion criteria of families for genetic analyses were as follows: at least 3 alive affected members or 2 alive affected members with at least 1 unaffected member (>or=60 years). Linkage to the ELN locus was tested with the use of GENEHUNTER by parametric and nonparametric methods. To exclude false-negatives in the linkage analysis, the lowest 5% limits of logarithms of the odds (LOD) and nonparametric LOD (NPL) scores for individual families and for the total set of families were simulated on assumption that the ELN locus is linked to FIAs. RESULTS: Questionnaires were sent to 885 patients, and 563 responded. Seventy nine probands were positive for family history. One hundred thirty-four family members of unknown affection status were screened. A total of 14 families with 64 members met the criteria. Linkage to the ELN locus was discarded in 11 families and was inconclusive for 3 families. The total LOD and total NPL scores for 14 families were -8.04 and -0.643, respectively. Our conclusion did not change even when the values of penetrance were changed or only affected members were analyzed. CONCLUSIONS: The majority of aggregated IA Japanese families may not have a genetic linkage to chromosome 7q11. PMID- 12649520 TI - Anticoagulation for cerebral sinus thrombosis. PMID- 12649521 TI - Intensity of aphasia therapy, impact on recovery. AB - BACKGROUND: It has been speculated that the conflicting results demonstrated across poststroke aphasia therapy studies might be related to differences in intensity of therapy provided across studies. The aim of this study is to investigate the relationship between intensity of aphasia therapy and aphasia recovery. METHODS: A MEDLINE literature search was conducted to retrieve clinical trials investigating aphasia therapy after stroke. Changes in mean scores from each study were recorded. Intensity of therapy was recorded in terms of length of therapy, hours of therapy provided per week, and total hours of therapy provided. Pearson correlation was used to assess the relationship between changes in mean scores of outcome measures and intensity of therapy. RESULTS: Studies that demonstrated a significant treatment effect provided 8.8 hours of therapy per week for 11.2 weeks versus the negative studies that only provided approximately 2 hours per week for 22.9 weeks. On average, positive studies provided a total of 98.4 hours of therapy, whereas negative studies provided 43.6 hours of therapy. Total length of therapy time was found to be inversely correlated with hours of therapy provided per week (P=0.003) and total hours of therapy provided (P=0.001). Total length of therapy was significantly inversely correlated with mean change in Porch Index of Communicative Abilities (PICA) scores (P=0.0001). The number of hours of therapy provided in a week was significantly correlated to greater improvement on the PICA (P=0.001) and the Token Test (P=0.027). Total number of hours of therapy was significantly correlated with greater improvement on the PICA (P<0.001) and the Token Test (P<0.001). CONCLUSIONS: Intense therapy over a short amount of time can improve outcomes of speech and language therapy for stroke patients with aphasia. PMID- 12649522 TI - Abnormal expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in brain arteriovenous malformations. AB - BACKGROUND AND PURPOSE: Excessive degradation of the vascular matrix by matrix metalloproteinases (MMPs) can lead to structural instability of vessels. In this study we examined the expression of MMPs and tissue inhibitors of metalloproteinases (TIMPs) in brain arteriovenous malformations (BAVMs). METHODS: We performed gelatin zymography for MMPs and Western blot for MMP-9, MMP-2, TIMP 1, TIMP-2, TIMP-3, and TIMP-4. MMP-9 expression was localized by immunohistochemistry. RESULTS: We analyzed 37 BAVM specimens and 9 control brain specimens from epilepsy surgery. Thirty-two BAVM patients had embolization treatment before resection. Eighteen BAVM patients had a history of hemorrhage from BAVMs. Neither MMP-2 nor TIMP-2 was detected in BAVMs or control brain specimens. Compared with control brain samples, BAVM samples had higher levels of total MMP-9, active MMP-9, pro-MMP-9, TIMP-1, and TIMP-3. TIMP-4 levels were higher in the control brain than in BAVM specimens. MMP-9 was localized to the endothelial cell/peri-endothelial cell layer and infiltrating neutrophils of BAVMs. BAVMs with venous stenosis >or=50% had higher expression of MMP-9 than BAVMs with venous stenosis <50%. There was no apparent association between total MMP-9, pro-MMP-9, or active MMP-9 levels and (1) feeding artery pressure, (2) pattern of draining vein (exclusively deep venous drainage versus any superficial drainage), and (3) BAVM size. CONCLUSIONS: We found increased levels of MMP-9 and TIMPs in BAVMs. Abnormal balance of MMP-9 and TIMPs may contribute to vascular instability of BAVMs. PMID- 12649523 TI - William Feinberg lecture 2002: emotions, mood, and behavior after stroke. AB - While emotional outcome is a critical factor influencing early evolution and late prognosis after stroke, few relevant studies have been performed on this subject. However, mood changes, modified judgment, and emotional reactions may also dramatically alter recruitment into clinical trials; for instance, up to one third of patients with acute stroke may have altered time perception, inappropriate self-evaluation of their condition, and attentional or memory dysfunction, with a subsequent increase in referral-to-hospital delays. In addition, the value of the "informed" consenting process may be questionable in the setting of urgent randomization into an acute stroke clinical trial. Data from ongoing studies suggest that behavior and emotional reactions in acute stroke patients may be classified into a few broad categories, with considerable overlap. Correlations between mood changes and the type, location, and severity of stroke may provide useful information for improving patient management, including the prediction of functional evolution and late prognosis. While depressive reactions have been widely studied in the recovery-rehabilitation phase after stroke, significant depression is uncommon shortly after stroke. On the other hand, related, though different, emotional behavioral changes may be more frequent; these have often been confused with depression and include catastrophic reaction, emotionalism, and athymhormia. Late depression is the most common mood alteration during the first year after stroke and has specific characteristics that differentiate it from classic endogenous and reactive depression, thus emphasizing the critical role of brain damage in the pathogenesis of poststroke depression. Early recognition and management of mood disorders after stroke are critical for the functional improvement of individual patients. However, little is known about specific indications for different antidepressant drugs in poststroke depression and related disorders. Ongoing research has identified a "new" emotional-behavioral disorder, poststroke fatigue, which is clearly distinct from depression in most instances. It is especially disabling and frustrating in that it typically involves patients with total or near-total neurological recovery, who should have been able to go back to their previous activities but who become severely disabled because of early and persisting exhaustion. Preliminary neuropsychological and MR and PET imaging studies suggest that disruption of subtle mechanisms underlying attention, in the absence of significant cognitive and mood alterations, may be responsible. Research projects are now being launched to better delineate poststroke fatigue and its management. PMID- 12649524 TI - New research in the field of stroke: therapeutic hypothermia after cardiac arrest. PMID- 12649527 TI - Patterns of cortical oxygen saturation changes during CO2 reactivity testing in the vicinity of cerebral arteriovenous malformations. AB - BACKGROUND AND PURPOSE: The aim of this study was to test the hypothesis that patterns of cerebrovascular reactivity (CVR) in the vicinity of cerebral arteriovenous malformations (AVMs) before and after resection are not specific for this disease. METHODS: With a microspectrophotometer, cortical oxygen saturation (So2) was measured under steady-state conditions (Paco2, 33 mm Hg) before and after removal of 22 AVMs and in 30 control subjects before and after transsylvian amygdalohippocampectomy. Intraoperative vasoreactivity tests were performed by induced changes of end-tidal CO2 (25, 45, and 25 mm Hg) with simultaneous recording of local So2 in all patients. CVR patterns were established by linear regression analysis (P<0.05) to define parallel (positive) versus inverse (negative) behavior, and reactivity indexes were calculated to define their degree. RESULTS: Cortical oxygenation under steady-state conditions increased significantly (P<0.05) from preoperative to postoperative levels equally in both groups (preoperative AVM, 54.8+/-10.4%So2; postoperative AVM, 73.1+/-10.1%So2; preoperative control, 52.7+/-9.1%So2; postoperative control, 73.6+/-8.9%So2). The rate of inverse CVR patterns increased significantly (P<0.05) from before to after resection without showing statistically significant differences between groups. CONCLUSIONS: Local CVR patterns on presumably normal human cortex of control subjects are heterogeneous, including inverse behavior, and are similar to those of AVM patients before surgery. After surgery, cortical hyperemia is present in both groups, and a significant increase in inverse reactivity patterns interpreted as microvascular steal is noted. An AVM-specific CVR pattern could not convincingly be proved. PMID- 12649526 TI - Cervical sympathetic block to reverse delayed ischemic neurological deficits after aneurysmal subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: The purpose of the present study was to evaluate the feasibility and safety of a locoregional cervical sympathetic block to improve cerebral perfusion in patients suffering from cerebral vasospasm after aneurysmal subarachnoid hemorrhage. METHODS: Nine consecutive patients with symptoms of delayed ischemic deficits, induced by angiographically confirmed cerebral vasospasm, were treated with the injection of locoregional anesthesia to block the ascending cervical sympathetic chain at the level of the superior cervical ganglion. Neurological status was recorded before and after the procedure, and cerebral angiography was performed before and after the procedure. RESULTS: No complications occurred in this short series. The procedure appeared to be simple and safe. Horner's signs appeared within 12+/-0.1 minutes and lasted for an average of 6.3+/-4 hours. In all patients, improved cerebral perfusion was detected at the confirmatory angiography but without change in vessel caliber. One patient died of the complications of the initial hemorrhage, and 2 died of the consequences of the severe vasospasm despite maximal medical treatment. In all the other cases, the neurological status promptly returned to normal within 48 hours after the locoregional treatment. CONCLUSIONS: Patients with mild to moderate symptoms seem to benefit greatly from transient ipsilateral cervical sympathetic block. This simple technique may be helpful when used as an adjunct to the standard therapy to improve cerebral perfusion. PMID- 12649528 TI - A comparison of skating economy on-ice and on the skating treadmill. AB - The purpose of this study was to compare skating economy and oxygen uptake (VO2) on-ice and on the skating treadmill (TM). Male varsity hockey players (n = 15, age = 21.0 yr) performed skating tests on a TM and on-ice. The subjects skated for 4 min at each of 3 submaximal velocities (18, 20, and 22 km . h(-1)), separated by 5 min of passive recovery. A VO2max test followed the submaximal tests and commenced at 24 km . h(-1) with the velocity increasing by 1 km . h(-1) every minute until volitional fatigue. VO2 was 39.7, 42.9, 46.0, and 53.4 ml . kg(-1) . min(-1) at 18, 20, 22, and maximum speed (km . h(-1)) on the TM. VO2 was significantly lower (p < .05) 31.5, 36.9, and 42.7 ml . kg(-1) . min(-1) at 18, 20, and 22 km . h(-1) on-ice. The on-ice VO2max (54.7 ml . kg(-1) . min(-1)) was similar to TM. Stride rate, stride length and heart rate (HR) were significantly different on-ice compared to TM. These results show that at submaximal velocities, VO2, HR, and stride rate are higher on TM compared to on-ice. VO2max was similar while HRmax was higher on the skating treadmill compared to on-ice PMID- 12649529 TI - Epidemiology of chronic kidney disease in Italy: possible therapeutical approaches. AB - Chronic kidney disease (CKD) affects a significant percentage of the Italian population, particularly among the elderly. It is estimated that more than 300 patients per million population (pmp) are diagnosed as having CKD each year, and about 0.8% of Italians are thought to have serum creatinine levels >=1.5 mg/dL. The number of patients being admitted to renal replacement therapies (RRT) has been growing up rapidly in the last decades, leading to 134 patients pmp starting RRT throughout 2000 and to 804 patients pmp on chronic RRT in the same year. As such therapies are very expensive, CKD must be therefore considered as a striking problem also by a socio-economical point of view. As a consequence, any medical intervention being able to halt or at least to slow down the progression of CKD and/or to prevent the development of related complications or comorbidities is of paramount importance. Several therapeutical interventions, including hypertension and proteinuria control, protein restriction, anemia, calcium-phosphate disorders and dyslipidemia correction and smoking cessation, showed to be actually effective in at least partially achieving these objectives. Other emerging therapeutical approaches, although well promising, need further evidence to be definitively included in the management of CKD patients. Particular efforts should be made in order to refer these patients to the nephrologist as early as possible, as it has been widely demonstrated that an early and regular nephrological care leads to decreased morbidity and mortality and also to decreased social costs. PMID- 12649530 TI - New markers of accelerated atherosclerosis in end-stage renal disease. AB - Over the last two decades, several studies have reported a high prevalence of cardiovascular disease in patients with end-stage renal disease (ESRD). This population usually presents both the traditional and non-traditional risk factors for atherosclerosis. Inflammation as well as impaired nitric oxide production are pivotal, throughout the whole process of development of atherosclerotic lesions from the very start. C-reactive protein (CRP), a marker of systemic inflammation and an independent predictor of cardiovascular mortality in the general population, and asymmetric dimethylarginine (ADMA), an endogenous inhibitor of NO synthase, are important risk factors for cardiovascular disease and mortality in the ESRD population. Increased CRP levels have been described in hemo-dialysis and peritoneal dialysis patients, probably due to concomitant diseases, recurrent infections and chronic dialytic therapy. CRP levels, however, are elevated even in predialysis patients, implying that factors related to uremia per se can promote CRP synthesis. Recent reports raise the question whether CRP could be more than just a sensitive marker of inflammation and may contribute actively to the development of the atherosclerotic lesion. ADMA accumulation in the ESRD population is a consequence of reduced renal excretion and impaired enzymatic degradation and is related to the progression of atherosclerosis. Both CRP and ADMA have been shown to be associated with increases in the incidence and progression of atherosclerotic lesions in carotid arteries, as evaluated by high resolution Doppler ultrasonography. PMID- 12649531 TI - Uremic toxicity and anemia. AB - Inappropriate erythropoietin production is the main defect responsible for the anemia of chronic renal failure. However, many other factors can contribute. There is support for the existence in uremic serum of substances that can inhibit erythropoiesis and cause hemolysis, but it is still debated how far uremic retention solutes contribute to the pathogenesis of anemia during chronic renal failure. This article looks at the role of uremic toxicity in exacerbating the anemia of chronic renal failure. PMID- 12649532 TI - Phosphate control in chronic uremia: don't forget diet. AB - Control of the phosphate balance is a major concern for chronic dialysis patients and it depends on dietary intake, intestinal binding and dialytic removal. Phosphorus mass transfer through dialysis and new phosphorus binders have been widely investigated, but negligible attention has been given to dietary phosphorus management, because of the problems of poor compliance and conflict with the recommended high protein intake. The nutritional target in dialysis patients should be a diet supplying adequate protein but limited phosphate intake, without dramatic changes of dietary habits and lifestyle. It is important to educate patients regarding phosphorus content of current foods so that foods providing less phosphorus with the same protein content can be selected, thus preventing dietary phosphate overload. On the basis of a three-day dietary record, dieticians should give the patient personalised advice in order to reduce phosphorus intake while ensuring the desired protein and energy intake. Dietary manipulation may have little impact on the dialysis population but in individual patients dietary counselling can greatly improve phosphate control. Close co operation between nephrologists and dieticians is needed to motivate patients and ensure compliance, if dietary intervention is to succeed. All patients should be given dietary education and counselling, especially young-adults, because dietary phosphate control is an important component of an integrated therapeutic approach to phosphate retention and hyperphosphatemia in end-stage renal disease. PMID- 12649533 TI - Total hip arthroplasty in chronic dialysis patients in the United States. AB - BACKGROUND: The national incidence of and factors associated with total hip arthroplasty (THA) in chronic dialysis patients has never been reported. METHODS: We therefore performed an historical cohort study of 375,857 chronic dialysis patients listed in the 2000 United States Renal Data System between 1 April 1995 and 31 December 1999 and followed-up until 14 May 2000. Primary outcomes were associations with hospitalizations for a primary discharge code of THA (ICD9 procedure code 81.51x) after initiation of dialysis. RESULTS: Dialysis patients had a cumulative incidence of THA of 35 episodes/10,000 person-years, compared to 5.3/10,000 in the general population. The leading indication for THA was osteoarthritis of the hip and pelvis (58% of cases). However, the strongest risk factor for THA in dialysis patients was end-stage renal disease (ESRD) due to systemic lupus erythematosus (SLE, adjusted rate ratio (ARR), 6.80, 95% CI 4.62 10.03, in whom avascular necrosis of the hip was the most common indication, 68.4%). The database did not include information on use of corticosteroids. Diabetic recipients were significantly less likely to receive THA, as were males, and African Americans. Mortality after THA was 0.25% at thirty days and 30% at three years, not significantly different from the expected mortality of dialysis, adjusted for comorbidity. CONCLUSIONS: The most common indication for THA in dialysis patients is osteoarthritis of the hip, similar to the general population. Patients with SLE are more likely to receive THA which is well tolerated and not associated with increased mortality in this population, perhaps reflecting selection bias due to appropriate screening for this elective procedure. PMID- 12649534 TI - Body mass index and enrollment on the renal transplant waiting list in the United States. AB - BACKGROUND: Previous studies of body mass index (BMI) and enrollment on the renal transplant waiting list have not analyzed a national population of chronic dialysis patients. METHODS: 161,265 patients in the United States Renal Data System (USRDS) initiated on end-stage renal disease (ESRD) therapy between 1 April 1995 and 30 June 1997 who had data sufficient to calculate BMI were analyzed in an historical cohort study. Cox Regression analysis was used to model the association of BMI and rates of listing for renal transplantation, censored for death and receipt of renal transplant, and adjusted for demographics, comorbidities in the Medical Evidence Form (2728) and in comparison with patient survival. RESULTS: In univariate analysis, only patients with BMI <21.3 kg/m(2) had a statistically significant lower rate of listing compared to patients with BMI 24.5-28.6 kg/m(2) (6.1 vs. 7.5/100 person years). In Cox Regression analysis, patients with BMI <21.3 kg/m(2) had a 18% lower adjusted rate of listing and a 38% lower rate of survival, and patients with BMI >=24.5-28.7 had an 11% higher adjusted rate of listing and no significant difference in adjusted survival compared with patients with BMI>=28.7 kg/m(2). Caucasian females were disproportionately affected by high BMI. CONCLUSIONS: Among ESRD patients, lower BMI was independently associated with reduced adjusted rates of listing. High BMI affected rates of listing disproportionately in caucasian females. PMID- 12649535 TI - Hospitalized gastrointestinal bleeding and procedures after renal transplantation in the United States. AB - The risk of hospitalized gastrointestinal bleeding (GIB) in renal transplant recipients has not been studied in a national renal transplant population. Therefore, 42,906 renal transplant recipients in the United States Renal Data System (USRDS) from 1 July 1994 - 30 June 1998 were analyzed in an historical cohort study of hospitalizations with a primary discharge diagnosis of GIB (ICD9 Code 578.9x) using Cox regression analysis. The 1997 National Hospital Discharge Survey was used to obtain rates of GIB for the general population. Renal transplant recipients had a cumulative incidence of hospitalizations for GIB of 334 events/100,000 person-years. In 1997, compared to the general population, renal transplant recipients had an age-adjusted rate ratio for GIB of 10.69 at one year of follow-up. The strongest risk factors for GIB in Cox regression analysis were graft loss (adjusted hazard ratio, 4.28 (2.84-6.47) and African American recipients who experienced allograft rejection (AHR, 3.04, 95% CI, 1.45 6.37). GIB was associated with increased all-cause mortality (hazard ratio 1.78, 95% CI, 1.39-2.28). GIB is significantly more common in renal transplant recipients than in the general population, and the strongest risk factors are graft loss and African Americans who experience rejection. PMID- 12649536 TI - Intravascular Doppler technique for monitoring renal venous blood flow in man. AB - BACKGROUND: To measure renal blood flow (RBF) from the renal veins in men using the intravascular Doppler technique (IVD). METHODS: In nine hypertensive male patients (age 46-64 years) undergoing diagnostic renal artery angiography and renal vein catheterization to determine plasma renin activity (PRA), a 3F Doppler catheter was positioned in the renal veins using a 7F guide catheter with a "basket" shaped tip. The radiopaque sectors of the catheter, leaning against the vessel wall, serve to measure the internal diameter of renal veins, and therefore to calculate RBF, by multiplying renal vein cross-sectional area by mean blood flow velocity. The resulting RBF from the left and right renal veins were compared with those obtained by the local thermodilution method (TD). RESULTS: We found good agreement (Bland and Altman's method) between the RBF measurements made with IVD (ranging from 46 mL/min to 1,220 mL/min) and with the TD technique (45-1,030 mL/min) (mean bias, 13+/-20 mL/min, 95% CI -54.77 to 28.77 mL/min). In stenotic kidneys a significant correlation was found between the renal vein PRA and RBF calculated with both methods (IVD: r = 0.96, p = 0.002; TD: r = 0.90, p = 0.01). CONCLUSIONS: The IVD technique applied to the venous side of the renal circulation provides a simple and reliable method for separate measurement of RBF in kidneys with and without renal artery stenosis. PMID- 12649537 TI - Attenuation of renal ischemia/reperfusion injury by a triple drug combination therapy. AB - BACKGROUND: Renal ischemia is of great clinical interest because of its role in renal failure and renal graft rejection. The purpose of this study was to investigate the therapeutic effects of a combination therapy of: n-acetyl cysteine (NAC), a potent antioxidant, sodium nitroprusside (SNP), a nitric oxide donor and phosphormidon (P), an endothelin-1 converting enzyme inhibitor, on tissue protection against renal ischemia/reperfusion injury in the canine model. METHODS: In this study, 15-20 kg male dogs were subjected to 90 minutes of warm unilateral renal ischemia after removal of one kidney and then divided into control, ischemia alone and treatment groups. Blood samples were collected from these dogs for measurement of kidney function tests and the kidneys were harvested at different time intervals for morphological evaluation, immunostaining and Tunnel Assay. RESULTS: Kidney function tests (serum creatinine and blood urea nitrogen [BUN]) showed a significant difference between untreated and treated groups. ** P <0.01, * P< 0.0001 for treated versus untreated. The protective effect of the combination therapy is also supported by light microscopic studies, immunostaining of renal tissue sections for pro-inflammatory cytokines (TNF-alpha and IFN-gamma), iNOS, and apoptosis by TUNEL assay. CONCLUSIONS: Our results suggest that pre-administration of a combination of NAC, SNP, and P attenuates renal ischemia/reperfusion injury. This has potential application in preservation of donor kidney for transplantation by protecting cells against free radical damage. PMID- 12649538 TI - Protective effect of interferon-alpha on carbon tetrachloride-induced nephrotoxicity. AB - BACKGROUND: The aim of this study was to investigate the effect of chronic administration of interferon-alpha 2b for the prevention of carbon tetrachloride (CCl4)-induced oxidative stress and nephrotoxicity. METHODS: Thirty rats were divided into three groups: control, CCl4+placebo (CCl4+P) and CCl4+interferon alpha 2b (CCl4+INF). Control rats were treated with pure olive oil. The other rats were treated for seven weeks with subcutaneous injections of CCl4 (0.15 mL /kg) in pure olive oil three times a week. Rats were killed at the end of the seventh week and renal histopathological examinations were done: specimens of renal tissue were obtained for investigating oxidative stress parameters, including malondialdehyde (MDA) and glutathione peroxidase (GSH-Px). RESULTS: Tubular changes, glomerular hypercellularity, and capillary obliteration were significantly less in the CCl4+INF group than with CCl4+P (p<0.05) and the interstitial fibrosis score for the CCl4+INF group was similar to the control group. However, the interstitial inflammation score was higher in the CCl4+INF group than the control group (p<0.05). No change was observed in the CCl4+P group. Renal MDA levels in the control and CCl4+INF groups were significantly lower than the CCl4+P group, while GSH-Px was significantly higher (p<0.001). There was no difference between the control and CCl4+INF groups in oxidative stress markers (p>0.05). CONCLUSIONS: Administration of interferon-alpha 2b to CCl4-treated rats prevented interstitial fibrosis, probably as a result of its antifibrogenic effect. It also reduced intrarenal oxidative stress in rats with CCl4-induced nephrotoxicity. PMID- 12649539 TI - Inhibition of 5-lipoxygenase activating protein decreases proteinuria in diabetic rats. AB - BACKGROUND: The binding of 5-lipoxygenase (5-LO) to the 5-LO activating protein (FLAP) is a prerequisite for subsequent formation of leukotrienes (LT) from arachidonic acid. We have shown that FLAP antagonist administration decreased proteinuria in glomerulonephritic patients. In this follow-up study, we assessed the role for FLAP in a rat model of streptozotocin-induced diabetic nephropathy. METHODS: Diabetic rats were treated for 4 weeks with FLAP (BAY X-1005, 200 mg/kg) or 5-LO (Zileuton, 80 mg/Kg) antagonists. Proteinuria, renal function and LT production was assessed. We also determined protein permeability of cultured glomerular endothelial cells (which possess no 5-LO) by measuring their permeability to radiolabeled albumin with and without FLAP antagonists. RESULTS: FLAP mRNA levels increased dramatically in glomeruli from diabetic animals compared to controls. Inhibition of FLAP (but not inhibition of 5-LO) reduced proteinuria, with no effect on estimated glomerular filtration rate. Interestingly, diabetes-induced rises in urinary excretion and glomerular production of leukotrienes were not modified by the inhibitors. Increased FLAP expression in glomerular endothelial cells in culture was associated with an increase in albumin permeability, and this increase was abolished by FLAP antagonists. On the other hand, addition of LTA(4) led to increases in leukotriene formation and in permeability. This increase in permeability was also reduced by co-incubation with FLAP antagonists, whereas the increase in leukotriene synthesis was not modified. CONCLUSIONS: These results suggest a role for FLAP other than the activation of 5-LO, possibly in protein handling, and point to FLAP antagonists as anti-proteinuric agents. PMID- 12649540 TI - Involvement of TGF-beta signal for peritoneal sclerosing in continuous ambulatory peritoneal dialysis. AB - BACKGROUND: Functional failure of the peritoneal membrane is the most serious problem in long-term continuous ambulatory peritoneal dialysis (CAPD). Transforming growth factor-beta (TGF-ss) is one of the key mediators of fibrosis in some organs, and is thought to be involved in peritoneal alterations. In this study, we examined the role of TGF-beta1/TGF-ss receptors for human peritoneal mesothelial cells (HPMCs) and fibroblasts, and their interactions in CAPD patients. METHODS: HPMCs were cultured for 48 h in a medium containing normal- dose glucose (7 mM), high-dose glucose (30 mM) and mannitol as an osmotic agent, equal to 30 mM glucose. Cell proliferation was observed using the Tetra Color One assay. The concentration of TGF-beta1 in culture supernatants was measured by enzyme-linked immunosorbent assay (ELISA). The expression of TGF-ss receptor types I and II was observed by flow cytometry. HPMCs and fibroblasts were co cultured and assayed using transwell inserts in order to identify the effects of the high-concentration glucose solution. RESULTS: HPMC proliferation was inhibited by the high concentration of glucose but not by mannitol. The inhibition was abrogated by the neutralizing antibody for TGF-beta1. TGF-beta1 was induced by a high concentration of glucose but not by mannitol. The expression of both TGF-ss receptors was augmented in culture with the high concentration of glucose but not with mannitol. In the co-culture assay, the number of HPMCs was decreased and fibroblasts were significantly increased in culture with the high concentration of glucose. CONCLUSIONS: A high concentration of glucose induced a large amount of TGF-beta1 and enhanced the expression of TGF ss receptors. HPMCs were sensitive to TGF-beta1 in response to a high concentration of glucose. These data suggest that TGF-beta1 from HPMCs exposed to a high concentration of glucose down-regulates the proliferation of HPMCs and accelerates peritoneal fibrosis. PMID- 12649541 TI - Regulation of glucose transporters in human peritoneal mesothelial cells. AB - BACKGROUND: Risk factors for peritoneal fibrosis and mesothelial cell (MsC) injury in CAPD are infections and bioincompatibility of the dialysate, including high glucose concentrations. To study a potential link between dialysate and glucose toxicity in MsC, we investigated the expression of facilitative glucose transporters (GLUT), which could contribute to glucose toxicity. METHODS: After induction of cell differentiation, MsC were incubated in regular medium or medium with 60 mM D-glucose, 30 mM glucose plus 30 mM mannitol, 60 mM mannitol, PD effluent, or with a cytokine mix. Expression of GLUT1, GLUT3, SGLT and GAPDH/L32 was studied by RNase protection assay. MsC were incubated under identical conditions with 14C-fluoro-deoxy-glucose for 30 minutes and glucose uptake was measured. To estimate Vmax and Km, 14C-fluoro-deoxy-glucose uptake rates were determined over a range of 0.6 to 10 mM unlabeled glucose. RESULTS: The cytokine mix significantly stimulated GLUT1 expression (3-fold) and GLUT3 (1.7-fold). There was a 1.4-fold increase in GLUT1 (p<0.05) and a 1.7-fold increase in GLUT3 (p<0.05) after incubation in high glucose but not in mannitol or PD-effluent controls. Glucose uptake studies confirmed this increase after incubation in 30 mM (p<0.05) and 60 mM glucose solutions. Kinetic studies showed the Km was approximately 3.7 mM for this transport. CONCLUSIONS: GLUT mRNA expression and glucose uptake are induced by high ambient glucose concentrations and cytokines. Unlike many other cells, MsC are not able to protect themselves from increased glucose concentrations by downregulation of GLUTs. The intracellular glucose concentration may therefore increase during CAPD, affecting growth factor expression and glycosylation, and contributing to glucose toxicity. PMID- 12649542 TI - BB genotype of the vitamin D receptor gene polymorphism postpones parathyroidectomy in hemodialysis patients. AB - BACKGROUND: Bsml vitamin D receptor (VDR) gene polymorphism has been reported to influence the progression of secondary hyperparathyroidism but it is not known how much the genetic background contributes to the need for parathyroidectomy (PTx). We investigated the influence of VDR gene polymorphism on PTx in patients with different dialysis vintage. METHODS: We studied 121 parathyroidectomized HD patients ("PTx " group). Patients who had required early parathyroidectomy ("early PTx" group) or late parathyroidectomy ("late PTx" group) were analyzed separately. The cut-off point between these two groups was 89 months (mean time on hemodialysis (HD) before parathyroidectomy). Serum intact parathyroid hormone, calcium, phosphorus and alkaline phosphatase were measured. Bsml genotypes were analyzed by polymerase chain reaction. Statistical analysis was done with univariant analysis of variance (ANOVA) to compare the genotype groups and general factorial ANOVA, entering time on HD as the dependent variable, with genotype, sex, age and chronic renal failure (CRF) etiology as factors. As a control group for the association studies we determined genotypic frequencies in 162 HD patients ("total HD" group), and in a healthy control population of 120 individuals ("healthy" group), tested by contingency table analysis and the chi square test. RESULTS: No significant differences were found between the genotypes except for the time on HD. General factorial ANOVA showed that the adjusted means of the time on HD were significantly different for the various genotypes (p = 0.015). The BB genotype was significantly less frequent in the "early PTx " group than in the "total HD" and "late PTx" groups. CONCLUSIONS: Individuals with the BB genotype can remain longer on HD before they need parathyroidectomy. PMID- 12649543 TI - Effect of erythropoietin therapy on polyneuropathy in predialytic patients. AB - BACKGROUND: Peripheral neuropathy commonly develops in patients with advanced chronic renal failure. The uremic neuropathy is often subclinical and detectable only by electrophysiological studies. Receptors to erythropoietin (EPO) have been described on non-hematopoietic cells including neuronal cells. METHODS: In order to evaluate the effect of five months' EPO therapy on polyneuropathy in predialytic patients, nerve conduction studies (NCS) were done in 46 anemic predialytic patients without neurological complaints. In 22 (twelve non-diabetic and ten diabetic) neuropathy was detected and these patients were included in the study. After five months of subcutaneous EPO therapy NCSs were repeated. RESULTS: Hemoglobin increased significantly (p=0.0001) with no significant increase in plasma creatinine. Motor nerve conduction velocity (MNCV) and compound muscle action potentials (CMAP) of the ulnar nerve were normal before EPO therapy and at the end of the study. MNCV of the median, peroneal and tibial nerves improved significantly (p<0.05). CMAP of the median nerve rose significantly, to the normal range (p=0.01). Sensory nerve conduction velocity (SNCV ) and sensory nerve action potentials (SNAP) were reduced in all sensory nerves and did not improve. The improvement in non-diabetic patients was better than in diabetic patients. No significant correlation was found between the increase in hemoglobin and the improvement in MNCV. CONCLUSIONS: Subcutaneous EPO therapy improved motor polyneuropathy in uremic patients, especially non-diabetic individuals. The improvement in MNCV may reflect remyelination. This non-hematopoietic effect may be related to some direct action through EPO receptors on peripheral neuronal cells. PMID- 12649544 TI - Behavioral and functional abnormalities linked with recurrent urinary tract infections in girls. AB - BACKGROUND: Most girls with recurrent urinary tract infections do not have major urinary tract abnormalities. Recent studies focus on predisposing behavioral and functional abnormalities: infrequent voiding, inadequate fluid intake, stool retention, poor genital hygiene and voiding dysfunction. METHODS: Complete history, bladder and bowel questionnaire, physical examination, voiding-drinking diary, sonography and uroflowmetry were used to assess infrequent voiding, functional stool retention, poor fluid intake, inadequate hygiene, or voiding dysfunction in girls referred for evaluation of three or more symptomatic urinary tract infections (with a first infection at the age of more than 36 months). RESULTS: A total of 141 girls aged 3.9 to 18 years were evaluated between 1996 and 1999; 212 abnormalities were noted in 120 patients: infrequent voiding (isolated, 16; combined with other abnormalities, 47), poor fluid intake (isolated, 10; combined, 50), functional stool retention (isolated, 5; combined, 25), inadequate hygiene or toilet habits (isolated, 3; combined, 24), dysfunctional voiding (isolated, 15; combined, 10), bladder overactivity (isolated, 5; combined, 2). CONCLUSIONS: Most girls referred for evaluation of three or more urinary tract infections have host-mediated predisposing abnormalities: infrequent voiding, poor fluid intake, functional stool retention or voiding dysfunction. Poor genital hygiene and toilet habits were almost always combined with other abnormalities, suggesting that infections are not necessarily related to poor genital hygiene or toilet habits. Two or more indications of predisposing behavior often concur in the same patient. PMID- 12649545 TI - Cholesterol crystal embolic disease in renal allografts. AB - Cholesterol embolic disease in the renal allograft is not recognized as an important cause of graft dysfunction. We describe here two renal transplant patients with cholesterol embolization in their allograft biopsies. The first, a 48-year-old patient, received a renal transplant from a 62-year-old donor with a history of hypertension and tobacco use. On account of initial non-function, a renal biopsy was taken, which showed acute tubular necrosis and cholesterol emboli. The second, a 55-year-old man, presented chronic allograft failure six years after transplantation; ultrasonography showed a solid renal mass. Nephrectomy specimens revealed renal carcinoma and a combination of chronic rejection and multiple cholesterol emboli. Cholesterol embolic disease is probably an under-reported cause of renal graft dysfunction. The source of the emboli may be either the donor or the recipient's vessels. Since the current tendency is to accept older donors and recipients with more advanced atherosclerotic disease, this condition is likely to become more frequent in the future. Particular care must be taken at the time of organ procurement and during the evaluation of organ donors, in order to reduce the risk of embolization. PMID- 12649546 TI - Kidney transplant in Gitelman's syndrome. Report of the first case. AB - This paper is the first report of a patient with Gitelman's syndrome who, after developing a chronic nephropathy leading to end-stage renal disease, underwent kidney transplantation. The clinical findings of this disease, which include hypokalemia, high angiotensin II and aldosterone levels, sustained hyporesponsiveness to the pressor action of angiotensin II and norepinephrine, normo/hypotension and hypovolemia; the clinical course after kidney transplantation highlights the importance and the need for carefully controlling the hemodynamic status of these patients. In fact, the persistence of normo/hypotension and hypokalemia, in the presence of increased levels of Ang II and aldosterone to which the transplanted kidney should normally respond, raises interesting questions about the mechanisms responsible for the regulation of patient's vascular tone and potassium homeostasis after transplantation, which could expose the patient to post-transplant hypoperfusive renal failure and its long-term complications. PMID- 12649547 TI - Unexpected IgA nephropathy during the treatment of a young woman with idiopathic dermatomyositis: case report and review of the literature. AB - This article reports the unexpected discovery of IgA nephropathy in a 26-year-old Chinese woman 1.5 years after the onset of idiopathic dermatomyositis. The patient was taking immunosuppressive agents, prednisolone 25 mg and azathioprine 75 mg daily. Glomerulonephritis associated with idiopathic polymyositis/dermatomyositis is rare. A review of the medical literature indicates that the most common pattern seen in idiopathic polymyositis is mesangial proliferative glomerulonephritis. However, both membranous and mesangial proliferative glomerulonephritis are often seen in idiopathic dermatomyositis. It is still not clear, however, whether the humorally- mediated immune process in dermatomyositis and the cell-mediated immune process in polymyositis can explain the different patterns of occurrence of glomerular lesions in these two closely related disease entities. PMID- 12649548 TI - IgA deficiency with membranous glomerulonephritis: a case report and review. AB - Selective immunoglobulin A (IgA) deficiency may result in a predisposition to recurrent sinopulmonary infection and allergic diseases. IgA deficiency may also play a role in the development of autoimmune disorders. Selective IgA deficiency associated with glomerulonephritis was rare, while the clinical presentation in IgA deficiency-associated glomerulonephritis was variable. We report an 83 year old male with selective IgA deficiency associated with membranous glomerulonephritis. He presented with nephrotic syndrome. Percutaneous renal needle biopsy showed diffuse global thickening and rigidity of glomerular capillary walls, mildly diffuse segmental expansion of mesangial matrix, focal and cortical scar with segmental obsolescence of glomeruli. Heavy IgG and moderate C3 deposits were found on immunofluorescence. We also review the previous cases of IgA deficiency with glomerulonephritis. Several clues were rendered to establish the association between IgA deficiency and membranous glomerulonephritis. PMID- 12649549 TI - Salt restriction and not length of dialysis is the key to drug free blood pressure control in ESRD patients. PMID- 12649550 TI - Temkin's times and ours: an appreciation of Owsei Temkin. PMID- 12649551 TI - Who was Akilaos? A problem in medical historiography. AB - In the years before the conquest of Alexandria by the Arabs in 642, Alexandrian scholars produced compendia, summaries, and commentaries on Galen's writings and other works in the Hippocratic medical tradition. This paper concerns the identities of those Alexandrian commentators-two in particular, whose names are reported in Arabic texts, but about whom little else is known-and the routes by which Galenic medicine was transmitted to other parts of the Mediterranean world after Alexandria's fall. We review debates over the identity of "Akilaos" and "Anqilaos," and argue that "Akilaos" is not merely a scribal duplication, as some have supposed. Instead, we suggest a connection with the city of Aquileia in northern Italy, as a source for the name "Akilaos" and as a route by which Alexandrian medical texts reached northern Italy (where their Latin versions subsequently were associated with a "Ravenna school" of medicine). PMID- 12649552 TI - Serving Clio and client: the historian as expert witness. AB - Although historians often appear in court as expert witnesses, their presence stirs unease and controversy. To clarify the issues at stake, this article compares two activities-testifying on behalf of plaintiffs, and conducting an open-ended historical inquiry-by using the author's personal experience in Craft v. Vanderbilt as a case in point. The litigation sought to gain compensation and an apology for the 830-850 women who between 1945 and 1949 at the Vanderbilt prenatal clinic were fed doses of radioactive iron without their consent so as to study the process of iron absorption. The overall conclusion is that historians can serve clients without subverting the canons of the discipline. However, because Clio and client have such different needs, historians should recognize, and take pride in the fact, that courtroom appearances represent advocacy. PMID- 12649553 TI - The new Poor Law guardians and the administration of insanity in East London, 1834-1844. AB - One of the main aims of the Poor Law Amendment Act of 1834 was to impose national consistency of practice in poor relief. Central guidance was designed to produce uniformity in guardians' policies on dealing with the insane. This study of the administration of insanity in the eastern metropolis in the first decade after the Act demonstrates that the new boards of guardians were as culturally distinctive in their style of administration of the new Poor Law as the parishes had been under the old regime. A complex interplay of personality, politics, and class determined the corporate culture of individual boards. PMID- 12649554 TI - Seeds of discord: the politics of radon therapy in Canada in the 1930s. AB - In the early twentieth century, the therapeutic use of radon gas became an accepted medical practice. "Radium emanation" plants were established in many parts of North America to supply radon seeds to physicians. In Canada, these plants were usually established as part of state-supported cancer programs, creating concern among the medical profession, which had hitherto directed cancer treatment. This article explores how issues surrounding the ownership and distribution of radon played out in two Canadian provinces, Manitoba and Ontario. The main focus is an analysis of a computerized database created from more than two thousand radon order forms, dating from 1933 to 1940, preserved in the Archives of Ontario, which reveals interesting information about patients and the uses of radon in the 1930s, as well as discrepancies between policy and practice that illuminate the medical politics of the era. Although the radon seeds were intended for use in the government-supported central cancer clinics, they were widely distributed to practitioners throughout Ontario, and many patients received treatment for noncancerous conditions. These discrepancies are explored in the context of the struggles over cancer policy between the government and the Ontario medical profession. The article also shows how similar conflicts evolved in Manitoba. Finally, the distribution of radon is linked to the public acceptance of medical radiation despite contemporary reports of harm. PMID- 12649555 TI - "What do these women want?": Feminist responses to Feminine Forever, 1963-1980. AB - In 1963, Brooklyn gynecologist Robert A. Wilson and his wife, Thelma, published a paper in the Journal of the American Geriatrics Society arguing that untreated menopause robbed women of their femininity and ruined the quality of their lives. In 1966 Robert Wilson published a best-selling book, Feminine Forever, in which he maintained that menopause was an estrogen-deficiency disease that should be treated with estrogen replacement therapy to prevent the otherwise inevitable "living decay." This paper explores the issues raised by the convergence of Wilson's campaign and the emergence of the women's movement. Between 1963 and 1980, feminists did not respond with one voice to Wilson's ideas: at first, some embraced them as a boon for aging women, while others resisted regarding female aging as pathological. In 1975, studies linking ERT and endometrial cancer challenged the wisdom of routine hormone therapy; this shifted the tenor of the feminist discussion, but it did not create a consensus about the meaning of menopause or its treatment. Nevertheless, the feminist discussion of menopause revealed a larger women's health agenda-namely, the unyielding belief that women should retain control of their bodies and participate fully in the decision making efforts regarding their health. By controlling their bodies, all women, whether feminist or not, could ultimately control their lives. PMID- 12649556 TI - Pregnant children and half-dead adults: modern living and the quickening life cycle in Botswana. AB - This paper utilizes ethnography and oral history to examine local perspectives on one aspect of the health transition, the female life cycle, in postcolonial Botswana. Botswana has undergone a remarkably rapid epidemiologic transition in recent decades, and it thus provides a unique context within which local actors analyze the interaction between biological and sociocultural change. Improvements in the standard of living have resulted in both an earlier onset for puberty in girls and an increased incidence of stroke among older women, thus refashioning the female life course. Local analysis and commentary on the shifting norms of women's bodies read this phenomenon alongside broader historical transformations. In the process they complicate basic assumptions in international health about the meanings of health and development. PMID- 12649559 TI - Incidence, etiology, and impact of Fever in patients with acute pancreatitis. AB - BACKGROUND/AIM: Fever, as a significant event, has not been studied systematically in patients with acute pancreatitis. We studied prospectively incidence, etiology, and impact of fever on the management and outcome in patients with acute pancreatitis. METHODS: All consecutive patients with acute pancreatitis were studied for the development of fever, its etiology, and its influence on the management and outcome of acute pancreatitis. Fever was considered to be significant, if the temperature was >38 degrees C and persisted for >2 days. RESULTS: A total of 75 patients (51 males; mean age 41 years) with acute pancreatitis were included between January 1997 and June 1998. The causes of pancreatitis were gallstones in 48%, alcohol in 28%, and others in 24% of the patients. 20 patients had pancreatic necrosis, and 45 (60%) developed fever during the course of pancreatitis. The etiology of fever was infected pancreatic necrosis in 8 (18%), pancreatitis per se in 10 (22%), cholangitis in 4 (9%), nonpancreatic infections in 17 (38%), and an undetermined one in 6 (13%) patients. Of the 45 patients with fever, 17 had pancreatic necrosis as compared with only 3 of 30 patients who did not develop fever (p < 0.05). Patients with fever had a higher pancreatitis-related mortality than those without fever (p = 0.03). CONCLUSIONS: 60% of the patients with acute pancreatitis developed fever. Infected pancreatic necrosis was the cause of fever in 18% of the patients and not in the majority, i.e., 82% of the patients. The mortality rate was higher in patients who developed fever than in those who did not. PMID- 12649560 TI - Multimodal management - of value in fulminant acute pancreatitis? AB - BACKGROUND: The multiple organ dysfunction syndrome (MODS) is the major cause of morbidity and mortality associated with acute pancreatitis. Presently, therapy is merely organ supportive as no effective therapy against underlying causative pathophysiological mechanisms exists. AIMS: To evaluate the effect of treatment with a platelet-activating factor inhibitor (PAFI), a monoclonal antibody against platelet endothelial cell adhesion molecule 1 (PECAM-1-MAb) and an oxygen free radical scavenger (N-acetylcystein; NAC), alone or in combination, on systemic organ dysfunction in experimental acute pancreatitis. METHODS: Severe acute pancreatitis was induced in rats by the intraductal administration of taurodeoxycholate. Treatment was given after 1 or 3 h, and evaluations were performed 6 h after induction. Organ dysfunction was evaluated by means of endothelial integrity impairment expressed as endothelial barrier leakage index. RESULTS: Severe acute pancreatitis caused a significant impairment in endothelial integrity in all organs studied and decreased levels of protease inhibitors compared to controls. The endothelial barrier impairment was significantly ameliorated by all treatment modalities, either given early or later. Combinations of NAC and the PECAM-1-MAb or the PECAM-1-MAb and the PAFI were the only schedules to restore endothelial barrier integrity to normal levels in most of the organs studied. CONCLUSION: Combination therapy with NAC and PECAM-1-MAb and/or PAFI may offer effective, causative-directed supplements to organ supportive therapy of MODS in severe acute pancreatitis. PMID- 12649561 TI - Role of PGE2 in the development of pancreatic injury induced by chronic alcohol feeding in rats. AB - BACKGROUND: Eicosanoids are known to modulate inflammation. Moreover, some studies report that endogenous prostaglandin E(2) (PGE(2)) protects the pancreas against injury. Therefore, we investigated its role in a rat model of chronic alcohol consumption. METHODS: Rats were fed with 20% ethanol and a corn oil supplemented diet using the interrupted alcohol feeding regimen (EI). Controls received water instead of ethanol (WI) or uninterruptedly ethanol (EU). After 13 mo, pancreas tissue was investigated morphologically, immunohistochemically and biochemically. RESULTS: Pancreatic tissue was more severely injured in EI than in WI and EU (p < 0.05). Fibrogenesis (alpha-smooth muscle actin-positive cells, collagen types I and III) was increased in EI compared to WI (p < 0.05). In EI, mast cell numbers were increased, compared to WI, but decreased, compared to EU (p < 0.05). EI showed decreased PGE(2) and malondialdehyde contents compared to EU (p < 0.05) and decreased glutathione concentrations compared to WI (p < 0.05). PGE(2) content and fibrogenesis were inversely correlated in EU. The same correlation was detectable as a trend in all alcohol-fed rats. CONCLUSION: The decrease in PGE(2) together with the increase in tissue damage and the inverse correlation between PGE(2) and fibrogenesis led us to suggest that endogenous PGE(2) plays a protective role in alcohol-induced injury in the pancreas. PMID- 12649562 TI - An enteral therapy containing medium-chain triglycerides and hydrolyzed peptides reduces postprandial pain associated with chronic pancreatitis. AB - BACKGROUND/AIM: Pain in patients with chronic pancreatitis is difficult to manage. We examined if an enteral formulation containing medium-chain triglycerides (MCT) and hydrolyzed peptides would (1) minimally stimulate the exocrine pancreas by blunting cholecystokinin release and (2) decrease pain in patients with chronic pancreatitis. METHODS: In the first part of the study, on separate days, 6 healthy controls consumed a standard enteral formulation, an enteral formulation containing MCT and hydrolyzed peptides, and a high-fat meal. Baseline and postprandial plasma cholecystokinin (CCK) concentrations were analyzed. Subsequently, 8 patients with chronic pancreatitis were enrolled and instructed to complete a visual analog pain assessment for a baseline period of 2 weeks followed by three cans per day of the enteral formulation containing MCT and hydrolyzed peptides for 10 weeks. RESULTS: Mean CCK levels for our control subjects were 0.46 +/- 0.29 pM at baseline, 10.75 +/- 0.45 pM in response to the high-fat meal, and 7.9 +/- 1.25 pM in response to the standard enteral formulation. Of note, CCK levels were 1.43 +/- 0.72 pM in response to the enteral supplement containing MCT and hydrolyzed peptides. In patients with chronic pancreatitis, the average improvement in pain scores from baseline to the conclusion of the study was 61.8% (p = 0.01). This corresponded to a clinical improvement in 6 of the 8 patients. CONCLUSIONS: A complete enteral supplement containing MCT and hydrolyzed peptides minimally increases plasma CCK levels. This therapy may be effective in reducing postprandial pain associated with chronic pancreatitis. PMID- 12649563 TI - Vitamin D receptor is expressed in pancreatic cancer cells and a vitamin D3 analogue decreases cell number. AB - BACKGROUND AND AIM: The vitamin D-receptor (VDR) has been detected in both normal and malignant cells of different tissues. Treatment with vitamin D(3) has been suggested as a possible therapy in malignant diseases such as pancreatic cancer. Synthetic analogues of vitamin D(3) have a less hypercalcemic effect than native vitamin D(3). The aim was to study the expression of the VDR in human pancreatic cancers and to study the in vitro effect of an analogue to vitamin D(3) on cell lines established from these cancers. METHODS: The pancreatic cancer cell lines were established from primary cultures with only cancer cells. A probe specific for the human VDR was used. After reverse-transcriptase PCR and Northern blotting, the expression of the VDR in normal pancreas and in pancreatic cancers was compared. The cell lines were incubated with EB 1089, a synthetic analogue vitamin of D(3), in dose-response studies. The cell number was measured by the XTT colorimetric method. RESULTS: The VDR was expressed in all cancers and in six of the cell lines the expression was increased more than 3-fold compared to normal pancreas. All cell lines developed from human pancreatic cancers responded with a decreased cell number to the vitamin D(3) analogue at concentrations of 10(-5) M or higher. CONCLUSION: The VDR was expressed in all pancreatic cancers studied. Cell lines derived from these cancers responded with a decrease in cell number to high concentrations of a vitamin D(3) analogue. These results, and the doses to use, have to be confirmed with in vivo studies. PMID- 12649564 TI - Transient agonist-induced regulation of the cholecystokinin-A and cholecystokinin B receptor mRNA levels in rat pancreatic acinar AR42J cells. AB - BACKGROUND: CCK-8 and gastrin exert multiple effects in the gastrointestinal tract and the nervous system. Their actions are mediated via the G-protein coupled CCK-A and CCK-B receptors. METHODS: Rat pancreatic acinar tumor AR42J cells express both CCK receptor subtypes. This cell line was used to characterize the agonist-dependent regulation of CCK-A and CCK-B receptor gene expression. RESULTS: CCK-8 (10 nM) or gastrin (10 nM) reduced CCK-A receptor mRNA expression to 56% and 53%, respectively 2 h after hormonal exposure. In contrast, the level of CCK-B receptor gene expression was upregulated to 157% and 153%, respectively. These effects are most probably linked to the CCK-B receptor in AR42J cells. The phorbolester PMA (100 nM), a protein kinase C activator, downregulated CCK-A receptor expression but did not affect CCK-B receptor gene transcription. Activation of protein kinase A by forskolin (10 microM) or Bt(2)cAMP (100 microM) is not involved in the transient regulation of CCK receptor mRNA expression. Both elevated CCK-B and decreased CCK-A receptor mRNA expression returned to basal levels 6 h after continuous stimulation. CONCLUSION: These results demonstrate that CCK-A and CCK-B receptor mRNA levels are differentially regulated by their agonists via distinct signal transduction mechanisms in AR42J cells. PMID- 12649565 TI - Intra-arterial instillation of microencapsulated, Ifosfamide-activating cells in the pig pancreas for chemotherapeutic targeting. AB - BACKGROUND: The therapeutic efficacy of intratumoral instillation of genetically engineered, CYP2B1-expressing, microencapsulated cells in combination with ifosfamide had been previously demonstrated in xenografted human pancreatic ductal carcinomas [Gene Ther 1998;5:1070-1078]. Prior to a clinical study, the feasibility of an intra-arterial application of microencapsulated cells to the pancreas and its consequences to the organ had to be evaluated. MATERIAL AND METHODS: Microencapsulated, CYP2B1-producing cells were instilled both in vivo (transfemoral angiographical access) and in vitro (perfusion model) in the splenic lobe of the pig pancreas. In vivo, animals were monitored clinically for 7 days, then treated with ifosfamide and sacrificed. In vitro, ifosfamide was administered intra-arterially. RESULTS: In all animals, 100 microcapsules could be instilled safely via the femoral route without clinical, biochemical or histological signs of pancreatitis. Histological examination revealed partial obstruction of small arteries by the capsules, without causing any parenchymal damage. In vitro, instillation reduced blood flow by half. Ifosfamide, also in combination with the capsules, did not add any damage to the pancreas. CONCLUSION: Intra-arterial instillation of microencapsulated cells to the pig pancreas is feasible and safe. Neither pancreatitis, foreign body reactions nor circulatory disturbances were observed. Clinical application of this genetically enhanced chemotherapeutic method seems possible. PMID- 12649566 TI - Fatal acute pancreatitis. Characteristics of patients never reaching hospital. AB - BACKGROUND: Mortality in acute pancreatitis is still substantial. The extent of patients dying at home due to acute pancreatitis, thus never admitted to hospital, is scarcely known. MATERIAL AND METHODS: Thirty-six patients with death caused by acute pancreatitis were identified at the Department of Forensic Medicine over a 7-year period. RESULTS: Twenty-seven patients (75%) were assumed to have alcohol as the underlying etiological factor. The duration of abdominal pain seemed short, with a median of one day. Profound pancreatitis changes were found in all patients while pulmonary changes were less frequent. CONCLUSION: Overall, up to 1/3 of patients dying from acute pancreatitis in our region seem to die at home never being admitted to hospital. The majority of instances was caused by alcohol abuse. The progression of the cause of the disease seemed rapid, implying that these patients could constitute a specific subgroup of severe acute pancreatitis. PMID- 12649567 TI - N34S, a pancreatitis associated SPINK1 mutation, is not associated with sporadic pancreatic cancer. AB - BACKGROUND: The SPINK1 mutation N34S is associated to chronic pancreatitis of idiopathic, alcoholic and tropic origin. Our aim was to evaluate its prevalence in patients with sporadic pancreatic cancer. METHODS: The N34S allele of SPINK1 was analyzed by melting curve analysis using fluorescence resonance energy transfer (FRET) probes and the LightCycler in 159 German patients with sporadic pancreatic cancer (83 male, 76 female, median age of diagnosis 61 years, range 36 84 years) and 492 healthy controls. RESULTS: The N34S mutation was detected in 2 (1.3%) of the 159 patients with pancreatic cancer and in 8 (1.6%) of the 492 controls (not significant). CONCLUSION: The N34S mutation of SPINK1 appears not to be a distinct genetic risk factor in patients with sporadic pancreatic cancer. PMID- 12649568 TI - VMP1 expression correlates with acinar cell cytoplasmic vacuolization in arginine induced acute pancreatitis. AB - BACKGROUND: Recently, we described the cloning of VMP1 (vacuole membrane protein 1). In vitro expression of VMP1 promotes formation of cytoplasmic vacuoles followed by cell death. In order to test if VMP1 expression is related to the cytoplasmic vacuolization of the acinar cells during acute pancreatitis, we studied the in vivo expression of the new gene during arginine-induced acute pancreatitis. METHODS: Male Wistar rats injected with 500 mg/100 g of L-arginine were time-course sacrificed and pancreas tissue removed. RESULTS: Northern blot analysis showed maximal induction of VMP1 after 24 h remaining high after 48 h of arginine administration. Significant increase in the number of TUNEL-stained cells were found at those periods. After 24 and 48 h of arginine administration, light micrographs from thin plastic toluidine blue sections revealed numerous vacuoles in the cytoplasm of acinar cells. In situ hybridization studies showed high expression of VMP1 in acinar cells with cytoplasmic vacuolization. VMP1 mRNA highly and significantly correlated with vacuole formation. CONCLUSION: These results suggest that VMP1 expression may be involved in the cytoplasmic vacuolization of acinar cells during the early stage of acute pancreatitis. PMID- 12649569 TI - Short note about a symposium on alcoholic pancreatitis. PMID- 12649570 TI - Pediatric stress: hormonal mediators and human development. AB - Stress activates the central and peripheral components of the stress system, i.e., the hypothalamic-pituitary-adrenal (HPA) axis and the arousal/sympathetic system. The principal effectors of the stress system are corticotropin-releasing hormone (CRH), arginine vasopressin, the proopiomelanocortin-derived peptides alpha-melanocyte-stimulating hormone and beta-endorphin, the glucocorticoids, and the catecholamines norepinephrine and epinephrine. Appropriate responsiveness of the stress system to stressors is a crucial prerequisite for a sense of well being, adequate performance of tasks and positive social interactions. By contrast, inappropriate responsiveness of the stress system may impair growth and development, and may account for a number of endocrine, metabolic, autoimmune and psychiatric disorders. The development and severity of these conditions primarily depend on the genetic vulnerability of the individual, the exposure to adverse environmental factors and the timing of the stressful event(s), given that prenatal life, infancy, childhood and adolescence are critical periods characterized by increased vulnerability to stressors. The developing brain undergoes rapid growth and is characterized by high turnover of neuronal connections during the prenatal and early postnatal life. These processes and, hence, brain plasticity, slow down during childhood and puberty, and plateau in young adulthood. Hormonal actions in early life, and to a much lesser extent later, can be organizational, i.e., can have effects that last for long periods of time, often for the entire life of the individual. Hormones of the stress system and sex steroids have such effects, which influence the behavior and certain physiologic functions of individuals for life. Exposure of the developing brain to severe and/or prolonged stress may result in hyperactivity/hyperreactivity of the stress system, with resultant amygdala hyperfunction (fear reaction), decreased activity of the hippocampus (defective glucocorticoid-negative feedback, cognition), and the mesocorticolimbic dopaminergic system (dysthymia, novelty-seeking, addictive behaviors), hyperactivation of the HPA axis (hypercortisolism), suppression of reproductive, growth, thyroid and immune functions, and changes in pain perception. These changes may be accompanied by abnormal childhood, adolescent and adult behaviors, including excessive fear ('inhibited child syndrome') and addictive behaviors, dysthymia and/or depression, and gradual development of components of the metabolic syndrome X, including visceral obesity and essential hypertension. Prenatal stress exerted during the period of sexual differentiation may be accompanied by impairment of this process with behavioral and/or somatic sequelae. The vulnerability of individuals to develop varying degrees and/or components of the above life-long syndrome is defined by as yet unidentified genetic factors, which account for up to 60% of the variance. CRH has marked kindling and glucocorticoids have strong consolidating properties, hence both of these hormones are crucial in development and can alone produce the above syndrome. CRH and glucocorticoids may act in synergy, as in acoustic startle, while glucocorticoids may suppress or stimulate CRH, as in the hypothalamus and amygdala, respectively. A CRH type 1 receptor antagonist, antalarmin, inhibits both the development and expression of conditioned fear in rats, and has anxiolytic properties in monkeys. Profound stressors, such as those from sexual abuse, may elicit the syndrome in older children, adolescents and adults. Most frequently, chronic dysthymia and/or depression may develop in association with gastrointestinal complaints and/or the premenstrual tension syndrome. A lesser proportion of individuals may develop the classic posttraumatic stress disorder, which is characterized by hypocortisolism and intrusive and avoidance symptoms; in younger individuals it may present as dissociative personality disorder. PMID- 12649571 TI - The calculation of target height reconsidered. PMID- 12649572 TI - No influence of surgical stress on postoperative leptin gene expression in different adipose tissues and soluble leptin receptor plasma levels. AB - As part of the postsurgical stress response, plasma leptin levels increase in between 12 h postoperatively. OBJECTIVE: To study the kinetics of leptin gene expression in different adipose tissues before and after severe surgical trauma in children and adults. METHODS: In 22 adults and 23 children with cardiac and 19 adult patients with abdominal surgery, we measured plasma leptin concentrations preoperatively, 4 and 10-17 h postoperatively and quantified leptin mRNA expression by TaqMan real-time PCR in adipose tissue taken at the beginning and the end of surgery from subcutaneous, intrathoracic, omental and mesenteric fat. Plasma-soluble leptin receptor levels were measured in 23 children after cardiosurgery. RESULTS: Plasma leptin levels doubled between 4 and 10-17 h postoperatively in adults (p < 0.001) as well as in children (p = 0.0002) with cardiac surgery. After abdominal surgery, 10-17 h postoperatively, plasma leptin concentrations increased significantly (p < 0.05). During the operation, leptin gene expression did not change in neither of the patient groups. Plasma-soluble leptin receptor levels decreased immediately after the onset of surgery and remained unchanged thereafter. CONCLUSIONS: Leptin gene expression is not up regulated during surgery. The measured increase in plasma leptin after surgery does not result from elevated levels of soluble leptin receptor but may follow an up-regulation of leptin gene expression later after the operation due to postsurgical metabolic changes. PMID- 12649573 TI - Increased abdominal obesity, insulin and glucose levels in nondiabetic subjects with a T29C polymorphism of the transforming growth factor-beta1 gene. AB - BACKGROUND: In humans, a T-->C transition at nucleotide 29 in the region encoding the signal peptide sequence of the transforming growth factor (TGF)-beta(1), which results in a Leu-->Pro substitution at codon 10, has been associated with myocardial infarction. AIMS/METHODS: In the present study, we genotyped 284 unrelated, nondiabetic Swedish men born in 1944 to assess the impact of the Leu10Pro variant on obesity, including abdominal obesity, and estimates of insulin, glucose and lipid metabolism as well as blood pressure. RESULTS: The frequency of the Pro10 variant was 38.9% (95% CI 32.2-46.0%), and the distribution of genotypes was in Hardy-Weinberg equilibrium. Data analysis showed that heterozygotes had significantly higher body mass index compared to homozygous carriers to the Leu10 variant. In addition, homozygous carriers of the Leu10 variant had significantly lower abdominal sagittal diameter than both Leu10Pro and Pro10Pro carriers. We also found that heterozygotes had significantly higher fasting insulin values as well as higher HOMA insulin resistance index in comparison to homozygous carriers of the Leu10. Fasting glucose levels were significantly higher in subjects with the Pro10Pro variant compared to subjects with either the Leu10Leu or Leu10Pro variant. CONCLUSION: These findings suggest that the Pro10 allele in the TGF-beta(1) gene pathway might contribute to prevalent diseases such as obesity and type 2 diabetes mellitus. PMID- 12649574 TI - Increased frequency of islet cell antibodies in unaffected brothers of children with type 1 diabetes. AB - AIM: To assess the relation between islet cell antibody (ICA) positivity and demographic characteristics in an extensive series of first-degree relatives of children with type 1 diabetes (T1D). METHODS: Family members of children diagnosed with T1D before the age of 16 years and attending one of 27 participating paediatric units in Finland taking care of children with diabetes were invited to volunteer for an ICA screening program aimed at identifying individuals eligible for inclusion in the European Nicotinamide Diabetes Intervention Trial (ENDIT). The final series comprised 2,522 first-degree relatives (1,107 males) with a mean age of 20.4 (range 0.1-51.9) years, out of whom 390 were fathers, 622 mothers, 717 brothers, and 793 sisters of affected cases. RESULTS: Two hundred and four family members (8.1%) tested positive for ICA with levels ranging from 3 to 564 (median 18) Juvenile Diabetes Foundation (JDF) units. One hundred and five relatives (4.2%) had an ICA level of 18 JDF units or more. Males had detectable ICA more often than females (9.6 vs. 6.9%; p = 0.02). Antibody-positive family members under the age of 20 years had higher ICA levels than the older ones [median 18 (range 3-514) JDF units vs. 10 (range 3 564) JDF units; p = 0.008]. Among the adult relatives (>or=20 years of age) antibody-positive females had higher ICA levels than the males [median 10 (range 5-564) JDF units vs. 9 (range 3-130) JDF units; p = 0.04]. Siblings had an increased frequency of high-titre ICA (>or=18 JDF units) when compared to the parents (4.8 vs. 3.2%; p = 0.05). Among siblings, we found a higher frequency of ICA positivity in brothers than in sisters (10.8 vs. 6.9%; p = 0.01), and this was also true for high-titre ICA (6.0 vs. 3.8 %; p = 0.04). Geographically, the highest ICA prevalence was seen among relatives living in the middle of Finland (10.4 vs. 7.2% in the other parts of Finland; p = 0.01). CONCLUSIONS: These results imply that male gender and young age favour positive ICA reactivity among family members of children with T1D. Siblings test positive for high ICA titres (>or=18 JDF units) more frequently than parents. Accordingly, judged from demographic characteristics, the yield of ICA screening in first-degree relatives would be maximized by targeting young brothers of affected cases. PMID- 12649575 TI - Hypocalcemic cardiomyopathy due to untreated hypoparathyroidism. AB - Hypocalcemic cardiomyopathy due to hypoparathyroidism is a very rare condition which is usually refractory to conventional treatment for cardiac failure but which responds favorably to restoration of normocalcemia. A 55-year-old man and a 46-year-old woman with a history of postoperative hypoparathyroidism presented with symptoms of cardiac failure and hypocalcemia. A presumptive diagnosis of dilated cardiomyopathy was considered by echocardiography and endomyocardial biopsies were consistent with cardiomyopathy. The coronary angiograms were normal and there was no apparent cause for dilated cardiomyopathy in these patients. The history of the patients and partial recovery of cardiac function after restoration of normocalcemia suggest that hypocalcemia was the cause of dilated cardiomyopathy. PMID- 12649576 TI - 11beta-hydroxysteroid dehydrogenase type 1 deficiency ('apparent cortisone reductase deficiency') in a 6-year-old boy. AB - OBJECTIVE: We present the 1st case of prepubertal hyperandrogenism because of a defect in the conversion of cortisone (E) to cortisol (F) by hepatic 11beta hydroxysteroid dehydrogenase type 1. METHODS AND RESULTS: Clinical and anthropometric data were obtained. Serum androgens and gonadotropins with luteinizing hormone releasing hormone stimulation test, dexamethasone suppression test, and corticotropin-releasing hormone stimulation test were evaluated. Adrenal imaging and urinary steroid profiling by gas chromatography/mass spectrometry were employed. A 6.9-year-old boy presented with precocious pubarche, height (+2.6 SD), accelerated bone age (11.5 years), and Tanner stage 2 pubic hair and genitalia. Serum androgen levels were elevated and dexamethasone suppressible. Serum F was normal, but the E concentration was increased. Central precocious puberty and congenital adrenal hyperplasia were excluded. The excretion of androgen metabolites was moderately increased, but a highly increased tetrahydrocortisone (THE) and a diminished tetrahydrocortisol (THF + allo-THF) excretion was found with a [THF + allo-THF/ THE] ratio of 0.032 (normal controls 1.05 +/- 0.17). The corticotropin-releasing hormone stimulation test showed an exaggerated adrenocorticotropic hormone response, suggesting a relative deficiency of F. Two months of hydrocortisone treatment (17.5 mg daily) failed to suppress androgens adequately. Treatment with dexamethasone (0.375 mg/daily) resulted in androgen suppression. CONCLUSIONS: In the case of precocious pubarche and accelerated growth, the diagnosis of 11beta-hydroxysteroid dehydrogenase type 1 deficiency ('apparent cortisone reductase deficiency') should be considered. The diagnosis is based on determinations of urinary steroid metabolites. PMID- 12649577 TI - [Anti-aging or better aging: a contribution to the prevention of aging?]. AB - Anti-aging wants to influence the quality of the second half of life so that chronic damage to health can also be prevented at a higher age with a good quality of life. Thus, molecular biological tests should become the basis for an individualized, preventive hormone therapy. In this context, the question arises whether for each woman in the menopause a risk profile of disturbances of hormone metabolism can already be recognized in her pattern of polymorphisms, although scientific evidence has been lacking until now. Even the topic of the adrenopause, which--among other features--is characterized by an age-specific, partial adrenal insufficiency, must not be neglected in the context of the problem of anti-aging. PMID- 12649578 TI - [Current concepts in anti-aging]. AB - Anti-aging medicine does not intend to prolong life by all means regardless of the underlying situation. In contrast, anti-aging medicine seeks strategies in order to maintain a high quality of health for people in the second half of their lives, i.e. they should be able to live in their old age without chronic damage to their health. Accordingly, anti-aging strategies have to affect various aspects of life. In Vienna, the following therapeutic concept is currently offered to our patients,which consists of 4 elements: 'lifestyle modification', 'restriction of calories', 'phytotherapy' and our individualized hormone replacement therapy. As estimated by the World Health Organization, the percentage of people aged over 85 will increase dramatically from the year 2010 on. In the light of these data, 'anti-aging medicine' or'preventive medicine' can no longer be neglected. PMID- 12649579 TI - [Hormone replacement therapy, better aging, molecular genetic diagnostics and the image of the gynaecologist]. AB - The benefit of hormone replacement therapy (HRT) for the treatment of menopausal symptoms is well accepted. In contrast, the debate about long-term consequences as a result of continued hormone replacement has recently been reactivated by the publication of results from the Women's Health Initiative Study. The concept of an improved identification of a risk profile of an individual patient based on molecular genetic diagnostics, while highly attractive sofar, has no solid scientific foundation. There is no evidence that by designing HRT on the basis of a specific gene profile of a particular patient the incidence of mild side effects, which are fairly frequent,or of the rare severe complications can be reduced. Innovations in medicine require a convincing benefit-risk evaluation before they become part of routine practice, and a careful attitude of the physician towards new methods in diagnostics and therapy is a fundamental ethical requirement. PMID- 12649581 TI - [Cervical incompetence in multifetal gestation: diagnosis and prophylaxis]. AB - OBJECTIVE: Preterm birth following cervical incompetence threatens infants of multiple gestation. The questions at hand are whether we can validate a sonographic early detection system and if prophylactically intended strategies, such as cervical cerclage, potentially influence pregnancy management and/or perinatal outcome. METHODS: Multifetal pregnancies surveyed with three dimensional ultrasound and pregnancies treated with cervical cerclage were compared to controls. RESULTS: Volumetry of the cervix was possible in all 34 examinations performed. In contrast, two-dimensional cervical length assessment could not be obtained in 6% because the presenting fetal part obstructed the sonographic plane. Mean cervical length was 28.7 mm (SD 7.7). Mean cervical volume was 30.0 cm3 (SD 16.0). A significant correlation was found between mean two-dimensional cervical length and mean cervical volume as both parameters decreased with gestational age (p = 0.01). Prophylactic cervical cerclage was used in 17% of triplet pregnancies studied at a mean gestational age of 16 + 2 weeks (98-138 days). In 50% of the quadruplet/quintuplet pregnancies studied, the cerclage was performed at a mean gestational age of 15 + 2 weeks of gestation (78 152 days). The time interval from operation to delivery was 106 days (62-119) for triplets and 96 days (57-142) for quadruplets/quintuplets. Prophylactic cervical cerclage did not prolong pregnancies compared to controls. With respect to the need for hospitalization or intravenous tocolysis or perinatal outcome parameters, no benefit was achieved. CONCLUSIONS: The results disclaim a positive impact of prophylactic cervical cerclage on the course of a multifetal pregnancy and/or perinatal outcome. On the other hand, early non-invasive diagnosis of cervical incompetence enables a risk-adapted conservative pregnancy management. PMID- 12649580 TI - [Adrenopause and dehydroepiandrosterone: pharmacological therapy versus replacement therapy]. AB - BACKGROUND: Adrenopause is an age-related, partial insufficiency of the adrenal cortex characterized by its low blood levels of dehydro-epiandrosterone (DHEA) and DHEA sulfate (DS) in the presence of undiminished cortisol levels. A great number of effects in the CNS and in the periphery are known, partially due to DHEA as an independent hormone, partially due to its influence as a precursor of sex hormones. Positive epidemiological data about DHEA on morbidity and mortality in males as well as clinical reports about promising effects of a DHEA supplementation in elderly people suggest that controlled replacement therapy might be useful in the prevention and treatment of degenerative processes in humans. In studies sofar a fixed daily DHEA dose of 50-100 mg in men and 25-50 mg in women was used irrespective of the individual extent of the adreno-pause. This regimen raised serum levels of DS to and mostly above the upper normal range, thus leading to pharmacological effects. PATIENTS AND METHODS: We report about our experiences in DHEA dose finding in 100 men and 100 women with adrenopause, aged between 46 and 74 years, over a period of 6-12 weeks in order to find a suitable daily oral DHEA dose. The aim was to raise the daily DS peak level between 3 and 5 h after the intake in the morning to 2.0-2.8 pg/ml in women and 4.0-5.0 gg/ml in males, levels that are thought to be in the optimal range of healthy adults in the third decennium of their life (controls). RESULTS: There were 5, 10, 15, 25 or 50 mg DHEA in 18, 26, 34, 19 or 3% of the women and 15, 25, 50, 75 or 100 mg DHEA in 5, 13, 51, 17 or 14% of the males suitable for that purpose. This adjusted dose regimen raised serum levels (mean values +/- SD) significantly (p < 0.01): (a) in women in the case of DS from 0.7 +/- 0.4 to 2.4 + 0.5 microg/ml, free testosterone from 0.4 +/- 0.4 to 0.9 +/- 0.5 pg/ml and androstenedione from 0.8 +/- 0.4 to 1.4 +0.4 ng/ml, and (b) in males in the case of DS from 1.4 +/- 0.5 to 4.1 +0.7 microg/ml, free testosterone from 10.9 + 4.1 to 14.7 +/- 4.5 pg/ml,androstenedione from 1.2 +/- 0.5 to 2.0 +/- 0.6 ng/ml, estrone from 28 +/- 14 to 41 +/- 19 pg/ml and estradiol from 16 +/- 8 to 31 +/- 15 pg/ml. In cases of inadequate dosage there were side effects like sleepiness,restlessness, headache, acne/hirsutism, effluvium or odors in a percentage of 34, 17, 9, 31, 21 and 11% of the women, respectively.After having adjusted the individual dosage to meet the proper serum levels of DS, these side effects were significantly reduced (p <0.05; p < 0.2 in cases with headache) and found only in 8, 2, 4, 6, 7 and 0%, respectively. In males, such symptoms occurred only occasionally. CONCLUSION: We suggest replacement therapy in cases of adrenopause with an "individually adjusted" low DHEA dose between 5 and 50 mg for women and 15 and 100 mg for men in order to raise DS peak levels into the physiological range of younger adults. This procedure has here been applied routinely for the last 5 years, leading to an excellent compliance of the patients. In contrast,a high-dose pharmacological DHEA administration seems to be suit-able for patients with systemic lupus erythematosus and other related diseases. PMID- 12649582 TI - [Sentinel lymph node biopsy in breast cancer patients--results and experience after 500 sentinel lymph node biopsies]. AB - INTRODUCTION: Sentinel lymph node biopsy (SLNB) is a widely used technique for axillary staging in breast cancer patients. The principle to evaluate the axillary status of a breast cancer patient with a less invasive surgery than axillary lymph node dissection (ALND) meets the new minimally invasive concept in breast cancer surgery. Some breast cancer centers proceed to SLNB without ALND in SLN-negative patients. PATIENTS AND METHODS: Between March 1998 and March 2002, 500 SLNBs were performed. After a learning period with SLNB and ALND in 75 patients with a sensitivity of 96.2% and a false-negative rate of 3.8%, SLNB alone without further ALND was performed in a group of patients. In addition, the feasibility of SLNBin patients with locally advanced breast cancer, in patients after neoadjuvant chemotherapy and in patients with multicentricity was evaluated. The combined method with blue dye and technetium-99m-labeled human albumin for identification of SLNs was applied. RESULTS: 500 SLNBs were performed. The identification rate was 86.2%. After exclusion of patients with neoadjuvant chemotherapy and patients with multicentricity, the identification rate was 94.5%. SLNs were positive in 41.3% of patients and negative in 58.7% of patients. DISCUSSION: SLNB is an excellent method for axillary stag-ing and an alternative for ALND in a certain group of breast cancer patients. PMID- 12649583 TI - [Multi-institutional survey on the value of lymphadenectomy in endometrial carcinoma in Germany]. AB - INTRODUCTION AND OBJECTIVE: Primary surgical staging has been established in the management of carcinoma of the endometrium. Various working groups have also recommended the use of lymphadenectomy. Contrary to the prognostic relevance of lymph node status, the therapeutic value of lymphadenectomy is still unclear. METHOD: A total of 1,030 standardised questionnaires were sent to gynaecological clinics (including in-patient clinics) in Germany as part of a cross-sectional study between October 2001 and January 2002. RESULTS: It was possible to evaluate 595 (58%) of the questionnaires from 5.2% university clinics, 69.7% clinics for maximal treatment and 25% clinics for regular therapy. Is a lymphadenectomy performed in the case of a carcinoma of the endometrium? Only 7.3%(95% Cl: 5.3 9.7%) reported that they never performed a lymphadenectomy, 62.9% (95% Cl: 58.9 66.9%) performed one in special cases, 29.7 (95% Cl: 26-33.7%) did so regularly. Which methods do you use for lymphadenectomy? 27.1% (95% Cl: 23.2-31.4%) used'sampling' and 72.9% (95% Cl 68.6-76.8%) used the technique of systematic removal. 26.6% lymph node dissections were both pelvic as well as para-aortic. What is your intention when performing a lymphadenectomy? 31.7% (95 Cl: 22.8 30.6%) performed it for stag-ing reasons, 3.1% (95% Cl: 1.8-4.9%) for the patient's survival and 65.2% (95% Cl: 60.9-69.3%) for both of these reasons. 94.5% (95% Cl:92.1-96.3%) said that the status of the affected lymph nodes influenced the choice of adjuvant treatment. 76.8% (95% Cl: 72.8-80.4%) and 36.9% (95% Cl: 46.9-56.5%) chose radiotherapy in stages I/II and III/IV, respectively; 20.4% (95% Cl: 16.9-24.2%) and 51.7%(95% Cl: 46.9-56.5%) chose radiochemotherapy. CONCLUSIONS: The conception of prospective, multicentric studies of operative and adjuvant management should be based on these results. PMID- 12649584 TI - [Is continuing medical education under suspicion of corruption? Contribution to the discussion by the Committee for Quality Preservation of the Swiss Society of Gynaecology and Obstetrics]. AB - The requirements laid down by law and by the medical profession itself with regard to continuing medical education have recently been tightened. The cost of good training is high, both in time and money. Sponsoring, especially by the pharmaceutical industry, has helped to keep costs down for individual participants. The question of a tariff system for the payment of compulsory training is now being raised. Rates have been set to ensure that, in the future, indirect costs are covered. In the event of a full tariff-based payment of costs being introduced, various disadvantages could be expected both for participants and the quality of the training. Changes in Swiss criminal law and the new medicines act do not forbid sponsoring, but they do call for rules of professional ethics and of personnel law in public hospitals governing behaviour in respect of funding by sponsors. Various sponsoring models are conceivable with differences in allocation to organizers, individual participants or distribution via a pool system. There are strong arguments in favour of continuing the existing practice of mixed funding by participants and sponsoring, provided that it is subject to certain rules of transparency. What is lacking to date is a political decision on the issue of funding for continuing medical education. PMID- 12649585 TI - Embryonic development of the pituitary gland in the chick. AB - Pituitary glands of chicken, from stages 20 (70 approximately 72 h of incubation) to 46 (20 days) of Hamburger and Hamilton (1951), were studied by immunocytochemical and histological stainings and India ink injection into blood vessels. Using the distribution pattern of 6 types of immunoreactive adenohypophyseal cells and the location of pituitary stalk as guideposts, we found how specific areas in the epithelium of Rathke's pouch differentiate into specific regions of the adenohypophysis at 20 days. In the sagittal plane, the walls of Rathke's pouch were tentatively divided into the upper part (A(1) + A(2)) and lower part (A(3)) of the anterior wall, and the posterior wall (P(1) + P(2) + P(3)). The cephalic lobe was mainly assembled by the proliferation of parenchymal cells in the areas A(2) + A(3) + P(2) of Rathke's pouch epithelia at 3 days of incubation. The caudal lobe was derived from A(1) + P(1) + P(3). The pars tuberalis was derived from A(1) + A(2). Thus, the avian adenohypophysis is established at 13 days, though the blood supply to the pars distalis is established at 20 days. Therefore, the cephalic lobe and caudal lobe of the pars distalis and the pars tuberalis of the chicken adenohypophysis are derived from specific areas of the cell cords of Rathke's pouch at 3 days of incubation. PMID- 12649586 TI - Somites, spinal Ganglia, and centra. Enumeration and interrelationships in staged human embryos, and implications for neural tube defects. AB - Serial sections of 99 human embryos from Carnegie stages 8-23 were investigated and 38 graphic reconstructions were evaluated. At stage 9 somite 1 is of appreciable size and is separated from the otic disc, as also in the next several stages by rhombomeres and pharyngeal arches 3 and 4, thereby differing from the chick. At stage 10 somite 1 begins to differentiate into sclerotome and dermatomyotome. At stage 11 spinal neural crest begins to develop. At stage 12 parts of somites 1-4 are being transformed into the hypoglossal cell cord. It is stressed that the numbers of somites present at stages 9-12 are part of the definition of those stages. At stage 13 dense and loose zones begin to be detectable rostrally in the sclerotomes and also, although out of phase, in the perinotochord. Spinal ganglia begin to develop in phase with the somites. At stages 14-16 the maximum number of somites observed was 38-39 rather than 42-44, as usually given. Moreover, they did not extend to the tapered end of the trunk, which is not a (vertebrated) 'tail'. At stages 17-23 the maximum number of centra was 38-39, including coccygeal vertebrae 4-5. Although most of the somites appear during primary development, all of the spinal ganglia develop during secondary development (stages 13-18). The number of ganglia was at a maximum of 35 at stage 18, but was reduced to 32 already by stage 23. Important points confirmed in this study are that the number of occipital somites in the human is four, and that the level of final closure of the caudal neuropore is future somite 31, which represents approximately future sacral vertebra 2. The interpretation of relevant neural tube defects is discussed in the light of the findings. The ascensus of the conus medullaris during the fetal period is well established, but a concomitant ascent of the situs neuroporicus is proposed here, and has implications for defects that involve tethering of the spinal cord. The main results are integrated in comprehensive graphic representations of the levels and the interrelationships of (a) somites and centra, and (b) somites, neural crest, and spinal ganglia. These may aid in the elucidation of some frequently occurring anomalous conditions. PMID- 12649587 TI - Hyaluronan synthases in normal and regenerating joint cartilage. AB - Repair of full thickness joint cartilage defects is within reach of routine clinical practice. The quality of regenerating hyaline cartilage, however, is difficult to assess. Synthesis of an extracellular matrix with high hyaluronan content is crucial for its metabolic and functional properties. We studied hyaluronan synthase (HAS) expression in knee joints of adult sheep as a novel cellular marker for chondrocyte function. Six house-bred Merino sheep (age 4-6 years) underwent two-stage surgery of their femoral condyles for autologous chondrocyte transplantation (ACT). First, cells were isolated from biopsies and expanded in vitro for 2 weeks using standard culture techniques. In a second session, three defects were made and either left untreated, covered with periosteal flap alone, or in combination with chondrocyte suspensions injected under the flaps. After 16 weeks, biopsies were taken from the operated knees at the defect sites and from the untreated condyle. Specimens were processed for safranin O and electron microscopy, and for immunofluorescence using three different polyclonal anti-HAS antibodies recognizing one or all of the three known mammalian HAS. Control and regenerating tissues were compared regarding their morphology and the expression of HAS, in relation to collagens type I and II, and adult cartilage proteoglycan core protein. In comparison with untreated defects or with periosteal flap alone, ACT provided a neocartilage with better differentiated morphology. In healthy joint cartilage, about 50% of the chondrocytes expressed HAS, independent of antibody. Following ACT, a higher density of chondrocytes was observed, of which more than 75% expressed HAS, whereas the regenerates without ACT showed a lower density of HAS-expressing cells. We propose to use HAS immunofluorescence as an additional marker of matrix synthesis by chondrocytes and joint cartilage regeneration. PMID- 12649588 TI - Morphological characteristics of the interpubic joint (Symphysis pubica) of rats, guinea pigs and mice in different physiological situations. A comparative study. AB - The pubic joint of male and female rats, guinea pigs and mice was studied using the Picrosirius polarization method which selectively discloses the fibers of the collagenous system. Besides that, considerations were made regarding joint classification. In adult rats (both males and females, including intrapartum specimens), our results confirm those of earlier studies showing that the interpubic joint contains a central core of hyaline cartilage surrounded by fibrocartilaginous areas. Thus, in rats, the pubic joint should more properly be classified as a true synchondrosis. In virgin female guinea pigs and mice, the interpubic joint is formed of fibrocartilage (a true symphysis); whereas at term the bones are joined by a connective ligament, constituting a syndesmosis. Male mice have a similar (fibrocartilaginous) joint structure to virgin female mice, whereas male guinea pigs (like rats) have a hyaline cartilage joint. The foregoing observations indicate that the classification of the pubic joint depends upon the species, age, sex, and physiological reproductive stage studied. Species that are very similar in most other aspects (such as rats and mice) displayed different morphological features of the pubic joint to support the same reproductive processes. Together, the data reported here suggest that interspecies differences are likely to be found in other parameters and should be considered when choosing an appropriate animal model for research or teaching purposes. PMID- 12649589 TI - A histological investigation of the lymphoid and immunohaematopoietic tissues of the adult stripe-faced dunnart (Sminthopsis macroura). AB - This is the first published description of the lymphoid and immunohaematopoietic tissues of an Australian polyprotodont, the stripe-faced dunnart, Sminthopsis macroura and the first account of bronchus-associated lymphoid tissue (BALT) in a metatherian. Histologically, the tissue beds are similar in appearance to those reported in other adult eutherian and metatherian mammals. The liver and bone marrow were mature and virtually no haematopoietic activity was observed. The thymus had undergone involution but retained some lymphocytes. The spleen was similar to that observed in other metatherians containing areas of red and white pulp separated by a marginal zone. Lymph nodes, except for a pair in the posterior abdomen, were difficult to locate but were similar to those observed previously in other adult metatherians. Peyer's patches were present; however, they lacked dome regions and sometimes had villi above them. BALT appeared to be both compartmentalised and non-compartmentalised in the adult stripe-faced dunnart. PMID- 12649590 TI - Mesoscopy and scanning electron microscopy of the trabecular projections in the superior sagittal sinus. AB - The trabecular projections of the human superior sagittal sinus were classified into types and subtypes according to spatial arrangement and shape. The horizontal and vertical projections direct laminar blood flow, while the conic type, which is avalvular, protects the openings of the superior cerebral veins in the superior sagittal sinus. PMID- 12649591 TI - Electrophysiological characterization of cardiac muscarinic acetylcholine receptors: different subtypes mediate different potassium currents. AB - To characterize electrophysiologically the K+ currents mediated by various mAChR subtypes, we performed detailed whole-cell patch-clamp studies in canine atrial myocytes. I(KACh) was induced by 1 mM ACh (acetylcholine) or by arecaidine but-2 ynyl ester tosylate (100 nM, an M2 receptor selective agonist) and was blocked by methoctramine (20 nM, an M2 receptor selective antagonist). Tetramethylammonium (0.5 mM) activated a K+ conductance with delayed rectifying properties (I(KM3)) and the currents were highly sensitive to 4-diphenylacetoxy-N-methylpiperidine methiodide (2 nM, an M3 receptor inhibitor). 4-aminopyridine (1 mM) induced a delayed rectifier-like current (I(K4AP)) which was selectively suppressed by tropicamide (200 nM, an M4 receptor blocker). The current waveforms, I-V relationships, steady-state voltage-dependence, kinetics and pharmacological properties of these three currents were different from one another and distinct from the classical delayed rectifier K+ currents (I(Kr) and I(Ks)). Both I(KACh) and I(K4AP) were sensitive to pertussis ntoxin, whereas I(KM3) was not. Isoproterenol (1 mM) markedly depressed I(KM3), but increased I(K4AP) and did not alter I(KACh). The effects of isoproterenol were reversed by propranolol (1 mM); and ACh completely suppressed I(KM3) and I(K4AP). The results suggest that the K+ currents mediated by different subtypes of mAChR represent different populations of K+ channels and that the cholinergic regulation of the heart's electrical function is a consequence of activating multiple mAChRs linked to different effector systems with potentially varying signal transduction. PMID- 12649592 TI - Fluid shear stress differentially regulates gpr3, gpr6, and gpr12 expression in human umbilical vein endothelial cells. AB - Fluid shear stress is a major factor involved in the control of gene expression in vascular endothelial cells. Sphingosine 1-phosphate has emerged as a multifaceted regulator of endothelial cell function and its high affinity S1P1 receptor is one among the many shear stress regulated genes. We recently identified the orphan G protein coupled receptors gpr3, gpr6 and gpr12 as additional S1P receptors. Here, we investigated their expression in various human endothelial and vascular smooth muscle cell lines via RT-PCR and western blot analysis. We next sought to determine the role of fluid shear stress in the regulation of expression of gpr3, gpr6 and gpr12 by using human umbilical vein endothelial cells (HUVECs). Laminar shear stress (12dyne/cm2) did not significantly increase gpr3, gpr6 and gpr12 mRNA after 1, 2, 4, 6, and 8 hrs of application of elevated pressure as determined by quantitative Taqman RT-PCR analysis. In contrast, gpr3 and gpr12 protein were increased after 12 hrs of shear stress by 95% and 40%, respectively. gpr6 mRNA and protein were absent in HUVECs as determined by Taqman and western blot techniques. Our results suggest that shear stress regulates gpr3 and gpr12 but not gpr6 expression and that regulation does not occur transcriptionally but posttranslationally. gpr3 and gpr12 may therefore add to the repertoire of S1P receptors, translating extracellular S1P effects into intracellular signals in human endothelial cells. PMID- 12649593 TI - STAT5a regulates the GlcNAc-1-phosphate transferase gene transcription and expression. AB - The dolichyl-phosphate alpha-N-acetylglucosaminephosphotransferase 2 (Dpagt2) gene in the mouse has a housekeeping promoter, and its expression is regulated during the development and hormonally modulated lactogenesis of the mammary gland. Previous studies showed that the transcription of the mouse mammary Dpagt2 gene is stimulated by the lactogenic hormones, insulin, glucocorticoid receptor (GR), and prolactin. Transcription factors which bind to the Dpagt2 gene promoter region can influence the expression level of the Dpagt2 gene. It is supposed that the Dpagt2 gene promoter region (bases pairs -1462 to -5) maybe contain 10 putative STAT (signal transducer and activator of transcription) binding sites: TTN (5/6) AA. In order to identify the STAT factors involved in the transcription of the GPT gene, 32P labeling probes and lactating mouse nuclear extracts were prepared. Electrophoretic Mobility Shift Assays (EMSA) show that the region (bases pairs -386 to -322, where there is a STAT binding site, TTTCAAAAA) binds to STAT5a, not to glucocorticoid receptor (GR) or other STAT factors. The involvement of STAT5a in regulating the expression of the mouse Dpagt2 gene was further investigated by transient transfections of various Dpagt2 promoter/luciferase (Luc) constructs into COS 7 cells. The results showed that co transfection of STAT5a or prolactin receptor can enhance Dpagt2 promoter activities in the promoter construct pGL-MX6 (from base pairs -386 to -5), but not in the promoter construct pGL-MX7 (from base pairs -322 to -5). This paper first reports that STAT5a is involved in the binding between -386 and -322 base pairs of the Dpagt2 gene promoter and stimulates the expression of the Dpagt2 gene transcription in the mouse lactating mammary gland. PMID- 12649594 TI - Effect of ATP on intracellular pH in pancreatic ducts involves P2X7 receptors. AB - Pancreatic acini release ATP, which can stimulate HCO3--secreting ducts that express purinergic receptors from both P2X and P2Y families. The aim of this study was to investigate whether extracellular ATP affects HCO3- or H+ transport across the plasma membrane of intralobular ducts, and determine which P2 receptors might be involved. Ducts were obtained from rat pancreas, and the pH sensitive fluorophore BCECF was used to measure pHi and recovery rates from cellular acidosis induced by ammonium pre-pulses. In order to reveal Na+/H+ exchange, Cl-/HCO3- exchange or a Na+-HCO3- cotransport, experiments were performed in solutions with or without bicarbonate buffers (+BIC or -BIC), with amiloride derivative EIPA, or with low extracellular Cl- concentrations. Although these transporters contributed to pHi recovery from acidosis, ATP had no effect. Nevertheless, ATP induced a small and reversible decrease in pHi by 0.07+/-0.02 pH-units and BzATP decreased pHi by 0.29+/-0.07 pH-units in -BIC (n=10, 11). These effects were abolished by Brilliant Blue and in Ca2+-free solutions. Our study shows that the pHi effect of ATP is mediated by P2X7 receptors. However, ATP does not affect H+/HCO3- transporters unmasked by cellular acidosis. Presumably, ATP alone does not stimulate HCO3- secretion in pancreatic ducts. PMID- 12649595 TI - Cryopreservation of rat precision-cut liver slices is associated with major metabolic stress and ionic perturbations. AB - BACKGROUND/AIMS: Cryopreserved precision-cut liver slices (PCLS) in culture exhibit a rapid decline in ATP level and anabolic processes by unknown mechanisms. The aim of this study is to elucidate the key events explaining the alterations occurring in cryopreserved PCLS. METHODS: Glucose metabolism, mitochondrial activities, ionic homeostasis and caspase-3 activity were assessed in fresh or cryopreserved (rapid freezing-thawing conditions) rat PCLS within the first hour of incubation in Williams' medium E at 37 degrees C. RESULTS: Despite a similar glycolytic activity under both conditions, only fresh PCLS were able to gradually recover their ATP and potassium content. Glycogen content dropped more rapidly in cryopreserved than in control PCLS. Only cryopreserved PCLS exhibited a decline in O2 consumption and a lower ATP/ADP ratio from 30 min of incubation with a loss of coupling of oxygen consumption to ATP synthesis and mitochondrial calcium transport after 1 hour of incubation. Caspase-3 activation was already present in cryopreserved PCLS after a few minutes of incubation. CONCLUSIONS: The lack of restoration of potassium/sodium exchange might be a primary event in the metabolic alterations occurring in cryopreserved PCLS after thawing, that contributes to further mitochondrial alterations, caspase activation and compromises ATP-dependent anabolic pathways. PMID- 12649596 TI - Cell volume induced hormone secretion: studies on signal transduction and specificity. AB - Cell swelling causes an immediate secretory response in various cell types. Induced secretion possesses some unique features suggesting the involvement of a specific signal transduction pathway. The effect of 10-20 microM GdCl3, 100 microM HgCl2, 1-100 microM indomethacin and 1-20 microM nordihydroguaiaretic acid (NDGA) on cell swelling-induced hormone secretion (isosmotic 80 mM ethanol or 15 30% hyposmotic medium) from incubated rat hypothalamic paraventricular nucleus (PVN) and posterior pituitary (oxytocin and TRH), isolated pancreatic islets (TRH) and perifused anterior pituitary cells (prolactin) were examined. To determine how general the effect of cell swelling is on exocytotic secretion, the release of two different neurohormones (thyrotropin releasing hormone -TRH and oxytocin) from the same tissue explant were studied. Both hyposmotic medium or isosmotic ethanol containing medium induced immediate TRH and prolactin release from the tested tissues. The effect of GdCl3, HgCl2, NDGA or indomethacin showed no inhibition of cell swelling induced secretion. In contrast to TRH, oxytocin release was not induced by isosmotic ethanol containing medium from the PVN or posterior pituitary. CONCLUSION: These data indicate that signal transduction leading to exocytosis after cell swelling does not involve GdCl3 sensitive stretch activated channels, mercury sensitive aquaporins, or indomethacin and NDGA sensitive mediators including prostaglandins and leukotriens. Cell swelling induced exocytosis possesses limited selectivity; cells specifically involved in water and salt regulation retain their specific response to osmotic stimuli. PMID- 12649597 TI - Stimulus-dependent regulation of serum and glucocorticoid inducible protein kinase (SGK) transcription, subcellular localization and enzymatic activity. AB - We originally discovered the serum and glucocorticoid inducible protein kinase, SGK, as a novel protein kinase that is under acute transcriptional control by serum and glucocorticoids. An expanding set of cell surface receptor, nuclear receptor, and cellular stress pathways has been shown to target SGK, which has implicated this regulated signaling molecule in a variety of biological functions. Compared to most other protein kinases, a distinguishing feature of SGK is the stringent stimulus-dependent regulation of its transcription, subcellular localization and enzymatic activity. In addition, SGK expression is regulated during discrete developmental stages, and during normal and abnormal physiological function. An analysis of the SGK promoter reveals many potential transcription factor sites that potentially account for the stimulus-dependent changes in SGK transcript expression observed in a variety of cell systems, although, the direct stimulus regulation of SGK promoter activity has been established only for glucocorticoids, p53 tumor suppressor protein, hyperosmotic stress and follicle stimulating hormone. In the systems tested to date, hormones, growth factors and environmental cues induce expression of a catalytically active SGK. It is now well established that the enzymatic activity of SGK is controlled by the PI 3-kinase cascade which produces a hyperphosphorylated active SGK. A critical third level of regulation is the stimulus-dependent control of SGK subcellular localization. The nuclear-cytoplasmic shuttling of SGK is regulated by a nuclear localization signal (NLS) that binds to the importin-alpha nuclear import receptor. Modeling of the 3-D structure of the central region of SGK that includes the kinase domain predicts that the NLS is located at an external surface of the molecule. Thus, multiple signal transduction pathways converge on SGK to control its availability, function and access to its substrates and non substrate targets. PMID- 12649598 TI - SGK1 regulation of epithelial sodium transport. AB - Epithelial ion transport is regulated in vertebrates by a variety of hormonal and non-hormonal factors, including mineralocorticoids, insulin, and osmotic shock. SGK1 has been established as an important convergence point for multiple regulators of Na+transport. Unlike most serine-threonine kinases, SGK1 is under dual control: protein levels are controlled through effects on its gene transcription, while its activity is dependent on phosphatidylinositol-3-kinase (PI3K) activity. Aldosterone is the most notable regulator of SGK1 protein level in ion transporting epithelia, while insulin and other activators of the of PI3K are key regulators of its activity. Activated SGK1 regulates a variety of ion transporters, the best characterized of which is the epithelial sodium channel (ENaC). The apical targeting of ENaC is controlled by the ubiquitin ligase, Nedd4 2, and SGK1 acts, at least in part, through phosphorylation-dependent inhibition of Nedd4-2. This effect of SGK1 requires physical associations of Nedd4-2 with both SGK1 and ENaC. Moreover, direct physical association between SGK1 and ENaC may also be implicated in the formation of a tertiary complex. Osmotic shock is likely the most important non-hormonal regulator of SGK1 expression, and surprisingly, SGK1 expression can be induced by hypotonic or hypertonic stress in a cell-type dependent fashion. The SGK family represents an ancient arm of the serine-threonine kinase family, present in all eukaryotes that have been examined, including yeast. SGK1 appears to have been implicated in membrane trafficking and possibly in the control of ion transport and cell volume in early single cell eukaryotes. In metazoan epithelia, it seems likely that SGK1 was adapted to the regulation of ion transport in response to hormonal and osmotic signals. PMID- 12649599 TI - SGK1: aldosterone-induced relay of Na+ transport regulation in distal kidney nephron cells. AB - Aldosterone increases within 30 min renal Na+reabsorption and K+secretion by a mechanism that is triggered at the level of gene transcription. Thus, gene products that are rapidly up- or down-regulated transmit this effect to the transport machinery within the distal nephron target cells. One such rapidly up regulated gene product is a structural element of the transport machinery, namely the a subunit of ENaC. Its amount might in certain conditions play a rate limiting role for Na+transport. Cell-surface localization and function of ENaC and of the Na,K-ATPase are also tightly controlled by a complex regulatory network and aldosterone appears to acutely regulate the expression of elements of this network such as the small G-protein K-Ras (in A6 cells) and the kinase SGK1 (also in ENaC-expressing cells of the mammalian distal nephron). The kinase SGK1 is an early aldosterone-induced protein that relays signals from pathways that are transmitted via PDK1/2 and possibly PKA. Active SGK1 has been shown to increase ENaC and Na,K-ATPase cell-surface expression in Xenopus oocytes. This effect at the level of ENaC has been recently shown to be mediated by the ubiquitin ligase Nedd4-2 which is a direct target of SGK1. Once phosphorylated by SGK1, Nedd4-2 is prevented from interacting with ENaC and thus from decreasing ENaC cell-surface expression. This SGK1-Nedd4-2-ENaC pathway is the first direct link between aldosterone-induced transcriptional regulation and the function of the Na+transport machinery to be unravelled. The physiological importance of this pathway for mediating the aldosterone response in different target epithelia remains to be verified in vivo, in particular in view of the axial gradient of ENaC apical translocation observed along the aldosterone-sensitive distal nephron. PMID- 12649601 TI - Regulation of channels by the serum and glucocorticoid-inducible kinase - implications for transport, excitability and cell proliferation. AB - The serum and glucocorticoid-inducible kinase SGK1 stimulates the Na+ channels ENaC and SCN5A, the K+ channels ROMK1, Kv1.3, and KCNE1/KCNQ1, the cation conductance induced by 4F2/LAT1 and the chloride conductance induced by CFTR. The isoforms SGK2 and SGK3 have similarly been shown to regulate ENaC, SCN5A, Kv1.3 and KCNE1/KCNQ1. The kinases regulate channel abundance in the plasma membrane in part by inhibition of the ubiquitin ligase Nedd4-2 and in part by interaction with trafficking molecules such as the Na+/H+ exchanger regulating factor NHERF2. An in vivo role of SGK1 mediated ENaC channel regulation in renal salt excretion and blood pressure control is documented by the impaired ability of SGK1 knockout mice to adequately reduce renal Na+ output and maintain blood pressure during dietary salt restriction and by enhanced blood pressure in individuals carrying certain polymorphisms in the SGK1 gene. The in vivo physiological significance of SGK dependent regulation of the other channels remains to be shown even though circumstantial evidence points to involvement in the regulation of epithelial transport, cell volume, cell proliferation, cardiac action potential and neuroexcitability. There is little doubt that further channels will be identified which are modulated by the SGKs and that further in vivo physiological functions will be defined where channel regulation by the SGKs plays a critical role. PMID- 12649602 TI - Twin studies in the analysis of minor physiological differences between individuals. AB - To test the hypothesis that the serum and glucocorticoid regulated kinase (SGK1) is of relevance to blood pressure in man, we recruited monozygotic (MZ) (126 pairs) and dizygotic (DZ) (70 pairs) normotensive twin subjects and parents of DZ twins. Blood pressure was measured in a controlled fashion recumbent, sitting, and upright. We documented genetic variance on blood pressure in all positions. We then relied on microsatellite markers at the SGK1 gene locus (D6S472, D6S 1038, and D6S270) and two single nucleotide polymorphisms within the SGK1 gene. We found significant linkage of the SGK1 gene locus to diastolic blood pressure (p<0.0002) and suggestive evidence for linkage for systolic blood pressure (p<0.04), documenting the locus as a QTL for blood pressure. We next performed association, using all DZ twins and an MZ member from each pair. We found significant associations between both SNP variants and blood pressure, as well as a significant interaction between the SNPs enhancing the effect. This combined effect of the polymorphisms was confirmed in an independent sample of 260 young normotensive men. These data, coupled with our earlier observations linking the insulin-like growth factor-1 gene locus to blood pressure lead us to conclude that the SGK1 gene is relevant to blood pressure regulation and probably to hypertension in man. PMID- 12649603 TI - Contents of the Cochrane library on the organisation of stroke services. AB - BACKGROUND: The Cochrane Collaboration is an international organisation that aims to help people make well-informed decisions about healthcare by preparing, maintaining and promoting the accessibility of systematic reviews of the effects of healthcare interventions. This paper briefly summarises the output of the Cochrane Stroke Group in relation to evaluating different forms of the organisation of services for patients with stroke. METHODS: A survey of randomised controlled trials and systematic reviews (and meta-analyses) of randomised controlled trials. RESULTS: Issue 2 of the Cochrane Library 2002 includes 1,377 completed systematic reviews and 1,055 protocols for reviews in progress. The Cochrane Controlled Trials Register included 343,270 reports of randomised or controlled clinical trials. The Cochrane Stroke Review Group had contributed 63 completed systematic reviews, 29 protocols of reviews in progress, and 4,396 reports of stroke trials. The main published reviews relevant to entire services related to: stroke units; services for helping acute stroke patients avoid hospital admission; services for reducing duration of hospital care for acute stroke patients; care pathways for stroke. MAIN CONCLUSIONS: There has been a rapid recent growth in the evidence base on stroke care. Systematic reviews of several aspects of stroke services are now published and other reviews are in progress. For areas where no reviews have been completed, the Cochrane Library includes a substantial number of individual randomised trials evaluating the effects of specific services and components of services. PMID- 12649605 TI - Stroke units and stroke services in Germany. AB - BACKGROUND/OBJECTIVES: The German stroke units are sub-intensive units with the possibility of continuous monitoring of vital parameters. This is the main difference to Scandinavian and British non-intensive combined acute and rehabilitation stroke units. Germany has 56 regional and 50 local stroke units, and standards differ between them. Nearly 30% of all strokes in Germany are treated in these units. The German stroke units are cost intensive and unfortunately their effectiveness has not been proven yet. PMID- 12649604 TI - Riks-stroke - a Swedish national quality register for stroke care. AB - BACKGROUND: Riks-Stroke, the Swedish national quality register on stroke care, provides unique opportunities to evaluate stroke units in routine clinical care. METHODS: Basic patient characteristics, process indicators and outcome variables are recorded in all 85 hospitals admitting acute stroke patients. A 3-month follow-up is included. RESULTS: There are wide variations between hospitals in the proportion of patients admitted to a stroke unit, in secondary prevention and in the proportion of patients in institutional care at 3 months. Even after adjustment for available prognostic indicators, case fatality is lower and functional outcome is better in patients treated in stroke units than in patients treated in general wards. CONCLUSION: Riks-Stroke shows that outcome is consistently better in patients treated in a stroke unit than in general wards, not only in randomised trials but also in routine stroke care. PMID- 12649600 TI - Concerted roles of SGK1 and the Na+/H+ exchanger regulatory factor 2 (NHERF2) in regulation of NHE3. AB - Na+/H+ exchanger regulatory factors, NHERF1 and NHERF2, are structurally related proteins and highly expressed in epithelial cells. These proteins are initially identified as accessory proteins in the regulation of Na+/H+ exchanger isoform 3, NHE3. In addition to regulation of NHE3, recent studies demonstrate the importance of NHERF1 and NHERF2 in recycling and localization of membrane receptors, ion channels and transporters. Recent studies show that serum- and glucocorticoid-induced kinase 1 (SGK1) specifically interacts with NHERF2 but not with NHERF1, adding to the growing number of differences between the two proteins. The association of SGK1 with NHERF2 is necessary for stimulation of NHE3 activity by glucocorticoids. In addition, SGK1 together with NHERF2 stimulates the K+ channel ROMK1, suggesting a broader role of SGK1 in regulation of ion transport. PMID- 12649606 TI - The French stroke program. AB - The French healthcare system is organised according to a political, geographical and demographic framework. In France today, less than 20 hospitals have dedicated intensive care units for acute stroke patients. In the 1999 national survey, 94% of stroke patients were admitted to public hospitals, 35% were treated in neurological wards and 4% had access to an intensive care stroke unit. To improve the quality of care and resource utilisation, a national plan for stroke will be implemented at regional level during the next five-year plan. After a long period of status quo, many positive actions for stroke care have been initiated within a couple of years, based on the evidence of more effective stroke treatments, such as thrombolysis and the specialist care provided by stroke units. PMID- 12649607 TI - The polish experience in early stroke care. AB - Stroke units are being organised in Poland since 1997. To ensure easy access to stroke care, 120 specialised units need to be created. There were 23 stroke units in Poland in the year 2000. Only 12 of them fulfilled the criteria of a standard stroke unit. Significant differences in stroke fatality were noted between well- and as yet insufficiently organised units. These results indicate a need for monitoring the implementation of the basic standards in stroke units. PMID- 12649608 TI - Assessment of regional acute stroke unit care in Italy: the PROSIT study. AB - BACKGROUND/OBJECTIVES: The aim of this study was to survey acute in-hospital stroke services in seven Italian regions. METHODS: Regional hospital discharge registers were searched to identify hospital services that record at least 50 acute stroke discharges per year and the doctors in charge of these services were interviewed. RESULTS: Only 31 (7%) out of 447 services could be classified as stroke units (SUs). Of these, only 9 (29%) were found to have vital function monitoring systems available for all patients. The most striking differences between SUs and mixed stroke services emerged when comparing staff/patient ratios in the two types of unit. CONCLUSIONS: There are still not enough SUs in Italy, and fewer than 10% of stroke patients can be admitted to such units. PMID- 12649609 TI - Stroke units - the Norwegian experience. AB - The main aims of acute stroke care should be to reduce death and disability. Treatment in stroke units is the only treatment option proven to reduce death in acute stroke patients. Stroke unit care is also the only treatment which has shown a marked reduction in disability for general stroke patients, and also the only treatment with beneficial long-term effects on survival, disability and quality of life. Most acute hospitals in Norway have established stroke units. The recommended stroke unit model is a non-intensive combined unit able to focus simultaneously both on acute care and rehabilitation. This model has been very well evaluated in randomised trials and has achieved some of the best results regarding acute stroke care. Hence one can probably conclude that stroke unit care in Norway is evidence based. PMID- 12649610 TI - Stroke units and stroke services in Portugal. AB - Stroke is the first cause of death in Portugal. In 1999 more than 70,000 patients with a stroke were admitted to public hospitals, with total hospitalisation costs of EUR 188 million. Stroke patients are traditionally treated in the departments of medicine and/or neurology. In 2001 the Department of Health approved a rapid transport system of acute stroke victims to the stroke units as well as 'Stroke Unit Guidelines'. The aim of these guidelines is to define norms facilitating the launch of stroke units in all Portuguese hospitals admitting more than 300 stroke patients a year. By July 2002, five stroke units were already operating. PMID- 12649611 TI - Stroke units in Hungary - the Debrecen experience. AB - The Debrecen Stroke Unit covers a catchment area of 210,000 inhabitants in eastern Hungary. The unit was established at the Department of Neurology of the University Hospital in 1974 and has 23 beds, 7 of which have monitoring facilities. The unit treats about 600 patients with acute cerebrovascular diseases annually - about 60% of all hospitalised stroke cases in the region. Overall, 18 registered nurses and 4 nurse helpers work for the unit. Computer tomography is performed in over 90% of cases. Carotid duplex ultrasound and echocardiography are part of the routine examinations in ischaemic strokes. Delay from onset of stroke to hospital arrival is the main barrier against the use of rt-PA. Average length of stay is 12 days; a lack of rehabilitation and nursing capacities sometimes delays discharge of dependent patients. The hospital is reimbursed the costs of stroke care based on DRG. PMID- 12649612 TI - Stroke services in Scotland. AB - Stroke services are developing rapidly in Scotland. Most of the 33 hospitals which admit stroke patients from the community have a stroke unit and access to brain imaging. However, acute stroke units are few and many of the 10,000 patients each year still do not access a stroke unit bed. A recently introduced National Strategy aims to ensure that all patients have access to early stroke unit care, brain imaging and appropriate specialist rehabilitation and secondary prevention. A national system to monitor the progress of the strategy is being developed. PMID- 12649613 TI - Acute stroke units in Austria are being set up on a national level following evidence-based recommendations and structural quality criteria. AB - A system of acute stroke units is being set up in Austria, which will care for 70% of all acute strokes by 2005. This nationwide project has been planned according to evidence-based principles and contains pre-specified structural components for acute stroke care. With some exceptions in remote mountain regions, all stroke units can be reached within 90 min from any community. All units are within neurological departments. An ongoing documentation of quality performance shows that these units are being well accepted by the general population and the medical community alike. PMID- 12649614 TI - Metabolic acidosis of chronically hemodialyzed patients. AB - Metabolic acidosis is a condition that is commonly encountered in both chronic renal failure and in end-stage renal disease. Metabolic acidosis is associated with many adverse effects: negative nitrogen balance, increased protein decomposition, anorexia, fatigue, bone lesions, impaired function of the cardiovascular system, impaired function of the gastrointestinal system, hormonal disturbances, insulin resistance, hyperkalemia, altered gluconeogenesis and triglyceride metabolism, increased progression of chronic renal failure, and growth retardation in children. Even 'minor' degrees of metabolic acidosis are deleterious. Metabolic acidosis of end-stage renal patients could be successfully corrected with bicarbonate hemodialysis and with peroral bicarbonate-containing phosphate binders, i.e. calcium carbonate. Bicarbonate powder compared with bicarbonate solutions has some advantages and enables a stabile composition of electrolytes. 'High' dialysate bicarbonate (40- 42 mmol/l) is a safe, well tolerated and useful tool for better correction of the metabolic acidosis and must become a standard of hemodialysis treatment. Measured postdialysis blood bicarbonate concentration should be obtained at least every month and correction of metabolic acidosis by maintaining serum bicarbonate >or=22 mmol/l should be a goal of the management of patients undergoing chronic hemodialysis. PMID- 12649615 TI - An algorithm for train-of-four monitoring in patients receiving continuous neuromuscular blocking agents. AB - Neuromuscular blocking agents (NMBA) are increasingly being used in the management of critically ill patients. These medications are used to facilitate mechanical ventilation. Strange and colleagues estimated that only 4% of intensive care units utilize train-of-four (TOF) monitoring on a regular basis and that 70% never use this type of monitoring. The ease of use and low cost of the equipment lends itself to the increasingly frequent recommendations for this type of monitoring. Although unwilling to go so far as to establish this as a standard of care, the American College of Critical Care Medicine of the Society of Critical Care Medicine released an executive summary recommending train-of four monitoring for patients receiving continuous or sustained NMBAs. This protocol and algorithm was developed to facilitate the objective monitoring of patients receiving continuous NMBAs. PMID- 12649616 TI - The use of propofol in the mechanically ventilated medical/surgical intensive care patient: is it the right choice? AB - This article provides a review of the use of propofol in the mechanically ventilated patient. The indications for the administration of propofol are presented as well as the contraindications, adverse effects, and nursing care. PMID- 12649617 TI - Bypass injury: implications of cardiopulmonary bypass. AB - Despite significant advances made in cardiac surgery over the last 2 decades, the process of bypassing the heart and lungs or cardiopulmonary bypass (CPB) continues to precipitate physiologic insults that have an impact on a patient's ability to recover from open-heart surgery. This article explores some of these potential insults and reviews some of the interventions used to neutralize the impact of CPB. PMID- 12649618 TI - Ethical analysis. AB - Critical care nurses care for patients at the end of their lives on a daily basis. Ethical dilemmas are often encountered while caring for these patients and their families. This article reviews a case scenario along with a framework for ethical analysis. PMID- 12649619 TI - Assessing acute and critically ill patients for problem drinking. AB - More critically ill patients have acute drinking problems than chronic problems with alcohol dependence. A hospitalization offers an opportunity for nurses to perform a brief assessment and initiate a simple intervention to advise patients to reduce their drinking. A few minutes of advice may make a long-term difference in a patient's health. PMID- 12649620 TI - Sexual concerns of patients and partners after an implantable cardioverter defibrillator. AB - Sexual concerns of patients after an implantable cardioverter defibrillator (ICD) exist, but are not well understood. This study explored sexual concerns and educational needs of those living with an ICD and their partners. Study results suggest a need for written patient education tools specific to sexual issues for patients and partners, and educational resources for health professionals. Addressing the fears and concerns related to sexual function of ICD patients and partners is an essential aspect of rehabilitation and recovery. PMID- 12649622 TI - A closing word: tidbits about Florence Nightingale. PMID- 12649623 TI - Compliance with FDA recommendations for pemoline. PMID- 12649624 TI - Venlafaxine and increased aggression in a female with autism. PMID- 12649625 TI - Prazosin in PTSD. PMID- 12649626 TI - Youth suicide risk and preventive interventions: a review of the past 10 years. AB - OBJECTIVE: To review critically the past 10 years of research on youth suicide. METHOD: Research literature on youth suicide was reviewed following a systematic search of PsycINFO and Medline. The search for school-based suicide prevention programs was expanded using two education databases: ERIC and Education Full Text. Finally, manual reviews of articles' reference lists identified additional studies. The review focuses on epidemiology, risk factors, prevention strategies, and treatment protocols. RESULTS: There has been a dramatic decrease in the youth suicide rate during the past decade. Although a number of factors have been posited for the decline, one of the more plausible ones appears to be the increase in antidepressants being prescribed for adolescents during this period. Youth psychiatric disorder, a family history of suicide and psychopathology, stressful life events, and access to firearms are key risk factors for youth suicide. Exciting new findings have emerged on the biology of suicide in adults, but, while encouraging, these are yet to be replicated in youths. Promising prevention strategies, including school-based skills training for students, screening for at-risk youths, education of primary care physicians, media education, and lethal-means restriction, need continuing evaluation studies. Dialectical behavior therapy, cognitive-behavioral therapy, and treatment with antidepressants have been identified as promising treatments but have not yet been tested in a randomized clinical trial of youth suicide. CONCLUSIONS: While tremendous strides have been made in our understanding of who is at risk for suicide, it is incumbent upon future research efforts to focus on the development and evaluation of empirically based suicide prevention and treatment protocols. PMID- 12649627 TI - Developing strategies for psychopharmacological studies in preschool children. AB - OBJECTIVE: To identify the obstacles and special challenges-ethical, practical, scientific, and regulatory-faced by investigators who attempt to conduct psychopharmacological studies in preschoolers. METHOD: In a workshop held at the 47th Annual Meeting of the American Academy of Child and Adolescent Psychiatry, featuring interactive sessions designed to elicit discussion of the theory and feasibility of research in this young population, several key domains were identified: diagnosis and assessment, ethics, research design, special considerations for preschoolers, regulatory/industry issues, and education/training. RESULTS: A Pediatric Psychopharmacology Initiative is needed to consolidate recommendations from this and other workshops and current federal, research, and regulatory committees. A scholarly review and a guide for institutional review boards and investigators should be prepared on issues related to preschoolers. Developmental specialists provide valuable expertise that can strengthen studies of pediatric psychopharmacology. "N of 1" case studies can provide valuable information to clinicians. Only preschoolers with severe symptoms that occur in several interpersonal contexts should be entered into trials. Indications for the study of symptom complexes (e.g., aggression) rather than specific diagnoses should be examined and considered for regulatory activities. Psychopharmacology practice parameters for preschoolers are needed. CONCLUSIONS: With preschoolers being increasingly treated with psychopharmacological agents, the need for investigations to address the safety and efficacy of these medications is becoming a central issue for researchers from many disciplines. PMID- 12649629 TI - ADHD treatment with once-daily OROS methylphenidate: interim 12-month results from a long-term open-label study. AB - OBJECTIVE: Few treatment studies of attention-deficit/hyperactivity disorder (ADHD) extend beyond a few months. This article reports an interim analysis of a 24-month study evaluating the 12-month tolerability and effectiveness of a once daily OROS formulation of methylphenidate (OROS MPH) in children with ADHD. METHOD: Children, aged 6-13 years, with ADHD who participated in previous controlled studies and were MPH responders, received once-daily OROS MPH in this multicenter, open-label, nonrandomized study. Effectiveness was evaluated monthly by parents/caregivers and schoolteachers using validated rating scales (e.g., IOWA Conners). Safety and adverse events assessments involved objective (e.g., vital signs, growth) and subjective (sleep quality, tics) reporting. RESULTS: Seventy-one percent of subjects (289/407) completed 12 months' treatment. Effectiveness was maintained throughout 12 months as demonstrated by stable IOWA Conners ratings and sustained improvements in peer interaction and Global Assessment Scale scores. OROS MPH was well tolerated, with adverse events similar to those expected with short-acting stimulant medication. OROS MPH had minimal impact on sleep quality and tics. There were no clinically meaningful changes in blood pressure, pulse, or height. The apparent absence of meaningful changes is tempered by the fact that children were MPH responders and were medicated at baseline, most for extended periods prior to enrollment. CONCLUSION: In this open label study, once-daily OROS MPH treatment appears to be well tolerated and effectiveness was maintained for up to 12 months in these children with ADHD. PMID- 12649628 TI - Fluoxetine for the treatment of childhood anxiety disorders. AB - OBJECTIVE: To assess the efficacy and tolerability of fluoxetine for the acute treatment of children and adolescents with generalized anxiety disorder, separation anxiety disorder, and/or social phobia. METHOD: Anxious youths (7-17 years old) who had significant functional impairment were randomized to fluoxetine (20 mg/day) (n = 37) or placebo (n = 37) for 12 weeks. RESULTS: Fluoxetine was effective in reducing the anxiety symptoms and improving functioning in all measures. Using intent-to-treat analysis, 61% of patients taking fluoxetine and 35% taking placebo showed much to very much improvement. Despite this improvement, a substantial group of patients remained symptomatic. Fluoxetine was well tolerated except for mild and transient headaches and gastrointestinal side effects. Youths with social phobia and generalized anxiety disorder responded better to fluoxetine than placebo, but only social phobia moderated the clinical and functional response. Severity of the anxiety at intake and positive family history for anxiety predicted poorer functioning at the end of the study. CONCLUSIONS: Fluoxetine is useful and well tolerated for the acute treatment of anxious youths. Investigations regarding the optimization of treatment to obtain full anxiety remission and the length of treatment necessary to prevent recurrences are warranted. PMID- 12649630 TI - Perceptions of P.R.N. psychotropic medications by hospitalized child and adolescent recipients. AB - OBJECTIVE: To understand patient perceptions of p.r.n. medication for agitation used in institutional settings. METHOD: A questionnaire was administered to child and adolescent psychiatric inpatients within 12 hours of their receiving p.r.n. medication for agitation and repeated 4 hours later. Research staff completed a form to rate each patient's comprehension and behavior in completing the questionnaire. RESULTS: Forty-two patients completed the survey: 38 males and 4 females, aged 7 to 17 years (mean age 12.19 years, median age 12 years). Patients requested the p.r.n. medication in 30% of episodes and assisted with the decision in an additional 10%. The p.r.n. was perceived for loss of control in more than half the cases. About 50% of the youngsters felt that the p.r.n. they received was the "best" for them. Only 30% answered that "something" could have been done by themselves or by staff to avoid requiring a p.r.n. medication. About 65% felt that "something good" happened from receiving the medication. The test-retest reliability of all but one question by kappa statistic was p 3.5); therefore, no study had to be repeated. CONCLUSION: Routine skeleton images with 50% dose reduction yield good image quality and good clinical acceptance. In cases with abundant soft tissue, less dose reduction or standard dose is required. PMID- 12649648 TI - Menstrual-cycle dependence of breast parenchyma elasticity: estimation with magnetic resonance elastography of breast tissue during the menstrual cycle. AB - RATIONALE AND OBJECTIVES: Magnetic resonance elastography (MRE) is a promising diagnostic method that produces images with a contrast proportional to the elasticity of the tissue. This study investigated using MRE the dependence of breast tissue elasticity from the menstrual cycle of healthy volunteers. METHODS: Five volunteers (age 26-36) without breast disease and contraceptive medication were examined once weekly over 2 menstrual cycles. Examinations were performed with a 1.5 T magnet (ACS-NT, Philips Medical Systems, Best, The Netherlands). Low frequency mechanical waves (65 Hz) were transmitted into the tissue by an oscillator. By means of a motion-sensitive spin-echo sequence, mechanical waves were displayed within the phase of the MR image and phase images were used to reconstruct the local distribution of elasticity. The elasticity of fibroglandular tissue and adipose breast tissue was analyzed individually, and the median and mean values of elasticity over the menstrual cycle were determined. RESULTS: All volunteers presented a repeating pattern concerning the elasticity over the 2 cycles. After 5 days of the onset of menses, the median value of elasticity for fibroglandular adipose tissue declined significantly by 29% (P = 0.010). After the second week of the cycle, fibroglandular tissue showed again an increase in elasticity (P = 0.028). The highest median values of elasticity were obtained during days 11 to 23 with an increase of up to 35%. For adipose tissue, only a slight and not significant variation of elasticity during the menstrual cycle was determined. CONCLUSION: MRE is able to measure a dependence of tissue elasticity on the menstrual cycle. PMID- 12649649 TI - Outpatients with low back pain: an analysis of the rate per day of pain improvement that may be expected and factors affecting improvement. AB - OBJECTIVE: To determine, only for those who improved, the rate per day of improvement expected for outpatients with low back pain and to identify factors influencing pain improvement: pain duration before assessment, pain and physical functioning levels before assessment, age, sex, and affective factors. It was presupposed that affective factors would have an appreciable effect. DESIGN: The LIFEware System database was used. Analysis was performed on 1292 records for the Painfree measure and 1562 records for the LIFEware System Visual Analog Scale. Measures for pain, physical functioning, and affective well-being were analyzed for 0-30 days vs. >30 days of pain duration before assessment using classification and regression trees analysis. RESULTS: In both Painfree and LIFEware System Visual Analog Scale, 73% improved and 27% did not improve. Of those who improved, outpatients with 0-30 days of pain duration before assessment had higher per day rates of improvement than the >30 days group. Factors affecting improvement were, in descending order, more initial pain, younger age, and positive affective well-being; physical functioning did not affect rate of improvement. Factors affecting improvement for outpatients with >30 days since onset were, in descending order, more initial pain and better initial physical functioning; age and affective well-being were not factors. For all, neither sex nor the "satisfaction with life in general" question affected low back pain rate of improvement. CONCLUSION: Findings may be useful for clinical application because the actual rate of improvement may be compared with the expected rate. There was only a weak relationship shown between affective factors and pain improvement. PMID- 12649650 TI - Percentage of body weight carried by students in their school backpacks. AB - OBJECTIVE: To determine the percentage of body weight represented by backpacks and the types of backpacks carried by elementary school students. DESIGN: A cross sectional study was conducted in three elementary schools in central Texas. All students in kindergarten through fifth grade were invited to participate. A letter, with no direct mention of backpack weighing, was sent to each parent for written consent. Student weights and heights and their backpack weights were measured by trained research assistants. RESULTS: The mean backpack weights, which increased significantly with increasing grade level, varied significantly by school, backpack type, day of week, body mass index, and race/ethnicity. The backpack loads represented an average of 8.2% (95% confidence interval, 7.8-8.5) of student body weights but increased significantly with increasing grade level, from 6.2% among kindergarteners to 12.0% among fifth graders. Twenty-six percent of students carried backpacks that weighed at least 10% of their body weights. A total of 25 students (3.5%) had backpacks with wheels. Wheeled backpack users were significantly older and more likely to be girls or from school B. Concern about weight was the most popular reason cited for using a wheeled backpack. CONCLUSIONS: Backpack loads represent a significant percentage of body weight of this sample of students aged 5-12 yr. Some students are already using wheeled backpacks, which may have their own hazards. PMID- 12649651 TI - Reliability of the Catz-Itzkovich Spinal Cord Independence Measure assessment by interview and comparison with observation. AB - OBJECTIVE: To examine the reliability of assessment with the Catz-Itzkovich Spinal Cord Independence Measure II (SCIM II) by interview and compare the findings with assessment by observation. DESIGN: In a cohort, comparative study, 28 inpatients with spinal cord lesions were assessed by two nurses using the Catz Itzkovich SCIM II (interview) and by a multidisciplinary team (observation). RESULTS: Total agreement between interviewers ranged from 50% to 80% (Kappa coefficients 0.40-0.60). Pearson's coefficients of the correlation between scores obtained for the various SCIM subscales by interview or observation were 0.765 0.940 (P < 0.0001). The differences in mean scores obtained between the interview and observation methods were small and not statistically significant for most of the subscales. CONCLUSIONS: The results support the reliability of the Catz Itzkovich SCIM assessment by interview and show it to be comparable with assessment by observation. The SCIM II interview may serve as an accurate measure of daily function in patients with spinal cord injury. However, with the sample of the study being relatively small, a larger scale examination is needed to generalize the results. PMID- 12649652 TI - Clinical prediction of falls in the elderly. AB - OBJECTIVE: To assess the ability of physical and occupational therapists engaged in rehabilitation of the elderly to predict posttreatment falls. DESIGN: Prospective cohort study of 15 mo in duration at an urban academic medical center rehabilitation unit. A total of 165 consecutively admitted geriatric individuals were rated for fall risk by 14 physical and seven occupational therapists. Measurements included the Mini-Mental State Examination, Geriatric Depression Scale, FIM, and therapists' ratings of fall likelihood. RESULTS: Both disciplines evidenced an ability to predict who would fall in the 3 mo after discharge. Clinical judgment regarding fall risk, however, added little value over two major predictors of future falls, fall history and the presence of a neurologic condition. CONCLUSION: Trying to predict an infrequent future event such as falls is inherently difficult. Education regarding known fall-risk factors and inclusion of standardized measurements of physical status are recommended to potentially improve rates of detection, along with adoption of a realistic attitude regarding our abilities to forecast infrequent events. PMID- 12649653 TI - Does balance or motor impairment of limbs discriminate the ambulatory status of stroke survivors? AB - OBJECTIVE: This study was performed to determine if ambulatory function is governed by motor impairment of limbs or balance ability in subjects with hemiplegia caused by stroke. DESIGN: Seven patients who walked with physical assistance (FIM 4) after stroke and 13 who walked independently with assistive devices (FIM 6) were compared with 13 healthy subjects. Motor impairment of limbs was evaluated with the Fugl-Meyer Assessment. The Berg Balance Scale and limit of stability test of the Smart Balance Master were used to evaluate balance ability. RESULTS: The FIM 6 group and the controls were best differentiated by motor impairment of the paretic limbs and limit of stability in the backward direction. Motor impairment of the upper limb and limit of stability in direction toward the paretic side separated the FIM 4 from the FIM 6 group. Upper limb motor impairment and the Berg Balance Scale consistently separated the three subject groups. CONCLUSIONS: Motor impairment in the paretic upper limb and balance dysfunction should be addressed in treatments working toward independent ambulation. PMID- 12649654 TI - Effect of botulinum toxin type A on cerebral palsy with upper limb spasticity. AB - OBJECTIVE: The objective of this study was to investigate the effects of botulinum toxin type A injections in reducing upper limb muscular spasticity and in improving motor function in children with cerebral palsy. DESIGN: Fifteen children with spastic cerebral palsy who were undergoing regular physical and occupational therapy were enrolled. Botulinum toxin type A injections in clinically indicated target muscle groups were administered after the children had received 3 mo of therapy. A follow-up study was carried out at 6 wk and 12 wk, respectively, after the botulinum toxin type A injections. The main outcome measurements included the Modified Ashworth Scale, the upper limb Physician's Rating Scale, the Bruininks-Oseretsky Test of Motor Proficiency, and the self care domain of the Pediatric Evaluation of Disability Inventory. RESULTS: The reduction of spasticity in the treated muscle groups differed significantly between the control period and both study periods. Improvements on the Physician's Rating Scale score during the study period also differed significantly as compared with improvements during the control period. There was a significant difference in the improvement of fine motor skills, as measured with Bruininks-Oseretsky Test of Motor Proficiency, between the control period and both study periods. Improvements in self-care capability differed significantly between the control period and 12 wk after botulinum toxin type A treatment, but not between the control period and at 6 wk after treatment. Muscle strength of grasp and pinch did not differ significantly between the control and the study period. Distribution of body parts involvement, disease severity, and function in daily living activities had no significant correlation with functional improvement after the treatment. CONCLUSIONS: Our findings support the premise that botulinum toxin type A injections are effective in reducing upper limb spasticity and in improving movement pattern and fine motor function of patients with spastic cerebral palsy. A reduction in caregivers' burden and improved quality of life were demonstrated through the study period. PMID- 12649655 TI - Effect of counterforce forearm bracing on wrist extensor muscles performance. AB - OBJECTIVES: We examined (1) the effect of a counterforce forearm brace on isokinetic strength, stretch reflex, passive stretching, and proprioception of the forearm muscles in healthy subjects and (2) the effect of different strap tensions of the brace on the above variables. DESIGN: The dominant hand of 15 healthy subjects were tested under four randomized conditions: (1) no brace, (2) brace with minimal tension, (3) brace with 2.5 kg force tension, and (4) brace with 5 kg force tension. The tests included isokinetic wrist extensors strength, passive stretching to the wrist extensors until onset of pain, joint proprioception, and reflex latency of forearm extensor muscles. A repeated measures multivariate analysis of variance test was used to analyze the data, and significant results were further analyzed with post hoc linear contrasts with alpha at 0.0083. RESULTS: There was no difference in isokinetic strength, proprioception, and stretch reflex latency among the four conditions. For passive stretching, there was a significant difference (P = 0.001) in that using a brace produced a higher pain threshold than without a brace. CONCLUSION: A forearm counterforce brace has no effect on strength, proprioception, and stretch reflex latency, but it increases the pain threshold. PMID- 12649656 TI - Reference values for peroneal nerve motor conduction to the tibialis anterior and for peroneal vs. tibial latencies. AB - OBJECTIVE: To generate a large normative database for the peroneal study to the tibialis anterior and to analyze differences between the peroneal latency to the extensor digitorum brevis and the tibial latency to the abductor hallucis. DESIGN: A total of 244 asymptomatic subjects without risk factors for neuropathy were tested for latency, amplitude, area, duration, and nerve conduction velocity to the tibialis anterior. For the peroneal/tibial analysis, 221 subjects were included. Temperature control was used, and demographic characteristics were recorded. RESULTS: Latency to the tibialis anterior increased and nerve conduction velocity decreased with increasing height. Amplitudes, area, and nerve conduction velocity were lower in older persons. The upper limits of normal (97th percentile) were 4.9 msec (latency) and 15.0 msec (duration). The lower limits of normal (3rd percentile) were 1.7 mV (amplitude), 6.8 microV/sec (area), and 43 m/sec (nerve conduction velocity). The upper limits of normal side-to-side difference was 1.2 msec (latency), 6.3 msec (duration), 50% (amplitude), 47% (area), and 20% (nerve conduction velocity). The upper limits of normal drop from below- to above-fibular-head stimulation was 36% (amplitude) and 43% (area). For the peroneal/tibial comparison, the upper limits of normal latency increase of peroneal over tibial was 1.8 msec and tibial over peroneal was 1.5 msec. CONCLUSION: A large normative database was collected. Upper and lower limits of normal were calculated, and side-to-side differences were analyzed. For the peroneal/tibial comparison, an upper limits of normal difference was described. PMID- 12649657 TI - National variability in permanent partial impairment ratings. AB - OBJECTIVE: To evaluate the variability in permanent partial impairment ratings of three different clinical vignettes by physiatrists throughout the United States with regard to location in the country, years of experience, and type of practice. DESIGN: Participants were asked to fill out a questionnaire requesting their state of practice, number of years in practice, scope of their practice, and percentage of their time devoted to worker's compensation cases. They were also given three clinical scenarios and were asked for the diagnosis of each, along with an impairment rating and how that rating was obtained. RESULTS: The range of impairment ratings for the case of radiculopathy was 0-14%. The range for the case of radial neuropathy was 3-60%. The range for the total hip replacement case was 8-100%. The responses did not correlate with region, years of experience, or type of practice except in the first case, in which a trend was noted that the younger respondents gave lower ratings. Participants used different sources for obtaining their results, which included the third, fourth, or fifth edition of the American Medical Association's guides to impairment. Some used their state guides, whereas others used their clinical judgment. CONCLUSION: There is a great deal of variability of permanent partial impairment ratings throughout the country; several different sources are used to determine ratings. PMID- 12649658 TI - Sagittal plane loading response during gait in different age groups and in people with knee osteoarthritis. AB - OBJECTIVE: To investigate the gait patterns and the sagittal ground reaction forces in different age groups and in people with knee osteoarthritis. DESIGN: Motion analysis and force platform data were collected for a total of 55 female subjects capable of independent ambulation. Subjects were divided into three groups consisting of the control group, the elderly group, and the osteoarthritis knee group. Gait parameters of walking velocity, cadence, step length, stride time, single- and double-support time, and sagittal ground reaction forces were obtained during comfortable walking speed. Gait analysis was performed in a tertiary hospital's gait laboratory. Variables were analyzed using a univariate repeated-measures analysis of variance. Statistical significance was set at a value of P < 0.05. RESULTS: The osteoarthritis knee group had slower walking velocity, lower cadence, and longer stride time as compared with the elderly and young control groups (P < 0.05). In ground reaction force studies, the first peak time, expressed in percentage of gait cycle, was significantly longer in the osteoarthritis knee group (20.8 +/- 3.2) as compared with the elderly (17.8 +/- 2.0) and young control groups (17.1 +/- 1.8, P < 0.01). The force during time of minimal midstance was larger in the osteoarthritis knee group (90.9 +/- 5.3) as compared with the elderly and young control groups (P < 0.05). The second peak force was significantly smaller in the osteoarthritis knee group as compared with the young control group (P < 0.01). The force change in the midfoot region in the osteoarthritis knee and elderly groups revealed more loading force onto the midfoot region during midstance as compared with the young control group (P < 0.01). CONCLUSION: Gait parameters in the elderly and osteoarthritis knee patients were characterized by slower walking velocity, lower cadence, shorter step length, longer stride time, and longer double-support time. Less heel contact and push-off forces were noticed in these two groups, with more loading force onto the midfoot during midstance. PMID- 12649659 TI - Spinal cord injury computer-assisted instruction for medical students. AB - OBJECTIVE: To determine if a computer-assisted instruction program would improve fourth-year medical students' knowledge base related to spinal cord injury, as determined by pretest and posttest scores. DESIGN: A multimedia computer-assisted instruction program was developed and offered on a volunteer basis to an entire class of fourth-year medical students (n = 168). Effectiveness of the instructional content was evaluated with pretests and posttests, and overall user satisfaction with the module was assessed with the courseware evaluation. RESULTS: The responses yielded 83 sets of completed pretest and posttest pairs and 80 sets of fully completed courseware evaluations. Mean posttest score was significantly higher than the mean pretest score (pretest, 6.65 +/- 1.44, vs. posttest, 7.36 +/- 1.38; df = 82, t = -4.74, P < 0.001). Courseware evaluations yielded positive ratings in all areas, including applicability and usability. CONCLUSIONS: The significantly increased posttest scores suggest that the students left the program with an expanded knowledge base in the content areas of spinal cord injury medicine covered in the computer-assisted instruction program. PMID- 12649660 TI - Effects of regular exercise training on skeletal muscle contractile function. AB - Skeletal muscle function is critical to movement and one's ability to perform daily tasks, such as eating and walking. One objective of this article is to review the contractile properties of fast and slow skeletal muscle and single fibers, with particular emphasis on the cellular events that control or rate limit the important mechanical properties. Another important goal of this article is to present the current understanding of how the contractile properties of limb skeletal muscle adapt to programs of regular exercise. PMID- 12649661 TI - Their subjective condition will be worsened? PMID- 12649662 TI - Much ado about getting a "DO"! Beauty parlor stroke syndrome and sink radiculopathy: a redux. PMID- 12649664 TI - Challenges in the early diagnosis and staging of Fallopian-tube carcinomas associated with BRCA mutations. AB - The histopathologic diagnosis of fallopian-tube carcinoma has been traditionally made at an advanced stage. More recently, predictive genetic BRCA testing is leading to the recognition in prophylactic oophorectomy specimens of clinically occult tubal carcinomas that are frequently in situ or small early-stage invasive carcinomas. These early lesions present a challenge in diagnosis and staging because the available criteria for the histopathologic diagnosis and staging of tubal carcinoma were derived from the clinicopathologic experience derived from the usual high-stage tubal carcinomas. The detection of early-stage tubal carcinomas requires that all tubal tissue be submitted for histologic examination. The diagnostic criteria for tubal in situ carcinoma have been defined, although the natural history of this lesion is unclear. Similarly defined criteria for a diagnosis of tubal dysplasia are lacking. Any early, invasive tubal carcinoma should be staged using a refined staging system suitable for early stage and fimbrial carcinomas. The adoption of these methods should increase our knowledge of early-stage tubal carcinoma and may add to our understanding of the development of ovarian-epithelial neoplasia. PMID- 12649665 TI - Mesonephric remnants or hyperplasia can cause abnormal pap smears: a study of three cases. AB - Three postmenopausal women were found to have abnormal Pap smears attributable to cells derived from mesonephric remnants or mesonephric hyperplasia of the cervix (one patient) or the upper vagina (two posthysterectomy patients). Mesonephric remnants or hyperplasia may represent the sole cause of abnormal Pap smears. PMID- 12649666 TI - Pagetoid squamous cell carcinoma in situ (pagetoid Bowen's disease) of the external genitalia. AB - Approximately 5% of cutaneous squamous cell carcinomas in situ (SCCIS) have a nested pattern, referred to as pagetoid SCCIS, or pagetoid Bowen's disease. This growth pattern may simulate extramammary Paget's disease (EPD) when the external genitalia are involved. We report two cases of genital pagetoid SCCIS, including the first example affecting the vulva. Using several known cases each of classic (bowenoid) SCCIS of the vulva, primary cutaneous vulvar EPD, and cutaneous melanoma in situ as controls, we performed a battery of immunohistochemical and mucin stains to study the phenotype of pagetoid SCCIS. Both cases of pagetoid SCCIS were strongly positive for cytokeratin (CK) 7, a highly sensitive and popular marker for EPD. Stains with the high molecular weight cytokeratin marker Keratin-903 (34betaE12) showed 4+ immunoreactivity, although not with the intensity seen in the juxtaposed normal keratinocytes or in the cells of classic SCCIS. Immunoreactivity scores were 2+ for CK5/6, 2+ for CK19, and 1+ to 3+ for CK 13. Unlike the Paget's cells of EPD, the nested cells of pagetoid SCCIS were devoid of mucin and were nonimmunoreactive with GCDFP-15, CEA, CAM5.2, and c erbB2. Stains for CK20, S-100, and Melan A also were negative. Although CK7 is a sensitive marker for the Paget's cells of EPD, this study corroborates the unexpected CK7 positivity in pagetoid SCCIS, precluding its usefulness in distinguishing these two diseases. Combined with mucin stains, a limited immunohistochemical panel may be of diagnostic value in particularly difficult cases. PMID- 12649667 TI - Metastatic melanoma to the vagina: clinicopathologic and immunohistochemical study of three cases and literature review. AB - We report the clinicopathologic features of three cases of malignant melanoma metastatic to the vagina; only one similar case has been previously reported. The age of the patients ranged from 33 to 70 years; two presented with vaginal bleeding. Two patients had a history of a previous primary malignant melanoma, one a preauricular melanoma treated 7 years earlier and the other a vulvar melanoma treated 2 years earlier. In the third case, a primary malignant melanoma was found on the sole of the right foot after the patient had presented with the vaginal metastases. On gross examination, the vaginal tumors were polypoid; two were on the posterior wall and one was on the anterior wall. All were of epithelioid cell type and were positive with S-100 and microphthalmia transcription factor. Tyrosinase was positive in two cases, HMB-45 was positive in one case, and MART-1 was focally positive in one case. One patient underwent wide local excision of tumor followed by chemotherapy, one patient had intracavitary radiotherapy, and one had palliative radiotherapy to the brain only. The patients died after a follow-up of 3, 14, and 48 months. PMID- 12649668 TI - Loss of expression of MLH1, MSH2, MSH6, and PTEN related to endometrial cancer in 68 patients with endometrial hyperplasia. AB - Derangements in the tumor suppressor gene PTEN and the mismatch-repair genes, hMLH1, hMSH2, and hMSH6, have an important role in endometrial carcinogenesis. The purpose of this study was to assess immunohistochemically the pattern of protein expression for these genes in 68 patients with endometrial hyperplasia and to determine the relation of protein expression to cancer development or coexistence of cancers. Loss of expression of these genes also was evaluated as potential tumor markers for clinical use. PTEN and hMLH1 both showed loss of expression in 55% of specimens from 18 patients with subsequent or coexisting carcinoma. D&C specimens from 50 patients who did not develop cancer (10 patients underwent hysterectomy within 2 years; 40 had no hysterectomy; follow-up of 10-20 years), expressed protein at a much higher frequency (92% for PTEN and 98% for hMLH1). The parameter with the strongest independent relation to subsequent or coexisting carcinoma in a stepwise multiple logistic regression analysis was hMLH1. Evaluation of the investigated factors as prognostic markers for tumor development showed high specificity (92% for PTEN, 98% for MLH1) at the expense of sensitivity (56% for PTEN, 56% for MLH1). The results were compared with the results of the computerized image analysis algorithm, the D-score. PMID- 12649669 TI - C-kit immunoreactivity in endometrial adenocarcinomas and its clinicopathologic significance. AB - The proto-oncogene c-kit is a transmembrane-tyrosine-kinase receptor that is structurally related to the platelet-derived growth-factor receptor (PDGFR) and is involved in cell differentiation. C-kit has been found to be expressed in certain solid tumors and may play a role in their tumorigenesis. Recently, a tyrosine-kinase inhibitor specific for the PDGFR family, bcr-abl, and c-kit (STI571) has been reported to have therapeutic effects in tumors expressing the aberrant forms or high quantities of target proteins. Expression of c-kit has not been well evaluated in endometrial adenocarcinomas. In this study, c-kit immunoreactivity was evaluated on paraffin sections of 72 endometrial adenocarcinomas (57 endometrioid, 10 serous, and 5 clear cell) with a polyclonal antibody. Immunoreactivity of c-kit was analyzed semiquantitatively and correlated with various clinicopathologic factors. Cytoplasmic c-kit immunoreactivity was detected in 42 (58%) tumors. Thirty-four (60%) endometrioid, 8 (80%) serous, and 0 of the 5 clear-cell adenocarcinomas were c-kit positive. There was a significant correlation between c-kit positivity and the depth of myometrial invasion. Patients with c-kit-positive endometrial adenocarcinomas more frequently had metastases and shorter disease-free survival. Expression of c kit may be a potentially adverse prognostic feature in endometrial adenocarcinoma. Patients with c-kit-positive advanced endometrial adenocarcinoma might benefit from tyrosine-kinase-inhibitor therapy. PMID- 12649670 TI - Expression pattern of the cell cycle promoter cyclin e in benign extravillous trophoblast and gestational trophoblastic lesions: correlation with expression of Ki-67. AB - In this study, a specific monoclonal antibody was used to immunohistochemically investigate correlated expression of the cell cycle promoter cyclin E and the proliferation marker Ki-67 in benign extravillous trophoblast and gestational trophoblastic lesions. Our data show that cyclin E is expressed in the normal extravillous trophoblast, with strongest levels of expression in the cell columns of anchoring villi. Differences could be observed in expression of Ki-67 in both normal extravillous trophoblast and gestational trophoblastic lesions. In the extravillous trophoblast of the cell columns, expression of cyclin E started more distal compared with Ki-67 and was maintained (with less intensity) into the deeper layers of interstitial trophoblast. In the benign trophoblastic lesions (exaggerated placental site [EPS] and placental site nodule [PSN]) and in the trophoblast proliferations on the surface of hydropic villi of hydatidiform moles (HM), the percentage of cells expressing cyclin E was higher than of those expressing Ki-67. The same observation could be made for a case of placental site trophoblastic tumor (PSTT). In contrast, choriocarcinomas (N=8), which are definitely malignant tumors, showed an opposite pattern, with a much higher percentage of strongly Ki-67-positive cells compared with cyclin E-positive cells. We conclude that cyclin E is expressed in benign extravillous trophoblast and gestational trophoblastic lesions, where a ratio cyclin E/Ki-67<1 characterizes choriocarcinomas, whereas PSTT and the benign lesions (HM, EPS, PSN) show expression of cyclin E in a higher percentage of cells than Ki-67 (cyclin E/Ki-67 ratio >1). PMID- 12649671 TI - Ovarian steroid cell tumors: an immunohistochemical study including a comparison of calretinin with inhibin. AB - Ovarian steroid cell tumors, not otherwise specified (SCTs, NOS) are uncommon sex cord-stromal tumors that may be difficult to distinguish from other oxyphilic or clear-cell neoplasms. Immunohistochemical staining for inhibin, although generally useful in the diagnosis of SCTs, NOS, is not positive in every case and not all laboratories have this marker available. Recently, it has been reported that calretinin is expressed by sex cord-stromal tumors. We studied six SCTs, NOS for both calretinin and inhibin expression to evaluate the sensitivity of calretinin in comparison to inhibin. We also tested for CD99, Melan-A (A103), and S-100, other markers reported to be positive in these tumors. HMB-45 and MART-1 (Ab3) completed our panel of markers. All six tumors were positive for both calretinin and inhibin. Calretinin positivity was present in 60% to >90% of tumor cells, whereas inhibin reactivity ranged from <5% to >90% of tumor cells. Membranous staining for CD99 was present in one tumor. S-100-positive cells were seen in two tumors, whereas four tumors were immunoreactive for HMB-45. All six tumors were positive for Melan-A (A103), but in general the staining was less diffuse than with calretinin. All of the tumors were essentially negative for MART-1 (Ab3). The consistent diffuse staining of the tumors in this study for calretinin, in comparison to inhibin and Melan-A (A103), suggests that it is a sensitive marker for SCTs, NOS. MART-1 (Ab3) immunostaining may be useful for cases in which melanoma is considered in the differential diagnosis. PMID- 12649672 TI - Expression and prognostic value of the cell-cycle regulatory proteins, Rb, p16MTS1, p21WAF1, p27KIP1, cyclin E, and cyclin D2, in ovarian cancer. AB - To investigate the role of cell-cycle regulatory proteins in ovarian cancer, we performed immunohistochemistry for the cell-cycle promoters cyclin E and cyclin D2 and the cell-cycle inhibitors, Rb, p16MTS1, p21WAF1, and p27 KIP1, in 93 ovarian carcinomas (77 with follow-up data). The results were correlated with clinicopathological parameters and the prognostic value was determined by multivariate analysis. Strong Rb and moderate-high cyclin E immunoreactivity in carcinomas were weakly associated with high expression of the proliferation marker Ki67. By Cox's multivariate analysis, advanced stage (p=0.013), strong Rb expression (p=0.006), and negative-weak p21 staining (p=0.011) were independent prognostic indicators of short overall survival, indicating an apparently paradoxical role of the retinoblastoma protein in these tumors. In addition, trends pointing to an association of higher age (p=0.067) and positive cyclin E immunoreactivity (p=0.093) with an unfavorable prognosis were observed. PMID- 12649673 TI - Neutral endopeptidase/CD10 expression in the stroma of epithelial ovarian carcinoma. AB - This study investigated the immunohistochemical expression and localization of neutral endopeptidase (NEP) (CD10), which plays a functional role by degrading bioactive peptides, in ovarian tumors. In normal ovaries and benign cystadenomas, NEP was not detected in any epithelial or stromal cells. In borderline tumors, NEP was detected in the stromal cells in 6 of 7 serous tumors, but not in those from mucinous tumors. In ovarian carcinomas, NEP in the stromal cells was observed in 13 of 20 serous, 8 of 10 endometrioid, and 7 of 10 clear-cell adenocarcinomas. NEP was weakly detected in only 1 of 9 mucinous adenocarcinomas. The staining intensity of stromal NEP was decreased in grades 2 and 3 serous carcinomas compared with that in grade 1 serous carcinomas. In conclusion, NEP was specifically expressed in the stroma of borderline and malignant ovarian tumors, but not in adenomas. Furthermore, stromal NEP was downregulated as the histological grade advanced. These results suggest that NEP may play a role in the regulation of neoplastic transformation and tumor differentiation in epithelial ovarian carcinomas. PMID- 12649674 TI - Lack of expression of c-kit protein (CD117) in mesenchymal tumors of the uterus and ovary. AB - c-kit is a proto-oncogene that codes for a transmembrane tyrosine kinase receptor (CD117). The gene product KIT is constitutively overexpressed in mastocytosis and gastrointestinal stromal tumors. Recently the use of the tyrosine kinase inhibitors, such as STI-571, has resulted in the successful treatment of bcr-abl positive leukemias and gastrointestinal stromal tumors. In gastrointestinal stromal tumors, immunostaining for c-kit is diffusely positive. Because the expression of c-kit in mesenchymal tumors of the uterus and ovary has not been previously studied, we evaluated its expression in 38 of these tumors by immunohistochemistry. The number of positive labeled/total tumors were as follows: 0/8 malignant mullerian mixed tumors, 4/7 ovarian fibrosarcomas, 0/1 clear-cell ovarian sarcoma, 0/4 uterine leiomyosarcomas, 1/10 low-grade endometrial stromal sarcomas, 0/2 high-grade endometrial stromal sarcomas, and 3/6 endometrial stromal nodules. In all positive cases, no more than 5% of the cells were labeled. In conclusion, unlike gastrointestinal stromal tumors, mesenchymal tumors of the uterus and ovary rarely express c-kit. Therefore, it is unlikely that patients with these tumors will benefit from treatment with the currently available tyrosine kinase inhibitors. PMID- 12649675 TI - Brenner tumor of the ovary with striking microcystic change. AB - An ovarian Brenner tumor from a 39-year-old woman exhibited striking microcystic change that was so extensive it largely obscured the nature of the tumor, causing initial diagnostic difficulty. The patient presented with symptoms related to a pelvic mass and had a unilateral, 24 cm in maximal dimension, stage I tumor that was solid and cystic. Microscopic examination revealed extensive microcystic change within the epithelial nests in large areas of the tumor such that transitional-type epithelium was only recognized very focally. The tumor had minor areas (<5%) that showed marked cytological atypia and mitotic activity, interpreted as microscopic foci of intraepithelial carcinoma, but was predominantly benign. The presence of characteristic fibromatous stroma and mucinous metaplasia within the epithelial nests provided important clues to the correct diagnosis. PMID- 12649676 TI - Urachal adenocarcinoma metastatic to the ovaries: case report and literature review. AB - We report a urachal adenocarcinoma metastatic to both ovaries in a 50-year-old Japanese woman. Pelvic examination and imaging studies revealed a large cystic tumor occupying the pelvis and another cystic tumor between the umbilicus and the urinary bladder. A laparotomy was performed. Histopathological examination revealed a urachal tumor that was a well-differentiated invasive mucinous adenocarcinoma; the overlying urothelium was intact. The right and left ovarian tumors were well-differentiated mucinous adenocarcinomas. The urachal and ovarian tumors were immunoreactive for cytokeratin 20 and carcinoembryonic antigen, but negative for cytokeratin 7. The patient is alive with lymph node and bone metastases 6 months postoperatively. This is the eighth reported case of an adenocarcinoma of the bladder with ovarian metastasis. PMID- 12649677 TI - Serous carcinoma arising in an adenofibroma of the endometrium. AB - A serous carcinoma and endometrial intraepithelial carcinoma were encountered in an endometrial adenofibroma in a 61-year-old woman. The carcinoma involved the myometrium and cervix (stage IIa). To our knowledge, this is the third documented case of an adenocarcinoma and the first serous carcinoma involving a uterine adenofibroma. PMID- 12649678 TI - Diffuse uterine adenomatoid tumor in an immunosuppressed renal transplant recipient. AB - A case of diffuse adenomatoid tumor of the uterus in an immunosuppressed renal transplant recipient is reported and compared with two previously reported, similar cases. The multinodular uterine lesion grossly resembled intramural leiomyomata except for a mucoid cut surface and a cystic serosal component. The predominant patterns of the tumor were adenoid and angiomatoid with less prominent solid and cystic patterns. Immunohistochemical and ultrastructural studies confirmed the mesothelial nature of the tumor cells. Additionally, there was strong diffuse immunoreactivity for proliferating cell nuclear antigen, a low expression of Ki-67, and weak nuclear p53 staining. The relationship between immunosuppression and diffuse adenomatoid tumors is discussed. PMID- 12649679 TI - Benign "metastasizing" lipoleiomyoma of the uterus. AB - A 62-year-old woman presented with an enlarged uterus and numerous pulmonary nodules. On histological examination, the multiple uterine and lung tumors were lipoleiomyomas and histologically identical. Tumors in both sites were immunoreactive for estrogen and progesterone receptors and had a low MIB index. Although intrapulmonary emboli from a benign lipoleiomyoma of the uterus cannot be entirely excluded, the theory of synchronous multifocal lipoleiomyomatous proliferation may provide a more satisfactory explanation in this case. PMID- 12649680 TI - Retroperitoneal primary mucinous adenocarcinoma with a mural nodule of anaplastic tumor: a case report and literature review. AB - A 38-year-old female presented with a lower abdominal mass. During the operation the mass was found to be retroperitoneal and was excised. Gross examination revealed a mucin-containing cystic lesion with a mural nodule. On microscopic examination, the cystic areas were lined by an invasive mucinous adenocarcinoma and the nodule was composed of an anaplastic sarcomatoid tumor that was immunoreactive for cytokeratin. This present case is the 21st example of a retroperitoneal primary mucinous cystadenocarcinoma and the fourth with a mural nodule. Three of four cases with a mural nodule, including our case, had a rapidly fatal outcome. PMID- 12649681 TI - Mullerianosis of the mesosalpinx: a case report. AB - We report a case of mullerianosis of the mesosalpinx, the appearance of which simulated metastatic adenocarcinoma. The patient was a 37-year-old woman with a mixed epithelial cystadenoma of borderline malignancy associated with endometriosis in both ovaries. The left mesosalpinx contained a firm, 2.2-cm, gray-white mass that on microscopic examination consisted of glands lined mostly by endocervical-type epithelium with admixed tubal-type glands and foci of endometriosis. Rupture of glands with extravasation of mucin into the surrounding stroma was observed, but there was no desmoplastic stromal reaction. Awareness of this relatively uncommon lesion is critical to avoid misdiagnosis and overly aggressive treatment. PMID- 12649682 TI - Eosinophils as a marker for invasion in cervical squamous neoplastic lesions. PMID- 12649684 TI - Voice mail, e-mail--do not delete me! PMID- 12649685 TI - Can a physician sue a state medical board under the ADA? PMID- 12649686 TI - Accountability and hospital rankings. PMID- 12649687 TI - Divided loyalties in a brain injury case. PMID- 12649689 TI - Coordination of care: the Council for Case Management Accountability's third State of the Science paper. PMID- 12649690 TI - Postacute rehabilitation for pediatric brain injury. PMID- 12649691 TI - The importance of a comprehensive brain injury program. PMID- 12649692 TI - Functional capacity evaluations and the use of validity testing: what does the evidence tell us? PMID- 12649693 TI - Case management time tracking. PMID- 12649694 TI - [Today's rheumatological paleopathology]. PMID- 12649695 TI - Psoriatic arthritis as a mountain. AB - The concept of psoriatic arthritis (PsA) is not yet universally accepted. Indeed, few of the features said to be characteristic for PsA are pathognomonic, and a same patient can be classified as RA, SpA or PsA depending on the physician seen. The heterogeneity of PsA, the lack of significant differences in early-arthritis with and without psoriasis, and a pathogenesis somewhat different in PsA and psoriasis, also argue against the originality of PsA. Nevertheless, although PsA is possibly "just" a syndrome depending on the combination of numerous co-factors (perhaps shared with SpA and/or RA), its more achieved forms deserve to be segregated from other SpA and RA. Indeed, PsA best bridges the gap with SAPHO syndromes. Moreover, the profile of synovial cytokines seems somewhat different in PsA as compared to RA and SpA, and a special pattern of vascularisation has been confirmed by several teams which could account for the demonstrated link between trauma and some PsA onsets. Both the greater familial risk for PsA than for RA or psoriasis alone and the excessive paternal transmission of PsA strongly suggest a genetic background, although so far only MICA-A9 (expressed on gut epithelial cells) seems to be associated with susceptibility to PsA independently from psoriasis. To make further genetics studies informative, a careful selection of unequivocal cases of PsA is needed, which requires criteria selecting patients at the "top of the mountain" of PsA. One can expect that the sets of criteria proposed by McGonagle or Fournie could satisfy this wish. PMID- 12649696 TI - [Crystal-induced joint inflammation]. AB - The inflammatory response to microcrystals is one of the most powerful and intriguing examples of inflammation observable in man. Although many mechanisms of this reaction are well known, some aspects need to be further clarified, in particular those related to the self-limited nature of the process. Type and duration of the inflammatory reactions are mainly influenced by crystals characteristics, including shape and size, which, in turn may involve the crystal binding of several proteins, essential for the modulation of cellular responses. Cells most involved in the acute attacks are macrophage and neutrophils, which are responsible for the secretion of several important mediators of inflammation, such as prostaglandins and cytokines. These substances may in turn influence both intensity and duration of the acute attack. The proinflammatory effects of interleukin (IL)-1, IL-6, IL-8 and tumour necrosis factor (TNF)-alpha are counterbalanced by the anti-inflammatory activity of transforming growth factor (TGF)-beta, which may inhibit both the cell recruitment and cytokine synthesis. The role of TGF is crucial, not only by limiting acute inflammation but also by promoting formation and deposit of calcium crystals. However, the final effect of the balance between inflammatory and anti-inflammatory cytokines may also depend by other tissutal and cellular factors, not all of which are still completely understood. PMID- 12649697 TI - [Chromosome aberrations, valued as frequency of spontaneous micronuclei, in subjects with suspected presclerodermic Raynaud's phenomenon]. AB - OBJECTIVE: To evaluate the prevalence of spontaneous chromosome damage in cultured peripheral lymphocytes of subjects with suspected presclerodermic Raynaud's phenomenon (RP), by means of molecular cytogenetic analysis. METHODS: We studied 20 suspected presclerodermic RP, 20 idiopathic RP and 25 healthy subjects. As marker of chromosome alteration we used the micronucleus assay. All subjects were also classified as ANA-, ACA+ or Scl70+. To identify the mechanism of MN formation, a MN fluorescence in situ hybridisation (FISH) analysis using a pancentromeric DNA probe was also performed. RESULTS: Suspected presclerodermic RP subjects, showed significantly higher MN frequencies than idiopathic RP and controls (39+/-15.2 vs 10+/-2.1 and 9.8+/-3.5 respectively p<0.0001). Interestingly, subjects with idiopathic RP displayed MN frequency comparable to that of controls. Furthermore, ACA+ subjects showed the highest MN frequencies (44+/-8.1) as compared to subjects with different antibody pattern (26+/-7.1). CONCLUSIONS: Our results show the presence of higher levels of chromosomal damage in circulating lymphocytes of suspected presclerodermic RP. They also would suggest a key role of anti-centromere antibody in determining the observed cytogenetic anomalies. FISH analysis indicated that both aneuploidogenic and clastogenic events contribute to the formation of MN observed in suspected presclerodermic RP. PMID- 12649698 TI - [Evaluation of the peak bone mass by quantitative heel ultrasound in young women of the centre of Italy]. AB - OBJECTIVE: To measure the reference young adult mean values in healthy women of the centre of Italy by Quantitative heel UltraSound (QUS). METHODS: The study group was composed by 70 caucasian women: mean age was 25.4 years (Standard Deviation 4.7), mean weight was 58 Kg (SD 8.2), mean height was 166 cm (SD 5.8), mean BMI was 20.9 kg/m 2 (SD 2.5). Every subject was evaluated firstly with an original questionnaire to discover risk factors (like for example steroids consumption, recent fractures of the lower limb), then was measured by quantitative heel ultrasonometry Hologic Sahara. RESULTS: Mean extimated Bone Mineral Density (BMD) 0.588 g/cm 2 (SD 0.124) mean Quantitative Ultrasound Index (QUI) 105.0 (SD 19.6), mean Speed of Sound (SOS) 1564.2 m/s (SD 31.4), mean Broadband Ultrasound Attenuation (BUA) 84.8 dB/MHz (SD 17.4). No significant correlation was found between QUS parameters and anthropometric data. A correlation was found between every QUS parameters. No significant differences were found about QUI and extimated BMD, between our results and Hologic normative data for European women. CONCLUSIONS: It is very important to develop specific reference values for any measurement device and site of skeleton especially in the age of reaching the peak bone mass because the T score is then measured referring to these data. Usually the normative data are supplied by manufacturer and are based on large multicentric study. In our opinion it could be helpful to verify if these data are compatible with the population examined in every region. PMID- 12649699 TI - [Arthroscopic synovectomy in chronic inflammatory rheumatism: clinical and functional aspects]. AB - By now many authors regard arthroscopic synovectomy an integral part of therapeutic treatment of many rheumatic diseases with favourable results on post operating course and clinical picture in the long term. The pathologic synovial tissue during articular inflammatory rheumatism is well known to have a damaging effect responsible of early cartilage injury, as well as symptomatic action (e.g. articular stiffness, effusion, pain, functional limitation). Therefore to value the removal of such a tissue you should think of the secondary prevention of cartilage injury, besides the symptomatic point of view. Since 1996 we performed 190 arthroscopic synovectomy, the adopted criteria of judgement were: pain (spontaneous, during active and passive movements), effusion or swelling presence, articular range and cartilage state (evaluated during arthroscopy according to Outerbridg's classification). 70% of the cases showed good results and six years later the beginning of this activity we retain arthroscopic synovectomy as a valid help in articular inflammatory rheumatism treatment. PMID- 12649700 TI - [Safety of lornoxicam in G-6-PDH deficiency]. AB - OBJECTIVE: To assess the safety of lornoxicam in subjects with G-6-PDH deficiency. METHODS: Open controlled 2-week in vivo study on lornoxicam 8 mg bid in subjects with G-6-PDH deficiency suffering from rheumatic diseases. RESULTS: In 8 male patients with the Mediterranean form of G-6-PDH deficiency (mean age +/ SD, 54.3 years +/- 7.2) lornoxicam showed no influence on red blood cells (RBC) survival curve. The RBC half-life was the same before and after two weeks of treatment. CONCLUSIONS: Lornoxicam caused no RBC damage and evidenced favourable safety in subjects with G-6-PDH deficiency, suffering from rheumatic diseases. PMID- 12649701 TI - [Post-traumatic arthritis in a patient with Jadassohn's anetoderma]. AB - We describe the emergence of arthritis following a physical trauma, in a young man with clinical and histopathologic features of primary anetoderma (Jadassohn type) of 13 years' duration. Diagnosis of post-traumatic arthritis in a young patient with genetic predisposition was assumed. Indeed septic arthritis and other possible cause of arthritis were ruled out. PMID- 12649702 TI - [Hidden bone erosions]. AB - The aim of this pictorial essay was to demonstrate the diagnostic efficacy of high-resolution sonography in detecting bone erosions in a patient with rheumatoid arthritis. Standard X-Ray of the feet did not reveal clearly evident erosions. Ultrasonography was able to detect the presence of bone erosions of the metatarsal heads of both the first toes and of the V toe of the left foot. Because the appearance of bone erosions on radiographs of a patient with a recent onset arthritis indicates a poor prognosis, the possibility of demonstrating small hidden erosions at the level of the early targets of the disease is of relevant practical value. PMID- 12649703 TI - Frequency of pathology in a large natural sample from Natural Trap Cave with special remarks on erosive disease in the Pleistocene. AB - Population data are presented for erosive arthritis, osteoarthritis, diffuse idiopathic skeletal hyperostosis (DISH), joint eburnation and dental injury in a fauna from Natural Trap Cave, Wyoming, represented by over thirty thousand bones from twenty-four different species. Erosive arthritis is limited to the bovids, Bison, Ovis and Bootherium. Erosive arthritis is also present in bison from the late Pleistocene Twelve Mile Creek site in Kansas and from an early Holocene site in Wisconsin. The restriction of the known Pleistocene occurrences to bovids indicate the presence of a pathogen that predisposes bovids to erosive arthritis. The pathogen was identified as Mvcobacterium tuberculosis. Osteoarthritis and DISH are rare in the Natural Trap Cave, although Bison shows a relatively high occurrence of the former. Tooth breakage due to errors in bone manipulation was a problem for carnivores and one lion, Pantera atrox, was apparently reduced by joint disease to a scavenging lifestyle. PMID- 12649704 TI - Characterization of as-received, retrieved, and recycled stainless steel brackets. AB - AIM: The purpose of the present study was to investigate the composition and to assess the microhardness and structure of as received, retrieved, and recycled stainless steel brackets. MATERIALS AND METHODS: New, used, and recycled brand-, slot size-, and prescription-matched appliances were subjected to scanning electron microscopy (SEM), energy dispersive (EDS) electron probe microanalysis, metallographic analysis, and Vickers microhardness testing. Elemental analysis was performed on randomly selected bulk material base and wing areas. Basic metal content and microhardness results were statistically analyzed using a two-way ANOVA and the Tukey test, with treatment (as received, retrieved, and recycled) and bracket region (base, wing) serving as discriminating variables (alpha = 0.05). RESULTS: The results showed that there was no alteration in the bulk composition of the brackets among the three conditions. In contrast, differences were noted between bracket base and wing with respect to elemental composition among all groups, implying that the base and wings were manufactured from different alloys. The metallographic etching identified no difference in grain structure between the bracket base and wing components for all groups, regardless of treatment. Vickers microhardness demonstrated significant differences in hardness between base and wing for all groups and between recycled and retrieved states for the wing component. PMID- 12649705 TI - Transformation behavior, chemical composition, surface topography and bending properties of five selected 0.016" x 0.022" NiTi archwires. AB - PURPOSE: The aim of this study was to characterize five selected commercial NiTi archwires in terms of their transformation behavior, chemical composition, surface topography and mechanical properties (at temperatures of 22 degrees C, 37 degrees C and 60 degrees C). MATERIAL AND METHODS: The rectangular orthodontic archwires investigated were Neo Sentalloy F80 (GAC, Central Islip, NY, USA), 35 degrees C Thermo-Active Copper NiTi (A-Company/Ormco, Glendora, CA, USA), Rematitan "Lite" (Dentaurum, Pforzheim, Germany), Titanol SE S (Forestadent, Pforzheim, Germany) and Titanal (Lancer, San Marcos, CA, USA) in size 0.016" x 0.022". The chemical composition and surface topography were analyzed by energy dispersive X-ray spectroscopy using an analytical scanning electron microscope (XL30, EDAX SUTW Saphire Detector; Philips, Eindhoven, Netherlands). The transition temperatures were measured by means of differential scanning calorimetry (DSC; Perkin-Elmer Pyris 1, Perkin-Elmer, Fremont, CA, USA) in a range of - 80 degrees C to + 80 degrees C. The mechanical properties and their dependence on temperature were determined by means of 3-point bending tests. The binary archwire materials were characterized by a two-phase structure (NiTi matrix and Ni3Ti4 precipitates). RESULTS: The SEM analyses revealed abradant residues in virtually all archwires, while DSC revealed complex transformation properties. In addition to the martensitic and austenitic transformations, an R phase transformation was also detected. The bending tests showed pronounced loading and unloading plateaus. The martensitic archwires (Neo Sentalloy F80, 35 degrees C Thermo-Active Copper NiTi) were found to have a lower strength than the martensitic-austenitic (Rematitan "Lite") and the austenitic archwires (Titanol SE S, Titanal). With increasing temperature (in the range from 22 degrees C to 60 degrees C) a linear rise in the plateau forces was recorded. CONCLUSIONS: When assessing the quality of archwires, account should be taken of the surface quality, as it is this that determines corrosion resistance, biocompatibility and friction characteristics. The mechanical properties depend on the initial state; moderate plateau forces and plateau moments can only be achieved with martensitic archwires. In contrast to conventional steel alloys, the strength characteristics are heavily dependent on temperature and need to be known if NiTi archwires are to be used to optimal effect. In addition, the superelastic plateau is used only partially, if at all, when minimum leveling is required. PMID- 12649706 TI - Determination of the mechanical properties of the periodontal ligament in a uniaxial tensional experiment. AB - BACKGROUND: The periodontal ligament is a soft connective tissue which joins the tooth root to the alveolus and thus provides for anchorage of the tooth in the alveolar bone. Due to its composition of elastic and viscous components, this tissue displays viscoelastic material properties. In a previous study [4], in vitro experiments revealed typical viscoelastic material properties of the periodontal ligament in samples from pig mandibles. These properties included force relaxation, hysteresis, and dependence on loading history. MATERIAL AND METHODS: Based on those experiments, a dependence of tooth displacement on loading velocity was registered in the present study and the stress-strain behavior of the periodontal ligament was examined until the tissue ruptured. For this purpose, segments of the periodontal ligament taken from anterior teeth from the pig mandible were tested in a purpose-developed clamping fixture in a uniaxial tensional experiment. RESULT: It was found that the initial phase of the stress-strain curve in particular was dependent on loading velocity and that the shape of the hysteresis curve was subject to a variation in loading velocity. The stress-strain behavior of the periodontal ligament was characterized, divided into several phases, and the elastic modulus of the initial and the linear phase of the curve was determined at different loading velocities. CONCLUSION: Knowledge of the material properties of the periodontal ligament is fundamental to an understanding of orthodontic tooth movement and thus to selection of an optimal force system for orthodontic treatment. PMID- 12649707 TI - Dental anomalies as a microsymptom of palatal canine displacement. AB - AIM: A retrospective study was performed to investigate whether palatal canine displacement is associated with other dental features permitting early clinical diagnosis of the eruption disturbance. PATIENTS AND METHODS: The study was based on the complete records of 235/8556 patients at the Department of Orthodontics, School of Dental Medicine, University of Freiburg i.Br. (mean age = 14.11 years) with at least one palatally displaced permanent canine. These patients were examined for ten different morphologic parameters, e.g. impaction and congenital absence of further teeth, hypoplastic, peg-shaped, rotated and congenitally missing upper lateral incisors, supernumerary teeth, Angle classification, and cover-bite (= "Deckbiss"). The data were compared with those of a control sample of the same size with physiologic upper canine eruption (mean-age = 10.0 years). Furthermore, in a right/left comparison the local influence of anomalies of the upper lateral incisors on palatal canine displacement was evaluated by multiple linear regression analysis. RESULTS: The statistical analyses revealed that the risk of palatal canine displacement was significantly higher in patients with hypoplasia, peg shape or congenital aplasia of upper lateral incisors, further impacted and congenitally missing teeth and cover-bite. The intergroup differences in terms of gender, rotation of upper lateral incisors and Angle classification were not statistically significant. The right/left comparison revealed a significantly higher risk of palatal canine displacement in association with an adjacent hypoplastic or peg-shaped lateral incisor and with aplasia of the contralateral upper incisor. CONCLUSION: The clinical significance of the study is that the occurrence of palatally displaced canines is often closely linked with other dental anomalies. In this context, anomalies of upper lateral incisors, aplasia or impaction of further teeth, and the anomaly of cover bite may serve as indicators of palatal canine displacement. A retarded development of the upper lateral incisor seems to be more disturbing for physiologic canine eruption than aplasia. In patients exhibiting the stated microsymptoms, close clinical follow-up of the maxillary permanent canine eruption during the late exfoliation period is strongly recommended. PMID- 12649708 TI - Interocclusal clearance during speech and in mandibular rest position. A comparison between different measuring methods. AB - BACKGROUND AND AIM: The interocclusal clearance during speech and in mandibular rest position shows an interindividual variation and influences the stability of prosthodontic reconstruction or orthodontic therapy, especially in patients with deep bite or cover-bite. Exact determination of the vertical dimension, not always a simple matter in practice, is controversially discussed with respect to the methodology. The aim of the present investigation was to compare three methods for determining interocclusal clearance during speech and in mandibular rest position. PATIENTS AND METHODS: Extraoral manual registration and electronic registration were used to evaluate 33 malocclusion-free patients. The measurements were repeated to determine the reproducibility. For disturbance-free objectification, the freeway space, i.e. the interocclusal clearance in mandibular rest position, was also recorded cephalometrically. RESULTS: None of these registration methods displayed clear-cut superiority to the others. Electronic registration is very time-consuming and should be confined to special cases. Sources of error in manual registration are increased movement of the measuring points marked on the skin during mandibular movement and the free handling of dividers. CONCLUSIONS: This method can be used only in connection with mm. consonants. Speaking the word "Ohio" yielded excessively high values in all methods, so that this word has to be rejected as a speech sample. Cephalometric registration produced values with slight interindividual variations. In practice, however, this method is unsuitable for use with orthodontic patients because of the additional radiation exposure involved in producing an additional lateral cephalogram. For good reproducibility, practicing or frequent repeating of the measuring method prior to definitive measuring is essential. PMID- 12649709 TI - Evaluation of the parameters underlying the decision by adult patients to opt for lingual therapy: an international comparison. AB - BACKGROUND: The present study is the first instance of patients having been questioned on the criteria applied in their decision for therapy with lingual brackets and on the sources of information available to them within the framework of an international comparison prior to the start of therapy by means of a standardized questionnaire. PATIENTS AND METHOD: 68 patients (19 male, 49 female; mean age 32.4 years, range 19.8 to 60.0 years, SD 9.7) were interviewed. RESULTS: Therapy with lingual brackets was more important to the patients in the present study for professional than for private reasons: The further the patients were from the typical "braces age", the more important was treatment with an invisible appliance from the professional standpoint. The patients' educational level had no influence on the decision for a fixed lingual appliance. There was no significant difference between the German and the French patients in the decision making criteria of adult patients and the sources of information available to them in the context of the lingual technique. CONCLUSION: The information available through the media on the lingual technique is inadequate in both countries. PMID- 12649710 TI - Prospective evaluation of the epidemiology, microbiology, and outcome of bloodstream infections in hematologic patients in a single cancer center. AB - Bloodstream infections (BSIs) have an important impact on the outcome of cancer patients. A prospective cohort study was undertaken at a referral cancer center in order to describe the clinical and microbiological characteristics of patients with hematologic malignancies and BSIs and to identify independent predictors associated with mortality. The study enrolled 110 consecutive BSI episodes during an 18-month period. Patients were monitored for 30 days after the last positive blood culture. There were 10.24 BSI episodes per 1,000 patient-days. The median age of the patients was 25 years. Most patients had acute leukemia ( n=72). The origin of the BSI was unknown in 43.6% of the episodes and was associated with known sites in 32.7%. There were 58 concomitant infectious sites (lungs, 43%, and soft tissue, 22.4%) and 195 noninfectious comorbid factors (poor performance status, 30.2%; undernourishment, 14.3%). The median neutrophil count was 215 cells/mm(3). Indwelling catheters were present in 70% of the episodes. The majority of isolates obtained within the first 48 h of the BSI episode (61%) were gram-negative rods. Overall mortality was 24.5%. Multivariate analysis using logistic regression showed relapsed leukemia, poor performance status, recent weight loss, and ventilatory failure requiring ventilatory support as independent predictors of mortality. Hematologic cancer patients with BSIs should be regarded as a distinct group of patients at high risk of death. The knowledge of variables amenable to intervention would help diminish or prevent serious medical complications. PMID- 12649711 TI - Randomised double-blind comparison of oral gatifloxacin and co-amoxiclav for acute exacerbation of chronic Bronchitis. AB - Antimicrobial therapy can have a significant impact in the treatment of acute infectious exacerbations in patients with chronic bronchitis, in whom repeated episodes are common. The aim of this randomised, double-blind, double-dummy, parallel group study was to compare the efficacy and safety of oral gatifloxacin (200 and 400 mg once daily) administered for 5 days with co-amoxiclav (500 mg amoxicillin/125 mg clavulanic acid t.i.d.) administered for 10 days in 414 adult patients with acute exacerbation of chronic bronchitis. Overall clinical response rates (cure plus improvement) were 86.2%, 79.4% and 81.7% in the gatifloxacin 200 mg, gatifloxacin 400 mg and co-amoxiclav groups, respectively, and the equivalence hypothesis used for statistical analysis showed equivalent efficacy for both gatifloxacin 200 and 400 mg compared to co-amoxiclav. The same was true for rates of bacterial response, with eradication or presumed eradication of causative pathogens achieved in 87.5%, 87.3% and 79.1% of cases in the gatifloxacin 200 mg, gatifloxacin 400 mg and co-amoxiclav groups, respectively. All treatments were well tolerated, with the nature and frequency of treatment related adverse events similar in all groups. The results of the study show that gatifloxacin is a safe and effective agent for the treatment of patients with chronic bronchitis experiencing an acute infectious exacerbation. PMID- 12649713 TI - Use of time-trend analysis in the design of empirical antimicrobial treatment of urinary tract infection. AB - Selection of empirical treatment of hospitalized patients with urinary tract infection (UTI) is usually based on the results of urine culture as obtained from the local microbiology laboratory. In order to improve the precision and reliability of traditional methods, we analyzed temporal changes in the results of urine culture and antibiograms and stratified the results by inpatient department and the presence/absence of an indwelling catheter. The database consisted of urine cultures obtained during the first 3 months of each year over a 10-year period between 1991 and 2000. Only urine samples that grew a single organism at a concentration of >10(5) cfu were included in the analysis. Trend statistical tools, readily available but thus far not used for microbiological analyses, were applied to assess the decay in activity of individual antibiotic agents over time and to calculate susceptibility rates of organisms in subsets of urine samples. Organisms, antimicrobial susceptibility rates and the degree of decay in antimicrobial susceptibility rates varied significantly according to the location of the patient in the hospital and the presence of an indwelling catheter. Stratified trend analysis is a useful tool that can be helpful in designing and adapting clinical guidelines for the selection of appropriate empirical antibiotic treatment for the individual patient with urinary tract infection. PMID- 12649712 TI - Factors associated with complications and mortality in adult patients hospitalized for infectious cellulitis. AB - The aim of this study was to analyze medical outcomes, including risks for complications and mortality, in 332 adult patients hospitalized for cellulitis. The infection was documented microbiologically in 128 cases (39%). Staphylococcus aureus (46 cases) and Streptococcus pyogenes (22 cases) were the most frequent causative pathogens. Overall, 63 patients (19%) were discharged early (< or =4 days) and 166 patients (50%) were hospitalized for more than 4 days without developing any complications. One hundred three patients (31%) had one or more complications or died. Of these, 78 required surgical debridement, 10 required plastic surgery, 7 underwent amputation, and 15 had shock on presentation. When comparing the three study groups (patients discharged early, patients hospitalized for < or =4 days without complications, and patients who developed 1 or more complication or who died), patients who were discharged early (low risk) were more frequently female and were less likely to have multiple comorbid conditions, hypoalbuminemia, renal insufficiency, and/or cutaneous necrosis at presentation. Overall mortality (<30 days) was 5% (16/332 patients). Factors associated with death were male sex, presence of multiple comorbid conditions, congestive heart failure, morbid obesity, hypoalbuminemia, renal insufficiency, shock, and Pseudomonas aeruginosa cellulitis. These findings can be used to stratify patients with acute cellulitis according to risks for complications and mortality and may be helpful when deciding the most appropriate means of care, i.e. outpatient treatment or hospitalization. PMID- 12649714 TI - Prevalence of Borrelia burgdorferi species and identification of Borrelia valaisiana in questing Ixodes ricinus in the Lyon region of France as determined by polymerase chain reaction-restriction fragment length polymorphism. AB - Many cases of Lyme borreliosis have been reported over the years in the region of Lyon, France. The identification and prevalence of Borrelia burgdorferi sensu lato in Ixodes ricinus were investigated by polymerase chain reaction (PCR) of the flagellin gene and restriction fragment length polymorphism (RFLP) analysis. Questing Ixodes ricinus larvae, nymphs and adults were collected by the flagging method from deciduous forests in four areas in the Lyon region of France between October 1994 and September 1995 and in June 1998. The overall prevalence of Borrelia burgdorferi sensu lato was 13.2% (91/688). No significant differences in prevalence were observed between the different stages and sex of the ixodids or between collection areas. The majority of infections were simple infections (82.4%; 75/91), most of which were due to Borrelia afzelii (41.4%), while coinfections (12.1%) were predominantly (54.5%) a combination of Borrelia valaisiana and Borrelia garinii. No tick was infected with more than two borrelial species, nor was Borrelia lusitaniae identified. The Borrelia valaisiana species was detected for the first time in France, confirming its widespread presence in Europe. This study confirms that the surroundings of Lyon are risk areas for contracting Lyme disease and that no particular clinical manifestations predominate due to the heterogeneous distribution of Borrelia genospecies. Moreover, the polymerase chain reaction restriction fragment length polymorphism analysis is a rapid and easy method for genotyping of Borrelia species. PMID- 12649715 TI - Evaluation of a commercial IgG/IgM Western blot assay for early postnatal diagnosis of congenital toxoplasmosis. AB - The aim of this study was to evaluate a commercial Western blot IgG/IgM assay for use in the early serological diagnosis of congenital toxoplasmosis. This assay compares the immunological profile of mother and infant and allows differentiation between passive transmitted maternal antibodies and newly synthesized antibodies of the infant within the first 3 months of life. Over a 6 year period (1995-2001), the sera from 169 mothers and their 175 offspring (6 had twins) were examined for specific anti- Toxoplasma gondii IgG, IgM and IgA antibodies with an enzyme-linked immunosorbent assay or an immunosorbent agglutination assay. All mothers had primary Toxoplasma infection during pregnancy. Serological and clinical follow-up of the infants during the first year of life confirmed 36 cases of congenital toxoplasmosis. In 139 cases, infection could be ruled out. Three hundred fifty-one paired samples from 175 mother-child pairs were tested retrospectively for IgG and IgM patterns by Toxoplasma Western blot IgG/IgM (LDBIO Diagnostics, France). The results of conventional serological analysis (immunosorbent agglutination assay or enzyme linked immunosorbent assay) to detect IgM or IgA were compared with the results of the Toxoplasma Western blot IgG/IgM on samples obtained within the first 3 months of life. The performance of the combination of the two methods was also assessed. At birth, the sensitivity values of conventional serological analysis and the Toxoplasma Western blot were 52% and 67%, with specificity values being 99% and 96%, respectively. Combination of the Western blot and conventional serological analysis increased the sensitivity at birth to 78% and within the first 3 months of life to 85%. Overall, the combination of both methods detected 94% of congenital infections. Therefore, this commercial Western blot represents a useful tool for early postnatal diagnosis of congenital toxoplasmosis. PMID- 12649716 TI - Cellular immunity to Toxoplasma gondii in congenitally infected newborns and immunocompetent infected hosts. AB - The aim of this study was to determine the frequency of anergy to Toxoplasma gondii in congenitally infected newborns and immunocompetent infected individuals. Specific anergy to Toxoplasma has been reported previously, especially in cases of congenital toxoplasmosis. Whole blood from 592 immunocompetent patients with suspected toxoplasmosis was cultured in the presence of soluble Toxoplasma antigen for 7 days. Activated T lymphocytes were detected by flow cytometry. In patients over 1 year of age, the percentage of soluble Toxoplasma antigen-stimulated T cells expressing the interleukin-2 receptor CD25 was higher in infected patients than in uninfected subjects (40.0+/ 18.3% vs. 1.8+/-2.0%, P<0.0001). No differences were detected between seroconverters, i.e. those with recent rises in IgM and IgG antibodies, and those with acquired or congenital toxoplasmosis. Similar results were observed when congenitally infected ( n=38) and uninfected ( n=89) children under 1 year of age were compared (17.6+/-9.2% vs. 3.0+/-4.9%, P<0.0001). The sensitivity and specificity values of CD25 detection for diagnosis of congenital toxoplasmosis in infants were 95% and 89%, respectively, at a threshold value of 7% above control culture. The results show that specific cellular immunity is detectable in virtually all Toxoplasma-infected patients, including newborns. Detection of CD25 constitutes a simple, sensitive and specific test for diagnosis of congenital toxoplasmosis. PMID- 12649717 TI - Efficacy and safety of sequential amoxicillin-clavulanate in the treatment of anaerobic lung infections. AB - In order to assess the efficacy and safety of amoxicillin-clavulanate for the treatment of anaerobic lung infection, 40 patients with lung abscess or necrotizing pneumonia were given sequential amoxicillin-clavulanate therapy. All patients received intravenous amoxicillin-clavulanate (2 g/200 mg/8 h), which was switched to oral form (1 g/125 mg/8 h) after clinical improvement. Mean duration of antibiotic therapy was 43.5 days. Microbiological documentation was obtained in 53% of cases. All but 1 of the 48 microorganisms isolated were susceptible to amoxicillin-clavulanate. The drug was well tolerated by the patients and no severe adverse effects were observed. At the end of treatment all patients were considered cured. The 35 patients assessed at long-term follow-up visit remained disease-free. PMID- 12649718 TI - Analysis of the case fatality rate of tetanus among adults in a tertiary hospital in Turkey. AB - The aim of this study was to present the epidemiological and clinical characteristics of tetanus as related to case fatality in 43 adult patients hospitalized between 1990 and 2000 in Turkey's largest state tertiary hospital. All patients were > or =15 years of age, with the mean age being 45 years. Sixty five percent of the patients were male. Most patients (77%) were from rural Anatolia. The most common clinical manifestations were trismus (100%), abdominal rigidity (93%), dysphagia (81%), and risus sardonicus (72%). The overall case fatality rate was 58%. In a multivariable logistic regression model an incubation period of < or =8 days versus >8 days was a significant predictor of case fatality (odds ratio, 4.8; P=0.044; 95%CI, 1.04-22.26), as was each day of delay between onset of symptoms and admission to the hospital (odds ratio, 7.4; P=0.035; 95%CI, 1.15-48.28). Although the case fatality rate of tetanus in our clinic has declined in the last 5 years, the overall rate was high. This suggests a need for adult vaccination programs and/or a plan to educate the public to recognize symptoms early and then rapidly seek medical care. PMID- 12649719 TI - Three ECHOvirus serotypes responsible for outbreak of aseptic meningitis in Rhone Alpes region, France. AB - During the year 2000 in the Rhone-Alpes region of France, 559 cases of aseptic meningitis due to enterovirus infection were recorded (approximate incidence, 10 cases per 100,000 population compared with a mean of 0.3 cases during endemic years in this region; P<0.001). Cerebrospinal fluid samples were collected from all 559 patients and processed for enterovirus RNA detection using a reverse transcriptase polymerase chain reaction assay only. In addition to the cerebrospinal fluid samples, 40 stool and 76 throat samples were collected from 116 patients (20.7% of cases); the following three enterovirus serotypes were isolated from the stool and throat samples only: ECHOvirus 13 (48/116; 41.3%), ECHOvirus 30 (44/116;37.9%) and ECHOvirus 6 (24/116, 20.7%). This is the first report that demonstrates the involvement of three different ECHOvirus serotypes, particularly ECHOvirus 13, in an outbreak of aseptic meningitis, in the French Rhone-Alpes region. PMID- 12649720 TI - Influence of hepatitis C virus coinfection on failure of HIV-infected patients receiving highly active antiretroviral therapy to achieve normal serum beta2microglobulin levels. AB - The influence of hepatitis C virus (HCV) coinfection on the kinetics of serum beta2microglobulin levels in HIV-infected patients undergoing highly active antiretroviral therapy was analyzed. After 96 weeks of therapy, beta2microglobulin levels declined significantly both in 24 HCV seronegative and 16 HCV-infected individuals. Throughout the follow-up period, HCV-infected patients showed higher beta2microglobulin values than non-HCV-infected patients. These results show that HCV infection precludes immune deactivation in HIV infected patients undergoing highly active antiretroviral therapy. PMID- 12649721 TI - Myocarditis associated with reovirus infection. PMID- 12649722 TI - Laboratory detection and investigation of reduced susceptibility to vancomycin in oxacillin-resistant Staphylococcus aureus. PMID- 12649723 TI - Comparative morphological study of the lingual papillae and their connective tissue cores of the koala. AB - The stereo structure of each lingual papilla of the koala has a similar structure to that of various other animal species: the koala has a lingual prominence (intermolar prominence) with larger filiform papillae. (A lingual prominence is a characteristic in herbivorous animals.) The external form and connective tissue core (CTC) of the filiform papillae of koalas consist of one large main process and several smaller accessory processes. (These are similar to carnivorous animals.) Fungiform CTC have a thick dome-like structure, with several taste buds on the top. There are three vallate papillae: one central midline and two laterally located vallate papillae. The central vallate papilla has a posterior pouch lined with ciliated and non-ciliated epithelial cells. Long conical papillae are distributed in the posterior lateral area where foliate papillae are distributed in many other animal species. (Finger-like papillae are seen in dog and cat instead of foliate papillae.) It may be suggested that the tongue of the koala evolved in a special environment in Australia. Even though it has still retained special features similar to those of carnivorous cats and dogs it has evolved to resemble the tongues of herbivorous animals. PMID- 12649724 TI - The genus Acipenser as a model for vertebrate urogenital development: the mullerian duct. AB - The development of the mullerian duct was studied in a total of 85 specimens of Acipenser ruthenus and Acipenser baeri in the period from 7 days after hatching through 5 years of age. Normal histology on serial sections, transmission and scanning electron microscopy and bromodeoxyuridine immunohistochemistry were applied. In Acipenser, the primary set of opisthonephric nephrons possess short nephrostomial tubules with well-developed nephrostomes. Proliferating cells from the lateral side of the slightly protruding nephrostomial lips spread out over the ceolomic surface, replace here the flat mesothelium of lateral plate origin and establish the infundibular field, consisting of cuboidal or columnar cells. At about 28 days after hatching, the primordium of the mullerian infundibulum becomes visible in the form of a pocket-like invagination within the infundibular field. This invagination is found coexisting with and located laterally to the line of intact nephrostomes. The mullerian infundibulum, therefore, does not represent the homologon of a nephrostome itself, but must be regarded as a separate and secondary structure. The mullerian duct proper has its origin in cells from the bottom of the infundibular pocket. These cells grow as a tubule with a solid tip in the caudal direction, paralleling the wolffian duct, but without a contribution of cells from the latter. In Acipenser, a mullerian duct is present also in the adult male. In males as in females, the caudal extremity of the mullerian duct generally divides into two to three smaller terminals which end in the wolffian duct at different levels, but always cranial to the urogenital sinus. In most indifferent animals and in all males of this study, the tips of the mullerian terminals are closed and covered by a thin layer of wolffian epithelium. In adult females, the mullerian ducts end with open terminals. In both sexes, the adult mullerian duct is lined by a pseudostratified columnar epithelium, consisting of ciliated, non-ciliated and basal free cells. PMID- 12649725 TI - Ultrastructure of the central subnucleus of the nucleus tractus solitarii and the esophageal afferent terminals in the rat. AB - The central subnucleus of the nucleus tractus solitarii (ceNTS) receives afferent projections from the esophageal wall and projects to the nucleus ambiguus, thus serving as a relay nucleus for peristalsis of the esophagus. Here we examine the synaptic organization of the ceNTS, and its esophageal afferents by using transganglionic anterograde transport of cholera toxin-conjugated horseradish peroxidase (CT-HRP). When CT-HRP was injected into the subdiaphragmatic esophagus, many anterogradely labeled terminals were found only in the ceNTS. The ceNTS was composed of round or oval-shaped, small neurons (14.7x8.7 micro m) containing sparse organelles and an irregularly shaped nucleus. The average number of axosomatic terminals was only 1.3 per section cut through the nucleolus. Most of them (92%) contained round vesicles and formed asymmetric synaptic contacts (Gray's type I), and a few (8%) contained pleomorphic vesicles and formed symmetric synaptic contacts (Gray's type II). All anterogradely labeled terminals contacted dendrites but not the neuronal somata. The labeled terminals were large (2.55+/-0.07 micro m) and exclusively Gray's type I. More than half of them (60%) contacted small dendrites (less than 1 micro m in diameter), and contained dense-cored vesicles. More than 40% of the labeled terminals contacted two to four dendrites, thus forming a synaptic glomerulus. Sometimes a labeled terminal that contacted an unlabeled terminal by an adherent junction was found within the glomerulus. The large terminals and these complex synaptic relations appeared to characterize the esophageal afferent projections in the ceNTS. PMID- 12649726 TI - The relationship between limb muscle and endothelial cells migrating from single somite. AB - Somites contribute myogenic and endothelial precursor cells to the limb bud. Transplantations of single somites have shown the pattern of muscle cell distribution from individual somites to individual limb muscles. However, the pattern of the endothelial cell distribution from individual somites to the limb has not been characterized. We have mapped quail muscle and endothelial cell distribution in the distal part of the chick limb after single somite transplantation to determine if there is a spatial relationship between muscle and endothelial cells originating from the same somite. Single brachial somites from quail donor embryos were transplanted into chick embryos, and, following incubation, serial sections were stained with a quail-endothelial cell-specific monoclonal antibody (QH-1), an anti-quail antibody (QCPN) and an anti-desmin antibody to distinguish the quail endothelial and muscle cells from chick cells. Our results show that transplants of somite 16-21 each gave rise to quail endothelial cells in the wing. The anterioposterior position of the blood vessels formed by somitic endothelial cells corresponded to the craniocaudal position of the somite from which they have originated. Endothelial cells were located not only in the peri- and endomysium but also in the subcutaneous, intermuscular, perineural and periost tissues. There was no strict correlation between the distribution of muscle and endothelial cell from a single transplanted somite. Blood vessels formed by grafted quail endothelial cells could invade the muscle that did not contain any quail muscle cells, and conversely a muscle composed of numerous quail muscle cells was lacking any endothelial cells of quail origin. Furthermore, a chimeric limb with very little quail muscle cells was found to contain numerous quail endothelial cells and vice versa. These results suggest that muscle and endothelial cells derived from the same somite migrate on different routes in the developing limb bud. PMID- 12649727 TI - Differential distribution of melatonin receptors in the pituitary gland of Xenopus laevis. AB - A major target site for melatonin action is thought to be the pituitary gland. We have detected differential expression and co-localization of the Mel(1a) and Mel(1c) receptors in cells of the Xenopus laevis pituitary gland. Sections of Xenopus pituitary glands were labeled with Mel(1a) and/or Mel(1c) antibodies, in combination with antibodies to arginine vasotocin (AVT), alpha-melanocyte stimulating hormone (alpha-MSH), prolactin (PRL), and luteinizing hormone (LH). Mel(1a) immunoreactivity was localized to cells of the pars intermedia and to elements within the pars nervosa. Mel(1c) immunoreactivity was also localized to the pars nervosa, and significant labeling was also observed in discrete clusters of cells in the pars distalis. Mel(1a) was absent from the pars distalis, while Mel(1c) was absent from the pars intermedia. Mel(1a) and Mel(1c) were co localized in the pars nervosa. AVT was present in the pars nervosa, and appeared to be localized to the cell clusters of the pars distalis in which the Mel(1c) receptor was localized. alpha-MSH co-localized with the Mel(1a) receptor in the pars intermedia. LH appeared to localize to many of the cells in the pars distalis, with the notable exception of the Mel(1c) receptor-positive clusters of cells. PRL did not appear to co-localize with either melatonin receptor. The pattern of differential expression of the Mel(1a) and Mel(1c) receptors suggests that the receptors specifically mediate the cellular response to melatonin binding in the specific cell populations. PMID- 12649728 TI - Spontaneous remission of edema and regranulation of goblet cells in rat tracheae after capsaicin-induced acute inflammation. AB - Previous studies have investigated the short-term effect of capsaicin on edema formation and goblet-cell secretion in the trachea. The present study sought to investigate the long-term effect of a high dose of capsaicin (90 micro g/ml/kg), administered intravenously, on changes in the formation of endothelial gaps among venular endothelial cells, mucosal tissue edema and the secretory activity of goblet cells, including the number and size of goblet cells, and the mucus score and secretory ratio of goblet-cell mucus secretion in the trachea of rats. The tracheal whole mounts with silver staining, those stained with chloroacetate esterase reagent and Alcian blue and tracheal tissue sections stained with Alcian blue and periodic acid-Schiff reagent were used for evaluation. Formation of endothelial gaps occurred a few min after administration of capsaicin, and gaps almost closed within 12 min after capsaicin injection. Five min after capsaicin, the leaky blood vessels were numerous and the subepithelial edema ratio (% of length of edema along the inner circumference of tracheal cross section) was found to be 57.8+/-3.0% ( n=6). The number of Alcian blue-positive goblet cells (1,090+/-220 per mm(2) of mucosal surface) was reduced to half the number of goblet cells in the vehicle-treated rats (2,200+/-230). The mucus score of goblet cell secretion was not changed. The secretory ratio was greatly increased. One day after capsaicin, the edema ratio remained large and the number of Alcian blue positive goblet cells was also small. The mucus score was also not changed. The secretory ratio was still large. On day 3, the edema ratio remained large, but the number of Alcian blue-positive goblet cells was increased to the level of the controls. The mucus score and secretory ratio returned to the control level. On day 5, the edema ratio was greatly decreased, but it was still significantly larger than that of the controls. The mucus score and secretory ratio remained at the baseline level. Seven days after capsaicin, the edema ratio was similar to the controls. The number of goblet cells was even larger than controls. It is concluded that capsaicin-induced acute inflammation in the rat trachea involves formation of endothelial gaps, extensive plasma extravasation and edema formation, and depletion of goblet-cell secretory granules. Spontaneous gradual remission of edema was accompanied by regranulation of goblet cells with gradual mucogenesis for several days. PMID- 12649729 TI - Brain-derived neurotrophic factor in higher vertebrate pancreas: immunolocalization in glucagon cells. AB - Brain-derived neurotrophic factor (BDNF) is a growth factor that belongs to the group of neurotrophins. Its amino acid sequences are well conserved during vertebrate phylogenesis. Pancreatic tissue has recently been reported to be one of the physiological sources of BDNF in humans and mice. In this study we investigated the presence and localization of BDNF immunoreactivity (IR) in the pancreas of three species of higher vertebrates: mouse, duck and lizard. BDNF IR was present in the islets and in single cells scattered in the exocrine parenchyma of all three species examined. Using double staining, BDNF IR was seen to be colocalized with glucagon IR in all the species studied. There was a total overlap of BDNF and glucagon IR in duck and lizard pancreas, and partial overlap in mouse pancreas. Our findings suggest that, as well as the primary structure, the presence and pattern of distribution of BDNF in higher vertebrates is also well conserved. Moreover, the abundance of BDNF IR in the pancreas of the species studied leads us to the suggestion that these neurotrophins could regulate the function of pancreatic innervation and/or act on pancreatic cells in a paracrine/autocrine fashion. PMID- 12649730 TI - Ontogeny of the VIP system in the gastro-intestinal tract of the Axolotl, Ambystoma mexicanum: successive appearance of co-existing PACAP and NOS. AB - Evidence for the presence and potential co-existence of vasoactive intestinal polypeptide (VIP), pituitary adenylate cyclase-activating polypeptide (PACAP) and nitric oxide synthase (NOS) in gastro-intestinal endocrine cells and/or nerve fibers is conflicting and very few results exist on development. This immunofluorescence study aims to clarify the appearance and localization of VIP, PACAP and NOS in the gastro-intestinal tract of the Axolotl, Ambystoma mexicanum, during ontogeny. VIP-immunoreactivity appeared in nerve fibers as early as on day 3 after hatching likely indicating a particular role, such as a trophic action, of VIP in very early development. PACAP-immunoreactivity was observed 3 days later within the VIP-immunoreactive (-IR) fibers. From this time on, VIP- and PACAP-immunoreactivity exhibited complete co-existence. VIP/PACAP-IR fibers were found throughout the gastro-intestinal tract. They were most prominent in the myenteric plexus and the muscle layers and less frequent in the submucosa. NOS immunoreactivity appeared as late as at the 1st (64 days) juvenile stage in a subpopulation of the VIP/PACAP-IR fibers that contacted submucosal arteries. We found only very few VIP/PACAP-IR perikarya, indicating that part of the VIP/PACAP IR fibers is of extrinsic origin. On day 12 and in the 1st and 2nd (104 days) juvenile stage, infrequent PACAP-IR entero-endocrine cells were noted, while neither VIP- nor NOS-immunoreactivity occurred in endocrine cells at any stage of development. The complete coexistence of neuronal PACAP- and VIP immunoreactivities and their very early appearance in ontogeny may suggest important and coordinated roles of both peptides in the control of Axolotl gastro intestinal activity, while the VIP/ PACAP/NOS-IR fibers may be involved in the regulation of submucosal blood flow. PMID- 12649731 TI - The myth of ventrally emigrating neural tube (VENT) cells and their contribution to the developing cardiovascular system. AB - The cardiac neural crest cells are a group of cells that emigrate from the dorsal side of the neural tube during a specific time window and contribute to the pharyngeal arch arteries and the aorticopulmonary septum of the heart. Recent publications have suggested that another group of cells emigrating from the ventral side of the neural tube also contributes to the developing cardiovascular system. The first aim of our study was to define the specific time window of cardiac neural crest cell migration by injecting a retrovirus containing a lacZ reporter gene into a chick embryo at different stages during development. The second aim was to study the contribution of the supposed ventrally emigrating neural tube cells to the cardiovascular system using three approaches. One approach was to inject a lacZ retrovirus into the lumen of the chick hindbrain. Secondly, we injected the retrovirus into the neural tube at the position of the 10-12 somite pair. Finally, we used the chimera technique in which we transplanted a quail neural tube segment into a chick embryo. Cardiac neural crest cells were shown to emigrate from the dorsal side of the neural tube between HH9 and HH13(-). The HH13(+) neural tube has ceased to produce cardiac neural crest cells between the level of the otic placode and the fourth pair of somites. Retroviral injection directly into the chick hindbrain at HH14 resulted in 50% of the embryos with minimal labeling of the hindbrain and intense labeling of the adjacent mesenchyme, suggesting that virus was spilled. This implies that this technique is not useful for confirming the existence of ventrally emigrating cells. Both retroviral injections into the neural tube lumen at HH14 at the position of the 10-12 somite pair and the chimeras showed no signs of ventrally emigrating neural tube cells. We conclude that there is no contribution of ventral neural tube cells to the developing cardiovascular system. PMID- 12649732 TI - Evidence for the capacitation-associated membrane priming of mouse spermatozoa. AB - An important feature of male fertility is the physiological priming of mammalian spermatozoa by a multifaceted process referred to as capacitation. It is a prerequisite event before spermatozoa can bind to the egg's extracellular coat, the zona pellucida, and undergo a signal transduction cascade. The net result is the fusion of the plasma membrane (PM) and underlying outer acrosomal membrane at multiple sites and the release of acrosomal contents (i.e., glycohydrolases, proteinases, etc.) at the site of sperm-zona binding. In this study, we have used an indirect immunofluorescence (IIF) assay and other staining approaches to examine capacitation-associated membrane priming of mouse spermatozoa. For IIF studies, we used affinity-purified antibodies against two glycohydrolases that cross-reacted with the acrosomal enzymes only when the uncapacitated spermatozoa were permeabilized. Incubation of spermatozoa in a medium that favors in vitro capacitation induced membrane priming that allowed the antibodies to cross-react with the acrosomal enzymes in capacitating acrosome-intact spermatozoa without permeabilization, as revealed by the appearance of several distinct fluorescent patterns, including an initial immunopositive lining over the acrosome cap to an intense immunopositive reaction throughout the acrosome. These early immunopositive patterns were followed by the appearance of intense fluorescent spots (droplets) that seem to establish contact with the PM in a time-dependent manner. Inclusion of calmodulin, a 17-kDa Ca(2+)-binding protein which promotes capacitation, in the incubation medium did not alter the overall rate of capacitation; however, its presence accelerated the initial stages of membrane priming. The potential similarities between sperm capacitation and early events of Ca(2+)-triggered membrane fusion among eukaryotes and among various stations of the secretory and endocytotic pathways are discussed. PMID- 12649733 TI - Developmental changes in the ploidy of mouse implanting trophoblast cells in vitro. AB - Shortly after the onset of implantation, polar mouse trophoblast cells proliferate and give rise to the ectoplacental cone, constituted by two distinct cell populations: undifferentiated, diploid cells and giant cells. Giant cells characteristically exhibit exaggerated dimensions and polyploid nuclei. In this study, we employ ectoplacental cones as a dynamic source of trophoblast giant cells to analyze cell proliferation, cell death, and ploidy under in vitro conditions. Our results show that DNA synthesis and the increase in the cell number are relevant only during the first 24 h of culture. Subsequently, DNA synthesis still occurs, mainly in the giant cell compartment, while the number of cells gradually decreases. Cell death by injury and apoptosis was also observed in the non-giant cell compartment of the ectoplacental cone. These findings suggest that the first 24 h of culture are crucial to the mitotic activity of the ectoplacental cone cells that gradually ceases, favoring the endoreduplication process. The DNA synthesis index during the subsequent experimental intervals emphasizes accumulation of DNA for the polyploidization. There was clear correlation between DNA content and nuclear dimension. The ploidy values for the trophoblast giant cells varied from 2C up to 368C in the giant cells, but were not as expressive as those known from in vivo conditions, probably due to the absence of regulatory factors specific to the embryonic-maternal interface. In situ hybridization and histochemistry for the nucleolus-organizing region showed that trophoblast nuclei have only two marker signals, indicative of a typical polytenic process. This present study elucidates important aspects of trophoblast behavior and provides new information on trophoblast physiology in vivo and in vitro. PMID- 12649734 TI - Mosaic analysis of small intestinal development using the spf(ash)-heterozygous female mouse. AB - Mosaic analysis using the spf(ash)-heterozygous female mouse was performed to clarify the cell lineage and cell behavior during small intestinal development with special attention given to the villus and crypt formation. The spf(ash) mutation, located on the X-chromosome, causes ornithine transcarbamylase (OTC) deficiency, which leads to mosaic expression of this enzyme in the small intestine of the heterozygous female mouse. In the small intestine in heterozygous fetuses, very small patches, which were aggregates of OTC-positive cells or negative cells, with no definite orientation to the villus structures were observed. In the neonatal small intestine, the intervillus region (the presumptive crypts) was polyclonal, and the majority of crypts were comprised exclusively cells of either genotype in 2-week-old small intestine. These results suggest that extensive migration and cell mixing of small intestinal epithelial cells, which have no definite correlation with the villus formation, occur in fetal stages of development, and that the crypt morphogenesis commences after birth independently of the monoclonality of the epithelial cells. Our data with the mosaic mice also reconfirmed the monoclonality of the adult small intestinal crypts demonstrated in mouse aggregation chimeras. PMID- 12649735 TI - In bovine binucleate trophoblast giant cells, pregnancy-associated glycoproteins and placental prolactin-related protein-I are conjugated to asparagine-linked N acetylgalactosaminyl glycans. AB - Bovine binucleate trophoblast giant cells (BNCs) produce large amounts of PAS positive cytoplasmic granules. After fusion of BNCs with uterine epithelial cells, the contents of these granules are released into the maternal stroma which underlies the uterine epithelium. Histochemically, the granules can be labeled with N-acetylgalactosamine-specific lectins ( Dolichos biflorus, Vicia villosa, and Wisteria floribunda agglutinins) and with Phaseolus vulgaris leucoagglutinin. In this study, we used lectin western blot analysis of proteins from fetal cotyledons to characterize the lectin binding glycoproteins. Lectin western blots showed several bands. A main band of approximately 65 kDa was identified as pregnancy-associated glycoproteins (PAGs) and a double band at 34-35 kDa as prolactin-related protein-I (PRP-I) by their crossreactivity with specific antisera. Enzymatic cleavage of N-linked glycans with peptide- N-glycanase F abolished the lectin binding to PRP and PAGs in western blots, revealing that the lectins bound to asparagine-linked glycans. The high specificity of the lectins was used for the enrichment of PRP-I and PAGs from placental cotyledons with Vicia villosa lectin affinity chromatography. The occurrence of the relatively uncommon asparagine-linked N-acetylgalactosaminyl glycans on secretory proteins of the BNCs suggests a functional role of this specific glycosylation pattern. PMID- 12649736 TI - Developmental aspects of galectin-3 expression in the lens. AB - In order to investigate the temporal and spatial expression pattern of the lectin galectin-3 during lens development we performed immunohistochemical studies using monoclonal and polyclonal antibodies against galectin-3 on paraffin sections of human, mouse and rat eyes. Galectin-3 has been shown to be involved in various biological functions related to cell adhesion, proliferation, apoptosis and differentiation in other tissues. In the human lens, galectin-3 shows a selective expression pattern during lens development. It is present in all cells of the early lens vesicle and at later stages it is strongly expressed during the elongation phase in differentiating primary lens fibres. From about 7 weeks onwards the anterior lens epithelium fails to express galectin-3. Adult lenses, however, exhibit immunoreactivity in the anterior epithelial cells and in the early differentiating secondary fibres of the lens' outer cortex prior to the onset of degradation of the nuclei. In contrast to the observed expression pattern in prenatal human lenses, mouse and rat lenses exhibited immunoreactivity for galectin-3 during postnatal and adult stages only. At these stages, the expression pattern closely resembles that seen in the corresponding human lenses. The spatiotemporal pattern of galectin-3 distribution during lens development favours a role of this lectin in adhesion processes and in the regulation of programmed organelle elimination during lens cell differentiation. PMID- 12649737 TI - VEGF expression is developmentally regulated during human brain angiogenesis. AB - Vascular endothelial growth factor (VEGF) is a potent angiogenic factor working as an endothelial cell-specific mitogen and exerting a trophic effect on neurons and glial cells, both these activities being essential during central nervous system vascularisation, development and repair. The vascularisation of human telencephalon takes place by means of an angiogenic mechanism, which starts at the beginning of corticogenesis and actively proceeds up to the last neuronal migration, when the basic scheme of the vascular network has been drawn. Our study focused on VEGF during this critical developmental period with the aim of identifying the cells that express VEGF and of correlating the events of angiogenesis with the main events of cerebral cortex formation. The results show that in fetal human brain VEGF protein is located on multiple cell types, cells proper to the nervous tissue, neuroepithelial cells, neuroblasts and radial glia cells, and non-neuronal cells, endothelial and periendothelial cells. In these cells VEGF expression appears developmentally regulated and is correlated with angiogenesis, which in turn responds to the high metabolic demands of the differentiating neocortex. PMID- 12649738 TI - Expression and distribution of transcripts for sonic hedgehog in the early phase of fracture repair. AB - Localization and expression of mRNAs for sonic hedgehog (Shh) at a fracture site in the early phase postfracture were investigated by in situ hybridization and reverse transcription and polymerase chain reaction (RT-PCR). A closed fracture was made in the midshaft of the right tibia of 5-week-old ICR mice, and fractured sites were harvested prefracture (day 0) and on days 2 and 12. In situ hybridization revealed that transcripts for Shh were not detected on day 0, but they were detected in proliferating callus-forming cells in the periosteum and the surrounding tissue, and in the medullary cavity prior to apparent new cartilage and bone formation. Gli 1 (a signaling mediator for Shh) and bone morphogenetic protein-4 transcripts were colocalized with those for Shh transcripts on day 2. The RT-PCR showed that Shh mRNA was detected in the PCR product from day 2, but not from days 0 and 12. These findings are the first description about the activation of Shh gene in the early postfracture reaction. PMID- 12649739 TI - Coordinated regulation and colocalization of alphav integrin and its activating enzyme proprotein convertase PC5 in vivo. AB - Integrin alphav is involved in intracellular-extracellular signaling important for cytoskeleton alterations and control of cell movement. In vitro experiments indicate that the integrin alphav-subunit undergoes post-translational endoproteolytic cleavage. This type of activation requires the presence of suitable kexin/subtilisin-like proprotein convertases. In vitro experiments have demonstrated that, among several proprotein convertases, PC5A, and to a threefold lesser extent furin, can activate alphav integrin. The biological significance of these in vitro data would be further supported by a coexpression and coordinated regulation of the gene expression of alphav integrin and its activating enzyme PC5 in vivo. In the present study we investigated the regulation of alphav integrin and PC5 following balloon injury in vivo. Comparative immunocytochemistry revealed a coordinated regulation of alphav integrin and PC5 during vascular remodeling in rodents. Integrin alphav was found to be upregulated in PCNA-positive, proliferating vascular smooth muscle cells. Northern blots revealed no significant regulation of furin mRNA, whereas PC5A mRNA increased during vascular remodeling, suggesting that PC5 is the major convertase during neointima formation in vivo. Incubation of vascular smooth muscle cells with the Golgi-disturbing agent brefeldin A inhibited alphav integrin maturation, indicating that endoproteolytic cleavage occurs in the trans Golgi network, were PC5 is localized. Thus, the present study further supports the concept that activation of alphav integrin occurs in the trans-Golgi network in vascular smooth muscle cells and involves PC5. PMID- 12649740 TI - The WFS1 gene, responsible for low frequency sensorineural hearing loss and Wolfram syndrome, is expressed in a variety of inner ear cells. AB - Heterozygous mutations in the WFS1 gene are responsible for autosomal dominant low frequency hearing loss at the DFNA6/14 locus, while homozygous or compound heterozygous mutations underlie Wolfram syndrome. In this study we examine expression of wolframin, the WFS1-gene product, in mouse inner ear at different developmental stages using immunohistochemistry and in situ hybridization. Both techniques showed compatible results and indicated a clear expression in different cell types of the inner ear. Although there were observable developmental differences, no differences in staining pattern or gradients of expression were observed between the basal and apical parts of the cochlea. Double immunostaining with an endoplasmic reticulum marker confirmed that wolframin localizes to this organelle. A remarkable similarity was observed between cells expressing wolframin and the presence of canalicular reticulum, a specialized form of endoplasmic reticulum. The canalicular reticulum is believed to be involved in the transcellular movements of ions, an important process in the physiology of the inner ear. Although there is nothing currently known about the function of wolframin, our results suggest that it may play a role in inner ear ion homeostasis as maintained by the canalicular reticulum. PMID- 12649741 TI - Intracellular mislocalization of mutant podocin and correction by chemical chaperones. AB - The NPHS2 gene encoding the podocin protein was causally linked to the autosomal recessive type of steroid-resistant nephrotic syndrome. In this study, we investigated the consequence of the R138Q mutation of podocin, one of the most common missense mutations in the NPHS2 gene, by examining the expression of the wild-type and R138Q mutant podocins in mammalian cells. Either myc- or FLAG tagged wild-type podocin was strongly stained in plasma membrane, particularly in the fine processes wherein the protein was colocalized with actin stress fibers. On the other hand, the R138Q mutant podocin was completely retained intracellularly and colocalized with the endoplasmic reticulum (ER) marker, calnexin. These results suggest that the R138Q mutation affected podocin protein folding, thereby interfering with the mutant protein's departure from the ER. To determine if the ER retention of R138Q mutant is correctable, cells were incubated with the chemical chaperones glycerol, trimethylamine-N-oxide, and DMSO. Using these two methods, namely, cell surface labeling with sulfo-NHS-S-S biotin and Alexa 488-streptavidin, and immunostaining to detect the podocin protein close to the plasma membrane, we confirmed that these chemical chaperone treatments elicit a cellular redistribution of R138Q podocin. Our results reveal defective cellular processing of the mutant podocin, and provide evidence for pharmacological correction of the processing defect. PMID- 12649742 TI - IL-12: a promising adjuvant for cancer vaccination. AB - The clinical development of interleukin 12 (IL-12) as a single agent for systemic cancer therapy has been hindered by its significant toxicity and disappointing anti-tumor effects. The lack of efficacy was accompanied by, and probably related to, the declining biological effects of IL-12 in the course of repeated administrations at doses approaching the maximum tolerated dose (MTD). Nevertheless, IL-12 remains a very promising immunotherapeutic agent because recent cancer vaccination studies in animal models and humans have demonstrated its powerful adjuvant properties. Therefore, IL-12 may re-enter the arena of cancer therapy. Here, we review the immune modulating characteristics of IL-12 considered responsible for the adjuvant effects, as well as the results of animal and human cancer vaccination studies with IL-12 applied as an adjuvant. In addition, we discuss how studies with systemic IL-12 in cancer patients, and several other lines of evidence, indicate that IL-12 may exert optimal adjuvant effects only at low dose levels. Therefore, the MTD may not constitute the maximum effective dose of IL-12 for adjuvant application. PMID- 12649743 TI - Induction of cellular immunity by anti-idiotypic antibodies mimicking GD2 ganglioside. AB - Gangliosides are potentially useful targets for tumor destruction by antibodies. However, the role of gangliosides in T cell-mediated immunity to tumors is unclear. We produced three murine monoclonal anti-idiotypic antibodies (Ab2) against a monoclonal antibody (Ab1) that binds strongly to melanoma-associated GD2 ganglioside and weakly to GD3 ganglioside. All three Ab2 induced anti-anti idiotypic antibodies (Ab3) with Ab1-like binding specificity to tumor cells and antigen in rabbits. The Ab3 specifically bound to GD2(+) tumor cells and isolated GD2, and shared idiotopes with the Ab1. Two of the three Ab2 induced GD2-specific delayed-type hypersensitivity responses in BALB/c and C57BL/6 mice, but not in C57BL/6/CD4(-/-) mice. Peripheral blood mononuclear cells (PBMC) from a melanoma patient proliferated specifically in response to in vitro stimulation with Ab2. Proliferation was accompanied by Th1-type cytokine production. Our studies demonstrate the induction of ganglioside-specific T cell-dependent immunity by Ab2 in mice. These T cells showed specific reactivity to ganglioside expressed by tumor cells. PMID- 12649744 TI - A phase I study of autologous dendritic cell-based immunotherapy for patients with unresectable primary liver cancer. AB - BACKGROUND/AIMS: To evaluate the safety and feasibility of immunotherapy based on autologous dendritic cells (DC) for patients with unresectable primary liver cancer (PLC). METHODS: A total of ten patients were enrolled and immunized with DCs. Autologous DCs were generated ex vivo in the presence of granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). Cells were then pulsed with tumor lysate (TL), tumor necrosis factor-alpha (TNF-alpha) and keyhole limpet hemocyanin (KLH). Non-adherent cells were collected on day 9 and cells were administered into the inguinal lymph node. Each patient received 1 10 x 10(6) cells four times at weekly intervals. RESULTS: Immunization was well tolerated in all patients without significant toxicity. DC vaccination induced delayed-type hypersensitivity (DTH) against KLH in seven out of ten patients. In one patient, one of the two liver tumors (tumor in segment 7, 13 mm in diameter) decreased in size to 7 mm and showed necrotic change on computed tomography examination after eight immunizations. In two patients, serum levels of tumor markers decreased after vaccination. CONCLUSION: The present clinical trial suggested that immunization by TL-pulsed DCs is feasible in patients with unresectable PLC without any toxicity. Further improvement in the clinical results of immunotherapy might be expected by modifying the therapeutic protocol. PMID- 12649745 TI - Melphalan-induced up-regulation of B7-1 surface expression on normal splenic B cells. AB - We have previously shown that exposure of MOPC-315 or P815 tumor cells to the widely used anticancer drug melphalan ( L-PAM, L-phenylalanine mustard) leads to rapid up-regulation of B7-1 surface expression. Since B7-1-expressing tumor cells depend on B7-expressing host antigen presenting cells (APC) for the generation of CD8(+) T-cell-mediated antitumor immunity, and since L-PAM promotes the acquisition of tumor-eradicating immunity by CD8(+) T-cells from MOPC-315 tumor bearers, the current studies were undertaken to determine if L-PAM also up regulates B7-1 expression on host APC. Here we show that exposure of normal spleen cells to L-PAM leads within 24 h to up-regulated B7-1 expression on B220(+) cells (B cells). Studies into the mechanism through which L-PAM leads to up-regulated B7-1 expression revealed that within 2 h after exposure of normal spleen cells to L-PAM, accumulation of B7-1 mRNA is evident and this accumulation requires de novo RNA synthesis, indicating that the regulation is at the transcriptional level. The L-PAM-induced accumulation of B7-1 mRNA was prevented with the antioxidant N-acetyl- L-cysteine (NAC), indicating that reactive oxygen species are important for the transcriptional regulation. Although AP-1 and NF kappa B are considered redox-sensitive transcription factors, L-PAM led only to activation of NF-kappa B that bound specifically to a probe containing the corresponding binding site in the B7-1 gene. Moreover, selective inhibition of NF kappa B activation prevented the L-PAM-induced B7-1 mRNA accumulation, indicating that NF-kappa B activation is essential for L-PAM-induced B7-1 gene expression in normal spleen cells. Finally, in vivo administration of an immunopotentiating dose of L-PAM to normal mice was found to up-regulate B7-1 mRNA expression in their spleens. Thus, the ability of L-PAM to up-regulate B7-1 expression not only on tumor cells but also on host cells may contribute to the potentiating activity of L-PAM for the acquisition of CD8(+) T-cell-mediated tumor-eradicating immunity in tumor bearers. PMID- 12649746 TI - Analysis of in vitro immunization: generation of cytotoxic T-lymphocytes against allogeneic melanoma cells with tumor lysate-loaded or tumor RNA-transfected antigen-presenting cells. AB - In this study we compared several protocols for in vitro induction of cytotoxic T lymphocytes (CTL) from naive HLA-A*0201(+) peripheral blood mononuclear cells (PBMC) against allogeneic melanoma cells. As immunization material we compared: (1) the lysate of apoptotic or live melanoma tumor cells [MSM-M14 (M14) and MSM M3 (M3)]; and (2) total RNA extracted from the same melanoma cell lines, either unconjugated or conjugated with a charged carrier (DMRIE-C). Overall killing activity was very similar in CTL induced by tumor lysate or RNA. CTL induced by both methods preferentially killed an HLA class I-matched M14 melanoma cell line rather than HLA class I-unmatched M3 cells. Cytotoxicity could be partially blocked by anti-HLA class I antibodies. There were no significant differences in cytotoxicity and in other clonal characteristics in CTL lines induced by a lysate of apoptotic bodies as compared to lines induced by lysate of viable cells. However, CTL induced by DMRIE-C-bound total RNA demonstrated superior cytotoxicity when compared with CTL induced by unconjugated total RNA. Polyclonal CTL induced by tumor lysate contained a substantial percentage of tyrosinase(368 376 370N) tetramer-positive cells and demonstrated specific killing activity against tyrosinase(368-376 370N) peptide-labeled T2 cells, comparable to cytotoxicity of the CTL developed against this peptide alone. In contrast, there were no detectable tyrosinase(368-376 370N)-tetramer positive cells and no specific anti-tyrosinase peptide(368-376 370N) response in polyclonal CTL induced by immunization with tumor RNA. These data demonstrate that both total tumor RNA and tumor lysate are effective for inducing of cytotoxic anti-melanoma CTL, but tyrosinase(368-376 370N) specific cells were detected only in lysate-induced CTL cultures. This suggests that nature of the antigens present in tumor lysate might be different from those in tumor RNA. PMID- 12649747 TI - Comparison of the efficacy of local treatment of equine sarcoids with IL-2 or cisplatin/IL-2. AB - Local interleukin-2 (IL-2) is effective in a number of experimental animal models and in veterinary and human cancer patients without discomforting side effects. The primary goal of this study was to compare the therapeutic effects and side effects of the local intratumoral administration of five or ten low doses of IL-2 with those of a combination of cisplatin and a single high dose of IL-2 in the treatment of equine sarcoids. The therapeutic effect (complete and partial regression) of local cisplatin together with a single high dose of IL-2 was significantly better than the combined effect of low doses of local IL-2 administered daily over 5 or 10 days (80% and 43%, respectively; P=0.02). Cisplatin/IL-2 and low doses of IL-2 induced 53% and 14% complete regressions, respectively ( P=0.02). Histological changes after cisplatin/IL-2 treatment were far more pronounced than after IL-2 only treatment and in several cases showed an enormous eosinophilic infiltrate. PMID- 12649748 TI - Exposure to paclitaxel or vinblastine down-regulates CD11a and CD54 expression by P815 mastocytoma cells and renders the tumor cells resistant to killing by nonspecific cytotoxic T lymphocytes induced with anti-CD3 antibody. AB - Paclitaxel and vinblastine are drugs with anti-microtubule activity that are commonly used in the treatment of numerous types of cancer. In this study, we investigated the effect of prior exposure to submaximal cytotoxic concentrations (EC(25) and EC(50)) of paclitaxel or vinblastine on the subsequent susceptibility of surviving P815 murine mastocytoma cells to cytolysis by major histocompatibility complex (MHC)-unrestricted mouse cytotoxic T lymphocytes that had been induced with anti-CD3 antibody. P815 cells that had survived culture for 24 h in the presence of paclitaxel (5 or 50 micro g/ml) or vinblastine (1.5 or 15 micro g/ml) were rendered resistant to anti-CD3-activated killer-T (AK-T) cell mediated cytolysis in a standard (51)Cr-release assay. Resistance to killing was associated with a reduced ability of AK-T cells to form conjugates with drug treated P815 target cells, suggesting a possible effect on adhesion molecules. Flow cytometric analysis of paclitaxel- or vinblastine-treated P815 cells revealed reduced cell-surface expression of the adhesion molecules LFA-1 (CD11a /CD18) and ICAM-1 (CD54). Similar results were obtained following paclitaxel or vinblastine treatment of Yac-1 lymphoma cells. RT-PCR analysis revealed reduced levels of mRNAs coding for CD11a and CD54 in paclitaxel- or vinblastine pretreated P815 cells. Collectively, these data lead us to conclude that paclitaxel and vinblastine render P815 mastocytoma cells resistant to T cell mediated cytotoxicity by interfering with CD11a and CD54 expression by the tumor cells. A similar effect by these drugs on tumor cells and/or leukocytes in cancer patients might compromise tumor-specific cell-mediated immune responses. PMID- 12649749 TI - Cryopreservation of mature monocyte-derived human dendritic cells for vaccination: influence on phenotype and functional properties. AB - In the past decade there has been increasing evidence that tumor antigen-loaded dendritic cells (DC) are able to elicit anti-tumor T-cell responses. Initial clinical data for different tumor entities are encouraging, with objective tumor regressions being observed in some patients. Since GMP production of DC for clinical vaccination protocols is a time- and cost-intensive procedure, cryopreservation of DC in aliquots ready for clinical use would significantly facilitate DC-based vaccination in the clinic. We asked whether freezing and thawing alters the phenotype or functional properties of DC. DC from healthy volunteers and from patients with chronic myeloid leukemia (CML) were analyzed after freezing and thawing for their viability, morphology, immunophenotype (FACS profile), T-cell stimulatory capacity (mixed lymphocyte reaction) and mobility (time-lapse cinemicroscopy). Our results demonstrate that cryopreservation does not cause significant changes in the phenotype or function of DC, neither in DC from healthy volunteers nor in those from CML patients. Our data indicate that cryopreserved aliquots of DC are suitable for clinical application in DC-based immunotherapy protocols. PMID- 12649750 TI - Thigh cellulitis caused by toothpick ingestion. PMID- 12649751 TI - Sustainable chemistry: starting points and prospects. AB - We review here the concept of sustainable chemistry (SC), which is still in its early development. One important element of SC is commonly defined as chemical research aiming at the optimization of chemical processes and products with respect to energy and material consumption, inherent safety, toxicity, environmental degradability, and so on. An increasing number of assessment systems containing quantitative indicators for these aspects are currently being developed. In addition, however, SC should also address the societal aspect of sustainability. With respect to scientific research, the societal aspect is defined here by two requirements: (1) the assumptions, objectives and implications of chemical research and its technical application should be made more transparent to various societal actors; (2) uncertainty and ignorance should be treated more explicitly in the course of scientific research. Meeting these requirements is necessary in order to lift the division between the allegedly disinterested and non-normative scientific research and the value-laden sphere of societal needs, preferences and decision-making situations. This, in turn, is understood here as a contribution to a more sustainable scientific practice. We illustrate the two elements of SC-optimization of products and processes as well as including the societal aspect-with the examples of environmental chemistry, green chemistry and the environmental assessment of chemical products. While considerable progress has been made in these fields, the societal aspect of SC remains to be recognized more fully in all branches of chemical research. One prerequisite for this is the inclusion of SC into chemical education from the very beginning. PMID- 12649752 TI - Molecular biodiversity. Case study: Porifera (sponges). AB - Biological diversity--or biodiversity--is the term given to the variety of life on Earth and the natural patterns it forms. The biodiversity we see today is the fruit of billions of years of evolution, shaped by natural processes and, increasingly, by the influence of humans. It forms the web of life of which we are an integral part and upon which we so fully depend. The research on molecular biodiversity tries to lay the scientific foundation of a rational conservation policy that has its roots in various disciplines including systematics/taxonomy (species richness), present day ecology (diversity of ecological systems), and functional genetics (genetic diversity). The results of ongoing genome analyses (genome projects and expressed sequence tag projects) and the achievements of molecular evolution may allow us not only to quantitate the diversity of the present biota but also to extrapolate to their diversification in the future. A link between biodiversity and genomics/molecular evolution will create a platform which we hope may facilitate a sustainable management of organismic life and ensure its exploitation for human benefit. In the present review we outline possible strategies, using the Porifera (sponges) as a prominent example. On the basis of solid taxonomy and ecological data, the high value of this phylum for human application becomes obvious, especially with regard to the field of chemical ecology and the desire to find novel potential drugs for clinical use. In addition, the benefit of trying to make sense of molecular biodiversity using sponges as an example can be seen in the fact that the study of these animals, which are "living fossils", gives us a good insight into the history of our planet, especially with respect to the evolution of Metazoa. PMID- 12649754 TI - Landmark memories are more robust when acquired at the nest site than en route: experiments in desert ants. AB - Foraging desert ants, Cataglyphis fortis, encounter different sequences of visual landmarks while navigating by path integration. This paper explores the question whether the storage of landmark information depends on the context in which the landmarks are learned during an ant's foraging journey. Two experimental set-ups were designed in which the ants experienced an artificial landmark panorama that was placed either around the nest entrance (nest marks) or along the vector route leading straight towards the feeder (route marks). The two training paradigms resulted in pronounced differences in the storage characteristics of the acquired landmark information: memory traces of nest marks were much more robust against extinction and/or suppression than those of route marks. In functional terms, this result is in accord with the observation that desert ants encounter new route marks during every foraging run but always pass the same landmarks when approaching the nest entrance. PMID- 12649753 TI - Hydrocarbon synthesis by enzymatically dissociated oenocytes of the abdominal integument of the German Cockroach, Blattella germanica. AB - In insects, hydrocarbons waterproof the cuticle, protect the insect from the external environment, and serve as semiochemicals or their metabolic precursors. In the German cockroach, Blattella germanica, hydrocarbons are synthesized by the abdominal integument, but the precise site of biosynthesis is not known. We developed a method for separation of oenocytes from other cells in the abdominal integument using enzymatic dissociation followed by Percoll gradient centrifugation. Radiolabeled propionate was then used to monitor de novo synthesis of hydrocarbons by dissociated cells. Oenocyte-enriched cell suspensions of abdominal sternites synthesized hydrocarbons, whereas suspensions enriched with epidermal cells did not. Our results show conclusively that hydrocarbons are produced by oenocytes not only in insects whose oenocytes are localized within the hemocoel, but also in those insects whose oenocytes are within the abdominal integument. Furthermore, these data support a hemolymph pathway for transport and delivery of hydrocarbons to both external and internal tissues, including the epicuticle, fat body, and ovaries. PMID- 12649755 TI - Extinction and re-evolution of similar adaptive types (ecomorphs) in Cenozoic North American ungulates and carnivores reflect van der Hammen's cycles. AB - Numerous patterns in periodicity (e.g., climate, extinction, and sedimentary cycles) and evolutionary change (e.g., chronofaunas and coordinated stasis) have been described based on aspects of the geologic record. Recently, convergent occurrences of faunal types or "repeating faunas" have received attention, but a highly specific, iterative pattern was first reported over 40 years ago. In the late 1950s, van der Hammen described climatic/floral cycles on the order of six million years based on a succession of A, B, and C pollen community types in South America. These A-B-C cycles are also seen in the replacement pattern of particular carnivore and ungulate adaptive types in Cenozoic North America as reported by Martin in the 1980s. For example, in the last 36 million years, there were four iterations of a sabertooth cat ecomorph independently evolving, dominating the niche through an A-B-C cycle, and then going extinct. Here we show further support for the existence of these cycles in the dominance turnover in hippo and dog ecomorphs in the North American Cenozoic. Shared patterns of extinction and re-evolution of adaptive types among plants and mammals across two continents suggest a global mechanism, which appears to be climatic change. Iterative climatic cycles of various scales may form a predictive framework for understanding fundamental patterns in the geologic record, such as radiations, extinction, rates of change, convergence, and sedimentary cycles. PMID- 12649756 TI - Microbial spheres: a novel cyanobacterial-diatom symbiosis. AB - Cyanobacteria, algae and bacteria are widespread inhabitants of North Sea microbial mats. Our studies of these populations showed uncommon modes of living and extraordinary structures, which have not been described before. The structures are spherical objects covering a community of cyanobacteria, diatoms and bacteria. The cultivation of these communities in the laboratory and intensive observations of their exceptional movement has led to some spectacular findings. The sphere formations go through different phases with variation in the dominance of different microorganisms. The role of the bacteria is the most important in the first phase, and can be increased by the addition of signal substances. Spheres surrounded by envelopes of unknown composition and permeability appear, with numerous bacteria and sporadic diatoms inside. Then the cyanobacteria penetrate the spheres and arrange themselves at the surface. The communities proliferate over some weeks and are finally released. Laboratory expositions of the microbial communities to different parameters pinpoint the limits of sphere formation. The metabolic products of the sphere communities are concentrated in the spheres and lead to a different kind of compound compared with the surrounding environment. In this way, the microbial communities strongly influence the structure of the sediments. Uncommon circular structures, which develop into spheres between 0.08 and 3 mm in size were found in subcultures of non-axenic filamentous cyanobacteria enrichments from North Sea microbial mats. These filamentous cyanobacteria ( Phormidium sp.) together with associated benthic diatoms of the genus Navicula and associated heterotrophic bacteria were held as reproducible synergistic cultures. Phormidium sp. filaments tightly intertwined with each other, formed the surface of the spheres, trapping diatoms inside. The formation of "spheres" was the result of radial and synchronous movements of the cyanobacteria. In old cultures, the direction of the cyanobacterial movement has turned in the opposite direction, away from the sphere. The integrity of large "spheres" was influenced by chemotactic phenomena and maintained by some type of trichome-trichome interaction. This suggests the presence of metabolic secondary products, which attract cyanobacteria and influence their movement in a form of chemotactic response. PMID- 12649757 TI - Intersexual dominance, masculinized genitals and prenatal steroids: comparative data from lemurid primates. AB - Masculinization of female genitalia and female intersexual dominance distinguish spotted hyenas (Crocuta crocuta) and Malagasy primates (Lemuriformes) from most other mammals. An unusual prenatal endocrine environment has been proposed to proximately underlie the development of these traits in hyenas. To examine whether female dominance and genital masculinization are similarly enhanced by the prenatal environment in lemurid primates, we measured androgen and estrogen excretion in pregnant wild redfronted lemurs (Eulemur fulvus rufus). Our results showed that estrogen levels during the second phase of gestation were much higher in females carrying a male fetus than in female-carrying mothers. This may indicate the onset of testicular activity in male fetuses, because androgens of fetal origin are aromatized to maternal estrogens. Levels of androgen excretion were similar in all mothers regardless of the fetus' sex, which may suggest that androgen-independent mechanisms also contribute to female masculinization. The much higher androgen/estrogen ratio in female-carrying mothers indicates that relative, rather than absolute, prenatal steroid concentrations may play a role in female masculinization. PMID- 12649758 TI - [Demand and supply in psychotherapy]. PMID- 12649759 TI - [The state of outpatient psychotherapy in Germany]. AB - This paper presents the results of a study of the state of adult outpatient psychotherapy in Germany after the PTG came into force. 1042 psychotherapists were questioned on certain issues. One result was that patients have to wait 4.6 months for psychotherapy and that every second patient asking for a diagnostic interview and possible treatment was refused. Of those who were given a diagnostic interview 35 % were not taken into treatment, although disturbances were diagnosed--such as tinnitus, pain, organic disturbances with psychic complications, suicidal tendencies, anorexia nervosa, addiction, psychosomatic illnesses, personality disorders, psychotic disorders--would normally demand psychotherapeutic treatment. Furthermore only 56 % of those patients who Locherbach et al. considered needing and wanting psychotherapeutic treatment were actually in a G IV psychotherapy. Apart from this the possibility of getting psychotherapy as well as the kind of psychotherapy proved to be dependent on the kind of medical insurance. Different payments by the insurance companies caused longer waiting times for patients and determined the choice of psychotherapy by the psychotherapists. PMID- 12649761 TI - [Changes in clinically relevant goals and therapy outcome: a study with inpatients undergoing cognitive behavioral therapy]. AB - According to the consistency theory clinically relevant goals, as explicated by Grosse Holtforth and Grawe, play a prominent role in the development, maintenance and therapy of psychological disorders. Thus effective psychotherapies should "normalize" the subjective importance of these goals. These changes should be positively associated with the success of psychotherapy. To test these assumptions, clinically relevant goals of 64 inpatients undergoing cognitive behavioral therapy were assessed by the Inventory of Approach and Avoidance Goals (German: Fragebogen zur Analyse motivationaler Schemata, FAMOS) before and after therapy. Results show effects of normalization, as expected, especially on scales associated with psychopathology in former studies. PMID- 12649760 TI - [Therapy goals, change of goals and goal attainment in the process of psychoanalytically oriented and behavior long-term therapy--a comparative study from the private practices of insurance-registered psychotherapists]. AB - In a prospective naturalistic design 31 patients with long-term behavior therapy (average 63 treatment hours) and 31 patients with psychoanalytically-oriented long-term therapy (average 185 treatment hours) were compared. All patients were examined by extern interviewers with the Structured Clinical Interview for Diagnosis (SCID) before they were included in the study. Only patients that showed DSM-III-R criteria of a depressive or anxiety disorder were included. At four times the patient goals in therapy and the actual state was examined: At the beginning of treatment, after 1, after 2,5 and after 3,5 years. At all times the patients could formulate new goals and release old ones. The symptoms were registered by SCL-90-R and the interpersonal problems were recorded by IIP-D. After 3,5 years a follow-up interview was conducted. Although all patients were comparable in their diagnoses the patients in behavior therapy and those in psychoanalytically-oriented therapy differed in a number of characteristics, for example by the way they gained access to therapy (doctors' subscription versus personal initiative for an appointment), the education, the consuming of psychotropic medication and the strain of symptoms. The patients did not differ remarkably in their goals in therapy. The symptoms as a goal did not have the highest priority in both groups, but the category "self-worth-problems" was found in both groups with a high priority. About one third of therapy goals were redefined by the patients in both groups within one year. After 2,5 years the number of goals in the category "interpersonal conflicts" increases remarkably in both treatment groups. For both groups we found the extent of reaching aimed goals in therapy (recorded by GAS) was significant over time. The results seem to prove that psychotherapy under naturalistic conditions aims more at improving the general level of functioning than at reducing the symptoms. PMID- 12649762 TI - [Psychotherapy within the prison system after the change of law]. AB - The amendment of the law limiting the professional secrecy of physicians and psychologists toward the prison authorities has triggered a discussion on the consequences involved for treatment practice in the prison system. To reduce the lack of empirically based arguments in this discussion, our study deals with the question of whether and to what extent psychotherapy in conjunction with prison health care offers also contributions to preventing recidivism. A quasi experimental control group study showed that especially for serious violent offenses delinquents of the Tegel detention center who underwent at least 20 hours of therapy at the psychotherapeutic consultation and treatment service had a markedly lower recidivism rate a mean of four years after release from prison than an untreated group. If changes in the penal code jeopardize a regular therapeutic setting with professional secrecy, they carry the risk of excluding prisoners primarily requiring psychotherapy who have thus far apparently also derived benefit especially from prevention-oriented therapy. PMID- 12649763 TI - [Relationship of sick leave before treatment to severity of symptoms and treatment outcome in in-patients with anxiety disorders]. AB - The aim of this study is to explore whether or not being already on sick leave at admission to a psychosomatic clinic indicates a higher level of severity of symptoms in patients with anxiety disorders, and whether or not this has an impact on therapy outcome. We examined 194 in-patients at 8 psychosomatic clinics upon admission and discharge by interview and psychometric testing. Being on sick leave before admission proved to be an indicator for higher global symptom distress as well as a higher severity of anxiety symptoms. Treatment duration was longer for the sick leave group than for the patients that had not been on sick leave, but each group experienced the same degree of change in pre-and-post treatment symptoms. We conclude that sick leave before admission does give information about illness severity and need of treatment in patients with anxiety disorders. PMID- 12649764 TI - [Traumatic and psychosocial distress in patients with acute tumors]. AB - For patients, being diagnosed with cancer means being confronted with a life threatening illness and can thus be considered a traumatic experience. This situation is compounded by the fact that having cancer is usually not an isolated event but is generally a long-term stressor. The aim of the present study was a) the assessment of psychotraumatic and comorbid distress in tumor patients both in the acute stage (T1) and over time (T2), b) the identification of possible protective factors, and c) the identification of risk factors for the development of a posttraumatic stress reaction (PTSR) following the diagnosis of a tumor. The sample consisted of 46 patients who had to undergo surgical treatment for a newly developed tumor for either diagnostic or therapeutic purposes. The following psychosocial factors were examined: posttraumatic stress, dissociative reactions, anxiety, depression, cognitive-emotional distress, social support and processing the illness. More than 25 % of the patients demonstrated signs of a clinically relevant PTSR at the acute stage (T1) and almost 40 % did so four months later (T2). Significantly more women showed a PTSR at both points of measurement, but particularly at follow-up. No significant relationship could be found between the type of the tumor at T1 and the emergence of a PTSR at T2. Patients with PTSR at T2 showed higher scores for anxiety and depression. The most important predictors for the development of clinically relevant posttraumatic symptoms at the four month follow-up examination were female sex, PTSR at T1 as well as elevated levels of depression. The results indicate the clinical relevance of psychotraumatic screening examinations in patients with acute cancer illnesses both for diagnostic as well as therapeutic investigations. PMID- 12649767 TI - Melperone is an inhibitor of the CYP2D6 catalyzed O-demethylation of venlafaxine. AB - INTRODUCTION: Melperone, a butyrophenone neuroleptic, is frequently used for its sleep-inducing properties. Despite its common use for more than 30 years, it is not yet characterized regarding its effects on cytochrome P450 s (CYPs). In an open pilot study, effects of melperone on the steady-state blood levels of venlafaxine, a recently introduced serotonin- and noradrenaline reuptake inhibiting antidepressant, were assessed. METHODS: The dose-corrected serum concentrations of venlafaxine and O-desmethylvenlafaxine were analyzed retrospectively in a therapeutic drug-monitoring (TDM) database comprising 94 patients. In addition, three patients received venlafaxine and melperone concomitantly and the serum concentrations of venlafaxine and O desmethylvenlafaxine were analyzed before, during, and after melperone co medication. The effect of melperone on CYP2D6 was further assessed in seven patients by means of the dextromethorphan O-demethylation, which serves as a CYP2D6 probe reaction. RESULTS: Patients treated concomitantly with venlafaxine and melperone had significantly higher (mean +/- SD) venlafaxine (3.27 +/- 2.9 vs. 0.97 +/- 0.99 ng/ml per mg/d; p < 0.05) and lower O-desmethylvenlafaxine serum concentrations (0.69 +/- 0.35 vs. 1.51 +/- 0.9 ng/ml per mg/d; p < 0.01) compared to patients without melperone comedication. In the three patients, venlafaxine serum concentrations increased, on average by 52 % during melperone co-medication, whereas O-desmethylvenlafaxine was decreased, on average by 29 %. Administration of melperone over three days elevated the ratio of dextromethorphan to dextrorphan from 0.044 +/- 0.04 to 0.09 +/- 0.083 (p < 0.05). DISCUSSION: This study pointed to an inhibitory effect of melperone on the O demethylation of venlafaxine. Because the O-demethylation of venlafaxine is almost exclusively catalyzed by CYP2D6 it is concluded that melperone is an inhibitor of CYP2D6. The hypothesis was further corroborated by the inhibitory effect of melperone on the dextromethorphan O-demethylation. PMID- 12649768 TI - Anxiety induced by repetitive transcranial magnetic stimulation is suppressed by chronic treatment of paroxetine in rats. AB - Repetitive transcranial magnetic stimulation (rTMS) has been shown to have a therapeutic effect on affective disorder and anxiety disorders. However, some reports have linked rTMS to a significant increase in anxiety in normal volunteers. This study investigates the effect of rTMS on anxiety and the use of acute and chronic paroxetine treatment on this animal model of anxiety. In normal rats, rTMS for 10 days induced anxiety, as shown by elevated plus maze, black and white box, and conditioned fear tests. This anxiety was suppressed by chronic, but not acute, paroxetine. These results suggest that rats receiving chronic rTMS treatment can be used as a model of anxiety and that the anxiety induced by rTMS might involve the serotonergic system. PMID- 12649769 TI - Clinical outcome after trimipramine in patients with delusional depression - a pilot study. AB - The treatment of delusional depression is a major challenge in psychopharmacology. Hypothalamic-pituitary-adrenocortical (HPA) overdrive may contribute, via increased dopaminergic activity, to the pathophysiology of the disorder. Trimipramine appears to be an interesting potential candidate, since it is an atypical antidepressant that is known to inhibit HPA activity. In a four week open trial we investigated its effects in 15 inpatients with delusional depression. The dosage was increased within 7 days up to 300 - 400 mg/d and was then maintained for three weeks. Psychometric assessments and safety monitoring were conducted weekly. Assessment of the HPA activity was achieved by a combined dexamethasone suppression/corticotropin-releasing hormone stimulation (Dex/CRH) test before and after four weeks of treatment. Therapeutic response was defined as a decrease in the HAMD-score of at least 50 %. Eight out of 13 completers were rated as responders. Therapeutic response was associated with L, D-trimipramine concentrations higher than 160 ng/ml. Intent-to-treat analysis showed significant improvement in psychometric variables. Despite the high dosage, the substance was generally well tolerated, with the exception of one patient who suffered from a hypotensive reaction. Mean +/- SD concentration of L-trimipramine and D trimipramine were 138 +/- 61 ng/ml and 119 +/- 50 ng/ml at a final dose of 346 +/ 50 mg/d. The ACTH and cortisol area under the curve in the Dex/CRH tests decreased significantly, reflecting a decrease of activity in the HPA system. We suggest that the clinical use of high-dose trimipramine in delusional depression seems to be a promising treatment strategy. PMID- 12649770 TI - Are the second-generation antipsychotics cost-effective? A critical review on the background of different health systems. AB - BACKGROUND: Despite clinical advantages over conventional compounds, second generation antipsychotics are prescribed less frequently in some European countries than in the United States because of their higher acquisition price and the current cost-containment strategies of many European health systems. This has been criticized on the grounds that the higher acquisition costs of the new antipsychotics might be more than outweighed by savings in other fields, e. g., through a reduction in rehospitalizations or indirect costs. METHOD: In order to create an empirical basis for this discussion, a review of the results of pharmacoeconomic studies (mostly cost-effectiveness studies) comparing second generation with conventional antipsychotics was undertaken. RESULTS: Of the 35 studies identified, most report at least cost-neutrality of the new antipsychotics (in many cases clozapine) that is due to reductions in hospitalization costs. These results cannot be generalized, however, because of methodological shortcomings such as small patient samples and study designs with low validity, and especially because of a lack of studies performed outside the U.S. It is shown that results from studies in the U.S. cannot be generalized to other health systems in Europe or in developing countries. Furthermore, only a few findings on newer second-generation antipsychotics other than clozapine are reported, and no study investigated indirect costs, which play a major role because of the early onset and chronicity of schizophrenia. CONCLUSION: Until now, there has been no sufficient evidence for the superior cost-effectiveness of atypical antipsychotics in European countries. Considering the importance of this topic for health politics, more cost-effectiveness studies in European countries are urgently needed. But even if economic superiority of the second-generation antipsychotics cannot be demonstrated in such studies, their use is nevertheless indicated with respect to patient's well-being. PMID- 12649771 TI - The high-affinity non-peptide CRH1 receptor antagonist R121919 attenuates stress induced alterations in plasma oxytocin, prolactin, and testosterone secretion in rats. AB - Evidence from basic and clinical research suggests that hyperactivity of central corticotropin-releasing hormone (CRH) circuits contributes to causality and course of affective disorders. Therefore, CRH receptor antagonists have attracted attention as potential therapeutics. We could previously show that the novel high affinity non-peptide CRH 1 receptor antagonist R121919 significantly inhibits stress-induced corticotropin release and displays anxiolytic effects in rats selectively bred for high anxiety-related behavior. These animals are characterized by their innate hyper-reactivity of the hypothalamic-pituitary adrenocortical system linked to an increased emotionality and therefore are suitable for the evaluation of CRH 1 receptor antagonists. Here we show that in addition to its effects on anxiety-related behavior and corticotropin secretion, R121919 attenuates the stress-induced release of corticosterone, prolactin, and oxytocin. Moreover, the decrease in plasma testosterone following exposure to stress is abolished by R121919. Our data indicate that antagonism of CRH 1 receptors may prevent stress-associated endocrine alterations. PMID- 12649772 TI - Hypertension induced by St. John's Wort - a case report. PMID- 12649773 TI - Minimal myocardial damage after tricyclic neuroleptic overdose - a case report. AB - A great number of acute, severe self-poisoning cases are due to tricyclic neuroleptic overdose. These drugs may induce several effects on the myocardium, mainly related to arrhythmias and electrical conduction and excitation abnormalities. These abnormalities are particularly evident after massive ingestion of these drugs for suicidal purposes. More recently, abnormal ventricular repolarization, in association with QRS widening, that mimics an acute ischemic event has been reported. However, these transient ECG abnormalities have never been associated with organic cardiac alterations. We report the case of a young woman who - according to ECG, enzyme release, and echocardiography - developed transient acute myocardial ischemia and minimal myocardial damage following self-poisoning with promazine hydrochloride. To our knowledge, this is the first case of documented minimal myocardial damage after tricyclic drugs overdose. PMID- 12649774 TI - Methadone maintenance treatment and St. John's Wort - a case report. AB - St. John's wort, a popular over-the-counter drug for treatment of depression, might reduce concentrations of drugs such as cyclosporin and indinavir and lead to drug resistance and treatment failure. No studies as yet have examined its influence on methadone plasma levels. The trough methadone plasma levels were measured in four patients (2 males, median age: 31 years; range 19 - 40 years) in methadone maintenance treatment just before the introduction of St. John's wort (900 mg/d) and after a median period of 31-day treatment (range 14 - 47). The study was proposed to addict patients about to start an antidepressant therapy. Introduction of St. John's wort resulted in a strong reduction of (R,S)-methadone concentration-to-dose ratios in the four median patients included, with a median decrease to 47 % of the original concentration (range: 19 % - 60 % of the original concentration). Two patients reported symptoms that suggested a withdrawal syndrome. Thus, prescription of St. John's wort might decrease methadone blood levels and induce withdrawal symptoms which, if not correctly identified and handled (by changing the antidepressant or by increasing the methadone dose), might cause unnecessary discomfort to the patient, lead to resumption of illicit drug uses, or be a risk factor for discontinuation of the methadone or antidepressant treatment. PMID- 12649775 TI - Increased apoptosis of neutrophils in a case of clozapine-induced agranulocytosis - a case report. AB - A 45-year-old female suffering from severe chronic schizophrenia of the paranoid type did not respond to typical antipsychotics. Five weeks after starting therapy with clozapine, she developed a clozapine-induced agranulocytosis (CA). Discontinuation of clozapine and treatment with granulocyte colony-stimulating factor (G-CSF) led to normalization of blood neutrophil counts within three weeks. This report suggests enhanced apoptosis of blood neutrophils during the acute phase of CA resulting from enhanced expression of the pro-apoptotic proteins Bax and Bik and from a decrease of the anti-apoptotic BCl-X(L) mRNA. The time course of decline and recovery of neutrophilic cells, as well as the release pattern of endogenous G-CSF, resembles those of chemotherapy-induced neutropenia. The kinetics of CD 34-positive cells mimics that of cytotoxic progenitor cell mobilization, e. g., after cytostatic drug administration. Our findings argue against the hypothesis that clozapine-mediated inhibition of G-CSF or granulocyte macrophage colony-stimulating factor (GM-CSF) release is involved in CA development. Because clozapine-induced cell death mainly affects the neutrophil lineage, the elucidation of the exact mechanism of CA may open new perspectives for the treatment of psychiatric and possibly hematological disorders. PMID- 12649776 TI - Fluoxetine-induced exacerbation of chorea in Huntington's disease? A case report. PMID- 12649777 TI - Effects of anticholinergic drug withdrawal on memory, regional cerebral blood flow and extrapyramidal side effects in schizophrenic patients by K. Mori et al. PMID- 12649779 TI - [Are fatal hemorrhagic complications during and after birth preventable?]. PMID- 12649780 TI - [Solitary and multiple chorangiomas--clinical consequences, expression of growth factors and differences in the growth rate]. AB - BACKGROUND: Chorangiomas are regarded as hamartous lesions of the placenta which may complicate a pregnancy if they grow large. The etiology of these lesion is still unclear. We suspected a link between the development of chorangiomas and an increase of the expression of the angiogenic growth factor bfgf and angiopoietin 1 within the tumour. MATERIAL AND METHODS: We examined 20 placentas without tumour, 19 placentas with solitary chorangiomas and 10 placentas with multiple chorangiomas (chorangiomatosis) with respect of the difference in clinical complications, regarding the expression of bfgf and angiopoietin-1 and with regard to the proliferation rate. RESULTS: The expression of the growth factors in solitary chorangiomas did not differ from that in the normal placental tissue. Both groups showed moderate expression of growth factors. In placentas with multiple chorangiomas all cases were associated with a strong expression of bfgf and angiopoietin-1. Proliferating cells and fibroblasts were seen more often in placentas with chorangiomatosis. There were clinical differences with regard to the maternal age. The mean age in case of normal placentas was 24 years, in placentas with solitary chorangiomas 32 years and for placentas with multiple chorangiomas 28 years. There were more complications such as HELLP-syndrome, diabetes, preterm birth and additional maturational arrest of the placenta in case of chorangiomas. CONCLUSION: There is a link between the development of multiple chorangiomas and an increase of the expression of bfgf and angiopoietin 1 in the placenta. In our study there is no correlation between multiple tumours and complications of pregnancy. PMID- 12649781 TI - [Supply of iodine during pregnancy--an inventory in Berlin, Germany]. AB - BACKGROUND: Iodised salt was introduced in Germany in the early 1980s. A nation wide study in 1996 showed that iodine levels among the population had improved since the introduction of the supplementation. The study did not separately investigate the iodine status of pregnant women. In our prospective study, we used three parameters to assess the iodine levels among pregnant women. PATIENTS AND METHODS: Between October 1999 and February 2000, we asked 109 German-speaking patients seeking prenatal care in our clinic to participate. Following informed consent, we measured goiter volume by ultrasound and collected venous blood (serum) and urine samples. We asked patients about any history of thyroid gland illnesses and about iodine supplementation which is generally given to pregnant patients in Germany. The blood and urine samples were stored at -18 degrees C until measurement. We used the iodine-creatinine-ratio to measure ioduria. Iodine was measured using the Cer-Arsenite-method (Dade-Behring). The thyroglobulin concentration in serum was measured using RIA. RESULTS: The mean iodine creatinine ratio was 181 +/- 109 microg/g, 20.4 % of the patients had a ratio between 50 and 100 microg/g which is defined as iodine deficiency I degrees (WHO). 8.7 % of the patients had thyreoglobulin levels above the cut-off value of 50 ng/ml. 12.6 % of the patients had a goiter (volume > 18 ml). 58 % of the patients were taking iodine supplements. These patients had significantly higher iodine-creatinine ratio levels (204 microg/g vs. 148 microg/g, P = 0.007) and lower serum thyroglobulin levels (38.4 vs. 34.1 pmol/l, P = 0.06) than non supplemented patients. CONCLUSIONS: The prevalence of goiter reflects an extended period of iodine deficiency. Using laboratory methods, up to 20.4 % of pregnant women were identified as having an iodine deficiency which indicates the need for a general iodine supplementation during pregnancy. PMID- 12649782 TI - [Scientific evidence on labor duration from systematic reviews]. AB - OBJECTIVE: We examined the evidence of systematic reviews in the Cochrane Library on interventions that influence the duration of labor. MATERIAL AND METHODS: Of the 208 Cochrane Pregnancy and Childbirth systematic reviews, 69 deal with labor and birth of which 20 address the effect of intrapartum interventions. We analyzed the reviews with respect to those interventions that affected the duration of birth. RESULTS: Six reviews, related to 5 interventions (amniotomy, epidural analgesia, continuous support, amnioinfusion, and maternal position), reported a significant effect on the duration of labor. Six found no significant effect on labor duration. The remaining eight reviews did not assess the effect of the intervention on labor duration. DISCUSSION: Interventions that have a significant influence on labor duration may interact with each other. Therefore, perinatal databases should ascertain not only if but also when an intervention has been performed. PMID- 12649783 TI - [Risk factors of retinopathy of prematurity in infants 32 to 36 weeks gestational age]. AB - INTRODUCTION: In our study we determined possible risk factors for retinopathy of prematurity (ROP) in infants 32 to 36 completed weeks of gestational age based on a regional German neonatal database. We examined especially whether or not oxygen therapy over more than 3 days is related to a higher risk of ROP. MATERIALS: We identified 7172 ophthalmologically examined infants, 32 to 36 completed weeks gestational age, born from 1990 to 1996. ROP was diagnosed in 195 (2.7 %). We examined the following variables as risk factors for ROP in infants receiving oxygen for less than 4 days: gestational age, sex, blood pH of 7.0 or less, body temperature of 36 degrees C or less, phototherapy, blood pO2 of 35 mm Hg or less, small-for-gestational age, sepsis, ventilation after birth, and blood transfusion. RESULTS: Sex, blood pH of 7.0 or less, blood pO2 of 35 mm Hg or less, sepsis, phototherapy, and small-for-gestational age were not associated with a significant risk of ROP. A gestational age of 32 weeks compared to a gestational age of 36 weeks was associated with an increased risk of ROP (odds ratio, 2.95; 95 % confidence interval, 2.18 to 4.01). Ventilation after birth (adjusted OR, 2.29; 95 % CI, 1.70 to 3.15) and blood transfusion (adjusted OR, 5.28; 95 % CI, 3.80 to 7.23) increased the risk of ROP regardless of gestational age. Oxygen therapy for more than 3 days was not associated with an increased risk of ROP (OR, 1.06; 95 % CI, 0.67 to 1.70). CONCLUSION: In neonates delivered between 32 and 36 weeks of gestation, the duration of oxygen supplementation should not necessitate an ophthalmological examination. A vigorous restriction of blood transfusions could reduce the incidence of ROP. PMID- 12649784 TI - [Prenatal diagnosis of hepatic hemangioendothelioma and peripartual management]. AB - Infantile hepatic hemangioendothelioma is a benign hepatic tumor that can be associated with life-threatening perinatal complications. A 38-year-old gravida 3 para 2 was referred to our hospital with preterm labor at 33.6 gestational weeks. Fetal abdominal circumference was estimated sonographically above the 97 th percentile. Because of pathologic CTG, the fetus was delivered by caesarean section (APGAR-scores 0/0/7, pH: 7.15). After cardiopulmonary reanimation and intensive care of the fetus, the prenatally suspected hemangioma of the liver was confirmed by computer tomography. The day after, liver segments V, VI, and VII were resected because of lung compression. During the operation, reanimation was necessary two more times. Two days later, ductus arteriosus Botalli was occluded. After stabilization, the neonate was treated with prednisone because of tumor residues. If a large hyperechogenic mass in the fetal liver is seen prenatally, diagnosis of fetal hemangioma should be considered. In order to prevent serious cardiovascular complications, continual clinical surveillance is indicated. PMID- 12649785 TI - [Empirical report on acute twin-twin transfusion syndrome with single intrauterine death]. AB - We report on a case of acute twin-twin transfusion syndrome in a twin pregnancy in the 26 th under primarily unclear conditions. On admission to the hospital, one fetus was not showing signs of life anymore, while the Doppler indices and CTG of the living fetus showed signs of acute distress. On the scan both fetuses showed adequate and symmetric growth as well as symmetric and normal amniotic fluid amounts, indicating a lack of typical signs for chronic twin-twin transfusion syndrome. The emergency cesarean section performed under the assumption of acute twin-twin transfusion syndrome, which unfortunately could not save the second twin, confirmed our suspected diagnosis. PMID- 12649786 TI - [Advanced training and responsibility]. PMID- 12649787 TI - [Quality management in and by public health services]. AB - Quality management is nowadays required by almost all organisations using public funds. Hence, quality management in public health services (PHS) first serves to make transparent the services offered, the performance, and the quality of services to comply with public demands for accountability. In addition, by following generally accepted concepts and methods of quality management, it could help to improve the performance of PHS. As a second function, quality management by PHS can be delineated. In this, PHS assumes the role of an independent quality manager of health services by reporting comparatively on regional services, their performance and the quality of care on a population basis. Considering recently introduced statutory incentives in Germany (such as disease management programmes and minimum volume thresholds for certain procedures) that could negatively affect social equity and access to health services, such a comparative quality and performance reporting by PHS is getting more and more important. PMID- 12649788 TI - [First evaluation of the surveillance systems of notifiable diseases under the infectious disease control law in Germany]. AB - INTRODUCTION: The implementation of the infectious disease control law (IfSG) in 1.1.2001 standardised the German surveillance system for notifiable diseases. For management and transmission of reports health departments use either the software programme Surv Net@RKI, which was developed by the Robert Koch-Institut (RKI), or one of five commercially offered disease-reporting software. After more than one year of its existence, we investigated the success of the implementation of the new surveillance system with the aim to identify possibilities for further improvement. METHODS: Based on 2001 data available to the RKI, we evaluated the criteria simplicity (standardisation of legal regulation and of software systems), acceptability (number of reporting regional counties), time (period from data entry at the health department to entry at the RKI) and data quality (information on immunisation status among reports of hepatitis A cases). RESULTS: For electronic processing 5 versions of Surv Net@RKI and 47 versions of the 5 commercial products are used. Additional rules of individual states expand the legal obligation for notification of the IfSG, by adding new diseases, different definitions or different reporting channels. Within the first quarter after implementation of the IfSG, 393 (90 %) of the 425 counties transmitted data weekly. The median transmission time from data entry at the health department and entry at the RKI was 5 to 7 days after the fourth reporting week. The proportion of hepatitis A case reports with information on immunisation status was 58 % (1323 of 2277); among the 1052 reports by health departments using Surv Net@RKI the proportion was 82 % (n = 858); among the 1225 reports from health departments using other programmes the proportion was 38 %. CONCLUSION: Implementation of the new surveillance system is successful. Electronic data systems should be standardised to improve data quality and simplicity. The deadlines for transmission should be shortened to allow earlier detection and control of multi state outbreaks. State-specific rules on notifiable diseases should be standardised to avoid conflicting or redundant reporting channels. PMID- 12649789 TI - [Present-day knowledge of BSE and Creutzfeldt-Jakob disease]. PMID- 12649790 TI - [A group therapy programme for eating disorders with preventive goal]. AB - Eating disorders tend to take a prolonged course because of high rates of comorbid psychiatric disorders and medical complications. We developed a cognitive behavioural group therapy programme, divided in 4 consecutively phases in a special unit. Included in the programme are therapeutic residence groups. Current and former patients are regular involved in preventive activities. PMID- 12649791 TI - [Nutritional ecology: chances of public health services to shape procedures]. AB - Nutrition ecology is the science that studies the impacts of human nutrition on the environment, on human health and on social systems. Environmental impacts are usually the result of intensive food production, food transport and food packaging; they include soil contamination and erosion, energy consumption, air contamination and waste disposal. Health impacts of contemporary western eating behavior concern nutrition-related diseases (diabetes, hypertension, gout, overweight etc) and their psychological and social consequences; nutrition ecology aims at a balanced diet and suitable eating behavior to reduce the risks of chronic disease. The social impacts of nutrition concern the effects of feed trade on poor working conditions and poverty, especially in developing countries. The strategies of nutrition ecology agree in many points with those of the agenda 21 and of Consumer Protection. As nutrition-related diseases are increasing further, the public health services in Germany discover nutrition as an important topic in health promotion and prevention. They will contribute remarkably to a sustainable development if they include the strategies of nutrition ecology in their projects. PMID- 12649792 TI - [Model project for reorganising of newborn screening]. AB - OBJECTIVE: To establish a newborn screening programme for congenital metabolic or endocrine defects which meets the national requirements in Germany: documentation of complete and early diagnosis. METHODS: The main new components of the programme consist of the introduction of a new method for detection of congenital metabolic defects (MS-Ms and 17 alpha-OHP), extension of the programme to a number of new conditions such as, fatty acid oxidation defects, aminoacidopathies, organoacidurias and AGS (adrenogenital syndrome) and the establishment of a follow-up unit of the public health service (screening centre) to allow tracking of completeness of participation rates and follow-up testing of screening positives or inconclusive screening results. RESULTS: 350 393 children were screened during 1999 to 2001. Documented participation with the screening programme could be increased from 80 % before the introduction of the new programme to at present > 98 %. The required follow-up testing for 99.1 % of the children with positive or inconclusive screening results could be achieved. In 16.5 % (1926) of the children in need of replicate testing an intervention of the screening centre was required. 271 children with congenital metabolic or endocrine defects could be identified during the 3-year period. The mean age of blood taken for screening was 3.1 days. The new screening programmes met with high acceptance from the different participants in the health care system and from parents. CONCLUSIONS: The tracking system worked excellently although the number of conditions included in the programme was high. Such a tracking system is a prerequisite for the expansion of neonatal screening programmes since many of the new conditions require early onset of treatment. PMID- 12649793 TI - [Evaluation of the premature unfitness for work among civil servants career- groups and morbidity]. AB - Out of the data obtained in a prospective study over 11,528 assessments concerning premature unfitness for work among civil servant over a period of four years the collective groups of those assessed unfit for work (n = 6,861, 2,723 women i. e. 39.7 % and 4,128 men i. e. 60.2 %) were included in this study. The median age was 54 years (women 52, men 55), 4,112 were teachers, in the administration 1,644, in the technical service 386 and in activities with special request (e. g. fire-brigade or law execution service) 483. Career groups: official-legal groups of careers in lower service categories were 266, in medium categories about 29, in elevated service 2,766 and higher service 2,146 woman officials and male officials. In all fields of activity and groups of careers psychic disorders (ICD 10: F) had a prevalence between 30.8 % in lower service and 48.0 % in the elevated service as "Premature Retirement Suffering No. 1". Women in all fields of activity were clearly more frequently concerned than men (differences from 3.1 per cent points to 18.1 per cent points middle service or technical service, p < 0.01). Second place was taken by muscular/skeletal diseases (ICD 10 group M) in 17.4 % of cases (n = 1,630), and third place by cardiovascular diseases (ICD 10 group I) in 10.4 % of cases (n = 972). The altogether lower prevalence of musculoskeletale illnesses (ICD 10: I) showed a dependency on the activity request and was highest in the technical services and in lower service categories. The median of entrance into premature unfitness for health reasons was between 15 and 9 years earlier than the age limit by law and presented itself differently in the groups of careers. Although this analysis only shows the prevalence of illnesses causal for premature retirement and not the health situation of all civil servants, it can give however pointers to emphasis for a necessary prevention. PMID- 12649794 TI - [Legal security in respect of vaccination procedures]. PMID- 12649795 TI - [Sexuality and mental retardation--love, pleasure and partnership--also for us?]. PMID- 12649796 TI - [Progress in the early diagnosis of breast carcinoma during the years 1981-1990. Results of a longitudinal study]. AB - BACKGROUND AND OBJECTIVE: Current meta-analyses have left in doubt whether general breast screening increases survival rate. This study investigated whether efforts at early diagnosis of cancer in the 1980s have had an effect on average tumor size at first diagnosis and on survival rate. PATIENTS AND METHODS: From 1981 to 1990, 1656 consecutive patients (average age 56.6 years) at the I. Women's Clinic at the Ludwig-Maximilian University of Munich and the Women's Clinic Berlin-Charlottenburg were operated on for primary breast cancer. In a retrospective analysis, average tumor size at the primary operation and survival rate were determined for two periods: 1981-1985 (n=849) and 1986-1990. Mean follow-up time was 63 months. RESULTS: There was no difference between the two cohorts regarding age (p = 0.77) and axillary node status (p = 0.14). During the follow-up period there was a gradual decrease in the tumor size at first diagnosis. (Pearson's correlation coefficient: -0.79, p < 0.001). Average tumor size in those operated on was 25 mm up to 1985, and 21 mm after 1986 (p < 0.001). Until 1985, the initial reason for mammography was the planned subsequent operation in 19% of patients (n = 164), and in 27% (n = 215; p < 0.001) since 1986. But there was no statistically significant rise in disease-specific survival rate (log rank, p=0.48). Multivariate analysis confirmed the conventional prognostic parameters, such as tumor size (relative risk 2.21) and axillary lymph node metastases (relative risk 3.57), but not the period of follow up (p=0.90). CONCLUSION: During the stated periods of follow-up there was a significant decrease in average tumor size at initial diagnosis. But this did not result in any demonstrably better disease-specific survival rate. PMID- 12649797 TI - [Human cowpox/catpox infection. A potentially unrecognized disease]. AB - HISTORY AND ADMISSION FINDINGS: A 36-year-old woman initially noticed a red spot, about pea-sized, with a central pimple over the right eyebrow and a swollen submandibular lymph node. A pressure-sensitive, 4 cm large, node developed out of this small spot, with a central, black, tightly-adhering crust bearing several varioliform vesicles around its edge. In addition to swelling of the right half of the face, the patient had a fever up to 39.5 degrees C, general malaise, nausea and vomiting. Various antibiotics were ineffective. The woman was hospitalized with a diagnosis of facial erysipelas. She owned a cat which had developed a purulent nodule on a forepaw a few days before onset of the patient's disease. LABORATORY TEST: ESR and CRP were moderately elevated, no leukocytosis and blood cultures were sterile. Wound smears showed colonization with Klebsiella pneumoniae and Enterobacter cloacae. DIAGNOSIS, TREATMENT AND COURSE: The patient's general condition improved under initially calculated antibiotic dosages, which was later adapted to the measured resistance. The black-crusted nodes became larger, however, and incision was performed on the 8 th day after hospitalization, under the suspicion of fluctuation. However, no pus was removed, but there was massive inflammatory infiltration of the soft tissue. Examination of samples of skin and part of the crust revealed orthopox virus (cowpox virus). Spontaneous healing followed within 3 weeks, leaving only a small scar. CONCLUSIONS: This was a cowpox virus in the sense of a zoonosis transmitted by the cat. In Germany, now that smallpox has been eradicated, the clinical presentation of infections with the orthopox virus, which are closely related to variola virus, are too little recognized. Atopic and immunocompromised patients are at risk of a cutaneous dissemination with a more severe course of the infectious illness; even a lethal outcome has been reported in Germany. PMID- 12649798 TI - [Pantoprazole-induced hepatitis]. AB - HISTORY: A 46-year-old woman presented for an emergency admission because of colic-like upper abdominal pain and markedly impaired general condition. Eight days before admission she had started and continued to take pantoprazole because of symptoms of gastroesophageal reflux. INVESTIGATIONS: Clinical examination and laboratory tests indicated abnormal liver functions suggesting hepatitis. Serology largely excluded an infectious, autoimmunological or metabolic cause. Duplex sonography gave no evidence of bile obstruction or Budd-Chiari syndrome. DIAGNOSIS AND COURSE: The patient's condition and laboratory tests after the drug had been discontinued gradually improved on symptomatic treatment, indicating pantoprazole-induced hepatitis. CONCLUSION: Intake of proton pump inhibitors is a rare cause but should be considered in the differential diagnosis of hepatitis of uncertain etiology. PMID- 12649799 TI - [Therapy of adrenal cortex insufficiency]. PMID- 12649800 TI - [Patient satisfaction with inpatient care. Evaluation, methods and special features]. PMID- 12649801 TI - [The heart of the athlete]. PMID- 12649802 TI - [Passing on opened drug packages by the doctor to patients. Decision of the Detmold District Court on February 16, 2001--24OWi 22 Js 778/00]. PMID- 12649803 TI - [What should follow a treatment for pruritus in thrombocytosis?]. PMID- 12649804 TI - [Is systemic administration of N-acetylcysteine effective?]. PMID- 12649805 TI - [The "needle-man" (more than 40,000 injections in 40 years): calcification of the basal ganglia in hypoparathyroidism]. PMID- 12649806 TI - [Diabetic ketoacidosis and hyperosmolar hyperglycemia. 24 consecutive cases]. PMID- 12649807 TI - Genome architecture catalyzes nonrecurrent chromosomal rearrangements. AB - To investigate the potential involvement of genome architecture in nonrecurrent chromosome rearrangements, we analyzed the breakpoints of eight translocations and 18 unusual-sized deletions involving human proximal 17p. Surprisingly, we found that many deletion breakpoints occurred in low-copy repeats (LCRs); 13 were associated with novel large LCR17p structures, and 2 mapped within an LCR sequence (middle SMS-REP) within the Smith-Magenis syndrome (SMS) common deletion. Three translocation breakpoints involving 17p11 were found to be located within the centromeric alpha-satellite sequence D17Z1, three within a pericentromeric segment, and one at the distal SMS-REP. Remarkably, our analysis reveals that LCRs constitute >23% of the analyzed genome sequence in proximal 17p -an experimental observation two- to fourfold higher than predictions based on virtual analysis of the genome. Our data demonstrate that higher-order genomic architecture involving LCRs plays a significant role not only in recurrent chromosome rearrangements but also in translocations and unusual-sized deletions involving 17p. PMID- 12649808 TI - Missense mutations in the homeodomain of HOXD13 are associated with brachydactyly types D and E. AB - HOXD13, the most 5' gene of the HOXD cluster, encodes a homeodomain transcription factor with important functions in limb patterning and growth. Heterozygous mutations of human HOXD13, encoding polyalanine expansions or frameshifts, are believed to act by dominant negative or haploinsufficiency mechanisms and are predominantly associated with synpolydactyly phenotypes. Here, we describe two mutations of HOXD13 (923C-->G encoding Ser308Cys and 940A-->C encoding Ile314Leu) that cause missense substitutions within the homeodomain. Both are associated with distinctive limb phenotypes in which brachydactyly of specific metacarpals, metatarsals, and phalangeal bones is the most constant feature, exhibiting overlap with brachydactyly types D and E. We investigated the binding of synthetic mutant proteins to double-stranded DNA targets in vitro. No consistent differences were found for the Ser308Cys mutation compared with the wild type, but the Ile314Leu mutation (which resides at the 47th position of the homeodomain) exhibited increased affinity for a target containing the core recognition sequence 5'-TTAC-3' but decreased affinity for a 5'-TTAT-3' target. Molecular modeling of the Ile314Leu mutation indicates that this mixed gain and loss of affinity may be accounted for by the relative positions of methyl groups in the amino acid side chain and target base. PMID- 12649809 TI - Conditional JAG1 mutation shows the developing heart is more sensitive than developing liver to JAG1 dosage. AB - Mutations of Jagged 1 (JAG1), a ligand in the Notch signaling pathway, cause Alagille syndrome (AGS). AGS is an autosomal dominant, multisystem disorder with variable expressivity, characterized by bile duct paucity and resultant liver disease in combination with cardiac, ocular, skeletal, and facial findings. JAG1 mutations in AGS include gene deletions and protein truncating, splicing, and missense mutations, suggesting that haploinsufficiency is the mechanism of disease causation. With limited exceptions, there is no genotype-phenotype correlation. We have studied a JAG1 missense mutation (JAG1-G274D) that was previously identified in 13 individuals from an extended family with cardiac defects of the type seen in patients with AGS (e.g., peripheral pulmonic stenosis and tetralogy of Fallot) in the absence of liver dysfunction. Our data indicate that this mutation is "leaky." Two populations of proteins are produced from this allele. One population is abnormally glycosylated and is retained intracellularly rather than being transported to the cell surface. A second population is normally glycosylated and is transported to the cell surface, where it is able to signal to the Notch receptor. The JAG1-G274D protein is temperature sensitive, with more abnormally glycosylated (and nonfunctional) molecules produced at higher temperatures. Carriers of this mutation therefore have >50% but <100% of the normal concentration of JAG1 molecules on the cell surface. The cardiac specific phenotype associated with this mutation suggests that the developing heart is more sensitive than the developing liver to decreased dosage of JAG1. PMID- 12649810 TI - Renewal. PMID- 12649811 TI - Cultural diversity: love's labor lost: success and failure in global nursing. PMID- 12649812 TI - Education: the best of times could be now. PMID- 12649813 TI - Legal and ethical issues: the right way to live. PMID- 12649814 TI - Evaluation of the Penn Macy initiative to advance academic nursing practice. AB - In the realm of academic practice, schools of nursing, especially those in research-intensive environments, face a dilemma. These schools are in the best position to shape nursing science through research based in academic practice. Yet they also face formidable challenges to maintaining balance among the three legs of the tripartite mission. The questions of whether and how to commit resources to move forward their academic practice agendas are not answered easily. The Penn Macy Initiative to Advance Academic Nursing Practice aimed to assist such schools in their development of academic practices through a week long intensive institute and 1 year of consultation and follow-up. An evaluation of the outcomes of that experience for 21 participant schools together with the baseline characteristics that appeared, retrospectively, to be associated with success are presented and discussed. PMID- 12649815 TI - Teaching, practice, and research: an integrative approach benefiting students and faculty. AB - A project showing the integration of teaching, practice, and research is described. The project involved using information gathered as part of baccalaureate nursing students' assessments of women's perceptions of and responses to cesarean birth to extend a program of nursing research. Benefits to student learning and faculty research programs are identified. The project serves as a prototype for the teaching, practice, and research activities of all students and faculty in nursing and other professional disciplines. PMID- 12649816 TI - Minority international research training program: global collaboration in nursing research. AB - In the current century, nurses, along with other health professionals, require preparation for their roles in international health. For faculty and students to practice and learn in this global community, they also will want to have added knowledge of economics, business, and public policy. To gain an immersion experience in global health, nursing faculty and students will need to spend time abroad collaborating with nursing colleagues in other countries. This report describes an academic-research training program (Minority International Research Program [MIRT]) established 7 years ago to enhance the international health experience of minority nursing students. Qualified undergraduate and graduate nursing students are recruited and paired with faculty mentors to conduct short term research abroad for a period of 10 to 14 weeks. The purpose of international research experiences for minority nursing students is to develop leaders in nursing science and to increase collaboration in the resolution of global health issues. To date, 26 undergraduate students, 22 graduate students, 6 postdoctoral trainees, and 11 faculty mentors have participated and completed the program through research immersion experiences in 9 countries. The program is described in terms of its strengths, challenges, and lessons learned, as well as opportunities for future activities. PMID- 12649817 TI - Preceptor leadership style and the nursing practicum. AB - Preceptors are clinically based nurses that agree to participate with nursing students in an effort to provide them with opportunities to reinforce their knowledge with clinical experience. Preceptors serve as nursing role models to students and can facilitate their understanding and socialization into the reality of nursing. Studies on preceptorships have focused traditionally on the experience of precepting from the perspectives of student, employer, and preceptor. Literature related to methods for matching preceptors and students is very limited. Selection of preceptors to serve as role models for students should not be limited to clinical skill alone. Student learning and clinical experience can be influenced directly by the leadership characteristics that a preceptor possesses. To make the experience of precepting positive and beneficial for preceptor, student, and faculty, an examination and discussion of the potential preceptor's leadership style should be conducted. Situational leadership and the four different styles of leadership identified by Hersey and Blanchard provide a useful model for identifying leadership traits in preceptors that nursing faculty will find useful as they seek to match preceptors in a way that will facilitate knowledge acquisition and application for nursing students in the clinical setting. PMID- 12649818 TI - Evaluation of an informal peer group experience on baccalaureate nursing students' emotional well-being and professional socialization. AB - Although studies have documented the importance of the academic environment in promoting positive outcomes among students, few quantitative studies in nursing have been performed to identify strategies through which a nurturing learning environment can be created. A randomized two-group pretest-posttest design was used to investigate effects of an informal peer group experience on baccalaureate nursing students' emotional well-being and professional socialization as caring practitioners. Groups did not differ significantly on the outcomes measured in this study. As a whole, students showed statistically significant increases in anxiety, depression, and negative affect, along with decreased self-esteem and positive affect during the junior year of nursing school. Psychological problems and stress-related symptoms have been well documented among college students in general and professional students in particular. Although the findings from this study are not unique, they suggest the need to reconsider strategies by which the affective and professional socialization goals of undergraduate nursing education can be achieved. PMID- 12649819 TI - A picture of a work of art as an empathy teaching strategy in nurse education complementary to theoretical knowledge. AB - A randomized study was performed to investigate whether a teaching intervention program using a reproduction of Edvard Munch's painting, "The Sick Girl," stimulated student nurses to engage in learning about empathy. A control group was used to control for effects of the visual art dialogues in the intervention group. The study was undertaken at a university college of health science in Sweden. Data were collected from student nurses (n = 144) during their first study year. The result showed a significant improvement in the visual art group compared with the matched control group. Students in the visual art group were more engaged in learning about empathy when measured with the Wheel Questionnaire parameters of structure, motivation, and emotional investment. The present study supported the idea that Edvard Munch's painting could be used in nurse education complementary to theoretical knowledge on empathy to stimulate student nurses to engage in empathy learning. PMID- 12649820 TI - Nontuberculous mycobacterial adenitis in children: diagnostic and therapeutic management. AB - We reviewed a series of 45 patients affected by nontuberculous mycobacterial adenitis of the neck observed in the Ear, Nose, and Throat Institute of S. Orsola Malpighi Hospital-Bologna over a 20-year period between 1981 and 2001. The mean age was 5.5 years. Patients were tested by using the differential Mantoux test, which was the principal diagnostic tool in the case of atypical mycobacterial infections. Forty-two patients were surgically treated by total excision of infected nodes, whereas parotidectomy with sparing of facial nerve was performed in those 3 cases with intraparotid nodes involvement. In all cases, the histopathological diagnosis was tubercular granulomatous lymphadenitis. The culture growth of nontuberculous mycobacteria was positive in 13 cases with a marked prevalence of the avium-intracellular germs. The disease was eradicated in all patients. The diagnostic and therapeutic management of nontuberculous mycobacterial adenitis is discussed in this retrospective study. PMID- 12649821 TI - Revisiting a controversial surgical technique in the treatment of bulimic parotid hypertrophy. AB - Bulimia nervosa is a recognized cause of bilateral parotid gland enlargement that may not be appreciated by most otolaryngologists. Classically the parotid enlargement is transient but may be permanent and spectacular. Our review of the published literature revealed only 2 previous cases that have been considered for superficial parotidectomy, along with much debate as to the efficacy of such treatments. Neither case reported any postoperative complications. We present a bulimic patient who developed bilateral postoperative fistulae after surgery for marked, permanent enlargement of her parotids. Both fistulas closed spontaneously. However, a year later she has relapsed into bulimic behavior and developed further parotid gland hypertrophy. Because of this complication, we believe surgery has no role to play in the overall treatment of these severe cases unless the patients show a prolonged cessation of their bulimic behavior. PMID- 12649822 TI - A foreign body in the pharynx migrating through the internal jugular vein. AB - We present an unusual and a rare case study of a 45-year-old woman who had swallowed a sharp pointed metallic foreign body while eating meat. The foreign body had migrated from the cricopharynx through the parapharyngeal space and penetrated the internal jugular vein over a period of 10 days presenting as a tender neck swelling. The management of this case is discussed here in brief. PMID- 12649823 TI - Aberrant carotid artery in the middle ear: multislice CT imaging aids in diagnosis. PMID- 12649824 TI - Health-Related Quality of Life in rhino surgery. AB - BACKGROUND: Health-Related Quality of Life (HRQL) measures the impact of a pathologic condition on patient's daily life. Besides from disease-related symptoms, it includes a wide spectrum of daily life activities such as physical and social activities, emotional problems, general feeling, and so on. HRQL of patients with nasal diseases is known to correlate only moderately to conventional clinical markers. HRQL data about patients undergoing rhino surgery are not available to date. OBJECTIVE: The purpose of this study was to study HRQL in patients undergoing nasal surgery and to measure therapeutic effects 3 months postoperatively. METHODS: We used a specific health profile HRQL questionnaire with 25 items summarized in 6 symptom groups: sleep; nonnasal, nasal, and emotional symptoms; headache; and practical problems (symptom score 1 to 4). A visual analog scale (0 to 10) was given to measure the patient's general feeling related to their nasal disease. One hundred eighty-one patients undergoing nasal surgery for various reasons were included preoperatively. One hundred seven of them could be interviewed 3 months postoperatively to study therapeutic effects of our surgical intervention. RESULTS: Patient's pre- and postoperative HRQL status could be determined, and differences in disease-related subgroups could be identified. Comparing pre- and postoperatively gained HRQL data revealed a significant improvement in symptom score of 23 of 25 items and according to the following in all symptom groups: sleep preoperatively 2.46 versus postoperatively 1.98 (P <.01), nonnasal symptoms preoperatively 2.13 versus postoperatively 1.91 (P <.01), headache preoperatively 2.17 versus postoperatively 1.72 (P <.01), practical problems preoperatively 2.47 versus postoperatively 2.06 (P <.01), nasal symptoms preoperatively 2.39 versus postoperatively 1.90 (P <.01), and emotional problems preoperatively 2.01 versus postoperatively 1.81 (P <.01). The general feeling score improved from preoperatively 6.47 to postoperatively 3.95 (P <.01) as well. CONCLUSIONS: We could measure patient's HRQL status pre- and postoperatively, could work out peculiarities of the studied subgroups, and showed therapeutic benefits of our surgical intervention. PMID- 12649825 TI - Retrospective analysis of Ki-67 antigen expression in paraffin tissue blocks of laryngeal squamous cell carcinoma. AB - The level of Ki-67 antigen expression has been proven to correlate with cellular proliferation. The aim of the study was to show correlation of Ki-67 expression with the clinical stage and its predictive value for the prognosis of laryngeal carcinoma. The study included 48 patients with surgically removed carcinoma of the larynx. Study results pointed to a significant difference in the mean value of Ki-67 reactivity between the study and control group consisting of cadaveric laryngeal tissue free from the disease. Correlation was found between the TNM stage of carcinoma and percentage of Ki-67-labeled cells. A significant correlation was also observed between Ki-67 reactivity at the time of laryngeal surgery and 2-year clinical outcome of the disease. Ki-67 reactivity was significantly higher in the group of patients with disease remission compared with the control group but was significantly lower as compared with the group of patients with poor outcome. PMID- 12649826 TI - Characterizations of life-threatening deep cervical space infections: a review of one hundred ninety-six cases. AB - PURPOSE: The incidence of deep cervical space infections has decreased after the introduction of antibiotics and improvement of oral hygiene, but they still may be lethal especially when life-threatening complications occur. In this article, we try to find out whether there are predisposing factors related to complicated deep cervical space infections and prolonged hospitalization. MATERIALS AND METHODS: We conducted a retrospective analysis of medical records of 196 patients with deep neck infections during the period from March 1996 to February 2002. Among the 196 patients, 15 patients developed lethal complications. As for data analysis, multiple regression and logistic regression with dummy variable were used. RESULTS: In multiple regressions, patients with older age or with neck swelling, trismus, underlying diseases, complications, and C-reactive protein more than 100 microg/mL stayed longer in the hospital, and the P value reached statistical significance (P <.05). In logistic regression, male patients and patients with neck pain had negative correlation, and the P value reached statistical significance (P <.05). Patients with neck swelling and patients with respiratory difficulty had positive correlation, and the P value reached statistical significance (P <.05). That means female patients, patients with neck swelling, and patients with respiratory difficulty were more likely to have complicated deep neck infections. CONCLUSION: Complicated deep neck infections remain potentially fatal, but the morbidity and mortality can be reduced. Doctors should pay more attention to those high-risk patients; they are female patients, patients with neck swelling, and patients with respiratory difficulty. PMID- 12649827 TI - An unusual cause of foreign-body sensation in the throat: corniculate cartilage subluxation. AB - Corniculate cartilage subluxation is a pathology that has not been reported in the medical literature. Causing vague pharyngeal symptoms, this disease entity may be overlooked or misdiagnosed as chronic nonspecific pharyngitis, globus pharyngis, or laryngopharyngeal reflux disease. A careful laryngoscopic examination is the key to detect this disorder. In this article, we present a 45 year-old patient with the complaints of foreign-body sensation in the throat, difficulty in swallowing solid food, and an urge to turn his head toward the left during the act of swallowing. His laryngeal examination revealed corniculate cartilage subluxation, and excision of the subluxated cartilage was performed by microlaryngoscopic surgery. The patient was free of his symptoms immediately after the operation and remained as such in the 6 months of follow-up. PMID- 12649828 TI - Mixed myxoid/round cell liposarcoma of the scalp. AB - Liposarcoma of the head and neck is rare. Only 12 cases of scalp liposarcoma have been previously reported. In this report, we describe a case of myxoid/round cell liposarcoma in the scalp of a 28-year-old woman. This case report highlights a histologic pattern rarely reported in the head and neck but consistent with the evolving classification of liposarcomas. PMID- 12649829 TI - Rapidly progressive bilateral sensory neural hearing loss as a presentation of mitochondrial neurogastrointestinal encephalomyopathy. PMID- 12649830 TI - [Hemophagocytic syndrome]. PMID- 12649831 TI - [Trends of invasive fungal infections in hematological malignancies and new antifungal agents]. PMID- 12649832 TI - [A long-lasting idiopathic factor V inhibitor]. AB - A rare case of a long-standing idiopathic acquired blood coagulation factor V (FV) inhibitor is reported. A 78 year-old male was admitted complaining of hematuria and tarry stools of two weeks' duration. He was noted to have a prolonged prothrombin time and activated partial thromboplastin time. A mixing study suggested the existence of an inhibitor and the activity of FV was less than 1%. Western blot analysis revealed the presence of anti-FV antibodies in the patient's serum. Because of significant bleeding tendencies, the patient received fresh-frozen plasma, platelet transfusions and methylprednisolone. His PT, APTT and FV activity were partially corrected although the disease was exacerbated when the steroid dose was tapered off. The patient was then treated with low dose cyclophosphamide. The majority of the cases with acquired FV inhibitor occur spontaneously in older patients but this condition may be associated with a surgical procedure, administration of antibiotics, tuberculosis or malignant diseases; some of them are, however, idiopathic. No underlying disease was detected in our present patient. The inhibitor disappears within 10 weeks in most patients and the prolonged presence of an idiopathic FV inhibitor is rare. PMID- 12649833 TI - [Simultaneous occurrence of acute myeloid leukemia with multilineage dysplasia and multiple myeloma]. AB - An 82-year-old woman was admitted to our hospital because of dizziness and petechiae. Peripheral blood examination showed severe anemia and thrombocytopenia. Bone marrow aspiration revealed 42% leukemic blasts positive for peroxidase with multilineage dysplasia, leading to a diagnosis of acute myeloid leukemia with multilineage dysplasia. The levels of the patient's marrow plasma cells increased to 12%, whereas serum levels of IgG, A, and M dropped. lambda type Bence Jones protein was detected on urine immunoelectrophoresis. The total urinary protein was 3, 960 mg/day. Bone scintigraphy detected multifocal uptake in the ribs. The diagnosis was multiple myeloma developing simultaneously with acute myeloid leukemia. Possible mechanisms for the occurrence of acute myeloid leukemia and multiple myeloma were discussed. PMID- 12649834 TI - [Lymphoblastic lymphoma presenting as a solitary subcutaneous mass]. AB - A solitary cutaneous or subcutaneous mass in lymphoblastic lymphoma (LBL) is a rare manifestation. A 15-year-old girl presented with a subcutaneous LBL on her left back. There were no other lesions. Complete remission (CR) was achieved after 2 courses of ACOMP-B (doxorubicin, cyclophosphamide, vincristine, methotrexate, prednisolone, and bleomycin) therapy. Two courses of the chemotherapy and 4 sessions of prophylactic intrathecal methotrexate (15 mg/body) and hydrocortisone (25 mg/body) were added after CR. However, the patient relapsed with bone marrow involvement after 14 months remission. It is necessary to accumulate more experience with this kind of case to find the appropriate treatment strategy for solitary cutaneous or subcutaneous LBL. PMID- 12649835 TI - [Clinical significance of anti-phosphatidylserine-prothrombin complex antibodies in the diagnosis of anti-phospholipid syndrome]. AB - In this study, we clarified the clinical significance of IgG anti phosphatidylserine-prothrombin complex (PS-PT) antibodies in the diagnosis of antiphospholipid syndrome (APS). The study population consisted of 113 patients with SLE and lupus-like disease. IgG anti-PS-PT antibodies were examined by ELISA. These antibodies were detected in 31% of the population, and were significantly associated with thrombosis and fetal loss. The frequency was significantly higher in patients who did not satisfy the Sapporo criteria, but met the Harris and Hughes diagnostic criteria (36%) than in non-APS patients (14%). The above findings suggest that IgG anti-PS-PT antibodies should be listed in the diagnostic criteria for APS. PMID- 12649836 TI - [Idiopathic thrombocytopenic purpura associated with mantle cell lymphoma showing an increase in the platelet count receiving anti-CD20 antibody rituximab]. AB - A 68-year-old man was admitted for bleeding tendency and generalized lymphadenopathy. A diagnosis of idiopathic thrombocytopenic purpura (ITP) associated with mantle cell lymphoma was made. The anti-CD20 monoclonal antibody rituximab 375 mg/m2 was given intravenously once weekly for four consecutive weeks. The patient's platelet counts increased gradually from 0.6 x 10(4)/microliter to 5.8 x 10(4)/microliter. At eighteen weeks after discontinuation of rituximab medication, his platelet count increased again to 10. 3 x 10(4)/microliter and this value has been sustained up to the time of writing. This suggests that rituximab is useful in the treatment of ITP associated with malignant lymphoma. PMID- 12649837 TI - [Mild cognitive impairment: current update]. PMID- 12649838 TI - [Therapeutic angiogenesis by bone marrow cells]. PMID- 12649839 TI - [Changes in age-associated neurodegenerative diseases in the half century]. PMID- 12649840 TI - [CGA and QOL in community-dwelling elderly]. PMID- 12649841 TI - [Biomarkers, mild cognitive impairment and early diagnosis of Alzheimer's disease]. AB - Elderly people are concerned about changes in their cognitive functioning. Since cholinergic therapies for Alzheimer's disease have been developed and become widely accepted, elderly people have come to visit clinics to seek medical advice about whether such a subtle change in cognitive ability may represent an early manifestation of Alzheimer's disease (AD). If they are likely to develop dementia or AD, they want to receive immediate medical treatment as soon as possible to prevent further loss of cognitive functioning so that they can live independently as long as possible. The first priority in the clinical application of a biomarker is that biomarker should contribute to early diagnosis of dementia. Among such biomarkers, we believe that cerebrospinal fluid markers and functional brain imaging are clinically the most applicable procedures. Since 1993, we have collected 623 cerebrospinal fluid (CSF) samples at The Tohoku University Hospital for evaluation of dementia (age: 42-93). We found that CSF/phospho-tau measures produced the most adequate sensitivity (85.2%) and specificity (85.0%) in the diagnosis of AD as a sole bio-marker. The CSF levels of A beta 1-42 showed a strong positive correlation with the Mini-mental state examination score and brain glucose metabolism by positron emission tomography. The baseline levels of both total-tau and phospho-tau in CSF increased in approximately 70% of patients with mild cognitive impairment who later developed AD, suggesting that pathological change in the brain might start years before dementia becomes clinically manifested. A combined use of CSF-tau and IMP-SPECT improved the predictability of the transition from mild cognitive impairment into AD. PMID- 12649842 TI - [Development of animal models for therapy of Alzheimer's disease]. PMID- 12649843 TI - [Common disease and dementia]. PMID- 12649844 TI - [Neurofibrillary tangle formation and Alzheimer's disease with brain aging]. PMID- 12649845 TI - [Neuronal cell death by Alzheimer's disease-relevant insults and its rescue]. AB - Neuronal cell death accounts for the clinical manifestations in Alzheimer's disease (AD). To establish the curative therapy of AD, neuroprotection is one of the primary therapeutic targets, and the elucidation of the mechanism of neuronal cell death is mandatory. Detailed characterization of neuronal cell death caused by familial AD (FAD)-linked mutant genes revealed that different cell death pathways are evoked by different types of mutants. Humanin (HN), a newly identified neuroprotective peptide, suppresses neuronal cell death caused by all known FAD mutants and A beta, while it has no effect on neuronal cell death caused by AD-irrelevant insults. The functional target of HN is the antagonism to neuronal death, not the modulation of A beta production, suggesting that HN-based medication can be combined with other remedies targeting A beta. HN is a promising seed for a novel therapy aiming at complete cure of AD through the suppression of neuronal loss. PMID- 12649846 TI - [Acceptance of dialysis therapy in elderly patients with chronic renal failure]. AB - The aim of the present study is to clarify relevant factors concerning acceptance of dialysis therapy in elderly patients with chronic renal failure. Patients with advanced renal failure aged 60 years and over (152 cases) were investigated. The male/female ratio was 85:67. The age was 76 +/- 7 years (mean +/- standard deviation). The proportion of patients with acceptance of dialysis to patients with non-acceptance was 121:31. In all patients, the cause of renal failure (non diabetes/diabetes), serum albumin level, comorbid conditions, ambulation, cognitive function, marital status, and presence of younger cohabitants were surveyed. The patients were divided into two groups for each category. Patients were categorized as the young-old (aged 60 to 74 years) and the old-old (aged 75 years and over). Serum albumin level was categorized as either low (less than 3.5 g/dl) or normal (3.5 g/dl and over). The number of patients who accepted dialysis therapy was evaluated for each group. Intergroup comparisons were carried out by the chi 2 test. Statistically significant factors were age (p < 0.0001), serum albumin level (p = 0.016), ambulation (p = 0.011), cognitive function (p < 0.0001), and marital status (p = 0.009). Multivariant logistic regression analysis was also performed using background factors as explanatory variables and acceptance or non-acceptance of dialysis therapy as a dependent variable. The factors presented by the nominal scale were converted to dummy variables. Statistically significant factors were age (p < 0.0001) and cognitive function (p < 0.0001). Serum albumin level, ambulation, and marital status were significant only in the chi 2 test. This could be explained by the close correlations of these factors with age and cognitive function. The old-old category and poor cognitive function were dominant factors with regard to non-acceptance of dialysis therapy. PMID- 12649847 TI - [Intervention study using a fall prevention program to prevent functional decline of old-old elderly in a rural community]. AB - The purpose of this study was to determine the effects of a fall prevention program for old-old elderly in a rural community in Japan. The subjects were 71 inhabitants aged 75 and over who used community health and welfare services such as day services and social activity classes. They were divided into two groups: 34 in the experimental group, and 37 controls. Physical therapists and care staff instructed the experimental group in fall prevention exercises once per two weeks from December 2000 to March 2001. The control group utilized the usual services without any special instructions. All subjects were evaluated by physical performance tests and a self-administered questionnaire before and after 4 months following the intervention. Collected data were analyzed by two-way ANOVA using intervention and time as independent variables, and physical performance tests as dependent variables. Twenty-three of the experimental group and 26 of the control group completed both baseline and follow-up surveys. There were no significant differences in physical ability between the two groups at the time of the baseline survey. The experimental group showed excellent compliance and participated in all sessions during the intervention period. Interaction effects on body mass index and Timed Up and Go Test were shown to be significant (F = 5.623, P < 0.01: F = 6.541, P < 0.05). There were no changes in terms of other physical and psychological factors. Our results showed specific exercises could improve some aspects of physical performance. Since the group exercise program can be effective for old-old elderly, it might contribute to decreasing falls and prolong independent living. PMID- 12649848 TI - [Medical students' understanding of geriatric medicine]. AB - With the rapid aging of Japan's population, medical professionals who specialize in geriatric medicine are in unprecedented demand. At Ehime University School of Medicine, the Department of Geriatric Medicine was established on April 1,1997. To assess the status of geriatric medicine in undergraduate education, we surveyed the opinions of medical students on geriatric medicine. A questionnaire was sent to both fourth-year medical students before their classes on internal medicine and geriatric medicine had ended and to sixth-year medical students after their classes had already ended. Medical students of Ehime University School of Medicine had significantly more interest in geriatric medicine than students at other medical schools. From a clinical viewpoint of problems in old age, they considered that cerebrovascular disease, cardiovascular disorders, neurocognitive disorders and pulmonary disease were important. A total of 60% agreed that all medical schools should have classes in geriatric medicine, which is a higher rate than that of previous reports. To study the opinions of medical students on textbooks on geriatrics, we also conducted a questionnaire among medical students. Although 90% of students had their own textbooks of internal medicine and almost half (42%) had textbooks on physical examination, none had their own textbook of geriatric medicine. The most frequent reason was "hard to choose the best textbook". The present study indicates that to develop education in geriatric medicine, it is important to make a recommendable text book for medical students, which is easily understood with high quality and originality. PMID- 12649849 TI - [Five cases of acute hemorrhagic rectal ulcer in aged patients]. AB - We report our experience with five cases of acute hemorrhagic rectal ulcer in aged patients. The patients included two men and three women whose ages ranged from 73 to 86 with a mean of 79.6. All cases had underlying disorders, including cerebral vascular disease, fracture of the fibula, senile dementia, chronic rheumatoid arthritis, or were recuperating from a cardiovascular operation. All cases were in bed rest and complained of sudden anal bleeding without pain. Endoscopic examination showed rectal ulcer localized in the lower rectum close to the dentate line. Four cases were treated by HSE injection and ligation by clipping devices with endoscopy and one case was surgically ligated by a transanal approach. All cases had good outcomes. Emergency colonoscopic examination is considered essential for the diagnosis and treatment of acute hemorrhagic rectal ulcers, especially in aged patients with severe underlying disorders. PMID- 12649850 TI - [Beneficial effect of cibenzoline in a patient with hypertrophic obstructive cardiomyopathy complicated with idiopathic interstitial pneumonia]. AB - A 72-year-old woman who had idiopathic interstitial pneumonia was admitted due to general fatigue. Echocardiography revealed asymmetric septal hypertrophy and systolic anterior movement of the mitral valve. In addition, Doppler echocardiography revealed a pressure gradient of 52 mmHg in the left ventricular outflow tract. Hypertrophic obstructive cardiomyopathy was diagnosed. Because she had a respiratory disease, she was treated with cibenzoline instead of beta blockers. After treatment her pressure gradient decreased to 10 mmHg, but respiratory symptom remained unchanged. This finding suggests that cibenzoline is useful for patients with hypertrophic obstructive cardiomyopathy complicated with respiratory disease. PMID- 12649851 TI - [A case of primary hepatic lymphoma successfully treated by THP-COP therapy]. AB - A 65-year-old woman was admitted due to epigastralgia. The presence of a tumor mass in the umbilical portion of the liver was recognized by abdominal ultrasonography and computed tomography scan. Needle biopsy of the tumor showed non Hodgkin's lymphoma (diffuse large B cell type) by histology and histoimmunology. She was treated by the THP-COP protocol, and complete remission was achieved after the third cycle. Primary hepatic lymphoma is so infrequent that standard treatments are not established yet. Most cases of primary hepatic lymphoma are treated by surgical resection in Japan; however our conservative approach to remission is considered as very helpful for discussing how to treat primary hepatic lymphoma. PMID- 12649852 TI - [Study of digestive diseases in post-genome era]. PMID- 12649853 TI - [Matrix metalloproteinases; the past, present, and future]. PMID- 12649854 TI - [Matrix metalloproteinases in human gastrointestinal carcinomas]. PMID- 12649855 TI - [A case of jejunal arteriovenous malformation diagnosed with the aid of intraoperative endoscopy]. PMID- 12649856 TI - [A case of hemangioma of the small intestine in which the region could be diagnosed preoperatively]. PMID- 12649857 TI - [A case of gastrointestinal stromal tumor of the duodenum]. PMID- 12649858 TI - [An autopsy case of hepatocellular carcinoma with a solitary mediastinal lymph node metastasis]. PMID- 12649859 TI - [A case of spontaneous biloma complicated with cholelithiasis and cholecystitis]. PMID- 12649860 TI - [A case of small cell carcinoma of the extrahepatic bile duct which could be diagnosed before operation]. PMID- 12649861 TI - [A case of spleen rupture in chronic pancreatitis]. PMID- 12649862 TI - [A case of recurrent tumor-forming pancreatitis effectively treated with steroid]. PMID- 12649863 TI - [A case of primary abscess of the greater omentum]. PMID- 12649864 TI - [A case of pneumatosis cystoides intestinalis with diabetes mellitus successfully treated by hyperbaric oxygen therapy]. PMID- 12649865 TI - [Perioperative cerebral ischemia and the possibility of neuroprotection by inhalational anesthetics]. AB - Deteriorated neurological outcome is not rare after major surgeries. With aging of the surgical population in Japan, an increasing frequency of perioperative cerebral ischemia is expected. To avoid neurological deterioration after surgery and general anesthesia, especially in high-risk patients, it is important to provide appropriate CNS-oriented anesthesia management. Controlling factors such as cerebral perfusion pressure, arterial blood gases, blood glucose concentration and body temperature may influence brain metabolism and neurological outcome. Inhalational anesthetics may also play an important role in perioperative neurological management, because numerous studies have demonstrated some degree of neuroprotective effect with halothane, sevoflurane, desflurane and isoflurane. Among these inhalational anesthetics, isoflurane has been most extensively studied regarding neuroprotection against cerebral ischemic insult. Although no clinical outcome trials have been performed, both in vivo and in vitro studies have consistently shown that the isoflurane provides neuroprotection. However, it is also suggested that factors such as sympathetic reactivity, brain temperature, anesthetic dosage, timing of anesthetic administration, and co-administration of nitrous oxide might affect the neuroprotective effect of isoflurane. PMID- 12649866 TI - [Diagnosis of the autonomic nervous function during sedation by peripheral blood flow and psychogenic perspiration]. AB - BACKGROUND: Pain induces a variety of physiological responses, many of which are mediated by the sympathetic nervous system. Among these are a reduction in peripheral blood flow and evaporative cutaneous water loss (sweating). We therefore tested the hypothesis that adequate sedation obliterates the normal pain-induced reduction in peripheral blood flow and an increase in evaporative water loss. METHODS: We studied eight volunteers. Two different painful stimuli were randomly applied: 1) electrical pulp stimulation (200 microamperes) and 2) electrical pain stimulation on the right upper thigh (80 mA). Conscious sedating was controlled by propofol infusion titrated to a Bispectral Index near 80, or near 60. RESULTS: At each stimulation, peripheral blood flow detected by laser Doppler decreased without any relation to the level of consciousness (by Bispectral Index). On the other hand, although the psychogenic perspiration rate increased significantly at alert level, during BIS 80 or 60 level, the increase was not significant. CONCLUSION: Peripheral blood flow reacts most to pain stimulation during intravenous sedation. PMID- 12649867 TI - [Propofol depresses the activity of hypoglossal nerve more than that of phrenic nerve in rabbits]. AB - BACKGROUND: Respiratory depression, such as obstruction of upper airway and inadequate ventilation, often appears during sedation and anesthesia. We studied the effect of propofol on the upper airway patency and the inspiratory drives. METHODS: Experiments were performed on the adult rabbits vagotomized, paralyzed and ventilated with nitrous oxide-oxygen-sevoflurane. We evaluated and compared depressant effect of propofol on the peak amplitude of both hypoglossal nerve (AMP-HG) and phrenic nerve (AMP-PH) activities, inspiratory time (Ti), expiratory time (Te) and respiratory cycle (Tc). RESULTS: Bolus injections of propofol transiently reduced AMP-HG more than AMP-PH (18 and 70% of control, respectively). But AMPs returned to the control levels about 10-15 min after the injection. For 0.5 mg.kg-1.min-1 continuous infusion, propofol soon began to reduce both AMPs. AMP-HG was reduced to about 20% and AMP-PH to 60% of control. Administration of propofol with 1.0 mg.kg-1.min-1 caused more reduction in AMPs with respiratory slowing and AMP-HG disappeared in some animals. CONCLUSION: During the sedation with low dose of propofol, we need to pay attention to potential upper airway obstruction. In addition to the above, high doses of propofol could reduce spontaneous inspiratory drive, and we need to keep both upper airway patency and sufficient ventilation. PMID- 12649868 TI - [Pulmonary embolism in three surgical patients despite prophylactic measures]. AB - Pulmonary embolism after surgery became a major problem in Japan recently. From the year 2000, we started the prophylaxis against pulmonary embolism in orthopedic or gynecological surgical patients with some risk factors of pulmonary embolism. But we experienced three cases of pulmonary embolism although two of them had received subcutaneous heparin after surgery. We consider that prophylactic measures are recommended for the patient with risk factors of pulmonary embolism. Pulmonary embolism should be considered when surgical patients complain of abnormality such as chest pain. PMID- 12649869 TI - [Anesthesia for 15 morbidly obese parturients undergoing cesarean section]. AB - From 1994 to June 2001, 15 parturients weighing more than 100 kg underwent cesarean section at our institution. Their medical and anesthetic records were reviewed retrospectively. They consisted of 10 primiparous and 5 multiparous patients. Mean body weight was 108.4 +/- 6.3 kg (mean +/- SD) and body mass index (weight in kilograms/[height in meters]2) was 41.6 +/- 2.8 kg.m-2 (min 36.6, max 49.7). Maternal complications included preeclampsia (n = 7, 46.7%), diabetes mellitus (n = 6, 40%) and asthma (n = 1). Of 15 patients, cesarean section was performed under spinal anesthesia in 11 patients and under epidural anesthesia in 4, and none received general anesthesia. All the patients gave live births. Intraoperative complications included hypotension (n = 6) and nausea (n = 3). All of them were free from the morbid postoperative complications except wound dehiscence occurring in four patients. PMID- 12649870 TI - [Anesthetic management of a patient with oculopharyngodistal-myopathy]. AB - Oculopharyngodistal-myopathy (OPDM) is an autosomal dominant, heredofamilial myopathy accompanied with slowly progressive ptosis and extraocular palsy, and weakness of the masseter, facial, and bulbar muscles, as well as distal involvement of the limbs starting around 40 years of age or later. A 54-year-old female with OPDM underwent resection of the uterus for uterus body cancer. We speculated the patient might be at the risk of aspiration pneumonia, prolonged respiratory depression, and malignant hyperthermia, and chose spinal and epidural anesthesia. The operation was performed successfully and the patient was discharged uneventfully. PMID- 12649871 TI - [A case of re-expansion pulmonary edema after one lung ventilation for the radical operation of lung cancer]. AB - A 64-yr-old man was admitted to our hospital for the examination of the abnormal shadow in his left lung field, which was diagnosed later as lung cancer. Radical operation was scheduled under combined epidural/general anesthesia. One lung ventilation was performed to facilitate the operative procedure for two hours and fifteen minutes. About two hours after the re-expansion of his left lung, he complained of dyspnea. He was hypoxic and moist rale was audible in his left lung. Cloudy shadow emerged in the left field of his chest X-P. Under bronchofiberoptic observation, excessive serous secretions was seen, but no obstructive lesions were found in his bronchial tree. Re-expansion pulmonary edema was suspected for these findings. It was improved with mechanical ventilation with PEEP. Re-expansion pulmonary edema seldom occurs in one lung anesthesia. Although radical operation of esophageal cancer performed six years before might have induced the development of re-expansion pulmonary edema in our case, complete collapse with quick re-expansion of the lung is clearly a potential risk of re-expansion pulmonary edema. Careful management is necessary after one lung ventilation. PMID- 12649873 TI - [Difficult tracheal intubation in a patient suspected of malignant hyperthermia]. AB - We report anesthetic management of a patient suspected of malignant hyperthermia with difficult tracheal intubation. A 64-year-old man was scheduled for a fixation of humerus bone fractures in prone position. He had a history of difficult tracheal intubation due to masseter spasm, and his niece was suspected to be malignant hyperthermia. Anesthesia was induced with propofol using a target controlled infusion. No muscle relaxant was given and spontaneous breathing was maintained. Trials for tracheal intubation failed whenever using a standard laryngoscope, a bronchofiberscope, a laryngeal mask airway or an intubating laryngeal mask airway. Resecting the epiglottic elevating bar of an intubating laryngeal mask airway enabled fiberoptic tracheal intubation. No symptom suggesting malignant hyperthermia developed. PMID- 12649874 TI - [Anesthetic management for a patient with hyper-IgE syndrome]. AB - Hyper-IgE syndrome is a rare immunodeficiency disorder characterized by recurrent skin and pulmonary infections and extremely elevated serum levels of IgE. A 6 year-old girl with hyper-IgE syndrome underwent appendectomy. Anesthesia was induced with sevoflurane. After insertion of a laryngeal mask, anesthesia was maintained with nitrous oxide, sevoflurane, and oxygen. There were no perioperative complications related to anesthesia and surgery. PMID- 12649872 TI - [Anesthesia for cesarean section in a patient with transient diabetes insipidus]. AB - A 32-year-old pregnant female was admitted to our hospital at 32 week gestation and was scheduled for emergent cesarean section because of fetal distress. She had been suffering hydrodipsia and dry mouth, and had lost 4 kg in 2 weeks. Hypernatremia, hyperchloremia, and lower urinary specific gravity were preoperatively noted. Her electrolyte imbalance was partially corrected by the infusion of 1400 ml of 5% glucose solution and 500 ml of acetated Ringer's solution, but unexpected hyperglycemia; 440 mg.dl-1, appeared before surgery. Cesarean section was successfully performed with spinal anesthesia. A 1566 g male infant was delivered with 1 and 5 min Apgar scores of 2 and 1. Hyperglycemia and secondary hypoglycemia occurred in the infant in the neonatal ICU. The mother's fluid loss, including blood and amniotic fluid, was estimated at 784 ml. Five hundred milliliters of acetated Ringer's solution and 1000 ml of half saline solution with 2.5% glucose were infused before delivery, followed by the glucose solution containing a low concentration of sodium after delivery. After surgery, high serum osmotic pressure and paradoxically low urinary osmotic pressure were found. The plasma antidiuretic hormone level was normal against the high serum osmotic pressure. The electrolyte imbalance and urinary osmotic pressure were improved by using I-deamino-8-d-arginine vasopressin, and DI was finally diagnosed. Hormonal therapy was discontinued on day 20, and the patient was discharged on day 21. Some pregnancies are complicated by transient DI. Anesthesiologists have to consider DI when a pregnant female has symptoms of dehydration and a significant electrolyte imbalance. PMID- 12649875 TI - [The intubating laryngeal mask for a patient with cerebrospinal fluid rhinorrhea and pneumocephalus]. AB - A 53-year-old male was scheduled for repairs of cerebrospinal fluid (CSF) rhinorrhea and pneumocephalus under general anesthesia. He had undergone a neck clipping for a ruptured anterior communicating aneurysm 13 days before. As he had a difficult airway, Trachlight was used for successful tracheal intubation. To avoid tension pneumocephalus, an intubating laryngeal mask (ILM) was inserted with rapid sequence induction without positive pressure ventilation. A 7.0 mm ID straight silicone reinforced tube was then inserted through the ILM using a fiberscope. A fiberscope guided tracheal intubation via the ILM is recommended for patients with CSF rhinorrhea and pneumocephalus especially when the trachea is difficult to intubate under direct laryngoscopy. PMID- 12649876 TI - [Bronchospasm during upper gastrointestinal endoscopy under sedation]. AB - We experienced a case of bronchospasm during upper gastrointestinal endoscopy under sedation. An 80-year-old man came to our hospital with abdominal distension with pain, nausea and vomiting. He has the history of splenectomy, cholecystectomy for hemolytic anemia and thyroidectomy for thyroid cancer, surgery for bilateral shoulder joints and diabetes. Abdominal X-ray suggested obstruction of the small intestine. On the third hospital day, gastrointestinal endoscopy was scheduled for insertion of a long ileus tube. Under sedation with diazepam 10 mg and local anesthesia of the pharynx with lidocaine spray 24 mg, the endoscope was inserted and when it reached the esophageal-gastrojunction, respiratory rate increased to 30 breaths.min-1 with expiratory stridor. The endoscope was removed immediately. He was oro-tracheally intubated and artificially ventilated. On the fourth hospital day, he was extubated under bronchoscopy. No abnormalities were observed in the trachea, vocal cord, pharynx and larynx. Later, it was revealed that he had a history of hoarseness and dysphasia. His left recurrent nerve and cervical nerve had been resected with thyroid and right cervical nerve anastomosed to the rest of the left recurrent nerve. The insertion of upper gastrointestinal endoscope might have induced bronchospasm stimulating distal esophageal afferent vagal reflex partly by regurgitation of gastric acid under sedation. PMID- 12649877 TI - [Propofol was useful for intractable convulsion in a patient with tetanus: a case report]. AB - An 82-year-old woman with severe tetanus was admitted to our ICU. Anti-convulsive treatment was started by continuous intravenous administration of midazolam followed by thiamylal. On day 15, thiamylal became insufficient to manage the spastic convulsion without frequent intravenous administration of other sedative and muscle relaxants. On day 16, intravenous infusion of propofol was started at the rate of 1 mg.kg-1.h-1. Propofol was more effective than thiamylal to treat convulsion and to stabilize the sympathetic nerve activity. Propofol was also effective to achieve the better general condition in which enteral nutrition could be started. In this case, propofol was apparently more suitable than thiamylal to manage a patient with tetanic convulsion. PMID- 12649878 TI - [Anesthetic management for cesarian section in a patient with mitral stenosis and severe pulmonary hypertension]. AB - Mitral stenosis is one of the complicating cardiac diseases during pregnancy. We experienced cesarean section in a patient with mitral stenosis and severe pulmonary hypertension (94/41 mmHg). During surgery, complete analgesia below T 4 level was obtained by epidural anesthesia with two catheters inserted at L 3-4 and T 11-12 interspaces. By epidural anesthesia, pulmonary arterial pressure decreased to 60/30 mmHg and other hemodynamic variables were stable. A baby (BW:1880 g) was intubated because of tachypnea although Apgar scores were 6-8-8. The anesthetic course of the patient was uneventful. PMID- 12649879 TI - [Anesthetic management of a pediatric patient with a laryngeal foreign body]. AB - Laryngeal foreign bodies in children are less common than bronchial foreign bodies, but are more dangerous. We report an 8-month-old boy who accidentally inhaled an open safety pin into his larynx. Anesthesia was induced and maintained with sevoflurane, oxygen. No muscle relaxant was administered during operation. The open safety pin was removed with Jackson's esophagoscope. No complications were observed during the operation. PMID- 12649880 TI - [Anesthetic management of a hemophilia A patient with HIV infection: a case report]. AB - A 40-year-old male with hemophilia A was scheduled for right total knee arthroplasty. He was HIV positive probably due to receiving infected blood products previously. We performed the pharmacokinetic study of factor VIII in advance, which showed the increased factor VIII activity 1.6% by an injection of one unit. kg-1 of factor VIII and the half-life of about 16 hours. To keep the factor VIII activity over 100% in the perioperative period, 3000 units of recombinant factor VIII was injected one hour before the induction of anesthesia followed by continuous infusion at 125 units per hour. The factor VIII activity before the induction was 110.6%. The operation was successful and there was no sign of bleeding tendency. The factor VIII activity after the operation, however, was unexpectedly low (73.1%), and it was necessary to increase the infusion rate to 150 units per hour. The factor VIII activity was kept over 80% until POD 7 with the continuous infusion and over 60% until POD 21 with intermittent administration. Factor VIII was discontinued on POD 21 without any sign of bleeding tendency after the postoperative rehabilitation. Although this patient was HIV positive, his immune system was well controlled with HAART and there was no difficulty in the anesthetic management. To prevent accidental infection to the medical staff, we again recognized the importance of standard precautions. PMID- 12649881 TI - [Anesthetic management of a pregnant patient with congenitally corrected transposition of the great arteries for emergency cesarean delivery]. AB - A 30-year-old woman at 35 weeks' gestation with congenitally corrected transposition of the great arteries (CCTGA), whose associated anomalies were VSD, ASD and pulmonary stenosis, underwent emergency cesarean delivery under general anesthesia. She had hypoxemia due to right-to-left shunt. We discussed with obstetricians and pediatric cardiologists about her at 25 weeks' gestation. Discussed points included: 1) delivery after 30 weeks' gestation, 2) her cardiac function and anomalies, 3) cesarean delivery under general anesthesia in case of emergency, 4) management of a sleeping baby after cesarean delivery by pediatricians. Cyanosis and heart failure were noted worsening during pregnancy. General anesthesia was induced by midazolam and fentanyl following cannulation of radial artery and right internal jugular vein for blood pressure and CVP monitoring. Anesthesia was maintained with sevoflurane and fentnyl. Dopamine and dobutamine were used to support blood pressure and cardiac function. After delivery, because of decreased CVP, lactated Ringer's solution was administrated. After operation, the patient was hemodynamically stable and trachea was extubated. Anesthetic management of the patient depends on the anatomical defects of CCTGA and associated conditions. Anesthesiologist should obtain information about a pregnant woman with CCTGA from cardiologists and obstetricians in early gestation to manage patient successfully. PMID- 12649882 TI - [One-lung ventilation using a new original connector: with a bronchial blocker and a single lumen tracheal tube]. AB - We experienced 22 consecutive anaesthetic managements requiring one-lung ventilation for thoracotomy. We used our original connector, which enabled us to insert a bronchial blocker cut from Univent tube and fibreoptic bronchoscope into the single lumen tracheal tube without air leakage and instability. The bronchial blocker was easily introduced into the mainstem bronchus of the non-dependent lung. The pulmonary alveoli were well collapsed through air-vent inside the bronchial blocker, and we could offer good surgical field. When one-lung ventilation was finished during surgery, we removed the bronchial blocker with connector, and could insert a larger fibreoptic bronchoscope and a suction catheter into the single lumen tracheal tube. We could detach or exchange the bronchial blocker, when they are not functioning well. This method was excellent in the airway management for usual thoracotomy. PMID- 12649883 TI - [Ralph Milton Waters and 75 years of professionalism in anesthesiology]. AB - This short biography introduces Ralph Waters to Japanese speaking people. Meeting in Madison in June 2002, "Ralph Waters and Professionalism in Anesthesiology A Celebration of 75 Years", is also reported briefly. Ralph Waters is relatively unknown in Japan probably because during his professional life, Japan looked to the continental Europe to learn medicine and sent students chiefly to this area. After the end of the World War II, however, Japan sent a number of students to the United States including many who received the training in anesthesiology in the departments headed by the disciples of Waters or in the departments under the strong influence of Waters' philosophy. Therefore, it is probably true to say that Japanese anesthesiology today owes much to Waters' teaching. PMID- 12649884 TI - Treatment of ADHD. PMID- 12649885 TI - [On the lifestyle of the aged from the view point of holistic medicine]. AB - The population of the elderly in Japan has rapidly increasing. I want to introduce the new movement of "the new elder citizens of over the age of 75 years old". The average life expectancy of Japanese newborn is 77.10 years for males, and 84.62 for females. Yet the birth rate in Japan is low (1.22). Therefore this will further reduce the number of the productive and young age group would support the seniors. This is why the elderly support ration will come down by more than half eventually to end with 50%. In preparation for such changes to occur ageism, which is an attitude of discrimination according to age, must be vanished. Aged people should be evaluated by individual physical and mental strength, rather than by mere chronological age. Forty years ago, definition of seniors in Japan was "age 65 and over". At that time in the 1960's the average life span of the Japanese was 68 years. However, in the subsequent 40 years, the average life expectancy is in around to 81 years. So, I believe that seniors should not take care of by those from age 20 to 64; rather, they should be self reliant and engaged in social activities. In view of this, in September 2000, I proposed a change in defining seniors. In stead of "age 65 and over", seniors should be "age 75 and over". As the result, the association for the New Elder Citizens has been organized. Those of us who turn 75 in the year 2000 were 20 years when World War II ended in 1945. I believe those seniors over 75 have a mission to tell the next generation the many lessons we have learned during wartime experiences. Besides it is recommended that those new elder citizens are encouraged to live their further life under the following slogans: 1. to love 2. to begin and create 3. to preserve (with hardships, so as to learn sympathize with others who are under strain). We already started the cohort study of coming 10 years of those new elder citizens by investigating annual physical check up and their behavior and environmental elements as well as genetic study of aging process including senile dementia. Many of these Japanese elderly people have selectively good health physically, mentally and spiritually. Japanese word "Kenko" (health) consists of two Chinese characters. The first means a state of great strength, fullness and self-support. The second character of Ken-ko means "peacefulness", or serenity of mind without stress. It means holistically healthy entity. The root of the English term "Health" goes back to the 16th century Anglo Saxon word, "Hal". It embodies such concepts as "whole" or "heal" or "holy". It means that a healthy state prevails in the whole human being even if individual parts of body may be afflicted in some ways. As time passed, the human body necessarily declines in its functional capacities. However the elderly persons do have high spirituality. However he or she may continue to feel a sense of fulfillment and serenity. Those are hope that even if we have an illness or a handicap, we can still be reborn as healthy individual. Being holistically healthy means that these aspects of wellness "body", "mind" and "spirit" are integrated into the well being of a whole person. The New Elder Citizens are invited to participate the cohort study for 10 years. The results would suggest that their way of living and environmental factors would reveal most important as compared with the hereditary gene factors. PMID- 12649886 TI - The psychotherapy of psychotic patients: current status. PMID- 12649887 TI - [On the problems of diagnosis of PTSD and its legal management]. AB - Since DSM-III in 1980 and DSM-IV in 1994 were enacted, "traumatic stress" meant mainly severe bio-psycho-social stress which cause some trauma, not only psychological but physiological and social stress to the person. However, PTSD was defined as to cause some psychological stress, with criterion of A to D. The concept of PTSD are used only to psychological trauma and stress and not include physiological trauma and stress. When psychological trauma caused some psychological stress, it become a legal problem of compensation.... However, the degree of psychological trauma and stress are sometimes difficult to evaluate objectively. To prevent Malingering or Factitious Disorder, it needs to establish a strict diagnostic criteria of psychological trauma and stress, in PTSD. In particular, the definition of Trauma, which is not limited to psychological trauma should be evaluated objectively, i.e. it should be more, scientifically defined, and measured by some evaluations. PMID- 12649888 TI - Diagnosis and compensation of PTSD (post traumatic stress disorder) in Korea. PMID- 12649889 TI - Compensation for victims of trauma in the United States. AB - The awarding of damages in civil litigation is a basic element of the civil justice system in the United States. The system is not free of problems and controversy. Cases in which Posttraumatic Stress Disorder (PTSD) is the basis for a claim of damages pose specific problems in quantifying the harm done and attributing causation. This paper will provide an overview of the American civil litigation system and address some of the challenges confronted when PTSD and other emotional distress claims are made. In addition, it will discuss alternative methods for compensating victims of trauma that do not utilize the traditional civil litigation system. PMID- 12649890 TI - Problems in diagnosing and deciding on compensation for PTSD in Japan. AB - In Japan, the diagnosis of posttraumatic stress disorder (PTSD) has attracted wide attention after the Great Hanshin Earthquake and the subway sarin gas attack in 1994. Compensation problems about PTSD diagnosis has became a social problem now. So we felt the necessity to create a clear criteria to be used by Japanese psychiatrists for diagnosis PTSD. We send out a questionnaire about diagnosis and compensation for PTSD. We received back 764 questionnaire which equaled 21.7% of total that we send out. In the questionnaire we set up fictitious examples based on real cases, and asked the psychiatrists who participated in the survey whether these cases could cause PTSD. 1 explain the criteria used by Japanese psychiatrists for diagnosis PTSD and also to report PTSD examples which became to be social problem. PMID- 12649891 TI - [Civil and criminal suits concerning PTSD in Japan]. AB - In civil and criminal trials in Japan, post-traumatic stress disorder (PTSD) issues have recently become a common topic of debate. In both types of (civil and criminal cases) judgments have been either affective or negative to PTSD. Contributing factors to the diagnosis of the accused that is identified to suffer from PTSD vary between cases. The difference in the establishment of the diagnosis for PTSD between psychiatrists reflects an aspect of the current situation in Japan where they have not used perfectly obvious and clearly defined diagnostic criterion, including ICD-10 and DSM-IV. To resolve this problem, a simple, clear and objective criterion of PTSD must be established. Instead of criterion that may take a long time to be established, a checklist can be used to relate PTSD with legal matters, as well as to determine liability in damages of the accused in civil cases and punishment in criminal cases. PMID- 12649892 TI - [Recent trends in work-related compensation involving job-related suicide and mental disease]. AB - According to National Police Agency statistics, suicide and the number of work related accident claims involving mental disease is rapidly increasing in Japan. In addition, the number of lawsuits against companies and government employers involving mental health issues has substantially risen. The boundary between job related mental disease and non-job-related mental disease is becoming increasingly ambiguous. Today, I am going to highlight problems in the diagnosis of job-related mental disease. It is very important to educate leaders in the business community as well as laborers about the prevention of job-related mental disease, especially regarding suicide and its warning signs. I am going to give examples of some lawsuits and highlight some deficits in the present guidelines for diagnosing mental disease in work-related compensation claims. As it is predicted that the prevalence of mental disease will continue to increase, the criteria for diagnosing mental disease and the terms and methods of medical treatment in regards to compensation should be reviewed. PMID- 12649893 TI - [Psychopathological features of "primary social withdrawal"]. AB - This article attempts to clarify the psychopathological features of "primary social withdrawal". Since the concept of "social withdrawal" in general may include various diseases, we isolated and defined "primary social withdrawal" by excluding cases that could be diagnosed by established classifications of mental disorders. First, we examined two cases of "primary social withdrawal" which could not be classified using DSM-IV. We then compared these patients' psychopathological features to those seen in apathy syndrome, taijin kyofu sho and personality disorders. Based on this comparison, we identified five pathological features of "primary social withdrawal": (1) episodes of defeat without a struggle, (2) protecting the ideal image of the expected self, (3) parents' investment in that ideal self, (4) holding an ideal self image shaped more by the desires of others than those of themselves, and (5) avoidance behavior to maintain the positive opinion of others. Finally, we discuss the socio-cultural background of social withdrawal and possible therapies. PMID- 12649894 TI - [Competitiveness in sports activities of the psychiatrically disabled]. AB - In 2002, psychiatrically disabled athletes joined an historic first open game of volleyball at the national sports games for the disabled. Compared to the competitive sports and Paralympic Games that physically and intellectually disabled athletes have participated in, activities for the psychiatrically disabled have not been well-organized. In this paper, we examine a number of problems that have arisen when the psychiatrically disabled joined competitive sports games. We identify two major characteristics of the psychiatrically disabled of particular relevance when organizing competitive sports activities. First, all psychiatrically disabled athletes need treatment of their individual diseases. For example, psychiatric symptoms fluctuate markedly over time, unlike physical or intellectual disabilities, whose symptoms are much more stable. Exacerbations of psychiatric illness are also likely to occur due to the stresses of competitiveness. Second, psychiatric disabilities are manifestations of disorders in the central nervous system, which makes the classification of psychiatric disabilities less straightforward than classification of the physically disabled. These two characteristics require special attention when organizing competitive athletic challenges that include the psychiatrically disabled. However, promoting sports activities that include the psychiatrically disabled would be expected to reduce the prejudice toward and subsequent social disadvantages experienced by psychiatric patients. Thus, with careful planning to successfully integrate psychiatrically disabled athletes, we expect increased promotion of such sports activities in the future. PMID- 12649895 TI - [Molecular mechanisms of biological clock: from molecular rhythms to physiological rhythms]. PMID- 12649896 TI - [Photoreception and circadian clock system in the pineal gland]. PMID- 12649898 TI - [Sleep-wake rhythm disorders]. PMID- 12649897 TI - [Temporal anticipatory behavior and circadian clock]. PMID- 12649899 TI - [Sleep substances]. PMID- 12649900 TI - [SPECT imaging using [123I]beta-CIT and [123I]IBF in extrapyramidal diseases]. AB - Imaging of dopaminergic function is useful in the investigation of patients with Parkinson disease (iPD) and other extrapyramidal diseases. Using agents that bind to dopamine transporters ([123I]beta-CIT) and receptors ([123I]IBF SPECT), we investigated SPECT in 9 healthy volunteers and 24 patients for dopamine transporters as well as 15 patients for dopamine receptors. In beta-CIT SPECT studies, we examined 17 iPD patients (63.3 +/- 9.9 y/o), 3 multiple system atrophy (MSA) patients (OPCA type) (64.0 +/- 8.0 y/o), 2 vascular parkinsonism (VP) patients (71.0 +/- 0.0 y/o), 1 progressive supranuclear palsy (PSP) patient (69 y/o), 1 cortico-basal degeneration (CBD) patient (50 y/o) and nine healthy controls (39.1 +/- 9.3 y/o). For IBF SPECT studies 11 iPD patients (60.6 +/- 10.9 y/o), 3 MSA patients (2 OPCA type (50.5 +/- 3.5 y/o) and 1 SND type (65 y/o)) and 1 PSP patient (60 y/o) underwent SPECT scans after the injection of [123I]IBF. The specific to nonspecific striatal uptake ratio(St/Oc-1), ratio of putaminal uptake to caudatal uptake (Pu/Ca), and asymmetry indices (AI) were estimated. beta-CIT studies showed St/Oc-1 as follows; iPD: 2.66 +/- 1.09 (n = 17), VP: 5.73 and 7.39, MSA: 1.84 +/- 0.46 (n = 3), PSP: 2.34, CBD: 2.16. In all extrapyramidal diseases except VP, St/Oc-1 ratios were significantly lower than those in normal volunteers (6.46 +/- 1.08) (p < 0.01). Also in early-phase iPD patients (Yahr I II), St/Oc-1 (3.16 +/- 1.49: n = 4) was significantly lower than those in normal volunteers (p < 0.01). In IBF studies, St/Oc-1 ratios were significantly higher in early-phase (Yahr I-II) iPD patients (1.82 +/- 0.25: n = 5) than those in late phase (Yahr III-IV) iPD patients (1.38 +/- 0.32: n = 6) (p < 0.05). The Pu/Ca ratios in iPD patients (1.12 +/- 0.13) and MSA (OPCA type) patients (0.95 +/- 0.05) were higher than that in MSA (SND type) patient (0.78) and were lower than that in PSP patient (1.55). In conclusion, beta-CIT-SPECT is useful for the diagnosis of early-phase iPD patients and for differentiating VP from other extrapyramidal diseases. IBF-SPECT is useful for the diagnosis of the severity of iPD and has the possibility for ruling out MSA (SND type) or PSP from iPD. Both tracers are useful for investigating the pathophysiology of patients with iPD and other extrapyramidal diseases. PMID- 12649901 TI - [A case report of tuberculous meningitis: limits of detection using polymerase chain reaction (PCR) and treatments]. AB - A 40-year-old man with intractable meningitis was transferred to our hospital 6 weeks after onset. On admission, he showed consciousness disturbance, meningeal signs and right oculomotor nerve palsy. MRI demonstrated prominent cisternal enhancement and hydrocephalus. We suspected tuberculous meningitis as the diagnosis, and treated with antituberculotics, though he died of midbrain infarction day 11 of the treatment. Before and during the admission, bacterial cultures, PCR, smear examination of cerebrospinal fluid(CSF) were repeated. But no evidence of tuberculosis was obtained. Cultures of the brainstem fragments detected mycobacterium, which was finally confirmed by a PCR method after his death. PCR provides a rapid and reliable diagnosis of tuberculous meningitis, although there is a potential for false-negative. Thus the clinical, radiological and CSF findings should be stressed. Corticosteroids treatment should be considered in cases with ischemic lesions. PMID- 12649902 TI - [A case of cerebral deep venous and venous sinus thromboses with anti-cardiolipin antibody]. AB - We describe a case of cerebral deep venous and venous sinus thromboses with anti cardiolipin antibody. A 62-year-old male with no previous illness of thrombosis but with alcohol abuse was admitted with acute onset unconsciousness. He recovered two days after with no severe sequela. Laboratory findings suggested the preceding conditions of dehydration and inflammation. X-ray CT of the head revealed symmetrical low density areas in the thalami and basal ganglia, high density signs in the cerebral deep veins, and dilation of the lateral ventricles. MRI on the second hospital day showed abnormal intensities in the thalami and basal ganglia (high signal on T 2-weighed and FLAIR image, low signals on T 1 weighed image, but almost isointensity on diffusion weighed image) and acute to subacute phase thrombus in the superior sagittal sinus. Abnormal intensities observed on MRI disappeared gradually in the following studies. Venous phase images of cerebral angiography performed in chronic phase disclosed occlusion of the superior sagittal sinus and stenosis of the vein of Galen. These radiological findings support the diagnosis of cerebral deep vein and venous sinus thromboses. Hematological examination revealed positive anti-cardiolipin IgG antibody. Several cases of cerebral deep venous thrombosis with anti-cardiolipin antibody have been reported. In our case, dehydration induced by alcohol abuse would have been the trigger of thrombosis, while the existence of anti-cardiolipin antibody might contribute to the risk of thrombosis as an underlying condition. PMID- 12649903 TI - [Hypertensive encephalopathy detected by MRI]. PMID- 12649904 TI - [Claude's syndrome caused by midbrain hemorrhage]. PMID- 12649905 TI - [A 65-year-old woman with progressive loss of vision and visual field defects]. AB - We report a 65-year-old Japanese lady who suffered from progressive loss of vision and visual field defect. She was well until her 61 years of the age in November of 1999, when she was found to have bitemporal hemianopsia. A small enhancing mass lesion was found in the chiasmatic region. She was treated with steroid and she noted marked improvement in her visual field defects. In August of 2000, she noted disturbance of gait. Cranial MRI revealed a mass in the right midbrain extending into the hypothalamic and thalamic regions. She was again treated with steroid with marked improvement. However, in November of 2001, she started to show somnolence and diabetes insipidus. She was treated with steroid, nasal desmopressin, and insulin for her steroid induced diabetes mellitus. Cranial CT scan showed a large enhancing lesion involving the entire midbrain, hypothalamus, and the thalamic regions. She developed respiratory arrest on July 15, 2001 and was pronounced dead. She was discussed in a neurological CPC and the chief discussant arrived at the conclusion that the patient had a primary malignant lymphoma of the brain. Clinical diagnosis in the early stage of her disease was neurosarcoidosis. Post-mortem examination revealed a mass continuously involving the pons, midbrain, hypothalamus, thalamus, and the putamen. The optic chiasm was enlarged. By histologic examination, the mass consisted of dense medium sized tumor cells. Immunohistologic observation revealed that the tumor cells were B-cell type malignant lymphoma. No tumor cells were found in the systemic organs. PMID- 12649906 TI - [Comparative study of hypothermic circulatory arrest and normothermic distal perfusion for thoracoabdominal aortic aneurysm repair]. AB - BACKGROUND: Hypothermic bypass with circulatory arrest for thoracoabdominal aortic aneurysm (TAAA) repair is employed for its protective effect on spinal cord function and because it avoids clamping the diseased aorta. However, organ dysfunction caused by reperfusion injury as well as bleeding tendencies due to deep hypothermia have been described. In this paper we compared the efficacies of the hypothermic and normothermic operations. METHODS: Between February 1996 and June 2000, 28 patients underwent thoracoabdominal aortic repair. Twenty-three patients were men, 5 were women, and the median age was 55.3 (range 23 to 75 years). Fourteen patients had aortic dissection, and 7 had Marfan syndrome. Fourteen patients required reconstruction of visceral arteries. Twelve patients underwent TAAA repair under deep hypothermic circulatory arrest (H group), and 15 under normothermic distal perfusion (N group), while 1 patient underwent a simple clamp procedure. Perioperative data and early outcomes were compared between groups. RESULTS: The overall 30-day mortality rate was 0%, but 3 patients (25.0%) in II group, and 1 patient (6.3%) in N group died during hospital stay. Operation time and bypass time were longer in H group than N group (operative time 793 min vs. 481 min (p < 0.05): cardiopulmonary bypass (CPB) time 255 min vs. 102 min (p < 0.05). Also, more intraoperative bleeding was found in H group than in N group (3,506 ml vs. 1,220 ml). Spinal cord neurologic deficit did not occur in either group. Respiratory failure occurred in 3 patients (25.0%) in H group and one (6.3%) in N group. Renal failure occurred in 3 (25.0%) in H group, and none in N group. CONCLUSION: Early and mid-term outcome of TAAA repair was almost satisfactory and without neurospiral complications. The deep hypothermic operation is more likely to induce postoperative respiratory and renal dysfunction than the normothermic operation. TAAA repair using deep hypothermic circulatory arrest should be limited to patients with TAAA involving the distal arch or a severely calcified aortic wall. PMID- 12649907 TI - [Local advanced lung cancer invaded thoracic vertebral bodies with unruptured cerebral aneurysm]. AB - A 57-year-old man presented with the chief complaint of left shoulder pain in June 2001, and paridrosis of left upper trunk and left upper limb in July 2001. Head magnetic resonance imaging (MRI) showed 8 mm sized unrupture aneurysm of left middle cerebral artery, and chest computed tomography (CT) showed the lung tumor invaded thoracic vertebral bodies. The local advanced lung carcinoma (cT4N0M0) and unrupture aneurysm of left middle cerebral artery was diagnosed. The prevented clipping of unrupture aneurysm was performed at 11th September 2001, and left upper lobectomy, hemivertebrectomy and reconstruction of thoracic vertebral body (Th 3-5) with Modul' ICS at 12th October 2001. The pathological findings revealed squamous cell carcinoma. The staging was pT4N0M0, IIIB. The postoperative course was uneventful. After the radiotherapy (50 Gy), chemotherapy (gemcitabine and vinorelbine) was performed. But the radiation pneumonia was occurred and chemotherapy was intermitted. The steroid was administrated due to the radiation pneumonia, and the complication was improved. He discharged at 17th April 2002 and had no recurrence. The prevented clipping of unrupture cerebral aneurysm and the reconstruction of thoracic vertebral body (Th 3-5) with Modul' ICS were useful for the radical operation of the local advanced lung cancer. PMID- 12649908 TI - [Resection of freely mobile atheroma in the ascending aorta under circulatory arrest during coronary artery bypass grafting surgery]. AB - A 52-year-old male with a 13 years history of hemodialysis developed unstable angina. Preoperative examination revealed critical stenoses in 3 coronary arteries and extensive calcification in the ascending aorta. During urgent coronary artery bypass surgery, epiaortic ultrasonography demonstrated a large and markedly mobile atheroma in the ascending aorta. Therefore, he underwent resection of this atheroma using cardiopulmonary bypass and circulatory arrest. His postoperative course was fine. This aggressive strategy for a diseased aorta can be a viable option in selected cases. Epiaortic ultrasonography appeared to be indispensable during surgery for patients like a present one. PMID- 12649909 TI - [Postoperative recurrence and medical economics of video-assisted thoracic surgery (VATS) for spontaneous pneumothorax]. AB - We analyzed 96 patients undergoing video-assisted thoracic surgery (VATS) for pneumothorax of our institute (Otsu Red Cross Hospital) to determine the recurrence rate and to analyse various factors relating to recurrence and cost performance. From January 1997 to November 2000, we encountered 96 cases (78 male, 18 female, 14-72-year-old mean age 31.1). In 6 cases, pneumothorax had recurred (recurrent rate 6.3%), including 5 mals and 1 female (mean age 28.0). The relationship between recurrence and various factors including age, preoperative procedure, radiological bulla diagnosis, operation time, number of bulla or blebs, location of bulla, intraoperative treatment, post-operative air leakage, duration of postoperative drainage and duration of hospitalization was calculated. Preoperative diagnosis of bulla on chest X-ray film and computed tomography (CT) was significantly calculated a higher recurrence rate after VATS operation. Cost, age and operation time were significantly correlated, r = 0.514, r = 0.436, respectively, but length of hospitalization was not significant. To analyze cost performance, we investigated the correlation between cost/day and various factors. A negative correlation was noted between cost/day and the length of hospitalization (r = -0.739). In summary, recurrence rate of postoperative pneummothorax was 6.3% in our institute. When bulla was radiologically diagnosed preoperatively, recurrence tended to occur postoperatively. The negative correlation between cost/day and length of hospital stay suggests that the length of hospital stay should be reduced to as short as possible in terms of cost reduction. PMID- 12649910 TI - [Usefulness of mini loop retractor in video-assisted thoracic surgery]. AB - We have developed a technique using a Mini Loop Retractor II and successfully performed video-assisted thoracoscopic surgery (VATS) in 32 patients. It is inserted into the pleural cavity through a skin puncture and a loop at its distal end is easily, and is freely adjustable in dimensions so that it can act as a retractor or holding forceps. Mini Loop Retractor II was very useful instruments in VATS. PMID- 12649911 TI - [Semi-automatic suturing device for minimally invasive cardiac surgery]. AB - Semi-automatic suturing device for minimally invasive cardiac surgery (MICS) was developed. Exchange of the needle is attained by an automatic grasping action and the needle shift parallel to the long axis of the device. This device (5 mm in diameter) can be inserted through a 5 mm trocar. This new device facilitates not only operative procedure such as mitral valve replacement and patch closure during MICS but conventional cardiac surgery. PMID- 12649912 TI - [Infected false aneurysm due to methicillin-resistant staphylococcus aureus after proximal aortic grafting]. AB - A 70-year-old man developed meticillin-resistant staphylococcus aureus (MRSA) mediastinitis after prosthetic graft replacement of the ascending aorta. The sternal wound was reexplored and a single-stage procedure of irrigation, debridement, and omental transposition was performed. Ten months after the first operation, he suffered recurrence of pyrexia and the presence of false aneurysm originated from the distal suture line was diagnosed by the chest computed tomography (CT) scan. Re-replacement of the ascending aorta and proximal hemiarch with rifampicin soaked Gelseal was successfully performed. Hypothermic perfusion with circulatory arrest through peripheral cannulation and left ventricular venting via a left anterior thoracotomy was useful to obtain safe reentry in the operation of retrosternal false aneurysm. PMID- 12649913 TI - [Re-dissection of aortic root without gelatin-resorcin-formalin (GRF) glue effect one year after aortic repair]. AB - A 58-year-old man, who had undergone emergency graft replacement of ascending and total aortic arch using gelatin-resorcin-formalin (GRF) glue 1 year before, complained of progressive shortness of breath. Ultrasound cardiography revealed severe aortic regurgitation and dissecting aneurysm of aortic root. He underwent composite graft replacement with the aid of the right axillary artery perfusion and deep hypothermic circulatory arrest. The redissected brownish intima was identified in the area of noncoronary cusp. There was no special finding in the subsequent histopathological examination. The use of GRF glue for reconstructing the dissected aortic root is associated with a certain amount of risk of aortic wall redissection. Therefore, care should be taken to ensure proper use of GRF glue. PMID- 12649914 TI - [Traumatic aortic regurgitation with an ascending aortic aneurysm: report of a case]. AB - We reported a case of an aortic regurgitation due to nonpenetrating chest trauma with an ascending aortic aneurysm. A 58-year-old man was admitted to our hospital with an acute left cardiac failure. Severe aortic regurgitation and ascending aortic aneurysm were diagnosed on ultrasonic echocardiography (UCG) and computed tomography (CT) scan. On the 13th day after the injury, semi-emergency operation was done. The commissure between the right coronary and non-coronary cups was torn, and modified Bentall operation was performed. The patient recovered well and discharged uneventfully. PMID- 12649915 TI - [Ventricular pseudo-false aneurysm after myocardial infarction; report of a case]. AB - A 72-year-old man was admitted to our hospital with chest discomfort and syncope. The echocardiography showed the left ventricular aneurysm. The coronary arteriography revealed a complete occlusion at distal portion of the right coronary artery and anterior aneurysm of the left ventricle was demonstrated. We diagnosed that it was false aneurysm because of communicating with the left ventricle through a small orifice. Surgical repair was carried out after resection of the aneurysm of the left ventricle. The postoperative course was uneventful and discharged on the 33rd day after surgery. Pseudo-false ventricular aneurysm of the left ventricle was diagnosed by histological examination. PMID- 12649917 TI - [Coronary subclavian steal syndrome; report of a case]. AB - A 64-year-old man was admitted to our hospital with chief complaint of chest discomfort. He received coronary artery bypass grafting utilizing the in situ left internal thoracic artery 10 years ago. Coronary and left subclavian artery angiogram revealed coronary subclavian steal syndrome and 90% stenosis in the proximal left subclavin artery. Ultrasonography of neck vessels demonstrated 75% stenosis in the bifurcation of left carotid artery. We performed axilloaxillary artery bypass grafting to avoid brain ischemia. Myocardial thallium scintigraphy on dipyridamole testing after axilloaxillary artery bypass grafting could not detect myocardial ischemia. Axilloaxillary artery bypass grafting was effective for coronary subclavian steal syndrome. PMID- 12649916 TI - [The isolated congenital mitral regurgitation due to a partial defect of the anterior leaflet]. AB - The isolated congenital mitral regurgitation (MR) is a rare anomaly. We have successfully treated a 7-year-old boy who had MR due to a partial defect of the anterior leaflet. Echocardiogram revealed moderate MR at the anterior leaflet as a result of its partial defect. Another MR was also detected at the coaptation zone of the 2 leaflets due to mild prolapse of the anterior leaflet. The mitral valve was approached via atrial septostomy and a partial defect (8 x 6 mm) was found near the postero-medial commissure of the anterior leaflet. Any chordae did not develop around the defect or subvalvular lesions. Other parts of the leaflet was seemed normal and any inflammatory or degenerative changes were not detected in the rest of the leaflets. We have done direct closure of the defect of the leaflet and also added annuloplasty at the posteromedial commissure by modified Kay method. PMID- 12649918 TI - [Combined operation for left ventricular aneurysm and ventricular septal perforation with normal coronary arteries; report of a case]. AB - We experienced a rare case of left ventricular aneurysm (LVA) and ventricular septal perforation (VSP) with normal coronary arteries, and had a good outcome by undergoing Dor procedure and direct closure of VSP individually. A 68-year-old female was printed out cardiac systolic murmur. Echocardiogram revealed LVA and VSP, but coronary arteriogram showed no significant stenosis. The apex was incised longitudinally, lateral and parallel to the interventricular groove. VSP was directly closed by 2 pieces of 3-0 polypropylene suture with felt strip. Purse-string stitch with 3-0 polyester suture around the transitional zone between fibrotic tissue and viable tissue was secured to firm fibrous tissue. The residual orifice was closed with an elliptical composite patch of equine pericardial patch and polyester velour with 3-0 polypropylene suture. We considered based on the operative finding that myocardial infarction (MI) led to LVA and VSP. Coronary thrombosis rather than coronary spasm may have provoked MI because spasm provocative test was negative. PMID- 12649919 TI - [Successful transection of racemose hemangioma of bronchial artery; report of a case]. AB - A 52-year-old female was admitted to the hospital because of increasing hemoptysis for twenty years. Chest computed tomography (CT) on admission showed bronchoectasia with dilated bronchial arteries in the left lower lung field. Bronchial arteriogram showed racemose hemangioma of bronchial arteries draining to left lower pulmonary arteries. Transresection of bronchial artery and partial lung resection was performed. The cut surface of the resected specimen showed bronchoectasia with dilated bronchial arteries (8 mm in diameter). She is well 15 months following the treatment with no evidence of hemoptysis. PMID- 12649920 TI - [Mediastinal mature teratoma perforating into the lung with elevated serum carbohydrate antigen 19-9 (CA19-9) levels; report of a case]. AB - A 17-year-old woman was suddenly seized with anterior chest pain and admitted to our hospital. Chest X-ray, computed tomography (CT) scan and magnetic resonance imaging (MRI) showed a mass shadow in the left middle lung field associated with the left pleural effusion and high serum carbohydrate antigen 19-9 (CA 19-9) levels (58.5 U/ml). We performed a total resection of the tumor with adherent parts of the left lung. The tumor was 6.5 x 3. 8 x 2.9 cm in size, and was made up of soft tissues, fluid and cystic components. The histological diagnosis was a mature cystic teratoma with perforation into the lung, which contained pancreatic tissues, hepatic cells, bronchial epithelium, serous glands and so on. The levels of fluid amylase and CA 19-9 were high. We report mediastinal mature teratoma perforating into the lung and discuss the mechanism of perforation. PMID- 12649921 TI - [Torsion of the middle lobe after right upper lobectomy of the lung; report of a case and the review of the Japanese literatures]. AB - We report a case of torsion of the residual right middle lobe of the lung, following right upper lobectomy for lung cancer. A 71-year-old man who had medical treatment for emphysema was admitted with a lung tumor on chest computed tomography. The tumor was diagnosed as pulmonary adenocarcinoma by transbronchial biopsy. Right upper lobectomy with mediastinal lymph node dissection, and partial resection of the right lower lobe were performed. On the following day, chest X ray showed an opacification in the right upper lung field, which gradually increased. Bronchoscopic examination revealed a stenotic middle lobe bronchus. Torsion of the middle lobe was suspected, and rethoracotomy was performed on the second postoperative day. The middle lobe was torsed 90-degree counterclockwise around its bronchovascular pedicle. A middle lobectomy was performed secondary to severe congestion. The patient was discharged in good condition on the 11th postoperative day. In reviewing the literatures including this case, 13 of 16 torsions occurred after right upper lobectomy of the lung. Thirteen patients had rethoracotomy, 10 of them underwent resection of the rotated lung. Simple detorsion was carried out in 3 patients, and 1 of them developed cerebral infarction. Lung torsion was reported to be potentially life-threatening. Therefore, fixation of a remaining lobe should be performed. Exploratory thoracotomy should be performed without delay, if lung torsion is suspected. PMID- 12649922 TI - [Mediastinal lymphangioma treated under video-assisted thoracic surgery]. AB - Mediastinal lymphangioma is a rare benign tumor that is usually seen in anterior mediastimun of the adult. A 15-year-old girl with anterior mediastinal tumor, 10 cm in length, was suspected to have lymphangioma based on magnetic resonance imaging. She underwent tumor resection under video-assisted thoracic surgery. Pathological examination reveled that tumor was composed of lymphatic ducts and smooth muscles, and diagnosis of lymphangioma was established. Both the onset of young age and location of left side of the mediastinum are considered to be unusual. PMID- 12649923 TI - [Usefulness of rapid quantitative cardiac troponin T and myoglobin assays for the diagnosis of acute myocardial infarction]. AB - BACKGROUND AND OBJECTIVES: The rapid cardiac troponin T (cTnT) test is widely used to detect myocardial necrosis in the emergency setting. This assay system is rapid and myocardial-specific, but the plasma cTnT concentration is difficult to determine quantitatively. A recently developed bedside cTnT and myoglobin (Mb) analyzer (CARDIAC system) was evaluated. METHODS: The new CARDIAC system was used to measure plasma cTnT and Mb levels, and serum levels of creatine kinase MB isoenzyme (CK-MB), cTnT and Mb were measured by conventional assays in 160 consecutive emergency patients with suspected acute myocardial infarction. RESULTS: The sensitivity of cTnT for identifying acute myocardial infarction was 76%, significantly higher than that of Mb (67%, p < 0.01) and CK-MB (54%, p < 0.05). The diagnostic sensitivities in patients admitted < or = 3 hr and 3-6 hr after onset were 52% and 65% for cTnT, 60% and 90% for Mb, and 36% and 50% for CK MB, respectively. These sensitivities of Mb were significantly higher than those of CK-MB but not cTnT. However, the sensitivity of cTnT (100%) was significantly higher than that of Mb (58%, p < 0.01) and CK-MB (70%, p < 0.001) in patients admitted > 6 hr after onset. The specificities of cTnT, Mb and CK-MB were 96%, 76% (p < 0.001 vs cTnT and CK-MB) and 95%, respectively. Therefore, cTnT (86%) had significantly (p < 0.001) higher diagnostic accuracy compared with Mb (71%) and CK-MB (75%). Combination diagnosis using cTnT and Mb showed the highest sensitivity (86%) compared with cTnT (p < 0.05) and Mb (p < 0.001). The correlation coefficients between the levels measured by CARDIAC system and those by ordinary assays were 0.98 in cTnT and 0.97 in Mb. CONCLUSIONS: Bedside rapid quantitative assays of cTnT and Mb are useful as a point of care test for the diagnosis of acute myocardial infarction. PMID- 12649924 TI - [Usefulness of coronary flow velocity measurement by transthoracic Doppler echocardiography in patients with acute coronary syndrome]. AB - OBJECTIVES: To evaluate the usefulness of left anterior descending coronary artery (LAD) flow measured by transthoracic Doppler echocardiography (TTDE) in patients with acute coronary syndrome. METHODS: Thirty consecutive patients with acute coronary syndrome in the LAD territory and unstable angina or non-ST segment elevation myocardial infarction required decisions on the need for emergency coronary angiography. The diastolic peak flow velocity was measured in the distal segment of the LAD under guidance of color Doppler echocardiography in the emergency room. If LAD flow was not detected within 10 min, the coronary flow was judged as under the detection limit. The results of TTDE were compared with the Thrombolysis in Myocardial Infarction (TIMI) grade of LAD determined by coronary angiography, which was performed within 1 week (mean 2.5 +/- 1.5 days) in all patients. RESULTS: Coronary flow was not detected by TTDE in six patients who had TIMI grade 1 or 0. The diastolic peak flow velocity in 19 patients with TIMI 3 was higher than that in 5 patients with TIMI 2 (20.1 +/- 4.1 vs 10.9 +/- 2.3 cm/sec, p = 0.0001). A diastolic peak flow velocity of 14 cm/sec was the optimal cut-off value for the prediction of TIMI 3, with a sensitivity of 95% and a specificity of 100%. CONCLUSIONS: Coronary flow velocity measured by TTDE closely reflected the TIMI grade. Coronary flow measurement by TTDE is useful to decide the treatment strategy for patients with acute coronary syndrome in the emergency room. PMID- 12649925 TI - [Long-term efficacy of combination therapy using antiarrhythmic agents and angiotensin converting enzyme inhibitor in patients with paroxysmal and persistent atrial fibrillation: importance of the timing of administration]. AB - OBJECTIVES AND METHODS: This study examined the long-term efficacy of combination therapy using antiarrhythmic agents and angiotensin converting enzyme inhibitor (ACE-I) to maintain sinus rhythm in patients with paroxysmal and persistent atrial fibrillation (Paf). There were 246 patients (176 men, 70 women, mean age 67.3 +/- 11.7 years, mean follow-up period 48.9 +/- 29.3 months) divided into two groups: the ACE-I(+) group (n = 74) and the ACE-I(-) group (n = 172). RESULTS: The incidence of hypertension and underlying heart disease in the ACE-I(+) group (85.1% and 34.3%, respectively) was significantly higher than those in the ACE-I( ) group (37.8% and 25.0%, respectively) (both p < 0.01). Left ventricular ejection fraction in the ACE-I(+) group (65.6 +/- 12.5%) was significantly lower than that in the ACE-I(-) group (71.9 +/- 8.9%) (p < 0.01). The actuarial rate of the maintenance of sinus rhythm at 48 months in the ACE-I(+) group (86.5%) was similar to that in the ACE-I(-) group (83.1%). Among the 104 patients who had suffered from Paf for < 3 months after the first episode, the actuarial rate of maintenance of sinus rhythm at 48 months in the ACE-I(+) group (97.1%, n = 35) was significantly higher than that in the ACE-I(-) group (82.6%, n = 65), and the period of maintenance of sinus rhythm in the ACE-I(+) group (54.8 +/- 30.8 months) was significantly longer than that in the ACE-I(-) group (28.4 +/- 20.5 months) (both p < 0.05). CONCLUSIONS: ACE-I must be additionally administered within 3 months of the first Paf episode to maintain normal sinus rhythm in patients with Paf. PMID- 12649926 TI - [Assessment of the clinical feasibility of OptiGo for hand-held echocardiography]. AB - OBJECTIVES: Hand-held ultrasound devices are becoming available for clinical examination, but the accuracy and precision of such devices are unclear. This study compared the accuracy of a hand-held echo device to a standard echo system. METHODS: Twenty-two patients were examined with the OptiGo (Phillips Medial Systems) hand-held ultrasound system with a 2.5 MHz transducer and SONOS 5500 (Phillips Medial Systems) standard ultrasound system with a 2 to 4 MHz wideband transducer. Patients with cardiac arrhythmia and tachycardia were excluded. Image quality, chamber size (left ventricle, left atrium), global and regional left ventricular function, valve morphology and severity of valve regurgitation were assessed. RESULTS: There was good agreement between the two imaging devices for image quality (77.3%), left ventricular ejection fraction (90.5%), regional wall motion score (> 90%), valve morphology (> 90%), severity of valve regurgitation (> 81.0%) and there was good correlation and agreement for left ventricular and left atrial size. CONCLUSIONS: Although the OptiGo has limitations, the hand-held examination appropriately estimates global and regional left ventricular function, valve morphology, valve regurgitation and chamber size. PMID- 12649927 TI - [Post-operative cardiac rehabilitation programs for early returning to daily life activities: self-measurement of heart rate and blood pressure during staying at home trial]. AB - BACKGROUND AND OBJECTIVES: Recently, early mobilization and discharge after cardiac surgery have been recommended. However, many patients are anxious about returning to daily life soon after undergoing heart operations. To resolve this problem, an individualized rehabilitation plan for each patient is important. Rehabilitation programs must estimate the level of cardiac function in daily life. This study evaluated self-measurements of heart rate and blood pressure during home-based exercise training. METHODS: Thirty-six patients, 28 men and 8 women (mean age 58 +/- 19 years) who underwent cardiac operations were enrolled in this study. None of the patients experienced postoperative complications. Changes in heart rate and blood pressure during daily activities at home were measured by the patients. This data was then used to plan individual rehabilitation programs. RESULTS: The blood pressure rose from 114 +/- 17 to 139 +/- 21 mmHg (mean increase of 25 +/- 15 mmHg) when the patients were asked to walk up and down a set of stairs. Thirteen patients (36%) exhibited an increase in blood pressure of 30 mmHg or more while ascending the stairs. The patients' blood pressure returned to its pre-exercise level after 5 min. The heart rate rose from 84 +/- 15 to 113 +/- 14 beats/min (mean increase of 29 +/- 8 beats/min) during the exercise. During the home-based training period, the maximum blood pressure was 133 +/- 22 mmHg, and the maximum heart rate was 97 +/- 13 beats/min. CONCLUSIONS: The patients were very careful during their trial outpatient period, as this was their first post-cardiac surgery experience. Consequently, the degree of exercise at home was even more mild than in hospital. Self-measurement of heart rate and blood pressure was feasible. By referring to these measurements, the patients were able to monitor and increase their level of exercise. This post cardiac surgery rehabilitation program is helpful for early returning to daily life activities. PMID- 12649928 TI - [Complex myxoma detected by syncope: a case report]. AB - A 23-year-old man was admitted to our hospital for evaluation of syncope and intracardiac masses. Echocardiography revealed three masses in the right ventricle and one in the left ventricle. The largest mass, 4 by 5 cm, occupied the right ventricular outflow tract and prolapsed through the pulmonary valve orifice. Right ventricular systolic pressure was 65 mmHg. A soft mass, 4 by 5 cm, was found on the left subcostal abdominal wall and multiple pigmented spots on the face and trunk. Histological examination of the resected tumors, including the abdominal soft mass, were consistent with myxoma. The combination of multiple cardiac and abdominal wall myxomas and pigmented skin lesions in this young patient is considered to be a diagnostic feature of Carney complex. PMID- 12649929 TI - [Cardiovascular imaging in-a-month. A 64-year-old woman showing ST depression and widening QRS during anginal attack. Unstable angina due to left main coronary artery spasm]. PMID- 12649930 TI - [Normality of bladder function. A study of young healthy volunteers]. AB - Through urodynamic evaluations of sensation and of motor function in 39 young healthy volunteers, we were able to gain information on what can be considered "normal" in lower urinary tract function and to evaluate technical urodynamic tests. Most urodynamic parameters show large variations. Numeric data considered pathological in literature are found in our symptomfree study group. The principle of urodynamic testing should be to reproduce the symptoms patients complain about during the test and then correlate these symptoms with the urodynamic data. Different urodynamic data in a normal population are presented. Several of these were not available before. PMID- 12649931 TI - Thyroid and infertility. AB - Infertility is defined as the inability to conceive after one year of regular intercourse without contraception. The prevalence of infertility is estimated between 12 and 14% and remains stable in recent years. It thus represents a common condition, with important medical, economic and psychological implications. According to a standard protocol infertility evaluation usually identifies different causes, including, male infertility (30%), female infertility (35%), the combination of both (20%), and finally unexplained or "idiopathic" infertility (15%). Female causes of infertility comprise endometriosis, tubal damage and ovulatory dysfunction (OD). Thyroid dysfunction is a condition known to reduce the likelihood of pregnancy and to adversely affect pregnancy outcome. Data on the relationship between thyroid disorders and infertility remain scarce and the association with a particular cause of infertility has not thoroughly been analyzed. In a case-control study we have shown that the relative risk of positive TPO-Abs in infertility due to a female cause and in particular related to endometriosis is significantly increased. Thyroid dysfunction itself is a condition interfering with normal ovarian function and was more frequent in women with positive anti-TPO Abs. We therefore propose that a systematic screening of TSH, free T4 and TPO-Ab could be considered in all women with a female cause of infertility. Prospective follow-up of a cohort of infertile women undergoing assisted reproduction shows a significant increased risk of miscarriage in women with positive anti-TPO Abs compared to women without thyroid auto-immunity after clinical pregnancy is established by the ART procedure. The frequent association of the presence of anti-TPO-Abs and miscarriage is hypothetical explained by the fact that organ specific autoimmune diseases may be secondary to some basic cellular abnormality that directly affects pregnancy outcome. Alternatively, women with thyroid autoimmunity, may experience greater changes in free thyroxine levels during ART and subsequent pregnancy interfering with genital tract physiology and fetal development. Determining the presence of thyroid antibodies before ART procedure is thus useful in identifying women at risk for subsequent clinical miscarriage. PMID- 12649932 TI - [Role of chemokines in the HIV infection process]. AB - In order to infect a target cell, the HIV envelope glycoprotein gp 120 has to interact with both the cellular receptor CD4 and an HIV-coreceptor, i.e. the CC or CXC chemokine receptor CCR5 or CXCR4. Both coreceptors were immediately recognized as novel targets for anti-HIV-therapy. Blocking these coreceptors would protect the cell from viral entry and would reduce the viral transmission and pathogenesis. Here we describe the purification and characterization of natural chemokine variants and compare their antiviral activity. In addition, the role of proteases for the processing of the CC chemokines RANTES, eotaxin, MDC and MIP-1 alpha and of the CXC chemokine SDF-1 are studied. The MIP-1 alpha isoform LD78 beta, that was purified form natural sources, inhibited HIV infection completely in CCR5-transfected cells, mononuclear leukocytes and purified monocytes at low (ng/ml) concentrations. This research will make it feasible to develop specific chemokine-analogs that block HIV-entry. Deciphering the processes that play a role during the complicated interactions between HIV gp120 and the cellular membrane may lead to a more efficient treatment of HIV infections. PMID- 12649933 TI - Flavobacterium columnare infections in fish: the agent and its adhesion to the gill tissue. AB - Flavobacterium columnare is the cause of columnaris disease, a serious condition affecting numerous freshwater fish species all over the world. Hitherto, only very scarce information is available on the pathogenesis of this bacterial disease, making it difficult to adopt a preventive approach to combat this pathogen. This study ambiates to shed more light on the way F. columnare interacts with its host. Since a number of difficulties occur in trying to isolate F. columnare from diseased fish, first of all, a selective medium was developed which enabled to recover various isolates from diseased live-bearing aquarium fish. In experimental infection studies in which these isolates were adopted, differences in virulence were noted. In a further stage, a clear correlation was made between virulence and the ability to adhere to the gill tissue, documenting the importance of the gills in the pathogenesis of columnaris disease. In this respect, a gill perfusion model was developed, by which means the adhesion of F. columnare to the gill tissue was partly pinpointed and various environmental factors influencing the adherence process elucidated. It may hence be stated that an important milestone has been achieved in the realisation of one of the most frequently used ways to prevent disease, that is through vaccination. Columnaris disease may also be opposed by ensuring good management practices eliciting adequate environmental circumstances. PMID- 12649934 TI - 'Nurses' dilemma--ethical issues. PMID- 12649935 TI - Role of nurses in public policy issues. PMID- 12649936 TI - Suicidal attempt: factors, seriousness & coping behaviour. PMID- 12649937 TI - Role of nursing in modern blood banking [in a tertiary level hospital]. AB - With the emergence of Transfusion Medicine Department the definition of Blood Bank has changed. Today, it is not just a bank to collect & issue blood, it prepares and supplies individual blood component, to the patients as and when the need arises during the course of treatment. The planning & management of any blood bank depends on number of functions it is required to serve and the number of units of demand of blood & blood products. This has also changed the role of nursing from skilled nursing care to managing the blood bank services. Hence, there is a need for the nursing personnel to know about the recent advances in planning and management of transfusion medicine, so that they can handle this dual responsibility as a Nurse and a manager. PMID- 12649938 TI - Mentorship in nursing. PMID- 12649939 TI - An exploratory study on gutkha and smokeless tobacco consumption. PMID- 12649940 TI - Spiritual nursing. PMID- 12649941 TI - Leeds procedure--a treatment modality for scoliosis. AB - The present study comprises of forty patients of scoliosis in growing age group (10-25 years) with Cobbs angles ranging from 52"-98". They were treated with Leeds procedure which is by anterior loosening followed by Posterior Harrington fixation + Luque derotation + Fusion and Costoplasty. Majority of the curves were thoracic (60%), and right sided (72%). Average correction of the deformity after surgery was 45%. Satisfaction level of patients and parents was good in 60% of the cases. The modality of treatment was decided on the basis of personality of each case, its demand and requirement, time of presentation and the potential for increasing severity. Anterior spinal surgery for scoliosis is an effective procedure in hands of experienced surgeons and it reduces stiffness of the curve, shorten the anterior column, decreases thoracic lordosis which leads to some improvement of pulmonary function. But there is greater risk of damage to vital structures with higher risks of cardiorespiratory failure. The potential risks have to be balanced with the expected rewards. The optimum method of correction has to be decided by careful preoperative evaluation. Combined procedure of anterior and posterior surgery causes lesser decrease in pulmonary functions than costoplasty alone, and achieving better cosmetic correction. This study reaffirms the role of anterior spinal surgery in India as the patients due to lower levels of health awareness present late and with severe deformities. PMID- 12649942 TI - Seroprevalence of Chlamydia trachomatis in women with bad obstetric history and infertility. AB - 50 sera each from women with infertility, bad obstetric history and healthy pregnant women were tested for the presence of antichlamydial IgG antibodies. Antichlamydial IgG antibodies were present in 68% of women with infertility, 50% of women with bad obstetric history and 10% in healthy pregnant women. As culture methods are cumbersome so diagnosis can be made by serological tests, which are sensitive, specific and non invasive. Chlamydia trachohmatis is one of the most common bacterial sexually transmitted disease and most infections caused by Chlamydia trachomatis being asymptomatic, result in a large reservoir of unrecognized, infected individuals capable of transmitting the infection to sexual partners. Chlamydia trachomatis if detected earlier and given proper treatment, can minimize the complications in these patients. PMID- 12649943 TI - Prevalence of anemia in Schedule Caste preschool children of Punjab. AB - A study on the prevalence of anaemia was conducted among the Scheduled Caste preschool children of Punjab. Blood sample was collected from 3,500 children ranging in age from 1+ to 5+ years and haemoglobin was estimated by cyanmethaemoglobin method. Anemia was diagnosed when haemoglobin was less than 11 g/dl. The study revealed the overall prevalence of anemia as 81.66%. The frequency of anemia was maximum in age group 2+ and minimum in age group 5+. PMID- 12649944 TI - Burden of chronic severe anemia in obstetric patients in rural north India. AB - There were 5124 deliveries during January to December 2000 and of these 386 patients (7.53%) were severely anemic (Hb < 6.0 g/dl). 1083 units of blood were transfused to these patients, a mean of 2.80 units to each patient. There were 2 maternal deaths due to severe anemia. Of the 386 patients, 246 could be interviewed in detail and their demographic details were compared with 100 normal pregnant women with haemoglobin levels of 10 g/dl or more. In the anemic group 81.7% were illiterate, 62.1% had no antenatal check, 78.8% had no access to safe drinking water 90.65% had no toilet facilities and used open fields for defaecation, 63.1% reported having diarrhoea and pain in abdomen off and on. 20.73% reported history of passage of worms in the stools. History of previous menorrhagia was present only in 1.2%. The comparable figures in the non-anaemic group were 34% illiterate, 24% no antenatal checkup, 55% no toilet facilities, 38% unsafe drinking water, 6% diarrhoea, 2% (worms in stools) and 5% menorrhagia. Hence it is concluded that along with prophylaxis with iron and folic acid sufficient consideration should be given to sanitation facilities, safe drinking water, recurrent gastrointestinal infection etc. PMID- 12649945 TI - Translocation Down syndrome. AB - Cytogenetic investigations carried out on 1021 cases of Down syndrome revealed translocation in 46 cases. The most frequent was of t(14;21) and t(21;21) types. Most of the translocation DS cases (n = 31) were born to younger mothers (< 25 years), when compared to pure trisomy 21 DS cases. Parental karyotypes, family history and parental ages has helped us greatly in offering genetic counseling, prenatal diagnosis and estimating the risk for the next conception. PMID- 12649946 TI - Effect of vitamin A supplementation on childhood morbidity and mortality. AB - In a double blind design, 1520 children aged < 10 years were individually randomised in vitamin A and placebo group in slums of Chandigarh. Children > 12, 6-12 and < 6 months of age received 200,000, 100,000, 500,000 I.U. of vitamin A respectively every 4 to 6 months during 15 months trial period. The prevalence of vitamin A deficiency was significantly reduced in vitamin A compared to placebo group during the follow-up period. In vitamin A group, incidence of diarrhoea and measles was significantly reduced but incidence of acute respiratory infections was not significantly different compared to control group. Risk of death was also significantly less in vitamin A group. Therefore, promotion of vitamin A rich diet or supplementation with synthetic vitamin A at 4-6 month interval should be a priority in populations where risk of vitamin A deficiency is high. PMID- 12649947 TI - Electrophoretic analysis of Salmonella typhi and other bacteria. AB - A comparative study involving SDS-PAGE of Salmonella typhi and other Bacteria was conducted. Protoplasmic antigens of Salmonella typhi. Salmonella paratyphi A, Salmonella typhimurium, Proteus sp, Klebsiellas sp. Pseudomonas aeruginosa, Shigella flexneri, Staphylococcus aureus were separated and compared on SDS-PAGE followed by checking of their cross reactivity by gel diffusion using antisera raised against whole cell and lysates of Salmonella typhi. Lines of identity between Salmonella typhi, Salmonella paratyphi A and Salmonella typhimurium were observed. No lines of identity were seen among Salmonella typhi, Pseudomonas aeruginosa and Sfaphylococcus aureus. PMID- 12649948 TI - Scleredema diabeticorum--a case report. AB - A 41 year male, diabetic on treatment presented with persistent erythema and. "taut" skin over neck and back since 1 year. The lesions showed symmetric and gradual progression. There was no history of prior sore throat. On examination effected skin was erythematous, woody hard and unpinchable. Scleredema diabeticorum (diutinum), although sharing clinical and historical features with Scleredema adultorum has no prodromal infection, is more extensive and affected individuals are characteristically obese, middle aged diabetics who often have accompanying microangiopathies and macroangiopathies. Recognition of scleredema by the physician has prognostic and therapeutic implications in the management of the coexisting diabetes. PMID- 12649949 TI - Enteric fever due to Salmonella Weltevreden in a four-year-old child. AB - A four-year old child was admitted with signs and symptoms suggestive of enteric fever. Blood culture and serial stool cultures were undertaken. Weltevreden, a rare Salmonella serotype was isolated from the stool samples. The isolate was sensitive to ampicillin, cefotaxime, gentamicin, chloramphenicol and ciprofloxacin. PMID- 12649950 TI - Antimicrobial activity of extracts of Terminalia catappa root. AB - The effect against bacteria of petroleum ether (60-80 degrees C), chloroform and methanolic extract of dried root of Terminalia catappa Linn. (combrataceae) was employed by cup plate agar diffusion method. The chloroform extract showed prominent antimicrobial activity against S. aureus and E. coli as compared to other tested microorganisms, while petroleum ether extract was devoid of antimicrobial activity. The methanolic: extract exhibited MIC of 0.065 mg/ml against E. coli. and chloroform extract exhibited MIC of 0.4 mg/ml against S. aureus The chloroform has well as methanolic extracts showed good antimicrobial activity against Gram positive and Gram negative microorganisms. PMID- 12649952 TI - [Use and reliability of the Delvo-P-test]. AB - The Delvo-P-test (Gist-brocades nr, Delft, The Netherlands) is based on biological detection of antibiotics in the milk. According to the procedure of the manufacture the test is suited for bulk milk processing factories. Problems arise when the test is used on milk samples of a single cow. Buffers, cells, bacteria, cell contents, and desinfection without using antibiotics may give false positive and false negative results. An error up to 44.8% was detected on single cow milk samples. The test is well suited for bulk milk, where the processing of milk follows a biological procedure as for cheese. The use on single cow milk samples to prove absence of antibiotics is not recommended. PMID- 12649951 TI - [Porcine malignant catarrhal fever: diagnostic findings and first detection of the pathogenic agent in diseased swine in Switzerland]. AB - For the first time Ovine Herpesvirus 2 (OvHV-2) was identified in Swiss pigs as the causative agent of Porcine Malignant Catarrhal Fever (MCF). Diseased animals from two farms were observed to show weakness, anorexia, fever up to 41 degrees C, and neurological symptoms, i.e. ataxia, convulsions and hyperesthesia, erosion on the snout and in the oral and nasal mucosa, as well as multiple skin lesions. Histopathological findings included severe non-purulent inflammation with mononuclear cell infiltration in several organs. Most dominant were meningo encephalitis, disseminated nephritis as well as purulent catarrhalic bronchopneumonia. The findings were quite reminiscent of the lesions due to MCF in cattle and give therefore substantial proof to use Porcine Malignant Catarrhal Fever as the term for the disease. Identification of the causative agent was done with a quantitative PCR specific for OvHV-2. Different tissues from diseased animals were positive. Furthermore, one animal which had been ill for more than five days tested positive for antibodies against an epitope conserved among MCF viruses. Serum samples from diseased animals reacted negative towards Classical Swine Fever- and Pseudorabies virus antigen. A weakly positive reaction against porcine enterovirus type I argued against the involvement of enteroviruses in the observed disease. Moreover, by means of different conventional PCRs, we detected the newly discovered porcine lymphotropic herpesviruses for the first time in Switzerland and could at the same time exclude their involvement in Porcine Malignant Catarrhal Fever. PMID- 12649953 TI - [Validation of DNAse reaction for identification of Staphylococcus aureus strains in routine mastitis diagnosis]. AB - In routine mastitis diagnostic S. aureus strains are often identified using DNAse reaction. However, this enzyme activity is not limited only to S. aureus strains. Furthermore, the strength of the DNAse reaction between different strains varies strongly. These factors lead to the fact that the results are often not comparable between laboratories. The aim of these investigations was to validate the DNAse reaction for routine identification of S. aureus in mastitis diagnostic and to set a critical limit for interpretation of DNAse reaction. The results of 189 strains isolated from bovine mastitis milk samples show that colonies with beta delta or delta-haemolysis and DNAse reactions of > or = 2 mm can reliably be identified (sensitivity 91.2%, specificity 96.9%) as S. aureus. PMID- 12649954 TI - [Cardiac tamponade due to pericardial mesothelioma in an 11-year-old dog: diagnosis, medical and interventional treatments]. AB - In a dog presenting with the clinical signs of exercise intolerance and ascites, cardiac tamponade due to suspected idiopathic pericarditis was diagnosed based on thoracic radiographs, electrocardiogram (EKG) and cardiac ultrasound. Pericardial effusion recurred soon after pericardiocenteses, prescription of colchizine and again after balloon pericardiotomy. After partial pericardectomy by thoracoscopy and after obtaining a histological diagnosis of mesothelioma adjuvant intracavitary chemotherapy using cisplatin was performed. Already one week later the dog developed marked dyspnea due to severe pleural effusion. The dog was maintained at acceptable life quality judged based on playfulness and appetite using repeated pleuro-centeses for an additional two months, when the dog was euthanized due to uncontrollable pleural effusion. Despite extensive treatments life span from initial presentation to euthanasia was only 5 months. Necropsy revealed extensive mesothelioma metastases covering the whole pleura, epicardium and remaining pericardium. Diagnostic and therapeutic aspects of (recurrent) pericardial effusion are discussed based on this case. PMID- 12649955 TI - [Metastases of perianal gland carcinoma in a dog: palliative tumor therapy]. AB - Palliative radiation therapy may provide significant relief in symptoms in pets with incurable cancer. Therapy is applied over a short period of time, using larger than normal fractional doses. Palliative radiation leads to minimal or no side effects. This case report describes the situation of a miniature poodle that was presented with severe dyschezia as well as fresh blood in the feces. The dog had a large abdominal mass which was diagnosed as lymph-node metastasis of a perianal gland carcinoma. Therapy included palliative radiation as well as chemotherapy. Six months after initial presentation the dog is free of clinical symptoms. PMID- 12649956 TI - [Interactive workshops as a dissemination strategy in psychology]. AB - OBJECTIVE: To assess whether interactive workshops are an effective strategy for promoting a psychological intervention model among healthcare providers, to treat problem drinkers. MATERIAL AND METHODS: The study was conducted between the years 1999 and 2000, among 206 healthcare providers at seven Instituto Mexicano del Seguro Social (Mexican Institute of Social Security, IMSS) clinics. Study subjects were selected by hospital executive officers. The study design is a quasi-experimental pre-test/post-test study. Data on providers' attitudes, interests, and knowledge were collected using a questionnaire. After that, interactive workshops were conducted, and the same questionnaire was applied again at the end of the workshops. Statistical analysis was carried out using Student's t test for matched samples. RESULTS: Statistically significant differences were found in participants' knowledge on alcoholism t (206, 205) = 9.234, p = 0.001, as well as in their interest t (206, 205) = -2.318, p = 0.021. CONCLUSIONS: Interactive workshops are an effective tool to disseminate the Guided Self-Help Program conducted in IMSS clinics. Healthcare providers can become change-inducing/promoting agents of psychological innovations. PMID- 12649957 TI - Emotional dysfunction associated with diabetes in Mexican adolescents and young adults with type-1 diabetes. AB - OBJECTIVE: To assess the emotional dysfunction associated with diabetes in Mexican young individuals with type-1 diabetes. MATERIAL AND METHODS: A cross sectional survey was conducted to perform a complete clinical and psychosocial evaluation of 93 consecutive type-1 diabetes patients, and 14 to 30 years. RESULTS: Adolescents had higher scores of emotional dysfunction related to diabetes and a diminished knowledge in diabetes-related areas. A multivariate logistic regression model showed that an inadequate emotional response to diabetes (high problem areas in diabetes or PAID scores) was mainly associated with a poor glycemic control (OR = 2.9, 95% CI 0.9-9.7, p = 0.09). Apprehension about the future and the possibility of serious complications had the highest mean PAID score in all age groups. CONCLUSIONS: New strategies should be developed to improve the routine care and support of young individuals with type 1 diabetes. PMID- 12649958 TI - [A model for integrating mental health care resources in the rural population of Mexico]. AB - OBJECTIVE: The purpose of this project was to identify behaviors of health service utilization to solve mental health-related problems among rural inhabitants of Mexico. A model of pathways to mental health services was built. Based on this model, an integration-intervention model is proposed to help improve access to mental health services and their quality. MATERIAL AND METHODS: In 1997, in-depth interviews were conducted among 21 adults (9 men and 12 women) and seven healthcare providers, in a rural community in Jalisco, Mexico. Women's age ranged from 23 to 44 years; ten were married and two single. Men were between 30 and 74 years old; eight were married and one was a widower. Healthcare providers in the same community were: a priest, a general physician, a pharmacy clerk, two nurses, and two traditional healers. RESULTS: The pathways models suggest that the first attempt a person does to solve a symptom is self-care. When such strategies are not sufficient to relief the symptom, the person turns to the members of his/her social network for help, who in addition to providing information about remedies, offer their emotional and instrumental support. If after consulting the social network, the symptom is not relieved, the individual seeks help from other external resources, such as the members of the ethnomedical local system. Inhabitants of rural communities tend to seek help from physicians, only when the symptom persists and the suffering associated with it seems to be out of the individual's control, or if members of the social network or the ethno medical local system refer the patient to the physician. Seeking help from a specialist in mental health (psychiatrist or psychologist) is quite improbable among the rural inhabitants of Mexico, mostly due to difficulties in accessing these services, such as geographical location, distance, transportation, cost, and cultural distance between the health providers and the patient. CONCLUSIONS: In order to conduct successful intervention programs that are culturally sensitive for rural inhabitants, it is necessary first, to have detailed information derived from research and second, the active participation of all human and institutional resources, from professionals and truly multi disciplinary researchers, to educators, social, religious, political leaders, and members of the communities and local institutions. The proposed model of integration of personal and community resources for healthcare requires establishing programs that sensitize, inform, and train community members, as well as members the ethno-medical system and the formal heath system. PMID- 12649959 TI - Costs of publicly provided maternity services in Rosario, Argentina. AB - OBJECTIVE: This study estimates the costs of maternal health services in Rosario, Argentina. MATERIAL AND METHODS: The provider costs (US$ 1999) of antenatal care, a normal vaginal delivery and a caesarean section, were evaluated retrospectively in two municipal hospitals. The cost of an antenatal visit was evaluated in two health centres and the patient costs associated with the visit were evaluated in a hospital and a health centre. RESULTS: The average cost per hospital day is $114.62. The average cost of a caesarean section ($525.57) is five times greater than that of a normal vaginal delivery ($105.61). A normal delivery costs less at the general hospital and a c-section less at the maternity hospital. The average cost of an antenatal visit is $31.10. The provider cost is lower at the health centre than at the hospital. Personnel accounted for 72-94% of the total cost and drugs and medical supplies between 4-26%. On average, an antenatal visit costs women $4.70. Direct costs are minimal compared to indirect costs of travel and waiting time. CONCLUSIONS: These results suggest the potential for increasing the efficiency of resource use by promoting antenatal care visits at the primary level. Women could also benefit from reduced travel and waiting time. Similar benefits could accrue to the provider by encouraging normal delivery at general hospitals, and complicated deliveries at specialised maternity hospitals. PMID- 12649960 TI - [Alimentary basic basket and index of prices in Santander, Colombia, 1999-2000]. AB - OBJECTIVE: To establish indicators for food security (FS) in two Colombian municipalities. MATERIAL AND METHODS: In 1999-2000, a descriptive study was carried out in two municipalities of the department of Santander, Colombia, that determined the cost of basic food baskets (BFB), to estimate price indices (PI), their variation, and indicators of FS related to the legally set minimum wage (MW). RESULTS: No differences were found in the PI by municipality (p > 0.05). The annual cumulative price indices were a single digit. The percentages of food insecurity (FI) were upwards of 50%, differing by municipality, in 1999 (p = 0.04), and 2000 (p = 0.88). The FI increased five points on average for the period 1999-2000. An average minimum wage of 1.24 per month is needed for a family to have access to a BFB. CONCLUSIONS: The purchasing power of the current MW doesn't satisfy the caloric nor nutritional requirements of a family. The local FS will continue deteriorating, given the behavior of its determinants. PMID- 12649961 TI - [Determinants of dengue transmission in Veracruz: an ecological approach to its control]. AB - OBJECTIVE: To assess the ecological, social, and demographic factors associated with the transmission of dengue virus infection in Veracruz, Mexico, and to identify risk areas to target control measures. MATERIAL AND METHODS: This ecological study included data for 1,249 localities within the 11 Health Jurisdictions of the State of Veracruz, Mexico, for the 1995-1998 period. The following data were collected for each locality: total number of cases per year, population by sex, number of households, provision of public services, altitude, latitude, longitude, and deprivation index. RESULTS: Dengue transmission was registered in 17% of the localities in the State; 70% of the cases were concentrated in only 6% of the localities. Recurrent localities were urban centers with adequate availability of public services and low deprivation indices. Dengue transmission was detected in rural areas, but it was not common. The average number of cases differed according to the size of the locality and the number of years dengue was reported. A population threshold to maintain transmission was found. CONCLUSION: Recurrent localities concentrating 70% of dengue fever cases were identified, as well as the ecological and demographic factors associated with dengue transmission. The risk stratification approach to dengue transmission may improve control and prevention of this disease in high risk areas. PMID- 12649962 TI - [Results of a survey on organ donation and transplantation in the metropolitan area of Guadalajara, Mexico]. AB - OBJECTIVE: To evaluate knowledge of and disposition towards organ and tissue donation for transplantation among the adult population of metropolitan Guadalajara, Jalisco, Mexico. MATERIAL AND METHODS: A survey was conducted by personal interview with street-recruited persons over 18 years of age using a 29 item questionnaire. Metropolitan Guadalajara was divided into four municipalities and a proportional number of interviews was assigned to each one based on number of inhabitants, socio-economic stratum, age and gender. RESULTS: Four hundred individuals were interviewed. Approximately 80% knew about organ donation and 65% knew that it was legal; however, only 24% had discussed the topic with their relatives. Seventy-one percent did not know how to donate organs and only 25% knew about organ donor cards. On the other hand, 66% were willing to donate, 16.5% would not donate and 17% were undecided. Bodily mutilation was the main reason (40%) given to not donate. 89% would authorize organ donation from a deceased relative, assuming the relative had previously expressed the desire to donate; on the contrary, only 29% would give such an authorization without their relative's prior consent. Finally, 65% considered the driver's license an adequate mean to express their desire to donate and 50% considered the information the mass media disseminated on the subject to be insufficient. CONCLUSIONS: The majority of the metropolitan Guadalajara population knew about organ donation and would be willing to donate their organs. However, a high percentage did not know how to donate and considered the information disseminated on the subject insufficient. An on-going mass media campaign could resolve this matter and hopefully increase organ donation among this population. PMID- 12649963 TI - Coordination of international multicenter studies: governance and administrative structure. AB - A well-conducted multicenter study needs to assure standardization, uniformity of procedures, high data quality, and collaboration across sites. This manuscript describes the organization and dynamics of multicenter studies, focusing on governance and administrative structures among countries of diverse cultures. The organizational structure of a multicenter study is described, and a system for oversight and coordination, along with roles and responsibilities of participants in the multicenter study, are presented. The elements of a governance document are also reviewed, along with guidelines and policies for effective collaboration. The experience of an ongoing multi-country collaboration, the World Studies of Abuse in the Family Environment (WorldSAFE), illustrates the implementation of these guidelines. It is essential that multicenter studies have an objective coordinating center and that the investigators jointly develop a written governance document to enable collaboration and preserve collegiality among participating investigators. PMID- 12649964 TI - [What is your ultrasonic diagnosis? Infected urachus apertus]. PMID- 12649965 TI - [Reference values in pregnancy]. AB - The body of pregnant women demonstrates in many ways different functions compared to non pregnant women. In order to assess lab results of antenatal tests the knowledge of physiologic and pathophysiologic changes are essential. This article reviews the state of the art in terms of reference values during pregnancy enriched with own results. Of special value for the practitioner are such reference values and curves for the circulatory system, kidney function, hematologic changes, plasma components and weight gain. PMID- 12649966 TI - [Post-traumatic stress disorders: diagnostic and therapeutic principles ]. AB - Over 50 percent of the general population are exposed to a traumatic event at least once in their lives. Some but not all trauma survivors go on to develop posttraumatic stress disorder (PTSD). The diagnostic criteria of acute stress reaction, posttraumatic stress disorder, and enduring personality change after catastrophic experience are presented. PTSD should primarily be treated by means of psychotherapy. Cognitive-behavioral approaches, exposure therapy in particular, currently show the best evidence as effective treatments for PTSD. In addition to psychotherapy, psychopharmacological treatment may be indicated. Selective serotonin-reuptake inhibitors are the first-line psychoactive drugs for PTSD. Due to an increased risk of substance use disorders, benzodiazepines should only be used with caution. Finally, a number of predictors are reported that may help the clinician to identify patients at risk of developing PTSD. PMID- 12649967 TI - [Left-sided chest pain]. PMID- 12649968 TI - [Persistent fatigue despite sufficient sleep]. PMID- 12649969 TI - [The extraction of truth: apropos of the Socratic dialogue]. AB - The socratic dialogue is a philosophical method that enables colleagues to investigate which judgements people have about their experiences and how these judgements can be based. In this article, the reader will learn more about the historical background, the organisation, the levels of dialogue, the role of the facilitator. We also pay attention to the results that a regular practise of socratic dialogue can have for professional dentists. The most important one is a growing sensitivity and lucidity in the daily social life with patients and colleagues. In the dialogue, this can be practiced by sharpening the moral perception of concrete details in the lived experience. PMID- 12649970 TI - [Who would suffer this child in the cold? History and diagnosis during a Socratic dialogue]. AB - This article gives an account of the experiences of 12 dentists (11 general practitioners and one periodontist) during a one-day Socratic dialogue, which took place at the International Conference for Dentists in Bruges 2001. The key question was: "When must I (or mustn't I) comply with the patient's wish?" The participants tackled an example from Edgar, general practitioner. His story went like this: on Christmas Eve, dinnertime, one of his patients--duly instructed beforehand--arrived just like that, and said: "I'm in pain". When the central question was applied to this example, uncertainty arose about the exact nature of "the patient's wish". In this article you read how the participants dealt with the "pain", with the uncertainty. PMID- 12649971 TI - [Woman and career: an unhappy marriage? A study model: dentistry]. AB - Women's participation in the professions is steadily increasing. This study is a quantitative assessment of male and female career patterns in dentistry and of the role of marriage in any gender gap. A 1997 survey explored the professional activity of Flemish dentists. There is no educational disparity between dentists, female dentists enter the profession on an equal footing with their male colleagues. Therefore any other career influencing factors reveal themselves relatively clearly. Dependent variable was income; independent variables were gender and marriage. Female dentists scored lower than male dentists in each career phase; marriage made no significant difference. However, marriage turned out to be an important positive factor in the career of male dentists. Women dentists marry significantly later and less than their male colleagues. Further research, also of a qualitative nature, is necessary to evaluate the professional and social impact of this substantial gender difference. PMID- 12649972 TI - [25 years of INAMI data in dentistry]. AB - The author presents 25 years of social security data about expenses and the number of medical acts in dentistry using lists and diagrams. By relating these expenses per year to the number of dentists per year, the average expenses per dentist are compared to the evolution of the index of consumption. In the same manner the average number of medical acts per dentist are calculated. By this method the author emphasizes the changes in average dental practice profile over 25 years. PMID- 12649973 TI - [Criminal enquiry and determination of dental damage. Analysis of a prosthetic fracture following an accident]. AB - When signing a legal insurance policy, one is not always aware of the seriousness of the reciprocal legal consequences. Physical injuries mainly involve civil procedures in spite of the fact that criminal proceedings are instituted in some cases. This article deals with the judicial inquiry concerning the notification of an accident with insurance claim. The object of this inquiry is a porcelain fracture caused by a so-called accident where verification becomes very complicated for the appointed experts. If diagnosis, anamnesis and the initial examination indicate possible fraud in the case, the expert's assessment for the insurer will rightly include the criminal section. What does this involve for the expert and his team? The violation of the public interest by making a false statement will result in the expert trying to reveal the truth instead of estimating the damage. PMID- 12649974 TI - [Indications for orthognathic surgery, the limitations of orthodontics and of surgery]. AB - The different sagittal, vertical or transversal skeletal dysmorphosis can be treated with orthodontic or orthopaedic appliances. Certain limits of treatment will be due to the age of the patient, the importance of the skeletal dysmorphosis, technical limits of the orthodontic appliances or the motivation and compliance needed from the patient. For this reason an orthodontic treatment combined with orthognathic surgery may be indicated. Nevertheless the combined orthodontic-surgical treatment has also its limitations and specific treatment needs from orthodontist and maxillo-facial surgeon. PMID- 12649975 TI - [A multidisciplinary approach to orthognathic surgery]. AB - Orthodontists are precisely aware of their therapeutic limits in dealing with maxillofacial dysmorphia. Unstable results and iatrogenic lesions caused by alveolo-dental compensations for displacements of the skeletal base have convinced them of the importance of orthognathic surgery as a complement to their orthodontic treatments. Collaboration between the orthodontist and the maxillofacial surgeon begins as of the moment a displacement of the skeletal bases is established through clinical and cephalometric diagnosis. Due to factors related to their age and dental history, it is necessary to complement the therapeutic provision for adult patients with the provision of a multidisciplinary team. The dentist and the family doctor take part in the decision regarding the treatment to be administered, they provide advice to their patients and treat them according to the chosen therapeutic sequence. Other practitioners are involved, including the physiotherapist or speech therapist, depending on the clinical requirements. This multidisciplinary approach and collaboration are conducive to the quality of the results and patient satisfaction. PMID- 12649976 TI - [The psychological implications of orthognathic surgery]. AB - The orofacial area is strongly connected with one's self image and therefore, plays a role in social interaction. Most orthognathic surgery gives rise to functional and esthetic changes that are assumed to render physical and psychological benefit. Despite the objective physical benefits, unexpected psychologic or emotional effects may occur. This short article highlights some of those aspects. PMID- 12649977 TI - [Pre- and post-surgical orthodontic objectives and orthodontic preparation]. AB - Orthodontics in combination with orthognathic surgery is a possibility today for correcting sagittal, vertical and transversal dysmorphosis. To do this, orthodontic preparation before surgery is necessary. In the sagittal plane, dental compensations should be removed. In the vertical plane, levelling of a severe mandibular SPEE curve should be accomplished postsurgically, and transverse coordination obtained. Orthodontic preparation is very specific to the type of dysmorphosis: class II malocclusion, class III malocclusion or in open bite. Postsurgical orthodontic treatment is used to finalize tooth alignment as in conventional orthodontics. PMID- 12649978 TI - [Surgical technics in orthognathic surgery]. AB - The goal of this paper is to explain simply the most used surgical technics in orthognathic surgery. The author insists upon the noble structures met during this surgery. He also insists upon the modifications of the facial mask by the mobilisation of the different parts of the facial skeleton. Numerous schemas illustrate and make easy the comprehension of this paper. PMID- 12649979 TI - [Complications and failures in orthognathic surgery]. AB - This paper reviews the literature about the most relevant complications in daily orthognathic surgery. Although orthognathic surgery has proven to be relatively safe, patients who undergo such maxillofacial orthopaedic surgery must be aware of the side effects of this combined treatment. Immediate life-threatening complications are very rare. They can in most cases be avoided by good anaesthetic and surgical techniques and adequate postoperative care. The most frequently encountered perioperative problem in maxillary surgery is excessive blood loss, whilst subjective neurosensory disturbance is the most frequent complication in mandibular surgery. Good co-operation between orthodontist and surgeon is essential to prevent most immediate and late postoperative problems and nearly all unsatisfactory results. PMID- 12649980 TI - [The role of maxillofacial surgery in obstructive sleep hypopnea and apnea syndrome]. AB - One of the most common symptoms of obstructive sleep apnea syndrome (OSAS) is daytime drowsiness. It is associated with a high cardiovascular morbidity and mortality, and an elevated incidence of car crashes. In general, young patients don't want conservative treatment because symptomatic and to prevent secondary effects. In this article, we briefly define sleep disorders and the interest of cephalometric examination. We describe the different treatment possibilities and stress the important role of orthognathic surgery in this syndrome. PMID- 12649981 TI - [The connection between orthognathic surgery and the temporomandibular joint]. AB - The effects of orthognathic surgery on the temporo-mandibular joint constitute a major problem with orthognathic surgery largely due to their unpredictability. A review of the literature of these consequences shows little consensus among the authors and this is probably because of a lack of precise details for the criteria used. After having analysed the susceptible mechanisms to explain the pathologies of the temporo-mandibular joint during orthognathic surgery, this article proposes a protocol for the management of these difficulties. PMID- 12649982 TI - [The Association for Parents of Children born with Cleft Lip and/or Palate]. PMID- 12649983 TI - [The Association for Congenital Facial Dysmorphoses. " The face in perspective"]. AB - The aims and work of the volunteer organisation "Association of congenital facial dysmorphose" is described. Follows a summary of a book which includes a very large approach to the problem. Medical-technical contributions are completed with testimonies and practical (pedagogical) tips. PMID- 12649984 TI - [Bone distraction osteogenesis]. AB - Bone osteogenesis has been used by many surgeons to compensate dissymmetry due to congenital malformations, diseases or traumas. Bone distraction is the process of generating new bone in a gap between two bone segments in response to the application of graduated tensile stress across the bone gap. Different types of distraction are actually proposed to be an alternative to facial osteotomies or bone grafting techniques. Distraction can be unilateral, bilateral and correct the deficiencies in one, two or three dimensions. The advantages of the distraction osteogenesis are the possibilities to obtain bone lengthening together with simultaneous expansion of the functional soft tissue matrix including skin, muscles, nerves, cartilage, blood vessels and periosteum. Distractors are internal or external devices made of pins or miniplates fixed on the bone and connected by lengthening--screw--systems allowing elongation. PMID- 12649985 TI - [Diagnostic imaging of calculi in the upper urinary tract--sonography vs. computerized tomography]. AB - The choice of the most suitable technique for radiologic evaluation of upper urinary tract stones depends on the precise clinical questions that have to be answered. Many of these questions can still be answered by plain films of the abdomen and excretory urography. This review addresses the value of ultrasonography and computed tomography (CT) with respect to the two most important clinical questions, i.e. 1) presence/extent of obstruction/hydronephrosis and perinephric abscess in patients with acute flank pain; and 2) precise location, number and size of calculi. Because its sensitivity is comparable with CT and it is widely available, ultrasonography in the hands of experienced clinicians/radiologists may be preferred for evaluation of patients with acute flank pain. However, it must be emphasized that ultrasonography may totally miss acute ureteral obstruction/hydronephrosis within the first 12-24 hours. In children as well as in pregnant women, ultrasonography is still the technique of choice, but it may be replaced by magnetic resonance urography in the future. For precise stone location or detection of calcifications, however, the speed, safety and accuracy of unenhanced helical CT make this the most sensitive method and therefore the technique of choice. It also detects urinary calculi more accurately and exposes patients to less radiation than the traditional combined plain abdominal film/intravenous urography. Furthermore, CT can most readily reveal alternative diagnoses in patients with acute flank pain and other intraabdominal pathologies than stones. PMID- 12649986 TI - [Pathophysiology, diagnosis and conservative therapy in calcium kidney calculi]. AB - Annual incidences of kidney stones are about 0.1-0.4% of the population, and lifetime prevalences in the USA and Europe range between 8 and 15%. Kidney stones occur more frequently with increasing age and among men. Within ten years, the disease usually recurs in more than 50% of patients. Nowadays, about 85% of all kidney stones contain calcium salts (calcium oxalate and/or calcium phosphate) as their main crystalline components. Because human urine is commonly supersaturated with respect to calcium salts as well as to uric acid, crystalluria is very common, i.e. healthy people excrete up to ten millions of microcrystals every day. Recurrent stone formers appear to excrete lower amounts or structurally defective forms of crystallization inhibitors which allows for the formation of large crystal aggregates as precursors of stones. Alternatively, crystal adhesion to urothelial surfaces may be enhanced in stone formers. Medical treatment of renal colic is based on nonsteroidal antiinflammatory drugs, because prostaglandins appear to play a crucial role in the pathophysiology of pain during ureteral obstruction. In addition, centrally acting analgesics such as pethidine-HCl may be required in many cases. The administration of high amounts (3-4 liters/day) of intravenous fluids should be abandoned, since it may raise intraureteral pressure whereby pain increases and kidney pelvis or fornices may rupture. All first-stone formers should undergo a simple basic evaluation, including stone analysis (x-ray diffraction or infrared spectrometry), serum values of ionized calcium (alternatively: total calcium and albumin) and creatinine, urinalysis and repeated measurements of fasting urine pH in order to detect urinary acidification disorders or low urine pH. In high-risk patients with as first stone episode (i.e. strongly positive family history, inflammatory bowel disease, short-bowel syndrome, nephrocalcinosis, bilateral stones, hypercalcemia, renal tubular acidosis, airline pilots) as well as in all recurrent stone formers, an extended metabolic evaluation should be performed. Two 24-hurines should be collected on free-choice diet not prior to three months after stone passage or urological intervention. Analysis includes measurements of volume, creatinine, calcium, oxalate, uric acid and citrate; sodium and urea as markers of salt and protein consumption are optional but clinically very helpful. Since hypercalciuria is of much less importance than increases in urinary oxalate, therapeutic efforts should primarily focus on lowering urinary oxalate excretion. Sufficient calcium intake, i.e. 1200 mg per day, is crucial, because it allows for binding of oxalate at the intestinal level whereby increases of urinary oxalate (reciprocal hyperoxaluria) can be avoided. Excess intake of flesh protein (meat, fish, poultry) is lithogenic since it increases urinary calcium, oxalate and uric acid, and lower citrate. On the other hand, a diet rich in alkali (vegetables, fruit) is associated with a lower risk of stone formation. A "common sense diet" containing sufficient amounts of fluids, 1200 mg of calcium per day and reduced amounts of flesh protein as well as salt is able to reduce the 5-year stone recurrence rate in calcium stone formers by 50%. The scientific evidence for drug treatment (thiazides, alkali citrate) is rather poor: the most widely quoted randomized thiazide trial included only 42 patients of whom 36% left the protocol prematurely, whereas 36-48% of patients included in three randomized studies with alkali citrate suffered from undesirable side-effects; nevertheless, citrate therapy reduced the stone recurrence rate by 38%, compared with 22% in patients on placebo treatment (p < 0.0005). PMID- 12649987 TI - [Pathophysiology, diagnosis and conservative therapy of non-calcium kidney calculi]. AB - While calcium oxalate and calcium phosphate make up at least 80% of all kidney stones, infection-induced and uric acid stones occur in 10% and 8%, respectively. Although any type of stone may become infected, the term "infection stones" means that stone formation exclusively depends on urease-producing bacteria. The splitting of urea leads to a rise in urinary pH which may induce crystallization of struvite (magnesium-ammonium-phosphate), the major constituent of infection stones, or carbonate apatite. Struvite stones account for the majority of staghorn calculi. They can grow quite large and may fill the entire collecting system. Patients with struvite stones may present with acute flank pain or remain completely asymptomatic. The cure of infection stones requires complete removal of the stone material. For uric acid crystallization and stone formation, low urine pH (below 5.5) is a more important risk factor than increased urinary uric acid excretion. Main causes of low urine pH are tubular disorders (including gout), chronic diarrheal states or severe dehydration. Accordingly, the treatment of uric acid stones consists not only of hydration (urine volume above 2000 ml per day), but mainly of urine alkalinization to pH values between 6.2 and 6.8. Urinary uric acid excretion can be reduced by a low-purine diet as well as--in case of recurrent uric acid stones and/or gout--by allopurinol. Cystinuria is a rare hereditary gene disorders with impaired tubular reabsorption of cystine. Stone formation occurs as a consequence of cystine's relatively low solubility at urine pH levels below 8. Only symptomatic diet and drug treatments are currently available, with urine dilution and urine alkalinization being the most efficient ones. Cystine stones respond poorly to shockwave lithotripsy, so that invasive procedures may regularly be necessary. 2,8-dihydroxy-adenine stones occur as a consequence of an enzyme deficiency that involves purine metabolism. These resulting stones are not visible by fluoroscopy and are therefore often misinterpreted as uric acid stones. Low-purine diet and allopurinol reduce the frequency of stone formation. PMID- 12649988 TI - [Urologic interventional therapy of kidney calculi (I)--extracorporeal shockwave lithotripsy]. AB - With a prevalence of around 5% in western countries, urolithiasis is a frequently occurring disease but with a poorly understood pathogenesis. Effective prevention is not possible for most stone types, in particular for calcium-containing stones, which occur most frequently. Additionally, after occurrence, patient compliance is often inadequate. Interventional therapy becomes necessary after stone manifestation within the urinary tract. Not all stone compositions respond to a drug treatment. Whereas in the seventies, stone treatment meant open surgery newly developed minimally-invasive procedures have displaced this treatment. The clinical introduction of extracorporal shock wave lithotripsy (ESWL) has played a crucial role in this process. Today, more than 80% of all urinary stones can be treated by modern lithotripters. Combination with other minimally-invasive procedures further improve stone free rate. This article provides an overview of technique, application and results of ESWL treatment. PMID- 12649989 TI - [Urologic interventional therapy of kidney calculi (II)--Endoscopic methods: ureterorenoscopy and percutaneous nephrolitholapaxy]. AB - Although ESWL allows for the treatment of most urinary stones, there are remaining situations, in which satisfying results cannot be achieved, e.g. large kidney stones or specific anatomy. In such cases endoscopic procedure like ureterorenoscopy (URS) or percutaneous nephrolithotomy (PCNL), alone or in combination with ESWL, allow minimally-invasive removal of these stones. Primarily, the development of modern intracorporeal lithotripter probes and extraction instruments along with a reduction of instruments' diameter could improve efficacy and reduce mortality. This article will provide an overview of techniques, application and results of endoscopic urinary stone treatment. PMID- 12649990 TI - [Gallstones--natural history and conservative management]. AB - Gallstones have a very high prevalence affecting 9.5 and 19.5% of men and women, respectively. The pathophysiology and risk factors for cholesterol gallstones are considered. Modern imaging techniques, in particular ultrasound, are very sensitive to detect cholecystolithiasis, while ERCP remains the gold standard to detect choledocholithiasis. ERCP could be supplanted soon by endosonography and MRCP. Biliary scintigraphy is useful to detect a 'vesicule exclue' and to document gallbladder/sphincter Oddi dysfunction. The different conservative management strategies including litholytic treatment with ursodeoxycholate and lithotripsy have been largely abandoned in favor of laparoscopic cholecystectomy. Litholytic treatment has still a role to play in the prevention of gallstone formation in patients with rapid weight loss and in the newly detected MDR3 defect associated with sludge formation. Biliary colic is treated with non steroidal analgesic drugs, analgesics and/or spasmolytic agents. PMID- 12649991 TI - [Gallstones--surgical aspects]. AB - Between 10% to 15% of the adult population develop gallstones. Therefore, cholecystectomy is among the most common operations in general surgery. The diagnosis of cholelithiasis depends on the patient's history, clinical findings, laboratory tests and ultrasound examination. Once diagnosis of symptomatic gallbladder disease has been confirmed, laparoscopic cholecystectomy is the treatment of choice. Its advantages in comparison with open surgery are decreased morbidity, costs and hospital stay. Open cholecystectomy is still the treatment of choice for complicated gallstone disease (i.e. cancer, Mirizzi's syndrome, severe inflammation) and in high-risk patients. In case of acute cholecystitis, early laparoscopic cholecystectomy is a safe procedure and is associated with the same benefits as for symptomatic disease. PMID- 12649992 TI - [Salivary calculi--diagnosis and therapy]. AB - The etiology for salivary gland stone formation, one of the most common salivary gland problems, is still unknown. In many cases the diagnosis is easy due to obvious clinical features, but for treatment considerations, imaging studies are very often necessary. Sonography or MR sialography have replaced sialography in the evaluation of patients with sialolithiasis. Besides traditional surgical procedures, new conservative treatment modalities like extracorporeal shock wave lithotripsy or interventional sialendoscopy are now available for the management of salivary gland stones. PMID- 12649993 TI - Complementary medicine and challenges for Western medical practice. PMID- 12649994 TI - Natural death in 2003: are we slipping backwards? PMID- 12649995 TI - Epilepsy and trauma. PMID- 12649996 TI - Transfusion-related acute lung injury. PMID- 12649997 TI - Norwalk or Noroviruses. PMID- 12649998 TI - Shakoor v Situ. [2000] 4 All ER 181. PMID- 12649999 TI - Does a doctor have a duty to provide information and advice about complementary and alternative medicine? AB - It is argued that a doctor has a duty to provide information about reasonably available complementary and alternative medicine treatments where that information would be material to the particular patient or the hypothetical prudent patient. Given the vast array of such treatments available, doctors will want to rely on evidence-based medicine problem-solving skills to ascertain those treatments that are safe and efficacious. While the risk of litigation for failure to provide such information is probably low at this time, given the high rate of patient self-prescribing, it is necessary for a doctor to open a dialogue with a patient about complementary and alternative medicine to address safety concerns. In addition, it is important to facilitate access to the best of conventional and complementary treatments to ensure better health outcomes for the patient. PMID- 12650000 TI - Chinese dragon or toothless tiger? Regulating the professional competence of traditional Chinese medicine practitioners. AB - The escalation in popularity of complementary and alternative medicine (CAM) has begun to stimulate regulatory responses to ensure the safety and efficacy of different modalities. The Therapeutic Goods Authority in Australia oversees a scheme of listing and registration, said to lead the world. Established CAM courses now confer recognised bachelor degrees. Victoria has recently regulated Traditional Chinese Medicine (TCM) practitioners, through the Chinese Medicine Registration Act 2000 (Vic), modelled on legislation regulating medical practitioners. CAM is being integrated into conventional medical (especially general) practice, and calls for the "mainstreaming" of CAM are increasing. Integrating CAM, however, involves a critical incoherence, well illustrated by the Victorian legislation. Clinical competence can only be properly assessed against standards established through scientific validation. If CAM systems, which purport to offer alternatives to science-based medicine, are regulated through conventional instruments, they may well be relinquishing the very identities which set them apart. PMID- 12650001 TI - Obligation to advise of options for treatment--medical doctors and complementary and alternative medicine practitioners. AB - An important aspect of health professional's duty of care is to advise patients of the available options of treatment so that the patient can choose the form of treatment that suits her or his requirements. As CAM becomes more evidence-based and accepted, medical doctors need to consider the extent to which they should provide patients with information about those types of treatments. If a CAM treatment option is evidence-based, there is a strong argument that medical doctors should advise of this option for treatment to satisfy their duty. CAM practitioners should also provide details of options for treatment within their own modality but are not obliged to advise of medical options. PMID- 12650002 TI - General practitioners' perspectives regarding competence and confidentiality in an adolescent with suspected anorexia nervosa: legal and ethical considerations. AB - In Victoria, Australia, the legal position regarding young people's competence to make medical treatment decisions has not been clarified in legislation, and a number of often vague common law decisions must be relied on for guidance. This situation produces a degree of uncertainty about appropriate professional practice, while also potentially impeding young people's rights claims in health care settings. With this in mind, the present research explored general practitioners' competence and confidentiality decisions regarding a 17-year-old female who presented with symptoms of an eating disorder. Questionnaires were sent to a random sample of 500 Victorian general practitioners, of whom 190 responded. After reading a case vignette, general practitioners indicated whether they would find the hypothetical patient competent and if they would maintain her confidentiality. Seventy-three per cent of respondents found the patient competent and most would have maintained confidentiality, at least initially. However, subsequent analysis of the rationales supplied for these decisions revealed a wide diversity in general practitioners' understandings and implementations of extant legal authority. This research highlights the need for general practitioners to be exposed to up-to-date and clinically relevant explanations of contemporary legal positions. PMID- 12650003 TI - Tensions in private health insurance regulation. AB - This article provides an analysis of the regulatory framework of Australian private health insurance linked to four major implicit regulatory objectives: promoting access to health insurance for consumers; promoting financial solvency and industry viability of registered health benefits organisations; promoting competition between registered health benefits organisations; and promoting accountability to consumers. Through an analysis of regulatory changes, case law and policy documents on the performance of the health insurance industry, it is argued that existing health insurance regulation exhibits inevitable tensions due to shifting and often conflicting government objectives about the role of private health insurance. PMID- 12650004 TI - Regulation of online pharmacy: an Australian perspective. AB - This article argues that existing Australian regulations do not adequately cover online pharmacies or Internet advertising of medicines and that existing penalties and sanctions are often ineffective, potentially placing public health and safety at risk. Suggestions are made for future regulatory approaches. It is concluded that as well as an effective program of public education, cautious domestic legislative reform is necessary to ensure specific regulation of Australian online pharmacy practice and Internet advertising of medicines. In addition, the global nature of the Internet demands international co-operation and increased regulator and consumer vigilance. PMID- 12650005 TI - A hospital's non-delegable duty of care. AB - Visiting, honorary and staff medical practitioners, to name but a few, provide medical treatment and services to a variety of "patients", including private, public, in-patients and out-patients. The legal implications arising from the often complex fact situations created by the interactions of these participants and the relationship between hospitals and these participants can lead to hospitals both incurring and avoiding liability for injuries sustained by patients from negligent medical treatment. This article discusses the legal principles governing hospitals' liabilities in this context on the more onerous non-delegable duty of care ground. PMID- 12650006 TI - Facility profile. Modern technology, age-old culture. PMID- 12650007 TI - Building a green future with environmentally friendly construction materials. PMID- 12650008 TI - Penny wise? Financial pressures force a short-term mind-set in capital spending. PMID- 12650009 TI - Powering up! Advice on maintaining your 'on-site electrical utility'. PMID- 12650010 TI - Load'em up. Manufacturers tout automated guided vehicles for department operations. PMID- 12650011 TI - On the air. Now is the time to register your wireless medical telemetry. PMID- 12650012 TI - Special deliveries. Common sense advice on sourcing cleaning supplies. PMID- 12650013 TI - [Are there too many antireflux operations done in Finland?]. PMID- 12650014 TI - [Is cardioversion always needed in atrial fibrillation?]. PMID- 12650015 TI - [Myocyte sarcolemmal and cytoskeleton proteins in inherited muscular diseases]. PMID- 12650016 TI - [Surgical treatment of reflux esophagitis in the Mikkeli Central Hospital during 1997-2001]. PMID- 12650017 TI - [Urination problems]. PMID- 12650018 TI - [Sexuality in medical education]. PMID- 12650019 TI - [Sexuality and the life cycle]. PMID- 12650020 TI - [Female sexual dysfunction]. PMID- 12650021 TI - [Cardiovascular diseases, diabetes and sexuality]. PMID- 12650022 TI - [Cancer and sexuality]. PMID- 12650023 TI - [Sexual dysfunction in neurological disorders]. PMID- 12650024 TI - [Sexual dysfunction in mental disorders]. PMID- 12650025 TI - [What is sexual therapy?]. PMID- 12650026 TI - [Sex instruments as a part of the treatment of a disease]. PMID- 12650027 TI - [Erotic feelings in the physician-patient relationship]. PMID- 12650028 TI - [Diagnosis and therapy of superficial tumors of the urinary bladder]. AB - Among patients with bladder carcinoma, 70-80% present with superficial bladder tumors. This group is composed of papillary tumors Ta, T1 and of flat nonpapillary high-grade carcinoma in situ (Tis). Typical features of superficial tumors are their multifocal origin and high recurrence-rate after initial therapy. The first and most important step in diagnosis and treatment of the bladder cancer is urethrocystoscopy under general or spinal anesthesia with resection of papillary lesions and biopsy of suspicious areas followed by pathologic examination of the surgical specimen. This approach enables total resection of all visible tumors, classification of the tumor and determination of prognostic factors. The aims of intravesical therapy are treatment of Tis and prevention of recurrence and progression after TUR. The most often used agents for intravesical chemotherapy are mitomycin C and adriamycin. This treatment modality prolongs tumor-free interval and reduces early recurrence-rate in about 14% of patients, but cannot reduce late recurrence-rate and progression-rate. BCG intravestical immunotherapy is with 73-87% response rate the most effective treatment modality in Tis. It is also more effective than chemotherapy in prevention of recurrence, but is connected with more frequent side effects. Radical cystectomy is considered in patients who do not respond to local therapy. PMID- 12650029 TI - [IgA nephropathy. Significance of immunoglobulin A glycosylation in pathogenesis and clinical presentation]. AB - Human immunoglobulin A is represented by two structurally and functionally distinct subclasses: IgA1 and IgA2. IgA1, which is almost exclusively present in the mesangial deposits in IgA nephropathy patients, contains in its hinge region three to five O-lined carbohydrate chains. A fraction of IgA1 molecules in the circulation of IgA nephropathy patients exhibits aberrant glycosylation. As a result of changes in glycosylation, the neoepitopes represented by glycans are exposed and recognized by naturally occurring antibodies with antiglycan specifciities, and immune complexes are generated. The deposits of these immune complexes in the glomerular mesangia elicit inflammatory response known as IgA nephropathy. Epidemiological studies have shown that dominant hematuria, either isolated or combined with mild proteinuria, is the most frequent urinary syndrome in glomerulonephritis. The morphologic finding of this syndrome is most frequently IgA nephropathy. Originally considered a benign disease, IgA nephropathy is now recognized as a frequent cause of chronic renal failure. The progression is signalized by increasing proteinuria and hypertension. Therefore, a control of blood pressure and lowering of proteinuria remain the corner-stones of the treatment. Angiotensin converting enzyme inhibitors and AT1 blockers may lower both blood pressure and proteinuria and are now increasingly promoted even for treatment of normotensive patients. Steroids are administered to patients with severe proteinuria. High-doses of fish oil seem to slow down the rate of renal failure. PMID- 12650030 TI - [Clinical importance of semi-quantitative monitoring of lymphomas using the comparative polymerase chain reaction]. AB - BACKGROUND: PCR techniques detecting interchromosomal translocation and clonal immunoglobulin gene rearrangement (IgH) as disease markers in non-Hodgkin's lymphomas (NHL) has been utilised past ten years. However, qualitative PCR detection of persisted minimal residual disease cannot provide clinically useful prognostic information and presently, quantitative approaches are required to predict patient outcome and assess response to the treatment. In some cases, "end point" quantifying techniques, such as comparative PCR, are applicable and the relative estimation of differences in target quantity may serve in disease monitoring rather than absolute number of target copies. METHODS AND RESULTS: Our method of comparative PCR employs co-amplification of sequences of interest (clonal CDR3, bcl2/Jh) and the segment of Hras 1 gene(ras) as an internal standard. Serial dilutions of stored diagnostic DNAs from blood and bone marrow are examined in the same PCR and, after gel densitometry, the amount of initial target is assessed by comparing exponential products of co-amplification. The comparative PCR assay was utilized in monitoring of NHL patients cured either with conventional therapy, or with high-dose regimens and transplantation with stem cells, or with chimaeric anti-CD20 monoclonal antibody (Rituximab). Results from 50 monitored intervals obtained during several months up to several years were supplemented with clinical statements retrospectively. Some of patients became PCR-negative, reappearance of PCR-positivity was observed as well. The decrease or increase of disease marker corresponded to clinical observations. Results obtained from bone marrow were in agreement with those obtained from blood. CONCLUSIONS: End-point quantifying PCR comparative assay may provide an information on the increased risk of relapse and impact of the therapy. The predictive value of these methods depends on the frequency of sample taking and on the sensitivity of the method, which should be monitored in negative cases. PMID- 12650031 TI - [Hypergastrinemia associated with Helicobacter pylori infection and sideropenic anemia in a 15-year-old girl]. AB - H. pylori is a major cause of primary chronic gastritis and peptic ulcer disease in children. The authors give an account of H. pylori infection (cagA+, vacA+) in a 15-year-old girl where the initial clinical features included fatigue, collapses, and anorexia, elevated serum gastrin level (> 1000 mIU/l) raised the suspicion of gastrinoma. H. pylori gastric infection was also associated with iron-deficiency anemia. After treatment for H. pylori infection (omeprazole, clarithromycin, amoxycillin), clinical symptoms improved consistently, the serum gastrin level was repeteadly quite normal and hematologic and iron profiles were within the normal range. There is compelling evidence that H. pylori must be taken into account as a cause of hypergastrinemia other than gastrinoma in childhood. PMID- 12650032 TI - [The age of formation of Czech clinical medicine]. AB - Era of Emperor Francis Joseph I is said to be a golden age for the Czech nation. It can be found in numerous panegyric articles to any jubilee of the emperor's rule. What was formally dictated by respect brought by education and by the system of the Greek and Roman tradition adopted in Austria in Middle Ages, seams to be valid today as most of the contemporary technical and economical progress roots within those days. The Czech cultures namely music and art reached international acknowledgement. Though with difficulties, Czech achieved in education and in science as a full-fledged language. After the year 1848 an average citizen was entitled to such freedom as never before. Technical, economical and cultural progress enabled real ascent of the Czech society and its social differentiation. In sixties, after the Austria-Hungary Alignment, Hapsburg government undertook no serious restrains. Such development was nothing unusual. Similar one underwent after the period of storms all European societies from south to north and form west to east. They brought ideas of the French revolution and years 1848/1849 are therefore called "the spring of European nations". In all countries where revolutionary ideas were represented and various countermeasures were accepted, governments were forced to accept temporary arrangements (in Austrian monarchy it was the promise of constitution, language compromise etc.). Nevertheless, in the second half of the 19th century the most important condition for further revival was the long period of peace and stability of international relations. The internal stability of the Austrian monarchy was achieved for long time by the Austria-Hungary Alignment in 1867. After the lost battle at Hradec Kralove in summer 1866 it became clear that contemporary centralistic organization of the state, balancing between absolutism and constitutionalism is not further tenable. Years long pressure of patriotic forces in the parliament brought about division of the Charles-Ferdinand University. Important role in the partition of the Czech Medical Faculty from the German one had the foundation of the Czech Medical Society and the Czech Medical Journal in 1862 patronaged by professors Purkyne, Eiselt, Gregr and others. That was the platform for the conception of the Czech medical terminology. Partition of faculties was decided by the law, which was accepted at February 28, 1882. The Czech Medical Faculty was opened only at 1883 because Emperor resolved that professors themselves had to decide to either faculty their clinics would belong. Only professor Eiselt with the First clinic of internal medicine, Weiss with the clinic of surgery and Streng with clinic of obstetrics were assigned to the Czech faculty. Till the official opening on October 15, 1882 all other departments and clinics had to be organized. Though Professor Vilem Weiss as the head of the department and dean of the faculty had his opening Czech lecture already on April 28, 1882, due to material and personal reasons the Czech Medical Faculty was opened only one year later than Faculties of law and philosophy. Most of the eminent members of the Medical faculty remained at the German faculty. However, having higher number of students the Czech faculty became larger and was attended by students from the whole Slavonic world. PMID- 12650033 TI - [Value of spiral CT and MRI (1.5 T) in preoperative diagnosis of tumors of the head of the pancreas]. AB - OBJECTIVE: Prospective comparison study of helical computed tomography (HCT) and magnetic resonance imaging (MRI, 1.5-tesla field strength) with regard to their role in planning surgical therapy in a defined, homogeneous patient collective with tumors of the pancreatic head. PATIENTS AND METHODS: Sixty-one patients with clinical/ultrasound evidence of tumor were examined by HCT and MRI. Findings were compared with surgical anatomy and histology using a data documentation sheet. Factors evaluated included the malignancy of the tumors and the probability of correct diagnosis in relation to tumor size, as well as the occurrence of lymph node and distant metastases, arterial and portal-venous infiltration, extrapancreatic infiltration, determination of resectability and tumor staging (TNM classification). These factors were compared on the basis of their respective sensitivity, specificity, negative and positive predictive value and accuracy. RESULTS: Malignancy was correctly diagnosed by HCT and MRI in 82% and 77%, respectively, with a corresponding sensitivity for malignancy of 86% and 68%. Tumors with diameters > 2 cm were detected in 100% of cases by HCT and in 88% by MRI, while tumors with diameters < 2 cm were correctly diagnosed by HCT and MRI in 60% and 40% of cases. Respective rates of correct diagnosis by HCT and MRI were 80% and 66% for extrapancreatic infiltration; 76% and 79% for pancreatic cysts; 87% and 87% for calcifications; and 77% and 92% for atrophy. The diagnostic accuracy for vascular infiltration by HCT and MRI stood at 59% and 77% with sensitivities of 88% and 75%, respectively. Local resectability was correctly determined by HCT in 71%, by MRI in 62% of cases. Lymph node metastases were correctly identified by HCT and MRI with an accuracy of 76% and 72%, while distant metastases were correctly diagnosed by HCT and MRI with an accuracy of 93% and 93%. Correct T stage was identified by HCT in 66%, by MRI in 60%. HCT tended to assign patients to a higher T stage than later determined by surgery and/or histology. CONCLUSION: Our data do not indicate a clear superiority for either HCT or MRI in the diagnosis of pancreatic malignancies. PMID- 12650034 TI - [Pre- and post-interventional follow-up of stent graphs with single and multi slice spiral computerized tomography]. AB - PURPOSE: Could contrast enhanced computed tomography replace conventional catheter angiography in the preoperative evaluation of aortic aneurysms before stentgraft implantation? Does multi slice computed tomography have advantages comparing to single slice computed tomography? METHODS: We compared 15 single slice with 13 multi slice computed tomography studies and reconstructions according to there ability to replace conventional catheter angiography in the preoperative evaluation. In the postoperative follow-up 60 single slice computed tomography studies were compared to 60 multi slice studies. RESULTS: By using appropriate reconstructions the spiral computed tomography studies could deliver all information necessary for implantation. In our study no advantage of multi slice computed tomography could be detected. In the postoperative evaluation of stentgrafts our computed tomography scan protocol could detect all relevant complications in single and multi slice computed tomography. No significant advantage of multi slice computed tomography could be detected. CONCLUSION: Contrast enhanced computed tomography is equal to conventional catheter angiography in the pre- and postoperative evaluation of stengrafts. No significant advantage of multi slice versus single slice could be detected. PMID- 12650035 TI - [CO2 angiography of transplanted kidneys]. AB - PURPOSE: To compare image quality and diagnostic accuracy of carbon dioxide (CO2) and iodinated contrast medium as a contast medium in renal transplant artery angiography. MATERIAL AND METHODS: In this prospective, non-randomized, intra individual study, we examined 17 patients. Digital subtraction angiography (DSA) was performed first with CO2 to find the optimal projection. Then at least one confirming run was performed with CM ("gold standard"). The quality of the angiographic studies with CO2 and CM and their diagnostic accuracy were compared. Blood creatinine levels were monitored during three days after angiography. RESULTS: Three of four renal transplant artery stenoses were diagnosed correctly with CO2. The false-negative CO2 angiography was due to poor contrast. There were no false-positive results with CO2. Regardless the inferior image quality of CO2 angiography, its positive predictive value was 100%. Renal function was not compromised. CONCLUSION: The primary use of CO2 as a contrast medium for the angiographic evaluation of renal transplant arteries is feasible and practical. It reduces the amount of CM needed. However, in order to confirm the diagnosis, at least one additional series with CM should be performed. PMID- 12650036 TI - [Regional lung edema in acute mitral valve insufficiency following chordae tendineae rupture with prolapse of the posterior mitral valve leaflet]. AB - An unilateral or predominantly lobar pulmonary edema is an unusual clinical or radiological finding, often misdiagnosed as one of the more common causes of focal lung disease. We report 2 cases of a regional pulmonary edema caused by the acute onset of a severe mitral insufficiency after the rupture of chordae tendinae resulting in a prolaps of the posterior mitral leaflet. In both cases the regional pulmonary edema was initially misdiagnosed as a pneumonic infiltration, which delayed the cardiological diagnostical procedures and the surgical intervention. The mechanism of the regional edema is an excentric recurgitation jet into the left atrium, which is usually directed to the orifice of the right upper lobe pulmonary vein which increases the hydrostatic vascular pressure in the corresponding lung segment. For the confirmation of the diagnosis, transesophageal echogradiographic is helpful in documenting the direction of the regurgitant flow and detecting differential gradients between the right and left pulmonary venous systems. The pulmonary infiltrations, which persisted for several weeks, disappeared within a few days after surgical mitral valve-reconstruction in both cases. PMID- 12650037 TI - [Clinical symptoms and diagnostic imaging in moyamoya disease]. AB - OBJECTIVE: Presentation of typical imaging findings and diagnostic approaches in patients suspected of moyamoya disease. MATERIAL AND METHODS: Two female patients (24 resp. 44 years old) presenting with equivocal neurological symptoms (headache, recurrent monoparesis, choreiformic ataxia, grand mal seizure) were examined by means of colour-coded ultrasound, selective catheter-based angiography (DSA), CT and MRI including MR angiography. RESULTS: Cranial CT and MRI showed normal findings in one patient, and focal cerebral infarctions of different age in the other. In both cases, colour-coded ultrasound was indicative in detecting occlusive disease of the internal carotid arteries. Both, MR angiography as well as catheter-based angiography revealed the pathology of the culprid vessel, in addition catheter-based angiography was more accurate in depicting moyamoya collaterals at the skull base. CONCLUSION: In moyamoya disease colour-coded ultrasound is diagnostic for the lesion of the internal carotid artery, MRI resp. MRA depict the vascular and parenchymal lesion, whereas catheter-based DSA by providing information about the main vessel and collateral system allows the staging of the disease. PMID- 12650038 TI - Electronic archiving with PubMed Central--next steps into the future. PMID- 12650039 TI - An ethicist's commentary on veterinary "nurse practitioners". PMID- 12650041 TI - Medullary trichomalacia in 6 German shepherd dogs. AB - Medullary trichomalacia is the name proposed for a hair shaft abnormality that was recognized in 6 German shepherd dogs. Affected dogs had multifocal areas of broken hairs, especially on the dorsolateral trunk. Microscopic examination of hair shafts revealed focal areas of loss of architecture, swelling, and apparent softening of the medulla, followed by longitudinal (length-wise) splitting and breakage of the hair shaft. No cause could be found. Affected dogs were otherwise healthy, and apparent spontaneous recovery was the usual outcome. Relapses may occur. PMID- 12650040 TI - The effects of 3 gonadorelin products on luteinizing hormone release, ovulation, and follicular wave emergence in cattle. AB - The objective was to determine luteinizing hormone (LH) secretion and follicular dynamics in cattle following administration of 3 gonadorelin formulations that are commercially available in Canada. In experiment 1, nonlactating Holstein cows (n = 4 per group) were randomly assigned to receive 100 micrograms gonadorelin diacetate tetrahydrate, intramuscularly (C; Cystorelin, or FE; Fertagyl). Blood samples (for LH analysis) were collected 0, 1, 2, and 4 hours after treatment. In experiment 2, nonlactating Holstein cows (n = 10 per group) were randomly allocated to receive 100 micrograms gonadorelin, intramuscularly as follows: 2 mL of C; 1 mL of FE; or 2 mL of Factrel (FA, gonadorelin hydrochloride). Gonadorelin treatment was done on days 6 or 7 after ovulation and blood samples for LH analysis were collected at 0, 1, 2, 4, and 6 hours after treatment. Ovaries were examined by ultrasonography, twice daily, to detect ovulation. A replicate was conducted using only C (n = 10) or FE (n = 10); blood samples were collected at 0, 1, 2, 3, and 4 hours. In experiment 3, beef heifers (n = 10 per group) were randomly assigned to receive 1 of 3 GnRH gonadorelin treatments (as in the first phase of experiment 2) on days 6 or 7 after ovulation and blood samples were collected at 0, 0.5, 1, 1.5, 2, and 4 hours. In experiments 2 and 3, both mean and mean peak plasma LH concentrations were higher (P < 0.05) in cattle treated with C. The proportion of dominant follicles that ovulated was higher (P < 0.02) in Holstein cows treated with C than in those treated with FE or FA (18/19, 11/19, and 4/7, respectively), but there was no significant difference among the products in beef heifers (6/10, 6/10, and 4/10, respectively). No significant differences were found in the interval from treatment to the emergence of the next follicular wave. In summary, C induced a greater LH release and this resulted in a higher ovulatory rate in Holstein cows but not in beef heifers. PMID- 12650042 TI - Intradural vasculitis and hemorrhage in full sibling Welsh springer spaniels. AB - Two, full sibling, Welsh springer spaniel presented at 8 and 18 mo of age with rapidly progressive ataxia, recumbency, and pyrexia. The spinal cord contained extensive subdural hemorrhage and, in 1 dog, suppurative and necrotizing arteritis in the dura. The findings suggest a familial form of canine juvenile polyarteritis syndrome. PMID- 12650043 TI - Hip dysplasia in a 6-year-old Salz ram. AB - A 6-year old Salz ram was presented with a history of poor body condition, progressive gait abnormalities, pelvic limb lameness, and difficulty with copulation. Based on the history, clinical signs, hip palpation, and radiography, a diagnosis of hip dysplasia, previously unreported in sheep, was made. PMID- 12650044 TI - Acetaminophen toxicosis in a Dalmatian. AB - An 11-year-old, spayed female Dalmatian was presented with suspected acetaminophen toxicosis. The dog was severely depressed. Methemoglobinemia, facial edema, and hemoglobinuria responded to treatment with intravenous fluids, N-acetylcysteine, ascorbic acid, and sodium bicarbonate. There was no clinical evidence of hepatic damage typical of acetaminophen toxicity in the dog. PMID- 12650045 TI - Unilateral white line disease and laminitis in a quarter horse mare. AB - A 5-year-old quarter horse mare presented with unilateral, severe, chronic forelimb lameness. Radiographs revealed extensive hoof wall separation and capsular rotation of the distal phalanx. Treatment included dorsal hoof wall resection, phenylbutazone, a bar shoe, and stall rest. Whether white line disease or laminitis was the primary lesion remains unclear. PMID- 12650046 TI - Hypertensive retinopathy in a cat. AB - A 12-year-old cat presented for sudden blindness was diagnosed with hypertensive retinopathy on the basis of ophthalmologic and ultrasonic examination. Renal failure due to a large intranephric cyst obstructing the right ureter and renal artery was the suggested cause of the systemic hypertension. The cat died 8 hours after unilateral nephrectomy. PMID- 12650047 TI - The Ordre des medecins veterinaires du Quebec celebrates its centennial. PMID- 12650048 TI - What's new in orthopedics? Part 2. PMID- 12650049 TI - Changing protocols surrounding euthanasia. PMID- 12650050 TI - Coccidiomycosis in a dog. PMID- 12650051 TI - [Methicillin-resistant Staphylococcus aureus (MRSA) with high resistance to mupirocin in hospitals of the Gdansk region]. AB - Occurrence of high-level mupirocin resistance in methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from 18 hospitals in Gdansk area was determined. The study was carried out on 190 MRSA isolated in 1997-2000 from various clinical samples. The strains were tested for high-level mupirocin resistance by 200 micrograms mupirocin disc. The minimum inhibitory concentrations (MIC) for methicillin were estimated by agar dilution. Sensitivity to other antibiotics was determined in disc-diffusion method and to vancomycin in agar dilution method additionally. The strains were typed by set of 10 experimental phages and compared by the method of PCR-RFLP analysis of coagulase gen restriction fragment length polymorphism. There were low frequency of high level mupirocin resistance in MRSA strains (4.7%) that were found only in 3 hospitals, in 6 patients. All of them were high-resistant also to methicillin and resistant to doxycyclin, gentamycin, erytromycin, klindamycin, ciprofloksacin, rifampicin, resistant or intermediate sensitive to fusidic acid but sensitive to vancomycin, teikoplanin and bacitracin. The origin all of the MRSA strains high resistant to mupirocin probably was the same, except one strain, because they were belonged to one genetic type and possessed the same phage pattern. PMID- 12650052 TI - [Effect of subinhibitory levels of selected antibiotics on susceptibility of Staphylococcus aureus strains to phagocytosis and killing by rabbit granulocytes]. AB - The aim of the study was to assess the of chosen antibiotics in subinhibitory concentrations (sub-MIC) on the sensitive of Staphylococcus aureus cells to phagocytosis and killing by rabbit granulocytes. The following antibiotics were used: cloxacillin, cefadroxil, cefuroxim, cefotaxim, gentamicin, netilmicin, lincomicin, doxycycline and riphamicin. A total of 144 S. aureus strains with varied sensitivity to these antibiotics were selected for the study. The experiment used granulocytes isolated from rabbit blood and S. aureus strains incubated for 18 h in TSB broth containing antibiotics in the concentrations of 0.1 MIC, 0.2 MIC and 0.5 MIC, and in the antibiotics-free medium. Phagocytosis was assessed by the method of differential staining with acridine orange and crystal violet, allowing simultaneous determination of phagocytised and killed S. aureus cell counts. The findings revealed that the culture of S. aureus in the presence of all the antibiotics used in subinhibitory concentrations increased significantly the susceptibility of most S. aureus strains to phagocytosis and killing by granulocytes. The above effect usually occurred in the concentrations of 0.1 MIC (54.2%), more seldom in 0.2 MIC (13%) and 0.5 MIC (15% of strains). Each group of S. aureus contained some which showed no change in susceptibility following culture with the chemotherapeutic agents in subinhibitory concentrations (26.3%). Insensitive strains to the subinhibitory effects were equally common among susceptible (27%), intermediate (23%) and resistant (26%) strains of S. aureus to the antibiotics used. No statistically significant reduction was noted in phagocytosis or killing by rabbit granulocytes. No correlation was observed between the susceptibility to the subinhibitory effects of the antibiotics involved and their biochemical mechanisms. PMID- 12650053 TI - [Drug sensitivity of Enterococcus sp. strains isolated from clinical material or from healthy persons]. AB - The aim of this study was to evaluate the drug susceptibility of Enterococcus sp. strains isolated in 2000-2001, from patients of five Warsaw's hospitals (154 strains) and from fecal samples of healthy persons (33 strains). On biochemical reaction profiles species of clinical enterococci were identified as: E. faecalis -66.2%, E. faecium--29.2%, E. hirae--1.9%, E. gallinarum--1.3%, E. casseliflavus- 0.6% and E. avium--0.6%. The species of enterococci from stool's samples were identified as: E. faecalis--28 strains, E. durans--2 strains and single strains: E. faecium, E. gallinarum and E. casseliflavus. Susceptibility to 20 antibiotics was tested by disc diffusion method. None of these 187 enterococcal strains was vancomycin resistant; 3 strains of E. gallinarum and 1--E. casseliflavus demonstrated intermediately susceptibility to vancomycin, but they were susceptible to teicoplanin--phenotype Van C. Among clinical strains were resistant to penicillin--33.3% of E. faecalis and 100% of E. faecium, to ampicillin--over 80% of E. faecium and 1 strain of E. faecalis. None of these strains produced beta-lactamase. High-level resistance to aminoglicoside was expressed by 48 strains (47.1%) E. faecalis and 36 (80%) E. faecium isolated from clinical specimens. Both--HLR to streptomycin and gentamycin were found in 28.3% of E. faecalis and 68.9% of E. faecium. Among 33 strains isolated from fecal samples of healthy persons--3 of E. faecalis were resistant to streptomycin and one was resistant to both gentamicin and streptomycin. In general, enterococcal strains isolated from samples of healthy persons were susceptible to the most of used antibiotics. But to rifampicin none of these strains were susceptible. There were about 40% of E. faecalis strains isolated from healthy persons, resistant to tetracyline. PMID- 12650054 TI - [Susceptibility of enterococci to natural and synthetic iron chelators]. AB - A total of 79 strains of enterococci belonging to 10 species were tested for susceptibility to natural and synthetic iron chelators. All strains produced siderophores. These enterococci were susceptible to three synthetic iron chelators only: 8-hydroxyquinoline, disodium versenate (EDTA) and o phenanthroline. They were resistant to all other synthetic chelators: ethylenediamine-di(o-hydroxyphenylacetic acid) (EDDHA), nitrilotriacetate, 2,2' bipiridyl, salicylic acid, 8-hydroxy-5-sulphonic acid and to all natural chelators: ovotransferrine, human apotransferrine, horse apoferritine, desferrioxamine B, ferrichrome and rhodotorulic acid. The relations between susceptibility/resistance, iron assimilation and structure and stability constants of iron chelators were discussed. PMID- 12650055 TI - [Phage types and plasmid profiles of plasmid DNA strains of Salmonella enterica subsp. enterica ser. Enteritidis (S. Enteritidis) isolated from food poisoning outbreaks in 2001]. AB - Salmonella Enteritidis strains are the most often isolated Salmonella serovar in Poland. In the present study, phage typing, antibiotic resistance testing and plasmid profile analysis, have been applied to characterise 41 Polish S. Enteritidis isolates originated from human cases of salmonellosis and from other sources. The typing phages of Ward and colleagues scheme were used to type a total of 41 S. Enteritidis strains coming from Poland. All 41 strains were typable and 5 different phage types were observed. Among 41 strains tested, both PT6 and PT21 were recognized in the 15 strains (36.6%). Nine strains (22%) belonged to phage type 8. The others PTs were represented by small amount of strains (PT1var and PT4). Among all tested isolates only 4 different plasmid profiles were observed. Of the 41 strains investigated, 16 (39%) contained the 57 kb plasmid alone. The remaining 25 strains (61%) except 57 kb plasmid, possessed additional DNA particles. The probable phage type conversion of PT21 to PT1var strain, possibly connected with smaller DNA particle presence, was observed. This hypothesis needs confirmation. The real S. Enteritidis epidemiological situation in Poland should be known after introducing of systematic, annual research program. PMID- 12650056 TI - [Investigation of hydrophobicity of Proteus vulgaris strains and ability of Proteus vulgaris and Proteus penneri strains to penetrate bladder membrane HCV T 29 cells ]. AB - Proteus bacilli play a particularly important role in urinary tract infections (UTI). Fimbriae and adherence ability and hemolysins production (HpmA, HlyA) are one of the factors of pathogenicity of these bacteria. In this paper we describe the invasion of HCV T-29 transitional bladder urothelial cells carcinoma strains of P. penneri, as well as P. vulgaris strains belonging to different serogroups. The cytotoxic effect was observed at 8 hour of incubation of the tested cells with P. vulgaris O21 and the same effect (complete lysis) at 6 hours by P. vulgaris O4 (this strain manifests maximal activity in the production of HlyA hemolysin). P. penneri strains, produce different types of fimbriae, expressed similar bacterial invasiveness. The hydrophobic properties of 25 P. vulgaris strains were also tested and only 3 strains occur to have hydrophobic cell surface. PMID- 12650057 TI - [Evaluation of usefulness for selected virulence markers for identifying pathogenic Yersinia enterocolitica strains. IV. Genes myfA and ureC]. AB - We check by polymerase chain reaction (PCR) the presence of gene ureC and myfA, encoding subunits of urease and Myf fimbriae, among clinical and food-originated strains of Yersinia to determine their usefulness as molecular virulence markers of Y. enterocolitica. The examinations were done on 130 clinical strains of Y. enterocolitica O:3/4 isolated in Poland from humans. All strains were obtained from stool and possessed the virulence plasmid pYV. In addition 40 isogenic, plasmid-cured strains were tested. The 52 strains including Y. enterocolitica (biotype 1A, 4, 2 and 1B), Y. pseudotuberculosis, Y. intermedia, Y. frederiksenii, Y. kristensenii, E. coli, Citrobatcer, Shigella and Salmonella were used as controls. The PCR assay resulted in detection of genes: ureC and myfA in genomic DNA of all 130 tested clinical strains of Y. enterocolitica pYV+, as well as in plasmid cured strains. Furthermore, ureC was found in all tested strains of Y. enterocolitica biotype A1 and in one strain of Y. intermedia and Y. kristensenii. In contrast to ureC, myfA was detected only in strains of Y. enterocolitica considered as pathogenic. Obtained results show, gene myfA seems to be the reliable virulence marker of Y. enterocolitica, whereas ureC is not recommended for identification of pathogenic strains of this species. PMID- 12650058 TI - [Prevalence of enterotoxigenic Bacteroides fragilis strains (ETBF) in the gut of children with clinical diagnosis of antibiotic associated diarrhoea (AAD)]. AB - Prevalence of enterotoxin producing Bacteroides fragilis (ETBF) strains in faecal samples of children with clinical diagnosis antibiotic associated diarrhoea (AAD) was investigated. Out of faecal samples collected from sixty children, thirty C. difficile strains were isolated. Enterotoxigenic B. fragilis (ETBF) strains were cultured from four children what made 6.7% of investigated faecal samples. Out of two samples toxinogenic C. difficile strains [tox A(+) tox B(+)] were cultured together with enterotoxinogenic B. fragilis. From sample of one child C. difficile A negative/B positive strains [tox A(-) tox B(+)] was found together with B. fragilis (ETBF). From faecal sample of one child enterotoxinogenic B. fragilis only was isolated. It was shown that in the gut of children with clinical diagnosis of (AAD) enterotoxinogenic B. fragilis (ETBF) can be present. B. fragilis (ETBF) can be observed in concomitance with toxinogenic C. difficile. PMID- 12650059 TI - [Use of reactions with Limulus amoebocyte lysate (LAL) to determine biological activity of lipopolysaccharides from reference and clinical strains of the Bacteroides fragilis group]. AB - The aim of this study was to determine and compare a biological activity of lipopolysaccharides (LPS) from reference and clinical strains of strictly anaerobic bacteria belonging to the Bacteroides fragilis group (BFG) by means of quantitative, photometric BET (LAL) method with Limulus polyphemus amoebocyte lysate and chromogenic substrate S-2423. Lipopolysaccharides of five BFG species were extracted by Westphal and Jann method (1965) from eight reference and two clinical strains of B. fragilis group. Crude LPS preparations were purified according to the procedure described by Gmeiner (1975) with ultracentrifugation and nuclease treatment. Biological activities of bacterial endotoxins were determined by quantitative BET method with chromogenic substrate S-2423 (ENDOCHROME kit, Charles River Endosafe Ltd., USA). Tests were performed according to the producer's recommendations. E. coli O55:B5 LPS was applied to compare its activity in reaction with LAL reagent with activities of LPS preparations from rods of the Bacteroides genus. Among examined bacterial compounds the most active in BET method was E. coli O55:B5 LPS. Activities of lipopolysaccharides from five species of BFG rods in reaction with Limulus amoebocyte lysate were differentiated. Greater ability to activate LAL proenzyme revealed lipopolysaccharides of these species of the Bacteroides genus, which are important from the clinical point of view--B. fragilis and B. thetaiotaomicron. PMID- 12650060 TI - [Cultivation of fungi from fecal specimens in cases of antibiotic associated diarrhea (AAD)]. AB - The aim of performed examinations was to isolate, identify and determine a drug susceptibility of fungi cultured from faecal specimens submitted for detection of Clostridium difficile in cases of antibiotic-associated diarrhoea (AAD). One hundred samples of diarrhoeic faeces were examined using routine bacteriological methods (isolation and identification of C. difficile), serological test (detection of C. difficile toxins A/B) and mycological methods (isolation, identification and drug susceptibility testing of fungi). Out of twenty seven specimens of diarrhoeic faeces fungal strains were isolated, in 20 samples C. difficile strain and/or C. difficile toxins A/B were detected, in 23 specimens fungal strains, C. difficile strains and/or toxins A/B of this species were present. The most active in vitro agent against cultured fungal strains was nystatin. In conclusion it can be stated, that fungal strains are responsible for some cases of antibiotic-associated diarrhoea. So, mycological diagnostics of faecal samples from patients with diarrhoea after antibiotic therapy is necessary. Cases of diarrhoea with mixed bacterial and fungal aetiology (C. difficile + yeast-like fungus) were observed. PMID- 12650061 TI - [Misery in youth and diligent all my life]. PMID- 12650062 TI - [Suppression of neurotransmitter release by presynaptic metabotropic glutamate receptors]. AB - Activation of ionotropic receptors by glutamate mediates most of the fast excitatory synaptic transmission in mammalian central nervous system. In addition, they are involved in excitotoxic neuronal cell death that occurs in a variety of neurological disorders if these receptors are excessively activated. Metabotropic glutamate receptors (mGluRs) are a G-protein coupled receptor family and they are divided into three groups. Both group II and group III of mGluRs are presynaptically localized on the glutamatergic terminals, and provide a negative feedback to modulate the release of glutamate. Recent data also showed that some mGluRs are presented on non-glutamatergic neurons, such as GABAergic terminals, where mGluRs suppress GABA release when they are activated. Further investigation of mGluRs may lead to the development of novel, safe and effective pharmacological agents to treat a range of neurological disorders and neurodegenerative diseases by preventing the excessive glutamate release. PMID- 12650063 TI - [Progress in research on heat shock proteins in cardiovascular system]. AB - Diverse physiological stresses (e.g., heat shock, ischemia and hemodynamics) produce multiple changes in a cell that affect metabolic processes and cellular structures. In response, stress proteins are synthesized, and are thought to mediate cytoprotection through biological function as molecular chaperones. Accumulated evidence indicates that HSP70, HSP90, HSP47, HSP32 and HSP27 play an important role during cardiac ischemia, ischemia preconditioning, cardiac hypertrophy and vascular wall injury. PMID- 12650064 TI - [A review of the research progress on LIM protein family]. AB - LIM Proteins contain one or more LIM domains. Proteins of this family capable of interacting with many types of proteins have been identified including structural protein, kinase, and several classes of transcription factors. These interactions have been discovered to display important roles in a variety of fundamental processes including transcriptional regulation, development of cell types and organization of the cytoskeleton. This article reviews recent advances in the characterization of members of the LIM protein family, their biology roles, LIM domain binding protein and the function in heart development. PMID- 12650065 TI - [Estrogen receptor subtypes and the regulatory effect of receptor ligand binding on gene transcription]. AB - This review introduces the structure, functions, tissue distribution, physiologic roles of estrogen receptor subtypes (ER alpha and ER beta) along with transcriptional activities of estrogen receptor ligands and the mechanism of the modulatory pathway and tissue specific property of selective estrogen receptor modulators (SERMs), and phytoestrogens. This article is to provide a systemic approach for increasing the selectivity of estrogenic drug and optimizing the clinically based strategy of drug design. Differences exist between ER alpha and ER beta in structure, function, tissue distribution, physiologic roles and response to ligands, as well as the regulating effect on gene transcription, which are mainly determined by different co-regulators they recruit respectively. PMID- 12650066 TI - [Neurotrophic factors and neural stem cells]. AB - Neurotrophic factors includes nerve growth factors (NGF) family, glial cell line derived neurotrophic factors (GDNF) family and other neurotrophic factors. NGF, BDNF, NT-3, NT-4/5 and NT-6 constitute NGF family (the neurotrophins), which may enhance both survival and differentiation of neurons from the EGF-responsive hippocampal and the subventricular zone neural stem cells. GDNF family includes GDNF, NTN, PSP and ART, whose major role is in peripheral neurodevelopment. This family enhance proliferation and survival of enteric neural crest precursor cells and is critical for the development of peripheral sensory nerve. Other growth factors such as bFGF and EGF can enhance proliferation and survival of neural stem cell. CNTF and LIF are critical for differentiation of neural stem cells. PMID- 12650067 TI - [Processing of optic flow information during self-motion]. AB - For human beings and animals, the observer encounters the so-called optic flow during locomotion, the image motion pattern of the environment on the retina, which includes three basic modes of motion: rotation, expansion/contraction and translation. The analysis of optic flow information is vital for determining the direction and velocity of locomotion and has been a hotspot of the studies on visual information processing in recent years. In this article, some progresses in psychophysics and electrophysiology studies on optic flow are introduced and the neural mechanism of optic flow processing in mammalian brain is discussed. PMID- 12650068 TI - [The use of gene expression differential display analysis technics in trauma research]. AB - Gene is the regulation core of cell proliferation, differentiation and maturation. It is also the decisive factor of occurrence, progression and prognosis of many diseases. The changes in gene expression will inevitably lead to many kinds of abnormal in cells, tissue, organs even in whole biological organism. Trauma, as well as other stimulators, may bring a series complications through the abnormal expression of genes following injury. With the development of bioinformatics and molecular biology, a series of assays (so called gene expression differential display analysis technics, such as DNA microarray) have been established, which have been used to analyze differentially expressed genes effectively. These methods have been widely used in the research of tumor and other disease. This review will focus to some usage of the ways in traumatology. PMID- 12650069 TI - [Chronic inflammation, autoimmunity and atherosclerosis]. AB - Atherosclerosis is an inflammatory disease. There are many immune cells filled in atherosclerotic lesions, particularly in unstable plaques. The understanding on the recruitment, differentiation and activation of immune cells in atherosclerosis have been expanded in recent years. Epidemiological surveys show many associated viral and bacterial infections. Preliminary studies have identified several autoantigens and raised the "autoimmune hypothesis". Based on this novel viewpoint, the immunomodulation and immunization have been taken as the new strategies for prevention and treatment of atherosclerosis. This will markedly improve our research level and identify new therapeutic targets in atherosclerosis. PMID- 12650070 TI - [The relationship of oxytocin, vasopressin and stress]. PMID- 12650071 TI - [Cellular and molecular mechanism of cardiovascular disease resulting from hyperhomocysteinemia]. PMID- 12650072 TI - [New typical guidance cues and connection development in hippocampus]. PMID- 12650073 TI - [Genetic regulation of circadian clock]. PMID- 12650074 TI - [The bi-directional modulation of endogenous pain modulating system]. PMID- 12650075 TI - [Evolution of the adreno receptor]. PMID- 12650076 TI - [The technique of single-cell reverse transcription polymerase chain reaction and its application]. PMID- 12650077 TI - [Prolactin family and mammal pregnancy]. PMID- 12650078 TI - [Embryonic regulation in human embryo implantation]. PMID- 12650079 TI - [The theoretical system and application of reporter gene technology]. PMID- 12650080 TI - [The choice and research trend of reporter gene]. PMID- 12650081 TI - [Interleukin-1 in brain injury after seizure]. PMID- 12650082 TI - [Effects of neural cell adhesion molecule and sulfated polysaccharides on the processes of neural development, neurite outgrowth, synaptic plasticity, learning and memory]. PMID- 12650083 TI - [Role of O-GlcNAc in signal transduction]. PMID- 12650084 TI - Consumer access to health care information: its effect on the physician-patient relationship. AB - Studies have shown that physicians exchange minimal information with patients during a patient encounter. Yet patients of all socioeconomic backgrounds increasingly desire and seek medical information, especially over the Internet. This change in patient behavior is likely to have a profound effect on the physician-patient relationship. Many physicians react defensively when patients bring them information on their condition with questions about traditional and non-traditional therapies. This can degrade trust and communication, both key elements in the physician-patient relationship. There are several models of the physician-patient relationship that are described. The deliberative or collaborative model, where the physician helps the patient choose their preferred health-related values, represents our societal norm. Such a model is conducive to physicians acting as a directed facilitator for medical information acquisition by patients. It is also consistent with patients' desire to obtain on-line information under the guidance of a physician. Such a model can not only aid patient's acquisition of medical information, but also help patients assess the quality of information, and help them apply the information to their personal situation. The adoption of this new role for physicians is likely to enhance the therapeutic aspects of the physician-patient relationship. PMID- 12650085 TI - Oral health of young Alaska Native children and their caregivers in Southwestern Alaska. AB - OBJECTIVE: We sought to determine the prevalence of caries and associated risk factors in young Native children and their caregivers in two communities in rural Alaska. METHODS: A random sample of Alaska Native children between 12-36 months and a subset of their caregivers from two Southwestern Alaska communities were examined for dental decay. Caregivers completed a 43-item questionnaire about oral hygiene, dietary and other practices. RESULTS: Of the 65 children examined, 59% had evidence of decay. Among the 41 primary caregivers examined, 98% had experienced dental decay with an average of 11.4 decayed, missing, and filled teeth. On linear regression analyses, factors significantly associated with more decay in the children included a child taking a juice-containing bottle to bed, eating candy 1 or more times per day, and higher caregiver oral S. mutans counts. CONCLUSIONS: Our results suggest that preventive efforts for children at high risk for dental decay should begin early in life, should emphasize decreasing candy and bedtime juice bottle use, and should consider the caregivers' oral health status in addition to the child's. PMID- 12650086 TI - Preliminary study on total mercury in the common prepared subsistence foods of a rural Alaskan village. AB - Total mercury (THg), which includes neurotoxic methylmercury (MeHg), poses a health risk to subsistence food users in the circumpolar north. Low levels of THg, usually below 200 ng/g, have been reported in fish muscle from both returning salmon and freshwater species samples on the Kuskokwim River in southwest Alaska. Alaska subsistence users use fish all year long, with both salmon and freshwater species stored for winter usage. A recent study showed low mercury levels in the hair of villagers and suggested that in the last 25 years, MeHg exposure has remained the same or slightly decreased in the Bethel region of Alaska. Although several previous studies have measured THg in hair of Alaskan subsistence food users, few studies have looked at the THg levels in the prepared foods of rural Alaskans. Several types of subsistence foods from Napakiak, a small Yup'ik Eskimo village on the Kuskokwim River, were assessed for total mercury content. The THg levels were low, ranging from 1 ng/g to 443.8 ng/g. A trend for the dried fish to have higher concentrations than the unprocessed fish was observed. Plants, as well as raindeer and moose meats, are low in THg and not associated with dietary exposure. Our data for Napakiak show that THg levels are low enough that mercury does not pose a significant health risk; however, more detailed consumption data in western Alaska is needed. PMID- 12650087 TI - "The demon in the freezer". PMID- 12650088 TI - [Diseases of civilization and a nosological principle of medicine in terms of general pathology]. AB - The last 100-150 years are marked by the spread of so-called "diseases of civilization" (primarily psychosomatic and mental diseases) which earlier have been encountered rarely. These diseases present quite novel pathogenic factors, adaptation to which in the course of evolution is lacking. Development of civilization constantly stimulates the emergence of numerous qualitatively new pathogenic factors, on the one hand, and changes the conditions for impact of conventional factors, on the other hand. This process is becoming more and more intensive and makes the sequence "cause-specific pathogenesis-clinical symptoms nosological unity-relevant etiotropic and pathogenetic treatment" less obvious. From this follow difficulties in application of routine methods of diagnosis, etiotropic and pathogenetic therapy based on the nosological principle as well as interest to methods of "alternative' (complementary) medicine which are nonspecific and, therefore, in some cases effective against some "diseases of civilization". PMID- 12650089 TI - [Genetics of arterial hypertension (current research paradigms)]. AB - Modern approaches to investigation of hypertension genetics are reviewed. These include genealogical, genetic-epidemiological, bioinformative methods, biological microchip technology. The results of genomic investigation of arterial hypertension are presented. Perspective lines in the research in the field of genetic cardiology are discussed. PMID- 12650090 TI - [The breath test is a necessary step in a physician's practice]. AB - The authors advocate introduction of the respiratory test into clinical practice, show its advantages over other methods of Hp diagnosis and the ability to diagnose non-invasively other gastrointestinal diseases especially in advanced infection primarily in viral hepatitides, HIV infections. The method is safe and comfortable. PMID- 12650091 TI - [Clinico-functional efficacy of medicinal and photon stabilization of cell membrane in patients with angina pectoris]. AB - Modification of erythrocytic membrane and the trend in clinicofunctional indices were studied in 90 patients with angina of effort (FC I-IV) in the course of treatment with a combination of membranoprotective drugs (group 1), magneto-laser radiation (group 2) and imitation of laser radiation (group 3). In patients of groups 1 and 2 the treatment resulted in stabilization of cell membrane accompanied with a hypotensive effect and increased exercise tolerance due to more effective cardiac performance. PMID- 12650092 TI - [Clinical course and intensive therapy of sepsis in geriatric patients]. AB - A comparative analysis was performed of a clinical course and intensive therapy of sepsis in 43 patients over 60 years of age. 86% of them were patients aged 60 to 74 years, the rest were older. The intensive therapy included: surgical treatment of the infection focus (on demand), antibiotics, immunomodulators (immunoglobulins or interleukine-2 preparations), nonsteroid antiinflammatory drugs (diclofenak), inhibitors of tumor necrosis factor (pentoxiphilline), vitamins of group B, infusion therapy, parenteral feeding (on demand), plasmapheresis or biospecific antiproteinase hemosorption. It is shown that geriatric patients develop sepsis due to chronic therapeutic pathology, their condition at admission to hospital is more severe. They have a changed posture of red blood and insignificant leukocytosis though the eukocytic index of intoxication in them is much higher. Biochemical indices in sepsis are approximately the same in all the age groups except total protein. Cellular immunity in the elderly is hyperactive while humoral immunity is depressed. Aged patients have hyperkinetic circulation and bad oxygen blood transport. A complex of therapeutic measures minimized sepsis lethality in geriatric patients. PMID- 12650093 TI - [Prognostic criteria of efficacy of inhalation glucocorticosteroids in non-severe bronchial asthma]. AB - The aim of the study was to evaluate efficacy of inhalation glucocorticosteroid (GCS) drugs in patients with persisting bronchial asthma (BA) previously steroid untreated and identify factors influencing the results of GCS therapy. 67 patients with non-severe BA given long-term theophilline with inadequate effect received inhalation GCS drug budesonid which produced a complete clinical effect (group 1), a delayed effect (group 2) or no effect (group 3). The above groups were compared by clinico-anamnestic data, functional respiration parameters, chemiluminescence of mononuclear cells of the peripheral blood measured before therapy with budesonide. The discriminant analysis estimated the minimal set of the initial parameters by which the groups differ. The regression model was used to calculate an individual prognostic index of probable treatment efficacy for each patient. The results of the study agree with the view on BA as a multistage disease in which chronic persisting inflammation provokes progressive remodeling of the respiratory tracts and formation of pathophysiological disorders resistant to glucocorticoids. It is necessary to use inhalation GCS drugs early in persistent BA. They are strictly indicated if BA patients need regular daily intake of short-term beta 2-agonists. PMID- 12650094 TI - [Clinical efficacy of combined anti-Helicobacter treatment of patients with duodenal ulcer]. AB - 84 patients with duodenal ulcer and positive by Helicobacter pylori (HP) were divided into two groups. Group 1 consisted of 42 patients who received omeprasol, metranidasol and clarithromycin. Group 2 of 42 patients received rovamycin instead of clarythromycin. The results of the study show that anti-HP regimen with rovamycin is superior to clarythromycin by main criteria (percent of healed ulcers, HP eradication, time to remission, recurrence rate, side effects rate). Thus, three-component therapy of patients with recurrent duodenal ulcer maintains intragastric acidity optimal for fast healing of ulcer defect. PMID- 12650095 TI - [The state of immune system and effects of current therapy and immunomodulators]. AB - To test the conception about a pathogenetic role of correlation between gastric comtamination with Helicobacter pylori (Hp) and immune system disorders in duodenal ulcer (DU), immune status and its response to current pharmacotherapy were studied in 84 patients in an acute recurrence phase. The patients were divided into 4 representative groups to receive monotherapy with omeprasol (group 1), combination denol + amoxicillin + tinidasol (group 2), quadrotherapy with drugs of group 1 and 2 (group 3), the above quadrotherapy plus immunomodulating drugs (imunofan + tactivin + oligovit) (group 4). In recurrent DU all the components of the immune system are affected especially cellular immunity. The response to the treatment was highest in group 1 and 4 (90% healing for 3 weeks), the worst in group 2 (68%). The complex of antibacterial drugs produces a moderate immunosuppressive action. The addition of immunomodulating drugs to the quadrotherapy promotes correction of secondary immunodeficiency and secondary Hp resistance to antibacterial drugs. This leads to a rise in eradication effect from 33-55 to 84% and a decrease in the number of early recurrences (during the first 6-12 months) from 42.1-33.6 to 12.5%. PMID- 12650096 TI - [Electrogastroenterography in diagnosis of pyloroduodenal ulcers]. AB - The authors propose a noval technique of computer electrogastroenterography to study motor-evacuatory gastrointestinal disorders in patients with pyloroduodenal ulcer (PDU). Bioelectrical activity of the stomach, proxymal and distal portions of the small intestine was compared in 78 patients with uncomplicated and complicated PDU. This activity was found to vary with acid-production function of the stomach, presence or absence of complications (subcompensated and decompensated pyloristenosis). The proposed method can be applied both for diagnosis of the motor-evacuatory disorders and control of therapy in patients with uncomplicated PDU, adequacy of preooperative preparation in complication of ulcer with pyloristenosis. PMID- 12650097 TI - [Clinical response to etiotropic therapy of chronic viral hepatitis C]. AB - Matched patients with chronic hepatitis C (CHC) were treated with monotherapy with intron A, or its combination with ribavirin or pegintron combination with ribavirin. The comparison group consisted of patients with a natural CHC course who received no specific antiviral therapy. The highest treatment efficacy was achieved in use of pegintron with ribavirin. However, this combination was associated with more frequent CNS and skin side effects. Side effects in all the above groups regressed completely for 1-3 months after the treatment. PMID- 12650098 TI - [A prospective study of hypolipidemic therapy in young women after myocardial infarction]. AB - A 3-year prospective trial has been performed of a course of ischemic heart disease (IHD) in postmyocardial infarction women (PIW) on antianginal treatment alone vs women on hypolipidemic drugs. IHD ran a less severe course in young PIW than in menopausal IHD. Aggravation of IHD occurred more frequently in women with previous MI and in hypertension. Comparison of the two groups of women demonstrated that hypolipidemic drugs improved a course of IHD in the women. In menopausal women taking hypolipidemic drugs IHD improvement occurred 6.2 times more frequently while aggravation 2.3 times less frequently than in women on standard antianginal treatment. Young women benefited from hypolipidemic therapy 1.6 times more frequently. PMID- 12650100 TI - [Hypoparathyroidism and syndrome of hypocalcemia]. AB - Hypoparathyroidism (HP) is a syndrome caused by insufficient secretion or weak action of parathormone leading to hypocalciemia (HC) and intermittent tetanus. HP is the most frequent cause of HC but not a single one. HC may present in 30 etiopathogenic variants and, consequently, being a problem for many clinical disciplines. Practical aspects of etiopathogenesis, clinical and laboratory diagnosis and treatment of a hypocalciemic syndrome of different nature are considered. PMID- 12650099 TI - [Current principles of treatment of acid-dependent diseases]. AB - The paper reviews current treatments of acid-dependent diseases. The role of hydrochloric acid hypersecretion in pathogenesis of some gastroduodenal diseases is shown. Efficacy of antisecretory drugs from different groups is compared. PMID- 12650101 TI - [Differential diagnosis of congenital fibrosis of the liver and hepatoportal sclerosis]. AB - The article deals with differential-diagnostic criteria of inborn liver fibrosis and hepatoportal sclerosis. Congenital liver fibrosis is inherited and manifests at young age. It is characterized by marked hepatomegaly with stony dense liver, moderate splenomegaly with hypersplenism and portal hypertension in high activity of alkaline phosphatase and 5-HT, renal affection. Fibrosis locates around the biliary ducts. Hepatoportal sclerosis manifests more frequently in the adults with moderate hepatomegaly, marked splenomegaly with hypersplenism and predominance of anemia, high portal hypertension. Fibrosis is moderate and locates around portal vein branches in which thrombi are found. PMID- 12650102 TI - [Septic endocarditis in nonspecific aortic arteritis]. PMID- 12650103 TI - [Results of activities and tasks of the Medical Corps of the Military Forces of the Russian Federation]. PMID- 12650104 TI - [System of indicators and criteria for assessing the economic effectiveness of military treatment and prevention facilities]. AB - The development of approaches to evaluation of the activity of military treatment and-prophylactic institutions (TPI) is very important for the structural reorganizations in the RF MD departmental health service, for detection of "the zones of irrational use" of the resources and factors of servicemen' medical support organization. The authors propose the model of using the economic-and mathematical and statistical methods of information processing concerning the TPI activity that permits to evaluate the possibilities of current modelling of activity indices for TPI ranging. They also show the importance and variation of activity efficiency evaluation in order to construct the horizontal relations at the "market" of medical services. It will contribute to more effective segmentation of policlinic and hospital services rendered the servicemen. PMID- 12650105 TI - [Judicial aspects of the cooperation between the Medical Corps of the Russian Federation Military Corps and the International Committee on Military Medicine]. PMID- 12650106 TI - [Perspectives for developing preventive services in the army and navy]. PMID- 12650107 TI - [Inadequacy in treating pneumonia and ways of eliminating it]. PMID- 12650108 TI - [Minimally-invasive combined radiosurgical interventions in diagnosis and treatment of pulmonary artery thromboembolism]. AB - Basing on the analysis of results obtained during the investigation and treatment of 38 patients suspected of pulmonary artery thromboembolism (PAT) the algorithm of measures applied in our hospital was developed. Its purpose was to put the PAT diagnosis and treatment in order. In our investigations the leading treatment method was the regional infusion therapy conducted through the transformation of diagnostic catheterization of pulmonary artery trunk into the treatment intervention. With the purpose of embolus hydrodynamic destruction the regional thrombolysis was conducted under conditions of fast high-pressure perfusion. It was preceded by the mechanical recanalization of thrombosis zone by the distal part of probe in order to create more favorable perfusion conditions. To prevent the repeated PAT when the source of thromboembolism was revealed the cava-filter implantation in the infra-renal part of inferior cava was performed. PMID- 12650109 TI - [Treatment of false aneurysms of vascular anastomoses]. PMID- 12650110 TI - [Diagnosis, treatment and rehabilitation of patients with consequences from orbital fractures]. PMID- 12650111 TI - [Microbial contamination of gunshot wounds to the face and neck]. PMID- 12650112 TI - [Prospects for computer modeling technology of osteosynthesis in fractures of bones of the lower limbs]. AB - To improve the results of treatment of the patients with femur and shin bone fractures the digital processing of video-information and computer simulation of perosseous osteosynthesis were used. The roentgenogram analysis was conducted using the personal computer equipped with the tolls for image input and visualization and software for data processing. 235 patients with femur and shin bone fractures were treated. The method of perosseous osteosynthesis was used in 127 of them. The digital computer simulation of the operation was conducted in 75 patients. The method has allowed to improve the anatomic and functional results, to reduce the treatment duration, to decrease the number of complications and unsatisfactory outcomes, to optimize the method of perosseous osteosynthesis in the whole. PMID- 12650113 TI - [Disruption of function of gastrointestinal tract organs in victims of severe trauma]. PMID- 12650114 TI - [On the 180th anniversary of the first issue of "Military-Medical Journal"]. PMID- 12650115 TI - Older patients and the aging Columbia shuttle. Examining organizational accidents to prevent disastrous outcomes. PMID- 12650116 TI - Vitamin B12 deficiency. Recognizing subtle symptoms in older adults. AB - Vitamin B12 deficiency is a common but under-recognized, yet easily treatable disorder in older adults. Although several causes exist, food-cobalamin malabsorption is now believed to be the most common etiology. Complications of vitamin B12 deficiency are myriad, ranging from lethargy and weight loss to dementia. Causes of deficiency include failure to separate vitamin B12 from food protein, inadequate ingestion, absorption, utilization, and storage as well as drug-food interactions leading to malabsorption and metabolic inactivation. The roles of B12 deficiency, elevated homocysteine and elevated methylmalonic acid in various disease states are still evolving. Timely screening and replacement of vitamin B12 will help prevent many complications. PMID- 12650117 TI - Hypertrophic cardiomyopathy. Pathophysiology, diagnosis, and treatment. AB - Hypertrophic cardiomyopathy is one of the most common inherited cardiovascular diseases and is characterized by a heterogeneous appearance and natural history. Although previously thought to be a disease of the young, HCM is frequently diagnosed in patients over age 50. A careful history and physical examination and readily available non-invasive testing will diagnose most cases, but genetic testing can identify those not expressing the typical phenotype. Treatment of symptomatic patients is targeted toward improving LV diastolic function; for patients with the obstructive form HCM, treatment involves relieving outflow tract obstruction. Identification of those at risk for sudden death may require consideration of prophylactic defibrillator placement. PMID- 12650118 TI - Depression in the very old. Differences in presentation and approach to treatment: a case example. PMID- 12650119 TI - Antibiotic-resistant skin infections spreading among gay men, also in prisons. AB - An outbreak of staphylococcus skin infection, causing large boils that are difficult to treat because these bacteria are resistant to many antibiotics, has spread recently among gay men in some cities, and also in certain prisons. In the past this infection occurred mainly in hospitals. PMID- 12650120 TI - Building grassroots support for AIDS. AB - Citizen-action campaigns (like getting people to call Congress or their state representatives to support Medicaid, ADAP, or other program for access to medical care) could get much more response if they were made accessible to the millions of people who are already supportive but not already involved. We suggest several ways to make action alerts and other citizen organizing work better. PMID- 12650121 TI - Bush proposes near tripling of U.S. commitment on global HIV epidemic. AB - The presidential initiative to increase funding for global AIDS prevention and treatment is widely seen as a groundbreaking advance. But critical time will be lost if the U.S. focuses so heavily on building its own new system, instead of using multinational organizations like the Global Fund that are ready now. PMID- 12650122 TI - Contest promotes HIV & HCV awareness. PMID- 12650123 TI - Hepatitis C. A case study in patchwork care. PMID- 12650124 TI - Program targets HIV+ injection drug users. AB - A large intervention program designed specifically for HIV-positive injection drug users may provide HIV programs with a workable way to teach this population about safety measures. PMID- 12650125 TI - Who will pay for the adverse events resulting from smallpox vaccination? Liability and compensation issues. AB - This paper summarizes liability and compensation concerns surrounding the smallpox vaccination program announced by President Bush on December 13, 2002. The paper examines the nature of adverse health events that are likely to occur in connection with the smallpox vaccine, assesses the liability protections that have been established for organizations and individuals participating in the vaccination program, and discusses the compensation mechanisms being considered to address the damages incurred by volunteers who may suffer from adverse vaccine reactions. Specifically, the implications of the Federal Tort Claims Act, workers' compensation programs, and the creation of a new no-fault compensation fund are explored. PMID- 12650126 TI - [Position of the German Society of Psychotherapeutic Medicine on psychoanalytically founded psychotherapy in relation to inquiry by "Scientific Psychotherapy Consultants" of the Federal Physician Group 15 April 2002]. PMID- 12650128 TI - 8th Congress of the European Confederation of Medical Mycology. Budapest, August 25-27, 2002. PMID- 12650127 TI - [EDITORIAL]. PMID- 12650129 TI - Report from 24th CIA Symposium: Allergy Frontiers and Futures. Bermuda, November 1-6, 2002. PMID- 12650130 TI - 8th Alps-Adria-Danube Congress on Sexually Transmitted Diseases and Infections of the Skin. Kranjska Gora, November 15-17, 2002. PMID- 12650131 TI - [Code guideline for gastroenterology--a practical guide]. PMID- 12650132 TI - Benign hepatic tumours. AB - Benign hepatic tumours include a broad spectrum of regenerative and true neoplastic processes. Due to advances in imaging procedures like MRI, Cf-scan and ultrasound as well as progress in immunohistochemistry, the appropriate diagnosis is made ina high percentage of patients without laparotomy and resection. Most important in clinical practice is the differential diagnosis of focal nodular hyperplasia and hepatocellular adenoma because of the risk of rupture and bleeding in the latter. Cavernous haemangioma, the most common benign hepatic tumour, rarely needs treatment. The diagnosis of nodular regenerative hyperplasia is often missed and patients present with secondary complications and signs of portal hypertension that necessitate treatment. The main problem in angiomyolipoma is to distinguish it from malignant processes which do require treatment. Because of its clinical presentation, inflammatory pseudotumour is also sometimes confused with a malignant tumour. Therapeutic options are drug therapy or surgical resection. Benign haemangioendothelioma of the infant is rare but may cause life-threatening complications. Bile duct adenoma is an incidental finding that is not known to cause any symptoms whereas biliary cystadenoma is often symptomatic and may progress to cystadenocarcinoma and therefore needs resection. PMID- 12650134 TI - Bibliography of toxinology. PMID- 12650133 TI - Formoterol thrice weekly does not result in the development of tolerance to bronchoprotection. AB - BACKGROUND: Loss of bronchoprotection routinely follows regular treatment with beta2-agonists. There are no data on the effects on bronchoprotection for thrice weekly use of a beta2-agonist. METHODS: A double-blind, randomized, placebo controlled crossover trial was conducted to investigate the effects of thrice weekly administration of 12 microg of formoterol versus placebo on bronchoprotection against methacholine. As an expected positive control, formoterol 12 microg once daily was also evaluated. RESULTS: There was no significant difference versus placebo in the bronchoprotective effects of 12 microg of formoterol administered on day 8, following daily treatment for seven days or treatment every other day (analysis of variance P=0.34). However, a nonsignificant trend towards lower concentration of methacholine that caused a 20% fall in forced expiratory volume in 1 s developed only following the daily formoterol dosing. CONCLUSIONS: Thrice weekly dosing does not result in the development of tolerance to bronchoprotection against the direct acting stimulus methacholine. PMID- 12650136 TI - Aluminum floc formation. PMID- 12650137 TI - Defense cleanup: a quick and dirty review. PMID- 12650138 TI - Low birthweight linked to asthma. PMID- 12650139 TI - Report on an EFSD/MSD Travel Fellowship for Young Scientists. PMID- 12650140 TI - Report on an EFSD/MSD Travel Fellowship for Young Scientists. PMID- 12650141 TI - SNM highlights as history: 1979. PMID- 12650142 TI - AIDSVAX results: an answer, or just more questions? AB - Some time within the next few weeks, VaxGen is expected to announce the results of the Phase III trials of its first vaccine candidate based on a bivalent HIV envelope gp120, AIDSVAX B/B. Interpretation of these results will probably not be straightforward and will likely lead to many more unanswered questions. Following is a description of some of the key issues involved. PMID- 12650143 TI - Lack of adverse events associated with pre- and postnatal antiretroviral therapy exposure: a retrospective review of 43 children followed up at 4-7 years of age. PMID- 12650144 TI - Foreward: cloning debate. PMID- 12650145 TI - No human cloning: a social ethics perspective. AB - This Essay addresses the negative impact of human cloning on the family, and argues further that market incentives to develop and implement cloning techniques exploit and exacerbate socioeconomic inequities. It suggests that cloning should be prohibited internationally and examines possible routes to that aim. To begin with, it offers some reflections on the nature of moral argument, and on the role of religion in public debate. PMID- 12650146 TI - Cloning: ethics and public policy. PMID- 12650147 TI - Religious attitudes toward cloning: a tale of two creatures. PMID- 12650148 TI - The case against human cloning. PMID- 12650149 TI - Who is the parent in cloning? AB - In July 1996, a sheep named Dolly was born in Scotland. What makes Dolly's birth noteworthy is that she is the result of the first successful cloning attempt using the nucleus of an adult cell. The technique that led to Dolly's birth involved transferring the nucleus of a mammary cell from an adult sheep to the enucleated egg cell of an unrelated sheep with gestation occurring in a third sheep. The possibility of applying this technique to human reproduction raised concerns worldwide with several countries moving for an immediate bans on human cloning. In the United States, President Clinton requested that the National Bioethics Advisory Commission ("NBAC"), a multidisciplinary group composed of scientists, lawyers, educators, theologians, and ethicists study the implications of cloning and issue recommendations. The Commission consulted other scientists, ethicists, theologians, lawyers, and citizens with interests in this advancing technology and concluded that, "at this time it is morally unacceptable for anyone in the public or private sector, whether in a research or clinical setting, to attempt to create a child using somatic cell nuclear transfer cloning." This Article was included in a larger work prepared at the request of, and submitted to the Commission by, law professor Lori B. Andrews. Cloning through nuclear transfer will change the way we create and define families. This Article explores how existing law relating to parentage, surrogacy, egg donation, and artificial insemination may apply in the cloning context to clarify the parent-child relationship established through cloning. PMID- 12650150 TI - Latina AIDS risk climbs. PMID- 12650151 TI - Does an anterior open bite in the early mixed dentition cause you to consider any form of orthodontic treatment?. PMID- 12650152 TI - Anchorage for closing open bites. PMID- 12650153 TI - Giving credit where credit is due. PMID- 12650154 TI - More information needed on properties of magnets. PMID- 12650155 TI - Corticosteroid injections, physiotherapy, or a wait-and-see policy for lateral epicondylitis? PMID- 12650156 TI - Update from the International Society on Hypertension in Blacks. PMID- 12650157 TI - Prevention of disability in daily activities in older persons with knee osteoarthritis. PMID- 12650158 TI - Full facial protection reduces injuries in elite young hockey players. PMID- 12650159 TI - Aerobic exercise for lowering blood pressure: a metaanalysis. PMID- 12650160 TI - Nobel prize winner trading card (CIRCA 1952). Elie Metchnikoff. AB - Russian doctor and bacteriologist, born in Ivanowca in 1845. He began his studies in Kharkov, continuing them at the Universities of Giessen, Gothingen, and Munich, later being named Professor of Zoology in Odessa in 1870. In the Canary Islands, he completed some anthropological works, but dedicated himself especially to studies of marine fauna. In 1887, much taken by the work of Pasteur, he wrote to him asking for a position in his laboratories; in a short time he became one of the principal collaborators with the master, especially in works concerning bacteriology. These were an inspiration to him, and led him to his famous theory of phagocytosis, the defensive act whereby white blood cells protect an organism against pathogenic microbes. Metchnikoff supposed that old age was avoidable, and subscribed to the materialistic school of thought. He was awarded the Nobel Prize in 1908. (With the complements of the Jose Lopez Luis Cigarillo Factory, Tenerife). PMID- 12650161 TI - Assessing environmental changes in grasslands. PMID- 12650162 TI - Human cloning: brave new mistake. PMID- 12650163 TI - Cloning: a business without regulation. PMID- 12650164 TI - The demand for human cloning. PMID- 12650165 TI - [Off-label use of clopidogrel after coronary stent implantation in Germany: optional or mandatory?]. AB - Although the combined use of acetylsalicylic acid (ASA) and clopidogrel represents the standard in the 4-week treatment after coronary stent implantation, a discussion persists in Germany regarding clopidogrel use and the actual measures for cost containment in the healthcare system. Indeed, clopidogrel has not been approved in Germany for use after stent im-plantation. The prescription of clopidogrel in this context is thus considered off-label use. Consequently, patients with social healthcare insurance should pay for clopidogrel out of their own pockets. This regimen, however, bears the inherent risk that patients will not purchase the drug and thus not take it. This overview article describes the risk to patients when social healthcare insurance companies would not pay for clopidogrel after coronary stent implantation. Five international, prospective, randomized and control-led studies in 3,230 patients have shown that "double" antiaggregation, i.e. a combination of ASA and the thienopyridin derivative ticlopidin, compared with ASA alone clearly reduced death, myocardial infarction and the need for another PCI or CABG to a manifold extent. Due to its lower rate of side effects, clopidogrel has replaced ticlopidin since mid-1998. The clinical results of three prospective, randomized studies and seven single-center registries in approximately 14,000 patients have shown that the combination of ASA and clopidogrel results not only in a lower rate of side effects, but also in a significant rate reduction of cardiovascular events of ca. 50% (from 4.0% to 2.1%). Therefore, the combined use of ASA and clopidogrel should be prescribed to avoid (sub)acute stent thrombosis --even if this is not according to the rules of the German healthcare system. The threat of fines to physicians prescribing clopdiogrel at the expense of the social health care insurance is counterproductive and jeopardizes the life of the patients. PMID- 12650166 TI - Two models of human cloning. PMID- 12650167 TI - "Being human": cloning and the challenges for public policy. PMID- 12650168 TI - How reprogenetics will transform the American family. PMID- 12650169 TI - Reflections on human cloning. PMID- 12650170 TI - [Glaucoma: an ocular blood flow disorder?]. PMID- 12650171 TI - [Photodynamic therapy carefully destroys neovascularization of the retina]. PMID- 12650172 TI - [Tumour of the choroid with yellowish subretinal deposits]. PMID- 12650173 TI - [Visual impairment in bilateral arcus lipoides. Schnyder's corneal dystrophy]. PMID- 12650174 TI - Effect of gravity change on thrombopoiesis in mice. AB - It is reported that the stay in the space develops anemia, thrombocytopenia, and altered function and structure of red blood cell. The mechanism of these abnormalities was not clarified yet. Therefore, it is necessary to elucidate the mechanism of the effect of the gravity change on the thrombocytopoiesis, which plays the important role for the hemostasis, using animal models. The cloning of thrombopoietin (TPO), followed by the analysis of TPO and c-mpl (its cellular receptor) knockout mice confirmed its role as the primary regulator of thrombopoiesis. TPO has been shown to stimulate both megakaryocyte colony growth from marrow progenitor cells and the maturation of immature megakaryocyte to form functional platelet. This process includes the massive cytoskeletal rearrangement, such as proplatelet formation and fragmentation of proplatelet. In this study we have focused on the thrombopoiesis in mice those were exposed to gravity change by parabolic flight (PF). PMID- 12650175 TI - Effects of water temperature on cardiac autonomic nervous system modulation during supine floating. AB - It is well known that heart rate, oxygen uptake and body temperature during exercise in water are affected by water temperature, buoyancy, hydrostatic and so on. It has been reported that the central blood volume during immersion was affected by the increased external hydrostatic pressure and cold-induced peripheral vasoconstriction, and intrathoratic blood volume should be greater during cold than warm water immersion (Epstein, 1992). The purpose of this presentation study was to make clear heart rate, blood pressure, oxygen uptake and cardiac autonomic nervous system modulation during supine floating at water temperatures of 25, 35 and 41 degrees C. PMID- 12650176 TI - Influence of long-term head-down body position on innervation density in extremity blood vessels. AB - The aim of the present study was to quantitate and compare the density of nerve terminals (NTD), as well as of their synaptic vesicle population (SyVD) in saphenous and brachial vein and artery, obtained from rats maintained in the horizontal or head-down tilted (HDT) position for two weeks. The same technique was applied as that for the head-up tilt study. PMID- 12650177 TI - Effect of simulated microgravity on human lymphocytes. AB - During space flight the function of the immune system changes significantly. Several papers reported that postflight the number and the proportion of circulating leukocytes in astronauts are modified (Leach, 1992), the in vitro mitogen induced T cell activation is depressed (Cogoli et al., 1985; Konstantinova et al. 1993) and there are detectable differences in cytokine production of leukocytes as well (Talas et al. 1983; Batkai et al. 1988; Chapes et al. 1992). One of the possible modifying forces is the microgravity condition itself. Our aim was to analyse mechanisms responsible for changing leukocyte functions in low gravity environment. For terrestrial simulation of microgravity we used a Rotary Cell Culture System (RCCS) developed by NASA. We investigated the effect of simulated microgravity on separated human peripheral blood mononuclear cells (PBMCs). We detected the populations of different cells by antibodies conjugated to fluorofors using a Flow Cytometer. Since space flight reduces the number of peripheral blood lymphocytes (Stowe et al., 1999) we supposed that apoptotic (programmed cell death) processes might be involved. This hypothesis was supported by the result of our earlier experiment demonstrating that simulated microgravity increased the level of secreted Tumor Necrosis Factor alpha (TNFalpha, a known apoptotic signal molecule) significantly (Batkai et al. 1999). PMID- 12650178 TI - Effects of long-duration bed rest on structural compartments of m. soleus in man. AB - Magnetic resonance imaging (MRI), histomorphometry and electron microscopy of muscle demonstrate that long-term exposure to actual or simulated weightlessness (including head down bed rest) leads to decreased volume of antigravity muscles in mammals. In muscles interbundle space is occupied by the connective tissue. Rat studies show that hindlimb unloading induces muscle fiber atrophy along with increase in muscle non-fiber connective tissue compartment. Beside that, usually 20% of the muscle fiber volume is comprised by non-contractile (non-myofibrillar) compartment. The aim of the present study was to compare changes in muscle volume, and in muscle fiber size with alterations in myofibrillar apparatus, and in connective tissue compartment in human m. soleus under conditions of 120 day long head down bed rest (HDBR). PMID- 12650179 TI - Morphology changes in rat skeletal muscles after 19 day exposure to +2 G. AB - Effects of long duration hypergravity on m. soleus morphology characteristics are still unknown. Particularly, only one paper describes the size and myosin heavy chain profiles of rat hindlimb extensor muscle fibers after 2 weeks of +2 G exposure. Earlier it was shown that long term overloading induced changes in skeletal muscle fiber cross-sectional area (CSA), fiber composition, and imitochondrial content. The aim of our study was to evaluate quantitatively structural parameters of m. soleus contractile apparatus after 19 day exposure to +2 G overloading. PMID- 12650180 TI - Dystrophin in limb skeletal muscle fibers and creatinkinase leakage after acute +3 Gz acceleration in rhesus monkeys. AB - Intensive muscle tension induces significant blood accumulation of enzymes and structural proteins of the muscle origin. Altered macromolecular permeability of the sarcolemma is attributed to integrity of sarcolemmal cytoskeleton, mainly to dystrophin-sarcoglycan (DSG) complex. It is known that intensive tension of the antigravity extensor muscles is observed under conditions of gravitational overloading. We assumed that acute exposure to hypergravity would lead to serum accumulation of creatine phosphokinase (CK) associated with considerably altered integrity of the dystrophin layer in fibers of extensor muscles. PMID- 12650181 TI - Mixed longitudinal study (6 years) of bone mineral density, muscle mass and muscle strength in the leg of aged females--relations to gravitational load and aging. PMID- 12650182 TI - Bioglass (R) stimulation of embryonic long-bones in altered loading environments. AB - The 45S5 composition glass (Bioglass (R), USBiomaterials, USA) contains 45 wt% SiO2, 24.5 wt% CaO, 24.5 wt% Na2O and 6 wt% P2O5. It is a low SiO2 (45 wt%) and a high Na2O content glass that results in rapid ion exchange when exposed to physiological solutions. The change in local solution chemistry results in rapid nucleation (1-3 hours exposure) and subsequent crystallisation of hydroxyl carbonate-apatite (HCA) identical to naturally occurring bone mineral within a hydrated SiO2-rich layer developed on the glass surface. During HCA layer formation, proteins and other growth factors are absorbed into the layer. The synthetic bone mineral layer bonds to collagen, produced by the host tissue, which leads to a strong interfacial bond that is stronger than the host tissue (hard or soft) (Hench and West. 1996). In this study we show that the chemical effects responsible for enhanced bone cell proliferation and differentiation found in-vitro and in-vivo with Bioglass (R) may be sufficient to prevent the demineralisation of bone tissue that occurs as a result of ageing or exposure to a low loading environment i.e. implant/tissue stress-shielding, space-flight. PMID- 12650183 TI - New approaches for study of mechanisms of bone demineralization due to microgravity. PMID- 12650184 TI - Effects of lower body positive pressure on muscle sympathetic nerve activity response [correction of respopnse] to head-up tilt. AB - The benefits of lower body positive pressure (LBPP) are generally accepted for clinical treatment in medical emergencies caused by massive bleeding to maintain the systemic blood pressure. They are also used by NASA post spaceflight for preventing orthostatic hypotension in the astronauts. However, controversy still exists concerning the mechanisms underlying LBPP benefits. The purpose of this study was to test the hypothesis that the baroreflex-mediated enhancement in sympathetic activity would be attenuated by LBPP during an orthostatic challenge in humans. Specifically, we studied 1) the sympathetic activity responses by the microneurographic technique, using direct intraneural measurement of muscle sympathetic nerve activity (MSNA); and 2) the contributions of preload and afterload to the chances in MSNA response during orthostasis on application of LBPP. To accomplish these issues, MSNA was recorded microneurographically along with noninvasive measurement of the cardiovascular variables in all the subjects during exposure to a 70 degrees HUT with 30-mm Hg LBPP. PMID- 12650185 TI - Effect of long-term bedrest on lower leg muscle activation patterns during quiet standing. AB - It has been well known that balance instabilities after long-term exposure to microgravity (e.g., Anderson et al. 1986) or bedrest (BR) can be related to alterations and/or adaptations to postural control strategies. Little is known, however, how the reduced muscular activity affects the activation pattern of the lower limb muscles during quiet standing (QS). The purpose of this study was to investigate whether or not any changes in the lower limb muscle activation patterns during QS would occur after BR. PMID- 12650186 TI - Morpho-functional adaptations in the bone tissue under the space flight conditions. AB - Microgravity in space flight--situation of a maximum deficit of supporting loading on the skeleton and good model for finding-out of osteopenia and osteoporosis development laws, which are wide-spreading now and are "civilization diseases". Most typical for bones in conditions of a microgravitation by changes are: a decrease of intensity growth and osteoplastic processes, osteopenia and osteoporosis, decreasing of a mechanical strength and the risk of breaches arising (Oganov V.S., Schneider V. (1996)). Cytological mechanisms of gravity dependent reactions in a bone tissue remain in many respects not-clear. By the purpose of our work was the analysis of some ultrastructural changes in bone tissue cells of the monkeys (Macaca mulatta), staying during two weeks onboard the biosatellite BION -11. PMID- 12650187 TI - Vestibular and somatosensory interaction during recovery of balance instability after spaceflight. AB - The aim of our contribution was to characterize vestibular and somatosensory influence on human balance recovery during readaptation to the earth conditions after spaceflight. We tested how post-spaceflight postural reactions to galvanic stimulus (related to vestibular input) and to vibration of the lower leg muscles (somatosensory input) were changed. PMID- 12650188 TI - Simulated microgravity induces alteration in the central nervous system. AB - To investigate whether the signs of neurophysiological impairment observed in flight may be traced back to cytomorphology, we undertook a ground-based study focusing upon the architecture of cultured glial cells under simulated microgravity obtained by three-dimensional clinorotation. PMID- 12650189 TI - Reflex response changes during hyper and microgravity. AB - This paper reports the quantitative evaluation of the H-reflex exhibited by parabolic flight with exposure to micro and high-gravity. With respect to previous findings in parabolic flights and short-term space missions, the analysis focused on reflex activity in weightlessness. The aim of this study was to investigate the effect of gravity on H-reflex and motor evoked potentials (MEP) in soleus muscle (SOL) during parabolic flight. PMID- 12650190 TI - Simulation and interpretation of experiments with otoliths. AB - The function of otolith as a gravisensor comprises a sequence of different physical and chemical processes, which experimental investigation is difficult or unfeasible. To understand the relationship between the input and output in otolith organ, certain assumptions concerning the mechanisms of transformation of information on the inner, intermediate stages have to be made. Their validity has to be tested by comparison of the properties of output characteristics, which follow from these assumptions, with experimental data. The goals of the present paper are: 1) to analyze some of the assumptions used in and related to the intermediate stages of transformation of mechanical stimulus in neural response; 2) to demonstrate by comparison with the results of modeling of otolith structure that some peculiarities of the experiment may be caused by spatial inhomogeneity of otolithic membrane (OM). PMID- 12650192 TI - Model of statoconia accumulation in gravireceptors of mollusks. AB - The kinetics of formation and accumulation of statoconia are different for Aplysia californica and Biomphalaria glabrata. In Aplysia californica, the fast growth of statoconia number occurs after the critical size (approximately 45 micrometers) of statocyst is reached; then the increase of statoconia number is proceeding with the nonmonotonic rate during the life of an animal. In Biomphalaria the growth of statoconia number occurs only in the initial phase. Then long-term evolution of statoconia in the absence of their generation is the result of their growth in the cyst lumen. In the case of Aplysia californica it is not clear whether a temporal change of the statoconia size distribution (SSD) is caused by statoconia growth in the cyst lumen similar to that in Biomphalaria (Model 1) or statoconia growth takes place in supporting cells until their release into the cyst lumen occurs. (Model 2). This problem is of practical importance because the majority of experiments related to the development of molluscan gravireceptors in altered gravity dealt with an initial phase of statoconia evolution in Aplysia californica and Biomphalaria glabrata. The purpose of the present work is the application of mathematical modeling to the analysis of mechanisms of statoconia formation by supporting cells. PMID- 12650191 TI - The effects of hypergravity on vestibular epithelia and behaviour of the rat. PMID- 12650194 TI - The vestibulo-ocular reflex of hypergravity rats. AB - The vertebrate vestibular system detects linear (otolith organs) and angular (semicircular canals) acceleration. The function of the otolith system is twofold, 1: perception of linear acceleration of the head, and 2: assessment of the spatial orientation of the head relative to the vector of gravity. Because of the latter function, a change of gravity will affect the vestibular input which, in turn, may have a wide range of serious physiological effects, for instance on ocular reflexes. The function of the vestibulo-ocular reflex (VOR) is to stabilize the visual image on the retina. Measurement of this VOR provides a method to investigate the (processing within the) vestibular system. Discrimination between gravity and linear acceleration, caused by movement of the head, is not possible. Therefore, information from the otolith system must be constantly compared with additional information from other sensory systems in order to solve the inherent ambiguity between tilt and translation. In this processing, cues from the semicircular canals also play a role. During parabolic flight, experiments can be performed at altered gravity levels for brief periods of time. On earth, the only effective possibility to manipulate gravity for longer periods of time is a centrifuge. Together with experiments in weightlessness during orbital flight, these methods form useful tools to investigate the influence of gravity on physiology. In our laboratory, rats have been kept inside a centrifuge at 2.5 g during their entire life-span (i.e. including gestation). PMID- 12650193 TI - Hypergravity and opioid-mediated pain suppression in rats. AB - It is known that pain suppression in animals is induced by certain environmental stimulus. However, little is known about the effects of gravitational alteration on the nociceptive responses in rats. A recent study indicated that Fos protein expression was strongly induced in the vestibular-related brainstem regions of rats that were exposed to 2 G hypergravity (Gustave Dit Duflo et al., 2000). A number of studies indicate that Fos expression is induced in the brain by various kinds of stress. We showed that either long-term exposure or short-term exposure to 2 G hypergravity elevated the nociceptive threshold in the rat skin surfaces, in concomitant with Fos induction in the hypothalamus including the arcuate nucleus and paraventricular nucleus (Kumei et al., 2000). We have examined the possible involvement of beta-endorphin, an endogenous opioid, in the hypergravity induced analgesic effects on rats and its counteraction by naloxone, an opioid receptor antagonist. PMID- 12650195 TI - Independent dendritic areas give rise to compound, nonlinear and phase-sensitive integration of input information in the cerebellar Purkinje-cells. PMID- 12650196 TI - Endocrine responses to space flights. AB - Simultaneously with human space flights several series of observations were performed by using experimental animals--mainly rats--exposed to space flights on board of special satellites BION-COSMOS or in Shuttle Transportation Systems (STS). The aims of these experiments were to study in more details: the mechanisms of the changes in bones and skeletal muscle, the alterations of the function of immune system, the radiation effects on organism, the mechanism of the changes of endocrine functions, the evaluation of the role of hormones in alteration of metabolic processes in organism. The advantages of these animal experiments were the possibilities to analyze not only the plasma samples, but it was possible to obtain samples of organs or tissues: for morphological and biochemical analysis for studies of the changes in enzyme activities and in gene expressions, for measurement of metabolic processes and for investigation of the hormone production in endocrine glands and estimation of the response of tissues to hormones. It was also possible to compare the endocrine response to spaceflight and to other stress stimuli. These animal studies are interesting for verification of some hypothesis in the mechanism of adaptation of human organism to the changes of gravity. The disadvantage was, however, that the animals in almost all experiments could be examined only after space flight. The actual inflight changes were investigated only in two SLS flights. In this short review it is not possible to evaluate all hormonal data available on the response of endocrine system to the conditions of space flights. Therefore we will concentrate on the response of pituitary adrenocortical system, pituitary thyroid and pituitary gonadal functions. PMID- 12650198 TI - Lipoxygenase activity during parabolic flights. AB - Experiments in Space clearly show that various cellular processes, such as growth rates, signaling pathways and gene expression, are modified when cells are placed under conditions of weightlessness. As yet, there is no coherent explanation for these observations, though recent experiments, showing that microtubule self organization is gravity-dependent suggest that investigations at the molecular level might fill the gap between observation and understanding of Space effects. Lipoxygenases are a family of dioxygenases which have been implicated in the pathogenesis of several inflammatory conditions, in atherosclerosis, in brain aging and in HIV infection. In plants, lipoxy-genases favour germination, participate in the synthesis of traumatin and jasmonic acid and in the response to abiotic stress. Here, we took advantage of a fibre optics spectrometer developed on purpose, the EMEC (Effect of Microgravity on Enzymatic Catalysis) module, to measure the dioxygenation reaction by pure soybean lipoxygenase-1 (LOX 1) during the 28th parabolic flight campaign of the European Space Agency (ESA). The aim was to ascertain whether microgravity can affect enzyme catalysis. PMID- 12650197 TI - Effects of 17 days of head-down bed rest on hydro-electrolytic regulation in men. AB - Prolonged periods of head-down bed rest (HDBR) are commonly used to mimic the effects of microgravity. HDBR has been shown to produce, as in space, a cephalad redistribution of circulating blood volume with an increase in central blood volume which induces the early adaptations in blood volume regulating hormones. Changes in atrial natriuretic peptide (ANP), arginine vasopressin (AVP), renin activity and aldosterone have been observed. Many reports describe these endocrine adaptations but few investigations of rhythms are in the literature. We proposed to evaluate the circadian rhythms of the hormones and electrolytes involved in the hydro-electrolytic regulation during a HDBR study which was designed to simulate a 17-day spaceflight (Life and Microgravity Spacelab experiment, LMS, NASA). PMID- 12650199 TI - Modification of urinary secretion of 8-hydroxy-2'-deoxyguanosine and serum ACTH concentration following repetitive parabolic flights. AB - It is important to clarify the molecular mechanisms of physiological responses of the human body to changes in gravity. Previous reports demonstrated that gravity changing stress increases the human urinary concentration of 8-hydroxy-2' deoxyguanosine (8-OHdG). However, it has yet to be clarified whether repetitive parabolic flight modulates the urinary concentration of 8-OHdG after exposure to gravity-changing stress. In the present study, the effects of the number of previous experiences with parabolic flight on urinary excretion of 8-OHdG and concentration of serum ACTH were examined in 12 healthy volunteers. PMID- 12650200 TI - Effect of simulated microgravity conditions on poly(ADP-ribose) polymerase activity in primary cultures of adult rat hepatocytes. AB - The possible involvement of poly(ADP-ribose) polymerase [PARP; E.C. 2.4.2.30] in the adaptive response to low-g conditions was studied in cultured adult rat hepatocytes exposed to simulated microgravity produced by the random positioning machine (RPM-3D-clinostat). Four different poly(ADP-ribose) polymerases (PARPs) have been identified recently. The best-studied member of this family is PARP-1, a highly conserved, multimodular 113 kDa protein. In multicellular organisms PARPs catalyze poly(ADP-ribose) synthesis from NAD+ to a number of structural and catalytic proteins. Moreover, PARP-1 can control its protein and DNA interactions by catalyzing its automodification with poly(ADP-ribose) molecules that can include up to 200 ADP-ribose residues and several branching points; by these polymers, PARP-1 may nocovalently interact with other proteins and alter their functions. PARP-1 binds to DNA and is activated by free ends interacting with several other DNA damage checkpoint proteins. Thus, PARPs may target specific signal network proteins via poly(ADP-ribose) and regulate their domain functions. Poly(ADP-ribosyl)ation plays a central role in genome stability and is involved in DNA replication and repair, gene expression, cell differentiation and transformation. We have shown that a loss of PARP-1 activity is a critical event in the early molecular steps of the hepatocarcinogenesis process. Moreover, a prompt increase in this enzymatic activity is linked not only to the presence of DNA free ends but is linked also to the start of DNA synthesis. More recently, we have reported that PARP-1 is involved in hormone-mediated gene expression in vitro and in vivo during rat liver regeneration. PMID- 12650201 TI - Effect of microgravity on plasma catecholamine responses to stressors during space flight. AB - The effect of microgravity on the sympathicoadrenal system (SAS) activity in humans and animals has not yet been clarified. Our previous studies suggested that the SAS activity, evaluated by circulating and/or urinary catecholamine (CA) levels in astronauts during space flights, was found to be rather unchanged. However, CA levels were measured in astronauts only at rest conditions. The aim of the present study was to investigate effect of microgravity during space flight and post-flight readaptation on responsiveness of the SAS to somatic and psychic stressors evaluated by levels of catecholamines and their metabolite in the blood of the Slovak cosmonaut during his stay on board the space station Mir. PMID- 12650202 TI - Plasma hormone levels in human subject during stress loads in microgravity and at readaptation to Earth's gravity. AB - In great part of the investigations of endocrine system functions in astronauts during space flights the plasma levels of hormones and metabolites were determined only in resting conditions, usually from one blood sample collection. Such levels reflected the psychical and physical state and new hormonal homeostasis of organism at the time of blood collection, however, the functional capacity of neuroendocrine system to respond to various stress stimuli during space flight remained unknown. The aim of present investigations was to study dynamic changes of hormone levels during the stress and metabolic loads (insulin induced hypoglycemia, physical exercise and oral glucose tolerance test) at the exposure of human subject to microgravity on the space station MIR. The responses of sympatico-adrenomedullary system to these stress and workloads were presented by Kvetnansky et al. PMID- 12650204 TI - Microgravity and aging of animals. AB - A study of changed gravity effects upon viability, life span and aging is of interest, on one hand, from a practical viewpoint in relation to the growing duration of space missions and on other hand, from a theoretical viewpoint, because gravity is one of the key factors in the evolutionary process on the Earth. In 1978 special conference titled "Space Gerontology" was held. Well known experts in space biology and physiology of aging participated in it. However, all the materiales presented at the conference were based on analogies and on what could be during exposure to microgravity rather than on real data. I shall try to discuss this problem, basing on the results of rats experiments on board "Cosmos" biosatellites and ground based model experiments. Male wistar rats examined after 1-3 weeks exposure to microgravity on board biosatellites demonstrated some changes similar to the signs [correction of sings] of aging, such as decreased motor activity, thymus involution, muscle atrophy, osteoporosis etc. But all these changes were reversible and in rats examined 3 weeks after return to the Earth we did not find any deviations from the controls. PMID- 12650203 TI - Altered gravity effects on mothers and offspring: the importance of maternal behavior. AB - In this paper, I review and discuss recent studies of pregnant, parturient and lactating rat mothers and neonates exposed to hypo- and hypergravity. These studies are revealing new insights into how deviations form Earth-normal gravity may affect fundamental reproductive and ontogenetic processes in mammals. By way of background, I will first briefly summarize the spaceflights that have carried mammalian mothers and their offspring into space. PMID- 12650205 TI - Hypergravity studies in the Netherlands. AB - It looks like that with the utilization phase of the International Space Station (ISS) scientists will have the possibility to perform long duration and more sophisticated microgravity experiments than could be performed previously. In preparation for these spaceflight studies, ground based experiment tools for simulated (or real) microgravity and hypergravity are important. To provide the infrastructure and user support necessary to perform these ground based studies we have setup the Dutch Experiment Support Center, DESC. This paper will focus on the three Dutch centrifuge facilities. It is shown that these hypergravity facilities can be used to show sounding rocket launch effects, identify alterations in body mass, bone parameters and matrix composition in rodents as well as to derive a test protocol for the Space Adaptation Syndrome in humans. DESC coordinates the use of these centrifuge facilities. PMID- 12650206 TI - 3D realtime echography and echography assisted by a robotic arm for investigating astronauts in the ISS from the ground. AB - As human will stay for long duration in isolated sites like ISS there will be a need to perform quick and reliable diagnosis to evaluate the gravity of the pathology in presence of clinical symptoms. Many pathological situations (abnormal heart rate, pericardic collection, mitral prolaps, cholecystis, renal lithiasis, normal and ectopic pregnancies, ovarian cyst, acute appendicitis, phlebitis ... ) may occur even if all the astronauts are absolutely normal and healthy preflight. Ultrasound echography and Doppler are non invasive methods easy to use in space and very well adapted and used in routine for such diagnosis at the hospital. The objective of the present project was to design a method that guarantee a reliable echographic diagnostic in an isolated site (space station or earth site) by a Medical Doctor located at the expert site that should be the Nasa control center for ISS. It is supposed that there is only a non sonographer person in the isolated site and that the transmission system (audio, video, numeric..) is the only link between the 2 sites. Two options are proposed: (a) A 3D realtime acquisition echograph that can record quickly all the echos of a volume containing the organ suspected to have a lesion, all these echo information being sent to the ground and processed by the ground experts, (b) A robotic arm that hangs the echo probe in the isolated site tele-operated (through sattelite network) from the ground by an expert in clinical ultrasound. (As the expert moves the joystick of his ground computer the robotic arm reproduces the same movements on the probe). PMID- 12650207 TI - Modern view on the short-arm centrifuge as a potential generator of artificial gravity in piloted missions. AB - The problem of artificial gravity (AG) in long-term missions is one of the hottest, as the existing countermeasures do not fully cope with the negative consequences of weightlessness. From two variants of AG creation--rotation of space systems around of their mass center or short radius centrifuge (SAC)--the preference is given to SAC, as technically easier variant for realization. However, the rotation of a person on SAC can cause not only positive, but also negative effects. The purpose of the present study was to perform the analysis of data of researches on a problem of AG generated by a of SAC, executed in Russia during last 20 years, and to state modern views on application of SAC as a mean of AG creation in long piloted missions. PMID- 12650209 TI - Bibliography of toxinology. PMID- 12650208 TI - LBNP as useful tool for pilot candidates selection to the Polish Air Force: a preliminary study. AB - BACKGROUND: Candidates to the Polish Air Force are to fulfill strict medical conditions. Every candidate, wanting to become a fighter pilot or not, has to pass a rigorous centrifuge examination. This is a costly and especially stressful process for the candidates, as first-timers. This article explores possibility for different approach to candidates fitness to fly assessment. METHODS: 14 candidates (21-22 yrs old) were subjected to a regular GOR centrifuge profile (0.1 G/s; up to +7.0 Gz; unprotected) and to lower body negative pressure (LBNP) examination (2 min. rest period followed by 3 min. at -40 mm Hg; supine position). Both, centrifuge and LBNP examinations were carried out the same day, with about 1 hour gap between them. ECG and impedance cardiography (ICG) were being recorded during both tests. The following parameters were calculated: HR and QZ (time between ventricular depolarization [beginning of the Q wave in ECG] and local maximum of the first derivative [dZ/dt] in ICG). QZ was considered to be an effective measure of dynamic heart function. RESULTS: +Gz tolerance for the group was +5.94 +/- 0.98 Gz. Correlation between +Gz tolerance and QZ values during LBNP test (-40 mm Hg) for all subjects was equal to r=0.6. Correlation between sets of parameters (canonical analysis) for LBNP and centrifuge was equal to r=0.78. Both results were statistically insignificant (p<0.09). Baseline HR values were 121 +/- 19 bpm and 75 +/- 12 bpm, and maximal HR values were 172 +/- 21 bpm and 81 +/- 13 bpm, respectively for centrifuge and LBNP. For both, baseline and maximal HR during LBNP and G exposure canonical correlation was equal to r=0.72. The results were statistically not significant, however. CONCLUSIONS: Centrifuge examination is much more stressful than LBNP test. This could have implication on the objective results of centrifuge test. The results may suggest that LBNP test could be a used in pilot candidates selection, especially those presenting high baseline HR values. PMID- 12650210 TI - Cochrane reviews-pulse oximetry for monitoring. PMID- 12650211 TI - [Ventricular assist system]. PMID- 12650214 TI - [Molecular oncogenetics, oncogenomics]. PMID- 12650215 TI - Suicide and aircraft. PMID- 12650216 TI - Incidence of completed and attempted suicide in Trabzon, Turkey. AB - OBJECTIVE: Studies of completed and attempted suicide in Turkey are based on data of State Institute of Statistics (SIS) and emergency clinics of the large hospitals. This study seeks (1) to find, independent of the SIS and hospital data, the annual incidences of completed and attempted suicide in Trabzon, Turkey; (2) to examine the associated factors between the incidence of completed and attempted suicide. METHOD: The data are derived by using a method specially designed for this study. Data sources include emergency clinics in all hospitals, village clinics, the Forensic Medical Center of Trabzon, the Governorship of Trabzon, "mukhtars" (local village representatives) of neighborhoods, the Office of the Public Prosecutor of Trabzon, the Police Headquarters and Gendarmerie, and the local press organs. RESULTS: The incidences of completed and attempted suicide per 100,000 inhabitants turned out to be 2.60 and 31.5, respectively, whereas the SIS reported the incidence of completed suicide to be 1.11 per 100,000 inhabitants in Trabzon in 1995. CONCLUSION: Our results demonstrate that SIS data are inadequate for suicide research in Turkey. Our findings show that the risk of completed and attempted suicide is high in young, unmarried, and unemployed persons, and that these groups must be carefully evaluated for suicide risk. The study highlights the need for culture-specific research on suicidal behavior in Turkey. PMID- 12650217 TI - Suicide and suicide attempts in Ankara in 1998: results of the WHO/EURO Multicentre Study of Suicidal Behaviour. AB - BACKGROUND: The rates and associated basic demographic features of attempted and completed suicides in a catchment area in Turkey were investigated as part of the WHO/Euro Multicentre Study of Suicidal Behaviour. METHOD: All hospitals in the catchment area were screened in order to identify attempted suicides. Statistics for completed suicides were obtained from the State Institute of Statistics (SIS). RESULTS: The rates of attempted and completed suicides per 100,000 inhabitants over 15 years of age were 31.9 for males and 85.6 for females, and 9.9 for males and 5.6 for females, respectively. The majority of attempted suicides were in the 15-24-year-old age group, as is the case in all other European countries. The majority of completed suicides were also in the 15-24 year-old age group, although in other European centers most completed suicides occur in the 40+ age group. The most frequent methods were overdose for attempted suicide and hanging for completed suicide. The rates of both attempted and completed suicides were lower than those of other participating centers in Europe. CONCLUSION: Male sex is a risk factor for completed suicide and female sex is a risk factor for attempted suicide, while an age of 15-24 years may be a risk factor for both groups. PMID- 12650218 TI - The protective role of parental involvement in adolescent suicide. AB - This study of 2,722 adolescents aged 14-18 years explored whether parental involvement can protect against adolescent suicide attempts. Compared to their counterparts suicide attempters were more likely to have been in trouble with the police, to report lower levels of parental interest and academic motivation, and to report suicidal ideation and using alcohol or an illegal drug when they feel stressed. They were also less likely to reside with both parents. The association between parental involvement and suicidal behaviour was not stronger for sons than for daughters or for adolescents who had experienced family disruption than for those who grew up in two-parent families. PMID- 12650219 TI - Struggling against suicide: the need for an integrative approach. AB - Transcultural comparisons would be beneficial for both our understanding of suicide phenomena and the setting up of preventative strategies. Changes in attitude throughout history, gender and age differences, socio-economic factors, influence of race and ethnicity, and the impact of religion are among the aspects mentioned in this article. All of them can actually have a considerable effect on suicide rates, and all of them should be taken into account when interpreting suicide trends and designing antisuicide strategies. In fact, both risk and protective factors may vary remarkably in different cultures. The need for an integrative approach is evident today more than ever, and this fact calls for more concerted and coordinated, multidisciplinary approaches to suicide prevention. PMID- 12650220 TI - Suicide in the Ukraine. AB - The suicide rates of the 24 provinces (oblasts) of the Ukraine were found to be strongly associated with indices of social disintegration (such as divorce and illegitimacy rates), with the Western provinces incorporated in the USSR later than other Ukrainian territories having lower suicide rates and lower levels of social disintegration. PMID- 12650221 TI - Sophocles--the first great suicidologist. PMID- 12650222 TI - Grandiose fury. Letter across the pacific. PMID- 12650224 TI - I.A.S.P. Conference Report, Chennai, September 2001. PMID- 12650223 TI - Assisted suicide, euthanasia, and the right to end-of life care. PMID- 12650225 TI - Postconcussive symptom report: the relative influence of head injury and depression. AB - The present study explored whether any subset of self-reported postconcussion (PCS) symptoms or specific PCS symptom is sensitive and/or specific to head injury in non-self-selected samples of individuals aged 18-21 with head injury and depression (n = 32), head injury without depression (n = 31), depression without head injury (n = 25), and controls (n = 50). All participants completed a self-report PCS symptom scale based on their current symptoms. Results showed that depression, not head-injury status, largely accounted for elevation in PCS symptom reports, including cognitive symptoms. Thus, report of cognitive PCS symptoms is not specific to head injury, raising concerns about using such items to screen for head injury in the general population. PMID- 12650226 TI - Controlled processes and automaticity in memory functioning in fibromyalgia patients: relation with emotional distress and hypervigilance. AB - Evidence exists that chronic pain partially consumes the limited attentional resources, with the consequence that controlled processes sustaining cognitive tasks are affected and that automatic processes are preserved. Fibromyalgia syndrome is consistently rated as more severe than other chronic painful conditions. It is assumed here that fibromyalgia is more attention-demanding, leading to a more pronounced decrease of the controlled processes in comparison with other chronic painful conditions. In this perspective, Study 1 compares fibromyalgia patients, patients with localized pain and healthy subjects in a procedure separately estimating the within-task contributions of controlled and automatic processes in a cued recall task. As predicted, controlled processes are more strongly affected in fibromyalgia patients related to the group with localized pain. Unexpectedly, contribution of automatic processes is increased in fibromyalgia. Study 2 replicates these results and reveals that memory functioning in fibromyalgia patients is related to their painful condition as a whole rather than to any particular patient's characteristics. PMID- 12650228 TI - Inhibition of expected movements in Tourette's Syndrome. AB - Tourette's Syndrome (TS) is a basal ganglia (BG) disorder characterized by involuntary motor and vocal tics, impairment of voluntary motor function, and attentional dysfunctions. This experiment aimed to investigate motor reprogramming and inhibitory control in adult TS patients. Ten TS patients and 10 age- and sex-matched controls performed sequences of movements in response to the illumination of LED lights, and were required to interrupt a predictable, reciprocating sequence of leftward and rightward movements in order to move to an unpredictable target that occurred either in the direction opposite to that expected, or in the opposite direction and at twice the expected distance. Results showed that TS patients were no slower than controls in reprogramming the direction and distance of movements, thus showing no overall deficits in inhibitory control. Of interest, however, was a directional asymmetry (right bias) in movement preparation time in TS patients. Patients with TS may have a superiority in allocating attentional resources toward the right side, contralateral to the implicated reduced BG volume in this disorder. These findings extend earlier work reporting anomalous BG lateralization in TS. PMID- 12650227 TI - Sensory-perceptual dysfunction in patients with schizophrenia and comorbid alcoholism. AB - We studied the effects of aging and comorbid alcoholism on sensory-perceptual dysfunction in patients with schizophrenia. Using sensory-perceptual tests from the Halstead-Reitan Neuropsychological Battery included in the Russell, Neuringer, and Goldstein (1970) Perceptual Disorders Index, samples were compared of 54 patients with comorbid schizophrenia and alcoholism, 234 patients with schizophrenia but no history of alcoholism, 132 patients with chronic alcoholism and 171 patient controls. Data were analyzed by one-way analyses of variance followed by Scheffe multiple comparison tests. Numerous significant differences were found among groups, with the comorbid schizophrenia group doing more poorly than the other groups on measures of touch perception and suppression, finger agnosia, and fingertip number writing. Multiple regression analyses using the sensory-perceptual tests as predictor variables and age as the dependent variable indicated that the strength of association between age and the multivariate set of sensory-perceptual test scores was greater in the comorbid schizophrenia group than the noncomorbid schizophrenia, alcoholism and patient control groups. The preponderance of age-related deficits was left-sided, suggesting accelerated decline in right hemispheric function. A comparison of groups across ages on a single Perceptual Disorders Index score showed a substantially larger age effect in the comorbid schizophrenia group relative to the other groups. PMID- 12650229 TI - Subjective changes in emotion, cognition and behaviour after stroke: factors affecting the perception of patients and partners. AB - The presence and severity of changes in emotion and cognition experienced by left and right-sided stroke patients and observed by their partners were compared at 3 months poststroke. The results showed that, regardless of the side of stroke, several changes were reported by half of the stroke patients and their partners. It appeared that while left hemisphere stroke patients agreed with their partners on the number and severity of most changes, partners of right hemisphere patients reported more frequent and more severe changes than the patients themselves. The level of observability of the altered behaviour, distress of the partner, distress of left-sided stroke patients and hemispatial neglect of right-sided stroke patients emerged as factors related to disagreement between stroke patient and partner. PMID- 12650230 TI - Reduced task-set inertia in Parkinson's disease. AB - This study investigated the relationship between cognitive inhibition and set switching costs in 13 patients with Parkinson's disease (PD) and 16 control subjects using a set-switching task with fully predictable switches. Incongruent colour-words and numerals were presented in either baseline (only colour-words or only numerals) or alternating lists (colour-words and numerals in alternation). Words required either a word-reading or colour-naming response whereas numerals required either a value-naming (value of the digits) or group-size naming (number of digits) response. Stroop interference was found to be increased and practice effects on nondominant tasks reduced in PD patients, suggesting that the ability to increase control over interference from irrelevant prepotent responses may be implicated in PD. Switch costs of PD patients were reduced on the first few trials of alternating dominant task lists and increased towards the end of alternating nondominant task lists. Both effects were explained as resulting from a reduced ability to maintain task-sets where selection of correct responses relies on inhibition of irrelevant stimulus-response mappings. PMID- 12650231 TI - Exploring the effect of action familiarity on SPTs recall performance in Alzheimer's disease. AB - This study examined the performance of normal controls (NC) and Alzheimer's disease (AD) patients on free recall, semantic cued recall and object cued recall of both subject-performed tasks (SPTs) and verbal descriptions of actions, by controlling familiarity of actions associated to objects. The results showed that both groups performed better after SPT encoding than after verbal encoding, in all three types of recall. In addition, this SPT advantage was greater for AD patients than for NC in the object cued recall test, emphasizing AD patients' sensibility to the congruence of cues between encoding and retrieval conditions. Following verbal encoding, NC showed a better recall for less familiar actions than for highly familiar actions, whereas AD patients exhibited the opposite pattern. These results reflect that AD patients did not benefit from a distinctiveness effect at encoding for improving subsequent retrieval of verbal information, probably due to a reduced level of elaboration during encoding. However, there was no effect of action familiarity on recall performance by both groups following SPT encoding. These results suggest that memory for verbal actions and SPTs is governed by different principles. In addition, they demonstrate the robustness of the SPT effect in AD patients, who were able to improve memory performance in the SPT condition not only with highly familiar actions but also with less familiar actions. PMID- 12650232 TI - Assigned versus self-set goals and their impact on the performance of brain damaged patients. AB - The purpose of this study was to investigate the role played by goal difficulty and goal origin (i.e., self-set vs. assigned) on the performance of patients with brain injuries in a simple arithmetic task. Eighty-seven patients with either cerebral vascular accidents or traumatic brain injuries were investigated. Patients were randomly assigned to one of three conditions: (1) one in which a specific, high goal was assigned, (2) one in which a "do your best" goal was given, and (3) one in which a personal goal has to be stated. The results indicate that both goal difficulty and goal origin had an influence on performance. Assigned difficult goals lead to better performance than assigned easy goals. Self-setting a goal did not increase performance to the same level than the assignment of a difficult goal. This lower performance increase with self-set goals can be explained by the fact that participants selected goals which were not as difficult as the assigned difficult goal. Self-set goals were attained by most of the patients and only 1 patient showed a high discrepancy between self-set goal and actual performance. These findings suggest that goal origin and goal difficulty are important moderators in the goal setting process. Furthermore, patients with brain injuries do not necessarily set unrealistic high goals. PMID- 12650233 TI - Attentional deficits in patients with persisting postconcussive complaints: a general deficit or specific component deficit? AB - This study aimed to examine attentional performance in patients with persisting postconcussive complaints, using a multi-componential perspective. Comparisons of attentional performances of 92 patients with postconcussive complaints and 86 normal controls were conducted using tests of sustained attention (Sustained Attention to Response Task; Digit Backward Span), selective attention (Stroop Word-Color Test; Color Trails Test), divided attention (Paced Auditory Serial Addition Test; Symbol Digit Modalities Test), and attentional control processing (Six Elements Test; Tower of Hanoi). Questionnaires on daily life inattentive behaviour were also administered to all participants and their significant others. A MANOVA indicated that patients with persisting postconcussive complaints demonstrated a general deficit in attentional performance as compared with their normal controls, F(18, 145) = 7.939, p = .005. These patterns still persisted when measures of emotional disturbance were controlled, F(18, 143) = 5.159, p = .005. Moreover, for sustained attention and selective attention, we were able to statistically control for speed and the differences remained. Future research should be conducted to better control the potential confound of speed for all the attentional components in order to differentiate any specific component deficits in these patients. PMID- 12650234 TI - Base rates of malingering and symptom exaggeration. AB - Base rates of probable malingering and symptom exaggeration are reported from a survey of the American Board of Clinical Neuropsychology membership. Estimates were based on 33,531 annual cases involved in personal injury, (n = 6,371). disability (n = 3,688), criminal (n = 1,341), or medical (n = 22,131) matters. Base rates did not differ among geographic regions or practice settings, but were related to the proportion of plaintiff versus defense referrals. Reported rates would be 2-4% higher if variance due to referral source was controlled. Twenty nine percent of personal injury, 30% of disability, 19% of criminal, and 8% of medical cases involved probable malingering and symptom exaggeration. Thirty-nine percent of mild head injury, 35% of fibromyalgia/chronic fatigue, 31% of chronic pain, 27% of neurotoxic, and 22% of electrical injury claims resulted in diagnostic impressions of probable malingering. Diagnosis was supported by multiple sources of evidence, including severity (65% of cases) or pattern (64% of cases) of cognitive impairment that was inconsistent with the condition, scores below empirical cutoffs on forced choice tests (57% of cases), discrepancies among records, self-report, and observed behavior (56%), implausible self-reported symptoms in interview (46%), implausible changes in test scores across repeated examinations (45%), and validity scales on objective personality tests (38% of cases). PMID- 12650235 TI - Subjective and objective assessment methods of mental imagery control: construct validation of self-report measures. AB - A study was conducted to examine the relationship between subjective and objective measures of mental imagery control. Eighty college undergraduates completed a battery of imagery tests and self-report measures to examine whether questionnaires that purport to measure imagery control or dynamic imagery ability (imagery of movement) would show a stronger relationship with objective measures of mental manipulation than would subjective measures that tap vividness of static imagery. Neither subjective measures of movement imagery nor subjective measures of stationary imagery showed meaningful relationships with objective measures of visuospatial manipulation. Additionally, subjective and objective imagery measures generally tended to dissociate. Basic component skills thought to be involved in mental manipulation, however, showed a much stronger relationship with the objective imagery tasks than did the self-report questionnaires. Findings suggest that subjective measures of imagery control do not tap the same cognitive processes involved in objective tests that require accurate imagery manipulation. PMID- 12650236 TI - Semantic verbal fluency of animals: a normative and predictive study in a Spanish population. AB - Semantic verbal fluency is a very sensible but rather unspecific tool for the detection of neuropsychological deficits. This test is highly influenced by socio cultural factors. Normative and predictive data for semantic verbal fluency of animals in a Spanish population are presented. The studied sample (n = 445) was stratified according to age and schooling. Statistical analysis reconfirmed a significant negative correlation (- 5.34) for age, and a significant positive correlation (5.34) for years of formal education. A predictive function for the production of names of animals during 1 min was established based on the subject's age and level of education: F(x) = 23.89 + age (- .144) + education (.39). The neuropsychological value and limitations of normative data and the predictive equation are discussed. PMID- 12650237 TI - North American Adult Reading Test: age norms, reliability, and validity. AB - The North American Adult Reading Test (NAART) is a quickly administered index that is widely used to estimate verbal intellectual ability. We have administered NAART to 351 healthy adults between 18 to 91 years of age to examine psychometric properties of the NAART and to elucidate influence of age, education and gender on NAART performance. The results showed that the NAART is a reliable and valid measure of verbal intelligence, comparable in psychometric properties to the WAIS R Vocabulary test and with equal psychometric properties in young, middle-aged and older adults. The NAART scores increase across the adult life span (approximately 4.5 points or approximately 0.5 SD) and with education (approximately 1.5 points/year of education) but they are unrelated to gender. The shorter version--the NAART35--is equally reliable and valid in predicting the WAIS-R Vocabulary. We provide norms as well as various equations for precise predictions of the NAART, the NAART35, and the WAIS-R Vocabulary scores based on age and education. Hierarchical regression analyses demonstrated that the verbal intelligence indexes are useful in predicting and interpreting performance on at least some, but not necessarily all neuropsychological tests, in addition to participants' age. PMID- 12650238 TI - Immunological relationship between the mother and the fetus. AB - The immunological relationship between the mother and the fetus is a bi directional communication determined on the one hand by fetal antigen presentation and on the other hand by recognition of and reaction to these antigens by the maternal immune system. There is evidence now that immunological recognition of pregnancy is important for the maintenance of gestation, and that inadequate recognition of fetal antigens might result in failed pregnancy. In contrast to HLA-A and -B Class I genes that are downregulated in human trophoblast cells, nonpolymorphic Class I molecules, e.g., HLA-G Class Ib, are expressed in extravillous cytotrophoblast and also in endothelial cells of fetal vessels in the chorionic villi as well as in amnion cells and amniotic fluid. The trophoblast does not induce transplantation immunity and resists NK- and CTL mediated lysis in vitro. According to our present knowledge, HLA-G presents antigens for gamma/delta T cells and at the same time defends the trophoblast from cytotoxic effector mechanisms. Since polymorphic MHC is absent from the trophoblast, presentation of fetally derived antigens is unlikely to be MHC restricted. gamma/delta T cells recognize a distinct group of ligands with a smaller receptor repertoire than alpha/beta T cells. Most gamma/delta T cells recognize unprocessed foreign antigens without MHC. In the decidua gamma/delta TCR-positive cells significantly increase in number and the majority of decidual gamma/delta T cells are in an activated form due to recognition of conserved mammalian molecules on the trophoblast. Following recognition of fetally derived antigens, the immune system reacts with the setting in of a wide range of protective mechanisms. Many observations suggest that pregnancy is associated with an altered TH1/TH2 balance. Maternal immune response is biased toward humoral immunity and away from cell-mediated immunity that could be harmful to the fetus. Cytokines of maternal origin act on placental development. On the other hand, antigen expression on the placenta determines maternal cytokine pattern. Normal human pregnancy is characterized by low peripheral NK activity, and increased NK activity seems to play a role in spontaneous abortions of unknown etiology. In early human pregnancy the majority of uterine lymphocytes are CD56(bright) granulated NK cells, which do not express CD16 or CD3. In rodents and humans, uterine NK cells are under hormonal control. In early pregnancy they are enriched at sites where fetal trophoblast infiltrates the decidua. The dynamics of the appearance of uterine NK cells suggest that one of the functions of these cells is control of placentation. Another protective mechanism operating in favor of pregnancy is progesterone-dependent immunomodulation. Due to stimulation by fetally derived antigens, pregnancy lymphocytes develop progesterone receptors and in the presence of progesterone produce a mediator (PIBF) that, through altering the cytokine balance, inhibits NK activity and exerts an antiabortive effect in mice. PMID- 12650239 TI - Female sex hormones as regulatory factors in the vaginal immune compartment. AB - Sexually transmitted diseases (STD) are now considered to be among the most common human infections. The incidence of STD is on the rise, which is partly due to frequent transmission during the asymptomatic phase of infection. The compounded cost of STD just in the United States is estimated to exceed $10 billion annually. STD are particularly prevalent in teenagers and young adults and the health problems caused by these diseases tend to be more severe and more frequent in woman than in men. Despite considerable efforts, a vaccine that provides protective immunity against sexually transmitted diseases in humans has not been developed. Nonetheless, research in animal models indicates that strong local and regional immune responses can influence the outcome of vaginal challenge with microbial pathogens. Vaginal immunity is an area of basic immunology that has received relatively little attention, but it is already clear that the mucosal and regional immunology of the vagina has unique features. The present review summarizes some of the anatomical, physiological and immunological features of the vagina and uterus that distinguish humans, non-human primates, rats and mice. These interspecies differences need to be taken into account in laboratory efforts to develop effective vaccines for STD in humans. PMID- 12650240 TI - The protective immune response against vaginal candidiasis: lessons learned from clinical studies and animal models. AB - Recurrent vulvovaginal candidiasis (RVVC) is a significant problem in women of childbearing ages and is caused by Candida albicans, a commensal organism of the intestinal and reproductive tracts. As a result of this commensalism, most healthy individuals have demonstrable Candida-specific adaptive immunity that is considered protective. In women with RVVC, a deficiency/dysfunction of this protective immunity is postulated to affect susceptibility to infection. Although cell-mediated immunity (CMI) is considered important for protection against mucosal candidal infections, little is understood about specific host defenses that are important at the vaginal mucosa. Studies to date suggest that a compartmentalized local, rather than systemic, immunity is important for defense against vaginitis. This review will summarize the current state of knowledge regarding protective host defense mechanisms against vaginal C. albicans infections both from clinical studies and animal models. From these data, hypotheses are presented for what host defense mechanisms appear important for resistance/susceptibility to vaginal infection. PMID- 12650241 TI - Geographic variations among infectious hypodermal and hematopoietic necrosis virus (IHHNV) isolates and characteristics of their infection. AB - Nucleotide sequence variations of a 2.9 kb fragment of infectious hypodermal and hematopoietic necrosis virus (IHHNV) isolated from samples of Penaeus monodon were determined and compared with an isolate from Hawaii. The infection characteristics of these isolates were examined by histology, in situ hybridization, and laboratory challenge studies with P. vannamei. Isolates of IHHNV were obtained from samples collected from the SE Asia region (the Philippines, Thailand, and Taiwan). Isolates of putative IHHNV were obtained from African samples (Tanzania, Madagascar, and Mauritius). The Philippine isolate had a very high nucleotide sequence identity (99.8%) to Hawaii IHHNV. The Thailand isolate showed a slightly lower identity (96.2%). The putative IHHNV sequences collected from Tanzania and Madagascar showed greater divergence from Hawaii IHHNV, 8.2% difference for Tanzania and 14.1% difference for Madagascar. A phylogenetic analysis showed that the Philippine IHHNV clustered with IHHNV found in the western hemisphere. This supports the theory that the Philippines was the origin of IHHNV that was first detected in Hawaii. In the laboratory infection study, both the Philippine and Thailand IHHNV were passed into P. vannamei, and the infected shrimp did not suffer any mortalities. In another laboratory infection, P. vannamei injected with a tissue homogenate of P. monodon from Madagascar, which tested positive for IHHNV by PCR, did not demonstrate IHHNV infection, suggesting that this putative IHHNV is not infectious to P. vannamei. PMID- 12650242 TI - Prevalence and effect of spawner-isolated mortality virus on the hatchery phases of Penaeus monodon and P. merguiensis in Australia. AB - Spawner-isolated mortality virus (SMV) has been associated with mortalities in broodstock of Penaeus monodon and with mid-crop mortality syndrome on grow-out farms. Epidemiological evidence suggested an association between the SMV status of broodstock and subsequent survival of their progeny, and this paper describes investigations into that association. The faeces of 909 broodstock in 9 different groups were tested by a polymerase chain reaction (PCR) for SMV and positive results were confirmed by an internal dot-blot. Seventy-seven spawners (8.5%) were positive for SMV with prevalence ranging from 0 to 24% among groups. The prevalence in spawners of P. monodon was higher (24%) than in P. merguiensis (4%). Three longitudinal studies were undertaken to compare the survival of progeny from broodstock that were positive to SMV with those that were not. Survival in hatchery tanks of progeny from SMV-positive spawners was lower than those from SMV-negative spawners with reductions of 23% (p = 0.01), 7.3% (p = 0.214) and 18.9% (p = 0.129) in the 3 studies. The conclusions were less consistent when examined during each of the later stages of growth in hatchery pools, nursery and grow-out ponds, with progeny from SMV-postive spawners sometimes having better survival rates. However, survival was better overall in progeny from SMV-negative spawners. Simple linear regression showed survival was negatively related to the proportion of postlarvae from SMV-positive spawners, with a decrease in survival of 5.6% for each 10% increase in the proportion of postlarvae coming from SMV-positive spawners (p = 0.006). Data from 38 ponds showed 6.71% of losses were due to SMV. If these losses were consistent across the entire industry, the annual loss due to SMV would have been approximately AUD 3 million in 1999/2000. PMID- 12650243 TI - Cloning and nucleotide sequence analysis of the chloramphenicol resistance gene on conjugative R plasmids from the fish pathogen Photobacterium damselae subsp. piscicida. AB - Transferable resistance to various drugs was investigated in Photobacterium damselae subsp. piscicida from Japan. Drug resistances were transferred via plasmids of 100, 50, and 40 kb. Resistance to chloramphenicol (Cmr) was transferred on plasmids of all 3 sizes. The Cmr gene (cat) was cloned from the 50 kb plasmids pPDP8511 and pPDP9106 transferred from P. damselae subsp. piscicida strains isolated in different years and places in Japan. Subcloning localized the cat to within 1.5 kb HindIII-HincII (or PstI) fragments. Nucleotide sequences of the coding and flanking region of the cat were determined as 1607 bp (HindIII HincII fragment) in pPDP8511 and 1568 bp (HindIII-PstI fragment) in pPDP9106, which corresponded with the sequence from nucleotides 40 to 1607 in pPDP8511. The nucleotide sequences identified an open reading frame (ORF) encoding 213 amino acid residues with a calculated molecular mass of about 24.8 kDa, a size consistent with the molecular mass of known cat gene products, and the ORF had maximum homology (99.5%) with a Type II CAT variant from Haemophilus influenzae. PMID- 12650244 TI - Epidemiology of bacterial diseases in Norwegian aquaculture--a description based on antibiotic prescription data for the ten-year period 1991 to 2000. AB - In Norway, antibacterial drugs for use in farmed fishes have to be prescribed by a veterinarian. Moreover, a national surveillance programme requires that copies of all prescriptions be sent to the Directorate of Fisheries. The prescriptions give information regarding fish farm and locality, weight and species of fish to be medicated, diagnosis, type and amount of drug prescribed, and date. These prescription data for the 10 yr period 1991 to 2000 have been recorded and systematised. A total of 6322 prescriptions issued for 11 fish species were registered; Atlantic salmon represented 87% of the prescriptions, rainbow trout 4.5%, arctic char 0.3%, turbot 3.8%, halibut 2.1%, Atlantic cod 1.2%, and European eel 0.3%. European sea-bass, wolf-fish, coalfish and wrasse represented a total of 0.4% of the prescriptions. Furunculosis was the most frequently given diagnosis in Atlantic salmon, accounting for 79% of all prescriptions for this species. Furunculosis was more frequent during the summer and early autumn, and in the western parts of Norway. Fish weighing more than 1 kg seemed to be the most susceptible. However, furunculosis has almost disappeared in Norwegian aquaculture since 1993. Vibriosis gave cause for antibiotic treatments in almost all fish species, and was the most common diagnosis in rainbow trout, halibut, turbot, cod and European eel. In Atlantic salmon, fish of small and medium size (up to 1 kg) seemed to be more at risk from vibriosis, and outbreaks were more frequent during summer, and in the western counties. Cold-water vibriosis was the second most frequently treated disease in Atlantic salmon, creating severe problems mainly in larger fish, in the northern parts of the country, and during winter and spring. The seasonal distribution was similar for winter ulcer disease, the only disease which seemed to be of increasing importance in Atlantic salmon. Non-specific diagnoses, such as 'bacterial infection' and 'fry disease', were given in a much higher proportion of prescriptions for marine fish species than in prescriptions for salmonids. PMID- 12650245 TI - Description of two new actinosporean types from a brook of Fuji Mountain, Honshu, and from Chitose River, Hokkaido, Japan. AB - Actinospore infection of oligochaetes living in the mud of 3 freshwater biotopes in Japan was studied. Using the cell-well plate method, a new aurantiactinomyxon type was found in 0.77% of the examined Tubifex tubifex oligochaete specimens from a brook near Yamanashi Prefectural Fisheries Experimental Station on Fuji Mountain. In 0.14% of Lumbriculus variagetus collected from Chitose River, near Chitose Salmon Hatchery, a new siedleckiella type was found, while at the same time 8.1% of the Lumbriculus spp. oligochaetes released triactinomyxons of Myxobolus arcticus. Of the examined Rhyacodrilus komarovi oligochaetes collected from the Mena River system, Hokkaido, 0.2, 0.6, 0.5 and 0.8% were infected with echinactinomyxon, neoactinomyxum and 2 types of triactinomyxon spores, respectively, and described in our previous paper. The oligochaetes released actinospores for several weeks. Actinospore infection showed high intensity in positive oligochaetes in the case of all the actinosporean types. Two of the actinospore types (aurantiactinomyxon and siedleckiella) presented here have not been previously described. PMID- 12650246 TI - Physical and chemical effects on viability of the Myxobolus cerebralis triactinomyxon. AB - Various chemical and physical methods for destroying the triactinomyxon (TAM) stage of the myxozoan parasite Myxobolus cerebralis were tested. The fluorescent stains propidium iodide and fluorescein diacetate were used as indicators of viability. Physical variables tested included freezing, drying, high temperature, sonication, and pressure of 6.2 x 10(7) Pa (9000 psi). Chemicals evaluated included chlorine bleach, povidone-iodine, and hydrogen peroxide. Freezing or drying for 1 h was effective in killing TAMs, but pressure was not. Temperatures above 75 degrees C for at least 5 min were also effective. Sonication with a laboratory instrument cleaner for 10 to 13 min killed and ruptured TAMs, resulting in <1.9% recovery. However, among the surviving TAMs, 39 to 58% were still viable. Chlorine concentrations of 130 ppm for 10 min were also effective at temperatures ranging from ice-water to room temperature and total hardness ranging from 10 to 500 mg l(-1). Lethal concentrations of hydrogen peroxide and povidone-iodine (10% solution) were quite high: 10% for 10 min, and 50% (5000 ppm active iodine) for 60 min, respectively. The stain results indicating TAM death were verified in 2 tests in which rainbow trout Oncorhynchus mykiss were exposed to TAMs that had been either frozen for 1 h or treated with 66 ppm chlorine as sodium hypochlorite for 1 min. None of the fish exposed to the treated TAMs became infected. These results should provide disinfection guidelines to prevent transfer of M. cerebralis TAMs to uninfected areas and provide information on the risks of parasite transfer under various treatment scenarios. PMID- 12650248 TI - Seasonal variation in white spot syndrome virus-positive samples in broodstock and post-larvae of Penaeus monodon in Thailand. AB - Records of PCR test results for white spot syndrome virus (WSSV) were reviewed in Thailand from 1998 to 2000 for wild Penaeus monodon broodstock purchased by hatcheries and for post-larvae (PL) brought by farmers to diagnostic laboratories for testing. Samples for PCR comprised DNA extracts from the last pleopod dissected from broodstock females after the first spawning and DNA extracts from whole, homogenized PL. There was a consistent pattern of fluctuation in percentage of WSSV-positive broodstock and PL. In broodstock, the fluctuation pattern was similar each year, with a low percentage (0 to 6%) from January to May and a higher percentage (6 to 18%) for the rest of the year, with a peak from September to November. The fluctuation pattern for PL was similar but offset with peaks and troughs occurring approximately 2 mo after those for the broodstock. The peak percentages of broodstock-positive samples were roughly constant from year to year, but those for PL decreased progressively in magnitude from 1998 to 2000. Examination of a small number of hatcheries in 2000 revealed that the percentage of WSSV-positive PL samples was significantly lower for hatcheries that routinely discarded WSSV-PCR-positive wild broodstock when compared to hatcheries that did not. PMID- 12650247 TI - Myxosporean plasmodial infection associated with ulcerative lesions in young-of the-year Atlantic menhaden in a tributary of the Chesapeake Bay, and possible links to Kudoa clupeidae. AB - Ulcers in Atlantic menhaden Brevoortia tyrannus (Latrobe) (Clupeidae), observed along the USA east coast, have been attributed to diverse etiologies including bacterial, fungal and, recently, harmful algal blooms. To understand the early pathogenesis of these lesions, we examined juvenile Atlantic menhaden collected during their seasonal presence in Chesapeake Bay tributaries from April to October 1999 and from March to August 2000. We conducted histopathological examinations of young-of-the-year fish from the Pocomoke River tributary, which has a history of fish mortalities and high lesion prevalence. Kudoa clupeidae (Myxozoa: Myxosporea) spores were present in the muscles of fish collected in both years. Of the fish assessed by histology in April, 5 to 14% were infected, while in May 90 to 96% were infected. Infection rates remained high during the summer. Mature spores were primarily located within myomeres and caused little or no observable pathological changes. Ultrastructure showed spores with capsulogenic cells bearing filamentous projections, and a basal crescentic nucleus with mottled nucleoplasm containing cleaved, condensed chromatin. Also, a highly invasive plasmodial stage of a myxozoan was found in the lesions of juvenile Atlantic menhaden. The plasmodia were observed in fish collected between May and July, with the maximum occurrence in late June 1999 and late May 2000. Plasmodia penetrated and surrounded muscle bundles, causing grossly observable raised lesions in 73% of all fish infected with this invasive stage. Plasmodia were also detected in the visceral organs, branchial arches, and interocular muscles of some fish. Some of the invasive extrasporogonic plasmodial lesions were associated with ulcers and chronic inflammatory infiltrates. The plasmodial stage appeared to slough out of the tissue with subsequent evidence of wound healing. Ultrastructure showed plasmodia with an elaborate irregular surface, divided into distinct ectoplasm and endoplasm; the latter contained numerous spherical vegetative nuclei, secondary generative cells, and occasional cell doublets. Our ultrastructural studies indicate that the plasmodial organisms, which are important in the etiology of the skin lesions, are myxozoans, and they may represent early stages of K. clupeidae. PMID- 12650250 TI - Further aspects of the general antimicrobial properties of pinniped skin secretions. AB - In 2 pinniped species with varying hair densities (the northern fur seal Callorhinus ursinus, and the common seal Phoca vitulina), the enzyme lysozyme and the peptide group beta-defensins are demonstrated for the first time as products of the apocrine glands and the sebaceous glands in the common integument of mammals. These antimicrobial substances are also found in the epithelial lining of the hair follicles, between the corneal lamellae of the epidermis, and in dermal histiocytes. Thus, it becomes obvious that another general defense mechanism against microbes, in addition to free sugars (Meyer et al. [2000] Aspects of general properties of skin secretions in the common seal Phoca vitulina. Dis Aquat Org 41:77-79), is active on the skin surface of haired marine mammals to which glandular lysozyme and defensins are regularly transported, so that this biotope is protected against the proliferation of potential pathogens, e.g. bacteria and fungi. PMID- 12650249 TI - Comparative susceptibility of two races of Salmo salar (Baltic Lule river and Atlantic Conon river strains) to infection with Gyrodactylus salaris. AB - The susceptibility of various races of salmonids towards infections with the skin parasitic monogenean Gyrodactylus salaris Malmberg, 1957, differs markedly. Norwegian and Scottish salmon strains are known as extremely susceptible to infection, whereas Baltic salmon races such as the Neva strain (Russian origin) and the Indals river (Swedish origin) salmon have been characterized as relatively resistant. However, the status of the many other Baltic strains has remained unknown. The present study reports on the susceptibility of the Baltic salmon from the Swedish river Lule. It was shown that this strain is susceptible to infection but to a lesser extent than the Scottish salmon. Further studies showed that injection of immuno-suppressants (dexamethasone) greatly increased population growth of G. salaris on Scottish salmon but not on the Baltic salmon. Mucous cell density on fins differed between strains, and a general trend to decreased cell density on infected fish 8 wk post-infection, compared to uninfected fish, was observed. The largest decrease in mucous cell density following infection was seen in the most resistant fish. After administration of immuno-suppressants, this decrease in mucous cell density was inhibited in the Scottish salmon but not in the Baltic salmon. Thus, there seems to be a relationship between the fishes' ability to discard mucous cells and the ability to resist infections with Gyrodactylus salaris. Although the Lule salmon seems more susceptible to infection compared to previous reports on the Neva salmon, the results support the notion that Baltic salmon strains are generally more resistant than East Atlantic salmon. PMID- 12650251 TI - Separation of polar compounds using carbon columns. AB - The objective of this review with 122 references is to provide structure and retention mechanisms of porous graphitic carbon by chromatographic analysis and computational chemical analysis of retention mechanisms. Synthesis methods of porous graphitic carbon are described. Applications for use as matrix for dynamic coating on porous graphitic carbon and direct separation of polar compounds on porous graphitic carbon demonstrated that the physical and chemical stability of graphitic carbons performed in both chromatography and extraction, especially for polar compounds, those are difficult on both silica-based and organic polymer based packing materials. The disadvantage is difficult desorption of non-polar compounds adsorbed on the surface. The development of 3.5-microm particles improves the separation power of graphitic carbon columns with the high theoretical plate number. PMID- 12650252 TI - Pressurized liquid extraction of selected molecular biomarkers in deep sea sediments used as proxies in paleoceanography. AB - Pressurized liquid extraction has been performed on a suite of deep-sea sediments to assess its capability as an extraction technique in the analysis of molecular biomarkers used in paleoceanography. Specific compounds assessed comprise long chain alkenones, n-alkanes, n-alcohols and, additionally, one diol and one keto ol. These have been extracted by both pressurized liquid extraction and ultrasonication for comparison. One key result is that the U37(K') index (based on the degree of unsaturation of the alkenones and used as a paleothermometer in paleoceanography) remains intact after both extraction techniques. In terms of biomarker concentrations, which are often used to qualitatively assess changes in marine productivity and/or terrigenous inputs, pressurized liquid extraction is substantially more efficient than ultrasonication, providing higher amounts of extracted constituents, particularly for polar compounds. PMID- 12650253 TI - Influence of the particle porosity on chromatographic band profiles. AB - The mass transfer kinetics of butyl benzoate, eluted on a monolithic RPLC column with methanol-water (65:35, v/v) as the mobile phase was investigated, using the perturbation method to acquire isotherm data and the mobile phase velocity dependence of the height equivalent to a theoretical plate of perturbation peaks to acquire kinetics data. The equilibrium isotherm of butyl benzoate is accounted for by the liquid-solid extended multilayer BET isotherm model. The total porosity of the column varies much with the butyl benzoate concentration, influencing strongly the parameters of its mass transfer kinetics and the profiles of the breakthrough curves. Using all these parameters, the general rate model of chromatography predicts band profiles and Van Deemter curves that are in excellent agreement with experimental results provided the influence of concentration on the porosity is properly taken into account. This agreement confirms the validity of the models selected for the isotherm and for the mass transfer kinetics. PMID- 12650254 TI - Monitoring dediazoniation product formation by high-performance liquid chromatography after derivatization. AB - A derivatization protocol that exploits the rapid reaction between arenediazonium ions and a suitable coupling agent followed by high-performance liquid chromatography analyses of the reaction mixture was employed to determine the product distribution, the rate constants for product formation and the association constant of 4-nitrobenzenediazonium, PNBD, ion with beta cyclodextrin, beta-CD. The derivatization of PNBD with the coupling agent leads to the formation of a stable azo dye that prevents by-side reactions of PNBD with the solvents of the mobile phase, including water, or the metallic parts of the chromatographic system that would eventually lead to erroneous identification and quantification of dediazoniation products. The results show that in the presence of beta-CD, nitrobenzene is formed at the expense of 4-nitrophenol, which is the major product in its absence. The observed rate constants for the interaction between PNBD and beta-CD increase upon increasing [beta-CD] showing a saturation profile indicative of the formation of an inclusion complex between PNBD and beta CD. By fitting the experimental data to a simplified Lineaweaver-Burk equation, the corresponding association constant and the maximum acceleration rate of beta CD towards PNBD were estimated. The protocol is applicable under a variety of experimental conditions provided that the rate of the coupling reaction is much faster than that of dediazoniation. PMID- 12650255 TI - Condensation nucleation light scattering detection with ion chromatography for direct determination of glyphosate and its metabolite in water. AB - An ion chromatography-condensation nucleation light scattering detection (IC CNLSD) method was successfully used to directly analyze glyphosate, a polar pesticide, and aminomethylphosaphonic acid, the major metabolite of glyphosate, in water without need of pre-treatment or derivatization. CNLSD gave a LOD of 53 ng/ml for glyphosate, which is much lower than the maximum contaminant level of 700 ng/ml for drinking water issued by the US Environmental Protection Agency. Spiked analytes in different matrixes were tested. A diluted commercial herbicide containing glyphosate was also evaluated. Compared to other reported methods, the IC-CNLSD method has no need of sample derivatization, pre-concentration, and mobile phase conductivity suppression. It is simple, fast and inexpensive. IC CNLSD is an ideal direct detection technique for such pesticides without chromophores or fluorophores. PMID- 12650256 TI - Identification of natural dyes in archeological Coptic textiles by liquid chromatography with diode array detection. AB - Reversed-phase HPLC with diode-array UV-Vis spectrophotometric detection has been used for identification of natural dyes in extracts from wool and silk fibres from archeological textiles. The examined objects originate from 4th to 12th Century Egypt and belong to the collection of Early Christian Art of the National Museum in Warsaw. Extraction from fibres was carried out with HCl solution containing ethanol or with warm pyridine. As the main individual chemical components of natural dyes, anthraquinone, indigoid and flavonoid dyes including alizarin, purpurin, luteolin, apigenin, carminic acid, ellagic acid, gallic acid, laccaic acids A and B and indigotin were found. For pyridine extracts another mobile phase with an optimized gradient of organic modifier concentration was used. With such an eluent the appearance of double peaks for indigotin and indirubin was eliminated. For acidic extraction of dyes from fibres, ethanol was used. Due to its higher boiling point than methanol it evaporates slower from the extraction solution enabling a more efficient extraction of dyes. PMID- 12650257 TI - Purification of quinoline yellow components using high-speed counter-current chromatography by stepwise increasing the flow-rate of the mobile phase. AB - Quinoline yellow (Color Index No. 47005) consists of multiple components that show a large difference in their partition coefficients (K), ranging from 0.03 to 3.3 in the solvent system tert.-butyl methyl ether (MTBE)-1-butanol-acetonitrile aqueous 0.1 M trifluoroacetic acid (TFA). Consequently, it requires an excessively long elution time for the simultaneous separation of all components by the standard high-speed counter-current chromatography technique, which uses a constant flow-rate of the mobile phase. In order to overcome this problem, we increased the flow-rate of the mobile phase stepwise from 0.1 to 2.0 mL/min. Using this new procedure, six components (0.2-6.1 mg) were successfully isolated from 25 mg of a commercial quinoline yellow preparation in a single run using a two-phase solvent system composed of MTBE-1-butanol-acetonitrile-aqueous 0.1 M TFA (1:3:1:5, v/v). The purified components were analyzed by high-performance liquid chromatography, electrospray ionization mass spectrometry, and nuclear magnetic resonance spectroscopy. PMID- 12650258 TI - Determination of T-2 toxin in cereal grains by liquid chromatography with fluorescence detection after immunoaffinity column clean-up and derivatization with 1-anthroylnitrile. AB - 1-Anthroylnitrile (1-AN) has been shown to be an efficient labelling reagent for the determination of T-2 toxin (T-2) by high-performance liquid chromatography (HPLC)-fluorescence detection. This reaction has been used to develop a sensitive, reproducible and accurate method for the determination of T-2 in wheat, corn, barley, oats, rice and sorghum. The method uses immunoaffinity columns containing antibodies specific for T-2 for extract clean-up, pre-column derivatization with 1-AN and HPLC with fluorescence detection for toxin determination. Ground cereal samples were extracted with methanol-water (80:20, v/v), the extracts were purified by immunoaffinity columns and the toxin was quantified by reversed-phase HPLC with fluorometric detection (excitation wavelength 381 nm, emission wavelength 470 nm) after derivatization with 1-AN. Recoveries from the different cereals spiked with T-2 at levels ranging from 0.05 to 1.5 microg/g were from 80 to 99%, with relative standard deviations of less than 6%. The limit of detection was 0.005 microg/g, based on a signal-to-noise ratio of 3:1. PMID- 12650259 TI - Effects of solvent density on retention in gas-liquid chromatography. II. Polar solutes in poly(ethylene glycol) stationary phases. AB - Effects of solvent density on the solubility of polar probes which undergo specific interactions with poly(oxyethylene) are studied. The analysis of retention data on capillary columns coated with oligomeric poly(oxyethylene) stationary phases shows that, within the experimental error, the enthalpic contribution to the solubility is practically independent of variations in the solvent density. Average values of enthalpies of solute transfer are reported for different probes and temperatures. The observed systematic decrease of solubility with the increasing density is due to a change of entropy. Some thermodynamic consequences inferred from these general results are discussed. One relevant observation is that the influence of solvent's final groups must be negligible. This is even the case for oligomers with number-average degrees of polymerization as low as 13, hosting solutes capable of strong interactions with the end hydroxyl groups of linear poly(ethylene glycols). Possible explanations for this behavior are explored through molecular dynamics simulations of the liquid solvent. PMID- 12650260 TI - Determination of carbon monoxide, methane and carbon dioxide in refinery hydrogen gases and air by gas chromatography. AB - This paper illustrates a method for determining trace amounts of CO, CH4 and CO2 with the detection limit of 0.15, 0.15 and 0.20 microg/l, respectively, in refinery hydrogen gases or in air. A simple modification of a gas chromatograph equipped with a flame-ionization detector is presented. A Porapak Q column, additionally connected with a short molecular sieve 5A packed column and a catalytic hydrogenation reactor on the Ni catalyst have been applied. The principle of the analytical method proposed is the separation of CO from O2 before the introduction of CO to the methanizer. The analytical procedure and examples of the results obtained have been presented. The modification applied makes it possible to use the GC instrument for other determinations, requiring utilization of the Porapak Q column and the flame-ionization detector. In such cases, the short molecular sieve 5A column and the methanizer can be by-passed. PMID- 12650261 TI - Development and optimization of a method for the analysis of low-molecular-mass organic acids in plants by capillary electrophoresis with indirect UV detection. AB - Capillary zone electrophoresis was developed to analyze low-molecular-mass organic acids including oxalic, tartaric, formic, malic, citric, succinic, glutaric, acetic and lactic acid. The influences of some crucial parameters such as buffer concentration, pH value, surfactant concentration and separation temperature, on electrophoretic separation were investigated. Under the conditions of 15 mM phthalate containing 0.6 mM tetradecyltrimethylammonium bromide as the run buffer (pH 5.6); separation voltage -15 kV (-263.2 V/cm) and temperature 25 degrees C, a satisfactory separation of nine organic acids was accomplished within 7 min. The detection limits (S/N=3) ranged from 0.008 to 0.08 microg/ml and the quantification limits ranged from 0.01 to 0.1 microg/ml for electrokinetic injection. The method was successfully applied to analyze organic acids in different parts of Var splendens (an edible vegetable in China). The recoveries of organic acids in real samples ranged from 88 to 121%. PMID- 12650262 TI - Determination of pyridine and adenine nucleotide metabolites in Bacillus subtilis cell extract by sweeping borate complexation capillary electrophoresis. AB - With a growing interest in new areas of bioanalytical research such as metabolome analysis, the development of sensitive capillary electrophoresis (CE) methods to analyze sub-microM concentrations of analytes in biological samples is required. In this report, the application of CE with sweeping by borate complexation is used to analyze a group of seven pyridine and adenine nucleotide metabolites derived from bacteria Bacillus subtilis cell extracts. Nanomolar (nM) detectability of analytes by CE with UV photometric detection is achieved through effective focusing of large sample plug (approximately 10% of capillary length) using sweeping by borate complexation method, reflected by a limit of detections (S/N = 3) of about 2 x 10(-8) M. Changes in metabolites concentrations were observed in cell extracts when using either glucose or malate as the carbon source in the culture medium. Concentration of pyridine and adenine nucleotides in cell extracts varied widely from 78.6 (+/-7.6) microM for nicotinamide-adenine dinucleotide in malate to 0.66 (+/-0.12) microM for nicotinamide-adenine dinucleotide phosphate in glucose culture medium. Concentrations of metabolites in a single cell were also estimated at millimolar (mM) level. The method was validated in terms of linearity, sensitivity and reproducibility. The application of CE by sweeping borate complexation allows for sensitive and reproducible analyses of nucleotide metabolites in complex biological samples such as bacteria cell extracts. PMID- 12650263 TI - Rapid and simultaneous analysis of some bioactive components in Eucommia ulmoides by capillary electrophoresis. AB - A micellar electrokinetic chromatography method was established for the qualitative and quantitative determination of three groups of bioactive components, iridoids, flavonoids and phenolic compounds, in Eucommia ulmoides. Of the eleven bioactive components being studied, ten were successfully separated in 50 mM boric acid buffer at pH 9.5, with 50 mM sodium dodecylsulfate and 4% 1 butanol, at a voltage of 20 kV, temperature of 20 degrees C and injection under high pressure at 138 kPa for 5 s in a fused-silica capillary with peak detection at 214 nm. A high reproducibility and good linearity was obtained. The relative standard deviations of the migration times in eight injections of the standards ranged from 0.64 to 1.88% and those of the corrected peak area ranged from 2.79 to 6.62%. A good linearity, with correlation coefficients in the range of 0.995 1.000, was obtained in the calibration curves of each standard from 1 to 50 ppm. The amount of these bioactive components in the bark and leaves of Eucommia ulmoides were determined. PMID- 12650264 TI - Nonaqueous based microchip separation of toxic metal ions using 2-(5-bromo-2 pyridylazo)-5-(N-propyl-N-sulfopropylamino)phenol. AB - The colorimetric metal chelating agent, 2-(5-bromo-2-pyridylazo)-5-(N-propyl-N sulfopropylamino)phenol (5-Br-PAPS), was demonstrated on a capillary electrophoresis microchip in the separation and detection of six metal ions of environmental concern, Cd2+, Pb2+, Cu2+, Co2+, Ni2+, and Hg2+. The inclusion of methanol in the buffer was found to improve both the separation efficiency and sensitivity, in addition to making the technique directly amenable to the application of solid-phase extraction. The combination of metal chelation with solid-phase extraction on a C18 silica gel microcolumn gave several hundred fold improvements in detection limits for the CE microchip measurements of toxic metal ions in water and extracted from a solid Plexiglas surface. PMID- 12650265 TI - Cerebral blood flow during acceleration in flight measured with SPECT. AB - INTRODUCTION: Positive pressure breathing for G-protection (PBG) reduces the need for fighter pilots to use the fatiguing anti-G straining maneuver (AGSM) so that they can better endure high acceleration (+Gz). The aim of the study was to determine the differences in cerebral blood flow during flight with an extended coverage anti-G suit (ECGS) with AGSM or with PBG. METHODS: Subjects were eight male members of the Finnish Air Force. Each was studied in the laboratory at +1 Gz and then during two identical flights in the back seat of the BAe Hawk Mk 51 jet trainer at +6 Gz, first with AGSM and second with PBG (24 mmHg). Regional cerebral blood flow (rCBF) was measured by injecting (99 m) Tc-ECD for deposition at +6 Gz, then scanning the subject in the laboratory an hour later using single photon emission computed tomography (SPECT). RESULT: The rCBF was 30% below baseline for both the AGSM and PBG. CONCLUSIONS: PBG maintained CBF at +6 Gz without the need for the fatiguing AGSM. PMID- 12650266 TI - Electrical stimulation prevents deterioration of the oxidative capacity of disuse atrophied muscles in rats. AB - INTRODUCTION: The purpose of this study was to evaluate the effects of electrical stimulation on disuse-atrophied muscles. METHODS: Sprague-Dawley rats were used and divided into three groups: control (C), hind-limb suspended for 7 d (HS), and HS plus transcutaneous electrical stimulation for 7 d (ES). In the ES group, transcutaneous electrical stimulation was induced at 1 Hz for 1 h every day to condition the gastrocnemius-plantaris-soleus muscles. Muscle oxidative capacity was evaluated by 31P-MRS in vivo. Maximum tension and muscle wet mass were also measured. RESULTS: Muscle oxidative capacity decreased within 1 wk in HS; however, it was maintained when electric stimulation was applied to the suspended limb. The maximum twitch tension in HS was significantly smaller than that in C (p < 0.05), while in ES it did not differ from that in C. The muscle mass was significantly smaller in the HS and ES groups compared to C (p < 0.05). CONCLUSION: These data indicated that twitch electrical stimulation was effective in preventing deterioration of muscle functions, such as maximum tension and oxidative capacity, induced by 1 wk of disuse. PMID- 12650267 TI - Effects of repeated Valsalva maneuver straining on cardiac and vasoconstrictive baroreflex responses. AB - INTRODUCTION: We hypothesized that repeated respiratory straining maneuvers (repeated SM) designed to elevate arterial BPs (arterial baroreceptor loading) would acutely increase baroreflex responses. METHODS: We tested this hypothesis by measuring cardiac baroreflex responses to carotid baroreceptor stimulation (neck pressures), and changes in heart rate and diastolic BP after reductions in BP induced by a 15-s Valsalva maneuver in 10 female and 10 male subjects at 1, 3, 6, and 24 h after performing repeated SM. Baroreflex responses were also measured in each subject at 1, 3, 6, and 24 h at the same time on a separate day without repeated SM (control) in a randomized, counter-balanced cross-over experimental design. RESULTS: There was no statistical difference in carotid-cardiac and peripheral vascular baroreflex responses measured across time following repeated SM compared with the control condition. Integrated cardiac baroreflex response (deltaHR/ deltaSBP) measured during performance of a Valsalva maneuver was increased by approximately 50% to 1.1 +/- 0.2 bpm x mm Hg(-1) at 1 h and 1.0 +/- 0.1 bpm x mm Hg(-1) at 3 h following repeated SM compared with the control condition (0.7 +/- 0.1 bpm x mm Hg(-1) at both 1 and 3 h, respectively). However, integrated cardiac baroreflex response after repeated SM returned to control levels at 6 and 24 h after training. These responses did not differ between men and women. CONCLUSIONS: Our results are consistent with the notion that arterial baroreceptor loading induced by repeated SM increased aortic, but not carotid, cardiac baroreflex responses for as long as 3 h after repeated SM. We conclude that repeated SM increases cardiac baroreflex responsiveness which may provide patients, astronauts, and high-performance aircraft pilots with protection from development of orthostatic hypotension. PMID- 12650268 TI - Motion sickness and tilts of the inertial force environment: active suspension systems vs. active passengers. AB - BACKGROUND: Maneuvering in vehicles exposes occupants to low frequency forces (< 1 Hz) which can provoke motion sickness. HYPOTHESIS: Aligning with the tilting inertial resultant (gravity + imposed horizontal acceleration: gravito-inertial force (GIF)) may reduce motion sickness when tilting is either 'active' (self initiated; Experiment 1) or 'passive' (suspension machinery; Experiment 2). METHODS: Exp 1: Twelve seated subjects were exposed to continuous horizontal translational oscillation through the body x-axis (3.1 m x S(-2) peak acceleration, 0.20 Hz) while making head tilts which were either aligned or misaligned (180 degrees out of phase) with respect to GIF. The two sessions were one week apart at the same time of day, counterbalanced for order. Head tilts were controlled by tracking a moving LED display and head trajectory was verified by accelerometry. Motion continued until moderate nausea was achieved (motion endpoint) or until a 30 min cut-off. Exp 2: A different group of 12 subjects were exposed to continuous horizontal translational sinusoidal oscillation through the body x-axis (2.0 m x S(-2) peak acceleration, 0.176 Hz) while seated in a cab which was tilted by suspension machinery around the y-axis of the ears so that GIF was aligned or misaligned (180 degrees out of phase) with the body z-axis. RESULTS: Exp 1: Mean +/- SD time to motion endpoint was significantly longer for aligned (19.2 +/- 12.0 min) than for misaligned (17.8 +/- 13.0 min; p < 0.05, two tail). Exp 2: Mean +/- SD time to motion endpoint was significantly shorter for aligned (21.8 +/- 10.9 min) than for misaligned (28.3 +/- 5.8 min; p < 0.01, two tail). CONCLUSIONS: Whether or not compensatory tilting protects against (Exp 1) or contributes to (Exp 2) motion sickness may be influenced by whether the tilting is under the active control of the person (e.g., car driver) or under external control (e.g., passenger in a high-speed tilting train). PMID- 12650269 TI - Field trials of no-decompression stop limits for diving at 3500 m. AB - INTRODUCTION: In 1990, Bogazici University (Istanbul, Turkey) launched an altitude diving program to develop techniques and safe decompression profiles for diving at high terrestrial altitudes. Following pioneering diving expeditions to lakes at high elevations in 1990-1992, it was deemed necessary to calculate new tables. METHODS: Bottom time limits for dives requiring no decompression stops (no-d) were calculated for 3500 m using linear extrapolation of U.S. Navy M values decreased by 4 ft of sea water (M4 limits). These limits were tested for 15, 18, 21, 24, 27, and 30 m of depth by diving in the Great Sea Lake at Mt Kackar (3412 m) with 10 dives per profile. RESULTS: The mean decompression sickness (DCS) risk estimated from precordial bubble scores (Spencer Scale) ranged from 0.3% to 2.8% per profile. After three expeditions, 165 dives had been achieved with a cumulative bottom time of 3199 min. No DCS occurred in dives that adhered to the M4 no-d limits. However, two cases of Type I and one case of Type II DCS were encountered where the divers accidentally exceeded those limits. DISCUSSION: Considering the estimated risk of DCS and the relatively small number of trials, a more conservative approach was used to develop a final set of high altitude dive tables. This conclusive approach used continuous compartment half lives. It is based on fitting a surface of allowable supersaturation limits using the empirical M-values from existing tables as well as our altitude diving data, together with an added constraint that forces calculated M-values to stay below the available M-value data. PMID- 12650270 TI - Self-reported headache during saturation diving. AB - INTRODUCTION: Some commercial divers have claimed that headache is a frequent symptom related to decompression following a saturation dive, but due to lack of systematic reporting there is limited knowledge of the incidence and clinical characteristics of such headaches. METHODS: During 2001, a questionnaire was distributed to divers participating in offshore saturation diving operations on the Norwegian continental shelf. Two major diving contractors participated. The survey allowed anonymous self-reporting of past and present problems with headache; pain intensity was indicated daily on a visual analog scale (VAS) from 0 to 10. Of 95 divers, 56 participated and 67 saturations were registered. RESULTS: The divers estimated a higher frequency of headaches in connection to saturation diving than in everyday life (p < 0.001). One third of the divers reported experiencing headache after they finished decompression. There was a significant increase in reports of headache on the last day of decompression (p = 0.03) and on the first day post-saturation (p < 0.001) compared with the start of decompression. Median headache duration was 6 h (range 1-84 h) and median pain score estimated on a VAS was 2.5 (range 0.1-7.8), equivalent to moderate intensity. CONCLUSIONS: Headache incidence is greater in divers during saturation diving than in everyday life. The increase is correlated to the last phase of decompression and the post-saturation period. No specific cause(s) of the headache could be identified, but we discuss possible explanations. PMID- 12650271 TI - Effectiveness of revised fluid replacement guidelines for military training in hot weather. AB - BACKGROUND: This study compared the revised U.S. Army fluid replacement guidelines (REV) with the old guidelines (OLD) on daily changes in serum sodium concentration (Na+) and body mass (BM) during Basic Combat Training at Fort Benning, GA during two successive summers. METHODS: Recruits (n = 550; OLD = 277, REV = 273) were evaluated before and after 8-12 h of outdoor military combat training in hot weather. The WBGT (mean +/- SD) averaged 26.6 +/- 1.7 degrees C for OLD and 27.4 +/- 0.9 degrees C for REV (NS). RESULTS: Serum Na+ decreased from 137.5 +/- 1.6 mEq x L(-1) to 137.0 +/- 2.1 mEq x L(-1) after outdoor military training in OLD (p < 0.05). Twenty-two recruits (8%) had serum sodium fall to below 135 mEq x L(-1) during OLD. Serum Na+ increased from 139.0 +/- 1.7 mEq x L(-1) to 139.4 +/- 2.1 mEq x L(-1) after outdoor military training in REV (p < 0.05). Only two recruits (1%) had serum Na+ fall to below 135 mEq x L(-1) during REV. BM increased an average of 1.3 +/- 1.4 kg (p < 0.05) in OLD and an average of 0.4 +/- 1.7 kg in REV (p < 0.05). CONCLUSIONS: The revised guidelines effectively reversed the decrease in serum sodium, reduced the increase in body mass, maintained hydration and minimized overdrinking during hot weather military training compared with the old fluid replacement guidelines. PMID- 12650272 TI - Acute effects of nicotine withdrawal syndrome in pilots during flight. AB - BACKGROUND: Pilots who smoke are occasionally obliged to abstain from nicotine intake during flight and may during this period exhibit certain symptoms leading to performance decrement. METHODS: We studied 20 healthy male aviators, who were regular smokers, (mean age 33.7 +/- 1.4 yr) operating military fixed- and rotary wing aircraft (C-47 Dakota, F-16, A. Bell 205). All pilots were subjected to a 12 h abstinence from cigarette smoking, during which time they performed flight duties. After landing, we studied the intensity of the nicotine withdrawal syndrome, as well as its effect on physiological parameters, psychological functions, and cognitive tasks. This was achieved by the completion of a questionnaire, measurement of BP and heart rate, and the execution of certain computerized performance assessment tests. These tests measure mental arithmetic, visual vigilance, and image free-recall. In a subsequent flight performed under similar conditions, every pilot repeated the procedure without smoking deprivation. Thus, each subject served as his own control. The Wilcoxon non parametric test was applied for statistical analysis. RESULTS: The most frequent symptoms reported during nicotine deprivation were nervousness, craving for tobacco, tension-anxiety, fatigue, difficulty in concentration, decrease in alertness, disorders of fine adjustments, prolonged reaction times, anger irritability, drowsiness, increase in appetite, and impairment of judgement. Systolic BP and heart rate tended to decrease and diastolic BP tended to rise during withdrawal, although the differences were not statistically significant. Finally, all tests recorded an impairment of cognitive functions during abstinence. CONCLUSION: Abrupt cessation of smoking may be detrimental to flight safety and the smoking withdrawal syndrome may influence flying parameters. PMID- 12650273 TI - Characteristics and toxicological processing of postmortem pilot specimens* from fatal civil aviation accidents. AB - INTRODUCTION: Autopsied biosamples from civil aviation accident pilot fatalities are submitted to the Civil Aerospace Medical Institute (CAMI) for toxicological evaluation. However, such evaluation is dependent on types and amounts of submitted samples, and obtaining suitable samples is governed by the nature of the accident. Characteristics of those samples and associated toxicological processing have not been well documented in the literature. METHOD: Therefore, the CAMI Toxicology Database was searched for these aspects. RESULTS: CAMI received samples from the pilot fatalities (CAMI cases) of approximately 80% of the 1990-2000 aviation accidents reported by the National Transportation Safety Board. Accidents and cases during June-September were higher than the other months, and more than half of the received cases had multiple samples in sufficient amounts. For example, out of 1891 cases processed for the 1996-2000 accidents, 1211 had at least adequate amounts of blood, urine, and/or vitreous humor; 324 had inadequate amounts of blood and urine; and 356 had no blood or urine. Muscle, liver, lung, and/or kidney samples were submitted in 90% of the cases, while cerebrospinal fluids were submitted in only 8% of the cases. The toxicologically preferred samples, blood and urine, were available in 78% and 56% of the 1891 cases, respectively. Out of 51 cases containing only one sample type, 46 had muscle and the remaining 5 had other sample types. Samples were primarily analyzed for combustion gases, alcohol/volatiles, and drugs. Generally, the presence of analytes is demonstrated in at least two different sample types by using two different analytical techniques for reporting a particular case as "positive." An effective quality-assurance/quality-control is maintained throughout the process. CONCLUSION: In the majority of the aviation accidents, sufficient amounts and types of biological samples were submitted for toxicological evaluation. PMID- 12650274 TI - Health of U.S. Navy submarine crew during periods of isolation. AB - BACKGROUND: An essential element in planning for long-term space missions is prediction of the medical support required. Medical data for analogous populations serving in isolated and/or contained environments are useful in predicting health risks for astronauts. METHODS: This study evaluated the rates of health events that occurred among a highly screened, healthy military population during periods of isolation using a centralized database of medical encounter records from U.S. Navy submarines. The study population was composed of U.S. Navy officers and enlisted men deployed on 240 submarine patrols between 1 January 1997 and 30 September 2000. RESULTS: A total of 1389 officers and 11,952 enlisted crew members served aboard participating submarines for 215,086 and 1,955,521 person-days at sea, respectively, during the study period. Officers had 214 initial visits to medical staff with 79 re-visits for the same condition during these patrols, while enlisted men had 3345 initial visits and 1549 re visits. Among officers, the most common category of medical events was respiratory illnesses (primarily upper respiratory infections), followed by injury, musculoskeletal conditions, infectious diseases, symptoms and ill-defined conditions, and skin problems. Among enlisted men, the most common category of medical events was injury, followed by respiratory illnesses (upper respiratory infections), skin problems, symptoms and ill-defined conditions, digestive disorders, infectious conditions, sensory organ problems (ear infections and eye problems), and musculoskeletal conditions. CONCLUSIONS: Potential mission impacting medical events reported were rare, i.e., among a crew of seven officers, only one medical event would be expected to occur during a 6-mo mission and result in 3/4 d or less of limited or no duty. Among a crew of seven enlisted men, about two medical events would be expected during a 6-mo mission and result in about 1 d of limited or no duty per medical event. PMID- 12650275 TI - Group dynamics and catecholamines during long-duration confinement in an isolated environment. AB - INTRODUCTION: The objectives of this study were to investigate possible relationships between catecholamine excretion and long-duration confinement in an isolated environment. METHODS: Stays of long duration were made by Group I (n = 4, all Russian, weeks 1-34), Group II (n = 4, mixed nationality, weeks 3-18), and Group III (n = 4, mixed nationality, weeks 22-38); other groups joined the residents for 1-wk intervals at weeks #13, #19, and #33. Data were collected from Groups I and III. RESULTS: In both Group I and Group III, the daily epinephrine excretion was significantly elevated during and after confinement compared with the pre-isolation baseline (p < 0.05), but remained mostly within normal limits during the experiment. During isolation, epinephrine excretion was significantly higher, compared with other weeks in isolation, during weeks #19 and #27 for Group I, and during week #30 for Group III. In both Group I and Group II, norepinephrine excretion increased significantly during and after isolation (p < 0.05) and was above the normal range. The daily norepinephrine excretion was significantly higher (p < 0.05) in Group I during weeks #12, #13, and #27, and during week #30 for Group III. DISCUSSION: Epinephrine excretion generally remained in the normal range. However, occasional elevations occurred due to psychological stress, which apparently correlate with changes in group dynamics. Norepinephrine excretion was above the normal range and was correlated with social events. These results suggest that to ensure optimum crew performance, entire crews along with their visiting crews should be selected collectively, rather than individually. PMID- 12650276 TI - Cognitive coping and anxiety symptoms among people who seek help for fear of flying. AB - INTRODUCTION: Fear of flying among potential passengers is rather common. In order to treat fear of flying most efficiently, it is important to find out which aspects are related to flight anxiety. The objective of the present study was to examine the extent to which various cognitive coping strategies in response to a flight were used by airline passengers and their relationship with anxiety symptoms. METHOD: A total of 261 persons who were all seeking treatment for fear of flying filled in the Cognitive Emotion Regulation Questionnaire, the anxiety subscales of the Symptom Check List (SCL-90), the Flight Anxiety Situations questionnaire, and the Flight Anxiety Modality questionnaire. RESULTS: Respondents reported using refocus on planning, rumination, putting into perspective to the highest extent and catastrophizing and other-blame to the lowest extent. Furthermore, respondents who reported using self-blame, acceptance, rumination, and/or catastrophizing to a higher extent also reported significantly higher levels of anxiety. DISCUSSION: As the present study suggests that several cognitive coping strategies are related to anxiety, intervention programs should pay attention to these aspects. The present study gives important clues about which cognitive coping strategies should be challenged in treatment of flight anxiety. PMID- 12650277 TI - Elevated bilirubin and visual perception in pilots. AB - BACKGROUND: A fatal aircraft accident of a Bo-105 helicopter on May 20, 1996 led to the suspicion that the pilot's raised serum bilirubin level could have contributed to the accident. It was suspected that an increase of bilirubin beyond 20.52 micromol x L(-1) could disturb visual perception. METHODS: In 1998 and 1999, 101 pilots with functional hyperbilirubinemia, diagnosed as Gilbert's syndrome, were subjected to perceptual-psychological testing (signal detection test) during their periodic medical examinations at the German Air Force Institute of Aviation Medicine. The results of the psychophysiological testing were compared with a control group. RESULTS: Reaction time and perception range increased with age and were independent of serum bilirubin level. CONCLUSION: There is no evidence that raised serum bilirubin levels can diminish visual perceptional performance. PMID- 12650279 TI - Short-term vestibular responses to repeated rotations in pilots. AB - INTRODUCTION: [corrected] It has been suggested that vestibular function in pilots differs from that of non-pilots. METHODS: Short-term vestibular responses to repeated rotations were explored in pilots and non-pilots using a rotating chair. Vestibulo-ocular reflex tests were done in 30 pilots (VOR-P) and 30 non pilots (VOR-NP) at rotation frequencies of 0.01, 0.02,0.04, 0.08, 0.16, and 0.32 Hz. In a separate experiment, VOR tests were done at 0.16 Hz before and after four successive velocity-step tests in twenty-five pilots (STEP-P) and their results were compared with the results of twenty-five non-pilots (STEP-NP) who were studied earlier. RESULTS: VOR-P exhibited normal VOR gains (mean values 0.47, 0.57, 0.63, 0.69, 0.67, and 0.74, respectively, for the frequencies listed above) compared with those of VOR-NP (0.48, 0.54, 0.59, 0.67, 0.72, and 0.79). VOR gain at 0.16 Hz showed little change in STEP-P (0.64 +/- 0.04 to 0.58 +/- 0.032), while it increased in STEP-NP (0.59 +/- 0.03 to 0.78 +/- 0.06). DISCUSSION: VOR gain may be a useful measurement for differentiating pilots' vestibular function from that of non-pilots. After-rotary VOR may be suppressed in pilots. PMID- 12650278 TI - Hypothermia and local cold injuries in combat and non-combat situations--the Israeli experience. AB - INTRODUCTION: Cold weather has been recognized in the Israel Defense Forces (IDF) as a potential medical and operational threat to the soldier. Although regulations have been issued to cope with this situation, every year about 20 cases of hypothermia (T(core) < 35 degrees C) and peripheral cold injuries are reported. METHODS: This study was aimed at following cold weather injuries (CI) in the IDF in the period 1994-2001. 136 cases were reported to our institute during this period. All patients were from the general population of young (20 +/ 2 yr), male soldiers in the IDF. All were classified a priori as healthy, active subjects. RESULTS: Of these patients, 51% were diagnosed with mild hypothermia and 49% with peripheral CI. Among those soldiers who suffered from peripheral CI, less than 5% were diagnosed with frostbite. Most of the cases (76%) occurred in the winter months; however, 10% occurred in the spring, 13% in autumn, and 2 cases (1%) were reported in the summer. The majority of all CI cases occurred during routine scheduled training (51%), and 15% occurred during routine duties. Of the cases, 34% occurred during combat operations (mainly ambushing and surveillance). DISCUSSION: The present study provides data on CI cases in an army where the awareness of the hazards involved in hostile environments is extensive, and in which detailed regulations aimed to prevent these injuries are common practice. The Israeli experience indicates that CI is preventable in most instances by following a few simple regulations and providing proper education to the soldiers and their commanding officers. PMID- 12650281 TI - Syndromic surveillance. PMID- 12650280 TI - Sleep apnea: a case report. PMID- 12650282 TI - This month in aerospace medicine history--March 2003. PMID- 12650283 TI - A critical need for biosignal interpretation. AB - Biosignal interpretation (BSI) methods can be used to integrate information from different physiological signals and patient-state variables both to enable decision making regarding patient status and therapeutic actions and to improve the monitoring of patients and their organ systems. Although BSI research has yielded promising results over the last decade, the number of BSI algorithms implemented in commercially available systems and used routinely in clinical practice remains limited. This is probably due to limits in our understanding of the related clinical problems and our knowledge about the strengths and weaknesses of various BSI methods and tools. We have to continually repeat the cycle of analysis, planning, execution, and evaluation to develop an understanding of the right BSI algorithm for the right clinical problem that can be embedded in a commercial monitoring system. Advancing from the present, more descriptive stage to such formal knowledge and understanding requires time and learning. PMID- 12650284 TI - Signal processing and physiological modeling--part 1: Surface analysis. AB - Signal processing offers a wide spectrum of theories, methods, and algorithms for addressing a variety of problems ranging from noise reduction, restoration, detection (of events or changes), spatiotemporal dynamics estimation, source localization, and pattern recognition. However, the classical assumptions (stationarity, linearity, etc.) usually do not apply in real situations. Recent advances, such as time-scale and time-frequency transforms, data fusion, long range dependence, and higher order moments, do not always provide sufficiently robust solutions. In this article, the basic properties and generic features of biomedical signals are examined using a wide range of examples. Algorithmic results are presented to show not only the potential performance but also the limitations of the processing resources at our disposal. The last section describes and discusses signal matching, scenario recognition, and data fusion. PMID- 12650285 TI - Signal processing and physiological modeling--part II: Depth model-driven analysis. AB - In this second of two articles on signal processing, we explore the coupling between modeling and signal processing and the critical importance of well-posed clinical questions or hypotheses, as well as a deep knowledge of the underlying mechanisms of such coupling. Our approach consists of building models either as open-loop simulators allowing us to analyze the influence of one or several internal variables on the observations, or as dynamic systems that can enhance our understanding of these underlying mechanisms to be identified in order to lead to an explanative interpretation. These explorations are illustrated by epileptic network and cardiac models, both derived at the macroscopic level. PMID- 12650287 TI - Monitoring approaches in general anesthesia: a survey. AB - The aim of this article is to give an overview of recent developments in the field of general anesthesia monitoring. We measure different physiological signals related to the functioning of several nervous systems. Using statistical or signal processing methods, monitors are derived and correlated to the dosage of anesthetic agents and to the status of the patient. Comparisons among existing monitors show that there is no one universal monitor applicable to all general anesthesia--each has its own characteristics and might be useful in particular clinical situations. Modeling of the underlying physiological mechanisms of the anesthesia may help for better understanding the interactions between anesthetic agents and nervous systems. Physiological-model-based general anesthesia monitoring and control can then be considered and optimized for each patient. PMID- 12650286 TI - Biomedical signal processing and modeling in cardiovascular systems. AB - This article revisits the subject of short-term heart-rate and arterial-pressure variability from the perspective of model structures that can be useful in defining signal processing algorithms. We draw a general scheme of the oscillation sources and interactions that contribute to cardiovascular control mechanisms and highlight the elements that were considered in different modeling works. The origin, superposition, and interaction of respiratory high-frequency (HF) and vasomotor low-frequency (LF) rhythms is presented as the integration of supraspinal and spinal circuits, vasomotor activity, and pressure control loops. We analyze in detail the necessity of considering all relevant interactions for the algorithms designed to estimate the baroreflex sensitivity. We also pinpoint the components of cardiorespiratory coupling in relation to the analysis of data from the acoustic quantification of the left ventricular volume. Finally, we analyze the tendency to produce complex behaviors even in extremely simplified systems involving interactions between oscillatory mechanisms. PMID- 12650288 TI - Methods for assessing adequacy of anesthesia. AB - Assessing adequacy of anesthesia requires evaluation of its components: hypnosis, analgesia, and neuromuscular transmission. In order to do this, many methods have been developed that process signals representing different modalities. Assessment of hypnosis requires cortical measures of the central nervous system (CNS); methods that assess analgesia concentrate on subcortical and spinal levels of the CNS; and neuromuscular transmission is a peripheral phenomenon. This article presents an overview of the current state of methods available for measuring each of these components. We conclude that, whereas important gains have been made in the area of assessment of hypnosis, mainly owing to the advancement of methods using EEG and auditory evoked potentials, and whereas neuromuscular transmission can be objectively monitored using motor nerve stimulation, assessment of analgesia still contains many challenges. PMID- 12650289 TI - Modeling for neuromonitoring depth of anesthesia. AB - This article reviews the various modeling techniques for neuromonitoring depth of anesthesia (DOA). Traditional techniques such as parametric, predictive, optimal, and adaptive modeling, proportional, integral, derivative (PID) modeling, together with modern techniques such as bispectral-based, artificial neural network-based, fuzzy logic, and neuro-fuzzy modeling, bring us to the current state of the art in DOA neuromonitoring. This article reviews historical information about each of the modern techniques and provides an example demonstrating its implementation; reviews drug pharmacokinetic/pharmacodynamic (PK/PD) and drug interaction PK/PD modeling techniques for a balanced total intravenous anesthesia (TIVA) administration; and discusses the existing technical problems and clinical challenges, suggesting new techniques necessary for the future development of a DOA monitoring and control system. PMID- 12650290 TI - Time-frequency and time-varying analysis for assessing the dynamic responses of cardiovascular control. AB - Time-frequency or time-variant methods have been extensively applied in the study of the heart-rate variability (HRV) signal. In fact, the frequency content of HRV signal has a strong correlation with the control system assessing heart rate. In particular, the power related to the low-frequency (LF) and high-frequency (HF) components have been demonstrated to correlate to the action of sympathetic and parasympathetic branches of the autonomic nervous system. However, the analysis is restricted to stationary conditions, unless time-frequency methods are employed for detecting dynamic changes that may occur during physiological and pathological conditions. This article reviews the most diffused tools for time frequency analysis, starting from linear decomposition of the signal (including short-time Fourier transform and wavelet and wavelet packet decomposition), to quadratic time-frequency distributions (including Wigner-Ville transform and Cohen's class of distributions), and finally to adaptive or time-variant autoregressive (AR) models, in both the mono- and bivariate forms. In the past few years, these approaches have been applied in several studies related to cardiovascular responses during nonstationary pathophysiological events. Among them, we will recall and discuss myocardial ischemia (spontaneous or induced), drug infusion, rest-tilt maneuver and syncope, neurophysiological, and sleep investigations. PMID- 12650291 TI - Serum ferritin and transferrin receptor concentrations during the transition from adolescence to adulthood in a healthy Swedish population. AB - AIM: As part of a longitudinal nutrition study of adolescents, changes in serum ferritin (s-Ft), serum transferrin receptors (s-TfR) and the TfR/Ft ratio from 15 to 21 y of age were studied in randomly selected Swedish adolescents. METHODS: Blood samples from 60 males and 66 females were drawn at 15, 17 and 21 y of age. RESULTS: In males, median s-Ft increased significantly from 33 microg l(-1) at 15 y to 96 microg l(-1) at 21 y, and in females, after a non-significant decrease at 17 y, from 27 at 15 y to 34 microg l(-1) at 21 y. Between 15 and 17 y, s-TfR increased significantly, in males from 5.9 to 7.7 microg l(-1), and in females from 5.2 to 7.6 microg l(-1), whereas s-TfR decreased significantly in both genders between 17 and 21 y. The log (TfR/Ft) ratio did not change in males between 15 and 17 y, but decreased significantly from 2.2 +/- 0.3 to 1.8 +/- 0.3 by the age of 21 y. In females, the TfR/Ft ratio increased significantly between 15 and 17 y, whereas a significant decrease was found by the age of 21 y. S-Ft <12 microg l(-1) at 15, 17 and 21 y was 3, 2 and 2% in males, and in females 18, 26 and 21%, respectively. CONCLUSION: This study shows an increase in s-Ft and a decline in s-TfR in males, indicating increased iron stores. In females, s-Ft did not increase significantly, whereas s-TfR and the TfR/Ft ratio decreased between 17 and 21 y, indicating that cellular iron needs were met. PMID- 12650292 TI - Altered bone metabolism in children infected with human immunodeficiency virus. AB - AIM: Data on bone homoeostasis of children infected with human immunodeficiency virus (HIV), at the time of the gain in bone mass, are very rare. To determine possible alterations in bone metabolism, 13 prepubertal vertically HIV-infected children were studied. METHODS: Viral load, CD4 count, interleukin-6 (IL-6), growth hormone, insulin-like growth factor-I (IGF-I), IGF binding protein-3 (IGFBP-3), acid-labile subunit (ALS), IGFBP-3 proteolysis, osteocalcin in blood and N-terminal telopeptide of type I collagen in urine were determined. Lumbar spine bone mineral density was examined by dual-energy X-ray absorptiometry. RESULTS: Low osteocalcin levels were found in all patients. Low IGF-I was found in only six children, who had low CD4 count and high IL-6 levels, with normal levels of IGFBP-3 and ALS, absent IGFBP-3 proteolysis and decreased bone mineral density, irrespective of viral load or growth. CONCLUSION: Low serum osteocalcin levels appear to be an initial warning sign of possible altered bone metabolism in HIV-infected children. However, only when the immune system becomes more seriously compromised is bone loss measurable by bone densitometry. PMID- 12650293 TI - Renal parenchymal damage in sibling vesicoureteric reflux. AB - AIM: To compare the incidence of renal damage in siblings of patients with vesicoureteric reflux (VUR) who presented with a documented history of urinary tract infection (UTI) with asymptomatic siblings who were diagnosed with reflux during a screening programme for hereditary VUR. METHODS: Medical and radiological records of the VUR patients (1990-2000) were examined for age, gender, mode of presentation, reflux grade and renal damage. RESULTS: VUR was noted in 226 siblings (352 ureters) in 107 families. Of the 119 siblings of index patients, 64 were investigated for a documented UTI and 55 with no history of UTI were detected during screening for sibling reflux. Dimercaptosuccinic acid scan revealed reflux nephropathy in 25 (26%) of the 97 renal refluxing units (RRU) of siblings who presented with a UTI and in 6 (7%) of the 89 RRU of asymptomatic siblings who underwent screening voiding cystourethrography (p=0.0006). Mild renal damage was present in 20 (21%) RRU of siblings with UTI and in 2 (2%) RRU of the screened siblings (p < 0.001). Moderate to severe renal damage was present in 5 (5%) RRU of siblings with UTI and in 4 (4%) RRU of the screened siblings (p > 0.05). CONCLUSION: This study demonstrated that the incidence of mild renal scarring was much higher in siblings who presented with UTI than in asymptomatic siblings. However, the incidence of moderate and severe renal scarring among asymptomatic siblings was comparable to that in siblings with VUR and UTI. PMID- 12650294 TI - Risk factors in the development of early technetium-99m dimercaptosuccinic acid renal scintigraphy lesions during first urinary tract infection in children. AB - AIM: To establish the variables that correlate with uptake defects in dimercaptosuccinic acid (DMSA) scintigraphy performed in the acute phase of a first episode of urinary tract infection (UTI). METHODS: A prospective observational study was conducted in a cohort of 158 consecutive children with a first episode of symptomatic UTI. The therapeutic delay time (TDT) was recorded. DMSA scintigraphy was performed in all children and voiding cystourethrography in 150 of them. RESULTS: 85% of the patients were younger than 2 y. Mean TDT was 33.5 h. The aetiological agent was Escherichia coli in 140 children. DMSA scintigraphy was normal in 81. Vesicoureteric reflux was detected in 33. After a multivariate logistic regression analysis the following variables were retained in the final model: TDT > or = 48 h, growth of bacteria other than E. coli, percentage of polymorphonuclear cells > or = 60% and C-reactive protein > or = 30 mg l(-1). CONCLUSION: TDT > or = 48 h, bacteria other than E. coli, percentage of polymorphonuclear cells > or = 60% and CRP > or = 30 mg l(-1) influence the findings detected in the DMSA scintigraphy performed in the acute phase of a first episode of UTI. PMID- 12650295 TI - Early determinants of first hospital admissions for asthma and acute bronchitis among Swedish children. AB - AIM: To assess the changing impact of prenatal and perinatal factors with increasing age on first admissions for asthma and acute bronchitis. METHODS: The Swedish Medical Birth Registry was linked to the National Hospital Discharge Registry for 214,276 Swedish children born in 1987 and 1988. The linkage comprised first admissions for acute bronchitis or asthma from 1987 to 1995. First admissions for gastroenteritis were included to distinguish between factors specific to asthma or bronchitis and non-specific factors related to an increased risk of hospital admission. RESULTS: Admissions for asthma and acute bronchitis before 2 y of age had a close dose-response relationship with maternal smoking, low gestational age, low maternal age and older siblings. Very premature children exposed to more than 9 cigarettes a day in utero ran a 10 times higher risk of hospitalization than postmature children without exposure to maternal smoking. However, the proportion of admissions attributed to maternal smoking was only 4.7% after adjustments for significant covariates. The number of significant determinants declined in children with their first admission for asthma after 2 y of age but male gender and low educational level were independent determinants in all age groups. Older siblings were related to an increased risk of first asthma admission before 2 y of age but a slightly reduced risk of first admission after 2 y of age. CONCLUSION: This study illustrates the varied epidemiology of asthma in different age groups. Maternal smoking was an important determinant for admission before 2 y of age but mainly in combination with other factors. The decline in hospital admissions in early childhood would therefore be small, even after the elimination of maternal smoking, if other significant perinatal and socioeconomic factors were unchanged. PMID- 12650296 TI - Cross-sectional survey of paranasal sinus magnetic resonance imaging findings in schoolchildren. AB - AIM: Sinus image abnormalities are common among children examined for indications other than sinus disease. The purpose of this study was to determine the frequency of sinus abnormalities among otherwise healthy children attending school. METHODS: Magnetic resonance imaging (MRI) was performed on 24 children aged 8-9 y, 18 of whom were re-examined after 6-7 mo. RESULTS: Sinus abnormalities were common. An abnormality was seen in 12 of the children [50%, 95% confidence interval (95% CI) 29-71%], and 9 (38%, 95% CI 19-59%) had abnormalities in the maxillary sinuses. As many as 8 (42%, 95% CI 20-67%) of the asymptomatic children (lacking clinical symptoms or findings) had abnormalities in the maxillary or ethmoidal sinuses. The follow-up examination showed that about half of the abnormal sinus MRI findings had resolved or improved without any intervention. CONCLUSION: Abnormal sinus MRI findings are common both among otherwise healthy children attending school and in totally asymptomatic children. As incidental findings, these should be interpreted as normal and do not indicate any need for treatment in children imaged for purposes other than sinus disease. PMID- 12650297 TI - Investigation of the mechanisms of therapy-related hypoglycaemia in children with acute lymphoblastic leukaemia. AB - AIM: To determine the mechanisms of fasting hypoglycaemia occurring during maintenance therapy (MT) for childhood acute lymphoblastic leukaemia (ALL). METHODS: Thirty-five children and adolescents with ALL, aged 2.4-17.4 y, were fasted for up to 16 h during MT. Nineteen of the children developed hypoglycaemia after 11 to 16 h of fasting. Blood samples for determination of metabolic changes were taken on completion of fasting. Nineteen patients underwent a glucagon stimulation test after 4 to 16 h of fasting during MT. Erythrocyte concentrations of the metabolites of methotrexate (E-MTX) and 6-mercaptopurine (E-TGN) were measured at the time of fasting. Fifteen out of 19 patients who became hypoglycaemic were re-studied 3 to 4 mo after cessation of therapy. RESULTS: In the hypoglycaemia group, plasma levels of gluconeogenic amino acids alanine and glutamine were lower (medians 117 vs 190 micromol L(-1), p = 0.009, and medians 396 vs 448 micromol L(-1), p = 0.031, respectively) than in the normoglycaemia group. Serum levels of free carnitine were lower (medians 20.3 vs 29.8 micromol L(-1), p = 0.027), free fatty acids higher (medians 3.09 vs 1.23 mmol L(-1), p < 0.001) and marked dicarboxylic aciduria was more common in the patients with hypoglycaemia (in 14/16 vs in 2/14, p < 0.001). Impaired responses to glucagon stimulation occurred in 36% (4/11) in the hypoglycaemia group and in 12.5% (1/8) in the normoglycaemia group (p = 0.243). No significant differences were detected in E-MTX and E-TGN between the groups. Most of the metabolic abnormalities returned to normal after cessation of chemotherapy. CONCLUSIONS: Low levels of gluconeogenic amino acids, especially of alanine, are associated with hypoglycaemia. Reduced hepatic glycogen stores may also be involved in the aetiology. PMID- 12650298 TI - One hundred cases of recurrent abdominal pain in children: diagnostic procedures and criteria for a psychosomatic diagnosis. AB - AIMS: To highlight the diagnostic difficulties involved in elucidating the aetiology of recurrent abdominal pain and to evaluate the use of special premises and criteria for the diagnosis of psychosomatic pain. METHODS: The study included 100 consecutive cases of recurrent abdominal pain in 6 to 15 y-old children who were evaluated at a special paediatric clinic for recurrent abdominal pain and who were followed for at least one year. The investigation included a semi structured questionnaire, laboratory investigations based on clinical data and diagnoses based on defined criteria. Three premises and seven criteria, based on the assumption that chronic negative stress can cause recurrent pain, were used for the diagnosis of psychosomatic pain. RESULTS: When the presented premises and criteria were employed for the diagnosis of psychosomatic pain, nearly half of the cases (48%) of recurrent abdominal pain could be explained. An organic or possibly organic diagnosis was found in 25% of the cases. Fourteen children had a possible psychosomatic diagnosis and in 9 cases no known diagnosis could be reached. CONCLUSION: The presented premises and criteria for the diagnosis of psychosomatic pain can be helpful when diagnosing recurrent abdominal pain, but further validation is needed. According to these criteria, a psychosomatic diagnosis was common in this series, accounting for nearly half of the cases. A medical diagnosis was evident in a quarter of the cases. PMID- 12650299 TI - Unpowered scooter injuries in children. AB - AIM: An increasing number of scooter-related injuries is being seen in emergency department. This study was initiated to establish the demographic profile and injury characteristics of the children involved, to examine the circumstances and to suggest preventive measures. METHODS: During this descriptive cohort study, data from year 2000 were collected from all children below 16 y of age who were seen in the emergency department of a Swiss hospital following an accident with an unpowered scooter. Between January and September, information regarding age, gender, injury sustained and medical management was reviewed. From October to December, data were collected prospectively. A questionnaire was sent (January September) or handed out personally (October-December) to the patients, to gather information on the circumstances surrounding the injury and the use of safety equipment. RESULTS: Thirty-six children (15M, 21F) were included in the study. The mean age was 8.8 y (range 3.0-15.8 y). The most common injuries involved the face (including six children with dental injuries) (44%) and the head (19%). Thirteen children (36%) (6M, 7F) sustained a fracture. The majority of injuries resulted from falls forward or to the side, caused by a loss of control or collision with an obstacle. Only one patient used any safety equipment. CONCLUSION: Accidents with unpowered scooters can produce severe injuries in children. Although the majority of patients sustained minor injuries (e.g. lacerations, contusions), more serious injuries can occur. The head and face were the most vulnerable. The use of protective gear, especially helmets, may reduce the number and severity of injuries. Parents, doctors and healthcare workers should be aware of the causes and prevention of injuries caused by this rapidly growing recreational activity in childhood. PMID- 12650300 TI - Wolfram (DIDMOAD) syndrome: a multidisciplinary clinical study in nine Turkish patients and review of the literature. AB - AIM: To study Wolfram syndrome (WFS) with multidisciplinary consultations and compare the results with the literature. METHODS: Nine patients fulfilled the ascertainment criteria of WFS (insulin-dependent diabetes mellitus and optic atrophy). All patients were evaluated by the departments of paediatrics, ophthalmology, audiology, urology and medical biology. RESULTS: The earliest manifestation of WFS was insulin-dependent diabetes mellitus (at a median age of 6.9 y), followed by optic atrophy (8.9 y), diabetes insipidus (10.2 y) and deafness (10.5 y). Short stature was found in five cases, delayed puberty in two cases and hypergonadotropic hypogonadism in one case. Audiography disclosed hearing loss at high frequency in all patients (100%), but only five patients had clinical subjective hearing problems. Intravenous pyelography revealed hydroureteronephrosis in eight patients. Urodynamics revealed a normal bladder in only one patient. Three patients had a low-capacity, low-compliance bladder, detrusor external sphincteric dyssynergia and emptying problem, while five had an atonic bladder. Ocular findings were optic atrophy, low visual acuity and colour vision defects. Visual field tests revealed concentric and/or peripheral diminution in five patients. Visual evoked potentials were abnormal (reduced amplitude to both flash and pattern stimulation) in seven patients. Cranial magnetic resonance imaging showed mild or moderate atrophy of the optic nerves, chiasm, cerebellum, basal ganglia and brainstem in six patients; there was a partially empty sella in one case. There was no evidence of mitochondrial tRNA(Leu) (UUR) A to G (nucleotide 3243) mutation. CONCLUSION: Wolfram syndrome should be evaluated in a multidisciplinary manner. Some specific and dynamic tests are necessary to make a more precise estimate of the prevalence and median age of the components of WFS. Short stature is a common feature in WFS. Hypogonadism may be hypogonadotropic or hypergonadotropic. Bladder dysfunction does not always present as a large atonic bladder in WFS. A low-capacity, high pressure bladder with sphincteric dyssynergia is also common. PMID- 12650301 TI - Stress and lack of social support as risk factors for small-for-gestational-age birth. AB - AIM: To determine the contributions of social support and perceived stress to the risk of small-for-gestational-age birth. METHODS: The investigation was a case control study of mothers of infants born at 37 or more completed weeks of gestation. Cases weighed less than the sex-specific 10th percentile for gestational age at birth (small for gestational age (SGA), n = 836), and controls (appropriate for gestational age (AGA), n = 870) comprised a random selection of heavier babies. RESULTS: In univariate analyses measures of informal social support, but not perceived stress or formal social support, were associated with SGA birth. It was found that Asian mothers are less likely to receive support from families and friends. After adjustment for ethnicity, informal social support was not associated with SGA. CONCLUSIONS: Support appears to reduce the risk of SGA births, but after adjustment for ethnicity this is no longer the case. Stress during pregnancy was not associated with SGA birth. PMID- 12650302 TI - Nitric oxide treatment and acute pulmonary inflammatory response in very premature infants with intractable respiratory failure shortly after birth. AB - AIM: Premature infants with respiratory failure and early-onset pneumonia have low inducible nitric oxide synthase (NOS2) and no evidence of nitric oxide (NO) toxicity. However, inhalation of NO may not be indicated in sepsis because excessive NO generation has been reported. This prospective study was designed to test the hypothesis that inhaled NO is effective in a select group of small premature infants and that the responsiveness to NO is associated with low NOS2 enzyme. METHODS: 246 very low birthweight infants (birthweight <1500 g, VLBW) were screened for severe, intractable respiratory failure (oxygenation index >40, arterial-alveolar ratio for oxygen tension <0.10) that does not respond to two doses of surfactant within 5 h from birth. Infants with severe cardiac failure or a bleeding disorder were excluded. Five of the nine eligible cases received inhaled NO. They all had prolonged rupture of foetal membranes, early-onset pneumonia and persistent pulmonary hypertension. RESULTS: All five responded strikingly, survived and appeared normal in follow-up. Airway specimens during the first day of life revealed very low NOS2, interleukin-1beta and surfactant protein A, compared with VLBW infants who had no acute infection despite histological chorioamnionitis. In early-onset pneumonia, NOS2 and other inflammatory mediators increased first during the recovery 1-2 d after birth. CONCLUSION: VLBW infants with progressive respiratory failure and infection at birth have deficient pulmonary NOS2 and cytokine response. After surfactant therapy, these infants responded strikingly to inhaled NO. An acute pulmonary inflammatory response may contribute to respiratory adaptation in early-onset pneumonia. PMID- 12650303 TI - Nasal continuous positive airway pressure versus nasal intermittent positive pressure ventilation for preterm neonates: a systematic review and meta-analysis. AB - AIM: To determine whether nasal intermittent positive pressure ventilation (NIPPV) is more effective in preterm infants than nasal continuous positive airway pressure (NCPAP) in reducing the rate of extubation failure following mechanical ventilation, and reducing the frequency of apnoea of prematurity and subsequent need for endotracheal intubation. METHODS: Randomized trials of NIPPV versus NCPAP were sought and their data extracted and analysed independently by the authors using the methodology of the Cochrane Collaboration. The analysis used relative risk (RR), risk difference (RD) and number needed to treat (NNT) with 95% confidence intervals. RESULTS: The three studies identified, comparing NIPPV with NCPAP in the postextubation period, all used synchronized NIPPV (SNIPPV), which was more effective than NCPAP in preventing failure of extubation [RR 0.21 (0.10, 0.45), RD -0.32 (-0.45, -0.20), NNT 3 (2, 5)]. Two studies compared NIPPV versus NCPAP for the treatment of apnoea of prematurity. Although meta-analysis was not possible one trial showed a reduction in apnoea frequency with NIPPV and the other a trend favouring NIPPV. CONCLUSION: SNIPPV is an effective method of augmenting the beneficial effects of NCPAP in preterm infants in the postextubation period. Further research is required to delineate the role of NIPPV in the management of apnoea of prematurity. PMID- 12650304 TI - Role of plasma and urinary calcium and phosphorus measurements in early detection of phosphorus deficiency in very low birthweight infants. AB - AIM: To analyse the role of serum and urinary calcium and phosphorus levels in early detection of mineral deficiency in very low birthweight (VLBW) infants born appropriate (AGA) and small for gestational age (SGA). METHODS: 64 VLBW infants were included in a cohort study and divided into two groups: AGA (n = 30) and SGA infants (n = 34). Then, they were divided according to the presence of radiological signs of metabolic bone disease (MBD): with MBD (n = 21) and without MBD (n = 34). Blood samples and 6 h urine collections were obtained for calcium, phosphorus, alkaline phosphatase activity and creatinine determinations between 3 and 5 wk of life. RESULTS: There were no biochemical differences between AGA and SGA. Higher values of urinary calcium (MBD = 31.9 +/- 20.2, without MBD = 19.8 +/ 15.4; p = 0.017), calciuria (MBD = 2.3 +/- 0.3, without MBD = 1.4 +/- 0.8; p = 0.037) and alkaline phosphatase activity (MBD = 369 +/- 114, without MBD = 310 +/ 93; p = 0.04) were found in infants who developed MBD. Both groups showed high tubular phosphorus reabsorption indicating mineral deficiency. CONCLUSION: Serum calcium and phosphorus levels are not good markers in early detection of mineral deficiency. However, the monitoring of calcium urinary levels may be helpful in early detection of mineral deficiency. PMID- 12650305 TI - Effect of maternal antibiotic treatment on fetal periventricular white matter cell death in a rabbit intrauterine infection model. AB - AIM: To evaluate the effects of maternal antibiotic treatment on fetal brain cell death in a rabbit intrauterine infection model. METHODS: After Escherichia coli uterine-horn inoculation in 22 pregnant rabbits, followed at various times by ceftriaxone and caesarean section, cell death in white matter (histology and fragmented DNA staining) from fetuses killed at extraction was compared across groups using the Mantel-Haenszel test and Fisher's exact test for small numbers. RESULTS: White matter cell death was consistently present at 48 h, with ceftriaxone initiation at 24 h (group 1), detectable at 84 but not 60 h, with ceftriaxone initiation at 12 h, and significantly reduced at 84 h with ceftriaxone initiation at 6 h (60% vs 100% in group 1, p < 0.001, Fisher's exact test). CONCLUSION: Early maternal antibiotic therapy delays white matter cell death in rabbit fetuses exposed to intrauterine infection. This may provide a window for preventing white matter damage. PMID- 12650306 TI - Children with nutritional rickets referred to hospitals in Copenhagen during a 10 year period. AB - AIM: To describe the prevalence of nutritional rickets among children admitted to three large paediatric departments in the Copenhagen area during a 10 y period. METHODS: Retrospective analysis of cases identified from the diagnosis registers fulfilling the diagnostic criteria for nutritional rickets. RESULTS: Forty cases were identified, distributed in two distinct age groups: 0.5-4 y (n = 31) and 9 15 y (n = 9). All cases were immigrants, of whom 95% were born in Denmark. The main symptoms in the younger age group were bowed legs and clumsy walk, and in the peripubertal group were painful joints. Two children had generalized convulsions. None of the cases had received vitamin D supplementation. CONCLUSION: Nutritional rickets is still present among immigrants in Denmark, and it is likely that the prevalence of mild cases is high. Prevention through vitamin D supplementation is important, but requires a dedicated health education effort. PMID- 12650307 TI - Influences on participation rate in a national Norwegian child development screening questionnaire study. AB - AIM: To describe the influence of demographic variables on participation rate in a child development screening questionnaire study, and to discuss the implications for data analysis and for the design of future similar studies. METHODS: Appropriate Ages and Stages Questionnaires were mailed to 2392 mothers of children aged 4-60 mo. The bivariate and multivariate influence of demographic variables on responding was investigated. RESULTS: The response rate decreased roughly linearly with the age of the child (ranging from 76% at 8 mo to 32% at 60 mo). Mother's educational level, civil status and (marginally) child's gender also contributed independently to the prediction of response. CONCLUSION: Participation rate may be interpreted in terms of interest/saliency and time demands, if a mother's interest in her baby's normative development is assumed to be great, though decreasing as the child grows. Weighting for demographic variables seems to be a viable procedure in the present project; future studies may consider measures to increase the immediacy of questionnaire completion for mothers of children aged 3 y and older. PMID- 12650308 TI - A nocturnal enuresis service for a deprived inner city. AB - AIM: Glasgow is a city where 80% of the most deprived children in Scotland live within 20% of the overall population. Of 168,000 children in Glasgow, an estimated 5000 aged 5-15 y have primary nocturnal enuresis. The aim of this study was to establish a nocturnal enuresis service for Glasgow. METHODS: Needs assessment was performed and the incident cases were documented at school entry healthcare appraisal, at the age of 5 y. A cohort of these children was followed up at 8 y of age and symptom resolution was no better than natural remission. Parents, general practitioners, clinical medical officers and a hospital paediatrician were interviewed face-to-face about their need for a service. RESULTS: Ten of 11 general practitioners expressed a need for a specialist service. Ten school nurse-led local nocturnal enuresis clinics were established. Accommodation, funding, default and managing alarms were major problems. School nurses enjoyed extra training and quickly became skilled therapists. Two research projects highlighted that most children at first attendance did not believe they could ever be dry, which predicted outcome. Deprived families, once motivated, could successfully undertake complex behavioural therapies. CONCLUSION: A comprehensive nocturnal enuresis service begins when children stop using night nappies at the age of 3-4 y. Appropriate waterproof mattress, duvet and pillow coverings should be advocated until 5 y, when desmopressin may be helpful. For continued wetting at the age of 7 y, a local service should be led by a team of well-trained nurses using enuretic alarms. PMID- 12650309 TI - Factors constraining adherence to referral advice for severely ill children managed by the Integrated Management of Childhood Illness approach in Imbabura Province, Ecuador. AB - AIM: Low referral completion rates in developing countries undermine the Integrated Management of Childhood Illness (IMCI) strategy for lowering child mortality. This study sought to identify factors constraining adherence to referral advice in a health system using the IMCI approach. METHODS: Caregivers of 160 children urgently referred to hospital were prospectively interviewed. Caregivers who accessed and did not access hospital were compared for potential referral constraining factors, including demographics, family dynamics, the severity of their child's illness, their interaction with the health system, self perceived problems, and physical and financial access. RESULTS: 67/160 (42%) referred children did not access hospital. Six factors were associated with non access, including two health worker actions: not being given a referral slip [adjusted odds ratio (OR)= 15.3, 95% confidence interval (95% CI) 4.4-64.6] and not being told to go to the hospital immediately (adjusted OR = 5.3, 95% CI 1.9 16.3). Receiving both of these interventions reduced the risk of not accessing hospital to 19%, from 96% for those who received neither intervention. Several indicators of illness severity, including caregivers' ranking of their children's illness severity, the presence of severe illness signs and mortality, were investigated and found not to be important explanatory factors. CONCLUSION: Providing a referral slip and counseling the caregivers of severely ill children to go to the hospital immediately appear to be powerful tools for increasing successful referral outcomes. PMID- 12650310 TI - Frequency of and risk factors for necrotizing enterocolitis in infants born before 33 weeks of gestation. AB - AIM: To investigate the frequency of and risk factors for necrotizing enterocolitis (NEC) among infants of <33 wk of gestation. METHODS: Prospective follow-up of 140 inpatients. RESULTS: 26 (18.6%) subjects developed NEC stage I III and 12 (8.6%) severe NEC (stage II-III). Breast milk fortifier and duration of morphine infusion emerged as the statistically significant factors associated with NEC stage I-III, but only the latter had a significant association with severe NEC. CONCLUSION: Future studies are needed to be established whether morphine is a causative factor in NEC. PMID- 12650311 TI - Higher drive for thinness in adolescent males with insulin-dependent diabetes mellitus compared with healthy controls. AB - Eating behaviour in adolescent males with insulin-dependent diabetes mellitus (IDDM) living in central Sweden was compared with that of healthy age-matched male controls using the Eating Disorder Inventory for Children and an interview. The patients were heavier than controls (p = 0.004) and had higher Drive for Thinness scores (p = 0.002). None was diagnosed as having a current eating disorder. CONCLUSION: The results of the study may indicate an increased risk of future eating disorders in males with IDDM. PMID- 12650312 TI - Benefits and risks of calcium resonium therapy in hyperkalaemic preterm infants. AB - The occurrence of an intestinal perforation in a prematurely born infant following rectal administration of a calcium resonium resin prompted a literature review to determine whether use of resins in neonates was evidence based. This revealed that the results of anecdotal series suggested that resins were useful, but the only randomized trial including prematurely born infants demonstrated that resins were less efficacious than glucose and insulin infusions. Important side effects were reported including colonic necrosis and impaction with perforation. CONCLUSION: The risks and benefits of exchange resins in prematurely born infants should be assessed in an appropriately sized randomized trial. PMID- 12650313 TI - Conversion reaction presenting as a polyradiculonevritis. AB - AIM: To show that conversion reactions should be suspected when symptomatology or instrumental checking does not conform to a precise pathological pattern in its anatomical and physiological keys. METHODS: We report the case of a 14-y-old girl who presented with signs and symptoms of Guillain-Barre polyradiculonevritis. Negative instrumental and laboratory tests led us to consider a possible diagnosis of conversion reaction. RESULTS: After a more detailed history emphasizing the close bond between the patient and her father, a poor peer relation exchange and psychological stress, we were able to identify conversion reactions through the patient's motor signs. CONCLUSION: Conversion symptoms must be suspected and searched for when the neurologic outline is difficult to clarify, and also in the case of Guillain-Barre syndrome that does not respond to plasmapheresis. PMID- 12650314 TI - Retroperitoneal abscess and omphalitis in young infants. AB - AIM: To evaluate the aetiopathogenetic factors in cases of retroperitoneal abscess in young infants, particularly the correlation with omphalitis. METHODS: We describe the cases of two infants, aged 8 and 3 wk, respectively, with a history of omphalitis during the first weeks of life and subsequent development of a retroperitoneal abscess. Both infants underwent surgical drainage of the abscess. RESULTS: In case 1, Staphylococcus aureus was found in cultures from abscess pus, and in case 2 from umbilical pus, abscess purulent material and blood. Both infants are in good health after a follow-up of 6 mo and 8 y, respectively. CONCLUSION: Retroperitoneal abscesses in young infants are usually considered to be idiopathic. A correlation with omphalitis was found in both of the reported cases and it is thought that this could have been due to an aetiopathogenetic factor. Furthermore, we stress the importance of suspicion of retroperitoneal abscesses for early diagnosis and treatment, and discuss the therapeutic strategies. PMID- 12650315 TI - Group A streptococcal osteomyelitis: severe presentation and course. AB - AIM: To evaluate the course of group A streptococcal osteomyelitis associated with severe disease nowadays. METHODS: Three consecutive cases of severe group A streptococcal disease with osteomyelitis in children that were documented in Beer Sheva, Israel are described in detail. RESULTS: Two of the three cases were postvaricella. Early in the course of the disease, the presentation resembled that of severe cellulitis. All three patients had severe osteomyelitis and required surgery, and one patient developed chronic osteomyelitis. Sepsis was diagnosed in two cases. CONCLUSION: Our cases are distinguishable from typical haematogenous staphylococcal osteomyelitis by the severe course and the extensive involvement of bone and soft tissues. The increase in severity of invasive group A streptococcal infections documented throughout the world could account for the difference between our complex cases and the previous reports. PMID- 12650316 TI - Union of Mediterranean Neonatal Societies. PMID- 12650318 TI - Total knee or uni? Benefits and limitations of the unicondylar knee prosthesis. PMID- 12650317 TI - Technetium-99m dimercaptosuccinic acid scintigraphy and pyelonephritic scarring in newborn children. PMID- 12650319 TI - Radiologic case study. Traumatic dislocation of the knee. AB - Imaging in knee dislocation requires multiple modalities and a comprehensive assessment of ligaments, cartilage, bone, nerve, and vascular anatomy. Magnetic resonance imaging is essential in the work-up of these patients and MRA is a promising modality for detecting vascular injury. PMID- 12650320 TI - Intraoperative closed reduction and percutaneus fixation of displaced femoral neck fractures with the trochanteric gamma nail. PMID- 12650321 TI - Technical aspects of medial versus lateral minimally invasive unicondylar arthroplasty. PMID- 12650322 TI - The effectiveness of autologous chondrocyte implantation for treatment of full thickness articular cartilage lesions in workers' compensation patients. AB - This prospective longitudinal analysis evaluated the effectiveness of autologous chondrocyte implantation in 24 workers' compensation patients with articular cartilage injuries >2 cm2. Mean lesion size was 4.7 cm2, (range: 2-10 cm2). Nineteen lesions were on the distal femur and 5 were on the patella. Clinical assessment scores improved from a mean of 3.2 at baseline to 6.8 one year postoperatively, with 78% rated good or excellent. Among patients with > or = 1 year follow-up, 63% returned to unrestricted work status at a mean of 7 months; an additional 22% returned to modified work. PMID- 12650323 TI - A new classification and treatment protocol for combined fractures of the femoral shaft with the proximal or distal femur with closed locked intramedullary nailing: clinical experience of 63 fractures. AB - The medical records and radiographs of 63 patients, who were admitted between 1989-1997, with a combined femur fracture, were reviewed. Associated injuries were present in 38 (60%) patients. The combined fractures were classified into four major types depending on their anatomical position: type I, femoral shaft fracture combined with hip neck fracture; type II, femoral shaft fracture combined with a trochanteric fracture; type III, femoral shaft fracture combined with a distal femur fracture; and type IV, femoral shaft fracture combined with a proximal or distal femur fracture. The fractures were treated with locked intramedullary nailing and additional free cancellous 6.5-mm screws as needed. Fifty-six fractures healed without further operations. Of the remaining 6 fractures, 2 were material failures, 1 malunion with 3-cm shortening and external rotation of the femoral diaphysis, 2 early infections of the surgical wound, and 1 pseudarthrosis of the femoral shaft. All fractures were healed between 16 and 32 weeks (average: 20 weeks). PMID- 12650324 TI - The effect of delayed autogenous bone grafting on the healing of diaphyseal defects in a canine model. AB - Bone grafting of osseous defects is often delayed to minimize the risk of infection, however, the effect of this delay on defect healing is not clear. Unilateral oblong unicortical diaphyseal femoral defects (30 x 4.5 mm) were created in 3 groups of 18 adult canines that were grafted with autogenous bone immediately, at 2 weeks, and at 6 weeks. After an 8-week healing period, paired femurs, h consisted of defect versus intact specimens, were assessed by biomechanical testing and microradiography. No differences were demonstrated biomechanically or radiographically between the healing defects with immediate versus delayed bone grafting. This study suggests that within the first 6 weeks postoperatively osseous defects can be grafted with autologous bone without adversely affecting bone graft incorporation. PMID- 12650325 TI - Lumbar pedicle morphology in adolescent idiopathic scoliosis. AB - Using computed tomography of lumbar spine pedicles, pedicle morphology was documented in 30 patients with adolescent idiopathic scoliosis. Lumbar pedicles are relatively symmetrical in patients with adolescent idiopathic scoliosis; however, a statistically significant difference was found between males and females at L2-L4. A trend for increased transverse pedicle width was observed on the concavity versus the convexity. At levels L3-L5, 90% of the pedicles were >5.5 mm but pedicles Li and 12 were smaller, only >5.5 mm in 34% and 42%, respectively. Safe insertion of 5.5-mm screws is possible at 13-15, but more cephalic pedicles necessitate caution. PMID- 12650326 TI - Interobserver variability of the Insall-Salvati ratio. AB - Thirty six patients presenting with knee pain were enrolled in a prospective study to assess the variability of the radiographic measurement of patellar height by the traditional and modified Insall-Salvati ratios. The traditional and modified Insall-Salvati ratios were measured from a lateral knee radiograph using a standard hand-held ruler by four physicians in varying order. The observers were in agreement in classifying the radiographs in two thirds of the cases When the traditional method was used but in less than one-half of the cases when the modified ratio was used. The source of the difference in the two ratios is a greater variability in measuring the distance from the inferior margin of the articular surface to the tibial tubercle. PMID- 12650327 TI - Acute displaced femoral shaft fracture in a football player. PMID- 12650328 TI - Primary skeletal muscle tuberculosis. PMID- 12650329 TI - Bilateral hip dislocation and pubic diastasis in familial nail-patella syndrome. PMID- 12650330 TI - Mycobacterium marinum osteomyelitis. PMID- 12650331 TI - Compartment syndrome of the hand caused by computed tomography contrast infiltration. PMID- 12650332 TI - Arthroplasty in the treatment of subcapital hip fracture. AB - After reviewing recent literature on the treatment of displaced intracapsular fractures of the femoral neck (Garden types III and IV) and from personal experience, a number of conclusions can be made. Orthopedic surgeons and hospitals face the challenge of providing the treatment most beneficial to patients with intracapsular, subcapital hip fractures in the most cost-effective way. The numbers of patients will increase annually and exceed the 125,000 per year at present in the United States. Most authors agree that fractures with the least displacement and younger, more demanding patients will do well with a precise fracture reduction without delay and an accurately placed internal fixation system. However, an overall median risk for reoperation 2 years after internal fixation is 35%. A patient with a displaced intracapsular fracture will need to consider monopolar, bipolar, or THA as the treatment of choice. Monopolar and bipolar arthroplasty have a reduced survivorship compared to THA and are not as suitable for the younger, more active patient. A large femoral head implant leads to decreased motion from increased friction and an undersized head implant leads to reduced contact area with increased erosion and pain. Bipolar arthroplasties, while allowing early mobilization, may develop some of the characteristics of monopolar implants if motion is not mainly at the internal joint. The increased cost may not justify their use over monopolar arthroplasty. Should dislocation occur, monopolar implants are easier to reduce (closed) than bipolar. Ceramic heads on monopolar or bipolar arthroplasties offer reduced wear and less erosion of the acetabulum. Total hip arthroplasty provides early mobilization, long-term pain relief, and little additional morbidity at surgery. The increased rate of early dislocation may be related to surgeon skill rather than an inherent failure of the system. If the early dislocators are removed from consideration, the complication rate drops to equal that of monopolar and bipolar implants. Total hip arthroplasty also is cost effective. Total hip arthroplasty may be the only option if pre-existing arthritis, significant osteoporosis, or Paget's disease of the pelvis is present. PMID- 12650333 TI - Quasi-steady state approximation to a fungal growth model. AB - In a previous paper, we proposed a fungal growth model (Lamour et al., 2001 IMA J. Math. Appl. Med. Biol., 17, 329-346), describing the colonization and decomposition of substrate, subsequent uptake of nutrients, and incorporation into fungal biomass, and performed an overall-steady-state analysis. In this paper we assume that where nutrient dynamics are much faster than the dynamics of fungal biomass and substrate, the system will reach a quasi-steady-state relatively quickly. We show how the quasi-steady-state approximation is a simplification of the full fungal growth model. We then derive an explicit fungal invasion criterion, which was not possible for the full model, and characterize parameter domains for invasion and extinction. Importantly, the fungal invasion criterion takes two forms: one for systems where carbon is limiting, another for systems where nitrogen is limiting. We focus attention on what happens in the short term immediately following the introduction of a fungus to a fungal-free system by analysing the stability of the trivial steady state, and then check numerically whether the fungus is able to persist. The derived invasion criterion was found to be valid also for the full model. Knowledge of the factors that determine invasion is essential to an understanding of fungal dynamics. The simplified model allows the invasion criterion to be tested with experimental data. PMID- 12650334 TI - Epidemics in predator-prey models: disease in the predators. AB - The author has recently proposed and investigated models for the study of interacting species subject to an additional factor, a disease spreading among one of them, that somehow affects the other one. The inadequacy of such a model comes from the basic assumption on the interacting species. It is well known that the cycles found in the Lotka-Volterra system exhibit a neutral stability, and this phenomenon is carried over to the proposed model. Here we would like to extend the study to account for population dynamics leading to carrying capacities, i.e. logistic behaviour. This corresponds to the so-called quadratic predator-prey models found in the literature. We are able to show that in some cases the trajectories are bounded, and also analyse the local stability of some equilibria. PMID- 12650335 TI - The stability of internal equilibria in predator-prey models with breeding suppression. AB - We present two models that represent the suppression of breeding by prey in response to short-term increases in predation pressure. For both of these models, we have been able to produce analytic conditions for the local stability of the interior steady state, in terms of the values of combinations of these parameters. Although our models are as simple as possible to capture the effect of breeding suppression, the expressions for local stability, even in their simplest form, are complex. Thus, we come to the important conclusion that there is no simple and general rule for the effect of the behaviours described here (anti-predatory breeding suppression and prey switching by predators) on the stability of population dynamics. Rather, effects will be system specific. However, we hope that the results and methodological framework outlined here will provide a useful tool for others to investigate the consequences for particular real systems. PMID- 12650336 TI - A non-linear model for a sexually transmitted disease with contact tracing. AB - A non-linear model is developed for an epidemic with contact tracing, and its dynamic is studied. We present the data for the Cuban HIV/AIDS epidemic and fit the non-linear model, we obtain estimates for the size of the Cuban HIV epidemic, and for the mean time for detecting a person that is infected with HIV. PMID- 12650337 TI - Seasonal dispersal patterns of Frankliniella fusca (Thysanoptera: Thripidae) and tomato spotted wilt virus occurrence in central and eastern North Carolina. AB - The seasonal abundance and temporal pattern of Frankliniella fusca Hinds dispersal were monitored from 1996 to 2000 at 12 locations in central and eastern North Carolina. The predominant vector species of tomato spotted wilt virus (TSWV) captured across all locations was F. fusca (98%). The temporal patterns of F. fusca dispersal observed during spring seasons varied among locations in all years except 2000. Regression analysis estimated that times of first flight in the spring seasons varied among locations, whereas flight duration intervals were similar. Temporal patterns of F. fusca captured varied significantly between aerial traps placed 0.1 and 1.0 m above the soil surface. Fewer total thrips were captured at 0.1 m, although thrips dispersal occurred earlier and over a greater time interval compared with 1.0-m traps. Temporal patterns of TSWV occurrence differed among locations in the spring seasons of 1999 and 2000, whereas patterns of virus occurrence were similar during the fall seasons. Patterns of F. filsca dispersal and subsequent TSWV occurrence were synchronous at locations in 1999 and 2000 where the greatest number of TSWV lesions was recorded. Knowledge of the temporal patterns of F. fiasca dispersal and TSWV occurrence may be a useful indicator for describing the time when susceptible crops are at highest risk of TSWV infection. PMID- 12650338 TI - Impact of a novel species of Nosema on the southwestern corn borer (Lepidoptera: Crambidae). AB - A study was undertaken to elucidate the impact of an undescribed Nosema sp. on the southwestern corn borer (SWCB; Diatraea grandiosella Dyar). The Nosema sp. (isolate 506) included in the study was isolated from an overwintering SWCB larva in Mississippi. It was highly infectious per os, with a median infective dose of 2.0 x 10(3) spores per larva. Even at the highest dosage tested (10(7) spores per larva), minimal mortality (< or = 3%) was observed in infected larvae, pupae, and adults reared in the laboratory on an artificial diet. However, infected pupae (0 and 7-d-old) were smaller, and the time to adult eclosion from pupation was slightly increased. Furthermore, the number of eggs produced by infected SWCB female moths substantially decreased (32%), and this effect was most pronounced on day 2, when the greatest number of eggs were oviposited by infected and noninfected moths. For eggs produced by infected females mated with infected males, hatch was slightly decreased by 16 and 15% for eggs laid on days 2 and 3, respectively. In addition, egg hatch was reduced in eggs oviposited by noninfected females mated with infected males on day 3. A low prevalence of infection (< 6%) was observed in the F1 generation originating from infected females mating with noninfected males, from noninfected females mating with infected males, and from infected females mating with infected males. Nosema 506 spores were observed in the proximity of reproductive tissues of infected female and male moths. Spores also were detected on the chorion surface and within eggs laid by infected females. Furthermore, 1-11% of larvae hatching from surface sterilized eggs were infected by Nosema 506 indicating a transovarial mechanism of transmission. PMID- 12650339 TI - Bionomics of Scymnus (Pullus) louisianae J. Chapin (Coleoptera: Coccinellidae) as a predator of the soybean aphid, Aphis glycines Matsumura (Homoptera: Aphididae). AB - A field population of Scymnus louisianae Chapin (Coccinellidae) was found attacking soybean aphids, Apis glycines Matsumura (Aphidae), a pest recently introduced into Kentucky. This coccinellid had not previously been found in Kentucky. A greenhouse population of S. louisianae was established and its predation on A. glycines studied under laboratory conditions. Total time to develop from egg to adult was about 20 d. About 70% of immatures survived to adulthood and they consumed approximately 100 aphid nymphs per beetle larva during the beetle's four larval instars. Adults lived for an average of 47 d (mated males) and 63 d (mated females) and, during their total adult lifetime, mated males consumed an average of 665 nymphs and mated females consumed 1261 nymphs. All developmental times and predation rates were comparable to those reported for other aphidophagous Scymnus spp. which, in conjunction with reports that Scymnus spp. are effective predators of cotton aphids, Aphis gossypii Glover, suggests that S. louisianae is a potentially important predator of A. glycines in the southern United States. PMID- 12650340 TI - Combining azadirachtin and Paecilomyces fumosoroseus (Deuteromycotina: Hyphomycetes) to control Bemisia argentifolii (Homoptera: Aleyrodidae). AB - Both azadirachtin and Paecilomyces fumosoroseus (Wize) Brown & Smith have been used to control the whitefly Bemisia argentifolii Bellows & Perring, but with only moderate effectiveness. Azadirachtin is a botanical insecticide derived from the neem tree, and P. fumosoroseus is an entomopathogenic fungus. To test whether these two agents might be more effective for whitefly control if used together, different rates of each were combined in laboratory bioassays in factorial treatment. Both tank mixes and separate sprays were tested. Up to 90% nymphal mortality was obtained when both the fungus and azadirachtin were combined, a significant increase over the 70%, or less, mortality obtained when only one agent was used; however, the combined effects were less than additive. Azadirachtin had moderately inhibitory effects on growth and germination of P. firmosoroseus, which may explain this antagonism. PMID- 12650341 TI - MCH released in a novel pheromone dispenser prevents spruce beetle, Dendroctonus rufipennis (Coleoptera: Scolytidae), attacks in south-central Alaska. AB - Field tests of 3-methyl-2-cyclohexen-1-one (MCH), the antiaggregation pheromone of the spruce beetle, Dendroctonus rufipennis Kirby, were conducted in south central Alaska in stands of Lutz spruce, Picea x lutzii Little, and Sitka spruce, P. sitchensis (Bong.) Carr. The deployment of MCH in a novel releaser significantly reduced trap catches and spruce beetle attacks on standing live spruce by 96 and 87%, respectively. The results of this study demonstrate the first successful field test of MCH in Alaska for the prevention of spruce beetle attacks on standing, live spruce. PMID- 12650342 TI - Lethal and sublethal effects of a neem-based insecticide on balsam fir sawfly (Hymenoptera: Diprionidae). AB - Lethal and sublethal effects of Neemix 4.5 EC, a commercial neem preparation, on balsam fir sawfly, Neodiprion abietis (Harris), were determined in the laboratory. Larval mortality of N. abietis increased in a concentration-dependent manner, and lethal time decreased with increasing Neemix 4.5 EC concentration. Fifty percent of the larvae died after 4.6 d at a concentration of 90 ppm azadirachtin (AZA) and 12.3 d at a concentration of 0.08 ppm. Neemix 4.5 EC showed some deterrent effects to feeding site selection on N. abietis larvae at high concentrations, but not at low concentrations. Strong reduction of food intake by N. abietis larvae after exposure to Neemix 4.5 EC was demonstrated by significant reduction of frass production. Larvae fed on Neemix 4.5 EC-treated foliage at 90 ppm AZA produced only 16% as much frass as that produced by larvae fed on control foliage (0 ppm). Neemix 4.5 EC at a concentration of 0.08 ppm AZA retarded larval and pupal development. Sublethal doses significantly reduced pupal weight and adult emergence, although the sex ratio of N. abietis adults was not affected. Results indicate that sublethal effects of Neemix 4.5 EC on N. abietis may contribute greatly to the overall field efficacy. PMID- 12650343 TI - Survival and development of Lymantria monacha (Lepidoptera: Lymantriidae) on North American and introduced Eurasian tree species. AB - Lymantria monacha (L.) (Lepidoptera: Lymantriidae), the nun moth, is a Eurasian pest of conifers that has potential for accidental introduction into North America. To project the potential host range of this insect if introduced into North America, survival and development of L. monacha on 26 North American and eight introduced Eurasian tree species were examined. Seven conifer species (Abies concolor, Picea abies, P. glauca, P. pungens, Pinus sylvestris with male cones, P. menziesii variety glance, and Tsuga canadensis) and six broadleaf species (Betula populifolia, Malus x domestica, Prunus serotiaa, Quercus lobata, Q. rubra, and Q. velutina) were suitable for L. monacha survival and development. Eleven of the host species tested were rated as intermediate in suitability, four conifer species (Larix occidentalis, P. nigra, P. ponderosa, P. strobus, and Pseudotsuga menziesii variety menziesii) and six broadleaf species (Carpinus caroliniana, Carya ovata, Fagus grandifolia, Populus grandidentata, Q. alba, and Tilia cordata) and the remaining 10 species tested were rated as poor (Acer rubrum, A. platanoidies, A. saccharum, F. americana, Juniperus virginiana, Larix kaempferi, Liriodendron tulipfera, Morus alba, P. taeda, and P. deltoides). The phenological state of the trees had a major impact on establishment, survival, and development of L. monacha on many of the tree species tested. Several of the deciduous tree species that are suitable for L. monacha also are suitable for L. dispar (L.) and L. mathura Moore. Establishment of L. monacha in North America would be catastrophic because of the large number of economically important tree species on which it can survive and develop, and the ability of mated females to fly and colonize new areas. PMID- 12650344 TI - Resistance to locust leafminer (Coleoptera: Chrysomelidae) in black locust. AB - Locust leafminer, Odontota dorsalis (Thunberg), activity was observed for 2 yr on 70 black locust, Robinia pseudoacacia L., trees grown from seed collected from its native range. Four black locust seedlings from Tennessee had low numbers of eggs, larvae, pupae, and mines, and were selected for clonal propagation. These clonal propagules were resistant to locust leafminers collected from three different counties in Maryland. This resistance was also observed in controlled choice and no-choice tests in outdoor screen houses. Adults showed different ovipositional preferences for black locust trees from different seed sources, which were reflected in lower numbers of larvae, pupae, and mines in resistant clones. It was concluded that antixenosis and antibiosis were involved in the mechanisms of resistance to the locust leafminer in black locust. PMID- 12650345 TI - Comparison of microcapsule density with various apple tissues and formulations of oriental fruit moth (Lepidoptera: Tortricidae) sprayable pheromone. AB - The density of microcapsules was compared on 'Golden Delicious' mature and immature foliage, fruit, and 1-yr-old limbs after dip treatments with Phase III oriental fruit moth, Grapholita molesta (Busck), microencapsulated pheromone. Microcapsules were counted with a dissecting microscope at 50x magnification after apple tissues were treated with a two-step dye staining process to increase the visibility of the microcapsules. The number of Phase III microcapsules on 1 yr limbs treated with a field concentration was two- to threefold greater than the number of microcapsules on similarly treated fruit or foliage. Phase I, III, and V OFM MEC and Checkmate OFM-F formulations of microencapsulated pheromone also were compared to determine their abundance on mature apple foliage. The Phase V OFM MEC formulation had the highest density of microcapsules when mature foliage was treated at the field rate. The Phase I treatment had the greatest difference between upper and lower leaf surfaces with 18-fold greater microcapsule density on the upper surface. On mature apple leaves treated with Phase III MEC, the number of microcapsules/cm2 was two- to threefold greater on fields of view without the mid-vein than those that included the mid-vein. The cuticle structure and abundance of trichomes are two factors that may have contributed to differences in microcapsule density among plant tissue types, top and bottom leaf surfaces, and fields of view with and without the mid-vein. PMID- 12650346 TI - Watermelon growth, fruit yield and plant survival as affected by squash bug (Hemiptera: Coreidae) feeding. AB - Adult squash bugs, Anasa tristis (De Geer) (Heteroptera: Coreidae), were confined on watermelon plants at differing phenological stages and at densities of zero to four per plant in one trial and zero to 32 per plant in three additional trials. Squash bugs were allowed to feed on the plants until plants died or fruit matured. Plant foliage, roots, and fruit were harvested and weighed to determine effects on growth and productivity. Growth and fruit production was regressed on number of squash bugs and results indicated that an increasing density of squash bugs feeding on vining and flowering stage plants resulted in significant reductions in plant growth and fruit yield. When plants were infested at the fruit set stage of growth, there was either less effect or no effect on plant growth and fruit production. Plant mortality increased as the density of squash bugs increased regardless of the stage of growth when plants were infested with squash bugs. PMID- 12650347 TI - Bt sweet corn and selective insecticides: impacts on pests and predators. AB - Sweet corn, Zea mays L., is attacked by a variety of insect pests that can cause severe losses to the producer. Current control practices are largely limited to the application of broad-spectrum insecticides that can have a substantial and deleterious impact on the natural enemy complex. Predators have been shown to provide partial control of sweet corn pests when not killed by broad-spectrum insecticides. New products that specifically target the pest species, while being relatively benign to other insects, could provide more integrated control. In field trials we found that transgenic Bt sweet corn, and the foliar insecticides indoxacarb and spinosad are all less toxic to the most abundant predators in sweet corn (Coleomegilla maculate [DeGeer], Harmonia axyridis [Pallas], and Orius insidiosus [Sav]) than the pyrethroid lambda cyhalothrin. Indoxacarb, however, was moderately toxic to coccinellids and spinosad and indoxacarb were somewhat toxic to O. insidiosus nymphs at field rates. Bt sweet corn and spinosad were able to provide control of the lepidopteran pests better than or equal to lambda cyhalothrin. The choice of insecticide material made a significant impact on survival of the pests and predators, while the frequency of application mainly affected the pests and the rate applied had little effect on either pests or predators. These results demonstrate that some of the new products available in sweet corn allow a truly integrated biological and chemical pest control program in sweet corn, making future advances in conservation, augmentation and classical biological control more feasible. PMID- 12650348 TI - Evaluating companion planting and non-host masking odors for protecting roses from the Japanese beetle (Coleoptera: Scarabaeidae). AB - Effectiveness of companion planting, and use of nonhost masking odors were evaluated under field conditions for protecting roses against the Japanese beetle, Popillia japonica Newman. Three reputedly effective companion species, rue (Ruta graveolens L.), zonal geranium (Pelargonium x hortorum Bailey), and garlic chives (Allium scheonparum L.) were interplanted with roses in replicated garden plots. Numbers of beetles on these roses were compared with rose-only control plots on 6 d during beetle flight. The masking odor hypothesis was tested by hanging mesh bags of aromatic herbs or other sources of reputedly repellent nonhost volatiles around potted roses in the field. Treatments included crushed red pepper (Capsicum frutescens L.), fennel seeds (Foeniculm vulgare Miller), crushed spearmint (Mentha picata L.), cedar shavings (Juniperus sp.), osage orange fruits (Maclura pomifera (Raif) Schneid.), and fleshy gingko seeds (Gingko biloba L.). No treatment significantly reduced numbers of beetles relative to the controls. Interplanting with geraniums significantly increased numbers of Japanese beetles on roses. Similarly, roses surrounded by sachets with fennel seeds, cedar shavings, crushed red pepper, or osage orange fruits had significantly more beetles than the control plants on two or more sample dates. Our results suggest that the use of companion or reputedly repellent plants or plant odors probably will be ineffective for protecting roses or other highly susceptible ornamentals from P. japonica. Use of such tactics in an effort to discourage other garden pests might even increase Japanese beetle damage in those plantings. PMID- 12650349 TI - Tunneling activity of subterranean termites (Isoptera: Rhinotermitidae) in sand with moisture gradients. AB - Tunnel formation by Coptotermes formosanus and Reticulitermes flavipes was studied in a two-dimensional foraging arena with a moisture gradient. Moisture did not appear to affect tunneling when termite first emerged from the central release chamber. But as termites of both species moved further away from the chamber and into a moisture gradient, they tunneled significantly (P < 0.05) more in sand with a higher moisture content than in sand with a lower moisture content. Over the 10 d test period, both termite species tunneled more in sand with a higher moisture content. Fractal analysis indicated that regardless of the sand moisture content, termite tunnel geometry had a fractal dimension and termites generally tunneled more in higher moisture sand. PMID- 12650350 TI - Effects of contaminants on bait acceptance by Solenopsis invicta (Hymenoptera: Formicidae). AB - Three commonly used fire ant baits, Amdro (0.73% hydramethylnon [AI]), Ascend (0.011% abamectins [AI]), and Maxforce (1.0% hydramethylnon [AI]), were exposed to potential, volatile contaminants. The contaminants included the insecticides Orthene Fire Ant Killer (75.0% acephate [AI] ), Cyren (44.6% chlorpyrifos [AI]), and Tempo 2 (24.3% cyfluthrin [AI]); cigarette smoke; gasoline (unleaded, 89 octane); and fertilizer (10-10-10). Fire ant baits previously exposed for 48 h to these contaminants were analyzed using gas chromatography analysis. Orthene Fire Ant Killer, Cyren, Tempo 2, cigarette smoke, and gasoline had volatile components transferred to the baits. Baits exposed to these products were used in a field evaluation of bait acceptance by Solenopisis invicta Buren, the red imported fire ant. Uncontaminated Amdro was significantly preferred by S. invicta over Amdro contaminated by smoke, Cyren, Tempo 2, and gasoline. Uncontaminated Maxforce was significantly preferred over Maxforce contaminated by Tempo 2, Cyren, and gasoline, and uncontaminated Ascend was preferred over Tempo 2- and Cyren contaminated Ascend. Orthene-exposed Amdro, Maxforce, and Ascend baits, and smoke exposed Maxforce and Ascend baits were not significantly different from the control. These results indicate that volatile insecticides and products can contaminate fire ant baits. Some insecticides and products, such as gasoline, can significantly affect bait palatability and may adversely impact control. PMID- 12650351 TI - Efficacy of bifenthrin treatment zones against red imported fire ant. AB - Exclusion of ants, particularly red imported fire ant, Solenopsis invicta (Buren), from homes, nursing facilities, hospitals, and electrical housings is an important strategy in urban and rural pest control. We conducted a laboratory bioassay to determine the repellency of granular bifenthrin (Talstar: rate 2.087 kg of formulated product/92.88 m2 or 4.6 lb formulated product/1000 feet2 or 4.2 g active ingredient/92.88 m2) to S. invicta foragers. In the field, we compared the efficacy of three widths (0.3, 2.0, and 3.0 m) of granular bifenthrin-treated zones at the rate 2.087 kg of formulated product/92.88 m2 and investigated the survival of individual ants successfully crossing the respective zones. Granular bifenthrin was nonrepellent to fire ant foragers in the laboratory. The 2.0- and 3.0-m treatment zones provided 100% protection for 7 wk after treatment and provided a reduction in the number of ants breaching the treated zone compared with the control for the remaining 9 wk of the study. This level of control may be tolerable for homeowners and is, therefore, considered an effective treatment for 15 wk after treatment. Hospitals, nursing homes, and electrical boxes would have to be treated on a monthly or bimonthly to remain ant free. PMID- 12650352 TI - Nutritional ecology of the Formosan subterranean termite (Isoptera: Rhinotermitidae): growth and survival of incipient colonies feeding on preferred wood species. AB - The wood of 11 plant species was evaluated as a food source significantly impacting the growth and survival of incipient colonies of the Formosan subterranean termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae). Colonies of C. formosanus feeding on pecan, Carya illinoensis (Wangenh.), and red gum, Liquidambar styraciflua L., produced significantly more progeny than colonies feeding on other wood species tested. Progeny of colonies feeding on pecan and American ash, Fraxinus americana L., had significantly greater survival than progeny of colonies feeding on other wood species. Colonies feeding on a nutritionally supplemented cellulose based matrix showed similar fitness characteristics as colonies feeding on the best wood treatments. These results indicate that differences observed in colony fitness can be partially explained by nutritional value of the food treatment, raising the possibility that wood from different tree species have different nutritional values to the Formosan subterranean termites. Colonies feeding on loblolly pine, Pinus taeda L., and ponderosa pine, Pinus ponderosa Laws., had significantly lower survival and produced significantly fewer workers and soldiers than colonies feeding on other wood species. Colony survival from 90 to 180 d of age and from 90 to 360 d of age was significantly correlated with the number of workers present at 90 d of colony age, indicating that colony survival depends on the presence of workers. Wood consumption in a multiple-choice study was significantly correlated with colony fitness value. This suggests that feeding preference of C. formosanus is at least partially influenced by the nutritional value of the food source. PMID- 12650353 TI - Effect of imidacloprid tree treatments on the occurrence of Formosan subterranean termites, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), in independent monitors. AB - Periodic sampling of 87 independent monitors, initially active with the Formosan subterranean termite, Coptotermes formosanus Shiraki, was conducted. Monitors, located in eight sectors adjacent to seven buildings, were various distances (1 46 m) from 57 trees treated with 0.1% imidacloprid foam. Termites collected from six of the eight sectors showed latent mortality attributed to imidacloprid intoxication at all monitor-tree distances. Approximately 6 mo after treatment, termite populations had recovered in these sectors. Another sector showed termite population suppression for approximately 15 mo, followed by recovery. Imidacloprid tree treatments did not control C. formosanus populations in independent monitors adjacent to the treatments. PMID- 12650354 TI - Controlling the mealworm Alphitobius diaperinus (Coleoptera: Tenebrionidae) in broiler and turkey houses: field trials with a combined insecticide treatment: insect growth regulator and pyrethroid. AB - Trials were conducted during one year under field conditions to control the lesser mealworm, Alphitobius diaperinus (Panzer), in broiler and turkey houses. The tested combined treatment included an adulticidal compound (pyrethroid: cyfluthrin) and a larvicidal compound (insect growth regulator [IGR]: triflumuron). The combined insecticide treatment greatly reduced the adult and larval stocks throughout the different broiler growing periods, and control of A. diaperinus populations was achieved by the end of the second treatment. Control of the insect population in a turkey house was not similar. A reestablishment of the insect population was observed during the second turkey growing period in summer. Building characteristics and management practices of the breeding system (duration of the breeding period, management of the litter) interact with the combined insecticide treatment and lead to a different efficiency. PMID- 12650355 TI - Inheritance of methyl-parathion resistance in Nebraska western corn rootworm populations (Coleoptera: Chrysomelidae). AB - Field populations of western corn rootworm, Diabrotica virgifera virgifera LeConte, were collected from three different sites (York Co., Phelps Co., and Saunders Co.) in Nebraska during 1996. Adult bioassays of these three populations were conducted with different concentrations of methyl-parathion and at a diagnostic concentration (1.0 microg/ml) to determine resistance levels among these populations. Self and reciprocal crosses were made between the two resistant and one susceptible laboratory-reared populations. Dose-responses and dominance ratios calculated for the four reciprocal crosses indicated that resistance was incompletely dominant in both strains, although in one of the strains there was an indication of sex linkage. However, evaluation of native polyacrylamide gels stained for nonspecific esterases and nonspecific esterase activity of parents and F1 progeny of the crosses suggested that esterase inheritance was completely dominant and autosomal. The results of this study were inconclusive with regard to the precise nature of inheritance, because the bioassays and esterase assays could not discriminate between heterozygotes and homozygotes. However, they do provide insight into the potential for developing simple diagnostic assays to assess resistance frequencies. Based on the inheritance studies described in this investigation, we can begin to generate information on specific genetic factors that dictate the evolutionary divergence of discrete resistant populations and facilitate modeling efforts designed to approximate the movement of genes for resistance among populations. PMID- 12650356 TI - Estimated frequency of nonrecessive Bt resistance genes in bollworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) in eastern North Carolina. AB - In summer 2000, adult female bollworm moths, Helicoverpa zea (Boddie), were collected from light-traps at four locations near the Tidewater Research Station, Plymouth, NC. Female moths were allowed to lay eggs, and at hatch, 72 larvae from each female were screened for growth rate on normal artificial diet and on diets containing 5.0 microg of either Cry1Ac or Cry2Aa Bt toxin per milliliter of diet. The growth rate bioassays were performed to isolate nonrecessive Bt resistance genes present in field populations of bollworm. We found one individual out of 583 screened that appeared to carry a major gene for resistance to Cry1Ac. Assuming four alleles per individual, the gene frequency is 1/2332 or 0.0003. Other females appeared to have minor genes for Cry1Ac resistance or major genes with lower levels of dominance. We also found one individual out of 646 screened that appeared to carry a major gene for resistance to Cry2Aa. The gene frequency for Cry2Aa resistance was estimated at 1/2584 or 0.00039. Again, other females seemed to carry additional minor resistance genes. Along with other results that indicate partially dominant inheritance of Cry1Ac resistance in bollworm, these allele frequency estimates are important for determining the rate of resistance evolution in H. zea to specific Bt toxins. PMID- 12650357 TI - Resistance of kale populations to lepidopterous pests in northwestern Spain. AB - Kale (Brassica oleracea L. acephala) is common in northwestern Spain where it is severely damaged by different insect pests. Damage could be reduced by using resistant varieties. The objectives of this work were to evaluate the resistance of kale populations to leaf damage by lepidopterous pests, to determine which traits are the best indicators of resistance, and finally to study the relationship between the glossy phenotype and resistance. Fifteen kale populations, sowed early and late, were evaluated at two locations in northwestern Spain. Significant differences among genotypes were found for all damage traits. Damage was not related to planting dates. Highest levels of damage were observed from July to November. Some populations with different performance under natural infestation in 1999 were again evaluated in 2000 under artificial infestation with Mamestra brassicae (L.) eggs. Two accessions, MBC-BRS0142 and MBG-BRS0170, showed resistance to attack by lepidopterous pests. Correlation coefficients among damage traits show that general appearance rating may be an useful indicator of resistance. Phenotype of kale with glossy leaves seems to be related to resistance although further research is needed. PMID- 12650358 TI - Resistance to Fiji disease in sugar cane: role of cultivar preference by planthopper vector Perkinsiella saccharicida (Homoptera: Delphacidae). AB - Fiji disease (FD) of sugar cane caused by Fiji disease virus (FDV) is transmitted by the planthopper Perkinsiella saccharicida Kirkaldy (Hemiptera Delphacidae). FD is effectively managed by using resistant cultivars, but whether the resistance is for the vector or for the virus is unknown. This knowledge would help develop a rapid and reliable glasshouse-based screening method for disease resistance. Sugar cane cultivars resistant, intermediate, and susceptible to FD were screened in a glasshouse, and the relationship between vector preferences and FD incidence was studied. Cultivar preference by nymphs increased with an increase in cultivar susceptibility to FD, but the relationship between adult preference and FD resistance was not significant. There was a positive correlation between the vector population and FD incidence, and the latent period for symptom expression declined with the increase in the vector populations. FD incidence in the glasshouse trial reflected the field-resistance status of sugar cane cultivars with known FD-resistance scores. The results suggest that resistance to FD in sugar cane is mediated by cultivar preference of the planthopper vector. PMID- 12650359 TI - Spatial processes in the evolution of resistance in Helicoverpa zea (Lepidoptera: Noctuidae) to Bt transgenic corn and cotton in a mixed agroecosystem: a biology rich stochastic simulation model. AB - A simulation model is developed to examine the role of spatial processes in the evolution of resistance in Helicoverpa zea populations to Bt corn and Bt cotton. The model is developed from the stochastic spatially explicit Heliothis virescens model described by Peck et al. (1999), to accommodate a spatial mix of two host crops (corn and cotton), and to reflect the agronomic practices, as well as the spatial and temporal population dynamics of H. zea, in eastern North Carolina. The model suggests that selection for resistance is more intense in Bt cotton fields than in Bt corn fields. It further suggests that local gene frequencies are highly dependent on local deployment levels of Bt crops despite the high mobility of the adult insects. Region-wide average gene frequencies depend on the region-wide level of Bt deployment, so incomplete technology adoption slows the rate of resistance evolution. However, on a local scale, H. zea populations in clusters of fields in which Bt use is high undergo far more rapid evolution than populations in neighboring clusters of fields in which Bt use is low. The model suggests that farm-level refuge requirements are important for managing the risk of resistance. The model can be used as an aid in designing plans for monitoring for resistance by suggesting the appropriate distribution of monitoring locations, which should focus on areas of highest Bt crop deployment. The findings need to be placed in the context of the input parameters, many of which are uncertain or highly variable in nature, and therefore, a thorough sensitivity analysis is warranted. PMID- 12650360 TI - Sensitivity analysis of a spatially-explicit stochastic simulation model of the evolution of resistance in Helicoverpa zea (Lepidoptera: Noctuidae) to Bt transgenic corn and cotton. AB - The sensitivities of a model simulating the evolution of resistance in Helicoverpa zea to Bt toxins in transgenic crops were investigated by examining effects of each of the model parameters on the frequency of resistance alleles after 8 yr. The functional dominance of resistance alleles and the initial frequency of those alleles had a major impact on resistance evolution. The survival of susceptible insects on the transgenic crops and the population dynamics of the insect, driven by winter survival and reproductive rates, were also important. In addition, agricultural practices including the proportion of the acreage planted to corn, and the larval threshold for spraying cotton fields affected the R-allele frequency. Many of these important parameters are inherently variable or cannot be measured with accuracy, so model output cannot be interpreted as being a forecast. However, this analysis is useful in focusing empirical research on those aspects of the insects' life system that have the largest effects on resistance development, and indicates ways in which to improve products and agricultural practices to increase the expected time to resistance. The model can thus be used as a scientific basis for devising a robust resistance management strategy for Bt crops. PMID- 12650361 TI - Photosynthetic responses of soybean to soybean aphid (Homoptera: Aphididae) injury. AB - The soybean aphid, Aphis glycines Matsumara, was discovered in the United States in the summer of 2000. Since that initial discovery, the aphid has spread across northern soybean production regions. In 2001, we examined the physiological responses of soybeans to low aphids densities (fewer than 50 aphids/leaf). In this study, we determined photosynthetic rates, leaf fluorescence responses, and photosynthetic responses to variable carbon dioxide and light levels. In addition, analyses for chlorophyll content and stable carbon isotope ratios were used to differentiate potential differences in stomatal versus mesophyll limitations to photosynthesis. We observed rate reductions of up to 50% on infested leaflets, including lealets with no apparent symptoms of aphid injury (such as chlorosis). Differences in fluorescence data indicated that photoelectron transport was not impaired. These results indicate that substantial physiological impact on soybean is possible even at low aphid densities. Also, the conventional view of aphid injury acting through reductions in chlorophyll content and light-harvesting reactions of photosynthesis is not supported by our findings in this system. PMID- 12650362 TI - Light activation of Russian wheat aphid-elicited physiological responses in susceptible wheat. AB - The impact of light and its role in Russian wheat aphid, Diuraphis noxia (Mordvilko), damage symptom formation, and photosynthetic capacity in 'Arapahoe' wheat (Triticum aestivum L.) were examined. After 72 h under continuous dark or continuous light regimes, the number of aphids (nymphs), leaf rolling and chlorosis ratings, fresh leaf weight, and chlorophyll contents were recorded. Photosynthetic rates, chlorophyll a, kinetics and chlorophyll extractions also were determined. Aphid infestation caused significant reductions in plant height, fresh weight, gas exchange, and chlorophyll fluorescence only under continuous light. Under the 72 h continuous dark regime, aphid infestation did not cause either damage symptom formation or reduction in plant growth or metabolism (photosynthesis). Furthermore, significantly more D. noxia nymphs were produced under continuous light condition than continuous dark. Our results demonstrate that the development of D. noxia feeding damage symptoms (i.e., leaf rolling and chlorotic streaks) on susceptible wheat seedlings is a light-activated process, even though the elicitor of the plant damage symptoms is aphid feeding. PMID- 12650363 TI - Expression and suppression of resistance to greenbug (Homoptera: Aphididae) in synthetic hexaploid wheats derived from Triticum dicoccum x Aegilops tauschii crosses. AB - Fifty-eight synthetic hexaploid wheats, developed by crossing Triticum dicoccum Schrank. and Aegilops tauschii (Coss.) Schmal., were evaluated at the seedling stage, together with their parents, for resistance to greenbug (Schizaphis graminum Rondani) under greenhouse conditions. Seedlings of different synthetic hexaploids showed large phenotypic differences for resistance. All the T. dicoccum parents were susceptible, while high levels of resistance were observed in some of the Ae. tauschii parents. Of the synthetic hexaploids derived from resistant Ae. tauschii parents, a high proportion (76%) showed levels of resistance to the greenbug biotype used that were comparable to those of the resistant parent. While there were clear indications of the presence of suppressor genes for greenbug resistance in the A and/or B genomes of T. dicoccum in some synthetics, positive epistatic interaction was also found in synthetic hexaploids with higher levels of resistance than that of either parent. Resistance from different Ae. tauschii accessions was expressed differently when crossed with the same T. dicoccum, indicating diversity among the resistance genes present in the test synthetic hexaploid wheats. Based on resistance reactions, the genes conferring greenbug resistance in these synthetic hexaploids are probably different from resistance genes previously transferred to wheat from Ae. tauschii. PMID- 12650364 TI - Characterization of resistance to three bruchid species (Callosobruchus spp., Coleoptera, Bruchidae) in cultivated rice bean (Vigna umbellata). AB - Resistance of wild and cultivated rice bean (Vigna umbellata [Thunberg] Ohwi and Ohashi) to three bruchid species, Callosobruchus chinensis L., Callosobruchus maculatus F., and Callosobruchus analis F., was evaluated. All but three accessions of cultivated, and all wild rice bean accessions tested, exhibited complete resistance to all three bruchid species. Rice bean seeds with seed coat removed also showed complete resistance to the three bruchid species. Results indicate that physical attributes and/or chemical(s) present in the seed coat of rice bean are not the main factors responsible for resistance. Feeding tests were performed by using artificial beans prepared with varying proportions of rice bean (resistant) and azuki bean (susceptible) flour. Number of bruchid adults that emerged decreased, and larval developmental period (days) was extended, when artificial beans with an increasing proportion of rice bean flour were used. These tests revealed that a chemical compound(s) contained in the cotyledon of rice bean has an inhibitory effect on the growth of these bruchid species. The results also indicate that the chemical(s) in rice bean cotyledon is most effective against C. maculatus. PMID- 12650365 TI - Categories of resistance at different growth stages in halt, a winter wheat resistant to the Russian wheat aphid (Homoptera: Aphididae). AB - This study was designed to categorize the resistance to the Russian wheat aphid, Diuraphis noxia (Mordvilko), resistant hard red winter wheat, Halt, as compared with susceptible wheat, TAM 107, at four different growth stages. Antixenosis was expressed in Halt at growth stage Zadoks 30. Antibiosis in Halt affected fecundity, number of aphids produced per reproductive day, maximum number of nymphs produced in one day, and intrinsic rate of increase. Fecundity was lower on Halt than TAM 107, and more nymphs were produced on both varieties at growth stage 20 than 10 and 40. Fewer nymphs were produced per reproductive day and on maximum production days by aphids reared on Halt than by those reared on TAM 107. The intrinsic rate of increase of Russian wheat aphids reared on Halt was lower than aphids reared on TAM 107. Differences in plant height and plant dry weight did not occur. Chlorosis ratings showed greater damage at the earlier stages in Halt and TAM 107 and significantly more damage in TAM 107 than Halt at growth stages 10, 20, and 30. Leaf rolling occurred on infested plants of TAM 107 at growth stages 10, 20, and 30, but not growth stage 40. Halt plants did not exhibit leaf rolling. The presence of a significant level of tolerance could make Halt compatible with other integrated pest management programs. However, care should be taken with cultivars containing evidence of antixenosis or antibiosis that could cause selective pressure on the Russian wheat aphid, potentially causing biotypes to be produced. PMID- 12650366 TI - Postharvest resistance in hard spring and winter wheat varieties of the northern Great Plains to the lesser grain borer (Coleoptera: Bostrichidae). AB - Wheat (Triticum aestivum L.) varietal resistance to the lesser grain borer, Rhyzopertha dominica (F), was evaluated in hard spring and winter wheat produced 1997, 1998 (Bozeman, Montana). We tested the hypothesis that six Montana-grown spring wheat varieties, 'Ernest', 'Scholar', 'Hi-Line', 'McNeal', 'Newana', and 'Amidon', were equally and strongly resistant to R. dominica at low moisture contents (9-10%). Mortality/Feeding damage occurred in all varieties. In most assays, Ernest had significantly greater feeding damage from R. dominica than other varieties, usually not significantly different from the susceptible control. Mean adult mortality was significantly greater in McNeal (93%) and Hi Line (92%) than in Ernest (34%) and Montana-grown, soft white spring wheat (Penawawa), the susceptible control (36%). In 9 wk, twice as many adult progeny were produced on Ernest than on McNeal, Hi-Line, or Scholar. We also compared three Montana-grown winter wheat varieties for resistance to R. dominica attack at low moisture contents (9-10%). Significantly more mortality after 6 wk was associated with all winter wheat varieties than the susceptible control. In 'Nuwest', 'Rocky', and 'Vanguard', significantly fewer progeny were produced than in the susceptible control; these varieties appeared more resistant. 'Tiber' and 'Neeley', in contrast, appeared more susceptible than other winter wheat varieties evaluated. Susceptibility decreased significantly with a 1.2% decrease in moisture content. Percentage of total protein was positively correlated with percentage of sound kernels and negatively with total progeny (r2 = -0.69). Kernel hardness was positively correlated with percentage of sound kernels, but negatively correlated with total progeny (r2 = -0.87) and dry weight loss. PMID- 12650367 TI - Comparison of the efficacy of different food attractants and their concentration for melon fly (Diptera: Tephritidae). AB - The relative attractiveness of six commercially available protein hydrolysates and the influence of their concentration were evaluated in field cages by a release-capture method of lab-reared melon fly adults, Bactrocera cucurbitae (Coquillett). Buminal, Corn Steepwater, Hym-Lure, Pinnacle, Nulure, and SolBait were tested for both sexes of the melon fly. The tested products exhibited clear differences in attractiveness. SolBait was the most effective protein hydrolysate. Pinnacle and Corn Steepwater also gave promising results. A general tendency for an increase in effectiveness with increasing concentration within the range 0.5 to 10% was shown. This study will allow pest control practitioners to choose more effective attractants for use in bait sprays to control the melon fly thus reducing the intensive use of insecticides currently practiced in Reunion island and enabling the development of Integrated pest management (IPM) methods for cucurbit crops. PMID- 12650368 TI - Potential of yellow sticky traps for lady beetle survey in cotton. AB - A 2-yr study was conducted to investigate the potential of using yellow sticky traps to survey lady beetles in cotton and to quantify seasonal activity patterns. The performance of sticky traps was compared with that of a 2-cycle vacuum sampler. The most common lady beetle species captured by sticky traps and vacuum sampler in cotton were Hippodamia convergens Guerin-Meneville and Scymnus loewii Mulsant. Sticky traps captured significantly more of both species of lady beetles, had greater capture efficiency, and more effectively detected lady beetles compared with the vacuum sampler. These data indicate that the sticky trap can be a valuable tool in monitoring lady beetle populations in cotton. In the second part of this study, a year-round survey of lady beetle populations in the periphery of a cotton farm using sticky traps showed that lady beetles remained active throughout the year in the Texas Rolling Plains, but the activity was influenced by winter severity. Over a 2-yr period, H. convergens, S. loewii, Coccinella septempunctata (L.), and Olla v-nigrum (Mulsant) comprised 89.6, 8.2, 1.9, and 0.3% of the specimens, respectively. Sticky trap captures were affected by year, trap height, and cropping season. Traps placed at 0.75 m above ground captured significantly more (80%) lady beetles than traps placed at 1.50 m (20%) above ground; traps at 0.75 m above ground also detected the rarer species while the traps at 1.50 m above ground detected only the abundant species. Trap captures were higher during the noncotton season (November to April) compared with the cotton season (May to October). A significant positive correlation between cotton aphid abundance during the growing season and H. convergens abundance during the following noncotton season was also detected, indicating a significant movement of H. convergens from cotton to the periphery of the farm to seek refuge after cotton termination. PMID- 12650369 TI - Efficacy of Silwet L-77 against several arthropod pests of table grape. AB - Silwet L-77, an organosilicone surfactant, was applied to several arthropod pests of California table grapes. Eggs of grape mealybug, Pseudococcus maritimus (Ehrhorn), and omnivorous leafroller, Platynota stultana Walsingham, were tolerant to 0.1, 0.25, and 0.5% treatment solutions; however, eggs of Pacific spider mite, Tetranychus pacificus McGregor, were highly susceptible with mortality >99.4% (0.1% Silwet L-77). Mortality of immature and adult stages of cotton aphid (Aphis gossypii Glover), Western flower thrips (Frankliniella occidentalis Pergande), and Pacific spider mite (Tetranychus pacificus McGregor) was > or = 93.8, > or = 98.5, and > or = 99.4% for 0.1, 0.25, and 0.5% Silwet L 77, respectively. Grape mealybug crawlers had 100% mortality when treated with 0.5 and 1.0% Silwet L-77 solutions; however, mortality was only 6.7% when 0.1% Silwet L-77 was applied. 'Thompson Seedless' table grapes were not damaged when treated with up to 1% Silwet L-77; however, grapes treated with the 0.5 and 1.0% solutions appeared wet after removal from cold storage because of the effect of the surfactant spreading the water condensation. Grapes dried with the normal bloom on the berries when they reached room temperature. PMID- 12650370 TI - The end of an era: what became of the "managed care revolution" in 2001? AB - OBJECTIVE: To describe how the organization and dynamics of health systems changed between 1999 and 2001, in the context of expectations from the mid-1990s when managed care was in ascendance, and assess the implications for consumers and policymakers. DATA SOURCES/STUDY SETTING: Data are from the Community Tracking Study site visits to 12 communities that were randomly selected to be nationally representative of metropolitan areas with 200,000 people or more. The Community Tracking Study is an ongoing effort that began in 1996 and is fielded every two years. STUDY DESIGN: Semistructured interviews were conducted with 50 90 stakeholders and observers of the local health care market in each of the 12 communities every two years. Respondents include leaders of local hospitals, health plans, and physician organizations and representatives of major employers, state and local governments, and consumer groups. First round interviews were conducted in 1996-1997 and subsequent rounds of interviews were conducted in 1998 1999 and 2000-2001. A total of 1,690 interviews were conducted between 1996 and 2001. DATA ANALYSIS METHODS: Interview information was stored and coded in qualitative data analysis software. Data were analyzed to identify patterns and themes within and across study sites and conclusions were verified by triangulating responses from different respondent types, examining outliers, searching for disconfirming evidence, and testing rival explanations. PRINCIPAL FINDINGS: Since the mid-1990s, managed care has developed differently than expected in local health care markets nationally. Three key developments shaped health care markets between 1999 and 2001: (1) unprecedented, sustained economic growth that resulted in extremely tight labor markets and made employers highly responsive to employee demands for even fewer restrictions on access to care; (2) health plans increasingly moved away from core strategies in the "managed care toolbox"; and (3) providers gained leverage relative to managed care plans and reverted to more traditional strategies of competing for patients based on services and amenities. CONCLUSIONS: Changes in local health care markets have contributed to rising costs and created new access problems for consumers. Moreover, the trajectory of change promises to make the goals of cost-control and quality improvement more difficult to achieve in the future. PMID- 12650371 TI - Managing costs, managing benefits: employer decisions in local health care markets. AB - OBJECTIVES: To better understand employer health benefit decision making, how employer health benefits strategies evolve over time, and the impact of employer decisions on local health care systems. DATA SOURCES/STUDY SETTING: Data were collected as part of the Community Tracking Study (CTS), a longitudinal analysis of health system change in 12 randomly selected communities. STUDY DESIGN: This is an observational study with data collection over a six-year period. DATA COLLECTION/EXTRACTION METHODS: The study used semistructured interviews with local respondents, combined with monitoring of local media, to track changes in health care systems over time and their impact on community residents. Interviewing began in 1996 and was carried out at two-year intervals, with a total of approximately 2,200 interviews. The interviews provided a variety of perspectives on employer decision making concerning health benefits; these perspectives were triangulated to reach conclusions. PRINCIPAL FINDINGS: The tight labor market during the study period was the dominant consideration in employer decision making regarding health benefits. Employers, in managing employee compensation, made independent decisions in pursuit of individual goals, but these decisions were shaped by similar labor market conditions. As a result, within and across our study sites, employer decisions in aggregate had an important impact on local health care systems, although employers' more highly visible public efforts to bring about health system change often met with disappointing results. CONCLUSIONS: General economic conditions in the 1990s had an important impact on the configuration of local health systems through their effect on employer decision making regarding health benefits offered to employees, and the responses of health plans and providers to those decisions. PMID- 12650372 TI - An empty toolbox? Changes in health plans' approaches for managing costs and care. AB - OBJECTIVE: To examine how health plans have changed their approaches for managing costs and utilization in the wake of the recent backlash against managed care. DATA SOURCES/STUDY SETTING: Semistructured interviews with health plan executives, employers, providers, and other health care decision makers in 12 metropolitan areas that were randomly selected to be nationally representative of communities with more than 200,000 residents. Longitudinal data were collected as part of the Community Tracking Study during three rounds of site visits in 1996 1997, 1998-1999, and 2000-2001. STUDY DESIGN: Interviews probed about changes in the design and operation of health insurance products--including provider contracting and network development, benefit packages, and utilization management processes--and about the rationale and perceived impact of these changes. DATA COLLECTION/EXTRACTION METHODS: Data from more than 850 interviews were coded, extracted, and analyzed using computerized text analysis software. PRINCIPAL FINDINGS: Health plans have begun to scale back or abandon their use of selected managed care tools in most communities, with selective contracting and risk contracting practices fading most rapidly and completely. In turn, plans increasingly have sought cost savings by shifting costs to consumers. Some plans have begun to experiment with new provider networks, payment systems, and referral practices designed to lower costs and improve service delivery. CONCLUSIONS: These changes promise to lighten administrative and financial burdens for physicians and hospitals, but they also threaten to increase consumers' financial burdens. PMID- 12650373 TI - Medicare contracting risk/Medicare risk contracting: a life-cycle view from twelve markets. AB - OBJECTIVE: To examine the evolution of the Medicare HMO program from 1996 to 2001 in 12 nationally representative urban markets by exploring how the separate and confluent influences of government policy initiatives and health plans' strategic aims and operational experience affected the availability of HMOs to Medicare beneficiaries. DATA SOURCE: Qualitative data gathered from 12 nationally representative urban communities with more than 200,000 residents each, in tandem with quantitative information from the Centers for Medicare and Medicaid Services and other sources. STUDY DESIGN: Detailed interview protocols, developed as part of the multiyear, multimethod Community Tracking Study of the Center for Studying Health System Change, were used to conduct three rounds of interviews (1996, 1998, and 2000-2001) with health plans and providers in 12 nationally representative urban communities. A special focus during the third round of interviews was on gathering information related to Medicare HMOs' experience in the previous four years. This information was used to build on previous research to develop a longitudinal perspective on health plans' experience in Medicare's HMO program. PRINCIPAL FINDINGS: From 1996 to 2001, the activities and expectations of health plans in local markets underwent a rapid and dramatic transition from enthusiasm for the Medicare HMO product, to abrupt reconsideration of interest corresponding to changes in the Balanced Budget Act of 1997, on to significant retrenchment and disillusionment. Policy developments were important in their own right, but they also interacted with shifts in the strategic aims and operational experiences of health plans that reflect responses to insurance underwriting cycle pressures and pushback from providers. CONCLUSION: The Medicare HMO program went through a substantial reversal of fortune during the study period, raising doubts about whether its downward course can be altered. Market-level analysis reveals that virtually all momentum for the program has been lost and that enrollment is shrinking back to the levels and locations found in the mid-1990s. PMID- 12650374 TI - Hospitals' negotiating leverage with health plans: how and why has it changed? AB - OBJECTIVE: To describe how hospitals' negotiating leverage with managed care plans changed from 1996 to 2001 and to identify factors that explain any changes. DATA SOURCES: Primary semistructured interviews, and secondary qualitative (e.g., newspaper articles) and quantitative (i.e., InterStudy, American Hospital Association) data. STUDY DESIGN: The Community Tracking Study site visits to a nationally representative sample of 12 communities with more than 200,000 people. These 12 markets have been studied since 1996 using a variety of primary and secondary data sources. DATA COLLECTION METHODS: Semistructured interviews were conducted with a purposive sample of individuals from hospitals, health plans, and knowledgeable market observers. Secondary quantitative data on the 12 markets was also obtained. PRINCIPAL FINDINGS: Our findings suggest that many hospitals' negotiating leverage significantly increased after years of decline. Today, many hospitals are viewed as having the greatest leverage in local markets. Changes in three areas--the policy and purchasing context, managed care plan market, and hospital market--appear to explain why hospitals' leverage increased, particularly over the last two years (2000-2001). CONCLUSIONS: Hospitals' increased negotiating leverage contributed to higher payment rates, which in turn are likely to increase managed care plan premiums. This trend raises challenging issues for policymakers, purchasers, plans, and consumers. PMID- 12650375 TI - Changes in hospital competitive strategy: a new medical arms race? AB - OBJECTIVE: To describe changes in hospitals' competitive strategies, specifically the relative emphasis placed on strategies for competing along price and nonprice (i.e., service, amenities, perceived quality) dimensions, and the reasons for any observed shifts. METHODS: This study uses data gathered through the Community Tracking Study site visits, a longitudinal study of a nationally representative sample of 12 U.S. communities. Research teams visited each of these communities every two years since 1996 and conducted between 50 to 90 semistructured interviews. Additional information on hospital competition and strategy was gathered from secondary data. PRINCIPAL FINDINGS: We found that hospitals' strategic emphasis changed significantly between 1996-1997 and 2000-2001. In the mid-1990s, hospitals primarily competed on price through "wholesale" strategies (i.e., providing services attractive to managed care plans). By 2000-2001, nonprice competition was becoming increasingly important and hospitals were reviving "retail" strategies (i.e., providing services attractive to individual physicians and the patients they serve). Three major factors explain this shift in hospital strategy: less than anticipated selective contracting and capitated payment; the freeing up of hospital resources previously devoted to horizontal and vertical integration strategies; and, the emergence and growth of new competitors. CONCLUSION: Renewed emphasis on nonprice competition and retail strategies, and the service mimicking and one-upmanship that result, suggest that a new medical arms race is emerging. However, there are important differences between the medical arms race today and the one that occurred in the 1970s and early 1980s: the hospital market is more concentrated and price competition remains relatively important. The development of a new medical arms race has significant research and policy implications. PMID- 12650376 TI - Something old, something new: recent developments in hospital-physician relationships. AB - OBJECTIVE: To describe recent developments in hospital-physician relationships in 12 metropolitan areas. METHODS: We analyze qualitative data from a third round of biannual site visit interviews conducted in 12 randomly selected metropolitan areas from 1996 to 2001. The study interviewed 895 respondents during the third round of site visits, conducted in 2000 and 2001. PRINCIPAL FINDINGS: As HMO enrollment and capitation contracting has failed to grow in local markets, hospital executives have returned to a strategic focus on improving relationships with specialists in pursuit of fee-for-service revenue. Yet, 65 percent of hospitals interviewed in 2000 and 2001 continued to own primary care physician practices, with ownership more prevalent in highly concentrated hospital markets. A majority (55 percent) of hospitals have decreased the size of these practices in the past two years. CONCLUSIONS: Interest in forming integrated delivery systems has waned. The potential for quality improvement through these organizations systems--by emphasizing primary care and coordinating hospital and physician services--has not been realized. The new emphasis on hospital specialist partnerships may improve the financial status of hospitals and participating specialists in local markets, and may improve quality of care in selected service areas, but it may also increase health care costs incurred by employers and consumers. PMID- 12650377 TI - The resilience of the health care safety net, 1996-2001. AB - OBJECTIVE: To determine how the capacity and viability of local health care safety nets changed over the last six years and to draw lessons from these changes. DATA SOURCE: The first three rounds (May 1996 to March 2001) of Community Tracking Study site visits to 12 communities. STUDY DESIGN: Researchers visited the study communities every two years to interview leaders of local health care systems about changes in the organization, delivery, and financing of health care and the impact of these changes on people. For this analysis, we collected data on safety net capacity and viability through interviews with public and not-for-profit hospitals, community health centers, health departments, government officials, consumer advocates, academics, and others. We asked about the effects of market and policy changes on the safety net and how the safety net responded, as well as the impact of these changes on care for the low-income uninsured. PRINCIPAL FINDINGS: The safety net in three-quarters of the communities was stable or improved by the end of the study period, leading to improved access to primary and preventive care for the low-income uninsured. Policy responses to pressures such as the Balanced Budget Act and Medicaid managed care, along with effective safety net strategies and supportive conditions, helped reinforce the safety net. However, the safety net in three sites deteriorated and access to specialty services remained inadequate across the 12 sites. CONCLUSIONS: Despite pessimistic predictions and some notable exceptions, the health care safety net grew stronger over the past six years. Given considerable community variation, however, this analysis indicates that policymakers can apply a number of lessons from strong and improving safety nets to strengthen those that are weaker, particularly as the current economy poses new challenges. PMID- 12650378 TI - Managed care, access to specialists, and outcomes among primary care patients with pain. AB - OBJECTIVE: To determine whether managed care controls were associated with reduced access to specialists and worse outcomes among primary care patients with pain. DATA SOURCES/STUDY SETTING: Patient, physician, and office manager questionnaires collected in the Seattle area in 1996-1997, plus data abstracted from patient records and health plans. STUDY DESIGN: A prospective cohort study of 2,275 adult patients with common pain problems recruited in the offices of 261 primary care physicians in Seattle. DATA COLLECTION: Patients completed a waiting room questionnaire and follow-up surveys at the end of the first and sixth months to measure access to specialists and outcomes. Intensity of managed care controls measured by plan managed care index and benefit/cost-sharing indexes, office managed care index, physician compensation, financial incentives, and use of clinical guidelines. PRINCIPAL FINDINGS: A financial withhold for referral was associated with a lower likelihood of referral to a physician specialist, a greater likelihood of seeing a specialist without referral, and a lower patient rating of care from the primary physician. Otherwise, patients in more managed offices and with greater out-of-network plan benefits had greater access to specialists. Patients with more versus less managed care had similar health outcomes, but patients in more managed offices had lower ratings of care provided by their primary physicians. CONCLUSIONS: Increased managed care controls were generally not associated with reduced access to specialists and worse health outcomes for primary care patients with pain, but patients in more managed offices had lower ratings of care provided by their primary physicians. PMID- 12650379 TI - Impact of cardiac service availability on case-selection for angiography and survival associated with angiography. AB - OBJECTIVE: To examine whether availability of cardiac services at the admitting hospital affects case-selection for angiography and one-year survival following angiography, within groups of patients who have similar clinical need for angiography according to published criteria. STUDY SETTING: Elderly Medicare beneficiaries (37,788) discharged with a diagnosis of acute myocardial infarction (AMI) from hospitals in seven U.S. states between February 1994 and July 1995. We focused on patients who were eligible to receive angiography 12 or more hours after symptom onset. DATA COLLECTION: Data were abstracted from patient's medical records, Medicare National Claims Standard Analytic Files, Health Care Financing Administration (HCFA) Provider of Service File and Health Insurance Master File. METHODS: Admitting hospitals were classified as offering no cardiac services, angiography only, or revascularization. Case-selection differences across these three types of hospitals were examined by comparing relative risk of receiving angiography for various patient and hospital characteristics. Relative differences in one-year survival rate, comparing patients who received angiography to those who did not, were estimated within each hospital type and clinical need category (necessary, appropriate, or uncertain) after matching on propensity to receive angiography. PRINCIPAL FINDINGS: Compared to patients for whom angiography was deemed necessary, the relative risk of receiving angiography among those for whom it was deemed of uncertain benefit was 0.58, 0.79, and 0.92 (p-value of homogeneity test < 0.001) at hospitals offering no cardiac services, angiography only, and revascularization, respectively. There was no significant difference in survival following angiography across hospital types, overall as well as within clinical need categories. CONCLUSIONS: Despite increased case selection at hospitals with on-site cardiac services, there was no evidence of increase in the survival rate associated with angiography use at these hospitals. PMID- 12650380 TI - Process of care and outcome after acute myocardial infarction for patients with mental illness in the VA health care system: are there disparities? AB - OBJECTIVE: To compare process of care and outcome after acute myocardial infarction, for patients with and without mental illness, cared for in the Veterans Health Administration (VA) health care system. DATA SOURCES/SETTING: Primary clinical data from 81 VA hospitals. STUDY DESIGN: This was a retrospective cohort study of 4,340 veterans discharged with clinically confirmed acute myocardial infarction. Of these, 859 (19.8 percent) met the definition of mental illness. Measures were age-adjusted in-hospital and 90-day cardiac procedure use; age-adjusted relative risks (RE) of use of thrombolytic therapy, beta-blockers, angiotensin converting enzyme (ACE) inhibitors, or aspirin at discharge; risk-adjusted 30-day and one-year mortality. RESULTS: Patients with mental illness were marginally less likely than those without mental illness to undergo in-hospital angiography (age-adjusted RR 0.90 [95 percent confidence interval: 0.83, 0.98]), but there was no significant difference in the age adjusted RR of coronary artery bypass graft surgery in the 90 days after admission (0.85 [0.69, 1.05]), or in the receipt of medications of known benefit. For example, ideal candidates with and without mental illness were equally likely to receive beta-blockers at the time of discharge (age-adjusted RR 0.92 [0.82, 1.02]). The risk-adjusted odds ratio (OR) for death in patients with mental illness versus those without mental illness within 30 days was 1.00 (0.75, 1.32), and for death within one year was 1.25 (1.00, 1.53). CONCLUSIONS: Veterans Health Administration patients with mental illness were marginally less likely than those without mental illness to receive diagnostic angiography, and no less likely to receive revascularization or medications of known benefit after acute myocardial infarction. Mortality at one year may have been higher, although this finding did not reach statistical significance. These findings are consistent with other studies showing reduced health care disparities in the VA for other vulnerable groups, and suggest that an integrated health care system with few financial barriers to health care access may attenuate some health care disparities. Further work should address how health care organizational features might narrow disparities in health care for vulnerable groups. PMID- 12650381 TI - Quality improvement implementation in the nursing home. AB - OBJECTIVE: To examine quality improvement (QI) implementation in nursing homes, its association with organizational culture, and its effects on pressure ulcer care. DATA SOURCES/STUDY SETTING: Primary data were collected from staff at 35 nursing homes maintained by the Department of Veterans Affairs (VA) on measures related to QI implementation and organizational culture. These data were combined with information obtained from abstractions of medical records and analyses of an existing database. STUDY DESIGN: A cross-sectional analysis of the association among the different measures was performed. DATA COLLECTION/EXTRACTION METHODS: Completed surveys containing information on QI implementation, organizational culture, employee satisfaction, and perceived adoption of guidelines were obtained from 1,065 nursing home staff. Adherence to best practices related to pressure ulcer prevention was abstracted from medical records. Risk-adjusted rates of pressure ulcer development were calculated from an administrative database. PRINCIPAL FINDINGS: Nursing homes differed significantly (p<.001) in their extent of QI implementation with scores on this 1 to 5 scale ranging from 2.98 to 4.08. Quality improvement implementation was greater in those nursing homes with an organizational culture that emphasizes innovation and teamwork. Employees of nursing homes with a greater degree of QI implementation were more satisfied with their jobs (a 1-point increase in QI score was associated with a 0.83 increase on the 5-point satisfaction scale, p<.001) and were more likely to report adoption of pressure ulcer clinical guidelines (a 1-point increase in QI score was associated with a 28 percent increase in number of staff reporting adoption, p<.001). No significant association was found, though, between QI implementation and either adherence to guideline recommendations as abstracted from records or the rate of pressure ulcer development. CONCLUSIONS: Quality improvement implementation is most likely to be successful in those VA nursing homes with an underlying culture that promotes innovation. While QI implementation may result in staff who are more satisfied with their jobs and who believe they are providing better care, associations with improved care are uncertain. PMID- 12650383 TI - Medicare home health utilization in context. PMID- 12650382 TI - Utilization of home health services before and after the Balanced Budget Act of 1997: what were the initial effects? AB - OBJECTIVE: To estimate the impact of the Balanced Budget Act of 1997 (BBA), which changed the way Medicare reimbursed for home health services, on a range of home health utilization measures, and to examine whether particular subgroups of beneficiaries were differentially impacted in the post-BBA period. DATA SOURCES: Secondary data from the Centers for Medicare and Medicaid Services (CMS) Standard Analytic Files for the 1 percent sample of Medicare beneficiaries for fiscal years 1997 and 1999, linked with information from CMS eligibility, provider, and cost report files as well as the Area Resources File. STUDY DESIGN: Logistic regression was used to estimate the effects of being in the post-BBA period on the incidence of home health service use and ordinary least squares (OLS) regression was used to estimate the effects of being in the post-BBA period on the amount and type of use by home health service users. Interaction terms we reincluded for all the independent variables to assess whether the effect was disproportionate among particular beneficiary subgroups. PRINCIPAL FINDINGS: Results show a 22 percent decrease in the percentage using home health services post-BBA and a 39 percent decrease in the number of visits per user. Stronger reductions, though not very large, were found in the incidence of use for beneficiaries aged 85 and older, those in states with high historical Medicare home health use, and those with Medicaid buy-in. More intensive reductions in the number of services were found for those aged 85 and older, in high historical Medicare use states, nonwhites, females, those using for-profit agencies, and those treated for certain diagnoses. Less intensive reductions were associated with hospital-based agencies. CONCLUSIONS: This research demonstrates that public program expenditures can be sharply curtailed with financial incentives. As reimbursement shifts to a prospective payment system legislated by the BBA, utilization should be closely monitored, especially for vulnerable subgroups. PMID- 12650384 TI - The effect of health plan characteristics on Medicare+ Choice enrollment. AB - OBJECTIVE: To provide national estimates of the effect of out-of-pocket premiums and benefits on Medicare beneficiaries' choice among managed care health plans. DATA SOURCES/STUDY SETTING: The data represent the population of all Medicare+ Choice (M+C) plans offered to Medicare beneficiaries in the United States in 1999. STUDY DESIGN: The dependent variable is the log of the ratio of the market share of the jth health plan to the lowest cost plan in the beneficiary's county of residence. The explanatory variables are measures of premiums and benefits in the jth health plan relative to the premiums and benefits in the lowest cost plan. DATA COLLECTION METHODS: The data are from the 1999 Medicare Compare database, and M+C enrollment data from the Centers for Medicare and Medicaid Services (CMS). PRINCIPAL FINDINGS: A $10 increase in an M+C plan's out-of-pocket premium, relative to its competitors, is associated with a decrease of four percentage points in the jth plan's market share (i.e., from 25 to 21 percent), holding the premiums of competing plans constant. CONCLUSIONS: Although our price elasticity estimates are low, the market share losses associated with small changes in a health plan's premium, relative to its competitors, may be sufficient to discipline premiums in a competitive market. Bidding behavior by plans in the Medicare Competitive Pricing Demonstration supports this conclusion. PMID- 12650385 TI - Is exposure to income inequality a public health concern? Lagged effects of income inequality on individual and population health. AB - OBJECTIVE: To examine the health consequences of exposure to income inequality. DATA SOURCES: Secondary analysis employing data from several publicly available sources. Measures of individual health status and other individual characteristics are obtained from the March Current Population Survey (CPS). State-level income inequality is measured by the Gini coefficient based on family income, as reported by the U.S. Census Bureau and Al-Samarrie and Miller (1967). State-level mortality rates are from the Vital Statistics of the United States, other state-level characteristics are from U.S. census data as reported in the Statistical Abstract of the United States. STUDY DESIGN: We examine the effects of state-level income inequality lagged from 5 to 29 years on individual health by estimating probit models of poor/fair health status for samples of adults aged 25-74 in the 1995 through 1999 March CPS. We control for several individual characteristics, including educational attainment and household income, as well as regional fixed effects. We use multivariate regression to estimate the effects of income inequality lagged 10 and 20 years on state-level mortality rates for 1990, 1980, 1970, and 1960. PRINCIPAL FINDINGS: Lagged income inequality is not significantly associated with individual health status after controlling for regional fixed effects. Lagged income inequality is not associated with all cause mortality, but associated with reduced mortality from cardiovascular disease and malignant neoplasms, after controlling for state fixed-effects. CONCLUSIONS: In contrast to previous studies that fail to control for regional variations in health outcomes, we find little support for the contention that exposure to income inequality is detrimental to either individual or population health. PMID- 12650386 TI - Income inequality as a public health concern: where do we stand? Commentary on "Is exposure to income inequality a public health concern?". PMID- 12650387 TI - Determinants of HMO formulary adoption decisions. AB - OBJECTIVE: To identify economic and organizational characteristics that affect the likelihood that health maintenance organizations (HMOs) include new drugs on their formularies. DATA SOURCES: We administered an original survey to directors of pharmacy at 75 HMOs, of which 41 returned usable responses. We obtained drug specific data from an industry trade journal. STUDY DESIGN: We performed multivariate logistic regression analysis, adjusting for fixed-drug effects and random-HMO effects. We used factor analysis to limit the number of predictors. DATA COLLECTION METHODS: We held initial focus groups to help with survey design. We administered the survey in two waves. We asked respondents to report on seven popular new drugs, and to describe a variety of HMO organizational characteristics. PRINCIPAL FINDINGS: Several HMO organizational characteristics, including nonprofit status, the incentives facing the director of the pharmacy, size and make-up of the pharmacy and therapeutics committee, and relationships with drugs makers, all affect formulary adoption. CONCLUSIONS: There are many organizational factors that may cause HMOs to make different formulary adoption decisions for certain prescription drugs. PMID- 12650388 TI - The effect of capitation on switching primary care physicians. AB - OBJECTIVE: To examine the relationship between patient case-mix, utilization, primary care physician (PCP) payment method, and the probability that patients switch their PCPs. DATA SOURCES/STUDY SETTING: Administrative enrollment and claims/encounter data for 1994-1995 from four physician organizations. STUDY DESIGN: We developed a conceptual model of patient switching behavior, which we used to guide the specification of multivariate logistic analyses focusing on interactions between patient case-mix, utilization, and PCP reimbursement methods. DATA COLLECTION/EXTRACTION METHODS: Claims data were aggregated to the encounter level; a switch was defined as a change in PCP since the previous encounter. The PCPs were reimbursed on either a capitated or fee-for-service (FFS) basis. PRINCIPAL FINDINGS: Patients with stable chronic conditions (Ambulatory Diagnostic Groups [ADG] 10) and capitated PCPs were 36 percent more likely to switch PCPs than similar patients with FFS PCPs, controlling for patient age and sex and physician fixed effects. When the number of previous encounters was included in the model this relationship was no longer significant. Instead high utilizers with capitated PCPs were significantly more likely to switch PCPs than were similar patients with FFS PCPs. CONCLUSIONS: A patient's demographics and utilization are associated with the probability that the patient will switch PCPs. Capitated PCP payment was associated with higher rates of switching among high utilizers of health care resources. These findings raise concerns about the continuity and quality of care experienced by vulnerable patients in an era of changing financial incentives. PMID- 12650389 TI - Racial and ethnic disparities in the purchase of nongroup health insurance: the roles of community and family-level factors. AB - OBJECTIVE: To evaluate the influence of community- and family-level factors on racial/ethnic disparities in the uptake of nongroup (individual) health insurance. DATA SOURCES: Responses to the 1996-1997 Community Tracking Study Household Survey plus community-level descriptors from several sources including census data, the Area Resource File, and community and migrant health center Medicare cost reports. STUDY DESIGN: Logistic regression was used to compare families in which at least one person had nongroup health insurance to families without nongroup insurance in which at least one person was uninsured. Sequential models were constructed examining family- and community-level factors. RESULTS: Twenty-three percent of families with otherwise-uninsured persons purchased nongroup insurance, ranging from 11% to 41% among the 60 communities sampled. Disadvantaged minority group members, especially Spanish-speaking Hispanics, had half or less the odds of whites of purchasing nongroup insurance. Education had a weaker association with purchasing nongroup insurance among minority group members than among whites. Community-level factors had minimal effect on disparities in uptake, although greater housing segregation was associated with lower uptake among blacks. CONCLUSIONS: Minority group members are much less likely to purchase nongroup insurance than whites. Family income and community factors do not explain this gap. Programs aimed at stimulating voluntary insurance purchase will continue to underenroll disadvantaged minorities if nonfinancial barriers to acquiring insurance coverage, including the interplay between race/ethnicity and education, are not better understood and addressed. PMID- 12650390 TI - Effectiveness and cost-effectiveness of four treatment modalities for substance disorders: a propensity score analysis. AB - OBJECTIVE: To assess the effectiveness and cost-effectiveness of four treatment modalities for substance abuse. DATA SOURCES: The study used data from the Services Research Outcomes Study (SROS), a survey of 3,047 clients in a random sample of 99 drug treatment facilities across the United States. Detailed sociodemographic, substance use, and clinical data were abstracted from treatment records. Substance abuse outcome and treatment history following discharge from index facilities were assessed using a comprehensive interview with 1,799 of these individuals five years after discharge. Treatment success was defined in two ways: as abstinence and as any reduction in substance use. STUDY DESIGN: Effectiveness and cost-effectiveness of four modalities were compared: inpatient, residential, outpatient detox/methadone, and outpatient drug-free. Clients were stratified based on propensity scores and analyses were conducted within these strata. Sensitivity analyses examined the impact of future substance abuse treatment on effectiveness and cost-effectiveness estimates. PRINCIPAL FINDINGS: Treatment of substance disorders appears to be cost-effective compared to other health interventions. The cost per successfully treated abstinent case in the least costly modality, the outpatient drug-free programs, was 6,300 dollars (95 percent confidence intervals: 5,200-7,900 dollars) in 1990 dollars. There were only minor differences between various modalities of treatment with regard to effectiveness. However, modalities varied considerably with regard to cost effectiveness. Outpatient drug-free programs were the most cost-effective. There was little evidence that relative effectiveness or cost-effectiveness of programs varied according to factors that were associated with selection into different programs. CONCLUSIONS: Substance disorders can be treated most cost-effectively in outpatient drug-free settings. Savings from transitioning to the most cost effective treatment modality may free resources that could be reinvested to improve access to substance abuse treatment for a larger number of individuals in need of such treatment. PMID- 12650391 TI - Factors that influence line managers' perceptions of hospital performance data. AB - OBJECTIVE: To design and test a model of the factors that influence frontline and midlevel managers' perceptions of usefulness of comparative reports of hospital performance. STUDY SETTING: A total of 344 frontline and midlevel managers with responsibility for stroke and medical cardiac patients in 89 acute care hospitals in the Canadian province of Ontario. STUDY DESIGN: Fifty-nine percent of managers responded to a mail survey regarding managers' familiarity with a comparative report of hospital performance, ratings of the report's data quality, relevance and complexity, improvement culture of the organization, and perceptions of usefulness of the report. EXTRACTION METHODS: Exploratory factor analysis was performed to assess the dimensionality of performance data characteristics and improvement culture. Antecedents of perceived usefulness and the role of improvement culture as a moderator were tested using hierarchical regression analyses. PRINCIPAL FINDINGS: Both data characteristics variables including data quality, relevance, and report complexity, as well as organizational factors including dissemination intensity and improvement culture, explain significant amounts of variance in perceptions of usefulness of comparative reports of hospital performance. The total R2 for the full hierarchical regression model = .691. Improvement culture moderates the relationship between data relevance and perceived usefulness. CONCLUSIONS: Organizations and those who fund and design performance reports need to recognize that both report characteristics and organizational context play an important role in determining line managers' response to and ability to use these types of data. PMID- 12650392 TI - Primary care service areas: a new tool for the evaluation of primary care services. AB - OBJECTIVE: To develop and characterize utilization-based service areas for the United States which reflect the travel of Medicare beneficiaries to primary care clinicians. DATA SOURCE/STUDY SETTING: The 1996-1997 Part B and 1996 Outpatient File primary care claims for fee-for-service Medicare beneficiaries aged 65 and older. The 1995 Medicaid claims from six states (1995) and commercial claims from Blue Cross Blue Shield of Michigan (1996). STUDY DESIGN: A patient origin study was conducted to assign 1999 U.S. zip codes to Primary Care Service Areas on the basis of the plurality of beneficiaries' preference for primary care clinicians. Adjustments were made to establish geographic contiguity and minimum population and service localization. Generality of areas to younger populations was tested with Medicaid and commercial claims. DATA COLLECTION/EXTRACTION METHODS: Part B primary care claims were selected on the basis of provider specialty, place of service, and CPT code. Selection of Outpatient File claims used provider number, type of facility/service, and revenue center codes. PRINCIPAL FINDINGS: The study delineated 6,102 Primary Care Service Areas with a median population of 17,276 (range 1,005-1,253,240). Overall, 63 percent of the Medicare beneficiaries sought the plurality of their primary care from within area clinicians. Service localization compared to Medicaid (six states) and commercial primary care utilization (Michigan) was comparable but not identical. CONCLUSIONS: Primary Care Service Areas are a new tool for the measurement of primary care resources, utilization, and associated outcomes. Policymakers at all jurisdictional levels as well as researchers will have a standardized system of geographical units through which to assess access to, supply, use, organization, and financing of primary care services. PMID- 12650394 TI - Failed anterior cervical foraminotomy. PMID- 12650393 TI - Catching up on health outcomes: the Texas Medication Algorithm Project. AB - OBJECTIVE: To develop a statistic measuring the impact of algorithm-driven disease management programs on outcomes for patients with chronic mental illness that allowed for treatment-as-usual controls to "catch up" to early gains of treated patients. DATA SOURCES/STUDY SETTING: Statistical power was estimated from simulated samples representing effect sizes that grew, remained constant, or declined following an initial improvement. Estimates were based on the Texas Medication Algorithm Project on adult patients (age > or = 18) with bipolar disorder (n = 267) who received care between 1998 and 2000 at 1 of 11 clinics across Texas. STUDY DESIGN: Study patients were assessed at baseline and three month follow-up for a minimum of one year. Program tracks were assigned by clinic. DATA COLLECTION/EXTRACTION METHODS: Hierarchical linear modeling was modified to account for declining-effects. Outcomes were based on 30-item Inventory for Depression Symptomatology-Clinician Version. PRINCIPAL FINDINGS: Declining-effect analyses had significantly greater power detecting program differences than traditional growth models in constant and declining-effects cases. Bipolar patients with severe depressive symptoms in an algorithm-driven, disease management program reported fewer symptoms after three months, with treatment-as-usual controls "catching up" within one year. CONCLUSIONS: In addition to psychometric properties, data collection design, and power, investigators should consider how outcomes unfold over time when selecting an appropriate statistic to evaluate service interventions. Declining-effect analyses may be applicable to a wide range of treatment and intervention trials. PMID- 12650395 TI - Failed anterior cervical foraminotomy. AB - OBJECT: Anterior cervical foraminotomy has been advocated as a procedure that preserves the motion segment while treating radiculopathy due to degenerative cervical disc disease. Because the medical literature contains no long-term follow up or randomized studies related to this procedure, the authors reviewed their results, specifically examining cases of failure to determine the efficacy of the approach. METHODS: The authors identified 23 patients in whom unilateral cervical radiculopathy due to degenerative cervical disc disease was refractory to conservative therapy and in whom anterior cervical foraminotomy was performed between 1998 and 2000. The procedure involves ipsilateral exposure, microsurgical removal of the uncovertebral joint to identify the nerve root, and partial removal of the lateral anulus and or disc fragments. Data in those patients who underwent reoperation(s) were reviewed specifically for the procedure type, interval between index procedure and reoperation, and whether multiple procedures were performed. Of the 23 patients, 30% required at least one additional procedure. A good or excellent outcome at last follow-up examination was achieved in only 12 patients. CONCLUSIONS: In the current study the authors found a reoperation rate that is considerably higher than that in most series of anterior cervical surgery for radiculopathy. The presumed benefit of anterior cervical foraminotomy is preservation of the disc interspace; however, in this study, a significant number of patients failed to experience a satisfying outcome. Currently the authors do not recommend anterior cervical foraminotomy as a stand alone procedure. PMID- 12650396 TI - Cervicothoracic radiculopathy treated using posterior cervical foraminotomy/discectomy. AB - OBJECT: The authors conducted a study to identify the effectiveness and morbidity rate associated with treating cervicothoracic disc disease (radiculopathy) via a posterior approach. METHODS: Nineteen patients underwent posterior cervicothoracic laminoforaminotomy during a 5.6-year period. Medical records, imaging studies, office charts, hospital records, and phone interview data were reviewed. Specific information analyzed included patient demographics, side of lesion, and conservative treatment, symptoms, and pre- and postoperative pain levels. Pain was rated using a visual analog scale and classified into a radicular and neck component. Data in 19 patients (seven women and 12 men) who underwent 20 procedures (one patient underwent separate bilateral foraminotomies) were analyzed. The mean patient age was 54.8 years (range 38-73 years), and the follow-up period ranged from 23 to 62 months. Symptom duration ranged from 1 to 14 months (mean 3.4 months) and consisted of weakness, numbness, and painful radiculopathies in 11, 16, and 20 cases, respectively. Motor weakness was identified in 11 of 19 patients (mean grade of 4.35), and postoperatively strength normalized in eight of 11 (mean grade of 4.79). The improvement in motor scores was significant (p = 0.007). Pain was the most common presenting symptom. Preoperative radiculopathies were rated between 0 and 10 (mean 7.45), and postoperatively scores were reduced to 0 to 3 (mean 0.2) which was significant (p < 0.0001). Preoperative neck pain was rated between 0 and 8 (mean 2.55), and on follow up ranged from 0 to 2 (mean 0.5), which was also significant (p = 0.001). CONCLUSIONS: Posterior cervicothoracic foraminotomy was a safe and effective procedure in the treatment of patients with laterally located disc herniations. PMID- 12650397 TI - Cervical laminoplasty: evaluation of bone bonding of a high porosity hydroxyapatite spacer. AB - OBJECT: The purpose of this study was to evaluate the usefulness of a high porosity hydroxyapatite (HA) spacer in cervical laminoplasty. Bone-spacer bonding rates, complications associated with the implant, and factors related to bone bonding were examined. METHODS: The authors evaluated 33 consecutive patients with cervical myelopathy who underwent high-porosity HA spacer-assisted laminoplasty and were followed for at least 1 year (mean 30 months). The results of bone-spacer bonding of the 147 implants were evaluated using computerized tomography (CT) scanning. The symptoms significantly improved in 30 patients. No difference in results was detected between patients with cervical spondylosis and those with ossification of posterior longitudinal ligament. Breakage of seven spacers occurred in four patients without causing neck pain or neurological deficits. There were no other HA spacer-related complications. The spacers became rigidly bound to bone in 61% of the cases, and bone regrowth developed around the spacer in 91%. The rate of bone-spacer bonding increased over time, and the CT documented attenuation value (Hounsfield unit) of the spacer adjacent to the bone spacer junction in the group in which union occurred was significantly higher than in the nonunion group. CONCLUSIONS: High-porosity HA spacer-augmented laminoplasty produced good bonding-related results. Bone bonding continued to progress 1 year after surgery, indicating the good osteoconductive capability of high-porosity HA. To avoid breakage of a spacer, a minimum 7-mm distance between spacers is necessary. PMID- 12650398 TI - Implantation of an empty carbon fiber composite frame cage after single-level anterior cervical discectomy in the treatment of cervical disc herniation: preliminary results. AB - OBJECT: The authors sought to evaluate retrospectively the radiological and clinical outcome of anterior cervical discectomy followed by implantation of an empty carbon fiber composite frame cage (CFCF) in the treatment of patients with cervical disc herniation and monoradiculopathy. METHODS: Twenty-five consecutive patients (12 men, 13 women, mean age 45 years) with monoradiculopathy due to cervical disc herniation were treated by anterior cervical discectomy followed by implantation of an empty CFCF cage. On lateral flexion-extension radiographs segmental stability at a mean follow up of 14 months (range 5-31 months) was demonstrated in all 25 patients, and bone fusion was documented in 24 of 25 patients. The mean anterior intervertebral body height was 3.4 mm preoperatively and 3.8 mm at follow up in 20 patients. In these patients the mean segmental angle (angle between lower endplate of lower and upper vertebra) was 0.9 degrees preoperatively and 3.1 degrees at follow up. In the remaining five patients preoperative images were not retrievable. Self-scored neck pain based on a visual analog scale (1, minimum; 10, maximum) changed from a preoperative average of 5.6 to an average of 2 at follow up; radicular pain was reduced from 7.7 to 2.1 postoperatively. Analysis of the SF12 questionnaires showed a significant improvement in both the physical capacity score (preoperative mean 32.4 points; follow up 46 points) and the mental capacity score (preoperative mean 45.8 points; follow up 57.5 points). CONCLUSIONS: Implantation of an empty CFCF cage in the treatment of cervical disc herniation and monoradiculopathy avoids donor site morbidity associated with autologous bone grafting as well as the use of any supplementary material inside the cage. Restoration or maintenance of intervertebral height and thus segmental lordosis and a very high rate of segmental stability and fusion are achieved using this technique. PMID- 12650399 TI - Risk factors for surgical site infection in spinal surgery. AB - OBJECT: The objective of this study was to identify specific independent risk factors for surgical site infections (SSIs) occurring after laminectomy or spinal fusion. METHODS: The authors performed a retrospective case-control study of data obtained in patients between 1996 and 1999 who had undergone laminectomy and/or spinal fusion. Forty-one patients with SSI or meningitis were identified, and data were compared with those acquired in 178 uninfected control patients. Risk factors for SSI were determined using univariate analyses and multivariate logistic regression. The spinal surgery-related SSI rate (incisional and organ space) during the 4-year study period was 2.8%. Independent risk factors for SSI identified by multivariate analysis were postoperative incontinence (odds ratio [OR] 8.2, 95% confidence interval [CI] 2.9-22.8), posterior approach (OR 8.2, 95% CI 2-33.5), procedure for tumor resection (OR 6.2, 95% CI 1.7-22.3), and morbid obesity (OR 5.2, 95% CI 1.9-14.2). In patients with SSI the postoperative hospital length of stay was significantly longer than that in uninfected patients (median 6 and 3 days, respectively; p < 0.001) and were readmitted to the hospital for a median additional 6 days for treatment of their infection. Repeated surgery due to the infection was required in the majority (73%) of infected patients. CONCLUSIONS: Postoperative incontinence, posterior approach, surgery for tumor resection, and morbid obesity were independent risk factors predictive of SSI following spinal surgery. Interventions to reduce the risk for these potentially devastating infections need to be developed. PMID- 12650400 TI - Results of preoperative embolization for metastatic spinal neoplasms. AB - OBJECT: Arterial embolization reduces blood loss in patients undergoing surgery for hypervascular spinal tumors. The objectives of this study were twofold: 1) to evaluate the role of magnetic resonance (MR) imaging in predicting tumor vascularity and 2) to assess the effectiveness of preoperative embolization in devascularizing these tumors. METHODS: Fifty-one patients with metastatic spinal neoplasms underwent angiography, preoperative embolization, and excision of the lesion between 1995 and 2000. The MR imaging studies were correlated with tumor vascularity on angiograms. Embolization was angiographically graded on a five point scale ranging from no embolization (Grade A) to total embolization (Grade E). The embolization grade was correlated with intraoperative blood loss. The mean age was 57 years, the male/female ratio was 1.2:1, and back pain was present in all patients. Metastatic renal cell carcinoma (30 cases) and thoracic spine involvement (33 cases) were most frequent. The positive predictive value of MR imaging in determining tumor vascularity was 77%, whereas the negative predictive value was 21%. Total embolization (Grade E) was achieved in 34 patients. A shared vascular pedicle between a radiculomedullary artery (RMA) and a tumor diminished the likelihood of complete embolization (p = 0.02). Small asymptomatic cerebellar infarctions were demonstrated in two cases. The mean intraoperative blood loss was 2,586 ml. Following Grade D or E embolization, intraoperative bleeding was largely related to unembolized epidural veins. CONCLUSIONS: Tumor histology and MR imaging findings are predictive of hypervascularity, however, hypervascular tumors may not be detected by standard MR imaging sequences. Superselective catheterization permits Grade D or E embolization in 80% of patients. Shared blood supply with an RMA is the most important factor precluding complete embolization. PMID- 12650401 TI - Management of atlantoaxial metastases with posterior occipitocervical stabilization. AB - OBJECT: The treatment of atlantoaxial spinal metastases is complicated by the region's unique biomechanical and anatomical characteristics. Patients most frequently present with pain secondary to instability; neurological deficits are rare. Recently, some authors have performed anterior approaches (transoral or extraoral) for resection of upper cervical metastases. The authors review their experience with a surgical strategy that emphasizes posterior stabilization of the spine and avoidance of poorly tolerated external orthoses such as the rigid cervical collar or halo vest. METHODS: The authors performed a retrospective review of 19 consecutively treated patients with C-1 or C-2 metastases who underwent surgery at The University of Texas M. D. Anderson Cancer Center between 1994 and 2001. Visual analog pain scores were reduced at 1 and 3 months (p < 0.005, Wilcoxon signed-rank test); however, evaluation of pain at 6 months and 1 year was limited by the remaining number of surviving patients. Analgesic medication consumption was unchanged. There were no cases of neurological decline or sudden death secondary to residual or recurrent atlantoaxial disease during the follow-up period. One patient underwent revision of hardware at 11 months. The mean follow-up period was 8 months (range 1-32 months). Median survival determined by Kaplan-Meier analysis was 6.1 months (95% confidence interval 2.99 9.21). CONCLUSIONS: Occipitocervical stabilization provided durable pain relief and preservation of ambulatory status over the remaining life span of patients. Because of the palliative goals of surgery, the authors have not found an indication for anterior-approach tumor resection in these patients. Successful stabilization obviates the need for an external orthosis. PMID- 12650402 TI - Endoscopic anterior cervical foraminotomy for unilateral radiculopathy: anatomical morphometric analysis and preliminary clinical experience. AB - OBJECT: Cervical radiculopathy is typically caused by posterolateral disc herniation or spondylotic foraminal stenosis, either of which may compress the ventral aspect of the nerve root. The authors undertook a study to establish the feasibility of performing an endoscopic approach for anterior cervical foraminotomy (ACFor) in a clinical setting. METHODS: Application of this method on cadavers was conducted to verify the practicability of this technique. The clinical study included 16 patients (eight men and eight women; mean age 46.6 years) all presenting with unilateral radicular symptoms (one at two adjacent ipsilateral levels), which were associated with various degrees of neck pain. Disc herniations and/or uncovertebral osteophytes were confirmed on magnetic resonance imaging and high-resolution computerized tomography scanning. A total of 17 endoscopic ACFors (one two-level procedure) were performed using a rigid glass endoscope (25 degrees angled, 3-mm diameter, 10-mm length) mounted on a tubular retractor. No major surgery-related complications were encountered. During a mean follow-up period of 13.8 months an average absolute improvement of 44% (p > 0.05) in the neck disability index score and of 96% (p > 0.05) in the visual analog scale score for radicular pain (compared with the preoperative score) was observed. During the follow-up period strength improved to normal in 84% and sensory deficit in 80% of the patients. The overall subjective patient satisfaction rate was 87.6%; the return-to-work rate after 4 weeks was 81.4%. CONCLUSIONS: The advantages of endoscopic ACFor include minimial surgical exposure, improved intraoperative visualization, direct decompression of the nerve root, and the preservation of the intervertebral disc and the motion segment. PMID- 12650403 TI - Nuclear factor-kappaB-regulated cyclooxygenase-2 expression in surgery-associated paraspinal muscle injury in rats. AB - OBJECT: Paraspinal muscle injury is a common but neglected complication of posterior spinal surgery. Evidence suggests that surgical retraction places mechanical and oxidative stress on the paraspinal muscles and that inflammation is a major postoperative pathological finding in the muscles. The roles of cyclooxygenase (COX)-2 and nuclear factor (NF)-kappaB in the inflammatory processes after retraction remain to be clarified. METHODS: In the control group, paraspinal muscles were dissected from the spine via a posterior incision and then laterally retracted. Paraspinal muscle specimens were harvested before as well as at designated time points during and after persistent retraction. The time course of NF-kappaB activation was determined by gel shift assay. Expression of COX-2 was examined using Western blot analysis and immunohistochemistry. The severity of inflammation was evaluated based on histopathology and myeloperoxidase (MPO) activity. The NF-kappaB activation was inhibited by the administration of pyrrolidine dithiolcarbamate (PDTC) in the PDTC-treated group. Retraction induced early activation of NF-kappaB in paraspinal muscle cells. The expression of COX-2 could not be detected until 1 day postoperativley, reaching a peak at 3 days. The time course of COX-2 expression correlated with that of inflammatory responses and MPO activity. Pretreatment with PDTC inhibited intraoperative NF-kappaB activation and greatly downregulated postoperative COX-2 expression and inflammation in the muscles. Postinflammation fibrosis was also abolished by PDTC administration. CONCLUSIONS: Both NF-kappaB-regulated COX-2 expression and inflammation play an important role in the pathogenesis of surgery associated paraspinal muscle injury. The therapeutic strategy of NF-kappaB inhibition may be applicable to the prevention of such injury. PMID- 12650404 TI - The transverse process, intertransverse space, and vertebral artery in anterior approaches to the lower cervical spine. AB - OBJECT: The microsurgical anatomy of the C3-6 transverse processes and their relationship to the intertransverse space and vertebral artery (VA) were examined with special attention to the aspect exposed in the anterior surgical approach. METHODS: Ten adult cadaveric spines were examined (magnification levels X 3-40) after perfusion of the arteries and veins with colored silicone. The morphological detail of the transverse process and intertransverse space, the distances between selected surgical landmarks and the VA were measured, and the means and standard deviations were calculated. The osseous changes in the anterior root of the transverse process were classified according to their extent. The transverse processes became smaller, and the anterior intertransverse spaces and the width of the VA exposed in the space increased in size proceeding from caudal to rostral levels, thus exposing the VA to increased risk of injury during procedures at cephalad levels. The distance between the medial border of the longus colli muscle and the VA decreased when proceeding caudally from C2-3 to C4-5 interspaces but began to increase at the level of C5-6. The VA coursed closer to the lateral border of the vertebral body than to the medial border of the anterior tubercle of transverse process. Osseous changes consisting of thinning or defects in the anterior root of the transverse process were observed from C-3 to C-5. The thinning was most prominent in the lower half of the anterior root just above where the VA ascends behind the lower edge of the anterior root. The osseous change may reflect the erosive effect of the VA on the anterior root of the transverse process. CONCLUSIONS: This study provides new information regarding the transverse process and especially the anterior root. An awareness of the thinness and defects in the anterior root of the transverse process and the relationships to the surrounding area will aid in reducing VA injury during anterior approaches to the cervical spine. PMID- 12650405 TI - Suitability of bioresorbable cages for anterior cervical fusion. AB - OBJECT: The authors conducted a pilot study to determine whether a bioresorbable intervertebral fusion device composed of 85/15 polylactide-polyglycolide (PLA PGA) copolymer packed with bone autograft is a suitable alternative to promote arthrodesis after anterior cervical discectomy (ACD) in a caprine model. METHODS: The caprine cervical spine model has been used to evaluate interbody healing and fusion after application of bone grafts and instrumentation. Whether a bioresorbable device is suitable for facilitating intervertebral bone union has not been determined. Twelve goats underwent two-level ACD and fusion; eight received bioresorbable cages packed with autologous bone, and four received autologous bone alone. Goats were maintained without an orthosis and after 12 weeks underwent physical, radiographic, and histological evaluation. Cages had structurally degraded, and two had become extruded. Stable intervertebral union developed in three (19%) of 16 cage-implanted interspaces, and one (14%) of seven bone autograft-implanted interspaces; each was judged manually to be rigid (Grade 2), radiographically to be bridged by new osseous densities (Grade 2), and histologically to have marked new bone formation (Grade 3). A primarily fibrous union, however, stabilized the cage-implanted interspaces, and eight (50%) had developed a 4 to 6-mm foreign body granuloma. These interposed soft tissues were not present in the stable autologous bone-implanted interspace, which had successfully become fused. CONCLUSIONS: Interbody cages composed of 85/15 PLA-PGA copolymer contributed to a stable fibrous union, degraded. and produced granuloma after 12 weeks. Additional evaluations are necessary to determine whether other copolymer mixtures, or other bioresorbable materials, can contribute to an arthodesis without deleterious consequences. PMID- 12650406 TI - Craniovertebral junction fixation with transarticular screws: biomechanical analysis of a novel technique. AB - OBJECT: The authors compared the biomechanical stability resulting from the use of a new technique for occipitoatlantal motion segment fixation with an established method and assessed the additional stability provided by combining the two techniques. METHODS: Specimens were loaded using nonconstraining pure moments while recording the three-dimensional angular movement at occiput (Oc)-C1 and C1-2. Specimens were tested intact and after destabilization and fixation as follows: 1) Oc-C1 transarticular screws plus C1-2 transarticular screws; 2) occipitocervical transarticular (OCTA) plate in which C1-2 transarticular screws attach to a loop from Oc to C-2; and (3) OCTA plate plus Oc-C1 transarticular screws. Occipitoatlantal transarticular screws reduced motion to well within the normal range. The OCTA loop and transarticular screws allowed a very small neutral zone, elastic zone, and range of motion during lateral bending and axial rotation. The transarticular screws, however, were less effective than the OCTA loop in resisting flexion and extension. CONCLUSIONS: Biomechanically, Oc-C1 transarticular screws performed well enough to be considered as an alternative for Oc-C1 fixation, especially when instability at C1-2 is minimal. Techniques for augmenting these screws posteriorly by using a wired bone graft buttress, as is currently undertaken with C1-2 transarticular screws, may be needed for optimal performance. PMID- 12650407 TI - In vitro high-field magnetic resonance imaging-documented anatomy of a fetal myelomeningocele at 20 weeks' gestation. A contribution to the rationale of intrauterine surgical repair of spina bifida. AB - OBJECT: It remains uncertain if closure of a myelomeningocele at midgestation changes the neurological condition at birth in an infant born with spina bifida. The authors conducted a study to provide a detailed analysis of the morphology of the spinal cord with the myelomeningocele at the time fetal surgery usually is performed. METHODS: The myelomeningocele of a 20-week-gestation-age fetus was examined, and data were compared with those obtained in a neurologically intact specimen of the same age. In vitro high-field 9.4-tesla magnetic resonance (MR) microscopy was used to examine the fetal material. High-field MR spectroscopy provided images in the three orthogonal planes with a resolution comparable with low-power optical microscopy. The authors observed that the fetal cord of the myelomeningocele specimen was tapered and tethered at S3-4 while the conus medullaris in the normal fetus reaches L-4. No neurulation defects were noted. The axial MR images clearly revealed the nonfusion of the mesodermal structures. The absence of neurulation defects suggests that at least in some cases of spina bifida the spinal cord initially is well developed but is damaged later on chemically and mechanically. This might be an argument in favor of intrauterine myelomeningocele repair. By 20 weeks' gestation, however, the deformation of the cord inside the myelomeningocele is severe. An optimization of the preoperative assessment by means of MR imaging therefore might be considered a valuable contribution to intrauterine surgery. The in vitro high-field MR microscopic findings of this study could be used as references for clinical intrauterine MR imaging. CONCLUSIONS: The detailed in vitro high-field MR analysis of a 20-week gestation-age fetus with spina bifida demonstrated that an improvement of the preoperative intrauterine imaging should be pursued to detect those cases without neurulation defects and with minimal deformation of the spinal cord. PMID- 12650408 TI - Primary epidural lymphoma. Case report. AB - A case of follicular center cell lymphoma arising in the spinal dura mater is presented. To the authors' knowledge, this is the first case of primary epidural lymphoma in which immunohistochemical and molecular investigations demonstrated a follicular center cell origin. PMID- 12650409 TI - Inflammatory myofibroblastic tumor: a spinal case with aggressive clinical course and ALK overexpression. Case report. AB - The authors report on a case of spinal inflammatory myofibroblastic tumor (IMT) in a 22-year-oldwoman. Neuroradiological features of this intradural extramedullary mass were suggestive of a meningioma or neurinoma. The lesion was easily resected following a T-9 laminectomy. Light microscopy showed a proliferation of spindle cells with prominent nucleoli on a fibrous or edematous background with infiltration of numerous lymphocytes and plasma cells. Some spindle cells immunostained positively for ALK1. This led to the diagnosis of IMT. The patient's postoperative course was complicated by a multifocal local recurrence requiring a second surgery, which was followed by radio- and chemotherapy. The occurrence of IMT in the spinal cord has rarely been reported. In this case, ALK overexpression was associated with early multifocal recurrence. This has been recently reported in this tumor typein other locations. PMID- 12650410 TI - The Wilhelm tell technique for anterior lumbar interbody fusion. Technical note. AB - Experience indicates that stand-alone cages may lack the necessary stability to secure highly unstable motion segments at the lumbosacral junction. The authors have designed a special carbon fiber composite interbody cage that allows additional screw placement in anterior lumbar interbody fusion procedures performed at the lumbosacral junction. PMID- 12650411 TI - Arachnoiditis ossificans. PMID- 12650412 TI - N of 1. PMID- 12650413 TI - Pathophysiology of headache associated with cough in patients with Chiari I malformation. AB - OBJECT: The aim of this study was to evaluate the pathophysiology underlying headache associated with cough in patients with Chiari I tonsillar abnormality. The authors hypothesized that peak intrathecal pressure during coughing is higher in patients with headache aggravated by cough than in patients without or in healthy volunteers. In addition, the authors evaluated the use of intrathecal pressure during cough as a means of assessing obstruction to the free flow of cerebrospinal fluid (CSF) at the craniocervical junction. METHODS: Twenty-six adult patients with Chiari I malformation and syringomyelia, four adult patients with Chiari I malformation without syringomyelia, and 15 healthy volunteers were prospectively studied. Testing before surgery included the following: 1) clinical evaluation for the presence of headache associated with cough; and 2) evaluation of lumbar subarachnoid pressure at rest, during three to five coughs, while performing the Valsalva maneuver, during jugular compression, and after removal of CSF. Patients underwent suboccipital craniectomy, C-1 laminectomy, and duraplasty. Testing was repeated 6 months after surgery. CONCLUSIONS: Peak intrathecal pressures during cough and at baseline were elevated in patients with headache associated with cough compared with either patients without headache or healthy volunteers. After surgery, intrathecal pressures during cough were significantly lower than preoperative values and headache aggravated by cough was resolved partially or completely. Headache linked to coughing in patients with Chiari I malformation is associated with sudden increased intrathecal pressure caused by obstruction to the free flow of CSF in the subarachnoid space. PMID- 12650414 TI - Improvement in brainstem auditory evoked potentials after suboccipital decompression in patients with chiari I malformations. AB - OBJECT: The optimal treatment for patients with symptoms related to Chiari I malformation remains controversial. Although a suboccipital decompression with duraplasty is most commonly performed, there may be a subset of patients who improve in response to bone decompression alone. In an initial attempt to identify such patients, we performed a continuous study of intraoperative brainstem auditory evoked potentials (BAEPs) in patients undergoing a standard decompression with duraplasty and compared conduction times at three different time points: 1) baseline while the patient is supine (before positioning); 2) immediately after opening of the bone and release of the atlantooccipital membrane (that is, the dural band); and 3) after opening of the dura mater. METHODS: Eleven children and young adults (mean age 9.8 years) with symptoms related to Chiari I malformation underwent suboccipital decompression and duraplasty with intraoperative monitoring of BAEPs and somatosensory evoked potentials (SSEPs). Six patients (55%) had associated syringomyelia. At baseline, the I to V interpeak latency (IPL) for both sides (total 21 BAEPs) was 4.19 +/- 0.22 msec (mean +/- standard deviation). After complete bone decompression and before the dura mater was opened, the I to V IPL decreased to 4.03 +/- 0.25 msec (p = 0.0005). When the dura was opened, however, no further decrease in the I to V IPL was detected (4.03 +/- 0.25 msec; p = 0.6). The SSEPs remained stable throughout the procedure. CONCLUSIONS: In children and young adults undergoing suboccipital decompression with duraplasty for Chiari I malformation, the vast majority of improvement in conduction through the brainstem occurs after bone decompressionand division of the atlantooccipital membrane, rather than after opening of the dura. Additional studies are needed to establish whether the improvement seen with BAEP monitoring during bone decompression will predict long term clinical improvement in these patients. PMID- 12650415 TI - Localization of language function in children: results of electrical stimulation mapping. AB - OBJECT: The authors examined the localization of language sites and the frequency of naming errors at these sites in a population of children undergoing electrical stimulation mapping during surgeries in which epileptic foci and dominant hemisphere neoplasms were resected. The frequency with which essential language sites were found (that is, "the frequency of language sites") in children was compared with that of a population of adults who had undergone this procedure, to assess the relationship of age to the distribution of essential areas for language. METHODS: The results of electrical stimulation mapping to determine sites of naming and speech arrest in 26 children ranging in ages from 4 to 16 years are presented in this report. Mapping was performed in the intraoperative setting in eight patients and in the extraoperative setting, by stimulation across a subdural grid, in 18 patients. The frequency and distribution of essential language areas were analyzed in populations of different ages and according to the method used to obtain the map. Considerable variability was found in the localization of language sites. When the language site distribution in pediatric patients was compared with the language site distribution found previously in a population of patients older than 16 years of age, a relative paucity of language sites was found in all perisylvian cortices in the younger age group. This relationship was also found within the group of patients 16 years of age and younger, when segregated into two groups: those patients 8 years of age or younger, and those patients between 9 and 16 years of age. These findings are relevant to theories of the intrahemispheric organization of the cortex devoted to language function. CONCLUSIONS: The differences found between groups of younger and older patients in the frequencies of sites where stimulation produces naming errors was identified suggests the possibility that, with advancing age, maturational processes contribute new foci of cortex essential for language. PMID- 12650416 TI - Clinical experience with the use of a shunt with an adjustable valve in children with hydrocephalus. AB - OBJECT: The goal of this study was to assess the value of making adjustments in the opening pressure of a shunt valve and to determine shunt survival in children and young adults in whom an adjustable valve was used to manage cerebrospinal fluid drainage. METHODS: The authors conducted a single-center retrospective study of 158 children and young adults who had received 199 Codman Hakim programmable valves (noninvasively adjustable to settings in the range of 30-200 mm H2O). The mean age at which the patients underwent shunt implantation was 4.4 years (median 0.4 years, maximum 18 years); 94 patients were younger than 2 years of age, including 14 patients with a gestational age younger than 38 weeks at the time of implantation. In 84 (53.2%) of the 158 patients, valve pressure adjustment was required at least once (mean 1.3 times, maximum 16 times). Among the 202 adjustments made in patients the reason for adjustment was underdrainage in 74 adjustments (36.6%) and overdrainage in 119 (58.9%). The clinical status of the patient improved after 121 (69.1%) of 175 adjustments and after 58 (73.4%) of 79 minor adjustments (less than or equal to +/- 20 mm H2O). The shunt infection rate was 13 (10.9%) of 119 patients. Shunt survival was 60.5% at 1 year, 47.1% at 2 years, and 43.9% at 3 years of follow-up review. CONCLUSIONS: Adjustment of the valve's opening pressure further improves outcome in pediatric patients. PMID- 12650417 TI - Symptom-based assessment of the severity of a concussion. AB - OBJECT: Current grading systems of concussion and return-to-play guidelines have little empirical support. The authors therefore examined the relationships of the characteristics and symptoms of concussion and the history of concussion to three indicators of concussion severity-number of immediate symptoms, number of symptoms at the initial follow-up examination, and duration of symptoms--to establish an empirical basis for grading concussions. METHODS: Forty-seven athletes who sustained concussions were administered alternate forms of an Internet-based neurocognitive test until their performances were within normal limits relative to baseline levels. Assessments of observer-reported and self reported symptoms at the sideline of the playing field on the day of injury, and at follow-up examinations were also obtained as part of a comprehensive concussion management protocol. Although loss of consciousness (LOC) was a useful indicator of the initial severity of the injury, it did not correlate with other indices of concussion severity, including duration of symptoms. Athletes reporting memory problems at follow-up examinations had significantly more symptoms in general, longer durations of those symptoms, and significant decreases in scores on neurocognitive tests administered approximately 48 hours postinjury. This decline of scores on neurocognitive testing was significantly associated with an increased duration of symptoms. A history of concussion was unrelated to the number and duration of symptoms. CONCLUSIONS: This paper represents the first documentation of empirically derived indicators of the clinical course of postconcussion symptom resolution. Self-reported memory problems apparent 24 hours postconcussion were robust indicators of the severity of sports-related concussion and should be a primary consideration in determining an athlete's readiness to return to competition. A decline on neurocognitive testing was the only objective measure significantly related to the duration of symptoms. Neither a brief LOC nor a history of concussion was a useful predictor of the duration of postconcussion symptoms. PMID- 12650418 TI - Framework for bioethical assessment of an article on therapy. AB - OBJECT: Frameworks for scientific assessment of articles on therapy published in the medical literature have become available and will likely enhance the quality of medical research that is published in peer-reviewed journals. Comprehensive frameworks do not exist for the assessment of bioethical issues pertaining to research on human volunteers. METHODS: The authors have developed a framework consisting of ethical dimensions or questions that they suggest should be applied to assess the bioethical integrity of articles on therapy. Thirteen questions were developed and discussed in the context of current bioethical principles, and examples were applied where possible. CONCLUSIONS: The simple framework the authors have developed offers a method to assess key bioethical issues surrounding an article on therapy and probably defines the minimum standard to which such articles should be held. Many ethical questions cannot yet be answered based on available information or bioethical theories. The authors are not suggesting that their framework is comprehensive; refinements and individualization of it to fit specific studies are probably required by each clinician-researcher who designs a therapy trial and reports its results. PMID- 12650419 TI - Endovascular therapy for stenosis of the petrous or cavernous portion of the internal carotid artery: percutaneous transluminal angioplasty compared with stent placement. AB - OBJECT: The effects of percutaneous transluminal angioplasty (PTA) and stent placement for stenosis of the petrous or cavernous portion of the internal carotid artery (ICA) were compared. METHODS: Twenty-four patients with symptomatic, greater than 60% stenosis of the petrous or cavernous portion of the ICA were treated using PTA or stent placement; 15 were treated with PTA and nine with stent insertion. Initial and follow-up results (> 3 months posttreatment) were compared in each group. Stenotic portions of the ICA were successfully opened in 13 of 15 patients in the PTA group, and in all nine patients in the stent-treated group. In one case in the PTA group stent delivery was attempted; however, the device could not pass through the vessel's tortuous curve, and PTA alone was performed in this case. Postoperatively, the mean stenotic ratio decreased from 72.1 to 29.6% in the PTA group, and from 75.6 to 2.2% in the stent treated group. In four patients in the PTA group, stenoses greater than 50% were demonstrated on follow-up angiography performed at 3 to 6 months after PTA. In the stent-treated group, no restenosis was encountered, although in one case acute occlusion of the stent occurred; the device was recanalized with PTA and infusion of tissue plasminogen activator. This case was the only one of the 24 in which any neurological deficits related to the endovascular procedure occurred. Stent placement brought a greater gain in diameter than did PTA at the initial and late follow-up period; this gain was statistically significant. CONCLUSIONS: Stent placement is more effective than PTA for stenosis of the petrous or cavernous portion of the ICA from the viewpoint of initial and late gain in diameter. PMID- 12650420 TI - Use of a wedged microcatheter for curative transarterial embolization of complex intracranial dural arteriovenous fistulas: indications, endovascular technique, and outcome in 21 patients. AB - OBJECT: The aim of this study was to describe the application of a novel transarterial approach to curative embolization of complex intracranial dural arteriovenous fistulas (DAVFs). This technique is particularly useful in patients harboring high-grade DAVFs with direct cortical venous drainage or for whom transvenous coil embolization is not possible because of limited sinus venous access to the fistula site due to thrombosis or stenotic changes. METHODS: Twenty three DAVFs in 21 patients were treated using a transarterial N-butyl cyanoacrylate (NBCA) embolization technique with the aid of a wedged catheter. In all patients, definitive treatment involved two critical steps: 1) a microcatheter was wedged within a feeding artery, establishing flow-arrest conditions within the catheterized vessel distal to the microcatheter tip; and 2) NBCA was injected under these resultant flow-arrest conditions across the pathological arteriovenous connection and into the immediate draining venous apparatus, definitively occluding the fistula. Patient data were collected in a retrospective manner by reviewing office and inpatient charts and embolization reports, and by directly analyzing all procedural and diagnostic angiograms. Eight patients presented with the principal complaint of tinnitus/bruit, five with intracranial hemorrhage, four with cavrnous sinus syndrome, and one each with seizures, ataxia, visual field loss, and hiccups. The parent (recipient) venous structure of the DAVFs in this study included 11 leptomeningeal veins, eight transverse/sigmoid sinuses, three cavernous sinuses, and one sphenoparietal sinus. The NBCA permeated the arteriovenous shunt, perifistulous network, and proximal draining vein in all DAVFs. Occlusion was confirmed on postembolization angiography studies. No complication occurred in any patient in this series. There has been no recurrence during a mean follow up of 18.7 months (range 2-46 months). CONCLUSIONS: Transarterial NBCA embolization with the aid of a wedged catheter in flow-arrest conditions is a safe and an effective treatment for intracranial DAVFs. PMID- 12650421 TI - Intraoperative monitoring of blood flow insufficiency in the anterior choroidal artery during aneurysm surgery. AB - OBJECT: The lack of a specified intraoperative method for monitoring anterior choroidal artery (AChA) blood flow insufficiency (BFI) led the authors to devise a method for checking the BFI in this artery during aneurysm surgery. To this end, the authors relied on the intraoperative motor evoked potentials (MEPs) elicited by electrical stimulation of the hand motor cortex. METHODS: The study population consisted of 108 patients with internal carotid artery (ICA) aneurysms who underwent surgery via a standard frontotemporal craniotomy. After the dura mater had been opened, a grid electrode strip with 16 small electrodes was inserted subdurally into the hand motor cortex from the edge of the craniotomy. To check BFI in the AChA, the hand motor cortex was stimulated at an intensity level between 10 and 18 mA. The MEPs were successfully recorded from the contralateral thenar muscles in all 108 patients. There was no postoperativemotor paresis in 88 patients in whom the MEPs remained unchanged during the performance of various surgical maneuvers. Among the other 20 patients, 19 manifested transient MEP changes, but 15 of those patients experienced no postoperative motor paresis. In four patients who exhibited transient MEP changes, either after aneurysm clipping or during temporary occlusion of the ICA and/or AChA, hemiparesis occurred postoperatively but disappeared within 24 hours. In one patient with an ICA-posterior communicating artery aneurysm, the MEP disappeared and did not reappear by the time of dural closure. Severe hemiplegia developed in this patient and a computerized tomography scan obtained postoperatively revealed a new low-density area in the internal capsule. CONCLUSIONS: The findings of this study suggest that the monitoring method that is introduced here is safe and reliable for detecting intraoperative BFI in the AChA. PMID- 12650422 TI - Monitoring of brain tissue oxygenation during aneurysm surgery: prediction of procedure-related ischemic events. AB - OBJECT: The aim of this study was to evaluate the feasibility of monitoring brain tissue oxygenation (PO2) during aneurysm surgery for the detection of procedure related ischemia. METHODS: Between 1997 and 1998, PO2 was monitored prospectively in a cohort of 40 patients (42 recordings) during aneurysm surgery in the anterior circulation within the vascular territory of the aneurysm-bearing artery. The position of the probe used to measure oxygenation levels was verified on computerized tomography (CT) scanning on the 1st postoperative day. Because of the mislocation of one probe and the malfunction of another, data from only 38 patients (40 recordings) were suitable for analysis. Relative changes from baseline to absolute nadir values of intraoperative PO2 were correlated with simultaneously recorded somatosensory evoked potentials (SSEPs), and cardiovascular and ventilatory parameters. The frequency of ischemic events was evaluated with the aid of CT on the 1st postoperative day as a substitute parameter for intraoperative ischemia. Clinical outcome was evaluated 30 days postoperatively based on the Glasgow Outcome Scale. Except for three, all patients underwent surgery for treatment of a symptomatic aneurysm. Mean baseline PO2 was 23.9 mm Hg (range 2-67.2 mm Hg) before clip application. A relative decrease in PO2 (20% decrease in value compared with baseline) occurred in 12 patients and was a sensitive indicator for the risk of ischemia during temporary arterial occlusion, but was less predictive of nonocclusive ischemia (sensitivity 0.5; positive predictive value [PPV] 0.42; p > 0.05). Results of receiver operating characteristic analysis demonstrated a postclipping PO2 nadir of 15 mm Hg as a dichotomizing threshold for the prediction of ischemia. This threshold rendered an improved sensitivity (0.9) and PPV (0.56) for procedure-related ischemia (p = 0.0003). The results of utility analysis revealed this monitoring parameter to be clinically diagnostic. Only PO2 monitoring, and not SSEP at the tibial nerve, was predictive of ischemia within the anterior cerebral artery territory. CONCLUSIONS: Using 15 mm Hg as a dichotomizing threshold, intraoperative PO2 monitoring enables one to identify patients at risk for procedure-related ischemia during aneurysm surgery and surpasses SSEP monitoring. This newly defined threshold based on intraoperative PO2 monitoring provides a basis for studies on treatments for procedure-related ischemia during aneurysm surgery. PMID- 12650423 TI - Use of the peak troponin value to differentiate myocardial infarction from reversible neurogenic left ventricular dysfunction associated with aneurysmal subarachnoid hemorrhage. AB - OBJECT: Differentiating myocardial infarction (MI) from reversible neurogenic left ventricular dysfunction (stunned myocardium [SM]) associated with aneurysmal subarachnoid hemorrhage (SAH) is critical for early surgical intervention. The authors hypothesized that the cardiac troponin (cTn) trend and/or echocardiogram could be used to differentiate between the two entities. METHODS: A retrospective study was conducted for the period between 1995 and 2000. All patients included in the study met the following criteria: 1) no history of cardiac problems; 2) new onset of abnormal cardiac function (ejection fraction [EF] < 40% on echocardiograms); 3) serial cardiac markers (cTn and creatine kinase MB isoform [CK-MB]); 4) surgical intervention for their aneurysm; and 5) cardiac output monitoring either by repeated echocardiograms or invasive hemodynamic monitoring during the first 4 days post-SAH when the patients were euvolemic. Of the 350 patients with SAH, 10 (2.9%) had severe cardiac dysfunction. Of those 10, six were women and four were men. The patients' mean age was 53.5 years (range 29-75 years) and their SAH was classified as Hunt and Hess Grade III or IV. Aneurysm distribution was as follows: basilar artery tip (four); anterior communicating artery (two); middle cerebral artery (one); posterior communicating artery (two); and posterior inferior cerebellar artery (one). The mean EFonset was 33%. The changes on echocardiograms in these patients did not match the findings on electrocardiograms (EKGs). Within 4.5 days, dramatic improvement was seen in cardiac output (from 4.93 +/- 1.16 L/minute to 7.74 +/- 0.88 L/minute). Compared with historical controls in whom there were similar levels of left ventricular dysfunction after MI, there was no difference in peak CK-MB. A 10-fold difference, however, was noted in cTn values (0.22 +/- 0.25 ng/ml; control 2.8 ng/ml; p < 0.001). CONCLUSIONS: The authors determined the following: 1) that the CK-MB trend does not allow differentiation between SM and MI; 2) that echocardiograms revealing significant inconsistencies with EKGs are indicative of SM; and 3) that cTn values less than 2.8 ng/ml in patients with EFs less than 40% are consistent with SM. PMID- 12650424 TI - Comparison between perimesencephalic nonaneurysmal subarachnoid hemorrhage and subarachnoid hemorrhage caused by posterior circulation aneurysms. AB - OBJECT: Some authors have questioned the need to perform cerebral angiography in patients presenting with a benign clinical picture and a perimesencephalic pattern of subarachnoid hemorrhage (SAH) on initial computerized tomography (CT) scans, because the low probability of finding an aneurysm does not justify exposing patients to the risks of angiography. It has been stated, however, that ruptured posterior circulation aneurysms may present with a perimesencephalic SAH pattern in up to 10% of cases. The aim of the present study was twofold: to define the frequency of the perimesencephalic SAH pattern in the setting of ruptured posterior fossa aneurysms, and to determine whether this clinical syndrome and pattern of bleeding could be reliably and definitely distinguished from that of aneurysmal SAH. METHODS: Twenty-eight patients with ruptured posterior circulation aneurysms and 44 with nonaneurysmal perimesencephalic SAH were selected from a series of 408 consecutive patients with spontaneous SAH admitted to the authors' institution. The admission unenhanced CT scans were evaluated by a neuroradiologist in a blinded fashion and classified as revealing a perimesencephalic SAH or a nonperimesencephalic pattern of bleeding. Of the 28 patients with posterior circulation aneurysms, five whose grade was I according to the World Federation of Neurosurgical Societies scale were classified as having a perimesencephalic SAH pattern on the initial CT scan. The data show that the likelihood of finding an aneurysm on angiographic studies obtained in a patient with a perimesencephalic SAH pattern is 8.9%. Conversely, ruptured aneurysms of the posterior circulation present with an early perimesencephalic SAH pattern in 16.6% of cases. CONCLUSIONS: This study supports the impression that there is no completely sensitive and specific CT pattern for a nonaneurysmal SAH. In addition, the authors believe that there is no specific clinical syndrome that can differentiate patients who have a perimesencephalic SAH pattern caused by an aneurysm from those without aneurysms. Digital subtraction angiography continues to be the gold standard for the diagnosis of cerebral aneurysms and should be performed even in patients who have the characteristic perimesencephalic SAH pattern on admission CT scans. PMID- 12650425 TI - Three-dimensional angiography for radiosurgical treatment planning for arteriovenous malformations. AB - OBJECT: Radiosurgical treatment of a cerebral arteriovenous malformation (AVM) requires the precise definition of the nidus of the lesion in stereotactic space. This cannot be accomplished using simple stereotactic angiography. but requires a combination of stereotactic biplanar angiographic images and stereotactic contrast-enhanced computerized tomography (CT) scans. In the present study the authors describe a method in which three-dimensional (3D) rotational angiography is integrated into stereotactic space to aid treatment planning for radiosurgery. METHODS: Twenty patients harboring AVMs underwent treatment planning prior to linear accelerator radiosurgery. Planning involved the acquisition of two different data sets, one of which was obtained using the standard method (a combination of biplanar stereotactic angiography with stereotactic CT scanning), and the other, which was procured using a new technique (nonstereotactic 3D rotational angiography combined with stereotactic CT scanning by a procedure of image fusion). The treatment plan that was developed using the new method was compared with that developed using the standard one. For each patient the number of isocenters and the dimension of selected collimators were the same, based on the information supplied in both methods. Target coordinates were modified in only five cases and by a limited amount (mean 0.7 mm, range 0.3-1 mm). CONCLUSIONS: The new imaging modality offers an easier and more immediate interpretation of 3D data, while maintaining the same accuracy in target definition as that provided by the standard technique. Moreover, the new method has the advantage of using nonstereotactic 3D angiography, which can be performed at a different site and a different time with respect to the irradiation procedure. PMID- 12650426 TI - In vivo model of intracranial stent implantation: a pilot study to examine the histological response of cerebral vessels after randomized implantation of heparin-coated and uncoated endoluminal stents in a blinded fashion. AB - OBJECT: No animal model currently exists for the examination of time-dependent histological changes occurring in intracranial vessels after endoluminal stent placement. The authors' goal was to develop a reproducible in vivo model of stent implantation in intracranial vessels in dogs that was capable of demonstrating stent-related vascular changes after the implantation of coated and uncoated devices. METHODS: The authors implanted heparin-coated or uncoated stents in the basilar arteries (BAs) of 11 mongrel dogs. In a 12th animal, one coated stent was implanted in the BA and a second uncoated one was implanted in the distalanterior spinal artery. All the devices were oversized to induce intimal injury. Surviving animals were observed for 12 weeks, after which they underwent repeated angiography before planned death and removal of the brain. Histological studies and computer-assisted morphometric analyses were conducted on stent-treated and untreated sections of the BAs to assess the percentage of stenosis, neointimal proliferation, vessel injury, and inflammation. Perforating vessels partially covered by stent struts ("jailing") were studied for evidence of stenosis or occlusion. The pathologist, interventionists, histopathologist, histopathology technicians, and radiologist were blinded to the stent type. Seven stents (three uncoated and four coated) were removed from the six animals that were observed during the follow-up period. The mean neointimal proliferation was 0.42 mm2 in the group treated with uncoated stents and 0.18 mm2 in the group treated with heparin-coated devices (p = 0.04). Neointimal thickness was significantly increased in the group with uncoated stents (p = 0.04). The mean percentage of occlusion was less (12%) in the group with heparin-coated stents, compared with 22% in the group with uncoated devices (p = 0.07). When comparing results between the heparin-coated and uncoated devices implanted in the five animals that received a single stent only, greater differences (indicating a benefit from heparin-coated stents) were observed in neointimal area (p = 0.009), neointima/media ratio (p = 0.001), neointimal thickness (p = 0.002), and percentage of occlusion (p = 0.009). All brainstem perforating vessels covered by stent struts remained patent. CONCLUSIONS: This in vivo intracranial stent model was developed to assess proliferative and inflammatory responses to endoluminal stent implantation in the cerebrovasculature. The results indicate that a lower percentage of occlusion occurs 12 weeks after implantation of heparin-coated compared with uncoated stents. PMID- 12650427 TI - Adenosine-induced modulation of excitatory amino acid transport across isolated brain arterioles. AB - OBJECT: Excitatory amino acid (EAA) uptake by neurons and glia acts synergistically with stereoselective transport across the blood-brain barrier (BBB) to maintain EAA homeostasis in the brain. The endogenous neuroprotectant adenosine counteracts many aspects of excitotoxicity by increasing cerebral blood flow and by producing pre- and postsynaptic actions on neurons. In the present study, the authors explored the effect of adenosine on EAA transport across the BBB. METHODS: The effects of adenosine on the permeability of the BBB and transport of aspartate and glutamate across the BBB were studied in a well characterized isolated penetrating cerebral arteriole preparation suitable for simultaneous investigations of changes in diameter and permeability. At concentrations within the physiological to low pathophysiological range (10(-7) 10(-6) M), the net vectorial transport of [3H]L-glutamate or [3H]L-aspartate from blood to brain was significantly attenuated, whereas there was no effect of adenosine on paracellular BBB permeability to [14C]sucrose or [3H]D-aspartate. With higher concentrations of adenosine (10(-4) M and 10(-3) M) the net vectorial transport of [3H]L-glutamate and [3H]Laspartate returned toward baseline. At 10( 3) M, the permeability to [14C]sucrose was significantly altered, indicating a breakdown in the BBB. The effect of adenosine (10(-6) M) was blocked by theophylline, a blocker of the A1 and A2 receptors of adenosine. CONCLUSIONS: Adenosine-mediated modulation of glutamate and aspartate transport across the BBB is a novel physiological finding. PMID- 12650428 TI - Effects of the Ca++-permeable nonselective cation channel blocker LOE 908 on subarachnoid hemorrhage-induced vasospasm in the basilar artery in rabbits. AB - OBJECT: The Ca++ influx into vascular smooth-muscle cells (VSMCs) plays a fundamental role in the development and chronic effects of vasospasm after subarachnoid hemorrhage (SAH). The Ca++-permeable nonselective cation channels (NSCCs) are activated by several endothelium-derived constricting factors such as endothelin 1 (ET-1) and thromboxane A2. Moreover, the receptor-operated Ca++ channel blocker LOE 908 inhibits ET-1-induced extracellular Ca++ influx via NSCCs in the VSMCs of the basilar artery (BA) and the NSCC-dependent part of ET-1 induced vasoconstriction of BA rings. The purpose of the present study was to evaluate the in vivo role of LOE 908 on SAH-induced vasospasm. METHODS: Forty-two Japanese white rabbits were assigned to seven groups. Treatment groups consisted of the following: 1) control rabbits without SAH that received a cisternal injection of saline; 2) rabbits with SAH that were subjected to the intravenous administration of saline; 3 through 6) rabbits with SAH that underwent the intravenous administration of 0.01. 0.1, 1, or 10 mg/kg LOE 908, respectively; and 7) rabbits without SAH that underwent the intravenous administration of 10 mg/kg LOE 908. Autologous blood was injected into the cisterna magna. The caliber of the BA was measured on angiographic studies before and after the cisternal injection of autologous blood. The intravenous injection of LOE 908 inhibited the magnitude of an SAH-induced vasosapsm. In addition, the concentration of LOE 908 required to relax vasospasm (1 mg/kg) correlated with that required to block Ca++ influx into VSMCs. CONCLUSIONS: The Ca++ channel blocker LOE 908 may inhibit the magnitude of an SAH-induced vasospasm by blocking the influx of Ca++ through NSCCs in rabbit BAs. Blocking the NSCCs may represent a new treatment for cerebral vasospasm after SAH. PMID- 12650429 TI - Regulation of aquaporin-4 in a traumatic brain injury model in rats. AB - OBJECT: Aquaporin-4 (AQP4) plays a significant role in the regulation of brain water homeostasis. In this study the authors investigated the regulation of AQP4 following a focal cortical contusion injury in rats. METHODS: Thirty-three adult male Wistar rats received a focal cortical contusion of the parietal cortex. An additional nine rats underwent a craniectomy, but no trauma was inflicted (sham injury). Animals were killed 1, 4, and 24 hours later. The rat brains were examined for water content by comparing the wet and dry weights of each hemisphere. Aquaporin-4 messenger (m)RNA was measured by reverse transcription polymerase chain reaction. A ratio of AQP4 mRNA expression in the lesioned hemisphere compared with that in the contralateral control hemisphere was calculated for each animal at the injury site (parietal cortex) and at sites adjacent to (occipital cortex) and distant from the injury (frontal pole cortex). Brain edema was significantly increased at the injury site. The expression of AQP4 mRNA was significantly increased at the injury site, significantly decreased adjacent to the injury site, and not significantly different at a site distant from the injury. The magnitude of AQP4 mRNA upregulation at the injured parietal cortex correlated with the degree of downregulation in the adjacent occipital cortex. CONCLUSIONS: Data from this study demonstrate that an upregulation of AQP4 occurs at the site of traumatic brain injury and that a downregulation of this molecule occurs adjacent to the site of injury. Understanding the physiology of AQP4 and its regulation following brain injury may allow for the development of novel treatments for cerebral edema that accompanies head injury. PMID- 12650430 TI - Camptothecin analogs in malignant gliomas: comparative analysis and characterization. AB - OBJECT: The authors compared and characterized several new classes of camptothecin (CPT) analogs (a total of 22 drugs) directed against human and murine glioma cell lines in vitro, trying to identify CPT analogs that can be used for local therapy in future clinical trials. Camptothecin is a naturally occurring alkaloid that inhibits the DNA-replicating enzyme topoisomerase I. Moreover, CPT and its analogs have shown promising antitumor activity against both systemic and intracranial neoplasms. Because the CPTs have poor bioavailability and are unable to cross the blood-brain barrier, they may best be delivered to the central nervous system by polymers. The authors have previously shown that local delivery of Na-CPT by implantable polymers prolongs survival in a rat intracranial glioma model. In recent years, a number of newly synthesized CPT analogs have been developed that exhibit more potency and stability than Na CPT. METHODS: Cytotoxicities of the drugs were tested using modified clonogenic and monotetrazolium assays in three glioma cell lines. A potassium chloride sodium dodecyl sulfate coprecipitation assay was used to determine the frequency of drug-stabilized cleavable complexes. Of the CPT analogs analyzed, the 10,11 methylenedioxy (MD) class consistently demonstrated the greatest cytotoxicity. Three of these analogs, 10,11-MD-20(RS)-CPT, 10,11-MD-20(S)-CPT-glycinate ester (Gly).HCl, and 9-amino-10,11-MD-20(S)-CPT-Gly, exhibit significantly greater antiproliferative activities than CPT, Na-CPT, or 1,3-bis(2-chloroethyl)-1 nitrosourea (BCNU) against all three glioma cell lines. In addition, the 10,11-MD 20(RS)-CPT analog induces more cleavable complexes than Na-CPT at every concentration. CONCLUSIONS: The increased potency and greater stability of CPT analogs hold promise for more effective local antitumor treatments against malignant intracranial brain tumors. The greater cytotoxicity of 10,11-MD CPTs in comparison with other CPT analogs as well as CPT, BCNU, or Na-CPT, may present an ideal candidate drug class for development against both primary and metastatic brain tumors. PMID- 12650431 TI - Stretching and breaking characteristics of cerebrospinal fluid shunt tubing. AB - OBJECT: Cerebrospinal fluid (CSF) shunt system malfunction due to silastic tubing fracture necessitates revision surgery in shunt-dependent individuals. The goal of this study was to examine the mechanical stretching and breaking characteristics of new and used CSF shunt tubing catheters to determine if any inherent physical properties predispose the tubing to fracture. METHODS: Fifty millimeter segments of new and retrieved (used) CSF shunt tubing were stretched to 120 mm in a hydraulic press to determine modulus values (modulus = stress/strain) and to measure permanent tubing deformation imparted by the applied stress and strain. Similar 50-mm tubing segments were also stretched in an electromechanical material testing system until fracture occurred; the force and strain needed to break the tubing was recorded at the time of failure. The results demonstrate that shunt tubing with a greater cross-sectional area requires greater force to fracture, and that catheters become weaker the longer they are implanted. Barium-impregnated shunt tubing, compared with translucent tubing. appears to require less applied stress and strain to break and may fracture more easily in vivo. The variety of modulus values obtained for the new catheters tested indicates that the various companies may be using materials of different quality in tubing manufacture. CONCLUSIONS: A CSF shunt catheter design that incorporates tubing with a greater cross-sectional area may lead to fewer fractures of indwelling catheters and a reduction in shunt revision surgery. PMID- 12650432 TI - Convective distribution of macromolecules in the primate brain demonstrated using computerized tomography and magnetic resonance imaging. AB - OBJECT: Convection-enhanced delivery (CED), the delivery and distribution of drugs by the slow bulk movement of fluid in the extracellular space, allows delivery of therapeutic agents to large volumes of the brain at relatively uniform concentrations. This mode of drug delivery offers great potential for the treatment of many neurological disorders, including brain tumors, neurodegenerative diseases, and seizure disorders. An analysis of the treatment efficacy and toxicity of this approach requires confirmation that the infusion is distributed to the targeted region and that the drug concentrations are in the therapeutic range. METHODS: To confirm accurate delivery of therapeutic agents during CED and to monitor the extent of infusion in real time, albumin-linked surrogate tracers that are visible on images obtained using noninvasive techniques (iopanoic acid [IPA] for computerized tomography [CT] and Gd diethylenetriamine pentaacetic acid for magnetic resonance [MR] imaging) were developed and investigated for their usefulness as surrogate tracers during convective distribution of a macromolecule. The authors infused albumin-linked tracers into the cerebral hemispheres of monkeys and measured the volumes of distribution by using CT and MR imaging. The distribution volumes measured by imaging were compared with tissue volumes measured using quantitative autoradiography with [14C]bovine serum albumin coinfused with the surrogate tracer. For in vivo determination of tracer concentration, the authors examined the correlation between the concentration of the tracer in brain homogenate standards and CT Hounsfield units. They also investigated the long-term effects of the surrogate tracer for CT scanning, IPA-albumin, on animal behavior, the histological characteristics of the tissue, and parenchymal toxicity after cerebral infusion. CONCLUSIONS: Distribution of a macromolecule to clinically significant volumes in the brain is possible using convection. The spatial dimensions of the tissue distribution can be accurately defined in vivo during infusion by using surrogate tracers and conventional imaging techniques, and it is expected that it will be possible to determine local concentrations of surrogate tracers in voxels of tissue in vivo by using CT scanning. Use of imaging surrogate tracers is a practical, safe, and essential tool for establishing treatment volumes during high-flow interstitial microinfusion of the central nervous system. PMID- 12650433 TI - Delayed repletion of O6-methylguanine-DNA methyltransferase resulting in failure to protect the human glioblastoma cell line SF767 from temozolomide-induced cytotoxicity. AB - OBJECT: Temozolomide (TMZ)-induced O6-methylguanine (MG) DNA lesions, if not removed by MG-DNA methyltransferase (MGMT), mispair with thymine, trigger rounds of futile mismatch repair (MMR), and in glioma cells lead to prolonged G2-M arrest and ultimately cell death. Depletion of MGMT by O6-benzylguanine (BG) sensitizes tumor cells to TMZ, and this combination is currently used in clinical trials. The use of the TMZ+BG combination in gliomas, however, is complicated by the prolonged TMZ-induced G2-M arrest, which may delay activation of poorly defined cell death pathways and allow for MGMT repletion and reversal of toxicity. METHODS: To address these issues, the actions of TMZ were monitored in DNA MMR-proficient SF767 glioma cells depleted of MGMT by BG, and in cells in which BG was removed at various times after TMZ exposure. In MGMT-depleted cells, TMZ exposure led to DNA single-strand breaks and phosphorylation of cdc2, followed by G2-M arrest, induction of p53/p21, and DNA double-strand breaks. Although DNA single-strand breaks, phosphorylation of cdc2, and G2-M arrest could be reversed by repletion of MGMT up to 5 days after TMZ exposure, TMZ-induced cytotoxicity could only be prevented if MGMT was replenished within 24 hours of the onset of G2-M arrest, and before the creation of DNA double-strand breaks. CONCLUSIONS: These results indicate that although SF767 glioma cells undergo a prolonged G2-M arrest in response to TMZ, their ability to escape TMZ-induced cytotoxicity by MGMT repletion is limited to an approximately 24-hour period after the onset of G2-M arrest. PMID- 12650434 TI - Experimental appraisal of the lack of antitumor natural killer cell-mediated immunosurveillance in response to lymphomas growing in the mouse brain. AB - OBJECT: Natural killer (NK) cell-mediated immunosurveillance in the brain is currently obscure, in contrast with the intracerebral immune reaction of cytotoxic T lymphocytes (CTLs) to tumor cells. The goal of this study, in which a controlled tumor model was used, was to investigate a relationship between NK cells and major histocompatibility complex (MHC) Class I gene expression in intracerebral tumor-bearing hosts. METHODS: A matched set of two cloned tumor cell lines (lymphoma+ and lymphoma-), which differ only in MHC Class I gene expression, was established from the parental YAC-1 cell line (a target widely accepted as being sensitive to murine NK cells). An in vivo rapid elimination assay (REA) was performed using tumor cells labeled with [125I] 5-iodo-2 deoxyuridine to evaluate intracerebral NK cell-mediated defense immunity. There was no difference in the in vitro growth rate and c-myc gene expression between lymphoma+ and lymphoma- cells. An in vitro cytotoxicity assay showed that the lymphoma+ cell line was sensitive to MHC Class I-restricted CTL-mediated lysis, whereas the lymphoma- line was refractory to it. Both were susceptible to NK cell mediated lysis, comparable to the level shown by YAC-1 cells. Flow cytometry revealed that lymphoma+ reacted positively for cell-surface MHC Class I molecules, whereas lymphoma- had no reaction. Four- to 72-hour REAs, performed using either cell line, disclosed no clearance of radiolabeled tumor cells from the brain in independent groups of untreated and T cell-depleted mice; this contrasted with eradication of radioactivity from the lungs. In NK cell-depleted mice, however, there was no elimination of radiolabeled tumor cells from the brain or lungs. The MHC Class I expression on lymphoma+ cells was enhanced after intracerebral inoculation, rendering them less sensitive to NK cells. By contrast, lymphoma- cells remained negative for cell-surface MHC expression, being sensitive to NK cells and refractory to CTLs after intracerebralinoculation. These results indicate the absence of NK cell-mediated lytic activity in the brain. This allows even NK cell-sensitive tumor cells to escape intracerebral immunosurveillance. CONCLUSIONS: These experiments have refined the information that the brain may lack NK cell-mediated defense immunity against intracerebrally growing tumors, representing a characteristic aspect of this immunologically privileged organ. PMID- 12650435 TI - Neurogenic motor evoked potentials: role in brachial plexus surgery. Case report. AB - Peripheral nerve graft repair after severe brachial plexus injury is futile if there is degeneration of motor fibers in the proximal nerve stump to which the graft must be attached. Traditional intraoperative neurophysiological assessment methods like nerve action potential (NAP) and somatosensory evoked potential (SSEP) monitoring have been used to evaluate proximal nerve stump integrity, but these methods do not allow evaluation of the integrity of motor fibers back to the anterior horn cell. Consequently, the authors used transcranial electrical stimulation and recorded neurogenic motor evoked potentials (MEPs) directly from the brachial plexus in a patient undergoing surgical repair of a complete upper brachial plexus injury (Erb palsy) to assess the functional continuity of motor fibers. In addition, selected elements of the brachial plexus were directly stimulated, and NAPs were recorded. Finally, SSEPs were recorded from the scalp after stimulation of selected elements of the brachial plexus. Neurogenic MEPs were present from the medial cord of the brachial plexus, but not the middle or upper trunk; NAPs were present from the lateral and posterior cords after middle trunk stimulation, but absent after upper trunk stimulation; and SSEPs were present after medial cord stimulation but absent after stimulation of the upper and middle trunks. For the first time, neurogenic MEPs were coupled with NAPs and SSEPs to evaluate successfully the functional status of motor fibers back to the anterior horn cell for accurate localization of the lesion sites. PMID- 12650437 TI - Selective loss of Purkinje cells in transverse and sigmoid dural arteriovenous fistulas. Report of two cases. AB - The histological changes that occur in brain tissue have rarely been documented in patients with dural arteriovenous fistulas (AVFs). In this study the authors report on two patients with dural AVFs in the transverse-sigmoid sinus who presented with subarachnoid hemorrhage or progressive dementia. Histological studies of the cerebellar cortices showed a selective loss of Purkinje cells, indicating an ischemic insult caused by venous hypertension. Admission N isopropyl-p-123I-iodoamphetamine single-photon emission computerized tomography scans demonstrated a decrease in cerebral blood flow, including flow through the cerebellum. Venous hypertension caused by transverse-sigmoid sinus dural AVFs provokes an ischemic condition severe enough to cause selective neuronal damage in the cerebellum. PMID- 12650436 TI - Recombinant activated factor VII for cerebral injury-induced coagulopathy in pediatric patients. Report of three cases and review of the literature. AB - Brain injury remains one of the leading causes of death and disability in children. Appropriate therapy involves aggressive management of intracranial pressure (ICP) and cerebral perfusion pressure, which often requires placement of an intraparenchymal ICP monitor or intraventricular catheter. These potentially life-saving interventions require normal coagulation function; however, several factors may lead to coagulopathy in the head-injured patient. Standard therapies, which often include multiple doses of fresh frozen plasma (FFP), have a number of drawbacks when used in the pediatric population. The use of FFP requires time to type and crossmatch, thaw, and administer. It imposes a significant volume load on a child in whom cerebral edema remains a problem. Success in using recombinant activated factor VII (rFVIIa) in the hemophiliac population suggests an alternative therapy. Three patients suffered severe coagulopathy after cerebral injury. One patient received rFVIIa after repeated doses of FFP had failed to correct the coagulopathy; the other two patients received rFVIIa as the initial therapy. Treatment with rFVIIa consisted of a bolus of 90 microg/kg. Recombinant activated factor VII rapidly corrected the patients' coagulopathies, which allowed placement of intraparenchymal fiberoptic lines and intraventricular catheters to monitor ICP. The patients suffered no complication from the placement of ICP monitoring devices, as demonstrated on computerized tomography scans obtained within 24 hours after placement. Brain injury-induced coagulopathy may lead to significant secondary injury and delays the invasive monitoring necessary for the aggressive management of intracranial hypertension. Fresh frozen plasma takes time to administer. may require repeated doses of significant volume for the pediatric patient, and may ultimately fail. Preliminary data indicated that rFVIIa provides a rapid and successful correction of coagulopathy in the head-injured patient. PMID- 12650438 TI - Intraparenchymal schwannoma of the medulla oblongata. Case report. AB - A 48-year-old man presented with a rare intraparenchymal schwannoma of the medulla oblongata. After he underwent gamma knife surgery, the patient's condition deteriorated and the mass, which at the time was thought to be a glioma, became larger on magnetic resonance images. The mass was resected through a suboccipital craniectomy via the transcerebellomedullary fissure approach. The tumor, which was moderately firm but distinct from the surrounding parenchyma, was removed totally. Postoperative histological and immunohistochemical examinations confirmed the diagnosis of a benign schwannoma. Brainstem schwannomas can be cured by microneurosurgery. It is important to distinguish these tumors from glioma. The main theories on the cause(s) of this lesion are briefly reviewed. PMID- 12650439 TI - "Real" three-dimensional constructive interference in steady-state imaging to discern microneurosurgical anatomy. Technical note. AB - Three-dimensional (3D) neuroimages are generally considered useful for neurosurgical practice. Nevertheless, neuroimaging modalities such as 3D digital subtraction angiography and 3D computerized tomography angiography are still insufficient because the resulting images fail to delineate neural structures. Complex neurosurgical procedures are mostly performed in the cerebrospinal fluid (CSF) space of the basal cistern, where vessels and neural structures are present along with the lesion. The magnetic resonance (MR) imaging-derived 3D constructive interference in steady-state (CISS) imaging displays the margin between the CSF and neural structures, vessels, and dura mater in detail, in a two-dimensional fashion. The authors know that volume-rendered 3D CISS images would be more useful for surgery than conventional ones. Although the usefulness of "virtual MR image endoscopy" was reported previously, the endoscopic view is different from the operative field because of the perspective being emphasized. Therefore, to simulate surgical views, the authors made 3D neuroimages from a 3D CISS MR sequence by using an advanced computer workstation. After generating volume images, a cutting method was used in the desired plane to visualize the lesion with reference to a multiplanar reformatted image. The authors call these "real" 3D CISS images, and they are more comparable to the operative field. This newly developed method of producing a real 3D CISS image was used in 30 cases and contributed to the understanding of the relationship between a lesion and surrounding structures before attempting neurosurgical procedures, with minimal invasiveness to the patient. PMID- 12650440 TI - Theodore Brown Rasmussen (1910-2002): epilepsy surgeon, scientist, and teacher. AB - Theodore Brown Rasmussen succeeded Wilder Penfield as director of the Montreal Neurological Institute (MNI) and held this post from 1960 to 1972. During his career, Rasmussen probably performed more operations for epilepsy than any other surgeon of his time; he became the foremost authority in this field. His meticulous follow-up analyses of the MNI seizure series provided substantial evidence for the success of surgery in the treatment of focal epilepsy. In addition, he made significant contributions to surgery of the pituitary gland for control of cancer, treatment of cerebral and spinal tumors, application of the intracarotid Amytal test for lateralization of speech and memory function, and characterization and treatment of epilepsy accompanied by chronic encephalitis, now referred to as Rasmussen syndrome. His painstaking attention to surgical details as well as his insistence on close monitoring of patient care and critical scrutiny of clinical results marked him as an outstanding teacher and role model for young neurosurgeons and neuroscientists. PMID- 12650441 TI - Aneurysm clips. AB - This communication outlines the development of aneurysm clips, from those originally used by Walter Dandy to those in use today. The history is rich, with many contributions from neurosurgical pioneers and innovators. As a result, the modern neurosurgeon has a wide selection of biocompatible aneurysm clips from which to choose, clips that have known closing pressures and various sizes and shapes, as well as a selection of clip applicators that do not obstruct the surgical field. PMID- 12650442 TI - Unusual venous drainage of an arteriovenous malformation. Case illustration. PMID- 12650443 TI - Paget disease causing intracerebral mucoceles. Case illustration. PMID- 12650444 TI - Trigeminal neuralgia. PMID- 12650445 TI - Intraventricular neurocysticercosis. PMID- 12650446 TI - Death and third ventriculostomy. PMID- 12650447 TI - Death and third ventriculostomy. PMID- 12650448 TI - Field potentials. PMID- 12650449 TI - Erwinia carotovora subsp. carotovora and Erwinia-derived elicitors HrpN and PehA trigger distinct but interacting defense responses and cell death in Arabidopsis. AB - We have used an hrp-positive strain of the soft rot pathogen Erwinia carotovora subsp. carotovora to elucidate plant responses to this bacterial necrotroph. Purified virulence determinants, harpin (HrpN) and polygalacturonase (PehA), were used as tools to facilitate this analysis. We show that HrpN elicits lesion formation in Arabidopsis and tobacco and triggers systemic resistance in Arabidopsis. Establishment of resistance is accompanied by the expression of salicylic acid (SA)-dependent, but also jasmonate/ethylene (JA/ET)-dependent, marker genes PR1 and PDF1.2, respectively, suggesting that both SA-dependent and JA/ET-dependent defense pathways are activated. Use of pathway-specific mutants and transgenic NahG plants show that both pathways are required for the induction of resistance. Arabidopsis plants treated simultaneously with both elictors PehA, known to trigger only JA/ET-dependent defense signaling, and HrpN react with accelerated and enhanced induction of the marker genes PR1 and PDF1.2 both locally and systemically. This mutual amplification of defense gene expression involves both SA-dependent and JA/ET-dependent defense signaling. The two elicitors produced by E. carotovora subsp. carotovora also cooperate in triggering increased production of superoxide and lesion formation. PMID- 12650450 TI - The 50-kDa protein of Apple chlorotic leaf spot virus interferes with intracellular and intercellular targeting and tubule-inducing activity of the 39 kDa protein of Grapevine berry inner necrosis virus. AB - To understand why transgenic Nicotiana occidentalis plants expressing a functional movement protein (MP) of Apple chlorotic leaf spot virus (ACLSV) show specific resistance to Grapevine berry inner necrosis virus (GINV), the MPs of ACLSV (50KP) and GINV (39KP) were fused to green, yellow, or cyan fluorescent proteins (GFP, YFP, or CFP). These fusion proteins were transiently expressed in leaf cells of both transgenic (50KP) and nontransgenic (NT) plants, and the intracellular and intercellular trafficking and tubule-inducing activity of these proteins were compared. The results indicate that in epidermal cells and protoplasts from 50KP plant leaves, the trafficking and tubule-inducing activities of GINV-39KP were specifically blocked while those of ACLSV-50KP and Apple stem grooving virus MP (36KP) were not affected. Additionally, when 39KP YFP and 50KP-CFP were coexpressed in the leaf epidermis of NT plants, the fluorescence of both proteins was confined to single cells, indicating that 50KP CFP interferes with the cell-to-cell trafficking of 39KP-YFP and vice versa. Mutational analyses of 50KP showed that the deletion mutants that retained the activities described above still blocked cell-to-cell trafficking of 39KP, but the dysfunctional 50KP mutants could no longer impede cell-to-cell movement of 39KP. Transgenic plants expressing the functional 50KP deletion mutants showed specific resistance against GINV. In contrast, transgenic plants expressing the dysfunctional 50KP mutants did not show any resistance to the virus. From these results, we conclude that the specific resistance of 50KP plants to GINV is due to the ability of the 50KP to block intracellular and intercellular trafficking of GINV 39KP. PMID- 12650451 TI - Expression of peroxidase-like genes, H2O2 production, and peroxidase activity during the hypersensitive response to Xanthomonas campestris pv. vesicatoria in Capsicum annuum. AB - Pepper ascorbate peroxidase-like (CAPOA1), thioredoxin peroxidase-like (CAPOT1), and peroxidase-like (CAPO1) clones were isolated from pepper leaves inoculated with avirulent strain Bv5-4a of Xanthomonas campestris pv. vesicatoria. CAPOA1, CAPOT1, and CAPO1 mRNA disappeared 18 to 30 h after the bacterial infection when the hypersensitive response (HR) was visible. In contrast, peroxidase activity reached a peak at 18 h after infection and then declined at 24 and 30 h when H2O2 accumulation level was maximal. These results suggest that the striking accumulation of H2O2 and strong decrease in peroxidase activity during the programmed cell death may be due to the strong suppression of CAPOA1, CAPOT1, and CAPO1 gene expression. Infection by Phytophthora capsici or Colletotricum gloeosporioides also induced the expression of the three putative peroxidase genes in pepper tissues. CAPOA1 mRNAs were in situ localized in phloem areas of vascular bundles in pepper tissues infected by Colletotricum. coccodes, P. capsici, or C. gloeosporioides. Exogenous treatment with H2O2 strongly induced the CAPOA1 and CAPOT1 transcription 1 h after treatment, while the CAPO1 transcripts accumulated 12 h after H2O2 treatment. We suggest that pepper ascorbate peroxidase and thioredoxin peroxidase genes may function as regulators of H2O2 level and total peroxidase activity in the oxidative burst during the HR to incompatible pathogen interaction in pepper plant. PMID- 12650452 TI - Analysis of the involvement of an inducible Arabidopsis RNA-dependent RNA polymerase in antiviral defense. AB - RNA-dependent RNA polymerases (RdRPs) have been implicated in posttranscriptional gene silencing (PTGS) and antiviral defense. An Arabidopsis RdRP (SDE1/SGS2) has been previously shown to be required for transgene-induced PTGS but has no general role in antiviral defense. On the other hand, we have recently shown that transgenic tobacco deficient in an inducible RdRP (NtRdRP1) activity became more susceptible to both Tobacco mosaic virus and Potato virus X. Thus, different RdRPs may have distinct roles in closely related PTGS and antiviral defense. In the present study, we analyzed roles of a newly identified Arabidopsis RdRP gene (AtRdRP1) in plant antiviral defense. AtRdRP1 encodes an RdRP closely related structurally to NtRdRP1 and is also induced by salicylic acid treatment and virus infection. A T-DNA insertion mutant for AtRdRP1 has been isolated and analyzed for possible alterations in response to viral infection. When infected by a tobamovirus and a tobravirus, the knockout mutant accumulated higher and more persistent levels of viral RNAs in both the lower, inoculated and in upper, systemically infected leaves than did wild-type plants. These results suggest that the inducible AtRdRP1 is the Arabidopsis ortholog of NtRdRP1 and plays a role in antiviral defense. Examination of short viral RNAs and silencing studies using a viral vector harboring an endogenous plant gene suggest that, while not required for virus-induced PTGS, AtRdRP1 can apparently promote turnover of viral RNAs in infected plants. PMID- 12650453 TI - Expression of the bacterial catalase genes during Sinorhizobium meliloti-Medicago sativa symbiosis and their crucial role during the infection process. AB - Sinorhizobium meliloti possesses three distinct catalases to cope with oxidative stress: two monofunctional catalases (KatA and KatC) and one bifunctional catalase-peroxydase (KatB). The katB gene is constitutively expressed during growth in batch culture and is not induced under oxidative stress conditions. In contrast, the expression of katA and katC genes is mainly regulated at the transcription level in these conditions. A differential expression of kat genes was observed during the development of the nodule. A high expression of katA gene was detected in bacteroids, suggesting that the nitrogen-fixation process induces a strong oxidative stress. In contrast, bacteria express katB and katC genes and not the H2O2-inducible katA gene in infection threads despite the detection of H2O2 around the bacteria. A katB katC double mutant nodulated poorly and displayed abnormal infection. After nonefficient release into plant cells, bacteria failed to differentiate into bacteroids and rapidly underwent senescence. Our results indicate that these two catalases are essential for the establishment of the symbiosis. They also suggest that the bacteria are in a nonexponential growth phase in infection threads and corroborate previous studies on the growth rate of bacteria inside the plant. PMID- 12650454 TI - Comparative study of regulatory mechanisms for pectinase production by Erwinia carotovora subsp. carotovora and Erwinia chrysanthemi. AB - The production of pectinase, the major virulence determinant of soft-rot Erwinia species, is controlled by many regulatory factors. We focused on the major regulatory proteins, KdgR, CRP, Pir, and PecS, characterized mainly in E. chrysanthemi, and tested for their presence and function in the control of pectate lyase (Pel) and polygalacturonase (Peh) production in E. carotovora subsp. carotovora. Homologues of kdgR and crp but not of pir and pecS were detected by Southern blot analyses in E. carotovora subsp. carotovora. In fact, KdgR and CRP homologues of E. carotovora subsp. carotovora had high amino acid identities to those of E. chrysanthemi, including a complete match of the hypothetical helix-turn-helix DNA-binding motif. However, in Western blot analyses using anti-Pir (E. chrysanthemi) antibodies, a cross-reacting protein was present in both Erwinia species, although Pel production in E. carotovora subsp. carotovora was not further stimulated by adding plant extract into the medium containing PGA (polygalacturonic acid) in which hyperinduction by Pir has been reported in E. chrysanthemi EC16. When plasmids that contained each of these regulatory genes from E. chrysanthemi were introduced into E. carotovora subsp. carotovora, Pel production was controlled as predicted from their roles in E. chrysanthemi, except for PecS. PecS exerted a positive control in E. carotovora subsp. carotovora, in contrast to a negative control in E. chrysanthemi. DNA binding assays demonstrated that KdgR, CRP, Pir, and PecS of E. chrysanthemi and KdgR and CRP homologues of E. carotovora subsp. carotovora could bind to the promoter regions of pel-1, pel-3, and peh of E. carotovora subsp. carotovora. Taken together, KdgR and CRP homologues of E. carotovora subsp. carotovora may regulate Pel and Peh production as in E. chrysanthemi. However, the presence of Pir and PecS homologues in E. carotovora subsp. carotovora was not identified in this study, though these proteins of E. chrysanthemi were functional on the promoter regions of the pectinase genes of E. carotovora subsp. carotovora. PMID- 12650455 TI - The HrpX/HrpY two-component system activates hrpS expression, the first step in the regulatory cascade controlling the Hrp regulon in Pantoea stewartii subsp. stewartii. AB - A regulatory cascade activating hrp/hrc type III secretion and effector genes was delineated in Pantoea stewartii subsp. stewartii, a bacterial pathogen of corn. Four hrp regulatory genes were characterized: hrpX and hrpY encode the sensor kinase and response regulator, respectively, of a two-component signal transduction system; hrpS encodes an NtrC-like transcriptional enhancer; and hrpL encodes an alternative sigma factor. Epistasis analysis, expression studies using gene fusions, and genetic reconstruction of each step in Escherichia coli were used to delineate the following pathway: HrpY activates hrpS and also positively autoregulates the hrpXY operon. In turn, HrpS is required for full activation of the sigma54-dependent hrpL promoter. Finally, HrpL controls expression of all known hrp and wts genes. In vitro, hrpS and all downstream hrp genes were regulated by pH and salt concentration. Mutants with in-frame deletions in hrpX were still partially virulent on corn but were unable to sense the chemical or metabolic signals that induce hrp genes in vitro. Site-directed mutagenesis of HrpY indicated that aspartate 57 is the probable phosphorylation site and that it is needed for activity. These findings suggest that both HrpX and an alternate mechanism are involved in the activation of HrpY in planta. PMID- 12650457 TI - The 2b protein of cucumoviruses has a role in promoting the cell-to-cell movement of pseudorecombinant viruses. AB - Pseudorecombinant viruses (i.e., those containing a reassorted genome of closely related multipartite viruses) are often not as competitive as the parental viruses. The role of the 2b gene in hypervirulence and maintenance of a progressive infection was assessed in a pseudorecombinant virus formed between RNAs 1 plus 2 of Cucumber mosaic virus (CMV) and RNA 3 of Tomato aspermy virus (TAV). The presence of RNA 3 of TAV was found to affect the level of RNA accumulation but not the level of virulence. By contrast, the 2b genes of both TAV and a hypervirulent strain of CMV (WAII-CMV) were found to affect the virulence of the pseudorecombinant viruses but not the levels of viral RNA accumulation. The 2b gene rather than the overlapping open reading frame encoding the C-terminal 41 amino acids of 2a protein of the corresponding virus was found to be essential for promoting infection of the pseudorecombinant viruses in planta. However, the 2b gene was not essential for replication of pseudorecombinant viruses containing CMV RNAs 1 plus 2 and TAV RNA 3. These results indicate that the 2b protein is involved in promoting the cell-to-cell movement of the pseudorecombinant viruses. These data also suggest the existence of specific interaction between the TAV 2b protein and either RNA 3 or its encoded proteins, which may be critical for promoting or maintaining infection or both. PMID- 12650456 TI - The regulatory cascade that activates the Hrp regulon in Erwinia herbicola pv. gypsophilae. AB - The pathogenicity of Erwinia herbicola pv. gypsophilae (Ehg) is dependent on a plasmid (pPATH(Ehg)) that harbors the hrp gene cluster and additional virulence genes. The hrp regulatory cascade of Ehg comprises an hrpXY operon encoding a two component system; hrpS encoding a transcriptional factor of the NtrC family and hrpL encoding an alternative sigma factor. Results obtained suggest the following signal transduction model for activating the Hrp regulon: phosphorylated HrpY activates hrpS, HrpS activates hrpL, and HrpL activates genes containing "hrp box" promoter. This model was supported by studies on the effects of mutations in the regulatory genes on pathogenicity and complementation analysis. Nonpolar mutations in hrpX did not affect virulence or transcription of downstream genes. Site-directed mutagenesis of the conserved aspartate 57 in HrpY suggested that its phosphorylation is crucial for activating the hrp regulatory cascade. Studies on the effects of mutations in the hrp regulatory genes on transcriptional activity of downstream genes or of their isolated promoters in planta showed dependency of hrpS expression on active HrpY, of hrpL expression on active HrpS, and of hrpN or hrpJ expression on active HrpL. These results were also partially supported by overexpression of regulatory genes under in vitro conditions. The hrpXY is constitutively expressed with high basal levels under repressive conditions, in contrast to hrpS and hrpL, which exhibit low basal expression levels and are environmentally regulated. PMID- 12650458 TI - 2002 Sewall Wright Award. Linda Partridge. PMID- 12650459 TI - The ecology of individuals: incidence and implications of individual specialization. AB - Most empirical and theoretical studies of resource use and population dynamics treat conspecific individuals as ecologically equivalent. This simplification is only justified if interindividual niche variation is rare, weak, or has a trivial effect on ecological processes. This article reviews the incidence, degree, causes, and implications of individual-level niche variation to challenge these simplifications. Evidence for individual specialization is available for 93 species distributed across a broad range of taxonomic groups. Although few studies have quantified the degree to which individuals are specialized relative to their population, between-individual variation can sometimes comprise the majority of the population's niche width. The degree of individual specialization varies widely among species and among populations, reflecting a diverse array of physiological, behavioral, and ecological mechanisms that can generate intrapopulation variation. Finally, individual specialization has potentially important ecological, evolutionary, and conservation implications. Theory suggests that niche variation facilitates frequency-dependent interactions that can profoundly affect the population's stability, the amount of intraspecific competition, fitness-function shapes, and the population's capacity to diversify and speciate rapidly. Our collection of case studies suggests that individual specialization is a widespread but underappreciated phenomenon that poses many important but unanswered questions. PMID- 12650460 TI - Unequal transmission of mitochondrial haplotypes in natural populations of field mice with XY females (genus Akodon). AB - In species with fertile XY females, such as South American field mice (genus Akodon), there are two types of mitochondrial DNA (mtDNA), one passing from XX females and one from XY females. The XX mothers pass their mtDNA to their XX daughters. The XY mothers, however, produce both XX and XY daughters. Because of this breeding scheme, the XY mtDNA remains isolated whereas the XX lineage is continuously invaded by XY mtDNA haplotypes. Using a set of recursion equations, I predicted that XY mtDNA haplotypes should rapidly spread through entire populations composed of both XX and XY females. I examined patterns of nucleotide polymorphism and divergence from the mtDNA control region as well as phylogenetic patterns for evidence of an mtDNA sweep. I compared patterns in two sister species, Akodon boliviensis and Akodon azarae, that are composed of 35% and 10% XY females, respectively. Akodon boliviensis XY females are found in all clades of a phylogenetic mtDNA tree consistent with the spread of mtDNA haplotypes. In addition, A. azarae mtDNA haplotypes showed no deviations from neutrality. These results, in combination with high levels of mtDNA nucleotide diversity in XY females, suggest an ancient origin (>10(4) generations) of XY females in both A. boliviensis and A. azarae. PMID- 12650461 TI - Does herbivore diversity depend on plant diversity? The case of California butterflies. AB - It is widely believed that the diversity of plants influences the diversity of animals, and this should be particularly true of herbivores. We examine this supposition at a moderate spatial extent by comparing the richness patterns of the 217 butterfly species resident in California to those of plants, including all 5,902 vascular plant species and the 552 species known to be fed on by caterpillars. We also examine the relationships between plant/butterfly richness and 20 environmental variables. We found that although plant and butterfly diversities are positively correlated, multiple regression, path models, and spatial analysis indicate that once primary productivity (estimated by a water energy variable, actual evapotranspiration) and topographical variability are incorporated into models, neither measure of plant richness has any relationship with butterfly richness. To examine whether butterflies with the most specialized diets follow the pattern found across all butterflies, we repeated the analyses for 37 species of strict monophages and their food plants and found that plant and butterfly richness were similarly weakly associated after incorporating the environmental variables. We condude that plant diversity does not directly influence butterfly diversity but that both are probably responding to similar environmental factors. PMID- 12650462 TI - Natural selection in a bottle. AB - The study of natural selection in laboratory systems undergoing experimental evolution can provide important insights into the relationship between natural selection and adaptation. We studied selection on the norm of reaction of age at first reproduction in a laboratory population of Drosophila melanogaster. This population had been selected on a discrete generation schedule in the laboratory for more than 600 generations. Using genetically marked strains, we studied development time, size, female fecundity, and viability of flies that began development at different times relative to the initiation of each bottle. Only flies that began development within 30 h of the initiation of the bottle were reliably able to eclose before the next transfer. Theory predicts that flies initiating development around this critical time should decrease size at maturity to ensure eclosion by the 14-d deadline, but late flies are not smaller. This result suggests an unknown constraint on response to selection on age at maturity in this population. Ultimately, laboratory systems provide the best opportunity for the study of natural selection, genetic variation, and evolutionary response in the same population. PMID- 12650463 TI - Comparative studies of evolutionary responses to light environments in Arabidopsis. AB - In this article, we compare the reaction norms to foliage shade (changes in light quality, spatially fine-grained environmental variation) and photoperiod (day length, spatially coarse-grained environmental variation) in several haplotypes of Arabidopsis thaliana from Scandinavia. We found that both across-environment means and phenotypic plasticities evolved continuously and very rapidly within this group. Both character means and trait plasticities were highly integrated, in part as predicted by the adaptive plasticity hypothesis for response to foliage shade (the so-called shade-avoidance syndrome). We found that a significant but small fraction of the variance in across-treatment trait means and plasticities in response to one environmental factor is explained by variation of the same traits in response to the other factor. Genetic relatedness based on chloroplast DNA sequence variation among haplotypes was not associated with variation in across-treatment character means or their plasticities, suggesting that evolution of these characters has occurred on a local geographic scale via reticulation (outcrossing) among maternal lines rather than by the differential survival of selfing lineages. PMID- 12650464 TI - The effect of energy and seasonality on avian species richness and community composition. AB - We analyzed geographic patterns of richness in both the breeding and winter season in relation to a remotely sensed index of seasonal production (normalized difference vegetation index [NDVI]) and to measures of habitat heterogeneity at four different spatial resolutions. The relationship between avian richness and NDVI was consistent between seasons, suggesting that the way in which available energy is converted to bird species is similar at these ecologically distinct times of year. The number and proportion of migrant species in breeding communities also increased predictably with the degree of seasonality. The NDVI was a much better predictor of seasonal richness at finer spatial scales, whereas habitat heterogeneity best predicted richness at coarser spatial resolutions. While we find strong support for a positive relationship between available energy and species richness, seasonal NDVI explained at most 61% of the variation in richness. Seasonal NDVI and habitat heterogeneity together explain up to 69% of the variation in richness. PMID- 12650465 TI - Least-cost input mixtures of water and nitrogen for photosynthesis. AB - In microeconomics, a standard framework is used for determining the optimal input mix for a two-input production process. Here we adapt this framework for understanding the way plants use water and nitrogen (N) in photosynthesis. The least-cost input mixture for generating a given output depends on the relative cost of procuring and using nitrogen versus water. This way of considering the issue integrates concepts such as water-use efficiency and photosynthetic nitrogen-use efficiency into the more inclusive objective of optimizing the input mix for a given situation. We explore the implications of deploying alternative combinations of leaf nitrogen concentration and stomatal conductance to water, focusing on comparing hypothetical species occurring in low- versus high-humidity habitats. We then present data from sites in both the United States and Australia and show that low-rainfall species operate with substantially higher leaf N concentration per unit leaf area. The extra protein reflected in higher leaf N concentration is associated with a greater drawdown of internal CO2, such that low-rainfall species achieve higher photosynthetic rates at a given stomatal conductance. This restraint of transpirational water use apparently counterbalances the multiple costs of deploying high-nitrogen leaves. PMID- 12650466 TI - Explaining species richness from continents to communities: the time-for speciation effect in emydid turtles. AB - Speciation is the process that ultimately generates species richness. However, the time required for speciation to build up diversity in a region is rarely considered as an explanation for patterns of species richness. We explored this "time-for-speciation effect" on patterns of species richness in emydid turtles. Emydids show a striking pattern of high species richness in eastern North America (especially the southeast) and low diversity in other regions. At the continental scale, species richness is positively correlated with the amount of time emydids have been present and speciating in each region, with eastern North America being the ancestral region. Within eastern North America, higher regional species richness in the southeast is associated with smaller geographic range sizes and not greater local species richness in southern communities. We suggest that these patterns of geographic range size variation and local and regional species richness in eastern North America are caused by glaciation, allopatric speciation, and the time-for-speciation effect. We propose that allopatric speciation can simultaneously decrease geographic range size and increase regional diversity without increasing local diversity and that geographic range size can determine the relationship between alpha, beta, and gamma diversity. The time-for-speciation effect may act through a variety of processes at different spatial scales to determine diverse patterns of species richness. PMID- 12650467 TI - Evolution of egg dumping in a subsocial insect. AB - Egg dumping, or abandonment of eggs and young to the care of other conspecifics, frees individuals from costs of maternal care while potentially imposing energetic and ecological costs on egg recipients. It is not clear, however, that egg dumping necessarily represents selfish manipulation of egg recipients, and in some ecological contexts, recipients may benefit from enlarged broods. Thus, egg dumping may either be mutually beneficial for dumpers and recipients or entail costs for dumpers that are compensated by other means, such as improving reproduction of genetically related egg recipients. Here I use field experiments to test the relative importance of manipulation (i.e., "parasitism"), mutualism, and kin selection in the evolution of egg dumping in the tingid lace bug Gargaphia solani. In support of mutualism and kin selection, I found that reproduction of egg recipient G. solani benefits from brood enlargement, most likely because eggs and gregarious nymphs find safety in greater numbers. But contrary to both parasitism and mutualism, egg dumper reproduction was not improved by offspring abandonment. Indeed, dumpers laid smaller clutches than recipients, and dumpers did not convert a survival advantage into greater future reproduction. Genetic analyses of a natural G. solani population revealed, however, that dumpers are related to their egg recipients. Moreover, Hamilton's rule showed that egg-dumping G. solani earn sufficient indirect genetic benefits for kin selection to favor the behavior. Thus, egg dumping in some species may be kin-selected cooperation rather than parasitism or mutualism. PMID- 12650468 TI - Socially stable territories: the negotiation of space by interacting foragers. AB - This article presents a theory of territoriality that integrates optimal foraging and conflict resolution through negotiation. Using a spatially explicit model of a sit-and-wait forager, we show that when resources are scarce, there is a conflict between foragers: there is not enough space for all individuals to have optimal home ranges. We propose that a division of space that solves this conflict over resources is the outcome of a negotiation between foragers. We name this outcome the socially stable territories (SST). Using game theory we show that in a homogenous patch occupied by two interacting foragers, both individuals receive identical energy yields at the socially stable territories; that is, there is economic equity. Economic inequity can arise in a heterogeneous patch or from asymmetries in fighting abilities between the foragers. Opportunity costs play a role in reducing economic inequity. When the asymmetry in fighting abilities is very large, a negotiated division of space is not possible and the forager with lowest fighting ability may be evicted from the habitat patch. A comparison between territories and overlapping home ranges shows that energy yields from territories are generally higher. We discuss why there are instances in which individuals nevertheless overlap home ranges. PMID- 12650469 TI - Consequences of hierarchical allocation for the evolution of life-history traits. AB - Resource allocation within individuals may often be hierarchical, and this may have important effects on genetic correlations and on trait evolution. For example, organisms may divide energy between reproduction and somatic growth and then subdivide reproductive resources. Genetic variation in allocation to pathways early in such hierarchies (e.g., reproduction) can cause positive genetic correlations between traits that trade off (e.g., offspring size and number) because some individuals invest more resources in reproduction than others. We used quantitative-genetic models to explore the evolutionary implications of allocation hierarchies. Our results showed that when variation in allocation early in the hierarchy exceeds subsequent variation in allocation, genetic covariances and initial responses to selection do not reflect trade-offs occurring at later levels in the hierarchy. This general pattern was evident for many starting allocations and optima and for whether traits contributed multiplicatively or additively to fitness. Finally, artificial selection on a single trait revealed masked trade-offs when variation in early allocation was comparable to subsequent variation in allocation. This result confirms artificial selection as a powerful, but not foolproof, method of detecting trade-offs. Thus, allocation hierarchies can profoundly affect life-history evolution by causing traits to evolve in the opposite direction to that predicted by trade-offs. PMID- 12650470 TI - Observations on Salmonella contamination of commercial laying farms before and after cleaning and disinfection. AB - Little is known about the effectiveness of the cleaning and disinfection methods in use on commercial laying farms in Great Britain. Samples were taken from poultry house structures and equipment of five cage layer flocks, five barn egg production flocks and two free-range flocks. In the free-range houses there was a decrease in Salmonella after cleaning and disinfection, although the soil in the paddocks remained contaminated. In the barn and especially the cage layer houses, significant residual contamination remained on the surfaces of buildings and equipment. Wildlife pests were also found to be carrying Salmonella in the disinfected houses and free-range paddocks. PMID- 12650471 TI - Measurement of tidal breathing flow-volume loop indices in horses used for different sporting purposes with and without recurrent airway obstruction. AB - Tidal breathing flow-volume loop (TBFVL) indices in healthy horses and in horses suffering from mild and moderate to severe recurrent airway obstruction (RAO) were measured. In addition, the ability of these indices to differentiate healthy controls from horses with mild RAO by separating the horses according to their usage was investigated. The horses were assigned to three sporting categories: 1 Leisure horses; 2 Horses competing at events with a duration of more than five minutes (eventing, coaching and endurance); and 3 Horses competing at events with a duration of less than five minutes (dressage and show jumping). Flow-volume curves were recorded with an ultrasonic spirometer and TBFVL indices were calculated with specially dedicated software. The sporting usage of the horses had a significant effect on the differentiating ability of TBFVL indices in diagnosing different degrees of RAO. The index TIF50 differentiated control horses from horses with minor RAO in all sporting categories. PMID- 12650472 TI - Early echocardiographic predictors of myxomatous mitral valve disease in dachshunds. AB - Myxomatous mitral valve disease in dogs is heritable, and it is therefore important to detect the early signs of the disease. This study was conducted to assess the predictive value of early echocardiographic and auscultatory signs of mitral valve prolapse, measured in terms of the leaflet thickness, the area of the regurgitant jet, and the intensity of the murmur, on the increases in left ventricular end diastolic diameter (LVEDD) and left atrial diameter (LAD) in a population of 190 clinically healthy dachshunds followed up for three years. The most significant predictor of an increase in LVEDD was the interaction between the index of mitral valve prolapse and the area of the regurgitant jet (P < 0.0001). In dogs with a jet area greater than 50 per cent of the left atrium, the disease progressed more quickly in terms of increases in LVEDD in relation to the severity of the prolapse at the initial examination. In dogs with smaller jets, the initial prolapse index was not significantly associated with increases in LVEDD. The initial index of mitral valve prolapse, the area of the jet and the intensity of the heart murmur were all significant predictors of an increase in LAD. PMID- 12650473 TI - Searching for BSE in sheep: interpreting the results so far. PMID- 12650474 TI - Strain differences in response to propofol, ketamine and medetomidine in rabbits. PMID- 12650475 TI - Uterine rupture in a cat. PMID- 12650476 TI - Chronic arthritis and systemic amyloidosis in three goat kids associated with seroconversion to Erysipelothrix rhusiopathiae. PMID- 12650477 TI - Self-destructive behaviour in ruminants. PMID- 12650478 TI - Kidney transplants in cats. PMID- 12650479 TI - Kidney transplants in cats. PMID- 12650480 TI - Framework for veterinary education and training. PMID- 12650482 TI - RCVS Council elections. PMID- 12650481 TI - Framework for veterinary education and training. PMID- 12650483 TI - Colonisation site of E coli O157 in cattle. PMID- 12650484 TI - Mastitis and somatic cell counts. PMID- 12650485 TI - Raisin poisoning in a dog. PMID- 12650486 TI - Study of localised dermatosis in rabbits caused by myxomatosis. PMID- 12650487 TI - Anaesthetic strategies to reduce perioperative blood loss in paediatric surgery. AB - In adults, a number of measures to reduce perioperative blood loss have been established. These techniques serve to reduce patients' exposure to homologous blood. Most adults are concerned with this issue especially since many patients became infected with human immunodeficiency virus (HIV) during the 1980s through exposure to blood components. While blood-saving strategies are widely used in adults, they are mostly neglected in infants. However, it is these young patients with their whole life in front of them who, it could be argued, would benefit especially from any potentially avoidable infection (HIV, hepatitis, etc.) or immunological complications. In infants and small children, these blood-sparing techniques may not be as effective as in adults and technical limitations may prevent their application. However, some of these measures can be used and may serve to prevent or reduce exposure to homologous blood. In the following review, blood-saving techniques established in adults are described and their applicability for paediatric patients discussed. PMID- 12650488 TI - Pharmacokinetics and pharmacodynamics of the new propofol prodrug GPI 15715 in rats. AB - BACKGROUND AND OBJECTIVE: We studied the pharmacokinetics and pharmacodynamics of GPI 15715 (Aquavan injection), a new water-soluble prodrug metabolized to propofol by hydrolysis. METHODS: Nine adult male Sprague-Dawley rats (398 +/- 31 g) received a bolus dose of 40 mg GPI 15715. The plasma concentrations of GPI 15715 and propofol were determined from arterial blood samples, and the pharmacokinetics of both compounds were investigated using compartment models whereby the elimination from the central compartment of GPI 15715 was used as drug input for the central compartment of propofol. Pharmacodynamics were assessed using the median frequency of the EEG power spectrum. RESULTS: A maximum propofol concentration of 7.1 +/- 1.7 microg mL(-1) was reached 3.7 +/- 0.2 min after bolus administration. Pharmacokinetics were best described by two compartment models. GPI 15715 showed a short half-life (2.9 +/- 0.2 and 23.9 +/- 9.9 min), an elimination rate constant of 0.18 +/- 0.01 min(-1) and a central volume of distribution of 0.25 +/- 0.02 L kg(-1). For propofol, the half-life was 1.9 +/- 0.1 and 45 +/- 7 min, the elimination rate constant was 0.15 +/- 0.02 min(-1) and the central volume of distribution was 2.3 +/- 0.6 L kg(-1). The maximum effect on the electroencephalogram (EEG)--EEG suppression for >4 s- occurred 6.5 +/- 1.2 min after bolus administration and baseline values of the EEG median frequency were regained 30 min later. The EEG effect could be described by a sigmoid Emax model including an effect compartment (E0 = 16.9 +/- 7.9 Hz, EC50 = 2.6 +/- 0.8 microg mL(-1), ke0 = 0.35 +/- 0.04 min(-1)). CONCLUSIONS: Compared with known propofol formulations, propofol from GPI 15715 showed a longer half-life, an increased volume of distribution, a delayed onset, a sustained duration of action and a greater potency with respect to concentration. PMID- 12650489 TI - Effects of propofol on the systolic and diastolic performance of the postischaemic, reperfused myocardium in rabbits. AB - BACKGROUND AND OBJECTIVE: The effect of propofol on myocardial dysfunction during ischaemia and reperfusion is controversial yet important because of its frequent use in cardiac anaesthesia. Although animal studies suggest a free radical scavenging potential, the cardioprotective properties of propofol have not been demonstrated consistently in vivo. Previous studies focused on systolic function while diastolic function may be a more sensitive marker of ischaemic injury. The main aim was to document the effect of propofol on diastolic function in isolated, blood perfused rabbit hearts subjected to moderate global ischaemia and reperfusion. METHODS: Propofol 168 micromol L(-1), or the equivalent of its vehicle, Intralipid, was administered to 34 paced parabiotic Langendorff blood perfused isolated rabbit hearts before and after 30 min of global normothermic ischaemia. Recovery of systolic function was quantified with the maximum rate of rise of left ventricular pressure. Diastolic performance was assessed using the time constant of the decline in left ventricular pressure (tau) and chamber stiffness (VdP/dV at 12 mmHg). RESULTS: Recovery of systolic function during reperfusion was comparable in the two groups. There was no difference in left ventricular pressure between the two groups at any time during the experiments. Chamber stiffness increased significantly during ischaemia and reperfusion in the control group (from 34 +/- 9 to 54 +/- 8 mmHg during ischaemia, and 43 +/- 5 mmHg after 30 min reperfusion; mean +/-95% confidence interval) but not in the propofol-treated group (29 +/- 5, 36 +/- 8 and 30 +/- 8 at baseline, ischaemia and 30 min reperfusion, respectively). CONCLUSIONS: Propofol has no protective effect on active relaxation or on systolic function in the present model, but it reduces ischaemic and postischaemic chamber stiffness. PMID- 12650490 TI - Effects of bupivacaine used with sevoflurane on the rhythm and contractility in the isolated rat heart. AB - BACKGROUND AND OBJECTIVE: The effects of sevoflurane on bupivacaine cardiotoxicity are mainly attributed to systemic effects. The purpose of this study was to investigate the direct myocardial effects of sevoflurane on bupivacaine toxicity. METHODS: Hearts of 30 Wistar albino rats were isolated and mounted on a Langendorff apparatus perfused by modified Tyrode solution. Experimental groups were: a sevoflurane group (Group S, n = 10)--following baseline and 20 min (Stage 1) recordings, sevoflurane was added in doses of 1.4% (1 MAC) and 2.8% (2 MAC). In the two bupivacaine groups, bupivacaine 5 micromol (Group B5, n = 10) and bupivacaine 10 micromol (Group B10, n = 10) was added to the solution at Stage 1, and sevoflurane was added to the system as in Group S. Haemodynamic variables, i.e. heart rate, PR interval, QRS duration, left ventricular systolic pressure, contractility (+dp/dtmax), relaxation, time to reach peak systolic pressure, change in left ventricular diastolic pressure from baseline, and rate-pressure product were recorded. RESULTS: In Group S, there was no change in cardiac rhythm. In bupivacaine groups, severe rhythm disturbances occurred and both the PR intervals and QRS complexes were prolonged significantly. All contractility variables deteriorated and the rate-pressure product decreased by 67-90% with the addition of bupivacaine. In all groups, 2 MAC sevoflurane lowered +dp/dtmax further. CONCLUSIONS: Sevoflurane does not have any untoward effect on bupivacaine-induced cardiotoxicity in clinically relevant doses in the isolated rat heart. PMID- 12650491 TI - Effects of rolipram, pimobendan and zaprinast on ischaemia-induced dysrhythmias and on ventricular cyclic nucleotide content in the anaesthetized rat. AB - BACKGROUND AND OBJECTIVE: This study was designed to compare the haemodynamic, electrophysiological and pharmacodynamic effects of three selective inhibitors of the different isoenzyme forms of phosphodiesterase (PDE) on ischaemia-induced dysrhythmias in the anaesthetized rat. The drugs used were pimobendan, a selective PDE III inhibitor, rolipram, a selective PDE IV inhibitor, and zaprinast, a selective PDE V inhibitor. METHODS: The coronary artery was occluded 15 min after commencing drug administration, and myocardial ischaemia was maintained for 30 min during which the heart rate and mean arterial pressure were recorded. cAMP and cGMP were determined by radioimmunoassay. RESULTS: Pretreatment with rolipram decreased the duration of ventricular tachycardia without any change in the incidences of dysrhythmias or the mortality rate. This drug did not modify ventricular content of adenosine 3',5'-cyclic monophosphate (cAMP) or guanosine 3',5'-cyclic monophosphate (cGMP). Pimobendan (1 mg kg(-1) + 0.1 mg kg(-1) min) decreased the duration of ventricular tachycardia. This dose of pimobendan and zaprinast (1 mg kg(-1) + 0.1 mg kg(-1) min(-1)) increased the incidence rate of ventricular fibrillation following coronary artery ligation and the mortality rate. Moreover, both drugs increased cGMP in the ventricle. CONCLUSIONS: The results demonstrated that pimobendan and zaprinast increased the incidence of dysrhythmias and the mortality rate, which was accompanied by an increase in the ventricular content of cGMP. Rolipram decreased the duration of ventricular tachycardia without a change in the cyclic nucleotide content or in the mortality rate. PMID- 12650492 TI - Effect of the neuronal nitric oxide synthase inhibitor 7-nitroindazole on the righting reflex ED50 and minimum alveolar concentration during sevoflurane anaesthesia in rats. AB - BACKGROUND AND OBJECTIVE: The aim was to determine the effect of acute and chronic administration of 7-nitroindazole, a selective neuronal nitric oxide synthase inhibitor, on the righting reflex ED50 and the minimum alveolar concentration during sevoflurane anaesthesia in rats. METHODS: 7-Nitroindazole was acutely (0, 50 and 100 mg kg(-1)) and chronically (0 and 150 mg kg(-1) day( 1), 4 days) administered to rats. After the preparation, the minimum alveolar concentration and the righting reflex ED50 were measured. The concentration of cGMP in the brain, cerebellum and spinal cord was also measured. RESULTS: Acute administration reduced the minimum alveolar concentration (50 mg kg(-1), 58.8% (95% CI: 50.3-67.3%) of the baseline value, P < 0.01; 100 mg kg(-1), 55.8 (46.9 64.7), P < 0.01) and the righting reflex ED50 (50 mg kg(-1), 27.2 (17.2-37.2), P < 0.01; 100 mg kg(-1), 14.3 (6.6-22.0), P < 0.01). Chronic administration did not reduce the minimum alveolar concentration; however, it reduced the righting reflex ED50 (65.3 (52.9-77.7), P < 0.01). Overall, the reduction in minimum alveolar concentration in the acute and chronic protocol did not correlate with that of the righting reflex ED50. 7-Nitroindazole (100 mg kg(-1), acute) reduced the cGMP concentration within the cerebellum by 55.4%; however, it did not decrease concentrations in the brain or spinal cord. CONCLUSIONS: Different mechanisms are responsible for the observed alterations to the minimum alveolar concentration and the righting reflex ED50 following treatment with 7 nitroindazole. The nitric oxide-cGMP pathway might play a less important role in the determination of minimum alveolar concentration than the righting reflex ED50. PMID- 12650493 TI - High-affinity block of voltage-operated rat IIA neuronal sodium channels by 2,6 di-tert-butylphenol, a propofol analogue. AB - BACKGROUND AND OBJECTIVE: Propofol is a phenol derivative (2,6 di isopropylphenol) with a unique effect profile including activating effects on GABA(A) and blocking effects on voltage-operated sodium channels. If the substituents in the 2- and the 6-positions are replaced by tert-butyl groups, the resulting phenol derivative, 2,6 di-tert-butylphenol, despite being a close structural propofol analogue, completely lacks GABA(A) receptor effects. The aim of this in vitro study was to investigate the effects of 2,6 di-tert-butylphenol on voltage-operated neuronal sodium channels in order to determine whether and, if so, how these structural changes alter the sodium channel-blocking effect seen with propofol. METHODS: Whole-cell sodium inward currents through heterologously expressed rat type IIA sodium channels were recorded in the absence and presence of definite concentrations of 2,6 di-tert-butylphenol and propofol. RESULTS: When applied at concentrations > or = 30 micromol, 2,6 di-tert-butylphenol completely and irreversibly blocked sodium inward currents. The blockade equilibrium time was about 2 min. A partial washout was possible only if the application was stopped before the equilibrium of the blockade was achieved. CONCLUSIONS: 2,6 Di tert-butylphenol exerts a high-affinity block of neuronal sodium channels. Apparently, the slight structural differences of 2,6 di-tert-butylphenol in comparison with propofol--which account for the lack of GABA(A) receptor effects- enhance its voltage-operated sodium channel-blocking effects. As 2,6 di-tert butylphenol is much more potent than most sodium channel blockers in clinical use, it might be of interest in the development of local anaesthetics. PMID- 12650494 TI - Factors determining the duration of tracheal intubation in cardiac surgery: a single-centre sequential patient audit. AB - BACKGROUND AND OBJECTIVE: The study was designed to identify those factors associated with early tracheal extubation following cardiac surgery. Previous studies have tended to concentrate on surgery for coronary artery bypass or on other selected cohorts. METHODS: Sequential cohort analysis of 296 unselected adult cardiac surgery patients was performed over 3 months. RESULTS: In total, 39% of all patients were extubated within 6 h, 89% within 24 h and 95% within 48 h. Delayed extubation (>6 h after surgery) appeared unrelated to age, gender, body mass index, a previous pattern of angina or myocardial infarction, diabetes, preoperative atrial fibrillation, and preoperative cardiovascular assessment, as well as other factors. Delayed tracheal extubation was associated with poor left ventricular, renal and pulmonary function, a high Euroscore, as well as the type, duration and urgency of surgery. Early extubation (<6 h) was not associated with a reduced length of stay in either the intensive care unit or in hospital compared with patients who were extubated between 6 and 24 h. In these groups, it is presumed that organizational and not clinical factors appear to be responsible for a delay in discharge from intensive care. Patients who were extubated after 24 h had a longer duration of hospital stay and a greater incidence of postoperative complications. Postoperative complications were not adversely affected by early tracheal extubation. CONCLUSIONS: In an unselected sequential cohort, both patient- and surgery-specific factors may be influential in determining the duration of postoperative ventilation of the lungs following cardiac surgery. In view of the changing nature of the surgical population, regular re-evaluation is useful in reassessing performance. PMID- 12650495 TI - Suprascapular nerve block or a piroxicam patch for shoulder tip pain after day case laparoscopic surgery. AB - BACKGROUND AND OBJECTIVE: The reported incidence of shoulder tip pain following laparoscopic surgery varies from 35 to 63%. This study evaluated the analgesic efficacy of either performing a prophylactic suprascapular nerve block with bupivacaine or applying a piroxicam patch to the skin over both shoulders for the relief of shoulder tip pain after laparoscopy. METHODS: Sixty healthy informed female patients were randomly assigned to one of three groups: (a) a control group (n = 20), no treatment; (b) a suprascapular nerve block group (n = 20) in which a bilateral suprascapular nerve block was performed before induction of anaesthesia with 5 mL 0.5% bupivacaine with epinephrine; and (c) a piroxicam patch group (n = 20) in which a 48 mg piroxicam patch on the skin of each shoulder was applied before induction of anaesthesia. All patients received a total intravenous anaesthesia technique with propofol, fentanyl and vecuronium. Shoulder tip and wound pain were recorded on a visual analogue pain scale at five time intervals for 24 h after surgery. RESULTS: A total of 80% of patients in the control group, 75% in the suprascapular nerve block group and 45% in the piroxicam patch group complained of shoulder tip pain during the recording period (P < 0.05). The scores for shoulder tip pain in the piroxicam patch group were significantly lower compared with the control group at 3, 6 and 12 h, and compared with the suprascapular nerve block group at 6 and 12 h. The need for analgesics was also significantly lower in the piroxicam patch group compared with the other two groups. CONCLUSIONS: Prophylactic piroxicam patches are effective and safe for the relief of shoulder tip pain after laparoscopy. Bilateral suprascapular nerve block is not effective in this setting. PMID- 12650496 TI - Treatment of hiccup during general anaesthesia or sedation: a qualitative systematic review. AB - BACKGROUND AND OBJECTIVE: Acute hiccup is a minor complication that can occur during sedation or general anaesthesia. The disorder can disturb the surgical field, might interfere with lung ventilation or could hamper diagnostic procedures. The objective was to perform a systematic search for interventions aimed at treating hiccup occurring during anaesthesia or sedation. METHODS: A systematic search for reports describing interventions to treat hiccup in conjunction with anaesthesia was carried out (MEDLINE, EMBASE, Cochrane-Library, manual screening of reference lists and review articles, up to December 2001). Search terms were 'hiccup', 'singultus' or 'hiccough'. RESULTS: Twenty-six reports involving approximately 581 patients focused on hiccup remedies in the anaesthesia setting. Only one report was substantiated by a randomized controlled trial. This investigated methylphenidate 10 mg intravenously in 51 patients, which did not show a beneficial effect compared with placebo. Hiccup was a self limiting phenomenon. Case series and case reports focused on various systemically applied drugs in 12 reports, stimulating techniques (e.g. pharyngeal stimulation) in seven, topical applied remedies (e.g. intranasal ice-cold water) in four, and ventilation techniques (e.g. continuous positive pressure ventilation) in two. CONCLUSIONS: A large variety of interventions have been proposed for the treatment of hiccup during anaesthesia and sedation. However, perioperative treatment is still based on empirical findings and no treatment is 'evidence based'. Thus, no valid recommendations for the treatment of hiccup can be derived. Uncontrolled observations are inadequate to establish treatment efficacy. PMID- 12650498 TI - Central neuraxial blocks and anticoagulation. PMID- 12650497 TI - Injection pain of rocuronium and vecuronium is evoked by direct activation of nociceptive nerve endings. AB - BACKGROUND AND OBJECTIVE: Rocuronium and, to a lesser extent, vecuronium can induce burning sensations associated with withdrawal reactions during administration. Dermal microdialysis in human and electrophysiological recordings of nociceptors in mouse skin were used to elucidate the underlying mechanisms of pain induction. METHODS: Microdialysis catheters were inserted intradermally into the forearm of 10 volunteers and were perfused with two different concentrations of rocuronium and vecuronium (1 and 10 mg mL(-1)) or a control. Dialysis samples were taken every 15 min and analysed for protein, histamine, tryptase and bradykinin content. Pain intensity was rated on a numerical scale of 0-10. In a parallel design, activation of cutaneous nociceptors was assessed directly in a skin-nerve in vitro preparation of the mouse hind paw. The receptive fields of identified single C-nociceptors (n = 12) were superfused with rocuronium or vecuronium solutions (10 mg mL(-1)) at physiological pH. RESULTS: In accordance with clinical observations, microdialysis of rocuronium (10 mg mL(-1)) induced sharp burning pain (NRS 4.1 +/- 1.8), whereas vecuronium given in the usual clinical concentration (1 mg mL(-1)) induced only minor pain sensations (NRS 0.6 +/- 1.3). At equimolar concentrations, pain sensation and concomitant mediator release evoked by both drugs were similar. No correlations were found between pain rating and mediator release. In the in vitro preparation, C-fibres showed a consistent excitatory response with rapid onset after stimulation with vecuronium as well as rocuronium (differences not significant). CONCLUSIONS: The algogenic effect of aminosteroidal neuromuscular blocking drugs can be attributed to a direct activation of C-nociceptors. PMID- 12650499 TI - Intrathoracic blood volume in a patient with pulmonary embolism. PMID- 12650500 TI - Prophylactic continuous intravenous ephedrine infusion for elective Caesarean section under spinal anaesthesia. PMID- 12650501 TI - Ultrasound-guided catheterization of the portal vein in 11 cows using the Seldinger technique. AB - Catheterization of the portal vein using the Seldinger technique [Acta Radiol. (1958) 38, 368] was performed in 11 cows. Ultrasound-guided percutaneous portocentesis, using a 25-cm, 14-gauge steel cannula, was performed from the 10th, 11th or 12th intercostal space on the right side. A stylet was placed through the cannula, which was then removed, and a polyurethane balloon-tipped catheter was advanced over the stylet into the portal vein and sutured to the skin (Seldinger, 1952). Blood samples were collected from the catheter at least once daily. The catheter was removed 9-15 days later when blood could no longer be aspirated. The cows were then slaughtered and a post-mortem examination was performed. During the study, appetite remained normal in nine of 11 cows. In three cows, the general behaviour and demeanour were mildly, but transiently, abnormal. Four cows had leucopoenia. The most frequently encountered problem was occlusion of the catheter, which usually was resolved by flushing with heparinized saline. The most common post-mortem lesion observed was an increase in fibrous connective tissue at the site of cannulation. In nine of 11cows, there was a thrombus in the portal vein at the site of catheterization. Generally, the severity of the lesions was mild. The results of this study demonstrated that ultrasound-guided percutaneous catheterization of the portal vein, using the Seldinger technique, is possible in cows. The catheter may be left in place for collection of blood samples for up to 15 days. PMID- 12650502 TI - The influence of a fermentation-resistant glucose diet on the glucose concentration and other metabolites in portal and jugular blood in cows. AB - The goal of this study was to determine the effect of fermentation-resistant glucose on the glucose concentration and other metabolites in portal and jugular blood in 15 non-lactating cows. In all cows, an indwelling catheter was placed in the left jugular vein and the portal vein for collection of blood samples. Five control cows were fed hay as a normal diet, five control cows were fed straw to induce an energy deficit and five cows were fed hay and they received additionally 2000 g of a fermentation-resistant D-glucose product. The glucose concentration in jugular and portal blood was not influenced by feeding. The concentration of urea and bile acids were significantly higher in portal blood than jugular blood. There was no difference between portal and jugular blood of glucose and total solids. Diet had a significant effect on the concentrations of ammonia, urea, free fatty acids and triglycerides. The concentrations of ammonia and urea were higher in blood of cows fed straw than in blood of cows fed either hay or a fermentation-resistant glucose product. The concentration of urea remained constant in cows fed hay, but increased in cows fed straw and decreased in cows fed a fermentation-resistant glucose product. The concentration of free fatty acids and triglycerides were significantly higher in cows fed a fermentation-resistant glucose product than in cows fed hay. In the present study, a single administration of 300 g of fermentation-resistant glucose did not affect the concentration of blood glucose. Therefore, despite ongoing promotion of such products, there is no indication at this time that administration of fermentation-resistant glucose to cows at the start of lactation results in an increase in blood glucose concentration. PMID- 12650503 TI - Effect of different dietary levels of Yucca schidigera powder on the performance, blood parameters and egg yolk cholesterol of laying quails. AB - A total of 135 laying quails (Coturnix coturnix japonica), 9 weeks old, were divided into three dietary treatment groups. Three replicates were assigned to each treatment group consisting of 15 birds per cage. The diet was supplemented with 0, 100 and 200 ppm Yucca schidigera powder and given ad libitum to the quails for a period of 14 weeks. Body weight, egg production, feed consumption and feed efficiency were not different due to dietary treatments among the groups. Increased egg weight was determined in the control group. Yucca powder supplementation decreased serum glucose, cholesterol triglyceride level in laying quails. Serum total protein concentration was not changed by dietary treatments but albumin level was decreased in quails fed 100 ppm yucca powder. Egg yolk cholesterol concentration was not significantly different among the groups but tended to decline (11.5%) as a result of yucca supplementation. Red Blood Cell (RBC) and White Blood Cell (WBC) counts, packed cell volume (PCV), mean corpuscular volume (MCV) and mean corpuscular haemoglobin (MCH) were not affected by supplementation of yucca powder. However, haemoglobin (HB) concentration was slightly increased and mean corpuscular haemoglobin concentration (MCHC) was significantly increased by 200 ppm yucca powder supplementation to the diet. PMID- 12650504 TI - Rumen-protected choline administration to transition cows: effects on milk production and vitamin E status. AB - We investigated the influence of rumen-protected choline (RPC) supplementation on milk production, lipid metabolism and vitamin E status in dairy cows receiving a silage-based diet. Twenty-six Italian Holstein multiparous cows were assigned by weight and average production in the previous lactation, to one of two groups: control (no RPC supplementation) and RPC (supplemented with 20 g/day rumen protected choline chloride). Treatment began 14 days before expected calving and continued for 30 days after parturition. Choline administration significantly increased milk production during the first month of lactation and also the concentration (and total secretion) of choline in milk, but did not affect fat or protein concentrations in milk, or plasma levels of glucose, beta hydroxybutyrate, cholesterol and non-esterified fatty acids (NEFA). However, around parturition, NEFA concentrations in plasma were lower in treated animals than in controls, suggesting improved lipid metabolism as a result of choline supplementation. Choline supplementation also increased alpha-tocopherol plasma concentrations, suggesting a novel aspect in dairy cows. PMID- 12650505 TI - Periparturient endocrine and metabolic changes in healthy cows and in cows affected by mastitis. AB - Transition from pregnancy to lactation in dairy cows involves considerable metabolic adaptation. Additional stress is incurred during infections such as periparturient mastitis. Multiparous Holstein-Friesian cows kept under normal production conditions (n = 15) were used to evaluate changes in circulating metabolite and hormone concentrations from 5 days before to 5 days after calving. Insulin-like growth factor binding protein (IGFBP) profiles were also monitored. Marked time-related changes were observed for plasma thyroid hormone, IGF, cortisol, insulin, beta-hydroxybutyrate (BHB) and non-esterified fatty acid (NEFA) concentrations but not for plasma leptin. A decrease in IGF-II concentration and maximal intensity of the putative IGFBP-1 band occurred at parturition. When compared with the five healthy cows,low IGF-II levels were prolonged to day 2 post-partum in five cows with Escherichia coli-associated mastitis. However, marked decreases in IGFBP-2 band intensity were evident only in two of the four cases examined. Individual total ligand (IGF-I + IGF-II) concentration and IGFBP pattern prepartum were largely regained 5 days post partum in all cows. Hormone and metabolite concentrations in the two cows with Staphylococcus aureus-associated mastitis were very similar to those in the five healthy cows. Plasma thyroxine (T4) was lower 2 days prepartum in the cows, which later developed Gram-negative mastitis. Multiregression analysis showed that variance in T4 concentration was significantly and independently associated with triiodothyronine (T3) and IGF-I positively and with cortisol negatively (R2 = 0.648). This study confirms the close inter-relationship between the thyroid hormone and IGF axes in cattle and indicates possible effects of Gram-negative mastitis infection on IGF-II metabolism. PMID- 12650506 TI - Changes in coagulation and markers of fibrinolysis in horses undergoing colic surgery. AB - Activation of coagulation can be frequently found in horses with colic. However, it has also been demonstrated as a sequela of surgical trauma alone in humans. The purpose of the present study was to determine changes in coagulation and fibrinolysis in horses that underwent colic surgery and to evaluate whether these changes were secondary to the colic or the surgery and wound healing. Thirty horses that underwent colic surgery with uncomplicated recovery were included. Ten horses with a Forssell's procedure served as control group with a standardized surgical trauma. Besides daily physical examinations during the observation period of 10 days, activated partial thromboplastin time (aPTT), prothrombin time and thrombin time as well as fibrin monomer (FM), D-Dimer (DD) and antithrombin (AT) III were determined. Compared with the control group the aPTT was the only standard coagulation test that was significantly prolonged before and after the event of colic surgery. After surgery, hyperfibrinogenaemia occurred in all groups. In colic groups FM and DD concentrations were within reference range at admission,and were significantly greater than in control horses after surgery. AT III activity decreased after colic surgery, but did not change in the control group. It was concluded that an activated coagulation state after colic surgery has to be expected, resulting not only from the colic disease, but also from the event of surgery. PMID- 12650507 TI - Day-to-day variability in glomerular filtration rate in normal dogs by scintigraphic technique. AB - The sources of variability in variability of scintigraphic measurements of glomerular filtration rate (GFR) have not been determined. The day to day variability of GFR was studied in 18 healthy beagle dogs. The renal uptake of 99mTc-diethylenetriaminepentaacetic acid (DTPA) of each dog was measured using a scintigraphic technique three times at intervals of 5-26 days. GFR was calculated from a regression equation relating uptake to plasma clearance, derived in our laboratory. The mean GFR was 3.97 +/- 0.72 (SD) ml/min/kg with values from 2.66 to 5.67 ml/min/kg. Analysis of variance (ANOVA) using a linear mixed model showed that most variability is a result of the dogs, less because of day to day variability and very little to the measurement variability. The repeatability coefficients for the day to day variability and measurement variability were 1.06 and 0.21 ml/min/kg respectively. The day to day variability can be caused by physiological homeostatic adjustments by the kidneys needed because of fluctuations in food and fluid intake, each dog's individual capacity to adjust, and to intrinsic errors in the measurement method. These results should be considered when using the scintigraphic method for clinical evaluation and research. PMID- 12650509 TI - BSE surveillance in Italy: neuropathological findings in cattle in the frame of the passive surveillance programme. PMID- 12650508 TI - Equine third metacarpal bone assessment by quantitative ultrasound and dual energy X-ray absorptiometry: an ex vivo study. AB - The purpose of this ex vivo study was to analyse two commonly established methods of mechanical bone property assessment for application in horses: Quantitative ultrasound (QUS), which depends on the bone's density and Young's modulus, and dual energy X-ray absorptiometry (DXA), which depends on the areal bone mineral density (BMD). The third metacarpal bone (MC III) of horses was selected as examination region for practical reasons. An interrelationship between QUS- and DXA-values was examined. Both MC III of eleven randomly selected equine cadavers were divided in nine regions of interest (ROI). A multi-site QUS device was used for axial transmission speed of sound (SOS) measurements and a DXA device was used for BMD evaluation. Full cortical thickness BMD (FcBMD), overall aspect BMD and 4 mm cortical border slice BMD (4 mmBMD) were evaluated. In addition, each ROI of one MC III was measured 10 times to determine QUS- and DXA-measurement precision. SOS values and BMD values obtained at different aspects of MC III were different (P < 0.001). FcBMD and overall BMD obtained at different levels were different (P < 0.001). SOS data correlated with FcBMD-, overall BMD- and 4 mmBMD data at various ROI. FcBMD-, overall BMD- and 4 mmBMD-data were strongly correlated. The intra-operator coefficient of variation was 1.3% for SOS measurements and ranged between 1.94 and 10.3% for BMD-measurements. Multi-site axial transmission QUS as well as DXA can be used to precisely measure bone characteristics of MC III in horses. However, both techniques do not measure the same bone properties. It is therefore concluded, that QUS and DXA techniques are complementary for application in horses. PMID- 12650510 TI - Fatal systemic phaeohyphomycosis in a cat due to Cladophialophora bantiana. AB - We report a case of fatal systemic mycosis in a 9-year-old cat. Diagnosis of phaeohyphomycosis was made by histology. Morphological and molecular identification of the fungus isolated from the lesions yielded the species Cladophialophora bantiana. This species is well known to be a neurotropic fungus causing cerebral pyogranulomatous lesions in humans. In this case, the lesions were widespread, distributed without the involvement of central nervous system. The origin of systemic manifestation is still unknown and no evidence of immunosuppression was found. It is the first feline case of C. bantiana infection reported in Europe. PMID- 12650511 TI - Intramucosal Helicobacter pylori in the human and murine stomach: its relationship to the inflammatory reaction in human Helicobacter pylori gastritis. AB - Intramucosal Helicobacter pylori (H. pylori) has been described in biopsy tissues and culture systems. However, the association of intramucosal H. pylori with histopathologic features has not been evaluated. The purpose of this study is to investigate the relationship between intramucosal H. pylori and inflammatory reactions in H. pylori infection. In 113 randomly selected human gastric biopsies and 20 murine stomachs, which were inoculated with SSI every day for a week, immunohistochemical analysis for intramucosal H. pylori was done and correlated with histologic parameters. Electron microscopic examination was done on murine stomachs. H. pylori infection was present in 104 gastric biopsies and 17 murine stomachs. Intraepithelial immunopositivity for H. pylori was detected in 27 of 104 (26%) biopsies and in 11 of 17 (65%) murine stomachs. Lamina proprial immunopositivity for H. pylori was present in 51 of 104 (48%) biopsies. Neutrophil-associated immunopositivity for H. pylori was observed in 22 of 90 (24%) biopsies with H. pylori chronic active gastritis. Lamina proprial and neutrophil-associated immunopositivity for H. pylori correlated significantly with the density of H. pylori and the grade of acute inflammatory reaction in H. pylori gastritis. Intramucosal location of H. pylori itself or its antigen is closely associated with acute inflammatory reactions and may play an important role in establishing a persistent infection in chronic H. pylori gastritis. Furthermore, lamina proprial and/or neutrophil-associated H. pylori appears to be more important than intraepithelial H. pylori in acute inflammatory reactions of H. pylori gastritis. PMID- 12650512 TI - Clinical and autopsy correlation evaluated in a university hospital in Spain (1991-2000). AB - The authors present a 10-year retrospective study (1991-2000) of all autopsies performed at the Hospital Clinic, Barcelona (Catalonia, Spain) studied by a multidisciplinary committee. The clinicopathologic correlation of the primary underlying disease with the immediate cause of death was reviewed. Between 1991 and 2000, 2,495 autopsies were performed, 1933 of which were evaluated by the committee. The autopsy rate fell from 20% in 1993 to 9.1% in 2000. The clinicopathologic correlation in underlying primary disease was correct in 92.67% of the cases; there was a major discrepancy in 3.51% and a minor discrepancy in 3.82%. As regards the immediate cause of death, major errors were found in 5.89% of cases and minor errors in 6.17%. Despite the scientific and technologic advances in medicine, we have seen that there are still clinicopathologic discrepancies. The postmortem examination continues to play an important role in auditing clinical practice and diagnostic performance, and also for educational purposes. Evaluation by a multidisciplinary committee is the more reliable system for the study of the clinicopathologic correlation. PMID- 12650513 TI - Case control study of novel prognostic markers and disease outcome in pregnancy/lactation-associated breast carcinoma. AB - A case control study of pregnancy/lactation associated breast carcinoma (PAC) was conducted on 24 test cases with two controls per case, matching age, tumor grade, tumor size and axillary lymph nodes status. During seven years of this study, 6% of all patients with breast cancer had PAC. In this study, 67% of the test cases showed positive axillary lymph nodes compared to 49% in our series of 315 cases of non-pregnancy/non-lactating women with breast carcinoma (p < 0.05). The expression of nine prognostic markers, i.e. ER, PR, p53, C-erbB-2, EGFR, Cathepsin-D, PCNA, DNA ploidy and S-phase fraction, were studied by immunohistochemistry and flow cytometry. Hormone receptor status showed a statistically significant difference between the two groups, i.e. 29% immunoreactivity in test cases compared to 58% in controls with a p value of 0.007. Among p53, C-erbB-2, EGFR and Cathepsin-D in the test group, only EGFR showed a significant correlation, i.e. 33% immunoreactivity in test cases and 19% immunoreactivity in controls (p < 0.05). Higher PCNA positivity was seen in the test group compared to controls, i.e. 35% in test patients and 28% in controls (p < 0.05). Metastasis to bone and liver was a common feature of test patients as compared to controls (p < 0.05). After a median follow-up of 72 months, there was no significant difference in the overall survival (OS) of test cases and controls as 54% deaths were recorded in test patients and 44% in controls at the end of this study (p > 0.05). In summary, in spite of some significant differences in the expression of few prognostic markers, i.e. ER/PR, EGFR, PCNA and metastatic potential, there was no significant difference in the OS of PAC vs. control group if compared stage for stage. PMID- 12650514 TI - Expression of transcription factor E2F-1 in pancreatic ductal carcinoma: an immunohistochemical study. AB - E2F-1 is a transcriptional factor that mediates cell cycle progression from G1 to S phase, thereby influencing tumor progression. However, only a few clinicopathologic studies have been carried out using surgically removed specimens for defining its role in tumor biology. Therefore, we studied the expression of this cell cycle regulator on surgical specimens at the immunohistochemical level, and examined its possible relationship with proliferative index, assessed by analysis of MIB-1 expression, and clinicopathologic factors in pancreatic ductal carcinomas. E2F-1 and MIB-1 were immunostained on 54 surgically removed specimens, and nuclear reactivity was evaluated. The percentage of E2F-1 positive cells (E2F-1 PI) ranged from 3.8% to 71.4%. We found a statistically significant correlation between E2F-1 PI and the histologic grade of tumor differentiation (p = 0.0133), i.e. E2F-1 PI was higher in less-differentiated carcinomas. Furthermore, there was a positive correlation between E2F-1 PI and the percentage of MIB-1 PI (r = 0.763; p < 0.0001). The patients with higher E2F-1 PI (E2F-1 PI > or = 38.0 = median) showed a significantly shorter disease-associated survival time in R0 resection cases (n = 49, p = 0.015). The present analysis seems to support the theory that E2F-1 is upregulated in cell cycle, and its expression reflects the effector function of G1/S progression as far as pancreatic ductal carcinoma is concerned. PMID- 12650515 TI - PAS inclusions, immunoreactive tenascin and proliferative activity in low-grade chondrosarcomas. AB - To distinguish between chondrosarcoma (grade 1--borderline histology) and enchondroma, we examined six chondrosarcomas (grade 1--borderline histology) which looked like benign lesions. Their diagnosis, albeit based on clinical, radiologic and pathologic examinations, was not easily reached. Moreover, we examined six enchondromas and 11 chondrosarcomas, the diagnoses of which were straightforward. All cartilaginous tumors were studied, placing emphasis on PAS positive intracytoplasmic globules. Anti-Ki67 proliferation-associated nuclear antigen antibody and tenascin antibody were applied. The following features were observed in low-grade chondrosarcomas: (1) masses of hyalin and/or myxoid cartilage invading spaces around the tumor, (2) host lamellar bone trabeculae surrounded by cartilage on all sides, (3) tumoral resorption of bone trabeculae. Intracytopasmic hyalin globules (ICG) were more frequently found in malignant than in benign neoplasm (p = 0.042). Moreover, tenascin matrix immunoreactivity was more likely to be observed in benign than in malignant neoplasm (p = 0.029). Ki67 immunoreactivity was more frequent in characterized than in low-grade chondrosarcomas or in enchondromas, where it was null (p = 0.0044). PMID- 12650516 TI - Cellular mesoblastic nephroma: morphologic, cytogenetic and molecular links with congenital fibrosarcoma. AB - Congenital mesoblastic nephroma (CMN) is a rare renal tumor of early infancy with a favorable outcome after complete surgical removal. CMN consists of a heterogeneous group of spindle cell tumors subdivided into "classical", "cellular or atypical" and "mixed" forms based on histologic features. We describe a new case of cellular CMN diagnosed by antenatal ultrasonography with complete remission five years after nephrectomy. Cytogenetic study evidenced a trisomy 11, and real time RT-PCR, but not conventional karyotype, allowed for the detection of the Tel-ETV6/TrkC-NTRK3 fusion transcript as a consequence of a cryptic t(12 15)(p13;q25). As in congenital fibrosarcoma (CFS), two Tel-ETV6/ TrkC-NTRK3 fusion transcripts different by a 42 bp insert in the TrkC kinase domain were expressed. Our observations outline the close links between cellular CMN and CFS. Both tumors have the clinical presentation and histologic features as well as identical cytogenetic and molecular markers in common. Therefore, they are likely to represent the same neoplasm, but occurring at different locations. PMID- 12650517 TI - Inflammatory leiomyosarcoma of the retroperitoneum. AB - We present a case of an inflammatory leiomyosarcoma, a variant of smooth muscle cell tumor that could be mistaken for inflammatory malignant fibrous histiocytoma. The inflammatory cell infiltrates obscured the basic architecture of the tumor, and immunohistochemistry was required to arrive at the proper diagnosis. Since inflammatory leiomyosarcomas are rare and possibly misdiagnosed neoplasms, there are not enough data to indicate whether they deserve to be separated from other malignant smooth muscle cell neoplasms. By reporting this case we hope that more pathologists will become aware of this tumor type and that more cases will be collected for future clinicopathologic studies. PMID- 12650518 TI - Perivascular epithelioid cell tumor of the jejunum. AB - Certain HMB-45-positive epithelioid cell tumors have recently been categorized under a unified concept: perivascular epithelioid cell tumor (PEComa). In this report, we describe ajejunal PEComa arising in a 32-year-old woman without other tumors or stigmata of tuberous sclerosis. The tumor consisted of nests of epithelioid cells with clear to granular eosinophilic cytoplasm. The nests were separated by thin fibrovascular septa. The tumor cells were positive for HMB-45 and progesterone receptor, and negative for cytokeratin, epithelial membrane antigen, vimentin, desmin, alpha-smooth muscle actin and CD34. RT-PCR analysis failed to reveal fusion transcript ETW/ATF1, which is characteristic of clear cell sarcoma of the soft parts. She developed a recurrent tumor at the pelvic wall and the left ovary at 13 and 25 months after the first operation, respectively. Each tumor was resected surgically, and no additional therapy was performed. We think the tumor of this case is a malignant form of PEComa because of the clinical history of multiple recurrences and the size of the primary tumor. Our case underscores that to make a correct diagnosis, clinical information and immunohistochemical examination are essential. PMID- 12650519 TI - Hemangioma of the umbilical cord: report of a case. AB - A 31-year-old woman with a large placental tumor underwent a caesarean section. After delivery, the lesion, detected by ultrasound examination, was found to originate from the umbilical cord. A live female infant with cyanosis, hypotonia and diffuse edema was delivered. A review of the literature, which revealed 31 cases of umbilical cord hemangioma, showed that this tumor has a polymorphous presentation. Some fetuses and infants died from various causes, indicating that a close follow-up is necessary in these pregnancies PMID- 12650520 TI - Elevation of transgene expression level by flanking matrix attachment regions (MAR) is promoter dependent: a study of the interactions of six promoters with the RB7 3' MAR. AB - We have analyzed effects of a matrix attachment region (MAR) from the tobacco RB7 gene on transgene expression from six different promoters in stably transformed tobacco cell cultures. The presence of MARs flanking the transgene increased expression of constructs based on the constitutive CaMV 35S, NOS, and OCS promoters. Expression from an induced heat shock promoter was also increased and MARs did not cause expression in the absence of heat shock. There was also no effect of MARs on the pea ferredoxin promoter, which is not normally expressed in this cell line. Importantly, most transgenes flanked by RB7 MAR elements showed a large reduction in the number of low expressing GUS transformants relative to control constructs without MARs. PMID- 12650521 TI - Enforced expression of oxygen-regulated protein, ORP150, induces vacuolar degeneration in mouse myocardium. AB - Although the 150 kDa oxygen-regulated protein (ORP150) is known as a protein induced by low oxygen tension or ischemical insult, its possible role has not been fully investigated in vivo. To investigate the intracellular function of this protein, we generated the ORP150 over-expressing transgenic mice (ORP-Tg mice) under beta-actin promoter, and established three independent lines of the transgene expressed mice. All lines invariably showed growth retardation. Over expression of ORP150 was confirmed by western blotting in heart, brain, spleen, skeletal muscle, pancreas, lung, thymus, and kidney. To ascertain the relationship between the over-expression of the ORP150 and the growth retardation in the transgenic mice, we examined pathological changes in the transgenics. In the ORP-Tg mice, vacuolar degeneration appeared in the heart. The degeneration in the myocytes became conspicuous with advancing age. Immunostaining demonstrated ORP150 in the vacuoles of degenerating myocytes. Electron microscopical findings revealed striking development of intracellular membrane system, for example, rough endoplasmic reticula (rER), vacuoles and Golgi bodies, swelling of sarcoplasmic reticulum, and lysis of myofibrils and mitochondria. These findings indicate that ORP150 may locate in the rER and other outer compartment of ER, and that constitutive over-expression of ORP150 in the heart induces vacuolar degeneration in myocytes, resulting in growth retardation of the transgenics. PMID- 12650522 TI - Transgenic expression of trypsin inhibitor CMe from barley in indica and japonica rice, confers resistance to the rice weevil Sitophilus oryzae. AB - Indica and japonica rice (Oryza sativa L.) plants were transformed by particle bombardment with the Itr1 gene encoding the barley trypsin inhibitor BTI-CMe, under the control of its own promoter that confers endosperm specificity, and the maize ubiquitin promoter. From 38 independent transgenic lines of indica (breeding line IR58) and 15 of the japonica (cv Senia) selected, 22 and 11, respectively, expressed the barley inhibitor at detectable levels. The transgene was correctly translated as indicated by western blot analysis with a level of expression in R3 seeds up to 0.31% (IR58) and 0.43% (Senia) of the total extracted protein. The functional integrity of BTI-CMe was confirmed by trypsin activity assays in liquid media and by activity staining gels, performed with seed extracts. The significant reduction of the survival rate of the rice weevil (Sitophilus oryzae, Coleoptera: Curculionidae) reared on homozygous transgenic indica and japonica rice seeds expressing the BTI-CMe, compared to non transformed controls, and the decrease in the trypsin-like activity of insect crude midgut extracts, confirmed the utility of this proteinase inhibitor gene for the control of important storage pests. PMID- 12650523 TI - Tetracycline-inducible system for regulation of skeletal muscle-specific gene expression in transgenic mice. AB - Tightly regulated control of over-expression is often necessary to study one aspect or time point of gene function and, in transgenesis, may help to avoid lethal effects and complications caused by ubiquitous over-expression. We have utilized the benefits of an optimized tet-on system and a modified muscle creatine kinase (MCK) promoter to generate a skeletal muscle-specific, doxycycline (Dox) controlled over-expression system in transgenic mice. A DNA construct was generated in which the codon optimized reverse tetracycline transactivator (rtTA) was placed under control of a skeletal muscle-specific version of the mouse MCK promoter. Transgenic mice containing this construct expressed rtTA almost exclusively in skeletal muscles. These mice were crossed to a second transgenic line containing a bi-directional promoter centered on a tet responder element driving both a luciferase reporter gene and a tagged gene of interest; in this case the calpain inhibitor calpastatin. Compound hemizygous mice showed high level, Dox dependent muscle-specific luciferase activity often exceeding 10,000-fold over non-muscle tissues of the same mouse. Western and immunocytochemical analysis demonstrated similar Dox dependent muscle-specific induction of the tagged calpastatin protein. These findings demonstrate the effectiveness and flexibility of the tet-on system to provide a tightly regulated over-expression system in adult skeletal muscle. The MCKrtTA transgenic lines can be combined with other transgenic responder lines for skeletal muscle-specific over-expression of any target gene of interest. PMID- 12650524 TI - A self-excising Cre recombinase allows efficient recombination of multiple ectopic heterospecific lox sites in transgenic tobacco. AB - To study the impact of different DNA configurations on the stability of transgene expression, a variant of the cre gene was developed. This variant allows for the highly efficient in planta removal of its own loxP-flanked coding sequence as well as other DNAs flanked by ectopic heterospecific lox sites, either lox511 or lox2272 or both, in trans. The plant intron-containing cre gene, creINT, was configured in such a way that self-excision generated an intact hygromycin resistance selectable marker gene. In this combination, all selected transformants showed highly efficient excision. Plants obtained showed no indication of any chimerism, indicating a cell autonomous nature of the hygromycin selection during transformation and regeneration. The highly efficient concomitant removal of wildtype and heterospecific lox site-flanked DNA demonstrated that upon retransformation with the self-excising creINT, sufficient amounts of Cre enzyme were produced prior to its removal. Plants obtained with creINT showed much less frequently the Cre-associated phenomenon of reduced fertility than plants obtained with a continuous presence of Cre recombinase. The creINT system has therefore advantages over systems with a continuously present Cre. The creINT system was successfully used for removal of two chromatin boundary elements from transgene cassettes in tobacco. Analysis of plants with and without boundary elements on the same chromosomal location will contribute to a better evaluation of the role of such elements in the regulation of transgene expression in plants. PMID- 12650525 TI - Strain-dependent differences in the efficiency of transgenic mouse production. AB - Transgenic mouse production via pronuclear microinjection is a complex process consisting of a number of sequential steps. Many different factors contribute to the effectiveness of each step and thus influence the overall efficiency of transgenic mouse production. The response of egg donor females to superovulation, the fertilization rate, egg survival after injection, ability of manipulated embryos to implant and develop to term, and concentration and purity of the injected DNA all contribute to transgenic production efficiency. We evaluated and compared the efficiency of transgenic mouse production using four different egg donor mouse strains: B6D2/F1 hybrids, Swiss Webster (SW) outbred, and inbred FVB/N and C57BL/6. The data included experiments involving approximately 350 DNA transgene constructs performed by a high capacity core transgenic mouse facility. Significant influences of particular genetic backgrounds on the efficiency of different steps of the production process were found. Except for egg production, FVB/N mice consistently produced the highest efficiency of transgenic mouse production at each step of the process. B6D2/F2 hybrid eggs are also quite efficient, but lyze more frequently than FVB/N eggs after DNA microinjection. SW eggs on the other hand block at the 1-cell stage more often than eggs from the other strains. Finally, using C57BL/6 eggs the main limiting factor is that the fetuses derived from injected eggs do not develop to term as often as the other strains. Based on our studies, the procedure for transgenic mouse production can be modified for each egg donor strain in order to overcome any deficiencies, and thus to increase the overall efficiency of transgenic mouse production. PMID- 12650526 TI - Stable inheritance of the antisense Waxy gene in transgenic rice with reduced amylose level and improved quality. AB - Amylose content in rice endosperm is a key determinant of eating and cooking quality. In the present study, a chimeric antisense construct, which contained a 756-bp antisense Waxy (Wx) gene DNA fragment from rice and the gusA coding sequence, both fused to the 3.1-kb rice Wx promoter, was efficiently introduced into several elite rice cultivars, both of japonica and indica type, via Agrobacterium. More than 200 independent transgenic lines were produced and integration transgene was confirmed by PCR and Southern blotting. Northern blot analysis suggested that the antisense Wx transcript interacted with both the endogenous Wx mature mRNA and unspliced transcripts, but only interaction with the mature mRNA resulted in reduced amylose synthesis. Analysis of GUS activity showed that the gusA fusion gene driven by the rice Wx promoter expressed highly in the endosperm of the transgenic rice plants. Varying degrees of reduction in amylose content, up to 96%, were found in seeds derived from these transformants. Consistently, opaque white seeds, similar to glutinous rice, were observed in several transgenic lines of japonica rice. In transgenic lines derived from indica rice, which usually has a high amylose level, significant reduction of amylose content was also found in the endosperm, but the levels of reduction were lower than those of japonica rice. Genetic analysis demonstrated that transgenes and improved amylose content were stably inherited (up to ninth generation) in these transgenic lines. Several elite transgenic lines with improved amylose level and quality have been selected for field evaluation. PMID- 12650527 TI - Molecular structure and regulatory potential of a T-DNA integration site in petunia. AB - The genomic structure surrounding a T-DNA integration site in a transgenic petunia plant, which shows deregulation of a root-specific promoter, was investigated. We have already demonstrated that T-DNA integration in this transformant (P13) had occurred close to a scaffold/matrix attachment region (S/MAR). A major question regarding the observed promoter leakiness was whether the T-DNA had integrated into the centre or at the border of the Petun-SAR and whether other regulatory elements are located within this genomic region. While small rearrangements were shown to occur during T-DNA integration in agreement with other reports, we find indications of the presence of a SINE retroposon--an apparent landmark for recombinogenic targets--at the integration site. Binding assays to both plant and animal nuclear scaffolds, supported by biomathematical analyses, reveal that the T-DNA is definitely located at the border of a strong S/MAR, which is in agreement with current models on the structure of integration sites. These results, together with a developmentally regulated leaf-specific enhancer effect of the Petun-SAR on gene expression in transgenic tobacco plants, indicate that the Petun-SAR demarcates the right border of a chromatin domain with genes predominantly active in leaves. PMID- 12650529 TI - Chloroplast transformation in oilseed rape. AB - The chloroplast transformation vector pNRAB carries two expression cassettes for the spectinomycin resistance gene aadA and the insect resistance gene cry1Aa10. The two cassettes are sited between the rps7 and ndhB targeting fragments. Biolistic delivery of the vector DNA, followed by spectinomycin selection, yielded chloroplast transformants at a frequency of four in 1000 bombarded cotyledon petioles. PCR analysis and Southern blot of PCR products confirmed the site-specific integration of aadA and cry1Aa10 into the chloroplast genomes of transgenic oilseed rape. When transgenic oilseed rape leaves were fed to second instar Plutella xylostera larvae, 47% mortality was observed against this insect and the surviving larvae had significantly lower weight than the control. This is the first report of chloroplast transformation in oilseed rape and the introduction of novel genes between the rps7 and ndhB genes in the chloroplast genome. This offers an opportunity for improvement of oilseed rape by chloroplast genetic engineering. PMID- 12650528 TI - Chemical-inducible, ecdysone receptor-based gene expression system for plants. AB - We have developed an inducible gene expression system with potential for field application using the ecdysone receptor (EcR) from the spruce budworm and the non steroidal EcR agonist, methoxyfenozide. Chimeric transcription activators were constructed with EcR ligand binding domain, GAL4 and LexA DNA binding domains, and VP16 activation domain. In the presence of methoxyfenozide, the transcription activators induced expression of the luciferase reporter gene cloned downstream of a promoter containing GAL4A- or LexA-response element and a minimal 35S promoter. Low basal and high induced luciferase expression was optimized by cloning the activator and the reporter genes in different tandem orientations. Many transgenic Arabidopsis and tobacco plants were obtained with little or no basal expression in the absence of methoxyfenozide and inducible expression that was several fold higher than that observed with the constitutive 35S promoter. Moreover, gene expression was controlled over a wide range of methoxyfenozide concentration. Our results demonstrate that the inducible gene expression system based on the spruce budworm EcR ligand binding domain with methoxyfenozide as a ligand is very effective in regulating transgenes in plants. It is suitable for field applications because methoxyfenozide is commercially available and has an exceptional health and environmental safety profile. PMID- 12650530 TI - Plastid transformation in Lesquerella fendleri, an oilseed Brassicacea. AB - A plastid transformation protocol was developed for Lesquerella fendleri, a species with a high capacity for plant regeneration in tissue culture. Transformation vector pZS391B carried an aadA16gfp marker gene conferring streptomycin-spectinomycin resistance and green fluorescence under UV light. Biolistic transformation of 51 Lesquerella leaf samples, followed by spectinomycin selection, yielded two transplastomic clones. The AAD-GFP fusion protein, the marker gene product, was localized to chloroplasts by confocal laser microscopy. Fertile plants and seed progeny were obtained in line Lf-pZS391B-1. In the 51 samples a large number (108) of spontaneous mutants were identified. In five of the lines spectinomycin resistance was localized to a conserved stem structure by sequencing 16S rRNA genes. Success in L. fendleri, a wild oilseed species, extends plastid transformation beyond Arabidopsis thaliana in the Brassicaceae family. PMID- 12650531 TI - Coexpression of the soybean vegetative storage protein beta subunit (S-VSPbeta) either with the bacterial feedback-insensitive dihydrodipicolinate synthase or with S-VSPalpha stabilizes the S-VSPbeta transgene protein and enhances lysine production in transgenic tobacco plants. AB - Soybean vegetative storage proteins (S-VSPs) are lysine-rich leaf proteins, originally found to accumulate to high levels in depodded soybean plants. In the present study, we overexpressed S-VSPbeta, the ruminant stable subunit of the S VSP genes, in transgenic tobacco plants. The S-VSPbeta protein accumulated in all organs studied, but its level declined drastically with leaf age. This instability of S-VSPbeta could be overcome either by elevating free lysine levels or by coexpressing S-VSPbeta with S-VSPbeta. High levels of rumen-stable, lysine rich proteins is expected to improve absorption of lysine by ruminants. Furthermore, the expression of S-VSPs in heterologous plants led to a significant increase in total soluble lysine, suggesting that these proteins may also permit better assimilation of lysine by humans and monogastric animals. PMID- 12650532 TI - Oral immunization using tuber extracts from transgenic potato plants expressing rabbit hemorrhagic disease virus capsid protein. AB - Rabbit hemorrhagic disease, which is caused by a calicivirus, is a lethal infection of adult animals that is characterized by acute liver damage and disseminated intravascular coagulation. In this study, we report the production of the major structural protein VP60 of rabbit hemorrhagic disease virus in transgenic tubers of potato plants and its use as an oral immunogen in rabbits. PMID- 12650533 TI - The d subunit of the vacuolar ATPase (Atp6d) is essential for embryonic development. PMID- 12650534 TI - Where did they all go? PMID- 12650535 TI - Fat embolism syndrome. AB - Fat embolization and the clinical syndrome associated with this pathology are poorly understood complications of skeletal trauma. Fat embolization is characterized by release of fat droplets into systemic circulation after a traumatic event. Fat embolism syndrome (FES) is an infrequent clinical consequence of fat embolization. Classically, FES presents with the triad of pulmonary distress, mental status changes, and petechial rash 24 to 48 hours after pelvic or long-bone fracture. FES incidence increases with the number of fractures sustained by an individual. Many clinicians believe that FES incidence has decreased over the past several decades secondary to advances in resuscitative measures. FES pathophysiology remains unclear. Current theories involve common mechanical and biochemical mechanisms that explain how fat emboli manifest as FES. Much controversy surrounds the question of whether there is a causal relation between intramedullary nailing and FES onset. Clinical diagnosis is essential, as laboratory and radiographic findings are nonspecific. Early supportive pulmonary therapy and other resuscitative measures may halt the pathophysiologic cascade and prevent clinical deterioration. Fortunately, if FES is diagnosed early, and pulmonary and cardiac functions are optimally supported, prognosis is very good. PMID- 12650536 TI - Blood transfusion in proximal humerus fractures. AB - This article is the first to specifically address blood transfusions associated with surgical treatment of proximal humerus fractures. In our study, we retrospectively reviewed charts of all patients admitted to our institution with a diagnosis of proximal humerus fracture from January 1, 1994, to December 31, 1997. The transfusion rate for patients who sustained a proximal humerus fracture was 15%. Compared with patients treated nonoperatively, patients who underwent a surgical procedure were not at increased risk for requiring transfusion. Age, hematocrit level on admission, treatment method, and estimated amount of blood loss in patients who underwent a surgical procedure varied significantly between those who received a transfusion and those who did not. Certain characteristics placed patients at increased risk for requiring transfusion--age older than 60, admission hematocrit level less than 33%, and hemiarthroplasty as definitive surgical treatment. This information is important both in educating patients and their families and in making management decisions. PMID- 12650537 TI - Deleterious effect of smoking on healing of open tibia-shaft fractures. AB - We retrospectively reviewed a prospectively followed cohort of 105 patients with 110 open tibia fractures treated with external fixator or intramedullary nail to determine whether smoking affects fracture healing. Severe open tibia-shaft fractures treated at a tertiary-care medical center were included. Patients with type II, IIIA, or IIIB tibia fractures were eligible. Treatment for all patients was similar, except that they were randomized to receive external fixator or intramedullary nail. Time to fracture healing was the main outcome measurement. Smokers had a union rate of 84% (52/62), and nonsmokers had a union rate of 94% (45/48), P = .10. For smokers in one arm of the study, time to union was significantly longer (P = .01), and there were more complications (P = .04). Smoking decreased unions, slowed healing, and increased complications. PMID- 12650538 TI - Secretan's disease: a unique case report and literature review. AB - Secretan's disease was first described in 1901 as "hard edema and traumatic hyperplasia of the dorsum of the metacarpus." Other names for this process include peritendinous fibrosis, posttraumatic hard edema, and factitious lymphedema. Over the years, many theories have been proposed as to the etiology of the disease--trauma that leads to peritendinous fibrosis, hypersympathetic stimulus after an injury, and self-inflicted trauma. There is no consensus regarding pathophysiology, classification, prognosis, or treatment, but the literature suggests that Secretan's disease is an injury that is self-inflicted either for secondary gain or as a conversion reaction and that is best treated with conservative care and psychiatric counseling. Our case report is unique in that our patient had an unusual presentation, underwent an upper extremity sympathectomy before developing Secretan's disease, and returned to work soon after surgery. PMID- 12650539 TI - Remodeling potential for fracture deformity of the proximal phalanx of the finger in children. AB - We report the case of a 28-month-old patient with a grossly deformed fracture malunion of the finger. The malunion was remodeled completely without any intervention. PMID- 12650540 TI - Tibiotalocalcaneal fusion with a retrograde intramedullary nail: clinical and functional outcomes. AB - Severe arthrosis and deformity of the ankle and subtalar joints are debilitating problems that can be difficult to treat. We retrospectively reviewed and functionally assessed 15 patients who had undergone tibiotalocalcaneal fusion with a retrograde intramedullary nail. The procedure had been a salvage procedure for severe arthrosis and deformity of the ankle and subtalar joints. Minimum follow-up was 2 years. Solid fusion was achieved in 14 of 15 patients. Mean time to union was 16.5 weeks. Ankle-Hindfoot Scale scores showed excellent pain relief at union. Subjective gait disturbances were common postoperatively, though patient satisfaction and the return-to-work rate were high (93% and 80%, respectively). The 12 complications included nonunion, malunion, skin problems, plantar pain, infection, and painful hardware. We advocate using a retrograde nail as a salvage technique in complex posttraumatic or postsurgical settings. PMID- 12650541 TI - Use of a stockinette in closed reduction of distal extra-articular tibia fractures. AB - Tibia fractures are common orthopaedic injuries. One of the most difficult tibia fractures that physicians encounter is the extra-articular distal third fracture. Operative fixation has certain risks depending on the procedure. Intramedullary nails fail to obtain adequate distal locking, and plate fixation increases the risk of soft tissue complications. Therefore, when indicated, closed reduction should be attempted. We have devised a method of applying in-line traction while performing closed reduction and casting with the use of a stockinette. This method avoids many of the problems encountered with other techniques such as calcaneal traction pins and hair splints. With the use of in-line traction through the stockinette, we are able to apply traction throughout casting without assistance, and the procedure is noninvasive. PMID- 12650543 TI - Field-portable spectroscopy. PMID- 12650542 TI - Hazards associated with the manufacture and repair of neon lights. PMID- 12650544 TI - Development of a hydraulic crimping tool. PMID- 12650545 TI - Overexposure to crystalline silica in a foundry operation. PMID- 12650546 TI - Diesel exhaust exposure in the Canadian railroad work environment. AB - An investigation of occupational exposure to diesel exhaust, in terms of elemental carbon, was conducted as part of a feasibility study in the Canadian railroad industry. Both personal and area samples were collected from three major operating divisions of the railways: mechanical service, transportation, and engineering. A total of 255 elemental carbon samples have been described. The results show that all but six elemental carbon concentrations, expressed as size selective respirable air samples taken using a 10 mm nylon cyclone, are well below the 2001 proposed American Conference of Governmental Industrial Hygienists' (ACGIH) threshold limit value (TLV) of 20 microg/m3. The concentration of diesel exhaust, expressed as elemental carbon, in the railroad industry is much lower than that in some other major industries such as mining and forklift truck operations. If the TLV is to be applicable to a broad range of workplace settings such as railroad, construction, and mining, the use of a TLV that is based on an elemental carbon measurement of size selective respirable samples, as recommended in the 2001 ACGIH proposal, would appear to be the most valid strategy for control of exposure to diesel exhaust. PMID- 12650547 TI - Short daily exposure to hand-arm vibrations in Swedish car mechanics. AB - The aim of the study was to examine the daily exposure times to hand-arm vibrations in Swedish car mechanics, to test a method for estimating the exposure time without observing the workers for whole days, and to use the results for predicting the prevalence of vibration-induced white fingers (VWF) by the ISO 5349-model. Six garages were surveyed. In each garage, 5-10 car mechanics were observed in random order every 30 seconds throughout working days. The daily exposure time for each mechanic was estimated from the fraction of the observations that the mechanic was exposed. A total of 51 mechanics were observed, most of them on two different working days, yielding estimates for 95 days. The median effective exposure time was 10 minutes per day (95% confidence interval 5-15 minutes, arithmetic mean 14 minutes, maximum 80 minutes), and most of the exposure time was attributable to fastening and loosening nuts. The within worker and between-worker variability was high (total sigma2 0.99, geometric standard deviation of 2.7). Using the observed exposure time and data on vibration levels of the main tools in Swedish car mechanics (average weighted acceleration level of 3.5 m/s2), the model in ISO-standard 5349 would predict that only three percent of the car mechanics will suffer from VWF after 20 years of exposure. In contrast, a recent survey of VWF showed the prevalence to be 25 percent. The precision of the observation method was estimated and was found to be good for the group daily mean. On the individual level the precision was only acceptable if the daily exposure time was > or = 40 minutes. In conclusion, the daily exposure time was short and the vibration level was limited. Nevertheless, hand-arm vibrations cause VWF in a significant number of car mechanics. The method of observing workers intermittently seemed to work well. PMID- 12650548 TI - Rapid detection and determination of the aerodynamic size range of airborne mycobacteria associated with whirlpools. AB - Novel environmental air and water mycobacteria sampling and analytical methods are needed to circumvent difficulties associated with the use of culture-based methodologies. To implement this objective, a commercial, clinical, genus DNA amplification method utilizing the polymerase chain reaction (PCR) was interfaced with novel air sampling strategies in the laboratory. Two types of air samplers, a three-piece plastic, disposable filter cassette and an eight-stage micro orifice uniform deposit impactor (MOUDI), were used in these studies. In both samplers, 37-mm polytetrafluoroethylene (PTFE) filters were used. Use of the MOUDI sampler permitted the capture of airborne mycobacteria in discrete size ranges, an important parameter for relating the airborne mycobacteria cells to potential respirable particles (aerodynamic diameter <10 microm) capable of causing health effects. Analysis of the samples was rapid, requiring only 1-1.5 days, as no microbial culturing or DNA purification was required. This approach was then used to detect suspected mycobacteria contamination associated with pools at a large public facility. PCR was also used to analyze various water samples from these pools. Again, no culturing or sample purification was required. Water samples taken from all ultraviolet light/hydrogen peroxide treated whirlpools tested positive for the presence of mycobacteria. No mycobacteria were detected in the chlorine-treated pools and the water main supply facility. All air samples collected in the proximity of the indoor whirlpools and the associated changing rooms were strongly positive for airborne mycobacteria. The airborne mycobacteria particles were predominantly collected on MOUDI stages 1-6 representing an aerodynamic size range of 0.5 to 9.9 microm. In conclusion, using this approach permits the rapid detection of mycobacteria contamination as well as the routine monitoring of suspected pools. The approach circumvents problems associated with culture-based methods such as fungal overgrowth on agar plates, and the presence of nonculturable or difficult to culture mycobacteria strains. PMID- 12650549 TI - The effectiveness of handheld ventilated sanders in reducing inhalable dust concentrations. AB - Ventilated sanders are commonly used during aircraft surface abrasion but there is limited data on their effectiveness in reducing worker exposures. This study compared two handheld ventilated sander brands, DCM and Dynabrade, in a laboratory glovebox. Both sanders collect particulates by drawing air through holes in the sanding pads; the dust subsequently passes into a vacuum collection system. Aluminum panels coated with aircraft epoxy primer and polyurethane paint were abraded and inhalable dust concentrations were measured inside the glovebox with IOM samplers. The results indicate that both sanders effectively control inhalable dust, with the DCM sander reducing mass concentrations by 93 percent, and the Dynabrade by 98 percent, when the ventilation system is used. The Dynabrade unit, however, was more aggressive and produced over four times as much dust per unit time as the DCM unit. In spite of this, the Dynabrade sander adequately collected this additional dust. Varying abrasive grit size did not significantly affect dust generation, although the differences between the grit sizes used (180 and 240 grit) were not great and may have influenced the results. PMID- 12650550 TI - Symptoms of musculoskeletal disorders among apprentice construction workers. AB - Musculoskeletal disorders (MSDs) are a major cause of work-related disability and lost-time illnesses for many occupational groups. This study determined the prevalence of musculoskeletal symptoms among young construction workers. A symptom and job factors survey was self-administered to 996 construction apprentices. Prevalence was determined by the percent of positive responses to musculoskeletal symptom questions. Odds ratios and 95 percent confidence intervals were the measures of association between prevalent musculoskeletal symptoms and demographic, leisure, and job factors and were determined by logistic regression. The low back was the site most commonly reported for job related musculoskeletal symptoms (54.4%), which was also the most common reason for seeking care from a physician (16.8%) and missing work (7.3%). Number of years worked in the construction trade was significantly associated with knee (p trend = 0.0009) and wrist/hand (p-trend < 0.04) MSD symptoms and was suggestive of an association with low back pain (p-trend = 0.05). "Working in the same position for long periods" was the job factor identified as most problematic, with 49.7 percent of all construction apprentices rating it as a moderate/major problem contributing to musculoskeletal symptoms. Musculoskeletal symptoms are a significant problem among young construction workers at the beginning of their careers. Prevention strategies are needed early in the apprentice training program to reduce the potential disability associated with work-related musculoskeletal symptom disorders. PMID- 12650551 TI - Evaluation of the rate, location, and morphology of perforations in surgical gloves worn in urological operations. AB - Surgical perforation or intrinsic defects within the polymer membranes of surgical gloves are responsible for an increased exposure, and consequently infection risk, to HIV and HCV or other infectious diseases for the medical healthcare professional. A prospective field study was undertaken to evaluate the number and location of glove perforations, tested by the waterfill test. The morphology of these perforations was then inspected by stereo light and scanning electron microscopy. By comparing in-use defects to simulated defects created by medical instruments in unused gloves it was possible to categorize perforations caused by intrinsic defects or extrinsic mechanical stress. In six different types of surgical gloves collected from 720 urological operations the waterfill test revealed leakage rates of 6.6 percent to 12.3 percent. Further assessment of these gloves revealed intrinsic polymer material defect rates of 0.2 percent to 3.3 percent. The in-use acquired instrumental and mechanical perforation rate ranged from 4.6 percent to 12.15 percent. Perforations were mostly located at the index finger (left > right) or the thumbs. The morphology of the intrinsic defects was visualized as either incorporated membranous air bubbles or granular areas. The simulation of defects by surgical instruments in unused gloves resulted in the production of a catalog for comparison with defects seen in used gloves. This knowledge of the location and cause of these perforations highlights the need for a better avoidance strategy and/or establishment of a schedule for routine glove change by surgeons. In addition, the intrinsic polymer film quality of gloves can be improved by a lower incidence of air bubbles together with less granular sites. PMID- 12650552 TI - Use of the Transtheoretical Model for individuals with co-occurring disorders. AB - Because of the interactive nature between their mental disorder and their substance-use disorder, individuals with co-occurring disorders may experience factors that complicate behavioral change, such as considering abstinence from alcohol and drugs. This study examined the relationships between the stages-of change and the processes of change, constructs of the Transtheoretical Model by Prochaska and DiClemente. There was partial support for the theory. The findings suggest that individuals with co-occurring disorders require support and clinical interventions throughout the entire change process. PMID- 12650553 TI - Direct and indirect time inputs and assertive community treatment. AB - Assertive Community Treatment (ACT) is an appealing community program model because it proposes to provide individuals with coping skills that allow them to maintain independent lives in their communities and it offers the potential to decrease inpatient stays and increase community tenure. But, it is not without its limitations. Critics point out that ACT's unique elements also make it a potentially very resource intensive program--an important consideration in times of fiscal constraints. Though the charge may be justified, there is little in the literature that actually quantifies the intensity of resources used. The process through which it achieves outcomes is not frequently described. Using ACT team workload information, we examine the time trade-offs--direct for indirect time- involved with implementing this model. In addition, we describe the specific activities that make up the direct and indirect time inputs that go into supporting clients in the community through assertive team oriented case management. PMID- 12650554 TI - Depressive symptoms and psychiatric distress in low income Asian and Latino primary care patients: prevalence and recognition. AB - The aims of the study were to: (a) assess the degree to which primary care physicians recognize psychiatric distress among an ethnically diverse primary care sample composed primarily of Asians and Hispanics; and (b) to investigate the relationship between patient and physician sociodemographic factors and overall diagnostic recognition of psychiatric distress. The study sample is comprised of 252 consecutively-selected patients and eleven primary care general internists from general medicine clinics in a large public ambulatory medical center. The measures used were the Center for Epidemiologic Studies-Depression (CES-D) scale, a demographic questionnaire, and an acculturation scale; these measures were completed during interviews conducted by trained bilingual research assistants. Physicians completed a mental health treatment summary immediately after the patient's visit. Hierarchical logistic regression analyses were performed in order to examine: (1) the degree to which providers identified psychiatric distress and (2) overall diagnostic recognition among this sample in relation to demographic characteristics and degree of acculturation. As measured by the CES-D, almost one half (47.3%) of the Latino and 41.6% ofthe Asian patients had depressive symptoms indicative of psychiatric distress. In contrast, physicians identified 43.8% of Latino patients and only 23.6% of Asian patients as being psychiatrically distressed (p < .01). Physicians were more likely to classify Latinos and those with higher acculturation status as distressed (p < .01 and p < .05, respectively). Higher patient acculturation status was the only factor significantly associated with overall diagnostic recognition (p < .05), as measured by physician agreement with the CES-D. Being Asian and/or having low acculturation levels may put the patient at risk for non-detection of psychiatric distress. The high prevalence of distress lends support to initiating improved methods for screening and detection of depression among low income and racially diverse primary care patients. PMID- 12650555 TI - Adolescent treatment outcome in a community mental health center. AB - To assess the effectiveness of a community mental health center outpatient adolescent treatment program, outcome measures were completed by the parents of 50 consecutively admitted adolescent patients who were evaluated and treated with one of three therapy modalities. Their results were compared to outcome measures of 29 non-treated adolescent control subjects. Also, self-report outcome measures were completed by 30 treatment subjects. The results demonstrated a significant positive change in the study group following treatment as compared with the control group. Each of the treatment modalities used with the study group was associated with improvement. The staffing referral mechanism used to determine the particular treatment modality for each patient was shown to be consistently and appropriately implemented. PMID- 12650556 TI - The Dawn Project: a model for responding to the needs of children with emotional and behavioral challenges and their families. AB - Systems of care represent a fundamental departure from traditional service provision by espousing genuine family-centered, culturally competent philosophies and blending the funding streams of multiple payers (e.g., education, mental health, child welfare). In Marion County, Indiana, local leaders created a system of care based on these principles called the Dawn Project. Currently, a comprehensive, multidisciplinary evaluation is being implemented to evaluate the program. Preliminary findings from initial evaluation efforts suggest that for youth in the project, there is significant clinical improvement during the first year of receiving services, a reduction in the use of more restrictive settings, and a decrease in recidivism among those who successfully complete the program. PMID- 12650558 TI - The impact of the "networked computer". PMID- 12650557 TI - From a 'state mental hospital' to new homes in the city: longitudinal research into the use of intramural facilities by long-stay care-dependent psychiatric clients in Amsterdam. AB - In 1986, the traditional psychiatric hospital where 70% of Amsterdam's intramural treatment took place, was closed down. The progress of two groups of long-stay patients was followed by longitudinal cohort research for five years after their transferral to new small-scale facilities in Amsterdam. The ADL-functioning of the most severely handicapped clients improved and their psychiatric symptoms decreased, while no improvement was seen in the functioning of the more independent and less handicapped clients. Now, thirteen years after closure of the old hospital, the use of the psychiatric facilities by these clients is being evaluated. This evaluation makes clear that the improvement of the severely disabled patients depends on the intensity of care given. Apparently, the improvement in the new facilities is no guarantee for a further development into a less care-intensive environment. The less handicapped and more independent clients, however, could more easily be transferred to less care-intensive facilities. PMID- 12650559 TI - Rationale for internet-mediated communities. AB - This article discusses the theoretical foundations of Internet-mediated communities (IMCs), i.e., groups of people who share interests and make use at some time of the same Internet tools (for instance, a web site-the special concern of this study) in order to exchange information about the shared interests. From November 1999 to February 2001 a rationale was developed and validated for such communities and their web sites, due to the lack of theory unification in the field. The authors carried out five case studies of widely known IMCs, seven action researches with established groups of people who were given customized web sites to mediate their interaction, and an exploratory research supported by open-ended interviews with those groups. Additional empirical research is now needed to apply the theoretical grounds here developed to varying IMC contexts and to different communication technologies. PMID- 12650560 TI - Viability of telework at PROCEMPA. AB - At the end of the 20th century, telework appears as one of the modalities of flexible work, which is related to new organizational structures as well as to increasing use of technology. It revolutionizes the traditional ways of performing work. Its implementation creates a number of questions to be answered by the organizations and the individuals involved. This article presents a case study on the viability of implementing telework at Procempa (The Data Processing Company of the City of Porto Alegre). The case study analyzes the technical, organizational, psychological, legal, and labor union dimensions. As a result of this study, we can identify the organization's stage of readiness for telework, the conditions under which it would be implemented, and the specific issues of an implementation. PMID- 12650561 TI - Flow in journalistic telework. AB - Since the 1980s, the strengthening and dissemination of telework has motivated studies to verify its advantages and disadvantages to enterprises and workers, specially focusing on economy, quality, and productivity performances. This paper developed from a master's thesis research which analyzed the subjective impact of telework concerning workers' personal satisfaction, their perception about suffering and pleasure when developing their work activities and social interrelationships, and, particularly, the influence it could have in their maximum experiences, the flow. Considering that the press media represents a historical telework, it was chosen as the research focus, selecting columnists from a large newspaper, located in the Brazilian south region. PMID- 12650562 TI - Interpersonal communication and creativity in journalistic telework. AB - Man and work histories are interlaced to tell us how the interaction among different human groups have helped in the development of intellectual capacities of our species. Creativity is mostly seen as a gift or an individual quality, for whose bloom and exercise there are internal and external factors, understood as stimulants of the creative process. Research for a master's thesis had, as its principal aim, the analysis of the subjective impact of telework on the workers personal satisfaction. Our second category authenticates the importance of interpersonal communication among fellow workers as incentive to personal creativity. The study was developed with columnists of a great newspaper with a big circulation in the south of Brazil, because telework is a tool of the press media, and creativity is a requirement for journalistic employment. PMID- 12650563 TI - Impacts and trends of journalistic telework: the journalists' viewpoint. AB - This article, the last of a trilogy, presents general reflections about technological advances, especially considering cyberspace as a way of information transmission and exchange, and on telework forms. Based on research for an administration masters' thesis, the derivation process that resulted in the last final analysis category illustrates the feelings of the interviewed journalists about telework effects on their professional and personal lives. PMID- 12650564 TI - Psychology and the internet: a social ecological analysis. AB - This paper proposes a research strategy based on a social ecological analysis of the Internet and its psychological impact as an option to generate original research to answer the follow ing question: What is the psycho-environmental meaning of the Internet? This paper has two objectives: first, to analyze Computer Mediated Communication (CMC) linked to the use of the Internet from a social ecological approach, and second, to propose some relationships among variables from a social ecological perspective, which can help to clarify the variability and magnitude of the psychosocial effect associated with the Internet. This article is divided into three sections. The first briefly describes the origins and development of the Internet. It identifies some technological features and specifies some of the cutting-edge breakthroughs that have facilitated its expansion. The second section proposes a conceptual scheme from the social ecological perspective, which analyzes the subject-environmental binomial associated with the Internet. It identifies the basic assumption, the conceptual richness, and possibilities for research on the Internet, using a social ecological approach. Finally, the last section describes the scope and limitations of this perspective, and discusses its heuristic utility for the development and consolidation of a new area in psychological research: "digital psychology or cyber psychology." PMID- 12650565 TI - The use of the internet in psychological research: comparison of online and offline questionnaires. AB - The Internet can be an effective medium for the posting, exchange, and collection of information in psychology-related research and data. The relative ease and inexpensiveness of creating and maintaining Web-based applications, associated with the simplicity of use via the graphic-user interface format of form-based surveys, can establish a new research frontier for the social and behavioral sciences. To explore the possible use of Internet tools in psychological research, this study compared Web-based assessment techniques with traditional paper-based methods of different measures of Internet attitudes and behaviors in an Italian sample. The collected data were analyzed to identify both differences between the two samples and in the psychometric characteristics of the questionnaires. Even if we found significant differences between the two samples in the Internet attitudes and behaviors, no relevant differences were found in the psychometric properties of the different questionnaires. This result, similar to the ones previously obtained in Web-based assessments of personality constructs, is even more interesting given the lack of control on the characteristics of the online sample. These finding suggests that, if sampling control and validity assessment is provided, Internet-based questionnaires can be a suitable alternative to more traditional paper-based measures. PMID- 12650566 TI - Breaking the stereotype: the case of online gaming. AB - Despite the rise of computer games as a leisure phenomenon, there has been relatively little research into this area. Furthermore, almost all of the research to date has concentrated on arcade or console games. More recently, the Internet has become a new medium in which players can play videogames. Since there is no published research in this area, some "benchmark" data on which future research can build was collected from two online gaming fan sites. Sociodemographics showed that the majority of players were male (approximately 85%). Over 60% of players were older than 19 years. The data provide clear evidence that the game clientele is very much an adult profile and suggest a different picture to the stereotypical image of an adolescent online gamer. The stereotype of the typical online player being a socially withdrawn young male with limited sex role identity appears to be misplaced. PMID- 12650567 TI - Model of problematic internet use in people with a sexual interest in children. AB - Agencies working with sex offenders are starting to see the emergence of people with a sexual interest in children who meet some of their needs through the use of child pornography, or the seduction of children, through the Internet. While CBT models dominate our understanding of sex offenders, there has been little research into the role that such new technologies may play in offending behavior. Data from the COPINE project has been used to generate a model of such offending behavior that emphasizes the role of cognitions in both the etiology, engagement with and problematic use of the Internet for those with a sexual interest in children. Such a model seeks to incorporate contemporary thinking about the role of cognitions in Pathological Internet Use, but applies this from a nonpathological perspective. This model is a first step towards providing a conceptual framework for such offending that will help inform both assessment and therapy. PMID- 12650568 TI - Cobalt blood levels after total hip replacement (THR): a new follow-up study in Trieste (Italy). AB - Cobalt and chromium release in patients undergone a metal-on-metal total hip replacement (THR) is a matter recently discussed and whose we do not have enough information about it yet. In literature there is little data and not completely indicative, in the biological fluids and organs the amounts of released metals are different. This is also due to the fact that cobalt and chromium blood levels can change depending on physical and working activity, individual feeding and metabolism. The results obtained confirm the presence of an increase of cobalt inthe blood of patients after total hip replacement, while the chromium levels are almost alike: average values in patients operated are 4.1 +/- 1.5 microg/L for cobalt (0.3 +/- 0.1 microg/L in the control group) and 4.5 +/- 2.9 microg/L for chromium (4.7 +/- 2.4 microg/L in the control group). In spite of the cobalt values stand below the concentration generally considered dangerous, the difference between the two examined groups points out that a risk exists for the health of these patients. These results must be confirmed by further studies, providing better information and more reliable and biocompatible materials. PMID- 12650569 TI - The chemical state of arsenic in minerals of environmental interest--an XPS and an XAES study. AB - A systematic analytical study using X-ray photoelectron spectroscopy (XPS) and X ray induced Auger electron spectroscopy (XAES) has been carried out to characterize the chemical state of arsenic in complex environmental samples. The conventional approach, which relies on the chemical shift of the core levels As3d, provides ambiguous results in determining the chemical environment of arsenic. A more accurate approach, based on the Auger parameter and on the Wagner (Chemical State) plot, which combines AsLMM kinetic energy and As3d binding energy, was adopted. This novel method for determining the chemical state of arsenic was employed to completely characterize arsenic in complex environmental samples. PMID- 12650570 TI - Description of the carbonaceous particulate matter evolution in an urban area. AB - The ground concentrations of particulate matter, total carbon, elemental carbon and organic carbon were measured in a monitoring station located inner a park in downtown Rome. The description of the evolution of the boundary layer is given by radon concentration measurements. The results show that the particulate matter of Rome contains about 30% of carbonaceous matter. The elemental and organic carbon behaviours show that elemental carbon is essentially a primary pollutant emitted directly from vehicular traffic and organic carbon behaviour instead can be primary (combustion and biogenic emission) and secondary. A chemical fine particulate matter mass balance is given; an organic carbon investigation has been carried out in order to determine n-alkanes, n-alkanoic acids and polycyclic aromatic hydrocarbons. PMID- 12650571 TI - Evaluation of indoor BTX in an outskirt zone of Rome (Italy). AB - Correlation between indoor and outdoor levels of BTX (benzene, toluene and the sum of ethyl benzene, o-xylene, m-xylene and p-xylene) has been investigated at thirteen homes, thirteen classrooms and related outdoor atmospheres. Non smoking people and low polluted sites were chosen for this campaign, in order to highlight if commonly used domestic materials could act as internal sources. Data, obtained by employing long-term diffusive samplers over a three-month period, are compared with a parallel experiment showing the day-night indoor/outdoor trend. The obtained results let us conclude that there is evidence of internal sources at homes whilst in the schools depletion phenomena prevail, probably due to the walls adsorption. PMID- 12650572 TI - Pseudomonas putida cell biosensor operating in n-hexane to determine benzene in hydrophobic matrices. AB - An immobilised Pseudomonas putida cell based biosensor, able to work directly in organic solvent (n-hexane), was designed and built. The response, in n-hexane, to benzene and some of its derivatives was studied. The proposed biosensor was found to be suitable for determining benzene in hydrophobic matrices. PMID- 12650573 TI - The measurement of atmospheric pollutants by passive sampling at the Uffizi Gallery, Florence. AB - Diffusive sampling can be adopted to analyse in detail the temporal and spatial trends of pollutants concentration in specific historic buildings or in museums. An application of this method is discussed in this paper and the results of a still ongoing monitoring campaign at the Uffizi Gallery in Florence, in the framework of an EU-founded Project (MIMIC), are presented. Distinct trends in the concentration of NO2, NOx, HONO, HNO3, O3 and SO2 have been observed in eight successive sampling periods from March 2001 to February 2002. Considerations are given on possible implications for the conservation of indoor artworks. PMID- 12650574 TI - Cluster analysis: significance, empty space, clustering tendency, non-uniformity. I--Statistical tests on the significance of clusters. AB - The agglomerative clustering methods and the tests usually applied to evaluate the significance of clusters are critically evaluated. Many clustering techniques can provide erroneous information about the existence of clusters. The single linkage technique is suggested to identify natural, well separated, clusters. The existing statistical tests on the significance of clusters are not satisfactory. A new statistical test, based on the distribution of the distances between the objects and their first nearest neighbor, is presented. The performances of the test are compared with those of the Sneath test and of the variance-ratio test on some artificial and real data sets. PMID- 12650575 TI - Study of nitrogen containing compounds in the polar troposphere. AB - Atmospheric nitrogen compounds play a critical role in tropospheric photochemistry and are crucial to understand the chemical and physical evolution of atmospheric pollutants in polar areas. Measurements of these species in remote areas are rare, although their relevance is well established. Sampling campaigns of gaseous and particulate atmospheric trace species were performed in Arctic and in Antarctica during three consecutive years (1997-1999), using a proper combination of annular denuders and filter pack. After sampling, the ionic species were extracted with aqueous solutions and analysed by means of ion chromatography. Quality assurance on the sampling and analytical steps allowed accurate and precise measurements of all relevant compounds, which are thought to be important to nitrogen chemistry, at very low concentration levels. In addition, the measurements also included a multistage low-pressure impactor for the collection of particulate matter in different size regions (0.035-15.9 mm). Results obtained from these campaigns demonstrate that the minor components may be measured at levels as low as a few nanograms per cubic meter. The reported concentrations are to be considered among the first observations of nitrogen containing compounds in polar sites. PMID- 12650576 TI - Inorganic species in water and sediments of the Lamone and Marzeno rivers ecosystem. AB - Analytical results are reported for the determination of inorganic species in water and sediments sampled in the Lamone and Marzeno rivers ecosystem. The species determined are Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Zn, Hg, F-, Cl- Br-, NO3-, SO4--, Na+, K+, Mg++, Ca++, NH4+ in superficial water, and Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Zn, Hg in sediments. For all the elements, in addition to detection limits, precision and accuracy are given: the former, expressed as relative standard deviation (s(r)), and the latter, expressed as relative error (e), were good, being in all cases lower than 6%. PMID- 12650578 TI - Review about olive oil characterization using high-field nuclear magnetic resonance. AB - In the last years Nuclear Magnetic Resonance (NMR) spectroscopy has found many applications in food analysis. Here, we present a review of the most significant results we have obtained in the olive oil characterization using NMR techniques in combination with multivariate statistical methods. PMID- 12650577 TI - A new device for reducing diesel engine emissions: a preliminary investigation. AB - A new catalytic converter is compose by a alloy noble metal supported on inert wool and located downstream to combustion chamber appears able to markedly reduce the content of toxic organics, fine particulates and CO emitted by a diesel engine equipped truck. By contrast, emissions of NOx and particulate n-alkanes remain unaltered or increase, according to the driving and operating conditions of the vehicle. Although this tesrs are preliminary the results obtained, however, suggest that the adoption of this device significantly reduce the health risk associated to fine aerosols and carcinogenic PAH. Exhaust concentrations of some unregulated species, e.g. volatile aldehydes, were lowered by the oxidising catalyzer. PMID- 12650579 TI - A new approach to the statistical treatment of 2D-maps in proteomics using fuzzy logic. AB - A new approach to the statistical treatment of 2D-maps has been developed. This method is based on the use of fuzzy logic and allows to take into consideration the typical low reproducibility of 2D-maps. In this approach the signal corresponding to the presence of proteins on the 2D-maps is substituted with probability functions, centred on the signal itself. The standard deviation of the bidimensional gaussian probability function employed to blur the signal allows to assign different uncertainties to the two electrophoretic dimensions. The effect of changing the standard deviation and the digitalisation resolution are investigated. PMID- 12650580 TI - The atmospheric monitoring in a protected area. AB - A multi-annual research program was carried out to study the environmental quality of Castelporziano Presidential Estate (Rome, Italy). Within this program, in the field of air quality, a methodological approach was defined and applied, even by means of proper Environmental Indicators for the identification of anthropogenic contribution and the quantification of degradation. By means of mobile laboratories, macro and micro-pollutant concentrations were assessed in order to define Indexes of Atmosphere Quality and Diffuse Contamination, by relating them to possible short or long-range emission sources. Wet and dry atmospheric depositions were collected and analysed for the determination of heavy metal and acid species fluxes. Critical Load and relative Exceedance maps were elaborated, for the purpose of better underline the areas characterized by a higher environmental vulnerability. PMID- 12650581 TI - Bentonite: an industrial mineral at man kind service. AB - This paper presents a general overview on the structure and chemico-physical properties of bentonites, their ion exchange properties, their main uses and applications, and the foreseen applications which may be envisaged for the next future. PMID- 12650582 TI - Transport of atmospheric pollutants in a semi-rural area of central Italy. AB - An environmental monitoring program aimed to the evaluation of the air quality in the surroundings of Gallese (near Viterbo, in central Italy) was carried out from July 1998 to July 2000. The goal of the program was the comprehension of the transport and transformation mechanisms of nitrogen and sulphur pollutants, and the evaluation of photochemical smog episodes. In this framework, a wide range of atmospheric pollution events was studied, and for many of them it was possible to detect the main source of the pollution. We report in this paper the interpretation of ozone episodes, sulphur dioxide episodes and sea-salts episodes, due to transport from the urban area of Rome, the industrial area of Narni-Tarni and the coastal areas of the Tyrrhenian Sea, respectively. PMID- 12650583 TI - Simultaneous determination of HCHO, CH3CHO and O(x) in ambient air by hydrazine reagent and hplc. AB - Formaldehyde, acetaldehyde, ozone and nitrogen dioxide in ambient air are simultaneously collected on silica gel cartridges coated with 1-methyl-1-(2,4 dinitrophenyl)hydrazine (MDNPH), where the two aldehydes are derivatized to their respective hydrazones, while the two oxidants are converted into N-methyl-2,4 dinitroaniline (MDNA). The three products are then separated and quantified by HPLC with UV detection at 360 nm. The stoichiometric factors of the MDNPH reactions with O3 and NO2 in laboratory tests correspond to 2.0 +/- 0.1 moles of MDNA per mole of O(x) (O3 + NO2). The limits of detection (LOD) are 0.7 ppbv HCHO, 0.8 ppbv CH3CHO and 1.6 ppbv O(x) for 30 L (1 h) air sampled. The sampling performance is insensitive to relative humidities encountered in real atmospheres. When compared with Sep-Pak DNPH silica cartridges as well as with ozone photometric and nitrogen dioxide chemiluminescent analyzers, the proposed chromatographic method demonstrates a very good accuracy (12% for HCHO, 14% for CH3CHO and 7% for O(x), on the average) under field sampling conditions at concentrations lower than 3 and 1 ppbv, for HCHO and CH3CHO, respectively and ranging from 28 to 62 ppbv for O(x). PMID- 12650584 TI - Microorganic pollutants in the outskirts of Rome. AB - A short field campaign was performed in the outskirts of Rome at four sites located pretty along the four rose wind directions to city centre. Both chlorinated (PCB and PCDD/F) and non-chlorinated (n-alkanes, PAH, nitrated-PAH, n alkanoic acids) organic micropollutants were investigated for their contents in the atmosphere. Concentrations reached by these pollutants in the outskirts were compared to those found in downtown Rome, both inside and outside of its largest city garden. Although concentrations of organic pollutants found in the outskirts were quite low, however they seemed enough high to induce some health risk in humans. Rural sites were less affected than industrial and waste disposal/treatment areas. PMID- 12650585 TI - Ulivo alchemy: the bio- and tecno-molecular approach to MAC-Mediterranean aliment culture. AB - Sccoiridoid biophenols, contained in olive drupes, are enzymatically activated to provide a complex mechanism for the biological defence against pathogen attack. Their identification, quantitation and metabolic behaviour are investigated, by HPLC and NMR experiments, for the improvement of olivegrowing and of the production of oil and table olives. PMID- 12650587 TI - Oil-fly ash and black crust formation in monuments. PMID- 12650586 TI - Ultra trace determination of Pt and Rh in wastewater and gullypot sediments from a low polluted area. PMID- 12650588 TI - Structure-based QSAR study on differential inhibition of human prostaglandin endoperoxide H synthase-2 (COX-2) by nonsteroidal anti-inflammatory drugs. AB - The prostaglandin-endoperoxide H synthase-1 (PGHS- 1) and prostaglandin endoperoxide H synthase-2 (PGHS-2) are the targets of nonsteroidal anti inflammatory drugs (NSAIDs). It appears that the high degree of selectivity for inhibition of PGHS-2 shown by certain compounds is the result of two mechanisms (time-dependent, time-independent inhibition), by which they interact with each isoform. Molecular models of the complexes formed by indomethacin, sulindac, fenamates, 2-phenylpropionic acids and selective cyclooxygenase-2 (COX-2) inhibitors with the cyclooxygenase active site of human PGHS-2 have been built, paying particular attention to water molecules that participate in the hydrogen bonding network at the polar active site entrance. The stability of the complexes has been assessed by molecular dynamics simulations and interaction energy decomposition analysis, and their biological significance has been discussed in light of available X-ray crystallographic and kinetic results. The selective PGHS 2 inhibitors exploit the extra space of a side-pocket in the active site of PGHS 2 that is not found in PGHS-1. The results suggest that active site hydration together with residues Tyr355, Glu524, Arg120 and Arg513 are crucial to understand the time-dependent inhibition mechanism. A marked relationship between the isoform selectivity and tightly interactions with residues into the side pocket bordered by Val523 is also found. PMID- 12650589 TI - Megavariate analysis of hierarchical QSAR data. AB - Multivariate PCA- and PLS-models involving many variables are often difficult to interpret, because plots and lists of loadings, coefficients, VIPs, etc, rapidly become messy and hard to overview. There may then be a strong temptation to eliminate variables to obtain a smaller data set. Such a reduction of variables, however, often removes information and makes the modelling efforts less reliable. Model interpretation may be misleading and predictive power may deteriorate. A better alternative is usually to partition the variables into blocks of logically related variables and apply hierarchical data analysis. Such blocked data may be analyzed by PCA and PLS. This modelling forms the base-level of the hierarchical modelling set-up. On the base-level in-depth information is extracted for the different blocks. The score vectors formed on the base-level, here called 'super variables', may be linked together in new matrices on the top-level. On the top level superficial relationships between the X- and the Y-data are investigated. In this paper the basic principles of hierarchical modelling by means of PCA and PLS are reviewed. One objective of the paper is to disseminate this concept to a broader QSAR audience. The hierarchical methods are used to analyze a set of 10 haloalkanes for which K = 30 chemical descriptors and M = 255 biological responses have been gathered. Due to the complexity of the biological data, they are sub-divided in four blocks. All the modelling steps on the base-level and the top-level are reported and the final QSAR model is interpreted thoroughly. PMID- 12650590 TI - Combining NMR spectral and structural data to form models of polychlorinated dibenzodioxins, dibenzofurans, and biphenyls binding to the AhR. AB - A three-dimensional quantitative spectrometric data-activity relationship (3D QSDAR) modeling technique which uses NMR spectral and structural information that is combined in a 3D-connectivity matrix has been developed. A 3D-connectivity matrix was built by displaying all possible assigned carbon NMR chemical shifts, carbon-to-carbon connections, and distances between the carbons. Two-dimensional 13C-13C COSY and 2D slices from the distance dimension of the 3D-connectivity matrix were used to produce a relationship among the 2D spectral patterns for polychlorinated dibenzofurans, dibenzodioxins, and biphenyls (PCDFs, PCDDs, and PCBs respectively) binding to the aryl hydrocarbon receptor (AhR). We refer to this technique as comparative structural connectivity spectral analysis (CoSCoSA) modeling. All CoSCoSA models were developed using forward multiple linear regression analysis of the predicted 13C NMR structure-connectivity spectral bins. A CoSCoSA model for 26 PCDFs had an explained variance (r2) of 0.93 and an average leave-four-out cross-validated variance (q(2)4) of 0.89. A CoSCoSA model for 14 PCDDs produced an r2 of 0.90 and an average leave-two-out cross-validated variance (q(2)2) of 0.79. One CoSCoSA model for 12 PCBs gave an r2 of 0.91 and an average q(2)2 of 0.80. Another CoSCoSA model for all 52 compounds had an r2 of 0.85 and an average q(2)2 of 0.52. Major benefits of CoSCoSA modeling include ease of development since the technique does not use molecular docking routines. PMID- 12650591 TI - The consequences of translational and rotational entropy lost by small molecules on binding to proteins. AB - When a small molecule binds to a protein, it loses a significant amount of rigid body translational and rotational entropy. Estimates of the associated energy barrier vary widely in the literature yet accurate estimates are important in the interpretation of results from fragment-based drug discovery techniques. This paper describes an analysis that allows the estimation of the rigid body entropy barrier from the increase in binding affinities that results when two fragments of known affinity and known binding mode are joined together. The paper reviews the relatively rare number of examples where good quality data is available. From the analysis of this data, we estimate that the barrier to binding, due to the loss of rigid-body entropy, is 15-20 kJ/mol, i.e. around 3 orders of magnitude in affinity at 298 K. This large barrier explains why it is comparatively rare to observe multiple fragments binding to non-overlapping adjacent sites in enzymes. The barrier is also consistent with medicinal chemistry experience where small changes in the critical binding regions of ligands are often poorly tolerated by enzymes. PMID- 12650592 TI - Analysis methods for identifying coordinated movements during ligand unbinding. AB - Molecular dynamics simulations have been applied to unbind biological ligands from their receptors. Conformation changes are observed in the biomolecules during unbinding, but there exists no systematic method to detect these conformation changes. In this work, we have used 'essential dynamics' (ED) and projection to latent structures (PLS) to investigate the conformation changes of the bovine serum retinol-binding protein when retinol unbinds from its receptor site. The results of these analyses characterise a large proportion of the movements that occur during unbinding. We find that the loop regions of retinol binding protein exhibit the largest movements during unbinding. The sudden changes in unbinding speed during the unbinding process appear not to be caused by sudden changes in protein structure. PMID- 12650593 TI - Allergenic evaluation of Malassezia furfur crude extracts. AB - Crude extracts of the lipophilic yeast Malassezia furfur were obtained from 2, 6, 10 and 28 day old cultures. The in vitro cultivation periods corresponded, respectively, to the lag phase, middle of the log phase, end of log phase and the decline phase of the growth curve, which was based on viable cell counts obtained with a fluorescent viability test. Biochemical analyses showed that the protein and carbohydrate contents were greater in day 10 extracts. Seventy patients with different allergic manifestations and 30 healthy volunteers were skin prick tested using the extracts. Of these, thirteen (18.57%) patients gave positive responses. SDS PAGE gradient electrophoretic profiles of the preparations indicated that the 28 day extracts contained the greatest number of protein bands with molecular weights ranging mostly between 30 and 94 kDa. Immunoblots incubated with individual patient sera showed that four IgE binding M. furfur allergens of approximately 88, 61, 52 and 39 kDa were present in the 28 day extracts. The components identified could be used for detecting IgE mediated responses to M. furfur among individuals affected with different allergic conditions. PMID- 12650594 TI - Experimental reproduction of the Jorge Lobo's disease in BAlb/c mice inoculated with Lacazia loboi obtained from a previously infected mouse. AB - Long-term maintenance of Lacazia loboi in the laboratory has not been reported. We report here the use BALB/c mice to maintain the Lacazia loboi for extended period of time. Eight to ten week-old mice were inoculated intradermally in both hind footpads with a fungal suspension from a macerated footpad obtained from an original mouse previously infected with the fungi and sacrificed 8 months after inoculation. The inoculated animals were sacrificed at different time intervals, footpads were excised, the right one was submitted to histopathological examination and the left one was macerated in sterile saline for fungal count and viability index determination. The inoculated animals presented the histopathological picture identical to the mice previously inoculated with material from human lesion. Granulomatous infiltrates with predominance of macrophages and giant cells were observed. The granulomas evolved progressively as observed in the different times of sacrifice. After 7 months of inoculation, macroscopic lesions were observed, and the number of fungi obtained from macerated footpads was higher than the number of inoculated fungi. The pattern of lesion development was similar to what was observed in animals infected with a fungal suspension obtained from a human lesion. Considering the histopathological findings, the clinical manifestations, and the finding of a higher number of fungi obtained than the inoculated into footpads of each mice, we believe the BALB/c mice strain is as an excellent way to maintain L. loboi in laboratory. Moreover, even after serial passages of the fungi, the granulomatous lesions are reproduced consistently in laboratory conditions. PMID- 12650595 TI - The fungicidal activity of novel nanoemulsion (X8W60PC) against clinically important yeast and filamentous fungi. AB - Surfactant nanoemulsions are water in oil preparations that proved to have a broad spectrum biocidal activity against a variety of microorganisms including Gram-positive and Gram-negative bacteria, spores and enveloped viruses. These preparations are non-toxic to the skin, mucous membrane and gastrointestinal tissues at biocidal concentrations. In this study, 0.1% of the nanoemulsion designated X8W60PC has shown fungicidal activity against yeast including Candida albicans and C. tropicalis in 15 minutes. C. tropicalis was more sensitive than C. albicans, which required a longer time or a higher concentration of the nanoemulsion to achieve killing. Neutral to slightly alkaline pH was more effective in killing the yeast cells than acidic pH. Using the minimum inhibitory concentration assay, 0.08% of the nanoemulsion was inhibitory to C. albicans, and parapsilosis and filamentous fungi including Microsporum gypseum, Trichophyton mentagrophytes, Trichophyton rubrum, Aspergillus fumigatus and Fusarium oxysporum. None of the individual ingredients was as effective a fungicidal as the nanoemulsion at equivalent concentration. This shows that the nanoemulsion structure is an important factor in the anti-fungal activity. The X8W60PC has great potential as a topical anti-fungal agent and further investigation into the mechanism of fungicidal action is warranted. PMID- 12650596 TI - Dermatophytes isolated from patients in University Hospital, Kuala Lumpur, Malaysia. AB - A total of 576 dermatophytes were isolated from patients with a variety of skin infections from January 1993 to May 2000. Ten species of dermatophytes were identified: Epidermophyton floccosum (0.7%), Microsporum audouinii (1.1%), M. canis (3.1%), M. gypseum (0.3%), Trichophyton concentricum (3.5%), T. equinum (0.2%), T. mentagrophytes (36.1%), T. rubrum (53.8%), T. verrucosum (0.2) and T. violaceum (1.0%). The body sites most frequently affected by dermatophytes were the buttocks, nails and trunk. Anthropophilic dermatophytes made up 60.1% of the isolates; the most common species was T. rubrum, T. mentagrophytes and M. canis were the two main zoophilic dermatophytes. T. mentagrophytes was isolated from all body sites except the scalp. M. canis was found to be associated with domestic dogs and was not isolated from ethnic Malays. The only geophilic dermatophyte was M. gypseum, an uncommon dermatophyte associated with tinea pedis. PMID- 12650597 TI - Mycoflora of freshly harvested flint corn from Northwestern Provinces in Argentina. AB - A mycological survey was carried out for the first time, on red flint corn samples from the northwestern Andinian region of Argentina in the 1999 and 2000 harvest seasons. Species of the genus Fusarium were the most prevalent component of the flint corn mycoflora present in all provinces. F. verticillioides was the predominant Fusarium isolated in the 1999 harvest season in the the region, and was found at higher incidence level than those observed on commercial semident corn hybrids harvested in the main corn production area in Argentina (Pampean region). During the 2000 harvest season, Fusarium graminearum was most commonly isolated species in Salta province. PMID- 12650598 TI - Antifungal activity of substituted 8-quinolinol-5- and 7-sulfonic acids: a mechanism of action is suggested based on intramolecular synergism. AB - 8-Quinolinol-5-sulfonic acid was nearly devoid of antimicrobial activity, due to what was believed to be an unfavorable partition coefficient. Since twenty six 8 quinolinol-5- and 7-sulfonic acids were available from our previous work, they were tested against six fungi. The 7-chloro and 7-bromo-5-sulfonic acids and the 5-chloro and 5-bromo-7-sulfonic acids showed fungal inhibition within one order of magnitude of that of 8-quinolinol. It is suggested that a nonchelating mechanism is in part responsible for this fungitoxicity. Five additional 5 sulfonic acids with chlorine in positions 3-, 6-, 3,6-, 3,7-, and 6,7- that were suitable for studies in synergism became available more recently. The enhanced activities of the dichlorosulfonic acids over the correspondingly substituted monochlorosulfonic acids is attributed to intramolecular synergism. PMID- 12650599 TI - Accumulation of fumonisins B1 and B2 in freshly harvested Brazilian commercial maize at three locations during two nonconsecutive seasons. AB - Fifty-six Brazilian commercial maize cultivars were examined for FB1 and FB2 accumulation after two non-consecutive growing seasons. During the 94/95 growing season 35 cultivars were planted at three locations in the state of Sao Paulo, Brazil. All samples (total of 105) were contaminated (0.10 micro/g-6.58 microg/g FB1 and 0.04 microg/g-2.15 microg/g FB2). During the 97/98 growing season, 8 of the cultivars used during 94/95 and 21 others were replanted at the same locations. All 87 samples were contaminated (1.15 microg/g-43.80 microg/g FB1 and 0.08 microg/g-11.65 microg/g FB2). One cultivar accumulated significantly less fumonisins in all locations during both growing seasons, indicating that some degree of selection may be possible even in climates that favor F. moniliforme (verticillioides) infection of maize. The presence of water surplus in soil from kernel maturity to harvest correlated with concentrations of FB1 in the grain for the 8 cultivars planted during both seasons at three locations. Observed trends indicated that water excesses and deficits from silking to harvest increased fumonisin levels. The difference in the incidence of FB1, FB2, and FB1 + FB2 was significant between growing seasons, planting locations and between cultivars. Neither the level of hybridization, nor the type of endosperm, nor the length of the vegetative cycle showed any effect on the FB1 contamination. PMID- 12650600 TI - Prospects for biological control of the western flower thrips, Frankliniella occidentalis, with the entomopathogenic fungus, Metarhizium anisopliae, on chrysanthemum. AB - The potential of Metarhizium anisopliae (Metsch.) Sorok. for the control of Frankliniella occidentalis (Pergande) on chrysanthemum cuttings was evaluated in greenhouse experiments. The fungus significantly reduced both the adult and larval populations of F. occidentalis, although the level of control of larval populations was much lower than for adults. Combined application of M. anisopliae and Methomyl (Lannate), however, resulted in a significant reduction of both the larval and adult stages. The use of both control agents might be helpful in reducing the selection pressure for resistance to chemical insecticides, thereby delaying or preventing the build-up of resistant populations in greenhouses. PMID- 12650601 TI - Inhibition of signal transduction by the nm23 metastasis suppressor: possible mechanisms. AB - The first metastasis suppressor gene identified was nm23. Transfection of nm23 into metastatic cell lines resulted in the inhibition of metastasis, but not primary tumor size in vivo. Using in vitro assays, nm23 overexpression resulted in reduced anchorage-independent colonization in response to TGF-beta, reduced invasion and motility in response to multiple factors, and increased differentiation. We hypothesize that the mechanism of action of Nm23 in metastasis suppression involves diminished signal transduction downstream of a particular receptor. Candidate biochemical mechanisms are identified and discussed herein. PMID- 12650602 TI - KISS1 metastasis suppression and emergent pathways. AB - Metastatic disease is the most critical impediment to cancer patient survival. However, comparatively little is known concerning the intricate pathways which govern the complex phenotypes associated with metastasis. The KISS1 metastasis suppressor gene inhibits metastasis in both in vivo melanoma and breast carcinoma models. Despite its clear physiological activity, the mechanism of KISS1 remains unclear. Recent identification of a 54 amino acid peptide of KISS1, termed metastin or kisspeptin-54, and its cognate G-protein coupled receptor (hOT7T175, AXOR12, GPR54) have provided additional clues and avenues of research. While studies have attributed KISS1 with modulation of NFkappaB regulation, experiments with metastin and its receptor implicate MAP kinase pathways and also suggest the potential of autocrine, paracrine and endocrine roles. Impacts on motility, chemotaxis, adhesion and invasion have each been documented in disparate cell lines and conflicting observations require resolution. Nevertheless, mounting clinical evidence, particularly the loss of KISS1 in metastases, correlates KISS1 and metastin receptor expression with human tumor progression. Together, the data substantiate roles for these molecules in metastasis regulation. PMID- 12650603 TI - Tumor metastasis-associated human MTA1 gene and its MTA1 protein product: role in epithelial cancer cell invasion, proliferation and nuclear regulation. AB - Using differential cDNA library screening techniques based on metastatic and nonmetastatic rat mammary adenocarcinoma cell lines, we previously cloned and sequenced the metastasis-associated gene mta1. Using homology to the rat mta1 gene, we cloned the human MTA1 gene and found it to be over-expressed in a variety of human cell lines (breast, ovarian, lung, gastric and colorectal cancer but not melanoma or sarcoma) and cancerous tissues (breast, esophageal, colorectal, gastric and pancreatic cancer). We found a close similarity between the human MTA1 and rat mta1 genes (88% and 96% identities of the nucleotide and predicted amino acid sequences, respectively). Both genes encode novel proteins that contain a proline rich region (SH3-binding motif), a putative zinc finger motif, a leucine zipper motif and 5 copies of the SPXX motif found in gene regulatory proteins. Using Southern blot analysis the MTA1 gene was highly conserved, and using Northern blot analysis MTA1 transcripts were found in virtually all human cell lines (melanoma, breast, cervix and ovarian carcinoma cells and normal breast epithelial cells). However, the expression level of the MTA1 gene in normal breast epithelial cells was approximately 50% of that found in rapidly growing adenocarcinoma and atypical epithelial cell lines. Experimental inhibition of MTA1 protein expression using antisense phosphorothioate oligonucleotides resulted in inhibition of growth and invasion of human MDA-MB-231 breast cancer cells with relatively high MTA1 expression. Furthermore, the MTA1 protein was localized in the nuclei of cells transfected with a mammalian expression vector containing a full-length MTA1 gene. Although some MTA1 protein was found in the cytoplasm, the vast majority of MTA1 protein was localized in the nucleus. Examination of recombinate MTA1 and related MTA2 proteins suggests that MTA1 protein is a histone deacetylase. It also appears to behave like a GATA-element transcription factor, since transfection of a GATA element reporter into MTA1-expressing cells resulted in 10-20-fold increase in reporter expression over poorly MTA1-expressing cells. Since it was reported that nucleosome remodeling histone deacetylase complex (NuRD complex) involved in chromatin remodeling contains MTA1 protein and a MTA1-related protein (MTA2), we examined NuRD complexes for the presence of MTA1 protein and found an association of this protein with histone deacetylase. The results suggest that the MTA1 protein may serve multiple functions in cellular signaling, chromosome remodeling and transcription processes that are important in the progression, invasion and growth of metastatic epithelial cells. PMID- 12650604 TI - MKK4 and metastasis suppression: a marriage of signal transduction and metastasis research. AB - MAP kinase kinase 4 (MKK4) is a member of the stress-activated protein kinase (SAPK) signaling cascade and is involved in the regulation of many cellular processes. We have recently demonstrated a functional role for MKK4 in the suppression of metastases. In this review, we discuss the established cellular and biochemical functions of MKK4, as well as a new function for MKK4 as a metastasis suppressor gene. Because of the importance of signaling studies to this translational work, a detailed example of the strategy and tools that can be employed to define the biochemical mechanism of MKK4-mediated metastasis suppression is presented. Finally, the potential therapeutic utility of these findings is discussed. PMID- 12650606 TI - Breast cancer metastasis suppressor 1: update. AB - This article reviews information related to the BRMS1 (BReast Metastasis Suppressor 1) metastasis suppressor gene. BRMS1 was identified by differential display comparing metastasis-suppressed chromosome 11 hybrids with metastatic, parental MDA-MB-435 human breast carcinoma cells. BRMS1 has subsequently been shown to suppress metastasis, but not tumorigenicity of human melanoma cells. The murine version, Brms1, also suppresses metastasis and exhibits a high level of homology to the human gene at the structure, nucleotide and amino acid levels. The mechanisms of action remain to be determined; however, BRMS1 transfectant cells have restored gap junctional intercellular communication. Recent data suggest that BRMS1 is part of the mSin3A histone deacetylase complex. PMID- 12650607 TI - Overexpression of autocrine motility factor receptor (AMFR) in NIH3T3 fibroblasts induces cell transformation. AB - Autocrine motility factor receptor (AMFR) is a cell surface glycoprotein of 78000 molecular weight (gp78), regulating cell motility signaling in vitro and metastasis in vivo. To test whether AMFR could be a common mediator of transformation and oncogenic itself, we transfected NIH3T3 fibroblast cells with expression vectors carrying the full-length cDNA for mouse AMFR and evaluated the effects of increased AMFR on transforming potential. The cells stably expressing high levels of AMFR as a result of transfection displayed a complete morphological change and acquired the ability to grow even in low serum. Furthermore, they were anchorage-independent for growth in soft agar and more motile in phagokinetic track assay. Interestingly, the enhanced expression of AMFR produced tumors in nude mice. Our findings provide a direct evidence that overexpression of the AMFR is associated with the acquisition of a transformation phenotype. PMID- 12650605 TI - Regulation of cancer metastasis by stress pathways. AB - The presence of activated oncogenes and/or inactivated tumor suppressor genes may result in constitutive activation of multiple transcription factors. This may be especially true in the early stages of tumor development. At advanced stages, however, uncontrolled tumor growth and the consequent development of a stress microenvironment, such as hypoxia, acidosis, and free radical overproduction, may further alter the activity of these transcription factors. Abnormal activation of and interplay between these factors lead to aberrant expression of multiple metastasis-related proteins and confer a tremendous survival and growth advantage to emerging metastatic variants. Understanding the expression and regulation of these molecules may shed more light on the biology of cancer metastasis as well as suggest new preventive and therapeutic approaches. PMID- 12650608 TI - Overexpression and functional alterations of the EphA2 tyrosine kinase in cancer. AB - Cancer is a disease of aberrant signal transduction. The expression and function of intracellular signaling pathways are frequently subverted as cells progress towards a metastatic phenotype. In particular, tyrosine kinases initiate powerful signals that govern many different aspects of cell behavior. In Recent studies have demonstrated that the EphA2 receptor tyrosine kinase is frequently overexpressed and functionally altered in aggressive tumor cells, and that these changes promote metastatic character. Herein, we provide an overview of our current understanding of EphA2, with emphasis upon the differential regulation of EphA2 expression and function. We also show that differential EphA2 expression and function may provide a unique opportunity for selective therapeutic targeting of EphA2 in metastatic disease. PMID- 12650609 TI - Collapsin response mediator protein-1: a novel invasion-suppressor gene. AB - Numerous genetic changes are associated with metastasis of cancer cells. Previously, we used microarray to identify that collapsin response mediator protein-1 (CRMP-1) was involved in cancer invasion and metastasis. We further characterized that CRMP-1 was a novel invasion-suppression gene. Members of the CRMP gene family are intracellular phosphoproteins involved in the mediation of semaphorin induced F-actin depolymerization and growth cone collapse. The precise mechanism by which CRMP-I inhibits invasion is not yet clear. However, CRMP-1 transfected cells had fewer filopodia and less Matrigel-invasion abilities. A low expression of CRMP-I mRNA in lung cancer tissue was significantly associated with advanced disease, lymph node metastasis, early post-operative relapse, and shorter survival. In this article, we reviewed the functions of CRMPs and semaphorins and analyzed the structure and motifs of CRMP-1 by bioinformatics. As such, we hoped to shed further light on the mechanism by which CRMP-1 suppresses the invasion of cancer cells. PMID- 12650610 TI - Transcriptional regulation of osteopontin and the metastatic phenotype: evidence for a Ras-activated enhancer in the human OPN promoter. AB - Elevated osteopontin (OPN) transcription often correlates with increased metastatic potential of transformed cells, and in several model systems OPN- whether produced by the tumor cells or by stromal cells - has been shown to enhance metastatic ability. Sequence elements in the OPN promoter have been identified on the basis of their ability to interact with protein factors associated with the tumorigenic process in one or more cell lineages. One of these is a Ras-activated enhancer (RAE) that binds a protein, the Ras-response factor (RRF), whose ability to form a complex with the RAE is stimulated by Ras signaling in fibroblasts and epithelial cells. Another is the T cell factor-4 binding site, which in the OPN promoter can retard OPN transcription when bound by the Tcf-4 protein. In Rama 37 rat mammary epithelial cells Tcf-4 suppresses OPN transcription and the metastatic phenotype. A third promoter segment consists of two sequences in the -94 to -24 region of the human OPN promoter able to bind several known transcription factors, including Sp1, Myc and Oct-1, which may act synergistically to stimulate OPN transcription in malignant astrocytic cells. Although expression of other genes may also be regulated by these transcription factors, evidence suggests that often OPN alone can stimulate metastasis. In this communication we address two issues: (1) How does OPN facilitate the metastatic phenotype? (2) What mechanisms are responsible for the increase in OPN transcription in metastatic cells? PMID- 12650611 TI - Osteopontin and colon cancer progression. AB - Human colon cancer affects nearly 150,000 patients and results in 60,000 deaths in the United States per year. Despite significant advances in the management of the colon cancer patient, little change in survival rates has been appreciated over the past 50 years. The primary cause of death relates to the development of distant metastases to organs such as the liver and lungs. Colon cancer represents an important disease to study in order to better understand tumor progression and metastasis primarily because there is almost a stepwise advancement of the disease that is marked by measurable genetic and associated phenotypic alterations. Metastasis appears to be the end product of the development of 'Herculean' cell clones capable of independent growth, invasion, adhesion, avoidance of apoptosis, and angiogenesis. Although significant progress has been made in understanding the sequential genetic events leading to the development of cancer, the precise genes and the associated molecular pathways underlying the development of metastatic potential are still poorly understood. Moreover, our enhanced genetic knowledge has had relatively little trickle down effect on our clinical management of this deadly disease. For this reason, we undertook a comprehensive study to develop a molecular encyclopedia of new tumor markers and markers of tumor progression, some of which will hopefully prove useful in the clinical management of colon cancer patients by means of their capacity to detect and predict the stage and disease burden. This review will focus on the application of gene expression profiling technology to the problem of identifying new tumor markers and progression markers, and the discovery of osteopontin as the leading candidate clinical marker derived from a screen of approximately 12,000 named genes. PMID- 12650612 TI - The nectary-specific pattern of expression of the tobacco Nectarin I promoter is regulated by multiple promoter elements. AB - The major protein secreted into the nectar of tobacco plants (Nectarin I) is a germin-like protein that has superoxide dismutase activity. We have isolated the gene encoding Nectarin I (NECI) and analyzed the expression of a chloramphenicol acetyl transferase (CAT) marker gene driven by its promoter in transgenic plants. Transgenic plant lines that expressed the CAT gene under control of the full length NECI promoter showed high levels of CAT expression in mature floral nectaries. Tissue specificity of NECI-CAT expression demonstrated that the construct was expressed uniquely in nectaries with a small level of expression in ovary. Further, analysis of its temporal expression showed that the construct is expressed uniquely during those times when nectar is actively being secreted from flowers. An examination of the transcription start site verified that the initiation site of the NECI-CAT mRNA in transgenic plants is identical with that of the native gene in vivo. Two promoter deletion constructs were also prepared and analyzed. Analysis in transgenic plants revealed that the nectary-specific expression is the result of multiple promoter elements and suggests that nectar secretion and flower opening may be coordinately regulated. PMID- 12650613 TI - Neutralizing immunogenicity of transgenic carrot (Daucus carota L.)-derived measles virus hemagglutinin. AB - Although edible vaccines seem to be feasible, antigens of human pathogens have mostly been expressed in plants that are not attractive for human consumption (such as potatoes) unless they are cooked. Boiling may reduce the immunogenicity of many antigens. More recently, the technology to transform fruit and vegetable plants have become perfected. We transformed carrot plants with Agrobacterium tumefaciens to generate plants (which can be eaten raw) transgenic for an immunodominant antigen of the measles virus, a major pathogen in man. The hemagglutinin (H) glycoprotein is the principle target of neutralizing and protective antibodies against measles. Copy numbers of the H transgene were verified by Southern blot and specific transcription was confirmed by RT-PCR. The H protein was detected by western blot in the membrane fraction of transformed carrot plants. The recombinant protein seemed to have a 8% lower molecular weight than the viral protein. Although this suggests a different glycosylation pattern, proper folding of the transgenic protein was confirmed by conformational dependent monoclonal antibodies. Immunization of mice with leaf or root extracts induced high titres of IgG1 and IgG2a antibodies that cross-reacted strongly with the measles virus and neutralized the virus in vitro. These results demonstrate that transgenic carrot plants can be used as an efficient expression system to produce highly immunogenic viral antigens. Our study may pave the way towards an edible vaccine against measles which could be complementary to the current live attenuated vaccine. PMID- 12650614 TI - Molecular characterization of a novel senescence-associated gene SPA15 induced during leaf senescence in sweet potato. AB - The structure and expression of a novel senescence-associated gene (SPA15) of sweet potato were characterized. The protein coding region of the gene consists of 13 exons encoding 420 amino acids. Apparent homologues of this sweet potato gene are found in a variety of dicot and monocot plants, but not in animals or microorganisms. Examination of the expression patterns of the SPA15 gene in sweet potato reveals that the transcripts of SPA15 are specifically induced in the senescing leaves, and the temporal profile of SPA15 protein accumulation is correlated with that of SPA15 transcripts. Studies on the distribution of SPA15 homologue in rice plants also indicate that SPA15 homologue is up-regulated specifically in senescing rice leaves. Treatment of detached sweet potato leaves with phytohormones including ethylene, methyl jasmonate, salicylic acid and abscisic acid resulted in a high-level induction of SPA15. Immunoelectron microscopic analysis demonstrates that SPA15 is specifically associated with the cell wall. The potential role for SPA15 during leaf senescence is discussed. PMID- 12650615 TI - Membrane localization of Arabidopsis acyl-CoA binding protein ACBP2. AB - Cytosolic acyl-CoA binding proteins bind long-chain acyl-CoAs and act as intracellular acyl-CoA transporters and pool formers. Recently, we have characterized Arabidopsis thaliana cDNAs encoding novel forms of ACBP, designated ACBP1 and ACBP2, that contain a hydrophobic domain at the N-terminus and show conservation at the acyl-CoA binding domain to cytosolic ACBPs. We have previously demonstrated that ACBP1 is membrane-associated in Arabidopsis. Here, western blot analysis of anti-ACBP2 antibodies on A. thaliana protein showed that ACBP2 is located in the microsome-containing membrane fraction and in the subcellular fraction containing large particles (mitochondria, chloroplasts and peroxisomes), resembling the subcellular localization of ACBP1. To further investigate the subcellular localization of ACBP2, we fused ACBP2 translationally in-frame to GFP. By means of particle gene bombardment, ACBP2-GFP and ACBP1-GFP fusion proteins were observed transiently expressed at the plasma membrane and at the endoplasmic reticulum in onion epidermal cells. GFP fusions with deletion derivatives of ACBPI or ACBP2 lacking the transmembrane domain were impaired in membrane targeting. Our investigations also showed that when the transmembrane domain of ACBP1 or that of ACBP2 was fused with GFP, the fusion protein was targeted to the plasma membrane, thereby establishing their role in membrane targeting. The localization of ACBP1-GFP is consistent with our previous observations using immunoelectron microscopy whereby ACBPI was localized to the plasma membrane and vesicles. We conclude that ACBP2, like ACBP1, is a membrane protein that likely functions in membrane-associated acyl-CoA transfer/metabolism. PMID- 12650616 TI - Expression and import of an active cellulase from a thermophilic bacterium into the chloroplast both in vitro and in vivo. AB - A bacterial thermostable cellulase, the endo-1,4-beta-D-glucanase E1 from Acidothermus cellulolyticus, was imported into chloroplasts, and an active enzyme was recovered both in vitro and in vivo. Precursor fusion proteins were synthesized with E1 or its catalytic domain, CD, fused to the transit peptide of ferredoxin or ribulose-bisphosphate carboxylase activase for stromal targeting. A spacer region of 1, 5 or 15 amino acids was included carboxy to the transit peptide. The efficiency of import and processing by the stromal processing peptidase depended on the nature of the transit peptide and the passenger protein, and increased with the length of the spacer between them. Besides finding E1 or CD in the stroma, protein was arrested in the envelope during import showing that structural features of E1 and CD, along with their proximity to the transit peptide, influence translocation. The cellulose binding domain and/or serine/proline/threoline-rich linker of E1 may impede efficient import. Significantly, most precursors for E1 and CD synthesized by in vitro translation possessed endoglucanse activity that was temperature-dependent, and required the residues AGGGY at the N-terminus of E1 and CD. Furthermore, activity was detected upon import into chloroplasts. Based on the in vitro analyses, five precursor fusion proteins were selected to determine if E1 and CD would be successfully targeted to chloroplasts in vivo. In transgenic tobacco plants, E1 and CD accumulated in both the stromal and membrane fractions and, importantly, chloroplast extracts showed endoglucanase activity. PMID- 12650617 TI - Nuclease activities and DNA fragmentation during programmed cell death of megagametophyte cells of white spruce (Picea glauca) seeds. AB - The haploid megagametophyte of white spruce (Picea glauca) seeds undergoes programmed cell death (PCD) during post-germinative seedling growth. Death of the megagametophyte storage parenchyma cells was preceded by reserve mobilization and vacuolation. TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling)-positive nuclei indicated that the first megagametophyte cells to die were those closest to the radicle at the micropylar end of the seed as well as those that comprised the most peripheral and innermost layers at the chalazal end of the seed. The death process was accompanied by nuclear fragmentation and internucleosomal DNA cleavage and the sequential activation of several nucleases. The latter comprised at least two groups: those induced relatively early during post-germinative seedling growth, that had pH optima in the neutral range (33, 31, 17 and 15 kDa), and those induced later that had pH optima in the acidic range (73, 62, 48, 43 and 29 kDa). Activities of all of the nucleases were stimulated by Ca2+, Mg2+ and Mn2+; only the nucleases active at neutral pH were inhibited by Zn2+. The temporal pattern of induction of the neutral and acidic nucleases may suggest that the latter function after tonoplast rupture. PMID- 12650618 TI - Differences in the processing of DNA ends in Arabidopsis thaliana and tobacco: possible implications for genome evolution. AB - Surprising species-specific differences in non-homologous end-joining (NHEJ) of genomic double-strand breaks (DSBs) have been reported for the two dicotyledonous plants Arabidopsis thaliana and Nicotiana tabacum. In Arabidopsis deletions were, on average, larger than in tobacco and not associated with insertions. To establish the molecular basis of the phenomenon we analysed the fate of free DNA ends in both plant species by biolistic transformation of leaf tissue with linearized plasmid molecules. Southern blotting indicated that, irrespective of the nature of the ends (blunt, 5' or 3' overhangs), linearized full-length DNA molecules were, on average, more stable in tobacco than in Arabidopsis. The relative expression of a beta-glucuronidase gene encoded by the plasmid was similar in both plant species when the break was distant from the marker gene. However, if a DSB was introduced between the promoter and the open reading frame of the marker, transient expression was halved in Arabidopsis as compared to tobacco. These results indicate that free DNA ends are more stable in tobacco than in Arabidopsis, either due to lower DNA exonuclease activity or due to a better protection of DNA break ends or both. Exonucleolytic degradation of DNA ends might be a driving force in the evolution of genome size as the Arabidopsis genome is more than twenty times smaller than the tobacco genome. PMID- 12650619 TI - Centromere-specific acetylation of histone H4 in barley detected through three dimensional microscopy. AB - Histone acetylation affects chromatin conformation and transcriptional activity. However, the structural role of histone acetylation at specific chromosomal regions, such as the centromere, is poorly understood. In this study, histone H4 acetylation and its localization in barley interphase nuclei are revealed by three-dimensional microscopy. The centromeres form a ring-like allocation near the nuclear membrane in barley. Immunofluorescence studies on non-fixed, interphase nuclei treatment revealed ring-like distribution of the highly acetylated histone H4, located near the nuclear membrane at one pole of the nucleus. This fluorescent structure was similar to the centromere cluster and referred to as hyperacetylated region (HAR). The distribution pattern of the acetylated histone H4 was similar to each of the K5, K8, K12 and K16 lysine residues, although H4 acetylated at K5, K8 and K12 residues was found in almost all nuclei, whereas H4 acetylated at K16 was weakly observed in only half of the nuclei. Each HAR consists of two strongly acetylated cores and a halo-like, less acetylated surrounding area. Fluorescence signals from centromere-specific repetitive sequences of barley, detected through three-dimensional fluorescence in situ hybridization (3D-FISH), co-localized with the HAR corresponding to the K5 residue acetylation, but the signals did not completely overlap each other. These findings indicate that histone acetylation specifically occurring at the centromeres likely have certain structural roles for the centromere. PMID- 12650620 TI - Molecular characterization of rice hsp101: complementation of yeast hsp104 mutation by disaggregation of protein granules and differential expression in indica and japonica rice types. AB - HSP100 protein is an important component of the heat-shock response in diverse organisms. Using specific primers based on cDNA sequence, rice hsp101 gene was PCR-amplified and sequenced. Southern analysis revealed that there appears to be a single gene per haploid genome coding for HSP101 protein in rice. Northern analysis showed that expression of hsp101 transcript is strictly heat-inducible and induction is transient in nature. In the temperature regime tested, 45 degrees C treatment to intact rice seedlings for 2 h showed maximal levels of hsp101 mRNA. Rice full-length hsp101 cDNA complemented yeast mutant disrupted for its own hsp104 gene by insertional mutagenesis, with efficacy that was comparable with Arabidopsis hsp101 cDNA. Electron micrographic evidence suggested that rice hsp101 cDNA in yeast is active in re-solubilizing the stress-induced protein granules in the post-stress recovery period. Rice hsp101 cDNA expression in hsp104 deficient yeast also caused recovery in tolerance against arsenite. Western analyses showed that this protein is expressed more rapidly during the stress period and retained for longer duration in the post-stress recovery period in japonica rice as compared to indica rice types. This is the first report wherein plant HSP100 protein expression is correlated to disappearance of protein granules in the yeast cells and distinct rice type-dependent protein expression patterns are reported. PMID- 12650621 TI - Ankyrin protein kinases: a novel type of plant kinase gene whose expression is induced by osmotic stress in alfalfa. AB - Interaction between Medicago spp. and Sinorhizobium meliloti leads to the development of a novel organ, the root nodule. A gene, Msapk1, encoding a novel type of plant protein kinase containing a N-terminal region with an ankyrin domain, was identified and shown to be expressed both in S. meliloti-infected and spontaneous nodules in alfalfa. This gene is not exclusively associated to nodulation since its expression was detected in other plant organs. Several genes coding for ankyrin protein kinases (APKs) were detected in various plants and animals. Three closest A. thaliana homologues of Msapk1 were identified in databases and two of them were shown to express differentially in various organs using gene-specific RT-PCR. In contrast, Southern analysis suggests that a single copy gene exists in diploid M. truncatula. By screening a M. truncatula BAC library the Mtapk1 genomic region was isolated and sequenced. Two neighbouring genes showing homologies to previously identified sequences in data banks were detected in the vicinity of the Mtapk1 gene and compared to similar regions of the three Atapk genes. The distribution of exons/introns was the same for all expressed genes of both species although Mtapk1 contained larger introns. Upon osmotic stress Msapk1 expression was induced in roots of alfalfa starting from three hours up to two days of treatment. These data suggest that Msapk1, involved in alfalfa osmotic stress responses, belongs to a novel class of plant protein kinases. PMID- 12650622 TI - Type-1 ribosome-inactivating protein from iris bulbs: a useful agronomic tool to engineer virus resistance? AB - To study the in planta antiviral activity of a type-1 ribosome-inactivating protein from iris bulbs, called IRIP, Nicotiana tabacum cv. Samsun NN was transformed with the IRIP sequence expressed under the control of the 35S cauliflower mosaic virus promoter. Molecular analysis of the transgenic plants and characterization of the purified protein revealed that the recombinant IRIP from tobacco leaves has the same molecular structure and RNA N-glycosidase activity as the native protein from iris bulbs. The tobacco transformants show no apparent phenotypic side effects indicating that ectopically expressed IRIP is not cytotoxic for tobacco cells. No induction of PR-1 could be demonstrated in the transgenic plants expressing IRIP. The in planta antiviral activity of rIRIP was assessed using a bioassay with tobacco mosaic virus. All transformed lines showed a statistically significant lower number of lesions compared to the control plants. The fortunate combination of in planta antiviral activity and lack of cytotoxicity of the ectopically expressed IRIP in transgenic tobacco renders the iris RIP an interesting and useful model for the study and exploitation of the antiviral activity of type-1 RIPs. PMID- 12650623 TI - Identification of a copper transporter family in Arabidopsis thaliana. AB - Despite copper ions being crucial in proteins participating in plant processes such as electron transport, free-radical elimination and hormone perception and signaling, very little is known about copper inward transport across plant membranes. In this work, a five-member family (COPT1-5) of putative Arabidopsis copper transporters is described. We ascertain the ability of these proteins to functionally complement and transport copper in the corresponding Saccharomyces cerevisiae high-affinity copper transport mutant. The specific expression pattern of the Arabidopsis COPT1-5 mRNA in different tissues was analyzed by RT-PCR. Although all members are ubiquitously expressed, differences in their relative abundance in roots, leaves, stem and flowers have been observed. Moreover, steady state COPT1 and COPT2 mRNA levels, the members that are most efficacious in complementing the S. cerevisiae high-affinity copper transport mutant, are down regulated under copper excess, consistent with a role for these proteins in copper transport in Arabidopsis cells. PMID- 12650624 TI - The serine acetyltransferase gene family in Arabidopsis thaliana and the regulation of its expression by cadmium. AB - Expression of the serine acetyltransferase (SAT) gene family from Arabidopsis thaliana was investigated in response to treatment with the heavy metal cadmium (Cd). A fourth member of the SAT gene family, Sat-106, was also cloned and the complete SAT gene family from A. thaliana is discussed. Northern analysis of the gene family revealed tissue-specific expression patterns for each isogene. A. thaliana plants grown under 50 microM CdCl2 for a 24 h time course were also used for northern analysis. Expression of all SAT genes was increased to some extent by Cd treatment. Sat-5 expression showed particularly high levels of induction in the leaves of treated plants and was chosen for study by in situ hybridisation. Sat-5 expression was induced in the root and stem cortex and the leaf lamella and trichomes in response to heavy metal stress. SAT and its product O-acetylserine have previously been shown to be implicated in the control of sulphate reduction and cysteine biosynthesis in plants. These results suggest that specific SAT isoforms have a role in increasing cysteine production under conditions of heavy metal stress when increased biosynthesis of glutathione and phytochelatins is required for detoxification purposes. PMID- 12650626 TI - Phototropin 1 is required for high-fluence blue-light-mediated mRNA destabilization. AB - Irradiation of etiolated wild-type Arabidopsis thaliana seedlings with a single pulse of blue (B) light with a total fluence equal to that of 1 min of sunlight causes the destabilization of nuclear-encoded Lhcb transcripts. Transcript destabilization is not observed in phototropin1 (phot1, formerly nph1) mutant seedlings, indicating that phot1 is likely the photoreceptor mediating this response. Destabilization is also absent in nph3 mutants, but occurs normally in nph4 mutants. The rates of Lhcb transcription and B low-fluence-induced Lhcb transcript accumulation are normal in phot1 seedlings, confirming that photl regulates destabilization, not a change in transcription. A similar pattern of regulation is observed for the chloroplast-encoded rbcL transcript. The lack of destabilization of a second chloroplast encoded transcript, psbD, indicates that the phot1/B-high-fluence system does not result in a general destabilization of all chloroplast transcripts. Localized sequence similarity between the Lhcb 5' UTR and the rbcL 3'-UTR suggests a similar mechanism of destabilization even though the two transcripts are located in different sub-cellular compartments. The high-fluence threshold of phot1-mediated RNA destabilization contrasts with the low-fluence threshold required for phot1-directed first-positive phototropic curvature. This study indicates that phot1, like phytochrome, can discriminate between several fluence ranges and direct responses in specific tissues or different sub-cellular compartments. PMID- 12650628 TI - Source of firearms used by students in school-associated violent deaths--United States, 1992-1999. AB - During July 1, 1992-June 30, 1999, a total of 323 school-associated violent death events occurred in the United States, resulting in 358 deaths. To guide prevention efforts, CDC examined school-associated firearm violent death events committed by students in elementary and secondary schools in the United States and determined the sources of the firearms used in these events. The findings indicate that, among the incidents for which data are available, the majority of the firearms used in these events were obtained from perpetrators' homes or from friends or relatives. The safe storage of firearms is critically important and should be continued. In addition, other strategies that might prevent firearm related injuries and deaths among students, such as safety and design changes for firearms, should be evaluated. PMID- 12650625 TI - Ozone-induced gene expression occurs via ethylene-dependent and -independent signalling. AB - Recent studies suggest that ethylene is involved in signalling ozone-induced gene expression. We show here that application of ozone increased glucuronidase (GUS) expression of chimeric reporter genes regulated by the promoters of the tobacco class I beta-1,3-glucanases (GLB and Gln2) and the grapevine resveratrol synthase (Vst1) genes in transgenic tobacco leaves. 5'-deletion analysis of the class I beta-1,3-glucanase promoter revealed that ozone-induced gene regulation is mainly mediated by the distal enhancer region containing the positively acting ethylene responsive element (ERE). In addition, application of 1-methylcyclopropene (1 MCP), an inhibitor of ethylene action, blocked ozone-induced class I beta-1,3 glucanase promoter activity. Enhancer activity and ethylene-responsiveness depended on the integrity of the GCC boxes, cis-acting elements present in the ERE of the class I beta-1,3-glucanase and the basic-type pathogenesis-related PR 1 protein (PRB-1b) gene promoters. The minimal PRB-1b promoter containing only the ERE with intact GCC boxes, was sufficient to confer 10-fold ozone inducibility to a GUS-reporter gene, while a substitution mutation in the GCC box abolished ozone responsiveness. The ERE region of the class I beta-1,3-glucanase promoter containing two intact GCC boxes confered strong ozone inducibility to a minimal cauliflower mosaic virus (CaMV) 35S RNA promoter, whereas two single-base substitution in the GCC boxes resulted in a complete loss of ozone inducibility. Taken together, these datastrongly suggest that ethylene is signalling ozone induced expression of class I beta-l,3-glucanase and PRB-1b genes. Promoter analysis of the stilbene synthase Vst1 gene unravelled different regions for ozone and ethylene-responsiveness. Application of 1-MCP blocked ethylene-induced Vst1 induction, but ozone induction was not affected. This shows that ozone induced gene expression occurs via at least two different signalling mechanisms and suggests an additional ethylene independent signalling pathway for ozone induced expression of genes involved in phytoalexin biosynthesis. PMID- 12650627 TI - Expression studies of plant genes differentially expressed in leaf and root tissues of tomato colonised by the arbuscular mycorrhizal fungus Glomus mosseae. AB - Arbuscular mycorrhizal (AM) fungi are a multifaceted group of mutualistic symbionts that are common to terrestrial ecosystems. The interaction between AM fungi and plant roots is of environmental and agronomic importance. Understanding the molecular changes within the host plant upon AM fungal colonisation is a pre requisite to a greater understanding of the mechanisms underlying the interaction. Differential mRNA display was conducted on leaf tissue of tomato plants colonised and non-colonised by the AM fungus Glomus mosseae and five putative differentially regulated cDNAs were identified. All cDNAs isolated shared high sequence similarity to known plant genes. Differential screening was initially used to establish whether the cDNAs were differentially expressed. Semi quantitative RT-PCR was used to establish gene expression patterns for all five clones within leaf and root tissue of mycorrhizal and non-mycorrhizal colonised tomato plants. Differential regulation was observed for all five cDNAs. Down regulation within the leaf tissue of mycorrhizal plants was observed for 4 out of the 5 cDNAs with an up-regulation observed only for one. Tissue specific regulation was observed for several cDNAs, with down-regulation observed in mycorrhizal leaf tissue and up-regulation observed within mycorrhizal root tissue as compared to non-mycorrhizal tissue. PMID- 12650629 TI - Progress toward poliomyelitis eradication--India, 2002. AB - Since the World Health Assembly resolved in May 1988 to eradicate poliomyelitis, the estimated global incidence of polio has decreased >99%, and three World Health Organization (WHO) regions (Americas, Western Pacific, and European) have been certified as polio-free. In 1995, India began accelerating polio eradication activities. By 2001, poliovirus circulation had been limited largely to the two northern states of Uttar Pradesh (UP) and Bihar, with 268 cases reported nationwide. However, in 2002, a major regional resurgence of polio occurred. As of January 25, 2003, a total of 1,556 cases were detected nationwide, of which 1,337 (86%) were in UP and Bihar. This report summarizes the status of polio eradication activities in India during 2002, analyzes the factors contributing to the resurgence, and describes the actions being taken to reduce poliovirus transmission. PMID- 12650630 TI - Vaccination coverage among children enrolled in Head Start programs, licensed child care facilities, and entering school--United States, 2000-01 school year. AB - The implementation of state and local requirements for vaccination before entry to Head Start programs, licensed child care facilities, and school has resulted in high vaccination levels among preschool and school children. One of the national health objectives for 2010 is to maintain > or = 95% vaccination coverage among children attending licensed child care centers and kindergarten through postsecondary school (objective 12-23). National estimates of vaccination coverage among children in Head Start programs, licensed child care facilities, and those entering school have been published each year since 1997 on the basis of reports from federally funded immunization programs (IPs) in the 50 states, five cities, eight territories, and the District of Columbia. This report summarizes data reported by states, cities, and the District of Columbia for the 2000-01 school year. Although vaccination coverage for 2000-01 appears similar to that for previous years, the number of programs reporting and the completeness of the reports are lower than in previous years and do not permit precise estimation of coverage at the national level. IPs use school data to identify undervaccinated children enrolled in Head Start programs, licensed child care facilities, and those entering school; evaluate the success of prevention programs targeting these children; and document the proportion of children whose parents claim exemptions from one or more vaccines. Plans are ongoing to assist IPs in applying successful strategies for collecting, reporting, and increasing the precision of coverage estimates for these populations. PMID- 12650631 TI - Smallpox vaccine adverse events among civilians--United States, February 25-March 3, 2003. AB - During the civilian smallpox vaccination program, CDC, the Food and Drug Administration, and state health departments are conducting surveillance for vaccine-associated adverse events. In the first stage of the program, active surveillance is being conducted for potentially life-threatening, moderate-to severe, and other serious adverse events and for vaccinia transmission to contacts of vaccinees (Table). Nonserious events are reported through passive surveillance and are expected to be underreported. This report summarizes smallpox vaccine adverse events reported among civilians vaccinated as of February 28, 2003, and among contacts of vaccinees, received by CDC from the Vaccine Adverse Event Reporting System (VAERS) as of March 3. PMID- 12650632 TI - Application of quantitative signal detection in the Dutch spontaneous reporting system for adverse drug reactions. AB - The primary aim of spontaneous reporting systems (SRSs) is the timely detection of unknown adverse drug reactions (ADRs), or signal detection. Generally this is carried out by a systematic manual review of every report sent to an SRS. Statistical analysis of the data sets of an SRS, or quantitative signal detection, can provide additional information concerning a possible relationship between a drug and an ADR. We describe the role of quantitative signal detection and the way it is applied at the Netherlands Pharmacovigilance Centre Lareb. Results of the statistical analysis are implemented in the traditional case-by case analysis. In addition, for data-mining purposes, a list of associations of ADRs and suspected drugs that are disproportionally present in the database is periodically generated. Finally, quantitative signal generation can be used to study more complex relationships, such as drug-drug interactions and syndromes. The results of quantitative signal detection should be considered as an additional source of information, complementary to the traditional analysis. Techniques for the detection of drug interactions and syndromes offer a new challenge for pharmacovigilance in the near future. PMID- 12650634 TI - Avoiding drug-induced switching in patients with bipolar depression. AB - Antidepressant-induced switching is a major risk during the treatment of bipolar depression. Despite several clinical studies, questions remain regarding both the definition of these mood switches and the most appropriate therapeutic strategy to avoid this adverse effect. This review will first briefly consider the current guidelines for the acute treatment of bipolar depression. We will then review the mechanisms of action of antidepressant and mood stabilisers, and the switches induced by various types of antidepressant treatments, or triggered by antidepressant withdrawal, as well as by atypical antipsychotics. We then will address the risk of mood switch according to the type of mood stabiliser used. The propensity to mood switches in bipolar patients is subject to individual differences. Therefore we will describe both the clinical and biological characteristics of patients prone to mood switches under antidepressant treatment. However, the clinical characteristics of the depressive syndrome may also be a key determinant for mood switches. Various data help identify the most appropriate drug management strategies for avoiding mood switches during the treatment of bipolar depression. Selective serotonin reuptake inhibitors appear to be the drugs of first-choice because of the low associated risk of mood switching. Antidepressants must be associated with a mood stabiliser and the most effective in the prevention of switches seems to be lithium. Whatever the mood stabiliser used, effective plasma levels must be ensured. The optimal duration of antidepressant treatment for bipolar depression is still an open issue - prolonged treatments after recovery may be unnecessary and may facilitate mood elation. Moreover, some mood episodes with mixed symptoms can be worsened by antidepressants pointing to the need for a better delineation of the categories of symptoms requiring antidepressant treatment. Finally, as a result of this review, we suggest some propositions to define drug-induced switches in bipolar patients, and to try to delineate which strategies should be recommended in clinical practice to reduce as far as possible the risk of mood switch during the treatment of bipolar depression. PMID- 12650635 TI - Lack of awareness of community-acquired adverse drug reactions upon hospital admission : dimensions and consequences of a dilemma. AB - OBJECTIVE: Adverse drug reactions (ADRs) are a well-known cause of hospital admission. Nevertheless a quantitative estimate of the preventability of and physicians' awareness of these reactions is lacking. STUDY DESIGN AND METHODS: Using intensive bedside and computer-assisted drug surveillance methods a 13 month prospective pharmacoepidemiological survey was carried out on patients admitted to two medical wards of the Erlangen-Nuremberg University Hospital in Erlangen, Germany. This study aimed to define the incidence of preventable and unavoidable ADRs. In addition we investigated the awareness of the physicians to ADRs at the time of admission and the rate of contraindicated pre-admission prescriptions. RESULTS: In 78 (8.5%) of 915 (10.9%) admissions a total of 102 (42 preventable) community-acquired ADRs were detected on admission. In 45 (3.8%) of the admissions ADRs led directly to hospitalisation. 56.9% of the ADRs were not recognised by the attending physician on admission. Marked correlation was found between the awareness of ADRs and their probability and severity scores (r = 0.85 and r = 0.94, respectively; p < 0.05). The most frequently detected ADRs were due to direct toxicity and secondary pharmacological effects. Idiosyncratic reactions were often missed and 18.6% of all drugs prescribed prior to admission were contraindicated. Leading the list were diuretics, analgesics/NSAIDs and antipsychotics/sedatives. CONCLUSIONS: Awareness of existing ADRs on hospital admission and appropriate prescribing prior to hospital admission require attention. Early detection of ADRs on hospital admission can be achieved by the use of computer support systems. Many ADRs could be prevented by adhering to indications and contraindications. PMID- 12650633 TI - Risks and benefits, advantages and disadvantages of levonorgestrel-releasing contraceptive implants. AB - Levonorgestrel-releasing implants are long acting contraceptives, approved for 5 years of continuous use. Two marketed systems, the six capsule Norplant use of tradenames is for product identification purposes only and does not imply endorsement. and the two rod Jadelle, have essentially equal rates of drug release, pregnancy and adverse events over 5 years of use. Randomised clinical trials and controlled cohort observations indicate that for the first 3 years, when pregnancy rates are at or almost zero, no other contraceptive system is more effective, although etonogestrel implants provide equal effectiveness. Annual pregnancy rates rise in the fifth year of continuous use but remain below 1 per 100 women. Annual pregnancy rates of Norplant users remain below 1 per 100 throughout 7 years of continuous use. Levonorgestrel implants provide low progestogen doses; 40-50 microg/day at 1 year of use, decreasing to 25-30 microg/day in the fifth year. Serum levels of levonorgestrel at 5 years are 60 65% of those levels measured at 1 month of use. Adverse effects with levonorgestrel implants are similar to those observed with progestogen only and combined oral contraceptives. Risks of ectopic pregnancy, other pregnancy complications and pelvic inflammatory disease are reduced in comparison with those of women using copper or non-medicated intrauterine devices. Risks of developing gallbladder disease and hypertension or borderline hypertension, although small, are about 1.5 and 1.8 times greater, respectively, in women using levonorgestrel implants than in women not using hormonal contraception. Other serious diseases have not been found to occur significantly more frequently in levonorgestrel implant users than in women not using hormonal contraception. The great majority of levonorgestrel implant users experience menstrual problems, but serious bleeding problems are not more frequent than in controls. Other health problems reported more frequently by levonogestrel implant users than by women not using hormonal contraception in a study of 16000 women included skin conditions, headache, upper limb neuropathies, dizziness, nervousness, malaise, minor visual disturbances, respiratory conditions, arthropathies, weight change, anxiety and non-clinical depression. Clinical depression is not more frequent in women using implants compared with those not using hormonal contraception (i.e. using intrauterine devices, sterilisation). Removal problems occur less frequently with Jadelle than with Norplant. The mean removal time for Jadelle is half that of Norplant. Levonorgestrel implants in nationally representative scientific samples, in randomised trials, and in controlled cohort studies have continuation rates as high as or higher than any other reversible contraceptive over a duration of 5 years. This would imply that the satisfaction women derive from the contraceptive effectiveness of levonorgestrel implants greatly outweighs the dissatisfaction that may accompany menstrual disturbances and other adverse effects associated with implants. PMID- 12650636 TI - Violation of homogeneity: a methodologic issue in the use of data mining tools. PMID- 12650638 TI - Periodontal disease is associated with lower antioxidant capacity in whole saliva and evidence of increased protein oxidation. AB - The aim of this cohort study was to determine whether periodontitis and gingivitis are associated with impaired salivary antioxidant status and increased oxidative injury. One hundred and twenty-nine patients attending a routine dental check-up were recruited for the study. Periodontal disease status was characterized using the Community Periodontal Index of Treatment Needs (CPITN) system. Total salivary antioxidant capacity and salivary ascorbate, urate and albumin were determined in a sample of whole unstimulated saliva. Protein carbonyl concentrations were determined as an index of oxidative injury. Patients in the lowest tertile of CPITN score exhibited decreased salivary delivery of antioxidants and specifically urate than patients in the upper tertile. Poor periodontal health was associated with increased concentrations of protein carbonyls in saliva. Women had significantly lower total antioxidant status than men, regardless of periodontal health. Periodontal disease is associated with reduced salivary antioxidant status and increased oxidative damage within the oral cavity. PMID- 12650639 TI - Localization of a highly active pool of type II phosphatidylinositol 4-kinase in a p97/valosin-containing-protein-rich fraction of the endoplasmic reticulum. AB - Different phosphoinositides are synthesized in cell membranes in order to perform a variety of functions. One of the most abundant of these lipids is phosphatidylinositol (PI) 4-phosphate (PI4P), which is formed in human eukaryotes by type II and type III phosphatidylinositol 4-kinase (PI4K II and III) activities. PI4K II activity occurs in many different subcellular membranes, although no detailed analysis of the distribution of this activity has been reported. Using density gradient ultracentrifugation, we have previously found that in A431 cells the predominant PI4K activity arises from a type II alpha enzyme that is localized to a buoyant membrane fraction of unknown origin [Waugh, Lawson, Tan and Hsuan (1998) J. Biol. Chem. 273, 17115-17121]. We show here that these buoyant membranes contain an activated form of PI4K II alpha that can be separated from the bulk of the PI4K II alpha protein in A431 and COS-7 cells. Proteomic analysis revealed that the buoyant membrane fraction contains numerous endoplasmic reticulum (ER)-marker proteins, although it was separated from the bulk of the ER, ER-Golgi intermediate compartment, transitional ER, Golgi and other major subcellular membranes. Furthermore, the majority of the cytoplasmic valosin-containing protein (VCP), an AAA+ATPase implicated in various ER, transitional ER, Golgi and nuclear functions, was almost completely localized to the same buoyant membrane fraction. Co-localization of VCP and PI4K activity was confirmed by co-immunoprecipitation. These results suggest the previously unsuspected existence of an ER-related domain in which the bulk of the cellular PI4P synthesis and VCP are localized. PMID- 12650642 TI - Emergence of modularity in genotype-phenotype mappings. AB - A novel evolutionary method that allows us to study the emergence of modularity for genotype-phenotype mapping in the course of Darwinian evolution is described. The method is based on composite epigenotypes with two parts: a binary genotype; and a mapping of genes onto phenotype characters. For such generalized epigenotypes the modularity is determined in the following intuitive way: The genes are divided into two subgroups; simultaneously with this decomposition there is defined an accompanying decomposition of the set of phenotype characters. We expect that for epigenotypes with modular structures the genes from one group will be mapped onto characters from the same group, that is, that the appearance of crosslink mappings will be maximally suppressed. A fundamental question for all of evolutionary biology (and also for evolutionary algorithms and connectionist cognitive science) is the mechanism of evolutionary emergence of modular structures. The presented explanatory model is an implementation of the assumption that variation in genotype is produced on a faster time scale than variation in the genotype-phenotype mapped part. Moreover, the evaluation of the epigenotype in the evolutionary algorithm is based on directly selectable properties (corresponding to the decomposition of the set of phenotype characters). The modularity of genotype-phenotype mapping emerges in the simulations. PMID- 12650643 TI - Learning and the evolution of language: the role of cultural variation and learning costs in the Baldwin effect. AB - The Baldwin effect has been explicitly used by Pinker and Bloom as an explanation of the origins of language and the evolution of a language acquisition device. This article presents new simulations of an artificial life model for the evolution of compositional languages. It specifically addresses the role of cultural variation and of learning costs in the Baldwin effect for the evolution of language. Results show that when a high cost is associated with language learning, agents gradually assimilate in their genome some explicit features (e.g., lexical properties) of the specific language they are exposed to. When the structure of the language is allowed to vary through cultural transmission, Baldwinian processes cause, instead, the assimilation of a predisposition to learn, rather than any structural properties associated with a specific language. The analysis of the mechanisms underlying such a predisposition in terms of categorical perception supports Deacon's hypothesis regarding the Baldwinian inheritance of general underlying cognitive capabilities that serve language acquisition. This is in opposition to the thesis that argues for assimilation of structural properties needed for the specification of a full-blown language acquisition device. PMID- 12650640 TI - MEKK3 interacts with the PA28 gamma regulatory subunit of the proteasome. AB - The proteasome is a multisubunit proteolytic enzyme comprising activator complexes bound to the 20 S catalytic core. The functions of the proteasomal activator (PA) 700 in ubiquitin/ATP-dependent protein degradation and of the PA28 alpha/beta activators in antigen presentation are well defined. However, the function of a third PA, PA28 gamma, remains elusive. We now show that mitogen activated protein kinase (MAPK)/extracellular-signal-regulated kinase (ERK) kinase kinase 3 (MEKK3), a MAPK kinase kinase (MAPKKK) involved in MAPK kinase 7 (MKK7)-c-Jun N-terminal kinase ('JNK') and MKK6-p38 signalling, can bind PA28 gamma but not PA28 alpha. In contrast, B-Raf, a MAPKKK specific for the MAPK/ERK kinase ('MEK')-ERK module, binds PA28 gamma and alpha. The PA28 gamma-binding domain of MEKK3 is located within its N-terminal regulatory domain (amino acids 1 178). Expression of MEKK3 in Cos-7 cells led to an increase in endogenous and co expressed PA28 gamma protein levels, whereas kinase-deficient MEKK3 had no effect on PA28 gamma expression. Furthermore, in vitro assays indicated that PA28 gamma was a MEKK3 substrate. MEKK3 represents the first protein kinase capable of binding and phosphorylating a PA, and provides a potential mechanism to link stress-activated protein kinase signalling with the PA28 gamma-dependent proteasome. PMID- 12650641 TI - Properties of phagocyte NADPH oxidase p47-phox mutants with unmasked SH3 (Src homology 3) domains: full reconstitution of oxidase activity in a semi recombinant cell-free system lacking arachidonic acid. AB - In an early step in the assembly of the phagocyte NADPH oxidase, p47-phox translocates from the cytosol to the membrane, mediated by engagement of the N termini of two p47-phox Src homology 3 (SH3) domains with a proline-rich region (PRR) in the p22-phox subunit of cytochrome b (558). In response to phagocyte activation, several serine residues in a C-terminal arginine/lysine-rich domain of p47-phox are phosphorylated, leading to changes in the conformation of p47 phox and exposure of its N-terminal SH3 domain that is normally masked by internal association with the arginine/lysine-rich domain. We report that triple alanine substitutions at Asp-217, Glu-218 and Glu-223 in a short sequence that links the tandem p47-phox SH3 domains unmasked the N-terminal SH3 domain, similar to the effects of aspartic acid substitutions at Ser-310 and Ser-328 in the arginine/lysine-rich region. Recombinant p47-phox proteins with mutations in either the linker region or the arginine/lysine-rich domain were active in the absence of arachidonic acid stimulation in a cell-free NADPH oxidase system consisting of recombinant p67-phox, Rac1-guanosine 5'-[gamma-thio]triphosphate and neutrophil membranes. Supplementing neutrophil membranes with phosphoinositides or other negatively charged phospholipids markedly enhanced cell-free superoxide generation by these p47-phox mutants in the absence of arachidonic acid, to levels equivalent to those generated by wild-type p47-phox following arachidonic acid activation. This enhancement may be related to recruitment to the membrane of p47-phox mediated by a novel secondary phox homology (PX) domain binding site that broadly recognizes phospholipids. No specific enhancement by specific phosphorylated phosphatidylinositols was found to suggest a dominant role for the p47-phox primary PX domain binding site. Truncated p47-phox S310D S328D lacking the C-terminal PRR was inactive in the cell-free system without arachidonic acid, but was fully active with arachidonic acid. This suggests that activation of NADPH oxidase in an arachidonate-free cell free system requires association of the p47-phox C-terminal PRR with the p67-phox C-terminal SH3 domain. PMID- 12650644 TI - Evolvable self-replicating molecules in an artificial chemistry. AB - This paper gives details of Squirm3, a new artificial environment based on a simple physics and chemistry that supports self-replicating molecules somewhat similar to DNA. The self-replicators emerge spontaneously from a random soup given the right conditions. Interactions between the replicators can result in mutated versions that can outperform their parents. We show how artificial chemistries such as this one can be implemented as a cellular automaton. We concur with Dittrich, Ziegler, and Banzhaf that artificial chemistries are a good medium in which to study early evolution. PMID- 12650645 TI - Bioinspired engineering of exploration systems for NASA and DoD. AB - A new approach called bioinspired engineering of exploration systems (BEES) and its value for solving pressing NASA and DoD needs are described. Insects (for example honeybees and dragonflies) cope remarkably well with their world, despite possessing a brain containing less than 0.01% as many neurons as the human brain. Although most insects have immobile eyes with fixed focus optics and lack stereo vision, they use a number of ingenious, computationally simple strategies for perceiving their world in three dimensions and navigating successfully within it. We are distilling selected insect-inspired strategies to obtain novel solutions for navigation, hazard avoidance, altitude hold, stable flight, terrain following, and gentle deployment of payload. Such functionality provides potential solutions for future autonomous robotic space and planetary explorers. A BEES approach to developing lightweight low-power autonomous flight systems should be useful for flight control of such biomorphic flyers for both NASA and DoD needs. Recent biological studies of mammalian retinas confirm that representations of multiple features of the visual world are systematically parsed and processed in parallel. Features are mapped to a stack of cellular strata within the retina. Each of these representations can be efficiently modeled in semiconductor cellular nonlinear network (CNN) chips. We describe recent breakthroughs in exploring the feasibility of the unique blending of insect strategies of navigation with mammalian visual search, pattern recognition, and image understanding into hybrid biomorphic flyers for future planetary and terrestrial applications. We describe a few future mission scenarios for Mars exploration, uniquely enabled by these newly developed biomorphic flyers. PMID- 12650646 TI - Artificial life and the Chinese room argument. AB - "Strong artificial life" refers to the thesis that a sufficiently sophisticated computer simulation of a life form is a life form in its own right. Can John Searle's Chinese room argument [12]-originally intended by him to show that the thesis he dubs "strong AI" is false-be deployed against strong ALife? We have often encountered the suggestion that it can be (even in print; see Harnad [8]). We do our best to transfer the argument from the domain of AI to that of ALife. We do so in order to show once and for all that the Chinese room argument proves nothing about ALife. There may indeed be powerful philosophical objections to the thesis of strong ALife, but the Chinese room argument is not among them. PMID- 12650647 TI - The United Kingdom's experience of providing health care for refugees: time for international standards? PMID- 12650648 TI - Travelers' knowledge, attitudes, and practices on prevention of infectious diseases: results from a pilot study. AB - BACKGROUND: The European Travel Health Advisory Board conducted a cross-sectional pilot survey to evaluate current travel health knowledge, attitudes, and practice (KAP) and to determine where travelers going to developing countries obtain travel health information, what information they receive, and what preventive travel health measures they employ. METHOD: Trained interviewers invited passengers at the departure gates of three international airports: London Heathrow, Paris Charles de Gaulle, and Munich to respond to a self-completion questionnaire. A total of 609 responses were collected. RESULTS: The study showed that more than one-third of travelers questioned had not sought pretravel health advice and of those who did, over 20% sought advice 14 days or less prior to travel. One-third of the respondents were aged 50 or more, and 20% had planned their trip less than 2 weeks before leaving. Only a minority were able to demonstrate that they had been immunised as per the World Health Organization or national recommendations. Respondents often misperceived both the risk of malaria at the destination and recommended preventive measures. CONCLUSIONS: The results of this pilot survey provided a valuable insight into the KAP of travelers and highlighted an important educational need among those traveling to risk destinations. Strategies are needed for raising awareness of preventable travel health issues and for raising compliance with existing recommendations. PMID- 12650650 TI - An outbreak of acute pulmonary histoplasmosis in members of a trekking trip in Martinique, French West Indies. AB - BACKGROUND: Thirteen clustered cases of American histoplasmosis, a deep mycosis caused by Histoplasma capsulatum and acquired through inhalation of airborne spores was reported. Twenty-five persons traveled in Martinique, French West Indies. Thirteen underwent trekking and passed through a mountain tunnel full of bats (tunnel group). The 12 others performed canyoning and did not go through the tunnel (control group). Fifteen days after exposure, 1 patient of the tunnel group developed fever, chills, and cough. METHODS: The index case was diagnosed in the hospital, but 12 cases where initially diagnosed as prolonged influenza. All individuals were contacted and submitted to a phone questionnaire. They were asked about eventual occurrence of influenzalike symptoms, about activities practiced, and the notion of contact with bats. All were invited to have clinical examinations, chest x-ray films, and blood samplings. Serologic testing for histoplasmosis was performed by immunodiffusion. Clinical evidence of infection with H. capsulatum was obtained in all the remaining patients of the tunnel group and in none in the control group. Symptoms occurred with an acute onset in 11 to 23 days: fever and chills, severe asthenia, headaches, digestive tract involvement, and then cough, dyspnea, hepatic involvement. Pulmonary micro- or macronodules and mediastinal adenopathies were seen on radiograph and/or computed tomography scan. RESULTS: H. capsulatum serologic tests were positive in all 13 cases with presence of specific M and or H precipitins, 5 to 13 weeks after exposure, and were negative in control group. All patients were treated with itraconazole 200 mg per day during at least 2 months. Treatment was well tolerated; patients progressively recovered. Clinical and serologic follow-up was obtained for some patients at 1 and 4 years. The present study reports the first large outbreak of histoplasmosis acquired in Martinique. CONCLUSION: Histoplasmosis still occurs and is potentially serious. In patients returning from endemic areas, presenting prolonged influenzalike symptoms, clinicians should look for previous possible exposure to Histoplasma. PMID- 12650649 TI - Incidence of health crises in tourists visiting Jamaica, west indies, 1998 to 2000. AB - BACKGROUND: Tourism is important to the Jamaican economy accounting for approximately 25% of the gross domestic product. Health problems in tourists could have significant impact on the health of the local population, the scarce health service resources, and the tourist industry. This study was conducted to identify health problems most commonly occurring in tourists visiting Jamaica and examine how these problems are managed. METHODS: Records of health problems occurring in tourists who visited principal tourist areas on the north coast from June 1998 to June 2002 were reviewed for the type of illness and how the problem was handled. The data were analyzed using Epi-Info software (Centers for Disease Control and Prevention, Atlanta, GA) and Statistical Analysis System software (SAS Institute, Cary, NC). RESULTS: Accidents were the most common health crises reported by tourists. Gastrointestinal, respiratory, and cardiovascular problems occurred less frequently. Those less than 40 years of age more frequently reported accidents or injury, gastrointestinal problems, and drug abuse, whereas respiratory and cardiovascular problems were more common among those above 40 years of age. Cardiovascular problems, drug abuse, and death were more common in men than in women. Hotel nurses handled most of the cases and were more likely to refer patients to private physicians or hospitals than to public hospitals (p <.05). Factors influencing the way the crisis was handled were age (p =.0441); who handled the crisis (p <.0001); and the method of payment (p =.0072). The factors that influenced hospitalization were gender (p =.0615); who handled the crisis at the onset (p =.0497); how the crisis was dealt with (p =.0336); and previous health problems (p =.0056). Men were more likely to be hospitalized and to be referred to a public hospital than women. Medical insurance covered the costs for 11% of tourists, and 75% paid out of pocket. CONCLUSION: The information provided by this study can be used to implement changes to reduce health problems in tourists and improve emergency health services in tourist areas. PMID- 12650651 TI - Inca Trail porters: the health of local tourism employees as a challenge for travel medicine. AB - BACKGROUND: Trekking is an activity that forms part of the increasing adventure and outdoor tourism. High altitude trekking in the Himalayas or Andes has been popular for some time. For longer treks, porters are employed to carry necessary equipment. Porters' working conditions are unfortunate and subsequent health problems considerable. Although Himalayan porters have received some attention in the press and research literature, porters on the popular Inca Trail in Peru have been neglected. In light of the growing awareness of health problems of local tourism employees, the purpose of this study was to describe Inca Trail porters' working conditions and their reports on their related health status to provide baseline information for further research and strategies for improvement. METHOD: For this descriptive study, 101 Inca Trail porters were interviewed (August/September 2001) using a structured interview schedule. RESULTS: Porters were between 17 and 68 years old; estimated body weight ranged from 50 kg to 76 kg. The usual portering job lasts for 4 days with 9 to 10 hours of carrying per day. Estimated weight of loads ranged from 20 kg to 45 kg. Major concerns were lack of fuel, clothes, shelter, and equipment but foremost the lack of sufficient food provisions. A third described their general health as poor or very poor and attributed this to work. Health complaints included respiratory infections, kidney problems, or rheumatism. Thirty-eight porters recalled injuries while on the trail and over 90% had fallen ill on the job with cold, "majurki," and stomach pain due to lack of food or cold food being named most often. Porters' demands for improvement included increased pay and appropriate and sufficient food. CONCLUSIONS: The porters' working conditions and subsequent health problems need to be addressed. A range of stakeholders is responsible for the porters' conditions and are in a position to improve current situations. Specific responsibility for health care lies with travel health professionals and local health authorities since the health of host communities has been acknowledged as an important area within travel medicine. PMID- 12650652 TI - Chemoprophylaxis use and the risk of malaria in travelers to Nepal. AB - BACKGROUND: Malaria transmission in Nepal is focal and seasonal. Based on data in returning travelers the risk of malaria is low. Sources of advice give contradictory information regarding the need for chemoprophylaxis. As a result, a degree of confusion exists among visitors. The aim of this study was to describe chemoprophylactic practices among travelers to Nepal and to document differences in advice according to its source and the country in which it was given. METHODS: A questionnaire survey of tourists attending the CIWEC Clinic Travel Medicine Center, Kathmandu between June 2000 and May 2001. Resident expatriates and indigenous Nepalese were excluded. RESULTS: Completed questionnaires were obtained from 1,303 respondents. Two hundred and eighty-eight respondents were taking chemoprophylaxis specifically for their trip to Nepal (22%), whereas 958 were not. Travelers from the United Kingdom and Denmark were significantly more likely, and those from the United States and Germany significantly less likely, to be taking chemoprophylaxis. Most travelers sought pretravel advice (71%), and all sources were more likely to advise them not to take chemoprophylaxis than to take it. However, travelers advised by a family practitioner were significantly more likely to be taking chemoprophylaxis than those advised by a travel medicine specialist. Of those taking chemoprophylaxis, 53% were doing so for a visit to the Terai alone, 33% for all areas of Nepal, and 6% for the Kathmandu Valley. Nine different chemoprophylactic regimes were in use. Six hundred and forty respondents who were not taking chemoprophylaxis had been advised that it was not necessary; 276 had made the choice themselves; and 131 had been taking chemoprophylaxis but had stopped while in Nepal. Twenty-eight of these respondents had stopped because of side effects. The most common reason for choosing not to take chemoprophylaxis was either the occurrence of side effects or the fear of them (31%). CONCLUSIONS: The variable and ultimately low risk of contracting malaria in Nepal has resulted in a lack of consensus and a wide range of opinion regarding the need for chemoprophylaxis. There is a need for clarification and tighter definition of the malaria risk faced by travelers to Nepal to avoid unnecessary chemoprophylaxis use while protecting those at significant risk. PMID- 12650653 TI - Migrants as a major risk group for imported malaria in European countries. PMID- 12650654 TI - Behavioral methods of alleviating motion sickness: effectiveness of controlled breathing and a music audiotape. AB - BACKGROUND: Behavioral countermeasures for motion sickness would be advantageous because of the side effects of antiemetic drugs, but few alternative treatments are available. The objective of this study was to compare the effectiveness of controlling breathing and listening to a music audiotape designed to reduce motion sickness symptoms, on increasing tolerance to motion-induced nausea. METHOD: Twenty-four healthy subjects were exposed to nauseogenic Coriolis stimulation on a rotating turntable under three conditions: whilst focusing on controlling breathing; listening to a music audiotape; or without intervention (control). The three conditions were performed by each subject according to a replicated factorial design at 1-week intervals at the same time of day. Ratings of motion sickness were obtained every 30 seconds. Once a level of mild nausea was reached subjects commenced controlling breathing or listened to the music audiotape. Motion was stopped after the onset of moderate nausea. RESULTS: Mean (+/- SD) motion exposure time in minutes tolerated before the onset of moderate nausea was significantly longer (p <.01) for controlling breathing (10.7 +/- 5.6 min) and longer (p <.01) for music (10.4 +/- 5.6 min) compared with control (9.2 +/- 5.9 min). CONCLUSIONS: Both controlling breathing and the music audiotape provided significant protection against motion sickness and with similar effectiveness. These nonpharmacologic countermeasures are only half as effective as standard doses of anti-motion sickness drugs, such as oral scopolamine; however, they are easy to implement and free of side effects. PMID- 12650655 TI - Tolerability of multiple vaccinations in travel medicine. AB - BACKGROUND: Due to time constraints imposed by pending departure dates of travelers, the application of multiple vaccinations is commonly practiced in pretravel counseling. However, data regarding the tolerability of schedules with simultaneous vaccinations with different products are sparse. METHOD: In order to investigate effects of this practice, a prospective study was conducted with 1,183 healthy travelers who presented prior to their departure. Standardized questionnaires covering possible side effects were collected during and after vaccination. RESULTS: Results showed an increase of the overall frequency of side effects with an increasing number of simultaneously applied vaccines. In travelers with two or more vaccinations, side effects occurred less frequently than previously published. In double vaccinations, side effects occurred in 36.7% of vaccinees, triple vaccinations in 40.3%, in more than three vaccinations in 50.0%. Subjective rating by the vaccinees showed an excellent tolerability of multiple vaccinations. CONCLUSION: Multiple vaccines can be given at the same time with limited subjective side effects. These findings may increase the acceptability of vaccinations given in combination to travelers. PMID- 12650656 TI - Pretravel health advice for asplenic individuals. PMID- 12650657 TI - Epidemiological and clinical aspects of malaria in Japan. PMID- 12650658 TI - A placebo-controlled treatment trial of Blastocystis hominis infection with metronidazole. AB - Blastocystis hominis, previously considered a harmless yeast, is now classified as a protozoan inhabiting the human intestinal tract. The pathogenicity of B. hominis remains controversial and is currently the subject of extensive debate.1- 5 As a result of the uncertainty surrounding the pathogenic role of B. hominis, large-scale treatment trials of B. hominis infection have so far been lacking. In spite of this, several drugs have been reported to be active against the parasite.6-8 The present study was carried out in order to evaluate the efficacy of metronidazole treatment in inducing clinical remission and parasitologic eradication in immunocompetent individuals with B. hominis as the only evident cause of diarrhea. PMID- 12650659 TI - Gastrointestinal Schistosomiasis japonicum infections in immigrants from the Island of Leyte, Philippines. PMID- 12650661 TI - Foreign and domestic travel and the risk of Campylobacter infection: results from a population-based sentinel surveillance scheme. PMID- 12650660 TI - Export of antimicrobial drugs by West African travelers. PMID- 12650662 TI - Upper urinary tract infection as a leading cause of fever among female travelers returning from the tropics. PMID- 12650664 TI - Selected bibliography. PMID- 12650663 TI - East African trypanosomiasis. PMID- 12650665 TI - A prevalidation study on the in vitro skin irritation function test (SIFT) for prediction of acute skin irritation in vivo: results and evaluation of ECVAM Phase III. AB - A prevalidation study sponsored by the European Centre for the Validation of Alternative Methods (ECVAM) on in vitro tests for acute skin irritation is aimed at identifying non-animal tests capable of discriminating irritants (I) from non irritants (NI), as defined according to European Union and OECD. This paper reports on Phase III for one of the methods, the skin integrity function test (SIFT), assessing the protocol performance of the SIFT, in terms of reproducibility and predictive ability, in three laboratories. The barrier function properties of excised mouse skin were determined using a set of 20 coded chemicals (10 I, 10 NI), using the endpoints of trans-epidermal water loss (TEWL) and electrical resistance (ER). The basis of the SIFT prediction model is if the ratios of the pre- and post-application values for either TEWL or ER are greater than five-fold, then the test chemical is deemed irritant (I). If the ratio of both parameters is less than five-fold then the chemical is deemed non-irritant (NI). Analysis of variance (ANOVA) indicated that the intra-lab reproducibility was acceptable but that the inter-lab reproducibility was not. Overall, the SIFT test under-predicted the irritancy of the test chemicals chosen for Phase III with an overall accuracy of only 55%. The sensitivity value (ability to correctly predict I) was only 30%. The specificity (ability to predict NI) of the test was better at 80%. A retrospective examination of the SIFT results was undertaken using Student's t-test and a significance level of P<0.05 to predict an irritant based on changes in the TEWL ratio values. This improved the predictivity of the SIFT test, giving a specificity of 60%, a sensitivity of 80% and an overall accuracy of 70%. Appropriate modifications to the prediction model have now been made and the SIFT will be re-examined in a new validation exercise to investigate the potential of this non-animal method to predict acute skin irritation potential. PMID- 12650666 TI - Arsenite-induced nitric oxide generation is cell cycle-dependent and aberrant in NBS cells. AB - Exposure to arsenic has been reported to cause DNA damage and eventually the occurrence of bladder, lung and skin cancers. A previous report has demonstrated that arsenite-induced phosphorylation of Mre11, a protein involved in the repair of DNA double strand breaks (DSBs), is M phase-dependent and requires the Nijmegen breakage syndrome (NBS) protein, NBS1 [DNA Repair 1 (2002) 137]. Furthermore, arsenite treatment arrests cells at the M phase and the cells eventually go through apoptosis [Biochemical Pharmacology 60 (2000) 771]. Here we demonstrate that arsenite treatment enhances the generation of nitric oxide (NO), and that the enhanced NO generation is dominant at the G2/M phase. Arsenite induced NO generation is impaired in DSB repair-defective NBS cells, but not in NBS1-reconstituted NBS cells, suggesting NBS1 is required for effective NO generation. In summary, our study showed, for the first time, that arsenite induced NO generation is cell-cycle- and NBS1-dependent. PMID- 12650667 TI - Growth inhibition of intestinal bacteria and mutagenicity of 2-, 3-, 4 aminobiphenyls, benzidine, and biphenyl. AB - 2-Aminobiphenyl (2-ABP), 3-aminobiphenyl (3-ABP) and 4-aminobiphenyl (4-ABP), but not benzidine (Bz) and biphenyl (Bp), were found to be inhibitory to the growth of human intestinal bacteria Bifidobacterium infantis ATCC 15697, B. bifidium ATCC 11863, Clostridium perfringens ATCC 13124, Escherichia coli ATCC 25922, E. coli ATCC 35218, Enterobacter cloacae ATCC 13047 and Salmonella typhimurium TA98, TA100, YG1041 at 10-200 microg/ml in culture broth. Bacteroides distasonis ATCC 8503, B. fragilis ATCC 25285, B. theataiotaomicron ATCC 29741, C. paraputrificum ATCC 26780, C. clostridiiforme ATCC 25537, Lactobacillus acidophilus ATCC 4356 and Enterococcus faecium ATCC 19434 were not inhibited by the above mentioned compounds in concentrations up to 200 microg/ml. The Ames Salmonella/microsome assay was employed to test the mutagenicity of the above-mentioned compounds using strains TA98 and TA100 in the presence and absence of Aroclor 1254-induced rat S9 mix. It was found that 4-ABP was mutagenic to both TA98 and TA100, and Bz was mutagenic to TA98 in the presence of rat S9 mix. 2-Aminobiphenyl, 3-ABP, and Bp were not mutagenic to both strains tested. 2-Aminobiphenyl and 3-ABP are chemical isomers of 4-ABP and are as strong as 4-ABP in inhibiting the growth of intestinal bacteria but not as mutagenic as 4-ABP. Evidence suggested that the mechanism of growth inhibition is not involved with the interaction of DNA that causes mutations, but rather on the electron transport system of these organisms. PMID- 12650668 TI - Role of reactive oxygen species in organophosphate insecticide phosalone toxicity in erythrocytes in vitro. AB - Reactive oxygen species (ROS) caused by organophosphates may be involved in the toxicity of various pesticides. Therefore, in this study we aimed to investigate how an organophosphate insecticide, phosalone, affects lipid peroxidation (LPO) and the antioxidant defence system in vitro. For this purpose, the effects of various doses of phosalone on LPO and the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) in erythrocytes were studied. Each phosalone dose was incubated with a previously prepared erythrocyte sample at +4 degrees C for 0, 60 and 180 min. After incubation, the levels of malondialdehyde (MDA) and the activities of SOD, GSH-Px and CAT were determined. Phosalone caused an increase in MDA formation and a decrease in the activities of SOD, GSH-Px and CAT. However, these effects were seen only at extremely high concentrations of phosalone and these concentrations were in the lethal range. Therefore, we suggest that ROS may not involve in the toxic effects of the pesticidal use of phosalone in low concentrations. PMID- 12650669 TI - Methyl 2-(2-chloroethylaminocarbonyl)diazenecarboxylate SB-166 inhibits the growth of different tumour cell lines, including drug-resistant sublines. AB - Recently we synthesized new drugs, diazenecarboxamides (shortly diazenes), that were cytotoxic for several tumour cell lines. Because the solubility and biological activity of these drugs was relatively low, new compounds have been synthesized. In the present study we examined the cytotoxic effect of nine compounds: an imidazolidin-2-one (SB-282: methyl 5-benzoyl-3-(2-chloroethyl)-2 oxo-4-phenyl-2,3-dihydro-1H-imidazol-1-ylcarbamate), two diazenecarboxamides (UP 140: N-phenyl-2-(2-quinolinyl)diazenecarboxamide; JK-1090: N-(4-iodophenyl)-2-(2 pyridinyl)diazenecarboxamide), two aminocarbonyl substituted diazenecarboxylates (SB-178: methyl 2-[(cyclohexylamino)carbonyl]diazenecarboxylate; SB-166: methyl 2 [[(2-chloroethyl)amino]carbonyl]diazenecarboxylate) and four diazenedicarboxamides (SB-410: N(1)-(2-chloroethyl)-N(2)-(2-pyridinylmethyl)-1,2 diazenedicarboxamide; SB-472: N(1)-(2-chloroethyl)-N(2)-(4-isopropylphenyl)-1,2 diazenedicarboxamide; SB-503: N(1)-(4-sec-butylphenyl)-N(2)-(2-chloroethyl)-1,2 diazenedicarboxamide; SB-474: N(1)-(4-tert-butylphenyl)-N(2)-(2-chloroethyl)-1,2 diazenedicarboxamide). Using a modified colorimetric MTT assay, their cytotoxicity was determined on eight human cell lines: laryngeal carcinoma parental and two drug-resistant cell lines, glioblastoma parental and drug resistant cell lines, cervical carcinoma parental and drug-resistant cell lines and breast adenocarcinoma cells. Results show that diazene SB-166 was very effective, reducing significantly the cell survival of all eight examined cell lines, including four drug-resistant cell lines. Compound SB-410 was cytotoxic for all examined cell lines, but mostly only in the highest concentration. Other compounds were not significantly cytotoxic to any of the treated cell lines. Our results, especially those obtained on drug-resistant cells, encourage further research on compound SB-166 as a potential anticancer drug. PMID- 12650670 TI - Lack of inhibition of endothelial nitric oxide synthase in the isolated rat aorta by doxorubicin. AB - Besides inducing cardiotoxicity, doxorubicin also affects the vasculature. Recent observations in cultured endothelial cells indicated that the endothelial form of nitric oxide synthase might be inhibited by doxorubicin thereby seriously interfering with vascular function. We have investigated the effect of doxorubicin on the relaxation induced by the muscarinic agonist carbachol in the isolated rat aorta. It was found that doxorubicin at concentrations up to 50 microM does not alter the relaxant response to carbachol. Direct measurement of nitrite, the metabolite of NO*, by the Griess assay confirmed our observation that NO*)production is not inhibited by doxorubicin. PMID- 12650671 TI - Apoptotic effect in the glioma cells induced by specific protein extracted from Okinawa Habu (Trimeresurus flavoviridis) venom in relation to oxidative stress. AB - Okinawa Habu (Trimeresurus flavoviridis) venom is well known for its toxic efficacy, from which one kind of specific protein, Okinawa Habu apoxin protein-1 (OHAP-1) has been extracted. The purpose of this study was to investigate whether OHAP-1 could induce apoptosis in some glioma cells, and if so, to elucidate the possible mechanism involved. Three malignant glioma cell lines were tested. The malignant glioma cell lines were rat C6 and human RBR 17T, U251. OHAP-1 inhibited growth of all cell lines. Whether or not the apoptosis had been induced was determined by using DNA gel electrophoresis, DNA flow cytometry and TUNEL assay. After OHAP-1 treatment, DNA fragmentation, an increase in the percentage of subdiploid DNA content, and TUNEL positive cells were found in the C6, RBR17T, and U251 cells. Furthermore, OHAP-1 showed L-amino acid oxidase (LAAO) activity. In order to study the mechanism of apoptosis induced by OHAP-1, the changes of intracellular reactive oxygen species (ROS) were measured using flow cytometry, and the expression of p53 protein was examined using immunohistochemistry. OHAP-1 was found to generate ROS and increase the expression of p53 protein in glioma cells. The inhibiting effect of OHAP-1 on three tested cells was reversed when an antioxidant of either catalase or reduced glutathione (GSH) was added; its apoptotic effect correspondingly became weaker. In this study, the apoptotic effect of OHAP-1 on some malignant glioma cells was confirmed, and it could be that this effect might be mediated through promoting the generation of intracellular ROS and p53 protein expression in glioma cells. It was suggested that OHAP-1 is promising as a potential candidate for clinical tumor therapy. PMID- 12650672 TI - Homology modelling of CYP2A6 based on the CYP2C5 crystallographic template: enzyme-substrate interactions and QSARs for binding affinity and inhibition. AB - The results of homology modelling of the human P450 enzyme CYP2A6, based on the CYP2C5 crystallographic template structure are reported. A substantial number of selective substrates of the CYP2A6 enzyme fit the putative active site in a manner that is consistent with their known metabolites. Moreover, the evidence from site-directed mutagenesis experiments is in accordance with the current model, particularly in relation to complementary amino acid contacts within the haem environment. The binding of substrates is rationalized in terms of QSAR analyses and from a consideration of the contributory factors affecting the binding affinity. The latter approach appears to represent a highly correlated (R=0.99) method for estimating the relative strength of enzyme-substrate binding within CYP2A6-selective compounds, albeit within a fairly limited dataset of substrates. PMID- 12650673 TI - Characterisation of the connexin32 promoter and changes in response element complexes in rat liver and hepatocytes during culture associated with oxidative stress. AB - Hepatic gap junctional intercellular communication (GJIC), mediated principally by connexin 32, provides a mechanism for regulating multicellular activities between neighbouring cells. The control of Cx32 gene expression at the transcriptional level has been investigated in rat liver tissue and in primary rat hepatocytes during culture. Several response elements have been identified and characterised using the electrophoretic mobility shift assay. Nuclear protein extract prepared from rat primary hepatocytes cultured for 2 h gave a larger number of DNA-protein complexes than observed with extracts from liver in vivo, including complexes containing Sp1. In contrast, nuclear extracts prepared from primary rat hepatocytes cultured for 96 h, and subject to oxidative stress, gave altered DNA-protein complexes when compared to those from hepatocytes cultured for 2 h. These results indicate that culture conditions, known to cause a loss of connexin expression, can modulate the transcription of Cx32 in hepatocytes by affecting the regulatory trans/cis-interactions of redox-sensitive zinc finger proteins within the promoter. PMID- 12650674 TI - Precision-cut tissue slices from transgenic mice as an in vitro toxicology system. AB - In these experiments precision-cut tissue slices from two existing transgenic mouse strains, with transgenes that couple promoting or binding elements to a reporter protein, were used for determination of reporter induction. This approach combines the power of transgenic animals with the practicality of in vitro systems to investigate the biological impact of xenobiotics. Additionally, the normal cellular architecture and heterogeneity is retained in precision-cut tissue slices. Two transgenic mouse strains, one of which couples the promoting region of CYP 1A1 to beta-galactosidase, and another which couples two forward and two backward 12-O-tetradecanoyl phorbol-13-acetate (TPA) repeat elements (TRE) to luciferase (termed AP-1/luciferase), were used to determine the feasibility of this approach. Precision-cut kidney and liver slices from both transgenic strains remain viable as determined by slice K(+) ion content and LDH enzyme release. Liver slices harvested from the CYP 1A1/beta-galactosidase transgenic mice exhibit a 14-fold increase in beta-galactosidase activity when incubated with beta-napthoflavone for 24 h. Kidney and liver slices obtained from the AP-1/luciferase transgenic mice demonstrate induction of luciferase (up to 2.5-fold) when incubated with phorbol myristate acetate (PMA or TPA) up to 4 h. These data indicate that precision-cut tissue slices from transgenic mice offer a novel in vitro method for toxicity evaluation while maintaining normal cell heterogeneity. PMID- 12650675 TI - Optimisation of cell-based assays for medium throughput screening of oxidative stress. AB - Identification and development of cell-based assays adapted to medium throughput screening requirements is important when screening chemicals for their potential toxic properties. We describe here rapid fluorometer-based and spectrophotometer based microplate assays which allow for the evaluation of oxidative stress in hepatocyte cell cultures by measurement of three markers: production of hydroperoxide assessed by the DCFH-DA probe, cellular antioxidant status by measurement of reduced glutathione and glutathione peroxidase activity and cytotoxicity and mitochondrial damage by evaluation of the mitochondrial transmembrane potential with rhodamine 123 fluorescent dye. The assays described here are rapid, simple and inexpensive, which are desirable when setting up screening assays. This system should be useful in selecting candidate compounds during the pre-development phase of agrochemicals or pharmaceuticals. It should also be of interest for retrospective and explanatory studies of mechanisms underlying toxicity observed in vivo. PMID- 12650676 TI - Evaluation of the skin sensitization potential of chemicals using expression of co-stimulatory molecules, CD54 and CD86, on the naive THP-1 cell line. AB - It has been known that dendritic cells (DCs) including Langerhans cells (LCs) play a critical role in the skin sensitization process. Many attempts have been made to develop in vitro sensitization tests that employ DCs derived from peripheral blood mononuclear cells (PBMC-DC) or CD34+ hematopoietic progenitor cells (CD34+ HPC) purified from cord blood or bone marrow. However, the use of the DCs in in vitro methods has been difficult due to the nature of these cells such as low levels in the source and/or donor-to-donor variability. In our studies, we employed the human monocytic leukemia cell line, THP-1, in order to avoid some of these difficulties. At the start, we examined whether treatment of the cells with various cytokines could produce DCs from THP-1. Treatment of THP-1 cells with cytokines such as GM-CSF, IL-4, TNF-alpha, and/or PMA did induce some phenotypic changes in THP-1 cells that were characteristic of DCs. Subsequently, responses to a known sensitizer, dinitrochlorobenzene (DNCB), and a non sensitizer, dimethyl sulfoxide (DMSO) or sodium lauryl sulfate (SLS), on the expression of co-stimulatory molecules, CD54 and CD86, were examined between the naive cells and the cytokine-treated cells. Interestingly, the naive THP-1 cells responded only to DNCB and the response to the sensitizer was more distinct than cytokine-treated THP-1 cells. Similar phenomena were also observed in the human myeloid leukemia cell line, KG-1. Furthermore, with treatment of DNCB, naive THP 1 cells showed augmented expression of HLA, CD80 and secretion of IL-1 beta. The response of THP-1 cells to a sensitizer was similar to that of LCs/DCs. Upon demonstrating the differentiation of monocyte cells in our system, we then evaluated a series of chemicals, including known sensitizers and non-sensitizers, for their potential to augment CD54 and CD86 expression on naive THP-1 cells. Indeed, known sensitizers such as PPD and 2-MBT significantly augmented CD54 and CD86 expression in a dose-dependent manner while non-sensitizers, such as SLS and methyl salicylate (MS), did not. To note, the metal allergens such as (NH(4))(2)[PtCl(4)], NiSO(4) and CoSO(4) augmented significantly only CD54 expression. Taking advantage of a cultured cell line, measurement of the co stimulatory molecules, CD54 and CD86, on naive THP-1 cells following chemical exposure shows promise for the development of a simple, short-term in vitro sensitization test. PMID- 12650677 TI - Intraspecific relationship analysis by DNA markers and in vitro cytotoxic and antioxidant activity in Eleutherococcus senticosus. AB - To analyse genetic relationships and intraspecific variation within Eleutherococcus senticosus, the polymerase chain reaction (PCR) was performed on total genomic DNAs of 10 Eleutherococcus collections. Ten primers were used for amplification, yielding 106 bands, of which 87 were polymorphic. The genetic diversity and genetic distance among 10 collections of Eleutherococcus species were used to describe the dendrogram showing the phylogenic relationship. The 10 collections were classified into two groups (groups I and II) at a similarity coefficient of 0.50. Group I included E. senticosus from Bukhaedo (Japan), E. sessliliflorus from Youngwal (Korea), E. seoulense and E. chiisanesis, while group II included several internal and Russian collections. The range of polymorphism was from 66.7 to 90.9% in the 87 amplified polymorphic DNA fragments. The similarity value of all collections ranged from 0.41 to 0.92, and the average genetic distance was 0.61. Thus, RAPD analysis was useful in determining genetic relatedness among collections and in identifying different genotypes of E. senticosus and other Eleutherococcus species. Also, the biological activity on DPPH radical scavenging, antilipid peroxidation in rat liver microsomes and cytotoxic sulforhodamine B (SRB) assay was evaluated using root extracts of E. senticosus, Odaesan, Korea. Ethyl acetate and n-butanol fractionation revealed strong antioxidant against scavenging on DPPH free radical and also ethyl acetate fractionation exhibited high antilipid peroxidative activities. In the cytotoxic effects were evaluated on seven human cancer cell lines, the values of 50% growth inhibition (GI(50)) were mostly below 30 microg/ml for crude extracts to be considered as significantly active. PMID- 12650678 TI - Novel yeast bioassay system for detection of androgenic and antiandrogenic compounds. AB - Recently, certain environmental endocrine disrupters have shown to act as antiandrogens. This suggests that environmental antiandrogens may also be crucial contributors to the increasing incidence of male reproductive abnormalities, requesting the screening and classification of antiandrogenic chemicals. Here, we report the development of a rapid, simple and effective yeast detection system for androgenic and antiandrogenic compounds, which is based on the yeast two hybrid protein interaction. A yeast strain, ARhLBD-ASC1, was established by co transformation of yeast cells harboring a lacZ reporter plasmid with two vectors expressing each of LexA fused hinge-ligand binding domain (hLBD) of androgen receptor (AR) and B42 fused ASC-1 that interacts with the AR-hLBD in an androgen dependent manner. In this yeast strain, androgens, but not other hormones, strongly stimulated the beta-galactosidase activity in a dose-dependent manner. The AR antagonists flutamide, cyproterone acetate and spironolactone, and environmental antiandrogens p,p'-DDE and vinclozolin all inhibited the response of the yeast cells to 10 nM testosterone, qualitatively similar to their inhibition reported in mammalian cell systems. Furthermore, the bioassay can be performed with the simple X-gal staining on microtiter plates, suggesting this system as a powerful tool for practical and efficient screening of environmental compounds for their androgenic and antiandrogenic activities. PMID- 12650679 TI - Hormonal aspects of schizophrenias: an overview. AB - Although the lifetime risk and prevalence of schizophrenia are comparable for men and women, gender differences occur in various aspects of the disease, including age of onset, pathophysiology, symptoms, course, and response to treatment. These gender differences strongly suggest a key role played by gonadal hormones and their interactions with neurotransmitters. They may also suggest a key role for future applications of specific estrogens for improved treatment of schizophrenics.'Atypical' antipsychotics definitely improved the benefit/risk ratio of treatment of schizophrenic patients. However, they shift the reasons for noncompliance from extrapyramidal symptoms to hormonally related adverse effects, mostly weight gain and impaired sexual functions (which occur in men and women, but cause noncompliance mostly in men). Diabetes, dyslipidemia, and decreased bone mineral density, as well as some other adverse effects are more 'silent' but their long-term effects are detrimental. 'Hormone-friendlier' interventions might be needed. PMID- 12650680 TI - Gender differences in schizophrenia. AB - Sex differences in schizophrenia can be caused by the disease process itself, by genetic and hormonal differences, by differences in the maturation and morphology of the brain and in age- and gender-specific behavioural patterns. These hypotheses will be tested on the major results reported in the literature as well as on different levels (epidemiology, risk factors, animal experiments, a controlled clinical study) on data from the ABC Schizophrenia Study. Symptomatology, lifetime risk and symptom-related course of illness-the latter without consideration of age-show no gender differences. However, until menopause illness onset is delayed and severity of illness is reduced by oestrogen on the level of gene expression and transmitter functioning. Oestrogen has an antagonistic effect on the-familial or exogenous-predisposition to illness. As a result, the age distribution of onset and the severity of first-episode illness in young men and post-menopausal women differ from the normal. First intervention trials with oestrogen substitution of neuroleptic therapy have demonstrated antipsychotic effects. The poorer social course of schizophrenia in men than in premenopausal women is accounted for by men's lower level of social development at illness onset and the subsequent impediment of their further development. Men's socially adverse illness behaviour, too, is a contributing factor. Scarcity of the knowledge of differences in the development, morphology and functioning of the male and female brain does not yet allow any definitive conclusions about gender differences in schizophrenia. PMID- 12650681 TI - Prevalence of hyperprolactinemia in schizophrenic patients treated with conventional antipsychotic medications or risperidone. AB - OBJECTIVE: The prevalence of hyperprolactinemia during treatment with conventional antipsychotic drugs or risperidone is under-recognized and requires further investigation. This open-label study was designed to determine the extent of this potential problem in a routine clinical setting. METHODS: Four hundred and two adult inpatients or outpatients with a diagnosis of schizophrenia, schizophreniform disorder, or schizoaffective disorder were studied in a 1-day, point prevalence trial. Neither clinicians nor patients had any prior knowledge of serum prolactin levels or any potential associated adverse events, and patients were required to have been treated with a conventional antipsychotic drug or risperidone for a minimum of 3 months prior to study entry. Patients taking concomitant medications known to elevate prolactin were excluded. Rigorous assessment of serum prolactin was performed to estimate the prevalence rate of hyperprolactinemia, defined as a level above the upper limit of normal (>18.77 ng/ml for males, and >24.20 ng/ml for females). Patients were stratified within antipsychotic treatment by gender and, for females, by menopausal status. RESULTS: Serum prolactin was obtained from 147 females (age range: 21-69 years; mean age=44.51 years) and 255 males (age range: 18-66 years; mean age=40.76 years). The prevalence of hyperprolactinemia among women of reproductive age (n=90) was 65.6% (mean serum prolactin=69.0 ng/ml), and among postmenopausal women (n=51), it was 45.1% (mean serum PRL=49.0 ng/ml). The prevalence of hyperprolactinemia across all males (n=255) was 42.4% (mean serum PRL= 32.4 ng/ml). The prevalence of hyperprolactinemia among females taking risperidone (N=42) was 88% versus 47.6% of those taking conventional antipsychotic drugs (N=105), with 48% of those females of reproductive age on risperidone experiencing abnormal menstrual cycles (secondary amenorrhea, oligomenorrhea, or polymenorrhea). Of all premenopausal females with hyperprolactinemia, 31.6% had estradiol levels 15 years) of combined therapy, although this falls following HRT cessation suggesting a growth-promoting effect. Randomised evidence demonstrates that the efficacy of anti-estrogens, aromatase inhibitors and raloxifene in the treatment and chemoprevention of breast cancer are restricted to women with oestrogen receptor positive (ER +ve) disease; however, HRT has not been associated conclusively with a predominance of hormone sensitive breast cancer. Despite stimulating the breast cancer cell growth, HRT has not been shown to increase breast cancer recurrence or mortality when prescribed to breast cancer survivors experiencing oestrogen deficiency symptoms and randomised trials have been recommended and commenced. In conjunction with controlled breast cancer trials demonstrating a therapeutic benefit of high dose estrogens and interest in the use of additive oestrogen therapy in patients developing resistance to oestrogen deprivation, the dogma that HRT is an absolute contra-indication following diagnosis is challenged. PMID- 12650708 TI - 17 beta-hydroxysteroid dehydrogenases and cancers. AB - 17 beta-Hydroxysteroid dehydrogenases (17HSDs) catalyze the interconversions between active 17 beta-hydroxysteroids and less-active 17-ketosteroids thereby affecting the availability of biologically active estrogens and androgens in a variety of tissues. The enzymes have different enzymatic properties and characteristic cell-specific expression patterns, suggesting differential physiological functions for the enzymes. Epidemiological and endocrine evidence indicate that estrogens play a key role in the etiology of breast cancer while androgens are involved in mechanisms controlling the growth of prostatic cells, both normal and malignant. Recently, we have developed, using LNCaP prostate cancer cell lines, a cell model to study the progression of prostate cancer. In the model LNCaP cells are transformed in culture condition to more aggressive cells, able to grow in suspension cultures. Our results suggest that substantial changes in androgen and estrogen metabolism occur in the cells during the process. These changes lead to increased production of active estrogens during transformation of the cells. Data from studies of breast cell lines and tissues suggest that the oxidative 17HSD type 2 may predominate in human non-malignant breast epithelial cells, while the reductive 17HSD type 1 activity prevails in malignant cells. Deprivation of an estrogen response by using specific 17HSD type 1 inhibitors is a tempting approach to treat estrogen-dependent breast cancer. Our recent studies demonstrate that in addition to sex hormone target tissues, estrogens may be important in the development of cancer in some other tissues previously not considered as estrogen target tissues such as colon. Our data show that the abundant expression of 17HSD type 2 present in normal colonic mucosa is significantly decreased during colon cancer development. PMID- 12650710 TI - Phytoestrogens for hormone replacement therapy? AB - Due to some severe side effects "classical" hormone replacement therapy (HRT) is currently being challenged by a therapy with phytoestrogens. Particularly soy and red clover derived isoflavones are advertised as selective estrogen receptor modulators (SERMs) with only desired and no undesired estrogenic effects. Evidence that this is the case however is scarce. Most studies investigating climacteric complaints did not find beneficial effects. A proposed beneficial effect on mammary cancer is unproven. The majority of studies however indicate an antiosteoporotic effect of isoflavones, while putative beneficial effects in the cardiovascular system are questionable due to the fact that estradiol which--like isoflavones--increase HDL and decrease LDL concentrations appear not to prevent arteriosclerosis in the human. In the urogenital tract, including the vagina, soy and red clover derived isoflavones are without effects. Cimicifuga racemosa extracts are traditionally used for the treatment of climacteric complaints. Evidence is now available that the yet unknown compounds in Cimicifuga racemosa extracts prevent climacteric complaints and may also have antiosteoporotic effects. PMID- 12650711 TI - Endometriosis: the pathophysiology as an estrogen-dependent disease. AB - Endometriosis, defined as the presence of endometrial glands and stroma outside of the uterine cavity, develops mostly in women of reproductive age and regresses after menopause or ovariectomy, suggesting that the growth is estrogen-dependent. Indeed, the lesions contain estrogen receptors (ER) as well as aromatase, an enzyme that catalyses the conversion of androgens to estrogens, suggesting that local estrogen production may stimulate the growth of lesions. The expression patterns of ER and progesterone receptors in endometriotic lesions are different from those in the eutopic endometrium. Moreover, estrogen metabolism, including the expression pattern of aromatase and the regulation of 17 beta-hydroxysteroid dehydrogenase type 2 (an enzyme responsible for the inactivation of estradiol to estrone), is altered in the eutopic endometrium of women with endometriosis, adenomyosis, and/or leiomyomas compared to that in the eutopic endometrium of women without disease. Immunostaining for P450arom in endometrial biopsy specimens diagnosed these diseases with sensitivity and specificity of 91 and 100%, respectively. This is applicable to the clinical diagnosis of endometriosis. The polymorphisms in the ER-alpha gene, the CYP19 gene encoding aromatase, and several other genes are associated with the risk of endometriosis. Studies of these will lead to better understandings of the etiology and pathophysiology of endometriosis. PMID- 12650712 TI - Pros and cons of existing treatment modalities in osteoporosis: a comparison between tibolone, SERMs and estrogen (+/-progestogen) treatments. AB - Tibolone, selective estrogen receptor modulators (SERMs) like tamoxifen and raloxifene, and estrogen (+/-progestogen) treatments prevent bone loss in postmenopausal women. They exert their effects on bone via the estrogen receptor (ER) and the increase in bone mass is due to resorption inhibition. The effect of SERMs on bone mineral density is less than that with the other treatments, but the SERM raloxifene still has a positive effect on vertebral fractures. In contrast to tibolone and estrogens (+/-progestogen), SERMs do not treat climacteric complaints, whilst estrogen plus progestogen treatments cause a high incidence of bleeding. Estrogen plus progestogen combinations have compromising effects on the breast. Tibolone and SERMs do not stimulate the breast or endometrium. Unlike SERMs, tibolone does not possess antagonistic biological effects via the ER in these tissues. Estrogenic stimulation in these tissues is prevented by local metabolism and inhibition of steroid metabolizing enzymes by tibolone and its metabolites. SERMs and estrogen (+/-progestogen) treatments increase the risk of venous thromboembolism (VTE), whilst estrogen (+/ progestogen) combinations have unwanted effects on cardiovascular events. So far, no detrimental effects of tibolone have been observed with respect to VTE or cardiovascular events. The clinical profile of tibolone therefore has advantages over those of other treatment modalities. It is also clear that tibolone is a unique compound with a specific mode of action and that it belongs to a separate class of compounds that can best be described as selective, tissue estrogenic activity regulators (STEARs). PMID- 12650713 TI - Non-genomic aldosterone action: from the cell membrane to human physiology. AB - According to the traditional model, steroid hormones bind to intracellular receptors and subsequently modulate transcription and protein synthesis, thus triggering genomic events finally responsible for delayed effects. In addition, very rapid effects of steroids mainly affecting intracellular signaling have been widely recognized which are clearly incompatible with the genomic model. These rapid, non-genomic steroid actions are likely to be transmitted via specific membrane receptors. Evidences for non-genomic steroid effects and distinct receptors involved are now presented for all steroid groups including vitamin D(3) and thyroid hormones. Mechanisms of action are being studied with regard to signal perception and transduction involved, and for various steroids including aldosterone a patchy sketch of a membrane receptor/second messenger cascade shows up being not essentially dissimilar to cascades involved in catecholamine or peptide hormone action. Aside non-classical membrane receptors with a high affinity for aldosterone, these effects involve phospholipase C, phosphoinositide turnover, intracellular pH and calcium, protein kinase C and tyrosine kinases. Increasing evidence is being accumulated for rapid physiological responses in humans, e.g. at the level of circulatory or metabolic effects, rendering clinical significance to these rapid actions. PMID- 12650714 TI - Progesterone receptor interacting coregulatory proteins and cross talk with cell signaling pathways. AB - Progesterone receptor (PR) is a member of the nuclear receptor family of ligand dependent transcription activators and is expressed as two different sized proteins from a single gene; PR-A and PR-B. The two PR isoforms are identical in their DNA binding domains (DBD) and C-terminal ligand binding domains (LBD), differing only in the N-terminal domain that is truncated in PR-A. PR also contains two autonomous transcription activation domains (AD), ligand-dependent AF-2 in the C-terminus and constitutive AF-1 in the N-terminus. AF-2 is highly conserved and a family of p160 coactivators that interacts with and mediates the activity of AF-2 has been well characterized. By contrast the N-terminal domain and AF-1 are not conserved and little is known about AF-1 coactivators. The N terminal domain is functionally important as it is required for full transcription activity of PR and is responsible for the distinct activities of the two PR isoforms, as well as cell and promoter specific functions of PR. This paper describes our efforts to identify PR N-terminal interacting coregulatory proteins. We summarize our work on the role of jun dimerization protein-2 (JDP-2) as an AF-1 coactivator of PR. JDP-2, initially defined as a repressor of jun and other bZIP transcription factors, also functions as a potent PR selective coactivator. JDP-2 lacks an intrinsic activation domain and through association with the DBD, we propose that JDP-2 potentiates AF-1 by recruiting other coactivators independent of AF-2 and p160 pathways. We also discovered that PR contains an SH3 domain interaction motif in the N-terminus that mediates interaction with Src tyrosine kinases and other signaling molecules. This interaction mediates rapid progesterone activation of Src/MAP K signaling pathways and defines a molecular mechanism for some of the rapid non-genomic actions of progesterone. PMID- 12650715 TI - Rapid effects of estrogen on G protein-coupled receptor activation of potassium channels in the central nervous system (CNS). AB - Estrogen rapidly alters the excitability of hypothalamic neurons that are involved in regulating numerous homeostatic functions including reproduction, stress responses, feeding and motivated behaviors. Some of the neurons include neurosecretory neurons such as gonadotropin-releasing hormone (GnRH) and dopamine neurons, and local circuitry neurons such as proopiomelanocortin (POMC) and gamma aminobutyric acid (GABA) neurons. We have elucidated several non-genomic pathways through which the steroid alters synaptic responses in these hypothalamic neurons. We have examined the modulation by estrogen of the coupling of various receptor systems to inwardly-rectifying and small-conductance, Ca(2+)-activated K(+) (SK) channels using intracellular sharp-electrode and whole-cell recording techniques in hypothalamic slices from ovariectomized female guinea pigs. Estrogen rapidly uncouples mu-opioid receptors from G protein-gated inwardly rectifying K(+) (GIRK) channels in POMC neurons and GABA(B) receptors from GIRK channels in dopamine neurons as manifested by a reduction in the potency of mu opioid and GABA(B) receptor agonists to hyperpolarize their respective cells. This effect is blocked by inhibitors of protein kinase A (PKA) and protein kinase C (PKC). In addition, after 24h following steroid administration in vivo, the GABA(B)/GIRK channel uncoupling observed in GABAergic neurons of the preoptic area is associated with reduced agonist efficacy. Conversely, estrogen enhances the efficacy of alpha(1)-adrenergic receptor agonists to inhibit apamin-sensitive SK currents in these preoptic GABAergic neurons, and does so in both a rapid and sustained fashion. Finally, we observed a direct, steroid-induced hyperpolarization of GnRH neurons. These findings indicate a richly complex yet coordinated steroid modulation of K(+) channel activity in hypothalamic (POMC, dopamine, GABA, GnRH) neurons that are involved in regulating numerous homeostatic functions. PMID- 12650716 TI - Estrogen can protect neurons: modes of action. AB - The female sex hormone estrogen is a steroid that activates estrogen receptors which may act as nuclear transcription factors. In addition to its well-known functions in reproductive tissue, estrogen and its receptors may also act in various regions throughout the body including the brain. Recently, the neuroprotective functions of estrogen have been investigated in greater detail. Besides acting as a powerful neuroprotective antioxidant molecule estrogen may activate the transcription of neuroprotective genes directly and may cross-talk with various intracellular signaling processes. The various modes of action of estrogen via receptor activation and in a non-receptor fashion are currently under intensive investigation. PMID- 12650717 TI - Basis of progesterone protection in spinal cord neurodegeneration. AB - Progesterone neuroprotection has been reported in experimental brain, peripheral nerve and spinal cord injury. To investigate for a similar role in neurodegeneration, we studied progesterone effects in the Wobbler mouse, a mutant presenting severe motoneuron degeneration and astrogliosis of the spinal cord. Implant of a single progesterone pellet (20 mg) during 15 days produced substantial changes in Wobbler mice spinal cord. Morphologically, motoneurons of untreated Wobbler mice showed severe vacuolation of intracellular organelles including mitochondria. In contrast, neuropathology was less pronounced in Wobbler mice receiving progesterone, together with a reduction of vacuolated cells and preservation of mitochondrial ultrastructure. Determination of mRNAs for the alpha 3 and beta 1 subunits of neuronal Na, K-ATPase, showed that mRNA levels in untreated mice were significantly reduced, whereas progesterone therapy re-established the expression of both subunits. Additionally, progesterone treatment of Wobbler mice attenuated the aberrant expression of the growth associated protein (GAP-43) mRNA which otherwise occurred in motoneurons of untreated animals. The hormone, however, was without effect on astrocytosis of Wobbler mice, determined by glial fibrillary acidic protein (GFAP) immunostaining. Lastly, progesterone treatment of Wobbler mice enhanced grip strength and prolonged survival at the end of the 15-day observation period. Recovery of morphology and molecular motoneuron parameters of Wobbler mice receiving progesterone, suggest a new and important role for this hormone in the prevention of spinal cord neurodegenerative disorders. PMID- 12650718 TI - Interactions of estrogen and insulin-like growth factor-I in the brain: molecular mechanisms and functional implications. AB - In the brain, as in other tissues, estradiol interacts with growth factors. One of the growth factors that is involved in the neural actions of estradiol is insulin-like growth factor-I (IGF-I). Estradiol and IGF-I cooperate in the central nervous system to regulate neuronal development, neural plasticity, neuroendocrine events and the response of neural tissue to injury. The precise molecular mechanisms involved in these interactions are still not well understood. In the central nervous system there is abundant co-expression of estrogen receptors (ERs) and IGF-I receptors (IGF-IRs) in the same cells. Furthermore, the expression of estrogen receptors and IGF-I receptors in the brain is cross-regulated. In addition, using specific antibodies for the phosphorylated forms of extracellular-signal regulated kinase (ERK) 1 and ERK2 and Akt/protein kinase B (Akt/PKB) it has been shown that estradiol affects IGF-I signaling pathways in the brain. Estradiol treatment results in a dose-dependent increase in the phosphorylation of ERK and Akt/PKB in the brain of adult ovariectomized rats. In addition, estradiol and IGF-I have a synergistic effects on the activation of Akt/PKB in the adult rat brain. These findings suggest that estrogen effects in the brain may be mediated in part by the activation of the signaling pathways of the IGF-I receptor. PMID- 12650719 TI - Evolution of oestrogen functions in vertebrates. AB - Steroidal oestrogens have been isolated from marine and terrestrial animals representative of all major classes of vertebrates including fish, amphibians, reptiles, birds and mammals. In general, oestrogens are responsible for most features characteristic of the female sex of a species, such as metabolic, behavioural and morphological changes during the stages of reproduction; they also support several processes in males. The evolution of the hormonal system always involves both the ligand and its sites of interaction. In the case of oestrogens, the steroid producing enzymes, mainly the aromatase complex, and the oestrogen receptor belong together within their co-evolution. The finding of oestrogenic steroids, the more recent identification of aromatase and receptor genes and their expression fit together, thereby confirming the importance for all vertebrates. Within the present paper, the evolution of the physiological functions of oestrogens from oviparous vertebrates to Eutherian mammals, oestrogen biosynthesis, metabolization and signalling pathways will be reviewed in detail. PMID- 12650720 TI - Stimulation of alkaline phosphatase activity in Ishikawa cells induced by various phytoestrogens and synthetic estrogens. AB - Xenoestrogens, phytoestrogens and synthetic estrogens, are able to bind to estrogen receptors, and to mimic estrogenic activities in a cell and tissue specific manner. For the characterization of environmental estrogens mainly mammary derived and yeast based models have been used. The aim of this study was therefore to assess selected natural and synthetic compounds in an endometrial derived model. We measured the relative estrogenic potency of phytoestrogens (genistein, daidzein, coumestrol, some naringenins), synthetic estrogens (bisphenol A, octylphenol, nonylphenol, o,p'-DDT), mycoestrogen (zearalanone) as well as extracts of Cimicifuga racemosa on alkaline phosphatase (AlkP) activity in the endometrial derived adenocarcinoma cell line Ishikawa. We used a modified multiwell plate in vitro bioassay based on the estrogen-specific and dose dependent enhancement of AlkP activity in this cell line. Estradiol, which induced AlkP at levels as low as 10(-8)M, was used as positive control. Most of the compounds studied showed a clear dose-dependent estrogenic effect. Compared to the vehicle control (ethanol) all phyto- and mycoestrogens, stimulated the AlkP activity 2-4-fold at a concentration of 10(-6)M. The synthetic chemicals bisphenol A and nonylphenol showed an effect at 10(-6)M, octylphenol at 10(-5)M. Effects of o,p'-DTT could not be measured. ICI 182,780, a pure estrogen receptor antagonist, significantly inhibited these effects. The latter result demonstrated the estrogen receptor dependency of this process. In summary, most of the phytoestrogens and industrial chemicals tested, behaved as estrogen receptor agonists in terms of the stimulation of AlkP activity. PMID- 12650721 TI - Endocrine disruptors: from Wingspread to environmental developmental biology. AB - The production and release of synthetic chemicals into the environment has been a hallmark of the "Second Industrial Revolution" and the "Green Revolution." Soon after the inception of these chemicals, anecdotal evidence began to emerge linking environmental contamination of rivers and lakes with a variety of developmental and reproductive abnormalities in wildlife species. The accumulation of evidence suggesting that these synthetic chemicals were detrimental to wildlife, and potentially humans, as a result of their hormonal activity, led to the proposal of the endocrine disruptor hypothesis at the 1991 Wingspread Conference. Since that time, experimental and epidemiological data have shown that exposure of the developing fetus or neonate to environmentally relevant concentrations of certain synthetic chemicals causes morphological, biochemical, physiological and behavioral anomalies in both vertebrate and invertebrate species. The ubiquitous use, and subsequent human exposure, of one particular chemical, the estrogen mimic bisphenol A (BPA), is the subject of this present review. We have highlighted this chemical since it provides an arresting model of how chemical exposure impacts developmental processes involved in the morphogenesis of tissues and organs, including those of the male and female reproductive systems, the mammary glands and the brain. PMID- 12650722 TI - Prohormones and sport. AB - Several precursors of testosterone and nandrolone introduced on the nutritional supplement market as performance enhancing drugs are banned in sports. Until now they are legally sold without a prescription in the US. Results of excretion studies with related compounds including 7-keto-DHEA and 1-androstenediol are presented. The main metabolites of 7-keto-DHEA are 7-hydroxylated compounds. The commercial 1-androstenediol preparation was contaminated with several other anabolic steroids. Oxidation of 1-androstenediol to 1-androstenedione seems to be the major renal metabolic pathway. Additionally contaminated nutritional supplements containing banned substances not indicated on the label were administered. The results of the excretion studies indicate that after the intake of amounts substantially lower than the recommended dose athletes can fail a doping test for periods up to 120 h. PMID- 12650723 TI - Reproduction gone awry: medical anthropological perspectives. PMID- 12650724 TI - Global infertility and the globalization of new reproductive technologies: illustrations from Egypt. AB - Infertility is a problem of global proportions, affecting on average 8-12 percent of couples worldwide. In some societies, however-particularly those in the "infertility belt" of sub-Saharan Africa-as many as one-third of all couples are unable to conceive. Factors causing high rates of tubal infertility in parts of the developing world include sexually transmitted, postpartum, and postabortion infections; however, male infertility, which is rarely acknowledged, contributes to more than half of all cases. Unfortunately, the new reproductive technologies (NRTs) such as in vitro fertilization (IVF), which are prohibitively expensive and difficult to implement in many parts of the developing world, represent the only solution to most cases of tubal and male infertility. Not surprisingly, these technologies are rapidly globalizing to pronatalist developing societies, where children are highly desired, parenthood is culturally mandatory, and childlessness socially unacceptable. Using Egypt as an illustrative case study, this paper examines five of the major forces fueling the global demand for NRTs; these include demographic and epidemiological factors, the fertility-infertility dialectic, problems in health care seeking, gendered suffering, and adoption restrictions. Following this overview, a detailed examination of the implications of the rapid global spread of NRTs to the developing world will be offered. By focusing on Egypt, where nearly 40 IVF centers are in operation, this article will demonstrate the considerable constraints on the practice and utilization of NRTs in a developing country on the "receiving end" of global reproductive technology transfer. The article concludes by stressing the need for primary prevention of infections leading to infertility, thereby reducing global reliance on NRTs. PMID- 12650725 TI - With or against nature? IVF, gender and reproductive agency in Athens, Greece. AB - Based on ethnographic research in Athens, this paper argues that in vitro fertilization (IVF) in urban Greece does not so much make explicit the social construction of nature, as has been argued of the US and UK, but is accommodated into a prior understanding of "nature" as socially realized. Calling upon an ethic of maternal sacrifice, Athenian women see themselves taking charge of a natural process, often correcting damage done to them by nature. A sense that adults should produce children in order to realize their natures and be completed as women and men also poses particular, and gendered, ethical questions of fertility technologies: is this a proper way of realizing nature? While those who successfully use IVF depict assisted conception as "a natural" and "proper way of reproduction," some fear that others will view their child as abnormal. Contests over "normal" reproduction are articulated to a tradition/modernity dichotomy which Athenians discuss through an idiom of maturity. Believing that Greek society is not always mature enough to understand how natural IVF really is, couples turning to IVF follow one of the two strategies: to educate others or to completely hide it. Both groups want to guard against a view of IVF-in the terms of this volume-as a kind of "reproduction gone awry." PMID- 12650726 TI - Why adoption is not an option in India: the visibility of infertility, the secrecy of donor insemination, and other cultural complexities. AB - Child adoption in the face of reproduction gone awry continues to remain an under researched aspect of contemporary Indian reality. This paper seeks to unpack some of the critical cultural issues underscoring the deep-seated reluctance towards adoption. Drawing on a larger multi-sited research project examining the experience of infertility and assisted conception in India, the paper sheds light on the state of current adoption practices in India. Thus, when faced with infertility, couples in this research emerged as favouring secret gamete donation as a means of bypassing infertility rather than the option of adoption. Invoking the concept of systematic misrecognition, the paper situates the modalities of salvaging infertility, either through medically assisted conception or adoption, as structuring infertile people's quest for children. The paper relates the perceived stigma associated with infertility treatment and adoption with the inclusion of a "third party" that fractures the culturally conceptualized boundaries of family as inextricably tied to the conjugal bond. It is therefore argued that secrecy is born out of a need to obfuscate a "public and visible" violation of a culturally priced ideal that views an intimate connection between the "married body" and the progeny. Adoption continues to remain an undesirable option because the links between an adopted child and the social parent become a public, vocal, and visible admission of infertility that cannot be subsumed, like donated gamete conception, under a conspiracy of silence. PMID- 12650727 TI - Unhappy endings: a feminist reappraisal of the women's health movement from the vantage of pregnancy loss. AB - This essay contrasts the rosy birth scenarios of the natural childbirth movement with reproductive disaster stories of members of pregnancy loss support groups and women from toxically assaulted communities in the US who have suffered pregnancy loss. I argue that both biomedical obstetrics and the women's health movement critique of it share a belief in the ability to control reproduction so that there will be a positive outcome. I show that this emphasis on happy endings (whether believed to be the result of medical intervention, or women's natural inborn powers to reproduce) exacerbates the experience of those whose pregnancies do not end happily. I show how the women's health movement's emphasis on the importance of women being in control of their own bodies is related to a broader "culture of meritocracy" which contributes to maternal blame (and self-blame) when pregnancies are not perfect. PMID- 12650728 TI - Burning bridges: policy, practice, and the destruction of midwifery in rural Costa Rica. AB - The trend toward hospitalization of birth has a long history in Costa Rica and currently approximately 98% of births take place in the clinical setting. Impoverished rural areas, like the town of Buenos Aires, lag behind national trends and only recently has birth moved from the home to the hospital. Costa Rica's midwife certification program co-opted rural midwives as bridges to biomedicalization, responsible for both pushing women into the biomedical setting and filling the gaps left by a limited national health care system. Despite the eventual illegalization of key practices and of home birth itself, local use of midwives' services continues, albeit with local demands that have transformed midwives into bridges to biomedical care in ways unanticipated by and invisible to national programmers. Midwives provide key services like prenatal massage, treatment of pregnancy crises, and attending unforeseen home births and women unable to afford the modest costs of hospitalization. Yet, midwives report increasing dissatisfaction and the desire to stop providing services in their communities. Practices like prenatal massage are in demand, but are no longer embedded in a system of local exchange that is socially and economically meaningful. Midwives blame their clientele for their dissatisfaction, but directly link these changes to the notions of professionalism, compensation, and changing community values. Thus, the social relationship between midwives and their clients must also be understood as a destructive force burning midwifery as a bridge to safe birth. In this essay, I argue that the process of both remodeling and subsequently destroying midwifery practices begun in the formal health care sector at the national level continues at the local level through changing values and meanings associated with midwives' practices. PMID- 12650729 TI - Home-birth emergencies in the US and Mexico: the trouble with transport. AB - Proponents of the global Safe Motherhood Initiative stress that primary keys to safe home birth include transport to the hospital in cases of need and effective care on arrival. In this article, which is based on interviews with American direct-entry midwives and Mexican traditional midwives, I examine what happens when transport occurs, how the outcomes of prior transports affect future decision-making, and how the lessons derived from the transport experiences of birthing women and midwives in the US and Mexico could be translated into improvements in maternity care. My focus is on home birth in urban areas in Mexico and the US. In both countries, biomedicine and home-birth midwifery exist in separate cultural domains and are based on distinctively different knowledge systems. When a midwife transports a client to the hospital, she brings specific prior knowledge that can be vital to the mother's successful treatment by the hospital system. But the culture of biomedicine in general tends not to understand or recognize as valid the knowledge of midwifery. The tensions and dysfunctions that often result are displayed in midwives' transport stories, which I identify as a narrative genre and analyze to show how reproduction can go unnecessarily awry when domains of knowledge conflict and existing power structures ensure that only one kind of knowledge counts. This article describes: (1) disarticulations that occur when there is no correspondence of information or action between the midwife and the hospital staff; and (2) fractured articulations of biomedical and midwifery knowledge systems that result from partial and incomplete correspondences. These two kinds of disjuncture are contrasted with the smooth articulation of systems that results when mutual accommodation characterizes the interactions between midwife and medical personnel. The conclusion links these American and Mexican transport stories to their international context, describing how they index crosscultural markers, and suggest solutions, for "the trouble with transport." PMID- 12650730 TI - Genetic counseling gone awry: miscommunication between prenatal genetic service providers and Mexican-origin clients. AB - Amniocentesis, and other prenatal genetic tests, have become a well-established feature of modern prenatal care. But these tests place a considerable decision making burden on the expectant mothers to whom they are offered: the genetic issues involved are complex and the appropriate course of action sometimes ambiguous. Genetic counseling aims to help pregnant clients make an informed decision about prenatal genetic tests. But the clientele of prenatal genetic counseling has changed significantly in the years since the practice was established. Clients were once a self-selected group of women well-informed about the genetic services being offered. In contrast, clients now include an increasing number of women, particularly ethnic minority women, who had no prior knowledge of genetic testing, but were found to be at risk of birth defects after routine screening. Little is known about how well genetic counseling serves the needs of this new clientele. This paper investigates the possibility that miscommunication between genetic counselors and their Mexican-origin clients contributed to the higher rates of amniocentesis refusal. We interviewed 156 pregnant Mexican-origin women who screened positive on a blood test routinely offered in California to detect birth defects. We also observed the genetics consultations of a sub-sample of the women. We identified five common sources of miscommunication: (1) Medical jargon; (2) The non-directive nature of counseling; (3) The inhibitions of counselors stemming from misplaced cultural sensitivity; (4) Problems of translation; (5) Problems of trust. We found that many Mexican origin women are skeptical of genetic testing and do not easily surrender their own lay theories about the causes of their condition. In order to dislodge the misunderstandings of their clients, counselors must give clients the opportunity to air their own views, however contrary to those of genetics professionals these may be. PMID- 12650731 TI - Emplotting children's lives: developmental delay vs. disability. AB - While it is increasingly possible to envision "perfect" babies, it is not always the case that reproduction actually proceeds according to individual will; for example, there has been no recent reduction in rates of childhood disability. Nevertheless, in most studies of new reproductive technologies, the birth of those children whom few would actively choose-"defective" or disabled infants-is presented only in hypothetical terms. This paper argues for expanding the domain of reproduction to include research on the parenting of children with disabilities. Based on a qualitative research project carried out at a hospital based newborn follow-up program that serves as an evaluation site determining eligibility for early intervention services for infants and young children with disabilities, this paper focuses on a particular part of women's experience of acquiring new knowledge about personhood and disability, that is, on the period of time when a woman has recently had confirmed that reproduction has, in her case, gone awry. Disability in many cultures, including the United States, diminishes personhood. I suggest that American mothers' narratives, by utilizing the concept of developmental delay, can assert personhood, or rather, the potential for its future attainment; in doing so, they justify ongoing nurturance of a disabled child in spite of negative attitudes about disability. A particular case of one mother's emplotment of her child's life within a story of developmental delay, in competition with the physician's story of disability, is analyzed. The paper concludes with reflections on how stories of developmental delay told by mothers just encountering a diagnosis of disability may differ from the stories told by those who have experienced mothering a disabled child over time, and on the implications of these differences for the cultural construction of personhood in the United States. PMID- 12650732 TI - Polygamy, disrupted reproduction, and the state: Malian migrants in Paris, France. AB - For Malian women, migration from West Africa to France has disrupted widely shared understandings of marriage and reproduction. In response to restrictive immigration policies, men and women routinely confront the challenges of polygamy, public disapproval of high fertility, and biomedical messages promoting contraception. Although many Malians continue to be strongly pronatalist, within a family, spouses may experience contradictory pressures and objectives regarding reproduction, particularly in polygamous marriages. Because women are more likely than men to interact with nurses and doctors in the context of maternity and child health care visits, they are systematically confronted by encouragement to contracept. French population policy is contradictory in this regard, as it has been strongly pronatalist throughout the 20th century, yet is equally strongly anti-natalist with regard to immigrant populations. Recent anti-immigrant policies such as the Pasqua law prohibiting polygamy have emerged as influences shaping men's and women's contested reproductive goals. Men tend to oppose contraception, citing Islamic doctrine while women increasingly justify contraceptive use in response to government policies and biomedical encouragement. In contrast, polygamy also may generate pregnancy rivalries as wives strategize to enhance their reproductive careers and thus to retain immigrant status. PMID- 12650733 TI - Antiabortion positions and young women's life plans in contemporary Ireland. AB - At a critical time when Ireland's abortion ban faces legal challenges and the number of women obtaining abortions abroad each year continues to climb, some antiabortion advocates have turned their attention toward the social factors that influence women's abortion decision-making. Through an analysis of articles carried in the Irish mainstream and Catholic presses, this article examines how antiabortion advocates since the late 1990s have promoted an "antiabortion, pro motherhood" message in response to trends that they identify as indicating that Irish reproduction has "gone awry". Antiabortion activists have focused in particular on the life plans of young, middle-class, career-oriented women, many of whom have benefited from increased employment opportunities within Ireland. These women are more likely than young women in past generations to postpone childbearing or opt for abortion in the face of an unwanted pregnancy, and thus, symbolize for antiabortion advocates the devaluation of a "traditional" Irish culture centered on the privileging of motherhood and married family life. This article examines antiabortion ideologies deployed around motherhood, work, and childcare, and argues that antiabortion advocates' "pro-motherhood" campaign fails to adequately respond to the changing realities of young, middle-class Irish women's life opportunities and expectations. PMID- 12650734 TI - Post-diagnostic abortion in Germany: reproduction gone awry, again? AB - Routine use of prenatal diagnostic technologies (PDTs) such as ultrasound and amniocentesis result in the detection of a small percentage of fetal anomalies. For those women faced with the diagnosis of fetal disability, a decision must be made to continue or terminate the pregnancy. When the diagnosis is merely hypothetical, the discursive specter of post-diagnostic abortion is shaped by social and historical contexts in which interested discourses (regional, political, ethical, and religious) weigh in with varying degrees of authority and influence. However, when the diagnosis is actual, in this sample population of women, an estimated minimum of 90% opt to terminate their pregnancies. Data collected at two German hospitals-one in former East Germany, one in former West Germany-illuminate rates of PDT use and provide data with which to discuss the specter of post-diagnostic abortion in relation to mainstream medical discourses, Germany's divided history, abortion politics, feminism, disability activism, and religion. These data demonstrate how reproductive discourses are shaped by ideological and historical contingencies, even when women's ultimate reproductive decisions are not. PMID- 12650738 TI - abcb1ab P-glycoprotein is involved in the uptake of citalopram and trimipramine into the brain of mice. AB - The phenomenon of a heterogeneous response to the same drug in different patients is well-known. An important reason is that, even at equal concentrations, the bioavailability of a drug depends on the interaction of the drug with the blood brain barrier (BBB). In part, this is due to the drug-transporting P-glycoprotein (P-gp), a product of the multiple drug resistance gene (ABCB1), which can transport drugs against a concentration gradient across the BBB back into the plasma and thereby reduce the bioavailability in the brain. In the present study, we have examined the uptake of the antidepressants citalopram and trimipramine into the brain of abcb1ab knockout mice compared with controls. One hour after s.c. injection of the drugs, concentrations of the two drugs and of the metabolite d-trimipramine in brain, spleen, kidney, liver and plasma were measured with HPLC. Significantly higher brain concentrations in knockout mice, showing that these drugs are substrates of P-gp and that the presence of P-gp reduces the effective bioavailability of these substances in the brain. The results of our study contradict an earlier report that citalopram is not actively transported from endothelial cells. These results were derived from an in vitro study, showing that due to the complexity of the BBB-drug interaction, it is difficult to transfer results from in vitro studies to the in vivo situation. We hypothesize that inter-individual differences in the activity of the ABCB1 gene can account in part for the great variation in clinical response to antidepressants in psychiatric patients, even at comparable plasma levels. PMID- 12650739 TI - Comparing anxiety disorders and anxiety-related traits in bipolar disorder and unipolar depression. AB - The frequent comorbidity of anxiety disorders and mood disorders has been documented in previous studies. However, it remains unclear whether specific anxiety traits or disorders are more closely associated with unipolar major depression (MDD) or bipolar disorder (BPD). We sought to examine whether MDD and BPD can be distinguished by their association with specific types of anxiety comorbidity. Individuals with a primary lifetime diagnosis of either bipolar disorder (N=122) or major depressive disorder (N=114) received diagnostic assessments of anxiety disorder comorbidity, and completed questionnaires assessing anxiety sensitivity and neuroticism. The differential association of these anxiety phenotypes with MDD versus BPD was examined with multivariate modeling. Panic disorder and generalized anxiety disorder (GAD) specifically emerged amongst all the anxiety disorders as significantly more common in patients with BPD than MDD. After controlling for current mood state, anxiety sensitivity and neuroticism did not differ by mood disorder type. This study supports prior research suggesting a specific panic disorder-bipolar disorder connection, and suggests GAD may also be differentially associated with BPD. Further research is needed to clarify the etiologic basis of anxiety disorder/BPD comorbidity and to optimize treatment strategies for patients with these co occurring disorders. PMID- 12650740 TI - Epidemiology, implications and mechanisms underlying drug-induced weight gain in psychiatric patients. AB - Body weight gain frequently occurs during drug treatment of psychiatric disorders and is often accompanied by increased appetite or food craving. While occurrence and time course of this side effect are difficult to predict, it ultimately results in obesity and the morbidity associated therewith in a substantial part of patients, often causing them to discontinue treatment even if it is effective. This paper reviews the available epidemiological data on the frequency and extent of weight gain associated with antidepressant, mood-stabilizing, and antipsychotic treatment. Possible underlying pathomechanisms are discussed with special attention to central nervous control of appetite including the role of leptin and the tumor necrosis factor system. Metabolic alterations induced by drug treatment such as type 2 diabetes mellitus and the metabolic syndrome are also considered. Weight gain appears to be most prominent in patients treated with some of the second generation antipsychotic drugs and with some mood stabilizers. Marked weight gain also frequently occurs during treatment with most tricyclic antidepressants, while conventional antipsychotics typically induce only slight to moderate weight gain. Serotonin reuptake inhibitors may induce weight loss during the first few weeks, but some of them induce weight gain during long-term treatment. Several antidepressant and antipsychotic drugs are identified which reliably do not cause weight gain or even reduce weight. Based on these insights, countermeasures to manage drug-induced weight gain are suggested. PMID- 12650741 TI - Increased beclomethasone-induced vasoconstriction in women with posttraumatic stress disorder. AB - It has been hypothesized that patients with posttraumatic stress disorder (PTSD) show increased glucocorticoid sensitivity. The study tested beclomethasone induced vasoconstriction (BIV), a measure of peripheral glucocorticoid sensitivity, in women with PTSD. A case-control design was employed in 33 PTSD patients and 33 healthy controls. BIV was tested using beclomethasone dipropionate (1-100 micro g/ml). Vasoconstriction was assessed after 15-18 h. Waking and afternoon salivary cortisol concentrations were measured. BIV ratings were significantly increased in PTSD at beclomethasone concentrations from 10-100 micro g/ml. Salivary cortisol concentrations did not differ between groups or correlate with BIV. Preliminary evidence has been found for increased peripheral glucocorticoid sensitivity in PTSD. Further study is required to replicate this finding and assess its relationship to the pathophysiology of the disorder. PMID- 12650742 TI - Plasma levels of soluble transferrin receptors and Clara cell protein (CC16) during bipolar mania and subsequent remission. AB - Clara cell protein (CC16) and transferrin receptor (TfR) have been reported as possible biological markers for major depression and schizophrenia. However, the alternations of plasma TfR and CC16 levels and the influences of numerous clinical variables on them during bipolar mania are not sufficiently described. We investigated the immune function of 36 bipolar I, manic (DSM-IV) patients with Young Mania Rating Scale (YMRS) scores > or =26 as well as during the subsequent remission (YMRS < or =12) and age- and sex- matched healthy controls. The plasma TfR levels were increased during acute mania along with subsequent remission and were independent of medication status, individual variations, clinical and erythrocyte variables. Among inflammatory parameters and haematological variables, the plasma TfR levels merely had significant and negative relationship with the percentage of monocyte in circulating leukocyte counts despite of elevated plasma soluble interleukin-2 receptors levels during bipolar mania. The plasma levels of CC16 of bipolar patients did not significantly alter during acute mania, whereas smoking, body mass index, and co-existing psychotic features collectively contributed 42% of the plasma levels of CC16. We provide additional evidence to indicate the pathophysiological role of the immune systems in affective disorders. It is suggested that the elevation of plasma TfR levels might be a trait phenomenon in bipolar disorder. PMID- 12650743 TI - Potential use of the topical niacin skin test in early psychosis -- a combined approach using optical reflection spectroscopy and a descriptive rating scale. AB - The niacin skin phenomenon reflects a prostaglandin (PG) mediated flush and oedema reaction. As PG metabolism is linked to breakdown of membrane lipids, diminished sensitivity to niacin application suggests potential disturbance in membrane phospholipid-arachidonic acid-PG pathways. We aimed to evaluate and quantify topical niacin skin reaction in early psychosis using optical reflection spectroscopy (ORS) and a new descriptive assessment scale integrating time course, redness, and oedema. Niacin skin tests were performed on 25 medicated first-episode psychosis patients fulfilling DSM-IV criteria for schizophreniform psychosis or schizophrenia and on 25 healthy controls. Nicotinic acid was applied in four dilution steps to the subjects inner forearm skin and skin reaction was consecutively assessed using ORS and a seven point rating scale. Both descriptive ratings and spectroscopic measures revealed significant group differences at the lower niacin concentrations (0.001 and 0.0001 M). At higher concentrations (0.01 and 0.1 M) only descriptive ratings were capable to show significant group effects. Data of both methods showed moderate to strong correlation (r=0.605) as long as the erythema was not affected by the oedema. The data suggest that niacin sensitivity is inversely correlated with negative symptoms. Both methods demonstrate that niacin sensitivity is impaired in a group of first episode psychosis patients and are therefore able to distinguish a subgroup of patients with metabolic impairment. Niacin sensitivity in high risk populations and the specificity of impaired skin response are subjects of further investigation. PMID- 12650744 TI - Tail-suspension induced hyperthermia: a new measure of stress reactivity. AB - The tail suspension test (TST), an antidepressant screening paradigm, uses the uncontrollable, inescapable stressor of tail suspension to elicit immobility. As hyperthermia occurs following numerous stressors, hyperthermia might exist following the TST. We tested whether tail suspension induced hyperthermia (TSIH) was a distinct variable for TST. Hyperthermia was measured by two methods: a rectal probe and a subcutaneously implanted microchip (ELAMS()). In outbred ICR male mice, TSIH was robustly demonstrated compared to control (No-TST) mice. TSIH peaked after TST and remained elevated at 120 min. Among five (129/SvEvTac, A/J, C57BL/6J, NMRI and ICR) strains examined for TSIH, significant strain variations were detected. NMRI showed the highest temperature rise (2.3 degrees C) and A/J mice showed the lowest (0.6 degrees C). Sex differences were found for the C57BL/6J and NMRI strains on TSIH. TSIH and duration of immobility were not significantly correlated (r=0.22, P=0.17) in outbred mice. Both duration of TST immobility and TSIH were measured when ICR male mice were administered diazepam, imipramine (a TCA antidepressant), venlafaxine (a SNRI antidepressant), sertraline and paroxetine (SSRI antidepressants), propranolol and nadolol (beta adrenergic receptor blockers), CP-154,526 (a CRF(1) receptor antagonist), and indomethacin (a cyclo-oxygenase inhibitor). Diazepam dose-dependently increased immobility and decreased TSIH. Propranolol blocked TSIH, but nadolol had no effect. Antidepressants showed more complex patterns of effects with venlafaxine, sertraline, and paroxetine inhibiting TSIH. TSIH demonstrated inter-strain variability, sex differences and a distinct pharmacology, suggesting that TSIH provides an independent, robust physiologic parameter to supplement the TST paradigm. This TSIH method may prove useful for pharmacologic, transgenic, and mechanistic studies. PMID- 12650745 TI - Oral citalopram and reboxetine challenge tests before and after selective antidepressant treatment. PMID- 12650748 TI - Rapid and simple method to determine morphine and its metabolites in rat plasma by liquid chromatography-mass spectrometry. AB - A rapid and simple method for the determination of morphine (M), normorphine (NM), morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) in plasma by high-performance liquid chromatographic separation with mass spectrometric detection (HPLC-MS) has been developed. Samples (40 microl) were cleaned-up by protein precipitation with two volumes (80 microl) of acetonitrile and reconstituted in formic acid 0.1% in water. Naloxone was used as internal standard. Analytes were separated on a phenyl-hexyl column using a step-gradient (1 ml/min) of acetonitrile and formic acid in water. Acetonitrile was added post column (0.3 ml/min). Quantification of morphine and its metabolites was achieved with an Agilent 1100 series HPLC-MS system equipped with electrospray interface set to selected ion-monitoring (SIM) mode. Calibration curves covered a wide range of concentrations (2.44-10,000 nM) and were best fitted with a weighed quadratic equation. The limits of quantification achieved with this method were 2.44 nM for M and 4.88 nM for NM, M3G and M6G. The method proved accurate (85 98%), precise (C.V.<10%) and was successfully applied to a wide range of in vitro and in vivo pharmacokinetic studies in rodents. PMID- 12650747 TI - Factorial design for the development of automated solid-phase extraction in the 96-well format for determination of tesaglitazar, in plasma, by liquid chromatography-mass spectrometry. AB - An analytical method was developed for the determination, in blood plasma, of a novel peroxisome proliferator-activated receptor (PPAR) agonist drug, tesaglitazar. The drug and the isotope labelled internal standard were isolated by solid-phase extraction (SPE) on hexylsilica, separated by reversed-phase liquid chromatography and quantified by tandem mass spectrometry. Factorial design and a robotic sample processor were employed in the exploration and optimisation of the SPE procedure in the 96-well format. This allowed rapid development of the method, notably limiting the process to four experiments before validation. The detectability was greatly improved by utilising the formation of sodium adducts in atmospheric pressure positive ionisation mass spectrometry. Absolute recovery was more than 95% with a coefficient of variation of 5% at a level of 8.7 nM. The accuracy and precision of the automated SPE method presented here matched the excellence of the previously used method based on manual liquid-liquid extraction. Furthermore, the method resulted in an increased sample throughput. PMID- 12650749 TI - Complementary use of ion trap/time-of-flight mass spectrometry in combination with capillary high-pressure liquid chromatography: early characterization of in vivo metabolites of the cathepsin K inhibitor NVP-AAV490 in rat. AB - Cathepsin K is a cysteine proteinase, primarily expressed in osteoclasts, which has a strong collagenolytic activity and plays an essential role involved in bone matrix degradation. Its inhibition could provide a novel approach to the treatment and prevention of osteoporosis. One structural class of lead compounds in our cathepsin K inhibitors program is based on an arylaminoethyl amide scaffold, which has potential metabolic weak points that might be stabilized by appropriate chemical modification(s). For the identification of potential metabolic "soft spots" and the rational design of improved derivatives, early biotransformation of a potent arylaminoethyl amide cathepsin K inhibitor (NVP AAV490-NX) was investigated in plasma, urine and liver homogenates of rats after intravenous bolus administration of 10 mg/kg. The detection and identification of metabolites was achieved by high-resolution mass spectrometry (time-of-flight MS) and multi-dimensional mass spectrometry (ion trap MS). Both mass spectrometers were combined with reversed-phase capillary high-performance liquid chromatography columns. It was demonstrated that both mass analyzers complement each other and that, even in the sub-nanogram range, the resulting set of MS data can be successfully used to elucidate most of the metabolic changes unambiguously, solely by mass spectrometric techniques. The proposed metabolite structures were additionally corroborated by exact mass measurement of the protonated molecular ions to confirm the predicted elemental composition, by determination of the number of the exchangeable hydrogen atoms replacing water against deuterium oxide as mobile phase and, in one case, by an MS(3) product ion experiment in order to elucidate the site of conjugation. PMID- 12650750 TI - Clinical use of polyethylene glycols as marker substances and determination in urine by liquid chromatography. AB - Adulteration of samples is a serious problem in the analysis of drugs of abuse. One of the most frequent methods is substitution of urines by "clean" urines to gain false-negative results in laboratory tests for drugs of abuse. One way to approach this problem may be to label the patient's urine with marker substances which are given orally prior to the delivery of urine. This concept is based on methods for determining malabsorption in pediatric medicine. We report a protocol for evaluating low-molecular-mass polyethylene glycols as enteral labelling marker substances. For monitoring renal excretion of the ingested polyethylene glycols we have developed and optimised an isocratic reversed-phase high performance liquid chromatographic method with automatic sample cleanup by column switching in the back-flush technique and with RI detection. The chromatographic procedure is simple, reliable and rapid, allowing a high sample throughput for routine screening. PMID- 12650751 TI - Liquid chromatographic assay for the simultaneous determination of indole-3 carbinol and its acid condensation products in plasma. AB - A high-performance liquid chromatographic method was developed for the simultaneous determination of indole-3-carbinol (I3C), 3,3'-diindolylmethane (DIM), [2-(indol-3-ylmethyl)-indol-3-yl]indol-3-ylmethane (LTr(1)), and indolo[3,2b]carbazole (ICZ). Compounds were extracted from mouse plasma using tert.-butyl methyl ether, incorporating 4-methoxy-indole as internal standard. Chromatographic separation utilized a Waters Symmetry RP18 in tandem with a Thermoquest BDS C(18) column, an acetonitrile-water gradient and UV (280 nm) in series with fluorescence (ex. 335 nm; em. 415 nm) detection. Calibration curves were linear (r(2)>0.99) between 50 and 15,000 ng/ml for I3C; 150 and 15,000 ng/ml for LTr(1); and 0.15 and 37.5 ng/ml for ICZ and the method was reproducible and precise (within-day and between-day coefficients of variation below 9.7 and 13%, respectively). The method described is suitable for comprehensive pharmacokinetic studies with indole-3-carbinol. PMID- 12650752 TI - Non-linear heteroscedastic regression model for determination of methotrexate in human plasma by high-performance liquid chromatography. AB - Generalized least squares regression with variance function estimation was used to derive the calibration function for measurement of methotrexate plasma concentration and its results were compared with weighted least squares regression by usual weight factors and also with that of ordinary least squares method. In the calibration curve range of 0.05 to 100 microM, both heteroscedasticity and non-linearity were present therefore ordinary least squares linear regression methods could result in large errors in the calculation of methotrexate concentration. Generalized least squares regression with variance function estimation worked better than both the weighted regression with the usual weight factors and ordinary least squares regression and gave better estimates for methotrexate concentration. PMID- 12650753 TI - Simple immunoaffinity method to purify recombinant hepatitis B surface antigen secreted by transfected mammalian cells. AB - Purification of recombinant hepatitis B surface antigen (recHBsAg) produced in a stable Chinese hamster ovary (CHO) cell line was evaluated using Linx Affinity Purification System (Invitrogen, USA). To purify HBsAg secreted by this cell line, a murine monoclonal antibody (MAbAH1) raised against native HBsAg was used. The purified AH1MAb was conjugated with phenyldiboronic acid (PDBA) and immobilized on the immunoaffinity chromatographic support. Using an optimized protocol the affinity column was able to purify recHBsAg from supernatant of mammalian cells cultures with more than 80% purity. This method showed to be simple and quicker than the current ultracentrifugation methods. The method is also efficient and economical in obtaining purified recHBsAg. PMID- 12650754 TI - Development of a liquid chromatography-mass spectrometric method for measuring the binding of memantine to different melanins. AB - A sensitive and selective liquid chromatography-mass spectrometric method was validated for the determination of free memantine in melanin binding studies. The sources of melanin studied were sepia, synthetic and bovine melanin. Memantine was chromatographed on a reversed-phase column (Prodigy 5 microm, ODS(3), 100 A, 100 x 4.6 mm) using gradient elution with mobile phases of 0.1% formic acid in deionised water and 0.1% formic acid in methanol at a flow-rate of 0.8 ml/min. The mode of ionisation was atmospheric pressure-electrospray and detection by single ion monitoring of the memantine ion m/z 180. Validation of the method showed that the assay was linear from 0.1 to 1200 nM and 0.5 to 1200 nM memantine in deionised water and phosphate-buffered saline (PBS), respectively. Accuracy for sample preparations in deionised water was between 80 and 108% and between 80 and 123% for PBS. For both media, intra- and inter-day precision was below 1% for retention time and below 5% for analyte peak area. At the LLOQ, the variation of peak area was less than 17%. Binding of memantine to melanin was measured indirectly by determining the unbound fraction of memantine. After incubation of melanin with memantine, the sample was centrifuged and filtered to separate the memantine-melanin complex effectively from suspension. The filtrate was then assayed for free memantine from which the extent of binding was then calculated. PMID- 12650755 TI - On-line determination of fluconazole in blood and dermal rat microdialysates by microbore high-performance liquid chromatography. AB - To study the distribution of fluconazole in the dermis of the rat, on-line microdialysis using double-site sampling coupled with a microbore HPLC system was developed. The chromatographic conditions consisted of a mobile phase of 20 mM diammonium phosphate-acetonitrile (75:25, v/v, pH 7.0) pumped through a microbore C(18) column at 40 microl/min. The eluent was monitored with UV detector with UZ flow cell (30 mm path length) at 210 nm. A microbore 10-port pneumatic valve fitted with two loops of 1 microl was used to collect and directly inject microdialysates from jugular and dermal probes. The retention time was 5.8 min for fluconazole and 10.1 min for its fluorinated analog, UK-54373 used as a retrodialysis marker. The assay was precise, with inter- and intra-assay relative standard deviation values of 0.64 and 0.71%, respectively, and with a good linearity (r=0.999) in the range of 0.15-20 microg/ml with only 1 microl injected onto the column. The LOD and LOQ values for fluconazole were 0.100 and 0.150 microg/ml, respectively. The applicability of the method was demonstrated by studying the disposition of fluconazole in blood and dermis following i.v. bolus at a dose of 10 mg/kg. PMID- 12650756 TI - Development and validation of a liquid chromatography-tandem mass spectrometric assay for Eplerenone and its hydrolyzed metabolite in human plasma. AB - A sensitive and specific liquid chromatography-tandem mass spectrometry assay was developed to quantify the first selective aldosterone blocker Eplerenone (I) and its hydrolyzed metabolite (II) in human plasma. The analytes (I, II) and their stable isotope-labeled analogues as internal standards were extracted on a C(18) solid-phase extraction cartridge using a Zymark RapidTrace automation system. The chromatographic separation was carried out on a narrow-bore reversed-phase Zorbax XDB-C(8) HPLC column with a mobile phase of acetonitrile/water (40:60, v/v) containing 10 mM ammonium acetate (pH 7.4). The analytes were ionized using negative-to-positive switch electrospray mass spectrometry, then detected by multiple reaction monitoring with a tandem mass spectrometer. The precursor to product ion transitions of m/z 415-->163 and m/z 431-->337 was used to measure I and II, respectively. The assay exhibited a linear dynamic range of 10-2500 ng/ml of plasma for both I and II. The lower limit of quantification was 10 ng/ml for I and II. Acceptable precision and accuracy were obtained for concentrations over the standard curve ranges. A throughput of 80 human plasma standards and samples per run was achieved with run time of 5 min for each injection. The assay has been successfully used in analyses of human plasma samples to support clinical studies. PMID- 12650757 TI - Determination of Chinese hamster ovary cell-derived recombinant thyrotropin by reversed-phase liquid chromatography. AB - A reversed-phase high-performance liquid chromatography (RP-HPLC) methodology for the qualitative and quantitative analysis of human thyrotropin (hTSH) in CHO cell conditioned medium and in purified preparations has been set up and validated for accuracy, precision and sensitivity. A recovery test indicated a bias of less than 2% and intra-day and inter-day quantitative determinations presented relative standard deviations (RSD) always <7%, while sensitivity was 0.2 microg (RSD=5.6%). The novel methodology was applied to the study of the best cultivation conditions and was able to detect a significant difference in retention time (t(R)) between pituitary and recombinant hTSH, probably reflecting the influence of the heterogeneity of the carbohydrate moiety on the hydrophobic properties of the molecule. PMID- 12650758 TI - Purification strategy for recombinant Phl p 6 is applicable to the natural allergen and yields biochemically and immunologically comparable preparations. AB - The recombinant major grass pollen allergen Phl p 6 has been expressed with a N terminal 6 x His-tag sequence and subsequently purified using nickel-chelating Sepharose. After cleavage of the tag-sequence, a second pass over the affinity chromatography revealed that even untagged rPhl p 6 bound tightly. In order to determine if that property is typical for Phl p 6, the natural allergen was purified in the same way starting with a grass pollen extract. Indeed, nPhl p 6 could be highly enriched in one step using nickel-chelating Sepharose. In addition to this new powerful purification method, the results provide further information in that the recombinant and natural allergens share a lot of properties, since biochemical characteristics are reflected in the purification strategies. The preparations of natural and recombinant Phl p 6 were used for comparative electrophoretic, chromatographic and immunological analysis which demonstrated high similarity. PMID- 12650759 TI - Simultaneous determination of hydroxycinnamates and catechins in human urine samples by column switching liquid chromatography coupled to atmospheric pressure chemical ionization mass spectrometry. AB - A quantitative liquid chromatography mass spectrometry (LC-MS) methodology with online sample clean up by column switching is described for the simultaneous determination of the hydroxycinnamates, caffeic acid and chlorogenic acid, and of the catechins, epicatechin and catechin in human urine samples. Enzymatically treated urine samples were directly injected onto the LC-MS system, where sample clean up was performed by a reversed-phase Zorbax 300SB C(3) column and selective elution of the target compounds onto a Zorbax SB C(18) column resulted in final separation prior to detection by atmospheric pressure chemical ionization (APCI) MS using single ion monitoring (SIM) in negative mode. Linear calibration graphs were achieved in the dynamic range of 10-1000 ng/ml urine. The inter- and intraassay coefficients of variation (C.V.%) for the analysis of the four compounds in quality control urine samples were between 7.8 and 10.9, n=17 (reproducibility), and the repeatability of the assay was between 2.5 and 5.0% (n=12). Analyses of urine samples from a human dietary intervention study with intake of 600 g of fruits and vegetables were demonstrated. To our knowledge, this is the first method described that allows simultaneous determination of both hydroxycinnamates and catechins in biological samples. PMID- 12650760 TI - Determination of fifteen nucleotides in cultured human mononuclear blood and umbilical vein endothelial cells by solvent generated ion-pair chromatography. AB - The paper describes the development of a method for the determination of 15 nucleotides in cultured mononuclear blood and umbilical vein endothelial cell lysates by solvent generated ion-pair chromatography. The phase system is generated via a mobile phase of 100 mM phosphoric acid adjusted to pH 6.2 with triethylamine. Nucleotides are eluted by applying a linear magnesium ion gradient. The method is robust, highly reproducible and easily adaptable to other cell lysates and allows the separation and quantitation of the nucleotides with detection limits in the range from 17 (ADP) to 126 (CDP) pmol in 20-microl aliquots. PMID- 12650761 TI - Determination of protease inhibitors using liquid chromatography-tandem mass spectrometry. AB - A method for the analysis of six protease inhibitors and one metabolite has been developed and validated. Amprenavir, ritonavir, saquinavir, lopinavir, indinavir, nelfinavir, and an active metabolite of nelfinavir (M8) are quantitated using reversed-phase liquid chromatography coupled to tandem mass spectrometry, equipped with an electrospray ionization source (ESI-LC-MS-MS). The validation data presented here shows that the method allows the rugged analysis of these species from one aliquot. The evolution of complex drug interactions assessments and the clinical use of therapeutic drug monitoring for these antiretrovirals will be a potential immediate application of this method. PMID- 12650762 TI - Liquid chromatography-electrospray mass spectrometry study of cysteine-10 S glutathiolation in recombinant glutathione S-transferase of Ochrobactrum anthropi. AB - Glutathione S-transferase of Ochrobactrum anthropi (OaGST), a bacterium isolated from soils contaminated by xenobiotic pollutants, was recently purified, cloned and characterised in our laboratories. The recombinant OaGST (rOaGST), highly expressed in Escherichia coli, when purified by glutathione-affinity chromatography and then analysed by electrospray ionisation mass spectrometry (ESI-MS), has evidenced a disulphide bond with glutathione (S-glutathiolation), which was removable by reduction with 2-mercaptoethanol. Enzymatic digestion of rOaGST with endoproteinase Glu-C, followed by liquid chromatography (LC)-ESI-MS analyses of the peptide mixtures under both reducing and not reducing conditions, have shown that glutathione was covalently bound to the Cys10 residue of rOaGST. Furthermore, LC-ESI-MS analyses of overexpressed rOaGST in Escherichia coli crude extracts, with and without incubation with glutathione, have not shown any S glutathiolation of the recombinant enzyme. PMID- 12650763 TI - Simple liquid-chromatographic method for Nile Red quantification in cell culture in spite of photobleaching. AB - Nile Red fluorescent marker is widely-used for different purposes, such as staining cell structures and for the visualization and localization of colloidal drug carriers. However, when fluorescence-dependent imaging or quantification is performed, the risk of inexact results is increased due to photobleaching. The proposed, simple quantification method of using an HPLC-UV-Vis system allows the determination of Nile Red even in photobleached samples. The intra- and inter assay accuracies for all analytes were found to be within 94.9 and 100.8%, respectively, of target values. When samples underwent photobleaching by laser, UV-Vis detection varied at around 99+/-5%, whereas fluorescence decreased down to 86%. Such results show this method to be interesting for approaches where quantification should be performed after analysis such as fluorescent imaging. PMID- 12650764 TI - Determination of spinosin in rat plasma by reversed-phase high-performance chromatography after oral administration of Suanzaoren decoction. AB - A sensitive, simple, and accurate method for determination of spinosin in rat plasma with sulfamethoxazole (SMZ) as internal standard was developed using RP HPLC with UV detection. Sample preparations were carried out by protein precipitation with acetonitrile, followed by the evaporation of the acetonitrile to dryness. The resultant residue was then reconstituted in mobile phase and injected onto a Hypersil C(18) (200 x 4.6 mm I.D., 5 microm) analytical column. The mobile phase consisted of acetonitrile-water (15:85, v/v) with 1% glacial acetic acid. The assay was shown to be linear over the range of 18.07-903.5 ng/ml (R(2)=0.995). Mean recovery was determined as 93.6%. Within- and between-day precisions were 1 month and >2 months after therapy. RESULTS: One hundred thirty eight patients received OBMT and 137 OAC. Modified intent-to-treat eradication rates were 87.7% for OBMT and 83.2% for OAC (95% CI = -3.9%-12.8%; p = 0.29). OBMT eradicated 91.7% metronidazole-sensitive and 80.4% metronidazole-resistant strains (p = 0.06). OAC eradicated 92.1% clarithromycin sensitive and 21.4% clarithromycin-resistant strains (p < 0.001). Adverse events occurred in 58.5% of OBMT patients and 59.0% of OAC patients. CONCLUSIONS: OBMT regimen using the single-triple capsule is as efficacious and well-tolerated as the widely used OAC regimen for H. pylori eradication. This OBMT therapy largely overcomes H. pylori metronidazole resistance, present in 40% of patients in this study. PMID- 12650789 TI - Pancreatic ascites: study of therapeutic options by analysis of case reports and case series between the years 1975 and 2000. AB - OBJECTIVE: Because of the low incidence of pancreatic ascites, only case reports and case series have been published, and no randomized controlled trials have been performed to find out which is the best therapeutic approach. The aim of this study was to evaluate the utility of the different treatments for pancreatic ascites by a thorough review of all case reports described in the literature in which an independent analysis of the efficacy of the treatment administered to each patient is possible. METHODS: We conducted an analysis of all case reports and case series of pancreatic ascites published between 1975 and 2000 in which clinical data of every patient could be identified individually. A total of 139 cases were studied. Clinical characteristics, treatments administered, and response to therapy of every patient were registered. Conservative therapy included drainage of ascitic fluid, total parenteral nutrition and diet, and somatostatin analogues. Interventional therapy was either endoscopic or surgical. RESULTS: After multivariate analysis, the only treatments related to success were surgery (adjusted OR = 8.2, 95% CI = 3.0-22.9) and transpapillary stent (adjusted OR = 7.3, 95% CI = 0.8-62.9). No significant relationship was found between failure or death and the use of other treatments, age, sex, year of publication, underlying disease, site of leakage, or serum amylase levels. The apparent lack of effect of somatostatin analogues could be attributed to the small number of cases and the heterogeneity of the dosages. CONCLUSIONS: Conservative therapy is not advisable for pancreatic ascites because of the high proportion of failures. Interventional therapy with surgery or transpapillary stent has a positive effect in the clinical outcome. PMID- 12650790 TI - Patient preferences for CT colonography, conventional colonoscopy, and bowel preparation. AB - OBJECTIVES: The aim of this study was to determine patient pre-examination expectations and postexamination appraisals for CT colonography, conventional colonoscopy and bowel preparation. METHODS: Prospective evaluation of 120 patients at defined risk for colorectal neoplasia was performed with CT colonography followed by colonoscopy on the same day. Subjects were stratified by age and sex (67 women and 53 men) and were randomized to receive either manual air (n = 61) or CO(2) (n = 59) insufflation during CT colonography. Patients' expectations were assessed just before the two examinations, and appraisals were assessed 2 to 3 days afterward regarding pain/discomfort, embarrassment, difficulty, overall assessment, preference for future testing, and bowel preparation. RESULTS: No significant differences were found in appraisals of manual air versus CO(2) insufflation techniques. For both CT colonography and colonoscopy, patients' appraisals after the procedure were significantly more positive than prior expectations. Patients expressed more favorable appraisals of colonoscopy for pain (p < 0.001) and embarrassment (p < 0.001), with most responses being "none" to "a little" for both examinations. Overall appraisals of the tests were favorable and similar between CT and colonoscopy: patients mainly expressed "not unpleasant" to "a little unpleasant" (95%, 114/120 for both examinations). Overall, appraisal of the bowel preparation was the most negative. Preferences for future testing were more favorable toward CT: of the patients, 58% (69/120) preferred CT, 14% (17/120) preferred colonoscopy, and 28% (34/120) had no preference. CONCLUSIONS: Overall appraisals were similar and positive for both CT colonography and colonoscopy, with less favorable appraisals of the bowel preparation. Most patients stated that they would prefer CT for future evaluation. PMID- 12650791 TI - Endoscopic sphincterotomy in the management of bile duct stones in children. AB - OBJECTIVE: Endoscopic sphincterotomy (ES) is a widely accepted method of extracting bile duct stones (BDS) in young as well as in elderly patients. The present study was undertaken to assess the safety and efficacy of ES for the treatment of BDS in children, seven of whom were critically sick because of suppurative cholangitis or pancreatitis. METHOD: Over a period of 33 months, ES was performed in 16 consecutive children aged 7-16 yr with BDS. Nine patients had gallbladder in situ, and seven had previously undergone cholecystectomy. The coexisting abnormalities were gallstones and hepatic duct stones in one patient each and dead fragmented roundworms in 11 patients. Seven (five with an intact gallbladder and two cholecystectomized) patients presented with severe complications of BDS such as severe cholangitis in six and acute severe pancreatitis in one. RESULTS: ES was technically successful in all patients, and complete stone extraction was achieved in 15 (93.8%) patients. Complications were minor bleeding in one (6.3%) patient without mortality. One patient with coexisting gallstones underwent cholecystectomy at a later date. During a mean follow-up period of 4-32 months, one patient developed recurrent biliary symptoms because of biliary ascariasis. CONCLUSIONS: We conclude ES is a safe and an effective method of treating BDS in children with previous cholecystectomy, and in those presenting with severe complications of BDS, such as pyogenic cholangitis or acute pancreatitis regardless of the presence of gallbladder. PMID- 12650792 TI - Gluten sensitivity in a subset of children with insulin dependent diabetes mellitus. AB - OBJECTIVES: The association between celiac disease and insulin dependent diabetes mellitus (IDDM) is well established. Rectal gluten challenge has been used in patients with celiac disease and in first degree relatives as a tool to assess the mucosal immune response to gluten. The aim of this study was to assess the mucosal immune response to gluten in IDDM children by rectal gluten challenge. METHODS: Rectal biopsy specimens were obtained from 19 children with IDDM before and 6 h after rectal challenge with 2 g of a peptic tryptic digest of gliadin. A total of 16 treated celiac patients and 10 control subjects were also investigated. Epithelium and lamina propria CD3(+) and gamma delta(+) lymphocytes were counted with reference to a standard reference area of muscularis mucosae (10(4) microm(2)). RESULTS: After a local instillation of gliadin, a significant (>mean + 1 SD) percentage increment of lamina propria and epithelium CD3(+) and of lamina propria and epithelium gamma delta(+) lymphocytes was observed in five IDDM children, as compared to 11 and 13 celiac patients and one and two controls, respectively. A discriminant analysis allowed correct classification of 100% of patients with celiac disease and controls. The same analysis classified four of 19 IDDM children in the group of celiac patients. The positivity was associated with normal serology (antigliadin antibody, antiendomysial antibody, and antitissue transglutaminase antibodies) and a morphologically normal jejunal mucosa. All four patients had HLA-DQ alleles associated with celiac disease. CONCLUSIONS: Approximately 20% of IDDM children react to rectal instillation of gliadin. Long term follow-up is necessary to establish whether these subjects are at increased risk for developing celiac disease. PMID- 12650793 TI - Post-traveler's diarrhea irritable bowel syndrome: a prospective study. AB - OBJECTIVE: Anecdotal evidence suggests that irritable bowel syndrome (IBS) can develop after an episode of traveler's diarrhea (TD). This observation supports a contemporary paradigm proposed for the etiology of IBS and may have important implications for public health strategies aimed at preventing TD. This study aimed to determine the incidence of IBS in people experiencing TD. METHODS: A total of 109 healthy adults traveling outside of Canada or the United States were identified and enrolled in a prospective, cohort field study. GI symptoms before and after travel were assessed using the Rome I criteria. Travel diaries and questionnaires were used to assess for TD. RESULTS: The incidence of TD in the study cohort was 44%. Among those experiencing TD, the incidence of IBS was 4.2%. In those not experiencing TD, the incidence of IBS post-travel was 1.6% (relative risk = 2.5, 95% CI = 0.2-27.2, p = 0.41, ns). There was no significant difference in the incidence of IBS between the exposed and nonexposed groups. CONCLUSIONS: This study does not support a large association between TD and an increased risk of developing IBS. A small increase in relative risk may have been undetected because of the size of the study. PMID- 12650794 TI - Irritable bowel syndrome, health care use, and costs: a U.S. managed care perspective. AB - OBJECTIVE: We performed an evaluation of patient symptoms, health care use, and costs to define the burden of illness of irritable bowel syndrome (IBS) and the relation to the severity of abdominal pain/discomfort in a large health maintenance organization. METHODS: All 6500 adult health maintenance organization members who had undergone flexible sigmoidoscopy in the year 2000 were mailed a questionnaire that elicited Rome I symptom criteria and severity ratings for abdominal pain/discomfort. Multiple health care use measures were obtained from various administrative databases. IBS patients were compared with a control group of non-IBS subjects, and analyses were adjusted for age and sex. RESULTS: We received 2613 (40.2%) responses. Compared with non-IBS subjects over 2 yr, IBS patients had more outpatient visits (medical, surgery, and emergency, p < 0.05), were hospitalized more often (p < 0.05), and had more total outpatient prescriptions (p < 0.05) and IBS-related prescriptions (p < 0.05). Over 1 yr, total costs were 51% higher in IBS patients, who also had higher costs for outpatient visits, drugs, and radiology and laboratory tests (p < 0.05). Total costs were increased by 35%, 52%, and 59% in IBS patients with mild, moderate, and severe symptoms of abdominal pain/discomfort compared with non-IBS subjects (p < 0.05). CONCLUSIONS: Using Rome I symptom criteria, we found that IBS is associated with a broad pattern of increased health care use and costs. The severity of abdominal pain/discomfort is a significant predictor of health care use and costs for patients with IBS compared with non-IBS subjects. PMID- 12650795 TI - Immunomodulators and "on demand" therapy with infliximab in Crohn's disease: clinical experience with 400 infusions. AB - OBJECTIVES: Infliximab has been proven effective for treatment of active Crohn's and fistulizing Crohn's disease. We reviewed our experience with infliximab in patients with Crohn's disease to determine if its combination with immunomodulators leads to better response and longer periods of disease quiescence. METHODS: We performed a retrospective chart review of 122 patients with Crohn's disease who received infliximab infusions. Data were collected on patient demographics, clinical response to infliximab, fistula response, prednisone dose, infusion reactions/side effects, concomitant immunomodulator therapy, and time intervals between infliximab infusions. RESULTS: Of 122 patients receiving infliximab infusions, 117 completed more than 2 wk of follow up (400 infusions), and five patients had no follow-up. Co-therapies included azathioprine (AZA) in 47 (40.2%) patients, 6-mercaptopurine (6-MP) in 11 (9.4%), methotrexate (MTX) in 23 (19.7%), prednisone in 64 (54.7%), mesalamine in 51 (43.6%), and antibiotics in 16 (13.7%). Mean follow-up was 52 wk (14-864 days). Overall response rate to infliximab was similar between patients receiving immunomodulators (AZA/6-MP 87.9%, MTX 82.6%) and patients receiving infliximab alone (75%), although there was a trend toward higher response with AZA/6-MP (p = 0.10). More frequent drug reactions/side effects occurred in the infliximab alone group (22.2%) compared with patients receiving MTX (13.0%) and AZA/6-MP (13.8%), but this was not statistically significant. Prednisone dosage was reduced from a mean of 19.5 mg to 7.5 mg per day overall (p < 0.05). Fistula response and dosing intervals were not affected by concomitant immunosuppression. CONCLUSIONS: Concomitant use of immunomodulators with infliximab in patients with Crohn's disease did not improve patient response to several parameters measured, including clinical response rate, dose reduction of prednisone, fistula response, and mean intervals between infliximab infusions. PMID- 12650796 TI - Card15 and Crohn's disease: healthy homozygous carriers of the 3020insC frameshift mutation. AB - OBJECTIVES: Single nucleotide variations in the CARD15 gene have recently been shown to be associated with Crohn's disease (CD). Of special interest is a cytosine insertion at position 3020 of exon 11 (3020insC), which leads to a stop codon, truncation of the CARD15 protein, and an altered function of CARD15. The aim of the study was to evaluate this frameshift mutation in Dutch, multiple affected families with inflammatory bowel disease (IBD). METHODS: Ninety-three Caucasian, multiple-affected families with IBD were recruited by interviewing patients attending our department. Sixty-one probands had CD, and 32 probands ulcerative colitis (UC). The diagnosis of probands and affected family members was verified according to standard criteria. In addition, 81 healthy, unrelated controls were included. Genomic DNA was isolated from venous blood of all participants to determine the CARD15 3020insC mutation by using an allele specific polymerase chain reaction, followed by agarose gel electrophoresis and DNA sequencing. RESULTS: Association with CARD15 3020insC was statistically significant for CD, but not for UC. In one of the multiple-affected families, middle-aged and elderly homozygous carriers were identified without CD. CONCLUSIONS: Although CARD15 3020insC appears to be etiologically important in CD, homozygous carriage does not always lead to IBD. PMID- 12650797 TI - Role of interstitial cells of Cajal in motility disorders of the bowel. AB - OBJECTIVE: Idiopathic intestinal pseudo-obstruction is characterized by the failure of the intestinal tract to propel its contents appropriately. This leads to signs and symptoms of bowel obstruction and, in the absence of an associated systemic disorder or the administration of drugs known to result in bowel dysmotility, is termed chronic idiopathic intestinal pseudo-obstruction (CIIP). Histopathologically, patients with CIIP can be characterized as having either myopathic or neuropathic forms, but the large majority of patients do not show any specific histological changes. Interstitial cells of Cajal (ICC) have been shown to be the pacemaker cells of the bowel and have been implicated in the pathogenesis of CIIP. The aim of this study was to compare the number and distribution patterns of c-kit+ ICC in CIIP in patients with mechanical bowel obstruction, other bowel motility disorders, and normal controls. METHODS: Six patients with CIIP, six age-matched normal controls, nine patients with mechanical bowel obstruction, and 18 patients with other motility disorders (non CIIP), including 10 with secondary intestinal pseudo-obstruction, were studied. Toluidine blue, Masson's trichrome, and S-100 immunostaining were performed in all subjects. The ICC were identified by an indirect immunoperoxidase method using a polyclonal c-kit antibody. RESULTS: All six patients with CIIP showed total absence of c-kit+ ICC. A subject with neonatal meconium ileus in the non CIIP group showed patchy areas devoid of c-kit+ ICC amid normal areas. The c-kit+ ICC had a normal number and distribution pattern in all patients with mechanical obstruction and in the remaining 17 non-CIIP subjects. CONCLUSIONS: It seems that CIIP is characterized by a total loss of c-kit+ ICC. ICC may play an important role in the etiopathogenesis of CIIP and transient neonatal meconium syndrome, and staining for c-kit receptor may be very useful in the evaluation of motility disorders of the bowel. PMID- 12650798 TI - Association between migraine and Celiac disease: results from a preliminary case control and therapeutic study. AB - OBJECTIVES: Subclinical celiac disease (CD) has been associated with various neurological disorders, the most common being neuropathy and cerebellar ataxia. The aims of the present study were to assess the following: 1) the prevalence of CD in patients affected by migraine; 2) whether there are regional cerebral blood flow abnormalities in migraine patients with CD compared to migraine patients without CD; and 3) the effects of a gluten free diet in migraine patients with CD. METHODS: A total of 90 patients affected by idiopathic migraine were enrolled, and 236 blood donors were used as controls. Serum IgG antitransglutaminase (TgA) and IgA antiendomysial (EmA) were measured. In positive cases, diagnosis was confirmed endoscopically. A gluten free diet was started in the patients diagnosed with CD, who were followed for 6 months. A single photon emission CT brain study was performed before and after a gluten free diet. RESULTS: Four of 90 (4.4%; 95% CI = 1.2-11.0) migraine patients were found to have CD compared with 0.4% (95% CI = 0.01-2.3) blood donor controls (p < 0.05). During the 6 months of gluten free diet, one of the four patients had no migraine attacks, and the remaining three patients experienced an improvement in frequency, duration, and intensity of migraine. Single photon emission CT studies showed a regional baseline reduction in brain tracer uptake in all four patients. Such reduction in uptake completely resolved at follow-up. CONCLUSIONS: Our results suggest that a significant proportion of patients with migraine may have CD, and that a gluten free diet may lead to a improvement in the migraine in these patients. PMID- 12650799 TI - Health-state utilities and quality of life in hepatitis C patients. AB - OBJECTIVE: Health-state utilities are global measurements of quality of life on a scale from 0 (death) to 1 (full health). Utilities are used to evaluate health outcomes and are the preferred outcome measure for policy models that determine the cost-effectiveness of treatments. Currently, utilities for hepatitis C virus (HCV)-infected patients have been estimated using expert judgments. The purpose of this study was to elicit HCV utilities directly from patients. METHODS: We assessed the utilities of 193 outpatients at various stages of chronic HCV progression by using a visual analog scale, the standard gamble technique, the Health Utilities Index Mark 3 survey, and the EuroQol Index survey. We also incorporated the nonutility-based Short Form-36v2 survey, which provides a detailed profile of health status. RESULTS: The mean standard gamble utilities were: 0.78 for patients without a recent liver biopsy and no signs of cirrhosis; 0.79 for mild to moderate chronic HCV infection; 0.80 for compensated cirrhosis; 0.60 for decompensated cirrhosis; 0.72 for hepatocellular carcinoma; 0.73 for transplant; and 0.86 for sustained virological responders to interferon +/- ribavirin treatment. The Health Utilities Index Mark 3 survey and the EuroQol Index survey utilities were lower than Canadian population norms (p < 0.001). Patient-elicited utilities were lower than previous expert estimates for mild/moderate chronic infection and sustained virological responders, but higher for decompensated cirrhosis and hepatocellular carcinoma. The Short Form-36v2 survey scores revealed several significant health impairments (p < 0.005) when compared with U.S. population norms. CONCLUSIONS: These findings 1) suggest that quality of life (QOL) differences across the HCV clinical spectrum are smaller than previously believed; 2) support other evidence suggesting that QOL is significantly diminished in HCV patients; and 3) provide utility values derived directly from HCV patients. PMID- 12650800 TI - Identification and management of hepatitis C patients in primary care clinics. AB - OBJECTIVE: Previous survey-based research suggested that hepatitis C patients receive suboptimal care in primary care settings. The aim of our study was to define the actual level of care hepatitis C patients receive in primary care clinics. METHODS: Medical records of 229 hepatitis C antibody-positive (group 1), 229 hepatitis C antibody-negative (group 2), and 229 patients not tested for hepatitis C antibody (group 3) were reviewed to assess the indications for hepatitis C testing and the subsequent management and referral of hepatitis C antibody-positive patients diagnosed in primary care clinics. In addition, the compliance of primary care physicians with hepatitis C screening and testing guidelines was assessed. RESULTS: Only 16% of group 1 and 10% of group 2 patients were tested for hepatitis C based on physician-identified risk factors. Only 1% of group 3 patients had documented discussion of hepatitis C risk factors during their initial visit with a primary care physician. The majority of hepatitis C antibody-positive patients was appropriately evaluated in primary care clinics, and most (77%) hepatitis C RNA-positive patients with elevated liver enzymes were referred for subspecialty care. Of the 59 patients who underwent liver biopsy, 40% had bridging fibrosis or cirrhosis. CONCLUSIONS: Hepatitis C testing is rarely initiated in primary care clinics based on physician-identified risk factors. Interventions should be developed to optimize early diagnosis of hepatitis C as significant liver disease may be present despite the absence of symptoms. PMID- 12650801 TI - Nonimmune complexed HCV RNA titer in serum as a predictor of interferon response in patients with chronic hepatitis C. AB - OBJECTIVE: Hepatitis C virus (HCV) has been reported to exist in the circulation of patients in various forms such as free virions, immune complexes, and nucleocapsids. To clarify the clinical significance of serum HCV titers according to the forms of virus particles, we evaluated the immune complexed (IC) and nonimmune complexed (NIC) HCV RNA titers in 77 chronic hepatitis patients treated with interferon (IFN). METHODS: IC and NIC forms in pretreatment serum were separated by immunoprecipitation using antihuman immunoglobulin antibody, and quantified by reverse transcription polymerase chain reaction. RESULTS: Serum titers of NIC HCV RNA were correlated with those of whole serum HCV RNA (r = 0.96, p < 0.01) and IC HCV RNA (r = 0.98, p < 0.01), but they were not with the aminotransferase levels, gamma-globulin concentration, and grading or staging of liver histology. Nonresponders to IFN had significantly high NIC HCV RNA titers compared with sustained responders (10(4.93 +/- 0.81) copies/ml vs 10(4.06 +/- 0.69) copies/ml, p < 0.01). It is noteworthy that the relative amount of NIC HCV RNA to whole serum HCV RNA was also significantly higher in nonresponders than in sustained responders (0.66 +/- 0.10 vs 0.50 +/- 0.11, p < 0.0001). Multivariate analysis showed that low NIC HCV RNA titer (p < 0.01) and genotype 2 (p = 0.02) were independent variables contributing to sustained response to IFN, but the whole serum HCV RNA titer was not. CONCLUSIONS: Pretreatment NIC HCV RNA titer is a more reliable predictive marker than genotype or whole serum HCV RNA of a sustained response to IFN monotherapy. This finding suggests that humoral immunity may affect the response to IFN. PMID- 12650802 TI - Improved patient survival after acute variceal bleeding: a multicenter, cohort study. AB - OBJECTIVE: Existing literature indicates that the mortality rate with each variceal bleeding episode is 30-50%. Over the past 2 decades, there have been significant developments in the management of variceal bleeding. The effect of these developments on the natural history of variceal bleeding is unclear. Therefore, a retrospective, multicenter study was conducted to define the outcomes of variceal bleeding and to describe the patterns of current practice in the management of variceal bleeding. METHODS: All patients with documented variceal bleeding hospitalized at four large county hospitals from January 1, 1997, to June 30, 2000, were included. Study outcomes were in-hospital, 6-wk, and overall mortality, rate of rebleeding, transfusion requirement, and length of stay. After discharge, patients were followed until death or study closure date, on June 30, 2000. RESULTS: A total of 231 subjects were included, and their in hospital, 6-wk, and overall mortality rates were 14.2%, 17.5%, and 33.5%, respectively. The frequency of rebleeding during follow-up was 29%. Median length of total hospital stay was 8 days (0-34 days). Median number of packed red cell units transfused was 4 U (0-60 U). Upper endoscopy was performed in 95% of patients within 24 h, and endoscopic therapy was done in all but eight patients (ligation 64%, sclerotherapy 33%). Octreotide was administered in 74% of the patients. Portasystemic shunts were performed in 7.5% of the patients for controlling acute variceal bleeding. CONCLUSIONS: The mortality rate after variceal bleeding in this study was substantially lower than previously reported. This suggests that advances made in the management of variceal bleeding have improved outcomes after variceal bleeding. PMID- 12650803 TI - Fibrosis in chronic hepatitis C acquired in infancy: is it only a matter of time? AB - OBJECTIVE: The natural history of chronic hepatitis C acquired in infancy is not well understood. The progression of fibrosis was analyzed in untreated children with chronic hepatitis C virus infection and no other hepatotoxic cofactors. METHODS: A total of 112 pediatric patients (13 with paired liver biopsies) were considered. Fibrosis was assessed by METAVIR score (i.e., stage F1 to F4). The ratio between the stage of fibrosis (METAVIR units) and the presumed duration of infection represented the "estimated" rate of fibrosis progression per year. In patients with paired biopsies, the "observed" rate of fibrosis progression was defined as the difference between the stage of fibrosis in the two biopsies divided by the time interval between them. RESULTS: Both age of patients at biopsy and duration of infection correlated with stage of fibrosis (p < 0.002 and p < 0.0005, respectively). Stage of fibrosis differed significantly between patients with infection lasting less or more than 10 yr (p < 0.0006). Sex, hepatitis C virus genotype, and route of infection did not correlate with stage of fibrosis. Among the 13 patients with paired biopsies, stage of fibrosis increased in seven and did not change in six; the median rate of estimated fibrosis progression per year was 0.142. The difference between estimated and observed fibrosis progression rates was significant (coefficient of determination, r(2) = 0.031), which demonstrated that the prediction of the fibrosis progression was unreliable in 97% of patients. CONCLUSIONS: Chronic hepatitis C acquired in childhood is a progressive, slow-moving, fibrotic disease. Fibrosis progression inferred on the basis of linear mathematical models should be critically evaluated in the clinical practice. PMID- 12650804 TI - Multiple primary cancer, including transitional cell carcinoma of the upper uroepithelial tract in a multigeneration HNPCC family: molecular genetic, diagnostic, and management implications. AB - OBJECTIVE: We report a multigeneration family where colorectal cancer and cancer of multiple diverse anatomic sites, inclusive of transitional cell carcinoma of the upper uroepithelial tract, were manifested in several relatives. METHODS: A specific pattern of cancer of the colorectum, endometrium, ovary, small bowel, and transitional cell carcinoma, with a vertical distribution of this cancer phenotype through multiple generations, was consonant with a diagnosis of hereditary nonpolyposis colorectal cancer. RESULTS: Germline mutation testing identified the MSH2 mutation, which segregated with the cancer phenotype. This family study clearly demonstrates the value of genetic testing in the management and treatment decision process. CONCLUSIONS: We document, perhaps for the first time, how molecular genetic testing in hereditary nonpolyposis colorectal cancer can aid in the identification of a potential renal transplant donor for a relative with the MSH2 mutation who is experiencing renal insufficiency secondary to transitional cell carcinoma. PMID- 12650805 TI - Overexpression of cyclooxygenase 2 in hamartomatous polyps of Peutz-Jeghers syndrome. AB - OBJECTIVE: Peutz-Jeghers syndrome (PJS) is an autosomal-dominant hamartomatous polyposis syndrome. Affected individuals are at risk for intestinal and extraintestinal malignancies. Prostaglandins and polyamines are small molecules believed to be important in tumor formation and growth. Cyclooxygenase (COX) and ornithine decarboxylase (ODC) are key enzymes in the prostaglandin and polyamine biosynthetic pathways, respectively. The aim of this study was to measure and compare COX-1 and COX-2 expression in normal and hamartomatous tissue of PJS patients. METHODS: We measured COX-1 and COX-2 protein expression in normal and hamartomatous GI tissues from affected PJS individuals and compared it with that in normal controls. COX-2 RNA in these tissues was also measured and compared by reverse transcription polymerase chain reaction (PCR). In addition, COX-2 expression was detected in tissue slides by immunostaining. ODC activity was measured between normal and hamartomatous tissues of PJS compared with control tissues. RESULTS: COX-1 expression was similar in normal and control GI tissues. In contrast, COX-2 overexpression was noted in hamartomatous polyp tissue from PJS patients compared with normal control and PJS tissue. COX-2 expression by reverse transcription PCR was 10-fold greater in a hamartoma compared with other tissues. COX-2 expression was noted in the epithelial cells of hamartomatous polyps, and also coursing throughout the stromal tissue of the lamina propria, including muscle cells. ODC activity was similar in the tissues studied. CONCLUSIONS: Selective COX-2 overexpression was noted in hamartomatous polyp tissue from PJS individuals. The results of the study provide an avenue for possible effective chemoprevention of polyp formation and growth in PJS. PMID- 12650807 TI - Chemotherapy for MALTomas--are we back to where we started? PMID- 12650808 TI - Colonoscopy: go small for success? PMID- 12650806 TI - Screening for hepatocellular carcinoma in patients with hepatitis C cirrhosis: a cost-utility analysis. AB - OBJECTIVES: Screening for hepatocellular carcinoma (HCC) is advocated in cirrhotic patients to optimize early detection and treatment. However, the cost effectiveness is not well defined. Our objective was to perform a cost-utility analysis from a third-party payer's perspective of no screening, alpha fetoprotein (AFP) concentration measurement alone, abdominal ultrasound (US) and AFP, abdominal three-phase CT and AFP, and abdominal magnetic resonance imaging (MRI) and AFP. METHODS: A Markov model was constructed simulating the natural history of hepatitis C-related cirrhosis in a cohort of patients age 50 yr over a time horizon of their remaining life expectancy. Transition probabilities were obtained from published data and U.S. vital statistics. Costs represented Medicare reimbursement data. Costs and health effects were discounted at a 3% annual rate. RESULTS: Screening with ultrasonography and AFP concentration measurement was associated with an incremental cost-utility ratio of 26,689 US dollars per quality-adjusted life year, whereas screening with abdominal three phase CT and AFP concentration measurement was associated with an incremental cost-utility ratio of 25,232 US dollars per quality-adjusted life year compared with no screening. Compared with three-phase CT and AFP, magnetic resonance and AFP imaging costs 118,000 US dollars per quality-adjusted life year. Sensitivity analysis demonstrated that the results are most sensitive to the annual incidence of HCC, proportion of tumors amenable to treatment, and to transplant candidacy, whereas the choice of screening strategy is most sensitive to the test characteristics and cost. CONCLUSIONS: Screening for HCC with CT is a cost effective strategy in transplant-eligible patients with cirrhosis secondary to chronic hepatitis C viral (HCV) infection, comparable with other commonly accepted screening interventions such as mammography and colonoscopy. PMID- 12650809 TI - NSAIDS: can we stomach the risk? PMID- 12650810 TI - Diagnosis and follow-up of patients with achalasia. PMID- 12650811 TI - Cardiac achalasia. PMID- 12650813 TI - Treatment of ulcerative colitis with balsalazide: response to Drs Johnson, Green, and Pruitt. PMID- 12650814 TI - Gallbladder perforation associated with metastatic invasion from esophageal carcinoma. PMID- 12650815 TI - Significance of acid exposure in Barrett's esophagus. PMID- 12650816 TI - The helpful colon. PMID- 12650817 TI - Familial Mediterranean fever mimicking Fitz-Hugh-Curtis syndrome. PMID- 12650818 TI - Right testicular varicocele: an unusual presentation of cecal adenocarcinoma. PMID- 12650819 TI - Successful treatment of cervical esophageal perforation in Behcet's disease with drainage operation and infliximab. PMID- 12650820 TI - A case of montelukast-induced hepatotoxicity. PMID- 12650821 TI - Pegylated interferon-2b-associated autoimmune thrombocytopenia in a patient with chronic hepatitis C. PMID- 12650822 TI - Celiac disease and IBS-type symptoms: the relationship exists in both directions. PMID- 12650823 TI - Repeated neostigmine dosage as palliative treatment for chronic colonic pseudo obstruction in a patient with autonomic paraneoplastic neuropathy. PMID- 12650824 TI - Infliximab for ulcerative colitis. PMID- 12650825 TI - Rupture of an aneurysm of the right external iliac artery as a complication of colonoscopy. PMID- 12650826 TI - Interaction of human and rat organic anion transporter 2 with various cephalosporin antibiotics. AB - Cephalosporin antibiotics are thought to be excreted into the urine via organic anion transporters (OATs) and OAT can mediate nephrotoxicity by cephalosporins, particularly by cephaloridine. The purpose of this study was to elucidate the interaction of human-OAT2 and rat-OAT2 with cephalosporin antibiotics using proximal tubule cells stably expressing human-OAT2 and rat-OAT2. Human-OAT2 is localized to the basolateral side of the proximal tubule, whereas rat-OAT2 is localized to the apical side of the proximal tubule. Cephalosporins tested were cephalothin, cefoperazone, cefazolin, ceftriaxone, cephaloridine, cefotaxime, cefadroxil and cefamandole. These cephalosporins dose-dependently inhibited organic anion uptake mediated by human-OAT2 and rat-OAT2. There was no species difference observed for the effects of OAT2 with cephalosporins between human and rat transporters. Kinetic analysis revealed that the inhibitory effects for human OAT2 were competitive. Cephaloridine significantly decreased the viability of cells stably expressing human-OAT2, human-OAT1, human-OAT3 and human-OAT4. The decreased viability of cells stably expressing human-OAT1, human-OAT3 and human OAT4 but not human-OAT2 was reversed by probenecid. In conclusion, human-OAT2 interacts with cephalosporins, and thus, human-OAT2 may mediate the uptake of cephalosporins on the basolateral side of the proximal tubule. The interaction of human-OAT2 with cephalosporins was the weakest among the basolateral human-OATs tested. In addition, it is suggested that human-OATs mediate cephaloridine induced nephrotoxicity. PMID- 12650827 TI - Palytoxin-induced increase in cytosolic-free Ca(2+) in mouse spleen cells. AB - The effect of palytoxin (C(129)H(223)N(3)O(54)) on Ca(2+) homeostasis in immune cells has not been studied. Therefore, we investigated the effect of palytoxin on the cytosolic-free Ca(2+) concentration ([Ca(2+)](i)) in mouse spleen cells using a fluorescence Ca(2+) indicator, fura-2. Palytoxin (0.1-100 nM) increased [Ca(2+)](i) in a concentration-dependent manner. The palytoxin-induced increase in [Ca(2+)](i) was abolished by the omission of extracellular Ca(2+) or 1-[beta [3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H-imidazole hydrochloride (SKF 96365, 100 microM), and was greatly inhibited by Ni(2+) (2 mM). Ouabain (0.5-1 mM) partially inhibited the palytoxin-induced response. There was no effect of decreased extracellular Na(+) (6.2 mM), tetrodotoxin (1 microM), verapamil (10 microM), nifedipine (10 microM), omega-agatoxin IVA (200 nM), omega-conotoxin GVIA (1 microM), omega-conotoxin MVIIC (500 nM), or La(3+) (100 microM). These results suggest that palytoxin increases [Ca(2+)](i) in mouse spleen cells by stimulating Ca(2+) entry through an SKF-96365-, Ni(2+)-sensitive pathway. PMID- 12650829 TI - Early changes in metabolism of leukemic cell lines upon induction of apoptosis by cytotoxic drugs. AB - We evaluated real-time changes in extracellular acidification rates of human U937 and K562 leukemic cells treated with camptothecin or taxol. U937 cells treated with camptothecin or taxol for 30-60 min showed a continuous, irreversible decrease in extracellular acidification rate that was sensitive to amiloride. In contrast, U937 cells exposed to sodium azide showed an immediate, steep decrease in extracellular acidification rate that was reversible upon azide withdrawal. K562 cells required a >20-fold higher dose of camptothecin to promote similar changes in the extracellular acidification rate, with a corresponding resistance in their susceptibility to camptothecin- or taxol-induced apoptosis. The data show that irreversible commitment to apoptosis is associated with rapid metabolic changes that are reflected by decreased extracellular acidification rate and regulated by the Na(+)/H(+) antiporter. Moreover, detection of extracellular acidification rate changes was not restricted to a particular cell type or apoptosis pathway, making this a potentially useful tool to screen compounds for pro-apoptotic activity. PMID- 12650828 TI - Neuroprotective effects of alpha-tocopherol on oxidative stress in rat striatal cultures. AB - Oxidative stress caused by an increase in free radicals plays an important role in neuronal death. We investigated the effects of alpha-tocopherol on oxidative stress-induced cytotoxicity using primary cultures of rat striatal neurons. alpha Tocopherol at concentrations of 1-10 microM significantly prevented cytotoxicity induced by superoxide radical (O(2(-)) donor, 1,1'-dimethyl-4,4'-bipyridium dichloride (paraquat). In contrast, alpha-tocopherol did not affect the cytotoxicity of hydrogen peroxide (H(2)O(2)), which enhances hydroxyl radical (.OH) formation by metal-catalyzed Fenton reactions. alpha-Tocopherol significantly inhibited the cytotoxicity of nitric oxide (NO) donors, S nitrosocysteine and 3-morpholinosydnonimine (SIN-1). alpha-Tocopherol showed potent protection against cytotoxicity induced by L-buthionine-[S,R]-sulfoximine (BSO), which causes depletion of intracellular glutathione. Moreover, alpha tocopherol afforded a moderate but significant inhibition of cytotoxicity induced by a non-specific protein kinase inhibitor, staurosporine, which is known to induce apoptosis in many types of cells including neurons. These results suggest that alpha-tocopherol protects striatal neurons by the reduction of oxidative stress, presumably by decreasing intracellular O(2)(-) levels, and at least partly by the inhibition of apoptosis. PMID- 12650830 TI - Binding of a new bioactive 31-amino-acid endothelin-1 to an endothelin ET(B) or ET(B)-like receptor in porcine lungs. AB - Endothelin-1-(1-31) is a new bioactive 31-amino-acid-length peptide generated from big endothelin-1 by chymase or other chymotrypsin-type proteases with various pathophysiologic functions. In this study, we have detected the specific and monophasic binding of [125I]endothelin-1-(1-31) in porcine lung membranes. Competition studies of [125I]endothelin-1-(1-31) binding by unlabeled endothelin 1-(1-31), endothelin-1, endothelin-3, and antagonists and agonists of endothelin ET(A) and ET(B) receptors suggest that the binding protein is an endothelin ET(B) or ET(B)-like receptor rather than an endothelin ET(A) receptor in porcine lungs. Kinetic studies showed that the affinity of endothelin-1-(1-31) to its receptor was approximately one order of magnitude lower than that of endothelin-1, and that the specific binding of endothelin-1-(1-31) was about 19% of endothelin-1 binding. The binding of [125I]endothelin-1-(1-31) was extremely slow, slower even than that of endothelin-1, and nearly irreversible. This unique quasi irreversibility may explain the slow-onset and long-lasting biologic effects of this peptide in vivo. PMID- 12650831 TI - Prostaglandin F(2alpha) stimulation of cyclooxygenase-2 promoter activity by the FP(B) prostanoid receptor. AB - We have recently shown that the FP(B) prostanoid receptor activates beta-catenin signaling through the activation of Rho in human embryo kidney (HEK)-293 cells stably expressing the FP(B) receptors. We now report that the FP(B) receptor can stimulate cyclooxygenase-2 promoter activity and may, therefore, regulate the expression of cyclooxygenase-2. This stimulation of cyclooxygenase-2 promoter activity is blocked by pretreatment with an inhibitor of Rho, but not with an inhibitor of protein kinase C (PKC). Potential up regulation of cyclooxygenase-2 expression by the FP(B) receptor would establish a positive feedback loop that would drive beta-catenin signaling and could be involved in cancer. PMID- 12650832 TI - Synergistic interaction between felbamate and lamotrigine against seizures induced by 4-aminopyridine and pentylenetetrazole in mice. AB - We compared the effects of adding a nonprotective dose of felbamate to increasing single doses of lamotrigine with those of monotherapy and vice versa in CD1 mice. Anticonvulsant effects were evaluated against seizures induced by both 14 mg/kg of 4-aminopyridine and 110 mg/kg of pentylenetetrazole, and neurotoxic effects were evaluated by the rotarod test. Changes in anticonvulsants, gamma aminobutyric acid (GABA) and glutamate concentrations in the whole brain were also assessed. Lamotrigine increased the potency ratio of felbamate against 4 aminopyridine (1.80, 95% confidence interval (CI) 1.23-2.65, P<0.05) but not against pentylenetetrazole nor on rotarod, the protective index being increased from 12.0 to 17.1 for 4-aminopyridine, with a reduction in brain felbamate, and with an increase in brain GABA. Felbamate increased the potency ratio of lamotrigine against 4-aminopyridine (4.35, 95% CI 2.05-9.25, P<0.05) but not on rotarod, the protective index being increased from 4.4 to 15.7; there were no changes in brain lamotrigine, and changes in brain GABA and/or glutamate were unrelated to the pharmacodynamic effects. In conclusion, a nonprotective dose of lamotrigine increased the therapeutic index of felbamate and vice versa, and these effects appeared to be pharmacodynamic. PMID- 12650833 TI - Effects of serine/threonine protein phosphatase inhibitors on morphine-induced antinociception in the tail flick test in mice. AB - The aim of this study was to evaluate the effects of serine/threonine protein phosphatase (PP) inhibitors on morphine-induced antinociception in the tail flick test in mice, and on [3H]naloxone binding to the forebrain crude synaptosome fraction. Neither okadaic acid nor cantharidin (1-10000 nM) displaced [3H]naloxone from its specific binding sites, which indicates that they do not interact at the opioid receptor level. The i.c.v. administration of very low doses of okadaic acid (0.001-1 pg/mouse) and cantharidin (0.001-1 ng/mouse), which inhibit PP2A, produced a dose-dependent antagonism of the antinociception induced by morphine (s.c.). However, L-nor-okadaone (0.001 pg/mouse-1 ng/mouse, i.c.v.), an analogue of okadaic acid lacking activity against protein phosphatases, did not affect the antinociceptive effect of morphine. On the other hand, high doses of okadaic acid (10 ng/mouse, i.c.v.) and cantharidin (1 microg/mouse, i.c.v.), which also block PP1, and calyculin-A (0.1 fg/mouse-1 ng/mouse, i.c.v.), which inhibits equally both PP1 and PP2A, did not modify the morphine-induced antinociception. These results suggest that the activation of type 2A serine/threonine protein phosphatases may play a role in the antinociceptive effect of morphine, and that PP1 might counterbalace this activity. PMID- 12650834 TI - GABAergic modulation of hippocampal glutamatergic neurons: an in vivo microdialysis study. AB - We have demonstrated the effects of activation of presynaptic gamma-aminobutyric acid (GABA) receptors on glutamate release using in vivo brain microdialysis. A dialysis probe inserted into the hippocampus CA2 area of freely moving rats was perfused with Ringers solution containing 100 mM potassium chloride (KCl) or 0.05 mM veratridine for 20 min. Extracellular concentrations of amino acids were monitored by measuring their levels in dialysates by high performance liquid chromatography (HPLC) fluorometry. Perfusion with depolarizing agents, such as KCl or veratridine, increased extracellular glutamate levels in the hippocampus. Pretreatment with 1 mM GABA, before perfusion with depolarizing agents, significantly suppressed the depolarizing agent-induced increase in glutamate levels. The GABA(B) receptor agonist baclofen (1 mM) also significantly inhibited the depolarizing agent-induced increase in glutamate levels, whereas the GABA(A) receptor agonist, muscimol, had no affect. Similarly, baclofen (0.5 mM) decreased the KCl (13.5 mM)-induced 45Ca(2+) influx into cortical synaptosomes to 57% of the level induced in the absence of baclofen. On the other hands, GABA did not affect the increases in glycine and taurine level by depolarizing agents. These results suggest that GABA modulates depolarization-evoked glutamate release in the hippocampus by inhibiting Ca(2+) entry into neurons, an effect mediated by presynaptic GABA(B) receptors. PMID- 12650835 TI - Effects of nandrolone on acute morphine responses, tolerance and dependence in mice. AB - Anabolic-androgenic steroid exposure has been proposed to present a risk factor for the misuse of other drugs of abuse. We now examined whether the exposure to the anabolic-androgenic steroid, nandrolone, would affect the acute morphine responses, tolerance and dependence in rodents. For this purpose, mice received nandrolone using pre-exposure (for 14 days before morphine experiments) or co administration (1 h before each morphine injection) procedures. Nandrolone treatments increased the acute hypothermic effects of morphine without modifying its acute antinociceptive and locomotor effects. Nandrolone also attenuated the development of tolerance to morphine antinociception in the hot plate test, but did not affect tolerance to its hypothermic effects, nor the sensitisation to morphine locomotor responses. After nandrolone pre-exposure, we observed an attenuation of morphine-induced place preference and an increase in the somatic manifestations of naloxone-precipitated morphine withdrawal. These results indicate that anabolic-androgenic steroid consumption may induce adaptations in neurobiological systems implicated in the development of morphine dependence. PMID- 12650836 TI - Nicotinic-glutamatergic interactions and attentional performance on an operant visual signal detection task in female rats. AB - Nicotinic systems have been shown to be critically involved in cognitive function including attention. Nicotine has been shown to improve performance on attentional tasks in humans with Alzheimer's disease, schizophrenia and attention deficit hyperactivity disorder. Nicotine has mixed effects on attentional accuracy in unimpaired rats with findings of increased, reduced or unaltered accuracy under different conditions. Nicotine effects on attentional function in rats might be more clearly seen in reversing impaired performance. The current study determined nicotine effects on attentional accuracy reduced by the NMDA receptor antagonist dizocilpine (MK-801). Sprague-Dawley rats (N=35) were trained on a food-motivated two-lever operant task with one lever correct after a brief visual signal (0.027-1.22 lx) for hits and the other lever correct after the absence of a signal for correct rejections. First, a dose response study of dizocilpine was conducted to determine the threshold for impairment. The rats were administered acute doses of dizocilpine (0, 12.5, 25 and 50 microg/kg, sc). The 50 microg/kg dose caused significant (p<0.0005) reduction in percent hit at the four highest signal intensities. Percent correct rejection was also significantly lowered by this dose (p<0.005). No effect was seen with 12.5 microg/kg and only minimal effect seen with 25 microg/kg. Then, nicotine dizocilpine interactions were investigated. The rats were administered acute doses of dizocilpine (0, 37.5 and 50 microg/kg, sc) and nicotine (0, 25 and 50 microg/kg, sc), alone or in combination. Percent hit was affected by nicotine and dizocilpine in a complex fashion with only the nicotinexdizocilpinexsignal intensity interaction being significant (p<0.05). Percent correct rejection showed a more straightforward effect. Percent correct rejection was significantly reduced by 50 microg/kg dizocilpine (p<0.025). The addition of 25 microg/kg of nicotine significantly (p<0.025) reversed the dizocilpine-induced reduction of correct rejection. This study shows that dizocilpine reduces signal detection accuracy in a dose-dependent fashion. Nicotine can partially counteract an aspect of this reduction by reversing the dizocilpine-induced reduction of correct rejection. PMID- 12650837 TI - Effects of histamine and cholinergic systems on memory retention of passive avoidance learning in rats. AB - In the present study, the effects of the histamine and cholinergic systems on memory retention in adult male rats were investigated. Post-training intracerebroventricular injections were carried out in all the experiments. Cholinoceptor agonist, acetylcholine (1-10 microg/rat) or nicotine (1-10 microg/rat), increased, while a cholinoceptor antagonist, scopolamine (5-20 microg/rat), decreased memory retention. The response to acetylcholine was attenuated by scopolamine. Administration of histamine (5-20 microg/rat) reduced, but the histamine H(1) receptor antagonist, pyrilamine (10-50 microg/rat), and the histamine H(2) receptor antagonist, cimetidine (1-50 microg/rat), increased memory retention in rats. The histamine receptor antagonists attenuated the response to histamine. Histamine reduced the acetylcholine- or nicotine-induced enhancement. The histamine receptor antagonists enhanced the nicotine- or acetylcholine-induced response. Histamine potentiated the inhibitory effect induced by scopolamine. It is concluded that histaminergic and cholinergic systems have opposing effects on memory retention. Also, the histaminergic system elicits an interaction with the cholinergic system in memory retention. PMID- 12650838 TI - Effects of T-82, a novel acetylcholinesterase inhibitor, on impaired learning and memory in passive avoidance task in rats. AB - Effects of 2-[2-(1-benzylpiperidin-4-yl)ethyl]-2,3-dihydro-9-methoxy-1H pyrrolo[3,4-b]quinolin-1-one hemifumarate (T-82), a new quinoline derivative, on drug- and basal forebrain lesion-induced amnesia models were examined in rats. Scopolamine (0.5 mg/kg, i.p.) and cycloheximide (1.5 mg/kg, s.c.) shortened the step-through latency in the passive avoidance task. T-82 significantly ameliorated amnesia induced by scopolamine or cycloheximide at the dose of 0.03, 0.1 and 0.3 mg/kg, p.o., and 0.3 and 1.0 mg/kg, p.o., respectively. Basal forebrain lesions with ibotenic acid shortened the step-through latency in passive avoidance task. An acute (0.1 and 0.3 mg/kg, p.o.) or subacute (0.03-0.3 mg/kg, p.o., for 7 days) treatment of T-82 significantly reversed the shortened latency. These results suggest that T-82 may ameliorate the impairment of memory induced by acetylcholinergic dysfunction. PMID- 12650839 TI - Pre-exposure to the cannabinoid receptor agonist CP 55940 enhances morphine behavioral sensitization and alters morphine self-administration in Lewis rats. AB - Three experiments examined the influence of pre-exposure to the cannabinoid receptor agonist CP 55940 ((-)-cis-3-(2-hydroxy-4-(1,1-dimethylheptyl)phenyl) trans-4-(3-hydroxypropyl)cyclohexanol) on the sensitization of morphine-induced locomotor hyperactivity and self-administration in Lewis rats. In Experiment 1, rats received daily injections of vehicle or CP 55940 (0.1 mg/kg for 7 days then 0.2 mg/kg for a further 7 days). Four weeks later, the locomotor response to morphine (10 mg/kg s.c.) was tested once per day over a 3-h period for 14 consecutive days. Rats given morphine showed hypoactivity during the first hour following morphine but hyperactivity during the second and third hours. A progressive increase in hyperactivity to morphine was seen over the 14 days of administration, which was significantly greater in rats pre-treated with CP 55940. In Experiment 2, rats were given morphine (10 mg/kg) once a day for 14 days in combination with either vehicle, CP 55940 (0.1 mg/kg) or the cannabinoid CB(1) receptor antagonist SR 141716 (N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4 dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide hydrochloride) (3 mg/kg). Both CP 55940 and SR 141716 initially inhibited the hyperactive response to morphine, but these effects gradually wore off and by the end of 14 days, hyperactivity was similar in all morphine-treated groups. When tested 3 weeks later for their response to morphine (10 mg/kg) given alone, rats previously given the morphine/CP 55940 combination, but not the SR 141716/morphine combination, showed a greater locomotor stimulation than those previously exposed to morphine only. In Experiment 3, rats were pre-exposed to CP 55940 or vehicle for 14 days and were subsequently trained to self-administer morphine intravenously (1 mg/kg per lever press) for 14 days. Rats pre-exposed to CP 55940 self-administered a significantly greater number of morphine infusions than vehicle pre-exposed rats. However, both active and inactive ('dummy') lever presses were increased by cannabinoid pre-treatment. Overall, these results suggest that cannabinoid pre exposure can lead to an exaggeration of morphine-induced hyperactivity and may alter the reinforcing effects of morphine in Lewis rats. The implications for 'gateway' theories of cannabinoid effects in humans are discussed. PMID- 12650840 TI - Resveratrol provides late-phase cardioprotection by means of a nitric oxide- and adenosine-mediated mechanism. AB - We used two experimental models to prove that resveratrol (trans-3,4',5 trihydroxystilbene) reduces cardiac ischemic-reperfusion injury by means of a nitric oxide- and adenosine-dependent mechanism. (1). ACUTE EX VIVO: resveratrol (10 microM, 10 min) infusion in Langendorff-perfused normoxic rat hearts significantly increased adenosine release and coronary flow compared with baseline. After 30-min low-flow ischemia, vasodilation, still present at reperfusion, was completely abolished by resveratrol plus adenosine antagonist 8 (p-sulfophenyl)theophylline (SPT, 50 microM) administration. (2). CHRONIC IN VIVO: rats received tap water containing 25 mg/l resveratrol for 15 days or normal water. Twenty-four hours after, their hearts were Langendorff-perfused and submitted to 60-min low-flow ischemia and reperfusion. The resveratrol-treated hearts showed better functional recovery at reperfusion and significant vasodilation, but no variation in high-energy phosphates (31P Nuclear Magnetic Resonance). N(G)-nitro-L-arginine methyl ester (L-NAME, 30 microM), a nonselective nitric oxide synthase inhibitor, or SPT (50 microM) administered for 10 min prior to the low-flow ischemia cancelled the effects. This suggests that long-term moderate resveratrol consumption could play an important role in late cardioprotective effects. PMID- 12650841 TI - Mechanisms of vasorelaxation to testosterone in the rat aorta. AB - We have investigated the role of endothelium-derived relaxing factors, K(+) channels and steroid receptors in vasorelaxation to testosterone in the rat aorta. Testosterone (1 nM-mM) caused acute concentration-dependent vasorelaxation. Both indomethacin (10 microM) and flurbiprofen (10 microM) uncovered relaxant responses to testosterone. The action of indomethacin was inhibited by endothelial removal. N(G)-nitro-L-arginine methyl ester (L-NAME, 300 microM) had no effects on testosterone-induced responses. In the presence of indomethacin, the vasorelaxant potency of testosterone was reduced by depolarization with 60 mM KCl or charybdotoxin (100 nM), but not by glibenclamide (10 microM), 4-aminopyridine (1 mM) or barium chloride (30 microM). The responses to testosterone were not inhibited by flutamide (10 microM) or mifepristone (30 microM). Pre-treatment of the aorta with testosterone (100 microM) inhibited CaCl(2)-induced contraction. In the present study, we have demonstrated that testosterone causes acute vasorelaxations, which are modulated via endothelium derived prostanoids. The responses uncovered by cyclooxygenase inhibitors are due to the activation of K(Ca) channels, while at higher concentrations, testosterone inhibits Ca(2+) influx. PMID- 12650842 TI - Type IV phosphodiesterase inhibition improves cardiac contractility in endotoxemic rats. AB - Type IV phosphodiesterase inhibitors have a potential role in treating human sepsis. We examined the cardiac performance effects of type IV phosphodiesterase inhibition in vivo, in the absence and presence of catecholamines. Rats were randomized to receive either 4-(3-Butoxy-4-methoxybenzyl)imidazolidin-2-one (Ro 20-1724) at 0 (vehicle), 2 or 10 microg/kg/min. Utilizing a left ventricular catheter to measure cardiac performance, each animal received each of the two catecholamines, epinephrine and norepinephrine, in randomized order. Rats then received intravenous endotoxin and additional infusions of catecholamines. Ro 20 1724 at 2 microg/kg/min protected cardiac contractility during endotoxemia, and at 10 microg/kg/min increased cardiac contractility and protected cardiac function during endotoxemia. Neither dose interfered with the maximal contractile response to catecholamines. Type IV phosphodiesterase inhibition with Ro 20-1724 exerts beneficial effects on cardiac performance during septicemia in an in vivo animal model. Clinical studies of type IV phosphodiesterase inhibitors in critically ill patients are indicated. PMID- 12650843 TI - Extracellular acidosis results in higher intracellular acidosis and greater contraction in spontaneously hypertensive rat aorta. AB - Acidic pH induces a contraction in aorta from spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats. The contractile response to acidic pH in SHR aorta is greater than that in WKY aorta. The purpose of this study was to investigate the correlation among extracellular pH (pH(o)), intracellular pH (pH(i)) and contraction in order to understand the exaggerated contractile response to acidic pH in SHR aorta. pH(i) measurement showed that at pH(o) 6.5, intracellular acidification was greater in SHR aorta than in WKY aorta. Decreasing pH(o) further to 6.2 in WKY aorta produced intracellular acidification close to that achieved at pH(o) 6.5 in SHR aorta, and at this level, the difference in contractile response between the two strains was also abolished. These results suggest that acidic pH(i), but not pH(o), is closely correlated with the contractile response and that the exaggerated contractile response in SHR aorta is due to a greater fall in pH(i). PMID- 12650844 TI - Mexiletine inhibits nonadrenergic noncholinergic lower oesophageal sphincter relaxation in rabbits. AB - Nonadrenergic noncholinergic (NANC) nerves are known to be nitrergic and to have an important role in the regulation of gastrointestinal motility and function. Cardiac antiarrhythmic therapy in humans is accompanied by a high incidence of gastrointestinal side-effects. We investigated the effect of mexiletine, a class Ib antiarrhythmic drug, on NANC lower oesophageal sphincter relaxation. Mexiletine concentration dependently inhibited the NANC relaxation induced by 30 mM KCl (EC(50)=4.4 x 10(-6) M); the production of 3',5'-cyclic guanosine monophosphate (cGMP) after KCl stimulation was concentration dependently decreased. The relaxation induced by the exogenous nitric oxide (NO) donor, diethylamine NONOate (10(-5) M), was not inhibited by mexiletine, and the cGMP production after diethylamine NONOate application was not altered. Mexiletine did not alter the activity of NO synthase. These findings suggest that mexiletine inhibits NANC relaxation via NO-cGMP pathway modulation, possibly by inhibiting myenteric nitrergic neurotransmission in the lower oesophageal sphincter in rabbits. PMID- 12650845 TI - Caffeine-induced contracture in oesophageal striated muscle of normotensive and hypertensive rats. AB - To elucidate whether properties of the sarcoplasmic reticulum are altered, not only in vascular smooth muscle, but also in visceral striated muscle of spontaneously hypertensive rats (SHR), caffeine-induced contractures in oesophageal striated muscle of Wistar Kyoto rats (WKY) and stroke-prone SHR (SHRSP) were compared. In both preparations, 30 mM caffeine induced a contracture with two components. The second component, which was diminished by extracellular Ca(2+) removal or Ni(2+) but not by verapamil, was much smaller in SHRSP. Both components and differences between WKY and SHRSP coincided with changes in intracellular Ca(2+). Although membrane potential was identical between these preparations, caffeine induced slight depolarization only in WKY preparations. Similar depolarization was observed with 10 mM K(+), which induced no contraction. It is suggested that the first and the second components of caffeine induced contracture were induced by Ca(2+) released from sarcoplasmic reticulum and by Ca(2+) that entered through channels activated by sarcoplasmic reticulum Ca(2+) depletion, respectively. In SHRSP preparations, Ca(2+) from the latter pathway was clearly decreased, although this change is thought not to be related to the initiation of hypertension. These results suggest that Ca(2+) handling properties of cell membrane and sarcoplasmic reticulum are generally altered in muscles of SHRSP. PMID- 12650846 TI - Edaravone protects against ischemia/reperfusion-induced oxidative damage to mitochondria in rat liver. AB - This study investigated the effects of edaravone (3-methyl-1-phenyl-2-pyrazolin-5 one, MCI-186), a potent free radical scavenger, on the prevention of mitochondrial injury induced by hepatic ischemia and reperfusion. Mature male rats were subjected to 70 min of hepatic ischemia and 2 h of reperfusion. The rats received vehicle or edaravone (10 mg/kg body weight) intravenously prior to ischemia, before reperfusion and 1 h after reperfusion. In the vehicle-treated animals, the respiratory control index, ADP/O, State 3 respiration and dinitrophenol-induced uncoupled respiration decreased markedly after ischemia/reperfusion and were restored by edaravone administration. Mitochondrial lipid peroxidation was elevated in the vehicle-treated group, which was attenuated by edaravone, while mitochondrial glutathione peroxidase activity decreased in the vehicle-treated group, which was similarly abrogated by edaravone treatment. Electron microscopic observation demonstrated that treatment with edaravone restored the ischemia/reperfusion-induced disorganization of mitochondrial structures. Edaravone protects against mitochondrial injury, which prevents mitochondrial oxidative stress and improves ischemia/reperfusion-induced hepatic energy metabolism. PMID- 12650847 TI - Mechanisms in mediating the anti-inflammatory effects of baicalin and baicalein in human leukocytes. AB - To evaluate the possible mechanisms responsible for the anti-inflammatory effects of baicalin or baicalein, phorbol-12-myristate-13-acetate (PMA)- or N-formyl methionyl-leucyl-phenylalanine (fMLP)-activated inflammatory responses of peripheral human leukocytes were studied. Both baicalin and baicalein diminished fMLP- or PMA-induced reactive oxygen intermediates production in neutrophils or monocytes. Neither baicalin nor baicalein prevented the protein kinase C (PKC) dependent assembly of the NADPH oxidase. Conversely, myeloperoxidase (MPO) activity was inhibited by baicalin or baicalein. fMLP-induced activation of leukocytes, as reflected by increased surface expression of Mac-1 (CD11b/CD18) and Mac-1-dependent neutrophil adhesion, were also inhibited by baicalin or baicalein. Furthermore, baicalein, but not baicalin, impeded fMLP- or AlF(4)(-) induced Ca(2+) influx. We conclude that impairment of reactive oxygen intermediates production, through scavenging reactive oxygen intermediates by baicalin, or antagonizing ligand-initiated Ca(2+) influx by baicalein, accounts for the inhibition of Mac-1-dependent leukocyte adhesion that confers the anti inflammatory activity of baicalin or baicalein. PMID- 12650848 TI - ttCH, a selective inhibitor of inducible nitric oxide synthase expression with antiarthritic properties. AB - In a previous work, we investigated the effects of a series of dimethoxy- and trimethoxychalcone derivatives, with various patterns of fluorination, on nitric oxide production in lipopolysaccharide-stimulated murine RAW 264.7 cells. The present study was designed to determine if 2,4,6-trimethoxy-2' trifluoromethylchalcone (ttCH) could modulate the production of nitric oxide (NO) and/or prostaglandins in vitro and in vivo. On the mouse macrophage cell line RAW 264.7, ttCH inhibited dose-dependently NO and prostaglandin E(2) production, with IC(50) in the micromolar range. This compound had no direct inhibitory effect on inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 activities. NO reduction was the consequence of inhibition of the expression of iNOS. This compound also exhibited in vivo an inhibitory behaviour on nitrite and prostaglandin E(2) levels. We have assessed the effect of ttCH in the treatment of acute and chronic inflammatory processes such as the mouse carrageenan paw oedema and the rat adjuvant-induced arthritis. The present study demonstrated that ttCH exerts acute and chronic anti-inflammatory effects that may be related with the inhibition of iNOS expression. PMID- 12650849 TI - Suppression of tumor growth by a new glycosaminoglycan isolated from the African giant snail Achatina fulica. AB - Acharan sulfate is a new type of glycosaminoglycan from the giant African snail, Achatina fulica. Acharan sulfate, which has a primary repeating disaccharide structure of alpha-D-N-acetylglucosaminyl-2-O-sulfo-alpha-L-iduronic acid, was studied as a potential antitumor agent in both in vivo and in vitro assays. The antiangiogenic activity of acharan sulfate was evaluated in the chorioallantoic membrane assay and by measuring its effect on the proliferation of calf pulmonary artery endothelial cells. In vivo, a matrigel plug assay showed that acharan sulfate suppressed basic fibroblast growth factor (bFGF)-stimulated angiogenesis and lowered the hemoglobin (Hb) content inside the plug. Acharan sulfate was administered s.c. at two doses for 15 days to C57BL/6 mice implanted with murine Lewis lung carcinoma in the back. It was also administered i.p. to ICR mice bearing sarcoma 180 at a dose of 30 mg/kg. Subcutaneous injection of acharan sulfate at doses of 10 and 30 mg/kg decreased tumor weight and tumor volume by 40% without toxicity or resistance. Intraperitoneal injection of acharan sulfate also decreased tumor weight and volume by 40% in sarcoma 180-bearing mice. These results suggest that the antitumor activity of acharan sulfate may be related to the inhibition of angiogenesis. PMID- 12650850 TI - Reversal of morphine antinociceptive tolerance by acute spinal inhibition of Ca(2+)/calmodulin-dependent protein kinase II. AB - It has been reported that Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) can modulate opioid tolerance via its action on learning and memory. In this study, we examine if CaMKII can directly affect opioid tolerance. We found that spinal CaMKII activity was increased in rats tolerant to morphine. In these rats, acute spinal administration of 2-[N-(2-hydroxyethyl)]-N-(4 methoxybenzenesulfonyl)]amino-N-(4-chlorocinnamyl)-N-methylbenzylamine) (KN93), a CaMKII inhibitor, was able to reverse the already-established antinociceptive tolerance. These results suggest that CaMKII may directly promote opioid tolerance. PMID- 12650852 TI - 5-HT2A and 5-HT2C receptors and their atypical regulation properties. AB - The 5-HT(2A) and 5-HT(2C) receptors belong to the G-protein-coupled receptor (GPCR) superfamily. GPCRs transduce extracellular signals to the interior of cells through their interaction with G-proteins. The 5-HT(2A) and 5-HT(2C) receptors mediate effects of a large variety of compounds affecting depression, schizophrenia, anxiety, hallucinations, dysthymia, sleep patterns, feeding behaviour and neuro-endocrine functions. Binding of such compounds to either 5 HT(2) receptor subtype induces processes that regulate receptor sensitivity. In contrast to most other receptors, chronic blockade of 5-HT(2A) and 5-HT(2C) receptors leads not to an up- but to a (paradoxical) down-regulation. This review deals with published data involving such non-classical regulation of 5-HT(2A) and 5-HT(2C) receptors obtained from in vivo and in vitro studies. The underlying regulatory processes of the agonist-induced regulation of 5-HT(2A) and 5-HT(2C) receptors, commonly thought to be desensitisation and resensitisation, are discussed. The atypical down-regulation of both 5-HT(2) receptor subtypes by antidepressants, antipsychotics and 5-HT(2) antagonists is reviewed. The possible mechanisms of this paradoxical down-regulation are discussed, and a new hypothesis on possible heterologous regulation of 5-HT(2A) receptors is proposed. PMID- 12650853 TI - Salvia miltiorrhiza attenuates the changes in contraction and intracellular calcium induced by anoxia and reoxygenation in rat cardiomyocytes. AB - The aim of the present study is to investigate the effect of Salvia miltiorrhiza (SM) on contraction and the intracellular calcium of isolated ventricular myocytes during normoxia or anoxia and reoxygenation using a video tracking system and spectrofluorometry. Cardiac ventricular myocytes were isolated enzymatically by collagenase and exposed to 5 min of anoxia followed by 10 min of reoxygenation. SM (1-9 g/L) depressed both contraction and the [Ca(2+)](i) transient in a dose-dependent manner. SM did not affect the diastolic calcium level and the sarcolemmal Ca(2+) channel of myocytes but decreased the caffeine induced calcium release. During anoxia, the +/-dL/dtmax, amplitudes of contraction (dL) of cell contraction and [Ca(2+)](i) transients were decreased, while the diastolic calcium level was increased. None of the parameters returned to the pre-anoxia level during reoxygenaton. However, SM (3 g/L) did attenuate the changes in cell contraction and intracellular calcium induced by anoxia and reoxygenation. It is concluded that SM has different effects on normoxic and anoxic cardiomyocytes. The SM-induced reduction of changes in contraction and intracellular calcium induced by anoxia/reoxygenation indicates that SM may be beneficial for cardiac tissue in recovery of mechanical function and intracellular calcium homeostasis. PMID- 12650854 TI - Tetramethylpyrazine scavenges superoxide anion and decreases nitric oxide production in human polymorphonuclear leukocytes. AB - Tetramethylpyrazine is one of the active ingredients of the Chinese herb Ligusticum wallichii Franchat. By electron spin resonance spin trapping methods, effects of tetramethylpyrazine on superoxide anion and nitric oxide generated by human polymorphonuclear leukocytes were studied. During the respiratory burst of polymorphonuclear leukocytes induced by N-formylmethionyl-leucyl-phenylalanine, tetramethylpyrazine scavenges superoxide anion dose-dependently, and decreases the production of nitric oxide significantly, but shows no influence on oxygen consumption. These results suggest that the effective protection of tetramethylpyrazine against ischemic brain injury might be due to its scavenging of reactive oxygen species and regulation on nitric oxide production, and consequent prevention of peroxynitrite formation. PMID- 12650855 TI - GH-RH antagonist (MZ-4-71) inhibits VEGF secretion and proliferation of murine endothelial cells. AB - Angiogenesis plays a key role in solid tumor formation, invasiveness and metastasis. Vascular endothelial growth factor (VEGF) is an endothelial cell specific mitogen that is necessary in the process of neovascularisation. Antagonists of growth hormone-releasing hormone (GH-RH) have been shown to suppress both in vivo and in vitro growth and metastasis of many human cancer cell lines. The mechanisms that mediate the antitumorigenic actions of these antagonists involve direct and indirect pathways, but are not completely elucidated. We have examined the effect of GH-RH antagonist MZ-4-71 on proliferation activity and VEGF release from cultured murine endothelial cells HECa10 in vitro. MZ-4-71 at 10(-8) to 10(-6) M concentrations inhibited the proliferative activity of cultured cells and suppressed the release of VEGF into supernatants of 72 h endothelial cell cultures. To our knowledge this is the first study reporting antiangiogenic properties of GH-RH antagonists. PMID- 12650856 TI - Endogenous gamma-hydroxybutyric acid is in the rat, mouse and human gastrointestinal tract. AB - By using Gas Chromatography-Mass Spectrometry high concentrations of endogenous gamma-hydroxybutyric acid (GHB) have been demonstrated in the rat and mouse gastrointestinal tract, including stomach, small intestine and colon-rectum. GHB concentrations were many folds higher than those present in the brain. High GHB concentrations have been also found in the human operatory specimen of sigmoid colon. Since GHB administration has been found to modify gastrointestinal motility via GABA(B) receptors, the present results suggest that endogenous GHB might be involved in the GABA(B) receptor-mediated control of gastrointestinal function. PMID- 12650857 TI - Combined effects of complex magnetic fields and agmatine for contextual fear learning deficits in rats. AB - Acute post-training exposures to weak intensity theta-burst stimulation (TBS) patterned complex magnetic fields attenuated the magnitude of conditioned fear learning for contextual stimuli. A similar learning impairment was evoked in a linear and dose-dependent manner by pre-conditioning injections of the polyamine agmatine. The present study examined the hypothesis that whole-body applications of the TBS complex magnetic field pattern when co-administered with systemic agmatine treatment may combine to evoke impairments in contextual fear learning. Within minutes of 4 mg/kg agmatine injections, male Wistar rats were fear conditioned to contextual stimuli and immediately exposed for 30 min to the TBS patterned complex magnetic field or to sham conditions. TBS patterned complex magnetic field treatment was found to linearly summate with the contextual fear learning impairment evoked by agmatine treatment alone. Furthermore, we report for sham-treated rats, but not rats exposed to the synthetic magnetic field pattern, that the magnitude of learned fear decreased and the amount of variability in learning increased, as the K-index (a measure of change in intensity of the time-varying ambient geomagnetic field) increased during the 3 hr intervals over which conditioning and testing sessions were conducted. PMID- 12650858 TI - Effect of berberine on catecholamine levels in rats with experimental cardiac hypertrophy. AB - The aim of this study is to investigate the effect of berberine on catecholamine level (adrenaline and noradrenaline) in rats with experimental cardiac hypertrophy. Cardiac hypertrophy(CH) was induced by suprarenal abdominal aorta constriction, and the drugs were administered for 8 weeks starting from 4 weeks after surgery. The degree of cardiac hypertrophy was determined by heart and left ventricular weight. The level of adrenaline(AD) and noradrenaline(NA) was detected by HPLC. The data showed that in the CH model rats, the level of plasma and left ventricular tissue AD, and the level of NA in plasma were higher than that of the age-matched controls(indicating increased "total" sympathetic activity). The level of NA in left ventricular tissue of CH model rats was however lower than the age-matched controls. Berberine and captopril showed significant effect on inhibiting the development of cardiac hypertrophy. Berberine decreased plasma NA level and the AD level both in plasma and left ventricular tissue, but had no effect on improving the cardiac NA depletion. Captopril showed significant effect on increasing the depleted cardiac NA and in reducing the elevated plasma NA level. These findings show the efficacy of berberine on modulating the sympathetic nervous activity of rats with experimental cardiac hypertrophy, and reflect the therapeutic potentials of berberine in patients with cardiac hypertrophy and chronic heart failure. PMID- 12650859 TI - Oxidative stress and altered antioxidant defenses in children with acute exacerbation of atopic dermatitis. AB - The underlying mechanisms of skin inflammation in atopic dermatitis (AD) are not completely understood. The purpose of the present study was to examine the involvement of oxidative stress and antioxidant defenses in children with acute exacerbation of AD. We studied 13 children who were hospitalized for acute exacerbation of AD with purulent skin infection by Staphylococcal aureus (age, 1.5 to 10.0 years), and 28 age-matched healthy subjects (controls). Urine samples obtained from the patients on admission, on 2nd and 7th-9th hospital days, as well as from the controls were analyzed for 8-hydroxy-2'-deoxyguanosine (8-OHdG) (a marker of oxidative DNA damage), acrolein-lysine adducts (a marker of lipid peroxidation), bilirubin oxidative metabolites (BOM) (a marker of antioxidant activity of bilirubin under oxidative stress) and nitrite/nitrate (NO(x)(-)) (a marker of endogenous nitric oxide production). Of these, urinary concentrations of 8-OHdG, acrolein-lysine adducts and BOM, but not NO(x)(-), were significantly higher in AD children on admission than those in control subjects. Response to treatment was associated with significant falls in the concentrations of 8-OHdG and acrolein-lysine adducts. Urinary concentrations of acrolein-lysine adducts, but not 8-OHdG, were still significantly higher in AD patients on the 7th-9th hospital day relative to the control. Urinary BOM remained almost constant and significantly high in AD children during hospitalization. Our findings indicate that oxidative stress and altered antioxidant defenses are involved in the pathophysiology of acute exacerbation of AD, and that suppression of oxidative stress might be a potentially useful strategy for the treatment of AD. PMID- 12650860 TI - The role of circulating ghrelin in growth hormone (GH) secretion in freely moving male rats. AB - To examine the physiological significance of plasma ghrelin in generating pulsatile growth hormone (GH) secretion in rats, plasma GH and ghrelin levels were determined in freely moving male rats. Plasma GH was pulsatilely secreted as reported previously. Plasma ghrelin levels were measured by both N-RIA recognizing the active form of ghrelin and C-RIA determining total amount of ghrelin. Mean +/- SE plasma ghrelin levels determined by N-RIA and C-RIA were 21.6 +/- 8.5 and 315.5 +/- 67.5 pM, respectively, during peak periods when plasma GH levels were greater than 100 ng / ml. During trough periods when plasma GH levels were less than 10 ng / ml, they were 16.5 +/- 4.5 and 342.1 +/- 29.8 pM, respectively. There were no significant differences in plasma ghrelin levels between two periods. Next, effect of a GH secretagogue antagonist, [D-Lys-3]-GHRP 6, on plasma GH profiles was examined. There were no significant differences in both peak GH levels and area under the curves of GH (AUCs) between [D-Lys-3]-GHRP 6-treated and control rats. These findings suggest circulating ghrelin in peripheral blood does not play a role in generating pulsatile GH secretion in freely moving male rats. PMID- 12650861 TI - Incorporation of DNA chip technology to the simulation and validation of flux analysis in yeast diauxic growth. AB - We incorporated gene expression information from cDNA microarray into flux analysis to simulate yeast diauxic growth. Expression ratios of both growth phases were applied to assign the split ratio at glyoxylate shunt during simulation, in which the equation was mathematically unsolvable due to the singularity and artificial split ratios, which were traditionally introduced without biological evidence. In addition, the directionality of microarray dataset was used as a further constraint during simulation. Metabolic fluxes obtained by this modified approach are in general consistent with microarray analysis. However, discrepancies occurred when the quantity of fluxes was compared, probably due to the substantial reduction of substrates at phase II in which the increase in the enzymatic levels was not proportional to the increase of substrate flow, as would be predicted from microarray dataset. The modified flux analysis might have brought a new approach to investigate other cellular pathways. PMID- 12650862 TI - Nongenomic mechanism of glucocorticoid inhibition of bradykinin-induced calcium influx in PC12 cells: possible involvement of protein kinase C. AB - Many stimulants, including bradykinin (BK), can induce increase in [Ca(2+)](i) in PC12 cells. Bradykinin induces an increase in [Ca(2+)](i) via intracellular Ca(2+) release and extracellular Ca(2+) influx through the transduction of G protein, but not through voltage-sensitive calcium channels. In this experiment, We analyzed how corticosterone (Cort) influences BK-induced intracellular Ca(2+) release and extracellular Ca(2+) influx, and further studied the mechanism of glucocorticoid's action. To dissociate the intracellular Ca(2+) release and extracellular Ca(2+) influx induced by BK, the Ca(2+)-free/Ca(2+)- reintroduction protocol was used. The results were as follows: (1) The Ca(2+) influx induced by BK could be rapidly inhibited by Cort, but intracellular Ca(2+) release could not be affected significantly. (2) The inhibitory effect of Cort-BSA (BSA -conjugated Cort) on Ca(2+) influx induced by BK was the same as the effect of free Cort. (3) Protein kinase C (PKC) activator (phorbol 12-myristate 13-acetate) could mimic and PKC inhibitor Go6976 could reverse the inhibitory effect of Cort. (4) There was no inhibitory effect of Cort on Ca(2+) influx induced by BK when pretreated with pertussis toxin. The results suggested, for the first time, that Cort might act via a putative membrane receptor and inhibit the Ca(2+) influx induced by BK through the pertussis toxin -sensitive G protein-PKC pathway. PMID- 12650863 TI - Cycloamphilectenes, a new type of potent marine diterpenes: inhibition of nitric oxide production in murine macrophages. AB - The inhibitory effect of a series of 6 cycloamphilectenes, novel marine diterpenes based on amphilectene skeletons and isolated from the Vanuatu sponge Axinella sp., on NO, PGE(2) and TNFalpha production in murine peritoneal macrophages was studied. These compounds reduced potently nitric oxide production in a concentration-dependent manner with IC(50) values in the submicromolar range (0.1-4.3 microM). Studies on intact cells and Western blot analysis showed that the more potent cycloamphilectenes reduced the expression of inducible nitric oxide synthase without affecting cyclo-oxygenase-2 expression. Among them cycloamphilectene 2, the unique compound bearing an exocyclic methylene group, was able to reduce NO production without affecting TNFalpha release. Cycloamphilectene 2, which is an inhibitor of the nuclear factor-kB pathway, exhibited topical anti-inflammatory activity. PMID- 12650864 TI - Sodium and the heart: a hidden key factor in cardiac regulation. PMID- 12650866 TI - [Na+]i handling in the failing human heart. AB - Proper contractile function of the heart depends on intact excitation-contraction processes and ion homeostasis of the myocytes. The Ca2+ ion activates contraction through its binding to troponin C. However, Ca2+ homeostasis is tightly linked to Na+ regulation because the primary mechanism for Ca2+ efflux in cardiac myocytes is via electrogenic Na+/Ca2+-exchange. While altered Ca2+-homeostasis has been demonstrated in animal models of heart failure and failing human cardiac tissue, the role of dysfunctional Na+ handling processes in altered excitation contraction coupling remains obscure. Furthermore, altered Na+ handling has been implicated in a wide range of cellular processes, such as regulation of membrane potential, pH, and growth. This review will discuss (1) the evidence for altered [Na+]i homeostasis in the failing human heart, (2) how alterations in the Na+ electrochemical gradient can influence Ca2+ handling, contractile function, and a number of other cellular processes, and (3) the potential defects in Na+ channels and transporters that may underlie altered [Na+]i in the failing human heart. PMID- 12650867 TI - Intracellular Na in animal models of hypertrophy and heart failure: contractile function and arrhythmogenesis. PMID- 12650868 TI - Intracellular Na+ regulation in cardiac myocytes. AB - Intracellular [Na+] ([Na+]i) is regulated in cardiac myocytes by a balance of Na+ influx and efflux mechanisms. In the normal cell there is a large steady state electrochemical gradient favoring Na+ influx. This potential energy is used by numerous transport mechanisms, including Na+ channels and transporters which couple Na+ influx to either co- or counter-transport of other ions and solutes. Six sarcolemmal Na+ influx pathways are discussed in relatively quantitative terms: Na+ channels, Na+/Ca2+ exchange, Na+/H+ exchange, Na+/Mg2+ exchange, Na+/HCO3- cotransport and Na+/K+/2Cl- cotransport. Under normal conditions Na+/Ca2+ exchange and Na+ channels are the dominant Na+ influx pathways, but other transporters may become increasingly important during altered conditions (e.g. acidosis or cell volume stress). Mitochondria also exhibit Na+/Ca2+ antiporter and Na+/H+ exchange activity that are important in mitochondrial function. These coupled fluxes of Na+ with Ca2+, H+ and HCO3- make the detailed understanding of [Na+]i regulation pivotal to the understanding of both cardiac excitation-contraction coupling and pH regulation. The Na+/K+-ATPase is the main route for Na+ extrusion from cells and [Na+]i is a primary regulator under physiological conditions. [Na+]i is higher in rat than rabbit ventricular myocytes and the reason appears to be higher Na+ influx in rat with a consequent rise in Na+/K+-ATPase activity (rather than lower Na+/K+-ATPase function in rat). This has direct functional consequences. There may also be subcellular [Na+]i gradients locally in ventricular myocytes and this may also have important functional implications. Thus, the balance of Na+ fluxes in heart cells may be complex, but myocyte Na+ regulation is functionally important and merits focused attention as in this issue. PMID- 12650869 TI - The Na, K-ATPase in the failing human heart. AB - The Na, K-ATPase consists of alpha- and beta-subunits and actively transports Na out and K into the myocyte. It is the receptor for cardiac glycosides exerting its positive inotropic effect by inhibiting enzyme activity, decreasing the driving force for the Na/Ca-exchange and increasing cellular content and release of Ca during depolarization. The specific binding capacity for cardiac glycosides is utilized as a tool for Na, K-ATPase quantification with high accuracy and precision. In treatment of patients with heart failure cardiac glycosides improve symptoms and reduce the need for hospitalization without affecting mortality. In endomyocardial biopsies from patients with compromised cardiac function total Na, K-ATPase concentration is decreased by approximately 40% and a correlation between decrease in heart function and decrease in Na, K-ATPase concentration exists. At the subunit level, the alpha1-, alpha3- and beta1-proteins are reduced in human heart failure. During digitalization approximately 30% of remaining Na, K-pumps are occupied by digoxin. Thus, a total of not less than half the Na, K pumps may be out of function in the myocardium of digitalised heart failure patients. It is still a matter of debate whether a digitalis-like factor exists. There is a pressing need for the identification of its precise chemical structure, properties and quantitative relation to the Na, K-ATPase. It is recommended that cardiac glycosides are prescribed to heart failure patients who are still having heart failure symptoms after institution of mortality reducing therapy. Cardiac glycoside treatment is still the only safe inotropic drug for oral use that improves hemodynamics in patients with compromised cardiac function. PMID- 12650870 TI - Relevance of Na+-Ca2+ exchange in heart failure. PMID- 12650871 TI - Role of the cardiac Na+/H+ exchanger during ischemia and reperfusion. AB - The coupled exchanger theory describes one of the central mechanisms of damage in the ischemic heart. The theory proposes that anaerobic glycolysis produces lactate and protons and that the protons can leave the cardiac cell on the cardiac Na+/H+ exchanger (NHE1). The subsequent rise in [Na+]i stimulates the cardiac Na+/Ca2+ exchanger (NCX) and results in an increase in [Ca2+]i which promotes myocardial cell damage. Although the general features of this theory are widely accepted, there is dispute about some aspects, specifically whether the NHE1 remains active during ischemia or not. We review the evidence on this issue and conclude that NHE1 is substantially inhibited during ischemia. This issue is central to the design of a clinical trial of NHE1 inhibitors in the treatment of human cardiac ischemia and the existing clinical trials are considered in this light. PMID- 12650872 TI - Regulatory effects of G protein-coupled receptors on cardiac sarcolemmal Na+/H+ exchanger activity: signalling and significance. AB - In cardiac myocytes, sarcolemmal Na+/H+ exchanger (NHE) activity is subject to regulation by a variety of G protein-coupled receptor (GPCR) systems. This regulation usually manifests as an increase in NHE activity (e.g. in response to the stimulation of alpha1-adrenergic, angiotensin AT1, endothelin and thrombin receptors), although some GPCR systems have been shown to inhibit sarcolemmal NHE activity (e.g. beta1-adrenergic receptors) or to attenuate its stimulation by other ligands (e.g. adenosine A1 and angiotensin AT2 receptors). The pertinent molecular signalling mechanisms are only now beginning to be unravelled, with the extracellular signal regulated kinase/ribosomal S6 kinase pathway and the protein kinase C pathway both appearing to play critical roles in the stimulation of sarcolemmal NHE activity. GPCR-mediated regulation of sarcolemmal NHE activity is likely to play significant roles in modulating myocardial function in both physiological and pathophysiological conditions. These roles include the regulation of (1) myocardial pH(i) and contractility, (2) myocardial susceptibility to injury and dysfunction during ischaemia and reperfusion, and (3) myocardial hypertrophy in response to neurohormonal and mechanical stimuli. Greater understanding of the pertinent molecular signalling mechanisms distal to GPCR stimulation may reveal novel targets for therapeutic manipulation. PMID- 12650873 TI - Stretch-elicited Na+/H+ exchanger activation: the autocrine/paracrine loop and its mechanical counterpart. AB - The stretch of the cardiac muscle is immediately followed by an increase in the contraction strength after which occurs a slow force increase (SFR) that takes several minutes to fully develop. The SFR was detected in a wide variety of experimental preparations including isolated myocytes, papillary muscles and/or trabeculae, left ventricle strips of failing human myocardium, in vitro isovolumic and in vivo volume-loaded hearts. It was established that the initial increase in force is due to an increase in myofilament Ca2+ responsiveness, whereas the SFR results from an increase in the Ca2+ transient. However, the mechanism(s) for this increase in the Ca2+ transient has remained undefined until the proposal of Na+/H+ exchanger (NHE) activation by stretch. Studies in multicellular cardiac muscle preparations from cat, rabbit, rat and failing human heart have shown evidence that the stretch induces a rise in intracellular Na+ ([Na+]i) through NHE activation, which subsequently leads to an increase in Ca2+ transient via reverse-mode Na+/Ca2+ (NCX) exchange. These experimental data agree with a theoretical ionic model of cardiomyocytes that predicted an increased Na+ influx and a concurrent increase in Ca2+ entry through NCX as the cause of the SFR to muscle stretch. However, there are aspects that await definitive demonstration, and perhaps subjected to species-related differences like the possibility of an autocrine/paracrine loop involving angiotensin II and endothelin as the underlying mechanism for stretch-induced NHE activation leading to the rise in [Na+]i and reverse-mode NCX. PMID- 12650874 TI - Genetic control of sodium channel function. AB - Sodium ion (Na) influx through cardiac Na channels triggers the action potential in cells of the working myocardium and the specialized conduction system. Na channels thus act as key molecular determinants of cardiac excitability and impulse propagation. Na channel dysfunction may cause life-threatening arrhythmias. Here, we review the ways in which Na channel function can be aberrant due to genetic changes. We discuss how biophysical studies of mutant Na channels combined with precise clinical phenotyping may improve our understanding of Na channel function in health and disease and may be useful as a model from which to derive improved treatment strategies for common disease. PMID- 12650875 TI - Calcium entry via Na/Ca exchange during the action potential directly contributes to contraction of failing human ventricular myocytes. AB - Prolongation of the Ca2+ transient and action potential (AP) durations are two characteristic changes in myocyte physiology in the failing human heart. The hypothesis of this study is that Ca2+ influx via reverse mode Na+/Ca2+ exchanger (NCX) or via L-type Ca2+ channels directly activates contraction in failing human myocytes while in normal myocytes this Ca2+ is transported into the sarcoplasmic reticulum (SR) to regulate SR Ca2+ stores. METHODS: Myocytes were isolated from failing human (n=6), nonfailing human (n=3) and normal feline hearts (n=9) and whole cell current and voltage clamp techniques were used to evoke and increase the duration of APs (0.5 Hz, 37 degrees C). Cyclopiazonic acid (CPA 10(-6) M), nifedipine (NIF;10(-6) M) and KB-R 7943 (KB-R; 3x10(-6) M) were used to reduce SR Ca2+ uptake, Ca2+ influx via the L-type Ca2+ current and reverse mode NCX, respectively. [Na+)i was changed by dialyzing myocytes with 0, 10 and 20 mM Na(+) pipette solutions. RESULTS: Prolongation of the AP duration caused an immediate prolongation of contraction and Ca2+ transient durations in failing myocytes. The first beat after the prolonged AP was potentiated by 21+/-5 and 27+/-5% in nonfailing human and normal feline myocytes, respectively (P<0.05), but there was no significant effect in failing human myocytes (+5+/-4% vs. steady state). CPA blunted the potentiation of the first beat after AP prolongation in normal feline and nonfailing human myocytes, mimicking the failing phenotype. NIF reduced steady state contraction in feline myocytes but the potentiation of the first beat after AP prolongation was unaltered (21+/-3% vs. base, P<0.05). KB-R reduced basal contractility and abolished the potentiation of the first beat after AP prolongation (2+/-1% vs. steady state). Increasing [Na+]i shortened AP, Ca2+ transient and contraction durations and increased steady state and post AP prolongation contractions. Dialysis with 0 Na+ eliminated these effects. CONCLUSIONS: Ca2+ enters both normal and failing cardiac myocytes during the late portion of the AP plateau via reverse mode NCX. In (normal) myocytes with good SR function, this Ca(2+) influx helps maintain and regulate SR Ca2+ load. In (failing) human myocytes with poor SR function this Ca2+ influx directly contributes to contraction. These studies suggest that the Ca2+ transient of the failing human ventricular myocytes has a higher than normal reliance on Ca2+ influx via the reverse mode of the NCX during the terminal phases of the AP. PMID- 12650876 TI - [Na+]i and the driving force of the Na+/Ca2+-exchanger in heart failure. AB - OBJECTIVE: Diastolic calcium is increased in myocytes from failing hearts despite up-regulation of the principal calcium extruding mechanism the Na+/Ca2+-exchanger (NCX). We hypothesize that increased diastolic calcium ([Ca2+]i) is secondary to increased cytosolic sodium ([Na+]i) and decreased driving force of NCX (DeltaG(exch)). METHODS: The stimulation rate dependence of simultaneously measured cytosolic sodium ([Na+]i), calcium transients ([Ca2+]i) and action potentials were determined with SBFI, indo-1 and the perforated patch technique in midmural left ventricular myocytes isolated from rabbits with pressure and volume overload induced heart failure (HF) and in age matched controls. Dynamic changes of DeltaG(exch) were calculated. RESULTS: With increasing stimulation frequency, 0.2-3 Hz (all data HF versus control): [Na+]i increased (6.4 to 10.8 versus 3.8 to 6.4 mmol/l), diastolic [Ca2+]i increased (142 to 219 versus 47 to 98 nmol/l), calcium transient amplitude decreased in HF (300 to 250 nmol/l) but increased in control (201 to 479 nmol/l), action potential duration (APD90) decreased (380 to 260 versus 325 to 205 ms) and time averaged DeltaG(exch) decreased (6.8 to 2.8 versus 8.7 to 6.4 kJ/mol. With increasing stimulation rate the forward mode time integral of DeltaG(exch) decreased in HF by about 30%, the reversed mode time integral increased about ninefold and the duration of reversed mode operation more than sixfold relative to control. CONCLUSIONS: [Na+]i is increased in HF and the driving force of NCX is decreased. NCX exerts thermodynamic control over diastolic calcium. Disturbed diastolic calcium handling in HF is due to decreased forward mode DeltaG(exch) secondary to increased [Na+]i and prolongation of the action potential. Enhanced reversed mode DeltaG(exch) may account for increased contribution of NCX to e-c coupling in HF. PMID- 12650877 TI - The functional effect of adenoviral Na+/Ca2+ exchanger overexpression in rabbit myocytes depends on the activity of the Na+/K+-ATPase. AB - OBJECTIVES: The functional consequences of Na+/Ca2+ exchanger (NCX) overexpression in heart failure have been controversially discussed. NCX function strongly depends on intracellular sodium which has been shown to be increased in heart failure. METHODS AND RESULTS: We investigated the Na+/K+-ATPase (NKA) inhibitor ouabain (0.5-16 micromol/l) in electrically stimulated, isotonically contracting adult rabbit cardiocytes overexpressing NCX after adenoviral gene transfer (Ad-NCX-GFP, 48 h culture time). Myocytes transfected with adenovirus encoding for green fluorescent protein (Ad-GFP) served as a control. Contractions were analyzed by video-edge detection. In the Ad-NCX-GFP group, the maximum inotropic response was significantly reduced by 50.7% (P<0.05). This was a result of an enhanced susceptibility to contracture after exposure to the drug (median concentration (25-75%): 4 (4-8) vs. 8 (6-16) micromol/l, P<0.05). When analyzing relaxation before contracture, the maximum relaxation velocity was reduced (0.15+/-0.04 vs. 0.27+/-0.04 microm/s, P<0.05) and the time from peak shortening to 90% of relaxation was increased (298+/-39 vs. 185+/-15 ms, P<0.05). No differences in systolic and diastolic parameters were observed with the Na+ channel modulator BDF9198 (1 micromol/l). CONCLUSIONS: Inhibition of NKA by ouabain induces a combined diastolic and systolic dysfunction in NCX overexpressing rabbit myocytes. This may be the consequence of cytoplasmic Ca2+ overload due to inhibition of forward mode or induction of reverse mode Na+/Ca2+ exchange. In end-stage failing human myocardium and during digitalis treatment this mechanism may be of major importance. PMID- 12650878 TI - Contribution of Na+/H+ exchange to Na+ overload in the ischemic hypertrophied hyperthyroid rat heart. AB - OBJECTIVE: The mechanisms responsible for intracellular ion homeostasis in ischemic hypertrophied myocardium are not fully known. Moderately hypertrophied hyperthyroid hearts (T3) are characterized by the bioenergetic changes and increased Na+/H+ exchange (NHE) activity comparable with those observed in humans and experimental models of hypertrophy. Here we test the hypothesis whether NHE inhibition in T3 heart improves ion homeostasis during ischemia and contractile function during recovery. METHODS: We compared intracellular H+ (H+i) and Na+ (Na+i) accumulations during 28 min global ischemia in isolated perfused T3 and euthyroid (EUT) rat hearts with and without NHE inhibition by using 31P and 23Na NMR. Heart function was measured during control perfusion and 30 min following ischemic insult. RESULTS: In T3 hearts ischemia caused: (1) faster and greater Na+i accumulation (534+/-25% of preischemic level versus 316+/-22% in EUT, P<0.001); (2) lower acidification (pH(i) 6.66+/-0.66 versus 6.12+/-0.12 in EUT, P<0.001); and (3) faster hydrolysis of ATP. NHE inhibition (amiloride 1 mM) in T3 hearts lead to: (1) delayed and lower Na+i accumulation by 35+/-5%; (2) faster and greater acidification (pH(i) 6.45+/-0.15, P<0.05); (3) delayed ATP degradation; and (4) improved heart function during recovery. When NHE was inhibited, all T3 hearts (n=11) recovered 68+/-10% of their preischemic rate pressure product (RPP), while only two untreated T3 hearts (from 11) recovered approximately 40% of preischemic RPP. CONCLUSIONS: These data suggest that NHE inhibition could be useful intervention for the prevention of ischemic/reperfusion cell injury and could improve the function of the hypertrophied heart after acute ischemia. PMID- 12650879 TI - Increased Na+/H+-exchange activity is the cause of increased [Na+]i and underlies disturbed calcium handling in the rabbit pressure and volume overload heart failure model. AB - OBJECTIVE: Cytosolic sodium ([Na+]i) is increased in heart failure (HF). We hypothesize that up-regulation of Na+/H+-exchanger (NHE) in heart failure is causal to the increase of [Na+]i and underlies disturbance of cytosolic calcium ([Ca2+]i) handling. METHODS: Heart failure was induced in rabbits by combined volume and pressure overload. Age-matched animals served as control. [Na+]i, cytosolic calcium [Ca2+]i and cytosolic pH (pH(i)) were measured in isolated left ventricular midmural myocytes with SBFI, indo-1 and SNARF. SR calcium content was measured as the response of [Ca2+]i to rapid cooling (RC). Calcium after transients were elicited by cessation of rapid stimulation (3 Hz) in the presence of 100 nmol/l noradrenalin. NHE and Na+/K+-ATPase activity were inhibited with 10 micromol/l cariporide and 100 micromol/l ouabain, respectively. RESULTS: At all stimulation rates (0-3 Hz) [Na+]i and diastolic [Ca2+]i were significantly higher in HF than in control. With increasing frequency [Na+]i and diastolic [Ca2+]i progressively increased in HF and control, and the calcium transient amplitude (measured as total calcium released from SR) decreased in HF and increased in control. In HF (at 2 Hz), SR calcium content was reduced by 40% and the calcium gradient across the SR membrane by 60%. Fractional systolic SR calcium release was 90% in HF and 60% in control. In HF the rate of pH(i) recovery following acid loading was much faster at all pH(i) and NHE dependent sodium influx was almost twice as high as in control. In HF cariporide (10 micromol/l, 5 min) reduced [Na+]i and end diastolic [Ca2+]i to almost control values, and reversed the relation between calcium transient amplitude and stimulation rate from negative to positive. It increased SR calcium content and SR membrane gradient and decreased fractional systolic SR depletion to 60%. Cariporide greatly reduced the susceptibility to develop calcium after-transients. In control animals, cariporide had only minor effects on all these parameters. Increase of [Na+]i with ouabain in control myocytes induced abnormal calcium handling as found in HF. CONCLUSIONS: In HF up-regulation of NHE activity is causal to increased [Na+]i and secondarily to disturbed diastolic, systolic and SR calcium handling. Specific inhibition of NHE partly normalized [Na+]i, end diastolic [Ca2+]i, and SR calcium handling and reduced the incidence of calcium after-transients. Chronic treatment with specific NHE inhibitors may provide a useful future therapeutic option in treatment of developing hypertrophy and heart failure. PMID- 12650880 TI - Is there a transient rise in sub-sarcolemmal Na and activation of Na/K pump current following activation of I(Na) in ventricular myocardium? AB - OBJECTIVE: The primary aim of this study was to investigate whether activation of Na influx via voltage-gated Na channels can elevate sub-sarcolemmal ('fuzzy space') [Na] and transiently activate Na/K pump current (I(p)). METHODS AND RESULTS: Initially, Na/K pump activity was characterised in whole-cell voltage clamped single guinea-pig ventricular myocytes. I(p) was activated by intracellular Na with a K(m) of 15.5 mM and a Hill coefficient of 1.7. Extracellular K activated I(p) with a K(m) of 1.6 mM. In these experiments, a finite ouabain-sensitive I(p) was measured when the pipette [Na] was zero. This suggests that there is an accumulation of Na in a sub-sarcolemmal space that is not in equilibrium with the bulk cytosol (which is assumed to be efficiently dialysed by the low-resistance patch-pipettes used). Such a sub-sarcolemmal Na gradient was observed in separate experiments in intact rabbit papillary muscles using electron probe X-ray microanalysis. In these studies, a fuzzy-space of limited Na diffusion was observed 100-200 nm below the sarcolemmal membrane. This sub-sarcolemmal Na gradient was similar whether muscles were frozen at peak systole or end-diastole suggesting that the fuzzy-space Na does not change over the course of the contractile cycle. This was further investigated in isolated guinea pig myocytes where evidence for a transient activation of I(p) was sought immediately after the activation of voltage-gated Na channels. A single clamp step from -80 to 0 mV activated Na influx but, in the 10-2000 ms immediately following the initial Na influx no evidence for a transient activation of I(p) was observed. Similarly, no activation of I(p) could be detected immediately following a train of 20 rapid (5-Hz) pulses designed to maximise Na influx. CONCLUSIONS: These studies provide evidence for the existence of a maintained sub sarcolemmal elevation of [Na] in ventricular myocardium; however, this fuzzy space [Na] did not change immediately after the activation of Na influx via voltage-gated Na channels or throughout the contractile cycle. PMID- 12650881 TI - Increased Na+ concentration and altered Na/K pump activity in hypertrophied canine ventricular cells. AB - OBJECTIVE: To investigate whether hypertrophy in the dog with chronic atrioventricular block (CAVB) alters [Na+]i and Na/K-pump function of ventricular myocytes. METHODS: We measured the [Na+]i dependence of the Na/K pump current, I(p). This relation was used as a calibration curve for [Na+]i based on I(p). We measured I(p) at the time of access and extrapolated [Na+] at the pump sites, i.e. subsarcolemmal [Na+], [Na+](subs), from the calibration curve. RESULTS: The extrapolated [Na+](subs) was significantly higher in CAVB (7.9 vs. 3.2 mM in control). The [Na+]i dependence of I(p) in CAVB myocytes was shifted to the right (range of [Na+](i): 0-20 mM). In resting cells, the I(p), i.e. steady state Na+ efflux, which matches Na+ influx, was higher in CAVB (0.25+/-0.02 vs. 0.47+/-0.06 pA/pF, P<0.05). Maximal I(p) density was not different, and DHO sensitivity was not altered. CONCLUSIONS: Hypertrophy in CAVB cells is associated with increased [Na+](subs). This results from an increase in Na+ influx, and a decreased sensitivity of I(p) for Na+ in the range of [Na+]i studied. There is no evidence for a decrease in total pump capacity or for a functional Na/K-ATPase isoform shift. The rise in Na+ contributes to the contractile adaptation and preservation of sarcoplasmic reticulum Ca2+ content at the low heart rates of the dog with CAVB. PMID- 12650882 TI - Cardiac ischemia causes inhibition of the Na/K ATPase by a labile cytosolic compound whose production is linked to oxidant stress. AB - OBJECTIVE: Intracellular Na rises rapidly during cardiac ischemia and this has been attributed to the combination of increased influx of Na via sodium-proton exchange and decreased activity of the Na/K ATPase. The aim of these studies was to investigate the effects of ischemia on Na/K ATPase function in Langendorff perfused rat hearts. METHODS: Na/K ATPase activity was determined by measuring ouabain-sensitive phosphate generation from ATP by cardiac homogenates. RESULTS: Global ischemia (15 and 30 min) caused a substantial reduction in Na/K ATPase function despite high substrate availability in the assay. When sarcolemmal membranes were purified away from the cytosol a profound activation of the Na/K ATPase was revealed following ischemia, indicating that the inhibition was due to the cytosolic accumulation of an inhibitor of Na/K ATPase. The half-life of the inhibitor in cardiac homogenates was 10+/-3 min at room temperature. Perfusion with the antioxidant MPG (1 mmol/l) reduced the accumulation of this inhibitor, however MPG was without effect on Na/K ATPase function when added directly to the Na/K ATPase activity assay. While the inhibitor reduced the activity of cardiac and brain forms of the Na/K ATPase in bioassay experiments, no effect was observed on the renal and skeletal muscle forms of the enzyme. CONCLUSIONS: An unstable cardiac and brain-specific inhibitor of the Na/K ATPase whose production is linked to oxidant stress, accumulates intracellularly during ischemia. Intracellular Na is a primary determinant of electro-mechanical recovery on reperfusion, so inhibition of the Na/K ATPase by this compound may be crucial in determining recovery from ischemia. PMID- 12650883 TI - Stretch-dependent slow force response in isolated rabbit myocardium is Na+ dependent. AB - OBJECTIVE: Stretch induces functional and trophic effects in mammalian myocardium via various signal transduction pathways. We tested stretch signal transduction on immediate and slow force response (SFR) in rabbit myocardium. METHODS: Experiments were performed in isolated right ventricular muscles from adult rabbit hearts (37 degrees C, 1 Hz stimulation rate, bicarbonate-buffer). Muscles were rapidly stretched from 88% of optimal length (L88) to near optimal length (L98) for functional analysis. The resulting immediate and slow increases in twitch force (first phase and SFR, respectively) were assessed at reduced [Na+]o or without and with blockade of stretch activated ion channels (SACs), angiotensin-II (AT1) receptors, endothelin-A (ET(A)) receptors, Na+/H+-exchange (NHE1), reverse mode Na+/Ca2+-exchange (NCX), or Na+/K+-ATPase. The effects of stretch on sarcoplasmic reticulum Ca2+-load were characterized using rapid cooling contractures (RCCs). Intracellular pH was measured in BCECF-AM loaded muscles, and action potential duration (APD) was assessed using floating electrodes. RESULTS: On average, force increased to 216+/-8% of the pre-stretch value during the immediate phase, followed by a further increase to 273+/-10% during the SFR (n=81). RCCs significantly increased during SFR, whereas pH and APD did not change. Neither inhibition of SACs, AT1, or ET(A) receptors affected the stretch-dependent immediate phase nor SFR. In contrast, SFR was reduced by NHE inhibition and almost completely abolished by reduced [Na+]o or inhibition of reverse-mode NCX, whereas increased SFR was seen after raising [Na+]i by Na+/K+ ATPase inhibition. CONCLUSIONS: The data demonstrate the existence of a delayed, Na+- and Ca2+-dependent but pH and APD independent SFR to stretch in rabbit myocardium. This inotropic response appears to be independent of autocrine/paracrine AT1 or ET(A) receptor activation, but mediated through stretch-induced activation of NHE and reverse mode NCX. PMID- 12650884 TI - What happens when cardiac Na channel function is compromised? 2. Numerical studies of the vulnerable period in tissue altered by drugs. AB - OBJECTIVE: The fate of an impulse arising from stimulation is determined by the ability of the wave front to recruit sufficient Na channels from adjacent cells. Previous numerical studies of mutant Na channels revealed both the velocity of a conditioning wave and the recruiting capacity of the front as determinants of the vulnerable period (VP), an interval within which excitation results in unidirectional conduction. Drugs that block excitatory Na channels in a voltage dependent manner, such as antiarrhythmics, abused substances and antidepressants, slow the restoration of Na conductance trailing an action potential and are associated with proarrhythmia and sudden cardiac death. We hypothesized that drug induced slowing of Na conductance recovery would flatten the Na conductance restoration gradient thereby reducing the recruiting capacity of a front, extending the VP and increasing the probability of unidirectional propagation. METHODS: In a cable of ventricular cells, we explored the sensitivity of the VP to voltage-dependent blockade. While varying the unbinding time constant from 100 ms to 5 s, we measured the Na conductance restoration gradient, the liminal length, the refractory period (RP) and the VP. RESULTS: Reducing the rate of drug unbinding flattened the restoration gradient, diminished the recruiting capacity of a premature impulse and extended the liminal length, RP and the VP. The VP was linearly dependent on the drug unbinding time constant. Rapidly unbinding drugs (time constant <1 s) reduced the liminal length below that of a quiescent cable. CONCLUSIONS: Slowing the unbinding rate of voltage-dependent drug block of Na channels extended the RP and the VP. Drugs with unbinding time constants greater than 1 s dramatically increased the probability of unidirectional propagation, reflecting increases in both the RP and the VP. This study provides a new mechanism linking Na channel function, compromised by voltage-dependent Na channel drug block, with proarrhythmic conditions that can lead to sudden cardiac death following premature stimulation. PMID- 12650885 TI - A novel LQT3 mutation implicates the human cardiac sodium channel domain IVS6 in inactivation kinetics. AB - The Long QT3 syndrome is associated with mutations in the cardiac sodium channel gene SCN5A. OBJECTIVE: The aim of the present study was the identification and functional characterization of a mutation in a family with the long QT3 syndrome. METHODS: The human cardiac sodium channel gene SCN5A was screened for mutations by single-stranded conformation polymorphism. The functional consequences of mutant sodium channels were characterized after expressing mutant and wild-type cRNAs in Xenopus oocytes by two-electrode voltage clamp measurements. RESULTS: SCN5A screening revealed an A-->G substitution at codon 1768, close to the C terminal end of domain IVS6, which changes an isoleucine to a valine. Functional expression of mutant I1768V-channels in Xenopus oocytes showed that the voltage dependence and slope factors of activation and inactivation were unchanged compared to wild-type channels. No difference in persistent TTX-sensitive current could be detected between wild-type and I1768V channels, a channel feature often increased in LQT3 mutants. However, I1768V mutant channels recovered faster from inactivation (2.4 times) than wild-type channels and displayed less slow inactivation. CONCLUSIONS: We postulate that severe destabilization of the inactivated state leads to increased arrhythmogenesis and QT prolongation in I1768V mutation carriers in the absence of a persistent inward sodium current. PMID- 12650886 TI - Serum and glucocorticoid inducible kinases in the regulation of the cardiac sodium channel SCN5A. AB - The serum and glucocorticoid inducible kinase SGK1 and its isoform SGK3 are both expressed in cardiac tissue. One of the functions of SGK1 is the phosphorylation and inactivation of the ubiquitin ligase Nedd4-2, which in turn could be shown to downregulate the voltage-gated Na+ channel SCN5A (hH1). The present study has been performed to test for a role of SGK1 and SGK3 in the regulation of SCN5A. To this end cRNA encoding the human Na+ channel SCN5A was injected into Xenopus laevis oocytes with or without cRNA encoding the wild-type kinases SGK1, the constitutively active kinase (S422D)SGK1, the inactive form K127NSGK1 or the wild type SGK3. SCN5A currents were activated by coexpression of either wild-type SGK1 or SGK3 or the constitutively active S422DSGK1. In contrast, the inactive mutant K127NSGK1 significantly decreased the currents. Moreover, coexpression of SGK3 significantly altered SCN5A gating, i.e. it hyperpolarized the activation threshold and depolarized the prepotential required for 50% availability of the channel. Opposite shifts of gating properties were elicited by mutation of serine to alanine (S483ASCN5A and S663ASCN5A) in the SGK consensus sequences of SCN5A. The present observations disclose a role of the kinases SGK1 and SGK3 in the regulation of cardiac Na+ channels. As SGK1 is upregulated by glucocorticoids, mineralocorticoids and a variety of inflammatory mediators and both kinases are activated by insulin and IGF1, the kinases could mediate effects of those hormones and mediators on cardiac function. PMID- 12650888 TI - [Anticoagulant clinic: a tool for reducing bleeding complications of oral anticoagulant treatment]. PMID- 12650887 TI - Effect of pacing and mexiletine on dispersion of repolarisation and arrhythmias in DeltaKPQ SCN5A (long QT3) mice. AB - OBJECTIVE: It has been suggested that both pacing and treatment with mexiletine may reduce torsade de pointes (TdP) arrhythmias in patients with long QT syndrome 3 (LQT3), but it is not fully understood how these interventions could prevent TdP. We therefore studied the effects of pacing and mexiletine in mice with a heterozygous knock-in DeltaKPQ SCN5A(Delta/+) deletion (SCN5A-Tg), a murine LQT3 model. METHODS: Three right and left ventricular monophasic action potentials (MAPs) were simultaneously recorded in Langendorff-perfused hearts of SCN5A-Tg and wild type (WT) littermates. AV block was induced, and pacing was performed at baseline and during mexiletine infusion (4 microg/ml). MAP recordings were analysed for action potential duration (APD), APD dispersion, and early afterdepolarisations (EADs) and related to spontaneous arrhythmias. RESULTS: After inducing AV block, SCN5A-Tg hearts were bradycardic [SCN5A-Tg 532+/-60 vs. WT 284+/-48 ms cycle length (CL, mean+/-S.E.M., P<0.05(*))]. EADs occurred in 16/18, and polymorphic ventricular tachycardia (pVT) in 11/18 SCN5A-Tg but not in 19 WT. SCN5A-Tg had longer APD than WT hearts*. At CL of 200 ms and longer, APD dispersion was higher in SCN5A-Tg [dispersion (APD70): 12+/-3 ms vs. 5+/-2 ms at CL=200 ms*], and increased to 35+/-4 ms* directly prior to pVT episodes. Sudden rate accelerations initially increased APD dispersion due to EADs and APD alternans in SCN5A-Tg, but pacing then reduced APD dispersion. Pacing suppressed (n=9/9) and prevented (n=49/50) pVT. Mexiletine shortened APD at long CL*, and suppressed pVT (n=4/5*), but did not prevent pVT during normal rhythm. CONCLUSIONS: Bradycardia, increased dispersion of APD and EADs provoke ventricular ectopy and pVT in SCN5A-Tg hearts. Ventricular pacing reduces APD dispersion, suppresses EADs and prevents pVT in SCN5A-Tg hearts. These effects provide a pathophysiological rationale for pacing in LQT3. PMID- 12650889 TI - [Central nervous tuberculosis in patients non-VIH: seven case reports]. AB - PURPOSE: Tuberculosis involving the central nervous system (CNS) is rarely observed in non immuno-compromised hosts. We report herin the various clinical, biological and radiological manifestations observed in 7 patients with CNS tuberculosis. METHODS: Clinical and biological records of 7 patients with CNS tuberculosis were retrospectively studied. All patients had encephalic CT-scan and MRI in the course of the disease. RESULTS: 5 women and 2 men with a mean age of 38.4 years initially initially presented with headache (n = 6), fever (n = 5), meningeal irritation (n = 3), localizing neurological signs (n = 1). Lumbar punction revealed lymphocytic meningitis (n = 6/7). Mycobacterium tuberculosis or bovis was isolated in 3 patients only. Cerebral tomodensitography or magnetic resonance imaging were initially normal in most of cases (n = 4/7), but discovered in the course of disease basilar meningitis (n = 6), hydrocephalus (n = 6), abcess or tuberculoma (n = 4). In all the patients, initiation of the treatment was complicated by clinical and/or biological deterioration, called paradoxal reaction, leading in all cases to glucocorticoid adjunction, with various final results. Indeed, 4 patients developed neurological sequelae. No patient died. CONCLUSION: CNS tuberculosis is a rare disease in non immunocompromised patients whose diagnostic may be difficult due to the absence of specific clinical symptoms, negative initial radiological examination, as well as delayed and often negative bacterial isolation. Paradoxal reaction appeared to be frequent despite specific antibiotherapy and underlines the beneficial effects of addictive corticosteroids. PMID- 12650890 TI - [Neuroradiologic manifestations of central nervous system tuberculosis in 122 adults]. AB - PURPOSE: Central nervous system (CNS) tuberculosis remains a public health problem, particularly in developing countries. The aim of this study is to characterize neuroradiologic findings of various intracranial lesions. METHODS: We retrospectively reviewed data of 122 patients with CNS tuberculosis, without immunosuppression. CT scan was performed in all patients, whereas 17 patients had CT scan and MRI. RESULTS: We included 74 women (61%) and 48 men (39%) with a mean age of 37 years (17 -88y). 18 patients (14,7%) had a history of tuberculosis. Tuberculous meningitis was the most frequent clinical presentation (119 cases). Mycobacterium tuberculosis was isolated in cerebrospinal fluid of 18 patients (15%). Several types of lesions were identified : hydrocephalus (35 cases), tuberculomas (29 cases), leptomeningitis (26 cases), infarction (15 cases), abcesses (2 cases). Hydrocephalus was associated to other lesions in 26 cases. Communication hydrocephalus was present in 28 cases. Multiple tuberculomas were seen in 23 cases (80%), with miliary aspects in some cases. In 3 cases, tuberculoma was present without meningitis. Patients with leptomeningitis showed thick meningeal contrast enhancement involving all basal cisterns. Infarction resulted from arterial englobement or embols, and involved the area of middle cerebral artery (12 cases). CONCLUSION: Central nervous system tuberculosis has different appearences, mostly hydrocephalus and tuberculomas. MR with contrast is necessary for diagnosis and for follow-up during treatment. PMID- 12650891 TI - [Amoebic liver abscess. Study of 20 cases with literature review]. AB - PURPOSE: The management of amoebic liver abscess includes antiamoebic drugs combined or not with percutaneous puncture or surgical drainage. This study was to suggest a decision tree for the therapeutic approach of such feature. METHODS: We report a retrospective analysis of 20 imported cases with amoebic liver abscesses admitted at the Department of Tropical Diseases during 1995-1999 at the Bordeaux University Hospital Centre, France, and a review of the literature. RESULTS: The twenty patients were 14 males and 6 females, mainly 20 to 40 years old. The clinical presentation was mainly accounting a painful liver enlargement with hyperthermia. The echographic picture was mostly represented by a unique liver element located at the liver right lobe. They were numerous in an HIV infected patient. Thirteen patients have been treated using a medical therapeutic approach. A percutaneous puncture has been necessary for 4 cases. A percutaneous drainage has been realised for two patients as regard to the persistence of the hepatalgia occurrence. A surgical drainage has been experienced by two patients after a lack of efficacy of a percutaneous drainage, after rupture of an abscess treated medically, respectively. A review of the literature and the analysis of the 20 cases history have been used to determine a therapeutic algorithm. CONCLUSION: The occurrence of immediate complications at onset must indicate a first line surgical drainage procedure. Beside this situation, risk factors for rupture must be assessed (high size abscess, pejorative localization), as well as poor prognosis feature (liver failure, bacteraemia). If no pejorative condition occurs, a first-line exclusive medical approach can be undertaken with a clinical efficacy evaluation at H72. Otherwise, the indication of the percutaneous drainage must be discussed. PMID- 12650892 TI - [Oral anticoagulants in the elderly]. AB - PURPOSE: Atrial fibrillation and venous thromboembolism are particularly frequent in the elderly. Whether or not prescribe oral anticoagulant treatment in the elderly is therefore a common question for the physician. Despite the benefits of anticoagulation demonstrated in clinical trials, oral anticoagulant therapy is underused in the elderly. CURRENT KNOWLEDGE AND KEY POINTS: Indications for oral anticoagulation are discussed specifically in the elderly with a literature review. Only the length of anticoagulation treatment after a venous thromboembolism remained a purpose of discussion regarding the severity of the pathology. The frequency of systemic thromboembolism in nonvalvular atrial fibrillation is increasing with age. Oral anticoagulation reduces this risk. This benefice is to compare with the increasing rate of major bleeding complications in the elderly and in patient who had stroke, hyper-tension, diabetes mellitus or gastrointestinal bleeding. The objective of this article is to focus on the mode to measure oral anticoagulant benefice/risk ratio in the elderly and to propose several ways to minimize the risk for bleeding. FUTURE PROSPECTS AND PROJECTS: The potential drug side effect severity of oral anticoagulation must lead to find the "reasonable" clinical indications in term of benefice/risk ratio and what measures should be take to increase the safety of oral anticoagulation in the elderly. The comprehensive geriatric evaluation should be considered as a decision-aid tool in long-term oral anticoagulation in the frail elderly. Anticoagulation clinics, informatics'-prescription coupled with dose-adaptation decision-aid adapted to the elderly should be helpful in this research of quality. Finally, prescribers education supports must insist on the early course of therapy that is at higher risk of bleeding. PMID- 12650893 TI - [Severe Moorens ulcer: efficacy of monthly cyclophosphamide intravenous pulse treatment]. AB - PURPOSE: Mooren's ulcer (MU) is a chronic peripheral corneal ulceration featuring conjunctival immunoglobulin deposits. It is considered as the result of a limbic immune process with hyperactivation of T and B lymphocytes. The etiology remains unknown. The response to topical steroid therapy and surgical procedures usually poor and the visual outcome can be devastating. METHODS: Clinical follow-up of 3 patients who had rebel MU to conventional therapy, and were treated with 1g monthly intravenous cyclophosphamide. RESULTS: First patient was a 24-years-old man who had MU in his left eye. The response to surgical procedure and intravenous steroid treatment was poor and corneal perforation occurred. The affected cornea healed after 9 months of Cy treatment. The second patient was a 50-years-old man who had MU in his left eye, which did not improved with lamellar keratoplasty and topical steroid therapy. Corneal healing was obtained after 20 months of Cy treatment. The third patient was a 70-years-old man who presented with a furrowed MU in his right eye which healed with conjunctival resection and 4 months of Cy perfusion. No adverse effects of Cy was noted as opposed to Cy given orally. CONCLUSION: We report the effectiveness of 1g monthly intravenous cyclophosphamide (Cy) treatment in rebel MU. We suggest that immunosuppressive therapy using IV monthly Cy may be proposed in severe rebel MU. PMID- 12650894 TI - [Inhibitors of TNFalpha]. AB - INTRODUCTION: TNFalpha inhibitors etanercept and infliximab, are a new and very exciting drugs. Etanercept is administered subcutaneously, and infliximab by intravenous perfusions. EXEGESIS: They are indicated in severely rheumatoid arthritis resistant to treatment with methotrexate, and in Crohn's disease refractory to immunosuppressive agents. Their safety profile is good, but the risk of infections is increased. CONCLUSION: Their indication in other inflammatory diseases need further studies. PMID- 12650895 TI - [Cannabis arteritis: four new cases]. AB - INTRODUCTION: The responsibility of cannabis in juvenile thromboangeitis has been suggested for few years. We describe four new cases. EXEGESIS: Young men presented with distal arteriopathy of the lower limbs in 3 cases, and of the left upper limb in the remaining patient. Symptoms occurred progressively, distal pulses had disappeared, and distal necrosis was constant. Three patients suffered from Raynaud phenomenon, none of them presented with venous thrombosis. Radiologic evaluation revealed distal abnormalities in all cases, and proximal arterial thrombosis in one case. The four patients were cannabis smokers for at least four years. With cannabis interruption and symptomatic treatment, lesions improved for three patients. For one of them, recurrence of arteriopathy occurred when he resumed to smoke cannabis. For the fourth one who never stopped cannabis, an amputation was necessary. CONCLUSION: Search for cannabis use is important because interruption may improve prognosis. PMID- 12650896 TI - [Disappearance of auto-antibody-induced haemolysis after resection of a gastric stromal tumor. Case report and review of the literature]. AB - INTRODUCTION: There are two types of autoimmune haemolytic anemia, idiopathic or associated to other disorders (infections, thyroidal diseases, cancer.). The treatment of this particular anemia depends on the primary or secondary feature. EXEGESIS: We report the case of a 76-year old woman, presenting an autoimmune haemolytic anemia with a positive direct Coombs test in IgG. The corticodependance of this anemia after 6 months of treatment had indicated a splenectomy. During surgery, a gastric stromal tumor was discovered. After removal of the tumor and splenectomy, the haematological symptomatology quickely disappeared and induced the negativation of the Coombs test within a post operative period going from 7 to 14 months, without any immunosuppressor treatment. CONCLUSION: Splenectomy does not induce the negativation of a Coombs test in an autoimmune haemolytic anemia, someaning that there is a link between the anemia and the tumor. This observation is completed by a review the literature. PMID- 12650897 TI - [Atypical tophaceous gout]. PMID- 12650898 TI - [Sjogren's syndrome associated with multiple myeloma]. PMID- 12650899 TI - [Juvenile temporal arteritis: a benign disease]. PMID- 12650900 TI - Acridine mutagenesis of zebrafish (Danio rerio). AB - Mutagenesis screening, in which heritable traits are isolated following damage to the genome, is a powerful approach for investigating gene function. Among vertebrate model organisms, the zebrafish (Danio rerio) is ideally suited to mutagenesis screens. The success of large-scale screens is dependent on the way in which changes are identified. The type of damage induced is also pivotal. Single base coding region deletions and insertions are suited to abolition of gene function whilst inducing a small physical alteration to the genome. Such mutations are not commonly found following mutagenesis schemes reported to date. Here, we show that an acridine mutagen, ICR191, which in other model organisms frequently induces single base deletions and insertions, is mutagenic in zebrafish. ICR191 induces hallmark phenotypes associated with genetic damage in treated embryos. Alterations are heritable. Offspring of mutagenised fish had mutations in a marker gene and were found to produce offspring with abnormal development. Using an adaptation of a molecular mutation detection method, fluorescent arbitrary primed PCR, we identified an induced alteration directly. The estimated frequency of induced mutations was sufficiently high to make it feasible to employ this approach for mutagenesis screening. PMID- 12650901 TI - Single nucleotide polymorphism--disease relationships: statistical issues for the performance of association studies. AB - There is increasing concern about the low reproducibility of findings of genetic association studies in complex diseases. In the present paper, we argue that it might be beneficial to learn from epidemiology and the methodology for clinical trials which address problems also crucial for genetic association studies. We propose a two-step sequential approach in order to (a) allow a study--internal control of reproducibility and (b) stop a study or lab analysis prematurely to save valuable specimen, time and costs. Additionally, minimal information is proposed which should be presented in publications about association studies in order to facilitate reproducibility studies by other authors. PMID- 12650902 TI - Changes in the human mitochondrial genome after treatment of malignant disease. AB - Mitochondrial DNA (mtDNA) is the only extrachromosomal DNA in human cells. The mitochondrial genome encodes essential information for the synthesis of the mitochondrial respiratory chain. Inherited defects of this genome are an important cause of human disease. In addition, the mitochondrial genome seems to be particularly prone to DNA damage and acquired mutations may have a role in ageing, cancer and neurodegeneration. We wished to determine if radiotherapy and chemotherapy used in the treatment of cancer could induce changes in the mitochondrial genome. Such changes would be an important genetic marker of DNA damage and may explain some of the adverse effects of treatment. We studied samples from patients who had received radiotherapy and chemotherapy for point mutations within the mtDNA control region, and for large-scale deletions. In blood samples from patients, we found a significantly increased number of point mutations compared to the control subjects. In muscle biopsies from 7 of 8 patients whom had received whole body irradiation as well as chemotherapy, the level of a specific mtDNA deletion was significantly greater than in control subjects. Our studies have shown that in patients who have been treated for cancer there is an increased level of mtDNA damage. PMID- 12650903 TI - High gradient magnetic cell sorting and internal standardisation substantially improve the assay for somatic mutations at the glycophorin A (GPA) locus. AB - In the MACS-BR6 version of the GPA assay [Int. J. Radiat. Res. 70 (1996) 131] variant red blood cells (RBC) are isolated from 5 x 10(8) normal RBC by magnetic cell separation (MACS) before detection and quantification by immunolabelling and flow cytometry as in the classical BR6 assay. In the present work it is described how the MACS-BR6 assay is improved by internal standardisation with FITC-labelled RBC. This modification of the assay has the advantage that (i) the analysis of variants is not disturbed by the overwhelming number of normal RBC that (ii) the precision of the assay is improved and finally that (iii) a sufficient number of variants is available for further investigations. Tn positive RBC behave in MACS like variants. It is demonstrated that in normal individuals Tn cells (frequency: approximately 2 x 10(-8)) do not disturb the assay. PMID- 12650904 TI - Biological activities of organic compounds adsorbed onto ambient air particles: comparison between the cities of Teplice and Prague during the summer and winter seasons 2000-2001. AB - The capital of the Czech Republic, Prague, appears today to be one of the most polluted residential areas in the country, whereas air pollution in the Northern Bohemia region (the former "Black Triangle Region") has substantially decreased during the last decade, especially with respect to the gaseous pollutant SO(2). This study evaluated the biological activities of complex mixtures of organic compounds adsorbed onto ambient air particles (PM10) collected during the summer and winter seasons of 2000-2001 at three monitoring sites--Teplice (TP), Prague Smichov (PRG-SM) (city centre) and Prague-Libus (PRG-LB) (suburban area). The following short-term in vitro assays with strikingly different endpoints were used: a bacterial mutagenicity test using the Salmonella typhimurium tester strain TA98 and YG1041, an acellular assay (CT DNA) combined with 32P postlabelling to evaluate DNA adduct-forming potency and the chick embryotoxicity screening test (CHEST). The results of the mutagenicity test with the YG1041 strain, the acellular genotoxicity (DNA adducts) and the embryotoxicity tests responded to the amount of eight carcinogenic polycyclic aromatic hydrocarbons (PAHs) analysed in the EOM (dichloromethane extractable organic matter) samples tested. Nevertheless, the biological effects of the EOM did not differ between locations. The highest biological activity of the ambient air in terms of organic compounds associated with particles (per unit volume of air) was seen in the Prague city centre during both summer and winter seasons. At this location, B[a]P concentration ranged from 0.1 to 8.9 ng/m(3) (mean 0.3 and 3.6 ng/m(3) for summer and winter seasons, respectively), 13 PAHs ranged from 11 to 343 ng/m(3) (mean 52 and 160 ng/m(3) for summer and winter seasons, respectively). Generally, using in vitro tests, higher ambient air activity was found in the winter season as compared with the summer season at all three monitoring sites (TA98 +S9, approximately 4-fold; YG1041 -S9, approximately 5-fold; YG1041 +S9, approximately 8-fold; CT DNA +S9, approximately 10-fold; CHEST, approximately 10-fold; B[a]P, carcinogenic PAHs and total PAHs analysed, more than 10-fold). The different proportions of individual PAHs found in the summer and winter samples suggested traffic as a major emission source in the summer and, additionally, residential heating in the winter season at all three monitoring sites. The DNA adduct patterns resulting from the in vitro acellular assay also demonstrated similar major emission sources at all three locations. The study shows that particle bound carcinogenic-PAH concentrations may be taken as an index for the biologically active (mutagenic, genotoxic, embryotoxic) components in air particulate samples. Therefore, high-quality monitoring data of carcinogenic PAHs may be useful for epidemiological studies of the impact of air pollution on the health of the population and for helping decision makers to improve our environment. PMID- 12650905 TI - A single nucleotide deletion of 293delT in SEDL gene causing spondyloepiphyseal dysplasia tarda in a four-generation Chinese family. AB - Spondyloepiphyseal Dysplasia Tarda (SEDT; MIM 313400) is a rare genetically heterogeneous disorder of vertebral and epiphyseal growth resulting in disproportionally short-trunked short stature, barrel-shaped chest, and dysplasia of the large joints. It is caused by the mutations of SEDL gene. The distinctive radiological signs and the X-linked mode of inheritance make it easy to diagnose. Here a four-generation Chinese SEDT family has been analyzed and the disease causing mutation has been found. After polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) analysis and DNA sequencing, a previously unreported deletion of T in exon 5 of SEDL gene (i.e. 293delT) was observed and seven individuals in the family carried the mutation. It results in frameshift and a putative truncated protein with the 97 N-terminal amino acids, and 9 changed amino acids. Therefore, loss of function of the gene could be predicted. However, this mutation has not been detected in 50 age and sex matched unrelated controls. PMID- 12650906 TI - Characterization of trimethylpsoralen as a mutagen for mouse embryonic stem cells. AB - Given a large number of genes with unknown functions in model organisms, collections of mutants are valuable resources for studying gene function. For the mouse, embryonic stem cell technology offers the possibility to manipulate the genome and select for mutations in vitro. Mutant mice can then be generated from clones of interest to study the phenotype of these animals. We manipulate the genome of mouse embryonic stem (ES) cells chemically using the mutagen trimethylpsoralen (TMP). TMP predominantly causes deletions in the genome of Caenorhabditis elegans and Escherichia coli, but has not been established as a mutagen in mammalian systems yet. We have characterized TMP as a mutagen for mouse ES cells regarding death rates, mutation frequencies, and mutation spectrum. Allowing for 12.5% of cell survival, the mutation frequency at the mouse hypoxanthine-guanine phosphoribosyltransferase (Hprt) locus was 3.5 x 10( 5) on average. The characterization of a non-redundant set of 17 Hprt-deficient ES clones revealed that only 12% of clones contained genomic deletions and almost 50% were point mutations. Base substitutions were mostly transversions and all affected AT base pairs. We conclude that the mutation spectrum of TMP in mouse ES cells is different from that observed in C. elegans and E. coli. PMID- 12650907 TI - Mutation spectra of smoky coal combustion emissions in Salmonella reflect the TP53 and KRAS mutations in lung tumors from smoky coal-exposed individuals. AB - Nonsmoking women in Xuan Wei County, Yunnan Province, China who use smoky coal for cooking and heating in poorly ventilated homes have the highest lung cancer mortality rate in China, and their lung cancer is linked epidemiologically to their use of smoky coal. The emissions contain 81% organic matter, of which 43% is polycyclic aromatic hydrocarbons (PAHs). Exposure assessment and molecular analysis of the lung tumors from nonsmoking women who use smoky coal strongly indicate that PAHs in the emissions are a primary cause of the elevated lung cancer in this population. Here we have determined the mutation spectra of an extract of smoky coal emissions in Salmonella TA98 and TA100; the extract was not mutagenic in TA104. The extract was 8.7 x more mutagenic in TA100 with S9 than without (8.7 rev/microg versus 1.0 rev/microg) and was >3 x more mutagenic in TA100 than in TA98--consistent with a prominent role for PAHs in the mutagenicity of the extract because PAHs are generally more mutagenic in the base-substitution strain TA100 than in the frameshift strain TA98. The extract induced only a hotspot mutation in TA98; another combustion emission, cigarette smoke condensate (CSC), also induces this single class of mutation. In TA100, the mutation spectra of the extract were not significantly different in the presence or absence of S9 and were primarily (78-86%) GC --> TA transversions. This mutation is induced to a similar extent by CSC (78%) and the PAH benzo[a]pyrene (B[a]P) (77%). The frequency of GC --> TA transversions induced in Salmonella by the extract (78 86%) is similar to the frequency of this mutation in the TP53 (76%) and KRAS (86%) genes of lung tumors from nonsmoking women exposed to smoky coal emissions. The mutation spectra of the extract reflect the presence of PAHs in the mixture and support a role for PAHs in the induction of the mutations and tumors due to exposure to smoky coal emissions. PMID- 12650909 TI - Extremely complex pattern of microsatellite mutation in the germline of wheat exposed to the post-Chernobyl radioactive contamination. AB - The molecular structure of rare variants at 13 microsatellite loci found in a population of wheat plants grown for one generation in the heavily contaminated 30 km exclusion zone around the Chernobyl Nuclear Power Plant and in a control population was compared. Evidence for rare alterations (variants) was obtained for all 13 loci, including gain and loss of repeats, as well as the complete loss of microsatellite bands. The ratio between gains and losses among variants in the control group was similar to that in the exposed group. Sequencing of variants at six microsatellite loci found in the exposed population revealed extremely complex pattern of germline mutations, including complete deletions of loci, a bias towards mutations with gains and losses of multiple repeat units, and relatively frequent insertions of DNA of unknown origin. The occurrence of large deletions at two loci may be attributed to direct and inverted repeats sequences located just upstream and downstream of the array. The results of our study also suggest that the majority of mutations within the studied wheat microsatellite loci are represented by gains and losses of multiple repeat units, implying that a simple model of replication slippage cannot account for mutation events at these loci. Our data also support the conclusion that the spectra of spontaneous and radiation-induced mutation in wheat may be similar. PMID- 12650908 TI - Ionizing radiation-induced Rad51 nuclear focus formation is cell cycle-regulated and defective in both ATM(-/-) and c-Abl(-/-) cells. AB - In eukaryotes, DNA double-strand breaks (DSBs) can be repaired by either non homologous end-joining (NHEJ) or homologous recombination (HR) pathways. Rad50 protein is a component of the Rad50/NBS1/Mre11 nuclease complex that functions in both the NHEJ and recombinational repair of DNA DSBs. On the other hand, Rad51 protein, a homolog of bacterial RecA and a member of the Rad52 epistasis group, plays a crucial role exclusively in the recombinational repair pathway. We analyzed the effects of cell cycle progression and genetic background on the ionizing radiation (IR)-induced Rad51 and Rad50 repair focus formation. Herein, we demonstrated that IR-induced Rad51, but not Rad50, nuclear focus formation was cell cycle-dependent. Furthermore, IR-induced Rad51 focus formation was defective in AT and c-Abl(-/-) cells, but not wild type or NBS cells. A decreased and delayed formation of Rad51 foci-containing nuclei was observed in AT cells upon IR, whereas in c-Abl(-/-) cells a decreased but not delayed formation of Rad51 foci-containing nuclei was observed. In conclusion, effective and prompt IR induced Rad51 focus formation is cell cycle-regulated and requires both ATM and c Abl. PMID- 12650910 TI - Y chromosome instability in lymphoproliferative disorders. PMID- 12650911 TI - Mechanism of DNA strand breakage induced by photosensitized tetracycline-Cu(II) complex. AB - Tetracyclines (TCs) in combination with Cu(II) ions exhibited significant DNA damaging potential vis a vis tetracyclines per se. Interaction of tetracyclines with DNA resulted in alkylation at N-7 and N-3 positions of adenine and guanine bases, and caused destabilization of DNA secondary structure. Significant release of acid-soluble nucleotides from tetracycline-modified DNA upon incubation with S(1) nuclease ascertained the formation of single stranded regions in the DNA. Also, the treatment of tetracycline-modified DNA with 0.1 and 0.5M NaOH resulted in 62 and 76% hydrolysis compared to untreated control. Comparative alkaline hydrolysis of DNA modified with tetracycline derivatives showed differential DNA damaging ability in the order as DOTC > DMTC > TC > OTC > CTC. Addition of Cu(II) invariably augmented the extent of tetracycline-induced DNA damage. The alkaline unwinding assay clearly demonstrated the formation of approximately six strand breaks per unit DNA at 1:10 DNA nucleotide/TC molar ratio in the presence of 0.1mM Cu(II) ions. At a similar Cu(II) concentration, a progressive transformation of covalently closed circular (CCC) (form-I) plasmid pBR322 DNA to forms-II and -III was noticed with increasing tetracycline concentrations. The results obtained with the free-radical quenchers viz. mannitol, thiourea, sodium benzoate and superoxide dismutase (SOD) suggested the involvement of reactive oxygen species in the DNA strand breakage. It is concluded that the tetracycline Cu(II)-induced DNA damage occurs due to (i) significant binding of tetracycline and Cu(II) with DNA, (ii) methyl group transfer from tetracycline to the putative sites on nitrogenous bases, and (iii) metal ion catalyzed free-radical generation in close vicinity of DNA backbone upon tetracycline photosensitization. Albeit, the DNA alkylation and strand cleavage are repairable lesions, but any defect in the critical repair pathway may augment the damage accumulation and mutagenesis. PMID- 12650912 TI - Identification of a novel mutation in WFS1 in a family affected by low-frequency hearing impairment. AB - Previously we confirmed linkage of autosomal dominantly inherited low-frequency sensorineural hearing impairment (LFSNHI) in a German family to the genetic locus DFNA6/DFNA14 on chromosome 4p16.3 close to the markers D4S432 and D4S431. Analysis of data from the Human Genome Project, showed that WFS1 is located in this region. Mutations in WFS1 are known to be responsible for Wolfram syndrome (DIDMOAD, MIM #606201), which follows an autosomal recessive trait. Studies in low-frequency hearing loss families showed that mutations in WFS1 were responsible for the phenotype. In all affected family members analysed, we detected a missense mutation in WFS1 (K705N) and therefore confirm the finding that the majority of mutations responsible for LFSNHI are missense mutations which localise to the C-terminal domain of the protein. PMID- 12650913 TI - Comment on "The radiation-induced lesions which trigger the bystander effect" by J.F. Ward [Mutat. Res. 499 (2002) 151-154]. PMID- 12650915 TI - Strong support from the scientific community to the idea of establishing a European Research Council within the Life Sciences. PMID- 12650916 TI - A novel strategy to design binding molecules harnessing the modular nature of repeat proteins. AB - Repeat proteins, such as ankyrin or leucine-rich repeat proteins, are ubiquitous binding molecules, which occur, unlike antibodies, intra- and extracellularly. Their unique modular architecture features repeating structural units (repeats), which stack together to form elongated repeat domains displaying variable and modular target-binding surfaces. Based on this modularity, we developed a novel strategy to generate combinatorial libraries of polypeptides with highly diversified binding specificities. This strategy includes the consensus design of self-compatible repeats displaying variable surface residues and their random assembly into repeat domains. We envision that such repeat protein libraries will be highly valuable sources for novel binding molecules especially suitable for intracellular applications. PMID- 12650917 TI - BTK-2, a new inhibitor of the Kv1.1 potassium channel purified from Indian scorpion Buthus tamulus. AB - A novel inhibitor of voltage-gated potassium channel was isolated and purified to homogeneity from the venom of the red scorpion Buthus tamulus. The primary sequence of this toxin, named BTK-2, as determined by peptide sequencing shows that it has 32 amino acid residues with six conserved cysteines. The molecular weight of the toxin was found to be 3452 Da. It was found to block the human potassium channel hKv1.1 (IC(50)=4.6 microM). BTK-2 shows 40-70% sequence similarity to the family of the short-chain toxins that specifically block potassium channels. Multiple sequence alignment helps to categorize the toxin in the ninth subfamily of the K+ channel blockers. The modeled structure of BTK-2 shows an alpha/beta scaffold similar to those of the other short scorpion toxins. Comparative analysis of the structure with those of the other toxins helps to identify the possible structure-function relationship that leads to the difference in the specificity of BTK-2 from that of the other scorpion toxins. The toxin can also be used to study the assembly of the hKv1.1 channel. PMID- 12650918 TI - Structural study of a single-point mutant of Sulfolobus solfataricus alcohol dehydrogenase with enhanced activity. AB - Alcohol dehydrogenase from Sulfolobus solfataricus (SsADH) is the only enzyme from Archaea among the structurally studied members of the medium-chain ADH family described so far. Here, we present the three-dimensional structure of the apo form of the mutant N249Y which exhibits increased catalytic activity when compared to the wild-type enzyme. The substitution, located in the coenzyme binding domain, decreases the affinity for NAD(H) cofactor. The rearrangement of segments 248-250 and 270-275, induced by the mutation, suggests an explanation for the lower coenzyme affinity. This study also highlights the role in SsADH catalysis of the flexible loops located at the interface between the catalytic and the coenzyme domains. PMID- 12650920 TI - Selection based on the folding properties of proteins with ribosome display. AB - Ribosome display is a powerful tool for selecting and evolving protein functions through ligand-binding. Here, this in vitro system was used to perform selection based on the folding properties of proteins, independent of specific ligand binding. The selection is based on two properties of misfolded proteins: (1) increased sensitivity to proteolysis and (2) greater exposure of hydrophobic area. By targeting these properties, we show that compactly folded and soluble proteins can be enriched over insoluble and random coil proteins. This approach may be especially useful for selection and evolution of folded proteins from random sequence libraries. PMID- 12650919 TI - Effects of interactions of hepatocyte nuclear factor 4alpha isoforms with coactivators and corepressors are promoter-specific. AB - The gene encoding hepatocyte nuclear factor 4alpha (HNF4alpha) possesses two alternative promoters responsible for developmental and tissue-specific expression of HNF4alpha1 and HNF4alpha7. The two isoforms possess different N termini and exhibit distinct transactivation properties. We show here for the first time that the effects mediated by HNF4alpha isoforms in concert with three different coregulators result in promoter-specific responses. Transcript levels of silencing mediator for retinoid and thyroid receptors and glucocorticoid receptor interacting protein-1 in the liver are reduced at birth, a time point when many genes are strongly activated, suggesting that the effects of coregulators on HNF4alpha activity in vivo could be determined by the levels of their expression as well as by the target promoter. PMID- 12650921 TI - Activation mechanism of c-Jun amino-terminal kinase in the course of endodermal differentiation of P19 embryonic carcinoma cells. AB - c-Jun amino-terminal kinase (JNK) is known to be activated and play critical roles during neural and endodermal differentiation of P19 embryonic carcinoma cells. In this study we demonstrated that of the two upstream protein kinases of JNK, only MKK4 activity was substantially enhanced in the endodermally differentiating P19 cells. This enhanced activity of MKK4 stemmed from the increased expression of MKK4 and its activation by phosphorylation. Activated MKK4 and JNK were localized in both nucleus and cytoplasm of the differentiating cells, while they were localized only in the nucleus in the undifferentiated cells, suggesting multiple roles of JNK in the course of the endodermal differentiation of P19 cells. PMID- 12650922 TI - Discodermolide interferes with the binding of tau protein to microtubules. AB - We investigated whether discodermolide, a novel antimitotic agent, affects the binding to microtubules of tau protein repeat motifs. Like taxol, the new drug reduces the proportion of tau that pellets with microtubules. Despite their differing structures, discodermolide, taxol and tau repeats all bind to a site on beta-tubulin that lies within the microtubule lumen and is crucial in controlling microtubule assembly. Low concentrations of tau still bind strongly to the outer surfaces of preformed microtubules when the acidic C-terminal regions of at least six tubulin dimers are available for interaction with each tau molecule; otherwise binding is very weak. PMID- 12650923 TI - C-terminal 15 kDa fragment of cytoskeletal actin is posttranslationally N myristoylated upon caspase-mediated cleavage and targeted to mitochondria. AB - To detect the posttranslational N-myristoylation of caspase substrates, the susceptibility of the newly exposed N-terminus of known caspase substrates to protein N-myristoylation was evaluated by in vivo metabolic labeling with [(3)H]myristic acid in transfected cells using a fusion protein in which the query sequence was fused to a model protein. As a result, it was found that the N terminal nine residues of the newly exposed N-terminus of the caspase-cleavage product of cytoskeletal actin efficiently direct the protein N-myristoylation. Metabolic labeling of COS-1 cells transiently transfected with cDNA coding for full-length truncated actin (tActin) revealed the efficient incorporation of [(3)H]myristic acid into this molecule. When COS-1 cells transiently transfected with cDNA coding for full-length actin were treated with staurosporine, an apoptosis-inducing agent, an N-myristoylated tActin was generated. Immunofluorescence staining coupled with MitoTracker or fluorescence tagged phalloidin staining revealed that exogenously expressed tActin colocalized with mitochondria without affecting cellular and actin morphology. Taken together, these results demonstrate that the C-terminal 15 kDa fragment of cytoskeletal actin is posttranslationally N-myristoylated upon caspase-mediated cleavage during apoptosis and targeted to mitochondria. PMID- 12650924 TI - Crosslinking of CD44 on human osteoblastic cells upregulates ICAM-1 and VCAM-1. AB - Cell adhesion of osteoblasts and osteoclastic precursors of hematopoietic origin is a prerequisite for osteoclast maturation. We have investigated the relevance of osteoblast-matrix binding and regulation of adhesion molecules to this process. Human osteoblastic cells highly expressed CD44, a major receptor for hyaluronan present in the surrounding bone matrix. Crosslinking of CD44 on osteoblastic cells with specific antibodies augmented the expression of intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1. Hyaluronan, the major ligand of CD44, also up-regulated ICAM-1 expression. Stimulation of CD44 on osteoblastic cells amplified their adhesion to monocytic cells through ICAM-1 and VCAM-1. These results suggest that such crosstalk among distinct adhesion molecules may be relevant to bone metabolism, including osteoclastogenesis. PMID- 12650925 TI - FBPase is in the nuclei of cardiomyocytes. AB - Intracellular localization of FBPase in the cardiac muscle of the pig was studied by immunohistochemistry. In contrast to data from skeletal muscle [Gizak, A., Rakus, D. and Dzugaj, A. (2003) Histol. Histopathol. 18, 135-142], in cardiomyocytes FBPase was present not only in the cytoplasm, but surprisingly, also in the nucleus. Results of the microscopic investigation were confirmed by immunoblotting, measurement of FBPase activity in isolated cardiomyocyte nuclei and by determination of the nuclear FBPase I(0.5) toward adenosine monophosphate (AMP), which was the same as for the purified enzyme. PMID- 12650926 TI - Sphingosine-1-phosphate inhibits acid sphingomyelinase and blocks apoptosis in macrophages. AB - Sphingosine-1-phosphate (Sph-1-P) regulates critical cellular functions including cell proliferation, differentiation, angiogenesis, and cell survival. However, its mechanisms of action are incompletely understood. Here, we show a novel biological effect of Sph-1-P: inhibition of acidic sphingomyelinase (A-SMase) activity in apoptotic bone marrow-derived macrophages. A-SMase catalyzes the conversion of sphingomyelin to ceramides, which are pro-apoptotic. This action of Sph-1-P prevents the accumulation of ceramides and blocks apoptosis, thereby promoting survival of the macrophages. PMID- 12650927 TI - The Escherichia coli twin-arginine translocase: conserved residues of TatA and TatB family components involved in protein transport. AB - The Escherichia coli Tat system serves to export folded proteins harbouring an N terminal twin-arginine signal peptide across the cytoplasmic membrane. In this report we have studied the functions of conserved residues within the structurally related TatA and TatB proteins. Our results demonstrate that there are two regions within each protein of high sequence conservation that are critical for efficient Tat translocase function. The first region is the interdomain hinge between the transmembrane and the amphipathic alpha-helices of TatA and TatB proteins. The second region is within the amphipathic helices of TatA and TatB. In particular an invariant phenylalanine residue within TatA proteins is essential for activity, whereas a string of glutamic acid residues on the same face of the amphipathic helix of TatB is important for function. PMID- 12650928 TI - Cu,Zn-superoxide dismutase of Saccharomyces cerevisiae is required for resistance to hyperosmosis. AB - Here we analyzed the role of the antioxidant response in Saccharomyces cerevisiae adaptation to hyperosmotic stress. We show that Cu,Zn-superoxide dismutase (SOD1) plays a fundamental role in this adaptation process since under hyperosmosis SOD1 mutants lead to high protein oxidation levels and show a sensitive phenotype, which is reversed by the addition of N-acetylcysteine to the medium. Pretreatment with MnCl(2), a superoxide scavenger, improves the survival of the sod1 strain upon hyperosmosis. Additionally, we show that upon hyperosmotic shock there is a small and transient increase in SOD1 transcript levels, regulated by the protein kinase A-cAMP and SKN7 pathways. PMID- 12650929 TI - Human T-cell leukemia virus type 1 Tax protein stimulates the interferon responsive enhancer element via NF-kappaB activity. AB - Gene expression in Rat-1 fibroblast cells transformed by Tax from human T-cell leukemia virus type 1 was studied using the reverse transcriptase polymerase chain reaction differential display technique. The analysis revealed eight genes that were upregulated and one gene that was suppressed in Tax-transformed cells. Interestingly, at least four of the upregulated genes were interferon-stimulated genes. Promoter analysis of the 2',5'-oligoadenylate synthetase gene, which was activated in both Tax-transformed Rat-1 cells and primary adult T-cell leukemia cells, demonstrated that Tax indirectly activates its interferon-responsive enhancer element in a nuclear factor-kappaB pathway-dependent manner, indicating a close association of interferon signaling with the transformation by Tax. PMID- 12650930 TI - Interaction between HERC1 and M2-type pyruvate kinase. AB - HERC proteins are characterized by having one or more RCC1-like domains as well as a C-terminal HECT domain in their amino acid sequences. This has led researchers to suggest that they may act as both guanine nucleotide exchange factors and E3 ubiquitin ligases. Here we describe a physical interaction between the HECT domain of HERC1, a giant protein involved in intracellular membrane traffic, and the M2 isoform of glycolytic enzyme pyruvate kinase (M2-PK). Partial colocalization of endogenous proteins was observed by immunofluorescence studies. This interaction neither induced M2-PK ubiquitination nor affected its enzymatic activity. The putative significance of the association is discussed. PMID- 12650931 TI - A new ligand for an old lipocalin: induced circular dichroism spectra reveal binding of bilirubin to bovine beta-lactoglobulin. AB - This study reports that bilirubin-bovine beta-lactoglobulin (BLG) complexes exhibit very characteristic induced circular dichroism (CD) spectra in the visible absorption region. Due to intramolecular chiral exciton coupling between the dipyrrinone chromophores, the long-wavelength negative and short-wavelength positive CD bands clearly prove that a single bilirubin molecule binds to BLG in a left-handed helical conformation (in pH 7.4 phosphate buffer Deltaepsilon(min) is -54 M(-1) cm(-1) at 467 nm and Deltaepsilon(max) is +48.5 M(-1) cm(-1) at 412 nm). The very low aqueous solubility and strong tendency of bilirubin molecules to aggregate around pH 7.4 meant that much more intense CD bands were measured at alkaline pH values owing to the increasing solubility of the ligand. Vanishing CD activity obtained upon titration of the complex with palmitic acid known to bind in the hydrophobic cavity of BLG indicates bilirubin to be bound at the open end mouth of the beta-barrel. Reversible changes of the induced CD spectrum due to acidic pH shift of the sample solution lead to the same conclusion. PMID- 12650932 TI - Voltammetry of a "protein on a rope". AB - A periplasmic electron-transfer protein, cytochrome c(555)(m) from Aquifex aeolicus contains a 62-residue N-terminal extension by which it is anchored to the membrane--most probably via a thioester bond to its N-terminal cysteine. This linker can act as a "rope" to tether the protein close to its reaction partners. Mimicking this principle, a recombinant cytochrome c(555)(m), expressed in Escherichia coli, has been attached covalently to a gold electrode modified with 6-mercaptohexan-1-ol. The "tethered" cytochrome c(555)(m) displays remarkably fast electron-transfer kinetics, with an electrochemical exchange rate constant k(0) of 1.4 x 10(4) s(-1). The results show that fast electron transfer is associated with weak interactions: importantly, the tethered cytochrome can explore many different orientations without escaping into solution. PMID- 12650933 TI - Thiamine biosynthesis in Escherichia coli: isolation and initial characterisation of the ThiGH complex. AB - In Escherichia coli, two of the proteins required for the biosynthesis of the thiazole moiety of thiamine (vitamin B(1)) are ThiG and ThiH, encoded as part of the thiCEFSGH operon. In this study, a C-terminally hexahistidine-tagged ThiH (ThiH-His) was expressed in E. coli as a soluble protein from thiGH-His-tag and thiFSGH-His-tag-bearing plasmids. When isolated under anaerobic conditions, ThiG and ThiH-His co-purify as a large multimeric non-covalent complex. Electron paramagnetic resonance and UV-visible spectroscopy together with iron and sulfide analyses revealed the presence of an iron-sulfur cluster within this complex. PMID- 12650934 TI - Detection of a concerted conformational change in the ATPase domain of DnaK triggered by peptide binding. AB - The molecular chaperone DnaK is composed of two functional domains, the ATPase domain and the substrate-binding domain. In this report, we show that peptide binding to DnaK can be sensed in real time through a labeled nucleotide bound in the ATPase domain. Specifically, when N8-(4-N'-methylanthraniloylaminobutyl)-8 aminoadenosine 5'-triphosphate (MABA)-ATP.DnaK complexes are rapidly mixed with excess peptide, MABA fluorescence rapidly increases and the rate of increase is proportional to peptide concentration. Analysis of the formation traces yield on and off rate constants that are exactly equal to the rate constants obtained from experiments that directly probe peptide binding to DnaK. These results are the first to show that peptide binding to ATP.DnaK triggers a concerted conformational change in the ATPase domain. PMID- 12650935 TI - The trichothecene biosynthesis gene cluster of Fusarium graminearum F15 contains a limited number of essential pathway genes and expressed non-essential genes. AB - We report for the first time the complete structure and sequence of the trichothecene biosynthesis gene cluster (i.e. Tri5-cluster) from Fusarium graminearum F15, a strain that produces 3-acetyldeoxynivalenol (3-ADON). A putative tyrosinase and polysaccharide deacetylase gene flank the Tri5-cluster: the number of pathway genes between them is less than half the total number of steps necessary for 3-ADON biosynthesis. In comparison with partial Tri5-cluster sequences of strains with 15-acetyldeoxynivalenol and 4-acetylnivalenol chemotypes, the Tri5-cluster from strain F15 contains three genes that are apparently unnecessary for the biosynthesis of 3-ADON (i.e. Tri8 and Tri3, which are expressed, and pseudo-Tri13, which is not expressed). In addition, the Tri7 gene was missing from the cluster. Recombinant TRI3 protein showed limited trichothecene C-15 acetylase activity. In contrast, recombinant TRI8 protein displayed no C-3 deacetylase activity, suggesting that the loss or alteration of function contribute directly to the chemotype difference. PMID- 12650936 TI - Gene silencing by adenovirus-delivered siRNA. AB - RNA interference is the process that double-stranded RNA induces the homology dependent degradation of cognate mRNA mediated by 21-23 nucleotide short interfering RNA (siRNA). Here, we describe a simple virus vector for efficient delivery of siRNA into mammalian cells utilizing the well-defined H1-RNA promoter and conventional adenovirus. In this pilot study, p53 was targeted by this vector. Our results demonstrate efficient and specific knock-down of p53 in breast cancer MCF-7 and lung carcinoma A549 cells and indicate a prospective application of this siRNA expressing recombinant adenovirus system in functional genomics, cancer gene therapy and virus inhibition. PMID- 12650937 TI - A cell shrinkage-induced non-selective cation conductance with a novel pharmacology in Ehrlich-Lettre-ascites tumour cells. AB - In whole-cell recordings on Ehrlich-Lettre-ascites tumour (ELA) cells, the shrinkage-induced activation of a cation conductance with a selectivity ratio P(Na):P(Li):P(K):P(choline):P(NMDG) of 1.00:0.97:0.88:0.03:0.01 was observed. In order of potency, this conductance was blocked by Gd(3+)=benzamil>amiloride>ethyl isopropyl-amiloride (EIPA). In patch-clamp studies using the cell-attached configuration, a 14 pS channel became detectable that was reversibly activated upon hypertonic cell shrinkage. It is concluded that ELA cells express a shrinkage-induced cation channel that may reflect a molecular link between amiloride-sensitive and -insensitive channels. In addition, because of its pharmacological profile, it may possibly be related to epithelial Na+ channels (ENaCs). PMID- 12650938 TI - Structural characterization of thyroglobulin type-1 domains of equistatin. AB - Equistatin is a protein composed of three thyroglobulin type-1 domains. It inhibits papain-like cysteine proteinases and the aspartic proteinase, cathepsin D. To determine the structural basis for this inhibition we cloned and expressed the separated domains (eq d-1, eq d-2, eq d-3) in Pichia pastoris. Kinetic constants for the interaction of eq d-1 with papain and that of eq d-2 with cathepsin D are of similar order (subnanomolar) and are comparable to the constants obtained for full-length equistatin. The target proteinase for the third domain remains unknown. Thus, we demonstrate here that thyroglobulin type-1 motifs per se are able to support specific structural features that enable them to inhibit proteases from different classes. The overall conformation of three domains in equistatin is such that the interaction of domains 1 or 2 with their respective target enzymes is not hindered sterically by either domain. In addition, we show that the interaction of eq d-2 with cathepsin D results in conformational changes, which is not the case for the eq d-1/papain interaction. PMID- 12650939 TI - Structural cooperativity in the SH3 domain studied by site-directed mutagenesis and amide hydrogen exchange. AB - We have studied the effects produced by site-directed mutagenesis upon energetic and structural cooperativity in the Src homology region 3 domain of alpha spectrin. The mutation of Asn47 to Gly or Ala in the distal loop brings about significant changes to the global stability of the domain in spite of not affecting its structure to any great extent. The binding affinity for a proline rich peptide is also largely diminished in both mutant domains. We have compared the apparent Gibbs energies of the amide hydrogen-deuterium exchange (HX) between the wild-type and the Gly47 mutant. The observed changes in the Gibbs energy of HX indicate a remarkable energetic cooperativity in this small domain. Regions of the domain's core have a high cooperativity with the position of the mutation, indicating that their HX occurs mainly in states in which the distal loop is unstructured. More flexible regions, which undergo HX mainly by local motions, show a lower but still considerable cooperativity with the distal loop. We conclude that there is an important correlation between regional stability and cooperativity in this small domain. PMID- 12650940 TI - ArhGAP15, a novel human RacGAP protein with GTPase binding property. AB - We have previously described a partial cDNA sequence encoding a RhoGAP protein, GAP25 that is homologous to the recently reported ArhGAP9 and ArhGAP12. We now describe a related new member ArhGAP15 that shares a number of domain similarities, including a pleckstrin homology (PH) domain, a RhoGAP domain and a novel motif N-terminal to the GAP domain. This novel motif was found to be responsible for nucleotide-independent Rac1 binding. Using swop mutants of Rac/Cdc42, we have established that the binding is through the C-terminal half of Rac1. The GAP domain of ArhGAP15 showed specificity towards Rac1 in vitro. The PH domain is required for ArhGAP15 to localize to cell periphery and over-expression of the full-length ArhGAP15, but not the mutant with a partial deletion of the PH domain, resulted in an increase in actin stress fibers and cell contraction. These morphological effects can be attenuated by the co-expression of dominant negative Rac1(N17). HeLa cells expressing ArhGAP15 were also resistant to phorbol myristatate acetate treatment, suggesting that ArhGAP15 is a potential regulator of Rac1. PMID- 12650941 TI - Solution structure of CnErg1 (Ergtoxin), a HERG specific scorpion toxin. AB - The three-dimensional structure of chemically synthesized CnErg1 (Ergtoxin), which specifically blocks HERG (human ether-a-go-go-related gene) K+ channels, was determined by nuclear magnetic resonance spectroscopy. CnErg1 consists of a triple-stranded beta-sheet and an alpha-helix, as is typical of K+ channel scorpion toxins. The peptide structure differs from the canonical structures in that the first beta-strand is shorter and is nearer to the second beta-strand rather than to the third beta-strand on the C-terminus. There is also a large hydrophobic patch on the surface of the toxin, surrounding a central lysine residue, Lys13. We postulate that this hydrophobic patch is likely to form part of the binding surface of the toxin. PMID- 12650942 TI - Truncated thioredoxin (Trx80) exerts unique mitogenic cytokine effects via a mechanism independent of thiol oxido-reductase activity. AB - Recently we discovered that a naturally occurring C-terminally truncated thioredoxin (Trx80) is a potent mitogenic cytokine stimulating IL-12 production from CD40(+) monocytes. To further characterise Trx80 we have engineered cysteine to serine mutants of Trx80 corresponding to the active site cysteines of Trx (Trx80SGPS) and to the structural cysteine at position 72 (Trx80C72S). Trx80SGPS and Trx80C72S retained the cell stimulatory activity of Trx80 and increased peripheral blood mononuclear cell (PBMC) proliferation three- to five-fold in vitro (P<0.01, n=18). Both Trx80SGPS and Trx80C72S significantly stimulated IL-12 and IFN-gamma secretion from PBMCs in the same manner as Trx80 (P<0.01, n=9 and 10). The previously described Trx80 dimer is caused by non-covalent interactions, and not by any intermolecular disulphide bonds. PMID- 12650943 TI - Recombinant endostatin forms amyloid fibrils that bind and are cytotoxic to murine neuroblastoma cells in vitro. AB - Endostatin is a fragment of collagen XVIII that acts as an endogenous inhibitor of tumor angiogenesis and tumor growth. Anti-tumor effects have been described using both soluble and insoluble recombinant endostatin. However, differences in endostatin structure are likely to cause differences in bioactivity. In the present study we have investigated the structure and cellular effects of insoluble endostatin. We found that insoluble endostatin shows all the hallmarks of amyloid aggregates. Firstly, it binds Congo red and shows the characteristic apple-green birefringe when examined under polarized light. Secondly, electron microscopy shows that endostatin forms short unbranched fibrils. Thirdly, X-ray analysis shows the abundant presence of cross-beta sheets, the tertiary structure that underlies fibrillogenesis. None of these properties was observed when examining soluble endostatin. Soluble endostatin can be triggered to form cross beta sheets following denaturation, indicating that endostatin is a protein fragment with an inherent propensity to form amyloid deposits. Like beta-amyloid, found in the brains of patients with Alzheimer's disease, amyloid endostatin binds to and is toxic to neuronal cells, whereas soluble endostatin has no effect on cell viability. Our results demonstrate a previously unrecognized functional difference between soluble and insoluble endostatin, only the latter acting as a cytotoxic amyloid substance. PMID- 12650944 TI - A new human gene KCNRG encoding potassium channel regulating protein is a cancer suppressor gene candidate located in 13q14.3. AB - We report the primary characterization of a new gene KCNRG mapped at chromosome band 13q14.3. This gene includes three exons and has two alternatively spliced isoforms that are expressed in normal tissues and in some tumor cell lines. Protein KCNRG has high homology to tetramerization domain of voltage-gated K+ channels. Using the patch-clamp technique we determined that KCNRG suppresses K+ channel activity in human prostate cell line LNCaP. It is known that selective blockers of K+ channels suppress lymphocyte and LNCaP cell line proliferation. We suggest that KCNRG is a candidate for a B-cell chronic lymphocytic leukemia and prostate cancer tumor suppressor gene. PMID- 12650945 TI - Modeling and mutational analysis of the GAF domain of the cGMP-binding, cGMP specific phosphodiesterase, PDE5. AB - The GAFa domain of the cGMP-binding, cGMP-specific phosphodiesterase (PDE5A) was modeled on the crystal structure of PDE2A GAF domain and residues involved in cGMP binding identified. Tandem GAFa and GAFb domains of PDE5A, expressed in Escherichia coli, bound cGMP (K(d) 27 nM). Mutation of aspartate-299 in GAFa, suggested earlier to be critical for cGMP binding, did not abrogate cGMP binding, but mutation of F205, which formed a stacking interaction with the guanine ring of cGMP, led to complete loss of cGMP binding. Therefore, the GAFa domain of PDE5A adopts a structure similar to the GAFb domain of PDE2A, and provides the sole site for cGMP binding in PDE5A. PMID- 12650946 TI - Identification of three novel Smad binding proteins involved in cell polarity. AB - A yeast two-hybrid screen was utilized to identify novel Smad 3 binding proteins expressed in developing mouse orofacial tissue. Three proteins (Erbin, Par-3, and Dishevelled) were identified that share several similar structural and functional characteristics. Each contains at least one PDZ domain and all have been demonstrated to play a role in the establishment and maintenance of cell polarity. In GST (glutathione S-transferase) pull-down assays, Erbin, Par-3, and Dishevelled bound strongly to the isolated MH2 domain of Smad 3, with weaker binding to a full-length Smad 3 protein. Failure of Erbin, Par-3, and Dishevelled to bind to a Smad 3 mutant protein that was missing the MH2 domain confirms that the binding site resides within the MH2 domain. Erbin, Par-3, and Dishevelled also interacted with the MH2 domains of other Smads, suggesting broad Smad binding specificity. Dishevelled and Erbin mutant proteins, in which the PDZ domain was removed, still retained their ability to bind Smad 3, albeit with lower affinity. While transforming growth factor beta (TGFbeta) has been suggested to alter cell polarity through a Smad-independent mechanism involving activation of members of the RhoA family of GTP binding proteins, the observation that Smads can directly interact with proteins involved in cell polarity, as shown in the present report, suggests an additional means by which TGFbeta could alter cell polarity via a Smad-dependent signaling mechanism. PMID- 12650947 TI - The pharmacology of putative early-onset antidepressant strategies. AB - Depression is a serious and burdensome illness. Although selective serotonin reuptake inhibitors (SSRIs) have improved safety and tolerability of antidepressant treatment efficacy, the delay in the onset of action have not been improved. There is evidence to suggest that the delay in onset of therapeutic activity is a function of the drugs, rather than the disease. This suggests that research into the biological characteristics of depression and its treatments may yield faster-acting antidepressants. Emerging evidence from clinical studies with mirtazapine, venlafaxine and SSRI augmentation with pindolol suggests that these treatments may relieve antidepressant symptoms more rapidly than SSRIs. The putative mechanism of action of faster-acting antidepressant strategies presented here purports that conventional antidepressants acutely increase the availability of serotonin (5-hydroxytryptamine, 5-HT) or noradrenaline (NA), preferentially at their cell body level, which triggers negative feedback mechanisms. After continued stimulation, these feedback mechanisms become desensitised and the enhanced 5-HT availability is able to enhance 5-HT and/or NA neurotransmission. Putative fast-onset antidepressants, on the other hand, may uncouple such feedback control mechanisms and enhance 5-HT and/or NA neurotransmission more rapidly. Further studies are required to characterise in detail the interactions between NA and 5-HT systems and to definitively establish the early onset of candidate antidepressants such as mirtazapine, venlafaxine and pindolol augmentation. PMID- 12650948 TI - Serum iron levels in schizophrenic patients with or without akathisia. AB - The pathophysiology of akathisia still remains controversial. Iron deficiency was proposed to be an important factor in the development of akathisia. In the present study, it was aimed to compare levels of serum iron and linked variables in chronic akathisic (n=30), and non-akathisic patients (n=30) with schizophrenia and healthy controls (n=30) because of the controversy in the association of iron and akathisia. The Barnes Akathisia Scale for akathisia and Simpson-Angus Rating Scale for extrapyramidal side effects were used. Serum iron and linked variables and hematological profile of the patients and control subjects were determined. Serum iron levels were significantly lower both in akathisic and non-akathisic groups compared to the control group (P<0.001). Moreover, akathisic patients had significantly lower iron levels than non-akathisic patients (P<0.05). Total iron binding capacity was significantly higher in patients with akathisia compared to the control group (P<0.01). Although non-akathisic patients had a mild increase in total iron binding capacity, it was not statistically significant compared to the control group (P>0.05). Ferritin levels were determined to be significantly lower in both groups compared to the control group (P<0.01). In addition, there was a significant difference in ferritin levels between the patients with and without akathisia (P<0.05). In conclusion, our results support the hypothesis that an association between akathisia and iron metabolism exists. PMID- 12650949 TI - Milnacipran in the treatment of bulimia nervosa: a report of 16 cases. AB - Controlled trials in patients with bulimia nervosa have demonstrated efficacy of antidepressant medications with serotonergic function (e.g. fluoxetine) as well as noradrenergic function (e.g. desipramine). Sixteen out-patients with bulimia nervosa according to DSM-IV criteria were treated in a drug surveillance with 100 mg of milnacipran, a specific serotonin and noradrenaline reuptake inhibitor (SNRI). Ten patients completed the 8-week observation period. The reasons for premature attrition were improvement in one patient (no. 12), a generalized exanthema in one patient (no. 7), severe nausea in one patient (no. 8) and non compliance due to non-drug-related reasons in three patients (no. 1, 2, and 16). An intent-to-treat analysis exhibited a significant reduction in weekly binge eating and vomiting frequency from baseline to the end of treatment. Three patients stopped binge eating and purging completely during the last week of treatment. Furthermore, there was a concomitant decrease of depression ratings (HAMD, BDI). Our preliminary data give rise to the notion that milnacipran may be promising in the treatment of bulimia nervosa. PMID- 12650951 TI - Alternation learning in OCD/schizophrenia patients. AB - In continuation of our previous studies on orbitofrontal cortex function in obsessive compulsive disorder (OCD) we studied a group of OCD patients with schizophrenia in comparison with a group of schizophrenia patients. In order to test orbitofrontal cortex function we again used an alternation learning task. We found no difference between the two groups in the performance of this task. The relationship between severity of OC symptoms and alternation learning was curvilinear. Thus, in the low range of symptom severity the correlation between alternation learning and symptom severity was negative, while in the high range it was positive. The positive correlation in the severely affected patients is essentially the same as that found in severe "pure" OCD patients, which we have previously reported. We conclude that the data indicate independent orbitofrontal cortex function in OCD, irrespective of comorbidity. PMID- 12650950 TI - Nizatidine for prevention of weight gain with olanzapine: a double-blind placebo controlled trial. AB - Weight gain is associated with treatment with olanzapine and other psychotropic agents. Nizatidine, a histamine H-2 receptor antagonist, has been proposed to have weight-reducing effects. This double-blind trial evaluated the efficacy of nizatidine in limiting weight gain in patients with schizophrenia and related disorders who were treated with olanzapine for up to 16 weeks. After an initial screening period, 175 patients were randomized to receive olanzapine (5-20 mg) with either placebo or nizatidine (150 mg b.i.d. or 300 mg b.i.d.). Significantly less weight gain was observed on average at weeks 3 and 4 with olanzapine+nizatidine 300 mg b.i.d. (P<0.05) compared to olanzapine+placebo, but the difference was not statistically significant at 16 weeks. Nizatidine was well tolerated and did not adversely affect clinical outcomes. Nizatidine 300 mg b.i.d. may have an early transient effect in limiting the weight gain, but this potential early effect appeared to be diminished or eliminated by 16 weeks. PMID- 12650952 TI - Investigating the role of dopaminergic and serotonergic candidate genes in obsessive-compulsive disorder. AB - There is increasing evidence that the aetiology of obsessive-compulsive disorder (OCD) has a marked genetic component, although the precise mechanism of inheritance is unclear. Clinical and pharmacological studies have implicated the serotonergic and dopaminergic systems in disease pathogenesis. This study investigated the role of attractive candidate genes in the serotonergic and dopaminergic pathways in the development of OCD. The distribution of selected polymorphic variants in the serotonin receptor type 2A and 1Dbeta (5-HT(2A), 5 HT(1Dbeta)), dopamine transporter (DAT), dopamine receptor type 4 (DRD4) and monoamine-oxidase A (MAO-A) genes were analysed in 71 OCD cases and 129 control individuals in the genetically homogeneous Afrikaner population, by means of case control association studies. Although no statistically significant genotypic or allelic associations were detected, the data yielded interesting preliminary results that warrant further discussion and investigation. PMID- 12650953 TI - Polyunsaturated fatty acid deficit in patients with bipolar mania. AB - The aim of this study is to test the hypothesis that there is a depletion of polyunsaturated fatty acids of erythrocyte membranes in patients with bipolar disorder and to connect the previous therapeutic and psychoimmunological findings. Fatty acid compositions of erythrocyte membranes in 20 bipolar manic patients and 20 healthy controls were analyzed by thin-layer chromatography and gas chromatography. The major finding was significantly reduced arachidonic acid (20:4n-6) and docosahexaenoic acid (22:6n-3) compositions in bipolar patients as compared to normal controls with P values of 0.000 and 0.002, respectively. There were no differences in total omega-3 and omega-6 polyunsaturated fatty acids. This abnormality may be related to the mechanisms of action of mood stabilizers and the previous findings on the abnormal psychoimmunology of patients with bipolar disorder. Larger sample sizes of medicated patients or drug-free manic, well-controlled designs on the diet and smoking, and fatty acid composition measurements during full remission after the index episode are warranted in future studies. PMID- 12650954 TI - CSF testosterone in 43 male suicide attempters. AB - Several studies have shown a relationship between high testosterone and violent aggressive behaviour. The general aim of this study was to gain knowledge of the importance of testosterone in suicide attempters. Testosterone in cerebrospinal fluid (CSF) was analysed in men with a recent suicide attempt, diagnostically subdivided into groups according to DSM-III-R axis I and II diagnosis and mode of suicidal behaviour. In general, our patients had lower CSF testosterone levels than aggressive violent patients in other studies. Patients with depression NOS or dysthymia showed higher CSF testosterone levels than the rest. Significant positive correlation between testosterone and irritability or a negative correlation with social desirability was found in diagnostic subgroup of patients, specifically axis II, cluster B personality disorders. The results suggest that suicide attempts may be mediated by different biological variables than aggression. PMID- 12650955 TI - Effects of zolpidem 10 mg, zopiclone 7.5 mg and flunitrazepam 1 mg on night-time motor activity. AB - The effects of a single dose of zolpidem (10 mg), zopiclone (7.5 mg) and flunitrazepam (1 mg) on motor activity the following 3 nights were compared to those of a placebo in a double-blind, crossover study. Thirty-three healthy subjects received medication between 10.30 and 11.30 p.m. and were asked to rise between 7.30 and 8.30 a.m. During the night under treatment, flunitrazepam, zopiclone and zolpidem significantly reduced motor activity. Changes in motor activity are quantitatively compatible with the hypothesis of reduced light sleep and wakefulness after sleep onset. During the first or second post-drug night, for zolpidem and zopiclone the opposite effect was observed, i.e. increased activity compared with placebo. These modifications cannot be explained by modified sleep structure. This last result underlines our inadequate understanding of the underlying mechanisms of motor activity during sleep. However, being sensitive and easy to use, actigraphy is an ideal technique to assess the effect of hypnotics on large populations and for long duration studies. PMID- 12650956 TI - SSRIs antidepressant activity is influenced by G beta 3 variants. AB - The aim of the present study was to test a possible effect of the G-protein beta3 subunit (Gbeta3) C825T gene variant on the antidepressant activity of selective serotonin reuptake inhibitors (SSRIs) in a sample of major and bipolar depressives, with or without psychotic features. Four hundred and ninety inpatients were treated with fluvoxamine 300 mg/day (n=362) or paroxetine 40 mg/day (n=128) and either placebo or pindolol in a double-blind design for 6 weeks. The severity of depressive symptoms was weekly assessed with the Hamilton Rating Scale for Depression. Gbeta3 allelic variants were determined in each subject using a PCR-based technique. Subjects with Gbeta3 T/T variants showed better response to treatment (P=0.009) and this effect was independent from analyzed demographic and clinical variables. These results confirm preliminary reports and shed further light on the genetics of the response to antidepressant treatments. PMID- 12650957 TI - Effect of the 5-HT2 antagonist mianserin on cognitive dysfunction in chronic schizophrenia patients: an add-on, double-blind placebo-controlled study. AB - Preponderance of serotonin 5-HT2A antagonism over dopamine D2 blockade exerted by atypical antipsychotics may contribute to their cognitive-enhancing effect. In a double-blind placebo-controlled study we examined the effect of add-on mianserin (15 mg/day), an agent with marked 5-HT2A antagonism, on cognitive functioning in 30 chronic hospitalized DSM-IV schizophrenia patients stabilized on typical antipsychotics. The Automated Neuropsychological Assessment Metrics (ANAM) battery was used to assess learning, memory and sustained attention; Wisconsin Card Sorting Test (WCST) to assess executive function at baseline and endpoint (4 weeks). Clinical assessment included appropriate rating scales. The mianserin group overperformed the placebo group on selective ANAM memory/learning tests, reflected in moderate-to-high effect size values. No between-group differences were revealed in WCST and clinical ratings. CONCLUSIONS: Improved performance on selective neurocognitive tests with addition of the 5-HT2A antagonist mianserin to typical antipsychotics indicates a possible role of the 5-HT system in cognitive-enhancing effects. The effect of flexible doses of mianserin on cognitive deficits in a broader schizophrenia population merits further investigation. PMID- 12650958 TI - Clonazepam in the long-term treatment of patients with unipolar depression, bipolar and schizoaffective disorder. AB - The value of a long-term treatment with clonazepam in the prophylaxis of affective disorder is discussed controversially in the scientific literature. Altogether there are only a few reports on the use of this compound as a mood stabilizer, most of them describing patients suffering from bipolar affective disorder. The aim of this investigation was to evaluate clonazepam as a phase prophylactic medication in affective disorder. We conducted a retrospective chart review in 34 out-patients of our lithium clinic (15 suffering from unipolar depression, 15 from bipolar disorder, four from schizoaffective disorder), who had been treated with clonazepam as a long-term medication. Clonazepam was either given as monotherapy, or as in the case of lithium non-responders, as adjunctive therapy. Patients with unipolar depression had significantly (P=0.026) less depressive episodes after initiation of treatment with clonazepam. Patients with bipolar disorder did not benefit from this therapy. Neither manic/hypomanic phases nor depressive episodes were reduced in this group of patients. Interestingly, clonazepam also reduced affective phases in our four schizoaffective patients on a trend level. Our results indicate that patients with unipolar depression may benefit from a maintenance treatment with clonazepam. Due to methodological limitations our results need to be replicated in controlled double-blind randomized clinical trials. PMID- 12650959 TI - Manic symptoms associated with quetiapine treatment. AB - This case illustrates the induction of manic-like symptoms in a 26-year-old male patient with DSM-IV paranoid schizophrenia following treatment with quetiapine. The only drug he had received prior to quetiapine was risperidone which was occasionally taken in the previous 3 years. The manic symptoms remitted after quetiapine withdrawal. PMID- 12650960 TI - Upregulated iNOS and oxidative damage to the cochlear stria vascularis due to noise stress. AB - Our previous work has revealed increased nitric oxide (NO) production in the cochlear perilymph following noise stress. However, it is not clear if the increase of NO is related to iNOS and whether NO-related oxidative stress can cause vascular tissue damage. In this study, iNOS immunoreactivity, NO production, and reactive oxygen species (ROS) in the lateral wall were examined in normal mice and compared with similar animals exposed to 120 dBA broadband noise, 3 h/day, for 2 consecutive days. In the normal animals, iNOS expression was not observed in the vascular endothelium of the stria vascularis and only weak iNOS immunoactivity was detected in the marginal cells. However, expression of iNOS in the wall of the blood vessels of stria vascularis and marginal cells was observed after loud sound stress (LSS). Relatively low levels of NO production and low ROS activity were detected in the stria vascularis in the unstimulated condition. In contrast, NO production was increased and ROS activity was elevated in the stria vascularis after LSS. These changes were attenuated by the iNOS inhibitor, GW 274150. To explore whether noise induces apoptotic processes in the stria vascularis, we examined morphological changes in endothelial- and marginal-cells. In vitro, annexin-V phosphatidylserine (PS) (to label and detect early evidence of apoptosis) was combined with propidium iodide (PI) (to probe plasma membrane integrity). PI alone was used in fixed tissues to detect later stage apoptotic cells by morphology of the nuclei. Following LSS, PS was expressed on cell surfaces of endothelial cells of blood vessels and marginal cells of the stria vascularis. Later stage apoptosis, characterized by irregular nuclei and condensation of nuclei, was also observed in these cells. The data indicate that increased iNOS expression and production of both NO and ROS following noise stress may lead to marginal cell pathology, and the dysfunction of cochlear microcirculation by inducing blood vessel wall damage. PMID- 12650961 TI - Identification of a key protein associated with cerebral ischemia. AB - The biochemical effects of permanent focal ischemia following unilateral occlusion of the middle cerebral artery in rats were studied by determining the content of specific proteins of the affected areas in the cerebral hemisphere. Brain proteins were prepared 72 h after the occlusion and analyzed by sodium dodecylsulfate-polyacrylamide gel electrophoresis. A significant increase in 66 and 80 kDa components and a paradoxical decrease in 260 kDa protein occurred in the ischemic brain tissues. The 66 and 80 kDa protein bands were identified as albumin and transferrin, respectively. The 260 kDa protein was analyzed by peptide mass fingerprinting (PMF) and matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The isoelectric point of the 260 kDa protein was 4.65 determined by isoelectric focusing. The data obtained from PMF were used in searching the protein database for homologous components. Three proteins with partial homology were identified. They were the microtubule associated protein 1A, protein-tyrosine phosphatase zeta precursor (phosphacan), and protein kinase A anchoring protein 6. Polyclonal antibodies against the 260 kDa protein were raised and used to immunolocalize the antigen in various tissues. Positive staining occurred with brain neurons and pyramidal cells, islet cells, podocytes of kidney glomeruli, and endothelial cells of the venous sinuses of the spleen. The localization of 260 kDa protein strongly implies its function in these tissues. Its physiological and pathophysiological significances need to be clarified in future. PMID- 12650962 TI - Light-induced photoreceptor degeneration may involve the NF kappa B/caspase-1 pathway in vivo. AB - PURPOSE: To determine the role of nuclear factor-kappaB (NFkappaB) in light induced photoreceptor degeneration. METHODS: Dark-adapted BALB/cJ mice, 4-8 weeks, were exposed to an intense green light (3.1-3.5 klux) for 1, 3, 6, 9, 12, or 24 h and killed immediately after exposure. The photoreceptor apoptosis was detected by TUNEL. Co-localization of NFkappaB p65 immunoreactivity and TUNEL in photoreceptor cells was detected by double immunolabeling. The protein levels of X-linked inhibitor of apoptosis protein (XIAP), Bcl-xL, caspase-1, and opsin after light exposure were analyzed by Western blot analysis. In addition, the initiation of NFkappaB activation was assessed by measuring the increase in phosphorylated IkappaBalpha (pIkappaBalpha). Immunohistochemical localization of caspase-1 was also performed on the mouse retinas. RESULTS: Co-localization of NFkappaB p65 immunoreactivity with TUNEL was observed in scattered photoreceptor cells after 24 h of light exposure. The amount of pIkappaBalpha was increased after 1 h of light exposure, and in parallel, the amounts of XIAP and Bcl-xL were increased at 1 h. In contrast, caspase-1 did not increase until after 6 h of light exposure. Caspase-1-immunolabeling was observed in scattered photoreceptor cells after 3 h of light exposure but was markedly increased in many more cells at 6 h. CONCLUSIONS: These findings suggest that NFkappaB may play an anti apoptotic role in the early response to light stress and that photoreceptor apoptosis induced by light stress may be mediated through an NFkappaB/caspase-1 pathway. PMID- 12650963 TI - Inhibition of L-type voltage dependent calcium channels causes impairment of long term potentiation in the hippocampal CA1 region in vivo. AB - Long-term potentiation (LTP), in the hippocampal CA1 region is dependent on postsynaptic calcium influx. It is generally accepted that calcium influx occurs via activation of the NMDA receptor channel complex. However, studies in vitro using a high-frequency stimulus protocol (> or =200 Hz) demonstrated previously an NMDA receptor-independent form of LTP that is dependent upon activation of L type voltage-dependent calcium channels (VDCCs). Here we have investigated a role for L-type VDCCs in LTP in vivo. Two structurally different, L-type VDCC blockers, verapamil (1, 3 and 10 mg/kg) and diltiazem (1, 10 and 20 mg/kg), depressed the induction of LTP in a dose-dependent manner. Increased activation of L-type VDCCs by Bay K 8644, an L-type agonist, however, did not enhance LTP. The NMDA receptor antagonist D-AP5 (5 and 20 mM injected i.c.v) impaired, but failed to block fully LTP in vivo. A reduced level of LTP could still be recorded following co-administration of verapamil and D-AP5. The level of LTP recorded was similar to that observed in the presence of either verapamil (10 mg/kg) or D-AP5 alone. These results suggest that activation of the NMDA receptor/channel and L type VDCCs are involved in the induction of LTP in area CA1 in vivo. However, it appears that activation of other receptor/channels may also play a role in this form of LTP. PMID- 12650964 TI - Immunoelectron microscopic localization of the opioid growth factor receptor (OGFr) and OGF in the cornea. AB - This study was conducted to determine the cellular and subcellular location(s) of the opioid growth factor receptor (OGFr), and the opioid growth factor (OGF), [Met(5)]-enkephalin, in the corneal epithelium. Laser scanning confocal microscopy analysis revealed that both OGFr and OGF were colocalized in the paranuclear cytoplasm and cell nuclei in basal, as well as suprabasal, cells of adult rat corneal epithelium. Using a postembedding immunogold procedure for immunoelectron microscopy that included embedding in Unicryl, both single- and double-face labeling studies were performed. Immunogold labeling of OGFr was detected on the outer nuclear envelope, in the paranuclear cytoplasm proximal to the nuclear envelope, perpendicular to the nuclear envelope in a putative nuclear pore complex, and within the nucleus adjacent to heterochromatin. Immunoreactivity for OGF was noted in locations similar to that for OGFr. In addition, aggregates of staining for OGF were found throughout the cytoplasm, including subjacent to the plasma membrane. Double labeling experiments revealed that complexes of OGF-OGFr were colocalized on the outer nuclear envelope, in the paranuclear cytoplasm, extending across the nuclear pore complex, and in the nucleus. Anti-OGFr IgG by itself, but not anti-OGF IgG alone, was associated with the outer nuclear envelope, and uncomplexed OGF immunoreactivity was detected in the cytoplasm in dual labeling experiments. These results based on complementary approaches of confocal microscopy and immunoelectron microscopy, suggest that: (i) OGFr resides on the outer nuclear envelope, (ii) OGF interacts with OGFr at the outer nuclear envelope, (iii) the colocalized receptor and peptide translocates between the cytoplasm and the nucleus at the nuclear pore, and (iv) signal transduction for modulation of cell proliferation necessitates a peptide receptor complex that interfaces with chromatin in the nucleus. PMID- 12650965 TI - Organization of the circadian system in the subterranean mole rat, Cryptomys hottentotus (Bathyergidae). AB - The mole rat, Cryptomys hottentotus (Bathyergidae) is a gregarious subterranean rodent, which shows no entrainment to ambient light-dark cycles. The locomotor activity of individuals or of a whole colony, which shows no circadian rhythmicity. Since the lack of both synchronization to light-dark cycle and an endogenous rhythm of locomotor activity could be related to the organization of the circadian system, we have investigated the neuropeptidergic features of the SCN and IGL, and have used pseudorabies viral tracing methods to identify the visual and circadian pathways in this species. The precise topographic distribution of certain neuropeptide populations in the SCN differs from typical rodent pattern of organization and may be correlated with the apparent absence of light entrainment of activity and lack of endogenous rhythmicity. The IGL is highly reduced in size. This structure can nevertheless be identified by the presence of NPY and CALB positive neurons, as well as by a dense network of SP fibers. Viral tracing using intraocular injection of the PRV-Becker and PRV Bartha strains, leads to differential infection of neurons in circadian and visual structures. With the Bartha strain, infected neurons are principally observed in the SCN, whereas the Becker strain leads primarily to infection of the dLGN and shows an anatomical regression of visual structures. Transsynaptic retrograde infection of the retina contralateral to the injected eye reveals a morphologically homogeneous population, which resemble to retinohypothalamic ganglion cells described in other mammals. PMID- 12650966 TI - Testosterone-dependent and -independent mechanisms involved in the photoperiodic control of neuropeptide levels in the brain of the jerboa (Jaculus orientalis). AB - Vasoactive intestinal peptide (VIP) and neuropeptide Y (NPY) content in the suprachiasmatic nucleus have been shown to exhibit seasonal changes with an increase in late summer, the period of sexual quiescence in the jerboa (Jaculus orientalis). In this study, VIP content in the SCN and NPY and enkephaline (ENK) content in the geniculo-suprachiasmatic system have been assayed in wild-caught male jerboas (Jaculus orientalis) in order to determine whether these neuropeptides are controlled directly by photoperiod changes or indirectly by short photoperiod induced changes in circulating sex hormones levels. In agreement with previous studies seasonal variations occur in the VIP and NPY content in the SCN. Variations also occur in NPY content in the IGL with an increase in the period of sexual quiescence. In contrast, no seasonal changes are observed in Enk content in the IGL or the SCN. In short photoperiod conditions increases are observed in both VIP and NPY content in the SCN as well as NPY content in the IGL. Castration during the period of sexual activity (spring) or under long photoperiod which drastically reduces testosterone, also induced an increase in the levels of these neuropeptides. Testosterone implants which reproduce the sex hormonal status of the sexual activity period failed to prevent the short photoperiod-induced increase of VIP and NPY in the SCN and of NPY in the IGL. These results clearly show that the photoperiod modulates VIP and NPY in the geniculo-suprachiasmatic system both by testosterone-linked and testosterone independent mechanisms. PMID- 12650967 TI - Nitrous oxide-induced c-Fos expression in the rat brain. AB - Induction of c-Fos has previously been used to map locations of cells in the central nervous system (CNS) that are activated by ethanol administration. Only a few studies examining a restricted range of CNS areas have identified brain areas activated by nitrous oxide (N(2)O). Because ethanol and N(2)O have overlapping physiological, psychological and behavioral effects, we hypothesized that these drugs act on similar sites in the CNS. To test this hypothesis, we assessed c-Fos like immunoreactivity in brain slices from male Long-Evans rats that received a 2 h exposure of 0, 20, 40 or 60% N(2)O (n=5 each) immediately prior to sacrifice. N(2)O administration produced significant (P<0.05) dose-related increases of c Fos expression in several forebrain regions, including the hypothalamic supraoptic and paraventricular nuclei, the thalamic paraventricular nucleus, the amygdala, and in retrosplenial cortex. In the midbrain, N(2)O caused significant dose-related c-Fos expression in the Edinger-Westphal nucleus. Finally, the pontine locus coeruleus, and two medullary regions, the nucleus of the solitary tract and ventrolateral medulla, also showed significant dose-related N(2)O induced c-Fos expression. Most of the brain areas identified as targets of N(2)O are also activated by ethanol administration. The overlapping pattern of c-Fos induced by ethanol and N(2)O suggests that these drugs may cause comparable central activity by acting on similar neuronal pathways. PMID- 12650968 TI - Chemical bladder irritation provokes c-fos expression in the midbrain periaqueductal gray matter of the rat. AB - We investigated the topographical localization of c-fos expression in the midbrain periaqueductal gray matter (PAG) to detect nociception-induced neural activity in the PAG. In conscious female Wistar rats, c-fos expression was induced by continuous intravesical infusion of saline or 0.1% acetic acid. Number of c-fos protein (Fos)-positive cells was counted at each coronal section of the PAG as well as Barrington's nucleus. Fos-positive cells were also counted at L1 and L6 of the spinal cord, where most of the hypogastric and pelvic nerve afferent terminals project, respectively. Compared with saline infusion, acetic acid infusion provoked irritative bladder responses characterized by a marked increase in the frequency of bladder contractions, and induced a significant increase in the number of Fos-positive cells in both L1 and L6 of the spinal cord. Following acetic acid infusion, there was a significant increase in the number of Fos-positive cells in all coronal sections of the PAG compared with saline infusion, especially in the caudal part of the PAG. The increase in the number of Fos-positive cells was mainly observed in the ventrolateral and lateral parts of the caudal PAG, and in the dorsal part of the rostral PAG. However, there was no difference in the number of Fos-positive cells in Barrington's nucleus between saline and acetic acid infusion. In conclusion, nociception induced by chemical bladder irritation influences neural activity in the PAG. Implication of topographical difference in Fos expression in the PAG and its relevance to changes in bladder function remain to be elucidated. PMID- 12650969 TI - Changes in phosphorylation of ERK and Fos expression in dorsal horn neurons following noxious stimulation in a rat model of neuritis of the nerve root. AB - Mechanical compression and chemical inflammation of the spinal nerve root are considered major sensory pathologies secondary to a lumbar disc herniation. In order to elucidate the dorsal horn responsiveness to noxious stimulation to the peripheral tissue in the neuritis model of the nerve root, we examined extracellular signal-regulated kinase (ERK) phosphorylation and Fos expression in spinal cord dorsal horn neurons. Male Sprague-Dawley rats received hemilaminectomies and the implantation of disc tissue that was obtained from coccygeal intervertebral discs. Three or 7 days after surgery, rats were perfused after receiving noxious mechanical stimulation of the plantar surface of the hind paw using a hemoclip, and the L4/5 spinal cord was processed for immunohistochemistry with antibodies for phospho-ERK and Fos. The number of Fos immunoreactive (Fos-LI) neurons and phospho-ERK-immunoreactive (phospho-ERK-LI) neurons in the neuritis group after the noxious stimulation significantly increased compared to the sham-treated group at 3 and 7 days after surgery. The change in number of phospho-ERK-LI and Fos-LI neurons occurred mainly in the superficial dorsal horn. The number of Fos-LI neurons observed when the MEK inhibitor, U0126, was administered was significantly suppressed compared to the DMSO- (vehicle control) administered group. The increase in ERK phosphorylation and Fos expression in the spinal cord dorsal horn neurons indicates that responses/activation by the noxious stimulation applied to the periphery were elevated in spinal cord neurons in this neuritis model of the lumbar nerve root. Moreover, the increase in the Fos expression in the spinal cord dorsal horn may have been the result of the activation of the MAP kinase cascade. PMID- 12650970 TI - Characterization of wild-type (R100R) and mutated (Q100Q) GABAA alpha 6 subunit in Sardinian alcohol non-preferring rats (sNP). AB - Sardinian alcohol non-preferring (sNP) rats, selected for their low ethanol preference and consumption, carry a point mutation (R100Q) in the gene coding for GABA(A) receptor alpha(6) subunit, which becomes more sensitive to diazepam evoked GABA currents. We performed binding studies in the cerebellum of normal (RR) and mutated (QQ) sNP rats using [3H]Ro 15-4513, an inverse agonist for the benzodiazepine site which binds both diazepam insensitive and diazepam sensitive sites. Saturation curves performed on cerebellar membrane from genotyped rats indicated an higher affinity of [3H]Ro 15-4513 for GABA(A) receptors in QQ with respect to RR rats (K(d) values 4.0+/-0.67 and 6.24+/-0.95 nM, respectively), with similar B(max) values (3.5+/-0.25 and 3.9+/-0.39 pmol/mg protein, respectively). Diazepam displacement curves showed a two component model for both genotypes, with similar K(i1) values for QQ and RR (3.6+/-0.62 and 4.9+/-0.33 nM, respectively). In QQ rats diazepam is able to completely displace [3H]Ro 15-4513 (K(i2)=1.48+/-0.27 microM), while in RR rats the diazepam sensitive sites are still present (K(i2)>10 microM). The basal mRNA and protein expression level of the alpha(6) subunit were similar in RR and QQ rats. The electrophysiological profile of oocytes of Xenopus laevis injected with cerebellar synaptosomes showed that ethanol positively modulated GABA-evoked currents significantly more in QQ than in RR rats. These data contribute to the characterization of the function of GABA(A) alpha(6) subunit and its involvement in determining alcohol related behavior. PMID- 12650971 TI - Further evidence for the interaction of mu- and delta-opioid receptors in the antinociceptive effects of the dual inhibitor of enkephalin catabolism, RB101(S). A spinal c-Fos protein study in the rat under carrageenin inflammation. AB - We have previously shown that RB101, a dual inhibitor of enkephalin-degrading enzymes, decreased carrageenin-evoked c-Fos protein expression at the spinal cord level in awake rats. Moreover, we have also shown that c-Fos expression is a useful marker of the possible direct or indirect interactions between neural pathways, such as opioid and cholecystokinin systems. We now investigated the respective roles of the three main types of opioid receptors (mu, delta, or kappa) and their possible interactions, in the depressive effects of RB101 in inflammatory nociceptive conditions induced by intraplantar carrageenin (6 mg/150 microl of saline). We used beta-funaltrexamine (beta-FNA), naltrindole (NTI), and nor-binaltorphimine (BNI) as specific antagonists for mu, delta- and kappa-opioid receptors, respectively. c-Fos protein-immunoreactivity (c-Fos-IR) was evaluated as the number of c-Fos-IR nuclei in the lumbar spinal cord 90 min after carrageenin. c-Fos-IR nuclei were preferentially located in the superficial (I II) and deep (V-VI) laminae of segments L4-L5 (areas containing numerous neurons responding exclusively, or not, to nociceptive stimuli). RB101(S) (30 mg/kg, i.v.) significantly reduced the total number of carrageenin-evoked c-Fos-IR nuclei (30% reduction, P<0.01). This effect was completely blocked by beta-FNA (10 mg/kg, i.v.), or NTI (1 mg/kg, i.v.). In contrast, BNI (2.5 mg/kg, i.v.) did not reverse the reducing effects of RB101(S) on carrageenin-evoked c-Fos protein expression. These results suggest that functional interactions occur between mu- and delta-opioid receptors in enkephalin-induced antinociceptive effects. PMID- 12650972 TI - Identification of the requisite brain sites in the neuronal network subserving generalized clonic audiogenic seizures. AB - Comparative studies of neuronal networks that subserve convulsions in closely related epilepsy models are revealing instructive data about the pathophysiological mechanisms that govern these networks. Studies of audiogenic seizures (AGS) in genetically epilepsy-prone rats (GEPRs) and related forms of AGS demonstrate important network similarities and differences. Two substrains of GEPRs exist, GEPR-9s, exhibiting tonic AGS, and GEPR-3s, exhibiting clonic AGS. The neuronal network for tonic AGS resides exclusively in brainstem nuclei, but forebrain sites, including the amygdala (AMG), are recruited after repetitive AGS induction. The neuronal network for clonic AGS remains to be investigated. The present study examined the neuronal network for clonic AGS in GEPR-3s by microinjecting a competitive NMDA receptor antagonist, D,L-2-amino-7 phosphonoheptanoic acid (AP7), into the central nucleus of inferior colliculus (ICc), deep layers of superior colliculus (DLSC), periaqueductal grey (PAG), or caudal pontine reticular formation (cPRF), which are implicated in tonic AGS networks. Microinjections into AMG and perirhinal cortex (PRh), which are not implicated in AGS, were also done. AGS in GEPR-3s were blocked reversibly after microinjections into ICc, DLSC, PAG or cPRF. However, AGS were also blocked by AP7 in AMG but not PRh. The sites in which AP7 blocks AGS are implicated as requisite components of the clonic AGS network, and these data support a critical role for NMDA receptors in clonic AGS modulation. The brainstem nuclei of the clonic AGS network are identical to those subserving tonic AGS. However, the requisite involvement of AMG in the clonic AGS network, which is not seen in tonic AGS, is surprising and suggests important mechanistic differences between clonic and tonic forms of AGS. PMID- 12650973 TI - Identification of neurons containing orexin-B (hypocretin-2) immunoreactivity in limbic structures. AB - Orexins (hypocretins) are neuropeptides which have recently been identified exclusively within lateral hypothalamic and perifornical neurons, and these orexin (ox) containing neurons appear to have extensive projections to all levels of the neuraxis. In this study, we report the identification of two distinct clusters of neurons containing ox-B-like immunoreactivity within the amygdaloid complex of the rat. A cluster of small to medium size ovoid shaped neurons containing ox-B-like immunoreactivity was found predominantly within the lateral division of the central nucleus of the amygdala (ACe). A second distinct, but smaller group of ox-B labelled neurons with similar shapes and sizes to those in ACe was also identified in the anterior lateral subnucleus of the bed nucleus of the stria terminalis (BST) immediately adjacent the internal capsule, and in an area just ventral to the lateral ventricle. Neurons containing ox-A-like immunoreactivity were not observed in either structure. However, both structures contained ox-A- and ox-B labelled varicose fibers. Unilateral electrolytic lesions of the lateral hypothalamic area that contained ox-A and ox-B neurons did not alter the labelling of either ACe or BST ox-B pericarya. As both the ACe and BST are known to be involved in integrating complex homeostatic mechanisms associated with behaviours, these data suggest that a specific subset of ox-B neurons within the amygdaloid complex may serve as a component of neuronal circuits coordinating these responses. PMID- 12650974 TI - Differential expression of matrix metalloproteinases in monkey eyes with experimental glaucoma or optic nerve transection. AB - Extracellular matrix (ECM) remodeling after neuronal injury and reactive gliosis is carried out by activation of matrix metalloproteinases (MMPs) regulated by their tissue inhibitors (TIMPs). In glaucoma, there is a loss of retinal ganglion cells and extensive ECM remodeling (cupping) at the level of the optic nerve head, frequently associated with elevated intraocular pressure. To determine whether ECM remodeling in the glaucomatous optic nerve head occurs in response to loss of axons or to elevated intraocular pressure we compared the patterns of MMP and TIMP expression in the eyes of monkeys with laser-induced glaucoma or with optic nerve transection. MT1-MMP and MMP1 expression was markedly increased in reactive astrocytes in optic nerve heads with experimental glaucoma but not in the optic nerve head of transected eyes. In normal control eyes retinal ganglion cells expressed MMP2, TIMP1 and TIMP2 constitutively, and the proteins were detected in their axons. At the site of transection, MT1-MMP, MMP1, MMP2, TIMP1 and TIMP2 were expressed by reactive astrocytes. Inflammatory cells, fibroblasts and reactive astrocytes at the transected site expressed MMP3 and MMP9, which were undetectable in the retina and optic nerve head in any condition. Constitutive expression of MMP2, TIMP1 and TIMP2 in retinal ganglion cells suggests a role in maintenance of synaptic integrity and plasticity and maintenance of the periaxonal space. Increased MMP1 and MT1-MMP1 expression in the glaucomatous optic nerve head is specific to tissue remodeling due to elevated intraocular pressure and not secondary to loss of axons. PMID- 12650975 TI - Hippocampal synaptic plasticity in mice carrying the rd mutation in the gene encoding cGMP phosphodiesterase type 6 (PDE6). AB - Cyclic GMP (cGMP) has been implicated in the modulation of long-term potentiation (LTP) and depression (LTD) in the hippocampus. Transcripts for subunits of several types of cGMP specific phosphodiesterase are found in the mammalian brain but their relative role in hippocampal function is unclear. The retinal degeneration (rd) mutation in the gene encoding the PDE6B subunit causes a loss of function in PDE6 enzyme and in adult mice homozygous to the mutation it causes blindness. We have used this natural mutation, and the cGMP phosphodiesterase inhibitor zaprinast, in wild-type and rd/rd mouse littermates to investigate whether PDE5 and/or PDE6 regulates excitatory synaptic transmission in the hippocampus. Mice were genotyped using two independent PCR methods. Glutamate mediated synaptic transmission in the CA1 region or dentate gyrus was unaffected in hippocampal brain slices from mice carrying the rd mutation. Similarly the facilitation of synaptic events by paired-pulse stimuli, and LTP induced by a theta-burst (10 bursts of four events at 100 Hz with a 200-ms inter-burst interval) were normal in rd/rd mice. Inhibition of cGMP-specific PDE activity by zaprinast (10 microM, an inhibitor of PDE5 and PDE6) induced a slowly developing and sustained depression of field synaptic potentials that was quantitatively similar in both wild-type and rd/rd mice. Thus in the CA1 region synaptic plasticity is likely to be regulated by the PDE5 rather than the PDE6 isoform. PMID- 12650976 TI - Aberrant expression of peroxiredoxin subtypes in neurodegenerative disorders. AB - An increasing body of evidence indicates that oxidative stress and damage play a role in the pathogenesis of a number of diseases associated with neurodegeneration, including Down syndrome (DS), Alzheimer's disease (AD) and Pick's disease (PD). Although oxidative stress is a common element in these diseases, specific clinico-pathological phenotypes have been described for each disorder. Development of these phenotypes might be linked, among others, to differences in antioxidant response. The present study is designed to investigate expression of peroxiredoxins (Prxs), the newly characterized family of highly conserved antioxidant enzymes, and other antioxidant enzymes in frontal cortex and cerebellum of DS, AD and PD patients using the technique of proteomics. Levels of Prx I, Mn superoxide dismutase (SOD2) and glutathione-S-transferase omega1 in DS, AD and PD were not significantly different from that of controls in both brain regions investigated. In contrast, Prx II was significantly increased (P<0.05) in frontal cortex of DS, AD and PD, whereas Prx III was decreased in frontal cortex of DS (P<0.01) and PD (P<0.001). Interestingly, Prx VI displayed a significant increase (P<0.05) only in PD frontal cortex. The present data indicate that differential regulation of antioxidant enzymes exist in DS, AD and PD, suggestive of the diversity as well as distinct functional roles of these proteins. Moreover, while up-regulation of Prx II appears to provide evidence for the existence of compensatory response in increased cell loss, up-regulation of Prx VI may be used to discriminate PD from AD as well as DS. PMID- 12650977 TI - Autophosphorylated calcium/calmodulin-dependent protein kinase II alpha (CaMKII alpha) reversibly targets to and phosphorylates N-methyl-D-aspartate receptor subunit 2B (NR2B) in cerebral ischemia and reperfusion in hippocampus of rats. AB - It has been reported that cerebral ischemia induces Thr286 autophosphorylation and translocation of CaMKIIalpha which targets to and phosphorylates NR2B in hippocampus of rats [Neuroscience 96 (2000) 665; J. Biol. Chem. 275 (2000) 23798]. To further illustrate the mechanisms underlying these processes, we examined the effects of ketamine (a selective antagonist of NMDA receptor), KN-62 (1-[N,O-bis-(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine, a selective inhibitor of CaMKII) and reperfusion on CaMKII and NMDA receptors and the interactions between these signal proteins. Firstly, our results showed that ketamine decreased the ischemia-induced autophosphorylation, translocation and the targeting of CaMKIIalpha to NR2B and the serine-phosphorylation of NR2B. Secondly, KN-62 also inhibited the autophosphorylation of CaMKIIalpha, NR2B serine-phosphorylation and the binding of CaMKIIalpha to NR2B but had no effect on the translocation of CaMKII. These data strongly suggest that NMDA receptor channels mediated the Ca(2+)-dependent activation of CaMKII and NMDA receptors surely were the substrates on membranes of active CaMKII. Thirdly, our results indicated the concomitant phosphorylation and dephosphorylation of CaMKII and NR2B following ischemia or longer reperfusion. Moreover, the dissociation of CaMKII from NR2B had the same trend as that of the return of CaMKII to cytosol. All these data imply the close relationships between CaMKII and NR2B during ischemia and reperfusion, namely, CaMKII might act as an amplifier of detrimental cellular calcium signal regulated by NMDA receptors when becoming autophosphorylated and targeting to NR2B; conversely, autophosphorylated CaMKII could modulate NMDA receptor channel properties by phosphorylating NR2B. PMID- 12650978 TI - Modulation of DNA binding of nuclear transcription factors with leucine-zipper motifs by particular endogenous polyamines in murine central and peripheral excitable tissues. AB - Transcriptional regulation is one of the most important functions of polyamines in the nucleus of eukaryotic cells. The addition of the endogenous polyamines spermine and spermidine markedly increased DNA binding activity of the transcription factor activator protein-1 (AP1) in a concentration-dependent manner at a concentration range of 50 to 500 microM in nuclear extracts of murine whole brain when determined in the absence of added MgCl(2) on gel retardation electrophoresis. Similar but less potent potentiation was seen with DNA binding of cAMP responsive element binding protein (CREB), while both polyamines were ineffective in affecting that of c-Myc irrespective of the addition of MgCl(2). Unlabeled AP1 probe was invariably more potent in competing for AP1 binding than unlabeled CREB probe in either the presence or absence of spermine and spermidine. In addition to whole brain, both polyamines significantly increased AP1 binding in retina, adrenal and pituitary, without significantly affecting that in spleen. Moreover, ultraviolet and circular dichroism spectra analyses revealed that these two polyamines induced DNA topological transition of AP1 probe under the conditions favorable for the increase in AP1 binding. These results suggest that both spermine and spermidine may modulate gene transcription through cis- and trans-actions on AP1 binding in the nucleus in murine central and peripheral structures with high excitability. PMID- 12650979 TI - Baseline and 8-OH-DPAT-induced release of acetylcholine in the hippocampus of aged rats with different levels of cognitive dysfunction. AB - During aging, neurotransmission systems such as the cholinergic and serotonergic ones are altered. Using rats aged 3 or 24-26 months, this study investigated whether the well-described 8-OH-DPAT-induced increase of hippocampal acetylcholine release was altered in aged rats and whether it may vary according to the magnitude of age-related cognitive deficits. Long-Evans female rats aged 24-26 months were classified as good or bad performers on the basis of their reference-memory performance in a Morris water-maze task. Subsequently, the efficiency of 5-HT(1A) receptor agonist 8-OH-DPAT (0.5 mg/kg, s.c.) in triggering hippocampal acetylcholine release was evaluated by in vivo microdialysis and high performance liquid chromatography analysis. Besides a reduced baseline release in aged rats and a correlation between the baseline release and probe-trial performance in all rats, the results demonstrated that 8-OH-DPAT produced a significant increase of hippocampal acetylcholine release (peak value) in all rats, whether aged or young. While significant in bad performers (+56%), this increase did not reach significance in good performers (+32%). The results suggest that (i) some aspects of cognitive alterations related to aging might be linked to the baseline release of acetylcholine in the hippocampus, and (ii) the cholinergic innervation of the hippocampus of aged rats responds almost normally to systemic activation of 5-HT(1A) receptors, and (iii) differential alterations of cholinergic/serotonergic interactions assessed by determination of the 8-OH DPAT-induced release of acetylcholine in the hippocampus could not be linked with clarity to the cognitive status of aged rats. PMID- 12650980 TI - Alterations in D1/D2 synergism may account for enhanced stereotypy and reduced climbing in mice lacking dopamine D2L receptor. AB - Concurrent activation of dopamine D1 and D2 receptors (D1 and D2) is required for the expression of certain dopamine (DA)-mediated responses, such as climbing and stereotyped behaviors. Such interactions between D1 and D2 (i.e. D1/D2 synergism) represent an important aspect of dopaminergic function and plasticity. The D2 receptor exists in two isoforms: D2L and D2S. We have generated mice that selectively lack D2L (D2L-/-). Here we showed that treatment with the indirect DA agonist amphetamine, the direct DA agonist apomorphine, or combination of D1 and D2 agonists elicited intense climbing in wild type mice (which express predominantly D2L in the striatum), but this behavior was absent or reduced in D2L-/- mice. On the other hand, apomorphine, the D2 agonist quinpirole, or combination of quinpirole and the D1 agonist SKF 81297 induced more stereotyped behavior such as biting or head movements in D2L-/- mice (which express only D2S) than in wild type mice. The D1 receptor functioned normally in D2L-/- mice. Taken together, these results suggest that D2L and D1 interactions may play a greater role in DA agonist-induced climbing, whereas D2S and D1 interactions may have a larger impact on DA agonist-induced stereotypy (and possibly psychosis). DA agonists, which are clinically used to treat Parkinson's disease and attention deficit hyperactivity disorder (ADHD), are known to induce psychotic side effects. Thus, our findings may provide novel insights for designing anti parkinsonian, anti-ADHD and antipsychotic drugs with greater therapeutic efficacy and fewer side effects. PMID- 12650981 TI - The control of [125I]BDNF release from striatal rat brain slices. AB - The depolarisation-induced release of brain-derived neurotrophic factor (BDNF) from adult rat striatal slices was studied in vitro. The slices were preloaded with [125I]BDNF and exposed to depolarising stimulation with varying concentrations of veratrine (up to 50 microM) and potassium (up to 50 mM) which caused activity-dependent short-term release of [125I]BDNF. The results indicate that this stimulated release of [125I]BDNF is not regulated by a feedback mechanism mediated via the TrkB receptor. The release of [125I]BDNF was found to be dependent on the concentrations of both extracellular and intracellular calcium, since BDNF release was modulated by the addition of both EGTA and BAPTA AM, agents chelating either external or internal Ca(2+), respectively. BDNF release also proved to be dependent on activation of IP(3) mediated Ca(2+) release from intracellular stores. [125I]BDNF release was also modulated by 5HT(3) receptor ligands and by receptors coupled to adenylate cyclase. Taken together, these results indicate that [125I]BDNF release is activity dependent, and is modulated by changes in Ca(2+) levels. Moreover the release occurs via a mechanism involving cAMP. PMID- 12650983 TI - Immunohistochemical study on the distribution of homocysteine in the central nervous system of transgenic mice expressing a human Cu/Zn SOD mutation. AB - In the present study, we used the transgenic mice expressing a human Cu/Zn SOD mutation (SOD1(G93A)) as an in vivo model of ALS and performed immunohistochemical studies to investigate the changes of homocysteine in the central nervous system of symptomatic transgenic mice. In control and presymptomatic transgenic mice, homocysteine-immunoreactive astrocytes were not detected in any region. In symptomatic transgenic mice, homocysteine immunoreactive astrocytes were distributed in the spinal cord, brainstem and cerebellar nuclei of transgenic mice. In the hippocampal formation of transgenic mice, pyramidal cells in the CA1-3 regions and granule cells in the dentate gyrus showed homocysteine immunoreactivity. The present study provides the first in vivo evidence that homocysteine immunoreactive astrocytes were found in the central nervous system of symptomatic SOD(G93A) transgenic mice, suggesting that reactive astrocytes may play an important role in the pathogenesis and progress of ALS. This study also suggests that increased expression of homocysteine in the hippocampal neurons might reflect a role of homocysteine in an abnormality of hippocampal function of ALS. PMID- 12650982 TI - Huntingtin-interacting protein-1-related protein of rat (rHIP1R) is localized in the postsynaptic regions. AB - We cloned a rHIP1R (GenBank Accession No., AB005052) encoding a Sla2/huntingtin interacting protein (HIP1) family protein from a rat brain cDNA library. Localization of rHIP1R was investigated in the rat brain using an antibody specific to the HIP1R antibody. The rHIP1R protein was enriched in the synaptic plasma membrane fraction along with huntingtin, a synaptic protein and a causal protein for Huntington's disease. The electron microscopic examination revealed that HIP1R was localized at postsynaptic spines. Localization of HIP1R in the small vesicular structures in the spine, possible sites of vesicular transport of synaptic proteins, together with the structure-based analysis, suggested a role of HIP1R for vesicle trafficking through interaction with F-actin and working together with huntingtin and HIP1 at the synaptic sites. PMID- 12650984 TI - Differential modulation of light-evoked on- and off-EPSCs by paired-pulse stimulation in salamander retinal ganglion cells. AB - Short-term plasticity of On- and Off-EPSPs, and its potential role in regulation of signal processing was studied in salamander retinal On-Off ganglion cells by whole-cell recording. Paired-pulse light stimulation resulted in a depression of On-, and an enhancement of Off-EPSCs. Recovery from depression and enhancement was exponential and complete by 20 s. Paired-pulse enhancement, but not depression, was abolished with increasing stimulus duration. Blockade of On-EPSC by L-2-amino-4-phosphonobutyrate (AP-4), an agonist at group III mGluRs, significantly increased Off-EPSCs evoked by short (<2 s) duration conditioning light stimuli, resulting in a reversal of the paired-pulse enhancement to depression. The acetylcholinesterase inhibitor eserine reduced Off-EPSC1 and increased the ratio of enhancement. An opposite effect was observed in the presence of the nACh receptor antagonist d-tubocurarine. AP-7, an antagonist of NMDA receptors attenuated the enhancement of Off-EPSCs. In current clamp mode paired-pulse stimulation resulted in a modulation of light evoked, as well as the depolarization-induced spike firing pattern of ganglion cells. The present study suggests that paired light stimulation differently modulates On and Off EPSPs, and the light-evoked spike firing pattern of On-Off ganglion cells. PMID- 12650985 TI - A comparison of voluntary salt-intake behavior in Nax-gene deficient and wild type mice with reference to peripheral taste inputs. AB - The Na(x) channel, a subfamily of voltage-gated sodium channels, is thought to be a specific sodium receptor in the central nervous system. Our previous study revealed that Na(x)-gene-deficient mice consumed excessive amounts of NaCl even under water-deprived conditions. In the present study, to investigate whether the peripheral taste inputs are involved in the abnormal intake of salt in Na(x) deficient mice (homo), voluntary intake of various taste solutions in homo and wild-type mice (wild) was examined under non-deprived conditions. Homo showed a higher preference for 0.15 M NaCl solution than wild. Preference ratios for other basic tastants were identical between groups. Transection of the chorda tympani (CT) or the glossopharyngeal (GP) nerve had little effect on salt-intake behavior in homo and wild. Although combined transection of the superior laryngeal (SL) and GP nerves decreased NaCl intake in homo but not in wild, there were no differences in preference ratios for NaCl in homo before and after SL+GP transection. On the other hand, preference ratios for NaCl in wild tended to increase after combined SL and GP transection. Consequently, preference ratios for NaCl after SL+GP transection were no different between homo and wild. While electrophysiological responses of the CT and the GP to various taste solutions were indistinguishable between homo and wild, those of the SL to NaCl in homo were smaller than those in wild only at lower concentrations (0.01 and 0.03 M). Thus, chemosensory inputs from the oro-pharyngeal regions had little effect on abnormal salt intake in homo, if any. From these results, it is suggested that the higher preference for NaCl in homo is mainly due to the lack of Na(x) channels in the central nervous system. PMID- 12650986 TI - Topiramate reduces excitotoxic and ischemic injury in the rat retina. AB - The effects of topiramate, a drug used clinically as an anti-epileptic, were investigated in excitotoxin-induced neurotoxicity models involving two different retinal primary cultures and in a rat model of retinal ischemic injury. For the in vitro studies, we used retinal-neuron cultures from rat embryos and purified retinal ganglion cells (RGCs) from newborn rats. In the retinal-neuron cultures, neurotoxicity was induced by a 10-min exposure to 1 mM glutamate or (+/-)-a-amino 3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA). In RGCs, neurotoxicity was induced by incubation for 3 days in a culture medium containing 25 microM glutamate. For the in vivo study, retinal ischemia was induced by elevating intraocular pressure to 130 mmHg for 45 min, and topiramate was administered intraperitoneally before and after the ischemia. Retinal damage was evaluated by measuring the number of cells in the ganglion cell layer (GCL) and the thickness of the inner plexiform layer (IPL), and by examining the a- and b-waves of the electroretinogram (ERG). Topiramate (> or =1 microM) markedly reduced the neuronal cell death induced by each of the excitotoxins in rat retinal-neuron cultures and in RGCs. Ischemia caused a decrease in GCL cells and in IPL thickness, and a diminution of the ERG waves. Histopathologic and functional analyses indicated that systemic treatment with topiramate prevented ischemia induced damage in a dose-dependent manner. In conclusion, topiramate was protective against excitotoxic and ischemic retinal-neuron damage in vitro and in vivo, respectively. Therefore, it may be useful for treatment of the retina related diseases such as central retinal artery occlusion, diabetic retinopathy, and glaucoma. PMID- 12650987 TI - Consistency in the number of dopaminergic paraventricular organ-accompanying neurons in the posterior tuberculum of the zebrafish brain. AB - The teleostean diencephalon contains a relatively large number of dopaminergic neurons compared to other vertebrates. In the zebrafish, 17 groups of such neurons have been distinguished. One of the most unusual among these is the group of paraventricular organ-accompanying cells, which are easily distinguished by their large somal size, high tyrosine hydroxylase content, and characteristic dendritic architecture. This cell group is also heterogeneous-subsets of neurons can be identified on the bases of dendritic orientation. In this study, the number of paraventricular organ-accompanying neurons is counted in adult brain sections stained with anti-tyrosine hydroxylase antibodies. There is an average of 7.2+/-1.0 neurons on each side of the brain, and an average sum of 14+/-1.1 neurons on both sides. The average difference between the left and the right sides is 0.8+/-0.5 neurons. Neuron numbers between the two sides of the same brain are highly correlated. These results suggest that there is a relatively stringent regulation of paraventricular organ-accompanying neuron number in the zebrafish brain. The correlated left-right numbers suggests that genetic factors may play a major role in this regulation. The consistent and low cell number should be helpful in elucidating the number of subsets of these neurons, the anatomical and functional organization of some of the dopaminergic neurons along the paraventricular organ, as well as factors that play a role in regulating neuron numbers. PMID- 12650988 TI - Effect of acute ethanol administration and acute allopregnanolone administration on spontaneous hippocampal pyramidal cell neural activity. AB - We investigated the effect of acute ethanol administration and acute allopregnanolone administration on spontaneous hippocampal pyramidal cell neural activity. Both agents produced significant reductions in spontaneous firing rate of hippocampal pyramidal neurons at a medium and high doses. Furthermore, blockade of allopregnanolone biosynthesis by preadministration of finasteride, a 5alpha-reductase blocker, prevented ethanol-induced inhibition on hippocampal pyramidal neural activity. The results further demonstrate similar effects of allopregnanolone and ethanol on hippocampal neurophysiology and that allopregnanolone plays a key role in producing ethanol-induced inhibition of hippocampal neural activity. PMID- 12650989 TI - Localization of angiotensin II (AT1)-receptor-immunoreactive fibres in the hypothalamus of rats: angiotensin II-sensitive tanycytes in the ependyma of the third ventricle? AB - Scanning the hypothalamus of rats for receptor binding sites of the octapeptide hormone angiotensin II (ANG II), we observed ANG II-sensitive fibres in the ventrolateral hypothalamus. The ANG II (AT(1))-receptor-immunoreactive processes originate from cells-probably tanycytes-embedded in the base and the ventrolateral walls of the third ventricle and reach into the retrochiasmatic area, the ventrolateral hypothalamus and the median eminence. PMID- 12650990 TI - GABAA receptor epsilon subunit expression in identified peptidergic neurons of the rat hypothalamus. AB - Dual-labeling immunohistochemical or in situ hybridization studies for the recently cloned epsilon-subunit and several neuropeptides were performed in the rat hypothalamus. We revealed an extensive co-expression (>90%) with hypocretin (Hcrt), oxytocin (OT), the gonadotropin-releasing hormone (GnRH), and the melanin concentrating hormone (MCH) peptides, whereas occasional co-expression (<10%) with cocaine-amphetamine-regulated transcript (CART) was found. Our results suggest that novel GABA(A) receptor subtypes comprising epsilon-subunit are important for metabolic and neuroendocrine functions. PMID- 12650991 TI - Effects of the cannabinoid antagonists AM281 and AM630 on deprivation-induced intake in Lewis rats. AB - Male Lewis rats (two groups of 10) received intracerebroventricular injections of either AM 630 (vehicle, 2.5, 5, 10 and 20 microg) or AM 281 (vehicle, 5, 10, 20 and 40 microg) following overnight food deprivation. The CB2 antagonist AM 630 failed to block deprivation-induced intake at 0.5, 1, 2, 4 and 6 h. The CB1 antagonist AM 281 significantly blocked intake following 20 microg (1 h) and 40 microg (1, 2, 4 and 6 h). Results are discussed with respect to cannabinoid receptor systems' involvement in ingestion and the differential pharmacological profiles of AM 630 and AM 281. PMID- 12650992 TI - Degenerative patterns following denervation of temporal structures in a rat model of mnemonic dysfunction. AB - The temporal region makes up an important substrate for mnemonic function in both humans and animals. Rats with transection of fibers connecting the temporal and entorhinal cortices display impaired visual memory. The use of hematoxylin and eosin (HE) or Nissl staining has proved insufficient for characterization of the degenerative events that take place after the lesions, and in the present study we therefore applied Fluoro-Jade dye, and zinc autometallography (AMG) in order to describe degenerative changes with AMG. We found that 14 days survival resulted in a much more severe degeneration of cell bodies in the temporal, perirhinal, entorhinal, and postrhinal cortices than was seen after 8 days survival. Corresponding degeneration of cell bodies was optimally observed 7 days following surgery with the Fluoro-Jade staining. HE staining did not reveal degenerative changes with survival times of 8 or 14 days. A substantial increase in AMG staining for zinc ions after lesion revealed an increase in zinc enriched neuronal terminals in the temporal areas and was interpreted as sprouting from local zinc enriched neuronal projections. The present data are related to the memory deficits observed in rats with similar denervations in previous studies. PMID- 12650993 TI - Extracellularly applied horseradish peroxidase increases the number of dense core vesicles in leech sensory neurons. AB - The uptake of horseradish peroxidase (HRP), applied as an extracellular tracer, is a classical method for studying endo/exocytosis of synaptic vesicles at the ultrastructural level. It is generally not considered that HRP may affect neuronal function. Reported here is the finding that extracellularly applied HRP (0.1%) perturbs dense core vesicles in the synaptic processes of leech neurons. The strength of the effect varies with neuronal class. In sensory afferents, the number of dense core vesicles increases 5-fold, while there is only a 2-fold increase in central neurons. PMID- 12650994 TI - Exogenous NGF restores endogenous NGF distribution in the brain of the cognitively impaired aged rat. AB - Alzheimer's disease and normal aging may impair retrograde transport of nerve growth factor (NGF) from cortical areas to basal forebrain cholinergic neurons. We demonstrate a relationship between performance in a spatial reference memory task and NGF distribution in the aged rat brain. In addition, exogenous NGF restored endogenous NGF distribution in cognitively impaired aged rats. These data suggest that NGF administration restores utilization of endogenous growth factor in the brain of cognitively impaired aged rats. PMID- 12650996 TI - Purification of the proline-rich homeodomain protein. AB - The proline-rich homeodomain protein (PRH), also known as Hex, is a transcriptional repressor expressed in a variety of cell types. The PRH protein contains a proline-rich N-terminal domain that can repress transcription when attached to a heterologous DNA binding domain, a central homeodomain that mediates sequence-specific DNA binding, and an acidic C-terminal domain of unknown function. Although individual domains of PRH have been expressed in bacterial cells as GST- and histidine-tagged fusion proteins, attempts to express and purify the full-length protein have met with little success. Here we describe the purification of a histidine-tagged full-length PRH fusion protein. The protein described here will allow us to determine the mechanisms whereby PRH represses transcription. PMID- 12650997 TI - Purification of human alpha-L-fucosidase precursor expressed in Escherichia coli as a glutathione S-transferase fusion protein. AB - Alpha-L-fucosidase (FUC) is a glycosidase involved in the degradation of fucose containing glycoconjugates. A cDNA representing the complete sequence of human FUC was inserted into the prokaryotic expression vector pGEX-2T. High levels of the glutathione S-transferase (GST) fusion protein were detected in Escherichia coli cells after induction with isopropyl thio-beta-D-galactopyranoside. The GST FUC protein was mostly found as inclusion bodies and attempts to optimise its expression as a soluble form were unsuccessful. Nevertheless, the recombinant protein was purified by affinity chromatography on glutathione-sepharose and its fucosidase activity was characterised. After thrombin cleavage of the GST tag, the FUC precursor protein was purified by electro-elution. PMID- 12650998 TI - Mutation of surface-exposed histidine residues of recombinant human granulocyte colony stimulating factor (Cys17Ser) impacts on interaction with chelated metal ions and refolding in aqueous two-phase systems. AB - Site directed mutagenesis of Cys17-->Ser17 form of recombinant human granulocyte colony stimulating factor (rhG-CSF C17S) for sequential replacing of surface His(43) and His(52) with alanine was used to identify residues critical for the protein interaction with metal ions, in particular Ni(2+) chelated by dye Light Resistant Yellow 2 KT (LR Yellow 2KT)-polyethyleneglycol (PEG), and refolding after partitioning of inclusion bodies in aqueous two-phase systems. Strong binding of rhG-CSF (C17S) to PEG-LR Yellow 2KT-Cu(II) complex allowed for the adoption of affinity chromatography on Sepharose-LR Yellow 2KT-Cu(II) that appeared to be essential for the rapid isolation of mutated forms of rhG-CSF. Efficiency of that purification stage is exemplified by isolation of rhG-CSF (C17S, H43A) and rhG-CSF (C17S, H43A, H52A) mutants in correctly folded and highly purified state. Affinity partitioning of rhG-CSF histidine mutants was studied in aqueous two-phase systems containing Cu(II), Ni(II) and Hg(II) chelated by LR Yellow 2KT-PEG at pH 7.0 and Cu(II)-at pH 5.0. It was determined, that affinity of rhG-CSF mutants for metal ions decreased in the order of C17S>C17S, H43A>C17S, H43A, H52A for Cu(II), and C17S=C17S, H43A>C17S, H43A, H52A for Ni(II) ions, while affinity of all rhG-CSF mutants for Hg(II) ions was of the same order of magnitude. Influence of His(43) and His(52) mutation on protein refolding was studied by partitioning of the respective inclusion body extract in aqueous two-phase systems containing Ni(II) and Hg(II) ions. Data on rhG-CSF histidine mutant partitioning and refolding indicated, that His(52) mutation is crucial for the strength of protein interaction with chelated Ni(II) ions and refolding efficiency. PMID- 12650999 TI - Two-step purification of the outer membrane transporter and activator protein ShlB from Escherichia coli using internally His6-tagged constructs. AB - ShlB from Serratia marcescens was isolated and purified from a porin-deficient Escherichia coli BL21 strain using a combination of detergent extraction, affinity and ion-exchange chromatography. An internal histidine affinity tag was introduced that did not interfere with activity. At each stage of the purification scheme biological activity of the ShlB protein was assessed. Using this scheme, several His(6)-tagged mutants of ShlB were purified to electrophoretic homogeneity. PMID- 12651000 TI - Purification of recombinantly expressed human cluster determinant 4 cytoplasmic domain. AB - A DNA fragment coding for the human CD4 cytoplasmic domain (residues 394-433) was cloned into the pET15b expression vector. The resulting plasmid was used for synthesis of the polyhistidine-tagged 5.10(3) M(r) CD4 peptide in Escherichia coli BL21(DE3)Star. The CD4 cytoplasmic domain was purified under denaturing and reducing conditions by a two-step procedure using immobilized metal affinity chromatography and gel permeation chromatography. The purified CD4 cytoplasmic domain is soluble and functional without any specific refolding steps. The yield of the described purification procedure was approximately 5 mg peptide per liter culture volume. PMID- 12651001 TI - Purification of recombinant BtpA and Ycf3, proteins involved in membrane protein biogenesis in Synechocystis PCC 6803. AB - The gene products Ycf3 (hypothetical chloroplast open reading frame) and BtpA (biogenesis of thylakoid protein) are thought to be involved in the biogenesis of the membrane protein complex photosystem I (PSI) from Synechocystis PCC 6803. PSI consists of 12 different subunits and binds more than 100 cofactors, making it a model protein to study different aspects of membrane protein biogenesis. For a detailed biophysical characterization of Ycf3 and BtpA pure proteins must be available in sufficient quantities. Therefore we cloned the corresponding genes into expression vectors. To facilitate purification we created His-tagged versions of Ycf3 and BtpA in addition to the unmodified forms. Immobilized metal affinity chromatography (IMAC) yielded His-tagged proteins which were used for the production of antibodies. Purification strategies for non-tagged proteins could also be established: Ycf3 could be purified in soluble form using a two step purification in which ammonium sulfate precipitation was combined with anion exchange chromatography (IEC). BtpA had to be purified from inclusion bodies by two-consecutive IEC steps under denaturing conditions. An optimized refolding protocol was established that yielded pure BtpA. In all cases, MALDI-TOF peptide mass fingerprinting (PMF) was used to confirm protein identity. Initially, size exclusion chromatography and CD-spectroscopy were used for biophysical characterization of the proteins. Both Ycf3 and BtpA show homo-oligomerization in vitro. In summary, purification protocols for Ycf3 and BtpA have been designed that yield pure proteins which can be used to probe the molecular function of these proteins for membrane protein biogenesis. PMID- 12651002 TI - Expression and purification of Plasmodium falciparum MSP-1(42): A malaria vaccine candidate. AB - The C-terminal 42.10(3) Da portion of the merozoite surface protein (MSP-1) of the human malaria parasite Plasmodium falciparum is of interest, not only because it may constitute an essential part of a future anti-malaria vaccine, but also due to its role during the infection of erythrocytes by the parasite. We have cloned and expressed two synthetic DNA sequences encoding the two prototypic MSP 1(42) variants in E. coli. When over-produced, both proteins form insoluble aggregates which were isolated in high purity and yield. After solubilisation and refolding in vitro, both proteins were purified to homogeneity by a three-step procedure applying Ni-chelate, size exclusion and immuno-affinity chromatography. After purification, both proteins meet key criteria of preparations for clinical use. First, conformational studies suggest proper folding of the proteins, particularly in the region containing two EGF-like domains. Polyclonal serum raised against E. coli produced MSP-1(42) recognizes native MSP-1 in Plasmodium infected erythrocytes as shown by immunofluorescence. PMID- 12651003 TI - Cloning, purification and biochemical characterization of dipetarudin, a new chimeric thrombin inhibitor. AB - The development of thrombin inhibitors could provide invaluable progress for antithrombotic therapy. In this paper, we report the cloning, purification and biochemical characterization of dipetarudin, a chimeric thrombin inhibitor composed of the N-terminal head structure of dipetalogastin II, the strongest inhibitor from the assassin bug Dipetalogaster maximus, and the exosite 1 blocking segment of hirudin, connected through a five glycine linker. The cloning of dipetarudin was performed by a simple method which had not been used previously to clone chimeras. Biochemical characterization of dipetarudin revealed that it is a slow, tight-binding inhibitor with a molecular mass (M(r)=7560) and a thrombin inhibitory activity (K(i)=446 fM) comparable to r hirudin. PMID- 12651004 TI - Identification and characterisation of Schizosaccharomyces pombe cyclophilin 3, a cyclosporin A insensitive orthologue of human USA-CyP. AB - We have identified nine cyclophilins encoded in the genome of the fission yeast Schizosaccharomyces pombe (Sp). Cyclophilin 3 is an orthologue of hUSA-CyP, which is associated with Prp4/Prp3 in the [U4/U6.U5] snRNP complex and Prp18, both of which are components of the pre-mRNA splicing machinery. PPIase assays have shown SpCyp3 and hUSA-CyP to have comparable activity and substrate specificity, but SpCyp3 has a reduced sensitivity to CsA correlating with a difference in the catalytic site. Prp3, Prp4 and Prp18 proteins exist in S. pombe and nuclear localisation of SpCyp3 has been shown, indicating conservation of function between hUSA-CyP and SpCyp3. PMID- 12651005 TI - Novel approach to obtain biologically active recombinant heterodimeric proteins in Escherichia coli. AB - The strategy described in this paper provides a novel approach for recombinant expression of heterodimeric proteins, and is especially suitable for the production of proteins whose characteristics lead to aggregation in E. coli expression systems. Pheromaxein, a steroid-binding protein isolated from boar saliva, is a heterodimeric protein consisting of 10x10(3) rel. mol. mass units (pheromaxein A) and 8x10(3) rel. mol. mass units (pheromaxein C) subunits. Expression of pheromaxein subunits in E. coli resulted in extensive insoluble aggregation. The difficulty faced in obtaining soluble recombinant pheromaxein subunits was clearly evident when native pheromaxein immediately formed aggregates when it was separated into its individual subunits. An increase in soluble pheromaxein expression in E. coli was obtained when the subunits were expressed as fusion proteins with thioredoxin. Pheromaxein genes were inserted separately into pET32a+ vectors at the NcoI site, resulting in thioredoxin, S.Tag and His.Tag coding regions being upstream of the inserted gene. Soluble pheromaxein A-thioredoxin (pheroA/trx) and pheromaxein C-thioredoxin (pheroC/trx) fusions were purified to homogeneity, using a laboratory scale S-protein agarose affinity column. Cleavage of thioredoxin under normal conditions was not feasible due to the extensive aggregation problems experienced when pheromaxein subunits exist separately. PheroA/trx and pheroC/trx were therefore mixed together and cleaved from thioredoxin simultaneously so that pheromaxein subunits were given an instant opportunity to associate under oxido-shuffling conditions. The glutathione oxido-shuffling system allowed the disulphide bridges between pheromaxein A and C to rearrange until the correct native structure was formed. This novel approach combines affinity purification with a coupled fusion protein cleavage and refolding technique. PMID- 12651006 TI - Recombinant expression and purification of Ssa1p (Hsp70) from Saccharomyces cerevisiae using Pichia pastoris. AB - Heat shock proteins with a molecular mass of 70000 (Hsp70s) are a ubiquitous class of ATP-dependent molecular chaperones involved in the folding of cellular proteins. Sequencing the entire genome of Saccharomyces cerevisiae revealed 14 different genes for Hsp70 proteins in different cellular compartments. Among these 14 Hsp70s, the subclass of Ssa (Ssa1p-Ssa4p) is abundant and essential in the cytosol. Since high yield expression of cytoplasmic Ssa1p is inefficient in Saccharomyces cerevisiae and recombinant expression in E. coli yields low protein levels, we chose Pichia pastoris as the recombinant expression system. In Pichia pastoris, expression levels of Ssa1p are high and Ssa1p is soluble and correctly folded. Also, we present a new protocol for purification of Ssa1p. Previously described purifications include ATP-agarose chromatography leading to Ssa1p partially complexed with ATP. Our optimized purification protocol follows the CiPP strategy (capture, intermediate purification, polishing) avoiding ATP agarose chromatography, which allows detailed studies on the ATP-dependent Hsp70 functions. We obtained Ssa1p in high purity and 400 times higher quantity compared to previous studies. PMID- 12651007 TI - Cloning, expression and purification of three Chaperonin 60 homologues. AB - The Chaperonin 60 (Cpn60) proteins have, in addition to their well-known functions of protein folding and protection, a range of intercellular signalling activities. As part of a study to investigate the biological activity of the Cpn60 proteins, particularly from pathogenic organisms, we have cloned and expressed three Cpn60 proteins from Homo sapiens, Helicobacter pylori and Chlamydia pneumoniae. The Cpn60 proteins were purified to apparent homogeneity using a combination of nickel column affinity chromatography and Reactive Red dye affinity columns. Insoluble protein was solubilised using 8 M urea and then re folded on the nickel column by stepwise removal of the urea. The immunostimulant LPS was removed by addition of the antibiotic polymyxin B as part of the purification process. PMID- 12651008 TI - GroE-dependent expression and purification of pig heart mitochondrial citrate synthase in Escherichia coli. AB - Citrate synthase (CS) is a dimeric, mitochondrial protein, composed of two identical subunits (M(r) 48969 each). The nuclear-encoded alpha-helical protein is imported into mitochondria post-translationally where it catalyses the first step of the citric cycle. Furthermore, the pathway of thermal unfolding as well as the folding pathway was studied extensively, making CS a well-suited substrate protein for studying chaperone function. In chaperone research the quality of the substrate proteins is essential to guaranty the reproducibility of the results. In this context, we here describe the GroE-enhanced recombinant expression and purification of CS. CS was expressed in E. coli by using an arabinose regulated T7 promotor. Under standard expression conditions only insoluble, inactive CS was detected. Interestingly, the expression of soluble and active CS was possible when GroEL/GroES was co-expressed. Furthermore, a shift to lower expression temperatures increased the amount of soluble, active CS. We describe for the first time, the purification of CS in soluble and active form by following a CiPP strategy (capture, intermediate purification, polishing). After the initial capturing step on DEAE-Sephacel the protein was further purified on a Q-Sepharose column. After these two steps of anion-exchange chromatography a final size exclusion chromatography step on a Superdex 75-pg column yields CS with a purity over 99%. Using this expression and purification strategy 1 mg CS per g E. coli wet weight were purified. PMID- 12651009 TI - In vitro cell free synthesis of human manganese superoxide dismutase with the RTS 500 system. AB - Reduced activity of manganese superoxide dismutase (MnSOD) is the basis of several pathologic features and complications occurring in the course of infectious mononucleosis. In order to supply future research with easily accessible enzyme, an in vitro protocol was developed based on the RTS 500 system and an overexpression vector. Translation of MnSOD monomers could be detected by SDS-PAGE, and assembly of the active homotetramer by native PAGE. Enzyme activity was successfully shown by in gel activity tests and enzyme assays. With 15 micro g of DNA, 2.45 micro kat were generated. The purification of MnSOD was performed by chromatography applying the His-tag technology. In SDS-PAGE of the eluate, a band showed up at M(r) 25000. PMID- 12651010 TI - Simplified procedure to recover recombinant antigenized secretory IgA to be used as a vaccine vector. AB - Induced protection mechanisms at mucosal surfaces involve secretory IgA (SIgA), a complex structure made of polymeric-dimeric IgA (IgA(p/d)) antibody associated with secretory component (SC). SIgA can adhere to M cells of the intestinal and nasal epithelia, are transported across these latter, and are thus available to the immune cells underlying the epithelia. This property makes SIgA suitable as potential mucosal vaccine delivery vector. It remains that production and purification of SIgA is a complex task since IgA(p/d) and SC are naturally synthesized by two different cell types. Furthermore, only IgA(p/d) are capable to associate with SC. Thus, we sought to separate IgA(p/d) and monomeric IgA (IgA(m)) antibodies secreted by hybridoma cells in CELLine bioreactors. To this aim, we connected together two 1-m long columns filled with Sephacryl S-300 beads and placed them under the control of a automatized chromatographic system. In parallel, we produced recombinant antigenized human SC (ra-hSC) in Chinese hamster ovary (CHO) cells adapted to suspension culture in CELLine bioreactors. To avoid intermediate purification of ra-hSC, culture supernatants (SN) containing this latter were combined with purified IgA(p/d), and the recombinant antigenized SIgA (raSIgA) complex was resolved on a 1-m long column filled with Superdex 200 beads. Biochemical characterization based on SDS-PAGE, silver staining, immunodetection and enzyme-linked immunosorbent assay (ELISA) indicates that highly purified raSIgA can be recovered using this simple two-step procedure. Such preparations are currently used to immunize mice to induce mucosal and systemic responses. PMID- 12651011 TI - Cloning, expression and two-step purification of recombinant His-tag enhanced green fluorescent protein over-expressed in Escherichia coli. AB - In this report, we describe a two-step chromatographic procedure for the purification of His-tag EGFP by immobilized metal affinity expanded bed adsorption (IMAEBA) as the capture step and size exclusion chromatography as the polishing step. The use of proteins including a histidine-tag facilitates their subsequent purification after expression in many microorganisms. This meets the needs of scientific researchers as well as industrialists in purifying recombinant proteins. The procedure described allowed the obtention of 230 mg pure EGFP from 1 l simple batch culture with a recovery of 90%. PMID- 12651012 TI - Optimizing expression and purification from cell culture medium of trispecific recombinant antibody derivatives. AB - Antibody fragments offer the possibility to build multifunctional manifolds tailored to meet a large variety of needs. We optimized the production of a manifold consisting of one (bibody) or two (tribody) single-chain variable fragments coupled to the C-terminus of Fab chains. Different strong mammalian promoters were compared and the influence of expression media on production and recovery was investigated. Since the physical and chemical nature of these molecules largely depends on the nature of the antibody building blocks incorporated, a generally applicable process for the purification of recombinant antibody derivatives from serum containing mammalian cell culture medium was designed. To this end we compared protein L, hydroxyapatite, immobilized metal affinity chromatography, cation-exchange chromatography and hydrophobic charge induction chromatography. PMID- 12651013 TI - High-level expression and rapid purification of rare-codon genes from hyperthermophilic archaea by the GST gene fusion system. AB - In this study, we compared two gene fusion expression strategies using two rare codon genes (Ssh10b and MtGrxM) from archaea as a model system. Both genes can be highly expressed as N- or C-terminal fusion partners to GST or the intein/chitin binding tag. However, the fusion protein with intein tag could not be cleaved, even under stringent conditions, possibly due to steric hindrance, thus preventing further purification. In contrast, the GST fusion system could increase protein expression level and the corresponding fusion protein could be easily cleaved by thrombin. After binding to glutathione sepharose, the fusion protein was cleaved on column, and a roughly purified protein fraction was eluted. This fraction was purified by heating at 80 degrees C for 10 min, followed by centrifugation. The correct total mass and N-terminal primary structure were confirmed by mass spectrometry and Edman degradation. Both constructs were used for in vitro expression, and similar results were obtained, indicating higher expression levels of the GST tag vs. intein/chitin tag. Taken together, our results suggest that the GST fusion system can be used as a considerable alternative to synthetic genes for the expression of rare codon genes. The affinity chromatography purification followed by a heating step is an efficient and convenient method for thermostable protein purification. PMID- 12651014 TI - Optimisation of expression and purification of the recombinant Yol066 (Rib2) protein from Saccharomyces cerevisiae. AB - Yeast protein Yol066 (encoded by YOL066 ORF, also known as Rib2) possesses two distinct sequence domains: C-terminal deaminase domain and N-terminal part related to RNA:pseudouridine (psi)-synthases. The deaminase domain is implicated in the riboflavine biosynthesis, while the exact function of the RNA:Psi-synthase domain remains obscure. Here we report the optimisation of growth conditions and purification scheme for recombinant His(6)-tagged Yol066 expressed in E. coli BL21(DE3) using pET28 plasmid. Production of soluble Yol066 protein is best at low temperature (18 degrees C) and IPTG concentration (50 micro M) and Yol066 purification was achieved using metal-affinity and ion-exchange chromatography. This optimised protocol yields about 10 mg of highly purified recombinant Yol066 from 3 l of E. coli culture. PMID- 12651015 TI - Overexpression and purification of the three components of the Enterobacter aerogenes AcrA-AcrB-TolC multidrug efflux pump. AB - The tripartite AcrA-AcrB-TolC system is the major efflux pump of the nosocomial pathogen Enterobacter aerogenes. AcrA is a trimeric periplasmic lipoprotein anchored in the inner membrane, AcrB is an inner membrane transporter and TolC is a trimeric outer membrane channel. In order to reconstitute the AcrA-AcrB-TolC system of E. aerogenes in artificial membranes, we overexpressed and purified the three proteins. The E. aerogenes acrA, acrB and tolC open reading frames were individually inserted in the expression vector pET24a(+), in frame with a sequence coding a C-terminal hexahistidine tag to allow purification by INAC (Immobilized Nickel Affinity Chromatography). The mature AcrA-6His was overproduced in a soluble form in the cytoplasm of Escherichia coli BL21(DE3). AcrA-6His was purified under native conditions in two steps using INAC and gel permeation chromatography. We obtained about 25 mg of 97% pure AcrA-6His per liter of culture. AcrB-6His was solubilized from the membrane fraction of E. coli C43(DE3) in 300 mM NaCl, 5% Triton X-100 and purified in one step by INAC. The AcrB-6His enriched fraction was eluted with 100 mM imidazole. The final yield was 1-2 mg of 95% pure AcrB-6His per liter of culture. The membrane fraction of E. coli BL21(DE3)pLysS containing TolC-6His was first treated with 2% Triton X-100, 30 mM MgCl(2) to solubilize the inner membrane proteins. After ultracentrifugation, the pellet was treated with 5% Triton X-100, 5 mM EDTA to solubilize the outer membrane proteins. Approximately 5 mg of 95% pure TolC-6His trimers per liter of culture was purified by INAC. PMID- 12651016 TI - Production of recombinant thermostable proteins expressed in Escherichia coli: completion of protein synthesis is the bottleneck. AB - Heterologous expression and high yield purification of proteins are frequently required for structural and functional investigations. Purification of recombinant thermostable proteins is essentially trivial since unwanted mesophilic host protein can efficiently be removed by heat denaturation. However, heterologous expression in E. coli often results in truncated protein forms. In many cases, this is a consequence of abundant codons in heterologous genes, which are decoded by rare tRNAs in E. coli-a combination that can be responsible for translational stalling and termination during protein biosynthesis. Other complications may originate from potential initiation codons and ribosomal binding sites present inside the open reading frame of the target gene or from other less well defined phenomena such as mRNA instability. Separation of full length protein from truncated forms is a serious chromatographic problem that can be solved in the expression step. We have investigated the heterologous expression and purification of two translation initiation factors from the hyperthermophilic sulphate-reducing archaeon, Archaeoglobus fulgidus. Expression in E. coli was optimised to avoid truncated forms completely by complementation with the plasmids pSJS1244, pRIG, pCODON+ and pLysSR.A.R.E harbouring and expressing genes encoding rare tRNAs corresponding to the codons AGA, AGG, AUA, CUA, GGA, AAG and CCC. Two expression strains, C41(DE3) and C43(DE3) were found highly advantageous when combined with rare tRNA encoding plasmids as compared to BL21(DE3). We have also investigated the effects of site directed mutagenesis on rare lysine encoding AAG doublets as well as two methionine residues preceded by potential ribosomal binding sites. The expression approach presented here has enabled us to purify gram quantities of full-length protein by one step of ion exchange chromatography and is generally applicable to many other heterologously expressed thermostable proteins. PMID- 12651017 TI - Heterologous expression and purification of native and mutated low molecular mass glutenin subunits from durum wheat. AB - Wheat technological properties are correlated with the size of glutenin polymers, consisting of high and low molecular mass glutenin subunits, linked together by disulphide bonds. In order to unravel glutenin polymer structure, we considered three LMW-GS genes, which differ in the number of cysteine residues and in the repetitive domain length. The three LMW-GS genes have been expressed in Escherichia coli, and purified with a yield of 40-100 mg/l of culture volume, depending on protein type. Single polypeptides are being used in re-oxidation and micro-mixographic experiments, in order to detect the influence of the differential structural characteristics on glutenin polymer formation. PMID- 12651018 TI - Cloning and expression of fatty acids biosynthesis key enzymes from sunflower (Helianthus annuus L.) in Escherichia coli. AB - To further characterize the stearoyl-acyl carrier protein (ACP) desaturase (EC 1.14.99.6) and the acyl-ACP thioesterase FatB (EC 3.1.2.14) activities from sunflower seeds, we cloned, sequenced and expressed the recombinant genes in Escherichia coli. We obtained two partially purified proteins, His-SAD and His FATB, each of about 45000 Da. The expression of either proteins produced changes in the E. coli fatty acid profile indicating the functionality of the recombinant proteins. While the expression of His-SAD produced an effect similar to that produced by overexpression of the fabA gene, responsible for the fatty acid desaturation in E. coli, the expression of His-FATB gave rise to an unbalance between unsaturated fatty acids and a toxic effect in E. coli. PMID- 12651019 TI - Optimisation of expression and immobilized metal ion affinity chromatographic purification of recombinant (His)6-tagged cytochrome P450 hydroperoxide lyase in Escherichia coli. AB - Fatty acid hydroperoxide lyase (HPL) is a cytochrome P450 acting on fatty acid's hydroperoxides in many plants. The optimisation of the expression of recombinant (His)(6)-tagged HPL in Escherichia coli is described: the highest HPL production yield were obtained with TB medium supplemented with 2.5 mM delta-aminolevulinic acid and 0.5 mM IPTG. For the first time, the time course expression of a plant P450 in a bench-scale fermentor is detailed and the amount of recombinant HPL production is 16.3 mg/l. The UV-Visible spectrum of the recombinant (His)(6) tagged HPL have been recorded after a Ni(2+)-based affinity chromatography (IMAC). PMID- 12651020 TI - Continuous processing of fusion protein expressed as an Escherichia coli inclusion body. AB - In this study we develop the components of an integrated process for the continuous extraction and purification of a histidine-tagged fusion protein expressed as an inclusion body in Escherichia coli. Lac21 was selected as a model peptide and was expressed as a fusion to ketosteroid isomerase. A purification strategy was developed on a 1-ml batch column before successful scale-up and transfer to a continuous purification system, having a bed volume of 240 ml. Preliminary experiments proved cleavage of the fusion protein. The use of chemical extraction and continuous chromatography gives a flowsheet far superior to the traditional methods for inclusion body processing. PMID- 12651021 TI - A novel strategy for the purification of recombinantly expressed unstructured protein domains. AB - We recently found that the larger parts of the endocytic proteins epsin 1 and AP180 consist of an unstructured polypeptide chain. As a result these segments are completely heat-stable without loss of their functional properties. We have taken advantage of this fact and developed a combined heat lysis and pre purification procedure after expressing the disordered domains in E. coli. This results in the irreversible denaturation and precipitation of the majority of bacterial proteins. The bacteria are resuspended in a non-denaturing buffer, heated in a boiling water bath and shock-cooled. We demonstrate that this procedure compared to conventional lysis improves both yield and quality of the purified protein. PMID- 12651022 TI - Taenia solium cDNA sequence encoding a putative immunodiagnostic antigen for human cysticercosis. AB - A T. solium metacestode cDNA library was prepared and antibody screened to obtain recombinant antigens, which could be used for the neurocysticercosis diagnosis. The F18 clone was selected and sequenced, and the full length cDNA characterised as well as the genomic structure from the gene. F18 is a single copy gene that spans approximately 6.1 kb and contains five exons and four introns. The F18 cDNA has a 690-nucleotide open reading frame that encodes a putative polypeptide of 229 amino acids with a predicted molecular mass of 26.06 x 10(3) M(r). The F18 recombinant protein was obtained and purified by affinity chromatography using pGEX system (G-F18) and pQE system (H-F18). The purified G-F18 fusion protein showed the best results when it was used in ELISA with sera from neurocysticercosis patients. PMID- 12651023 TI - Large scale protein production of the extracellular domain of the transforming growth factor-beta type II receptor using the Pichia pastoris expression system. AB - To study the (patho)physiological role of transforming growth factor-beta (TGF beta), potent and selective inhibitors are necessary. Since TGF-beta signaling is initiated by the high affinity binding to the type II receptor (RII), the extracellular part of RII (solRII) can function as a TGF-beta antagonist. SolRII was cloned and large-scale protein synthesis was performed in the yeast Pichia pastoris expression system. Our results indicate that via this system, high levels of pure concentrated solRII can be obtained. Moreover, purified solRII is an active protein as shown by ELISA and bioassay. In conclusion, our large-scale protein expression procedure results in high quantities of purified solRII, which is a powerful tool to study the natural role of TGF-beta. PMID- 12651024 TI - Isolation of Qbeta polymerase complexes containing mutant species of elongation factor Tu. AB - The RNA genome of coliphage Qbeta is replicated by a complex of four proteins, one of them being the translation elongation factor Tu. The role of EF-Tu in this RNA polymerase complex is still unclear, but the obligate presence of translationally functional EF-Tu in the cell hampers the use of conventional mutational analysis. Therefore, we designed a system based on affinity chromatography and could separate two types of complexes by placing an affinity tag on mutated EF-Tu species. Thus, we were able to show a direct link between the vital tRNA binding property of EF-Tu and polymerase activity. PMID- 12651025 TI - Expression and purification of human cardiac troponin subunits and their functional incorporation into isolated cardiac mouse myofibrils. AB - The three subunits of the human cardiac troponin complex (hcTnC, hcTnI, hcTnT) were overexpressed in E. coli, purified and reconstituted to form the hcTn complex. This complex was then incorporated into subcellular bundles of mouse cardiac myofibrils whereby the native mcTn complex was replaced. On thus exchanged myofibrils, isometric force kinetics following sudden changes in free Ca(2+) concentration were measured using atomic force cantilevers. Following the exchange, the myofibrillar force remained fully Ca(2+) regulated, i.e. myofibrils were completely relaxed at pCa 7.5 and developed the same maximum Ca(2+) activated isometric force upon increasing the pCa to 4.5 as unexchanged myofibrils. The replacement of endogenous mcTn by wild-type hcTn neither altered the kinetics of Ca(2+)-induced force development of the mouse myofibrils nor the kinetics of force relaxation induced by the sudden, complete removal of Ca(2+). Preparations of functional Tn reconstituted myofibrils provide a promising model to study the role of Tn in kinetic mechanisms of cardiac myofibrillar contraction and relaxation. PMID- 12651026 TI - Production of a soluble gammadelta T-cell receptor to identify ligands for the murine intestinal intraepithelial gammadelta T cell population. AB - Although the functions and antigen recognition requirements of alphabeta T cells are well characterised, the antigens recognised by gammadelta T cells and the consequences of this recognition are unclear. gammadelta T cells are enriched within epithelia, where they eradicate transformed epithelial cells and regulate inflammation. To understand how this occurs, we need to understand the cellular ligands recognised by the gammadelta cell through the gammadelta T-cell receptor (TCR). We have therefore generated a soluble TCR (sTCR) to identify ligands for the murine gammadelta intestinal intraepithelial lymphocyte (IEL) population. sTCR was produced in the baculovirus expression system and purified by affinity chromatography on an anti-TCRdelta affinity column. sTCR was recognised by a panel of conformation-specific anti-TCRgammadelta antibodies. We will now use our sTCR to directly test the binding of putative ligands to the TCR using surface plasmon resonance, and to isolate the ligand biochemically. PMID- 12651027 TI - On-column refolding of an insoluble histidine tag recombinant exopolyphosphatase from Trypanosoma brucei overexpressed in Escherichia coli. AB - An exopolyphosphatase gene has been cloned by polymerase chain reaction (PCR) from Trypanosoma brucei and the corresponding protein overexpressed as a recombinant His-tag (histidine tag) exopolyphosphatase in E. coli in order to characterize its biochemical activity and to produce antibody to determine its localization. Because overexpression of this protein in bacteria resulted in the formation of inactive inclusion bodies, these structures were first solubilized in denaturant condition (6 M urea). Secondly, after a capture step using immobilized metal affinity chromatography (IMAC), a gradual refolding of the protein was performed on-column from 6 M to 0 M urea in the presence of 1% Triton X-100. Triton X-100 was used to abolish protein aggregation during the refolding step. The purified enzyme was active, demonstrating that at least part of the proteins was properly refolded. PMID- 12651028 TI - Identification of a tubulin binding motif on the P2X2 receptor. AB - To isolate proteins interacting with P2X receptors, GST fusion proteins containing the intracellular C terminal tail of P2X(2), P2X(5), or P2X(7) were used as bait to screen detergent extracts of rat brain synaptosomes. By SDS-PAGE combined with mass spectrometry, two interacting proteins were identified: betaIII tubulin and myelin basic protein. While myelin basic protein bound to all three P2X subunits, betaIII tubulin interacted exclusively with the P2X(2) subunit. The tubulin binding domain could be confined to a proline-rich segment (amino acids 371-412) of the P2X(2) subunit. Our results suggest a role for microtubules in the cellular localisation of the P2X(2) receptor. PMID- 12651029 TI - GST-Fbe can recognize beta-chains of fibrin(ogen) on explanted materials. AB - Staphylococcus epidermidis, a coagulase-negative staphylococcus (CoNS), is one of the leading pathogens of nosocomial infections, particularly associated with foreign body infections. Adherence of S. epidermidis to fibrinogen deposited on the surfaces of implants is important for the development of foreign body infections. A gene (fbe) encoding a fibrinogen-binding protein from S. epidermidis (Fbe) was identified by shotgun phage display. A portion of fbe was cloned into a GST-fusion vector. Affinity to glutathione-Sepharose by the GST-tag and affinity to fibrinogen-Sepharose by the Fbe part were applied to purify the recombinant Fbe. The purity and efficacy of the methods used in protein purification was compared. Furthermore, the potential physiological role of Fbe was studied by the interaction between GST-Fbe and components extracted from explanted materials in vitro. PMID- 12651030 TI - Production, purification and characterisation of genetically derived scFv and bifunctional antibody fragments capable of detecting illicit drug residues. AB - We have generated a single chain antigen binding protein (scFv) recognising morphine. Variable regions of heavy (V(H)) and light (V(L)) chain antibody genes isolated from a murine immune repertoire were connected via a glycine-serine linker and cloned into the expression vector pAK 400. The scFv was produced in Escherichia coli JM83 yielding a functional protein of approximately M(r) 30000. Immunoaffinity chromatography using M3G-BSA-Sepharose column proved most effective for scFv purification. Purity was monitored by SDS-PAGE and Western blotting and the scFv characterised using ELISA and BIAcore. The scFv was capable of specifically binding free morphine in solution and was applicable to real sample analysis in saliva. In order to express a bivalent "minibody" the scFv gene was recloned into a vector containing a gene encoding a helix for dimerisation. The scFv was expressed as a protein of M(r) 75000 and retained its antibody binding capabilities. Cloning the scFv gene into a vector containing the bacterial alkaline phosphatase gene produced a bifunctional molecule, which retained the binding activity of the parental scFv along with the enzymatic activity of alkaline phosphatase. PMID- 12651031 TI - Semi automated production of a set of different recombinant GST-Streptag fusion proteins. AB - We describe a high-throughput procedure for the large-scale production of recombinant GST-Streptag fusion proteins. This three-step process, comprising cloning, expression and purification, simultaneously produces up to 96 different proteins in a multi-well format with high yield and purity. Two complementary oligonucleotides, together encoding a specific peptide sequence are annealed and directly ligated into a pre-digested pGEX-2T plasmid carrying an N-terminal GST tag and a C-terminal Streptag. Following expression, a multichannel pipetting robot purifies the resulting fusion proteins within 2 h by affinity chromatography on Streptactin Macroprep mini-columns. PMID- 12651032 TI - Choice of prosthetic heart valve for adult patients. AB - This review summarizes the major long-term (> or =10 to 15 years) patient outcomes after insertion of many Food and Drug Administration approved prosthetic heart valves (PHV). Mechanical PHV was associated with a better survival (p < 0.02) at 15 years after aortic valve replacement (AVR) than with a bioprosthesis in the Department of Veterans Affairs (DVA) trial. In both the DVA and the Edinburgh Heart Valve trials, bioprosthesis were associated with structural valve deterioration (SVD) (mitral valve replacement [MVR] > AVR) and, therefore, for replacement of the PHV. Thromboembolism and bleeding rate were higher with mechanical PHV. Mortality after AVR and MVR is high at 10 to 15 years because of the associated comorbid conditions and older age of patients. Outcomes with "new" good valves are similar to that with "older" good valves. Complication rates of thromboembolism, bleeding, endocarditis, and leak vary widely; the rates of these complications are not different among different mechanical PHV and among different bioprosthetic PHV. Structural valve deterioration is rare with mechanical PHV. Structural valve deterioration of bioprosthesis after MVR is higher than after AVR; after AVR, homografts and bioprosthesis have similar rates of SVD. The exact rate of SVD of the pulmonary autograft is uncertain. Valve prosthesis-patient mismatch is clinically important when it is severe and in selected patients when it is moderate. Bioprosthesis have a low rate of SVD in the older patient and, thus, are the PHV of choice for AVR in patients > or =60 to 65 years of age and for MVR in patients > or =65 to 70 years of age; in younger patients mechanical valves are the PHV of choice. In individual patients there may be exceptions to these general rules. PMID- 12651033 TI - Quantitative analysis of the admission electrocardiogram identifies patients with unstable coronary artery disease who benefit the most from early invasive treatment. AB - OBJECTIVES: The aim of the present study was to evaluate whether the effect of an early invasive treatment strategy differed between patients sub-grouped according to their severity of myocardial ischemia, as evaluated by quantitative electrocardiographic (ECG) analysis at the time of presentation. The present study is a sub-study of the previously published Fast Revascularization during InStability in Coronary artery disease trial (FRISC-II). BACKGROUND: An early invasive treatment strategy has been shown to be the preferable treatment for non ST-segment elevation acute coronary syndromes (ACS). The population of patients with unstable coronary artery disease is heterogeneous regarding both the underlying pathology and prognosis. Early risk stratification is important to select patient subgroups that will benefit the most from a given treatment. METHODS: In 2,201 patients with non-ST-segment elevation ACS, the ischemic burden at hospital admission was determined by quantitative measurements of ST-T-segment deviations on the ECG. The patients were subsequently sub-grouped in tertiles based on the amount of ST-segment deviation. The primary end point for this analysis was death or myocardial infarction (MI) within one year after study inclusion. RESULTS: The invasive treatment strategy produced a reduction of approximately 50% in death or MI among the patients with intermediate or major ST segment deviation. The findings were independent of age, gender, or troponin T status. The patients with confounding factors precluding ST analysis had a poor outcome regardless of the treatment strategy. CONCLUSIONS: Ischemic burden on the admission ECG identifies patients with ACS who benefit the most from an invasive treatment strategy. When the standard ECG is scrutinized with complete quantitative measurements, it provides independent information on prognosis and benefit of treatment. PMID- 12651034 TI - Troponin and C-reactive protein have different relations to subsequent mortality and myocardial infarction after acute coronary syndrome: a GUSTO-IV substudy. AB - OBJECTIVES: We sought to evaluate C-reactive protein (CRP) and troponin T (TnT) as predictors of risk of the individual end points of mortality and myocardial infarction (MI) in a large cohort of patients with acute coronary syndrome (ACS). BACKGROUND: Both CRP and TnT predict risk of future coronary events in patients with ACS. However, the relationships between the levels of the markers and the individual end points are still unclear. METHODS: Baseline levels of CRP and TnT were determined in 7,108 patients with ACS not undergoing early revascularization in the Global Use of Strategies To Open occluded arteries trial IV (GUSTO-IV) trial and related to outcome at 30 days. RESULTS: Quartiles of TnT related to 30 day mortality, which was 1.1%, 3.7%, 3.7%, and 7.4% (p < 0.001) and to the rate of MI: 2.5%, 6.7%, 7.2%, and 5.6% (p < 0.001). Quartiles of CRP also related to 30-day mortality, which was 2.0%, 3.3%, 3.9%, and 6.3% (p < 0.001), whereas there was no relationship to the 30-day rate of MI: 5.6%, 4.7%, 5.2%, and 5.9% (p = 0.48). On multivariable analysis, both TnT and CRP were independent predictors of mortality, but only TnT was a predictor of MI. The combination of CRP and TnT provides an even better risk stratification of mortality, with 0.3% and 9.1% death rates, respectively, when both markers are in the bottom versus top quartiles. CONCLUSIONS: In ACS, baseline levels of TnT and CRP are independently related to 30-day mortality. Any detectable elevation of TnT, but not of CRP, is also associated with an increased risk of subsequent MI. Regarding mortality, the combination of both markers provides a better risk stratification than either one alone. PMID- 12651035 TI - Thrombolysis in myocardial infarction myocardial perfusion grade in angiography correlates with myocardial salvage in patients with acute myocardial infarction treated with stenting or thrombolysis. AB - OBJECTIVES: We sought to assess the relationship between the Thrombolysis In Myocardial Infarction (TIMI) myocardial perfusion (TMP) grade and myocardial salvage as well as the usefulness of TMP grade in comparing two different reperfusion strategies. BACKGROUND: The angiographic index of TMP grade correlates with infarct size and mortality after thrombolysis for acute myocardial infarction (AMI). Its relationship to myocardial salvage and its usefulness in comparing different reperfusion strategies are not known. METHODS: We analyzed the TMP grade on angiograms obtained at one to two weeks after treatment in 267 patients enrolled in two randomized trials that compared stenting with thrombolysis in AMI. Patients were classified into two groups: 159 patients with TMP grade 2/3 and 108 patients with TMP grade 0/1. Two scintigraphic studies were performed: before and one to two weeks after reperfusion. The salvage index was calculated as the proportion of the area at risk salvaged by reperfusion. RESULTS: Patients with TMP grade 2/3 had a higher salvage index (0.49 +/- 0.42 vs. 0.34 +/- 0.49, p = 0.01), a smaller final infarct size (15.4 +/- 15.5% vs. 22.1 +/- 16.2% of the left ventricle, p = 0.001), and a trend toward lower one-year mortality (3.8% vs. 8.3%, p = 0.11) than patients with TMP grade 0/1. The relationship between TMP and salvage index was independent of the form of reperfusion therapy. The proportion of patients with TMP grade 2/3 was significantly higher after stenting than after thrombolysis (70.9% vs. 48.1%, p = 0.001). CONCLUSIONS: These findings show that the TMP grade is a useful marker of the degree of myocardial salvage achieved with reperfusion and a sensitive indicator of the efficacy of reperfusion strategies in patients with AMI. PMID- 12651036 TI - The T-786C endothelial nitric oxide synthase genotype is a novel risk factor for coronary artery disease in Caucasian patients of the GENICA study. AB - OBJECTIVES: We investigated the association of polymorphisms in the promoter region and exon 7 endothelial nitric oxide synthase (eNOS) gene with coronary artery disease (CAD). BACKGROUND: Endothelial dysfunction foretells cardiovascular events and can be genetically determined. METHODS: We genotyped for the promoter (T(-786)C) and exon 7 (Glu298Asp, G(894)T) polymorphisms in 1,225 subjects; 1,106 were consecutive patients undergoing coronary angiography and 119 control subjects without any cardiovascular risk factors. Genotyping was performed with melting curve analysis of polymerase chain reaction products from allele-specific acceptor and donor probes that were 5'- and 3'-end labeled with LCRed640 and fluorescein, respectively; CAD was assessed by quantitative coronary angiography. We performed multiple logistic regression analysis for the effect of the T(-786)C, the missense Glu298Asp variant, and other coronary risk factors on two- and three-vessel CAD. RESULTS: The overall genotype distribution of T(-786)C (CC = 17.7%, CT = 40.4%, and TT = 41.9%) and Glu298Asp (GG = 43.3%, GT = 37.0%, and TT = 19.7%) was consistent with the Hardy-Weinberg equilibrium. The regression analysis showed that the T(-786)C, but not the missense Glu298Asp variant, significantly predicted CAD, independent of other risk factors. Compared with TT homozygous, subjects carrying the C allele had a significant (p = 0.002) increase in the odds ratio of harboring two- or three-vessel CAD of 1.672 (95% confidence interval, 1.062 to 2.527). A subgroup analysis confirmed this effect of the T(-786)C polymorphism in men (p = 0.007), cigarette smokers (p = 0.001), subjects older than 60 years of age (p = 0.007), with hypercholesterolemia (p = 0.011), low high-density lipoprotein cholesterol (p = 0.006), and overweight or with obesity (p = 0.041). CONCLUSIONS: The C allele at the T(-786)C endothelial nitric oxide synthase polymorphism is associated with a higher risk of multivessel CAD in Caucasians. PMID- 12651038 TI - Functional polymorphisms in a candidate gene for atherothrombosis: unraveling the complex fabric of a polygenic phenotype. PMID- 12651037 TI - The T-786C and Glu298Asp polymorphisms of the endothelial nitric oxide gene affect the forearm blood flow responses of Caucasian hypertensive patients. AB - OBJECTIVES: We sought to investigate whether two polymorphisms located in the promoter (T(-786)C) and exon 7 (Glu298Asp) of the endothelial nitric oxide (NO) synthase (eNOS) gene affected agonists-mediated NO release. BACKGROUND: Endothelial dysfunction can be genetically determined. Therefore, we investigated whether two polymorphisms located in the eNOS gene affected agonists-mediated NO release. METHODS: We compared endothelial-dependent and -independent vasodilation of the different eNOS genotypes in a cross-sectional study on 187 subjects, of whom 137 were uncomplicated essential hypertensive patients (PH) (49 +/- 9 years, 151 +/- 11/99 +/- 5 mm Hg) and 50 healthy normotensive subjects (NT) (43 +/- 16 years, 123 +/- 10/78 +/- 7 mm Hg). Endothelial-dependent and -independent vasodilation was assessed as the forearm blood flow response to incrementally increasing doses of acetylcholine (0.15, 0.45, 1.5, 4.5, 15 microg/100 ml/min) and sodium nitroprusside (1, 2, 4 microg/100 ml/min), respectively. Genotyping was performed with melting curve analysis (Lightcycler) of polymerase chain reaction products from acceptor (5' end-labeled with LCRed 640) and donor probes (3' end-labeled with fluorescein) specific for each polymorphism. The genotype distribution of T(-786)C (CC = 21.9%, CT = 48.7%, TT = 29.4%) and Glu298Asp (GG = 39.0%, GT =51.9%, TT = 9.1%) was similar in PH and NT. A repeated measure analysis of variance showed a blunting of endothelium-dependent vasodilation in PH compared with NT (p < 0.001). A significant effect of the T(-786)C (p = 0.002) but not of the Glu298Asp (p = NS) eNOS polymorphism on endothelial-dependent vasodilation was found. However, we also detected a significant interaction between the T(-786)C and Glu298Asp polymorphism (p < 0.001). No effect on either polymorphism on endothelial-independent vasodilation was seen. CONCLUSIONS: The T(-786)C promoter polymorphism and its interaction with exon 7 Glu298Asp affect endothelium-dependent vasodilation in mild-to-moderate PH patients and NT Caucasian subjects. PMID- 12651039 TI - The influence of left ventricular hypertrophy on survival in patients with coronary artery disease: do race and gender matter? AB - OBJECTIVES: We sought to determine the overall prognostic importance of left ventricular hypertrophy (LVH) among patients with coronary artery disease (CAD), as well as to determine whether this risk varies as a function of race or gender. BACKGROUND: Left ventricular hypertrophy is more prevalent among blacks and women than their counterparts. Blacks and women also have higher mortality with coronary disease. METHODS: We studied records of 2,461 patients (19% black, 42% women) diagnosed with CAD at cardiac catheterization between 1990 and 1998 from a single academic center. Left ventricular hypertrophy was defined using standard echocardiographic measures. Cox proportional hazards models were used for adjusted survival analyses. Mean patient follow-up was three years. RESULTS: Patients with LVH were older (68 vs. 65 years, p < 0.01), more often women (54% vs. 36%, p < 0.01), and black (25% vs. 16%, p < 0.01), and had higher unadjusted three-year mortality rates than patients without LVH (42% vs. 34%, p < 0.01). Left ventricular hypertrophy remained an independent predictor of mortality after adjusting for other clinical risk factors (hazard ratio 1.56, 95% confidence interval 1.35 to 1.80) with prognostic importance equivalent to that of left ventricular ejection fraction. Although the relative risk of LVH did not vary by race or gender, the attributable risk of LVH was greater in blacks and women. CONCLUSIONS: Clinicians should consider the prognostic importance of LVH when assessing risk in patients with CAD. Because LVH is more common among black and women patients with CAD, it partially accounts for racial and gender differences in survival. PMID- 12651040 TI - Influence of systolic blood pressure and body mass index on left ventricular structure in healthy African-American and white young adults: the CARDIA study. AB - OBJECTIVES: In the Coronary Artery Risk Development in Young Adults (CARDIA) Study comprised of a generally healthy, biracial cohort of 28- to 40-year-old adults, we sought to characterize the distribution of left ventricular (LV) mass and LV geometry and the relationship of systolic blood pressure (SBP), body mass index (BMI), and fasting insulin to LV mass and geometry. BACKGROUND: Left ventricular mass is a risk factor for cardiovascular morbidity and mortality. METHODS: Two-dimensionally guided M-mode echocardiograms were used to calculate LV mass index (g/height(2.7)) and geometry. RESULTS: Black men had highest LV mass index followed by white men, black women, and white women. Blacks had higher LV wall thickness/diameter ratios than whites. Left ventricular hypertrophy was present in 2% of the cohort. Going from highest to lowest quartile for LV mass index and LV wall thickness/diameter ratio, SBP and BMI were highest in those with the highest LV mass index and LV wall thickness/diameter ratio. Increasing BMI and SBP over a 10-year interval was also strongly related to LV structure in most race/gender groups. CONCLUSIONS: In a generally healthy young adult cohort, LV structure as defined by LV mass and geometry is associated with SBP and BMI at levels generally considered normal. PMID- 12651041 TI - A prospective, blinded determination of the natural history of aspirin resistance among stable patients with cardiovascular disease. AB - OBJECTIVES: This study was designed to determine if aspirin resistance is associated with clinical events. BACKGROUND: Aspirin resistance, defined by platelet function testing and presumed clinical unresponsiveness to aspirin, has been previously reported by our group and others. However, little information exists linking the laboratory documentation of aspirin resistance and long-term clinical events. METHODS: We prospectively enrolled 326 stable cardiovascular patients from 1997 to 1999 on aspirin (325 mg/day for > or =7 days) and no other antiplatelet agents. We tested for aspirin sensitivity by optical platelet aggregation using adenosine diphosphate (ADP) and arachidonic acid (AA). The primary outcome was the composite of death, myocardial infarction (MI), or cerebrovascular accident (CVA). Mean follow-up was 679 +/- 185 days. Aspirin resistance was defined as a mean aggregation of > or =70% with 10 microM ADP and > or =20% with 0.5 mg/ml AA. RESULTS: Of the patients studied, 17 (5.2%) were aspirin resistant and 309 (94.8%) were not aspirin resistant. During follow-up, aspirin resistance was associated with an increased risk of death, MI, or CVA compared with patients who were aspirin sensitive (24% vs. 10%, hazard ratio [HR] 3.12, 95% confidence interval [CI] 1.10 to 8.90, p = 0.03). Stratified multivariate analyses identified platelet count, age, heart failure, and aspirin resistance to be independently associated with major adverse long-term outcomes (HR for aspirin resistance 4.14, 95% CI 1.42 to 12.06, p = 0.009). CONCLUSIONS: This study demonstrates the natural history of aspirin resistance in a stable population, documenting a greater than threefold increase in the risk of major adverse events associated with aspirin resistance. PMID- 12651042 TI - Aspirin resistance: a new independent predictor of vascular events? PMID- 12651043 TI - A randomized comparison of clopidogrel and aspirin versus ticlopidine and aspirin after the placement of coronary artery stents. AB - OBJECTIVES: The aim of the present study was to compare clopidogrel and ticlopidine after coronary stenting with regard to cardiovascular death during long-term follow-up. BACKGROUND: Randomized trials comparing clopidogrel and ticlopidine with a restricted use of intravenous glycoprotein IIb/IIIa inhibition have reported a trend toward a higher incidence of thrombotic stent occlusion with clopidogrel at 30 days. METHODS: After successful coronary stent implantation, 700 patients with 899 lesions were randomly assigned to receive a four-week course of either 500 mg ticlopidine (n = 345) or 75 mg clopidogrel (n = 355) in addition to 100 mg aspirin. Cardiovascular death was the primary end point and was recorded during a median follow-up period of 28 months. RESULTS: Cardiovascular death occurred in eight patients with ticlopidine versus 26 patients with clopidogrel (hazard ratio with ticlopidine compared with clopidogrel, 0.30; 95% confidence interval [CI], 0.14 to 0.66; p = 0.003). After adjustment for co-variables, ticlopidine reduced the risk of cardiovascular death by 63% compared with clopidogrel. The combined end point of cardiovascular death or nonfatal myocardial infarction was present in 19 patients assigned ticlopidine, compared with 40 patients assigned clopidogrel (hazard ratio, 0.45; p = 0.005). The hazard ratio for all-cause mortality with ticlopidine as compared with clopidogrel was 0.30 (95% CI, 0.14 to 0.64; p = 0.002). CONCLUSIONS: After the placement of coronary artery stents in unselected patients, ticlopidine was associated with a significantly lower mortality than clopidogrel. This raises concern about the current practice of substituting clopidogrel for ticlopidine after stenting and highlights the need for further long-term studies. PMID- 12651044 TI - Relationship of race to sudden cardiac death in competitive athletes with hypertrophic cardiomyopathy. AB - OBJECTIVES: The goal of this study was to determine the impact of race on identification of hypertrophic cardiomyopathy (HCM). BACKGROUND: Sudden death in young competitive athletes is due to a variety of cardiovascular diseases (CVDs) and, most commonly, HCM. These catastrophes have become an important issue for African Americans, although HCM has been previously regarded as rare in this segment of the U.S. population. METHODS: We studied the relationship of race to the prevalence of CVDs causing sudden death in our national athlete registry, and compared these findings with a representative multicenter hospital-based cohort of patients with HCM. RESULTS: Of 584 athlete deaths, 286 were documented to be due to CVD at ages 17 +/- 3 years; 156 (55%) were white, and 120 (42%) were African American. Most were male (90%), and 67% participated in basketball and football. Among the 286 cardiovascular deaths, most were due to HCM (n = 102; 36%) or anomalous coronary artery of wrong sinus origin (n = 37; 13%). Of the athletes who died of HCM, 42 (41%) were white, but 56 (55%) were African American. In contrast, of 1,986 clinically identified HCM patients, only 158 (8%) were African American (p < 0.001). CONCLUSIONS: In this autopsy series, HCM represented a common cause of sudden death in young and previously undiagnosed African American male athletes, in sharp contrast with the infrequent clinical identification of HCM in a hospital-based population (i.e., by seven-fold). This discrepancy suggests that many HCM cases go unrecognized in the African American community, underscoring the need for enhanced clinical recognition of HCM to create the opportunity for preventive measures to be employed in high-risk patients with this complex disease. PMID- 12651045 TI - Variable clinical manifestation of a novel missense mutation in the alpha tropomyosin (TPM1) gene in familial hypertrophic cardiomyopathy. AB - OBJECTIVES: This study was initiated to identify the disease-causing genetic defect in a family with hypertrophic cardiomyopathy (HCM) and high incidence of sudden death. BACKGROUND: Familial hypertropic cardiomyopathy (FHC) is an autosomal dominant transmitted disorder that is genetically and clinically heterogeneous. Mutations in 11 genes have been associated with the pathogenesis of the disease. METHODS: We studied a large FHC family, first by linkage analysis, to identify the gene involved, and subsequently screened the gene, encoding alpha-tropomyosin (TPM1), for mutations by using single-strand conformation polymorphism and sequencing analysis. RESULTS: Twelve family members presented clinical features of HCM, five of whom died at young age, while others had only mild clinical features. Marker analysis showed linkage for the TPM1 gene on chromosome 15q22 (maximal logarithm of the odds score is 5.16, theta = 0); subsequently, a novel missense mutation (Glu62Gln) was identified. CONCLUSIONS: The novel mutation identified in TPM1 is associated with the clinical features of cardiac hypertrophy in all but one genetically affected member of this large family. The clinical data suggest a malignant phenotype at young age with a variable clinical manifestation and penetrance at older age. The Glu62Gln mutation is the sixth TPM1 mutation identified as the cause of FHC, indicating that mutations in this gene are very rare. This is the first reported amino acid substitution at the f-position within the coiled-coil structure of the tropomyosin protein. PMID- 12651046 TI - Hypertrophic cardiomyopathy in a large community-based population: clinical outcome and identification of risk factors for sudden cardiac death and clinical deterioration. AB - OBJECTIVES: This study evaluates the clinical course and identifies risk factors for sudden cardiac death (SCD) and clinical deterioration in hypertrophic cardiomyopathy (HCM) in a large community-based population. Comparison was made with data from six tertiary referral and six nonreferral institutions. BACKGROUND: Hypertrophic cardiomyopathy is a disease with marked heterogeneity in clinical presentation and prognosis. Risk factors for SCD are not well defined in patients free of referral bias. METHODS: Between 1970 and 1999, 225 consecutive patients (mean age [+/-SD] 41+/-16 years) were examined and followed at yearly intervals. RESULTS: Forty-four deaths were recorded of which 27 cases were cardiovascular. Fourteen patients died suddenly, six were successfully resuscitated, and seven patients died of congestive heart failure. The annual mortality, annual cardiac mortality, and annual mortality due to sudden death were 1.3%, 0.8%, and 0.6%, respectively. At least one New York Heart Association (NYHA) functional class deterioration was reported in 33% of the patients with a significant (> or =50 mm Hg) left ventricular outflow tract (LVOT) gradient in contrast to 7% without obstruction. The presence of syncope was related to SCD (p < 0.05). Younger age and more severe functional limitation distinguishes patients from in hospital-based centers from the ones in community-based centers. CONCLUSIONS: Hypertrophic cardiomyopathy is a benign disease in an unselected population with a low incidence of cardiac death. Syncope was associated with a higher incidence of SCD and patients with a significant LVOT obstruction were more susceptible to clinical deterioration. PMID- 12651047 TI - On predictors of sudden cardiac death in hypertrophic cardiomyopathy. PMID- 12651048 TI - Heart failure etiology and response to milrinone in decompensated heart failure: results from the OPTIME-CHF study. AB - OBJECTIVES: The goal of this study was to assess the interaction between heart failure (HF) etiology and response to milrinone in decompensated HF. BACKGROUND: Etiology has prognostic and therapeutic implications in HF, but its relationship to response to inotropic therapy is unknown. METHODS: The Outcomes of a Prospective Trial of Intravenous Milrinone for Exacerbations of Chronic Heart Failure (OPTIME-CHF) study randomized 949 patients with systolic dysfunction and decompensated HF to receive 48 to 72 h of intravenous milrinone or placebo. The primary end point was days hospitalized from cardiovascular causes within 60 days. In a post-hoc analysis, we evaluated the interaction between response to milrinone and etiology of HF. RESULTS: The primary end point was 13.0 days for ischemic patients and 11.7 days for nonischemic patients (p = 0.2). Sixty-day mortality was 11.6% for the ischemic group and 7.5% for the nonischemic group (p = 0.03). After adjustment for baseline differences, there was a significant interaction between etiology and the effect of milrinone. Milrinone-treated patients with ischemic etiology tended to have worse outcomes than those treated with placebo in terms of the primary end point (13.6 days for milrinone vs. 12.4 days for placebo, p = 0.055 for interaction) and the composite of death or rehospitalization (42% vs. 36% for placebo, p = 0.01 for interaction). In contrast, outcomes in nonischemic patients treated with milrinone tended to be improved in terms of the primary end point (10.9 vs. 12.6 days placebo) and the composite of death or rehospitalization (28% vs. 35% placebo). CONCLUSIONS: Milrinone may have a bidirectional effect based on etiology in decompensated HF. Milrinone may be deleterious in ischemic HF, but neutral to beneficial in nonischemic cardiomyopathy. PMID- 12651049 TI - Age and hemodynamic responses to tilt testing in those with syncope of unknown origin. AB - OBJECTIVES: We sought to determine whether there are age-related differences in vasovagal syncope. BACKGROUND: In those with suspected vasovagal (neurocardiogenic) syncope, tilt testing demonstrates different hemodynamic responses. These responses may be age related, reflecting differing underlying pathophysiology. METHODS: Using a two-stage tilt protocol with glyceryl trinitrate (GTN) provocation, 505 consecutive syncopal patients were studied. Their baseline characteristics and hemodynamic responses during both early and tilt-induced collapse were analyzed. Hemodynamic responses were classified using the VAsovagal Syncope International Study (VASIS) criteria: mixed, cardioinhibition, severe cardioinhibition/asystole, pure vasodepression, chronotropic incompetence, and excessive heart rate rise. Multivariate regression analyses were performed to determine the associations of the baseline clinical characteristics (including age) and the tilt-induced hemodynamic responses. RESULTS: Thirty-three patients were unable to tolerate tilt testing. Age was independently associated with distinct responses during tilt. Chronotropic incompetence was predicted by increasing age (odds ratio [OR] 1.04, p < 0.0002). Younger age predicted an excessive heart rate rise (OR 0.97, p < 0.0005). Pure vasodepression was more common in the older group (>65 years; OR 29.5, p < 0.0001), whereas severe cardioinhibition was much less common in the older age group (OR 0.18, p < 0.0001). CONCLUSIONS: There appear to be distinct pathophysiologies underlying vasovagal syncope in different age groups. Young people appear to have excessive cardiac and autonomic responses to stress, whereas older patients appear to have a more generalized cardiovascular decline, with attenuated cardiac and autonomic responses to stress. PMID- 12651050 TI - The prevalence of coronary artery calcium among diabetic individuals without known coronary artery disease. AB - OBJECTIVES: We sought to examine the age and gender distribution of coronary artery calcium (CAC) by diabetes status in a large cohort of asymptomatic individuals. BACKGROUND: Among individuals with diabetes, coronary artery disease (CAD) is a major cause of morbidity and mortality. Electron-beam tomography (EBT) quantifies CAC, a marker for atherosclerosis. METHODS: Screening for CAC by EBT was performed in 30,904 asymptomatic individuals stratified by their self reported diabetes status, gender, and age. The distribution of CAC across the strata and the association between diabetes and CAC were examined. RESULTS: Compared with nondiabetic individuals (n = 29,829), those with diabetes (n = 1,075) had higher median CAC scores across all but two age groups (women 40 to 44 years old and men and women > or =70 years old). Overall, the likelihood of having a CAC score in the highest age/gender quartile was 70% greater for diabetic individuals than for their nondiabetic counterparts. CONCLUSIONS: Younger diabetic individuals appear to have calcified plaque burden comparable to that of older individuals without diabetes. These findings call for future research to determine if EBT-CAC screening has an incremental value over the current CAD risk assessment of individuals with diabetes. PMID- 12651051 TI - Acute, short-term hyperglycemia enhances shear stress-induced platelet activation in patients with type II diabetes mellitus. AB - OBJECTIVES: The aim of our study was to assess whether acute, short-term hyperglycemia affects platelet reactivity in patients with Type II diabetes mellitus (T2DM). BACKGROUND: Hyperglycemic spikes are thought to precipitate ischemic events in T2DM. Previous studies have shown in vivo platelet activation in diabetes; however, no studies have assessed whether acute in vivo hyperglycemia induces further activation of platelets. METHODS: In a cross-over, randomized, double-blind study, 12 patients with T2DM underwent 4 h of either acute hyperglycemia (13.9 mmol/l, 250 mg/dl) or euglycemia (5.5 mmol/l, 100 mg/dl). Shear stress-induced platelet activation, P-selectin and lysosomal integral membrane protein (LIMP) expression on platelets in the bleeding-time blood, urinary 11-dehydro-thromboxane B(2) (TxB(2)) excretion, von Willebrand factor:antigen (vWF:Ag), and von Willebrand factor:activity (vWF:activity) were measured before and after hyperglycemia or euglycemia. RESULTS: Shear stress induced platelet activation, P-selectin and LIMP expression on platelets in the bleeding-time blood, and urinary 11-dehydro-TxB(2) excretion increased significantly after hyperglycemic clamping, whereas no changes were observed after euglycemic clamping. Plasma vWF:Ag and vWF:activity increased strikingly in parallel fashion after hyperglycemic clamping, whereas no changes were observed after euglycemic clamping. CONCLUSIONS: Our data demonstrate that acute, short term hyperglycemia induces an increased activation of platelets exposed to high shear stress conditions in vitro (filtration method) or in vivo (bleeding time). In vivo platelet activation is reflected by an increased urinary excretion of 11 dehydro-TxB(2). The increased levels of vWF in the circulation correlate with the increase in platelet activation markers and may indicate some degree of causation. Acute, short-term hyperglycemia in T2DM may precipitate vascular occlusions by facilitating platelet activation. PMID- 12651052 TI - A simple method for noninvasive estimation of pulmonary vascular resistance. AB - OBJECTIVES: We sought to test whether the ratio of peak tricuspid regurgitant velocity (TRV, ms) to the right ventricular outflow tract time-velocity integral (TVI(RVOT), cm) obtained by Doppler echocardiography (TRV/TVI(RVOT)) provides a clinically reliable method to determine pulmonary vascular resistance (PVR). BACKGROUND: Pulmonary vascular resistance is an important hemodynamic variable used in the management of patients with cardiovascular and pulmonary disease. Right-heart catheterization, with its associated disadvantages, is required to determine PVR. However, a reliable noninvasive method is unavailable. METHODS: Simultaneous Doppler echocardiographic examination and right-heart catheterization were performed in 44 patients. The ratio of TRV/TVI(RVOT) was then correlated with invasive PVR measurements using regression analysis. An equation was modeled to calculate PVR in Wood units (WU) using echocardiography, and the results were compared with invasive PVR measurements using the Bland Altman analysis. Using receiver-operating characteristics curve analysis, a cutoff value for the Doppler equation was generated to determine PVR >2WU. RESULTS: As calculated by Doppler echocardiography, TRV/TVI(RVOT) correlated well (r = 0.929, 95% confidence interval 0.87 to 0.96) with invasive PVR measurements. The Bland-Altman analysis between PVR obtained invasively and that by echocardiography, using the equation: PVR = TRV/TVI(RVOT) x 10 + 0.16, showed satisfactory limits of agreement (mean 0 +/- 0.41). A TRV/TVI(RVOT) cutoff value of 0.175 had a sensitivity of 77% and a specificity of 81% to determine PVR >2WU. CONCLUSIONS: Doppler echocardiography may provide a reliable, noninvasive method to determine PVR. PMID- 12651053 TI - Pulmonary function in primary pulmonary hypertension. AB - OBJECTIVES: The study was done to ascertain the degree to which abnormalities in resting lung function correlate with the disease severity of patients with primary pulmonary hypertension (PPH). BACKGROUND: Patients with PPH are often difficult to diagnose until several years after the onset of symptoms. Despite the seriousness of the disorder, the diagnosis of PPH is often delayed because it is unsuspected and requires invasive measurements. Although PPH often causes abnormalities in resting lung function, these abnormalities have not been shown to be statistically significant when correlated with other measures of PPH severity. METHODS: Resting lung mechanics and diffusing capacity for carbon monoxide DL(CO) were assessed in 79 patients whose findings conformed to the classical diagnostic criteria of PPH and who had no evidence of secondary causes of pulmonary hypertension. These findings were correlated with severity of disease as assessed by cardiac catheterization, New York Heart Association (NYHA) class, and cardiopulmonary exercise testing. RESULTS: When PPH patients were first evaluated at our referral clinic, the DL(CO) and lung volumes were decreased in approximately three-quarters and one-half, respectively. The decreases in DL(CO), and to a lesser extent lung volumes, correlated significantly with decreases in peak oxygen uptake (reflecting maximum cardiac output), peak oxygen pulse (reflecting maximum stroke volume), and anaerobic threshold (reflecting sustainable exercise capacity) and higher NYHA class. CONCLUSIONS: Patients with PPH commonly have abnormalities in lung mechanics and DL(CO) levels that correlate significantly with disease severity. These measurements can be useful in evaluating patients with unexplained dyspnea and fatigue. PMID- 12651054 TI - Left atrial volume as an index of left atrial size: a population-based study. AB - OBJECTIVES: We studied left atrial volume (LAV) as an index of atrial size. BACKGROUND: Increased left atrial dimension (LAD) measured by M-mode echocardiography is a risk factor for atrial fibrillation, stroke, and death. METHODS: A random sample of residents of Olmsted County, Minnesota, age > or =45 years (n = 2,042) underwent Doppler echocardiography with assessment of LAD and LAV. A subgroup of the population (n = 767) with no cardiovascular disease and normal systolic and diastolic function was used to develop reference ranges for LAD and LAV. In the total population, the prevalence of left atrial enlargement and the association between cardiovascular disease and left atrial size as determined by both indexes were assessed. RESULTS: In the normal subgroup, both indexes were associated with gender and body size, thus models controlling for body size were used to determine gender-specific reference ranges for LAD and LAV. In the total population, left atrial enlargement was common, with a prevalence of 18% (men) and 12% (women) using LAD/body surface area (BSA) and of 16% (men and women) using LAV/BSA. The agreement between the indexes was only fair (kappa = 0.53). Adjusting for age and gender, LAV/BSA was more strongly associated with the presence of cardiovascular diseases than LAD/BSA. CONCLUSIONS: We described a simple technique of measuring LAV, examined methods for indexing LAV, and described its normal range in a large, healthy reference cohort. Further, we find that in the community, left atrial enlargement is common and reflects the burden of cardiovascular disease. PMID- 12651055 TI - Insulin resistance and compensatory hyperinsulinemia: the key player between cigarette smoking and cardiovascular disease? AB - Hyperinsulinemia, dyslipidemia, and endothelial dysfunction are characteristic findings in insulin-resistant individuals, and all of these abnormalities have been identified as increasing cardiovascular disease (CVD) risk. Smokers tend to be relatively insulin resistant, hyperinsulinemic, and dyslipidemic, with evidence of endothelial dysfunction, as compared with nonsmokers, and recent epidemiologic data have suggested that CVD in smokers is primarily seen in those individuals who also have the characteristic findings of insulin resistance. Based on these observations, it is argued that insulin resistance and its consequences represent a major mechanistic link between cigarette smoking and CVD. It is also postulated that the enhanced CVD risk in smokers, resulting from hyperinsulinemia, abnormalities of lipoprotein metabolism, and endothelial dysfunction, will primarily be present in those smokers who are insulin resistant. As a corollary, it is suggested that CVD risk in individuals who cannot, or will not, stop smoking can be reduced by therapeutic efforts aimed at attenuating the adverse effects of insulin resistance and its consequences. PMID- 12651056 TI - Expression of lectin-like oxidized low-density lipoprotein receptors during ischemia-reperfusion and its role in determination of apoptosis and left ventricular dysfunction. AB - OBJECTIVES: The goal of this study was to determine the role of lectin-like oxidized low-density lipoprotein receptors (LOX-1), a recently identified oxidized low-density lipoprotein (ox-LDL) receptor, in ischemia-reperfusion injury to the heart. BACKGROUND: Reactive oxygen species (ROS) released during ischemia-reperfusion oxidize low-density lipoproteins; LOX-1 is upregulated by ox LDL and ROS, and is involved in cell injury. METHODS: Anesthetized rats were subjected to left coronary artery ligation for 60 min (n = 10, ischemia group), or ischemia followed by 60 min of reperfusion (n = 30, ischemia-reperfusion group). Rats in the latter group were treated with saline, the LOX-1 blocking antibody JXT21 (10 mg/kg), or nonspecific anti-goat immunoglobulin G (IgG) (10 mg/kg). Ten other rats underwent thoracotomy without coronary ligation (sham control). RESULTS: Ischemia-reperfusion was associated with an increase in LOX-1 expression, lipid peroxidation and apoptosis, a large infarct area, and a decrease in left ventricular function (all, p < 0.01 vs. sham control and ischemia alone groups). Treatment of rats with LOX-1 antibody prevented ischemia reperfusion-induced upregulation of LOX-1. Importantly, the LOX-1 antibody reduced apoptosis by 48%, lipid peroxidation by 39%, and myocardial infarct size by 45%, and improved left ventricular function (first derivative of pressure measured over time: -47% to -18%, p < 0.01). Nonspecific IgG had no effect. CONCLUSIONS: Lectin-like oxidized low-density lipoprotein receptors are upregulated during myocardial ischemia-reperfusion, and appear to be associated with apoptosis, necrosis, and left ventricular functional deterioration. PMID- 12651058 TI - Comparison of troponin T and creatine Kinase-MB fraction in evaluating cardiac patients postoperatively. PMID- 12651057 TI - Biodegradable gelatin hydrogel potentiates the angiogenic effect of fibroblast growth factor 4 plasmid in rabbit hindlimb ischemia. AB - OBJECTIVES: We investigated the potentiation of gene therapy using fibroblast growth factor 4 (FGF4)-gene by combining plasmid deoxyribonucleic acid (DNA) with biodegradable gelatin hydrogel (GHG). BACKGROUND: Virus vectors transfer genes efficiently but are biohazardous, whereas naked DNA is safer but less efficient. Deoxyribonucleic acid charges negatively; GHG has a positively charged structure and is biodegradable and implantable; FGF4 has an angiogenic ability. METHODS: The GHG-DNA complex was injected into the hindlimb muscle (63 mice and 55 rabbits). Gene degradation was evaluated by using (125)I-labeled GHG-DNA complex in mice. Transfection efficiency was evaluated with reverse-transcription nested polymerase chain reaction and X-Gal histostaining. The therapeutic effects of GHG FGF4-gene complex (GHG-FGF4) were evaluated in rabbits with hindlimb ischemia. RESULTS: Gelatin hydrogel maintained plasmid in its structure, extending gene degradation temporally until 28 days after intramuscular delivery, and improving transfection efficiency. Four weeks after gene transfer, hindlimb muscle necrosis was ameliorated more markedly in the GHG-FGF4 group than in the naked FGF4-gene and GHG-beta-galactosidase (control) groups (p < 0.05, Kruskal-Wallis test). Synchrotron radiation microangiography (spatial resolution, 20 microm) and flow determination with microspheres confirmed significant vascular responsiveness to adenosine administration in the GHG-FGF4 group, but not in the naked FGF4-gene and the control. CONCLUSIONS: The GHG-FGF4 complex promoted angiogenesis and blood flow regulation of the newly developed vessels possibly by extending gene degradation and improving transfection efficiency without the biohazard associated with viral vectors. PMID- 12651061 TI - Quantitative relationship between severity of pulmonary hypertension and LV diastolic function has been established. PMID- 12651063 TI - Detrimental effects of late artery opening. PMID- 12651064 TI - Specificity of noninvasive pacemaker stress echocardiography in diagnosis of coronary artery disease. PMID- 12651066 TI - Molecular cloning of IBP, a SWAP-70 homologous GEF, which is highly expressed in the immune system. AB - Rho GTPases play a fundamental role in a variety of biological processes ranging from the reorganization of the actin cytoskeleton to the regulation of cell proliferation. The activation of Rho GTPases is regulated by guanine nucleotide exchange factors (GEFs) belonging to the Dbl family of proteins. The hallmark of this large family of GEFs is the presence of a tandem DH-PH module in which a pleckstrin-homology (PH) domain is located at the C-terminus of a Dbl-homology (DH) domain. Recent studies have demonstrated that SWAP-70 constitutes a novel class of Rac-GEF, in which the PH domain is located at the N-terminus, rather than the C terminus, of the DH domain. Here we report the molecular cloning of human IBP (IRF-4 binding protein), a new member of this novel family of GEFs. The IBP gene maps to human chromosome 6p21.31 centromeric to the MHC locus. Isolation of the murine IBP cDNA reveals a very high degree of homology with the human IBP cDNA suggesting that IBP is evolutionarily conserved. The 5' portion of the murine IBP cDNA is furthermore identical to the Def-6 cDNA fragment, which was identified in the course of a search for genes differentially expressed in the murine hematopoietic system. IBP is broadly expressed in the immune system and can be detected in both T and B cell compartments in contrast to SWAP-70 whose expression is primarily restricted to B cells. Taken together these findings indicate that IBP is a novel type of GEF, which participates in the activation of Rho GTPases in lymphoid tissues. PMID- 12651067 TI - Genetic control of interleukin-4-induced activation of the human signal transducer and activator of transcription 6 signaling pathway. AB - The interleukin (IL)-4-induced Stat6 signaling pathway is active in a variety of cell types, including immune cells and cancer cells, and plays an important role in the regulation of gene expression, such as CD23 and major histocompatibility complex class II. Using a semiquantitative gel shift assay in which nuclear Stat6 activities were scored, three Stat6 activation phenotypes were defined as Stat6(high) (intense banding), Stat6(low) (medium intensity banding), and Stat6(null) (very low to no discernible banding). These Stat6 phenotypes correlated well with levels of CD23 expression, but not with those of human leukocyte antigen-DR cell-surface display. Pedigree analyses revealed a Mendelian inheritance pattern that can be explained by two STAT6 Pathway (STAT6P) activation genotypes, which we term A and a, where STAT6P*A determines an active Stat6 signaling and STAT6P*a determines an inactive Stat6 signaling, with incomplete dominance. Total Stat6 protein levels failed to correlate with the above Stat6 phenotypes allowing us to propose that IL-4-induced Stat6 signaling is a polygenic quantitative trait regulated by a collection of several contributing genetic loci that functionally interact. The Stat6(null) phenotype may result from a defect in Stat6 signaling, which has important implications with respect to the pathogenesis of cancer and Th1/Th2 cytokine imbalance in autoimmune diseases in general. PMID- 12651068 TI - Light stimulates IFNgamma-mediated intercellular adhesion molecule-1 upregulation of cancer cells. AB - Intercellular adhesion molecule-1 (ICAM-1) works as one of the ligands for activating the killing activity of natural killer (NK) cells and cancer specific cytotoxic T lymphocytes (CTL). Expression of ICAM-1 enhances lymphocyte adhesion to the cancer cells in vivo. Cancer cell lines express significantly lower level of ICAM-1 than that of normal epithelium or benign cells. Overexpression of LIGHT (LIGHT: homologous to lymphotoxins, indicating inducible expression, and competes with herpes simplex virus glycoprotein D for herpes virus entry mediator [HVEM/TR2]) in MDA-MB-231 human breast cancer cells was observed to suppress tumor growth in vivo. In order to elucidate the mechanisms how LIGHT overexpression could trigger tumor suppression, the expression level of a panel of cell surface makers CD54, CD56, CD95, and CD119 was investigated in a group of cancer cells. Flow cytometry analysis results demonstrate that LIGHT gene expression in cancer cells can greatly increase ICAM-1 expression level, IFNgamma alone can stimulate cancer cells to express ICAM-1, which can be highly augmented by LIGHT in a dose-dependent manner. This upregulation of ICAM-1 expression is not only at ICAM-1 protein trafficking level on cell surface as demonstrated by flow cytometry analysis, but also at ICAM-1 total protein level as confirmed by Western blot. There is no difference of expression level among these cancer cell lines for the other three cell surface markers: CD56, CD95 (Fas), and CD119. It was confirmed that LIGHT enhancement upregulation of ICAM-1 expression is at least STAT1 and JAK1 dependent by using STAT1-deficient U3A and JAK1-deficient E2A4 cells. These findings suggest that LIGHT-induced inhibition of tumor growth is highly correlated with its upregulation of ICAM-1 expression. PMID- 12651069 TI - Umbilical cord blood dendritic cells are a rich source of soluble HLA-DR: synergistic effect of exosomes and dendritic cells on autologous or allogeneic T Cell proliferation. AB - In this study, we compared soluble HLA-DR (sHLA-DR) production in the culture supernatants of various cell sources [T and B cells, monocytes and dendritic cells (DCs) either from adult peripheral blood (PB) or umbilical cord blood (UCB)]. DCs produced the highest amount of sHLA-DR molecules as compared to other cell sources, with UCB DCs producing the highest amount. Different kinetics of sHLA-DR production were found between immature and mature UCB DCs (mDC, iDC) (derived either from CD34(+) or CD14(+) cells). Maximum production of sHLA-DR was observed in 72-hour culture supernatants of both CD34- and CD14-derived mDCs, whereas it peaked in the 24-hour culture supernatants from iDC. sHLA-DR molecules were pelleted after sequential centrifugation from UCB CD34(+) DCs and were found to contain both 36 kD alpha-chain and 29 kD beta-chain of HLA-DR, CD86, and Fas molecules. These sHLA-DR containing vesicles/exosomes alone evoked weak proliferative responses from autologous and allogeneic T cells, but the immune response was significantly increased when vesicles/exosomes were presented with DCs. PMID- 12651070 TI - Relative dominance of HLA-B*07 restricted CD8+ T-lymphocyte immune responses to human cytomegalovirus pp65 in persons sharing HLA-A*02 and HLA-B*07 alleles. AB - CD8(+) T-cell responses to three human cytomegalovirus (CMV) pp65 epitopes were studied in panels of healthy seropositive HLA-A*02/HLA-B*07 individuals, and HLA A*02 donors mismatched for HLA-B*07. The majority of the latter had significant responses to a HLA-A*02-restricted epitope within the CMV pp65 antigen. By contrast, the strongest responses to CMV in the first group were to HLA-B*07 restricted epitopes. Similar immunodominance of HLA-B*07 over HLA-A*02 was found in two immunocompromised HIV-infected HLA-A*02/HLA-B*07 patients, and in the reconstituting immune system of three stem cell transplant recipients. In vitro stimulation of peripheral blood mononuclear cells (PBMC) from two immunocompetent HLA-A*02/HLA-B*07 individuals indicated that cytotoxic T lymphocyte (CTL) precursors specific for both HLA-A*02 and HLA-B*07 restricted epitopes were present and could be expanded by stimulation with the cognate peptides. However, if stimulation was performed by antigen presenting cells infected with recombinant vaccinia expressing full-length native pp65, only HLA-B*07 epitope specific cells were seen. In one patient the HLA-B*07 dominance was partially broken by using recombinant vaccinia expressing ubiquitinated pp65, suggesting that enhanced protein processing can reveal weaker immune responses. Our results indicate that CMV-specific cellular immune responses restricted by HLA-B*07 dominate those restricted by HLA-A*02 in both immunocompetent and immunocompromised individuals. This may have significant consequences for the design of epitope-specific vaccines. PMID- 12651072 TI - Distribution of LILRA3 (ILT6/LIR4) deletion in psoriatic patients and healthy controls. AB - The leukocyte immunoglobulinlike receptor (LILRA3; ILT6) gene is localized on human chromosome 19 in the region 19q13.4, in the leukocyte receptor complex that encodes leukocyte receptors LILR (ILT/LIR), killer cell immunoglobulinlike receptors (KIR), LAIR, Fc IgA receptor, and others. The biologic role of the LILRA3 molecule and the nature of its ligand are not known. Comparison of LILRA3 gene sequence with those of other LILRs suggests LILRA3 is a soluble molecule. If LILRA3 binds human leukocyte antigen (HLA) class I molecules like other LILRs whose ligands are known, then it might block recognition of HLA by these receptors, influencing immune response and susceptibility to HLA class I associated disease. A deletion of LILRA3 gene was found in a minority of British population. We typed 108 healthy individuals from the Low Silesia region and 103 patients diagnosed with psoriasis vulgaris (a disease associated with HLA class I antigen, HLA-Cw6) for LILRA3 to examine whether LILRA3 deletion was distributed differently in patients affected with the disease. No differences in frequencies of the LILRA3 deletion were found between controls and patients or between HLA Cw6(+) and HLA-Cw6(-) controls or patients, suggesting that LILRA3 has no role in psoriasis. PMID- 12651071 TI - Influence of cytokine and mannose binding protein gene polymorphisms on human T cell leukemia virus type I (hTLV-I) provirus load in HTLV-I asymptomatic carriers. AB - Human T-cell leukemia virus type I (HTLV-I) provirus load differs more than 100 fold among carriers and a high provirus load in the peripheral blood mononuclear cells (PBMCs) is regarded as a risk factor for both preleukemic states and inflammatory diseases including HTLV-I-associated myelopathy (HAM). We examined polymorphisms in the genes for tumor necrosis factor (TNF), TNF receptor type 1 and 2, lymphotoxin (LT)-alpha, interleukin (IL)-1beta, IL-6, IL-10, monocyte chemoattractant protein (MCP)-1, and mannose binding protein (ManBP) in 143 HTLV I carriers whether these polymorphisms affect the provirus load in the PBMCs of carriers. No significant association was observed between these polymorphisms and the provirus load. Homozygotes for a ManBP-variant allele, however, showed a tendency for the decreased number of provirus load. When combined, the data on the alleles of LT-alpha and MCP-1, HTLV-I carriers having high producer alleles of both genes showed a trend for increased provirus load. These data suggest that inflammation or an active immune response may induce an increased amount of HTLV I-infected T cells, leading to a high provirus load. PMID- 12651073 TI - HLA class II antigens associated with lupus nephritis in Italian SLE patients. AB - Human leukocyte antigen DR2 (HLA-DR2), namely the allelic variant HLA-DR15, have been associated with lupus nephritis (LN) in Caucasians. The study investigated the relationships between HLA class II alleles and lupus nephritis in Italian patients. Two hundred forty-four patients fulfilling the American Rheumatism Association criteria for systemic lupus erythematosus (SLE) were typed for HLA DRB1*, -DQA1*, -DQB1*, and -DPB1* alleles by polymerase chain reaction-sequence specific oligonucleotide and polymears chain reaction-single-strand polymorphism; 71 patients had renal damage assessed by renal biopsy. Glomerulonephritis was classified using WHO criteria. Significance was tested by X(2) on 2x2 tables. HLA DQA1*0101 was strongly associated with LN (OR = 2.72 [1.43-5.19]; p = 0.002), whereas HLA-DRB1*1501 was only marginally associated (OR = 1.94 [0.88-4.26]; p = not significant). HLA-DQA1*0102 demonstrated a significant protective effect (OR = 0.31 [0.14-0.86]; p = 0.002). On analyzing the distribution of HLA-DRB1*1501 bearing haplotypes in our SLE patients we found that the HLA-DRB1*1501 greatly enhanced the risk of developing LN conferred by the DQA1*0101 allele (OR = 65.96 [9.35-1326.25]), whereas DQA1*0102 suppressed the nephritogenic effect of DRB1*1501. At renal biopsy, 80% of DRB1*15 positive patients were classified as having class IV LN with the remaining 20% having class III LN. The figures were 19% and 21%, respectively, among the HLA-DR15 negative patients. In the Italian population HLA-DQA and HLA-DR alleles interact in conferring susceptibility to or protection against lupus nephritis, the diffuse proliferative glomerulonephritis (i.e., the most severe form of nephritis) is associated with the HLA-DR15 bearing haplotypes. PMID- 12651074 TI - HLA types in celiac disease patients not carrying the DQA1*05-DQB1*02 (DQ2) heterodimer: results from the European Genetics Cluster on Celiac Disease. AB - Genetic susceptibility to celiac disease is strongly associated with HLA-DQA1*05 DQB1*02 (DQ2) and HLA-DQA1*03-DQB1*0302 (DQ8). Study of the HLA associations in patients not carrying these heterodimers has been limited by the rarity of such patients. This European collaboration has provided a unique opportunity to study a large series of such patients. From 1008 European coeliacs, 61 were identified who neither carry the DQ2 nor DQ8 heterodimers. Fifty seven of these encoded half of the DQ2 heterodimer. The remaining 4 patients had a variety of clinical presentations. Three of them carried the DQA1*01-DQB*05 haplotype as did 20/61 of those carrying neither DQ2 nor DQ8. This may implicate a role of the DQA1*01 DQB*05 haplotype. None of these four patients carried the DQB1*06 allele that has previously been reported in this sub-group of patients. Of the 16 DQ2 heterodimer negative patients without DRB1*04 or DRB1*07 haplotypes, it was inferred that none encoded the previously implicated DRB4 gene as none had a DRB1*09 haplotype. These results underline the primary importance of HLA-DQ alleles in susceptibility to celiac disease, and the extreme rarity of celiac patients carrying neither the DQ2 or DQ8 heterodimers nor one half of the DQ2 heterodimer alone. PMID- 12651075 TI - Evidence for novel DRB1*15 allele association among clinically definite multiple sclerosis patients from Mumbai, India. AB - Multiple sclerosis (MS) is a clinically heterogeneous demylinating disease and an important cause of acquired neurologic disability. MS has been reported from different regions of India and its infrequency has been attributed to have genetic implications. Further, a high incidence of MS and its human leukocyte antigen B12 (HLA-B12) associations have been reported among highly inbred Parsi population from Mumbai. However, consistent HLA associations have not been reported from India. We analyzed the HLA-B, -Cw, and -DRB1 allele associations among 23 clinically definite Western Indian non-Parsi MS patients and compared them with 146 ethnically matched clinically normal individuals. HLA serologic (A, B, and Cw) as well as molecular (DRB1) typing methodology was followed. The study revealed a significant increase of HLA-A11 (24% vs. 13%; OR = 2.6; EF = 0.14; 95%CI = 1.1-3.05), B16 (4.3% vs 0.3%; OR = 13.8; EF = 0.03; 95% CI = 1.19 134.44), Cw7 (15.2% vs 3.7%; OR = 5.46; EF = 0.12; 95% CI = 0.944-17.86), and DRB1*15 (21.7% vs 2.2%; OR = 16.15; EF = 0.19; 95% CI = 1.33-68.64). Further molecular subtyping of HLA-DRB1*15 among the patients revealed two novel alleles, DRB1*1506 (20%) and DRB1*1508 (30%), along with the commonly reported DRB1*1501 (50%) for the first time in MS patients that were hitherto unidentified from other parts of India and world as well. This study reveals that there is a complexity of the genetic susceptibility to MS in different populations studied and reported. PMID- 12651076 TI - Efficacy and tolerability of levetiracetam during 1-year follow-up in patients with refractory epilepsy. AB - PROBLEM: Levetiracetam (LEV) is a new antiepileptic drug shown to be effective for the treatment of partial seizures in pivotal clinical trials. We investigated the long-term efficacy and tolerability of LEV as add-on therapy, regardless of seizure type, especially in persons who would not be eligible for clinical trials due to factors such as mental retardation and concomitant psychiatric disorders. METHODS: Ninety-eight patients participated and were followed for 1 year. Demographic data, seizure frequency, and side effects were recorded at baseline and during the 1-year follow-up. The first 35 patients were given LEV at a starting dose of 500 mg b.i.d. with weekly increments of 1000 mg (fast titration). The other patients were given LEV with a starting dose of 250 mg b.i.d. with weekly increments of 250 mg (slow titration). RESULTS: Fourteen patients were completely seizure free after titration to effective dose and 57 were responders with >50% seizure reduction for the first year. In the group with generalized seizures, 1 out of 19 became seizure free, but 8 patients had >50% decrease. Average dose at 1 year was 1900 mg (+/-900). Seventeen of 38 discontinuations were due to adverse effects and 21 were due to lack of efficacy. With fast titration, 15 out of 35 (43%) experienced tiredness during the first 12 weeks, and with slower titration 20 of 63 (32%) experienced tiredness. The difference was not statistically significant. Four out of the five patients who discontinued due to behavioral adverse events (mainly irritability) previously had behavioral problems and/or mental retardation. One patient discontinued due to psychosis. CONCLUSIONS: Levetiracetam appears to be well tolerated in patients with severe epilepsy and shows efficacy in a long-term follow-up. Behavioral adverse events were noted in a small number of patients and occurred mainly in patients who had a history of behavioral disturbance or were mentally retarded. These data from an open population are consistent with the findings of clinical trials. PMID- 12651077 TI - Clinical experience of marketed Levetiracetam in an epilepsy clinic-a one year follow up study. AB - Levetiracetam is a new anti-convulsant with impressive pivotal trial credentials. We examined its effectiveness in refractory clinic patients with epilepsy with a year's follow up. Six months after initiation 32% of the patients were seizure free, and 26% at one year. By the end of the 12 months follow up 77% of patients were still taking the drug, having gained benefit from it: 23% had dropped out due to intolerable side effects, seizure increase or lack of efficacy. There is evidence that the drug is broad spectrum and as effective in primary generalised epilepsy as in partial onset epilepsy. Our audit of its use and effectiveness has led us to position it as our first choice add-on drug if the initial monotherapy drug fails. PMID- 12651078 TI - An open-label study of levetiracetam at individualised doses between 1000 and 3000 mg day(-1) in adult patients with refractory epilepsy. AB - BACKGROUND: The novel antiepileptic drug (AED) levetiracetam (LEV, Keppra) is indicated as adjunctive therapy for partial epilepsy. The primary aim of this study was to measure the safety and tolerability of LEV individualised dosing in a heterogeneous refractory epilepsy population. METHODS: LEV was evaluated in a 10- to 16-week open-label, multicentre study in adult patients with epilepsy refractory to previous treatment with at least two AEDs. Individualised LEV doses up to 3000 mg x day(-1) were determined in an initial up-titration phase, and optimal doses were administered as adjunctive treatment during an 8- to 10-week evaluation period. Concomitant AEDs and their doses could not be changed during the study. Safety and tolerability were monitored by expression of adverse events as well as by retention rate. The effect of LEV on concomitant AED concentration was also studied. Efficacy was assessed using global clinical evaluation (GCE) scores, seizure frequency, and >or=50% responder rate. RESULTS: LEV therapy was initiated in 219 patients; 183 had localisation-related epilepsy and 37 had generalised epilepsy. In one patient, epileptic syndrome was defined as both localisation-related and generalised. About 81.7% (179/219) continued and completed treatment throughout the study, and 79% (172/219) chose to continue LEV in a follow-up study. The most common adverse events were asthenia, dizziness, and somnolence. Most adverse events occurred during up-titration. LEV treatment did not alter the concentration of concomitant AEDs. LEV improved GCE scores in 79.5% (152/191) of patients. LEV reduced the median total seizure frequency of all patients from a median of 2.25 seizures per week at baseline (n=219) to 1.10 seizures per week during the evaluation period (n=191 patients with at least one seizure count during evaluation). The >or=50% responder rate was 48.2% for all seizure types, 49.4% for partial-onset, and 51.4% for generalised-onset seizures. Throughout the evaluation period (i.e. from the start of the evaluation period until completion or early discontinuation), 26/191 (13.6%) had a 100% reduction in total seizure frequency, while in a follow-up study, 10.5% (18/172) were seizure-free for at least 6 months and 6.4% (11/172) were seizure-free for at least 1 year. CONCLUSION: LEV was well tolerated, as evidenced by limited adverse event reporting and the high retention rate, and appeared effective in both generalised and partial epilepsy. PMID- 12651079 TI - Levetiracetam monotherapy for primary generalised epilepsy. AB - PURPOSE: To evaluate the efficacy of levetiracetam in cases of refractory primary generalised epilepsy. METHODS: Three patients with refractory primary generalised epilepsy were treated with levetiracetam monotherapy; one with absence seizures, myoclonic jerks and generalised tonic-clonic (GTC) seizures one with myoclonic jerks and GTC seizures, and one with only GTC seizures. All three patients had generalised spike wave on the EEG and had failed at least three antiepileptic drugs (AEDs) before trying levetiracetam. RESULTS: All three patients tolerated levetiracetam well and became seizure free for at least 6 months. Therapeutic doses of levetiracetam ranged from 1250 to 3000 mg/day. CONCLUSION: Levetiracetam, a new AED with a novel mechanism(s) of action, should be considered for patients with refractory primary generalised epilepsy. PMID- 12651080 TI - Levetiracetam monotherapy for newly diagnosed epilepsy patients. AB - We retrospectively reviewed the charts of all of our patients with a history of partial seizures, with and without secondarily generalisation, who received levetiracetam (LEV; Keppra) for treatment of their seizures during the years 2000 2002. Forty-five patients were identified, 13 of whom began LEV as first line therapy. Eleven patients continued on LEV for at least 6 months; six of whom became seizure free and five had >50% reduction in their seizures. The remaining two patients discontinued LEV because of adverse effects. LEV monotherapy can be effective and well tolerated in adults with new onset seizures. A prospective, large, double-blind monotherapy study for newly diagnosed patients is needed to confirm this finding. PMID- 12651081 TI - Levetiracetam for benign epilepsy of childhood with centrotemporal spikes-three cases. AB - Benign epilepsy with centrotemporal spikes (BECTS), also known as benign rolandic epilepsy (BRE) of childhood represents 15% of all childhood epilepsies [Handbook of Epilepsy Treatment (2000)]. A majority of these patients do no require treatment; however, in those cases where treatment is justified, the most efficacious medication with a benign safety profile should be selected. We present three clinical cases of otherwise healthy children with BECTS who were treated only with levetiracetam. All three of these children remain seizure-free and are experiencing no reported side effects. PMID- 12651082 TI - Surgery for frontal lobe epilepsy. PMID- 12651083 TI - Epilepsy surgery, delays and referral patterns-are all your epilepsy patients controlled? AB - RATIONALE: Epilepsy surgery is a standard of care in the treatment of medically intractable epilepsy, but is underutilised. We describe the results of epilepsy surgery and the referral patterns at a referral epilepsy programme. METHODS: We reviewed the outcome of epilepsy surgery performed at the University of South Florida and Tampa General Hospital epilepsy programme for the years 2000 and 2001. The typical presurgical evaluation included clinical evaluation, EEG-video monitoring, MRI with dedicated epilepsy protocol, PET, SPECT, neuropsychological testing and Wada testing. We used the Engel outcome classification, and focused on the referral information to determine how and when in the course of their illness patients arrive at a referral epilepsy centre. RESULTS: In the 2-year period (2000-2001), a total of 36 epilepsy surgeries were performed. Twenty-nine temporal lobectomies, six extratemporal resections and one corpus callosotomy. Ages varied from 17 to 65 years. Overall results were: 30 (83%) seizure-free [class I], 5 (17%) rare seizures or almost seizure-free [class II] and 1 no improvement. Of the 29 temporal lobectomies, 27 (93%) are completely seizure-free [class I] and 2 (7%) are >90% improved [class II]. Duration of seizures before being seen at the epilepsy centre averaged 18 years (range 2-58 years). Twenty two (61%) were sent by their neurologists, while 14 (39%) came self-referred without having discussed surgery with their neurologists. Five (14%) were specifically advised by their neurologist to not consider surgery. Two had participated in clinical trials of antiepileptic drugs (AEDs) before being seen at the epilepsy centre. CONCLUSIONS: Epilepsy surgery has high efficacy and very low morbidity. Yet, there continues to be a long delay in the referral of patients to the epilepsy centre, suggesting that surgery for epilepsy is underutilised. PMID- 12651084 TI - Risk factors for childhood epilepsy: a case-control study from Irbid, Jordan. AB - OBJECTIVE: The goal of this case-control study is to identify the significance of certain risk factors for epilepsy in a population of epileptic children in Northern Jordan. The risk factors examined are febrile convulsions, head trauma, central nervous system infections, abnormal perinatal history, family history and parental consanguinity. METHODOLOGY: We designed a case-control study for patients attending the outpatient neurology clinic of Princess Rahma Teaching Hospital in Irbid, Jordan during a 7-month period. Controls were selected, matched for age and sex, from a group of non-epileptic patients attending the general paediatrics outpatient clinic in the same hospital and during the same period. Data about the investigated risk factors were obtained by personal interview and review of the medical records and were analysed statistically for significance. RESULTS: The total number of participants was 200 patients and controls each. History of febrile convulsions, head trauma, abnormal perinatal history and family history showed a statistically significant increase risk for developing epilepsy. Central nervous system infections and parental consanguinity did not add to the risk of developing epilepsy. CONCLUSION: Positive family history for epilepsy, head trauma, febrile convulsions and abnormal perinatal history were shown to have a statistically significant association with epilepsy in patients attending Princess Rahma Teaching Hospital in Northern Jordan. Although consanguinity is widely practised in Jordan, it appears that it does not increase the risk of epilepsy probably due to the small contribution of monogenic recessive epilepsies to the population with epilepsy. PMID- 12651085 TI - The use of the ketogenic diet in a patient with subacute sclerosing panencephalitis. AB - OBJECTIVE: To report on the effects of the ketogenic diet on a 9-year-old boy with myoclonic jerks due to subacute sclerosing panencephalitis (SSPE). METHODS: A 9-year-old boy presented with progressively worsening myoclonus unresponsive to valproic acid and clonazepam. He was started on the ketogenic diet maintaining urine ketones at greater than 80 mg x dl(-1). RESULTS: Within 2 weeks of dietary initiation, myoclonic jerks stopped. Four weeks later he developed cognitive slowing. Results of electroencephalogram and cerebrospinal fluid analysis were consistent with SSPE. Three months after ketogenic diet initiation, myoclonic jerks reappeared and were refractory to treatment. CONCLUSION: The ketogenic diet may be useful in controlling, even temporarily, the myoclonic jerks of SSPE. PMID- 12651086 TI - Somatisation indexes as differential factors in psychogenic pseudoepileptic and epileptic seizures. AB - One of the postulated factors responsible for psychogenic epileptic seizures is somatisation. The purpose of this study was to analyse differences in the levels of somatisation manifested in the Minnesota Multiphasic Personality Inventory (MMPI) somatisation scales. Subjects were divided into three groups on the basis of a neurological examination and long-term video-monitoring. Group One (N=66, 55F; 11M) had only psychogenic pseudoepileptic seizures. Group Two (N=42, 32F; 10M) had both epileptic and psychogenic pseudoepileptic seizures. Group Three (N=36, 28F; 8M) had only epileptic seizures and served as the control group. Patients in all three groups were given the MMPI. Somatisation indexes in the three groups were compared. Significant between-group differences were obtained for the following somatisation indexes (mean scores): Hypochondriasis (P<0.001), Somatic Complaints (P<0.001), Organic Symptoms (P<0.015) and Poor Health (P<0.05). No significant differences were found for Physical-Somatic Complaints. Psychological evaluation of the differences in the levels of somatisation in these groups may help us to gain a better understanding of, and discrimination between, patients with psychogenic epileptic seizures, mixed seizures and epileptic seizures only. PMID- 12651087 TI - Automatisms with preserved responsiveness and ictal aphasia: contradictory lateralising signs during a dominant temporal lobe seizure. AB - The 25-year-old right-handed woman suffering from temporal lobe epilepsy (TLE) was referred to our centre for presurgical evaluation. MRI showed a right-sided hippocampal sclerosis. During video-EEG-recorded seizures, abdominal aura was followed by oral automatisms, during which she was completely reactive to external stimuli, although she was unable to speak. Ictal EEG showed right temporal seizure pattern, without contralateral propagation. She had abnormal speech postictally. Speech-activated functional transcranial Doppler sonography revealed right-sided speech dominance. She has become seizure free after a right sided amygdalo-hippocampectomy. In our patient, contradictory clinical ictal lateralising signs (automatisms with preserved responsiveness vs. ictal and postictal dysphasia) occurred during right-, speech-dominant-sided seizures. This is the first report when automatisms with preserved consciousness occurred during a seizure originating and involving the speech-dominant hemisphere. PMID- 12651088 TI - Persistent hyperinsulinemic hypoglycaemia followed as benign infantile convulsion. AB - An 18-month-old boy developed seizures at 3 months of age. He developed normally and, his EEG and brain CT revealed no abnormal findings. The blood sugar level was normal at that time, thus he was diagnosed as having benign infantile convulsion. At 7 months of age seizures reappeared, and hypoglycaemia associated with hyperinsulinism was observed during the seizures. With conservative therapy his blood sugar level was well controlled and he had no further seizures. Hypoglycaemic seizures are sometimes misdiagnosed as epilepsy. We have to pay attention to hyperinsulinemic hypoglycaemia when we see seizures with normal EEG even in infants. PMID- 12651094 TI - An overview of the Notch signalling pathway. AB - The Notch receptor plays a key role in modulating cell fate decisions throughout the development of invertebrate and vertebrate species. Four Notch homologues have been identified in the human genome and aberrant Notch function has been associated with a number of human diseases. An intriguing pathway of Notch signalling has been elucidated involving multiple proteolytic steps and this pathway offers several targets for potential therapeutic intervention. While a consensus model has emerged, in the details there is much that is contentious. This review will primarily focus on our current understanding of the proteolytic events involved in generating and regulating Notch signalling. PMID- 12651095 TI - A Notch so simple influence on T cell development. AB - T cell precursors undergo a series of developmental choices that progressively narrow their ability to give rise to different cell lineages. Evidence accumulated in the last few years suggests that Notch occupies a central place among the signal transduction pathways that regulate many of these choices, including the T/B, alphabeta/gammadelta and CD4/CD8 lineage decisions. Nevertheless the mechanisms by which Notch exerts its effects are not well understood, and in some cases the physiologic role is unclear. In this review we try to present succinctly the experiments and highlight the areas of controversy. PMID- 12651096 TI - Notch signalling in the regulation of peripheral T-cell function. AB - The Notch signalling pathway plays a highly-conserved role in regulating the cellular differentiation and proliferation events that characterise pattern formation in the embryo. As cells in the embryo respond to environmental signals, similarly T-cells in the peripheral immune system must monitor their environment for antigens and respond accordingly by entering one of several potential differentiation pathways. Recent studies have identified a role for the Notch pathway in regulating the responses of T-cells in the periphery. In this review, we discuss these findings in the context of the Notch signalling pathway's role as an orchestrator of cellular differentiation, and propose a central role for Notch as a regulator of immune system function. PMID- 12651097 TI - Notch signalling in B cells. AB - Notch signalling is likely to regulate multiple aspects of lymphoid development and function. During T cell development, Notch signalling is required for commitment of the earliest progenitor, and may also function during other developmental stages. T cell commitment from a common lymphoid progenitor occurs at the expense of B cell development, suggesting that Notch signalling inhibits the earliest stage of B lymphopoiesis. In contrast, recent evidence suggests that Notch promotes the development of marginal zone lymphocytes. Not only is Notch required for later stages of B cell development, but several viral proteins appear to utilize Notch signalling in B cells to mediate their functions. In this review, we will focus on potential roles of Notch signalling in B lymphopoiesis and also consider how viral proteins may utilize Notch signalling in B cells. PMID- 12651098 TI - Notch signalling in hematopoiesis. AB - The Notch pathway is a widely utilized, evolutionarily conserved regulatory system that plays a central role in the fate decisions of multipotent precursor cells. Notch often acts by inhibiting differentiation along a particular pathway while permitting or promoting self-renewal or differentiation along alternative pathways. Haematopoietic cells and stromal cells express Notch receptors and their ligands, and Notch signalling affects the survival, proliferation, and fate choices of precursors at various stages of haematopoietic development, including whether haematopoietic stem cells self-renew or differentiate, common lymphoid precursors undergo T or B cell differentiation, or monocytes differentiate into macrophage or dendritic cells. These findings suggest that the Notch pathway plays a fundamental role in regulating haematopoietic development. PMID- 12651099 TI - Notch signalling at the crossroads of T cell development and leukemogenesis. AB - Members of the Notch family (e.g. Notch1 and Notch3) have been recently described to play a critical role in T cell development and their constitutive activation has been related to T cell leukaemia in both animal models and human disease. Nevertheless, whether they act as redundant molecules, by affecting the same molecular mechanisms, or play distinct roles in T cell differentiation and/or leukemogenesis is not clear. Altered Notch signalling impairs the developmentally regulated interplay between pre-TCR signalling, NFkappaB and E2A activities, thus identifying the crucial role of Notch receptors at the cross-roads of disrupted lymphoid differentiation and neoplastic transformation. PMID- 12651100 TI - Practical considerations in refolding proteins from inclusion bodies. AB - Refolding of proteins from inclusion bodies is affected by several factors, including solubilization of inclusion bodies by denaturants, removal of the denaturant, and assistance of refolding by small molecule additives. We will review key parameters associated with (1) conformation of the protein solubilized from inclusion bodies, (2) change in conformation and flexibility or solubility of proteins during refolding upon reduction of denaturant concentration, and (3) the effect of small molecule additives on refolding and aggregation of the proteins. PMID- 12651101 TI - Alocasin, an anti-fungal protein from rhizomes of the giant taro Alocasia macrorrhiza. AB - An anti-fungal protein designated alocasin was isolated from the rhizomes of the giant taro Alocasia macrorrhiza. The isolation protocol involved ion exchange chromatography on diethylaminoethyl (DEAE)-cellulose, ion exchange chromatography on sulfopropyl (SP)-Sepharose, and gel filtration on Superdex 75. Alocasin, which was unadsorbed on DEAE-cellulose and SP-Sepharose, possessed the N-terminal sequence APEGEV, which exhibited some similarity to that of the miraculin-like anti-fungal protein from Pisum sativum legumes. It demonstrated a molecular mass of 11kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration, and displayed anti-fungal activity against Botrytis cinerea. Alocasin reduced the activity of HIV-1 reverse transcriptase. It exhibited weak hemagglutinating activity, only at a concentration of 1mg/ml. PMID- 12651102 TI - Expression, purification, and PC1-mediated processing of human proglucagon, glicentin, and major proglucagon fragment. AB - To examine the cleavage specificity of different members of the furin/propeptide convertase (PC) family of enzymes, we have selected proglucagon (PG) as a model substrate. PG was selected because it is subject to differential processing in vivo. PG is thought to be cleaved initially at an interdomain site to produce glicentin and the major proglucagon fragment (MPGF). These intermediates are subsequently cleaved, most likely by the convertases PC2 and PC1, respectively. To determine the exact sites within PG that are cleaved by PC1 and PC2, we attempted to produce milligram quantities of human PG, glicentin, and MPGF for use in an in vitro conversion assay. A methionine residue was added to the N terminus of each protein to initiate translation. Purification was achieved using cation exchange and reversed-phase chromatography, and the integrity of the methionylated proteins was confirmed by both electrospray ionization-mass spectrometry and amino acid analysis. The combined expression and purification scheme is fast, efficient, and results in milligram quantities of > or =95% pure proglucagon, > or =95% pure MPGF, and > or =93% pure glicentin. These prohormones are cleaved by PC1 to produce product peptides consistent with the processing of PG observed in vivo, and should therefore be suitable for further analysis of the post-translational processing of PG. PMID- 12651103 TI - The relative ligand binding preference of the murine ileal lipid binding protein. AB - The ileal lipid binding protein (ILBP), a member of the intracellular lipid binding protein family, is a 14-kDa protein that has bile and fatty acids as possible physiological ligands. The ligand binding specificity of this protein is not well characterized. Therefore, we studied the lipid binding activity of purified recombinant murine ILBP (mILBP) in vitro. These studies demonstrated by direct analysis the interaction of mILBP with naturally occurring bile and fatty acids. The rank order of binding preference for fatty acids, or unconjugated and conjugated bile acids, was assessed. Among fatty acids, mILBP preferred species that had longer chain length and increased saturation, similar to other members of the intracellular lipid binding protein family. Among the bile acids, mILBP showed the greatest preference for conjugated species that contained a doubly hydroxylated steroid moiety. The results demonstrate that mILBP exhibits a preference for certain species of bile and fatty acids. PMID- 12651104 TI - Expression, refolding, and in vitro activation of a recombinant snake venom pro metalloprotease. AB - Metalloproteases comprise a family of Zn(2+)-endopeptidases that degrade most components of the extracellular matrix. Snake venoms are rich sources of metalloproteases, which also digest fibrinogen as well as fibrin, and in some cases, induce hemorrhage. A few low-molecular weight snake venom metalloproteases (svMPs) have been described as being devoid of hemorrhagic activity, but they have strong direct-acting fibrinolytic activity. This property could be very helpful in thrombosis therapy. ACLF is a fibrinolytic, non-hemorrhagic metalloprotease from the venom of the North American snake Agkistrodon contortrix laticinctus. We have developed an expression system for production of a recombinant pro-ACLF from a clone (ACLPREF) isolated from a venom gland cDNA library. The coding region including both the pro-enzyme domain and the mature protein domain was amplified by PCR and subcloned into the pET28a vector and the new plasmid was used to transform BL21(DE3) Escherichia coli cells. Culture of the transformants at 37 degrees C led to the overexpression of an insoluble 48kDa protein after induction with 1.0mM IPTG. The expressed protein was recovered from inclusion bodies with 6M buffered urea and purified by affinity chromatography under denaturing conditions. After dithiothreitol treatment, protein refolding was performed by gradual removal of the denaturing agent by dialysis. The pro enzyme underwent auto-activation during refolding and it was active on fibrinogen and on a synthetic substrate. To control the activation step, the denaturing agent was rapidly removed to keep the protein in an unprocessed form, followed by later addition of Ca(2+) and Zn(2+) ions. This allowed controlling the enzyme activation, when it is needed. PMID- 12651105 TI - Purification and characterization of a recombinant pea cytoplasmic fructose-1,6 bisphosphatase. AB - Full-length cDNA encoding pea cytoplasmic fructose-1,6-bisphosphatase (cyFBPase) was cloned from a pea cDNA library. The cloned cDNA was introduced into the Escherichia coli expression vector pET-15b. The recombinant cyFBPase was expressed in E. coli BL21 (DE3) cells in a soluble form and purified to homogeneity by Ni(+)-NTA affinity chromatography. The identity of the recombinant cyFBPase was confirmed by SDS-PAGE and immunoblot analysis using a polyclonal anti-His tag antibody. The recombinant cyFBPase was active at neutral pH ranges (6.6-9.0) and thermostable as other cyFBPases. The activation energy (E(a)) and Arrhenius frequency factor were 17.4 kcal/mol and 2.6 x 10(12)/s, respectively. The K(M) and V(max) values of the recombinant enzyme were calculated as 10.47 microM and 109 micromol/min, respectively. In case of removal of histidine tag, the K(M) value was calculated as 5.03 microM. The recombinant enzyme was non competitively and competitively inhibited by AMP and fructose-2,6-bisphosphate, respectively. PMID- 12651106 TI - Overexpression and purification of recombinant atrial natriuretic peptide using hybrid fusion protein REF-ANP in Escherichia coli. AB - Atrial natriuretic peptide (ANP), a small peptide consisting of 28 amino acids, has been applied in clinical treatment for heart failure, but it can encounter proteolytic degradation during its expression in host cells. Therefore, it is usually reported that ANP was expressed as a part of fusion protein. The aim of our study was to use an overexpression system to express the fusion protein REF ANP and to optimize a purification method. First, Escherichia coli DH5alpha was transformed with constructed expression vector containing two tandem copies of ref-anp gene and the fusion protein REF-ANP was overexpressed in shaking flask culture. Subsequently, the inclusion bodies were purified with reverse phase chromatography and pooled fractions were lyophilized. After this step, REF-ANP can be solubilized under native conditions without urea. After cleavage reaction, the sample was subjected to size exclusion chromatography and then rANP was polished with reverse phase chromatography. The final purity of rANP was more than 98% and the recovery of rANP per liter of shaking flask culture was more than 3mg. Such methods as mass spectrometry, capillary isoelectrofocusing analysis, and N-terminal amino acid sequence were used to identify rANP. The capillary isoelectrofocusing analysis showed that the pI of ANP was about pH 9.7. In this study, an efficient refolding and purification process should make scaling-up procedures easier and more successful than earlier reports. Moreover, it is possible that the refolding and purification method along with the overexpression system described in this article may offer new ideas on optimizing expression and purification of other kinds of short peptides. PMID- 12651107 TI - Expression and purification of human ribosomal proteins S3, S5, S10, S19, and S26. AB - The cDNAs for the human ribosomal proteins S3, S5, S10, S19, and S26 were introduced into a pET-15b vector and recombinant proteins containing an N (His)(6)-fusion tag were expressed in high yields. To resolve the problem of frameshift during expression of S26 caused by the presence of tandem arginine codons in its mRNA that are rare in Escherichia coli, we substituted the rare AGA codon with the more frequent arginine codon (CGC) using a primer with this mutation for PCR amplification of S26 cDNA. All proteins were expressed mainly in the form of inclusion bodies and purified to homogeneity by metal affinity chromatography in one step (except for S3). Expression of the full-length S3 was accompanied by the formation of a low molecular weight polypeptide that was co purified with S3 by metal affinity chromatography. Complete purification of S3 required an additional gel-filtration step. The proteins were refolded by stepwise dialysis. Both identity and purity of the proteins were confirmed by 2D PAGE. The proteins obtained could be used in a wide range of applications in biophysics, biochemistry, and molecular biology. PMID- 12651108 TI - Secretory expression in Escherichia coli and single-step purification of a heat stable alkaline protease. AB - Bacteriocin release proteins (BRPs) can be used for the release of heterologous proteins from the Escherichia coli cytoplasm into the culture medium. The gene for a highly thermostable alkaline protease was cloned from Bacillus stearothermophilus F1 by the polymerase chain reaction. The recombinant F1 protease was efficiently excreted into the culture medium using E. coli XL1-Blue harboring two vectors: pTrcHis bearing the protease gene and pJL3 containing the BRPs. Both vectors contain the E. coli lac promoter-operator system. In the presence of 40 microM IPTG, the recombinant F1 protease and the BRP were expressed and mature F1 protease was released into the culture medium. This opens the way for the large-scale production of this protease in E. coli. The recombinant enzyme was purified through a one-step heat treatment at 70 degrees C for 3h and this method purified the protease to near homogeneity. The purified enzyme showed a pH optimum of 9.0, temperature optimum of 80 degrees C, and was stable at 70 degrees C for 24h in the pH range from 8.0 to 10.0. The enzyme exhibited a high degree of thermostability with a half-life of 4 h at 85 degrees C, 25 min at 90 degrees C, and was inhibited by the serine protease inhibitor phenylmethylsulfonyl fluoride (PMSF). PMID- 12651109 TI - Construction and purification of his6-Thermus thermophilus MutS protein. AB - The mutS gene from the thermophilic bacterium Thermus thermophilus was PCR amplified, cloned, and expressed in Escherichia coli. The recombinant MutS protein containing an oligohistidine domain at the N-terminus was purified in a single step by Ni(2+) affinity chromatography to apparent homogeneity. The mismatch recognition properties of the his(6)-tagged MutS protein were confirmed by DNA protection against exonuclease digestion and retardation assays. The results of analytical gel filtration indicate that the predominant form of T. thermophilus MutS at micromolar concentrations is a tetramer. PMID- 12651110 TI - High-level expression of the Listeria monocytogenes listeriolysin O in Escherichia coli and preliminary characterization of the purified protein. AB - Listeriolysin O (LLO) is a cholesterol-binding sulfhydryl-activated hemolysin encoded by Listeria monocytogenes hlyA gene. After analyzing the nucleotide coding sequence of this gene from the ATCC 9525 L. monocytogenes strain, we cloned it in a pET vector for expression in Escherichia coli. Thanks to the optimization of the induction protocol, we achieved a high-level LLO synthesis (about 10% of total cell proteins) in hemolytically active form. The expressed hemolysin was then purified to homogeneity, as revealed by SDS-PAGE and Western blot analysis, by a hydroxyapatite adsorption chromatography, followed by an SP Sepharose ion-exchange chromatography. The recombinant protein showed the same properties determined for LLO purified from L. monocytogenes cultures and the characteristics of the sulfhydryl-activated toxins such as inactivation by oxidation and by reaction with cholesterol. By a combination of the pET expression system and the simple purification method, we obtained a significant amount of toxin (4.5 mg/litre cell culture) in a hemolytically active form (1.25 x 10(6)HU/mg protein). This procedure can solve the problem of LLO isolation from L. monocytogenes cultures, which is a difficult task, mainly owing to the low levels of toxin released in the culture media. The recombinant hemolysin, purified in sufficient quantities, could be very useful for structural studies and for diagnostic and pharmaceutical applications. PMID- 12651111 TI - High-level synthesis of Johnson grass mosaic virus coat protein in Escherichia coli and its auto-assembly to form virus-like particles. AB - The coat protein (CP) of Johnson grass mosaic virus (JGMV) auto-assembles to form virus-like particles (VLPs) and hence could be useful for presenting small peptides to the immune system. We are therefore attempting to synthesize JGMV CP in large amounts in Escherichia coli. The JGMV CP-encoding DNA, cloned under the bacteriophage T7 promoter, showed only low levels of CP synthesis in E. coli. The predicted secondary structure of the CP mRNA showed that its translational initiation codon was part of a stable hairpin-loop structure. The initiation codon could be relieved of the hairpin-loop structure by substitution of three neighboring nucleotides. This resulted in a single amino acid change at the N terminus of the protein. The modified RNA translated very efficiently, resulting in at least 16-fold higher CP accumulation in E. coli. The N-terminal amino acid substitution did not affect CP folding, as it auto-assembled in E. coli to form VLPs. PMID- 12651112 TI - Cloning, expression, purification, and activity of dog (Canis familiaris) and monkey (Saimiri boliviensis) cathepsin S. AB - Cathepsin S is the key protease responsible for the removal of the invariant chain from MHC class II molecules and, as such, has attracted much attention as a target for developing immunosuppressive drugs. To help in testing candidate compounds, the monkey (Saimiri boliviensis) and dog (Canis familiaris) cathepsin S cDNAs have been cloned. The monkey cDNA sequence encodes a 330 amino acid protein with 95% homology to human cathepsin S. The dog cDNA sequence encodes a 331 amino acid protein with 91% homology to human cathepsin S. The amino acid differences do not have a major effect on the hydrolysis of the substrate Z-VVR AMC, but may affect the substrate specificity. As for human and bovine cathepsin S, activity against Z-VVR-AMC extends into the neutral pH range. These parameters are important for understanding the role of cathepsin S in different species and for testing inhibitors in animal models of autoimmunity. PMID- 12651113 TI - High-level expression of a lipase from Bacillus thermocatenulatus BTL2 in Pichia pastoris and some properties of the recombinant lipase. AB - The BTL2 lipase gene from Bacillus thermocatenulatus was subcloned into the pPICZalphaA vector and integrated further into the genome of Pichia pastoris GS115. One of the best transformants harboring the linearized plasmid pPalpha BTL2 integrating into the P. pastoris genomic DNA was cultivated in a 5-L bioreactor filled with 4L of the culture medium BMMY. The BTL2 lipase was produced as an extracellular protein in large quantities of 309,000U/L supernatant. The lipase was purified using butyl-Sepharose with a specific activity of 23,000U/mg protein towards tributyrin. The pure enzyme was characterized and its physicochemical properties were compared to those of the BTL2 lipase, which had previously been expressed in Escherichia coli under the control of its native promoter on pUC18 or under the control of the strong temperature inducible promoter lambdaP(L), yielding 600U/g or 54,000U/g wet cells, respectively. The three proteins showed the same N-terminal sequence and had very similar pH optimum, pH stability, temperature optimum, thermostability, and substrate specificity profiles. Three enzymes were extremely stable in the presence of several organic solvents and detergents. PMID- 12651114 TI - Pyroglutamyl-peptidase I: cloning, sequencing, and characterisation of the recombinant human enzyme. AB - Pyroglutamyl-peptidase I (EC 3.4.19.3) is well known from bacteria and archaea, but has not previously been cloned or sequenced from any vertebrate. We describe the cloning and sequencing of the human (AJ278828) and mouse (AJ278829) forms of pyroglutamyl-peptidase I. The deduced amino acid sequences each consist of 209 residues and show approximately 30% identity with bacterial forms of the enzyme. They show clear homology to the enzyme from prokaryotes and place the mammalian forms of the enzyme in peptidase family C15 of the MEROPS database. The catalytic residues Glu81, Cys144, and His166 in the enzyme from Bacillus amyloliquefaciens are all conserved in the human sequence. A simple cartoon model of the human protein was constructed on the basis of the published crystal structures of pyroglutamyl-peptidase I forms from Thermococcus litoralis and B. amyloliquefaciens. The human enzyme was expressed by use of a baculovirus vector in Spodoptera frugiperda cells. The recombinant protein was enzymatically active and had properties similar to those described for the naturally occurring mammalian enzyme. Gel-filtration chromatography of the active enzyme gave a molecular mass of about 24kDa, showing that the enzyme is active as the monomer. This contrasted with indications that the prokaryotic enzymes may be tetrameric. Recombinant human pyroglutamyl-peptidase I was active on pGlu-aminomethylcoumarin in the range pH 6-9, with maximal activity being seen at pH 7.0-8.5; it showed an absolute requirement for a thiol-reducing agent. In crude preparations, the enzyme was completely stable for 90 min at 50 degrees C. The enzyme was inhibited by transition metal ions including Ni(2+), Zn(2+), and Cu(2+), and by sulfhydryl blocking agents. Reversible inhibition was seen with 2-pyrrolidone (K(i)=50 microM), and surprisingly, with N-ethylmaleimide (K(i)=30 microM). PMID- 12651115 TI - A poly-His tag method for obtaining the C-terminal lobe of human transferrin. AB - Human serum transferrin is an essential bilobal protein that transports iron in the circulation for delivery to iron-requiring cells. Obtaining the C-terminal lobe of human transferrin in verified native conformation has been problematic, possibly because its 11 disulfide bonds lead to misfolding when the lobe is expressed without its accompanying N-lobe. A recently reported method for preparing the C-lobe free of extraneous residues, with normal iron-binding properties and capable of delivering iron to cells, makes use of a Factor Xa cleavage site inserted into the interlobal connecting strand of the full-length protein. An inefficient step in this method requires the use of ConA chromatography to separate the cleaved lobes from each other, since only the C lobe is glycosylated. Inserting a 6-His sequence near the start of the N-lobe enhances recovery of the recombinant transferrin from other proteins in the culture medium of the BHK21 cells expressing the mutant transferrin. The new procedure is more economical in time and effort than its predecessor, and offers the additional advantage of isolating C-lobe expressed with or without its glycan chains. PMID- 12651116 TI - Functional reverse transcriptase encoded by the human LINE-1 from baculovirus infected insect cells. AB - The human LINE-1 ORF2, which encodes reverse transcriptase, was inserted into a baculovirus shuttle vector and expressed in Sf 21 cells. An immunoreactive polypeptide (149kDa) synthesized by infected cells had reverse transcriptase activity. A procedure for purification of functional ORF2 protein from insect cells was developed. The enzyme was purified with good recovery to near homogeneity and retained stable DNA polymerase activity. The optimum reaction conditions of the enzyme were determined with respect to salts, pH, and temperature. Substrate specificities and divalent cation requirements were investigated. The recombinant enzyme had a 3-fold preference for Mg2+ over Mn2+ for reverse transcriptase activity on poly(rA).oligo(dT)(12). As for DNA synthesis, the recombinant ORF2 protein was found to possess both RNA-dependent and DNA-dependent DNA polymerase activities. PMID- 12651117 TI - High-level expression of a novel FMN-dependent heme-containing lyase, phenylacetaldoxime dehydratase of Bacillus sp. strain OxB-1, in heterologous hosts. AB - We examined the overexpression of a novel FMN-dependent heme-containing lyase, phenylacetaldoxime dehydratase (Oxd) of Bacillus sp. strain OxB-1, in Escherichia coli and Bacillus subtilis. Several plasmids were constructed to express the enzyme under the control of the lac promoter or its own promoter, together with or without nitrilase and a possible regulatory protein that is present in the wild-type genome. The enzyme was expressed using E. coli transfected with the plasmid pOxD-9OF. Expression was under the control of the lac promoter in the pUC18 vector and was much more effective when the start codon was changed from TTG to ATG. When the transfected cells were grown at 37 degrees C, the enzyme was produced mainly in inactive inclusion bodies, whereas the enzyme was largely soluble and active when the cells were grown at 30 degrees C. The production of active enzyme was markedly enhanced by increasing the volume of culture medium. This had the effect of slowing the rate of apoenzyme synthesis. A slow rate of synthesis allows for a more efficient incorporation of heme cofactor into the apoenzyme than a fast rate of synthesis. Under optimized conditions, the enzyme was produced in an active and soluble form at 15,000U/L of culture, which is about 1500-fold higher than the amount produced by the wild-type strain. Moreover, the enzyme comprised over 40% of total extractable cellular protein. PMID- 12651118 TI - Expression and purification of the dihydrolipoamide acetyltransferase and dihydrolipoamide dehydrogenase subunits of the Escherichia coli pyruvate dehydrogenase multienzyme complex: a mass spectrometric assay for reductive acetylation of dihydrolipoamide acetyltransferase. AB - Plasmids were constructed for overexpression of the Escherichia coli dihydrolipoamide acetyltransferase (1-lip E2, with a single hybrid lipoyl domain per subunit) and dihydrolipoamide dehydrogenase (E3). A purification protocol is presented that yields homogeneous recombinant 1-lip E2 and E3 proteins. The hybrid lipoyl domain was also expressed independently. Masses of 45,953+/-73Da (1 lip E2), 50,528+/-5.5Da (apo-E3), 51,266+/-48Da (E3 including FAD), and 8982+/ 4.0 (lipoyl domain) were determined by MALDI-TOF mass spectrometry. The purified 1-lip E2 and E3 proteins were functionally active according to the overall PDHc activity measurement. The lipoyl domain was fully acetylated after just 30 s of incubation with E1 and pyruvate. The mass of the acetylated lipoyl domain is 9019+/-2Da according to MALDI-TOF mass spectrometry. Treatment of the 1-lip E2 subunit with trypsin resulted in the appearance of the lipoyl domain with a mass of 10,112+/-3Da. When preincubated with E1 and pyruvate, this tryptic fragment was acetylated according to the mass increase. MALDI-TOF mass spectrometry was thus demonstrated to be a fast and precise method for studying the reductive acetylation of the recombinant 1-lip E2 subunit by E1 and pyruvate. PMID- 12651119 TI - An unusual halotolerant alpha-type carbonic anhydrase from the alga Dunaliella salina functionally expressed in Escherichia coli. AB - A 60-kDa, salt-inducible, internally duplicated alpha-type carbonic anhydrase (Dca) is associated with the plasma membrane of the extremely salt-tolerant, unicellular, green alga Dunaliella salina. Unlike other carbonic anhydrases, Dca remains active over a very broad range of salinities (0-4M NaCl), thus representing a novel type of extremely halotolerant enzyme. To elucidate the structural principles of halotolerance, structure-function investigations of Dca have been initiated. Such studies require considerable amounts of the enzyme, and hence, large-scale algal cultivation. Furthermore, the purified enzyme is often contaminated with other, co-purifying algal carbonic anhydrases. Expression in heterologous systems offers a means to produce, and subsequently purify, sufficiently large amounts of Dca required for activity and structural studies. Attempts to over-express Dca in the Escherichia coli BL21(DE3)pLysS strain, after optimizing various expression parameters, produced soluble, but weakly active protein, composed of fully reduced and variably -S-S- cross-linked chains (each of the Dca repeats contains a pair of cysteine residues, presumably forming a disulfide bond). However, when the E. coli Origami B(DE3)pLysS strain was used as a host, a functionally active enzyme with proper disulfide bonds was formed in good yield. Affinity-purified recombinant Dca resembled the native enzyme from D. salina in activity and salt tolerance. Hence, this expression system offers a means of pursuing detailed studies of this extraordinary protein using biochemical, biophysical, and crystallographic approaches. PMID- 12651120 TI - Rapid purification of recombinant betaB2-crystallin using hydrophobic interaction chromatography. AB - BetaB2-crystallin, the major subunit of beta-crystallins, is difficult to purify either from lens homogenate or from betaH-or betaL-crystallins. It has been prepared by heterologous expression in Escherichia coli. Most often, the methods used for purifying a recombinant globular protein employ the combination of ion exchange with gel filtration chromatography. In the case of betaB2-crystallin too, different approaches have been used to obtain the purified protein, majority of which use a combination of ion-exchange and gel filtration chromatography. We present a new approach to purify betaB2-crystallin using hydrophobic interaction chromatography. In this method, the protein is bound to the hydrophobic matrix in the presence of high concentration of a non-chaotropic salt and eluted by decreasing the salt concentration. The method that we have used for the purification of this globular protein has definite advantages over the earlier methods in its simplicity and efficiency. The most noted advantage of this procedure is the rapid purification with a relatively purified product and a comparatively high yield (>20 mg/L of culture). Over all, the present protocol provides a rapid, efficient and simplified procedure for the preparation of betaB2-crystallin in large yield, sufficient for structural and functional studies. PMID- 12651121 TI - HIV-1 protease variants from 100-fold drug resistant clinical isolates: expression, purification, and crystallization. AB - High-resolution X-ray crystallographic structures of HIV-1 protease clinical variants complexed with licensed inhibitors are essential to understanding the fundamental cause of protease drug resistance. There is a need for structures of naturally evolved HIV-1 proteases from patients failing antiretroviral therapy. Here, we report the expression, purification, and crystallization of clinical isolates of HIV-1 protease that have been characterized to be more than 100 times less susceptible to US FDA approved protease inhibitors. PMID- 12651122 TI - Expression of proteins using the third domain of the Escherichia coli periplasmic protein TolA as a fusion partner. AB - The third domain of the periplasmic protein TolA from Escherichia coli (TolAIII) was used as a fusion partner in the expression of various proteins from bacteria and eukaryotes. TolAIII is small domain, expressed in high yields as a soluble protein in the cytoplasm of E. coli. Proteins were linked to the C-terminus of TolAIII by a short flexible linker containing sites for endopeptidases. Three different vectors were prepared, containing sites for enterokinase, thrombin or factor Xa. Fusion proteins also contain a His(6)-Ser(2) tag at their N-terminus for easier purification. Up to 90 mg fusion protein per liter bacterial culture was obtained using these vectors. Colicin N R-domain was expressed with this system as a fusion and processed further for functional studies. The yield of final pure R-domain was doubled as compared to the direct expression. The system may prove to be useful in the preparation of other peptides and proteins. PMID- 12651123 TI - Purification and properties of 2-hydroxy-6-oxo-6-(2'-aminophenyl)hexa-2,4-dienoic acid hydrolase involved in microbial degradation of carbazole. AB - Hydrolysis following meta-ring cleavage by a dioxygenase is a well-known step in aromatic compound metabolism. The 2-hydroxy-6-oxo-6-(2'-aminophenyl)hexa-2,4 dienoic acid hydrolase from Pseudomonas LD2 is a new member of the small group of characterized aromatic hydrolases that catalyze the cleavage of C-C bonds. In this study, the His(6)-tagged 2-hydroxy-6-oxo-6-(2'-aminophenyl)hexa-2,4-dienoic acid (HOPDA) hydrolase was purified from a recombinant Escherichia coli strain utilizing immobilized metal affinity chromatography. 2-Hydroxy-6-oxo-6-(2' aminophenyl)hexa-2,4-dienoic acid hydrolase is a colorless homodimer with no cofactor requirement. The enzyme actively converted HOPDA into benzoic acid and 2 hydroxypenta-2,4-dienoic acid. The enzyme exhibited activity between pH 6.5 and 10.5 with a maximum activity at pH 7.0. The optimum temperature at pH 7.0 was 60 degrees C. The calculated K'(m) for HOPDA was 4.6 microM, the V(max) was 3.3 micromol min(-1), and the K(s) was 70.0 microM. This corresponds to a maximum specific turnover rate of 1300 HOPDAs(-1)dimer(-1). The deduced amino acid sequence of CarC showed 30.3, 31.3, and 31.8% identity with TodF (P. putida F1), XylF (P. putida), and DmpD (Pseudomonas sp. CF600), respectively, which are meta cleavage compound hydrolases from other Pseudomonads. The amino acid sequence Gly X-Ser-X-Gly, which is highly conserved in these hydrolases, is also found in CarC. Lysates from a strain expressing enzyme in which the putative active site serine is mutated to alanine showed a significant reduction in activity. PMID- 12651125 TI - Separation between the alpha and beta forms of human antithrombin by hydroxyapatite high-performance liquid chromatography. AB - Human antithrombin (AT) inhibits several proteases in the coagulation system, including thrombin and factor Xa, and thus, plays an important role in the regulation of blood coagulation. The predominant form of AT in plasma is ATalpha, which contains four glycosylated asparagine residues, and the minor form is ATbeta, which lacks the Asn-135 glycosylation. In this study, hydroxyapatite high performance liquid chromatography, using a segmented sodium phosphate gradient, was utilized for the high-resolution separation of ATalpha and ATbeta. The detection limit (signal-to-noise ratio of 3) for ATbeta was 30 microg/mL, corresponding to 0.5% of the injected concentration of AT. Two analyzed commercial AT products both contained about 2% ATbeta. This method is suitable for the determination of ATbeta in pure samples of native AT. PMID- 12651124 TI - Macroaffinity ligand-facilitated three-phase partitioning for purification of glucoamylase and pullulanase using alginate. AB - Starch-degrading enzymes glucoamylase (from Aspergillus niger), and pullulanase (from Bacillus acidopullulyticus) were purified using alginates (polysaccharides consisting of mannuronic acids and guluronic acids) by a recently developed technique called macroaffinity ligand-facilitated three-phase partitioning (MLFTPP). In this process, a crude preparation of the enzyme was mixed with alginate. On addition of appropriate amounts of ammonium sulfate and t-butanol, the alginate bound enzyme appeared as an interfacial precipitate between the lower aqueous and the upper t-butanol phase. Enzyme activity from this interfacial precipitate was recovered using 1M maltose. Glucoamylase and pullulanase were purified 20- and 38-fold with 83% and 89% activity recovery, respectively. Both the purified preparations showed a single band on SDS-PAGE. PMID- 12651126 TI - Fast purification of the Apo form and of a non-binding heme mutant of recombinant sperm whale myoglobin. AB - As molecular biology has developed it has become possible to abundantly produce heterologous proteins in bacteria and to design serial amino acid substitutions for the generation of modified proteins, an approach also known as protein engineering. Sperm whale myoglobin, a protein of broad interest, has been cloned for several years now and a large collection of mutants has been produced. The presence of heme stabilizes the protein, which is recovered soluble from the bacterial pellet, and most purification protocols take advantage of this property for myoglobin purification directly from the pellet. However, recovery from the column resin is poor with these methods making them expensive and the procedure for removing heme is laborious and drastic when the apo form of Mb is required. In the case of proteins with severe mutations, which bind heme weakly or do not bind it at all, such methods cannot be employed without massive loss of productivity. Here, we describe a modified method, which is both low cost and rapid, for the purification of the soluble apo form of Mb from Escherichia coli inclusion bodies. Biophysical characterization of the protein after purification shows that the purified apoMb retains its native conformation and is soluble. This modified method is also used for the purification of a non-heme-binding apoMb mutant, demonstrating its efficiency when dealing with drastic mutations. PMID- 12651128 TI - Carbon-mineral adsorbents from waste materials: case study. AB - Waste bleaching earth from the food industry obtained in the process of fruit juice purification was utilized for preparation of carbon-mineral adsorbents. The waste material, containing 25.8 wt% C, was subjected to three kinds of treatment: (1) direct pyrolysis at 400 degrees C with a suitable temperature program; (2) preliminary hydrothermal modification (200 degrees C, 8 h, 15.3 atm) and then pyrolysis as in method 1; (3) preliminary thermal treatment (400 degrees C) and then chemical treatment (boiling in 3% solution Na(2)CO(3)), followed by heating at 400 degrees C (10 min). Moreover, the materials obtained by these methods were subjected to additional thermal treatment at 700 degrees C with a suitable temperature program. Both the morphology and the topography of carbon deposits and, in consequence, the porous structure of the obtained adsorbents depend on the method of their preparation. The additional thermal treatment of these samples at 700 degrees C makes it possible to obtain adsorbents of more thermally stable carbon deposits possessing better parameters of the porous structure. Carbon-mineral adsorbents of different specific surface areas (S(BET) from 17.6 to 153 m(2)/g) and pore volumes (from 0.035 to 0.093 cm(3)/g) were prepared. The mechanism of phenol and p-nitrophenol adsorption on the obtained adsorbents was discussed and their properties were compared with the suitable literature data. PMID- 12651130 TI - Alpha-lactalbumin tertiary structure changes on hydrophobic interaction chromatography surfaces. AB - Hydrogen exchange (HX) detected by mass spectrometry (MS) was used to analyze the structure of calcium-free alpha-lactalbumin, a model protein with marginal stability. Two chromatographic peaks were observed from samples of pure protein eluted from SOURCE phenyl hydrophobic interaction chromatography (HIC) media. Whole-protein HX measurements showed that the less-retained peak had solvent exposure similar to native protein in the absence of the HIC surface while the retained protein was nearly, although not fully, solvent exposed. The formation of these two peaks was kinetically limited. The protein also refolded successfully following elution. In addition, proteolytic fragmentation was used to analyze HX at the peptide level. This approach revealed that helix C was the most stable region of alpha-lactalbumin under native conditions and in the flow through peak. Helix C also formed the core of residual native structure in the partially unfolded protein in the retained peak. The results suggest that residues that are most solvent accessible under native conditions may be those most likely to unfold upon adsorption. PMID- 12651129 TI - Immobilization and surface characterization of NeutrAvidin biotin-binding protein on different hydrogel interlayers. AB - For a number of potential applications, it is desirable to immobilize avidin class molecules onto solid supports and exploit their ability to bind biotinylated molecules with high affinity. NeutrAvidin molecules were surface immobilized in various ways. In this study, NeutrAvidin was covalently attached by carbodiimide chemistry onto carboxyl groups of polyacrylic acid and carboxymethyl-dextran hydrogel interlayers. A third strategy involved the affinity "docking" of NeutrAvidin onto a biotinylated poly(ethylene glycol) interlayer. These three interlayers were selected for their low nonspecific binding of proteins, which was expected to minimize surface binding of NeutrAvidin by nonspecific interfacial adsorption. X-ray photoelectron spectroscopy (XPS) analyses allowed detailed characterization of the multilayer fabrication steps. An ELISA assay was used to measure NeutrAvidin activity, which varied with the surface immobilization route. Atomic force microcopy (AFM) force measurements showed that the hydrogel interlayer contributed to a repulsive force and verified the specific interaction between biotinylated AFM tips and the NeutrAvidin surfaces. When a solution of free biotin was injected into the AFM liquid cell, the force curve changed substantially and became identical to that recorded between surfaces carrying no NeutrAvidin, indicating that the free solution biotin had displaced NeutrAvidin proteins off the PEG-biotin layer. PMID- 12651131 TI - Aggregation of sphalerite: role of zinc ions. AB - Sphalerite from six sources is shown to aggregate at ca. pH 7-9, confirmed by different techniques (settling velocity, suspension analysis, and optical microscopy). This does not correlate with the isoelectric point, which is consistently < pH 6. A similar observation was made by Healy and Jellet [J. Colloid Interface Sci. 24 (1967) 41-46] for ZnO (and reconfirmed here), who suggested Zn hydrolysis products were responsible. The hypothesis is tested using chalcopyrite and silica suspensions in the presence of Zn ions. It is found that aggregation occurs over the same pH range, 7-9. Solution and surface analysis (extraction by EDTA) shows sphalerite released sufficient Zn ions to promote aggregation. The pH range 7-9 corresponds to hydroxide formation, suggesting this is the species responsible. Aggregation of sphalerite due to the polymerization/flocculation action of zinc hydroxide is proposed. PMID- 12651132 TI - Synthesis of uniform anatase TiO2 nanoparticles by gel-sol method. 3. Formation process and size control. AB - Uniform anatase-type TiO(2) nanoparticles were prepared by the gel-sol process from a condensed Ti(OH)(4) gel preformed by the hydrolysis of a Ti triethanolamine (TEOA) complex. The particle size of the anatase TiO(2) was increased from ca. 5 to 30 nm with pH increasing from 0.6 to 12 by aging the Ti(OH)(4) gel at 140 degrees C for 72 h, while the yield of the anatase TiO(2), 100% below pH 9.6, started to decrease from pH 10, to 67% at pH 11.5 and only 9% at pH 12.2. These results reveal a significant reduction of the nucleation rate of the anatase TiO(2) with increasing pH, as is explained by the reduction of the concentration of a precursor complex, Ti(OH)(3)(+), and the adsorption of hydroxide ion onto the embryos of TiO(2). Triethanolamine appeared to enhance the pH effect on the nucleation rate of anatase TiO(2) particles by adsorption onto their embryos, leading to the wide range of the size control. Triethanolamine was also found to act as a shape controller of the anatase TiO(2) particles for yielding ellipsoidal particles from Ti(OH)(4) gel at a relatively high pH above 11. The particle size was also controlled by seeding of anatase TiO(2). Moreover, the seeding experiment suggested that the rate-determining step of the gel-sol process was not the dissolution of the hydroxide gel, but the deposition of the monomeric precursor from the solution phase. PMID- 12651133 TI - Synthesis of uniform anatase TiO2 nanoparticles by gel-sol method. 4. Shape control. AB - Uniform anatase-type TiO(2) nanoparticles of different shapes have been formed by phase transformation of a Ti(OH)(4) gel matrix in the presence of shape controllers. For example, triethanolamine (TEOA) was found to change the morphology of TiO(2) particles from cuboidal to ellipsoidal at pH above 11. The shape control can be explained in terms of the specific adsorption of TEOA onto the crystal planes parallel to the c-axis of the tetragonal system in the alkaline range, as supported by the observation of preferential adsorption of TEOA onto the crystal planes parallel to the c-axis at pH 11.5 and by the pH dependence of the adsorption onto ellipsoidal particles. Diethylenetriamine (DETA) also modified the particle shape to ellipsoidal above pH 9.5 and the aspect ratio was much higher than with TEOA. The mechanism of the shape control could be explained in the same way as with TEOA, since analogous specific adsorption was observed with DETA as well. Similar shape control to yield ellipsoidal particles of a high aspect ratio was also achieved with other primary amines, such as ethylenediamine (ED), trimethylenediamine (TMD), and triethylenetetramine (TETA). However, secondary amines, such as diethylamine, and tertiary amines, such as trimethylamine and triethylamine, acted as a complexing agent of Ti(IV) ions to promote the growth of ellipsoidal particles of a low aspect ratio, rather than a shape controller to produce ellipsoids of a high aspect ratio. Sodium oleate and sodium stearate were found to modify the particle shape from round-cornered cubes to sharp-edged cubes. The mechanism was explained in terms of the reduction of the specific surface energies of the [001] and [100] planes of the tetragonal crystal system by the preferential adsorption of oleate or stearate ion onto these planes, based on the adsorption experiment using ellipsoidal and cubic particles. PMID- 12651135 TI - Effect of surface mobility on the particle sliding along a bubble or a solid sphere. AB - The sliding velocity of glass beads on a spherical surface, made either of an air bubble or of a glass sphere held stationary, is measured to investigate the effect of surface mobility on the particle sliding velocity. The sliding process is recorded with a digital camera and analyzed frame by frame. The sliding glass bead was found to accelerate with increasing angular position on the collector's surface. It reaches a maximum velocity at an angular position of about 100 degrees and then, under certain conditions, the glass bead leaves the surface of the collector. The sliding velocity of the glass bead depends strongly on the surface mobility of a bubble, decreasing with decreasing surface mobility. By a mobile surface we mean one which cannot set up resistive forces to an applied stress on the surface. The sliding velocity on a rigid surface, such as a glass sphere, is much lower than that on a mobile bubble surface. The sliding velocity can be described through a modified Stokes equation. A numerical factor in the modified Stokes equation is determined by fitting the experimental data and is found to increase with decreasing surface mobility. Hydrophobic glass beads sliding on a hydrophobic glass sphere were found to stick at the point of impact without sliding if the initial angular position of the impact is less than some specific angle, which is defined as the critical sticking angle. The sticking of the glass beads can be attributed to the capillary contracting force created by the formation of a cavity due to spontaneous receding of the nonwetting liquid from the contact zone. The relationship between the critical sticking angle and the particle size is established based on the Yushchenko [J. Colloid Interface Sci. 96 (1983) 307] analysis. PMID- 12651134 TI - Investigation of a colloidal damper. AB - A novel application of nanotechnology in the field of mechanical engineering, called colloidal damper (CD), is investigated. This device is complementary to the hydraulic damper (HD), having a cylinder-piston construction. Particularly for CD, the hydraulic oil is replaced by a colloidal suspension, which consists of a mesoporous matrix and a lyophobic fluid. In this work, the porous matrix is from silica gel modified by linear chains of n-alkylchlorosilanes and water is considered as an associated working fluid. A design solution from a practical point of view of the CD test rig and the measuring technique of the hysteresis are described. A brief review of the water physical properties relative to the CD concept is presented. Influence of the bonding density, length of the grafted molecule, pore diameter, and particle diameter on the CD hysteresis is investigated for distinctive types and mixtures of silica gels. Temperature variation during functioning is recorded and the CD cycle is interpreted from a thermodynamic standpoint. Variation of the CD dissipated energy and efficiency with pressure, water quantity, and relaxation time is illustrated. Experimental results are justified by the analysis of the water flow into the porous matrix, CD thermodynamics, and the mechanism of the energy dissipation. Our findings agree with the previously published data. PMID- 12651136 TI - Reversible charging of the ice-water interface. II. Estimation of equilibrium parameters. AB - Equilibrium parameters characterizing the ice-water interface were obtained from electrokinetic and potentiometric data. The dependency of surface potential on pH was measured by the ice electrode. Electrokinetic zeta potentials were measured by electrophoresis. In the interpretation, the 1-pK and 2-pK concepts of the surface complexation model, developed for metal oxides, were used. The equilibrium parameters (constants of surface reactions) were determined. The results were confirmed by numerical simulations. Both 1-pK and 2-pK models fit the experimental data well. PMID- 12651137 TI - Pitting inhibition of stainless steel by surfactants: an electrochemical and surface chemical approach. AB - Pitting corrosion of stainless steels causes tremendous damage in terms of material loss and resulting accidents. Organic surfactants have been tried as pitting inhibitors but the understanding of the inhibition mechanisms is mainly speculative. In the present study the inhibition of the pitting corrosion of 304 stainless steel by N-lauroylsarcosine sodium salt (NLS) in 0.1 M NaCl solutions at neutral pH was studied using an approach that combines surface chemical techniques with electrochemical ones. It was found that NLS increases the pitting resistance of 304 stainless steel, with possible complete inhibition at high NLS concentration (30 mM). Adsorption of NLS on 304 stainless steel particles was directly measured. NLS adsorbs significantly on 304 stainless steel with maximum adsorption density close to bilayer coverage. Electrophoretic mobility data for 304 stainless steel particles show that the surface of 304 stainless steel is negative in NaCl solution at neutral pH. The adsorption of NLS makes the interfacial charge even more negative. The relationship between pitting inhibition and adsorption density of NLS suggests that NLS does not adsorb preferentially on the pit nucleation sites and complete inhibition requires that the whole surface be covered completely by NLS. The inhibition mechanism of NLS is proposed to be due mainly to the blocking effect of a negatively charged NLS adsorption layer. This study shows that in addition to the adsorption amount of surfactant, interfacial charge also plays an important role in pitting inhibition. PMID- 12651138 TI - Interfacial interactions between amphiphilic cyclodextrins and physiologically relevant cations. AB - The compression isotherms of a series of amphiphilic cyclodextrins, formed (a) by acylation at the secondary hydroxyl face and (b) by acylation accompanied by varying degrees of sulfatation (DS) at the primary hydroxyl face (DS=0, 4, and 7), have been studied on subphases of pure water and of water containing NaCl, KCl, MgCl(2), and CaCl(2) at inter- and extracellular concentrations. The formation of solid lipid nanoparticles (SLNs) by two of the molecules has been observed, while these do not aggregate at concentrations of monovalent salts up to 150 mM for the sulfated derivative. In the presence of divalent salts one of these with a DS=0 for sulfatation degree flocculates at divalent salt concentrations below 0.1 mM while the other with a DS=4 flocculates at Mg(2+) concentration above 5 mM and a Ca(2+) concentration above 3 mM. AFM noncontact mode imaging has been carried out, in air, for the SLNs deposited on mica. PMID- 12651139 TI - Early stages of particle formation in precipitation reactions-quinacridone and boehmite as generic examples. AB - For many products, such as nanoparticulate systems, particle formation by precipitation is an essential procedural step. To learn more about the processes involved in precipitation, we investigated particle formation during precipitation reactions by means of online and offline methods. As model systems we chose the catalyst boehmite and the organic pigment quinacridone. The reactants were mixed in a mixing device and led into a reaction tube. At the end of the tube, a free jet of the suspension was produced. By varying the length of the reaction tube the time between mixing the reactants and the moment of observation was varied. Thus a time resolution down to 10 ms from the beginning of the reaction was obtained. Small-angle X-ray scattering on the free jet yielded online information about the structural inhomogeneities within the reacting systems. Transmission electron microscopy patterns obtained from quenched samples, which were taken by shooting copper grids through the free jet into liquid nitrogen, provided complementary information about structural features. Immediately after mixture an emulsion-like structure develops indicating that classical nucleation theory does not apply in the present systems. This finding can be explained by assuming instantaneous reaction at the interfaces of the two reactants that meet in the mixing device. From this preliminary state primary particles form with a size in the nanometer range. The observations can be rationalized by considering the underlying hydrodynamics of turbulent mixing of the reactants. PMID- 12651140 TI - Crystallization behaviors of nanosized MgO particles from magnesium alkoxides. AB - The effects of the size of the alkoxy group on the thermal decomposition behavior of magnesium alkoxides (magnesium methoxide and ethoxide) and the crystallization behavior of MgO was investigated using thermogravimetry, Fourier-transformed infrared spectroscopy, X-ray powder diffraction, and transmission electron microscopy. As the size of the alkyl group increased, the decomposition temperature decreased and resultant MgO crystallization of the alkoxide precursor was enhanced. In an inert N(2) atmosphere, the decomposition temperature of magnesium ethoxide was about 260 degrees C, which was lower than that of magnesium methoxide by approximately 70 degrees C. The degree of the crystallization of MgO particles from the ethoxide was also significantly higher than that of the methoxide. This result is explained in terms of the O-R bonding strength of the alkoxide. With use of the Kissinger method, the activation energy for the thermal decomposition of magnesium alkoxide was found to be dependent on the size of the alkyl group. The activation energies were 161+/-23 and 130+/-24 kJ/mol for the magnesium methoxide and the magnesium ethoxide, respectively. PMID- 12651142 TI - Limiting conditions for applying the spherical section assumption in contact angle estimation. AB - The shape of liquid drops on solid surfaces deviates from the spherical as tension decreases and gravity effects start affecting the drop shape. This paper attempts to define this deviation and estimates the dimensionless Eotvos number limits above which the deviation becomes "significant." The use of these limiting values can facilitate estimation of contact angle in the following manner. It is well known that the equilibrium contact angle made by a liquid drop on a solid surface can be estimated from measurements of two drop parameters. These parameters can be any two chosen from the drop volume, height, and wetted radius. In case the effect of gravity on the drop shape is negligible, simple algebraic relations derived from the spherical section assumption exist, from which the contact angle can be estimated. In systems where the "spherical section" assumption is invalid, the Laplace equation for the drop shape has been solved numerically with any two of the above parameters as the constraints, to obtain the contact angle. In this paper, Eotvos numbers at which the deviation of the drop profile from the spherical is significant enough to result in contact angle deviation of 1 degrees are estimated. The limiting values of Eotvos number, expressed as a function of the original contact angle made by the spherical profile, are obtained by solving the Laplace equation for the drop shape with the drop volume and wetted radius constraints for decreasing values of Interfacial tension. These limiting values are also estimated for different drop sizes and for cases where the drop phase is heavier (sessile) and lighter (buoyant) than the surrounding fluid. The independence of the Eotvos number estimates from the sign of the density difference as well as the drop size is shown. These Eotvos number limits can be used to check if the spherical section assumption, with the resulting simple algebraic relations, can be used for contact angle estimation and other shape-related analysis for a system. PMID- 12651143 TI - Temperature effect on solubilization of n-alkylbenzenes into sodium cholate micelles. AB - The solubilization of n-alkylbenzenes (benzene, toluene, ethylbenzene, n propylbenzene, n-butylbenzene, n-pentylbenzene, n-hexylbenzene) into an aqueous micellar solution of sodium cholate was carried out. Solubilizate concentrations at equilibrium were determined spectrophotometrically at 293.2, 298.2, 303.2, 308.2, and 313.2 K. The first stepwise association constants (K(1)) between solubilizate monomers and vacant micelles were evaluated from the equilibrium concentrations and found to increase with increasing hydrophobicity of the solubilizate molecules. From the Gibbs energy change for solubilization at different micelle aggregation numbers and from the molecular structure of the solubilizates, the function of sodium cholate micelles as solubilizer was discussed. Enthalpy and entropy changes of solubilization were calculated from the temperature dependence of the K(1) values, and the solubilization was found to be enthalpy-driven for the solubilizates with shorter alkyl chains. The results obtained were also compared with those for conventional aliphatic micelles. PMID- 12651141 TI - Impregnated active carbons to control atmospheric emissions. I. Influence of the impregnated species on the porous structure. AB - Impregnated active carbons were prepared to be used as catalysts for complete oxidation, aiming at the reduction of atmospheric emission of volatile organic compounds. Good dispersion of the catalyst is required, as it regulates the conversion efficiency in the pores where pollutants can access to be converted. When impregnation is performed on the raw material or after activation, the influence of impregnated species on the structure and on the catalyst dispersion is already well studied. This paper aims to analyze the influence of impregnated species on the structure and on the catalyst dispersion when the impregnation step is performed after carbonization, as this knowledge is yet very scarce. Olive stones were used as raw material. In order to evaluate the influence of the methodology of impregnation with CoO, Co(3)O(4), and CrO(3), the impregnation step was completed after activation and between carbonization and activation. In the first sequence, the impregnated oxides must be deposited on the internal surface, blocking part of the initial microporous structure. When impregnation is conducted after carbonization, metal species act as catalysts during the activation step. The structure developed strongly depends either on the chemical state of the catalyst or on its distribution in the carbonized material. A well developed pore structure can appear just where metal species can access, remaining a micropore structure with pores of very small sizes. The better efficiency of Co(3)O(4) as a catalyst of the activation step is conditioned by a worse distribution of the carbonized material, related to the bigger size of its crystals. CrO(3) is the least efficient catalyst for the activation step. PMID- 12651144 TI - Temperature effect on formation of sodium cholate micelles. AB - The micellization of sodium cholate (NaC) at 293.2, 298.2, 303.2, 308.2, and 313.2 K by cholate anion concentration was studied over the pH range from 6.0 to 7.2. Using a stepwise association model of cholate anions without bound sodium counterions, the aggregation number (nmacr;) of the cholate micelles was evaluated and found to increase with the total concentration, indicating that the stepwise association model is applicable. The nmacr; values go up and down with increasing temperature; 17 at 298.2 and 12 at 313.2 K and at 60 mM of the sodium cholate. The fluorescence of pyrene was measured in sodium cholate solution to determine the critical micelle concentration (CMC), indicating a narrow concentration range for CMC. A sodium-ion-specific electrode was used to determine a relatively low degree of counterion binding to micelles, supporting the validity of the present association model of cholate anions. The aggregation numbers evaluated at a constant ionic strength of 0.15 and at lower but variable ionic strengths were similar except for higher cholate concentrations. PMID- 12651145 TI - Radiolabeling of Pluronic amphiphilic copolymer for adsorption studies. AB - A new pathway for the radiolabeling of Pluronic PE6800 was developed. In a first step, the CH(2)-OH end groups of the copolymer were substituted by tosylates; in a second step these were reduced by [3H]-NaBH(4) to obtain tritiated chain ends. The final product was shown to be a mixture of native, tosylated, and reduced Pluronic containing 1 tritium atom per 1110 Pluronic molecules. The labeling procedure did not affect the molecular weight distribution nor the adsorption isotherm of the copolymer on polystyrene plates. A plateau value of about 0.7 microg/cm(2) is reached at a concentration in solution of 500 microg/ml, i.e., much lower than the cmc. Upon drying, the Pluronic adsorbed layer reorganizes in particles with a size of about 30 to 60 nm which cover about 15% of the substratum surface. This observation is of great importance for the design of protein-resistant surfaces by adsorption of Pluronic. PMID- 12651146 TI - Correlation in the properties of aqueous single films and foam containing a nonionic surfactant and organic/inorganic electrolytes. AB - Correlation of the behavior of foam from aqueous solutions of C(10)E(8) + TPeAB mixtures with the properties of the single foam films is sought through a theoretical analysis based on experimental data. The state in the adsorption layer on the air/solution interface has been determined from the surface tension experimental data by applying the Frumkin adsorption isotherm for mixed surfactants. Diverse parameters of the adsorption layer have been calculated. Values of the surface potential, estimated from the electrostatic disjoining pressure in the films, are found to be in good agreement with those calculated through the Grahame equation for the air/water interface. The energy of interaction (attraction) in the mixed adsorption layers is determined. The behavior of a foam body produced from solutions of the same composition, is also investigated in two independent ways using the stationary foam column and the foam life-time (decay rate) methods. Both techniques gave similar results with respect to foam stability. PMID- 12651147 TI - Variation of bubble size distribution in a protein foam fractionation column measured using a capillary probe with photoelectric sensors. AB - Bubble size is used to characterize not only bubble-specific interfacial area but also bubble coalescence in a foam column. The bubble size distributions were obtained in a continuous foam fractionation process for concentrating ovalbumin using a developed photoelectric probe. When the continuous process reached steady state, the bubble size distribution pattern remained stable. Bubble size distribution data above (+1 cm) or below (-1 cm) the bulk liquid-foam interface showed symmetry along the diameter of the column (14 cm ID). The bubble size distribution was affected by the column wall. The nearly constant protein concentration distribution across the column cross-section indicated that the bubble flow distribution approached a flat profile across the column. A log normal bubble distribution pattern best fit the weighted range of bubbles in the column at column lengths above and below the liquid-foam interface. These observations may prove to be useful in understanding the mechanisms underlying the foam fractionation of proteins. PMID- 12651148 TI - Stability conditions of an electrified miscible viscous fluid sheet. AB - The Kelvin-Helmholtz problem of viscous fluids under the influence of a normal periodic electric field in the absence of surface charges is studied. The system is composed of a streaming dielectric fluid sheet of finite thickness embedded between two different streaming finite dielectric fluids. The interfaces permit mass and heat transfer. Because of the complexity of the considered system, a mathematical simplification is adopted. The weak viscous effects are taken into account so that their contributions are incorporated into the boundary conditions. Therefore, the equations of motion are solved in the absence of viscous effects. The boundary value problem leads to two simultaneous Mathieu equations of damped terms having complex coefficients. The symmetric and antisymmetric deformations reduced the coupled Mathieu equations to a single Mathieu equation. The classical stability criterion is found to be substantially modified due to the effect of mass and heat transfer. The analytical results are numerically confirmed. It is found that the sheet thickness and mass and heat transfer parameters have a dual influence on the stability criteria. It is also found that the field frequency has a stabilizing influence especially at small values of the wave number. In contrast to the case of a pure inviscid fluid, it is found that the uniform normal electric field plays a dual role in the stability criteria. This role depends on the choice of the numerical values of the physical parameters of the system under consideration. PMID- 12651149 TI - Viscosity function in polymer-modified asphalts. AB - Asphalt is a multidisperse micellar system with rheological behavior resembling that of a low-molecular-weight polymer. Nowadays, asphalt is frequently modified by blending it with various polymers. Such modified asphalt has rheological properties that differ from the properties of the base asphalt. It is quite common to study asphalt in dynamic experiments. Such studies, however useful, cannot reveal all characteristic features of polymer-modified asphalts. Asphalt modification by polymers is strongly manifested in the region of transitions from a viscoelastic fluid to the Newtonian fluid. The viscosity study in this region can reveal behavior characteristic of the used polymer modifier, thus complementing the dynamic studies of these materials. The viscosity of base asphalt modified by styrene-butadiene-styrene and by ethylene-vinyl acetate polymers (in several concentrations) is studied and discussed in this note. PMID- 12651150 TI - Commitment of 3T3-F442A cells to adipocyte differentiation takes place during the first 24-36 h after adipogenic stimulation: TNF-alpha inhibits commitment. AB - We studied the commitment of 3T3-F442A cells during stimulation with adipogenic serum or growth hormone. Confluent 3T3-F442A preadipocytes were incubated with adipogenic medium for increasing times; the number of adipose clusters, GPDH activity, and lipid accumulation were evaluated. Results show that cell commitment took place during the first 24-36 h after stimulation under adipogenic conditions. Then, cultures underwent a 2-fold increase in total cell number through selective multiplication of committed cells, followed by a dramatic decrease in colony-forming ability and 300- to 1000-fold raise in GPDH activity. Cell commitment was not modulated by insulin, but this hormone stimulated clonal expansion of committed cells and lipogenesis. Commitment was inhibited by TNF alpha at concentrations as low as 5 ng/ml, and by retinoic acid. The results show that TNF-alpha inhibits adipose conversion at two different levels: at concentrations as low as 5 ng/ml, it blocks commitment, and at concentrations of 100 ng/ml or higher the cytokine seems to block mitotic expansion and other steps of differentiation after cell commitment. The identification of a specific time for cell commitment would allow the study of the early genes that might regulate cell reprogramming into adipocytes. PMID- 12651151 TI - ER retention may play a role in sorting of the nuclear pore membrane protein POM121. AB - Integral membrane proteins of the nuclear envelope (NE) are synthesized on the rough endoplasmic reticulum (ER) and following free diffusion in the continuous ER/NE membrane system are targeted to their proper destinations due to interactions of specific domains with other components of the NE. By studying the intracellular distribution and dynamics of a deletion mutant of an integral membrane protein of the nuclear pores, POM121, which lacks the pore-targeting domain, we investigated if ER retention plays a role in sorting of integral membrane proteins to the nuclear envelope. A nascent membrane protein lacking sorting determinants is believed to diffuse laterally in the continuous ER/NE lipid bilayer and expected to follow vesicular traffic to the plasma membrane. The GFP-tagged deletion mutant, POM121(1-129)-GFP, specifically distributed within the ER membrane, but was completely absent from the Golgi compartment and the plasma membrane. Experiments using fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) demonstrated that despite having very high mobility within the whole ER network (D = 0.41 +/- 0.11 micro m(2)/s) POM121(1-129)-GFP was unable to exit the ER. It was also not detected in post-ER compartments of cells incubated at 15 degrees C. Taken together, these experiments show that amino acids 1-129 of POM121 are able to retain GFP in the ER membrane and suggest that this retention occurs by a direct mechanism rather than by a retrieval mechanism. Our data suggest that ER retention might be important for sorting of POM121 to the nuclear pores. PMID- 12651152 TI - Gap junction-mediated bystander effect in primary cultures of human malignant gliomas with recombinant expression of the HSVtk gene. AB - The ability of herpes simplex virus type 1 thymidine kinase (HSV-tk)-expressing cells incubated with ganciclovir (GCV) to induce cytotoxicity in neighboring HSV tk-negative (bystander) cells has been well documented. Although it has been suggested that this bystander cell killing occurs via the transfer of phosphorylated GCV, the mechanism(s) of this bystander effect and the importance of gap junctions for the effect of prodrug/suicide gene therapy in primary human glioblastoma cells remains elusive. Surgical biopsies of malignant gliomas were used to establish explant primary cultures. Proliferating tumor cells were characterized immunohistochemically and found to express glial tumor markers including nestin, vimentin, glial fibrillary acidic protein (GFAP), S-100, and gap junction protein connexin 43 (Cx43). Western blot analysis revealed the presence of phosphorylated isoforms of Cx43 and Calcein/DiI fluorescent dye transfer showed evidence of efficient gap junction communication (GJC). In order to study the effect(s) of prodrug/suicide gene therapy in these cultures, human glioblastoma cell cultures were transfected with the HSVtk gene for transient or stable expression. Ganciclovir treatment of these cultures led to >90% of cells dead within 1 week. Eradication of cells could be inhibited by the addition of alpha-glycyrrhetinic acid (AGA), a GJC inhibitor. In parallel experiments, AGA decreased the immunodetection of phosphorylated Cx43 as analyzed by Western blot and inhibited fluorescent dye transfer. In conclusion, these observations are consistent with GJC as the mediator of the bystander effect in primary cultures of human glioblastoma cells by the transfer of phosphorylated GCV from HSVtk gene transfected cells to untransfected ones. PMID- 12651153 TI - Loss and recovery of the blood-nerve barrier in the rat sciatic nerve after crush injury are associated with expression of intercellular junctional proteins. AB - The blood-nerve barrier in peripheral nerves is important for maintaining the environment for axons. Breakdown of the barrier by nerve injury causes various pathologies. We hypothesized that the breakdown and recovery of the blood-nerve barrier after injury are associated with the changes in the expression of intercellular junctional proteins. To test this hypothesis, we induced crush injuries in the rat sciatic nerve by ligation and analyzed spatiotemporal changes of claudin-1, claudin-5, occludin, VE-cadherin, and connexin43 by immunoconfocal microscopy and morphometry and compared them with changes in the permeability of the blood-nerve barrier by intravenous and local administration of Evans blue albumin (EBA). On day 1 after removal of the ligature EBA leaked into the connective tissue in the endoneurium and then the leakage gradually decreased and disappeared on day 7. On day 1 claudin-1, claudin-5, occludin, VE-cadherin, and connexin43 had totally disappeared from the perineurium and endoneurium. Thereafter, claudin-1, claudin-5, occludin, and VE-cadherin recovered from day 2, whereas connexin43 was redetected on day 5. These results indicate that the breakdown and following recovery of the blood-nerve barrier are closely associated with changes in the expression of claudins, occludin, VE-cadherin, and connexin43 and that the recovery time course is similar but nonidentical. PMID- 12651154 TI - Apoptosis-related fragmentation, translocation, and properties of human prothymosin alpha. AB - Human prothymosin alpha is a proliferation-related nuclear protein undergoing caspase-mediated fragmentation in apoptotic cells. We show here that caspase-3 is the principal executor of prothymosin alpha fragmentation in vivo. In apoptotic HeLa cells as well as in vitro, caspase-3 cleaves prothymosin alpha at one major carboxy terminal (DDVD(99)) and several suboptimal sites. Prothymosin alpha cleavage at two amino-terminal sites (AAVD(6) and NGRD(31)) contributes significantly to the final pattern of prothymosin alpha fragmentation in vitro and could be detected to occur in apoptotic cells. The major caspase cleavage at D(99) disrupts the nuclear localization signal of prothymosin alpha, which leads to a profound alteration in subcellular localization of the truncated protein. By using a set of anti-prothymosin alpha monoclonal antibodies, we were able to observe nuclear escape and cell surface exposure of endogenous prothymosin alpha in apoptotic, but not in normal, cells. We demonstrate also that ectopic production of human prothymosin alpha and its mutants with nuclear or nuclear cytoplasmic localization confers increased resistance of HeLa cells toward the tumor necrosis factor-induced apoptosis. PMID- 12651155 TI - PACE-1, a novel protein that interacts with the C-terminal domain of ezrin. AB - The ERM proteins (ezrin, radixin, moesin) together with merlin comprise a subgroup of the band 4.1 superfamily. These proteins act as membrane cytoskeletal linker proteins mediating interactions between the cytoplasmic domains of transmembrane proteins and actin. To better understand how the ERM proteins function to regulate these junctional complexes, a yeast 2-hybrid screen was undertaken using ezrin as a bait. We describe here the identification and cloning of a novel protein, PACE-1, which binds to the C-terminal domain of ezrin. Characterization of PACE-1 in human breast cancer cell lines demonstrates it to have two distinct intracellular localizations. A proportion of the protein is associated with the cytoplasmic face of the Golgi apparatus. This distribution is dependent upon the presence of the PACE-1 N-terminal myristoylation consensus sequence but is not dependent on an association with ezrin. In contrast, PACE-1 colocalises with ezrin in the lamellipodia, where ezrin has a role in cell spreading and motility. A notable feature of PACE-1 is the presence of a putative N-terminal kinase domain; however, in biochemical assays PACE-1 was shown to have associated rather than intrinsic kinase activity. Together these data suggest that PACE-1 may play a role in regulating cell adhesion/migration complexes in migrating cells. PMID- 12651156 TI - PINCH2 is a new five LIM domain protein, homologous to PINCHand localized to focal adhesions. AB - PINCH is a five LIM domain protein involved in the regulation of integrin mediated cell adhesion. It has been shown that PINCH interacts with integrin linked kinase and Nck2. Here we describe a new isoform of PINCH, which we call PINCH2. Therefore, we rename PINCH to PINCH1. PINCH2 has an overall similarity of 92% to PINCH1 and contains five LIM domains like PINCH1. While protein and gene structure of the PINCH homologues are very similar and well conserved during evolution, we observed differential expression pattern of the mRNAs. Based on northern hybridization of mouse embryo RNA, PINCH1 is already detectable at E8.5. It is highly expressed during later stages of development and in all adult mouse tissues analyzed, with the highest levels in heart, lung, bladder, skin, and uterus. In contrast, significant PINCH2 expression starts at E14.5. In adult mice it is widely expressed, similar to PINCH1, but absent from spleen and thymus. In situ hybridization confirmed the Northern data and showed differential expression of PINCH1 and PINCH2 in embryonic intestine. Finally, we demonstrate that PINCH2 localizes to focal adhesions in NIH 3T3 cells and to Z-disks in primary rat cardiomyocytes. PMID- 12651157 TI - Identification of CHE-13, a novel intraflagellar transport protein required for cilia formation. AB - Cilia are present on cells of many eukaryotic organisms and recent data in the mouse suggest that ciliary defects can cause severe developmental abnormalities and disease. Studies across eukaryotic systems indicate that cilia are constructed and maintained through a highly conserved process termed intraflagellar transport (IFT), for which many of the proteins involved have yet to be identified. IFT describes the movement of large protein particles consisting of an A and a B complex along the cilia axoneme in anterograde and retrograde directions. Herein we describe a novel C. elegans gene, F59C6.7/9, that is required for cilia assembly and whose function is disrupted in che-13 ciliogenic mutants. As previously shown for all IFT complex B genes identified to date, expression of che-13 (F59C6.7/9) is regulated by the RFX-type transcription factor DAF-19, suggesting a conserved transcriptional pathway in ciliogenesis. Fluorescent-tagged CHE-13 protein concentrates at the base of cilia and moves along the axoneme as expected for an IFT protein. Furthermore, loss of che-13 differentially affects the localization of two known IFT complex B proteins, OSM 5 and OSM-6, implying that CHE-13 functions as part of this complex. Overall, our data confirm that CHE-13 is an IFT protein and further that the IFT particle assembles in an ordered process through specific protein-protein interactions. PMID- 12651158 TI - Prolonged progestin treatment induces the promoter of CDK inhibitor p21 Cip1,Waf1 through activation of p53 in human breast and endometrial tumor cells. AB - Progestins are frequently used in the treatment of advanced breast and endometrial cancer. The human breast carcinoma cell line T47D shows a biphasic response to progestins. Short-term progestin treatment leads to enhanced DNA synthesis, while this line is growth inhibited upon prolonged exposure. An important protein involved in growth regulation by progestins in this cell is the CDK inhibitor p21(Cip1,Waf1). We show that after 1 day of progestin treatment in T47D cells, the p21 promoter-proximal region containing Sp1 binding sites is crucial in the induction by progestins. However, after 3 days the activity of the promoter-distal region becomes predominant in T47D cells or the endometrial carcinoma cell line ECC1. This is dependent upon two domains within this region that contain p53 response elements. In ECC1 and T47D cells 3-day progestin treatment induces a reporter containing a p53 response element, but not a mutated version. This induction is due to activation of p53 by progestin, which may be caused by nuclear translocation of p53. These data indicate that upon prolonged exposure, progestins activate p53, in human breast and endometrial tumor cells, which up-regulates the p21(Cip1,Waf1) promoter. This may be an important mechanism involved in progestin-inhibited cellular proliferation in these cells. PMID- 12651159 TI - Nucleus alignment and cell signaling in fibroblasts: response to a micro-grooved topography. AB - Cellular response to scaffold materials is of great importance in cellular and tissue engineering, and it is perhaps the initial cell contact with the scaffold that determines development of new tissue. Material surface morphology has strong effects on cell cytoskeleton and morphology, and it is thought that cells may react to the topography of collagen and surrounding cells during tissue embryology. A poorly understood area is, however, gene-level responses to topography. Thus, this paper used microarray to probe for consistent gene changes in response to lithographically produced topography (12.5 x 2-microm grooves) with time. The results showed many initial gene changes and also down-regulation of gene response with time. Cell and nucleus morphology were also considered, with nuclear deformation linked to cell signaling. PMID- 12651160 TI - Compartmentalization of Fas and Fas ligand may prevent auto- or paracrine apoptosis in epithelial cells. AB - Oligomerization of Fas receptor by its ligand, FasL, activates a signaling cascade that leads to apoptosis of Fas bearing cells. Interestingly, many epithelia coexpress Fas and FasL, yet FasL does not trigger Fas present on the same or neighboring cells to induce spontaneous apoptosis. Here, we show that Fas and FasL are segregated from each other to different cellular compartments in kidney epithelial MDCK cells. While Fas is restricted to the basolateral surface, FasL is sequestered to an intracellular compartment and, a lesser extent, the apical surface. This spatial segregation of Fas and FasL may explain how epithelial cells can constitutively express a functional Fas pathway but avoid auto- or paracrine cell death. Compromising this spatial segregation in physiological or pathological situations may play a so far underestimated role in initiating apoptosis of epithelial cells. PMID- 12651161 TI - Roles of mitogen-activated protein kinase and phosphoinositide 3'-kinase in ErbB2/ErbB3 coreceptor-mediated heregulin signaling. AB - ErbB2/HER2 and ErbB3/HER3, two members of the ErbB/HER family, together constitute a heregulin coreceptor complex that elicits a potent mitogenic and transforming signal. Among known intracellular effectors of the ErbB2/ErbB3 heregulin coreceptor are mitogen-activated protein kinase (MAPK) and phosphoinositide (PI) 3-kinase. Activation of the distinct MAPK and PI 3-kinase signaling pathways by the ErbB2/ErbB3 coreceptor in response to heregulin and their relative contributions to the mitogenic and transformation potentials of the activated coreceptor were investigated here. To this end, cDNAs encoding the wild-type ErbB3 protein (ErbB3-WT) and ErbB3 proteins with amino acid substitutions in either the Shc-binding site (ErbB3-Y1325F), the six putative PI 3-kinase-binding sites (ErbB3-6F), or both (ErbB3-7F) were generated and expressed in NIH-3T3 cells to form functional ErbB2/ErbB3 heregulin coreceptors. While the coreceptor incorporating ErbB3-WT activated both the MAPK and the PI 3 kinase signaling pathways, those incorporating ErbB3-Y1325F or ErbB3-6F activated either PI 3-kinase or MAPK, respectively. The ErbB2/ErbB3-7F coreceptor activated neither. Elimination of either signaling pathway lowered basal and eliminated heregulin-dependent expression of cyclin D1, which was in each case accompanied by an attenuated mitogenic response. Selective elimination of the PI 3-kinase pathway severely impaired the ability of heregulin to transform cells expressing the coreceptor, whereas attenuation of the MAPK pathway had a lesser effect. Thus, while both pathways contributed in a roughly additive manner to the mitogenic response elicited by the activated ErbB2/ErbB3 coreceptor, the PI 3 kinase pathway predominated in the induction of cellular transformation. PMID- 12651162 TI - Regulation of trespin expression by modulators of cell growth, differentiation, and apoptosis in prostatic epithelial cells. AB - We recently identified a novel rat ov-serpin, Trespin, which inhibits the trypsin like serine proteinase plasmin and is expressed in several tissues, including prostate. In this report Trespin expression was studied in prostatic cell lines, NRP-152, NRP-154, and DP-153, derived from the Lobund-Wistar rat. Northern blots revealed Trespin mRNA is expressed in NRP-152 and DP-153 basal epithelial cell lines but not in the luminal line, NRP-154. Similarly, Trespin levels drop >30 fold following transdifferentiation of NRP-152 cells toward a luminal variant, further suggesting Trespin expression is specific for basal prostatic epithelial cells. Trespin expression in NRP-152 cells is up-regulated by dexamethasone (Dex) and insulin-like growth factor-I (IGF-I), each of which stimulate growth and prevent differentiation and apoptosis. However, Dex (alone) facilitates loss of Trespin by TGF-beta, yet enhances the ability of LR(3)-IGF-I to reverse such loss, similar to the pattern of apoptosis induced by TGF-beta. Likewise, several apoptosis inducers markedly decrease Trespin mRNA levels. HEK293 cells stably overexpressing Trespin display increased cell proliferation and partial resistance to growth inhibition and phosphorylation of c-Jun induced by the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA). Together these data strongly suggest that Trespin has critical functions tied to the regulation of growth, differentiation, and apoptosis of prostatic epithelial cells. PMID- 12651164 TI - New initiatives in phlebology. PMID- 12651163 TI - UVB-induced apoptosis in normal human keratinocytes: role of the erbB receptor family. AB - Exposure of human keratinocytes to ultraviolet B (UVB) light leads to the activation of a variety of cell-surface receptors; however, the biologic consequences of these activated receptors are still unclear. It was previously reported that inhibition of cellular tyrosine kinase activity suppressed UVB dependent effects in human skin. We confirmed that the same suppression of UVB induced apoptosis occurs in normal human keratinocytes grown in culture. Furthermore, we sought to determine the role of erbB receptor tyrosine kinases in human keratinocytes following UVB irradiation. Using a specific inhibitor of the erbB family of tyrosine kinase receptors, DAPH, we investigated the effects of UVB-dependent activation of these receptors on keratinocyte biology. The addition of DAPH to keratinocytes resulted in the concentration-dependent protection of UVB-induced apoptosis. The protection from apoptosis was not due to the induction of keratinocyte differentiation, the loss of keratinocyte viability, or inhibition of the proliferative potential of keratinocytes by DAPH. The effect of DAPH on apoptosis was specific for UVB as it had no effect on bleomycin-induced apoptosis. Furthermore, the inhibition of UVB-induced apoptosis could also be observed using neutralizing antibodies to either erbB1 or erbB2. Finally, we demonstrated that DAPH could also inhibit UVB-induced apoptosis in an epidermal organotypic model system. These studies suggest an important role for the erbB receptors in UVB-induced apoptosis of human keratinocytes. PMID- 12651165 TI - Improving the patency of vascular bypass grafts: the role of suture materials and surgical techniques on reducing anastomotic compliance mismatch. AB - BACKGROUND: compliance mismatch is an important factor in the development of myointimal hyperplasia in both coronary and vascular anastomoses. This mismatch may be reduced by the use of newer suture materials and techniques. This review discusses the current techniques and materials used to date in generating anastomoses in both coronary and vascular applications and to correlate these with the degree of inherent compliance achieved. METHODS: PubMed, ISIS, CAS and PAS database searches were performed. Other articles were cross-referenced. RESULTS AND CONCLUSION: continuous suture is still the most used technique in both cardiac and vascular surgery for the generation of anastomoses due to the reduced time and improved haemostasis. However, continuous suture results in a greater compliance mismatch than the interrupted technique. Vein cuffs and patches improve compliance and transmission of pulsatile blood flow and offer improvement of graft patency. Alternative to sutures are biological glue, clips and laser generated solders all of which have shown promising results, but further work is required before they become applicable for routine use. PMID- 12651166 TI - Radiocontrast induced nephropathy. AB - In spite of improvements in chemical structure, contrast media assisted X-ray examination is still the third leading cause of hospital-acquired acute renal failure. An increase >50% or >88 micro mol/L in S-creatinine is a clinically important acute renal failure. The peak in S-creatinine occurs within 2-5 days after exposure. The frequency of oliguria, transient or permanent haemodialysis is unknown. The cause is a hypoxic tubular injury due to vasoconstriction with release of free oxygen radicals. Major risk factors are prior renal insufficiency and diabetes mellitus. Minor risk factors are congestive heart disease, dehydration, hypotension, hypoxia, amount of contrast, ionic and high osmolar contrast, repeated examinations at short intervals, abdominal examination, and perhaps age, smoking, hypercholesterolaemia, and use of Non-Steroidal Anti inflammatory Drug. Prevention seems possible by omission or reduction of contrast, ameliorating predisposing factors, saline hydration 24h before and after exposure, and 600 mg acetylcysteine orally twice daily 24h before and after exposure. A three-day treatment with 20mg nitrendipine daily, starting 1 day before examination may also be preventive. The present research is unfortunately characterised by small numbers, lack of clinical important renal failure, and lack of long term results. The latter may be important after new data indicate that radiation may trigger a chronic oxidative process through a similar pathway. PMID- 12651167 TI - Tissue-engineered bioprosthetic venous valve: a long-term study in sheep. AB - OBJECTIVE: to develop a graft bearing an immunologically tolerated tissue engineered venous valve (TE graft) that will be incorporated into a native vessel, and restore normal valve function for the treatment of chronic venous insufficiency. METHODS: twenty-four TE grafts were grown using decellularised allogeneic ovine veins as donor matrix, which was subsequently repopulated with the future recipient's myofibroblasts (MFB) and endothelial cells (EC). TE grafts were implanted into the external jugular vein. Animals were sacrificed at 1, 6, and 12 weeks (n=4, each). Autografts served as controls (1 week, n=4; 6 weeks, n=4). Specimen for histology and immunohistochemistry were taken. RESULTS: the matrix was successfully repopulated with MFB and EC (n=8). Patency on venography in the TE graft-group was44,44, and 34 at 1, 6, and 12 weeks, and44 (44) in autografts at 1 (6) weeks, respectively. Except for 2 TE grafts after 12 weeks, valves were competent (duplex ultrasound). Patent TE grafts were merely distinguishable from autografts with minor inflammatory reactions. Reflux was caused by neo-intima formation related to the basis of the TE graft. CONCLUSION: acellularisation and consecutive in vitro autogeneic re-seeding of valved venous conduits can lead to immunologically acceptable, patent, and competent implants in sheep. PMID- 12651168 TI - Minimally invasive surgical management of primary venous ulcers vs. compression treatment: a randomized clinical trial. AB - OBJECTIVES: to compare minimally invasive surgical haemodynamic correction of reflux (CHIVA) with compression in the treatment of venous ulceration. DESIGN: prospective randomised study. MATERIALS AND METHODS: from a cohort of 80 patients with 87 venous leg ulcers, 47 were randomised to either surgery or compression. RESULTS: at a mean follow-up of 3 years, healing was 100% (31 days) in the surgical and 96% (63 days), in the compression group (p<0.02). The recurrence rate was 9% in the surgical and 38% in the compression group (p<0.05). In the surgical group, all plethysmographic parameters except ejection fraction, had improved significantly at 6 months in the surgical group, and at 3 years residual volume fraction remained in the normal range. Finally, quality of life significantly improved in the operated group. CONCLUSIONS: this study supports the effectiveness of surgical therapy for leg ulceration secondary to superficial venous reflux. PMID- 12651169 TI - Pathogenesis of varicose veins. AB - BACKGROUND: varicose veins are present in up to 40% of the population. They may be associated with considerable morbidity. Around 70% of patients with chronic venous insufficiency have evidence of superficial venous incompetence on duplex imaging. METHODS: in this publication we review the evidence available regarding the pathogenesis of varicose veins. RESULTS: a number of theories have been suggested. The present data suggests that abnormalities in the venous endothelium and smooth muscle cells result in vein wall dilatation with secondary valvular incompetence. However, there is also evidence to support acquired venous valve failure. Congenital venous valve abnormalities have not been well studied. CONCLUSION: further work is required on this relatively neglected topic. The use of improved imaging such as high-resolution ultrasound is likely to significantly improve our understanding of venous valve function and pathology. PMID- 12651170 TI - Improving endothelial cell retention for single stage seeding of prosthetic grafts: use of polymer sequences of arginine-glycine-aspartate. AB - OBJECTIVE: single stage seeding within the timeframe of a typical vascular operation has not been successful. One reason for this is poor cell adherence to the graft lumen once exposed to pulsatile blood flow. In this study we have carried out investigations with the use of two different fibronectin-based peptides, fibronectin-like engineered protein polymer (FEPP) which contains multiple copies of arginine-glycine-aspartate (RGD) and fibronectin adhesion promoting peptide (FAPP) to improve cell adherence. MATERIALS AND METHODS: FAPP and FEPP were coated onto native polyurethane and heparinised polyurethane grafts. The grafts were then seeded for either 1 or 2h with human umbilical vein endothelial cells (HUVEC). After the incubation period the cells were washed off and cell retention was calculated. Cell metabolism was measured using Alamar Blue, and confirmed with scanning electron microscopy (SEM). RESULTS: heparinised grafts coated with FEPP showed the best cell retention after both 1 and 2h seeding (80+/-4% vs 81+/-3%). This graft had no significant difference in cell retention after both times whilst all the other grafts had better cell retention after a 2h seeding. The Alamar blue and SEM results confirmed cell viability and function for all graft types. CONCLUSION: heparinised graft coated with FEPP allows significant cell retention after only 1h of seeding and shows promise for single stage seeding. PMID- 12651171 TI - The effect of lower limb ischaemia-reperfusion on intestinal permeability and the systemic inflammatory response. AB - OBJECTIVES: a relationship has been demonstrated between increased intestinal permeability, endotoxaemia and the development of the systemic inflammatory response syndrome (SIRS) after aortic surgery. The aim of this study was to evaluate whether isolated lower limb ischaemia-reperfusion (I/R) injury affects intestinal mucosal barrier function and cytokine release. PATIENTS AND METHODS: four groups of patients were investigated, group I, patients with critical limb ischaemia (CLI) undergoing infra-inguinal bypass surgery (n=18); group II, patients with intermittent claudication (IC) undergoing infra-inguinal bypass surgery (n=14); group III, patients with CLI unsuitable for arterial reconstruction, undergoing major amputation (n=12); and group IV, patients undergoing carotid endarterectomy for symptomatic carotid stenosis (n=13). Intestinal permeability, endotoxaemia and urinary soluble tumour necrosis factor receptors were assessed (p55TNF-R). RESULTS: an increase in intestinal permeability was observed on the 3rd postoperative day only in CLI group. This was found to correlate with arterial clamp time. Patients who had a femoro-distal bypass had significantly higher intestinal permeability compared to those who had femoro-popliteal bypass. Endotoxaemia was not detected in any of the groups. Postoperative urinary p55TNF-R concentrations were significantly higher in CLI group compared to the other groups. These did not correlate with the increased intestinal permeability. CONCLUSIONS: our results support the hypothesis that revascularisation of critically ischaemia limbs leads to intestinal mucosal barrier dysfunction and cytokine release. They also suggest that the magnitude of the inflammatory response following I/R injury is related to the degree of initial ischaemia. PMID- 12651172 TI - Differentiation of primary and secondary Raynaud's disease by carotid arterial stiffness. AB - INTRODUCTION: primary Raynaud's disease may be difficult to differentiate clinically from the secondary form with an underlying connective tissue, haematological, neurovascular or drug-induced disorder. We undertook a study to determine the elastic carotid and muscular femoral arterial biomechanical properties and intima-media thickness (IMT) in subjects with primary and secondary Raynaud's disease, to assess whether these parameters could differentiate the two conditions. METHODS: twenty patients with primary Raynaud's disease and 53 subjects with secondary Raynaud's associated with scleroderma (systemic sclerosis, SSc) had measurements of their carotid and femoral wall mechanics with a duplex scanner coupled to a Wall Track system. Their age, gender, body mass index, heart rate, systolic and diastolic blood pressures, presumed cardiovascular load, plasma creatinine, fasting cholesterol, triglyceride and glucose concentrations were also measured. RESULTS: the carotid elastic properties [mean (SD): elastic modulus: 560 (180) vs 1204 (558) mmHg,p <0.001 and stiffness index: 5.69 (1.35) vs 11.92 (6.4), p<0.001 for primary and secondary Raynaud's respectively] were significantly impaired in patients with secondary Raynaud's disease even after adjustment for potentially influencing physiological and biochemical variables. There were no statistical differences in the femoral elastic properties or the carotid and femoral IMTs between the two groups. CONCLUSION: Duplex determination of the carotid elasticity or stiffness is different in primary Raynaud's phenomenon compared with secondary Raynaud's associated with SSc. This may be a useful non-invasive tool, in addition to autoantibody markers and nail-fold capillaroscopy, to differentiate between the two forms of Raynaud's phenomenon. PMID- 12651173 TI - Outcome of surgery for vascular access in patients commencing haemodialysis. AB - OBJECTIVE: assessment of surgical vascular access procedures for haemodialysis. DESIGN: retrospective cohort audit. MATERIALS AND METHODS: secondary patency was calculated from surgery until access failure, death, transplant, conversion to peritoneal dialysis, or loss to follow-up. All surgical procedures including immediate failures and failures to mature fistulae were included but not radiological interventions. RESULTS: four hundred and forty-five operations were undertaken in 197 patients over 87 months comprising 273 access creations and 172 revisions. Median follow-up was 26 months with a mortality of 9.4 deaths per 100 patient-years including eight perioperative deaths. Autogenous access was created in 147 (75%) patients with 142 based on the radial artery whilst 50 prosthetic grafts including 46 PTFE grafts and 40 forearm loops were placed. Patients receiving grafts were more likely to be older, female and die in follow-up. Grafts had higher patencies of 89, 75 and 68% at 1, 2 and 4 years, respectively compared to 69, 63 and 55% for autogenous access. This difference was significant (p=0.049) when the effects of the presence of diabetes and peripheral arterial disease were accounted for but more frequent revisions were required. The final access placed was autogenous in 110 (56%) and prosthetic in 87 (44%) patients. CONCLUSIONS: in our surgical unit, there was high secondary patency including for prosthetic grafts, high autogenous utilisation and relatively infrequent reintervention. PMID- 12651174 TI - Long-term survival and health-related quality of life of patients with ruptured abdominal aortic aneurysm. AB - INTRODUCTION: the outcome of ruptured abdominal aortic aneurysm (RAAA) patients is most frequently measured as operative or in-hospital mortality rate. However, survival alone is not an indicator of quality of the treatment. Assessment of quality of life (QoL) is used increasingly and is a relevant measure of outcome. OBJECTIVE: to assess long-term survival and QoL of patients undergoing repair of RAAA. DESIGN: follow-up study with cross-sectional QoL evaluation. MATERIALS AND METHODS: between 1996 and 2000, 199 of 220 patients with RAAA underwent surgery. Survivors were sent the generic the RAND 36-item Health Survey (RAND-36) self administered questionnaire. RESULTS: total hospital mortality and operative mortality were 103 of 220 (47%) and 82 of 199 (41%). Of the 117 initial survivors, 21 were deceased at the time of the study. When compared to an age- and sex-adjusted general population, only physical functioning was significantly impaired (p=0.01) in the 82 of 93 (88%) RAAA survivors who responded. CONCLUSIONS: survivors after repair of RAAA had almost the same QoL as the norms of an age- and sex-adjusted general population, justifies an aggressive operative policy in RAAA. PMID- 12651175 TI - Aortic side branch perfusion alone does not account for high intra-sac pressure after endovascular repair (EVAR) in the absence of graft-related endoleak. AB - OBJECTIVE: to examine the effect of stent-graft deployment on pressure within an aneurysm sac and to investigate the potential sources of intra-sac pressure. MATERIAL AND METHODS: intra-sac pressure was monitored during and immediately after endovascular repair via an indwelling catheter. Intra-sac pressure was also monitored during conventional open repair and was compared with the pressure measured within patent lumbar and inferior mesenteric side-branches, both before and after restoration of iliac arterial blood flow. Intra-sac and side-branch pressures were recorded and expressed as ratios of simultaneously measured radial artery pressure. RESULTS: in the absence of a graft-related endoleak (23/25 patients), endovascular repair resulted in a significant reduction in intra-sac pulse pressure (median ratio 0.31 IQR 0.10-0.46). There was no corresponding reduction in mean intra-sac pressure (median ratio 0.91; IQR 0.83-1.00). Application of clamps at conventional open repair resulted in a fall in both intra-sac pressure (median ratio 0.39, IQR 0.32-0.64) and pressure within side branches (median ratio 0.45, IQR 0.33-0.64). Restoration of iliac blood flow resulted in a modest recovery of the side-branch pressure (median ratio 0.63, IQR 0.57-0.81), which nonetheless remained significantly less than the intra-sac pressure recorded after EVAR (p=0.01). CONCLUSION: reperfusion of the aneurysm sac through patent side-branches seems insufficient to account for persistent pressurisation of the aneurysm after endovascular repair. This finding supports the hypothesis that pressure may be transmitted directly through stent-graft fabric. PMID- 12651176 TI - Aneurysm of an expanded polytetrafluoroethylene vascular graft: an ultrastructural evaluation. PMID- 12651178 TI - Aortic surgery through a transverse mini-laparotomy. PMID- 12651177 TI - How many claudicants should be prescribed statins? PMID- 12651180 TI - Characterization of a small plasmid (pMBCP) from bovine Pseudomonas pickettii that confers cadmium resistance. AB - This is the first report of isolation of Pseudomonas pickettii from a normal adult bovine duodenum. This organism was one of several bacteria isolated as part of a study to examine cadmium resistance genes (cad(r)) for use in generating transgenic plants to reclaim cadmium-contaminated soils in Kansas. P. pickettii containing a plasmid of 2.2kb (designated pMBCP) grew in Luria-Bertani broth and agar containing up to 800 microM of cadmium chloride and was resistant to 16 antibiotics. Curing the organism of plasmid revealed that antibiotic resistances were not plasmid-mediated. Low-level cadmium resistance was conferred by the plasmid because uncured organism grew significantly better (P<0.05) at 55 microM compared to cured organism. Both plasmid and chromosomal DNA were probed by DNA DNA hybridization for the presence of known cadmium resistance genes (cadA, cadC, and cadD from Gram-positive (Staphylococcus aureus), but none were detected. The plasmid had one restriction site each for BamHI, PstI, SmaI, and XhoI; two sites each for HincII, SacI, and SphI; and multiple sites for AluI and XcmI. DNA sequence analyses of the cloned and original plasmids showed a GC content of greater than 60% and no homology to any published sequences in the GenBank, European Bioinformatics Institute, or Japanese Genome Net databases. The DNA sequence is contained in GenBank accession number AF144733. Thus, pMBCP offers low-level cadmium resistance to P. picketttii. PMID- 12651181 TI - Cesium-137 concentrations in marine macroalgae from different biotopes in the Aegean Sea (Greece). AB - The commonest species of red, brown, and green macroalgae were sampled from a range of biotopes in the Aegean Sea during 1998 for their 137Cs content. Significant differences in 137Cs concentrations were found among different species growing under similar environmental conditions, suggesting that uptake does not follow physical levels but is influenced by allometric parameters and physiological mechanisms. 137Cs content in collected seaweeds were in the descending order: Ulva lactuca>Enteromorpha linza>Gracilaria verrucosa>Ceramium rubrum>Cystoseira barbata. A comparison of the stations studied indicated that the degree of accumulation is also dependent on the geomorphology of the specific area. The west side of Thermaikos Gulf, where the rivers Axios, Aliakmon, and Pinios flow, was highly contaminated as was the Strymonicos Gulf where the river Strymon flows. High cesium concentrations were also observed in algae from Lemnos Island near the mouth of the Dardanelles Channel, most likely due to the horizontal circulation of water masses from the Marmaras Sea to the Aegean Sea resulting in the discharge of 137Cs during the purification process in the Black Sea through the Dardanelles Straits. It is concluded that the general dispersion of 137Cs results from the water circulation regime within the Aegean Sea. Higher levels are present in the northern part of the basin due to current circulation originating from the Black Sea and from outflow of rivers. Levels decrease to the south and east, but increase again to the west of Crete where the Black Sea water mass can be periodically traced. PMID- 12651182 TI - Reproduction, blood and plasma parameters and gill histology of vendace (Coregonus albula L.) in long-term exposure to acidity and aluminum. AB - Vendace were exposed to pHs 4.75 and 5.25 with or without added aluminum (200 microg=7.4 micromol AlL(-1)) from late endogenous vitellogenesis in July through the spawning period. At the normal time of spawning, when 48% of the control females had already released their eggs, 50% of females at pH 4.75+Al had completely unovulated oocytes. The final proportions of completely ovulated females were 14%, 36%, 25%, 61%, and 81% at pH 4.75+Al, pH 4.75, pH 5.25+Al, pH 5.25, and in the control group, respectively. Delayed testes regression was seen in males at pH 4.75+Al. A clear decrease in plasma Na(+) and Cl(-) and an increase in blood glucose concentration was detected only near spawning time, from October to November, coincident with Al accumulation inside the gill tissue. It is concluded that seasonal changes, probably related to reproductive physiology or to the decrease in water temperature, are associated with the increase in Al toxicity in vendace. PMID- 12651183 TI - Metal uptake from soils and soil-sediment mixtures by larvae of Tenebrio molitor (L.) (Coleoptera). AB - Bioassays were performed to evaluate the impact of soil characteristics on Cd, Cu, Pb, and Zn uptake by larvae of Tenebrio molitor. Metal accumulation was determined in 13 natural field soils, one metal-spiked field soil, four soil sediment mixtures, and Cd- or Zn-spiked OECD artificial soil. Statistical analyses were used to investigate covariation of accumulation patterns with various soil metal pools and soil properties. Body concentrations of Cu and Zn in Zn-spiked OECD soils, field soils, and soil-sediment mixtures mostly remained constant. Considerable variation was noted for all Cd and Pb steady-state body concentrations among field soils and soil-sediment mixtures. For the spiked field soil and in the Cd-spiked OECD soil, body concentrations increased almost linearly with time. For the nonessential metals Cd and Pb, larval body concentrations correlated mainly to the total metal pool of the soil. Cd uptake at similar total Cd concentrations was within the same range among spiked OECD soils, field soils, and mixtures. A comparison of the findings with studies on other soil-inhabiting species shows that metal uptake patterns depend on metal type, soil type, and exposed species. It is suggested that soil organisms can be categorized according to gross divergence in ecophysiological characteristics, determined by, for instance, (non)permeability of the outer integument. These characteristics appear as similarities among multivariate functions as derived for the beetle. PMID- 12651184 TI - Sediment toxicity tests using benthic marine microalgae Cylindrotheca closterium (Ehremberg) Lewin and Reimann (Bacillariophyceae). AB - A new method for sediment toxicity testing using marine benthic pennate noncolonial diatom (Cylindrotheca closterium, formerly Nitzschia closterium) has been developed. This microalgae showed a good growth rate during the experimental period, even when low enriched media were used. Sediment spiked with heavy metals [cadmium (Cd), copper (Cu), and lead (Pb)] was employed to determine the EC(50) values, using microalgal growth inhibition as the endpoint. The obtained results were as follows: Three heavy metals (Cd, Cu, and Pb), previously spiked on experimental sediment, were separately assayed in toxicity tests. The EC(50) values for these heavy metals in microalgal growth inhibition tests resulted to be 79 mg kg(-1) for Cd, 26 mg kg(-1) for Cu, and 29 mg kg(-1) for Pb (in experimental sediment). The influence of sediment granulometry on the growth of microalgal population was also studied, finding that the growth of the microalgal population on media containing sediment with a relation sand-size:silt size of 9:1 was not different from optimal growth (occurring in media containing 100% sand-sized sediment). The diatom C. closterium proved to be a suitable organism for sediment toxicity tests, due to its sensitivity and fast growth even in poorly enriched media. PMID- 12651185 TI - Toxic effects of chlorpyrifos on morphology and acetylcholinesterase activity in the earthworm, Eisenia foetida. AB - The acute toxicity of chlorpyrifos (O, O-diethyl-O-(3,5,6-trichloro-2-pyridyl) phosphorothioate) was determined in the earthworm, Eisenia foetida. A 48-h contact test as described by the Organization for Economic Cooperation and Development (OECD) guideline 207 was carried out. The LC(50) of chlorpyrifos was 0.063 microg/cm(2). Inhibition of acetylcholinesterase (AChE: EC 3.1.1.7) activity indicated by in vitro neurotoxic potentiality revealed competitive inhibition and altered K(m) values widely in a dose-dependent manner. The K(i) value of chlorpyrifos was 4.20 x 10(-6)M. AChE activity of LC(50)-exposed worms was 62%, 79%, 85%, and 91% inhibited at 12, 24, 36, and 48h, respectively. Scanning electron microscopic studies revealed the morphological abnormalities in the worms. The present study demonstrates a dose- and time-dependent exposure of chlorpyrifos through skin results, morphological abnormalities, and inhibition of AChE in the earthworm, E. foetida. PMID- 12651186 TI - Identification of endocrine-disrupting effects in aquatic vertebrates and invertebrates: report from the European IDEA project. AB - The EU-funded project IDEA aimed to evaluate (a) what parameters and endpoints allow the detection of endocrine-mediated developmental and reproductive effects of (xeno)estrogens in life cycle- and life stage-specific toxicity tests with the zebrafish Danio rerio, a small laboratory fish used in many ecotoxicity test guidelines, and (b) whether substances that act as estrogens in vertebrates may also adversely affect the development, differentiation, and reproduction of aquatic invertebrates. The invertebrate species investigated included Hydra vulgaris, Gammarus pulex, Chironomus riparius, Hyalella azteca, and Lymnaea stagnalis. The animals were exposed to the model estrogenic chemicals ethynylestradiol (EE2), bisphenol A (BPA), and octylphenol (OP), which exert their endocrine activity in vertebrates through the estrogen receptor. As endpoints, developmental and reproductive parameters at the organism level as well as molecular and cellular parameters were measured. Life cycle exposure of zebrafish to (xeno)estrogens induced a specific, partly irreversible response pattern, consisting mainly of (a) induction of vitellogenin (VTG), (b) alterations of gonad differentiation, (c) delay of first spawning, and (d) reduced fertilization success. The effects of EE2 on zebrafish were expressed at environmentally realistic concentrations, while BPA and OP became effective at concentrations higher than those usually found in the environment. The vitellogenic response was equally sensitive as the reproductive parameters in the case of EE2, but VTG was more sensitive in the case of BPA. Partial life cycle exposure of zebrafish had lasting effects on fish development and reproduction only when the fish were exposed during the stage of juvenile bisexual gonad differentiation. In (partial) life cycle and multigeneration studies with invertebrates, (xeno)estrogenic impact was assessed by a range of developmental and reproductive parameters including hatching, growth, moulting, mating behavior, and egg number. Several parameters were found to be responsive to (xeno)estrogens; however, most effects were induced only at higher, probably nonphysiological concentrations. Low-dose effects were observed in full life cycle experiments, particularly in the second generation. It remains to be established whether the estrogen-induced alterations in the invertebrate species indeed do result from disturbances of the endocrine system. The findings of the present research project support the development of appropriate testing methodologies for substances with estrogenic activity. PMID- 12651187 TI - Potencies of estrogenic compounds in in vitro screening assays and in life cycle tests with zebrafish in vivo. AB - The objective of this study was to compare the estrogenic potency of environmental estrogens at two testing tiers: at the initial level of in vitro screening assays, and at the level of definitive fish reproduction tests in vivo. The in vitro tests included a recombinant yeast estrogen receptor (ER) assay, a competitive radioreceptor assay using the hepatic ER of carp (Cyprinus carpio), and assays on vitellogenin induction in cultured hepatocytes of rainbow trout (Oncorhynchus mykiss) and carp. In vivo, full life cycle tests with zebrafish (Danio rerio) were performed, using fertilization success as estrogen-sensitive reproductive endpoint. The test compounds included the natural estrogen 17beta estradiol (E2) (only applied in the in vitro assays); the synthetic estrogen ethynylestradiol (EE2); and two xenoestrogens, 4-tert-octylphenol (OP) and bisphenol A (BPA). Among the in vitro assays, differences were observed in the relative ranking of the test substances, and in the absolute sensitivity (EC50 values), although the interassay differences of EC50 values were within one order of magnitude. The in vivo activity of the test compounds was not accurately predicted by the in vitro assays, with respect to neither sensitivity nor ranking. The in vitro assays tended to overestimate the relative potency of the xenoestrogens; i.e. the ratio between the activity of the reference compound, EE2, and that of the test compound. The best prediction of the in vivo fish test results was obtained from the recombinant yeast assay. PMID- 12651188 TI - Comet assay on gill cells and hemocytes from the blue mussel Mytilus edulis. AB - Gill cells and hemocytes from the blue mussel Mytilus edulis were examined for DNA damage using the comet assay after laboratory exposure in vitro and in vivo to methyl methansulfonate (MMS). Hydrogen peroxide and UV radiation were used as positive control. Comet assay was also carried out on hemocytes from blue mussels sampled at polluted and unpolluted coastal areas. After 60 min in vitro exposure of gill cells to MMS, the highest response, a tail moment of 6.70+/-4.25, was obtained at 1.0mg/L. At higher doses the response decreased. After 2 days in vivo exposure a dose response was seen at concentrations between 1.0 and 33.0mg/L MMS for both gill cells and hemocytes. However, after 4 days in vivo exposure using the same concentrations of MMS, a maximum effect was seen at a 10 times lower concentration of 3.3mg/L. At the higher doses, the effect decreased. Hemocytes from blue mussels sampled at four polluted sites in Koge Bay had a great variation in tail moments with the highest value of 5.38+/-4.39. The average of all samples from Koge Bay had tail moments of 2.75+/-1.00(n=19), which was significantly higher (P<0.05) than the average, 1.72+/-1.16(n=10), of samples from unpolluted coastal waters. PMID- 12651189 TI - Developmental, behavioral, and reproductive effects experienced by Japanese medaka (Oryzias latipes) in response to short-term exposure to endosulfan. AB - Japanese medaka (Oryzias latipes) were exposed to concentrations of 0.01, 0.10, and 1.0 microg/L of the pesticide endosulfan for 24h beginning either 4-6h postfertilization or 4-6h posthatch to determine effects on hatching time, growth, mobility, foraging ability, and reproduction. Eggs exposed to endosulfan took longer to hatch, and the resulting fry were smaller at 1 week of age and had decreased mobility at 2 weeks of age. Upon reaching sexual maturity, these individuals also produced fewer eggs, and these eggs took significantly longer to hatch. Medaka exposed to endosulfan shortly after hatching did not differ in early mobility or foraging ability. Interestingly, upon reaching sexual maturity, these individuals produced more eggs than did unexposed females. The observed effects were not dose-dependent, with medaka exposed to intermediate concentrations of endosulfan (0.10 microg/L) exhibiting the greatest response. These results suggest that short-term exposure to environmentally relevant concentrations of a common pesticide may have long-term effects on growth, behavior, and reproduction in fish. PMID- 12651190 TI - Gas exchange, growth, and chemical parameters in a native Atlantic forest tree species in polluted areas of Cubatao, Brazil. AB - The Atlantic forest species near the industrial complex of Cubatao, Brazil have been subjected to heavy air pollution for decades. In this study, we used some physiological parameters (gas exchange, growth and chemical contents) to biomonitor the effects of air pollution on Tibouchina pulchra, one of the most common tree species in this forest. Under standardized conditions, saplings were exposed to the environment from April to July and from July to September of 1998, at three different sites in the vicinity of the industrial complex: the Valley of Piloes River (VP), the control area; the Valley of Mogi River (VM), near fertilizer, metallurgical, and cement industries sustaining high concentrations of fluorides, N and S oxides, and particulate materials; and Caminho do Mar (CM), near petrochemical industries under N and S oxides, photooxidants, and organic compounds. Plants exposed to CM and VM conditions presented visible injuries, reductions in net photosynthesis, growth parameters, and ascorbate concentrations, and increased F, N, and S foliar concentrations. These results indicate that the environmental conditions around these industries are still harmful to plants. PMID- 12651191 TI - Comparative studies on algal toxicity testing using fluorometric microplate and Erlenmeyer flask growth-inhibition assays. AB - Two fluorometric microplate algae growth-inhibition assays with a liquid volume of 2 mL and 200 microL per well are presented, and comparative studies on the toxicity of chemicals are carried out with Erlenmeyer flask assays. The test procedures are in accordance with the standards ISO 8692 (DIN 38412 L9 and EN 28692), OECD 201 and DIN 38412 L33. By testing four toxicants several times laboratory internal repeatability is proven. Statistical evaluation demonstrates that the results obtained with both the 24-well and the 96-well microplates are nearly identical with the results of the Erlenmeyer flask assay. Therefore, the microplate growth-inhibition assays can be applied for the testing of a wide range of chemicals and environmental samples if some methodical aspects are taken into account. Apart from that, there is a strong need for harmonization if the dependency of the EC values on the toxicological endpoint is considered. PMID- 12651192 TI - Predicting petroleum phototoxicity. AB - Phototoxicity to Daphnia magna was studied on 14 polycyclic aromatic hydrocarbons (PAHs) and 22 petroleum products (ranging from diesel to crude oil). The phototoxicity ranking of pure PAHs was about the same as found in another study on fish gill cells in vitro (Toxicology 127 (1998) 143), suggesting that the relative acute phototoxicity does not differ significantly between different species. Most petroleum products were found to be phototoxic, although the results differ somewhat between test methods (preparation of water-accommodated fractions or petroleum ether dissolved oil slurries). The degree of phototoxicity of a sample is related to both the source and the refining process of the crude oil. The best chemical predictors found were the concentrations of eight phototoxic parent PAHs. An even simpler analytical technique suggested for initial screening would be direct measurements of "total PAH," where samples having more than 3% PAH should be studied further. PMID- 12651193 TI - Invertebrate biomarkers: links to toxicosis that predict population decline. AB - The application of biochemical measurements that can be used as individual biomarkers of impaired biological function in invertebrates is reviewed to evaluate whether biochemical biomarkers of aquatic invertebrates can predict changes in natural populations. Biomarkers that measure toxic effects at the molecular level (e.g., the inhibition of brain acetylcholinesterase activity by organophosphorus pesticides) have been shown to provide rapid quantitative predictions of a toxic effect upon individuals in laboratory studies. Such biomarkers should not be used as a replacement for conventional aquatic monitoring techniques, but should be applied as supplementary approaches for demonstrating links between sublethal biochemical and adverse effects in natural populations in field studies. The research challenge for using biomarker measurements in aquatic invertebrates is to predict effects at the population level from effects at the individual level measured upon individuals collected in the field. PMID- 12651194 TI - The best of times...the worst of times. PMID- 12651195 TI - Clinical utility of biochemical markers in colorectal cancer: European Group on Tumour Markers (EGTM) guidelines. AB - In recent years, numerous serum and cell/tissue-based markers have been described for colorectal cancer (CRC). The aim of this article was to provide guidelines for the routine clinical use of some of these markers. Lack of sensitivity and specificity preclude the use of any available serum markers such as carcinoembryonic antigen (CEA), CA 19-9, CA 242, CA 72-4, tissue polypeptide antigen (TPA) or tissue polypeptide-specific antigen (TPS) for the early detection of CRC. However, preoperative measurement of CEA is desirable as this may give independent prognostic information, help with surgical management and provide a baseline level for subsequent determinations. For patients with stage 2 (Dukes' B) and 3 (Dukes' C) disease who may be candidates for liver resection, CEA levels should be measured every 2-3 months for at least 3 years after diagnosis. For monitoring treatment of advanced disease, CEA should also be tested every 2-3 months. Insufficient evidence is presently available to recommend the routine use of other serum markers for monitoring purposes. Similarly, the new cell and tissue-based markers (e.g, ras, P53) cannot yet be recommended for routine clinical use. PMID- 12651196 TI - Potential role of magnetic resonance spectroscopy in assessment of tumour response in childhood cancer. AB - This brief review considers to what extent Magnetic Resonance Spectroscopy (MRS) can play a role in monitoring early tumour response with examples of preclinical studies and selected clinical studies in tumours of children and young adults. An early non-invasive indicator of tumour response to therapy would provide useful information regarding the effectiveness of therapy. This might be a relevant prognostic factor in new patients and in phase II studies could facilitate recommendations at an early stage as to whether to continue treatment. This review suggests that several markers and ratios are emerging as potential prognostic markers, but larger prospective studies are needed before translating this into clinical practice. PMID- 12651197 TI - Pregnancy after breast cancer. AB - The issue of pregnancy in patients previously treated for breast cancer is controversial. This paper reviews the literature using Medline and Embase databases over the last 50 years to address the issue. Overall survival in patients treated for breast cancer who subsequently become pregnant compares favourably with controls. This paper also addresses the effects of adjuvant therapy (loco-regional and systemic) on subsequent pregnancy. Introduction of a national registry of these patients may help inform such patients in the future. PMID- 12651198 TI - An EORTC-ECSG phase I study of LU 79553 administered every 21 or 42 days in patients with solid tumours. AB - A single-agent dose-escalating phase I and pharmacokinetic study on the naphthalamide agent, LU 79553, was performed to determine its safety profile, maximum tolerated dose (MTD) and recommended dose for phase II studies. LU 79553 was given intravenously (i.v.) every 3 weeks to patients with advanced solid cancers (an extended cohort of patients also received the drug every 6 weeks). 59 patients were enrolled into the study (50 patients in the 3-weekly schedule and 9 patients in the 6-weekly schedule). Dose levels studied ranged from 10 mg/m(2) to 160 mg/m(2). Neuro-muscular toxicity was identified as the dose-limiting toxicity (DLT). This muscular toxicity was observed after administrating total doses of 160-450 mg/m(2) (median 330 mg/m(2)). Non-DLTs consisted of diarrhoea, nausea and vomiting, fatigue and local venous phlebitis. The major haematological toxicities observed were anaemia and neutropenia (and were mainly observed at the two highest dose levels). The proposed dose for phase II studies using the 3-weekly regimen is 100 mg/m(2)/course (60 min infusion in 500 ml normal saline), but a close clinical follow-up of the patients for neuromuscular toxicity is mandatory. Prolongation of the treatment interval to 6 weeks, based upon the long half-life of the drug in the plasma and tissue, observed during this study, seemed not to be feasible in this heavily pretreated group of patients. PMID- 12651199 TI - An economic evaluation of the optimal workload in treating surgical patients in a breast unit. AB - A breast unit is a cancer centre specialised in the diagnosis and treatment of patients with breast cancer. The high level of specialised skills involved in running a breast unit makes it an expensive pattern of care. The European Society of Mastology (EUSOMA) recommends a minimum caseload of 150 cases sufficient to maintain expertise for each team member and to ensure cost-effective working of the breast unit. Specific economic analysis evaluating main diagnostic services (radiology and pathology) and treatment are needed. The present study assesses the activity level at which the breast unit represents good value for money in surgically-treated patients. Cost assessment is realised by defining a cost function according to the following assumptions: cost function input is personnel costs and technical equipment and output is the number of newly diagnosed cases of primary breast cancer admitted to the breast care unit each year. The increase from 50 new cancer cases per year to 100 will reduce average costs by almost 50%. Cost reduction is important up to a volume of 200 new cases per year. For economic investment to be justified, it is desirable that intake rises to at least 200 new cases per year. Our result is in-line with the EUSOMA recommendation. PMID- 12651200 TI - Age-specific norms and determinants of anxiety and depression in 731 women with breast cancer recruited through a population-based cancer registry. AB - The aim of this study was to determine population norms and determinants of anxiety and depression in a population-based sample of 731 women with breast cancer (aged 23-60 years) with the Hospital Anxiety and Depression scale (HADS). The prevalence of 'probable' psychological morbidity due to anxiety was 23% and due to depression was 3%. When the women identified as 'possible' cases were included, the respective proportions were 45 and 12%. Higher anxiety was present in younger, less educated women not born in Australia. There was no clear pattern of risk factors for depression. These population-based findings highlight the need for clinicians to be aware that age, education and country of birth may identify a particularly vulnerable subgroup. While brief scales such as the HADS are limited in their ability to accurately predict a clinical diagnosis, high scores identify those who may warrant referral for clinical evaluation. PMID- 12651201 TI - Concurrent sequencing of full-dose CMF chemotherapy and radiation therapy in early breast cancer has no effect on treatment delivery. AB - With the increasing use of breast-conserving therapy plus systemic chemotherapy for the treatment of early breast cancer, the optimal sequencing of radiation therapy and chemotherapy remains controversial. Sequencing of therapy may influence not only treatment delivery, but control rates, complications and cosmesis. The aim of this study was to evaluate whether concurrent sequencing of standard doses of CMF (cyclophosphamide, methotrexate and 5-fluorouracil) and adjuvant radiation therapy for early breast cancer impacted on optimum treatment delivery. As both an intravenous (i.v.) 3-week regimen and classic (standard) CMF were utilised in this study, both types of CMF were compared. The effect of sequencing on complications and treatment delays were also assessed. 116 patients treated with CMF chemotherapy and adjuvant tangent breast radiation were studied. 73 patients were treated prospectively with concurrent therapy and were retrospectively compared with a matched group of 40 patients treated with sequential or sandwich therapy. All patients had stage 1 or 2 cancers. There were no planned dose reductions introduced for either treatment modality. Concurrent sequencing had no impact on the ability to deliver optimum radiation or chemotherapy doses. There was no significant difference in acute Radiation Therapy Oncology Group (RTOG) skin reactions or complications between the two groups. Although small, there was a significant delay (1.32 days (0-15 versus 0.36 (0-7)) in the concurrent group (P=0.03) in the delivery of radiation therapy. Sequencing had no significant effect on haematological parameters. 'Standard' CMF had a more profound effect on treatment delivery than i.v. CMF (Radiation delay 2.2 days versus 0.26, P=0.002, % chemotherapy delivered 93% versus 99% P=0.000004). At a mean follow-up of 2.6 years, there was no difference in the cosmetic scores between the two groups. Both local and distant control rates were excellent. This study has shown that standard radiation therapy can be delivered safely concurrently with CMF chemotherapy. Whether this approach may lead to better control rates in the future needs further study. PMID- 12651202 TI - Measurement of an apoptotic product in the sera of breast cancer patients. AB - During apoptosis, a number of intracellular proteins are cleaved by caspases. The intermediate filament protein cytokeratin 18 (CK18) is cleaved at Asp238 and Asp396. A monoclonal antibody, M30, specifically recognises a fragment of CK18 cleaved at Asp396 (M30-antigen). We used an enzyme-linked immunosorbent assay (ELISA) to measure M30-antigen levels in the sera of 82 healthy subjects and 201 patients with breast cancer. Patients with primary cancer had higher M30-antigen levels than healthy subjects (P=0.0001). Patients with recurrent cancer showed higher M30-antigen levels than healthy controls and patients with primary cancer (P<0.0001 and P=0.008, respectively). In patients with primary cancer, M30 antigen levels were higher in the oestrogen receptor (ER)-negative subgroup than the ER-positive subgroup. In patients with recurrent cancer, M30-antigen levels correlated with the number of involved organs and performance status (P=0.041 and P=0.014, respectively). There was no association between serum M30-antigen levels and patient prognosis. We conclude that the levels of circulating M30-antigen are increased in patients with breast cancer. PMID- 12651204 TI - Outcome analysis of 189 consecutive cancer patients referred to the intensive care unit as emergencies during a 2-year period. AB - The referral of critically ill cancer patients to an intensive care unit (ICU) is a matter of controversial debate. This study was conducted by an interdisciplinary clinical group to evaluate the outcome of ICU treatment in cancer patients according to their characteristics at the time of referral. A retrospective analysis was used to identify relevant subgroups among 189 consecutive cancer patients referred as emergencies to one of four ICUs during a 2-year period. Reasons for ICU referral were pneumonia (29.6%), sepsis (27.0%), fungal infection (11.1%), another infection (9.5%), gastrointestinal emergency (16.9%), treatment-related organ toxicity (6.9%), or other, non-infectious complications (43.9%). Vasopressor support was required in 50.3%, mechanical ventilation in 49.7%, and haemodialysis/-filtration in 26.5% of the patients. Overall, 41.3% died during ICU treatment, 12.2% died after transfer from ICU to a non-ICU ward, and 35.4% were discharged alive. Sepsis, mechanical ventilation, vasopressor support, renal replacement therapy and neutropenia were independent risk factors for fatal outcome, but no single risk factor unequivocally predicted death. All patients with fungal infection who required vasopressor support and either had sepsis (n=13) or needed mechanical ventilation (n=14) died during ICU treatment, while all non-septic patients. who did not require mechanical ventilation, were younger than 74 years of age and had a non-infectious underlying complication (n=29), survived. This analysis may help to early identify relevant subgroups of cancer patients with different prognoses under ICU treatment. A prospective study to confirm the predictive usefulness of this approach is needed. Cancer patients should not be excluded from referral to the intensive care unit in an emergency solely due to their underlying malignant disease or a single unfavourable prognostic factor. PMID- 12651203 TI - Analysis of salvage treatments for germ cell cancer patients who have relapsed after primary high-dose chemotherapy plus autologous stem cell support. AB - The aim of this study was to identify treatment strategies and therapeutic or clinical factors that predict for response to salvage therapy and survival in patients with metastatic 'Indiana advanced' or International Germ-Cell Cancer Collaborative Group (IGCCCG) poor prognosis' germ cell cancer (GCT) failing first line sequential high-dose chemotherapy plus autologous stem cell support (HD-CT). A total of 58 'poor prognosis' patients who had relapsed after HD-CT were identified within two large prospective German first-line HD-CT trials (n=286) performed between March 1993 and March 2001. Salvage treatment consisted of the following: cisplatin-based conventional dose CTx+/-resection (19/58; 33%), non cisplatin based CTx (16/58; 28%) or salvage HD-CT (14/58; 24%)+/-resection; resection (n=3) and/or radiation (n=5) only: 7 patients (12%); no specific therapy: 2 patients. 21 (38%) patients responded favourably (Complete Response (CR)/Partial Response (PR) marker-negative) to salvage therapy. The use of salvage HD-CT (2-year survival 48%; P=0.03, the complete resection of residual masses (2-year survival 42%; P=0.015) as well as a favourable response to salvage therapy (2-year survival: 31%, P=0.014) were the only variables on univariate analysis associated with an improved survival. The estimated 2-year overall survival rate is 32% (95% Confidence Interval CI: 29-45%). Approximately 30% of patients relapsing after first-line HD-CT will survive>2 years, particularly those patients who can be treated with a second HD-CT +and/or surgical resection. If feasible, complete surgical resection of residual tumours appears to be the most efficient treatment. PMID- 12651205 TI - Lack of c-kit exon 11 activating mutations in c-KIT/CD117-positive SCLC tumour specimens. AB - Previous studies have shown that STI571, a selective tyrosine kinase inhibitor of c-KIT, is highly effective in c-KIT/CD117-positive gastrointestinal stromal tumours (GIST), especially those that have activating mutations in the c-kit exon 11 that encodes the juxtamembrane (JM) domain of the c-KIT oncoprotein. We examined the prevalence of activating exon 11 c-kit mutations in 26 small-cell lung cancer (SCLC) cases in order to explore whether this disease is also a potential target for treatment with STI571. Expression of c-KIT, estimated by immunohistochemistry, was demonstrated in 14 out of 22 SCLC samples (64%); nine samples showed moderate to strong staining (41%), five samples were weakly positive (23%), whereas eight samples (36%) were negative for CD117. Next, we examined the mutational status of exon 11 of the c-kit gene, by single-stranded conformational polymorphism (SSCP) and sequencing in all of the cKIT/CD117 positive tumours. However, no activating mutations in the c-kit exon 11 were found by either technique. Apparently, c-KIT oncoprotein expression in SCLC was not correlated with activating mutations in c-kit exon 11. In analogy to GISTs, our results could imply that SCLC patients would not benefit from treatment with STI571. PMID- 12651206 TI - Antileukaemic activity of treosulfan in xenografted human acute lymphoblastic leukaemias (ALL). AB - Treosulfan (L-threitol-1,4-bis-methanesulphonate; Ovastat(R)) is a bifunctional alkylating drug indicated for the treatment of advanced ovarian carcinoma. Recent data revealed immunosuppressive characteristics and substantial haematopoietic stem cell toxicity after repeated dosing of mice. Therefore, treosulfan is considered to be an alternative conditioning agent to busulfan (for example) administered prior to allogeneic/autologous stem cell transplantation of patients with haematological malignancies. An antineoplastic activity for treosulfan has been previously shown in preclinical models of melanoma, breast, lung and renal cell carcinomas. Here, in vivo antileukaemic activity of treosulfan is compared with the activity of equitoxic doses of cyclophosphamide or busulfan for the first time using human acute lymphoblastic leukaemia (ALL)-models of paediatric origin xenotransplanted into non-obese diabetic (NOD)/severe combined immunodeficient (SCID) mice. Treosulfan treatment achieved an optimum treated to control (T/C) value of 159% (survival time) against B-ALL-SCID 7 and a T/C value of 0% (tumour growth) against T-ALL-SCID 4 and proB-ALL-SCID 19, respectively. Complete regression of established subcutaneously (s.c.) growing nodules of ALL SCID 4 and 19 was obvious and long-term survivors without tumour re-growth were observed. Equitoxic doses of busulfan (ALL-SCID 4, 7, 19) or cyclophosphamide (ALL-SCID 19) were less effective with regard to the numbers of complete regressions and the number of cured animals. Side-effects included myelotoxicity and a small reduction in body weight, but these were tolerable. Treosulfan can be considered a highly active antileukaemic drug whose corresponding clinical value is to be tested in appropriate protocols with leukaemic patients. PMID- 12651207 TI - Second malignant neoplasms after treatment of childhood cancer. AB - The aim of this study was to determine therapy-related risk factors for the development of second malignant neoplasm (SMN) after childhood cancer. The German Childhood Cancer Registry (GCCR) registers all childhood malignancies since 1980 including SMN. A nested case-control study with 238 SMN cases and 450 controls was conducted. A confirmatory, as well as an explorative, analysis was performed. Radiotherapy showed a small effect on the risk of SMN for doses >or=65 Gy. Regarding the chemotherapeutical agents, we saw increased Odds Ratios (OR) for high doses of cyclophosphamide (CP >8000 mg/m(2) OR=6.3 (95% Confidence Interval (CI): 1.3-30.2)), cisplatinum (DDP >435 mg/m(2) OR=2.8 (95% CI: 1.1-6.7)) and mercaptopurine (MP >5000 mg/m(2) OR=4.5 (95% CI: 1.1-18.9)). Patients jointly receiving high doses of MP (>5000 mg/m(2)) and dexamethasone (DEXA >or=1200 mg/m(2)) had an OR=6.9 (95% CI: 1.2-40.3). Our results could be added to those of other investigations to give indications for modifying future therapeutic strategies for childhood cancer. PMID- 12651208 TI - Unexpected reduction of mortality rates from melanoma in males living in central Italy. AB - A registry-based study has been carried out in central Italy to investigate cutaneous melanoma incidence and mortality trends. The incidence of invasive (1492 cases analysed) and in situ (224 cases) cutaneous melanomas increased significantly from 1985 to 1997, in both genders. The increase of invasive tumours was mainly due to 'thin' ( 0.05). CONCLUSION: Rapid proliferation of rat HSCs occurs in response to AngII treatment, but is inhibited after AT1a receptor is blocked with the antagonist losartan. PMID- 12651234 TI - A new internal fixation system using vertebral pedicle screw for unstable lumbar spine: an in vitro biomechanical stability evaluation. AB - OBJECTIVE: To evaluate the effect of a new internal fixation system utilizing vertebral pedicle screw in fixing unstable lumbar spine in vitro. METHODS: Twelve lumbar spine specimens (L(2 to 5)) obtained from fresh human cadavers were randomly divided into 2 equal groups, namely groups A and B. All intact specimens were subjected to nondestructive testing in 6 loading modalities on a universal testing machine, before complete L(2 to 5) instability was produced by means of a wedge fracture induced by resection of the posterior ligamentous structures. Specimens in group A were fixed with the new internal fixation system, and group B with SOCON pedicle screw. The same nondestructive testing was repeated to determine the structural stability of the spine after fixation in both groups. RESULTS: The injured spines fixed with implants in both groups were significantly more stable than the intact specimens in all modes of tests (P < 0.05), especially in flexion and extension tests, in which a stability increase by 62.8% for flexion and 63.9% for extension was recorded in group A, and 54.9% for flexion and 51.0% for extension in group B. The two fixation systems exhibited similar effect in stabilizing the injured lumbar in all the 6 loading modalities (P < 0.05). CONCLUSION: This internal fixation system with vertebral pedicle screw is capable of providing biomechanical stability for unstable lumbar spine. PMID- 12651235 TI - Expressions of latent membrane protein 1, p53 and bcl-2 proteins and their significance in Hodgkin's lymphoma. AB - OBJECTIVE: To study the expressions of latent membrane protein 1 (LMP1), p53 and bcl-2 proteins and investigate their significance in the pathogenesis of Hodgkin's lymphoma. METHODS: Immunohistochemical staining was used to examine the expressions of LMP1, bcl-2, and p53 proteins in 31 paraffin-embedded tissue samples from Hodgkin's lymphoma. RESULT: The positivity rates of LMP1, p53 and bcl-2 proteins were 58.1% (18/31), 61.3% (19/31) and 35.5% (11/31), respectively, showing a statistically significant difference between the expressions of the former 2 proteins. CONCLUSION: p53 may be involved as an important agent in the pathogenesis of Hodgkin's lymphoma induced by LMP1, whereas bcl-2 appears unrelated to the development of this disease. PMID- 12651236 TI - Immunohistochemical analysis of Fas protein expression in hypertrophic scar and keloid. AB - OBJECTIVE: To find an efficient diagnostic method of hypertrophic scar and keloid. METHODS: Immunohistochemistry was employed to detect the expression and distribution of Fas protein in keloids, hypertrophic scars and their surrounding normal skins. Image analysis was performed to quantitatively compare the differences of the protein expression in these tissues. RESULTS: The expression level of Fas antigen was much higher in the keloids and their surrounding normal skins than in hypertrophic scars, and no significant difference in the expression level between the former 2 tissues was found. In hypertrophic scars, however, much lower Fas protein expression was detected in comparison with that in their surrounding normal skins(P < 0.01). CONCLUSION: Immunohistochemical detection of Fas protein is a simple method that is well applicable in the differential diagnosis of pathological scars. PMID- 12651237 TI - Expression and purification of invasion plasmid antigen C of Shigella flexneri. AB - OBJECTIVE: To induce the expression of and purify invasion plasmid antigen C (IpaC) of Shigella flexneri for studying the pathogenesis of Shigella flexneri. METHODS: Prokaryotic expression plasmid pET32a-ipaC was constructed and incorporated into E.coli BL21 (lambda DE3). The engineered bacteria were induced by isopropyl-beta-D-thiogalactopyranoside (IPTG) to express IpaC, which was identified by SDS-PAGE and purified by QIA expressionist system. RESULTS: SDS PAGE presented a band for the fusion protein with the relative molecular mass of approximately 63 000, whose expression reached up to 11% of the total protein of E.coli BL21(lambda DE3). After proper purification, a purity of the target fusion protein of over 90% was achieved when the concentration of imidazole for elution was 350 mmol/L. CONCLUSION: The recombinant plasmid pET32a-ipaC has been stably and efficiently expressed in E.coli BL21 (lambda DE3), and QIA expressionist purification system proves to be simple and highly efficient. PMID- 12651238 TI - Effects of hyperthermia on rat hippocampal pyramidal cell apoptosis in vitro. AB - OBJECTIVE: To investigate the changes in the morphology and biochemistry of neonatal rat hippocampal pyramidal cells (HPCs)apoptosis in high-temperature environment. METHODS: In vitro culture of HPCs was established and then subjected to environmental stimulation with high temperatures (37, 40, and 42 degrees Celsius respectively) for 1 h. All the cells were collected at different time points after the treatment and examined by flow cytometery, fluorescence staining, agarose gel electrophoresis and in situ cell apoptosis detection kit. RESULTS: The apoptotic rate and the number of apoptotic bodies increased as the temperature was elevated. DNA ladder was presented on agarose gel electrophoresis, and the germination of the apoptotic body was observed by fluorescence microscope. CONCLUSIONS: Hyperthermia may induce the apoptosis in HPCs in vitro, and the extend of apoptosis and apoptotic behavior may vary with the temperature and treatment duration. PMID- 12651240 TI - Effect of Angelica sinensis (Oliv.) on melanocytic proliferation, melanin synthesis and tyrosinase activity in vitro. AB - OBJECTIVE: To investigate the effects of Angelica sinensis (Oliv.) on melanocytes and tyrosinase activity. METHODS: MTT method and cleavage by NaOH were employed to measure the proliferation and melanin synthesis of melanocytes, respectively. Tyrasinase activity assessment was performed by measuring the rate of oxidation of DL-dopa. RESULTS: Angelica sinensis (Oliv.) was found to promote melanocytic proliferation (P < 0.05), resulting in a statistically significant increase in the cell counts (P < 0.05). It also enhanced melanin synthesis (P < 0.05) and tryosinase activity (P < 0.05) of the melanocytes, and the most potent effects were achieved at the concentration of 1 g/L (P < 0.05). CONCLUSION: Angelica sinensis (Oliv.) promotes melanocytic proliferation, melanin synthesis and tyrosinase activity, which may be the mechanism for validating its clinical use in the treatment of skin pigmentation. PMID- 12651239 TI - Molecular cloning and characterization of mannan binding lectin C gene from Balb/c mice. AB - OBJECTIVE: To obtain full-length cDNA encoding mouse mannan-binding lectin C(MBL C) polypeptide. METHODS: The cDNA encoding mouse MBL-C was isolated from Balb/c mouse liver cells by reverse transcriptase (RT)-PCR and inserted into pUC-T vector, and the encoding region structure of the DNA was analyzed to understand its evolutionary relationship with single human MBL homologues. RESULTS: The amplified mouse MBL-C cDNA, which was 735 bp in length, encoded 245 amino acid residues, and its structural analysis showed 100% homology with published mouse MBL-C gene sequence and 71.4% homology with MBL gene of Chinese human. CONCLUSION: The gene encoding Balb/c mouse MBL-C has been successfully isolated, which may facilitate further study of the innate immune functions of MBL molecule in vivo. PMID- 12651241 TI - Enhancement of antitumor effect of cisplatin against human ovarian carcinoma cells by mifepristone in vivo. AB - OBJECTIVE: To investigate in vivo the enhancement by mifepristone (RU486) of the antitumor effect of cisplatin (DDP) on human ovarian carcinoma. METHODS: mouse models bearing xenografted cisplatin-resistant ovarian carcinoma were established, in which the changes in tumor volume and the morphology of the implanted tumors were compared before and after treatment with RU486 and DDP. The expression of glucosylceramide synthase (GcS) mRNA was detected by reverse transcriptase-PCR. RESULTS: The morphology of the tumors in RU486 plus DDP group was significantly different as compared with that of the control group. In comparison with exclusive DDP treatment, combined treatment with RU486 and DDP resulted in significantly greater inhibition rates of the tumors (70.48% vs 21.55%, P < 0.01). GcS mRNA expression was lowered in both RU486 plus DDP and RU486 groups, especially in the former group, as compared with that in control group. CONCLUSION: At a non-toxic dose, RU486 may enhance the sensitivity of tumor cells to cisplatin in vivo, possibly through the mechanism of inhibiting GcS expression at the mRNA level. PMID- 12651242 TI - Preparation of polylactic acid/chitin composite material and its safety evaluation by animal experiments. AB - OBJECTIVE: To prepare a scaffold material with good biocompatibility and biodegradability by compounding polylactic acid (PLA) and chitin. METHODS: After preparation of PLA from lactic acid, the compounding of PLA and chitin was carried out by dissolving these 2 materials in one solution for reaction. The composite material was obtained and molded after the solvent was evaporized, and the safety tests of this resultant material were conducted in guinea pigs and New Zealand rabbits, respectively. RESULTS: In allergic test, the guinea pigs responded to the digestion solution of the composite material in almost the same manner as to normal saline (the latter serving as negative control), and no obvious allergic reaction was observed in the animals except those in positive control group. Pyrogenic test by injecting the digestion solution of the composite material in 6 rabbits found a raise in the body temperature less than 0.2 degrees Celsius, with the total increase (adding up the individual temperature raise) less than 1.0 degrees Celsius, to meet the accepted criteria for the pyrogenic test. In subsequent toxicity test, the rabbits showed no signs of agitation or inanimate behavior after injection. CONCLUSIONS: PLA/chitin composite material conforms to the ISO10993-1, and can be used as a basic scaffold material in tissue engineering. PMID- 12651243 TI - A genetic algorithm for detecting target geometric figures. AB - A genetic algorithm is proposed to detect target geometric figures in an given image. Float-point encoding was adopted to process the parameters of a geometric figures to be detected. On the basis of classical Hough transform, a fitness function was obtained for each individual task, and the individuals with the highest fitness function were identified and copied into the cohort of the next generation. For the rest of the individuals, operation with multi-point crossover or uniform mutation was performed to form new individuals in the next generation. When the termination conditions for this genetic algorithm were met, the best fitted individual was decoded and output as the parameters of the detected geometric figures. This algorithm can eliminate noise interference with good convergence and accurate results, and may save time and storage space during relatively easily programmed computation in comparison with classical Hough transform. PMID- 12651244 TI - In vitro culture of human bone marrow mesenchymal stem cell clonies and induced differentiation into neuron-like cells. AB - OBJECTIVE: To study the long-term in vitro culture of human bone marrow mesenchymal stem cells (hMSC) and their phenotypical and functional properties. METHODS: Adherent hMSC colonies were digested by 0.25% trypsin-EDTA with a clone cycle for in vitro subculture. Flow cytometry was employed to examine the phenotypes of the cells. Their committed differentiation potential to neurons, clone-forming ability and growth curves were all investigated. RESULTS: hMSCs could be subcultured under this culture condition for 20 passages, expressing CD13, CD29 and CD59 but not CD11, CD14, CD31, CD34, CD45, CD80, CD86, CD117 and HLA-DR. The cells could be induced to differentiate into neurons when subcultured for 17 passages. CONCLUSION: hMSCs can be efficiently expanded under this culture condition, and the colony-derived hMSCs can maintain the differentiation potentials and retain their biological characteristics. PMID- 12651245 TI - Anterior decompression and instrumentation with cervical butterfly-like locking plate system for herniated cervical intervertebral discs. AB - OBJECTIVE: To evaluate the clinical efficacy of anterior decompression and internal fixation with cervical butterfly-like locking plate system (CBLP, made in China) in the treatment of cervical intervertebral disc herniation. METHODS: This study included 16 patients with cervical intervertebral disc herniation, aged from 26 to 46 years (36.2 years on average), with the disc hernia occurring in single segment in 12 and 2 segments in 4 cases. The patients received anterior decompression, autogenous iliac bone grafting and internal fixation with CBLP, and follow-up studies lasting for 12 to 24 months (mean 18 months) were conducted in all the cases. RESULTS: All the patients had significant improvements in terms of symptom alleviation without any complications during the follow-up period. Solid bone fusion was achieved within an average period of 3 months, and no screw loosening or plate failure occurred. CONCLUSION: Anterior decompression and internal fixation with CBLP proves to be effective for managing cervical intervertebral disc herniation. PMID- 12651246 TI - Event-related potentials P300 in type 2 diabetes mellitus. AB - OBJECTIVE: To investigate event-related potentials (ERP) P(300) in patients with type 2 diabetes mellitus (DM) for early detection of the abnormalities in cognitive function and studying the related factors. METHODS: With the assistance of Nicolet Vinking IV equipment, ERP (P(300)) were tested in 30 type 2 diabetic patients and 30 healthy subjects matched for age, gender, and education background, and the the results were analyzed statistically. The relationships of each ERP parameter with glucose hemoglobin (GHb) and blood lipid levels and renal function were analyzed. RESULTS: No significant difference was found in P(300) latencies in the control group irrespective of the gender and education (P > 0.05), while the latencies were significantly prolonged (P < 0.01) and the amplitudes decreased (P < 0.05) in diabetic patients. It was found that P(300) correlated with glucose hemoglobin (GHb) and blood lipid levels (P < 0.05). CONCLUSION: ERP test is useful for early detection of cognition dysfunction in DM cases, helping to timely identify diabetic patients with potential dementia. PMID- 12651247 TI - Evaluation of three-dimensional conformal radiation therapy for small primary hepatocellular carcinoma. AB - OBJECTIVE: To evaluate the therapeutic effect of three-dimensional conformal radiation therapy (3D-CRT) on small hepatocellular carcinoma. METHODS: Between May 1998 and Dec 2002, 32 patients with small primary hepatocellular carcinoma underwent 3D-CRT (mean total dose of 55 Gy, ranging from 48 to 63 Gy, given in 6 to 9 fractions completed within 12 to 18 d). The clinical target volumes (CTVs) were covered with 100% isodose curve, using 6 MV X-ray. CT studies before and during a follow-up period of 3 years after 3D-CRT were conducted to evaluate the therapeutic effects, with alpha-feto-protein (AFP) levels also tested. RESULTS: Completed remission of the tumor occurred in 87.5% (28/32) of the cases, and partial remission (PR) was obtained in 12.5% (4/32) of the cases after one year. The 1-year survival rate at stage III was 100%, and 2- and 3-year survival rates were both 97%, with death occurring only in one case because of cerebral hemorrhage. AFP levels of these patients with hepatocellular carcinoma were all significantly decreased (P=0.025). CONCLUSION: 3D-CRT is an effective method in the treatment of patients with small primary hepatocellular carcinoma. PMID- 12651248 TI - Comparative study of serum lipids and serum lipoprotein spectrum in patients with cerebrovascular diseases. AB - OBJECTIVE: To investigate the differences of serum lipids and serum lipoprotein spectrum between patients with cerebral hemorrhage (CH) and cerebral infarction (CI), and study their features of expression in the 2 cerebral conditions. METHODS: Serum levels of triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDLC), very-low-density lipoprotein cholesterol (vLDLC), low-density lipoprotein cholesterol (LDLC) and serum lipoprotein electrophoretogram were determined in 59 CI and 21 CH patients as well as in 30 normal subjects. RESULTS: CI patients had higher levels of TC, LDLC, alpha lipoprotein (alpha LP), beta-lipoprotein (beta LP) and intermediate-density beta lipoprotein (int beta LP) than the normal subjects, and the CH patients had lower LDLC level than the CI patients. The CH patients had also higher levels of beta LP, int beta LP and LDLC than the normal subjects. Progressive regression analysis was performed and resulted in the selection of beta LP, int beta LP and HDLC as the best discriminators for cerebral vascular diseases, which were valid in classifying 71.3% of the cases included in this study. CONCLUSION: beta LP, int beta LP and HDLC are important clinical indicators to predict cerebral vascular diseases. PMID- 12651250 TI - Observation of the short-term therapeutic effect of 3D conformal hypofractionated single high-dose radiotherapy on lung tumors. AB - OBJECTIVE: To evaluate the short-term effects of 3D conformal hypofractionated single high-dose radiotherapy for lung tumors. METHODS: Thirty-four patients with lung tumors that defied surgical removal were treated with 3D conformal hypofractionated single high-dose radiotherapy, and the short-term therapeutic effects and survival rates of the patients were assesses. RESULTS: In 31 (91.2%) of the patients, the local tumor growth was successfully controlled, with the 1-, 2-, and 3- year survival rates of 88.2% (30/34), 35.3% (12/34), and 26.5% (9/34), respectively. CONCLUSION: 3D conformal hypofractionated single high-dose radiotherapy is effective and feasible approach for unresectable lung tumors judging from the short-term effects, but its long-term efficacy awaits further study. PMID- 12651249 TI - Deduction of regional cerebral blood flow loss index after stenosis in cerebral artery. AB - OBJECTIVE: To study the calculation of cerebral blood flow loss ratio after cerebral artery stenosis using the data obtained from transcranial Doppler sonography (TCD), by deducing regional cerebral blood flow loss index (rCBFLI). METHODS AND RESULTS: The data of TCD performed in 31 cases of unilateral stenosis in the middle cerebral artery (MCA) were collected and analyzed for deducing the formula for calculating the MCA stenosis index (STI): STI=1 (Vm(0)/Vm(1)) x (PI(1)/PI(0)), where Vm(0) is the mean normal velocity of MCA blood flow (derived from the measurements in 908 normal subjects at varied ages receiving TCD) and Vm(1) the mean velocity at the stenosed MCA, and PI(0) and PI(1) stand for the pulsatility index before and after stenosis. The ratio of PI(1) to PI(0) is likely to be equivalent to the ratio of Q(1) to Q(0) (Q(0) and Q(1) represent the volume of blood flow in the MCA in normal condition and after stenosis, respectively), thus the equivalence can be extended as rCBFLI=(1 Q(1)/Q(0)) x 100%=(1 PI(1)/PI(0)) x 100%. rCBFLI after MCA stenosis was calculated in 31 cases accordingly, and by means of correlation analysis, we found positive correlation between rCBFLI and Vm(1) (r=0.76, P < 0.001) and between Vm(1) and STI (r=0.85, P < 0.001). In the same way, rCBFLI of 55 sides with MCA stenosis in 43 cases were analyzed, and positive correlation between rCBFLI and Vm (r=0.76, P < 0.001) and between rCBFLI and STI (r=0.83, P < 0.001) was found. CONCLUSION: rCBFLI can be used to evaluate the degree of regional cerebral blood flow loss due to artery stenosis, and the combination of rCBFLI and STI may provide an insight into the changes of cerebral hemodynamics in this pathologic condition. PMID- 12651251 TI - Extracorporeal shock wave lithotripsy for difficulty in withdrawal of double J ureteral stents due to ureteral lithiasis. AB - OBJECTIVE: To explore the efficacy of extracorporeal shock wave lithotripsy (ESWL) to coping with lithiasis-induced difficulty in withdrawing double J ureteral stents. METHODS: This study includes 6 patients with difficulty in withdrawing the stents due to lithiasis, who were in the anterior prone position or posterior oblique supine position to receive ESWL treatment. The stones were localized by double-band intersection X-ray system. The treatment voltage of shock wave was 11.2 kV initially, which was gradually increased to 18.1 kV, and the shock wave attack amounted to 1,000 to 3,000 times in total. RESULTS AND CONCLUSION: After ESWL treatment, the encrusted double J stents were successfully withdrawn in all the 6 cases, without causing any morbidity related to the use of ESWL. Among the various techniques available for removing encrusted double J stents, ESWL appears to be the most effective and the least invasive modality. PMID- 12651252 TI - Synthesis of p, p'-divinylazobenzene. AB - OBJECTIVE: To synthesize p, p'-divinylazobenzene (DVAB). METHOD: Wittig reagent was initially synthesized using P-nitrobenzyl bromide and triphenylphosphine, followed by reaction with formaldehyde to produce p-nitro styrene, which was then deoxidized by Zn/NaOH for the final product of DVAB. RESULTS: Fourier transform infrared spectroscopy, nuclear magnetic resonance and element analysis were employed to identify the structure of DVAB, and the lambda(max) as determined by ultraviolet spectroscopy occurred at 357 nm. CONCLUSIONS: The synthesis procedure of DVAB was therefore optimized with high yield. PMID- 12651253 TI - Interventional therapy for stenosis and aneurysm in renal graft artery: report of 9 cases. AB - OBJECTIVE: To study the interventional therapy for renal graft artery stenosis and aneurysm patients with renal transplantation to further improve the survival rate of the graft. METHOD: Seven patients with of renal graft artery stenosis received balloon dilatation of the stenotic artery, followed by stent implantation. For renal graft artery aneurysm in another 2 patients, thrombin infusion and stent implantation were respectively performed. RESULTS: The condition was successfully managed in 6 of the 7 patients with renal artery stenosis, whose serum Cr levels dropped to below 106 micromol/L 3 d after the operation. Thrombin infusion in one of the 2 patients with renal artery aneurysm caused thrombus in the renal graft and then aneurysm rupture, resulting in final graft loss. The other aneurysm case was successfully managed with stent implantation. CONCLUSIONS: Interventional therapy as balloon dilatation combined with stent implantation is ideal for treating renal graft artery stenosis, and stent implantation constitutes an important management for artery aneurysm in the renal graft. PMID- 12651254 TI - Clinical effect of removable partial denture prepared by Saddle-Lock hidden clasp. AB - OBJECTIVE: To study the method for preparing removable partial denture using Saddle-Lock hidden-clasp and evaluate its clinical effect. METHOD: Casting process was adopted for the preparation of the clasp, which was fixed at the points on the mesial and distal surfaces of the abutment tooth. The clinical records of 305 patients treated with such removable partial dentures were reviewed. RESULTS: Within a follow-up period of 6 years, 295 of the 305 patients retained good abutment condition and were satisfied with the denture that was capable of easy wearing and removing. CONCLUSIONS: Removable partial denture made by Saddle-lock hidden-clasp exhibits good performance in fixation and stability. As an innovation of traditional design of partial denture clasp, it is simple to prepare and well received by the patients with good clinical effect. PMID- 12651255 TI - Unrelated bone marrow transplantation for beta-thalassemia major: report of the first 2 successful cases in Asia. AB - We report our experience in successful unrelated-donor bone marrow transplantation in 2 cases of beta-thalassemia major, which are approved to be the first 2 cases in Asia. The 2 children receiving transplantation of the bone marrow from unrelated donors were diagnosed as having beta-thalassemia major, whose gene mutation type was homozygous and double heterozygote. High-resolution HLA typing found a mismatch with 1 sub-locus in both cases, and red blood cell type was mismatched in 1 case. Pretransplant conditioning protocol contained busulfan (BU, 16 mg/kg x b.w.), cyclosphamide (CY, 200 mg/kg x b.w.), anti thymocyte globulin (ATG, 90 mg/kg x b.w.) and fludarabine (25 mg x d-1 x m-2). To prevent graft-versus-host disease (GVHD) episodes, cyclosporine-A (Cs-A) and methotrexate (MTX) were administered. The 2 cases experienced condition resembling serum sickness during pretransplant conditioning. After bone marrow transplantation, 1 case had grade acute GVHD in the skin, another developed grade acute GVHD in both the skin and intestinal tract, but the condition was brought under control in both cases after proper treatment. The time for peripheral neutrophil granulocyte recovery to above 0.5 x 10(9)/L was 19 and 16 days postoperatively, respectively, and the time of WBC recovery to normal was 54 and 80 days postoperatively. Platelet recovery to over 50 x 10(9)/L occurred on postoperative days 61 and 90, and Hb recovered to above 100 g/L in both case on days 110 and 83 respectively. The time of final blood transfusion was 53 and 62 days postoperatively for the 2 patients. Gene mutation type in the 2 cases was switched to normal of the donors. After retrieval of relative literature, the 2 cases were approved as the first 2 successful unrelated-donor bone marrow transplantation cases in Asia. This will provide a new possibility of donor supply for hematopoietic stem cells transplantation in thalassemia. PMID- 12651256 TI - Effect of estrogen and progestogen on nitric oxide content in rat endometrium. AB - OBJECTIVE: To observe the effect of estrogen and progestogen on the content of nitric oxide (NO) in the endometrium of SD rats. METHOD: Fifteen SD rats were divided into control, estrogen and levonorgestrel groups (5 in each group), with corresponding treatment indicated by the designation of the groups. The NO contents in the endometrium were determined by Griess method. RESULTS: he NO content of estrogen group was significantly higher, while that of levonorgestrel group significantly lower, than that of the control group (P < 0.01 for both comparisons). CONCLUSIONS: Estrogen and levonorgestrel may regulate NO synthesis in the endometrium. PMID- 12651257 TI - Current use of imatinib in the treatment of chronic myeloid leukemia. PMID- 12651258 TI - Intense immunosuppression and autologous hematopoietic stem cell transplantation for multiple sclerosis. PMID- 12651259 TI - Chronic graft versus host disease after allogeneic peripheral blood transplantation. PMID- 12651260 TI - Advances in unrelated donor hematopoietic cell transplantation. PMID- 12651261 TI - Prevalence of C282Y and E168X HFE mutations in an Italian population of Northern European ancestry. AB - BACKGROUND AND OBJECTIVES: In Italy, the prevalence of C282Y is lower than in Northern European countries. We hypothesized a higher prevalence of C282Y in Northern than in Southern Italian populations. We previously identified a nonsense mutation (E168X) in hemochromatosis probands originating from a region in the north-west of Italy. We aimed to define the prevalence of C282Y and E168X in that region and the origin of the E168X mutation by haplotype analysis. DESIGN AND METHODS: Six-hundred and six blood donors were investigated for C282Y, H63D, S65C and E168X mutations by polymerase chain reaction (PCR)-restriction assays. Three hundred were also tested for rare HFE and TFR2 mutations by reverse hybridization test strips. D6S265, D6S105 and D6S1281 microsatellites were analyzed to define E168X 6p-associated haplotypes. RESULTS: One C282Y homozygote, thirteen C282Y/ H63D compound heterozygotes, four E168X heterozygotes and three E168X/H63D compound heterozygotes were found. The allele frequencies of C282Y, H63D, S65C, and E168X were 4.7%, 14.9%, 0.74% and 0.58%, respectively. INTERPRETATION AND CONCLUSIONS: The prevalence of C282Y in the region investigated was much higher than that previously reported in Italy. This finding is probably due to the heavy Celtic component of this north-western population and suggests that in populations of Northern Italian descent screening studies for hemochromatosis could be cost-effective. The prevalence of E168X in this region, although low, suggests that the mutation probably originated here many years ago and its frequency increased as a result of a local founder effect. Given its severity, we suggest that the E168X mutation should be searched for in all hemochromatosis patients of Northern ancestry with an incomplete HFE genotype. PMID- 12651262 TI - Risk and early cytogenetic response to imatinib and interferon in chronic myeloid leukemia. AB - BACKGROUND AND OBJECTIVES: We compared the early cytogenetic response (CgR) to a combination of imatinib mesylate (Glivec, Novartis Pharma, Basel, Switzerland) and a pegylated form of human recombinant interferon-alpha2b (pegIFN-alpha2b, PegIntron, Schering Plough, Kenilworth, New Jersey, USA) with the relative risk, either according to Sokal's or Euro scoring systems. DESIGN AND METHODS: Seventy seven patients with early chronic phase, previously untreated, Ph-positive chronic myeloid leukemia (CML) received a combination of imatinib mesylate (400 mg/day) and pegIFN-alpha2b (3 consecutive cohorts treated with 50, 100 or 150 mg/weekly). Fifty-seven patients have completed the first 6 months of treatment and are evaluable for CgR. RESULTS: After 6 months of treatment, the overall major CgR rate was 89% and 90% in low risk patients (Sokal's and Euro, respectively), 76 and 59% in intermediate risk and 23% and 17% in high risk patients. These differences were significant (p=0.0001 for Sokal and 0.001 for e). INTERPRETATION AND CONCLUSIONS: For the first time, these data suggest that the early CgR rate to a imatinib mesylate-based regimen is significantly risk related. PMID- 12651263 TI - Chronic myeloid leukemia in chronic phase responding to imatinib: the occurrence of additional cytogenetic abnormalities predicts disease progression. AB - BACKGROUND AND OBJECTIVES: The acquisition of additional cytogenetic changes (clonal evolution, CE) during treatment of chronic myeloid leukemia (CML) with imatinib mesylate is currently regarded as an index of increasing resistance to imatinib. Therefore, to investigate whether CE as an isolated event increases the risk of disease progression during imatinib treatment, we compared the outcome of patients with CML in chronic phase (CML-CP) who developed CE whilst in complete hematologic remission with the outcome of comparable patients in complete hematologic remission who showed no evidence of CE. DESIGN AND METHODS: We serially studied cytogenetic findings in 102 patients receiving the Abl-tyrosine kinase inhibitor, imatinib mesylate, as sole agent to treat CML-CP who had no evidence of CE before initiation of imatinib treatment. RESULTS: CE was identified during treatment with imatinib in 15 patients, 10 of whom were in complete hematologic remission. In most cases these changes occurred exclusively in the Ph+ population but in three patients additional changes occurred in a co existing Ph-negative population. Patients with de novo CE in the absence of any other sign of disease progression had a significantly higher incidence of progression by 18 months than did non-CE patients (progression-free survival 34.3% (CI 10.5-69.8%) vs. 94.1% (CI 80.6-98.4%), p<0.0001). INTERPRETATION AND CONCLUSIONS: Based on this relatively small series of patients, we conclude that acquisition of clonal evolution increases the risk of subsequent disease progression also in CML patients in complete hematologic remission on imatinib. PMID- 12651265 TI - A multiplex reverse transcriptase-polymerase chain reaction strategy for the diagnostic molecular screening of chimeric genes: a clinical evaluation on 170 patients with acute lymphoblastic leukemia. AB - BACKGROUND AND OBJECTIVES: In the last few years molecular methods have allowed the identification of leukemia-associated genetic lesions, which may represent the most accurate predictors of clinical outcome. These considerations strengthen the need for rapid identification of the abnormalities. Our aim was to demonstrate whether a modified multiplex reverse transcription polymerase chain reaction (RT-PCR) system might be successfully used to screen a large number of patients with acute lymphoblastic leukemia (ALL). DESIGN AND METHODS: In this study we adapted the multiplex RT-PCR assay, previously described by Pallisgaard et al., to detect all the most frequent genetic lesions with their characteristic splicing variants occurring in acute lymphoblastic leukemia, such as the MLL/AF4, MLL/ENL, BCR/ABL p190 (e1a2) and p210 (b2a2,b3a2) isoforms, E2A/PBX1, TEL/AML1, SIL/TAL1 and the novel NUP98/RAP1GDS1 transcript, recently described in a T-ALL leukemic subtype. RESULTS: We used the multiplex RT-PCR assay to screen 170 ALL patients (70 children and 100 adults). PCR positivity was detected in 67 (39%) of the 170 ALL patients studied. The comparison between cytogenetic and molecular analyses showed complete correspondence between the two assays in all patients with an evaluable karyotype. Finally, the observed incidence of genetic lesions in our ALL patients was similar to the frequency usually reported both in children and in adults with ALL. INTERPRETATION AND CONCLUSIONS: These results show that, compared to single RT-PCR reactions, our multiplex RT-PCR system allows rapid, specific, simultaneous as well as a less expensive, laborious and time-consuming detection of the most frequent fusion transcripts in ALL patients. Therefore, it might be recommended for rapid diagnostic molecular screening of large numbers of patients, such as those enrolled in multicenter, co-operative studies. Furthermore, we have shown that multiplex RT-PCR is an open system that can easily be adapted to detect new leukemic genes. PMID- 12651264 TI - Expression of recombination activating genes-1 and-2 immunoglobulin heavy chain gene rearrangements in acute myeloid leukemia: evaluation of biological and clinical significance in a series of 76 uniformly treated patients and review of the literature. AB - BACKGROUND AND OBJECTIVES: Early lymphoid differentiation is characterized by antigen receptor gene rearrangements; the rearrangement process is governed by two lymphoid-specific genes, RAG (recombinase activating gene)-1 and -2. The available data on the incidence and prognostic significance of clonal immunoglobulin heavy chain (IgH) gene rearrangements in acute myeloid leukemia (AML) are rather contradictory. The aim of this study was to evaluate the incidence and prognostic significance of RAG-1 and -2 mRNA transcripts and clonal IgH gene rearrangements in a cohort of uniformly treated AML patients; the available literature is also reviewed. DESIGN AND METHODS: The study was performed on 76 AML patients, newly diagnosed between August 1996 and November 1999. RAG-1/-2 gene expression was analyzed by a reverse transcriptase polymerase chain reaction technique and IgH gene rearrangements were detected with variable region (VH) family-specific and consensus framework region (FWR)-2 and/or-3 primers. Statistical associations were explored between IgH monoclonality/ RAG mRNA expression and: (i) age, gender, FAB subtype, immunophenotype, cytogenetic risk groups; (ii) response variables (response/relapse incidence, survival). RESULTS: In total, 38/75 samples (50.6%) were RAG-1 and/or -2 positive; 30/76 samples (39.5%) carried clonal IgH genes, whereas 13/30 IgH-positive samples (43.3%) were RAG-1/2-negative. Significant associations were detected only for RAG-2 positivity and unfavorable karyotype and IgH monoclonality and FAB subtypes M4/ M5; no association was identified with response outcome and survival. INTERPRETATION AND CONCLUSIONS: Lymphoid-specific molecular markers are detected in a significant proportion of AML patients, regardless of differentiation status (assessed morphologically/ immunophenotypically); however, in our experience, they do not seem to constitute an adverse prognostic factor. PMID- 12651266 TI - Primary bone lymphoma: experience with 52 patients. AB - BACKGROUND AND OBJECTIVES: A retrospective analysis was performed to assess the efficacy of various treatments of non-Hodgkin's primary bone lymphomas (PBL). DESIGN AND METHODS: Fifty-two consecutive, previously untreated PBL patients were seen between the years 1982 and 1998. Information was obtained regarding each patient's presentation and clinical course. Histology was reviewed in all cases. Modern immunohistochemical stains were performed on each case. RESULTS: Regarding therapeutic approach, we observed a complete response (CR) in 35/41 (85%) patients treated with chemotherapy with/without radiation therapy and in 7/11 (64%) patients who received radiation therapy alone. Relapses were observed in only 2/35 (6%) patients after chemotherapy (with/without radiation therapy), as compared with 4/7 (57%) patients after radiation therapy alone (p = 0.004); the relapse-free survival curves of these two subsets were significantly different. At both univariate and multivariate analysis only type of front-line therapeutic approach (chemotherapy with/without radiation therapy vs. radiation therapy alone) turned out to have a significant prognostic influence. INTERPRETATION AND CONCLUSIONS: Our data indicate that in PBL use of chemotherapy or combined modality therapy seems to provide more durable CRs than radiation therapy alone. PMID- 12651268 TI - Allogeneic and autologous bone marrow transplantation after consolidation therapy in high-risk acute myeloid leukemia in children. Towards a risk-oriented therapy. AB - BACKGROUND AND OBJECTIVES: Although chemotherapy in childhood acute myeloid leukemia (AML) has improved in the last decade, except for a group of better-risk patients (approximately one third), more than half the other patients relapse. The main objective of this study was to evaluate the results obtained with bone marrow transplants, either allogeneic (allo-BMT) or autologous (auto-BMT), following two intensive consolidation courses in a series of children with high risk (HR) AML according to morphologic and early-response BFM criteria. A second objective was to compare the results of auto-BMT with those of allo-BMT. DESIGN AND METHODS: From April 1988 to May 2001, 79 children (< 15 years old) with de novo AML entered the prospective AML-88 trial in a single institution: 50 (63%) were qualified as having high-risk disease and are the subject of this study. After 1 or 2 induction courses, depending on early response, and two consolidations, patients with an HLA-identical sibling received an allo-BMT and all the others an auto-BMT. The conditioning regimen was cyclophosphamide and total body irradiation (TBI) in children over 3 years old and busulfan and etoposide in younger children. Bone marrow was purged with mafosfamide in auto BMT and cyclosporine alone was given as graft-versus-host disease (GVHD) prophylaxis in allo-BMT. RESULTS: At the end of the chemotherapy phase (induction and consolidation ), 46 of the 50 HR patients (92%) had attained complete remission (CR) after one (n=29), two (n=11) or three (n=6) courses; 2 more were in partial remission (PR) and 2 had died. The 48 patients in CR or PR received either an allo-BMT (17) or an auto-BMT (31). Hematologic reconstitution was significantly slower in auto-BMT recipients. Forty-one percent of patients who received allo-BMT suffered acute GVHD grades II-IV. Toxic deaths and relapse rates were 5.9% and 17.6%, respectively, in allo-BMT and 3.2% and 25.8%, respectively, in auto-BMT. Post-transplant 8-year event-free survival (EFS) was 74.5% (54-96) in allo-BMT and 74.2% (59-89) in auto-BMT. EFS and OS in all the series (50 patients) were 71% (59-83) and 73% (61-85), respectively, with a median follow-up of 7.2 years. INTERPRETATION AND CONCLUSIONS: This study indicates that improved results in children with HR-AML can be obtained by either allo- or auto-BMT performed after two courses of intensive consolidation therapy provided good supportive therapy is given and reduced transplant -related mortality (TRM) is minimized. PMID- 12651267 TI - The factor V (FV) gene ASP79HIS polymorphism modulates FV plasma levels and affects the activated protein C resistance phenotype in presence of the FV Leiden mutation. AB - BACKGROUND AND OBJECTIVES: In carriers of the factor V (FV) Leiden mutation, different trans-acting gene variants (HR2 haplotype and FV Cambridge mutation) affect activated protein C (APC) sensitivity. Among a series of FV gene variants characterized, the Asp79His polymorphism appeared to be a good candidate for the modulation of FV activity. DESIGN AND METHODS: In a group of 150 apparently healthy subjects without the FV Leiden mutation and in 55 apparently healthy subjects with mutation, genotypes of the Asp79His polymorphism and of the HR haplotype were characterized and plasma levels of FV coagulant activity and APC ratios evaluated. RESULTS: In the group without the FV Leiden mutation, 16 subjects (10.7%) carried the His 79 allele and 15 subjects (10.0%) the HR2 haplotype. Two of them carried both gene variants. As compared to FV activity levels in non-carriers (106.4+18.5%), values were lower in subjects with the His79 allele (95.2+25.2%; p=0.025) and in those with the HR2 haplotype (93.7+16.2%; p =0.007). FV activity levels were further reduced in carriers of both FV gene variants (78.7+3.3%; p =0.009). APC values were similar among individuals carrying different FV genotypes. In the group with the FV Leiden mutation, APC ratios were lower in subjects carrying the His 79 allele (0.63; p =0.008) or the HR2 haplotype (0.63; p =0.026) than in subjects without (0.69) reflecting FV activity values. INTERPRETATION AND CONCLUSIONS: Present data suggest that carriership of the His79 allele modulate plasma levels of FV coagulant activity and, in subjects carrying the FV Leiden mutation, affects APC sensitivity. PMID- 12651269 TI - Low transplant related mortality in older patients with hematologic malignancies undergoing autologous stem cell transplantation. AB - BACKGROUND AND OBJECTIVES: Patients over 60 years are frequently excluded from autologous stem cell transplantation (ASCT) programs due to a traditionally high rate of transplant-related mortality (TRM) in such indications. We evaluated the results of ASCT in a group of 49 patients >= 60 years of age [32 males, median age 63 years (range, 60 to 71)] autografted in our institution from January 1995 to December 2001. DESIGN AND METHODS: There were 27 patients with multiple myeloma, 13 with non-Hodgkin's lymphoma, 3 with acute myelogenous leukemia, 3 with chronic myelogenous leukemia and 3 with other hematological malignancies. The Karnofsky score was >= 80% in 47 cases. The median time from diagnosis to ASCT was 12 months (range, 5 to 61). Twenty-four patients were autografted in an early disease phase and 25 (51%) in an advanced phase. Peripheral blood stem cells were used in 46 patients (94%), bone marrow in one (2%) and bone marrow plus peripheral blood in two (4%). Forty-one patients received chemotherapy-only conditioning regimens, while only 8 patients received total body irradiation. RESULTS: Engraftment occurred in all but one patient. The median times to achieve a sustained absolute neutrophil count > 0.5 x 10(9)/L and a sustained platelet count >20 x 10(9)/L were 13 (range, 10 to 35) and 13 days (range, 8 to 62), respectively. The actuarial 2-year overall survival was 67% [95% confidence interval (CI), 52-82%). Four patients died without progression due to central nervous system (CNS) hemorrhage (n = 1), CNS toxicity (n = 1), fungal infection (n = 1) or toxoplasmosis (n = 1). One hundred-day and 1-year actuarial TRM were 4% (95% CI, 1-16%) and 8% (95% CI, 3-21%), respectively. INTERPRETATION AND CONCLUSIONS: ASCT is a feasible procedure in selected elderly patients, with apparently similar rates of engraftment and TRM to those reported for younger patients. PMID- 12651270 TI - CD34+ selected autologous peripheral blood stem cell transplantation for multiple sclerosis: report of toxicity and treatment results at one year of follow-up in 15 patients. AB - BACKGROUND AND OBJECTIVES: Autologous stem cell transplantation (ASCT) is currently being evaluated as a therapy for patients with multiple sclerosis (MS). We report the results of a phase II trial to evaluate feasibility and toxicity of CD34+ selected ASCT (CD34+/ASCT) and treatment results at one year of follow-up. DESIGN AND METHODS: Patients with advanced secondary progressive (SP) or relapsing-remitting (RR) MS and confirmed worsening of the extended disability status scale (EDSS) in the previous year despite interferon or other immunotherapies were included. Peripheral blood stem cells were obtained by leukaphereses after mobilization with cyclophosphamide (Cy) and granulocyte colony-stimulating factor (G-CSF). CD34+ selection was performed by means of an Isolex 300 or CliniMACS device. BCNU, Cy and antithymocyte globulin (ATG) were administered as conditioning regimen. RESULTS: Fifteen patients (9 SPMS and 6 RRMS) with a median EDSS of 6.0 (4.5-6.5) and a median of 3 (1-7) relapses in the previous year were included. Mobilization was unsuccessful in one patient. During mobilization, one patient had a transient neurologic deterioration. The main complication during ASCT were engraftment syndrome, which developed in three patients, CMV reactivation in one, and neurologic deterioration in two patients coinciding with high-fever related to ATG. Hematologic recovery was fast and complete in all cases. At 12 months, the EDSS had improved in three patients, worsened in two and remained stable in nine. Despite withdrawal of all immunosuppressive therapy only two patients had relapses. Magnetic resonance imaging showed disappearance of enhanced T1 lesions but oligoclonal bands persisted in the cerebrospinal fluid of all evaluated cases. INTERPRETATION AND CONCLUSIONS: CD34+/ASCT using BCNU, Cy and ATG as conditioning regimen has an acceptable toxicity and clearly reduces the progression of MS. Further follow-up is necessary to establish the real impact of this procedure on the long-term evolution of the disease. PMID- 12651271 TI - One-year cyclosporine prophylaxis reduces the risk of developing extensive chronic graft-versus-host disease after allogeneic peripheral blood stem cell transplantation. AB - BACKGROUND AND OBJECTIVES: Chronic graft-versus-host disease (GVHD) remains the most common late complication of allogeneic stem cell transplantation, producing significant long-term morbidity and contributing to a substantial risk of late mortality. Chronic GVHD may be more common, more protracted and less responsive to current treatments after peripheral-blood stem cell (PBSC) transplantation than after bone marrow transplantation. The purpose of this retrospective cohort study was to determine whether the hazard of extensive chronic GVHD after allogeneic PBSC transplantation could be decreased by prolonging cyclosporine A (CsA) prophylaxis over 12 months. DESIGN AND METHODS: Fifty-seven consecutive patients with hematologic malignancies who had received a PBSC transplant from an HLA-identical sibling were evaluable for chronic GVHD. All patients began CsA tapering at day 50 but 2 different durations of immunosuppression were used: the first 36 patients were allocated to receive a 6-month course with tapering by 5% at weekly intervals (group A), while the following 21 received a 12-month course with tapering by 5% every 2 weeks (group B). RESULTS: The cumulative incidence of extensive chronic GVHD at 2 years was 69% (95% CI, 53-85%) for group A and 25% (95% CI, 3-47%) for group B with a significantly lower hazard in group B than in group A (HR=0.2; 95% CI, 0.07-0.57; p=0.0009). In multivariate analysis, the 12 month CsA tapering schedule was associated with a significantly decreased risk of extensive chronic GVHD (HR=0.2; 95% CI, 0.06-0.66; p=0.008). The hazard of transplant-related mortality, relapse and failure to survive in remission was not significantly different among the 2 groups. INTERPRETATION AND CONCLUSIONS: One year CsA prophylaxis seems to be more effective than the standard six-month CsA regimen at preventing extensive chronic GVHD after PBSC transplant from an HLA identical sibling. Conclusive assessment of the benefits of such prolonged immunosuppression, in terms of better quality of life and minor morbidity, requires both long-term follow-up to evaluate the rates of relapse and secondary tumors and a randomized setting. PMID- 12651272 TI - Comparison of cytotoxicity of 2-chloro- 2'-arabino-fluoro-2'-deoxyadenosine (clofarabine) with cladribine in mononuclear cells from patients with acute myeloid and chronic lymphocytic leukemia. AB - BACKGROUND AND OBJECTIVES: Clofarabine (CAFdA), one of the newer nucleoside drugs is undergoing a phase II clinical trial for the treatment of pediatric refractory/relapsed acute myeloid and lymphocytic leukemia. Although CAFdA is structurally similar to the clinically established analogs fludarabine and cladribine (CdA), its metabolism and mechanism of actions are significantly different. The present study investigates the in vitro cytotoxicity of CAFdA and CdA in mononuclear cells isolated from 52 patients with chronic lymphocytic (CLL) and acute myeloid leukemia (AML). DESIGN AND METHODS: We incubated the leukemic cells with drugs for 48 hours and cytotoxicity was then evaluated by the MTT dye assay. We also determined the levels of deoxycytidine and deoxyguanosine kinase with radio-chemical substrate-based assays and used a high performance liquid chromatographic method to measure cellular nucleotides in leukemia cells after 2 hours' incubation. RESULTS: Using equimolar concentrations of CAFdA and CdA, the in vitro cytotoxicity for the population was significantly higher with CAFdA than with CdA (median EC50 for CAFdA 0.12 microM and for CdA 0.15 microM, p<0.001). From the individual estimates the difference in cytotoxicity between CAFdA and CdA was more pronounced in cells from CLL patients (median EC50 for CAFdA 0.08 microM and for CdA 0.16 microM p<0.001) than in those from AML patients. We also found that CAFdA was phosphorylated more efficiently than CdA. No correlations were detected in this study between the levels of CdA and CAFdA nucleotides, enzymes levels and the in vitro responses. INTERPRETATION AND CONCLUSIONS: The greater in vitro cytotoxicity and cell metabolism of CAFdA compared to CdA confirm the high activity of CAFdA and encourage clinical trials with CAFdA in leukemic patients. PMID- 12651273 TI - Second-generation blood tests to detect erythropoietin abuse by athletes. AB - BACKGROUND AND OBJECTIVES: We previously developed blood tests that were introduced at the Sydney 2000 Olympic Games to identify athletes injecting recombinant human erythropoietin (rHuEPO). The aim of this study was to re analyse our existing database to develop models with heightened sensitivity, using wherever possible blood parameters measurable with appropriate standards of analytical performance. DESIGN AND METHODS: The principal database for this study was derived from a double-blind trial in which 57 recreational athletes were administered either rHuEPO or placebo. Standard discriminant analysis was used to derive two ON models (ON-hes and ON-he) and two OFF models (OFF-hr and OFF-hre) sensitive to accelerated and decelerated erythropoiesis respectively, utilising concentrations of hemoglobin (h), erythropoietin (e) and serum transferrin receptor (s), as well as percent reticulocytes (r). The ability of our models to detect rHuEPO administration was assessed by comparing model scores of subjects in the administration trial with the model scores of 1152 elite athletes from 12 countries. RESULTS: The ability of the new models to detect rHuEPO administration was generally higher than that of our previous models, particularly during phases when low doses of rHuEPO were used, and after injections had ceased. INTERPRETATION AND CONCLUSIONS. The increased stability of the new blood parameters facilitates transport of samples to central laboratories, and the heightened sensitivity of the new models makes them better than existing models for federations wishing to screen samples for urine testing and to identify and target suspect athletes for out-of-competition testing. However procedures should be incorporated that respect an elevated model score caused by genetic, health or environmental circumstances. PMID- 12651274 TI - Transferrin receptor-2 polymorphisms and iron overload in transfusion independent b-thalassemia intermedia. PMID- 12651275 TI - Molecular characterization of glucose-6-phosphate dehydrogenase deficiency in the Fars province of Iran. PMID- 12651276 TI - Soluble transferrin receptor in iron-deficient patients with and without anemia. PMID- 12651277 TI - Quantitative real-time polymerase chain reaction shows that treatment with interferon reduces the initially upregulated PRV-1 expression in polycythemia vera patients. PMID- 12651278 TI - Acute myeloid leukemia with recurring chromosome abnormalities as defined by the WHO-classification: incidence of subgroups, additional genetic abnormalities, FAB subtypes and age distribution in an unselected series of 1,897 patients with acute myeloid leukemia. PMID- 12651279 TI - Thymidylate synthase gene polymorphism and its association with relapse in childhood B-cell precursor acute lymphoblastic leukemia. PMID- 12651280 TI - Simultaneous expression of CD38 and its ligand CD31 by chronic lymphocytic leukemia B-cells: anecdotal observation or pathogenetic hypothesis for the clinical outcome? PMID- 12651281 TI - Ultrasound-guided fine needle aspiration cytology combined with flow cytometric immunophenotyping for rapid characterization of deep-seated non-Hodgkin's lymphoma recurrence. PMID- 12651282 TI - Low-grade non Hodgkin's lymphomas in the elderly: impact of a low-dose fludarabine-based combination regimen (mini-FLEC). PMID- 12651283 TI - Successful treatment with voriconazole of cerebral aspergillosis in an hematologic patient. PMID- 12651284 TI - Chemotherapy alone may lead to a PCR negative stem cell harvest in transformed lymphoma refractory to rituximab. PMID- 12651285 TI - Red blood cell phosphosphoglycerate mutase. Description of the first human BB isoenzyme mutation. PMID- 12651286 TI - The first case of a compound heterozygosity for Hb E-Saskatoon and HbS. PMID- 12651287 TI - Idiopathic myelofibrosis with fatty bone marrow: an issue of sampling? A propos an unusual case. PMID- 12651288 TI - Treatment of severe life-threatening graft-versus-host disease by autologous peripheral blood stem cell transplantation using a nonmyeloablative preconditioning regimen. PMID- 12651289 TI - Pentanucleotide repeat (TTTTA)n polymorphism in the 5' control region of the apoliprotein (A) gene and atherothrombotic serum lipoprotein (A) concentration, in a pediatric population. PMID- 12651290 TI - The location of deep-vein thrombosis as a predictive factor for recurrence and cancer discovery after proximal deep-vein thrombosis. PMID- 12651291 TI - About mediastinal large B-cell lymphoma and other lymphoma entities. PMID- 12651292 TI - The CD4+ CD56+ CD116- CD123+ CD45RA+ CD45RO- profile is specific of DC2 malignancies. PMID- 12651293 TI - Organic and inorganic sulfur transport in the xylem sap and the sulfur budget of Picea abies trees. AB - Temporal changes in inorganic and organic sulfur compounds (sulfate, glutathione, cysteine, methionine) were analyzed in xylem sap of 40-year-old Norway spruce (Picea abies (L.) Karst.) trees growing on acidic soils at a healthy and a declining stand in the Fichtelgebirge (North Bavaria, Germany). Studies were carried out (1) to quantify glutathione (GSH) transport in the xylem of spruce, (2) to study the significance of reduced sulfur versus sulfate (SO(4) (2-)) transport in the xylem, and (3) to compare total sulfur (S) transport in the xylem with the amount of foliar uptake of SO(2) in an air-polluted environment. Glutathione was the main reduced S compound in the xylem ranging in concentration from 0.5 to 5 &mgr;mol l(-1). Concentrations of inorganic SO(4) (2-) in the xylem sap were up to 50 times higher than those of GSH ranging from 60 to 230 &mgr;mol l(-1). During the growing season, concentrations of all S compounds in the xylem were highest in May (up to 246 &mgr;mol l(-1)) and decreased during summer and fall (up to 21 &mgr;mol l(-1)). On average, SO(4) (2-) concentrations in xylem sap were 30% higher at the declining site compared with the healthy site. Diurnal changes in organic S compounds were significant for GSH and cysteine with high concentrations during the night and low concentrations during the day. Diurnal changes in inorganic concentrations were not significant. Xylem sap concentrations of SO(4) (2-) and cysteine were twice as high and GSH concentrations were tenfold higher in surface roots than in branches. At both sites, transport of organic S was low (up to 3% of total S) compared to transport of SO(4) (2-). Annual transport of total S in the xylem (SO(4) (2-) was the main component) ranged from 60 to 197 mmol tree(-1) year(-1) at the healthy site and from 123 to 239 mmol tree(-1) year(-1) at the declining site. Although gaseous uptake of SO(2) was estimated to be similar at both sites (38 mmol tree(-1) year( 1); Horn et al. 1989), the ratio between annual gaseous uptake of SO(2) and transport of S in the xylem was 1:4 and 1:5 at the healthy and declining sites, respectively. PMID- 12651294 TI - Effects of temperature and tissue nitrogen on dormant season stem and branch maintenance respiration in a young loblolly pine (Pinus taeda) plantation. AB - We measured dormant season (November through February) maintenance respiration rates (R(m)) in stems and branches of 9-year-old loblolly pine (Pinus taeda L.) growing in plots under conditions of controlled nutrient and water supply in an effort to determine the relationships between R(m) and tissue size (surface area, sapwood volume, sapwood dry weight), tissue nitrogen content and temperature. Dormant season R(m) per unit size (i.e., surface area, &mgr;mol m(-2) s(-1); sapwood volume, &mgr;mol m(-3) s(-1); or sapwood dry weight, nmol g(-1) s(-1)) varied with tissue size, but was constant with respect to tissue nitrogen content (&mgr;mol mol(-1) N s(-1)). Cambium temperature accounted for 61 and 77% of the variation in stem and branch respiration, respectively. The basal respiration rate (respiration at 0 degrees C) increased with tissue nitrogen content, however, the Q(10) did not. Improved nutrition more than doubled stem basal respiration rate and increased branch basal respiration by 38%. Exponential equations were developed to model stem and branch respiration as a function of cambium temperature and tissue nitrogen content. We conclude that failure to account for tissue nitrogen effects on respiration rates will result in serious errors when estimating annual maintenance costs. PMID- 12651295 TI - Histological response of resistant and susceptible white spruce to simulated white pine weevil damage. AB - The traumatic wound response of families of white spruce, Picea glauca (Moench) Voss, resistant or susceptible to the white pine weevil, Pissodes strobi (Peck), were compared after simulated weevil damage. Leaders from 331 trees were wounded just below the apical bud in the spring, coinciding with the natural time of weevil oviposition. A portable 1-mm diameter drill was used to drill 24 holes per leader. Leaders were removed in the fall and examined for evidence of traumatic resin canal formation. Drilled trees had a traumatic wound response 8 times greater than that of undrilled trees; however, undrilled trees also formed some resin canals in response to unknown causes. In the drilled trees, the traumatic wound response extended into the lower part of the leader, where it could possibly affect older larvae. Trees from resistant families responded with greater intensity than trees from susceptible families, by producing multiple rings of traumatic resin canals. Trees from resistant families also responded more rapidly than trees from susceptible families based on number of cells to the first ring of traumatic resin canals. Trees from some resistant families exhibited no traumatic resin canal formation, showing considerable within-family variation and suggesting that other resistance mechanisms might be important. In the year after drilling, there was a reduction in tree diameter growth and trees suffered a reduction in constitutive resin canals in the bark, which suggests some energetic cost of traumatic resin production. There was no indication that the extent of constitutive defenses, as measured by density of cortical resin canals before wounding, was related to the ability to produce traumatic resin canals. Screening trees based on their capacity to produce traumatic resin canals may be useful in selecting genotypes resistant to white pine weevil. PMID- 12651296 TI - Developmental factors responsible for heterosis in aspen hybrids (Populus tremuloides x P. tremula. AB - Juvenile growth and bud set phenology were analyzed to study the biological basis of heterosis (hybrid vigor) in interspecific hybrids of Populus tremuloides Michx. (T) and P. tremula L. (Ta). Growth, measured as seedling volume index, was significantly higher for each of the two reciprocal interspecific crosses, T x Ta and Ta x T, than for the T x T intraspecific cross. Broad-sense heritabilities were 2-6 times larger than narrow-sense heritabilities for growth and shoot components in the T x T intraspecific cross, suggesting an important role for dominance or overdominance in aspen growth. Previous genetic analyses have indicated that hybrid vigor may be the result of overdominance at several key loci each with an allele inherited from each of parental species. Internode length and leaf number contributed substantially to the heterosis of stem volume, but their effects on heterosis differed between the T x Ta and Ta x T hybrids. In T x Ta seedlings, heterosis of stem volume was attributed to a high diameter growth rate, whereas in Ta x T seedlings heterosis of stem volume was probably the result of delayed bud set resulting in a longer duration of height growth. In addition to internode number and length and leaf number, other morphological or physiological components might affect heterosis, for example, extended leaf retention. PMID- 12651297 TI - Changes in the relationship between tree size and aboveground respiration in field-grown hinoki cypress (Chamaecyparis obtusa) trees over three years. AB - Respiration measurements of aerial parts of 18-year-old hinoki cypress (Chamaecyparis obtusa (Sieb. et Zucc.) Endl.) trees were made under field conditions over three years to study changing relationships with tree age between respiration and phytomass, phytomass increment, and leaf mass. The relationship between annual respiration (r(a)) and phytomass (w(T)) was approximated by a proportional function (r(a) = aw(T)), where the proportional constant (a) decreased year by year. The effect of time on the relationship between annual respiration and phytomass of each sample tree was fitted by a power function. Respiration of the tree suppressed by the canopy decreased year by year, but respiration of the other trees increased slightly with age. The relationship between annual respiration and leaf mass was also approximated by a generalized power function. Excluding the suppressed tree, the relationship between annual respiration (r(a)) and the annual increment of aboveground phytomass (Deltaw(T)) was described by a proportional function (r(a) = 2.27Deltaw(T)), where the proportional constant, 2.27, was independent of sample tree and year, indicating that about 2.3 times of the annual aboveground phytomass increment equivalent was respired annually. For any tree, the time constant relationships between annual respiration and leaf mass and phytomass increment for different-sized trees were similar to the corresponding time continuum relationships. In contrast, the time continuum relationship between annual respiration and phytomass differed from the time constant relationship, indicating that respiration of less active woody tissue contributed significantly to aboveground respiration. Based on the relationship between tree size and annual respiration, annual aboveground stand respiration was estimated to be 25.0, 26.9, and 25.8 Mg(dm) ha(-1) year(-1) for the three consecutive years, respectively, and the corresponding aboveground stand biomass was 60.0, 69.0, and 76.8 Mg(dm) ha(-1). PMID- 12651298 TI - Seasonal changes in leaf nitrogen pools in two Salix species. AB - Leaf nitrogen distribution pattern was studied four times during the growing season in a 2-year-old Salix viminalis L. and Salix dasyclados Wimm. plantation in Estonia. We measured the vertical distributions of leaf nitrogen concentration, dry mass, leaf area and light environment (as fractional transmission of diffuse irradiance, a(d)) in the canopy. The light-independent nitrogen pool was evaluated as the intercept of the leaf nitrogen concentration versus a(d) relationship, and the nondegradable nitrogen pool was evaluated as the nitrogen remaining in abscised leaves. A strong vertical gradient of mass based leaf nitrogen concentration was detected at the beginning of the growing season, and decreased steadily during canopy development. This decline had at least three causes: (1) the amount of nitrogen in the foliage was larger at the beginning of the growing season than at the end of the growing season, probably because of pre-existing root systems; (2) with increasing leaf area index (LAI) during the growing season, the proportion of leaf nitrogen in total canopy nitrogen that could be redistributed (light-dependent nitrogen pool) decreased; and (3) the photosynthetic photon flux density gradient inside the canopy changed during the season, most probably because of changes in leaf area and leaf angle distributions. Total canopy nitrogen increased almost proportionally to LAI, whereas the light-dependent nitrogen pool had a maximum in August. Also, the proportion of the light-dependent nitrogen pool in the total canopy nitrogen decreased steadily from 65.2% in June to 17.2% in September in S. dasyclados and from 63.3 to 15.1% in S. viminalis. The degradable nitrogen pool was always bigger than the light-dependent nitrogen pool. PMID- 12651299 TI - Accumulation of flavonoids and related compounds in birch induced by UV-B irradiance. AB - A growth chamber experiment was conducted to examine the effects of UV-B exposure (4.9 kJ m(-2) day(-1) of biologically effective UV-B, 280-320 nm) on shoot growth and secondary metabolite production in Betula pendula (Roth) and B. resinifera (Britt.) seedlings originating from environments in Finland, Germany and Alaska differing in solar UV-B radiation and climate. Neither shoot growth nor the composition of secondary metabolites was affected by UV-B irradiance, but the treatment induced significant changes in the amounts of individual secondary metabolites in leaves. Leaves of seedlings exposed to UV-B radiation contained higher concentrations of several flavonoids, condensed tannins and some hydroxycinnamic acids than leaves of control seedlings that received no UV-B radiation. At the population level, there was considerable variation in secondary metabolite responses to UV-B radiation: among populations, the induced response was most prominent in Alaskan populations, which were adapted to the lowest ambient UV-B radiation environment. I conclude that solar UV-B radiation plays an important role in the formation of secondary chemical characteristics in birch trees. PMID- 12651300 TI - Growth and development of the arborescent cactus Stenocereus queretaroensis in a subtropical semiarid environment, including effects of gibberellic acid. AB - In Stenocereus queretaroensis (Weber) Buxbaum, an arborescent cactus cultivated in Jalisco, Mexico, for its fruits but studied here in wild populations, stem extension occurred in the autumn at the beginning of the dry season, flowering and fruiting occurred in the spring at the end of the dry season, and new roots grew in the summer during the wet season. The asynchrony of vegetative and reproductive growth reduces competitive sink effects, which may be advantageous for wild populations growing in infertile rocky soils. Seasonal patterns of sugars in the roots and especially the stems of S. queretaroensis were closely related to the main phenological stages, becoming lower in concentration during periods of major stem extension. Cessation of stem extension occurred in 100-year old plants for which injection of GA(3) reinitiated such growth. Isolated chlorenchyma cylinders had maximum extension in a bathing solution containing 0.1 &mgr;M gibberellic acid. PMID- 12651301 TI - Thigmomorphogenesis: a dose response to flexing in Ulmus americana seedlings. AB - Six-week-old half-sib seedlings of Ulmus americana L. were subjected to different amounts of flexure daily for 3 weeks under controlled greenhouse conditions. The daily flexure treatments were: no flexing in a staked stem, minimal flexing in a non-staked stem, and five, 10, 20, 40, and 80 flexures. Seedling height and diameter growth and average leaf area were determined before and after the treatments. The ratio of the change in height growth (DeltaH) to the change in diameter growth (DeltaD; (DeltaH:DeltaD)) before and after the 3-week treatments were calculated. At the end of the 3-week experiment, staked seedlings were significantly taller and had smaller stem diameters than all of the flexed seedlings. Height growth tended to decrease exponentially with increased flexure, with significant differences between the extremes of treatment. All of the flexure treatments significantly increased stem diameter compared to staked seedlings. The DeltaH:DeltaD ratio exhibited an exponential function in response to increased flexure. Average leaf area decreased with increased flexure, and seedlings in the 40x and 80x flexure treatments had significantly less leaf area than seedlings in all of the other treatments. These data are similar to the dose responses previously observed in herbaceous species. The finding that trees exhibit greater sensitivity to low doses of flexure than to high doses of flexure indicates that slight exposure to wind may result in a large initial alteration in stem morphology, producing a thigmomorphogenetic effect. Trees will continue to respond to increasing amounts of mechanical stress, but at an exponentially declining rate. Declining leaf areas in response to increasing amounts of mechanical stress may result in a decrease in available photosynthate, resulting in a tree of smaller stature compared to trees exposed to lower amounts of mechanical loading. PMID- 12651302 TI - Detection and quantification of changes in membrane-associated calcium in red spruce saplings exposed to acid fog. AB - Five-year-old red spruce saplings (Picea rubens Sarg.) were exposed to either (1) acid fog consisting of a mixture of H(2)SO(4) and HNO(3) adjusted to pH 2.5, (2) distilled-water fog at pH 5.6, or (3) no fog (dry control) for 3.5 hours per day, five times a week during the 1996 and 1997 growing seasons. The effect of fog on cell membrane-associated calcium (mCa) of leaf mesophyll cells was investigated with the fluorescence probe chlortetracycline (CTC). In both years, mean mCa concentrations were significantly less in needles exposed to acid fog than in needles exposed to distilled-water fog or in untreated needles. In 1997, acid-fog treatment resulted in 25 and 12% reductions in mCa in current-year needles, and 18 and 15% reductions in 1-year-old needles, compared with untreated needles and needles exposed to distilled-water fog, respectively, indicating that acid deposition induced calcium leaching from the membranes of photosynthetic mesophyll cells. Exposure to distilled-water fog also led to reductions in mCa in young needles, suggesting that water films on needle surfaces can induce losses by diffusion between the needle interior and surface. Consistent with the chamber studies, field data obtained from red spruce trees at two sites in Maine showed that low mCa concentrations in needles were associated with exposure to acid fog. PMID- 12651303 TI - Kinetics of leaf temperature fluctuation affect isoprene emission from red oak (Quercus rubra) leaves. AB - Because the rate of isoprene (2-methyl-1,3-butadiene) emission from plants is highly temperature-dependent, we investigated natural fluctuations in leaf temperature and effects of rapid temperature change on isoprene emission of red oak (Quercus rubra L.) leaves at the top of the canopy at Harvard Forest. Throughout the day, leaves often reached temperatures as much as 15 degrees C above air temperature. The highest temperatures were reached for only a few seconds at a time. We compared isoprene emission rates measured when leaf temperature was changed rapidly with those measured when temperature was changed slowly. In all cases, isoprene emission rate increased with increasing leaf temperature up to about 32 degrees C and then decreased with higher temperatures. The temperature at which isoprene emission rates began to decrease depended on how quickly measurements were made. Isoprene emission rates peaked at 32.5 degrees C when measured hourly, whereas rates peaked at 39 degrees C when measurements were made every four minutes. This behavior reflected the rapid increase in isoprene emission rate that occurred immediately after an increase in leaf temperature, and the subsequent decrease in isoprene emission rate when leaf temperature was held steady for longer than 20 minutes. We concluded that the observed temperature response of isoprene emission rate is a function of measurement protocol. Omitting this parameter from isoprene emission models will not affect simulated isoprene emission rates at mild temperatures, but can increase isoprene emission rates at high temperatures. PMID- 12651304 TI - Flood tolerance of four tropical tree species. AB - Many seasonally flooded habitats in the tropics are dominated by one or a few tree species. We tested the hypothesis that the inability to tolerate flooding restricts most species from becoming established in flood-prone habitats. We compared morphological and physiological responses to flooding in seedlings of Prioria copaifera Griseb., a species that forms monodominant stands in seasonally flooded habitats, and in three species confined to flood-free sites; namely, Calophyllum longifolium Willd., Virola surinamensis Aubl. and Gustavia superba (H.B.K.) Berg. Flooding reduced photosynthesis at Day 45 in all species by 10 30%. By Day 90, photosynthesis returned to the control rate in Prioria, but not in the other species. Flooding reduced stomatal conductance by 25-35% in all species except Calophyllum, and it reduced leaf area growth by 44% in Virola, but not in the other species. All species survived 90 days of flooding without mortality, leaf chlorosis, leaf necrosis, or leaf abscission. Flooding reduced root:shoot ratio significantly in Gustavia and Calophyllum, but not in the other species, and it reduced maximum root depth by 29% in Prioria, but by 61% or more in the species from flood-free habitats. PMID- 12651305 TI - Growth adaptation of leaves and internodes of poplar to irradiance, day length and temperature. AB - The adaptation of absolute growth rate of leaves and internodes of Populus euramericana (Dode) Guinier cv. 'Robusta' to irradiance and day length proceeds by increases in the volume of the apex. Diameter and height of the apex increase linearly with time, resulting in linear increases in rates of leaf initiation and stem height growth that can be described by an acceleration factor. The acceleration factor is proportional to day length. The relationship between the acceleration factor and irradiance is curvilinear and saturates at irradiances above about 300 W m(-2). Absolute stem height growth rate is the product of mean relative growth rate in stem height and the length of the growing part of the stem. The temperature-dependent growth pattern of each individual leaf or internode reflects a specific relationship between its relative growth rate and organ age that is independent of irradiance and plant age; however, it is dependent on day length during the primordial phase. The constancy of the growth patterns and the correlation between leaf length and leaf initiation rate indicate that growth of primordia is predetermined in the apex, presumably by the precisely structured vascular system. PMID- 12651306 TI - Nitrogen and potassium effects on xylem cavitation and water-use efficiency in poplars. AB - Effects of N and K nutrition on drought and cavitation resistance were examined in six greenhouse-grown poplar clones: Populus trichocarpa (Torr. & Gray) and its hybrids with P. deltoides Bartr. and P. euramericana (Dole) Guinier, before and after preconditioning to water stress. Both tendency to cavitate and water-use efficiency (WUE) increased when N supply was increased, whereas K supply had little impact on cavitation. Mean xylem vessel diameters increased from 36.6 &mgr;m at low-N supply to 45.2 &mgr;m at high-N supply. Drought-hardy clones, which were relatively resistant to cavitation, had the smallest mean vessel diameters. Vulnerability to cavitation had a weakly positive relationship with vessel diameter, and a negative correlation with transpiration. Drought hardening offered no protection against cavitation in a subsequent drought. Under drought conditions, increasing N supply increased leaf loss and decreased water potentials, whereas increasing K supply decreased leaf loss. Drought-resistant clones exhibited similar WUE to drought-susceptible clones, but had smaller, more numerous stomata and greater leaf retention under drought conditions. PMID- 12651307 TI - Responses of Scots pine (Pinus sylvestris) seedlings grown in different nutrient regimes to changing root zone temperature in spring. AB - We examined effects of nutrient availability and changing root zone temperature (RZT) on growth, gas exchange and plasma membrane H(+)-ATPase (PM-ATPase) activity of roots of 1-year-old Scots pine (Pinus sylvestris L.) seedlings during spring flushing. The 6-week growth-chamber experiment was carried out in hydroponic cultures that supplied the seedlings with low (0.5 mM N) or high (3 mM N) nutrient concentration and two rates of increase in RZT were simulated: slow warming (SW-treatment) and fast warming (FW-treatment). Air temperature, humidity, and light conditions were similar in all treatments. Growth of roots and shoots was retarded at low RZT, and fresh mass increment of roots was closely correlated with RZT sum. High nutrient availability increased nitrogen concentrations of needles and stems, but only at RZTs >13 degrees C. Low RZT and low availability of nutrients suppressed gas exchange of the seedlings. Real PM ATPase activity was highly dependent on RZT. At high RZTs, real PM-ATPase activity was affected by nutrient availability but this effect was related to root growth. We conclude that, under conditions of high nutrient availability, Scots pine seedlings can compensate for the suppressive effects of long-term exposure to low RZT by rapidly accelerating growth, gas exchange and root metabolism, but only when RZT has increased above a threshold value, which was 13 degrees C in this study. PMID- 12651308 TI - Characterization of radiation regimes in nonrandom forest canopies: theory, measurements, and a simplified modeling approach. AB - We used field measurements and Monte Carlo simulations of canopy gap-size distribution and gap fraction to examine how beam radiation interacts with clumped boreal forest canopies of aspen (Populus tremuloides Michx.), black spruce (Picea mariana (Mill.) B.S.P.) and jack pine (Pinus banksiana Lamb.). We demonstrate that the Beer-Lambert law can be modified to accommodate transmission of radiation through a clumped forest canopy as a function of path length or sun zenith angle. Multiband Vegetation Imager (MVI) measurements and Monte Carlo simulations showed that values of the zenith element clumping index (Omega(e)(0)) are typically between 0.4 and 0.5 in jack pine and black spruce and 0.65 in aspen. Estimates of LAI obtained from MVI measurements of the canopy gap fraction and adjusted for canopy clumping and branch architecture yielded LAI values of 3.0 in jack pine, 3.3 in aspen, and about 6.0 in black spruce. These LAI estimates were within 10-25% of direct measurements made at the same sites. Data obtained with the MVI, along with numerical simulations, demonstrated that assumptions of random foliage distributions in boreal forests are invalid and could yield erroneous values of LAI measured by indirect techniques and false characterizations of atmosphere-biosphere interactions. Monte Carlo simulations were used to develop a general equation for beam radiation penetration as a function of zenith angle in clumped canopies. The essential measurements included stem spacing, crown diameter, crown depth, and within-crown gap fraction. PMID- 12651309 TI - Photosynthetic responses to phosphorus nutrition in two-year-old maritime pine seedlings. AB - We analyzed processes limiting photosynthesis in two-year-old, container-grown Pinus pinaster Ait. seedlings subjected to phosphorus (P) deficiency. After withholding P for 3 months, seedlings were supplied P at four relative addition rates (0, 0.005, 0.01 and 0.02 day(-1)) in a nutrient recycling system. At Weeks 12 and 22, responses of photosynthesis to CO(2) and irradiance were measured and the following parameters derived: maximal velocity of carboxylation by Rubisco, V(m); apparent quantum efficiency of electron transport, alpha maximal electron transport rate, J(m); stomatal conductance and relative stomatal limitation of photosynthesis. At Week 22, these measurements were combined with concurrent measurements of chlorophyll fluorescence to determine the quantum yield of PSII, and a theoretical partitioning of total light-driven linear electron flow between fractions used to regenerate carboxylated and oxygenated RuBP. After 12 weeks of treatment, needle P concentrations ranged from 0.04 to 0.15 x 10(-2) g g(DW) ( 1), and then remained constant until Week 22. Values of J(m), alpha and V(m) increased with increasing needle P concentration (from 30 to 133 &mgr;mol m(-2) s(-1), 0.02 to 0.25 mol mol(-1) and 13 to 78 &mgr;mol CO(2) m(-2) s(-1) at the lowest and highest needle P concentrations, respectively). Under ambient conditions, net assimilation rates in P-deficient seedlings were limited by V(m) under saturating irradiance, and by J(m) under limiting irradiance, but not by triose-P regeneration. There was no detectable change in the partitioning of total light-driven linear electron flow between the fractions used for carboxylation and oxygenation. Predawn photochemical efficiency of PSII was significantly reduced in seedlings with low P concentrations. Although stomatal conductance tended to decrease with decreasing needle P concentration, relative stomatal limitation was not significantly affected. At Week 22, there was an attenuation of the effects of P nutrition on V(m) and an increase in alpha and J(m) that was probably related to cessation of growth and the seasonal decline in natural irradiance. PMID- 12651310 TI - Partitioning of soil water among tree species in a Brazilian Cerrado ecosystem. AB - Source water used by woody perennials in a Brazilian savanna (Cerrado) was determined by comparing the stable hydrogen isotope composition (deltaD) of xylem sap and soil water at different depths during two consecutive dry seasons (1995 and 1996). Plant water status and rates of water use were also determined and compared with xylem water deltaD values. Overall, soil water deltaD decreased with increasing depth in the soil profile. Mean deltaD values were -35 per thousand for the upper 170 cm of soil and -55 per thousand between 230 and 400 cm depth at the end of the 1995 dry season. Soil water content increased with depth, from 18% near the surface to about 28% at 400 cm. A similar pattern of decreasing soil water deltaD with increasing depth was observed at the end of the 1996 dry season. Patterns consistent with hydraulic lift were observed in soil profiles sampled in 1995 and 1997. Concurrent analyses of xylem and soil water deltaD values indicated a distinct partitioning of water resources among 10 representative woody species (five deciduous and five evergreen). Among these species, four evergreen and one deciduous species acquired water primarily in the upper soil layers (above 200 cm), whereas three deciduous and one evergreen species tapped deep sources of soil water (below 200 cm). One deciduous species exhibited intermediate behavior. Total daily sap flow was negatively correlated with xylem sap deltaD values indicating that species with higher rates of water use during the dry season tended to rely on deeper soil water sources. Among evergreen species, minimum leaf water potentials were also negatively correlated with xylem water deltaD values, suggesting that access to more readily available water at greater depth permitted maintenance of a more favorable plant water status. No significant relationship between xylem water deltaD and plant size was observed in two evergreen species, suggesting a strong selective pressure for small plants to rapidly develop a deep root system. The degree of variation in soil water partitioning, leaf phenology and leaf longevity was consistent with the high diversity of woody species in the Cerrado. PMID- 12651311 TI - Impact of snow cover on photoinhibition and winter dessication in evergreen Rhododendron ferrugineum leaves during subalpine winter. AB - Effects of winter snow cover on photoinhibition and possible interactions with winter desiccation were investigated in situ in an evergreen subalpine woody species, Rhododendron ferrugineum L., at the alpine timberline (1950 m a.s.l.). Timing and duration of complete snow cover markedly influenced potential efficiency of photosystem II (PSII; F(v) /F(m)). Lack of snow cover led to severe but mostly reversible photoinhibition with F(v)/F(m) values as low as 0.05. Complete snow cover immediately stopped further reductions in PSII efficiency. Snow cover promoted recovery from photoinhibition, but only if, in addition to shading by snow, plants were exposed to nonfreezing temperatures close to 0 degrees C. The F(v)/F(m) ratio was closely related to minimum leaf temperatures because both photoinhibition and recovery from photoinhibition were strongly influenced by temperature. The period without major reductions in PSII efficiency lasted for only two months, reflecting the extremely short growing period in the subalpine environment. Compared with complete snow cover, incomplete snow cover led to significantly higher water losses as well as lower dehydration tolerance, because both osmotic adjustment and changes in turgor maintenance capacity were significantly reduced. Interactions between photoinhibition and winter desiccation were masked by the direct effects of freezing temperatures. However, both photoinhibition and winter desiccation are closely linked and occur together under field conditions in this evergreen subalpine woody species. PMID- 12651312 TI - Thermally induced changes in shoot morphology of Hancornia speciosa microcultures: evidence of mediation by ethylene. AB - A shoot microculture protocol for the tropical fruit tree Hancornia speciosa Gom. was developed based on high temperature stimulation of axillary branching. An increase in the culture temperature from 25 to 31 degrees C (continuous over a 4 week period) stimulated the elongation of newly subcultured micro-shoots. Increases in culture temperature from 31 to 35 degrees C suppressed elongation but induced branching of all shoots. Increasing temperature to 37 degrees C reduced both shoot elongation and branching severely. Thermoperiodic regimes involving daily exposure for 8 or 16 h at 35 degrees C followed by 31 degrees C for the remainder of the day were only moderately effective in stimulating branching. However, maintaining cultures at 35 degrees C for 2 weeks followed by growth at a constant temperature of 31 degrees C led to vigorous branching. We hypothesize that thermally induced branching is mediated by a reduction in ethylene biosynthesis. In cultures of nodal segments grown at 31 degrees C, ethylene evolution peaked 12 to 16 days after subculture, and then gradually decreased until the end of the culture cycle. Compared with cultures held at 31 degrees C, those grown at 35 degrees C showed an earlier peak and a reduced rate of ethylene evolution throughout most of the culture cycle, and less 1 aminocyclopropane-1-carboxylate (ACC) synthase mRNA transcript and ACC oxidase activity. An inhibitor of ethylene biosynthesis (L-(2-aminoethoxyvinyl)-glycine) at 4.5 &mgr;M mimicked the effect of elevated temperature (35 degrees C) in stimulating lateral branching. The branching pattern of shoots grown at 35 degrees C in the presence of 1 &mgr;l l(-1) ethylene gas resembled that of shoots grown at 31 degrees C. We conclude that a reduction in ethylene evolution has a role in thermal induction of branching in H. speciosa. Heat induced release of axillary buds may be useful in the microculture of trees with strong apical dominance that cannot be overcome by cytokinin treatment. PMID- 12651313 TI - Interactive effects of leaf age and self-shading on leaf structure, photosynthetic capacity and chlorophyll fluorescence in the rain forest tree, Dryobalanops aromatica. AB - In the tropical canopy tree, Dryobalanops aromatica Gaertn. f., upper-canopy leaves (UL) develop under sunlit conditions but are subjected to self-shading within the crown as they age. In contrast, lower-canopy leaves (LL) are exposed to uniform dim light conditions throughout their life span. By comparing leaf morphology and physiology of UL and LL, variations in leaf characteristics were related to leaf age and self-shading. Mass-based chlorophyll (chl) concentration and the chlorophyll/nitrogen (chl/N) ratio were lower and the chl a/b ratio was higher in UL than in LL. In UL, the chl/N ratio gradually increased and the chl a/b ratio gradually decreased with leaf aging, whereas these ratios remained unchanged with leaf age in LL. The effective quantum yield of photosystem II (PSII) (DeltaF/F(m)') at a given irradiance remained unchanged with leaf age in LL, whereas DeltaF/F(m)' changed with leaf age in UL. These data indicate N reallocation within the leaves from carbon fixation components to light harvesting components and a dynamic regulation of photochemical processes of PSII in response to increased self-shading of UL. Despite the difference in light environment with leaf age between UL and LL, maximum photosynthetic rates and nitrogen-use efficiency decreased with leaf aging in both UL and LL. Constancy in the chl/N ratio with leaf age in LL indicated that the decrease in photosynthetic capacity was caused by effects other than shading, such as leaf aging. We conclude that N reallocation and acclimation of PSII to self-shading occurred even in mature leaves, whereas the change in photosynthetic capacity with leaf age was more conservative. PMID- 12651314 TI - Fall frost resistance in willows used for biomass production. I. Characterization of seasonal and genetic variation. AB - Fast-growing willow clones (six clones of Salix viminalis L. and one clone each of S. viminalis x S. schwerenii E. Wolf and S. dasyclados Wimm.) were compared with respect to growth rhythm and frost hardening in the fall. Frost resistance of stem tissues was assessed by controlled freezing followed by analysis of chlorophyll fluorescence and scoring of visible cambial discoloration. The fluorescence method proved superior to scoring based on visible cambial discoloration because it was more rapid and less subjective, but needed calibration against cambial damage. Frost hardening in mature parts of stems did not start until growth cessation was initiated in the shoot apices, irrespective of whether growth cessation occurred early or late in the fall. Frost resistance varied because of clonal variations in: (1) pre-hardening frost resistance; (2) timing of growth cessation and hence start of frost hardening; and (3) rate of frost hardening. Compared with coastal and southern clones, continental and northern clones started hardening earlier, and a continental clone proceeded through hardening more rapidly at a given temperature. A cross between a continental and coastal clone was intermediate in timing. The pre-hardening frost resistance, however, was unrelated to both growth and frost hardening characteristics. PMID- 12651315 TI - Fall frost resistance in willows used for biomass production. II. Predictive relationships with sugar concentration and dry matter content. AB - The accumulation of sugars and dry matter in stems in fall was examined in relation to frost hardening in eight willow clones (six clones of Salix viminalis L. and one clone each of S. viminalis x S. schwerenii E. Wolf and S. dasyclados Wimm.). Evidence is presented that three sources of variation in fall frost resistance among the eight clones could be assessed from an analysis of stem composition. First, the pre-hardening value of frost resistance could be assessed from the total sugar concentration. Second, the start of induction of apical growth cessation and hence frost hardening could be distinguished by a stepwise increase in sucrose-to-glucose ratio. Third, the progress of frost hardening during its first phase could be followed from a proportional rise in total sugar concentration and, even more accurately, from a proportional rise in dry-to-fresh weight ratio. In contrast, the second phase of frost hardening was largely uncoupled from sugar and dry matter accumulation. Raffinose and sucrose accumulation seemed to be under differential environmental controls. Sucrose accumulation started with the initiation of growth cessation controlled by photoperiod, whereas raffinose accumulation started with falling temperatures later on. Starch reserves that built up in stems in early fall were partially mobilized later on to support sugar accumulation. In contrast to stems, leaves did not exhibit a preferential accumulation of sucrose in fall. PMID- 12651316 TI - Influence of rate of change in stomatal conductance to fluctuating irradiance on estimates of daily water use by Pinus taeda leaves. AB - Pinus taeda L. stomata respond slowly to changes in irradiance. Because incident irradiance on a leaf varies constantly, the rate of change in stomatal conductance to fluctuating irradiance may have a large effect on plant water use. We estimated total daily water use of Pinus taeda foliage for 10 days with very different irradiance patterns, assuming that rates of stomatal opening and closing were similar. To determine how the most extreme imbalance in rates of stomatal opening and closing affects estimates of water use, we also estimated total daily water use assuming instantaneous stomatal opening and a realistic rate of stomatal closing. Total daily water use was calculated by summing estimates of transpiration based on irradiance and vapor pressure deficit measured every minute at locations atop and within a canopy. Estimates of total daily water use calculated on the basis of realistic rates of stomatal opening and closing were similar to estimates calculated assuming instantaneous stomatal change (mean difference between methods of calculation was less than 0.2%). Estimates of total daily water use assuming instantaneous stomatal opening and a realistic rate of closing differed from estimates of total daily water use based on similar rates of stomatal opening and closing. The discrepancy was greater within the canopy (mean difference 6%) than at the top of the canopy (mean difference 1%). Calculation of mean daily conductance from mean daily irradiance, without accounting for minute-by-minute variations in irradiance, resulted in overestimations of daily stomatal conductance (13% mean error) and the magnitude of the error was directly related to the variation in irradiance for that day. We conclude that, provided variation in irradiance is accounted for, rates of stomatal opening and closing have little effect on estimates of daily water use. PMID- 12651317 TI - Corrected heat capacity of wood for sap flow calculations. AB - Values for the temperature-dependent heat capacity of wood were applied to correct the coefficient used to calculate sap velocity from heat pulse velocity, leading to values at 20 degrees C about 5% smaller than values calculated with the widely employed algorithm of Edwards and Warwick. Errors are larger at lower temperatures (7% at 10 degrees C) and so are more problematic at night and during cold periods. PMID- 12651318 TI - Photosynthetic induction responses to variable light under field conditions in three species grown in the gap and understory of a Fagus crenata forest. AB - Photosynthetic induction responses to abrupt increases in photon flux density (PFD) to 800 and 1500 &mgr;mol m(-2) s(-1) from either darkness or 100 &mgr;mol m(-2) s(-1) were examined in situ in leaves of Fagus crenata Blume, Daphniphyllum humile Maxim., and Acer rufinerve Siebold & Zucc. growing in a gap and the understory of an F. crenata forest. Among the species studied, F. crenata exhibited the highest assimilation rate (A(100)), stomatal conductance (g(s100)) at the background PFD of 100 &mgr;mol m(-2) s(-1), and A(100)/A(max) (A(max) = maximum assimilation rate), in both the gap and the understory. Time required for full induction depended on both background PFD and maximum PFD. The induction period was 2-4-fold shorter at a background PFD of 100 &mgr;mol m(-2) s(-1) than in darkness. For the three understory species, time required to full induction was 2-3-fold longer when irradiance was increased from darkness to 800 &mgr;mol m(-2) s(-1) than when irradiance was increased from darkness to 1500 &mgr;mol m( 2) s(-1). Acer rufinerve showed higher initial stomatal conductance (g(s0)) and a shorter induction period in the understory than in the gap. Fagus crenata exhibited a similar g(s0) and induction period in both habitats. Daphniphyllum humile demonstrated lower g(s0) and a longer induction period in the understory than in the gap. These findings indicate that initial stomatal conductance is closely correlated with the photosynthetic induction response. We conclude that the photosynthetic induction response is affected by the light conditions experienced by plants before the sudden increase in irradiance and by the extent of the increase in irradiance. PMID- 12651319 TI - Diurnal changes in chlorophyll a fluorescence, CO(2)-exchange and organic acid decarboxylation in the tropical CAM tree Clusia hilariana. AB - Crassulacean acid metabolism (CAM) plants are dependent on the organic acids that accumulate overnight in the vacuoles as a source of CO(2) during the daylight deacidification period, when stomata are closed and high irradiances generally prevail. We performed an integrative analysis of diurnal changes in gas exchange, chlorophyll fluorescence parameters and organic acid decarboxylation to understand the adjustments in photochemical and non-photochemical processes during the different CAM phases in Clusia hilariana Schlecht., a dominant tree species in the sandy coastal plains of southeastern Brazil. A linear relationship was obtained between the quantum yields of photochemical and non-photochemical quenching, irrespective of the CAM phase and prevailing irradiance. Degradation of malic and citric acids during the midday stomatal closure period could lead to potential CO(2) fixation rates of 23 &mgr;mol m(-2) s(-1), whereas CO(2) losses, measured as CO(2) evolution, corresponded to about 3% of this value. Thus, decarboxylation of malate and citrate provided high internal CO(2) concentrations during phase III of CAM, even though the stomata were closed, allowing optimal utilization of light energy, as indicated by the non-saturating electron transport rates (ETR) in the light response curves, with highest rates of ETR occurring at midday in the diurnal curves. At the transition from phase III to IV of CAM, depletion of internal CO(2) sources and low stomatal conductances, which restricted the supply of exogenous CO(2), reduced the demand for photochemical energy to drive carbon assimilation. This was compensated by increases in thermal energy dissipation as indicated by higher rates of non-photochemical quenching, while high irradiances still prevailed. Shifts in the CAM phases and changes in protective thermal dissipation potential allowed C. hilariana to match changes in PPFD patterns for leaves of different orientations. Evidence that most of the decline in photochemical efficiency was probably related to the fast-relaxing component of non-photochemical quenching is provided by the high values of the quantum yield of photosystem II after 20 min of relaxation in darkness, and an almost complete recovery after sunset. These adjustments in photosynthetic machinery minimized the danger of photo-inhibition in C. hilariana, which is commonly found in fully exposed habitats. PMID- 12651320 TI - Localized graft incompatibility in pear/quince (Pyrus communis/Cydonia oblonga) combinations: multivariate analysis of histological data from 5-month-old grafts. AB - To characterize the structural events associated with incompatibility of graft development, we conducted a histological study of compatible and incompatible pear/pear and pear/quince grafts that had been grown for five months in a greenhouse. Multivariate analysis of histological data describing the structure of the graft union allowed us to discriminate between compatible and incompatible combinations before either macroscopic examination or qualitative microscopic examination differences between graft combinations became evident. The histological variables responsible for the discrimination between incompatible and compatible unions were related to three typical symptoms of graft incompatibility: bark discontinuity, which was the main feature; cambial dysfunction; and accumulation of starch in the scion. Little cell necrosis was observed at the interface of incompatible grafts at the 5-month stage of graft development. Multivariate analysis of histological data provides a new tool for studying early structural events resulting from the graft incompatibility response and for diagnosing early graft incompatibility. PMID- 12651321 TI - Mineral nutrition, resin flow and phloem phytochemistry in loblolly pine. AB - Southern pine beetles and associated pathogenic fungi represent the largest biotic threat to pine forests in the southeastern USA. The two primary defensive mechanisms of the tree to the beetle-fungal complex are the primary oleoresin flow and the concentrations of preformed and induced secondary compounds. We compared oleoresin flow and concentrations of phloem nutrients, soluble sugars, starch, total phenolics and proanthocyanidins in Pinus taeda L. trees in fertilized and control plots in the Sandhills region of North Carolina. Four blocks of 10 trees per treatment were sampled on five dates from May to November 1995. Phloem nitrogen and potassium concentrations were elevated in trees on fertilized plots, whereas phloem calcium concentrations were decreased. Fertilization significantly enhanced (10-20%) concentrations of phloem phenolics and proanthocyanidins. In contrast, phloem soluble sugars and starch concentrations were up to 30% lower in fertilized trees than in control trees. Increased phenolic concentrations and lower nonstructural carbohydrates should correlate with reduced tissue palatability and decreased pathogen susceptibility in fertilized trees; however, resin flows were significantly lower (30-100%) in fertilized trees compared with control trees, which may facilitate pine bark beetle establishment. Furthermore, fertilization-induced increases in phloem nitrogen concentration may be more important than tissue carbohydrate or phenolic content in determining tissue palatability. PMID- 12651322 TI - Seasonal changes in photosynthesis of eight savanna tree species. AB - Seasonal variations in carbon assimilation of eight tree species of a north Australian tropical savanna were examined over two wet seasons and one dry season (18 months). Assimilation rates (A) in the two evergreen species, Eucalyptus tetrodonta F. Muell. and E. miniata A. Cunn. ex Schauer, were high throughout the study although there was a 10-20% decline in the dry season compared with the wet season. The three semi-deciduous species (Erythrophleum chlorostachys (F. Muell.) Baillon, Eucalyptus clavigera A. Cunn. ex Schauer, and Xanthostemon paradoxus F. Muell.) showed a 25-75% decline in A in the dry season compared with the wet season, and the deciduous species (Terminalia ferdinandiana Excell, Planchonia careya (F. Muell.) Kunth, and Cochlospermum fraseri Planchon) were leafless for several months in the dry season. Generally, the ratio of intercellular CO(2) concentration to ambient CO(2) concentration (C(i):C(a)) was larger in the wet season than in the dry season, indicating a smaller stomatal limitation of photosynthesis in the wet season compared with the dry season. In all species, the C(i):C(a) ratio and A were essentially independent of leaf-to-air vapor pressure difference (LAVPD) during the wet season, but both parameters generally declined with increasing LAVPD in the dry season. The slope of the positive correlation between A and transpiration rate (E) was less in the wet season than in the dry season. There was no evidence that high E inhibited A. Instantaneous transpiration efficiency was lowest in the wet season and highest during the dry season. Nitrogen-use efficiency (NUE) was higher in the wet season than in the dry season because the decline in A in the dry season was proportionally larger than the decline in foliar nitrogen content. In the wet season, evergreen species exhibited higher NUE than semi-deciduous and deciduous species. In all species, A was linearly correlated with specific leaf area (SLA) and foliar N content. Foliar N content increased with increasing SLA. All species showed a decline in midday leaf water potential as the dry season progressed. Dry season midday water potentials were lowest in semi-deciduous species and highest in the deciduous species, with evergreen species exhibiting intermediate values. PMID- 12651323 TI - Seasonal variations in leaf area index, leaf chlorophyll, and water content; scaling-up to estimate fAPAR and carbon balance in a multilayer, multispecies temperate forest. AB - Seasonal differences in phenology between coniferous and deciduous tree species need to be considered when developing models to estimate CO(2) exchange in temperate forest ecosystems. Because seasonal variations in CO(2) flux in temperate forests are closely correlated with plant phenology, we quantified the phenology of forest species in a multilayered forest with patches of Scots pine (Pinus sylvestris L.) and oak (Quercus robur L.) in Brasschaat, Belgium. A scaling-up modeling approach was developed to simulate reflectance at the leaf and canopy scales over a one-year cycle. Chlorophyll concentration, water content, specific leaf area and leaf area index of the forest species were measured throughout an entire year (1997). Scaling-up from the leaf to canopy was achieved by linking the PROSPECT and SAIL models. The result is the annual progression of the fraction of absorbed photosynthetically active radiation (fAPAR) in a 1 km(2) forest area, which can be directly related to high resolution, remotely sensed data. PMID- 12651324 TI - Potential errors in measurement of nonuniform sap flow using heat dissipation probes. AB - The empirical calibration of Granier-type heat dissipation sap flow probes that relate temperature difference (DeltaT) to sap velocity (v) was reevaluated in stems of three tropical tree species. The original calibration was confirmed when the entire heated probe was in contact with conducting xylem, but mean v was underestimated when part of the probe was in contact with nonconducting xylem or bark. Analysis of the effects of nonuniform sap velocity profiles on heat dissipation estimates showed that errors increased as v and the proportion of the probe in nonconducting wood increased. If half of a 20-mm probe is in sapwood with a v of 0.15 mm s(-1) and the other half is in nonconducting wood, then mean v for the whole probe can be underestimated by as much as 50%. A correction was developed that can be used if the proportion of the probe in nonconducting wood is known. Even with the entire heated probe in contact with conducting xylem, v would be underestimated when radial velocity gradients are present. In this case, the error would be smaller except when velocity gradients are very steep, as can occur in species with ring-porous wood anatomy. Errors occur because the relationship between DeltaT and v is nonlinear. Mean DeltaT along the probe is therefore not a measure of mean v, and users of heat dissipation probes should not assume that v is integrated along the length of the probe. The same type of error can occur when DeltaT is averaged through time while v is changing, but the error is small unless there are sudden, step changes between zero and high sap velocity. It is recommended that relatively short probes (20 mm or less) be used and that probes longer than the depth of conducting sapwood be avoided. Multiple probes inserted to a range of depths should be used in situations where steep gradients in v are expected. If these conditions are met, heat dissipation probes remain useful and widely applicable for measuring sap flow in woody stems. PMID- 12651325 TI - Relationships between gas exchange and carbon isotope discrimination of Sitka x interior spruce introgressive genotypes and ribosomal DNA markers. AB - Effects of hybridization on physiological performance were investigated in seven natural introgressive populations and one F(1) population of Sitka (Picea sitchensis (Bong.) Carr.) x interior spruce (P. glauca (Moench) Voss. x P. engelmannii Parry ex Engelm.). Each population was represented by a Sitka interior spruce ribosomal DNA (Si rDNA) index that was calculated from the relative abundance of species-specific DNA polymorphisms. Gas exchange parameters were measured under well-watered conditions on current-year needle tissues, which were also analyzed for carbon isotope discrimination. Sitka spruce populations had higher gas exchange rates (net photosynthesis, stomatal conductance to water vapor), but lower carbon isotope discrimination values than interior spruce populations. The natural hybrid populations had intermediate values of these parameters consistent with their Si rDNA index. The F(1) population had gas exchange parameters resembling those of Sitka spruce populations, but its carbon isotope discrimination was skewed toward that of interior spruce populations. These results confirmed previous findings that physiological performance of introgressive hybrid spruce populations varied as their DNA constitution changed. PMID- 12651326 TI - Modeling the effect of physiological responses to green pruning on net biomass production of Eucalyptus nitens. AB - Green pruning of Eucalyptus nitens (Deane and Maiden) Maiden increases instantaneous rates of light-saturated CO(2) assimilation (A), and changes patterns of total leaf area and foliage distribution. We investigated the importance of such changes on the rate of recovery of growth following pruning. A simple process-based model was developed to estimate daily net biomass production (G(d)) of three-year-old plantation-grown trees over a 20-month period. The trees had been pruned by removal of 0, 50 or 70% of the length of green crown, equivalent to removal of 0, 55 or 88% of leaf area, respectively, when the plantation verged on canopy closure. Total G(d) was reduced by only 20% immediately following the 50%-pruning treatment, as a result of both the high leaf dark respiration and low A in the portion of the crown removed compared to the top of the crown. Pruning at the time of canopy closure preempted a natural and rapid decline in G(d) of the lower crown. Although leaf area index (L) was approximately 6.0 at the time of pruning, high light interception (95%) occurred with an L of 4.0. The 50%-pruning treatment reduced L to 3.5, but the physiological responses to pruning were sufficient to compensate fully for the reduction in intercepted radiation within 110 days of pruning. The 70%-pruning treatment reduced L to 1.9, and reduced G(d) by 77%, reflecting the removal of branches with high A in the mid and upper crown. Physiological responses to the 70%-pruning treatment were insufficient to increase G(d) to the value of unpruned trees during the study. Model sensitivity analysis showed that increases in A following pruning increased G(d) by 20 and 25% in the 50- and 70%-pruned trees, respectively, 20 months after pruning. Changes in leaf area/foliage distribution had a greater effect on G(d) of 50%-pruned trees (47% increase) than did changes in A. However, the reduction in photosynthetic potential associated with the 70% pruning treatment resulted in only small changes in leaf area/foliage distribution, which consequently had little effect on G(d). The effects of physiological processes occurring within the crown and in response to green pruning on G(d) are discussed with respect to pruning of plantations. PMID- 12651327 TI - Interrelationships among light, photosynthesis and nitrogen in the crown of mature Pinus contorta ssp. latifolia. AB - Scaling leaf-level measurements to estimate carbon gain of entire leaf crowns or canopies requires an understanding of the distribution of photosynthetic capacity and corresponding light microenvironments within a crown. We have compared changes in the photosynthetic light response and nitrogen (N) content (per unit leaf area) of Pinus contorta Dougl. ssp. latifolia Engelm. (lodgepole pine) leaves in relation to their age and light microenvironment. The vertical gradient in integrated daily photosynthetic photon flux density (PPFD) from the upper to the lower crown of lodgepole pine was similar in magnitude to the horizontal gradient in daily PPFD along shoots from young to old leaves. The relationship between light-saturated net photosynthesis (A(max)) and daily PPFD was significant for both young and old leaves. However, old leaves had a lower A(max) than young leaves in a similar daily irradiance regime. For leaves of all ages from throughout the crown, A(max) was linearly related to the estimated daily net carbon gain that leaves could achieve in their natural PPFD environment (estimated A(day)) (r(2) = 0.84, P < 0.001, n = 39), indicating that estimated A(day) may be dominated by carbon fixed when leaves are light-saturated and operating at A(max). Comparison of the PPFD required to achieve A(max) and the PPFD available to the leaves showed that all of the measured leaves (n = 39), regardless of their position in the crown or age, were in light environments that could light-saturate photosynthesis for a similar proportion of the day. For all data pooled, foliar N was weakly correlated with daily PPFD. Analyzing each leaf age class separately showed that foliar N was significantly related to daily PPFD, A(max), and estimated A(day) for the youngest leaves but not for middle aged or old leaves. Therefore, the general theory that foliar N is allocated within a crown according to total daily light availability was supported only for young (1-4 years old) leaves in this study. PMID- 12651328 TI - Ectomycorrhizas and cadmium toxicity in Norway spruce seedlings. AB - We studied the effects of ectomycorrhizal colonization by Laccaria bicolor (Maire) Orton S238 and Paxillus involutus (Batsch) Fr. 533 on cadmium (Cd) toxicity in Norway spruce seedlings (Picea abies (L.) Karst.). Both mycorrhizal and nonmycorrhizal seedlings were exposed to 0 (control), 0.5 or 5 &mgr;M CdSO(4) for 9 weeks in a sand culture system with frequent addition of nutrient solutions. In pure culture, P. involutus and L. bicolor showed similar Cd tolerance. However, in symbiosis, the Cd treatments decreased colonization by L. bicolor but not by P. involutus. Paxillus involutus ameliorated the negative effects of 0.5 &mgr;M Cd on shoot and root growth and chlorophyll content of old needles, whereas L. bicolor did not. Mycorrhizal colonization did not affect Cd concentrations of old needles and roots of seedlings. Despite differences between the ectomycorrhizal fungi in colonization and ability to alleviate Cd toxicity of seedlings, both species reduced Cd concentrations of young needles to a similar degree compared with nonmycorrhizal seedlings. However, in the 0.5 &mgr;M Cd treatment, the Cd content of needles of seedlings colonized by P. involutus was increased, whereas the Cd content of needles of seedlings colonized by L. bicolor was similar to that of needles of nonmycorrhizal seedings. When the amount of Cd translocated to needles was expressed on a root length basis to account for differences in the size of the root systems, the amount of Cd translocated to the needles was similar in seedlings mycorrhizal with P. involutus and in nonmycorrhizal seedlings. All mycorrhizal seedlings were similarly affected by 5 &mgr;M Cd, indicating that the amelioration efficiency of ectomycorrhizal fungi is dependent on the metal concentration to which the roots are exposed. Concentrations of P, K, Ca, Mg and Mn were decreased by 5 &mgr;M Cd to a similar extent in both nonmycorrhizal and mycorrhizal seedlings. In contrast to L. bicolor, P. involutus increased P uptake and altered patterns of root branching. We conclude that mycorrhizas alleviate Cd-induced reductions in growth of Picea abies seedlings. Although the two mycorrhizal fungi examined differed in their ability to alleviate Cd toxicity, these differences were not related to differences in Cd uptake or translocation to the shoot of the mycorrhizal seedlings. We suggest that amelioration of Cd toxicity by P. involutus may be a result of improved P nutrition or changes in root morphology, or both. PMID- 12651329 TI - Shoot and root vulnerability to xylem cavitation in four populations of Douglas fir seedlings. AB - The objectives of this study were to assess the range of genotypic variation in the vulnerability of the shoot and root xylem of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings to water-stress-induced cavitation, and to assess the trade-off between vulnerability to cavitation and conductivity per unit of stem cross-sectional area (k(s)), both within a species and within an individual tree. Douglas-fir occupies a broad range of environments and exhibits considerable genetic variation for growth, morphology, and drought hardiness. We chose two populations from each of two varieties (the coastal var. menziesii and the interior var. glauca) to represent environmental extremes of the species. Vulnerability curves were constructed for shoots and roots by plotting the percentage loss in conductivity versus water potential. Vulnerability in shoot and root xylem varied genetically with source climate. Stem xylem differed in vulnerability to cavitation between populations; the most mesic population, coastal wet (CW), was the most susceptible of the four populations. In the roots, the most vulnerable population was again CW; the interior wet (IW) population was moderately susceptible compared with the two dry populations, coastal dry (CD) and interior dry (ID). Root xylem was more susceptible to cavitation than stem xylem and had significantly greater k(s). The trade-off between vulnerability to cavitation and k(s), however, was not evident across populations. The most vulnerable population (CW) had a shoot k(s) of 0.534 +/- 0.067 &mgr;mol m(-2) s( 1) MPa(-1), compared with 0.734 +/- 0.067 &mgr;mol m(-2) s(-1) MPa(-1) for the less vulnerable CD stems. In the roots, IW was more vulnerable than ID, but had the same k(s). PMID- 12651330 TI - Differences in chemical composition relative to functional differentiation between petioles and laminas of Fraxinus excelsior. AB - Differences in structural and nonstructural carbohydrates, lignin and chlorophyll, and Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase) activity between petioles and leaflets were studied along a canopy light gradient in Fraxinus excelsior L., which has pinnate compound leaves and up to 20% of foliar biomass invested in petioles. Long-term light conditions at the sampling locations were characterized by values of seasonal mean integrated quantum flux density (Q(int), mol m(-2) day(-1)) estimated by combining data from hemispherical photographs at the sampling locations with measurements of global solar radiation above the canopy during the growing season. The contribution of petioles to leaf carbon assimilation was disproportionally lower than that of leaf laminas. Though the light relationships of assimilative compounds-foliar chlorophyll concentration increasing with decreasing Q(int) to improve leaf absorptance, foliar N concentration and Rubisco activity being relatively constant along the light gradient-were similar for both petioles and leaflets, petiole nitrogen and chlorophyll concentrations were only 30% and 10%, respectively, of those of leaflets. Nonstructural carbohydrate concentration was about 20% higher in petioles than in leaf laminas, indicating that petioles also serve as storage tissues for photosynthates. Relationships between foliar structural carbon components and irradiance-increasing lignin (L) and decreasing structural polysaccharide (SP) concentrations with increasing Q(int)-were qualitatively similar for petioles and leaflets. However, petioles had lower L, but higher SP and total investment in structural compounds (L + SP) than leaflets. Greater lignification at high irradiances in leaflets than in petioles was attributed to greater water stresses at high light, and to more variable water contents of actively transpiring leaflets. Low lignin concentration in combination with high osmotically active carbohydrate concentrations in petioles suggest that turgor plays an important role in the mechanical properties of petioles. As a result of lower lignin and protein concentrations, the glucose cost of petiole construction (g glucose per g dry mass) was about 5% lower than that of leaf laminas. PMID- 12651331 TI - Variation in net photosynthesis, stomatal characteristics, leaf area and whole plant phytomass production among ten provenances of neem (Azadirachta indica). AB - Variation in net photosynthesis, CO(2) exchange parameters, stomatal characteristics, leaf area and seedling dry weight were investigated among 10 provenances of neem (Azadirachta indica A. Juss.). Significant provenance variation was established for net photosynthesis (8.14 to 15.13 &mgr;mol m(-2) s( 1)), stomatal conductance (0.37 to 0.59 mol m(-2) s(-1)), stomatal density (145 to 204 mm(-2)), and total guard cell length (2681 to 3873 &mgr;m). Net photosynthesis was positively correlated with whole-plant dry weight and leaf area. Stomatal density was positively correlated with net photosynthesis, whole plant dry weight, and leaf area. Total guard cell length was positively correlated with all of these traits. Information on six traits was used in a cluster analysis to construct a dendrogram to assess phenetic relationships among the provenances. With a few exceptions, the dendrogram revealed three major clusters grouped according to rainfall distribution. The study indicated that whole-plant phytomass production of neem seedlings was associated with photosynthesis and stomatal characteristics during the early stages of growth. PMID- 12651332 TI - The effect of aqueous transport of CO(2) in xylem sap on gas exchange in woody plants. AB - The influence of CO(2) transported in the transpiration stream on measurements of leaf photosynthesis and stem respiration was investigated. Measurements were made on trees in a temperate forest in Scotland and in a tropical rain forest in Cameroon, and on shrubs in the Sahelian zone in Niger. A chamber was designed to measure the CO(2) partial pressure in the gas phase within the woody stems of trees. High CO(2) partial pressures were found, ranging from 3000 to 9200 Pa. Henry's Law was used to estimate the CO(2) concentration of xylem sap, assuming that it was in equilibrium with the measured gas phase partial pressures. The transport of CO(2) in the xylem sap was calculated by multiplying sap CO(2) concentration by transpiration rate. The magnitude of aqueous transport in the studied species ranged from 0.03 to 0.35 &mgr;mol CO(2) m(-2) s(-1), representing 0.5 to 7.1% of typical leaf photosynthetic rates. These values strongly depend on sap pH. To examine the influence of aqueous transport of CO(2) on stem gas exchange, we made simultaneous measurements of stem CO(2) efflux and sap flow on the same stem. After removing the effect of temperature, stem CO(2) efflux was positively related to sap flow. The apparent effect on measurements of stem respiration was up to 0.7 &mgr;mol m(-2) s(-1), representing ~12% of peak stem respiration rates. PMID- 12651333 TI - Biomass and nutrient allocation in Douglas-fir and amabilis fir seedlings: influence of growth rate and temperature. AB - Allocation of biomass and nutrients to current-year and one-year-old shoots and roots of two-year-old conifer seedlings with differing rates of growth was studied. Differences in growth rate were achieved by selecting fast- and slow growing populations of the relatively fast- and slow-growing conifer species, Pseudotsuga menziesii (Mirb.) Franco and Abies amabilis Dougl. ex Loud, respectively. Environmentally controlled differences in growth rate were induced by placing half of the seedlings in a 10 degrees C growth chamber and half in a 20 degrees C growth chamber in their second growing season. Seedling samples were harvested in May, before the temperature treatment, and in July and November of the second growing season, and biomass and nutrient concentrations of current year and one-year-old shoots and roots were determined. Seedling biomass and nutrient allocation differed among the high-growth treatments. Seedlings exhibiting high growth in response to the 20 degrees C treatment and faster growing populations within species both showed increased allocation to new shoots, whereas seedlings of the fast-growing species showed greater allocation to old shoots than to new shoots. Increased growth increased nutrient uptake, but nutrient concentration decreased with growth rate as a result of dilution, so that faster-growing seedlings had greater nutrient-use efficiency than slower growing seedlings. Retranslocation of P and K was seen in the second year only in slow-growing populations at 10 degrees C. Nutrient concentrations of one-year-old plant parts decreased in the second year, indicating new growth was a stronger sink for nutrients than second-year growth. PMID- 12651334 TI - Freezing cycles enhance winter injury in Picea rubens. AB - We examined changes in chlorophyll absorbency in red spruce (Picea rubens Sarg.) foliage in response to simulated freezing cycles. Current-year branch tips were collected from 16 trees on January 8, January 20, February 8 and February 26, 1996. Tissue was subjected to freezing cycle treatments with a minimum of -35 degrees C and a maximum of 3 degrees C for a one-cycle treatment, and -9, -6, -3, 0 or 3 degrees C for four-cycle treatments. Samples were frozen at a rate of 5 degrees C h(-1), and warmed at 12 to 15 degrees C h(-1). Controls were held at -9 degrees C. Temperatures during the three-day periods preceding each sample date averaged -18, 4.7, -9.6 and 3.7 degrees C, respectively. On January 8, treated trees showed no significant (P > 0.1) increase in the breakdown of chlorophyll, as measured by the ratio of chlorophyll a absorbency (435 nm) to phaeophytin a absorbency (415 nm), compared with control branch tips. On later sampling dates, seven trees consistently exhibited needle reddening and nine exhibited few symptoms (< 10% of total needle surface reddened) after four-cycle treatments. On February 26, chlorophyll degradation in trees with needle reddening differed (P < 0.05) from the control by 26, 26, 16, 14 and 15% for the 3, 0, -3, -6 and -9 degrees C maxima, respectively. No detectable chlorophyll degradation occurred after a one-cycle treatment in any trees on any sampling date. Freezing cycles with sub-zero maxima and a -35 degrees C minimum enhanced winter injury in red spruce after a midwinter thaw had rendered the trees susceptible to freezing damage. PMID- 12651335 TI - Lessons learned while extending physiological principles from growth chambers to satellite studies. AB - Over the last three decades, physiological principles established in laboratory studies have been applied to systems at progressively larger scales and are now firmly merged into the fields of ecology, ecosystem modeling, forest protection, and global change research. To expand the vision of any field requires that scientists from different disciplines build a bridge across the chasm that normally exists between the knowledge bases and perspectives of different fields. Bridges are built most quickly when representatives of different disciplines see the possibility of mutual advantage in collaboration and seek to quickly demonstrate that potential. Usually, however, the process is laborious because approaches and techniques must be modified to address problems at a different level of integration. Successful bridge builders have, almost without exception, established credibility in their own field and have then identified a kindred spirit with similar credentials in another. They usually establish a pilot study that involves apprentices as well as established scientists. If the approach is successful, the younger members of the team often take the lead in further advancements. Managers of large centralized programs should foster interdisciplinary exchange, particularly at times when advancement in one field languishes. To expand collaboration, it is often necessary for scientists to seek common properties that simplify relations across a wide range of biological and physical conditions. This integrative perspective is essential and is fostered by participating in cross-disciplinary workshops and conferences and by reading outside one's field. PMID- 12651336 TI - A review of whole-plant water use studies in tree. AB - Weighing lysimeters, large-tree potometers, ventilated chambers, radioisotopes, stable isotopes and an array of heat balance/heat dissipation methods have been used to provide quantitative estimates of whole-tree water use. A survey of 52 studies conducted since 1970 indicated that rates of water use ranged from 10 kg day(-1) for trees in a 32-year-old plantation of Quercus petraea L. ex Liebl. in eastern France to 1,180 kg day(-1) for an overstory Euperua purpurea Bth. tree growing in the Amazonian rainforest. The studies included in this survey reported whole-tree estimates of water use for 67 species in over 35 genera. Almost 90% of the observations indicated maximum rates of daily water use between 10 and 200 kg day(-1) for trees that averaged 21 m in height. The thermal techniques that made many of these estimates possible have gained widespread acceptance, and energy balance, heat dissipation and heat-pulse systems are now routinely used with leaf level measurements to investigate the relative importance of stomatal and boundary layer conductances in controlling canopy transpiration, whole-tree hydraulic conductance, coordinated control of whole-plant water transport, movement of water to and from sapwood storage, and whole-plant vulnerability of water transport to xylem cavitation. Techniques for estimating whole-tree water use complement existing approaches to calculating catchment water balance and provide the forest hydrologist with another tool for managing water resources. Energy-balance, heat dissipation and heat-pulse methods can be used to compare transpiration in different parts of a watershed or between adjacent trees, or to assess the contribution of transpiration from overstory and understory trees. Such studies often require that rates of water use be extrapolated from individual trees to that of stands and plantations. The ultimate success of this extrapolation depends in part on whether data covering short time sequences can be applied to longer periods of time. We conclude that techniques for estimating whole-tree water use have provided valuable tools for conducting basic and applied research. Future studies that emphasize the use of these techniques by both tree physiologists and forest hydrologists should be encouraged. PMID- 12651337 TI - Carbon and oxygen isotope ratios of ecosystem respiration along an Oregon conifer transect: preliminary observations based on small-flask sampling. AB - Isotope ratio analyses of atmospheric CO(2) at natural abundance have significant potential for contributing to our understanding of photosynthetic and respiration processes in forest ecosystems. Recent advances in isotope ratio mass spectrometry allow for rapid, on-line analysis of small volumes of CO(2) in air, and open new research opportunities at the ecophysiological, whole-organism, and atmospheric levels. Among the immediate applications are the carbon and oxygen isotope ratio analyses of carbon dioxide in atmospheric air. Routine analysis of carbon dioxide in air volumes of approximately 50-300 &mgr;l is accomplished by linking a commercially available, trace gas condenser and gas chromatograph to an isotope ratio mass spectrometer operated in continuous-flow mode. Samples collected in the field are stored in either gas-tight syringes or 100-ml flasks. The small sample volume required makes it possible to subsample the air in flasks for CO(2) and then to sample the remaining air volume for the analysis of the isotopic composition of either methane or nitrous oxide. Reliable delta(13)C and delta(18)O values can be obtained from samples collected and stored for 1-3 days. Longer-term storage, on the order of weeks, is possible for delta(13)C measurements without drift in the isotope ratio signal, and should also be possible for delta(18)O measurements. When linked with an infrared gas analyzer, pump and flask sampling system, it is feasible to sample CO(2) extensively in remote forest locations. The air-sampling system was used to measure the isotope ratios of atmospheric CO(2) and to conduct a regression analysis of the relationship between these two parameters. From the regression, we calculated the delta(13)C of ecosystem respiration of four coniferous ecosystems along a precipitation gradient in central Oregon. The ecosystems along the coast-to interior Oregon (OTTER) gradient are dominated by spruce-hemlock forests at the wet, coastal sites (> 200 cm precipitation annually) to juniper woodlands (20 cm precipitation) at the interior, dry end of the transect. The delta(13)C values of ecosystem respiration along this transect differed by only 1.3 per thousand (range of -25.2 to -23.9 per thousand ) during August at the peak of the summer drought. Following autumn rains in September, the delta(13)C of ecosystem respiration in the four stands decreased; overall the difference in the carbon isotope ratio of ecosystem respiration among sites increased to 3.9 per thousand (-26.8 to -22.9 per thousand ). PMID- 12651338 TI - Freezing behaviors in leaf buds of cold-hardy conifers visualized by NMR microscopy. AB - A calibration curve was established to convert plant area index of Eucalyptus nitens (Deane and Maiden) Maiden, assessed with a Li-Cor LAI-2000, to leaf area index, LAI. Based on a comparison of this calibration curve with existing calibration curves for other species, we concluded that a generic calibration curve may be applicable for the assessment of LAI in eucalypt plantations. The Li Cor LAI-2000 measurements were used to correlate the equilibrium LAI of E. nitens plantations with mean annual temperature. These and other data were then combined to develop relationships between LAI in both E. nitens and E. globulus Labill. plantations and mean annual temperature and water stress. In plantations of both species, LAI declined linearly with water stress. However, marked differences in the effect of suboptimal growth temperatures on LAI were observed between species: on cold sites, LAI of E. nitens was markedly higher than LAI of E. globulus. A simple analytic model of net primary production (NPP) was developed. In this model, increasing LAI increased light interception and hence dry matter production, but simultaneously increased canopy respiration. Consequently, for a given light utilization coefficient (epsilon), there was a value of LAI that maximized NPP. The model was parameterized for E. globulus and used to investigate the influences of water stress and mean annual temperature on LAI through their effects on epsilon. The model indicated that the value of LAI that was predicted to maximize NPP under various water and temperature stress regimes was similar to the value of LAI observed in the field under similar conditions only if leaf longevity was linked to water and temperature stress. PMID- 12651339 TI - Does leaf water efficiency vary among eucalypts in water-limited environments? AB - There is a need to generalize water use behavior of eucalypts to facilitate bioengineering and landscape remediation programs in a wide range of Australian environments. A critical question can be stated as a null hypothesis: tree water use per unit leaf area (leaf efficiency) is independent of eucalypt species. This is implicitly equivalent to the hydrological equilibrium hypothesis that states that leaf area is a function of climate, at least in cases where transpiration and growth are limited by soil water. Failure to reject this null hypothesis simplifies (a) the selection of tree species for water balance management, (b) the generation of regional-scale expectations of leaf area index, and (c) the estimation (monitoring) of the effectiveness of plantations in controlling site water balance. The hypothesis was tested with tree water use data collected in natural multi-species stands across Australia, including sites in the wet-dry season tropical woodlands of the Northern Territory, the Mediterranean climate forests of Western Australia, and a woodland system in southern New South Wales receiving an even distribution of rainfall throughout the year. We also tested the hypothesis in a multi-species tree plantation growing on a saline gradient. In each case, we could not reject the hypothesis of constant leaf efficiency among eucalypts. In every case there was a common, strong, linear relationship among tree leaf area and mean daily water use by all tree species in a sample. Single factor (species) analysis of variance did not detect significant differences between leaf water efficiencies of species. For the jarrah forest (Eucalyptus marginata J. Donn ex Sm., E. calophylla R. Br. ex Lindl.), the null hypothesis held in both spring (wet) and autumn (dry) conditions. The null hypothesis held in the mixed species woodland of New South Wales (E. macrorhynca F.J. Muell. ex Benth., E. blakelyi Maiden., E. polyanthemos Schauer.) under summer and autumn conditions, and across five species in the wet-dry tropical woodland (E. miniata A. Cunn. ex Schauer, E. tetrodonta F.J. Muell., E. porecta S.T. Blake, Erythrophleum chlorostachys F.J. Muell., and Terminalia ferdinandiana Exell.). The null hypothesis also held for a plantation of E. occidentalis Endl. and provenances of E. camaldulensis Dehnh. growing on a shallow saline gradient; i.e., leaf water efficiency remained constant across species and varieties despite obvious effects of salinity on the size of individual canopies. We conclude that there is little evidence for rejecting the hypothesis that leaf efficiency does not vary significantly among sympatric eucalypt species in rainfall-limited (soil-water-limited) systems. These findings open the way for useful bioengineering generalities about the hydrological role of trees in the Australian landscape. PMID- 12651340 TI - A cost-benefit analysis of leaves of four Australian savanna species. AB - We conducted a cost-benefit analysis of the contrasting phenologies of two evergreen and two deciduous species of the savannas of north Australia. Stomatal conductance, rates of light-saturated assimilation (A(max)) and dark respiration were measured for six leaves from each of five or six trees. These leaves were then analyzed for total nitrogen, ether-soluble lipids, ash content, and heat of combustion. Construction and maintenance costs, nitrogen-use efficiencies and instantaneous transpiration efficiencies were then calculated from these data. Evergreen species had significantly lower specific leaf area, leaf nitrogen and leaf ash content than deciduous species. Evergreen species also had significantly higher heat of combustion and lipid content of crude extracts than deciduous species. Light-saturated assimilation rates were higher in evergreen species on a leaf area basis, but were higher in deciduous species on a leaf dry weight basis. In both evergreen and deciduous species, A(max) and total Kjeldahl nitrogen were linearly related. Similarly, nitrogen-use efficiency did not differ among species. Leaf construction costs were significantly higher for evergreen species than for deciduous species, but maintenance costs did not differ among species. Evergreen species had a higher cost:benefit ratio than deciduous species but because of their longer-lived leaves, the payback interval was longer in evergreen species than in deciduous species. These results support the hypotheses that: (1) longer-lived leaves are more expensive to construct than shorter-lived leaves, and (2) there is a higher investment of nitrogen into short-lived leaves to support a higher A(max) over a shorter payback interval. We conclude that deciduous and evergreen species partition resources both temporally and spatially, thereby reducing interspecies competition. PMID- 12651341 TI - Transpiration and canopy conductance in a eucalypt plantation using shallow saline groundwater. AB - Tree water use and canopy conductance were monitored in a 20-year-old Eucalyptus grandis W. Hill ex Maiden and Eucalyptus camaldulensis Dehnh. plantation shown to be using shallow saline groundwater. Annual water use, obtained from two years of heat pulse measurements of daily water use, was approximately 300 mm for both species with a winter minimum and a weak maximum in spring. Canopy conductance obtained from the ratio of daily water use to daily mean vapor pressure deficit (VPD) was low during summer, but rose to high values in winter. Diurnal variation in stomatal conductance was recorded in late summer and spring. Stomatal conductance increased with radiation but showed no evidence of a relationship with VPD. Canopy conductance was generally less than half the observed stomatal conductance on a leaf area basis, and usually declined steadily through the day, showing a strong inverse relationship with VPD. For both species, a decoupling coefficient (Omega) of 0.1 to 0.3 was obtained from canopy conductance and climate observations, and a higher value of Omega was derived from canopy and stomatal conductances. The difference in Omega values is interpreted as demonstrating the inclusion of a soil to leaf conductance within the canopy conductance estimates. Soil to leaf conductance is smaller than both stomatal and aerodynamic conductances, and effectively limits water use by the plantation. The observed variation in soil to leaf conductance is consistent with resistance to water movement to the roots increasing as the soil in the vicinity of the active roots dries as a result of water uptake during the day. Plantations may be useful for transpiring shallow ground-water to control rising water tables and salinity, but their effectiveness as water users will be reduced as the water table is drawn down in soils of low hydraulic conductivity. Plantations irrigated with pumped groundwater or drainage water may provide a more efficient disposal system where these water sources are available. PMID- 12651342 TI - Whole-tree transpiration and water-use partitioning between Eucalyptus nitens and Acacia dealbata weeds in a short-rotation plantation in northeastern Tasmania. AB - Whole-tree water use in 4- and 8-year-old plantations of Eucalyptus nitens Deane and Maiden (ex Maiden) in the presence and absence of Acacia dealbata Link. weeds was estimated by the heat pulse velocity technique during a six-week summer period. Maximum sap velocities were recorded between 5 and 15 mm under the cambium for both eucalypt and acacia trees, and marked radial and axial variations in sap velocity were observed. The latter source of variation was most pronounced in mixed stands where crowns were asymmetrical. Mean daily sap flux ranged from 1.4 to 103.6 l day(-1) for eucalypts and from < 0.1 to 8.4 l day(-1) for acacias. Stem diameter explained 98% of the variation in sapwood area for E. nitens and 89% for A. dealbata, and was determined to be a suitable parameter for scaling water use from the tree to stand level. Plot transpiration varied from 1.4 to 2.8 mm day(-1) in mixed 8-year-old plots and was 0.85 mm day(-1) in a mixed 4-year-old plot. The degree of A. dealbata infestation was associated with absolute plot water use and regression models predicted that, in the absence of acacia competition, plot water use for the 8-year-old stand would approach 5-6 mm day(-1) during the growing season. PMID- 12651343 TI - Response of effluent-irrigated Eucalyptus grandis and Pinus radiata to salinity and vapor pressure deficits. AB - Effects of high vapor pressure deficit (VPD) and soil salinity on growth and physiology of Pinus radiata D. Don and Eucalyptus grandis Hill ex Maiden were studied in a five-year-old plantation irrigated with salt-enhanced effluent (2.2 dS m(-1)) or freshwater (0.2 dS m(-1)) for 14 weeks during spring and summer. Salt was then rapidly leached by over-irrigation with low-salinity effluent. Soil water and salinity, tree water stress, sap flux, substrate carbon conversion efficiency, foliage and stem growth, and foliar cations and chloride were monitored throughout the study. An average of 9 and 1 Mg ha(-1) of salt with an average hydraulic load of 660 and 780 mm was applied to the salt and control plots, respectively. Maximum soil salinity in the root zone was 5.8 and 6.8 dS m( 1) in the eucalypt and pine plots, respectively. Predawn water potential was more than twice as sensitive to increasing salinity in E. grandis as in P. radiata. The salt treatment reduced rates of leaf and stem growth of the eucalypts by 60 to 70% but had no effect on leaf and stem growth of the pines. In the eucalypts, salinity decreased mean leaf area by 26% and increased specific leaf area by 12% compared with control values, indicating less biomass per unit leaf area in the salt treatment. Salinity had no effects on these two parameters in pine. The salt treatment significantly increased mean foliar concentrations of Na and Cl in both species, and of K in the pines. Foliar Na concentration was 6-10 times higher in the eucalypts than in the pines. Lowered water potential and increased Na concentration in the eucalypts in response to salinity resulted in about a 50% reduction in the efficiency of conversion of carbon into biomass; however, three weeks after leaching the salt, there was no significant difference in efficiency of conversion of carbon into biomass between the treatments. Salinity had no effect on water use by eucalypts, but caused a nonsignificant decrease (7%) in water use by pines. As evaporative demand increased, crop factor (transpiration divided by pan evaporation) declined by up to 50 and 60% in the pines and eucalypts, respectively. We conclude that stomatal response to high VPD, not soil salinity, accounts for most of the reduction in summertime water use. PMID- 12651344 TI - Transpiration rates and canopy conductance of Pinus radiata growing with different pasture understories in agroforestry systems. AB - We measured tree transpiration and canopy conductance in Pinus radiata D. Don at two low rainfall sites of differing soil fertility in Canterbury, New Zealand. At the more fertile Lincoln site, we also assessed the effects of two common pasture grasses on tree transpiration and canopy conductance. At the less fertile Eyrewell Forest site, the effect of no understory, and the effects of irrigation in combination with mixtures of grass or legume species were determined. Tree xylem sap flux (F(d)') was measured by the heat pulse method. Total canopy conductance to diffusion of water vapor (G(t)) was calculated by inverting a simplified Penman-Monteith model. The different treatment effects were modeled by the simple decaying exponential relationship G(t) = G(tmax)e((-bD)), where D = air saturation deficit. At the Lincoln site, trees with an understory of cocksfoot had lower F(d)' and G(tmax) than trees with an understory of ryegrass, although the sensitivity of G(t) to increasing D (i.e., the value of b) did not differ between treatments. At the Eyrewell site, irrigation only increased F(d)' in the absence of an understory, whereas the presence of understory vegetation, or lack of irrigation, or both, significantly reduced G(tmax) and increased b. We conclude that the selection of understory species is critical in designing successful agroforestry systems for low rainfall areas. PMID- 12651345 TI - Vulnerability to drought-induced embolism of Bornean heath and dipterocarp forest trees. AB - Occasional droughts may be important in controlling the distribution and structure of forest types in relatively aseasonal north Borneo. The low water retention capacity of the coarse, sandy soils on which tropical heath forest occurs may cause drought to develop more quickly and severely than on the finer textured soils of nearby dipterocarp forest. Resistance to drought-induced embolism is considered an important component of drought tolerance. We constructed embolism vulnerability curves relating loss in hydraulic conductivity to xylem tension by the air-injection method for understory trees of 14 species from both tropical heath and mixed dipterocarp forests in Brunei Darussalam. There was no significant difference (Mann-Whitney U-test, P = 0.11) between forest types in the xylem tension at which 50% loss of hydraulic conductivity occurred. Most species from both forest types were highly vulnerable to embolism compared with species from seasonal tropical forests. We speculate that other mechanisms, such as stomatal control to prevent development of embolism-inducing xylem tensions, are more cost-effective adaptations against occasional drought, but that the attendant reduction in productivity and competitive ability places a greater premium on resistance to embolism when drought is annual and predictable. PMID- 12651346 TI - A mathematical and statistical analysis of the curves illustrating vulnerability of xylem to cavitation. AB - When vulnerability curves are used to assess the susceptibility of plants to drought, the water potential corresponding to 50% loss of conductivity is frequently used as a measure of susceptibility. However, this value does not distinguish between different patterns of conductivity loss, such as a rapid loss over a narrow water potential range versus a gradual decline in conductivity. We have applied an exponential sigmoidal equation to vulnerability curves obtained from four closely related Eucalyptus clones differing in drought tolerance. The coefficients of the equation were evaluated and statictically compared among the clones. If the air-seedling hypothesis of cavitation is accepted, these coefficients have biological relevance. One of the coefficients describes the position of the curve on the water potential axis and is equivalent to the water potential corresponding to 50% loss of conductivity. This coefficient could reflect the size of the largest pit pore per xylem vessel. Another coefficient is related to the slope of the conductivity loss, and could reflect the range in maximum pit pore size per vessel. PMID- 12651347 TI - Relationship between growth and xylem hydraulic characteristics of clones of Eucalyptus spp. at contrasting sites. AB - To assess relationships between hydraulic characteristics and growth efficiency (expressed as trunk volume increment per unit leaf area per year), we measured xylem specific conductivity, leaf specific conductivity and the vulnerability of xylem to cavitation in branches of 7-8-year-old trees of four closely related Eucalyptus clones growing in plantations on mesic and xeric sites. Growth was influenced more by site than by clone. Hydraulic conductivities were higher in trees on mesic sites than in trees on xeric sites, but within a site, conductivity did not differ among clones. Huber values (functional xylem cross sectional area:leaf area) were similar across clones and between sites. Vulnerability of xylem to cavitation differed among clones but not between sites. We conclude that, within these genetically related clones, hydraulic conductivity is determined predominantly by environmental conditions, whereas vulnerability to cavitation is more closely associated with genotype. The clone most vulnerable to cavitation showed considerable aboveground die-back during a severe drought and there was a positive correlation between growth efficiency and maximum xylem specific conductivity. PMID- 12651348 TI - Fine roots dynamics in a Mediterranean forest: effects of drought and stem density. AB - Summer drought is a characteristic trait of Mediterranean climates and can limit primary production. The holm oak (Quercus ilex L.) forest at the Prades Experimental Complex of Catchments (PECC, NE Spain) was managed by clearcutting until the 1950s. As a result of this practice, the forest has a high density (13000 stems ha(-1)) that is causing growth stagnation and intense competition for water. A thinning experiment was conducted to release resources, reduce stem competition and improve forest productivity. Minirhizotrons were installed in two plots in each of the thinned and control stands and fine root dynamics quantified to a soil depth of 60 cm over a 2-year period (June 1994-May 1996). Soil water content was higher and soil water penetrated deeper in the soil (30-50 cm) in the thinned plots than in the control plots. In the control plots, soil temperature did not vary substantially with depth (mean = 11.65 degrees C), whereas soil temperature was higher in shallow horizons compared with deep horizons in the thinned plots. Mean soil temperature was one degree higher in the thinned plots than in the control plots (12.9 versus 11.9 degrees C). There were always more roots in the top 20 cm of soil in thinned plots than in control plots. In the thinned plots, more than 50% of root density (root no. cm(-2)) was located in the top 20 cm of soil, whereas root density never reached 50% in the 0-20 cm soil layer of the control plots. Roots longevity varied from 35 to 471 days, and there were significant differences between soil depths. Root longevity was greater in the control plots than in the thinned plots. PMID- 12651349 TI - Photosynthesis and photoprotection in Quercus ilex resprouts after fire. AB - Plants that resprout after fires often have higher rates of photosynthesis than before a fire. To elucidate the mechanism of this response, we studied gas exchange and chlorophyll fluorescence in Quercus ilex L. plants growing on control (unburned) sites and on sites that had been burned the preceding summer. In early July, photosynthetic rates and stomatal conductance were similar in plants on unburned and burned plots, and in young and old foliage within unburned plots. At this time, photochemical efficiency of photosystem II (PSII), nonphotochemical quenching of chlorophyll fluorescence (NPQ), and the de epoxidation of violaxanthin to zeaxanthin were also similar among leaves of different ages and treatments. In late July, photosynthetic rates and stomatal conductances were much greater in resprouts on the burned areas than in unburned plants. From early to late July, unburned plants showed an increase in NPQ and the de-epoxidation of violaxanthin to zeaxanthin, indicating increased photoprotection as a result of enhanced nonradiative dissipation of excess light energy. Plants on the burned plots did not show these changes. Leaves of all ages and treatments showed no substantial reduction in potential quantum yield of PSII (F(v)/F(m)) at midday or predawn, indicating that there was little or no photoinhibition. Leaf nitrogen and soluble protein contents varied with leaf age during July, but did not vary between treatments. We conclude that the primary effect of burning is an increase in water availability to resprouting plants that eliminates the need for photoprotection, at least in the short term. The decrease in photosynthetic rates of unburned leaves in late July was the result of reduced stomatal conductance. We suggest that lowered stomatal conductance is the primary limiting factor in Q. ilex leaves, governing the regulation of carboxylation activity and energy dissipation processes. PMID- 12651350 TI - Passive nighttime warming facility for forest ecosystem research. AB - A nighttime warming experiment is proposed. Over the last four decades a significant rise in nighttime minimum temperature has been determined from analysis of meteorological records from a global distribution of locations. The experiment involves nighttime deployment of infrared (IR) reflecting curtains around four sides of a forest canopy and across the top of the forest to mimic the top-down warming effect of cloud cover. The curtains are deployed with cable and pulley systems mounted on a tower and scaffolding structure built around the selected forest site. The trunk space is not enclosed except as an optional manipulation. The curtains reflect long-wave radiation emitted from the forest and ground back into the forest warming the trees, litter, and soil. Excellent infrared reflection can be obtained with commercially available fabrics that have aluminum foil bonded to one side. A canopy warming of 3 to 5 degrees C is expected on cloudless nights, and on cloudy nights, a warming of 1 to 3 degrees C is anticipated relative to a control plot. The curtains are withdrawn by computer control during the day and also at night during periods with precipitation or excessive wind. Examples of hypothesized ecosystem responses to nighttime warming include: (1) increase in tree maintenance respiration (decreasing carbon reserves and ultimately tree growth), (2) increase in the length of the growing season (increasing growth), (3) increase in soil respiration, (4) increase in litter decomposition, (5) increase in mineralization of N and other nutrients from soil organic matter, (6) increase in nutrient uptake (increasing growth), and (7) increase in N immobilization in litter. Hypothesis 1 has the opposite consequence for tree growth to Hypotheses 2 and 6, and thus opposite consequences for the feedback regulation that vegetation has on net greenhouse gas releases to the atmosphere. If Hypothesis 1 is dominant, warming could lead to more warming from the additional CO(2) emissions. Site-specific meteorological, ecophysiological, and phenological measurements are obtained in the warming treatment and in a carefully selected control plot to investigate site-specific hypotheses. Measurements made on both plots for a baseline period and during the period of curtain deployment provide data to test the hypotheses statistically by the "before-after-control-impact" method applicable to unreplicated experiments. The enclosure has a modular design that can be adapted and combined with other forest scale manipulation experiments such as free air CO(2) enrichment and throughfall displacement. PMID- 12651351 TI - Water use by forests, limits and controls. AB - Based on a review of several studies that have been carried out to determine the water use of forests in relation to other crops in different regions of the world, it is shown that the principal controls on evaporation from forests and shorter crops vary markedly between the temperate and tropical regions and between the wet and dry zones of these regions. Although there are detailed physical and physiological models available that allow calculation of forest water use, these models are not always readily applicable. It is proposed that a knowledge of the limits on evaporation can be used to devise models of varying complexity for estimating water use of forests in different regions and for predicting differences in water use between forests and shorter crops. Limits on evaporation may be related to radiation, advection, tree physiology, soil water, tree size or drop size. Examples are given of the use of models based on the limits concept for solving forest related water resource problems in Malawi and the U.K. PMID- 12651352 TI - Regulation of stomatal conductance and transpiration in forest canopies. AB - Processes regulating stomatal conductance, g(s), and transpiration, E, from forest canopies are reviewed. The first section deals with the response of g(s) to environmental variables. Phenomenological models have been used to interpret field data and predict diurnal and seasonal variability in g(s), but models that couple stomatal conductance to photosynthesis at the leaf scale are now being used more widely. The vertical distribution of foliar nitrogen concentration is helpful for scaling these processes from leaves to canopies, and the analysis of data from many studies has led to the emergence of simplified, general relationships for estimating evaporation and carbon uptake by forests at stand and regional scales. Evidence for the regulation of stomatal conductance by hydraulic and chemical signals is presented in the second section. Rapid and reversible changes in g(s) following a perturbation to the water potential gradient in the flow pathway suggest that stomata respond directly to hydrostatic signals. Other evidence supports the contention that signals are transmitted by abscisic acid (ABA), possibly originating in the roots. For large woody plants, the short-term responses of stomata are probably brought about by hydraulic signals that affect g(s) by triggering the release of ABA in the leaves. Tardieu and Davies (1993) developed an interactive model that incorporates hydraulic and chemical effects to describe the response of stomata to soil drying and evaporative demand. In the third section, evidence is presented that short-term changes in g(s) are linked closely to the hydraulic properties of the conducting system to minimize loss of hydraulic conductivity through xylem by cavitation. Examples of homeostatic mechanisms that operate to ensure the long-term balance between evaporative demand and the potential hydraulic conductivity of trees growing in different environments are described. Two hypotheses are examined: (1) height growth in trees is limited by the capacity of the conducting system; and (2) the decline in productivity with stand age is attributable to a decrease in conductivity. PMID- 12651353 TI - Drought tolerance, xylem sap abscisic acid and stomatal conductance during soil drying: a comparison of young plants of four temperate deciduous angiosperms. AB - Patterns of water relations, xylem sap abscisic acid (ABA) concentration ([ABA]) and stomatal aperture were compared in drought-sensitive black walnut (Juglans nigra L.) and black willow (Salix nigra Marsh.), less drought-sensitive sugar maple (Acer saccharum Marsh.) and drought-tolerant white oak (Quercus alba L.). Strong correlations among reduction in predawn water potential, increase in xylem sap [ABA] and stomatal closure were observed in all species. Stomatal response was more highly correlated with xylem [ABA] than with ABA flux. Xylem sap pH and ion concentrations appeared not to play a major role in the stomatal response of these species. Stomata were more sensitive to relative changes in [ABA] in drought-sensitive black walnut and black willow than in sugar maple and white oak. In the early stages of drought, increased [ABA] in the xylem sap of black walnut and black willow was probably of root origin and provided a signal to the shoot of the water status of the roots. In sugar maple and white oak, leaf water potential declined with the onset of stomatal closure, so that stomatal closure also may have occurred in response to the change in leaf water potential. PMID- 12651354 TI - Drought tolerance, xylem sap abscisic acid and stomatal conductance during soil drying: a comparison of canopy trees of three temperate deciduous angiosperms. AB - Patterns of water relations, xylem sap abscisic acid concentration ([ABA]) and stomatal aperture were characterized and compared in drought-sensitive black walnut (Juglans nigra L.), less drought-sensitive sugar maple (Acer saccharum Marsh.) and drought-tolerant white oak (Quercus alba L.) trees co-occurring in a second-growth forest in Missouri, USA. There were strong correlations among reduction in predawn leaf water potential, increased xylem sap [ABA] and stomatal closure in all species. Stomatal conductance was more closely correlated with xylem sap ABA concentration than with ABA flux or xylem sap pH and cation concentrations. In isohydric black walnut, increased concentrations of ABA in the xylem sap appeared to be primarily of root origin, causing stomatal closure in response to soil drying. In anisohydric sugar maple and white oak, however, there were reductions in midday leaf water potential associated with stomatal closure, making it uncertain whether drought-induced xylem sap ABA was of leaf or root origin. The role of root-originated xylem sap ABA in these species as a signal to the shoot of the water status of the roots is, therefore, less certain. PMID- 12651355 TI - Long-term photosynthetic acclimation to increased atmospheric CO(2) concentration in young birch (Betula pendula) trees. AB - To study the long-term response of photosynthesis to elevated atmospheric CO(2) concentration in silver birch (Betula pendula Roth.), 18 trees were grown in the field in open-top chambers supplied with 350 or 700 &mgr;mol mol(-1) CO(2) for four consecutive growing seasons. Maximum photosynthetic rates, stomatal conductance and CO(2) response curves were measured over the fourth growing season with a portable photosynthesis system. The photosynthesis model developed by Farquhar et al. (1980) was fitted to the CO(2) response curves. Chlorophyll, soluble proteins, total nonstructural carbohydrates, nitrogen and Rubisco activity were determined monthly. Elevated CO(2) concentration stimulated photosynthesis by 33% on average over the fourth growing season. However, comparison of maximum photosynthetic rates at the same CO(2) concentration (350 or 700 &mgr;mol mol(-1)) revealed that the photosynthetic capacity of trees grown in an elevated CO(2) concentration was reduced. Analysis of the response curves showed that acclimation to elevated CO(2) concentration involved decreases in carboxylation efficiency and RuBP regeneration capacity. No clear evidence for a redistribution of nitrogen within the leaf was observed. Down-regulation of photosynthesis increased as the growing season progressed and appeared to be related to the source-sink balance of the trees. Analysis of the main leaf components revealed that the reduction in photosynthetic capacity was accompanied by an accumulation of starch in leaves (100%), which was probably responsible for the reduction in Rubisco activity (27%) and to a lesser extent for reductions in other photosynthetic components: chlorophyll (10%), soluble protein (9%), and N concentrations (12%) expressed on an area basis. Despite a 21% reduction in stomatal conductance in response to the elevated CO(2) treatment, stomatal limitation was significantly less in the elevated, than in the ambient, CO(2) treatment. Thus, after four growing seasons exposed to an elevated CO(2) concentration in the field, the trees maintained increased photosynthetic rates, although their photosynthetic capacity was reduced compared with trees grown in ambient CO(2). PMID- 12651356 TI - Freezing behaviors in leaf buds of cold-hardy conifers visualized by NMR microscopy. AB - 1H-Nuclear magnetic resonance (NMR) microscopy was used to study freezing behavior in wintering leaf buds of Momi fir (Abies firma Sieb. et Zucc.) and Japanese red pine (Pinus densiflora Sieb. et Zucc.). The images acquired predominantly reflected the density of mobile (i.e., non-ice) protons mainly from unfrozen water. By comparing images taken at various subfreezing temperatures, we determined which tissues produced the high and low temperature exotherms detected by differential thermal analyses. Typical extra-organ freezing was successfully imaged in leaf buds of A. firma. The bud scales readily froze at -7 degrees C, but shoot primordia remained supercooled to -14 degrees C in December buds and to -21 degrees C in March buds. The size of supercooled shoot primordia was reduced with decreasing temperature, indicating a gradual decrease in water content of the shoot primordia. In contrast, the signal from shoot primordia of P. densiflora disappeared between -7 and -14 degrees C, corresponding to the high temperature exotherm at -8 degrees C, indicating extracellular freezing of the shoot primordia. The xylem and bark tissues readily froze at -7 degrees C in A. firma and between -7 and -14 degrees C in P. densiflora. We conclude that NMR microscopy can noninvasively provide more spatially specific information about freezing behavior in leaf buds than traditional methods such as differential thermal analysis. In particular, it allows the organized and harmonized freezing behaviors in complex organs to be visualized directly thereby revealing the diversity of mechanisms involved in freezing behaviors. PMID- 12651357 TI - Effects of current-year and previous-year PPFDs on shoot gross morphology and leaf properties in Fagus japonica. AB - We investigated how shoot gross morphology and leaf properties are determined in Fagus japonica Maxim., a deciduous species with flush-type shoot phenology, in which all leaves are produced in a single flush at the start of each season. We examined relationships between current-year shoot properties and local light environment in a 14-m tall beech tree growing in a deciduous forest. Leaf number (LN), total leaf area (TLA), and total leaf length (SL) of the current-year shoot increased with increasing photosynthetic photon flux density (PPFD). Leaf thickness, dry mass per leaf area and nitrogen content on a leaf area basis increased, whereas the chlorophyll/N ratio decreased with increasing PPFD. To separate the effects of current-year PPFD from those of previous year(s), we artificially shaded a part of the uppermost leaf tier. Reciprocal transfers of beech seedlings between controlled PPFD regimes were also made. Characteristics of shoot gross morphology such as LN, TLA and SL were largely determined by the PPFD of the previous year. The exception was the length of the longest "long shoots" with many leaves, in which elongation appeared to be influenced by both previous-year and current-year PPFD. In contrast, leaf properties were determined by current-year PPFD. The ecological implications of our findings are discussed. PMID- 12651358 TI - Stoichiometry of foliar carbon constituents varies along light gradients in temperate woody canopies: implications for foliage morphological plasticity. AB - Foliar morphology and chemical composition were examined along a light gradient in the canopies of five deciduous temperate woody species, ranked according to shade-tolerance as Populus tremula L. < Fraxinus excelsior L. < Tilia cordata Mill. = Corylus avellana L. < Fagus sylvatica L. Foliar carbon was divided between structural (cell-wall polysaccharides, lignin) and nonstructural (proteins, ethanol-soluble carbohydrates, starch) fractions. Foliar morphology of all species was strongly affected by irradiance. Both leaf dry mass per area (M(A)), a product of leaf density and thickness, and leaf dry to fresh mass ratio (D(w)), characterizing the apoplastic leaf fraction, increased with increasing relative irradiance (I(sum), calculated as the weighted mean of fractional penetration of diffuse and direct irradiance). Though the relationships were qualitatively identical among the taxa, more shade-tolerant species generally had lower values of M(A) than shade-intolerant species, and their morphological relationships with irradiance were curvilinear; however, there were no signs of saturation even at the highest irradiances in shade-intolerant species. In all species, lignin concentrations increased and cell-wall polysaccharide concentrations decreased with increasing irradiance. Consequently, biomass investment in structural leaf components appeared to be relatively constant along light gradients. The relationship between irradiance and structural compounds tended to be asymptotic in the more shade-tolerant species, whereas M(A) was linearly correlated with concentrations of structural leaf components, suggesting that similar factors were responsible for the curvature in the morphological and chemical relationships with irradiance. Because lignin increases tissue elastic modulus thereby rendering leaves more resistant to low leaf water potentials, we conclude that changes in stoichiometry of cell wall components were related to foliage acclimation to the gradients of water deficit that develop in the canopy and inherently accompany light gradients. We also conclude that increased lignification decreased leaf expansion growth, and that species differences in lignification were partly responsible for the observed interspecific variability in morphological plasticity. Analysis of structural leaf compounds provided no indication of how shade-intolerant species with low investments in lignin acclimated to gradients of water availability in the canopy. Because shade intolerant species generally had higher capacities for photosynthesis than shade tolerant species, we postulated that they should also have a greater ability for osmotic adjustment of leaf water potential with photosynthates. The concentrations of soluble carbohydrates increased with increasing irradiance in all species; however, the osmotic adjustment achieved in this way was similar in all species, except for shade-intolerant F. excelsior, which had a lower potential for osmotic adjustment with carbohydrates than the other taxa. Although we did not determine whether the gradients of stem water potential and leaf water deficits were similar in canopies of different species, we demonstrated that water relations play a central role in determining foliar structure and composition along light gradients in the canopy. PMID- 12651359 TI - Interactive effects of nitrogen and water availabilities on gas exchange and whole-plant carbon allocation in poplar. AB - Cuttings of balsam spire hybrid poplar (Populus trichocarpa var. Hastata Henry x Populus balsamifera var. Michauxii (Dode) Farwell) were grown in sand culture and irrigated every 2 (W) or 10 (w) days with a solution containing either 3.0 (N) or 0.5 (n) mol nitrogen m(-3) for 90 days. Trees in the WN (control) and wn treatments had stable leaf nitrogen concentrations averaging 19.4 and 8.4 mg g( 1), respectively, over the course of the experiment. Trees in the Wn and wN treatments had a similar leaf nitrogen concentration, which increased from 12.0 to 15.8 mg g(-1) during the experiment. By the final harvest, mean stomatal conductances of trees in the wN and wn treatments were less than those of trees in the Wn and WN treatments (1.8 versus 4.6 mm s(-1)). Compared to the WN treatment, biomass at the final harvest was reduced by 61, 72 and 75% in the Wn, wN and wn treatments, respectively. At the final harvest, WN trees had a mean total leaf area of 4750 +/- 380 cm(2) tree(-1) and carried 164 +/- 8 leaves tree( 1) with a specific leaf area of 181 +/- 16 cm(2) g(-1), whereas Wn trees had a smaller mean total leaf area (1310 +/- 30 cm(2) tree(-1)), because of the production of fewer leaves (41 +/- 6) with a smaller specific leaf area (154 +/- 2 cm(2) g(-1)). A greater proportion of biomass was allocated to roots in Wn trees than in WN trees, but component nitrogen concentrations adjusted such that there was no Wn treatment effect on nitrogen allocation. Compared with WN trees, rates of photosynthesis and respiration per unit weight of tissue of Wn trees decreased by 28 and 31%, respectively, but the rate of photosynthesis per unit leaf nitrogen remained unaltered. The wN and Wn trees had similar leaf nitrogen concentrations; however, compared with the Wn treatment, the wN treatment decreased mean total leaf area (750 +/- 50 cm(2) tree(-1)), number of leaves per tree (29 +/- 2) and specific leaf area (140 +/- 6 cm(2) g(-1)), but increased the allocation of biomass and nitrogen to roots. Net photosynthetic rate per unit leaf nitrogen was 45% lower in the wN treatment than in the other treatments. Rates of net photosynthesis and respiration per unit weight of tissue were 48 and 33% less, respectively, in wN trees than in Wn trees. PMID- 12651360 TI - A carbon balance model of peach tree growth and development for studying the pruning response. AB - We modeled tree responses to pruning on the basis of growth rules established on unpruned trees and a simple principle governing root-shoot interactions. The model, which integrates architectural and ecophysiological approaches, distinguishes four types of anatomical organs in a tree: rootstock, main axis, secondary axes and new roots. Tree structure is described by the position of secondary axes on the main axis. The main processes considered are plastochronal activity, branching, assimilate production, respiration and assimilate partitioning. Growth and development rules were based on measurements of two unpruned trees. The model was used to simulate growth of peach trees (Prunus persica (L.) Batsch) in their first growing season. Assuming that the equilibrium between roots and shoots tends to be restored after pruning, the response to removal of the main axis above the twentieth internode in mid-July was simulated and compared to the response measured in three pruned trees. The model fit the unpruned tree data reasonably well and predicted the main traits of tree behavior after pruning. Dry matter growth of the secondary axes of pruned trees was increased so that shoot seasonal carbon balance was hardly modified by pruning. Rhythmicity of growth was enhanced by pruning, and might result from variations induced in the root:shoot ratio. Variation in pruning severity had greater effects than variation in pruning date. A sensitivity analysis indicated that: (1) root-shoot partitioning was a critical process of the model; (2) tree growth was mainly dependent on assimilate availability; and (3) tree shape was highly dependent on the branching process. PMID- 12651361 TI - Water deficits are more important in delaying growth than in changing patterns of carbon allocation in Eucalyptus globulus. AB - Potted cuttings of three Eucalyptus globulus Labill. clones (AR3, CN44, MP11) were either well watered or subjected to one of two soil water deficit regimes for six months in a greenhouse. Reductions in lateral branching, leaf production and leaf expansion were the leading contributors to the large differences observed in biomass production between well-watered and water-stressed plants. Although no significant differences among clones were observed in dry matter accumulation or in the magnitude of the response to soil water deficits, sensitivity of lateral branching, leaf initiation and whole-plant foliage to water stress was significantly lower in CN44 than in AR3 and MP11. When the confounding effect of differences in plant size resulting from the different watering regimes was removed, allometric analysis indicated that the genotypes differed in biomass allocation patterns. In addition to a drought-induced reduction in leaf number, water deficits also resulted in smaller leaves because leaf expansion was inhibited during dehydration events. Resumption of leaf expansion following stress relief occurred in all of the clones, but was particularly evident in severely stressed plants of Clone AR3, possibly as a result of the osmotic adjustment observed in this genotype. PMID- 12651362 TI - Regulation of ponderosa pine foliar physiology and insect resistance mechanisms by basal area treatments. AB - We compared foliar physiology and several measures of tree resistance to insect attack among ponderosa pine (Pinus ponderosa var. scopulorum Engelm.) trees growing in thinned stands. Measurements were made in a second-growth ponderosa pine forest in northern Arizona where the basal area treatments (6.9, 18.4, 27.6, 78.2 m(2) ha(-1)) have been experimentally maintained by frequent thinnings for 32 years before our measurements began in 1994. Most of the physiological characteristics measured were affected by the basal area treatments. As stand basal area increased from 6.9 to 78.2 m(2) ha(-1), predawn water potential, midday water potential, net photosynthetic rate, resin production, phloem thickness, and foliar toughness decreased. Foliar nitrogen concentration was greatest in trees in the intermediate basal area treatments. Our results show that the physiological condition of second-growth ponderosa pine can be manipulated by silvicultural control of stand basal area, and support the hypothesis that high stand basal area increases tree stress and decreases tree resistance to insect attack. PMID- 12651363 TI - Ethylene evolution, radial growth and carbohydrate concentrations in Abies balsamea shoots ringed with Ethrel. AB - The terminal (1-year-old) shoot of quiescent, 2-year-old Abies balsamea (L.) Mill. seedlings was either left untreated or ringed with 0, 1 or 10 mg Ethrel g( 1) lanolin. After 5 weeks of culture under environmental conditions favorable for growth, the shoots were harvested to measure ethylene evolution and carbohydrate concentrations by gas chromatography, and tracheid number and bark radial width by microscopy. In untreated shoots, the basal rate of ethylene evolution followed the order: cambial region > cortex + periderm = xylem + pith = needles. Wound induced ethylene production was not detected until at least 4 h after excision, but was evident in all fractions 24 h after excision; the increase in wound induced ethylene evolution followed the order: cambial region > cortex + periderm > xylem + pith > needles. Compared with untreated controls, the application of plain lanolin, which involved the removal of needles and periderm, increased bark radial width and wound-induced ethylene production by the cambial region and the cortex + periderm, but decreased cambial region concentrations of fructose, glucose and starch at the application point. At the application point, Ethrel concomitantly increased ethylene evolution from the cambial region and the cortex + periderm, tracheid number, bark radial width, and the cambial region concentrations of fructose, glucose, sucrose and starch. No effects of Ethrel treatment were detected above or below the application point, with the exception that the 10 mg g(-1) Ethrel treatment stimulated ethylene evolution and decreased starch concentration of the cambial region. The results indicate that: (1) the cambial region is the major source of endogenous ethylene in the 1-year-old shoot; (2) the magnitude of the difference in ethylene evolution between particular shoot fractions is different before and after the start of wound induced ethylene production; (3) the Ethrel-induced increase in tracheid number and bark radial width at the application point is positively related to ethylene evolution from the cambial region and the cortex + periderm, respectively; and (4) ethylene derived from Ethrel applied laterally to a woody stem can mobilize carbohydrates to the application point. PMID- 12651364 TI - The response of Pinus sylvestris to drought: stomatal control of transpiration and hydraulic conductance. AB - We investigated the impact of drought on the physiology of 41-year-old Scots pine (Pinus sylvestris L.) in central Scotland. Measurements were made of the seasonal course of transpiration, canopy stomatal conductance, needle water potential, xylem water content, soil-to-needle hydraulic resistance, and growth. Comparison was made between drought-treated plots and those receiving average precipitation. In response to drought, transpiration rate declined once volumetric water content (VWC) over the top 20 cm of soil reached a threshold value of 12%. Thereafter, transpiration was a near linear function of soil water content. As the soil water deficit developed, the hydraulic resistance between soil and needles increased by a factor of three as predawn needle water potential declined from -0.54 to -0.71 MPa. A small but significant increase in xylem embolism was detected in 1-year old shoots. Stomatal control of transpiration prevented needle water potential from declining below -1.5 MPa. Basal area, and shoot and needle growth were significantly reduced in the drought treatment. In the year following the drought, canopy stomatal conductance and soil-to-needle hydraulic resistance recovered. Current-year needle extension recovered, but a significant reduction in basal area increment was evident one year after the drought. The results suggest that, in response to soil water deficit, mature Scots pine closes its stomata sufficiently to prevent the development of substantial xylem embolism. Reduced growth in the year after a severe soil water deficit is most likely to be the result of reduced assimilation in the year of the drought, rather than to any residual embolism carried over from one year to the next. PMID- 12651365 TI - Transport, storage and mobilization of nitrogen by trees and shrubs in the wet/dry tropics of northern Australia. AB - Xylem sap from woody species in the wet/dry tropics of northern Australia was analyzed for N compounds. At the peak of the dry season, arginine was the main N compound in sap of most species of woodlands and deciduous monsoon forest. In the wet season, a marked change occurred with amides becoming the main sap N constituents of most species. Species from an evergreen monsoon forest, with a permanent water source, transported amides in the dry season. In the dry season, nitrate accounted for 7 and 12% of total xylem sap N in species of deciduous and evergreen monsoon forests, respectively. In the wet season, the proportion of N present as nitrate increased to 22% in deciduous monsoon forest species. These results suggest that N is taken up and assimilated mainly in the wet season and that this newly assimilated N is mostly transported as amide-N (woodland species, monsoon forest species) and nitrate (monsoon forest species). Arginine is the form in which stored N is remobilized and transported by woodland and deciduous monsoon forest species in the dry season. Several proteins, which may represent bark storage proteins, were detected in inner bark tissue from a range of trees in the dry season, indicating that, although N uptake appears to be limited in the dry season, the many tree and shrub species that produce flowers, fruit or leaves in the dry season use stored N to support growth. Nitrogen characteristics of the studied species are discussed in relation to the tropical environment. PMID- 12651366 TI - Patterns of photosynthesis and starch allocation in seedlings of four bottomland hardwood tree species subjected to flooding. AB - Effects of short-term (32 days) flooding on photosynthesis, stomatal conductance, relative growth rate and tissue starch concentrations of flood-intolerant Quercus alba L. (white oak), bottomland Quercus nigra L. (water oak), bottomland Fraxinus pennsylvanica Marshall. (green ash) and flood-tolerant Nyssa aquatica L. (water tupelo) seedlings were studied under controlled conditions. Net photosynthetic rates of flooded N. aquatica seedlings were reduced by 25% throughout the 32-day flooding period. Net photosynthetic rates of flooded Q. alba seedlings fell rapidly to 25% of those of the control seedlings by Day 4 of the flooding treatment and to 5% by Day 16. In F. pennsylvanica and Q. nigra, net photosynthetic rates were reduced to 50% of control values by Day 8 but remained at approximately 30 and 23%, respectively, of control values by Day 32. Leaves of flooded Q. alba seedlings accumulated approximately twice as much starch as leaves of non-flooded control plants, whereas root starch concentrations decreased to 67% of those of control plants by the end of the 32-day flooding treatment. In contrast, flooding caused only a small increase in leaf starch concentrations of N. aquatica plants, but it increased root starch concentrations to 119% of those of the control plants by the end of the experiment. The co occurring bottomland species, Fraxinus pennsylvanica and Q. nigra, differed from each other in their patterns of stomatal conductance and root starch concentrations. We conclude that the maintenance of low leaf starch concentrations, and high pre-flood root tissue starch concentrations are important characteristics allowing flood-tolerant species to survive in flooded soils. PMID- 12651367 TI - Leaf phenology, photosynthesis, and the persistence of saplings and shrubs in a mature northern hardwood forest. AB - We quantified leaf phenologies of saplings and overstory trees of sugar maple (Acer saccharum Marsh.) and American beech (Fagus grandifolia Ehrh.), and the shrub hobblebush viburnum (Viburnum alnifolium Marsh.) in a 72-year-old northern hardwood forest. Seasonal changes in irradiance in the shrub layer, and in the leaf CO(2) exchange of viburnum, and sugar maple and beech saplings were also measured. Leaf expansion occurred earlier in the spring and green leaves were retained later in the autumn in saplings and shrubs than in overstory trees. During the spring light phase (before overstory closure), large CO(2) gains by all three shrub-layer species occurred as a result of a combination of relatively large leaf area, high photosynthetic capacity, and high irradiance. Throughout the summer shade phase, photosynthetic capacity at a given irradiance remained relatively constant, but CO(2) gain was typically limited by low irradiances. Even though irradiance in the shrub layer increased during the autumn light phase as the overstory opened, CO(2) gains were modest compared to springtime values because of declining leaf area and photosynthetic capacity in all three species. The CO(2) gains during the spring light phase, and to a lesser extent during the autumn light phase, may be important to the carbon balance and long-term persistence of saplings and shrubs in the usually light-limited shrub layer of a northern hardwood forest. Therefore, for some late-successional species, leaf phenology may be an important characteristic that permits their long-term persistence in the shrub layer of mature northern hardwood forests. PMID- 12651368 TI - Osmotic potential of several hardwood species as affected by manipulation of throughfall precipitation in an upland oak forest during a dry year. AB - Components of dehydration tolerance, including osmotic potential at full turgor (Psi(pio)) and osmotic adjustment (lowering of Psi(pio)), of several deciduous species were investigated in a mature, upland oak forest in eastern Tennessee. Beginning July 1993, the trees were subjected to one of three throughfall precipitation treatments: ambient, ambient minus 33% (dry treatment), and ambient plus 33% (wet treatment). During the dry 1995 growing season, leaf water potentials of all species declined to between -2.5 and -3.1 MPa in the dry treatment. There was considerable variation in Psi(pio) among species (-1.0 to 2.0 MPa). Based on Psi(pio) values, American beech (Fagus grandifolia Ehrh.), dogwood (Cornus florida L.), and sugar maple (Acer saccharum Marsh.) were least dehydration tolerant, red maple (A. rubrum L.) was intermediate in tolerance, and white oak (Quercus alba L.) and chestnut oak (Quercus prinus L.) were most tolerant. During severe drought, overstory chestnut oak and understory dogwood, red maple and chestnut oak displayed osmotic adjustment (-0.12 to -0.20 MPa) in the dry treatment relative to the wet treatment. (No osmotic adjustment was evident in understory red maple and chestnut oak during the previous wet year.) Osmotic potential at full turgor was generally correlated with leaf water potential, with both declining over the growing season, especially in species that displayed osmotic adjustment. However, osmotic adjustment was not restricted to species considered dehydration tolerant; for example, dogwood typically maintained high Psi(pio) and displayed osmotic adjustment to drought, but had the highest mortality rates of the species studied. Understory saplings tended to have higher Psi(pio) than overstory trees when water availability was high, but Psi(pio) of understory trees declined to values observed for overstory trees during severe drought. We conclude that Psi(pio) varies among deciduous hardwood species and is dependent on canopy position and soil water potential in the rooting zone. PMID- 12651369 TI - Water relations of several hardwood species in response to throughfall manipulation in an upland oak forest during a wet year. AB - We investigated the effects of altered precipitation on leaf osmotic potential at full turgor (Psi(pio)) of several species in an upland oak forest during the 1994 growing season as part of a Throughfall Displacement Experiment at the Walker Branch Watershed near Oak Ridge, Tennessee. The main species sampled included overstory chestnut oak (Quercus prinus L.), white oak (Q. alba L.), red maple (Acer rubrum L.); intermediates sugar maple (A. saccharum L.) and blackgum (Nyssa sylvatica Marsh.); and understory dogwood (Cornus florida L.) and red maple. The precipitation treatments were: ambient precipitation; ambient minus 33% of throughfall (dry); and ambient plus 33% of throughfall (wet). Except in late September, midday leaf water potentials (Psi(l)) were generally high in all species in all treatments, ranging from -0.31 to -1.34 MPa for C. florida, -0.58 to -1.51 MPa for A. rubrum, and -0.78 to -1.86 MPa for Q. prinus. Both treatment and species differences in Psi(pio) were evident, with oak species generally exhibiting lower Psi(pio) than A. saccharum, A. rubrum, C. florida, and N. sylvatica. The Psi(pio) of C. florida saplings declined in the dry treatment, and Q. prinus, Q. alba, and A. saccharum all exhibited a declining trend of Psi(pio) in the dry treatment, although Psi(pio) of Q. prinus leaves increased in late August, corresponding to a recovery in soil water potential. Cornus florida exhibited osmotic adjustment with the largest adjustment coinciding with the period of lowest soil water potential in June. The only other species to exhibit osmotic adjustment was Q. prinus, which also maintained a lower baseline Psi(pio) than the other species. We conclude that a 33% reduction of throughfall is sufficient both to alter the water relations of some species in the upland oak forest and to enable the identification of those species capable of osmotic adjustment to a short-term drought during a wet year. PMID- 12651370 TI - Canopy conductance of Pinus taeda, Liquidambar styraciflua and Quercus phellos under varying atmospheric and soil water conditions. AB - Sap flow, and atmospheric and soil water data were collected in closed-top chambers under conditions of high soil water potential for saplings of Liquidambar styraciflua L., Quercus phellos L. and Pinus taeda L., three co occurring species in the southeastern USA. Responses of canopy stomatal conductance (g(t)) to water stress induced by high atmospheric water vapor demand or transpiration rate were evaluated at two temporal scales. On a diurnal scale, the ratio of canopy stomatal conductance to maximum conductance (g(t)/g(t,max)) was related to vapor pressure deficit (D), and transpiration rate per unit leaf area (E(l)). High D or E(l) caused large reductions in g(t)/g(t,max) in L. styraciflua and P. taeda. The response of g(t)/g(t,max) to E(l) was light dependent in L. styraciflua, with higher g(t)/g(t,max) on sunny days than on cloudy days. In both L. styraciflua and Q. phellos, g(t)/g(t,max) decreased linearly with increasing D (indicative of a feed-forward mechanism of stomatal control), whereas g(t)/g(t,max) of P. taeda declined linearly with increasing E(l) (indicative of a feedback mechanism of stomatal control). Longer-term responses to depletion of soil water were observed as reductions in mean midday g(t)/g(t,max), but the reductions did not differ significantly between species. Thus, species that employ contrasting methods of stomatal control may show similar responses to soil water depletion in the long term. PMID- 12651371 TI - Photosynthetic gas exchange and accumulation of phytotoxins in mangrove seedlings in response to soil physico-chemical characteristics associated with waterlogging. AB - We evaluated photosynthetic gas exchange and the accumulation of iron, manganese and sulfur in seedlings of five mangrove species (Aegiceras corniculatum (L.) Blanco, Avicennia marina (Forsk.) Vierh., Bruguiera gymnorrhiza (L.) Lamk., Hibiscus tiliaceus L. and Rhizophora stylosa Griff.) growing under anoxic soil conditions at low irradiance. Seedlings of the viviparous mangroves showed no significant responses to root anoxia. The presence of ferrous sulfate or manganous sulfate had a smaller effect on CO(2) assimilation, transpiration rate and stomatal behavior than the presence of sodium sulfide. Sodium sulfide inhibited photosynthetic gas exchange and caused complete stomatal closure in all species. Stomatal closure was probably the result of the damaging effect of sulfide ions on root cell membranes. Some leaf epinasty and wilting were also observed in response to the Na(2)S treatment in all species. A combination of FeSO(4), MnSO(4) and Na(2)S had a smaller effect on photosynthesis than Na(2)S alone, especially for Avicennia marina and Rhizophora stylosa seedlings, which maintained appreciable rates of CO(2) assimilation (2.49 and 3.84 &mgr;mol m(-2) s(-1), respectively) in the presence of all three phytotoxins. Roots of phytotoxin-treated seedlings of all species accumulated significant amounts of the corresponding toxin compared with roots of the control plants. The FeSO(4) or MnSO(4) treatments had no effect on foliar concentrations of iron or manganese, whereas the Na(2)S treatment resulted in an accumulation of S in the leaves of all species. Interactions between Fe(2+) and Mn(2+) and sulfide in the culture medium appeared to reduce their uptake by the seedlings. We conclude that high concentrations of sulfides in mangrove sediments may adversely affect both growth and survival of mangrove seedlings at low irradiances. PMID- 12651372 TI - Effect of season and soil treatments on carbohydrate concentrations in Norway spruce (Picea abies) mycorrhizae. AB - We studied effects of season and soil treatments (watering, acidification, liming and combinations of these treatments) on soluble carbohydrates of mycorrhizal roots of Norway spruce (Picea abies (L.) Karst.). Arabinose, arabitol, fructose, glucose, inositol, lactose, mannitol, pinite, quinate, raffinose, shikimate, stachyose and trehalose were identified by HPLC. Concentrations of inositol, lactose and pinite were constant throughout the year, whereas concentrations of raffinose, stachyose and trehalose were higher in winter than in summer, and concentrations of glucose, fructose and mannitol increased from February to September. Soil acidification and liming had no effect on the annual mean concentrations of fructose, glucose, lactose, pinite, raffinose and stachyose. Liming increased quinate concentrations and decreased arabitol concentrations. Annual mean concentrations of arabinose and mannitol decreased in response to soil acidification. Annual mean concentrations of inositol increased in response to irrigation. None of the soil treatments affected the mean annual concentration of trehalose. PMID- 12651373 TI - Hydraulic conductivity in roots of ponderosa pine infected with black-stain (Leptographium wageneri) or annosus (Heterobasidion annosum) root disease. AB - Roots from healthy and diseased mature ponderosa pine, Pinus ponderosa Laws., trees were excavated from a site near Burns, Oregon. The diseased trees were infected with black-stain root disease, Leptographium wageneri Kendrick, or annosus root disease, Heterobasidion annosum (Fr.) Bref., or both. Axial hydraulic conductivity of the roots was measured under a positive head pressure of 5 kPa, and the conducting area was stained with safranin dye to determine specific conductivity (k(s)). In diseased roots, only 8-12% of the cross sectional xylem area conducted water. Resin-soaked xylem completely restricted water transport and accounted for 13-16% of the loss in conducting area. In roots with black-stain root disease, 17% of the loss in conducting area was associated with unstained xylem, possibly resulting from occlusions or embolisms. Based on the entire cross-sectional area of infected roots, the k(s) of roots infected with black-stain root disease was 4.6% of that for healthy roots, whereas the k(s) of roots infected with annosus root disease was 2.6% of that for healthy roots. Although these low values were partly the result of the presence of a large number of diseased roots (72%) with no conducting xylem, the k(s) of functional xylem of diseased roots was only 33% of that for healthy roots. The low k(s) values of functional xylem in diseased roots may be caused by fungus induced occlusions preceding cavitation and embolism of tracheids. The k(s) of disease-free roots from diseased trees was only 70% of that for healthy roots from healthy trees. The disease-free roots had the same mean tracheid diameter and tissue density as the healthy roots, suggesting that the lower k(s) in disease-free roots of diseased trees may also have been caused by partial xylary occlusions. PMID- 12651374 TI - Photosynthetic characteristics of Flindersia brayleyana and Castanospermum australe from tropical lowland and upland sites. AB - Photosynthetic responses to temperature, light and carbon dioxide partial pressure were studied in two-year-old Flindersia brayleyana F. Muell. and Castanospermum australe Cunn. & C. Fraser ex Hook. growing on coastal lowland and upland rainforest sites in tropical Queensland, Australia. Climatic conditions ranged from moist and cool (17-19 degrees C) to dry and warm (22-24 degrees C). The optimum temperature for photosynthesis was 23.7-25.6 degrees C for C. australe and 21.2-24.6 degrees C for F. brayleyana. Mean maximum rate of electron transport for each species did not differ between sites but was higher (60-62 &mgr;mol m(-2) s(-1)) in F. brayleyana than in C. australe (42-44 &mgr;mol m(-2) s(-1)). Ribulose-bisphosphate carboxylation rate did not differ significantly between sites or species. Maximum rates of photosynthesis at 1000 &mgr;Pa Pa(-1) CO(2) did not differ significantly between sites for each species, but did differ significantly between species. At 350 &mgr;Pa Pa(-1) CO(2), photosynthetic light use efficiencies of F. brayleyana and C. australe were 0.05 and 0.015, respectively, at the upland site, and the corresponding values at the lowland site were 0.025 and 0.05. In C. australe, these differences were reflected in significantly greater maximum rates of photosynthesis at 350 &mgr;Pa Pa(-1) CO(2) at the lowland site than at the upland site (5.2 versus 3.3 &mgr;mol m(-2) s( 1)). PMID- 12651375 TI - Dynamics of change in stomatal response and water status of Picea abies during a persistent drought period: a contribution to the traditional view of plant water relations. AB - Four experiments on the simulation of a persistent drought period were carried out with cloned Picea abies (L.) Karst. trees: two in the field under varying weather conditions and two in a climate chamber under variously manipulated humidity conditions. Patterns of diurnal dynamics in gas exchange rates and water potential were monitored and analyzed. The first phase of the drought was characterized by relatively high daily maxima for photosynthesis and transpiration. With decreasing humidity during the day, the values dropped steeply, and the declines were larger and occurred earlier on each passing day of the drought period. When soil water potential was lower than -2000 hPa, maximum stomatal aperture was greatly reduced despite a humid atmosphere. Under these conditions, rates of photosynthesis and transpiration decreased less steeply from the daily maxima and differences between maxima and minima were small. In the field, the daily sums of transpiration and photosynthesis were more dependent on atmospheric conditions than on soil water potential. In the growth chamber experiments, the daily sums of transpiration and photosynthesis decreased continuously as the soil dried, at first steeply until a soil water potential of 2000 hPa was reached, then slowly. Predawn water potential values fluctuated under field conditions, but tended to decrease with time, whereas needle osmotic potential increased slightly. Because relative humidities did not reach 100% in the growth chamber, predawn water potentials of plants in the growth chamber were never higher than -1.0 MPa although the soil was saturated. In the experiment with a high average air humidity during the daily stress period, relatively high predawn water potentials were maintained until lower soil water potentials of 8000 hPa were reached. Results were used to assess the importance of evaporative demand versus soil drying on stomatal responses within the context of current concepts of plant water relations. The observed trends in diurnal dynamics can be explained solely by the interdependency of leaf conductance and water potential. Stomata react directly to the ratio of water supply to demand. The central role of peristomatal transpiration in this system is emphasized. PMID- 12651376 TI - Variability of the photosynthetic characteristics of mature needles within the crown of a 25-year-old Pinus pinaster. AB - Photosynthetic characteristics of 1- and 2-year-old needles were determined in excised shoots of maritime pine (Pinus pinaster Ait.) with an open gas exchange system. We used the nonlinear least mean squares method to derive values for quantum yield of electron transport (alpha), maximum carboxylation velocity (V(cmax)), and maximum electron transport rate (J(max)), from photosynthetic response curves to light and CO(2). Crown height had no significant effect on any of the parameters; however, V(cmax) and J(max), as well as alpha were 43, 26 and 35% higher, respectively, in 1-year-old needles than in 2-year-old needles. The main effect of irradiance on needles was a small decline in leaf concentrations of nitrogen and phosphorus from the top to the bottom of the canopy. Only J(max) demonstrated a linear relationship with both nitrogen content (R(2) = 0.42) and irradiance at the shoot level. Because needle age accounted for most of the variability in photosynthesis, we incorporated needle age into the photosynthesis model of Farquhar et al. (1980). The modified model underestimated the daily assimilation rate of 1-year-old needles in the field, especially when assimilation rates were high. PMID- 12651377 TI - Effects of elevated CO(2) concentration and nutrition on net photosynthesis, stomatal conductance and needle respiration of field-grown Norway spruce trees. AB - To study the effects of elevated CO(2) on gas exchange, nonstructural carbohydrate and nutrient concentrations in current-year foliage of 30-year-old Norway spruce (Picea abies (L.) Karst.) trees, branches were enclosed in ventilated, transparent plastic bags and flushed with ambient air (mean 370 &mgr;mol CO(2) mol(-1); control) or ambient air + 340 &mgr;mol CO(2) mol(-1) (elevated CO(2)) during two growing seasons. One branch bag was installed on each of 24 selected trees from control and fertilized plots. To reduce the effect of variation among trees, results from each treated branch were compared with those from a control branch on the same whorl of the same tree. Elevated CO(2) increased rates of light-saturated photosynthesis on average by 55% when measured at the treatment CO(2) concentration. The increase was larger in shoots with high needle nitrogen concentrations than in shoots with low needle nitrogen concentrations. However, shoots grown in elevated CO(2) showed a decrease in photosynthetic capacity compared with shoots grown in ambient CO(2). When measured at the internal CO(2) concentration of 200 &mgr;mol CO(2) mol(-1), photosynthetic rates of branches in the elevated CO(2) treatments were reduced by 8 to 32%. The elevated CO(2) treatment caused a 9 to 20% reduction in carboxylation efficiency and an 18% increase in respiration rates. In response to elevated CO(2), starch, fructose and glucose concentrations in the needles increased on average 33%, whereas concentrations of potassium, nitrogen, phosphorus, magnesium and boron decreased. Needle nitrogen concentrations explained 50-60% of the variation in photosynthesis and CO(2) acclimation was greater at low nitrogen concentrations than at high nitrogen concentrations. We conclude that the enhanced photosynthetic rates found in shoots exposed to elevated CO(2) increased carbohydrate concentrations, which may have a negative feedback on the photosynthetic apparatus and stimulate cyanide-resistant respiration. We also infer that the decrease in nutrient concentrations of needles exposed to elevated CO(2) was the result of retranslocation of nutrients to other parts of the branch or tree. PMID- 12651378 TI - Seasonal fluctuations of starch in root and stem tissues of coppiced Salix viminalis plants grown under two nitrogen regimes. AB - Seasonal changes in starch were studied at the tissue and cellular levels in roots and stems of Salix viminalis L. cuttings. Cuttings were planted in pots containing sand and grown in a controlled environment chamber in which seasons were artificially induced by changes in temperature and photoperiod. Nitrogen was supplied at optimum and low rates, and during dormancy, one-half of the plants were decapitated. Starch concentrations in root and stem tissues were determined regularly during shoot extension growth, dormancy and resprouting after dormancy. We used light microscopy (LM) combined with image analysis (IA) to determine the cellular localization and amount of starch in different cell types of stem and root tissues. Chemical analysis confirmed that starch concentrations were lower in plants receiving a high-N supply rate than in plants receiving a low-N supply rate. In all plants, the highest concentration of starch was in the roots. Light microscopy and IA showed that starch accumulated mainly in the phloem and cortical cells of both root and stem tissues. Starch grains were also regularly found in ray parenchyma cells. The amount of starch as well as the size of the grains showed strong seasonal fluctuations. In both roots and stems, starch concentrations were highest during predormancy and lowest during periods of shoot extension growth. At the time of resprouting, root cells of decapitated plants were more depleted of starch than root cells of intact plants, supporting the hypothesis that starch reserves in roots are important during the early phase of resprouting in coppice systems. PMID- 12651379 TI - Morphological and physiological markers of juvenility and maturity in shoot cultures of oak (Quercus robur and Q. petraea). AB - Evaluation of potential techniques for rejuvenating oak shoots requires robust quantitative markers for juvenile and mature plants. To identify suitable in vitro markers, shoot cultures of Quercus species of juvenile, adolescent and mature origin were screened for a range of morphological and physiological markers of juvenility and maturity. Criteria examined included angle of the shoot to the horizontal, stem length, stem diameter (tip, mid, base), leaf number, scale leaf number and shoot number. Image analysis was also carried out to determine leaf area, size, perimeter and breadth and length of leaves. Mature Q. robur L. clones had a larger mid-stem and tip diameter than juvenile clones, whereas mature Q. petraea ex Liebl. clones were characterized by plagiotropic growth and larger mid-stem and tip diameters compared with juvenile clones. Based on discriminant analysis of the data, we propose the following formulae for discrimination of juvenile and mature shoots, where a negative value for the discriminant score (D) indicates juvenility. For Q. petraea: D = -1.308 - 0.0351 SA + 2.206 TD + 1.435 MSD, where SA is stem angle ( degrees ), TD is tip diameter (mm) and MSD is mid-stem diameter (mm). For Q. robur: D = -3.546 + 2.418 TD + 2.202 MSD. Quercus robur clones derived from stump sprouts and designated as juvenile had a negative D value suggesting a juvenile status for these clones. Clones sourced from a hedged, grafted Q. robur tree of mature origin had a positive D value indicating a mature status. Clones initiated from a 20-25-year old Q. petraea tree displayed morphology in vitro consistent with a mature status and had a positive D value; however, these clones displayed other traits such as vigor suggesting that vestiges of juvenility remain. Multiplication rate and leaf size and shape were variable among clones and did not provide suitable markers for juvenility or maturity for these Quercus species. PMID- 12651380 TI - Differences in root longevity of some tree species. AB - Although the importance of root production and mortality to nutrient fluxes in ecosystems is widely recognized, the difficulties associated with root measurements have limited the availability of reliable data. We have used minirhizotrons and image analysis to measure root longevity of Prunus avium L., Picea sitchensis (Bong.) Carriere, Acer pseudoplatanus L. and Populus x canadensis cv. Beaupre directly in cohorts of roots. Major differences in the longevity of roots among species were identified. For example, 40% of Prunus avium roots but only 6% of Picea sitchensis roots survived for more than 14 days. Survival analysis of cohorts of roots of Prunus avium and Populus x canadensis revealed differences in the distribution of longevity among cohorts. Genetic, biotic and abiotic factors that may influence longevity are discussed. PMID- 12651381 TI - Variations in the secondary metabolite camptothecin in relation to tissue age and season in Camptotheca acuminata. AB - We investigated variation in concentration of the secondary metabolite, camptothecin (CPT), in relation to leaf, branch and tree age, season, and leaf drying method in Camptotheca acuminata Decaisne saplings. Younger leaves contained higher CPT concentrations than older leaves. Within a branch, there was a linear decline in CPT concentration from leaves at the apex of the branch down to Leaf 7. Comparing leaves of similar age, those from younger trees had higher CPT concentrations than those from older trees. Over the course of the growing season, there was a steady decline of 11% per month in leaf CPT concentration. Branches showed a similar seasonal decline in CPT concentration to leaves; however, the rate of decline was threefold greater in leaves than in branches. Freeze-dried tissues had a 27% higher CPT concentration than oven- or air-dried tissues, suggesting that oven- and air-drying caused degradation of CPT. The decline in CPT concentration with tissue aging may reflect a genetically determined mechanism whereby, in young trees, chemicals serve as a first line of defense against attacks by herbivores and pathogenic microorganisms until other mechanisms are developed and deployed. We hypothesize that chemical defense mechanisms are programmed for early ontogenic stages, whereas they are induced by biotic and abiotic factors during later ontogenic stages. PMID- 12651382 TI - Photosynthetic capacity of red spruce during winter. AB - We measured the photosynthetic capacity (P(max)) of plantation-grown red spruce (Picea rubens Sarg.) during two winter seasons (1993-94 and 1994-95) and monitored field photosynthesis of these trees during one winter (1993-94). We also measured P(max) for mature montane trees from January through May 1995. Changes in P(max) and field photosynthesis closely paralleled seasonal changes in outdoor air temperature. However, during thaw periods, field photosynthesis was closely correlated with multiple-day temperature regimes, whereas P(max) was closely correlated with single-day fluctuations in temperature. There was a strong association between short-term changes in ambient temperature and P(max) during the extended thaw of January 1995. Significant increases in P(max) occurred within two days of the start of this thaw. Repeated measurements of cut shoots kept indoors indicated that temperature-induced increases in P(max) can occur within 3 h. Although significant correlations between P(max) and stomatal conductance (g(s)) or intracellular CO(2) concentration (C(i)) raised the possibility that increases in P(max) resulted from increases in stomatal aperture, fluctuations in g(s) or C(i) explained little of the overall variation in P(max). Following both natural and simulated thaws, P(max) increased considerably but plateaued at only 37% of the mean photosynthetic rate reported for red spruce during the growing season. Thus, even though shoots were provided with near-optimal environmental conditions, and despite thaw-induced changes in physiology, significant limitations to winter photosynthesis remained. PMID- 12651383 TI - Comparative seed ecophysiology of wild and cultivated Carica papaya trees from a tropical rain forest region in Mexico. AB - To ascertain the effects of centuries of cultivation practices on seed behavior and dormancy mechanisms, we compared seed size and germination characteristics of wild and cultivated (domesticated) populations of Carica papaya L. Germination experiments were carried out under various conditions of temperature, light, seed soaking and gibberellic acid treatments. Wild papaya seeds showed responses to treatment that are characteristic of seeds of many rain forest pioneer trees. Seeds were small and light sensitive, whereas cultivated papaya seeds were 33% larger and their light responses as well as other physiological traits indicated that cultivation had resulted in a lessening in the importance of specific environmental conditions for dormancy breaking and germination. PMID- 12651384 TI - Parameterization and testing of a coupled photosynthesis-stomatal conductance model for boreal trees. AB - A coupled photosynthesis-stomatal conductance model was parameterized and tested with branches of black spruce (Picea mariana (Mill.) B.S.P.) and jack pine (Pinus banksiana Lamb.) trees growing in the Northern Study Area of the Boreal Ecosystem Atmosphere Study (BOREAS) in Manitoba, Canada. Branch samples containing foliage of all age-classes were harvested from a lowland old black spruce (OBS) and an old jack pine (OJP) stand and the responses of photosynthesis (A(n)) and stomatal conductance (g(s)) to temperature, CO(2), light, and leaf-to-air vapor pressure difference (VPD) were determined under controlled laboratory conditions at the beginning, middle, and end of the growing season (Intensive Field Campaigns (IFC) 1, 2, and 3, respectively). The parameterized model was then tested against in situ field gas-exchange measurements in a young jack pine (YJP) and an upland black spruce (UBS) stand as well as in the OBS and OJP stands. Parameterization showed that Rubisco capacity (V(max)), apparent quantum yield (alpha') and Q(10) for sink limitation were the most crucial parameters for the photosynthesis sub model and that V(max) varied among different measurement series in the laboratory. Verification of the model against the data used to parameterize it yielded correlation coefficients (r) of 0.97 and 0.93 for black spruce and jack pine, respectively, when IFC-specific parameters were used, and 0.77 and 0.87 when IFC-2 parameters were applied to all IFCs. For both measured and modeled g(s), the stomatal conductance sub-model, which linearly relates g(s) to (A(n)h(s))/c(s) (where h(s) and c(s) are relative humidity and CO(2) mole fraction at the leaf surface, respectively), had significantly steeper slopes and higher r values when only the VPD response data were used for parameterization than when all of the response data were used for parameterization. Testing the photosynthesis sub-model against upper canopy field data yielded poor results when laboratory estimates of V(max) were used. Use of the mean V(max) estimated for all upper canopy branches measured on a given day improved model performance for jack pine (from a nonsignificant correlation between measured and modeled A(n) to r = 0.45), but not for black spruce (r = 0.45 for both cases). However, when V(max) was estimated for each branch sample individually, the model accurately predicted the 23 to 137% diurnal variation in A(n) for all stands for both the upper and lower canopy. This was true both when all of the other parameters were IFC-specific (r = 0.93 and 0.92 for black spruce and jack pine, respectively) and when only mid-growing season (IFC-2) values were used (r = 0.92 for both species). Branch-specific V(max) estimates also permitted accurate prediction of field g(s) (r = 0.75 and 0.89 for black spruce and jack pine, respectively), although parameterization with all of the response data overestimated g(s) in the field, whereas parameterization with only the VPD response data provided unbiased predictions. Thus, after parameterization with the laboratory data, accurately modeling the range of A(n) and g(s) encountered in the field for both black spruce and jack pine was reduced to a single unknown parameter, V(max). PMID- 12651385 TI - Effect of nitrogen on the seasonal course of growth and maintenance respiration in stems of Norway spruce trees. AB - To determine effects of stem nitrogen concentration ([N]) on the seasonal course of respiration, rates of stem respiration of ten control and ten irrigated fertilized (IL), 30-year-old Norway spruce trees (Picea abies (L.) Karst.), growing in northern Sweden, were measured on seven occasions from June 1993 to April 1994. To explore sources of seasonal variation and mechanisms of fertilization effects on respiration, we separated total respiration into growth and maintenance respiration for both xylem and phloem bark. Stem respiration increased in response to the IL treatment and was positively correlated with growth rate, volume of living cells and stem nitrogen content. However, no significant effect of IL treatment or [N] in the living cells was found for respiration per unit volume of live cells. Total stem respiration during the growing season (June to September) was estimated to be 16.7 and 29.7 mol CO(2) m( 2) for control and IL-treated trees, respectively. Respiration during the growing season accounted for approximately 64% of total annual respiration. Depending on the method, estimated growth respiration varied between 40 and 60% of total respiration during the growing season. Between 75 and 80% of the live cell volume in the stems was in the phloem, and phloem maintenance accounted for about 70% of maintenance respiration. Because most of the living cells were found in the phloem, and the living xylem cells were concentrated in the outer growth rings, we concluded that the best base for expressing rates of stem growth and maintenance respiration in young Norway spruce trees is stem surface area. PMID- 12651386 TI - Physiological basis of the light use efficiency model. AB - The observation that, for unstressed plants, light use efficiency of a plant canopy, defined as the ratio of net primary productivity (NPP) to absorbed photosynthetically active radiation (APAR), is approximately constant with respect to changes in APAR, implies that NPP can be modeled using a linear relationship with APAR. However, such a linear relationship is counter-intuitive because the relationship between leaf photosynthesis and absorbed light is strongly nonlinear. Three arguments have been advanced to explain the observed linear relationship between NPP and APAR. In this paper, a detailed, physiologically based model of canopy radiation absorption and photosynthesis (MAESTRO) was used to analyze these arguments. The first argument is that the canopy is structured so that radiation is distributed throughout the canopy such that most leaves are exposed to non-saturating quantum flux density, resulting in a linear response of canopy photosynthesis to APAR. Simulations of MAESTRO indicated that this explanation is inadequate, because daily values of canopy photosynthetic light use efficiency calculated with MAESTRO were highly variable regardless of canopy structure. The second argument is that variability in light use efficiency decreases with increasing time scale. The simulations showed that this is true to some extent, although simulated annual canopy photosynthetic light use efficiency still varies across sites with different LAI or light climate. The third argument is that changes in canopy nitrogen content act both to maximize net canopy photosynthesis and to keep light use efficiency constant. This argument could not be tested with the model, but the failure of the first two explanations suggests that this third explanation deserves closer attention. PMID- 12651387 TI - Limitations of a compensation heat pulse velocity system at low sap flow: implications for measurements at night and in shaded trees. AB - Unlike an ideal system, the return time to thermal balance (t(b)) between upstream and downstream thermistors, as measured by the (compensation) heat pulse velocity method, effectively depends on the heat input and the water content of the wood at zero and low sap flow. Even when these factors were held constant and ambient temperature was stabilized, a twofold variation in t(b) at zero flow was observed within and among Greenspan Technology sensors implanted in wooden posts, making it impossible to distinguish zero flow from low sap velocities (< 0.01 0.02 mm s(-1)). This limitation has serious consequences because the contribution of low flow rates to water movement is important during both daytime and nighttime in tropical understory and overstory trees. Measurements in an artificial flow system showed that this technical limitation is exacerbated by erratic variation in sensor response at both zero and low flow rates. The limited sensitivity of the tested sap flow sensors may be caused by their poor thermal contact with wood. Interim procedures are suggested for estimating minimum detectable sap flow and delimiting the hydroactive zone until the sensitivity and interchangeability of sap flow probes are improved. PMID- 12651388 TI - Conversion of total leaf area to projected leaf area in lodgepole pine and Douglas-fir. AB - Three definitions of leaf area index (LAI) in the literature have no predictable relationship with each other. Factors were derived for converting total LAI to projected LAI of horizontal leaves and to projected LAI for inclined leaves of lodgepole pine (Pinus contorta Dougl. var. latifolia Engelm.) and coastal Douglas fir (Pseudotsuga menziesii (Mirb.) Franco) to allow comparison of results from different studies. An algorithm was derived to allow determination of these factors based on twig angles and the angles that the foliage subtends with the twig. Allowances were made for both the vertical and horizontal components of projection. The value of the factor for converting total LAI to projected LAI for inclined leaves was 0.229 + 0.0032 for lodgepole pine and 0.230 + 0.0037 for Douglas-fir. Sensitivity analysis established that this conversion factor was more sensitive to differences in vertical angles of the twigs than to twig rotation or foliar arrangement on the twig. PMID- 12651389 TI - Nutrient enrichment of white spruce seedlings during nursery culture and initial plantation establishment. AB - Growth and nutrient dynamics of bare-root white spruce (Picea glauca (Moench) Voss) seedlings were monitored for three years in a nursery and for one season after planting to assess effects of three nursery fertilization regimes. Nitrogen (totaling 0, 650, and 1300 kg N ha(-1) for 3 years) was applied conventionally in equal amounts during the growing season. The high-N treatment, representing a nutrient loading regime, was also applied seasonally at exponentially increasing rates as a fourth treatment. By the end of the 3-year nursery rotation, the N treatments had stimulated growth by 104-180% and nutrient uptake by as much as 381, 224 and 145% for N, P and K, respectively, inducing large accumulations of N and P in both conventionally and exponentially loaded seedlings. Compared with exponentially loaded seedlings, the concentrations of nutrients were less in conventionally loaded seedlings although their biomass was larger (31%). High nutrient reserves in the seedlings at the end of the nursery rotation resulted in increased biomass production (40-190%) after planting, which was related to the ability of the seedlings to retranslocate internal reserves for new growth, despite little or no net uptake of nutrients during the first season after planting. Compared with conventionally loaded seedlings, retranslocation was greater in exponentially loaded seedlings, which had accumulated larger and more readily available nutrient reserves during the nursery phase. The shortness of this study limits its usefulness for predicting the persistence of the loading response after planting, but we postulate that the high nutrient status of loaded seedlings at the end of the nursery rotation will contribute to future growth through increased nutrient storage and retranslocation, thus prolonging the loading response. PMID- 12651390 TI - Storage of foliar-absorbed nitrogen and remobilization for spring growth in young nectarine (Prunus persica var. nectarina) trees. AB - The effectiveness of spraying foliage with urea to provide nitrogen (N) to augment the seasonal internal cycling of N in young nectarine trees (Prunus persica (L.) Batsch var. nectarina (Ait. f. Maxim.), cv. Stark Red Gold) was studied. One-year-old trees were grown with contrasting N supplies during the summer and foliage was sprayed with a 2% urea solution labeled with (15)N just before leaf senescence started. After leaf abscission had finished, the trees were repotted in sand and given no further N. Remobilization of both labeled and unlabeled N for leaf growth the following spring was quantified. Leaves absorbed between 58 and 69% of the (15)N intercepted by the canopy irrespective of tree N status. During leaf senescence, the majority of (15)N was withdrawn from the leaves into the shoot and roots. Remobilization of (15)N the following spring was also unaffected by tree N status. About 38-46% of (15)N in the trees was recovered in the new growth. More unlabeled N (derived from root uptake) was remobilized for leaf growth in the spring than was withdrawn from leaves during canopy senescence the previous autumn. Therefore, soil-applied N augmented N storage pools directly, and contributed more to N remobilization the following spring than did foliar-absorbed (15)N. PMID- 12651391 TI - Whole-plant water flux in understory red maple exposed to altered precipitation regimes. AB - Sap flow gauges were used to estimate whole-plant water flux for five stem diameter classes of red maple (Acer rubrum L.) growing in the understory of an upland oak forest and exposed to one of three large-scale (0.64 ha) manipulations of soil water content. This Throughfall Displacement Experiment (TDE) used subcanopy troughs to intercept roughly 30% of the throughfall on a "dry" plot and a series of pipes to move this collected precipitation across an "ambient" plot and onto a "wet" plot. Saplings with a stem diameter larger than 10 cm lost water at rates 50-fold greater than saplings with a stem diameter of 1 to 2 cm (326 versus 6.4 mol H(2)O tree(-1) day(-1)). These size-class differences were driven largely by differences in leaf area and cross-sectional sapwood area, because rates of water flux expressed per unit leaf area (6.90 mol H(2)O m(-2) day(-1)) or sapwood area (288 mol H(2)O dm(-2) day(-1)) were similar among saplings of the five size classes. Daily and hourly rates of transpiration expressed per unit leaf area varied throughout much of the season, as did soil matrix potentials, and treatment differences due to the TDE were observed during two of the seven sampling periods. On July 6, midday rates of transpiration averaged 1.88 mol H(2)O m(-2) h(-1) for saplings in the "wet" plot, 1.22 mol H(2)O m(-2) h(-1) for saplings in the "ambient" plot, and 0.76 mol H(2)O m(-2) h(-1) for saplings in the "dry" plot. During the early afternoon of August 28, transpiration rates were sevenfold lower for saplings in the "dry" plot compared to saplings in the "wet" plot and 2.5-fold lower compared to saplings in the "ambient" plot. Treatment differences in crown conductance followed a pattern similar to that of transpiration, with values that averaged 60% lower for saplings in the "dry" plot compared to saplings in the "wet" plot and 35% lower compared to saplings in the "ambient" plot. Stomatal and boundary layer conductances were roughly equal in magnitude. Estimates of the decoupling coefficient (Omega) ranged between 0.64 and 0.72 for saplings in the three TDE treatment plots. We conclude that red maple saplings growing in the understory of an upland oak forest are responsive to their edaphic and climatic surroundings, and because of either their small stature or their shallow root distribution, or both, are likely to be impacted by precipitation changes similar to those predicted by global climate models. PMID- 12651392 TI - Ozone-induced changes in biosynthesis of Rubisco and associated compensation to stress in foliage of hybrid poplar. AB - Experiments were conducted during the growing seasons of 1993-1995 to determine whether exposure to ozone (O(3)) affected the synthesis of ribulose-1,5 bisphosphate carboxylase/oxygenase (Rubisco) in hybrid poplar, Populus maximowizii A. Henry x trichocarpa Torr. & A. Gray, Clone 245. As the canopy aged, the concentration of Rubisco decreased at a more rapid rate in lower leaves of hyrid poplar ramets subjected to chronic O(3) exposure in open-top chambers than in comparable foliage of plants grown in charcoal-filtered air. There was no difference in rate of synthesis of Rubisco between treatments, suggesting that loss of this protein in O(3)-treated leaves was caused by an accelerated rate of proteolysis. In foliage higher in the canopy, both concentration and rate of synthesis of Rubisco were stimulated by O(3) for a brief period when the leaves were young. Quantification of mRNA for the small (rbcS) and large (rbcL) transcripts of Rubisco did not reveal changes that were likely to reflect altered synthesis of Rubisco as a prime response to O(3). Analyses of Rubisco concentration and rate of Rubisco synthesis in foliage connected by vascular traces within the canopy indicated that loss of Rubisco in older leaves was associated with an increase in this protein in younger leaves higher in the canopy. These data support the notion that accelerated senescence may provide some compensatory benefit to the plant. In 1995, the rate of synthesis of Rubisco was almost always higher in O(3)-treated foliage than in nontreated foliage, even when the concentration of Rubisco was adversely affected by the O(3) treatment. Because accelerated foliar abscission in response to O(3) was minimal in 1995 compared to other years, we speculate that, when abscission is delayed, Rubisco synthesis and concentration become uncoupled. PMID- 12651393 TI - A model of dry matter partitioning in trees. AB - In mechanistic growth models, the description of assimilate allocation or dry matter partitioning plays a key role. Although theoretical concepts of allocation exist, they include many parameters that cannot be quantified. Therefore, many growth models use descriptive keys that represent the proportions of dry matter or carbohydrates assigned to each plant component. I have developed a model to describe the dynamic partitioning of dry matter in individual trees, and used it to investigate the effects of growth conditions on the partitioning pattern in Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) and beech (Fagus sylvatica L.). The model estimates the fractions of total available dry matter that should go to certain plant parts, based on the concept of structural balances. Both mechanistic and allometric relationships between tree components are used to model conditions for the dynamic distribution of dry matter. The model was to used to estimate the effects of dominance position, site conditions, and thinning on growth partitioning. The fractions of the annual current increment of total dry matter gradually changed with tree age, but the changes were relatively small, especially after age 20. Compared with beech, Douglas-fir invested more dry matter in foliage, especially at the cost of the branch and stem components. Trees of average basal area invested more dry matter in branches and less in stem than suppressed trees, and their estimated increase in stem diameter over time generally fitted the yield table data well. Stem diameter development was underestimated at higher ages only in the case of a Douglas-fir tree of average basal area on a poor site. Over time, the proportion of standing biomass in foliage and fine root fractions showed a gradual decline, whereas there was a gradual increase in the proportion of standing biomass in the stem fraction. These age-related changes were attributed to different loss rates among components. Analysis of the effects of thinning revealed that a discontinuous reduction in stem number results in a slow decrease in partitioning to the stem. The most obvious response to thinning consisted in a sharp decrease in partitioning to fine roots and foliage, and an increased investment in branches. Stem diameter growth appeared relatively constant in response to thinning, indicating that it will increase almost linearly with time. I conclude that the model is able to reproduce the development of an individual tree over time, both in terms of stem diameter and biomass. The model is thus suitable for simulating the effects of competition for resources on growth and development of forest stands. PMID- 12651394 TI - In vitro effects of Laccaria bicolor S238 N and Pseudomonas fluorescens strain BBc6 on rooting of de-rooted shoot hypocotyls of Norway spruce. AB - The ectomycorrhizal fungus Laccaria bicolor S238 N and the bacterium Pseudomonas fluorescens BBc6 were used separately and in combination to induce in vitro rooting of de-rooted shoot hypocotyls of Norway spruce (Picea abies (L.) Karst.). When the culture medium was supplemented with tryptophan, a precursor of indole-3 acetic acid (IAA) synthesis, the presence of the ectomycorrhizal fungus increased the percentage of hypocotyls forming roots; furthermore, both the fungal and bacterial inoculations enhanced the number of roots formed per rooted hypocotyl. Similar results were obtained by adding exogenous IAA (5 and 10 &mgr;M) to the rooting medium. After the rooting phase, the fungal inoculation enhanced adventitious root elongation and branching as well as the aerial growth of the cuttings. Pseudomonas fluorescens BBc6 had no effect on these parameters. The production of IAA by pure cultures of L. bicolor S238 N and P. fluorescens BBc6 was estimated by immunochemical analysis using specific anti-IAA antibodies. Both L. bicolor S238 N and P. fluorescens BBc6 synthesized IAA in pure culture and synthesis was stimulated in the presence of tryptophan. Thus, the effect of the fungus in stimulating adventitious root formation and subsequent elongation and branching can be attributed, at least partially, to the synthesis of IAA by the fungus. The finding that P. fluorescens BBc6 had no effect on root elongation and branching although it produced IAA suggests that either IAA was not the only parameter involved in the stimulation of these processes by L. bicolor S238 N or the bacterium produced other compounds that counteracted the stimulatory effects of IAA on root elongation and branching. PMID- 12651395 TI - Responses of citrus fine roots to localized soil drying: a comparison of seedlings with adult fruiting trees. AB - We studied the responses of citrus (Citrus volkameriana Tan. & Pasq.) roots to 15 weeks of soil drying. A comparison was made between the fine roots of 1-year-old seedling root systems (seedling) and the fine roots of woody laterals of 6-year old grafted trees (adult). Each seedling and woody lateral root system was established in a pair of vertically separated and independently irrigated soil compartments located in field root chambers excavated adjacent to the trees to which the woody laterals were attached. Root + soil respiration and fine root survival of seedlings and adults were similar for the first 5 weeks. However, eight weeks after termination of irrigation to the upper soil compartments, mortality of fine roots was high in adults but not seedlings. Fine roots of adults exposed to dry soil for 5, 8 and 15 weeks exhibited 2, 26 and 33% mortality, respectively, whereas the corresponding values for fine roots of seedlings were 2, 6 and 8%. Although root + soil respiration rates of adults and seedlings were similar before the soil drying treatment, rates for adults were only 25% of those for seedlings after 15 weeks of soil drying. We conclude that, although fine roots of adults and seedlings are similar in form, they respond differently to soil drying. PMID- 12651396 TI - Paclobutrazol affects the resistance of black spruce to high light and thermal stress. AB - Detached needles from 20-week-old black spruce (Picea mariana (Mill.) B.S.P.) seedlings root-drenched with 60 mg of paclobutrazol were exposed to two temperatures (22 and 50 degrees C) and two light treatments (100 and 1900 &mgr;mol m(-2) s(-1) PAR) in a factorial combination for 4 h in vitro. Mean dry weights of individual needles from paclobutrazol-treated plants were approximately 1.9 times heavier than that of needles from untreated controls at 22 degrees C, but no differences were observed following incubation at 50 degrees C. Numbers of cells per needle remained constant in all treatments. Chlorophyll and carotenoid contents per needle were higher in seedlings treated with paclobutrazol than in untreated control seedlings, and the differences were most pronounced in the high temperature plus high light treatment. In low light at 50 degrees C, quantum efficiency of photosystem II was 45% higher in needles of paclobutrazol-treated seedlings than in needles of untreated control seedlings, but quantum efficiency of needles from treated seedlings declined when needles were exposed to high light at either temperature. Peroxidase and superoxide dismutase activities were up-regulated by paclobutrazol, whereas catalase activities were depressed and no significant differences were observed between treated and control needles at 50 degrees C in either light treatment. Paclobutrazol treatment did not moderate the depressive effects of high temperature on total soluble protein or on the activity of ribulose-1,5 bisphosphate carboxylase. In contrast, high activities of phosphoenolpyruvate carboxylase were maintained in paclobutrazol-treated needles under all stress conditions, whereas large losses in activity were recorded in untreated needles at 50 degrees C. Collectively, these observations suggest that paclobutrazol treatment may convey resistance to excessive light and high temperatures by increasing the potential of conifers to limit damage caused by oxidative stress. PMID- 12651397 TI - Net primary production of forests: a constant fraction of gross primary production? AB - Considerable progress has been made in our ability to model and measure annual gross primary production (GPP) by terrestrial vegetation. But challenges remain in estimating maintenance respiration (R(m)) and net primary production (NPP). To search for possible common relationships, we assembled annual carbon budgets from six evergreen and one deciduous forest in Oregon, USA, three pine plantations in New South Wales, Australia, a deciduous forest in Massachusetts, USA, and a Nothofagus forest on the South Island of New Zealand. At all 12 sites, a standard procedure was followed to estimate annual NPP of foliage, branches, stems, and roots, the carbon expended in synthesis of these organs (R(g)), their R(m), and that of previously produced foliage and sapwood in boles, branches, and large roots. In the survey, total NPP ranged from 120 to 1660 g C m(-2) year(-1), whereas the calculated fraction allocated to roots varied from 0.22 to 0.63. Comparative analysis indicated that the total NPP/GPP ratio was conservative (0.47 +/- 0.04 SD). This finding supports the possibility of greatly simplifying forest growth models. The constancy of the NPP/GPP ratio also provides an incentive to renew efforts to understand the environmental factors affecting partitioning of NPP above and belowground. PMID- 12651398 TI - Application of chlorophyll fluorescence to evaluate Mn tolerance of deciduous broad-leaved tree seedlings native to northern Japan. AB - We used chlorophyll fluorescence to examine photosynthetic responses to excess Mn accumulation in leaves of four tree species differing in successional traits. Betula ermanii Cham. (Be) and Alnus hirsuta Turcz. (Ah) were studied as representatives of early-successional species. Ulmus davidiana Planch. var. japonica (Rehder) Nakai (Ud) was selected as a mid-successional species, and Acer mono Maxim. var. glabrum (Lev. et Van't.) Hara (Am) was chosen as a late successional species. In Be, Ah and Am, high foliar concentrations of Mn had little effect on maximum photochemical efficiency of photosystem II (PSII), as indicated by the values of dark-adapted F(v)/F(m), whereas a significant decrease was observed in Ud. Photochemical quenching (qP) and the excitation capture efficiency of open PSII (F'(v)/F'(m)) decreased with increasing leaf Mn concentration at photosynthetic steady state after a 15-min exposure to 430 &mgr;mol m(-2) s(-1) PPFD. Compared with early-successional species, these decreases were greater in mid- and late-successional species leading to lower effective quantum efficiencies of PSII (DeltaF/F'(m) = qP x F'(v)/F'(m) = (F'(m) F)/F'(m)). To determine the extent of photoinhibition, F(v)/F(m) of the illuminated leaves was remeasured after a 15-min dark period. Compared with the dark-adapted F(v)/F(m), we observed a significant decrease in F(v)/F(m) in Am leaves containing high concentrations of Mn. These chlorophyll fluorescence studies indicate that the early-successional species Be and Ah have a higher tolerance to excessive accumulations of Mn in leaves than the mid- and late successional species Ud and Am. PMID- 12651399 TI - Net ecosystem productivity, net primary productivity and ecosystem carbon sequestration in a Pinus radiata plantation subject to soil water deficit. AB - Tree carbon (C) uptake (net primary productivity excluding fine root turnover, NPP') in a New Zealand Pinus radiata D. Don plantation (42 degrees 52' S, 172 degrees 45' E) growing in a region subject to summer soil water deficit was investigated jointly with canopy assimilation (A(c)) and ecosystem-atmosphere C exchange rate (net ecosystem productivity, NEP). Net primary productivity was derived from biweekly stem diameter growth measurements using allometric relations, established after selective tree harvesting, and a litterfall model. Estimates of A(c) and NEP were used to drive a biochemically based and environmentally constrained model validated by seasonal eddy covariance measurements. Over three years with variable rainfall, NPP' varied between 8.8 and 10.6 Mg C ha(-1) year(-1), whereas A(c) and NEP were 16.9 to 18.4 Mg C ha(-1) year(-1) and 5.0-7.2 Mg C ha(-1) year(-1), respectively. At the end of the growing season, C was mostly allocated to wood, with nearly half (47%) to stems and 27% to coarse roots. On an annual basis, the ratio of NEP to stand stem volume growth rate was 0.24 +/- 0.02 Mg C m(-3). The conservative nature of this ratio suggests that annual NEP can be estimated from forest yield tables. On a biweekly basis, NPP' repeatedly lagged A(c), suggesting the occurrence of intermediate C storage. Seasonal NPP'/A(c) thus varied between nearly zero and one. On an annual basis, however, NPP'/A(c) was 0.54 +/- 0.03, indicating a conservative allocation of C to autotrophic respiration. In the water-limited environment, variation in C sequestration rate was largely accounted for by a parameter integrative for changes in soil water content. The combination of mensurational data with canopy and ecosystem C fluxes yielded an estimate of heterotrophic respiration (NPP' - NEP) approximately 30% of NPP' and approximately 50% of NEP. The estimation of fine-root turnover rate is discussed. PMID- 12651400 TI - Quantitative estimates of uptake and internal cycling of (14)N-labeled fertilizer in mature walnut trees. AB - Uptake and internal cycling of nitrogen (N) in mature walnut trees was studied over a period of 6 years using (15)N-depleted fertilizer and full-canopied walnut (Juglans regia L. cv Hartley) trees. The magnitude of internal N cycling, i.e., the availability of N for new growth from internal N pools, was quantified using both the percent annual depletion (PAD) and the N balance budget approaches. There was good agreement between the two measures, and about 60% of annual N demand was derived from N redistribution from internal pools. The remaining 40% of annual tree N demand was met by an influx of N from the soil/fertilizer pool. Trees were excavated, processed and analyzed after 6 years to determine total tree N content and labeled N recovery. Trees recovered 29.4% of the labeled N applied and, based on previous evidence, we assumed that tree accumulation of labeled N occurred entirely in the first year. Labeled N in the fruits and leaves harvested in the first year represented 26% of the total labeled N accumulated, and the remaining 74% of the labeled N accumulated that year was stored and used to support development of annual organs in subsequent years. In the first year, the early maturing catkins did not accumulate labeled N, indicating their exclusive reliance on internal N. Using the atom% (14)N excess values of catkins and an exponential decline equation to determine turnover rate, the Mean Residence Time (MRT) of storage N in the tree was estimated to be 2.0 years. The size of the cycling pool of storage N in the tree was estimated to be about 50% of the total N content of perennial tree parts. Our data support the hypotheses that: (1) in any given year, mature walnut trees store the majority of soil and fertilizer N absorbed and within 2 years following uptake the N is remobilized and used for new growth, and (2) about half of the total N content of the perennial parts of mature walnut trees is present as nonstructural N and is available for recycling. PMID- 12651401 TI - Biomass and nutrient allocation in Douglas-fir and amabilis fir seedlings: influence of growth rate and nutrition. AB - Allocation of biomass and nutrients to shoots and roots was followed for three years in fast and slow growing populations of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco), a fast growing pioneer species, and amabilis fir (Abies amabilis Dougl. ex J. Forbes), a slow growing shade-tolerant species. Seedlings were grown for three seasons in five nutrient treatments containing varying proportions of nitrogen and phosphorus (N:P). In both species, growth was greatest in the 250:20 N:P treatment followed by the 100:60 and 100:20 treatments. Vector analysis showed that, in both species, relative to the 100:20 treatment, seedlings in the 20:20 treatment were N deficient and seedlings in the 100:4 treatment were P deficient, i.e., where deficiency is defined to mean that an increase in nutrient supply increases nutrient content, nutrient concentration and plant dry weight. Seedlings in the 100:60 treatment had a higher P content than seedlings in the 100:20 treatment but the same dry weight, indicative of what Timmer and Armstrong (1987) termed luxury consumption. No nutrient retranslocation was observed in either species until the third growing season. In Douglas-fir, the greatest percentage of nutrients was exported from one-year-old shoots between May and July of the third growing season to support new growth. The total amount and percent of nutrients retranslocated was higher in Douglas-fir than in amabilis fir. Amabilis fir seedlings also exported N and P from older shoots, but this was later partially replenished. In both species, P retranslocation was greatest in treatments with a high N:P ratio. Nitrogen retranslocation was greatest in amabilis fir seedlings in treatments with a low N:P ratio, and greatest in Douglas-fir seedlings in the 250:20 and 100:60 treatments. Potassium retranslocation was correlated with seedling size. Douglas-fir retranslocated more of its shoot N reserves into new growth at the expense of older needles when soil fertility was high and sinks were strong. Otherwise, both species preferentially translocated the elements in short supply. Thus, retranslocation varied with the ecological characteristics of species, the relative availability of soil nutrients and sink strength. PMID- 12651402 TI - Effects of photoperiod and temperature on the timing of bud burst in Norway spruce (Picea abies). AB - We examined the effects of several photoperiod and temperature regimes imposed during the winter-spring period on the timing of bud burst in rooted cuttings of Norway spruce (Picea abies (L.) Karst.) grown in a greenhouse in Finland. The treatments were initiated in November and December after the cuttings had been exposed to natural chilling and freezing events. Irrespective of the treatments applied, time to bud burst decreased with increased duration of previous exposure to natural chilling and freezing events. Fluctuating day/night temperatures and continuous lengthening of the photoperiod hastened bud burst. Shortening the photoperiod delayed bud burst, suggesting that little or no ontogenetic development toward bud burst takes place during mild periods before the winter solstice. In the case of climatic warming, this phenomenon may prevent the premature onset of growth that has been predicted by computer simulations with models that only consider temperature regulation of bud burst. PMID- 12651403 TI - Flower induction treatments have no effects on seed traits and transmission of alleles in Picea glauca. AB - Flower induction methods-hormone application or exposure to physiological stress, or both-are used routinely for shortening breeding cycles and increasing seed production in white spruce (Picea glauca (Moench) Voss). The objectives of this study were: (1) to evaluate the effects of flower induction on seed yield and quality in white spruce; and (2) to determine if flower induction treatments affect the maternal contribution to offspring. We assessed the effects of flower induction treatments, which consisted of gibberellin A(4/7) (GA(4/7)) and naphthaleneacetic acid (NAA) stem injections, on allele segregation for 28 clones, number of seeds per cone, number of sound seeds per cone, seed weight, and the germination rate of a subset of clones. Flower induction treatments did not affect any of the phenotypic traits examined. No increase in segregation distortion in allozyme loci following flower induction treatments was observed. PMID- 12651404 TI - Nitrogen-related root variables of trees along an N-deposition gradient in Europe. AB - We measured fine root N concentration, root in vivo nitrate reductase activity (NRA) and root uptake capacity for (15)NH(4) (+) and (15)NO(3) (-) along an N deposition gradient from northern Sweden to central Europe, encompassing a variation in N deposition rates of < 5 to about 40 kg N ha(-1) year(-1). The focus was on Picea abies (L.) Karst., but Fagus sylvatica L. in central Europe and Pinus sylvestris L. and Betula spp. in northern Sweden were also studied. We assumed that, with an increased supply of N, root N concentration would increase, activity of the inducible enzyme nitrate reductase (NR) in roots would increase, particularly with an increasing supply of NO(3) (-), and root uptake capacity for inorganic N would decline, reflecting a lower demand for N. As expected, fine root N concentration in P. abies increased along the gradient from 1.1% (d.w. basis) at the northern site to 2.1% at central European sites. This variation compared with an amplitude of 0.7-1.5% for foliage. Root in vivo NRA was low in northern Sweden, and higher in central Europe. Picea abies and broad-leaved species had similar root NRA. At one location in Denmark and one in France, however, root NRA in the spring was very high in F. sylvatica. Root uptake capacity for NO(3) (-), as measured in excised roots, was low throughout the transect, but in P. abies, it was high for NH(4) (+) in northern Sweden and decreased by a factor of 4 with increasing N deposition. A similar pattern was found in the broad-leaved species. Unless the higher availability of NO(3) (-) and lower specific root uptake capacity per unit root mass for inorganic N in central Europe (compared with northern Sweden) is balanced by a higher root biomass, the central European forests will be a weaker sink for N. PMID- 12651405 TI - Growth, productivity and water use in three hybrid poplar clones. AB - Three drought treatments (none, medium and severe) were applied to three container-grown, hybrid poplar clones (Beaupre, Trichobel and Ghoy) as a series of drying cycles over two years-in a greenhouse in 1994 and outside in 1995. Leaf area development, annual biomass production and annual water use were recorded. Clone Beaupre had greater rates of leaf area development (up to 11.8 x 10(-4) m(2) degrees C day(-1)) and subsequently greater dry matter production (up to 816 g tree(-1) year(-1)) and water use (up to 168 l tree(-1) year(-1)) than the other two clones. In all clones, rates of leaf area development, annual biomass production and water use were less under drought conditions than under well watered conditions. On a proportional basis, the highest reduction in biomass by drought was in Trichobel (up to 74%), whereas, in absolute terms, the reduction was greatest in Beaupre (up to 500 g tree(-1)). However, under drought conditions, annual biomass production was greater in Clone Beaupre than in the other clones. The relationship between water use and biomass production was constant over both years, and was similar for Beaupre and Trichobel at 4.4 g l( 1), but was less for Ghoy at 3.5 g l(-1). PMID- 12651406 TI - Seedlings of five boreal tree species differ in acclimation of net photosynthesis to elevated CO(2) and temperature. AB - Biochemical models of photosynthesis suggest that rising temperatures will increase rates of net carbon dioxide assimilation and enhance plant responses to increasing atmospheric concentrations of CO(2). We tested this hypothesis by evaluating acclimation and ontogenetic drift in net photosynthesis in seedlings of five boreal tree species grown at 370 and 580 &mgr;mol mol(-1) CO(2) in combination with day/night temperatures of 18/12, 21/15, 24/18, 27/21, and 30/24 degrees C. Leaf-area-based rates of net photosynthesis increased between 13 and 36% among species in plants grown and measured in elevated CO(2) compared to ambient CO(2). These CO(2)-induced increases in net photosynthesis were greater for slower-growing Picea mariana (Mill.) B.S.P., Pinus banksiana Lamb., and Larix laricina (Du Roi) K. Koch than for faster-growing Populus tremuloides Michx. and Betula papyrifera Marsh., paralleling longer-term growth differences between CO(2) treatments. Measures at common CO(2) concentrations revealed that net photosynthesis was down-regulated in plants grown at elevated CO(2). In situ leaf gas exchange rates varied minimally across temperature treatments and, contrary to predictions, increasing growth temperatures did not enhance the response of net photosynthesis to elevated CO(2) in four of the five species. Overall, the species exhibited declines in specific leaf area and leaf nitrogen concentration, and increases in total nonstructural carbohydrates in response to CO(2) enrichment. Consequently, the elevated CO(2) treatment enhanced rates of net photosynthesis much more when expressed on a leaf area basis (25%) than when expressed on a leaf mass basis (10%). In all species, rates of leaf net CO(2) exchange exhibited modest declines with increasing plant size through ontogeny. Among the conifers, enhancements of photosynthetic rates in elevated CO(2) were sustained through time across a wide range of plant sizes. In contrast, for Populus tremuloides and B. papyrifera, mass-based photosynthetic rates did not differ between CO(2) treatments. Overall, net photosynthetic rates were highly correlated with relative growth rate as it varied among species and treatment combinations through time. We conclude that interspecific variation may be a more important determinant of photosynthetic response to CO(2) than temperature. PMID- 12651407 TI - Leaf distribution in large trees and stands of the floodplain forest in southern Moravia. AB - Vertical distributions of leaf dry mass (M(d)) and leaf area (A(f)) were related to relative irradiance (I(r); I(r) above the stand = 1) in closed-canopy, old growth stands of the floodplain forest in southern Moravia composed largely of Quercus, Fraxinus and Tilia species. Foliage area and mass at any given canopy height were converted to solar equivalent leaf area (A(s)) and mass (M(s)) by multiplying actual values at a given level in the canopy by the relative irradiance at that position. Stand leaf area index (LAI) was 5 (7 including shrub and herb layer), and solar equivalent parameters reached about 25% of that amount. In all species, vertical profiles of both relative irradiance and leaf dry mass to area ratio (LMA) were sigmoidal and the two variables were linearly related. The dominant, upper canopy species had a larger proportion of solar equivalent foliage than suppressed understory species. For individual trees of all species, the upper canopy had a larger proportion of solar equivalent foliage than the lower canopy. Light compensation points at both the leaf and whole-tree level were defined according to leaf or tree position, size and structure. I conclude that optimization of A(s) for forest stands may be used as a basis for determining thinning schedules and evaluating tree survival after damage to tree crowns by various factors. PMID- 12651408 TI - Influence of ectomycorrhizal fungal inoculation on growth and root IAA concentrations of transplanted conifers. AB - We determined whether in vitro plant growth regulator production by mycorrhizal fungi is correlated with conifer seedling growth and root IAA concentrations. Container-grown seedlings of interior Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco), lodgepole pine (Pinus contorta Dougl.) and ponderosa pine (Pinus ponderosa Dougl.) were inoculated at seeding with ectomycorrhizal fungi having a high, moderate or low capacity to produce either IAA or ethylene in vitro. Inoculated seedlings were grown for one season in the nursery, harvested in December, cold stored over winter and then transplanted to either a nursery field or a forest site in the spring. Seedling morphology and endogenous IAA in roots were measured immediately after cold storage and again six and 12 months after transplanting. Morphological responses to inoculation varied among different mycorrhizal fungi. Free IAA concentration of roots was increased in some inoculation treatments for all conifer species. In seedlings transplanted to a nursery field, in vitro ethylene-producing capacity of the ectomycorrhizal fungi was highly correlated with more morphological features than in vitro IAA producing capacity. Both IAA- and ethylene-producing capacity were significantly correlated with more morphological features in seedlings transplanted to a forest site than in seedings transplanted to a nursery field. One year after transplanting, only in vitro IAA-producing capacity was correlated with endogenous IAA concentration of roots of the inoculated seedlings. We conclude that growth responses of conifer seedlings can be partially influenced by IAA and ethylene produced by ectomycorrhizal fungal symbionts. PMID- 12651409 TI - Acclimation of Pacific yew (Taxus brevifolia) foliage to sun and shade. AB - The success in clinical trials of the anti-cancer drug, Taxol(R), obtained from the bark of Pacific yew (Taxus brevifolia Nutt.), has raised interest in cultivation and regeneration of this little-known species. Pacific yew is shade tolerant and it is not known whether the foliage can tolerate the high solar irradiances found on an open forest regeneration site or a nursery. Acclimation of Pacific yew to sun and shade was studied by comparing foliar physiology and morphology of male and female trees growing in full sun or shade. Interspecific foliar acclimation to sun was studied by comparing sun-grown English yew (Taxus baccata L.) with Pacific yew. No sex-specific acclimation was found in foliar physiology or morphology in either species. Sun-grown foliage of Pacific yew and English yew differed with respect to light harvesting, transpiration, stomatal conductance, leaf structure, stomatal distribution and foliar N concentrations and contents. Chlorophyll a fluorescence measurements indicated that shade-grown foliage of Pacific yew had larger and more efficient light harvesting systems than sun-grown foliage. Rates of CO(2) uptake and transpiration were similar in sun- and shade-grown foliage indicating acclimation of photosynthesis to the growth irradiance. Specific leaf area was significantly higher in shade-grown foliage of Pacific yew than in sun-grown foliage and was diagnostic of the light environment in which the foliage grew. Foliar N concentrations were not significantly different between sun- and shade-grown leaves of Pacific yew but sun-grown foliage had a higher N content. Physiological and morphological adjustments of Pacific yew foliage conferred tolerance to both high light and shade, enabling the trees to survive in a variety of light environments and indicating that Pacific yew is suited to nursery cultivation and regeneration of open sites. PMID- 12651410 TI - Shoot structure, light interception, and distribution of nitrogen in an Abies amabilis canopy. AB - We studied the effects of variation in shoot structure and needle morphology on the distributions of light and nitrogen within a Pacific silver fir (Abies amabilis (Dougl.) Forbes) canopy. Specifically, we investigated the role of morphological shade acclimation in the determination of resource use efficiency, which is claimed to be optimal when the distribution of nitrogen within the canopy is directly proportional to the distribution of intercepted photosynthetically active radiation (PAR). Shoots were collected from different heights in the crowns of trees representing four different size classes. A new method was developed to estimate seasonal light interceptance (SLI, intercepted PAR per unit needle area) of the shoots using a model for the directional distribution of above-canopy PAR, measurements of shoot silhouette area and canopy gap fraction in different directions. The ratio SLI/SLI(o), where the reference value SLI(o) represents the seasonal light interceptance of a spherical surface at the shoot location, was used to quantify the efficiency of light capture by a shoot. The ratio SLI/SLI(o) doubled from the top to the bottom of the canopy, mainly as a result of smaller internal shading in shade shoots than in sun shoots. Increased light-capturing efficiency of shade shoots implies that the difference in intercepted light by sun shoots versus shade shoots is much less than the decrease in available light from the upper to the lower canopy. For example, SLI of the five most sunlit shoots was only about 20 times greater than the SLI of the five most shaded shoots, whereas SLI(o) was 40 times greater for sun shoots than for shade shoots. Nitrogen content per unit needle area was about three times higher in sun needles than in shade needles. This variation, however, was not enough to produce proportionality between the amounts of nitrogen and intercepted PAR throughout the canopy. PMID- 12651411 TI - Stomatal conductance, growth and root signaling in Betula pendula seedlings subjected to partial soil drying. AB - Seedlings of Betula pendula Roth were grown with their root systems separated between two soil compartments. Four treatments were imposed: (i) adequate irrigation in both compartments (WW, controls); (ii) adequate irrigation in one compartment and drought in the other compartment (WD); (iii) drought in both compartments (DD); and (iv) half of the root system severed and the remainder kept well-watered (root excision, RE). Predawn leaf water potential, stomatal conductance, soil-to-leaf specific hydraulic conductance, and root and leaf growth decreased in DD-treated seedlings, which also displayed severe leaf shedding (30% loss in leaf area). The DD treatment also resulted in increased concentrations of abscisic acid (ABA) and its glucose ester in the xylem sap of roots and shoots compared to concentrations in control seedlings (about 200 versus 20 nM). Despite the difference in xylem sap concentrations, total ABA flux to the shoots was similar in the two treatments (1-2 pmol ABA m(-2) leaf area s( 1)) as a result of reduced transpiration in the DD-treated seedlings. Compared with root growth in control plants, root growth increased in the RE-treated plants and decreased in the drying compartment of the WD treatment; however, the RE and WD treatments only slightly reduced leaf expansion, and had no detectable effects on shoot water relations or ABA concentrations of the root and shoot xylem sap. We conclude that short-term soil water depletion affecting only 50% of the root system does not cause a measurable stress response in birch shoots, despite root growth cessation in the fraction of drying soil. PMID- 12651412 TI - Hydraulic properties of Douglas-fir (Pseudotsuga menziesii) branches and branch halves with reference to compression wood. AB - Douglas-fir (Pseudotsuga menziesii var. menziesii (Mirb.) Franco) branch segments were used to test the hypothesis that compression wood reduces xylem transport efficiency. Whole 3-year-old segments were first measured for specific conductivity (k(s), m(2) s(-1) MPa(-1)), then split lengthwise into upper and lower halves, the latter containing all or most of the compression wood in the segment. Halves were then remeasured for k(s) using a new technique that prevents leakage of permeating fluid during measurements. Lower branch halves had significantly lower k(s) than upper halves (6.4 +/- 0.3 versus 9.3 +/- 0.3 m(2) s(-1) MPa(-1) x 10(-4), respectively; n = 36), and despite their larger size, significantly lower hydraulic conductivity (k(h), m(4) s(-1) MPa(-1)) than upper halves. Lower branch halves had higher specific gravity (0.51 +/- 0.01 versus 0.45 +/- 0.01; n = 36), lower water content (123 +/- 2% versus 155 +/- 3%; n = 36), and larger proportions of volume occupied by both cell wall and air than upper halves. Lower halves had more tracheids per annual ring than upper halves (73 +/- 3 versus 63 +/- 2 per radial transect, respectively; n = 36), but tracheids were shorter and had narrower lumens than those of upper branch halves. Differences in hydraulic properties between upper and lower halves suggest that compression wood does reduce xylem transport efficiency. In contrast, the amount of compression wood in each sample did not explain any variation in whole unsplit sample hydraulic properties. PMID- 12651413 TI - Clonal and seasonal differences in leaf osmotic potential and organic solutes of five hybrid poplar clones grown under field conditions. AB - Leaf osmotic potential at full turgor (Psi(pio)) and the major solutes that contribute to osmotic potential were characterized in five hybrid poplar clones of Populus trichocarpa Torr. & Gray x P. deltoides Bartr. (TD) and P. deltoides x P. nigra L. (DN), growing under field conditions at two sites in eastern Washington and Oregon, USA. Trees were drip irrigated with 46, 76 or 137 cm of supplemental irrigation during each growing season. Trees at Wallula, WA, which were in their third growing season in 1994, were sampled twice a year for two years (1994 and 1995), and trees at Boardman, OR, which were in their second growing season in 1994, were sampled once a year for three years (1994-1996). At Wallula, the TD and DN clones exhibited lower predawn leaf water potentials in the 46-cm treatment than in the 137-cm treatment (-1.2 versus -0.7 MPa) during a hot, dry period in July 1994. Clone TD had a lower Psi(pio) than Clone DN (-1.67 versus -1.56 MPa) during the same period and the difference was also evident in 1995 (-1.81 versus -1.72 MPa) when trees were in their fourth growing season. There was also a significant treatment effect on Psi(pio) in Clone TD, with trees in the 46-cm treatment having lower Psi(pio) than trees in the 137-cm treatment in July 1994. At Boardman, Psi(pio) was generally high with no treatment differences during the 1994-96 samplings. The TD clones had significantly lower Psi(pio) than the DN clones in 1994 (-1.44 versus -1.36 MPa) and 1996 (-1.72 versus -1.54 MPa), but there was no difference between clones in 1995 (-1.40 versus -1.43 MPa). In 1995, at Wallula, osmotic adjustment in Clone TD was largely accounted for by an increase in sucrose, which constituted 70% of total organic solutes. Although the total concentration of free primary amino acids in this clone was 28% higher in trees in the 46-cm treatment than in trees in the 137-cm treatment, amino acids constituted only a small fraction of the total solute pool. Sixty-two percent of total solutes were inorganic ions in Clone TD compared to 52% in Clone DN, and potassium was the main ion constituting about 30% of total solutes and 50% of total ions. However, the clonal difference in Psi(pio) was not fully accounted for by the difference in solute concentration. Osmotic potential at full turgor declined over the growing season and with age. We conclude that, because the extent of osmotic adjustment exhibited by these clones was small, other drought resistance mechanisms contributed to the clonal differences in field performance. PMID- 12651414 TI - Drought resistance of two hybrid Populus clones grown in a large-scale plantation. AB - Poplar hybrids were grown with irrigation in a large-scale plantation to investigate the mechanisms underlying clonal differences in drought resistance. Beginning in spring 1992, Populus trichocarpa x P. deltoides (TD) and P. deltoides x P. nigra (DN) cuttings received 46, 76, or 137 cm year(-1) of irrigation to supplement the 18-20 cm of annual precipitation, and all trees received the same fertilization regime. Stem volume, assessed as the square of stem diameter at breast height times tree height (D(2)H), and water relations of the trees were studied from the end of their second growing season until the end of their fifth growing season. By the end of the second growing season, stem volume of Clone TD was 40-146% larger than that of Clone DN, but stem volume growth was independent of irrigation in excess of 46 cm year(-1) in both clones. During the third growing season, stem volume growth of both clones was limited by both the 46- and 76-cm irrigation treatments, so that by the end of the third growing season trees in the 46-cm irrigation treatment were only half the size of trees in the 137-cm irrigation treatment. These treatment differences were maintained through the fifth growing season. Although stem volumes of Clone TD trees in the 76- and 137-cm irrigation treatments were larger than the corresponding values for Clone DN trees at the end of the third growing season (1994), these clonal differences gradually decreased in subsequent years and were not detectable after 5 years, because stem volume relative growth rate of Clone DN was greater than that of Clone TD in all treatments. Although both clones exhibited similar predawn leaf water potentials, Clone DN typically maintained higher midday leaf water potentials, suggesting better stomatal control of water loss. Clonal and treatment differences in osmotic potential at full turgor were minimal and could not explain the clonal differences in drought resistance. Root density and root density to stem volume ratio increased more in response to moderate drought in Clone DN than in Clone TD, resulting in enhanced drought resistance (high stem volume growth rate under moderate drought conditions) and an increased capacity to withdraw water from the soil. We conclude that the greater drought resistance of Clone DN compared with Clone TD was the result of the maintenance of a more favorable water balance by stomatal regulation and greater carbon allocation to roots during the early stages of drought. However, the low root density to stem volume ratio in Clone DN growing in the 46-cm irrigation treatment suggests that severe water limitation restricted the preferential allocation of carbon to belowground tissues, so that both root and shoot growth were constrained by severe drought. PMID- 12651415 TI - Metabolism of deuterium- and tritium-labeled gibberellins in cambial region tissues of Eucalyptus globulus stems. AB - Deuterium- and tritium-labeled gibberellins (GAs) were applied to stems of 3-year old Eucalyptus globulus Labill. saplings and 9-month-old potted seedlings. Cambial region tissues surrounding the application point were collected 6, 24 or 48 h later. Twenty-four hours after application of 5 &mgr;g of [(2)H(2)]GA(20), 7% of the cambial region GA(20) pool, 7% of the GA(1) pool and 58% of the GA(29) pool were labeled with deuterium based on selected ion monitoring of purified extracts subjected to gas chromatography-mass spectrometry. The relatively low percent dilution of endogenous GAs by [(2)H(2)]GAs suggests that the exogenous application of [(2)H(2)]GA(20) did not result in substrate overloading, indicating that these conversions probably occur naturally within cambial region tissues. Extracts from similar cambial region tissues fed tritium-labeled GAs were sequentially fractionated by SiO(2) partition chromatography, C(18) reversed phase HPLC and N(CH(3))(2) HPLC. The radioactivity profiles indicated metabolism of GA(20) to GA(1) and GA(29), GA(1) conversion to GA(8), GA(4) to GA(34) and GA(9) to GA(51). Gibberellins GA(34), GA(51) and GA(29) are C-2beta-hydroxylated catabolites of low biological activity, whereas GA(1) and GA(4) are probably effectors of growth in the Eucalyptus stem and shoot. Evidence for C-13 hydroxylation of GA(4) to GA(1), GA(9) to GA(4) or GA(9) to GA(20) in the stem was inconclusive. Thus, although GA(4) and GA(9) are native to cambial region tissues, GA(1) is probably not produced from them in significant quantities. We conclude that the early C-13-hydroxylation pathway; i.e., conversion of GA(19) to GA(20) to GA(1), is the major pathway of GA(1) biosynthesis. PMID- 12651416 TI - Variation in sugar maple root respiration with root diameter and soil depth. AB - Root respiration may account for as much as 60% of total soil respiration. Therefore, factors that regulate the metabolic activity of roots and associated microbes are an important component of terrestrial carbon budgets. Root systems are often sampled by diameter and depth classes to enable researchers to process samples in a systematic and timely fashion. We recently discovered that small, lateral roots at the distal end of the root system have much greater tissue N concentrations than larger roots, and this led to the hypothesis that the smallest roots have significantly higher rates of respiration than larger roots. This study was designed to determine if root respiration is related to root diameter or the location of roots in the soil profile. We examined relationships among root respiration rates and N concentration in four diameter classes from three soil depths in two sugar maple (Acer saccharum Marsh.) forests in Michigan. Root respiration declined as root diameter increased and was lower at deeper soil depths than at the soil surface. Surface roots (0-10 cm depth) respired at rates up to 40% greater than deeper roots, and respiration rates for roots < 0.5 mm in diameter were 2.4 to 3.4 times higher than those for roots in larger diameter classes. Root N concentration explained 70% of the observed variation in respiration across sites and size and depth classes. Differences in respiration among root diameter classes and soil depths appeared to be consistent with hypothesized effects of variation in root function on metabolic activity. Among roots, very fine roots in zones of high nutrient availability had the highest respiration rates. Large roots and roots from depths of low nutrient availability had low respiration rates consistent with structural and transport functions rather than with active nutrient uptake and assimilation. These results suggest that broadly defined root classes, e.g., fine roots are equivalent to all roots < 2.0 mm in diameter, do not accurately reflect the functional categories typically associated with fine roots. Tissue N concentration or N content (mass x concentration N) may be a better indicator of root function than root diameter. PMID- 12651417 TI - Effects of elevated CO(2) and temperature on cold hardiness and spring bud burst and growth in Douglas-fir (Pseudotsuga menziesii). AB - We examined effects of elevated CO(2) and temperature on cold hardiness and bud burst of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings. Two-year old seedlings were grown for 2.5 years in semi-closed, sunlit chambers at either ambient or elevated (ambient + ~ 4 degrees C) air temperature in the presence of an ambient or elevated (ambient + ~ 200 ppm) CO(2) concentration. The elevated temperature treatment delayed needle cold hardening in the autumn and slowed dehardening in the spring. At maximum hardiness, trees in the elevated temperature treatment were less hardy by about 7 degrees C than trees in the ambient temperature treatment. In general, trees exposed to elevated CO(2) were slightly less hardy during hardening and dehardening than trees exposed to ambient CO(2). For trees in the elevated temperature treatments, date to 30% burst of branch terminal buds was advanced by about 6 and 15 days in the presence of elevated CO(2) and ambient CO(2), respectively. After bud burst started, however, the rate of increase in % bud burst was slower in the elevated temperature treatments than in the ambient temperature treatments. Time of bud burst was more synchronous and bud burst was completed within a shorter period in trees at ambient temperature (with and without elevated CO(2)) than in trees at elevated temperature. Exposure to elevated temperature reduced final % bud burst of both leader and branch terminal buds and reduced growth of the leader shoot. We conclude that climatic warming will influence the physiological processes of dormancy and cold hardiness development in Douglas-fir growing in the relatively mild temperate region of western Oregon, reducing bud burst and shoot growth. PMID- 12651418 TI - An analysis of light effects on foliar morphology, physiology, and light interception in temperate deciduous woody species of contrasting shade tolerance. AB - Maximum Rubisco activities (V(cmax)), rates of photosynthetic electron transport (J(max)), and leaf nitrogen and chlorophyll concentrations were studied along a light gradient in the canopies of four temperate deciduous species differing in shade tolerance according to the ranking: Populus tremula L. < Fraxinus excelsior L. < Tilia cordata Mill. = Corylus avellana L. Long-term light environment at the canopy sampling locations was characterized by the fractional penetration of irradiance in the photosynthetically active spectral region (I(sum)). We used a process-based model to distinguish among photosynthesis limitations resulting from variability in fractional nitrogen investments in Rubisco (P(R)), bioenergetics (P(B), N in rate-limiting proteins of photosynthetic electron transport) and light harvesting machinery (P(L), N in chlorophyll and thylakoid chlorophyll-protein complexes). On an area basis, V(cmax) and J(max) (V(a) (cmax) and J(a) (max)) increased with increasing growth irradiance in all species, and the span of variation within species ranged from two (T. cordata) to ten times (C. avellana). Examination of mass-based V(cmax) and J(max) (V(m) (cmax) and J(m) (max)) demonstrated that the positive relationships between area-based quantities and relative irradiance mostly resulted from the scaling of leaf dry mass per area (M(A)) with irradiance. Although V(m) (cmax) and J(m) (max) were positively related to growth irradiance in C. avellana, and J(m) (max) was positively related to irradiance in P. tremula, the variation range was only a factor of two. Moreover, V(m) (cmax) and J(m) (max) were negatively correlated with relative irradiance in T. cordata. Rubisco activity in crude leaf extracts generally paralleled the gas-exchange data, but it was independent of light in T. cordata, suggesting that declining V(m) (cmax) with increasing relative irradiance was related to increasing diffusive resistances from the intercellular air spaces to the sites of carboxylation in this species. Because irradiance had little effect on foliar nitrogen concentration, the relationships of P(B) and P(R) with irradiance were similar to those of V(m) (cmax) and J(m) (max). Shade intolerant species tended to have greater P(B) and P(R) and also larger V(a) (cmax) and J(a) (max) than more shade-tolerant species. However, for the whole material, P(B) and P(R) varied only about 50%, whereas V(a) (cmax) and J(a) (max) varied more than 15-fold, further emphasizing the importance of leaf anatomical plasticity in determining photosynthetic acclimation to high irradiance. Leaf chlorophyll concentrations and fractional nitrogen investments in light harvesting increased hyperbolically with decreasing irradiance to improve quantum use efficiency for incident irradiance. The effect of irradiance on P(L) was of the same order as its effect in the opposite direction on M(A), leading to either a constant model estimate of leaf absorptance with I(sum) or a slightly positive correlation. We conclude that leaf morphological plasticity is a more relevant determinant of foliage adaptation to high irradiance than foliage biochemical properties, whereas biochemical adaptation to low irradiance is of the same magnitude as the anatomical adjustments. Although shade-tolerant species did not have greater chlorophyll concentrations and P(L) than shade-intolerant species, they possessed lower M(A), and could maintain a more extensive foliar display for light capture with constant biomass investment in leaves. PMID- 12651419 TI - A comparison of heat pulse and deuterium tracing techniques for estimating sap flow in Eucalyptus grandis trees. AB - Sap flow rates were measured simultaneously by the heat pulse and deuterium tracing techniques in nine Eucalyptus grandis W. Hill ex Maiden. trees at two sites (1) to compare results from the two techniques and (2) to assess the impact of the assumptions underlying the deuterium tracing method on the calculation of sap flow for a range of tree sizes. The trees ranged in height from 4 to 14 m with leaf areas of 5 to 35 m(2). In all trees, sap flow estimated by the deuterium tracing technique was higher than sap flow estimated by the heat pulse method, with differences of 11 to 43% in eight of the trees and 113% in one tree. The largest difference was attributed to errors in the heat pulse method, as indicated by aberrant relationships between sap flow measured by the heat pulse method and tree size characteristics (i.e., diameter, sap wood area, leaf area) for that tree compared with the other experimental trees. Drilling holes in the trees to allow injection of deuterium had no significant effect on sap flow, even when 32 holes were drilled. Sap flow measured by the heat pulse method was only lower after drilling than before drilling in three trees, and the difference only persisted for about 1 h. Deuterium concentrations of water collected from the tree canopies had not returned to background values 17 days after injection. Twenty-one days after injection, sapwood and heartwood samples taken from trunks near the injection sites contained considerable concentrations of deuterium, indicating that some of the deuterium injected into the trees was still present. An experiment performed on two trees showed that deuterium was stored in the heartwood and sapwood throughout the trees, and its distribution within the trees four days after injection was similar whether it was injected into only the sapwood (where it should mix with sap and be transported from the tree most readily) or into both the sapwood and heartwood, indicating that there was considerable movement of deuterium between the heartwood and sapwood. Deuterium storage was accounted for by an approximate means in the sap flow calculations, and may have resulted in an error of about 10% in sap flow estimated by this method. We conclude that the heat pulse and deuterium tracing techniques can be used simultaneously to increase the number of sap flow measurements obtained from a forest, thereby increasing the precision of forest water use estimates. Their combination would be most effective in stands with a wide range of tree sizes and sap flow rates, where the relative differences in sap flux estimates between the methods is small compared with differences in sap flow between trees. PMID- 12651420 TI - Effects of dormancy and environmental factors on timing of bud burst in Betula pendula. AB - We tested three theories predicting the timing of bud burst in mature birch (Betula pendula Roth) trees utilizing a 60-year phenological time series together with meteorological temperature observations. Predictions of the timing of bud burst based on light conditions in addition to temperature were more accurate than predictions based on dormancy development and temperature (prediction standard error of 2.4 days versus 4.3 days). The signal from light conditions, represented by fixed calendar date, determined the start of bud ontogenesis rather than dormancy release. We suggest that models developed to predict the timing of bud burst be utilized in the analysis of plant responses to climate change and of climate change itself. PMID- 12651421 TI - Spatial and seasonal variability of photosynthetic parameters and their relationship to leaf nitrogen in a deciduous forest. AB - We used gas exchange techniques to estimate maximum rate of carboxylation (V(cmax)), a measure of photosynthetic capacity, in the understory and upper crown of a closed deciduous forest over two seasons. There was extensive variability in photosynthetic capacity as a result of vertical canopy position, species type, leaf age and drought. Photosynthetic capacity was greater in oaks than in maples and greater in the overstory than in the understory. Parameter V(cmax) was maximal early in the season but declined slowly throughout most of the summer, and then more rapidly during senescence. There was also an apparent decline during drought in some trees. Variability in V(cmax) as a result of species or vertical canopy gradients was described well by changes in leaf nitrogen per unit area (N(a)). However, temporal changes in V(cmax) were often poorly correlated with leaf nitrogen, especially in spring and summer and during drought. This poor correlation may be the result of a seasonally dependent fractional allocation of leaf nitrogen to Rubisco; however, we could not discount Rubisco inactivation, patchy stomatal closure or changes in mesophyll resistance. Consequently, when a single annual regression equation of V(cmax) versus N(a) was used for this site, there were substantial errors in the temporal patterns in V(cmax) that will inevitably result in modeling errors. PMID- 12651422 TI - Analyses of assumptions and errors in the calculation of stomatal conductance from sap flux measurements. AB - We analyzed assumptions and measurement errors in estimating canopy transpiration (E(L)) from sap flux (J(S)) measured with Granier-type sensors, and in calculating canopy stomatal conductance (G(S)) from E(L) and vapor pressure deficit (D). The study was performed in 12-year-old Pinus taeda L. stands with a wide range in leaf area index (L) and growth rate. No systematic differences in J(S) were found between the north and south sides of trees. However, J(S) in xylem between 20 and 40 mm from the cambium was 50 and 39% of J(S) in the outer 20-mm band of xylem in slow- and fast-growing trees, respectively. Sap flux measured in stems did not lag J(S) measured in branches, and time and frequency domain analyses of time series indicated that variability in J(S) in stems and branches is mostly explained by variation in D. Therefore, J(S) was used to estimate transpiration, after accounting for radial patterns. There was no difference between D and leaf-to-air vapor pressure gradient, and D did not have a vertical profile in stands of either low or high L suggesting a strong canopy atmosphere coupling. Therefore, D estimated at one point in the canopy can be used to calculate G(S) in such stands. Given the uncertainties in J(S), relative humidity, and temperature measurements, to keep errors in G(S) estimates to less than 10%, estimates of G(S) should be limited to conditions in which D >/= 0.6 kPa. PMID- 12651423 TI - Temporal and spatial variation in cyanogenic glycosides in Eucalyptus cladocalyx. AB - The release of hydrogen cyanide from endogenous cyanide-containing compounds in plants is an effective herbivore deterrent. We investigated temporal and spatial variations in cyanogenic glycoside concentration in greenhouse-grown seedlings and 6-year-old plantation trees of Eucalyptus cladocalyx F. Muell., which allocates up to 20% of leaf nitrogen to the cyanogenic glycoside, prunasin. The highest cyanogenic glycoside concentrations were in the young, developing vegetative and reproductive tissues. Both the overall cyanogenic glycoside concentration and the proportion of nitrogen allocated to cyanogenic glycoside decreased as tissues matured. Cyanogenic glycoside and nitrogen concentrations were similar at all positions on the leaf blade. There was no change in concentration of cyanogenic glycosides either diurnally or following wounding of the tissue, suggesting that these compounds are constitutive. Cyanogenic glycoside concentration varied seasonally in young leaf tips of field-grown E. cladocalyx, but not in mature, fully expanded leaves. Although some of the changes in cyanogenic glycoside concentration in young leaf tips may have been driven by changes in leaf nitrogen, there was a significant decrease in the proportion of nitrogen allocated to cyanogenic glycosides in young leaves during the summer, coinciding with the peak flowering period. Mobilization of cyanogenic glycosides may have occurred to provide nitrogen for reproduction. Most of the observed temporal and spatial variations in cyanogenic glycosides are consistent with the optimal use of resources, particularly nitrogen. PMID- 12651424 TI - Effects of continuous optimal fertilization on belowground ectomycorrhizal community structure in a Norway spruce forest. AB - Studies of effects of fertilizer treatment on ectomycorrhizal fungal community structure have predominantly been based on large, single additions of nitrogen. Studies involving chronic additions of nutrients in combination with irrigation are much less common. We used morphotyping to study effects of balanced additions of a nutrient solution on ectomycorrhizal fungal community structure in a 36-year old stand of Picea abies (L.) Karst. Despite high variability among individual samples, principal components analysis revealed a clear shift in community structure in response to fertilization. Irrigated plots receiving only water did not differ significantly from untreated control plots. Mycorrhizal root tips colonized by Cenococcum geophilum Fr. were significantly more common in fertilized plots than in control plots. Possible responses by other ectomycorrhizal species were masked by high variability. Over sixty morphotypes were distinguished, but there was no measurable effect of either fertilizer or irrigation treatment on morphotype richness or total number of root tips. PMID- 12651425 TI - Genetic transformation of silver birch (Betula pendula) by particle bombardment. AB - We used in vitro callus and shoot cultures as target material for genetic transformation of silver birch (Betula pendula Roth) by particle bombardment. Cultivation of in vitro shoot cultures before particle bombardment and a long selection period, combined with a high concentration of selective agent after bombardment, led to the production of transformed plantlets that were stable, and no escapes were found among the tree lines produced. Clonal variation in transformation efficiency was found in transient expression of the beta glucuronidase gene in callus cultures and in plantlets transformed by stable integration of the ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) and neomycin phosphotransferase (npt2) genes. PMID- 12651426 TI - Influence of temperature on the dynamics of ATP, ADP and non-adenylic triphosphate nucleotides in vegetative and floral peach buds during dormancy. AB - The nucleotides test of endodormancy, which is based on the capacity of tissues to synthesize ATP and non-adenylic triphosphate nucleotides (NTP), cannot be used for floral buds, and it is of questionable use for vegetative buds. In an attempt to find an alternative test, we examined whether the dormancy state of vegetative and floral buds of trees exposed to different temperature conditions during the rest period is directly related to their ATP, ADP and NTP concentrations and ATP/ADP ratio. Once the buds had entered endo- or paradormancy, the nucleotide concentrations and the ATP/ADP ratio were low in the vegetative primordia and very low in the floral primordia. Only after the action of chilling, when the buds were considered to have completed the endodormancy and paradormancy phases, did the nucleotide concentrations increase, accompanied by a steep rise in ATP/ADP ratio. We conclude that the ATP/ADP ratio could be used to characterize the bud dormancy state by comparison with critical values of 1.5 for vegetative primordia and 1.0 for floral primordia. PMID- 12651427 TI - Seasonal alteration in amount of Ca(2+) in apical bud cells of mulberry (Morus bombciz Koidz): an electron microscopy-cytochemical study. AB - Subcellular localization of calcium in apical bud cells of mulberry (Morus bombciz Koidz) was investigated by a combination of calcium antimonate precipitation and electron microscopy (EM), over a 10-month period extending from July to May. Calcium antimonate deposits, an indication of Ca(2+) localization, were found mainly in the intercellular spaces and vacuoles of tissues collected in summer (e.g., July 10). Few deposits were seen in the cytosol and the nucleus, indicating that when plants were actively growing, the cytoplasm had a low concentration of Ca(2+). As the day length became shorter (e.g., August 8), Ca(2+) deposits increased in the cytosol and the nucleus. When plant dormancy and cold hardiness were rapidly developing in September and October, a large number of Ca(2+) deposits were seen in the cytoplasm and the nucleus, where they remained for about 60 days. Tissue samples collected during midwinter (e.g., January 20) displayed few Ca(2+) deposits in the cytoplasm and the nucleus compared with the sample collected on November 1. The number of Ca(2+) deposits remained low even when day length increased in the spring (e.g., May 5). We conclude that the seasonal dynamics of intracellular Ca(2+) concentration bear a close relationship with growth cessation and the development of dormancy and cold hardiness in temperate woody perennials. PMID- 12651428 TI - Variation in leaf trichomes of Wigandia urens: environmental factors and physiological consequences. AB - Seasonal and environmentally induced variation in the type and frequency of leaf trichomes of Wigandia urens (Ruiz & Pavon) Kunth (Hydrophyllaceae) was studied. Depending on the microsite, W. urens plants had smooth leaves with glandular trichomes or bristly leaves with both glandular trichomes and urticant trichomes (stinging hairs). Trichome density (number of urticant trichomes per unit leaf area) was higher in the dry season than in the wet season, and was significantly correlated with both temperature (r = 0.353, P < 0.05) and photosynthetic active radiation (r = 0.313, P < 0.05). Plants established in sun-exposed areas had trichome densities three times higher than those of plants established in shaded areas during the dry season, and 28 times higher during the wet season. At both exposed and shaded sites, trichome densities of the youngest leaves of young plants were higher than those of the youngest leaves of mature plants. In smooth and bristly leaves, transpiration rates decreased with increasing temperature during the day. However, smooth leaves had higher transpiration rates than bristly leaves at both exposed and shaded sites. In laboratory studies, trichome density was significantly (P < 0.01) reduced when small sun-grown plants (0.20 0.30 m tall) were either shaded or irrigated. In larger plants, also, irrigation significantly (P < 0.01) reduced trichome density relative to that of unirrigated controls. PMID- 12651429 TI - Detection of a chitinase-like protein in the roots of Douglas-fir trees infected with Armillaria ostoyae and Phellinus weirii. AB - Protein was extracted from root bark of 11- and 25-year-old interior Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) trees that were naturally infected with Armillaria ostoyae (Romagnesi) Herink. The proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Root bark tissue adjacent to infected areas had a significantly higher protein concentration than healthy tissue (P < 0.05), whereas the protein concentration of infected tissue was consistently lower (P < 0.05) than that of healthy tissue. The SDS-PAGE profiles of healthy, infected, and adjacent-to-infected root bark tissues revealed significant differences in concentrations of a 29.3-kDa protein. The N terminal amino acid sequence of the 29.3-kDa protein displayed significant homology (P = 0.013) to a basic endochitinase. Use of a polyclonal antibody raised against the 29.3-kDa putative endochitinase-like protein (ECP) indicated differences in the quantities of ECP in healthy roots compared with roots infected with A. ostoyae in 11- and 25-year-old interior Douglas-fir trees. The antibody was also used to screen for the presence of the 29.3-kDa protein in roots of 24-year-old coastal Douglas-fir (Pseudotsuga menziesii var. menziesii) trees that were artificially inoculated with and colonized by Phellinus weirii (Murr.) Gilbn. The amount of ECP was elevated in root bark of coastal Douglas-fir in response to P. weirii infection, although in lower quantities relative to those found in the A. ostoyae-interior Douglas-fir pathosystem. The sequence homology of the ECP with a basic chitinase, together with its increased synthesis in response to two fungal pathogens, indicate a possible role for this protein in the defense of Douglas-fir against fungal pathogens. PMID- 12651430 TI - Genes expressed in the latex of Hevea brasiliensis. AB - Rubber (cis-1,4-polyisoprene), an important raw material for many industrial uses, is synthesized in the latex of Hevea brasiliensis (Willd. ex A. Juss.) Mull. Arg. We postulated that the genes uniquely or preferentially expressed in the latex may be important for rubber biosynthesis. We constructed cDNA libraries from the latex of H. brasiliensis to investigate the genes expressed in the latex by single-run partial sequencing of the cDNA clones. Sequence analyses identified 245 expressed sequence tags (ESTs), of which 57% showed homology to previously described sequences in public databases. About 16% of the database-matched ESTs encode rubber biosynthesis-related proteins such as rubber elongation factor (REF) and small rubber particle protein (SRPP). The second most frequent transcripts next to rubber biosynthesis-related genes were defense genes and protein metabolism-related genes (12.6% each). About 27% of the database-matched ESTs had sequence homology with genes of unknown function. Among the redundantly expressed genes, REF was the most frequently expressed (6.1%), followed by SRPP (3.7%) and HbLAR (2.9%). Northern blot analyses showed that ten (71%) of the 14 ESTs studied were expressed at a higher level in latex than in leaves. PMID- 12651431 TI - Radial variation in sap velocity as a function of stem diameter and sapwood thickness in yellow-poplar trees. AB - Canopy transpiration and forest water use are frequently estimated as the product of sap velocity and cross-sectional sapwood area. Few studies, however, have considered whether radial variation in sap velocity and the proportion of sapwood active in water transport are significant sources of uncertainty in the extrapolation process. Therefore, radial profiles of sap velocity were examined as a function of stem diameter and sapwood thickness for yellow-poplar (Liriodendron tulipifera L.) trees growing on two adjacent watersheds in eastern Tennessee. The compensation heat pulse velocity technique was used to quantify sap velocity at four equal-area depths in 20 trees that ranged in stem diameter from 15 to 69 cm, and in sapwood thickness from 2.1 to 14.8 cm. Sap velocity was highly dependent on the depth of probe insertion into the sapwood. Rates of sap velocity were greatest for probes located in the two outer sapwood annuli (P1 and P2) and lowest for probes in closest proximity to the heartwood (P3 and P4). Relative sap velocities averaged 0.98 at P1, 0.66 at P2, 0.41 at P3 and 0.35 at P4. Tree-specific sap velocities measured at each of the four probe positions, divided by the maximum sap velocity measured (usually at P1 or P2), indicated that the fraction of sapwood functional in water transport (f(S)) varied between 0.49 and 0.96. There was no relationship between f(S) and sapwood thickness, or between f(S) and stem diameter. The fraction of functional sapwood averaged 0.66 +/- 0.13 for trees on which radial profiles were determined. No significant depth related differences were observed for sapwood density, which averaged 469 kg m( 3) across all four probe positions. There was, however, a significant decline in sapwood water content between the two outer probe positions (1.04 versus 0.89 kg kg(-1)). This difference was not sufficient to account for the observed radial variation in sap velocity. A Monte-Carlo analysis indicated that the standard error in estimated mean f(S) declined rapidly with increasing sample size. At n = 10, the coefficient of variation in mean f(S) was 7% and at n = 15 it was slightly less than 5%. These observations indicate that radial variation in sap velocity is an important, albeit often overlooked, source of uncertainty in the scaling process. Failure to recognize that not all sapwood is functional in water transport will introduce systematic bias into estimates of both tree and stand water use. Future studies should devise sampling strategies for assessing radial variation in sap velocity and such strategies should be used to identify the magnitude of this variation in a range of non-, diffuse- and ring-porous trees. PMID- 12651432 TI - Leaf optical properties in Venezuelan cloud forest trees. AB - Leaf optical properties and related leaf characteristics were compared for thirteen cloud forest tree species differing in successional status. Sun leaves were sampled for the eight pioneer species and sun and shade leaves were sampled for the five climax species. Sun leaves had a slightly higher absorptance than shade leaves, although differences were small. Sun leaves had a higher leaf mass per unit area (LMA) and a lower chlorophyll concentration per unit leaf mass, resulting in similar chlorophyll concentrations per unit leaf area and hence similar light harvesting capacities as shade leaves. However, shade leaves realized a higher efficiency of absorptance per unit leaf biomass than sun leaves. There were few differences in leaf characteristics of sun leaves between the climax and pioneer species. Absorptance values of cloud forest species were comparable with values reported for rain forest and more seasonal forest species. Intraspecific variation in leaf absorptance was largely the result of variation in LMA, whereas interspecific variation in leaf absorptance was largely a result of variation in chlorophyll concentration per unit leaf area. PMID- 12651433 TI - Needle and stem wood production in Scots pine (Pinus sylvestris) trees of different age, size and competitive status. AB - We studied effects of tree age, size and competitive status on foliage and stem production of 43 Scots pine (Pinus sylvestris L.) trees in southern Finland. The tree attributes related to competition included foliage density, crown ratio and height/diameter ratio. Needle mass was considered to be the primary cause of growth through photosynthesis. Both stem growth and foliage growth were strongly correlated with foliage mass. Consequently, differences in growth allocation between needles and stem wood in trees of different age, size, or position were small. However, increasing relative height increased the sum of stem growth and foliage growth per unit foliage mass, indicating an effect of available light. Suppressed trees seemed to allocate more growth to stem wood than dominant trees, and their stem growth per unit foliage mass was larger. Similarly, trees in dense stands allocated more growth to stem wood than trees in sparse stands. The results conformed to the pipe model theory but seemed to contradict the priority principle of allocation. PMID- 12651434 TI - Thigmomorphogenesis: changes in the morphology and mechanical properties of two Populus hybrids in response to mechanical perturbation. AB - To identify hybrid-specific differences in developmental response to mechanical perturbation (MP), we compared the effects of stem flexure on several morphological and mechanical properties of two Populus trichocarpa Torr. & A. Gray x P. deltoides Bartr. ex Marsh. hybrids, 47-174 and 11-11. In response to the MP treatment, both hybrids exhibited a significant increase in radial growth, especially in the direction of the MP (47-174, P = 0.0001; 11-11, P = 0.002), and a significant decrease in height to diameter growth ratio (P = 0.0001 for both hybrids), suggesting that MP-treated stems are more tapered than control stems. A direct consequence of the MP-induced increase in radial growth was a significant increase in flexural rigidity (EI, N mm(2)) in stems of both hybrids (47-174, P = 0.0001; 11-11, P = 0.009). Both control and MP-treated stems of Hybrid 47-174 had significantly greater height to diameter ratios and EI values than the corresponding stems of Hybrid 11-11 (11-11 stem ratios and EI values were 85 and 76%, respectively, of those of 47-174). In Hybrid 47-174, Young's modulus of elasticity (E, N mm(-2)), a measure of stem flexibility, for MP-treated stems was only 80% of the control value (P = 0.0034), whereas MP had no significant effect on E of stems of Hybrid 11-11 (P = 0.2720). Differences in flexure response between the hybrids suggest that Hybrid 47-174 can produce a stem that is more tolerant of wind-induced flexure by altering both stem allometry and material properties, whereas Hybrid 11-11 relies solely on changes in stem allometry for enhanced stability under MP conditions. PMID- 12651435 TI - Effects of xylem cavitation and freezing injury on dieback of yellow birch (Betula alleghaniensis) in relation to a simulated winter thaw. AB - Shoot dieback, shoot growth, stem xylem cavitation, stem and root freezing injury, and root pressure were measured in 2-year-old, cold-hardened, potted yellow birch (Betula alleghaniensis Britt.) seedlings that had been subjected to a simulated winter thaw for 0, 5, 10, 19 or 27 days followed by 10 weeks at -10 degrees C. Stem xylem cavitation was determined as percent loss of hydraulic conductivity. Stem freezing injury was measured as electrolyte leakage (EL). Root freezing injury was determined by EL and by triphenyl tetrazolium chloride (TTC) reduction. Thaw duration was significantly correlated with dieback, new shoot growth, stem xylem cavitation, stem and root freezing damage, and root pressure (P < 0.05). In particular, shoot dieback was positively correlated with stem xylem cavitation (P < 0.001), residual stem xylem cavitation (P < 0.01) and root freezing injury (P < 0.010), but only weakly correlated with stem freezing damage (P < 0.05). In roots, freezing damage was negatively correlated with root pressure (P < 0.05), which, in turn, was negatively correlated with residual stem xylem cavitation after root pressure development. In stems, there was no correlation between freezing damage and xylem cavitation. We conclude that long periods of winter thaw followed by freezing resulted in freezing injury to roots concomitant with a reduction in root pressures, leading to poor recovery from freezing-induced xylem embolism. PMID- 12651436 TI - Seasonal patterns of carbohydrate reserves in red spruce seedlings. AB - We studied seasonal dynamics of carbohydrate storage in red spruce (Picea rubens Sarg.) seedlings by measuring starch and sugar concentrations of old needles (>/= one year old), new needles (< one year old), stems, and roots in two stands in the Green Mountains of Vermont. Although the two stands differed in many site characteristics including percent slope, aspect, soil type, drainage, and 564 m in elevation, concentrations and seasonal patterns of carbohydrates were similar for the two stands. For all tissues, starch concentrations peaked in late spring, declined through summer, and reached a minimum in winter. Sugar concentrations were greater than starch concentrations in all months except May and June. Sugar concentrations peaked in winter, and old needles showed a significant increase in sugar concentration between February and March. This increase in sugar concentration occurred without any reduction in localized starch concentrations or reductions in sugar or starch concentrations in new needles, stems or roots. Because March measurements were made toward the end of a prolonged thaw, a time when increases in photosynthesis have been documented for red spruce, it is likely that the March increase in sugar concentrations resulted from photosynthesis during the thaw. Compared with stems and roots, needles generally contained the highest concentration of carbohydrates and exhibited the greatest seasonal change in carbohydrate concentration. Needles were also the largest reservoir of carbohydrates throughout the year, especially during winter. Because of the critical roles of needles in photosynthesis and storage of carbohydrates, we conclude that any factors that disrupt the accumulation or availability of carbohydrates in red spruce needles will greatly alter plant carbon relations. PMID- 12651437 TI - Drought tolerance of clonal Malus determined from measurements of stomatal conductance and leaf water potential. AB - We examined tolerance to soil drying in clonally propagated apple (Malus domestica Borkh.) rootstocks used to control shoot growth of grafted scions. We measured leaf conductance to water vapor (g(L)) and leaf water potential (Psi(L)) in a range of potted, greenhouse-grown rootstocks (M9, M26, M27, MM111, AR69-7, AR295-6, AR360-19, AR486-1 and AR628-2) as the water supply was gradually reduced. Irrespective of the amount of available water, rootstocks that promoted scion shoot growth (M26 and MM111) generally had higher g(L) and more negative Psi(L) than rootstocks that restricted scion shoot growth (M27 and M9). After about 37 days of reduced water supply, there were significant decreases in g(L) and Psi(L) in all rootstocks compared with well-watered controls. In all treatments, the slope of the relationship between log (g(L)) and Psi(L) was positive, except for rootstocks AR295-6, AR628-2 and AR486-1 in the severe drought treatment, where the drought-induced change in the relationship suggests that rapid stomatal closure occurred when leaf water potentials fell below -2.0 MPa. This drought response was associated with increased root biomass production. Rootstock M26 showed little stomatal closure even when its water potential fell below -2.0 MPa, and there was no effect of drought on root biomass production. We conclude that differences among rootstocks in the way that g(L) and Psi(L) respond to drought reflect differences in the mechanisms whereby they tolerate soil drying. We suggest that these differences are related to differences among the rootstocks in their ability to control shoot growth. PMID- 12651438 TI - Effect of elevated [CO(2)] and varying nutrient application rates on physiology and biomass accumulation of Sitka spruce (Picea sitchensis). AB - Sitka spruce (Picea sitchensis (Bong.) Carr.) seedlings were supplied with solutions containing nitrogen (N) at 0.1 x or 2 x the optimum rate (low-N and high-N supply, respectively) and grown either outside in a control plot or inside open-top chambers and exposed to ambient (355 &mgr;mol mol(-1)) or elevated (700 &mgr;mol mol(-1)) CO(2) concentration ([CO(2)]). Gas exchange measurements, chlorophyll determinations and nutrient analysis were made on current-year (< 1 year-old) shoots of the upper whorl after the seedlings had been growing in the [CO(2)] treatments for 17 months and the nutrient treatments for 6 months. Total seedling biomass and biomass allocation were assessed at the end of the experiment. Nutrient treatment had a significant effect on the light response curves, irrespective of [CO(2)] or chamber treatment; seedlings supplied with high-N rates had higher net photosynthetic rates than seedlings supplied with low N rates. The degree of photosynthetic stimulation in response to elevated [CO(2)] was larger in seedlings receiving high-N rates than in seedlings receiving low-N rates. Light-saturated net photosynthesis of seedlings grown and measured in elevated [CO(2)] was 26% higher than that of seedlings grown and measured in ambient [CO(2)]. There was no significant effect of [CO(2)] or chamber treatment on the CO(2) response curves of seedlings receiving High-N supply rates. In contrast, analysis of the CO(2) response curves of seedlings receiving Low-N supply rates showed acclimation to elevated [CO(2)]. Both maximum rate of carboxylation (V(cmax)) and maximum electron transport capacity (J(max)) were lower and J(max)/V(cmax) higher in seedlings in the elevated [CO(2)] treatment. There was no effect of elevated [CO(2)] on stomatal conductance, although it was highly dependent on foliar [N], ranging from ~60 mmol m(-2) s(-1) at ~1.5 g N m( 2) to 200 mmol m(-2) s(-1) at ~5 g N m(-2). In the high-N and low-N treatments, foliar N concentration was 10 and 28% lower in seedlings grown in elevated [CO(2)] than in seedlings grown in ambient [CO(2)], respectively. There was no [CO(2)] effect on foliar phosphorus concentration ([P]). Chlorophyll concentration increased with increasing N supply in all treatments. There was no significant effect of elevated [CO(2)] on specific leaf area. Chlorophyll concentration expressed either on an area or dry mass basis for a given foliar [N] was higher in seedlings grown in elevated [CO(2)] than in seedings grown in ambient [CO(2)]. Elevated [CO(2)] increased total biomass accumulation by 37% in seedlings in the high-N treatment but had no effect in seedlings in the low-N treatment. There was a proportionally bigger allocation of biomass to roots of seedlings in the elevated [CO(2)] + low-N supply rate treatment compared with seedlings in other treatments. This resulted in a reduction in aboveground biomass compared with corresponding seedlings grown in ambient [CO(2)]. PMID- 12651439 TI - Seasonal CO(2) assimilation and stomatal limitations in a Pinus taeda canopy. AB - Net CO(2) assimilation (A(net)) of canopy leaves is the principal process governing carbon storage from the atmosphere in forests, but it has rarely been measured over multiple seasons and multiple years. I measured midday A(net) in the upper canopy of maturing loblolly pine (Pinus taeda L.) trees in the piedmont region of the southeastern USA on 146 sunny days over 36 months. Concurrent data for leaf conductance and photosynthetic CO(2) response curves (A(net)-C(i) curves) were used to estimate the relative importance of stomatal limitations to CO(2) assimilation in the field. In fully expanded current-year and 1-year-old needles, midday light-saturated A(net) was constant over much of the growing season (5-6 &mgr;mol CO(2) m(-2) s(-1)), except during drought periods. During the winter season (November-March), midday A(net) of overwintering needles varied in proportion to leaf temperature. Net CO(2) assimilation at light saturation occurred when daytime air temperatures exceeded 5-6 degrees C, as happened on more than 90% of the sunny winter days. In both age classes of foliage, winter carbon assimilation accounted for approximately 15% of the daily carbon assimilation on sunny days throughout the year, and was relatively insensitive to year-to-year differences in temperature during this season. However, strong stomatal limitations to A(net) occurred as a result of water stress associated with freezing cycles in winter. During the growing season, drought-induced water stress produced the largest year-to-year differences in seasonal CO(2) assimilation on sunny days. Seasonal A(net) was more drought sensitive in current year needles than in 1-year-old needles. Relative stomatal limitations to daily integrated A(net) were approximately 40% over the growing season, and summer drought rather than high temperatures had the largest impact on summer A(net) and integrated annual CO(2) uptake in the upper crown. Despite significant stomatal limitations, a long duration of near-peak A(net) in the upper crown, particularly in 1-year-old needles, conferred high seasonal and annual carbon gain. PMID- 12651440 TI - Vertical gradients in photosynthetic light response within an old-growth Douglas fir and western hemlock canopy. AB - We examined needle-level light response of photosynthesis across a vertical light gradient within 45-55-m-tall western hemlock (Tsuga heterophylla (Raf.) Sarg.) and Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) trees growing in a 400-500 year-old mixed species stand. We determined: (1) whether light-saturated photosynthetic rates, light compensation points, and respiration rates varied from the upper to the lower canopy, and (2) if light-saturated photosynthetic rates, light compensation points, and respiration rates varied between Douglas fir and western hemlock. Over a 25-m gradient from the canopy top to the lower canopy, mean light-saturated photosynthetic rates, light compensation points, and respiration rates declined in overstory Douglas-fir and western hemlock needles, paralleling a 65% decline in the mean daily photosynthetic photon flux density (PPFD). At the canopy top, increasing light-saturated photosynthetic rates relative to lower canopy needles increased carbon uptake at high PPFD. In the lower canopy, reduced respiration rates relative to upper canopy needles increased carbon uptake at low PPFD by reducing the light compensation point. At all canopy positions, western hemlock had lower mean light-saturated photosynthetic rates, light compensation points and respiration rates than Douglas-fir. As a result, western hemlock had higher net photosynthetic rates at low PPFD, but lower net photosynthetic rates at high PPFD compared with Douglas fir. PMID- 12651441 TI - Differential expression of genes encoding cell wall proteins in vascular tissues from vertical and bent loblolly pine trees. AB - Differential expression of cell wall proteins during plant development and in response to biotic or abiotic stress suggests that these proteins may contribute in different ways to plant cell wall architecture. Because the wood of loblolly pine (Pinus taeda L.) is highly specialized in the formation of secondary cell walls, it is an ideal tissue for studying these proteins. The cDNAs coding for six novel cell wall associated proteins, as well as a homologue for a phytocyanin, were identified and characterized from differentiating xylem of loblolly pine. Three of these cDNAs encoded new putative loblolly pine arabinogalactan proteins, based on their structural similarity to classical arabinogalactan proteins (AGPs). In addition, one clone was related to the proline-rich protein group and the other two to the glycine-rich protein group and the mussel adhesive protein. Relative expression of these genes was examined in different tissues and organs from normal trees (needles, phloem and vertical wood), the underside of bent trees (compression wood) and the lateral sides of the bent stems (side wood). All clones, except one, were highly expressed in vascular tissues with noticeable differences among the three types of wood. Their relationships and diversity provide the first insights into concerted expression and related function for this important group of proteins in cell wall formation of wood. PMID- 12651442 TI - Changes in cellular structures and enzymatic activities during browning of Scots pine callus derived from mature buds. AB - Visible browning is a typical feature of callus cultures derived from shoot tips of mature Scots pine (Pinus sylvestris L.). Because the ability of callus to regenerate is low, we determined the effect of browning on growth and changes in cellular structure during culture. Striking alterations in cellular structure were detected by LM (light microscopy), EM (electron microscopy) and SEM (scanning electron microscopy). Accumulation of phenolic substances was shown by histochemical staining. Staining for beta-glucosidase activity of soluble proteins that had been subjected to polyacrylamide gel electrophoresis indicated lignification of cells. The measured growth rate of callus was low compared with a hypothetical growth curve. Peroxidase activity increased rapidly soon after the start of the culture period, but especially between the second and third weeks of culture. At this time, the degradation of cell membranes and browning began coincident with the loss of chlorophyll. We conclude that browning is associated with cell disorganization and eventual cell death, making tissue culture of mature pine especially difficult. PMID- 12651443 TI - Managing forests for wood yield and carbon storage: a theoretical study. AB - Which forest management regimes best achieve the dual objectives of high sustained timber yield and high carbon storage, including the carbon stored in soil and wood products? A mechanistic forest ecosystem simulator, which couples carbon, nitrogen and water (Edinburgh Forest Model), was calibrated to mimic the growth of a pine plantation in a Scottish climate. The model was then run to equilibrium (1) as an undisturbed forest, (2) removing 2.5, 10, 20 or 40% of the woody biomass each year (3) removing 50% of the woody biomass every 20 years, and (4) clear-felling and replanting every 60 years as in conventional plantations in this climate. More carbon was stored in the undisturbed forest (35.2 kg C m(-2)) than in any regime in which wood was harvested. Plantation management gave moderate carbon storage (14.3 kg C m(-2)) and timber yield (15.6 m(3) ha(-1) year(-1)). Notably, annual removal of 10 or 20% of woody biomass per year gave both a high timber yield (25 m(3) ha(-1) year(-1)) and high carbon storage (20 to 24 kg C m(-2)). The efficiency of the latter regimes could be attributed (in the model) to high light interception and net primary productivity, but less evapotranspiration and summer water stress than in the undisturbed forest, high litter input to the soil giving high soil carbon and N(2) fixation, low maintenance respiration and low N leaching owing to soil mineral pool depletion. We conclude that there is no simple inverse relationship between the amount of timber harvested from a forest and the amount of carbon stored. Management regimes that maintain a continuous canopy cover and mimic, to some extent, regular natural forest disturbance are likely to achieve the best combination of high wood yield and carbon storage. PMID- 12651444 TI - Genetic diversity and bark physiology of the European beech (Fagus sylvatica): a coevolutionary relationship with the beech scale (Cryptococcus fagisuga). AB - In 1994 and 1995, the degree of infestation by the beech scale (Cryptococcus fagisuga Lind.) was recorded on 120 beech trees (Fagus sylvatica L.) in the Pless Forest near Gottingen, Germany. Simultaneously, the trees were characterized genetically and compounds of primary and secondary metabolism of beech bark were analyzed. A correlation was established between beech scale infestation and the genotype of the host trees, based on gene locus A of isocitrate dehydrogenase (IDH-A). The fraction of infested beech trees was higher in the heterozygous genotype A2A3 group than in the homozygous genotype A2A2 and A3A3 groups, whereas the fraction of beech trees with decreasing infestation from one year to the next was lower in the heterozygous genotype A2A3 group than in the homozygous genotype A2A2 and A3A3 groups. Concentrations of soluble carbohydrates, protein amino acids and proanthocyanidins were determined in the inner and outer bark of trees with differing degrees of infestation. The results indicate that the defense reaction of beech against infestation by beech scale comprises multiple processes in which nutrient availability to the beech scale is reduced by concentration shifts and by the formation of inhibiting compounds. PMID- 12651445 TI - Process-based models for forest ecosystem management: current state of the art and challenges for practical implementation. AB - Recent progress toward the application of process-based models in forestmanagement includes the development of evaluation and parameter estimation methods suitable for models with causal structure, and the accumulation of data that can be used in model evaluation. The current state of the art of process modeling is discussed in the context of forest ecosystem management. We argue that the carbon balance approach is readily applicable for projecting forest yield and productivity, and review several carbon balance models for estimating stand productivity and individual tree growth and competition. We propose that to develop operational models, it is necessary to accept that all models may have both empirical and causal components at the system level. We present examples of hybrid carbon balance models and consider issues that currently require incorporation of empirical information at the system level. We review model calibration and validation methods that take account of the hybrid character of models. The operational implementation of process-based models to practical forest management is discussed. Methods of decision-making in forest management are gradually moving toward a more general, analytical approach, and it seems likely that models that include some process-oriented components will soon be used in forestry enterprises. This development is likely to run parallel with the further development of ecophysiologically based models. PMID- 12651446 TI - Developing adaptive forest management strategies to cope with climate change. AB - Numerous investigations have indicated that projected climate change will impact strongly on forest growth and composition. To adapt managed forests to changing environmental conditions it may be necessary to modify traditional forest management strategies. An extended version of a forest gap model was applied to a managed forest district in northeastern Germany. The model was initialized with forest inventory data and run using routines devised to simulate three management scenarios: (1) maximized timber production, (2) climatically well-adapted forest composition, and (3) maximized tree species diversity. The strategies were compared with a baseline scenario of traditional management without any response to climate change. The comparisons were based on simulated wood production and species composition after 110 years of development. The results underline the important influence that management strategies have on forest growth. Forest management may adopt a variety of strategies to respond to the expected changes in climate. Process-oriented forest gap models can aid in the assessment of these strategies. PMID- 12651447 TI - Forest growth and species distribution in a changing climate. AB - Climate change has many potential effects on plants, some detrimental to growth, others beneficial. Increasing CO(2) concentration can increase photosynthetic rates, with the greatest increases likely to be in C(3) plants growing in warm dry conditions. Increasing temperature directly affects plant growth through effects on photosynthetic and respiration rates. However, plants have a considerable ability to adapt to changing conditions and can tolerate extremely high temperatures, provided that adequate water is available. Increasing temperature may increase vapor pressure deficits of the air, and thereby increase transpiration rates from most plant canopies. Effects are likely to vary among plant communities, with forests generally experiencing greater increases in transpiration rates than grasslands. These increases in transpiration are likely to be reduced by stomatal closure in response to increasing CO(2) concentration. In many areas, precipitation will probably increase with global warming; however, these increases may be insufficient to meet the increased transpirational demand by plant canopies. Increasing temperature is likely to increase soil organic matter decomposition rates so that nutrients may be more readily mineralized and made available to plants. In highly fertile systems, this could lead to nutrient losses through leaching. For different combinations of increases in temperature and CO(2) concentration, and for systems primarily affected by water or nutrient limitations, different overall effects on plant productivity can be expected. Responses will be negative in some circumstances and positive in others, but on the whole, catastrophic changes to forest growth seem unlikely under most conditions. In contrast,ecological consequences of climate change are potentially more serious. The distribution of many species tends to be limited to a narrow range of environmental conditions. Climate conditions over much of a species' current natural range may therefore become unsuitable, leading to significant decline of forests or of particular species within forests. PMID- 12651448 TI - Light absorption and bole volume growth of individual Douglas-fir trees. AB - Empirical growth and yield models for forest management are evolving toward individual-tree models that are capable of simulating the growth of mixed and uneven-aged stands. Spatially explicit (i.e., distance-dependent) models usually modify the growth of trees by means of competition indices; however, these competition indices rarely simulate the light available for tree growth explicitly. We used tree growth data from an even-aged, unthinned, 50-year-old Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) stand in British Columbia to test the hypothesis that the amount of absorbed light is a good predictor of diameter at breast height, height, and bole volume growth of an individual tree. We also explored the relationships between these variables. A spatially explicit light model was used to simulate photosynthetically active radiation absorbed by individual trees during a growth period (APAR) based on detailed canopy architecture information. For the purpose, we used a weighted leaf area (WLA) that is linearly related to APAR. Because of the integration of light absorption by a tree crown, estimates of WLA were highly correlated with leaf area for dominant trees. For suppressed trees, leaf area was a poor estimator of WLA. The relationship between WLA and bole volume growth was nonlinear, indicating a higher light-use efficiency in suppressed trees than in dominant trees. This relationship was strong enough to be useful for growth modeling. Only height growth of suppressed trees was affected by WLA. We conclude that single-tree WLA can be used as a process-oriented competition index in growth models for forest management. PMID- 12651449 TI - Modeling the influence of temperature on monthly gross primary productivity of sugar maple stands. AB - A bottom-up and a top-down model were used to estimate the effect of temperature on monthly gross primary productivity (GPP) of sugar maple (Acer saccharum Marsh.). The bottom-up model computed canopy photosynthesis at an hourly time step from detailed physiological sub-models of leaf photosynthesis and stomatal conductance. Leaf mass per area was used as a covariable to integrate photosynthesis through the canopy. The top-down model used a radiation-use efficiency coefficient to relate canopy gross photosynthesis to absorbed photosynthetically active radiation at a monthly time step. The parameters of the top-down model were estimated from simulations with the bottom-up model. Forty single-year simulations were made using records of daily maximum and minimum temperatures from weather stations selected within the natural range of sugar maple in the province of Quebec, Canada. Leaf area index was randomly varied between 4 and 10. Within a broad range of values, temperature had a minor effect on predicted monthly canopy-level GPP and its contribution to explaining the variability of GPP was low, both through its direct effect on photosynthetic processes (1.1%), and indirectly through the effect of relative humidity on stomatal conductance (4.0%). This result was unchanged when key parameters relating photosynthesis to temperature and stomatal conductance to atmospheric humidity were changed in the bottom-up model. An increase in time step from hourly to monthly resulted in a downward shift in the optimum temperature range for photosynthesis, from 30 degrees C for a leaf at saturating irradiance to 22 degrees C for the canopy at a monthly time scale. PMID- 12651450 TI - Application of volume growth and survival graphs in the evaluation of four process-based forest growth models. AB - Volume growth and survival (VGS) graphs, which show volume growth rate and risk of mortality for individual trees (or tree size classes), have been proposed as a tool for assessing the validity of models that describe the development over time of tree size distributions within forest stands. We examined the utility of the VGS method in evaluating four process-based models. The performance of the models FORSKA, 4C, MORG, and PipeQual is analyzed against long-term data from a Scots pine stand in Evo, Finland, and the models FORSKA and 4C are also assessed with respect to data from a beech stand in Fabrikschleichach, Germany. Comparison of the measurement-based VGS graphs with those produced from the model-based data shows that although the models yield similar stand-level predictions, they can differ widely in their projections of individual tree growth and size distributions. Examination of the discrepancies between models and data in the context of the VGS graphs reveals several areas in which the models could be improved. We conclude that the method is useful in model evaluation, especially if used in combination with indicators of stand structure, such as the height/diameter ratio. PMID- 12651451 TI - Volume growth and survival graphs: a method for evaluating process-based forest growth models. AB - We investigated the relationships within forest stands between tree size and (a) stem volume growth rate and (b) risk of mortality for individual trees. Values of both x and y variables were plotted relative to the largest value in the stand. We refer to the resultant presentations as relative volume growth and relative survival graphs (VGSs). A pair of VGSs can be produced readily from an individual tree growth model. It can also be constructed from consecutive sets of field measurements. Comparison of VGSs derived from model and measurement data provides a test of the validity of the components of the growth model. We have analyzed VGSs based on measurement data for Scots pine (Pinus sylvestris L.) in central Finland and for beech (Fagus sylvatica L.) in southern Germany. The graphs based on measurement data varied as a consequence of differences in competition, stand management, and tree species. We analyzed the relationship between VGSs and stand dynamics using a simple growth model. We found that different features of the VGSs imply characteristic tree size distributions in subsequent years. Thus, we conclude that if the VGSs generated by a model do not correspond to those based on field measurements, the model cannot be relied on to reproduce the development of tree-size distribution correctly. Relative growth and survival graphs thus provide a tool for evaluating complicated growth models. PMID- 12651452 TI - The process-based stand growth model Formix 3-Q applied in a GIS environment for growth and yield analysis in a tropical rain forest. AB - Managing tropical rain forests is difficult because few long-term field data on forest growth and the impact of harvesting disturbance are available. Growth models may provide a valuable tool for managers of tropical forests, particularly if applied to the extended forest areas of up to 100,000 ha that typically constitute the so-called forest management units (FMUs). We used a stand growth model in a geographic information system (GIS) environment to simulate tropical rain forest growth at the FMU level. We applied the process-based rain forest growth model Formix 3-Q to the 55,000 ha Deramakot Forest Reserve (DFR) in Sabah, Malaysia. The FMU was considered to be composed of single and independent small scale stands differing in site conditions and forest structure. Field data, which were analyzed with a GIS, comprised a terrestrial forest inventory, site and soil analyses (water, nutrients, slope), the interpretation of aerial photographs of the present vegetation and topographic maps. Different stand types were determined based on a classification of site quality (three classes), slopes (four classes), and present forest structure (four strata). The effects of site quality on tree allometry (height-diameter curve, biomass allometry, leaf area) and growth (increment size) are incorporated into Formix 3-Q. We derived allometric relations and growth factors for different site conditions from the field data. Climax forest structure at the stand level was shown to depend strongly on site conditions. Simulated successional pattern and climax structure were compared with field observations. Based on the current management plan for the DFR, harvesting scenarios were simulated for stands on different sites. The effects of harvesting guidelines on forest structure and the implications for sustainable forest management at Deramakot were analyzed. Based on the stand types and GIS analysis, we also simulated undisturbed regeneration of the logged over forest in the DFR at the FMU level. The simulations predict slow recovery rates, and regeneration times far exceeding 100 years. PMID- 12651453 TI - Application of process-based models to forest management: experience with PROMOD, a simple plantation productivity model. AB - Process-based growth models are rarely used as a basis for decisions in forest management. Reasons for this are discussed and the development and application of PROMOD, a simple process-based plantation productivity model designed to meet the needs of managers, is described. In particular,PROMOD is intended for screening prospective plantation sites on the basis of readily available input data, including site latitude, soil characteristics, and long-term climate data. Originally developed for use with Eucalyptus globulus Labill., PROMOD has since been parameterized also for Eucalyptus nitens (Deane & Maiden) Maiden and Pinus radiata D. Don. We discuss the adaptation of PROMOD to new environmental conditions and species; the sensitivity of the model's predictions to changes in model structure, the values of its physiological parameters and the accuracy and resolution of site survey information; and the use of the model both in assessing the profitability of plantation irrigation in semi-arid regions, and for generating broad-area productivity maps. PMID- 12651454 TI - Initializing a model stand for process-based projection. AB - A stand generation (or initialization) procedure is designed to furnish morphologically plausible model trees for process-based projection. The steps of the initialization are: (i) the generation of the locations of model trees and the tessellation of tree areas; (ii) the sampling of diameters from a target distribution and assignments of those diameters to model-tree locations; (iii) the calculation of the height of each model tree from its assigned diameter and the distances to its neighbors; (iv) the calculation of the crown length of each model tree from its height and distances to its neighbors; and (v) the recalculation of diameter from height and crown length. Components of dry matter are calculated from the model-tree dimensions and pipe-model theory. Process based projection with the AMORPHYS model is discussed. PMID- 12651455 TI - Effects of stand composition and thinning in mixed-species forests: a modeling approach applied to Douglas-fir and beech. AB - Models estimating growth and yield of forest stands provide important tools for forest management. Pure stands have been modeled extensively and successfully for decades; however, relatively few models for mixed-species stands have been developed. A spatially explicit, mechanistic model (COMMIX) is presented that simulates growth of mixed-species forest stands, and takes account of the effects of management on stand dynamics. Previously, it was shown that COMMIX satisfactorily reproduced the development of monospecific stands. In the present study, the model was used to simulate growth and yield in mixed stands differing in the proportions of species present. The concept of a "replacement series" was used to compare productivities of the mixed stands. The model was also used to analyze effects of thinning regimes and stand composition on productivity. Model simulations indicate that productivity of a mixed stand will generally be intermediate between the productivities of monospecific stands of the contributing species. However, stand composition, and especially thinning regime, will strongly affect stand productivity. The simulations are discussed with reference to the effects of resource partitioning, canopy stratification, complementarity, spatial pattern, crown dynamics, and phenology on the growth and yield of mixed stands. The study highlights the value of using mechanistic approaches to predict mixed stand development in relation to management regime. PMID- 12651456 TI - Comparing light interception with stand basal area for predicting tree growth. AB - Empirical and process-based tree growth models have been developedconcurrently; however, their growth predictions have rarely been compared directly. A major difference between the model types is the explicit quantification of foliage biomass as a key variable in process-based models. The aim of this work was to test if this difference has a significant impact on model behavior, especially when simulating silvicultural practices such as intensive thinning. A method was developed to evaluate leaf area and light interception of the mean tree of an even-aged stand from yield table data for Norway spruce (Picea abies (L.) Karst.) in the northern French Alps. Two scenarios were analyzed: (1) a closed stand where leaf area was limited by a maximum leaf area index-represented by young, dense stands, and (2) an open stand where leaf area was limited by the height of the crown base-represented by old, sparse stands. Light interception was calculated based on interpolation between a closed stand (Beer-Lambert law) and an isolated tree (light interception proportional to leaf area). This approach was then used to build a growth model in which competition was described by the ratio of light intercepted by a mean tree of the stand to light intercepted by an isolated tree of the same size. This process-based model was compared with a simpler empirical model in which competition was described by stand basal area. Both models fit well to yield table diameter increment data, the simpler model being slightly better. Simulation of long-term growth, interspersed with thinning, revealed differences between the models. The empirical model was sensitive to thinning and simulated a discontinuous growth pattern, whereas the model based on light calculation showed a smoother growth response to thinning. Simulations of heavy thinning in a dense stand highlighted these differences. The empirical model simulated heavy thinning in a dense stand unrealistically: after thinning, trees immediately grew as fast as trees of similar diameter in an unthinned stand at the same density. In contrast, leaf area played a regulatory role in the model based on light interception: trees with short crowns, as a result of a previous period of growth at high density, benefited little from an increase in light following thinning. It is concluded that models based on physiological or ecological processes have qualitative behaviors different from those of classical empirical models. This is especially important when models are used to make extrapolations far from reference data, for example, to forecast the long-term effect of a new silvicultural strategy. PMID- 12651457 TI - Bayesian synthesis for quantifying uncertainty in predictions from process models. AB - The Bayesian synthesis method is reviewed and judged to be useful for determining posterior distributions and interval estimates for inputs and outputs of process based forest models. The method furnishes posterior distributions of the values of a model's parameters and response variables. The method also provides estimates of correlation among the parameters and output variables. Bayesian synthesis is the only type of uncertainty analysis that affords incorporation of all the information available to the investigator, in addition to the information contained in the model itself. PMID- 12651458 TI - Estimating water use by sugar maple trees: considerations when using heat-pulse methods in trees with deep functional sapwood. AB - Accurate estimates of sapwood properties (including radial depth of functional xylem and wood water content) are critical when using the heat pulse velocity (HPV) technique to estimate tree water use. Errors in estimating the volumetric water content (V(h)) of the sapwood, especially in tree species with a large proportion of sapwood, can cause significant errors in the calculations ofsap velocity and sap flow through tree boles. Scaling to the whole-stand level greatly inflates these errors. We determined the effects of season, tree size and radial wood depth on V(h) of wood cores removed from Acer saccharum Marsh. trees throughout 3 years in upstate New York. We also determined the effects of variation in V(h) on sap velocity and sap flow calculations based on HPV data collected from sap flow gauges inserted at four depths. In addition, we compared two modifications of Hatton's weighted average technique, the zero-step and zero average methods, for determining sap velocity and sap flow at depths beyond those penetrated by the sap flow gauges. Parameter V(h) varied significantly with time of year (DOY), tree size (S), and radial wood depth (RD), and there were significant DOY x S and DOY x RD interactions. Use of a mean whole-tree V(h) value resulted in differences ranging from -6 to +47% for both sap velocity and sap flow for individual sapwood annuli compared with use of the V(h) value determined at the specific depth where a probe was placed. Whole-tree sap flow was 7% higher when calculated on the basis of the individual V(h) value compared with the mean whole-tree V(h) value. Calculated total sap flow for a tree with a DBH of 48.8 cm was 13 and 19% less using the zero-step and the zero-average velocity techniques, respectively, than the value obtained with Hatton's weighted average technique. Smaller differences among the three methods were observed for a tree with a DBH of 24.4 cm. We conclude that, for Acer saccharum: (1) mean V(h) changes significantly during the year and can range from nearly 50% during winter and early spring, to 20% during the growing season;(2) large trees have a significantly greater V(h) than small trees; (3) overall, V(h) decreases and then increases significantly with radial wood depth, suggesting that radial water movement and storage are highly dynamic; and (4) V(h) estimates can vary greatly and influence subsequent water use calculations depending on whether an average or an individual V(h) value for a wood core is used. For large diameter trees in which sapwood comprises a large fraction of total stem cross-sectional area (where sap flow gauges cannot be inserted across the entire cross-sectional area), the zero-average modification of Hatton's weighted average method reduces the potential for large errors in whole-tree and landscape water balance estimates based on the HPV method. PMID- 12651459 TI - Physiological responses of black spruce layers and planted seedlings to nutrient addition. AB - We investigated effects of nutrient addition on several physiological characteristics of 60-cm-tall black spruce (Picea mariana Mill. B.S.P.) layers (i.e., rooted branches of overstory trees) and 20-cm-tall planted seedlings on a clear-cut, N-limited boreal site. After two growing seasons, current-year and one year-old needles of fertilized trees (layers and seedlings combined) had higher net photosynthetic rates (A(n)) and maximum capacity of Rubisco for CO(2) fixation (V(max)) than unfertilized trees. One-year-old needles of fertilized trees had higher stomatal conductance (g(s)), higher water-use efficiency, and lower intercellular to ambient CO(2) ratio than unfertilized trees. Additionally, fertilized trees had higher predawn and midday shoot water potentials than unfertilized trees. Stomatal conductance of 1-year-old needles was 23% higher in seedlings than in layers, but there were no significant differences in g(s) of current-year needles between the regeneration types. For both needle age-classes, A(n) and V(max) of layers were 25 and 40% higher, respectively, than the corresponding values for seedlings. The higher values of A(n), V(max) and foliar N concentration of layers compared with seedlings after two growing seasons may be associated with the larger root systems of the layers compared with the transplanted seedlings. PMID- 12651460 TI - Leaf photosynthetic characteristics of beech (Fagus sylvatica) saplings during three years of exposure to elevated CO(2) concentration. AB - Beech (Fagus sylvatica L.) seedlings were cultivated from seeds sown in pots or directly in the ground in outdoor chambers that were transparent to solar radiation, and provided either ambient air or CO(2)-enriched air (ambient + 350 &mgr;mol mol(-1)). The rooting volume was high in all experiments. In the short term experiment, potted plants were assigned to a factorial CO(2) x nutrient treatment (optimal nutrient supply and severe nutrient shortage) for 1 year. In the long-term experiment, plants were grown directly in the ground and received an optimal supply of water and nutrients in both CO(2) treatments for 3 years. Nutrient stress caused carboxylation capacity (V(m)) to decrease in the potted seedlings exposed to CO(2)-enriched air during their first growing season. In the long-term experiment with optimal nutrient supply, CO(2)-enriched air did not affect V(m), but caused an upward acclimation of maximum electron transport rate (J(m)). Consequently, there was a 14% increase in the J(m)/V(m) ratio, indicating nitrogen reallocation to maintain an equilibrium between RuBP consumption and RuBP regeneration. Both V(m) and J(m) decreased during the growing season in both CO(2) treatments. Although upward acclimation of J(m) was no longer apparent at the end of the third growing season, plants in CO(2)-enriched air maintained a higher J(m)/V(m) ratio than plants in ambient air, indicating that photosynthetic acclimation always occurred. Second flush leaves appeared during each growing season. When expressed on the basis of foliar nitrogen concentration, their photosynthetic characteristics (V(m) and J(m)) were enhanced compared with other leaves. Because the number of second flush leaves was also increased in the elevated CO(2) treatment, this response should be taken into account when modeling the effects of elevated CO(2) concentration on canopy photosynthesis. Stomatal conductance decreased in response to atmospheric CO(2) enrichment; however, the stomatal response to irradiance followed a single relationship based on two stomatal conductance models. PMID- 12651461 TI - Chlorophyll fluorescence and CO(2) assimilation of black spruce seedlings following frost in different temperature and light conditions. AB - Effects of artificial frosts on light-saturated photosynthesis (A(max)) and ground, maximal and variable fluorescence variables (F(o), F(m), and F(v) and F(v)/F(m)) were monitored on 1-year-old foliage of black spruce seedlings (Picea mariana (Mill.) BSP) grown at high (25 degrees C), moderate (15 degrees C) and low (5 degrees C) temperatures and moderate (240 &mgr;mol m(-2) s(-1)) and low (80 &mgr;mol m(-2) s(-1)) irradiances. Photoinhibition of 1-year-old foliage was greater in seedlings grown in moderate light than in seedlings grown in low light. Photoinhibition increased with decreasing growth chamber temperature at both irradiances. Most changes in F(v)/F(m) were caused by changes in F(v). Exposure to -4 degrees C decreased both F(v)/F(m) and A(max) compared with control values. The effect of the -4 degrees C frost treatment was greater in seedlings grown in low light than in seedlings grown in moderate light, probably because seedlings grown in moderate light were already partially photoinhibited before the frost treatment. Following -4 degrees C treatment, neither F(v)/F(m) nor A(max) recovered in seedlings grown in low light. Light-saturated photosynthesis decreased with decreasing growth chamber temperature. Light saturated photosynthesis was more sensitive to the -3 and -4 degrees C frost treatments in seedlings grown at 25 degrees C than in seedlings grown at 15 and 5 degrees C. The A(max) of seedlings grown at 15 degrees C was sensitive only to the -4 degrees C frost treatment, whereas A(max) of seedlings grown at 5 degrees C was not sensitive to any of the frost treatments. Recovery of A(max) following frost took longer in seedlings grown at high temperatures than in seedlings grown at low temperatures. For seedlings grown at the same temperature but under different irradiances, both A(max) and F(v)/F(m) reflected damage to the photosynthetic system following a moderate frost. However, for seedlings grown at the same irradiance but different temperatures, A(max) provided a more sensitive indicator of frost damage to the photosynthetic system than F(v)/F(m) ratio. PMID- 12651462 TI - A comparison of root growth dynamics of silver maple and flowering dogwood in compacted soil at differing soil water contents. AB - Many bottomland tree species are tolerant of compacted soil and perform well in urban environments; however, the mechanism underlying this tolerance is unknown. Increased soil water content has been shown to alleviate some of the effects of soil compaction on plant growth, presumably because increasing soil water reduces soil strength. We hypothesized that tree species tolerant of very wet soils would have opportunities for root growth in compacted soil when high soil water contents reduced soil strength, whereas species intolerant of bottomland conditions would not. We tested this hypothesis on flowering dogwood (Cornus florida L.), a mesic species intolerant of inundation, and silver maple (Acer saccharinum L.), a bottomland species. Seedlings of both species were grown in pots for 21 and 30 days, respectively, in a growth chamber in native loam soil maintained at various combinations of soil strength and soil water tension. Downward root growth rate decreased in response to increasing soil strength in both species. At low soil strength (0.6 MPa), downward root growth rate of dogwood seedlings slowed when soil was either excessively wet or dry, whereas root growth rate of silver maple seedlings increased linearly with soil water content. In moderately compacted soil (1.5 g cm(-3) bulk density), silver maple seedlings had greater root growth rate, root length per plant, and ratio of root length to root dry weight in wet soil (0.006 MPa soil water tension) than in moist and dry soils (0.026 and 0.06 MPa, respectively), even though mean oxygen diffusion rate (ODR) was only 0.28 &mgr;g cm(-2) (SE = 0.05). No such effect was detected in highly compacted soil (1.7 g cm(-3) bulk density) in either species. Mean ODR showed a weak positive correlation with soil water tension (r = 0.40, P = 0.07), but was unrelated to soil strength. We conclude that silver maple roots can grow in moderately compacted soil when high soil water content decreases soil strength, whereas dogwood is unable to take advantage of this opportunity. PMID- 12651463 TI - Kinetics of nitrogen uptake by Populus tremuloides in relation to atmospheric CO(2) and soil nitrogen availability. AB - Sustained increases in plant production in response to elevated atmospheric carbon dioxide (CO(2)) concentration may be constrained by the availability of soil nitrogen (N). However, it is possible that plants will respond to N limitation at elevated CO(2) concentration by increasing the specific N uptake capacity of their roots. To explore this possibility, we examined the kinetics of (15)NH(4) (+) and (15)NO(3) (-) uptake by excised roots of Populus tremuloides Michx. grown in ambient and twice-ambient CO(2) concentrations, and in soils of low- and high-N availability. Elevated CO(2) concentration had no effect on either NH(4) (+) or NO(3) (-) uptake, whereas high-N availability decreased the capacity of roots to take up both NH(4) (+) and NO(3) (-). The maximal rate of NH(4) (+) uptake decreased from 12 to 8 &mgr;mol g(-1) h(-1), and K(m) increased from 49 to 162 &mgr;mol l(-1), from low to high soil N availability.Because NO(3) (-) uptake exhibited mixedkinetics over the concentration range we used (10-500 &mgr;mol l( -1)), it was not possible to calculate V(max) and K(m). Instead, we used an uptake rate of 100 &mgr;mol g(-1) h(-1) as our metric of NO(3) (-) uptake capacity, which averaged 0.45 and 0.23 &mgr;mol g(-1) h(-1) at low- and high-N availability, respectively. The proximal mechanisms for decreased N uptake capacity at high-N availability appeared to be an increase in fine-root carbohydrate status and a decrease in fine-root N concentration. Both NH(4) (+) and NO(3) (-) uptake were inversely related to fine-root N concentration, and positively related to fine-root total nonstructural carbohydrate concentration. We conclude that soil N availability, through its effects on fine-root N and carbohydrate status, has a much greater influence on the specific uptake capacity of P. tremuloides fine roots than elevated atmospheric CO(2). In elevated atmospheric CO(2), changes in N acquisition by P. tremuloides appeared to be driven by changes in root architecture and biomass, rather than by changes in the amount or activity of N-uptake enzymes. PMID- 12651464 TI - Distribution of leaf mass per unit area and leaf nitrogen concentration determine partitioning of leaf nitrogen within tree canopies. AB - Distribution of leaf nitrogen with respect to leaf mass per unit area (M(a)), nitrogen per unit mass (N(m)) and nitrogen per unit area (N(a)) within peach (Prunus persica L.) tree canopies was studied in two field experiments. In one experiment, leaf light exposure and M(a) were measured on leaves from different canopy positions of peach trees subjected to five nitrogen (N) fertilization treatments. Leaf light exposure and M(a) were linearly related and the relationship was independent of N fertilization. In a subsequent experiment, N fertilizer was applied to previously unfertilized trees in midsummer, after shoot growth had terminated. Application of N fertilizer did not affect mean canopy M(a). Fertilization increased N(m) of all leaves throughout the canopy compared with non-fertilized trees. No significant relationship between N(m) and M(a) was found in either fertilized or control trees. There was a linear relationship between N(a) and M(a) and the slope of the relationship was increased by N fertilizer application. We conclude that distribution of N(a) in peach tree canopies is primarily a function of M(a) partitioning with light and N(m), which is related to soil N availability. PMID- 12651465 TI - Adjustments in leaf water relations of mangrove (Avicennia germinans) seedlings grown in a salinity gradient. AB - We used pressure-volume analysis and dewpoint hygrometry to determine leaf water relation parameters of mangrove (Avicennia germinans L.) seedlings grown at salinities of 0, 8, 20 and 32 per thousand. Seedlings responded to an increase in salinity from 0 to 32 per thousand by an increase in leaf succulence as reflected in an increase in leaf water content per unit area from 300 to 360 g m(-2). Additionally, osmotic potential at full turgor decreased from -2.3 to -3.5 MPa and osmotic potential at zero turgor decreased from -2.7 to -4.3 MPa. Cell elasticity decreased as salinity increased from 0 to 32 per thousand, as indicated by a progressive increase in volumetric modulus of elasticity from 19 to 27 MPa. Increased leaf succulence enabled leaves to sequester large amounts of solutes without adversely increasing cell osmotic pressure. On the other hand, osmotic adjustment facilitated turgor maintenance as water potential diminished. Salinity-induced decreases in tissue elasticity generated greater water potential differences between leaves and soil under saline conditions than under non-saline conditions. PMID- 12651466 TI - European larch and eastern white pine respond similarly during three years of partial defoliation. AB - To test whether trees with different leaf life spans respond differently to defoliation, eastern white pine (Pinus strobus L.) and European larch (Larix decidua Mill.) trees (9 years old in 1991) were partially defoliated by hand between July 1 and 10 in 1989, 1990 and 1991. At the end of 1991, trees of both species had received either 0, 1, 2 or 3 years of defoliation. Trees that received only 1 year of defoliation were defoliated in 1989. Variables measured included photosynthesis, twig water potential, leaf mass per area and leaf nitrogen concentration. There were few significant responses to defoliation in any of the three years of treatment in either species, and only the current-year defoliation treatments caused significant responses. Both species had reduced photosynthetic rates and less negative twig water potentials in response to defoliation in 1989. In 1990 and 1991, the defoliation treatments had no significant effect on any of the parameters measured in European larch. In 1990, there was a significant reduction in foliar nitrogen concentration in eastern white pine in response to defoliation in 1990. In 1991, eastern white pine had significantly less negative twig water potentials in response to defoliation in 1991. Leaf mass per area was not affected by defoliation in either species. We conclude that, for European larch and eastern white pine, differences in leaf life span have no effect on leaf- and twig-level responses to defoliation. PMID- 12651467 TI - Effects of season, needle age and elevated atmospheric CO(2) on photosynthesis in Scots pine (Pinus sylvestris). AB - Five-year-old Scots pine (Pinus sylvestris L.) seedlings were grown in open-top chambers at ambient and elevated (ambient + 400 &mgr;mol mol(-1)) CO(2) concentrations. Net photosynthesis (A), specific leaf area (SLA) and concentrations of nitrogen (N), carbon (C), soluble sugars, starch and chlorophyll were measured in current-year and 1-year-old needles during the second year of CO(2) enrichment. The elevated CO(2) treatment stimulated photosynthetic rates when measured at the growth CO(2) concentration, but decreased photosynthetic capacity compared with the ambient CO(2) treatment. Acclimation to elevated CO(2) involved decreases in carboxylation efficiency and RuBP regeneration capacity. Compared with the ambient CO(2) treatment, elevated CO(2) reduced light-saturated photosynthesis (when measured at 350 &mgr;mol mol( 1) in both treatments) by 18 and 23% (averaged over the growing season) in current-year and 1-year-old needles, respectively. We observed significant interactive effects of CO(2) treatment, needle age and time during the growing season on photosynthesis. Large seasonal variations in photosynthetic parameters were attributed to changes in needle chemistry, needle structure and feedbacks governed by whole-plant growth dynamics. Down-regulation of photosynthesis was probably a result of reduced N concentration on an area basis, although a downward shift in the relationship between photosynthetic parameters and N was also observed. PMID- 12651468 TI - Water utilization, plant hydraulic properties and xylem vulnerability in three contrasting coffee (Coffea arabica) cultivars. AB - Water use, hydraulic properties and xylem vulnerability to cavitation were studied in the coffee (Coffea arabica L.) cultivars San Ramon, Yellow Caturra and Typica growing in the field under similar environmental conditions. The cultivars differed in growth habit, crown morphology and total leaf surface area. Sap flow, stomatal conductance (g(s)), crown conductance (g(c)), apparent hydraulic conductance of the soil-leaf pathway (G(t)), leaf water potential (Psi(L)) and xylem vulnerability to loss of hydraulic conductivity were assessed under well watered conditions and during a 21-day period when irrigation was withheld. Sap flow, g(c), and G(t) were greatest in Typica both with and without irrigation, lowest in San Ramon, which was relatively unresponsive to the withholding of irrigation, and intermediate in Yellow Caturra. The cultivars had similar g(s) when well watered, but withholding water decreased g(s) more in Typica and Yellow Caturra than in San Ramon. Typica had substantially lower Psi(L) near the end of the unirrigated period than the other cultivars (-2.5 versus -1.8 MPa), consistent with the relatively high sap flow in this cultivar. Xylem vulnerability curves indicated that Typica was less susceptible to loss of hydraulic conductivity than the other cultivars, consistent with the more negative Psi(L) values of Typica in the field during the period of low soil water availability. During soil drying, water use declined linearly with relative conductivity loss predicted from vulnerability curves. However, cultivar-specific relationships between water use and predicted conductivity loss were not observed because of pronounced hysteresis during recovery of water use following soil water recharge. All cultivars shared the same functional relationship between integrated daily sap flow and G(t), but they had different operating ranges. The three cultivars also shared common functional relationships between hydraulic architecture and water use despite consistent differences in water use under irrigated and dry soil conditions. We conclude that hydraulic architectural traits, rate of water use per plant and crown architecture are important determinants of short- and long-term variations in the water balance of Coffea arabica. PMID- 12651469 TI - Observations of evapotranspiration in a break of slope plantation susceptible to periodic drought stress. AB - Break of slope (BOS) plantations are advocated as a means of water table control in areas where groundwater flows through colluvial deposits overlying low permeability bedrock. It is also believed that BOS plantations can supplement their water use requirements by exploiting shallow groundwater at the breaks in topographic slope. We compared measurements of BOS plantation and pasture evapotranspiration during spring, when the weather was warm and soils moist, and late summer when drought conditions prevailed. Microlysimeters and ventilated chambers were used to determine pasture and plantation floor evaporation, and heat pulse sensors were used to determine transpiration of the plantation. In spring, pasture evapotranspiration was 65% of that of the plantation, whereas, in summer, pasture evapotranspiration was equivalent to only 35% of that of the plantation. Rainfall interception by the canopy of the plantation was twice that of the pasture, reinforcing the notion that trees can help reduce groundwater recharge and alleviate dryland salinity and waterlogging. During the summer drought period, daily plantation transpiration was only 20% of that measured during spring, suggesting that the plantation was not utilizing groundwater supplies but was instead drawing from soil water supplies. This hypothesis was supported by the comparison of relative abundances of the isotopes of water ((2)H and (18)O) in soil and wood samples. We conclude that the BOS plantation is not behaving in the manner predicted, and our findings raise doubts about the predicted advantages of establishing plantations in break of slope positions. PMID- 12651470 TI - Photosynthetic capacity in a central Amazonian rain forest. AB - The vertical profile in leaf photosynthetic capacity was investigated in a terra firme rain forest in central Amazonia. Measurements of photosynthesis were made on leaves at five levels in the canopy, and a model was fitted to describe photosynthetic capacity for each level. In addition, vertical profiles of photosynthetic photon flux density, leaf nitrogen concentration and specific leaf area were measured. The derived parameters for maximum rate of electron transport (J(max)) and maximum rate of carboxylation by Rubisco (V(cmax)) increased significantly with canopy height (P < 0.05). The highest J(max) for a single canopy level was measured at the penultimate canopy level (20 m) and was 103.9 &mgr;mol m(-2) s(-1) +/- 24.2 (SE). The highest V(cmax) per canopy height was recorded at the top canopy level (24 m) and was 42.8 +/- 5.9 &mgr;mol m(-2) s( 1). Values of J(max) and V(cmax) at ground level were 35.8 +/- 3.3 and 20.5 +/- 1.3 &mgr;mol m(-2) s(-1), espectively. The increase in photosynthetic capacity with increasing canopy height was strongly correlated with leaf nitrogen concentration when examined on a leaf area basis, but was only weakly correlated on a mass basis. The correlation on an area basis can be largely explained by the concomitant decrease in specific leaf area with increasing height. Apparent daytime leaf respiration, on an area basis, also increased significantly with canopy height (P < 0.05). We conclude that canopy photosynthetic capacity can be represented as an average vertical profile, perturbations of which may be explained by variations in the environmental variables driving photosynthesis. PMID- 12651471 TI - Rapid freezing injury in red spruce: seasonal changes in sensitivity and effects of temperature range. AB - On calm, cold days in winter, sun-exposed needles of red spruce (Picea rubens Sarg.) may warm 10 to 20 degrees C above ambient air temperature, and undergo rapid (>/= 1 degrees C min(-1)) fluctuations in temperature as light breezes or passing clouds alter the energy balance of the foliage. It has been proposed that the resulting rapid freeze-thaw cycles (freezing stress) cause a type of winter injury in montane red spruce that is characterized by necrosis of sun-exposed foliage. In autumn and winter, we monitored rapid freezing stress response of needle sections from 10 montane red spruce trees by subjecting needles to rapid freezing over the temperature span typically recorded in the field. In autumn, experimental rapid freezing stress produced severe injury only at temperatures considerably lower than expected for that time of year. In winter, rapid freezing caused occasional, moderate injury in fully hardened foliage of trees susceptible to both slow and rapid freezing. Seasonal changes in sensitivity to rapid and slow freezing were correlated, suggesting that environmental factors that are known to affect sensitivity to slow freezing may also affect sensitivity to rapid freezing. Experimental manipulation of the start and end temperatures of rapid freezing stress events showed that moderate to severe needle injury can occur in susceptible trees at temperature spans slightly more extreme than those typically recorded in the field. The extent of injury was similar at different starting temperatures if rapid freezing occurred over the same temperature span. Year-old foliage was consistently less sensitive to rapid freezing stress than current year foliage, but some year-old foliage was damaged when the rapid freezing stress regime caused severe injury in current-year foliage. We conclude that rapid freeze-thaw cycles can explain light to moderate injury of current-year foliage, but they do not explain the more severe and widespread pattern of foliar damage that has occurred intermittently over at least the last 18 years. PMID- 12651472 TI - Use of a single-tree simulation model to predict effects of ozone and drought on growth of a white fir tree. AB - A physiologically based, single-tree simulation model, TREGRO, was parameterized with existing phenological, allometric, and growth data and used to predict effects of ozone and drought on growth of a 53-year-old white fir (Abies concolor (Gord. & Glend.) Lindl. ex Hildebr.) tree following a 3-year model simulation. Multiple experimental simulations were conducted to assess the individual and interactive effects of ozone (O(3)) exposure and drought on growth of white fir. The effects of O(3) were imposed as reductions in carbon (C) assimilation of 0, 2.5, 5, 10, and 20%. Drought was imposed as 0, 10, 25, and 50% reductions in total annual precipitation. The results of the simulations were compared with the effects of O(3) on white fir seedlings grown in the presence and absence of ozone in open-top chambers and with a field survey of white fir trees subjected to a gradient of O(3). In the O(3) simulations, an O(3)-induced reduction in C assimilation of 2.5% reduced total tree biomass and branch total nonstructural carbohydrate (TNC) content by < 7%. Although quantifiable in simulation experiments, such small reductions would probably not be detectable in the field. Results from both an open-top chamber experiment and a field survey indicated that reductions in C assimilation of white fir growing in elevated O(3) were much greater than 2.5%, but were not statistically different from control values. A simulated O(3) reduction in C assimilation of >/= 10% reduced total tree biomass by 7% and branch TNC by 55%. Results from the field survey indicated that branch elongation was reduced in response to increased O(3) concentration, corroborating the simulated response of reduced C allocation to the branches of white fir. Although simulated reductions in total annual precipitation of >/= 25% reduced final tree biomass, the simulated reductions also reduced O(3) uptake and therefore reduced the O(3) response of white fir. However, a combination of low amounts of O(3) (2.5% reduction in C assimilation) and drought (25% reduction in annual precipitation) synergistically reduced C gain of white fir more than either stress individually. Our simulations predict that moderate drought (no more than a 25% reduction in total annual precipitation) may not ameliorate the response of white fir to O(3) and that moderate amounts of atmospheric O(3) and drought could be more detrimental to white fir than either stress singly. PMID- 12651473 TI - Allometric differences between current-year shoots and large branches of deciduous broad-leaved tree species. AB - Current-year shoots are mostly made of primary tissues, whereas first-order branches comprise mainly secondary tissues. Differences in tissue composition of these units reflect differences in functional design. We compared the allometry of current-year shoots and first-order branches in eight deciduous broad-leaved tree species and examined the functional differences underlying the design of current-year shoots and first-order branches. Allometric relationships of first order branches tended to be compatible with predictions of the pipe model and elastic similarity model. That is, allometric constants of the relationships between leaf mass and stem diameter at the branch base and between stem diameter and stem mass were 2.0 and 0.33-0.38, respectively, indicating that the functional regulation of stem form of first-order branches can be predicted by the two models. However, allometric relationships of current-year shoots were not compatible with the predictions of the pipe and elastic similarity models. Thus, the allometric constant of the relationship between leaf mass and stem diameter at the base of current-year shoots was larger than 2.0, and the allometric constant of the relationship between stem length and stem diameter of shoots was larger than 1.0 in all species examined. However, current-year shoots had an allometric constant of leaf mass against stem length that was less than 1.0, suggesting a functional demand on shoot design to reduce self-shading. Also, allometric constants of stem length against stem diameter at the shoot base were larger in monopodial species than in sympodial species, whereas allometric constants of leaf mass per shoot against stem length were smaller in monopodial species than in sympodial species. We propose that the allometries of current year shoots reflect their function as disposable units for temporary leaf arrangement. PMID- 12651474 TI - Photosynthesis in Norway spruce seedlings infected by the needle rust Chrysomyxa rhododendri. AB - Chrysomyxa rhododendri (DC.) De Bary is a needle rust with a host shift between Rhododendron sp. and Norway spruce (Picea abies (L.) Karst.), penetrating only the new developing flushes of the conifer. Because little is known about its effects on trees, we investigated several parameters related to photosynthesis in artificially infected 3-year-old Norway spruce seedlings. The potential efficiency of photosystem II (PSII; derived from chlorophyll fluorescence measurements) was reduced in infected current-year needles as soon as disease symptoms were visible, about three weeks after inoculation. Two weeks later, photosynthetic O(2) evolution (P(max)) of infected needles was less than 20% of control needles, whereas respiratory O(2) uptake (R(D)) was about three times higher than that of control needles. Nonstructural carbohydrate concentrations were about 60% of control values in all parts of the shoots of infected trees. Photosynthetic inhibition was associated with marked decreases in chlorophyll concentration and chlorophyll a/b ratio but only a small reduction in carotenoid concentration. In infected trees, P(max) of noninfected 1-year-old and 2-year-old needles was 50 and 80% higher than in the corresponding age class of needles of control trees. Estimation of potential daily net dry mass production, based on P(max), R(D), specific leaf area, carbon content and needle biomass, indicated that seedlings infected once were able to produce 60%, and those infected twice only 25%, of the dry mass of controls. We conclude that afforestation and regeneration of Norway spruce is seriously impaired in regions where seedlings are frequently attacked by Chrysomyxa. PMID- 12651475 TI - Acid mist and soil Ca and Al alter the mineral nutrition and physiology of red spruce. AB - We examined the effects and potential interactions of acid mist and soil solution Ca and Al treatments on foliar cation concentrations, membrane-associated Ca (mCa), ion leaching, growth, carbon exchange, and cold tolerance of red spruce (Picea rubens Sarg.) saplings. Soil solution Ca additions increased foliar Ca and Zn concentrations, and increased rates of respiration early in the growing season (July). Soil Al treatment had a broad impact, reducing foliar concentrations of Ca, Mg, Mn, P and Zn, and resulting in smaller stem diameters, sapling heights and shoot lengths compared with soil treatments with no added Al. Aluminum treatment also reduced respiration when shoots were elongating in July and decreased net photosynthesis at the end of the growing season (September). Three lines of evidence suggest that Al-induced alterations in growth and physiology were independent of foliar Ca status: (1) Ca concentrations in foliage of Al treated saplings were within the range of sufficiency established for red spruce; (2) mCa concentrations were unaffected by Al treatment; and (3) no Al x Ca interactions were detected. Acid mist treatment increased foliar Fe and K concentrations and increased leaching of Ca, Mg, Mn, Zn, Fe, and Al from foliage. Leaching losses of Ca were more than twice those of the element with the next highest amount of leaching (Zn), and probably led to the reductions in mCa concentration and membrane stability of acid-treated saplings. Acidic mist resulted in enhanced shoot growth, and consistent reductions in foliar cold tolerance in the fall and winter. Of the few significant interactions among treatments, most involved the influence of mist pH and Al treatment on foliar nutrition. In general, reductions in cation concentration associated with Al addition were greater for pH 5.0-treated saplings than for pH 3.0-treated saplings. We propose that H(+)-induced leaching of mCa from mesophyll cells is the mechanism underlying acid-induced reductions in foliar cold tolerance of red spruce. PMID- 12651476 TI - Acclimation of photosynthesis and respiration to simulated climatic warming in northern and southern populations of Acer saccharum: laboratory and field evidence. AB - Physiological acclimation and genotypic adaptation to prevailing temperatures may influence forest responses to future climatic warming. We examined photosynthetic and respiratory responses of sugar maple (Acer saccharum Marsh.) from two portions of the species' range for evidence of both phenomena in a laboratory study with seedlings. A field study was also conducted to assess the impacts of temperature acclimation on saplings subjected to an imposed temperature manipulation (4 degrees C above ambient temperature). The two seedling populations exhibited more evidence of physiological acclimation to warming than of ecotypic adaptation, although respiration was less sensitive to short-term warming in the southern population than in the northern population. In both seedling populations, thermal compensation increased photosynthesis by 14% and decreased respiration by 10% in the warm-acclimated groups. Saplings growing in open-top field chambers at ambient temperature and 4 degrees C above ambient temperature showed evidence of temperature acclimation, but photosynthesis did not increase in response to the 4 degrees C warming. On the contrary, photosynthetic rates measured at the prevailing chamber temperature throughout three growing seasons were similar, or lower (12% lower on average) in saplings maintained at 4 degrees C above ambient temperature compared with saplings maintained at ambient temperature. However, the long-term photosynthetic temperature optimum for saplings in the field experiment was higher than it was for seedlings in either the 27 or the 31 degrees C growth chamber. Respiratory acclimation was also evident in the saplings in the field chambers. Saplings had similar rates of respiration in both temperature treatments, and respiration showed little dependence on prevailing temperature during the growing season. We conclude that photosynthesis and respiration in sugar maple have the potential for physiological acclimation to temperature, but exhibit a low degree of genetic adaptation. Some of the potential for acclimation to a 4 degrees C increase above a background of naturally fluctuating temperatures may be offset by differences in water relations, and, in the long term, may be obscured by the inherent variability in rates under field conditions. Nevertheless, physiologically based models should incorporate seasonal acclimation to temperature and permit ecotypic differences to influence model outcomes for those species with high genetic differentiation between regions. PMID- 12651477 TI - Tree water use and rainfall partitioning in a mature poplar-pasture system. AB - Traditionally, poplars (Populus) have been planted to control erosion on New Zealand's hill-slopes, because of their capacity to dry out and bind together the soil, by reducing effective rainfall and increasing evapotranspiration and soil strength. However, the effect of widely spaced poplars on the partitioning of soil water and rainfall has not been reported. This study determined rainfall partitioning for 18 mid-spring days in a mature P. deltoides (Bart. ex Marsh, Clone I78)-pasture association (37 stems per hectare, unevenly spaced at 16.4 +/- 0.4 m) and compared it with a traditional open pasture system in grazed areas of a hill environment. Tree transpiration was measured by the heat pulse technique. A time-driven mathematical model was used to set a zero offset, adjust anomalous values and describe simultaneous sap velocity time courses of trees. The model showed that daylight sap flow velocities can be represented with a nonlinear Beta function (R(2) > 0.98), and differences in the parameters representing the initiation, duration and conformation of the sap velocity can be tested statistically to discern tree transpiration differences during the day. Evapotranspiration was greater for the poplar-pasture association than for the open pasture (2.7-3.0 versus 2.2 mm day(-1)). The tree canopy alone contributed 0.92 mm day(-1) as transpiration and 1.37 mm day(-1) as interception, whereas evapotranspiration of the pasture understory was only 0.4-0.6 mm day(-1). Despite the higher water use of the poplar-pasture association, soil water in the 0-300 mm soil stratum was higher than, or similar to, that of the open pasture. Tree shading decreased evapotranspiration and pasture accumulation under the trees. PMID- 12651478 TI - Frost hardening of Pinus radiata seedlings: effects of temperature on relative growth rate, carbon balance and carbohydrate concentration. AB - Pinus radiata (D. Don) seedlings were grown for 100 days at day/night temperatures of 10/1, 15/1, 20/1 and 25/1 degrees C, to determine whether temperatures above a threshold of 5 degrees C influence frost hardiness development. Relationships between hardening and relative growth rate, carbohydrate concentration and net carbon balance were also investigated. Seedlings hardened at a nearly constant rate in each treatment, although the rate of hardening was strongly temperature dependent. It increased as the temperature declined, but in a curvilinear fashion. Temperatures below 9.5 degrees C were effective in hardening the seedlings. During the daily temperature cycle, dehardening occurred at temperatures above the threshold, whereas hardening occurred at temperatures below the threshold. The net difference between the two processes determined the development of frost hardiness. The development of frost hardiness was negatively correlated with relative growth rate and positively correlated with the accumulation of starch and sugars. We conclude that frost hardening is a complex process that is causally linked to carbohydrate concentrations. PMID- 12651479 TI - Modeling daily gas exchange of a Douglas-fir forest: comparison of three stomatal conductance models with and without a soil water stress function. AB - Modeling stomatal conductance is a key element in predicting tree growth and water use at the stand scale. We compared three commonly used models of stomatal conductance, the Jarvis-Loustau, Ball-Berry and Leuning models, for their suitability for incorporating soil water stress into their formulation, and for their performance in modeling forest ecosystem fluxes. We optimized the parameters of each of the three models with sap flow and soil water content data. The optimized Ball-Berry model showed clear relationships with air temperature and soil water content, whereas the optimized Leuning and Jarvis-Loustau models only showed a relationship with soil water content. We conclude that use of relative humidity instead of vapor pressure deficit, as in the Ball-Berry model, is not suitable for modeling daily gas exchange in Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) in the Speulderbos forest near the village of Garderen, The Netherlands. Based on the calculated responses to soil water content, we linked a model of forest growth, FORGRO, with a model of soil water, SWIF, to obtain a forest water-balance model that satisfactorily simulated carbon and water (transpiration) fluxes and soil water contents in the Douglas-fir forest for 1995. PMID- 12651480 TI - Dynamics of symbiotic establishment between an IAA-overproducing mutant of the ectomycorrhizal fungus Hebeloma cylindrosporum and Pinus pinaster. AB - To clarify the early steps of symbiotic establishment, we studied the dynamics of Pinus pinaster (Ait.) Sol. tap root colonization and mycorrhiza formation by an IAA-overproducing mutant of the ectomycorrhizal fungus Hebeloma cylindrosporum Romagnesi and by the corresponding wild type strain. Differences between wild type and mutant strains were quantitative rather than qualitative and were detected two days after inoculation. Both fungal strains established a typical Hartig net when they colonized the tap roots. Consequently, colonized tap roots exhibited features of a true mycorrhiza and fungal colonization enhanced plant growth. Fungal colonization and Hartig net formation were more rapid with the mutant than with the wild type. Colonization, especially with the mutant strain, increased rhizogenesis and the production of mycorrhizas. The mutant formed a hypertrophic Hartig net in tap roots and mycorrhizal short roots and we obtained evidence that the process of short root transformation into mycorrhiza started before their emergence from the tap root. Hyphae of the Hartig net in the tap root penetrated the cortex of young lateral roots at the beginning of their elongation, after the endodermis layer broke under the pressure of the elongating lateral root. Colonization was inhibited when triiodobenzoic acid was added to the culture medium, providing circumstantial evidence that auxin is involved in mycorrhiza formation. PMID- 12651481 TI - Energy dissipation in drought-avoiding and drought-tolerant tree species at midday during the Mediterranean summer. AB - Photosynthetic performance was monitored during two consecutive summers in four co-occurring evergreen Mediterranean tree species growing on a south-facing rocky slope. In response to midday water stress, the drought-avoiding species Pinus halepensis Mill. exhibited marked stomatal closure (g(s)) but no changes in stem water potential (Psi(s)), whereas the drought-tolerant species Quercus coccifera L., Q. ilex ssp. ballota (Desf.) Samp. and Juniperus phoenicea L. displayed declines in midday g(s) and Psi(s). The higher resistance to CO(2) influx in needles of P. halepensis compared with the other species did not result in either a proportional increase in non-radiative dissipation of excess energy or photo inactivation of photosystem II (PSII). No significant differences were found among species either in the de-epoxidation state of the xanthophyll cycle (DPS) or in the pool of its components on a total chlorophyll basis (VAZ). Despite contrasting midday assimilation rates, the three drought-tolerant species all exhibited a pronounced drop in photochemical efficiency at midday that was characterized by a decrease in the excitation capture efficiency of the open PSII centers. Although photoinhibition was not fully reversed before dawn, it apparently did not result in cumulative photo-damage. Thus, the drought-avoiding and drought-tolerant species employed different mechanisms for coping with excess light during the midday depression in photosynthesis that involved contrasting midday photochemical efficiencies of PSII and different degrees of dynamic photoinhibition as a photo-protective mechanism. These behaviors may be related to the different mechanisms employed by drought-avoiding and drought-tolerant species to withstand water deficit. PMID- 12651482 TI - Direct effect of elevated CO(2) on nocturnal in situ leaf respiration in nine temperate deciduous tree species is small. AB - Direct (i.e., short-term) effects of elevated CO(2) on nocturnal in situ leaf respiration rate were measured in nine deciduous tree species (seven genera) in 20 3.5-4.0-h experiments. During the experiments, CO(2) concentration was alternated between 400 and 800 ppm (approximately 40 and 80 Pa of CO(2)). Data analysis accounted for effects on respiration rate of the normal decline in temperature with time after sunset. The median response to a 40-Pa increase in CO(2) was a 1.5% decrease in respiration rate, with responses ranging from a 5.6% inhibition to a 0.4% stimulation. Direct effects of elevated CO(2) on respiration were similar among the species. Thus, the response of nocturnal leaf respiration rate to a short-term CO(2) increase was small, and of little practical importance to the accuracy of measurements of respiration involving similar changes in CO(2) concentration during measurement. These direct respiratory responses of leaves to elevated CO(2) would translate into only slight, if any, effects on the carbon balance of temperate deciduous forests in a future atmosphere containing as much as 80 Pa CO(2). PMID- 12651483 TI - Carbon isotope discrimination and oxygen isotope composition in clones of the F(1) hybrid between slash pine and Caribbean pine in relation to tree growth, water-use efficiency and foliar nutrient concentration. AB - The objectives of this study were: (1) to examine how foliar carbon isotope discrimination (Delta) and oxygen isotope composition (delta(18)O) are related to tree growth, ash mineral nutrient concentration and foliar nutrient concentration in 7-year-old clones of the F(1) hybrid between slash pine (Pinus elliottii Engelm.) and Caribbean pine (P. caribaea var. hondurensis Barr. et Golf.) in subtropical Australia; and (2) to evaluate the potential of using foliar Delta, ash mineral nutrient concentration and delta(18)O measurements for selecting F(1) hybrid pine clones with high water-use efficiency (WUE) and growth potential. There were significant differences in tree growth, foliar Delta, delta(18)O and ash mineral nutrient concentration among the eight clones tested. Significant negative linear relationships existed between tree growth and Delta, extrapolating to zero growth at Delta = 24-30 per thousand. There were strong genetic correlations (r = -0.83 to -0.96) between Delta and tree growth, particularly tree height. Significant non-genetic correlations (r = -0.62 to 0.80) existed between Delta and foliar K concentration. Foliar delta(18)O, ash mineral nutrient concentration and foliar nutrient concentration were unrelated to tree growth. In the F(1) hybrid pine clones, variation in tree WUE, as reflected by Delta, was largely attributed to a genetic effect on leaf photosynthetic capacity rather than on stomatal conductance, as reflected by foliar delta(18)O. PMID- 12651484 TI - Dry season conditions determine wet season water use in the wet-tropical savannas of northern Australia. AB - Daily and seasonal patterns of transpiration were measured in evergreen eucalypt trees growing at a wet (Darwin), intermediate (Katherine) and dry site (Newcastle Waters) along a steep rainfall gradient in a north Australian savanna. Relationships between tree size and tree water use were also determined. Diameter at breast height (DBH) was an excellent predictor of sapwood area in the five eucalypt species sampled along the rainfall gradient. A single relationship existed for all species at all sites. Mean daily water use was also correlated to DBH in both wet and dry seasons. There were no significant differences in the relationship between DBH and tree water use at Darwin or Katherine. Among the sites, tree water use was lowest at Newcastle Waters at all DBHs. The relationship between DBH and tree leaf area was similar between species and locations, but the slope of the relationship was less at the end of the dry season than at the end of the wet season at all locations. There was a strong relationship between sapwood area and leaf area that was similar at all sites along the gradient. Transpiration rates were significantly lower in trees at the driest site than at the other sites, but there were no significant differences in transpiration rates between trees growing at Darwin and Katherine. Transpiration rates did not vary significantly between seasons at any site. At all sites, there was only a 10% decline in water use per tree between the wet and dry seasons. A monthly aridity index (pan evaporation/rainfall) and predawn leaf water potential showed strong seasonal patterns. It is proposed that dry season conditions exert control on tree water use during the wet season, possibly through an effect on xylem structure. PMID- 12651485 TI - Selection of white spruce families in the context of climate change: heat tolerance. AB - To assess the responses and plasticity of white spruce seedlings (Picea glauca (Moench) Voss) to high temperatures, 12 open-pollinated families differing in growth performance were exposed to a 30-min heat treatment of 42, 44, 46, 48, or 50 degrees C with or without heat preconditioning at 38 degrees C for 5 h. Damage was evaluated based on chlorophyll fluorescence parameters after heat preconditioning, after the heat treatments and during a 7-day recovery period. Visible needle damage was also evaluated after the heat treatments and 14 days later. Chlorophyll fluorescence parameters indicated that seedlings subjected to a heat treatment of 42-43 degrees C lost the ability to phosphorylate and donate water to photosystem II (PSII). A heat treatment of 44-46 degrees C severely limited the ability of the seedlings to use NADPH and ATP in the Calvin cycle. Based on visible needle damage, families with superior height-growth performance were more sensitive to heat stress than families with intermediate or inferior height-growth performance. Moreover, families with superior height-growth performance had low photochemical efficiencies in the light (DeltaF/F(m)') after heat treatment. Heat preconditioning increased the thermotolerance of the seedlings. However, the data suggest that white spruce seedlings exhibiting fast growing characteristics under present conditions may not grow as well at higher temperatures. PMID- 12651486 TI - Effects of drought preconditioning on thermotolerance of photosystem II and susceptibility of photosynthesis to heat stress in cedar seedlings. AB - Changes in photosystem II (PSII) thermotolerance during drought and recovery were studied under controlled conditions in three Mediterranean cedar species (Cedrus brevifolia Henry, C. libani Loudon and C. atlantica Manetti). The temperature at which the quantum yield of PSII photochemistry was reduced by 15% of its value at 25 degrees C was 3 to 4 degrees C higher in drought-treated plants than in well watered plants. The drought-induced increase in PSII thermotolerance was already evident 8 days after water had been withheld from the seedlings, when net CO(2) assimilation was still at 80% of its initial value, and was visible for up to 12 days after re-watering. When seedlings of the three species were exposed to temperatures above 45 degrees C for 5 h, both maximal quantum yield of PSII photochemistry and net CO(2) assimilation rate were significantly reduced in unconditioned seedlings, whereas drought-preconditioned seedlings were almost unaffected by the heat treatment. Drought-preconditioned seedlings still exhibited a higher tolerance to heat stress than unconditioned seedlings 60 days after re-watering, although the transient, drought-induced osmotic adjustment had fully disappeared. Among species, C. atlantica was the most heat sensitive, whereas the heat treatment had no significant effect on the parameters measured in C. brevifolia. PMID- 12651487 TI - Reinvigoration of mature chestnut (Castanea sativa) by repeated graftings and micropropagation. AB - Gradual reinvigoration of adult chestnut (Castanea sativa M. cv. Montemarano) shoots was obtained by serial grafting onto juvenile rootstocks. The phenomenon was evaluated on the basis of percentage of primary nodes regenerating axillary shoots and length and number of shoots (> 10 mm) per primary node. In vitro growth of explants from serially grafted shoots was significantly lower than that of explants from seedlings at the end of the establishment phase. Only microshoots from seedlings and plants that had been serially grafted four times could be subcultured on proliferation medium. Repeated subculture on medium containing a low cytokinin concentration induced progressive reinvigoration of microshoots derived from plants that had been serially grafted four times. The number of axillary shoots per explant increased significantly after six subcultures. After 12 subcultures, microshoots from serially grafted plants showed an increase in stem elongation, rooting and plantlet survival. After in vitro stabilization, there was no difference in in vitro performance between microshoots derived from seedlings and serially grafted plants. Microshoots multiplied from serially grafted plants displayed only a transitory appearance of juvenile traits. PMID- 12651488 TI - Photosynthetic nitrogen-use efficiency in evergreen broad-leaved woody species coexisting in a warm-temperate forest. AB - Photosynthetic nitrogen-use efficiency (PNUE, photosynthetic capacity per unit leaf nitrogen) varies among species from different habitats and correlates with several ecological characteristics such as leaf life span and leaf mass per area. We investigated eight evergreen broad-leaved woody species with different leaf life spans that coexist in a warm-temperate forest. We determined photosynthetic capacity at ambient CO(2) concentration in saturated light, nitrogen concentration, and the concentration of ribulose-1,5-bisphosphate carboxylase (RuBPCase), a key enzyme of photosynthesis and the largest sink of nitrogen in leaves. Each species showed a strong correlation between photosynthetic capacity and RuBPCase concentration, and between RuBPCase concentration and nitrogen concentration. Photosynthetic capacity of leaves decreased with increasing leaf life span, whereas PNUE did not correlate significantly with leaf life span. There was a twofold variation in PNUE among species. This relatively small variation in PNUE is consistent with the argument that species that coexist in a single habitat maintain a similar PNUE. The two components of PNUE-photosynthetic rate per unit RuBPCase and RuBPCase per unit leaf nitrogen-were not significantly correlated with other leaf characteristics such as leaf life span and leaf mass per area. We conclude that differences in PNUE are relatively small among coexisting species and that differences in absolute amounts of photosynthetic proteins lead to differences in photosynthetic productivity among species. PMID- 12651489 TI - Photosynthetic utilization efficiency of absorbed photosynthetically active radiation by Scots pine and birch forest stands in the southern Taiga. AB - Absorption and utilization of photosynthetically active radiation (PAR) were investigated in Scots pine (Pinus sylvestris L.) and birch (Betula pendula Roth.) stands that were 41 years old at the end of the experimental period. Canopy depth of the Scots pine stand was about half that of the birch stand (6.5 versus 11.0 m), but absorption of PAR was similar in the two stands. The Scots pine forest canopy, with a leaf area index of 8.9, absorbed 90% of the incoming PAR (APAR), whereas the birch forest canopy, with a leaf area index of 5.9, absorbed 92% of APAR. During maximum foliage development, the upper Scots pine canopy absorbed more PAR than the upper birch canopy (75 versus 66%). The upper, middle and lower layers of the Scots pine canopy contained 37, 48 and 15% of the total needle surface area, respectively. The corresponding distribution of foliage surface area in the three layers of the birch canopy was 50, 30 and 20%, respectively. Measurements of photosynthetic rate were combined with estimates of leaf area index and stand phytomass to determine rates of primary production on a sunny day, a cloudy day, and on an annual basis. The energy equivalents of short- and long-term carbon gain were used with determinations of APAR to calculate photosynthetic utilization efficiency. Throughout the growing season, photosynthetic utilization efficiency of APAR in the upper canopy layer of the Scots pine forest was almost twice that in the lower canopy layer. In the birch forest, photosynthetic utilization efficiency was greater in the lower canopy layer than in the upper canopy layer. In all cases, utilization efficiency was higher in the birch stand than in the Scots pine stand (52 versus 29 J kJ(-1)). Taking account of respiration of the non-photosynthetic parts of each stand (night respiration of needles or leaves; respiration of branches, trunk and roots), estimated utilization efficiency of APAR for net primary production was 11 J kJ(-1) for Scots pine and 19 J kJ(-1) for birch. Solar conversion ratios, expressed as whole-plant net primary productivity per unit of APAR for the growing season, were 0.81 g MJ(-1) for Scots pine and 0.93 g MJ(-1) for birch. PMID- 12651490 TI - Radial variation in sap flow in five laurel forest tree species in Tenerife, Canary Islands. AB - Variations in radial patterns of xylem water content and sap flow rate were measured in five laurel forest tree species (Laurus azorica (Seub.) Franco, Persea indica (L.) Spreng., Myrica faya Ait., Erica arborea L. and Ilex perado Ait. ssp. platyphylla (Webb & Berth.) Tutin) growing in an experimental plot at Agua Garcia, Tenerife, Canary Islands. Measurements were performed around midday during warm and sunny days by the heat field deformation method. In all species, water content was almost constant (around 35% by volume) over the whole xylem cross-sectional area. There were no differences in wood color over the whole cross-sectional area of the stem in most species with the exception of E. arborea, whose wood became darker in the inner layers. Radial patterns of sap flow were highly variable and did not show clear relationships with tree diameter or species. Sap flow occurred over the whole xylem cross-sectional area in some species, whereas it was limited to the outer xylem layers in others. Sap flow rate was either similar along the xylem radius or exhibited a peak in the outer part of the xylem area. Low sap flow rates with little variation in radial pattern were typical for shaded suppressed trees, whereas dominant trees exhibited high sap flow rates with a peak in the radial pattern. Stem damage resulted in a significant decrease in sap flow rate in the outer xylem layers. The outer xylem is more important for whole tree water supply than the inner xylem because of its larger size. We conclude that measurement of radial flow pattern provides a reliable method of integrating sap flow from individual measuring points to the whole tree. PMID- 12651491 TI - Leaf water relations and stomatal behavior of four allopatric Eucalyptus species planted in Mediterranean southwestern Australia. AB - In 1986, four allopatric Eucalyptus species (E. camaldulensis Dehnh, E. saligna Smith, E. leucoxylon F. Muell and E. platypus Hook.) were planted together in a 480-mm rainfall zone, in 8-m wide contour belts as part of a plan to minimize waterlogging and secondary salinization. Throughout 1997, 1998 and 1999, there was significant inter-specific variation in predawn leaf water potential (Psi(pd)); however, maximum stomatal conductance (g(sm)) only differed significantly between species in mid to late summer. Relationships between g(sm) and Psi(pd) were significant and showed that stomata of E. camaldulensis were significantly more sensitive to Psi(pd), and presumably soil water potential, than stomata of E. leucoxylon or E. platypus. When applied to the Psi(pd) data, these relationships predicted that g(sm), and by inference transpiration, varied much less between species than Psi(pd). Diurnal measurements throughout the season confirmed this prediction, and showed that E. camaldulensis and E. saligna avoided drought by gaining access to deeper water, whereas E. leucoxylon and E. platypus maintained greater g(sm) at a given water stress than E. camaldulensis or E. saligna. Osmotic potentials measured after rehydration and water release curves of the leaves indicated that different mechanisms accounted for the apparent drought tolerance of E. leucoxylon and E. platypus. In summer, E. leucoxylon reduced osmotic potential at full and zero turgor by similar amounts compared with winter. In summer, E. platypus had a significantly lower bulk elastic modulus and relative water content at turgor loss point than E. camaldulensis, E. saligna or E. leucoxylon. This elastic adjustment resulted in a larger difference between osmotic potential at full and zero turgor in summer than in winter. The inherently low osmotic potential in E. leucoxylon and elastic adjustment in E. platypus resulted in turgor loss at a similar and significantly lower water potential than in E. camaldulensis or E. saligna. These results have implications for species selection for planting to manage groundwater recharge in areas prone to waterlogging and secondary salinization. PMID- 12651492 TI - Comparative water relations of mature mahogany (Swietenia macrophylla) trees with and without lianas in a subhumid, seasonally dry forest in Bolivia. AB - Many evergreen mahogany (Swietenia macrophylla King) trees in the seasonally dry Bajo Paragua forest in northeast Bolivia carry substantial liana loads. Evergreen lianas may impede the growth of their host trees in various ways, including competition for water. Hypotheses tested were that water relations status differs (a) between lianas and their host trees, and (b) between trees infested with lianas that were cut 3.5 months previously (treated trees) and control trees with intact lianas. Diurnal measurements of stomatal conductance (g(s)) and leaf water potential (Psi) were made on canopy leaves of treated and control trees and lianas at the start and end of the dry season. Lianas had higher (less negative) Psi values (mean and predawn) and higher diurnal g(s) (expressed as mean or sum of diurnal values) than mahogany trees, indicating that lianas had a higher demand for, and ability to obtain, water than their host trees. Control and treated trees had a similar water relations status, suggesting that removal of lianas had no effect on the water relations of the trees, even at the end of the dry season. We conclude that either both life forms have conservative water relations that were unaffected by water availability in our study, or that the trees and lianas have access to sufficient and different sources of water because of differences in their rooting depths. Our data are consistent with studies of temperate species, indicating that lianas do not interfere with water availability to their host trees. PMID- 12651493 TI - Predicting spring phenology and frost damage risk of Betula spp. under climatic warming: a comparison of two models. AB - Timing of bud burst and frost damage risk for leaves of Betula spp. in response to climatic warming in Finland was examined with two models. In the first model, ontogenetic development in spring was triggered by an accumulation of chilling temperatures. The second model assumed an additional signal from the light climate. The two models gave radically different estimates of frost damage risk in response to climate warming. The chilling-triggered model forecast a significant and increasing risk with increased warming, whereas the light-climate triggered model predicted little or no risk. The chilling-triggered model is widely applied in phenological research; however, there is increasing experimental evidence that light conditions play a role in the timing of spring phenology. Although it is not clear if the light response mechanisms are appropriately represented in our model, the results imply that reliance on a light signal for spring development would afford a degree of protection against possible frost damage under climate warming that would not be present if chilling were the sole determinant. Further experimental tests are required to ascertain the light-related mechanisms controlling phenological timing, so that credible model extrapolations can be undertaken. PMID- 12651494 TI - High efficiency genetic transformation of sour orange (Citrus aurantium) and production of transgenic trees containing the coat protein gene of citrus tristeza virus. AB - In preliminary experiments on Agrobacterium-mediated transformation of citrus, we found transformation events occurring in callus formed from the cambium. Factors affecting Agrobacterium-sour orange (Citrus aurantium L.) interactions, such as culture medium, explant source and culture conditions, were studied to assess competence for transformation in such callus and to improve transformation frequency. Cell divisions and redifferentiation from the transgenic cells leading to transformation events were stimulated more by a combination of benzylaminopurine (BAP) + naphthalene-acetic acid (NAA) in the regeneration selection medium than by BAP alone. Both age and source of the sour orange plant material affected transformation frequency. Explants from 4-month-old seedlings grown in the greenhouse showed higher transformation frequency than younger and older plant materials, indicating that they had a more suitable balance between dedifferentiated cells competent for transformation and Agrobacterium virulence. Enhancement of transformation frequency enabled us to incorporate the coat protein gene of citrus tristeza virus (CTV) in a sufficient number of sour orange plants to be able to evaluate this strategy for producing CTV-resistant plants. PMID- 12651495 TI - Decreased needle longevity of fertilized Douglas-fir and grand fir in the northern Rockies. AB - Changes in nutrient availability significantly affect canopy dynamics in conifers. To elucidate these effects, we experimentally fertilized mixed conifer stands at several sites across the northern Rocky Mountains. We measured needle longevity, total branch length and foliated length along the main branch axis, and determined mean retained cohort length on mid-canopy branches of shade intolerant Douglas-fir (Pseudotsuga menziesii var. glauca (Beissn.) Franco) and shade-tolerant grand fir (Abies grandis Lindl.). Needle longevity ranged from 6 to 8 years in Douglas-fir and from 7 to 8 years in grand fir on unfertilized plots. Fertilization significantly decreased needle longevity by 26 and 27% in Douglas-fir and grand fir, respectively. However, the foliated branch length remained unchanged following fertilization and was similar for both species, indicating a 33% increase in mean branch length per needle cohort in Douglas-fir and a 27% increase in grand fir. These data are consistent with the theory that foliated branch length and needle longevity are a result of the ecological light compensation point (ELCP), which links the inherent physiology of the leaf with the availability of resources in the leaf environment. Mid-canopy ELCP was approximately 74 and 71 cm from the branch terminus in Douglas-fir and grand fir, respectively, regardless of fertilization. We hypothesize that fertilization enhanced needle production and annual shoot growth resulted in a higher rate of shading of older needles. The shaded needles were unable to maintain a positive carbon balance and abscised. The results demonstrate that foliated branch length of Douglas-fir and grand fir in the northern Rocky Mountains can be treated as a homeostatic response to fertilization, whereas foliar turnover is plastic. PMID- 12651496 TI - Reduction of isoprene emissions from live oak (Quercus fusiformis) with oak wilt. AB - Many plants emit isoprene, a hydrocarbon that has important influences on atmospheric chemistry. Pathogens may affect isoprene fluxes, both through damage to plant tissue and by changing the abundance of isoprene-emitting species. Live oaks (Quercus fusiformis (Small) Sarg. and Q. virginiana Mill) are major emitters of isoprene in the southern United States, and oak populations in Texas are being dramatically reduced by oak wilt, a widespread fungal vascular disease. We investigated the effects of oak wilt on isoprene emissions from live oak leaves (Q. fusiformis) in the field, as a first step in exploring the physiological effects of oak wilt on isoprene production and the implications of these effects for larger-scale isoprene fluxes. Isoprene emission rates per unit dry leaf mass were 44% lower for actively symptomatic leaves than for leaves on healthy trees (P = 0.033). Isoprene fluxes were significantly negatively correlated with rankings of disease activity in the host tree (fluxes in leaves on healthy trees > healthy leaves on survivor trees > healthy leaves on the same branch as symptomatic leaves > symptomatic leaves; isoprene per unit dry mass: Spearman's rho = -0.781, P = 0.001; isoprene per unit leaf area: Spearman's rho = -0.652, P = 0.008). Photosynthesis and stomatal conductance were reduced by 57 and 63%, respectively, in symptomatic relative to healthy leaves (P < 0.05); these reductions were proportionally greater than the reductions in isoprene emissions. Low isoprene emission rates in symptomatic leaves are most simply explained by physiological constraints on isoprene production, such as water stress as a result of xylem blockage, rather than direct effects of the oak wilt fungus on isoprene synthesis. The effects of oak wilt on leaf-level isoprene emission rates are probably less important for regional isoprene fluxes than the reduction in oak leaf area across landscapes. PMID- 12651497 TI - Physiological responses of radiata pine roots to soil strength and soil water deficit. AB - We investigated physiological responses of radiata pine (Pinus radiata D. Don) roots to soil strength and soil water deficit by measuring the osmotic potential (Psi(pi)) and yield turgor (Y) in the elongation zone of root segments of seedlings growing (i) in polyethylene glycol 4000-containing rooting solution of different water potentials (Psi(s)) and (ii) in soil of different soil strengths (Q) at the same soil matric potential (Psi(m)). Root elongation rate (Deltal/Deltat) decreased progressively with decreasing Psi(s) and was associated with decreased Psi(pi) and decreased turgor pressure (P). Osmotic adjustment occurred at Psi(s) < -0.2 MPa. Over a range in Psi(s) of -0.01 to -1.0 MPa, Psi(pi) fell 0.3 MPa whereas P fell 0.7 MPa. Mean Psi in the solution experiment was 0.37 MPa and did not differ significantly with Psi(s) (P = 0.10). Root elongation rate decreased exponentially as Q increased from 0 to 3.0 MPa, and was associated with an increase in P of 0.11 MPa as a consequence of Psi(pi) decreasing by the same amount. Mean Y in the soil experiment was 0.49 MPa and did not change significantly with Q (P = 0.87). PMID- 12651498 TI - Structure-function relationships during secondary phloem development in an angiosperm tree, Aesculus hippocastanum: microtubules and cell walls. AB - We studied the dynamics of the cortical microtubule (CMT) cytoskeleton during differentiation of axial secondary phloem elements in taproots and epicotyls of Aesculus hippocastanum L. (horse-chestnut) saplings. Indirect immunofluorescence microscopy of alpha-tubulin and transmission electron microscopy revealed that fusiform cambial cells possessed a reticulum of CMTs in which individual microtubules were randomly arranged. During differentiation of these cambial cell derivatives into secondary phloem cells, the CMTs were rearranged to become helically oriented, regardless of phloem cell type. Although helical CMTs were a persistent feature of all axial elements of the secondary phloem (sieve elements, companion cells, phloem parenchyma, and fiber-sclereids), some modifications of this arrangement occurred as cells differentiated. Thus, at late stages of cell differentiation, sieve elements possessed nearly transverse CMTs, pronounced bundling of CMTs was seen in phloem parenchyma, and the density of CMTs in the helical arrays of fibers increased markedly. Additionally, phloem parenchyma possessed rings of CMTs in association with developing pit areas. Aspects of the development and chemistry of cell walls were also examined during phloem cytodifferentiation. PMID- 12651499 TI - Quantifying stomatal and non-stomatal limitations to carbon assimilation resulting from leaf aging and drought in mature deciduous tree species. AB - Gas exchange techniques were used to investigate light-saturated carbon assimilation and its stomatal and non-stomatal limitations over two seasons in mature trees of five species in a closed deciduous forest. Stomatal and non stomatal contributions to decreases in assimilation resulting from leaf age and drought were quantified relative to the maximum rates obtained early in the season at optimal soil water contents. Although carbon assimilation, stomatal conductance and photosynthetic capacity (V(cmax)) decreased with leaf age, decreases in V(cmax) accounted for about 75% of the leaf-age related reduction in light-saturated assimilation rates, with a secondary role for stomatal conductance (around 25%). However, when considered independently from leaf age, the drought response was dominated by stomatal limitations, accounting for about 75% of the total limitation. Some of the analytical difficulties associated with computing limitation partitioning are discussed, including path dependence, patchy stomatal closure and diffusion in the mesophyll. Although these considerations may introduce errors in our estimates, our analysis establishes some reasonable boundaries on relative limitations and shows differences between drought and non-drought years. Estimating seasonal limitations under natural conditions, as shown in this study, provides a useful basis for comparing limitation processes between years and species. PMID- 12651500 TI - Temporal variability in basal isoprene emission factor. AB - Seasonal variability in basal isoprene emission factor (&mgr;g C g(-1) h(-1) or nmol m(-2) s(-1), leaf temperature at 30 degrees C and photosynthetically active radiation (PAR) at 1000 &mgr;mol m(-2) s(-1)) was studied during the 1998 growing season at Duke Forest in the North Carolina Piedmont. Emissions from eight upper canopy white oak (Quercus alba L.) leaves were measured periodically from the onset of isoprene emission on Day of Year (DOY) 119 (April 29) to leaf senescence in late October (DOY 299). Emissions from four leaves were measured under basal conditions with a controlled-environment cuvette system equipped with 10-ml gas tight syringes and a reduction gas detector. Emissions from the other four leaves were measured under ambient conditions with the same system. Emission rates from the four leaves measured under ambient conditions were adjusted to basal conditions based on the PAR and leaf temperature algorithms of Guenther et al. (1993). The seasonal onset of isoprene emission was in agreement with previous studies where cumulative degree days from the date of the last spring frost were used to estimate bud break, leaf expansion, and increase in basal emission factor (EF). Between DOY 141 (May 21) and 240 (August 28), mean meteorological conditions 6 to 18 h prior to the EF measurements (ambient PAR and temperature) explained up to 78% of the variability in mean basal EF between measurement periods. Summertime mean isoprene emission potential was reached on DOY 141 (May 21) and was maintained until DOY 240 (August 28), when isoprene emission began to decline monotonically as leaf senescence approached. The mean value for leaves measured under ambient conditions and adjusted to basal conditions for DOY 141 240 was 75.6 &mgr;g C g(-1) h(-1) (74.2-79.1), whereas the mean value for leaves measured under basal conditions was 72.9 &mgr;g C g(-1) h(-1) (64.7-88.9). Between DOY 141 and 240, daily mean isoprene EFs varied from 54 to 96 &mgr;g C g( 1) h(-1) (27 to 49 nmol m(-2) s(-1)). In agreement with previous work at this and other sites, basal isoprene emission rates of fully exposed leaves at the crown apex of this tree were about 20% higher than those of the selected leaves. The length of the period prior to measurement of isoprene emission, during which meteorology was correlated with basal EF, appeared to be related to the timing and periodicity of meteorological change, and probably explains quantitative differences in the length of this period among studies. The empirical equation that we derived for this effect explained variability in midday EFs at the study site, but its general applicability remains to be tested. Strong diurnal changes in EF (as high as a factor of 2) are implied in this study, and should be examined further. PMID- 12651501 TI - Carbon and nitrogen mobilization in Larix x eurolepis leafy stem cuttings assessed by dual (13)C and (15)N labeling: relationships with rooting. AB - Changes in use of both stored and newly synthesized sources of carbon (C) and nitrogen (N) were investigated during rooting of leafy cuttings of Larix x eurolepis A. Henry. We used dual (13)C and (15)N long-term labeling of reserves of stock plants and followed isotope dilution of the labels in the cuttings to determine the respective proportions of C and N derived from stock plant reserves (Q(C,old), Q(N,old)) and from newly synthesized sources (Q(C,new), Q(N,new)). We also assessed their partitioning into the growing needles and roots. Because rooting development occurred over several months, destructive harvests were made during three periods (t(0): April 9; t(1): May 15 to June 5; t(2): June 12 to June 26). Total C content (Q(C)) of cuttings did not change with time, whereas total N content (Q(N)) decreased significantly between t(0) and t(2). In rooted cuttings, unlike unrooted cuttings, total N concentration decreased less markedly in needles and in the lower stem component between t(1) and t(2) than between t(0) and t(2). The lower stem of rooted cuttings contained more Q(N,old) than that of unrooted cuttings and showed lower C:N and (13)C:(15)N ratios. The increase in Q(N,new) in the lower stem was positively correlated with rooting status. At t(1), new roots appeared and grew mainly at the expense of Q(C,new) and Q(N,old). By t(2), root growth was mainly dependent on new assimilates. We conclude that in leafy cuttings the initial amount of N reserves, rather than C reserves, may constitute a limiting factor for rooting. PMID- 12651502 TI - Leaf orientation, light interception and stomatal conductance of Eucalyptus globulus ssp. globulus leaves. AB - Juvenile and adult leaves of the heteroblastic species Eucalyptus globulus Labill. ssp. globulus did not show active diurnal orientation toward or away from incident radiation. Juvenile leaves of a late-maturing sapling of a Tasmanian provenance were evenly distributed in all azimuth sectors. In contrast, an early maturing sapling of the same age from Wilsons Promontory, Victoria had a predominance of adult leaf blades facing east and west. Mid-vein and blade angles of juvenile and adult leaves were non-random with an overall vertical declination of the leaves. Both leaf types intercepted a greater irradiance during the morning than at midday. Sub-horizontal juvenile leaves intercepted 22% more irradiance than vertical adult leaves during the middle of the day. The amphistomatal and isobilateral adult leaves intercepted sunlight equally on both leaf surfaces. Stomatal conductance was variable during the period of measurement but was similar for the Tasmanian juvenile (0.4 to 0.9 cm s(-1)) leaves and Wilsons Promontory adult (0.5 to 1.2 cm s(-1)) leaves. Greater light interception by the sub-horizontal juvenile leaves would confer a growth advantage to saplings and regenerating canopies. Reduced light interception and leaf temperature of vertical adult E. globulus leaves would assist in water conservation, particularly at high solar angles. PMID- 12651503 TI - Coarse root elongation rate estimates for interior Douglas-fir. AB - Accurate estimates of root growth rates are important for root system modeling, and the spread of root systems may be an important determinant of belowground site occupancy. Estimating root system growth rates is complicated because missing, discontinuous, and false annual growth rings make root cross sections difficult to age. These irregularities can occur even in roots of dominant conifers with rare or absent stem growth ring abnormalities. Incomplete rings were noted in the root growth rings of nine co-dominant interior Douglas-fir (Pseudotsuga menziesii var. glauca (Beissn.) Franco) trees. Coarse root (> 1.0-cm diameter) elongation rates were estimated by fitting a geometric mean regression line to ring count and lateral distance data. In all nine roots examined, the geometric mean regression slope was well within the range of the 95% confidence interval for the ordinary least squares regression of lateral distance versus age, suggesting that measurement error may have been negligible. Coarse root elongation rates (which ranged from 2.8 to 15.3 cm year(-1) and averaged 7.4 cm year(-1)) in the interior Douglas-fir trees studied were much lower than those reported by others. This discrepancy may be a result of limited soil water availability, soil heterogeneity (both soil water content and soil texture were highly variable across short distances) and fragmentation of belowground growing space. PMID- 12651504 TI - Influence of water table decline on growth allocation and endogenous gibberellins in black cottonwood. AB - Cottonwoods occur in riparian areas where water table depth generally varies with the elevation of the adjacent river. Plant adaptation to the riparian zone requires the coordination of root elongation to maintain contact with the water table during the summer decline. We investigated the effects of rate of water decline on growth allocation and concentrations of endogenous gibberellins (GAs) in black cottonwood (Populus trichocarpa Torr. & A. Gray ex Hook.) saplings. Rhizopods were used to achieve water decline rates of 0, 2 and 4 cm day(-1). Root elongation approximately doubled in response to the 2 cm day(-1) treatment, whereas leaf area was reduced. A water decline rate of 4 cm day(-1) led to water stress, as evidenced by reduced growth, increased leaf diffusive resistance, decreased water potential, and leaf senescence and abscission. Endogenous GAs were extracted, purified and analyzed by gas chromatography-selected ion monitoring with internal [(2)H(2)]GA standards. Across the sampled plant organs, GAs were generally highest in shoot tips and sequentially lower in basal stems, root tips, leaves and upper roots; GAs were thus abundant in rapidly growing tissues. Of the GAs measured, GA(1) tended to predominate, followed sequentially by GA(3), GA(8), GA(19), GA(20), GA(29) and GA(4). There was little relationship between GA concentration and growth allocation across the water table decline treatments, although GA(8) was consistently reduced in plants experiencing water table decline. Because GA(8) is the final gibberellin in the metabolic sequence, it might be useful for assessing historic patterns of GAs and growth rate. This study demonstrated changes in growth allocation in response to water table decline, but provided little evidence that endogenous GAs play a primary role in the regulation of root elongation in response to water table decline. PMID- 12651505 TI - Genetic and environmental control of seasonal carbohydrate dynamics in trees of diverse Pinus sylvestris populations. AB - We explored environmental and genetic factors affecting seasonal dynamics of starch and soluble nonstructural carbohydrates in needle and twig cohorts and roots of Scots pine (Pinus sylvestris L.) trees of six populations originating between 49 degrees and 60 degrees N, and grown under common garden conditions in western Poland. Trees of each population were sampled once or twice per month over a 3-year period from age 15 to 17 years. Based on similarity in starch concentration patterns in needles, two distinct groups of populations were identified; one comprised northern populations from Sweden and Russia (59-60 degrees N), and another comprised central European populations from Latvia, Poland, Germany and France (49-56 degrees N). Needle starch concentrations of northern populations started to decline in late spring and reached minimum values earlier than those of central populations. For all populations, starch accumulation in spring started when minimum air temperature permanently exceeded 0 degrees C. Starch accumulation peaked before bud break and was highest in 1 year-old needles, averaging 9-13% of dry mass. Soluble carbohydrate concentrations were lowest in spring and summer and highest in autumn and winter. There were no differences among populations in seasonal pattern of soluble carbohydrate concentrations. Averaged across all populations, needle soluble carbohydrate concentrations increased from about 4% of needle dry mass in developing current-year needles, to about 9% in 1- and 2-year-old needles. Root carbohydrate concentration exhibited a bimodal pattern with peaks in spring and autumn. Northern populations had higher concentrations of fine-root starch in spring and autumn than central populations. Late-summer carbohydrate accumulation in roots started only after depletion of starch in needles and woody shoots. We conclude that Scots pine carbohydrate dynamics depend partially on inherited properties that are probably related to phenology of root and shoot growth. PMID- 12651506 TI - Introduction to BOREAS special issue. AB - Carbon flux attributed to the boreal forest biome significantly impacts the global carbon cycle. The multi-scale Boreal Ecosystem Atmosphere Study (BOREAS) was initiated to investigate exchanges between boreal forests and the atmosphere, to examine the effects of increased atmospheric CO(2) concentration on boreal forests, to develop and improve process models to describe exchanges between boreal forests and the atmosphere, and to develop methods for the large-scale application of these models. Field measurements have shown that boreal ecosystems differ significantly from their temperate and tropical counterparts with respect to seasonality, productivity and biosphere-atmosphere interactions. PMID- 12651507 TI - Measuring and modeling conductances of black spruce at three organizational scales: shoot, branch and canopy. AB - To investigate the extent to which the energy balance of a globally important ecosystem is controlled by biological and environmental processes, measurements of water vapor flux were made on individual black spruce (Picea mariana [Mill.] B.S.P.) shoots, branches, and a whole canopy at the BOREAS Southern Study Area Old Black Spruce (SSA OBS) site. These measurements were used to estimate stomatal, branch boundary layer and canopy boundary layer conductances to water vapor. On a projected needle area basis, stomatal conductances varied between 14 and 92 mmol m(-2) s(-1), and total branch conductance varied seasonally between zero and about 35 mmol m(-2) s(-1). On a ground area basis, total canopy conductance varied between 24 and 105 mmol m(-2) s(-1). Total canopy conductance was partitioned into aerodynamic and physiological components by using shoot scale measurements scaled by leaf area index. Good agreement was found with an independent estimate of aerodynamic conductance measured when the canopy was wet. Compared with most coniferous forests, the canopy was relatively uncoupled from the atmosphere, and at the ecosystem scale, the control of water vapor flux was approximately equally divided between physiological and abiotic conductances. Two widely used steady-state models of stomatal conductance were parameterized from the shoot and branch measurements. Parameters varied considerably throughout the growing season. A time-constant term was added to these static models to construct dynamic models of stomatal conductance under naturally varying environmental conditions. The dynamic versions of the models outperformed the static versions in explaining stomatal response to rapidly changing environmental conditions. The length of the time-constant term, derived using the dynamic models, suggested that stomata were slow to respond to changing environmental conditions, and that the speed of the response was strongly temperature dependent. PMID- 12651508 TI - Responses of trembling aspen and hazelnut to vapor pressure deficit in a boreal deciduous forest. AB - The branch bag method was used to monitor photosynthesis and transpiration of trembling aspen (Populus tremuloides Michx.) and hazelnut (Corylus cornuta Marsh.) over a 42-day midsummer period in 1996, as part of the Boreal Ecosystem Atmosphere Study (BOREAS). During the same period, daytime measurements of stomatal conductance (g(s)) and leaf water potential (Psi(leaf)) were made on these species, and sap flow was monitored in aspen stems by the heat pulse method. Weather conditions during the study period were similar to the long-term average. Despite moist soils, both species showed an inverse relationship between daytime g(s) and vapor pressure deficit (D) when D was > 0.5 kPa. Daytime Psi(leaf) was below -2 MPa in aspen and near -1.5 MPa in hazelnut, except on rainy days. These results are consistent with the hypothesis that stomatal responses are constrained by hydraulic resistance from root to leaf, and by the need to maintain Psi(leaf) above a minimum threshold value. Reductions in g(s) on sunny afternoons with elevated ambient D (maximum 2.3 kPa) were associated with a significant decrease in photosynthetic rates. However, day-to-day variation in mean carbon assimilation rate was small in both species, and appeared to be governed more by solar radiation than D. These results may be generally applicable to healthy aspen stands under normal midsummer conditions in the southern boreal forest. However, strong reductions in carbon uptake may be expected at the more extreme values of D (> 4 kPa) that occur during periods of regional drought, even if soil water is not locally limiting. PMID- 12651509 TI - Isoprene emission inventory for the BOREAS southern study area. AB - An isoprene emission inventory for a section of boreal forest in central Saskatchewan was developed based on measured emission rates from the two dominant isoprene-emitting species, black spruce (Picea mariana (Mill.) BSP) and aspen (Populus tremuloides Michx.). The micrometeorological gradient technique was used to determine isoprene emission factors for establishing the inventory. Isoprene fluxes were measured during each of the three BOREAS intensive field campaigns (IFCs) during the 1994 growing season. Measured isoprene fluxes varied from 0.04 to 3.3 mg C m(-2) h(-1) over the black spruce canopy, and from 0.05 to 7.3 mg C m(-2) h(-1) above the aspen forest. Midsummer standard isoprene emission fluxes were 1.2 mg C m(-2) h(-1) and 2.3 mg C m(-2) h(-1) (at 20 degrees C and photosynthetically active radiation (PAR) of 1000 &mgr;mol m(-2) s(-1)) for black spruce and aspen, respectively. With light and temperature differences accounted for, there was an apparent seasonal effect on emissions with the highest rates in the summer months. The total amount of isoprene emitted from this section of the boreal forest was estimated to be 8.6 Gg C year(-1), which is about 1% of the net ecosystem carbon exchange for the study area. Aspen was the largest contributor, accounting for approximately 70% of the total. Branch enclosure and relaxed eddy accumulation measurements made at the black spruce site were used to define the uncertainty associated with flux measurements. Emission rates obtained by the gradient, enclosure and relaxed eddy accumulation methods showed good agreement when normalized to standard light and temperature conditions. The coefficient of variance between the three techniques was 12% for summer (IFC-2) measurements. The sensitivity of the annual isoprene emission total to the assignment of mean irradiance and temperature was also examined. If the hourly mean temperatures were increased by 1 degrees C throughout the growing season, annual carbon loss due to isoprene emission would increase by 14% from 8.6 to 9.8 Gg C. PMID- 12651510 TI - Estimating CO(2) flux from snowpacks at three sites in the Rocky Mountains. AB - Soil surface CO(2) flux (F(s)) is the dominant respiratory flux in many temperate forest ecosystems. Snowpacks increase this dominance by insulating the soil against the low temperature to which aboveground components are exposed. However, measurement of F(s) in winter may be impeded by snow cover. Likewise, developing annual F(s) models is complicated by seasonal variation in root and microbial metabolism. We compared three methods of measuring sub-snow F(s): (1) dynamic chamber measurements at the upper snowpack surface (F(snow)), (2) dynamic chamber measurements at the soil surface via snowpits (F(soil)), and (3) static estimates based on measured concentrations of carbon dioxide ([CO(2)]) and conductance properties of the snowpack (F(diffusional)). Methods were compared at a mid elevation forest in northeastern Washington, a mid-elevation forest in northern Idaho, and a high-elevation forest and neighboring meadow in Wyoming. The methods that minimized snowpack disturbance, F(diffusional) and F(snow), yielded similar estimates of F(s). In contrast, F(soil) yielded rates two to three times higher than F(snow) at the forested sites, and seven times higher at the subalpine meadow. The ratio F(soil)/F(snow) increased with increasing snow depth when compared across all sites. Snow removal appears to induce elevated soil flux as a result of lateral CO(2) diffusion into the pit. We chose F(snow) as our preferred method and used it to estimate annual CO(2) fluxes. The snowpack was present for 36% of the year at this site, during which time 132 g C m(-2), or 17% of the annual flux, occurred. We conclude that snowpack CO(2) flux is quantitatively important in annual carbon budgets for these forests and that the static and dynamic methods yield similar and reasonable estimates of the flux, as long as snowpack disturbance is minimized. PMID- 12651511 TI - Use of middle infrared radiation to estimate the leaf area index of a boreal forest. AB - The leaf area index (LAI) of boreal forest can be estimated using reflected radiation recorded by satellite sensors. Measurements of visible and near infrared radiation are commonly used in the normalized difference vegetation index (NDVI) to estimate LAI. However, research, mainly in tropical forest, has demonstrated that LAI is related more closely to radiation of middle infrared wavelengths than of visible wavelengths. This paper derives a vegetation index, VI3, based on radiation from vegetation recorded at near and middle infrared wavelengths. For a boreal forest canopy, the relationship between VI3 and LAI was observed to be much stronger than that between NDVI and LAI. In addition, the LAI estimated using VI3 accounted for about 76% of the variation in field estimates of LAI, compared with about 46% when using the NDVI. We conclude that information provided by middle infrared radiation should be considered when estimating the leaf area index of boreal forest. PMID- 12651512 TI - Regional assessment of boreal forest productivity using an ecological process model and remote sensing parameter maps. AB - An ecological process model (BIOME-BGC) was used to assess boreal forest regional net primary production (NPP) and response to short-term, year-to-year weather fluctuations based on spatially explicit, land cover and biomass maps derived by radar remote sensing, as well as soil, terrain and daily weather information. Simulations were conducted at a 30-m spatial resolution, over a 1205 km(2) portion of the BOREAS Southern Study Area of central Saskatchewan, Canada, over a 3-year period (1994-1996). Simulations of NPP for the study region were spatially and temporally complex, averaging 2.2 (+/- 0.6), 1.8 (+/- 0.5) and 1.7 (+/- 0.5) Mg C ha(-1) year(-1) for 1994, 1995 and 1996, respectively. Spatial variability of NPP was strongly controlled by the amount of aboveground biomass, particularly photosynthetic leaf area, whereas biophysical differences between broadleaf deciduous and evergreen coniferous vegetation were of secondary importance. Simulations of NPP were strongly sensitive to year-to-year variations in seasonal weather patterns, which influenced the timing of spring thaw and deciduous bud burst. Reductions in annual NPP of approximately 17 and 22% for 1995 and 1996, respectively, were attributed to 3- and 5-week delays in spring thaw relative to 1994. Boreal forest stands with greater proportions of deciduous vegetation were more sensitive to the timing of spring thaw than evergreen coniferous stands. Similar relationships were found by comparing simulated snow depth records with 10-year records of aboveground NPP measurements obtained from biomass harvest plots within the BOREAS region. These results highlight the importance of sub grid scale land cover complexity in controlling boreal forest regional productivity, the dynamic response of the biome to short-term interannual climate variations, and the potential implications of climate change and other large scale disturbances. PMID- 12651513 TI - Water availability and branch length determine delta(13)C in foliage of Pinus pinaster. AB - The stable carbon isotope composition (delta(13)C) of foliage integrates signals resulting from environmental and hydraulic constraints on water movement and photosynthesis. We used branch length as a simple predictor of hydraulic constraints to water fluxes and determined the response of delta(13)C to varying water availability. Foliage up to 6 years old was taken from Pinus pinaster Ait. trees growing at four sites differing in precipitation (P; 414-984 mm year(-1)) and potential evaporation (ET; 1091-1750 mm year(-1)). Branch length was the principal determinant of temporal trends in delta(13)C. The strong relationship between delta(13)C and branch length was a function of hydraulic conductance, which was negatively correlated with branch length (r(2) = 0.84). Variation in P and ET among sites was reflected in delta(13)C, which was negatively correlated with P/ET (r(2) = 0.66). However, this analysis was confounded by differences in branch length. If the effects of branch length on delta(13)C were first removed, then the 'residual' delta(13)C was more closely related to P/ET (r(2) = 0.99), highlighting the importance of accounting for variation in hydraulic constraints to water flux between sites and years. For plant species that exhibit considerable phenotypic plasticity in response to changes in environment (e.g., variation in leaf area, branch length and number, or stem form), the environmental effects on delta(13)C in foliage can only be reliably assessed if deconvoluted from hydraulic constraints. PMID- 12651514 TI - Influence of nutrient supply on shade-sun acclimation of Picea abies seedlings: effects on foliar morphology, photosynthetic performance and growth. AB - Norway spruce seedlings (Picea abies Karst.) were grown in low light for one year, under conditions of adequate and limiting nutrition, then transferred to high light. Three months after transfer we measured photosynthesis, leaf nitrogen concentration, leaf chlorophyll concentration and leaf mass per area (LMA) of current-year and 1-year-old shoots; silhouette area ratio (SAR, the ratio of shoot silhouette area to projected needle area) was also measured in current-year shoots. At the foliage level, the effects of light and nutrient treatments differed markedly. Light availability during foliage expansion primarily affected LMA and SAR (morphological acclimation at the needle and shoot level, respectively). By contrast, nutrient supply in high light affected photosynthetic capacity per unit of leaf tissue (physiological acclimation at the cellular level) but did not affect LMA and SAR. The capacity for shade-sun acclimation in foliage formed before transfer to high light differed greatly from that of foliage formed following the transfer. The morphological inflexibility of mature needles (measured by LMA) limited their shade-sun acclimation potential. In contrast, at high nutrient supply, shoots that developed just after the change in photosynthetic photon flux density largely acclimated, both morphologically and physiologically, to the new light environment. The acclimation response of both current- and 1-year-old shoots was prevented by nutrient limitation. Analysis of growth at the whole-plant level largely confirmed the conclusions drawn at the shoot level. We conclude that nutrient shortage subsequent to the opening of a canopy gap may strongly limit the acclimation response of Norway spruce seedlings. Successful acclimation was largely related to the plant's ability to produce sun foliage and adjust whole-plant biomass allocation rapidly. PMID- 12651515 TI - Effect of pruning on nitrogen dynamics within crowns of Pinus radiata. AB - Stem injection of (15)N-labeled ammonium sulfate was used to determine effects of pruning on canopy nitrogen dynamics in open-grown Pinus radiata D. Don in New Zealand. Trees were planted in July 1990 and the isotope introduced in December 1994. Tree crowns were divided into three zones: base section, from which branches of pruned trees were removed; mid section, between the pruned zone and the height of the trees at the start of the year in which they were pruned; and top section, which grew predominantly after the isotope was applied. Pruning removed 32% of the green crown length, representing 75% of foliage biomass. Needles were sampled from each region of the crown until July 1996. Branch growth was used to predict foliage biomass for each sampling occasion. Approximately 45% of the applied isotope was recovered from needles sampled in December 1994 (1 week after application and immediately before pruning), two-thirds of which occurred in needles in the base section. Thereafter, changes in isotope content of needles in the base section of unpruned trees largely reflected foliage biomass fluctuations and dilution of the isotope by continued uptake from the unlabeled soil nitrogen pool. Recovery of isotope in needles from the mid-crown section increased by 58 and 86% from December 1994 to July 1995 in control and pruned trees, respectively. Within this crown section, there was evidence of isotope translocation from old to new needles, with both isotope dilution and efflux observed in the needle cohorts that had been present at the time the isotope was applied. Therefore, isotope dynamics did not reflect the dynamics of the total nitrogen pool in the mid-crown section. By July 1996, a small proportion of the applied isotope was recovered from the new foliage formed in the top section of the crown. Within the top section, isotope dynamics closely matched total nitrogen fluxes. Pruning the lower crown did not affect nitrogen dynamics elsewhere in the crown for the following 18 months. PMID- 12651516 TI - Characterization of Pin m III cDNA in western white pine. AB - Maximum accumulation of Pin m III protein in western white pine (Pinus monticola Dougl. ex D. Don) needles occurred during the winter months. To characterize Pin m III, an expression cDNA library from poly(A)+ mRNA of needles was immunoscreened and the full length cDNA was cloned. An open reading frame of 486 bases encodes a protein of 161 amino acid residues with a molecular mass of 18 kD and a predicted isoelectric point of 5.5. The deduced amino acid sequence had some similarities (37%) with an intracellular pathogenesis-related (PR) protein from garden asparagus (Asparagus officinalis L.) and the major pollen allergen from white birch (Betula verrucosa J. F. Ehrh.), which are members of the ribonuclease-like PR-10 family. Phylogenetic analysis provided circumstantial evidence that Pin m III may be grouped with intracellular PRs from asparagus and potato (Solanum tuberosum L.), while the allergens formed another subgroup. Northern analysis showed that the Pin m III gene was preferentially expressed during cold acclimation with the highest expression in the fall and winter months, preceding the peak of Pin m III protein accumulation. Tissue specificity expression analysis indicated that the gene was strongly expressed in roots and twigs. Higher amounts of the homologous protein (Pin l I) and its transcript accumulated in sugar pine (Pinus lambertiana Dougl.) needles infected with blister rust compared with healthy needles. PMID- 12651517 TI - Transpiration and forest structure in relation to soil waterlogging in a Hawaiian montane cloud forest. AB - Transpiration, leaf characteristics and forest structure in Metrosideros polymorpha Gaud. stands growing in East Maui, Hawaii were investigated to assess physiological limitations associated with flooding as a mechanism of reduced canopy leaf area in waterlogged sites. Whole-tree sap flow, stomatal conductance, microclimate, soil oxidation-reduction potential, stand basal area and leaf area index (LAI) were measured on moderately sloped, drained sites with closed canopies (90%) and on level, waterlogged sites with open canopies (50-60%). The LAI was measured with a new technique based on enlarged photographs of individual tree crowns and allometric relationships. Sap flow was scaled to the stand level by multiplying basal area-normalized sap flow by stand basal area. Level sites had lower soil redox potentials, lower mean stand basal area, lower LAI, and a higher degree of soil avoidance by roots than sloped sites. Foliar nutrients and leaf mass per area (LMA) in M. polymorpha were similar between level and sloped sites. Stomatal conductance was similar for M. polymorpha saplings on both sites, but decreased with increasing tree height (r(2) = 0.72; P < 0.001). Stand transpiration estimates ranged from 79 to 89% of potential evapotranspiration (PET) for sloped sites and from 28 to 51% of PET for level sites. Stand transpiration estimates were strongly correlated with LAI (r(2) = 0.96; P < 0.001). Whole-tree transpiration was lower at level sites with waterlogged soils, but was similar or higher for trees on level sites when normalized by leaf area. Trees on level sites had a smaller leaf area per stem diameter than trees on sloped sites, suggesting that soil oxygen deficiency may reduce leaf area. However, transpiration per unit leaf area did not vary substantially, so leaf level physiological behavior was conserved, regardless of differences in tree leaf area. PMID- 12651518 TI - Spatial variations in xylem sap flux density in the trunk of orchard-grown, mature mango trees under changing soil water conditions. AB - Circumferential and radial variations in xylem sap flux density in trunks of 13 year-old mango (Mangifera indica L.) trees were investigated with Granier sap flow sensor probes under limiting and non-limiting soil water conditions. Under non-limiting soil water conditions, circumferential variation was substantial, but there was no apparent relationship between sap flux density and aspect (i.e., the radial position of the sensor probes on the trunk relative to the compass). Hourly sap flux densities over 24 hours at different aspects were highly pair wise correlated. The relationships between different aspects were constant during well-watered periods but highly variable under changing soil water conditions. Sap flux density showed marked radial variation within the trunk and a substantial flux was observed at the center of the trunk. For each selected aspect on each tree, changes in sap flux densities over time at different depths were closely correlated, so flux at a particular depth could be extrapolated as a multiple of flux from 0 to 2 cm beneath the cambium. However, depth profiles of sap flux density differed between trees and even between aspects within a tree, and also varied in an unpredictable manner as soil water conditions changed. Nevertheless, over a period of non-limiting soil water conditions, depth profiles remained relatively constant. Based on the depth profiles obtained during these periods, a method is described for calculating total sap flow in a mango tree from sap flux density at 0-2 cm beneath the cambium. Total daily sap flows obtained were consistent with water use estimated from soil water balance. PMID- 12651519 TI - Seed germination of six mature neotropical rain forest species in response to dehydration. AB - We studied effects of dehydration and rehydration on germination of seeds of six mature tropical rain forest species-Cupania glabra Swartz, Cymbopetalum baillonii Fries, Poulsenia armata (Miq.) Standl., Stemmadenia donnell-smithii (Rose) Woodson, Rheedia edulis Triana & Planch. and an understory palm Chamaedorea alternans H. Wendl.-from Veracruz, Mexico. Before the seeds were sown, their water content was reduced by 0 (control), 30, 54 and 72% of their original water content. Dehydration affects the ability of seeds to rehydrate, as well as the rate and final percentage of germination when seeds are subsequently rehydrated. Seed survival and germination behavior showed three patterns: (1) C. baillonii, P. armata and S. donnell-smithii had greater tolerance to seed dehydration than C. glabra, C. alternans and R. edulis; (2) partial dehydration enhanced germinability of C. glabra and C. baillonii seeds; and (3) partial dehydration of C. alternans and R. edulis seeds resulted in delayed or sporadic germination. A relationship was found between the effects of dehydration on germination and the seasonality of seed production. PMID- 12651520 TI - Reduction in turgid water volume in jack pine, white spruce and black spruce in response to drought and paclobutrazol. AB - Significant reductions in needle water content were observed in white spruce (Picea glauca (Moench) Voss), black spruce (Picea mariana (Mill) B.S.P.), and jack pine (Pinus banksiana Lamb.) seedlings in response to a 10-day drought, although turgor was apparently maintained. When the seedlings were re-watered after the drought, jack pine needles regained their original saturated volume, whereas white spruce and black spruce needles did not. Significant drought induced reductions in turgor-loss volume (i.e., tissue volume at the point of turgor loss) were observed in shoots of all three species, especially jack pine. Repeated exposure to 7 days of drought or treatment with the cytochrome P(450) inhibitor, paclobutrazol ((2RS,3RS)-1-(4-chlorophenyl)-4,4-dimethyl-2-(1,2,4 triazol-1-yl)-pentan-3-ol), reduced seedling height relative to that of untreated controls in all three species. The reductions in saturated and turgor-loss needle volumes in the paclobutrazol-treated seedlings were comparable with those of seedlings subjected to a 10-day drought. The treatment-induced reductions in shoot and needle water contents enabled seedlings to maintain turgor with tissue volumes close to, or below, the turgor-loss volume of untreated seedlings. Paclobutrazol-treated seedlings subsequently survived drought treatments that were lethal to untreated seedlings. PMID- 12651521 TI - Differences in leaf gas exchange and water relations among species and tree sizes in an Arizona pine-oak forest. AB - We compared leaf gas exchange and water potential among the dominant tree species and major size classes of trees in an upland, pine-oak forest in northern Arizona. The study included old-growth Gambel oak (Quercus gambelii Nutt.), and sapling, pole, and old-growth ponderosa pines (Pinus ponderosa var. scopulorum Dougl. ex Laws.). Old-growth oak had higher predawn leaf water potential (Psi(leaf)) than old-growth pine, indicating greater avoidance of soil water stress by oak. Old-growth oak had higher stomatal conductance (G(w)), net photosynthetic rate (P(n)), and leaf nitrogen concentration, and lower daytime Psi(leaf) than old-growth pine. Stomatal closure started at a daytime Psi(leaf) of about -1.9 MPa for pine, whereas old-growth oak showed no obvious reduction in G(w) at Psi(leaf) values greater than -2.5 MPa. In ponderosa pine, P(n) and G(w) were highly sensitive to seasonal and diurnal variations in vapor pressure deficit (VPD), with similar sensitivity for sapling, pole, and old-growth trees. In contrast, P(n) and G(w) were less sensitive to VPD in Gambel oak than in ponderosa pine, suggesting greater tolerance of oak to atmospheric water stress. Compared with sapling pine, old-growth pine had lower morning and afternoon P(n) and G(w), predawn Psi(leaf), daytime Psi(leaf), and soil-to-leaf hydraulic conductance (K(l)), and higher foliar nitrogen concentration. Pole pine values were intermediate between sapling and old-growth pine values for morning G(w) and daytime Psi(leaf), similar to sapling pine for predawn Psi(leaf), and similar to old-growth pine for morning and afternoon P(n), afternoon G(w), K(l), and foliar nitrogen concentration. For the pines, low predawn Psi(leaf), daytime Psi(leaf), and K(l) were associated with low P(n) and G(w). Our data suggest that hydraulic limitations are important in reducing P(n) in old-growth ponderosa pine in northern Arizona, and indicate greater avoidance of soil water stress and greater tolerance of atmospheric water stress by old-growth Gambel oak than by old-growth ponderosa pine. PMID- 12651522 TI - Spatial and temporal variability of foliar mineral concentration in beech (Fagus sylvatica) stands in northeastern France. AB - Foliar mineral concentration may provide a basis for monitoring the consequences of long-term environmental changes, such as eutrophication and acidification of soils, or increase in atmospheric CO(2) concentration. However, analytical drifts and inter-tree and year-to-year variations may confound environmental effects on long-term changes in foliar mineral concentration. We have characterized the relative effects of these potentially confounding factors on foliar carbon, nitrogen, phosphorus, calcium, potassium, magnesium and manganese concentrations in 118 pure beech (Fagus sylvatica L.) stands, sampled in 1969-71 and 1996-97. Interannual fluctuations of these elements were quantified in a subset of six beech stands monitored for 5 years. Intercalibration between the methods used at each sampling period for nitrogen and phosphorus analyses showed significant, but low, relative differences (0.8 and 3.3% for N and P, respectively). Based on inter-tree variability, elements could be arranged in four groups: C (constant), N and P (low variability), K and Ca (medium variability), Mn and Mg (high variability). Inter-tree coefficients of variation were 2, 6, 8, 15, 18, 22 and 27%, respectively. Year-to-year fluctuations increased in the order N, P, Mg, K, Ca, and Mn coefficients of variation of 4, 4, 7, 9, 11, 15 and 29%, respectively). Between the two sampling periods, foliar N concentration increased 12%, whereas decreases were observed for P (-23%), Mg (-38%) and Ca (-16%). Ratios of N/P, N/K and N/Mg increased by 42, 19 and 77%, respectively. These changes were larger than the interannual variations for P, Mg, N/P, N/Mg and Mg/Ca. Decreasing concentrations of P and cations were particularly marked for trees growing on acidic soils, whereas the positive N trend did not depend on soil type. Both increasing atmospheric CO(2) concentrations and acidification of forest soils could contribute to decreasing P and cation concentrations in foliage. The increase in foliar N concentration with time suggests a nitrogen deposition effect. Whatever the causes of these changes, the large shift in element ratios indicates an accelerating imbalance between nitrogen and cation status. PMID- 12651523 TI - Assessing forest soil CO(2) efflux: an in situ comparison of four techniques. AB - A dynamic, closed-chamber infrared gas analysis (IRGA) system (DC-1: CIRAS-1, PP Systems, Hitchin, U.K.) was compared with three other systems for measuring soil CO(2) efflux: the soda lime technique (SL), the eddy correlation technique (EC), and another dynamic, closed-chamber IRGA system (DC-2: LI-6250, Li-Cor, Inc., Lincoln, NE). Among the four systems, the DC-1 systematically gave the highest flux rates. Relative to DC-1, SL, EC and DC-2 underestimated fluxes by 10, 36 and 46%, respectively. These large and systematic differences highlight uncertainties in comparing fluxes from different sites obtained with different techniques. Although the three chamber methods gave different results, the results were well correlated. The SL technique underestimated soil CO(2) fluxes compared with the DC-1 system, but both methods agreed well when the SL data were corrected for the underestimation at higher fluxes, indicating that inter-site comparisons are possible if techniques are properly crosscalibrated. The EC was the only system that was not well correlated with DC-1. Under low light conditions, EC values were similar to DC-1 estimates, but under high light conditions the EC system seriously underestimated soil fluxes. This was probably because of interference by the photosynthetic activity of a moss layer. Although below-canopy EC fluxes are not necessarily well suited for measuring soil CO(2) efflux in natural forest ecosystems, they provide valuable information about understory gas exchange when used in tandem with soil chambers. PMID- 12651524 TI - Effects of nitrogen source and defoliation on growth and biological dinitrogen fixation of Gliricidia sepium seedlings. AB - Effects of four N sources and two defoliation treatments on growth and nitrogenase activity of Gliricidia sepium (Jacq.) Walp seedlings were studied in a greenhouse. All nutrients were supplied in irrigation water to the sterile growing medium. The N sources were: (1) 100 mg l(-1) of N supplied as NO(3) (-) (high-NO(3) (-)), (2) 50 mg l(-1) of N supplied as NO(3) (-) and inoculation with Rhizobium spp. medium-NO(3) (-)), (3)100 mg l(-1) of N supplied as NH(4)NO(3), and (4) inoculation with Rhizobium spp without mineral N (N(2)). At 35 weeks after sowing, mean total biomass was 130.5, 50.5, 22.9 and 17.4 g seedling(-1) in the NH(4)NO(3), N(2), medium-NO(3) (-) and high-NO(3) (-) treatments, respectively. The root/shoot ratio was high in all of the N treatments (1.73 2.77) because the seedlings had big taproots. The medium-NO(3) (-) treatment completely inhibited nodulation, whereas seedlings in the N(2) treatment were profusely nodulated. At 32 weeks after sowing, groups of seedlings in the N(2) and high-NO(3) (-) treatments were subjected to 50 or 100% defoliation. Closed chamber acetylene reduction assays of intact root systems were conducted to compare nitrogenase activity at 7, 14 and 28 days after defoliation (DAD). At 7 and 14 DAD, nitrogenase activity of completely and partially defoliated seedlings was about 10 and 60%, respectively, of that of undefoliated controls. At 28 DAD, nitrogenase activity of completely defoliated seedlings was twice the predefoliation value, whereas nitrogenase activity of partially defoliated seedlings was only 87% of the predefoliation value. Recovery of nitrogenase activity was strongly correlated with foliage regrowth in the completely defoliated seedlings, but not in the partially defoliated seedlings. Abundant belowground C and N reserves in the large taproot probably contributed to the rapid recovery from defoliation. Accumulation of belowground biomass may also improve defoliation tolerance of mature trees. PMID- 12651525 TI - Influence of forage harvesting regimes on dynamics of biological dinitrogen fixation of a tropical woody legume. AB - Effects of three forage harvesting regimes-total removal of foliage and branches once (T-12) or twice a year (T-6) and 50% removal every 2 months (P-2)-on growth and biological dinitrogen fixation of Gliricidia sepium (Jacq.) Walp were studied under subhumid tropical conditions in Guadeloupe, French Antilles. Gliricidia sepium was grown in association with the perennial C(4) grass Dichantium aristatum (Poir) C.E. Hubbard in a two-storied fodder production system. The medium-term effects of pruning on N(2) fixation were assessed by the (15)N natural abundance method. Gmelina arborea Roxb. was used as the non-fixing reference. The trees in the T-12 regime followed the natural phenological cycle, and flowering and podfilling at the beginning of the dry season reduced both foliage and nodule biomass. The T-6 regime impeded flowering, and only a few flowers, on older branches, were produced in the P-2 regime. In trees in the T 12, T-6, and P-2 regimes, fixed N comprised 54-87, 54-92, and 60-87%, respectively, of the total N in aboveground biomass, depending on sampling date. Total annual accumulation of N in harvestable aboveground biomass was highest in trees in the T-6 regime at 313 kg ha(-1), of which 204 kg ha(-1) of N was fixed from the atmosphere. In all treatments, about 70% of the N exported per year from the plot in the fodder harvest came from N(2) fixation. Thus, N(2) fixation makes an important contribution to the N economy of the G. sepium-D. aristatum forage production system, and greatly reduces the need for fertilizer application. PMID- 12651526 TI - Light spectral composition in a tropical forest: measurements and model. AB - I present a simple model that simulates vertical variations in the light spectrum within a forest canopy. The model considers only the vertical, downward transmission of light. The light in each canopy level was assumed to consist of non-intercepted radiation and radiation intercepted within the level and transmitted. The spectrum of non-intercepted light in each canopy level is the same as that of incident light above the canopy (input parameter), whereas the spectrum of transmitted light depends on leaf area index (LAI) and the mean transmission spectrum of leaves. The model was tested in a forest and provided adequate predictions of measured values. Stronger deviations were produced in the near infrared (NIR) waveband in lower canopy levels. Multiple regression between LAI, as the dependent variable, and spectral characteristics (Blue, Green, Red and NIR intensities) had an r(2) of 0.926. As a complement to other methods, I suggest light spectrum analysis as a non-destructive technique for estimating LAI in forest canopies. PMID- 12651527 TI - Influence of temperature and leaf-to-air vapor pressure deficit on net photosynthesis and stomatal conductance in red spruce (Picea rubens). AB - The roles of temperature (T) and leaf-to-air vapor pressure deficit (VPD) in regulating net photosynthesis (A(net)) and stomatal conductance (G(s)) of red spruce (Picea rubens Sarg.) were investigated in a field study and in a controlled environment experiment. Both A(net) and G(s) exhibited a relatively flat response to temperatures between 16 and 32 degrees C. Temperatures between 32 and 36 degrees C markedly decreased both A(net) and G(s). Vapor pressure deficits above 2 kPa had significant effects on both A(net) and G(s). The influence of VPD on A(net) and G(s) fit a linear response model and did not interact significantly with T effects. PMID- 12651528 TI - Radiation-use efficiency and dry matter partitioning of a young olive (Olea europaea) orchard. AB - Radiation-use efficiency (RUE) relates biomass production to the photosynthetically active radiation (PAR) intercepted by a plant or crop. We determined RUE and biomass partitioning coefficients of young olive (Olea europaea) trees for use in a general growth model. In 1995, 1-year-old olive trees var. 'Picual' were planted at a density of either 0.5 or 2.0 trees m(-2) near Cordoba, Spain, at a site providing favorable growth conditions. During the experiment (1995-1997), both PAR interception by the canopy and plant area index (PAI) were measured with radiation sensors. Regular harvests were performed to determine leaf area and biomass accumulation in roots, wood (stem, branches and trunk) and leaves. Leaf, wood and root biomass partitioning coefficients were calculated. The leaf area partitioning coefficients were also estimated. Dry matter production was linearly related to cumulative intercepted PAR. Seasonal RUE, calculated as the slope of the regression of aboveground biomass and cumulative intercepted PAR, was 1.35 g (MJ PAR)(-1). Radiation-use efficiency appeared to respond to environmental conditions, but was independent of planting density and PAI. The young olive trees allocated 0.26 of their total biomass to roots. Partitioning of aboveground dry matter was 0.60 to wood and 0.37 to leaves. As competition increased, dry matter partitioning to wood increased to 0.70. PMID- 12651529 TI - Effects of water status and soil fertility on the C-isotope signature in Pinus radiata. AB - The efficiency with which trees use water is a major determinant of growth under water-limited conditions. We investigated whether increased access to water and nutrients alters water-use efficiency in Pinus radiata D. Don. Intrinsic transpiration efficiency, defined here as the ratio of CO(2) assimilated and water transpired at a given vapor pressure deficit, is determined by the difference between ambient atmospheric CO(2) concentration (c(a)) and leaf intercellular CO(2) concentration (c(i)). The mean value of c(i)/c(a) can be inferred from an analysis of carbon isotope discrimination (Delta) in wood samples. A total of 117 trees, growing at sites with widely varying soil and climatic conditions in Australia and New Zealand, were cored and distinct annual rings were analyzed for their carbon isotope ratio, and correlated with rainfall during the July-June growing season in the year in which the wood was grown. Where possible, carbon isotope ratios were compared for different years within the same trees. The c(i)/c(a) ratio decreased with decreasing water availability, suggesting that intrinsic transpiration efficiency increased with decreasing water availability. An increase in growing season rainfall of 900 mm resulted in an increase in Delta of about 2.0 per thousand, corresponding to a decrease in intrinsic transpiration efficiency of approximately 24%. A stronger relationship was obtained when carbon isotope discrimination was expressed as a function of the ratio of rainfall to potential transpiration. Carbon isotope discrimination was also negatively correlated with mean annual vapor pressure deficit at different sites. In contrast, nutrient availability had no significant effect on carbon isotope discrimination. PMID- 12651530 TI - Transpiration and water relations of poplar trees growing close to the water table. AB - Sap flow was measured on five branches of two poplar (Populus trichocarpa Torr. & A. Gray x P. tacamahaca L.) trees from June to September 1994 in the south of England with stem-surface, heat balance gauges, and was scaled up to estimate transpiration from single trees on the basis of leaf area. On six days, stomatal conductance and plant water potential were measured simultaneously with a porometer and pressure chamber, respectively. The effects of solar radiation (S), vapor pressure deficit (D) and stomatal conductance on transpiration were evaluated. Sap flow per unit leaf area (F(a)) was closely related to the time course of demand attributable to S and D throughout the season, and only slightly affected by the water content of the top 120 cm of soil. Although F(a) increased linearly at low values of D, it showed a plateau with increases in D above 1.2 kPa. The canopy coupling coefficient (1 - Omega) ranged from 0.48 to 0.78 with a mean of 0.65 +/- 0.01, indicating that transpiration was controlled more by stomatal conductance than by incident radiation. The seasonal pattern of tree water loss followed potential evaporation with a peak in late June or early July. On bright days, daily transpiration over the projected crown area was 3.6 mm early in the season, 3.8 mm in mid-season, and 2.7 mm late in the season. The water balance of the system indicated that poplar trees took 15-60% of water transpired from groundwater, with the proportion increasing as the soil in the unsaturated zone dried out. Access to the water table resulted in high predawn water potentials throughout the season. Estimated hydraulic resistance to water flow in the soil-tree system was in the range of 1.5 to 1.93 x 10(6) MPa s m(-3). PMID- 12651531 TI - Diurnal changes in water conduction in loblolly pine (Pinus taeda) and Virginia pine (P. virginiana) during soil dehydration. AB - We studied diurnal changes in water conduction during soil dehydration in 37 month-old seedlings of one Virginia pine (Pinus virginiana Mill.) and two loblolly pine (P. taeda L.) sources, one from North Carolina (NC) and the other from the "Lost Pines" areas of Texas (TX), in an environmentally controlled growth chamber. For seedlings of similar biomass, the TX source had higher values of transpiration, needle conductance, and plant hydraulic conductivity under well watered conditions than the NC source. Under dry soil conditions, the TX source had lower values of water conduction than the NC source. The Virginia pine source responded similarly to the TX source under both well-watered and dry soil conditions. For all three pine sources, gradients between soil and needle water potentials were greatest when the seedlings were moderately stressed. The TX and Virginia pine sources had higher gradients and lower daytime needle water potentials under moderate stress conditions than the NC source. Predawn needle water potentials did not differ among the pine sources. We conclude that the TX and Virginia pine sources have decreased daytime needle water potentials and increased water potential gradients during the daytime under moderate stress conditions, but with no disruption of recovery at predawn. The greater rates of transpiration and water conduction by the TX source compared with the NC source under well-watered conditions suggest a means by which growth can be maximized prior to the onset of drought, thereby enhancing survival of loblolly pines in drought-prone environments. PMID- 12651532 TI - Genetic variation for carbon isotope composition in honey mesquite (Prosopis glandulosa). AB - Carbon isotope composition (delta(13)C) is a useful surrogate for integrated, plant water-use efficiency (WUE) when measured on plants grown in a common environment. In a variety of species, genetic variation in delta(13)C has been linked to the distribution of genotypes across gradients in atmospheric and soil water. We examined genetic variation for delta(13)C in seedlings of honey mesquite (Prosopis glandulosa Torr.), an invasive grassland shrub that thrives in the southwestern USA. Fifteen maternal families, representing progeny of 15 adult trees, were studied in three common garden experiments in a greenhouse. The 990 km east-west transect along which the adult trees were located encompasses a wide precipitation gradient, and includes mesic grassland, semiarid grassland, and Chihuahuan desert ecosystems. Genetic variation for delta(13)C in mesquite was substantial, with the rank order of half-sib families based on delta(13)C relatively stable across experiments, which were conducted under different environmental conditions. Conversely, rankings of families by mean seedling height (an index of growth rate) varied markedly among experiments. Seedlings derived from Chihuahuan desert adults emerged more quickly and had more negative delta(13)C (indicative of lower WUE) than seedlings derived from the other regions. Although delta(13)C and seedling height were not correlated, these results suggest that mesquite genotypes at the drier, western extreme of the species' range are adapted for quicker emergence and possibly faster growth than genotypes from mesic areas. Together, these traits may facilitate exploitation of infrequent precipitation events. PMID- 12651533 TI - Transpiration increases during the dry season: patterns of tree water use in eucalypt open-forests of northern Australia. AB - Australian savannas exhibit marked seasonality in precipitation, with more than 90% of the annual total falling between October and May. The dry season is characterized by declining soil water availability and high vapor pressure deficits (up to 2.5 kPa). We used heat pulse technology to measure whole-tree transpiration rates on a daily and seasonal basis for the two dominant eucalypts at a site near Darwin, Australia. Contrary to expectations, transpiration rates were higher during the dry season than during the wet season, largely because of increased evaporative demand and the exploitation of groundwater reserves by the trees. Transpiration rates exhibited a marked hysteresis in relation to vapor pressure deficit, which was more marked in the dry season than in the wet season. This result may be attributable to low soil hydraulic conductivity, or the use of stored stem water, or both. Tree water use was strongly correlated with leaf area and diameter at breast height and there were no differences in transpiration between the species studied. These results are discussed in relation to scaling tree water use to stand water use. PMID- 12651534 TI - Direct inhibition of maintenance respiration in western hemlock roots exposed to ambient soil carbon dioxide concentrations. AB - Root respiration often exhibits a direct and immediate decline with increasing concentrations of ambient soil carbon dioxide concentration ([CO(2)]), and recent evidence suggests this decline may be attributable to a decline in maintenance respiration within the root. If true, this concept could provide a clue to the biochemical process underlying respiratory inhibition as well as improve our knowledge of the timing and degree to which this inhibition occurs in nature. To test the hypothesis that maintenance respiration exhibits a direct, negative response to increasing [CO(2)], we measured total respiration in intact root systems of western hemlock (Tsuga heterophylla (Raf.) Sarg.) seedlings grown at different relative growth rates and exposed to soil [CO(2)]s ranging from 91 to 7008 &mgr;mol mol(-1). Analysis of covariance was used to separate maintenance from total respiration. Total respiration declined exponentially with increasing [CO(2)]. Maintenance respiration, which comprised 85% of total respiration over all treatments, also declined exponentially with increasing [CO(2)]. Growth respiration was not inhibited at any [CO(2)]. These findings may explain why roots of some fast-growing species do not show [CO(2)] inhibition. PMID- 12651535 TI - Quenching analysis in poplar clones exposed to ozone. AB - Effects of ozone (O(3)) on photo-inhibition of photosynthesis were investigated in two poplar clones (Populus deltoides Bartr. ex Marsh. x maximowiczii A. Henry clone Eridano, O(3)-sensitive, and P. x euramericana (Dode) Guinier clone I-214, O(3)-resistant) by using pulse amplitude modulation chlorophyll fluorescence. After a 15-day exposure to O(3) at 60 nl l(-1) for 5 h day(-1), the effective photosynthetic quantum yield of photosystem II (PSII), as indicated by the ratio of variable to maximum chlorophyll fluorescence, significantly declined in both young and mature leaves of the two clones. Compared with control samples, mature leaves of both clones showed differences in fluorescence quenching and quantum yield of PSII when exposed to ozone fumigation. We conclude that photo-inhibition is caused by transformation of active reaction centers to photochemically inactive centers that dissipate excitation energy into heat, thus causing non photochemical fluorescence quenching. The changes observed in chlorophyll a fluorescence of O(3)-treated young leaves in both clones are consistent with O(3) having a negligible effect on photosynthetic performance, but inducing a premature decrease in chlorophyll a concentration. A possible role of O(3) pollution on chlorophyll fluorescence yield is discussed. PMID- 12651536 TI - Statistical evaluation of bud development theories: application to bud burst of Betula pendula leaves. AB - Resampling methods were used to evaluate models based on alternative bud development theories applied to Betula pendula Roth data. Statistical testing based on the bootstrap method showed that the mean square errors (MSE) of the predicted bud-burst dates of two models, in which the start of ontogenetic development depended on dormancy development only, did not differ significantly. However, the MSE of the model in which the start of ontogenesis depended on a signal from light climate, indicated by using a fixed calendar date, was significantly smaller than that of the models depending on dormancy development. Model parameters were highly multi-collinear; i.e., sensitive to changes in the data. The cross-validation method was used to determine the prediction error of the models. The predictive ability of the models was not much less for an independent data set than for the original data. PMID- 12651537 TI - Can differences in root responses to soil drying and compaction explain differences in performance of trees growing on landfill sites? AB - Two tropical woody species, Acacia confusa Merrill and Litsea glutinosa (Lour.) C.B. Robinson, were grown under controlled conditions in PVC pipes filled with John Innes No. 2 soil. To investigate root distribution, physiological characteristics and hydraulic conductivity, four soil treatments were imposed well-watered and noncompacted (control), well-watered and compacted; unwatered and noncompacted, and unwatered and compacted. In L. glutinosa, rooting depth and root elongation were severely restricted when soil bulk density increased from around 1.12 to 1.62 g cm(-3), whereas soil compaction had little effect on these parameters in A. confusa. As soil drying progressed, root water potential and osmotic potential declined more slowly in L. glutinosa than in A. confusa. Both the soil drying and compaction treatments significantly stimulated the accumulation of root abscisic acid (ABA) in both species. Soil drying damaged the root cell membrane of A. confusa, but had little influence on the root cell membrane of L. glutinosa. Soil drying had a greater effect on root hydraulic conductivity (L(p)) in L. glutinosa than in A. confusa, whereas the effect of soil compaction on L(p) was less in L. glutinosa than in A. confusa. Soil drying enhanced the effects of soil compaction on root L(p). We conclude that soil drying and compaction have large species-specific effects on the distribution, growth and physiology of roots. The relationships of these root properties to the species' ability to tolerate unfavorable soil conditions were examined. PMID- 12651538 TI - Parameterization and testing of a biochemically based photosynthesis model for walnut (Juglans regia) trees and seedlings. AB - The biochemically based leaf photosynthesis model proposed by Farquhar et al. (1980) and the stomatal conductance model proposed by Jarvis (1976) were parameterized for walnut. Responses of photosynthesis to CO(2) and irradiance were used to determine the key parameters of the photosynthesis model. Concurrently, stomatal conductance responses to leaf irradiance (Q), leaf temperature (T(l)), water vapor pressure deficit at the leaf surface (D), and air CO(2) concentration at the leaf surface (C(s)) were used to parameterize the stomatal conductance model. To test the generality of the model parameters, measurements were made on leaves from a 20-year-old tree growing in the field, and from sunlit and shaded greenhouse-grown seedlings. The three key parameters of the photosynthesis model (maximum carboxylation rate V(cmax), electron transport capacity J(max), and dark respiration rate R(d)) and the key parameter of the conductance model (reference stomatal conductance, g(sref)) were linearly correlated with the amount of leaf nitrogen per unit leaf area. Unique relationships could be used to describe nitrogen effects on these parameters for leaves from both the tree and the seedlings. Our data allowed separation of the effects of increasing total photosynthetic apparatus per unit leaf area from the effects of partitioning nitrogen among different pools of this apparatus for foliage acclimation to leaf irradiance. Strong correlations were found between stomatal conductance g(s) and Q, D and C(s), whereas the relationship between g(s) and T(l) was weak. Based on these parameterizations, the model adequately predicted leaf photosynthesis and stomatal conductance when tested with an independent set of data obtained for the tree and seedlings. Total light-driven electron flows derived from chlorophyll fluorescence data obtained at different leaf temperatures were consistent with values computed by the model. The model was also tested with branch bag data acquired from a three-year-old potted walnut tree. Despite a relatively large variance between observed and simulated values, the model predicted stomatal conductance and photosynthesis reasonably well at the branch scale. The results indicate that the photosynthesis-conductance model developed here is robust and can be applied to walnut trees and seedlings under various environmental conditions where water is non-limiting. PMID- 12651539 TI - Transpiration by two poplar varieties grown as coppice for biomass production. AB - Fast-growing tree clones selected for biomass plantations are highly productive and therefore likely to use more water than the agricultural crops they replace. We report field measurements of transpiration through the summer of 1994 from two poplar clones, Beaupre (Populus trichocarpa Torr. & A. Gray x P. deltoides Bartr. ex Marsh.) and Dorschkamp (P. deltoides x P. nigra L.), grown as unirrigated short-rotation coppice in southern England. Stand transpiration was quantified by scaling up from sap flow measurements made with the heat balance method in a sample of stems. Leaf conductances, leaf area development, meteorological variables and soil water deficit were also measured to investigate the response of the trees to the environment. High rates of transpiration were found for Beaupre. In June, when soil water was plentiful, the mean (+/- SD) transpiration rate over an 18-day period was 5.0 +/- 1.8 mm day(-1), reaching a maximum of 7.9 mm day(-1). Transpiration rates from Dorschkamp were lower, as a result of its lower leaf area index. High total leaf conductances were measured for both Beaupre (0.34 +/- 0.17 mol m(-2) s(-1)) and Dorschkamp (0.39 +/- 0.16 mol m(-2) s(-1)). Leaf conductance declined slightly with increasing atmospheric vapor pressure deficit in both clones, but only in Beaupre did leaf conductance decrease as soil water deficit increased. PMID- 12651540 TI - Stomatal behavior of four woody species in relation to leaf-specific hydraulic conductance and threshold water potential. AB - Midday stomatal closure is mediated by the availability of water in the soil, leaf and atmosphere, but the response to these environmental and internal variables is highly species specific. We tested the hypothesis that species differences in stomatal response to humidity and soil water availability can be explained by two parameters: leaf-specific hydraulic conductance (K(L)) and a threshold leaf water potential (Psi(threshold)). We used a combination of original and published data to estimate characteristic values of K(L) and Psi(threshold) for four common tree species that have distinctly different stomatal behaviors: black cottonwood (Populus trichocarpa Torr. & Gray.), Douglas fir (Pseudotsuga menziesii (Mirb.) Franco), red alder (Alnus rubra Bong.) and western hemlock (Tsuga heterophylla (Raf.) Sarg.). We used the values to parameterize a simple, nonelastic model that predicts stomatal conductance by linking hydraulic flux to transpirational flux and maintaining Psi(leaf) above Psi(threshold). The model successfully predicted fundamental features of stomatal behavior that have been reported in the literature for these species. We conclude that much of the variation among the species in stomatal response to soil and atmospheric water deficits can be explained by K(L) and Psi(threshold). The relationship between Psi(threshold) and xylem vulnerability to cavitation differed among these species. PMID- 12651541 TI - Analysis of gas exchange of Merkus pine populations by the optimality approach. AB - We hypothesized that northern and northeastern Thai populations of Merkus pine (Pinus merkusii Jungh. et de Vriese), which differ in the duration of their grass stage and which originate from slightly different climates in terms of water availability, differ in their gas exchange characteristics. We compared CO(2) exchange response to irradiance, diurnal regulation of leaf conductance within a 10-day period and structural properties among the populations. We used a model, which is based on the concept of optimal stomatal regulation, to analyze CO(2) exchange and transpiration rates. The two geographical groups did not differ in CO(2) exchange response to irradiance, diurnal transpiration, or water use. Mean stomatal length was significantly greater in the northeastern population than in the northern populations, but stomatal frequency did not differ among populations. First-year shoot growth and dry matter production, allocation of nitrogen to needles and root:shoot ratios were similar in the two geographical groups. Genotypic variation in the duration of the grass stage was not reflected in variation in gas exchange, indicating that the grass stage is an adaptation to more site-specific conditions. The modeled response of CO(2) exchange rates to irradiance fitted well the rates measured under laboratory conditions. The transpiration model, which utilized maximum leaf conductance and other parameters derived from the CO(2) exchange measurements, also fitted well the transpiration rates measured in a greenhouse under changing environmental conditions. PMID- 12651542 TI - Acclimation of loblolly pine (Pinus taeda) seedlings to high temperatures. AB - To determine the extent to which loblolly pine seedlings (Pinus taeda L.) acclimate to high temperatures, seedlings from three provenances-southeastern Texas (mean annual temperature 20.3 degrees C), southwestern Arkansas (mean annual temperature 16.2 degrees C) and Chesapeake, Maryland (mean annual temperature 12.8 degrees C)-were grown at constant temperatures of 25, 30, 35 or 40 degrees C in growth chambers. After two months, only 14% of the seedlings in the 40 degrees C treatment survived, so the treatment was dropped from the experiment. Provenance and family differences were not significant for most measured variables. Total biomass was similar in the 25 and 30 degrees C treatments, and less in the 35 degrees C treatment. Foliage biomass was higher, and root biomass lower, in the 30 degrees C treatment compared with the 25 degrees C treatment. Net photosynthesis and dark respiration of all seedlings were measured at 25, 30 and 35 degrees C. Both net photosynthesis and dark respiration exhibited acclimation to the temperature at which the seedlings were grown. For each temperature treatment, the highest rate of net photosynthesis was measured at the growth temperature. Dark respiration rates increased with increasing measurement temperature, but the basal rate of respiration, measured at 25 degrees C, decreased from 0.617 &mgr;mol m(-2) s(-1) in the 25 degrees C treatment to 0.348 &mgr;mol m(-2) s(-1) in the 35 degrees C treatment, resulting in less carbon loss in the higher temperature treatments than if the seedlings had not acclimated to the growth conditions. Temperature acclimation, particularly of dark respiration, may explain why total biomass of seedlings grown at 30 degrees C was similar to that of seedlings grown at 25 degrees C. PMID- 12651543 TI - Influence of ozone on cold acclimation in sugar maple seedlings. AB - During summer 1994, sugar maple (Acer saccharum Marsh.) seedlings were grown in open-top chambers supplied with air containing near ambient ozone concentration (control, low O(3)) or three times the ambient ozone concentration (high O(3)). The rate of CO(2) assimilation was significantly reduced by chronic exposure to a high concentration of ozone during the summer. During fall, seedlings were removed from the open-top chambers and acclimated to cold under natural conditions. In both species during cold acclimation, the starch concentration decreased, whereas the sucrose concentration increased. There was no treatment effect on the freezing tolerance of roots, even though roots in the high-O(3) treatment accumulated higher concentrations of the cryoprotective oligosaccharides raffinose and stachyose than control roots. Cold acclimation occurred earlier and stachyose concentration of stems was higher in high-O(3) treated seedlings than in low-O(3)-treated seedlings. Cold acclimation was associated with an earlier accumulation of ABA in the xylem sap of high-O(3) treated seedlings compared with low-O(3)-treated seedlings. PMID- 12651544 TI - Magnesium nutrition and photosynthesis in Pinus radiata: clonal variation and influence of potassium. AB - Magnesium (Mg) nutrition and photosynthesis were studied in clones of Pinus radiata D. Don grown in sand culture for 21 weeks at four Mg concentrations (0.008, 0.04, 0.2 and 0.4 mM) and three potassium (K) concentrations (0.25, 0.5 and 2.5 mM). We found significant clonal variation in Mg nutrition of P. radiata. Plants grown at 0.04 mM [Mg] or less showed pronounced visible symptoms of foliar Mg deficiency. Net photosynthetic rate and leaf conductance were closely related to shoot Mg concentrations below a concentration of 0.6 mg Mg g(DW) (-1). Potassium enhanced the development of visible symptoms of foliar Mg deficiency. At the lowest Mg concentration tested (0.008 mM), the severity of needle chlorosis and necrosis increased with increasing K concentration in the culture solution. With increasing Mg concentration, 2.5 mM [K] in the culture solution markedly increased root Mg concentration, but decreased shoot Mg concentration, suggesting that excessive K inhibited Mg mobilization from roots to shoots. Rates of growth and photosynthesis were both severely inhibited at 0.008 mM [Mg]. PMID- 12651545 TI - Green fluorescent protein as a tool for monitoring transgene expression in forest tree species. AB - The gene coding for green fluorescent protein (GFP) from the jellyfish Aequorea victoria was successfully used as a vital marker for the transformation of three woody plant species, black spruce (Picea mariana (Mill.) BSP), white pine (Pinus strobus L.) and poplar (Populus spp.). The gfp gene and the gene conferring resistance to kanamycin (nptII) were introduced by microprojectile bombardment or Agrobacterium tumefaciens-mediated technology. Screening by fluorescence microscopy of the transformed plant material, under the selection of kanamycin, identified five to eight cell lines from each tree species that clearly expressed GFP. Expression of GFP was observed in somatic embryonal cells of the coniferous species and in stem sections of poplar. For all species, GFP transgene expression was stable over multiple subcultures. Stable integration of the gfp gene into plant genomes was confirmed by Southern hybridization or polymerase chain reaction (PCR) analysis. We conclude that GFP can be used as a vital marker and reporter protein in transformation experiments with gymnosperms and deciduous trees. PMID- 12651546 TI - A micro-power precision amplifier for converting the output of light sensors to a voltage readable by miniature data loggers. AB - To record photosynthetically active radiation (PAR) simultaneously at a number of points throughout a forest canopy, we developed a simple, inexpensive (< $10 US) current-to-voltage converter that processes the current generated by a photodiode radiation sensor to a voltage range that is recordable with a miniature data logger. The converter, which weighs less than 75 g and has a volume of only 100 cm(3), is built around an ultra-low power OP-90 precision operational amplifier, which consumes less than 0.5 mA at 9 V when converting the output of a Li-Cor LI 190SA quantum sensor exposed to photosynthetically active radiation (PAR) of 2500 &mgr;mol m(-2) s(-1) or only 5 &mgr;A in low light. A small 9-V battery thus powers the amplifier for more than 1000 h of continuous operation. Correlations between photometer readings and voltage output from the current-to-voltage converter were high and linear at both high and low PAR. Sixteen Li-Cor LI-190SA quantum sensors each equipped with current-to-voltage converters and connected to a miniature data logger were deployed in the upper branches of a Panamanian tropical rainforest canopy. Each unit performed reliably during a one- or two week evaluation. PMID- 12651547 TI - Canopy photosynthesis of sugar maple (Acer saccharum): comparing big-leaf and multilayer extrapolations of leaf-level measurements. AB - A comparison is made between a big-leaf model (i.e., without details of the canopy profile) and two multilayer models (i.e., with details of the canopy profile) to estimate daily canopy photosynthesis of a sugar maple (Acer saccharum Marsh.) stand. The first multilayer model uses the distribution of leaf area by leaf mass per unit area (LMA) classes, the observed relationships between the parameters of a photosynthesis-irradiance curve and LMA, and the relationship between relative irradiance and LMA to estimate canopy photosynthesis. When compared with this model, the big-leaf model underestimates daily canopy photosynthesis by 26% because of an assumed proportionality between photosynthetic capacity and relative irradiance, a proportionality that is inconsistent with our data. The bias induced by this assumption is reduced when the big-leaf model is compared with the second multilayer model which, in addition to the assumptions made for the first multilayer model, accounts for the sunlit and shaded fractions of leaf area. The residual bias is almost eliminated when the big-leaf model is run using a weekly averaged irradiance. It is likely, however, that this is the result of a compensating bias that, in this particular case, compensates for the initial bias introduced by the proportionality assumption. It is also shown that canopy photosynthesis can be represented by spatially inexplicit multilayer models that use leaf mass per area as a covariable to describe leaf characteristics and environment. Such models represent an interesting alternative to the biased big-leaf approach. PMID- 12651548 TI - Gas exchange and carbon isotope discrimination of Juniperus osteosperma and Juniperus occidentalis across environmental gradients in the Great Basin of western North America. AB - We determined how ecophysiological characteristics of two juniper species, Juniperus occidentalis Hook. (western juniper) and Juniperus osteosperma (Torr.) Little (Utah juniper), changed along altitudinal and regional environmental gradients in the Great Basin of western North America. We obtained diurnal measurements of leaf gas exchange and xylem water potential (Psi) from plants at a low and a high altitude site within each of six mountain ranges during fall 1994, spring, summer, and fall 1995, and summer 1996. We also determined carbon isotope composition (delta(13)C) of leaf cellulose produced during the 1995 growing season. Overall, leaf gas exchange, Psi and delta(13)C did not differ significantly between species. Differences in daily (A(d)) and season-long (A(s)) carbon assimilation among mountain ranges suggested two groupings-a group of northern ranges and a group of southern ranges. Each group contained one mountain range with J. occidentalis and two with J. osteosperma. Differences in carbon assimilation based on this grouping were associated with two findings: (1) conductance of CO(2) from substomatal cavities to the site of carboxylation (g(m)) for junipers in the northern ranges averaged almost twice that of junipers in the southern ranges; and (2) physiological shifts occurred such that A(d) of junipers in the northern ranges was influenced more by Psi(pd), whereas A(d) of junipers in the southern ranges was influenced more by leaf temperature. Mean delta(13)C over all trees at a site was significantly correlated with annual precipitation. Significant differences in A(d) occurred between altitudes, but these differences were associated with differences in the timing of optimum leaf temperature for photosynthesis rather than with physiological acclimation to temperature, irradiance, or Psi. Most gas exchange parameters (e.g., assimilation, transpiration, stomatal conductance, and water use efficiency) varied seasonally, and the seasonal differences were strongly influenced by water stress. PMID- 12651549 TI - Boundary layer conductance, leaf temperature and transpiration of Abies amabilis branches. AB - We used three methods to measure boundary layer conductance to heat transfer (g(bH)) and water vapor transfer (g(bV)) in foliated branches of Abies amabilis Dougl. ex J. Forbes, a subalpine forest tree that produces clumped shoot morphology on sun-formed branches. Boundary layer conductances estimated in the field from energy balance measurements increased linearly from approximately 10 mm s(-1) at low wind speeds (< 0.1 m s(-1)) to over 150 mm s(-1) at wind speeds of 2.0 m s(-1). Boundary layer conductances measured on shoot models in a wind tunnel were consistently higher than field measurements. The difference between wind tunnel values and field measurements was attributable to variation in path length between the two experimental environments. Boundary layer conductance estimated by subtracting stomatal resistance (r(sV)) measured with a porometer from the total branch vapor phase resistance were unusually small. Sensitivity analysis demonstrated that this method is not suitable for coniferous foliage or when stomatal conductance (g(sV)) is small compared with g(bV). Analysis of the relative magnitudes of g(sV) and g(bV) revealed that, under most conditions, A. amabilis branches are well coupled (i.e., g(sV) is the dominant controller of transpiration). The boundary layer conductance to heat transfer is small enough that leaf temperature can become substantially higher than air temperature when radiation is high and wind speed is low. Over a two-month period, the maximum difference between leaf and air temperatures exceeded 6 degrees C. Leaf temperature exceeded air temperature by more than 2 degrees C on 10% of the daylight hours during this period. Consideration of both the photosynthetic temperature response of A. amabilis foliage as well as the summer air temperature conditions in its habitat suggests that these elevated leaf temperatures do not have a significant impact on carbon gain during the growing season. PMID- 12651550 TI - Hydraulic conductances of angiosperms versus conifers: similar transport sufficiency at the whole-plant level. AB - Bond's "slow seedling" hypothesis proposes that, because of slow growth rates caused by an inefficient transport system and low leaf photosynthetic capacity, gymnosperm seedlings are weak competitors with angiosperms in productive habitats. We measured component (shoot, leaf, and root) and whole-plant hydraulic conductances of sapling-sized tropical plants growing on nitrogen-poor white sand in Borneo. After accounting for size effects, there were no significant differences in conductances between evergreen angiosperms (nine species) and conifers (three species). Plant successional status or transpiration rate seemed more important than soil fertility in determining hydraulic conductance colonizers had significantly higher whole-plant conductance than late-succession species. Contrary to prediction, leaf hydraulic conductance (normalized by projected leaf area) was unrelated to complexity of venation in conifers and angiosperms, but was highly correlated with whole-plant conductance. Analyses of published data showed that leafless branches of temperate deciduous angiosperms had higher leaf-area normalized hydraulic conductivity than conifers, but there was no significant difference in adult, whole-plant conductance between these taxa. Thus, at the branch level, conifers with narrow tracheids have less efficient transport than angiosperms with wider vessels, but variations in other resistance components and hydraulic architecture (e.g., sapwood/leaf area ratio) ultimately equalize the sufficiency of water transport to leaves of conifers and angiosperms. Although failing to support one of the proposed mechanisms, our findings did not refute the "slow seedling" hypothesis per se. PMID- 12651551 TI - Regulation of water loss in populations of Populus trichocarpa: the role of stomatal control in preventing xylem cavitation. AB - Variations in resistance to drought-induced xylem cavitation, xylem air-entry points, stomatal behavior, and hydraulic conductivity were measured in four populations of Populus trichocarpa Torr. & A. Gray collected along an east-west humidity and temperature gradient in Washington State, USA. Xylem air-entry points were less negative in trees from moist environments (-0.71 and -1.32 MPa in the Hoh and Nisqually populations, respectively) than in trees from dry environments (-1.55 and -1.67 MPa in the Palouse and Yakima populations, respectively). Xylem cavitation in response to experimental drought was consistent with air-injection measures of xylem air-entry points for a given population. Populations vulnerable to cavitation also exhibited higher stem specific hydraulic conductivities and limited stomatal control compared with resistant populations. Populations exhibiting vulnerability to cavitation and limited stomatal control desiccated more rapidly during drought compared with resistant populations. This study provides evidence of interpopulation variation in resistance to drought-induced xylem cavitation, stomatal behavior, and hydraulic conductivity within Populus trichocarpa. PMID- 12651552 TI - Polyphenols as potential markers to differentiate juvenile and mature chestnut shoot cultures. AB - The phenolic contents of eight in-vitro-cultivated chestnut clones (Castanea sativa Mill. and C. sativa x C. crenata Siebold & Zucc. hybrids) were analyzed both qualitatively and quantitatively. The aim of the work was to identify potential phenolic markers of: (i) juvenile or mature state; (ii) topophysical origin; and (iii) rooting capacity. A condensed tannin was detected in mature material but not in juvenile material, indicating that it could be used as a qualitative marker. Other qualitative phenolic differences were found between basal shoots and crown shoots of some clones, but it was not possible to discriminate among these materials in a general way. Canonical discriminant analysis was used for the study of quantitative markers. Differentiation between mature and juvenile material, and between materials differing in in vitro rooting capacity was possible according to the results of the analysis. Nevertheless, no significant quantitative differences were found between the phenolic content of material of basal shoot origin and that of crown shoot origin, indicating that the greater juvenility of material of basal origin compared with that of crown origin was not reflected in differences in phenolic content. PMID- 12651553 TI - Limitation of leaf carbon gain by stomatal and photochemical processes in the top canopy of Macaranga conifera, a tropical pioneer tree. AB - Diurnal changes in gas exchange and chlorophyll fluorescence were measured in the top canopy leaves of the tropical rainforest tree species, Macaranga conifera (Zoll.) Muell. Arg. during a drought year. Maximum values of net photosynthetic rate (P(n), 10 &mgr;mol m(-2) s(-1)) and stomatal conductance (g(s), 0.2 mol m( 2) s(-1)) were found in east-facing leaves in early morning. After 1000 h, both P(n) and g(s) decreased. Minimum daytime values of P(n), g(s), and photosystem II (PSII) quantum yield (DeltaF/F(m)') were found in horizontally fixed leaves. At a given electron transport rate through PSII (ETR), P(n) was higher in early morning than at midday, suggesting a high rate of photorespiration at midday. We tested the hypothesis that the effect of low leaf temperature (T(leaf)) on P(n) is significant in the early morning, whereas the effect of low g(s) on P(n) predominates at midday. In the early morning, when T(leaf) was increased from 32 to 38 degrees C by artificial heating, P(n) at a given ETR decreased 29%, suggesting that the low T(leaf) was associated with a high P(n). When T(leaf) at midday was decreased from 37 to 32 degrees C by artificial cooling, P(n) increased 22%, but P(n) at a given ETR was higher in early morning than at midday, even at the same low T(leaf) (32 degrees C). This suggests that the rate of photorespiration was higher at midday than in early morning because low g(s) at midday caused a reduction in leaf intercellular CO(2) concentration. We conclude that low P(n) at midday was the result of both a reduction in the photochemical process and an increase in stomatal limitation. PMID- 12651554 TI - Fine root respiration in mature eastern white pine (Pinus strobus) in situ: the importance of CO(2) in controlled environments. AB - We measured seasonal fine root respiration rate in situ while controlling chamber temperature and [CO(2)]. Atmospheric [CO(2)] ([CO(2)](a)) and measured soil [CO(2)] ([CO(2)](s)) were alternately delivered to a cuvette containing intact fine roots of eastern white pine (Pinus strobus L.). Respiration rates were consistently higher in [CO(2)](a) than in [CO(2)](s) and were almost three times higher during midsummer. Respiration rates were immediately reversed after returning to the alternate [CO(2)] (i.e., [CO(2)](a) --> [CO(2)](s) --> [CO(2)](a), and vice versa) suggesting a direct effect of elevated [CO(2)] on apparent respiration. Soil-[CO(2)]-based respiration rates decreased with increasing [CO(2)] on a dry mass and tissue [N] basis. We conclude that estimates of soil CO(2) flux and soil carbon budgets may be improved by more completely accounting for the rhizosphere microclimate (i.e., soil temperature and [CO(2)](s)) during measurement of fine root respiration. PMID- 12651555 TI - Sap-flux-scaled transpiration responses to light, vapor pressure deficit, and leaf area reduction in a flooded Taxodium distichum forest. AB - We used 20-mm-long, Granier-type sensors to quantify the effects of tree size, azimuth and radial position in the xylem on the spatial variability in xylem sap flux in 64-year-old trees of Taxodium distichum L. Rich. growing in a flooded forest. This information was used to scale flux to the stand level to investigate variations in half-hourly and daily (24-hour) sums of sap flow, transpiration per unit of leaf area, and stand transpiration in relation to vapor pressure deficit (D) and photosynthetically active radiation (Q(o)). Measurements of xylem sap flux density (J(s)) indicated that: (1) J(s) in small diameter trees was 0.70 of that in medium and large diameter trees, but the relationship between stem diameter as a continuous variable and J(s) was not significant; (2) J(s) at 20-40 mm depth in the xylem was 0.40 of that at 0-20 mm depth; and (3) J(s) on the north side of trees was 0.64 of that in directions 120 degrees from the north. Daily transpiration was linearly related to daily daytime mean D, and reached a modest value of 1.3 mm day(-1), reflecting the low leaf area index (LAI = 2.2) of the stand. Because there was no soil water limitation, half-hourly water uptake was nearly linearly related to D at D < 0.6 kPa during both night and day, increasing to saturation during daytime at higher values of D. The positive effect of Q(o) on J(s) was significant, but relatively minor. Thus, a second order polynomial with D explained 94% of the variation in J(s) and transpiration. An approximately 40% reduction in LAI by a hurricane resulted in decreases of about 18% in J(s) and stand transpiration, indicating partial stomatal compensation. PMID- 12651556 TI - Biomass investment in leaf lamina versus lamina support in relation to growth irradiance and leaf size in temperate deciduous trees. AB - Foliar biomass investment in support and assimilative compartments was studied in four temperate deciduous tree species along a natural light gradient across the canopy. The species ranked according to shade tolerance as Betula pendula Roth. < Populus tremula L. < Fraxinus excelsior L. < Tilia cordata Mill. Long-term light conditions at sampling locations were characterized as seasonal mean integrated quantum flux density (Q(int), mol m(-2) day(-1)) estimated by a method combining hemispherical photography and light measurements with quantum sensors. Leaf morphology was altered by Q(int) in all species. Both lamina and petiole dry mass per lamina area (LMA and PMA, respectively) increased with increasing Q(int). Shade-tolerant species had lower LMA at low Q(int) than shade-intolerant species; however, PMA was not related to shade tolerance. Across species, the ratio of petiole dry mass to lamina dry mass (PMR) varied from 0.07 to 0.21. It was independent of Q(int) in the simple-leaved species, but decreased with increasing Q(int) in the compound-leaved F. excelsior, which also had the largest foliar biomass investment in petioles. Differences in leaf mass and area, ranging over four orders of magnitude, provided an explanation for the interspecific variability in PMR. Species with large leaves also had greater biomass investments in foliar support than species with smaller leaves. This relationship was similar for both simple- and compound-leaved species. There was a negative relationship between PMR and petiole N concentration, suggesting that petioles had greater carbon assimilation rates and paid back a larger fraction of their construction cost in species with low PMR than in species with high PMR. This was probably the result of a negative relationship between PMR and petiole surface to volume ratio. Nevertheless, petioles had lower concentrations of mineral nutrients than laminas. Across species, the ratio of petiole N to lamina N varied from only 3 to 6%, demonstrating that petiole costs are less in terms of nutrients than in terms of total biomass, and that the petiole contribution to carbon assimilation is disproportionately lower than that of the lamina contribution. PMID- 12651557 TI - Growth, water relations, and survival of drought-exposed seedlings from six maternal families of honey mesquite (Prosopis glandulosa): responses to CO(2) enrichment. AB - Low water availability is a leading contributor to mortality of woody seedlings on grasslands, including those of the invasive shrub Prosopis. Increasing atmospheric CO(2) concentration could favor some genotypes of this species over others if there exists intraspecific variation in the responsiveness of survivorship to CO(2). To investigate such variation, we studied effects of CO(2) enrichment on seedling survival in response to uniform rates of soil water depletion in six maternal families of honey mesquite (P. glandulosa Torr. var. glandulosa). Three families each from the arid and mesic extremes of the species' distribution in the southwestern United States were studied in environmentally controlled glasshouses. Relative water content at turgor loss and osmotic potential were not affected by CO(2) treatment. Increased atmospheric CO(2) concentration, however, increased growth, leaf production and area, and midday xylem pressure potential, and apparently reduced transpiration per unit leaf area of seedlings as soil dried. Consequently, CO(2) enrichment about doubled the fraction of seedlings that survived soil water depletion. Maternal families of honey mesquite differed in percentage survival of drought and in several other characteristics, but differences were of similar or of smaller magnitude compared with differences between CO(2) treatments. There was no evidence for genetic variation in the responsiveness of survivorship to CO(2). By increasing seedling survival of drought, increasing atmospheric CO(2) concentration could increase the abundance of honey mesquite where establishment is limited by water availability. Genetic types with superior ability to survive drought today, however, apparently will maintain that advantage in the future. PMID- 12651558 TI - Shoot water relations of mature black spruce families displaying a genotype x environment interaction in growth rate. I. Family and site effects over three growing seasons. AB - Pressure-volume curves were determined for black spruce (Picea mariana (Mill.) BSP) trees from four full-sib families. During the first two years, trees were measured from a plantation on a dry site. In the third year, trees were sampled from the dry site and a wet site. Diurnal measurements of shoot water potential allowed in situ shoot turgor to be estimated in addition to standard water relations traits. Over all years, Female 59 progeny displayed lower osmotic potentials at saturation (Psi(sat)) than Female 63 progeny. Genetic differences in Psi(sat) were similar on both the dry and wet sites. Modulus of elasticity (epsilon) was greater for Female 59 progeny than for Female 63 progeny, producing a compensatory effect resulting in no genetic or site differences in osmotic potential at turgor loss point (Psi(tlp)) or relative water content at turgor loss point (RWC(tlp)). Mean and predawn shoot turgor pressures (P(x) and P(pd)) were higher for Female 59 progeny than for Female 63 progeny and higher at the wet site than the dry site. Genotype x environment trends were observed; compared to Female 63 progeny, Female 59 progeny displayed 9.8 and 5.1% higher P(pd) on the dry and wet sites, respectively, and 3.4 and 9.8% greater P(pd) values in wet and dry years, respectively. Tree volume growth showed no relationship to Psi(tlp) or RWC(tlp), but was correlated with Psi(sat) and P(x); however, the strongest correlation was with P(pd) (r = 0.90). PMID- 12651559 TI - Shoot water relations of mature black spruce families displaying a genotype x environment interaction in growth rate. II. Temporal trends and response to varying soil water conditions. AB - Pressure-volume curves and shoot water potentials were determined for black spruce (Picea mariana (Mill.) BSP) trees from four full-sib families at the Petawawa Research Forest, Ontario, Canada. Trees were sampled from a dry site in 1992 and from the dry site and a wet site in 1993. Modulus of elasticity (epsilon), osmotic potential at turgor loss point (Psi(tlp)) and relative water at turgor loss point (RWC(tlp)) all decreased during the growing season. Osmotic potential at saturation (Psi(sat)) and turgor displayed no general temporal trend. Across a range of environmental conditions, Female 59 progeny had equal or lower Psi(sat), and higher or similar epsilon, mean turgor pressure (P(x)) and predawn turgor pressure (P(pd)) compared with Female 63 progeny. Osmotic potential at saturation decreased as water stress increased from mild to moderate and increased as water stress increased from moderate to severe. Stable genetic differences in Psi(sat) were maintained by the same rate of osmotic adjustment from low to moderate water stress. Modulus of elasticity and RWC(tlp) decreased with decreasing water availability, whereas Psi(tlp) showed no response. The combined effects of Psi(sat) and epsilon resulted in no change in P(pd) as water stress increased from low to moderate values, but turgor declined sharply as water stress increased from moderate to high values. We conclude that drought tolerance traits strongly influence the growth of these black spruce families across sites of varying water availability. PMID- 12651560 TI - Root induction in Pinus ayacahuite by co-culture with Agrobacterium tumefaciens strains. AB - Transformation of in-vitro-derived shoots of Pinus ayacahuite Ehrenb. was achieved by co-culture with an oncogenic strain (A281 x 200) of Agrobacterium tumefaciens. During co-culture rooting also occurred; however, this rooting was not induced by genetic transformation of host cells, because a "disarmed" strain of A. tumefaciens (EHA101) also induced rooting. Furthermore, direct contact between shoots and bacterial cells was not required. Rooting occurred in agar solidified medium and in a soilless substrate (9:1 vermiculite:peat mix). We conclude that A. tumefaciens strains induced rooting in P. ayacahuite through a change in the rhizosphere, probably by producing some root-inducing compound(s), and not through transformation of host cells. PMID- 12651561 TI - Physiological and morphological responses of olive plants to ozone exposure during a growing season. AB - We studied physiological (gas exchange and stomatal aperture) and morphological (individual leaf area and stomatal density) responses in leaves of five-year-old olive plants (Olea europaea L. cvs. Frantoio and Moraiolo) exposed to filtered air containing < 3 ppb O(3) or 100 ppb O(3) for 5 h day(-1) for 120 days in fumigation chambers. After 100 days of treatment, leaf drop and development of necrotic spots were observed in O(3)-fumigated plants of Moraiolo but not of Frantoio. Significant reductions in photosynthetic activity (57%) and stomatal conductance (69%) were detected in O(3)-fumigated plants of Frantoio compared with control plants. In O(3)-fumigated plants of Moraiolo, the decrease in photosynthetic activity (17%) was not statistically significant, but a significant reduction in stomatal conductance (40%) was observed. In both cultivars, leaves that developed after exposure to O(3) showed decreased stomatal aperture (63.6 and 54.8% with respect to the Frantoio and Moraiolo controls, respectively) and one-sided leaf area, and increased stomatal density compared with control leaves. Actual transpiring stomatal surface decreased substantially in both Frantoio (59.8%) and Moraiolo (56.3%) in response to O(3) treatment. Relative transpiring stomatal surface (RTSS) in Frantoio decreased from 0.54 (control) to 0.27% (O(3) treated) of total leaf surface. The corresponding values for Moraiolo were 0.79 and 0.42%. However, because the RTSS of Moraiolo leaves in the O(3) treatment was 0.42 versus 0.27% in Frantoio, the potential uptake of O(3) was higher for Moraiolo plants than for Frantoio plants. The large O(3) induced reduction in transpiring stomatal surface in both cultivars could have significant effects on olive productivity in the Mediterranean area, where high O(3) concentrations persist for long periods during the year. PMID- 12651562 TI - Mechanical injury and fungal infection induce acquired resistance in Norway spruce. AB - Norway spruce trees (Picea abies (L.) Karst.) pretreated by wounding and fungal infection showed highly enhanced resistance to a subsequent challenge inoculation with the pathogenic bluestain fungus Ceratocystis polonica (Siem.) C. Moreau. This is the first time the effectiveness of the constitutive and inducible defenses has been shown to depend on prior wounding and infection in conifers, although such acquired resistance has previously been found in several angiosperms. Trees that were pretreated with a combination of 12 bark wounds (1.6 x 10 cm), four fungal inoculations and four sterile inoculations 1-15 days before mass inoculation with C. polonica at 400 inoculations per square meter over a 0.8 m stem section had significantly shorter necroses in the phloem, less bluestained sapwood, and less dead cambium than untreated control trees. Pretreatment with four fungal or sterile inoculations alone did not lead to enhanced resistance. Pretreatment by bark wounding alone seemed to provide an intermediate degree of resistance compared to bark wounding, fungal inoculations and sterile inoculations combined. All trees had a marked increase in the number of resin ducts in the year of inoculation compared with previous years, suggesting that formation of traumatic resin ducts play an important role in the development and maintainance of enhanced resistance. PMID- 12651563 TI - Elevated CO(2) studies: past, present and future. AB - Increasing concentrations of atmospheric CO(2) are predicted to impact both current and future ecosystems. Elevated CO(2) is also predicted to affect biological processes at many levels of organization. In this overview, we summarize the responses of plants to elevated CO(2) including primary physiological and molecular responses, growth and reproductive responses, effects on plant-plant competition and interactions with other organisms, evolutionary responses, and effects at the ecosystem level. The objectives of this paper are to: (a) overview studies in this issue that were presented at a 1997 meeting entitled "Critical Assessment of the Response of Forest Ecosystems to Elevated Atmospheric Carbon Dioxide," which was sponsored by the Global Change and Terrestrial Ecosystems (GCTE) group of the International Geosphere Biosphere Program (IGBP), (b) review areas of recent progress in CO(2) research, (c) generalize patterns arising from past research, and (d) list critical areas of research for the future. PMID- 12651564 TI - Photosynthetic adjustment in field-grown ponderosa pine trees after six years of exposure to elevated CO(2). AB - Photosynthesis of tree seedlings is generally enhanced during short-term exposure to elevated atmospheric CO(2), but longer-term photosynthetic responses are often more variable because they are affected by morphological, biochemical and physiological feedback mechanisms that regulate carbon assimilation to meet sink demand. To examine biochemical and morphological factors that might regulate the long-term photosynthetic response of field-grown trees to elevated CO(2), we grew ponderosa pine (Pinus ponderosa Dougl. ex Laws.) trees in open-top chambers for six years in native soil at ambient CO(2) (35 Pa) and elevated CO(2) (70 Pa) at a site near Placerville, CA. Trees were well watered and exposed to natural light and ambient temperature. At the end of the sixth growing season at elevated CO(2), net photosynthesis was enhanced 53%, despite reductions in photosynthetic capacity. The positive net photosynthetic response to elevated CO(2) reflected greater relative increases in Rubisco sensitivity compared with the decreases resulting from biochemical adjustments. Analyses of net photosynthetic rate versus internal CO(2) partial pressure curves indicated that reductions in photosynthetic capacity in response to elevated CO(2) were the result of significant reductions in maximum photosynthetic rate (20%), Rubisco carboxylation capacity (36%), and electron transport capacity (21%). Decreased photosynthetic capacity was accompanied by reductions in various photosynthetic components, including total chlorophyll (24%), Rubisco protein content (38%), and mass-based leaf nitrogen concentration (14%). Net photosynthesis was unaffected by morphological adjustments because there was no change in leaf mass per unit area at elevated CO(2). An apparent positive response of photosynthetic adjustment in the elevated CO(2) treatment was the redistribution of N within the photosynthetic system to balance Rubisco carboxylation and electron transport capacities. We conclude that trees, without apparent limitations to root growth, may exhibit photosynthetic adjustment responses in the field after long-term exposure to elevated CO(2). PMID- 12651565 TI - Photosynthetic acclimation to elevated atmospheric CO(2) concentration in the Florida scrub-oak species Quercus geminata and Quercus myrtifolia growing in their native environment. AB - Long-term effects of elevated CO(2) concentration (ambient plus 350 &mgr;mol mol( 1)) on leaf photosynthetic acclimation of two species of a scrub-oak community, Quercus myrtifolia Willd. and Quercus geminata Small, were studied. Plants of both species were grown in open-top chambers in their natural habitat at Kennedy Space Center, Florida, USA. Compared to ambient CO(2), elevated CO(2) stimulated photosynthetic rates by 73 and 51% for Q. geminata and Q. myrtifolia, respectively. Maximum rate of carboxylation (V(cmax)) was significantly reduced by elevated CO(2) in Q. myrtifolia (28%) but not in Q. geminata. Maximum rate of potential electron transport (J(max)) was not significantly reduced by elevated CO(2) in either species. In response to elevated CO(2), specific leaf area decreased in Q. myrtifolia (22%), but not in Q. geminata. Elevated CO(2) caused a significant accumulation of sugars (54%) and starch (264%) in Q. myrtifolia leaves, but not in Q. geminata leaves. Total Rubisco activity in Q. myrtifolia leaves was reduced 40% by elevated CO(2), whereas no significant reduction occurred in Q. geminata leaves. Although both species share a common habitat, they exhibited marked differences in photosynthetic acclimation to elevated CO(2) concentration. PMID- 12651566 TI - Photosynthetic responses of loblolly pine (Pinus taeda) needles to experimental reduction in sink demand. AB - Sink strength in loblolly pine (Pinus taeda L.) was experimentally manipulated on two sun-exposed branches on each of two neighboring trees by excising the emerging terminal cohort (second flush of 1996) during a period of rapid needle expansion. In addition, export of photosynthate was restricted on one of these branches from each tree by removal of bark and phloem just below the second flush of 1995. Treatment-induced changes in needle biochemistry were measured in 3 month-old (first flush of 1996) and 1-year-old (final flush of 1995) needles collected 1, 5 and 8 days after treatment. In 3-month-old needles, sugar concentration increased by 24% one day after leader excision, and increased by 86% on Day 8 after leader excision and girdling. Starch concentration increased by 33% in 3-month-old needles on Day 1 after leader excision, and by 400% in 1 year-old needles on Day 8 after leader excision and girdling. Physiological changes in 3-month-old and 1-year-old needles were measured by open-flow gas exchange and chlorophyll fluorescence on Day 8 after leader excision and girdling. Light- and CO(2)-saturated net photosynthesis decreased following treatment in both 3-month-old and 1-year-old needles (23 and 17%, respectively). Maximum rate of carboxylation (V(cmax)) decreased by 25% in 3-month-old needles and by 31% in 1-year-old needles in response to leader excision and girdling. The combined treatment resulted in a 38% decrease in maximum rate of electron transport (J(max)) in 3-month-old needles and a 37% decrease in J(max) in 1-year old needles. Before leader excision and girdling, 2% oxygen in air stimulated photosynthesis by 17 to 19%, but this stimulation was only 3 to 4% at 9 days after treatment. These physiological responses indicate that experimentally lowered sink strength resulted in rapid feedback inhibition of leaf-level photosynthetic capacity in loblolly pine. PMID- 12651567 TI - Seasonal patterns of photosynthetic light response in Douglas-fir seedlings subjected to elevated atmospheric CO(2) and temperature. AB - Increases in atmospheric CO(2) concentration and temperature are predicted to increase the light response of photosynthesis by increasing light-saturated photosynthetic rates and apparent quantum yields. We examined the interactive effects of elevated atmospheric CO(2) concentration and temperature on the light response of photosynthesis in Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings. Seedlings were grown in sunlit chambers controlled to track either ambient (~400 ppm) CO(2) or ambient + 200 ppm CO(2), at ambient temperature or ambient + 4 degrees C. Photosynthetic light response curves were measured over an 18-month period beginning 32 months after treatments were initiated. Light response curves were measured at the growth CO(2) concentration, and were used to calculate the light-saturated rate of photosynthesis, light compensation point, quantum yield and respiration rate. Elevated CO(2) increased apparent quantum yields during two of five measurement periods, but did not significantly affect light-saturated net photosynthetic rates, light compensation points or respiration rates. Elevated temperature increased all parameters. There were no significant interactions between CO(2) concentration and temperature. We conclude that down-regulation of photosynthesis occurred in the elevated CO(2) treatments such that carbon uptake at a given irradiance was similar across CO(2) treatments. In contrast, increasing temperature may substantially increase carbon uptake rates in Douglas-fir, assuming other environmental factors do not limit photosynthesis; however, it is not clear whether the increased carbon uptake will increase growth rates or be offset by increased carbon efflux through respiration. PMID- 12651568 TI - Direct inhibition of mitochondrial respiratory enzymes by elevated CO(2): does it matter at the tissue or whole-plant level? AB - On average, a doubling in current atmospheric [CO(2)] results in a 15 to 20% direct inhibition of respiration, although the variability associated with this value is large within and among species. Direct effects of CO(2) on respiration may also be relevant to tree canopies because of dynamic fluctuations between nighttime and daytime [CO(2)] throughout the growing season. The mechanism by which CO(2) inhibits respiration is not known. A doubling of ambient [CO(2)] inhibits the activity of the mitochondrial enzymes, cytochrome c oxidase and succinate dehydrogenase. If inhibition of these enzymes is the only factor involved in the direct inhibition of respiration, the overall inhibition of specific respiration will be proportional to the control that such enzymes exert on the overall respiratory rate. We analyzed the effects of [CO(2)] on respiration in an attempt to scale the direct effects of CO(2) on respiratory enzymes to the whole-plant level. Sensitivity analysis showed that inhibition of mitochondrial enzymes by doubling current atmospheric [CO(2)] does not explain entirely the CO(2) inhibition of tissue or whole-plant respiration. We conclude that CO(2)-dependent suppression of respiratory enzymatic activity will be minimal when cytochrome c oxidase inhibition is scaled up from the mitochondria to the whole tree and that the primary mechanism for the direct inhibitory effect remains to be identified. PMID- 12651569 TI - Water relations, stomatal response and transpiration of Quercus pubescens trees during summer in a Mediterranean carbon dioxide spring. AB - Variations in water relations and stomatal response of Quercus pubescens Willd. were analyzed under Mediterranean field conditions during two consecutive summers (1993 and 1994) at two locations characterized by different atmospheric CO(2) concentrations because of the presence at one of them of a CO(2) spring. Trees at the CO(2) spring site have been growing for generations in elevated atmospheric CO(2) concentrations. The heat-pulse velocity technique was used to estimate water use of trees during a 5-month period from June to November 1994. At the end of the sap flow measurements, the trees were harvested and foliage and sapwood area measured. At both sites, maximum leaf conductance was related to predawn shoot water potential. Effects of summer drought on plant water relations, including whole-plant transpiration, were severe, but leaf conductance and water potential recovered to predrought values after major rainfall in September. Leaf conductance, predawn water potential, and sometimes sap flow, decreased in parallel with increases in hydraulic resistance, reaching a minimum in midsummer. Hydraulic resistance was higher in trees at the control site than in trees at the CO(2) spring site. The effect of elevated CO(2) concentration on leaf conductance was less at high leaf-to-air water vapor pressure difference than at low leaf-to air water vapor pressure difference. Mean and diurnal sap fluxes were consistently higher in trees at the control site than in trees at the CO(2) spring site. During the summer period, plant water use varied between the two sites. Trees at the control site had consistently higher sap flow at corresponding values of sapwood cross-sectional area than trees at the CO(2) spring site. Because trees at the CO(2) spring site supported a smaller foliage area for a corresponding sapwood cross-sectional area than trees at the control site, the overall mean sap flux/foliage area ratio did not differ between sites. The results are discussed in terms of effects of elevated CO(2) concentration on plant water use at the organ and whole-tree scale. PMID- 12651570 TI - Seasonal embolism and xylem vulnerability in deciduous and evergreen Mediterranean trees influenced by proximity to a carbon dioxide spring. AB - We investigated how proximity to natural CO(2) springs affected the seasonal patterns of xylem embolism in Quercus ilex L., Quercus pubescens Willd., Fraxinus ornus L., Populus tremula L. and Arbutus unedo L., which differ in leaf phenology and wood anatomy. Xylem embolism was evaluated in both artificially dehydrated branches and in hydrated apical branches collected at monthly intervals during a 20-month sampling period. Initial specific hydraulic conductivity was also evaluated. We found species-dependent differences in xylem hydraulic properties in response to elevated CO(2) concentration. Populus tremula was the most embolized and A. unedo was the least embolized of the species examined. Effects of elevated CO(2) were significant in Q. pubescens, P. tremula and A. unedo, whereas the overall response to elevated CO(2) was less evident in F. ornus and Q. ilex. Specific hydraulic conductivity differed among species but not between sites, although the interaction between species and site was significant. Differences in xylem vulnerability between trees growing near to the CO(2) spring and those growing in control areas were small. Although differences in hydraulic properties in response to elevated CO(2) concentration were small, they may be of great importance in determining future community composition in Mediterranean type forest ecosystems. The possible causes and ecological significance of such differences are discussed in relation to elevated CO(2) concentration and other environmental conditions. PMID- 12651571 TI - Responses of a loblolly pine ecosystem to CO(2) enrichment: a modeling analysis. AB - The development of the Free-Air CO(2) Enrichment (FACE) facilities represents a substantial advance in experimental technology for studying ecosystem responses to elevated CO(2). A challenge arising from the application of this technology is the utilization of short-term FACE results for predicting long-term ecosystem responses. This modeling study was designed to explore interactions of various processes on ecosystem productivity at elevated CO(2) on the decadal scale. We used a forest model (FORDYN) to analyze CO(2) responses-particularly soil nitrogen dynamics, carbon production and storage-of a loblolly pine ecosystem in the Duke University Forest. When a 14-year-old stand was exposed to elevated CO(2), simulated increases in annual net primary productivity (NPP) were 13, 10 and 7.5% in Years 1, 2 and 10, respectively, compared with values at ambient CO(2). Carbon storage increased by 4% in trees and 9.2% in soil in Year 10 in response to elevated CO(2). When the ecosystem was exposed to elevated CO(2) from the beginning of forest regrowth, annual NPP and carbon storage in trees and soil were increased by 32, 18 and 20%, respectively, compared with values at ambient CO(2). In addition, simulation of a 20% increase in mineralization rate led to a slight increase in biomass growth and carbon storage, but the simulated 20% increase in fine root turnover rate considerably increased annual NPP and carbon storage in soil. The modeling results indicated that (1) stimulation of NPP and carbon storage by elevated CO(2) is transient and (2) effects of elevated CO(2) on ecosystem processes-canopy development, soil nitrogen mineralization and root turnover-have great impacts on ecosystem C dynamics. A detailed understanding of these processes will improve our ability to predict long-term ecosystem responses to CO(2) enrichment. PMID- 12651572 TI - Effects of elevated atmospheric CO(2) on phenology, growth and crown structure of Scots pine (Pinus sylvestris) seedlings after two years of exposure in the field. AB - Three-year-old Scots pine (Pinus sylvestris L.) seedlings were grown for two years in the ground in open-top chambers supplied with either an ambient or elevated (ambient + 400 &mgr;mol mol(-1)) CO(2) concentration. Phenological observations and measurements of height and stem diameter growth, absolute and relative growth rates, starch and soluble carbohydrate concentrations of the needles, and crown structure and needle properties were made at frequent intervals throughout the two growing seasons. Elevated CO(2) significantly advanced the date of bud burst in both years. The increase in total needle area in response to elevated CO(2) was accounted for by longer shoots and an increase in individual needle area in the first year, and by an increase in the number and length of shoots in the second year. Stem diameter and tree height were enhanced more by the elevated CO(2) treatment in the first year than in the second, indicating a decreased effect of CO(2) on growth over time. This was confirmed by a study of absolute and relative growth rates of leader shoots. During the first growing season of CO(2) enrichment, mean weekly relative growth rates over the growing season (RGR(m)) were significantly enhanced. During the second year, RGR(m) in ambient CO(2) closely matched that in elevated CO(2). PMID- 12651573 TI - Influence of increased atmospheric CO(2) concentration on quality of plant material and litter decomposition. AB - Nitrogen (N) and lignin concentrations in plant tissues and litter of plants grown in greenhouses or open-top chambers in elevated atmospheric CO(2) concentration were compared with those of plants grown in ambient air in short term studies. We also compared the N concentration of plant material of Quercus ilex L. and Q. pubescens Willd. growing in the vicinity of natural CO(2)-springs with that of the same species growing at a control site. In the short-term studies, elevated CO(2) caused significant decreases in tissue N concentration and the extent of the decrease varied with species. Nitrogen amendment of the soil lessened the CO(2)-enrichment effect. Lignin concentration was modified by elevated CO(2) and the effect was species specific, but no general positive or negative trend was evident. A comparison of trees growing under natural conditions near a natural CO(2)-spring and at a control site revealed no site differences in N concentration of the plant material. A comparison of published results on decomposition rates of litter produced in elevated atmospheric CO(2) and in ambient air indicated that CO(2) enrichment can cause both enhancements and decreases of carbon mineralization. We conclude that (1) long-term responses to elevated CO(2) could differ from the results obtained from short-term studies and that (2) biodiversity could be an important factor altering the sign of the feedback on atmospheric CO(2) concentration. We also discuss the implications of our finding of a long-term, inhibitory effect of the initial N concentration of litter on the decomposition rate of litter and its consequence on ecosystem feedback. PMID- 12651574 TI - Rhizosphere feedbacks in elevated CO(2). AB - Understanding rhizosphere processes in relation to increasing atmospheric CO(2) concentrations is important for predicting the response of forest ecosystems to environmental changes, because rhizosphere processes are intimately linked with nutrient cycling and soil organic matter decomposition, both of which feedback to tree growth and soil carbon storage. Plants grown in elevated CO(2) substantially increase C input to the rhizosphere. Although it is known that elevated CO(2) enhances rhizosphere respiration more than it enhances root biomass, the fate and function of this extra carbon input to the rhizosphere in response to elevated CO(2) are not clear. Depending on specific plant and soil conditions, the increased carbon input to the rhizosphere can result in an increase, a decrease, or no effect on soil organic matter decomposition and nutrient mineralization. Three mechanisms may account for these inconsistent results: (1) the "preferential substrate utilization" hypothesis; (2) the "priming effect" hypothesis; and (3) the "competition" hypothesis, i.e., competition for mineral nutrients between plants and soil microorganisms. A microbial growth model is developed that quantitatively links the increased rhizosphere input in response to elevated CO(2) with soil organic matter decomposition. The model incorporates the three proposed mechanisms, and simulates the complexity of the rhizosphere processes. The model also illustrates mechanistically the interactions among nitrogen availability, substrate quality, and microbial dynamics when the system is exposed to elevated CO(2). PMID- 12651575 TI - Simulated nitrogen cycling response to elevated CO(2) in Pinus taeda and mixed deciduous forests. AB - Interactions between elevated CO(2) and N cycling were explored with a nutrient cycling model (NuCM, Johnson et al. 1993, 1995) for a Pinus taeda L. site at Duke University, North Carolina, and a mixed deciduous site at Walker Branch, Tennessee. The simulations tested whether N limitation would prevent growth increases in response to elevated CO(2), and whether growth responses to CO(2) in N-limited systems could be facilitated by increasing the biomass/N ratio (reducing N concentration) or increasing litter N mineralization, or both. Nitrogen limitation precluded additional growth when target growth rates and litterfall were increased (simulating potential response to elevated CO(2)) at the Duke University site. At the Walker Branch site, increasing target growth and litterfall caused a 7% increase in growth. Reducing foliar N concentrations reduced growth because of N limitation created by reduced litter quality (C:N ratio), reduced decomposition and increased N accumulation on the forest floor. These effects were most pronounced at the Duke University site, because the forest floor N turnover rate was lower than at the Walker Branch site. Reducing wood N concentration allowed prolonged increases in growth because of greater biomass/N; however, N uptake was reduced, allowing greater N immobilization on the forest floor and in soil. Increased N mineralization caused increased growth at the Duke University site, but not at the Walker Branch site. These simulations pose the counterintuitive hypothesis that increased biogeochemical cycling of N (as a result of increased litterfall N) causes reduced growth in an N-limited system because of increased accumulations of N on the forest floor and in soil. Translocation of N from senescing leaves before litterfall mitigates this response by allowing the trees to retain a greater proportion of N taken up rather than recycle it back to the forest floor and soil where it can be immobilized. Eliminating N translocation at Walker Branch changed the direction as well as the magnitude of the responses in three of the four scenarios simulated. Because the NuCM model does not currently allow translocation in coniferous species, the effects of translocation on N cycling in the Duke University simulations are not known. PMID- 12651576 TI - Tissue chemistry and carbon allocation in seedlings of Pinus palustris subjected to elevated atmospheric CO(2) and water stress. AB - Longleaf pine (Pinus palustris Mill.) seedlings were grown in 45-l pots and exposed to ambient or elevated (365 or 730 &mgr;mol CO(2) mol(-1)) CO(2) concentration in open-top chambers for 20 months. Two water-stress treatments (target values of -0.5 or -1.5 MPa xylem pressure potential) were imposed 19 weeks after initiation of the study. At harvest, tissues (needles, stems, taproots, coarse roots, and fine roots) were analyzed for carbon (C), nitrogen (N), nonpolar extractives (fats, waxes, and oils), nonstructural carbohydrates (sugars and starch), structural components (cellulose and lignin), and tannins. The greatest dry weights and lowest N concentrations occurred in tissues of plants grown at elevated CO(2) or with adequate water. Although allocation of C fractions among tissues was generally unaffected by treatments, concentrations of the analyzed compounds were influenced by treatments in needles and taproots, but not in stems and lateral roots. Needles and taproots of plants exposed to elevated CO(2) had increased concentrations of nonstructural carbohydrates. Among plant tissues, elevated CO(2) caused reductions in structural C concentrations and foliar concentrations of fats, waxes and oils. PMID- 12651577 TI - Estimating stem and root-anchorage flexibility in trees. AB - This paper describes a nondestructive method for distinguishing root flexibility from stem flexibility in living trees. It is used here for Sitka spruce (Picea sitchensis (Bong.) Carriere.), but is applicable to any species where the main stem is normally straight and near-vertical. Well-known engineering equations permit the calculation of deflected shape for a vertical cantilever with arbitrary distribution of mass and bending stiffness, when infjected to a lateral force. The equations are used to calculate stem deflections of four Sitka spruce trees for which the stem and branch mass distribution and stem taper have been measured. Free parameters in the mathematical model are a nominal value of Young's Modulus E (assumed uniform and isotropic over the cross section and height of the tree stem) and a root-anchorage stiffness k. The former allows the stem to curve, whereas the latter represents the flexibility of the roots and allows the stem to tilt elastically at ground level. For each of the four trees, the calculated deflection curve is compared with actual deflections measured when the living tree is pulled by a rope at a specified point. By adjusting both E and k, iteratively, a best fit solution is obtained. This provides a simple and effective way to determine both stem stiffness and root hinge stiffness from a single experiment on a living tree. PMID- 12651578 TI - Ecophysiology of Cecropia schreberiana saplings in two wind regimes in an elfin cloud forest: growth, gas exchange, architecture and stem biomechanics. AB - The hypothesis that the short, stunted development of elfin cloud forest trees is influenced by strong winds was tested by studying growth, gas exchange and biomechanics of potted saplings of Cecropia schreberiana Miq. ex. C. peltata L. (Cecropiaceae) exposed to two natural wind regimes (exposed and protected) at high elevation sites in the Luquillo Experimental Forest, Puerto Rico. The wind exposure treatment produced several thigmomorphogenetic responses, including reductions in plant stature and crown area, changes in allocation patterns, and increased root to shoot ratio, leaf abrasion and leaf epinasty. Wind-exposure decreased maximum photosynthetic rate and respiration on an area basis, but not on a leaf-mass basis. Wind-exposed plants had lower apparent quantum yields, and higher light compensation points than wind-protected plants. Photosynthetic nitrogen-use efficiency was lower in wind-exposed plants, but such plants had higher leaf nitrogen concentration than wind-protected plants. There were no effects of treatments on stomatal conductance, transpiration rate and water-use efficiency. Stems of wind-exposed plants had lower second moment of area, apparent modulus of elasticity, flexural stiffness and stem density, but higher water content than wind-protected plants. Tissue-density-specific stiffness and the calculated critical height were not affected by the treatments. Wind-exposed plants were biomechanically less predisposed to bending and failing under their own weight than wind-protected plants because their safety factors were smaller, indicating that maintenance of an ontogenetically less developed structure enables plants to cope with wind loading. Windward trees showed a lower scaling component of the allometric relationship between diameter and height than leeward trees. PMID- 12651579 TI - Evidence that hydraulic conductance limits photosynthesis in old Pinus ponderosa trees. AB - We tested the hypotheses that hydraulic conductance is lower in old (about 250 years old and 30 m tall) compared to young (about 40 years old and 10 m tall) Pinus ponderosa Dougl. ex Laws. trees and that lower hydraulic conductance of old trees limits their photosynthesis. Hydraulic conductance at the end of summer 1995, calculated from leaf water potential and leaf gas exchange measurements on one-year-old needles, was 44% lower in old trees compared to young trees growing in a mixed age-class stand on the east slope of the Oregon Cascades. Whole-tree sapflow per unit leaf area averaged 53% lower in old trees compared to young trees and mean hydraulic conductance calculated from sapflow and water potential data was 63% lower in old trees than in young trees. For the entire summer, stomatal conductance (g(s)) and assimilation (A) declined more steeply with air saturation deficit (D) in old trees than in young trees. For both old and young trees, mean g(s) and A were approximately 32 and 21% lower, respectively, at typical midday D values (2.5-3.0 kPa). We hypothesized that if hydraulic conductance limits g(s) and A, then increasing or decreasing the leaf specific conductance of a branch will result in proportional changes in the responses of g(s) and A with D. Removal of 50% of the foliage from a set of experimental branches on old trees caused g(s) and A to decline less steeply with D in early summer, but values were not significantly different from control values in late summer. Cutting transverse notches in branches on young trees had no effect on the responses of g(s) and A with D. Leaf nitrogen content and photosynthetic capacity were similar suggesting that differences in g(s) and A between old and young trees were not caused by differences in photosynthetic capacity. PMID- 12651580 TI - Influence of canopy light environment and nitrogen availability on leaf photosynthetic characteristics and photosynthetic nitrogen-use efficiency of field-grown nectarine trees. AB - Relationships between CO(2) assimilation at light saturation (A(max)), nitrogen (N) content and weight per unit area (W(A)) were studied in leaves grown with contrasting irradiances (outer canopy versus inner canopy) and N supply rates in field-grown nectarine trees Prunus persica L. Batsch. cv. Fantasia. Both A(max) and N content per unit leaf area (N(A)) were linearly correlated to W(A), but leaves in the high-N treatment had higher N(A) and A(max) for the same value of W(A) than leaves in the low-N treatment. The curvilinear relationship between photosynthesis and total leaf N was independent of treatments, both when expressed per unit leaf area A(maxA) and N(A)) and per unit leaf weight (A(maxW) and N(W)), but the relationship was stronger when data were expressed on a leaf area basis. Both A(maxA) and N(A) were higher for outer canopy leaves than for inner canopy leaves and A(maxW) and N(W) were higher for leaves in the high-N treatment than for leaves in the low-N treatment. The relationship between A(max) and N resulted in a similar photosynthetic nitrogen-use efficiency at light saturation (A(max)NUE) for both N and light treatments. Photosynthetic nitrogen use efficiency was similar among treatments throughout the whole light response curve of photosynthesis. Leaves developed in shade conditions did not show higher N-use efficiency at low irradiance. At any intercellular CO(2) partial pressure (C(i)), photosynthetic CO(2) response curves were higher for outer canopy leaves and, within each light treatment, were higher for the high-N treatments than for the low-N treatments. Consequently, most of the differences among treatments disappeared when photosynthesis was expressed per unit N. However, slightly higher assimilation rates per unit N were found for outer canopy leaves compared with inner canopy leaves, in both N treatments. Because higher daily irradiance within the canopies of the low-N trees more than compensated for the lower photosynthetic performances of these leaves compared to the leaves of high-N trees, daily carbon gain (and N-use efficiency on a daily assimilation basis) per leaf was higher for the low-N treatment than for the high-N treatment in both outer and inner canopy leaves. PMID- 12651581 TI - Spatial distribution of leaf dry weight per area and leaf nitrogen concentration in relation to local radiation regime within an isolated tree crown. AB - To assess the spatial distribution of photosynthetic capacity within an isolated 20-year-old walnut tree (Juglans regia L.) crown, the distribution of relevant leaf characteristics was measured. Variations in leaf dry weight per area (W(a)), and nitrogen content on a weight (N(w)) and area basis (N(a)) were studied along two horizontal and one vertical gradients of leaf irradiance, at two dates (July 30 and September 3). In addition, the content of total nonstructural carbon on a weight (TNC(w)) and area basis (TNC(a)) was measured on July 30. Concurrently, the spatial distribution of daily integrated leaf irradiance within the crown was simulated by a three-dimensional radiation transfer model over a one week period before sampling at each date. High spatial heterogeneity was observed for W(a) (from 50 to 140 g m(-2)), TNC(a) (from 4 to 17 g m(-2)) and N(a) (from 1.2 to 3.6 g m(-2)) among the foliage. Although TNC(w) and N(w) were not correlated and only weakly correlated to daily leaf irradiance, respectively, W(a), TNC(a) and N(a) were strongly correlated to daily leaf irradiance. The relationship between observed N(a) and simulated daily leaf irradiance was used to assess the spatial distribution of N(a) within the crown at each date. Total leaf nitrogen in the foliage was estimated to be 339 g in late July and 317g in early September. For the whole crown (i.e., 1729 current-year shoots), N(a) increased strongly with basal shoot diameter (an index of "shoot vigor"), highlighting the fact that large shoots were mainly located in sunlit locations and exhibited high photosynthetic capacity. PMID- 12651582 TI - Nutrient uptake by intact mycorrhizal Pinus sylvestris seedlings: a diagnostic tool to detect copper toxicity. AB - We developed a nondestructive method for detecting early toxic effects of inflethal copper (Cu) concentrations on ectomycorrhizal and non-mycorrhizal (NM) Scots pine (Pinus sylvestris L.) seedlings. The fungal symbionts examined were Paxillus involutus (Fr.) Fr., Suillus luteus (Fr.) S.F. Gray and Thelephora terrestris (Ehrh.) Fr. The accumulation of Cu in needles and fungal development (ergosterol) in roots and infstrate were assessed. Inorganic phosphate (P(i)) and ammonium (NH(4) (+)) uptake capacities were determined in a semi-hydroponic cultivation system on intact P-limited plants that were exposed for 3 weeks to 0.32 (control), 8 or 16 &mgr;moles Cu(2+). Short-term effects of a 1-hour exposure to 32 &mgr;moles Cu(2+) on nutrient uptake rates were also determined. None of the Cu(2+) treatments affected plant growth or root ergosterol concentrations. The active fungal biomass in infstrate invaded by S. luteus was reduced by 50% in the 16 &mgr;M Cu(2+) treatment compared with the control treatment; however, colonization by S. luteus prevented an increased accumulation of Cu in the needles. In contrast, the 16 &mgr;M Cu(2+) treatment caused a 2.2 fold increase in needle Cu concentration in NM plants. Ergosterol concentrations in the infstrate colonized by P. involutus and T. terrestris were not affected by 16 &mgr;molar Cu(2+). Although P. involutus and T. terrestris were less sensitive to Cu(2+) than S. luteus, T. terrestris did not prevent the accumulation of Cu in needles of its host plant in the 16 &mgr;molar Cu(2+) treatment. Mycorrhizal plants consistently had higher P(i) and NH(4) (+) uptake capacities than NM plants. In the control treatment, specific P(i) uptake rates were almost 10, 4 and 3 times higher in plants associated with P. involutus, S. luteus and T. terrestris, respectively, than in NM plants, and specific NH(4) (+) uptake rates were about 2, 2 and 5 times higher, respectively, than those of NM seedlings. Compared with the corresponding control plants, a 3-week exposure to 8 &mgr;M Cu(2+) had no effect on the nutrient uptake potential of plants. In contrast, the 16 &mgr;M Cu(2+) treatment significantly reduced P(i) uptake capacity of all plants and decreased NH(4) (+) uptake capacity of seedlings colonized by S. luteus or T. terrestris. The 32 &mgr;M Cu(2+) 1-h shock treatment reduced specific NH(4) (+) and P(i) uptake rates of roots colonized by S. luteus to 39 and 77%, respectively, of the original rates. The Cu(2+) 1-h shock treatment reduced the NH(4) (+) uptake rate of NM plants by 51%. PMID- 12651583 TI - Nutrition and bud removal affect biomass and nutrient allocation in Douglas-fir and western red cedar. AB - Seedlings of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) and western red cedar (Thuja plicata J. Donn ex D. Don) were grown at high (250 mg l(-1)) and low (20 mg l(-1)) nitrogen (N) supply for a year. Before the second growing season, half of the seedlings in each nutrient treatment were allocated to the other treatment. Half of the seedlings in each nutrient treatment then had all growing points removed. Biomass and N, phosphorus (P) and potassium (K) concentrations of old and new shoots and roots were measured three times in the second year to test the interaction of current-year and previous-year nutrient supply on biomass and nutrient allocation in these two species with different growth habits. Pruned seedlings served as controls. Unpruned seedlings of both species increased in height throughout the second growing season, except for Douglas-fir in the N250 - > N20 treatment. Repeated pruning did not prevent new shoot growth, but resulted in a 12 to 52% reduction in biomass of new shoots and new and old roots. Seedlings receiving a low N supply in the first growing season were more severely affected by pruning than seedings receiving a high N supply. Growth was reduced more by pruning in western red cedar than in Douglas-fir. Concentrations of N, P and K were higher in pruned seedlings than in unpruned seedlings. Although dry weights of all plant parts in all treatments increased throughout the second growing season, some retranslocation of N, P and K was observed from old shoots of both species in the N250 --> N20 and N20 --> N20 treatments after August. Quantities of N, P and K retranslocated were greatest in seedlings grown the previous year in the high-N treatment. PMID- 12651584 TI - Effects of nitrogen source on growth and activity of nitrogen-assimilating enzymes in Douglas-fir seedlings. AB - Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings were grown for 68 days in a growth chamber in nutrient solutions with ammonium, nitrate or ammonium nitrate as the nitrogen source. Among the nitrogen sources tested, whole-seedling biomass, relative growth rate (RGR), root and shoot elongation, and number of lateral roots, were greatest in seedlings grown with ammonium. In the absence of nitrogen, plant growth and formation of lateral roots were poor. Initially, glutamine synthetase, NAD-glutamate dehydrogenase and aspartate aminotransferase activities were high in young roots and shoots, but all three enzymatic activities decreased after one month of culture. In root apices, glutamine synthetase and aspartate aminotransferase activities were higher than NAD glutamate dehydrogenase activity. Enzymatic activities were often higher in ammonium-fed seedlings than in seedings supplied with the other forms of nitrogen. Activities of all three enzymes were significantly reduced in seedlings grown in the absence of nitrogen. The beneficial effect of ammonium is discussed on the basis of its involvement in the assimilation pathways of Douglas-fir. PMID- 12651585 TI - Predisposition of trees to drought stress by ozone. AB - This paper reviews research on the effects of ozone on stomatal behavior, and draws attention to the direct link between stomatal function and air pollution. Contradictory results concerning the effects of air pollutants on stomatal activity are attributed to the use of univariate statistical treatments of data. Alternative experimental methods that subject stomata to a functional test are described. These methods verify the hypothesis that ozone predisposes trees to drought stress. It is proposed that ozone directly attacks the walls of guard and subsidiary cells, leading to impairment of the sensory mechanism of the stomata. PMID- 12651586 TI - Seasonal changes in photosynthesis of trees in the flooded forest of the Mapire River. AB - We studied the flood tolerance of five tree species growing in the flooded forest adjacent to the Mapire river, in SW Venezuela. Mean photosynthetic rate and leaf conductance were 11 &mgr;mol m(-2) s(-1) and 700 mmol m(-2) s(-1), respectively. Xylem water potential ranged from -0.08 to -1.15 MPa. Based on leaf gas exchange as a criterion of tolerance to flooding, two response patterns were identified: (1) decreasing photosynthetic rate with increasing flooding and leaf conductance (Psidium ovatifolium Berg. ex Desc., Campsiandra laurifolia Benth., Symmeria paniculata Benth. and Acosmium nitens (Vog.) Benth); and (2) independence of photosynthesis and leaf conductance from flooding (Eschweilera tenuifolia (Berg.) Miers.). In the first response pattern, declining photosynthetic rate with flooding may be interpreted as a sign of reduced flood tolerance, whereas the second response pattern may indicate increased flood tolerance. An increase in xylem water potential with depth of water column was found for all species (with the possible exception of P. ovatifolium), indicating that flooding does not cause water stress in these trees. Submerged leaves that had been under water for between four days and four months generally had photosynthetic rates and leaf conductances similar to those of aerial leaves, indicating maintenance of photosynthetic capacity under water. Daily positive oscillations in glucan content in submerged leaves of P. ovatifolium and C. laurifolia suggest that submerged leaves do not represent a sink for photosynthates produced by aerial leaves. PMID- 12651587 TI - Thinning, fertilization, and crown position interact to control physiological responses of loblolly pine. AB - To examine physiological responses to thinning, fertilization, and crown position, we measured net photosynthesis (P(n)), transpiration (E), vapor pressure difference (VPD), stomatal conductance (g(s)), and xylem pressure potential (Psi(1)) between 0930 and 1130 h under ambient conditions in the upper and lower crowns of a 13-year-old loblolly pine (Pinus taeda L.) plantation six years (1994) after the treatments were applied. Photosynthetic photon flux density (PPFD) and air temperature (T(a)) within the canopy were also recorded. Needle P(n) of thinned trees was significantly enhanced by 22-54% in the lower crown, because canopy PPFD increased by 28-52%. Lower crown foliage of thinned plots also had higher E and g(s) than foliage of unthinned plots, but thinning had no effect on needle Psi(1) and predawn xylem pressure potential (0430-0530 h; Psi(pd)). Tree water status did not limit P(n), E and g(s) during the late morning measurements. Fertilization significantly decreased within-canopy PPFD and T(a). Needle Psi(1) was increased in fertilized stands, whereas P(n), E and g(s) were not significantly altered. Upper crown foliage had significantly greater PPFD, P(n), VPD, g(s), E, and more negative Psi(1) than lower crown foliage. In both crown positions, needle P(n) was closely related to g(s), PPFD and T(a) (R(2) = 0.77 for the upper crown and 0.82 for the lower crown). We conclude that (1) silvicultural manipulation causes microclimate changes within the crowns of large trees, and (2) needle physiology adjusts to the within-crown environmental conditions. PMID- 12651588 TI - Seasonal dynamics of electrical impedance parameters in shoots and leaves related to rooting ability of olive (Olea europea) cuttings. AB - Electrical impedance parameters were measured in shoots and leaves of Olea europaea L. for 18 months to determine seasonal variations in intracellular and extracellular resistances and in the state of membranes; these factors were related to rooting ability. Double- and single-DCE (ZARC) models were used as equivalent circuits for shoots and leaves, respectively. Seasonal variations were observed in all of the impedance parameters measured. Intracellular resistance of the shoots increased during the winter resting period, whereas intracellular resistance of the leaves decreased. Relaxation times for both leaves and shoots decreased during the winter. Close relationships were found between rooting ability and intracellular and extracellular resistances and relaxation times of shoots and leaves. PMID- 12651589 TI - Carbohydrate metabolism of vegetative and reproductive sinks in the late-maturing peach cultivar 'Encore' AB - Activities of NAD(+)-dependent sorbitol dehydrogenase (SDH), sorbitol oxidase (SOX), sucrose synthase (SS), acid invertase (AI), and neutral invertase (NI) in 'Encore' peach (Prunus persica L.) fruits and developing shoot tips were assayed during the growing season to determine whether carbohydrate metabolizing enzymes could serve as indicators of sink strength. In fruit flesh, SS activity was detected during Stage I of growth, when cells were actively dividing, and SDH activity was detected during Stage III, when cells were actively enlarging. Acid invertase activity was detected during Stage I and showed a closer correlation with relative increase in fruit weight during the growing season than SS activity. During seed filling and pit hardening (Stage II), when relative fruit growth rate was slowest, activities of carbohydrate metabolizing enzymes in fruit flesh were not detectable. No SOX activity was detected during Stages I and II. The highest sucrose content occurred near the end of fruit development when the activities of sucrose metabolizing enzymes were low. In developing shoot tips, the sorbitol:sucrose ratio was 2:1 (w/w) and SDH activity was low at the beginning and end of the season when vegetative growth was slowest. The sorbitol:sucrose ratio changed to 1:1 (w/w) along with an increase in SDH activity in shoot tips during the mid-growing season. In 'Nemaguard' peach, SDH exhibited higher activity in root tips than in other organs. Among the sorbitol- and sucrose-metabolizing enzyme activities, only SDH activity was positively correlated with shoot growth in 'Nemaguard' plants. PMID- 12651590 TI - Growth and nutrition of Betula pendula at different relative supply rates of zinc. AB - Small birch plants (Betula pendula Roth) were cultivated in a hydroponic spray solution where the relative growth rate (R(G); day(-1)) was controlled by the relative supply rate of zinc (R(Zn); day(-1)). After an adjustment phase to steady-state growth, R(G) equaled R(Zn). The R(Zn) treatments were 0.05, 0.125 and 0.20 day(-1) with free access to all other nutrients. In an additional treatment, there was free access to all nutrients, including zinc (FA treatment). The pH of the nutrient solution was approximately 4.5 and conductivity was 100 &mgr;S cm(-1). The duration of each treatment depended on R(Zn) and ranged from 4 (FA treatment) to 10 weeks (at R(Zn) = 0.05 day(-1)). The plants showed persistent and typical zinc-deficiency symptoms at steady-state growth when R(G) was 0.05 and 0.125 day(-1), whereas there were few symptoms when R(G) was 0.2 day(-1). The Zn concentration of the plants ranged from 8 (at R(Zn) = 0.05 day( 1)) to 21 &mgr;g g(DM) (-1) (DM = dry mass) (at R(Zn) = 0.2 day(-1)) and was approximately 42 &mgr;g g(DM) (-1) in the FA treatment. Uptake rates of Zn, calculated per root growth rate (&mgr;mol g(DM, root) (-1)), were about 2.8 times higher at R(Zn) = 0.20 day(-1) than at R(Zn)= 0.05 day(-1). The root and stem biomass fractions were approximately constant at all supply rates of Zn, whereas the leaf biomass fraction tended to increase with increasing supply rate of Zn. Net assimilation rate was constant from FA to an R(Zn) of 0.125 day(-1) but decreased by a factor of about 2 at R(Zn) = 0.05 day(-1). Leaf area ratio and specific leaf area were smaller at low supply rates of zinc than at high supply rates. PMID- 12651591 TI - Leaf gas exchange and cholorphyll fluorescence in relation to leaf angle, azimuth, and canopy position in the tropical pioneer tree, Macaranga conifera. AB - We tested the hypothesis that, in tropical pioneer tree species, vertical leaf angle contributes to high carbon gain because it minimizes damage caused by high irradiances. Diurnal changes in leaf gas exchange and chlorophyll fluorescence were measured in east-facing (EL), west-facing (WL) leaves, and in leaves artificially held horizontal (HL) in the uppermost canopy of Macaranga conifera (Zoll.) Muell. Arg. Maximum values of net photosynthetic rate (P(n)) for EL and HL reached 12 &mgr;mol m(-2) s(-1), whereas maximum P(n) for WL was only 6 &mgr;mol m(-2) s(-1). Midday depressions of P(n) and stomatal conductance occurred at high photosynthetic photon flux densities (PPFD), especially for HL. Photosystem II quantum yield (DeltaF/F(m)') of HL for a given PPFD at the leaf surface was lower in the afternoon than in the morning. Values of DeltaF/F(m)' for HL measured at dusk were lower than those measured just before dawn, suggesting that HL suffered from high light and heat load. Variations in the morphology and physiology of the canopy leaves were associated with different light environments, and there was circumstantial evidence of a transitional point at a PPFD of about 20-30% of full sunlight. Maximum P(n) and nitrogen (N) content were higher in upper canopy leaves than in lower canopy leaves, and the differences were mainly associated with differences in lamina thickness. We conclude that the vertical leaf angle and thick lamina of the top canopy leaves contributed to enhance total carbon gain of the whole plant. PMID- 12651592 TI - Mapping tree root systems with ground-penetrating radar. AB - A ground-penetrating radar (GPR) technique was used to study the three dimensional distribution of root systems of large (DBH = 14 to 35 cm) oak trees (Quercus petraea (Mattusch.) Liebl.) in relatively dry, luvisoil on loamy deluvium and weathered granodiorite. We used a pulse EKKO 1000 GPR system, a profile grid of 0.25 x 0.25 meters, at 0.05 m intervals, and a signal frequency of 450 MHz, to assure resolution of about 3 cm in both directions (further increases in resolution up to 1 cm are possible with the system). Coarse root density was 6.5 m m(-2) of stand area and 3.3 m m(-3) of soil volume. Maximum rooting depth of the experimental oaks was 2 m, and the root ground plan was significantly larger (about 1.5 times) than the crown ground plan. Based on earlier studies of Quercus robur L. from floodplain forests, where the extent of the root systems was much smaller (root ground plan:crown ground plan ratio of 0.6), we conclude that the high root ground plan:crown ground plan ratio indicates less favorable conditions of water supply at the experimental site than in the floodplain forest. The ground-penetrating radar system is noninvasive and allows relatively rapid and repeated measurements of the distribution of coarse root systems of trees. PMID- 12651593 TI - Variation in lateral shoot elongation patterns and hybrid vigor in full-sib families and interspecific hybrids of larch. AB - Time of shoot initiation and cessation, and rate and duration of lateral shoot elongation of full-sib families of Larix decidua Mill., L. laricina (Du Roi) C. Koch, and interspecific hybrids between L. decidua and L. kaempferi (Lamb.) Carriere, L. kaempferi x decidua, L. laricina x decidua, and L. laricina x kaempferi were explored in relation to hybrid vigor determined by height growth after 5 years. Height growth of L. laricina and L. decidua families was significantly less than height growth of the interspecific hybrids. Differences were observed for all lateral shoot elongation traits among L. laricina, L. decidua, and the interspecific hybrids. Lateral shoots of L. laricina families were the first to both initiate and cease elongation, and grew for the shortest period of time. Overall, the length of the shoot elongation period and date of cessation of shoot growth were significantly correlated with total height growth after 5 years, but the time of shoot initiation was not. Rate of elongation of lateral shoots was strongly correlated with growth increment in 1996, but was weakly correlated with total height growth after 5 years. Hybrid vigor in Larix seems to be partly a function of late cessation or increased duration of shoot elongation, or both. PMID- 12651596 TI - Aberrant DNA methylation: have we entered the era of more than one type of colorectal cancer? PMID- 12651595 TI - Differential regulation of EphA2 in normal and malignant cells. PMID- 12651597 TI - Kruppel cripples prostate cancer: KLF6 progress and prospects. PMID- 12651598 TI - Genetic and biological subgroups of low-stage follicular thyroid cancer. AB - Investigations of cancer-specific gene rearrangements have increased our understanding of human neoplasia and led to the use of the rearrangements in pathological diagnosis of blood cell and connective tissue malignancies. Here, we have investigated 3p25 rearrangements of the peroxisome proliferator-activated receptor gamma (PPAR gamma) gene in follicular epithelial tumors of the human thyroid gland. Eleven of 42 (26%) low-stage follicular carcinomas, 0 of 40 follicular adenomas, 1 of 30 Hurthle cell carcinomas, 1 of 90 papillary carcinomas, and 0 of 10 nodular goiters had 3p25 rearrangements by interphase fluorescence in situ hybridization. All 11 follicular carcinomas with 3p25 rearrangement exhibited strong, diffuse nuclear immunoreactivity for PPAR gamma, consistent with expression of PPAR gamma fusion protein. Twelve of 42 (29%) low stage follicular carcinomas had 3p25 aneusomy without PPAR gamma rearrangement (P = 0.01), suggesting that PPAR gamma rearrangement and aneuploidy are independent early events in follicular cancer. Eleven of 12 follicular carcinomas with 3p25 aneusomy exhibited no PPAR gamma immunoreactivity, supporting the existence of two independent pathways. Follicular carcinoma patients with PPAR gamma rearrangement more frequently had vascular invasion (P = 0.01), areas of solid/nested tumor histology (P < 0.001), and previous non-thyroid cancers (P < 0.01) compared with follicular carcinoma patients without PPAR gamma rearrangement. Our experiments identify genetic subgroups of low-stage follicular thyroid cancer and provide evidence that follicular carcinomas with PPAR gamma rearrangement are a distinct biological entity. The findings support a model in which separate genetic alterations initiate distinct pathways of oncogenesis in thyroid carcinoma subtypes. PMID- 12651599 TI - Mononuclear cell-infiltrate inhibition by blocking macrophage-derived chemokine results in attenuation of developing crescentic glomerulonephritis. AB - Glomerular monocyte/macrophage (Mo/M phi) infiltrates play a role in many forms of glomerulonephritis (GN), and the intensity of Mo/M phi trafficking correlates with the loss of renal function and histological damage. We analyzed the functional role of macrophage-derived chemokine (MDC), a potent mononuclear cell chemoattractant, during the progression of anti-glomerular basement membrane (GBM) antibody (Ab) GN, a model of crescentic GN in the WKY rat, and whether the effects of MDC were dependent on its receptor CCR4. MDC mRNA and protein expression were markedly induced in nephritic glomeruli throughout the disease. Blocking the function of MDC did not affect the developing of the disease from days 2 to 7, but it dramatically blocked M omicron/M phi infiltration in the glomeruli, prevented crescent formation, and reversed renal function impairment during days 7 to 14 of the anti-GBM GN. In this study, we also found that MDC activity on M omicron/M phi in this GN was at least partly dependent on a new variant of CCR4. These results suggest that MDC is critically involved in the development of anti-GBM GN from acute glomerular injury to irreversible tissue damage. In addition, an antagonist to MDC may represent a prime drug target for therapeutic application to intervene in the progression of anti-GBM GN and in other M omicron/M phi-dominant GN. PMID- 12651600 TI - CD21-positive follicular dendritic cells: A possible source of PrPSc in lymph node macrophages of scrapie-infected sheep. AB - Natural sheep scrapie is a prion disease characterized by the accumulation of PrP(Sc) in brain and lymphoid tissues. Previous studies suggested that lymph node macrophages and follicular dendritic cells (FDC) accumulate PrP(Sc). In this study, lymph nodes were analyzed for the presence of PrP(Sc) and macrophage or FDC markers using dual immunohistochemistry. A monoclonal antibody (mAb) to the C terminus of PrP reacted with CD172a+ macrophages and CD21+ FDC processes in secondary follicles. However, a PrP N-terminus-specific mAb reacted with CD21+ FDC processes but not CD172a+ macrophages in secondary follicles. Neither the PrP N-terminus nor C-terminus-specific mAb reacted with CD172a+ macrophages in the medulla. These results indicate that lymph node follicular macrophages acquire PrP(Sc) by phagocytosis of CD21+ FDC processes. The results also suggest that follicular macrophages have proteases that process full-length PrP(Sc) to N terminally truncated PrP(Sc). PMID- 12651601 TI - Platelet-derived growth factor-B enhances glioma angiogenesis by stimulating vascular endothelial growth factor expression in tumor endothelia and by promoting pericyte recruitment. AB - Platelet-derived growth factor (PDGF)-B and its receptor (PDGF-R) beta are overexpressed in human gliomas and responsible for recruiting peri-endothelial cells to vessels. To establish the role of PDGF-B in glioma angiogenesis, we overexpressed PDGF-B in U87MG glioma cells. Although PDGF-B stimulated tyrosine phosphorylation of PDGF-Rbeta in U87MG cells, treatment with recombinant PDGF-B or overexpression of PDGF-B in U87MG cells had no effect on their proliferation. However, an increase of secreted PDGF-B in conditioned media of U87MG/PDGF-B cells promoted migration of endothelial cells expressing PDGF-R beta, whereas conditioned media from U87MG cells did not increase the cell migration. In mice, overexpression of PDGF-B in U87MG cells enhanced intracranial glioma formation by stimulating vascular endothelial growth factor (VEGF) expression in neovessels and by attracting vessel-associated pericytes. When PDGF-B and VEGF were overexpressed simultaneously by U87MG tumors, there was a marked increase of capillary-associated pericytes as seen in U87MG/VEGF(165)/PDGF-B gliomas. As a result of pericyte recruitment, vessels induced by VEGF in tumor vicinity migrated into the central regions of these tumors. These data suggest that PDGF-B is a paracrine factor in U87MG gliomas, and that PDGF-B enhances glioma angiogenesis, at least in part, by stimulating VEGF expression in tumor endothelia and by recruiting pericytes to neovessels. PMID- 12651602 TI - Molecular genetic evidence for monoclonal origin of bilateral ovarian serous borderline tumors. AB - Patients with serous borderline tumors of the ovary often present with multiple tumors at different sites in the abdominal cavity. Whether different foci of ovarian serous borderline tumors are monoclonal in origin, arising as a consequence of spread from a single ovarian site, or whether such deposits are polyclonal and explained by independent molecular genetic alterations on the background of a field defect, is unknown. So far, only X-chromosome inactivation studies were performed to study this issue. We used a genome-wide allelotyping to assess clonality in 47 metachronous and/or synchronous multifocal tumors from 22 patients, using 59 microsatellite markers. Loss of heterozygosity (LOH) was observed in only 34 of 1969 informative markers in 9 of 22 serous borderline cases studied. Of these cases, 7 showed concordant LOH for at least one polymorphic marker in more than one tumor site. Flanking microsatellite markers enabled identification of identical chromosomal breakpoints in 6 of 7 cases. The LOH results strongly favor a common origin indicated by a likelihood ratio (possibility common origin/possibility independent origin) ranging from 39 to 14,163. Strong additional evidence for monoclonality is provided by the finding of identical microsatellite alterations in all three-tumor sites in one case. PMID- 12651603 TI - Oral prion infection requires normal numbers of Peyer's patches but not of enteric lymphocytes. AB - Prion pathogenesis following oral exposure is thought to involve gut-associated lymphatic tissue, which includes Peyer's patches (PPs) and M cells. Recruitment of activated B lymphocytes to PPs requires alpha(4)beta(7) integrin; PPs of beta 7(-/-) mice are normal in number but are atrophic and almost entirely devoid of B cells. Here we report that minimal infectious dose and disease incubation after oral exposure to logarithmic dilutions of prion inoculum were similar in beta 7( /-) and wild-type mice, and PPs of both beta 7(-/-) and wild-type mice contained 3-4 log LD(50)/g prion infectivity > or =125 days after challenge. Despite marked reduction of B cells, M cells were present in beta 7(-/-) mice. In contrast, mice deficient in both tumor necrosis factor and lymphotoxin-alpha (TNF alpha(-/-) x LT alpha(-/-)) or in lymphocytes (RAG-1(-/-), mu MT), in which numbers of PPs are reduced in number, were highly resistant to oral challenge, and their intestines were virtually devoid of prion infectivity at all times after challenge. Therefore, lymphoreticular requirements for enteric and for intraperitoneal uptake of prions differ from each other. Although susceptibility to prion infection following oral challenge correlates with the number of PPs, it is remarkably independent of the number of PP-associated lymphocytes. PMID- 12651604 TI - API2-MALT1 fusion defines a distinctive clinicopathologic subtype in pulmonary extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue. AB - t(11;18)(q21;q21) is associated with mucosa-associated lymphoid tissue (MALT) type lymphoma and results in API2-MALT1 fusion. However, its clinicopathologic significance remains unclarified. API2-MALT1 fusion is detected most frequently in MALT lymphomas primarily involving the lung. We therefore screened 51 cases of pulmonary MALT lymphoma for API2-MALT1 fusion, and studied its relationship with clinicopathologic factors including the immunohistochemical expression of BCL10, another MALT lymphoma-associated molecule. The API2-MALT1 fusion transcript was detected in 21 of 51 (41%) cases, and was correlated with the absence of any underlying autoimmune disease, and with a normal serum lactate dehydrogenase, a "typical" histology without marked plasmacytic differentiation or an increased number of large cells, and aberrant nuclear BCL10 expression. However, its prognostic impact was not identified in the limited follow-up (6 to 187 months, median 27). These data suggest that the API2-MALT1 fusion may be a causative gene abnormality unrelated to autoimmune disease. In addition, this alteration may define a homogeneous MALT lymphoma subtype that is clinically more indolent and histologically more "typical." Aberrant nuclear BCL10 expression may have a possible role as a tool to screen for this API2-MALT1 fusion. A large-scale study with a long follow-up is necessary to establish the prognostic significance of API2-MALT1 fusion. PMID- 12651605 TI - RAGE drives the development of glomerulosclerosis and implicates podocyte activation in the pathogenesis of diabetic nephropathy. AB - Diabetic nephropathy ensues from events involving earliest changes in the glomeruli and podocytes, followed by accumulation of extracellular matrix in the mesangium. Postulated mechanisms include roles for vascular endothelial growth factor (VEGF), produced by podocytes and contributing to enhanced excretion of urinary albumin and recruitment/activation of inflammatory cells, and transforming growth factor-beta (TGF-beta), elicited largely from mesangial cells and driving production of extracellular matrix. RAGE, a receptor for advanced glycation endproducts (AGEs) and S100/calgranulins, displays enhanced expression in podocytes of genetically diabetic db/db mice by age 13 weeks. RAGE-bearing podocytes express high levels of VEGF by this time, in parallel with enhanced recruitment of mononuclear phagocytes to the glomeruli; events prevented by blockade of RAGE. By age 27 weeks, soluble RAGE-treated db/db mice displayed diminished albuminuria and glomerulosclerosis, and improved renal function. Diabetic homozygous RAGE null mice failed to develop significantly increased mesangial matrix expansion or thickening of the glomerular basement membrane. We propose that activation of RAGE contributes to expression of VEGF and enhanced attraction/activation of inflammatory cells in the diabetic glomerulus, thereby setting the stage for mesangial activation and TGF-beta production; processes which converge to cause albuminuria and glomerulosclerosis. PMID- 12651606 TI - Human urinary bladder transitional cell carcinomas acquire the functional Fas ligand during tumor progression. AB - The interaction between FasL on tumor cells and Fas on lymphocytes may represent a tumor immune escape mechanism. We explored FasL expression and function in human urinary bladder transitional cell carcinomas (TCCs). FasL expression was observed in situ in 45% of TCCs (n = 45) and was absent in normal urothelium (n = 20). A correlation existed between FasL expression and high tumor grade (0% in G1, 14% in G2, and 75% in G3; P < 0.0001) and stage (13% in superficial Ta-T1 versus 81% in invasive T2-T4; P < 0.0001). FasL function was shown by the ability of two FasL-positive primary culture TCC cell lines (established from two FasL positive invasive TCCs) to induce Fas-mediated killing not only of conventional Fas-sensitive targets (such as Jurkat cells or phytohemagglutinin-lymphoblasts), but also of autologous T lymphocytes generated in a mixed lymphocyte tumor-cell culture. In addition, an association between FasL expression by TCC cells and activated caspase-8, -9, and -3 expression by interferon-gamma-producing CD8 positive tumor-infiltrating lymphocytes was observed in situ. Our results show a functional expression of TCC-expressed FasL that correlates with tumor progression. These results suggest that TCC-expressed FasL may induce apoptosis of anti-tumor T lymphocytes in vivo, providing new insights on the mechanisms involved in bladder TCC progression. PMID- 12651608 TI - Role of CD31/platelet endothelial cell adhesion molecule-1 expression in in vitro and in vivo growth and differentiation of human breast cancer cells. AB - Breast ductal carcinoma in situ is an intraductal proliferation of malignant epithelial cells that diffuse within the ductal system without stromal invasion. Our finding that a subset of these tumors express CD31/platelet endothelial cell adhesion molecule-1 suggests that breast cancer represents an informative model for studying the involvement of the molecule in the morphogenesis, differentiation, and diffusion of this disease. Transfection of CD31 in MDA-MB 231 cells caused reduction in growth, loss of CD44, and acquisition of a ductal morphology. The same effects were maintained in vivo, in which CD31(+) tumors grew with in situ-like aspects, papillary differentiation, and a secretory phenotype. CD44 was down-modulated, with the CD31(+) cells blocked in the G(1) phase. The morphology was highly similar to what was observed in some human CD31(+) ductal carcinomas in situ. MDA-MB-231 mock cells grew in solid sheets, lacking stromal material, and displaying high levels of CD44 and proliferation. CD31(+) cells acquired motility characteristics in in vitro assays, a finding confirmed in vivo by the diffusion of human tumor cells throughout the normal ducts residual in the murine mammary gland. In conclusion, CD31 expression reverts the undifferentiated morphology and aggressive behavior of MDA-MB-231 cells, indicating its active role in the morphogenesis of breast ductal in situ carcinomas. PMID- 12651607 TI - Exploration of global gene expression patterns in pancreatic adenocarcinoma using cDNA microarrays. AB - Pancreatic cancer is the fifth leading cause of cancer death in the United States. We used cDNA microarrays to analyze global gene expression patterns in 14 pancreatic cancer cell lines, 17 resected infiltrating pancreatic cancer tissues, and 5 samples of normal pancreas to identify genes that are differentially expressed in pancreatic cancer. We found more than 400 cDNAs corresponding to genes that were differentially expressed in the pancreatic cancer tissues and cell lines as compared to normal pancreas. These genes that tended to be expressed at higher levels in pancreatic cancers were associated with a variety of processes, including cell-cell and cell-matrix interactions, cytoskeletal remodeling, proteolytic activity, and Ca(++) homeostasis. Two prominent clusters of genes were related to the high rates of cellular proliferation in pancreatic cancer cell lines and the host desmoplastic response in the resected pancreatic cancer tissues. Of 149 genes identified as more highly expressed in the pancreatic cancers compared with normal pancreas, 103 genes have not been previously reported in association with pancreatic cancer. The expression patterns of 14 of these highly expressed genes were validated by either immunohistochemistry or reverse transcriptase-polymerase chain reaction as being expressed in pancreatic cancer. The overexpression of one gene in particular, 14 3-3 sigma, was found to be associated with aberrant hypomethylation in the majority of pancreatic cancers analyzed. The genes and expressed sequence tags presented in this study provide clues to the pathobiology of pancreatic cancer and implicate a large number of potentially new molecular markers for the detection and treatment of pancreatic cancer. PMID- 12651609 TI - Hepatic artery and portal vein remodeling in rat liver: vascular response to selective cholangiocyte proliferation. AB - Three-dimensional reconstruction of the biliary tree, hepatic artery, and portal vein in normal rats and rats fed alpha-naphthylisothiocyanate (ANIT), a compound that causes selective proliferation of epithelial cells (ie, cholangiocytes) that line the bile ducts, was performed. All hepatic structures in ANIT-fed rats branched 1.5 times more often than in normal rats, reflecting an increased number of segments, whereas the length of the biliary tree, hepatic artery, and portal vein remain unchanged. The length of the proximal vessel segments was uniform in both groups of rats whereas the length of distal segments decreased twofold in ANIT-fed rats, suggesting that small vessels preferentially undergo proliferation. In contrast, the length of all bile duct segments decreased twofold, suggesting that ANIT induced proliferation of all compartments of the biliary tree. The total volume of the biliary tree, hepatic artery, and portal vein was increased 18, 4, and 3 times, respectively, after ANIT feeding. The diameters of the bile ducts (range, 20 to 259 microm) and arterial (range, 21 to 276 microm) segments in ANIT-fed rats did not differ from normal rats (range, 21 to 245 microm and 20 to 265 microm, respectively). In contrast, the diameters of proximal venous segments in ANIT-fed rats were significantly less (316 +/- 68 micro m versus 488 +/- 89 micro m, P < 0.001). The data suggest that after experimentally induced cholangiocyte proliferation, the hepatic artery and portal vein also undergo marked proliferation, presumably to support the increased nutritional and functional demands of the proliferated bile ducts. The molecular mechanisms of these vascular changes remain to be determined. PMID- 12651610 TI - Cathepsin D specifically cleaves the chemokines macrophage inflammatory protein-1 alpha, macrophage inflammatory protein-1 beta, and SLC that are expressed in human breast cancer. AB - Cathepsin D (Cath-D) expression in human primary breast cancer has been associated with a poor prognosis. In search of a better understanding of the Cath D substrates possibly involved in cancer invasiveness and metastasis, we investigated the potential interactions between this protease and chemokines. Here we report that purified Cath-D, as well as culture supernatants from the human breast carcinoma cell lines MCF-7 and T47D, selectively degrade macrophage inflammatory protein (MIP)-1 alpha (CCL3), MIP-1 beta (CCL4), and SLC (CCL21). Proteolysis was totally blocked by the protease inhibitor pepstatin A, and specificity of Cath-D cleavage was demonstrated using a large chemokine panel. Whereas MIP-1 alpha and MIP-1 beta degradation was rapid and complete, cleavage of SLC was slow and not complete. Mass spectrometry analysis showed that Cath-D cleaves the Leu(58) to Trp(59) bond of SLC producing two functionally inactive fragments. Analysis of Cath-D proteolysis of a series of monocyte chemoattractant protein-3/MIP-1 beta hybrids indicated that processing of MIP-1 beta might start by cleaving off amino acids located in the C-terminal domain. In situ hybridization studies revealed MIP-1 alpha, MIP-1 beta, and Cath-D gene expression mainly in the stromal compartment of breast cancers whereas SLC transcripts were found in endothelial cells of capillaries and venules within the neoplastic tissues. Cath-D production in the breast carcinoma cell lines MCF-7 and T47D, as assessed by enzyme-linked immunosorbent assay of culture supernatants and cell lysates, was not affected by stimulation with chemokines such as interleukin-8 (CXCL8), SDF-1 (CXCL12), and SLC. These data suggest that inactivation of chemokines by Cath-D possibly influences regulatory mechanisms in the tumoral extracellular microenvironment that in turn may affect the generation of the antitumoral immune response, the migration of cancer cells, or both processes. PMID- 12651611 TI - Modulation of hepatic granulomatous responses by transgene expression of DAP12 or TREM-1-Ig molecules. AB - DAP12 (also known as KARAP) is a novel ITAM-bearing transmembrane adapter molecule that is expressed on the cell surface of natural killer cells, monocytes, dendritic cells, and macrophages. Several myeloid cell-specific DAP12 associating receptors, such as TREM receptor family, SIRP-beta1, and MDL-1 have been identified. The in vivo function of DAP12 and its associating molecules in inflammation has remained primarily unknown. To investigate DAP12 signaling during chronic inflammation, we constructed two adenoviral gene transfer vectors to express FLAG/DAP12 (Ad-FDAP12) and the extracellular domain of mouse TREM-1 and the Fc portion of human IgG1 (Ad-TREM-1 Ig), respectively, and observed their modulatory activities in a mouse model of hepatic granulomatous inflammation elicited by zymosan A. Mice were injected with zymosan A intravenously and 24 hours after zymosan A, they were injected with Ad-FDAP12 or Ad-TREM-1 Ig. Zymosan A-induced hepatic granuloma formation peaked at day 7 and markedly declined by day 10. Although adenoviral-mediated DAP12 gene transfer did not enhance granuloma formation by day 7, it sustained and enhanced granuloma formation beyond day 7. However, an anti-FLAG monoclonal antibody used to potentiate the signaling of adenoviral-derived DAP12, enhanced granuloma formation at day 7. In sharp contrast to the effect by Ad-FDAP12, transgene expression in the liver of soluble form of extracellular domain of TREM-1 as an antagonist of DAP12 signaling, remarkably inhibited zymosan A-induced granuloma formation at all time points examined. Our findings thus suggest that both DAP12 and TREM-1 are involved in the development of granulomatous responses in the liver. PMID- 12651612 TI - Malgun (clear) cell change in Helicobacter pylori gastritis reflects epithelial genomic damage and repair. AB - Cancers may develop in the background of genomic instability with accumulated mutations. Helicobacter pylori gastritis is characterized by acute foveolitis of the proliferative zone, which is found in any stage of the gastritis as long as the infection persists. Because acute foveolitis targets specifically the proliferative zone of pits, the proliferating epithelial cells are under severe and persistent mutagenic pressure. In H. pylori gastritis, a characteristic morphological change of epithelial cells, the malgun (clear) cell change is frequently present in association with acute foveolitis. Malgun cells have enlarged euchromatic nuclei and abundant cytoplasm. The expression of proliferating cell nuclear antigen and cytokeratin 8 are typically up-regulated in them indicating that they are mitotically and metabolically active. Here, we report evidence for DNA damage and repair in malgun cells. Significant double strand DNA breaks were shown by the consistent terminal dUTP nick-end labeling in the nuclei of malgun cells. Proteins related to DNA damage and repair, such as Ku, poly(ADP-ribosyl) polymerase, OGG1, and MSH2 were selectively up-regulated in malgun cells. Inducible nitric oxide synthase was also up-regulated. There were occasional bcl2- and p53-expressing cells suggesting that further steps of carcinogenesis took place at the single cell level. Our results suggest that the malgun cell change represents a characteristic morphological sign of cellular genomic damage and repair, and may be implicated in an early stage of carcinogenesis. It is suggested that acute foveolitis of the proliferative zone is a major pathogenetic step of gastric carcinogenesis in H. pylori gastritis. PMID- 12651613 TI - Advanced glycation end products and receptor for advanced glycation end products in AA amyloidosis. AB - Advanced glycation end products (AGEs) may be involved in either amyloidogenesis or complications related to amyloid. We hypothesized that AGEs may influence the pathogenesis of AA amyloidosis, and investigated the spatial and temporal relationship between AGEs, carboxy methyl lysine (CML), the AGE receptor (RAGE), and AA amyloid in humans and mice. Specimens from patients with AL and ATTR amyloidosis served as a control. Using immunohistochemistry, AGEs, CML, and RAGE were found within amyloid deposits, more commonly in AA amyloid than in AL amyloid and not in ATTR amyloid. Western blotting showed that multiple proteins (between 12 and >60 kd) are modified, but not the AA amyloid fibril protein itself. In the murine model of AA amyloidosis, we found a marked interindividual variability with respect to local and systemic CML levels, as well as to splenic RAGE transcription. Serum levels of CML correlated with the duration of the inflammatory response but not with amounts of splenic RAGE mRNA. Other as yet unidentified variables, especially of the heterogeneous group of AGEs, probably modulate transcription of RAGE and influence amyloidogenesis. CML serum levels, in turn, may prove useful in predicting patients at risk. PMID- 12651614 TI - Expression and regulation of Toll-like receptor 2 in rheumatoid arthritis synovium. AB - Toll-like receptors (TLRs) are involved in mediating cell activation on stimulation with microbial constituents. We investigated the role for TLRs in synovial fibroblast (SF) activation in rheumatoid arthritis (RA). We analyzed whether stimulation with interleukin-1 beta and tumor necrosis factor-alpha, cytokines present in RA synovium, influences expression of TLR genes in SFs. The effects were compared with those of treatment with lipopolysaccharide and a synthetic lipopeptide (sBLP). Gene expression was examined using quantitative polymerase chain reaction. TLR2-mediated cell activation was investigated by electromobility shift assay for nuclear factor-kappa B. To localize TLR2 expression in joint tissue sections of RA patients were stained using in situ hybridization. Expression of TLR2 in RA SFs was increased after treatment with interleukin-1 beta, tumor necrosis factor-alpha, lipopolysaccharide, and sBLP. Nuclear factor-kappa B translocation in SFs was triggered by TLR2-mediated cell stimulation. Synovial tissues from RA joints expressed TLR2 predominantly at sites of attachment and invasion into cartilage and bone. The observed elevated expression of TLR2 in RA SFs could be a consequence of direct exposure to microbial compounds or of the presence of inflammatory mediators in the joint. TLR-associated signaling pathways may contribute to the pathogenesis of RA, either by initiating or perpetuating activation of SFs. PMID- 12651615 TI - Fibronectin-alpha 4 beta 1 integrin-mediated blockade protects genetically fat Zucker rat livers from ischemia/reperfusion injury. AB - We tested a hypothesis that interactions between fibronectin (FN), the major extracellular matrix component, and its integrin alpha 4 beta 1 receptor is important in the development of ischemia/reperfusion injury of steatotic liver transplants. We examined the effect of connecting segment-1 (CS1) peptide facilitated blockade of FN-alpha 4 beta 1 interaction in a well-established steatotic rat liver model of ex vivo cold ischemia followed by iso transplantation. In this model, CS1 peptides were administered through the portal vein of steatotic Zucker rat livers before and after cold ischemic storage. Lean Zucker recipients of fatty liver transplants received an additional 3-day course of CS1 peptides after transplant. CS1 peptide therapy significantly inhibited the recruitment of T lymphocytes, neutrophil activation/infiltration, and repressed the expression of proinflammatory tumor necrosis factor-alpha and interferon gamma. Moreover, it resulted in selective inhibition of inducible nitric oxide synthase expression, peroxynitrite formation, and hepatic necrosis. Importantly, CS1 peptide therapy improved function/histological preservation of steatotic liver grafts, and extended their 14-day survival in lean recipients from 40% in untreated to 100% in CS1-treated OLTs. Thus, CS1 peptide-mediated blockade of FN alpha 4 beta 1 interaction protects against severe ischemia/reperfusion injury experienced otherwise by steatotic OLTs. These novel findings document the potential of targeting FN-alpha 4 beta 1 in vivo interaction to increase the transplant donor pool through modulation of marginal steatotic livers. PMID- 12651616 TI - Disruption of the caveolin-1 gene impairs renal calcium reabsorption and leads to hypercalciuria and urolithiasis. AB - Using LoxP/Cre technology, we generated a knockout mouse homozygous for a null mutation in exon 2 of Cav1. In male Cav1-/- animals, we observed a dramatic increase in the incidence of urinary calcium stone formation. In 5-month-old male mice, the incidence of early urinary calculi was 67% in Cav1-/- mice compared to 19% in Cav1+/+ animals. Frank stone formation was observed in 13% of Cav1-/- males but was not seen in Cav1+/+ mice. Urine calcium concentration was significantly higher in Cav1-/- male mice compared to Cav1+/+ mice. In Cav1-/- mice, distal convoluted tubule cells were completely devoid of Cav1 and the localization of plasma membrane calcium ATPase was disrupted. Functional studies confirmed that active calcium absorption was significantly reduced in Cav1-/- compared to Cav1+/+ male mice. These results demonstrate that disruption of the Cav1 gene promotes the progressive steps required for urinary calcium stone formation and establish a new mouse model for urinary stone disease. PMID- 12651617 TI - Potential interaction between CCR1 and its ligand, CCL3, induced by endogenously produced interleukin-1 in human hepatomas. AB - Hepatoma cell lines can produce a massive amount of chemokines in response to various stimuli including hepatitis viruses and their products. However, it remains elusive on the types of chemokine receptor(s) expressed in the hepatoma tissues and its roles in hepatoma development. To clarify these points, we examined the chemokine receptor expression in six human hepatoma cell lines. All of the hepatoma cell lines constitutively and exclusively expressed CCR1 mRNA and its protein on their cell surface. CCR1 expression was also detected on hepatoma cells and to a lesser degree, on endothelial cells in hepatoma tissues but not in normal liver tissues. Furthermore, CCL3 expression was detected in hepatoma cells, endothelial cells, and to a lesser degree, fibroblast-like cells in hepatoma tissue, whereas only occasional vascular endothelial cells and inflammatory cells in normal liver tissues were weakly positive for CCL3. Moreover, the forskolin-mediated increases in intracellular cAMP concentrations were inhibited by the ligands for CCR1, CCL3, CCL4, and CCL5, suggesting that the expressed CCR1 was functional. Four hepatoma cell lines produced CCL3 only in response to interleukin (IL)-1 alpha and IL-1 beta. Finally, IL-1 alpha and IL-1 beta were detected abundantly in hepatoma tissues but not in normal liver tissues. Thus, IL-1 may enhance the local production of CCL3, which may interact with CCR1 expressed on hepatoma cells, in an autocrine and/or paracrine manner. PMID- 12651619 TI - Urinary outflow obstruction increases apoptosis and deregulates Bcl-2 and Bax expression in the fetal ovine bladder. AB - During organogenesis, net growth of tissues is determined by a balance between proliferation, hypertrophy, and apoptotic death. Human fetal bladder outflow obstruction is a major cause of end-stage renal failure in children and is associated with complex pathology in the kidney and lower urinary tract. Experimental manipulation of the fetal sheep urinary tract has proved informative in understanding the pathobiology of congenital obstructive uropathy. In this study we used an ovine model of fetal bladder outflow obstruction to examine effects on apoptotic cell death in the developing urinary bladder. While 30 days of obstruction in utero between 75 and 105 days gestation resulted in overall growth of the fetal bladder as assessed by weight, protein, and DNA measurements, we found that apoptosis, as assessed by in situ end-labeling, was up-regulated in fetal bladder detrusor muscle and lamina propria cells and that this was accompanied by a down-regulation of the anti-death protein Bcl-2 and an up regulation of the pro-death protein Bax. Moreover, activated caspase-3, an effector of apoptotic death, was increased in obstructed bladders. This is the first study to define altered death in an experimental fetal model of bladder dysmorphogenesis. We speculate that enhanced apoptosis in detrusor smooth muscle cells is part of a remodeling response during compensatory hyperplasia and hypertrophy. Conversely, in the lamina propria, an imbalance between death and proliferation leads to a relative depletion of cells. PMID- 12651618 TI - Axonal injury heralds virus-induced demyelination. AB - Axonal pathology has been highlighted as a cause of neurological disability in multiple sclerosis. The Daniels (DA) strain of Theiler's murine encephalomyelitis virus infects the gray matter of the central nervous system of mice during the acute phase and persistently infects the white matter of the spinal cord during the chronic phase, leading to demyelination. This experimental infection has been used as an animal model for multiple sclerosis. The GDVII strain causes an acute fatal polioencephalomyelitis without demyelination. Injured axons were detected in normal appearing white matter at 1 week after infection with DA virus by immunohistochemistry using antibodies specific for neurofilament protein. The number of damaged axons increased throughout time. By 2 and 3 weeks after infection, injured axons were accompanied by parenchymal infiltration of Ricinus communis agglutinin I(+) microglia/macrophages, but never associated with perivascular T-cell infiltration or obvious demyelination until the chronic phase. GDVII virus infection resulted in severe axonal injury in normal appearing white matter at 1 week after infection, without the presence of macrophages, T cells, or viral antigen-positive cells. The distribution of axonal injury observed during the early phase corresponded to regions where subsequent demyelination occurs during the chronic phase. The results suggest that axonal injury might herald or trigger demyelination. PMID- 12651620 TI - Dominant-negative hypoxia-inducible factor-1 alpha reduces tumorigenicity of pancreatic cancer cells through the suppression of glucose metabolism. AB - In the tumor cells exposed to hypoxia, hypoxia-inducible factor-1 (HIF-1) mediated adaptation responses such as angiogenesis and anaerobic metabolism are induced for their survival. We have recently reported that the constitutive expression of HIF-1 alpha renders pancreatic cancer cells resistant to apoptosis induced by hypoxia and glucose deprivation. We then established dominant-negative HIF-1 alpha (dnHIF-1 alpha) transfectants and examined their susceptibility to apoptosis and growth inhibition induced by hypoxia and glucose deprivation in vitro and their tumorigenicity in SCID mice. We further examined the expressions of aldolase A and Glut-1 in vitro and Glut-1 expression and glucose uptake in the tumor tissues and microvessel counts in the tumor tissues. As a result, dnHIF-1 alpha rendered the pancreatic cancer cells sensitive to apoptosis and growth inhibition induced by hypoxia and glucose deprivation. Also it abrogated the enhanced expression of Glut-1 and aldolase A mRNAs under hypoxia and reduced the expression of Glut-1 and the glucose uptake in the tumor tissues and consequently in vivo tumorigenicity. We found no significant difference in the microvessel counts among the tumor tissues. From these results, we suggest that the disruption of the HIF-1 pathway might be effective in the treatment of pancreatic cancers. PMID- 12651621 TI - Transglutaminase type II plays a protective role in hepatic injury. AB - The up-regulation of "tissue" transglutaminase (TG2) gene has been shown to occur in various pathologies and can lead to severe liver injury; however, its role in the onset of liver damage has not yet been clarified. To address this issue, we have used two experimental settings: carbon tetrachloride (CCl(4))-induced liver injury in wild-type and TG2 knockout mice; and liver biopsies obtained from a large cohort of hepatitis C virus (HCV)-infected patients. Mice lacking TG2 failed to clear the hepatic necrotic tissue formed in response to prolonged CCl(4) exposure (5 weeks) and 60% of them died before the end of the treatment. By contrast, wild-type mice were able to recover after the toxic insult. CCl(4) treated TG2 null mice showed a derangement of the hepatic lobular architecture and a progressive accumulation of extracellular matrix (ECM) components and inflammatory cells which were not observed in the liver of control animals. Consistent with this protective role, we observed that TG2 levels were much higher (up to 15-fold) during the initial stages of liver fibrosis in HCV infected individuals (METAVIR = F2) compared with uninfected controls, in which the enzyme protein localized in the hepatocytes facing the periportal infiltrate. By contrast, the enzyme levels decreased in the advanced stages (METAVIR = F3 and F4) and their localization was limited to the ECM. Our data demonstrate that TG2 plays a protective role in the liver injury by favoring tissue stability and repair. PMID- 12651622 TI - Telomere shortening and cellular senescence in a model of chronic renal allograft rejection. AB - Cellular senescence has been suggested to play a role in the deterioration of renal graft function and has been linked to telomere shortening. We have investigated markers of cellular senescence in the F344 to LEW rat model of chronic renal transplant rejection. Syngeneic and LEW to F344 transplants were used as controls. Substantial telomere shortening was observed in all transplants, including allogeneic and syngeneic grafts from day 7 post-transplant onwards. Ischemia of native F344 kidneys was already sufficient to induce telomere shortening. It is known that shortened telomeres can activate cell cycle regulators, such as p21 and p16. Accordingly, all cases showed a transient p21 increase, with a maximum at day 7 and a sustained expression of p16. Importantly, senescence-associated beta-galactosidase staining, a cytological marker for senescence, was only observed in tubular epithelial cells of chronically rejecting F344 allografts from day 30 post-transplantation onwards. Long-term surviving LEW allografts or syngeneic F344 grafts were negative for senescence associated beta-galactosidase. In conclusion, ischemia during transplantation results in telomere shortening and subsequent activation of p21 and p16, whereas senescence-associated beta-galactosidase staining is only present in chronically rejecting kidney grafts. PMID- 12651623 TI - The human Fn14 receptor gene is up-regulated in migrating glioma cells in vitro and overexpressed in advanced glial tumors. AB - Glioblastoma multiforme comprises the majority of human brain tumors. Patients with glioblastoma multiforme have poor survival rates, with an average life expectancy of <1 year. To assess possible mechanisms and to potentially target invasive glioma cells, we previously measured the gene expression profiles of glioma cells under migration-activated or passive states. One of the genes identified was Fn14, which encodes a cell surface receptor for the tumor necrosis factor superfamily member named TWEAK. In this study, we show that Fn14 gene expression is induced in migration-activated glioma cells in vitro and significantly increases according to tumor grade in vivo (P < 0.01), with highest levels in glioblastoma tissue specimens. The in situ expression pattern of Fn14 mRNA and protein was confined to primary glioma cells and the vascular endothelium, with no detection in adjacent normal brain. Conversely, TWEAK mRNA levels are low in glioblastoma samples relative to normal brain tissue. In addition, activation of the Fn14 receptor by addition of recombinant TWEAK resulted in increased glioma cell migration in vitro. These results suggest a positive role for TWEAK and Fn14 in glioma progression and indicate that Fn14 gene expression may serve as a marker for invasive glioma cells. PMID- 12651624 TI - Expression of E-cadherin and other paracellular junction genes is decreased in iron-loaded hepatocytes. AB - Iron overload in the liver may occur in the clinical conditions hemochromatosis and transfusion-dependent thalassemia or by long-term consumption of large amounts of dietary iron. As iron concentrations increase in the liver, cirrhosis develops, and subsequently the normal architecture of the liver deteriorates. The underlying mechanisms whereby iron loading of hepatocytes leads to the pathology of the liver are not understood. Similarly, a direct relationship between the expression levels of paracellular junction genes and altered hepatocellular physiology has been reported; however, no relationship has been identified between iron loading and the expression of paracellular junction genes. Here, we report that the expression of numerous paracellular junction genes was decreased in iron-loaded hepatocytes, leading to increased cellular permeability, increased baculovirus-mediated gene transfer, and decreased gap junction communication. Iron loading of hepatocytes resulted in decreased E-cadherin promoter activity and subsequently decreased E-cadherin mRNA and protein expression. The data presented in this study describe a clear relationship between iron overload and decreased expression of paracellular junction genes in hepatic cells of rat and human origin. PMID- 12651625 TI - Resistance to glomerulosclerosis in B6 mice disappears after menopause. AB - The frequency of chronic renal failure increases with age, especially in women after menopause. Glomerulosclerosis is a common cause of chronic renal failure in aging. We reported that pre-menopausal female C57BL6 (B6) mice are resistant to glomerulosclerosis, irrespective of the type of injury. However, we now show that B6 mice develop progressive glomerulosclerosis after menopause. Glomerular lesions, first recognized in 18-month-old mice, consisted of hypertrophy, vascular pole sclerosis, and mesangial cell proliferation. Diffuse but moderate mesangial sclerosis and more marked hypertrophy were present at 22 months. At 28 to 30 months the glomerulosclerosis was diffuse and increased levels of type I and type IV collagen and transforming growth factor-beta 1 mRNA were present. Urine albumin excretion was significantly increased in 30-month-old mice. Mesangial cells isolated from 28-month-old mice retained their sclerotic phenotype in vitro. Comparison of the effects of uninephrectomy (Nx) in 20-month old and 2.5-month-old mice revealed a 1.7-fold increase in urine albumin excretion, accelerated glomerulosclerosis, and renal function insufficiency in 20 month-old Nx mice, but not in 2.5-month-old Nx mice. Glycemic levels, glucose, insulin tolerance, and blood pressure were normal at all ages. Thus, B6 mice model the increased frequency of chronic renal failure in postmenopausal women and provide a model for studying the mechanism(s) of glomerulosclerosis in aging women. PMID- 12651626 TI - Deletion, mutation, and loss of expression of KLF6 in human prostate cancer. AB - Kruppel-like factors (KLFs) are a group of transcription factors that appear to be involved in different biological processes including carcinogenesis. In a recent study, KLF6 was reported as a tumor suppressor gene in prostate cancer because of its frequent loss of heterozygosity (LOH) and mutation as well as functional suppression of cell proliferation. Loss of chromosomal locus spanning KLF6 is relatively infrequent in other published studies of prostate cancer, however. To clarify the role of KLF6 in prostate cancers, particularly those that are high grade, we examined KLF6 for deletion, mutation, and loss of expression in 96 prostate cancer samples including 21 xenografts/cell lines. Loss of heterozygosity occurred in 4 (19%) of 21 xenografts/cell lines and 8 (28%) of 29 informative tumors. Fourteen of the 96 (15%) samples showed 15 somatic sequence changes in the KLF6 gene, including 7 that changed KLF6 peptide sequences, 4 that did not, and 4 that were located in untranslated regions. Expression levels of KLF6 were significantly lost in 4 of 20 (20%) xenografts/cell lines of prostate cancer, as detected by RT-PCR and Northern blot analysis. These findings indicate that significant genetic alterations of KLF6 occur in a minority of high-grade prostate cancers. PMID- 12651627 TI - Expression and regulation of tissue inhibitor of metalloproteinase-1 and matrix metalloproteinases by intestinal myofibroblasts in inflammatory bowel disease. AB - Intestinal fibrosis and strictures frequently occur in Crohn's disease but not ulcerative colitis. We have recently shown that, compared to myofibroblasts obtained from normal and ulcerative colitis tissue, myofibroblasts isolated from fibrotic Crohn's disease mucosal samples express significantly lower amounts of transforming growth factor (TGF)-beta 3, but the expression of TGF-beta 2 was significantly greater. We now report that in myofibroblast cultures established from fibrotic Crohn's disease mucosal samples there is significantly higher constitutive expression of tissue inhibitor of metalloproteinase (TIMP)-1 compared to similar cells isolated from normal or ulcerative colitis tissue. Myofibroblasts derived from normal mucosa and from mucosa affected by ulcerative colitis or Crohn's disease also expressed matrix metalloproteinase (MMP)-1, MMP 2, and MMP-3 but did not express MMP-9. Recombinant (r) TGF-beta 1 and rTGF-beta 2, but not rTGF-beta 3, induced expression of TIMP-1 in normal intestinal myofibroblasts. These studies illustrate a potential mechanism by which differential expression of isoforms of TGF-beta may lead to excessive deposition of extracellular matrix and stricture formation via TIMP-1-mediated inhibition of MMP activity. PMID- 12651628 TI - The relationship between hypomethylation and CpG island methylation in colorectal neoplasia. AB - Tumors are often characterized by an imbalance in cytosine methylation as manifested both by hypermethylation of CpG islands and by genome hypomethylation. These epigenetic changes were assessed in colorectal neoplasia to determine whether they arose through a common mechanism or indeed were distinct and unrelated phenomena. Fresh representative samples of adenomas, hyperplastic polyps, colorectal cancers, and normal mucosa were used in this study. Global methylation levels were measured by analyzing the methyl-accepting capacity of DNA. Methylation of p16, hMLH1, and MINT 1, 2, 12, and 31 were assessed by bisulfite polymerase chain reaction. Microsatellite status was determined by polymerase chain reaction using six markers and hMLH1 and proliferating cell nuclear antigen expression was assessed by immunohistochemistry. Normal colonic mucosa had a higher endogenous 5-methyl cytosine content than all proliferative lesions of the colon (P < 0.001). The extent of demethylation in hyperplastic polyps and adenomas was significantly related to its proliferative rate. Right sided hyperplastic polyps were more likely to be methylated than adenomas (odds ratio, 2.3; confidence interval, 1.1 to 4.6). There was no relationship between the level of global hypomethylation and hypermethylation. Some hyperplastic colorectal polyps have a propensity to develop dense CpG island methylation. Hypermethylation and hypomethylation contribute separately to the process of carcinogenesis. PMID- 12651629 TI - Functional significance of inducible nitric oxide synthase induction and protein nitration in the thermally injured cutaneous microvasculature. AB - Increased nitric oxide (NO) production after burn injury is well established. However, there is little information relating to the reactions that occur as a consequence of NO generation under such circumstances. We have investigated the synthesis and function of NO in a rat model of local cutaneous thermal injury. We show that NO levels are elevated from 3 hours after injury with a concomitant increase in protein nitration. A selective inducible nitric oxide synthase (iNOS) inhibitor (1400W) significantly attenuated NO synthesis, protein nitration, and neutrophil accumulation in this model, but had no effect on edema formation. The results also indicate that NO synthesis and protein nitration occurred independently of neutrophil accumulation because these parameters were unaffected by depletion of circulating neutrophils. 3-Chlorotyrosine, a marker of neutrophil/myeloperoxidase-mediated protein damage was significantly increased from 1 hour after burn. Our observations provide evidence for the involvement of reactive species in the inflammatory response after burn. The use of selective iNOS inhibitors may represent a novel approach for the management of human burn injuries. PMID- 12651630 TI - Switch moiety in agonist/antagonist dual effect of S19 ribosomal protein dimer on leukocyte chemotactic C5a receptor. AB - The cross-linked homodimer of S19 ribosomal protein (RP S19) induces monocyte predominant infiltration due to a dual effect on the C5a receptor in leukocyte chemotaxis, agonistic to monocytes and antagonistic to polymorphonuclear leukocytes (PMN) (H. Nishiura, Y. Shibuya, T. Yamamoto, Lab Invest 1998, 78:1615 1623). The agonistic ligand moiety was recently determined to be -Leu131-Asp132 Arg133- (Y. Shibuya et al, Am J Pathol 2001, 159:2293-2301). In this study we determined the moiety responsible for the antagonistic function. A C-terminal analogue peptide of RP S19, with 18 residues containing the agonistic ligand moiety as a part, reproduced the dual function in the leukocyte chemotaxis. A C5a analogue peptide attracted PMN as well as monocytes. When C-terminal 12 residues of RP S19 after the agonistic moiety, IAGQVAAANKKH, were connected to the C5a peptide, the chimeric peptide newly obtained the dual function, indicating that the C-terminal portion of RP S19 functions as a converter from the agonist to the antagonist. C-terminal truncation analyses indicated that the C-terminal His was not essential but the next Lys was necessary for the converter function. The homodimer of a mutant RP S19 that was truncated for the C-terminal 4 residues lost the monocyte selectivity in the leukocyte infiltration in vivo as in the case of the leukocyte chemotaxis in vitro. These results indicated that the conversion of the RP S19 dimer from agonist to antagonist of C5a receptor is attributed to the IAGQVAAANKK moiety between Ile134 and Lys144. PMID- 12651631 TI - Bovine thrombin and systemic autoimmunity. PMID- 12651632 TI - CFTR activation in human bronchial epithelial cells by novel benzoflavone and benzimidazolone compounds. AB - Activators of the CFTR Cl- channel may be useful for therapy of cystic fibrosis. Short-circuit current (Isc) measurements were done on human bronchial epithelial cells to characterize the best flavone and benzimidazolone CFTR activators identified by lead-based combinatorial synthesis and high-throughput screening. The 7,8-benzoflavone UCcf-029 was a potent activator of Cl- transport, with activating potency (<1 microM) being much better than other flavones, such as apigenin. The benzimidazolone UCcf-853 gave similar Isc but with lower potency (5 20 microM). In combination, the effect induced by maximal UCcf-029 and UCcf-029, UCcf-853, and apigenin increased strongly with increasing basal CFTR activity: for example, Kd for activation by UCcf-029 decreased from >5 to <0.4 microM with increasing basal Isc from approximately 4 microA/cm2 to approximately 12 microA/cm2. This dependence was confirmed in permeabilized Fischer rat thyroid cells stably expressing CFTR. Our results demonstrate efficacy of novel CFTR activators in bronchial epithelia and provide evidence that activating potency depends on basal CFTR activity. PMID- 12651633 TI - Deficiency in the c-Jun NH2-terminal kinase signaling pathway confers susceptibility to hyperoxic lung injury in mice. AB - Hyperoxia generates an oxidative stress in the mouse lung, which activates the major stress-inducible kinase pathways, including c-Jun NH2-terminal kinase (JNK). We examined the effect of Jnk1 gene deletion on in vivo responses to hyperoxia in mice. The survival of Jnk1-/- mice was reduced relative to wild-type mice after exposure to continuous hyperoxia. Jnk1-/- mice displayed higher protein concentration in bronchoalveolar lavage (BAL) fluid and increased expression of heme oxygenase-1, a stress-inducible gene, after 65 h of hyperoxia. Contrary to other markers of injury, the leukocyte count in BAL fluid of Jnk1-/- mice was markedly diminished relative to that of wild-type mice. The decrease in BAL leukocyte count was not associated with any decrease in lung myeloperoxidase activity at baseline or after hyperoxia treatment. Pretreatment with inhaled lipopolysaccharide increased BAL neutrophil content and extended hyperoxia survival time to a similar extent in Jnk1-/- and wild-type mice. Associated with increased mortality, Jnk1-/- mice had increased pulmonary epithelial cell apoptosis after exposure to hyperoxia compared with wild-type mice. These results indicate that JNK pathways participate in adaptive responses to hyperoxia in mice. PMID- 12651634 TI - Hyperchloremic acidosis: the classic example of strong ion acidosis. PMID- 12651635 TI - Single-dose tranexamic acid reduces postoperative bleeding after coronary surgery in patients treated with aspirin until surgery. AB - Tranexamic acid reduces postoperative bleeding after coronary artery bypass grafting. We evaluated the effects of a single dose of tranexamic acid given immediately before cardiopulmonary bypass (CPB) in patients treated with aspirin until the day before surgery. The study was a prospective, randomized, double blinded, placebo-controlled, parallel-group trial. Eighty patients were included and divided into two groups: one group received tranexamic acid 30 mg/kg, and one group received placebo (0.9% NaCl) as a bolus injection before CPB. Postoperative blood loss was recorded for 16 h. Transfusions of blood products were recorded for the whole hospital stay. Transfusions of packed red cells were given when the hematocrit value was less than 20% during CPB and less than 25% after surgery. The patients in the tranexamic acid group had significantly less postoperative bleeding compared with the patients in the placebo group (mean [SD]) (475 [274] mL versus 713 [243] mL; P < 0.001). An effective inhibition of fibrinolysis was found in patients receiving tranexamic acid. Tranexamic acid reduces postoperative bleeding in coronary artery bypass grafting patients treated with aspirin until the day before surgery. IMPLICATIONS: Continuation of aspirin medication until the day before coronary artery bypass grafting may increase postoperative bleeding. The administration of a single dose of tranexamic acid (30 mg/kg) immediately before cardiopulmonary bypass significantly reduced postoperative bleeding and inhibited fibrinolysis in these patients. PMID- 12651636 TI - Hemostasis and hemodilution: a quantitative mathematical guide for clinical practice. AB - Quantitative changes of hemostasis during hemodilution remain unclear. With the increasing popularity of artificial blood substitutes (ABS), which solely provide oxygen-transport capacity, this issue becomes even more complex. We developed a mathematical model to quantitatively analyze hemostasis during hemodilution and validated it by recalculating patient data. We calculated and compared maximal allowable blood losses (MABL) related to minimal acceptable hematocrit, platelet concentration, and plasma fibrinogen concentration. MABL is the maximal blood loss that can be tolerated without any additional blood products. The variable with the smallest MABL thus limits hemodilution foremost. Hemodilution included isovolemic replacement of blood loss with colloid or acute normovolemic hemodilution (ANH) followed by isovolemic replacement of blood loss with colloid and ABS. We also related our findings to preoperative patient data (n = 204). The decline in platelet concentrations rarely (<2% of all patients) limits hemodilution. By contrast, critical plasma fibrinogen (< or =100 mg/dL) concentrations can often (< or =20% of all patients) limit hemodilution if their initial concentrations are within the lower normal range (<300 mg/dL). These findings become more frequent if ANH is combined with ABS. Under those circumstances ANH blood products are solely required for stabilization of hemostasis, thereby defeating the original purpose of combining ANH with ABS. IMPLICATIONS: The causes of quantitative changes of hemostasis during hemodilution, as well as their clinical effect and relevance, remain unclear. Using a validated, realistic mathematical model, we demonstrate that hemostasis, especially plasma fibrinogen, can limit the extent of hemodilution. This phenomenon is particularly prominent when acute normovolemic hemodilution is combined with artificial blood substitutes. PMID- 12651637 TI - Hydroxyethyl starch (HES) 130/0.4 provides larger and faster increases in tissue oxygen tension in comparison with prehemodilution values than HES 70/0.5 or HES 200/0.5 in volunteers undergoing acute normovolemic hemodilution. AB - Stable hemodynamics and improved rheology are important effects of hemodilution with hydroxyethyl starch (HES) infusions. One clinical indicator of improved rheology is increased tissue oxygen tension (tpO(2)). In this prospective, randomized, double-blinded, crossover study, we examined the effects of acute normovolemic hemodilution with HES 130/0.4 on hemodynamics and skeletal muscle tpO(2) in comparison with conventional HES solutions. Twelve healthy volunteers were randomly enrolled in each group. At an interval of >8 days, volunteers donated 18% of their calculated blood volume within 30 min and randomly received 6% HES 130/0.4, 6% HES 70/0.5, or 6% HES 200/0.5 (crossover design) in a 1:1.2 ratio to their blood loss. Hemodynamic variables, tpO(2) in the quadriceps muscle, hematocrit, plasmatic HES concentrations, plasma viscosity, colloid osmotic pressures, and platelet aggregation were measured until 6 h after the infusion of HES. No differences were found among groups with respect to changes of hemodynamics, hematocrit, or platelet aggregation. With HES 200, colloid osmotic pressures and plasma viscosities were larger than after HES 70 (P < 0.05). HES 130 in comparison with HES 70 and 200 caused the fastest (30 min versus 90 min and 150 min after hemodilution; P < 0.05) and largest increase of tpO(2) in comparison to baseline (+93% versus +33% and 40%; P < 0.05). In healthy volunteers undergoing acute normovolemic hemodilution, the newly designed HES 130/0.4 showed a more pronounced and earlier increase of skeletal muscle tpO(2) in comparison with prehemodilution values than HES 70/0.5 or 200/0.5. IMPLICATIONS: The effects of three different hydroxyethyl starch (HES) solutions on hemodynamics, rheology, and skeletal muscle tissue tension after acute normovolemic hemodilution were examined in awake volunteers. With HES 130/0.4, increases of tissue oxygen tension in comparison to baseline were larger and more rapid than with HES 70/0.5 or HES 200/0.5. PMID- 12651638 TI - The effect of the interval between blood pressure determinations on the delay in the detection of changes: a computer simulation. AB - The frequency of automated noninvasive blood pressure (NIBP) measurements during routine anesthesia is a balance between potentially deleterious effects of frequent cycling and a delay in detecting changes caused by a long cycle time. A computer model generated systolic blood pressures that changed to a new, random value after a period of stability. We sampled these data at intervals between 1 and 10 min to simulate NIBP measurements. A separate algorithm, based on Trigg's Tracking Variable, indicated when a change had been detected. For each set of variables, the simulation was repeated 1000 times, and the average time to detect a change was recorded. The mean time to detect a change was 8.0 min with a 1-min cycle, 8.9 min with a 2-min cycle, 10.8 min with a 5-min cycle, and 13.0 min with a 10-min cycle. As the cycle time increased, the delay in detecting changes increased but only by approximately half the increase in the cycle time. The optimum variables for the trend detection algorithm also changed as the NIBP interval increased. Provided that abrupt changes in blood pressure are not anticipated, a 1- or 2-min cycle time for NIBP offers little advantage over a longer period. IMPLICATIONS: We used a computer model to study the effect of increasing noninvasive blood pressure (NIBP) sampling interval on the detection of blood pressure changes. The detection time increased only 50% of the increase in the sampling interval. This information may help optimize NIBP intervals in different circumstances. PMID- 12651640 TI - Hemostatic analysis of a patient undergoing off-pump coronary artery bypass surgery with argatroban anticoagulation. AB - IMPLICATIONS: This case describes the impact of argatroban and off-pump coronary revascularization on hemostasis as assessed by conventional hemostatic measures and Thrombelastography in a patient with heparin-induced thrombocytopenia. PMID- 12651639 TI - Sevoflurane exposure generates superoxide but leads to decreased superoxide during ischemia and reperfusion in isolated hearts. AB - Reactive oxygen species (ROS) are largely responsible for cardiac injury consequent to ischemia and reperfusion, but, paradoxically, there is evidence suggesting that anesthetics induce preconditioning (APC) by generating ROS. We hypothesized that sevoflurane generates the ROS superoxide (O(2)(.-)), that APC attenuates O(2)(.-) formation during ischemia, and that this attenuation is reversed by bracketing APC with the O(2)(.-) scavenger manganese (III) tetrakis (4-benzoic acid) porphyrin chloride (MnTBAP) or the putative mitochondrial adenosine triphosphate-sensitive potassium (mK(ATP)) channel blocker 5 hydroxydecanoate (5-HD). O(2)(.-) was measured continuously in guinea pig hearts by using dihydroethidium. Sevoflurane was administered alone (APC), with MnTBAP, or with 5-HD before 30 min of ischemia and 120 min of reperfusion. Control hearts underwent no pretreatment. Sevoflurane directly increased O(2)(.-); this was blocked by MnTBAP but not by 5-HD. O(2)(.-) increased during ischemia and during reperfusion. These increases in O(2)(.-) were attenuated in the APC group, but this was prevented by MnTBAP or 5-HD. We conclude that sevoflurane directly induces O(2)(.-) formation but that O(2)(.-) formation is decreased during subsequent ischemia and reperfusion. The former effect appears independent of mK(ATP) channels, but not the latter. Our study indicates that APC is initiated by ROS that in turn cause mK(ATP) channel opening. Although there appears to be a paradoxical role for ROS in triggering and mediating APC, a possible mechanism is offered. IMPLICATIONS: Reactive oxygen species (ROS) are implicated in triggering anesthetic preconditioning (APC). The ROS superoxide (O(2)(.-)) was measured continuously in guinea pig isolated hearts. Sevoflurane directly increased O(2)(. ) but led to attenuated O(2)(.-) formation during ischemia. This demonstrates triggering of APC by ROS and clarifies the mechanism of cardioprotection during ischemia. PMID- 12651641 TI - An unusual cause of seizures after cardiopulmonary bypass. AB - IMPLICATIONS: We present a case of a 34-yr-old woman who developed seizures after an open-heart procedure requiring cardiopulmonary bypass. The seizures were most likely caused by previously undiagnosed neurocysticercosis and may also have been contributed to by cardiopulmonary bypass. PMID- 12651643 TI - A liberalized fasting guideline for formula-fed infants does not increase average gastric fluid volume before elective surgery. AB - Recommended preoperative fasting intervals for infant formula vary from 4 to 8 h. We conducted a prospective, randomized, observer-blinded trial of 97 ASA physical status I and II infants scheduled for elective surgery to determine whether average gastric fluid volume (GFV) recovered from infants formula-fasted for 4 h (liberalized fast, Group L) differed from that recovered from infants allowed clear liquids up until 2 h, but fasted 8 h for formula and solids (traditional fast, Group T). In Group L, 31 of 39 subjects followed protocol and ingested formula 4-6 h before surgery. In Group T, 36 of 58 subjects followed protocol, taking clear liquids 2-5 h before the induction of anesthesia. Thirty subjects had prolonged fasts and were included only in a secondary intent-to-treat analysis. Respective mean age (5.7 +/- 2.3 versus 6.4 +/- 2.4 mo; range, 0.7-10.5 mo), weight (7.5 +/- 1.8 versus 7.5 +/- 1.1 kg), and volume of last feed (4.9 +/- 2.2 versus 4.0 +/- 2.3 oz.) did not vary between Groups L and T. GFV (L: 0.19 +/- 0.38 versus T: 0.16 +/- 0.30 mL/kg) and gastric fluid pH (L: 2.5 +/- 0.5 versus T: 2.9 +/- 1.3) did not vary. For all subjects, GFV (mL/kg) increased with age (Spearman correlation coefficient = +0.23, P = 0.03). Infant irritability and hunger and parent satisfaction were similar between groups. We conclude that average GFV after either a 4- to 6-h fast for infant formula or 2-h fast after clear liquids is small and not significantly different between groups. On the basis of these findings, clinicians may consider liberalizing formula feedings to 4 h before surgery in selected infants. IMPLICATIONS: Healthy infants aged < or =10.5 mo may drink formula up to 4 h before surgery without increasing gastric fluid volume compared with infants allowed clear liquids up to 2 h and formula 8 h before surgery. PMID- 12651642 TI - Transesophageal echocardiographic diagnosis of carbon dioxide embolism during minimally invasive saphenous vein harvesting and treatment with inhaled epoprostenol. AB - IMPLICATIONS: We describe a patient scheduled for coronary artery bypass who developed carbon dioxide (CO2) embolism with acute pulmonary hypertension during endoscopic saphenectomy. Transesophageal echocardiography was useful in the diagnosis of CO2 embolism and to assess response to inhaled epoprostenol. PMID- 12651644 TI - Parental intervention choices for children undergoing repeated surgeries. AB - No studies have examined parental preference for a preoperative intervention in healthy children undergoing subsequent surgeries. We collected data prospectively from 83 children who previously underwent surgery and were part of an investigation by our study group, then returned for a subsequent surgery. At the initial surgery, children were assigned (no parental intervention) to receive oral midazolam (n = 13), or parental presence during the induction of anesthesia (PPIA, n = 27), or PPIA + midazolam (n = 10) or no intervention (n = 33). At a subsequent surgery, parents chose the preoperative intervention. We found that >80% of all parents chose PPIA (with or without midazolam) at the subsequent surgery regardless of the intervention they received previously. Of parents whose children received PPIA at the initial surgery, 70% chose PPIA again. In contrast, only 23% of the patients who received midazolam at the initial surgery requested midazolam at the subsequent surgery and only 15% of the patients who received no intervention at the initial surgery requested no intervention at the subsequent surgery. All parents of very anxious children at the initial surgery chose some intervention at the subsequent surgery (P = 0.022). Parents of children who underwent a subsequent surgery preferred PPIA regardless of any previous intervention. Also, parents' intervention preferences at the subsequent surgery were influenced by children's anxiety at the initial surgery. IMPLICATIONS: Parents of children who undergo a subsequent surgery prefer to be present during the induction of anesthesia regardless of whether the child was medicated or had parents present or did not receive anything at the initial surgery. Also, parents' preference for medication or parental presence at the subsequent surgery was influenced by the child's anxiety at the initial surgery. PMID- 12651645 TI - Clonidine facilitates controlled hypotension in adolescent children. AB - In this randomized, double-blinded, placebo-controlled trial, we investigated whether clonidine lessened the requirements for isoflurane, fentanyl, and labetalol to provide controlled hypotension in children who underwent oromaxillofacial surgery. We also studied preoperative sedative effects, the hemodynamic response to nasotracheal intubation, and recovery characteristics. Thirty-nine healthy children, aged 10-16 yr, received clonidine 5 micro g/kg or placebo on the night before surgery and 90 min before surgery. By self assessment, children in both groups showed similar anxiety, whereas observers rated clonidine patients as more sedated (P < 0.01). Heart rate during induction remained significantly decreased in clonidine patients compared with placebo patients (P < 0.001), as did arterial blood pressure before induction (P < 0.01) and peak pressure after intubation (P < 0.001). Children who took clonidine required significantly less isoflurane to maintain a mean arterial blood pressure of 60 +/- 4 mm Hg (mean isoflurane concentration, clonidine 0.99% versus placebo 1.33; P = 0.0004) and required less fentanyl than placebo patients (P = 0.002). Fewer treatment patients received labetalol (clonidine, n = 3 versus placebo, n = 13; P = 0.004). There was a trend toward faster recovery in the clonidine group, with a shortened recovery room stay (P = 0.03). We conclude that clonidine is a useful adjunct for controlled hypotension in children. IMPLICATIONS: This study shows that adolescents having major jaw surgery are helped by the blood pressure decreasing drug clonidine. This drug allows smaller doses of anesthetics, pain relievers, and blood pressure-decreasing drugs to be used; reduces changes in heart rate and blood pressure; and provides faster recovery from the anesthetic. PMID- 12651646 TI - Suprascapular nerve block prolongs analgesia after nonarthroscopic shoulder surgery but does not improve outcome. AB - Suprascapular nerve block (SSNB) reportedly improves analgesia and 24-h outcomes after arthroscopic shoulder surgery performed under general anesthesia. In this study, we assessed the analgesic and clinical outcome efficacy of SSNB as an adjunct to interscalene brachial plexus block (ISB) for ambulatory nonarthroscopic shoulder surgery. Fifty patients were randomized to receive either a SSNB or sham injection as part of a standardized ISB-general anesthesia regimen. Time to first significant pain (the primary outcome measure) was significantly delayed in the SSNB group (594 +/- 369 min versus 375 +/- 273 min, respectively; P = 0.02). There were no other differences between groups with regard to postanesthesia recovery unit measures, 24-h assessment of pain, supplemental analgesic use, or quality of life outcomes. We conclude that adjunctive SSNB adds minimal value to a primary ISB anesthetic for nonarthroscopic shoulder surgery. IMPLICATIONS: When used as an adjunct to an interscalene block combined with general anesthesia, suprascapular nerve block with bupivacaine moderately prolongs analgesia without improving other outcome measures after ambulatory nonarthroscopic shoulder surgery. PMID- 12651647 TI - Costs and effectiveness of rofecoxib, celecoxib, and acetaminophen for preventing pain after ambulatory otolaryngologic surgery. AB - We designed this randomized, double-blinded, placebo-controlled study to compare the analgesic effect of the cyclooxygenase-2 inhibitors rofecoxib and celecoxib with acetaminophen when administered before outpatient otolaryngologic surgery in 240 healthy subjects. Patients were assigned to one of four study groups: Group 1, control (vitamin C 500 mg); Group 2, acetaminophen 2 g; Group 3, celecoxib 200 mg; or Group 4, rofecoxib 50 mg. The first oral dose of the study medication was administered 15-45 min before surgery, and a second dose of the same medication was given on the morning after surgery. Recovery times, side effects, pain scores, and the use of rescue analgesics were recorded. Follow-up evaluations were performed at 24 and 48 h after surgery to assess postdischarge pain, analgesic requirements, nausea, and patient satisfaction with their postoperative pain management and quality of recovery. The need for rescue analgesia and peak pain scores were used as the primary end points for estimating efficacy, and the costs to achieve complete satisfaction with analgesia were used for the cost efficacy comparisons. Premedication with oral rofecoxib (50 mg) or celecoxib (200 mg) was more effective than placebo in reducing postoperative pain scores and analgesic requirements in the postoperative care unit and after discharge. The analgesic efficacy of oral acetaminophen (2 g) was limited to the postdischarge period. Patient satisfaction with pain management was improved in all three treatment groups compared with placebo but was higher with celecoxib and rofecoxib compared with acetaminophen. Rofecoxib was also more effective than celecoxib in reducing pain and improving patient satisfaction after otolaryngologic surgery. Rofecoxib achieved complete satisfaction with pain control in one additional patient, who would not have otherwise been satisfied, at lower incremental costs to the institution compared with celecoxib. We conclude that rofecoxib 50 mg orally is more cost-effective for reducing postoperative pain and improving patient satisfaction with their postoperative pain management than celecoxib (200 mg) or acetaminophen (2 g) in the ambulatory setting. IMPLICATIONS: Oral premedication with rofecoxib (50 mg) was more effective than celecoxib (200 mg) and acetaminophen (2 g) in reducing postoperative pain and in improving the quality of recovery and patient satisfaction with pain management after outpatient otolaryngologic surgery with only a small increase in cost of care. PMID- 12651648 TI - Ginger does not prevent postoperative nausea and vomiting after laparoscopic surgery. AB - IMPLICATIONS: The potential antiemetic effect of two different oral doses of the herbal remedy ginger (Zingiber officinale) to prevent postoperative nausea and vomiting in 180 patients undergoing gynecologic laparoscopy was investigated in this randomized, double-blinded trial. Ginger failed to reduce the incidence of postoperative nausea and vomiting after these procedures. PMID- 12651649 TI - Isoflurane action in spinal cord indirectly depresses cortical activity associated with electrical stimulation of the reticular formation. AB - Anesthetics act in the spinal cord to ablate both movement and the ascending transmission of nociceptive information. We investigated whether a spinal cord action of isoflurane affected cortical activity as determined by the electroencephalogram desynchronization that occurs after electrical stimulation of the midbrain reticular formation (MRF). Six goats were anesthetized with isoflurane, and neck dissections were performed to permit differential isoflurane delivery to the head and torso. The electroencephalogram was recorded before, during, and after focal electrical stimulation (0.05, 0.1, 0.2, 0.3, and 0.4 mA) in the MRF; in each animal, the brain isoflurane was maintained constant ( approximately 1%). When the torso isoflurane was 0.3% +/- 0.1%, the spectral edge frequency after MRF electrical stimulation (15.3 +/- 1.7 Hz, averaged across all stimulus currents) was more than the spectral edge frequency when the torso isoflurane was 1.2% +/- 0.2% (12.9 +/- 1.0 Hz, averaged across all stimulus currents; P < 0.05). Bispectral index values were similarly affected: 60 +/- 6 when torso isoflurane was low versus 53 +/- 7 at high torso isoflurane (P < 0.05). These results suggest that a spinal depressant action of isoflurane on ascending somatosensory transmission can modulate reticulo-thalamocortical arousal mechanisms, hence possibly reducing anesthetic requirements for unconsciousness and amnesia. IMPLICATIONS: Isoflurane action in the spinal cord indirectly reduces the cortical activity associated with electrical stimulation of the reticular formation, an effect that might contribute to anesthetic-induced amnesia and unconsciousness. PMID- 12651650 TI - The memory effects of general anesthesia persist for weeks in young and aged rats. AB - Studies demonstrate lasting cognitive impairment in elderly persons after anesthesia and surgery. We tested the hypothesis that general anesthesia contributes to this cognitive impairment. Six- and 18-mo-old Fischer 344 rats were trained in a 12-arm radial arm maze and were then randomized to anesthesia for 2 h with 1.2% isoflurane/70% nitrous oxide/30% oxygen or a control treatment consisting of 30% oxygen. Rats recovered for 24 h and then were tested daily on the radial arm maze for 8 wk. Performance of young control rats was stable throughout the experiment. In contrast, aged control rats improved their performance as measured by time to complete the maze but not by error rate. After anesthesia, time to complete the maze did not change in young rats, but error rate decreased (P < 0.05 at 1 and 3 wk), indicating improved performance. In contrast, previously anesthetized aged rats failed to improve with repeated testing and took longer to complete the maze than aged control rats (P < 0.05 at 1 and 3 wk). These data demonstrate that general anesthesia with isoflurane and nitrous oxide improves the memory performance on an established spatial memory task in young rats, but in aged rats it attenuates the improvement in performance that otherwise occurs with repeated testing. Therefore, isoflurane and nitrous oxide anesthesia produces a sustained learning impairment in aged rats. IMPLICATIONS: This study demonstrates that general anesthesia with isoflurane and nitrous oxide improves spatial memory in young rats but impairs it in aged rats for at least 3 wk, indicating that it can influence memory for much longer than previously recognized and may adversely affect memory processes in the aged. PMID- 12651651 TI - Isoflurane antagonizes the capacity of flurothyl or 1,2 dichlorohexafluorocyclobutane to impair fear conditioning to context and tone. AB - In animals, the conventional inhaled anesthetic, isoflurane, impairs learning fear to context and fear to tone, doing so at concentrations that produce amnesia in humans. Nonimmobilizers are inhaled compounds that do not produce immobility in response to noxious stimulation, nor do they decrease the requirement for conventional inhaled anesthetics. Like isoflurane, the nonimmobilizer 1,2 dichlorohexafluorocyclobutane (2N) impairs learning at concentrations less than those predicted from its lipophilicity to produce anesthesia. The capacity of the nonimmobilizer di-(2,2,2,-trifluoroethyl) ether (flurothyl) to affect learning and memory has not been studied. Both nonimmobilizers can cause convulsions. We hypothesized that if isoflurane, 2N, and flurothyl act by the same mechanism to impair learning and memory, their effects should be additive. We found that isoflurane, 2N, and flurothyl (each, alone) impaired learning fear to context and fear to tone in rats, with the nonimmobilizers doing so at concentrations less than those that cause convulsions. (Fear was defined by freezing [volitional immobility] in the presence of the conditioned stimulus [context or tone].) However, the combination of isoflurane and 2N or flurothyl produced an antagonistic rather than an additive effect on learning, a finding in conflict with our hypothesis. And flurothyl was no less potent than 2N (at least no less potent relative to the concentration of each that produced convulsions) in its capacity to impair learning. We conclude that conventional inhaled anesthetics and nonimmobilizers impair learning and memory by different mechanisms. The basis for this impairment remains unknown. IMPLICATIONS: Conventional inhaled anesthetics and nonimmobilizers are antagonistic in their effects on learning and memory, and this finding suggests that they impair learning and memory, at least in part, by different mechanisms. PMID- 12651652 TI - Blockade of voltage-operated neuronal and skeletal muscle sodium channels by S(+) and R(-)-ketamine. AB - Besides its general anesthetic effect, ketamine has local anesthetic-like actions. We studied the voltage- and use-dependent interaction of S(+)- and R(-) ketamine with two different isoforms of voltage-operated sodium channels, with a special emphasis on the difference in affinity between resting and inactivated channel states. Rat brain IIa and human skeletal muscle sodium channels were heterologously expressed in human embryonic kidney 293 cells. S(+)- and R(-) ketamine reversibly suppressed whole-cell sodium inward currents; the 50% inhibitory concentration values at -70 mV holding potential were 240 +/- 60 microM and 333 +/- 93 microM for the neuronal isoform and 59 +/- 10 microM and 181 +/- 49 microM for the skeletal muscle isoform. S(+)-ketamine was significantly more potent than R(-)-ketamine in the skeletal muscle isoform only. Ketamine had a higher affinity to inactivated than to resting channels. However, the estimated difference in affinity between inactivated and resting channels was only 8- to 10-fold, and the time course of drug equilibration between inactivated and resting channels was too fast to cause use-dependent block at 10 Hz up to a concentration of 300 microM. These results suggest that ketamine is less effective than lidocaine-like local anesthetics in stabilizing the inactivated channel state. IMPLICATIONS: Blockade of sodium channels by ketamine shows voltage dependency, an important feature of local anesthetic action. However, ketamine is less effective than lidocaine-like local anesthetics in stabilizing the inactivated state. Because it does not elicit phasic blockade at small concentrations, its ability to reduce the firing frequency of action potentials may be small. PMID- 12651654 TI - The effect of inhaled colforsin daropate on contractility of fatigued diaphragm in dogs. AB - We studied the effect of inhaled colforsin daropate, a water-soluble forskolin derivative, on the contractility of fatigued diaphragm in dogs. Animals were divided into 3 groups of 8. In each group, diaphragmatic fatigue was induced by intermittent supramaximal bilateral electrophrenic stimulation at a frequency of 20-Hz stimulation applied for 30 min. Immediately after the end of the fatigue producing period, Group 1 received inhaled vehicle, Group 2 received inhaled colforsin daropate 0.1 mg/mL, and Group 3 received inhaled colforsin daropate 0.2 mg/mL. We assessed diaphragmatic contractility by transdiaphragmatic pressure (Pdi). After fatigue was produced, in each group, Pdi at low-frequency (20-Hz) stimulation decreased from baseline values (P < 0.05), and there was no change in Pdi at high-frequency (100-Hz) stimulation. In Groups 2 and 3, during colforsin daropate inhalation, Pdi at both stimuli increased from fatigued values (P < 0.05). The increase in Pdi was significantly larger in Group 3 than in Group 2. The integrated electrical activity of the diaphragm did not change in any group. We conclude that inhaled colforsin daropate causes an increase in contractility of fatigued canine diaphragm in a dose-related fashion. IMPLICATIONS: Diaphragmatic fatigue may contribute to the development of respiratory failure. Inhaled colforsin daropate improves, in a dose-dependent manner, the contractility of fatigued diaphragm in dogs. PMID- 12651653 TI - 5-HT3 receptors partially mediate halothane depression of spinal dorsal horn sensory neurons. AB - We recently reported that gamma-aminobutyric acid type A- and strychnine sensitive glycine receptor systems partially mediate halothane depression of spinal dorsal horn low-threshold neurons. Serotonin subtype 3 (5-HT(3)) receptors belong to the same ligand-activated ion-channel family as gamma-aminobutyric acid type A- and strychnine-sensitive glycine receptors, so we examined the possible involvement of 5-HT receptor systems in halothane depression of spinal sensory neurons. Extracellular recordings of spinal low-threshold neurons were obtained in decerebrate, spinally transected rats. Receptive field size and brush-induced activity were recorded in the presence or absence of 5-HT antagonists and in the presence or absence of 1.1% (1 minimum alveolar anesthetic concentration) halothane. In the absence of halothane, antagonists had no effect on receptive field size or brush-induced activity. In the presence of halothane, methysergide, a nonselective 5-HT antagonist, and tropisetron, a selective 5-HT(3) antagonist, significantly reversed the halothane-induced reduction in receptive field size but did not alter halothane depression of brush-induced activity. Methiothepin, a 5-HT(1) antagonist, and ketanserin, a 5-HT(2) antagonist, did not reverse halothane depression. These results support the hypothesis that 5-HT(3) receptors partially mediate some inhibitory effects of halothane on spinal dorsal horn neurons. IMPLICATIONS: The results of this study support the hypothesis that halothane depression of spinal sensory neuronal responses to low-intensity stimuli is mediated, to a minor extent, by serotonin subtype 3 neurotransmitter systems. PMID- 12651655 TI - The effects of propofol and etomidate on airway contractility in chronically hypoxic rats. AB - We investigated the effect of two IV anesthetics, propofol and etomidate, on airway responsiveness in a rat model of chronic hypoxia (CH) in comparison with normoxic rats. CH rats were obtained using a hypobaric chamber (14 days at a barometric pressure of 380 mm Hg). The ability of both anesthetics to relax and prevent agonist-induced contraction was assessed in isolated tracheal rings precontracted with the muscarinic agonist carbachol (CCh) and the depolarizing agent KCl. Cumulative concentrations of both compounds relaxed tracheal rings precontracted with CCh or KCl with a similar amplitude in CH and normoxic rats. In tracheal rings precontracted with CCh, the negative logarithm of anesthetics that reduced the maximal contraction by 30%, i.e., -log half-maximal inhibitory concentration, for propofol and etomidate were 4.10 +/- 0.09 and 4.12 +/- 0.15 in normoxic rats and 4.20 +/- 0.22 and 3.61 +/- 0.19 in CH rats, respectively. At a fixed concentration, propofol (3 x 10(-4) M) or etomidate (10(-4) M) also inhibited CH tracheal rings contraction in response to cumulative concentrations of CCh and KCl. However, in contrast with the equivalent relaxant effect of both anesthetics, etomidate was two-fold less effective than propofol for inhibiting the subsequent contraction to CCh and KCl. These results indicate that propofol and etomidate retain their relaxant properties in CH rat airways by acting on the pharmaco- and electromechanical coupling. IMPLICATIONS: Anesthesia may cause airway constriction or bronchospasm in patients with normal or pathological airways. This study investigated the ability of propofol and etomidate to both reverse precontraction and inhibit contraction of tracheal rings isolated from chronically hypoxic rats. PMID- 12651656 TI - The dose effect of ephedrine on the onset time of vecuronium. AB - A small dose of ephedrine decreases the onset time of rocuronium and cisatracurium; however, ephedrine might be associated with adverse hemodynamic effects. The appropriate dose of ephedrine has not been determined. We, therefore, studied 120 patients anesthetized with fentanyl 2 microg/kg and propofol 2-2.5 mg/kg who were randomly divided to receive either ephedrine (30, 70, or 110 microg/kg) or saline. During propofol anesthesia, the neuromuscular block was monitored by mechanomyography by using submaximal current of train-of four stimulation every 10 s. To determine cardiac output, a transcutaneous Doppler probe was placed externally at the suprasternal notch. Tracheal intubation was performed by a blinded investigator at 2 min after vecuronium. Neuromuscular block, intubating conditions, and hemodynamic effects were measured during the induction of anesthesia. Both ephedrine 70 and 110 microg/kg improved intubating conditions at 2 min after vecuronium; however, 110 microg/kg was associated with adverse hemodynamic effects. We conclude that ephedrine 70 microg/kg given before the induction of anesthesia improved intubating conditions at 2 min after vecuronium, probably by increased cardiac output without significant adverse hemodynamic effects. IMPLICATIONS: Ephedrine 70 microg/kg given before the induction of anesthesia improved tracheal intubating conditions at 2 min after vecuronium by increased cardiac output without significant adverse hemodynamic effects. PMID- 12651657 TI - The effect of remifentanil on seizure duration and acute hemodynamic responses to electroconvulsive therapy. AB - We designed this prospective, randomized, double-blinded, placebo-controlled, crossover study to evaluate the effect of different doses of remifentanil on the acute hemodynamic response and duration of seizure activity after a standardized electroconvulsive therapy (ECT) stimulus. Twenty consenting patients with major depressive disorders receiving maintenance ECT participated in this study. Eighty ECT treatments were evaluated. All patients were premedicated with glycopyrrolate 0.2 mg IV, unconsciousness was induced with methohexital 1 mg/kg IV, and muscle paralysis was produced with succinylcholine 1.2 mg/kg IV. Subsequently, patients received 1 of 3 different doses of remifentanil 25, 50, and 100 microg or saline (control) in a random sequence immediately after methohexital at 4 consecutive ECT treatments. Labetalol, in 5-mg IV boluses, was used as a rescue antihypertensive medication. A fixed suprathreshold electrical stimulus was administered to elicit a seizure, and the times from the stimulus to the cessation of the motor and electroencephalographic (EEG) seizure activity were noted. Pre- and post-ECT blood pressure values were significantly decreased in the 100- microg remifentanil group compared with the control group. The durations of motor (38 +/- 9 s to 43 +/- 15 s) and EEG (55 +/- 29 s to 60 +/- 21 s) seizure activity were not significantly different among the four groups. Similarly, recovery times to eye opening, obeying commands, and discharge from the recovery room did not differ among the four study groups. The requirement for labetalol after ECT was nonsignificantly decreased in the remifentanil groups. In conclusion, remifentanil 100 microg IV attenuated the acute hemodynamic response to ECT. Furthermore, remifentanil had no adverse effect on the duration of ECT induced seizure activity. Finally, adjunctive use of remifentanil did not prolong recovery times or increase post-ECT side effects. IMPLICATIONS: Remifentanil (100 microg IV) attenuated the acute hemodynamic response after electroconvulsive therapy (ECT) without adversely affecting the length of the ECT-induced seizure activity or prolonging recovery times. PMID- 12651658 TI - Anaphylactic reactions to isosulfan blue dye during sentinel node lymphadenectomy for breast cancer. AB - IMPLICATIONS: Intraoperative use of isosulfan dye for lymphatic mapping may result in anaphylaxis. Furthermore, in some patients, intravascular absorption of isosulfan may induce serum discoloration causing interference with pulse oximetry function. PMID- 12651659 TI - Prolonged dexmedetomidine infusion as an adjunct in treating sedation-induced withdrawal. AB - IMPLICATIONS: Dexmedetomidine, an alpha(2)-adrenoceptor agonist, is indicated for sedating patients on mechanical ventilation. It has been approved by the Food and Drug Administration for 24-h use. This is a report concerning a patient in whom a continuous infusion of dexmedetomidine was safely used for a week to help in averting frank withdrawal symptoms from an opioid and benzodiazepines. PMID- 12651660 TI - The anesthetic conserving device compared with conventional circle system used under different flow conditions for inhaled anesthesia. AB - The Anesthetic Conserving Device (ACD) is a high-flow anesthesia system closed to volatile anesthetics only. We compared the ACD with a circle system under different fresh gas flow (FGF) conditions. Eighty-one patients undergoing major surgery were randomly allocated to receive sevoflurane from a circle circuit combined either with the ACD placed at the Y-piece (n = 41) or with a vaporizer (n = 40). The FGF was set to 8 L/min in the ACD system, where the circle circuit served as a nonrebreather. In the conventional circle system without ACD, the vaporizer was supplied with 1-, 1.5-, 3-, and 6-L/min FGFs. We compared the ACD with the circle system under the four FGFs in terms of sevoflurane dosing, sevoflurane consumption, humidification efficiency, and environmental pollution. The ACD and the low-flow circle system (1.5- and 1-L/min FGFs) resulted in the smallest sevoflurane consumption. The increase in inspired sevoflurane concentration was faster with the circle system than with the ACD only with FGFs > or =3 L/min. The removal of ACD from the circuit allowed the fastest washout of sevoflurane. Respiratory gas humidification was always adequate. Sevoflurane ambient concentration with the ACD was 1-70 ppb. The ACD is a valid and simple alternative to low-flow systems. IMPLICATIONS: The Anesthetic Conserving Device (ACD) is a new device for anesthetic vapor delivery. We demonstrated that the ACD reduces anesthetic consumption and environmental pollution similarly to a low flow circle system, offering advantages such as simplicity, no toxicity from compounds produced in the absorber, and potential cost savings. PMID- 12651661 TI - Bispectral index-derived facial electromyography-guided fentanyl titration in the opiate-exposed patient. AB - IMPLICATIONS: A 34-yr-old man, possibly opiate tolerant, underwent the second part of a scoliosis repair. We describe a narcotic titration protocol by using facial electromyography data derived from the bispectral index monitor that resulted in a good clinical outcome. PMID- 12651662 TI - Nifedipine potentiates the antinociceptive effect of endomorphin-1 microinjected into the periaqueductal gray in rats. AB - Endomorphin-1 is a novel endogenous mu-opioid ligand. We investigated the antinociceptive interaction between endomorphin-1 and nifedipine, an L-type calcium channel blocker, microinjected into the midbrain ventrolateral periaqueductal gray (vPAG), using the spinally-organized tail-flick test and the supraspinally-organized tail-pressure test in rats. Sprague-Dawley rats were stereotaxically implanted with a guide cannula lowered into the vPAG. Microinjection of endomorphin-1 into the vPAG led to dose-related increases in antinociceptive responses in the tail-flick test and tail-pressure test. Pretreatment with the mu-opioid receptor-selective antagonist beta-funaltrexamine blocked the antinociceptive effect of endomorphin-1. Pretreatment with beta funaltrexamine alone had no effect on the tail-flick latency and tail-pressure threshold. Microinjection of nifedipine alone into the vPAG did not produce an antinociceptive response in the tail-flick test and tail-pressure test. However, injection of nifedipine into the vPAG potentiated the antinociceptive effect of endomorphin-1, producing a significant leftward shift in the dose-response curve of endomorphin-1 in both the tail-flick and tail-pressure tests. This result shows that the potent antinociceptive effect of endomorphin-1 microinjected into the vPAG is mediated through the mu-opioid receptor and is potentiated by concomitant administration of nifedipine. IMPLICATIONS: This study shows that the potent antinociceptive effect of endomorphin-1 microinjected into the ventrolateral periaqueductal gray is potentiated by concomitant administration of nifedipine. This suggests that calcium channel blockers may enhance the analgesia of opioids in patients with calcium channel blocker treatment. PMID- 12651663 TI - Antinociception with intrathecal alpha-methyl-5-hydroxytryptamine, a 5 hydroxytryptamine 2A/2C receptor agonist, in two rat models of sustained pain. AB - Type 2 serotonin (5-hydroxytryptamine [5-HT](2)) receptors in the spinal cord have been reported to mediate antinociception using pain threshold tests, but little is known about the actions of spinal 5-HT(2) receptors in sustained pain. In rats, we examined antinociceptive effects of the intrathecal administration of a 5-HT(2A/2C) receptor agonist, alpha-methyl-5-HT maleate (alpha-m-5-HT), using the formalin test and the chronic constriction injury (CCI) model. An intrathecal catheter was implanted for injection of drugs. In the formalin test, flinches were counted from Minute 1 to 2 and Minute 5 to 6 (Phase 1) and then for 1-min periods at 5-min intervals from 10 to 60 min (Phase 2). In rats with CCI, hind paw withdrawal latency after thermal stimulation was measured. In the formalin test, intrathecal administration of alpha-m-5-HT (1 to 100 microg) dose dependently suppressed the number of flinches in both Phases 1 and 2. In the CCI model, intrathecally administered alpha-m-5-HT (10 to 100 microg) attenuated thermal hyperalgesia in a dose-dependent manner. These effects were reversed by intrathecal pretreatment with a 5-HT(2A/2C) antagonist, ketanserin (30 microg), or a muscarinic receptor antagonist, atropine (30 microg). These findings suggest that spinal 5-HT(2A/2C) receptors mediate antinociception in inflammatory pain and neuropathic pain, and the muscarinic receptors contribute to this action. IMPLICATIONS: Activation of spinal 5-hydroxytryptamine(2A/2C) receptors mediate antinociception in rat-sustained pain models such as inflammatory pain and neuropathic pain, and spinal muscarinic receptors are involved in this action. PMID- 12651664 TI - A small preoperative test dose of intravenous fentanyl can predict subsequent analgesic efficacy and incidence of side effects in patients due to receive epidural fentanyl. AB - Because individual variation is a likely factor affecting both the incidence and severity of side effects and the analgesic efficacy of epidural opioids, assessment of individual variation could be useful in deciding optimal dosage. By evaluating the response to a small test dose of IV fentanyl, we designed this study to predict the degree of pain relief and the incidence of side effects in patients who would be receiving postoperative epidural fentanyl. Before the induction of anesthesia, 50 micro g of fentanyl was administered IV, and 2 min after fentanyl, the patient response was evaluated. Twenty-three patients, who reported nausea, sleepiness, dizziness, sensation of warmth, and other symptoms, were categorized as responders (Group R); the remaining 20 patients were categorized as nonresponders (Group NR). At the completion of surgery, infusion of epidural fentanyl was administered (0.3 mg/d in 0.25% bupivacaine) for 96 h. At postoperative Hours 6 and 24, Group R had significantly lower visual analog scale scores for postoperative pain intensity and required fewer analgesics than Group NR. The incidence of side effects, however, was 74% for Group R and 10% for Group NR (P < 0.05), and side effects were more serious in Group R. This study demonstrates that preoperative administration of a small dose of fentanyl during the induction of anesthesia enables prediction of the analgesic efficacy of postoperative epidural fentanyl and the incidence and severity of side effects. IMPLICATIONS: Preoperative administration of a small dose of fentanyl during the induction of anesthesia enables prediction of the analgesic efficacy of postoperative epidural fentanyl and the incidence and severity of side effects. PMID- 12651665 TI - Intrathecal clonidine added to a bupivacaine-morphine spinal anesthetic improves postoperative analgesia for total knee arthroplasty. AB - Postoperative pain after total knee arthroplasty (TKA) is severe and can complicate early physical therapy. We tested the hypothesis that intrathecal clonidine would improve postoperative analgesia for TKA using a hyperbaric bupivacaine spinal anesthetic. In a double-blinded, placebo-controlled protocol, 81 ASA physical status I-III patients undergoing either a single or bilateral TKA were randomized into 4 groups with the following 2-mL solutions added to 15 mg of hyperbaric bupivacaine: 1) sterile saline, 2) morphine (250 microg), 3) morphine (250 microg) with clonidine (25 microg), and 4) morphine (250 microg) with clonidine (75 microg). At 1, 2, 4, 6, 12, and 24 h postoperatively, we measured visual analog scales (VAS), cumulative IV morphine consumption, hemodynamics, nausea, ancillary drugs, and side effects. Our primary comparison was between the clonidine with morphine groups versus the morphine group. We found that the combined administration of intrathecal clonidine and morphine decreased 24 h IV morphine consumption by 13 mg (P = 0.028) when compared with intrathecal morphine alone. This corresponded to a decrease in the VAS score of 1.3 cm at 24 h postoperatively (P = 0.047). Adverse side effects were similar among all groups with the exception of more relative hypotension in the clonidine groups through postoperative hour 6. We conclude that the coadministration of intrathecal clonidine and morphine decreases the 24-h IV morphine consumption and improves the 24-h VAS score when compared with intrathecal morphine alone. IMPLICATIONS: In this prospective, randomized, double-blinded, and placebo-controlled trial, we identify an effective postoperative analgesic approach in total knee replacement surgery. Intrathecal morphine (250 microg) combined with clonidine (25 or 75 microg) provided superior analgesia compared with intrathecal morphine alone. PMID- 12651666 TI - Continuous interscalene brachial plexus block for postoperative pain control at home: a randomized, double-blinded, placebo-controlled study. AB - In this study, we investigated the efficacy of patient-controlled regional analgesia for outpatients undergoing moderately painful orthopedic surgery of the shoulder. Preoperatively, patients (n = 20) received an interscalene nerve block and perineural catheter. Postoperatively, patients were discharged home with both oral opioids and a portable infusion pump delivering either 0.2% ropivacaine or 0.9% saline, determined randomly in a double-blinded manner. Daily end points included pain scores, opioid use and side effects, sleep quality, and technique complications. Ropivacaine (n = 10) infusion significantly reduced pain compared with saline (n = 10) infusion. The average pain at rest (scale: 0-10) on postoperative day 1 (median, 25th-75th percentiles) was 4.8 (4.0-5.0) for the saline group, versus 0.0 (0.0-2.0) for the ropivacaine group (P < 0.001). Oral opioid use and related side effects were also significantly decreased in the ropivacaine group. On postoperative day 1, median tablet consumption was 8.0 (6.5 9.5) and 0.5 (0.0-1.0) for the saline and ropivacaine groups, respectively (P < 0.001). Sleep disturbance scores were nearly threefold greater on the first postoperative night for patients receiving saline (P = 0.013). We conclude that after moderately painful orthopedic surgery of the shoulder, ropivacaine infusion using a portable infusion pump and an interscalene perineural catheter at home decreased pain, opioid use and related side effects, and sleep disturbances. IMPLICATIONS: This randomized, double-blinded, placebo-controlled study demonstrated that ropivacaine, infused with a portable infusion pump via an interscalene perineural catheter for 3 days at home, significantly decreased postoperative pain after orthopedic surgery of the shoulder. In addition to providing potent analgesia and increasing patient satisfaction, perineural infusion decreased opioid requirements and their associated side effects. PMID- 12651667 TI - The role of interleukin-6 in nociception and pain. AB - IMPLICATIONS: That IL-6 is an interesting target in the study of pain is underscored by its biomolecular properties, its localization after experimental pain, and its modulating effect on pain after administration. PMID- 12651668 TI - Sustained increases in productivity with maintenance of quality in an academic anesthesia practice. AB - Previously, the authors reported trends in anesthesia quality and productivity in a university-based anesthesia practice as it responded to increasing service demands with shortages of qualified staff and decreasing reimbursement. From 1992 to 1997, productivity increased, with a significant decrease in patient injury. In this study, we analyzed whether previous productivity and quality gains were sustained from 1998 to 2000 despite continued staff shortages. Productivity, caseload, and outcome data were abstracted from departmental administrative and quality-improvement reports. Retrospective cohort analysis compared trends during 3 yr of moderate productivity (1994-1996) with those during 3 yr of high productivity (1998-2000). The mean monthly productivity in 1998-2000 (15 +/- 0.6 billed hours per attending per clinical day) was larger than levels from 1994 to 1996 (mean, 14 +/- 0.7 h; P < 0.01). The overall continuous quality improvement report rate was slower at larger productivity levels, as were rates of patient injuries. When adjusted for declining report rates, patient injury rates showed no change between smaller- and larger-productivity years. Adjusted rates of operational inefficiencies and human errors were more frequent at larger productivity levels. Although the pressures of increased demands, shrinking resources, and shortages of qualified academic anesthesiologists have not abated, productivity and quality have been sustained. Future management must be directed toward reductions in operational inefficiencies and human error. IMPLICATIONS: Our academic anesthesia service sustained increases in productivity with maintenance of quality. During a 3-yr period of high productivity, patient injury rates did not increase compared with prior years with lower productivity. PMID- 12651669 TI - Labor costs incurred by anesthesiology groups because of operating rooms not being allocated and cases not being scheduled to maximize operating room efficiency. AB - Determination of operating room (OR) block allocation and case scheduling is often not based on maximizing OR efficiency, but rather on tradition and surgeon convenience. As a result, anesthesiology groups often incur additional labor costs. When negotiating financial support, heads of anesthesiology departments are often challenged to justify the subsidy necessary to offset these additional labor costs. In this study, we describe a method for calculating a statistically sound estimate of the excess labor costs incurred by an anesthesiology group because of inefficient OR allocation and case scheduling. OR information system and anesthesia staffing data for 1 yr were obtained from two university hospitals. Optimal OR allocation for each surgical service was determined by maximizing the efficiency of use of the OR staff. Hourly costs were converted to dollar amounts by using the nationwide median compensation for academic and private-practice anesthesia providers. Differences between actual costs and the optimal OR allocation were determined. For Hospital A, estimated annual excess labor costs were $1.6 million (95% confidence interval, $1.5-$1.7 million) and $2.0 million ($1.89-$2.05 million) when academic and private-practice compensation, respectively, was calculated. For Hospital B, excess labor costs were $1.0 million ($1.08-$1.17 million) and $1.4 million ($1.32-1.43 million) for academic and private-practice compensation, respectively. This study demonstrates a methodology for an anesthesiology group to estimate its excess labor costs. The group can then use these estimates when negotiating for subsidies with its hospital, medical school, or multispecialty medical group. IMPLICATIONS: We describe a new application for a previously reported statistical method to calculate operating room (OR) allocations to maximize OR efficiency. When optimal OR allocations and case scheduling are not implemented, the resulting increase in labor costs can be used in negotiations as a statistically sound estimate for the increased labor cost to the anesthesiology department. PMID- 12651670 TI - Determining optimum operating room utilization. AB - Economic considerations suggest that it is desirable to keep operating rooms fully used when staffed, but the optimum utilization of an operating room (OR) is not known. We created a simulation of an OR to define optimum utilization. We set operational goals of having cases start within 15 min of the scheduled time and of having the cases end no more than 15 min past the scheduled end of the day. Within these goals, a utilization of 85% to 90% is the highest that can be achieved without delay or running late. Increasing the variability of case duration decreases the utilization that can be achieved within these targets. IMPLICATIONS: Using a simulated operating room (OR), the authors demonstrate that OR utilization higher than 85% to 90% leads to patient delays and staff overtime. Increased efficiency of an OR comes at a cost of patient convenience. PMID- 12651671 TI - Cerebral blood flow is not altered in sheep with Pseudomonas aeruginosa sepsis treated with norepinephrine or nitric oxide synthase inhibition. AB - The origin of cerebral dysfunction in patients with sepsis is still unclear. However, altered cerebral perfusion may play an important role in its pathogenesis. Using an established, chronic model of hyperdynamic ovine sepsis, we examined cerebral perfusion in 20 sheep subjected to a continuous infusion of live Pseudomonas aeruginosa. After 24 h of sepsis, the hypotensive sheep (reduction in mean arterial blood pressure by 16%; P < 0.05) received the nitric oxide synthase inhibitor N(G)-mono-methyl-L-arginine (L-NMMA; 7 mg. kg(-1). h( 1); n = 7), norepinephrine (NE; n = 7), or normal saline (control; n = 6). NE infusion was individually targeted to achieve the same increase in mean arterial blood pressure as that observed in matched sheep of the L-NMMA group. Regional perfusion was measured by using colored microspheres. Although L-NMMA caused a significant increase in systemic vascular resistance index (1167 +/- 104 versus 793 +/- 59 dyne. cm(-5). m(2); P < 0.05), it caused a change neither in cerebrovascular resistance nor in cerebral blood flow. When related to systemic blood flow, a redistribution of blood flow to the brain became obvious. The NE associated increase in systemic blood pressure (98 +/- 5 versus 83 +/- 5; P < 0.05) was accompanied by an increase in cardiac output (7.8 +/- 0.5 versus 6.7 +/ 0.6; P < 0.05) and, hence, systemic perfusion. However, blood flow to the brain remained unaffected. Although detrimental vasoconstrictive effects of NE and L NMMA, including cerebral hypoperfusion, are discussed, neither drug had any effect on cerebral perfusion during experimental hyperdynamic sepsis. IMPLICATIONS: Cerebral dysfunction is often found in septic patients. In this regard, it is debated whether vasopressor drugs, such as norepinephrine and L(G) mono-methyl-L-arginine, have harmful effects on the cerebral circulation. During experimental hyperdynamic sepsis, however, neither drug altered cerebral vascular resistance or cerebral blood flow. PMID- 12651672 TI - Splanchnic blood flow in low-flow states. AB - IMPLICATIONS: Insufficient splanchnic blood flow in critically ill patients is the result of a multitude of different diseases, treatment modalities and their interplay, and is associated with increased morbidity and mortality. A combination of diminished and heterogeneous mesenteric blood flow, impaired or exhausted regulatory mechanisms and adverse drug effects may coexist with normal systemic hemodynamics. PMID- 12651673 TI - The comparative effects of prostaglandin E1 and nicardipine on cerebral microcirculation in rabbits. AB - We compared the effects of the systemic hypotensive drugs prostaglandin E1 (PGE1) and nicardipine on the cerebral microcirculation and on the cerebrovascular reactivities to hypercapnia and hypoxia. In isoflurane-anesthetized rabbits (n = 48), we measured cerebral pial vessel diameters using a cranial-window preparation: (a) during IV PGE1- or nicardipine-induced mild or moderate hypotension (to 80% or 60% of initial mean arterial blood pressure), (b) after topical administration of these drugs, and (c) during hypercapnia or hypoxia induced during such mild or moderate hypotension. Pial arteriolar diameters were (a) unchanged when hypotension (mild or moderate) was induced by PGE1 but increased when it was induced by nicardipine and (b) increased dose-dependently by topical administration of nicardipine but not PGE1. Only small changes in cerebral venular diameter were observed in these experiments. The pial arteriolar dilator response to hypercapnia was potentiated during hypotension (mild or moderate) when it was induced by PGE1 but decreased when it was induced by nicardipine, whereas the response to hypoxia was maintained during PGE1-induced hypotension but decreased during nicardipine-induced hypotension. In conclusion, as a systemic hypotensive drug, PGE1 does not dilate cerebral arterioles and maintains cerebrovascular reactivities to hypercapnia and hypoxia, whereas nicardipine dilates such vessels and reduces these cerebrovascular reactivities. IMPLICATIONS: When given systemically to produce mild or moderate hypotension, prostaglandin E1 does not induce cerebral vasodilation and maintains cerebrovascular reactivity to hypercapnia and hypoxia, whereas nicardipine dilates cerebral vessels and reduces both reactivities. PMID- 12651674 TI - Esmolol blunts the cerebral blood flow velocity increase during emergence from anesthesia in neurosurgical patients. AB - Cerebral hyperemia has been demonstrated during emergence from anesthesia in neurosurgical patients, but its mechanism is speculative. We performed this study to test the hypothesis that this could be attributed to sympathetic overactivity. Thirty neurosurgical patients were included in a prospective, randomized, double blinded study comparing esmolol, a short-acting beta-blocker, and a placebo. Esmolol (0.3 mg. kg(-1). min(-1)) was infused from the end of anesthesia to 15 min after extubation. Cerebral blood flow velocity (CBFV), mean arterial blood pressure, and heart rate were recorded before anesthesia, during anesthesia after surgery, at extubation, and 5-60 min after extubation. Cardiac output (COe) was estimated by using an esophageal Doppler from anesthesia to 60 min after extubation. CBFV, COe, and heart rate were significantly lower in the esmolol group. Mean arterial blood pressure was comparable between the groups. There was no correlation between CBFV and COe at any time point during the study. In conclusion, esmolol blunted the CBFV increase during emergence, confirming that sympathetic overactivity contributes to cerebral hyperemia during neurosurgical recovery. IMPLICATIONS: Esmolol blunted the postoperative increase in cerebral blood flow velocity in neurosurgical patients. The origin of sympathetic hyperactivity and its potential deleterious consequences require further study. PMID- 12651675 TI - Penetration of intravenous hydroxyethyl starch into the cerebrospinal fluid in patients with impaired blood-brain barrier function. AB - Hypovolemic patients with impairment of the blood-brain barrier may receive IV hydroxyethyl starch (HES) to stabilize cardiovascular function and to increase cerebral perfusion pressure. It is not known whether HES can penetrate into the cerebrospinal fluid (CSF) under those conditions. We investigated plasma and CSF levels of HES after IV infusion in patients with suspected disturbance of the blood-brain barrier. Eight adult patients were studied who were being treated for head trauma or subarachnoid hemorrhage, with an external CSF drain in place. All patients exhibited radiographic signs of blood-brain barrier impairment diagnosed by cerebral computed tomography. After IV infusion of 500 to 1000 mL of HES 200,000/0.5, plasma HES levels were measured. Additionally, all CSF that was drained within 8 h after the HES infusion was collected, and HES concentrations were measured. All patients had detectable HES plasma concentrations (3.41 to 9.95 mg/mL). In contrast, no HES could be detected in the CSF of any patient. These data indicate that IV HES 200,000/0.5 does not penetrate into the CSF in patients with disturbed blood-brain barrier function after subarachnoid hemorrhage or head trauma. Further study is required to determine whether HES penetrates into the intracranial interstitium, despite the absence of HES in the CSF. IMPLICATIONS: Patients may receive IV hydroxyethyl starch (HES) after head trauma or subarachnoid hemorrhage. The results of the present study indicate that in patients with suspected blood-brain barrier impairment, HES does not penetrate from the plasma into the cerebrospinal fluid. PMID- 12651676 TI - Preservation of the cortical somatosensory-evoked potential during dexmedetomidine infusion in rats. AB - Successful somatosensory-evoked potential (SEP) monitoring has been performed during the administration of dexmedetomidine to patients, but a systematic investigation of the dose response of the SEP to dexmedetomidine has not been reported. In this study, we evaluated the effect of a range of dexmedetomidine doses on the cortical SEP in rats. Twelve rats were initially anesthetized with ketamine and the lungs were mechanically ventilated. Femoral arterial and venous catheters were placed. Anesthesia was maintained with constant infusions of remifentanil (5-15 microg. kg(-1). min(-1)) and vecuronium (56 microg. kg(-1). min(-1)). Dexmedetomidine was infused at 0.1, 0.25, 0.5, 1.0, and 2.0 microg. kg( 1). min(-1) in a stepwise manner with 10-min infusion periods at each step. In eight rats, an additional large-dose infusion of dexmedetomidine at 10 microg. kg(-1). min(-1) was administered for 30 min. The cortical SEPs were recorded after stimulation of the tibial nerve. At all infusion rates, there was a statistically insignificant increase in the SEP amplitude. Dexmedetomidine consistently increased the SEP latency, but these increases were not statistically significant. These data demonstrate that dexmedetomidine maintains technically adequate conditions for SEP monitoring in rats and provides support for future studies of the effect of dexmedetomidine on SEP monitoring in humans. IMPLICATIONS: In rats, the administration of a wide range of infusion rates of dexmedetomidine did not significantly affect the somatosensory-evoked potential. These results suggest that dexmedetomidine might be a useful adjunctive drug in patients undergoing intraoperative somatosensory-evoked potential monitoring. PMID- 12651677 TI - The effect of epidural neostigmine combined with ropivacaine and sufentanil on neuraxial analgesia during labor. AB - Spinal neostigmine produces analgesia without respiratory depression or hypotension but provokes major gastrointestinal side effects. Epidural injection of this drug, however, appears to induce analgesia devoid of such side effects. In this study, we evaluated the effect of a bolus of epidural neostigmine on the duration and magnitude of analgesia in early labor and assessed its eventual sparing effect on subsequent local anesthetic requirements. Epidural neostigmine methylsulfate (maximal dose 4 microg/kg) was added to 10 mL of ropivacaine 0.1%, with and without sufentanil 10 microg, to initiate analgesia. Twenty minutes after injection, pain score, sensory level, and motor block were assessed. Time until request for supplemental epidural medication was also recorded. Patient controlled epidural analgesia with ropivacaine 0.1% was used for epidural supplementation. Maternal and fetal side effects were closely recorded. Neostigmine (4 microg/kg), when added to ropivacaine 10 mg, provided equivalent analgesia to ropivacaine 20 mg but was less effective than sufentanil 10 microg for the initiation of labor epidural analgesia. Further, neostigmine did not modify the subsequent patient-controlled epidural analgesia local anesthetic requirements during labor. No hemodynamic instability, additional motor block, or bothersome side effects were recorded. IMPLICATIONS: The combination of epidural neostigmine (4 microg/kg) with the local anesthetic ropivacaine, with or without sufentanil, does not significantly enhance neuraxial analgesia during labor. Such a dose, however, has no bothersome side effects. PMID- 12651678 TI - The effect of epidural test dose on motor function after a combined spinal epidural technique for labor analgesia. AB - Labor analgesia initiated with intrathecal bupivacaine and fentanyl, without a local anesthetic epidural test dose, provides effective analgesia and allows ambulation. In this study, we sought to determine the effect of a lidocaine epinephrine test dose administered immediately after the initiation of combined spinal-epidural (CSE) analgesia with bupivacaine 2.5 mg and fentanyl 25 micro g on parturients' hemodynamic stability, posterior column function, motor strength, and subjective ability to walk. Parturients (n = 153) were randomized to receive either 3 mL of epidural saline or lidocaine 1.5% with epinephrine 1:200,000. Hemodynamic variables, proprioception, straight leg raise, and the modified Bromage score were analyzed in 110 parturients who completed the study protocol and were not different between groups. Vibratory sense, the ability to perform a partial deep knee bend and to step up on a stool, and the subjective ability to walk were impaired in a larger number of parturients in the lidocaine-epinephrine group at 30 min (P < 0.05). At 60 min, there were no differences between the groups except that fewer parturients in the lidocaine-epinephrine group could step up on a stool. The straight leg raise against resistance and the modified Bromage scale did not correlate well with other tests of motor strength (Spearman's rho, 0.273-0.405). These data suggest that the test dose should be avoided immediately after initiation of CSE analgesia when early ambulation is desired. IMPLICATIONS: A lidocaine-epinephrine epidural test dose (3 mL of lidocaine 1.5% with epinephrine 1:200,000), injected immediately after the initiation of combined spinal-epidural labor analgesia with bupivacaine 2.5 mg and fentanyl 25 microg, may interfere with the ability to perform simple tests of motor function and ambulation. PMID- 12651679 TI - Ropivacaine 0.15% plus sufentanil 0.5 microg/mL and ropivacaine 0.10% plus sufentanil 0.5 microg/mL are equivalent for patient-controlled epidural analgesia during labor. AB - We compared the administration of 0.15% ropivacaine plus 0.5 microg/mL of sufentanil with that of 0.10% ropivacaine plus 0.5 microg/mL of sufentanil for labor analgesia with patient-controlled epidural analgesia (PCEA) to determine whether a decreased concentration of ropivacaine could produce equally effective analgesia. One-hundred-thirty healthy pregnant women at term were randomized in a double-blinded fashion. The PCEA settings were as follows: 12-mL initial bolus, 5 mL bolus dose, 5-min lockout interval, and 10 mL/h basal infusion. Patient demographics and labor characteristics were comparable in both groups. No differences were observed for pain scores, maternal satisfaction, volume of anesthetic solution administered, number of boluses requested and delivered, need for supplemental boluses, mode of delivery, motor block, side effects, or Apgar scores. Patients in the 0.10% ropivacaine group used significantly less drug than those in the 0.15% group (mean, 57 mg; 95% confidence interval, 50.5-63.5 mg; versus mean, 88.0 mg; 95% confidence interval, 74.4-93.3 mg, respectively; P < 0.0001). Ropivacaine 0.10% plus 0.5 microg/mL of sufentanil administered via PCEA for labor analgesia is equally effective as ropivacaine 0.15% plus 0.5 microg/mL of sufentanil, with a 30% local anesthetic-sparing effect and a 40% reduction in cost. However, this reduction in local anesthetic is not associated with a decrease in the incidence of motor block, side effects, or instrumental deliveries. IMPLICATIONS: Ropivacaine 0.10% plus 0.5 microg/mL of sufentanil given via patient-controlled epidural anesthesia for labor analgesia is equally as effective as ropivacaine 0.15% plus 0.5 microg/mL of sufentanil, with a 30% local anesthetic-sparing effect and a 40% reduction in cost. This reduction in ropivacaine concentration is not associated with a decrease in the incidence of motor block, side effects, or instrumental deliveries. PMID- 12651680 TI - Minimum analgesic doses of fentanyl and sufentanil for epidural analgesia in the first stage of labor. AB - In this study, we sought to determine the minimum analgesic doses and relative potencies of fentanyl and sufentanil when they are used as the sole epidural analgesic during the first stage of labor. Nulliparous parturients (n = 66) in spontaneous labor at term gestation and requesting epidural analgesia were enrolled into this prospective, double-blinded, randomized, sequential-allocation study. Each woman received fentanyl or sufentanil diluted with 0.9% wt/vol saline to a volume of 10 mL. The initial dose was arbitrarily chosen to be 125 microg for fentanyl and 25 microg for sufentanil, with subsequent doses being determined by the response of the previous patient (testing interval, 5 microg for fentanyl and 1 microg for sufentanil). Efficacy was accepted if the visual analog score decreased to < or =10 mm on a 100-mm scale within 30 min. The minimum analgesic dose or median effective dose was 21.1 microg (95% confidence interval [CI], 20.2 21.9 microg) for sufentanil and 124.2 microg (95% CI, 118.1-130.6 microg) for fentanyl (P < 0.0001). The sufentanil/fentanyl potency ratio was 5.9 (95% CI, 5.6 6.3). In conclusion, we have established the equivalent doses and relative potencies of fentanyl and sufentanil for epidural analgesia in the first stage of labor. IMPLICATIONS: This study determined the minimum analgesic doses of fentanyl and sufentanil for epidural anesthesia in the first stage of labor. The sufentanil/fentanyl potency ratio was 5.9. This ratio may be used to establish the equivalent doses for fentanyl and sufentanil for epidural analgesia in labor. PMID- 12651681 TI - Clinical results with the acoustic puncture assist device, a new acoustic device to identify the epidural space. AB - Sixty patients scheduled for lumbar epidural anesthesia were included in a study in which we evaluated the efficacy of localizing the epidural space by means of an acoustic signal. A prototype of an acoustic puncture assist device, connected to the epidural needle by an extension tube, generated the pressure needed to perform the epidural puncture and translated this pressure into corresponding acoustic and visible signals. The device frees the anesthesiologist to handle the epidural needle with both hands and to detect the epidural space by means of these signals. In all 60 patients (100%), the epidural space was successfully located by using the acoustic signal. In all cases, this was confirmed by the pressure measurement, which proved to be a reliable indicator for correct identification of the epidural space. We conclude that it is possible to locate the epidural space by means of the acoustic puncture assist device. The method proved to be reliable, safe, and simple in this study. The benefits of this new epidural puncture technique include better needle control, teaching, control of correct catheter placement, and documentation. The last can be an important adjunct to anesthesia practice. IMPLICATIONS: The authors demonstrate that it is possible to identify the epidural space by an acoustic and visible signal. An experimental setup constructed for this purpose makes the epidural puncture procedure audible and visible. PMID- 12651682 TI - Seizure after levobupivacaine for interscalene brachial plexus block. AB - IMPLICATIONS: This case report describes a patient who demonstrated generalized seizure activity after an injection of 30 mL of levobupivacaine 0.5% for interscalene brachial plexus block. No evidence of cardiovascular toxicity was noted. PMID- 12651683 TI - Airway difficulty after a brachial plexus subclavian perivascular block. AB - IMPLICATIONS: We report a case of upper airway obstruction after subclavian perivascular block in a patient with a preexisting unrecognized paralyzed vocal cord on the opposite side. We discuss the incidence of vocal cord paralysis after subclavian perivascular block and the risk of airway compromise if contralateral vocal cord paralysis is known or suspected. PMID- 12651684 TI - From Victor Pauchet to Gaston Labat: the transformation of regional anesthesia from a surgeon's practice to the physician anesthesiologist. PMID- 12651686 TI - The influence of induced hypothermia for hemostatic function on temperature adjusted measurements in rabbits. AB - In hypothermic patients, a tendency to bleed may be observed even when hemostatic tests seem to be normal. Coagulation and platelet function tests are usually performed at 37 degrees C. We investigated the influence of induced hypothermia on temperature-adjusted hemostasis function testing using Sonoclot Analyzer (Sonoclot) and Thromboelastography (TEG). Anesthesia was induced and maintained with IV ketamine and fentanyl on 15 male New-Zealand White rabbits. A water blanket was used to induce hypothermia to 30 degrees C and to rewarm to 37 degrees C. Blood samples were obtained at four points: before hypothermia, at 34 degrees C, at 30 degrees C, and after rewarming. Standard coagulation tests were performed at 37 degrees C (C method), and simultaneously, real temperature hemostasis function tests (R method) were run. In Sonoclot(R), activated clotting time and time to peak increased and clot rate decreased significantly at 30 degrees C in the R method compared with those in the C method. In TEG(R), reaction time and clot formation time were prolonged and clot formation rate was diminished at 30 degrees C in the R method compared with those in the C method. Induced hypothermia delayed the coagulation cascade and reduced platelet function. During hypothermia, hemostatic measurements should be performed at real temperature to avoid overestimating patient hemostatic function based on results measured at the standard 37 degrees C. IMPLICATIONS: We investigated the influence of induced hypothermia on temperature-adjusted hemostasis function tests in rabbits using Sonoclot Analyzer and Thromboelastography. Induced hypothermia delayed the coagulation cascade and reduced platelet function. The conventional coagulation tests performed at 37 degrees C failed to detect these hypothermia-induced degradations in hemostasis performance. PMID- 12651685 TI - Treating intraoperative hyperchloremic acidosis with sodium bicarbonate or tris hydroxymethyl aminomethane: a randomized prospective study. AB - In this study, we evaluated the action of two buffer solutions on acid-base equilibrium in cases of hyperchloremic acidosis. Twenty-four patients undergoing major gynecological intraabdominal surgery received 40 mL. kg(-1). h(-1) of 0.9% saline per protocol. During surgery, in every patient, hyperchloremic acidosis occurred. At a standard base excess of -7 mmol/L, the patients were randomly assigned to receive within 20 min either a mean of 130 +/- 26 mmol of sodium bicarbonate (BIC, 1 M; n = 12) or a mean of 128 +/- 18 mmol of tris-hydroxymethyl aminomethane (THAM, 3 M; n = 12). PaCO(2), pH, serum bicarbonate concentration, standard base excess, and serum concentrations of sodium, potassium, chloride, lactate, phosphate, total protein, and albumin were determined before and 0, 10, and 20 min after buffering. The apparent strong ion difference was calculated as: serum sodium plus serum potassium minus serum chloride minus serum lactate. The effective strong ion difference and the amount of weak plasma acid were calculated by using a computer program. Immediately after buffering, standard base excess increased by 9.8 mmol/L in the BIC group and by 7.2 mmol/L in the THAM group. In both groups, PaCO(2) and the amount of weak plasma acid remained constant. Mainly because of hypernatremia, the apparent and effective strong ion difference increased in the BIC group by 8.5 and 7.9 mEq/L, respectively. In the THAM group, the apparent strong ion difference remained constant; however, the effective strong ion difference increased by 6.4 mEq/L and the anion gap decreased by 5.8 mmol/L because of the occurrence of an unmeasured cation. In conclusion, in case of buffering with BIC or THAM, the changes in pH were accompanied by, and probably caused by, an increase in strong ion difference. IMPLICATIONS: By comparing two groups of patients with intraoperative hyperchloremic acidosis receiving equal doses of either sodium bicarbonate or tris-hydroxymethyl aminomethane, we assessed the action of both drugs on acid base equilibrium. In case of "buffering," the changes in pH were accompanied by, and probably caused by, an increase in strong ion difference. PMID- 12651687 TI - A comparison of the PAxpress and face mask plus Guedel airway by inexperienced personnel after mannequin-only training. AB - The PAxpress (PAX) is a new extraglottic airway device consisting of an anatomically curved tube, an inflatable circular cuff in the midsection, a noninflatable gilled conical cuff at the distal end, and an anterior-facing, rectangular hooded vent between the two cuffs. We compared the ability of nurses with no previous airway management experience to ventilate the lungs of 45 ASA physical status I and II anesthetized, paralyzed patients using either the PAX or face mask and Guedel airway (FM/GA) after mannequin-only training. Nurses were asked to ventilate the patient to an expired target tidal volume of 7 mL/kg and then to the maximum tidal volume achievable. The FM/GA was used first and the then the PAX. The target tidal volume was achieved in all patients with both devices. There were no differences in the time taken (PAX, 41 +/- 15 s; FM/GA, 39 +/- 25 s) or the number of insertion attempts to achieve the target tidal volume. There were no differences in the frequency of esophageal leaks at the target (PAX, 9%; FM/GA, 4%) and maximum tidal volume (PAX, 51%; FM/GA, 49%). The maximal tidal volume (1261 +/- 306 versus 958 +/- 220 mL; P < 0.0001) and peak airway pressure (37 +/- 5 versus 28 +/- 6 cm H(2)O; P < 0.0001) was larger for the PAX, but blood was detected more frequently (22% versus 0%; P = 0.001). We conclude that ventilation to a target tidal volume of 7 mL/kg in anesthetized, paralyzed adults is equally successful for the PAX and FM/GA by inexperienced nurses after mannequin-only training. However, the maximal tidal volume and peak airway pressure is larger and airway trauma more common with the PAX. IMPLICATIONS: Ventilation to a target tidal volume of 7 mL/kg in anesthetized, paralyzed adults is equally successful for the PAxpress (PAX) and Face Mask/Guedel Airway by inexperienced nurses after mannequin-only training. However, the maximal tidal volume and peak airway pressure is larger and airway trauma more common with the PAX. PMID- 12651688 TI - Is peripheral line placement more difficult in obese than in lean patients? AB - IMPLICATIONS: This work shows that the insertion of a peripheral venous line is more difficult in obese than in lean patients. The usual recommendation, that a central venous line should be inserted routinely in obese patients to perform anesthesia, is not valid. PMID- 12651689 TI - Latex allergy: an update. PMID- 12651690 TI - Fibrinolysis during liver transplant and use of solvent/detergent virus inactivated plasma (ESDEP/Octaplas). PMID- 12651691 TI - False increase BIS values with forced-air head warming. PMID- 12651692 TI - Liver transplantation, solvent-detergent treated plasma and antifibrinolytics. PMID- 12651693 TI - Artifactual dextrocardia associated with a defect in the ECG labeling. PMID- 12651694 TI - Massive hemorrhage during radiofrequency ablation of a pulmonary neoplasm. PMID- 12651695 TI - The difficult airway algorithm of the American Society of Anesthesiologists. PMID- 12651696 TI - Density determination of bupivacaine and bupivacaine-opioid mixtures for spinal anesthesia. PMID- 12651697 TI - Entrapment of an exchange wire by an inferior vena caval filter: a technique for removal. PMID- 12651698 TI - COX-2 inhibition for postoperative analgesia. PMID- 12651699 TI - Temperature measurements. PMID- 12651700 TI - Comparison of computer based and patient simulator based crisis. PMID- 12651701 TI - Virginia Mason Medical Center's promotion of outpatient lidocaine spinal anesthesia questioned. PMID- 12651702 TI - Right subclavian artery injury. PMID- 12651703 TI - A warm bed. PMID- 12651704 TI - Ketamine for restless legs syndrome. PMID- 12651705 TI - Discoloration of parenteral ondansetron. PMID- 12651706 TI - Is fast induction with sevoflurane associated with an increased anesthetic risk in pediatric patients? PMID- 12651707 TI - Possible right lung isolation by blocking the tracheal bronchus with only a Univent tube for some patients. PMID- 12651708 TI - Improving pulse oximetry signal quality. PMID- 12651709 TI - Insertion of a laryngeal mask airway in the prone position. PMID- 12651710 TI - Indirect intubation. PMID- 12651711 TI - Triplet repeats in human genome: distribution and their association with genes and other genomic regions. AB - MOTIVATION: Simple sequence repeats (SSRs) or microsatellite repeats are found abundantly in many prokaryotic and eukaryotic genomes. Among SSRs, triplet repeats are of special significance because some of them have been linked to various genetic disorders. The objective of the study is to analyze the triplet repeats of complete human genome and to identify the genes that contain the triplet repeats in their coding region. The analysis will help us to identify the candidate genes that have potential for repeat expansion. RESULTS: We have analyzed triplet repeats in the complete human genome from the publicly available sequences. Our analysis revealed that AGC and CCG repeat were predominantly present in the coding regions of the genome while UTRs and the upstream sequences contained CCG repeats in relative abundance. Analysis of density of triplet repeats (bp/Mb) revealed that AAT and AAC were the abundant repeats whereas ACT and ACG were the rare repeats found in human genome. We could identify about 2135 known or predicted genes that were associated with at least one of the triplet repeat types. A large proportion of putative transcripts that were identified by gene finding programs were found to be associated with triplet repeats. These transcripts will be the candidate genes for analysis of triplet repeat expansion and a possible association with disease phenotypes. Identification of 171 genes which contain a minimum of ten repeat units will be of particular interest in future in correlating their association with any disease phenotype due to the expansion potential of repeats present in them. The list of genes and other details of analysis are given in the online supplementary data (http://www.ingenovis.com/tripletrepeats). PMID- 12651712 TI - Automatic analysis of DNA microarray images using mathematical morphology. AB - MOTIVATION: DNA microarrays are an experimental technology which consists in arrays of thousands of discrete DNA sequences that are printed on glass microscope slides. Image analysis is an important aspect of microarray experiments. The aim of this step is to reduce an image of spots into a table with a measure of the intensity for each spot. Efficient, accurate and automatic analysis of DNA spot images is essential in order to use this technology in laboratory routines. RESULTS: We present an automatic non-supervised set of algorithms for a fast and accurate spot data extraction from DNA microarrays using morphological operators which are robust to both intensity variation and artefacts. The approach can be summarised as follows. Initially, a gridding algorithm yields the automatic segmentation of the microarray image into spot quadrants which are later individually analysed. Then the analysis of the spot quadrant images is achieved in five steps. First, a pre-quantification, the spot size distribution law is calculated. Second, the background noise extraction is performed using a morphological filtering by area. Third, an orthogonal grid provides the first approach to the spot locus. Fourth, the spot segmentation or spot boundaries definition is carried out using the watershed transformation. And fifth, the outline of detected spots allows the signal quantification or spot intensities extraction; in this respect, a noise model has been investigated. The performance of the algorithm has been compared with two packages: ScanAlyze and Genepix, showing its robustness and precision. PMID- 12651713 TI - Effective dimension reduction methods for tumor classification using gene expression data. AB - MOTIVATION: One particular application of microarray data, is to uncover the molecular variation among cancers. One feature of microarray studies is the fact that the number n of samples collected is relatively small compared to the number p of genes per sample which are usually in the thousands. In statistical terms this very large number of predictors compared to a small number of samples or observations makes the classification problem difficult. An efficient way to solve this problem is by using dimension reduction statistical techniques in conjunction with nonparametric discriminant procedures. RESULTS: We view the classification problem as a regression problem with few observations and many predictor variables. We use an adaptive dimension reduction method for generalized semi-parametric regression models that allows us to solve the 'curse of dimensionality problem' arising in the context of expression data. The predictive performance of the resulting classification rule is illustrated on two well know data sets in the microarray literature: the leukemia data that is known to contain classes that are easy 'separable' and the colon data set. PMID- 12651714 TI - PCA disjoint models for multiclass cancer analysis using gene expression data. AB - MOTIVATION: Microarray expression profiling appears particularly promising for a deeper understanding of cancer biology and to identify molecular signatures supporting the histological classification schemes of neoplastic specimens. However, molecular diagnostics based on microarray data presents major challenges due to the overwhelming number of variables and the complex, multiclass nature of tumor samples. Thus, the development of marker selection methods, that allow the identification of those genes that are most likely to confer high classification accuracy of multiple tumor types, and of multiclass classification schemes is of paramount importance. RESULTS: A computational procedure for marker identification and for classification of multiclass gene expression data through the application of disjoint principal component models is described. The identified features represent a rational and dimensionally reduced base for understanding the basic biology of diseases, defining targets for therapeutic intervention, and developing diagnostic tools for the identification and classification of multiple pathological states. The method has been tested on different microarray data sets obtained from various human tumor samples. The results demonstrate that this procedure allows the identification of specific phenotype markers and can classify previously unseen instances in the presence of multiple classes. PMID- 12651715 TI - Mapping multiple co-sequenced T-DNA integration sites within the Arabidopsis genome. AB - MOTIVATION: Insertion mutagenesis, using transgenes or endogenous transposons, is a popular method for generating null mutations (knockouts) in model organisms. Insertions are mapped to specific genes by amplifying (via TAIL-PCR) and sequencing genomic regions flanking the inserted DNA. The presence of multiple TAIL-PCR templates in one sequencing reaction results in chimeric sequence of intermittently low quality. Standard processing of this sequence by applying Phred quality requirements results in loss of informative sequence, whereas not trimming low-quality sequence causes inclusion of low-complexity homopolymers from the ends of sequence runs. Accurate mapping of the flanking sequences is complicated by the presence of gene families. RESULTS: Methods for extracting informative regions from sequence traces obtained by sequencing multiple TAIL-PCR fragments in a single reaction are described. The completely sequenced Arabidopsis genome was used to identify informative TAIL-PCR sequence regions. Methods were devised to define and select high quality matches and precisely map each insert to the correct genome location. These methods were used to analyze sequence of TAIL-PCR-amplified flanking regions of the inserts from individual plants in a T-DNA-mutagenized population of Arabidopsis thaliana, and are applicable to similar situations where a reference genome can be used to extract information from poor-quality sequence. PMID- 12651716 TI - The fuzzy polynucleotide space: basic properties. AB - MOTIVATION: Any triplet codon may be regarded as a 12-dimensional fuzzy code. Sufficient information about a particular sequence may not be available in certain situations. The investigator will be confronted with imprecise sequences, yet want to make comparisons of sequences. Fuzzy polynucleotides can be compared by using geometrical interpretation of fuzzy sets as points in a hypercube. RESULTS: We introduce the space of fuzzy polynucleotides and a means of measuring dissimilitudes between them. We establish mathematical principles to measure dissimilarities between fuzzy polynucleotides and present several examples in this metric space. We calculate the frequencies of the nucleotides at the three base sites of a codon in the coding sequences of Escherichia coli K-12 and Mycobacterium tuberculosis H37Rv, and consider them as points in that fuzzy space. We compute the distance between the genomes of E.coli and M.tuberculosis. PMID- 12651717 TI - The Z curve database: a graphic representation of genome sequences. AB - MOTIVATION: Genome projects for many prokaryotic and eukaryotic species have been completed and more new genome projects are being underway currently. The availability of a large number of genomic sequences for researchers creates a need to find graphic tools to study genomes in a perceivable form. The Z curve is one of such tools available for visualizing genomes. The Z curve is a unique three-dimensional curve representation for a given DNA sequence in the sense that each can be uniquely reconstructed given the other. The Z curve database for more than 1000 genomes have been established here. RESULTS: The database contains the Z curves for archaea, bacteria, eukaryota, organelles, phages, plasmids, viroids and viruses, whose genomic sequences are currently available. All the 3 dimensional Z curves and their three component curves are stored in the database. The applications of the Z curve database on comparative genomics, gene prediction, computation of G+C content with a windowless technique, prediction of replication origins and terminations of bacterial and archaeal genomes and study of local deviations from the Chargaff Parity Rule 2 etc. are presented in detail. The Z curve database reported here is a treasure trove in which biologists could find useful biological knowledge. PMID- 12651718 TI - The estimation of relative site variability among aligned homologous protein sequences. AB - MOTIVATION: Maximum likelihood-based methods to estimate site by site substitution rate variability in aligned homologous protein sequences rely on the formulation of a phylogenetic tree and generally assume that the patterns of relative variability follow a pre-determined distribution. We present a phylogenetic tree-independent method to estimate the relative variability of individual sites within large datasets of homologous protein sequences. It is based upon two simple assumptions. Firstly that substitutions observed between two closely related sequences are likely, in general, to occur at the most variable sites. Secondly that non-conservative amino acid substitutions tend to occur at more variable sites. Our methodology makes no assumptions regarding the underlying pattern of relative variability between sites. RESULTS: We have compared, using data simulated under a non-gamma distributed model, the performance of this approach to that of a maximum likelihood method that assumes gamma distributed rates. At low mean rates of evolution our method inferred site by site relative substitution rates more accurately than the maximum likelihood approach in the absence of prior assumptions about the relationships between sequences. Our method does not directly account for the effects of mutational saturation, However, we have incorporated an 'ad-hoc' modification that allows the accurate estimation of relative site variability in fast evolving and saturated datasets. PMID- 12651719 TI - Greedy mixture learning for multiple motif discovery in biological sequences. AB - MOTIVATION: This paper studies the problem of discovering subsequences, known as motifs, that are common to a given collection of related biosequences, by proposing a greedy algorithm for learning a mixture of motifs model through likelihood maximization. The approach adds sequentially a new motif to a mixture model by performing a combined scheme of global and local search for appropriately initializing its parameters. In addition, a hierarchical partitioning scheme based on kd-trees is presented for partitioning the input dataset in order to speed-up the global searching procedure. The proposed method compares favorably over the well-known MEME approach and treats successfully several drawbacks of MEME. RESULTS: Experimental results indicate that the algorithm is advantageous in identifying larger groups of motifs characteristic of biological families with significant conservation. In addition, it offers better diagnostic capabilities by building more powerful statistical motif-models with improved classification accuracy. PMID- 12651720 TI - How Bayes tests of molecular phylogenies compare with frequentist approaches. AB - MOTIVATION: The desire to compare molecular phylogenies has stimulated the design of numerous tests. Most of these tests are formulated in a frequentist framework, and it is not known how they compare with Bayes procedures. I propose here two new Bayes tests that either compare pairs of trees (Bayes hypothesis test, BHT), or test each tree against an average of the trees included in the analysis (Bayes significance test, BST). RESULTS: The algorithm, based on a standard Metropolis Hastings sampler, integrates nuisance parameters out and estimates the probability of the data under each topology. These quantities are used to estimate Bayes factors for composite vs. composite hypotheses. Based on two data sets, the BHT and BST are shown to construct similar confidence sets to the bootstrap and the Shimodaira Hasegawa test, respectively. This suggests that the known difference among previous tests is mainly due to the null hypothesis considered. PMID- 12651721 TI - MINRMS: an efficient algorithm for determining protein structure similarity using root-mean-squared-distance. AB - MOTIVATION: Existing algorithms for automated protein structure alignment generate contradictory results and are difficult to interpret. An algorithm which can provide a context for interpreting the alignment and uses a simple method to characterize protein structure similarity is needed. RESULTS: We describe a heuristic for limiting the search space for structure alignment comparisons between two proteins, and an algorithm for finding minimal root-mean-squared distance (RMSD) alignments as a function of the number of matching residue pairs within this limited search space. Our alignment algorithm uses coordinates of alpha-carbon atoms to represent each amino acid residue and requires a total computation time of O(m(3) n(2)), where m and n denote the lengths of the protein sequences. This makes our method fast enough for comparisons of moderate-size proteins (fewer than approximately 800 residues) on current workstation-class computers and therefore addresses the need for a systematic analysis of multiple plausible shape similarities between two proteins using a widely accepted comparison metric. PMID- 12651722 TI - Prediction of human protein function according to Gene Ontology categories. AB - MOTIVATION: The human genome project has led to the discovery of many human protein coding genes which were previously unknown. As a large fraction of these are functionally uncharacterized, it is of interest to develop methods for predicting their molecular function from sequence. RESULTS: We have developed a method for prediction of protein function for a subset of classes from the Gene Ontology classification scheme. This subset includes several pharmaceutically interesting categories-transcription factors, receptors, ion channels, stress and immune response proteins, hormones and growth factors can all be predicted. Although the method relies on protein sequences as the sole input, it does not rely on sequence similarity, but instead on sequence derived protein features such as predicted post translational modifications (PTMs), protein sorting signals and physical/chemical properties calculated from the amino acid composition. This allows for prediction of the function for orphan proteins where no homologs can be found. Using this method we propose two novel receptors in the human genome, and further demonstrate chromosomal clustering of related proteins. PMID- 12651723 TI - Dynamic modeling of genetic networks using genetic algorithm and S-system. AB - MOTIVATION: The modeling of system dynamics of genetic networks, metabolic networks or signal transduction cascades from time-course data is formulated as a reverse-problem. Previous studies focused on the estimation of only network structures, and they were ineffective in inferring a network structure with feedback loops. We previously proposed a method to predict not only the network structure but also its dynamics using a Genetic Algorithm (GA) and an S-system formalism. However, it could predict only a small number of parameters and could rarely obtain essential structures. In this work, we propose a unified extension of the basic method. Notable improvements are as follows: (1) an additional term in its evaluation function that aims at eliminating futile parameters; (2) a crossover method called Simplex Crossover (SPX) to improve its optimization ability; and (3) a gradual optimization strategy to increase the number of predictable parameters. RESULTS: The proposed method is implemented as a C program called PEACE1 (Predictor by Evolutionary Algorithms and Canonical Equations 1). Its performance was compared with the basic method. The comparison showed that: (1) the convergence rate increased about 5-fold; (2) the optimization speed was raised about 1.5-fold; and (3) the number of predictable parameters was increased about 5-fold. Moreover, we successfully inferred the dynamics of a small genetic network constructed with 60 parameters for 5 network variables and feedback loops using only time-course data of gene expression. PMID- 12651724 TI - TIGR Gene Indices clustering tools (TGICL): a software system for fast clustering of large EST datasets. AB - TGICL is a pipeline for analysis of large Expressed Sequence Tags (EST) and mRNA databases in which the sequences are first clustered based on pairwise sequence similarity, and then assembled by individual clusters (optionally with quality values) to produce longer, more complete consensus sequences. The system can run on multi-CPU architectures including SMP and PVM. PMID- 12651725 TI - DigiNorthern, digital expression analysis of query genes based on ESTs. AB - DigiNorthern (DN) is a web-based tool for virtually displaying expression profiles of query genes based on EST sequences. Two utilities are available: DN1 takes one query gene and quantitatively display its expression levels in tissues/organs that express the gene with comparison between normal and neoplastic status of each tissue; DN2 takes two sequences as query genes and compares their expression profiles side by side. PMID- 12651726 TI - Gene expression data preprocessing. AB - We present an interactive web tool for preprocessing microarray gene expression data. It analyses the data, suggests the most appropriate transformations and proceeds with them after user agreement. The normal preprocessing steps include scale transformations, management of missing values, replicate handling, flat pattern filtering and pattern standardization and they are required before performing any pattern analysis. The processed data set can be sent to other pattern analysis tools. PMID- 12651728 TI - MatArray: a Matlab toolbox for microarray data. AB - The microarray technology allows the high-throughput quantification of the mRNA level of thousands of genes under dozens of conditions, generating a wealth of data which must be analyzed using some form of computational means. A popular framework for such analysis is Matlab, a powerful computing language for which many functions have been written. However, although complex topics like neural networks or principal component analysis are freely available in Matlab, functions to perform more basic tasks like data normalization or hierarchical clustering in an efficient manner are not. The MatArray toolbox aims at filling this gap by offering efficient implementations of the most needed functions for microarray analysis. The functions in the toolbox are command-line only, since it is geared toward seasoned Matlab users. PMID- 12651727 TI - Engene: the processing and exploratory analysis of gene expression data. AB - Engene is a versatile, and platform-independent web tool for exploratory analysis of gene expression data that aims at storing, visualizing and processing large sets of gene expression patterns. PMID- 12651729 TI - TRAIT (TRAnscript Integrated Table): a knowledgebase of human skeletal muscle transcripts. AB - TRAIT is a knowledgebase integrating information on transcripts with related data from genome, proteins, ortholog genes and diseases. It was initially built as a system to manage an EST-based gene discovery project on human skeletal muscle, which yielded over 4500 independent sequence clusters. Transcripts are annotated using automatic as well as manual procedures, linking known transcripts to public databases and unknown transcripts to tables of predicted features. Data are stored in a MySQL database. Complex queries are automatically built by means of a user-friendly web interface that allows the concurrent selection of many fields such as ontology, expression level, map position and protein domains. The results are parsed by the system and returned in a ranked order, in respect to the number of satisfied criteria. PMID- 12651730 TI - MENDB: a database of polymorphic loci from natural populations. AB - MENDB is an online database for genetic markers determined to be polymorphic in natural populations. The database contains primer sequences and conditions for PCR, taxonomic information, and links to GenBank records, as well as basic statistics on the level of polymorphism for the surveyed populations/individuals. PMID- 12651731 TI - MHCBN: a comprehensive database of MHC binding and non-binding peptides. AB - MHCBN is a comprehensive database of Major Histocompatibility Complex (MHC) binding and non-binding peptides compiled from published literature and existing databases. The latest version of the database has 19 777 entries including 17 129 MHC binders and 2648 MHC non-binders for more than 400 MHC molecules. The database has sequence and structure data of (a) source proteins of peptides and (b) MHC molecules. MHCBN has a number of web tools that include: (i) mapping of peptide on query sequence; (ii) search on any field; (iii) creation of data sets; and (iv) online data submission. The database also provides hypertext links to major databases like SWISS-PROT, PDB, IMGT/HLA-DB, GenBank and PUBMED. PMID- 12651732 TI - GPCR-GRAPA-LIB--a refined library of hidden Markov Models for annotating GPCRs. AB - GPCR-GRAPA-LIB is a library of HMMs describing G protein coupled receptor families. These families are initially defined by class of receptor ligand, with divergent families divided into subfamilies using phylogenic analysis and knowledge of GPCR function. Protein sequences are applied to the models with the GRAPA curve-based selection criteria. RefSeq sequences for Homo sapiens, Drosophila melanogaster, and Caenorhabditis elegans have been annotated using this approach. PMID- 12651733 TI - PedNavigator: a pedigree drawing servlet for large and inbred populations. AB - PedNavigator is a pedigree drawing application for large and complex pedigrees. It has been developed especially for genetic and epidemiological studies of isolated populations characterized by high inbreeding and multiple matrimonies. PedNavigator is written in Java and is intended as a server-side web application, allowing researchers to 'walk' through family ties by point-and-clicking on person's symbols. The application is able to enrich the pedigree drawings with genotypic and phenotypic information taken from the underlying relational database. PMID- 12651734 TI - TreeSAAP: selection on amino acid properties using phylogenetic trees. AB - The software program TreeSAAP measures the selective influences on 31 structural and biochemical amino acid properties during cladogenesis, and performs goodness of-fit and categorical statistical tests. PMID- 12651735 TI - DomCut: prediction of inter-domain linker regions in amino acid sequences. AB - DomCut is a program to predict inter-domain linker regions solely by amino acid sequence information. The prediction is made by using linker index deduced from a data set of domain/linker segments. The linker preference profile, which is the averaged linker index along a sequence, can be visualized in the graphical interface. PMID- 12651736 TI - ASAP: automated sequence annotation pipeline for web-based updating of sequence information with a local dynamic database. AB - The automated sequence annotation pipeline (ASAP) is designed to ease routine investigation of new functional annotations on unknown sequences, such as expressed sequence tags (ESTs), through querying of web-accessible resources and maintenance of a local database. The system allows easy use of the output from one search as the input for a new search, as well as the filtering of results. The database is used to store formats and parameters and information for parsing data from web sites. The database permits easy updating of format information should a site modify the format of a query or of a returned web page. PMID- 12651737 TI - Cellerator: extending a computer algebra system to include biochemical arrows for signal transduction simulations. AB - Cellerator describes single and multi-cellular signal transduction networks (STN) with a compact, optionally palette-driven, arrow-based notation to represent biochemical reactions and transcriptional activation. Multi-compartment systems are represented as graphs with STNs embedded in each node. Interactions include mass-action, enzymatic, allosteric and connectionist models. Reactions are translated into differential equations and can be solved numerically to generate predictive time courses or output as systems of equations that can be read by other programs. Cellerator simulations are fully extensible and portable to any operating system that supports Mathematica, and can be indefinitely nested within larger data structures to produce highly scaleable models. PMID- 12651738 TI - Mass transport in arteries and the localization of atherosclerosis. AB - Atherosclerosis is a disease of the large arteries that involves a characteristic accumulation of high-molecular-weight lipoprotein in the arterial wall. This review focuses on the mass transport processes that mediate the focal accumulation of lipid in arteries and places particular emphasis on the role of fluid mechanical forces in modulating mass transport phenomena. In the final analysis, four mass transport mechanisms emerge that may be important in the localization of atherosclerosis: blood phase controlled hypoxia, leaky endothelial junctions, transient intercellular junction remodeling, and convective clearance of the subendothelial intima and media. Further study of these mechanisms may contribute to the development of therapeutic strategies for atherosclerotic diseases. PMID- 12651739 TI - The RNA polymerase II core promoter. AB - The events leading to transcription of eukaryotic protein-coding genes culminate in the positioning of RNA polymerase II at the correct initiation site. The core promoter, which can extend ~35 bp upstream and/or downstream of this site, plays a central role in regulating initiation. Specific DNA elements within the core promoter bind the factors that nucleate the assembly of a functional preinitiation complex and integrate stimulatory and repressive signals from factors bound at distal sites. Although core promoter structure was originally thought to be invariant, a remarkable degree of diversity has become apparent. This article reviews the structural and functional diversity of the RNA polymerase II core promoter. PMID- 12651740 TI - Signals for sorting of transmembrane proteins to endosomes and lysosomes. AB - Sorting of transmembrane proteins to endosomes and lysosomes is mediated by signals present within the cytosolic domains of the proteins. Most signals consist of short, linear sequences of amino acid residues. Some signals are referred to as tyrosine-based sorting signals and conform to the NPXY or YXXO consensus motifs. Other signals known as dileucine-based signals fit [DE]XXXL[LI] or DXXLL consensus motifs. All of these signals are recognized by components of protein coats peripherally associated with the cytosolic face of membranes. YXXO and [DE]XXXL[LI] signals are recognized with characteristic fine specificity by the adaptor protein (AP) complexes AP-1, AP-2, AP-3, and AP-4, whereas DXXLL signals are recognized by another family of adaptors known as GGAs. Several proteins, including clathrin, AP-2, and Dab2, have been proposed to function as recognition proteins for NPXY signals. YXXO and DXXLL signals bind in an extended conformation to the mu2 subunit of AP-2 and the VHS domain of the GGAs, respectively. Phosphorylation events regulate signal recognition. In addition to peptide motifs, ubiquitination of cytosolic lysine residues also serves as a signal for sorting at various stages of the endosomal-lysosomal system. Conjugated ubiquitin is recognized by UIM, UBA, or UBC domains present within many components of the internalization and lysosomal targeting machinery. This complex array of signals and recognition proteins ensures the dynamic but accurate distribution of transmembrane proteins to different compartments of the endosomal-lysosomal system. PMID- 12651741 TI - Temporal and spatial regulation in prokaryotic cell cycle progression and development. AB - Bacteria exhibit a high degree of intracellular organization, both in the timing of essential processes and in the placement of the chromosome, the division site, and individual structural and regulatory proteins. We examine the temporal and spatial regulation of the Caulobacter cell cycle, bacterial chromosome segregation and cytokinesis, and Bacillus subtilis sporulation. Mechanisms that control timing of cell cycle and developmental events include transcriptional cascades, regulated phosphorylation and proteolysis of signal transduction proteins, transient genetic asymmetry, and intercellular communication. Surprisingly, many signal transduction proteins are dynamically localized to specific subcellular addresses during the cell division cycle and sporulation, and proper localization is essential for their function. The Min proteins that govern division site selection in Escherichia coli may be the first example of a system that generates positional information de novo. PMID- 12651742 TI - Accident prevention. PMID- 12651743 TI - Cloacal exstrophy. PMID- 12651744 TI - "Stay off the greens". PMID- 12651745 TI - Semeiology--a well established and challenging paediatric speciality. AB - "This week's mystery comes from seven-year-old Callum from Middlesex who, when walking a variable distance, develops an ache around the knees and down his calf of such severity that he has to stop, when it disappears immediately. He also used to wake at night complaining of 'pains in the leg' when younger. This has now improved. This is a most puzzling case, as the relationship to exercise is strongly suggestive of interruption of the blood supply or claudication, though this would not explain the nocturnal pain. Callum has been investigated from top to toe by paediatric specialists, who have been unable to reach a diagnosis. So he, and his mum, will be more than grateful if anyone out there has any suggestions." (Dr James Le Fanu, Doctor's Diary, Daily Telegraph, 2 April 2002) PMID- 12651746 TI - Growth hormone therapy in the Prader-Willi syndrome. PMID- 12651747 TI - The term diplegia should be abandoned. PMID- 12651748 TI - Evaluation of cultural competence and antiracism training in child health services. AB - AIMS: To evaluate the acceptability and effectiveness of cultural competence and antiracism training to professionals providing services to ill or disabled children. METHODS: Immediate post-training and retrospective questionnaire survey of trainees. Main outcome measures were acceptability; perceived relevance to practice; previous training in this area; perceived impact on professionals' confidence in providing care to diverse communities; and reported changes in behaviour and practice. RESULTS: Cultural competence and antiracism training has been neglected in the health sector but is well received by professionals. It is a positive experience for trainees and perceived to be relevant to their practice. Appropriate and non-threatening training in cultural competence changes attitudes, behaviours, and practice, including promoting good practice in communication across linguistic and cultural differences. CONCLUSIONS: Appropriate cultural competence and antiracism training is both effective and acceptable in child health services. PMID- 12651749 TI - General Certificate of Secondary Education performance in very low birthweight infants. AB - AIM: To compare children of very low birth weight with matched controls for their performance in the General Certificate of Secondary Education (GCSE). METHODS: GCSE examination results of 167 children of birth weight < or =1500 g attending mainstream schools and without clinical disability and 167 individually matched classroom controls were analysed. RESULTS: In 143 instances, both children of a matched pair were entered for examination in one or more GCSE subjects. The total points score obtained was greater in the comparison group than in the index cases (difference between means 4.45: 95% CI 0.95 to 7.94; p = 0.01). The mean point score per examination subject was also significantly greater in the comparison group than in the index cases (mean of differences 0.43: 95% CI 0.12 to 0.73; p < 0.01). CONCLUSIONS: As the children were closely matched for school and several social variables, factors acting during fetal or early postnatal development of very low birthweight infants probably compromise performance in the GCSE examination to a greater extent than school or childhood social environmental factors. PMID- 12651751 TI - Does providing social services with information and advice on immunisation status of "looked after children" improve uptake? AB - We prospectively assessed whether providing social services with information on the immunisation status for a cohort of looked after children in the care of an urban unitary authority in England improved uptake rates. The provision of such information did not improve immunisation coverage in these children. PMID- 12651753 TI - Early detection of hypertension in general practice. AB - We suggest a formula to aid diagnosis of hypertension in children and adolescents in general practice. The values obtained from this formula were found to correlate well with the values in the charts recommended by the Update of 1987 Task Force report on high blood pressure in children and adolescents. PMID- 12651755 TI - Thirsty Thursday. PMID- 12651754 TI - National guideline for the management of suspected sexually transmitted infections in children and young people. AB - The Children Act 1989 defines a child as "a person who has not yet reached 18 years of age." In England, Wales, and Scotland the present age of consent for heterosexual and homosexual sex is 16 years and in Northern Ireland it is 17 years. The proportion of young people who report heterosexual intercourse before the age of 16 years increased in the 1990s compared with the previous decade. PMID- 12651757 TI - Comparative efficacy of terbutaline sulphate delivered by Turbuhaler dry powder inhaler or pressurised metered dose inhaler with Nebuhaler spacer in children during an acute asthmatic episode. AB - AIMS: To compare the efficacy of terbutaline sulphate delivered via Turbuhaler with a pressurised metered dose inhaler (pMDI) connected to Nebuhaler spacer in a population of asthmatic children presenting to emergency departments because of an acute episode of asthma. METHODS: Randomised double blind, double dummy, parallel study of acute asthma in the emergency department. A total of 112 children (6-16 years), who had a diagnosis of asthma, a baseline FEV1 of 25-60% of predicted normal value (PNV), and the ability to perform spirometry were studied. Patients received two doses of 0.5 mg/10 kg (maximum 2.0 mg) of terbutaline sulphate at time 0 minutes and time 30 minutes. The two groups were also stratified into subgroups based on FEV1: 25-45% and 45.1-60% PNV. FEV1 before treatment and at two 15-minute intervals after each treatment was the main outcome measure. PIF, PEF, heart rate, SpO2, and tremor were also measured at these times. RESULTS: Both the Turbuhaler and pMDI+Nebuhaler groups showed significant increases from baseline to final value in their FEV1 results, 49% and 50% change from baseline to t = 60 min, respectively (p < 0.001) using last value carried forward. No significant difference was found between the two groups for these results. Subanalysis of the stratified groups revealed similar results. In addition, no significant difference was found in the group and subgroup comparisons for heart rate, SpO2, and tremor. CONCLUSION: Results show that Turbuhaler and pMDI+Nebuhaler are similar in terms of benefit and side effects in the treatment of acute moderate to severe asthma attacks in this study population. PMID- 12651758 TI - Vulvovaginitis in prepubertal girls. AB - This retrospective study evaluated the clinical features and findings in bacterial cultures and in microscopic examination of vaginal secretions in 80 prepubertal girls, aged 2-12 years, with vulvovaginitis. Vaginal secretions were obtained directly from the vagina with a sterile catheter carefully inserted into the vagina. Pathogenic bacteria were isolated in 36% of cases. In 59% of these cases the isolated pathogen was group A beta-haemolytic streptococcus. Candida was not found in any of the patients. The finding of leucocytes in vaginal secretions as an indicator for growth of pathogenic bacteria had a sensitivity of 83% and a specificity of 59%. Antimicrobial treatment should therefore be based on bacteriological findings of vaginal secretions and not on the presence of leucocytes alone. PMID- 12651760 TI - Metrorrhagia and precocious puberty revealing primary hypothyroidism in a child with Down's syndrome. AB - We report a child with Down's syndrome in whom metrorrhagia and precocious puberty revealed primary autoimmune hypothyroidism. The patient had a decreased growth velocity, exaggerated weight gain, bone age delay, and bilaterally enlarged multicystic ovaries. Delays in the diagnosis and treatment of hypothyroidism can lead to this peculiar presentation. PMID- 12651759 TI - Are tablets a practical source of protein substitute in phenylketonuria? AB - BACKGROUND: A phenylalanine-free amino acid based protein substitute is necessary to provide the major source of protein in phenylketonuria (PKU). Protein substitutes in PKU are usually given as drinks. These are unpalatable and compliance is often poor. Tablets containing a suitable mixture of phenylalanine free amino acids (Aminogran Food Supplement, UCB) are now available. AIMS: To compare the effectiveness and acceptability of these tablets with conventional protein substitute drinks. METHODS: Twenty one subjects with PKU, aged 8-25 years, participated in a randomised crossover study. During one phase, subjects received at least 40% of their protein substitute requirements from the amino acid tablets and the rest from their usual protein substitute tablets. During the other phase, they received their usual protein substitute. Each period lasted 12 weeks. Blood phenylalanine concentrations were measured at least once every two weeks and other plasma amino acids were measured at the beginning, at crossover, and at the end of the study. The subjects kept a diary of all protein substitute taken. RESULTS: Compliance appeared to be better with the new tablets than with patients' usual protein substitutes. Ninety per cent (18/20) recorded that they took the tablets as prescribed, compared with 65% (13/20) fully compliant with their usual protein substitute. Moreover, plasma phenyalanine was lower on the amino acid tablets, and the median difference in blood concentrations between the two groups was 46 micro mol/l (95% CI 14.8 to 89.0, p = 0.02). Tyrosine increased by a median of 16 micro mol/l daily on the amino acid tablets (95% CI 7.1 to 40.5, p = 0.01). Most subjects (70%) preferred incorporating the new tablets into their usual protein substitute regimen. CONCLUSIONS: Amino acid tablets are an effective and relatively popular protein substitute in older children, teenagers, and adults with PKU. PMID- 12651761 TI - Hyperinsulinism and overgrowth without obesity. AB - We report a 5 year old girl with postnatal overgrowth (height velocity >97th centile), hyperinsulinaemia, and increased insulin-like growth factor 1 for age, without evidence of bioactive or immunoreactive growth hormone excess or pituitary abnormality. Although her overgrowth may be a result of hyperinsulinism, her serum contains a factor (neither insulin nor IGF-1) which is able to stimulate the proliferation of lymphocyte precursors, and this could also account for the overgrowth. Over the course of two years observation she has developed acanthosis nigricans and diabetes mellitus. PMID- 12651763 TI - Psychological management of two cases of self injury on the paediatric intensive care unit. AB - Self injury has not been previously reported in an intensive care setting. Two cases are presented of ventilator dependent children with high spinal cord lesions who exhibited an unusual form of self mutilation, namely lip biting. The key to extinguishing this behaviour was to address the children's psychological needs. PMID- 12651764 TI - Anti B cell targeted immunotherapy for treatment of refractory autoimmune haemolytic anaemia in a young infant. AB - We report the case of an 8 week old infant with fulminant autoimmune haemolytic anaemia refractory to conventional immunomodulating treatment. Massive haemolysis resulted in cardiac decompensation and acute renal failure which necessitated mechanical ventilation and peritoneal dialysis. Rituximab, a chimeric anti-CD20 monoclonal antibody, halted progression of the haemolytic process, but the patient died of acute viral pneumonia and disseminated fungal infection. Earlier introduction of rituximab might have prevented the renal complications. Paediatricians should be aware of this useful therapeutic tool for treatment of refractory autoimmune haemolytic anaemia and balance its use against the risk of potential life threatening infection. PMID- 12651765 TI - Anhidrotic ectodermal dysplasia and immunodeficiency: the role of NEMO. AB - Anhidrotic (hypohidrotic) ectodermal dysplasia associated with immunodeficiency (EDA-ID; OMIM 300291) is a newly recognised primary immunodeficiency caused by mutations in NEMO, the gene encoding nuclear factor kappaB (NF-kappaB) essential modulator, NEMO, or inhibitor of kappaB kinase (IKK-gamma). This protein is essential for activation of the transcription factor NF-kappaB, which plays an important role in human development, skin homoeostasis, and immunity. PMID- 12651766 TI - Corticosteroid treatment of severe, non-responsive Clostridium difficile induced colitis. AB - Clostridium difficile can cause inflammatory diarrhoea and colitis by disrupting normal colonic flora. Corticosteroids are effective against diarrhoea associated with inflammatory bowel disease, but their effectiveness in treating inflammatory diarrhoea of C difficile has not been reported. In this preliminary report, we describe the use of corticosteroids in a child with severe C difficile diarrhoea and colitis refractory to standard treatments. PMID- 12651767 TI - Hepatosplenic cat scratch disease treated with corticosteroids. AB - A 3 year old girl with hepatosplenic cat scratch disease developed prolonged fever unresponsive to treatment with multiple antibiotics, including gentamicin, azithromycin, rifampin, and ciprofloxacin. Fever resolved with corticosteroid treatment. PMID- 12651768 TI - Bladder dysfunction in Duchenne muscular dystrophy. AB - Although bladder function is thought to be unaffected in Duchenne muscular dystrophy, 46/88 boys interviewed had urinary problems. Nine underwent video urodynamics, showing in eight a small capacity, hyperreflexic bladder, and in the ninth (post spinal surgery) hyperreflexia and detrusor sphincter dyssynergia. Urinary dysfunction is a treatable feature of DMD. PMID- 12651769 TI - An unusual life threatening cause of torticollis in a child. PMID- 12651770 TI - Syncope in childhood. AB - Syncope in childhood is very common. The vast majority of episodes are benign, and are due to neurocardiogenic syncope. Only a minority are due to something potentially more serious or life threatening. The diagnosis and differentiation of benign from more serious causes of syncope is made primarily by the history. Investigations are often unfruitful. The mainstay of management in neurocardiogenic syncope is reassurance. An increase in dietary fluid and salt can be helpful. Drug treatment is reserved for those with more frequent and severe attacks. Cardiac pacemakers should be reserved for those with very severe symptoms who are refractory to drug therapy. PMID- 12651771 TI - Update on childhood rhabdomyosarcoma. AB - The overall survival of childhood rhabdomyosarcoma has improved dramatically over the past 10 years. Early diagnosis and appropriate referral to a specialised centre leading to an accurate and timely diagnosis reflects on overall outcome. Recent molecular studies have identified different biological subtypes resulting in the recognition of poorer subgroups and allowing more appropriate treatment to be administered. Clinical trials remain the cornerstone to further improve outcome, now carried out on an international basis. PMID- 12651772 TI - Safety and success of exhaled breath condensate collection in asthma. AB - BACKGROUND: Exhaled breath condensate (EBC) is a rapidly expanding area of research to study airway inflammation through the detection of volatile and non volatile substances in the airways. AIMS: To determine the safety and feasibility of EBC procedure in a group of children with asthma of varying severity. METHODS: In a cross sectional study of children aged 4-17 years, 18 healthy and 91 asthmatic children (69 in stable condition and 22 with asthma exacerbation) underwent the EBC procedure. Outcomes assessed included completion of the procedure, decrease in FEV1, change in fractional exhaled nitric oxide (FE(NO)), and adverse effects. No pretreatment with beta2 agonists was given. All children were able to successfully complete the EBC procedure. RESULTS: Median fall in FEV1 after the procedure was -1% (IQR -3.5, 1.8) in asthmatics and was comparable to that observed in healthy children. In only one asthmatic child did the drop in FEV1 exceed 12%. No significant changes in FE(NO) were observed after EBC. CONCLUSION: This study suggests that EBC is a simple and well tolerated method for evaluating biological samples from the lower airway. The procedure was safe in children with asthma exacerbation, and the success rate was 100% in children aged 4 years and above. PMID- 12651774 TI - Response to Duke et Al. PMID- 12651773 TI - The effects of obesity on pulmonary function. AB - AIMS: (1) To determine the predominant pulmonary function abnormality in our population of obese children; and (2) to assess the correlation between the severity of lung function impairment and the degree of obesity as assessed by dual energy x ray absorptiometry (DEXA). METHODS: Sixty four obese patients underwent physical examination, standardised pulmonary function tests (spirometry, lung volumes, and single breath diffusion capacity for carbon monoxide), and DEXA scan measurements. The trunk and subtotal (total - head) body fat mass were used as surrogate index of body adiposity. RESULTS: Sixteen girls and 48 boys with median age and body mass index (BMI) of 12 years (interquartile range (IQR): 10-14) and 30.1 kg/m2 (IQR: 27.2-32.8) respectively were studied. None of the patients had clinical evidence of cardiopulmonary disease. Reduction in functional residual capacity (median FRC 93% predicted, IQR: 68.5-116.5%) and impairment of diffusion capacity (median DLco 83.5% predicted, IQR: 70.0-100.7%) were the most common abnormalities in our cohort, being observed in 30 (46%) and 21 (33%) patients respectively. Obstructive ventilatory impairment was found in three patients. There was significant negative correlation between the degree of reduction of FRC but not DLco with DEXA scan measurements, but such a relation was not found when BMI was used as the indicator of obesity. CONCLUSION: Reduction in FRC and diffusion impairment were the commonest abnormalities found in our cohort of obese patients. Reduction in static lung volume was correlated with the degree of obesity. PMID- 12651775 TI - Hypoxaemia in children: "abnormal" values may be misleading. PMID- 12651776 TI - Refugee children: don't replace one form of severe adversity with another. PMID- 12651778 TI - Ketoacid levels may alter osmotonicity in diabetic ketoacidosis and precipitate cerebral edema. PMID- 12651779 TI - Pharmacogenomic can give children safer medicines. PMID- 12651780 TI - What is the predictive value of urinary symptoms for diagnosing urinary tract infection in women? AB - OBJECTIVES: Our aim was to determine the probability of correctly diagnosing urinary tract infections (UTIs) from urinary symptoms and signs, studying their sensitivity, specificity and likelihood ratio (LR) when clinical history, signs and reactive strip test results are taken into account. METHODS: An epidemiological analysis with a diagnostic and clinical orientation was carried out in a primary health care setting. The subjects comprised 343 women > or =14 years of age who consulted their family physician for incident urinary tract symptoms. A guided medical examination was carried out using a check-list formulary, reactive strip test, urine culture and the clinical course of all patients. RESULTS: The pre-test probability of having UTI among patients with incident urinary symptoms is 0.484 [95% confidence interval (CI) 0.431-0.536]. Positive LRs for UTI are: painful voiding 1.31 (95% CI 1.12-1.54), urgency 1.29 (95% CI 1.12-1.50), urinary frequency 1.16 (95% CI 1.06-1.28) and tenesmus 1.16 (95% CI 1.02-1.32). The probability of UTI is reduced by the presence of genital discomfort, dyspareunia, vaginal discharge, positive lumbar fist percussion and perineal discomfort. Nitrites on the urine reactive strip test increase the probability of UTI by >5 times, moderate pyuria increases it by >1.5 times, and the presence of both findings does so by >7 times. CONCLUSIONS: In women with urinary symptoms, a thorough clinical examination, together with performance of a reactive strip test during the office visit, improves the chances of detecting UTI. PMID- 12651781 TI - Rapid test, throat culture and clinical assessment in the diagnosis of tonsillitis. AB - OBJECTIVES: Our aim was to identify a group of patients with clinical signs of tonsillitis to whom antibiotics could be prescribed without further diagnostic actions, and to compare the outcome of clinical assessment with the result of an antigen detection test using culture as the gold standard. METHODS: During two winter months, patients aged > or =4 years attending for sore throat at three primary health care centres in Malmo, Sweden, were examined. Odds ratios, sensitivities, specificities and predictive values were calculated for clinical assessment and for an antigen detection test. RESULTS: Among the 169 participating patients, growth of group A beta-haemolytic streptococci (GAS) was found in 53 cultures, and 23 patients (14%) were clinically assessed as 'absolutely positive', representing positive clinical assessment. Nineteen had positive cultures for GAS. The sensitivity, specificity and predictive positive and negative value for the antigen detection test were 82, 96, 90 and 93%, respectively, and for positive clinical assessment 36, 97, 83 and 77%. CONCLUSION: It is possible to identify a small group of patients with convincing signs of tonsillitis in which the specificity as well as the predictive positive value of the rapid test and the clinical assessment are close to equal. Antibiotics may be prescribed to these patients without further diagnostic actions. PMID- 12651782 TI - Women's knowledge of and attitudes towards hormone replacement therapy. AB - BACKGROUND: Hormone replacement therapy (HRT) could benefit women who have reached the natural menopause, have had a hysterectomy or have a family history of osteoporosis. OBJECTIVE: Our aim was to monitor changes in women's knowledge of, and attitudes towards, HRT since 1991. METHODS: The study was a repeat of a postal survey conducted in 1991 in the Grampian region in the North East of Scotland. Six hundred women, aged 20-69 years, were selected randomly from the eight Local Health Care Co-operatives in Grampian, Scotland. The main outcome measures were women's knowledge of HRT, their attitudes towards it and the percentage of users, past users and never users within the sample. RESULTS: A 79% response rate was achieved. Overall, 17% of post-menopausal women were current takers (increased from 9% in 1991), 22% were previous takers (increased from 7%) and 61% were never takers (decreased from 84%). This increase in ever use of HRT was more pronounced in the less educated women (increase of 24% since 1991) compared with the more educated (increase of 13%). Almost half (48%) of post menopausal women had considered taking HRT (25% increase). However, of never users, the majority (86%) had never considered HRT and had not discussed it with a doctor. Attitudes towards the menopause remained positive, although knowledge of the effects of HRT and of risk factors for osteoporosis had decreased. Forty two per cent of never users would be persuaded to take HRT if they knew it would not cause any problems, and 52% would be persuaded to take HRT on the recommendation of a doctor. CONCLUSIONS: Since 1991, HRT use increased overall; this increase was greater in the less educated women. However, the majority of post-menopausal women remain never users, and many were unaware of HRT. Conflicting research evidence since 1991 on the risks and benefits of HRT may account for the decrease in the women's knowledge of the effects of HRT. PMID- 12651783 TI - A qualitative study of GPs' attitudes to drug misusers and drug misuse services in primary care. AB - BACKGROUND: Due to a growing drug misuse problem in Scotland, GPs increasingly are being called upon to deal with and treat misusers. With the publication of government guidelines in 1999, debate on this subject has intensified, with some GPs questioning their role in the treatment of drug misuse. OBJECTIVES: This study explored, in depth, GPs' views and experiences of providing services for drug misusers such as methadone maintenance, use of guidelines and shared care schemes. Factors influencing their treatment of drug misusers were covered. METHODS: Forty-eight semi-structured GP telephone interviews were conducted and their responses recorded, transcribed and analysed using qualitative content analysis. The sample was purposive and included a spread of individuals from varying age groups, levels of involvement with drug misuse and training, as well as gender and health board. Transcripts were validated with a 50% sample of interviewees, allowing them to correct any opinions they felt were not represented accurately. RESULTS: GPs increasingly accept a role in the treatment of drug dependency, although a shared care model was preferred due to the workload implications of this patient group. A drawback of shared care was long waiting lists. Patient behaviour emerged as a strong theme influencing treatment, and safety concerns were evident. Views of methadone maintenance were mixed, and it was considered as the only real option available. CONCLUSION: Themes identified from the interviews reinforced the patterns of past research. GPs are becoming more confident and comfortable with misusers, and more positive towards methadone and methadone maintenance treatment, but still feel that they lack the necessary knowledge and skills. PMID- 12651784 TI - The needs of patients receiving depot antipsychotic medication within primary care. AB - BACKGROUND: Depot antipsychotic medication is a vital treatment for many people with schizophrenia. Many patients receive this medication from primary care, and have little or no contact with specialist mental health services. OBJECTIVE: The aim of the present study was to compare the characteristics and needs of patients receiving depot medication within primary care with those receiving their medication from specialist mental health services. METHODS: A total of 58 patients were identified from four primary care teams. Data were collected from medical records, and staff interviews were conducted using standardized interview schedules. RESULTS: Patients receiving medication from primary care had more than twice as many health and social needs compared with those receiving medication from specialist services. CONCLUSIONS: There are advantages for patients to receive depot medication from primary care, but the needs of the patients must be reviewed regularly, and primary care staff must have easy access to specialist mental health services. PMID- 12651785 TI - A qualitative study of Australian GPs' attitudes and practices in the diagnosis and management of attention-deficit/hyperactivity disorder (ADHD). AB - BACKGROUND: The importance of general practice involvement in the care of attention-deficit/ hyperactivity disorder (ADHD) is increasing due to the rising numbers of patients who present with the disorder. It has been suggested by consensus bodies that GPs should be identifying and referring patients at the severe end of the ADHD spectrum and managing those with less severe symptoms. However, GPs' views of their role in ADHD care are unknown. OBJECTIVE: Our aim was to explore the attitudes and practices of Australian GPs towards the diagnosis and management of ADHD. METHODS: We conducted a series of focus groups to explore GPs' beliefs regarding the causes of ADHD, their perceived role in ADHD diagnosis and management and their views on the role of behaviour therapies and pharmacotherapies in ADHD management. The subjects were 28 GPs in six focus groups. RESULTS: GPs in this study did not want to be the primary providers of care for patients with ADHD. Participants indicated a preference to refer the patient to medical specialists for diagnosis and treatment of ADHD, and expressed low levels of interest in becoming highly involved in ADHD care. Concerns about overdiagnosis and misdiagnosis of the disorder, diagnostic complexity, time constraints, insufficient education and training about the disorder, and concerns regarding misuse and diversion of stimulant medications were the reasons cited for their lack of willingness. CONCLUSIONS: The Australian GPs in this study identify a role for themselves in ADHD care which is largely supportive in nature, and involves close liaison with specialist services. PMID- 12651786 TI - Referral for genetic counselling during pregnancy: limited alertness and awareness about genetic risk factors among GPs. AB - BACKGROUND: In many countries, GPs play a key role in the referral to other medical specialists. Referral for reproductive genetic counselling during a pregnancy of women with a genetic risk factor already present before pregnancy has many disadvantages. Nevertheless, some 10-20% of the counsellees who attend a Department of Clinical Genetics for the first time are pregnant. OBJECTIVES: We aimed to explore the role of the GP in referring women for genetic counselling during, instead of before a pregnancy. METHOD: The GPs of 100 pregnant women who received genetic counselling were invited to participate in the study and asked to complete a questionnaire. The topics were: initiation and discussion of aspects of referral to the Department of Clinical Genetics; reasons for the referral during, instead of before a pregnancy; knowledge of genetic counselling; attitudes towards genetic counselling before a pregnancy; and attitudes towards abortion. RESULTS: To our surprise, 29% of the GPs indicated that they had not been involved in the referral to the Department of Clinical Genetics at all. Furthermore, the referral was initiated by the patient herself in most cases (40%) and by the GPs in 31% of the cases. Of the GPs who were involved in the referral, most of them (79%) talked to their patients to different extents about what to expect from their visit to the Department of Clinical Genetics; however, potential choices after an adverse outcome at prenatal diagnosis were discussed less often (60%). The main reason for referring the patient during, instead of before her pregnancy was because the GP was unaware of a potential risk factor before pregnancy (71%) and, consequently, never had a chance to talk about a referral before (71%). Other reasons for referral during pregnancy mentioned by the GPs were reassuring the patient about the health of her unborn child (32%) and the wish of the patient to be referred during pregnancy (31%). GPs considered their knowledge of clinical genetics to be limited (mean score 5, on a scale from 0 to 10). The majority of the GPs were in favour of genetic counselling taking place before, instead of during pregnancy, and they had no great objections to abortion. CONCLUSIONS: During pregnancy, the gatekeeper function of the GP in the referral for genetic counselling is undermined. Limited alertness and awareness among GPs about genetic risk factors in their patients played a major role in this undermined function and in the less appropriate timing of referral. Neither insufficient knowledge nor barriers to acceptance explained this lack of alertness and awareness. We advocate the implementation of routine family history taking in general practice. PMID- 12651787 TI - Women's interest in GP-initiated pre-conception counselling in The Netherlands. AB - BACKGROUND: Pre-conception counselling has the potential to reduce pregnancy complications and congenital disorders. The timing of counselling, before conception, is crucial to maximize the benefit. As many couples are unaware of their risk status and the fact that the first period of pregnancy is crucial, they do not seek information before pregnancy occurs. To reach couples with timely information, it seems that a health care worker needs to take the initiative. In The Netherlands, the GP is in an ideal position to offer pre conception counselling. OBJECTIVE: The aim of this study was to determine the interest of women aged 18-40 in pre-conception counselling if this is offered to them by their own GP. METHOD: A cohort of women (n = 1206) received a personal letter from their own GP with an offer of pre-conception counselling. The women were requested to fill in a reply form, indicating if they were interested, might be interested (if they decided to become pregnant) or were not interested in an invitation for pre-conception counselling. When interested, they were asked to give an indication as to when they were planning a pregnancy. Women who were not interested were requested to give a reason. RESULTS: Almost 70% of the women returned the reply form. Up to the age of 29 years, at least 80% of the respondents were interested or might be interested should they decide to have children. Most women, especially the younger women, do not know exactly when they wish to become pregnant. Regardless of age, >70% of the respondents were interested. Only 11% of the respondents indicated specifically that they were not interested in advice. CONCLUSION: Women are interested in GP-initiated pre conception counselling. Further research is needed to assess the effect of programmed and systematic pre-conception counselling, offered by GPs, on pregnancy outcome and the health of the children. A randomized controlled trial to assess these effects currently is being conducted at the Department of General Practice in Leiden. PMID- 12651788 TI - Chronic pain and the use of conventional and alternative therapy. AB - BACKGROUND: Chronic pain is a common problem affecting about half of the general population. This has implications for the utilization of both conventional and alternative health services. OBJECTIVES: The aim of this study was to determine the use of conventional and alternative practitioners and medicines amongst individuals with chronic pain in the community. METHODS: A total of 2422 individuals from a previous population-based survey in the Grampian region of the UK, who agreed to participate in further research, were sent a postal questionnaire. The questionnaire enquired about the presence, type and severity of chronic pain, socio-demographic details, consultations with conventional and alternative practitioners, and the consumption of conventional and alternative medicines. The main outcome measures were the number and frequency of self reported consultations with GPs, hospital specialists, physical therapists and alternative therapists, and the consumption of prescription, non-prescription and alternative medicines amongst those with chronic pain. RESULTS: Of the 840 individuals reporting chronic pain, 67.2% had seen their GP, 34.0% a hospital specialist, 25.9% a physical therapist and 18.2% an alternative therapist in the preceding year. Prescription medicines had been taken by 58.4%, non-prescription medicines by 57.4% and alternative medicines by 15.7% of individuals with chronic pain. The majority (67.0%) of individuals with chronic pain who sought alternative health care did so in conjunction with conventional health care. Differences in consultations with practitioners and consumption of medicines were found by age, sex, socio-economic status, site of pain and severity of pain. CONCLUSIONS: Individuals with chronic pain consult their GP about their pain more than other practitioners and use conventional medicines more frequently than alternative medicines. Alternative health care is used most commonly in addition to conventional health care, although a small number of individuals with chronic pain use alternative care exclusively. The use of alternative health care amongst those with chronic pain is higher than previously estimated and suggests that the use of these services may be increasing amongst those with chronic pain. PMID- 12651789 TI - GP care for moderate to severe asthma in children: what do infrequently attending mothers disagree with and why? AB - OBJECTIVES: Our aim was to identify and account for areas of disagreement with GP care for moderate to severe child asthma among mothers who infrequently use this care. Identifying and understanding these areas of disagreement has the potential to improve child access to GP care. METHODS: This qualitative study in Auckland, New Zealand, used a general inductive approach to analyse 23 semi-structured, personal interviews during March-October 2001 with samples of 11 mothers of children with moderate to severe asthma, and 12 medical providers (10 in general practice and two in hospital emergency departments). Disagreement was defined by mothers' non-acceptance or disapproval of aspects of GP care they reported getting for child asthma. RESULTS: Mothers and providers described four areas in which some mothers disagree with aspects of GP care for child asthma. Contributing to infrequent attendance, the areas are the validity of the diagnosis, the level of service provision, the effectiveness of care and the level of respect from practice staff. These areas revealed three groups of mothers. GP factors contributing to disagreements among mothers were reported to be inconsistent care; information deficits on asthma and individual children; a lack of commitment to identifying the cause(s) and self-management of asthma in children; and an unmet need for asthma management plans that incorporate families' knowledge, goals and preferences. CONCLUSIONS: Disagreement, among mothers, with areas of GP care for child asthma contributes to non-attendance for this care. This paper identifies opportunities for GPs to keep disagreements to a minimum and facilitate access. PMID- 12651790 TI - Influence of family on acceptance of influenza vaccination among Japanese patients. AB - BACKGROUND: Influenza is a major cause of morbidity and mortality in Japan and worldwide, especially for people of >65 years old and those with high-risk medical conditions. Although the influenza vaccine is effective in reducing the morbidity and mortality, the vaccine coverage rate has not increased adequately in Japan, compared with western countries. OBJECTIVE: Our aim was to assess whether medical and personal characteristics are associated with receiving influenza vaccination in Japanese patients. METHODS: Out-patients of a city hospital were recruited for a case-control study between November 1998 and February 1999. Cases were 98 out-patients aged 18 years or older who received influenza vaccination. Controls were 112 non-vaccinated out-patients matched with cases for primary physician and date of clinic visit. The candidates were interviewed by telephone and asked to respond to a 26-item questionnaire. The data were analysed using multiple logistic regression models. RESULTS: The factors associated with the acceptance of influenza vaccination were: (i) recommendation by a family member and/or a close friend [odds ratio (OR) 17.74; 95% confidence interval (CI) 1.95-161.77]; (ii) belief in influenza vaccine efficacy (OR 10.55; 95% CI 3.42-32.49); (iii) having a family member and/or friends who had been vaccinated before (OR 6.44; 95% CI 2.37-17.50); (iv) physician's recommendation (OR 4.03; 95% CI 1.42-11.37); and (v) knowledge about the influenza vaccine (OR 3.06; 95% CI 1.02-9.20). Fear of adverse reactions (OR 0.21; 95% CI 0.07-0.66) was the sole factor associated with non-acceptance of influenza vaccine. CONCLUSION: Patients in Japan are likely to be greatly influenced by their family members or close friends in their decision of whether to accept influenza vaccination, unlike US patients who make health care decisions on their own. When implementing an influenza vaccination programme, this effect of cultural background observed in Japan should be taken into account in other countries. PMID- 12651791 TI - GPs working in solo practice: obstacles and motivations for working in a group? A qualitative study. AB - OBJECTIVE: Our aim was to analyse the obstacles and eventual motivations of solo GPs for working in group practice. METHODS: A qualitative study using 12 focus groups was carried out in primary care in French-speaking Belgium. The subjects comprised four samples of GPs: 20 GP trainers, 18 GP trainees, 25 women GPs and 25 other GPs. The focus groups were taped and transcribed. Two independent researchers carried out the analysis using the QSR NUD.IST software. RESULTS: The participants (88 GPs) did not share a common definition of group practice-in particular multidisciplinary working-the need for a common pool of patients and shared premises. Their main sources of motivation for eventually setting up a group practice were better quality of life, continuity of care and sharing professional knowledge. The main obstacles were a required agreement between colleagues, the loss of a personal patient-GP relationship, budgetary constraints, and divergent views on group practice and GPs' profession (especially true for the association of GPs from different age groups). CONCLUSION: The current study shows that GPs working solo have divergent views of group practice. However, they clearly perceive advantages to this type of association (e.g. better quality of life and continuity of care). This study also confirms the high level of stress and tiredness felt by GPs and especially senior practitioners. PMID- 12651792 TI - Quality of recording of data from patients with type 2 diabetes is not a valid indicator of quality of care. A cross-sectional study. AB - BACKGROUND: The quality of recording of clinical data in diabetes care in general practices is very variable. It has been suggested that better recording leads to improved glycaemic control. OBJECTIVES: The purpose of this study was to assess the completeness of recording by GPs of data from type 2 diabetes patients; to compare recorded and missing data; and to investigate the association between completeness and glycaemic control. METHODS: A cross-sectional survey was carried out in 52 general practices. Medical records were scrutinized for the presence of 11 variables. Examining patients through an active approach completed incomplete records. We compared recorded and unrecorded items. Completeness of recording was determined at both patient and practice levels. RESULTS: Fifty-two general practices with 1641 type 2 diabetes patients cared for by the GP participated. The frequency of absence of any particular item ranged from 20 to 70%. Weight, systolic blood pressure and HbA(1c) were slightly lower in patients with those items missing on their files, and more such patients were non-smokers (P < 0.05). The percentage of patients with unrecorded variables that exceeded target values ranged from 39 to 75. Neither at practice level nor at patient level was any association between the completeness of the data recording and HbA(1c) found. CONCLUSION: Records often were incomplete, which hampers a systematic approach to care of diabetic patients. However, the lack of association between completeness of data recording and control of glycaemia indicates that improved recording is not a valid indicator of good quality of care. PMID- 12651793 TI - Non-attendance in general practice: a systematic review and its implications for access to primary health care. AB - BACKGROUND: Non-attendance in general practice has received increasing attention over the past few years. Its relationship with access to health care has been recognized and is of particular relevance in light of the access targets set out in the NHS Plan. METHODS: The literature was searched for articles relating to non-attendance. Titles and abstracts were examined, and relevant articles obtained. Bibliographies were examined for further references. Articles that described interventions for reducing non-attendance that were comparative studies and that examined general appointments, as opposed to appointments for screening purposes for example, were of particular interest. RESULTS AND CONCLUSIONS: The epidemiology of non-attendance has been well described, but there is little work on the reasons for non-attendance. Evidence for effective interventions to improve attendance in primary care is lacking, and this may prove to be an area of research interest in the future. As well as specific interventions to reduce non-attendance, new approaches to health care access are required in order to tackle this issue. PMID- 12651794 TI - Working with Bangladeshi patients in Britain: perspectives from Primary Health Care. AB - BACKGROUND: The difficulties of ethnic minority communities in accessing appropriate primary care are well documented, but little is known about the experiences of Primary Health Care Teams (PHCTs) serving these communities, or their strategies to help patients overcome these difficulties. OBJECTIVE: The purpose of the study was to explore the PHCT perspective of working with Bangladeshi patients. METHODS: Qualitative group discussions with PHCTs were set up by four health centres in the Grangetown area of Cardiff, where a large proportion of the Bangladeshi community lives. Experiences of and attitudes to working with Bangladeshi patients were explored. Discussions were taped and transcribed for independent analysis by two researchers. Comparisons within and between PHCTs were made. RESULTS: PHCTs largely entered into full and frank discussions. Health visitors had made significantly more effort than others to get to know their Bangladeshi patients. This had costs in terms of time and effort, with no reduction in caseload. Cutting across this difference were common themes such as communication and cultural differences, and patients' difficulties in using NHS services appropriately, which caused disruption and frustration. While there was an awareness of the reasons for these difficulties, PHCTs generally were not able to allow for them because of the inflexibility of their workload and systems of working. CONCLUSIONS: Group discussions are a useful way to encourage PHCTs to reflect on their practice and share experiences. PHCTs are aware of their patients' needs and keen to explore racial awareness training and new ways of looking at how they work. However, the grind of heavy workloads makes this process unlikely without outside facilitation. PMID- 12651795 TI - Continuing professional development (CPD): GPs' perceptions of post-graduate education-approved (PGEA) meetings and personal professional development plans (PDPs). AB - BACKGROUND: Conventional post-graduate meetings-typically 'lunchtime' meetings outside practices-have been heavily criticized. Revalidation is also impending, and there has been associated pressure for the widespread introduction of personal development plans (PDPs). However, there is very little empirical evidence about the usefulness to GPs of different kinds of educational meeting or of PDPs. OBJECTIVES: Our aim was to assess the utility to GPs of different types of post-graduate meeting and PDPs. METHODS: A postal questionnaire was sent to 921 GP principals in three health authorities, who were asked to recall their most recent post-graduate education-approved (PGEA) meetings (practice-based and 'outside') and the latest major learning 'undertaken' in their PDP. RESULTS: A total of 698 GPs (76%) returned questionnaires. A substantial minority (208; 30%) had a PDP. Most had undertaken education recently [median time elapsed (weeks): meeting 'outside' practice, 4; 'practice-based', 5; PDP, 3]. Education had not changed clinical practice for many GPs ('practice-based' 39% reported no change; 'outside' meetings 50% and PDPs 57%). A change in practice after a practice meeting was related to relevance to everyday practice [disagree/neutral, agree, strongly agree odds ratios: 1.00, 4.22 (95% CI 2.1-8.6) and 5.9 (2.6-13.3), respectively], to lecturer factors (enthusiasm, summarizing important points, handouts) and to social enjoyment. PDPs were less likely to be perceived relevant to practice (practice-based meeting, 'outside' meeting, PDPs: 89, 87 and 72%, respectively), as a break from practice (54,72 and 18%), good socially (63, 72 and 15%), good for professional networking (54, 70 and 19%) and glad to have done it (84, 86 and 44%). Being glad to use a PDP was more likely if the learning was clinically relevant, a break from practice, and incorporated professional networking. CONCLUSION: Changes in practice after post-graduate meetings are not only related to clinical relevance and lecturer factors, but also to professional and social factors. PDPs may not be providing better learning opportunities or enjoyment than traditional meetings, although GPs who are glad to use PDPs incorporate clinical relevance, a break from practice and networking. Post graduate tutors should probably continue to support and monitor the lecturer quality and clinical relevance of a balanced portfolio of both practice-based and 'outside' meetings. PMID- 12651796 TI - Do clinical practice education groups result in sustained change in GP prescribing? AB - BACKGROUND: Concern has been expressed at the poor uptake of evidence into clinical practice. This is despite the fact that continuing education is an embedded feature of quality assurance in general practice. There are a variety of clinical practice education methods available for dissemination of new evidence. Recent systematic reviews indicate that the effectiveness of these different strategies is extremely variable. OBJECTIVE: Our aim was to determine whether a peer-led small group education pilot programme used to promote rational GP prescribing is an effective tool in changing practice when added to prescribing audit and feedback, academic detailing and educational bulletins, and to determine whether any effect seen decays over time. METHODS: A retrospective analysis of a controlled trial of a small group education strategy with 24 month follow-up was carried out. The setting was an independent GPs association (IPA) of 230 GPs in the Christchurch New Zealand urban area. All intervention and control group GPs were already receiving prescribing audit and feedback, academic detailing and educational bulletins. The intervention group were the first 52 GPs to respond to an invitation to pilot the project. Two control groups were used, one group who joined the pilot later and a second group which included all other GPs in the IPA. The main outcome measures were targeted prescribing data for 12 months before and 24 months after each of four education sessions. RESULTS: An effect in the expected direction was seen in six of the eight key messages studied. This effect was statistically significant for five of the eight messages studied. The effect size varied between 7 and 40%. Where a positive effect was seen, the effect decayed with time but persisted to a significant level for 6-24 months of observation. CONCLUSION: The results support a positive effect of the education strategy on prescribing behaviour in the intervention group for most outcomes measured. The effect seen is statistically significant, sustained and is in addition to any effect of the other pharmaceutical educational initiatives already undertaken by the IPA. PMID- 12651797 TI - Effect of extension of postgraduate training in general practice on the acquisition of knowledge of trainees. AB - BACKGROUND: Postgraduate training in general practice aims to develop clinical competence. However, little is known about its effect on trainees' development of competence. OBJECTIVE: Our aim was to investigate whether 3 years of postgraduate training in general practice leads to a higher level of knowledge than 2 years training while maintaining the same structure and educational objectives. METHODS: This retrospective study had a mixed longitudinal design. Trainees of the Dutch postgraduate training in general practice participated. Knowledge was assessed by written progress testing of knowledge relevant to general practice embedded in real life situations. Test results were collected from 1992 to 1999. The results of trainees who received the 2-year and 3-year curriculum were compared. RESULTS: Both curricula started with the same entrance level and showed the largest acquisition of knowledge at the start and towards the end of training. Both curricula showed stagnation in growth at the end of the training period in which trainees rotate through hospitals, nursing homes and mental health institutions. The level of knowledge at the end of training was significantly higher for the 3-year curriculum. CONCLUSION: This study shows that postgraduate training contributes to an increase in knowledge and that a 3-year programme leads to a higher level of knowledge than a 2-year programme. The stagnation in growth found at the end of rotations through hospitals, nursing homes and mental health institutions questions the impact of these rotations on the development of competence and the efficacy of the training as a whole. Further study is needed to draw more firm conclusions. PMID- 12651798 TI - Medical student interviewing: a randomized trial of patient-centredness and clinical competence. AB - BACKGROUND: It is widely accepted that the quality of doctor interviewing skills is an important determinant of health care outcomes. Two interviewing skills contributing significantly to optimal health outcomes are the clinical competence of the interviewer and the use of patient-centred techniques. However, the relative importance of these to health outcomes is unknown. OBJECTIVE: The purpose of this study was to examine the relative effect on maternal recall and satisfaction of medical student clinical competence and use of patient-centred interview techniques. METHODS: Sixty-nine mothers of children attending the Paediatric Medical Out-patient Clinic, Women's and Children's Hospital, South Australia agreed to participate in the study, with 60 successfully completing the study. They viewed two of four standardized 'medical student' interview videotapes in which the level of clinical competence and patient-centredness were varied independently. All other interview variables were controlled. Each mother rated the interviews by questionnaire (balanced incomplete block design, each interview rated by 30 mothers). Maternal satisfaction with the 'student' interview was measured using the Medical Interview Satisfaction Scale (MISS) and the Interpersonal Skills Rating Scale (IPS). Maternal recall of interview information was assessed by questionnaire, with maternal answers coded independently before analysis. RESULTS: Significant, independent effects of clinical competence and patient-centredness were observed with both MISS (F = 42.1, P < 0.0001) and IPS (F = 49.3, P < 0.0001) scores. The effect of clinical competence was stronger than that of patient-centredness. Maternal recall for specific information was greater for some items following the more clinically competent interviews, but was lower for others. There was no association between maternal recall and the level of patient-centredness demonstrated by the 'student'. CONCLUSION: Clinical competence was a more significant determinant of maternal evaluations of medical student interviews and maternal recall of information than was patient-centredness. High levels of both patient-centredness and clinical competence were associated with the highest maternal satisfaction. PMID- 12651800 TI - Informed consent for screening. PMID- 12651801 TI - Mass spectrometry-based diagnostics: the upcoming revolution in disease detection. PMID- 12651802 TI - Standardization as a private enterprise. PMID- 12651803 TI - The search for a biomarker of cardiac ischemia. PMID- 12651804 TI - Surveillance of insulin resistance in children. PMID- 12651805 TI - Comparison of two CYP2D6 genotyping methods and assessment of genotype-phenotype relationships. AB - BACKGROUND: There have been no published reports comparing the CYP450 GeneChip microarray assay with more standard methods of genetic testing. METHODS: We collected 20-mL blood samples from 236 volunteers for DNA isolation and testing before each individual ingested 60 mg of dextromethorphan, and collected their urine. CYP2D6 alleles *3 to *7, *9, *17, and *41, and multiple CYP2D6 gene copies were tested by allele-specific PCR (AS-PCR), whereas alleles *2 to *4 and *6 to *11 were tested by the Affymetrix CYP450 GeneChip assay. Five of the CYP2D6 alleles (*3, *4, *6, *7, and *9) were tested by both AS-PCR and the CYP450 GeneChip assay in an independent and blinded fashion in 232 of the 236 healthy volunteers. The combined CYP2D6 genotype from both methods was used to divide the population into four subgroups, poor metabolizers (PMs), intermediate metabolizers (IMs), extensive metabolizers (EMs), and ultrarapid metabolizers (UMs), based on their relative function and ability to express the CYP2D6 gene. The urinary elimination of dextromethorphan was assessed in each of these CYP2D6 subgroups. RESULTS: The CYP2D6*3, *4, *6, *7, and *9 alleles showed a high degree of concordance between the CYP450 GeneChip and AS-PCR methods (>99% concordance). The mean (SD) of the log[dextromethorphan metabolic ratio (MR)] in the four CYP2D6 subgroups was PM = 0.49 (0.38); IM = -1.24 (0.53); EM = -2.35 (0.61); and UM = -2.43 (0.38). CONCLUSIONS: Oligonucleotide microarray technology is an efficient and reliable way to test for CYP2D6 gene variation based on five alleles compared by separate methods. The methodology is influenced by the quality and amount of DNA present. The log(dextromethorphan MR) is a highly variable index that appears to reflect the crude nature of the dextromethorphan MR as an indicator of CYP2D6 in vivo enzyme activity. PMID- 12651806 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry based detection of microsatellite instabilities in coding DNA sequences: a novel approach to identify DNA-mismatch repair-deficient cancer cells. AB - BACKGROUND: Inherited defects in the DNA mismatch repair system lead to increased loss or gain of repeat units in microsatellites, commonly referred to as microsatellite instability (MSI). MSIs in coding regions of critical genes contribute to the pathogenesis of DNA-mismatch repair-deficient cancers, particularly those associated with the hereditary nonpolyposis colorectal cancer syndrome (HNPCC). MSI typing is therefore increasingly used to guide the molecular diagnosis of HNPCC. METHODS: We used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to identify MSIs in mononucleotide repeats within the coding sequences of genes relevant to the pathogenesis of MSI+ neoplastic lesions. After a primer extension reaction of PCR products encompassing the microsatellites, the molecular masses of the extension products were determined by MALDI-TOF-MS. RESULTS: MSIs were detected by MALDI-TOF-MS in the GART, AC1, TGFBR2, MSH3, and MSH6 genes in neoplastic tissues and MSI+ colorectal cancer cell lines but not in MSI- control tissues. The analysis of peak-integral ratios in a single spectrum of the peaks representing insertions or deletions compared with the full-length microsatellites allowed relative quantification of MSIs. MALDI-TOF-MS-based genotyping results were confirmed by conventional DNA sequencing and electrophoresis. CONCLUSIONS: Because of its reliability, short run times, and low costs, this semiquantitative procedure represents an effective alternative, in particular for diagnostic high-throughput typing of MSIs in neoplastic lesions. PMID- 12651807 TI - Prognostic use of circulating plasma nucleic acid concentrations in patients with acute stroke. AB - BACKGROUND: At present there is no simple, accurate blood test that may be used to determine the severity of stroke or to predict mortality and morbidity in stroke patients presenting to emergency departments. METHODS: Patients with stroke-like symptoms who presented to an emergency department of a university hospital in Hong Kong were recruited for the study. DNA extracted from patients' plasma was analyzed for the beta-globin gene with a fluorescent-based PCR test. The primary outcome measures were in-hospital and 6-month mortality and morbidity using the post-stroke modified Rankin Score. RESULTS: Among the 88 consecutive patients recruited to the study, 70 (80%) had ischemic stroke, 11 (13%) had intracerebral hemorrhage, and 7 (8%) had transient ischemic attacks. Median plasma DNA concentrations taken within 3 h of symptom onset were higher in patients who died compared with those who survived at discharge (6205 vs 1334 kilogenome-equivalents/L; P = 0.03). Among patients with NIH Stroke Scale scores >8, median plasma DNA concentrations were higher in patients who died compared with those who survived to 6 months (2273 vs 968 kilogenome-equivalents/L; P = 0.002). Plasma DNA concentrations correlated with the volume of cerebral hematoma (r = 0.66; P = 0.03). Plasma DNA concentrations >1400 kilogenome-equivalents/L had a sensitivity of 100% and a specificity of 74.4% for predicting hospital mortality after stroke, and the area under the ROC curve was 0.89 (95% confidence interval, 0.80-0.94). The adjusted odds ratio for plasma DNA concentrations predicting 6-month mortality was 1.6 (1.1-2.4; P = 0.03) and for predicting 6 month post-Rankin Score >2 was 1.8 (1.0-3.3; P = 0.05). CONCLUSION: Plasma DNA concentrations correlate with stroke severity and may be used to predict mortality and morbidity in the emergency room. PMID- 12651808 TI - The d-dimer test for deep venous thrombosis: gold standards and bias in negative predictive value. AB - BACKGROUND: Because venous ultrasound (US) fails to fully image the calf veins, there is the potential for US gold standard studies to classify patients with calf deep venous thrombosis (DVT) in the nondiseased category, causing bias in test index calculations. A false increase in negative predictive value (NPV) is especially likely because calf DVT false-negative tests will be counted in the numerator along with the true-negative tests in NPV calculations. We verified the presence and magnitude of this bias for the d-dimer test. METHODS: We abstracted data on overall (calf and thigh) and thigh-only test sensitivity, specificity, and NPV from the six English language studies published between March 1995 and October 2001 that compared d-dimer to a gold standard (GS) capable of imaging both thigh and calf veins and that also stratified results by thigh and calf location. Thigh specificity and NPV were calculated classifying calf DVT patients as free of disease. RESULTS: The six studies included 81-214 participants and provided 26 comparisons of 16 different d-dimer assays to the GS. Thigh sensitivity was higher than overall sensitivity in 22 of 26 comparisons (range, 0.3 to 8.6); thigh specificity was lower than overall specificity in all comparisons (range, -0.7 to -7.8); and thigh NPV was higher than overall NPV in 22 of 26 comparisons and unchanged in 4 comparisons (range, 0.0-9.2). NPV was >95% in 20 of the thigh results but >95% in only 8 of the overall results. CONCLUSIONS: Different GS can produce clinically significant differences in test indices. Care must be taken in interpreting DVT studies that evaluate d-dimer as a rule-out test and that use US as a GS, because missed calf DVT can falsely increase the NPV. PMID- 12651809 TI - Cleaved protein S (PS), total PS, free PS, and activated protein C cofactor activity as risk factors for venous thromboembolism. AB - BACKGROUND: Although hereditary protein S (PS) deficiency is clearly associated with venous thromboembolism (VTE), the importance of low PS concentrations as a risk factor for VTE in other patients is still a matter of debate. To clarify this issue, we designed a case-control study to evaluate the role of different molecular forms of plasma PS. METHODS: We quantified plasma cleaved, total, and free PS and activated protein C (APC) cofactor activity in 87 VTE patients and 174 controls matched for age, sex, and hormonal treatment. Free PS was measured by ELISA or by enzyme-linked ligand sorbent assay (ELSA). Cleaved and total PS were measured by ELISA. RESULTS: In controls, the mean (SD) concentration of circulating cleaved PS was 39 (14) nmol/L, corresponding to 10% (3.5%) of total PS. Concentrations of cleaved PS and total PS were not significantly different in patients with VTE compared with controls. However, in our population, low free PS measured by ELISA or ELSA, as well as APC cofactor activity values were significantly associated with VTE with odds ratios (95% confidence intervals) of 2.9 (1.3-6.3), 2.5 (1.1-5.6), and 2.9 (1.3-6.4), respectively, in multivariate analyses. CONCLUSION: Phenotypic low PS detected by APC cofactor activity assay or by an assay specific for free PS should be considered a risk factor for VTE. PMID- 12651810 TI - Evaluation of human serum albumin cobalt binding assay for the assessment of myocardial ischemia and myocardial infarction. AB - BACKGROUND: Clinical diagnoses were correlated with results of a Co(II)-albumin binding assay in 167 patients treated at an emergency department of a health maintenance organization. METHODS: Patients were evaluated as being nonischemic or potentially ischemic through standard coronary disease indicators [creatine kinase (CK), CK-MB, cardiac troponin I, and electrocardiographic findings] and were tested by a Co(II)-albumin binding assay. Samples were tested anonymously, and the study was double-blinded. The sensitivity and specificity of this assay for the detection of ischemia were evaluated by ROC curve analysis. Known Co(II) binding sites on albumin were analyzed by N-terminal amino acid sequencing. RESULTS: The mean absorbance units (ABSU) +/- 2 SD for non-myocardial ischemic and myocardial ischemic individuals measured at 470 nm were 0.43 +/- 0.10 and 0.63 +/- 0.25, respectively (P <0.0001). The area under the ROC curve was 0.95 [95% confidence interval (CI), 0.92-0.99], and at a cutoff value of 0.50 ABSU, sensitivity and specificity were 88% (78-94%) and 94% (86-98%), respectively, suggesting a high distinction between the two groups. When we compared non-acute myocardial infarction (AMI) and AMI ischemic individuals, the area under the ROC curve was 0.66 (95% CI, 0.53-0.79) and was considered a poor discriminator between these two groups. N-Terminal amino acid sequencing data for purified albumin showed normal amino acid residues for six of seven high-ABSU (> or =0.70) individuals and one nonischemic individual tested. However, only one individual with a high ABSU (0.80) had two missing amino acid residues (DA) from the N terminal region. Clinical diagnosis for this patient did not reveal an ischemic event. CONCLUSIONS: The Co(II)-albumin binding test may serve as a useful diagnostic tool in emergency facilities for the assessment of myocardial ischemia. High and low ABSU were associated with myocardial ischemic individuals and non-myocardial ischemic individuals, respectively. However, the Co(II) albumin binding was a poor discriminator between ischemic individuals with and without MI. PMID- 12651811 TI - Catecholamine-synthesizing enzymes in carcinoid tumors and pheochromocytomas. AB - BACKGROUND: Serotonin is the principal endocrine product of carcinoid tumors, but simultaneously increased production of catecholamines has been described in these tumors. As it is not clear whether these tumors contain specific enzymes for catecholamine synthesis, we aimed to detect catecholamine-synthesizing enzymes [tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH), and phenylethanolamine-N-methyltransferase (PNMT)] in midgut carcinoid tumors and pheochromocytoma and to correlate enzyme expression to serotonin production as well as catecholamines and metabolites excreted in urine. METHODS: Paraffin embedded tumor specimens from 21 midgut carcinoid patients and 20 pheochromocytoma patients (10 sporadic and 10 MEN type IIa-related tumors) were stained for TH, DBH, and PNMT, using a three-step biotin-avidin-peroxidase method. RESULTS: TH was demonstrated in 9 (43%) of 21 carcinoids and in all (100%) of 20 pheochromocytomas, DBH in 8 (38%) carcinoids and in 15 (75%) pheochromocytomas, and PNMT in 7 (33%) carcinoids and in 13 (65%) pheochromocytomas. Increased urinary excretion of catecholamines and metabolites was observed in 10 (48%) carcinoid patients and in all pheochromocytoma patients. No clinically relevant association between enzyme expression and urinary excretion of catecholamines and metabolites was found. CONCLUSIONS: Catecholamine synthesizing enzymes are present in many carcinoid tumors. This finding possibly indicates the existence of a catecholamine-synthesizing pathway in carcinoids similar to that found in pheochromocytoma. PMID- 12651812 TI - New ELISA system for myeloid-related protein complex (MRP8/14) and its clinical significance as a sensitive marker for inflammatory responses associated with transplant rejection. AB - BACKGROUND: C-reactive protein (CRP), a useful marker for inflammatory diseases, is not always sensitive to inflammatory reaction in the liver or other tissues. The aim of this study was to develop a sensitive and specific method for detecting inflammatory responses associated with transplant rejection. METHODS: We developed a new, highly sensitive ELISA system for the measurement of serum human myeloid-related protein complex (MRP8/14), using monoclonal antibodies against MRP8/14, and applied it to specimens obtained from patients undergoing small intestine or liver transplantation. RESULTS: This assay could detect MRP8/14 concentrations as low as 2 micro g/L. Within-run CVs were 3.7-6.1% and between-day CVs were 5.6-8.7% for MRP8/14 concentrations of 117-3300 micro g/L. Mean recovery was 104% (range, 80-128%). We observed a marked increase in serum MRP8/14 postoperatively in most recipients of transplants, followed by an increase in CRP 1-7 days after the increase in the complex. The increase in serum MRP8/14 occurred simultaneously with permeation of lymphocytes into the transplanted tissues as a result of rejection of the graft tissues. CONCLUSIONS: Accurate measurement of serum MRP8/14 provides a useful clinical diagnostic method tool for detecting inflammation associated with rejection of transplanted tissues. PMID- 12651813 TI - Standardization of two immunoassays for human glandular kallikrein 2. AB - BACKGROUND: Measurement of human kallikrein 2 (hK2) has improved early detection and staging of prostate cancer. However, reported concentrations of hK2 among currently used assays have not been standardized in any way. We compared two hK2 assays and five different recombinant hK2 variants (rhK2) and suggest a common calibrator as an important step and putative reference substance in hK2 assay standardization. METHODS: We measured 146 sera by two hK2 assays, using assay specific calibrators to assess the difference between the two assays. Serial dilutions of five rhK2 preparations were measured repeatedly, with one preparation assigned as calibrator and the others as unknowns to define which variant provided the closest match between the two assays. This rhK2 variant was used to recalibrate both assays. We measured hK2 concentrations in the same 146 patients to evaluate the change in the difference. RESULTS: Use of assay-specific calibrators for comparison of the two assays yielded a Deming regression equation of: y = 0.789 (95% confidence interval, 0.674-0.922)x + 0.014 (0.004-0.025) micro g/L; R(2) = 0.667. Analysis of five rhK2 variants revealed that the enterokinase (ek)-rhK2 form provided the best match between both assays. Using the ek-rhK2 as a common calibrator, we observed a change in the slope of the regression curve to: y = 1.106 (0.872-1.340)x + 0.006 (-0.002 to 0.016) micro g/L; R(2) = 0.648, suggesting an increase in the mean estimate of agreement between the two assays. CONCLUSION: Calibration with a common calibrator substantially increased agreement between the assays. The ek-rhK2 variant provided the best performance of all tested rhK2 variants and should undergo mass spectrometry and amino acid analysis for exact mass determination and value assignment to evaluate its potential as a reference material for immunoassays for hK2. PMID- 12651814 TI - Standardization of immunoassays for measurement of high-sensitivity C-reactive protein. Phase I: evaluation of secondary reference materials. AB - BACKGROUND: Inflammation contributes to the development and progression of atherosclerosis, and C-reactive protein (CRP) can be used as a marker to assess risk for cardiovascular diseases. As variability among existing high-sensitivity CRP (hsCRP) assays can lead to misclassification of patients and hamper implementation of population-based medical decision points, standardization of hsCRP assays is needed. METHODS: We evaluated five proposed secondary reference materials, including two diluted preparations of Certified Reference Material 470 (CRM470), two preparations of a serum-based material with recombinant CRP added, and one serum-based material with isolated CRP added. Twenty-one manufacturers participated in the comparison with 28 different assays. We examined imprecision, linearity, and parallelism with these materials and with fresh serum. RESULTS: All materials had similar imprecision; CVs for the undiluted materials were 2.1 3.7%. None of the materials was linear across all assays. Each had between one and three cases of nonlinearity, with one preparation of CRM470 having the fewest cases of nonlinearity. Although none of the materials was parallel across all assays, the differences in slope from fresh serum were similar across all assays. CONCLUSIONS: All materials performed similarly with regard to imprecision, linearity, and parallelism. As one preparation of CRM470 had slightly better characteristics than the other materials and because CRM470 had been certified previously as a reference material for the acute-phase reactant range, it will be used in the next phase to standardize hsCRP assays. PMID- 12651815 TI - Quantitative urine particle analysis: integrative approach for the optimal combination of automation with UF-100 and microscopic review with KOVA cell chamber. AB - BACKGROUND: Automated systems have enabled the counting of particles in urine to be standardized. Their superiority over traditional sediment analysis has been well documented, but they have not gained wide acceptance. The reasons for this are that sediment analysis has been performed and interpreted for decades. Additionally, pathologic casts and other unknown particles still must be confirmed under the microscope. Furthermore, comparison between the methods has revealed outliers and thus decreased confidence in automation. METHODS: We used the standardized KOVA cell chamber system to count particles and compared the results with UF-100 flow cytometry as an alternative to traditional sediment analysis. RESULTS: We compared 252 randomly selected urine samples and obtained a review rate of 33%. Microscopic verification was necessary because of the presence of casts, yeast, sperm, dysmorphic erythrocytes, and some misclassified erythrocytes or leukocytes that were detected by incongruent dipstick results and abnormal scattergrams. We obtained correlation coefficients of 0.966 for erythrocytes and 0.935 for leukocytes. Criteria for an algorithm to identify samples that needed microscopic review were derived from comparisons between the number of particles from UF-100, dipstick results, cell chamber counting, and sediment analysis. CONCLUSIONS: Automated cell counting combined with microscopic counting with a standardized cell chamber system is useful. An objective algorithm for review criteria can be developed via systematic comparison of UF 100 flow cytometry and microscopy. Only urine samples that meet these criteria need to be confirmed microscopically. PMID- 12651816 TI - LOINC, a universal standard for identifying laboratory observations: a 5-year update. AB - The Logical Observation Identifier Names and Codes (LOINC) database provides a universal code system for reporting laboratory and other clinical observations. Its purpose is to identify observations in electronic messages such as Health Level Seven (HL7) observation messages, so that when hospitals, health maintenance organizations, pharmaceutical manufacturers, researchers, and public health departments receive such messages from multiple sources, they can automatically file the results in the right slots of their medical records, research, and/or public health systems. For each observation, the database includes a code (of which 25 000 are laboratory test observations), a long formal name, a "short" 30-character name, and synonyms. The database comes with a mapping program called Regenstrief LOINC Mapping Assistant (RELMA(TM)) to assist the mapping of local test codes to LOINC codes and to facilitate browsing of the LOINC results. Both LOINC and RELMA are available at no cost from http://www.regenstrief.org/loinc/. The LOINC medical database carries records for >30 000 different observations. LOINC codes are being used by large reference laboratories and federal agencies, e.g., the CDC and the Department of Veterans Affairs, and are part of the Health Insurance Portability and Accountability Act (HIPAA) attachment proposal. Internationally, they have been adopted in Switzerland, Hong Kong, Australia, and Canada, and by the German national standards organization, the Deutsches Instituts fur Normung. Laboratories should include LOINC codes in their outbound HL7 messages so that clinical and research clients can easily integrate these results into their clinical and research repositories. Laboratories should also encourage instrument vendors to deliver LOINC codes in their instrument outputs and demand LOINC codes in HL7 messages they get from reference laboratories to avoid the need to lump so many referral tests under the "send out lab" code. PMID- 12651818 TI - Distribution of fasting plasma insulin, free fatty acids, and glucose concentrations and of homeostasis model assessment of insulin resistance in a representative sample of Quebec children and adolescents. AB - BACKGROUND: Plasma fasting insulin and the homeostasis model assessment of insulin resistance (HOMA-IR) are markers of IR, which, at least in part, mediates the relation of obesity to increased cardiovascular risk. Increased free fatty acids (FFAs) may be involved in the pathogenesis of IR. Our objectives were to describe the distributions of fasting plasma insulin, glucose, and FFAs and HOMA IR in youth and to assess the relationship between FFAs and markers of IR. METHODS: Fasting plasma insulin, glucose, and FFAs were measured in a representative sample of Quebec youth comprising 2244 individuals 9, 13, and 16 years of age. RESULTS: In all age and sex groups, glucose exhibited remarkably tight distributions (median CV, 7.1%) in contrast to insulin, HOMA-IR, and FFAs (median CVs, 52%, 54% and 45%, respectively). For every percentile examined, 9 year-olds had lower insulin concentrations and HOMA-IR values than 13- and 16 year-olds. We observed strong correlations between insulin concentrations and HOMA-IR values, as well as close similarity in their rankings of individuals. The mean concentrations of glucose were higher in our population than in other Caucasian pediatric populations. No positive correlations were detected between FFAs and markers of IR. CONCLUSIONS: We report some of the first data on the distributions of fasting plasma insulin, HOMA-IR, and FFAs from a representative sample of youth. HOMA-IR does not appear more informative than fasting insulin as a marker of IR. Our findings on higher mean glucose concentrations in this population require confirmation in other representative samples of youth to assess whether the North American distribution of glucose concentrations is shifting positively. PMID- 12651817 TI - Identification of rat targets of anti-soluble liver antigen autoantibodies by serologic proteome analysis. AB - BACKGROUND: Anti-soluble liver antigen (SLA) autoantibodies are specific for autoimmune hepatitis type 1 and are the only immunologic marker found in 15-20% of hepatitis cases previously considered cryptogenic. Anti-SLA antibodies react with the 100 000g supernatant from rat liver homogenate, but the molecular targets remain controversial. METHODS: We characterized anti-SLA targets by one- and two-dimensional immunoblotting analysis. The recognized proteins were identified by peptide mass fingerprint analysis after matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. RESULTS: Three proteins of 35 kDa and pI 6.0, 50 kDa and pI between 6.0 and 6.5, and 58 kDa and pI between 6.5 and 7.0 were stained more intensely by anti-SLA positive sera than by control sera. After in-gel tryptic digestion, MALDI-TOF analysis of the generated peptides enabled the clear identification of N-hydroxyarylamine sulfotransferase, isoforms of alpha-enolase, and isoforms of catalase. CONCLUSIONS: Possible antigens for anti-SLA antibodies include a sulfotransferase, alpha-enolase(s), and catalase(s). Two-dimensional electrophoresis combined with mass spectrometry offers a versatile tool to identify molecular targets of autoantibodies and thus to improve diagnostic tools and the understanding of the immune process. PMID- 12651819 TI - Adiponectin: stability in plasma over 36 hours and within-person variation over 1 year. PMID- 12651821 TI - Detection of donor-specific DNA polymorphisms in the urine of renal transplant recipients. PMID- 12651820 TI - High stability of markers of cardiovascular risk in blood samples. PMID- 12651822 TI - Measurement of cortisol in small quantities of saliva. PMID- 12651823 TI - Tandem mass spectrometric determination of malonylcarnitine: diagnosis and neonatal screening of malonyl-CoA decarboxylase deficiency. PMID- 12651824 TI - IFCC method for lactate dehydrogenase measurement in heparin plasma is unreliable. PMID- 12651825 TI - Capillary electrophoresis method to measure p-aminohippuric acid in urine and plasma for the assessment of renal plasma flow. PMID- 12651826 TI - Population distributions of C-reactive protein in apparently healthy men and women in the United States: implication for clinical interpretation. PMID- 12651827 TI - Distributions of C-reactive protein measured by high-sensitivity assays in apparently healthy men and women from different populations in Europe. PMID- 12651828 TI - Simultaneous genotyping of seven single-nucleotide polymorphisms in the MDR1 gene by single-tube multiplex minisequencing. PMID- 12651829 TI - Measurement of beta1- and beta2-globulins improves detection of M-spikes on high resolution electrophoresis. PMID- 12651830 TI - Serial analysis of fetal DNA concentrations in maternal plasma in late pregnancy. PMID- 12651831 TI - Paradoxical changes in cystatin C and serum creatinine in patients with hypo- and hyperthyroidism. PMID- 12651832 TI - Quantification of glutamine in dried blood spots and plasma by tandem mass spectrometry for the biochemical diagnosis and monitoring of ornithine transcarbamylase deficiency. PMID- 12651833 TI - Exercise-induced myocardial ischemia is accompanied by increased serum creatine concentrations. PMID- 12651834 TI - Population distribution of high-sensitivity C-reactive protein among US men: findings from National Health and Nutrition Examination Survey 1999-2000. PMID- 12651836 TI - HLA-DRB1* intron-primed sequencing for haploid genotyping. PMID- 12651835 TI - Rapid fluorimetric method to detect total plasma malondialdehyde with mild derivatization conditions. PMID- 12651837 TI - False-positive acetaminophen results in a hyperbilirubinemic patient. AB - BACKGROUND: Acetaminophen was falsely detected in the plasma of a severely jaundiced patient, and a methodologic interference from bilirubin was suspected. METHODS: Acetaminophen was measured by an enzymatic method (GDS Diagnostics). The putative bilirubin interference was investigated in 12 hyperbilirubinemic specimens and in bilirubin linearity calibrators. The analytical method was modified to correct for background absorbance at a second wavelength. Hyperbilirubinemic specimens were fortified with acetaminophen to assess the effect of the interference on acetaminophen measurements. RESULTS: Acetaminophen was detected in 12 specimens from hyperbilirubinemic patients without a history of recent acetaminophen exposure. Dilution of hyperbilirubinemic specimens produced a nonproportional decrease in apparent acetaminophen concentrations, and no acetaminophen was detected when bilirubin was <50 mg/L. Background correction at a second wavelength failed to compensate for the interference. Although erroneous acetaminophen concentrations were detected in all specimens with high bilirubin, acetaminophen measurements in fortified specimens were accurate. CONCLUSION: The data are consistent with bilirubin interference in the enzymatic and/or chromogenic reactions involved in the acetaminophen method. PMID- 12651839 TI - Accuracy of cardiovascular risk estimation. PMID- 12651838 TI - William Prout: early 19th century physician-chemist. AB - In the early 19th century, the discoveries of new substances in the healthy and diseased body spawned a search for chemical explanations for physiologic phenomena to guide medical diagnosis and control therapy. William Prout's work on the nature and treatment of diseases of the urinary organs established his reputation as one of Britain's most distinguished physiological chemists. Prout was very skeptical of chemical remedies because of possible side effects, but he suggested iodine treatment for goiter. He emphasized that a satisfactory diet should include carbohydrates, fats, protein, and water. In 1824, he showed that the acid of the gastric juice was hydrochloric acid. Prout applied chemical methods and reasoning to physiology and was criticized for his view that the body's vital functions could be explained by chemistry. His remedy for lack of progress in animal chemistry was for physiologists to become chemists. Prout stimulated much discussion on atomic theory by his hypothesis that the atomic weights of all chemical elements are whole-number multiples of the atomic weight of hydrogen and that the chemical elements were condensed from hydrogen atoms. PMID- 12651840 TI - Detection of a novel 1905C-->T mutation within the dihydropyrimidine dehydrogenase gene and potential for misclassification with the exon 14-skipping mutation. PMID- 12651841 TI - Haptoglobin polymorphism and iron hemostasis. PMID- 12651842 TI - Negative thyrotropin assay interference associated with an IgGkappa paraprotein. PMID- 12651847 TI - Solution structure of human BCL-w: modulation of ligand binding by the C-terminal helix. AB - The structure of human BCL-w, an anti-apoptotic member of the BCL-2 family, was determined by triple-resonance NMR spectroscopy and molecular modeling. Introduction of a single amino acid substitution (P117V) significantly improved the quality of the NMR spectra obtained. The cytosolic domain of BCL-w consists of 8 alpha-helices, which adopt a fold similar to that of BCL-xL, BCL-2, and BAX proteins. Pairwise root meant square deviation values were less than 3 A for backbone atoms of structurally equivalent regions. Interestingly, the C-terminal helix alpha8 of BCL-w folds into the BH3-binding hydrophobic cleft of the protein, in a fashion similar to the C-terminal transmembrane helix of BAX. A peptide corresponding to the BH3 region of the pro-apoptotic protein, BID, could displace helix alpha8 from the BCL-w cleft, resulting in helix unfolding. Deletion of helix alpha8 increased binding affinities of BCL-w for BAK and BID BH3-peptides, indicating that this helix competes for peptide binding to the hydrophobic cleft. These results suggest that although the cytosolic domain of BCL-w exhibits an overall structure similar to that of BCL-xL and BCL-2, the unique organization of its C-terminal helix may modulate BCL-w interactions with pro-apoptotic binding partners. PMID- 12651848 TI - The membrane domain of the Na+-motive V-ATPase from Enterococcus hirae contains a heptameric rotor. AB - In F-ATPases, ATP hydrolysis is coupled to translocation of ions through membranes by rotation of a ring of c subunits in the membrane. The ring is attached to a central shaft that penetrates the catalytic domain, which has pseudo-3-fold symmetry. The ion translocation pathway lies between the external circumference of the ring and another hydrophobic protein. The H+ or Na+:ATP ratio depends upon the number of ring protomers, each of which has an essential carboxylate involved directly in ion translocation. This number and the ratio differ according to the source, and 10, 11, and 14 protomers have been found in various enzymes, with corresponding calculated H+ or Na+:ATP ratios of 3.3, 3.7, and 4.7. V-ATPases are related in structure and function to F-ATPases. Oligomers of subunit K from the Na+-motive V-ATPase of Enterococcus hirae also form membrane rings but, as reported here, with 7-fold symmetry. Each protomer has one essential carboxylate. Thus, hydrolysis of one ATP provides energy to extrude 2.3 sodium ions. Symmetry mismatch between the catalytic and membrane domains appears to be an intrinsic feature of both V- and F-ATPases. PMID- 12651849 TI - Inhibition of tumor growth with a vaccine based on xenogeneic homologous fibroblast growth factor receptor-1 in mice. AB - Angiogenesis is important for the growth of solid tumors. The breaking of the immune tolerance against the molecule associated with angiogenesis should be a useful approach for cancer therapy. However, the immunity to self-molecules is difficult to elicit by a vaccine based on autologous or syngeneic molecules due to immune tolerance. Basic fibroblast growth factor (bFGF) is a specific and potent angiogenic factor implicated in tumor growth. The biological activity of bFGF is mediated through interaction with its high-affinity receptor, fibroblast growth factor receptor-1 (FGFR-1). In this study, we selected Xenopus FGFR-1 as a model antigen by the breaking of immune tolerance to explore the feasibility of cancer therapy in murine tumor models. We show here that vaccination with Xenopus FGFR-1 (pxFR1) is effective at antitumor immunity in three murine models. FGFR-1 specific autoantibodies in sera of pxFR1-immunized mice could be found in Western blotting analysis. The purified immunoglobulins were effective at the inhibition of endothelial cell proliferation in vitro and at the antitumor activity in vivo. The antitumor activity and production of FGFR-1-specific autoantibodies could be abrogated by depletion of CD4+ T lymphocytes. Histological examination revealed that the autoantibody was deposited on the endothelial cells within tumor tissues from pxFR1-immunized mice, and intratumoral angiogenesis was significantly suppressed. Furthermore, the inhibition of angiogenesis could also be found in alginate-encapsulate tumor cell assay. These observations may provide a new vaccine strategy for cancer therapy through the induction of autoimmunity against FGFR-1 associated with angiogenesis in a cross-reaction. PMID- 12651850 TI - Phosphospecific site tyrosine phosphorylation of p125FAK and proline-rich kinase 2 is differentially regulated by cholecystokinin receptor type A activation in pancreatic acini. AB - The focal adhesion kinases, p125FAK and proline-rich kinase 2 (PYK2), are involved in numerous processes as adhesion, cytoskeletal changes, and growth. These kinases have 45% homology and share three tyrosine phosphorylation (TyrP) sites. Little information exists on the ability of stimulants to cause TyrP of each kinase site and the cellular mechanism involved. We explored the ability of the neurotransmitter/hormone, CCK, to stimulate TyrP at each site. In rat pancreatic acini, CCK stimulated TyrP at each site in both kinases. TyrP was rapid except for pY397FAK. The magnitude of TyrP differed with the different FAK and PYK2 sites. The CCK dose-response curve for TyrP for sites in each kinase was similar. CCK-JMV, an agonist of the high affinity receptor state and antagonist of the low affinity receptor state, was less efficacious than CCK at each FAK/PYK2 site and inhibited CCK maximal stimulation. Thapsigargin decreased CCK stimulated TyrP of pY402PYK2 and pY925FAK but not the other sites. GF109203X reduced TyrP of only the PYK2 sites, pY402 and pY580. GF109203X with thapsigargin decreased TyrP of pY402PYK2 and the three FAK sites more than either inhibitor alone. Basal TyrP of pY397FAK was greater than other sites. These results demonstrate that CCK stimulates tyrosine phosphorylation of each of the three homologous phosphorylation sites in FAK and PYK2. However, CCK-stimulated TyrP at these sites differs in kinetics, magnitude, and participation of the high/low affinity receptor states and by protein kinase C and [Ca2+]i. These results show that phosphorylation of these different sites is differentially regulated and involves different intracellular mechanisms in the same cell. PMID- 12651851 TI - Structural basis for flip-flop action of thiamin pyrophosphate-dependent enzymes revealed by human pyruvate dehydrogenase. AB - The derivative of vitamin B1, thiamin pyrophosphate, is a cofactor of enzymes performing catalysis in pathways of energy production. In alpha2beta2 heterotetrameric human pyruvate dehydrogenase, this cofactor is used to cleave the Calpha-C(=O) bond of pyruvate followed by reductive acetyl transfer to lipoyl dihydrolipoamide acetyltransferase. The dynamic nonequivalence of two, otherwise chemically equivalent, catalytic sites has not yet been understood. To understand the mechanism of action of this enzyme, we determined the crystal structure of the holo-form of human pyruvate dehydrogenase at 1.95-A resolution. We propose a model for the flip-flop action of this enzyme through a concerted approximately 2 A shuttle-like motion of its heterodimers. Similarity of thiamin pyrophosphate binding in human pyruvate dehydrogenase with functionally related enzymes suggests that this newly defined shuttle-like motion of domains is common to the family of thiamin pyrophosphate-dependent enzymes. PMID- 12651852 TI - Insulin-like growth factor-1/insulin bypasses Pref-1/FA1-mediated inhibition of adipocyte differentiation. AB - Pref-1 is a highly glycosylated Delta-like transmembrane protein containing six epidermal growth factor-like repeats in the extracellular domain. Pref-1 is abundantly expressed in preadipocytes, but expression is down-regulated during adipocyte differentiation. Forced expression of Pref-1 in 3T3-L1 cells was reported to inhibit adipocyte differentiation. Here we show that efficient and regulated processing of Pref-1 occurs in 3T3-L1 preadipocytes releasing most of the extracellular domain as a 50-kDa heterogeneous protein, previously isolated and characterized as FA1. Unexpectedly, we found that forced expression of the soluble form, FA1, or full-length Pref-1 did not inhibit adipocyte differentiation of 3T3-L1 cells when differentiation was induced by standard treatment with methylisobutylxanthine, dexamethasone, and high concentrations of insulin. However, forced expression of either form of Pref-1/FA1 in 3T3-L1 or 3T3 F442A cells inhibited adipocyte differentiation when insulin or insulin-like growth factor-1 (IGF-1) was omitted from the differentiation mixture. We demonstrate that the level of the mature form of the IGF-1 receptor is reduced and that IGF-1-dependent activation of p42/p44 mitogen-activated protein kinases (MAPKs) is compromised in preadipocytes with forced expression of Pref-1. This is accompanied by suppression of clonal expansion and terminal differentiation. Accordingly, supplementation with insulin or IGF-1 rescued p42/p44 MAPK activation, clonal expansion, and adipocyte differentiation in a dose-dependent manner. PMID- 12651853 TI - Syntaxin isoform specificity in the regulation of renal H+-ATPase exocytosis. AB - Intercalated and inner medullary collecting duct (IMCD) cells of the kidney mediate the transport of H+ by a plasma membrane H+-ATPase. The rate of H+ transport in these cells is regulated by exocytic insertion of H+-ATPase-laden vesicles into the apical membrane. We have shown that the exocytic insertion of proton pumps (H+-ATPase) into the apical membrane of rat IMCD cells, in culture, involves SNARE proteins (syntaxin (synt), SNAP-23, and VAMP). The membrane fusion complex observed in IMCD cells with the induction of proton pump exocytosis not only included these SNAREs but also the H+-ATPase. Based on these observations, we suggested that the targeting of these vesicles to the apical membrane is mediated by an interaction between the H+-ATPase and a specific t-SNARE. To evaluate this hypothesis, we utilized a "pull-down" assay in which we identified, by Western analysis, the proteins in a rat kidney medullary homogenate that complexed with glutathione S-transferase (GST) fusion syntaxin isoforms attached to Sepharose 4B-glutathione beads. The syntaxin isoforms employed were 1A, 1B, 2, 4, 5, and also 1A that was truncated to exclude the H3 SNARE binding domain (synt 1ADeltaH3). All full-length syntaxin isoforms formed complexes with SNAP-23 and VAMP. Neither GST nor synt-1ADeltaH3 formed complexes with these SNAREs. H+ ATPase (subunits E, a, and c) bound to syntaxin-1A and to a lesser extent to synt 1B but not to synt-1ADeltaH3 or synt-2, -4, and -5. In cultured IMCD cells transfected to express syntaxin truncated for the membrane binding domain (synt DeltaC), expression of synt-1ADeltaC, but not synt-4DeltaC, inhibited H+-ATPase exocytosis. In conclusion, because all full-length syntaxins examined bound VAMP 2 and SNAP-23, but only non-H3-truncated syntaxin-1 bound H+-ATPase, and synt 1ADeltaC expression by intact IMCD cells inhibited H+-ATPase exocytosis, it is likely that the H+-ATPase binds directly to the H3 domain of syntaxin-1 and not through VAMP-2 or SNAP-23. Interaction between the syntaxin-1A and H+-ATPase is important in the targeted exocytosis of the proton pump to the apical membrane of intercalated cells. PMID- 12651854 TI - Binding and internalization of lipopolysaccharide by Cla-1, a human orthologue of rodent scavenger receptor B1. AB - Scavenger receptor, class B, type I (SR-BI) mediates selective uptake of high density lipoprotein (HDL) cholesteryl ester. SR-BI recognizes HDL, low density lipoprotein (LDL), exchangeable apolipoproteins, and protein-free lipid vesicles containing negatively charged phospholipids. Lipopolysaccharides (LPS) are highly glycosylated anionic phospholipids contributing to septic shock. Despite significant structural similarities between anionic phospholipids and LPS, the role of SR-BI in LPS uptake is unknown. Cla-1, the human SR-BI orthologue, was determined to be a LPS-binding protein and endocytic receptor mediating the binding and internalization of lipoprotein-free, monomerized LPS. LPS strongly competed with HDL, lipidfree apoA-I and apoA-II for HDL binding to the mouse RAW cells. Stably transfected HeLa cells expressing Cla-1-bound LPS with a Kd of about 16 microg/ml, and had a 3-4-fold increase in binding capacity and LPS uptake. Bodipy-labeled LPS uptake was found to initially accumulate in the plasma membrane and subsequently in a perinuclear region identified predominantly as the Golgi complex. Bodipy-LPS and Alexa-apoA-I had staining that colocalized on the cell surface and intracellularly indicating similar transport mechanisms. When associated with HDL, LPS uptake was increased in Cla-1 overexpressing HeLa cells by 5-10-fold. Cla-1-associated 3H-LPS uptake exceeded 125I-apolipoprotein uptake by 5-fold indicating a selective LPS uptake. Upon interacting with Cla-1 overexpressing HeLa cells, the complex (Bodipy-LPS/Alexa 488 apolipoprotein labeled HDL) bound and was internalized as a holoparticle. Intracellularly, LPS and apolipoproteins were sorted to different intracellular compartments. With LPS associated HDL, intracellular LPS co-localized predominantly with transferrin, indicating delivery to an endocytic recycling compartment. Our study reveals a close similarity between Cla-1-mediated selective LPS uptake and the recently described selective lipid sorting by rodent SR-BI. In summary, Cla-1 was found to bind and internalize monomerized and HDL-associated LPS, indicating that Cla-1 may play important role in septic shock by affecting LPS cellular uptake and clearance. PMID- 12651855 TI - Participation of a fusogenic protein, glyceraldehyde-3-phosphate dehydrogenase, in nuclear membrane assembly. AB - We found an autoimmune serum, K199, that strongly suppresses nuclear membrane assembly in a cell-free system involving a Xenopus egg extract. Four different antibodies that suppress nuclear assembly were affinity-purified from the serum using Xenopus egg cytosol proteins. Three proteins recognized by these antibodies were identified by partial amino acid sequencing to be glyceraldehyde-3-phosphate dehydrogenase (GAPDH), fructose-1,6-bisphosphate aldolase, and the regulator of chromatin condensation 1. GAPDH is known to be a fusogenic protein. To verify the participation of GAPDH in nuclear membrane fusion, authentic antibodies against human and rat GAPDH were applied, and strong suppression of nuclear assembly at the nuclear membrane fusion step was observed. The nuclear assembly activity suppressed by antibodies was recovered on the addition of purified chicken GAPDH. A peptide with the sequence of amino acid residues 70-94 of GAPDH, which inhibits GAPDH-induced phospholipid vesicle fusion, inhibited nuclear assembly at the nuclear membrane fusion step. We propose that GAPDH plays a crucial role in the membrane fusion step in nuclear assembly in a Xenopus egg extract cell-free system. PMID- 12651856 TI - Characterization of regulatory elements and methylation pattern of the autoimmune regulator (AIRE) promoter. AB - Defects in the AIRE gene cause a monogenic autoimmune syndrome APECED (autoimmune polyendocrinopathy candidiasis ectodermal dystrophy), which is characterized by loss of self-tolerance to multiple organs. In concordance with its role in immune tolerance, AIRE is most strongly expressed in thymic epithelial cells and in cells of monocytic-dendritic lineage. The AIRE protein has been shown to function as a transcriptional regulator, however, the mechanisms regulating AIRE gene expression are not known. Here we have characterized the AIRE promoter region by identifying a minimal promoter region within 350 bp of the translation initiation codon. Electrophoretic mobility shift assays and transient transfections with mutated promoter constructs revealed a functional TATA box (-163 to -153) and binding sites for transcription complexes AP-1 (-307 to -296), NF-Y (-213 to 202), and Sp1 (-202 to -189). The presence of a 390-bp CpG island within the proximal promoter suggested that cytosine methylation has a role in transcriptional regulation of AIRE, which was supported by in vitro methylation experiments of promoter constructs. Sodium bisulfite sequencing showed a less methylated status of AIRE promoter in the thymic epithelial cell line TEC1A3 compared with HeLa and monocytic cells U937 and THP-1. Real-time PCR analysis showed that treatment with 5-aza-2'-deoxycytidine (5-azaCdR), a DNA methyltransferase inhibitor, up-regulated AIRE transcript levels in TEC1A3, U937, and HeLa cells and that even greater activations in TEC1A3 and U937 cells were observed using combined treatments with deacetylase inhibitor trichostatin A. These results suggest that AIRE gene expression is modulated through modifications in chromatin methylation and acetylation. PMID- 12651857 TI - HspBP1, an Hsp70 cochaperone, has two structural domains and is capable of altering the conformation of the Hsp70 ATPase domain. AB - We present here the first structural information for HspBP1, an Hsp70 cochaperone. Using circular dichroism, HspBP1 was determined to be 35% helical. Although HspBP1 is encoded by seven exons, limited proteolysis shows that it has only two structural domains. Domain I, amino acids 1-83, is largely unstructured. Domain II, amino acids 84-359, is predicted to be 43% helical using circular dichroism. Using limited proteolysis we have also shown that HspBP1 association changes the conformation of the ATPase domain of Hsp70. Only domain II of HspBP1 is required to bring about this conformational change. Truncation mutants of HspBP1 were tested for their ability to inhibit the renaturation of luciferase and bind to Hsp70 in reticulocyte lysate. A carboxyl terminal truncation mutant that was slightly longer than domain I was inactive in these assays, but domain II was sufficient to perform both functions. Domain II was less active than full length HspBP1 in these assays, and addition of amino acids from domain I improved both functions. These studies show that HspBP1 domain II can bind Hsp70, change the conformation of the ATPase domain, and inhibit Hsp70-associated protein folding. PMID- 12651858 TI - The role of the C terminus and Na+/H+ exchanger regulatory factor in the functional expression of cystic fibrosis transmembrane conductance regulator in nonpolarized cells and epithelia. AB - The conserved C-terminal peptide motif (1476DTRL) of the cystic fibrosis transmembrane conductance regulator (CFTR) ensures high affinity binding to different PSD-95/Disc-large/zonula occludens-1 (PDZ) domain-containing molecules, including the Na+/H+ exchanger regulatory factor (NHERF)/ezrin-radixin-moesin binding phosphoprotein of 50 kDa. The physiological relevance of NHERF binding to CFTR is not fully understood. Individuals with mutations resulting in premature termination of CFTR (S1455X or Delta26 CFTR) have moderately elevated sweat Cl- concentration, without an obvious lung and pancreatic phenotype, implying that the CFTR function is largely preserved. Surprisingly, when expressed heterologously, the Delta26 mutation was reported to abrogate channel activity by destabilizing the protein at the apical domain and inducing its accumulation at the basolateral membrane (Moyer, B., Denton, J., Karlson, K., Reynolds, D., Wang, S., Mickle, J., Milewski, M., Cutting, G., Guggino, W., Li, M., and Stanton, B. (1999) J. Clin. Invest. 104, 1353-1361). The goals of this study were to resolve the contrasting clinical and cellular phenotype of the Delta26 CFTR mutation and evaluate the role of NHERF in the functional expression of CFTR at the plasma membrane. Complex formation between CFTR and NHERF was disrupted by C-terminal deletions, C-terminal epitope tag attachments, or overexpression of a dominant negative NHERF mutant. These perturbations did not alter CFTR expression, metabolic stability, or function in nonpolarized cells. Likewise, inhibition of NHERF binding had no discernible effect on the apical localization of CFTR in polarized tracheal, pancreatic, intestinal, and kidney epithelia and did not influence the metabolic stability or the cAMP-dependent protein kinase-activated chloride channel conductance in polarized pancreatic epithelia. On the other hand, electrophysiological studies demonstrated that NHERF is able to stimulate the cAMP-dependent protein kinase-phosphorylated CFTR channel activity in intact cells. These results help to reconcile the discordant genotype-phenotype relationship in individuals with C-terminal truncations and indicate that apical localization of CFTR involves sorting signals other than the C-terminal 26 amino acid residues and the PDZ-binding motif in differentiated epithelia. PMID- 12651859 TI - Dioxolane guanosine 5'-triphosphate, an alternative substrate inhibitor of wild type and mutant HIV-1 reverse transcriptase. Steady state and pre-steady state kinetic analyses. AB - The frequency of human immunodeficiency virus, type 1 (HIV-1) mutations in response to antiviral therapy and resulting drug resistance is of major concern. Amdoxovir ((-)-beta-D-2,6-diaminopurine dioxolane), the prodrug of dioxolane guanosine (DXG), is currently in phase I/II clinical development for the treatment of HIV-1 infection. In vitro, HIV-1 mutants resistant to 3'-azido-3' deoxythymidine (M41L/D67N/K70R/T215Y/K219Q) and (-)beta-L-2',3'-dideoxy-3' thiacytidine (3TC) (M184V) remain sensitive to DXG. HIV-1 with the reverse transcriptase mutations K65R, L74V, and/or Q151M were less sensitive to DXG, whereas the mutation K103N re-sensitized the virus to the inhibitory effect of DXG. In order to understand these observations at the enzyme level, we investigated the inhibition of the HIV-1 reverse transcriptase-catalyzed viral DNA synthesis by dioxolane guanosine 5'-triphosphate (DXG-TP), 3'-azido-3' deoxythymidine-TP, and 3TC-TP by using steady state kinetic analysis and the incorporation of DXG-5'-monophosphate by using pre-steady state kinetic analysis. This mechanistic study provided detailed information on the amdoxovir-related drug resistance at a molecular level. Overall, the enzymatic data correlated well with the antiviral data obtained from cell culture experiments and further supported the use of amdoxovir for the treatment of nucleoside reverse transcriptase inhibitor-experienced patients. PMID- 12651860 TI - The cohesin SMC3 is a target the for beta-catenin/TCF4 transactivation pathway. AB - The structural maintenance of chromosome protein SMC3 is a component of the cohesin complex that mediates sister chromatid cohesion and segregation in prokaryotes and eukaryotes. It is also present extracellularly in the form of a chondroitin sulfate proteoglycan known as bamacan. We have found previously that SMC3 expression is elevated in a large fraction of human colon carcinomas. The additional finding that the protein is significantly increased in the intestinal polyps of ApcMin/+ mice has led us to hypothesize that SMC3 expression is linked to activation of the APC/beta-catenin/TCF4 pathway. The immunohistochemical analysis of colon adenocarcinomas from clinical specimens revealed that beta catenin and SMC3 antigens co-localize with maximal stain intensity within the transformed areas. Cloning and sequencing of 1578 bp of the human SMC3 promoter unveiled the presence of seven putative consensus sequences for beta-catenin/TCF4 binding, two of which are conserved in the mouse Smc3 promoter. Transient transfection experiments in HCT116 and SW480 human colon carcinoma cells using deletion and mutated promoter constructs in luciferase reporter vectors confirmed that the putative sites, the first located at -48 bp and the second located at 701 bp, are susceptible to beta-catenin/TCF4 transactivation. Co-transfection with a beta-catenin expression vector enhanced the promoter activity whereas E cadherin had the opposite effect. Binding of beta-catenin/TCF4 complexes from SW480 nuclear extracts to these sequences was confirmed by electrophoretic shift and supershift mobility assays. Altogether these results are consistent with the idea that the beta-catenin/TCF4 transactivation pathway contributes to SMC3 overexpression in intestinal tumorigenesis. PMID- 12651861 TI - Fluctuations in acceleration during voluntary contractions lead to greater impairment of movement accuracy in old adults. AB - The purpose of the study was to assess the effect of movement velocity on the relation between fluctuations in acceleration and the ability to achieve a target velocity during voluntary contractions performed by young (29.5 +/- 4.3 yr) and old (74.9 +/- 6.2 yr) adults. Subjects performed concentric and eccentric contractions with the first dorsal interosseus muscle while lifting a submaximal load (15% of maximum) at six movement velocities (0.03-1.16 rad/s). Fluctuations in acceleration, the accuracy of matching the target velocity, and electromyographic (EMG) activity were determined from three trials for each contraction type and movement velocity. The fluctuations in acceleration increased with movement velocity for both concentric and eccentric contractions, but they were greatest during fast eccentric contractions ( approximately 135%) when there was stronger modulation of acceleration in the 5- to 10-Hz bandwidth. Nonetheless, EMG amplitude for first dorsal interosseus increased with movement velocity only for concentric and not eccentric contractions. Consistent with the minimum variance theory, movement accuracy was related to the fluctuations in acceleration for both types of contractions in all subjects. For a given level of fluctuations in acceleration, however, old subjects were three times less accurate than young subjects. Although the EMG amplitude at each speed was similar for young and old adults, only the young adults modulated the power in the EMG spectrum with speed. Thus the fluctuations in acceleration during voluntary contractions had a more pronounced effect on movement accuracy for old adults compared with young adults, probably due to factors that influenced the frequency-domain characteristics of the EMG. PMID- 12651862 TI - Effects of aerobic training on heart rate dynamics in sedentary subjects. AB - This study was designed to assess the effects of moderate- and high-volume aerobic training on the time domain and on spectral and fractal heart rate (HR) variability indexes. Sedentary subjects were randomized into groups with moderate volume training (n = 20), high-volume training (n = 20), and controls (n = 15). The training period was 8 wk, including 6 sessions/wk at an intensity of 70-80% of the maximum HR, lasting for 30 min/session in the moderate-volume group and 60 min/session in the high-volume group. Time domain, frequency domain, and short term fractal scaling measures of HR variability were analyzed over a 24-h period. Mean HR decreased from 70 +/- 7 to 64 +/- 8 beats/min and from 67 +/- 5 to 60 +/- 6 beats/min (P < 0.001 for both) for the moderate- and high-volume training groups, respectively. The normalized high-frequency spectral component increased in both groups (P < 0.05). The normalized low-frequency component decreased significantly (P < 0.05), resulting in a marked decrease in low frequency-to-high frequency ratio in both groups. In addition, short-term scaling exponent decreased in both groups (P < 0.001). There were no significant differences in the changes of HR variability indexes between groups. Aerobic training in sedentary subjects results in altered autonomic regulation of HR toward vagal dominance. A moderate training volume is a sufficient intervention to induce these beneficial effects. PMID- 12651863 TI - B-1 cell (CD5+/CD11b+) numbers and nIgM levels are elevated in physically active vs. sedentary rats. AB - Habitual, moderate exercise is associated with improved health, including reductions in illness. These benefits may stem, in part, from immune function improvements. We have previously reported that daily wheel running increases serum and peritoneal natural IgM (nIgM) in pathogen-free Sprague-Dawely rats. B-1 cells, which primarily reside in the peritoneal cavity, produce nIgM in the absence of antigen stimulation. This study examined whether physical activity would also increase B-1 cell numbers in the peritoneal cavity, mesenteric lymph nodes, and spleen. Male, pathogen-free Fischer 344 rats were sedentary (standard cages) or physically active (running wheel access) for 6-7 wk. Peritoneal cavity, mesenteric lymph nodes, and spleen cells were taken, and the number of CD5+/CD11b+ (B-1) cells were measured by using two-color flow cytometry. The results were that physically active animals had increased numbers of CD5+/CD11b+ cells in the peritoneal cavity. In addition, physically active animals had increased serum and peritoneal nIgM, thus replicating our previous observations. These results indicate that voluntary running selectively increases the B-1 cell population, which is most likely responsible for the elevated serum and peritoneal nIgM in active rats. Because B-1 cells are important in host defense, these changes may contribute to the health benefits of exercise. PMID- 12651864 TI - Modeling airflow-related shear stress during heterogeneous constriction and mechanical ventilation. AB - Ventilator-induced lung injury has been proposed as being caused by overdistention and closure and reopening of small airways and alveoli. Here we investigate the possibility that heterogeneous constriction increases airflow related shear stress to a dangerously high level that may be sufficient to cause injury to the epithelial cells during mechanical ventilation. We employed an anatomically consistent model of the respiratory system, based on Horsfield morphometric data, and solved for the time evolution of pressure and flow along the airway tree during mechanical ventilation. We simulated constant-flow ventilation with passive expiration in two different conditions: baseline and highly heterogeneous constriction. The constriction was applied with two strategies: establishing a simple diameter reduction or adding also a length shortening. The shear stress distribution on airway walls was analyzed for airways ranging from the trachea to the acini. Our results indicate that 1). heterogeneous constriction can amplify the maximal values of shear stress up to 50-fold, with peak values higher than 0.6 cmH2O; 2). the highest shear stress is found in pathways constricted by 60-80%; 3). simultaneous diameter reduction and shortening amplifies the shear stresses by three- to fourfold, with shear stresses reaching 2 cmH2O; and 4). there is a range of airways (diameters from 0.6 to 0.3 mm at baseline) that appear to be at risk of very high stresses. We conclude that elevated airflow-related shear stress on the epithelial cell layer can occur during heterogeneous constriction and conjecture that this may constitute a mechanism contributing to ventilator-induced lung injury. PMID- 12651866 TI - Comparison of the genomes of human and mouse lays the foundation of genome zoology. AB - The extensive similarities between the genomes of human and model organisms are the foundation of much of modern biology, with model organism experimentation permitting valuable insights into biological function and the aetiology of human disease. In contrast, differences among genomes have received less attention. Yet these can be expected to govern the physiological and morphological distinctions apparent among species, especially if such differences are the result of evolutionary adaptation. A recent comparison of the draft sequences of mouse and human genomes has shed light on the selective forces that have predominated in their recent evolutionary histories. In particular, mouse-specific clusters of homologues associated with roles in reproduction, immunity and host defence appear to be under diversifying positive selective pressure, as indicated by high ratios of non-synonymous to synonymous substitution rates. These clusters are also frequently punctuated by homologous pseudogenes. They thus have experienced numerous gene death, as well as gene birth, events. These regions appear, therefore, to have borne the brunt of adaptive evolution that underlies physiological and behavioural innovation in mice. We predict that the availability of numerous animal genomes will give rise to a new field of genome zoology in which differences in animal physiology and ethology are illuminated by the study of genomic sequence variations. PMID- 12651867 TI - PQBP-1 transgenic mice show a late-onset motor neuron disease-like phenotype. AB - A body of experimental evidence indicates that transcription and/or mRNA processing factors interacting with the polyglutamine disease gene products play crucial roles in the pathology. PQBP-1 is one of these factors and it has been shown to interact with the spinocerebellar ataxia type-1 (SCA1) disease gene product, ataxin-1. Our previous data suggested that relatively high expression of PQBP-1 in the cerebellum might explain the selective neuronal degeneration of SCA1. To further test whether PQBP-1 expression level regulates neuronal death, we generated transgenic mice of human PQBP-1 driven by a regulatory element for ubiquitous gene expression. The mice showed a late-onset and gradually progressive motor neuron disease-like phenotype, which might be related to neurogenic muscular atrophy observed in SCA1 patients. Ataxia could not be discriminated from predominant progressive weakness. Pathological examinations of the transgenic mice revealed loss of Purkinje and granular cells in the cerebellum as well as that of spinal motor neurons, corresponding to the pathology of human SCA1. These findings show that excessive action of PQBP-1 causes neuronal dysfunction and support PQBP-1 being involved in the pathology of SCA1. PMID- 12651868 TI - Defective secretion of recombinant fragments of fibrillin-1: implications of protein misfolding for the pathogenesis of Marfan syndrome and related disorders. AB - Fibrillin-1 is a large modular glycoprotein that assembles to form 10-12 nm microfibrils in the extracellular matrix. Mutations in the fibrillin-1 gene (FBN1) cause Marfan syndrome and related connective tissue disorders (fibrillinopathies) that show autosomal dominant inheritance. The pathogenic mechanism is thought to be a dominant negative effect of a mutant protein on microfibril assembly, although direct evidence is lacking. A significant group of disease-causing FBN1 mutations are cysteine substitutions within EGF domains that are predicted to cause misfolding by removal of disulphide bonds that stabilize the native domain fold. We have studied three missense mutations (C1117Y, C1129Y and G1127S) to investigate the effect of misfolding on the trafficking of fibrillin-1 from fibroblast cells. We demonstrate that both C1117Y and C1129Y, expressed as recombinant fragments of fibrillin-1, are retained and accumulate within the cell. Both undergo core glycosylation but lack the complex glycosylation observed in the secreted wild-type fragment, suggesting retention in the endoplasmic reticulum (ER). In addition, co-immunoprecipitation experiments show association with the ER chaperone calreticulin, but not calnexin, 78 kDa glucose-regulated protein (Grp78/BiP) or protein disulfide isomerase. In contrast, G1127S, which causes a moderate change in the EGF domain fold, shows a pattern of glycosylation and trafficking profile indistinguishable from the wild-type fragment. Since expression of the recombinant fragments does not disrupt the secretion of endogenous fibrillin-1 by the cell, we propose that G1127S causes disease via an extracellular dominant negative effect. In contrast, the observed ER retention of C1117Y and C1129Y suggests that disease associated with these missense mutations is caused either by an intracellular dominant negative effect or haploinsufficiency. PMID- 12651870 TI - Aggresomes protect cells by enhancing the degradation of toxic polyglutamine containing protein. AB - Expression of misfolded protein in cultured cells frequently leads to the formation of juxtanuclear inclusions that have been termed 'aggresomes'. Aggresome formation is an active cellular response that involves trafficking of the offending protein along microtubules, reorganization of intermediate filaments and recruitment of components of the ubiquitin proteasome system. Whether aggresomes are benevolent or noxious is unknown, but they are of particular interest because of the appearance of similar inclusions in protein deposition diseases. Here we present evidence that aggresomes serve a cytoprotective function and are associated with accelerated turnover of mutant proteins. We show that mutant androgen receptor (AR), the protein responsible for X-linked spinobulbar muscular atrophy, forms insoluble aggregates and is toxic to cultured cells. Mutant AR was also found to form aggresomes in a process distinct from aggregation. Molecular and pharmacological interventions were used to disrupt aggresome formation, revealing their cytoprotective function. Aggresome forming proteins were found to have an accelerated rate of turnover, and this turnover was slowed by inhibition of aggresome formation. Finally, we show that aggresome-forming proteins become membrane-bound and associate with lysosomal structures. Together, these findings suggest that aggresomes are cytoprotective, serving as cytoplasmic recruitment centers to facilitate degradation of toxic proteins. PMID- 12651869 TI - E-box mutations in the RAPSN promoter region in eight cases with congenital myasthenic syndrome. AB - Myogenic determination factors are basic helix-loop-helix proteins that govern specification and differentiation of muscle cells, and bind to the E-box consensus sequence CANNTG in promoter regions of muscle-specific genes. No E-box mutation has been reported to date. RAPSN encodes rapsyn, a 43 kDa postsynaptic peripheral membrane protein that clusters the nicotinic acetylcholine receptor at the motor endplate. Transcriptional regulation mechanisms of RAPSN have not been studied. We here report two novel E-box mutations in the RAPSN promoter region in eight congenital myasthenic syndrome patients. Patient 1 carries -27C-->G that changes an E-box at -27 to -22 from CAGCTG to GAGCTG. An allele harboring -27C- >G is not transcribed in patient's muscle. Patients 2-8 are of Oriental Jewish stock of Iraqi or Iranian origin with facial malformations, and harbor -38A-->G that changes another E-box at -40 to -35 from CAACTG to CAGCTG, which does not affect the consensus CANNTG sequence. Haplotype analysis shows that -38A-->G arises from a common founder. For each mutation, position +1 represents the major transcriptional start site that we determine to be 172 nucleotides upstream of the translational start site. Electrophoretic mobility shift assays reveal that 38A-->G gains, and -27C-->G looses, binding affinity for different components of nuclear extracts of C2C12 myotubes. Luciferase reporter assays show that both 38A-->G and -27C-->G attenuate reporter gene expression in C2C12 myotubes, and that -27C-->G additionally attenuates reporter gene expression in MyoD- or myogenin-transfected HEK cells. The -27C-->G mutation also markedly attenuates the enhancer activity of an E-box on an SV40 promoter. Impaired transcriptional activities of the RAPSN promoter region predict reduced rapsyn expression and endplate acetylcholine receptor deficiency. PMID- 12651871 TI - Alternative 5' exons of the CFTR gene show developmental regulation. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) gene shows a complex mechanism of tissue-specific and temporal regulation. Expression of the sheep and human CFTR genes shows a gradual decline during lung development, from the early mid-trimester through to term. Alternative upstream exons of CFTR have been identified in several species but their functional role remains obscure. We identified a novel 5' exon of the sheep CFTR gene (ov1a) that occurs in two splice forms (ov1aL and ov1aS), which are both mutually exclusive with exon 1. CFTR transcripts including ov1aL and ov1aS are present at low levels in many sheep tissues, however ov1aS shows temporal and spatial regulation during fetal lung development, being most abundant when CFTR expression levels start to decline. Alternative 5' exons -1a and 1a in the human CFTR gene also show changes in expression levels through lung development. Evaluation of ov1aL and ov1aS by Mfold reveals the potential to form extremely stable secondary structures which would cause ribosomal subunit detachment. Further, the loss of exon 1 from the CFTR transcript removes motifs that are crucial for normal trafficking of the CFTR protein. Recruitment of these alternative upstream exons may represent a novel mechanism of developmental regulation of CFTR expression. PMID- 12651872 TI - Linkage disequilibrium patterns of the human genome across populations. AB - We studied the patterns of linkage disequilibrium (LD) in the human genome among three populations: African Americans, Caucasians and Ashkenazi Jews. These three populations represent admixed, outbred and isolated populations, respectively. The study examined defined chromosomal regions across the whole genome. We found that SNP allele frequencies are highly correlated between Ashkenazi Jews and Caucasians and somewhat distinct in African Americans. In addition, Ashkenazi Jews have a modest increase in LD compared with Caucasians, and both have greater LD than African Americans. The three populations differed more significantly with regard to haplotype heterogeneity. We found, as expected, that Ashkenazi Jews display the greatest extent of homogeneity and African Americans the greatest extent of heterogeneity. We found that most of the variance in LD can be attributed to the difference between regions and markers rather than to that between different population types. The average recombination rates estimated by low-resolution genetic maps can only explain a small fraction of the variance between regions. We found that LD (in terms of r(2)) decreases as a function of distance even within the so-called 'haplotype blocks'. This has significant consequences when using LD mapping for the genetic dissection of complex traits, as higher density SNP maps will be required to scan the genome. PMID- 12651873 TI - Non-syndromic vestibular disorder with otoconial agenesis in tilted/mergulhador mice caused by mutations in otopetrin 1. AB - Otoconia are biominerals within the utricle and saccule of the inner ear that are critical for the perception of gravity and linear acceleration. The classical mouse mutant tilted (tlt) and a new allele, mergulhador (mlh), are recessive mutations that affect balance by impairing otoconial morphogenesis without causing collateral deafness. The mechanisms governing otoconial biosynthesis are not known. Here we show that tlt and mlh are mutant alleles of a novel gene (Otopetrin 1, Otop1), encoding a multi-transmembrane domain protein that is expressed in the macula of the developing otocyst. Both mutants carry single point mutations leading to non-conservative amino acid substitutions that affect two putative transmembrane (TM) domains (tlt, Ala(151)-->Glu in TM3; mlh, Leu(408)-->Gln in TM8). Otop1 and Otop1-like paralogues, Otop2 and Otop3, define a new gene family with homology to the C. elegans and D. melanoganster DUF270 genes. PMID- 12651874 TI - A comprehensive search for DNA amplification in lung cancer identifies inhibitors of apoptosis cIAP1 and cIAP2 as candidate oncogenes. AB - Amplification of oncogenes is an important mechanism that can cause gene overexpression and contributes to tumor development. The identification of amplified regions might have both prognostic and therapeutic significance. We used primary lung carcinomas and lung cancer cell lines for restriction landmark genomic scanning (RLGS) to identify novel amplified sequences. Enhanced RLGS fragments that indicate gene amplification were observed in primary tumors and lung cancer cell lines of both non-small cell lung cancer and small cell lung cancer. We identified one novel amplicon on chromosome 11q22, in addition to previously reported amplicons that include oncogenes MYCC, MYCL1 and previously identified amplification of chromosomal regions 6q21 and 3q26-27. Amplification of 11q22 has been reported in other types of cancer and was refined to an approximately 1.19 Mbp region for which the complete sequence is available. Based on a patient sample with a small region of low-level amplification we were able to further narrow this region to 0.92 Mbp. Genes localized in this region include two inhibitors of apoptosis (cIAP1 and cIAP2). Immunohistochemistry and western blot analysis identified cIAP1 and cIAP2 as potential oncogenes in this region as both are overexpressed in multiple lung cancers with or without higher copy numbers. PMID- 12651875 TI - Irruption of genomics in the search for disease related genes. AB - Genomics was initiated when robotics made possible the characterisation of large numbers of DNA fragments and when ever improving computers with dedicated software were applied to the localisation in the genome of these sequences and to the analysis of their content. By enabling the generation and management of large amounts of DNA based sequences these tools have changed our perception of the genomes of living organisms. These data, as applied to humans, are contributing to the understanding of gene function, disease processes, and evolution of our species. Presently they are changing the research strategies for identifying genetic variations influencing disease susceptibility and response to treatment. These advances will have a profound impact in biomedicine. PMID- 12651876 TI - How to identify the genetic basis of gastrointestinal and liver diseases? AB - New insights into the genetic basis of disease are being generated at an ever increasing rate. This explosion of information was ignited by technological advances, such as the polymerase chain reaction and automated DNA sequencing. Although its promise is great, the integration of genetics into the everyday practice of medicine remains challenging. This review discusses the application of molecular genetics in general with a specific focus on hereditary diseases of the digestive organs. The application of molecular genetics in everyday clinical routine is hampered by the difficult interpretation of test results. These difficulties include the prediction of disease penetrance, the presence of multiple mutations of a particular gene with varying functional consequences, and the importance of exogenous factors modulating disease expression. To date, the most significant impact of genetics has been to increase our understanding of disease aetiology and pathogenesis and to reliably identify siblings of affected patients with the risk to develop symptomatic disease. PMID- 12651878 TI - Genetic testing for polyposis: practical and ethical aspects. AB - The three autosomal dominant inherited polyposis syndromes, familial adenomatous polyposis, juvenile polyposis, and Peutz-Jeghers polyposis predispose to colorectal cancer as does hereditary non-polyposis colorectal cancer syndrome. Uncovering the genetic background of these four cancer traits provides the possibility for genetic testing of the family members of an affected patient. Before testing identification of the underlying family specific pathogenic mutation is mandatory. This is possible in about 60% to 95% of families. Endoscopic surveillance can be safely discontinued in mutation negative family members and surveillance or prophylactic surgery can be targeted to mutation positive members alone. Testing requires genetic counselling and written informed consent to prevent misunderstanding and to minimise untoward effects such as anxiety. Permanent surveillance and adequate prophylactic treatment for all mutation positive subjects and families is best ensured in national or regional polyposis registries with the capacity to take care of long term follow up from generation to generation. PMID- 12651879 TI - Haemochromatosis. AB - Iron is an important component of the Earth's crust, but its own chemistry greatly limits utilisation and also sets the basis for its toxicity. The capacity of readily exchanging electrons in aerobic conditions makes iron essential for fundamental cell functions, such as DNA synthesis, transport of oxygen and electrons, and cell respiration. On the other hand, as humans have no means to control iron excretion, excess iron, regardless of the route of entry, accumulates in parenchymal organs and threatens cell viability. In fact, a number of disease states (that is, iron overload diseases) attributable to genetic or acquired factors are pathogenetically linked to excess body iron stores and iron removal therapy is an effective lifesaving strategy in such circumstances. PMID- 12651877 TI - Pharmacogenomics: marshalling the human genome to individualise drug therapy. AB - Pharmacogenomics aims to identify the inherited basis for interindividual differences in drug response, and translate this to molecular diagnostics that can be used to individualise drug therapy. This review uses a number of published examples of inherited differences in drug metabolising enzymes, drug transporters, and drug targets (for example, receptors) to illustrate the potential importance of inheritance in determining the efficacy and toxicity of medications in humans. It seems that this field is at the early stages of developing a powerful set of molecular diagnostics that will have profound utility in optimising drug therapy for individual patients. PMID- 12651880 TI - Chronic pancreatitis and cystic fibrosis. AB - Recent discoveries of trypsinogen and trypsin inhibitor mutations in patients with chronic pancreatitis (CP) support the hypothesis that an inappropriate activation of pancreatic zymogens to active enzymes within the pancreatic parenchyma starts the inflammatory process. Current data suggest that CP may be inherited dominant, recessive, or complex as a result of mutations in the above mentioned or yet unidentified genes. Evaluation of patients with CP should include genetic testing. Cystic fibrosis (CF) is an autosomal recessive inherited disorder caused by mutations in the CF transmembrane conductance regulator (CFTR) gene and is characterised by pancreatic insufficiency and chronic bronchopulmonary infection. The progression and severity of pulmonary disease differs considerably between people with identical CFTR mutations and does not seem to correlate with the type or class of the CFTR mutation. The identification of further disease modifying genetic factors will increase the pathophysiological understanding and may help to identify new therapeutic targets. PMID- 12651882 TI - The promise of gene therapy in gastrointestinal and liver diseases. AB - Gene therapy consists of the transfer of genetic material to cells to achieve a therapeutic goal. In the field of gastroenterology and hepatology gene therapy has produced considerable expectation as a potential tool in the management of conditions that lack effective therapy including non-resectable neoplasms of the liver, pancreas and gastrointestinal tract, chronic viral hepatitis unresponsive to interferon therapy, liver cirrhosis, and inflammatory bowel disease. PMID- 12651883 TI - Fucose: biosynthesis and biological function in mammals. AB - Fucose is a deoxyhexose that is present in a wide variety of organisms. In mammals, fucose-containing glycans have important roles in blood transfusion reactions, selectin-mediated leukocyte-endothelial adhesion, host-microbe interactions, and numerous ontogenic events, including signaling events by the Notch receptor family. Alterations in the expression of fucosylated oligosaccharides have also been observed in several pathological processes, including cancer and atherosclerosis. Fucose deficiency is accompanied by a complex set of phenotypes both in humans with leukocyte adhesion deficiency type II (LAD II; also known as congenital disorder of glycosylation type IIc) and in a recently generated strain of mice with a conditional defect in fucosylated glycan expression. Fucosylated glycans are constructed by fucosyltransferases, which require the substrate GDP-fucose. Two pathways for the synthesis of GDP-fucose operate in mammalian cells, the GDP-mannose-dependent de novo pathway and the free fucose-dependent salvage pathway. In this review, we focus on the biological functions of mammalian fucosylated glycans and the biosynthetic processes leading to formation of the fucosylated glycan precursor GDP-fucose. PMID- 12651881 TI - Cholestasis. AB - In contrast with urine formation, bile flow is not dependent on hydrostatic forces, but driven by osmotic pressure of solutes secreted across the apical membrane of hepatocytes and bile duct epithelial cells. This secretory process is mediated by a set of primary active transporters that use ATP hydrolysis to pump solutes against the concentration gradient. The most important solutes in bile are bile salts, lipids, electrolytes, and organic anions. The direct consequence of the osmotic mechanism of bile formation is that impaired function of these pumps leads to impaired bile flow-that is, cholestasis. The function of these pumps is highlighted by a number of inherited cholestatic diseases, which are caused by mutations in these genes. Identification of the molecular defect in these diseases was not only important for diagnostic reasons but also emphasised that impaired transporter function has pathological consequences. Indeed, it is now becoming clear that impaired or downregulated transporter function is also involved in the pathogenesis of acquired cholestatic syndromes. PMID- 12651884 TI - Expression of UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase isoforms in murine tissues determined by real-time PCR: a new view of a large family. AB - The members of the UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase (ppGaNTase) family transfer GalNAc to serine and threonine sites and initiate mucin-type O-glycosylation. There are at least 13 functionally characterized family members in mammals. Explanations for the large size of this enzyme family have included functional redundancy, differences among isoforms in substrate specificity, and specific expression of individual isoforms in particular tissues or during certain developmental stages. To date no quantitative comparison of the levels of all ppGaNTase isoforms in any tissue of any species has been reported. We performed real-time polymerase chain reaction using the Taqman method to determine the expression of ppGaNTase isoforms in mouse tissues. Several tissues exhibited a common pattern in which isoforms T1 and T2 were the most strongly expressed, although the level of expression varied widely among tissues. In striking contrast to this general pattern, testis, sublingual gland, and colon exhibited distinctive profiles of isoform expression. Isoform T13 was expressed most strongly in brain, and one putative isoform was expressed only in testis. In mammary tissue the expression of several isoforms changed markedly during pregnancy and lactation. In summary these real-time PCR data indicate the contribution of each isoform to the overall ppGaNTase expression within each tissue and highlight the particular isoforms and tissues that will be the targets of future studies on the functions of the ppGaNTase family. PMID- 12651885 TI - An N-acetylglucosaminyltransferase of the Golgi apparatus of the yeast Saccharomyces cerevisiae that can modify N-linked glycans. AB - The yeast Saccharomyces cerevisiae is widely regarded as being only capable of producing N-linked glycans with high-mannose structures. To investigate the glycan structures made in different mutant strains, we made use of a reporter protein consisting of a version of hen egg lysozyme that contains a single site for N-linked glycosylation. Mass spectrometry analysis of the attached glycans revealed that a large proportion contained an unexpected extra mass corresponding to a single N-acetylhexosamine residue. In addition, the glycosylated lysozyme was recognized by an N-acetylglucosamine specific lectin. The genome of S. cerevisiae contains an uncharacterized open reading frame, YOR320c, that is related to a known N-acetylglucosaminyltransferase. Deletion of this ORF resulted in the disappearance of the extra mass on the N-linked glycans and loss of lectin binding. We show that the protein encoded by YOR320c (which we term Gnt1p) is localized to the Golgi apparatus and has GlcNAc-transferase activity in vitro. The physiological role of Gnt1p is unclear because mutants lacking the protein show no obvious growth or cell wall defects. Nonetheless, these results indicate that heterologous glycoproteins expressed in yeast can receive N-glycans with structures other than high mannose. In addition, they indicate that the lumen of the yeast Golgi contains UDP-GlcNAc, which may facilitate reconstitution of higher eukaryotic N-glycan processing. PMID- 12651886 TI - Mouse knockout solves endocrine puzzle and promotes new pituitary lineage model. PMID- 12651887 TI - Under arrest: cytostatic factor (CSF)-mediated metaphase arrest in vertebrate eggs. PMID- 12651888 TI - Human and mouse TPIT gene mutations cause early onset pituitary ACTH deficiency. AB - Tpit is a highly cell-restricted transcription factor that is required for expression of the pro-opiomelanocortin (POMC) gene and for terminal differentiation of the pituitary corticotroph lineage. Its exclusive expression in pituitary POMC-expressing cells has suggested that its mutation may cause isolated deficiency of pituitary adrenocorticotropin (ACTH). We now show that Tpit-deficient mice constitute a model of isolated ACTH deficiency (IAD) that is very similar to human IAD patients carrying TPIT gene mutations. Through genetic analysis of a panel of IAD patients, we show that TPIT gene mutations are associated at high frequency with early onset IAD, but not with juvenile forms of this deficiency. We identified seven different TPIT mutations, including nonsense, missense, point deletion, and a genomic deletion. This work defines congenital early onset IAD as a relatively homogeneous clinical entity caused by recessive transmission of loss-of-function mutations in the TPIT gene. PMID- 12651889 TI - Acetylation regulates subcellular localization of the Wnt signaling nuclear effector POP-1. AB - Lymphoid enhancer factor/T-cell factor (LEF/TCF) are transcription factors that mediate the Wnt signaling pathway, and have crucial roles during embryonic development in various organisms. Here we report that acetylation enhances nuclear retention of POP-1, the Caenorhabditis elegans LEF/TCF homolog, through increasing nuclear import and blocking nuclear export. We identify three lysines that are acetylated in vivo, and demonstrate their essential requirement for proper nuclear localization and biological activity of POP-1 during C. elegans embryogenesis. The conservation of these lysines among other LEF/TCF family members suggests that acetylation may be an important, evolutionarily conserved mechanism regulating subcellular distribution of LEF/TCF factors. PMID- 12651890 TI - G1 cyclin-dependent kinases are insufficient to reverse dE2F2-mediated repression. AB - Here we show that the cell cycle defects of dE2F1-depleted cells depend on the cooperative effects of dE2F2 and DACAPO (DAP), an inhibitor of Cyclin E/cyclin dependent kinase 2 (CycE/cdk2). The different properties of cells lacking dE2F1/dE2F2 and dE2F1/DAP lead to the surprising observation that dE2F2-mediated repression differs from retinoblastoma family protein 1 (RBF1) inhibition of dE2F1, and is resistant to both CycE/cdk2 and Cyclin D/cyclin-dependent kinase 4 (CycD/cdk4). This resistance occurs even though dE2F2/RBF1 complexes are disrupted by CycE/cdk2, and may explain why dE2F2 is so potent in the absence of de2f1. The implication of these results is that cells containing dE2F2 require dE2F1 to either prevent, or reverse, dE2F-mediated repression. PMID- 12651891 TI - Coordinated temporal and spatial control of motor neuron and serotonergic neuron generation from a common pool of CNS progenitors. AB - Neural progenitor cells often produce distinct types of neurons in a specific order, but the determinants that control the sequential generation of distinct neuronal subclasses in the vertebrate CNS remain poorly defined. We examined the sequential generation of visceral motor neurons and serotonergic neurons from a common pool of neural progenitors located in the ventral hindbrain. We found that the temporal specification of these neurons varies along the anterior-posterior axis of the hindbrain, and that the timing of their generation critically depends on the integrated activities of Nkx- and Hox-class homeodomain proteins. A primary function of these proteins is to coordinate the spatial and temporal activation of the homeodomain protein Phox2b, which in turn acts as a binary switch in the selection of motor neuron or serotonergic neuronal fate. These findings assign new roles for Nkx, Hox, and Phox2 proteins in the control of temporal neuronal fate determination, and link spatial and temporal patterning of CNS neuronal fates. PMID- 12651892 TI - Tpit determines alternate fates during pituitary cell differentiation. AB - The T-box transcription factor Tpit was identified as a cell-specific factor for expression of the pituitary proopiomelanocortin (POMC) gene. Expression of this factor is exclusively restricted to the pituitary POMC-expressing lineages, the corticotrophs and melanotrophs. We have now determined the role of this factor in pituitary cell differentiation. Tpit is a positive regulator for late POMC cell differentiation and POMC expression, but it is not essential for lineage commitment. The pituitary intermediate lobe normally contains only Tpit expressing melanotrophs. Inactivation of the Tpit gene results in almost complete loss of POMC-expressing cells in this tissue, which now has a large number of gonadotrophs and a few clusters of Pit-1-independent thyrotrophs. The role of Tpit as a negative regulator of gonadotroph differentiation was confirmed in transgenic gain-of-function experiments. One mechanism to account for the negative role of Tpit in differentiation may be trans-repression between Tpit and the gonadotroph-restricted factor SF1. These data suggest that antagonism between Tpit and SF1 may play a role in establishment of POMC and gonadotroph lineages and that these lineages may arise from common precursors. PMID- 12651893 TI - A model of repression: CTD analogs and PIE-1 inhibit transcriptional elongation by P-TEFb. AB - The positive transcription elongation factor b (P-TEFb) contains cyclin T1 (CycT1) and cyclin-dependent kinase 9 (Cdk9). For activating the expression of eukaryotic genes, the histidine-rich sequence in CycT1 binds the heptapeptide repeats in the C-terminal domain (CTD) of RNA polymerase II (RNAPII), whereupon Cdk9 phosphorylates the CTD. We found that alanine-substituted heptapeptide repeats that cannot be phosphorylated also bind CycT1. When placed near transcription units, these CTD analogs block effects of P-TEFb. Remarkably, the transcriptional repressor PIE-1 from Caenorhabditis elegans behaves analogously. It binds CycT1 via an alanine-containing heptapeptide repeat and inhibits transcriptional elongation. Thus, our findings reveal a new mechanism by which repressors inhibit eukaryotic transcription. PMID- 12651894 TI - Asynchronous replication timing of imprinted loci is independent of DNA methylation, but consistent with differential subnuclear localization. AB - Genomic imprinting in mammals marks the two parental alleles resulting in differential gene expression. Imprinted loci are characterized by distinct epigenetic modifications such as differential DNA methylation and asynchronous replication timing. To determine the role of DNA methylation in replication timing of imprinted loci, we analyzed replication timing in Dnmt1- and Dnmt3L deficient embryonic stem (ES) cells, which lack differential DNA methylation and imprinted gene expression. Asynchronous replication is maintained in these ES cells, indicating that asynchronous replication is parent-specific without the requirement for differential DNA methylation. Imprinting centers are required for regional control of imprinted gene expression. Analysis of replication fork movement and three-dimensional RNA and DNA fluorescent in situ hybridization (FISH) analysis of the Igf2-H19 locus in various cell types indicate that the Igf2-H19 imprinting center differentially regulates replication timing of nearby replicons and subnuclear localization. Based on these observations, we suggest a model in which cis elements containing nonmethylation imprints are responsible for the movement of parental imprinted loci to distinct nuclear compartments with different replication characteristics resulting in asynchronous replication timing. PMID- 12651895 TI - Recruitment of terminal protein to the ends of Streptomyces linear plasmids and chromosomes by a novel telomere-binding protein essential for linear DNA replication. AB - Bidirectional replication of Streptomyces linear plasmids and chromosomes from a central origin produces unpaired 3'-leading-strand overhangs at the telomeres of replication intermediates. Filling in of these overhangs leaves a terminal protein attached covalently to the 5' DNA ends of mature replicons. We report here the essential role of a novel 80-kD DNA-binding protein (telomere-associated protein, Tap) in this process. Biochemical studies, yeast two-hybrid analysis, and immunoprecipitation/immunodepletion experiments indicate that Tap binds tightly to specific sequences in 3' overhangs and also interacts with Tpg, bringing Tpg to telomere termini. Using DNA microarrays to analyze the chromosomes of tap mutant bacteria, we demonstrate that survivors of Tap ablation undergo telomere deletion, chromosome circularization, and amplification of subtelomeric DNA. Microarray-based chromosome mapping at single-ORF resolution revealed common endpoints for independent deletions, identified amplified chromosomal ORFs adjacent to these endpoints, and quantified the copy number of these ORFs. Sequence analysis confirmed chromosome circularization and revealed the insertion of adventitious DNA between joined chromosome ends. Our results show that Tap is required for linear DNA replication in Streptomyces and suggest that it functions to recruit and position Tpg at the telomeres of replication intermediates. They also identify hotspots for the telomeric deletions and subtelomeric DNA amplifications that accompany chromosome circularization. PMID- 12651896 TI - The yeast eIF3 subunits TIF32/a, NIP1/c, and eIF5 make critical connections with the 40S ribosome in vivo. AB - Initiation factor 3 (eIF3) forms a multifactor complex (MFC) with eIF1, eIF2, and eIF5 that stimulates Met-tRNA(i)(Met) binding to 40S ribosomes and promotes scanning or AUG recognition. We have previously characterized MFC subcomplexes produced in vivo from affinity-tagged eIF3 subunits lacking discrete binding domains for other MFC components. Here we asked whether these subcomplexes can bind to 40S ribosomes in vivo. We found that the N- and C-terminal domains of NIP1/eIF3c, the N- and C-terminal domains of TIF32/eIF3a, and eIF5 have critical functions in 40S binding, with eIF5 and the TIF32-CTD performing redundant functions. The TIF32-CTD interacted in vitro with helices 16-18 of domain I in 18S rRNA, and the TIF32-NTD and NIP1 interacted with 40S protein RPS0A. These results suggest that eIF3 binds to the solvent side of the 40S subunit in a way that provides access to the interface side for the two eIF3 segments (NIP1-NTD and TIF32-CTD) that interact with eIF1, eIF5, and the eIF2/GTP/Met-tRNA(i)(Met) ternary complex. PMID- 12651899 TI - The use of spermHALO-FISH to determine DAZ gene copy number. AB - The AZFc region of the human Y chromosome is frequently deleted in men with spermatogenic failure and contains many multicopy genes. The best-characterized gene family within this region is the Deleted in AZoospermia (DAZ) gene family, which is present in four nearly identical copies. Recent reports claim deletions of some but not all DAZ genes. The assays used in these studies, however, are unable to provide conclusive evidence on the number of DAZ genes. In this study we show that with the use of highly decondensed sperm nuclei with large DNA domains (spermHALO) it is possible to determine the number of DAZ genes accurately. Using this fluorescent in-situ hybridization (FISH) technique, which has both high resolution and high range, we show that in 10 normospermic men, in which PCR digest assays indicated a deletion of one or more DAZ genes, all four DAZ genes were present. Also we confirmed previous findings of a deletion of two DAZ genes in two men and identified a man with six DAZ genes. Our results indicate that spermHALO-FISH allows an accurate determination of DAZ gene copy number, while PCR digest assays do not. Therefore, confirmation of positive results from PCR digest assays with spermHALO-FISH is essential. Furthermore, the spermHALO-FISH technique should prove useful as a genetic mapping technique in other regions of the Y chromosome and similar repetitive regions throughout the genome. PMID- 12651898 TI - The molecular basis of cryptorchidism. AB - Cryptorchidism is the commonest malady to affect newborn male infants. Until recently, the molecular aetiology of this syndrome was unclear. Cryptorchidism may be part of a broader testicular dysgenesis syndrome, wherein a disturbance in steroid hormone metabolism, possibly through a perturbed hypothalamic-pituitary gonadal axis could be involved. Disturbance may be genetic, or extrinsic through endocrine disruptors. Recently, the role of insulin-like factor-3 (INSL3; alternatively called relaxin-like factor) has been highlighted through the cryptorchid phenotype of mice where genes for either INSL3 or its receptor have been ablated. INSL3 is produced by Leydig cells of the fetal testis and acts upon the gubernacular ligament to retain the gonad in the inguinal region, from which it later passes into the scrotum. INSL3 expression in fetal testis is inhibited by maternal exposure to estrogens. Although to date no mutations have been found in the human INSL3 gene responsible for cryptorchidism, one causative mutation in the INSL3 receptor (LGR8 or GREAT) has been reported. Studies on developmental transcription factors, such as Hoxa-10 in mice, suggest that other specific molecular cascades could also lead to a cryptorchid phenotype. Considering its frequency in newborn children, and the severity of the untreated condition (infertility and often testicular cancer) these new findings should generate new information on possible causes and treatments. PMID- 12651897 TI - Pdgfr-alpha mediates testis cord organization and fetal Leydig cell development in the XY gonad. AB - During testis development, the rapid morphological changes initiated by Sry require the coordinate integration of many signaling pathways. Based on the established role of the platelet-derived growth factor (PDGF) family of ligands and receptors in migration, proliferation, and differentiation of cells in various organ systems, we have investigated the role of PDGF in testis organogenesis. Analysis of expression patterns and characterization of the gonad phenotype in Pdgfr-alpha(-/-) embryos identified PDGFR-alpha as a critical mediator of signaling in the early testis at multiple steps of testis development. Pdgfr-alpha(-/-) XY gonads displayed disruptions in the organization of the vasculature and in the partitioning of interstitial and testis cord compartments. Closer examination revealed severe reductions in characteristic XY proliferation, mesonephric cell migration, and fetal Leydig cell differentiation. This work identifies PDGF signaling through the alpha receptor as an important event downstream of Sry in testis organogenesis and Leydig cell differentiation. PMID- 12651901 TI - Inhibin, activin, follistatin, activin receptors and beta-glycan gene expression in the placental tissue of patients with pre-eclampsia. AB - The objective of this study was to quantify the relative expression of inhibin alpha, inhibin/activin beta(A), beta(B), beta(C), follistatin, activin receptors and beta-glycan genes in placental tissue of term pre-eclamptic patients and controls to investigate if these genes are up-regulated in the placenta in pre eclampsia. Seven women with pre-eclampsia symptoms were matched with 10 normal pregnant controls for gestational age, maternal age, and parity. Total RNA was isolated from each sample. Complementary DNA samples produced by reverse transcription were used in the real time PCR to quantify the expression of inhibin alpha subunit, inhibin/activin beta(A), beta(B), beta(C) subunits, follistatin, ACTRIA, ACTRIB, ACTRIIA, ACTRIIB, beta-glycan and GAPDH genes. The ratio between the target and GAPDH expression was calculated to provide relative gene expression. Inhibin alpha:GAPDH and inhibin/activin beta(A): GAPDH ratios were significantly higher in placental tissue from women with pre-eclampsia (P = 0.04 and P = 0.01 respectively) compared with matched control placental gene expression. Placental samples from both groups expressed beta(B), beta(C), follistatin, activin receptors and beta-glycan genes. However, there was no significant difference in the relative expression of these genes between the groups. Increases in the placental expression of inhibin alpha and inhibin/activin beta(A) subunit genes could contribute to the rise in circulating levels of inhibin A and activin A in pre-eclampsia. The mechanism(s) involved in increased gene expression in pre-eclampsia is as yet unclear. PMID- 12651900 TI - Hormonal and embryonic regulation of chemokine receptors CXCR1, CXCR4, CCR5 and CCR2B in the human endometrium and the human blastocyst. AB - Chemokines are implicated in the implantation process. The aim of this study was to investigate mRNA expression and protein levels of chemokine receptors CXCR1, CXCR4, CCR5 and CCR2B in human endometrium throughout the menstrual cycle, during HRT and in the human blastocyst. The regulation of chemokine receptors in the endometrial epithelium was also studied using an in-vitro model for the apposition phase of human implantation. We found up-regulation of endometrial CXCR1 mRNA (419-fold increase), CCR5 mRNA (612-fold increase) and CCR2B mRNA (657 fold-increase) during the luteal phase peaking in the pre-menstrual endometrium. CXCR4 mRNA levels presented a specific although modest (18-fold increase) up regulation during the implantation window. These findings were corroborated at the protein level in natural and HRT cycles. Immunoreactive CCR5 and CCR2B receptors were detected in human blastocysts whereas CXCR4 and CXCR1 were not present. Chemokine receptors in cultured endometrial epithelial cells showed an up-regulation and polarization of CXCR1, CXCR4 and CCR5 receptors when a human blastocyst was present. The specific distribution and regulation of chemokine receptors in the endometrial epithelium and the human blastocyst suggest a possible implication of these receptors in the apposition and adhesion phases of human implantation. PMID- 12651902 TI - Matrix metalloproteinase-28 transcript and protein are expressed in rhesus monkey placenta during early pregnancy. AB - The role of matrix metalloproteinases (MMP), especially newly described MMP, in trophoblast invasion during human embryo implantation is poorly understood. In this report, using a model of early pregnancy in the rhesus monkey, we have examined the expression and localization of the most recently identified MMP, MMP 28/epilysin, transcript and protein in macaque uterine samples on days 12, 18 and 26 of pregnancy. MMP-28 mRNA expression was shown by in-situ hybridization after day 12 of pregnancy, and both the syncytial and the cytotrophoblastic cell layers of placental villi, the cytotrophoblast cells of the trophoblastic column, and the extravillous trophoblast cells of trophoblastic shell were primary producers of MMP-28 transcript. Expression of MMP-28 mRNA was undetectable in the endovascular trophoblast cells, decidual cells, luminal and glandular epithelium, arterioles, and myometrium. RT-PCR analysis amplified a fragment of 258 nucleotides from rhesus monkey uterine samples containing implantation sites on days 18 and 26. The cDNA fragment, following sequencing, was confirmed to be part of the haemopexin-like domain of MMP-28. It has 95% identity with the corresponding region of human MMP-28 gene. Immunohistochemical analysis further demonstrated that the localization of MMP-28 protein was similar to that of its mRNA. The restricted distribution pattern of this novel MMP in the villous and extravillous trophoblasts during rhesus monkey early pregnancy suggests a potential role in trophoblast invasion associated with embryo implantation. PMID- 12651903 TI - The effects of labour and of interleukin 1 beta upon the expression of nuclear factor kappa B related proteins in human amnion. AB - Human labour is associated with persistently increased nuclear factor kappa B (NF kB) activity in amnion. In this study we have shown that this involves only the p65 and p50 NF-kB subunits and is associated with an increase in the expression of p65 (P < 0.05), and of the NF-kB binding proteins IkBa, IkBb-1 and IkBb-2 (P < 0.05). Interleukin-1b stimulation leads to rapid degradation and resynthesis of IkBa within 2 h, and a decrease in IkBb-1 without a return to full expression by 2 h, but has little effect upon IkBb-2. IkBb-2 was found in both the cytosolic and nuclear protein fractions. These findings demonstrate that persistently increased NF-kB activity in amnion occurs despite increased expression of the inhibitory IkBa protein and is not mediated by persistant I-kappa kinase activity or inhibition of IkBa synthesis. The increased expression and nuclear localization of IkBb-2 suggests that its function may be to protect NF-kB from inactivation by IkBa and to maintain NF-kB-mediated gene transcription. PMID- 12651904 TI - Accelerated germ cell apoptosis in sex chromosome aneuploid fetal human gonads. AB - The purpose of the study was to examine the occurrence of programmed cell death (apoptosis) in normal and chromosomally aneuploid testis and ovaries during the second trimester of human development. Such information may be useful in understanding normal and abnormal germ cell development and disorders associated with infertility in adult life. Apoptosis was studied by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) analysis in human fetal ovaries (n = 16) and testis (n = 14) between 9 and 23 weeks of development, in ovaries of four Turner's syndrome fetuses (45X) and in the gonad of an XO/XY fetus. In normal fetal testis, a small proportion of germ cells, Sertoli cells and Leydig cells undergo apoptosis. In normal fetal ovaries, some developing oocytes and granulosa cells were detected as TUNEL positive. Semiquantitative analysis of fetal ovaries revealed that approximately 3-7% of oocytes were apoptotic. In abnormal fetal testis (XO/XY genotype). TUNEL analysis revealed that only germ cells not enclosed in seminiferous tubules undergo apoptosis. TUNEL analysis of the Turner's syndrome (45X) ovaries studied at 15 and 20 weeks of development revealed massive apoptosis of the oocytes. Nearly 50 70% of the oocytes were TUNEL positive in these ovaries. These results suggest that germ cell apoptosis is a common event occurring during development of human gonads. Chromosomal defects by some means accelerates apoptosis that probably leads to gonadal dysgenesis later in life. PMID- 12651905 TI - Characterization of first trimester fetal erythroblasts for non-invasive prenatal diagnosis. AB - Isolating fetal erythroblasts from first trimester maternal blood offers a promising non-invasive alternative for prenatal diagnosis. The aim of this study was to characterize the biological properties of first trimester primitive erythroblasts to facilitate their enrichment from first trimester maternal blood. Primitive erythroblasts were the predominant cell type until 12 weeks gestation, after which time their numbers declined steeply; 100% were epsilon-globin positive versus <0.06% definitive erythroblasts. Buoyant densities of first trimester fetal erythroblasts ranged from 1.077 to 1.130 g/ml, and optimal recoveries were obtained with Percoll 1118. Although primitive erythroblasts carried a negative surface charge and were resistant to NH(4)Cl lysis, these properties had only a limited role in fetal cell enrichment. Immunophenotyping showed that primitive, like definitive, erythroblasts were GPA+, CD47+, CD45- and CD35-, whereas CD71 expression was weak/undetectable on primitive erythroblasts but strongly positive on 100% of definitive erythroblasts; primitive erythroblasts were also CD36- whereas definitive erythroblasts were CD36+. We therefore used CD45/GPA selection of Percoll 1118-separated cells to demonstrate successful enrichment of male epsilon-globin-positive fetal erythroblasts from model mixtures, and as proof of principle from some first trimester maternal blood samples. Fetal cell enrichment protocols based on first trimester epsilon globin-positive primitive erythroblasts may allow reliable enrichment of fetal cells from maternal blood for early non-invasive prenatal diagnosis of genetic disorders. PMID- 12651906 TI - Differential role of proline-rich tyrosine kinase 2 and focal adhesion kinase in determining glioblastoma migration and proliferation. AB - The propensity of malignant gliomas to invade surrounding brain tissue contributes to poor clinical outcome. Integrin-mediated adhesion to extracellular matrix regulates the migration and proliferation of many cell types, but its role in glioma progression is undefined. We investigated the role of the cytoplasmic tyrosine kinases FAK and Pyk2, potential integrin effectors, in the phenotypic determination of four different human glioblastoma cell lines. While FAK expression was similar between the four cell lines, increased FAK activity correlated with high proliferation and low migratory rates. In contrast, Pyk2 activity was significantly increased in migratory cell lines and depressed in proliferative cell lines. Overexpression of Pyk2 stimulated migration, whereas FAK overexpression inhibited cell migration and stimulated cellular proliferation. These data suggest that FAK and Pyk2 function as important signaling effectors in gliomas and indicate that their differential regulation may be determining factors in the temporal development of proliferative or migrational phenotypes. PMID- 12651907 TI - Elevated membrane-type matrix metalloproteinases in gliomas revealed by profiling proteases and inhibitors in human cancer cells. AB - Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) regulate proteolysis of the extracellular matrix and other extracellular proteins, including growth factors and their receptors. The aberrant expression of these genes is common in most cancers. We profiled the RNA levels of every human MMP and TIMP in a variety of cell types (fibroblast, endothelial, hematopoietic, carcinoma, melanoma, and glioma) using quantitative PCR, with the aim of identifying novel expression patterns. Almost all members of the membrane type (MT-) MMP and TIMP families were elevated in glioma lines compared to carcinomas. In clinical glioma specimens, there were positive correlations between glioma grade and RNA levels of MT-1, MT-2, and MT-6 MMP, TIMP-1 and TIMP 2, and for several growth factors and receptors. These findings suggest that advanced malignant gliomas have elevated levels of membrane-associated MMPs and TIMPs, which may potentially regulate vascularization and invasion. Concurrent elevation of signaling molecules suggests potential bidirectional relationships that enhance tumor aggressiveness. PMID- 12651908 TI - Identification of a global gene expression signature of B-chronic lymphocytic leukemia. AB - B-chronic lymphocytic leukemia (B-CLL) is an adult-onset leukemia characterized by significant accumulation of apoptosis-resistant monoclonal B lymphocytes. In this study, we performed gene expression profiling on B cells obtained from 10 healthy age-matched individuals and CLL B cells from 38 B-CLL patients to identify key genetic differences between CLL and normal B cells. In addition, we leveraged recent independent studies to assess the reproducibility of our molecular B-CLL signature. We used a novel combination of several methods of data analysis including our own software and identified 70 previously unreported genes that differentiate leukemic cells from normal B cells, as well as confirmed recently reported B-CLL specific expression levels of an additional 10 genes. Importantly, many of these genes have previously been linked with other cancers, thus lending further support to their importance as candidate genes leading to B CLL pathogenesis. We have also validated a subset of these genes using independent methodologies. Moreover, we show that our genes can be used to create a diagnostics signature that performs with perfect sensitivity and specificity in an independent cohort of 21 B-CLL and 20 normal subjects, thus strongly validating the informative nature of our set of genes. Finally, we identified a group of 31 genes that distinguish between low (Rai stage 0) and high (Rai stage 4) risk patients, suggesting that there may also be a gene expression signature that associates with disease progression. PMID- 12651909 TI - Molecular markers in ductal carcinoma in situ of the breast. AB - Gene expression patterns in ductal carcinoma in situ (DCIS), and in invasive, and metastatic breast tumors were determined using serial analysis of gene expression (SAGE). We used mRNA in situ hybridization to examine gene expression at the cellular level and immunohistochemistry on tissue microarrays to determine association between gene expression patterns and histopathologic characteristics of the tumors. We found that that the most dramatic transcriptome change occurs at the normal to DCIS transition, while there is no clear universal "in situ" or "invasive" tumor molecular signature. From the 16,430 transcripts analyzed, we identified only 5 and 11 that were preferentially up-regulated in DCIS and invasive tumors, respectively. The majority of invasive cancer specific SAGE tags correspond to novel genes. The genes we identified may define biologically and clinically meaningful subgroups of DCIS with a high risk of progression to invasive disease. PMID- 12651910 TI - The serum-inducible protein kinase Snk is a G1 phase polo-like kinase that is inhibited by the calcium- and integrin-binding protein CIB. AB - Identified as an immediate-early transcript, the serum-inducible kinase Snk bears sequence homology with the polo-like kinases. Endogenous Snk was detected in early G(1) in NIH 3T3 cells, and nascent Snk showed a half-life of about 15 min. The kinase activity of endogenous Snk was detected in G(1). Substitution of Thr 236 with a glutamate residue increased Snk kinase activity by about 10-fold, whereas substitution of Lys-108 abolished its kinase activity. Disrupting the polo-box did not significantly change Snk kinase activity. A GFP-C-Snk fusion protein showed polo-box-dependent localization to the microtubule organizing center, and ectopic expression of Snk in COS-7 cells induced changes in cell morphology, depending on Snk kinase activity and the polo-box. The capacity of Snk to induce morphological changes was inhibited by the calcium- and integrin binding protein CIB. CIB co-immunoprecipitated with Snk and inhibited the kinase activity of Snk, suggesting that CIB is a negative regulator for Snk kinase activity. PMID- 12651911 TI - Androgen receptor activity at the prostate specific antigen locus: steroidal and non-steroidal mechanisms. AB - Ligand-activated androgen receptors (ARs) occupy target genes and recruit histone modifiers that influence transcriptional competency. In LNCaP prostate cancer cells, the natural ligand 5alpha-dihydrotestosterone (DHT) activates transiently transfected AR-responsive promoter constructs; concurrent treatment with the protein kinase A activator forskolin enhanced AR stimulation induced by DHT. Additional treatment with the cytokine IL-6, purportedly an AR activator, markedly inhibited receptor activity. To assess AR activity on natural chromatin integrated promoters/enhancers, we determined AR occupancy of the endogenous prostate specific antigen (PSA) promoter/enhancer as well as PSA expression in LNCaP cells treated with DHT; AR occupancy of the PSA enhancer was rapid (within 1 h of stimulation), robust (10-fold over background), and sustained (8-16 h). In contrast, AR occupancy of the PSA promoter was only increased by 2-fold. Histone H3 acetylation at both the enhancer and promoter was evident 1-2 h after DHT treatment. Detectable pre- and mature PSA mRNA levels appeared after 1 and 6 h treatment, respectively. Substantial qualitative and quantitative differences in PSA expression and AR occupancy of the PSA enhancer were observed when DHT induced and ligand-independent activations of the AR were compared; forskolin stimulated PSA mRNA and protein expression, whereas IL-6 inhibited both DHT- and forskolin-stimulated expression. IL-6 did not diminish DHT-dependent AR occupancy of the PSA enhancer but inhibited CBP/p300 recruitment, histone H3 acetylation, and cell proliferation. These findings provide a contextual framework for interpreting the contribution of non-steroidal activation of the AR to signaling in vivo, and have implications for prostate cancer cell growth. PMID- 12651912 TI - Met receptor overexpression and oncogenic Ki-ras mutation cooperate to enhance tumorigenicity of colon cancer cells in vivo. AB - We have investigated the influence of Ki-ras oncogene on Met/hepatocyte growth factor (HGF) receptor signaling in human carcinoma cells. The model system used in these studies included the DLD-1 colon cancer cell line with a mutated Ki-ras allele, and the DKO-4 cell line generated from DLD-1, with its mutant Ki-ras allele inactivated by targeted disruption. These cell lines were transduced with cDNAs of either active Met receptor or dominant negative Met receptor. As compared to the DLD-1 cells, constitutive overexpression of Met receptor in this cell line (DLD-1-Met) resulted in increased tumorigenicity in SCID mice. In contrast, overexpression of Met in DKO-4 cells (DKO-4-Met) that have lost oncogenic Ras activity demonstrated suppressed tumorigenicity with respect to the parent DKO-4 cell line. Tumors formed by the DLD-1-Met cells showed increased levels of mitogen-activated protein kinase (MAPK) and lower levels of apoptosis compared to the DKO-4-Met tumors. Overexpression of the dominant negative Met receptor cDNA decreased the Met phosphorylation levels in both DLD-1 and DKO-4 cells, but only suppressed tumorigenicity in the DKO-4 cell line. In vitro, HGF stimulation of DLD-1 cells resulted in a prolonged duration of MAPK activation, while DKO-4 cells exhibited a rapid attenuation of MAPK phosphorylation. The results suggest that Ki-ras mutations and HGF signaling cooperate to enhance tumor growth by increased duration of MAPK activation and decreased apoptosis in human carcinoma cells. PMID- 12651913 TI - The absence of SIR2alpha protein has no effect on global gene silencing in mouse embryonic stem cells. AB - The yeast sir2 gene plays a central role in mediating gene silencing and DNA repair in this organism. The mouse sir2alpha gene is closely related to its yeast homologue and encodes a nuclear protein expressed at particularly high levels in embryonic stem (ES) cells. We used homologous recombination to create ES cells null for sir2alpha and found that these cells did not have elevated levels of acetylated histones and did not ectopically express silent genes. Unlike yeast sir2 mutants, our sir2alpha null ES cells had normal sensitivity to insults such as ionizing radiation and heat shock, and they were able to silence invading retroviruses normally. These sir2alpha null cells were able to differentiate in culture normally. Our results failed to provide evidence that the mammalian SIR2alpha protein plays a role in gene silencing and suggest that the physiological substrate(s) for the SIR2alpha deacetylase may be nuclear proteins other than histones. PMID- 12651914 TI - Skeletal muscle fat and carbohydrate metabolism during recovery from glycogen depleting exercise in humans. AB - The primary aim of the present study was to determine whether intramuscular triacylglycerol (IMTG) utilization contributed significantly to the increase in lipid oxidation during recovery from exercise, as determined from the muscle biopsy technique. In addition, we also examined the regulation of pyruvate dehydrogenase (PDHa) and changes in muscle acetyl units during an 18 h recovery period after glycogen-depleting exercise. Eight endurance-trained males completed an exhaustive bout of exercise (approximately 90 min) on a cycle ergometer followed by ingestion of carbohydrate (CHO)-rich meals (64-70 % of energy from carbohydrate) at 1, 4 and 7 h of recovery. Duplicate muscle biopsies were obtained at exhaustion, and 3, 6 and 18 h of recovery. Despite the large intake of CHO during recovery (491 +/- 28 g or 6.8 +/- 0.3 g kg-1), respiratory exchange ratio values of 0.77 to 0.84 indicated a greater reliance on lipid as an oxidative fuel. However, there was no net IMTG utilization during recovery. IMTG content at exhaustion was 23.5 +/- 3.5 mmol (kg dry wt)-1, and remained constant at 24.6 +/- 2.6, 25.7 +/- 2.8 and 28.4 +/- 3.0 mmol (kg dry wt)-1 after 3, 6 and 18 h of recovery. Muscle glycogen increased significantly from 37 +/- 11 mmol (kg dry wt)-1 at exhaustion, to 165 +/- 13, 250 +/- 18, and 424 +/- 22 mmol (kg dry wt)-1 at 3, 6 and 18 h of recovery, respectively. PDHa was reduced at 6 and 18 h when compared to exhaustion, but did not change during the recovery period. Acetyl-CoA, acetylcarnitine and pyruvate contents declined significantly after 3 h of recovery compared to exhaustion, and thereafter remained unchanged. We conclude that IMTG has a negligible role in contributing to the enhanced fat oxidation during recovery from exhaustive exercise. Despite the elevation of glucose and insulin following high-CHO meals during recovery, CHO oxidation and PDH activation were decreased, supporting the hypothesis that glycogen resynthesis is of high metabolic priority. Plasma fatty acids, very low density lipoprotein triacylglycerols, as well as intramuscular acetylcarnitine stores are likely to be important fuel sources for aerobic energy, particularly during the first few hours of recovery. PMID- 12651915 TI - Angiotensin II-induced modulation of endothelium-dependent relaxation in rabbit mesenteric resistance arteries. AB - The role of local endogenous angiotensin II (Ang II) in endothelial function in resistance arteries was investigated using rabbit mesenteric resistance arteries. First, the presence of immunoreactive Ang II together with Ang II type-1 receptor (AT1R) and angiotensin converting enzyme (ACE) was confirmed in these arteries. In endothelium-intact strips, the AT1R-blocker olmesartan (1 microM) and the ACE inhibitor temocaprilat (1 microM) each enhanced the ACh (0.03 microM)-induced relaxation during the contraction induced by noradrenaline (NA, 10 microM). Similar effects were obtained using CV-11974 (another AT1R blocker) and enalaprilat (another ACE inhibitor). The nitric-oxide-synthase inhibitor NG-nitro L-arginine (L-NNA) abolished the above effect of olmesartan. In endothelium denuded strips, olmesartan enhanced the relaxation induced by the NO donor NOC-7 (10 nM). Olmesartan had no effect on cGMP production (1) in endothelium-intact strips (in the absence or presence of ACh) or (2) in endothelium-denuded strips (in the absence or presence of NOC-7). In beta-escin-skinned strips, 8 bromoguanosine 3',5' cyclic monophosphate (8-Br-cGMP, 0.01-1 microM) concentration dependently inhibited the contractions induced (a) by 0.3 microM Ca2+ in the presence of NA+GTP and (b) by 0.2 microM Ca2++GTPgammaS. Olmesartan significantly enhanced, while Ang II (0.1 nM) significantly inhibited, the 8-Br cGMP-induced relaxation. We propose the novel hypothesis that in these arteries, Ang II localized within smooth muscle cells activates AT1Rs and inhibits ACh induced, endothelium-dependent relaxation at least partly by inhibiting the action of cGMP on these cells. PMID- 12651917 TI - ATP and heat production in human skeletal muscle during dynamic exercise: higher efficiency of anaerobic than aerobic ATP resynthesis. AB - The aim of the present study was to simultaneously examine skeletal muscle heat production and ATP turnover in humans during dynamic exercise with marked differences in aerobic metabolism. This was done to test the hypothesis that efficiency is higher in anaerobic than aerobic ATP resynthesis. Six healthy male subjects performed 90 s of low intensity knee-extensor exercise with (OCC) and without thigh occlusion (CON-LI) as well as 90 s of high intensity exercise (CON HI) that continued from the CON-LI bout. Muscle heat production was determined by continuous measurements of muscle heat accumulation and heat release to the blood. Muscle ATP production was quantified by repeated measurements of thigh oxygen uptake as well as blood and muscle metabolite changes. All temperatures of the thigh were equalized to approximately 37 degrees C prior to exercise by a water-perfused heating cuff. Oxygen uptake accounted for 80 +/- 2 and 59 +/- 4 %, respectively, of the total ATP resynthesis in CON-LI and CON-HI, whereas it was negligible in OCC. The rise in muscle temperature was lower (P < 0.05) in OCC than CON-LI (0.32 +/- 0.04 vs. 0.37 +/- 0.03 degrees C). The mean rate of heat production was also lower (P < 0.05) in OCC than CON-LI (36 +/- 4 vs. 57 +/- 4 J s-1). Mechanical efficiency was 52 +/- 4 % after 15 s of OCC and remained constant, whereas it decreased (P < 0.05) from 56 +/- 5 to 32 +/- 3 % during CON LI. During CON-HI, mechanical efficiency transiently increased (P < 0.05) to 47 +/- 4 %, after which it decreased (P < 0.05) to 36 +/- 3 % at the end of CON-HI. Assuming a fully coupled mitochondrial respiration, the ATP turnover per unit of work was calculated to be unaltered during OCC (approximately 20 mmol ATP kJ-1), whereas it increased (P < 0.05) from 21 +/- 4 to 29 +/- 3 mmol ATP kJ-1 during CON-LI and further (P < 0.05) to 37 +/- 3 mmol ATP kJ-1 during CON-HI. The present data confirm the hypothesis that heat loss is lower in anaerobic ATP resynthesis than in oxidative phosphorylation and can in part explain the finding that efficiency declines markedly during dynamic exercise. In addition, the rate of ATP turnover apparently increases during constant load low intensity exercise. Alternatively, mitochondrial efficiency is lowered as exercise progresses, since ATP turnover was unaltered during the ischaemic exercise bout. PMID- 12651916 TI - Chick RGS2L demonstrates concentration-dependent selectivity for pertussis toxin sensitive and -insensitive pathways that inhibit L-type Ca2+ channels. AB - In neuronal cells, the influx of Ca2+ ions through voltage-dependent L-type calcium (L) channels couples excitation to multiple cellular functions. In addition to voltage, several neurotransmitters, hormones and cytokines regulate L channel gating via binding to G-protein-coupled receptors. Intracellular molecules that modify G-protein activity - such as regulator of G-protein signalling (RGS) proteins - are therefore potential candidates for regulating Ca2+ influx through L channels. Here we show that a novel RGS2 splice variant from chick dorsal root ganglion (DRG) neurons, RGS2L, reduces bradykinin (BK) mediated inhibition of neuronal L channels and accelerates recovery from inhibition. Chick RGS2 reduces the inhibition mediated by both the pertussis toxin (PTX)-sensitive (Gi/o-coupled) and the PTX-insensitive (presumably Gq/11 coupled) pathways. However, we demonstrate for the first time in a living cell that the extent of coupling to each pathway varies with RGS2L concentration. A low concentration of recombinant chick RGS2L (10 nM) preferentially reduces the inhibition mediated by the PTX-insensitive pathway, whereas a 100-fold higher concentration attenuates both PTX-sensitive- and PTX-insensitive-mediated components equally. Our data suggest that factors promoting RGS2L gene induction may regulate Ca2+ influx through L channels by recruiting low-affinity interactions with Gi/o that are absent at basal RGS2L levels. PMID- 12651918 TI - Nitric oxide is not permissive for cutaneous active vasodilatation in humans. AB - The precise role of nitric oxide (NO) in cutaneous active vasodilatation in humans is unknown. We tested the hypothesis that NO is necessary to permit the action of an unknown vasodilator. Specifically, we investigated whether a low dose infusion of exogenous NO, in the form of sodium nitroprusside (SNP), would fully restore vasodilatation in an area of skin in which endogenous NO was inhibited during hyperthermia. This finding would suggest a 'permissive' role for NO in active vasodilatation. Eight subjects were instrumented with three microdialysis fibres in forearm skin. Sites were randomly assigned to (1) Site A: control site; (2) Site B: NO synthase (NOS) inhibition during established hyperthermia; or (3) Site C: NOS inhibition throughout the protocol. Red blood cell flux was measured using laser-Doppler flowmetry (LDF) and cutaneous vascular conductance (CVC; LDF/mean arterial pressure) was normalized to maximal vasodilatation at each site. In Site B, NG-nitro-L-arginine methyl ester (L-NAME) infusion during hyperthermia reduced CVC by approximately 32 % (65 +/- 4 % CVCmax vs. 45 +/- 4 % CVCmax; P < 0.05). Vasodilatation was not restored to pre-NOS inhibition values in this site following low-dose SNP infusion (55 +/- 4 % CVCmax vs. 65 +/- 4 % CVCmax; P < 0.05). CVC remained significantly lower than the control site with low-dose SNP infusion in Site C (P < 0.05). The rise in CVC with low-dose SNP (deltaCVC) was significantly greater in Site B and Site C during hyperthermia compared to normothermia (P < 0.05). No difference in deltaCVC was observed between hyperthermia and normothermia in the control site (Site A). Thus, NO does not act permissively in cutaneous active vasodilatation in humans but may directly mediate vasodilatation and enhance the effect of an unknown active vasodilator. PMID- 12651919 TI - A-, T-, and H-type currents shape intrinsic firing of developing rat abducens motoneurons. AB - During postnatal development, profound changes take place in the excitability of nerve cells, including modification in the distribution and properties of receptor-operated channels and changes in the density and nature of voltage-gated channels. We studied here the firing properties of abducens motoneurons (aMns) in transverse brainstem slices from postnatal day (P) 1-13 rats. Recordings were made from aMNs in the whole-cell configuration of the patch-clamp technique. Two main types of aMn could be distinguished according to their firing profile during prolonged depolarizations. Both types were identified as aMns by their fluorescence following retrograde labelling with the lipophilic carbocyanine DiI in the rectus lateralis muscle. The first type (BaMns) exhibited a burst of action potentials (APs) followed by an adaptation of discharge and were encountered in approximately 70 % of aMns. Their discharge profile resembled that of adult aMns and was encountered in all aMns after P9. BaMns exhibited a hyperpolarization-induced rebound potential that was blocked by low concentrations of Ni2+ or by Ca2+-free external solution. This current had the properties of the T-type current. Action potentials of BaMns showed a complex afterhyperpolarization (AHP). An inward rectification was evidenced following hyperpolarization and was blocked by external application of caesium or ZD7288, indicating the presence of the hyperpolarization-activated cationic current (IH). Blocking the IH current almost doubled the input resistance of BaMns. The second class of aMns (DaMns) displayed a delayed excitation that was mediated by A-type K+ currents and was observed only between P4 and P9. DaMns exhibited immature characteristics: an action potential with a simple AHP, a linear current-voltage relation and a large input resistance. The number of aMns remained unchanged when both types were present (P5-P6) and later in development when only BaMns were encountered (P19), suggesting that DaMns mature into BaMns during postnatal development. We conclude that aMns display profound reorganization in their intrinsic excitability during postnatal development. PMID- 12651920 TI - Kappa-opioid receptor-mediated enhancement of the hyperpolarization-activated current (I(h)) through mobilization of intracellular calcium in rat nucleus raphe magnus. AB - The hyperpolarization-activated current (Ih) is important in the control of resting membrane potential, in the regulation of network firing pattern and in the modulation of presynaptic transmitter release in central neurons. Recent studies on native and cloned Ih channels have demonstrated that the Ih channel is commonly modulated by cAMP through a positive shift in its voltage dependence without a change in its maximum current. The present study demonstrates that activation of kappa-opioid receptors enhances Ih by increasing its maximum current in brainstem neurons in the nucleus raphe magnus. Agents that interfere with the release of intracellular calcium from calcium stores altered the maximum Ih and significantly attenuated the kappa-receptor-mediated enhancement of Ih. These results suggest that kappa-opioid receptors enhance the maximum Ih by mobilizing intracellular calcium from calcium stores. This provides a physiological function for kappa-receptor-stimulated calcium release and may suggest another Ih-regulating mechanism by intracellular calcium in central neurons. PMID- 12651921 TI - Interstitial exclusion of albumin in rabbit lung during development of pulmonary oedema. AB - The modifications of the macromolecular sieving properties of the pulmonary extracellular tissue matrix were studied in adult anaesthetized rabbits (n = 10) exposed to increased tissue hydration. Exclusion of albumin from the perivascular pulmonary interstitial space was determined by using the continuous infusion method coupled with direct sampling of interstitial fluid performed through the wick technique. The rabbits underwent an intravenous infusion of saline amounting to 10 (n = 5) or 20 % (n = 5) body weight. Extracellular albumin distribution volume was derived from the steady state tissue concentration of radioactive rabbit serum albumin (125I-RSA). Pulmonary extracellular and intravascular fluid volumes (Vx and Vv, respectively) were measured as distribution volumes of 51Cr EDTA and 131I-RSA, respectively, and interstitial fluid tracer concentrations were determined in interstitial fluid collected through implanted wicks. At the highest degree of hydration, interstitial fluid volume (Vi = Vx - Vv) and extravascular albumin distribution volume (Va,w) significantly increased by 38.5 and 240.2 %, respectively, compared to control. Albumin-excluded volume (Ve,a = Vi - Va,w) was 398.9 +/- 17 microl (g dry tissue weight)-1; the albumin-excluded volume fraction (Fe,a = Ve,a/Vi) was 0.23 +/- 0.01, 33.2 % of the control value. Data indicate that, at variance with what is observed in tissues like skin and muscle, pulmonary Fe,a is highly sensitive to tissue fluid content. PMID- 12651922 TI - GTP-induced tetrodotoxin-resistant Na+ current regulates excitability in mouse and rat small diameter sensory neurones. AB - Peripheral pain thresholds are regulated by the actions of inflammatory mediators. Some act through G-protein-coupled receptors on voltage-gated sodium channels. We have found that a low-threshold, persistent tetrodotoxin-resistant Na+ current, attributed to NaV1.9, is upregulated by GTP and its non-hydrolysable analogue GTP-gamma-S, but not by GDP. Inclusion of GTP-gamma-S (500 microM) in the internal solution led to an increase in maximal current amplitude of > 300 % within 5 min. In current clamp, upregulation of persistent current was associated with a more negative threshold for action potential induction (by 15-16 mV) assessed from a holding potential of -90 mV. This was not seen in neurones without the low-threshold current or with internal GDP (P < 0.001). In addition, persistent current upregulation depolarized neurones. At -60 mV, internal GTP gamma-S led to the generation of spontaneous activity in initially silent neurones only when persistent current was upregulated. These findings suggest that regulation of the persistent current has important consequences for nociceptor excitability. PMID- 12651924 TI - Sympathetic vasodilatation in human skin. PMID- 12651925 TI - Smallpox: the main site of transmission is the oropharynx. PMID- 12651926 TI - Dental caries seen from the pulpal side: a non-traditional approach. PMID- 12651927 TI - Dissociation states of collagen functional groups and their effects on the priming efficacy of HEMA bonded to collagen. AB - Applying 2-hydroxyethylmethacrylate (HEMA) solution to etched dentin enhances the bonding of resin to dentin. However, the principal adhesion mechanisms have not yet been identified. In this study, we examined the dissociation states of the collagen functional groups of the side-chain amino acid residues and their effects on the bond strength of resin to etched dentin primed by the HEMA solution. The bond strength was strongly dependent upon the dissociation state of the collagen functional groups. Inhibiting the dissociation of the carboxylic acid or the amine of a collagen functional group resulted in increased bond strength of resin to collagen. By understanding the significance of inhibiting the dissociation state, we can better design and develop more effective and efficient primer and bonding agents. PMID- 12651923 TI - Acute regulation of the SLC26A3 congenital chloride diarrhoea anion exchanger (DRA) expressed in Xenopus oocytes. AB - Mutations in the human SLC26A3 gene, also known as down-regulated in adenoma (hDRA), cause autosomal recessive congenital chloride-losing diarrhoea (CLD). hDRA expressed in Xenopus oocytes mediated bidirectional Cl--Cl- and Cl--HCO3- exchange. In contrast, transport of oxalate was low, and transport of sulfate and of butyrate was undetectable. Two CLD missense disease mutants of hDRA were nonfunctional in oocytes. Truncation of up to 44 C-terminal amino acids from the putatively cytoplasmic C-terminal hydrophilic domain left transport function unimpaired, but deletion of the adjacent STAS (sulfate transporter anti-sigma factor antagonist) domain abolished function. hDRA-mediated Cl- transport was insensitive to changing extracellular pH, but was inhibited by intracellular acidification and activated by NH4+ at acidifying concentrations. These regulatory responses did not require the presence of either hDRA's N-terminal cytoplasmic tail or its 44 C-terminal amino acids, but they did require more proximate residues of the C-terminal cytoplasmic domain. Although only weakly sensitive to inhibition by stilbenes, hDRA was inhibited with two orders of magnitude greater potency by the anti-inflammatory drugs niflumate and tenidap. cAMP-insensitive Cl--HCO3- exchange mediated by hDRA gained modest cAMP sensitivity when co-expressed with cystic fibrosis transmembrane conductance regulator (CFTR). Despite the absence of hDRA transcripts in human cell lines derived from CFTR patients, DRA mRNA was present at wild-type levels in proximal colon and nearly so in the distal ileum of CFTR(-/-) mice. Thus, pharmacological modulation of DRA might be a useful adjunct treatment of cystic fibrosis. PMID- 12651928 TI - Shear in flexure of fiber composites with different end supports. AB - The integrity of fiber-reinforced composite (FRC) prostheses is dependent, in part, on flexural rigidity. The object of this study was to determine if the flexure behavior of uniform FRC beams with restrained or simply supported ends and various length/depth (L/d) aspect ratios could be more accurately modeled by correcting for shear. Experimental results were compared with three analytical models. All models were accurate at high L/d ratios, but the shear-corrected model was accurate to the lowest, more clinically relevant, L/d values. In this range, more than 40% of the beam deflection was due to shear. PMID- 12651929 TI - Effects of fluoride and aluminum from ionomeric materials on S. mutans biofilm. AB - Ionomeric materials release different proportions of fluoride and aluminum. Their simultaneous effect on the acidogenicity and composition of S. mutans biofilm is unknown. Six cylindrical specimens of each material (Ketac-fil, Vitremer, Fuji Ortho LC, F-2000, and Z-100) were incubated with S. mutans GS-5 in culture media containing 5% sucrose (w/v). The media were changed daily for seven days, during which the pH and concentrations of fluoride and aluminum were determined. Furthermore, the concentrations of these ions and insoluble polysaccharide were determined in the biofilm formed at the end of the experimental period. The results showed that all the materials tested released fluoride. However, Vitremer released the highest amount of aluminum and was the most effective in reducing the acidogenicity of S. mutans biofilms. It also significantly affected both biofilm formation and composition. Thus, this study suggests that aluminum released by ionomeric materials may enhance the biological effects of fluoride. PMID- 12651930 TI - Correlations of the expression of fibroblast growth factor-2, vascular endothelial growth factor, and their receptors with angiogenesis in synovial tissues from patients with internal derangement of the temporomandibular joint. AB - Synovitis in internal derangement of the temporomandibular joint (TMJ) is accompanied by the growth of new blood vessels. Fibroblast growth factor-2 (FGF 2) and vascular endothelial growth factor (VEGF) are well-characterized angiogenic factors. The objective of this study was to elucidate the correlation between the expression of FGF-2, VEGF, and their receptors-FGF receptor-1 (FGFR 1) and VEGF receptor-1 (Flt-1)-with microvessel density in synovial tissues of the TMJ. Using an immunohistochemical technique, we examined 47 joints (45 patients) with internal derangement. Individual microvessel density was evaluated by means of the CD34 antibody, a specific endothelial marker. The correlation between the percentage of immuno-positive cells and microvessel density was evaluated. In multiple logistic regression analysis, the correlation between the percentage of Flt-1-positive cells and microvessel density was significant [p = 0.005, odds ratio = 1.071, 95% confidence interval = 1.021-1.124]. These results suggest that the expression of the VEGF/Flt-1 system is involved in angiogenesis in inflamed synovial tissue in the TMJ. PMID- 12651931 TI - Hemispheric dominance of tongue control depends on the chewing-side preference. AB - Blood-oxygenation-level-dependent (BOLD)-functional magnetic resonance imaging (fMRI) is known to be a non-invasive technique for studying human brain function. The purpose of this study was to apply BOLD-fMRI to identify brain areas responsible for producing tongue movements and their relation to chewing-side preference in 15 normal right-handed volunteers. A marked increase in BOLD signals was detected in primary sensorimotor cortices upon protrusion and in rightward and leftward tongue movements compared with at rest. In 10 subjects with an evident chewing-side preference, the BOLD signal change in the primary sensorimotor cortex was significantly greater on the side contralateral to the preferred chewing side. The results suggest that there is a relationship between hemispheric dominance and chewing-side preference in primary sensorimotor cortices responsible for tongue movements. PMID- 12651932 TI - Evidence that experimentally induced sleep bruxism is a consequence of transient arousal. AB - Spontaneous rhythmic masticatory muscle activity (RMMA) during sleep occurs more frequently following spontaneous transient micro-arousal in patients with sleep bruxism (SB) and normal controls. Here, we tested the hypothesis that an experimental arousal would be followed by an increase in RMMA. We identified RMMA on polygraphic recordings taken before and after sensory stimulation to induce experimental arousal in eight SB patients and eight matched normal subjects. The rate of experimental arousal and the level of resting electromyographic activity in masseter and suprahyoid muscles during sleep did not differ between the groups. In both, muscle tone and heart rate increased during the experimental arousal. Although post-arousal RMMA occurred in all SB patients, it was seen in only one normal subject. Moreover, tooth-grinding occurred during 71% of the evoked RMMA in SB patients. These results support the hypothesis that SB is an exaggerated form of oromotor activity associated with sleep micro-arousal. PMID- 12651933 TI - MSX1 and TGFB3 contribute to clefting in South America. AB - MSX1 and TGFB3 have been proposed as genes in which mutations may contribute to non-syndromic forms of oral clefts; however, an interaction between these genes has not been described. The present study attempts to detect transmission distortion of MSX1 and TGFB3 in 217 South American children from their respective mothers. With transmission disequilibrium test analysis, cleft lip with/without cleft palate, cleft lip with palate plus cleft palate only, and all datasets combined showed evidence of association with MSX1 (p = 0.004, p = 0.037, and p = 0.001, respectively). With likelihood ratio test analysis, "cleft lip only" showed association with MSX1 (p = 0.04) and "cleft palate only" with TGFB3 (p = 0.02). A joint analysis of MSX1 and TGFB3 suggested that there may be an interaction between these two loci to increase cleft susceptibility. These results suggest that MSX1 and TGFB3 mutations make a contribution to clefts in South American populations. PMID- 12651934 TI - Bone formation by BMP-7-transduced human gingival keratinocytes. AB - BMPs are a family of pleiotropic signaling molecules involved at various stages in the formation of bones and teeth. In addition, recombinant BMP can induce bone and dentin regeneration when applied directly to adult tissues. We have shown that fibroblasts transduced ex vivo by BMP cDNA delivered by recombinant adenoviruses induce bone formation and convert to osteoblasts upon implantation in vivo. To determine if this osteogenic capacity was limited to fibroblasts, we found that BMP-7-transduced human oral keratinocyte cells (HOKC) also formed ectopic bone. The ossicles formed by the BMP-7-transduced HOKC were smaller and more dense than those formed by BMP-7-transduced human gingival fibroblasts (HGF). Implanted HOKC were localized adjacent to the developing bone by immunocytochemical detection of keratin expression. However, no bone-like tissue formed when HOKC were implanted into diffusion chambers in vivo. We conclude that BMP-transduced HOKC secrete BMP and form bone in vivo but, unlike BMP-transduced HGF, do not transdifferentiate to osteoblasts. PMID- 12651935 TI - Regulation of osteoprotegerin gene expression in dental follicle cells. AB - Colony-stimulating factor-one (CSF-1) and parathyroid-hormone-related protein (PTHrP) down-regulate osteoprotegerin (OPG) gene expression in the dental follicle of the rat first mandibular molar. To examine this regulation at the signal transduction level, we treated cultured dental follicle cells with either phorbolmyristate acetate (PMA) or dibutyryl cyclic AMP (dbcAMP) to activate either protein kinase C (PKC) or protein kinase A (PKA). Our results demonstrate that PMA up-regulates OPG gene expression and down-regulates the expression of CSF-1 and the PTHrP receptor (PTHrP-R). Conversely, dbcAMP down-regulates OPG expression and up-regulates CSF-1 and PTHrP-R expression. Immunostaining shows that PMA also increases the steady-state levels of protein. Thus, treatment with agents that affect protein kinase activity also enhance the steady-state mRNA and protein levels of OPG, as well as decreasing the mRNA levels of CSF-1 and PTHrP R. The PKC-alpha isoform may be critical in OPG regulation because PKC-alpha gene expression is enhanced by PMA and reduced by either CSF-1 or PTHrP. PMID- 12651936 TI - Differences between tooth stimulation and capsaicin-induced neurogenic vasodilatation in human gingiva. AB - Animal experiments have shown that the application of capsaicin to oral mucosa leads to a neurogenic inflammation associated with blood flow elevations in gingivomucosal tissues. In this investigation, we measured the tooth stimulation and capsaicin-evoked blood flow responses in maxillary gingiva in humans to study whether axon-reflex-mediated vasodilatation crosses the midline of the maxilla. The vasoactive reactions were mapped by laser Doppler imaging. Unilateral stimulation of alveolar mucosa and attached gingiva by capsaicin evoked a distinct neurogenic vasodilatation in ipsilateral gingiva, which rapidly attenuated at the midline. Capsaicin stimulation of alveolar mucosa provoked clear inflammatory reactions. In contrast to capsaicin stimuli, tooth stimulation produced symmetrical vasodilatations bilaterally in the gingiva. The ipsilateral responses were significantly smaller during tooth stimulation than during capsaicin stimuli. Analysis of these data suggests that capsaicin-induced inflammatory reactions in gingivomucosal tissues do not cross the midline in the anterior maxilla. The enhanced reaction found during stimulation of alveolar mucosa indicates that alveolar mucosa is more sensitive to chemical irritants than attached gingiva. PMID- 12651937 TI - beta 2-Adrenoceptor regulation of CGRP release from capsaicin-sensitive neurons. AB - Previous studies have suggested that neurotransmitter substances from the sympatho-adrenomedullary system regulate pulpal blood flow (PBF), in part, by the inhibition of vasoactive neuropeptide release from pulpal sensory neurons. However, no study has evaluated the role of beta-adrenoceptors. We evaluated the hypothesis that activation of beta-adrenoceptors inhibits immunoreactive calcitonin gene-related peptide (iCGRP) release from capsaicin-sensitive nociceptive neurons via in vitro superfusion of bovine dental pulp. Either norepinephrine or epinephrine inhibited capsaicin-evoked iCGRP. The norepinephrine effect was blocked by the selective beta(2)-adrenoceptor antagonist, ICI 118,551, but not by pre-treatment with the selective beta(1) adrenoceptor antagonist, atenolol. In addition, application of albuterol, a selective beta(2)-adrenoceptor agonist, significantly blocked capsaicin-evoked release of iCGRP. Collectively, these studies demonstrate that activation of beta(2)-adrenoceptors in dental pulp significantly reduces exocytosis of neuropeptides from capsaicin-sensitive nociceptors. This effect may have physiologic significance in regulating PBF. Moreover, since capsaicin selectively activates nociceptors, beta(2)-adrenoceptor agonists may have clinical utility as peripherally acting therapeutics for dental pain and inflammation. PMID- 12651938 TI - Patterns of fluoro-gold entry into rat molar enamel, dentin, and pulp. AB - Permeabilities of enamel and dentin are not fully understood despite their importance for caries, restorative materials, and pulp-dentin-enamel interactions. We have found that Fluoro-Gold is useful for examining tooth permeability, and we designed studies to test the effects of aging, injury, neural function, and dentinal repair on its influx into vital rat teeth. We used fluorescence microscopy and immunocytochemistry to show that Fluoro-Gold rapidly penetrates enamel, the dentin-enamel junction, and outer dentinal acellular tubules, and then concentrates in odontoblasts, where it remains for weeks. As predicted, influx was greatest in immature teeth, and formation of reparative dentin impeded it. We expected that denervation would disrupt influx, because of neural regulation of dentinal fluid movement, but it did not. Damage to odontoblasts under injured dentin caused increased influx and efflux of Fluoro Gold. Analysis of our data suggests that permeabilities of enamel and dentin to Fluoro-Gold are age-related, inter-dependent, and regulated by odontoblasts. PMID- 12651939 TI - New animal model for studying mastication in oral motor disorders. AB - To identify the basic parameters of oral behavior in mice, we recorded the three dimensional jaw movement trajectories and masseter and digastric muscle activities in freely behaving mice eating foods of various textures. Results showed that: (1) there are characteristic jaw movement patterns for food intake and mastication; (2) the pattern in a chewing cycle may be divided into opening, closing, and protruding (power) strokes; and (3) food texture affects basic patterns of jaw movement, muscle activities, and chewing rhythms. The oral motor behavior of mice appears identical to those of other experimental animals, so mice are appropriate animal models for the study of mastication. PMID- 12651940 TI - The effect of high sugar intake on the development of periradicular lesions in rats with type 2 diabetes. AB - Diabetes mellitus is associated with depression of natural defenses against infection and increases the risk of periodontal disease. However, the effects of diabetes on periradicular tissue, which differs structurally from periodontal tissue, are not known. In this study, we evaluated the effects of type 2 diabetes on the development of periradicular lesions after exposure of the pulp in the left mandibular first molar through the occlusal surface in rats. GK rats with spontaneous non-insulin-dependent diabetes mellitus and Wistar rats (controls) received a normal laboratory diet and either water or a 30% sucrose solution. At both 2 and 4 weeks after pulp exposure, histologic analysis showed that alveolar bone resorption was most severe and the periradicular lesions were largest in diabetic rats given the sucrose solution. These results suggest that the metabolic conditions produced by type 2 diabetes enhance the development of periradicular lesions in rats. PMID- 12651941 TI - Anatomical organization of forward fiber projections from area TE to perirhinal neurons representing visual long-term memory in monkeys. AB - A number of studies have shown that the perirhinal (PRh) cortex, which is part of the medial temporal lobe memory system, plays an important role in declarative long-term memory. The PRh cortex contains neurons that represent visual long-term memory. The aim of the present study is to characterize the anatomical organization of forward projections that mediate information flow from visual area TE to memory neurons in the PRh cortex. In monkeys performing a visual pair association memory task, we conducted an extensive mapping of neuronal responses in the anteroventral part of area TE (TEav) and area 36 (A36) of the PRh cortex. Then, three retrograde tracers were separately injected into A36 and the distribution of retrograde labels in TEav was analyzed. We focused on the degree of divergent projections from TEav to memory neurons in A36, because the highly divergent nature of these forward fiber projections has been implicated in memory function. We found that the degree of divergent projection to memory neurons in A36 was smaller from the TEav neurons selective to learned pictures than from the nonselective TEav neurons. This result demonstrates that the anatomical difference (the divergence) correlates with the physiological difference (selectivity of TEav neurons to the learned pictures). Because the physiological difference is attributed to whether the projections are involved in information transmission required for memory neurons in A36, it can be speculated that the reduced divergent projection resulted from acquisition of visual long-term memory, possibly through retraction of the projecting axon collaterals. PMID- 12651942 TI - Somatic hypermutation of the B cell receptor genes B29 (Igbeta, CD79b) and mb1 (Igalpha, CD79a). AB - Somatic hypermutation (SHM), coupled to selection by antigen, generates high affinity antibodies during germinal center (GC) B cell maturation. SHM is known to affect Bcl6, four additional oncogenes in diffuse large B cell lymphoma, and the CD95Fas gene and is regarded as a major mechanism of B cell tumorigenesis. We find that mutations in the genes encoding the B cell receptor (BCR) accessory proteins B29 (Igbeta, CD79b) and mb1 (Igalpha, CD79a) occur as often as Ig genes in a broad spectrum of GC- and post-GC-derived malignant B cell lines, as well as in normal peripheral B cells. These B29 and mb1 mutations are typical SHM consisting largely of single nucleotide substitutions targeted to hotspots. The B29 and mb1 mutations appear at frequencies similar to those of other non-Ig genes but lower than Ig genes. The distribution of mb1 mutations followed the characteristic pattern found in Ig and most non-Ig genes. In contrast, B29 mutations displayed a bimodal distribution resembling the CD95Fas gene, in which promoter distal mutations conferred resistance to apoptosis. Distal B29 mutations in the cytoplasmic domain may contribute to B cell survival by limiting BCR signaling. B29 and mb1 are mutated in a much broader spectrum of GC-derived B cells than any other known somatically hypermutated non-Ig gene. This may be caused by the common cis-acting regulatory sequences that control the requisite coexpression of the B29, mb1, and Ig chains in the BCR. PMID- 12651943 TI - Regulation of hepatic fasting response by PPARgamma coactivator-1alpha (PGC-1): requirement for hepatocyte nuclear factor 4alpha in gluconeogenesis. AB - The liver plays several critical roles in the metabolic adaptation to fasting. We have shown previously that the transcriptional coactivator peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1alpha) is induced in fasted or diabetic liver and activates the entire program of gluconeogenesis. PGC-1alpha interacts with several nuclear receptors known to bind gluconeogenic promoters including the glucocorticoid receptor, hepatocyte nuclear factor 4alpha (HNF4alpha), and the peroxisome proliferator-activated receptors. However, the genetic requirement for any of these interactions has not been determined. Using hepatocytes from mice lacking HNF4alpha in the liver, we show here that PGC 1alpha completely loses its ability to activate key genes of gluconeogenesis such as phosphoenolpyruvate carboxykinase and glucose-6-phosphatase when HNF4alpha is absent. It is also shown that PGC-1alpha can induce genes of beta-oxidation and ketogenesis in hepatocytes, but these effects do not require HNF4alpha. Analysis of the glucose-6-phosphatase promoter indicates a key role for HNF4alpha-binding sites that function robustly only when HNF4alpha is coactivated by PGC-1alpha. These data illustrate the involvement of PGC-1alpha in several aspects of the hepatic fasting response and show that HNF4alpha is a critical component of PGC 1alpha-mediated gluconeogenesis. PMID- 12651944 TI - Activation-induced cytidine deaminase deaminates deoxycytidine on single-stranded DNA but requires the action of RNase. AB - The expression of activation-induced cytidine deaminase (AID) is prerequisite to a "trifecta" of key molecular events in B cells: class-switch recombination and somatic hypermutation in humans and mice and gene conversion in chickens. Although this critically important enzyme shares common sequence motifs with apolipoprotein B mRNA-editing enzyme, and exhibits deaminase activity on free deoxycytidine in solution, it has not been shown to act on either RNA or DNA. Recent mutagenesis data in Escherichia coli suggest that AID may deaminate dC on DNA, but its putative biochemical activities on either DNA or RNA remained a mystery. Here, we show that AID catalyzes deamination of dC residues on single stranded DNA in vitro but not on double-stranded DNA, RNA-DNA hybrids, or RNA. Remarkably, it has no measurable deaminase activity on single-stranded DNA unless pretreated with RNase to remove inhibitory RNA bound to AID. AID catalyzes dC --> dU deamination activity most avidly on double-stranded DNA substrates containing a small "transcription-like" single-stranded DNA bubble, suggesting a targeting mechanism for this enigmatic enzyme during somatic hypermutation. PMID- 12651946 TI - The birth and evolution of surface science: child of the union of science and technology. AB - This article is an account of the birth and evolution of surface science as an interdisciplinary research area. Surface science emanated from the confluence of concepts and tools in physics and chemistry with technological innovations that made it possible to determine the structure and properties of surfaces and interfaces and the dynamics of chemical reactions at surfaces. The combination in the 1960s and 1970s of ultra-high-vacuum (i.e., P < 10(-7) Pascal or 10(-9) Torr) technology with the recognition that electrons in the energy range from 50 to 500 eV exhibited inelastic collision mean free paths of the order of a few angstroms fostered an explosion of activity. The results were a reformulation of the theory of electron solid scattering, the nearly universal use of electron spectroscopies for surface characterization, the rise of surface science as an independent interdisciplinary research area, and the emergence of the American Vacuum Society (AVS) as a major international scientific society. The rise of microelectronics in the 1970s and 1980s resulted in huge increases in computational power. These increases enabled more complex experiments and the utilization of density functional theory for the quantitative prediction of surface structure and dynamics. Development of scanning-probe microscopies in the 1990s led to atomic resolution images of macroscopic surfaces and interfaces as well as videos of atoms moving about on surfaces during growth and diffusion. Scanning probes have since brought solid-liquid interfaces into the realm of atomic-level surface science, expanding its scope to more complex systems, including fragile biological materials and processes. PMID- 12651945 TI - Distinct roles of dopamine D2L and D2S receptor isoforms in the regulation of protein phosphorylation at presynaptic and postsynaptic sites. AB - Dopamine D2 receptors are highly expressed in the dorsal striatum where they participate in the regulation of (i) tyrosine hydroxylase (TH), in nigrostriatal nerve terminals, and (ii) the dopamine- and cAMP-regulated phosphoprotein of 32 kDa (DARPP-32), in medium spiny neurons. Two isoforms of the D2 receptor are generated by differential splicing of the same gene and are referred to as short (D2S) and long (D2L) dopamine receptors. Here we have used wild-type mice, dopamine D2 receptor knockout mice (D2 KO mice; lacking both D2S and D2L receptors) and D2L receptor-selective knockout mice (D2L KO mice) to evaluate the involvement of each isoform in the regulation of the phosphorylation of TH and DARPP-32. Incubation of striatal slices from wild-type mice with quinpirole, a dopamine D2 receptor agonist, decreased the state of phosphorylation of TH at Ser 40 and its enzymatic activity. Both effects were abolished in D2 KO mice but were still present in D2L KO mice. In wild-type mice, quinpirole inhibits the increase in DARPP-32 phosphorylation at Thr-34 induced by SKF81297, a dopamine D1 receptor agonist. This effect is absent in D2 KO as well as D2L KO mice. The inability of quinpirole to regulate DARPP-32 phosphorylation in D2L KO mice cannot be attributed to decreased coupling of D2S receptors to G proteins, because quinpirole produces a similar stimulation of [(35)S]GTPgammaS binding in wild type and D2L KO mice. These results demonstrate that D2S and D2L receptors participate in presynaptic and postsynaptic dopaminergic transmission, respectively. PMID- 12651947 TI - Structure and membrane affinity of a suite of amphiphilic siderophores produced by a marine bacterium. AB - Iron concentrations in the ocean are low enough to limit the growth of marine microorganisms, which raises questions about the molecular mechanisms these organisms use to acquire iron. Marine bacteria have been shown to produce siderophores to facilitate iron(III) uptake. We describe the structures of a suite of amphiphilic siderophores, named the amphibactins, which are produced by a nearshore isolate, gamma Proteobacterium, Vibrio sp. R-10. Each amphibactin has the same Tris-hydroxamate-containing peptidic headgroup composed of three ornithine residues and one serine residue but differs in the acyl appendage, which ranges from C-14 to C-18 and varies in the degree of saturation and hydroxylation. Although amphiphilic siderophores are relatively rare, cell associated amphiphilic siderophores are even less common. We find that the amphibactins are cell-associated siderophores. As a result of the variation in the nature of the fatty acid appendage and the cellular location of the amphibactins, the membrane partitioning of these siderophores was investigated. The physiological mixture of amphibactins had a range of membrane affinities (3.8 x 10(3) to 8.3 x 10(2) M(-1)) that are larger overall than other amphiphilic siderophores, likely accounting for their cell association. This cell association is likely an important defense against siderophore diffusion in the oceanic environment. The phylogenetic affiliation of Vibrio sp. R-10 is discussed, as well as the observed predominance of amphiphilic siderophores produced by marine bacteria in contrast to those produced by terrestrial bacteria. PMID- 12651949 TI - Structural elements of metal selectivity in metal sensor proteins. AB - Staphylococcus aureus CzrA and Mycobacterium tuberculosis NmtR are homologous zinccobalt-responsive and nickelcobalt-responsive transcriptional repressors in vivo, respectively, and members of the ArsRSmtB superfamily of prokaryotic metal sensor proteins. We show here that Zn(II) is the most potent negative allosteric regulator of czr operatorpromoter binding in vitro with the trend Zn(II)>Co(II)Ni(II), whereas the opposite holds for the binding of NmtR to the nmt operatorpromoter, Ni(II)>Co(II)>Zn(II). Characterization of the metal coordination complexes of CzrA and NmtR by UVvisible and x-ray absorption spectroscopies reveals that metals that form four-coordinate tetrahedral complexes with CzrA [Zn(II) and Co(II)] are potent regulators of DNA binding, whereas metals that form five- or six-coordinate complexes with NmtR [Ni(II) and Co(II)] are the strongest allosteric regulators in this system. Strikingly, the Zn(II) coordination complexes of CzrA and NmtR cannot be distinguished from one another by x-ray absorption spectroscopy, with the best fit a His-3-carboxylate complex in both cases. Inspection of the primary structures of CzrA and NmtR, coupled with previous functional data, suggests that three conserved His and one Asp from the C-terminal alpha5 helix donate ligands to create a four-coordinate complex in both CzrA and NmtR, with NmtR uniquely capable of expanding its coordination number in the Ni(II) and Co(II) complexes by recruiting additional His ligands from a C-terminal extension of the alpha5 helix. PMID- 12651948 TI - The retinitis pigmentosa GTPase regulator (RPGR)- interacting protein: subserving RPGR function and participating in disk morphogenesis. AB - Retinitis pigmentosa is a photoreceptor degenerative disease leading to blindness in adulthood. Leber congenital amaurosis (LCA) describes a more severe condition with visual deficit in early childhood. Defects in the retinitis pigmentosa GTPase regulator (RPGR) and an RPGR-interacting protein (RPGRIP) are known causes of retinitis pigmentosa and LCA, respectively. Both proteins localize in the photoreceptor connecting cilium (CC), a thin bridge linking the cell body and the light-sensing outer segment. We show that RPGR is absent in the CC of photoreceptors lacking RPGRIP, but not vice versa. Mice lacking RPGRIP elaborate grossly oversized outer segment disks resembling a cytochalasin D-induced defect and have a more severe disease than mice lacking RPGR. Mice lacking both proteins are phenotypically indistinguishable from mice lacking RPGRIP alone. In vitro, RPGRIP forms homodimer and elongated filaments via interactions involving its coiled-coil and C-terminal domains. We conclude that RPGRIP is a stable polymer in the CC where it tethers RPGR and that RPGR depends on RPGRIP for subcellular localization and normal function. Our data suggest that RPGRIP is also required for disk morphogenesis, putatively by regulating actin cytoskeleton dynamics. The latter hypothesis may be consistent with a distant homology between the C terminal domain of RPGRIP and an actin-fragmin kinase, predicted by fold recognition algorithms. A defect in RPGRIP encompasses loss of both functions, hence the more severe clinical manifestation as LCA. PMID- 12651950 TI - A redox switch in CopC: an intriguing copper trafficking protein that binds copper(I) and copper(II) at different sites. AB - The protein CopC from Pseudomonas syringae has been found capable of binding copper(I) and copper(II) at two different sites, occupied either one at a time or simultaneously. The protein, consisting of 102 amino acids, is known to bind copper(II) in a position that is now found consistent with a coordination arrangement including His-1, Glu-27, Asp-89, and His-91. A full solution structure analysis is reported here for Cu(I)-CopC. The copper(I) site is constituted by His-48 and three of the four Met residues (40, 43, 46, 51), which are clustered in a Met-rich region. Both copper binding sites have been characterized through extended x-ray absorption fine structure studies. They represent novel coordination environments for copper in proteins. The two sites are approximately 30 A far apart and have little affinity for the ion in the other oxidation state. Oxidation of Cu(I)-CopC or reduction of Cu(II)-CopC causes migration of copper from one site to the other. This behavior is observed both in NMR and EXAFS studies and indicates that CopC can exchange copper between two sites activated by a redox switch. CopC resides in the periplasm of Gram-negative bacteria where there is a multicopper oxidase, CopA, which may modulate the redox state of copper. CopC and CopA are coded in the same operon, responsible for copper resistance. These peculiar and novel properties of CopC are discussed with respect to their relevance for copper homeostasis. PMID- 12651951 TI - Emergency response to an anthrax attack. AB - We developed a mathematical model to compare various emergency responses in the event of an airborne anthrax attack. The system consists of an atmospheric dispersion model, an age-dependent dose-response model, a disease progression model, and a set of spatially distributed two-stage queueing systems consisting of antibiotic distribution and hospital care. Our results underscore the need for the extremely aggressive and timely use of oral antibiotics by all asymptomatics in the exposure region, distributed either preattack or by nonprofessionals postattack, and the creation of surge capacity for supportive hospital care via expanded training of nonemergency care workers at the local level and the use of federal and military resources and nationwide medical volunteers. The use of prioritization (based on disease stage andor age) at both queues, and the development and deployment of modestly rapid and sensitive biosensors, while helpful, produce only second-order improvements. PMID- 12651952 TI - Dynamic behavior of paired claudin strands within apposing plasma membranes. AB - The tight junction (TJ) strand is a linear proteinaceous polymer spanning plasma membranes, and each TJ strand associates laterally with another TJ strand in the apposing membranes of adjacent cells to form "paired" TJ strands. Claudins have been identified as the major constituents of TJ strands, and when exogenously expressed in L fibroblasts, they polymerize into paired strands, which are morphologically similar to paired TJ strands in epithelia. Here, we show that a fusion protein of GFP with claudin-1 can also form similar paired strands in L fibroblasts, allowing us to directly observe individual paired claudin strands in live cells in real time. These paired strands showed more dynamic behavior than expected; they were occasionally broken and annealed, and dynamically associated with each other in both an end-to-side and side-to-side manner. Through this behavior of individual paired claudin strands, the network of strands was reorganized dynamically. Furthermore, fluorescence recovery after photobleaching analyses revealed that claudin molecules were not mobile within paired strands. Although these observations are not necessarily representative of TJ strands per se in epithelial cells, they provide important information on the structural and kinetic properties of TJ strands in situ with significant implications for barrier function of TJs. PMID- 12651953 TI - Transcriptome dynamics of Deinococcus radiodurans recovering from ionizing radiation. AB - Deinococcus radiodurans R1 (DEIRA) is a bacterium best known for its extreme resistance to the lethal effects of ionizing radiation, but the molecular mechanisms underlying this phenotype remain poorly understood. To define the repertoire of DEIRA genes responding to acute irradiation (15 kGy), transcriptome dynamics were examined in cells representing early, middle, and late phases of recovery by using DNA microarrays covering approximately 94% of its predicted genes. At least at one time point during DEIRA recovery, 832 genes (28% of the genome) were induced and 451 genes (15%) were repressed 2-fold or more. The expression patterns of the majority of the induced genes resemble the previously characterized expression profile of recA after irradiation. DEIRA recA, which is central to genomic restoration after irradiation, is substantially up-regulated on DNA damage (early phase) and down-regulated before the onset of exponential growth (late phase). Many other genes were expressed later in recovery, displaying a growth-related pattern of induction. Genes induced in the early phase of recovery included those involved in DNA replication, repair, and recombination, cell wall metabolism, cellular transport, and many encoding uncharacterized proteins. Collectively, the microarray data suggest that DEIRA cells efficiently coordinate their recovery by a complex network, within which both DNA repair and metabolic functions play critical roles. Components of this network include a predicted distinct ATP-dependent DNA ligase and metabolic pathway switching that could prevent additional genomic damage elicited by metabolism-induced free radicals. PMID- 12651955 TI - Tuning the free-energy landscape of a WW domain by temperature, mutation, and truncation. AB - The equilibrium unfolding of the Formin binding protein 28 (FBP) WW domain, a stable three-stranded beta-sheet protein, can be described as reversible apparent two-state folding. Kinetics studied by laser temperature jump reveal a third state at temperatures below the midpoint of unfolding. The FBP free-energy surface can be tuned between three-state and two-state kinetics by changing the temperature, by truncation of the C terminus, or by selected point mutations. FBP WW domain is the smallest three-state folder studied to date and the only one that can be freely tuned between three-state and apparent two-state folding by several methods (temperature, truncation, and mutation). Its small size (28-37 residues), the availability of a quantitative reaction coordinate (phi(T)), the fast folding time scale (10s of micros), and the tunability of the folding routes by small temperature or sequence changes make this system the ideal prototype for studying more subtle features of the folding free-energy landscape by simulations or analytical theory. PMID- 12651954 TI - Genetic characterization of glucose transporter function in Leishmania mexicana. AB - Both insect and mammalian life cycle stages of Leishmania mexicana take up glucose and express all three isoforms encoded by the LmGT glucose transporter gene family. To evaluate glucose transporter function in intact parasites, a null mutant line has been created by targeted disruption of the LmGT locus that encompasses the LmGT1, LmGT2, and LmGT3 genes. This deltalmgt null mutant exhibited no detectable glucose transport activity. The growth rate of the deltalmgt knockout in the promastigote stage was reduced to a rate comparable with that of WT cells grown in the absence of glucose. deltalmgt cells also exhibited dramatically reduced infectivity to macrophages, demonstrating that expression of LmGT isoforms is essential for viability of amastigotes. Furthermore, WT L. mexicana were not able to grow as axenic culture form amastigotes if glucose was withdrawn from the medium, implying that glucose is an essential nutrient in this life cycle stage. Expression of either LmGT2 or LmGT3, but not of LmGT1, in deltalmgt null mutants significantly restored growth as promastigotes, but only LmGT3 expression substantially rescued amastigote growth in macrophages. Subcellular localization of the three isoforms was investigated in deltalmgt cells expressing individual LmGT isoforms. Using anti-LmGT antiserum and GFP-tagged LmGT fusion proteins, LmGT2 and LmGT3 were localized to the cell body, whereas LmGT1 was localized specifically to the flagellum. These results establish that each glucose transporter isoform has distinct biological functions in the parasite. PMID- 12651957 TI - Canonical and unconventional pairing schemes between bis(nucleobase) complexes of trans-a2PtII: artificial nucleobase quartets and C-H...N bonds. AB - If two nucleobases are crosslinked by trans-a(2)Pt(II), self-association via H bonding may take place either through individual bases or jointly through both bases. Due to the blockage of an acceptor site by the metal, the number of feasible pairing patterns can be reduced, and the preferred ones altered. If the metalated base pair as a whole undergoes association, base quartets can form. Various scenarios resulting from the application of guanine, hypoxanthine, and cytosine model nucleobases are discussed. Unconventional CH em leader N hydrogen bonding has been observed in several instances. PMID- 12651956 TI - Domain organization of the type 1 inositol 1,4,5-trisphosphate receptor as revealed by single-particle analysis. AB - The inositol 1,4,5-trisphosphate receptor (IP(3)R) is a tetrameric intracellular Ca(2+) channel, which mediates the release of Ca(2+) from the endoplasmic reticulum in response to many different extracellular stimuli. We present a 3D structure of the type 1 IP(3)R obtained by electron microscopy and single particle analysis that reveals its domain organization. The IP(3)R has a flower like appearance with fourfold symmetry and is made up of three distinct domains connected by slender links. By relating the organization of the structural domains to secondary-structure predictions and biochemical data we develop a model in which structural domains are mapped onto the amino acid sequence to deduce the location of the channel region and the cytoplasmic inositol 1,4,5 trisphosphate-binding and modulatory subdomains. The structure of the IP(3)R is compared with that of other tetrameric cation channels. The channel domain is similar in size and shape to its counterparts in the ryanodine receptor and the Shaker voltage-gated K(+) channel. PMID- 12651959 TI - Neuron recognition by parallel Potts segmentation. AB - Identifying neurons and their spatial coordinates in images of the cerebral cortex is a necessary step in the quantitative analysis of spatial organization in the brain. This is especially important in the study of Alzheimer's disease (AD), in which spatial neuronal organization and relationships are highly disrupted because of neuronal loss. To automate neuron recognition by using high resolution confocal microscope images from human brain tissue, we propose a recognition method based on statistical physics that consists of image preprocessing, parallel image segmentation, and cluster selection on the basis of shape, optical density, and size. We segment a preprocessed digital image into clusters by applying Monte Carlo simulations of a q-state inhomogeneous Potts model. We then select the range of Potts segmentation parameters to yield an ideal recognition of simplified objects in the test image. We apply our parallel segmentation method to control individuals and to AD patients and achieve recognition of 98% (for a control) and 93% (for an AD patient), with at most 3% false clusters. PMID- 12651958 TI - Clonal tracking of autoaggressive T cells in polymyositis by combining laser microdissection, single-cell PCR, and CDR3-spectratype analysis. AB - Clonal expansions of CD8+ T cells have been identified in muscle and blood of polymyositis patients by PCR techniques, including T cell receptor (TCR) complementarity-determining region (CDR)3 length analysis (spectratyping). To examine a possible pathogenic role of these clonally expanded T cells, we combined CDR3 spectratyping with laser microdissection and single-cell PCR of individual myocytotoxic T cells that contact, invade, and destroy a skeletal muscle fiber. First, we screened cDNA from muscle biopsy specimens by CDR3 spectratyping for expanded TCR beta chain variable region (BV) sequences. To pinpoint the corresponding T cells in tissue, we stained cryostat sections with appropriate anti-TCR BV mAbs, isolated single BV+ T cells that directly contacted or invaded a muscle fiber by laser-assisted microdissection, and amplified their TCR BV chain sequences from rearranged genomic DNA. In this way, we could relate the oligoclonal peaks identified by CDR3-spectratype screening to morphologically characterized microdissected T cells. In one patient, a large fraction of the microdissected T cells carried a common TCR-BV amino acid CDR3 motif and conservative nucleotide exchanges in the CDR3 region, suggesting an antigen driven response. In several cases, we tracked these T cell clones for several years in CD8+ (but not CD4+) blood lymphocytes and in two patients also in consecutive muscle biopsy specimens. During immunosuppressive therapy, oligoclonal CDR3-spectratype patterns tended to revert to more polyclonal Gaussian distribution-like patterns. Our findings demonstrate that CDR3 spectratyping and single-cell analysis can be combined to identify and track autoaggressive T cell clones in blood and target tissue. This approach should be applicable to other inflammatory and autoimmune disorders. PMID- 12651960 TI - Sequence information can be obtained from single DNA molecules. AB - The completion of the human genome draft has taken several years and is only the beginning of a period in which large amounts of DNA and RNA sequence information will be required from many individuals and species. Conventional sequencing technology has limitations in cost, speed, and sensitivity, with the result that the demand for sequence information far outstrips current capacity. There have been several proposals to address these issues by developing the ability to sequence single DNA molecules, but none have been experimentally demonstrated. Here we report the use of DNA polymerase to obtain sequence information from single DNA molecules by using fluorescence microscopy. We monitored repeated incorporation of fluorescently labeled nucleotides into individual DNA strands with single base resolution, allowing the determination of sequence fingerprints up to 5 bp in length. These experiments show that one can study the activity of DNA polymerase at the single molecule level with single base resolution and a high degree of parallelization, thus providing the foundation for a practical single molecule sequencing technology. PMID- 12651961 TI - Gibberella from A (venaceae) to Z (eae). AB - Gibberella species are destructive plant pathogens, although many are more familiar under their Fusarium anamorph names. The recent synthesis of phylogenetic, biological, and morphological species approaches has revitalized taxonomy of a genus that was first described almost 200 years ago. Twelve sexual species of Gibberella of agricultural importance were selected for this review to represent phylogenetic, biological, and chemical diversity of the genus. Even closely related Gibberella species can differ in reproductive mode, geographic and host distribution, plant pathogenesis, and production of toxins and other biologically active metabolites. Gibberella species have proven amenable to meiotic and molecular genetic analysis; A complete genome sequence of G. zeae should soon be available. Combining gene disruption strategies with new genomics technologies for expression profiling should help plant pathologists to understand the pathological and evolutionary significance of biological and chemical diversity in Gibberella and to identify novel strategies for disease control. PMID- 12651962 TI - Brome mosaic virus RNA replication: revealing the role of the host in RNA virus replication. AB - The replication of positive-strand RNA viruses is a complex multi-step process involving interactions between the viral genome, virus-encoded replication factors, and host factors. The plant virus brome mosaic virus (BMV) has served as a model for positive-strand RNA virus replication, recombination, and virion assembly. This review addresses recent findings on the identification and characterization of host factors in BMV RNA replication. To date, all characterized host factors facilitate steps that lead to assembly of a functional BMV RNA replication complex. Some of these host factors are required for regulation of viral gene expression. Others are needed to co-regulate BMV RNA translation and recruitment of BMV RNAs from translation to viral RNA replication complexes on the endoplasmic reticulum. Other host factors provide essential lipid modifications in the endoplasmic reticulum membrane or function as molecular chaperones to activate the replication complex. Characterizing the functions of these host factors is revealing basic aspects of virus RNA replication and helping to define the normal functions of these factors in the host. PMID- 12651963 TI - Of smuts, blasts, mildews, and blights: cAMP signaling in phytopathogenic fungi. AB - cAMP regulates morphogenesis and virulence in a wide variety of fungi including the plant pathogens. In saprophytic yeasts such as Saccharomyces cerevisiae, cAMP signaling plays an important role in nutrient sensing. In filamentous saprophytes, the cAMP pathway appears to play an integral role in vegetative growth and sporulation, with possible connections to mating. Infection-related morphogenesis includes sporulation (conidia and teliospores), formation of appressoria, infection hyphae, and sclerotia. Here, we review studies of cAMP signaling in a variety of plant fungal pathogens. The primary fungi to be considered include Ustilago maydis, Magnaporthe grisea, Cryphonectria parasitica, Colletotrichum and Fusarium species, and Erisyphe graminis. We also include related information on Trichoderma species that act as mycoparasites and biocontrol agents of phytopathogenic fungi. We point out similarities in infection mechanisms, conservation of signaling components, as well as instances of cross-talk with other signaling pathways. PMID- 12651964 TI - Quantum scattering calculations on chemical reactions. AB - This review discusses recent quantum scattering calculations on bimolecular chemical reactions in the gas phase. This theory provides detailed and accurate predictions on the dynamics and kinetics of reactions containing three atoms. In addition, the method can now be applied to reactions involving polyatomic molecules. Results obtained with both time-independent and time-dependent quantum dynamical methods are described. The review emphasises the recent development in time-dependent wave packet theories and the applications of reduced dimensionality approaches for treating polyatomic reactions. Calculations on over 40 different reactions are described. PMID- 12651965 TI - Vitamin D and its analogs as regulators of immune activation and antigen presentation. AB - It has been a little more than 20 years since the first appreciation that the biologically active hormonal form of the secosteroid vitamin D-classically categorized as a regulator of calcium/phosphorous metabolism and bone mineralization-can exert effects on cells of the immune system. Since then a substantial literature has accumulated to suggest that these effects are exerted on multiple immune cell types, are predominantly suppressive at pharmacologic levels, and are potent enough to have true therapeutic potential in the management or prevention of immune-mediated diseases. Less clear at present, however, are the physiological roles played by the vitamin D endocrine system in the regulation of normal and abnormal immune responses. In this review, an appraisal of the current understanding of vitamin D-mediated immune regulation is presented that emphasizes progress towards its clinical application as well as the manner in which emerging models of normal immune function may facilitate a more complete understanding of its physiologic significance. PMID- 12651966 TI - Dietary, evolutionary, and modernizing influences on the prevalence of type 2 diabetes. AB - An evolutionary perspective is used to elucidate the etiology of the current epidemic of type 2 diabetes estimated at 151 million people. Our primate legacy, fossil hominid, and hunting-gathering lifestyles selected for adaptive metabolically thrifty genotypes and phenotypes are rendered deleterious through modern lifestyles that increase energy input and reduce output. The processes of modernization or globalization include the availability and abundance of calorically dense/low-fiber/high-glycemic foods and the adoption of sedentary Western lifestyles, leading to obesity among both children and adults in developed and developing countries. These trends are projected to continue for a number of decades. PMID- 12651967 TI - Pain mechanisms: labeled lines versus convergence in central processing. AB - The issue of whether pain is represented by specific neural elements or by patterned activity within a convergent somatosensory subsystem has been debated for over a century. The gate control theory introduced in 1965 denied central specificity, and since then most authors have endorsed convergent wide-dynamic range neurons. Recent functional and anatomical findings provide compelling support for a new perspective that views pain in humans as a homeostatic emotion that integrates both specific labeled lines and convergent somatic activity. PMID- 12651968 TI - Fabrication, falsification and plagiarism. PMID- 12651969 TI - Physical activity: the evidence of benefit in the prevention of coronary heart disease. PMID- 12651970 TI - Assessment of endothelial damage and dysfunction: observations in relation to heart failure. PMID- 12651971 TI - Sudden, unexpected cardiac or unexplained death in England: a national survey. AB - BACKGROUND: Post-mortem examinations of adults who were apparently healthy but died suddenly and unexpectedly sometimes reveal no morphological abnormalities to explain their deaths. The frequency of such unexplained deaths in relation to other causes of sudden cardiac death is not known. AIM: To estimate the frequency of sudden unexpected cardiac or unexplained death in England. DESIGN: Prospective survey using a stratified random sample of 83 of the 132 H.M. Coroner's jurisdictions in England. METHODS: Consecutive White Caucasians, aged 16-64 years, with no medical history of cardiac disease, seen alive within 12 h of death, on whom autopsy found either a cardiac or no identifiable cause of death, were included. The coroner's officer sent a copy of the post-mortem report and a completed case registration form to the investigators, with tissue samples. RESULTS: Sixty-seven (81%) coroners participated, each maintaining prospective surveillance for 4 months. Of 692 ascertained cases, case registration forms were received for 650 (94%), post-mortem reports for 682 (99%), blood samples for 569 (82%), myocardial slices for 517 (75%) and whole hearts for 47 (7%). In cases with myocardial tissue, death was ascribed to ischaemic heart disease in 465 (82.4%). In 43.1% the ischaemia was acute, in 19.1% there was myocardial scarring but no acute ischaemia, and 20.2% had coronary atheroma only. Death was due to left ventricular hypertrophy in 32 (5.7%), to other cardiac causes in 30 (5.3%) and in 23 (4.1%) there was no clear cause. Those with cardiac causes were 81% male, median ages 55.9 (male) and 56.6 (female) years. The 23 unexplained deaths were 57% female, median ages 40.5 (male) and 54.9 (female) years. The estimated annual frequency of sudden unexpected death due to cardiac or unidentified causes, in English adults of employment age, was 11/100,000 (3481 annual deaths). DISCUSSION: In 4.1% of sudden unexpected deaths under 65 years, no cause was found. Until it becomes accepted practice to identify these cases by a name, such as Sudden Adult Death Syndrome (SADS), it will not be possible to study their aetiology systematically. PMID- 12651972 TI - Deteriorating beta-cell function in type 2 diabetes: a long-term model. AB - BACKGROUND: Type 2 diabetes is characterized by insulin resistance and the progressive loss of islet beta-cell function. Although the former is already established at diagnosis and changes little thereafter, beta-cell function continues to decline, leading to secondary failure of anti-hyperglycaemic therapies. AIM: To develop a quantitative model of the process of beta-cell function decay over time, using trial data. DESIGN: Re-analysis of published data. METHODS: The results of the Belfast Diet Study were re-analysed. Assuming patients are diagnosed at different stages in the disease process, time displacement of data was used to obtain a bi-partite spline model describing loss of insulin secretion over a 6-year period. RESULTS: The model was developed combining two phases, in which a long slow gradual loss of beta-cell function leads to a crisis in metabolic regulation, precipitating a much more rapid decay phase. This paradigm was consistent with a previous non-linear model of beta-cell mass regulation. DISCUSSION: This model may have important implications for targeting appropriate therapy to patients in each phase: delaying or avoiding full clinical type 2 diabetes in the first phase; and preventing the development of diabetic complications in the second phase. PMID- 12651973 TI - Vitamin B12 insufficiency and the risk of fetal neural tube defects. AB - BACKGROUND: Although maternal folate insufficiency is a risk factor for fetal neural tube defects (NTDs), there is controversy about whether vitamin B12 (B12) insufficiency is also associated with an increased risk of NTDs. AIM: To investigate whether low maternal B12 is associated with an increased risk of fetal NTDs. DESIGN: Systematic review. METHODS: A systematic search of Medline between 1980 and October 2002, with an examination of the citations of all retrieved studies. Studies were included that: (i) used a cohort or case-control design; (ii) included case mothers with a prior or current NTD-affected pregnancy; (iii) assessed a group of unaffected 'controls'; and (iv) measured the vitamin B12 status of all participants. RESULTS: Overall, 17 case-control studies were included, mean sample size 33 cases and 93 controls. In 5/6, mean amniotic fluid B12 concentration was significantly lower in case mothers than in controls. Of 11 that measured maternal serum or plasma B12, three observed a significantly lower mean concentration in case mothers vs. controls, while five others found a non-significant lower trend in the case group. One study observed a significantly higher mean concentration of maternal serum methylmalonic acid among the maternal cases, while another found a non-significant lower mean concentration of plasma holo-transcobalamin. Five studies estimated the risk of NTDs in relation to low B12 or B12-related metabolic markers: it was significantly increased in three studies, with a non-significant trend in the fourth. DISCUSSION: There seems to be a moderate association between low maternal B12 status and the risk of fetal NTDs. However, several design limitations, and the inclusion of few study participants, may have under-represented this. A large observational study, using reliable and valid indicators of B12 status in early pregnancy, could best assess the association between B12 insufficiency and the risk of fetal NTDs. PMID- 12651974 TI - 5,10-Methylenetetrahydrofolate reductase 677C-->T and 1298A-->C mutations are genetic determinants of elevated homocysteine. AB - BACKGROUND: Methylenetetrahydrofolate reductase (MTHFR) is one of the main regulatory enzymes of homocysteine metabolism. Elevated plasma total homocysteine (tHcy) is a major risk for cardiovascular disease. A common 677C-->T mutation in the MTHFR gene results in decreased enzymic activity, and contributes to increased plasma tHcy, in association with low plasma folate. A recently described 1298A-->C mutation in the MTHFR gene clearly reduces MTHFR activity (although to a lesser extent than the 677C-->T) but its effect on plasma tHcy levels is not yet clear. AIM: To investigate the frequency of these two MTHFR polymorphisms in a Portuguese population, and to correlate the MTHFR genotype with the biochemical phenotype at the level of homocysteine and folate concentrations. DESIGN: Prospective population survey. METHODS: We studied 117 healthy volunteers (71 females, 46 males). The 677C-->T and 1298A-->C mutations were screened by PCR-RFLP. Levels of plasma tHcy and folate, and red blood cell folate, were determined. RESULTS: The allele frequencies of the 677C-->T and 1298A-->C mutations were 0.33 and 0.28, respectively. Homozygotes for the 677C- >T mutation had significantly elevated plasma tHcy and RBC folate levels and significantly lowered plasma folate concentrations than subjects without the mutation. The 1298A-->C mutation showed a significant effect on plasma tHcy, but not on plasma folate or RBC folate levels. DISCUSSION: The observed 677T allele frequency is not consistent with the idea of a north-south gradient as previously suggested. The 1298A-->C mutation is common in Portugal. Both MTHFR mutations showed effects on plasma tHcy levels. PMID- 12651976 TI - Aetiology and pathogenesis of mood disorders. PMID- 12651975 TI - Variations in experience in obtaining local ethical approval for participation in a multi-centre study. AB - BACKGROUND: The Department of Health recently issued guidance on how Local Research Ethics Committees (LRECs) should handle an Multi-centre Research Ethics Committee (MREC)-approved application. This process is intended as a rapid standardized approval process, facilitating the execution of clinical trials. AIM: To evaluate if this guidance had led to an efficient process for obtaining local ethical approval. METHODS: Questionnaires were sent by post to Local Investigators of the 56 centres who had obtained LREC approval for the Multi centre Intrapleural Streptokinase Trial. RESULTS: Replies were received from 51 centres (91%). A total of 25 296 pieces of paper and 62 h of photocopying time were required to meet the 51 LRECs' requirements. LREC meetings ranged from weekly to bimonthly, with only 24 (47%) having a 'fast track' system in place. Applications took a median of 27 (1-90) days from submission to first being considered, with local investigators spending 3.27 (0.5-15) h on each submission. Nineteen (37%) of the local investigators felt the LREC/MREC interface did not work well and 17 (33%) were at least partly deterred from participating in future trials. DISCUSSION: The guidelines do not seem to have been implemented by all LREC committees, leading to wide variation in local experience. PMID- 12651977 TI - Research evidence and the individual patient. PMID- 12651978 TI - Metformin-associated lactic acidosis in a patient with liver disease. PMID- 12651979 TI - Itching to be treated. PMID- 12651980 TI - Danazol-induced hepatocellular carcinoma. PMID- 12651981 TI - Models have real value. PMID- 12651983 TI - Workplace social support and ambulatory cardiovascular activity in New York City traffic agents. AB - OBJECTIVE: This study examined the associations of social support from coworkers and supervisors to workday ambulatory blood pressure and heart rate. Specifically, analyses examined the association of work-related social support to the following measures: 1) baseline blood pressure and heart rate at the start of the workday, 2) blood pressure and heart rate during high stress periods, 3) blood pressure and heart rate levels throughout the workday. METHODS: Participants included male (N = 36) and female (N = 34) New York City Traffic Enforcement Agents. Mixed-model regression analyses were used to assess the effects of self-reported measures of workplace social support on workday ambulatory blood pressure. RESULTS: Workplace support was associated with workday ambulatory blood pressure levels, with the effects depending on the source of support and the gender of the participant. For women, immediate supervisor support was negatively associated with workday systolic blood pressure level, and a similar negative trend was found for workday diastolic blood pressure level. For men, coworker support was negatively associated with workday systolic blood pressure level. We found an interaction of social support with stress level (ie, baseline vs. high stress) such that immediate supervisor support was negatively associated with systolic blood pressure during high-stress conditions, but no effects were found under resting conditions. CONCLUSIONS: These findings suggest that workplace social support is associated with workday ambulatory blood pressure, especially during stressful work periods. PMID- 12651984 TI - Effects of depression on QT interval variability after myocardial infarction. AB - OBJECTIVES: Clinical depression is a risk factor for cardiac mortality in patients with coronary heart disease. High QT interval variability is a risk factor for arrhythmic events, including sudden cardiac death. The purpose of this study was to determine whether depression is associated with increased QT variability in patients recovering from myocardial infarction. METHODS: Twenty patients with major depression recovering from a recent myocardial infarction were matched with 20 nondepressed post-myocardial infarction patients on age and sex, and all underwent 24-hour Holter monitoring. RESULTS: There were no differences between groups on average heart rate, heart rate variability, or other electrocardiographic measures. However, the QT interval showed significantly greater variability in the depressed than in the nondepressed group, especially at midnight and at 6:00 AM. CONCLUSIONS: Depressed post myocardial infarction patients may be at greater risk for sudden cardiac death as a result of abnormalities in ventricular repolarization. More work is needed to determine the clinical and prognostic significance of QT variability in these patients. PMID- 12651985 TI - Cytokines in depression and heart failure. AB - OBJECTIVE: There is a convincing body of evidence linking depression, cardiovascular disease, and mortality. There is also growing evidence that depression is a risk factor for congestive heart failure (CHF) and that CHF patients with major depression have higher rates of mortality and repeat hospitalizations. Currently there are no proposed neurobiological or neuroimmune mechanisms for the comorbidity of heart failure and depression. METHODS: This review focuses on the recent literature about the role of cytokines in CHF and depression as separate conditions. This review also attempts to identify the overlapping immunological mechanisms that have a potential for future research in the pathophysiology of comorbid depression and CHF. RESULTS: Results of current studies suggest that cytokines exert deleterious effects on the heart and that soluble tumor necrosis factor (TNF) receptor 2 leads to reversal of the cardiotoxic effects of TNF, although the clinical significance of this is unclear. Major depression has been associated with alteration of various aspects of the innate immune system, including cellular components (such as microphages, neutrophils, and natural killer cells) and soluble mediators (such as acute-phase reaction proteins and cytokines). It is inconclusive whether antidepressants have immunoregulatory effects. CONCLUSIONS: The literature has not yet addressed the role of cytokines in comorbid depression and CHF. But cytokines may provide a new avenue in understanding brain-body interaction in depression and heart failure. PMID- 12651986 TI - Herpes viruses, cytokines, and altered hemostasis in vital exhaustion. AB - OBJECTIVE: Infections with herpes viruses have been implicated in the pathogenesis of atherosclerosis. We tested the hypothesis that vital exhaustion (VE) is associated with multiple herpesvirus infections, such as herpes simplex virus, varicella-zoster virus, Epstein-Barr virus, and cytomegalovirus, and with an increase in pathogen burden (ie, the aggregated seropositivity to immunoglobulin G antibodies for herpes simplex virus, varicella-zoster virus, Epstein-Barr virus, and cytomegalovirus). In addition, we examined the association of VE and pathogen burden with measures of hemostasis and inflammation. METHODS: Blood samples were drawn from 29 men with VE and 30 male control subjects, all healthy and nonsmokers, to assess serological evidence of infection and measures of hemostasis and inflammation. RESULTS: VE is associated with a relatively high pathogen burden, altered hemostasis, and higher levels of cytokines, such as interleukin-6. Across all subjects, a relatively high pathogen burden was also associated with altered hemostasis but not with increased cytokine levels. The interaction of VE with pathogen burden revealed significant linear increases in measures of hemostasis and inflammation. Finally, immunoglobulin G antibody titer levels of individual herpesvirus infections were not associated with hemostatic measures or with cytokines. CONCLUSIONS: We conclude that stress-related alterations in hemostasis and inflammation are not necessarily linked to one particular herpesvirus infection but rather to an increase in aggregated seropositivity to herpesvirus infections. PMID- 12651987 TI - Do depressive symptoms increase the risk for the onset of coronary disease? A systematic quantitative review. AB - OBJECTIVE: The objectives of this study were to systematically review the recent studies of the contribution of depression to the onset of coronary disease and to estimate the magnitude of the risk posed by depression for onset of coronary disease. METHOD: We searched MEDLINE (1966-2000), PsychInfo (1967-2000), and cross references and conducted informal searches for all community studies of depression symptoms in samples with no clinically apparent heart disease at baseline. From these studies we selected all published cohort studies of 4 years or more follow-up that controlled for other major coronary disease risk factors and reported relative risks (or a comparable measure) of baseline depression for the onset of coronary disease. Following methods for the meta-analysis of epidemiologic studies, we used a random-effects model to estimate the combined overall relative risk. RESULTS: Ten studies met our inclusion criteria. Relative risks ranged from 0.98 to 3.5. Nine studies reported significantly increased risk, including two with mixed results; one study reported no increased risk. The combined overall relative risk of depression for the onset of coronary disease was 1.64 (95% CI = 1.41-1.90). CONCLUSIONS: This quantitative review suggests that depressive symptoms contribute a significant independent risk for the onset of coronary disease, a risk (1.64) that is greater than the risk conferred by passive smoking (1.25) but less than the risk conferred by active smoking (2.5). Future prospective community studies should examine the effect of severity and duration of depressive symptoms and disorders on the risk for the onset of coronary disease. PMID- 12651988 TI - Chronic insomnia and immune functioning. AB - OBJECTIVE: The goal of this study was to investigate whether clinical insomnia is associated with immune alterations by comparing immune functioning between patients with chronic insomnia and good sleepers. METHODS: The good sleepers group was composed of 19 adults with a regular sleep schedule and no complaint of sleep disturbances. The insomnia group was composed of 17 adults meeting criteria for a chronic primary insomnia disorder. Peripheral blood samples were taken at the interview (time 1) and before the second night of polysomnographic assessment (time 2) for immune measures, including enumeration of blood cell counts (ie, white blood cells, monocytes, lymphocytes, and CD3+, CD4+, CD8+, and CD16+/CD56+ cells), natural killer cell activity, and cytokine production (ie, interleukin 1beta, interleukin-2, and interferon gamma). RESULTS: Significant between-group differences were observed for CD3+, CD4+, and CD8+ cells, with higher levels found in good sleepers. In addition, a significant group-by-time interaction was found on monocyte counts. Although this was the only significant interaction effect observed, between-group differences were greater at time 2. CONCLUSIONS: This study suggests that chronic insomnia is associated with some immune alterations. More research is needed to determine the clinical significance of these findings. PMID- 12651989 TI - Exaggerated electrodermal startle responses after intracardiac shock discharges in patients with implanted cardioverter defibrillators. AB - OBJECTIVE: We studied whether repetitive intracardiac shock discharges of implantable cardioverter defibrillators (ICDs) provoke an enduring enhancement of startle responses. METHODS: The study population comprised 134 patients with an ICD. Among those, 67 patients had experienced shock delivery. Thirty-five patients had received five or more shocks. We used the startle reflex paradigm, which consisted of 15 acoustic stimuli (95 dB, 1000 Hz, 500 ms duration). Skin conductance response was measured using a constant 0.5 V through 8-mm electrodes placed on each subject's nondominant palm. Response magnitude was calculated by subtracting the baseline response level of the 2 seconds immediately preceding tone onset from the maximum response level within 1 to 4 seconds after tone onset. The left orbicularis oculi electromyogram (EMG) response was calculated by subtracting the mean EMG level during the 2 seconds immediately preceding tone onset from the highest EMG level measured within 40 to 200 ms after tone onset. Habituation was defined by the response slope of the regression equation and by the number of trials required to reach the nonresponse criterion. RESULTS: Although EMG response measures of magnitude and habituation failed to yield differences between study groups, patients who had experienced five or more ICD shocks exhibited a significantly larger skin conductance response magnitude in comparison to the patients who had experienced fewer than five shocks (median, interquartile range: 0.364, 0.209-0.618 vs. 0.512, 0.375-0.791; Mann-Whitney U test, p =.007). Poorer habituation in the group with five or more shocks in comparison with the low shock group was confirmed both by the number of trials needed to reach the nonresponse criterion (median, interquartile range: 10, 5-14 vs. 5, 2-13; p =.003) and by the response slope (median, interquartile range: 0.209, 0.116-0.274 vs. 0.262, 0.181-0.332; p =.008). After controlling for potential confounding factors (age, anxiety, aversiveness of stimuli, time since last shock experience, and use of beta-adrenoceptor antagonists), intracardiac shock discharges had the strongest impact on augmented skin conductance response magnitude (adjusted odds ratio = 3.0, 95% confidence interval = 1.3-7.2, p =.01) and impaired habituation (adjusted odds ratio = 2.8, 95% confidence interval = 1.2-6.3; p =.015). CONCLUSIONS: Intracardiac shock discharges are associated with augmented skin conductance responses and slower habituation, indicating sustained sympathetic arousability, which is presumably centrally mediated. PMID- 12651990 TI - Depression predicts revascularization procedures for 5 years after coronary angiography. AB - OBJECTIVE: Depression has been reported to increase cardiac event rates and functional impairment in patients with coronary disease. This article describes the impact of depression on subsequent healthcare utilization for such patients. METHODS: One hundred ninety-eight health maintenance organization patients with stable coronary disease were interviewed after elective angiography using a structured psychiatric diagnostic scale. Cardiac events, hospitalizations, procedures, and costs were monitored for the next 5 years through automated data. Subjects were classified at the time of angiography by modified DSM-IV criteria into those with major, minor, and no depression. RESULTS: In univariate analyses, the no depression group (N = 136) was most likely to receive coronary artery bypass grafting (CABG) (61% vs. 36% in the major depression group vs. 27% in the minor depression group, p =.001), and the major depression group (N = 25) was most likely to receive percutaneous transluminal coronary angioplasty (PTCA) (44% vs. 14% in the minor depression group vs. 24% in the no depression group). The minor depression group (N = 37) was least likely to be hospitalized for cardiac reasons during follow-up (54% vs. 80% in the major depression group vs. 80% in the no depression group, p =.005). Five-year rates of myocardial infarction and death did not differ significantly between groups. Proportional hazard models showed that those in the depression groups differed in time from catheterization to CABG (chi2(2) = 11.9, p =.003) and time to PCTA (chi2(2) = 7.74, p =.02) after controlling for relevant covariates. Median regression showed that patients with no depression had higher costs during the first year but tended to have lower costs in years 2 through 5 than patients with minor or major depression. CONCLUSIONS: Depression status at angiography is associated with the need for revascularization and total healthcare costs for the following year. PMID- 12651991 TI - Association between the menses and suicide attempts: a replication study. AB - OBJECTIVE: In our prior (first) study, suicide attempts were associated with the menses. The main hypothesis of this replication (second) study is that the proportion of suicide attempters during the menses is significantly higher than the expected probability in the general population. METHODS: Using the same methodology and setting, this replication naturalistic study included 120 fertile female suicide attempters with regular menstrual cycles during a 1-year period in the emergency room of a general hospital in Madrid, Spain, serving a catchment area of 500,000 people. RESULTS: The significant increase in probability of attempting suicide during the menses for the first study was 1.61; for the second study, 1.72; and for both studies combined, 1.68 (95% confidence interval, 1.27 2.09). Thus, using the combined results, the probability of attempting suicide during the menses was 1.68 times higher than the overall probability of attempting suicide for any fertile women. In the catchment area, the population rate of fertile women arriving at the hospital after a suicide attempt was 166 per 100,000. The probability for women during the menses arriving at the hospital after a suicide attempt was significantly higher, 279 per 100,000 (1.68 x 166 per 100,000). The 95% confidence interval was 211 to 347 per 100,000. CONCLUSIONS: Despite inherent limitations, this naturalistic study replicates a small but significant increase of suicide attempts during the menses. PMID- 12651992 TI - Innate secretory immunity in response to laboratory stressors that evoke distinct patterns of cardiac autonomic activity. AB - OBJECTIVE: Most infections begin at mucosal surfaces. These surfaces are covered by the secretory proteins of the exocrine glands (eg, the salivary, respiratory, and gastrointestinal glands), which provide a first line of innate defense. The release of these secretory proteins is under neuroendocrine control and thus, in theory, sensitive to modulation by psychosocial stress. This was empirically tested by measuring the salivary secretion of cystatin S, lactoferrin, alpha amylase, the mucins MUC5B and MUC7, and total salivary protein in response to stressors known to evoke distinct patterns of cardiac autonomic activity. METHODS: Thirty-two undergraduate volunteers were each subjected to two laboratory stressors and a control condition. Stressors were an active coping memory test and a passive coping video presentation showing surgical procedures. In the control condition participants viewed a didactic video presentation. RESULTS: The stressors evoked the expected distinct patterns of cardiac autonomic activity. The memory test produced a strong increase in sympathetic activity (evidenced by a shortened preejection period), and a decrease in cardiac parasympathetic activity (evidenced by a decrease in heart rate variability). This active coping response was associated with an enhanced secretion ( microg/min, controlling for salivary flow rate) of MUC7, lactoferrin, alpha amylase, and total salivary protein. Conversely, the surgical video produced an increase in cardiac vagal tone and a modest increase in sympathetic activity. This passive coping response was associated with an enhanced secretion of all proteins studied. These secretory responses were generally larger than the secretory responses during the active coping memory test. Correlation analyses indicated that for both stressors autonomic and cardiovascular reactivity was positively associated with an enhanced and prolonged secretory activity. CONCLUSIONS: Stress-induced modulation of innate secretory immunity may be a contributing factor in the observed relationship between stress and susceptibility to infectious diseases. We further propose a more differentiated approach to acute stress by distinguishing among stressors with distinct autonomic nervous system effects. PMID- 12651993 TI - Role of stress, arousal, and coping skills in primary insomnia. AB - OBJECTIVE: Although stress is often presumed to cause sleep disturbances, little research has documented the role of stressful life events in primary insomnia. The present study examined the relationship of stress and coping skills, and the potential mediating role of presleep arousal, to sleep patterns in good sleepers and insomnia sufferers. METHODS: The sample was composed of 67 participants (38 women, 29 men; mean age, 39.6 years), 40 individuals with insomnia and 27 good sleepers. Subjects completed prospective, daily measures of stressful events, presleep arousal, and sleep for 21 consecutive days. In addition, they completed several retrospective and global measures of depression, anxiety, stressful life events, and coping skills. RESULTS: The results showed that poor and good sleepers reported equivalent numbers of minor stressful life events. However, insomniacs rated both the impact of daily minor stressors and the intensity of major negative life events higher than did good sleepers. In addition, insomniacs perceived their lives as more stressful, relied more on emotion-oriented coping strategies, and reported greater presleep arousal than good sleepers. Prospective daily data showed significant relationships between daytime stress and nighttime sleep, but presleep arousal and coping skills played an important mediating role. CONCLUSIONS: The findings suggest that the appraisal of stressors and the perceived lack of control over stressful events, rather than the number of stressful events per se, enhance the vulnerability to insomnia. Arousal and coping skills play an important mediating role between stress and sleep. The main implication of these results is that insomnia treatments should incorporate clinical methods designed to teach effective stress appraisal and coping skills. PMID- 12651994 TI - Comorbid illness in women with chronic fatigue syndrome: a test of the single syndrome hypothesis. AB - OBJECTIVE: Evidence of comorbidity among unexplained illness syndromes raises the possibility that all are variants of a single functional disorder, leading some to suggest that separate case definitions for chronic fatigue syndrome (CFS), fibromyalgia (FM), and multiple chemical sensitivity (MCS) may be unnecessary. Our objective was to determine whether discrete diagnostic labels provide useful information about physical functioning, symptom severity, and risk of psychiatric illness. METHODS: The sample consisted of 163 consecutive female referrals with CFS enrolled at a tertiary clinic. Each participant was retrospectively assigned to one of four groups: CFS only, CFS/FM, CFS/MCS, and CFS/FM/MCS. At enrollment, participants gave their history, underwent a physical examination and a standardized psychiatric interview (Diagnostic Interview Schedule), and answered self-report questionnaires. RESULTS: Additional unexplained syndromes were prevalent: 37% met criteria for FM, and 33% met criteria for MCS. With the exception of FM-related pain and disability, there were few differences between the CFS only and CFS with comorbid illness groups. Patients with additional illness were more likely to have major depression and a higher risk of psychiatric morbidity compared with patients in the CFS only group (p <.01). Rates of lifetime depression increased from 27.4% in the CFS only group to 52.3% in the CFS/FM group, 45.2% in the CFS/MCS group, and 69.2% in the CFS/FM/MCS group. CONCLUSIONS: The prevalence of comorbid illness in the present CFS sample and the failure to find widespread differences in symptom severity can be seen as support for the single syndrome hypothesis. On the other hand, the existence of discrete syndromes could not be ruled out because of reliable differences between CFS and CFS/FM. Increasing comorbidity was associated with a corresponding increase in risk of major depression. PMID- 12651995 TI - Social support and experimental pain. AB - OBJECTIVE: The purpose of this experimental study was to supplement and expand on clinical research demonstrating that the provision of social support is associated with lower levels of acute pain. METHODS: Undergraduates (52 men and 49 women) performed the cold pressor task either alone or accompanied by a friend or stranger who provided active support, passive support, or interaction. Pain perception was measured on a 10-point scale. RESULTS: Participants in the active support and passive support conditions reported less pain than participants in the alone and interaction conditions, regardless of whether they were paired with a friend or stranger. CONCLUSIONS: These data suggest that the presence of an individual who provides passive or active support reduces experimental pain. PMID- 12651996 TI - Sex differences and incentive effects on perceptual and cardiovascular responses to cold pressor pain. AB - OBJECTIVE: Sex differences in pain perception have been widely reported, with women typically displaying greater pain sensitivity than men, but the mechanisms underlying these differences remain unclear. One possible explanation suggests that men are more motivated to tolerate and suppress expressions of pain because of the masculine sex role, whereas the feminine sex role encourages pain expression and produces lower motivation to tolerate pain among women. METHODS: To examine the influence of motivation on perceptual and cardiovascular responses to pain among women and men, different levels of monetary incentive (high vs. low incentive) were provided to a group of 81 healthy young adults undergoing the cold pressor pain procedure. It was anticipated that men would have greater endogenous motivation and would therefore be less affected by the external incentive. RESULTS: Men had higher pain thresholds and tolerances and lower pain ratings than women, but the incentive condition produced no significant effect on pain responses. Resting blood pressure was positively correlated with pain tolerance among the low incentive group, whereas blood pressure reactivity to the cold pressor predicted pain tolerance in the high incentive group. CONCLUSIONS: Thus, monetary incentive did not influence pain responses, but the relationship between cardiovascular measures and pain responses was influenced by the incentive manipulation. Potential explanations for the observed results are presented, and the implications for applying the biopsychosocial model to pain research are discussed. PMID- 12651997 TI - Test of an interpersonal model of hypochondriasis. AB - OBJECTIVE: The aim of this study was to test the interpersonal model of hypochondriasis proposed by Stuart and Noyes. According to this model, hypochondriasis is associated with insecure attachment that in adults gives rise to abnormal care-seeking behavior. Such behavior is associated with interpersonal difficulties and strained patient-physician relationships. METHODS: One hundred sixty-two patients attending a general medicine clinic were interviewed and asked to complete self-report measures. Instruments included the Whiteley Index of Hypochondriasis, Somatic Symptom Inventory, Relationship Scales Questionnaire, Inventory of Interpersonal Problems, NEO Five-Factor Index, and measures of physician-patient interaction. The Structured Diagnostic Interview for DSM-III-R Hypochondriasis was also administered. RESULTS: Hypochondriacal and somatic symptoms were positively correlated with all of the insecure attachment styles, especially the fearful style. These same symptoms were positively correlated with self-reported interpersonal problems and negatively correlated with patient ratings of satisfaction with, and reassurance from, medical care. Hypochondriacal and somatic symptoms were also positively correlated with neuroticism. CONCLUSIONS: The findings indicate that hypochondriacal patients are insecurely attached and have interpersonal problems that extend to and include the patient physician relationship. These data support the proposed interpersonal model of hypochondriasis. PMID- 12651998 TI - Alexithymia and dream recall upon spontaneous morning awakening. AB - OBJECTIVE: The issue of a quantitative and qualitative impoverishment of dream activity in alexithymic subjects was assessed by analysis of the 14-day dream reports of two groups of accurately selected alexithymic and nonalexithymic subjects. METHODS: Ten alexithymic and 10 nonalexithymic women were selected from a larger sample of 160 undergraduate students. The transcriptions of their audio recorded dream reports on 14 consecutive morning awakenings were compared with regard to their length and emotional content. Self-ratings obtained from sleep and dream diaries were further considered to assess between-groups differences in dream recall frequency and in the emotional valence, vividness, and bizarreness of dreams. RESULTS: Dream recall frequency and mean length of dream reports were lower in alexithymic than in nonalexithymic subjects. There were no significant between-groups differences in emotional valence, vividness, bizarreness, and emotions scored according to the Hall and Van de Castle coding system. CONCLUSIONS: The results suggest a general difficulty of alexithymics in accessing (recalling) their dreams. PMID- 12651999 TI - Social situation of expectant mothers and alexithymia 31 years later in their offspring: a prospective study. AB - OBJECTIVE: We studied the association between alexithymia in adulthood and social situation of the child's family at the time of the child's birth. METHODS: The study forms part of the prospective Northern Finland 1966 Birth Cohort Project. The original material consisted of all 12,058 live-born children in the provinces of Lapland and Oulu in Finland with an expected delivery date during 1966. The material represents 96% of all births in the region. In 1997 a 31-year follow-up study was made on part of the initial sample. The 20-item version of the Toronto Alexithymia Scale (TAS-20) was given to 5983 participants of the field study. Of them, 85% returned the questionnaire properly completed. Factors indicative of the social situation of the child's family were already gathered during pregnancy and the time of birth from the mothers, thus eliminating recall bias. RESULTS: Alexithymia was more common in subjects from rural than urban dwelling areas. Unwanted children or subjects born into families with many children also exhibited an association with alexithymia in adulthood. CONCLUSIONS: Maternal social situation and environment predict alexithymia in the offspring. PMID- 12652000 TI - Compliance with saliva sampling protocols: electronic monitoring reveals invalid cortisol daytime profiles in noncompliant subjects. AB - OBJECTIVE: Ambulatory saliva collections for subsequent analysis of free cortisol levels are now frequently applied to measure adrenocortical activity in healthy subjects and patient populations. Despite the prime importance of accurate timing of saliva collection outside the laboratory, no data are available on the compliance of study participants following a given sampling protocol. This study investigated how accurately subjects adhered to the instructions to collect six saliva samples throughout 1 day. METHODS: Subjects were instructed to collect six saliva samples throughout 1 day (directly after awakening, 30 minutes after awakening, 11 AM, 3 PM, 8 PM, 10 PM). Objective compliance was measured using an electronic monitoring device given to the subjects either with ("informed" N = 23) or without ("noninformed" N = 24) their knowledge of the nature of the device. Data on subjective compliance were obtained by self-report. RESULTS: Thirty-one subjects (74%) were found to comply with the sampling instructions, and 11 (26%) failed at least once to obtain the saliva sample at the correct time of day. Nine of the 11 noncompliant subjects (82%) had two or more noncompliant samples. Fifty-five percent (6 of 11) of the noncompliant subjects took sample 2 outside the sampling window. The circadian cortisol profile differed significantly between compliant and noncompliant subjects (F = 7.98, p =.007). The most important effect of compliance was seen in the rise of cortisol at awakening. Compliant subjects showed a robust increase, whereas noncompliant individuals had only minimal changes from baseline at 30 minutes after awakening (t = 2.89, p =.007). Thus the steepness of the circadian cortisol decline was greater for compliant subjects (t = 2.10, p =.043). Furthermore, the informed group adhered more closely to the sampling protocol than the noninformed subjects (p =.001). Self-reported compliance also differed significantly between study groups (p =.03). In the noninformed group, self-reported sampling accuracy was significantly higher than objectively measured compliance (p =.03); the two measures were similar in the informed group (p = NS). CONCLUSIONS: A significant number of subjects did not obtain saliva samples reliably in an ambulatory setting. This can partially invalidate the cortisol results and mask potential differences between subject groups of interest. We therefore recommend the use of electronic monitoring devices or other suitable methods and that study participants be informed about the device when ambulatory saliva collection is performed. PMID- 12652001 TI - Components of the diurnal cortisol cycle in relation to upper respiratory symptoms and perceived stress. AB - OBJECTIVE: The purpose of this study was to investigate the relationship between the diurnal pattern of salivary free cortisol to perceived stress and susceptibility to symptoms of upper respiratory illness (URI). METHODS: Salivary free cortisol concentration was determined in 34 healthy participants (students) at eight time points, synchronized to awakening, on 2 consecutive days. Participants completed a standard questionnaire to assess perceived stress and subsequently kept a daily record of social proximity and symptoms of upper respiratory illness for 2 weeks. RESULTS: Participants characterized by consistently larger awakening responses went on to report significantly more URI symptoms. Participants with less pronounced diurnal decline (flatter profiles) reported fewer URI symptoms. The two cortisol components were themselves related and interacted such that participants high on an interactive vector reported approximately three times more URI symptoms than other participants. The URI associated cortisol components (dynamic changes) were not related to perceived stress, but underlying cortisol secretory activity (overall levels) in the first 45 minutes after awakening was. Dynamic components were, however, related to a social proximity measure, which in turn was related to URI symptoms. Proximity and the interactive cortisol vector together explained a substantial (28%) percentage of the variance in URI symptom reports. The cortisol vector independently and significantly explained 12% of the variation; the proximity measure independently and nonsignificantly contributed 6% of the variation. CONCLUSIONS: URI symptoms were associated with two related dynamic components of the cortisol cycle as determined by synchronization to awakening, whereas stress was related to a measure of overall secretory activity. PMID- 12652002 TI - Clinical issues in animal models of stroke and rehabilitation. PMID- 12652003 TI - Animal models of focal and global cerebral ischemia. AB - The use of appropriate animal models is essential to predict the value and effect of therapeutic approaches in human subjects. Focal (stroke) and global (cardiac arrest) cerebral ischemia represents diseases that are common in the human population. Stroke and cardiac arrest, which are major causes of death and disability, affect millions of individuals around the world and are responsible for the leading health care costs of all diseases. Understanding the mechanisms of injury and neuroprotection in these diseases is critical if we are ever to learn new target sites to treat ischemia. There are many animal models available to investigate injury mechanisms and neuroprotective strategies. This review summarizes many (but not all) small and large animal models of focal and global cerebral ischemia and discusses their advantages and disadvantages. PMID- 12652004 TI - Models of focal cerebral ischemia in the nonhuman primate. AB - Ischemic stroke is a uniquely human disease syndrome. Models of focal cerebral ischemia developed in nonhuman primates provide clinically relevant platforms for investigating pathophysiological alterations associated with ischemic brain injury, microvascular responses, treatment responses, and clinically relevant outcomes that may be appropriate for ischemic stroke patients. A considerable number of advantages attend the use of nonhuman primate models in cerebral vascular research. Appropriate development of such models requires neurosurgical expertise to produce single or multiple vascular occlusions. A number of experimentally and clinically accessible outcomes can be measured, including neurological deficits, neuron injury, evidence of non-neuronal cell injury, infarction volume, real-time imaging of injury development, vascular responses, regional cerebral blood flow, microvascular events, the relation between neuron and vascular events, and behavioral outcomes. Nonhuman primate models of focal cerebral ischemia provide excellent opportunities for understanding the vascular and cellular pathophysiology of cerebral ischemic injury, which resembles human ischemic stroke, and the appropriate study of pharmacological interventions in a human relevant setting. PMID- 12652005 TI - Perspectives on reperfusion-induced damage in rodent models of experimental focal ischemia and role of gamma-protein kinase C. AB - Ischemic stroke represents the leading cause of death and disability among elderly people. Most stroke survivors are left with lifelong disability. With the exception of tissue-type plasminogen activator (t-PA), no effective therapy exists for the management of acute stroke. Understanding the role of various extrinsic and intrinsic pathogenic factors of ischemic damage represents a prime objective of ongoing stroke research. An important variable affecting stroke outcome is the presence or absence of reperfusion (recanalization of the occluded vessel) following an ischemic event. It appears that early reperfusion after a stroke is beneficial and capable of reversing the majority of ischemic dysfunctions. However, in some instances, late reperfusion may contrarily trigger deleterious processes and lead to more ischemic damage. Examples of ischemia/reperfusion damage using an experimental model of focal ischemia in rodents are provided, along with evidence that the brain-enriched gamma-isoform of protein kinase C may represent an important mediator of reperfusion-induced brain injury in mutant mice. PMID- 12652006 TI - Neuroprotective effects on somatotopic maps resulting from piracetam treatment and environmental enrichment after focal cortical injury. AB - Acute and chronic postlesion reorganization of the cortical maps was examined in adult rats using electrophysiological mapping of the forepaw area in the primary somatosensory cortex. Recordings were made before and after (first 12 hr and 3 wk) induction of a focal thermal-ischemic lesion to a restricted part of the forepaw area. The influence of an anti-ischemic substance (piracetam) and housing in an enriched environment (EE) or impoverished environment (IE) on the organization of the spared regions of the cortical maps adjacent to the lesion was investigated. The results revealed (1) a gradual expansion of the injured zone and a cellular loss that were smaller in the piracetam-treated (PT) rats than in the placebo (PL) rats; (2) a better preservation of the somatotopic organization and the neuronal responsiveness in the maps of the PT rats during the first 12 hr after the lesion; (3) a gradual compression and fragmentation of the remaining forepaw map over the first 3 postlesion wk. These changes were found in all PL rats, with the most detrimental effects in the animals exposed to an IE. In the PT-EE animals, a contrasting substantial preservation of the map was observed. (4) Cortical responsiveness was diminished in the PL rats, whatever the environment, and in the PT-IE rats; but it was not significantly affected in the PT-EE animals. The findings demonstrate the protective function of acute piracetam treatment on electrophysiological properties of cortical neurons within the peri-infarct tissue and highlight the neuroprotective effects of an environmental therapy combined with the piracetam treatment during the weeks after ischemic damage. PMID- 12652007 TI - Model of recovery of locomotor ability after sensorimotor cortex injury in rats. AB - Animal models of locomotor recovery after brain injury provide tools for understanding the basic neurobiological processes that may underlie recovery after stroke in humans. Measurement of the ability of rats to traverse a narrow elevated beam has proven to be a particularly useful test of locomotor function. Repeated measurement of this behavior over time provides a simple method for quantifying the rate and degree of a rat's locomotor recovery after sensorimotor cortex injury and constitutes a tool for studying its mechanisms and possible treatment strategies. The model has proven particularly useful in predicting the effects of drugs on poststroke recovery in humans. PMID- 12652008 TI - Experimental focal ischemic injury: behavior-brain interactions and issues of animal handling and housing. AB - In experimental neurological models of brain injury, behavioral manipulations before and after the insult can have a major impact on molecular, anatomical, and functional outcome. Investigators using animals for preclinical research should keep in mind that people with brain injury have lived in, and will continue to live in, an environment that is far more complex than that of the typical laboratory rodent. To yield more reliable and relevant behavioral assessment, it may be appropriate in some cases to house animals in environments that allow for motor enrichment and to handle animals in ways that promote tameness. Experience can affect mechanisms of plasticity and degeneration beneficially or adversely. Behavioral interventions that have been found to modulate postinjury brain events are reviewed. The timing and interaction of biological and motor therapies and the potential contribution of experience-dependent and drug-induced trophic factor expression are discussed. PMID- 12652009 TI - Importance of behavioral manipulations and measures in rat models of brain damage and brain repair. AB - The relevance of careful behavioral measures and manipulations in animal research on neural plasticity and brain damage has become increasingly clear. Recent research in adult rats indicates that an understanding of neural restructuring after brain damage requires an understanding of how it is influenced by postinjury behavioral experiences. Other research indicates that optimizing pharmacological and other treatments for brain damage may require their combination with rehabilitative training. Assessing the efficacy of a treatment approach in animal models requires the use of sensitive behavioral measures of functional outcome. In research on restorative plasticity after brain damage, procedures for handling and housing rats should promote the quality of behavioral measures and manipulations. PMID- 12652010 TI - Assessment of cognitive and motor deficits in a marmoset model of stroke. AB - The Stroke Therapy Academic Industry Roundtable noted the need for standardized, well-accepted primate models of stroke to help develop both neuroprotective and restorative therapies. One primate model has been developed using the marmoset, a small New World species of monkey, in which long-term functional deficits can be assessed. The surgery and postoperative care of the animals is described, as well as the behavioral tests used to quantify the postoperative disability. The types of deficits seen are illustrated by reference to some of the findings with neuroprotective treatments. Nevertheless, the long-term nature and consistency of the motor deficits make this model ideal for assessing the worth of restorative therapies. PMID- 12652011 TI - A squirrel monkey model of poststroke motor recovery. AB - Nonhuman primate models of poststroke recovery have become increasingly rare primarily due to high purchase and maintenance costs and limited availability of nonhuman primate species. Despite this obstacle, nonhuman primate models may offer important advantages over rodent models for understanding many of the brain's mechanisms for self-repair due to greater similarity in cortical organization to humans. Since the mid-1990s, surgical, neurophysiological, and neuroanatomical methods have been developed to understand structural and functional remodeling of the cerebral cortex after an ischemic event, such as occurs in stroke. These methods require long surgical procedures and entail constant physiological monitoring. With careful attention to intraoperative and postsurgical monitoring, these procedures can be repeated multiple times in individual monkeys without untoward events. This model provides a statistically powerful approach for tracking brain plasticity in the ensuing weeks and months after a stroke-like injury, reducing the number of animals required for individual experiments. This methodology is described in detail, and many of the resulting findings that are relevant for understanding stroke recovery and the effects of rehabilitative and pharmacotherapeutic interventions are summarized. PMID- 12652012 TI - Rheological behaviour of red blood cells in beta and deltabeta thalassemia trait. AB - In major and intermediate thalassemia a decrease in erythrocyte deformability and increased erythrocyte aggregability has been described, but few studies have dealt with the question of rheological red blood cell behaviour in minor beta and deltabeta thalassemia carriers, mostly in deltabeta, because it is a less common entity. To ascertain whether there are differences in red blood cell behaviour between minor thalassemia and controls and between both types of thalassemia trait beta and deltabeta, we determined erythrocyte deformability and aggregability in 30 beta and 30 deltabeta trait carriers diagnosed both with conventional methods and globin gene analysis, and in 40 age- and sex-matched controls. Erythrocyte deformability determined by means of the Rheodyn SSD showed a statistically significant lower Elongation Index (EI) at all the shear stresses tested in both thalassemic groups compared with controls (p<0.001). Minor beta thalassemia carriers showed lower EI than deltabeta carriers (p<0.001). Erythrocyte aggregability measured with the Myrenne aggregometer was significantly lower in both thalassemic groups than in controls (p<0.001), although no significant differences could be observed between both thalassemic groups. The rheological alterations found in thalassemia carriers are in part due to microcytosis, hypochromia and the morphological changes that characterize this kind of anaemia. The less altered deformability found in deltabeta carriers, is in agreement with the fact that it deals with a more benign trait. PMID- 12652013 TI - Influence of a parenteral fish-oil preparation (Omegaven) on erythrocyte morphology and blood viscosity in vitro. AB - In most studies fish-oil had been administered by enteral route. Recently an parenteral marine fish-oil emulsion has been developed. With an intravenous infusion an immediate effect on the membrane lipid composition of circulating blood cells and endothelial cells may be expected. In this study we assessed the acute effects of a parenteral fish-oil emulsion on blood rheology and erythrocyte morphology in vitro. Blood from 15 healthy young volunteers was incubated in vitro with increasing concentrations of a parenteral fish-oil emulsion (Omegaven) at 37 degrees C for different time periods (2, 20, and 60 min). Plasma and whole blood viscosity were measured using a Couette viscometer. Red blood cell morphology was examined by light and scanning electron microscopy. High plasma concentrations of Omegaven increased whole blood viscosity at high and low shear rate (p<0.01 above 20%vol) and plasma viscosity to a lower extent (p<0.05 at 40%vol). Erythrocytes underwent cell swelling (increase of centrifuged hematocrit) and a dose-dependent echinocytic shape transformation, which indicates an intercalation of the compound in the outer hemileaflet of the cell membrane. These effects were already apparent after 2 min of incubation and were fully reversible upon wash-out of Omegaven. We conclude that the intravenous fish oil preparation Omegaven interacts with the outer hemileaflet of the erythrocyte membrane and may affect membrane functions. At pharmacological concentrations these effects seem to be negligeable. PMID- 12652014 TI - An in vitro study of adrenaline effect on human erythrocyte properties in both gender. AB - The possibility that erythrocytes may function as a reservoir for noradrenaline and adrenaline and as a modulator of circulating catecholamine concentrations had been suggested. The aim of this work was to study the adrenaline effect on erythrocyte membrane fluidity, acetylcholinesterase (AChE) enzyme activity, P(50) and erythrocyte deformability and also to verify if the role of adrenaline on erythrocyte properties is sex-dependent. Blood samples from 42 healthy donors were obtained, and its aliquots incubated 30 min without (control) and with 10( 5) M concentrations of adrenaline alone (A(1)) and adrenaline with an alpha and an beta-blocker (A(2)). Results demonstrate that initial AChE values in female are higher (p0.05). We conclude that markers of oxidative stress are not increased and erythrocyte osmotic fragility is not altered in stable COPD patients with normal arterial pH. PMID- 12652017 TI - Changes of vasodilator-stimulated phosphoprotein (VASP) and its phosphorylation in endothelial cells exposed to laminar flow. AB - Vasodilator-stimulated phosphoprotein (VASP), an actin filaments-associated protein expressed mainly in focal adhesions and dynamic membrane regions of endothelial cells (ECs), serves as a substrate for cAMP and cGMP-dependent protein kinases. In this work, we studied the effect of laminar shear stress in vitro on the location and expression of the VASP as well as its phosphorylation associated with actin reorganisation in human umbilical endothelial cells (HUVECs). The distributions of VASP and microfilaments were observed by a fluorescent double staining. The level of VASP expression in cells was quantified by western blot. Experiments showed that, after exposure to a shear stress of 10 dyn/cm(2) for 24 h, besides the elongation and orientation of the cells, and spots of VASP were found along thick stress fibres, particularly at their two extremities. Western blot data showed conversions up to 11.4% and 8.9% of the 46 kD non-phosphorylated form VASP to its 50 kD phosphorylated form after exposures of 1 h and 4 h respectively to the same shear stress. The shear stress of 15 dyn/cm(2) had qualitatively the same effect as that of 10 dyn/cm(2), but with a lower magnitude. However, the shear stress of 2 dyn/cm(2) had much slower and weaker effects on the same tendency. These results suggest that a laminar shear stress can induce VASP translocation and phosphorylation that proceed actin filaments rearrangement along with the flow direction in HUVECs. PMID- 12652018 TI - Three-dimensional model of bone marrow stromal cell culture. AB - In a hematopoietic microenvironment in vivo, spatial organisation of hematopoiesis is possible due to the existence of a three-dimensional framework, the main part of which is formed by a branching population of stromal cells. Most of the previous in vitro studies, concerning long-term bone marrow cultures, were based on a previously prepared, flat adherent layer of stromal cells. There are only few reports concerning the three-dimensional growth pattern of the bone marrow stroma. In the present study we used a new three-dimensional model of the stromal cell culture. The framework for the cultured stromal cells was a structure of a nonliving trabecular bone (Unilab Surgibone). After a period of about four weeks the stromal cells created a spatial network which filled the intertrabecular spaces of the spongy bone. PMID- 12652020 TI - Fretting corrosion behaviour of ball-and-socket joint on dental implants with different prosthodontic alloys. AB - The fretting corrosion of five materials for implant suprastructures (cast titanium, machined-titanium, gold alloy, silver-palladium alloy and chromium nickel alloy), was investigated in vitro, the materials being galvanically coupled to a titanium ball-and-socket-joint with tetrafluoroethylene under mechanical load. Various electrochemical parameters (E(corr), i(corr), Evans diagrams, polarization resistance and Tafel slopes) were analyzed. The microstructure of the different dental materials was observed before and after corrosion processes by optical and electron microscopy. It can be observed that the mechanical load produces an important decrease of the corrosion resistance. The cast and machined titanium had the most passive current density at a given potential and chromium-nickel alloy had the most active critical current density values. The high gold content alloys have excellent resistance corrosion, although this decreases when the gold content is lower in the alloy. The palladium alloy had a low critical current density due to the presence of gallium in this composition but a selective dissolution of copper-rich phases was observed through energy dispersive X-ray analysis. PMID- 12652019 TI - Change in creep behavior of plexiform bone with phosphate ion treatment. AB - The effect of phosphate ions on the mechanical properties of plexiform bone in tension was investigated with an in-vitro model. Bone samples were treated with saline and phosphate ion solutions for three days at 25 degrees C and 37 degrees C and tested in tension. The mechanical properties of the bone samples treated with phosphate were not different than controls (saline treated). Electro kinetic measurements on plexiform bone particles treated with phosphate ions at 37 degrees C showed that phosphate ions alter electro kinetic potentials of bone particles by interacting with bone mineral as compare to saline treated particles near physiological pH. Because of the limited diffusion properties of intact plexiform bone tissue, the tension experiments indicate that, the effect of phosphate ions on the bone mineral-matrix interface is negligible after three days treatment. On the other hand, electro kinetic measurements demonstrated that in a short period of treatment time, phosphate ions diffuse through organic matrix barrier and interact with bone mineral when plexiform bone is in the particle form. As a final experiments bone samples were tested at 37 degrees C in three point bending configuration for three days in saline and phosphate buffer solution. The maximum tension stress generated in bending samples was about 75 percent of the tension yield stress of the samples. The creep experiments showed that the bending rigidity of bone samples tested in phosphate solution reduced in time hence the creep deformation increased compare to control samples tested in saline. This observation is attributed to the acceleration of phosphate ion diffusion into the bending samples due to micro cracks accumulation in bone tissue during the creep experiments which facilitated the phosphate ion interaction with bone mineral. PMID- 12652021 TI - Gamma-irradiation aggravates stress concentration along subsurface grain boundary of ultra-high molecular weight polyethylene (UHMWPE) under sliding fatigue environment. AB - Numerical simulations were carried out using a DEM-based model under a sliding fatigue environment to investigate the influences of differences of mechanical properties between intra-granular portions and inter-granular portions on local stress-strain fields and to determine the effect of gamma-irradiative degradation of UHMWPE components on delamination. The predicted stress fields in the gamma irradiated UHMWPE component were highly nonuniform and were quite different from those predicted in the non-irradiated UHMWPE component. The stress distributions were intensely affected by the structural inhomogeneousness of networked grain boundaries. Marked stress concentrations were observed along subsurface grain boundaries in the gamma-irradiated UHMWPE. The gamma-irradiated UHMWPE showed a steep increase in the maximum equivalent stresses for an increase in the number of sliding. As the number of sliding increased, moreover, the region where the maximum equivalent stress was observed, moved deep into the specimen. These findings suggest that fatigue damage with plastic deformation proceeds mainly at a subsurface region in the gamma-irradiated UHMWPE and at a superficial region in the non-irradiated one. Therefore, gamma-irradiation is an important factor in accelerating fatigue crack initiation resulting in delamination. PMID- 12652022 TI - Microscopic destruction of ultra-high molecular weight polyethylene (UHMWPE) under uniaxial tension. AB - To examine the effects of a networked substructure of granular agglomerate on a style of destruction in prosthetic ultra-high molecular weight polyethylene (UHMWPE), uniaxial tensile simulations were carried out using the numerical model based on the discrete element method (DEM). The numerical simulations were performed taking the difference of mechanical characteristics between inter granular and intra-granular portions of UHMWPE into consideration. A significant increase in stress and strain was observed along grain boundaries where micro cracks were initiated. This finding suggests that the large difference of mechanical properties between intra-granular and inter-granular portions causes significantly increased local stresses and strains in the vicinity of grain boundaries. The tensile simulation resulted in intra-granular destruction, which had good agreement with a result of previous experimental observation. This is presumably because the directions of principal shear stresses do not coincide with those at stress-concentrated grain boundaries. The dependence of the style of material destruction on forms of loading application could be explained by the relationship between directions of principal shear stress and reorientations of stress-concentrated grain boundaries. PMID- 12652023 TI - Artificial vitreous replacements. AB - Vitreoretinal pathologies remain the significant leading cause of blindness, after cataract, throughout the world. Biomaterials as a whole, and vitreous substitutes in particular, have had a major role to play in vitreoretinal surgery. History has witnessed the advent of vitreous substitutes from collagen and hyaluronic acid to gases, silicone oils and perfluorocarbon liquids and now to polymer hydrogels. Such a long list only underscores the fact that we have been found wanting in our quest for an ideal vitreous substitute. This review focuses on the merits and demerits of various vitreous substitutes, which have been tried to date, clinical as well as experimental. The properties required of an ideal substitute and the performance of the vitreous substitutes with respect to these criteria have been described. Finally, the concluding note dwells on the future directions for biomaterial research in the posterior segment of the eye and undermines the role of tissue engineering in vitreoretinal regeneration. PMID- 12652024 TI - Finite element analysis of a model of a therapeutic shoe: effect of material selection for the outsole. AB - The finite element analysis method was used to perform a sensitivity study of the effect of the materials used for the two layers of the outsole (high-density polyethylene, HDPE, and polyurethane, PU) of a "solid rocker-bottom" design of a therapeutic shoe on the responses (stresses and displacements) of a model of the shoe. It was found that the aforementioned materials choice affected the model responses in a noticeable manner. For example, when HDPE was used for the top layer of the outsole, the von Mises stress at the interface between the bottom of the foot and the top layer of the insole ranged from being about 8% lower to being about 62% higher compared to when PU was used. The implications of this finding are discussed. PMID- 12652025 TI - Preparation and osteocompatibility of hydroxyapatite coated on titanium from the reaction of sputtered CaO and vaporized P2O5. AB - Hydroxyapatite (HA) and other calcium phosphates were synthesized on titanium plates by a solid-gas state reaction of sputtered CaO and vaporized P(2)O(5). The calcium phosphates formed were HA, beta-tricalcium phosphate (beta-TCP; Ca(3)(PO(4))(2)), beta-calcium pyrophosphate (beta-PYR; Ca(2)P(2)O(7)), and beta calcium metaphosphate (beta-MET; Ca(2)(PO(3))(2)). Their formation depended on the ratio of the sputtered CaO and the reacting P(2)O(5). For a mole ratio of CaO/P(2)O(5)=4 (Ca/P=2), an HA film was synthesized. The surface roughness increased by over seven times after the solid-gas state reaction from Ra = 0.16+/ 0.02 microm (for the CaO film) to Ra = 1.15+/-0.25 microm (for the reacted film). The synthesized HA film-coated titanium plates and control non-coated titanium plates were implanted in the femora of two dogs for a period of two, four and 12 weeks, and observed using a soft X-ray radiograph and histological sections. New bone formation was observed without any connective tissue at four weeks around the HA film, whereas over the 12 week experimental period, there was no new bone formation around the control and connective tissue was observed over all periods, reaching a thickness of more than 200 microm at 12 weeks. PMID- 12652026 TI - Modeling of biomedical interfaces with nonlinear friction properties. AB - Proper isotropic and anisotropic friction constitutive equations are developed based on previous friction measurements at cancellous bone-porous coated implant interfaces exhibiting nonlinear load-displacement curves. The simulated friction response is dependent on relative tangential displacements in both orthogonal directions. The interface constitutive matrix contains cross-stiffness terms identical in isotropic friction but different in anisotropic friction. These terms are due mainly to nonlinearity in response and vanish in unidirectional friction along a principal direction and in cases with Coulomb or linear friction. The interface ultimate resistance is evaluated by an elliptic criterion which becomes circular in isotropic cases. These constitutive relations are implemented in a finite element program which is employed to analyze a bone cube sliding on top of a porous-surfaced metallic plate, an experimental model used in our earlier measurements. The results for both isotropic and anisotropic frictions demonstrate the coupling between two orthogonal directions. The direction of resultant displacement under a variable load coincides with that of the load only when the friction is isotropic with coupling terms considered. In anisotropic friction, the resultant displacement occurs in a direction different from that of loading. Our previous bi-directional measurements corroborate well the findings of this study. PMID- 12652027 TI - In vitro cytocompatibility studies of Diamond Like Carbon coatings on titanium. AB - Diamond like carbon (DLC) films were deposited on to titanium (Ti) substrates by Plasma Enhanced Chemical Vapour Deposition (PECVD) process. The quality of the films were checked by Raman spectra and nano-hardness tests. The cytocompatibility of titanium and DLC coated titanium were studied using continuous cell lines of mouse fibroblast cells ( L-929), Human Osteoblast cells (HOS) and primary human umbilical cord vein endothelial cells (HUVEC). The cellular responses to the materials were assessed both quantitatively and qualitatively. The adhesion and spreading of cells on materials were compared using Ti as a control. Present study indicates an improved cytocompatibility of DLC coated Ti in comparison to bare Ti. PMID- 12652028 TI - A new porous titanium-nickel alloy: part 2. Sensitization, irritation and acute systemic toxicity evaluation. AB - Porous titanium-nickel (PTN) represents a new biomaterial with orthopedic applications as a long-term implant. Because of its nickel content, PTN was tested for its potential to stimulate sensitization, irritation, and systemic toxicity reactions after semi-physiological extraction. In order to do so, an in vivo biocompatibility evaluation was performed following three ISO-standardized methods using accepted animal models for immunity testing: the classical skin sensitization assay (Buehler patch test) in guinea pigs, the rabbit intracutaneous test, and the systemic injection test in mice. The Buehler patch test in guinea pigs revealed no significant change in skin reactions such as erythema or swelling between the induction and the challenge period. In the rabbit intracutaneous irritation test, no irritation or sensitization reactions were observed in saline-extracted PTN samples. Negligible to slight irritation was observed at some of the sites involving PTN samples extracted in cottonseed oil, however the resulting primary irritation index was similar to the one elicited by the blank solution itself. Finally, no toxic symptoms were observed with any of the mice injected with porous titanium-nickel extracts during the acute systemic toxicity test. Based on the above results, porous titanium-nickel is considered to be a non-sensitizing, non-irritant, and non-toxic biomaterial for medical applications. PMID- 12652029 TI - Tensile adhesion of type I collagen to titanium alloy and calcium phosphate coated surfaces with different roughness values. AB - The purpose of total joint arthroplasty is to reduce pain and restore function. Its success depends on the formation of a new bone that stabilizes the prosthesis. The proposed solution for this important problem is to have bio coated implant surfaces which are more conductive to bone growth. Additionally, collagen has long been used as a matrix for medical applications, because of its biocompatibility and adaptability. In this study, a test method for measuring the tensile adhesion strength of collagen to titanium alloy and calcium phosphate coated surfaces with different roughness values was developed, in order to evaluate how well the collagen adheres to the metallic and bio-coated surfaces. A precision motion system was used to stretch gels that were adherent to the plate surfaces. The tests were done in DMEM solution. The adhesive strength between the collagen gel and plate was significantly higher for calcium phosphate coated surfaces. Adhesive strength was highest in the sample with the highest roughness value. PMID- 12652030 TI - Reinforcement of acrylic resins for provisional fixed restorations. Part III: effects of addition of titania and zirconia mixtures on some mechanical and physical properties. AB - Acrylic resins have been used in many different applications in dentistry, especially in the fabrication of provisional fixed partial dentures. Ideally, a provisional crown and bridge material should be easy to handle and should protect teeth against physical, chemical, and thermal injuries. Some of the problems associated with this use are related to the material's poor mechanical properties. It has been demonstrated that acrylic resin can be strengthened through the addition of structural component of different size distributed in the acrylic matrix, thus forming a composite structure. The purpose of this study was to investigate the addition effects of mixtures of titania (titanium dioxide, TiO(2)) powder and zirconia (zirconium dioxide, ZrO(2)) powder being incorporated with pre-polymerized beads mixed in monomer liquid, on some mechanical and physical properties of PMMA resin. The pre-polymerized powder poly(methyl methacrylate) resin was admixed with titania and zirconia powder. A mixing ratio was controlled by volume % of 0, 1.0, 2.0, and 3.0 (samples with 0 v/o served as control groups). For using mixture of titania and zirconia, total amount of the mixture was controlled by volume % of 1.0, 2.0, and 3.0, in which titania and zirconia were mixed at the ratio 1 :1, 1 :2 and 2 :1. Prior to mechanical tests, all rectangular-shaped samples (25 mm x 2 mm x 5 mm) were stored in 37 degrees C distilled water for 7 days after polishing all six sides of samples. Samples were then subjected to the three-point bending flexion test to evaluate the bending strength as well as the modulus of elasticity. Weight gain and exothermic reaction survey were investigated as well. All data were collected and analyzed with one-way analysis of variance (ANOVA) and Sidak method (p=0.05). It was found that the addition of particles generally decreased the water absorbed by the composite system. Only 1 percent by volume concentration of 1 :1 ratio and 2 percent by volume concentration of 1 :2 and 2 :1 ratios had significantly higher strength than control group. There was significantly higher toughness (in terms of energy-to-break) for 1 percent by volume concentration of 1 :1 ratio and 2 percent by volume concentration of 2 :1 ratio than control group. There was no significant difference between control group and all percent by volume combinations in modulus of elasticity. In addition, there was no significant variations of exothermic reaction. PMID- 12652031 TI - Biomechanical analysis of the lumbar spine with anterior interbody fusion on the different locations of the bone grafts. AB - The anterior lumbar interbody fusion is the common procedure in the management of the degenerated disc in the lumbar spine, but the biomechanical behavior of the fused segment would be changed because of the implantation of bone graft at the different locations. To investigate the biomechanical alteration, the study applied the finite element model to undergo the stress analysis.A three dimensional finite element model of the lumbar spine was established, and modified to the three fusion models consisted of the bone graft at the anterior site, the middle site and the posterior site, respectively. The 12 N m flexion and the 10 N m torsion with pre-load 150 N were imposed on the L1 vertebral body. The results of the finite element model indicated that placing bone graft at anterior site could effectively resist flexion moment, and decreased the tensile force of the posterior ligaments about 15% above. Placing bone graft at posterior site could resist torsional moment, and also led to none of contact force of the facet joint in the fused segment. However, wherever the bone grafts were placed, stress slightly increased on the disc adjacent to interbody fusion about 5% below. PMID- 12652032 TI - Repassivation of a high chromium stainless steel orthopaedic alloy. AB - The repassivation of Ortron90, a high chromium stainless steel alloy, was studied to assess the effect of passive layer removal on corrosion levels. Cathodic dissolution of the passive layer followed by potentiostatic experiments within the passive region were employed to study the repassivation process. The scratch method, carried out for comparison purposes, yielded current densities similar to those obtained by cathodic dissolution. The detailed analysis of the slopes of plots of log I/log t indicated that the repassivation did not follow any of the existing growth models. It also suggested that the slope depended on the extent of repassivation and reached a minimum when three to five monolayers of oxide were completed. The virtual potentiodynamic plots suggested the presence of some form of protective layer within 5-10 seconds and the change in open circuit potential vs time indicated that the passive layer present after 10 seconds of repassivation continued to evolve for about 1000 to 1500 seconds possibly representing a transition between two different forms such as oxide and a oxyhydroxides. Estimation of the effect of localized removal of passive layer on the corrosion indicated a dramatic increase in local corrosion density. These results show that a fast repassivation rate is an important requirement for biocompatibility of alloys used to manufacture dental and orthopaedic devices. PMID- 12652034 TI - Nerve cuff electrode with shape memory alloy armature: design and fabrication. AB - In this paper, a new nerve cuff electrode with shape memory alloy armature is presented. The proposed electrode is dedicated either to peripheral nerve stimulation or recording and its manufacturing does not require any expensive or complex technique. Shape Memory Alloy (SMA) armature ensures the complete and firm closing of the electrode, so that the complexity of the installation procedure is considerably reduced. A preliminary analysis of the electrode mechanical behavior prior, during and after installation has been done through numerical simulations and in vitro testing. It was proved theoretically and experimentally that the SMA electrode closes completely with an appropriate few second delay after its installation. No external fixation such as sutures is needed to secure permanent electrode-nerve contact. Furthermore, theoretical analysis has shown that the design of SMA electrode can be adapted for safe close fitting installation, thanks to the device partial opening in case of nerve swelling. PMID- 12652033 TI - Screening of stabilized crosslinked polyethylene using a novel wear tester. AB - A novel pin-on-disk type wear tester is described allowing a rapid screening of different types of polyethylene under both unidirectional and multidirectional sliding motion. The wear of four polyethylene materials sliding against a roughened CoCrMo alloy was evaluated: a non-irradiated UHMWPE, a UHMWPE irradiated with a dose of 25 kGy in air, and two types of crosslinked UHMWPE (100 kGy, air), which were subjected to a stabilization heat treatment in nitrogen at 155 degrees C for 72 hours (XLPE I) and in water at 130 degrees C for 72 hours (XLPE II), respectively.Under multidirectional sliding conditions both types of XLPE exhibited significantly less wear with respect to the 25 kGy irradiated UHMWPE and the non-irradiated UHMWPE, even under the rough counterface conditions applied. Under unidirectional sliding motion both types of XLPE exhibited the highest wear of all materials tested, because the orientation hardening effect acting under linear lubricated condition is less pronounced for crosslinked polyethylene. PMID- 12652035 TI - Influence of Charnley hip neck-angle inclination on the stresses at stem/cement and bone/cement interfaces. AB - The present work aims to elucidate the influence of neck-stem angle inclination on the principal normal and shear stress distributions and values along the interfaces of the stem/cement and bone/cement in a cemented Charnley curved back femoral component. The same stresses were also examined for the intact bone that was considered as a reference for the obtained results. The interface is considered the weakest link in the structure and its endurance limit to failure is much less than the limit of the adjacent materials. Interface loading is by far the influential aspect governing the induced interface stresses in femoral total hip replacement. Therefore, higher values and nonuniform distribution of stresses at the interface may lead to loosening and crack initiation and propagation that usually precede the stem fracture. PMID- 12652036 TI - Composite resorbable polymer/hydroxylapatite composite screws for fixation of osteochondral osteotomies. AB - The aim of this study was to evaluate biomechanically the healing of an osteochondral fragment created in the distal sheep femur in response to fixation with a resorbable composite screw made of polylactide and hydroxylapatite. Pure poly(L-lactide) screws were used for comparison. At follow-up times of 4 or 8 weeks, specimens were examined with standard radiography, biomechanics, and histology. The intact contralateral femur served as a control. Only minimal signs of polymer degradation were seen in the histologic specimens. At 8 weeks, most osteotomies had healed completely and there was no difference in compressive strength and elastic modulus of cylindrical cores between the two types of biodegradable implants used. The width of the repair tissue at the tissue-implant interface was 250+/-50 micro m representing a clear transition zone of newly formed trabecular bone separating the implant from the surrounding plexiform bone. We conclude that relatively large polylactide implants, blended with hydroxyapatite, are capable of fixing an osteochondral fragment in an animal model. Biomechanical data assessing the quality of the bone formed at the osteotomy sites were found to be equivalent when compared to the control poly(L lactide) implants of similar design and size. In addition, hydroxylapatite composite implants showed benign tissue responses and good implant osteointegration. Results suggest that hydroxylapatite composite screw implants can be used for similar indications as pure poly(L-lactide) implants in current clinical use. PMID- 12652037 TI - Deep brain stimulation for Parkinson's disease. A critical re-evaluation of STN versus GPi DBS. AB - Deep brain stimulation (DBS) in the subthalamic nucleus (STN) and the internal segment of the globus pallidus (GPi) is increasingly being used for the treatment of advanced Parkinson's disease (PD). Although both targets have demonstrated clinical efficacy in the treatment of the cardinal motor signs of PD, the STN has gained greater popularity and is now considered the site of choice by most centers performing these procedures. This preference stems predominately from the belief that STN DBS provides greater improvement in reducing the motor manifestations of PD and allows a reduction in dopaminergic medication not permitted with GPi DBS. There are, however, a number of issues that must be considered before abandoning GPi in favor of STN as the surgical target of choice for DBS. The maximal benefit reported for GPi stimulation is not significantly different than that reported for the STN, 67 versus 71%, and while reductions in medication are required with STN stimulation to avoid inducing dyskinesia, GPi stimulation may directly suppress dyskinesia obviating any need to reduce medication. As such, many centers may not attempt to reduce antiparkinsonian medication with GPi DBS. In addition, there are significantly more reports of changes in mood, behavior and a higher incidence of adverse events reported for STN stimulation. Most studies of DBS are nonrandomized, assessment protocols are not standardized, and lead locations are not reported. Thus, before drawing conclusions regarding the optimal site for DBS for advanced PD we must take a critical eye to the present data and address the outstanding questions that remain with well-designed clinical trials that evaluate motor, nonmotor and adverse events and address the above clinical variables by randomizing patients, using standardized methods of assessment and defining the lead location. PMID- 12652038 TI - MRI-directed subthalamic nucleus surgery for Parkinson's disease. AB - The subthalamic nucleus (STN) is now regarded as the optimal surgical target for the treatment of medically refractory idiopathic Parkinson's disease. In our center, a predominantly MRI-directed method has been developed for targeting the STN. The STN is localized on T2-weighted images from a 1.5-T MRI scanner. Long acquisition, high-resolution images are acquired in both the axial and coronal planes under strict stereotactic conditions with the patient under general anesthesia. The boundary of STN is co-registered in both planes to give optimal 3 dimensional target definition. Stereotactic coordinates of the dorsolateral STN are recorded and the trajectory is planned down the axis of the nucleus in the coronal plane. Initially, per-operative macrostimulation was used for adjustment at the target prior to unilateral subthalamotomy in 26 patients. Five patients were lost to follow-up. Assessments of the lesions in post-operative images confirmed successful localisation of the lesions within the dorsolateral STN in all of the remaining 21 cases. In a subsequent series of 19 patients treated by deep brain stimulation (DBS), unilateral in 1 patient and bilateral in 18, the STN was targeted using the same MRI-directed method, guide tubes and radio-opaque stylettes were implanted, and target verification was entirely MRI-based. Following implantation of the guide tubes and stylettes, assessments of the per operative MRI images for the 37 STN targetings confirmed a mean target error, between the stylette and the desired target in the axial plane, of 0.3 mm mediolaterally (SD = 0.4) and 0.4 mm anteroposteriorly (SD = 0.4), with median errors of 0.5 mm. This study demonstrates that MRI-directed targeting of the STN through guide tubes is accurate, and allows direct verification and corrections as necessary. Cumulative frequencies predict that the majority of DBS electrodes placed in this manner will be within 0.5 mm of the planned target. Because physiological methods are not required, the whole procedure can be performed under general anesthesia. We feel that planning with reference to a standard atlas is unreliable and not significantly helped by the addition of microelectrode recording, the accuracy of which in the axial plane is dependent upon the distance between the recording trajectories, which is typically 2 mm. PMID- 12652039 TI - Safety and risk of microelectrode recording in surgery for movement disorders. AB - There is an ongoing controversy about whether it is necessary to use microelectrode recording (MER) techniques in stereotactic surgery for Parkinson's disease and other movement disorders. This paper consists of a critical review of the published literature in order to analyze the value of MER in providing safe, efficient and accurate functional stereotactic surgery. Review of the literature revealed that MER techniques do not necessarily improve targeting accuracy or clinical results, compared to techniques using impedance monitoring and macrostimulation. In terms of safety for the patients, however, MER techniques are relatively safe, but non-Mer techniques, based on macrostimulation-guided surgery, are at least five times safer. PMID- 12652040 TI - Exploration of the role of the upper brainstem in motor control. AB - The rostral areas of the brainstem have been extensively studied in higher mammals and to a lesser extent in humans in the last two decades, looking for anatomical, electrophysiological and neurochemical evidence of involvement in the initiation and control of voluntary movement. This has come with the realisation that the axial symptoms of advanced Parkinson's disease (PD), like akinesia, postural impairment and gait freezing, are relatively less responsive to current medical and surgical treatments directed primarily at the basal ganglia and thalamus. The pedunculopontine nucleus (PPN) is one such area of interest. We have found that lesioning and electrical stimulation at high frequencies of the PPN region in the normal behaving primate induces akinesia, and low frequency stimulation can induce tremor. Micro-injections of gamma-aminobutyric acid (GABA) receptor A agonist, muscimol, into the PPN decreases activity. In the 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP) treated Parkinsonian primate model, bicuculline, a GABA(A) antagonist, can alleviate akinesia when infused into the PPN region. This may suggest new targets for treating the intractable akinetic symptoms of advanced PD. PMID- 12652041 TI - Deep brain stimulation for dystonia in adults. Overview and developments. AB - The renaissance of functional neurosurgery in the treatment of Parkinson's disease has sparked also the interest in other movement disorders which are refractory to medical treatment. Deep brain stimulation (DBS) has been used only since a few years in dystonia patients. This review summarizes the available data on pallidal and thalamic DBS for various dystonic syndromes. The major advantage of DBS as compared to radiofrequency lesioning is that it allows performing contemporaneous bilateral surgery with relatively low morbidity in these patients. The posteroventral lateral globus pallidus internus (GPi) has been the preferred target in most instances, thus far. While phasic dystonic movements may improve early after surgery, the response of tonic dystonic movements to chronic stimulation may be delayed. The most beneficial results have been achieved in patients with primary genetic generalized and segmental dystonia, myoclonic dystonia, and complex cervical dystonia. Outcome has been varied in patients with other dystonic disorders, in particular those with secondary dystonia. Most studies have reported on relatively short follow-up periods, on single cases, or were retrospective. Pallidal DBS has been shown to be effective in complex cervical dystonia yielding both symptomatic and functional benefit for up to 2.5 years of follow-up. Dramatic improvement has been obtained in children and in adults with DYT1 positive dystonia. Also, patients with non DYT1 genetic dystonia achieved sustained benefit for up to 2 years of follow-up. Preliminary experience indicates that choreoathetosis in patients with cerebral palsy responds less well to pallidal DBS, and that it may not be effective at all in some patients. In single instances unilateral pallidal DBS has been shown to yield valuable benefit in patients with hemidystonia. The experience with DBS for treatment of Meige syndrome and other focal dystonias has been explored only recently. There is much less experience with thalamic DBS for dystonia. Thalamic DBS has been shown to be effective in single cases with posttraumatic dystonia, postanoxic dystonia and paroxysmal nonkinesigenic dystonia. Future perspectives of DBS for treatment of dystonia include the development of new technology, the evaluation of the possible role of other targets, and carefully planned studies to further establish the role of surgery. PMID- 12652042 TI - Deep brain stimulation for dystonia. Surgical technique. AB - Stimulation electrodes are implanted under general anesthesia, without intra operative electrophysiology or clinical testing, based only on stereotactic MRI and direct anatomical localization of the postero-ventro-basal GPi. We retrospectively analyzed the surgical procedure that has been designed and implemented in our center, using the Leksell G frame, for initiating deep brain stimulation in 65 dystonic patients. We report the surgical technique and the hardware and software complications. We recommend immediate postoperative stereotactic MRI under general anesthesia as a prerequisite to check the reliability of MR acquisition (magnet stability) and the exact localization of each electrode. This technique allowed us to reduce the duration of the operation to 4 h, including general anesthesia, frame fixation, MRI acquisition, implantation of two electrodes under radioscopic control, immediate postoperative stereotactic MRI and frame removal. Surgery-related morbidity was very low with a 0% hemorrhage rate and three delayed unilateral infections re-operated 6 months later. Hardware and software complications were rare. The advances in 3D-MR imaging permit the electrode implantation for deep brain stimulation without resorting to intraoperative localization techniques, which is especially helpful in children and for treating dystonia. The maximum follow-up period is 58 months (first case: November 1996). GPi stimulation has proven to be an effective treatment for most dystonic syndromes with particular efficacy in the disease due to the DYT1 mutation. PMID- 12652043 TI - Transcranial magnetic cortical stimulation relieves central pain. AB - Extradural cortical stimulation for neurogenic pain is a recent addition to the field of functional neurosurgery. About 50% of patients with central pain draw benefit in the long run. However, there is an urgent need for prognostic factors in order to cut the costs of the procedure. In this paper we report a statistically significant correlation between the subhypnotic propofol test, transcranial magnetic cortical stimulation (TMS) and the actual short-term outcome of extradural cortical stimulation in 9 patients. The propofol test and TMS appear to predict short-term effects of extradural cortical stimulation. PMID- 12652045 TI - Systematic review of studies comparing the anti-caries efficacy of children's toothpaste containing 600 ppm of fluoride or less with high fluoride toothpastes of 1,000 ppm or above. AB - OBJECTIVE: To determine the clinical effectiveness of low fluoride (F) toothpastes, containing 600 ppm or less, with toothpaste containing 1,000 ppm or more fluoride in preventing dental caries. DESIGN: Systematic review of randomised controlled trials comparing low fluoride toothpastes containing 600 ppm F or less with toothpastes containing 1,000 ppm or more in children or adults. RESULTS: Seven randomised controlled trials were included. These were categorised into two groups depending on the fluoride concentration in the low fluoride group (250 and 500 ppm) and analysed separately. 250 PPM: Results of the meta-analysis were statistically significant (p = 0.002 and 0.0005) and in favour of the control group (1,000 ppm). DFS increments in the 250-ppm group were 0.6 0.7 greater than the 1,000-ppm group. 500 PPM: Only two studies were in this category, and one of them failed to present the baseline caries levels, so a meta analysis was not carried out. CONCLUSION: 250 ppm fluoride dentifrice was not as effective in caries prevention in permanent dentition as dentifrice containing 1,000 ppm F or more. More studies have to be carried out to test the anticaries efficacy of 500 against 1,000 ppm and above. PMID- 12652044 TI - Dental caries related to plasma IgG and alpha1-acid glycoprotein. AB - This study was aimed at determining whether dental caries is associated with induction of the systemic immune system or cytokine response. For this purpose, 85 children from Den Pasar, Bali, Indonesia, aged 6-7 years, were examined clinically and blood plasma was obtained via finger puncture. The concentrations of the acute-phase protein alpha(1)-acid glycoprotein (AGP), total IgG and the specific IgG and IgM immunoglobulins against Streptococcus mutans were determined. Immunoelectrophoresis was used for the determination of the AGP concentration and ELISA for IgG and IgM detection. The mean dmft of the whole group was 8.8 +/- 2.9, the mean number of infected pulps was 3.9 +/- 2.2 and the mean number of abscesses was 0.5 +/- 0.8. The plasma concentration of AGP ranged between 0.13 and 1.6 mg/ml serum (mean 0.86 +/- 0.26 mg/ml). Stepwise regression analysis revealed that the concentration of IgG against S. mutans (log transformed) was significantly correlated with dmft (adjusted r(2) = 0.083, standardized beta coefficient = 0.31, p = 0.008). When the concentration AGP was included in the model the correlation improved significantly (for IgG: adjusted r(2) = 0.157, standardised beta coefficient = 0.36, p = 0.002; for AGP: beta coefficient = -0.30, p = 0.009). The results suggest a relationship between caries and systemic parameters of inflammation. On the basis of this, severe caries might have consequences on the general health of the subject. PMID- 12652046 TI - Susceptibility of enamel surfaces to demineralization after application of fluoridated carbamide peroxide gels. AB - The aim of the present study was to evaluate the effect of experimental, fluoridated carbamide peroxide gels on formation of erosively induced demineralization of enamel. Seventy-five bovine enamel specimens were polished for microhardness determination and evenly distributed among 5 groups (A-E). The specimens were treated with 10% carbamide peroxide gel (8 h) and subjected twice to remineralization for 2 h in artificial saliva and demineralisation for 90 s in 1% citric acid, pH 2.2. The cycles of treatment with carbamide peroxide and twofold re- and demineralization were repeated three times. The carbamide peroxide gels were different in pH and fluoride content. Gel A (pH 7.0) and gel B (pH 5.7) were fluoridated (0.5% F), gel C (pH 7.0) and gel D (pH 5.7) were not fluoridated. In control group E the samples were not treated with a gel, but stored in 100% humidity for 8 h instead. Knoop microhardness of the specimens was assessed directly after polishing, and after each carbamide peroxide treatment and demineralization. All specimens showed a loss of microhardness at the end of the experiment. After 3 days, the controls revealed a significantly lower hardness loss compared to the specimens treated with the carbamide peroxide gels. Surface softening was significantly lower for the specimens of group A compared to the remaining groups (B-D), which were not significantly different among each other. It is concluded that treatment with either fluoridated or unfluoridated carbamide peroxide gels, at either neutral and acidic pH, renders enamel more susceptible to demineralization. Use of a fluoridated neutral gel decreases the degree of surface softening compared to the other gels investigated. PMID- 12652047 TI - Influence of olive oil emulsions on dentin demineralization in vitro. AB - The effect of two different concentrations of olive oil emulsions on development of artificial caries-like dentin lesions under severe demineralizing conditions was investigated. Bovine dentin samples (n = 180) were ground flat, polished, divided into four groups, and subjected to three demineralization cycles per day. Samples were stored in one of the following solutions for 5 min prior to demineralization in a buffer solution (pH 5): Group 1: 50% oil emulsion (olive oil and distilled water); group 2: 5% oil emulsion; group 3: distilled water; and group 4: 1,500 ppm sodium fluoride. Daily up to 9 days, lesion depth (ld) and mineral loss (deltaZ) were determined by means of microradiography and analyzed by ANOVA and Tukey's studentized range test (p < or = 0.05). Lesion depth increased with time for all groups. Mineral loss increased in groups 1-3. A small but significant decrease in mineral loss was observed following treatment with lipid emulsions as compared to treatment with distilled water, but fluoride treatment was considerably more effective. Mean mineral loss (means +/- SD in vol% x microm) averaged over the study period was 4,368 +/- 1,599, 4,536 +/- 1,823, 4,849 +/- 1,798, and 789 +/- 177 for group 1, 2, 3, and 4, respectively. Ratio (deltaZ/ld) remained constant around 30 vol% for groups 1-3, but decreased for group 4. In conclusion, externally provided lipids have the potential to reduce dentin demineralization in vitro. PMID- 12652048 TI - Inhibitory effects on selected oral bacteria of antibacterial agents incorporated in a glass ionomer cement. AB - The objectives of the study were to investigate the antimicrobial efficacy, over time, of combining antibacterial agents with a glass ionomer cement (GIC). This was assessed using an agar diffusion test. Chlorhexidine hydrochloride, cetylpyridinium chloride, cetrimide and benzalkonium chloride were added to Fuji IX GIC at 0, 1, 2 and 4% w/w. Antibacterial-GIC specimens were placed onto agar plates inoculated with one of six bacterial species (Streptococcis, Lactobacillus, and Actinomyces, two each) and the area of inhibition calculated after 24 h incubation. The experiment was repeated weekly and at week 11 the surface of the specimen was abraded prior to replacing on inoculated agar plates. Control specimens of the GIC produced no bacterial inhibition. The antibacterial GIC combination specimens showed significant inhibition which decreased at different rates over the test period. Resurfacing of the specimens showed a dramatic increase of antibacterial action similar to levels produced on week 1. CT-GIC showed the greatest (p < 0.005) inhibitory effect throughout the experimental period for 4 out of 6 test bacteria. The addition of antibacterial agents to Fuji IX creates a GIC material with significant antimicrobial action in vitro which is dependent on concentration and type of antibacterial agent, and appears to be associated primarily with a release of the antibacterial from the surface layer of the specimen. PMID- 12652049 TI - Diagnostic outcome of methods frequently used for caries validation. A comparison of clinical examination, radiography and histology following hemisectioning and serial tooth sectioning. AB - The aim of this study was to compare the caries diagnostic outcome of four methods frequently used as validation for dental caries. The diagnostic outcome of clinical examination (CL), radiography (RA), and histology after serial tooth sectioning (HI-serial) on 373 approximal and 158 occlusal surfaces was compared, and furthermore histology after hemi- (HI-hemi) and serial sectioning on another 113 approximal and 53 occlusal surfaces was compared. Two thresholds for each method (CL: (1) sound vs. all caries scores, and (2) non-cavitated vs. cavitated lesions; RA and HI: (1) sound vs. all caries scores, and (2) no dentine vs. dentine lesions) were evaluated. In general, large differences in diagnostic outcomes were observed with the various methods. At threshold 1, CL resulted in significantly more lesions than both RA and HI-serial on approximal surfaces, and than RA on occlusal surfaces. At threshold 2, no significant differences between CL, RA and HI-serial were found on approximal surfaces, but on occlusal surfaces significantly more lesions were diagnosed with RA and HI-serial than with CL. Significantly more occlusal lesions were found by HI-serial than by RA at both thresholds 1 and 2. On approximal surfaces, a similar result was found only at threshold 1. On approximal surfaces, significantly more lesions were diagnosed with HI-serial than with HI-hemi at both thresholds 1 and 2. On occlusal surfaces the same was found only at threshold 1. The intra-observer reproducibility was higher using HI-serial than using RA and CL. PMID- 12652050 TI - Relationships between a clinical-visual scoring system and two histological techniques: a laboratory study on occlusal and approximal carious lesions. AB - One aim of the present laboratory study was to determine whether a visual scoring system (ERK) developed for occlusal caries could be applied to approximal lesions. A new histological technique (autofluorescence, AF) recognises dentine that is soft and would be removed with an excavator during operative treatment. A second aim was to investigate the relationship between the visual scoring system (ERK) and AF of dentine both occlusally and approximally. The sample comprised 93 extracted teeth chosen to represent the range of visual scores on approximal and occlusal surfaces. After sectioning through the investigation site, the cut faces were examined in a stereomicroscope and the depth of demineralization was scored. Autofluorescence was viewed with a confocal laser scanning microscope. Results showed reasonable correlation between the visual scores and the stereomicroscope histological evaluations for occlusal surfaces and non-cavitated approximal surfaces. However, cavitated approximal surface lesions were less advanced histologically than cavitated occlusal carious lesions. The AF technique indicated that several lesions with intact surfaces would have had soft, excavatable dentine, whereas several with microcavities would not. PMID- 12652051 TI - Influence of different variables on the inter-municipality variation in caries experience in Danish adolescents. AB - This investigation sought to estimate the influence a number of variables had on the inter-municipality variation in caries experience across Denmark. Unit of measurement was the municipality with public clinics. Mean DMF-S and %DMF-S = 0 of 15- and 18-year-olds in 1999 were obtained from 204 and 143 municipalities, respectively, out of a total number of 206 municipalities with public clinics. The independent variables were: cost per child per year; children/dentist ratio; auxiliary personal/dentist ratio; fluoride concentration in the water supply [F]; average personal income; % of mothers of the 15- and 18-year-olds with < or =10 years education (EDU-15 or EDU-18); proportion of immigrants; and size of the municipality. Multiple regression analyses disclosed that [F] (p < 0.001) and EDU 15 (p < 0.001) were significant variables among the 15-year-olds explaining 45% of the variation in mean DMF-S and 31% of the variation in % DMF-S = 0. With respect to the 18-year-olds, [F] (p < 0.001) and average personal income (p < 0.001) explained 53% of the variation in mean DMF-S and 30% of the variation in %DMF-S = 0. Few municipalities were characterized as outliers with significantly lower or higher observed caries experience than expected. It is concluded that there is room for other explanatory factors--first and foremost the professional effort made in the individual Public Dental Health Service to control caries. PMID- 12652052 TI - Metabolic control as a modifier of the association between salivary factors and dental caries among diabetic patients. AB - The aim here was to analyze the role of metabolic control as a modifier of the association of salivary factors with dental caries among diabetic patients. Cross sectional data were gathered from 149 insulin-dependent diabetes mellitus patients by means of clinical examination, a quantitative questionnaire and patient records. Dental caries was measured as the number of decayed surfaces. The data were analyzed using negative binomial regression models. HbA1(c) level appeared not to be associated with dental caries. Of the salivary factors, only high levels of mutans streptococci (MS) and lactobacilli were associated with dental caries. Among the subjects with HbA1(c) > or =8.5, the presence of dental caries was more distinctly associated with higher levels of MS (RR = 3.86, CI: 1.39-11.96) and lactobacilli (RR = 3.59, CI: 1.44-9.33) compared to those with HbA1(c)<8.5. It can be concluded that poor glycemic control strengthens the positive association of MS and lactobacilli with dental caries. PMID- 12652053 TI - Chromosomal insertions and deletions in Streptococcus mutans. AB - Many isolates of Streptococcus mutans lack the ability to ferment melibiose and other sugars. We previously reported that this was commonly due to a chromosomal deletion and, in the present study, sequence information from the S. mutans genome project was used to design PCR primers to explore the nature and extent of the deletion. In all melibiose-negative strains examined, there was an incomplete insertion element, ISSmu3, in place of the 18-kb stretch of chromosome encoding the msm and GAL operons. Strains that were also unable to utilise beta-glucosides were found to have a separate 4 kb deletion in the BGL regulon that is proposed to be due to homologous recombination between two short stretches of identical sequence. The evidence is consistent with all the melibiose-negative strains examined being derived from a common ancestor. PMID- 12652054 TI - Influence of apolipoprotein e polymorphism on cardiovascular risk factors in obese children. AB - AIM: The main objective of the study was to determine whether risk factors associated with obesity are influenced by genetic variation of apolipoprotein E (ApoE). METHODS: 81 obese children (mean age 9.4 +/- 2.8 years) and an age matched control group were included. Body composition, lipid profile, and glucose and insulin levels were evaluated according to international recommendations, and the blood pressure was measured by an oscillometric method. RESULTS: The calculated frequencies of the ApoE alleles *2, *3, and *4 (0.04, 0.88 and 0.08) in obese children were similar to those of eutrophic age-matched controls (0.07, 0.82, and 0.11) and fitted the range of variation generally observed in southern European populations. Age, anthropometric parameters, body fat mass, and blood pressure were similar in E2/3, E3/3 and E4/3 genotypes. Total/high-density lipoprotein cholesterol and low-density lipoprotein/high-density lipoprotein cholesterol ratios were higher in the E4/3 group as compared with E3/3 (p < 0.01) and E2/3 (p < 0.05) groups. No differences concerning clusters of risk factors were observed among the three genotypes. No associations were found between ApoE polymorphism and glucose levels (fasting and at 2 h) and between fasting insulin levels and HOMA(IR) results. Higher levels of fasting and 2-hour insulin and higher HOMA(IR) values were significantly associated with a higher fat mass. CONCLUSIONS: ApoE polymorphism seems to influence some lipid profile abnormalities associated with obesity in childhood. However, clustering of risk factors and insulin resistance seem not to be dependent on ApoE polymorphism. PMID- 12652055 TI - Weekly high-dose folic Acid supplementation is effective in lowering serum homocysteine concentrations in women. AB - AIMS: To compare the efficacy of a weekly high-dose (2,800 microg) folic acid supplement with a daily (400 microg) folic acid supplement in lowering homocysteine concentrations in healthy women of childbearing age. METHODS: Free living healthy women of childbearing age (n = 138) were randomized to receive a weekly (2,800 microg), a daily (400 microg) folic acid supplement, or placebo. Blood samples were collected at baseline and at 12 weeks and analyzed for serum homocysteine and erythrocyte folate. RESULTS: At 12 weeks the mean (95% CI) serum homocysteine concentrations declined significantly (p < 0.05) in women receiving the daily [week 12 - baseline: -1.4 (-2.0, -0.70) micromol/l] and the weekly supplement [-1.3 (-2.1, -0.5) micromol/l] versus placebo [0.0 (-0.5, 0.5) micromol/l]. There was no significant difference between the two folate-treated groups (p > 0.05). At 12 weeks the mean erythrocyte folate concentration increased significantly in both supplemented groups versus placebo (p < 0.001). The increase in erythrocyte folate was significantly greater (p < 0.001) in the daily group than in the weekly group [451 (380, 521) vs. 288 (240, 335) nmol/l]. CONCLUSION: A weekly high-dose folic acid supplement was as effective as a daily supplement in lowering homocysteine concentrations in healthy women of childbearing age. Further study is needed to determine if weekly folic acid supplementation is effective in lowering homocysteine concentrations in populations with high homocysteine. PMID- 12652056 TI - Iron and folate status before pregnancy and anemia during pregnancy. AB - BACKGROUND: A lack of information about the role that iron and folate stores (erythrocyte folate) play on the anemia incidence in pregnant women. OBJECTIVE: To evaluate the predictive value of ferritin and erythrocyte folate before pregnancy in relationship to the incidence of anemia throughout pregnancy. METHODS: Thirty-five non-anemic non-pregnant women were evaluated 1 or 2 months before the pregnancy. After becoming pregnant they were evaluated every 4 weeks starting in the 8th week of pregnancy until 1 month post-partum. Every evaluation included determination of hemoglobin, ferritin and erythrocyte folate. RESULTS: When the women started gestation with an insufficient iron store (<20 microg/l), the trend was to present anemia at significantly earlier gestational ages, in such a way that at week 20 of gestation almost 60% of the women with an inadequate store had anemia and only 25% of the women with an adequate store had this condition. On the other hand, the pregestational erythrocyte folate did not predict anemia incidence. CONCLUSIONS: Pregestational ferritin concentration predicts value of incidence of anemia throughout pregnancy. PMID- 12652057 TI - Polyphenol pattern and antioxidant activity of different tomato lines and cultivars. AB - BACKGROUND/AIMS: Besides antioxidant vitamins and minerals, fruits and vegetables contain flavonoids and related phenolics. The biological activities of these polyphenols have become well known in recent years evidencing their beneficial effects on human health. In this context, the characterization of the flavonoids present in tomatoes is of great interest. Thus the polyphenol pattern (including flavonols, flavanones and cinnamate derivatives), lycopene and beta-carotene concentrations and the total antioxidant activity (TAA) of the phenolic fraction from different tomato lines and cultivars have been determined. METHODS: The characterization was obtained by means of spectrophotometry and HPLC analyses. RESULTS: Mean values for single flavonoids were 0.68 +/- 0.16 for naringenin, 0.74 +/- 0.12 for rutin and 0.32 +/- 0.06 for a rutin-pentoside. Mean total polyphenol content was 13.15 +/- 1.15 mg/100 g and mean TAA value was 1.3 +/- 0.10 mmol/g. The obtained TAA values resulted in good accordance with the total polyphenol content (R(2) = 0.7928). The main phenolic acids were chlorogenic (mean +/- SE 0.20 +/- 0.03) and caffeic acid (mean +/- SE 0.03 +/- 0.01). Mean levels of lycopene and beta-carotene were 5.38 +/- 0.90 and 1.18 +/- 0.40 mg/100 g, respectively. CONCLUSIONS: Almost all the lines characterised by low carotenoid content produce high levels of polyphenols, and consequently have the most powerful antioxidant potential. PMID- 12652058 TI - Isoflavone consumption does not increase the bone mass in osteopenic obese female zucker rats. AB - Some controversy exists in the literature concerning the effects of leptin on bone metabolism. Thus we have compared femoral bone density and biochemical markers of bone metabolism in male and female fatty (leptin-resistant) Zucker rats and their lean homozygous controls at 3 and 6 months of age. At 3 months, no differences concerning total, diaphyseal (cortical bone), and distal metaphyseal (trabecular bone) femoral bone densities, plasma osteocalcin concentrations, and urinary deoxypyridinoline excretion were observed between fatty and lean rats. On the opposite, at 6 months of age, in both males and females, total, diaphyseal, and distal metaphyseal femoral bone densities and plasma osteocalcin concentrations were lower in Zucker than in lean rats. Soybean isoflavone consumption (40 microg/g body weight/day for 90 days, a dose which prevents osteopenia following ovariectomy both in lean Zucker homozygous controls and in Wistar rats) by obese female Zucker rats had no significant effect upon their bone mass. PMID- 12652059 TI - Wheat bran and oat bran effectively reduce oxidative stress induced by high-fat diets in pigs. AB - AIM: To evaluate the influence of wheat bran and oat bran on the oxidative stress induced by a high proportion of fat in the diet. METHODS: Forty-eight growing pigs were penned individually and after an adaptation period divided into four groups. All groups received isocaloric daily rations composed of basal diet which was then supplemented with: starch (controls; CONT), linseed oil (OIL), linseed oil and wheat bran, or linseed oil and oat bran. The experimental period lasted 14-days. The oxidative stress was evaluated by measuring the malondialdehyde (MDA) concentration in blood plasma, the 48-hour urinary MDA excretion, and the degree of leukocyte nuclear DNA damage. RESULTS: In comparison with the CONT group, a significant increase in the MDA concentration in blood plasma and in the MDA excretion in urine was found in the OIL group. The degree of DNA damage in the OIL group was also significantly higher. In comparison with the OIL group, the wheat bran and oat bran supplementation significantly reduced the 24-hour MDA excretion in urine and reduced the degree of DNA damage in leukocytes to the level of the CONT group. CONCLUSION: The results of the experiments confirmed that a high wheat bran and oat bran intake effectively reduces oxidative stress induced by a high-fat diet. PMID- 12652060 TI - Nutritional and clinical evaluation of a modified soy protein with covalently bound branched-chain amino acids in cirrhotic sprague-dawley rats. AB - Several studies suggested that branched-chain amino acids (BCAA) improve plasma amino acid imbalance as well as protein metabolism in patients with cirrhosis. However, commercial formulas supplemented with free BCAA have their limitations. We evaluated a modified soy protein diet with covalently bound BCAA (diet M) by comparing it with diets based on casein (diet C) or Hepatic Aid II (diet H; commercial formula) as protein sources. After 3 weeks of bile duct obstruction, 24 Sprague-Dawley rats divided into three groups received diets with 9% (w/w) protein/amino acids for 7 days. Nutritional and clinical parameters were determined. Nitrogen balance and weight gain (g)/protein intake (g) with diet M (0.19 +/- 0.31 and 1.33 +/- 1.43 g, respectively) were significantly higher (p < 0.05) than with diet H (-0.34 +/- 0.20 and -0.34 +/- 1.11 g), but comparable to those with diet C (0.04 +/- 0.38 and 0.20 +/- 0.93 g). Animals on diet M had a significantly (p < 0.05) increased plasma BCAA:aromatic amino acid ratio (1.8 +/- 0.3) as compared with those on diets H (1.3 +/- 0.1) and C (0.8 +/- 0.0). There were no significant differences in organ weight or liver function among the groups. We conclude that the BCAA-modified protein is an attractive option in the nutritional support of patients having cirrhosis. PMID- 12652061 TI - Viscachataenia n. g., with the redescription of V. quadrata (von Linstow, 1904) n. comb. (Cestoda: Anoplocephalidae) in Lagidium viscacia (Rodentia: Chinchillidae) from Argentina. AB - A new genus, Viscachataenia, is erected to accommodate the anoplocephalid cestode Cittotaenia quadrata von Linstow, 1904, based on a redescription of the species from material collected from the chinchillid rodent Lagidium viscacia in Argentina. The new genus is characterised by paired genitalia, a reticulate uterus and the vagina entering the genital atrium anterior to the cirrus-sac. Viscachataenia therefore has similarities with Monoecocestus Beddard, 1914, a genus which is common in South American rodents but which has a single set of genitalia in each segment. Cittotaenia viscaciae (Spasskii, 1951) and Bertiella findlayi Mazza, Parodi & Fiora, 1932, also from viscachas, are considered synonyms of V. quadrata. PMID- 12652062 TI - Catatropis chilinae n. sp. (Digenea: Notocotylidae) from Chilina dombeiana (Gastropoda: Pulmonata) and notes on its life-cycle in Patagonia, Argentina. AB - A new species of Catatropis from a freshwater pulmonate snail of the family Chilinidae, which is endemic to South America, is described. Naturally infected Chilina dombeiana were collected from several localities in Andean Patagonia. The characteristics of the larval stages are presented. Experimentally reared adults, located in the distal portion of the intestinal caeca, were recovered from chickens and ducks. Adults of Catatropis chilinae n. sp. can be distinguished from all other species in having 9-11 (10) ventral glands, a cirrus-sac extending between the first third and the middle of the body, a metraterm slightly shorter or equal to the cirrus-sac, vitelline follicles reaching forward to the middle of the body, lobed testes, and a genital pore closely posterior to the caecal bifurcation. Eggs bear polar filaments only at the anopercular end. Rediae have only one or two cercariae. Shed cercariae are trioculate with a long tail and encyst in the environment, and metacercariae become infective 72 hours after encystment. This species is widely distributed between 40 degrees 10' S and 43 degrees 09' S and it is the first Catatropis species recorded for the Chilinidae and for Argentina. PMID- 12652063 TI - Scorpidotrema longistipes n. g., n. sp. (Digenea: Opecoelidae) from Scorpis georgiana (Teleostei: Scorpididae) from southern Western Australia. AB - Scorpidotrema longistipes n. g., n. sp. is described from the intestine of Scorpis georgiana Valenciennes (Scorpididae) from off Point Peron, Western Australia. The new genus is distinguished by the combination of a remarkably long and retractable ventral sucker peduncle, a possible uroproct, well-developed cirrus-sac and a uterine seminal receptacle. The subfamilial relationships of the new genus are troublesome. It incorporates features of the Opecoelinae, Stenakrinae and Plagioporinae. The absence of a canalicular seminal receptacle suggests a relationship with the Opecoelinae and Stenakrinae, whereas the well developed cirrus-sac suggests a relationship with the Plagioporinae and Stenakrinae. The overall arrangement of the gonads is not similar to that of existing genera of Stenakrinae. It is concluded that the genus is best placed in the Stenakrinae although that subfamily may now be an artificial assemblage. This new genus forms part of a distinctive fauna of trematodes restricted to Australian southern temperate fishes. PMID- 12652064 TI - Monogenea from the middle Yangtze Valley: Sinomazocraes changjiangensis n. g., n. sp. (Family Mazocraeidae Price, 1936) on a clupeiform fish from Hubei Province, China. AB - Sinomazocraes changjiangensis n. g., n. sp. (Monogenea: Mazocraeidae) is described from the gills of the shortjaw tapertail anchovy Coilia brachygnathus collected from Jiayu and Huanggang Counties (Hubei Province), which are located in the middle reaches of the Yangtze River. The structure of the clamps and copulatory organ are similar to those of Heteromazocraes Mamaev, 1981. However, although Sinomazocraes also has four pairs of clamps, it is distinguished from other genera in the family by the fact that the three anterior clamps on one side of the body are significantly larger than the others. Variations in the form of the clamps in different species of the Mazocraeidae are discussed. PMID- 12652065 TI - A list of monogeneans from Chinese marine fishes. AB - This paper contains a brief history of studies on monogeneans of Chinese marine fishes and a list of 337 monogenean species, together with their marine hosts and notes on the sea in which they were found. An additional 139 species have been added to the 198 species recorded in 'Monogeneans of Chinese Marine Fishes' published by Zhang, Yang and Liu in 2001. PMID- 12652067 TI - Bisbalia vossi n. g., n. sp. (Nematoda: Onchocercidae), a filarial worm from a geomyoid rodent, Heteromys anomalus, in Venezuela. AB - Bisbalia vossi n. g., n. sp. is described from Heteromys anomalus (Rodentia: Geomyoidea: Heteromyidae) in northern Venezuela (Aragua). The filariae were found in a membranous pocket in the pleural cavity, and almost all had ingested red blood cells of their host. The morphology of this onchocercine species is highly evolved (advanced reduction of head and caudal papillae; short undivided oesophagus). Its very short microfilariae (60 microm) and the shape of the tail of the female (two terminal median pairs of bosses) suggest that this species could be derived from Ackertia Vaz, 1934, a South American genus parasitic in caviomorph rodents which is related to the Dipetalonema-line, but Ackertia has several pairs of precloacal papillae, which are absent in the new genus. In North America, where the geomyoid rodents originated and diversified, the two previously described filarial species differ from this new material and show affinities with Old World bat parasites (Litomosa van Beneden, 1871). PMID- 12652066 TI - Lecithostaphylus retroflexus (Molin, 1859) (Zoogonidae) and Tergestia acanthocephala (Stossich, 1887) (Fellodistomidae) (Digenea) from the epipelagic teleost Belone belone (L.) in the western Mediterranean. AB - Numerous individuals of the poorly known species Lecithostaphylus retroflexus (Zoogonidae) and Tergestia acanthocephala (Fellodistomidae) have been recovered from the teleost fish Belone belone gracilis from off the Scandola Nature Reserve, Western Mediterranean. They are redescribed, incorporating previously undescribed features: for L. retroflexus, a post-oral ring, a bipartite seminal vesicle, the shape of the excretory vesicle, the subterminal excretory pore and the flask-shaped gland-cells associated with the distinctly pedunculate ventral sucker; and for T. acanthocephala, the intestinal bifurcation in the forebody, necessitating its return to the genus Tergestia from Theledera. Additionally, T. acanthocephala is compared with T. laticollis from various species of Trachurus from the same geographical area. PMID- 12652068 TI - Allopodocotyle skoliorchis n. sp. (Opecoelidae: Plagioporinae) from Parequula melbournensis (Castelnau) (Gerreidae), a temperate marine fish in Australian waters. AB - A new species of Allopodocotyle Pritchard, 1966 is described from the intestine and pyloric caeca of Parequula melbournensis (Gerreidae) caught from the waters off South and Western Australia. The new species is distinguished from other species by its larger eggs, broader form, pre-bifurcal genital pore and a number of other measurable features that are discussed. Of the species that share morphological similarities with Allopodocotyle skoliorchis n. sp., it is the only species known from a gerreid; all the other species are from serranids. PMID- 12652070 TI - Taxonomic revision of Tatria Kowalewski, 1904 (Cestoda: Amabiliidae): redescriptions of T. biremis Kowalewski, 1904 and T. minor Kowalewski, 1904, and the description of T. gulyaevi n. sp. from Palaearctic grebes. AB - Two species of Tatria Kowalewski, 1904 are redescribed from grebes in Bulgaria: T. biremis Kowalewski, 1904 (specimens from Podiceps nigricollis) and T. minor Kowalewski, 1904 (specimens from P. cristatus and P. nigricollis). T. mircia Gulyaev, 1990 is synonymised with T. minor. The previous records of T. biremis, T. minor and T. mircia are critically analysed in view of the present results. T. gulyaevi n. sp. is described from P. nigricollis from Bulgaria and the Czech Republic and from an unidentified grebe species from Turkey. Some of the previous records of T. minor and T. biremis are recognised as belonging to T. gulyaevi. One specimen illustrated by Kowalewski (1904) is designated as a lectotype of T. minor in order to stabilise the nomenclatural standing of this species. PMID- 12652069 TI - Phylogenetic relationships of Echinostoma Rudolphi, 1809 (Digenea: Echinostomatidae) and related genera re-assessed via DNA and morphological analyses. AB - In order to investigate the relationships within the Echinostomatidae two data sets of gene sequences were analysed. The first consisted of all previously published ND1 sequences (20) together with 17 new sequences. The latter represented six species from the cosmopolitan genera Echinostoma, Echinoparyphium, Hypoderaeum and Isthmiophora. The second data-set of ITS sequences again included all previously published sequences (12) and three new sequences from species of Echinostoma, Echinoparyphium and Isthmiophora. All new isolates, as well as voucher material from five previously sequenced isolates, were identified on the basis of morphological characters. The phylogenetic trees inferred from the ND1 data set helped to clarify the generic affiliation of all isolates and confirmed the morphological identifications. The only exception was Echinoparyphium aconiatum, whose current position in the genus Echinoparyphium was not supported by the sequence data. Although the ITS data provided insufficient resolution for an unequivocal solution to the relationships within the genus Echinostoma, it supported the identification of Echinoparyphium ellisi and the distinct species status of three isolates of Echinostoma revolutum as predicted from the ND1 data. PMID- 12652071 TI - A new genus of hard ticks in Cretaceous Burmese amber (Acari: Ixodida: Ixodidae). AB - A hard tick larva in Cretaceous Burmese amber is described as Cornupalpatum burmanicum n. g., n. sp. Diagnostic characters include a subcircular body with a marginal groove, 11 festoons, elongate four-segmented palpi with the fourth segment distinct and apical, the absence of an anal groove and eyes, and the presence of claws on palpal segment 3. The last character is unique for all members of the Ixodida, both fossil and extant. Aside from the palpal claws and marginal groove, features of the tick larva closely resemble those of members of the genus Aponomma Neumann 1899, considered one of the most primitive tick lineages today, whose hosts are primarily reptiles. PMID- 12652072 TI - Redescription of some Indian species of Thaparocleidus Jain, 1952 (Monogenea), with aspects of the developmental biology and mode of attachment of T. pusillus (Gusev, 1976). AB - Thaparocleidus wallagonius Jain, 1952, the type-species of the genus, is redescribed from the type-host and locality. T. gomtius (Jain, 1952), T. seenghali (Jain, 1961), T. indicus (Kulkarni, 1969), T. malabaricus (Gusev, 1976) and T. pusillus (Gusev, 1976) were also redescribed, as their original descriptions are incomplete. The following new synonymies for some Indian species are proposed: Urocleidus bhagwanpurensis Agrawal & Singh, 1982, as a junior synonym of T. wallagonius; U. gorakhpurensis Agrawal & Singh, 1982, Murraytrema gorakhpurensis Agrawal & Singh, 1985 and Ancylodiscoides kulkarnii Agrawal & Sharma, 1988 as junior synonyms of T. seenghali; and U. tandani Agrawal & Singh, 1982, A. meerutensis Singh, Agarwal, Kumar & Vibha, 1997 and Silurodiscoides elongatus Singh, Agarwal, Kumar & Vibha, 1997 as junior synonyms of T. indicus. Data on the developmental biology and mode of attachment of T. pusillus from Mystus bleekeri at Lucknow are included. PMID- 12652073 TI - Naricolax insolitus n. sp., a bomolochid copepod (Poecilostomatoida) parasitic in the nasal cavities of silver pomfret Pampus argenteus off Taiwan. AB - A new species of bomolochid copepod (Poecilostomatoida), Naricolax insolitus n. sp., parasitic in the nasal cavities of the silver pomfret Pampus argenteus (Euphrasen) from the Strait of Taiwan is described. The new species is distinguished from its only known congener, N. atypicus Ho, Do & Kasahara, 1983, by possessing the following four apomorphic features: (1) a complex outgrowth on the egg-sac attachment area; (2) a bent egg-sac; (3) a sharp hook (modified seta V) on the caudal ramus; and (4) an armature of II,I,4 (instead of II,I,5) on the terminal segment of leg 4 exopod. This is the first record of parasitic copepod from the nasal cavities of the silver pomfret. PMID- 12652074 TI - Description of Pomphorhynchus spindletruncatus n. sp. (Acanthocephala: Pomphorhynchidae) from freshwater fishes in northern Iraq, with the erection of a new pomphorhynchid genus, Pyriproboscis n. g., and keys to genera of the Pomphorhynchidae and the species of Pomphorhynchus Monticelli, 1905. AB - The status of species of Pomphorhynchus Monticelli, 1905 is examined and 23 species are recognised as valid. These include P. spindletruncatus n. sp., which is described herein from two species of freshwater fishes in northern Iraq, Aspius vorax Heckel and Barbus xanthopterus (Heckel) (Cyprinidae). Only one other species, P. yunnanensis Wang, 1981, has a spindle-shaped trunk similar to that of P. spindletruncatus, but is distinguished from it by a distinctly different proboscis armature. A key separating the new taxon from other species of Pomphorhynchus is included. P. heronensis Pichelin, 1997 is reassigned to a new pomphorhynchid genus, Pyriproboscis n. g., based on its unique proboscis shape and armature, short proboscis receptacle and tubular cement glands. A key to the genera of the Pomphorhynchidae is also included. PMID- 12652075 TI - Binding of milk oligosaccharides by several enterotoxigenic Escherichia coli strains isolated from calves. AB - Milk oligosaccharides have been proposed to play an important role in newborn defense, blocking bacterial adhesion to the intestinal mucosa and preventing infections. Some studies have been performed on human milk oligosaccharides. Here we checked whether bovine milk oligosaccharides would achieve the same protective action against the most common calf enteric pathogens. Seven enterotoxigenic Escherichia coli strains, isolated from diarrheic calves, were selected. All strains managed to agglutinate horse erythrocytes, and we therefore used the inhibition of hemagglutination in the presence of oligosaccharides as an indicator of the union between oligosaccharide and bacterial adhesins. Oligosaccharides from different stages of bovine lactation and standard oligosaccharides were assayed. Midlactation milk, in particular that corresponding to the transition period, proved to be the most efficient at inhibiting hemagglutination. The standard oligosaccharides used pointed to the preference of several strains (K99-, F41-, and F17-fimbriated) for alpha2,6 linked sialic acid. By contrast, B23 fimbriae exhibited higher affinity for alpha2,3-sialylated isomers and B64 seemed to require N-acetylglucosamine for binding. Our results suggest a general trend for milk oligosaccharides. Probably they participate in the protection of newborn mammals from pathogens. PMID- 12652077 TI - Structural studies on IgG oligosaccharides of patients with primary Sjogren's syndrome. AB - Sjogren's syndrome (SS) is an autoimmune disease, and some patients have been found to have SS complicated with rheumatoid arthritis (RA), in which IgG is known to carry abnormal N-linked oligosaccharides. In order to investigate the relationship between SS and RA, the structures of N-linked oligosaccharides of IgG from 12 primary SS patients without RA, 9 RA patients, and 8 healthy individuals were analyzed using reversed-phase high-performance liquid chromatography, in combination with sequential exoglycosidase treatment and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. All of the IgG samples obtained from primary SS patients, RA patients, and healthy individuals contained the same series of biantennary complex-type oligosaccharides, but the ratio of each oligosaccharide differed among these 3 groups. The incidence of galactose-lacking N-linked oligosaccharides obtained from the IgG of RA patients was significantly higher than that from healthy individuals, but that from the serum IgG of primary SS patients varied among individuals. The patients with primary SS were classified into two groups based on the galactosylation levels of IgG oligosaccharides; one group exhibits galactosylation levels as low as those of RA patients and another exhibits levels similar to those of healthy individuals. Measurement of levels of rheumatoid factor (RF) revealed that primary SS patients with a high incidence of RF belonged to the low galactosylation group, as did RA patients. These results suggest that appearance of IgG carrying abnormal N-linked oligosaccharides in primary SS may be related to future complication with RA. PMID- 12652076 TI - The expression of human FUT1 in HT-29/M3 colon cancer cells instructs the glycosylation of MUC1 and MUC5AC apomucins. AB - Recently, we have reported that in normal gastric epithelium, the expression of gastric apomucins MUC5AC and MUC6 is associated with the specific expression of type 1 and type 2 Lewis antigens, and FUT2 and FUT1 fucosyltransferases, respectively. Until now, there are no data demonstrating the direct implication of specific glycosyltransferases in the specific patterns of apomucin glycosylation. HT29/M3 colon cancer cell line express MUC1, MUC5AC, type 1 Lewis antigens and FUT2 but not type 2 structures and FUT1, as it occurs in the epithelial cells of the gastric superficial epithelium. These cells were transfected with the cDNA of human FUT1, the alpha-1,2-fucosyltransferase responsible for the synthesis of type 2 Lewis antigens, to assess the implication of FUT1 in the glycosylation of MUC1 and MUC5AC. The M3-FUT1 clones obtained express high levels of type 2 Lewis antigens: H type 2 and Ley antigens. Immunoprecipitation of MUC1 and MUC5AC apomucins gives the direct evidence that FUT1 catalyses the addition of alpha-1,2-fucose to these apomucins, supporting the hypothesis that the pattern of apomucin glycosylation is not only instructed by the mucin primary sequence but also by the set of glycosyltransferases expressed in each specific cell type. PMID- 12652078 TI - Glycosylation and over-expression of endometriosis-associated peritoneal haptoglobin. AB - Peritoneal endometriotic tissues synthesize and secrete haptoglobin (pHp), which has an analogous nucleotide sequence to hepatic haptoglobin found in serum (sHp). This study performed enzymatic digestions and lectin binding assays to determine if differences in protein glycosylation exist between sHp and pHp, which may provide insight into pHp function and/or identify epitopes for development of novel methods of medical management of endometriosis. To reduce the dependence on surgical collection of peritoneal tissues from women, recombinant peritoneal Hp (rpHp) was produced and its glycosylation analyzed for future functional studies. These results showed the apparent molecular weight of pHp was 3 kDa smaller than sHp. Desialylation and complete N-deglycosylation elicited similar shifts in sHp and pHp electrophoretic migration, suggesting similar sialic acid content and indicating the 3 kDa variance was due to carbohydrate content, not protein degradation, respectively. Sequential deglycosylation of the four sHp N-glycan chains caused a 3 kDa shift per N-glycan removed suggesting the 3 kDa difference between sHp and pHp may be one N-glycan chain. Lectin ELISA and lectin-blotting analyses demonstrated increased pHp and rpHp interactions with MAL and LTL but no difference in binding to SNL compared to sHp from healthy individuals, identifying variations in the ratios of alpha(2-3) to alpha(2-6) sialic acid and fucose residues. Recombinant pHp was 100-fold over-expressed with a similar glycosylation pattern to pHp, albeit in an unprocessed alpha-beta Hp polypeptide form. These results are the first to identify differences between pHp and sHp glycosylation and lay groundwork further studies to characterize anomalies in glycan composition and structure, which likely impart pHp with known immunomodulatory functions and may be used as epitopes for development of immune based therapeutics for novel, non-surgical management of endometriosis. PMID- 12652079 TI - An NMR study of O-glycosylation induced structural changes in the alpha-helix of calcitonin. AB - We previously reported that two out of seven artificially O-glycosylated calcitonin derivatives had an altered peptide backbone conformation as indicated by decreased helical contents, determined by CD measurement. In the present study, two of those derivatives, in which a GalNAc residue is attached to Thr6 or Thr21 of calcitonin, were analyzed by NMR in order to determine the structural changes induced by the O-glycosylation in more detail. Deviations in the chemical shifts suggest that the structural change is not global but only a local one and is located in the vicinity of each O-glycosylation site. The intensities of the NOE cross peaks, an indicator of alpha-helical structure, also were decreased around the O-glycosylation site. The hydrogen/deuterium exchange rates of the main chain amide protons increased at the N- or C-terminal portion of the alpha helix corresponding to the respective O-glycosylation site and explains the results of the CD experiments. The inter-residual NOE cross peaks between the carbohydrate and the peptide portions, other than the O-glycosylated amino acid residue, showed that local structural contacts extended three or two residue distance for Thr6- or Thr21-glycosylated derivative, respectively. Thus, we conclude that the O-glycosylation induced a change in the local structure and that this structural perturbation modulated the original alpha-helical structure of calcitonin, resulting in the apparent decrease in the helical content deduced from CD spectra. PMID- 12652080 TI - Structure of the main ganglioside from the brain of Xenopus laevis. AB - The main component of the ganglioside(1) mixture from the brain of the adult amphibian Xenopus laevis accounts for 35% of the total, as lipid bound sialic acid. This ganglioside has been purified and characterized by thin layer chromatography, partial and exhaustive enzymatic hydrolysis with sialidase, TLC overlay procedures with anti-Gg(4)Cer and anti-Neu5Acalpha6GalNAcbeta specific monoclonal antibodies and mass spectrometry. All together the results suggest the following structure: Neu5Acalpha8Neu5Acalpha3Galbeta3(Neu5Acalpha8Neu5Acalpha6)GalNAcbeta4Galbeta4Glcb ta1Cer, or, IV(3)-alpha-Neu5Ac(2),III(6)-alpha-Neu5Ac(2)-Gg(4)Cer. PMID- 12652082 TI - Two glycosylation alterations of mouse intestinal mucins due to infection caused by the parasite Nippostrongylus brasiliensis. AB - The glycosylation alterations of mouse small intestinal mucins during a 12-day infectious cycle caused by the parasite Nippostrongylus brasiliensis have been studied. The guanidinium chloride insoluble mucins were isolated at day 0 to 12 from the small intestine of infected and non-infected C57BL/6 mice. The O-linked oligosaccharides were released by reductive beta-elimination from the mucins and separated into neutral, sialylated and sulfated fractions. All fractions were analyzed by monosaccharide composition analysis and the neutral oligosaccharides were structurally characterized by gas chromatography/mass spectrometry. Two oligosaccharides containing blood group H-type epitopes (Fucalpha1-2Gal-) were transiently expressed with a maximum at day 6. Additional oligosaccharides with the common structure HexNAc-Gal-3GalNAcol were transiently induced with a maximum at day 10. Northern blot analysis on total RNA showed a transient expression at day 4-6 of the Fut2 gene encoding a Fucalpha1-2 fucosyltransferase, probably responsible for the detected blood group H-type epitopes. Comparisons with the corresponding infection in rat studied previously, revealed structurally different alterations, although occurring as transient events in both species. Both showed an induced blood group-type transferase halfway through the infection (a blood group A transferase in rat) and an induced transferase adding a terminal GalNAc (to a sialic acid- containing epitope in rat) towards the end of the infection. These differences between closely related species suggest rapid evolutionary alterations in glycosyltransferase expression. PMID- 12652081 TI - Characterization of T cell hybridomas raised against a glycopeptide containing the tumor-associated T antigen, (betaGal (1-3) alphaGalNAc-O/Ser). AB - T cell hybridomas were raised against the glycopeptide S(72) (Core-1) containing the tumor-associated disaccharide betaGal (1-3) alphaGalNAc (Core-1) O-linked to serine at position 72 in the mouse hemoglobin derived decapeptide Hb (67-76). All hybridomas recognized the glycopeptide S(72) (Core-1). Two of the selected hybridomas responded, however, much better to the S(72) (Tn) glycopeptide containing the monosaccharide alphaGalNAc O-linked to serine. In addition, one hybridoma cross-responded to the glycopeptide T(72) (Core-1) having a threonine at position 72 instead of a serine. No cross-responses were found to other glycopeptides consisting of the same hemoglobin peptide with different glycans attached or to the unglycosylated peptides. The T cell receptor Valpha and Vbeta usage was clearly diverse. The CDR3alpha regions demonstrated moreover a predominance of small polar amino acid side chains, and three hybridomas contained a common sequence motif. All the sequenced CDR3beta regions contained furthermore a conserved proline-glycine motif. In conclusion, immunization with the disaccharide containing glycopeptides S(72) (Core-1) created a heterogeneous population of glycopeptide specific T cells with the ability of cross-responding toward related glycopeptides. PMID- 12652084 TI - Structural basis for the spotted appearance of amphibian neuromuscular junctions stained for synaptic vesicles. AB - We have compared the distribution of vesicles in amphibian motor nerve terminals determined by electron microscopy and by functional labeling with the styryl dye, FM2-10. Our aim was to resolve apparent discrepancies in the literature on the distribution of vesicles determined by the two procedures. Electron photomicrographs of non-serial cross sections of terminal branches were analyzed by stereological procedures to obtain indices of the terminal and vesicle areas. Terminal cross sectional area varied 3-fold on average along terminal branches and was largest particularly when active zone was present in the section. The vesicle area index (a measure of vesicle abundance) was highly correlated with the terminal area index, suggesting that the average density of vesicles is constant throughout the branches. When the data were separated according to whether active zone was present or not in a section, we found a small (26%) but significant increase in the average density of vesicles in active zone compared with non-active zone regions in the terminal. The distribution of spots along terminal branches following vesicle staining with FM2-10, as well as with antibodies to vesicle proteins, suggested that vesicles were distributed in highly concentrated clusters. However, the degree of variation between spot and inter-spot staining intensities found with the FM-dye was similar in magnitude to that for terminal cross sectional area determined from the electron microscopy. We conclude that the spotty pattern of stained vesicles seen with the optical microscope results primarily from vesicle accumulations associated with terminal varicosities. PMID- 12652083 TI - Characterization of human nerve basal lamina for their binding properties of anti MAG antibodies. AB - The functional importance of the basal lamina in Schwann cell development and in adult peripheral nerve fibers is well known. We have demonstrated previously by confocal microscopy that IgM deposits are present on the basal lamina of myelinating Schwann cells of nerve biopsies from patients with an anti-MAG IgM neuropathy. Therefore, the basal lamina was postulated to represent an early target for the uptake of autoantibodies on the surface of myelinated nerve fibers. In this study, the preparation of cell- and myelin-free basal lamina from human peripheral nerves, using a detergent-dependent method is described and characterized by immunohistochemical and biochemical analysis. Using these methods we demonstrated that an enrichment of basal lamina components of Schwann cells with extraction of myelin could be achieved. Western blot analysis and immunohistochemical characterization showed that anti-MAG IgM antibodies did not recognize an epitope on the basal lamina of normal nerves. The established method will allow in situ investigations of basal lamina components from human peripheral nerves in health and in disease, e.g. peripheral neuropathies of infectious or inflammatory origin. PMID- 12652086 TI - Different Ca2+ dynamics between isolated hippocampal pyramidal cells and dentate granule cells. AB - The hippocampal formation contains a variety of neuronal types. The principal neurons are granule cells in the dentate gyrus and pyramidal cells in Ammon's horn. These two neuron types show distinct cell morphology and display a different vulnerability to ischemic injury or various neurotoxins. In order to illustrate the difference in the pathophysiological properties of these neurons, we established a method for separately culturing granule cells and pyramidal cells. They were prepared from the dentate gyrus and Ammon's horn of 3-day-old Wistar rat pups and maintained for 7-9 days in culture. After transient exposure to N-methyl-D-aspartate or glutamate, both the cultured neuron populations displayed somatic Ca(2+) transients with similar amplitudes, but the subsequent recovery to baseline was about twice as fast in granule cells than in pyramidal cells. Similar results were obtained for K(+) depolarization-induced Ca(2+) elevation, suggesting that the relatively rapid Ca(2+) clearance in granule cells is independent of Ca(2+) influx pathways. The present study provides the first evidence for a difference in Ca(2+) dynamics and homeostasis between granule and pyramidal cells and may represent a cellular basis for the differential vulnerability of hippocampal neurons. PMID- 12652085 TI - In situ characterisation of oligodendrocyte progenitor cells in adult mammalian optic nerve. AB - Oligodendrocyte progenitors from the adult rodent optic nerve were described in vitro more than a decade ago but their identification in vivo has been more elusive. We have previously traced newly-generated oligodendrocytes remyelinating focal demyelination of adult feline optic nerve back to their origin in surrounding fascicles. Extending the search to normal tissue has now enabled the description of an oligodendrocyte progenitor population in situ. By combining serial 1 microm section immunocytochemistry with electron microscopy of perfusion fixed tissue, every cell of interest could be characterised. Oligodendrocyte progenitors were readily identifiable in transverse and longitudinal sections dual-stained with antibodies to glutamine synthetase and either S-100 or HNK-1. Oligodendrocyte progenitors of the adult feline optic nerve were mainly located centrally within fascicles, comprised 4% of the total macroglia and had both ultrastructural and immunocytochemical features suggesting roles additional to that of a progenitor cell. The identification and characterisation of oligodendrocyte progenitors in situ in fixed tissue sections should help the understanding of their role in the adult CNS and in disease such as multiple sclerosis. PMID- 12652087 TI - Application of potassium chloride to the surface of the rat visual cortex differentially affects the expression of presumptive neuroprotective molecules. AB - The purpose of this study was to investigate the effects of topical application of 3 M KCl for 20 minutes to the surface of the rat visual cortex upon the expression of presumptive neuroprotective molecules. Conventional immunohistochemical and immunoblotting techniques revealed a marked reduction of the expression of calbindin and parvalbumin after application of KCl, whereas calretinin, nitric oxide synthase, and the alpha7 subunit of the nicotinic acetylcholine receptor clearly increased following that procedure. Such effects were quantified both in terms of the number of immunoreactive perikarya and optical density of immunoblottings, and were observed as soon as 1 h, and to last at least until 15 days, following KCl application. These results suggest that calbindin and parvalbumin may not be able to exert an appreciable neuroprotective effect in the presence of KCl. On the other hand, calretinin, nitric oxide synthase, and the alpha7-containing nicotinic receptors are possibly upregulated to protect the neurons in which they are expressed. PMID- 12652088 TI - Microglial changes accompanying the promotion of retinal ganglion cell axonal regeneration into peripheral nerve grafts. AB - Intravitreal injection of the microglia inhibitor tuftsin 1-3 leads to an increase in retinal ganglion cell axonal regeneration into peripheral nerve grafts and a decrease in phagocytic cells in the retina. However, the relation of phagocytic cells and particularly microglia towards axonal regeneration remains unclear. Initially, to assess this, tuftsin 1-3's effect on axonal regeneration was reexamined by doing a dose-response study. Optimal doses were found to be 2.5 microg/ml and 250 microg/ml in rats and hamsters respectively. We then studied retinal phagocytic cells in rats. Microglial cells were classified as resting or activated based on their morphology following OX42 immunolabelling. In controls, most microglial cells were in the resting state. Optic nerve cut led to an increase in the total number of microglia and a ten-fold elevation in the proportion of activated cells; changes were more pronounced at the optic nerve stump. Anastomosis of an autologous segment of sciatic nerve to the stump of the freshly cut optic nerve minimized the overall increase in microglia, and combined with 2.5 microg/ml tuftsin 1-3, lead to a marked blunting of activation. Preservation within the retina of a higher proportion of resting over active form of microglia, and not the prevention of microglial proliferation per se, may be a crucial factor in allowing additional retinal ganglion cell axons to regenerate into peripheral nerve grafts. PMID- 12652089 TI - Chromatolysis of A-cells of dorsal root ganglia is a primary structural event in acute acrylamide intoxication. AB - To test the hypothesis that a somatofugal wave of atrophy moving distally in the axon of primary sensory neurons leads to loss of myelinated nerve fibers in acrylamide neuropathy, rats (N = 18) were intoxicated with an initial dose of 75 mg acrylamide per kg body weight followed by daily treatment with 30 mg/kg for three, six and 12 days. Ten age matched saline treated rats served as controls. Numbers and mean volumes of A- and B-cell perikarya of the L5 dorsal root ganglion, numbers of myelinated axons and the mean cross sectional myelinated axon area 3 and 18 mm from the ganglion in the dorsal root and in the sural nerve were estimated using stereological techniques. After three days no changes in the number or size of primary sensory perikarya or myelinated axons were observed. However, after six days 11% of the A-cell perikarya showed signs of chromatolysis (P < 0.001). After 12 days the rats showed signs of ataxia and 23% (P < 0.001) of A-cell perikarya were chromatolytic. There was a tendency for atrophy of the mean perikaryal volume of A-cells (2P = 0.059). The size-frequency distributions of axonal area of myelinated fibers in the dorsal root 3 mm from the ganglion were displaced to the left towards smaller sizes (25-50% quartile: 2P < 0.005 and 75 100% quartile: 2P < 0.05). In conclusion, the primary structural event in acute acrylamide intoxication is chromatolysis of A-cells of the dorsal root ganglion without the occurrence of somatofugal axonal atrophy. PMID- 12652090 TI - Long-term acrylamide intoxication induces atrophy of dorsal root ganglion A-cells and of myelinated sensory axons. AB - We have examined the effects of acrylamide on primary sensory nerve cell bodies and their myelinated axons in chronic acrylamide intoxication. The numbers and sizes of dorsal root ganglion cell bodies (L5) and myelinated nerve fibers were estimated with sterelogical techniques in severely disabled rats which had been treated with 33.3 mg/kg acrylamide twice a week for 7.5 weeks. There was no loss of dorsal root ganglion cells or myelinated nerve fibers in the roots, the sciatic nerve, sural nerve, and a tibial nerve branch. The mean perikaryal volume of A-cells was reduced by 20% (2P < 0.001) from 50000 microm(3) in controls (CV = 0.13) to 40000 microm(3) (0.12), whereas B-cell volume was unchanged. All size frequency distribution curves of myelinated axon area of peripheral nerves and sensory roots were shifted to the left towards smaller values in rats exposed to acrylamide. In the L5 sensory root 3 mm from the ganglion, there was a significant reduction of mean cross sectional area of myelinated axons by 14% (2P < 0.05) from 7.6 microm(2) (0.11) in controls to 6.5 microm(2) (0.13) in intoxicated rats. The mean cross sectional area of myelinated sural nerve axons was reduced by 22% (2P < 0.001) from 8.6 microm(2) (0.08) in controls to 6.7 microm(2) (0.17) in intoxicated rats. We conclude that chronic intoxication with acrylamide leads to selective atrophy of type A dorsal root ganglion cell bodies and simultaneous atrophy along their peripheral axons, whereas neuronal B-cell bodies and motor axons are spared. It is suggested that the neuronal atrophy might well represent a defect of neurofilament synthesis and transport. PMID- 12652091 TI - Randomized controlled trial on the effects of a skills-based workshop on medical students' management of problem drinking and alcohol dependence. AB - The purpose of this study was to determine whether a skills-based workshop will improve medical students' management of problem drinking and alcohol dependence in simulated patients. Seventy-six 3rd and 4th year Ontario medical students were randomized to receive a 3-h workshop on either problem drinking and alcohol dependence or depression (control condition). Students then completed eight simulated office visits (OSCE stations) with simulated patients presenting with depression, problem drinking or alcohol dependence. Examiners completed a checklist of the questions asked and advice given by the student, and simulated patients and examiners completed a global rating scale. Four months later, students were sent a survey on their knowledge, attitudes, and behavior towards patients with alcohol problems. The alcohol group received significantly higher assessment and management checklist scores and global rating scores than did the depression group (p < 0.01) and performed better on almost all aspects of clinical management of both problem drinking and alcohol dependence. On the follow-up survey (n = 55) the alcohol group showed a significant increase in beliefs about self-efficacy in managing alcohol problems (p < 0.05) and had greater knowledge of reduced drinking strategies, but the two groups did not differ on other measures. A skills-based workshop causes marked short-term improvements in medical students' management of problem drinking and alcohol dependence, an increase from baseline to postworkshop in self-efficacy beliefs that was sustained through to follow-up, and greater knowledge of reduced drinking strategies. Repeated reinforcement of clinical skills may be required for a long-term impact on clinical behavior. PMID- 12652092 TI - Gender differences in treatment-seeking Brazilian drug-dependent individuals. AB - The authors compared 66 women and 66 men entering an outpatient treatment for drug dependence, focussing on demographics, progression of dependence, and other associated clinical features. Most of the subjects were dependent on cocaine (75%). Women, as compared with men, had a higher level of education, were more likely to have migrated from other places to the city of Sao Paulo, to report past suicide attempts, and to have first-degree relatives with alcohol problems. There were no gender differences regarding the total time of drug use, baseline severity of dependence, and rates of concurrent psychiatric comorbidity. Clinicians working with drug-dependent women should note the high rates of suicidal behavior in this population, and should explore, besides psychiatric status, risk factors such as impulsive temperament traits. It should also be noted that higher rates of alcohol problems in women's family background pose challenges for the development of effective preventive and treatment strategies. PMID- 12652093 TI - The role of Medicaid HMO enrollment in the longitudinal utilization of medical care services in a cohort of injecting drug users in Baltimore, Maryland. AB - Although injecting drug users (IDUs) are highly dependent on Medicaid, the literature has not explored the effect of Medicaid HMO enrollment on medical care utilization. This longitudinal analysis compares 6-month utilization levels of IDUs in Medicaid HMOs with those of other Medicaid-enrolled IDUs and IDUs in other insurance arrangements. The analysis includes 1064 participants from the AIDS Link to Intravenous Experience (ALIVE) study, a natural history study of HIV infection in a community-based sample of IDUs in Baltimore, Maryland. A generalized linear modeling approach was used to examine self-reported inpatient admissions, ER visits, and ambulatory care visits from January 1995 through June 1997. Non-HMO Medicaid beneficiaries had more inpatient admissions and ambulatory care visits than Medicaid HMO enrollees. These findings suggest that HMO enrollment has been effective in lowering utilization among IDUs in Medicaid, but it is unclear whether utilization differences are the result of inappropriate utilization or access issues. PMID- 12652094 TI - Coordination of medical care and opioid dependence treatment in primary care: a case report. PMID- 12652095 TI - HDL genetics: candidate genes, genome wide scans and gene-environment interactions. AB - Cardiovascular disease (CVD) is multifactorial and its manifestation is determined by interactions among genes, as well as among genetic factors and numerous environmental factors. It has long been known that low levels of high density lipoprotein cholesterol (HDL-C) are associated with increased CVD risk. Moreover, the emerging data from developing countries, suggest that this may be the most common lipid abnormality observed in those societies. It has been clearly demonstrated that variation at several candidate genes has a significant effect over the spectrum of HDL-C levels observed in the general population. In addition, these effects are modulated by several non-modifiable such as gender and age, and modifiable factors, such as diet, smoking, obesity, and alcohol intake among many others. The lessons that we are learning from studying candidate gene-environment interactions should help us to chart the intricacies of the biochemical pathways involved in lipoprotein metabolism. Moreover, similar models need to be applied to the analysis of genome wide searches aimed to uncover new genes involved in HDL metabolism and reverse cholesterol transport. This knowledge should facilitate more targeted and effective public health policies, especially in developing countries, where the fast environmental changes are modifying the disease patterns by exposing a large number of susceptible individuals to the factors that have been responsible for the high prevalence of CVD in Western industrialized countries. PMID- 12652096 TI - Lipoprotein lipase deficiency--rare or common? AB - Cardiovascular disease is the number one killer in the world. Prevention is the most promising treatment. However, predicting an individual's risk for cardiovascular disease is quite complicated, involving the complex interaction of a plethora of environmental and genetic factors. Many of the environmental factors can be suppressed by adopting a healthier lifestyle; dietary modifications, increasing exercise, not smoking, etc. Understanding the role of many genetic factors remains somewhat elusive. Genes involved in lipid metabolism are a focus of this ongoing genetic research. There is substantial evidence to suggest that one such gene, lipoprotein lipase (LPL), has a profound influence on triglyceride and high-density lipoprotein (HDL) cholesterol levels in the blood; both independent risk factors for cardiovascular disease. Although a direct association has not be found, mutations resulting in LPL deficiencies have been linked to unfavorable lipid profiles. Although homozygous or compound heterozygous mutations in LPL resulting in complete loss of catalytically activity are rare, heterozygous mutations are quite common worldwide. It is when these moderate mutations are compounded by other risk factors, that a significant increase in risk of cardiovascular disease is observed. Therefore, it may be that these common deficiencies in LPL are an underlying cause for many incidents of cardiovascular disease. PMID- 12652097 TI - Familial hypercholesterolemia from children to adults. AB - An update on several aspects on familial hypercholesterolemia (FH) such as molecular basis, DNA diagnosis, clinical criteria for the diagnosis, treatment of adults and children and psychosocial aspects of FH is presented. PMID- 12652098 TI - Blockade of the Na+-Ca2+ exchanger is more efficient than blockade of the Na+-H+ exchanger for protection of the myocardium from lethal reperfusion injury. AB - Since the Na(+)-H(+) exchanger (NHE) is not the only pathway of Na(+) influx into cardiomyocytes during ischemia/reperfusion, we hypothesized that blockade of Na(+)-Ca(2+) exchanger (NCX) may be a more efficient strategy than is NHE inhibition for protecting the myocardium from infarction. To test this hypothesis, we compared KB-R7943 (KBR), a novel selective NCX blocker, with cariporide, a selective NHE blocker, with regard to their protective effects against infarction. In isolated rabbit hearts, infarction was induced by 30-min global ischemia/2-h reperfusion, and infarct size was determined by tetrazolium staining and expressed as a percentage of area at risk (%IS/AR). Hearts received no drugs, or were infused with cariporide (1 microM) for 10 min or KBR (0.3 or 10 microM) for 5 min before ischemia or after the onset of reperfusion. Protein level of NCX was assessed by Western blotting. Cariporide infusion before ischemia significantly reduced %IS/AR from 63.9 +/- 2.9% to 20.2 +/- 3.0%, but its infusion upon reperfusion failed to achieve a significant protection (%IS/AR = 53.8 +/- 4.6%). In contrast, KBR infusion similarly reduced infarct size both when infused before ischemia (%IS/AR = 33.3 +/- 6.3% and 21.9 +/- 4.7% by 0.3 and 10 microM KBR, respectively) and when infused for only 5 min after reperfusion (%IS/AR = 35.3 +/- 7.1% and 31.5 +/- 2.1% by 0.3 and 10 microM KBR, respectively). Protein levels of NCX after 30-min ischemia and 30-min ischemia/30 min reperfusion were similar to baseline values in both untreated controls and hearts treated with 0.3 microM KBR upon reperfusion. These results suggest that lethal reperfusion injury is more efficiently suppressed by blockade of the NCX than by blockade of the NHE. PMID- 12652099 TI - Dalteparin sodium (fragmin) administration following acute infarction does not affect myocardial perfusion and function in swine. AB - BACKGROUND: Heparin and low molecular weight heparin (LMWH) derivatives are considered angiogenic factors in ischemic and infarcted myocardium. However, the effect of dalteparin sodium (fragmin, a LMWH derivative) on angiogenesis post infarction has not yet been investigated.Purpose. To assess the effect of systemically-administered dalteparin sodium on left ventricular perfusion and function during remodeling in swine subjected to acute microembolization infarction (MI).Methods. MI was induced in 12 anesthetized female pigs which randomly received dalteparin sodium (12,500U) or placebo through intra-peritoneal osmotic pump, for one week, beginning immediately after the MI. Myocardial perfusion and function were measured using radionuclear scan and echocardiography, respectively, at baseline, immediately post MI, and at 2 and 4 weeks post MI. Dobutamine stress echocardiography was performed 4 weeks post MI. Serum level of basic fibroblast growth factor (bFGF) was analyzed using a commercially available ELISA kit.Results. No differences were observed in myocardial perfusion at all time points following MI. No significant changes were observed in myocardial function and wall motion during the 4 weeks follow-up period, or after dobutamine administration. Serum bFGF did not change throughout the study period.Conclusion. Under the setting of the current experiment, one week of dalteparin sodium administration does not affect myocardial perfusion and function following acute infarction in swine, either at rest or following pharmacological stress. PMID- 12652100 TI - Functional antagonism of different angiotensin II type I receptor blockers in human arteries. AB - OBJECTIVES: To evaluate and compare the functional type and the degree of antagonism of the selective angiotensin II type 1 receptor blockers (ARB) losartan, EXP 3174 (the active metabolite of losartan), valsartan and candesartan in human internal mammary arteries. METHODS: Human internal mammary arteries were obtained as excess graft material during coronary bypass surgery. Vessels were prepared as rings and mounted in an organ bath in which vasoconstriction and dilation can be measured. Concentration-response curves of angiotensin II mediated vasoconstriction were measured in absence or presence of different concentrations of one of the ARBs. RESULTS: Losartan showed a rightward shift of the angiotensin II-mediated vasoconstriction, whereas addition of its metabolite EXP 3174 caused a decrease of the maximal effect of angiotensin II. Incubation with valsartan and candesartan also resulted in a decrease of the maximal effect. The inhibiting effects on the angiotensin II-mediated vasoconstriction by the highest concentration of EXP 3174, valsartan and candesartan did not differ significantly. CONCLUSION: In human internal mammary arteries, losartan acts as a surmountable antagonist. On the other hand, EXP 3174, valsartan and candesartan demonstrate an insurmountable type of antagonism. Furthermore, the inhibiting effects of EXP 3174, valsartan and candesartan in our study are equal in the highest concentrations. PMID- 12652102 TI - The influence of magnesium on the electrophysiological effects of erythromycin in the isolated Guinea pig heart. AB - There have been several case reports that erythromycin can prolong the QT interval, resulting in torsades de pointes. As magnesium is well established in the treatment of torsades de pointes, we aimed to investigate how this trace element influences the electrophysiological effects of erythromycin. The effects of erythromycin and magnesium on cardiac conduction and refractoriness were evaluated in isolated guinea pig hearts perfused by the method of Langendorff. Erythromycin was given in concentrations of 10, 30 and 100 microM. In the magnesium group MgSO4 was elevated to 3.4 mM. Magnesium at a concentration of 3.4 mM did not affect cardiac conduction and refractoriness. Erythromycin prolongs the QT interval in a dose-dependent manner. The prolongation of the QT interval by erythromycin was diminished by the elevated magnesium concentration (3.4 mM) at high rates (pacing cycle length 180 vs. 220 ms). The most pronounced effect of erythromycin (100 microM) was the prolongation of the atrial effective refractory period. This effect was nearly abolished by the elevated magnesium concentration (129.0 +/- 8.3% vs. 41.9 +/- 10.6%, P < 0.01, n = 6, x +/- SEM). In summary, magnesium minimizes the effects of erythromycin on the repolarization period in the ventricle and on the effective refractory period of the atrium. PMID- 12652101 TI - Effects of benzyltetrahydropalmatine on two components of the delayed rectifier K+ current in Guinea pig ventricular myocytes. AB - The effects of benzyltetrahydropalmatine (BTHP), a new class III antiarrhythmic agent, on the action potential in guinea pig papillary muscle and the rapidly activating component (I(Kr)) and the slowly activating component (I(Ks)) of the delayed rectifier potassium current (I(K)) in isolated guinea pig ventricular myocytes were investigated. The action potentials of papillary muscles were studied using a standard microelectrode technique, while the K(+) currents were recorded using the whole-cell patch clamp technique. The results showed that BTHP prolonged the action potential duration (APD) without altering other variables of the action potential in guinea pig papillary muscles. The 2 components of I(K) were blocked by BTHP (1 approximately 100 micromol x L(-1)) in time-, concentration-, voltage-, and specifically frequency-dependent fashion. The IC(50) value for blockade of I(Kr) was 13.5 micromol x L(-1), while the IC(50) value for blockade of I(Ks) was 9.3 micromol x L(-1). BTHP 30.0 micromol x L(-1) reduced I(Kr) and I(Kr,tail) by 31 +/- 4.3% and 36 +/- 4.7% (n = 6, p < 0.01) and decreased I(Ks) and I(Ks,tail) by 40 +/- 6.3% and 39 +/- 4.6% (n = 7, p < 0.01) respectively. BTHP accelerated their deactivation course by reducing the time constants of deactivation of I(Kr) and I(Ks). The activation kinetics of I(Kr) or I(Ks) were not affected by BTHP. It is concluded that BTHP prolonged the action potential duration with respect to its non-selective action on I(Kr) and I(Ks) in single guinea pig ventricular cell in a frequency-dependent fashion. PMID- 12652103 TI - Effects of procainamide on transmural ventricular repolarisation. AB - To investigate the pharmacological effect of procainamide on transmural ventricular repolarisation in normal heart, the transmural activation-recovery intervals (ARI) and their responses to procainamide (20 mg/min i.v. for 20 min) were studied in 6 open-chest, pentobarbitone-anaesthetised sheep. ARI was measured from the unipolar ECGs acquired with 4 plunge needles inserted into the basal and apical parts of the left ventricular wall. During sinus rhythm (cycle length 500-700 ms), there was no significant difference in the pooled ARI between the epicardium (266.0 +/- 30.5 ms), midmyocardium (265.0 +/- 28.9 ms) and endocardium (265.7 +/- 28.1 ms) (p > 0.05). Procainamide prolonged ARI in all myocardial layers. The pooled ARI prolongation from the epicardium, midmyocardium and endocardium of the 6 animals was 66.8 +/- 18.3, 70.3 +/- 14.7 and 65.3 +/- 15.7 ms (p > 0.05), respectively. In conclusion, sodium channel blocker procainamide results in a similar repolarisation prolongation in the left ventricular epicardium, midmyocardium and endocardium of a healthy heart. PMID- 12652104 TI - Intima media thickness as a surrogate marker for generalised atherosclerosis. AB - Vascular markers of atherosclerotic burden allow atherosclerosis to be studied in the sub-clinical phase of the disease, and facilitate the assessment of new therapies for modifying coronary heart disease (CHD) risk factors. B-mode ultrasound enables non-invasive, direct visualization of the arterial wall. The intima media thickness of the carotid artery (CIMT) quantified using this technique, is a reliable marker of atherosclerotic burden. Furthermore, it demonstrates greater sensitivity in detecting early atherosclerosis compared with angiography. The validity of ultrasonographically-determined CIMT as a surrogate marker of atherosclerotic disease has been established; CIMT is positively associated with the incidence of coronary events, relates to atherosclerotic disease elsewhere in the arterial system and has common risk factors to the development of CHD. Furthermore, ultrasonographically-determined CIMT demonstrates good inter- and intra-observer reproducibility making it suitable for tracking the progression or regression of atherosclerotic disease over time. The validity of CIMT as a vascular marker of atherosclerosis progression has been demonstrated in clinical trials. Intervention studies have shown that modification of CHD risk factors can significantly reduce progression of CIMT. Hence, B-mode ultrasonography is being used increasingly to elucidate the efficacy of new therapies, enabling the benefits of treatment to be established more rapidly and with fewer patients compared with clinical outcome trials. PMID- 12652105 TI - Antihypertensive medication and quality of life--silent treatment of a silent killer? AB - Hypertension is one of the major killers in the modern world and one of the great ironies of the disease is that it has little or no immediate effect on affected individuals' quality of life (QOL). This makes treatment very difficult, as most medications will affect QOL negatively, either through side effects or through the change in life-style associated with chronic drug regimens. This review describes some of the conceptual background for QOL measurements and looks at the effects on QOL of different antihypertensive treatments available today. The latest addition to the range of antihypertensive drugs, angiotensin II-receptor blockers, may be a significant step forward in treatment, as their ease of dosing and excellent tolerability minimise negative effects on patients while their possible benefits on factors such as sexual function may even improve QOL in hypertensive patients. PMID- 12652106 TI - High dose lisinopril in heart failure: economic considerations. AB - BACKGROUND AND OBJECTIVES. In Switzerland, heart failure (HF) is one of the ten most common causes of hospitalization in patients older than 65 and the second most common in those over 80 years. 17% of the population in Switzerland is in this age class. The expenditures associated with HF are thus considerable. High doses of the angiotensin-converting enzyme (ACE) inhibitor lisinopril in patients with heart failure have been shown to significantly reduce the risk of death or hospitalization for heart failure and for any reason. The objective of the present study was to analyze the economic implications of using high doses of lisinopril in the Swiss health care system setting, both from the societal perspective and from that of the Swiss health insurers. PATIENTS AND METHODS: Using published data from the ATLAS Trial, we computed the monthly probability of death and of hospital admission for heart failure, ischemic events, other cardiovascular causes and any reason per patient for both arms of the trial. Swiss resource use patterns and Swiss unit costs were used to calculate expected monthly expenditures per patient from the third party payer perspective and from the societal viewpoint. Extensive sensitivity analyses were performed to test the robustness of our results. RESULTS: Expected monthly expenditures from the third party payer perspective are 347 Euro per patient in the low dose group and 319 Euro in the high dose group. This represents expected savings of 28 Euro per patient monthly. Because the public sector pays more than 50% of hospital costs, the savings from the societal viewpoint are greater. Expected monthly costs from the societal perspective are 762 Euro in the low dose group and 670 Euro in the high dose group per patient. This results in expected monthly savings of 92 Euro. Sensitivity analysis shows that the use of high doses of ACE inhibitor lisinopril does not result in higher health care expenditures for the whole system or even savings through a wide variety of practice patterns and unit costs. CONCLUSIONS: Based on the results of the ATLAS study, high doses of lisinopril are associated with a reduction in the number of hospitalizations for HF and for any reason and with a statistically non-significant reduction in mortality. The additional expenditures incurred because of prescription of high doses are offset by the savings in hospital costs. These are important considerations, given the increasing number of people aged 65 and older in Switzerland, and the prevalence of heart failure in this age group. PMID- 12652107 TI - Vitamins, homocysteine and cardiovascular risk. AB - A raised plasma level of the amino acid homocysteine is associated with increased risk of cardiovascular disease. This association may be causal-it is biologically plausible, fairly strong, graded, and an increase in plasma homocysteine preceeds the onset of vascular disease. Plasma homocysteine levels are controlled by genetic and nutritional factors, notably folate, vitamin B12 and vitamin B6 intakes. Folic acid in particular lowers plasma homocysteine levels by about 25%. It is not known if this cheap and safe treatment reduces vascular disease risk. Current randomized control trials are addressing this issue, and proof or otherwise of causality must await their results. Homocysteine may also interact with conventional risk factors such as smoking to substantially increase their effect on risk. Thus meticulous risk factor control may be particularly important in subjects at high total cardiovascular risk who also have a raised plasma homocysteine level, and folic acid supplementation may be considered in such individuals. PMID- 12652108 TI - Hyperhomocysteinemia, vascular function and atherosclerosis: effects of vitamins. AB - Homocysteine is a metabolic product of methyl group donation by the amino acid methionine. Moderate elevation of plasma homocysteine (>15 microM) is most commonly caused by B-vitamin deficiencies, especially folic acid, B(6) and B(12). Genetic factors, certain drugs and renal impairment may also contribute. Homocysteine has several potentially deleterious vascular actions. These include increased oxidant stress, impaired endothelial function, stimulation of mitogenesis, and induction of thrombosis. Homocysteine also appears to increase arterial pressure. In humans, experimental induction of hyperhomocysteinemia by methionine loading rapidly causes profound impairment of endothelium-dependent dilatation in both resistance and conduit arteries. This endothelial dysfunction can be reversed by administration of antioxidants. Epidemiological evidence suggests that homocysteine acts as an independent risk factor for atherosclerosis, thrombosis and hypertension. Prospective studies have shown that elevated plasma homocysteine concentrations in the top quintile of the population (>12 microM) increase risk of cardiovascular disease by about 2-fold. There are currently no data available from randomized, controlled trials of the effects of lowering plasma homocysteine on atherothrombotic events. Nonetheless, it would seem appropriate to screen for and treat hyperhomocysteinemia in individuals with progressive or unexplained atherosclerosis. Folic acid and vitamins B(6) and B(12) are the mainstay of therapy. Treatment of moderately elevated plasma homocysteine in patients without atherosclerosis should be deferred until the completion of randomized outcome trials. PMID- 12652109 TI - Vitamins, antioxidants and endothelial function in coronary artery disease. PMID- 12652110 TI - Antioxidant vitamins and risk of cardiovascular disease. Review of large-scale randomised trials. AB - People who consume a diet rich in fruit and vegetables have lower risks of cancer, cardiovascular disease and all-cause mortality. Many prospective cohort studies have reported inverse associations between dietary intake or blood levels of beta-carotene and risks of cancer. Several large-scale trials were set up to assess whether beta-carotene supplementation might reduce the risk of cancer. Subsequently, evidence emerged from basic research which indicated that oxidative modification of low-density lipoprotein cholesterol increases its atherogenicity. The evidence from basic research, and epidemiological evidence for a possible protective effect of antioxidant vitamins for cardiovascular disease was strongest for vitamin E. More recently, further trials were set up to examine if supplementation with anti-oxidant vitamins might also reduce the risk of cardiovascular disease. This review summarises the available randomised evidence from published trials of beta-carotene supplementation involving 70,000 people from 3 large-scale trials in healthy populations and on vitamin E supplementation involving 29,000 patients at high-risk of cardiovascular disease from 5 large scale trials. The results of these trials have been disappointing and failed to confirm any protective effect of these vitamins for either cancer or for cardiovascular disease. PMID- 12652111 TI - Comparison of the effects of cilostazol and milrinone on cAMP-PDE activity, intracellular cAMP and calcium in the heart. AB - We investigated the basis for the difference in the cardiotonic effects of the PDE3 inhibitors cilostazol and milrinone in the rabbit heart. Cilostazol displayed greater selectivity than milrinone for inhibition of cAMP-PDE activity in microsomal vs cytosolic fractions from rabbit heart. This difference was due to the inhibition of significantly less cytosolic cAMP-PDE activity by cilostazol compared to milrinone. A combination of cilostazol (>15 microM) and the PDE4 selective inhibitor, rolipram (5 microM), inhibited levels of cytosolic cAMP-PDE activity similar to those inhibited by milrinone on its own. This suggested that milrinone inhibited PDE4 in addition to PDE3 activity. In isolated rabbit cardiomyocytes, milrinone (>10 microM) caused greater elevations in intracellular cAMP and calcium than cilostazol. In the presence of rolipram, however, the cAMP and calcium elevating effects of cilostazol and milrinone were similar. Therefore, in rabbit heart, partial inhibition of PDE4 by milrinone contributed to greater increases in cardiomyocyte cAMP and calcium levels than cilostazol. PDE4 activity in failing human heart was lower than in rabbit heart and there was no significant difference in the inhibition of human cytosolic cAMP-PDE by cilostazol and milrinone. Our results suggest that in normal rabbit heart inhibition of PDE4 by milrinone may partly contribute to the greater cardiotonic effect of milrinone when compared to cilostazol. However, the lower level of PDE4 activity in failing human heart suggests that factors other than inhibition of PDE4 by milrinone may contribute to differences in cardiotonic action when compared to cilostazol. PMID- 12652112 TI - Improvement of aortic wall distensibility and reduction of oxidative stress by pioglitazone in pre-diabetic stage of Otsuka Long-Evans Tokushima fatty rats. AB - We tested the hypothesis that pioglitazone (insulin sensitizer) reduces oxidative stress and improves aortic wall distensibility in the pre-diabetic stage of Otsuka Long-Evans Tokushima Fatty rats, type 2 diabetes mellitus (DM) model. 20 DM and 9 nonDM male rats were divided into 3 groups: treated-DM, untreated-DM, and untreated-nonDM. Pioglitazone (0.01%) was mixed in chow in the treated group from 15 to 20 weeks of age. At baseline and 20 weeks, plasma malondialdehyde (MDA) was measured. At 20 weeks, intravascular ultrasound images and aortic pressure were simultaneously recorded. Stiffness parameter beta was calculated from the cyclic variations of aortic diameter and pressure. From an excised thoracic aorta, aortic wall collagen was measured, and the morphology was histopathologically evaluated by hematoxylin-eosin staining. At 20 weeks, MDA (nmol/ml) in treated-DM (2.3 +/- 0.3) was lower than in untreated-DM (3.2 +/- 0.6, p < 0.0001). beta in treated-DM (0.53 +/- 0.21) was smaller than that in untreated-DM (0.88 +/- 0.26, p = 0.0067). Aortic wall collagen (mg/100 mg dry weight) did not decrease in treated-DM (22.3 +/- 3.2 vs untreated-DM : 19.6 +/- 4.7). Lumen/medial area ratio (L/M) increased in treated-DM (2.79 +/- 0.40 vs untreated-DM : 2.22 +/- 0.20, p = 0.0041, untreated-nonDM : 2.25 +/- 0.55, p = 0.0075). MDA was significantly correlated with beta (r = 0.65, p = 0.0005) or L/M (r = -0.60, p = 0.0008). Pioglitazone may reduce oxidative stress and contribute to improvement of aortic wall stiffness without decrease in collagen content at an early prediabetic stage of type 2 DM. PMID- 12652113 TI - Effect of the calcium sensitizer levosimendan on the performance of ischaemic myocardium in anaesthetised pigs. AB - The calcium sensitizer levosimendan (LEV) improves the function of stunned myocardium, cardiac performance in heart failure, and possibly the efficiency of myocardial work. The present experiments investigated the effect of LEV on myocardial contraction and metabolism of acutely ischaemic myocardium distal to a functionally effective coronary artery stenosis. Anaesthetised open chest pigs (n = 14) were instrumented to assess heart rate (HR), aortic pressure (AoP), cardiac output (CO), blood flow in the left descending (QLAD) and circumflex (QLCX) coronary artery, myocardial end-diastolic segment length and systolic shortening (edL, MSS by sonomicrometry) in the LAD- and LCX-territory. Systemic vascular resistance (SVR), and a myocardial power index (PowI) for the LAD- and LCX-region were calculated. Following obstruction of QLAD by an external snare proximal to the first diagonal branch LEV was given intravenously (10 + 20 + 30 microg/kg 15 min apart, n = 8) or the vehicle of LEV (n = 6). Following LEV haemodynamics and regional myocardial performance changed significantly: HR +22 min(-1), AoP -6 mmHg, CO +17%, SVR -21%; intact myocardium: QLCX +15%, RLCX -24%, PowILCX + 39%; ischaemic myocardium: QLAD -7%, MSSLAD -42%, PowILAD -27%. The data confirm the pharmacological profile of LEV: positive chronotropy, positive inotropy, and vasodilatation. The pump function of acutely ischaemic myocardium worsened following LEV. The efficiency of myocardial performance did not improve. A beneficial effect of LEV on the function of ischaemic myocardium was possibly outmanoeuvred by the increase in heart rate. PMID- 12652115 TI - Use of cardiovascular drugs after acute myocardial infarction: a marked shift towards evidence-based drug therapy. AB - OBJECTIVE: To investigate the prescription pattern for cardiovascular drugs among patients discharged after an acute myocardial infarction (AMI) in hospitals that had participated in a corresponding study seven years earlier, and examine what the indications were for use of the different drugs. METHODS: From 16 hospitals we drew a sample of patients who were discharged with a diagnosis of AMI during a three months period in 1999/2000. Physicians in each hospital obtained from the medical records the observed rate of use of cardiovascular drugs at discharge. The drug use was compared with findings from a corresponding sample drawn in 1993. The main indication for use of the different cardiovascular drugs was recorded for the 1999/2000 sample. RESULTS: 399 patients discharged alive were included in the first study and 767 in the second. The use of beta-blockers, ACE inhibitors and statins rose substantially during the period. For patients aged 70 were: beta-blockers 45% vs 74%, ACE inhibitors 19% vs 38%, statins not recorded vs 35%. Aspirin/anticoagulant use was largely unchanged; 93% and 70% of patients aged 70 respectively used these drugs at the second survey. The use of regular nitrates and calcium antagonists had decreased. Nearly half of the >70 group and one-fifth of persons or =75) through the degree of benefit on hospital death rates associated with the use of thrombolytic therapy was attenuated after adjustment for additional confounders. CONCLUSIONS: Our findings indicate recent declines in the use of thrombolytic therapy in middle-aged and elderly patients with acute myocardial infarction. The impact of thrombolytic therapy on hospital outcomes was observed in each of our age strata under study though the magnitude of absolute and relative benefit varied according to age. PMID- 12652146 TI - Abciximab pharmacodynamics are unaffected by antecedent therapy with other GPIIb/IIIa antagonists in non-human primates. AB - BACKGROUND: Tirofiban and eptifibatide are currently approved for the medical stabilization of non-ST segment elevation acute coronary syndromes. In patients undergoing percutaneous coronary intervention (PCI) during infusion of these drugs, conversion to abciximab, which has long term proven clinical efficacy and cost-effectiveness, following PCI may be desirable. The purpose of this study was to determine if the binding or pharmacodynamics of abciximab is affected by a prior infusion of either tirofiban or eptifibatide. METHODS: In vitro binding experiments were performed to determine if prior exposure to tirofiban or eptifibatide altered the affinity and extent of binding of abciximab to GPIIb/IIIa. For in vivo experiments, cynomolgus monkeys were pretreated with a bolus and 18 hour infusion of saline, tirofiban, or eptifibatide. At the end of the initial treatment, a bolus and 12 hr infusion of abciximab was started without delay. Inhibition of platelet aggregation, GPIIb/IIIa receptor blockade and abciximab pharmacokinetics were measured during and after both infusions. RESULTS: Equilibrium binding of abciximab in vitro was unaffected by tirofiban or eptifibatide. The extent and duration of abciximab inhibition of ex vivo platelet aggregation, receptor blockade, and abciximab pharmacokinetics in monkeys during and after the abciximab infusion were not affected by prior infusion of the animals with tirofiban or eptifibatide. CONCLUSIONS: In vitro and in vivo studies revealed that the molecular interaction of abciximab with the platelet GPIIb/IIIa receptor is not altered by immediate prior exposure of platelets to small molecule GPIIb/IIIa antagonists. These preclinical studies suggest that the efficacy of abciximab should not be impaired if it is initiated following termination of therapy with small molecule GPIIb/IIIa antagonists. PMID- 12652147 TI - Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics. AB - BACKGROUND: We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described von Willebrand factor (vWF) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro. METHODS: Levels of hemostatic factors (vWF, plasminogen activator inhibitor type 1(PAI-1), antithrombin III (ATIII), in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique. In addition to comparing cell growth density and cell protein contents, cultured femoral arterial endothelial cells (FAECs) and cultured femoral venous endothelial cells (FVECs) were incubated with a series concentration of basic fibroblast factor (bFGF) (1, 10, 100 ng/ml) for up to 48 hours to test the amount of vWF secretion and morphological change. RESULTS: Both in tissue sections and cultured cells, the levels of vWF are higher in FVECs than in FAECs. We were unable to differentiate the level of PAI-1 and ATIII difference between FAECs and FVECs. bFGF (10 ng/ml) significantly increased vWF secretion from cultured FAECs but not from FVECs. The size of cultured FAECs is smaller than of FVECs; however, FAECs have higher amounts of protein contents than FVECs. CONCLUSIONS: These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs. These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease. PMID- 12652149 TI - Use of discharge-worksheet enhances compliance with evidence-based myocardial infarction care. AB - BACKGROUND: Prior studies demonstrate that effective secondary prevention therapies are underutilized in patients with myocardial infarction (MI) at hospital discharge. At a US tertiary center, we developed and encouraged providers to complete a simple "Acute MI Discharge Worksheet" (MIDW) designed to educate patients, prompt caregivers, and provide chart documentation regarding evidence-based therapies post-MI. METHODS AND RESULTS: The MIDW was introduced in May of 2000 with use encouraged in all surviving patients with MI. We calculated a patient discharge score by summing the number of quality indicators (aspirin use, beta-blocker use, ACE-inhibitor use, smoking cessation, lipid-lowering therapy, cardiac rehabilitation referral) and compared documentation of quality indicators at discharge between patients without (Group I, n = 65) and with (Group II, n = 60) the MIDW. Group II was subdivided into those with an incomplete worksheet (Group IIa, n = 26), and those with a completed worksheet (Group IIb, n = 34). Greater documentation of secondary prevention indicators occurred in patients with incomplete and completed discharge forms present. Mean Discharge scores were significantly higher for Group II vs. Group I (4.98 vs. 3.88, p < 0.0001), and Group IIb vs. Group IIa, (5.47 vs. 4.35, p < 0.001). CONCLUSION: A simple "Acute MI Discharge Worksheet" was associated with better adherence and documentation of evidence-based post MI care and be a useful component to improve post MI care. PMID- 12652148 TI - An automated strategy for bedside aPTT determination and unfractionated heparin infusion adjustment in acute coronary syndromes: insights from PARAGON A. AB - BACKGROUND: Intravenous unfractionated heparin remains a cornerstone of anticoagulation therapy for patients with acute coronary syndromes, but regulation to a target aPTT is challenging. We assessed unfractionated heparin infusion regulation by bedside, whole-blood aPTT testing and computerized, algorithmic infusion adjustment, and further evaluated the relationship of achieving the target aPTT with clinical outcomes. METHODS AND RESULTS: We studied 1,275 patients randomized to unfractionated heparin in PARAGON-A, which tested lamifiban with or without unfractionated heparin versus unfractionated heparin. All patients had baseline and 6-hour blinded, bedside aPTTs, then aPTTs per algorithm. A central computer translated encrypted values to algorithmic dose adjustment commands. We assessed the ability to achieve and maintain aPTTs of 50 70 seconds and associations of 6- and 12-hour aPTTs and time-to-target with 30 day outcomes.Overall, the median 6-hour aPTT was 50-70 seconds and remained so throughout infusion. Individually, only 33.6% of patients achieved 6-hour target range aPTTs, and only 40% of all aPTTs were in-range. After achieving target, only 42% of subsequent measures were in-range. Thirty-day death or myocardial infarction (death/MI) increased non-significantly as time-to-target increased (p = 0.08). Thirty-day mortality was similar if target aPTT was reached, regardless of timing. Death/MI trended lower if target aPTT was reached by 8 hours (p = 0.10). The best clinical outcomes were associated with in-range aPTTs. CONCLUSIONS: This study represents the most systematic monitoring and regulation of unfractionated heparin anticoagulation to date. Although average anticoagulation achieved target range, wide inter- and intra-patient variability may have important implications for clinical outcomes. PMID- 12652150 TI - Aspirin resistance and genetic polymorphisms. AB - Differences in genetic makeup or polymorphisms can affect individual drug response. Detecting genetic variation may help predict how a patient will respond to a drug and could be used as a tool to select optimal therapy, tailor dosage regimens, and improve clinical outcomes. The data are replete relative to the therapeutic efficacy of aspirin (ASA) for the prevention of ischemic events. However, there is a paucity of published data on the relationship between polymorphisms and the clinical effects on ASA. Prothrombotic genetic variations that may contribute to ASA resistance, and increased risk of cardiovascular events may involve: (1) a polymorphism on the cyclooxygenase-1 (COX-1) gene affecting Ser529; (2) overexpression of COX-2 mRNA on platelets and endothelial cells; (3) polymorphism PLA1/A2 of the gene encoding glycoprotein IIIa (GPIIIa); and (4) the homozygous 807T (873A) polymorphism allied with increased density of platelet GP Ia/IIa collagen-receptor gene. Because of the possible increased risk of ischemic vascular events, carriers of these genetic polymorphisms may be resistant to the antithrombotic effects of ASA and should be considered for additional or alternative treatment. PMID- 12652151 TI - Physician attitudes concerning anticoagulation services in the long-term care setting. AB - OBJECTIVES: There is little experience in the use of specialized anticoagulation services in the long-term care setting. Even less is known about physician attitudes regarding these services. To examine this issue, we surveyed physicians caring for nursing home residents in a sample of long-term care facilities located in Connecticut. METHODS: We surveyed physicians providing care to nursing home residents of a convenience sample of 21 Connecticut nursing homes. (These facilities had participated in a quality assessment and improvement project on preventing strokes in nursing home residents with atrial fibrillation.) Physicians were requested to complete a structured questionnaire about the challenges to managing nursing home residents on warfarin therapy and preferences concerning the use of an anticoagulation service to manage warfarin therapy in this setting. RESULTS: A total of 245 physicians were asked to participate in the survey, and 114 (47%) responded between November 5, 1999 and January 14, 2000. Of the 114 physicians who returned the survey, 91 reported that they currently cared for residents in long-term care facilities and thus completed the questionnaire. The majority of respondents agreed or strongly agreed that an anticoagulation service would reduce the workload on physicians, increase the costs of care for nursing home residents on warfarin, and increase the percent of time that nursing home residents on warfarin are maintained in the target therapeutic range. Most physicians disagreed or strongly disagreed with statements suggesting an anticoagulation service would decrease the costs of care for nursing home residents on warfarin, reduce the liability of the prescribing physician, interfere with their ability to care for patients on warfarin therapy, and reduce the risk of warfarin-related bleeding. Forty-five percent of respondents agreed with a statement that an anticoagulation service would intrude on physician decision-making. Only about half (53%) of the respondents indicated that they would or might utilize an anticoagulation service for managing their long-term care patients on warfarin. CONCLUSIONS: Use of a specialized anticoagulation service to manage warfarin therapy is a systems-level approach with the potential to improve the effectiveness and safety of this treatment. Physician skepticism regarding the usefulness of anticoagulation services will only be overcome by subjecting this approach to rigorous evaluation and by assuring physicians of their ongoing involvement in decision-making regarding warfarin therapy in their patients. PMID- 12652152 TI - Problems, interventions and complications in long-term oral anticoagulation therapy. AB - BACKGROUND: If problems, interventions and complications occurring during oral anticoagulation (OAC) are related with age, indication for OAC, hypertension, diabetes, previous stroke, and number of additional drugs. MATERIAL AND METHODS: Clinical characteristics, additional drugs, problems, interventions and complications of outpatients whose OAC was controlled between two years were registered. Potential gastrointestinal and urologic bleeding sources were eliminated prior to initiation of OAC. Five-hundred-seventy-nine patients (mean age 65 years, 44% female) were observed for 590 patient-years. RESULTS: Medical problems occurred in 352/100 patient-years (% p-y), organisational problems in 276% p-y, interventions in 636% p-y and complications in 13.8% p-y. Patients >65 years had less organisational problems (254 vs. 302% p-y, p = 0.0092) and interventions (574 vs. 713% p-y, p = 0.0003) than patients < or =65 years. The 35 patients with heart valve prosthesis had more life-threatening and fatal complications (12% p-y) than the 360 patients with atrial fibrillation (1.0% p y), 128 patients with venous thromboembolism or 56 patients with other indications (0.0% p-y, p = 0.0024). Problems, interventions and complications were not related with hypertension (n = 297), diabetes (n = 97) or previous stroke (n = 90). Patients with >3 additional drugs/day had a higher complication rate than patients with < or =3 drugs/day (21 vs. 8.7% p-y, p = 0.0238). Patients with complications had more headache (27 vs. 20% p-y, p = 0.0036), chest pain (45 vs. 27% p-y, p = 0.0150), abdominal pain (25 vs. 15% p-y, p = 0.0350) and pain in the limbs (55 vs. 42% p-y, p = 0.0044) than patients without complications. CONCLUSIONS: By careful monitoring, eliminating potential bleeding sources, treating pain adequately and minimizing additional drugs the complications of OAC can be kept low. PMID- 12652153 TI - Association of changes in D-dimer and other coagulation markers with changes in Marder score after treatment of acute venous thrombosis. AB - AIM: Coagulation markers are sensitive tools to assess ongoing thrombus formation. An association between changes in these markers and changes in venographic Marder scores in patients with acute deep vein-thrombosis treated with low-molecular-weight (LMWH) or unfractionated heparin (UFH) has not been reported. METHODS: We investigated differences in coagulation parameters before and at the end of a twelve days the treatment of patients with an improvement versus no improvement of the venographic findings at the end of the treatment with LMWH (n = 48) and UFH (n = 41). RESULTS: Patients with lower values in the Marder score had lower D-dimer levels at day 12 compared to entry treated with UFH and LMWH (p < 0.001). Not improved Marder scores paralleled unchanged D-dimer levels at end of both treatments. Higher values of factor-Xa inhibition and Heptest assay (p < 0.001) were measured at the end of treatment in LMWH- in contrast to UFH-patients. Thrombin inhibition was lower and unchanged at day 12 in patients treated with LMWH and UFH, respectively. Thrombin generation inhibition and release of tissue-factor pathway inhibitor remained unchanged in both groups. CONCLUSION: An improved Marder score is associated with a decrease of D-dimer during UFH and LMWH treatment of deep vein-thrombosis. PMID- 12652154 TI - Fatal pulmonary artery embolism in a sickle cell patient: case report and literature review. AB - This article reports a rare case of fatal saddle embolism to the pulmonary artery presenting as an Acute Chest Syndrome (ACS) in a Sickle Cell patient. We present a review of the etiology, pathophysiology, clinical manifestation and management of ACS. Clinicians should be aware of the fact that a sickle cell crisis admission may represent a life-threatening process. Such awareness will help physicians to act promptly and execute proper therapeutic interventions. It is important for clinicians to be expectant of impending clinical deterioration and likewise be aware that ACS can develop in patients hospitalized for other medical or surgical conditions. PMID- 12652155 TI - The role of the NO axis and its therapeutic implications in pulmonary arterial hypertension. AB - Pulmonary Arterial Hypertension (PAH) is a disease of the pulmonary vasculature leading to vasoconstriction and remodeling of the pulmonary arteries. The resulting increase in the right ventricular afterload leads to right ventricular failure and death. The treatment options are limited, expensive and associated with significant side effects. The nitric oxide (NO) pathway in the pulmonary circulation provides several targets for the development of new therapies for this disease. However, the NO pathway is modulated at multiple levels including transcription and expression of the NO synthase gene, regulation of the NO synthase activity, regulation of the production of cyclic guanomonophosphate (cGMP) by phosphodiesterases, postsynthetic oxidation of NO, etc. This makes the study of the role of the NO pathway very difficult, unless one uses multiple complementary techniques. Furthermore, there are significant differences between the pulmonary and the systemic circulation which make extrapolation of data from one circulation to the other very difficult. In addition, the role of NO in the development of pulmonary hypertension varies among different models of the disease. This paper reviews the role of the NO pathway in both the healthy and diseased pulmonary circulation and in several animal models and human forms of the disease. It focuses on the role of recent therapies that target the NO pathway, including L-Arginine, inhaled NO, the phosphodiesterase inhibitor sildenafil and gene therapy. PMID- 12652156 TI - Role of nitric oxide in matrix remodeling in diabetes and heart failure. AB - Accumulation of oxidized-matrix between the endothelium and myocytes is associated with endocardial endothelial (EE) dysfunction in diabetes and heart failure. High levels of circulating homocysteine (Hcy) have been demonstrated in diabetes mellitus (DM). These high levels of Hcy (hyperhomocysteinemia, HHcy) have a negative correlation with peroxisome proliferator activated receptor (PPAR) expression. Studies have demonstrated that Hcy decreases bioavailability of endothelial nitric oxide (eNO), generates nitrotyrosine, and activates latent matrix metalloproteinase (MMP), instigating EE dysfunction. PPAR ligands ameliorate endothelial dysfunction and DM. In addition Hcy competes with PPAR ligands. The understanding of molecular, cellular, and extracellular mechanisms by which Hcy amplifies DM will have therapeutic ramifications for diabetic cardiomyopathy. PMID- 12652157 TI - Nitric oxide and cardiovascular protection. AB - Nitric oxide (NO) plays a critical role in ischemic heart disease and ischemia reperfusion. There is an increasing body of evidence to support the role of NO in myocardial and vascular protection in disease. The finding that NO might act as a trigger of late ischemic preconditioning (IPC) might lead to the development of novel anti-ischemic therapy. The role of NO signaling in the cardioprotective effects of ACE inhibitors and angiotensin II type 1 receptor(AT(1)) receptor antagonists is an active area of study. PMID- 12652158 TI - Role of nitric oxide in the pathophysiology of heart failure. AB - Nitric oxide (NO) plays critical roles in the regulation of integrated cardiac and vascular function and homeostasis. An understanding of the physiologic role and relative contribution of the three NO synthase isoforms (neuronal--NOS1, inducible--NOS2, and endothelial--NOS3) is imperative to comprehend derangements of the NO signaling pathway in the failing cardiovascular system. Several theories of NO and its regulation have developed as explanations for the divergent observations from studies in health and disease states. Here we review the physiologic and pathophysiologic influence of NO on cardiac function, in a framework that considers several theories of altered NO signaling in heart failure. We discuss the notion of spatial compartmentalization of NO signaling within the myocyte in an effort to reconcile many controversies about derangements in the influences of NO in the heart and vasculature. PMID- 12652159 TI - Nitric oxide, platelet function, myocardial infarction and reperfusion therapies. AB - Platelets play an important role in physiologic hemostasis and pathologic thrombosis that complicate the course of vascular disorders. A number of platelet functions including adhesion, aggregation and recruitment are controlled by nitric oxide (NO) generated by platelets and the endothelial cells. Derangements in this generation may contribute to the pathogenesis of thrombotic complications of vascular disorders. The pharmacologic supplementation of the diseased vasculature with drugs releasing NO may help to restore the hemostatic balance. PMID- 12652160 TI - The therapeutic effect of natriuretic peptides in heart failure; differential regulation of endothelial and inducible nitric oxide synthases. AB - The abnormal regulation of nitric oxide synthase activity represents an underlying feature of heart failure. Increased peripheral vascular resistance, and decreased renal function may be in part related to impaired endothelium dependent nitric oxide (NO) synthesis. Paradoxically, the chronic production of NO by inducible nitric oxide synthase (iNOS) in heart failure exerts deleterious effects on ventricular contractility, and circulatory function. Consequently, pharmacologically improving endothelium-dependent NO synthesis and the concomitant inhibition of iNOS activity would be therapeutically advantageous. Interestingly, natriuretic peptides have been shown to differentially regulate endothelial NOS (eNOS) and iNOS activity. Moreover, in both patients and animal models of heart failure, pharmacologically increasing plasma natriuretic peptide levels ameliorated vascular tone, renal function, and ventricular contractility. Based on these observations, the following review will explore whether the therapeutic benefit of the natriuretic peptide system in heart failure may occur in part via the amelioration of endothelium-dependent NO synthesis, and the concomitant inhibition of cytokine-mediated iNOS expression. PMID- 12652163 TI - Statins and the role of nitric oxide in chronic heart failure. AB - Endothelial dysfunction plays an important role in a number of cardiovascular diseases. An important pathogenetic factor for the development of endothelial dysfunction is lack of nitric oxide (NO), which is a potent endothelium-derived vasodilating substance. 3-Hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors (statins), originally designed to lower plasma cholesterol levels, seem to ameliorate endothelial dysfunction by a mechanism so far only partly understood. However, statins increase nitric oxide synthase activity. It has been speculated that this and other "side effects" of statin treatment are due to inhibition of Rho, an intracellular signalling protein that initiates Rho kinase transcription. Moreover, statins possess anti-inflammatory characteristics. Some statins have proven to lower plasma levels of C-reactive protein, which is induced by pro-inflammatory cytokines. Other statins have been demonstrated to directly inhibit pro-inflammatory cytokine induction. Finally, some data suggest that statins might be able to counterbalance an increased production of oxygen free radicals. Since chronic heart failure is accompanied not only by endothelial dysfunction, but also by pro-inflammatory cytokine activation and enhanced formation of oxygen free radicals, it is tempting to speculate that statins might be an ideal candidate to treat certain features of this disease. The doses needed to achieve the desired effects might be much lower than those needed to treat hypercholesterolemia. PMID- 12652161 TI - Nitric oxide, atherosclerosis and the clinical relevance of endothelial dysfunction. AB - The endothelium plays a key role in vascular homeostasis through the release of a variety of autocrine and paracrine substances, the best characterized being nitric oxide. A healthy endothelium acts to prevent atherosclerosis development and its complications through a complex and favorable effect on vasomotion, platelet and leukocyte adhesion and plaque stabilization. The assessment of endothelial function in humans has generally involved the description of vasomotor responses, but more widely includes physiological, biochemical and genetic markers that characterize the interaction of the endothelium with platelets, leukocytes and the coagulation system. Stable markers of inflammation such as high sensitivity C-reactive protein are indirect and potentially useful measures of endothelial function for example. Attenuation of the effect of nitric oxide accounts for the majority of what is described as endothelial dysfunction. This occurs in response to atherosclerosis or its risk factors. Much remains to be learned about the molecular and genetic pathophysiological mechanisms of endothelial cell abnormalities. However, pharmacological intervention with a growing list of medications can favorably modify endothelial function, paralleling beneficial effects on cardiovascular morbidity and mortality. In addition, several small studies have provided tantalizing evidence that measures of endothelial health might provide prognostic information about an individual patient's risk of subsequent events. As such, the sum of this evidence makes the clinical assessment of endothelial function an attractive surrogate marker of atherosclerosis disease activity. The review will focus on the role of nitric oxide in atherosclerosis and the clinical relevance of these findings. PMID- 12652165 TI - Assessing the communication skills of doctors in training: reliability and sources of error. AB - Medical examining bodies now commonly assess candidates' communication skills. However, there are a number of within-case sources of error that can mean examinations have poor reliability and validity. The aims of this study were to determine the main within-case sources of error and to identify the best methods of maximising reliability in highly structured communication skills assessments. Subjects were fifth year medical students at the University of Aberdeen. Subjects were videotaped giving clinical information to standardised patients and relatives toward the end of their fifth year and toward the end of their pre registration year. Sources of variation, reliability coefficients and the most effective methods of reducing measurement error were calculated using generalisability theory. Systematic differences among subjects' communication skill was the main source of variation in two of the three assessment cases. The implications of the results are discussed. PMID- 12652167 TI - Quality assurance methods for performance-based assessments. AB - Performance assessments are subject to many potential error sources. For performance-based assessments, including standardized patient (SP) examinations, these error sources, if left unchecked, can compromise the validity and reliability of scores. Quality assurance (QA) measures, both quantitative and qualitative, can be used to ensure that candidate scores are accurate and reasonably free from measurement error. The purpose of this paper is to outline several QA strategies that can be used to identify potential content- and score related problems with SP assessments. These approaches include case analyses and various comparisons of primary and observer scores. Specific examples from the ECFMG Clinical Skills Assessment (CSA) are used to educate the reader concerning appropriate statistical methods and legitimate data interpretations. The results presented in this investigation highlight the need for well-defined training regimes, regular feedback to those involved in rating/scoring performances, and detailed statistical analyses of all scores. PMID- 12652164 TI - The nitric oxide-endothelin-1 connection. AB - Nitric oxide (NO) and endothelin-1 (ET-1) are endothelium-derived mediators that play important roles in vascular homeostasis. This review is focused on the role and reciprocal interactions between NO and ET-1 in health and diseases associated with endothelium dysfunction. We will also discuss the clinical significance of NO donors and drugs that antagonize ET receptors. PMID- 12652162 TI - Acute and chronic endothelial dysfunction: implications for the development of heart failure. AB - Heart failure has been characterized by a reduction in cardiac contractile function resulting in reduced cardiac output. The clinical symptoms including mild tachycardia, reduced arterial pressure, increased venous or filling pressure and exercise intolerance have conceptually, to a large degree, been attributed to cardiac myocyte dysfunction. More recently, a vascular component has been recognized to contribute to heart failure. Among the most studied vascular mechanisms that might contribute to the development of heart failure has been the reduced production of nitric oxide or the reduced bioactivity of NO associated with both basic models of heart failure and disease in patients. The still evolving concept that heart failure is a cytokine activated state has, in addition, focused attention on the possibility that the cytokine driven isoform of NO synthase (NOS), iNOS, may produce sufficient quantities of NO to actually suppress cardiac myocyte function contributing to the reduced inotropic state in the failing heart. Thus, our view of the role of NO in the development of heart failure has evolved from simply a reduction in production of NO in blood vessels, to altered substrate availability (i.e. L-arginine), to increased scavenging of NO by superoxide anion, to increased production of NO from iNOS. As these concepts develop, our approach to the therapeutics of heart failure has also progressed with the recognition of the need to develop treatments directed towards addressing one or more of these etiologies. This review will focus on these aspects of the involvement of NO in the development of heart failure and some of the treatments that have developed from our understanding of the basic biology of NO to address these pathohysiologic states. PMID- 12652166 TI - Practice makes perfect: the critical role of mixed practice in the acquisition of ECG interpretation skills. AB - PURPOSE: To examine the effect of instructional format on medical students' learning of ECG diagnosis. METHOD: Two experiments employed different learning and practice methods. In the first, students were randomly allocated to one of two instructional approaches, one organized around features (e.g., QRS voltage) and the other around diagnostic categories (e.g., bundle branch blocks), followed by a practice phase. In the second experiment, the instruction was standardized, and students were randomly allocated to one of two practice phases, either "contrastive" where examples from various categories are mixed together, or "non contrastive" where all the examples in a single category are practiced in a single block. RESULTS: In the first experiment, there was no significant differences in students' diagnostic accuracy on novel ECG examples. In the second experiment, students exposed to the contrastive approach in the practice phase had superior diagnostic accuracy (46%) compared to 30% accuracy for the non contrastive session, p < 0.05). CONCLUSION: These experiments highlight two important features in the design of instructional materials. First, learning around the features of the problem (analogous to problem-based learning) may have no advantages over learning the category. Second, the design and organization of deliberate practice can result in significant learning gain. PMID- 12652168 TI - What does "The discovery of grounded theory" have to say to medical education? PMID- 12652169 TI - Charles W. Dohner, PhD: an evaluator and mentor in medical education. AB - As one of the first generation medical education pioneers, Charles W. Dohner, PhD established the ninth office of medical education at the University of Washington (UW) where he served as chairman from 1967-1996. With a background in education and measurement, he focused his work on evaluation of educational programs and faculty development. The Department of Medical Education went through three distinct stages of development: pathfinding 1967-1972 focused on developing working relationships with the faculty and clarifying identity, integration into academic affairs 1972-1980, and direct leadership by department faculty 1980 1996. Dohner helped to create and evaluate the WAMI program, a regional medical education program for the states of Washington, Alaska, Montana, and Idaho. He served as a consultant to a specialty board, the founding president of the Society of Directors of Research in Medical Education, and a frequent consultant in international medical education. Dohner identified three important innovations in medical education: educators in academic medicine, simulations and performance assessment, and community-based medical education. Success factors for professional education include technical competence in education, interpersonal communication and collaboration skills, a plan for personal growth, and use of mentors. Future trends in medical education will involve information technology, professionalism, wellness and complementary medicine, and performance assessment. He has been a passionate spokesman for excellence in medical education and most noted for his roles as an evaluator, program developer, and mentor of academic leaders. PMID- 12652170 TI - Impartial judgment by the "gatekeepers" of science: fallibility and accountability in the peer review process. AB - High publication demands and the low acceptance rate of peer review journals place the journal editors and their reviewers in a powerful position. Journal reviewers have a vital role not only in influencing the journal editor's publication decisions, but also in the very nature and direction of scientific research. Because of their influence in peer review outcomes, journal reviewers are aptly described as the "gatekeepers of science." In this article we describe several pitfalls that can impede reviewers' impartial judgement. These include such issues as confirmatory bias, the negative results bias (the file drawer problem), the Matthew effect, the Doctor Fox effect, and gender, race, theoretical orientation, and "political correctness." We argue that procedures currently used by many professional journals, such as blind or masked review, may not completely alleviate the effects of these pitfalls. Instead, we suggest that increasing reviewers' awareness of the pitfalls, accountability, and vigilance can improve fairness in the peer review process. The ultimate responsibilities belong to the journal editors who are confronted with the difficult task of satisfying journal readers, contributors, reviewers, and owners. We recommend that the journal editors conduct periodic internal and external evaluations of their journals' peer review process and outcomes, with participation of reviewers, contributors, readers and owners. PMID- 12652171 TI - Classifications and practice guidelines in atrial fibrillation: a changing landscape. PMID- 12652172 TI - Identification of transmural necrosis along a linear catheter ablation lesion during atrial fibrillation and sinus rhythm. AB - BACKGROUND: Determining whether a linear catheter radio frequency (RF) ablation lesion is transmural may be difficult, especially during atrial fibrillation. We hypothesized that changes in pacing thresholds and electrogram amplitude during atrial fibrillation and sinus rhythm could be used to assess whether a radiofrequency ablation resulted in transmural necrosis. METHODS: A hexapolar, linear, RF ablation catheter was positioned between the caval veins in the right atrium of seven sheep. Pacing thresholds and electrogram amplitudes during atrial fibrillation and sinus rhythm were measured before and after the application of RF energy. Sites along the linear lesion were assessed histologically. RESULTS: The electrogram amplitude in atrial fibrillation decreased significantly more at transmural sites (unipolar recording: 33 +/- 11% transmural vs. 22 +/- 13% non transmural, p < or = 0.01; bipolar recording: 62 +/- 9% transmural vs. 43 +/- 15% non-transmural, p < or = 0.01). The electrogram amplitude in sinus rhythm decreased significantly more at transmural sites (unipolar recording: 49 +/- 18% transmural vs. 15 +/- 20% non-transmural, p < 0.001; bipolar recording: 63 +/- 17% transmural vs. 42 +/- 19% non-transmural, p = 0.002). The pacing threshold increased significantly more at sites with transmural necrosis (unipolar: increased by 378 +/- 103% transmural vs. 207 +/- 93% non-transmural, p < 0.001; bipolar: 370 +/- 80% transmural vs. 259 +/- 60% non-transmural, p < 0.001). CONCLUSIONS: The amplitude of the atrial electrogram from an ablation catheter can be used to discriminate areas with transmural necrosis from those without transmural necrosis during either atrial fibrillation or sinus rhythm. Termination of atrial fibrillation may not be necessary to estimate the histologic characteristics of an ablation lesion. PMID- 12652173 TI - Pharmacological cardioversion of recent onset atrial fibrillation with intravenous amiodarone in patients receiving long-term amiodarone therapy: is it reasonable? AB - In clinical practice the use of intravenous amiodarone has been proposed for the conversion of recurrent atrial fibrillation in patients already under chronic treatment with the same drug. Given that intravenous amiodarone exhibits different electrophysiological properties than when the drug is taken orally over a long period, this approach seems reasonable, but its effectiveness and safety have not been investigated systematically before. Of 45 patients under chronic treatment with amiodarone for the maintenance of sinus rhythm who had atrial fibrillation of recent onset, 23 were given intravenous loading of the same drug for 24 hours and 22 received placebo. Nine patients underwent an electrophysiological study several months after the successful restoration of sinus rhythm, before and after another intravenous loading dose of amiodarone, in order to examine the possible electrophysiological changes. In the amiodarone group 20 patients were successfully converted to sinus rhythm, compared to 13 of the placebo group (p < 0.05). No serious side effects of the intravenous administration were observed. Prolongation of refractoriness was seen in all 9 patients who underwent electrophysiological study after intravenous loading, without any effect on repolarization, atrioventricular conduction or sinus node function. In conclusion an intravenous loading dose of amiodarone exerts an additional electrophysiological effect in patients already under chronic treatment with the same drug. Such a combined therapy could be used with a high efficacy and safety for the conversion of recent onset atrial fibrillation in patients who are receiving long-term amiodarone therapy. PMID- 12652175 TI - Dynamic alterations in right atrial activation during atrial fibrillation. AB - Atrial fibrillation (AF) is usually considered to be a disorganized rhythm. However, several reports have described areas of organized right atrial (RA) activation during AF. We describe a patient with focal paroxysmal AF with a consistent pattern of organized RA activity despite typically disorganized left atrial activity. The sequence of RA activation varied, depending on which pulmonary vein foci initiated the AF. PMID- 12652174 TI - Real-time cardiac catheter navigation on three-dimensional CT images. AB - INTRODUCTION: Targets for ablation of atrial fibrillation, atrial flutter, and non-idiopathic ventricular tachycardia are increasingly being selected based on anatomic considerations. Because fluoroscopy provides only limited information about the relationship between catheter positions and cardiac structures, and is associated with radiation risk, other approaches to mapping may be beneficial. METHODS: The spatial and temporal information of an electromagnetic catheter tip position sensing system (Magellan, Biosense Inc.) was superimposed on a three dimensional (3D) CT of the chest in swine using fiducial markers for image registration. Position and orientation of a 6 French catheter with an electromagnetic sensor was displayed in real-time on a corresponding 3D-CT. Catheter navigation within the heart and the great vessels was guided by detailed knowledge about catheter location in relation to cardiac anatomy. RESULTS: Anatomic structures including the atrial septum, pulmonary veins, and valvular apparatus were easily identified and used to direct catheter navigation. During the right heart examination, the catheter was navigated through the superior and inferior vena cava to predetermined anatomic locations in right atrium, right ventricle and pulmonary artery. The ablation catheter was also navigated successfully from the aorta through the aortic valve in the left ventricle. No complication was encountered during the experiments. The accuracy and precision of this novel approach to mapping was 4.69 +/- 1.70 mm and 2.22 +/- 0.69 mm, respectively. CONCLUSIONS: Real-time display of catheter position and orientation on 3D-CT scans allows accurate and precise catheter navigation in the heart. The detailed anatomic information may improve anatomically based procedures like pulmonary vein ablation and has the potential to decrease radiation times. PMID- 12652176 TI - Monophasic action potential-like electrocardiogram simulating acute myocardial infarction. AB - Monophasic action potential-like electrocardiogram simulating acute myocardial infarction has been reported in patients with hyperkalemia. We report two cases presenting marked diffuse RS-T elevation resembling monophasic action potential. They had abnormal electrolyte levels, including hyperkalemia and hypocalcemia in case 1 and hypokalemia and hypocalcemia in case 2. The ECG abnormality returned to baseline after correction of potassium and calcium. We conclude that monophasic action potential-like electrocardiogram is caused by electrolyte disturbance other than acute myocardial infarction. PMID- 12652177 TI - Impact of automatic adjustment of stimulation outputs on pacemaker longevity in a new dual-chamber pacing system. AB - INTRODUCTION: Automatic capture verification using the Autocapture (AC) feature enabled by paced evoked response detection and delivery of high energy back-up pulses intends to increase patient safety. Furthermore, adjustment of stimulation outputs can save energy and potentially improve pacemaker (PM) longevity. The purpose of this study was to evaluate the theoretical longevity of a new dual chamber PM with the integrated AC feature (Affinity DR, St. Jude Medical) in comparison to the longevity of a previous model from the same manufacturer without AC (Trilogy DR). METHOD: Affinity PMs were implanted in 16 patients and connected to a compatible lead with low polarization properties. AC was activated when the evoked response was significantly higher than the polarization voltage. Theoretical PM longevity was calculated with and without AC during follow-up. The measured and calculated values were compared to measurements in 19 patients, who consecutively received Trilogy-PMs during the same time period. RESULTS: In only one patient the evoked response was not adequate, and as a result, AC was not programmed. The calculated longevity of the Affinity-PMs 79 +/- 28 days after implantation was significantly higher in comparison to the Trilogy-PMs (Affinity PMs: 8.9 +/- 1.2 years without and 9.5 +/- 1.1 years with AC; Trilogy-PMs: 6.5 +/ 0.8 years) (p < 0.005). CONCLUSION: The AC feature is an optional algorithm that can be activated in most patients and it significantly prolongs predicted battery longevity due to automatic adjustment of stimulation outputs. PMID- 12652179 TI - Atrial fibrillation ablation leads to long-term improvement of quality of life and reduced utilization of healthcare resources. AB - In some patients, rapid activation from one or several foci can lead to atrial fibrillation. This study evaluated long-term changes in quality of life and healthcare resource utilization in patients with atrial fibrillation treated by ablation of focal triggers. Thirty-three patients underwent ablation for paroxysmal atrial fibrillation. Health surveys (SF-36) were obtained at baseline, and after 1 year and 3 years of follow-up. Health care costs were measured for the 3 years before and after ablation. Ablation was successful in 82%, partially successful in 12% (no sustained episodes but on antiarrhythmic drug therapy), and unsuccessful in 6% of patients. The average number of ablation procedures was 1.6 +/- 0.6 per patient. After ablation, patients reported significantly improved quality of life in all SF-36 categories except bodily pain. Healthcare resource utilization was significantly reduced after ablation (Clinic visits: 7.4 +/- 2.5 per year vs. 1.1 +/- 0.6 per year, p < 0.05; Emergency room visits: 1.7 +/- 0.90 per year vs. 0.03 +/- 0.17 per year, p < 0.05; Hospitalization: 1.6 +/- 0.81 vs. 0, p < 0.05). Cost of healthcare (not including procedural costs) was significantly reduced after ablation (Pre-ablation: 1,920 +/- 889 dollars/year vs. post-ablation: 87 +/- 68 dollars/year; p < 0.01). Procedural cost of ablation was 17,173 +/- 2,466 dollars/patient. Ablation of focal triggers of atrial fibrillation is associated with a sustained improvement in quality of life. Although the initial cost of ablation is high, after ablation, utilization of healthcare resources is significantly reduced. PMID- 12652178 TI - Selective atrionodal input ablation for induction of proximal complete heart block with stable junctional escape rhythm in patients with uncontrolled atrial fibrillation. AB - BACKGROUND: The study tests the hypothesis that ablating all inputs to the atrioventricular (AV) node can result in complete heart block with stable junctional escape rhythm. METHODS AND RESULTS: We attempted atrionodal input ablation in 76 consecutive patients with uncontrolled atrial fibrillation. Fast and slow pathways were first ablated. If there was no AV block, additional energy applications were done between fast and slow pathway locations. The patients were followed for 42 +/- 11 months. Group I (n = 57) comprised patients with complete heart block and junctional escape rhythm (53 +/- 4 beats/min) at the end of the procedure. The escape rhythm remained stable throughout follow-up. Group II (n = 15) were patients who failed the stepwise atrionodal input ablation and required AV junctional ablation guided by His bundle potential to achieve complete heart block. Four patients showed a slow escape rhythm after ablation (33 +/- 4 beats/min). Others had no escape rhythm. All 15 pts remained pacemaker dependent. The total death rate of groups I and II was 18/57 (31.6%) vs 10/15 (66.7%), respectively (p < 0.02). These differences could not be explained by a difference of left ventricular ejection fraction (0.42 +/- 0.07 vs 0.41 +/- 0.04, respectively, p = NS). CONCLUSIONS: (1) In most patients, ablation of both fast and slow pathways did not result in complete heart block, indicating the presence of multiple atrionodal inputs. (2) Ablation of all atrionodal inputs may result in complete heart block with stable junctional escape rhythm. (3) As compared with AV junctional ablation, atrionodal input ablation was associated with a lower mortality rate on long-term follow up. PMID- 12652180 TI - Comparison of coronary venous defibrillation with conventional transvenous internal defibrillation in man. AB - OBJECTIVE: Animal studies have shown that defibrillation in coronary veins is more effective than in the right ventricle. We aimed to assess the feasibility of placing defibrillation electrodes in the middle cardiac vein (MCV) in man and its impact on defibrillation requirements. METHODS: A prospective randomised study conducted in a tertiary referral centre. 10 patients (9 male) undergoing ICD implantation (65 (12) yrs) for NASPE/BPEG indications were studied. Defibrillation thresholds (DFT) were measured, using a binary search and an external defibrillator after 10 seconds of ventricular fibrillation, for the following configurations in each patient (order of testing randomised): RV + MCV -> Can and RV --> SVC + Can. INTERVENTIONS: A dual coil defibrillation electrode was placed transvenously in the right ventricle (RV) in the conventional manner. Using a guiding catheter a 3.2 Fr (67.5 mm length) electrode was placed transvenously in MCV. A test-can was placed subcutaneously in the left pectoral region. RESULTS: Lead placement was possible in 8/10 pts. Time to perform a middle cardiac venogram and place the electrode was 21 (23) mins. No adverse events were observed. Defibrillation current was less (6.7 (2.7) A) with RV + MCV --> Can compared to the conventional RV --> SVC + Can configuration (8.9 (3.4) A, p = 0.03). There was no significant difference in defibrillation voltage or energy. However, shock impedance was higher in the former configuration (57 (10) v. 43 (6) Omega, p = 0.001). CONCLUSIONS: In the majority of cases placement of a defibrillation lead in MCV is feasible. Defibrillation current requirements are 25% less when the shock is delivered using a MCV electrode. PMID- 12652181 TI - Successful implantation of cardiac defibrillators without induction of ventricular fibrillation using upper limit of vulnerability testing. AB - INTRODUCTION: Conventionally, the implantable cardioverter-defibrillator (ICD) is tested at implantation by measurement of defibrillation threshold (DFT), which involves repeated induction of ventricular fibrillation (VF). We report our data on successful ICD implantation without VF induction using a modified upper limit of vulnerability (ULV) testing method, compared to standard DFT testing. METHODS: Fourteen patients underwent ICD implantation using a modified ULV testing method by delivering a 15 J shock during the vulnerable period on the peak of the T wave, and if VF was not induced 15 J shocks were repeated at -20 and -40 msec before the peak of T wave. Failure to induce VF, indicating a ULV <15 joules (J), suggested a DFT < or =20 J based on previous studies demonstrating a close correlation (+/-5 J) between ULV and DFT. If VF was induced, a 20 J rescue shock was delivered. ICD therapy was then programmed on the basis of ULV testing. All patients underwent pre-discharge DFT testing to confirm adequate DFT. RESULTS: Using a modified ULV testing method, ICD implantation was completed without induction of VF in 8 patients and only a single episode of VF in 6 patients. The mean number of VF episodes (0.42 +/- 0.5) induced with ULV testing was significantly lower (p <.001) than the number induced during DFT testing (3.9 +/- 0.8). Pre-discharge DFT testing did not alter ICD programming in any patient. During follow-up of 14.85 +/- 12.31 months, three patients had seven episodes of VT/VF, six of whom were converted with the programmed first-shock strength, while one required a second high-energy shock to convert. This patient had a pre discharge DFT of 10 joules. CONCLUSIONS: Successful ICD implantation can be safely performed with no or fewer episodes of VF induction using a modified ULV testing method. PMID- 12652182 TI - Karel Frederik Wenckebach (1868-1940). PMID- 12652183 TI - Insulin-like growth factor 1 (IGF-1) and aging: controversies and new insights. AB - Insulin/insulin-like growth factor (IGF) signaling plays a major role in the control of aging and life span in invertebrates. Major extension of life span in growth hormone receptor knock out (GHR-KO) mice that are GH resistant, and subsequently, IGF-I-deficient indicates that similar mechanisms may operate in mammals. This conclusion is supported by association of reduced IGF-I levels and delayed aging in three different GH-deficient mutant mice and in animals subjected to caloric restriction, but is difficult to reconcile with neuroprotective effects of IGF-I and with the suspected role of declining GH levels during aging. We suggest that the role of IGF in the regulation of growth and adult body size is important in mediating the effects of longevity genes on aging and life span. Suspected mechanisms of IGF-I action in aging also include reduced insulin signaling, enhanced sensitivity to insulin, and reduced thermogenesis with diminished oxidative damage of macromolecules being the likely final common pathway of these effects. We suspect that IGF-I is important in evolutionarily conserved mechanisms that link life history, including development, reproduction, and aging with availability of energy resources. PMID- 12652184 TI - Effects of X-irradiation in early ontogenesis on the longevity and amount of the S1 nuclease-sensitive DNA sites in adult Drosophila melanogaster. AB - The long-term consequences of the X-irradiation of Drosophila melanogaster fruit fly one-hour eggs with doses of 0.25, 0.50, 0.75, 1, 2 and 4 Gy were investigated. Longevity hormesis was observed in males exposed to 0.5 Gy and 0.75 Gy, but no longevity increase was observed in females. The electrophoretic analysis has shown that the amount of the DNA segments resulting from cleavage in S1 nuclease-sensitive sites (<3 kb) reached 39.2% of the total DNA from control males. DNA from the irradiated males had a smaller amount of such fragments (10 30% in different experimental groups). These findings indicate that the longevity hormesis may be associated with irradiation-induced long-term structural and/or functional DNA modifications. PMID- 12652185 TI - Establishment of the mimetic aging effect in mice caused by D-galactose. AB - We successfully established an oxidant damage of mimetic aging model using mice induced by D-galactose, and the mimetic aging model is relative to free radical and the accumulation of waste substances in metabolism. The animals were divided into 3 groups: (1) phosphate-buffered saline (PBS); (2) 1% D-galactose; (3) 5% D galactose by subcutaneous injection every day. After 45 days, mice treated with D galactose showed a significant increase in the malondialdehyde (MDA), total antioxidant status (TAS) and a decrease in superoxide dismutase (SOD) in the blood compared with the PBS group. In the brain, the D-galactose treated mice exhibited a higher level MDA and a lower level SOD activity. In the liver, only the 5% D-galactose group indicated a significant increase in MDA. By reference to the oxidative biomarkers in blood, brain and liver, we have confirmed the reliability of the mimetic aging model. PMID- 12652186 TI - Mitochondrial electron transport chain complexes in aging rat brain and lymphocytes. AB - Several lines of evidence suggest that mitochondria not only plays a key role in the causation of aging, but it is also the most adversely affected organelle during aging. The aim of the present study was to examine the role of mitochondria in the aging process by determining whether the activities of various 'electron transport chain' (ETC) complexes are deleteriously affected during aging in the rat brain and lymphocytes. The enzyme activity of ETC complexes I-V, and citrate synthase (CS) were assayed from discrete brain areas viz., cerebral hemisphere (CH), cerebellum (CB), brain stem (BS) and diencephalon (DC), along with the lymphocytes from four different age groups of rats, namely, 1-month-old rats, 3-4-month-old adults, 12-month-old adults and 24-month-old aged rats. Significant decline was observed in all the enzyme activities in 12 and 24 month-old rats as compared to 3-4-month-old adult rats. Moreover, one-month-old rats showed lower levels of ETC complexes as compared to three to four-month-old rats. Lymphocytes and discrete brain areas showed a parallel pattern of age related alterations in the mitochondrial ETC complexes and CS. The analysis of such alterations is important in determining the ultimate basis of neuronal dysfunction associated with aging, and defining the nature of these changes may also help to develop therapeutic means to cure not only the elderly but also individuals suffering from certain organic or psychiatric disorders. PMID- 12652187 TI - Biological evidence for limits to the duration of life. AB - Projections of duration of life for humans based on mathematical models have led some researchers to claim that there is no lower limit to death rates or upper limit to life expectancy, and that a life expectancy of 100 will be achieved in the 21st century. To assess the biological plausibility of these claims, we examined temporal aspects of biological phenomena in three mammalian species. Our examination revealed that: (1) physiological declines associated with reproduction consistently occur at ages that are less than one-third of the median age at death, (2) physiological parameters associated with aging in humans lose eighty percent of their functional capacity by age 80, and (3) young versus old individuals can be distinguished by the pathologies detected at death. The biological evidence suggests that organisms operate under warranty periods that limit the duration of life of individuals and the life expectancy of populations. We use these findings to discuss the issue of limits to the duration of life and the validity of mathematical models used to forecast human longevity. PMID- 12652188 TI - Food restriction, pituitary hormones and ageing. AB - Reducing the intake of food in rodents inhibits body growth, retards most physiological ageing processes, delays the onset of pathology and prolongs life. Food restriction (FR) reduces pituitary hormone secretion and in consequence has been called 'functional hypophysectomy'. Direct life-long comparisons in the rat showed that hypophysectomy (HYP) (a complete absence of pituitary hormones) has a greater anti-ageing action than FR (a partial lack of pituitary hormones) on collagen, kidney and muscle. This suggests that pituitary hormones accelerate ageing. Recent American research on genetic variants of the mouse indicates that pituitary growth hormone (GH) may accelerate ageing and shorten life. Both the Snell and Ames dwarf mice have a deficiency of pituitary GH and live 50% longer than normal mice. The Snell dwarf mouse has retarded ageing of both collagen and immune functions. The Ames dwarf mouse has high antioxidant enzyme activities in liver and kidney. A transgenic human GH mouse is short lived, has a low activity of antioxidant enzymes in liver and kidney and an early development of disease in these organs. It is postulated that FR by reducing the secretion of pituitary hormones, such as GH, diminishes the oxidative damage of certain tissues, thereby delaying the development of age-related diseases in these tissues and by this means extends life. PMID- 12652190 TI - Diagnostic tissue elements in melanocytic skin tumors in automated image analysis. AB - In tissue counter analysis, digital images are divided into subregions (elements), and the digital information in each element is used for statistical analysis. In this study, we assessed the morphologic details of tissue elements that have turned out to be of diagnostic significance in the discrimination of benign common nevi and malignant melanoma. After creation of a data set based on a total of 12,000 cellular elements obtained from 100 benign common nevi and 100 malignant melanomas, classification and regression tree (CART) analysis was performed to differentiate between cellular elements of nevi and melanoma. In a second step, the slides were re-evaluated by the decision tree; cellular elements suggestive either for benign common nevi or for malignant melanoma were highlighted on zoomed images of the whole sections, and the individual elements were displayed in galleries. Eight groups of elements (so-called terminal nodes) seemed to indicate benign common nevi, whereas seven terminal nodes were suggestive for malignant melanoma. The elements of nodes suggestive for benign nevi largely contained nevus cells with amphiphilic cytoplasm intermingled with fibrillary material, whereas the elements of the nodes suggestive for malignant lesions often showed hyperchromatism, perinuclear halos, heavy pigmentation, or a lymphohistiocytic infiltrate. Tissue counter analysis automatically detects tissue elements that are in accordance with morphologic criteria used in conventional histopathology for diagnostic discrimination. PMID- 12652189 TI - S100A protein expression in the distinction between lentigo maligna and pigmented actinic keratosis. AB - Lentigo maligna (LM), a type of malignant melanoma in situ, and pigmented actinic keratosis (PAK) may have similar clinical appearances but are different in prognosis and treatment. Diagnosis is established by skin biopsy. In certain cases, microscopic features may be very similar in both entities, making it difficult to determine whether the pigmented atypical cells are keratinocytes or melanocytes. Immunohistochemical markers can be useful for the identification of melanocytes in these cases. There are limitations to the use of some standard immunohistochemistry markers, however. S100 proteins are a varied group of proteins that are of special interest because of their dysregulated expression in neoplastic disorders. Their expression is changed during malignant transformation, progression, and/or metastasis in various cell lines and tumors, including melanomas. Our study analyzed the expression of several of the S100 protein subtypes (S100A2, S100A6, and S100A8/A9 or A12) in 38 LM cases and 44 PAK cases to define their potential value in the distinction between these entities together with their role in the development of early malignant melanoma of the skin. The results showed an upregulation of S100A2 protein in atypical keratinocytes in PAK and in normal keratinocytes adjacent to melanoma cells in LM. There was also an upregulation of S100A8/A9 or A12 protein, as detected by the antibody MAC387, in normal keratinocytes adjacent to both atypical keratinocytes and melanocytes in PAK and LM, respectively. There were statistically significant differences in the level of positive cells and in the pattern of immunoreactivity for anti-S100A2 and MAC387 in each entity, however. Moreover, the findings of our study support the notion that melanocyte keratinocyte interactions are abnormal in both of these disease entities and may be involved in their progression. PMID- 12652191 TI - Dendritic cells and pattern of cytokines in paracoccidioidomycosis skin lesions. AB - We demonstrated and quantified by immunohistochemistry epidermal Langerhans cells, CD34+ dermal dendrocytes (DDs), and cells expressing TNFalpha, interferon gamma (IFNgamma), IL-5, and IL-10 in skin lesions of paracoccidioidomycosis (PCM). Sixty-one biopsies were classified in three groups according to the pattern of tissue response: Group 1, well-organized granuloma; Group 2, poorly organized granuloma; and Group 3, both kinds of granuloma. Langerhans cells had short and irregular dendrites in all groups and were decreased in number in Groups 1 and 2. CD34+ DDs did not differ in number from the control group. Group 1 was characterized by many cells expressing IFNgamma. Groups 2 and 3 exhibited large numbers of cells expressing IL-5 and IL-10. The data obtained suggest that well-organized granulomas reflect a better cellular immune response, and the large number of cells expressing IL-5 and IL-10 in Group 2 indicate an ineffective response in PCM skin lesions. Both kinds of granuloma in the same cutaneous lesion probably represent an intermediate response between the anergic and hyperergic poles. Group 3 also showed higher numbers of cells expressing TNFalpha when compared with the control group. Some cells expressing TNFalpha were dendritic and localized around the granuloma similar to the factor XIIIa+ DD localization that we previously described. PMID- 12652192 TI - Granuloma annulare and malignant neoplasms. AB - The relationship between granuloma annulare (GA) and malignant neoplasms is uncertain. Several case reports and clinical correlation studies have addressed this issue; however, no review articles have summarized those reported cases and statistically analyzed the association. Fourteen case reports and two correlation studies directly addressing the relationship of GA and malignant neoplasms were found from the Medline database from 1966-2001. A total of 16 cases from case reports were available with the mean age of 54. Most of the GA patients had an atypical clinical presentation. The time between the onset of GA and the discovery of the malignant neoplasms varied from 18 months before (median = 5 months) to 7 years after (median = 42 months). More than half of those cases were associated with lymphoma. No definite relationship was found between GA and malignant neoplasms. Older patients with skin lesions not clinically typical of GA but that histologically resemble classic GA, may be considered for investigation of underlying malignant lymphoma or other malignancies. PMID- 12652193 TI - Prurigo pigmentosa: a distinctive inflammatory disease of the skin. AB - More than 200 patients with prurigo pigmentosa, a disease described first by Nagashima in 1971, have been reported on in Japan, but only 28 non-Japanese patients have come to notice as of today. In order to establish reliable, repeatable criteria for diagnosis of the disease, we studied 25 patients with prurigo pigmentosa and reviewed the literature pertaining to it as recorded in another 182 patients.Clinically, prurigo pigmentosa presents itself as pruritic urticarial papules, papulovesicles, and vesicles arranged in reticular pattern and distributed symmetrically on the back, neck, and chest. Lesions involute in a matter of days, leaving behind netlike pigmentation. Exacerbations and recurrences are the rule. Histopathologically, prurigo pigmentosa begins with a superficial perivascular infiltrate of neutrophils. Shortly thereafter, neutrophils are scattered in dermal papillae and then sweep rapidly through an epidermis in which spongiosis, ballooning, and necrotic keratocytes are accompaniments. En route, abscesses may form in the surface epithelium. Very soon, eosinophils and lymphocytes come to predominate over neutrophils in a dermal infiltrate that assumes a patchy lichenoid pattern. Intraepidermal vesiculation follows on spongiosis and ballooning and, sometimes, subepidermal vesiculation on vacuolar alteration at the dermo-epidermal junction. As the epidermis becomes hyperplastic, parakeratotic, and slightly hyperpigmented, melanophages begin to appear in the dermis. Studies by immunofluorescence are negative invariably. Dapsone or minocyclin are effective treatments; both of those agents inhibit migration and/or function of neutrophils. The cause and pathogenesis have yet to be determined. Prurigo pigmentosa is unique among inflammatory diseases of the skin and the singularity of it is manifest both clinically and histopathologically. PMID- 12652194 TI - Familial basaloid follicular hamartoma: lesional characterization and review of the literature. AB - Basaloid follicular hamartoma (BFH) is a rare cutaneous lesion associated with the acquisition of small papules that remain stable for many years. Basaloid follicular hamartoma lesions can present sporadically or as part of an inherited syndrome. Occasionally, biopsies of BFH lesions are interpreted as basal cell carcinoma (BCC), which necessitates complete removal of the lesion. In this report, we characterize a case of a familial BFH syndrome and discuss the clinical, histologic, and molecular features of BFH lesions that help to distinguish it from BCC. The BFH lesions in our patients remained stable for many years. Histologically, BFH lesions exhibit fewer mitoses and decreased single cell necrosis when compared with BCC. Immunohistochemical staining for the proliferation markers proliferating cell nuclear antigen and Ki-67 demonstrated less staining in BFH than in BCC. In addition, levels of PTCH (patched) mRNA were increased relative to unremarkable epidermis in familial BFH lesions but to a lesser degree and in a different pattern than that seen in BCC. In summary, familial BFH can be distinguished from BCC based on clinical, histologic, and molecular features and is associated with deregulation of the PTCH pathway. Basaloid follicular hamartoma may represent an indolent lesion within the spectrum of basaloid epithelial neoplasms associated with deregulation of the PTCH signaling pathway. We discuss this case in parallel with a growing body of literature that supports the nosologic designation of BFH. PMID- 12652195 TI - Lipomatous apocrine mixed tumor of the skin. AB - The common mesenchymal elements of apocrine mixed tumor of the skin are mucin and chondroid tissue. An apocrine mixed tumor of the skin with extensive adipose content is reported. A 56-year-old Japanese man presented with a subcutaneous mass in the glabella region that was 1 cm in diameter and had developed within the preceding 5 years. Light microscopic examination showed a well-circumscribed and encapsulated subcutaneous tumor composed of tubular structures with apocrine secretion and mucin as well as extensive adipose tissue. This case, in light of another recently reported case, indicates that adipose tissue can be the major mesenchymal element in apocrine mixed tumor of the skin. PMID- 12652196 TI - Neutrophil-rich anaplastic large cell lymphoma presenting in the skin. AB - A neutrophil-rich anaplastic large cell lymphoma (ALCL) presented in the skin of a 47-year-old Japanese woman. The solitary cutaneous lesion was an eroded, 10-mm, dome-shaped nodule involving the skin of her left upper arm. Histologically, it showed a proliferation of pleomorphic, anaplastic, large tumor cells with nuclei of various shapes, including embryo-shaped, Reed-Sternberg cell-like binucleated, and wreath-shaped multiple nuclei, in the dermis and subcutaneous tissues. There was an admixture of neutrophils ranging from about 30% to more than 50% per field. Immunophenotypically, the neoplastic cells were positive for CD30, CD4, leukocyte common antigen, anaplastic lymphoma kinase-1, epithelial membrane antigen, and granzyme B. She had no peripheral neutrophilia. The lesson was excised, and the site on the left upper arm was irradiated. Six and a half months after diagnosis, however, swelling of a left axillary lymph node appeared; it also showed a proliferation of anaplastic large tumor cells admixed with numerous neutrophils ranging from about 25% to more than 60% per field. Southern blot analysis of T-cell receptor gene rearrangement revealed a clonal band. The patient was treated with six courses of cyclophosphamide, doxorubicin, vincristine, and prednisolone chemotherapy with complete remission. Seventeen cases of neutrophil-rich ALCL arising in the skin, lymph node, muscle, testis, and skull bone were reviewed. This form may possibly be induced by trauma or irritation of conventional ALCL, although the true mechanism for the infiltration of neutrophils is still unclear. PMID- 12652197 TI - Mycobacterium marinum infection simulating interstitial granuloma annulare: a report of two cases. AB - We report two cases of Mycobacterium marinum infection that histologically simulated interstitial granuloma annulare (GA). In one case, an infectious etiology was not suspected in histologic sections, but a tissue culture performed during the patient's clinic visit identified M. marinum, and a subsequent Fite stain revealed mycobacteria. Interstitial granulomatous dermatitis is a rare presentation for cutaneous nontuberculous mycobacteria and has yet to be attributed specifically to M. marinum. In both immunocompetent and immunosuppressed patients, infection with M. marinum should be considered in lesions histologically resembling interstitial GA, particularly when there is clinical suspicion for an infectious process. PMID- 12652198 TI - Sweet's syndrome-like blastomycosis. AB - Cutaneous North American blastomycosis is characterized clinically by verrucous nodules and histologically by pseudoepitheliomatous hyperplasia, intraepidermal neutrophilic microabscesses, and a dermal mixed inflammatory cell infiltrate containing giant cells. We describe a patient who presented clinically with erythematous nodules and plaques on the lower extremities characterized histologically by a diffuse neutrophilic infiltrate, with lack of epidermal hyperplasia. The lesions were clinically and histologically reminiscent of Sweets syndrome. On close microscopic inspection scattered histiocytes and multinucleated giant cells were present in the dermis, and fungal stains demonstrated budding yeast forms consistent with Blastomyces sp. PMID- 12652199 TI - The man behind the eponym: Lucien Marie Pautrier--Pautrier's microabscess. AB - Lucien-Marie Pautrier, recognized during his life as a world-renowned dermatologist and venereologist, also wrote prolifically, established and headed a leading department at the medical school of Strasbourg, built an imposing structure to house the department, traveled widely in both Europe and the United States and founded the Societe des Amis de la Musique at Strasbourg. He did all this while and between the two tragic World Wars that engulfed his beloved France. In the United States, his name is eponymically associated with Pautrier's microabscesses in mycosis fungoides despite the fact that he did not first describe them. PMID- 12652200 TI - Lymphoid lost and found. AB - We present the clinicopathologic features of a case initially interpreted as lymphocytoma cutis that was later determined to be lymphoepithelioma-like carcinoma of the skin. The histologic presence of nodular aggregates of undifferentiated epithelioid cells surrounded by a dense, reactive lymphocytic infiltrate should prompt consideration of the diagnosis of lymphoepithelioma-like carcinoma of the skin. This lesion should be distinguished from lymphocytoma cutis, B-cell lymphoma, and cutaneous lymphadenoma. PMID- 12652201 TI - Ducking stray "magic bullets": a Melan-A alert. PMID- 12652202 TI - Is melanocytic matricoma a bona fide entity or is it just one type of matricoma? PMID- 12652203 TI - Cellular storiform collagenoma. PMID- 12652204 TI - Perineurioma with adipocytes (lipomatous perineurioma). PMID- 12652205 TI - Response to "Intratumoral fat in neurofibroma". PMID- 12652206 TI - Median raphe cyst with ciliated cells: report of a case. PMID- 12652207 TI - Ectopic hidradenoma papilliferum: a case report and literature review. PMID- 12652208 TI - Clinical pathologic challenge. Confluent and reticulate papillomatosis of Gougerot and Carteaud. PMID- 12652209 TI - Postoperative arrhythmia. AB - Postoperative arrhythmia is a major cause of morbidity and mortality after cardiac surgery for congenital heart disease. Rhythm disturbances that may be well tolerated in a normal heart often cause hemodynamic instability when they occur in the immediate postoperative period. In the face of pre-existing myocardial dysfunction resulting from preoperative pressure or volume overload, patients with congenital heart conditions are especially vulnerable to rhythm disturbances after cardiac surgery. Cardiopulmonary bypass, intraoperative injury to the conduction system and myocardium, postoperative metabolic abnormalities, electrolyte disturbances, and increased adrenergic tone in response to the stress of the surgery or inotropic agents are also known factors associated with increased risk of arrhythmia in the immediate postoperative period. Surgically related arrhythmia can also present in the late postoperative period, particularly in association with surgical incision sites and surgically induced hemodynamic abnormalities. Early and late postoperative arrhythmias are important risk factors for morbidity and mortality after surgical treatment of many forms of congenital heart disease. This review describes the incidence of the most common forms of arrhythmia and recent advances in their diagnosis and treatment. PMID- 12652210 TI - Future of interventional cardiology in pediatrics. AB - A trend toward use of less invasive, nonsurgical approaches to the treatment of congenital heart disease has developed. Although transcatheter valve replacements and ventricular septal defect closures have already been performed in children, many other techniques and devices are being developed for the palliation of children with a single ventricle. Several groups have already established catheter-based procedures for the creation of Fontan communications, and others have performed nonsurgical pulmonary artery banding. Additionally, fetal catheter based interventions are being developed for the treatment of severe congenital heart disease in utero. Because this trend toward catheter-mediated treatment is certain to continue, care must be taken to regulate safely the introduction of novel techniques and devices into clinical use in pediatric cardiology. PMID- 12652211 TI - Cardiomyocyte injury to transplant: pediatric management. AB - Cardiomyocyte injury in pediatric patients has a vast number of causes, which are often distinct from the causes of adult heart failure. However, the management of pediatric heart failure and heart transplantation has generally been inferred from adult studies. New therapies show great promise for the neurohormonal regulation of heart failure and the ability to control immunosuppression after heart transplantation. Large, randomized, multicenter, controlled clinical trials are needed to determine the efficacy of these therapies in this population. This article reviews the current recommendations and evidence-based medicine, where available, for the medical management of myopathic dysfunction and transplantation in pediatric patients. PMID- 12652212 TI - Resources, use, and regionalization of pediatric cardiac services. AB - Care for children with congenital heart disease requires specialized services and various healthcare resources. The purpose of this article is to provide an updated overview of healthcare resources for infants and children with heart disease. In 2001, there were 1609 certified pediatric cardiologists in the United States. The ratio was approximately 45,000 children younger than 18 years per pediatric cardiologist. It is estimated that more than 19,000 cardiac surgeries are performed in children younger than 18 years in the United States each year. This article also reviews the effect of patient characteristics on access to healthcare and use of pediatric cardiac services, and discusses issues related to optimal use of these resources and the development of an organized approach toward service management by regionalization. The authors believe that improved access to high-quality facilities and providers coupled with thoughtful changes in the healthcare delivery system represent an excellent opportunity for optimizing outcomes for children with heart disease. PMID- 12652213 TI - Mitral valve replacement versus repair in 2003: where do we stand? AB - Mitral valve repair as a proportion of all mitral valve procedures continues to become more prominent in this country. As the procedures are performed more frequently and concomitantly surgeons become more comfortable performing these repairs, risks and benefits (compared with mitral valve replacement) have become better defined. The patient population and valve morphology that are most favorable for the repair approach have also been better defined, and new techniques (including minimally invasive approaches) have been reported. A prominent question remains: what to do about ischemic mitral regurgitation. PMID- 12652214 TI - Diagnosis and management of bacterial endocarditis in 2003. AB - The diagnosis of infective endocarditis has been notoriously difficult. Over the last decade, the modified Duke criteria have assumed an increasingly important role in the early detection of this often occult disease. Echocardiography has assumed increasing importance. Transesophageal echocardiography is recognized as more sensitive and specific than transthoracic echocardiography at detecting vegetations less than 10 mm in diameter. Vegetations greater than 10 mm in diameter are thought to be at increased risk of embolizing. Combined medical and surgical medical management result in the lowest mortality for those patients with hemodynamic compromise. PMID- 12652215 TI - Tricuspid valvular disease in the patient with chronic pulmonary thromboembolic disease. AB - Chronic thromboembolic pulmonary hypertension is associated with right ventricular dilatation, high right-sided filling pressures, and functional tricuspid regurgitation. The tricuspid regurgitation resulting from this disease has been postulated to be caused by tricuspid annular dilatation with displacement of the papillary muscles. Pulmonary endarterectomy is an operation that corrects the pulmonary hypertension resulting from chronic thromboemboli. As a result of this operation, most patients show significant improvement in tricuspid valve function. Thus, pulmonary endarterectomy is one of the few cardiac operations in which surgery remote to a valve restores valve function. This review analyzes the factors responsible for changes in tricuspid valve regurgitation after pulmonary endarterectomy and provides a strategy to predict the small subset of patients who will not show improvement in tricuspid valve function after this operation. PMID- 12652216 TI - Current thinking in stentless valve surgery. AB - Aortic valve replacement for aortic stenosis represents a tremendous achievement in the management of cardiac disease. However, despite 4 decades of use, the ideal substitute for the diseased aortic valve is still not agreed upon. Stentless aortic valves represent the optimum in hemodynamic performance. This article reviews the current thinking in stentless aortic valve surgery. PMID- 12652217 TI - Valvular surgery in the transplanted heart. PMID- 12652218 TI - Cardiac transplantation 2003. PMID- 12652219 TI - Quality of life after heart transplantation: are things really better? AB - Studies of quality of life (QOL) in heart transplant recipients have been published during the last 2 decades. More recent studies of QOL outcomes have built on previous research. Relationships between posttransplant complications and QOL, longitudinal studies of intermediate and long-term QOL, QOL in patients awaiting transplant (bridged to transplant with a left ventricular assist device), and intervention studies to improve QOL have been published recently. Is QOL better from before to after heart transplantation? The current body of literature suggests that QOL is better overall. However, change in QOL (both positive and negative) is variable based on demographic characteristics, clinical problems, QOL domain, time posttransplant, and other life events. Moreover, although more recent reports have begun to examine unanswered questions, much work remains to be done. Future studies need to be scientifically rigorous, using definitions of QOL with identification of domains to be studied; prospective, multisite, longitudinal study designs; large sample sizes; reliable and valid instruments; and appropriate statistical techniques. As changes in QOL outcomes and risks for poor QOL outcomes are identified, more intervention studies need to be developed to assist patients toward better QOL. PMID- 12652220 TI - Evaluation of pulmonary hypertension in heart transplant candidates. AB - Assessment of the pulmonary circulation and right ventricular function is a cornerstone in the evaluation of the patient as a potential heart transplant recipient. The importance of pulmonary hypertension is linked to outcomes in the posttransplant period. Preoperative pulmonary vascular resistance is an independent risk factor for early death after heart transplantation. Pulmonary hypertension can be classified as reversible, or irreversible if not rapidly responsive to pharmacologic maneuvers. However, in most patients, the major component is likely to reverse with vasodilators, because of the central role played by the endothelium in the control of pulmonary vascular tone. To discriminate between patients with reversible and irreversible pulmonary hypertension, provocative therapies are used, and baseline and the postprovocation hemodynamic parameters are measured. To date, there is no reliable hemodynamic threshold beyond which right ventricular failure is certain to occur, nor are there values below which right ventricular failure is always avoidable. Because of this uncertainty, it becomes clear that only through careful preoperative assessment can this life-threatening condition be recognized preoperatively and, hence, managed in the posttransplant recovery period. PMID- 12652221 TI - Patient selection for assist devices: bridge to transplant. AB - Patients in severely progressed states of heart failure can be bridged to successful heart transplantation with mechanical assist devices. Experience has demonstrated that patient selection and timing of device implantation are crucial for obtaining acceptable results when using this expensive technology. The degree of irreversible secondary organ dysfunction before re-establishing adequate cardiac output determines the chance of reaching transplantation. Patients who recover during support from all sequelae of end stage heart failure have an excellent outcome after heart transplantation. PMID- 12652222 TI - Mechanical circulatory support device database of the International Society for Heart and Lung Transplantation. AB - Over the last 2 decades, mechanical circulatory support devices have been developed at a rapid pace with the goal of supporting patients with advanced heart failure as a bridge to cardiac transplantation, a bridge to recovery, and an alternative to transplantation, also called chronic or destination therapy. The current generation of devices provides a differentiated spectrum of circulatory support. The major limitations are infection, coagulopathies, and device dysfunction. The Scientific Council on Mechanical Circulatory Support of the International Society for Heart and Lung Transplantation has established an international database (http://www.ishlt.org/regist_mcsd_main.htm) to generate critical data to advance knowledge about effectiveness of mechanical circulatory support device therapy for one of the most difficult and costly contemporary medical problems, the malignant syndrome of advanced heart failure. PMID- 12652223 TI - Allosensitization in heart transplantation: implications and management strategies. AB - The detection of anti-human leukocyte antigen (HLA) donor-specific antibodies has been associated with a variety of clinical syndromes that determine short-term and long-term outcomes of cardiac transplant recipients, including an increased incidence of early and more severe allograft rejection and cardiac allograft vasculopathy. Recent surveys indicate marked heterogeneity in clinical protocols for detection and management of sensitization in heart transplantation. The commonly performed complement-dependent cytotoxicity assay is insensitive compared with newer methods such as flow cytometry for antibody screening. The imperative exists to create strategies that can decrease the level of sensitization and increase the likelihood for a negative crossmatch. In this effort, several strategies have been suggested, including administration of intravenous immunoglobulin, apheresis, and combination therapies using potent immunosuppression, particularly with cyclophosphamide. The future of managing allosensitization may be in induction of a chimeric state to allow graft tolerance. PMID- 12652224 TI - Cystoid macular degeneration in chronic central serous chorioretinopathy. AB - PURPOSE: To describe the optical coherence tomography (OCT) and fluorescein angiography findings in the macula of eyes with chronic central serous chorioretinopathy (CSC) and reduced central vision. METHODS: Using OCT, clinical examination, and fluorescein and indocyanine green (ICG) angiography, the authors examined eight eyes of seven patients with CSC, an attached macula, and reduced central vision of 20/200 or worse. All had a history of chronic CSC with resolution of the subretinal fluid in the macular area and poor vision. RESULTS: Patient ages ranged from 55 to 82 years (mean, 66 years). All eight eyes had some parafoveal, patchy RPE atrophy with corresponding transmission hyperfluorescence (window defect) on fluorescein angiography. Five eyes also had a window defect in the foveal area. With OCT, the foveal area revealed variable areas of cystoid change and atrophy in seven of the eight eyes. In four of these eyes, the cystoid changes were not seen on clinical examination or fluorescein angiography. The seven eyes with cystoid changes imaged with OCT had no intraretinal leakage of fluorescein in the foveal region. The authors categorized these eyes as having cystoid macular degeneration (CMD). One other eye had foveal thinning or atrophy without cystoid changes. CONCLUSIONS: Intraretinal cystoid spaces without intraretinal leakage, or CMD, was a common finding in eyes with chronic CSC and reduced central vision after resolution of subretinal fluid. OCT was useful to establish the presence of CMD and foveal atrophy, even when these changes were not clearly evident on clinical examination or fluorescein angiography. Chronic foveal detachment and antecedent intraretinal leakage were proposed to be the mechanisms for the development of the changes. CMD in conjunction with foveal atrophy was an important clinical finding to account for the poor visual outcome in patients with CSC. PMID- 12652225 TI - Intraoperative ketorolac and eye pain after viteoretinal surgery: a prospective, randomized, placebo-controlled study. AB - PURPOSE: To compare the efficacy of one intraoperative dose of intravenous ketorolac tromethamine to saline placebo in controlling postoperative eye pain, nausea, and sedation following vitreoretinal surgery. STUDY DESIGN: Prospective, randomized, placebo-controlled clinical trial. METHODS: One hundred fourteen vitreoretinal surgical patients were randomized over a 7-month period to a single intraoperative intravenous dose of ketorolac tromethamine or placebo. All patients underwent surgery with intravenous sedation and retrobulbar anesthetic injection and had access to a standard postoperative analgesic regimen. After 2 and 5 hours, eye pain, nausea, and sedation levels were recorded using a standard visual analog scale. Analgesic requirements were tabulated. RESULTS: Patients given ketorolac were significantly less likely to have postoperative eye pain than were placebo-treated patients (P = 0.0043). Fewer ketorolac patients required postoperative analgesia than placebo patients did (P < 0.0001). Four ketorolac patients (7%) required a narcotic compared to 11 placebo patients (20%) (P < 0.0001). Fewer ketorolac patients experienced nausea and sedation than placebo patients did, but this difference was not statistically significant. CONCLUSION: Ketorolac tromethamine is an effective nonnarcotic choice for postoperative pain control following vitreoretinal surgery. Pain control is particularly important in this group of patients, whose compliance with postoperative procedures and positioning may significantly affect surgical outcome. PMID- 12652227 TI - Results of retinal detachment surgery in Marfan syndrome in asians. AB - PURPOSE: To report the anatomic and visual results of retinal detachment in patients with Marfan syndrome in an Asian population. METHOD: A retrospective review of all patients with Marfan syndrome and retinal detachment operated on by a single surgeon was conducted. RESULTS: Thirteen eyes of 12 patients were reviewed. Eleven patients were males, with an average age of 24.6 years. Three eyes had a history of trauma. Ten eyes were successfully treated with scleral buckle and encircling band. Three eyes were also treated with vitrectomy, scleral buckle, and encircling band. One hundred percent success was achieved with only one surgery. Best-corrected visual acuity of at least 20/40 was achieved in 7 of 12 eyes (58.3%). Eight of 12 eyes (66.7%) had at least two lines of improvement on the Snellen chart after 6 months. CONCLUSIONS: With appropriate surgical intervention, an excellent anatomic reattachment rate and good visual outcome can be achieved in Marfan syndrome patients with retinal detachment regardless of their lens status. PMID- 12652226 TI - Anecortave acetate as monotherapy for the treatment of subfoveal lesions in patients with exudative age-related macular degeneration (AMD): interim (month 6) analysis of clinical safety and efficacy. AB - PURPOSE: To evaluate clinical safety and efficacy of the angiostatic agent anecortave acetate for treatment of subfoveal choroidal neovascularization secondary to AMD. METHODS: 128 patients were randomized to placebo treatment or one of three anecortave acetate doses. Study medication was administered as a posterior juxtascleral injection onto the posterior scleral surface. Best corrected logMAR vision was obtained at baseline and follow-up visits. Fluorescein angiograms were evaluated for eligibility before enrollment and posttreatment. RESULTS: Six months after a single treatment, visual acuity (mean change from baseline logMAR values) was significantly better (P = 0.003) after anecortave acetate 15 mg than placebo. More patients treated with anecortave acetate 15 mg than placebo maintained vision (88% versus 70%, P = 0.080), especially those with predominantly classic lesions (92% versus 65%, P = 0.021). Anecortave acetate 15 mg inhibited lesion growth significantly better than placebo (P = 0.001). Trends favoring the other doses over placebo were observed for vision preservation and lesion inhibition, but statistical significance was not achieved. The Independent Safety Committee overseeing this study identified no clinically relevant treatment-related changes. CONCLUSION: Anecortave acetate 15 mg is safe and effective for preserving or improving vision and for inhibiting lesion growth in patients with subfoveal AMD. PMID- 12652229 TI - Rhegmatogenous retinal detachment in the fellow phakic eyes of patients with pseudophakic rhegmatogenous retinal detachment. AB - PURPOSE: To determine the risk and clinical characteristics of rhegmatogenous retinal detachment (RD) in the fellow phakic eyes of patients with a pseudophakic retinal detachment (PPRD) in one eye. METHODS: The authors conducted a computer generated chart review to find 64 consecutive patients with a PPRD in one eye and were phakic in the fellow eye. The clinical information was collected on all patients to investigate the risk and clinical characteristics of RD in the fellow eyes while still phakic. Information was also obtained on the fellow eyes that underwent cataract surgery and developed a PPRD. RESULTS: Sixty-four patients with a PPRD in one eye were phakic in the fellow eye. During an average follow-up of 57.4 months, five (7.8%) fellow eyes developed retinal detachment while still phakic. In addition to the five eyes with a phakic RD, 10 originally phakic fellow eyes underwent cataract surgery. Of these, one (10%) suffered an RD. CONCLUSION: Fellow eyes of patients with a PPRD have a significant risk of RD even if they do not undergo cataract surgery. PMID- 12652228 TI - Endoscopy for evaluation and treatment of the ciliary body in hypotony. AB - PURPOSE: To determine the clinical utility of the intraocular videoendoscope for evaluation of the ciliary body in eyes with hypotony and as a visualization tool for surgery of the ciliary body. METHOD: A gradient index of refraction rod endoscope was used to evaluate the ciliary body in 14 patients. Videotapes and charts were reviewed retrospectively to correlate the appearance of the ciliary body and to analyze the clinical findings and surgical results. Videoendoscopic surgery to remove fibrous tissue from the ciliary processes was performed in nine eyes. RESULTS: The endoscope produced a clear detailed image of the ciliary body in a state that is undisturbed by scleral depression or other forms of mechanical distortion. In eyes without hypotony the normal ciliary processes are uniformly pigmented with a velvet-like surface at high magnification. White surface changes in the peak and crest of individual ciliary processes (white-caps) were seen in 11 of 14 patients with hypotony. At high magnification the white surface changes had a brush-like texture integral to the ciliary processes. Other findings in hypotony were fibrous covering of the ciliary body in 10 patients, loss of pigment epithelium in 5, traction elongation of the ciliary processes in 8, vascular loops in 4, pigmented sheets in 3, and areas of missing ciliary processes in 1. White caps were present in seven of the nine patients who had surgery for fibrous traction of the ciliary processes and hypotony and three patients with hypotony not due to other endoscopically visible abnormalities of the ciliary processes. Endoscopically visualized dissection of ciliary body in 9 patients resulted in normal or elevated tension immediately postoperatively in 7 (78%). At the most recent visit only three of the patients had normal tension, but for these nine patients the pressures were marginally, significantly (P = 0.08) higher compared to the preoperative pressures. CONCLUSION: The evaluation and management of hypotony is enhanced by the use of intraocular videoendoscopy. Clinical findings of whitecaps and traction elongation of the ciliary processes were associated with ocular hypotony. The endoscope facilitated surgery for dissection and removal of fibrous tissue over the ciliary processes. Further studies are needed to better understand the pathophysiology of ocular hypotony. PMID- 12652230 TI - Penetrating keratoplasty surgery combined with vitrectomy after failing previous corneal surgery. AB - PURPOSE: The preoperative and postoperative factors influencing visual outcome were analyzed in 15 eyes of 15 patients with graft opacity after keratoplasty associated with vitreoretinal disease who underwent combined surgery of fresh corneal retransplantation and vitrectomy. MATERIALS AND METHODS: The data obtained consisted of diagnosis, preoperative visual acuity, corneal and vitreoretinal findings at the time of surgery, interval between the first visit and surgery, intraocular pressure before surgery, gonioscopic findings, results of bacterial culture of surgical specimens, surgical procedure for vitrectomy, corneal and vitreous findings after surgery, visual acuity 6 months after surgery, intraocular pressure after surgery, and additional surgical techniques. RESULTS: The cause of corneal opacity was graft rejection in all patients, who had been treated with high-dose administrations of steroids and cyclosporin. The preoperative diagnosis was proliferative vitreoretinopathy (PVR) in eight eyes and fungal endophthalmitis in seven eyes. Corneal transparency was achieved in 7 eyes (46.6%) 6 months after surgery. Vitreoretinal findings improved in 9 eyes (60%) and PVR recurred in 6 eyes (40%). Visual acuity improved in 7 eyes (46.6%), did not change in 2 eyes (13.3%), and deteriorated in 6 eyes (40%). The six eyes with decreased visual acuity developed phthisis bulbi. Preoperative intraocular pressure was 2.1 mmHg on average in the phthisis bulbi group, significantly lower than in the group with superior prognosis. Goniosynechia was noted before surgery and did not improve after surgery in all six eyes. CONCLUSIONS: The outcome was poor in eyes with goniosynechia and ocular hypotony, and combined surgery is not indicated for either anatomic or visual preservation in such cases. Care should be taken not to overlook intraocular infection in patients undergoing immunosuppressive therapy against graft rejection. The early detection of retinal detachment is also important in eyes exhibiting hypotony after surgery. PMID- 12652231 TI - Outcome of combined penetrating keratoplasty with vitreoretinal surgery for management of severe ocular injuries. AB - PURPOSE: To assess the effectiveness of a combined procedure (pars plana vitrectomy with temporary keratoprosthesis, vitreoretinal surgery, and penetrating keratoplasty) and to determine which factors influence corneal transplant success rates. METHODS: Records for 34 severely injured eyes of 31 consecutive patients who underwent penetrating keratoplasty in combination with vitreoretinal surgery between 1991 and 1998, with a follow-up of at least 12 months, were evaluated retrospectively. Analysis was focused on ocular history, functional and anatomic anterior and posterior segment outcome, and complications. RESULTS: Penetrating keratoplasty was performed in 10 eyes (29%) within 8 weeks of trauma and in 24 eyes (71%) as a secondary procedure. Initial improvement in visual acuity (VA) was achieved in 47% of eyes; at the end of the follow-up, 74% of the eyes had attained equal or better VA than the initial postsurgical VA. Reasons for poor final VA were hypotony or phthisis (35%) and recurrent retinal detachment (11%). Corneal grafts remained clear for 1 year in 65% of eyes. Transplant failure was less in eyes that did not need a permanent silicone oil tamponade, when the retina was attached before surgery, and in eyes that were grafted later after trauma and received no further surgery. These were some conditions that were associated with a longer mean graft survival time, but without statistical significance. There was no difference in eyes according to the trauma mechanism, preoperative intraocular pressure, or graft size. CONCLUSIONS: Penetrating keratoplasty in severely injured eyes is often complicated by ciliary body malfunction and secondary transplant failure. Although the functional outcome of a combined procedure is limited by primary and secondary tissue destruction, preserving ambulatory vision is possible and thus improves the quality of life, at least in patients with single remaining eyes. PMID- 12652232 TI - Anatomic and functional results of vitrectomy and long-term intraocular tamponade for stage 2 macular holes. AB - PURPOSE: To investigate the results of pars plana vitrectomy combined with long term intraocular tamponade in patients with a stage 2 macular hole. METHODS: In a retrospective study, 50 consecutive eyes operated on for stage 2 macular hole were reviewed. The surgical technique included pars plana vitrectomy, separation of the posterior hyaloid, and intraocular tamponade with either 12.5% perfluropropane (C(3)F(8)) gas or silicone oil. Patients treated with C(3)F(8) were instructed to keep face-down positioning for 3 or 4 weeks. Removal of silicone oil took place 4 or 5 weeks after the vitrectomy. RESULTS: Closure of the macular hole was achieved in all 50 eyes after one operation. The mean postoperative best-corrected distance visual acuity was 20/32 (range, 20/63 20/20), with 49 eyes (98%) having a postoperative visual acuity of 20/50 or better. Visual acuity improved in all eyes postoperatively, with a mean gain of 4.84 +/- 1.95 ETDRS lines. CONCLUSION: For stage 2 macular holes, vitrectomy combined with long-term intraocular tamponade can result in a very favorable anatomic and functional outcome that is as good as or better than the results described for other modalities. This approach simplifies the surgical technique, and both adjuvant-related complications and potential retinal damage related to internal limiting membrane peeling or use of indocyanine green are avoided. PMID- 12652233 TI - Acute posterior multifocal placoid pigment epitheliopathy associated with a systemic necrotizing vasculitis. AB - PURPOSE: The authors present a case of acute posterior multifocal placoid pigment epitheliopathy (APMPPE) associated with a systemic necrotizing vasculitis with mixed features of Wegener granulomatosis and polyarteritis nodosa (PAN). METHODS: Case report. RESULTS: A 29-year-old woman developed a severe nosebleed, followed by a low-grade fever, night sweats, and a productive cough. One month later, she began experiencing high fevers, headache, myalgia, neck stiffness, and abdominal pain as well as bilateral blurred vision from APMPPE. Systemic evaluation revealed nasal ulcerations, bilateral pleural effusions, and a bilateral maxillary and ethmoid sinusitis, consistent with Wegener granulomatosis. However, ANCA testing was negative, and a renal and mesenteric angiogram showed aneurysmal dilatations suggestive of PAN. Her ocular and systemic symptoms markedly improved with systemic corticosteroids. CONCLUSIONS: The cause of APMPPE is unknown. This case of APMPPE associated with systemic necrotizing vasculitis provides support for the choroid as being primarily involved by a diffuse vasculitic process that interrupts choroidal perfusion and causes the characteristic fundus findings in APMPPE. PMID- 12652234 TI - Optical coherence tomography findings in the macula after treatment of rhegmatogenous retinal detachments with spared macula preoperatively. AB - PURPOSE: To present the postoperative optical coherence tomography (OCT) findings of the macula in rhegmatogenous retinal detachment with spared macula preoperatively. METHODS: Eleven of 46 patients who underwent surgery for rhegmatogenous retinal detachment had an uninvolved macula preoperatively. Scleral buckling without intravitreous gas injection was the operation used in all eyes. All 11 patients were examined before and after treatment with slit-lamp biomicroscopy, indirect ophthalmoscopy, fundus photography, fluorescein angiography, and OCT. RESULTS: In 3 of the 11 patients with rhegmatogenous retinal detachment and uninvolved macula preoperatively, fluid was identified with OCT in the macula after successful treatment. OCT showed that the fluid gradually diminished and finally disappeared 5 to 7 months after the scleral buckling procedure. The fluid in the macula became visible with slit-lamp biomicroscopy in only one of the three patients. CONCLUSION: Subretinal fluid in a preoperatively uninvolved macula can be found after successful treatment of rhegmatogenous retinal detachment. If the fluid is located in the fovea, the visual acuity decreases and does not reach the preoperative levels. OCT identifies the presence of fluid and contributes to the study of the fluid's evolution. PMID- 12652235 TI - Force comparison of air currents produced by a standard and modified infusion cannula. AB - OBJECTIVE: To develop and characterize an infusion cannula that reduces the intraocular jet forces created during fluid-air exchange. METHODS: A new infusion cannula was manufactured by adding a baffle 1.0 mm perpendicular to the distal opening of a 20-gauge infusion. The forces generated by the modified and standard cannulas were measured at a constant air infusion pressure of 50 mmHg by using a force sensor coupled to a schematic eye with eight radial positions from 0 degrees to 180 degrees along the circumference. Five different orientations relative to the baffle support of the modified cannula were evaluated at each position to assess the turbulent flow. RESULTS: The standard cannula produced a maximum force of 0.14 mmHg at 180 degrees from the sensor. The modified cannula generated a maximum pressure of 0.017 mmHg at 67.5 degrees from the sensor. Overall, the force from the modified cannula was an order of magnitude less than the standard infusion cannula (P < 0.01). CONCLUSIONS: The modifications of the new infusion cannula are passive with respect to its surgical performance. However, the additional baffle transforms laminar air currents into turbulent ones and allows dispersion of the jet forces, which should reduce barotrauma and desiccation of the retina during fluid-air exchange. PMID- 12652236 TI - Modified external needle drainage procedure for rhegmatogenous retinal detachment. AB - PURPOSE: To describe the results of a modified external needle subretinal fluid drainage technique to treat eyes with a rhegmatogenous retinal detachment. METHODS: One hundred eighty-seven consecutive patients with a rhegmatogenous retinal detachment who underwent scleral buckle and modified external needle drainage at Duke Eye Center or Vistar Eye Associates were included in this study. Subretinal fluid was drained by a technique whereby the intraocular pressure was raised to a supranormal level by tightening an encircling scleral buckle and then the subretinal space was entered by a needle introduced externally while the retina was directly viewed with an indirect ophthalmoscope. Subretinal fluid drainage success rate, one-operation reattachment rate, final retinal reattachment rate, intraoperative and postoperative complication rates, and final visual acuity were the main outcome measures. RESULTS: Subretinal fluid was successfully drained in all eyes. The one-operation reattachment rate was 91%, and the final reattachment rate was 98%. Intraoperatively, 15% of eyes required corneal debridement to obtain an adequate view for safe subretinal fluid drainage. Subretinal hemorrhage of no more than one clock hour occurred in 4.2% of eyes. A total of 2.6% of eyes developed an epiretinal membrane postoperatively that required a vitrectomy to improve the visual acuity. The postoperative visual acuity, 0.54 by logMAR (20/69 by Snellen), was significantly better than the preoperative visual acuity, 1.1 logMAR (20/250 by Snellen) (P < 0.0001). CONCLUSIONS: The modified external needle drainage technique is a flexible, effective method to drain subretinal fluid in eyes with a rhegmatogenous retinal detachment. The retinal reattachment rate and complication rate compare favorably to alternative subretinal fluid drainage techniques. PMID- 12652237 TI - Diagnostic and therapeutic challenges. Juxtapapillary retina capillary hemangioma. PMID- 12652238 TI - Diagnostic and therapeutic challenges. Diffuse unilateral subacute neuroretinitis. PMID- 12652239 TI - Multifocal choroiditis associated with West Nile virus encephalitis. PMID- 12652240 TI - West Nile virus chorioretinitis. PMID- 12652241 TI - Choroidal tuberculoma in miliary tuberculosis. PMID- 12652243 TI - Perfluoro-octane internal tamponade when using a temporary keratoprosthesis during retinal detachment repair. PMID- 12652242 TI - Mechanism of spontaneous closure of traumatic macular hole: OCT study of one case. PMID- 12652244 TI - Visual improvement following sequential photodynamic therapy and vitrectomy for coexisting macular hole and choroidal neovascularization. PMID- 12652245 TI - Intravitreal triamcinolone for the treatment of idiopathic juxtafoveal telangiectasis. PMID- 12652246 TI - Serpiginouslike choroiditis in a patient with cardiac sarcoidosis. PMID- 12652247 TI - Endogenous endophthalmitis secondary to vancomycin-resistant enterococci infection. PMID- 12652248 TI - Vitreoretinal degeneration complicated by retinal detachment in alport syndrome. PMID- 12652249 TI - New techniques for hemostasis during diabetic vitrectomy. PMID- 12652250 TI - Perfluorohexyloctane, pseudohypopyon, and acute glaucoma. PMID- 12652251 TI - Retinal vascular stasis associated with closed funnel stage 5 retinopathy of prematurity. PMID- 12652252 TI - Choroidal melanoma in a black patient with oculodermal melanocytosis. PMID- 12652253 TI - Diode laser photocoagulation to the ridge and avascular retina in threshold retinopathy of prematurity. PMID- 12652254 TI - Inadvertent skip areas in treatment of zone 1 retinopathy of prematurity. PMID- 12652255 TI - The dual pathway electrophysiology of the atrioventricular conduction. A new look at an old phenomenon. AB - Although we soon will be marking the 100th anniversary of the discovery of the atrioventricular (AV) node, the mysteries of this most complex of all parts of the conduction system of the heart remain. We are still battling controversies related to the precise morphology of the AV node and its atrial approaches. We are still debating the exact reentrant pathways of the AV nodal reentrant tachycardia. We are still uncertain if the so-called dual AV nodal electrophysiology encompasses two or more pathways, and what exactly makes these pathways in the absence of distinct insulated cables between the atrium and the AV node. It may be just surprising, in view of the above limitations, that current level of knowledge has nevertheless made possible some of the most spectacular successes in the modern cardiac electrophysiology. Thus, the cases of typical AVNRT are cured with a very high rate of success by radiofrequency ablations, increasing the quality of life of thousands of patients. AV nodal modifications are being performed to slow the ventricular rate during atrial fibrillation, although more progress is needed in this endeavor. The goal of the present review is to outline the major anatomic and electrophysiologic efforts in understanding the mechanisms underlying the dual pathway AV nodal propagation and to trace some novel approaches that promise to widen the horizon of the experimental and clinical fields. PMID- 12652256 TI - T wave alternans for ventricular arrhythmia risk stratification. AB - Sudden cardiac death remains one of the leading causes of death in Europe and the United States. Accordingly, the ability to identify patients at high risk of sudden cardiac death is important so that appropriate treatments can be used efficiently. Recently, T wave alternans (TWA) has emerged as a promising new test for such risk stratification. TWA is a heart rate dependent measure of arrhythmia vulnerability, with maximal predictive accuracy at sustained, regular heart rates of 100-120 bpm. In the clinical setting these conditions may be achieved by either exercise or atrial pacing. TWA has been shown to predict inducibility of ventricular tachycardia with programmed stimulation and also spontaneous arrhythmic events. TWA has been successfully applied to diverse populations, including patients with coronary artery disease, nonischemic cardiomyopathy, congestive heart failure and those with implantable defibrillators. Despite these encouraging results, the role of TWA to guide clinical therapy still needs to be better elucidated. PMID- 12652258 TI - Mitral regurgitation. AB - Mitral regurgitation is a common finding on echocardiography, seen to some degree in over 3/4 of the population. Pathologic mitral regurgitation is a common hospital diagnosis, but the percent of patients with mitral valve disease who ever require surgical correction is very small. There are many etiologies of mitral regurgitation, caused by either pathologic changes to one or more of the components of the mitral valve, including the leaflets, annulus, chordae tendineae, papillary muscles, or by abnormalities of the surrounding left ventricle and/or atrium. Mitral regurgitation can be diagnosed on physical exam or by angiography, but is best diagnosed and quantified using echocardiography. The outcome of mitral regurgitation depends on the acuity of onset of the regurgitation, as well as etiology of the mitral valve disease. Acute mitral regurgitation requires urgent mitral valve surgery. In contrast, most patients with chronic mitral regurgitation will never need corrective surgery. Currently, there is not convincing evidence that medical therapy with vasodilating medications slows the progression of mitral regurgitation. When patients with chronic mitral regurgitation develop symptoms of pathologic changes to the left ventricle, surgical treatment should be offered. Mitral valve repair is the preferred corrective surgery, and only when not possible should mitral valve replacement be performed. PMID- 12652257 TI - Use of the implantable loop recorder in patients with unexplained syncope. AB - Syncope is a complex clinical syndrome with multiple etiologies that can be very difficult to diagnose. The major obstacles to diagnosis are the periodic and unpredictable nature of events and the high spontaneous remission rate. Conventional testing is often unproductive in patients when initial non-invasive testing is negative, particularly when a paroxysmal ar-rhythmia is suspected. Holter monitoring is often employed initially, with limited utility. Holter monitoring provides a rhythm profile, but rarely achieves the gold standard of a symptom rhythm correlation. The external loop re-corder extends the period of monitoring, enhancing the diagnostic yield to as high as 60% in patients with symptoms likely to recur during a month of device use. Finally, implantable loop recorders further extend the ability to monitor cardiac patients, enhancing the diagnostic yield to as high as 85% in difficult to diagnose syncope. Several recent studies suggest that prolonged monitoring with an implantable loop recorder has a role in patients with syncope and conduction disturbances, negative tilt testing, unexplained seizures and may be superior to conventional testing with tilt and electrophysiologic studies in select patients. PMID- 12652259 TI - Peripartum cardiomyopathy. AB - Peripartum cardiomyopathy is a rare and life-threatening disease of unknown etiology. This diagnosis should be limited to previously healthy women who present with congestive heart failure (CHF) and decreased left ventricular systolic function in the last month of pregnancy or within 5 months after delivery. The diagnosis is not made in the presence of other causes of cardiac dysfunction. Patients who fail to demonstrate improvement within 2 weeks after the onset of symptoms should be evaluated for myocarditis. The type and duration of heart failure treatment is determined by the patient's heart performance at rest and with exertion. Those with normal left ventricular function at rest and with exercise or dobutamine have a good prognosis, and their medical therapy can be tapered off or discontinued over a period of 6-12 months. Patients with normal ventricular function at rest, but abnormal response to exercise should be treated for long periods of time with angiotensin converting enzyme (ACE) inhibitors or beta-blockers. Patients who continue to have depressed LV function have a poor prognosis and require treatment with appropriate medications for the rest of their lives. Pharmacological treatment includes ACE inhibitors, beta-blocking agents, diuretics, digoxin, and anticoagulation. Angiotensin converting enzyme inhibitors are used only after delivery because of their teratogenic effects. Patients who fail to recover may require inotropic therapy, intra-aortic balloon pump and left ventricular assist device as needed. Cardiac transplantation should be considered for patients who fail therapy. PMID- 12652260 TI - [Verapamil in the cardioversion of atrial fibrillation. Clinical study]. AB - BACKGROUND: Prolonged periods of atrial fibrillation (AF) or high frequency atrial pacing lead to a significant shortening of atrial refractory periods. This time-dependent electric remodelling is reduced significantly by the administration of verapamil. METHODS: The present series consists of 24 patients all suffering from atrial fibrillation (33% acute AF and 66% chronic recurrent AF) admitted to our Cardiology Department (Rome University). Group G1 (13 patients) received i.v. verapamil (50 mg in 500 cc saline solution at 40 ml/hr). Group G2 (11 patients) received amiodarone i.v. (300 mg in bolus form followed by the infusion of 900 mg in 500 cc 5% glucosate solution, 33 ml/hr for 6 hours and subsequently 18 ml/hr). All patients received non-fractionated heparin i.v. at the same time. RESULTS: No statistically significant difference was observed in the percentage of pharmacological cardioversions in the two groups: G1=61% vs G2=54% p=0.94). The patients who were not cardioverted pharmacologically were done so electrically (external DC shock). CONCLUSIONS: Albeit in this small population of patients verapamil proved to possess anti-arrhythmic effects on a par with that of standard amiodarone antiarrhythmic treatment. This antiarrhythmic potential of verapamil should be demonstrated in a broader randomised study. PMID- 12652261 TI - [Exercise-induced ST segment elevation in patients with anterior myocardial infarction: comparison with dobutamine stress]. PMID- 12652262 TI - Ventricular arrhythmias in hypertensive patients with mild metabolic alterations. AB - BACKGROUND: Ventricular arrhythmias (VA) are prevalent in hypertensives, as well as in diabetics and dyslipidemics. The aim of our study is to evaluate if alterations in glucose or lipid metabolism increase the risk of VA in hypertensive patients. METHODS: Sixty-eight patients (24 males, 44 females, aged 45-78), with mild-moderate hypertension, were divided into 2 groups: group A (n=24) normoglycemic and normolipemic, and group B (n=44) with serum cholesterol >220 mg% and/or triglycerides >165 mg% and/or glucose >110 mg%. Patients with clinical or ECG signs of ischemia or kaliemia <3.5 mEq/l, were not admitted to the study. After a 15-days pharmacological wash-out, 24-hour Holter ECG and echocardiography were performed on all patients. RESULTS: Ventricular ectopic beats (VEB) were found in 66% of group A and 100% of group B (difference =34%, 95% C.I. =18-50%). VEB were less severe in group A (Lown's class 1-2), while group B had VEB ranging from class 1 to 5. Left ventricular hypertrophy (LVH) was found in 33% of group A and in 72.7% of group B, and was not closely correlated to VA. Group B patients with LVH had more severe VA (4 A, 4 B and 5 Lown's class). CONCLUSIONS: Hyperglycemia and/or dyslipidemia, even mild, seem to increase the risk of VA in the hypertensive patient with or without LVH. It suggests that 24 hour Holter ECG may be useful to select, among patients with metabolic alterations, those at high risk for VA. PMID- 12652263 TI - [Therapeutic approach to the unilateral occlusive iliac artery disease. Preliminary results]. AB - BACKGROUND: The different therapeutic approaches to unilateral occlusive iliac artery disease are analyzed. METHODS: In the period from September 1999 to September 2001, a total of 43 patients (38 males and 5 females) has been treated for unilateral iliac artery occlusive disease. Thirty-four cases (79%) underwent an endovascular procedure, and the remaining 9 cases (21%) had a surgical intervention. Endovascular techniques included 11 cases of percutaneous transluminal angioplasty (PTA) of common iliac artery (25.6% of whole series), 5 PTA of external iliac artery (11.6%), 8 PTA+stenting of common iliac artery (18.6%) and 10 PTA+stenting of external iliac artery (23.2%). In 9 cases a surgical revascularization was performed: 6 patients underwent a femoro-femoral cross-over bypass (14%); 2 cases were treated with aorto-bifemoral reconstruction (4.7%) and one patient was operated with ilio-femoral graft (2.3 %). RESULTS: The analysis of the follow-up of our series showed, in the group of 34 patients treated with endovascular procedures, successful results were obtained in 79.4% (27 cases); in the 9 patients operated with surgical revascularization the success rate was 88.9% (8 cases ); failure rate was 20.6% for endovascular procedures and 11.1% for surgical interventions. CONCLUSIONS: The conclusion is drawn that endovascular approach (PTA, stenting) is usually the procedure of choice in the treatment of unilateral well localised lesions of the iliac artery. Conventional surgical intervention is effective for revascularizing an extensive involvement of the iliac segment or in case of bilateral disease. PMID- 12652265 TI - Thallium-201 myocardial scintigraphy in patients with Wolff-Parkinson-White syndrome. AB - Wolff-Parkinson-White syndrome (WPW) is known to cause abnormal rest electrocardiogram and stress test. Thallium-201 myocardial scintigraphy has been particularly indicated for the noninvasive evaluation of coronary artery disease in these patients. The study group consisted of 11 WPW patients with abnormal ST segment depression at rest electrocardiogram and/or stress test, with the absence of signs or symptoms of coronary artery disease. All the patients underwent exercise thallium-201 imaging associated with stress test by bicycle ergometer: 7 of them had ST-segment depression, but without other signs or symptoms of coronary artery disease. Transient and moderate myocardial perfusion defects were found in 5 of 11 patients. Perfusion defects in patients with WPW could derive from dyssynergy of ventricular activation, which could modify myocardial perfusion scintigraphy despite the absence of angiographic coronary stenosis. Previous reports and our data concluded that transient perfusion defects during exercise thallium-201 testing in WPW patients without cardiovascular disease may be observed. Thus, thallium-201 myocardial scintigraphy could present some limitations as a helpful adjunctive method for assessment of coronary artery disease in WPW patients. PMID- 12652264 TI - Recurrent varices after internal saphenectomy. Physiopathological hypothesis and clinical approach. AB - BACKGROUND: This paper analyses the causes and describes the best care of recurrent varicose veins after internal saphenectomy. METHODS: A series of 19 patients who had previously undergone internal saphenectomy were selected for surgery due to recurrent varices in the lower limbs. Clinical examination and colour duplex sonography were used as the preoperative diagnostic tools in all patients. No patients underwent phlebography. In 17 cases the main source of reflux was an incontinent saphenous stump at the level of the saphenofemoral junction with varicose cross-groin collaterals. In 2 cases recurrence was caused by incontinence of the upper thigh perforating vein. In 1 of these patients the recurrence also involved the district of the small saphenous vein. Groin neovascularisation was detected in 1 patient. RESULTS: All patients underwent groin re-dissections using transversal incisions: in 9 cases, access to the saphenofemoral junction was obtained under or at the same level as the inguinal fold, and in 10 cases using a suprainguinal route. The vertical inguinal incision was never employed. Incompetent perforating veins (thigh or leg) were ligated or sectioned in 11 patients. Ligations and exeresis of communicating veins were executed in all patients. Muller's phlebectomies were performed intra- or postoperatively on collateral varices in practically all cases. Postoperative ambulatory sclerotherapy was necessary in 6 cases. CONCLUSIONS: A correct surgical approach is only assured by diagnostic accuracy coupled with a precise hemodynamic evaluation. Correct management of the postoperative follow-up of varicose vein surgery is also important. PMID- 12652266 TI - [Endoluminal diode laser vein damage: preclinic study for vein insufficiency]. AB - BACKGROUND: The ex-vivo phase of this research aims to define fibre laser activity ranges with the intent of using a safety protocol on the operating table to ensure maximum results in occluding the vein lumen with minimum necessary energy parameters. METHODS: The study was performed on 20 human saphenous veins removed during phlebological surgery. A 1 mm diameter optical fibre was inserted in the vein to the proximal extremity where administration of laser energy began along the vein axis. The veins were subdivided into three groups based on the parameters used: Group 1: treated with 20 Watt power; Group 2, treated with 15 Watt power; Group 3, treated with 10 Watt power. On conclusion of the procedure the veins were immersed in formalin and sent for histological examination. RESULTS: The greatest endothelial damage was observed in the group of veins treated with fluence above 300 J/cm(2), in which carbonisation of the endothelium was observed; in those treated with lower fluence (200-100 J/cm(2)) damage consisted in coagulation necrosis and interstitial oedema; at fluence of 100 J/cm(2) endothelial damage was less uniform. CONCLUSIONS: Experimental evidence would appear to support the hypothesis that the optimal protocol is below 300 J/cm(2) fluence with 15 Watt power and pulse train 100-300 ms. PMID- 12652267 TI - [The role of the ambulatory care center in angiology services]. AB - Within the organization of national health care based on the DRG/ROD system, angiology services provided in "day hospital" play not only a large social role in the life of the community and the patient but also an economic one in hospital administration as there facilities allow continuation of inpatient care. Many diseases can be managed in an outpatient setting, both in diagnosis and treatment, particularly arterial hypertension, chronic obliterating arteriopathies of the lower extremities, microcirculatory and collagen disorders, VTE, and leg ulcers. A review of case records from the recent past confirms the importance of the role of the Angiology Outpatient Services plays within the Polyclinic of the University of Palermo. The exponential growth in the services provided by our facility has led to a twofold increase in the number of outpatient admissions over the past two years. This growth has produced a wider impact on the service area and allowed the introduction of angiology services that are economically profitable, while avoiding unprofitable services, through effective service management recognization. Our study results show that, where possible, outpatient care should be increasingly used. We believe that an optimal model for angiology services permits the integration of the two types of hospital care, with short stays on an inpatient basis, especially for the treatment of more severe or acute disease, and greater outpatient service utilization in ambulatory care. PMID- 12652270 TI - Mutts: baby, does your dog bite? PMID- 12652271 TI - An AED in every school: the next step for public access defibrillation. PMID- 12652272 TI - Information overload. PMID- 12652274 TI - Disorders of temperature regulation: prehospital implications. AB - Humans are warm-blooded mammals and must maintain a constant internal temperature. Body temperature is controlled by the hypothalamus in the brain, with input from sensory receptors throughout the body. At any given time, body temperature is a function of heat production and heat loss. Temperature regulation problems can result in several conditions. These include fever, hyperthermia and hypothermia. EMTs and paramedics must be familiar with the physiological and pathophysiological processes resulting from these disorders. By close observation of the patient, it should be fairly easy to determine which underlying processes are causing the observed signs and symptoms. With this knowledge, you can provide competent, compassionate prehospital care. PMID- 12652275 TI - This procedure stinks: the hazards of ammonia inhalant use. PMID- 12652276 TI - The road to med school: tips for traveling the path from EMT to physician. PMID- 12652277 TI - I didn't call for an ambulance: how to better walk the slippery slope of patient refusals. PMID- 12652278 TI - EMS Bill of Rights: what every patient deserves. AB - Every EMT, paramedic, supervisor and EMS medical director must stress that our job is to treat every patient the way we'd like to be treated or the way we'd want one of our family members treated. Those charged with supervision must ensure that EMTs and paramedics receive the support and training necessary to allow adherence to the 10 objectives outlined in this article. Quality-assurance reviews, quality-improvement programs, provider comments and complaints from the public must all be used to improve the care we deliver. Prehospital care should be constantly improving the delivery of excellent, life-saving care. PMID- 12652279 TI - Street culture: customer service in EMS. PMID- 12652280 TI - Up-front about errors: EMS system introduces proactive error reporting system. PMID- 12652281 TI - Combat hypoxia: the importance of airway management & oxygenation of the traumatic brain injury patient. AB - Brain injury, specifically TBI, is a common occurrence in our society, with devastating physical, emotional, social and economic costs. Injury not only occurs on initial impact, but also develops slowly over ensuing hours or days. TBI poses unique challenges and requires special care, especially with airway management and ventilatory treatment. Serial assessment, particularly of the GCS and pupillary changes, is vital in continuing to monitor patient status. All treatment associated with the TBI patient is aimed at restoring CPP and allowing oxygenated blood to reach injured brain tissues. PMID- 12652284 TI - In a whisper the patient began improving: new CPAP device helps patients breathe again. PMID- 12652288 TI - VPNs for EMS. PMID- 12652290 TI - Kids on calls. PMID- 12652293 TI - Alternatively spliced human tissue factor: a circulating, soluble, thrombogenic protein. AB - Tissue factor (TF) is an essential enzyme activator that forms a catalytic complex with FVII(a) and initiates coagulation by activating FIX and FX, ultimately resulting in thrombin formation. TF is found in adventitia of blood vessels and the lipid core of atherosclerotic plaques. In unstable coronary syndromes, plaque rupture initiates coagulation by exposing TF to blood. Biologically active TF has been detected in vessel walls and circulating blood. Elevated intravascular TF has been reported in diverse pro-thrombotic syndromes such as myocardial infarction, sepsis, anti-phospholipid syndrome and sickle-cell disease. It is unclear how TF circulates, although it may be present in pro coagulant microparticles. We now report identification of a form of human TF generated by alternative splicing. Our studies indicate that alternatively spliced human tissue factor (asHTF) contains most of the extracellular domain of TF but lacks a transmembrane domain and terminates with a unique peptide sequence. asHTF is soluble, circulates in blood, exhibits pro-coagulant activity when exposed to phospholipids, and is incorporated into thrombi. We propose that binding of asHTF to the edge of thrombi contributes to thrombus growth by creating a surface that both initiates and propagates coagulation. PMID- 12652294 TI - Detection of virus-specific T cells and CD8+ T-cell epitopes by acquisition of peptide-HLA-GFP complexes: analysis of T-cell phenotype and function in chronic viral infections. AB - Antigen-specific CD8+ T cells acquire peptide-major histocompatibility complex (MHC) clusters through T-cell receptor (TCR)-mediated endocytosis after specific antigen stimulation. We generated an antigen-presenting cell (APC) expressing human leukocyte antigen (HLA)-A*201 coupled to the enhanced green fluorescent protein (GFP), which delivered GFP to an antigen-specific T cell when pulsed with antigenic peptide. We quantitatively identified human T-cell lymphotropic virus type I (HTLV-I) Tax(11-19) peptide-specific T-cell populations in peripheral blood mononuclear cells (PBMCs) from patients with HTLV-I-associated neurologic disease and defined a new CD8+ T-cell epitope in the HTLV-I envelope region. Acquisition of peptide-HLA-GFP complexes by antigen-specific T cells could distinguish, with respect to phenotype and perforin production, T cells from the chronic viral infections cytomegalovirus and HTLV-I. This approach will be a powerful tool in understanding the role of antigen-specific T-cell responses in health and disease. PMID- 12652295 TI - A novel function for tissue inhibitor of metalloproteinases-3 (TIMP3): inhibition of angiogenesis by blockage of VEGF binding to VEGF receptor-2. AB - Tissue inhibitor of metalloproteinases-3 (TIMP3) is one of four members of a family of proteins that were originally classified according to their ability to inhibit matrix metalloproteinases (MMP). TIMP3, which encodes a potent angiogenesis inhibitor, is mutated in Sorsby fundus dystrophy, a macular degenerative disease with submacular choroidal neovascularization. In this study we demonstrate the ability of TIMP3 to inhibit vascular endothelial factor (VEGF) mediated angiogenesis and identify the potential mechanism by which this occurs: TIMP3 blocks the binding of VEGF to VEGF receptor-2 and inhibits downstream signaling and angiogenesis. This property seems to be independent of its MMP inhibitory activity, indicating a new function for this molecule. PMID- 12652296 TI - SKAP-55 regulates integrin adhesion and formation of T cell-APC conjugates. AB - Src kinase-associated phosphoprotein of 55 kDa (SKAP-55; encoded by SCAP1) is a T cell adaptor protein of unknown function that contains a pleckstrin homology and an SH3 domain. Here we show that SKAP-55 regulates integrin-mediated adhesion and conjugate formation between T cells and antigen-presenting cells (APCs). SKAP-55 enhances adhesion to fibronectin and intercellular adhesion molecule-1 (ICAM-1), colocalizes with actin at the T cell-APC synapse and promotes the clustering of lymphocyte-associated antigen-1 (LFA-1). Enhanced conjugation is comparable to that induced by adhesion and degranulation-promoting adaptor protein (ADAP), a binding partner of SKAP-55, and is abrogated by deletion of the SKAP-55 SH3 domain. Conjugate formation is accompanied by the translocation of SKAP-55 to membrane rafts, an event that is regulated by both LFA-1 and T cell receptor ligation. Our findings identify a mechanism by which SKAP-55 modulates T cell responses to antigen. PMID- 12652297 TI - Caspase inhibition causes hyperacute tumor necrosis factor-induced shock via oxidative stress and phospholipase A2. AB - Dysregulated apoptotic cell death contributes to many pathological conditions, including sepsis, prompting the suggestion that caspase inhibition to block apoptosis could have useful therapeutic applications. Because the cytokine tumor necrosis factor (TNF, also known as TNF-alpha) is both pro-apoptotic and pro inflammatory and is involved in septic shock, we tested whether caspase inhibition would alleviate TNF-induced toxicity in vivo. General caspase inhibition by the protease inhibitor zVAD-fmk exacerbated TNF toxicity by enhancing oxidative stress and mitochondrial damage, resulting in hyperacute hemodynamic collapse, kidney failure and death. Thus, survival of TNF toxicity depends on caspase-dependent processes. Our results demonstrated the pathophysiological relevance of caspase-independent, ROS-mediated pathways in response to lethal TNF-induced shock in mice. In addition, survival of TNF toxicity seemed to require a caspase-dependent protective feedback on excessive reactive oxygen species (ROS) formation and phospholipase A2 activation. PMID- 12652298 TI - Mutations in RAI1 associated with Smith-Magenis syndrome. AB - Smith-Magenis syndrome (SMS) is a mental retardation syndrome associated with deletions involving chromosome 17p11.2. Persons with SMS have characteristic behavioral abnormalities, including self-injurious behaviors and sleep disturbance, and distinct craniofacial and skeletal anomalies. We identified dominant frameshift mutations leading to protein truncation in RAI1 in three individuals who have phenotypic features consistent with SMS but do not have 17p11.2 deletions detectable by standard fluorescence in situ hybridization techniques. PMID- 12652300 TI - Additional SNPs and linkage-disequilibrium analyses are necessary for whole genome association studies in humans. AB - More than 5 million single-nucleotide polymorphisms (SNPs) with minor-allele frequency greater than 10% are expected to exist in the human genome. Some of these SNPs may be associated with risk of developing common diseases. To assess the power of currently available SNPs to detect such associations, we resequenced 50 genes in two ethnic samples and measured patterns of linkage disequilibrium between the subset of SNPs reported in dbSNP and the complete set of common SNPs. Our results suggest that using all 2.7 million SNPs currently in the database would detect nearly 80% of all common SNPs in European populations but only 50% of those common in the African American population and that efficient selection of a minimal subset of SNPs for use in association studies requires measurement of allele frequency and linkage disequilibrium relationships for all SNPs in dbSNP. PMID- 12652299 TI - Erosion of the telomeric single-strand overhang at replicative senescence. AB - Cultured primary human cells inevitably enter a state of replicative senescence for which the specific molecular trigger is unknown. We show that the single strand telomeric overhang, a key component of telomere structure, is eroded at senescence. Expression of telomerase prevents overhang loss, suggesting that this enzyme prevents senescence by maintaining proper telomere structure. In contrast, progressive overhang loss occurs in cells that avoid senescence through the inactivation of p53 and Rb, indicating that overhang erosion is the result of continuous cell division and not a consequence of senescence. We thus provide evidence for a specific molecular alteration in telomere structure at senescence and suggest that this change, rather than overall telomere length, serves to trigger this state. PMID- 12652301 TI - Quality and completeness of SNP databases. AB - To address the quality and completeness of single-nucleotide polymorphism (SNP) databases, we resequenced 173 kb (spanning 17 loci) in 150 chromosomes of west African and European ancestry. Over 88% of SNPs in the public (TSC and BAC overlap) and Celera databases were confirmed in independent resequencing. Approximately 45% of all human heterozygosity is attributable to SNPs already available from the two databases, and of SNPs with minor-allele frequencies >10%, more than half are represented. PMID- 12652302 TI - A defective response to Hedgehog signaling in disorders of cholesterol biosynthesis. AB - Smith-Lemli-Opitz syndrome (SLOS), desmosterolosis and lathosterolosis are human syndromes caused by defects in the final stages of cholesterol biosynthesis. Many of the developmental malformations in these syndromes occur in tissues and structures whose embryonic patterning depends on signaling by the Hedgehog (Hh) family of secreted proteins. Here we report that response to the Hh signal is compromised in mutant cells from mouse models of SLOS and lathosterolosis and in normal cells pharmacologically depleted of sterols. We show that decreasing levels of cellular sterols correlate with diminishing responsiveness to the Hh signal. This diminished response occurs at sterol levels sufficient for normal autoprocessing of Hh protein, which requires cholesterol as cofactor and covalent adduct. We further find that sterol depletion affects the activity of Smoothened (Smo), an essential component of the Hh signal transduction apparatus. PMID- 12652304 TI - Reading skills are related to global, but not local, acoustic pattern perception. PMID- 12652303 TI - Processing of low-probability sounds by cortical neurons. AB - The ability to detect rare auditory events can be critical for survival. We report here that neurons in cat primary auditory cortex (A1) responded more strongly to a rarely presented sound than to the same sound when it was common. For the rare stimuli, we used both frequency and amplitude deviants. Moreover, some A1 neurons showed hyperacuity for frequency deviants--a frequency resolution one order of magnitude better than receptive field widths in A1. In contrast, auditory thalamic neurons were insensitive to the probability of frequency deviants. These phenomena resulted from stimulus-specific adaptation in A1, which may be a single-neuron correlate of an extensively studied cortical potential- mismatch negativity--that is evoked by rare sounds. Our results thus indicate that A1 neurons, in addition to processing the acoustic features of sounds, may also be involved in sensory memory and novelty detection. PMID- 12652305 TI - Single auditory neurons rapidly discriminate conspecific communication signals. PMID- 12652306 TI - Otx dose-dependent integrated control of antero-posterior and dorso-ventral patterning of midbrain. AB - Organizing centers emit signaling molecules that specify different neuronal cell types at precise positions along the anterior-posterior (A-P) and dorsal-ventral (D-V) axes of neural tube during development. Here we report that reduction in Otx proteins near the alar-basal plate boundary (ABB) of murine midbrain resulted in a dorsal shift of Shh expression, and reduction in Otx proteins at the midbrain-hindbrain boundary (MHB) resulted in an anterior expansion of the Fgf8 domain. Our data thus indicate that an Otx dose-dependent repressive effect coordinates proper positioning of Shh and Fgf8 expression. Furthermore, this control is effective for conferring proper cell identity in the floor-plate region of midbrain and does not require an Otx2-specific property. We propose that this mechanism may provide both A-P and D-V positional information to neuronal precursors located within the midbrain. PMID- 12652307 TI - A new link between the c-Abl tyrosine kinase and phosphoinositide signalling through PLC-gamma1. AB - The c-Abl tyrosine (Tyr) kinase is activated after platelet-derived-growth factor receptor (PDGFR) stimulation in a manner that is partially dependent on Src kinase activity. However, the activity of Src kinases alone is not sufficient for activation of c-Abl by PDGFR. Here we show that functional phospholipase C-gamma1 (PLC-gamma1) is required for c-Abl activation by PDGFR. Decreasing cellular levels of phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2) by PLC-gamma1 mediated hydrolysis or dephosphorylation by an inositol polyphosphate 5 phosphatase (Inp54) results in increased Abl kinase activity. c-Abl functions downstream of PLC-gamma1, as expression of kinase-inactive c-Abl blocks PLC gamma1-induced chemotaxis towards PDGF-BB. PLC-gamma1 and c-Abl form a complex in cells that is enhanced by PDGF stimulation. After activation, c-Abl phosphorylates PLC-gamma1 and negatively modulates its function in vivo. These findings uncover a newly discovered functional interdependence between non receptor Tyr kinase and lipid signalling pathways. PMID- 12652308 TI - Ku70 suppresses the apoptotic translocation of Bax to mitochondria. AB - Bax induces mitochondrial-dependent cell death signals in mammalian cells. However, the mechanism of how Bax is kept inactive has remained unclear. Yeast based functional screening of Bax inhibitors from mammalian cDNA libraries identified Ku70 as a new Bax suppressor. Bax-mediated apoptosis was suppressed by overexpression of Ku70 in mammalian cells, but enhanced by downregulation of Ku70. We found that Ku70 interacts with Bax, and that the carboxyl terminus of Ku70 and the amino terminus of Bax are required for this interaction. Bax is known to translocate from the cytosol to mitochondria when cells receive apoptotic stimuli. We found that Ku70 blocks the mitochondrial translocation of Bax. These results suggest that in addition to its previously recognized DNA repair activity in the nucleus, Ku70 has a cytoprotective function in the cytosol that controls the localization of Bax. PMID- 12652309 TI - Cytoprotective membrane-permeable peptides designed from the Bax-binding domain of Ku70. AB - Bax is a pro-apoptotic member of Bcl-2 family proteins and is central to mitochondria-dependent apoptosis. Bax resides in the cytosol as a quiescent protein and translocates into mitochondria after apoptotic stimuli. Ku70 is a 70K subunit of the Ku complex, which has an important role in DNA double-strand break (DSB) repair in the nucleus. In another article in this issue, we reported that Ku70 interacts with pro-apoptotic protein Bax in the cytosol and prevents its mitochondrial translocation, suggesting that Ku70 suppresses Bax-mediated apoptosis. Here, we describe the development of a new membrane-permeable peptide, Bax-inhibiting peptide (BIP) that inhibits Bax-mediated apoptosis, on the basis of the previous finding that showed an interaction between Ku70 and Bax. BIP is comprised of five amino acids designed from the Bax-binding domain of Ku70, and suppresses the mitochondrial translocation of Bax. BIP inhibited Bax-mediated apoptosis induced by staurosporine, UVC irradiation and anti-cancer drugs in several types of cells. BIP may provide valuable information in the development of therapeutics that control apoptosis-related diseases. PMID- 12652310 TI - Exocytosis of single chromaffin granules in cell-free inside-out membrane patches. AB - In chromaffin cells, exocytosis of single granules and properties of the fusion pore--the first connection between vesicular lumen and extracellular space --can be studied by cell-attached patch amperometry, which couples patch-clamp capacitance measurements with simultaneous amperometric recordings of transmitter release. Here we have studied exocytosis of single chromaffin granules and endocytosis of single vesicles in cell-free inside-out membrane patches by patch capacitance measurements and patch amperometry. We excised patches from chromaffin cells by using methods developed for studying properties of single ion channels. With low calcium concentrations in the pipette and bath, the patches showed no spontaneous exocytosis, but exocytosis could be induced in some patches by applying calcium to the cytoplasmic side of the patch. Exocytosis was also stimulated by calcium entry through the patch membrane. Initial conductances of the fusion pore were undistinguishable in cell-attached and excised patch recordings, but the subsequent pore expansion was slower in excised patches. The properties of exocytotic fusion pores in chromaffin cells are very similar to those observed in mast cells and granulocytes. Excised patches provide a tool with which to study the mechanisms of fusion pore formation and endocytosis in vitro. PMID- 12652312 TI - Immunotargeting of catalase to the pulmonary endothelium alleviates oxidative stress and reduces acute lung transplantation injury. AB - Vascular immunotargeting may facilitate the rapid and specific delivery of therapeutic agents to endothelial cells. We investigated whether targeting of an antioxidant enzyme, catalase, to the pulmonary endothelium alleviates oxidative stress in an in vivo model of lung transplantation. Intravenously injected enzymes, conjugated with an antibody to platelet-endothelial cell adhesion molecule-1, accumulate in the pulmonary vasculature and retain their activity during prolonged cold storage and transplantation. Immunotargeting of catalase to donor rats augments the antioxidant capacity of the pulmonary endothelium, reduces oxidative stress, ameliorates ischemia-reperfusion injury, prolongs the acceptable cold ischemia period of lung grafts, and improves the function of transplanted lung grafts. These findings validate the therapeutic potential of vascular immunotargeting as a drug delivery strategy to reduce endothelial injury. Potential applications of this strategy include improving the outcome of clinical lung transplantation and treating a wide variety of endothelial disorders. PMID- 12652311 TI - Serial in vivo imaging of the targeted migration of human HSV-TK-transduced antigen-specific lymphocytes. AB - New technologies are needed to characterize the migration, survival, and function of antigen-specific T cells in vivo. Here, we demonstrate that Epstein-Barr virus (EBV)--specific T cells transduced with vectors encoding herpes simplex virus-1 thymidine kinase (HSV-TK) selectively accumulate radiolabeled 2'-fluoro-2'-deoxy 1-beta-D-arabinofuranosyl-5-iodouracil (FIAU). After adoptive transfer, HSV-TK+ T cells labeled in vitro or in vivo with [131I]FIAU or [124I]FIAU can be noninvasively tracked in SCID mice bearing human tumor xenografts by serial images obtained by scintigraphy or positron emission tomography (PET), respectively. These T cells selectively accumulate in EBV+ tumors expressing the T cells' restricting HLA allele but not in EBV- or HLA-mismatched tumors. The concentrations of transduced T cells detected in tumors and tissues are closely correlated with the concentrations of label retained at each site. Radiolabeled transduced T cells retain their capacity to eliminate targeted tumors selectively. This technique for imaging the migration of ex vivo-transduced antigen-specific T cells in vivo is informative, nontoxic, and potentially applicable to humans. PMID- 12652322 TI - Substrate-induced transmembrane signaling in the cobalamin transporter BtuB. AB - The outer membranes of Gram-negative bacteria possess transport proteins essential for uptake of scarce nutrients. In TonB-dependent transporters, a conserved sequence of seven residues, the Ton box, faces the periplasm and interacts with the inner membrane TonB protein to energize an active transport cycle. A critical mechanistic step is the structural change in the Ton box of the transporter upon substrate binding; this essential transmembrane signaling event increases the affinity of the transporter for TonB and enables active transport to proceed. We have solved crystal structures of BtuB, the outer membrane cobalamin transporter from Escherichia coli, in the absence and presence of cyanocobalamin (vitamin B(12)). In these structures, the Ton box is ordered and undergoes a conformational change in the presence of bound substrate. Calcium has been implicated as a necessary factor for the high-affinity binding (K(d) approximately 0.3 nM) of cyanocobalamin to BtuB. We observe two bound calcium ions that order three extracellular loops of BtuB, thus providing a direct (and unusual) structural role for calcium. PMID- 12652323 TI - Ventricular rate control in atrial fibrillation: what is the optimal rate? The concept of controlling the heart rate burden. PMID- 12652324 TI - Ventricular fibrillation resulting from acute right ventricular infarction from isolated occlusion of a right ventricular branch artery. AB - Malignant ventricular arrhythmias can result from isolated right ventricular infarction, and reports of this phenomenon in the literature are rare. We present a case of a 46-year-old man with acute onset of chest pain angiographically confirmed to be a result of isolated occlusion of a right ventricular branch artery. He developed ventricular fibrillation within 5 hours of symptom onset. This case highlights the point that despite its benign clinical appearance and preserved left ventricular function, necrosis of right ventricular tissue can have life-threatening consequences. PMID- 12652325 TI - Bivalirudin (Angiomax) use during intracoronary brachytherapy may predispose to acute closure. AB - We describe two cases of intracoronary vascular brachytherapy where bivalirudin (Angiomax), employed as an anticoagulant, led to abrupt vessel closure or threatened abrupt closure. Use of bivalirudin (Angiomax) during intracoronary brachytherapy may predispose to the formation of intracoronary thrombus, related to the reversible binding kinetics of the bivalirudin to thrombin, and resulting in recovery of thrombin functional activity during periods of prolonged stasis that occur during intracoronary brachytherapy. Intracoronary abciximab administration may be a useful strategy in resolving the acute closure, since abciximab administered early during the formation of thrombus has been shown to facilitate clot lysis. PMID- 12652326 TI - Left ventricular performance during acute rate control in atrial fibrillation: the importance of heart rate and agent used. AB - BACKGROUND: The relation between heart rate and left ventricular function during rate control in atrial fibrillation is incompletely understood. METHODS: Twenty four patients (age 67 +/- 11 years) with symptomatic recent onset rapid atrial fibrillation and rapid ventricular rate (> 110 bpm) were randomly assigned to receive either intravenous digoxin (13 mcg/kg) or intravenous diltiazem (0.25 mg/kg bolus plus a maintenance infusion). A portable radionuclide detector was used to collect validated measures of relative left ventricular volumes, along with heart rate data, every 15 seconds for 6 hours. RESULTS: Heart rate decreased significantly at 15 minutes and 180 minutes in the diltiazem group (from 133 +/- 18 bpm to 111 +/- 26 bpm [P <.01] to 94 +/- 24 bpm [P <.001]) but not in the digoxin group (from 129 +/- 18 bpm to 126 +/- 17 bpm [P = NS] to 118 +/- 15 bpm [P = NS]). Left ventricular ejection fraction improved in both groups to a similar extent (from 39 +/- 10% to 50 +/- 8%, [P <.05] after diltiazem, and from 38 +/- 8% to 52 +/- 11% [P <.05] after digoxin at baseline vs 180 minutes, respectively). The ejection fraction vs heart rate slope was steeper in the digoxin group than in the diltiazem group (-0.34 +/- 0.18 vs -0.16 +/- 0.17, P =.048) indicating a more pronounced improvement in ejection fraction per unit decrease in heart rate. CONCLUSION: In patients with acute atrial fibrillation, digoxin led to similar improvements in ejection fraction compared to diltiazem despite a slower and less potent heart rate slowing. PMID- 12652327 TI - Correlation of Angiotensin-converting enzyme gene polymorphism with effect of antihypertensive therapy by Angiotensin-converting enzyme inhibitor. AB - BACKGROUND: The correlation of the angiotensin-converting enzyme insertion/deletion polymorphism with essential hypertension shown in previous studies is controversial, and the responses of hypertensive patients with different angiotensin-converting enzyme genotypes to treatment with an angiotensin-converting enzyme inhibitor merits investigation. METHODS: Eighty nine patients with essential hypertension and 102 normotensive subjects were included in this study. All subjects were genotyped by polymerase chain reaction for the insertion/deletion polymorphism of the angiotensin-converting enzyme gene. Blood pressure was measured before and after taking Benazepril 10 mg, once daily, for 2 months. RESULTS: The genotype and allele frequencies were similar (chi(2) = 0.64, P =.73; chi(2) = 0.36, P =.55, respectively). The reduction of both systolic and diastolic blood pressure in the DD genotype was significantly greater than in the II genotype (10.13 +/- 4.91 vs 5.37 +/- 2.79, P <.01; 7.47 +/ 3.50 vs 4.71 +/- 2.40, P <.05, respectively). CONCLUSIONS: No significant association of angiotensin-converting enzyme gene polymorphism with essential hypertension was found. Angiotensin-converting enzyme gene polymorphism might be related to the antihypertensive response to an angiotensin-converting enzyme inhibitor in hypertensive patients. PMID- 12652328 TI - Comparison of the effect of class IA antiarrhythmic drugs on transmembrane potassium currents in rabbit ventricular myocytes. AB - BACKGROUND: The blockade of cardiac transmembrane potassium channels, which is commonly seen with various antiarrhythmic drugs, plays an important role in their mechanism of action. We studied and compared the less-explored effects of three Class IA antiarrhythmics on the transient outward current (I(to)) and on the inward rectifier (I(kl)), ATP sensitive (I(KATP)), and delayed rectifier (I(K)) potassium currents in rabbit ventricular myocytes. METHODS AND RESULTS: Transmembrane currents were measured by applying the whole-cell configuration of the patch-clamp technique at 37 degrees C in myocytes enzymatically isolated from rabbit ventricular preparations. Quinidine (10 microM), disopyramide (10 microM), and procainamide (50 microM) were studied at concentrations close to or exceeding the therapeutic plasma level. All studied drugs significantly decreased the amplitude of I(KATP) (activated by 50 microM pinacidil) and I(K) currents. None of them influenced significantly I(kl). The amplitude of I(to) was decreased by quinidine and disopyramide but was not considerably altered by procainamide. The fast inactivation of I(to) was not changed by procainamide and was significantly accelerated by quinidine and disopyramide. CONCLUSION: Although quinidine, disopyramide, and procainamide are all classified as Class IA antiarrhythmics, these drugs had different effects on various potassium currents, which may partially explain their distinct effect on repolarization in various cardiac tissues and on cardiac arrhythmias in clinical settings. PMID- 12652329 TI - Effect of acute intravenous cocaine administration on endothelium-dependent vasodepressor responses to acetylcholine. AB - BACKGROUND: Cardiovascular events commonly associated with acute and chronic cocaine abuse include coronary vasospasm, arrhythmias, myocardial infarction, and sudden death. It has been suggested that cocaine causes endothelial dysfunction and vasoconstriction by inhibiting local production of nitric oxide and that endothelial dysfunction may be involved in the cardiovascular events associated with cocaine use. The present study investigated the effect of acute intravenous cocaine administration on endothelium-dependent vasodepressor responses to acetylcholine in the anesthetized rat. METHODS AND RESULTS: Rats were anesthetized with intraperitoneally Inactin (140 mg/kg) and catheters were inserted into the jugular vein and iliac artery for the injection of drugs and measurement of systemic arterial pressure. A thermistor catheter was advanced to the aortic arch for the measurement of cardiac output via the thermal dilution technique. A 5 mg/kg intravenous dose of cocaine, which enhances the vasopressor response to norepinephrine and blocks the vasopressor response to tyramine, had no effect on the vasodepressor response to the endothelium-dependent vasodilator acetylcholine. In addition, responses to the nitric oxide-donor sodium nitroprusside and the vasoconstrictor peptide angiotensin II were not altered by administration of cocaine. CONCLUSIONS: Acute administration of cocaine in a dose that blocks norepinephrine uptake into adrenergic terminals had no effect on endothelial function as measured by the absence of an effect on the vasodepressor response to intravenous injections of acetylcholine. Although chronic cocaine exposure has been shown to cause vascular dysfunction, eventually leading to impairment of the nitric oxide pathway, it appears that acute cocaine administration does not inhibit this pathway in the anesthetized rat. PMID- 12652330 TI - Separate and combined effects of local and continuous intravenous administration of beta-cyclodextrin tetradecasulfate on intimal hyperplasia after angioplasty in porcine coronary arteries. AB - BACKGROUND: Beta-Cyclodextrin tetradecasulfate binds fibroblast growth factors and possesses anticoagulant properties. This study was designed to assess the separate and combined effects of local intramural delivery and intravenous administration of beta-cyclodextrin tetrade-casulfate on neointimal formation and arterial damage following angioplasty. METHODS AND RESULTS: Fifty-two pigs randomized into four groups underwent coronary artery angioplasty: 1) control, 2) continuous intravenous infusion of 100 mg/kg/d of beta-cyclodextrin tetradecasulfate, 3) intramural delivery of 1250 mg beta-cyclodextrin tetradecasulfate, 4) intramural delivery of 1250 mg beta-cyclodextrin tetradecasulfate followed by continuous intravenous infusion of 100 mg/kg/d. Fourteen days after injury, morphometric analysis revealed that arteries randomized to the intravenous beta-cyclodextrin tetradecasulfate groups had a decreased normalized neointima area: control, 3.03 +/- 0.75 mm(2); intravenous, 1.67 +/- 0.73 mm(2) (40% decrease; P < 10(-7)); intravenous plus local, 1.95 +/- 0.76 mm(2) (30% decrease; P < 10(-5)). There was no difference in neointimal response following local beta-cyclodextrin tetradecasulfate delivery only (2.82 +/- 1.14 mm(2)). Coronary arterial damage, defined as aneurysm, dissection, adventitial rupture, and retromedial hematoma was similar in all groups (12% in control and local groups, 10% in the intravenous group, 14% in the intravenous plus local; NS). Bleeding complications were more frequent in the intravenous and intravenous plus local groups compared to the local and control groups (23%vs 7.6% and 0%, respectively; P < 0.05). CONCLUSIONS: Continuous intravenous administration of beta-cyclodextrin tetradecasulfate substantially reduced intimal hyperplasia, while intramural delivery had no effect, indicating that a single bolus of beta-cyclodextrin tetradecasulfate did not reduce intimal hyperplasia. There was no additive effect of local intramural delivery of beta cyclodextrin tetradecasulfate. However, local delivery of beta-cyclodextrin tetradecasulfate induced less bleeding complications and did not lead to additional arterial injury, indicating that local delivery of an anticoagulant does not cause additional arterial injury. PMID- 12652331 TI - Suppression of oxidative stress as a mechanism of reduction of hypercholesterolemic atherosclerosis by aspirin. AB - BACKGROUND: Hypercholesterolemia increases the formation of arachidonic acid and thereby synthesis of prostaglandins and leukotrienes. During synthesis of these eicosanoids, oxyradicals are produced. Oxyradicals have been implicated in the development of hypercholes-terolemic atherosclerosis. Aspirin, an inhibitor of synthesis of prostaglandins, would prevent the generation of oxyradicals and hence would prevent the development of atherosclerosis. The purpose of the investigation was to determine if aspirin attenuates the development of hypercholesterolemic atherosclerosis, and if this attenuation is associated with a decrease in the oxidative stress. METHODS AND RESULTS: Three groups of rabbits were used for this study: Group I, control; Group II, 0.5% cholesterol; Group III, 0.5% cholesterol plus 0.068% aspirin. Blood samples were collected before and after 1 and 2 months of experimental diets for measurement of serum triglycerides, total cholesterol, low-density lipoprotein cholesterol, high density lipoprotein cholesterol, serum malondialdehyde, and white blood cell chemiluminescence, a measure of oxyradicals produced by white blood cells. Aortas were removed at the end of the protocol for the measurement of atherosclerotic plaques, malondialdehyde and aortic chemiluminescence, a measure of antioxidant reserve. Serum total cholesterol, low-density lipoprotein cholesterol, high density lipoprotein cholesterol, and ratio of total cholesterol to high-density lipoprotein cholesterol increased to a similar extent in Groups II and III compared to Group I. Serum triglycerides increased in both Groups II and III; however, the increase was greater in Group III than in Group II. Levels of serum and aortic malondialdehyde, and white blood cell-chemiluminescence were higher in Group II compared to Group I. Aspirin decreased the levels of these parameters. Antioxidant reserve increased in both Groups II and III, the increase being greater in the latter than the former. White blood cell-chemiluminescence increased in Group II compared to Group I, but remained unaltered in Group III compared to Group I. Aspirin treatment reduced the development of atherosclerosis by approximately 47%. CONCLUSION: These results suggest that aspirin, an inhibitor of cyclooxygenase, reduced the development of hypercholesterolemic atherosclerosis, and this effect was associated with a decrease in the oxidative stress. PMID- 12652332 TI - Cardiovascular pharmacogenomics: current status, future prospects. AB - Pharmacogenomics is a field dedicated to exploring the contribution of genetics to interindividual variability in drug response. A goal of cardiovascular pharmacogenomics is to guide cardiovascular drug development and selection so as to optimize therapeutic benefit and minimize the potential for toxicity. Genetic based differences in drug metabolism have long been recognized but just now are on the verge of wider clinical application. Differences in efficacy of cardiovascular drugs (independent of drug concentration) based on common genetic variations (polymorphisms) only recently have begun to be explored, but the potential for clinical application appears promising. Examples are presented of important pharmacodynamic effects of genetic variants on several drugs, including those in antiarrhythmic, reninangiotensin, beta-blocker, lipid-lowering, and antithrombotic classes. Principles of pharmacogenomics applied to drug metabolism are discussed that are relevant to drug development and clinical use, and examples are given for CYP450 phase I enzymes, phase II enzymes, and drug transporters. Challenges in establishing true pharmacogenetic associations are discussed, and current and future clinical potential is summarized. Rapid research progress and initial clinical applications with pharmacogenomics are foreseen in the near future. PMID- 12652333 TI - Side-to-side symmetry of radioprotein transfer from tissue space to systemic vasculature following subcutaneous injection in normal subjects and patients with breast cancer. AB - Quantitative lymphoscintigraphy can be used for investigation of unilateral lymphatic disease of the limbs, such as breast cancer-related lymphoedema (BCRL). Previous studies have compared lymphatic function in the affected limb with that in the unaffected contralateral limb. This study aims to confirm that the assumption of pre-morbid symmetry, never previously demonstrated, is valid. A dual-isotope technique, with bilateral subcutaneous hand injection of polyclonal human immunoglobulin G (HIgG) labelled with either technetium-99m or indium-111, was performed on a total of 37 subjects. The use of two different labels, one for each limb, enabled comparison not only of the rate of clearance from the injection depot, but also of the rate of appearance in venous blood. Results demonstrate clear symmetry between the two arms with respect to both depot clearance and blood appearance rates, as well as the coupling between these two variables. In unilateral lymphatic disease, results of quantitative lymphoscintigraphy should be expressed in relation to the normal arm rather than to an independent control population. PMID- 12652334 TI - MR imaging of cartilage repair procedures. AB - It is becoming increasingly important for the radiologist to evaluate the appearance and outcome of cartilage repair procedures. MR imaging is currently the best method for such evaluation but it is necessary to use cartilage-specific sequences and to modify those sequences when necessary to minimize artifacts from retained metal within the joint. This article reviews the surgical technique of the more commonly performed cartilage repair procedures, currently recommended techniques for the MR imaging evaluation of articular cartilage and cartilage repair procedures, and the MR imaging appearance of cartilage repair procedures and of the most frequently encountered complications following such procedures. PMID- 12652335 TI - 18FDG PET scanning of benign and malignant musculoskeletal lesions. AB - OBJECTIVE: To describe the technique, applications and advantages of (18)FDG PET scanning in detection, analysis and management of musculoskeletal lesions. DESIGN AND PATIENTS: Forty-five patients (19 males,26 females) aged 9 to 81 years had radiographs, routine radionuclide scans, CT and/or MRI of clinically suspected active benign or malignant musculoskeletal lesions. (18)FDG scans with a Siemens ECAT EXACT 921 dedicated PET unit (Knoxville, Tenn.) and FWH=6 mm images acquired as a 5-6 bed examination (6 min emission and 4 min transmission) used OSEM iterative reconstruction with segmented transmission attenuation correction and a Gaussian filter (cutoff 6.7 mm). Region of interest (ROI) 3x3 pixel image analysis based on transverse whole body images (slice thickness 3.37 mm) generated Maximum Standard Uptake Values (Max SUV) with a cutoff of 2.0 used to distinguish benign and malignant lesions. RESULTS: Thirty-nine studies were available for SUV ROI analysis. Overall sensitivity for differentiating malignant from benign osseous and non-osseous lesions was 91.7% (22/24), overall specificity was 100% (11/11) with an accuracy of 91.7%. All aggressive lesions had a Max SUV >2.0. Data separating benign from malignant lesions and aggressive from benign lesions were statistically significant ( P<0.001) in both categories. There was no statistically significant difference in distinguishing aggressive from malignant lesions ( P, ns). CONCLUSION: (18)FDG PET contributes unique information regarding metabolism of musculoskeletal lesions. By supplying a physiologic basis for more informed treatment and management, it influences prognosis and survival. Moreover, since residual, recurrent or metastatic tumors can be simultaneously documented on a single whole body scan, PET may theoretically prove to be cost-effective. PMID- 12652337 TI - Desmoid tumor of bone with enchondromatous nodules, mistaken for chondrosarcoma. AB - Desmoid tumor of bone, also termed desmoplastic fibroma or aggressive fibromatosis, is a rare, locally aggressive fibroblastic tumor. We present a 16 year-old male with a huge desmoid tumor involving the iliac wing. It was associated with enchondromatous nodules mimicking malignancy. The tumor in this patient was mistaken for chondrosarcoma and hemipelvectomy was performed. To our knowledge, such a case has not previously been documented fully in the English literature. The radiographic and pathologic findings and a possible mechanism of enchondromatous nodule formation in fibrous bone tumors are discussed. PMID- 12652336 TI - Intraosseous lipoma: report of 35 new cases and a review of the literature. AB - OBJECTIVE: To identify the common imaging features of intraosseous lipomas on radiographs, magnetic resonance imaging (MRI) and computed tomography (CT), and review their histological features. DESIGN AND PATIENTS: Thirty-five previously unreported cases of intraosseous lipoma were reviewed and a meta-analysis was performed of another 110 cases identified from the English language literature. RESULTS: The mean age at presentation is 43 years. Sex distribution is nearly equal. Lipomas occur most frequently in the lower limb (71% overall), particularly in the os calcis (32%). Other common sites include the metaphyses of long bones, where lesions are typically eccentric. Lipomas are usually well defined, but marginal sclerosis is commoner in lesions of the os calcis (61%) than at other sites (38%). Calcification is also more frequent in the os calcis (62%), and almost invariably centrally located. Calcification at other sites is less common (30%), and is more variable in appearance. Bone expansion is less common (30%), and usually minimal. Fat necrosis and cyst formation identified on MRI is common (67%), and more frequent in the os calcis. CONCLUSIONS: Although there is correlation between the histological and radiological features of intraosseous lipomas in general, some discrepancies occur in the radiological appearances of lipomas in different sites. The evidence that these lesions are true benign tumours of fat is controversial. Several aetiological factors have been implicated in their development. The constant location of os calcis lesions at the critical angle suggests an aetiology that may be related to biomechanical lines of stress. In other instances it is possible that involution of pre existing lesions may lead to the development of lipomas. PMID- 12652338 TI - A rare case of infantile Ollier's disease demonstrating bilaterally symmetric extremity involvement. AB - We report an unusual case of Ollier's disease presenting at an atypically early age and with extensive, bilaterally symmetric features. PMID- 12652339 TI - Primary periosteal lymphoma: an unusual presentation of non-Hodgkin's lymphoma with radiographic, MR imaging, and pathologic correlation. AB - This report describes a primary periosteal location of non-Hodgkin's lymphoma, without nodal disease, and without adjacent intramedullary disease at presentation. The clinical and imaging appearance of periosteal lymphoma simulates other neoplastic osseous surface tumors more than that of lymphoma in other locations. Consideration of this rare presentation of non-Hodgkin's lymphoma in the differential diagnosis of periosteal bone lesions can be helpful to ensure proper diagnosis and treatment. PMID- 12652340 TI - Rosai-Dorfman disease manifesting as a solitary lesion of the radius in a 41-year old woman. AB - Rosai-Dorfman disease is a rare entity predominantly affecting children and young adults, characterized in 83-95% of cases by painless bilateral cervical lymphadenopathy. We report the unusual case of a 41-year-old woman with Rosai Dorfman disease that presented as a solitary lesion of the radius without other clinical manifestations. PMID- 12652341 TI - Metastatic breast carcinoma to bone disguised by osteopoikilosis. AB - A case of metastatic lobular carcinoma of the breast in conjunction with osteopoikilosis is described. Widespread diffuse sclerotic bone lesions were identified on radiographs in a patient with breast carcinoma. In addition computed tomography demonstrated discrete spherical areas of increased density throughout the skeleton manifest typically by osteopoikilosis. No systemic symptoms were evident, blood parameters were normal and the lesions did not demonstrate any increased uptake of technetium on bone scan. An iliac crest bone biopsy, however, revealed metastatic disease in addition to osteopoikilosis. Conventional radiological investigations may not delineate metastasis on a background of bone dysplasia. PMID- 12652342 TI - Evidence-based medicine: applications in pediatric cardiology? PMID- 12652343 TI - Enhanced P2Y1 receptor expression in the brain after sensitisation with d amphetamine. AB - RATIONALE AND OBJECTIVES: Many pathological and physiological processes are associated with the transcriptional induction of specific receptors. The aim of the present study was to examine whether the development of d-amphetamine (AMPH) induced sensitisation is related to an altered P2Y(1) receptor expression. METHODS: Rats, treated for 5 successive days with AMPH (1.5 mg/kg, i.p.), alone or after pre-treatment with the non-specific P2 receptor antagonist pyridoxal phosphate-6-azophenyl-2,4-disulphonic acid (PPADS, 0.6 nmol, i.c.v.) and tested in an open field system with respect to locomotor response, were studied immunocytochemically 5 days after the last AMPH injection. RESULTS: In the behaviourally sensitised animals, astrogliosis, characterised by hypertrophy, increase in glial fibrillary acidic protein (GFAP) immunoreactivity (IR) and astrocytic proliferation in striatal areas and the nucleus accumbens were observed. Quantification of the P2Y(1) receptor stained cells revealed an increase in the receptor expression after AMPH-induced sensitisation in the studied regions. Pre-treatment with PPADS prior to each AMPH administration prevented the development of sensitisation, astrogliosis and P2Y(1) receptor up regulation. PPADS failed to alter the number of P2Y(1) receptor-labelled cells when given alone. Confocal laser scanning microscopy indicated the localisation of P2Y(1) receptors on GFAP-labelled astrocytes as well as on tubulin (betaIII) labelled neurones, under control conditions and after AMPH administration. CONCLUSION: The present results confirm the existence of P2Y(1) receptors on astrocytes and neurones as possible targets of endogenous ATP and in addition show their up-regulation as a consequence of P2Y(1) receptor-involvement in AMPH induced sensitisation in vivo. PMID- 12652344 TI - Role of the hypothalamic-pituitary-adrenal axis in reinstatement of cocaine seeking behavior in squirrel monkeys. AB - RATIONALE: Stress has been suggested to play a role in relapse to cocaine-seeking behavior. An important physiological system activated by stress is the hypothalamic-pituitary-adrenal (HPA) axis; however, evidence for a role of HPA axis activation in cocaine relapse has been contradictory. OBJECTIVES: This study examined the effects of pharmacological stimulation of the HPA axis on reinstatement of drug-seeking behavior and salivary cortisol levels in a non human primate model of cocaine relapse. In addition, the effect of corticotropin releasing hormone type 1 receptor (CRH-R1) blockade on cocaine priming-induced reinstatement was investigated. METHODS: Squirrel monkeys were trained to self administer cocaine under a second-order schedule in which behavior was maintained by IV drug injections and a drug-paired visual stimulus. A period of extinction was then imposed during which saline was substituted for cocaine and the stimulus was omitted. Subsequently, monkeys were tested for reinstatement of cocaine seeking following priming injections of drugs. During reinstatement tests, the drug-paired stimulus was restored. Salivary cortisol levels were determined to measure the effects of drug treatments on the HPA axis activity. RESULTS: Priming with corticotropin releasing hormone (10 and 50 microg/kg), adrenocorticotropic hormone (1 microg/kg), or cortisol (1-10 mg/kg) did not induce significant reinstatement of cocaine seeking. All of these treatments, however, resulted in a significant increase in salivary cortisol. In contrast, priming injections of cocaine (0.1-1.0 mg/kg) dose-dependently induced reinstatement of drug seeking, but did not increase salivary cortisol. The CRH-R1 antagonist CP-154,526 (10 mg/kg, IV) did not modulate cocaine priming-induced reinstatement of drug seeking, but attenuated CRH-induced increases in salivary cortisol. CONCLUSIONS: The results suggest that activation of the HPA axis is neither necessary nor sufficient for reinstatement of cocaine-seeking behavior in this non-human primate model of cocaine relapse. PMID- 12652345 TI - Behavioral, neuroendocrine and neurochemical effects of the imidazoline I2 receptor selective ligand BU224 in naive rats and rats exposed to the stress of the forced swim test. AB - RATIONALE: There is evidence for alterations in imidazoline(2) (I(2)) receptor density in depressed patients. Selective I(2) receptor ligands modulate central monoamine levels and activate the hypothalamo-pituitary-adrenal (HPA) axis and may have potential as antidepressants. OBJECTIVES: To study the behavioral effects of the selective I(2) receptor ligand BU224 in the rat forced swim test (FST) and its effects on the HPA axis and central monoaminergic responses. METHODS: Rats received saline or BU224 (10 mg/kg IP) 24, 18 and 1 h prior to 15 min exposure to the FST. Saline- and BU224-treated non-stressed groups were included. Time spent immobile, struggling and swimming calmly was measured. Plasma adrenocorticotrophic hormone (ACTH) and corticosterone levels 90 min post BU224 were measured in addition to tissue levels of monoamines and metabolites in the frontal cortex, hippocampus and hypothalamus. RESULTS: Administration of BU224 significantly reduced immobility and increased mild swimming without affecting struggling. Exposure to the FST significantly increased plasma ACTH and corticosterone levels. BU224 administration also increased ACTH and potentiated the ACTH response to FST with no effect on corticosterone. BU224 administration significantly increased frontal cortex 5-hydroxytryptamine (5-HT) levels and decreased 5-HT turnover in the frontal cortex and hypothalamus of rats exposed to FST. In non-stressed rats, BU224 decreased 5-HT turnover in the hippocampus and hypothalamus and decreased norepinephrine turnover in the frontal cortex. CONCLUSIONS: The selective I(2) receptor ligand BU224 reduces immobility of rats in the FST, indicative of antidepressant-like activity. This effect is accompanied by alterations in HPA axis and central monoaminergic activity. PMID- 12652346 TI - Brain circuitry and the reinstatement of cocaine-seeking behavior. AB - RATIONALE: Recent studies have attempted to identify the neuroanatomical substrates underlying primed reinstatement of drug-seeking behavior. Identification of neuronal substrates will provide a logical rationale for designing pharmacological interventions in treating drug relapse. OBJECTIVE: The objective was to identify brain circuitry that is shared between cue-, drug- and stress-primed reinstatement, as well as identifying aspects of brain circuitry that are distinct for each stimulus modality. The resulting circuit offers theoretical interpretations for consideration in future studies. RESULTS: Aspects of the circuitry mediating reinstatement can be identified with reasonable confidence. The role of the basolateral amygdala in cue-primed reinstatement, the role of the ventral tegmental area in drug-primed reinstatement and the role of adrenergic innervation of the extended amygdala in stress-primed reinstatement are well characterized. Also, all three modes for priming reinstatement may converge on the anterior cingulate cortex and have a final common output through the core of the nucleus accumbens. Lacunae in our understanding of the circuit were identified, especially with regard to how stress priming is conveyed from the extended amygdala to the shared anterior cingulate accumbens core circuit. CONCLUSIONS: The proposed convergence of priming stimuli into the glutamatergic projection from anterior cingulate to the accumbens core combined with the changes in glutamate transmission and signaling that accompany repeated psychostimulant administration points to the potential value of pharmacological agents that manipulate glutamate release or postsynaptic glutamate receptor signaling and trafficking in treating primed relapse in addicts. PMID- 12652347 TI - Clorgyline-induced switch from locomotion to mouthing in sensitization to the dopamine D2/D3 agonist quinpirole in rats: role of sigma and imidazoline I2 receptors. AB - RATIONALE: The monoamine oxidase inhibitor (MAOI) clorgyline, blocks locomotor sensitization to the D(2)/D(3) dopamine agonist quinpirole and sensitizes self directed mouthing behavior in rats by a mechanism independent of MAO inhibition. Clorgyline has a high affinity for imidazoline I(2) and sigma receptors, which could account for its effects on quinpirole sensitization. OBJECTIVES: To examine whether the effect of clorgyline on quinpirole sensitization is attributed to stimulation of either I(2) or sigma receptors. METHODS: In one experiment, rats received injections of the I(2) receptor agonist 2-BFI (0.2 mg/kg, IP) or vehicle, 90 min prior to each injection of quinpirole (0.5 mg/kg, SC, x 8, twice weekly) or saline. A similar protocol was used to examine the effects of the MAOI Ro 41-1049 (10 mg/kg, SC) on quinpirole sensitization. Unlike clorgyline, Ro 41 1049 has no affinity for sigma or I(2) sites. An initial experiment demonstrated that intermittent injections of clorgyline (1 mg/kg, SC) are as effective as a continuous clorgyline administration (1 mg/kg per day via osmotic mini-pump) on quinpirole sensitization. RESULTS: Like clorgyline, Ro 41-1049, but not 2-BFI, blocked the development of quinpirole-induced locomotor sensitization and induced instead sensitization of self-directed mouthing. CONCLUSIONS: Because Ro 41-1049 produced the same effects as clorgyline, and 2-BFI had no effects on quinpirole sensitization, it is unlikely that clorgyline exerts its effects via an action at sigma or I(2) receptors. Our results are consistent with the suggestion that clorgyline and Ro 41-1049 affect the behavioral response to quinpirole via the MAOI-displaceable quinpirole binding (MQB) site, and the hypothesis that the MQB site selects what motor output becomes sensitized to repeated injections of quinpirole. PMID- 12652348 TI - Effects of chronic d-amphetamine treatment on cocaine- and food-maintained responding under a progressive-ratio schedule in rhesus monkeys. AB - RATIONALE: d-Amphetamine is a candidate agonist medication for the treatment of cocaine dependence, and evaluation of d-amphetamine effects on abuse-related effects of cocaine in preclinical assays is warranted. OBJECTIVE: This study was designed to assess the effects of chronic d-amphetamine treatment on cocaine- and food-maintained responding under a progressive-ratio schedule in rhesus monkeys. The effects of schedule manipulations on cocaine and food-maintained responding were also examined for comparison with d-amphetamine effects. METHODS: Key-press responding under a progressive-ratio schedule resulted in the delivery of cocaine (0.032 mg/kg per injection) or 1 g food pellets. The effect of manipulating cocaine dose (saline, 0.001-0.1 mg/kg per injection) or the number of food pellets delivered (0, 1 and 4 pellets) was determined. Subsequently, three schedule parameters were manipulated: (1) starting ratio value, (2) increments of the ratio progression, and (3) duration of post-reinforcer time-outs when the ratio value was constant. Finally, the effects of 10-day treatment with d amphetamine (0.01-0.1 mg/kg per hour) were examined. RESULTS: Break points increased as a function of cocaine dose or the number of food pellets, and similar break points were maintained by delivery of 0.032 mg/kg per injection cocaine and 1 food pellet. Manipulation of schedule parameters produced similar effects on responding maintained by cocaine (0.032 mg/kg per injection) or food (1 pellet). In contrast, d-amphetamine produced a dose-dependent decrease in cocaine-maintained responding and had less consistent effects on food-maintained responding. CONCLUSIONS: These results are consistent with the hypothesis that chronic treatment with d-amphetamine decreases cocaine self-administration in rhesus monkeys, possibly by attenuating the reinforcing effects of cocaine. PMID- 12652349 TI - Essential conservation of D1 mutant phenotype at the level of individual topographies of behaviour in mice lacking both D1 and D3 dopamine receptors. AB - RATIONALE: In the absence of agonists and antagonists evidencing appropriate selectivities, individual and interactive properties of D(1) and D(3) dopamine receptors would be illuminated most powerfully by their co-deletion. OBJECTIVES: To define and contrast the behavioural phenotype of D(1)/D(3) double knockout mice in comparison with wild types, and with individual D(1) and D(3 )mutants. METHODS: Behavioural phenotype was characterised using an ethologically based topographical technique. RESULTS: On comparison with wild types, D(1)/D(3) double mutants were characterised topographically as follows: increases in sniffing and locomotion, which evidenced delayed habituation; reductions in rearing free, rearing seated, grooming, chewing and stillness. Though the D(1)/D(3) double mutant ethogram comprised elements of both single mutant D(1) and D(3) lines, this phenotype was largely reflective of the D(1) mutant component. CONCLUSIONS: Distinct patterns of initial exploratory behaviour and of temporal change over subsequent habituation were evident across the three genotypes, with particular conservation of the D(1) phenotype in D(1)/D(3 )double mutants. Under the present conditions, there was little systematic evidence for D(1):D(3) interactions in the regulation of these aspects of behaviour. PMID- 12652351 TI - Foreword. PMID- 12652352 TI - Loss of early insulin secretion leads to postprandial hyperglycaemia. AB - AIMS/HYPOTHESIS: A loss of early-phase insulin response is common in Type II diabetic patients and in people with impaired glucose metabolism. It is hypothesized that this alteration is not simply a marker for the risk of developing diabetes, rather it is a an important pathogenetic mechanism causing excessive postprandial hyperglycaemia. METHODS: Relevant literature on the epidemiology, physiopathology, and therapeutic intervention related to loss of early insulin secretion and postprandial hyperglycaemia has been analysed. RESULTS: In response to intravenous glucose, insulin secretion is biphasic. This behaviour translates as a rapid release of insulin into the blood stream in response to the ingestion of carbohydrates or a mixed meal. The rapid increase in portal insulin concentration and the avid binding of the hormone to its receptor on liver cell membranes, account for a prompt suppression of endogenous glucose production and reduction of the rate of increase in plasma glucose concentrations. This has been supported by experimental studies carried out in both animals and humans: the selective abolition of early insulin secretion in healthy subjects results in impaired glucose tolerance, excessive glucose excursions, and possible hampering of the thermic effects of ingested carbohydrates. In non-diabetic subjects, the loss of early insulin secretion is a determinant for developing diabetes. The critical role of the early-phase insulin response in determining postprandial hyperglycaemia, is supported by an amelioration of glucose tolerance by restoring the acute rise in plasma insulin concentrations after the ingestion of both glucose and a mixed meal. This amelioration in plasma glucose profile can prevent late hyperglycaemia and hyperinsulinaemia. CONCLUSION/INTERPRETATION: Therapeutic approaches aimed at restoring a physiological pattern of insulin secretion could prove effective in reducing postprandial glucose excursions particularly in the early stage of Type II diabetes. PMID- 12652350 TI - Procalcitonin and C-reactive protein plasma concentrations in nonseptic uremic patients undergoing hemodialysis. AB - OBJECTIVE: To assess procalcitonin (PCT) and C-reactive protein (CRP) plasma concentrations and clearance in nonseptic end-stage renal failure patients undergoing their first three hemodialysis sessions. DESIGN AND SETTING: Prospective observational consecutive clinical study at a university hospital. PATIENTS: The study recruited 55 end-stage renal failure patients without evidence of systemic infection undergoing the creation of an arteriovenous fistula to start hemodialysis for the first time. INTERVENTIONS: Blood samples were collected before and after each of the first three (4-5 h) hemodialysis sessions. PCT was assayed by immunoluminometry. MEASUREMENTS AND RESULTS: The mean plasma concentration of PCT prior to the first three hemodialysis sessions declined significantly following each session. There was no significant difference between CRP plasma concentrations before and after hemodialysis sessions. CONCLUSIONS: The presence of an elevated PCT in plasma of not yet dialyzed uremic nonseptic patients indicates that uremia per se and not the dialysis process is the origin of such elevation. PCT levels declined with successive hemodialysis sessions. We propose that in the not yet dialyzed uremic nonseptic patients a baseline PCT level of approx. 1.5 ng/ml should be expected. Although the mean plasma CRP level was elevated, hemodialysis had no significant effect on CRP concentration, making CRP a possible useful marker of sepsis in these patients. PMID- 12652353 TI - The possible role of postprandial hyperglycaemia in the pathogenesis of diabetic complications. AB - It is both common and wise practice to adjust the treatment of diabetic patients to obtain plasma glucose concentrations as close as possible to the "normal range", correcting both postprandial hyperglycaemic spikes and the less increased, but persistently high, plasma glucose concentration between meals. The Diabetes Control and Complications Trials (DCCT) and the United Kingdom Prospective Diabetes Study (UKPDS) have provided evidence that intensive treatment can prevent complications associated with diabetes mellitus. In both studies, effectiveness of hyperglycaemic treatment was assessed by means of the glycated haemoglobin level. This is an integrated measure of both postprandial and fasting hyperglycaemia. The absolute and relative importance of the two conditions, however, could differ depending upon the organ or system suffering from diabetic complications and other more or less known individual factors. This paper aims to emphasize the effects of acute hyperglycaemia, in particular, postprandial hyperglycaemia, on the development of diabetic complications. The role of oxidative stress as a mediator of acute hyperglycaemia is further discussed. More investigation in this area is required so that treatment can be eventually individualised. Perhaps, in some patients, efforts could be concentrated on the control of hyperglycaemic spikes and/or specific organ or system susceptibility to either acute or chronic hyperglycaemia. PMID- 12652354 TI - Post-challenge hyperglycaemia is associated with premature death and macrovascular complications. AB - Unravelling the precise association between cardiovascular disease and Type II (non-insulin-dependent) diabetes mellitus has proved problematic, largely as a result of the confusing variety of diagnostic criteria and methodologies used in previous studies of disease-associated risk. More recently, improved standardization and continuing refinements to risk analyses have begun to clarify the relative importance of individual risk factors for macrovascular complications and progression to cardiovascular disease. The evidence accumulated from several large-scale epidemiological and intervention studies strongly indicates that hyperglycaemia and specifically the post-load hyperglycaemia resulting from postprandial glucose excursions is the main factor associated with increasing the risks of morbidity and mortality in Type II diabetes. The DECODE study has de-emphasized the importance of fasting glucose, in contrast to post load hyperglycaemia, as the principal measurable determinant of exposure to the risk of cardiovascular disease in Type II diabetes. Similarly, the Diabetes Intervention Study identified mealtime hyperglycaemia as an independent risk factor for myocardial infarction and all-cause mortality, while several other studies have also shown that such mortality is better predicted from post challenge measurement criteria rather than from fasting-glucose criteria. It is likely that postprandial hyperglycaemia is an independent risk factor for atherosclerosis and is also exerting an atherogenic effect indirectly through the clustering of a number of risk factors for atherosclerosis. Whether the relation between post-load hyperglycaemia and the risk of cardiovascular events is causal is still not known. PMID- 12652355 TI - Pharmacologic restoration of the early insulin response in pre-diabetic monkeys controls mealtime glucose excursions without peripheral hyperinsulinaemia. AB - AIMS/HYPOTHESIS: This study sought first to compare the pharmacodynamics and pharmocokinetics of two rapid-onset, rapidly-reversible insulinotropic agents, nateglinide and repaglinide, in pre-diabetic Cynomolgus monkeys and second to use these agents to assess the metabolic effects of early insulin secretion on prandial glucose control. METHODS: First, equipotent doses of nateglinide (20 mg/kg) and repaglinide (0.1 mg/kg) or vehicle were given intragastrically to overnight-fasted ketamine-anesthetized pre-diabetic Cynomolgus monkeys and samples were obtained for measurement of plasma glucose, insulin, glucagon, NEFA and drug concentrations. Second, nateglinide, repaglinide or vehicle were administered 10 min before a glucose-supplemented liquid meal and prandial glucose and insulin profiles were compared. RESULTS: Although oral administration of nateglinide and repaglinide elicited similar maximum increments of plasma insulin (+403 and +448 pmol/l, respectively), the effects of nateglinide were more rapidly manifest and less prolonged. With nateglinide, insulin increased within 10 min and returned to baseline within 50 min. After repaglinide, the first increase occurred at 30 min and insulin concentrations remained increased for 3.5 h post-dose. When given 10 min before a meal, nateglinide increased early, but not total insulin release (AUC(0-210)=108 vs 150 nmol/l min for nateglinide and vehicle, respectively) and reduced prandial glucose excursions by 78%. Repaglinide increased total insulin release (AUC(0-210)=298 nmol/l min) and reduced glucose excursions by 53%. CONCLUSION/INTERPRETATION: Nateglinide is more rapid-acting and rapidly-reversible than is repaglinide. By restoring a more physiologic insulin profile, nateglinide is more effective than repaglinide in controlling prandial glucose excursions with less hyperinsulinaemia. PMID- 12652356 TI - The role of oral antidiabetic agents: why and when to use an early-phase insulin secretion agent in Type II diabetes mellitus. AB - Evidence obtained in the 1990's strongly supports the notion that glycaemic control is important not only in Type I (insulin-dependent), but also in Type II (non-insulin-dependent) diabetes mellitus. Although measurement of HbA(1c) is the standard for assessing the effect of glucose control in the occurrence and prevention of diabetic sequelae, more recent evidence indicates that other glucose parameters are also important. Postchallenge and postprandial hyperglycaemic peaks seem to be prospective determinants of vascular damage in early Type II diabetes. Currently, there is no overall accepted standard approach for the pharmacological management of Type II diabetes. The United Kingdom Prospective Diabetes Study has shown that reaching a near-normal glycaemic target is critically important and the pharmacotherapy of this progressive disease is difficult. Loss of endogenous insulin secretion has been substantiated to cause the progression of Type II diabetes in the United Kingdom Prospective Diabetes Study. Early insulinization, however, was not advantageous over other forms of therapy. The advent of polypharmacy in recent years has greatly strengthened the treatment of this disease. This synergy has been extended of late with the development of early-phase insulin secretion agents. Two such agents, nateglinide and repaglinide, can be used to reduce mealtime glucose excursions and HbA(1c) as monotherapy, and in combination with metformin; their antidiabetic potential is similar to the combination treatment with glibenclamide and metformin. Additional substantiation of their long-term effect on improving life expectancy and reducing diabetic complications in Type II diabetic patients is now required. PMID- 12652357 TI - The mechanisms underlying the unique pharmacodynamics of nateglinide. AB - Nateglinide, a D-phenylalanine derivative, belongs to a new group of insulinotropic agents with rapid onset and short duration of action. These agents have been developed to reduce the risk of hypoglycaemia associated with pharmacological control and to decrease the likelihood of pancreatic beta-cell exhaustion. Nateglinide mediates the release of insulin from beta-cells by binding to the sulphonylurea receptors, which leads to the closure of ATP sensitive K(+) channels. Increasing evidence from receptor binding, mechanistic and in vitro and in vivo insulin studies indicate unique pharmacodynamic and pharmacokinetic properties with nateglinide that are distinct from those of sulphonylureas. The time required by nateglinide to close beta-cell K(ATP) channels is comparable to that of glyburide but threefold and fivefold faster than repaglinide and glimepiride, respectively. Furthermore, its effects are rapidly reversed with an off-rate at the K(ATP) channel twice as fast as that of glyburide and glimepiride and five times faster than repaglinide. This results in a rapid and short insulin response characteristic of the physiological pattern of post-mealtime insulin release. Internalisation into beta-cells is not required for the action of nateglinide. Given that the kinetic profile of the agent is associated with selective enhancement of early-phase insulin secretion, nateglinide is expected to minimise post-meal hyperglycaemia with minimal propensity for hypoglycaemia. PMID- 12652358 TI - Rationale and options for combination therapy in the treatment of Type 2 diabetes. AB - Pharmacological therapy for Type 2 (non-insulin-dependent) diabetes mellitus aims at controlling hyperglycaemia to delay or prevent complications associated with the disease. Most patients with Type 2 diabetes present with both stimulated insulin deficiency and insulin resistance. In general, the former can manifest as postprandial hyperglycaemia and the latter as fasting hyperglycaemia, though a definitive association has not been established. Emerging data show a high failure rate of long-term monotherapy and establishes the significance of mealtime glycaemia and the role of postprandial glucose excursions in the development and progression of vascular complications. To overcome such failures of monotherapy and to address the different underlying defects of the pathology of Type 2 diabetes, a combined therapy of oral antidiabetic agents with complementary modes of action should be considered. Currently used oral antidiabetic agents such as sulphonylureas, biguanides (metformin) and the thiazolidinediones (rosiglitazone, pioglitazone) commonly target fasting hyperglycaemia and have limited additive effects on postprandial glycaemia. In contrast, alpha-glucosidase inhibitors can reduce postprandial hyperglycaemia but gastrointestinal side effects restrict their use. The development of new agents to control postprandial glucose excursions could be considered as an additional objective for the management of Type 2 diabetes. To this end new short-acting enhancers of insulin secretion such as repaglinide (benzoic acid derivative) and nateglinide (amino acid derivative) have been developed. The combination of such agents with other complementary modes of action, e.g. an insulin sensitizer, could target better major underlying defects of Type 2 diabetes and thereby provide a better approach for controlling the entire glycaemic risk. PMID- 12652359 TI - The middle cranial fossa approach: an anatomical study. AB - Hearing preservation surgery has become an option for an increasing number of patients with vestibular schwannomas due to diagnosis at an earlier stage. The middle cranial fossa approach represents one such surgical approach for resection of vestibular schwannomas with hearing preservation. We have undertaken an anatomical study of the middle cranial fossa approach to the internal auditory meatus using 20 fresh temporal bones. By simulating the surgical approach it was possible to analyze critically two of the main recognized subapproaches to the internal acoustic meatus. The results confirmed that the angle subtended by the facial nerve and "blue-lined" semicircular canal was much less than 60 degrees but equally important was the degree of individual variability. Furthermore the roof of the geniculate fossa was not infrequently dehiscent. The distance measured from the inner table of the craniotomy to the superior semicircular canal was on average 22 mm, similar to previous reports and utilized by some in their approach in this challenging surgery. From this anatomical study it appears that safe dissection of this area is facilitated by observing the more acute angle between the facial nerve and superior semicircular canal and by taking advantage of the relationship between the inner table and important landmarks. PMID- 12652361 TI - Preliminary results and evidence of early regeneration in inferior alveolar nerve fibers. AB - Inferior alveolar nerve (IAN) damage can occur in trauma, cyst enucleation, sagittal split osteotomy or third molar removal, and the consequences are a loss of sensation to the mandibular teeth, gingiva and lower lip. Because of its anatomical position in a bony canal, IAN suture is rarely evoked. The aim of this study was to demonstrate the reality of IAN regeneration by using electrophysiological and histological methods after experimental section and suture of this nerve in rabbits. Nine adult female animals were used for the experiments. Six months after section and suturing using 10.0 nylon with a conventional technique, electrical stimulation of the nerve was performed to record electrophysiological activity. Each rabbit was its own reference. In each case, an action potential was recorded after microsurgical repair and definitively suppressed by section of the nerve. Morphometric analysis showed a decrease in the number of nerve fibers in the operated nerve versus the control nerve. The histological study showed an increase in nerve fibers with a cross sectional area of 19-36 and 37-73 micro m(2) and a decrease in the smaller fibers (2-4 and 5-7 micro m(2)). This preliminary study confirms the possibility of nerve regeneration in rabbits 6 months after section and conventional suturing. PMID- 12652362 TI - Morphometric measurements from various reference points in the orbit of male Caucasians. AB - The aim of this study was to determine the morphometric variations from various reference points to decrease risks in orbital surgery. Sixty-two orbits obtained from 31 skulls of male adult Caucasians were measured with a millimetric compass. On the medial orbital wall, the midpoint of the anterior lacrimal crest was the reference point; from this point we measured distances of 23.9+/-3.3 mm, 35.6+/ 2.3 mm, 41.7+/-3.1 mm and 6.9+/-1.5 mm respectively to the anterior ethmoidal foramen, posterior ethmoidal foramen, midpoint of the medial aspect of the optic canal and posterior lacrimal crest. On the same wall, distances from the plane of the anterior and posterior ethmoidal foramina to the ethmoido-maxillary suture and distance from the posterior ethmoidal foramen to the anterior ethmoidal foramen and midpoint of the medial margin of the optic canal were 14.9+/-2.3 mm, 9.8+/-2.9 mm and 6.8+/-2.2 mm respectively. On the inferior orbital wall, the main reference point was the infraorbital foramen, and from this point to the midpoints of the lateral margin of the fossa for the lacrimal gland, inferior orbital fissure, inferior orbital rim and inferior aspect of the optic canal was 23.8+/-7.2 mm, 31.9+/-3.9 mm, 6.7+/-1.9 mm and 50.3+/-3.2 mm respectively. On the superior orbital wall, the distances from the supraorbital foramen to the midpoints of the superior orbital fissure, fossa for the lacrimal gland and superior aspect of the optic canal were 45.7+/-3.6 mm, 26.0+/-2.5 mm and 45.3+/ 3.2 mm respectively. Furthermore, on the same wall, the distance from the posterior ethmoidal foramen to the midpoint of the superior orbital fissure was 14.6+/-2.8 mm. Finally, on the lateral orbital wall the frontozygomatic suture was the reference point. From this point distances to the midpoints of the fossa for the lacrimal gland, superior orbital fissure, lateral aspect of the optic canal and inferior orbital fissure were 17.5+/-2.1 mm, 37.7+/-3.6 mm, 44.9+/-2.5 mm and 33.4+/-3.1 mm respectively. PMID- 12652363 TI - McBurney's point: are we missing it? AB - A prospective study of 100 post-evacuation barium enemas was done. Films were centered at McBurney's point, with an opaque skin marker at that point. Analysis of these revealed that in only one case (1%) was the base of the appendix at McBurney's point. In 67% it was cephalic and in 32% it was caudal to this point. The limitations of McBurney's point as an anatomical landmark should be recognized. This needs to be highlighted in teaching anatomy, especially to surgical trainees. Planning and choice of surgical incisions should be based on an understanding of these anatomical variations since McBurney's original description was clinical rather than anatomical. PMID- 12652364 TI - The venous drainage of the lunate bone. AB - One of the theories as to the etiology and pathogenesis of lunatomalacia (Kienbock's disease) is based on the presentation of an osseous compartment syndrome caused by a venous block in the pedicle. This gave us cause to examine the lunate bone more closely and to investigate possible anatomical causes for the disruption. For this purpose, ten hands were removed from cadavers proximal to the radiocarpal articular space. Through an artificial intraosseous canal, which did not touch the vascular structures of the lunate bone, epoxy could be injected under controlled conditions. The venous drainage, from the exit out of the bones up to the entrance into the comitant veins of the distal forearm, was exposed as a preparation under the microscope. In all preparations a dense plexus of small venous vessels was found at the palmar and dorsal periosteal face which has not previously been described in literature. As this wide plexus is woven into the solid palmar as well as into the dorsal connective tissue, it could be, as we suppose, the weak point of the venous drainage we have been looking for. It is easy to imagine that the rheological situation in this venous segment is impaired and an osseous compartment syndrome is induced by systemic factors as well as by local compression. PMID- 12652365 TI - The seated man: geometry acquisition and three-dimensional reconstruction. AB - A three-dimensional (3D) virtual human body in seated position can be used in a wide range of applications such as basic or clinical research. A male cadaver placed in the driving position in a car cockpit was frozen. Strictly parallel 5 mm thickness serial sections were then performed. Both sides of the slices were photographed, scanned and analyzed by anatomists with software tools, in order to produce accurate contours of the identified anatomical structures. A 3D reconstruction of organs, based on the marching cubes method, was achieved except for thin structures. This allowed an anatomical validation of these reconstructed elements. The result of this work is a complete geometrical database of a seated human body in the driving position including skin, all bones, internal organs and main muscle groups. PMID- 12652366 TI - Three-dimensional imaging of the petrous bone for the middle fossa approach to the internal acoustic meatus: an experimental study. AB - The goal of this study was to highlight the feasibility of creating three dimensional (3D) pictures of the petrous bone from a routine CT examination which can be used for a middle fossa approach to the internal acoustic meatus, in order to secure this operation. The surgical aim is to reach the roof of the internal acoustic meatus directly without injuring the adjacent functional structures of the petrous bone. Two heads of embalmed cadavers were scanned every millimeter with a slice thickness of 1 mm centered on the petrous bones. The horizontal reference was the Frankfurt line and the frontal and sagittal planes were perpendicular to this line. This method is similar to routine examinations for surgical patients. The pictures were first loaded on an optical disk, then into a computer (Silicon Graphics System). Amira software was used to create 3D pictures. The anatomy of the temporal bone could easily be identified, notably the surgical landmarks of the middle fossa approach. Three-dimensional computer assisted imaging can reveal the anatomy of the petrous bone in a realistic view. The main anatomic structures for a middle fossa approach can be recognized easily. This realistic view may be very useful for surgeons, and 3D images deserve to be developed further. PMID- 12652367 TI - Personal-computer-based three-dimensional reconstruction and simulation of maxillary sinus. AB - Anatomical descriptions of the maxillary sinus are critical in pathological diagnosis and the treatment planning of surgical procedures. This study was undertaken to develop a new technique for simulating anatomical structures and to clarify the morphological and clinical characteristics of the maxillary sinus. Thirty-three hemi-sectioned Korean heads were used in this study. CT scans and DentaScan reformatted cross-sectional images were taken on all specimens. From the CT images, three-dimensional reconstructed images were made using the V-works program. From the three-dimensional reconstructed images of the maxillary sinus, six categories of maxillary sinus were created, categorized according to their lateral aspects and shapes of the inferior walls. In 55%, a flat inferior wall of the maxillary sinus was observed. All measurements (anterior-posterior length, height, width and volume) of the sinus were larger in males than in females. From the DentaScan reformatted panoramic images, the anterior limit of the maxillary sinus was located in the first premolar area (58%), and the posterior limit was in the third molar and maxillary tuberosity area (94%). We therefore offer a new virtual technique for manipulating three-dimensional reconstructed images easily on a personal computer. On the reconstructed images the three-dimensional morphology could be observed and the anatomical characteristics of the maxillary sinus and surrounding structures could be determined. PMID- 12652368 TI - Anatomic variations of the paranasal sinus area in pediatric patients with chronic sinusitis. AB - Endoscopic endonasal surgery has been performed in children. Therefore, we need to know the precise anatomy and anatomic variations of the lateral nasal wall. This is important in order to achieve better surgical results and avoid complications. We also need to know the relationship between the anatomic variations and sinonasal disease. For the purpose of this study we assessed the anatomic variations of the lateral nasal wall and the association of these variations with chronic sinusitis in children. Forty-seven children with chronic sinus disease were included in the study. There were 25 female and 22 male patients with ages ranging from 2 to 16 years (mean 10.5+/-3.8 years). On coronal and axial computed tomographic (CT) scans, the anatomic variations of the lateral nasal wall and sinusitis were assessed. A pneumatized middle concha (MC) was the most common anatomic variation, followed by pneumatization of the superior concha (SC), Haller cell and agger nasi cell. Secondary MC, large ethmoidal bulla, maxillary sinus hypoplasia, Onodi cell, and uncinate process pneumatization were relatively rare. Maxillary sinusitis was the most common sinus infection, followed by ethmoidal, sphenoidal and frontal sinusitis in that order. There was no significant relationship between the sinusitis and anatomic variations except for some minor associations. In conclusion, anatomic variations in the lateral nasal wall are common in children. Local, systemic or environmental factors appear to be more important in pediatric sinusitis than the anatomic variations. Due to the absence of a definitive relationship between the anatomic variations and sinus disease, aggressive surgical interventions should be avoided while performing endonasal endoscopic surgery in the children. PMID- 12652369 TI - A rare accessory coracobrachialis muscle: a review of the literature. AB - During previous dissections to investigate the incidence of the third head of the biceps brachii in neonate cadavers an accessory coracobrachialis muscle was unilaterally found in a neonate male cadaver. This muscle originated from the coracoid process and the capsule of the shoulder joint. It was inserted into the antebrachial fascia and the medial epicondyle of the humerus. The coracobrachialis muscle is used as a transposition flap in deformities of infraclavicular and axillary areas and in postmastectomy reconstruction. It is also a guide to the axillary artery during surgery and anesthesia. This additional muscle may cause musculocutaneous or high median nerve paralysis. We were unable to find any similar case in the published literature. The anatomic variations of the coracobrachialis muscle may cause confusion during surgery or evaluation of CT and MRI scans. PMID- 12652371 TI - Introduction. Integrating nutrition therapy into medical management of human immunodeficiency virus. PMID- 12652372 TI - General nutrition management in patients infected with human immunodeficiency virus. AB - Nutritional management is integral to the care of all patients infected with human immunodeficiency virus (HIV). HIV infection results in complicated nutritional issues for patients, and there is growing evidence that nutritional interventions influence health outcomes in HIV-infected patients. We define levels of nutritional care, and we discuss when patients should be referred to providers (i.e., registered dietitians) with nutritional and HIV expertise. PMID- 12652373 TI - Assessment of nutritional status, body composition, and human immunodeficiency virus-associated morphologic changes. AB - Nutritional status should be assessed at regular intervals as part of management of human immunodeficiency virus (HIV) infection. The simplest approach to assessment is serial weight measurement. A comprehensive nutritional assessment includes (1) anthropometric measurements of body composition; (2) biochemical measurements of serum protein, micronutrients, and metabolic parameters; (3) clinical assessment of altered nutritional requirements and social or psychological issues that may preclude adequate intake; and (4) measurement of dietary intake. Techniques for measuring body composition of fat and lean body mass include anthropometry and bioelectric impedance analysis. Other techniques, including dual X-ray absorptiometry (DXA), hydrodensitometry, total body potassium measurement, and cross-sectional computed tomography or magnetic resonance imaging are available in research centers. Anthropometry, including waist-hip ratios, regional DXA, and cross-sectional imaging, is best for detecting morphologic changes associated with fat redistribution syndrome. Nutritional assessment and intervention in children with HIV can help to prevent stunted growth and development. PMID- 12652375 TI - Lipid abnormalities. AB - Dyslipidemia is an important clinical problem in individuals infected with human immunodeficiency virus (HIV) who are receiving antiretroviral therapy. Data suggest that increased cardiovascular disease is occurring in this population. HIV-infected individuals should undergo evaluation and treatment regimens based on the current National Cholesterol Education Program guidelines. In most situations, the first interventions should be nonpharmacological and should include diet, exercise, and management of other hygienic risk factors. If pharmacologic therapy becomes necessary, the choices of lipid-lowering agents should be limited to agents with the least likelihood of adverse drug interactions. PMID- 12652374 TI - Weight loss and wasting in patients infected with human immunodeficiency virus. AB - Weight loss and muscle wasting remain significant clinical problems, even in the era of potent antiretroviral therapy. In patients infected with human immunodeficiency virus (HIV), wasting, particularly loss of metabolically active lean tissue, has been associated with increased mortality, accelerated disease progression, loss of muscle protein mass, and impairment of strength and functional status. Factors that may contribute to wasting include inadequate intake, malabsorptive disorders, metabolic alterations, hypogonadism, and excessive cytokine production. Evidence now demonstrates that nutritional counseling and support, appetite stimulants, progressive resistance training, and anabolic hormones can reverse weight loss and increase lean body mass in HIV infected patients. Despite a growing body of evidence on the importance of nutritional intervention to prevent wasting in adults, maintain growth velocity in children, and promote restoration of weight and lean body mass in stable, low weight patients, no therapeutic guidelines currently exist for the management of weight loss and wasting in HIV-infected patients. Principles and guidelines for assessment and management of weight loss and wasting in patients with HIV/AIDS are presented. PMID- 12652376 TI - Body habitus changes related to lipodystrophy. AB - Changes in body fat in persons infected with the human immunodeficiency virus (HIV) have been associated with deleterious changes in blood lipids and insulin resistance, raising concern that these changes will increase the risk for accelerated atherosclerosis. Changes in body fat are often identified in advanced disease but may also occur early after HIV infection is detected. Conflicting evidence suggests that fat maldistribution may be related to use of protease inhibitors, nonnucleoside reverse transcriptase inhibitors, or a combination of these two classes of drugs, but the etiologies of the various changes in body fat remain uncertain. To date there have been no remedies for the loss of subcutaneous fat, but recent evidence has suggested that discontinuation of stavudine or zidovudine therapy may be associated with limited restoration of extremity fat. For fat accumulation, a number of strategies have been attempted, including treatment with human growth hormone, androgens, or metformin, and changes in diet and exercise. As in persons not infected with HIV, it is expected that the cornerstone of management, especially in the presence of central obesity, dyslipidemia, and insulin resistance, will include a diet low in saturated fat, with low-glycemic index carbohydrates, and high in fiber. Very limited evidence in persons infected with HIV has suggested that a supervised exercise program may be beneficial. PMID- 12652377 TI - Insulin and carbohydrate dysregulation. AB - Patients with human immunodeficiency virus receiving highly active antiretroviral therapy (HAART) may experience abnormal body composition changes as well as metabolic abnormalities, including dyslipidemia, increases in triglycerides, low high-density lipoprotein cholesterol levels, and abnormal carbohydrate metabolism, ranging from insulin resistance with and without glucose intolerance to frank diabetes. Whether the body composition changes (i.e., increased visceral adiposity and fat wasting in the peripheral tissues) are linked to abnormalities in carbohydrate metabolism is unclear. The use of HAART with and without therapy with protease inhibitors (PIs) is related to carbohydrate abnormalities and changes in body composition. Regimens that include PIs appear to have a higher incidence of insulin resistance (up to 90%) and diabetes mellitus (up to 40%). The etiology of these abnormalities is not well understood; what is known about insulin and carbohydrate dysregulation with HAART is discussed. PMID- 12652378 TI - Lactic acidemia in infection with human immunodeficiency virus. AB - Lactic acidosis in patients infected with the human immunodeficiency virus was initially identified as a rare complication of therapy with nucleoside analog reverse transcriptase inhibitors (NRTIs). The only patient group that appears to be at greater risk is pregnant women. More recently, milder elevations in lactate (i.e., lactic acidemia or hyperlactatemia) have been found to be more common and to be associated with numerous illnesses. Mild asymptomatic lactic acidemia is common, but it appears to lead to more severe illness only rarely. This suggests that routine measurement of plasma lactate should be limited to patients with previous acidemia who reinitiate NRTI therapy and to pregnant women. For symptomatic lactic acidemia (generally >5 mmol/L), NRTIs and other antiretroviral therapy should be ceased. Currently, asymptomatic lactic acidemia should not be treated and should not lead to a change in antiretroviral therapy. PMID- 12652379 TI - Emerging bone problems in patients infected with human immunodeficiency virus. AB - Recently, a high incidence of osteopenia and osteoporosis has been observed in individuals infected with human immunodeficiency virus (HIV). This problem appears to be more frequent in patients receiving potent antiretroviral therapy. Other bone-related complications in HIV-infected individuals, including avascular necrosis of the hip and compression fracture of the lumbar spine, have also been reported. People living with HIV have significant alterations in bone metabolism, regardless of whether they are receiving potent antiretroviral therapy. The underlying mechanisms to account for these observations remain unknown, although studies are underway to examine the relationship between the bone abnormalities and other complications associated with HIV and antiretroviral therapy. HIV infected patients with osteopenia or osteoporosis should be treated similarly to HIV-seronegative patients with appropriate use of nutritional supplements (calcium and vitamin D) and exercise. Hormone replacement and antiresorptive therapies might be also indicated. PMID- 12652380 TI - Food and water safety for persons infected with human immunodeficiency virus. AB - Public health and food safety experts estimate that millions of episodes of illnesses annually can be traced to contaminated food and water. Food and water safety is extremely important to persons infected with the human immunodeficiency virus (HIV) or with acquired immunodeficiency syndrome (AIDS). A compromised immune system causes people with HIV or AIDS to be more susceptible to foodborne illness from eating foods that are unsafely handled and poorly prepared and from using water from unsafe sources. Food- and waterborne illnesses can cause diarrhea, nausea, and vomiting that can lead to weight loss. These illnesses can be minimized or prevented if proper precautions are taken. PMID- 12652381 TI - Risk factors for primary bacteremia and endovascular infection in patients without acquired immunodeficiency syndrome who have nontyphoid salmonellosis. AB - This study sought to find the risk factors for primary bacteremia, endovascular infection, and in-hospital death for patients without acquired immunodeficiency syndrome who have nontyphoid salmonellosis. From September 1995 through September 2001, 301 patients with nontyphoid salmonellosis were admitted to our hospital; of these patients, 121 had primary bacteremia, and 28 had endovascular infection. Of the 121 patients with primary bacteremia, 64 were aged >50 years, and 26 had endovascular infection. Overall, 90 patients (29.9%) had immunodeficiency. Predictors of primary bacteremia were age; presence of systemic lupus erythematosus; group B, group C, or group D Salmonella infection; and immunodeficiency. The positive predictor of endovascular infection in adult patients with primary bacteremia was group C Salmonella infection, and negative predictors were immunodeficiency and solid-organ cancer. The overall in-hospital mortality rate was 12%; for primary bacteremia, it was 24.8%; for endovascular infection, it was 14.3%. Predictors of in-hospital death were age, extraintestinal infection, and solid-organ malignancy. PMID- 12652382 TI - Risk factors for endovascular infection due to nontyphoid salmonellae. PMID- 12652383 TI - Value of self-reportable screening criteria to identify asymptomatic individuals in the general population for urogential Chlamydia trachomatis infection screening. AB - Submission of samples from the home allows screening for Chlamydia trachomatis without preceding professional assessment of clinical risk factors. Therefore, a validation of self-reportable information for use as selective screening criteria is needed. We asked a total of 1175 women and 1033 men who participated in an in home sampling screening study to provide information on behavior and sociodemographic characteristics. In a multivariate model, selective screening criteria were developed on the basis of information from a random part of the tested population (development group), and the validity was assessed for the remaining part of the tested population (validation group). To find all infections, 95% of the subjects had to be screened, and screening 63% of them would have detected 86% of infections. Low predictive values were found when selective screening criteria from other studies were assessed. Selective screening by means of in-home sampling strategies among men and women aged 21-23 years cannot be recommended in the area studied. PMID- 12652384 TI - Outcome of penicillin-susceptible streptococcal prosthetic joint infection treated with debridement and retention of the prosthesis. AB - Debridement with retention of the prosthesis was the initial treatment modality for 19 cases of penicillin-susceptible streptococcal prosthetic joint infection that occurred in 18 patients who presented to the Mayo Clinic (Rochester, Minnesota) during 1969-1998. All of the cases of prosthetic joint infection occurred >30 days after implantation of the prosthesis, which was well fixed at the time of debridement. The median duration of symptoms before debridement was 4 days (range, 1-10 days). Treatment failure (defined as relapse of infection with the original microorganism) occurred in 2 cases (10.5%) during a median follow-up period of 3.9 years (range, 0.3-21.7 years). The 1-year cumulative risk of relapse was 11% (95% confidence interval, 0%-26%). Relapse of prosthetic joint infection due to penicillin-susceptible streptococci after debridement and retention of the prosthesis is uncommon. For patients who present with a well fixed prosthesis and a short duration of symptoms, debridement with retention appears to be an effective treatment modality. PMID- 12652385 TI - Ocular and respiratory symptoms attributable to inactivated split influenza vaccine: evidence from a controlled trial involving adults. AB - In 2000, an influenza vaccine was associated with unusual ocular and respiratory symptoms (known as "oculorespiratory syndrome" [ORS]) that possibly were due to numerous microaggregates of unsplit viruses present in the product. We assessed the potential for an improved vaccine formulation (for use in 2001-2002) to cause ORS and other symptoms in adults, using a double-blind, randomized, crossover study design. Symptoms were ascertained 24 h after 622 doses of vaccine and 626 doses of saline placebo were injected. The risk of ORS was 6.3% after vaccine injection and 3.4% after placebo injection, which yielded a significant vaccine attributable risk of 2.9% (95% confidence interval, 0.6-5.2). ORS symptoms were mild. Significant differences in risk after injection of vaccine versus placebo existed for ocular soreness and/or itching (2.4%), coughing (1.6%), and hoarseness (1.2%). Vaccine-attributable general symptoms were infrequent. We conclude that certain mild oculorespiratory symptoms were triggered by an influenza vaccine that was otherwise minimally reactogenic and, hence, that such symptoms might be associated with influenza vaccines in general. PMID- 12652386 TI - Risk factors for pediatric ventriculoperitoneal shunt infection and predictors of infectious pathogens. AB - Identification of risk factors for shunt infection and predictors of infectious pathogens may improve current methods to prevent and treat shunt infections. We reviewed data on 820 consecutive ventriculoperitoneal (VP) shunt placement procedures in 442 pediatric patients at our institution during 1992-1998. Ninety two shunts (11%) developed infection a median of 19 days (interquartile range, 11 35 days) after insertion. Premature birth (relative risk [RR], 4.81; 95% confidence interval [CI], 2.19-10.87), previous shunt infection (RR, 3.83; 95% CI, 2.40-6.13), and intraoperative use of the neuroendoscope (RR, 1.58; 95% CI, 1.01-2.50) were independent risk factors for shunt infection. The bacterial organisms early after shunt surgery (<14 days) were the same as those late after shunt surgery (>14 days). As determined by an analysis of the 92 infected shunts, hospital stay of >3 days at the time of shunt insertion (odds ratio [OR], 5.27; 95% CI, 1.15-25.3) and prior Staphylococcus aureus shunt infection (OR, 5.91; 95% CI, 1.35-25.9) independently increased the odds that S. aureus was the causal pathogen. PMID- 12652387 TI - Surgical site infections associated with methicillin-resistant Staphylococcus aureus: do postoperative factors play a role? AB - Patients with surgical site infections (SSIs) who underwent surgery during the period of September 1997 through December 1999 and January through July 2001 were retrospectively studied to compare patients infected with methicillin-resistant Staphylococcus aureus (MRSA) with those infected with organisms other than MRSA. Of patients with SSI who had known culture results, 77 (28.5%) of 270 had cultures that yielded MRSA. On univariate analysis, age of >or=70 years, duration of surgery of >or=4 h, duration of postoperative antibiotic treatment of >1 day, and discharge to a long-term care facility (LTCF) were significantly associated with MRSA SSI (P<.05 for all). On multivariate analysis, only discharge to an LTCF (odds ratio [OR], 2.3; P=.04) and duration of postoperative antibiotic treatment of >1 day (OR, 2.0; P=.03) were significantly associated with MRSA SSI; there was also a trend toward MRSA SSI being associated with use of a surgical drain for >1 day (P=.078). Postoperative factors may play a more important role in the causation of MRSA SSI than has previously been appreciated. PMID- 12652388 TI - Infectious disease outbreaks in nursing homes: an unappreciated hazard for frail elderly persons. AB - The common occurrence and dire consequences of infectious disease outbreaks in nursing homes often go unrecognized and unappreciated. Nevertheless, these facilities provide an ideal environment for acquisition and spread of infection: susceptible residents who share sources of air, food, water, and health care in a crowded institutional setting. Moreover, visitors, staff, and residents constantly come and go, bringing in pathogens from both the hospital and the community. Outbreaks of respiratory and gastrointestinal infection predominate in this setting, but outbreaks of skin and soft-tissue infection and infections caused by antimicrobial-resistant bacteria also occur with some frequency. PMID- 12652389 TI - Treatment and prevention of postherpetic neuralgia. AB - There have been 4 recent major advances in the treatment of postherpetic neuralgia (PHN) that are based on the results of randomized, controlled trials. These advances are the demonstrations that gabapentin, the lidocaine patch 5%, and opioid analgesics are efficacious in patients with PHN, and the report that nortriptyline and amitriptyline provide equivalent analgesic benefits for patients with PHN but that nortriptyline is better tolerated. The results of these clinical trials are briefly reviewed, and their implications for the treatment of patients with PHN are discussed. Despite these treatment advances, many patients remain refractory to current therapy, and the prevention of PHN has therefore become an important focus of current research. Research on administration of the varicella-zoster vaccine to prevent herpes zoster and on treatment of patients who have herpes zoster with combined antiviral and analgesic therapy to prevent PHN is discussed. PMID- 12652390 TI - Smallpox vaccination in 2003: key information for clinicians. PMID- 12652391 TI - Cryptosporidium species: new insights and old challenges. AB - Cryptosporidium species are protozoan parasites that cause mainly enteric illnesses in humans and other animals. The mode of transmission is most commonly waterborne, but other sources of infection, including foodborne and person-to person spread, have been documented. The environmental form of the parasite is resistant to most water purification methods, including chlorination. Cryptosporidium infection usually causes a self-limited diarrheal illness but can be life-threatening in immunocompromised individuals. There is no effective therapy for cryptosporidiosis. PMID- 12652392 TI - Prediction of coronary heart disease risk in HIV-infected patients with fat redistribution. AB - A metabolic syndrome has been described among human immunodeficiency virus (HIV) infected patients receiving highly active antiretroviral therapy; the syndrome is characterized by fat redistribution, insulin resistance, and dyslipidemia. We compared the 10-year coronary heart disease (CHD) risk estimates for 91 HIV infected men and women with fat redistribution with the risk estimates for 273 age-, sex-, and body mass index (BMI)-matched subjects enrolled in the Framingham Offspring Study. Thirty HIV-infected patients without fat redistribution were also compared with 90 age- and BMI-matched control subjects. The 10-year CHD risk estimate was significantly elevated among HIV-infected patients with fat redistribution, particularly among men; however, when they were matched with control subjects by waist-to-hip ratio, the 10-year CHD risk estimate did not significantly differ between groups. HIV-infected patients without fat redistribution did not have a greater CHD risk estimate than did control subjects. In addition, the CHD risk estimate was greatest in HIV-infected patients who had primary lipoatrophy, compared with those who had either lipohypertrophy or mixed fat redistribution. Therefore, although CHD risk is increased in HIV-infected patients with fat redistribution, the pattern of fat distribution and sex are potential important components in determining the risk in this population. PMID- 12652393 TI - Low incidence of community-acquired pneumonia among human immunodeficiency virus infected patients after interruption of Pneumocystis carinii pneumonia prophylaxis. AB - We compared the incidence of bacterial pneumonia among 336 patients who discontinued trimethoprim-sulfamethoxazole (TMP-SMX) as prophylaxis against Pneumocystis carinii pneumonia (PCP) with that among 75 patients who fulfilled the criteria for discontinuation but continued receiving prophylaxis. The difference in the overall incidence rates for the 2 groups (1.2 events per 100 person-years) was not statistically significant. Discontinuation of TMP-SMX prophylaxis against PCP is not associated with a significant increase in the incidence of bacterial pneumonia among patients with a sustained CD4 cell count increase to >200 cells/microL. PMID- 12652394 TI - Viral coinfections among African children infected with human immunodeficiency virus type 1. AB - City-dwelling children from Kenya who were infected with human immunodeficiency virus type 1 (HIV-1) were tested for coinfection with cytomegalovirus (CMV), human T cell lymphotropic viruses 1 and 2, Kaposi sarcoma-associated herpesvirus (KSHV), or hepatitis B, C, and G viruses. All children were found to be coinfected with CMV, whereas 5% had hepatitis G virus coinfection and 15% had KSHV coinfection. A protective role for hepatitis G virus cannot be excluded but likely affects only a minority of HIV-1-infected African children. PMID- 12652395 TI - Fungal peritonitis complicating peritoneal dialysis during an 11-year period: report of 46 cases. AB - The incidence of fungal peritonitis (FP) and the fungi that caused FP were evaluated in 422 patients treated with peritoneal dialysis. During an 11-year period, 804 episodes of peritonitis occurred, 46 (5.7%) of which were caused by fungi. Treatment was successful for 39 patients. Early diagnosis of FP and prompt therapy decreases morbidity and mortality. PMID- 12652396 TI - Trimethoprim-sulfamethoxazole as toxoplasmosis prophylaxis for heart transplant recipients. PMID- 12652398 TI - Putting salmonella contamination in perspective. PMID- 12652400 TI - A testament to sustained macrolide efficacy. PMID- 12652402 TI - P. carinii pneumonia in African children and the ineffectiveness of TMP-SMX prophylaxis. PMID- 12652404 TI - Autochthonous furuncular myiasis in the United States: case report and literature review. AB - Most cases of furuncular myiasis in the United States are acquired during travel to other countries. Autochthonous cases are infrequently reported and are most often due to accidental infestation of humans by larvae of flies belonging to the genus Cuterebra, commonly known as "rabbit bot flies" or "rodent bot flies." We describe a case of furuncular myiasis due to Cuterebra larvae in the hand of a patient residing in Wisconsin. The infestation was acquired while the patient gardened near a seepage lake during August. The patient developed papular lesions on the hand and received therapy for presumed bacterial furunculosis, which did not lead to improvement. Retrieval of one of the larvae from the patient permitted identification of the causative agent, and application of an occlusive ointment resulted in complete cure. North American cuterebrid myiasis is rare, and the diagnosis is frequently delayed. Myiasis should be considered when treating patients with refractory furunculosis. PMID- 12652405 TI - Brucella prosthetic joint infection: a report of 3 cases and a review of the literature. AB - We report 3 cases of Brucella melitensis infection of prosthetic hips and knees, and we summarize data about 4 cases reported in the literature. Six of the 7 affected patients were men. The median duration from prosthesis implantation to the onset of symptoms was 38.7 months. Five patients had only local symptoms. Preoperative joint aspirates yielded negative culture results for 3 patients, and blood culture results were negative for 6 patients. Excisional arthroplasty was the initial intervention for 3 patients. Three others responded well to medical therapy alone. One patient had relapse while receiving tetracycline and underwent total hip replacement. All patients were treated with combined antibiotic therapy for 6 weeks to 19 months. All had favorable long-term responses. The 3 patients we treated underwent a 2-staged resection arthroplasty. Antibiotics alone can be used to treat Brucella prosthetic joint infection, but loosening of the joint and clinical or microbiological failure must be treated with a 2-staged excisional arthroplasty and 3 months of treatment with doxycycline and rifampicin. PMID- 12652406 TI - Encephalitis associated with influenza B virus infection in 2 children and a review of the literature. AB - Two children with influenza B-associated encephalitis (IBAE) presented to our hospital during the winter of 2000-2001, both of whom had cases notable for mutism in association with encephalitis. A review of the literature identified 13 additional reports consistent with IBAE that contained sufficient data for analysis. Eleven of 15 reported cases occurred in children aged 95% donor engraftment by day 14 after transplantation. The incidence of Grade II to III acute graft-versus-host disease was 71%. There were two therapy related deaths. There were 8 partial responses and 3 complete responses (CRs) at day 28. Five additional CRs were observed at day 100 without withdrawal of cyclosporine or donor lymphocyte infusion. The rate of CRs for all 15 patients was 60%. The 1-year progression-free survival rate from time of study entry is 67% with only 1 relapse among 9 CRs. At a median potential follow-up of 28 months, the overall survival rate is 53%. These data demonstrate that a potent and durable graft-versus-lymphoma effect can occur against chemotherapy refractory lymphomas and suggest that this effect may be associated with rapid, complete donor chimerism after reduced-intensity alloHSCT. PMID- 12652468 TI - BEAM allogeneic transplantation for patients with Hodgkin's disease who relapse after autologous transplantation is safe and effective. AB - Because few patients failing autologous transplantation for Hodgkin's disease survive long-term, we explored reduced-intensity allografts using BEAM conditioning and early withdrawal of immunosuppression as an alternative to palliative chemotherapy. Ten patients with Hodgkin's disease underwent an allograft, receiving either matched sibling peripheral blood stem cells (5), partially matched sibling bone marrow (1), or matched unrelated bone marrow (4). Graft-versus-host disease (GVHD) prophylaxis was mini-methotrexate and FK-506 with weaning at day 60. The median age of patients was 35 years (range: 21 to 49 years). The median time from initial diagnosis was 73 months (range: 12 to 172 months) and from autograft was 49 months (range: 5 to 143 months). One patient was in CR, 5 patients were in partial remission, 3 were in relapse, and 1 patient had primary refractory disease. All patients' transplants engrafted rapidly, and the 100-day mortality was 0. Two patients developed acute GVHD. Five of the 9 patients beyond 100 days have developed mild chronic GVHD, of which 1 case was progressive and required systemic therapy. All 10 responded: 8 complete responses and 2 partial remissions. Three patients have relapsed (at 2, 6, and 8 months, respectively), 1 has died at 4 months. At a mean of 12 months (range: 1 to 21 months) after allograft, 9 of 10 patients are alive, with 7 in continuous remission. BEAM allogeneic transplantation with early reduction in immunosuppression is safe (no treatment-related deaths) and effective in advanced Hodgkin's disease where autografts have failed. A graft versus lymphoma effect appears to be a significant contributing factor in responding patients. PMID- 12652469 TI - 4-hydroperoxycyclophosphamide--purged peripheral blood stem cells for autologous transplantation in patients with acute myeloid leukemia. AB - We have performed a phase I dose escalation of 4-Hydroperoxycyclophosphamide (4HC) purging of autologous peripheral blood progenitor cells (PBPCs) to improve the outcome of autologous transplantation for patients with myeloid leukemia. Peripheral blood stem cells were mobilized after cytosine arabinoside of 2 g/m(2) every 12 hours x 8 doses with etoposide of 40 mg/kg total dose infused over 4 days, followed by growth factor support. The preparative regimen included Busulfan of 1 mg/kg orally every 6 hours x 16 doses, followed by etoposide of 60 mg/kg x 1 day (the patient with chronic myeloid leukemia received cyclophosphamide of 60 mg/kg/d x 2 days in lieu of etoposide). PBPCs purged with 4HC were infused following this induction. Toxicities included grade 3 or 4 skin rashes, stomatitis/mucositis, and delay in time to hematopoietic recovery. The maximum tolerated dose of 4HC used to purge PBPCs in this trial was 20 microg/mL, which resulted in an average of 18 days for white blood cells and 28 days for platelet recovery. With a median follow-up of 2.25 years in surviving patients, the 3-year disease free survival rate is 44% and the overall survival rate is 89%. These data suggest that autologous PBPCs are more sensitive than marrow purged with 4HC, tolerating less intense purging, although a survival advantage may still be seen and should be assessed in larger studies. Approaches to minimize stomatitis and protect normal stem cells from the toxicity of 4HC may improve the tolerance and efficacy of this approach. PMID- 12652470 TI - Lowered-intensity preparative regimen for allogeneic stem cell transplantation delays acute graft-versus-host disease but does not improve outcome for advanced hematologic malignancy. AB - Reduced conditioning intensity has extended the option of allogeneic hematopoietic stem cell transplantation to patients who cannot tolerate fully myeloablative regimens. However, relapse and graft-versus-host disease (GVHD) continue to be major causes of morbidity and mortality. We prospectively tested whether a moderate reduction of the intensity of the preparative regimen would lead to significant reduction in regimen-related toxicity without compromising tumor control in a cohort of 44 patients ineligible for conventional hematopoietic stem cell transplantation. Patients were conditioned with fludarabine, busulfan, mycophenolate, and total lymphoid irradiation. Tacrolimus and methotrexate were given as prophylaxis for GVHD. Donors were 5 of 6 or 6 of 6 matched family members. The median age was 61 years. Eleven patients had comorbid conditions that precluded conventional myeloablative transplantation. Fatal regimen-related organ toxicity occurred in 3 patients. The cumulative incidence of grade 2 to 4 or grade 3 to 4 acute GVHD by day 100 was 38% (95% confidence interval [CI] = 25%, 55%) and 20% (95% CI = 10%, 39%), respectively, with a median time to onset of 66 days. For the entire cohort, 1-year overall survival, disease-free survival, and relapse rates were 54% (95% CI = 41%, 71%), 47% (95% CI = 35%, 65%), and 37% (95% CI = 19%, 51%), respectively. Outcomes differed based on stage of disease at time of transplantation, advanced (n = 19) versus nonadvanced (n = 25). Median survival times were 138 days and 685 days for subjects with advanced and nonadvanced disease, respectively (P =.005). After adjusting for age and comorbidity, disease stage continued to be significantly associated with overall survival (P =.005). In conclusion, a moderate reduction in conditioning dose intensity resulted in delayed onset of acute GVHD (compared with historical controls). A reduction in conditioning intensity is associated with poor survival for patients with advanced-stage disease, highlighting the importance of the conditioning regimen for tumor control. PMID- 12652471 TI - Reconstitution of T-cell repertoire after autologous stem cell transplantation: influence of CD34 selection and cytomegalovirus infection. AB - The period of immunodeficiency following autologous hematopoietic stem cell transplantation is characterized by transient expansions of CD8+CD45RO+CD57+ T lymphocytes, displaying markers of an activated phenotype. Most evidence suggests that this early reconstitution results from proliferation of mature T cells that have survived conditioning or were transferred with the graft. Although homeostatic mechanisms are thought to act in maintaining total T-cell numbers, the degree to which antigen-driven expansions contribute and the nature of the stimulating antigens remain unclear. CD34 selection of stem cell grafts reduces the available T-cell pool, potentially delaying immune reconstitution and resulting in increased infective complications. In the allogeneic transplantation setting, lymphopenia has been associated with cytomegalovirus (CMV) infection risk and, if persistent, with adverse outcome. We prospectively studied patients undergoing CD34-selected (n = 13) or unselected (n = 13) autologous hematopoeitic stem cell transplantation for immune reconstitution and CMV infection. No significant differences were demonstrated between graft types with respect to lymphocyte subset recovery, T-cell receptor beta-chain variable region spectratype diversity, or CMV DNA detection rates (45% versus 40%). CMV infection was associated with a trend toward higher rather than lower CD8+ counts at 6 weeks posttransplantation (P =.08) that became significant by 3 months (P=.007), and that was associated with decreased T-cell receptor beta-chain variable region spectratype diversity (P =.01). CMV-specific HLA-tetramer analysis demonstrated transient expansions with CDR3 lengths corresponding to those of some of the major posttransplantation T-cell expansions demonstrated by spectratype analysis suggesting that CMV-specific T cells contribute to the pattern of immune reconstitution. PMID- 12652473 TI - Calorimetric investigation of anaerobic digestion: Biomass adaptation and temperature effect. AB - A high-resolution bench-scale calorimeter (Bio-RC1), specially suited for microbiological studies, has been used to investigate the response of an anaerobic granular sludge to repetitive glucose additions at various temperatures. Through the analysis of heat-evolution profiles resulting from substrate degradation, significant biological events have been identified: the feeble net heat uptake related to aceticlastic methanogenesis has been evaluated, and the detrimental effect of a nutrient deficiency, which led up to 75% decrease in heat production rate, has been detected. After several glucose pulses under non-limiting nutrient conditions, a reliable and reproducible reference thermal response, which is an important condition for the application of calorimetric measurements to detect inhibitory effects, was finally obtained. Afterward, glucose additions were repeated at various temperatures, and 57% biomass activity reduction, with respect to the reference value at 35 degrees C, was observed when temperature was fixed at 20 degrees C. Additionally, a preliminary toxicity test was also performed by exposing the biomass to increasing formaldehyde concentrations. PMID- 12652472 TI - Predictors of relapse and overall survival in Philadelphia chromosome-positive acute lymphoblastic leukemia after transplantation. AB - Allogeneic transplantation offers a potential cure for patients with Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ ALL). We performed a retrospective analysis examining pretransplantation and posttransplantation prognostic factors in 90 patients with Ph+ ALL. The median age of the patients was 33 years, with slightly more than half of the patients (58%) in clinical remission at the time of transplantation. Overall, patients had a nonrelapse mortality rate of 30%, a relapse percentage of 34%, and an estimated 5-year disease-free survival rate of 30%. Pretransplantation risk factors for relapse included the expression of the p190 transcript (relative risk [RR] = 5.1; P =.037), evidence of morphologic disease at the time of transplantation (RR = 3.9; P <.001), and type of donor (RR = 2.5; P =.015), with patients receiving autologous or matched related transplants having the highest risk of relapse. The detection of minimal residual disease by reverse transcription polymerase chain reaction for bcr-abl transcripts was a significant posttransplantation risk factor for relapse (RR = 4.4; P =.001), with posttransplantation patients expressing the p190 transcript having the highest risk of relapse (RR = 8.7; P =.0001). In addition, patients with chronic extensive graft-versus-host disease showed a significantly lower risk of relapse (RR = 0.33; P =.038). Thus, these findings indicate that several pretransplantation and posttransplantation risk factors exist for patients with Ph+ ALL. Together, these factors can be used to improve our risk stratification of patients with Ph+ ALL who undergo transplantation, which will greatly enhance our ability to counsel these patients and potentially lead to the development of more specific treatment plans for them. PMID- 12652474 TI - Impact of operating variables on the expanded bed adsorption of Saccharomyces cerevisiae cells using a concanavalin A derivatized perfluorocarbon. AB - The use of fluidizable affinity adsorbents for the adsorption of cells in expanded mode is investigated. Affinity adsorbents have been synthesized by immobilizing the lectin Concanavalin A onto the surface of triazine-activated perfluorocarbon-solids. The adsorbents were found to adsorb Saccharomyces cerevisiae cells from solution with adsorption capacities of up to 6.8 x 10(9) cells mL(-1). Adsorption kinetics were rapid with a time constant of 30 days after treatment) is a new phenomenon occurring after vascular brachytherapy. We report the analysis of 11 patients with late thrombosis after gamma-irradiation treatment of in-stent restenosis. All patients had in-stent restenosis and angina. Contributing factors to late thrombosis include long stents, small distal vessels, and complex lesion morphology. PMID- 12652495 TI - Coronary stenting in stable patients: identification of a low-risk subgroup that may not require adjunctive antiplatelet therapy. AB - The present study prospectively evaluated adjunctive antiplatelet therapy in patients without insulin-requiring diabetes during elective coronary stenting. Three hundred patients were randomized to one of three treatment groups: clopidogrel pretreatment, adjunctive abciximab, or control. Stenting was successful in 98% and no deaths occurred. Thirty-day and 1-year major adverse coronary events (MACEs) was similar in all groups. A subgroup of 109 patients undergoing single-vessel stenting of type A/B1 lesions with short guidewire times had no postprocedure myocardial infarction or 30-day MACE. We conclude that patients with these characteristics may safely undergo elective coronary stenting without adjunctive antiplatelet therapy. PMID- 12652496 TI - Mediastinal and neck hematoma after cardiac catheterization. AB - Vascular complications after cardiac catheterization are rare and usually occur at the access sites. However, vessels along the tract of the catheter can also be injured, causing bleeding and hematoma formation. We present a 57-year-old male who underwent cardiac catheterization via the radial approach, later developing neck and mediastinal hematoma, which was managed conservatively. This complication has only been reported once in the English literature. PMID- 12652498 TI - Eversion endarterectomy complicating radial artery access for left heart catheterization. AB - The use of the radial artery for arterial access for left heart catheterization is gaining popularity. We report the first case of traumatic eversion endarterectomy following the removal of the arterial sheath after the cannulation of the radial artery. The patient has no long-term vascular compromise of the limb and did not require surgical intervention. PMID- 12652497 TI - Intracoronary fracture and embolization of a coronary angioplasty balloon catheter: retrieval by a simple technique. AB - We report a technique for retrieval of a balloon along with a portion of its shaft from the coronary system using a simple technique that does not involve the use of snare or any other retrieval tool. An additional angioplasty wire and a balloon catheter were used to remove the balloon from the coronary system. PMID- 12652499 TI - Acute stent thrombosis after early withdrawal of platelet glycoprotein IIb/IIIa antagonists: potential rebound prothrombotic effect? AB - We report on two cases of acute coronary stent thrombosis after early withdrawal of different competitive inhibitors of the platelet glycoprotein IIb/IIIa receptor, eptifibatide and tirofiban. Differences in pharmacokinetics between different types of glycoprotein IIb/IIIa receptor blockers and a potential rebound prothrombotic effect with the use of these antiplatelet drugs are reviewed. PMID- 12652500 TI - Rotational and directional coronary atherectomy. PMID- 12652501 TI - Percutaneous occlusion of femoral artery pseudoaneurysm by para-aneurysmal saline injection. AB - The increasing number of coronary interventions and more aggressive anticoagulation is associated with a larger number of iatrogenic femoral pseudoaneurysms (PAs). The majority of PAs clot spontaneously or can be repaired by ultrasound-guided compression or ultrasound-guided thrombin injection. The therapeutic armamentarium for the remaining PAs ranges from different interventional procedures to surgical occlusion. In a small case series, we describe our initial experiences with a new, less invasive approach after unsuccessful compression therapy. Ultrasound-guided injection of 52 +/- 33 ml saline beneath the communication tract of the PA yields to rapid occlusion in six consecutive patients. During 4 weeks of follow-up, the PAs remained occluded in all patients and surgical intervention could be avoided. PMID- 12652502 TI - Thrombin injection for treatment of false aneurysms after failed compression therapy in patients on full-dose antiplatelet and heparin therapy. AB - The aim of this study was to gauge the effectiveness of thrombin injection after failed manual compression in patients with false aneurysms receiving full-dose antiplatelet and heparin therapy. In consecutive patients with failed manual compression therapy (ultrasound-guided manual compression, compression bandage, or both), thrombin was injected under ultrasound guidance. In 23 patients, thrombin was injected into the false aneurysm (100-2,000 units; mean, 895 +/- 520 units). All patients were on aspirin (median dose, 100 mg/day), clopidogrel (median dose, 75 mg/day), and either heparin (n = 3) with a partial thromboplastin time (PTT) > 55 sec or weight-adjusted enoxaparine (0.1 ml per 10 kg; maximum, 1 ml/b.i.d.). Twelve patients had received an i.v. GB IIIb/IIa antagonist 11-72 hr prior to diagnosis of the aneurysm. The mean width of the false aneurysms was 20.8 +/- 3.2 mm (range, 8.0-52.0 mm), length 29.7 +/- 32.6 mm (range, 9.0-147 mm), and depth 19 +/- 9.1 mm (range, 5.1-35.5 mm). Thrombosis after thrombin injection occurred in 21 patients within seconds. One patient required a second injection the next day, one patient underwent surgery. Overall success rate was 96%. No in-hospital complications occurred. In patients with false aneurysms and failed compression therapy under full-dose aspirin, clopidogrel, and heparin, selective thrombin injection is highly effective and safe. PMID- 12652503 TI - Stent angioplasty of severe atherosclerotic ostial renal artery stenosis in patients with diabetes mellitus and nephrosclerosis. AB - Atherosclerotic renal artery stenosis (ARAS) may lead to deterioration of renal function or hypertension. The clinical outcome after stent angioplasty of ARAS on renal function and blood pressure control in patients with diabetes and nephrosclerosis is the subject of some controversy. We have analyzed the results of our single-center experience with stent angioplasty for severe (>/= 70%) ostial ARAS and present here the results of a subgroup analysis of those patients who had diabetes mellitus and nephrosclerosis. From 1996 to 2001, 241 patients underwent stent angioplasty for the treatment of ARAS at our center. Of these, 99 patients had diabetes mellitus (41%) and 176 patients (73%) had nephrosclerosis defined as intrarenal resistance index (RI) >/= 0.7 diagnosed by duplex ultrasound. All lesions (n = 355) were treated successfully. Mean blood pressure at baseline was comparable and significantly improved immediately after the intervention in all groups (nondiabetics: 102 +/- 12 to 93 +/- 10 mm Hg; diabetics: 102 +/- 14 to 93 +/- 11 mm Hg; RI < 0.7: 105 +/- 13 to 95 +/- 10 mm Hg; RI = 0.7-0.8: 100 +/- 12 to 92 +/- 10 mm Hg; RI > 0.8: 102 +/- 15 to 92 +/- 11 mm Hg; P < 0.0001 each). Baseline serum creatinine was not significantly lower in nondiabetics compared to diabetics (1.46 +/- 0.9 vs. 1.62 +/- 1.2 mg %; P < 0.05) and increased in patients with nephrosclerosis (RI < 0.7: 1.18 +/- 0.6 mg %; RI = 0.7-0.8: 1.57 +/- 1.1 mg %; RI > 0.8: 1.96 +/- 1.6 mg %). Except for patients without nephrosclerosis who had a normal baseline creatinine, serum creatinine decreased significantly in all subgroups during follow-up. Stent angioplasty of ARAS offers favorable acute and long-term clinical results for the preservation of the renal function and for blood pressure control in patients with diabetes mellitus and nephrosclerosis. PMID- 12652504 TI - Carotid sinus reactions during carotid artery stenting: predictors, incidence, and influence on clinical outcome. AB - Carotid sinus reactions (CSR), defined as asystole >/= 3 sec and hypotension (systolic blood pressure ostial > isolated internal carotid artery; P < 0.001). The other independent predictors were presence of contralateral stenosis (P < 0.02), length of stenosis (P < 0.03), and balloon-to-artery ratio (P < 0.02). Occurrence of CSR was unrelated to periprocedural cerebral or cardiovascular complications (7.1% vs. 9.5%; NS). We conclude that CSR occurs frequently (40%) during CAS. Bifurcation location of stenosis is the most important predictor of CSR. CSR does not increase the risk of periprocedural complications. PMID- 12652505 TI - Cutting balloon angioplasty for undilatable venous stenoses causing dialysis graft failure. AB - Dialysis fistulas and bridge grafts frequently develop outflow stenoses in the venous limb, leading to graft failure. The venous stenoses are fibrotic and require percutaneous transluminal angioplasty with high-pressure balloons. We have encountered venous stenoses in 3 out of 219 hemodialysis patients with graft failure that were undilatable with 6-8 mm high-pressure balloons inflated to more than 24 atm. In each of these cases, treatment with a 4 mm cutting balloon successfully facilitated subsequent balloon dilation at lower pressures. PMID- 12652506 TI - Cavernous hemangioma of the foot and antecubital fossa: an alternative therapeutic option. AB - We present two cases of recurrent cavernous hemangiomas of the extremities. These conditions were initially treated unsuccessfully with surgical resection. After recurrence, the hemangiomas were treated by percutaneous transcatheter embolization. To the best of our knowledge, this is the first case report of treatment of cavernous hemangioma involving the extremities using an angiographically directed vascular embolization technique. PMID- 12652507 TI - Prevention of radiocontrast-induced nephropathy. PMID- 12652508 TI - Transthoracic left ventricular puncture for the assessment of patients with aortic and mitral valve prostheses: the Massachusetts General Hospital experience, 1989-2000. AB - Accurate assessment of suspected prosthetic valve dysfunction is critically important as reoperation carries high risk. Noninvasive methods of hemodynamic assessment of patients with both aortic and mitral mechanical valves continue to be frustrated by the interference created by prosthetic material and direct left ventricular puncture may be required for definitive hemodynamic assessment. We report the hemodynamic and angiographic results and outcomes of 38 consecutive patients with double valve replacement who underwent left ventricular puncture as part of evaluation of possible prosthetic dysfunction. These results were compared with those obtained by noninvasive testing. We found noninvasive assessment alone to be unsatisfactory as measurements of regurgitation and stenosis correlated poorly with those obtained by direct left ventricular puncture. Important information that altered patient management was obtained from invasive assessment in 68% of cases with an acceptable rate of complications. Therefore, hemodynamic and angiographic assessment using transthoracic left ventricular puncture should be entertained in patients with mitral and aortic valve replacement presenting with congestive heart failure and suspected prosthesis dysfunction. PMID- 12652509 TI - Simultaneous device closure of muscular ventricular septal defect and pulmonary valve balloon dilatation. AB - Percutaneous closure of ventricular septal defect and pulmonary valvuloplasty appears to be an attractive modality when compared to surgical treatment for both lesions. We report successful transcatheter closure of muscular ventricular septal defect with pulmonary valve balloon dilation performed simultaneously in a 27-year-old woman. Technical considerations in such a setting are being discussed. PMID- 12652510 TI - Partial anomalous pulmonary venous return: case report and review of the literature. AB - Isolated partial anomalous pulmonary venous return (APVR) is an uncommon finding. A patient with isolated APVR had pulmonary hypertension without demonstrable left to-right shunting prior to anticoagulant treatment of pulmonary emboli. After anticoagulant therapy, with a fall in pulmonary pressures and resistance to near normal levels, left-to-right shunting was then detected by oximetry and angiography. The anomaly was visualized on electron beam angiography and confirmed by conventional angiography after anticoagulant therapy. Contrary to the expected obligatory drainage of highly saturated blood associated with APVR, lack of detectable shunting was thought to be due to the obstruction of regional blood due to thromboembolism. PMID- 12652511 TI - The black box: what goes on inside the cardiac catheterization laboratory? PMID- 12652513 TI - Does it matter who you are or what you gain? An experimental study of preferences for resource allocation. AB - Using an experimental conjoint-analysis like approach, preferences for resource allocation were studied. An interactive survey was developed which was published in the World Wide Web. A convenience sample of undergraduate students participated in the study. Subjects were confronted with nine pairwise scenarios describing hypothetical patient groups in need of life-saving treatments. The patient groups presented differed in terms of their health-related lifestyle, socioeconomic status, age, life expectancy, quality of life after treatment and whether they had received extensive medical care in the past. Participants were asked to allocate a finite budget to each patient group. All attributes used in this study significantly influenced respondents' preferences on how to allocate the budget between patient groups. The general importance of attributes used in the QALY approach is supported by this study with quality of life being a central criterion. The distributional patterns observed were, however, different from those expected when rigorously adhering to the QALY framework: In only a very small fraction of allocations subjects distributed the entire budget strictly on the patient group expecting the highest QALY gain. The vast majority of responders was willing to trade efficiency for a more equal distribution of resources. The approach described can be used to analyze the importance people place on different attributes in resource allocation decisions and to study preferences for the final distribution of resources. PMID- 12652514 TI - Health status and heterogeneity of cost-sharing responsiveness: how do sick people respond to cost-sharing? AB - This paper examines whether the responsiveness of health care utilization to cost sharing varies by health status and the implications of such heterogeneity. First, we show theoretically that if health care utilization of those in poor health is less responsive to cost sharing, this, combined with the skewness of health expenditures in health status, leads to overestimates of the effect of cost sharing. This bias is exacerbated when elasticities are generalized to populations with greater expenditure skewness. Second, we show empirically that cost-sharing responsiveness does differ by health status using data from the Medicare Current Beneficiary Survey. Medicare beneficiaries are stratified into health status groups based on activity of daily living (ADL) impairments and self reported health status. Separately, for each of the health status groups, we estimate the effect of Medigap insurance on Part B utilization using a two-part expenditure model. We find that the change in expenditures associated with Medigap is smaller for those in poorer health. For example, when stratified using ADLs, Medigap insurance increases expenditures for 'healthy' groups by 36.4%, while the increase for the 'sick' group is 12.7%. Results are qualitatively the same for different forms of supplemental insurance and different methods of health status stratification. We develop a test to demonstrate that adjusting our results for selection bias would result in estimates of greater heterogeneity. Our results imply that a lowerbound estimate of the bias from neglecting heterogeneity is about 2-7%. PMID- 12652515 TI - Design techniques for stated preference methods in health economics. AB - This paper discusses different design techniques for stated preference surveys in health economic applications. In particular, we focus on different design techniques, i.e. how to combine the attribute levels into alternatives and choice sets, for choice experiments. Design is a vital issue in choice experiments since the combination of alternatives in the choice sets will determine the degree of precision obtainable from the estimates and welfare measures. In this paper we compare orthogonal, cyclical and D-optimal designs, where the latter allows expectations about the true parameters to be included when creating the design. Moreover, we discuss how to obtain prior information on the parameters and how to conduct a sequential design procedure during the actual experiment in order to improve the precision in the estimates. The designs are evaluated according to their ability to predict the true marginal willingness to pay under different specifications of the utility function in Monte Carlo simulations. Our results suggest that the designs produce unbiased estimations, but orthogonal designs result in larger mean square error in comparison to D-optimal designs. This result is expected when using correct priors on the parameters in D-optimal designs. However, the simulations show that welfare measures are not very sensitive if the choice sets are generated from a D-optimal design with biased priors. PMID- 12652516 TI - Revisiting the axiom of completeness in health care. AB - Experiments concerned with modelling individual preferences are based on the assumption of completeness i.e. it is assumed that individuals have well-defined preferences for any choice they are presented with. However, this may not be the case for goods such as health care, where individuals are not used to making choices. If this assumption is violated, the large body of experimental economic literature eliciting patient preferences in health care may be challenged. This paper reports the results of a discrete choice experiment carried out to examine the assumption of complete preferences within health care. The tests carried out are based on the comparison of preferences for three different goods for which different levels of formed preferences are expected: a supermarket; dentist consultation and bowel cancer screening. The results do not provide sufficient evidence to support this hypothesis. However, further research is required before these results are generalised. PMID- 12652517 TI - The changing public/private mix in dentistry in the UK--a supply-side perspective. AB - This paper examines the factors beyond NHS dentists' remuneration which may explain the variations in the public/private mix in general dental practitioners' workload in the UK. Given that NHS dentistry is subject to a fixed price system, the study focused mainly on non-income supply-side factors. Using data from a postal survey of a national random sample of dentists practising in the NHS in England in 1997, the study found that the strategies adopted by dentists in the management of NHS lists and the evidence that dentists spent significantly more time in private consultations compared with NHS consultations support concerns over equity of access to dental care and the quality of NHS dentistry. Dentists' attitudes to NHS and private dentistry revealed considerable ambiguity towards the NHS. While, on the one hand, the lack of sufficient demand for private dentistry emerged as a strong reason for remaining in the NHS, on the other, there was evidence that equity in access to dental treatment is still seen as an important principle. The implications of these findings in the context of recent discussion of the future of NHS dentistry are considered. PMID- 12652518 TI - Measuring the value of time for methadone maintenance clients: willingness to pay, willingness to accept, and the wage rate. AB - Three measures of the value of time - willingness to pay (WTP) for a reduction in travel time, willingness to accept (WTA) a monetary compensation to forgo it, and the wage rate - are evaluated and compared. WTP and WTA were estimated from the two-part regressions of time price, using contingent valuation methods with primary survey data. Systematic differences are found in comparisons of WTP and WTA with the wage rate. PMID- 12652520 TI - Regressive logistic and proportional hazards disease models for within-family analyses of measured genotypes, with application to a CYP17 polymorphism and breast cancer. AB - Various statistical methods have been proposed to evaluate associations between measured genetic variants and disease, including some using family designs. For breast cancer and rare variants, we applied a modified segregation analysis method that uses the population cancer incidence and population-based case families in which a mutation is known to be segregating. Here we extend the method to a common polymorphism, and use a regressive logistic approach to model familial aggregation by conditioning each individual on their mother's breast cancer history. We considered three models: 1) class A regressive logistic model; 2) age-of-onset regressive logistic model; and 3) proportional hazards familial model. Maximum likelihood estimates were calculated using the software MENDEL. We applied these methods to data from the Australian Breast Cancer Family Study on the CYP17 5'UTR T-->C MspA1 polymorphism measured for 1,447 case probands, 787 controls, and 213 relatives of case probands found to have the CC genotype. Breast cancer data for first- and second-degree relatives of case probands were used. The three methods gave consistent estimates. The best-fitting model involved a recessive inheritance, with homozygotes being at an increased risk of 47% (95% CI, 28-68%). The cumulative risk of the disease up to age 70 years was estimated to be 10% or 22% for a CYP17 homozygote whose mother was unaffected or affected, respectively. This analytical approach is well-suited to the data that arise from population-based case-control-family studies, in which cases, controls and relatives are studied, and genotype is measured for some but not all subjects. PMID- 12652521 TI - Optimal designs for estimating penetrance of rare mutations of a disease susceptibility gene. AB - Many clinical decisions require accurate estimates of disease risks associated with mutations of known disease-susceptibility genes. Such risk estimation is difficult when the mutations are rare. We used computer simulations to compare the performance of estimates obtained from two types of designs based on family data. In the first (clinic-based designs), families are ascertained because they meet certain criteria concerning multiple disease occurrences among family members. In the second (population-based designs), families are sampled through a population-based registry of affected individuals called probands, with oversampling of probands whose families are more likely to segregate mutations. We generated family structures, genotypes, and phenotypes using models that reflect the frequencies and penetrances of mutations of the BRCA1/2 genes. We studied the effects of risk heterogeneity due to unmeasured, shared risk factors by including risk variation due to unmeasured genotypes of another gene. The simulations were chosen to mimic the ascertainment and selection processes commonly used in the two types of designs. We found that penetrance estimates from both designs are nearly unbiased in the absence of unmeasured shared risk factors, but are biased upward in the presence of such factors. The bias increases with increasing variation in risks across genotypes of the second gene. However, it is small compared to the standard error of the estimates. Standard errors from population-based designs are roughly twice those from clinic-based designs with the same number of families. Using the root-mean-square error as a measure of performance, we found that in all instances, the clinic-based designs gave more accurate estimates than did the population-based designs with the same numbers of families. Rough variance calculations suggest that clinic-based designs give more accurate estimates because they include more identified mutation carriers. PMID- 12652522 TI - A score for Bayesian genome screening. AB - Bayesian Monte Carlo Markov chain (MCMC) techniques have shown promise in dissecting complex genetic traits. The methods introduced by Heath ([1997], Am. J. Hum. Genet. 61:748-760), and implemented in the program Loki, have been able to localize genes for complex traits in both real and simulated data sets. Loki estimates the posterior probability of quantitative trait loci (QTL) at locations on a chromosome in an iterative MCMC process. Unfortunately, interpretation of the results and assessment of their significance have been difficult. Here, we introduce a score, the log of the posterior placement probability ratio (LOP), for assessing oligogenic QTL detection and localization. The LOP is the log of the posterior probability of linkage to the real chromosome divided by the posterior probability of linkage to an unlinked pseudochromosome, with marker informativeness similar to the marker data on the real chromosome. Since the LOP cannot be calculated exactly, we estimate it in simultaneous MCMC on both real and pseudochromosomes. We investigate empirically the distributional properties of the LOP in the presence and absence of trait genes. The LOP is not subject to trait model misspecification in the way a lod score may be, and we show that the LOP can detect linkage for loci of small effect when the lod score cannot. We show how, in the absence of linkage, an empirical distribution of the LOP may be estimated by simulation and used to provide an assessment of linkage detection significance. PMID- 12652523 TI - Removing phenotypic distribution assumptions from tests of linkage disequilibrium for quantitative traits. AB - The transmission disequilibrium test (TDT) has become a family-based method of reference to search for linkage disequilibrium (LD). Although it was first developed for dichotomous traits, numerous approaches have extended the TDT to quantitative phenotypes that either rely on regression or variance component techniques. Both of these approaches are based on some phenotypic distribution assumptions, the violation of which can lead to inflation of type I error rates, and derive information from phenotypic variability, so that their power is very low under some selection schemes (e.g., one-tailed selection). We propose a new family-based test of association for quantitative traits, denoted maximum likelihood-binomial (MLB)-QTDT, which addresses the two previous issues by incorporating a latent binary variable that captures the LD information between the marker allele and the quantitative phenotype. The method can be understood as a classical TDT for binary traits that would include pure affected and pure unaffected children, and the probability for a child to be affected or unaffected depends on his/her quantitative phenotypic value. Simulation studies under the null hypothesis show that the MLB-QTDT provides very consistent type I errors even in small and/or selected samples. Under the alternative hypothesis, the MLB QTDT has good power to analyze one-tailed selected samples, and performs as well as a classical approach in other designs. The MLB-QTDT is a flexible distribution free method to test for LD with quantitative phenotypes in nuclear families, and can easily incorporate previous extensions developed in the context of family based association studies with binary traits. PMID- 12652524 TI - Does accounting for gene-environment (GxE) interaction increase the power to detect the effect of a gene in a multifactorial disease? AB - Despite tremendous efforts, few genes involved in the susceptibility for complex disorders have been identified. One explanation is that these disorders are a result of an interaction between genes and environment, and under such conditions, it may be difficult to measure the true genetic effect without accounting for the interaction. Umbach and Weinberg ([2000] Am. J. Hum. Genet. 66:251-261) proposed an association test which looks at the joint effects of genotype and environment, using case-parent trios. In this study, we explore under which conditions accounting for GxE interaction enhances one's ability to detect the role of genetic factors in complex diseases. Using asymptotic power calculations, we investigate the power to detect the gene effect over varying exposure frequencies and different scenarios of GxE interaction. We show that for a given sample size, interaction scenario, and allele frequency, the actual gain in power while accounting for the interaction depends on the magnitude of the exposure frequency: the largest gains are seen for relatively low exposure frequencies. Moreover, a loss of power can be observed when the exposure is frequent and/or the exposure effect is strong. If we consider a gene with a disease allele frequency of 0.2, with no effect in the absence of exposure, an exposure with a 10-fold increase risk and a GxE relative risk of 2, then when the exposure frequency is 0.1, accounting for GxE interaction increases the power to detect the gene effect in 200 trios by 10%; alternatively, when the exposure frequency is 0.9, it decreases the power by 15%. PMID- 12652525 TI - A general class of association tests for family-based data using weight functions. AB - Based on the symmetry of transmitted/nontransmitted alleles from heterozygous parents under the null hypothesis of no association, the work proposed here establishes a general statistical framework for constructing association tests with data from nuclear families with multiple affected children. A class of association tests is proposed for both diallelic and multiallelic markers. The proposed test statistics reduce to the transmission disequilibrium test for trios, to T(su) by Martin et al. ([1997] Am. J. Hum. Genet. 61:439-448) for affected sib pairs, and to the pedigree disequilibrium test by Martin et al. ([2000] Am. J. Hum. Genet. 67:146-154); [2001] Am. J. Hum. Genet. 68:1065-1067) when using affected sibships only. The association test used in simulation and for real data (sitosterolemia) is the one which has the best overall power in detecting association. This association test is generally more powerful than the association tests proposed by Martin et al. ([2000] Am. J. Hum. Genet. 67:146 154); [2001] Am. J. Hum. Genet. 68:1065-1067) when using only affected sibships. For the sitosterolemia data set, the association test has its most significant result (P-value=0.0012) for the marker locus on the same bacterial artificial chromosome as the disease locus. PMID- 12652526 TI - Kin-cohort evaluation of relative risks of genetic variants. AB - The kin-cohort study, an embedded retrospective cohort of first-degree relatives of probands, has been used frequently in conjunction with case-control studies to evaluate penetrance of high-risk genotypes. However, in principle, the kin-cohort approach can be used in conjunction with a case-control study in which family history data are carefully recorded to examine and validate the observed association of any candidate genotype with disease. Its utility for this purpose is governed by the relative efficiency of the kin-cohort analysis to test hypotheses about the relevant genotype-disease odds ratios, which in turn depends strongly on various factors, notably the magnitude of the disease incidence rate and the prevalence of the aberrant genotype. In this study, we compare the efficiencies of the two analytic approaches, case-control vs. kin-cohort, and illustrate the results in the context of cancer epidemiology. The kin-cohort analysis can provide substantial, important supplementary information for relatively common cancers such as breast, lung, colorectal, and prostate, and is also relatively efficient in general for the evaluation of the stronger and more rare genetic risk factors. Genet Epidemiol 24:220-229, 2003. PMID- 12652527 TI - Variants of developmental genes (TGFA, TGFB3, and MSX1) and their associations with orofacial clefts: a case-parent triad analysis. AB - We selected 262 case-parent triads from a population-based study of orofacial clefts in Norway, and examined variants of developmental genes TGFA, TGFB3, and MSX1 in the etiology of orofacial clefts. One hundred seventy-four triads of cleft lip cases (CL+/-P) and 88 triads of cleft palate only cases (CPO) were analyzed. There was little evidence for an association of any of these genes with CL+/-P. The strongest association was a 1.7-fold risk with two copies of the TGFB3-CA variant (95% CI=0.9-3.0). Among CPO cases, there was a 3-fold risk with two copies of the TGFA TaqI A2 allele, and no increase with one copy. Assuming this to be a recessive effect, we estimated a 3.2-fold risk among babies homozygous for the variant (95% CI=1.1-9.2). Furthermore, there was strong evidence of gene-gene interaction. While there was only a weak association of the MSX1-CA variant with CPO, the risk was 9.7-fold (95% CI=2.9-32) among children homozygous for both the MSX1-CA A4 allele and the TGFA A2 allele. No association of CPO with the TGFA variant was seen among the other MSX1-CA genotypes. In conclusion, no strong associations were found between CL+/-P and variants at these three genes. There was a possible recessive effect of the TGFA TaqI variant on the risk of CPO, with a 3-fold risk among children homozygous for the variant. The effect of this TGFA genotype was even stronger among children homozygous for the MSX1-CA A4 allele, raising the possibility of interaction between these two genes. PMID- 12652528 TI - Further evidence for a type 2 diabetes susceptibility locus on chromosome 11q. PMID- 12652529 TI - In vivo oxygen detection using exogenous hemoglobin as a contrast agent in magnetic resonance microscopy. AB - In this work we show that exogenous molecular hemoglobin (Hb) is an effective indicator of relative local oxygen tension in magnetic resonance (MR) microscopy studies in vivo. This approach is more sensitive than other MRI oximetry methods; it can be used at higher resolutions and in specimens with no blood oxygen level dependent (BOLD) effects. Using injection studies in flies, we show that Hb can permeate through relatively dense neural tissue, and that it is not obviously disruptive to physiology. Hb-injected flies show large changes in signal intensity (40-50%) when external O(2) levels are manipulated artificially from 0% to 21%. Oxygen-dependent contrast changes produced by exogenous Hb are detected in T(2)-weighted imaging experiments, and can be roughly calibrated if necessary. These studies demonstrate the feasibility of a contrast agent technique that may be useful for functional MRI (fMRI) studies of metabolism at tens of microns resolution. PMID- 12652530 TI - In vivo measurement of brain metabolites using two-dimensional double-quantum MR spectroscopy--exploration of GABA levels in a ketogenic diet. AB - A localized proton 2D double-quantum (DQ) spin-echo spectroscopy technique was implemented on 1.5 T clinical MRI scanners for the detection of gamma aminobutyrate (GABA) in the brain. The 2D approach facilitates separation of peaks overlapping with GABA in 1D DQ-filtered (DQF) spectra. This technique was applied to four normal adult volunteers and four children with intractable epilepsy. The coefficient of variation of the level of GABA and overlapping macromolecules at F2 = 3.0 ppm and F1 = 4.8 ppm was 0.08 in normal subjects. Three patients received 2D MRS scans before and after initiation of the ketogenic diet (KD): one patient showed a trend of decreasing GABA throughout the study, and two patients showed low initial GABA levels that increased over time. In addition to major metabolites and GABA, low-level metabolites (valine, leucine, and glutathione) were also identified in the 2D spectra. PMID- 12652531 TI - Relaxation times of 31P-metabolites in human calf muscle at 3 T. AB - Localized (31)P-STEAM experiments were performed at 3 T to estimate relaxation times of phosphorus-containing metabolites in the human calf muscle in vivo. T(1) and T(2) times of PCr, P(i), and NTPs were measured in the resting calf muscle of healthy subjects by varying TR and TE. The localization performance of the (31)P STEAM sequence was evaluated on a test object, resulting in a relative selection efficiency of 78 +/- 1% and contamination from outside the voxel of 0 +/- 2% under fully relaxed conditions. T(1) relaxation times (+/-SD, n = 5) of P(i), PCr, gamma-NTP, alpha-NTP, and beta-NTP obtained at 3 T are 5.2 +/- 1.0 s, 6.4 +/ 0.2 s, 4.5 +/- 0.3 s, 2.6 +/- 0.9 s, and 3.5 +/- 1.1 s, respectively. T(2) relaxation times (+/-SD, n = 6) of these metabolites are 148 +/- 17 ms, 334 +/- 30 ms, 78 +/- 13 ms, 55 +/- 7 ms, and 55 +/- 10 ms, respectively. Spin-lattice relaxation times established at 3 T are consistent with literature data at lower field strengths, whereas spin-spin relaxation times are lower. Several methodological considerations are discussed which may help improve quantification of metabolite concentrations in the human (calf) muscle in vivo by using localized noninvasive (31)P-MRS at 3 T, which is currently being tested for routine clinical applications. PMID- 12652532 TI - Muscle glycogenolysis is not activated by changes in cytosolic P-metabolites: a 31P and 1H MRS demonstration. AB - Skeletal muscle contraction and glycogenolysis are closely coupled. The standard explanation for this coupling, as taught in modern biochemistry textbooks, is that the metabolic products of contraction (ADP, AMP, P(i)) feed back to activate glycogenolytic enzymes, thus providing for resynthesis of ATP. However, both in vivo (31)P MRS analyses and chemical analyses of muscle extracts have provided results that are contrary to this theory, at least in its simplest form. The MRS studies suffer from ambiguous assumptions. More importantly, in (31)P MRS studies the dependent and independent variables are often confounded because the glycogenolytic rate is calculated from the same data which are used to calculate the other metabolic variables. The analysis of biopsies has been necessarily quite limited, and suffers from a different set of experimental artifacts. Thus, the problem of contraction-glycogenolysis-coupling was reassessed using a quantitatively accurate (1)H MRS method. It is confirmed that glycogenolysis and contractions are closely coupled during repetitive exercise, while glycogenolysis and P-metabolite concentrations are not. A simple metabolic feedback system cannot explain contraction-glycogenolysis-coupling. PMID- 12652533 TI - Detection of elevated glutathione in meningiomas by quantitative in vivo 1H MRS. AB - Glutathione has major roles in removing free radicals and toxins from normal tissues, but its presence in tumor cells hinders the effectiveness of many anticancer therapies. Analysis of short echo time brain tumor (1)H spectra at 1.5 T using a linear combination of metabolite spectra (LCModel) suggested a significant contribution of glutathione to meningioma spectra. By in vivo MRS (TE = 30 ms, TR = 2020 ms), reduced glutathione was found to be significantly elevated in meningiomas (3.3 +/- 1.5 mM, Mann Whitney, P < 0.005) compared to normal white matter (1.2 +/- 0.15 mM) and low-grade gliomas (1.0 +/- 0.26 mM), in agreement with published histofluorescence studies of tumor biopsies. Glx concentrations were also found to be elevated in meningiomas compared to astrocytomas or normal white matter, indicative of metabolic differences. The ability to noninvasively quantify reduced glutathione in vivo may aid selection of treatment therapies and also provide an indication of tumor aggressiveness. PMID- 12652534 TI - Is multicomponent T2 a good measure of myelin content in peripheral nerve? AB - Multicomponent T(2) relaxation of normal and injured rat sciatic nerve was measured. The T(2) relaxation was multiexponential, indicating the multicompartmental nature of T(2) decay in nerve tissue. The size of the short, observed T(2) component correlated very well with quantitative assessment of myelin using computer-assisted histopathological image analysis of myelin. Specifically, the size of the short T(2) component reflected the processes of myelin loss and remyelination accompanying Wallerian degeneration and regeneration following trauma. However, it represented all myelin present in the sample and did not distinguish between intact myelin and myelin debris. Other changes in T(2) spectra were also observed and could be correlated with axonal loss and inflammation. The study also questions the validity of previously offered interpretations of T(2) spectra of nerve. PMID- 12652535 TI - Cell internalization of anionic maghemite nanoparticles: quantitative effect on magnetic resonance imaging. AB - Anionic iron oxide nanoparticles are efficiently internalized into macrophages where they concentrate within micrometric endosomes, conferring on them a high magnetic susceptibility. The uptake of anionic maghemite nanoparticles by macrophages was quantified by an electron spin resonance (ESR) experiment. MR spin-echo sequences were performed with various TEs and TRs. The contrast enhancement was compared between two types of agarose phantoms with the same equivalent ferrite concentrations but containing either dispersed isolated nanoparticles or magnetically labeled macrophages. It is shown that the intracellular confinement of maghemite nanoparticles within micrometric endosomes results in a significant decrease of the longitudinal relaxivity and a moderate decrease of the transverse relaxivity compared to the relaxivities of the dispersed isolated nanoparticles. As a consequence, the signature of endosomal magnetic labeling consists of a negative contrast on T(1)-weighted images in the whole ferrite concentration range, whereas the presence of extracellular isolated nanoparticles can result in a positive enhancement. PMID- 12652536 TI - Spin-echo fMRI in humans using high spatial resolutions and high magnetic fields. AB - The Hahn spin-echo (HSE)-based BOLD effect at high magnetic fields is expected to provide functional images that originate exclusively from the microvasculature. The blood contribution that dominates HSE BOLD contrast at low magnetic fields (e.g., 1.5 T), and degrades specificity, is highly attenuated at high fields because the apparent T(2) of venous blood in an HSE experiment decreases quadratically with increasing magnetic field. In contrast, the HSE BOLD contrast is believed to arise from the microvasculature and increase supralinearly with the magnetic field strength. In this work we report the results of detailed and quantitative evaluations of HSE BOLD signal changes for functional imaging in the human visual cortex at 4 and 7 T. This study used high spatial resolution, afforded by the increased signal-to-noise ratio (SNR) of higher field strengths and surface coils, to avoid partial volume effects (PVEs), and demonstrated increased contrast-to-noise ratio (CNR) and spatial specificity at the higher field strengths. The HSE BOLD signal changes induced by visual stimulation were predominantly linearly dependent on the echo time (TE). They increased in magnitude almost quadratically in going from 4 to 7 T when the blood contribution was suppressed using Stejskal-Tanner gradients that suppress signals from the blood due to its inhomogeneous flow and higher diffusion constant relative to tissue. The HSE signal changes at 7 T were modeled accurately using a vascular volume of 1.5%, in agreement with the capillary volume of gray matter. Furthermore, high-resolution acquisitions indicate that CNR increased with voxel sizes < 1 mm(3) due to diminishing white matter or cerebrospinal fluid-space vs. gray matter PVEs. It was concluded that the high-field HSE functional MRI (fMRI) signals originated largely from the capillaries, and that the magnitude of the signal changes associated with brain function reached sufficiently high levels at 7 T to make it a useful approach for mapping on the millimeter to submillimeter spatial scale. PMID- 12652537 TI - Reproducibility study of 3D geometrical reconstruction of the human carotid bifurcation from magnetic resonance images. AB - The combined magnetic resonance imaging (MRI) and computational fluid dynamics (CFD) modeling approach is playing an increasingly important role in advancing our understanding of the relationship between hemodynamics and arterial disease. Nevertheless, such a modeling approach involves a number of uncertainties associated with various stages of the process. The present study is concerned with the reproducibility of geometry reconstruction, one of the most crucial steps in the modeling process. The reproducibility test was conducted on the right carotid bifurcation of eight normal human subjects, each of whom were scanned twice using the same MR protocol with an in-plane resolution of 0.625 mm. Models constructed from different scans of the same subject were compared and assessed using four quantitative measures: centerline distance, cross-sectional area, contour shape factors, and mean radius difference. The difference in the maximum carotid bulb area between the two scans was found to be <8.1% for all subjects. Shape factors (measuring the dissimilarity between two contours) of <10% were achieved in most of the common carotid arteries (CCAs) and internal carotid arteries (ICAs). The mean radius difference between the two scans was <0.4 mm for all subjects. Among the three vessels, the geometry of CCA was well reproduced by the reconstruction procedure in most of the cases, while the external carotid artery (ECA) showed the worst reproducibility. The impact of geometrical differences on CFD-predicted flow patterns and wall shear stress (WSS) will be investigated and discussed in a separate paper. PMID- 12652538 TI - Fast high-resolution 3D segmented echo planar imaging for dose mapping using a superheated emulsion chamber. AB - The superheated emulsion chamber (SEC) consists of superheated droplets of halocarbons in an aqueous gel. The gel resides in a pressure chamber. Brachytherapy sources can be implanted in the SEC for radiation dosimetry studies. Upon irradiation by ionizing radiation, the metastable droplets vaporize to form microbubbles. MRI can be used to determine the distribution of bubbles following irradiation of the SEC. In order to generate sufficient statistical accuracy in the determination of dose distributions around brachytherapy sources, it is necessary to use hundreds of irradiation cycles. Susceptibility-weighted images provide contrast between the gel and the vapor microbubbles. This article describes a 3D, blipped, double-sampled, segmented echo-planar imaging technique for rapidly imaging the SEC at 650 microm isotropic 3D resolution in about 2 min. This method was used with a pressure cycling SEC to acquire hundreds of images in several hours. Results are presented showing the 2D dose distribution generated by an (125)I source as measured in the SEC using this new imaging method. PMID- 12652539 TI - Quantitative tumor oxymetric images from 4D electron paramagnetic resonance imaging (EPRI): methodology and comparison with blood oxygen level-dependent (BOLD) MRI. AB - This work presents a methodology for obtaining quantitative oxygen concentration images in the tumor-bearing legs of living C3H mice. The method uses high resolution electron paramagnetic resonance imaging (EPRI). Enabling aspects of the methodology include the use of injectable, narrow, single-line triaryl methyl spin probes and an accurate model of overmodulated spectra. Both of these increase the signal-to-noise ratio (SNR), resulting in high resolution in space (1 mm)(3) and oxygen concentrations (approximately 3 torr). Thresholding at 15% the maximum spectral amplitude gives leg/tumor shapes that reproduce those in photographs. The EPRI appears to give reasonable oxygen partial pressures, showing hypoxia (approximately 0-6 torr, 0-10(3) Pa) in many of the tumor voxels. EPRI was able to detect statistically significant changes in oxygen concentrations in the tumor with administration of carbogen, although the changes were not increased uniformly. As a demonstration of the method, EPRI was compared with nearly concurrent (same anesthesia) T(2)*/blood oxygen level-dependent (BOLD) MRI. There was a good spatial correlation between EPRI and MRI. Homogeneous and heterogeneous T(2)*/BOLD MRI correlated well with the quantitative EPRI. This work demonstrates the potential for EPRI to display, at high spatial resolution, quantitative oxygen tension changes in the physiologic response to environmental changes. PMID- 12652540 TI - Capillarization of the sinusoids in liver fibrosis: noninvasive assessment with contrast-enhanced MRI in the rabbit. AB - Sinusoidal capillarization induces microcirculatory changes in liver cirrhosis and fibrosis. The purpose of this study was to assess whether contrast-enhanced MRI can be used to demonstrate the effects of sinusoidal capillarization in liver fibrosis. Dynamic MRI after injection of a low-molecular-weight contrast agent of 0.56 kDa (Gd-DOTA), and two high-molecular-weight contrast agents of 6.47 kDa and 52 kDa (P792 and P717) was performed in rabbits with liver fibrosis induced by cholesterol and diethylstilbestrol. The hepatic distribution volume accessible to the high-molecular-weight agents decreased in the rabbits with liver fibrosis (P792: 7.8% +/- 1.7% vs. 10.1% +/- 1.8% in normal rabbits, P =.038; P717: 6.2% +/ 2.1% vs. 9.7% +/- 1.6% in normal rabbits, P =.007), whereas the hepatic mean transit time (MTT) of the low-molecular-weight agent was increased (15.9 +/- 8.0 s vs. 8.8 +/- 2.6 s in normal rabbits, P =.015). In rabbits with liver fibrosis, the clearance of indocyanine green (ICG) was correlated with the volume accessible to the high-molecular-weight agents (P792: r = 0.810, P =.015; P717: r = 0.857, P =.007). The collagen content of the liver was inversely correlated with the distribution volume of P717 (r = -.833, P =.010) and with the ICG clearance (r = -.810, P =.015). It was concluded that the microcirculatory changes induced by sinusoidal capillarization in liver fibrosis can be demonstrated noninvasively with MRI. PMID- 12652541 TI - Comparison of new sequences for high-resolution cartilage imaging. AB - The high prevalence of osteoarthritis continues to demand improved accuracy in detecting cartilage injury and monitoring its response to different treatments. MRI is the most accurate noninvasive method of diagnosing cartilage lesions. However, MR imaging of cartilage is limited by scan time, signal-to-noise ratio (SNR), and image contrast. Recently, there has been renewed interest in SNR efficient imaging sequences for imaging cartilage, including various forms of steady-state free-precession as well as driven-equilibrium imaging. This work compares several of these sequences with existing methods, both theoretically and in normal volunteers. Results show that the new steady-state methods increase SNR efficiency by as much as 30% and improve cartilage-synovial fluid contrast by a factor of three. Additionally, these methods markedly decrease minimum scan times, while providing 3D coverage without the characteristic blurring seen in fast spin-echo images. PMID- 12652542 TI - Phased-array MRI of canine prostate using endorectal and endourethral coils. AB - A four-channel phased array consisting of one surface coil, two endorectal coils, and one flexible endourethral loop coil was designed for MRI of the canine prostate. The endorectal coils provide high signal in the posterior region of the prostate, while the endourethral and surface coils are sensitive to the central and anterior regions of the prostate. Gel phantom experiments indicate that the proposed phased-array configuration generates 15 times more signal-to-noise ratio (SNR) than a combination of two surface coils and one endorectal coil within the posterior region of the prostate; the performance of the two configurations is comparable near the anterior prostate surface. Ultimate intrinsic SNR (UISNR) analysis was used to compare the proposed phased array's performance to the best possible SNR for external coils. This analysis showed that the proposed phased array outperforms the best-case external coil within the posterior and central regions of the prostate by up to 20 times. In canine experiments in vivo, high resolution fast spin-echo (FSE) images of the prostate were obtained with a pixel size of 230 microm obtained in 3 min 12 s. The proposed phased-array design potentially can be used to increase the accuracy of prostate cancer staging and the feasibility of MR-guided prostate interventions. PMID- 12652543 TI - Classification and quantification of neuronal fiber pathways using diffusion tensor MRI. AB - Quantitative characterization of neuronal fiber pathways in vivo is of significant neurological and clinical interest. Using the capability of MR diffusion tensor imaging to determine the local orientations of neuronal fibers, novel algorithms were developed to bundle neuronal fiber pathways reconstructed in vivo with diffusion tensor images and to quantify various physical and geometric properties of fiber bundles. The reliability of the algorithms was examined with reproducibility tests. Illustrative results show that consistent physical and geometric measurements of novel properties of neuronal tissue can be obtained, which offer considerable potential for the quantitative study of fiber pathways in vivo. PMID- 12652544 TI - Steady-state free precession with myocardial tagging: CSPAMM in a single breathhold. AB - A method is presented that combines steady-state free precession (SSFP) cine imaging with myocardial tagging. Before the tagging preparation at each ECG-R wave, the steady-state magnetization is stored as longitudinal magnetization by an alpha/2 flip-back pulse. Imaging is continued immediately after tagging preparation, using linearly increasing startup angles (LISA) with a rampup over 10 pulses. Interleaved segmented k-space ordering is used to prevent artifacts from the increasing signal during the LISA rampup. First, this LISA-SSFP method was evaluated regarding ghost artifacts from the steady-state interruption by comparing LISA with an alpha/2 startup method. Next, LISA-SSFP was compared with spoiled gradient echo (SGRE) imaging, regarding tag contrast-to-noise ratio and tag persistence. The measurements were performed in phantoms and in six subjects applying breathhold cine imaging with tagging (temporal resolution 51 ms). The results show that ghost artifacts are negligible for the LISA method. Compared to the SGRE reference, LISA-SSFP was two times faster, with a slightly better tag contrast-to-noise. Additionally, the tags persisted 126 ms longer with LISA-SSFP than with SGRE imaging. The high efficiency of LISA-SSFP enables the acquisition of complementary tagged (CSPAMM) images in a single breathhold. PMID- 12652545 TI - Local reconstruction of stenosed sections of artery using multiple MRA acquisitions. AB - A method for reconstructing magnetic resonance angiography (MRA) volumes from successive acquisitions is described. The method is based on double oblique acquisitions of highly anisotropic MRA volumes, each of which corresponds to reduced k-space filling. These partial k-spaces are then combined to obtain a 3D k-space adapted to the frequency spread of the angiographic image of the stenosis. The SNR-resolution compromise of MRA is thus improved by focusing the acquisition on the most relevant k-space regions. The reconstruction is performed directly in k-space by averaging the partial k-spaces. The feasibility of the method was demonstrated in studies on a Lucite stenosis phantom, on MRAs of carotid arteries using three bolus injections, and on MRAs of renal arteries using a single contrast injection. PMID- 12652546 TI - Unsupervised reconstruction of a three-dimensional left ventricular strain from parallel tagged cardiac images. AB - A new algorithm, called the Unsupervised Tag ExTraction and Heart strain(E) Reconstruction (UNTETHER) algorithm, is presented for quantifying three dimensional (3D) myocardial strain in tagged cardiac MR images. Five human volunteers and five postinfarct patients were imaged. 3D strains measured by UNTETHER and a user-supervised technique were compared. Each study was analyzed in 49 +/- 8 min with UNTETHER, compared to approximately 4 hr with the user supervised technique. For pooled human data, the correlation coefficient between the two methods for circumferential shortening (E(cc)) was r = 0.91 at the mid wall (P < 0.0005). UNTETHER is capable of measuring wall motion abnormalities resulting from coronary artery disease, and has the potential to overcome the main limitations (time and user-supervision requirements) to routine clinical use of tagged cardiac MRI. PMID- 12652547 TI - Indirect imaging of ethanol via magnetization transfer at high and low magnetic fields. AB - Ethanol (EtOH) is believed to exert its neurochemical effects through interactions with brain cellular components, which causes a fraction of brain EtOH to have a lower molecular mobility. This facilitates magnetization transfer to other molecules similarly associated with macromolecules, such as water. It was hypothesized that this effect can be used in vivo to image EtOH indirectly via the much stronger brain tissue water resonance. EtOH-containing bovine serum albumin samples were used to demonstrate magnetic coupling between EtOH and water at 7 T and 1.5 T. Spectroscopy and imaging experiments demonstrated that EtOH signal saturation yielded greater water signal reduction than inversion and that this reduction scaled with EtOH concentration in the BSA samples. In human brain at physiologically relevant brain EtOH concentrations, water signal reductions were measurable when saturating the EtOH resonance. Strengths and limitations of indirectly imaging brain EtOH are discussed. PMID- 12652548 TI - Assessment of global and regional myocardial function in the mouse using cine and tagged MRI. AB - Mouse models are expected to play an important role in future investigations of human cardiac diseases. In the present report, MRI methods for determining global and regional cardiac function in the mouse are demonstrated. ECG-gated cine images were acquired in five C57BL/6 mice at physiological temperatures (37 degrees C) and heart rates of 500 +/- 50 beats per minute. Left ventricular mass, ejection fraction, and cardiac output were estimated from the resulting images. Regional myocardial function was also determined in three animals by application of 2D SPAtial Modulation of Magnetization (SPAMM) in combination with the cine protocol. The quality of the tagged images was sufficient to allow mapping of myocardial strains and displacements. The results of the regional strain analysis were consistent with similar studies in larger animals. This work demonstrates the first characterization of regional myocardial function in the mouse via SPAMM techniques. PMID- 12652549 TI - High-resolution in vivo CBV mapping with MRI in wild-type mice. AB - NMR microimaging has the potential to elucidate cerebrovascular abnormalities in mouse models. In this study, the relative regional cerebral blood volume (CBV) map is presented for C57BL6/J wild-type mice. The CBV mapping was based on changes in the steady-state NMR transverse relaxation rate (DeltaR(2)) associated with the presence of a superparamagnetic intravascular contrast agent (MION) with a long blood halflife. The experiments were performed at 9.4 T at a voxel size of 100 microm x 100 microm x 600 microm. Fine details, such as the hippocampal and olfactory bulb area, were visualized in the CBV map. The relative regional CBV values of various brain regions were measured. The DeltaR(2) dosage dependency and MION tissue clearance in mouse are also reported. PMID- 12652550 TI - An analytical solution for the SSFP signal in MRI. AB - Among previous analyses of the steady-state free-precession (SSFP) signal in rapid MRI, one treatment resulted in equations that require the evaluation of infinite binomial series. Here, an analytical solution is derived by a transformation into the power series expansion of the derivative of the inverse sine function, which is essentially a root. The treatment is extended to include higher-order signals. The results demonstrate the identity of the vastly different equations for the SSFP signals reported so far. Applications consist of the derivation of closed expressions for the signal in echo-shifted MRI and a corresponding analysis of TrueFISP sequences. PMID- 12652551 TI - Novel T*2-weighted contrast preparation scheme for segmented k-space myocardial imaging. AB - A novel T*(2)-weighted contrast-preparation scheme is described for use with segmented k-space cardiac sequences. This approach frees the imaging phase from the requirement of a long TE and, hence, a relatively long TR. A [90 degrees (x) tau-90 degrees (rho)] preparation scheme is used to acquire four image data sets with the phase rho of the second pulse set to x, y, -x, and -y. The rho = x raw data is subtracted from the rho = -x data to form the "x" image, with a similar subtraction to generate the "y" image. These images are added in quadrature to obtain the T*(2)-weighted image. The method results in reduced artifact compared to a simple two-image scheme with rho = x, and y. T*(2) was measured in the myocardial septum in six normal volunteers by comparing tau = 7 and 28 ms images, and it was found to be 44 +/- 5 ms at 0.95 T. PMID- 12652552 TI - On the transient phase of balanced SSFP sequences. AB - The signal intensity of balanced steady-state free precession (SSFP) imaging is a function of the proton density, T(1), T(2), flip angle (alpha), and repetition time (TR). The steady-state signal intensity that is established after about 5*T(1)/TR can be described analytically. The transient phase or the approach of the echo amplitudes to the steady state is an exponential decay from the initial amplitude after the first excitation pulse to the steady-state signal. An analytical expression of the decay rate of this transient phase is presented that is based on a simple analysis derived from the Bloch equations. The decay rate is a weighted average of the T(1) and T(2) relaxation times, where the weighting is determined by the flip angle of the excitation pulses. Thus, balanced SSFP imaging during the transient phase can provide various contrasts depending on the flip angle and the number of excitation pulses applied before the acquisition of the central k-space line. In addition, transient imaging of hyperpolarized nuclei, such as (3)He, (129)Xe, or (13)C, can be optimized according to their T(1) and T(2) relaxation times. PMID- 12652553 TI - Analysis of associations with change in a multivariate outcome variable when baseline is subject to measurement error. AB - A simple general algorithm is described for correcting for bias caused by measurement error in independent variables in multivariate linear regression. This algorithm, using standard software, is then applied to several approaches to the analysis of change from baseline as a function of baseline value of the outcome measure plus other covariates, any of which might have measurement error. The algorithm may also be used when the independent variables differ by component of the multivariate independent variable. Simulations indicate that under various conditions bias is much reduced, as is mean squared error, and coverage of 95 per cent confidence intervals is good. PMID- 12652554 TI - Power and sample size calculations for generalized regression models with covariate measurement error. AB - Covariate measurement error is often a feature of scientific data used for regression modelling. The consequences of such errors include a loss of power of tests of significance for the regression parameters corresponding to the true covariates. Power and sample size calculations that ignore covariate measurement error tend to overestimate power and underestimate the actual sample size required to achieve a desired power. In this paper we derive a novel measurement error corrected power function for generalized linear models using a generalized score test based on quasi-likelihood methods. Our power function is flexible in that it is adaptable to designs with a discrete or continuous scalar covariate (exposure) that can be measured with or without error, allows for additional confounding variables and applies to a broad class of generalized regression and measurement error models. A program is described that provides sample size or power for a continuous exposure with a normal measurement error model and a single normal confounder variable in logistic regression. We demonstrate the improved properties of our power calculations with simulations and numerical studies. An example is given from an ongoing study of cancer and exposure to arsenic as measured by toenail concentrations and tap water samples. PMID- 12652555 TI - The use of regression models for medians when observed outcomes may be modified by interventions. AB - Some outcomes used in epidemiological studies and clinical trials are prone to modification by interventions, for example, individuals with high blood pressure are likely to receive antihypertensive medication. Often the scientific interest is in the relationship of covariates (exposures or randomized treatment) to the outcomes that would have been observed in the absence of intervention. We compare three approaches to the analysis of such data: ignoring the intervention; excluding individuals who receive the intervention, and assuming that individuals who receive the intervention have underlying outcomes above the median. The latter approach requires comparison of median outcomes between groups. In many situations it is plausible that neither the probability of intervention nor the effect of intervention depend on the covariates. In this case we show that analysis of medians is unbiased in general, that the other approaches are biased towards the null but that ignoring the intervention typically has the greatest power for detecting an effect. In other situations, ignoring the intervention and excluding individuals who receive the intervention may be biased towards or away from the null, and we recommend analysis of medians. We illustrate practical analysis of medians in a study of the association between adult blood pressure and birth weight. Adjustment for confounders is performed by median regression. We show that the significance levels are comparable to those derived from logistic regression. We also discuss the effect of grouping of blood pressure and the need for bootstrap standard errors. PMID- 12652556 TI - An EM-based semi-parametric mixture model approach to the regression analysis of competing-risks data. AB - We consider a mixture model approach to the regression analysis of competing risks data. Attention is focused on inference concerning the effects of factors on both the probability of occurrence and the hazard rate conditional on each of the failure types. These two quantities are specified in the mixture model using the logistic model and the proportional hazards model, respectively. We propose a semi-parametric mixture method to estimate the logistic and regression coefficients jointly, whereby the component-baseline hazard functions are completely unspecified. Estimation is based on maximum likelihood on the basis of the full likelihood, implemented via an expectation-conditional maximization (ECM) algorithm. Simulation studies are performed to compare the performance of the proposed semi-parametric method with a fully parametric mixture approach. The results show that when the component-baseline hazard is monotonic increasing, the semi-parametric and fully parametric mixture approaches are comparable for mildly and moderately censored samples. When the component-baseline hazard is not monotonic increasing, the semi-parametric method consistently provides less biased estimates than a fully parametric approach and is comparable in efficiency in the estimation of the parameters for all levels of censoring. The methods are illustrated using a real data set of prostate cancer patients treated with different dosages of the drug diethylstilbestrol. PMID- 12652557 TI - Regression analysis with multiplicative and time-varying additive regression coefficients with examples from breast and colon cancer. AB - Regression analysis may be used to simplify the representation of mortality rates when there are many significant prognostic covariates or to adjust for confounding effects. The principal request of the regression model in this range of use is to have unbiased parameter estimates. A model with constant multiplicative and time-varying additive regression coefficients is discussed. The model allows some covariate effects to be multiplicative while allowing others to have a time-varying additive effect. Thus, it is a mix of classical Cox regression and Aalen's additive risk model. A major characteristic of cancer mortality rates, in contrast to general mortality rates, is that hazard rates, after a potentially initial increase, decrease, although not always tending to zero. Cancer diseases, like breast and colon cancer, have significantly increased cause-specific mortality rates even 20 years after diagnosis. Another major feature in cancer survival analysis is that many covariate effects are time varying. Some covariate effects, like age at diagnosis, may only be significant for a limited time after diagnosis. Furthermore, some treatment procedures may initially decrease the mortality, while the long-term effect may be opposite. A third issue is that average covariate effects are very often not multiplicative. Estimation is carried out iteratively; the cumulative additive regression functions are estimated non-parametrically using a least-squares method and the multiplicative parameters are estimated from the partial likelihood. The method is applied on 3201 female breast cancer and 1372 male colon cancer patients. PMID- 12652558 TI - Truncated negative binomial mixed regression modelling of ischaemic stroke hospitalizations. AB - A zero-truncated negative binomial mixed regression model is presented to analyse overdispersed positive count data. The study is motivated by the determination of pertinent risk factors associated with ischaemic stroke hospitalizations. Random effects are incorporated in the linear predictor to adjust for inter-hospital variations and the dependency of clustered observations using the generalized linear mixed model approach. The method assists hospital administrators and clinicians to estimate the number of subsequent readmissions based on characteristics of the patient at the index stroke. The findings have important implications on resource usage, rehabilitation planning and management of acute stroke care. PMID- 12652559 TI - Transition models for change-point estimation in logistic regression. AB - Although a wide variety of change-point models are available for continuous outcomes, few models are available for dichotomous outcomes. This paper introduces transition methods for logistic regression models in which the dose response relationship follows two different straight lines, which may intersect or may present a jump at an unknown change-point. In these models, the logit includes a differentiable transition function that provides parametric control of the sharpness of the transition at the change-point, allowing for abrupt changes or more gradual transitions between the two different linear trends, as well as for estimation of the location of the change-point. Linear-linear logistic models are particular cases of the proposed transition models. We present a modified iteratively reweighted least squares algorithm to estimate model parameters, and we provide inference procedures including a test for the existence of the change point. These transition models are explored in a simulation study, and they are used to evaluate the existence of a change-point in the association between plasma glucose after an oral glucose tolerance test and mortality using data from the Mortality Follow-up of the Second National Health and Nutrition Examination Survey. PMID- 12652561 TI - Regression analysis of incomplete medical cost data. AB - The accumulation of medical cost over time for each subject is an increasing stochastic process defined up to the instant of death. The stochastic structure of this process is complex. In most applications, the process can only be observed at a limited number of time points. Furthermore, the process is subject to right censoring so that it is unobservable after the censoring time. These special features of the medical cost data, especially the presence of death and censoring, pose major challenges in the construction of plausible statistical models and the development of the corresponding inference procedures. In this paper, we propose several classes of regression models which formulate the effects of possibly time-dependent covariates on the marginal mean of cost accumulation in the presence of death or on the conditional means of cost accumulation given specific survival patterns. We then develop estimating equations for these models by combining the approach of generalized estimating equations for longitudinal data with the inverse probability of censoring weighting technique. The resultant estimators are shown to be consistent and asymptotically normal with simple variance estimators. Simulation studies indicate that the proposed inference procedures behave well in practical situations. An application to data taken from a large cancer study reveals that the Medicare enrollees who are diagnosed with less aggressive ovarian cancer tend to accumulate medical cost at lower rates than those with more aggressive disease, but tend to have higher lifetime costs because they live longer. PMID- 12652560 TI - Dynamic Cox modelling based on fractional polynomials: time-variations in gastric cancer prognosis. AB - The most popular model used for survival analysis is the proportional hazards regression model proposed by Cox. This is mainly due to its exceptional simplicity. Nevertheless the fundamental assumption of the Cox model is the proportionality of the hazards. For many applications, however, this assumption is doubtful. Proposals to extend the Cox model for non-proportional hazards to allow for dynamic effect structures usually either depend on prespecifications or require non-standard estimation techniques and are thus not favoured in application. Moreover, tests to verify the dynamic effect structures are not straightforward or lack omnibus power. In this paper we propose a flexible method for modelling dynamic effects in survival data within the Cox framework. The method is based on fractional polynomials as introduced by Royston and Altman. This allows for a transformation of the dynamic predictor which leads back to the conventional Cox model and hence fitting is straightforward using standard estimation techniques. In addition, it offers the possibility of easily verifying the existence of time-variation. We describe a model selection algorithm which selects time-varying effects only when evidence is given in the data. We illustrate the properties of the approach in a simulation study and compare it with other methods. In a survival study of gastric cancer patients, we apply the approach to analyse dynamic structures in the effects of prognostic factors. PMID- 12652562 TI - The predictive value of microbiologic diagnostic tests if asymptomatic carriers are present. Ronny K. Gunnarsson and Jan Lanke, Statistics in Medicine 2002; 21:1773-1785. PMID- 12652563 TI - Spatial mixture relative risk models applied to disease mapping. A. B. Lawson and A. Clark, Statistics in Medicine 2002; 21:359-370. PMID- 12652565 TI - Standardization using the harmonically weighted ratios: internal and external comparisons. Wen-Chung Lee, Statistics in Medicine 2002; 21: 247-261. PMID- 12652567 TI - Protein fractionation in a multicompartment device using Off-Gel isoelectric focusing. AB - A new protein fractionation technique based on off-gel isoelectric focusing (IEF) is presented, where the proteins are separated according to their isoelectric point (pI) in a multiwell device with the advantage to be directly recovered in solution for further analysis. The protein fractions obtained with this technique have then been characterized with polymer nanoelectrospray for mass spectrometry (MS) analyses or with Bioanalyzer for mass identification. This methodology shows the possibility of developing alternatives to the classical two-dimensional (2-D) gel electrophoresis. One species numerical simulation of the electric field distribution during off-gel separation is also presented in order to demonstrate the principle of the purification. Experiments with pI protein markers have been carried out in order to highlight the kinetics and the efficiency of the technique. Moreover, the resolution of the fractionation was shown to be 0.1 pH unit for the separation of beta-lactoglobulin A and B. In addition, the isoelectric fractionation of an Escherichia coli extract was performed in standard solubilization buffer to demonstrate the performances of the technique, notably for proteomics applications. PMID- 12652568 TI - Poly(vinyl alcohol) hydrogels: their synthesis and steps towards control of electroendosmosis. AB - Poly(vinyl alcohol) (PVAl) hydrogel networks cross-linked with glutaraldehyde were prepared and their properties as membranes examined using a variety of techniques including preparative electrophoresis. Electroendosmosis (EEO) was observed and shown to be the result of charges on the membrane and of complexation with borate buffer ions. Investigation of "glutaraldehyde" solutions showed acid entities in, or formed in "glutaraldehyde" were responsible for EEO. Techniques for using "glutaraldehyde" which minimize EEO are described. PMID- 12652569 TI - Characterization of acridone dyes for use in four-decay detection in DNA sequencing. AB - Four acridone dyes and dye-labeled primers were characterized for use in four decay DNA sequencing. In the four-decay scheme, fluorescence lifetime replaces spectral ("color") selectivity for distinguishing between four base-specific labels in a single-lane capillary electrophoresis (CE) separation of the DNA fragments. Prior to the introduction of the acridone dyes, a major obstacle to four-decay detection was the lack of four suitable dyes with resolvable lifetimes. The four acridone dyes, whether free in solution or tethered to DNA primer, exhibit significant differences among their lifetimes and are well-suited to use together in four-decay sequencing. The lifetimes of the four dye-labeled DNA primers that were sequentially injected and detected on-the-fly in a 2% POP6 sequencing gel were 4, 6, 11 and 14 ns. A 405 nm violet laser diode provides optimal excitation of the four dyes. PMID- 12652570 TI - Efficient mutation detection in MEN1 gene using a combination of single-strand conformation polymorphism (MDGA) and heteroduplex analysis. AB - For facilitated genotypic analysis of multiple endocrine neoplasia type 1 (MEN1), a familial syndrome associated with tumors of the parathyroid and neuroendocrine tissues, we developed two screening methods, heteroduplex mutation assay (HMA) and mutation detection gel analysis (MDGA), both based on electrophoretic discrimination of polymerase chain reaction (PCR) products, to detect the mutations. Forty-three genomic DNA samples were used for the evaluation of these techniques. The whole coding region of MEN1 was PCR-amplified with fluorescent primers and then denatured/renatured before electrophoresis on an automated sequencer. 100% of the mutations were detected, subsequently confirmed and identified by sequencing. "Negative" samples were used to evaluate the specificity and reproducibility of the two techniques. The combination of the two methods allows high throughput cost-effective mutation screening which is less laborious than systematic sequencing of the whole coding region of MEN1. Together, these methods provide an efficient screen for MEN1 mutations. PMID- 12652571 TI - Microchannel wall coatings for protein separations by capillary and chip electrophoresis. AB - The necessity for microchannel wall coatings in capillary and chip-based electrophoretic analysis of biomolecules is well understood. The regulation or elimination of EOF and the prevention of analyte adsorption is essential for the rapid, efficient separation of proteins and DNA within microchannels. Microchannel wall coatings and other wall modifications are especially critical for protein separations, both in fused-silica capillaries, and in glass or polymeric microfluidic devices. In this review, we present a discussion of recent advances in microchannel wall coatings of three major classes--covalently linked polymeric coatings, physically adsorbed polymeric coatings, and small molecule additives. We also briefly review modifications useful for polymeric microfluidic devices. Within each category of wall coatings, we discuss those used to eliminate EOF, to tune EOF, to prevent analyte adsorption, or to perform multiple functions. The knowledgeable application of the most promising recent developments in this area will allow for the separation of complex protein mixtures and for the development of novel microchannel wall modifications. PMID- 12652572 TI - A DNA sieving matrix with thermally tunable mesh size. AB - We present a "proof-of-concept" study showing that a blend of thermo-responsive and nonthermo-responsive polymers can be used to create a DNA sieving matrix with a thermally tunable mesh size, or "dynamic porosity". Various blends of two well studied sieving polymers for CE, including hydroxypropylcellulose (HPC), a thermo responsive polymer, and hydroxyethylcellulose (HEC), a nonthermo-responsive polymer, were used to separate a double-stranded DNA restriction digest (Phi X174 HaeIII). HPC exhibits a volume-phase transition in aqueous solution which results in a collapse in polymer coil volume at approximately 39 degrees C. Utilizing a blend of HPC and HEC in a ratio of 1:5 by weight, we investigated the effects of changing mesh size on DNA separation, as controlled by temperature. High resolution DNA separations were obtained with the blended matrix at temperatures ranging from 25 degrees C to 38 degrees C. We evaluated changes in the selectivity of DNA separation with increasing temperature for certain pairs of small and large fragments. A pure HEC (nonthermo-responsive) matrix was used over the same temperature range as a negative control. In the blended matrix, we observe a maximum in selectivity at approximately 31 degrees C for small DNA, while a significant increase in the selectivity of large-DNA separation occurs at approximately 36 degrees C as the polymer mesh "opens". We also demonstrate, through a temperature ramping experiment, that this matrix can be utilized to obtain high-resolution separation of both small and large DNA fragments simultaneously in a single CE run. Blended polymer matrices with "dynamic porosity" have the potential to provide enhanced genomic analysis by capillary array or microchip electrophoresis in microfluidic devices with advanced temperature control. PMID- 12652573 TI - Direct identification of all oncogenic mutants in KRAS exon 1 by cycling temperature capillary electrophoresis. AB - Over the past few decades, advances in genetics and molecular biology have revolutionized our understanding of cancer initiation and progression. Molecular progression models outlining genetic events have been developed for many solid tumors, including colon cancer. Previous reports in the literature have shown a relationship between different KRAS mutations and prognosis and response to medical treatment in colon cancer patients. Furthermore, the presence of a mutated KRAS has been correlated with different clinicopathological variables including age and gender of patients and tumor location. To our knowledge, few institutions screen for KRAS mutations on regular basis in colon cancer patients despite such evidence that knowledge of KRAS exon 1 status is informative. Here, we report on a mutation analysis method adapted to a 96-capillary electrophoresis instrument that allows identification of all 12 oncogenic mutations in KRAS exon 1 under denaturing conditions. To determine the optimal parameters, a series of DNA constructs generated by site-directed mutagenesis was analyzed and the migration times of all mutant peaks were measured. A classification tree was then made based on the differences in migration time between the mutants and an internal standard. A randomized series of 500 samples constructed with mutagenesis as well as 60 blind samples from sporadic colon carcinomas was analyzed to test the method. No wild-type samples were scored as mutants and all mutants were correctly identified. Post polymerase chain reaction (PCR) analysis time of 96 samples was performed within 40 min. PMID- 12652574 TI - Capillary electrophoresis and fluorescence studies on molecular beacon-based variable length oligonucleotide target discrimination. AB - Molecular beacons (MBs) are oligonucleotide probes having a compact hairpin structure, with a fluorophore attached to one end and a quencher molecule attached to the other end. In its native state, the fluorophore is quenched by virtue of its proximity to the quencher molecule. Upon hybridization with its complementary oligonucleotide target, fluorescence is elicited due to a conformational change that results in separation of the fluorophore and quencher molecule. The present study describes the hybridization interaction of an MB to various complementary target sequences. The effects of temperature and length of complementary target sequences on hybridization were investigated using capillary electrophoresis and solution-based fluorescence techniques. Hybridization efficiency was dependent on the ability of the target sequences to destabilize the stem region by binding directly to the stem region. Optimal hybridization occurred between 40 and 50 degrees C for all targets tested, with the true target forming a more stable hybrid complex. PMID- 12652575 TI - Multiplexed DNA sizing by capillary electrophoresis using entangled polymer solutions and diode array detection. AB - We developed a method for the analysis of multiplexed double-stranded DNA (dsDNA) samples complexed to various intercalating dyes using entangled polymer solution. A commercial single-column capillary electrophoresis (CE) instrument with diode array detection was used for multiplexed detection of DNA samples by addition of intercalating fluorescent molecules. A Phi X174HinfI and a pGEM DNA ladder (1 mg/mL) were used for the electrophoretic separation of dsDNA fragments ranging in size from 24 to 726 and 36 to 2645 bp, respectively. The results suggested that simultaneous electrophoretic separation of different DNA ladders multiplexed with different dyes could be performed in the same capillary yielding fast DNA sizing separations. CE analysis, which is often overpowered by slab gel in sample throughput, could now overcome this disadvantage by allowing multiplexed sample analysis in a fraction of the time needed for slab gel analysis. The separation efficiency of stained DNA molecules with both dyes were dramatically improved with buffers containing a large cation such as tetrapentylammonium ion (Npe(4) (+)) as the only cation in the buffer. PMID- 12652576 TI - Micropreparative capillary gel electrophoresis of DNA: rapid expressed sequence tag library construction. AB - A capillary gel electrophoresis based automated DNA fraction collection technique was developed to support a novel DNA fragment-pooling strategy for expressed sequence tag (EST) library construction. The cDNA population is first cleaved by BsaJ I and EcoR I restriction enzymes, and then subpooled by selective ligation with specific adapters followed by polymerase chain reaction (PCR) amplification and labeling. Combination of this cDNA fingerprinting method with high-resolution capillary gel electrophoresis separation and precise fractionation of individual cDNA transcript representatives avoids redundant fragment selection and concomitant repetitive sequencing of abundant transcripts. Using a computer controlled capillary electrophoresis device the transcript representatives were separated by their size and fractions were automatically collected in every 30 s into 96-well plates. The high resolving power of the sieving matrix ensured sequencing grade separation of the DNA fragments (i.e., single-base resolution) and successful fraction collection. Performance and precision of the fraction collection procedure was validated by PCR amplification of the collected DNA fragments followed by capillary electrophoresis analysis for size and purity verification. The collected and PCR-amplified transcript representatives, ranging up to several hundred base pairs, were then sequenced to create an EST library. PMID- 12652577 TI - DNA fragment analysis by an affordable multiple-channel capillary electrophoresis system. AB - We are demonstrating a cost-effective multichannel capillary electrophoresis system for a high-efficiency double-stranded DNA (dsDNA) fragments analysis. This bench-type high-performance DNA analysis (HDA) system uses fluorescence-type detection with inexpensive solid-state light sources and nonmoving integrated emission collection micro-optics. DNA samples are analyzed simultaneously by using a multiple usage and disposable multicapillary cartridge, which contains integrated capillary channels, optical fibers and an integrated sieving gel reservoir. Using commercially available dsDNA size markers as indicators, the HDA system provides high resolving power in 7 min separations. The system can hold a total of 192 samples in two 96-well polymerase chain reaction (PCR) plates, which can be automatically analyzed within 2.5 h. This affordable system can be used in laboratories to replace slab gel electrophoresis for routine and high-throughput dsDNA analysis. PMID- 12652578 TI - Transcription factor binding study by capillary zone electrophoretic mobility shift assay. AB - Regulation of gene expression through interaction of proteins with specific DNA sequences is a central issue in functional genomics. Capillary electrophoretic mobility shift assay is an efficient novel method for the investigation of sequence specific protein-DNA interactions, allowing rapid and sensitive quantification of the complex formation. In this paper, we present a pilot study on capillary zone electrophoretic mobility shift assay (CZEMSA) to investigate the interaction between the transcription factors of HeLa nuclear extract and Sp1 specific fluorescein-labeled oligonucleotide, using the unlabeled probe as competitor. The mobility shift assay was accomplished by CZE in coated capillaries without polymeric buffer additives. Specificity of the DNA protein complex formation was verified by competition experiments, as well as by supershift assay with an anti-Sp1 antibody. The applied electric field strength did not affect the stability of DNA-protein complex during the electrophoretic analysis, allowing rapid identification and quantification of the protein DNA interaction. A practical application to study the interaction between Oryza sativa MADS-box transcription factor 4 (OsMADS4) and its consensus sequence is also reported. PMID- 12652579 TI - High-throughput screening of kinase inhibitors by multiplex capillary electrophoresis with UV absorption detection. AB - Protein kinases play a major role in the transformation of cells and are often used as molecular targets for the new generation of anticancer drugs. We present a novel technique for high-throughput screening of inhibitors of protein kinases. The technique involves the use of multiplexed capillary electrophoresis (CE) for the rapid separation of the peptides, phosphopeptides, and various inhibitors. By means of UV detection, diversified peptides with native amino acid sequences and their phosphorylated counterparts can be directly analyzed without the need for radioactive or fluorescence labeling. The effects of different inhibitors and their IC(50) value were determined using three different situations involving the use of a single purified kinase, two purified kinases, and crude cell extracts, respectively. The results suggest that multiplexed CE/UV may prove to be a straightforward and general approach for high-throughput screening of compound libraries to find potent and selective inhibitors of the various protein kinases. PMID- 12652581 TI - Capillary electrophoresis of proteins in dextran-coated columns. AB - A simple coating technique by using uncross-linked dextran has been developed for fused-silica capillaries to be used in capillary electrophoresis of basic proteins. The capillaries were first silanized with a heterobifunctional silane (gamma-aminopropyltriethoxylsilane), which served as a coupling agent between the capillary inner wall and the polysaccharide coating. Dextran of high molecular mass (about 70 kDa) was activated with 1,1'-carbonyldiimidazole. Then the activated dextran was coupled to the primary amino groups that were anchored onto the inner wall of the silanized capillaries. The residual reactive groups on the dextran were further substituted by neutral functions in a coupling reaction with excess ethanolamine. By using dimethyl sulfoxide (DMSO) rather than aqueous buffer as the reaction medium, the extent of substitution was improved by minimizing the residual reactive groups at the surface. Since they are ionogenic, the electrosmotic flow in the system is relatively low. The chemically bound dextran coating showed good reproducibility and stability. In electrophoretic experiments basic proteins were separated with high efficiency by use of the dextran-coated fused-silica capillary columns. The main advantage of the method described here is that both polysaccharide activation and amine-coupling reactions were carried out under mild conditions at room temperature without catalysts. For this reason, the method is recommended to coat the inner wall of microfluidic separation channels which would not tolerate a harsh treatment. PMID- 12652580 TI - High-throughput analysis of telomerase by capillary electrophoresis. AB - The enzyme telomerase is expressed in (85-90)% of all human cancers, but not in normal, non-stem cell somatic tissues. Clinical assays for telomerase in easily obtained body fluids would have great utility as noninvasive, cost-effective methods for the early detection of cancer. The most commonly used method for the detection and quantification of telomerase enzyme activity is the polymerase chain reaction (PCR)-based assay known as the telomerase repeat amplification protocol or TRAP assay. Most of the TRAP assay systems use a slab-gel based electrophoresis system to size and quantify the PCR-amplified extension products. We are developing high-throughput capillary electrophoresis (CE) methods for the analysis of TRAP/PCR products. The TRAP assay was conducted on lysates of the human lung cancer cell line A-549 in reactions containing 5-100 cells. TRAP/PCR products were generated using a fluorescent 4,7,2'4'5'7',-hexachloro-6 carboxyfluorescein(HEX)-labeled TS primer and analyzed on the Applied Biosystems Model 310 CE system using POP4 polymer. After analysis with GeneScan and Genotyper software, the total peak areas of the TRAP ladder extension products were computed using Microsoft Excel. Results were compared with unlabeled TRAP/PCR products analyzed on the Bio-Rad BioFocus 3000 CE system using 6% high molecular weight polyvinylpyrrolidone (HMW PVP) polymer and SYBR Green I dye. Both CE systems were able to resolve the TRAP ladder products with high reproducibility and sensitivity (5-15 cells). With the appropriate robotic sample handling system, these CE methods would enable performing the telomerase TRAP assay with increased sensitivity, reproducibility and automation over slab-gel methods. PMID- 12652582 TI - Mechanism of action of quaternary diamino quenchers in capillary zone electrophoresis. AB - The synthesis of two novel amino compounds, able to quench and reverse the electroosmotic low (EOF) in capillary electrophoresis is here reported. These chemicals are derivatives of two previously described quaternarized piperazines, 1-(4-iodobutyl)-1,4-dimethylpiperazin-1-ium iodide (M1C4I) and 1-(4-iodobutyl)4 aza-1-azoniabicyclo[2.2.2] octane iodide (M7C4I), believed to bind covalently to the silica surface via alkylation of ionized silanols through their terminal, reactive iodine. The novel compounds, although unable to form a covalent bond, due to lack of the o-butyl iodine, are found to be very efficient in suppressing protein interaction with the wall and reversing the EOF. On the basis of their behavior in solution, the minimal structural motifs for strong binding of amino compounds to the silica wall have been derived and are thought to be: (i) the presence of two quaternary nitrogens in the molecule; (ii) a correct distance between said charged nitrogens, represented by a butyl chain (C(4)); (iii) an hydrophobic "decoration" of the molecules, consisting on a high CH(2)/N ratio (in the present case 8:1). PMID- 12652583 TI - A family of single-isomer, sulfated gamma-cyclodextrin chiral resolving agents for capillary electrophoresis: octa(6-O-sulfo)-gamma-cyclodextrin. AB - The second member of the single-isomer, sulfated gamma-cyclodextrin family, the sodium salt of octa(6-O-sulfo)-gamma-cyclodextrin (OS) has been synthesized, characterized and used to separate the enantiomers of nonelectrolyte, acidic, basic, and ampholytic analytes by capillary electrophoresis in acidic aqueous background electrolytes. The anionic effective mobilities of the nonelectrolyte and anionic analytes increased with increasing concentration of OS. The effective mobilities of strongly complexing cationic analytes became anionic with very low OS concentrations and passed local anionic effective mobility maxima as the OS concentration, and along with it, the ionic strength, of the background electrolyte increased. The effective mobilities of the weakly binding cationic analytes became only slightly anionic at high OS concentration values and did not show the local anionic effective mobility maxima. For nonelectrolyte analytes, separation selectivities decreased with increasing OS concentration. For cationic analytes, separation selectivities were highest where the effective mobilities of the less mobile enantiomers approached zero. OS proved to be a broadly applicable chiral resolving agent. PMID- 12652584 TI - Characterization of pharmaceutical drugs by a modified nonaqueous capillary electrophoresis--mass spectrometry method. AB - A simple method for the separation and characterization of a group of nine basic compounds, comprising seven tricyclic antidepressant and two bronchodilator drugs, by nonaqueous capillary electrophoresis (NACE) employing ultraviolet and mass spectrometry detection is described. After optimization of the electrophoresis separation conditions, including the compositions of the electrolyte and the organic solvent, a reliable separation of all nine basic analytes was achieved in 80 mM ammonium formate dissolved in a methanol acetonitrite (80:20 v/v) mixture, having an apparent pH of 8.7. The volatile nonaqueous electrolyte system used with a normal electroosmotic flow polarity also provided an optimal separation condition for the characterization of the analytes by mass spectrometry. When results were compared with reversed-phase gradient and isocratic high-performance liquid chromatography (HPLC) methods, the NACE method provided greater efficiency, achieving baseline resolution for all nine basic compounds in less than 30 min. The NACE method is suitable for use as a routine procedure for the rapid separation and characterization of basic compounds and is a viable alternative to HPLC for the separation of a wide range of pharmaceutical drugs. PMID- 12652585 TI - A versatile microfabricated platform for electrophoresis of double- and single stranded DNA. AB - We demonstrate a versatile microfabricated electrophoresis platform, incorporating arrays of integrated on-chip electrodes, heaters, and temperature sensors. This design allows a range of different sieving gels to be used within the same device to perform separations involving both single- and double-stranded DNA over distances on the order of 1 cm. We use this device to compare linear and cross-linked polyacrylamide, agarose, and thermo-reversible Pluronic-F127 gels on the basis of gel casting ease, reusability, and overall separation performance using a 100 base pair double-stranded DNA ladder as a standard sample. While cross-linked polyacrylamide matrices provide consistently high-quality separations in our system over a wide range of DNA fragment sizes, Pluronic gels also offer compelling advantages in terms of the ability to remove and reload the gel. Agarose gels offer good separation performance, however, additional care must be exercised to ensure consistent gel properties as a consequence of the need for elevated gel loading temperatures. We also demonstrate the use of denaturing cross-linked polyacrylamide gels at concentrations up to 19% to separate single-stranded DNA fragments ranging in size from 18 to 400 bases in length. Primers differing by 4 bases at a read length of 30 bases can be separated with a resolution of 0.9-1.0 in under 20 min. This level of performance is sufficient to conduct a variety of genotyping assays including the rapid detection of single nucleotide polymorphisms (SNPs) in a microfabricated platform. The ability to use a single microelectrophoresis system to satisfy a wide range of separation applications offers molecular biologists an unprecedented level of flexibility in a portable and inexpensive format. PMID- 12652586 TI - Single-nucleotide polymorphism analysis by allele-specific extension of fluorescently labeled nucleotides in a microfluidic flow-through device. AB - We describe a microfluidic approach for allele-specific extension of fluorescently labeled nucleotides for scoring of single-nucleotide polymorphism (SNP). The method takes advantage of the fact that the reaction kinetics differs between matched and mismatched configurations of allele-specific primers hybridized to DNA template. A microfluidic flow-through device for biochemical reactions on beads was used to take advantage of the reaction kinetics to increase the sequence specificity of the DNA polymerase, discriminating mismatched configurations from matched. The volume of the reaction chamber was 12.5 nL. All three possible variants of an SNP site at codon 72 of the p53 gene were scored using our approach. This work demonstrates the possibility of scoring SNP by allele-specific extension of fluorescently labeled nucleotides in a microfluidic flow-through device. The sensitive detection system and easy microfabrication of the microfluidic device enable further miniaturization and production of an array format of microfluidic devices for high-throughput SNP analysis. PMID- 12652587 TI - Comparison of capillary zone electrophoresis performance of powder-blasted and hydrogen fluoride-etched microchannels in glass. AB - The applicability of glass chips with powder-blasted microchannels for electrophoretic separations was examined, and the performance was compared to microchannels etched with hydrogen fluoride (HF), using bicarbonate buffer and rhodamine B and fluorescein as model compounds. The measured electroosmotic mobilities in all chips were comparable, with values of ca. 7 x 10(-4) cm(2) V( 1)s(-1). The effect of electrical field strength and detection length on the separation efficiency was monitored. It was found that the main source of dispersion is of the Taylor-Aris type, which was discussed in relation to channel roughness differences. Although in powder-blasted channels with a separation length of 8.20 cm, 7-9 times lower plate numbers were obtained than in a HF etched channel with similar dimensions, successful separation of five fluorescein isothiocyanate (FITC)-labeled amino acids was obtained on a powder-blasted chip within 80 s. Efficiencies of up to 360 000 plates/m were demonstrated on this chip, when a higher buffer concentration was used at a field strength of 664 V/cm. It can be concluded that powder-blasted microchannel chips, although they have a lower separation efficiency compared to HF-etched chips, perform well enough for many applications. Powder blasting can therefore be considered a low cost and efficient alternative to HF etching, in particular because of the possibility to fabricate access holes through the glass with the same process. PMID- 12652588 TI - Practical integration of polymerase chain reaction amplification and electrophoretic analysis in microfluidic devices for genetic analysis. AB - An integrated system of a silicon-based microfabricated polymerase chain reaction (microPCR) chamber and microfabricated electrophoretic glass chips have been developed. The PCR chamber was made of silicon and had aluminum heaters and temperature sensors integrated on the glass anodically bonded cover. Temperature uniformity in the reaction chamber was +/-0.3 degrees C using an improved novel "joint-heating" scheme. Thermal cycling was digitally controlled with a temperature accuracy of +/- 0.2 degrees C. Small operating volumes together with high thermal conductivity of silicon made the device well suited to rapid cycling; 16 s/cycle were demonstrated. For analysis of the PCR products, the chamber output was transferred to the glass microchip by pressure. Analysis time of PCR amplified genomic DNA was obtained in the microchip in less than 180 s. The analysis procedure employed was reproducible, simple and practical by using viscous sieving solutions of hydroxypropylmethylcellulose and dynamically coated microchip channels with poly(vinylpyrrolidone). DNA fragments that differ in size by 18 base pairs (bp) were resolved. Analysis of genomic male and female amplified DNA by microPCR was achieved in microchip, and application of the integrated microPCR-microchip for the identification of bird sex was tested. Genomic DNA samples from several bird species such as pigeon and chicken were analyzed. Hence, the system could be used as well to determine the sex of avian species. PMID- 12652589 TI - An interface chip connection between capillary electrophoresis and thermal lens microscope. AB - A thermal lens microscope (TLM) detection of capillary electrophoresis (CE) utilizing microchip technology was developed. Fused-silica capillaries with an inner diameter of 50 microm were directly connected to a microchannel in a microchip. The detection limit by TLM was estimated as 2.8 x 10(-7) absorbance by measuring pure water. The detection limit of derivatized amino acids determined by CE-TLM was estimated as 2.4 x 10(-8) M, which was 100 times lower than that of conventional absorbance detection. PMID- 12652590 TI - Low-voltage electroosmosis pump for stand-alone microfluidics devices. AB - Two types of low-voltage electroosmosis pumps were developed using microfabrication technology for usage in handy or stand-alone applications of the micrototal analysis systems (micro-TAS) and the lab-on-a-chip. This was done by making a thin (< 1 microm) region in the flow path and by only applying voltages near this thin region using electrodes inserted into the flow path. The inserted electrodes must be free from bubble formation and be gas-tight in order to avoid pressure leakage. For these electrodes, Ag/AgCl or a gel salt bridge was used. For patterning the gel on the chip, a hydrophilic photopolymerization gel and a photolithographic technique were optimized for producing a gel with higher electric conductivity and higher mechanical strength. For high flow rate application, wide (33.2 mm) and thin (400 nm) pumping channels were compacted into a 1 mm x 6 mm area by folding. This pump achieves an 800 Pa static pressure and a flow of 415 nL/min at 10 V. For high-pressure application, a pump was designed with the thin and thick regions in series and positive and negative electrodes were inserted between them alternatively. This pump could increase the pumping pressure without increasing the supply voltage. A pump with 10-stage connections generated a pressure of 25 kPa at 10 V. PMID- 12652591 TI - Dynamic analyte introduction and focusing in plastic microfluidic devices for proteomic analysis. AB - Isoelectric focusing (IEF) separations, in general, involve the use of the entire channel filled with a solution mixture containing protein/peptide analytes and carrier ampholytes for the creation of a pH gradient. Thus, the preparative capabilities of IEF are inherently greater than most microfluidics-based electrokinetic separation techniques. To further increase sample loading and therefore the concentrations of focused analytes, a dynamic approach, which is based on electrokinetic injection of proteins/peptides from solution reservoirs, is demonstrated in this study. The proteins/peptides continuously migrate into the plastic microchannel and encounter a pH gradient established by carrier ampholytes originally present in the channel for focusing and separation. Dynamic sample introduction and analyte focusing in plastic microfluidic devices can be directly controlled by various electrokinetic conditions, including the injection time and the applied electric field strength. Differences in the sample loading are contributed by electrokinetic injection bias and are affected by the individual analyte's electrophoretic mobility. Under the influence of 30 min electrokinetic injection at constant electric field strength of 500 V/cm, the sample loading is enhanced by approximately 10-100 fold in comparison with conventional IEF. PMID- 12652592 TI - Integration of on-column immobilized enzyme reactor in microchip electrophoresis. AB - A simple method integrating an immobilized enzyme reactor into a microchip electrophoresis device was developed. The enzyme immobilization into a microchip was performed by spotting and drying a drop of dissolved nitrocellulose (NC) on a glass substrate, and adsorbing enzyme on the reconstituted NC membrane. This enzyme-immobilized glass plate was assembled with a polydimethylsiloxane substrate on which the separation channel was fabricated. The advantage of this method is the ability to easily change the position and size of the reactor within the microchip electrophoresis device. A beta-galactosidase reaction was demonstrated with fluorescein di-beta-D-galactopyranoside using this integrated on-column enzyme reactor. A successful electrophoretic separation of its hydrolysis products, i.e., fluorescein mono-beta-D-galactopyranoside (FMG) and fluorescein, was achieved. Enzyme kinetics and inhibition of the beta galactosidase using FMG and 2-phenylethyl beta-D-thiogalactoside, respectively, were also studied with microchip electrophoresis. PMID- 12652593 TI - Extraction of proteins from plant tissues for two-dimensional electrophoresis analysis. AB - To increase the number of proteins detectable by two-dimensional electrophoresis (2-DE) in plants, we present a new procedure for extracting total proteins from plant tissue. This method avoids any loss of proteins in the course of sample preparation and results in two different fractions, one comprising mainly the cytoplasmatic proteins, the other one containing predominantly structure bond proteins. 2-DE patterns obtained from these two fractions show that the total number of different protein spots detected exceeds the degree of resolution commonly reported for plant proteins threefold. PMID- 12652594 TI - Spot overlapping in two-dimensional polyacrylamide gel electrophoresis maps: relevance to proteomics. AB - Proteomics requires a large-scale, simultaneous separation of proteins from a mixture, assessment of the relative abundance of these molecules, and identification and characterization of each component. In 2-D PAGE separations, the best method of choice for protein analysis, separation of all the proteins present in the sample is still far to be achieved and comigrating proteins in the same spot are in general present. A statistical estimation of the degree of spot overlapping present in a 2-D PAGE separation is here described: for different conditions of spot overcrowding in the map, the degree of overlapping can be quantified in terms of purity degree of each spot or percentage of proteins that will appear in the map as a single spot. A computer simulation approach is described: it is based on the protein separation pattern present in the experimental maps. The results thus obtained are compared to a theoretical model (statistical degree of peak overlapping model) based on random spot position. The described procedures were applied to an experimental reference map of human plasma. The severity of spot overlapping in 2-D PAGE maps is estimated and the influence of different experimental conditions (strip dimension, detector system performance, pI range) is discussed. These informations are useful to quantitatively estimate the degree of error associated with identification and quantitation of each protein and to set-up experimental conditions which will increase resolution and greatly decrease the probability of spot overlapping. PMID- 12652595 TI - A new integrated statistical approach to the diagnostic use of two-dimensional maps. AB - Two-dimensional (2-D) electrophoresis is a very useful technique for the analysis of proteins in biological tissues. The complexity of the 2-D maps obtained causes many difficulties in the comparison of different samples. A new method is proposed for comparing different 2-D maps, based on five steps: (i) the digitalisation of the image; (ii) the transformation of the digitalised map in a fuzzy entity, in order to consider the variability of the 2-D electrophoretic separation; (iii) the calculation of a similarity index for each pair of maps; (iv) the analysis by multidimensional scaling of the previously obtained similarity matrix; (v) the analysis by classification or cluster analysis techniques of the resulting map co-ordinates. The method adopted was first tested on some simulated samples in order to evaluate its sensitivity to small changes in the spots position and size. The optimal setting of the method parameters was also investigated. Finally, the method was successfully applied to a series of real samples corresponding to the electrophoretic bidimensional analysis of sera from normal and nicotine-treated rats. Multidimensional scaling allowed the separation of the two classes of samples without any misclassification. PMID- 12652597 TI - Proteome analysis using isoelectric focusing in immobilized pH gradient gels followed by mass spectrometry. AB - Over the past several years, a large effort has been focused on improvements of two-dimensional (2-D) gel electrophoresis-based proteomics technology, and on development of novel approaches for proteome analysis. Here, we describe the application of an alternative strategy for the analysis of complex proteomes. The strategy combines isoelectric focusing in immobilized pH gradient strips (in-gel IEF), mass spectrometry (MS), and bioinformatics. A protein mixture is separated by in-gel IEF, and the entire strip is cut into a set of gel sections. Proteins in each gel section are digested with trypsin, and the tryptic peptides are subjected to liquid chromatography-nanoelectrospray-quadrupole ion-trap tandem mass spectrometry (LC-ESI-MS/MS). The LC-ESI-MS/MS data are used to identify the proteins through searches of a protein sequence database. Using this in-gel IEF LC-MS/MS strategy, we have identified 127 proteins from a human pituitary. This study demonstrates the potential of the in-gel IEF-LC-MS/MS approach for analyses of complex mammalian proteomes. PMID- 12652596 TI - Identification of nuclear proteins of small cell lung cancer cell line H82: An improved procedure for the analysis of silver-stained proteins. AB - An efficient method for digestion and extraction of proteolytic peptides from silver-stained proteins was applied to the characterization of nuclear proteins from the small cell lung cancer H82 (ATCC HTB 175) cell line previously separated by high-resolution large format two-dimensional gel electrophoresis. From 68 spots, evenly distributed on the gel area and representing a wide range of spot intensities, 63 (92%) were successfully identified by matrix-assisted laser desorption/ionization (MALDI) or electrospray ionozation-mass spectrometry (ESI MS). In five cases where the identification was not possible, the presence of an intense background apparently due to the leakage of polymers from the microtubes or other plastics, was detected. Extensive analysis of peptide sequences by ESI MS/MS experiments allowed the identification of post-translational modifications, such as acetylation, phosphorylation, deamidation of asparagine residues and the presence of isoaspartic acid. A new protein variant not reported in sequence databases was also detected. PMID- 12652598 TI - The mitochondrial proteins of the neuroblastoma cell line IMR-32. AB - Mitochondrial proteins exert important functions in biological pathways, particularly they are involved in apoptotic processes. We applied proteomics technologies to analyze the mitochondrial proteins of the neuroblastoma cell line IMR-32, which is often used in apoptosis studies. The proteins were analyzed by two-dimensional (2-D) electrophoresis followed by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS). 185 different gene products were identified, of which approximately 55% were enzymes with a broad spectrum of catalytic activities. Sixteen proteins were detected only in this preparation, the others have been detected in two or more protein samples analyzed by MS in our laboratory. The 16 unique gene products were represented by one spot each, whereas most of the frequently detected proteins were represented by multiple spots. In average, approximately 5-10 spots corresponded to one gene product. For two thirds of the proteins identified, an annotation exists in the SWISS-PROT database about their subcellular location. They are mainly described as mitochondrial, 8 as endoplasmic reticulum, 3 as peroxisomal and only 12 low abundance proteins are described as cytosolic proteins. The list includes about 30 unknown, hypothetical or poorly described gene products. Some of them are represented by strong spots and the present study shows that they are indeed expressed and are localized in the mitochondria. PMID- 12652599 TI - Activation of platelet-derived growth factor pathway in human asthmatic pulmonary derived mesenchymal cells. AB - Cell cultures of mesenchymal type were obtained from biopsies taken after bronchoscopy from patients with asthma. It was possible to achieve outgrowth of fibroblast-like cells from these lung biopsies, which stained for alpha-smooth actin indicating that they were of myofibroblast type. Morphologically, two types of myofibroblasts could be observed: one intermediate form with more stretched cell shape and lamellipodia protrusions, and one more differentiated compact form of myofibroblast. The intermediate form was the most dominant type in these patients, indicating an active ongoing remodelling process. Further studies showed that platelet-derived growth factor (PDGF) might be the factor that stimulates the formation of the intermediate type of myofibroblasts, since it enhance migration of normal human lung fibroblasts 4-fold compared to control through an induced formation of stress fibers and lamellipodia protrusions. Additionally, intracellular signalling pathways involved in migration, such as RhoA and MAPkinase were stimulated 1.5-fold and 3.5-fold, respectively. By using two-dimensional (2-D) gel electrophoresis and protein identification by peptide mass finger printing matrix assisted laser desporption/ionization - time of flight - mass spectrometry (MALDI-TOF-MS) it was possible to confirm that PDGF affected the synthesis of proteins involved in the remodelling process, such as collagen VI and post-translational forms thereof. PDGF also stimulated the production of FK506 binding protein of 65 kDa, a protein involved in smooth muscle differentiation, and proteins involved in the rearrangement of the cytoskeleton connected to migration such as the actin related protein ARP3, the T complex protein and the heat shock protein 60. We demonstrate that PDGF has a potential pathological role in asthma and formation of subepithelial fibrosis by inducing changes in the proteome. PMID- 12652600 TI - Two-dimensional protein map of human vastus lateralis muscle. AB - Protein samples of human vastus lateralis muscle were analyzed by two-dimensional gel electrophoresis, using immobilized pH gradients encompassing several pH regions in the first dimension and sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the second dimension. More than 500 protein spots on each gel were detected by silver staining, of which 150 were excised, digested in-gel with trypsin and characterized by matrix assisted laser desorptioin/ionization-mass spectrometry and tandem electrospray mass spectrometry. Using these techniques, 124 spots including contractile proteins and metabolic enzymes were identified. This database should provide a valuable resource for the definition of the functional properties of muscle fibres and for assessment of the influence of e.g., hypoxia, ischaemia and ageing on protein level. PMID- 12652601 TI - Proteomic analysis of immunostained, laser-capture microdissected brain samples. AB - Proteomic analysis is often performed on homogenized preparations of whole tissues, which does not provide any information about relevant biochemical changes in specific cell types. Laser-capture microdissection (LCM) is a technique that is precise enough to dissect single cells within a tissue section. Phenotypically defined cells of interest may be visualized by immunostaining prior to microdissection. Previously published immunostaining protocols adapted to LCM require the use of very high antibody titers and very short incubation times. This raises the concern that low-abundance antigens would not be detected and that antisera would be rapidly depleted. In addition, protein recovery from samples was not evaluated in most of these studies. Here, we describe an optimized immunostaining method based on immunofluorescence. By comparing two dimensional electrophoresis (2-DE) results obtained from immunostained LCM brain tissue samples to those obtained from unstained, manually dissected samples, we demonstrated that immunofluorescent staining gave comparable protein recovery and similar resolution of protein spots on 2-DE gels. Moreover, matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry analysis of selected spots from gels derived from control and immunostained LCM samples revealed that the immunostaining process had minimal effect on protein identification. LCM of immunofluorescently labeled tissue sections is a practical and powerful method to perform proteomic studies on specifically defined cell groups. PMID- 12652602 TI - Monitoring of protein profiles for the optimization of recombinant fermentation processes using public domain databases. AB - The expression of human superoxide dismutase in fed-batch fermentation of E. coli HMS174(DE3)(pET3ahSOD) was studied as model system. Due to the frequently used strong T7 promoter system a high metabolic load is exerted, which triggers stress response mechanisms and finally leads to the differentiation of the host cell. As a consequence, host cell metabolism is partly shifted from growth to survival accompanied by significant alterations of the protein pattern. In terms of process optimization two-dimensional electrophoresis deserves as a powerful tool to monitor these changes on protein level. For the analysis of samples derived from different states of recombinant protein production wide-range Immobiline Dry Strips pH 3-10 were used. In order to establish an efficient procedure for accelerated process optimization and to avoid costly and time-consuming analysis like mass spectrometry (MS), a database approach for the identification of significant changes of the protein pattern was evaluated. On average, 935 spots per gel were detected, whereby 50 are presumably stress-relevant. Out of these, 24 proteins could be identified by using the SWISS-2DPAGE database (www.expasy.ch/ch2d/). The identified proteins are involved in regulatory networks, energy metabolism, purine and pyrimidine nucleotide synthesis and translation. By this database approach, significant fluctuations of individual proteins in relation to recombinant protein production could be identified. Seven proteins show strong alterations (>100%) directly after induction and can therefore be stated as reliable marker proteins for the assessment of stress response. For distinctive interpretation of this highly specific information, a bioinformatic and statistic tool would be essential in order to perceive the role and contribution of individual proteins in stress response. PMID- 12652605 TI - Editors and conflict of interest. PMID- 12652606 TI - Situated clinical reasoning: distinguishing acute confusion from dementia in hospitalized older adults. AB - In this study a dimensional analysis approach was used to explore the clinical reasoning of nurses who care for hospitalized older adults to identify factors that might explain their failure to detect acute confusion and to distinguish it from dementia in this patient population. Data analysis yielded a grounded theory of situated clinical reasoning, which proposes that the ability of nurses to identify acute confusion varies widely. This variation can be attributed to the differences in nurses' philosophical perspectives on aging. According to this theory, three distinct perspectives are unwittingly embraced by nurses who care for older patients. These perspectives influence how nurses characterize aging and the aged and condition the ways in which they judge and ultimately deal with older adults in clinical situations. PMID- 12652608 TI - Screening for risk of rehospitalization from home care: use of the outcomes assessment information set and the probability of readmission instrument. AB - The purpose of this study was to evaluate the Outcomes Assessment Information Set (OASIS) compared with the Probability of Readmission (P(ra)) instrument for use in predicting rehospitalization during home care. Using logistic regression and receiver operating characteristic (ROC) curve analysis, the P(ra) instrument was found to be significantly better at predicting rehospitalization than the OASIS case mix weight, clinical, or service scores. The area under the curve (AUC) for the P(ra) was .686 compared with .549 for the OASIS case mix weight (p =.010). Similar results were found for the OASIS clinical and service scores. The AUC for the function score of >/=2 (.599) provided the closest approximation to the P(ra) (.686), and the difference between the two was not statistically significant (p =.120). The OASIS function score could be used to identify at-risk home care patients without having to also use the P(ra) instrument. PMID- 12652607 TI - Effects of stress management on PNI-based outcomes in persons with HIV disease. AB - A pretest-posttest, repeated-measures design was used to evaluate the effects of two stress management interventions on a battery of outcomes derived from a psychoneuroimmunological (PNI) framework. The effects of cognitive-behavioral relaxation training groups (CBSM) and social support groups (SSG) were compared with a WAIT-listed control group on the outcomes of psychosocial functioning, quality of life, neuroendocrine mediation, and somatic health. Participants were 148 individuals (119 men, 29 women), diagnosed with HIV disease; 112 (76%) completing the study groups. Using analysis of covariance, the CBSM group was found to have significantly higher postintervention emotional well-being and total quality-of-life scores than did either the SSG or WAIT groups. SSG participants had significantly lower social/family well-being scores immediately postintervention and lower social support scores after 6 months. The findings point to a pressing need for further, well-controlled research with these common intervention modalities. PMID- 12652609 TI - Teaching nursing research to undergraduates: a text analysis of instructors' intentions. AB - Reviews of teaching strategies for undergraduate nursing research have been organized according to (a) the type of learning to be achieved, such as learning by doing, or (b) the specific teaching strategy, such as a poster session. For this text analysis, a new tack was taken to reveal the intentions of undergraduate nursing research instructors for student learning. Giorgi's (1985) descriptive phenomenological method was used to analyze 77 narrative reports of instructors about research teaching strategies. Seven intentions were identified, including desensitizing negative perceptions about research and stimulating collaborative learning about research. The intentions were contrasted in scope and relevance to frameworks organized according to learning goals or teaching techniques. The relevance of the intentions was considered in relation to critical trends influencing nursing education. PMID- 12652610 TI - Psychometric characteristics of the breastfeeding self-efficacy scale: data from an Australian sample. AB - Many new mothers discontinue breastfeeding prematurely because of difficulties encountered rather than maternal choice. Research has shown that a significant predictor of breastfeeding duration is a mother's confidence in her ability to breastfeed. To measure breastfeeding confidence, the Breastfeeding Self-Efficacy Scale (BSES) was developed and psychometrically tested at 1 week postpartum. The purpose of this methodological study was to psychometrically test the BSES antenatally and at 1 week and 4 months postpartum in a sample of Australian women and to determine predictive validity. The psychometric assessment of the original BSES study was replicated, including internal consistency, principal components factor analysis, comparison of contrasted groups, and correlations with a similar construct. Support for predictive validity was demonstrated through positive correlations and significant mean differences between antenatal BSES scores and infant-feeding method at 1 week and 4 months postpartum. The BSES is now considered ready for both research and clinical use (a) to identify new mothers with low breastfeeding confidence who require additional assistance, (b) to assess breastfeeding behaviors and cognitions in order to have individualized confidence-building strategies, and (c) to assist in the evaluation of various nursing interventions. PMID- 12652611 TI - Toward a metasynthesis of qualitative findings on motherhood in HIV-positive women. AB - A qualitative metasynthesis of qualitative findings ought to be more than a mere summary of those findings. Yet the processes by which the interpretive innovation expected of qualitative metasynthesis projects can be achieved remain opaque. Several analytic devices for the metasynthesis of findings were clarified in the course of an ongoing methodological project involving 45 reports of qualitative studies of HIV-positive women. These devices include the creation of a taxonomy of findings, the explicit use of sustained comparisons, the translation of in vivo concepts, and the use of imported concepts. Any qualitative metasynthesis of findings constitutes an interpretation at least three times removed from the lives represented in them. Clarifying the analytic devices used to create such metasyntheses is essential to demonstrating that despite being far away from participants' lives, these interpretations remain close to them. PMID- 12652612 TI - Thoughts on "It's time to revalue nursing education research". PMID- 12652614 TI - Squamous carcinoma of the larynx and hypopharynx in children: An enigma and a dilemma. PMID- 12652615 TI - Treatment of Ewing sarcoma family of tumors: current status and outlook for the future. AB - BACKGROUND: The Ewing sarcoma family of tumors (ESFT) comprises a group of well characterized neoplasms with aggressive behavior. Despite significant progress with the use of intensive multiagent chemotherapy and local control measures, a significant proportion of patients die of disease progression. Chemotherapy dose intensification and autologous hematopoietic stem cell transplant (HSCT) have been explored by many institutions without obvious benefit in high-risk patients. Our current understanding in the biology and treatment of ESFT suggests that a more rational approach to the development of risk-adapted therapy should be undertaken. PROCEDURE: We performed a review of the most relevant data regarding the current status in the treatment of ESFT. The results of the major American and European cooperative groups were analyzed, including the treatment strategies used and the prognostic factors identified for both localized and metastatic ESFT. RESULTS: The intensification of alkylating agents and topoisomerase-II inhibitors is feasible and has resulted in some survival improvement for selected patients. This benefit seems to be restricted to patients with localized disease, and a proportion of survivors are at risk of developing treatment-related hematologic malignancies. Nevertheless, these advances have resulted in a re definition of prognostic factors, which may help to define risk groups based on tumor load parameters as well as biologic factors (type of fusion transcript and histologic response to chemotherapy). Patients with advanced metastatic disease may benefit from HSCT. New strategies such as immunotherapy and the use of biologic modifiers may have a role in the treatment of ESFT. CONCLUSIONS: Future treatment for ESFT should consider risk-adapted strategies and the inclusion of newer therapies such as biologic modifiers for the minimal residual disease. A modified risk-adapted therapy is proposed. PMID- 12652616 TI - Non-resectable congenital tumors with the ETV6-NTRK3 gene fusion are highly responsive to chemotherapy. AB - BACKGROUND: Recently, the ETV6-NTRK3 gene fusion has been identified in both infantile fibrosarcoma and cellular mesoblastic nephroma. For both these tumors standard curative treatment has been primarily surgical with wide local excision. This has frequently involved radical and even mutilating surgery. PROCEDURE: This report discusses three infants with congenital tumors, two congenital fibrosarcomas, and one atypical congenital mesoblastic nephroma, not easily amenable to surgical intervention. RESULTS: All three were treated with pre operative chemotherapy with excellent responses negating the need for amputation in two patients. In each patient, the ETV6-NTRK3 gene fusion was identified by reverse transcriptase-polymerase chain reaction (RT-PCR) in the tumor specimens. CONCLUSIONS: Our findings suggest that the ETV6-NTRK3 gene fusion may underlie the distinctive biological properties of these tumors and may also indicate tumor chemosensitivity. In this group of patients pre-operative chemotherapy may abrogate the need for morbid surgical procedures. PMID- 12652617 TI - Pharmacology of all-trans-retinoic acid in children with acute promyelocytic leukemia. AB - BACKGROUND: Due to severe side effects in virtually all children treated with a standard dose of 45 mg/m(2)/day all-trans-retinoic acid (ATRA) for acute promyelocytic leukemia (APL) the AML-BFM study group reduced the dosage to 25 mg/m(2)/day. For the lack of data on the use of ATRA at this dosage in children with APL, the study group further decided to evaluate the pharmacokinetics and metabolism of ATRA in children. PROCEDURE: Twenty-three pharmacokinetic and metabolic profiles of ATRA were studied in 14 children (aged 0.9-18.4 years) with APL. Eleven plasma samples were collected over a period of 8 hr and analyzed for ATRA and its metabolites by high-performance liquid chromatography. RESULTS: Peak plasma concentrations of ATRA were characterized by wide interpatient variability (range: 28.6-513.0 nM). Compared to adults the same metabolic pathways were observed in children. Even though peak plasma concentrations were in the lower range of those considered effective in vitro, ATRA side effects, notably neurotoxicity, still required dose reduction, treatment break, or drug withdrawal in eight patients. In this small number of patients, neurotoxicity could not be related to age or any specific level of ATRA or metabolites in the plasma. Plasma concentrations of vitamin A, however, were significantly higher in those patients, who developed signs of neurotoxicity (P = 0.03, Mann-Whitney Rank Sum test). CONCLUSIONS: Considering the low plasma concentrations and the persistence of toxicity in spite of dose reduction intermittent dosing schedules might be considered as an alternative to further dose reduction of ATRA in the treatment of APL especially in children, who might be at risk of ATRA-induced neurotoxicity. PMID- 12652618 TI - Expression of the lipogenic enzyme fatty acid synthase (FAS) as a predictor of poor outcome in nephroblastoma: an interinstitutional study. AB - BACKGROUND: Treatment of nephroblastoma (Wilms tumor) has presently achieved a greater than 80% cure rate. Pathologic stage and grade are considered the most reliable prognostic parameters, but other biologic factors are under study in order to improve patient stratification into risk groups. Correlation of elevated levels of the lipogenic enzyme fatty acid synthase (FAS) with aggressiveness of some cancers has drawn attention to this enzyme as a possible marker of poor prognosis. PROCEDURE: To determine the predictive strength of FAS expression in Wilms tumor (with particular emphasis on intermediate risk, i.e., non anaplastic tumors, the vast majority of nephroblastomas), we evaluated immunostaining expression in archival specimens from 94 neoplasms. The degree of expression was correlated with stage, grade, clinical course and administration of prenephrectomy chemotherapy. RESULTS: Expression of FAS increased in anaplastic tumors (P = 0.043) and higher stages (P = 0.029). FAS expression correlated with OS and DFS at both univariate and multivariate analysis. Comparable results were obtained when analyzing the intermediate risk population separately. Pretreatment resulted in an increased FAS expression, without reaching significance level (P = 0.059). CONCLUSIONS: Expression of FAS might be an independent prognostic factor, particularly for intermediate-risk patients. The blockade of fatty acid synthesis by inhibition of FAS enzymatic function by means of metabolic analogues might prove a novel target pathway for the treatment of nephroblastoma. PMID- 12652619 TI - Proton beam therapy in the management of central nervous system tumors in childhood: the preliminary experience of the Centre de Protontherapie d'Orsay. AB - BACKGROUND: The purpose of the study was to evaluate clinical results and complications of a combination of proton and photon irradiation administered to 17 children with selected central nervous system (CNS) tumors. PROCEDURE: Between July 1994 and September 2000, 17 children, aged from 5 to 17 years (median: 12 years) with intracranial benign (6 cases) or malignant (11 cases) tumors, were treated with photons (median dose: 40 Gy; 24-54) and protons (median dose: 20 CGE; 9-31) at the Centre de Protontherapie d'Orsay (CPO). RESULTS: Mean follow-up was 27 months (3-81). Two patients recurred locally (one marginal and one in situ). Fifteen patients are alive and doing well. Overall, 12, 24, and 36-month local control rate was 92 +/- 8% and, 12, 24, and 36-month overall survival rates were 93 +/- 6%, 83 +/- 11%, and 83 +/- 11%, respectively. Clinical initial symptoms remained stable or subsided in all patients. Early toxicities were in the expected range. CONCLUSIONS: With a mean 27 months follow-up, protontherapy was well tolerated for doses upto 69 CGE and with an excellent local control rate. PMID- 12652621 TI - Squamous carcinoma of the larynx and hypopharynx in children: a distinct clinical entity? PMID- 12652620 TI - Results of p53 analysis in pediatric malignancies in Poland. AB - BACKGROUND: Mutations of the p53 gene are thought to be causally associated with the development of various neoplasms. In tumors overexpressing the wild-form of p53, its functional inactivation has been suggested, and MDM2 seems to be important in this process. We analyzed p53 in childhood solid tumors, as data on pediatric malignancies are still limited. PROCEDURE: The p53 gene was screened for mutations by the PCR-S SCP method and sequencing. p53, p21, and MDM2 proteins were analyzed by Western blotting. RESULTS: Overall, p53 mutations were found at a low frequency, 7% (9/126); the frequency calculated for sarcomas was also low, 8.6%. Interestingly, three of the nine detected mutations were new ones. p53 protein was demonstrated in all tumor histotypes, overall, in 63% (43/68) of the tumors, with 18% showing marked overexpression. No p21 was found; and the 76 kDa MDM2 protein was demonstrated in 18% (6/33) of the sarcomas. CONCLUSIONS: In the series of pediatric malignancies studied, the frequency of p53 mutations was very low, whereas p53 protein was present in a high fraction of the tumors. Thus, in total, p53 abnormalities were frequent. PMID- 12652623 TI - Adrenocortical carcinoma in children: a role for etoposide and cisplatin adjuvant therapy? Preliminary report. PMID- 12652624 TI - Hypothalamic dysfunction associated with neuroblastoma: evidence for a new Paraneoplastic syndrome? PMID- 12652625 TI - Management of critically ill children with mediastinal neoplasms: a 6-year survey from a single institution. PMID- 12652626 TI - Childhood haemangiopericytoma. PMID- 12652628 TI - Extraskeletal myxoid chondrosarcoma as a second malignancy after bone marrow transplantation. PMID- 12652627 TI - Successful immunosuppressive therapy for a child with Myelodysplastic syndrome. PMID- 12652629 TI - Relapse of childhood acute lymphoblastic leukemia in the lacrimal gland. PMID- 12652630 TI - Gastrointestinal stromal tumor in a patient surviving osteosarcoma. PMID- 12652631 TI - Highly conformal radiotherapy for craniopharyngioma: (potentially) throwing the baby out with the bathwater. PMID- 12652633 TI - Spirituality and the MMPI-2. AB - The present investigation was an exploratory examination of the relation of the Minnesota Multiphasic Personality Inventory-2 (MMPI-2; Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989) Clinical scales to spirituality operationalized in terms of self-reported religious involvement and scores on a multidimensional measure called the Expressions of Spirituality Inventory (ESI; MacDonald, 1997, 2000). MANOVA and correlational results indicate that the MMPI-2 Clinical scales generate patterns of findings consistent with available research on spirituality and health. In particular, persons reporting involvement in organized religion obtained significantly lower MMPI-2 Clinical scale scores and were found to be less likely to obtain a clinically significant score (i.e., t scores >64) on any of the MMPI-2 scales. Further, with the exception of Masculine Feminine and Hypomania, all MMPI-2 scales were found to associate appreciably with ESI dimension scores. The study concludes with a brief discussion of the meaning and implications of the findings for future research aimed at investigating the relation of spirituality to health. PMID- 12652634 TI - Perceived criticism predicts severity of anxiety symptoms after behavioral treatment in patients with obsessive-compulsive disorder and panic disorder with agoraphobia. AB - In prospective research, psychiatric patients' perceived criticism (PC) from family members has been linked to higher rates of relapse and worse treatment outcome. Researchers have disagreed about whether PC contributes to poor treatment outcomes or whether it merely reflects the severity of a patient's disturbance. In this study, structural equation modeling was used to conduct a cross-lagged panel analysis of the relationship between PC and anxiety symptom severity assessed before and after treatment in anxiety-disordered patients. PC was found to be unrelated to concurrent symptom severity. Furthermore, pretreatment PC significantly predicted posttreatment symptom severity over and above the effect of pretreatment symptom severity, whereas pretreatment symptom severity failed to significantly predict posttreatment PC. Thus, these results are consistent with (although not proof of) the hypothesis that PC detracts from patients' ability to respond to treatment, and inconsistent with the hypothesis that PC is a reflection of a patient's symptom severity. PMID- 12652635 TI - Use of the Coping Inventory for Stressful Situations in a clinically depressed sample: factor structure, personality correlates, and prediction of distress. AB - The Coping Inventory for Stressful Situations (CISS; Endler & Parker, 1990) is a self-report measure of Emotion-, Task-, and Avoidance-oriented coping. The psychometric properties of the CISS were evaluated in a large sample of outpatients with major depressive disorder (N = 298). The CISS scales demonstrated good reliability and support for their factorial validity was obtained. Relationships between the CISS scales and the broad personality domains from the five-factor model of personality, as well as two measures of emotional distress, were examined. Less-adaptive coping strategies (i.e., Emotion-oriented coping) were associated with less-adaptive personality traits (i.e., Neuroticism) and with psychological distress (i.e., Depression), whereas the reverse was found regarding adaptive coping strategies (i.e., Task-orientated coping). The incremental validity of the CISS was demonstrated by multiple-regression analyses that found two CISS scales accounted for significant variance in psychological distress beyond that contributed by the demographic and personality variables. PMID- 12652636 TI - Individualized measurement of irrational beliefs in remitted depressives. AB - Recent reviews of cognitive theories of depression have noted that individualized assessment strategies might help to resolve mixed findings regarding the stability of depressotypic beliefs and attitudes. We describe encouraging results for an individualized measure of one such cognitive construct, irrational beliefs. Twenty depression-prone women (recurrent major depressives in full remission) and twenty closely matched never-depressed controls completed leading forced-choice measures of irrational beliefs (the Belief Scale; BS) and sociotropy-autonomy (The Revised Personal Style Inventory), as well as the Specific Demands on Self Scale (SDS). The BS requires participants to rate their agreement with twenty preselected statements of irrational beliefs, while the SDS focuses on whether participants harbor any strongly held irrational beliefs, even if uncommon or idiosyncratic. Consistent with previous research, there were no group differences on the traditional measure of irrational beliefs. In contrast, depression-prone participants strongly exceeded controls on the SDS, and this difference persisted after controlling for residual depression, anxiety symptoms, anxiety diagnoses, sociotropy, and autonomy. These findings provide some initial support for a key assumption of the rational-emotive model of depression, and, more broadly, suggest that individualized assessment strategies may help researchers capture the core negative beliefs of asymptomatic individuals, even in the absence of mood or cognitive priming. PMID- 12652637 TI - Prediction of IQ in the Mayo Older Adult Normative sample using multiple methods. AB - Using the Mayo Older Adult Normative sample (Ivnik et al., 1992) as our database, we developed regression models for estimating premorbid Full Scale (FSIQs), Verbal (VIQs), and Performance (PIQs) IQs for elderly adults. Age, years of education, and sex were the only demographic variables that showed sufficient variability; therefore, they were used as predictor variables in three stepwise procedures. The Mayo Normative FSIQs, VIQs, and PIQs served as the dependent variables. Both education and sex added significantly to the accounting of variance of both FSIQ and VIQ ( p < .001), whereas education ( p < .001) and age ( p < .05) were significant predictors of PIQ. These models produced statistically significant multiple Rs of .54, .58, and .35 ( p < .0001), with standard errors of estimate of 9.02, 8.28, and 10.77 for FSIQ, VIQ, and PIQ, respectively. Estimated FSIQs generated with the present model and the model developed by Barona and colleagues (Barona, Reynolds, & Chastain, 1984) were compared. The correlation between estimated IQs was large, the mean difference between IQs was very small, the standard deviations were nearly equal, and the categorical distributions of the two were similar. Because the Barona model is likely to be familiar to most clinicians, these findings argue in favor of the continued use of the Barona model, even when assessing people older than the WAIS R normative sample. Extensions of these models to the WAIS-III also are discussed. PMID- 12652638 TI - The Depression-Arkansas scale: A validation study of a new brief depression scale in an HMO. AB - Recent trends in mental-health care have increased the need for practical depression instruments. The Depression-Arkansas (D-ARK), a brief, economical, multipurpose instrument, has been validated for assessing major depressive disorder (MDD) and depressive-symptom severity. Psychometric properties of the D ARK were compared with standard depression scales (Beck Depression Inventory and Geriatric Depression Scale) among 294 adult and 193 senior primary-care patients, respectively, and 163 patients enrolled in cognitive-behavioral depression classes. The severity scale displayed adequate internal reliability (coefficient alpha =.81-.86), high correlation with the BDI-2 (r =.78-.83) and GDS (r =.75), and similar factor structure to the BDI-2. The D-ARK was calibrated against the BDI-2 and GDS, providing familiar severity category cutpoints with the new instrument. This study yields further data supporting the reliability, validity, and practical utility of the D-ARK. PMID- 12652639 TI - Marlowe-Crowne Social Desirability Scale and short Form C: forensic norms. AB - Marlowe-Crowne Social Desirability Scale (MC) (Crowne & Marlowe, 1960) scores were collected on 1096 individuals involved in forensic evaluations. No prior publication of forensic norms was found for this instrument, which provides a measure of biased self-presentation (dissimulation). MC mean score was 19.42 for the sample. Also calculated was the score on Form C (MC-C) (Reynolds, 1982), and the mean for this 13-item scale was 7.61. The scores for the current sample generally are higher than those published for non-forensic groups, and statistical analysis indicated the difference was significant for both the MC and MC-C (d =.75 and.70, respectively, p <.001). Neither gender nor educational level proved to be significant factors in accounting for variance, and age did not appear to be correlated with scores. Group membership of subjects based on referral reason (family violence, abuse, neglect, competency, disability) was significant for both the MC and MC-C scores. Results suggest the MC or MC-C can be useful as part of a forensic-assessment battery to measure biased self presentation. PMID- 12652640 TI - A preliminary validation of the Positive and Negative Suicide Ideation (PANSI) inventory with normal adolescent samples. AB - The present study evaluated the factor structure, reliability, and validity of the Positive and Negative Suicide Ideation (PANSI; Osman, Gutierrez, Kopper, Barrios, & Chiros, 1998) inventory in a sample of high-school youths. The PANSI is designed as a measure of risk and protective factors related to suicidal behavior. Participants (114 boys and 103 girls) completed the PANSI and other self-report instruments. Results of the confirmatory factor analyses supported adequate fit of the 2-factor oblique model to the sample data. Both factor scales attained adequate levels of reliability. Boys and girls did not differ in their responses to the PANSI scales. The PANSI scale scores were associated with scores from related measures. Logistic-regression analyses were used to evaluate the contributions of the PANSI scale scores to differentiate between the study groups. Receiver Operating Characteristic (ROC) analyses, using data from the psychiatric suicide risk and high-school control youths, were used to identify cutoff scores of 1.63 and 3.33 for the PANSI-negative and PANSI-positive scales, respectively. PMID- 12652641 TI - Therapist use of silence in therapy: a survey. AB - Eighty-one therapists responded to a mailed survey about their use of silence during a specific event in therapy and about their general attitudes about using silence in therapy. For the specific event, therapists used silence primarily to facilitate reflection, encourage responsibility, facilitate expression of feelings, not interrupt session flow, and convey empathy. During silence, therapists observed the client, thought about the therapy, and conveyed interest. In general, therapists indicated that they would use silence with clients who were actively problem solving, but they would not use silence with very disturbed clients. Therapists learned about using silence mostly through clinical experience. PMID- 12652642 TI - Human brain synembryn interacts with Gsalpha and Gqalpha and is translocated to the plasma membrane in response to isoproterenol and carbachol. AB - Heterotrimeric G-proteins transduce signals from heptahelical transmembrane receptors to different effector systems, regulating diverse complex intracellular pathways and functions. In brain, facilitation of depolarization-induced neurotransmitter release for synaptic transmission is mediated by Gsalpha and Gqalpha. To identify effectors for Galpha-proteins, we performed a yeast two hybrid screening of a human brain cDNA library, using the human Galphas protein as a bait. We identified a protein member of the synembryn family as one of the interacting proteins. Extending the study to other Galpha subunits, we found that Gqalpha also interacts with synembryn, and these interactions were confirmed by in vitro pull down studies and by in vivo confocal laser microscopy analysis. Furthermore, synembryn was shown to translocate to the plasma membrane in response to carbachol and isoproterenol. This study supports recent findings in C. elegans where, through genetic studies, synembryn was shown to act together with Gqalpha regulating neuronal transmitter release. Based on these observations, we propose that synembryn is playing a similar role in human neuronal cells. PMID- 12652643 TI - Cell death regulation by the Bcl-2 protein family in the mitochondria. AB - An increase in the permeability of the outer mitochondrial membrane is central to apoptotic cell death, since it leads to the release of several apoptogenic factors, such as cytochrome c and Smac/Diablo, into the cytoplasm that activate downstream death programs. During apoptosis, the mitochondria also release AIF and endonuclease G, both of which are translocated to the nucleus and are implicated in apoptotic nuclear changes that occur in a caspase-independent manner. Mitochondrial membrane permeability is directly controlled by the major apoptosis regulator, i.e., the Bcl-2 family of proteins, mainly through regulation of the formation of apoptotic protein-conducting pores in the outer mitochondrial membrane, although the precise molecular mechanisms are still not completely understood. Here, I focus on the mechanisms by which Bcl-2 family members control the permeability of mitochondrial membrane during apoptosis. PMID- 12652644 TI - RET and NTRK1 proto-oncogenes in human diseases. AB - RET and NTRK1 are receptor tyrosine kinase (RTK) proteins which play a role in the development and maturation of specific component of the nervous system. Their alterations have been associated to several human diseases, including some forms of cancer and developmental abnormalities. These features have contributed to the concept that one gene can be responsible for more than one disease. Moreover, both genes encoding for the two RTKs show genetic alterations that belong to either "gain of function" or "loss of function" class of mutations. In fact, receptor rearrangements or point mutations convert RET and NTRK1 in dominantly acting transforming genes leading to thyroid tumors, whereas inactivating mutations, associated with Hirschsprung's disease (HSCR) and congenital insensitivity to pain with anhidrosis (CIPA), impair RET and NTRK1 functions, respectively. In this review we have summarized the main features of the two receptors, their physiological and pathological roles. In addition, we attempted to identify the correlations between the different genetic alterations and the related pathogenetic mechanisms. PMID- 12652645 TI - Voltage-dependent K+ channel beta subunits in muscle: differential regulation during postnatal development and myogenesis. AB - Voltage-dependent potassium channels contribute to the electrical properties of nerve and muscle by affecting action potential shape and duration. The complexity of the currents generated is further enhanced by the presence of accessory beta subunits. Here we report that while all Kvbeta mRNA isoforms are present in rat brain, muscle tissues express only Kvbeta1 (Kvbeta1.1-Kvbeta1.3) and Kvbeta2, but not Kvbeta3. Kvbeta subunits were close regulated through post-natal development in brain and striated muscle, as well as during myogenesis in the rat skeletal muscle cell line L6E9. While the alternatively spliced Kvbeta mRNA products from Kvbeta1 gene were differentially expressed, Kvbeta2.1 was associated with myogenesis. These results show that Kvbeta genes are strongly regulated in muscle and suggest a physiological role for voltage-gated K(+) channels during development and myotube formation. PMID- 12652646 TI - Mouse epidermal stem cells proceed through the cell cycle. AB - The epidermis is a continuously renewing tissue maintained by undifferentiated stem cells. For decades it has been assumed that epidermal stem cells (ESCs) were held in the G0 phase of the cell cycle and that they only entered the cell cycle when needed. Previously, we showed that ESCs retained nuclear label for long periods, indicating that these cells did not proceed through the cell cycle at the same rate as the other proliferative basal cells. However, their exact cell cycle profile has not been determined because a pure population of ESCs has not been available. In this study, we sorted stem and transient amplifying (TA) cells from murine neonatal back skin, and adult ear, footpad, and back skin, using our recently developed method. We found that neonatal back skin had two times the number of ESCs as the adult tissues. Despite the age and anatomical difference, these ESC populations exhibited similar cell cycle profiles with approximately 96% in G0/G1 and 4% in S-G2/M. The cell cycle profiles of the TA cells from neonatal back skin and adult footpad also showed a profile similar to each other (85% in G1 and 15% in S-G2/M). Examination of genes on a cell cycle chip showed that proliferation associated genes and only p57 were upregulated in the TA cell and ESC population, respectively. We found BrdU positive and cyclin B1 positive cells in all groups, confirming that both ESCs and TA cells were cycling. These data demonstrate that there are more TA cells dividing than ESCs, that the cell cycle profile of adult TA cells is related to the proliferative state of the tissue in which they reside, and that ESC proceed through the cell cycle. PMID- 12652647 TI - Involvement of S6 kinase and p38 mitogen activated protein kinase pathways in strain-induced alignment and proliferation of bovine aortic smooth muscle cells. AB - Bovine aortic smooth muscle cell (SMC) phenotype can be altered by physical forces. This has been demonstrated by cyclic strain-induced changes in proliferation and alignment. However, the intracellular coupling pathways remain ill defined. In the present study, we examined whether the p38 and S6 kinase pathway were involved in the mitogenic and morphological changes seen in SMCs exposed to cyclic strain. We seeded bovine aortic SMCs on silastic membranes that were deformed with 150-mmHg vacuum. Cyclic strain induced both alignment and proliferation of SMCs. SB202190, a specific inhibitor of p38, hindered SMC alignment, but not proliferation. Rapamycin, a specific inhibitor of the mTOR-S6 kinase pathway, attenuated strain-induced proliferation, but not alignment. Peak activation of p38 and S6 kinase was 351 +/- 76.9% at 5 min and 363 +/- 56.2% at 60 min compared with static control, respectively (P < 0.05). The results suggest that strain-induced SMC alignment is dependent on activation of p38, but not S6 kinase. Strain induced SMC proliferation is S6 kinase, but not p38 activation, dependent. PMID- 12652648 TI - Inhibition of protein kinase C decreases prostaglandin-induced breakdown of the blood-retinal barrier. AB - Breakdown of the blood-retinal barrier (BRB) occurs in several retinal diseases and is a major cause of visual loss. Vascular endothelial growth factor (VEGF) has been implicated as a cause of BRB breakdown in diabetic retinopathy and other ischemic retinopathies, and there is evidence to suggest that other vasopermeability factors may act indirectly through VEGF. In this study, we investigated the effect of several receptor kinase inhibitors on BRB breakdown resulting from VEGF, tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), insulin-like growth factor-1 (IGF-1), prostaglandin E1 (PGE(1)), or PGE(2). Inhibitors of VEGF receptor kinase, including PKC412, PTK787, and SU1498, decreased VEGF-induced breakdown of the BRB. None of the inhibitors blocked leakage caused by TNF-alpha, IL-1beta, or IGF-1 and only PKC412, an inhibitor of protein kinase C (PKC) as well as VEGF and platelet-derived growth factor (PDGF) receptor kinases, decreased leakage caused by prostaglandins. Since the other inhibitors of VEGF and/or PDGF receptor kinases that do not also inhibit PKC had no effect on prostaglandin-induced breakdown of the BRB, these data implicate PKC in retinal vascular leakage caused by prostaglandins. PKC412 may be useful for treatment of post-operative and inflammatory macular edema, in which prostaglandins play a role, as well as macular edema associated with ischemic retinopathies. PMID- 12652649 TI - EGF-responsive rat neural stem cells: molecular follow-up of neuron and astrocyte differentiation in vitro. AB - Neural stem cells (NSCs) could be very useful for the "cell therapy" treatment of neurological disorders. For this reason basic studies aiming to well characterize the biology of NSCs are of great interest. We carried out a molecular and immunocytochemical analysis of EGF-responsive NSCs obtained from rat pups. After the initial growth of NSCs as floating neurospheres in EGF-containing medium, cells were plated on poly-L-ornithine-coated dishes either in the presence or absence of EGF. We followed cell differentiation and apoptosis for 21 days in vitro and analyzed the expression levels of some genes having a major role in these processes, such as pRB, pRB2/p130, p27, and p53. We observed that EGF impairs neuronal differentiation. Furthermore, in the absence of mitogens, apoptosis, which appeared to proceed through the "p53 network," was significantly lower than in the presence of EGF. The cyclin kinase inhibitor p27, while important for cell cycle exit, seemed dispensable for cell survival and differentiation. PMID- 12652650 TI - Activation of P2Y2 receptor induces c-FOS protein through a pathway involving mitogen-activated protein kinases and phosphoinositide 3-kinases in HeLa cells. AB - The effects of P2Y2 purinoceptor activation on c-Fos expression and the signaling pathways evoked by extracellular ATP/UTP in HeLa cells were investigated. We found that P2Y2 activation induced c-Fos protein and phosphorylated the extracellular signal-regulated kinases 1 and 2 (ERK1/2). The P2Y2-stimulated c Fos induction was partly blocked (a) by U73122, a phospholipase C inhibitor, (b) by Go6976, a conventional PKC inhibitor, (c) by PD098059, a mitogen-activated protein kinase kinase inhibitor, and, moreover, (d) by the inhibitors of phosphoinositide 3-kinases (PI3K), LY294002 and wortmannin. When Go6976 and PD098059, or Go6976 and wortmannin, were combined there was a totally inhibition of P2Y2-induced c-Fos increase. Either U73122 or Go6976 did not inhibit ERK1/2 phosphorylation induced by ATP/UTP, while it was inhibited by LY294002 (or wortmannin) and by staurosporine. Additionally, wortmannin inhibited the cytosol to-membrane translocation of PKC- epsilon induced by ATP/UTP. These data indicated that agonist-induced PI3K and downstream PKC- epsilon activation mediated the effect of ATP/UTP on ERK1/2 activation. To test the biological consequences of ERK1/2 activation, the effect of P2Y2 on cell functions were examined. P2Y2 stimulation increased cell proliferation and this effect was attenuated by PD098059 in a dose-dependent manner, thereby indicating that the ERK pathway mediates mitogenic signaling by P2Y2. In conclusion, the activation of conventional PKCs through P2Y2 receptor acts in concert with ERK and PI3K/PKC- epsilon pathways to induce c-Fos protein and HeLa cell proliferation. PMID- 12652652 TI - Role of protein kinase C alpha in calcium induced keratinocyte differentiation: defective regulation in squamous cell carcinoma. AB - Calcium induces both involucrin and transglutaminase-K in normal keratinocytes (NHK) but not in squamous carcinoma cell lines (SCC). The protein kinase C (PKC) agonist phorbol myristoyl acetate potentiates and the PKC antagonist Ro31-8220 blocks the ability of calcium to stimulate the involucrin promoter in normal human keratinocytes but not in SCC4. We thus examined the ability of calcium to regulate the levels of five PKC isozymes in NHK and two SCC. In the normal keratinocytes, the levels of PKC [alpha], PKC [delta], PKC [eta], and PKC [zeta] increased over the first one to two weeks in a calcium-and time-dependent manner. PKC [epsilon] decreased in a time-and calcium-dependent fashion over the three week period. All five isozymes showed little change during culture in SCC4 at any calcium concentration. Calcium and time of culture had partial effects on SCC12B2, a carcinoma that shows partial differentiation characteristics. Since PKC [alpha] is the only calcium responsive PKC isozyme in keratinocytes and most likely to be directly involved in calcium induced differentiation, we evaluated the effect of inhibiting its production with antisense oligonucleotides on calcium-regulated markers of differentiation. We found that the PKC [alpha] specific antisense oligonucleotide blocked calcium stimulated involucrin promoter activity as well as PKC [alpha], involucrin, and transglutaminase protein production, whereas the sense oligonucleotide control did not. We conclude that although a number of PKC isozymes are regulated during calcium-induced differentiation, PKC [alpha] plays a necessary role in mediating calcium-induced differentiation. Failure to regulate PKC [alpha] in SCC4 may underlie at least part of the failure of calcium to promote differentiation in these cells. PMID- 12652651 TI - VEGF-TRAP(R1R2) suppresses choroidal neovascularization and VEGF-induced breakdown of the blood-retinal barrier. AB - Vascular endothelial growth factor (VEGF) plays a central role in the development of retinal neovascularization and diabetic macular edema. There is also evidence suggesting that VEGF is an important stimulator for choroidal neovascularization. In this study, we investigated the effect of a specific inhibitor of VEGF, VEGF TRAP(R1R2), in models for these disease processes. VEGF-TRAP(R1R2) is a fusion protein, which combines ligand binding elements taken from the extracellular domains of VEGF receptors 1 and 2 fused to the Fc portion of IgG1. Subcutaneous injections or a single intravitreous injection of VEGF-TRAP(R1R2) strongly suppressed choroidal neovascularization in mice with laser-induced rupture of Bruch's membrane. Subcutaneous injection of VEGF-TRAP(R1R2) also significantly inhibited subretinal neovascularization in transgenic mice that express VEGF in photoreceptors. In two models of VEGF-induced breakdown of the blood-retinal barrier (BRB), one in which recombinant VEGF is injected into the vitreous cavity and one in which VEGF expression is induced in the retina in transgenic mice, VEGF-TRAP(R1R2) significantly reduced breakdown of the BRB. These data confirm that VEGF is a critical stimulus for the development of choroidal neovascularization and indicate that VEGF-TRAP(R1R2) may provide a new agent for consideration for treatment of patients with choroidal neovascularization and diabetic macular edema. PMID- 12652653 TI - Heregulin and retinoids synergistically induce branching morphogenesis of breast cancer cells cultivated in 3D collagen gels. AB - C-erbB and retinoid receptor signaling control mammary epithelial cell proliferation, differentiation, and morphology. Here, we examined the morphogenetic activities of c-erbB specific ligands such as heregulin and of retinoids on non-malignant (primary, MTSV1-7) and malignant (T47D, SKBR-3) human mammary epithelial cells (HMEC) cultivated in 3D collagen type I gels. These cells are positive for both c-erbB and retinoid receptors. Non-malignant primary HMEC spontaneously formed branched structures in collagen, whereas SV40 large T antigen-immortalized non-tumorigenic MTSV1-7 spontaneously formed balls and required heregulin or retinoid X receptor alpha-selective retinoid Ro 25-7386 for branching, which was further stimulated by combination of both types of agents. In malignant cells, heregulin alone induced ball formation and cooperated either with Ro 25-7386 (T47D) or with retinoic acid receptor alpha-selective AM580 (SKBR 3) for branching morphogenesis, which was accompanied by changes in the subcellular distribution of alpha(2)beta(1)-integrin and E-cadherin, and by down regulation of c-erbB-2, -3, or -4. Heregulin and/or retinoids correspondingly increased the integrin-dependent adhesion of malignant cells to type I collagen. Our data demonstrate cooperative signaling of c-erbB and retinoid receptor pathways at the levels of morphogenesis and immunophenotypic differentiation. PMID- 12652655 TI - Activation of extracellular signal-regulated kinases ERK1 and ERK2 induces Bcl-xL up-regulation via inhibition of caspase activities in erythropoietin signaling. AB - Erythropoietin (EPO) can rescue erythroid cells from apoptosis during erythroid development, leading to red cell production. However, the detailed mechanism of how EPO protects erythroid cells from apoptosis is still open to question. To address this problem, we used a human EPO-dependent leukemia cell line UT-7/EPO and normal erythroid progenitor cells. After deprivation of EPO, UT-7/EPO cells underwent apoptosis, accompanied by down-regulation of the Bcl-xL protein. In addition, the cleaved products of caspase-3, p11 and p21, and a few cleaved forms of inhibitor of caspase-activated DNase (ICAD) were detected in these cells. When the cells were pre-treated with the pancaspase inhibitor Z-VAD-FMK, the ratio of apoptotic cells was significantly reduced, suggesting that EPO protects the UT 7/EPO cells from apoptosis via inhibition of caspase activities. When an MEK 1/2 inhibitor U0126 inhibited activities of extracellular signal-regulated kinases (ERKs), the expression of Bcl-xL protein was down-regulated and subsequently apoptosis was induced. Interestingly, Z-VAD-FMK blocked U0126-induced down regulation of Bcl-xL protein and apoptosis, strongly suggesting that Bcl-xL expression is regulated by caspases which lies downstream of ERK activation pathway in EPO signaling. Importantly, these findings were also observed in normal erythroid progenitor cells. In conclusion, the activation of ERKs by EPO up-regulates Bcl-xL expression via inhibition of caspase activities, resulting in the protection of erythroid cells from apoptosis. PMID- 12652654 TI - Expression and tyrosine phosphorylation of Cbl regulates macrophage chemokinetic and chemotactic movement. AB - Primary macrophages isolated from hck(-/-)fgr(-/-) mice display altered morphology and F-actin cytoskeletal structures and reduced migration. The ability of phorbol myristyl acetate (PMA), a protein kinase C activator that has been reported to increase macrophage spreading and carcinoma cell motility, to rescue these hck(-/-)fgr(-/-) defects was tested. Although PMA-treated wild-type and hck(-/-)fgr(-/-) macrophages exhibited a similar flattened, spread phenotype, PMA did not rescue the hck(-/-)fgr(-/-) macrophage migration defect. Instead, both PMA-treated wild type and hck(-/-)fgr(-/-) macrophages were defective in spontaneous and chemotactic migration and tyrosine phosphorylation of the Cbl protooncoprotein was decreased in both. Moreover, c-cbl(-/-) macrophages displayed the same impairment of motility as hck(-/-)fgr(-/-) macrophages and a similar morphology with less polarization and more dorsal ruffling than wild-type macrophages. As Hck and Fgr expression and activity were not decreased in c-cbl( /-) macrophages, these results suggest that Cbl is likely to be an important downstream mediator of the Src family kinase-regulated macrophage motility pathway. PMID- 12652656 TI - Differential effects of quercetin, a natural polyphenolic flavonoid, on L-type calcium current in pituitary tumor (GH3) cells and neuronal NG108-15 cells. AB - The effects of quercetin, a natural polyphenolic compound, on voltage-dependent L type Ca(2+) current (I(Ca,L)) in rat pituitary GH(3) cells were investigated with the aid of the whole-cell voltage-camp technique. Quercetin (0.5-200 microM) stimulated I(Ca,L) in a concentration-dependent manner. The current-voltage (I-V) relationship of I(Ca,L) was slightly shifted to more negative potentials in the presence of quercetin. The EC(50) value of the quercetin-induced stimulation of I(Ca,L) was about 7 microM. The presence of quercetin (5 microM) shifted the steady state inactivation curve of I(Ca,L) to a more negative potential by approximately -10 mV. Although quercetin might increase intracellular cyclic AMP, sp-cAMPS did not affect I(Ca,L). In addition, neither flavone nor wortmannin had any effect on the amplitude of I(Ca,L), while epicatechin and genistein slightly suppressed it. Quercetin (50 microM) decreased the amplitude of tetrodotoxin sensitive Na(+) current in GH(3) cells. Under current-clamp configuration, quercetin could increase the firing frequency of actions potentials. Conversely, in NG108-15 neuronal cells, quercetin suppressed the amplitude of I(Ca,L). The quercetin-induced inhibition of I(Ca,L) was abolished in NG108-15 cells preincubated with t-butyl hydroperoxide (1 mM). Quercetin-mediated stimulation of I(Ca,L) in GH(3) cells was presumably not associated with the level of intracellular cyclic AMP, or with the activity of tyrosine or phosphoinositide 3 kinases. Therefore, the effects of quercetin on ion currents may, at least in part, contribute to the underlying mechanisms through which it affects neuronal or neuroendocrine function. PMID- 12652657 TI - Identification of cathepsin K as a novel marker of adiposity in white adipose tissue. AB - In obesity, adipocytes undergo dramatic morphological and molecular changes associated with alterations in their gene expression profile. To identify genes differentially modulated in white adipose tissue (WAT) of obese db/db mice compared to wild type (wt) mice, we utilized RNA fingerprinting. Among the 52 candidates that we identified, we focused here on cathepsin K (ctsk), a cysteine protease, prevalently localized in lysosomes and involved in bone extracellular matrix degradation. In db/db mice, WAT ctsk mRNA was elevated 5.9-fold, as were Mitf and TFE3 (2- and 3.3-fold respectively), two transcription factors involved in ctsk induction in osteoclasts. Moreover, the level of WAT ctsk mRNA was increased in other obese models including A(y), fat, and tubby (2.8-, 3.2-, and 4.9-fold respectively) and decreased in mice undergoing weight loss. Despite the ubiquitous distribution of the ctsk transcript, we demonstrated that the obesity related increase is specific to the adipocytes. Further, in vitro experiments proved that the abundance of ctsk transcript increases upon adipose conversion of the established cell line of preadipocytes 3T3-F442A. In addition, ctsk gene expression was examined in adipose tissue of 21 lean and obese male subjects and significant correlations with BMI (r = 0.54, P = 0.012) and plasma leptin levels (r = 0.54, P = 0.015) were found. In conclusion, the WAT of obese db/db mice exhibits a different expression profile from that of the wt mice, and cathepsin K can be considered a novel marker of obesity and a target for the inhibition of adipose mass growth. PMID- 12652659 TI - A matter of survival? PMID- 12652658 TI - Beta-adrenergic receptor-mediated DNA synthesis in neonatal rat cardiac fibroblasts proceeds via a phosphatidylinositol 3-kinase dependent pathway refractory to the antiproliferative action of cyclic AMP. AB - The following study was undertaken to elucidate the cytoskeletal phenotype of neonatal rat cardiac fibroblasts (NNCF) and the signaling pathways coupled to beta-adrenergic receptor stimulated DNA synthesis. The cytoskeletal proteins vimentin, and smooth muscle alpha-actin were detected in NNCF, suggestive of a myofibroblast phenotype. Isoproterenol (ISO) treatment stimulated (3)H-thymidine uptake, and concomitantly increased intracellular cyclic AMP levels. However, cyclic AMP-elevating agents markedly decreased DNA synthesis. Coincident with growth, ISO-stimulated phosphatidylinositol 3-kinase (PI3-K) activity, and the PI3-K inhibitor LY294002 abrogated enzyme activity, and DNA synthesis. Unexpectedly, the serine/threonine kinase protein kinase Balpha (PKBalpha), a putative downstream target of PI3-K, was dephosphorylated following ISO treatment. Despite PKBalpha inactivation, the phosphorylation of its putative downstream target, the pro-apoptotic enzyme glycogen synthase kinase-3alpha was significantly increased in response to ISO. These latter effects of ISO were mimicked by the cyclic AMP-elevating agent forskolin. Lastly, ISO treatment increased p70 ribosomal S6 kinase (p70S6K) phosphorylation, as reflected by an upward electrophoretic mobility shift. The pretreatment with rapamycin abrogated the ISO-mediated mobility shift of p70S6K, and DNA synthesis. Collectively, these data demonstrate that NNCF express a myofibroblast phenotype, and beta-adrenergic agonists promote DNA synthesis via a PI3-K-dependent pathway involving p70S6K. Although unable to suppress ISO-stimulated DNA synthesis, cyclic AMP can influence specific downstream targets of PI3-K highlighting a novel crosstalk between these signaling pathways. PMID- 12652660 TI - Synergistic science. AB - Should computational materials science be recognized as a field with a role in the community comparable to computational physics or chemistry? With the emergence of multiscale modelling, the answer is a resounding 'yes'. PMID- 12652661 TI - Controlling plastic instability. PMID- 12652662 TI - Self-organization of functional polymers. PMID- 12652663 TI - Harnessing the unstable. PMID- 12652664 TI - Switching light with light. PMID- 12652665 TI - Tougher ceramics with nanotubes. PMID- 12652666 TI - Framework under pressure. PMID- 12652667 TI - Conducting polymers in electronic chemical sensors. AB - Conducting organic polymers have found two main kinds of application in electronics so far: as materials for construction of various devices and as selective layers in chemical sensors. In either case, interaction with ambient gases is critical. It may compromise the performance of a device based on conducting polymers, whereas it is beneficial in a sensor. Conductivity has been the primary property of interest. Work function--related to conductivity, but in principle a different property--has received only scant attention. Our aim here is to discuss the usability of conducting polymers in both types of electronic applications in light of these two parameters. PMID- 12652668 TI - Stainless steel optimization from quantum mechanical calculations. PMID- 12652669 TI - Multicolour photochromism of TiO2 films loaded with silver nanoparticles. PMID- 12652670 TI - Novel Ti-base nanostructure-dendrite composite with enhanced plasticity. AB - Single-phase nanocrystalline materials undergo inhomogeneous plastic deformation under loading at room temperature, which results in a very limited plastic strain (smaller than 0-3%). The materials therefore display low ductility, leading to catastrophic failure, which severely restricts their application. Here, we present a new in situ-formed nanostructured matrix/ductile dendritic phase composite microstructure for Ti-base alloys, which exhibits up to 14.5% compressive plastic strain at room temperature. The new composite microstructure was synthesized on the basis of the appropriate choice of composition, and by using well-controlled solidification conditions. Deformation occurs partially through dislocation movement in dendrites, and partially through a shear-banding mechanism in the nanostructured matrix. The dendrites act as obstacles restricting the excessive deformation by isolating the highly localized shear bands in small, discrete interdendritic regions, and contribute to the plasticity. We suggest that microscale ductile crystalline phases might therefore be used to toughen nanostructured materials. PMID- 12652671 TI - Single-wall carbon nanotubes as attractive toughening agents in alumina-based nanocomposites. AB - The extraordinary mechanical, thermal and electrical properties of carbon nanotubes have prompted intense research into a wide range of applications in structural materials, electronics, chemical processing and energy management. Attempts have been made to develop advanced engineering materials with improved or novel properties through the incorporation of carbon nanotubes in selected matrices (polymers, metals and ceramics). But the use of carbon nanotubes to reinforce ceramic composites has not been very successful; for example, in alumina-based systems only a 24% increase in toughness has been obtained so far. Here we demonstrate their potential use in reinforcing nanocrystalline ceramics. We have fabricated fully dense nanocomposites of single-wall carbon nanotubes with nanocrystalline alumina (Al2O3) matrix at sintering temperatures as low as 1,150 degrees C by spark-plasma sintering. A fracture toughness of 9.7 MPa m 1/2, nearly three times that of pure nanocrystalline alumina, can be achieved. PMID- 12652672 TI - Dynamics of ferroelastic domains in ferroelectric thin films. AB - Dynamics of domain interfaces in a broad range of functional thin-film materials is an area of great current interest. In ferroelectric thin films, a significantly enhanced piezoelectric response should be observed if non-180 degrees domain walls were to switch under electric field excitation. However, in continuous thin films they are clamped by the substrate, and therefore their contribution to the piezoelectric response is limited. In this paper we show that when the ferroelectric layer is patterned into discrete islands using a focused ion beam, the clamping effect is significantly reduced, thereby facilitating the movement of ferroelastic walls. Piezo-response scanning force microscopy images of such islands in PbZr0.2Ti0.8O3 thin films clearly point out that the 90 degrees domain walls can move. Capacitors 1 microm2 show a doubling of the remanent polarization at voltages higher than approximately 15 V, associated with 90 degrees domain switching, coupled with a d33 piezoelectric coefficient of approximately 250 pm V-1 at remanence, which is approximately three times the predicted value of 87 pm V-1 for a single domain single crystal. PMID- 12652673 TI - Hierarchical structure formation and pattern replication induced by an electric field. AB - Several techniques based on soft lithography have emerged to replicate micrometre sized patterns. Similar to most other lithographic methods, these techniques structure a single layer of photo resist. For many applications, however, it is desirable to control the spatial arrangement of more than one component. With traditional methods, this requires an iterative, multistep procedure, making the replication process more complex and less reliable. Here, a replication process is described where multiple materials are processed simultaneously. Using a bilayer formed by two different polymers, electrohydrodynamic instabilities at both polymer surfaces produce a hierarchic lateral structure that exhibits two independent characteristic dimensions. A lateral modulation of the electric field enables replication with a resolution down to 100 nanometres. This approach might provide a simple strategy for large-area, sub-100-nanometre lithography. PMID- 12652674 TI - A highly crystalline layered silicate with three-dimensionally microporous layers. AB - Layered silicates with three-dimensional microporosity within the layers have the potential to enable new applications in catalysis, adsorption and ion-exchange. Until now no such materials have been reported. However, here we present the synthesis and structure of AMH-3, a silicate with three-dimensionally microporous layers, obtained in high purity and crystallinity. AMH-3 is composed of silicate layers containing eight-membered rings in all three principal crystal directions, and spaced by strontium cations, sodium cations and water molecules. Because of its three-dimensional pore structure, acid and thermal stability, this layered material could find applications in polymer-silicate composites for membrane applications, for synthesis of combined microporous-mesoporous materials, and for the formation of new zeolites and microporous films. Its existence also opens new possibilities for the synthesis of other layered silicates with multidimensional microporous framework layers. PMID- 12652675 TI - Complex dewetting scenarios captured by thin-film models. AB - In the course of miniaturization of electronic and microfluidic devices, reliable predictions of the stability of ultrathin films have a strategic role for design purposes. Consequently, efficient computational techniques that allow for a direct comparison with experiment become increasingly important. Here we demonstrate, for the first time, that the full complex spatial and temporal evolution of the rupture of ultrathin films can be modelled in quantitative agreement with experiment. We accomplish this by combining highly controlled experiments on different film-rupture patterns with computer simulations using novel numerical schemes for thin-film equations. For the quantitative comparison of the pattern evolution in both experiment and simulation we introduce a novel pattern analysis method based on Minkowski measures. Our results are fundamental for the development of efficient tools capable of describing essential aspects of thin-film flow in technical systems. PMID- 12652676 TI - [Genetic education in pre- and post-graduation]. AB - Molecular genetics is now integral to all aspects of biomedical science. Every medical school should have a best program of basic and practical genetics education as one of the core curriculum. Genetic information includes DNA, chromosome analysis and clinical tests, as well as family history. The benefit of patient received genetic tests is counseling by counselors well educated. Thus each medical and technological school must find the best way to incorporate genetics teaching into its own curriculum. PMID- 12652677 TI - [Present situation of the genetic medical treatment]. AB - Medical treatment has been changing due to the recent and great progress in the molecule genetics. Gene tests are contained in the ordinary medical treatments. However, this progress in Japan has apparently thrown the general population into confusion, at least in part because of the lack of consent between inhabitants and clinicians/researchers. Some of the genetic diseases are difficult to be cured completely, although they can be diagnosed. Furthermore, genetic diseases spread the influences to other relations including their offspring. The preparation of the various genetic guidelines and the fulfillment of the genetic counseling are going to proceed to keep the human rights of the patients of the genetic diseases. A discussion of the bio-ethical problems has been developing at the same time. The necessity against the genetic service is increasing gradually. Genetic medical treatment may be a part of the genetic service. I report here the current status of "the genetic counseling and the genetic medical treatment" have been advanced in Nagasaki university hospital as an example. It is very important and expected that a proper genetic service is provided to the clients under the network of the specialists of the various fields. PMID- 12652678 TI - [External quality assessment of the genetic testings for hematopoietic tumor, CML]. AB - Genetic testings are commonly employed in various fields of clinical medicine and the test items performed at clinical laboratories are increasing rapidly in number. They are utilized to make early and/or definite diagnoses of infectious diseases, leukemia, cancers and molecular inherited diseases and also to monitor the progress of the diseases. However, these genetic testings except for infectious diseases have been developed independently at each clinical laboratory and the test results obtained at each laboratory are not always compatible each other. Under these situations it is widely expected to construct advanced genetic testing systems that can supply standardized data at any of domestic and international clinical laboratories. For the period from April, 1999 to March, 2002 three major clinical laboratories, SRL, Inc., BML, Inc. and MBC, Inc., were consigned by JBA (Japan Bioindustry Association) to collaborate in standardizing the evaluation methods for genetic testing systems among the clinical laboratories. The aim of the study is to develop the standardized genetic testing systems and to propose them as international standard operational procedures to the ISO/TC212 working group. Although one of the most important issues for standardization is the external quality assessment, they have not been carried out in reality. In this study we evaluated the difference of the genetic testing results obtained during the year of 2000 and 2001 among the clinical laboratories. The genetic testings for hematopoietic tumor, CML were selected to be evaluated since they are widely accepted as clinically useful tests. PMID- 12652679 TI - [Present and future prospects of individualized medical services by genetic tests]. AB - With progress of human genome projects many gene mutations and their expression profiles have been much identified. Laboratory tests have become essential to achieve proper individualized medical services. In particular single nucleotide polymorphisms (SNPs) and gene expression profiles are of high interests. As one of these ongoing mmodels, we here present the SNPs of human Protein 1' also called as human Clara cell 10-kd protein (CC10). We initially found six different SNPs (A-908G, G38A, G118A, C1225T, G1226A, C4777G). For laboratory use multiplex PCR was developed to determine frequencies in normal control. The -908G and 38A genotypes were more frequent in sarcoidosis than those in controls. The SNPs of in the human protein 1 gene may be the genetic susceptibility factors for Sarcoidosis. PMID- 12652680 TI - [Current status of gene test market]. AB - The technological innovation of the gene analysis makes the adaptation range of the gene test in clinical diagnosis expand. Then, gene test has popularized increasingly around the infection disease for clinical inspection. Also in the field of clinical inspection, the increase of the importance of clinical application and the inspection item new year by year have appeared with the functional analysis of a gene. Moreover, the new test method and automation analysis equipment tend to be developed by progress of gene-analysis technology, and it is going to be introduced. The spread of gene test and development of a gene test market have an important possibility of activating the present clinical inspection field. PMID- 12652681 TI - [Molecular diagnostic tests availability in view of remuneration for medical services]. AB - By the remarkable development of recent molecular biology and genetic technology, the molecular diagnostic tests had put into practice as the routine test in the field of the clinical laboratory examinations. For molecular diagnostic tests, there are mainly infectious disease, hematological malignancies, hereditary disease, cancers, etc. In especially infectious disease and hematological malignancies, the adaptation of health insurance are examined in the facilities. It is usefully utilized for early detection, early diagnosis, selection of the treatment, follow-up, especially in the hematological malignancies such as leukemia and malignant lymphoma. It is possible to determine the clonal origin of malignant cells, and to detect minimal residual disease (MRD) after achieving complete remission. The usage of the molecular diagnostic tests is variable in infectious disease and hematological malignancies on the medical reimbursement, and there are restrictions like the following; in the case of any tests of the infectious disease, only which or main test can be calculated, facilities standards are necessary in order to calculate the health insurance treatment cost of the once in 6 months. In this article, we describe utilization and restrictions of the medical reimbursement for molecular diagnostic tests, focusing on the fiscal 2002 revision of the medical service fee schedule implemented in April. PMID- 12652682 TI - [Biochemical marker for myocardial injury]. AB - Recently, the general idea of the acute coronary syndrome (ACS) that includes acute myocardial infarction and unstable angina has been established. ACC/AHA guidelines in 1996 and 1999 had emphasized the importance of the initial diagnosis and the early treatment in acute myocardial infarction. We presented about the recent advances in "injured myocardial marker" around the ACS in this paper. The sensitivity and specificity for detecting myocardial injury by troponin T and I(TnT and TnI) were superior compared with those in creatinine kinase (CK) and CK-MB. Easy rapid detection system by using whole blood was useful in the diagnosis a high risk cases in ACS. Measurement of TnT and TnI are recommended the use in the emergency room. A guideline for a diagnosis of acute myocardial infarction had a revision (ACC/ESC) on 2000. Biochemical markers of TnT & TnI were recommended as the first step for defining the myocardial injury. Unstable angina pectoris with a minimal myocardial injury (rise in CK under two times) was defined acute myocardial infarction. As for the measurement of the myocardial injury, rapid detection and accuracy & specificity may enhance the value of modality. In future, inflammation marker such as the high sensitivity CRP and an activated platelet may be thought to become important. PMID- 12652683 TI - [Is respiratory syncytial virus one of the causative agents for transient hyperphosphatasemia?]. AB - OBJECTIVE: In the winter of 2001, we found several cases of transient hyperphosphatasemia (TH) in patients with respiratory syncytial virus (RSV) infection. Therefore, we tried to verify that RSV was one of the causative agents for TH. RESULTS: From November 2001 to January 2002, we diagnosed 94 cases of RSV infection with the detection of virus antigens in the nasal mucus. Among these cases, we found six cases (6.4%) of TH, and all six patients were over one year of age. There were 55 cases of RSV infection in patients over one year of age, and the rate of TH with RSV infection was estimated to be 10.9% in this age group. This rate of TH (6.4%) was significantly higher than rates reported in previous studies (0.33% to 1.5%, p = 0.0145 to p < 0.0001). CONCLUSION: We conclude that RSV is one of the causative agents for TH. PMID- 12652685 TI - [Reference value and reference interval in laboratory medicine--official proposal of the Japanese Society of Laboratory Medicine]. PMID- 12652684 TI - [Elimination of electrocardiogram artifacts from electroencephalogram records by using the simultaneously recorded electrocardiograph data]. AB - We developed a simple method to eliminate electrocardiogram (ECG) artifacts from electroencephalogram (EEG) records by using simultaneously recorded ECG data. The raw EEG data, the real EEG data and the ECG data were regarded as multi dimensional vectors Ea, Er and C, respectively. Also, the ECG data, with reduced amplitude whose coefficient was denoted as 'k', were assumed to be overlapped on the real EEG. These assumptions introduced the equations [Ea = Er + k.C], [Er.C = 0] and finally [k = Ea. C/C.C]. This calculation method was implemented by a Macintosh computer using data exported from digital EEG recordings (sampled at 200 Hz with 16-bit resolution). In several subjects, sampling intervals of 5 or 10 seconds for calculation succeeded in eliminating ECG artifacts. However, regardless of the sampling interval, this elimination condition was not always efficient in several other subjects, including a brain-dead patient. It was suggested that the ECG data used were insufficient for the calculation, because only one hand-to-hand reference was used for simultaneous recording, as usual. This one ECG reference was able to express only one ECG projection. Then two other hand-to-foot references of ECG were added to the recordings, and the elimination procedure was performed using all of the simultaneously recorded ECG data at the three references. Consequently, elimination was much improved in most subjects, including the brain-dead patient. Our method may be useful for eliminating ECG artifacts without changing reference electrodes. PMID- 12652686 TI - [Clinical significance of MMP-3 in patients with rheumatoid arthritis: comparison with other inflammatory markers(IL-6, IL-8)]. AB - We determined serum metalloproteinase-3(MMP-3) and inflammatory cytokine(IL-6, IL 8) levels in patients with rheumatoid arthritis(RA). Sera were obtained from 307 healthy subjects(female 140, male 167), 54 RA patients, and 17 osteoarthritis (OA). The MMP-3 concentrations in healthy female and male were 43.3 +/- 15.3 ng/ml and 90.7 +/- 26.0 ng/ml, respectively. The serum MMP-3 levels in male were significantly higher than those in female (p < 0.0001). MMP-3 levels in RA patients(259.1 +/- 34.2 ng/ml) were significantly higher than OA(43.6 +/- 6.1 ng/ml) or healthy controls. There was a significant correlation between MMP-3 and CRP(r = 0.586), IL-6(r = 0.345) levels in serum. In contrast, no significant correlation was observed between MMP-3 and IL-8(r = 0.19), or CA-RF(r = 0.052) levels. However, there were some cases with high MMP-3 levels in CA-RF-negative patients definitely diagnosed as RA. These findings suggest that MMP-3 determination is useful for the early diagnosis and the follow-up during the treatment for RA patients. PMID- 12652687 TI - [Critical appraisal of evidence on diagnostic tests]. AB - The article explains the use of stratum-specific likelihood ratios(SSLRs) and the use of a checklist for critically appraising articles on diagnostic tests. The author recommends the use of SSLRs over sensitivity/specificity when evaluating the test characteristics of a given test, because the former retains as much information as possible of the test. Critical appraisal of a diagnostic test forms a part of the practice of evidence-based medicine(EBM). The particular importance of paying attention to work-up bias and spectrum bias when evaluating a diagnostic test is emphasized. It is hoped that future studies in laboratory medicine will follow these guidelines. PMID- 12652688 TI - [Why does EBD need to diagnose and to therapy for patients, now?]. AB - G. Guyatt reported the first paper on Evidence Based Medicine(EBM) at 1991. Everybody could understand rapidly the necessary of EBM to diagnose and medical treat the patients from then. So, I will tell you the recent actions of EBLM committee of Japan Society of Clinical Laboratory Medicine (JSLM/C-EBLM) related with EBM. I define EBLM(Evidence-Based Laboratory Medicine) has the total functions to use the clinical laboratory data by evidence for the diagnosis and the therapy of patients. JSLM/C-EBLM was seted up at 1999 and supported EBD(Evidence-Based Diagnosis) Forums as the main action. JSLM/C-EBLM will make and use practically the new meta-analysis tools and educate about EBLM from now. PMID- 12652689 TI - [Detection and identification of variant and abnormally modified proteins by soft ionization mass spectrometry, unrivaled technique for clinical diagnosis]. AB - Using the unique preparation and detection techniques we devised for application of proteins to soft ionization mass spectrometry, we found many variant and abnormally modified proteins. Up to end of 2002, we have diagnosed 96 cases of variant hemoglobins(Hb), which include 45 different and 3 new species. We have diagnosed or confirmed the molecular weight difference of 64 cases of variant transthyretins (TTR), which include 14 different and 3 new species. We found the first 2 cases showing homozygote of TTR 119Thr-->Met, a high frequent non-amyloid variant among some European ethnicities. Variants of Cu, Zn-superoxide dismutase, 5 cases, 5 different species, were analyzed and the differences of molecular weight suggested by DNA analyses were confirmed. These are listed in the text. Cysteine 10 in TTR is modified in vivo and in vitro. We found a marked increase in s-sulfonated TTR in 4 patients with molybdenum cofactor deficiency and homocysteine conjugated TTR in 2 patients with methylene H4 folate reductase deficiency. The carbohydrate deficient transferrin was detected in 3 cases of carbohydrate deficient glycoprotein syndrome. The detection of these modified structures might be expected to be a new diagnostic index for these diseases, which are easily misdiagnosed. The improved reference method for HbA1C measurement was established using isotope(deuterium) labeled glycated and non glycated hexapeptides from amino terminal of Hb beta-chain. We checked the discrepancies of HbA1c values by the routine method and the reference method we established. Most samples containing variant Hbs did not show correct HbA1c values by HPLC and some did not show correct values by immunoassay. We showed a typical example of an enormous increase in the glycation degree because of the amino acid substitution, which was shown clearly by electrospray ionization/mass spectrometry. (Nine figures and 4 tables were selected from 45 slides presented in the lecture.) PMID- 12652690 TI - [Present state of transfusion errors]. AB - Most serious transfusion error is ABO-incompatible transfusion. The Japanese Society of Blood Transfusion reported the results of the National Survey on the present state of ABO-incompatible blood transfusion in Japan. The targets of the survey were 777 hospitals with more than 300 beds, which have transfused more than 3,000 units of blood products per year. The answer could be obtained from 578(74%) hospitals among them. The questionnaire focused on the presence or absence of ABO-mismatched transfusion between January 1995 and December 1999. 115 hospitals(20%) have experienced ABO-mismatched transfusion in the period of the study. The frequency of ABO-mismatched transfusion increased with increasing the number of beds and the units of transfused blood. The ABO-mismatched transfusions were due to clerical or technical errors. The main causes of errors were misidentification of blood bags(42.8%), incorrect blood typing(15.1%) and failure to identify patient (11.5%). Incorrect blood typing errors were performed in 25 cases, and were caused by doctors in 17 cases, others by laboratory technicians. Majority of incorrect blood typing occurred during holidays or night shifts, and in emergency. PMID- 12652691 TI - [Necessity of a 24-hour system of blood transfusion testing]. AB - The preventive effects of a 24-hour system of blood transfusion testing on mistyping of transfused blood was examined. Blood transfusion tests have been performed by blood transfusion technologists during working hours and by physicians at other times. In March 2000, we introduced a system in which technologists perform blood transfusion tests after working hours. Technologists of the Blood Transfusion Unit and Central Clinical Laboratory perform the test jointly, and column agglutination technology was introduced as the test method. A computer system setup exclusively for the testing was also introduced to perform computer cross-matching. Since transfusion error is likely to occur during emergency blood transfusion, a manual was established to prioritize safety. After introduction of the system, mistyping that may have been caused by inaccurate blood test results markedly decreased, confirming the usefulness of this system for prevention of mistyping. In addition, transfusion errors also decreased in wards and the improved system increased the safety of the entire medical care system. The frequency of mistyping was about 1% when physicians performed blood typing, showing the importance of clinical technologists for blood transfusion tests. PMID- 12652692 TI - [24-hour system of the blood transfusion services at the university hospital]. AB - Blood transfusion is an essential medical treatment, from the everyday use in surgery to the treatment of large-scale bleeding accompanying external injury. Securing sufficient quantities of blood product and ensuring safe storage are critical. A 24-hour system of blood transfusion inspection by medical technologists has been enforced in our hospital since April, 1999 to improve the safety of blood transfusion treatment at night and on weekends. The medical technologists in the clinical laboratory and the division of blood transfusion carried out examinations in combination. There were no clinical problems after blood transfusion, and a safe blood supply has been maintained. As for medical technologists not specializing in the division of blood transfusion, blood type and crossmatching tests could be confirmed by the introduction of the Micro Typing System without problem. The execution of regular study meetings and maintenance of the manual are necessary so that the medical technologists not specializing in blood transfusion can carry out examinations confidently. An examination system is effective, but the safety of the blood transfusion treatment at night and on weekends can not be 100% secured. It is important to improve the consciousness of the clinical staff who perform blood transfusion treatment with regular training. PMID- 12652693 TI - [Computer management to avoid errors in transfusion]. AB - Even with the great progress in blood safety, such as the nucleic acid amplification test(NAT) for HBV, HCV and HIV, ABO-mismatched transfusion still occurs in the 21st century. Computer crossmatch between the patient's blood type already registered on computer, the patient's blood type as examined just before transfusion and the blood type labeled on blood bags all help avoid misunderstanding of blood type at shipping. Computerized management of the transfusion services database is also effective in promoting appropriate transfusion or autologous transfusion. Since mistakes can occur anywhere in the system, the Japanese Society of Blood Transfusion(JSBT) made a poster showing six steps where errors were most likely. They then distributed a questionnaire about mismatched transfusion to 777 major hospitals and made it clear that the main cause of mistakes was confirmation errors at bedside before transfusion. In response to the findings, JSBT created a manual for performing exactly matched and safe transfusions, including a manual for transfusion in emergencies, as well as first-line treatment procedures in case of major mismatched transfusion. In order to avoid mistakes at bedside, a barcode matching system between the patient's wristband and blood bag is ideal. It will also be useful for comprehensive risk management in modern medicine. PMID- 12652694 TI - [Diagnostic tests approved by Ministry of Health and Welfare (December 2002)]. PMID- 12652695 TI - [Endoscopic therapy for digestive system diseases]. PMID- 12652696 TI - [Endoscopic mucosal resection for superficial carcinoma of the esophagus]. PMID- 12652698 TI - [Endoscopic therapy for stomach neoplasms]. PMID- 12652697 TI - [Trends in widening application of endoscopic therapy for early-stage esophageal neoplasms]. PMID- 12652699 TI - [Trends in widening application of gastroscopic mucosal resection for early-stage stomach neoplasms]. PMID- 12652700 TI - [Endoscopic mucosal resection for early-stage colonic neoplasms]. PMID- 12652701 TI - [Trends in widening application of colonoscopic therapy for early-stage colonic neoplasms]. PMID- 12652703 TI - [Present status of laparoscopic surgery for gastrointestinal neoplasms]. PMID- 12652702 TI - [Complications of early-stage digestive system neoplasms treated with endoscopic therapy and the prognosis ]. PMID- 12652704 TI - [Endoscopic therapy for esophageal and gastric varices]. PMID- 12652705 TI - [Complications of esophageal and gastric varices treated with endoscopic therapy and the prognosis ]. PMID- 12652706 TI - [Endoscopic hemostasis for bleeding peptic ulcers]. PMID- 12652707 TI - [Laparoscopic cholecystectomy for cholelithiasis]. PMID- 12652708 TI - [Endoscopic sphincterotomy and endoscopic papillary balloon dilation therapy for choledocholithiasis]. PMID- 12652709 TI - [Endoscopic stent insertion for malignant esophageal stenosis]. PMID- 12652711 TI - [Endoscopic stent insertion for decompression of pancreatic duct stenosis]. PMID- 12652710 TI - [Endoscopic drainage stent insertion for malignant biliary obstruction]. PMID- 12652712 TI - [Recent progress on endoscopic therapy for digestive system diseases (discussion)]. PMID- 12652714 TI - [Cellulitis, sepsis, acute renal failure and hemolytic anemia with McLeod blood group phenotype]. PMID- 12652713 TI - [Hyperintense hippocampus on diffusion-weighted MR images in a case of insulinoma with persistent memory disturbance]. PMID- 12652715 TI - [Clarithromycin-induced torsades de pointes]. PMID- 12652716 TI - [Interstitial pneumonia associated with pyoderma gangrenosum successfully treated with cyclosporine A]. PMID- 12652717 TI - [Regenerative medicine for kidney: Bone marrow stem cell transplantation for glomerulonephritis]. PMID- 12652718 TI - [Therapy for hypertension: EBM and tailor-made medicine]. PMID- 12652719 TI - [Epidemiological study of hypertension in Japan]. PMID- 12652720 TI - [Diagnostic criteria for hypertension in view of evidence-based medicine]. PMID- 12652721 TI - [Reference standards for home blood pressure measurement]. PMID- 12652722 TI - [Diagnostic criteria for white-coat hypertension]. PMID- 12652723 TI - [Occurrence and diagnosis of primary aldosteronism and its therapy]. PMID- 12652724 TI - [Occurrence and diagnosis of renovascular hypertension and its therapy]. PMID- 12652725 TI - [Therapy for complication-free hypertension]. PMID- 12652726 TI - [Therapy for juvenile hypertension]. PMID- 12652727 TI - [Therapy for aged patients with hypertension]. PMID- 12652728 TI - [Therapy for hypertension associated with diabetes mellitus]. PMID- 12652729 TI - [Therapy for hypertension associated with ischemic heart disease]. PMID- 12652730 TI - [Therapy for hypertension associated with heart failure]. PMID- 12652731 TI - [Therapy for hypertension associated with kidney disease]. PMID- 12652732 TI - [Drug therapy for hypertension associated with stroke--special reference to the prevention of recurrence]. PMID- 12652733 TI - [Possibility of withdrawal of antihypertensive agents]. PMID- 12652734 TI - [Therapy for hypertension--evidence based medicine (discussion)]. PMID- 12652735 TI - [Churg-Strauss syndrome with neurogenic bladder and good recovery after steroid therapy]. PMID- 12652736 TI - [Chronic hypersensitivity pneumonitis complicated by recurrent pneumothorax]. PMID- 12652737 TI - [Primary amenorrhea and severe osteoporosis]. PMID- 12652738 TI - [Intractable hiccups treated with mexiletine hydrochloride]. PMID- 12652740 TI - [Cofactor diseases]. PMID- 12652739 TI - [Molecular targetting therapy for non-small-cell lung neoplasms with EGF-R inhibitors]. PMID- 12652741 TI - [New theory on gene regulation: mechanism of formation of chromosome functional and boundary regions]. PMID- 12652743 TI - [New perspectives on genetically engineered mice in post-genomic era]. PMID- 12652742 TI - [Analysis of hematopoietic differentiation using ES cell in vitro differentiation induction system]. PMID- 12652744 TI - [mRNA degradation in Escherichia coli]. PMID- 12652746 TI - [The breeding based on genome-wide selection]. PMID- 12652745 TI - [Sulfoglycolipids essential for the function of brain and nervous system]. PMID- 12652747 TI - [Organs formation and their transplantations in early vertebrate development]. PMID- 12652749 TI - [Standardization of microarray experiment data]. PMID- 12652750 TI - [General internal medicine]. PMID- 12652752 TI - [Geriatric medicine]. PMID- 12652751 TI - [Psychosomatic medicine]. PMID- 12652753 TI - [Preventive medicine (multiphasic screening)]. PMID- 12652754 TI - [Emergency treatment and emergency medicine]. PMID- 12652755 TI - [Medical education]. PMID- 12652757 TI - [Terminal care]. PMID- 12652756 TI - [Medical informatics]. PMID- 12652758 TI - [Medical oncology in Japan]. PMID- 12652759 TI - [Multiphasic health screening]. PMID- 12652760 TI - [Totally automated hospital laboratory transport system]. PMID- 12652761 TI - [Rural medicine and the medical care of the aged]. PMID- 12652762 TI - [Psychosomatic medicine]. PMID- 12652763 TI - [Imaging diagnosis (axial transverse tomography)]. PMID- 12652764 TI - [Clinical laboratory tests]. PMID- 12652765 TI - [Radiologic diagnosis and nuclear medicine]. PMID- 12652766 TI - [Endoscopic technics (with special reference to their therapeutic application)]. PMID- 12652768 TI - [Diagnostic pathology]. PMID- 12652767 TI - [Clinical pharmacology]. PMID- 12652769 TI - [Transition in drug designing]. PMID- 12652770 TI - [Changes in diagnostic instrument and devices in the last 100 years: discussion]. PMID- 12652771 TI - [Therapeutic guideline review: 1. Geriatrics]. PMID- 12652772 TI - [Therapeutic guideline review. 2. Psychosomatic medicine]. PMID- 12652773 TI - [Resuscitation and emergency care: "Guideline 2000"]. PMID- 12652774 TI - [Therapeutic guideline review: 4. Clinical tests]. PMID- 12652775 TI - [Therapeutic guideline review: 5. Palliative care (WHO)]. PMID- 12652776 TI - [Contribution to the study of Minamata Disease by Japanese researchers]. PMID- 12652777 TI - [Pneumatosis cystoides intestinalis in overlap syndrome manifested by dermatomyositis and scleroderma]. PMID- 12652778 TI - [IgA nephropathy in the patient with systemic lupus erythematosus in remission]. PMID- 12652779 TI - [Autoimmune pancreatitis with atypical ERP image and Eosinophilia]. PMID- 12652780 TI - [Munchausen syndrome]. PMID- 12652781 TI - [Pulmonary lymphangioleiomyomatosis]. PMID- 12652782 TI - [Acquired coagulation abnormality due to factor V inhibitor]. PMID- 12652783 TI - [The development and clinical implication of genetic tests]. AB - Genetic testing is not performed as often as other clinical laboratory tests currently available. However, physicians will in the future be eager to analyze and diagnose the diseases at a genetic level, since the DNA sequence analysis of the human genome has been completed. Moreover, information on single nucleotide polymorphisms in disease susceptibility and drug responsiveness is increasing, and genetic testing based on this information will be utilized for the prevention of disease and order-made medicine. Hence, the demand for genetic tests will increase in the near future. In this review, the development and clinical implication of genetic tests will be discussed. PMID- 12652784 TI - [The ethical implications, guidelines, and standardization of genetic tests]. AB - Laboratory advances in molecular genetics have resulted in numerous clinical applications for DNA analysis. Genetic tests can contribute a great deal of information to clinical diagnosis. These genetic tests including PCR, fluorescence detection, real-time PCR, and automated sequencing have developed into both simple and time-consuming laboratory techniques. Currently, DNA diagnosis is not used routinely because of cost, complexity, and resources. We have focused on the ethical implications and proper standardization in DNA diagnosis. This review is to discuss a guide to proceed genetic tests for their efficiency, accessibility and quality in laboratory. PMID- 12652785 TI - [How to make the best use of the Internet]. AB - Presently, searching for genetic information successfully requires using the internet. Because the internet world has different common sense than the publication world, those who are going to use the internet should know its characteristics well. First, because there is an overwhelming number of web sites that continue to change, proper techniques for making an efficient search for valuable information are necessary. Second, the contents of web sites need validation. Third, possible harms that can come with the internet should be avoided beforehand, and so on. Recent rapid progress in information technology will soon bring the internet to every field of medical practice and the above mentioned know-how will become minimum requirements for every clinician. PMID- 12652786 TI - [Recent advances in molecular diagnostic tests]. AB - Recent advances in molecular biotechnologies have facilitated laboratory use of molecular diagnostic tests. Automated systems for amplification and detection, and lately for extraction, of nucleic acid sequences have been developed. The automated systems have allowed improvement of not only assay efficiency but also quality control of the tests. The information on the genome sequence from the human genome project has been studied to elucidate functions of genes and proteins and the clinical significance of nucleic acid sequences with post genomics such as expression profiling using DNA microarray, proteomics, and single nucleotide polymorphism analysis, in conjunction with bioinformatics. Now the challenge is the development and application of these new technologies as clinical diagnostic tools. Design of a diagnostic array or panel must be developed with defined sequences based on interpretation of a huge quantity of experimental data to meet clinical demands. There is a need for development of generally available instruments that are inexpensive, practical and more importantly reproducible and reliable. PMID- 12652787 TI - [Molecular diagnosis of hepatitis B virus and hepatitis C virus infection]. AB - Detection of virus genomes and their mutants by molecular technology is essential in clinical practice for patients with hepatitis B virus(HBV) and hepatitis C virus(HCV) related liver diseases. We here briefly summarize genome structures of HBV-DNA and HCV-RNA. Also, we describe representative methods to detect and quantify HBV-DNA and HCV-RNA. PMID- 12652789 TI - [MRSA infection]. AB - Molecular diagnosis of MRSA infection detecting mecA, spa gene and toxin gene by PCR including genotyping methods for MRSA nosocomial outbreak investigations are described. Restriction length polymorphism using pulsed-field gel electrophoresis(PFGE-RFLPs) with SmaI restriction enzyme is widely used for elucidating the infection route of MRSA. In Japan, MRSA is the major pathogen for nosocomial infections. These laboratory findings play an important role in infection control. PMID- 12652788 TI - [Diagnosed tuberculosis using specific DNA probe hybridization methods]. AB - In Japan, reported cases of tuberculosis had declined nearly every year until 1995. However, in 1997 newly recorded cases began increasing for the first time in more than 38 years. Recent studies using DNA fingerprinting show that person- to person transmission may account for as many as one-third of new cases of tuberculosis in citizen populations. Nucleic acid hybridization methods using specific DNA probes can specifically identify M. tuberculosis and other mycobacterial species. Rapid nucleic acid amplification techniques such as polymerase chain reaction methods allow direct identification of M. tuberculosis in clinical specimens. Is 6110 has been exploited extensively as a clonal marker in molecular epidemiology studies of tuberculosis. The emergence of resistance to antituberculosis drugs is a relevant matter worldwide. A recent genotypic method allows earlier detection of RFP-resistant and INH-resistant stains using probes for mutation in rpoB and in katG. PMID- 12652790 TI - [Human immunodeficiency virus infection]. AB - Decisions regarding initiation or changes in antiretroviral therapy in patients with HIV-1 infection should utilize plasma HIV RNA(viral load) assay and drug resistance assay. These laboratory tests give the physician important information about the virologic and immunologic status of the patient and risk of disease progression to AIDS. Testing for drug resistance to antiretroviral drugs may help to maximize the benefits of antiretroviral therapy. Genotyping assays detect drug resistance mutations and phenotyping assays measure the ability of viruses to grow in various concentrations of antiviral drugs. The presence of drug resistance and failure of antiretroviral therapy should be identified by these two assays. PMID- 12652791 TI - [The infections by Chlamydia trachomatis and Neisseria gonorrhoeae]. AB - Among sexually transmitted diseases, Chlamydia trachomatis and Neisseria gonorrhoeae infections are found most frequently. Recently, their incidence in youths has increased. The rate of infection by human immunodeficiency virus is increased in patients who have been infected by Chlamydia trachomatis. In view of this, early detection and treatment of Chlamydia trachomatis infection appear to be important. Asymptomatic infections by Chlamydia trachomatis and Neisseria gonorrhoeae are not rare among gynecological patients. However, in patients infected by both of these pathogens it is difficult to detect microbial antigens using specific antibodies. Therefore, patients suspected of infection by these pathogens should receive precise genetic testing for Chlamydia trachomatis and Neisseria gonorrhoeae, and the results of such tests should be evaluated carefully. PMID- 12652792 TI - [Vancomycin resistant enterococcal infections and detection of their resistance genes]. AB - Vancomycin-resistant enterococci(VRE) is a troublesome pathogens causing nosocomial infections worldwide. VRE infections or colonization occur mainly in immuno-compromised patients and in the course of antimicrobial treatment for other nosocomial infectious diseases. VRE are classified into 6 types by their resistance genes and of these types, both VanA and VanB type VRE are significantly important in clinical situations. Susceptibility tests for VCM and TEIC are often useful in the phenotypic screening for differentiating VanA from VanB type VRE. However, detection of the resistance genes using PCR methods is easy and essential for the conclusive identification of VRE. PMID- 12652793 TI - [Cytomegalovirus infection]. AB - By application of PCR technology, Cytomegalovirus(CMV) DNA can be detected from clinical specimens, rapidly and with high sensitivity. PCR-based assays can detect CMV DNA from blood even prior to the onset of clinical symptoms, so qualitative PCR assays are used as an indicator for preemptive therapy in transplant recipients. But it is not always helpful in CMV infection because it cannot distinguish latent from active infection, meaning lower positive predictive value for clinical disease. In recent years, quantitative PCR assays are increasingly being utilized in the clinical setting to (i) identify individuals at risk of developing CMV disease(determination of clinical threshold), (ii) monitor viral response to antiviral treatment, (iii) predict individuals at risk of clinical relapse, and (iv) serve as an early indicator of antiviral resistance. PMID- 12652794 TI - [Bacillus anthracis, prions, Coxiella burnetii]. AB - The detection kits for Bacillus anthracis, an isoform of host prion and Coxiella burnetii, using genetic technology are still not generalized. For B. anthracis target genes for detection would be genes of toxins. Although it is difficult to detect prions related to prion diseases before death, some mutants of prion gene in leukocytes and 14-3-3 proteins in cerebrospinal fluid are detectable in living patients suffering from Creutzfeldt-Jakob disease. The gene of superoxide dismutase in Coxiella burnetii is a useful target for detecting this Rickettsiae. PMID- 12652795 TI - [Genetic tests for leukemia]. AB - The diagnosis of leukemia is mainly based on chromosome analysis and genetic testing. The analysis of molecular genetics associated with chromosome abnormalities has contributed to new classifications of leukemia. The minimal residual disease(MRD) in its currently accepted application refers to low-level disease detected by quantitative PCR techniques in clinical situations. It is anticipated that efforts in characterization of molecular genetics in leukemia will ultimately translate into better clinical outcomes for patients. PMID- 12652796 TI - [Gene level diagnosis in malignant lymphoma]. AB - Malignant lymphoma is a diverse group of clonal proliferative disorders derived from immature and mature lymphoid cells. Each subtype of lymphoma is defined according to a combination of morphology, immunophenotype, genetic features and clinical syndromes. Accordingly, based on our practical experience in the laboratory-oncologic point of view, the role, clinical significance, applications and laboratory issues for testing or interpretation of gene level examination in practical oncology of lymphoma are reviewed. PMID- 12652797 TI - [Genetic background of Japanese thrombophilia]. AB - Coagulation Factor V Leiden has not been detected in Japanese patients suffering from thromboses. The genetic background of Japanese thrombotic patients has never been systematically examined. We have performed a systematic investigation to ascertain genetic risk factors for thromboses in the Japanese population. According to our results, we recommend here useful gene analyses for Japanese thrombophilia. PMID- 12652798 TI - [Platelet disorders]. AB - Platelets play important roles in thrombosis and hemostasis, and their abnormalities lead to hemostatic disorders or thrombotic diseases. Due to genetic technique development, the study of genetic factors related to the hereditary platelet dysfunction, and the evaluation of gene polymorphisms have been reported recently. In this article, we will focus mainly on hereditary platelet disorders and genetic variations of platelet glycoproteins involved in the pathogenesis of thrombotic diseases. PMID- 12652799 TI - [Familial (hereditary) cancer]. AB - When cancer patients accumulated in one pedigree, the cancer is named as familial cancer. Especially when there is strong genetic background, It is named as hereditary cancer. Most of them are caused by genetic mutation in a gene related to cancer susceptibility. The gene is classified among suppressor oncogenes, oncogenes or DNA repair genes. Although hereditary cancer is uncommon, genetic testing is very useful to confirm hereditary cancer and to diagnose it in the presymptomatic stage. On the other hand, it poses ethical problems, therefore a strategy including genetic counseling is essential. PMID- 12652800 TI - [Oncogene and tumor suppressor gene]. AB - Various gene information is being accumulated about the mechanisms of carcinogenesis and the characteristics of cancer cells with progress in molecular biology. However, when two or more genetic abnormalities are found in cancer, it is not easy to apply the analysis of oncogenes or tumor suppressor genes to routine laboratory examination. Further study is necessary. PMID- 12652801 TI - [Telomerase]. AB - Telomerase, an intranuclear enzyme, binds to telomeres at chromosome ends, helps to elongate telomere length and is closely linked to attainment of cellular immortality, a step in carcinogenesis. Since telomerase activity has been detected in several human tumors and hematopoietic malignancies, this enzyme is thought to be a useful marker for cancer diagnosis. Among the components of telomerase, human telomerase catalytic subunit(hTERT) is the most important regulatory factor for telomerase activity. Therefore, quantitation of hTERT mRNA expression may be a new approach to measuring telomerase activity. PMID- 12652802 TI - [Glycosyltransferase genes as tumor marker]. AB - Core 2 beta 1,6-N-acetylglucosaminyltransferase(C2GnT) and alpha 1,4-N acetylglucosaminyltransferase are glycosyltransferases involved in the biosynthesis of mucin type glycoprotein(O-glycan). The transcripts of C2GnT, which forms core 2-branched O-glycan(Gal beta 1-->3 (GlcNAc beta 1-->6)GalNAc alpha-->Ser/Thr), were detected approximately in 2/3 cases of patients with colorectal or lung cancers. Then, carcinoma cells expressing C2GnT mRNA were shown to significantly progress compared with those lacking the C2GnT mRNA, indicating an important role of the core 2-branched O-glycan in tumor progression. On the other hand, gland mucous cell-type mucin secreted from the normal gastric mucosa characteristically contains GlcNAc alpha 1-->4Gal beta-->R structure, and the alpha 4GnT is critical for the biosynthesis of this unique glycan. This enzyme is also detected in gastric cancer cells but not in mononuclear cell fraction of the peripheral blood. Thus, the quantitative RT-PCR method targeted to alpha 4GnT mRNA will be useful for the detection of circulating gastric cancer cells in the peripheral blood. PMID- 12652803 TI - [Cytokine gene]. AB - The most important aspect of cytokine gene expression in cancer is paraneoplastic syndrome. Paraneoplastic syndrome presents symptoms and signs characteristic of substances that tumors produce, some of which have been revealed to be cytokine. Detection of cytokine gene expression in paraneoplastic syndrome is important not only for diagnosis but for monitoring of anti-cytokine therapy that is expected to be rapidly developed. Proper diagnosis and treatment of paraneoplastic syndrome are also important for improving the QOL of cancer patients. In this context, recent advancement of anti-cytokine monoclonal antibody therapy is expected to aid treatment. PMID- 12652804 TI - [Aspiration biopsy RNA diagnosis]. AB - Molecular-based diagnosis of carcinomas can be more easily established by utilizing specific mRNAs that are restrictively expressed in cancer tissues. Recently a new method of preoperative molecular-based diagnosis of thyroid carcinomas was established. This technique, Aspiration Biopsy RNA Diagnosis, ABRD, allows us to perform preoperative RNA analyses by extracting RNAs from tumor cells obtained by fine needle aspiration biopsies(FNABs). By ABRD detecting of oncofetal fibronectin or calcitonin mRNA in FNABs, papillary, anaplastic and medullary carcinomas are accurately diagnosed preoperatively. Further, by real time monitoring RT-PCR, a fully automated system was established. PMID- 12652805 TI - [Genetic tests in the neurological disorders]. AB - Most of the genetic tests for infectious diseases have few ethical problems and directly link to treatments. On the other hand, there are many problems in performing genetic tests for hereditary neurological disorders without essential treatments. Patients who undergo a genetic test usually get few benefits when they have a positive result. Moreover, the results may influence on their family members' daily lives. Therefore, we should be careful in genetic testing of the hereditary neurological disorders, and perform sufficient informed consent, genetic counseling of the patients and their family members, and precise standardization. PMID- 12652806 TI - [Endocrine and metabolic diseases]. AB - Multiple gene mutation inducing common metabolic diseases(diabetes mellitus and hyperlipidemia) and single gene mutation inducing congenital metabolic diseases or inherited endocrine disease are the main targets of DNA diagnosis. We review our current understanding of DNA diagnosis of representative inherited endocrine and metabolic diseases. PMID- 12652807 TI - [Genetic diagnosis for inherited lung disorders]. AB - Technological advances in molecular biology, as well as in genetic engineering, have created a revolutionary movement in both clinical and diagnostic methodologies. With progress in human genome analysis, the genes responsible for various diseases have been analyzed at a rapid pace. Research involving respiratory disease is no exception. Emphasis in this area has shifted from respiratory physiology to molecular biology. One example is cystic fibrosis(CF). However, CF is relatively rare in Japan. Furthermore, the search for universal genes that trigger bronchial asthma has not been successful. Genetic testing for respiratory disease has not been conducted as a component of general, routine clinical tests in Japan, due to the aforementioned reasons. Therefore, I would like to take this opportunity to introduce genetic respiratory diseases, the present state of genetic testing and prospects for the future. PMID- 12652808 TI - [Inherited renal and vascular diseases]. AB - The advances in molecular biology during the last decade have revealed the genes responsible for inherited renal and vascular diseases and their mutations. These genes code structure proteins, receptors, channels, transporters, and signal proteins, which are specifically expressed in the renal and vascular systems. Other genes code modulators of the system. These findings have provided the understanding of molecular mechanisms of hereditary disorders and may open the way for genetic diagnosis of inherited renal and vascular diseases in the field of laboratory medicine. PMID- 12652809 TI - [Genetic tests in immune system-related diseases]. AB - Recent progress in molecular medicine has revealed genes responsible for numerous instances of immunodeficiency syndrome. Such disorders have been diagnosed mainly by functional tests of both T and B lymphocytes. Genetic tests are effective in cases difficult to diagnose as well as in the detection of gene carriers. Currently, studies are underway to determine if SNP significantly impacts polygenic immunosystem-related diseases, and whether its effects influence disorders secondary to such diseases. In some diseases analysis of HLA aids in diagnosis. PMID- 12652810 TI - [Roles of molecular analysis in hematopoietic stem cell transplantation]. AB - Recent advances in molecular analysis by polymerase chain reaction(PCR) have taken a prominent position an essential part in regeneration medicine. In hematopoietic stem cell transplantation, HLA(human leukocyte antigen) typing, assessment of graft viability/rejection (chimerism analysis), and evaluation of minimal residual disease are significant for treatment strategy. Molecular analysis will play a more important role in the diversification of transplantation methods in the future of tailor-made medicines. PMID- 12652811 TI - [Gene therapy]. AB - Gene therapy is a method for treating hereditary disease at the gene level. In this paper, I show recent gene therapy approaches for cancer, hemophilia and arteriosclerosis obliterans, and gene medical supply including antisense and decoys are also introduced. With the rapid advances in gene therapy research, the case of dying gene therapy using the adenovirus vector was reported due to DIC. Establishment of the safety in the gene therapy and solution of the ethics problems are also important issues. PMID- 12652812 TI - Emphysematous changes are caused by degradation of type III collagen in transgenic mice expressing MMP-1. AB - Disruption of the extracellular matrix is believed to play an important role in the pathogenesis of emphysema. Prior studies have demonstrated that transgenic mice expressing the human tissue collagenase, matrix metalloproteinase 1 (MMP-1), develop emphysema. MMP-1 is a protease with substrate specificity for fibrillar collagen. Type I and III collagens, which are the most abundant proteins within the lungs, are the primary substrates for MMP-1. To assess if type I collagen was indeed the site of action for MMP-1 in these transgenic mice, hybrid mice were generated by crossing the MMP-1 transgenic mice with mice that had degradation resistant type I collagen. The hybrid mice demonstrated an identical emphysematous phenotype as the MMP-1 transgenic mice, indicating that the degradation of type I collagen was not essential to the development of emphysema in these mice. Immunohistochemical studies in control mice demonstrated that collagen fibers in the alveolar walls and ducts of the normal mouse lungs consist mainly of type III collagen. In the transgenic and hybrid mice, the emphysematous changes, which developed, were associated with a marked decrease in type III collagen in these alveolar structures. These results indicate that MMP-1 generated the emphysematous phenotype via the degradative effect on type III collagen, which is a vital structural element of the alveolar walls. This is the first study to show that a matrix metalloproteinase may cause emphysema via its effects on a specific collagen subtype. As such, it should provide important insight into the mechanisms of this disease in humans. PMID- 12652813 TI - Cigarette smoke decreases inducible nitric oxide synthase in lung epithelial cells. AB - Cigarette smoking has been associated with decreased exhaled nitric oxide (NO). To investigate the mechanism of this decrease, the effects of a cigarette smoke extract were evaluated a murine lung epithelial cell line (LA-4), a human lung epithelial cell line (A549), and primary cultures of human lung epithelial cells induced to produce NO by cytokines. NO production was evaluated by measuring nitrite, a stable end product of NO, in cell culture supernatant fluids. Cigarette smoke extract caused a reduction in the cytokine-induced nitrite concentrations in the culture supernatant fluids from all 3 cell types (P < .01, all comparisons). To further investigate these observations, immunohistochemistry demonstrated a decrease in cytokine-induced inducible NO synthase (iNOS) protein expression and iNOS mRNA after cigarette smoke extract exposure in LA-4 cells. However, iNOS mRNA half-life was not altered by the smoke extract, suggesting that the smoke extract decreased NO by decreasing iNOS mRNA transcription. These findings demonstrate that cigarette smoke extract decreases iNOS expression and NO production from lung epithelial cells. PMID- 12652814 TI - Response of human alveolar macrophages to ultrafine, fine, and coarse urban air pollution particles. AB - In the lower airways, macrophages are important regulators of inflammation and indispensable in their antimicrobial activities. Thus, air pollution particles, which modulate airway macrophage host defenses may, in susceptible individuals, increase severity of inflammatory and infectious disease. In the present study, size fractionated, ultrafine (UF), fine (PM0.1-2.5), and coarse (PM2.5-10) particulate matter (PM) were collected from 2 urban sites in the Netherlands, and were compared for effects on human alveolar macrophages (AM). Inflammatory cytokine production, phagocytosis, and expression of phagocyte receptor CD11b were assessed in particle-exposed AM. Interleukin (IL)-6 levels induced by PM2.5 10 (20411 pg/mL) were > 10-fold higher than induced by PM0.1-2.5 (1781 pg/mL). Levels induced by PM0.1-2.5 were 2- to 3-fold higher than induced by UF (770 pg/mL) when cells were exposed to the same particle mass. Cytokine induction by the PM was inhibited by antibody to CD14 and required the presence of serum for optimal stimulation, implying that bacterial products or endotoxin were stimulatory moieties in both coarse and fine particulate matter. Phagocytosis of opsonized yeast was inhibited by coarse more than by fine PM, as was yeast induced oxidative burst. Coarse particles decreased CD11b expression more than fine PM. The UF did not affect these functions. Taken together, these results suggest that PM recognition by human AM involves receptors evolved to recognize microbial cell structures, and that microbial products preferentially found in the coarse particle fraction of PM may be involved in inflammatory events and decreased pulmonary defenses associated with exposure to pollution particles. PMID- 12652815 TI - Imaging lung inflammation in a murine model of Pseudomonas infection: a positron emission tomography study. AB - We tested the hypothesis that the uptake of [18F]fluorodeoxyglucose (FDG), as measured by positron emission tomography (PET) imaging, would correlate with inflammation caused by increasing doses of instilled Pseudomonas aeruginosa (PA) into the lungs of mice. PA-laden agarose beads were instilled via the trachea into 1 lung of each mouse (dose range 0.5-15 x 10(4) CFU) and imaging was performed 3 days later (at the peak of the inflammatory response). Lung uptake of [18F]FDG correlated significantly with the dose of bacteria instilled in mice infected with the M57-15 strain of PA (n = 18) (r2 = .62), but not in mice infected with the PA01 strain (n = 20). The overall lung uptake of [18F]FDG was higher in mice infected with the M57-15 strain than in those infected with the PA01 strain (P < .05). Total white blood cell concentrations in bronchoalveolar lavage were also higher in the M57-15-infected mice. We conclude that PET imaging can detect and quantify differences in host inflammatory response to 2 different strains of PA. The combination of PET imaging with murine models should be a useful new tool to study neutrophil trafficking and kinetics in lung inflammation. PMID- 12652816 TI - [Restoring sleep patients. Snoring as nocturnal nerve irritant]. PMID- 12652817 TI - [MMW questions colleagues about prescriptions. How to manage children with otitis media? (interview by Thomas Meissner)]. PMID- 12652818 TI - [Itching, pain or bleeding: your patient will always blame "hemorrhoids"]. PMID- 12652819 TI - [Safer contraception? The "pill"--also for the woman after age 35?]. PMID- 12652820 TI - [Drug for Gaucher disease also works as sperm inhibitor. Is the "pill" for the man on its way?]. PMID- 12652821 TI - [Unfamiliar with such high survival rates in tumors. Are we doing everything wrong here?]. PMID- 12652822 TI - [Vitamins, statins, ACE inhibitors. What really prevents stroke?]. PMID- 12652823 TI - [Transient cerebral ischemia--precursors of stroke. Can warning signs be recognized?]. AB - Transient ischemic attacks (TIA) are neurological deficits of sudden onset and equally sudden reversal. They are considered to be precursors of stroke that (still) have a good prognosis. Since the symptoms have usually disappeared by the time of the first examination the physician must rely on the information provided by the patient or relatives for the medical history. Symptoms to look for include disordered speech, aphasia, vertigo, unilateral arm paralysis, crooked mouth. When a TIA is suspected, the following examinations are mandatory: neurological ultrasound, blood pressure, ECG, laboratory examinations (blood sugar, blood count, hematocrit), CT or MRI, Doppler-ultrasonography of the carotids and, where indicated, transcranial Doppler. Long-term prophylaxis with platelet antiaggregation agents or Marcumar (phenprocoumon) and appropriate changes in lifestyle are indicated for all TIA patients. When the indication is established, thrombarterectomy must be carried out within six months after the event. PMID- 12652824 TI - [Drug therapy of stroke in the stroke unit. A race against time]. AB - Ischemic stroke is usually a manifestation of arteriosclerosis in the elderly patient. The major risk factors are age and high blood pressure; prevention is particularly successful in hypertensives. All known antihypertensive agents can be applied, and ACE inhibitors and the lipid-lowering agents, in particular the statins are now well established. That said, under certain circumstances it might be of importance to pay greater attention to the other lipid fractions. It is currently estimated that only one-third of patients with a disturbance of lipid metabolism actually receives treatment. Results of treatment in the acute phase of apoplexy have been improved by the establishment of stroke units that provide the facilities for intensive and comprehensive management of stroke victims. Inhibitors of platelet aggregation, in particular aspirin, now have a permanent role in the acute and follow-up phase. For specific risk factors such as stenosis of the carotid artery or atrial flutter, special therapeutic guidelines apply. PMID- 12652825 TI - [Coins, razor blades, button-sized batteries, etc. Ingested foreign bodies--when are they a true emergency?]. PMID- 12652826 TI - [Radiosynoviorthesis in active arthrosis. Will radiotherapy eliminate joint pain?]. PMID- 12652828 TI - [Suspected tumor in an elderly patient. Night sweats and subfebrile temperatures as diagnostic clues. Reactivated pulmonary tuberculosis]. PMID- 12652827 TI - [Isolated systolic hypertension. Need for new assessment]. PMID- 12652829 TI - [A new antihypertensive drug. Successful duo now as fixed combination]. PMID- 12652830 TI - [Fluoroquinolone superior to macrolide. Longer infection free period after acute bronchitis]. PMID- 12652831 TI - [When dental extraction or surgical intervention is anticipated. Coxib decreases postoperative pain]. PMID- 12652832 TI - [80% of patients are unhappy with guidelines issued by their physician. 3 ways to motivate the diabetic patient]. PMID- 12652833 TI - Prenatal smoking and birth outcomes among Mississippi residents. AB - In this study we examined trends within Mississippi in prenatal smoking and in the effects of such smoking on birthweight, preterm delivery, and infant mortality. The study was a retrospective cohort analysis of 120,429 singleton births in 1995-1997. We found that even though prenatal smoking is decreasing overall, it is increasing among young pregnant women aged 15-19 years. The primary effect of prenatal smoking was to lower birthweight; correspondingly, the principal effect of smoking on infant death appeared to be this decreasing of birthweight. In addition, infants of mothers who smoked during pregnancy were two and one-half times as likely to die from SIDS as were infants whose mothers did not smoke. These data demonstrate the importance of strategies such as training in smoking cessation for providers of prenatal care in Mississippi and provide a foundation for future evaluations of current aggressive anti-tobacco campaigns in the state. PMID- 12652834 TI - I am following a health care professional who, I think, has a drug problem. PMID- 12652835 TI - Looking back: medical inspection of school children in 1920. PMID- 12652836 TI - The rest of your mother: the story of Eudora Welty, Ouida Keeton, and the "Legs" murder. PMID- 12652837 TI - [Epidemiology is a developing system of views]. PMID- 12652839 TI - [Effect of heliogeophysical factors on epizootic activity of a native focus of visceral leishmaniasis]. AB - Visceral leishmaniasis (VL) morbidity in the population of the south-eastern Turkmenistan and the epizootic activity of a natural focus were studied. A definite cyclic recurrence of the processes occurring in the natural focus of VL, which was associated with solar activity changes was found. The regularities found make it possible to predict the increase in the epizootic tension of a natural focus of VL in this region and the incidence of this disease among the population in 2002-2004. PMID- 12652838 TI - [Structural organization and function of parasitic systems]. AB - The structural organization and functioning of parasitic systems are considered in the context of the general theory of systems. By using the parasitic system of an epizootic process of plague as an example, the essence of the concepts of the structural organization and functioning of the systems is determined, and a scheme of a relationship of elements in the trinomial parasitic systems is presented. The mechanism of parasite-host interaction at the molecular, cellular, body's, and populational levels is considered on the basis of the theory of immune-specific wave information in an living organism. PMID- 12652840 TI - [Effect of infection rate on parasite-host relations in opisthorchiasis]. AB - The adaptation of the immunity system to recurrent helminth contamination in the natural development of opisthorchiasis accords with that of the underlying adaptation syndrome in chronic stress. The established correlation makes itself evident in the fact that in recurrent infections with Opisthorchis, the acute (a mobilization phase) and particularly chronic (a depletion phase) stages of infestation are characterized by a significant reduction in the overall and specific immunological responsiveness of the host, which is followed by the enhanced biological activity of the parasite. In recurrent infections with metacercaria at the stage of invasion (an adaptation phase), the host's organism is able to retard the biological activity of the parasite by straining the immune system against specific Opisthorchis) and heterological (sheep red blood cells):antigens. With this, Opisthorchis eggs double in number after the third contamination while those increase by 10 times after the third contamination and by 26 times in the presence of chronic infection just after the second contamination with helminths. PMID- 12652841 TI - [Comparative evaluation of efficacy of treating giardiasis with drug- or phytotherapy]. PMID- 12652843 TI - [Transfusion malaria]. PMID- 12652842 TI - [Parasitic diseases in the structure of acute intestinal illnesses]. PMID- 12652844 TI - [Recording a case of dirofilariasis in residents of Novosibirsk]. PMID- 12652845 TI - [Dirofilariasis in residents of the Ulanovsk region]. PMID- 12652846 TI - [Primary echinococcosis of the heart in pulmonary tuberculosis]. AB - The paper presents a sectional case of primary cardiac Echinococcus infection in a patient with fibrocavernous pulmonary tuberculosis. A whitish thin-wall round oval cyst, 2 cm across, was detected in the thickness of the lateral left ventricular wall at autopsy. The tinctorial properties of the cyst sheath suggested that the parasite might be viable. The internal germinal layer of its sheath showed minor filial vesicles provided with scolexes. An extensive area of tissue detritis was perifocally detectable near the cystic wall, then peripherally there was a fibrous capsule with developed plethoric vessels and polymorphically cellular inflammatory infiltrate with a predominance of lymphocytes, macrophages, eosinophilic leukocytes, and fibroblasts. Parasitic cardiac damage along with tuberculous infection of respiratory organs had played an important role in the decompensation of performance of the hypertrophic heart and referred to the unified category of the second malaise in the mixed underlying disease. PMID- 12652847 TI - [Toxocariasis in the Tula region]. AB - Serological survey of 2586 persons with suspected toxocariasis revealed Toxocara antigen titers in 644 (24.9%). Canine toxocariasis affliction was 25%, environmental Toxocara egg contamination was 3.2%. Seropositive persons were recorded in 22 of the 24 districts. A patient-carrier ratio was 1:1.6. Visceral toxocariasis was more frequently notified in children. The clinical picture was polysymptomatic with a predominance of intoxication (86%), temperature responses (49%), abdominal (37%) and pulmonary (30%) syndromes. PMID- 12652848 TI - [Forgotten parasitic diseases]. AB - The ecological flexibility of parasitic systems is observed in parasites, such as pentastomas of Linguatula serrata and Armillifer sp., cestodes of Spirometra erinacei europaei, and dirofilariae of D. repens, D. immitis, D. ursi in relation to ecological shifts in the areas and anthropological pressure. PMID- 12652849 TI - [Human fascioliasis caused by F. hepatica]. AB - The paper deals with the possibility of invasion and parasitizing of Fasciola hepatica in man. Data of observations and references are presented. PMID- 12652850 TI - [Measures for controlling the first intermediate host and larval stages of opisthorchis]. PMID- 12652851 TI - [Mosquitos (Phlebotominae) in the Russian Federation]. PMID- 12652852 TI - [Modern aspects of pathogenesis of trophic malaria]. PMID- 12652853 TI - EMTALA impact on emergency physicians. PMID- 12652854 TI - Health insurance reform. PMID- 12652855 TI - Fed up with the insurance industry, physicians strike back. PMID- 12652856 TI - Tort reform and the professional liability climate. PMID- 12652857 TI - Compliance in the medical office. Take the positive approach. AB - This article presents an overview of the primary compliance regulations impacting medical practices in Maryland. While acknowledging the impact compliance has in terms of time and cost, the focus of the article is on making compliance plans advantageous to each medical practice. The particular focus is on minimizing the negative consequences of government regulations and implementing compliance plans that make sense for your office. PMID- 12652858 TI - Disparities in health care treatment. PMID- 12652859 TI - WAITT. We're all in this together, aren't we? PMID- 12652860 TI - Impacting stroke in Maryland. PMID- 12652861 TI - Smallpox update. PMID- 12652862 TI - Art, artists, and arthritis. PMID- 12652863 TI - [Streptococcus pneumoniae strains resistance to macrolide, lincosamide, streptogramin, oxazolidinone and ketolide]. AB - Over the last two decades, antimicrobial resistance in Streptococcus pneumoniae has been detected at an increasing rate. There have been several reports of increasing rate of macrolide resistance among the penicillin-resistant S. pneumoniae strains. The rapid evolution of such resistance necessitated new antimicrobials, such as streptogramins, oxazolidinones and ketolides, against beta-lactam and macrolide resistant pneumococci. This study was undertaken to determine the prevalence of macrolide resistance in pneumococci isolated in a Turkish University hospital and also to determine the activity of these newly developed agents against pneumococci. For that purpose a total of 264 pneumococci, isolated from clinical specimens, were tested for susceptibility against penicillin, erythromycin, clarithromycin, clindamycin, quinupristin/dalfopristin, linezolid and telithromycin by agar dilution method. Penicillin resistance and intermediate penicillin resistance was found in 7.6% and 40% of isolates respectively, while resistance rates of the tested isolates against erythromycin, clarithromycin and clindamycin were as 15.9%, 13.6% and 13.6%, respectively. Resistance to macrolides and clindamycin was higher among the penicillin-resistant isolates. No resistance was detected against quinupristin/dalfopristin, linezolid and telithromycin, except for four strains which had minimal inhibitory concentration (MIC) values of intermediately susceptible category to quinupristin/dalfopristin. These data indicate the presence of an important percentage of macrolide resistant pneumococci in our hospital. It can also be concluded that streptogramins, oxazolidinones and ketolides may be used as good alternatives especially in case of infections due to macrolide resistant pneumococci. PMID- 12652864 TI - [Comparison of routine sensitivity tests and mecA gene analysis for determination of methicillin resistance in Staphylococcus aureus isolates]. AB - Currently, oxacillin disk diffusion test is the most frequently employed method to detect the methicillin resistance of Staphylococcus aureus isolates. However, due to some of the test conditions, errors may occur during the detection of heteroresistant bacteria. It is now widely accepted that lower incubation temperatures (< or = 35 degrees C) and media with a NaCl concentration of 2-4%, could facilitate detection. In our study, methicillin (oxacillin) susceptibilities of 125 S. aureus isolates were determined by the disk diffusion and microdilution tests as defined by National Committee for Clinical Laboratory Standards (NCCLS), and the results were compared with those of mecA gene analysis. In the routine susceptibility tests, 75 isolates were found to be methicillin resistant (MRSA), whereas 50 were found to be susceptible (MSSA). Various induction tests were performed to investigate the heterogeneous resistance among methicillin-susceptible isolates. These induction tests showed that the MIC values of seven isolates reached to the resistant levels, therefore these isolates should be accepted as "borderline oxacillin resistant S. aureus" isolates lacking the mecA gene. The susceptibility tests and mecA gene analysis of the remaining isolates yielded compatible results. In conclusion, susceptibility tests, when performed according to NCCLS recommendations, are found to be reliable and decisive, for the detection of methicillin resistance of S. aureus. PMID- 12652865 TI - [In vitro sensitivity of methicillin-resistant Staphylococcus aureus to fusidic acid and trimethoprim-sulfamethoxazole]. AB - In this study, a total of 225 methicillin resistant Staphylococcus aureus strains isolated from various clinical specimens (114 wounds, 27 blood, 16 of each catheter and sputum, 14 throat swabs, 10 tracheal aspirates, 7 of each urine and drain materials, 6 pleural fluids, 5 graft materials, 2 cerebrospinal fluids, 1 ascitis fluid) of hospitalized patients, were tested in-vitro for their susceptibilities to fusidic acid and trimethoprim-sulfamethoxazole (TMP-SMX) by disk diffusion method. Two-hundred-eighteen (96.9%) of isolates were found susceptible to both fusidic acid and TMP-SMX, 4 (1.8%) were resistant to both of the agents (3 isolates from wound, 1 from sputum), 2 (0.9%) (both of them were wound isolates) were resistant only to fusidic acid, and 1 (0.4%) (throat isolate) was resistant only to TMP-SMX. PMID- 12652866 TI - [Penicillin tolerance in group A beta-hemolytic streptococci isolated from throat cultures of children with tonsillopharyngitis]. AB - The most common bacterial cause of tonsillopharyngitis is group A beta-hemolytic streptococci (GABHS) and penicillin is the drug of choice for the treatment of these infections. However, a disturbing trend toward penicillin failure has developed in some communities. It has been proposed that penicillin tolerance is a possible explanation for this failure. In this study, penicillin tolerance of GABHS strains isolated from 263 children with tonsillopharyngitis, was investigated between December 2000-March 2001. As a result, none of the isolated strains was found to be penicillin resistant or penicillin tolerant according to minimal inhibitory concentration and minimal bactericidal concentration values for penicillin G, determined by microdilution method. PMID- 12652867 TI - [Evaluation of western blot methods for serologic diagnosis of Helicobacter pylori infections]. AB - Recent studies have demonstrated that immunoblot method is more sensitive and specific than enzyme immunoassays (EIA) to diagnose Helicobacter pylori infections, and it also permits the detection of antibody profiles against different bacterial antigens including the virulence factors. The aim of this study was to evaluate the diagnostic value of Western Blot (WB) method in the serological diagnosis of H. pylori infections. For this purpose, H. pylori IgG and IgA antibodies have been searched qualitatively by a commercial WB test (Euroimmun GmbH, Germany), in the sera of 23 patients (9 female, 14 male) ages between 6-70 years old (13 children, 10 adults), who were diagnosed as H. pylori infection with the positive results of at least three of the conventional methods (culture and polymerase chain reaction of endoscopic biopsy specimens, 13C urea breath test, serum IgG-EIA methods). All of the patients (100%) were found positive by WB-IgG, and 9 (39%) were positive by WB-IgA tests. H.pylori IgG and IgA antibodies showed reactivity with the different combinations of p120 (CagA), p95 (VacA), p29 (UreA) and p66 (UreB) antigens. The number of positive samples for CagA-IgG, VacA-IgG, UreA-IgG, CagA-IgA and UreA-IgA were 19, 17, 21, 8 and 4, respectively, while there were no positive sample for VacA-IgA. The antigenic distribution did not show any difference between age and sex of the patients. In our study the sensitivities of WB-IgG and WB-IgA methods were estimated as 100% and 39%, respectively, which were in accordance with the studies in literature. As a result, WB-IgG method appears to be useful and reliable for the detection of antibody profiles to H. pylori antigens and virulence factors, rather than routine and screening purposes, however the diagnostic value of WB-IgA method is low. It was thought that the results of our preliminary study should be confirmed by other studies including larger patient populations and control groups, in our country. PMID- 12652868 TI - [Evaluation of the methods used for the serologic diagnosis of brucellosis]. AB - Serological tests are the most widely used methods for the diagnosis of Brucella infections, since bacterial isolation techniques are time consuming. The conventional serological methods are the tube and slide agglutination tests, however enzyme immunoassays (ELISA) have gained importance recently as rapid diagnostic techniques. The aim of this study was to evaluate the performances of standard tube agglutination (STA, Wright test), slide agglutination (Rose-Bengal, RB) and ELISA-IgG and -IgM methods, in the serological diagnosis of brucellosis. For this purpose, Brucella antibodies have been searched in the serum samples of 184 patients (92 acute and 92 chronic brucellosis cases, of these 21 had positive blood cultures) and 20 healthy blood donors, who inhabit in the same region, as the control group. The positivity rates of STA (titers > or = 1/160), ELISA-IgG and -IgM in the patient group were found 83.7%, 61.9% and 49.5% respectively, while RB positivities for Brucella abortus and B. melitensis were 61.9% and 67.9%, respectively. In the control group STA and ELISA tests showed negative results, but RB for B. abortus and B. melitensis were positive in 25% and 30% of controls, respectively. Since ELISA and RB were found less sensitive than STA, it was concluded that STA is still the most reliable method for the serological diagnosis of brucellosis. PMID- 12652869 TI - [Polymerase chain reaction for Chlamydia trachomatis in urine specimens from patients with mucopurulent genital discharge]. AB - The aim of this study was to investigate the DNA of Chlamydia trachomatis by polymerase chain reaction (PCR) in the first-void urine samples of patients with mucopurulent genital discharge and to compare the results with the urethral/endocervical swab culture method. First-void urine samples from 56 patients (46 female, 14 male) and urethral swab samples from 14 male patients were tested by PCR. Additionally, shell-vial culture method was performed for the urethral/endocervical swab samples which were collected from 46 female and 14 male patients. Four (2 females, 2 males) of the patients (7.1%) showed positive results by both of the methods. In five (8.9%) of the urine samples, internal control tests were found negative, indicating the presence of amplification inhibitors, and the culture results of these patients were also negative. Since the PCR method (Cobas Amplicor CT, Roche Diagnostic Systems, NJ, USA) which was used in the study included internal control programme to identify inhibitors in urine, the sensitivity was improved. As a result, the perfect (100%) correlation between culture and PCR methods, lead to the conclusion that PCR is a rapid and reliable method for the detection of C. trachomatis in urine samples, however more detailed studies are necessary related to the sensitivity and specificity of PCR method. PMID- 12652870 TI - [Detection of cytomegalovirus DNA in immunosuppressed patients using qualitative and quantitative molecular methods]. AB - In this study, blood samples collected from 101 immunosuppressive patients were investigated for the presence of cytomegalovirus (CMV) DNA, with qualitative nested-polymerase chain reaction (PCR), and leukocytes obtained from these samples with quantitative hybrid capture assay (HCA). CMV-DNA was found positive in 32 (31.7%) and negative in 45 (44.5%) patients with both of the methods, and the agreement between the methods were estimated as 76.2%. The number of samples, which were PCR positive and HCA negative, were 24 (23.7%), while there were no samples which were PCR negative and HCA positive. All of the 56 CMV-DNA positive patients detected by PCR, were found positive for CMV-IgG, and 7 of them were also CMV-IgM positive. As a result, it was concluded that PCR is a practical and reliable method especially for the routine procedures for the investigation of CMV-DNA, however in cases which necessitate the detection of viral load, hybridization may be the preferable method. PMID- 12652871 TI - [Cloning and expression of the hepatitis B virus HBcAg gene in eukaryotic cells]. AB - In the present study, cloning of hepatitis B core antigen (HBcAg) gene and expression of the gene in eukaryotic cells have been reported. For this purpose, initially, HBcAg gene which was previously cloned into pUC19 plasmid, was excised by EcoRI ve HindIII from this plasmid and inserted into an eukaryotic expression vector pcDNA3. Afterwards, the resulting recombinant plasmid (pcDNA-HBc) was transfected into Vero cells, and the stable transfected cells (Vero-HBc) were selected in geneticin containing culture medium. The presence of HBcAg gene in Vero-HBc cells were confirmed by polymerase chain reaction and, the expression of HBcAg gene by Vero-HBc cells were tested in Western Immunoblotting assay. As a result, a 21 kDa protein reacting with anti-HBc antibodies which Vero-HBc cells indeed expressed, were detected at the end of this assay. PMID- 12652872 TI - [Standardization of neutralization tests using the COBL cell line and comparison with the particle agglutination test for measles serology]. AB - The aim of the present study was the detection and comparison of measles antibody titers with particle agglutination (PA) and neutralization (Nt) methods, in the sera samples of 364 subjects from different age groups. PA method was performed with a commercial test kit (Serodiameasles, Fujirebio Com. Japan), and Nt test which was standardized in this study, by using COBL (cord blood) cell lines, has been started to use in our laboratory as a reference method. As a result, antibody titers detected by PA were in parallel to the titers which detected by Nt test, and it was concluded that the differences in antibody titers would arise from the differences of test principles and viral antigens. PMID- 12652874 TI - [The rate of parasite detection over a 5-year period in the parasitology faculty laboratory at the Hacettepe University]. AB - The aim of this retrospective study was to investigate the rate of parasite positivity, in the samples which were sent to Parasitology Laboratory of Hacettepe University Medical School, between 1997-2001. A total of 58.150 specimens collected from subjects of whom 42% were adult and 58% were children, have been evaluated for the presence of parasites. Most of the samples (98%) were feces, and the remaining were sputum, cellotapes, blood and cyst materials. One or more parasites were detected in 2.117 (3.6%) of the specimens, and Giardia intestinalis (69.5%), Enterobius vermicularis (9.7%) and Taenia saginata (6.8%) were the most frequently encountered parasites. PMID- 12652873 TI - [The existence of contamination in enzymes used for the isolation of Aspergillus and Candida species DNA]. AB - Molecular biological methods which are widely used in different fields, have been replaced with conventional diagnostic tests for the early diagnosis of invasive fungal infections, recently. Polymerase chain reaction (PCR) is one of these methods, with high specificity and sensitivity, which is accepted throughout the world. However, the enzymes that are used for the isolation of target DNA, may be contaminated with the gene sequences of some other fungal species in the preparation steps and may affect the PCR results. The studies showed that, the contamination of these enzymes with any type of fungi during their production steps, leads false positive results in PCR tests. So, the additional studies are recommended for minimizing the contamination. The aim of this study was to search whether the enzymes necessary for the isolation of fungal target DNA, used in PCR, are contaminated or not. For this purpose, five different enzymes namely, Zymolase 20T (from Arthrobacter luteus, two examples from different companies), lyticase (from Arthrobacter luteus), lysing enzyme (from Trichoderma harzianum) and proteinase K (from Tritrachium album) have been investigated for the presence of contamination. As a result, both of zymolase 20T enzymes were found to be contaminated from some fungal species with the demonstration of 18S rRNA gene sequences, while the other enzymes were found non-contaminated. By the help of this method, the most suitable enzyme for PCR was chosen and fungal contamination was prevented. PMID- 12652875 TI - [Case report: a brain hydatid cyst]. AB - Hydatid cyst is a zoonotic infection caused by larval stage of Echinococcosis granulosus. Although any organ may be involved, bone, brain and heart involvements are uncommon. The brain involvement rate varies from 1% to 3% in hydatid disease. In this report, a 42 years old female patient has been presented in whom brain hydatic cyst was diagnosed by radiological and histopathological findings. PMID- 12652876 TI - [Case report: psoas abscess due to brucellosis]. AB - Skeletal system involvement is relatively a common complication of human brucellosis. However, muscular involvement is less frequent, and particularly psoas abscess is always secondary to spondylitis. In this case report, psoas abscess formation in a 70 years old female brucellosis patient complicated with spondylitis on vertebral corpus, has been presented. PMID- 12652877 TI - Light and shadow: reflections on environmental work. PMID- 12652878 TI - The dark side of consciousness and the therapeutic relationship. PMID- 12652879 TI - The dark side of consciousness. PMID- 12652880 TI - Reiki: a starting point for integrative medicine. PMID- 12652881 TI - Preliminary report on the use of Reiki HIV-related pain and anxiety. PMID- 12652882 TI - The effect of mirthful laughter on stress and natural killer cell activity. AB - CONTEXT: A recent survey of rural Midwestern cancer patients revealed that humor was one of the most frequently used complementary therapies. Psychoneuroimmunology research suggests that, in addition to its established psychological benefits, humor may have physiological effects on immune functioning. OBJECTIVE: To determine the effect of laughter on self-reported stress and natural killer cell activity. DESIGN: Randomized, pre-post test with comparison group. SETTING: Indiana State University Sycamore Nursing Center, which is a nurse-managed community health clinic in a mid-sized, Midwestern city. PARTICIPANTS: 33 healthy adult women. INTERVENTION: Experimental subjects viewed a humorous video while subjects in the distraction control group viewed a tourism video. MAIN OUTCOME MEASURES: Self-reported stress and arousal (Stress Arousal Check List), mirthful laughter (Humor Response Scale), and immune function (chromium release natural killer [NK] cell cytotoxicity assay). RESULTS: Stress decreased for subjects in the humor group, compared with those in the distraction group (U32 = 215.5; P = .004). Amount of mirthful laughter correlated with postintervention stress measures for persons in the humor group (r16 = -.655; P = .004). Subjects who scored greater than 25 on the humor response scale had increased immune function postintervention (t16 = 2.52 P = .037) and compared with the remaining participants (t32 = 32.1; P = .04). Humor response scale scores correlated with changes in NK cell activity (r16 = .744; P = 001). CONCLUSION: Laughter may reduce stress and improve NK cell activity. As low NK cell activity is linked to decreased disease resistance and increased morbidity in persons with cancer and HIV disease, laughter may be a useful cognitive behavioral intervention. PMID- 12652883 TI - The search for a biosensor as a witness of a human laying on of hands ritual. AB - CONTEXT: Intentional healing by laying on of hands is a popular complementary therapy. Previous studies of this therapy have been focused on the influence of laying on of hands with focused intention on the patient or on a biological model that took the place of the patient. OBJECTIVE: Exploring the line of thinking that the consciousness-mediated act of healing during a healer-patient ritual changes a consciousness field that could be detected in another living non-human organism that was present only as a witness and was not the object of any directed intention. DESIGN: A comparison of a biosensor's behavior during healer patient ritual treatments that were alternated by non-healing periods. SETTING AND PARTICIPANTS: An automatic device for measurement of ultra-weak emission of photons from algae was placed at the location of a healer during a series of experiments consisting of 36 healing sessions with human patients. Neither healer nor patients were aware of the type of measurements that took place. MAIN OUTCOME MEASURES: The number and periodicity of photon counts. RESULTS: Primary data analysis showed that the photon count distributions show some remarkable alterations during the ritual of healer-patient sessions. The data further suggest that during healing a shift in cyclical components of photon emission occurs. CONCLUSIONS: The significance of the experiment lies in the possibility to enter the discussion on a quantitative basis with respect to the relevance of the patient-healer relationship in intentional healing. PMID- 12652884 TI - Science and spiritual healing: a critical review of spiritual healing, "energy" medicine, and intentionality. PMID- 12652885 TI - Reiki--review of a biofield therapy history, theory, practice, and research. AB - Reiki is a vibrational, or subtle energy, therapy most commonly facilitated by light touch, which is believed to balance the biofield and strengthen the body's ability to heal itself. Although systematic study of efficacy is scant thus far, Reiki is increasingly used as an adjunct to conventional medical care, both in and out of hospital settings. This article will describe the practice and review the history and theory of Reiki, giving readers a context for the growing popularity of this healing modality. Programs that incorporate Reiki into the clinical setting will be discussed, as well as important considerations in setting up such a program. Finally, the research literature to date on Reiki will be reviewed and evaluated, and directions for future Reiki research will be suggested. PMID- 12652886 TI - An investigation into the efficacy of Scutellaria lateriflora in healthy volunteers. AB - Scutellaria lateriflora is an herbal medicine with long-standing traditional use as a relaxing nervine. There has been controversy in the literature with regards to its efficacy, and this study was designed to clarify its effectiveness in reducing anxiety, one of the phytotherapeutic indications. A double blind, placebo-controlled study of healthy subjects demonstrated noteworthy anxiolytic effects. The use of phytomedicines for the treatment of anxiety is reviewed, as is the published literature on S. lateriflora and its putative toxicity. PMID- 12652888 TI - Progress notes: University of Washington School of Nursing. PMID- 12652887 TI - Michael Lerner, PhD. Medicine and the environment. Interview by Bonnie Horrigan. PMID- 12652889 TI - The power of sound: ethnomedical tradition and modern science. PMID- 12652890 TI - Environmental ethics: finding a moral compass for human-plant interaction. PMID- 12652891 TI - Pediatric oncology patients find help and hope in New York City. PMID- 12652893 TI - Hemispheric modulatory influences on skin resistance response latency: unilateral stimulation, bilateral recording. AB - In order to evaluate the bilateral asymmetries in evoked electrodermal activity (EDA) and the use of EDA parameters as indices of relative hemispheric activation, skin resistance level (SRL), skin resistance response (SRR), and the SRR latency (L) were examined in 25 right-handed male subjects. We used bilateral recording with a unilateral stimulation (auditory click/patellar tendon tap) to assess the asymmetries of EDA variables related to the side of stimulation and to handedness. Although no significant asymmetry in C = SRRmax/SRL ratio was found, significant differences in latency were observed. Ipsilateral responses were 100 ms faster than contralateral responses for auditory stimulus, a result that can be explained in terms of contralateral delay of neuronal communication on EDA-l pathway. Response latency to reflex-motor activation was 60-100 ms shorter in the dominant hand, regardless of the stimulation site used. This result indicates that the reaction time of the EDA-2 pathway of the left hemisphere was shorter than the right for right-handed subjects. It is concluded that there is hemispheric asymmetry on EDA-2 pathway and that this asymmetry appears to be dependent on the preferred dexterity. These results have implications for the influence of both central and peripheral factors on EDA laterality and principally on laterality of response amplitude. Further, there is no single cortical mechanism modulating the asymmetries in the latency of this response. PMID- 12652892 TI - Enhancing the treatment of HIV/AIDS with Reiki training and treatment. PMID- 12652894 TI - Hypothalamic digoxin, cerebral dominance, and mitochondrial function/free radical metabolism. AB - The present study assessed the biochemical differences of free radical metabolism and mitochondrial function between right hemispheric dominant and left hemispheric dominant individuals. The following parameters were measured: (1) plasma HMG CoA reductase activity, (2) isoprenoid metabolites--digoxin and ubiquinone, (3) plasma magnesium and RBC membrane Na(+)-K+ ATPase activity; (4) lipid peroxidation products--malondialdehyde, hydroperoxides and conjugated dienes, and NO, (5) reduced glutathione, and (6) activity of superoxide dismutase, catalase, GSH peroxidase, and GSH reductase. The results showed that right hemispheric dominant individuals had (i) increased plasma HMG CoA reductase activity and elevated digoxin levels, (ii) decreased plasma magnesium and RBC membrane Na(+)-K+ ATPase activity, (iii) reduced ubiquinone levels, (iv) with increased levels of lipid peroxidation products and NO, (v) decreased levels of reduced glutathione and free radical scavenging enzymes, and (vi) increased tryptophan and reduced tyrosine levels. Left hemispheric dominant individuals had the opposite patterns. Right hemispheric dominance represents a hyperdigoxinemic state with membrane sodium-potassium ATPase inhibition and increased lipid peroxidation. Left hemispheric dominance represents the reverse pattern with hypodigoxinemic/membrane sodium-potassium ATPase stimulation and decreased lipid peroxidation. Cerebral dominance can regulate mitochondrial function and free radical metabolism. PMID- 12652895 TI - Hypothalamic digoxin, hemispheric chemical dominance, and endocrine/metabolic/cellular regulation. AB - The hypothalamus produces an endogenous membrane Na(+)-K+ ATPase inhibitor and regulator of neurotransmission, digoxin. Digoxin, a steroidal glycoside, is synthesized by the isoprenoid pathway. In view of the reports of elevated digoxin levels in metabolic syndrome X with high body mass index, the isoprenoid-mediated pathway biochemical cascade was assessed in individuals with high and low body mass index. It was also assessed in individuals with differing hemispheric dominance to find out the relationship among digoxin status, body mass index, and hemispheric dominance. The isoprenoid pathway metabolites, tryptophan/tyrosine catabolic patterns, glycoconjugate, and free radical metabolism, as well as membrane composition, were assessed. In individuals with high body mass index, an upregulated isoprenoid pathway with increased digoxin levels, increased glycoconjugates, and dolichol levels, reduced lysosomal stability, low ubiquinone levels with increased free radical generation, and increased membrane cholesterol:phospholipid ratio were observed. The reverse patterns were seen in individuals with a low body mass index. The patterns in individuals with a high body mass index and low body mass index correlated with right hemispheric dominance and left hemispheric dominance, respectively. Hemispheric dominance and digoxin status regulated the differential metabolic pattern observed in individuals with high and low body mass index. Hypothalamic digoxin/cerebral dominance can regulate the metabolic/endocrine function, as well as the structure/function of cellular organalle. PMID- 12652896 TI - Hypothalamic digoxin, cerebral dominance, and membrane biochemistry. AB - The present study assessed the biochemical differences in membrane composition/function between right hemispheric dominant and left hemispheric dominant individuals. The HMG CoA reductase activity, serum isoprenoid metabolite -digoxin--serum magnesium, and the RBC membrane composition/Na(+)-K+ ATPase activity were studied. The results showed that right hemispheric dominant individuals had (i) increased HMG CoA reductase activity and elevated digoxin levels, (ii) decreased RBC membrane Na(+)-K+ ATPase activity and serum magnesium levels, and (iii) increased cholesterol:phospholipid ratio of RBC membranes with reduced membrane glycoconjugates. Left hemispheric dominant individuals had the opposite patterns. Right hemispheric dominance represents a hyperdigoxinemic state with membrane sodium-potassium ATPase inhibition. Left hemispheric dominance represents the reverse pattern with hypodigoxinemia and membrane sodium potassium ATPase stimulation. Cerebral dominance can regulate membrane structure/function. PMID- 12652897 TI - Hypothalamic digoxin, cerebral dominance, and Golgi body/lysosomal function. AB - The present study assessed the biochemical differences of glycoconjugate metabolism between right hemispheric dominant and left hemispheric dominant individuals. The isoprenoid metabolites--digoxin and dolichol, glycoconjugates, and lysosomal enzymes--were studied. The results showed that right hemispheric dominant individuals had increased (i) HMG CoA reductase activity and elevated digoxin levels, and (ii) increased dolichol and glycoconjugate levels with reduced lysosomal stability. Left hemispheric dominant individuals had the opposite patterns. Right hemispheric dominance represents a hyperdigoxinemic state with membrane sodium-potassium ATPase inhibition. Left hemispheric dominance represents the reverse pattern with hypodigoxinemia and membrane sodium potassium ATPase stimulation. Cerebral dominance can regulate glycoconjugate metabolism (golgi body/lysosomal function). PMID- 12652898 TI - The relationship between magnitude of cerebral activation and intensity of emotional arousal. AB - There is considerable knowledge concerning the cerebral localization of positive and negative emotions; however, relatively little is known about the cerebral representation of subjective emotional intensity. Indeed, few studies to date seem to have been conducted on the cerebral effects of increased emotional intensity. The purpose of the present investigation was to investigate the relationship between the intensity of cerebral activation and emotional arousal using quantitative electroencephalography (QEEG). Participants were asked to recollect an angry memory while QEEG was recorded from 19 electrode sites arranged according to the International 10/20 System. Significant positive correlations were found between subjective intensity of angry memories and changes in low beta (13-21 Hz) and high beta (21-32 Hz) magnitude (microV) at the F8, T6, and O1 electrode sites among the male participants. Additionally, a significant positive correlation between emotion intensity and changes in high beta at the T6 electrode site was found among female participants. Results are discussed in terms of implications for emotional regulation and dysfunction. PMID- 12652899 TI - Index, summary, and subtest discrepancy scores on the WAIS-III in postacute traumatic brain injury patients. AB - WAIS-III profile interpretation typically involves the analysis of disparities between summary and subtest scores. Determination of clinically meaningful differences has been guided by published rates of discrepancies based upon the normative sample. However, noting that a particular profile discrepancy is uncommon among normals only represents one step in the interpretive process. Rates of such pairwise differences have yet to be published for clinical samples. In order to begin filling this void in the literature, we present frequencies of composite and subtest differences from a postacute traumatic brain injury sample. The practical utility of these tables, in conjunction with the normative tables, is demonstrated. PMID- 12652900 TI - Trail making test and malingering among substance abusers. AB - The Trail Making Test (TMT) is often used for screening cognitive impairments in substance abusers. A possible limitation of the TMT in clinical settings is that substance abusers may malinger and give poor effort. In this study, previously validated cutting scores for malingering were applied to a sample of 7689 substance abusers (primary drug of abuse-number of subjects: Alcohol-1000, Marijuana-259, Hallucinogen-128, Cocaine/crack-4306, Heroin-1548, Narcotics/other opiates-191, Sedatives-72, Amphetamines-185) in drug abuse treatment programs. A mixed race sample was drawn from electronic files of data from the Drug Abuse Treatment Outcome Study (DATOS). The DATOS was a naturalistic, prospective cohort study that collected data from 1991-1993 in 96 programs in 11 cites in the United States. Data were analyzed to determine the number of substance abusers that fell beyond the preset malingering cutting scores on the TMT in this very large sample of substance abusing patients in treatment settings. The TMT variables of seconds to complete Part A and Part B, and the ratio score of Part B divided by Part A (B/A), ranged from no subjects beyond the preset cutting score for Part B to 2.28% (175 of 7689 subjects) for Part A to 9.74% (749 of 7689 subjects) for the ratio score. Most substance abusers fell within preset cutting score ranges, a finding that suggests that their scores are valid. Another interpretation of the data, however, is that the cutoff scores were not particularly sensitive to biased responding. Further research is indicated. PMID- 12652901 TI - ERPs associated with familiarity and degree of familiarity during face recognition. AB - Event-related potentials (ERPs) triggered by three different faces (unfamiliar, famous, and the subject's own) were analyzed during passive viewing. A familiarity effect was defined as a significant difference between the two familiar faces as opposed to the unfamiliar face. A degree of familiarity effect was defined as a significant difference between all three conditions. The results show a familiarity effect 170 ms after stimulus onset (N170), with larger amplitudes seen for both familiar faces. Conversely, a degree of familiarity effect arose approximately 250 ms after stimulus onset (P2) in the form of progressively smaller amplitudes as a function of familiarity (subject's face < famous face < unfamiliar). These results demonstrate that the structural encoding of faces, as reflected by N170 activities, can be modulated by familiarity and that facial representations acquire specific properties as a result of experience. Moreover, these results confirm the hypothesis that N170 is sensitive to face versus, object discriminations and to the discrimination among faces. PMID- 12652902 TI - The major component of the cellulosomes of anaerobic fungi from the genus Piromyces is a family 48 glycoside hydrolase. AB - Sequencing of two cDNAs from the anaerobic fungi Piromyces equi and Piromyces sp. strain E2 revealed that they both encode a glycoside hydrolase (GH) family 48 cellulase, containing two C-terminal fungal dockerin domains. N-terminal sequencing of the major component of the Piromyces multi-enzyme cellulase/hemicellulase complex, termed the cellulosome, showed that these 80 kDa proteins corresponded to the GH family 48 enzyme. These data show for the first time that GH family 48 cellulases are not confined to bacteria, and that bacterial and fungal cellulosomes share the same pivotal component. PMID- 12652903 TI - Cloning and characterization of a Helicobacter bizzozeronii urease gene cluster. AB - The urease gene cluster from Helicobacter bizzozeronii was cloned and sequenced. A genomic library was constructed in a lambda-ZAPII vector using TSP5091-digested H. bizzozeronii chromosomal DNA. Four overlapping recombinant bacteriophages carrying the H. bizzozeronii urease genes were identified by using a fragment of H. bizzozeronii ureB as a probe. Sequence analysis of two clones (pHB1 and pHB3) revealed seven open reading frames encoding proteins with predicted masses of 26.5, 60.3, 21.7, 19.5, 28.6, 21.7 and 29.6 kDa representing the structural genes, Urease A and B and its accessory genes, urease I, E, F, G and H, respectively. In addition, three open reading frames upstream of the ureA gene encoding a putative tRNA transferase, a putative Glucose inhibited division protein B (GidB) and a protein with unknown function were also identified. A clone (pHB5) containing a complete urease gene cluster was constructed. The homologue analysis revealed that UreA polypeptide exhibited 64-90% identity to that of Helicobacter heilmanii, Helicobacter felis, Helicobacter pylori, Helicobacter mustelae and Helicobacter hepaticus. UreB polypeptides exhibited 76.8-96% identity to that of H. heilmanii, H. felis, H. pylori, H. mustelae and H. hepaticus. The UreI, E, F, G and H also showed 44-86% identity to that of H. pylori. Among these accessory genes, UreE had a lowest percentage identity to that of H. pylori. PMID- 12652904 TI - Cloning and characterization of an iron regulated locus, iroA, in Salmonella enterica serovar Choleraesuis. AB - To identify genes belonging to the Ferric update regulator (Fur) regulon of Salmonella enterica serovar Choleraesuis, the Fur titration assay (FURTA) was used to screen a genomic library for Fur promoters and iron-regulated genes. Fifteen FURTA positive clones were identified from this assay. DNA sequence analysis of these clones showed that 11 out of 15 clones had a Fur binding site (Fur box), and 6 of these clones showed homology to the iron-regulated genes of S. enterica serovar Typhi and/or E. coli. One of these clones (pSC4) was homologous to the iroB gene of the iroA locus of S. enterica serovar Typhi. The iroA locus of S. enterica serovar Choleraesuis was cloned from a lambda-dash library and subjected to DNA sequencing. The complete nucleotide sequence of 9848 bp of the iroA locus of S. enterica serovar Choleraesuis consists of iroB, C, D, E and N genes, which are transcriptionally regulated by Fur. The amino acid sequence of IroB, C, D, E and N was 95%, 86, 89, 96 and 96% identity to that of S. enterica serovar Typhi. The IroN gene was homologous to the family of TonB dependent outer membrane receptors and the putative virulence factor, IroNE. coli, of the extraintestinal pathogenic E. coli. The convalescent porcine sera contained antibodies against the three major iron-regulated outer membrane proteins of S. enterica serovar Choleraesuis. An insertional inactivation of the iroN gene of S. enterica serovar Choleraesuis by allelic exchange resulted in the loss of expression of the 78 kDa protein. However, this mutant had a similar LD50 to mice compared to the parent strain when given intraperitoneally. PMID- 12652905 TI - Analysis of rILERS, an isoleucyl-tRNA synthetase gene associated with mupirocin production by Pseudomonas fluorescens NCIMB 10586. AB - Some strains of Pseudomonas fluorescens produce the antibiotic mupirocin, which functions as a competitive inhibitor of isoleucyl-tRNA synthetase (ILERS). Mupirocin-producing strains of P. fluorescens must overcome the inhibitory effects of the antibiotic to avoid self-suicide. However, it is not clear how P. fluorescens protects itself from the toxic effects of mupirocin. In this report, we describe a second gene encoding isoleucyl-tRNA synthetase (rILERS) in P. fluorescens that is associated with the mupirocin biosynthetic gene cluster. Random mutagenesis of the mupirocin-producing strain, P. fluorescens 10586, resulted in a mupirocin-defective mutant disrupted in a region with similarity to ILERS, the target site for mupirocin. The ILERS gene described in the present study was sequenced and shown to be encoded by a 3093 bp ORF, which is 264 bp larger than the ILERS gene previously identified in P. fluorescens 10586. rILERS from P. fluorescens is most closely related to prokaryotic or eukaryotic sources of ILERS that are resistant to mupirocin. Interestingly, the relatedness between rILERS and the ILERS previously described in P. fluorescens 10586 was low (24% similarity), which indicates that P. fluorescens contains two isoforms of isoleucyl-tRNA synthetase. PMID- 12652907 TI - Complete cDNA coding sequence of the HLA-DRB1*1405 allele. AB - Cloning report of complete coding region of HLA-DRB1*1405 has never appeared in the literature. We have isolated and sequenced it from a B cell line with a DRB1*1405/1502 genotype. The sequence information of this gene and previously published DRB1 genes for the other DR serotypes facilitated us to find that the cytoplasmic domain sequences of DRB1 genes fall into three groups. It is known that antigen presenting cells are differentially activated when products of different MHC class II genes are triggered. Functional analyses would discern if these allelic differences in the cytoplasmic domain are of functional significance. PMID- 12652906 TI - Identification of a syndecan 4 pseudogene. AB - The syndecan family of heparan sulfate proteoglycans participates in cellular activation through interactions with growth factors, extracellular matrix, and other molecules. The family consists of four proteins that share sequence homology within their cytosolic domains. Here we report that a 5.8 kb region of human chromosome 22q12.2 contains multiple segments that share greater than 80% sequence homology to the syndecan 4 transcript, including homology to 443 nucleotides of the syndecan 4 coding region. Three pieces of evidence indicate that the chromosome 22 sequences are a syndecan 4 pseudogene. First, single nucleotide gaps need to be inserted into the chromosome 22 sequence in order to maintain maximal alignment to the syndecan 4 coding sequence, and this introduces stop codons into a deduced amino acid sequence. Second, the total length of chromosome 22 containing the homologous sequences is compressed when compared to the genomic organization of the complementary syndecan 4 sequences. Third, the 5.8 kb chromosome 22 sequence contains multiple Alu and other repetitive sequences, and this is a property of pseudogenes. Both RT-PCR and RNase protection assays indicated that the syndecan 4 pseudogene is transcribed in human umbilical vein endothelial cells. PMID- 12652908 TI - Isolation and sequence analysis of the rat dihydrolipoamide succinyltransferase gene. AB - The dihydrolipoamide succinyltransferase (DLST) gene of the alpha-ketoglutarate dehydrogenase complex (alpha-KGDC) was isolated from a rat genomic DNA library and sequenced. This gene was composed of 15 exons and 14 introns like the human DLST gene. Sequence analysis of the promoter-regulatory region of the rat DLST gene-(Dlst) showed the possible presence of a CAAT box-sequence and of the sequences for an AP-2 site and three Sp1 sites, but no TATA box-sequence was evidenced. The nucleotide sequences of introns 1 and 4 of the rat Dlst were significantly homologous to those of introns 1 and 4 of the human DLST gene. The sequence analysis of the rat Dlst suggested that the exon coding for the E3- and/or E1-binding domain may have been lost from the gene during evolution in eukaryotic DLST, possibly after mitochondrial symbiosis because prokaryotic DLST possesses the E3- and/or E1-binding domain. PMID- 12652909 TI - cDNA and deduced amino acid sequences of ferret preproendothelin-2 and phylogenetic analysis. AB - Ferret preproendothelin-2 (PPET2) cDNA was cloned from intestinal tissue by reverse transcription-polymerase chain reaction (RT-PCR) in conjunction with 5'- and 3'-rapid amplification of cDNA ends (RACE). The cDNA comprises 1230 bp, excluding the poly(A) tail, and has 534 bp of open reading frame encoding a putative polypeptide of 178 residues, in which a 21-amino acid mature endothelin 2 (ET2) peptide as well as a 24-amino acid putative signal peptide and a 16-amino acid ET2-like peptide were found. The homology of the full-length cDNA sequence of ferret with those of horse, human, mouse, or rat was 75.6, 71.6, 65.4 or 65.1%, respectively, and the homology of the coding region was 85.1, 81.6, 78.1 or 75.3%, respectively. Phylogenetic analysis among ferret, horse, human, mouse, rat and dog showed that ferret has a closer relationship to dog than to the other mammals. PMID- 12652910 TI - Cloning and characterization of a gene encoding glutathione-regulated potassium efflux system protein KefKL from the endosymbiont Wolbachia. AB - The maternally inherited intracellular symbiont Wolbachia is well known for inducing a variety of reproductive and developmental abnormalities in the diverse arthropod hosts it infects. It has been implicated in causing cytoplasmic incompatibility (CI), parthenogenesis, feminization of genetic males and male killing in different hosts. However, the molecular mechanisms by which this fastidious bacterium causes these abnormalities have not yet been determined. In our study, representational difference analysis (RDA) was used to analyze the genomic difference between different Wolbachia strains. A gene encoding glutathione-regulated potassium-efflux system protein KefKL from Wolbachia in Drosophila simulans Riverside (w Ri) was isolated. The homologous genes from Wolbachia in Drosophila melanogaster yw67c23 (wMel) and Wolbachia in Drosophila melanogaster CantonS (wMelCS) were also cloned and sequenced. Sequence analysis showed that these deduced amino acid sequences contained two important motifs: Na+/H+ antiportor and NAD binding domain, which shared conserved sequences among different strains. Considering the crucial function of KefKL for ionic homeostasis, this gene might play an important role in Wolbachia physiology. Further study indicated that there was no homologue detected from Wolbachia in Drosophila simulans DSW/Mau (wMa) and Wolbachia in Drosophila simulans Noumea (wNo). Whether Wolbachia contained KefKL (or the homologous gene) was consistent with the phylogenetic studies using wsp sequences, which showed that wMa and wNo were grouped into one branch, while w Ri, wMel and wMelCS were more closely related. PMID- 12652911 TI - Primary structure of the histone 2B gene in the white root rot fungus, Rosellinia necatrix. AB - The nucleotide sequence of the histone 2B (H2B) gene in the white root rot fungus, Rosellinia necatrix, was determined. The gene has two introns in the coding region at positions conserved in the Neurospora crassa and Aspergillus nidulans H2B genes, but the third intron present in the H2B gene from N. crassa and A. nidulans is absent in the R. necatrix H2B gene. The amino acid sequence of the coding region of the R. necatrix gene resembled that of N. crassa and A. nidulans. Therefore, the third intron in the H2B gene of N. crassa and A. nidulans may have been inserted into the present position after species diversification. PMID- 12652912 TI - Cloning and sequencing of the Thermus aquaticus glycerol facilitator gene. AB - The gene glpK, encoding glycerol kinase of Thermus aquaticus has been identified [Biosci. Biotechnol. Biochem., 62 (1998) 2375-2381]. In the present work, the nucleotide sequence of glpFK operon and the gene glpF encoding glycerol facilitator were determined. T. aquaticus GlpF was predicted to contain 272 amino acids with six putative transmembrane segments and two half-membrane-spanning segments that contained the motif Asn-Pro-Ala, respectively. The amino acid residues involved in the discrimination of glycerol were deduced to be Trp44, Tyr182, and Arg188. PMID- 12652913 TI - [Surgical complications following pancreaticoduodenectomy: results of a single center experience]. AB - Pancreaticoduodenectomy represents the only therapeutic option for cefalo pancreatic and periampullary cancers. Surgical and anaesthesiological techniques development over the last twenty years has granted an operative mortality decrease. However, surgical morbidity is still high, with an incidence of 30-50%. A 20 year experience of a single Centre is examined retrospectively: 121 patients underwent pancreatic resection with radical intent. Type of operation or re operation, operative mortality within 30 days, general and surgical morbidity, postoperative hospital stay were analysed. Average recovery time was 24 days (range 12-65); operative mortality was 5.8% (7/121); general morbidity, including medical and surgical complications, was observed in 47 patients (38.8%). Pancreatic fistula occurred in 16 patients (13.2%); ten of these underwent a second operation. Patients who underwent pancreaticoduodenctomy were divided as follows: 76 pts. received a pylours-preserving pancreaticoduodenectomy and 45 a Whipple's resection. Neither surgical complications incidence nor mortality rate were significantly different between the two groups. Postoperative complications following pancreaticoduodenectomy are still frequent and severe. In particular, pancreatic fistula represents the most relevant complication following pancreaticoduodenectomy. The Authors suggest that standard and meticulous surgical procedures together with continued efforts to improve postoperative follow-up, support early detection of complications and improvement of results in most patients. PMID- 12652914 TI - [Gastrointestinal stromal tumors: report of a clinical case and review of the literature]. AB - In this case report, the Authors describe a case of stromal gastric tumour, in a male 65 years old, who presented gastrointestinal bleeding. Gastro-Intestinal Stromal Tumors (GISTs) are neoplasm with an incidence of 1-3 per cent of the digestive tract malignant neoplasms. The rarity of this disease, its visceral wall localization, the histopathological characteristics make the diagnosis difficult. Moreover there is no correlation between the behaviour of these neoplasms and the histologic features. Surgery represents the main treatment for GISTs based on complete resection, followed by a long-term follow-up. Chemotherapy and radiotherapy don't seem to play a crucial role in the treatment of these neoplasms. A new treatment with inhibitors of the tyrosinekinase is under discussion. Follow-up represents the only way to evaluate the effective behaviour of the disease, due to the lack of classic prognostic factors impact. PMID- 12652915 TI - [Pica: a clinical case and therapeutic problems]. AB - Pica is a psychotic disorder characterized by compulsive ingestion of nonfood substances; the primary treatment is the prevention with antipsychotic therapy. When the patient ingests foreign body, endoscopical removal is recommended. Surgical treatment is necessary when an acute abdomen is present, with intestinal occlusion and/or bowel perforation. The Authors report a peculiar case of pica, and analyse the problems of organization and assistance, besides surgical approach, also on the base of international literature. PMID- 12652917 TI - [Cecal volvulus]. AB - A retrospective study on 18 patients with cecal volvulus surgically treated was made. Demographics and clinical data, as well as treatment were determined from clinical reports. The operative procedures employed were cecostomy (56%), cecopexy (22%) and right colectomy (22%). The length of follow up averaged 63 months and there was one recurrence. The Authors suggest that cecostomy should be employed in patients with viable bowel, and resection should be limited to cases with gangrene. PMID- 12652916 TI - [Retroperitoneal tumors: a case of liposarcoma]. AB - The Authors report a case of retroperitoneal sarcoma, in the histological subtype "liposarcoma", occasionally diagnosed in a 57 year old woman, who presented aspecific symptoms, characterized by dry cough, vomiting and chest pain. Chest TC showed a neoplasm of the retroperitoneum, close to liver, right colon, duodenum and right kidney. The arteriography of the right renal artery showed a neovascolarization inside the mass. A surgical resection was performed. The histological examination confirmed the diagnosis of liposarcoma. Liposarcomas account for 30% of the retroperitoneal soft tissue sarcomas; symptoms and diagnosis are late and the surgical resection of primary neoplasm, metastases and local recurrences representing the only therapeutic choice. PMID- 12652918 TI - [Recurrence after hernioplasty according to Lichtenstein: analysis of the cause]. AB - The performance of surgical interventions of tension-free hernia repair has certainly reduced the recurrence rate in comparison with the previous techniques; notwithstanding this it is not uncommon to observe some recurrences also after a Lichtenstein hernia repair. The aim of this study is that to analyse the causes. In the last 2 years, 42 patients (mean age of 68 years) with recurrent hernia have been operated by Lichtenstein technique; 11 of the 42 patients had been treated before with an useful mesh hernia repair. In 8 of these patients the recurrence was produced by an insufficient medial extension of the mesh; in the other 3 patients the cause was the presence of an unrecognized indirect hernia in patients operated for a direct inguinal hernia. All the patients treated have been submitted to a 18 months time of follow-up. PMID- 12652919 TI - [Atrial fibrillation and hyperthyroidism: results after thyroidectomy]. AB - Hyperthyroidism patients can develop atrial fibrillation (AF) with a frequency range between 10 and 25%. The Authors consider 256 hyperthyroid patients operated in their Department, from 1985 to 2001. Before and after surgery, thyroid examination, routine blood tests and common diagnostic evaluations were performed. The follow-up has been 12 months and during this period a L-Thyroxine (50-200 mg/die) therapy has been arranged. Before surgery 59 patients (23%) showed an AF associated with palpitations (76%), asthenia (66%), tremors (54%) and dyspnea (43%). The onset of AF was less than 1 months in 18% of cases, less than 3 months in 30% and chronic in remaining 52%. The 87% of patients was already pharmacologically treated. All the patients were treated with anticoagulants, with a INR value between 2 and 3. After the surgery, 47% of patients presented a restoration of the sinusal rhythm, while the 53% showed a stronger therapeutic responsiveness to specific antiarrhythmic therapy. Treatment of FA hyperthyroid patients with FA tends to normalize the function of the thyroid gland (euthyroidism). In our experience the restoring of sinusal rhythm usually develops in patients with 50 years of age (45 +/- 5 years), while in older patients AF tends to be resistant to treatment. PMID- 12652920 TI - Sentinel lymph node biopsy for sparing axillary dissection in selected patients with breast cancer. AB - Axillary lymph nodal status in breast cancer remains one of the more important prognostic factors. In early breast cancer axillary lymph node metastasis are found only in 10-18%. It can be deduced that in all these patients a complete axillary dissection is an overtreatment. The concept of sentinel lymph node (SN) was applied to breast cancer. Of course if SN examination gives negative findings, the patient will avoid axillary lymphadenectomy. 134 patients with localized breast cancer were evaluated for enrollment into the study. In 40 (29.8%) patients lymphoscintigraphy was performed together with an injection of vital dye to identify the SN, in 94 (70.1%) only vital dye was utilized. The mapping procedure was successful in 129 cases (96.2%). In our study there was concordance between SNs and axillary nodes in 120 out off 124 cases (96.7%). The false-negative rate was 4.8% (4/83). The overall sensitivity of the SN biopsy was 91.1% (41/45), with a negative predictive value of 95.1% (79/83). Five patients had SN negative and they decided do not undergo axillary lymphadenectomy. This study demonstrates that accurate SN identification was obtained combining lymphoscintigraphy and blue dye. Moreover, each method requires a suitable learning curve. After an accurate training, complete axillary lymphadenectomy can be avoided in selected patients. PMID- 12652921 TI - [Laparoscopic approach versus laparotomy for suspected acute appendicitis]. AB - Acute appendicitis is one of the most common surgical disease but, in spite of the progression diagnostic imaging, a definite diagnosis is frequently difficult and often is based in essentially clinical grounds. The Authors retrospectively analyze the results of conventional laparotomic appendectomy (CLA) and videolaparascopic appendectomy (VLA) as performed by two teams of their Department of Surgery. Between January 2000 and November 2001, 156 patients, age ranging from 3 to 67 yrs, underwent surgery because suspected acute appendicitis; 96 patients underwent VLA and 60 patients underwent CLA; a diagnosis of acute appendicitis was confirmed in 142 cases (91%). The patients who had a confirmed diagnosis of acute appendicitis were grouped according to clinical-pathologic criteria into follicular, gangrenous and complicated appendicitisi; operating time, complications and length of hospital stay were compared among the three groups. VLA resulted superior to CLA in terms of diagnostic performance: among the patients with suspected acute appendicitis, VLA allowed to establish a different diagnosis in 41% as compared to 13% with CLA. In two cases (2.08%) VLA was turned into CLA. There were non differences between VLA and CLA in terms of surgical operating time and hospital stay. A significantly higher incidence of surgical complications, such as wound infections and pelvic abscesses, was observed in the CLA group as compared to VLA. The Authors conclude that VLA allows a complete exploration of the abdomen, a better localization of the appendix and an easier diagnosis of unexpected conditions or pathologic processes associated with the clinical appendicitis; further, VLA is gravated by fewer complications. PMID- 12652923 TI - [Extracorporeal shock wave lithotripsy in the treatment of sialolithiasis. Long term results]. AB - The Authors report the results of a short term follow-up in 28 patients treated for sialolithiasis. Extracorporeal lithotripsy is safe, effective and minimally invasive therapeutical option. Open surgery is reserved to cases of failure of noninvasive procedure. PMID- 12652922 TI - [Periareolar subcutaneous quadrantectomy: a new approach in breast cancer surgery]. AB - Plastic and oncological breast surgery are becoming more and more closer as one surgical treatment. The term "oncoplastic surgery" refers to the use of plastic surgery techniques in breast cancer surgery, in order to avoid and to correct the adverse aesthetic findings. The care of cosmetic sequelae of breast cancer surgery has reached an important therapeutic role for psychological consequences of disease and because of the higher patients expectations of a good aesthetic result. Considering the concept of oncoplastic surgery, since 1999 the Authors began to use a periareloar approach in the breast conserving therapy (BCT), associated to axillary dissection performed through the same periareolar incision. This technique, original from the oncological point of view, is not different from the traditional quadrantectomy in the extension of the glandular resection, while the skin may be preserved in according to the conventional protocols of BCT. Oncological and aesthetic results have proved to be safe and satisfactory. PMID- 12652924 TI - [Health policy: with more individual responsibility out of the co-payment trap]. PMID- 12652926 TI - [Obstructive sleep apnea. Insurance legal regulation and the significance for expert assessment]. AB - Nocturnal breathing disorders are a common problem and affect about 500,000 adults in Germany. Obstructive sleep apnea is associated with an elevated cardiovascular risk, whereby arterial hypertension and coronary artery disease are frequent. Furthermore, in patients with sleep apnea excessive daytime sleepiness results in a high rate of accidents in traffic and work. Nocturnal continuous positive airway pressure is an effective treatment of obstructive sleep apnea. For social insurance agencies, immense direct and indirect costs arise from the sequel and accompanying illness of untreated sleep apnea. PMID- 12652925 TI - [Cancer fatigue syndrome]. AB - Tumour-related fatigue, which occurs in almost all tumour patients under therapy- very often even years after a successful therapy--has a considerable impact not only on the quality of life, but apparently also on the course and outcome of treatment. There is little known about where cancer fatigue syndrome comes from. Medical research is required particularly for the continuous states of tiredness persisting for years. The questionnaires currently in use permit an accurate evaluation of the existence and the course of tumour-related fatigue. The treatment of the cancer fatigue syndrome includes the correction of underlying metabolic disorders as well as--even more importantly--physical training and psycho-oncological measures in order to better cope with the disease. Many questions, however, are left open, for instance the impact of cancer fatigue syndrome on disability pensions of tumour patients. The German Fatigue Society (DfaG) was therefore founded in March 2002 in order to find out the correlations between these questions and the cancer fatigue syndrome. PMID- 12652927 TI - [Significance of environmental medicine from the viewpoint of the medical review board of the social health insurance program]. AB - Tasks concerning environmental medicine are a significant aspect of the expert work done by the medical review board of the social health insurance fund. Thus far there are no commonly accepted theories and/or criteria with regard to the cause of environmental incompatibilities, nor are there generally accepted criteria/standards for clinical diagnostic procedures and therapy. Problems arise from the fact that the field of environmental medicine not only offers scientifically accepted and verified diagnostic and therapeutic methods, but also numerous unconventional procedures without verified validity. The decision of the scientific expert has to be based on the legal principles of social legislation and jurisdiction. His/her opinion must be competent, objective and independent. Further research is urgently needed to improve the scientific data pool. With it, well-grounded methods and standards can be offered. PMID- 12652928 TI - [The so-called trauma criterium (A--Criterium in DSM-IV) of post-traumatic stress disorder and its significance for social and legal insurance (II)]. AB - Since the diagnosis post traumatic stress disorder (PTSD) was included in the American diagnosis system DSM-III in 1980, the ongoing scientific discussion has led to a shift in the definition of the so called A-criterion (trauma criterion) away from the general theory that the disease-causing event has to be outside normal human experience towards the point of view that it is more or less determined by the subjective experience of the individual. Without wishing to become involved in the discussion, in the publication the author tries to explain, on the basis of references to basic concepts found in insurance law (accident, initial damage, suitability of event, objectivity, epidemiological considerations), that in social insurance as well as in non-life insurance an objective insured event which has a severe emotional impact on the individual is a prerequisite for payment. The conclusion is that not every mental disorder which occurs after an insured event and which has quite rightly been classified as PTSD (or partial PTSD) on the basis of the symptoms involved in accordance with ICD-10/DSM-IV (criteria B-D) necessarily results in payment under insurance law. PMID- 12652929 TI - [Framework for qualified inpatient withdrawal treatment of alcoholics in North Rhine-Westphalia]. AB - The programme against addiction, set up by the Ministry for Women, Family, Youth and Health of North Rhine Westphalia, Germany and with broad participation from all groups concerned (MfjFG 1999), seeks the improvement of qualified in-patient withdrawal-treatment for alcoholics. Under the leadership of a district administration, a congress comprising seven one-day workshops was held for representatives of the Ministry, health insurance funds, pension insurance carriers and physicians. They agreed on the basic concept outlined below. Low threshold access, diversity of essential treatment goals, financial obligation on the part of the regular health insurances, nursing staff, physicians, social workers, and non-verbal therapists according to the decree for psychiatric hospitals in Germany (Bundesregierung 1990), a regular in-patient-period for uncomplicated detoxification and motivation of up to 21 days, prolonged in justified cases up to six weeks, and the integration in the regional help-system for persons with substance-use related problems are essential features of this concept. PMID- 12652930 TI - [Malnutrition in elderly patients of an acute care clinic]. AB - Malnutrition is associated with a higher complication rate, a higher morbidity and mortality. In-hospital patients consume up to 75% fewer calories than they would need, according to studies in international literature. For German general hospitals data are not yet available on this topic, especially for elderly people. Here we publish a prospective study of 166 persons, aged at least 70 years. According to international data in German general hospitals, elderly people consume only 50% of calories they need. There is evidence that malnutrition causes longer hospital stays, which not only adversely affects the patients themselves but also involves a considerable cost factor. PMID- 12652931 TI - [Intrinsic function of the state and change due to a global society]. PMID- 12652932 TI - [Study of the "Health Telephone" Medical Call Center]. PMID- 12652933 TI - [Comment on N.-A. Sittaro: Current status and future of insurance medicine]. PMID- 12652935 TI - [Comment on N.-A. Sittaro: Current status and future of insurance medicine]. PMID- 12652934 TI - [Comment on N.-A. Sittaro: Current status and future of insurance medicine] ]. PMID- 12652936 TI - [The opiate system in the regulation of central hemodynamics]. PMID- 12652937 TI - [On the mechanisms of radiobiological events in relation to the regulatory role of p53 gene and protein]. AB - The paper analyzes the topical problems of radiobiology in the light of the present-day data on the molecular biology and biochemistry of p53 protein that is an integrator of stress signals from various damaging exposures and that fulfills the function of genome guard by regulating the checking points of a cellular cycle, DNA reparation, and apoptosis. It also considers the mechanisms of radiation cell death and radiosensitivity/radioresistance in the light of data on p53 protein, as well as the problems of searching for antiradiation agents, the radiation-induced instability of genome, the biological aftereffects of small dose radiation, as well as radiation-induced carcinogenesis due to the important regulatory role of p53 protein. The lines of further studies of the above problems are outlined to refine the understanding of pathogenetic processes in radiation damages, to extend the therapeutic, diagnostic, predictive capacities of clinical radiobiology and radiation medicine. PMID- 12652938 TI - [The opiatergic link between the antiarrhythmic effect of adaptation and hypoxia in the model of ischemia and reperfusion in vivo]. AB - Rat adaptation to repeated periods of hypobaric hypoxia has been found to prevent the occurrence of ischemic and reperfusion ventricular arrhythmias on a 10-minte coronary artery occlusion model. Inhibition of delta-opioid receptors by intravenous administration of the selective delta-opioid antagonist TIPP (psi) in a dose of 0.5 mg/kg, intravenously (i.v.), completely abolished the antiarrhythmic effect of adaptation to hypoxia. Inhibition of mu-opioid receptors by CTAP (0.5 mg/kg, i.v.) or kappa-receptors by nor-binaltorphimine (9 mg/kg i.v.) had no effect on the incidence cardiac rhythm disturbances in adapted rats during coronary artery occlusion and reperfusion. Therefore, these findings suggest that delta-opioid receptors play an important role in inhibiting arrhythmia formation in this model. PMID- 12652939 TI - [Administration of the omega-3 polyunsaturated fatty acid drug eiferol decreases alcohol motivation in albino rats by elevating the level of antibodies to alcohol dehydrogenase]. AB - The study experimentally assessed the approach proposed by the authors to lower alcohol motivation, which involves enhancement of a specific immunity at the stage of alcoholization when acetaldehydemodified ethanol exchange enzymes [alcohol dehydrogenase (ADH)] and acetaldehyde dehydrogenase may be expected to occur. Omega-3 polyunsaturated fatty acid (PUFA) drugs enhance the formation of autoantibodies to modified ADH and decrease the activity of ADH in the stomach and liver. At the same time, PUFA drugs can, under certain conditions, produce an anti-alcoholic activity and a positive effect on the psychoemotional status of animals after the ethanol deprivation period. PMID- 12652940 TI - [The antioxidative defense system in the liver of rat fetuses upon antenatal exposure to lead]. AB - The activity of Zn, Cu-containing superoxide dismutase was studied in the liver of 20-day old fetuses of female rats exposed to different concentrations of lead in different periods and during pregnancy. There were significant changes in the enzyme activity depended on a used dose of lead and on the duration of exposure to this compound. Activation of free radical processes and impairment of the antioxidative defense system are likely to be one of the basic mechanisms responsible for hepatic damage during prolonged antenatal lead intoxication. PMID- 12652941 TI - [Potentiality of correction of the antioxidant enzyme system with electrically activated solutions]. AB - Electrochemically activated solution (ECAS) exerts effects on the activity of antioxidative enzymes (catalase, peroxidase, and superoxide dismutase), by causing an increase in the reduced activity of the enzymes and a reduction in their superactivity. The baseline activity of enzymes is subject to expressed individual variations in both animals and man. The cause of ECAS effects is, probably, the training of the antioxidative system due to the excess of electrons at the negative redox ECAS potential. PMID- 12652942 TI - [The lipid peroxidation system in the organism of experimental animals after exposure to cyclic hydrocarbons]. AB - The paper presents the results of studies of the amounts of diene conjugates, malonic dialdehyde, and Schiff's bases, which are molecular LPO products in the tissues of the lung, liver, kidney in animals exposed to 4-hour inhalation of cyclic hydrocarbons (pyromellitic dianhydride, durol, pseudocumene, and dioxane 1,4). Exposure to ecotoxicants at concentrations of 10 and 1 mg/m3 was found to result in the accumulation of initial, intermediate, and final molecular LPO products. These changes were recorded only after exposure to pseudocumene and dioxane vapors in a dose of 0.1 mg/m3 and after inhalation of dioxane-1,4 vapors in a dose of 0.01 mg/m3. PMID- 12652943 TI - [Altered renal function in acute sodium nitrite intoxication experiment]. AB - In experiments on non-inbred albino rats, intoxication was reproduced with 1% aqueous sodium nitrite solution, which was rated by the degree of hemic hypoxia (HH). The findings suggest that changes in renal functions (osmo-regulating, sodium-regulating) occur due to the development of mild HH, by increasing as hypoxia deteriorates. Decreased glomerular filtration and proteinuria, which are concomitantly detected, point to their mixed glomerulotubular pattern of renal lesion in nitrite intoxication. PMID- 12652944 TI - [Effect of lactoferrin on the development of acute and chronic adjuvant arthritis]. AB - An experiment revealed that the therapeutic administration of lactoferrin exerted no effect on the development of early stages of adjuvant arthritis in albino rats during chronicity. This was accompanied by decreased complement consumption and by the lower blood activity of polymorphonuclear leukocytes. The authors consider that this effect is associated with the capacity of lactoferrin to inhibit the excessive proinflammatory activity of neutrophilic leukocytes, which is characteristic of the chronicity of inflammation. PMID- 12652946 TI - [Modern concept of pathogenesis of cholelithiasis as a basis for principles of preventing biliary pathologies]. PMID- 12652947 TI - [Mitral valve prolapse in therapeutic and cardiology practice]. PMID- 12652945 TI - [The blood level of nitric oxide metabolites in experimental autoimmune disease during immunosuppressive therapy]. AB - Experimental autoimmune disease causes a drastic increase in the production of nitric oxide metabolites. The latter correlate with the indices of autoimmunity expression, with the levels of cyclic nucleotides, with the severity of morphological impairment of the structures of immunocompetent organs. The use of immunosuppressive agents in the pathogenetic therapy of an autoimmune process normalizes both the blood levels of nitrites/nitrates and immunological parameters; this reflects a clear-cut relationship between the synthesis of nitric oxide and the immune response of the body. PMID- 12652948 TI - [Constipation and principles of its treatment]. PMID- 12652949 TI - [Organization of hematologic work in a day hospital]. PMID- 12652950 TI - [Prevalence of bronchial asthma and allergic rhinitis in the adult population of St. Petersburg]. AB - AIM: To evaluate prevalence of bronchial asthma (BA) and allergic rhinitis (AR) among adult population of St-Petersburg. MATERIAL AND METHODS: Responders to ECRHS and B. Sibbald & E. Rink questionnaires who gave at least one positive answer were examined clinically, functionally and allergologically. A total of 3634 responders aged 16 to 98 years participated in the study. RESULTS: By the answers of the responders prevalence of BA was 7.2%, of AR--9.9%. Such figures were much higher than official statistics. Atopic BA was most frequent among other types--64%. CONCLUSION: It became evident that BA and AR in many cases are diagnosed late or are not recognized at all. This leads to growing number of patients with moderate and severe course of these diseases and deteriorates effectiveness of BA and AR treatment. PMID- 12652951 TI - [10-year dynamics of attitude to health problems in the male population of Novosibirsk (epidemiological study based on the WHO MONICA program)]. AB - AIM: To evaluate changes for a decade in the attitude of men in Novosibirsk to health problems. MATERIAL AND METHODS: WHO program MONICA has covered males aged 25-64 years (a representative sample from the population in one of the districts of Novosibirsk city). A total of 3 trials were made (in 1984, 1988 and 1994) which included questioning, registration of ECG, arterial pressure, height, body mass, biochemical tests of the blood. RESULTS: Attitude of men to their health depended on their age. There was a trend to evaluate their health as more and more poor in men at the age of 25-43 and 35-44 years. In the group of 45-54-year olds positive assessment of health was encountered 1.9 times more frequently, but the difference was not significant. At the age 55-64 years a growing number of men tend to assess their health as good. Since 1994 alcoholics among the elderly men grew in number as a response to the social and economic crisis. CONCLUSION: The change in health evaluation from negative to positive in older men may relate to less intensive work. PMID- 12652952 TI - [Current medico-ecological aspects of internal diseases in the urbanized north]. AB - AIM: To define a complex action of extreme climatic and harmful environmental factors of present-day northern industrial city on the course of chronic visceral diseases. MATERIAL AND METHODS: Dynamics of exacerbations of basic therapeutic nosologies was studied basing on the data on 10,169 inpatients treated in therapeutic departments of Surgut hospitals (Khanty-Mansi District) in 1994-1998. In parallel, ambient air of the city was controlled for 3 main climatic and 6 technogenic hazards. The analysis was made by means of medicoecological mapping of residential zones. RESULTS: A common feature of chronic visceral diseases course was determined: maximum number of recurrences in February-March and October-November, minimum number--in July-August. There is a relationship between the recurrences and extreme climatic factors. An increase in the number of exacerbations may be related with suppression of nonspecific resistance of the northerners' organism in these seasons. Differential study of the decompensation rate of chronic noninfectious diseases in residential zones in dependence with the degree of technogenic pollution has shown that areas of the town with high hospitalization rates are exposed to high mean annual pollution with sulfur dioxide, vanadium pentoxide and carbon oxide. Areas with low morbidity had the best environment. CONCLUSION: A combined action of extreme climatic and environment factors of the North plays an essential role in decompensation of chronic diseases in the northerners. Dynamics of recurrent chronic noninfectious diseases may serve a sensitive marker of ecological safety of the region. Design and realization of recreation and rehabilitation measures in ecologically polluted areas of the town are effective for prevention of aggravated chronic visceral diseases. PMID- 12652953 TI - [Nutrition of native rural population of the Russian north]. AB - AIM: To study actual nutrition of adult native population of the Russian North. MATERIAL AND METHODS: We have questioned 542 men and women living in Yakutia and Evenkia by 24-hour recall method (intake of the main food products). RESULTS: It was found that Yakutia and Evenkia population consume more proteins and fats than population of the European part of the RF and Central Siberia. 24-h energy requirements are provided with fats and carbohydrates. All the fats were animal. Simple sugars and cholesterol exceed recommended value. The quantity of the consumed fat and total cholesterol levels in blood serum correlated. Caloric value of the daily diet of Yakutia population is higher than that of Evenkia population. PMID- 12652954 TI - [Daily profile of arterial pressure in patients with arterial hypertension, continuously living in Tyumen polar region]. AB - AIM: To study a 24-h profile of arterial hypertension and affection of target organs in hypertensive population of the Tyumen North. MATERIAL AND METHODS: 62 patients (43 men and 19 women, age 18-50 years) with arterial hypertension (AH) living in the North of the Tyumen region entered the test group. The control group consisted of 56 AH patients (36 men and 20 women) matched by age. They lived in the zone of moderate climate (in Tyumen). 24-h arterial pressure (AP) monitoring was made starting on day 2 of the patients' arrival in Tyumen. Echocardiography was made on the arrival day. The examination of the groups was conducted against "clean" background. RESULTS: The groups did not differ significantly by age, sex, AH duration, office systolic and diastolic pressure (SP, DP). However, 37 patients from the test group on arrival in Tyumen showed normalization of both office AP and mean 24-h AP. 25 patients from this group sustained high office and mean 24-h AP. An abnormal 24-h AP profile with nocturnal hypertension was recorded in 76.6% of the test group and 28.31% patients of the control group. Left ventricular hypertrophy was significantly more pronounced in the test group than in the control one. CONCLUSION: In 2/3 patients from the North with AH stage 1 and 2 office AP and mean 24-h AP normalized upon their arrival to the zone of moderate climate but their left ventricular hypertrophy was more severe and changes in a 24-h AP profile with nocturnal hypertension persisted. PMID- 12652955 TI - [Various forms of mononitrates in treating patients with stress angina]. AB - AIM: Clinical and device evaluation of efficacy of new dosage forms of isosorbide 5-mononitrate (ISMN) in patients with angina of effort of functional class (FC) II-III in 1-3 month treatment. MATERIAL AND METHODS: 54 patients with ischemic heart disease (IHD) and stable angina pectoris (42 males, 12 females aged 48-70, mean age 59.4 +/- 3.6 years) were examined. 16 of them had angina of effort FC II, 35--of FC III and 3--of FC IV. IHD was verified at stress tests and 24-h Holter ECG monitoring. In 85.2% of IHD patients the diagnosis of stenotic coronary atherosclerosis of 1-3 great arteries was confirmed by coronarography findings. The treatment effect was assessed by bicycle exercise tests, Holter ECG monitoring, patients' records of anginal attacks before and after treatment. RESULTS: Long-term forms of ISMN--Mono Mac Depo (100 mg) and Mono Mac 50D (50 mg) -and non-retard form Mono Mac (40 mg) produced a stable antianginal effect in 85, 80 and 75% patients, respectively. ISMN preparations given for 1 and 3 months reduced frequency of anginal attacks, improved functional condition of the patients, exercise tolerance, variability of heart rhythm. Mono Mac Depo and Mono Mac 50 D were well tolerated, induced no serious side effects. CONCLUSION: ISMN preparations can be recommended as monotherapy in patients of FC II-III angina or in combination with beta adrenoblockers for treatment of patients with angina FC III-IV. PMID- 12652956 TI - [Effect of combined therapy with enalapril maleate and hypothiazide on the structuro-functional status of erythrocyte membranes in hypertensive patients]. AB - AIM: To characterize changes in erythrocytic membranes (ErM) structure and function in patients with essential hypertension (EH) due to combined therapy with enalapril maleate (EM) and hypothiazide. MATERIAL AND METHODS: Polypeptide and lipid spectra of ErM were studied by electrophoresis in polyacrylamide gel and thin-layer chromatography, respectively, the activity of transport ATP, lipid peroxidation (LPO) and antioxidant defense were studied with standard techniques in 32 EH patients before and 4 weeks after combined therapy with EM and hypothiazide. RESULTS: If previous EM therapy failed, combined treatment's effect was 62.5%. Molecular effects of the treatment consisted of decreased concentrations of actin and blend 5 protein, increased levels of spectrin in EM polypeptide composition, correction of phopholipid fractions imbalance, gluthationperoxidase. Transport ATP activity and chholesterin level remained unchanged. CONCLUSION: EH treatment with EM + hypothiazide is both clinically effective and beneficial for physico-chemical properties of the cell membranes. PMID- 12652957 TI - [Prognostic value of the degree of night decrease of systolic pressure in patients with mild and moderate forms of hypertension (7-9-year prospective study)]. AB - AIM: To study a relationship between a carcadian blood pressure (BP) rhythm and cardiovascular events (CE) during 7-9 year follow-up in males with mild to moderate essential hypertension (EH). MATERIAL AND METHODS: 50 males (mean age 48.6 +/- 0.7 years) with mild to moderate EH were prospectively followed up for 7 9 years (8.4 +/- 0.1 years). We analysed 24-h BP recordings and protocol of echocardiography performed during the first hospitalization. The patients were divided into three groups: group 1 (n = 18) with normal (10-20%) nocturnal fall of systolic BP (NF SBP) and normal left ventricular mass index (LVMI < 125 g/m2); group 2 (n = 16) with insufficient (< 10%) NF SBP and normal LVMI; group 3 (n = 16) with LVMI > 125 g/m2. In these groups we assessed the prevalence of CE: myocardial infarction (MI), stroke (S), sudden death (SD), new cases of angina pectoris (AP), transient cerebral ischemic attack (TIA). RESULTS: No significant differences were found between the groups by mean age, body mass index, duration of arterial hypertension, mean 24-h and awake systolic and diastolic BP while significant differences were by nighttime BP profile parameters. During the follow-up 16 CE in 12 patients were documented (3 fatal and 13 nonfatal). In group 1 CE were observed in 1 patient (twice MI), in group 2-7 cases of CE (1 S, 1 TCIA, 2 MI, 2 AP) in 6 patients, in group 3-7 cases (2 MI, 3 TIA, 2 AP) in 5 patients, 3 of them were fatal. CONCLUSION: Insufficient nocturnal fall of SBP (< 10%) is an adverse prognostic factor for cardiovascular morbidity in mild to moderate essential male hypertensives. PMID- 12652958 TI - [Angiotensin-converting enzyme, quinapril, in treating chronic cardiac failure]. AB - AIM: To assess effectiveness of ACE inhibitor quinapril in patients with moderate cardiac failure of ischemic etiology. MATERIAL AND METHODS: 20 patients received quinapril in a daily dose 20 mg for 6 months. Clinical condition, exercise tolerance, functional and geometric parameters of the left ventricle, Holter monitoring, heart rhythm variability, late potentials were studied before and after the treatment. RESULTS: The treatment relieved symptoms significantly, exercise tolerance increased, positive changes occurred in systotic, diastotic functions and left ventricular geometry. Ischemic episodes, supraventricular extrasystoles reduced in number and duration. CONCLUSION: Quinapril is an effective drug for treatment of moderate ischemic heart failure. PMID- 12652959 TI - [Efficacy of anti-inflammatory treatment with benacort in exacerbations of chronic obstructive lung disease]. AB - AIM: To determine a course and prognostic effectiveness of corticosteroid therapy of chronic obstructive pulmonary disease (COPD). MATERIAL AND METHODS: Long-term therapy with broncholytics (berodual) and benacort was given to 25 patients in non-infectious exacerbation, 20 patients received broncholitics of 45 COPD patients with mild and moderate ventilation disorders. Follow-up lasted for 2.5 years. Treatment effectiveness was judged by clinical indices, external respiration function, parameters of systemic immunity, cytogram of induced sputum and concentration of inflammatory markers in the sputum. RESULTS: An immediate effect of two-month therapy with benacort of COPD exacerbations manifested with positive clinical dynamics and relief of endobronchial inflammation. Prognostic efficacy--with a significant reduction of the frequency of COPD exacerbations and of their mean duration more than 3-fold. CONCLUSION: The results of long-term follow-up support validity of inhalation corticosteroids use for eradication and prevention of noninfectious exacerbations of COPD. PMID- 12652960 TI - [Significance of damage index in predicting systemic lupus erythematosus outcome]. AB - AIM: To determine the significance of the damage index (SLICC/ACR DI) in the prognosis of the outcome of systemic lupus erythematosus (SLE). MATERIALS AND METHODS: Correlations between the disease activity, cumulative damage and outcome were studied in 103 patients with SLE divided into two groups by the disease duration (21 to 60 months--group 1, 62 to 488 months--group 2). SLE activity was determined by Nasonova's classification, SLEDAI-1 and ECLAM. Cumulative damage was assessed by the damage index (systemic lupus international collaborating clinics/ACR damage index). RESULTS: We found the direct correlation between the age of the disease onset, SLE duration, degree of SLE activity and the damage index and between the maximal dose of glucocorticosteroids and the damage index. CONCLUSION: The results indicate that SLICC/ACR DI may be useful for outcome prediction in SLE. PMID- 12652961 TI - [Correction of respiratory organ impairment with Ural licorice preparations in chronic skin diseases]. AB - AIM: To study alterations in lipid peroxidation, antioxidant status of the lungs and immune state of the organism in some chronic diseases of the skin and potentialities of their correction with preparation produced of Ural licorice root. MATERIAL AND METHODS: Lipid peroxidation, antioxidant system parameters were examined in condensates of the expired air of 54 healthy subjects and 60 patients with chronic skin diseases. Immune status was studied in blood samples. The patients were given the drug made of Ural licorice root. RESULTS: The licorice-based drug normalized antioxidant defense and immune status. CONCLUSION: It was found possible to correct pulmonary and immune statuses of patients with some chronic skin diseases in polluted environment with a licorice drug. PMID- 12652962 TI - [Use of Mabtera (rituximab) in treating patients with refractory courses of B cell lymphoma, along with high-dose chemotherapy and autologous transplantation of hematopoietic stem cells]. AB - AIM: To study efficacy of rituximab in patients with resistant B-cell lymphoma on high-dose chemotherapy. MATERIAL AND METHODS: From September 2000 to April 2002 we studied efficacy and tolerance of rituximab at different stages of high-dose chemotherapy. The treatment was given to 10 patients with histologically verified CD20+ non-Hodgkin's lymphoma: diffuse large-cell (n = 4), Berkitt's (n = 2), follicular (n = 3), mantle-cell (n = 1). Five patients with diffuse large-cell lymphoma and Berkitt's lymphoma had a primary resistant course of the disease, one patient with diffuse large-cell lymphoma had a refractory recurrence. Follicular and mantle-cell lymphomas were characterized by a resistant course and large tumor masses. The patients received 1-2 courses of induction chemotherapy with dexa-BEAM with collection of peripheral stem cells followed by high-dose chemotherapy (BEAM-9, CBV + mitoxantron-1) with transplantation of autologous stem blood cells. Rituximab infusion (375 mg/m2) was conducted before the collection of the stem cells, prior to high-dose chemotherapy and in posttransplantation period after recovery of hemopoiesis. RESULTS: 4 patients achieved complete remission, 3-partial remission, 2 had progression and 1 stabilization. In mean follow-up 11 (2-20) months 7 of 10 patients were alive, overall survival being 15 +/- 2.4 months (95% confidence interval 10-19.7), median was not reached. 5 patients are in complete remission: 2 of them without further treatment, 3-after progression and repeat therapy including rituximab and interferon-alpha or rotuximab and CHOP chemotherapy. CONCLUSION: The addition of rituximab can improve the results of high-dose chemotherapy of patients with non Hodgkin's lymphoma resistant to standard doses of cytostatics. Repeat use of this drug can be effective in some patients with progression after high-dose chemotherapy with rituximab. PMID- 12652963 TI - [Transient myocardial ischemia in ischemic heart disease: diagnostic methods and treatment strategies]. PMID- 12652965 TI - [Changes in the skin in internal diseases (dermadromes)]. PMID- 12652964 TI - [Role of insulin resistance in pathogenesis of type 2 diabetes]. PMID- 12652966 TI - Laparoscopic intraperitoneal repair of postoperative ventral incisional hernia using Composix mesh. AB - This report describes the technique and early results obtained with a simple laparoscopic intraperitoneal onlay Composix mesh repair for postoperative ventral hernia. Composix mesh is constructed from one layer of polypropylene mesh and another layer of expanded polytetrafluoroethylene (ePTFE). From March 2000 to October 2001, we performed laparoscopic repair of postoperative ventral hernia in 9 patients. Four (44%) of these patients had a history of at least one failed hernia repair. The size of the abdominal wall defect varied from 4 x 5 cm to 10 x 12 cm (median, 8 x 9 cm). In all cases, the Composix mesh (Bard Inc. USA) was stapled to the peritoneal surface of the abdominal wall, leaving the sac in situ. No death occurred as a result of surgery. Intraoperative small bowel injury occurred in one patient (11.1%) for whom surgery was converted to laparotomy and small bowel resection. No infection was observed. The length of hospital stay varied from 5 to 10 days (median, 5.6 days). During the follow-up period of 8 to 15 months (median, 2 months), there was no recurrence of hernia. Laparoscopic Composix mesh onlay repair is a safe, easy, and effective procedure with minimal discomfort and a low early recurrence rate. PMID- 12652967 TI - Effects of TGF-beta and TGF-beta-neutralizing antibody on normal skin fibroblasts and scar-derived fibroblasts. AB - Transforming growth factor-beta (TGF-beta) is known as an important cytokine for scar formation in wound healing. The purpose of the present study was to investigate the effects of TGF-beta and its neutralizing antibody on normal skin fibroblasts and scar-derived fibroblasts in culture. Endogenous TGF-beta levels were similar in all fibroblasts. Cell proliferation increased when TGF-beta 1 or beta 2 was added to the cultures, and the increase was higher and started at a lower level in the scar-derived fibroblasts (p < 0.05). The increase of fibroblasts was suppressed by the addition of TGF-beta-neutralizing antibody to the cultures, and the suppression rate was higher in the scar-derived cells (p < 0.05). Percentages of the cells in the growth phases of cell cycle decreased in the normal skin fibroblasts (p < 0.05) when TGF-beta-neutralizing antibody was added. Our findings showed that scar-derived fibroblasts and normal skin fibroblasts have a different sensitivity to TGF-beta. Further study is needed on the effect of the neutralizing antibody to the cell counts and cell cycle of scar derived fibroblasts. PMID- 12652968 TI - Regional diminution of von Willebrand factor expression on the endothelial covering arachnoid granulations of human, monkey and dog brain. AB - Arachnoid granulation is a protrusion of the arachnoid membrane into the cranial sinus, and is thought to play an essential role in the cerebrospinal fluid (CSF) absorption. Because the cells covering the apex region of the arachnoid granulation have different morphological features compared to the ordinary endothelial cells lining of the cranial sinus lumen, it has been expected these covering endothelial cells perform some specific function in the CSF absorption mechanism. However, little is known about functional differences between the covering endothelium of the arachnoid granulation and the ordinary sinus endothelium. In the present study, the characteristics of the covering cells located at the apex of arachnoid granulations of human, monkey and dog brain were examined by histochemical and immunohistochemical methods. The endothelial cells lining the cranial sinus lumen generally expressed such proteins as von Willebrand factor (vWF), CD31 and glycoproteins containing GS-1 or LE-1 lectin reacting sugar residue which are endothelial cell markers. However, the endothelial cells specifically located at the apex of arachnoid granulations failed to show vWF immunoreactivity, whereas the other endothelial markers were positive in each species we examined. Double staining of vWF antibody with other markers has clearly demonstrated that the endothelial cells on the apex region of arachnoid granulations exhibit no expression of vWF whereas cells lining the lateral region of arachnoid granulations and the luminal surface of ordinary cranial sinuses showed co-localization of these markers. The structural and histochemical differences between endothelial cells located at the apex region of arachnoid granulations and those of the sinus wall may reflect functional differences. PMID- 12652969 TI - Effects of methylphenidate on the inhibitory postsynaptic potential in rat locus coeruleus neurons. AB - Effects of methylphenidate (MPH), an agent used clinically for the treatment of children presenting the attention-deficit/hyperactivity disorder (AD/HD), on synaptic transmission in the rat locus coeruleus (LC) were examined by intracellular recording methods. Bath-application of MPH (30 nM-3 microM) increased the amplitude of the inhibitory postsynaptic potential (IPSP), while it did not change the amplitude of the excitatory postsynaptic potential (EPSP). MPH increased the time-to-peak and the half-decay time of the IPSP in LC neurons. MPH increased the amplitude of spontaneous IPSP: individual spontaneous IPSPs merged one into the other so as to produce regular, long-lasting waves of hyperpolarization. Clonidine (10 nM), a selective agonist for alpha 2 adrenoceptors, depressed the IPSP without affecting the EPSP in LC neurons. The results suggest that MPH enhances inhibitory synaptic transmission in the rat LC by depressing the norepinephrine (NE) re-uptake system. PMID- 12652970 TI - Effects of supplemental sialyl Lewis(x) analogue during warm blood cardioplegia. AB - Effects of supplemental Sialyl Lewisx analogue, a major ligand for all three selectin family members, during warm blood cardioplegia were assessed in the blood perfused isolated rat heart. The isolated hearts were arrested for 60 min with warm blood cardioplegia given at 20-min intervals. This was followed by 60 min of reperfusion. The hearts were divided into the following two groups according to the supplemental drugs added to the cardioplegic solution. The control group (n = 6) received standard warm blood cardioplegia. The Sialyl Lewisx analogue group (n = 6) received warm blood cardioplegia supplemented with Sialyl Lewisx analogue (60 micrograms/ml). Cardiac function, endothelial function, myocardial metabolism and myocardial myeloperoxidase activity were assessed before and after cardioplegic arrest. Left ventricular developed pressure and dp/dt were significantly (p < 0.05) greater and -dp/dt was significantly (p < 0.05) lower in the Sialyl Lewisx analogue group than the control group during reperfusion. Coronary flow at 15 min of reperfusion and NO production, when acetylcholine chloride was added were significantly (p < 0.05) greater in the Sialyl Lewisx analogue group than the control group. Myeloperoxidase activity was significantly (p < 0.05) lower in the Sialyl Lewisx analogue group than the control group. The results suggest that selectin-mediated endothelial-leukocyte interactions may play an important role in myocardial ischemia and reperfusion injury. Supplementation of Sialyl Lewisx analogue during warm blood cardioplegia may provide superior myocardial protection by suppressing leukocyte-endothelial interaction during early reperfusion period. PMID- 12652971 TI - Role of heparan sulfate proteoglycan (syndecan-1) on the renal epithelial cells during calcium oxalate monohydrate crystal attachment. AB - We have reported that heparan sulfate (HS)/heparan sulfate proteoglycan (HSPG, syndecan-1) expression significantly increased in the rat kidney during calcium oxalate (CaOx) nephrolithiasis. Although the exact role of syndecan still remains unclear, HS/syndecan-1 is thought to have some important role in the CaOx crystal formation. Mardin-Darby canine kidney (MDCK) cells are most commonly used in kidney stone research. It was reported that MDCK cells do not express syndecan-1. In the present study, we established a novel MDCK cell line (KIC-synd-1) that expressed the human syndecan-1 gene. In this cell line, we confirmed stable expression of both sndecan-1 gene and core protein. Immunohistochemical study revealed positive staining of syndecan-1 monoclonal antibody in the basolateral and cytosolic area of the KIC-synd-1 cells. We also investigated the composition of glycosaminoglycans (GAGs) side chains in MDCK cells and KIC-synd-1 cells by using high-performance liquid chromatography (HPLC). Four types of HS chains were identified in both cells as follows; delta diHS-NS, delta diHS-6S, delta diHS diS1, delta diHS-diS2. Increased production of delta diHS-NS and delta diHS-diS2 were shown in KIC-synd-1 cells compared with production in MDCK cells (p < 0.05). In contrast, only a small amount of delta diHS-6S and delta diHS-diS1 was contained in both cell lines. Total amount of HS was significantly increased in the KIC-synd-1 cells compare with that in the wild type MDCK cells (p < 0.05). Scanning electron microscopy revealed no significant difference between cell surface of wild type MDCK cells and that of KIC-synd-1 cells in normal conditions. However, calcium oxalate crystal attachment was apparently decreased in the KIC-synd-1 cells. These results suggested that cell surface HS/syndecan-1 has preventive role for calcium oxalate nephrolithiasis via creation of a charge barrier against COM crystal attachment. PMID- 12652972 TI - Antiphospholipid syndrome and multiple ischemic strokes in a patient with myasthenia gravis. AB - Little is known about ischemic stroke occurrence in patients with myasthenia gravis (MG), although antiphospholipid antibodies are detectable in many MG patients. A 47-year-old woman with a 20-year history of generalized MG had an acute onset of right hemiparesis. She had undergone thymectomy 10 years previously and was treated for phlebothrombosis of the lower extremity 3 years previously. Computed tomography (CT) demonstrated an old infarct in the left frontal lobe and a new lesion in the right parietal lobe. Multiple small cortical and subcortical infarcts were demonstrated on fluid attenuated inversion recovery (FLAIR) images. Thrombocytopenia (5.9 x 10(4)/microL), a prolonged activated partial thromboplastin time (aPTT; 50.2 sec), and an elevation of beta 2-IgG glycoprotein I anticardiolipin antibodies (beta 2-GPIaCL; 55.7 U/mL) were observed. Neurological defects improved significantly over 2 weeks. She then was treated with oral prednisolone (30 mg/day) for 18 months, with resolution of laboratory abnormalities and no new cerebrovascular events or findings on imaging. We believe that our patient's multiple infarcts are caused by antiphospholipid antibodies and recommend glucocorticoid therapy to prevent recurrent of ischemic stroke in similar case. PMID- 12652973 TI - Appendicitis caused by caecal carcinoma: report of a case. AB - Appendicitis can rarely occur in association with carcinoma of the caecum, particularly in elderly patients. We report a case of acute appendicitis provoked by an adenocarcinoma of the caecum which obstructs the lumen of the appendix in a 58-year-old man. The patient underwent an ileocaecal resection with lymph node dissection. The difficulties of identifying a small tumor at laparotomy and the implication for optimal treatment are emphasized. PMID- 12652974 TI - [Impact of exogenous infection on tuberculosis infection rates in children and adolescents]. AB - The impact of exogenous infection on the course of an epidemic process was considered. The spread of tuberculosis infection from an ill patient to children and adolescents within a territorial focus was observed. A mapping study allowed the authors to make a goal-oriented search for patients with tuberculosis, as well as primarily infected children and adolescents. PMID- 12652975 TI - [Prevalence of tuberculosis and its specific clinical features in children]. AB - A retrospective study of epidemiological indices of tuberculosis in children in the Republic of Uzbekistan for the past 20 years has shown that tuberculosis morbidity rates have increased by 65.4% and by 114% in children living in the foci of tuberculosis infection. The greatest increase in its morbidity rates is observed in the Aral Sea region, which is accompanied by worse social, environmental, and economic situations. Varying tuberculosis courses are revealed in inland children, which makes it necessary to implement tuberculosis controlling measures, especially in the foci of tuberculosis infection. PMID- 12652976 TI - [Antituberculous measures according to the results of Mantoux test]. AB - Analyzing the results of Mantoux tests with 2 TE made 7 times for 5 years in 750 850 schoolchildren of the settlement of Sulak has demonstrated that the infection rates are on the increase on the one hand, and, due to material and transport problems, the proportion of examinees from a risk group decreases to 10-30% of those to be examined, on the other hand. The authors suggest that the remaining unexamined children should undergo chemotherapy with 2-3 antituberculosis drugs for 3 months so as to perform an examination when the first opportunity occurs. They propose to examine the coverage of children, except for children and adolescents with a hyperergic and increasing reactions and those with a highly positive (15-16 mm) Mantoux test. An algorithm for implementation of the above measures is offered. PMID- 12652977 TI - [Specific features of tuberculin sensitivity in children with allergic dermatoses]. AB - The recent rise in the incidence of allergic diseases, including allergic dermatoses, requires a comprehensive approach to solving a number of problems associated with this disease. Analyzing the case reports of 62 children with allergic dermatoses has indicated that this is a particular contingent of children, which needs a more careful approach to different preventive measures, to interpreting tuberculin reactions and hence to substantiating the use chemotherapeutical prevention. A graded skin tuberculin test and an immunological study should be performed in critical cases. PMID- 12652978 TI - [New technologies in the prevention, detection, diagnosis and treatment of tuberculosis in children]. AB - If the epidemiological situation is tense, new technologies should be developed and put into practice to enhance the efficiency of specific prevention, early detection, diagnosis and treatment of tuberculosis in children. There is evidence for the high efficacy and low reactogenicity of lower antigenicity-loading BCG-M vaccine that causes a 15-fold decrease in infant morbidity, as compared with that among non-vaccinated children, and this vaccine shows a 5-fold reduction in postvaccination complications as compared with BGC vaccine. The 26-year use of tuberculin diagnosis via Mantoux test with 2TE PPD-L during mass vaccination of children and adolescents has proved itself in early identification of tuberculosis and risk groups. A new risk group has been identified. This includes children with increasing tuberculin reactions; three-month intermittent chemoprevention with isoniazid reduces tuberculin sensitivity in the children and prevents tuberculosis in them. The developed short-term (6-9 months) courses of chemotherapy in preschool and school children by using drugs (isoniazid + rifampicin + pyrazinamide) yield the best healing without residual changes of uncomplicated forms of tuberculosis in 83 and 60% of the children with complicated events, respectively. The chemotherapy regimens have been divided into 4 groups of different dosage schedules. PMID- 12652979 TI - [Drug resistant tuberculosis in children and adolescents]. AB - The paper presents the results of a follow-up of children and adolescents who isolate drug-resistant Mycobacterium tuberculosis (MBT) strains of those with manifestations of tuberculosis and preserved sensitivity to antibacterial agents. Bacterial isolation in children and adolescents is scanty and multiple. There is a high rate of resistance to isoniazid, streptomycin, and rifampicin. The vast majority of children and adolescents who isolate drug-resistant MBT strains have been found to contact a patient who also isolate drug-resistant MBT. The drug sensitivity of MBT in patients is identical to that of sources of their contamination. Most patients with drug-resistant tuberculosis have been ascertained as having been registered in dispensaries and treated with drugs, which suggests that the use of chemotherapy in prophylactic doses, particularly in patients contacting a bacterial isolator, does not prevent local tuberculosis. PMID- 12652981 TI - [Specific features of tuberculosis in infants]. AB - Examining active tuberculosis in 38 infants has established the following features: the disease is characterized by predominantly (71%) classical forms of primary tuberculosis; moreover, a primary tuberculosis complex is more common (41.7%) than tuberculosis of intrathoracic lymph nodes (26.3%); by complicated tuberculosis in more than a third of the infants, by extrapulmonary tuberculosis in 23.7%, and by the affliction of 2 systems in 13.2%. Tuberculosis runs most severely in children under 1 year of age. The group at the highest risk for tuberculosis may include infants contacting patients with tuberculosis (65.8%), ill vaccinated infants with BCG (89.4%), infants with symptoms of intoxication and thoracic complaints (79.3%), and those with hyperergic and pronounced sensitivity to tuberculin. PMID- 12652980 TI - [Incidence of adverse reactions to chemotherapy and their types in adolescents with tuberculosis]. AB - During treatment, 65.7% of the patients developed adverse reactions to antituberculosis drugs. Allergic reactions were most commonly associated with the use of streptomycin. The simultaneous inclusion of isoniazid, rifampicin, and pyrazinamide into the treatment regimens irrespective of the total number of drugs was found to increase the incidence of hepatotoxic reactions as compared with the regimens which do not contain a combination of these 3 drugs. Cycloserin may be given to adolescents with multidrug resistant tuberculosis. Glutaminic acid prevents the toxic effect of the drug on the CNS, therefore cycloserin may be prescribed for a long period (for as long as 9 months). PMID- 12652982 TI - [Clinical characteristics of neonatal infants born to mothers with pulmonary tuberculosis]. AB - Three hundred and seventy neonatal infants born by mothers with pulmonary tuberculosis were examined. A control group comprised 121 neonatal babies born by apparently healthy women. The infants born by ill mothers weighed less. In the study group, 142 (38.4%) babies with asphyxia were born. In infants born by mothers with active tuberculosis, birth asphyxia was observed 1.5 times as frequently as in those born by healthy mothers. In the control group, only 11 (9.1%) children with mild asphyxia were born. Neonatal compilations were noted in 137 (37.1%) infants of the study group, moreover almost equally frequently in babies born by mothers with both active and inactive tuberculosis, they were in 12 (9.9%) babies in the control group. In the latter group, malformations were detected only in 2 (1.7%) newborn babies, in the study one, they were in 46 (12.4%) infants, the mothers of these 15 (32.6%) infants had received specific therapy for tuberculosis in early pregnancy. PMID- 12652983 TI - [Diagnosis and treatment of tuberculosis of peripheral lymph nodes in children and adolescents]. AB - Among lymphadenopathies of different etiology, tuberculosis of peripheral lymph nodes was detected in 28% of cases in children and adolescents, as shown by our data. In its diagnosis in the children, the most informative methods include a morphological study of lymph node biopsy specimens and the results of experimental treatment. History data, clinical findings, and tuberculin diagnosis were also of great importance in verifying the disease. Lymphotropic administration of antituberculosis agents in patients with tuberculosis of peripheral lymph nodes enhances the efficiency of chemotherapy. PMID- 12652984 TI - [Clinical and epidemiological characteristics of renal tuberculosis in children and adolescents]. AB - The clinical manifestations of renal tuberculosis under the present conditions were studied. Seventy one patients with active renal tuberculosis treated at the Moscow Research Institute of Phthisiopulmonology and at the Pediatric Department, Yekaterinburg Research- and Production Association Phthisiology were followed up. The morbidity rate of urinary tuberculosis was found to directly depends on whether early detection of the disease is organized. Urinary tuberculosis afflicts children and adolescents in all age groups. However, its higher rates are detected in the first decade of life. Renal tuberculosis is accompanied by manifestations of the urinary syndrome in 70.4% of cases and by the presence of Mycobacteria tuberculosis in 100%. A limited renal tuberculous process was detected in 80% of cases in the period of primary infection or in the following 2 years. A generalized renal tuberculous process involving other systems in a child was accompanied by few clinical urinary symptoms and it did not depend on the duration of infection. PMID- 12652985 TI - [Specific features of functional activity of neutrophilic granulocytes in infection with Mycobacterium tuberculosis in children of different age groups]. AB - Sixty children infected with Mycobacteria tuberculosis from two age groups (3-5 and 12-14 years) were followed up. The phagocytic activity and oxygen-dependent biocidicity of peripheral neutrophilic granulocytes were studied. The studies indicated dysfunctions of neutrophilic granulocytes in the children infected with Mycobacteria tuberculosis. This fact should be borne in mind in developing immunorehabilitative programmes in this category of patients. Moreover, the changes in the killing activity of neutrophilic turned out to be more pronounced in a young age group. In this connection, immunomodulating therapy should be strictly differentiated. PMID- 12652986 TI - [Effects of immunomodulator leukinferon on the course of experimental tuberculosis]. AB - An experiment was conducted on 60 CBA mice intravenously inoculated with cultured Mycobacteria tuberculosis (MBT), Erdmann strain, in a dose of 0.025 mg. The specific features of tissue, cellular, and biochemical reactions were studied in the lung, liver, and spleen when leukinferon (LF) was included into tuberculosis treatment regimen. LF was shown to have a positive impact on the development of reparative reactions during tuberculous inflammation by reducing the time of abacillation and recovering the structure of diseased organs. By month 3 of follow-up, MBT were not detected in mice receiving antibacterial agents (ABA) and LF, while typical and changed forms of LF were identified in the cytoplasm of alveolar macrophages in mice treated with ABA alone. A specific feature of an inflammatory reaction as a significant proliferation of lymphocytes and macrophages with their ample infiltration of target organs was noted in animals receiving LF. This was followed by the activated production of alpha- and gamma interferons and by the mobilization of an enzymatic link of anti-oxidant defense under chronic oxidative stress, which led to a reduction of resolution of inflammatory areas and to an increase in survival of animals which had not been given ABA, but treated with LF alone. PMID- 12652987 TI - [Some parameters of bronchoalveolar lavages in alveolitis]. AB - Eighty nine patients with alveolitis [60 with extrinsic allergic alveolitis (EAA) and 29 with idiopathic fibrosing alveolitis (IFA)] were followed up. Cytological and immunological studies of bronchoalveolar lavage revealed that the patients with EAA had elevated counts of lymphocytes, moderately increased neutrophils and eosinophils, decreased alveolar macrophages, elevated SIgA and T lymphocytes. In the patients with IFA, only higher counts of neutrophils were significant. PMID- 12652988 TI - [Examining the distribution of HLA antigens in patients with tuberculosis and healthy individuals in some regions of the Republic of Tyva]. AB - Healthy individuals and patients with pulmonary tuberculosis were typed by HLA antigens A, B, and C. They were all Tyvians and lived in different regions of the Republic of Tyva. Examination was made in 128 and 85 healthy individuals and 62 and 55 patients with pulmonary tuberculosis from the Todzhinsky and Erzinsky districts of the Republic of Tyva, respectively. The healthy populations of the districts in question were found to differ in a number of Class 1 HLA antigens, such as A3, A10, A11, A28, B7, B27, Cw4. In the Todzhinsky district, tuberculosis morbidity rates were also ascertained to be positively associated with the HLA antigen B13 whereas in the Erzinsky district they were associated with the HLA antigen B12. The findings may be used to form tuberculosis-risk groups in the study districts. PMID- 12652989 TI - [Mechanisms of large artery remodeling]. AB - Degenerative changes of elastin, increase of collagen fibres, proliferation of muscle cells and impairment in endothelium function occurs in the vessel wall with ageing, in hypertensive and others diseases. Early changes are potentially reversible. Atherosclerosis is a late stage of pathologies occurring in the arterial wall in response to various factors. The present paper describes current knowledge on mechanisms of arterial wall remodelling. The authors define basic indices of arterial compliance and describe several physiological and pathological factors which occur play the main role in process of arterial wall remodelling. A chapter is devoted to the effects of increased arterial stiffness on heart pathology and clinical implications of measuring these parameters. PMID- 12652990 TI - [Pulse wave analysis: from the basic sciences to clinical applications]. AB - Assessment of the pulse character is one of the earliest recorded medical skills, but objective recordings of pulse waveform emerged only in the nineteenth century. This technique fell into disuse with the advent of the sphygmomanometer, but interest has recently been rekindled with the introduction of computer technology and high fidelity applanation tonometry. Sphygmocardiography is a non invasive and accurate technology which utilizes tonometric analysis of the radial or carotid artery pulse wave and measurement of peripheral arterial blood pressure to derive a central arterial pulse wave, central arterial blood pressure and related indices. In clinical studies pulse wave analysis is a highly reproducible method and easy to apply. This measure provides important information about arterial stiffness and heart-vascular interactions. This technique may provides better cardiovascular risk assessment associated with increased arterial stiffness and allow treatment to be targeted to those most in need. PMID- 12652991 TI - [Vascular remodeling and the risk of cardiovascular events]. AB - Structural and functional changes of arterial walls are one of the underlying causes of many disorders which modern cardiology deals with. Information about arterial structure and function can be now obtained from various non-invasive vascular imaging tests, such as: measurements of carotid intima media thickness, assessment of biomechanical parameters of blood flow, endothelial function or coronary artery calcification. Despite the development of new diagnostic tools, the diagnosis of cardiovascular risk is still very difficult. This review discusses the role of arterial remodeling in the assessment of cardiovascular risk. Evidences presented in this paper demonstrate that the data about arterial function and structure can be used when deciding about the initiation of therapy and can provide information about the efficacy of treatment. PMID- 12652992 TI - [The therapeutic possibilities of the regression of the vascular changes in hypertension]. AB - Arterial hypertension has an important functional and morphological influence on the arterial system. The evaluation of functional and morphological changes is based upon such terms as compliance, stiffness, distensibility in the former and intima-media thickness (IMT), number and kind of atheromatous plaques in the latter case. Functional changes in the large arteries can be modified by antihypertensive and hypolipemic agents. The IMT as a non-invasive marker of vascular wall injury is increasingly frequently used in the studies. It has been identified as a significant predictor of coronary and cerebral vascular complications. Most studies on hypolipemic therapy demonstrate that the progression of IMT is slowed down. Ca-antagonists, ACE inhibitors and beta blockers are the most frequently studied agents with respect to their influence on IMT in the following patient populations: hypertensives, patients with coronary artery disease, at high cardiovascular risk and asymptomatic subjects with carotid plaque. The present paper summarises the results of these studies. The findings of the ELSA trial, the largest study evaluating the effects of therapy on IMT are noteworthy. It has been demonstrated that lacidipine has a better antiatheromatous effect than atenolol, which is independent of its hypotensive properties. Non-invasive, relatively easy ultrasound measurement of IMT in the carotid artery is an important tool to detect atheromatous lesions and because of its prognostic value. It may also serve as a predictor of cardiovascular complications, which helps in the process of therapeutic decision making. PMID- 12652993 TI - Prevention of maternal mortality: why success eludes us? PMID- 12652994 TI - AIDS awareness among undergraduate students, Maharashtra. AB - Total 313 undergraduate students (132 males and 181 females) of the colleges of Nashik and Talegaon of Maharashtra were surveyed regard awareness about AIDS. Most of the students knew about AIDs and tests available. They were aware that people indulging sexually promiscuous relations are at risk of AIDS. But the fact that it is transmitted by infected blood and from infected mother to child was not widely known, particularly among Arts students. Some misconceptions regarding modes of transmission were observed among few students, like social kissing, sharing utensils/personal items, using common swimming pools and insect bite spread infection. Attitude towards HIV infected/AIDS patients were not sympathetic. Overall knowledge of Science students were better compared to commerce and Arts students. Confusion about mode of transmission and prevention of the disease exist. Scope of health education for these students was well felt. PMID- 12652995 TI - A study on child care practices and school education in rural areas of Vijayanagaram district, A.P. AB - In Vijayanagaram District Survey a sample of 4800 rural population in 1070 households were contacted to study various child health indicators. 425 under five children and 971 children in 5-14 age groups were studied to estimate various child health key indicators. PMID- 12652996 TI - Women empowerment. PMID- 12652997 TI - Comparison of hemoglobin values obtained by HaemoCue and Sahli's methods. PMID- 12652998 TI - Drug inventory management in Medical College Hospital, Kolkata. PMID- 12652999 TI - Tuberculosis: discovery to DOTS. PMID- 12653000 TI - Health profile of lock factory workers in Aligarh. AB - In order to assess the health problems and its relationship to the nature of work of lock factory workers, a field based cross sectional study was carried out in sixty randomly selected lock factories in urban industrial areas of Aligarh city. Nine hundred fifty seven lock factory workers were included in the study out of them 45.7 percent were found to be suffering from one or more than one diseases. Detailed study of the workers showed that 73.05 percent were suffering from respiratory tract diseases, a majority of them having upper respiratory tract infection. Diseases showed a definite trend either increasing (Chronic bronchitis, Emphysema etc) or decreasing (upper respiratory tract infection, Bronchial asthma, Injuries & accidents), according to the age and duration of work. Polishing & filing appeared to be the most hazardous type of the work as shown in the proportion of chest diseases, (56.6%) & (38.3%) in polishing & filling workers respectively. PMID- 12653001 TI - The role of hearing aids in providing environmental feedback for the profoundly hearing impaired. AB - A number of studies have brought out the limitations of hearing aids in the rehabilitation of hearing impaired. This study was undertaken to assess the use of hearing aids in improving the ability of the hearing impaired to hear vehicular horns. The students of a school for the hearing impaired were interviewed with the help of their teachers. Out of total 102 study subjects, 47 (46%) said that they drove vehicles. None of them used hearing aid while driving vehicles. However, 56 (55%) students possessed functional box type hearing aids. Their ability to hear the sound of vehicular horns with and without hearing aids in the presence of background noise under experimental set up was assessed. None of the 36 study subjects could hear the horns (< 85 dB and > 85 dB) without hearing aid. Only 5 (16.6%) of the 30 profoundly hearing impaired students could hear the sound of horns (> 85 dB) with the hearing aid. The hearing aid was found to be more beneficial to subjects with severe hearing impairment. There is a need to bring forth newer and better varieties of hearing aids and other technologies e.g. Cochlear implant within the reach of those in need. PMID- 12653002 TI - Health care options in childhood ARI before hospital care. AB - A cross sectional study was conducted, to examine the pattern of health care options, exercised before seeking care at the district hospital in the event of ARI in under fives. One hundred fifteen under fives selected through systematic sampling technique, from two district hospitals were the subjects of study. Respondents were the care providers, who accompanied children to the Hospitals. Time delay in initiating care and reaching the district hospital was also recorded. Reasons for preferring a particular source as first choice were enquired. District Hospitals stood out as the most preferred source, as 52 (45.2%) of the children used it as the first step. Other sources of health care were Health Center (10.4%), Home Care (25.2%) General Practitioners (10.4%) and Drug Stores (8.7%). On an average a child took 1.8 steps before coming to the district hospital. Children experienced 13 unique treatment patterns. Children initially offered home care followed longer sequence and more variable pattern. Convenience (62.6%) and cost (37.4%) were the main factors in choosing a source of treatment. Children who received home care were brought to district hospital earlier than others. PMID- 12653004 TI - Clinical examination and hematological data in asymptomatic & apparently healthy school children in a boarding school in a tribal area. AB - In a boarding school of Maharashtra State of India 314 students (Bhil & Pawar) were examined clinically and blood was examined. Anemia was present in 16.2% male & 38.3% female. B (Beta). Thalasemia trait was present in 1.6% male & 2.4% female. Sickle cell trait was present in 21.3% male and 14.4% female and sickle cell disease in 0.6% student. G6PD deficiency was seen in 5.1% male & 4.8% female students. PMID- 12653003 TI - An evaluation of bed management in a rural hospital adjacent to Indo-Nepal border in West Bengal. AB - Bed management is one of the important activities for efficient hospital management. The present study on evaluation of bed management in a rural hospital revealed that the total bed capacity could not be utilised. The turnover rate, turnover interval, bed occupancy rate and average length of stay were closely corroborating. PMID- 12653005 TI - Immunization strengthening project. PMID- 12653006 TI - Reproductive and child health programme. AB - Reproductive and Child Health Programme is an integrated and comprehensive programme based on realistic decentralised area specific microplanning tailored to meet the local needs. There is adequate emphasis on provision of good quality of services under the programme. The present paper traces the genesis and evolution of the RCH programme followed by its objectives, strategies and components. Various mechanisms for the implementation, monitoring and evaluation of the programme are discussed. The programme is analysed critically and suggestions for improvement are provided. PMID- 12653008 TI - Overview of reproductive and child health programme. PMID- 12653009 TI - Safe motherhood and child survival. PMID- 12653010 TI - Nutrition. PMID- 12653007 TI - An overview of multicentric training workshops for public health professionals on reproductive and child health programme in India. AB - The major emphasis of Reproductive and Child Health (RCH) programme in India is delivery of client-oriented, demand driven and broader ranges of high quality, safe and effective services for children, adolescents, mothers and reproductive age group population at large. Increased client satisfaction is considered as main determinant for improved acceptance of the services. Thus, well trained and motivated health personnel are necessary to deal with highly sensitive, personal health issues of the clients, like contraception, abortion, infertility services etc. The Indian Public Health Association organized total 10 workshops in several places of India (A total 322 members, composed of Medical Administrators (54.7%), Faculty members of Medical Colleges (24.5%), Sociologist and Nutritionists (13.9%) and also public health personnel (6.9%) participated in the workshop). Learning objectives and lesson plans etc. were formulated. Accordingly the contents were incorporated in a module, validated and pretested. The training sessions were conducted by briefing, discussion, group exercise and VIPP method and were evaluated by semi structured. The pre/post assessment schedule and scored scale of feedback from participantsAE were used for evaluation. The pre post assessment scores revealed wide variations of mean score among the participants of several places. Significant post workshop improvement of knowledge was quite evident, with few exceptions. The group variants of pre and post score of results was considered to be due to heterogeneous groups of participants. Review of objectivity and quality of the questionnaire were felt as necessary. PMID- 12653012 TI - Prevention and management of reproductive tract infections/sexually transmitted infections. PMID- 12653011 TI - Adolescent health. PMID- 12653013 TI - Communication for behaviour change. PMID- 12653014 TI - Community need assessment & participatory learning for action. PMID- 12653015 TI - Dr. B. C. Das Guptta Memorial Oration. Challenges before public health organisations national health programmes. PMID- 12653016 TI - Dr. A. L. Saha Memorial Oration. Public health in India: future challenges. PMID- 12653017 TI - Optimal infant feeding in rural areas--the missing agenda of communication needs. PMID- 12653018 TI - Factors associated with detection of bromoxynil in a sample of rural residents. AB - In regions of intensive crop production residents may be exposed to herbicides through direct contact or environmental sources. The environmental herbicide exposures of rural populations and resultant potential health effects are not well understood. Epidemiologic studies or herbicides have focused on occupational exposures using, primarily, self-reported data (e.g., information on occupational and non-occupational herbicide use, agricultural practices and exposures, farm residence). Herbicide exposure characterization in epidemiologic research would be strengthened by the use of self-reported data and biological monitoring (e.g. measuring the herbicide parent compound or its metabolites in blood or urine specimens) to classify individual exposures, identify factors associated with exposure, and obtain integrated estimates of exposure. As both exposure metrics are susceptible to measurement error and some self-reported and biological monitoring data might not be correlated, a worthwhile first step is to identify self-reported data that are statistically associated with biological measures or exposure. This study use gas chromatography/mass spectrometry analysis to measure blood plasma concentrations of target herbicides in a sample of rural residents (men, women, and youths) of Saskatchewan, Canada, and identified factors, based on self-reported data, associated with detection. The questionnaire data and blood specimens were collected in February/March 1996 during winter (frozen soil and water and snow cover) conditions. Sixty-four of the 332 study participants (19.3%) had detectable levels of the herbicide bromoxynil although herbicide application in the region had not occurred for approximately 5 mo and bromoxynil has a relatively short environmental half-life. The prevalence of detection of other target herbicides (2,4-D, triallate, trifluralin, dicamba, fenoxaprop, MCPA, and ethalfluralin) varied from 0.3% to 2.7%. Self reported factors identified in the multiple-variable analysis as statistically significant predictors of bromoxynil detection included recent exposure to grain production as the main farming operation (statistically significant for producers and for non-farming family members of producers), a history of bromoxynil use, a history of having felt ill with a pesticide exposure and a history of pesticide spill on skin or clothing, with apparent gender differences in the relative importance of these factors. Detection of bromoxynil in this rural sample, 3-4 mo after freeze-up and winter snow cover, suggests either that bromoxynil is very slowly metabolized/excreted from the body or study participants were environmentally or occupationally exposed tothe herbicide during this period. Further research is needed to elucidate the pathways of exposure, biological half-life, and potentialhuman health effects of bromoxynil. PMID- 12653020 TI - Diesel exhaust particle-induced alterations of pulmonary phase I and phase II enzymes of rats. AB - Although diesel exhaust particles (DEP) are known to produce pulmonary disorders, the xenobiotic metabolic pathways associated with DEP detoxification and bioactivation remain unclear. In this study, the effect of acute exposure of DEP on phase I and phase II enzymes of rat lung was investigated. Intratracheal administration of DEP produced an induction of cytochrome P-450 (CYP) 1A1 enzyme protein and activity at 1 d postexposure, with the enzyme level returning to control at 5 d postexposure. On the other hand, carbon black (CB), a particle control, did not show any induction of CYP1A1 protein or enzyme activity. However, both DEP and CB significantly decreased CYP2B1 protein and enzyme activity at 1 d postexposure. The decrease in CYP2B1 enzyme protein and activity by DEP or CB treatment was observed up to 7 d postexposure. DEP and CB treatments also significantly attenuated glutathione S-transferase (GST)-pi protein at 1 d postexposure. Both DEP and CB at 35 mg/kg significantly decreased the activities of GST and catalase at 1 and 7 d postexposure. DEP, but not CB, significantly induced quinone reductase (QR) activity at 7 d postexposure. This study suggests that DEP may induce CYP1A1 and QR enzymes via a chemical effect, while the carbonaceous core may be involved in the attenuation of CYP2B1, GST, and catalase proteins and enzyme activities. PMID- 12653019 TI - Percutaneous absorption of topical N,N-diethyl-m-toluamide (DEET): effects of exposure variables and coadministered toxicants. AB - Exposure to N,N-diethyl-m-toluamide (DEET) commonly occurs in the general population and has been implicated as a contributory factor to the Gulf War Illness. The focus of the present studies was to determine the effect of coexposure factors, potentially encountered in a military environment, that could modulate transdermal flux of topically applied DEET. Factors investigated were vehicle, dose, coexposure to permethrin, low-level sulfur mustard, occlusion, and simultaneous systemic exposure to pyridostigmine bromide and the nerve agent stimulant diisopropylfluorophosphate (DFP). Studies were conducted using the isolated perfused porcine skin flap (IPPSF), with a few mechanistically oriented studies conducted using in vitro porcine skin and silastic membrane diffusion cells. DEET was quantitated using high-performance liquid chromatography. The vehicle-control transdermal DEET flux in the IPPSF was approximately 2 micrograms/cm2/h for both 7.5 and 75% DEET concentrations, a value similar to that reported in humans. DEET absorption was enhanced by coinfusion of pyridostigmine bromide and DFP, by the presence of sulfur mustard, or by dosing under complete occlusion. The greatest increase in baseline flux was fivefold. In vitro diffusion cell studies indicated that silastic membranes had two orders of magnitude greater permeability than porcine skin, and showed vehicle effects on flux that were not detected in the IPPSF. These results suggest that coexposure to a number of chemicals that potentially could be encountered in a military environment may modulate the percutaneous absorption of topically applied DEET beyond that seen for normal vehicles at typically applied concentrations. PMID- 12653021 TI - Effects of trimetazidine on acetic acid-induced colitis in female Swiss rats. AB - Induction of colitis by acetic acid (AA) in the rat is widely used experimental model of inflammatory bowel disease (IBD) and ulcerations. AA as an irritant induces colitis involving infiltration of colonic mucosa with neutrophils and increased production of inflammatory mediators, such as hydrogen peroxide (H2O2), nitric oxide (NO), myeloperoxidase activity (MPO), and tumor necrosis factor (TNF alpha). Trimetazidine (TMZ), an antianginal compound, was administered to investigate if its cytoprotective features in cardiac tissue are also effective in AA colitis where ischemic injury contributes to colitis. Administration of TMZ intraperitoneally improved the macroscopic and microscopic score alterations produced by AA. AA administration significantly elevated colonic MPO activity; however, treatment with TMZ significantly lowered this enzyme activity compared to AA. AA administration significantly enhanced superoxide dismutase (SOD) activities, except for AA + TMZ given rectally. TMZ treatment significantly lowered nitrate levels, but AA increased these levels. AA administration markedly lowered TNF-alpha levels, but TMZ treatment elevated these levels to control. These findings indicate that overproduction of NO may be involved in the immunosuppression observed during acute AA-induced rat colitis. In conclusion, TMZ treatment was more effective via the intraperitoneal than rectal route, and may be beneficial in therapy of colitis. PMID- 12653022 TI - Lead in young herring gulls: paradoxical effects of exercise on tissue concentrations. AB - Exposure to lead prenatally and early in life affects physiological, behavioral, and intellectual development in humans and other animals. The movement and storage of lead within tissues and organs and its elimination from the body influence the amounts reaching sensitive target organs such as the developing brain. In this study, young herring gulls, Larus argentatus, were used to examine the effect of mild exercise on the deposition of lead in bone, to clarify possible differences between free-ranging birds in nature and confined birds in the laboratory. Forty 2-day-old chicks were randomly assigned to one of four groups: lead treatment with and without exercise, and controls with and without exercise. Chicks in the lead treatment group received a single intraperitoneal injection of lead acetate solution (100 mg/kg); weight-matched control chicks were injected with an equal volume of isotonic saline at the same age. Chicks in the exercise groups were taken outside and encouraged to move about for 2.5 h/d; others were confined in standard cages. Chicks were sacrificed at 45 d of age and lead was analyzed by graphite furnace atomic absorption. The interaction of exercise and lead dose significantly influenced the amount of lead in the three bones examined (rib, humerus, tibiotarsus), but not in the other tissues. For the lead-treated birds, mean bone lead levels were lower in the exercised versus nonexercised birds, while the reverse was true for control chicks. The data suggest that lead storage can be partly ameliorated by exercise, that young chicks would be the most affected by lead, and that older chicks that move and have larger territories in which to move may suffer lesser effects than those in dense territories. PMID- 12653023 TI - Consistency among methods of assessing concerns about the Los Alamos National Laboratory. AB - In making environmental remediation and restoration decisions, risk assessors and managers need to take into account the environmental concerns of people living around an industrial facility, as well as those residing in the general region. Yet such information is usually anecdotal rather than quantitative, and rarely compares perceptions among different alternatives methods. The concerns of individuals living in Santa Fe, NM, near the Department of Energy's Los Alamos National Laboratory (LANL) were characterized. On an open-ended question, the public's greatest concerns were for contamination (35%), ecological health (16%) and human health (14%). When asked to rate their level of concern from a list of possible concerns, people rated accidents/spills and storage of additional nuclear material the highest, and changes in property values the lowest. Unexpectedly, ethnicity, education, and income did not explain variations in ratings for most concerns about LANL. There was generally agreement between the concerns expressed on the ratings and on the open-ended question, although on the latter individuals expressed concern for larger issues, rather than specific issues. Preferences for future land use reflected their concerns for maintaining a safety buffer of an ecosystem around the site. PMID- 12653024 TI - [Why not include an anti-pneumococcal vaccination in the anti-influenza program?]. PMID- 12653025 TI - [Factors predicting the disappearance of virus B replication markers following treatment with lamivudine]. PMID- 12653026 TI - [Socioeconomic determinism of obesity in the Seine-Saint-Denis area]. AB - OBJECTIVE: Obesity is a complex multi-factorial disease. The role of socioeconomic factors is known but few studies have attempted to analyse separately the impact of the various participating factors: oncome, level of education, cultural and social status. METHOD: These factors were analysed in 26,278 persons aging from 16 to 59 years, living in the district and having benefited from a medical check-up in the Seine-Saint-Denis health and social prevention centre, district particularly concerned by socio-economic insecurity. Among these persons, a representative sample of 1804 filled-in an additional questionnaire including questions on their income, level of education, marital status and the area or country of origin. RESULTS: The prevalence of obesity (body mass index or BMI > or = 30 kg/m2) and overweight (MCI = 25 to 29.9 kg/m2) was respectively of 17.6 and 32.7%. Using univariate analysis, the prevalence of obesity was significantly associated with age, gender (higher in women), settled way of life, socio-professional category, low education, marital status and origin (higher in persons from Africa and North Africa). Using a logistic regression model, the risk of obesity was increased 1.45-fold in persons earning less than 838,47 euros and 1.67-fold in persons with low education. Moreover, it was 2.28-fold greater in the un-working population, 1.62-fold in the redundant and 1.5-fold in the working class, compared with the executive-freelance population. CONCLUSION: The risk of obesity is therefore independently related to cultural, economic and social parameters. PMID- 12653027 TI - [The individual reception project (IRP) for anaphylactic emergencies. The situation in France and French overseas territories in 2002]. AB - INTRODUCTION: The management in schools of children with life-threatening or chronic diseases has led to the development of Individual Reception Projects (IRP) since 1993, notified in the state education ministry's circular letter. The aim of this study was to assess the status of such IRPs in France and the French Overseas Territories in the management of food allergy risks. METHOD: The regional departments of educational promotion were contacted and the survey was run between May and June, 2002. The items of the questionnaire were: the number of IRPs for anaphylactic emergencies, details on the allergies themselves, an overview of the eventual existence of dialogue structures among school physicians, heads of schools, allergists, treatment prescribers and emergency protocols. RESULTS: The total survey revealed 7482 IRP for anaphylactic reactions to food (14% of the total IRPs). The frequency of IRPs varied greatly depending on the regions. The Paris area represented 36.3% of the total. The Lorraine area: 5.6%, the Haute-Garonne: 5%, and the Bouches-du-Rhone: 3.3%. The IRPs represented 0.002 (Ardeche) to 0.33% (Alpes-de-Haute-Provence) of the school attending population. The mean level was of 0.065%; 71.6% of the IRPs concerned the 1st grade (1 IRP for 1091 children). In decreasing frequency, the allergens concerned were: peanuts, eggs, dried nuts, fish, dried peas and milk. Dialogue structures were established in 38 regions. DISCUSSION: The IRPs have increased 4-fold since the circular letter in 1999. The enhanced prevalence of numerous (new) allergies to dried nuts and leguminous plants has been emphasized, and peanut allergies have been confirmed. The complexity of managing the IRPs and need for information and training of state education and municipal staff requires the further development of dialogue structures, which are still clearly insufficient. PMID- 12653028 TI - [Uterine and ovarian metastases 28 years after the discovery of breast cancer]. AB - INTRODUCTION: Breast cancer seldom leads to uterine metastases, but more frequently spreads to the ovaries. OBSERVATION: A 72 year-old woman treated 28 years earlier for a breast cancer (conservative treatment and ovarian castration through radiotherapy) presented with metrorrhagia. The hysteroscopy and uterine curettage were normal. Three months later she presented with bone pains and pain along the course of the femoral nerve. The clinical examination revealed the existence of left axillary and sub-clavicle adenopathies. A biopsy of the lymph nodes showed a lobular adenocarcinoma with positive hormonal receptors. The clinical and biological state of the patient improved under treatment with tamoxifen and aredia. Eight months later the patient complained of pelvic pains. The ultrasound examination showed a pelvic tumour situated at the fundus of an enlarged uterus, as well as peritoneal effusion. The patient underwent a total hysterectomy and multiple biopsies that revealed diffuse metastases of a lobular adenocarcinoma spreading to the neck and the body of the womb, the mesosalpinx, the left ovary and the epiploon. The patient died less than a year after the first symptoms had appeared. DISCUSSION: Uterine metastases of extrapelvic origin are rare. Breast cancer is the first cause of this dissemination. It is generally invasive, lobular, and frequently spreads to the ovaries. In the case reported here, what is remarkable is the exceptionally long period between the discovery of a breast cancer and the apparition of metastases. Hence, in order to detect a primary or secondary ovarian or uterine cancer, it is important to emphasize the necessity of systematic pelvic examination in patients treated for breast cancer. PMID- 12653029 TI - [Compliance to treatment and type 2 diabetes: impact on glycemic balance]. PMID- 12653030 TI - [Mondor's thrombophlebitis and antiphospholipid antibodies]. PMID- 12653031 TI - [Efficacy of cabergoline in the treatment of macroprolactinoma]. PMID- 12653032 TI - [Silent maxillary sinusitis]. PMID- 12653033 TI - [Diagnostic strategy when confronted with a moderate and prolonged increase of transaminases]. AB - TWO TYPES OF SITUATIONS: Measurement of transaminase serum activity is a common biological test. Although the etiological scope of acute and severe hyper aminotransferase is codified and limited, that of prolonged and moderate hyper aminotransferase is much broader. IN THE CASE OF PROLONGED AND MODERATE INCREASE IN TRANSAMINASE SERUM ACTIVITY: The discovery of this abnormality during systematic biological controls is a frequent situation, and its management is relatively well standardised. It requires a rigorous diagnostic strategy, which includes the search for consumption of alcohol, overweight, chronic hepatic disease of viral origin and the nature of the medicinal products ingested. FROM AN ETIOLOGICAL POINT OF VIEW: The most frequent causes of moderate and prolonged hyper-aminotransferase are alcohol abuse, overweight, non-insulin-dependent diabetes, dyslipaemia, viral hepatitis and medicinal products. However, less frequent hepatic or extra-hepatic causes must not be neglected. PMID- 12653034 TI - [Aldosterone and its antagonists in heart failure]. AB - THE ROLE OF ALDOSTERONE: Aldosterone is the key hormone in salt-water homeostasis. In heart failure, it participates in the appearance and maintenance of signs of congestion. Predominantly synthesised in the glomerular area of the cortico-adrenal glands, extra adrenal production areas have recently been identified notably in the brain, the heart and the large artery trunks. Aldosterone is activated in the cells by the intracellular mineral corticoid receptor. IN CARDIOVASCULAR-PATHOLOGIES: In chronic heart failure, patients treated with conversion enzyme inhibitor may escape from the renin-angiotensin blockade and this may lead to increased aldosterone plasma levels. This increase can induce not only vascular lesions and myocardial fibrosis but also renal and cerebral lesions. THE EFFECTS OF SPIRONOLACTONE: In patients with NYHA stage III or IV heart failure, addition of spironolactone to the treatment with conversion enzyme inhibitor, diuretic and/or digitalis leads to a reduction in morbidity and mortality, as demonstrated in the RALES study. The mechanisms by which spironolactone has a beneficial effect remain discussed. IN CLINICAL PRACTICE: The prescription of spironolactone is limited by hormonal side effects it provokes. IN THE FUTURE: Eplerenone, a new competitive aldosterone receptor antagonist that appears to be devoid of such side effects and which, at least experimentally may well have the same beneficial effects, is presently under clinical assessment. PMID- 12653035 TI - [Mass screening of gynecological cancers in public health]. PMID- 12653036 TI - [Promoting medical technologies in the national list of health services: 1995 2000]. PMID- 12653037 TI - [Advanced patient simulators in pre-hospital trauma management training--the trainees' perspective]. AB - BACKGROUND: Computerized Human Patient Simulators (HPS's) have been increasingly utilized to improve and assess medical performance during critical scenarios. AIMS: To evaluate the utilization of advanced HPS's for trauma management training in pre-hospital setting. METHODS: Four scenarios representing frequent critical events in the pre-hospital trauma treatment were developed using the METI (Medical Education Technologies Inc, Gainesville, Florida) and SimMan (Laerdal, Stavanger, Norway) HPS's. Reserve and regular army medical teams encountered the scenarios, which were all fully videotaped. The A-V recording of the training sessions was used in the debriefing session conducted by experts in trauma care immediately after the encounters. At the end of the workshop, all participants filled in a personal feedback questionnaire. RESULTS: Sixty-six physicians and 16 emergency medical technicians participated in the study. Seventy-eight members (95%) of this group had participated in a previous Advanced Trauma Life Support course, 63 (77%) during the two years before the study. Only 6 (7.3%) of the participants treat trauma cases daily, and 7 (8.5%) more treat such cases at least once in two weeks. Seventy-two (82.8%) of the participants pointed out that the scenarios represent true trauma cases, and the same number reported that the video-based debriefing was excellent or very good. Sixty-four participants (78%) reported that the simulator training improved their decision making, 54 (65.8%) claimed that it improved their manual skills, and only 47 (57.5%) believed they gained new knowledge. Eighty participants (97.5%) recommended simulator-based training as part of future training and performance assessment. CONCLUSIONS: According to this preliminary subjective study, simulator-based training may be used as a tool in improving the competency of medical teams in treating trauma victims in pre-hospital settings. PMID- 12653038 TI - [Left paraduodenal hernia: a report of a case and a review of clinical and diagnostic CT findings]. AB - Internal hernias are an uncommon cause of small bowel obstruction. Paraduodenal hernias have been considered until recently the most common sub-type. Due to non specific and intermittent signs and symptoms the diagnosis of these hernias is notoriously difficult. We report a case of a paraduodenal hernia diagnosed correctly with abdominal computed tomography that was confirmed at surgery and review the clinical and imaging findings of these hernias. PMID- 12653039 TI - [Causes of blindness in the year 2000 in Israel]. AB - This study of the causes of blindness in Israel in the year 2000 is based on the National Registry of the Blind in Israel. Patients are considered blind if they have a visual acuity of 1/60 or worse, in their better eye, or visual field loss of > 20 degree, in their better eye. At the end of 1998, 15,937 were registered as blind, 2.7/1000 of the total population and in the year 2000 2.9/1000. The leading causes of blindness in Israel in 1998 were glaucoma, macular degeneration and diabetes mellitus. In the year 2000 the leading causes of blindness are: macular degeneration, glaucoma and diabetes mellitus. Macular degeneration is found to be more prevalent in the year 2000 because of high life expectancy of the total population. Glaucoma is found to be less prevalent then in 1998. This can be attributed to early diagnosis and better treatment of the disease. Early detection of glaucoma and good response to treatment is essential for prevention of blindness in such cases. PMID- 12653040 TI - [Levels of INR in patients admitted to the department of internal medicine]. AB - We reviewed the records of 282 patients admitted to the Department of Internal Medicine who had been treated at home with warfarin. In 181 patients INR was measured on the day of admission. Less than 20% of the 44 patients treated with warfarin for artificial valves had INR levels within the therapeutic range--3-4. Among the 138 patients treated with warfarin for atrial fibrillation only 31.9 were in the therapeutic range of 2-3. Seven patients in the atrial fibrillation group had very high values of INR--above 7. These patients were older (mean age 82 years) compared to 69.4 years in the rest of the group. Four patients with INR's above 7 had mild hemorrhagic events. Our data show that most patients treated with warfarin were not in the therapeutic range. PMID- 12653041 TI - [Updating the basket of health services]. AB - The process by which services are added to Israel's basic basket of health services constitutes a unique effort to tackle the most difficult of ethical issues. In some years, since 1997, the Israeli government allocated an increment of about 1% to the national health budget for the purpose of expanding the basic basket of health services provided under national health insurance. A public committee has been created for this purpose of deciding which of hundreds of potential entrants, should be added within the above budget constraint. The committee integrates available scientific evidence on effectiveness with social values to come up with a ranking. The process has attracted a great deal of public attention, most notably in the printed media, which usually focuses on the plight of specific individuals whose "desired drug" has not been added to the basket. In addition, the courts have, on occasion, intervened to reverse decisions of the committee in the contexts of suits brought by individuals seeking coverage for a treatment not currently included in the basket. Thus, Israel's brave attempt to cope with the necessity of health care rationing needs to be fine-tuned in terms of the information provided to the public about the process and the balance of scientific, economic, judicial and political inputs into the process. PMID- 12653042 TI - [Barriers to the fair allocation of healthcare resources]. AB - The gap between infinite healthcare needs and finite health resources imposes on practicing physicians a dual commitment to patients' needs and to the fair allocation of health care resources. The ethical principles of a fair allocation of resources are egalitarianism, utilitarianism and a transition from individual to group-based ethics. These principles are ambiguous and occasionally conflicting. Their implementation is further impeded by practice norms, such as private practice by physicians employed by public institutions, incentives for physicians to save, and marketing of clinical interventions of unproven efficacy. Proposed approaches to these barriers to the fair allocation of resources include respect of patient autonomy with full disclosure of the limitations in the services provided by the health plan; adherence to clinical practice guidelines; and an open access for patients to second medical opinion. I believe that the fairness of resource allocation may be further improved by the prohibition of private practice by physicians employed in public health care institutions, and by enhancement of physicians' self-awareness of subconscious discrimination against some patients. PMID- 12653044 TI - [External cephalic version]. AB - Breech presentation is the most common of malpresentations, occurring in 3-4% of all term deliveries. The fetus may adopt breech presentation because of many factors. In the past, the mode of delivery (vaginal or abdominal) was determined by clinical considerations. Current recommendations are for cesarean section in all breech presentations. Successful External Cephalic Version is the only effective way to avoid cesarean section in breech presentation. Considering success rates and related factors, this review article emphasizes the importance of considering ECV to every woman with breech presentation. PMID- 12653043 TI - [The use of cardiac troponins in acute coronary syndromes]. AB - In patients with acute coronary syndromes cardiac troponins are sensitive markers of myocardial damage. The troponin complex comprises subunits C, T and I and is a major component in the process of myocyte contraction and relaxation. The T and I subunits have cardiac isoforms with distinct specific immunologic properties which distinguish them from similar subunits of non-cardiac muscle tissue. The high sensitivity and specificity of cardiac troponins make them the preferred biochemical markers for diagnosing acute myocardial infarction and for the triage of patients admitted with chest pain without ST segment elevation on the E.K.G. There is a correlation between cardiac troponin levels and prognosis in patients with acute coronary syndromes. In addition to their prognostic role, cardiac troponins play a role in selecting patients for contemporary treatments. Thus, their level can identify the patients most likely to benefit from treatments such as low molecular-weight heparin, IIb/IIIa receptor blockers and early angiography and coronary intervention. Recently, the American heart Association, the European Heart Society and the Israel Cardiology Society have published guidelines for the use of cardiac troponins. This review summarizes the current data regarding the use of cardiac troponins in the acute coronary syndromes. PMID- 12653045 TI - [Scales for evaluation of the geriatric patient with hip fracture during the acute and rehabilitation phases]. AB - The elderly population is increasing in the modern world. Societies in general and medical personnel are facing new ethical and medical dilemmas when treating extremely old patients. Many factors are known to cause fractures in elderly patients. Probably, the most important one is osteoporosis, which was shown to be the main pathology in elderly patients who fractured their hips. The increase in fall frequency, orientation to falls and the protective response to trauma were found to play a role in the pathogenesis as well. The rehabilitation of this group of patients is an important factor that helps to return the elderly patient to his or her own environment and to his or her skills prior to the falls. The decision of the potential for rehabilitation and the evaluation of the progress that the elderly does is very difficult and can vary substantially from one examiner to another. Reviewing the literature that exists on the evaluation of the geriatric patient with hip fracture during the acute and rehabilitation phases is our goal. PMID- 12653047 TI - [Adolescent pregnancy in Israel: a methodology for rate estimation and analysis of characteristics and trends]. AB - PROBLEM: Teenage pregnancies are frequently unplanned, the young pregnant woman being often unmarried. These pregnancies occasionally end in induced abortion. Teenage pregnancy, especially when unintended, is correlated with negative effects for the child and the mother. Despite the need to reduce the magnitude of this phenomenon, there is no reliable database on teenage pregnancy. Such a database could help define populations at risk, and could help in designing prevention programs. Birth rates, fetal deaths, and induced abortion are reported, but there is no uniformity or merging of the data. OBJECTIVES: The aims of the paper are to present a methodology for the estimation of pregnancy rates and to use it for the analysis of characteristics and trends in adolescent pregnancies in Israel. METHODS: The pregnancy rate was calculated as the sum of three outcomes: live birth, induced abortion, and fetal loss, at all stages of pregnancy. RESULTS: Based on these computations, the teenage pregnancy rate in Israel was estimated as 32 per 1,000 adolescent girls. This rate was three times higher among Moslems as compared to Jews. From 1988 through 1998, the rate decreased by 12%. The pregnancy rate in Israel is similar to the rate of most Western-European countries, and is three times lower than the US rate. CONCLUSION: There is a need to improve the means for collecting the data on which the computations are based, to build an algorithm specifically adapted to Israel, and to publish detailed pregnancy rates on a regular basis. PMID- 12653046 TI - [A practical noninvasive diagnostic approach to acute venous thromboembolism in the emergency department]. AB - The diagnosis of venous thromboembolism (VTE) (deep vein thrombosis and pulmonary embolism) poses a challenge for physicians in emergency departments (EDs). Clinical findings and non-specific laboratory tests are usually insufficient to rule out VTE. Due to high costs, higher complication rates, and other reasons, the "gold standard" tests for VTE (pulmonary angiography or contrast venography) are usually unavailable and are seldom used by most physicians in the EDs. The end result is an extensive use of the non-invasive Imaging modalities and yet under-diagnosis of VTE. The purpose of this review is to describe the advanced non-invasive methods currently available for the diagnosis of VTE and to define the role of the rapid d-dimer assay. We also propose a practical diagnostic protocol for VTE that is based on a combination of D-dimer test and judicious use of non-invasive imaging, which may be implemented in all emergency departments. PMID- 12653048 TI - [Steroid induced glaucoma]. AB - Steroid treatment has gained notoriety due to its tendency to induce multiple side effects, including a variety of ocular side effects. Administration of local, regional, inhalation or systemic steroids may induce the development of ocular hypertension, which might even result in subsequent open angle glaucoma. About one in every three people is considered a potential "steroid responder". A significant elevation of intraocular pressure might result in these patients in response to steroid treatment. Included in this group are patients with first degree relatives suffering from open angle glaucoma. Morphologic changes in the trabecular meshwork (which serves as the site of aqueous humor drainage from the eye) are suggested as the proposed mechanism through which steroid treatment results in glaucoma. Steroids are said to induce the expression of a gene that is located on chromosome 1 and is known as TIGR or GLCIA. its product is a protein called myocilin. Ocular hypertension secondary to steroid treatment is usually reversible, when treatment is limited to a period of less than 12 months. The fear of ocular hypertension, which is usually unnoticed by the patient, obligates regular ophthalmologic follow-up examinations, including tonometry, visual fields and optic disc examinations. PMID- 12653049 TI - [Representing patients hospitalized under legal commitment at district psychiatric committees]. AB - The Tel Aviv-Central District Office of the Public Defenders' Office has begun, as a trial project, to represent patients hospitalized under enforced commitments at district psychiatric committee hearings concerning their welfare. This experimental trial was carried out at the Abarbanel Mental Health Center starting 1st January 2000. The results illustrate that the chances of a patient hospitalized under enforced commitment being discharged from hospital are better if he is represented. It is particularly better when cooperation exists between his public defender and his treating psychiatrist. The results emphasize that the inclusion of the defender in the process affects the work program of the committee and its decisions, especially the examination of facts, the hearing of witnesses, the quality of the psychiatric assessment, the examination of the legal aspects and the summing up of the findings. From feedback received, it appears that all parties involved in the process feel that representation on behalf of the patients by the legal defenders results in more just and worthy decisions in every public aspect of the process. The authors recommend that legal representation be extended to all patients who are hospitalized under enforced commitments. PMID- 12653050 TI - [Juvenile polyposis in Mexican children]. AB - BACKGROUND: Juvenile polyposis (JP) is a frequent cause of lower gastrointestinal bleeding. It is present in 3-4% of the population < 21 years of age and represents 90% of all polyps in childhood. The most common complaints are hematoquezia, abdominal pain, and polyp prolapse. METHODS: A descriptive, observational, transversal study of 225 children with histopatologic diagnosis of JP seen at the Department of Gastroenterology and Nutrition of the Instituto Nacional de Pediatria, between January 1985 and December 2000. Variables studied included age of presentation, gender, frequency, clinical manifestations, type, location, and diagnostic and therapeutic methods. RESULTS: Frequency was one polyp per 162 patient seen during the same period (0.61%) and age 2 to 8 years (82%), a total of 235 patients, 120 females and 273 total polyps were found rectum the most common site 82% (224 polyps) followed by sigmoid. Mean distance from anal margin was 6.5 cm. Of all polyps, 82.9% were in the first 10 cm of anal margin. Of all patients, 92.3% had a single polyp and 7.7% had multiple polyps. A baritated study was made in 39 patients (16.6%) with diagnostic efficacy of 74%. No additional baritated studies were needed because of high diagnostic and therapeutic efficacy of the rectosigmoidoscopy. CONCLUSIONS: Rectosigmoidoscopy and colonoscopy remain as the diagnosis and therapeutic method in children with JP. PMID- 12653051 TI - [Helicobacter pylori genotypes and their association with host's immune response]. AB - BACKGROUND DATA: The study of Helicobacter pylori phenotypes and genotypes is mainly focused on two groups of putative bacterial virulence factors: the cag pathogenicity island (PAI), for which CagA is a marker, and the vacuolating cytotoxin VacA. Several studies have shown the clinical relevance of the determination of IgG anti-CagA antibodies. OBJECTIVE: To assess the prevalence of vacA and cagA genotypes of H. pylori and the association with IgG anti-CagA antibodies in symptomatic patients. METHODS: We studied 50 patients (mean age 53 years, range 15-92). PCR amplification of the vacA s and m regions was performed using the primers described (J Biol Chem 1995; 270: 17771). For cagA PCR, primers described by Rugge et al. were used (Cancer 1999; 85: 2506) and determination of IgG anti-CagA antibodies was done according to the method described by Blaser and Perez (Cancer Res 1995; 55: 2111). RESULTS: All 50 patients studied were positive for H. pylori. Of the 50 H. pylori strains, 7 (14%) were isolated from patients with peptic ulcer disease and 43 (86%) from patients with non-ulcer dyspepsia. The most frequent vacA genotype was s2/m2, associated with cagA-H. pylori strains (p < 0.01). Presence of cagA+ H. pylori strains correlated with the presence of IgG antibodies (Kappa = 0.680). Determination of IgG anti-CagA antibodies showed a sensitivity of 77.4%, specificity of 94.7%, positive value of 96% and negative predictive value of 72%. CONCLUSIONS: The most frequent H. pylori genotype found in northeastern Mexico was vacA s2/m2, cagA-. The presence of this genotype correlated with the clinical presentations observed in these patients. In addition, CagA serology showed a great specificity and good sensitivity that allow us to use this assay to assess the prevalence of CagA+ strains in Mexico. PMID- 12653052 TI - [Differences in the stress symptoms rating scale in Spanish between patients with irritable bowel syndrome (IBS) and healthy controls]. AB - Irritable bowel syndrome (IBS) can be explained by the interrelation of cognitive components such as illness behavior, emotional such as depression and anxiety, behavioral such as environmental stressors, and physiologic such as pain modulation alterations. The stress symptom rating (SSR) scale is being used in physiologic studies in IBS, but differences between IBS patients and healthy controls in the basal condition is unknown. AIMS: To determine the basal differences in the SSR-Spanish between IBS patients and controls, and between IBS patients according to bowel-habit predominance. METHODS: Forty consecutive IBS Roma 1 patients [M: 28 (78%), F:8(22.2%), 42.6 +/- 2.7 years] and 36 healthy volunteers paired by gender and age [M: 28 (78%), F:8 (22.2%); 36.7 +/- 2.1 years] were included. IBS were classified as constipation, diarrhea, or alternating. All completed the SSR prior to the medical visit. Mean group values of the different SSR parameters were compared by ANOVA. A p < 0.05 was considered significant. RESULTS: Compared with controls, IBS patients reported higher anxiety (p = 0.005), attention (p = 0.04) fatigue (p = 0.04), and lower arousal (p = 0.003). There were no differences in stress and anger either in IBS patients according to bowel-habit predominance. CONCLUSIONS: Under basal conditions, there are differences in anxiety, weakness, attention, and arousal between IBS patients and controls that correspond to intrinsic characteristics of IBS independent of bowel-habit predominance. Higher anxiety is in agreement with psychological comorbidity, higher attention with hypervigilance, and fatigue and lower arousal with sleep disturbances known to be present in IBS. Absence of differences in stress and anger under basal conditions suggests that SSR-Spanish is a good instrument to evaluate response to stress during physiologic paradigms. PMID- 12653053 TI - [Selection criteria for endoscopic cholangiopancreatography before laparoscopic cholecystectomy]. AB - BACKGROUND: The unexpected detection of stones in common bile duct during laparoscopic cholecystectomy is a concern for laparoscopic surgeons. Immediate laparoscopic removal may not always be feasible due to inadequate operating facilities, surgeons, and assistants. Attempts have been made to identify clinical and biochemical predictor of common bile duct stones, the results of which have not been consistent. OBJECTIVE: The aim of this study was to define patients with risk for common bile duct stones based on history, abdominal ultrasound, and biochemical derangements, as well as to evaluate the role of endoscopic retrograde cholangiopancreatography (ERCP) and endoscopic sphincterotomy (ES) for diagnosis and duct clearance if necessary. PATIENTS AND METHODS: This is a prospective cross sectional analysis of 82 patients (66 females and 16 males) with gallstones on whom were performed clinical examination, laboratory test, and abdominal ultrasound to identify predictors of common bile duct stones. Preoperative ERCP was carried out and patients underwent ES for stone retrieval. RESULTS: Clinical evidence of jaundice or pancreatitis, elevated serum amylase, and ultrasonographic evidence of biliary tree dilation or common bile duct stone were considered risk factors. In 45 patients, common bile duct stones were identified at preoperative ERCP patients underwent ES, and stones were removed with 100% success. All patients were scheduled for laparoscopic cholecystectomy 24 h later. There was no mortality in this series. CONCLUSIONS: Clinical evidence of jaundice or pancreatitis, elevation of serum amylase and dilated common bile duct or presence of common bile duct stones were considered risk factors for choledocholithiasis. Use of such a model rather than individual criteria would improve selection of patients for preoperative ERCP, optimizing its role in the laparoscopic era. PMID- 12653055 TI - [Surgical treatment of unspecific chronic ulcerative colitis. A 5-year experience]. AB - BACKGROUND: Treatment of ulcerative colitis depends on clinical stage of disease and have precise surgical indications. The aim of this work was to review surgical indications and present data related to our experience. MATERIAL AND METHODS: Retrospective, descriptive, and transversal study. We reviewed records of all patients who underwent a surgical procedure for ulcerative colitis at our service from March 1996 to March 2001. RESULTS: Twenty two patients, 13 males and nine females, ages range 21 to 72 years. Main indication for surgery was no response to medical treatment (50%). Surgical procedures was subtotal colectomy with ileostomy in one patient, intersphincteric proctocolectomy with ileostomy in four intersphincteric proctectomy with ileostomy in three ileorectalanastomosis in three proctocolectomy with ileoanal "J" pouch in six and proctectomy with ileoanal J pouch in five. We had the following complications: oral candidiasis; phlebitis; eventration; pouchitis, and anal fissure in one patient, respectively, pouch-skin fistula in two patients (9%), and retrograde ejaculation in one of these. Follow-up was for 5 years. CONCLUSIONS: Surgical indications for ulcerative colitis are precise. We recommend early surgical evaluation. Ileoanal pouch is now considered the gold standard for surgical procedure because it eliminates disease, neoplasm development, and permanent extra-colonic manifestations, and restores continence. PMID- 12653054 TI - [Ischemic colitis in patients submitted to aortic replacement surgery. Risk factors]. AB - Ischemic colitis (IC) is an important clinical problem, and may present after aortic surgical procedures. The aim of this work was to establish risk factors for IC presentation in aortic surgical replacement patients. MATERIAL AND METHODS: A retrospective study of patients with aortic surgical replacement in a 3-year period was carried out. Patients were divided into two groups: patients without IC and patients with IC, the later group subdivided into patients with gangrenous ischemic colitis and without gangrenous ischemic colitis. Multiple logistic regressions was used to obtain the variables for possible risk factor for IC. RESULTS: We included 101 patients in the study; ischemic colitis was present in 16.8% of all cases, with 47.1% of gangrenous type. Metabolic acidosis was the most frequent alteration. Diagnosis was made by endoscopy in 94.1%. Mortality in IC group was 18.2% with an increase in the gangrenous group to 62.5%. Identified risk factors were disrupted aneurysm, previous colonic surgery, emergency surgery, and hemodynamic instability. CONCLUSIONS: Ischemic colitis is most frequent in emergency surgery for disrupted aneurysm in the hemodynamically unstable patient with retroperitoneal hematoma. We must entertain a high suspicion index for IC in all patients with aortic surgical procedures for early detection and adequate treatment. PMID- 12653056 TI - [Esophageal closed transection for surgical treatment of portal hypertension. A 10-year experience]. AB - BACKGROUND: Extensive esophagogastric devascularization (Sugiura-Futagawa operation and complete portoazygos disconnection) are excellent alternate choices for low-risk patients (Child-Pugh A-B) in whom a selective shunt is not feasible, obtaining a low postoperative rebleeding rate and a good quality of life. Esophageal transection is an important step in these procedures because submucosal varices are obliterated, achieving complete portoazygos disconnection. Results during one decade with closed variant of esophageal transection are reported. METHOD: Between 1990 and 2000, 78 patients were operated on (69 Child Pugh A and nine Child Pugh B) in two operative stages with Sugiura-Futagawa operation and complete portoazygos disconnection. Files of these patients were reviewed and recurrence of hemorrhage was evaluated as well as fistulization and dehiscence. RESULTS: Among 78 modified transections, only one case of dehiscence was observed (1.2%) (very early in our experience) with concomitant mortality. Rebleeding was observed in 11% of cases, with two cases of stenosis that required dilatation. During the last 7 years (57 patients), no complication has been observed. CONCLUSIONS: Modified transection has a very low complication rate with long-term results comparable to those of classic transection. PMID- 12653057 TI - [Boerhaave's Syndrome. Case report and literature review]. AB - OBJECTIVE: Report of a case of spontaneous perforation of esophagus in a 70-year old patient. BACKGROUND: Boerhaave syndrome or spontaneous esophageal perforation is a life-threatening condition that demands early diagnosis and urgent management. Without surgical intervention, spontaneous perforation of esophagus is virtually incompatible with life. The classic history is of a patient who vomits and experiences epigastric or substernal pain. Diagnosis is often delayed because esophageal perforations can masquerade as many other clinical disorders. METHODS: Report of a case of a patient with Boerhaave syndrome referred to our institution. CONCLUSION: Boerhaave syndrome is suspected on a clinical basis and confirmed with radiologic studies. As gastric content passes to mediastinum and usually pleural space, a delay in diagnosis raises morbidity and mortality significantly. This pathology is best treated with definitive repair and mediastinal and/or pleural drainage procedures. PMID- 12653058 TI - [Splenic hydatid cyst. Case report of a pregnant woman]. AB - Hydatid disease is caused by larvae of the platyhelminth parasite called Echinococcus; it generally lodges in liver and lung, and less commonly in spleen. In Mexico, hydatid disease has a very low frequency; only few cases of lung disease have been reported. In this paper, case of a 7 weeks pregnant female patient with a simple cyst in spleen identified by ultrasound and tomography is reported. The patient underwent splenectomy. Diagnosis of hydatid cyst was confirmed by histopathology. This case suggests that hydatid disease should be considered as differential diagnosis in every patient with a cystic mass of the spleen. PMID- 12653059 TI - [Perianal leukemic infiltration. Case report]. AB - BACKGROUND: Leukemic infiltration of perianal skin (leukemia cutis) is quite uncommon, and may consist of no diffuse infiltration, erythema, ulceration, fistula, and abscess. Diagnosis may be established by biopsy and adequate hematologic studies. Treatment must be conservative in patients with poorly controlled disease (sitz baths, antibiotics, radiotherapy), and surgical in patients with risk of fulminant sepsis and death. OBJECTIVE: To report a case of leukemic perianal infiltration with atypical ulcerated lesion. CASE REPORT: Female patient, 21 years of age with diagnosis of AML M7, began with perianal pain posterior to evacuation associated with bleeding; at perianal examination we found anterior atypical ulcerated lesion. We carried out a biopsy under suspicion of leukemic infiltration corroborated by histopathology. The patient received conservative treatment. CONCLUSIONS: We must suspect leukemic infiltration in patients with anal symptoms and effect close surveillance. Prognosis depends on control of hematologic disease. PMID- 12653061 TI - [Biliary-colonic fistulas. Analysis of 2 cases and literature review]. AB - BACKGROUND: Bilioenteric fistulas are entities whose etiology can be diverse. OBJECTIVE: The aim of this paper was to present two cases of biliary-colonic fistulas with different presentations and to review existing literature regarding etiology, diagnosis, and management of these patients. METHODS: Two cases of biliary-colonic fistulas were reviewed. RESULTS: Case 1. A 48-year-old woman presented with lower gastrointestinal bleeding secondary to cholecystocolonic fistula in context of cholelithiasis. Case 2. A 60-year-old woman with multiple hepatic abscess associated with biliary-colonic fistula secondary to adenocarcinoma of gallbladder. CONCLUSIONS: Biliary-colonic fistulas are recognized complications of cholelithiasis. Association with neoplasia is less frequent. Preoperative diagnosis is difficult and management must be surgical. PMID- 12653060 TI - [Para-stomal hernias. Report of 3 cases]. AB - Three cases of stomal hernia are presented in this article; two females and one in male patient, the first two cases with colostomy and the remaining patient with ileostomy. One case was treated by replacing stoma and subsequent hernioplasty. The additional two were treated with Leslie technique. To date, there have been no recurrences. Patients were followed for a median of 2 years. Although this complication is infrequent we recommend careful planning prior to performing a stoma, as well as determining the best anatomic site and use good surgical technique. PMID- 12653062 TI - [Pneumoperitoneum]. PMID- 12653063 TI - [Diaphragmatic hernia]. PMID- 12653064 TI - [Massive childhood trichocephaliasis]. PMID- 12653065 TI - [Familial enterobiasis (oxyuriasis)]. PMID- 12653066 TI - [Gastric band complication in morbid obesity]. PMID- 12653067 TI - [Gastroesophageal reflux disease. Gastroenterology Mexican association consensus. Mexican group for the GERD study]. PMID- 12653068 TI - [Application of chromoendoscopy to evaluate the severity and interobserver variation in chronic non-specific ulcerative colitis]. AB - BACKGROUND: Ulcerative colitis (UC) is a disease of unknown etiology characterized by chronic and superficial inflammation of colorectum. Chromoendoscopy has been informed as a method to improve evaluation of extension of UC. No reports on this issue have been published in Mexico to date. OBJECTIVE: To investigate usefulness of chromoendoscopy in determination of extension and severity of UC, as well as inter-observer variability. PATIENTS AND METHODS: Twenty five patients with diagnosis of UC were selected from March 2001 to January 2002 to enter this prospective study. Baron scale was used to evaluate UC endoscopic severity and was followed by randomization of patients to receive methylene blue 0.2% (14 cases) or indigo carmine 0.1% (11 cases) for chromoendoscopic assessment. Each case was presented for evaluation to five endoscopists with > 5 years experience in colonoscopy. Histopathologic report was considered gold standard. STATISTICAL ANALYSIS: Weighed Kappa coefficient for endoscopic-pathologic agreement among evaluators was used. RESULTS AND DISCUSSION: There was fair agreement among endoscopic and pathologic diagnoses of each observer with the two tinctures, particularly when using indigo carmine. Chromoendoscopy increased detection of areas affected by UC with minimal or silent activity. CONCLUSIONS: Chromoendoscopy with indigo carmine or methylene blue may increase endoscopic-pathologic agreement for assessment of severity of UC, thus being a helpful complementary technique in these patients. PMID- 12653069 TI - [Hirschsprung in children over 6 years of age]. AB - INTRODUCTION: Hirschsprung's disease or colonic agangliosis is a rare condition in schoolchildren and adolescents. It is common in infants and chronic constipation is the usual symptom. MATERIALS, METHODS, AND RESULTS: In a period of 5 years, 11-years-old patients were diagnosed and treated for Hirschsprung's disease. We studied six male patients; mean age was 8.6 years. All patients had classic disease and nine, severe chronic constipation, and two were admitted with encopresis. One had 10 kg of feces in 40 cm of colon. Diagnosis was established with barium enema and tissue biopsy. All patients had colostomy. In eight patients, Duhamel procedure was performed with good results. CONCLUSIONS: This disease is a rare condition in older patients and Duhamel procedure is a good surgical option. PMID- 12653070 TI - [Iatrogenic injury of the bile duct. Experience with repair in 180 patients]. AB - OBJECTIVE: Biliary duct lesions have a prevalence of 0.3-0.6% This prevalence is independent of the learning curve: The present paper evaluated survival and quality of life of patients following operative repair. METHODS: In a 12-year period, 180 patients underwent bile duct reconstruction. Of these patients (61 males and 129 females, mean age 39 years), 52% sustained injury during open operation and 42% during laparoscopic procedure. Quality of life was evaluated in the postoperative period. RESULTS: All 180 patients were treated surgically by means of Roux en Y hepaticojejunostomy. Transhepatic stents were used in 142 patients. Eight patients had independent left and right duct anastomosis and in 51 cases, partial resection of segment IV of liver to improve exposure of hilus was carried out. Mortality was 1.7%, due to multiorganic failure. After removal of stent, radiologic manipulation was required in 16% of cases to remove debris and stones or to dilate anastomosis. A total of 83% of patients were completely rehabilitated both clinically and biochemically. CONCLUSIONS: Bile duct injury has a good prognosis in specialized tertiary-care centers. Roux en Y hepatoyeyunostomy is procedure of choice with trans-hepatic stent when needed. Full rehabilitation can be achieved in 80% of patients. PMID- 12653071 TI - [Epidemiologic factors and results of treatment in peripheral and peri-hilar cholangiocarcinoma]. AB - OBJECTIVE: Cholangiocarcinoma is a rare malignant tumor that may occur anywhere along intra or extrahepatic biliary tree. Prognosis remains poor with overall 5 year survival rate of 5%. Experience in management of this lesion in Mexico is scarce. Our objective was to report on our experience at the Instituto Nacional de Cancerologia in Mexico City with the management of this lesion. PATIENTS AND METHODS: A retrospective review of clinical files of patients diagnosed and treated in a period of 10 years was performed. Epidemiologic data, forms of diagnosis, results of management, and survival were analyzed. RESULTS: From June 1992 to June 2002, 36 patients were diagnosed and treated; 25 patients had perihilar tumor (Klatskin tumor) and 11 had peripheral cholangiocarcinoma. Twenty seven were females (75%) and nine males (25%). Abdominal pain was the main symptom (72%) followed by jaundice (65%) and weight lost (59%). Computed tomography was the most used imaging study (86%); surgery was main form of management (72%). No survivors were observed in patients nor receiving treatment after a follow-up of 3 years. For those resected, an 18%, 5-year survival was observed. CONCLUSIONS: This lesion was more frequent in females between fifth and seventh decades of life and no relation with ulcerative colitis or primary sclerosing cholangitis was observed in our study. Surgery is still the best form of management but prognosis remains poor. PMID- 12653072 TI - [Endoscopic diagnosis and treatment of non-traumatic bilhemia. A case report]. AB - Biliovascular fistulas are abnormal communications with two types of clinical manifestations depending on type of flow in fistulous tract: 1) hemorrhage into biliary tract known as hemobilia, or 2) bile into bloodstream, known as bilhemia. Historically, this complication has been treated with surgery; however, technological progress at present allows treatment with intervention techniques without surgery being mandatory. In 1975, Clemens and Wittrin introduced the term bilhemia, a rare complication of hepatic damage producing excessively high levels of serum bilirubin and moderate rise of hepatic enzymes secondary to post traumatic intrahepatic biliovenous fistula. Although this pathology is rare, it is considered dangerous; of 50 patients reported in the literature, 25 died due to this problem. The main purpose of treatment is to release tract obstruction by endoscopic sphincterotomy of Vater's papilla or, if the process is localized in proximal areas of biliary tract, through percutaneous biliary drainage or preferably nasobiliary drainage with continuous suction. This procedure can at least produce temporary relief and occasionally fistula closure. A longer effect can be achieved with biliary stent placement. We describe what is, to our knowledge, the first case of diagnosis and successful treatment of non-traumatic bilhemia with endoscopic sphincterotomy and nasobiliary catheter placement. PMID- 12653073 TI - [Solitary jejunal diverticulum with heterotopic gastric mucosa: diagnosis and management. A case report]. AB - INTRODUCTION: Diverticular disease of small bowel is an uncommon entity and may be the cause of vague abdominal symptoms generally an incidental finding that occasionally require surgical therapy for resolution. METHOD: We report a case of a patient with complicated solitary jejunal diverticulum with perforation and non Meckelean heterotopic gastric tissue that required resection, and include a review of the literature of this association. CONCLUSION: Surgical treatment of diverticular disease of the small bowel is limited to symptomatic patients with non-response to medical therapy or who have some acute complications of same. PMID- 12653074 TI - [Extracolonic manifestation as first sign of colorectal carcinoma. Report of 2 cases]. AB - INTRODUCTION: In Mexico, colorectal cancer is the second most common malignant tumor of digestive tract; incidence is proportional to patient age. At time of diagnosis, 20% of patients can be found with metastases, liver and lung the organs affected most frequently. OBJECTIVE: To present two young patients, whose primary complaints were atypical extracolonic symptoms of colorectal cancer. CASE DESCRIPTION: CASE 1: A 44-year-old female who presented with left pleural effusion, with growth in culture positive for enterobacterias. Barium enema showed tumor in splenic flexure. Surgical resection was performed including, left kidney, spleen, tail of pancreas, and fourth portion of duodenum. Colo-pleural fistula was repaired with primary closure. The pathology report noted locally advanced adenocarcinoma of colon (T4 N0 M0). CASE 2: A 22-year-old female, who presented with a 30 day history of neurologic complaints. Computer tomography of head and MRI of brain identified a mass at level of cerebellum; specimen biopsy report was consistent with metastasis adenocarcinoma. Colonoscopy found a tumor located 12 cm from anal verge. The patient underwent sigmoid resection. Pathology confirmed the presence of moderately differentiated adenocarcinoma and 14/22 positive nodes (T3, N2, M1). CONCLUSION: The most common sites of metastases secondary to colorectal cancer are liver and lung, up to 30% at diagnosis, and usually are asymptomatic. We reported two patients whose first evidence of disease were atypical symptoms related to unusual sites of colorectal cancer extension. PMID- 12653076 TI - [Diabetes mellitus in acute pancreatitis]. AB - Exocrine and endocrine components of pancreas are interrelated anatomically and functionally. Exocrine pancreatic dysfunction often accompanies endocrine pancreatic impairment and vice versa. Diabetes mellitus resulting from alterations of exocrine pancreas, such as acute or chronic pancreatitis, is known as pancreatic diabetes. Hyperglycemia during acute pancreatitis (AP) can be due to abnormalities in insulin secretion, increase in counterregulatory hormones release, or decrease in glucose utilization by peripheral tissues. Causal association is suggested between diabetic ketoacidosis and AP and is attributed to alternation in metabolism of triglycerides. High blood glucose levels are associated with severe AP and constitute factor of worst prognosis. Some patients are discharged with diabetes after AP episode, while others develop diabetes during first year of follow-up. Origin and frequency of glycemic abnormalities associated with AP have not been settled yet accurately. Also, predictive factors for diabetes development and persistence after AP have not been recognized to date. PMID- 12653075 TI - [Colonic stenosis secondary to non-steroid anti-inflammatory agents as a cause of anemia and chronic diarrhea]. AB - Gastroduodenal mucosal injury is a widely recognized side effect of non-steroidal antiinflammatory agents (NSAID). Distal small bowel and colon are additional organs of the gastrointestinal tract exposed to deleterious effects of these drugs. Inflammation and ulceration have been described as pathologic damage associated with NSAID. Strictures of colon induced by NSAID are a new entity characterized by diaphragm-like strictures. Most patients present with anemia, obstructive symptoms, diarrhea, or weight loss. Endoscopic dilation, surgical resection, symptomatic treatment, and interruption of NSAID ingestion are treatment of choice. Only 23 cases of NSAID-related, colonic, diaphragm-like strictures have been reported. Here we describe a case of concentric colonic stricture related to naproxen and clinical features of this entity are discussed. PMID- 12653077 TI - The future of the American Group Psychotherapy Association: whither we goest? AB - The author describes his perspective about what the future holds for the American Group Psychotherapy Association (AGPA). Utilizing Nitsun's (1996) construct of the "anti-group" as an organizing metaphor, he delineates the destructive forces both within and outside the organization that are currently affecting it, and then describes how they can be transformed into instruments for positive development. He suggests that the AGPA can and must continue to provide a "holding environment" (Winnicott, 1960) for professionals interested in working with people in groups. In addition, he asserts that the organization will build upon its remarkable response to the September 11, 2001 debacle in the United States to expand its services to those who have been traumatized. Beyond this, he underscores the importance of the AGPA consolidating and expanding its various educational offerings into a coherent package that will meet the needs of any and all who are interested in learning about how to work effectively with groups. In these and other ways, he contends that the AGPA will thereby confirm Nitsun's assertion that anti-group forces can be transformed, and that organizations that confront these forces constructively can not only survive but thrive. PMID- 12653078 TI - Primary process and peer consultation: an experiential model to work through countertransference. AB - Various models exist for peer supervision and consultation of group therapy. This article documents the authors' experience using an experiential group consultation of group therapy model that relies on primary process to overcome countertransference dilemmas. A review of group therapy supervision and consultation models is followed by vignettes from the authors' experience. Discussion of the vignettes highlight critical issues in group consultation and expound upon the strengths and challenges of using an experiential model. PMID- 12653080 TI - Self-disclosure with all-male groups. AB - Given the relational deficits men bring to a group, therapist self-disclosure is particularly pertinent with all-male groups. The use of this intervention reflects the author's long experience with such groups. The author writes from an interpersonal, collaborative model of group therapy, and discusses types of self disclosure and the guiding principles in their use. Drawing primarily from his own practice, the author presents and discusses clinical vignettes that illustrate both helpful and nonhelpful uses of self-disclosure. PMID- 12653079 TI - Hamlet at the millennium: assisting today's teens through the group psychotherapy process. AB - This article reviews Shakespeare's Hamlet and its universal appeal 400 years after it was written. Focusing on the soliloquy "to be or not to be," the author notes that the play examines the fundamental philosophical question of whether to continue with life, or not. The article then discusses difficulties experienced by adolescents at admission to an inpatient psychiatric program by referring to them as modern day Hamlets. An examination of the theme of ambivalence as it complicates the assessment and treatment process is undertaken. A checklist of therapist responses to ambivalence in the conducting of inpatient group psychotherapy in an adolescent psychiatric hospital unit is offered. PMID- 12653081 TI - Bonding in group: the therapist's contribution. AB - Bonding is an ongoing aspect of intersubjective experience, a type of mental relationship to oneself and others. As a behavioral mode, bonding utilizes verbal and nonverbal communication primarily to establish a basic feeling of connection between individuals. Bonding is the floor, a cognitive-affective state that precedes and prepares the way for the complex processes underlying identification. The concept relates to Bion's early theory of basic assumptions, and takes on new and different meanings according to his later ideas regarding social communication. Bonding is also a therapeutic posture or technique, an element in the establishment of group cohesion, the positive transference, and therapeutic and group alliance. Case examples illustrate how the therapist and other group members bring a complex of bonding needs and anxieties that must be negotiated to establish and maintain bonding, which is a necessary condition for successful therapeutic work. PMID- 12653082 TI - [New therapeutic approaches to age dependent macular degeneration]. PMID- 12653083 TI - Vascular clips as a landmark for sites of anastomosis. AB - Vascular clips can be used in bypasses at risk of future occlusion, to assist with catheter placement for radiological intervention. We explain how vascular clips can be placed as a landmark for the sites of anastomosis. After completion of the vascular anastomosis, two or three bullets of medium-sized vascular clips are clipped to the tips of the vascular suture. The sites of the vascular anastomosis are checked by plain X-rays. Finding the inlet of the anastomosis in an occluded graft can be difficult, and the marker proved helpful for introducing the guide-wire through the inlet of the site of anastomosis. The vascular clips around the popliteal anastomosis enabled us to clearly differentiate above-knee from below-knee femoropopliteal bypasses using plain X-ray postoperatively. This is a simple yet effective method of identifying the site of vascular anastomosis. Thus, we recommend using vascular clips as a landmark to aid in postoperative intervention by allowing clear identification of the level of anastomosis. PMID- 12653084 TI - Doctors and managers. "You just don't understand". PMID- 12653085 TI - Doctors and managers. Should we make managers more accountable to doctors? PMID- 12653086 TI - Doctors and managers. Problems in Dutch hospitals resemble those in British hospitals. PMID- 12653087 TI - Values and leadership. Chief executives need to participate in conversation on values and live them. PMID- 12653088 TI - Values and leadership. Global leadership is in disarray. PMID- 12653089 TI - [Effects of uraemia and its treatment on respiratory function]. PMID- 12653090 TI - The making of a disease: female sexual dysfunction. Diversity of experiences should be acknowledged. PMID- 12653091 TI - The making of a disease: female sexual dysfunction. Conspiracy of silence hinders understanding. PMID- 12653092 TI - Protocol #0207-545: a phase I/II, escalating dose, open-label evaluation of safety, feasibility, and tolerability of transgenic lymphocyte immunization (TLI) vaccine subjects with histologically proven prostate adenocarcinoma. PMID- 12653093 TI - The making of a disease: female sexual dysfunction. Inaccuracies are not helpful. PMID- 12653094 TI - [When is a dialysis patient hyporesponsive to erythropoietin?]. PMID- 12653095 TI - Protocol#0207-546: a phase I/II, double-blind, randomized, placebo-controlled study to assess the safety and efficacy of AMG0001 to improve perfusion in critical leg ischemia. PMID- 12653096 TI - Protocol #0207-544: a phase study to evaluate the safety and pharmacokinetics of Pro-1, a liposome-encapsulated thymidine kinase gene formulation, in patients with stage IV metastatic melanoma. PMID- 12653099 TI - Sensory stimulation in dementia. Bright light may not illuminate. PMID- 12653100 TI - Privacy rule shines spotlight on HIM expertise. PMID- 12653101 TI - Audit trails: HIPAA's system of checks and balances. PMID- 12653103 TI - Leading the mad dash to HIPAA compliance. Interview by Jessica Squazzo. PMID- 12653102 TI - ARLNA in the 1930s: the stars beckon onwards. PMID- 12653104 TI - Awareness under TIVA. PMID- 12653105 TI - Protective effect of N-acetylcysteine on renal ischemia/reperfusion injury in the rat. AB - BACKGROUND: Oxygen free radicals are important components involved in the pathophysiological tissue alterations observed during ischemia/reperfusion (I/R). METHODS: The protective effect of N-acetylcysteine (NAC) against the damage inflicted by reactive oxygen species during renal I/R was investigated in Wistar Albino rats using biochemical parameters. Animals were unilaterally nephrectomized, and subjected to 45 min of renal pedicle occlusion followed by lh of reperfusion. N-acetylcysteine (150 mg/kg, i.p.) or vehicle was administered twice, 15 min prior to ischemia and immediately before the reperfusion period. At the end of the reperfusion period, rats were killed by decapitation. For biochemical analysis, the lipid peroxidation product malondialdehyde (MDA) and glutathione (GSH) levels, myeloperoxidase (MPO) activity and protein oxidation (PO) were tested. Serum creatinine and BUN concentrations were measured for the evaluation of renal function. RESULTS: I/R induced nephrotoxicity, as evidenced by increases in BUN and creatinine, was reversed by NAC. The decrease in GSH and increases in MDA, MPO and PO induced by I/R indicated that renal injury involves free radical formation. CONCLUSIONS: Since NAC reversed these oxidant responses, and protected rat renal proximal tubules from in vitro simulated reperfusion injury, it seems that NAC protects kidney tissue against oxidative damage. PMID- 12653106 TI - Epidermal growth factor receptor polymorphism and autosomal dominant polycystic kidney disease. AB - BACKGROUND: The clinical variability in the rate of progression of autosomal dominant polycystic kidney disease (ADPKD) has been attributed to genetic heterogeneity, though environmental factors and modifying genes very likely play an important role as well. We examined the association between clinical outcome, defined by age at onset of end-stage renal disease (ESRD) in 46 ADPKD patients, and a polymorphism in the epidermal growth factor receptor (EGFR) gene, a candidate modifying gene. EGFR is a key element in renal tubular proliferation. METHODS: This study comprised 46 unrelated patients with ADPKD and ESRD, and 58 healthy controls. The patients had prevalently PKD 1 mutations. The EGFR microsatellite polymorphism was genotyped according to Gebhardt et al (11). RESULTS: The allele frequencies of the EGFR polymorphism were different in the ADPKD sample and the control population (G2=17.19; P=0.009). In particular, the frequencies of the 122 and 118bp length alleles had a different distribution (P=0.010 and P=0.047 respectively). Patients with the 122bp length polymorphism had ESRD at an earlier age,but this finding was not statistically significant. CONCLUSIONS: These findings suggest an association between the EGFR microsatellite polymorphism and ADPKD. However, it is difficult to establish which alleles are protective and which harmful. A larger, multicenter study may help clarify these results and is also required to replicate our preliminary finding of an association between ADPKD and the EGFR polymorphism. PMID- 12653107 TI - Class V lupus nephritis: a clinicopathologic study in 152 patients. AB - BACKGROUND: Class V lupus nephritis (LN) can be divided into two subgroups according to the 1995 WHO modified classification, but the difference in clinical characteristics between these subgroups is not well known. METHODS: We classified 152 patients with Class V LN, confirmed by renal biopsy, into two subgroups (61 Class Va, 91 Class Vb), and enrolled 488 patients with Class IV as controls. The clinical manifestations, serologic results and prognosis were compared for Classes Va and Vb. RESULTS: The incidence of hypertension and anemia in Class Vb patients was significantly higher than in Class Va (38.5% vs 21.3%, 72.5% vs 52.5%, p<0.05). The incidence of hematuria and renal insufficiency in Class Vb was 64.8%and 15.4%, which was higher than Class Va (44.2% and 3.3%), but lower than Class IV (89.1% and 35%), p<0.05. The percentage of patients with positive anti-dsDNA antibody and hypocomplementemia in Class Vb tended to be higher than Class Va (35.2% vs 26.2%, 50.6% vs 31.2%). Repeated renal biopsies in 24 patients (11 Class Va, 13 Class Vb) showed that eight Class Vb patients had "transformed" to Class IV LN, while only two Class Va patients did (p<0.05). In three Class Va patients serum creatinine doubled during follow-up, but none of them progressed to end-stage renal disease (ESRD). In Class Vb serum creatinine doubled in ten patients, and three progressed to ESRD. CONCLUSIONS: The renal injury and extrarenal manifestations of Class Vb patients were severer than Class Va. Class Vb patients were more likely to shift to Class IV LN, and the prognosis was poorer than for Class Va. PMID- 12653108 TI - Tidal volumes in ARDS and meta-analysis. PMID- 12653109 TI - Tidal volumes in ARDS and meta-analysis. PMID- 12653110 TI - Tidal volumes in ARDS and meta-analysis. PMID- 12653111 TI - Thiamin regulates agglutination and zygote formation in Schizosaccharomyces pombe. AB - Nutritional conditions regulate mating of the fission yeast S. pombe. To investigate how nutritional signals are monitored by the cell and translated into appropriate mating behaviour, effects of unique and specific growth factors would be desirable. We show that thiamin can inhibit sexual agglutination and zygote formation in S. pombe. A concentration of 50 nM thiamin in the culture medium is required for full growth of a thiamin auxotrophic strain. At this concentration thiamin starts to inhibit mating of wild-type cells of opposite heterothallic mating type and at a 1 microM concentration zygote formation is inhibited by more than 95%. Growth conditions modulate the inhibitory effect of thiamin. Thiamin acts only for a restricted period of time and seems to inhibit commitment to zygote formation rather than the cell aggregation and fusion process itself. Pyrithiamin, a thiamin antagonist, inhibits growth as well as mating. PMID- 12653112 TI - Vacuum assisted wound closure therapy. AB - Vacuum assisted closure (VAC) therapy is designed to promote the formation of granulation tissue for faster healing in the wound beds of patients with acute and chronic wounds. Four controlled trials and one interim analysis provide poor quality data and weak evidence that VAC therapy may be superior to conventional methods used in healing wounds. Complications with VAC therapy are uncommon. Studies are needed to evaluate the effectiveness of VAC therapy when the types of dressings are the same for patients in the groups being compared and VAC therapy is the only differing intervention. PMID- 12653113 TI - Medical devices; reclassification of the knee joint patellofemorotibial metal/polymer porous-coated uncemented prosthesis and the knee joint femorotibial (unicompartmental) metal/polymer porous-coated uncemented prosthesis. Final rule. AB - The Food and Drug Administration (FDA) is announcing that it has reclassified two fixed-bearing knee joint prostheses, the knee joint patellofemorotibial metal/polymer porous-coated uncemented prosthesis, which is intended to be implanted to replace a knee joint, and the knee joint femorotibial (uni compartmental) metal/polymer porous-coated uncemented prosthesis, which is intended to be implanted to replace part of a knee joint. FDA has reclassified the devices from class III (premarket approval) into class II (special controls). The special control that will apply is a guidance document entitled "Class II Special Controls Guidance Document: Knee Joint Patellofemorotibial and Femorotibial Metal/Polymer Porous-Coated Uncemented Prostheses; Guidance for Industry and FDA." The agency is reclassifying these devices into class II because special controls, in addition to general controls, will provide reasonable assurance of the safety and effectiveness of the devices, and there is sufficient information to establish special controls. The agency is also announcing that it has issued an order in the form of a letter to the Orthopedic Surgical Manufacturers Association (OSMA) reclassifying the devices. PMID- 12653114 TI - Gastric lymphoma: is the worldwide incidence rising? Comment. PMID- 12653115 TI - Functioning dialysis fistulae. PMID- 12653116 TI - Clinical characteristics, evaluation, treatment and prognosis of spontaneous coronary dissection. PMID- 12653117 TI - When doctors go to class, industry often foots the bill: lectures tend to feature pills made by course sponsors; companies deny influence; a purple heartburn brochure. PMID- 12653118 TI - Can pesticide tests on humans ever meet standards for ethics? PMID- 12653119 TI - Reports of cloned baby overshadow subtle shift; increasing number of ethicists support process for research. PMID- 12653120 TI - The real tragedy of the commons. PMID- 12653121 TI - Confidentiality in end-of-life and after-death situations. AB - Confidentiality is one of the foundations on which psychotherapy is built. Limitations on confidentiality in the therapeutic process have been explained and explored by many authors and organizations. However, controversy and confusion continue to exist with regard to the limitations on confidentiality in situations where clients are considering their options at the end of life and after a client has died. This article review these 2 areas and provides some suggestions for future research. PMID- 12653122 TI - Applied ethics in mental health in Cuba: part I--guiding concepts and values. AB - As part of a project on professionals' lived experience of ethics, this article explores the guiding concepts and values concerning ethics of mental health professionals in Cuba. The data, obtained through individual interviews and focus groups with 28 professionals, indicate that Cubans conceptualize applied ethics in terms of its central role in professional practice and its connection to the social context and subjective processes. Findings also show that Cuban professionals are guided not only by a set of professional values but by a set of civic values as well. The former are subdivided into other-oriented values and self-oriented values. The study of ethics in another culture such as Cuba offers a unique point of view from which to critique the social construction of our own conceptions of applied ethics in North America. PMID- 12653124 TI - Vaccines versus vectors. PMID- 12653123 TI - Applied ethics in mental health in Cuba: part II--power differentials, dilemmas, resources, and limitations. AB - This article is the second one in a series dealing with mental health ethics in Cuba. It reports on ethical dilemmas, resources and limitations to their resolution, and recommendations for action. The data, obtained through individual interviews and focus groups with 28 professionals, indicate that Cubans experience dilemmas related to (a) the interests of clients, (b) their personal interests, and (c) the interests of the state. These conflicts are related to power differentials among (a) clients and professionals, (b) professionals from different disciplines, and (c) professionals and organizational authorities. Resources to solve ethical dilemmas include government support, ethics committees, and collegial dialogue. Limitations include minimal training in ethics, lack of safe space to discuss professional disagreements, and little tolerance for criticism. Recommendations to address ethical dilemmas include better training, implementation of a code of ethics, and provision of safe space to discuss ethical dilemmas. The findings are discussed in light of the role of power in applied ethics. PMID- 12653125 TI - Prevention of tick-borne diseases: the need for a revised strategy. PMID- 12653126 TI - Rabies epizootics among raccoons vary along a North-South gradient in the Eastern United States. AB - The characteristics of rabies epizootics among raccoons were investigated in 11 eastern states along a North-South gradient from New York to North Carolina. Epizootics were defined as discrete intervals of time of at least 5 months in duration, when reported cases of raccoon rabies from an individual county exceeded the median value of raccoon rabies cases reported by that county over the entire period rabies was present among raccoons in the county. Over the approximately 20-year study period, 35,000 cases of raccoon rabies were reported, and epizootics were detected from 251 (64.4%) of 390 counties. The median annual incidence was 0.14 epizootics per year. During the first defined epizootic in a county, the median total number of raccoons reported rabid was 47, with a median monthly incidence of rabies in raccoons of 3.1. The median lag time from the first report of a rabid raccoon in a county to the beginning of the first epizootic was 4 months. Significant differences in the annual incidence of epizootics and monthly incidence of rabid raccoons during epizootics were observed among different states. Although human population density and per capita health spending within counties were positively associated with increasing magnitude of epizootics, a significant difference in the characteristics of rabies epizootics in northern and southern states was apparent. We hypothesize that environmental conditions and perhaps human influence resulted in rabies epizootics in southern states that were smaller, less-frequent, and lacking in well-defined temporal structure compared with those in northern states. PMID- 12653127 TI - Lassa fever in Guinea: I. Epidemiology of human disease and clinical observations. AB - The arenavirus Lassa is found in West Africa, where it sometimes causes a severe illness called Lassa fever. Lassa fever has been seldom investigated outside of a few hyperendemic regions, where the described epidemiology may differ from that in areas of low or moderate incidence of disease. Through a prospective cohort study, we investigated the epidemiology and clinical presentation of Lassa fever in Guinea, where the disease has been infrequently recognized. A surveillance system was established, and suspected cases were enrolled at five Guinean hospitals. Clinical observations were made, and blood was taken for enzyme-linked immunosorbent assay testing and isolation of Lassa virus. Lassa fever was confirmed in 22 (7%) of 311 suspected cases. Another 43 (14%) had Lassa IgG antibodies, indicating past exposure. Both sexes and a wide variety of age and ethnic groups were affected. The disease was more frequently found, and the IgG seroprevalence generally higher, in the southeastern forest region. In some areas, there were significant discrepancies between the incidence of Lassa fever and the prevalence of antibody. Clinical presentations between those with Lassa fever and other febrile illnesses were essentially indistinguishable. Clinical predictors of a poor outcome were noted, but again were not specific for Lassa fever. Case-fatality rates for those with Lassa fever and non-Lassa febrile illnesses were 18% and 15%, respectively. Seasonal fluctuation in the incidence of Lassa fever was noted, but occurred similarly with non-Lassa febrile illnesses. Our results, perhaps typical of the scenario throughout much of West Africa, indicate Lassa virus infection to be widespread in certain areas of Guinea, but difficult to distinguish clinically. PMID- 12653128 TI - Lassa fever in Guinea: II. Distribution and prevalence of Lassa virus infection in small mammals. AB - Rodents of the genus Mastomys form the reservoir for Lassa virus (LV), an arenavirus that causes a potentially severe hemorrhagic illness, Lassa fever (LF). Although Mastomys rodents exist throughout sub-Saharan Africa, areas of human LF appear to be quite focal. The distribution of small mammals and LV infected Mastomys has been assessed in only a few countries. We conducted a survey of small mammals in selected regions of Guinea to assess the degree to which LV poses a public health risk in that country. A total of 1,616 small mammals, including 956 (59%) Mastomys, were captured from 444 households and seven bush sites. Mastomys made up > 90% of the captured animals in the savannah, savannah-forest transition, and forest regions of Guinea, while Mus musculus dominated in coastal and urban sites. Animals were analyzed via enzyme-linked immunosorbent assay (ELISA) for LV-specific antigen (blood and spleen homogenate) and IgG antibody (blood only). Virus isolation from spleen homogenates was also performed on a subset of animals. Lassa antibody and antigen were found in 96 (11%) and 46 (5%), respectively, of 884 tested Mastomys. Antibody and antigen were essentially mutually exclusive and showed profiles consistent with vertical transmission of both LV and antibody. LV was isolated only from Mastomys. ELISA antigen constituted an acceptable surrogate for virus isolation, with a sensitivity and specificity when performed on blood of 78% (95% confidence interval: 68-83%) and 98% (95-99%), respectively. The proportion of LV-infected Mastomys per region ranged from 0 to 9% and was highest in the savannah and forest zones. The proportion of infected animals per village varied considerably, even between villages in close proximity. Infected animals tended to cluster in relatively few houses, suggesting the existence of focal "hot spots" of LV infected Mastomys that may account for the observed heterogeneous distribution of LF. PMID- 12653129 TI - Simulation studies of St. Louis encephalitis virus in south Florida. AB - Two simulation models were used to investigate the epidemiology of St. Louis encephalitis virus (SLEv) in south Florida, one including sentinel hosts (chickens) and amplification hosts (wild birds), while the other one included age structure in the amplification host population. The overall population size of the vector, Culex nigripalpus, was a major factor in the likelihood of epizootics for both models, but the seasonal dynamics of the vector alone did not explain variation in transmission. Interactions between seasonal factors in the mosquito and reproduction in the wild amplification avian hosts appeared to be important in the likelihood of epizootics. Biased feeding between sentinel and amplification hosts affected the time course of virus prevalence and may have implications for the interpretation of sentinel data. The time of virus introduction strongly affected the timing of outbreaks but did not affect the likelihood of epizootics. In most cases, the outbreak occurred immediately after virus introduction; however, in some cases the outbreak was delayed until the mosquito population increased. This has implications for the timing of control strategies directed against mosquito populations. PMID- 12653130 TI - Simulation studies of St. Louis encephalitis and West Nile viruses: the impact of bird mortality. AB - West Nile virus (WNv) has spread through much of the eastern United States following its introduction in 1999, and arrived in Florida in 2001. Prior to its arrival, we anticipated that its transmission cycle was likely to be similar to that of St. Louis encephalitis virus (SLEv). However, high levels of avian mortality have been reported for WNv in the northeastern United States, and it was unknown how this would impact the transmission dynamics of WNv. Simulation models were used to compare the two viruses by considering the impact of bird mortality on the transmission dynamics of arboviruses in south Florida. Transmission models without disease-induced mortality (SLEv) were compared with models including disease-induced mortality (WNv). Disease-induced mortality depressed transmission, eliminating epizootics in two of 14 simulations that were epizootic without the additional mortality. In both models, the most important factor in the likelihood of epizootics was mosquito population size; the mosquito mortality rate was also important. The additional avian mortality altered the factors most important in the size and timing of epizootics, although it did not always directly affect the outcome of the simulations. In some cases, low-level transmission occurred prior to the epizootic peak. When disease-induced avian mortality was included in the simulations, appreciable numbers of dead birds occurred prior to high levels of infection in mosquitoes. This has implications for the use of dead birds as a surveillance tool monitoring the spread and transmission of WNv. PMID- 12653131 TI - Detection of Borrelia duttonii, a tick-borne relapsing fever agent in central Tanzania, within ticks by flagellin gene-based nested polymerase chain reaction. AB - Argasid ticks collected in the site near Mvumi Hospital, Dodoma, Tanzania, were subjected to flagellin gene-based nested polymerase chain reaction (PCR) amplification for examination of borrelial infections. Eight of 13 ticks gave a strong 350-bp signal; three had a weak signal at the same size, and the rest were negative. Sequence determination of eight of the positive samples resulted in three types of flagellin gene sequences. The first type of sequence (shown by three individuals) was identical to that of Borrelia duttonii strain Ly. The second type of sequence from PCR products from four individual ticks had only one base substitution without amino acid alteration in deduced protein sequence. The third type of sequence was different from that of any other Old World relapsing fever borreliae, and the tick was thought to be infected with an unknown Borrelia species. Ticks were also examined to determine the nucleotide sequence of the mitochondrial 16S rRNA gene. The partial sequence of approximately 470 bases was aligned for comparison with previously published sequences to identify the species. The sequences of 13 individual ticks were all identical, and the sequence similarity analysis revealed the ticks should be classified as members of the Ornithodoros porcinus species. The PCR method described in this report appears to be a reliable tool for the detection of borreliae and epidemiological study of tick-borne relapsing fever. PMID- 12653132 TI - Issues in public health entomology. AB - Public health entomology focuses on the population biology of vector-borne infections, seeking to understand how such pathogens perpetuate over time and attempting to devise methods for reducing the burden that they impose on human health. As public health entomology passes its centennial, a series of pervasive research themes and spirited debates characterize the discipline, many reflecting a tension between field and laboratory research. In particular, institutional support for population-based research and training programs has fallen behind that for those using modern lab-based approaches. Discussion of modes of intervention against vector-borne infections (such as deployment of genetically modified vectors, the role of DDT in malaria control, host-targeted acaricides for Lyme disease risk reduction, and truck-mounted aerosol spraying against West Nile virus transmission) illustrates the discipline's need for strengthening population-based research programs. Even with the advent of molecular methods for describing population structure, the basis for anophelism without malaria (or its eastern North American counterpart, ixodism without borreliosis) remains elusive. Such methods have not yet been extensively used to examine the phylogeography and geographical origins of zoonoses such as Lyme disease. Basic ecological questions remain poorly explored: What regulates vector populations? How may mixtures of pathogens be maintained by a single vector? What factors might limit the invasion of Asian mosquitoes into North American sites? Putative effects of "global warming" remain speculative given our relative inability to answer such questions. Finally, policy and administrative issues such as the "no-nits" dictum in American schools, the Roll Back Malaria program, and legal liability for risk due to vector-borne infections serve to demonstrate further the nature of the crossroads that the discipline of public health entomology faces at the start of the 21st Century. PMID- 12653133 TI - A relapsing fever group spirochete transmitted by Ixodes scapularis ticks. AB - A species of Borrelia spirochetes previously unknown from North America has been found to be transmitted by Ixodes scapularis ticks. Infected ticks are positive for Borrelia spp. by DFA test but negative for Borrelia burgdorferi by polymerase chain reaction (PCR) using species-specific primers for 16S rDNA, outer surface protein A, outer surface protein C, and flagellin genes. A 1,347-bp portion of 16S rDNA was amplified from a pool of infected nymphs, sequenced, and compared with the homologous fragment from 26 other species of Borrelia. The analysis showed 4.6% pairwise difference from B. burgdorferi, with the closest relative being Borrelia miyamotoi (99.3% similarity) reported from Ixodes persulcatus in Japan. Phylogenetic analysis showed the unknown Borrelia to cluster with relapsing fever group spirochetes rather than with Lyme disease spirochetes. A 764-bp fragment of the flagellin gene was also compared with the homologous fragment from 24 other Borrelia species. The flagellin sequence of B. burgdorferi was 19.5% different from the unknown Borrelia and showed 98.6% similarity with B. miyamotoi. A pair of PCR primers specifically designed to amplify a 219-bp fragment of the flagellin gene from this spirochete was used to survey field collected I. scapularis nymphs from five northeastern states (Connecticut, Rhode Island, New York, New Jersey, and Maryland). Positive results were obtained in 1.9-2.5% of 712 nymphs sampled from four states but in none of 162 ticks collected from Maryland. Transovarial transmission was demonstrated by PCR of larval progeny from infected females with filial infection rates ranging from 6% to 73%. Transstadial passage occurred from larvae through adults. Vertebrate infection was demonstrated by feeding infected nymphs on Peromyscus leucopus mice and recovering the organism from uninfected xenodiagnostic larvae fed 7-21 days later. Considering the frequency of contact between I. scapularis and humans, further work is needed to determine the potential public health significance of yet another zoonotic agent transmitted by this tick species. PMID- 12653134 TI - An analysis of spirochete load, strain, and pathology in a model of tick transmitted Lyme borreliosis. AB - Four laboratory-grown, low-passage isolates of Borrelia burgdorferi sensu stricto, B31, JD-1, 910255, and N40, were incorporated into Ixodes scapularis ticks to examine the pathogenesis of these isolates in mice after tick transmission. All isolates induced multifocal, lymphoid nodular cystitis, subacute, multifocal, necrotizing myocarditis, and a localized periostitis and arthritis of the femorotibial joint 6-18 weeks after tick infestation. In terms of the number of mice that demonstrated pathology in bladder, heart, and joint, the highest incidence of lesions occurred 12 weeks after tick bite. Utilizing the Taqman quantitative polymerase chain reaction (q-PCR) fluorogenic detection technology to amplify a conserved region of the flagellin gene, a trend was demonstrated between the number of spirochetes in tissue with duration of pathology. The q-PCR assay developed for this study was sensitive and could reliably measure as few as 1 to 10 spirochetes in the target tissues tested. A higher percentage of B31- and N40-infected mice (92 and 100%, respectively) developed myocarditis than JD-1- or 910255-infected mice (67 and 46%, respectively) 12 weeks after tick bite. The amount of spirochetal DNA that could be amplified for heart at this time point was not statistically different between isolates, indicating a difference in virulence between B31 and N40 relative to JD 1 and 910225. N40-infected mice demonstrated a significantly higher spirochete load (an average of 1.23 spirochetes/mg of tissue, p = 0.045) in femorotibial joints 18 weeks after infection, with 60% of these mice maintaining lesions compared with those infected with B31 (13%), JD-1 (25%), or 910255 (50%), which averaged <0.5 spirochetes/mg of tissue. This mouse model of Lyme borreliosis, including the ability to monitor lesion development and spirochete load, can facilitate the testing of therapeutic regimens for the later stages of tick transmitted Lyme disease and help investigate aspects of the immunopathogenesis of lesion development. PMID- 12653135 TI - Infection and transovarial transmission of rickettsiae in Dermacentor variabilis ticks acquired by artificial feeding. AB - In this study we examined the efficiency of an in vitro feeding technique using glass microcapillaries as a method of establishing rickettsiae-infected lines of ticks. To quantify the volume ingested by ticks during microcapillary feeding, the incorporation of radiolabeled amino acids in tick gut and hemolymph was calculated. Fifteen of 18 ticks consumed between 0.06 and 6.77 microl. However, ingestion of fluid was not correlated to weight gain during capillary feeding. Uninfected and partially fed laboratory-reared female Dermacentor variabilis ticks were exposed to either Rickettsia montana- or Rickettsia rhipicephali infected Vero cells via microcapillary tubes, returned to rabbit hosts, and allowed to feed to repletion. All tissues collected from ticks allowed to feed overnight on rickettsiae-infected fluids were found to be infected when examined by IFA. When rickettsiae-infected and uninfected capillary-fed ticks were allowed to feed to repletion and lay eggs, no significant differences in mean engorgement weight or fecundity was observed. When we assessed the efficiency of transovarial transmission of rickettsiae by ticks that imbibed rickettsiae-infected cells by polymerase chain reaction (PCR) and IFA, infection was detected by PCR in the eggs from 85% of the ticks exposed to R. montana and 69% of the ticks exposed to R. rhipicephali. Rickettsial genes were not amplified in samples of the uninfected controls. Examination by IFA of egg samples from females exposed to rickettsiae-infected cells identified rickettsiae in 100% of the samples tested, while the uninfected controls were negative. PMID- 12653136 TI - Effects of acorn production and mouse abundance on abundance and Borrelia burgdorferi infection prevalence of nymphal Ixodes scapularis ticks. AB - Risk of exposure to Lyme disease is a function of the local abundance of nymphal Ixodes ticks that are infected with the etiological agent, the spirochete Borrelia burgdorferi. We monitored abundance of white-footed mice (the principal B. burgdorferi reservoir in the eastern and central United States) and acorns (a critical food resource for mice), and Ixodes scapularis ticks, as well as ambient temperature (cumulative growing degree days) and growing season precipitation, in a forested landscape of southeastern New York State from 1994 to 2000. We found that acorn production in autumn strongly influenced abundance of white-footed mice the following summer and that abundance of mice in summer, when larval ticks are active, influenced the abundance of infected nymphs the following year. Consequently, the abundance of infected nymphal ticks can be predicted from acorn production 1.75 years earlier. Monitoring of natural fluctuations in acorn production thus supports results of prior acorn addition experiments that were conducted at small spatial scales. Growing degree days and precipitation either had no significant effect on density of nymphs or marginally increased the explanatory power of models that included acorns or mouse density as independent variables. We conclude that, at our study site in New York, the risk of human exposure to Lyme disease is affected by mouse density in the prior year and by acorn production 2 years previously. PMID- 12653137 TI - OspA immunization decreases transmission of Borrelia burgdorferi spirochetes from infected Peromyscus leucopus mice to larval Ixodes scapularis ticks. AB - Recombinant outer surface protein A (OspA) vaccination of wild animal reservoirs has potential application for reducing Borrelia burgdorferi transmission in nature and subsequent risk of human infection. As a major reservoir host, the white-footed mouse (Peromyscus leucopus) is a candidate for a vaccination program designed to reduce infection prevalence in vector ticks. In this study we characterized the effect of various levels of immunization with recombinant OspA glutathione transferase fusion protein on transmission dynamics from infected P. leucopus to larval ticks. Control mice were vaccinated with glutathione transferase alone. All mice were experimentally infected with B. burgdorferi before vaccination. The immune responses of the immunized mice were assessed by enzyme-linked immunosorbent assay for antibodies to OspA. Transmission of B. burgdorferi from infected mice was determined by xenodiagnosis with uninfected larval ticks. Spirochetes in ticks were counted by direct immunofluorescence assay. The concentration of antibody to OspA increased with each OspA vaccination but most markedly after the first and second vaccinations. In comparison with control mice, there was reduced transmission by OspA-vaccinated mice to uninfected ticks. One, two, or three doses of OspA reduced infection prevalence in xenodiagnostic ticks by 48%, 92%, or 99% and the numbers of spirochetes per tick by 84%, 98%, or 99%, respectively. This study suggests that vaccination of P. leucopus with OspA could reduce transmission to the tick vector in nature despite prior infection of the reservoir host. PMID- 12653138 TI - Hantavirus transmission: potential role of ectoparasites. AB - Hantavirus pulmonary syndrome is an often-fatal disease thought to be transmitted exclusively by rodents. We report the first evidence of hantavirus-specific RNA (Bayou) from two trombiculid mites (chiggers) and an ixodid tick parasitizing wild-caught rodents at a field site in Texas and also from a trombiculid mite in the free-living predatory stage of the chigger life cycle collected from the same site. PMID- 12653139 TI - Dengue and West Nile virus--an interview with Duane Gubler, Sc.D., reported by Vicki Glaser. PMID- 12653140 TI - Wanted: medical entomologist. PMID- 12653141 TI - Urban zoonoses caused by Bartonella, Coxiella, Ehrlichia, and Rickettsia species. AB - The last half of the 20th Century witnessed an increase in the occurrence and recognition of urban zoonoses caused by members of the genera Bartonella, Coxiella, Ehrlichia, and Rickettsia, all traditionally considered to be members of the family Rickettsiaceae. In recent years, new human pathogens (Bartonella elizabethae, Bartonella henselae, and Rickettsia felis) have been recognized in urban environments. Other newly recognized pathogens (Ehrlichia chaffeensis and Ehrlichia phagocytophila in the United States) have sylvan zoonotic cycles but are present in urban areas because their vertebrate hosts and associated ectoparasitic arthropod vectors are able to survive in cities. Still other agents, which were primarily of historical importance (Bartonella quintana) or have not traditionally been associated with urban environments (Rickettsia rickettsii), have been recognized as causes of human disease in urban areas. Some diseases that have traditionally been associated with urban environments, such as rickettsialpox (caused by Rickettsia akari) and murine typhus (caused by Rickettsia typhi), still occur in large cities at low or undetermined frequencies and often go undetected, despite the availability of effective measures to diagnose and control them. In addition, alternate transmission cycles have been discovered for Coxiella burnetii, Rickettsia prowazekii, and R. typhi that differ substantially from their established, classic cycles, indicating that the epidemiology of these agents is more complex than originally thought and may be changing. Factors leading to an increase in the incidence of illnesses caused by these bacteria in urban areas include societal changes as well as intrinsic components of the natural history of these organisms that favor their survival in cities. Transovarial and transstadial transmission of many of the agents in their arthropod hosts contributes to the highly focal nature of many of the diseases they cause by allowing the pathogens to persist in areas during adverse times when vertebrate amplifying hosts may be scarce or absent. Domesticated animals (primarily cats, dogs, and livestock) or commensal rodents [primarily Norway rats (Rattus norvegicus) and house mice (Mus musculus)] can serve as vertebrate amplifying hosts and bring these agents and their ectoparasitic arthropod vectors into direct association with humans and help maintain transmission cycles in densely populated urban areas. The reasons for the increase in these urban zoonoses are complex. Increasing population density worldwide, shifts in populations from rural areas to cities, increased domestic and international mobility, an increase in homelessness, the decline of inner-city neighborhoods, and an increase in the population of immunosuppressed individuals all contribute to the emergence and recognition of human diseases caused by these groups of agents. Due to the focal nature of infections in urban areas, control or prevention of these diseases is possible. Increased physician awareness and public health surveillance support will be required to detect and treat existing urban infections caused by these agents, to determine the disease burden caused by them, to design and implement control programs to combat and prevent their spread, and to recognize emerging or resurging infections caused by members of these genera as they occur. PMID- 12653143 TI - Spatiotemporal variation in a Lyme disease host and vector: black-legged ticks on white-footed mice. AB - We monitored population density of white-footed mice (Peromyscus leucopus), burdens of immature black-legged ticks (Ixodes scapularis) on mice, and infection prevalence of host-seeking ticks on six forest plots in southeastern New York State from 1995 through 1999. Despite densities of mice that fluctuated two orders of magnitude, average larval and nymphal tick burdens per mouse remained remarkably constant. Spatial variability in mouse density and tick burdens was modest. The total number of larval and nymphal ticks that fed on the mouse population each year depended strongly on population density of mice; a steady increase was observed in both mouse density and total tick meals on mice from 1996 through 1999. The result was a steady increase in the infection prevalence of nymphal and adult ticks with the etiological agent of Lyme disease, Borrelia burgdorferi, over this time. We suggest that fluctuations in population density of mice, combined with possible regulation of tick burdens on mice, may influence risk of human exposure to Lyme disease. PMID- 12653142 TI - Fort Chaffee revisited: the epidemiology of tick-borne rickettsial and ehrlichial diseases at a natural focus. AB - A retrospective cohort study was conducted among troops training at Fort Chaffee, Arkansas, from May through June 1997, to identify infections caused by tick-borne pathogens. Serum samples were tested by IFAs for antibodies to selected Rickettsia and Ehrlichia species and by an investigational EIA for spotted fever group Rickettsia lipopolysaccharide antigens. Of 1,067 guardsmen tested, 162 (15.2%) had antibodies to one or more pathogens. Of 93 guardsmen with paired serum samples, 33 seroconverted to Rickettsia rickettsii or spotted fever group rickettsiae (SFGR) and five to Ehrlichia species. Most (84.8%) of the personnel who seroconverted to SFGR were detected only by EIA, and seropositivity was significantly associated with an illness compatible with a tick-borne disease. In addition, 34 (27%) of 126 subjects with detectable antibody titers reported a compatible illness. The primary risk factor for confirmed or probable disease was finding > 10 ticks on the body. Doxycycline use and rolling up of long sleeves were protective against seropositivity. The risk of transmission of tick-borne pathogens at Fort Chaffee remains high, and use of the broadly reactive EIA suggests that previous investigations may have underestimated the risk for infection by SFGR. Measures to prevent tick bite and associated disease may require reevaluation. PMID- 12653145 TI - Bioterrorism. PMID- 12653144 TI - Interference between the agents of Lyme disease and human granulocytic ehrlichiosis in a natural reservoir host. AB - Agents of Lyme disease (Borrelia burgdorferi) and human granulocytic ehrlichiosis (Ehrlichia phagocytophila) are perpetuated in a natural cycle involving the black legged tick (Ixodes scapularis) and its vertebrate hosts. Using I. scapularis nymphs as the mode of infectious challenge, we studied how infection with one pathogen in white-footed mice (Peromyscus leucopus) affects their ability to acquire the other agent and subsequently to infect larvae, which these agents would do in nature. Two groups of mice were infected with either B. burgdorferi or E. phagocytophila. One week later, B. burgdorferi-infected mice were challenged with E. phagocytophila, and E. phagocytophila-infected mice were challenged with B. burgdorferi. Simultaneously, two control groups of uninfected mice were infected with each agent from the same tick cohorts used on the first groups of mice. Uninfected I. scapularis larvae were fed on all mice for xenodiagnosis at weekly intervals lasting 2 months. For the B. burgdorferi challenge, all control and E. phagocytophila-infected mice acquired B. burgdorferi. However, fewer xenodiagnostic larvae acquired B. burgdorferi from mice with mixed infections compared with mice infected with B. burgdorferi only. For the E. phagocytophila challenge, all five control mice acquired E. phagocytophila, but only two of five mice infected with B. burgdorferi subsequently acquired E. phagocytophila. Consequently, mice with both infections produced fewer xenodiagnostic ticks infected with E. phagocytophila than mice infected with E. phagocytophila only. Thus, a primary infection with either B. burgdorferi or E. phagocytophila in mice inhibited transmission of a second agent, suggesting interference between these two agents. PMID- 12653147 TI - Serologic evidence for West Nile virus infection in birds in Staten Island, New York, after an outbreak in 2000. AB - After an outbreak of West Nile virus (WNV) infections in people, horses, and wildlife in Staten Island, NY, during the summer of 2000, we surveyed the bird population of the island for evidence of infection. Neutralizing antibodies were detected in 59 of 257 (23.0%) resident birds and none of 96 transient (migrating) birds sampled in early October. Species with the greatest seroprevalence were northern cardinal (Cardinalis cardinalis) (69.2%) and rock dove (Columba livia) (54.5%). House sparrows (Passer domesticus) and chickens (Gallus gallus) had lower than expected seroprevalences, 8.6% and 5.5%, respectively. The geographic distribution of seropositivity suggested focal transmission at several locations on the island. The concentration of seropositive birds among resident bird populations on Staten Island supports the concept that many birds survive WNV infection and that some of these play an important role in the WNV-bird-mosquito transmission cycle. PMID- 12653146 TI - Hantavirus pulmonary syndrome in the State of Sao Paulo, Brazil, 1993-1998. AB - Between 1993 and 1998, 10 cases of clinical hantavirus infection were diagnosed in Brazil. Hantavirus-specific IgM, or positive immunohistochemical analysis for hantavirus antigen, or positive reverse transcription-polymerase chain reaction results for hantavirus RNA were used to confirm nine of these cases; eight were hantavirus pulmonary syndrome (HPS), and one was mild hantavirus disease. The remaining clinical case of hantavirus infection was fatal, and no tissue was available to confirm the diagnosis. During the first 7 months of 1998, five fatal HPS cases caused by a Sin Nombre-like virus were reported from three different regions in the State of Sao Paulo, Brazil: two in March (Presidente Prudente Region), two in May (Ribeirao Preto Region), and one in July (Itapecerica da Serra Region). Epidemiologic, ecologic, and serologic surveys were conducted among case contacts, area residents, and captured rodents in five locations within the State of Sao Paulo in June of 1998. Six (4.8%) of 125 case contacts and six (5.2%) of 116 area residents had IgG antibody to Sin Nombre virus (SNV) antigen. No case contacts had a history of HPS-compatible illness, and only one area resident reported a previous acute respiratory illness. A total of 403 rodents were captured during 9 nights of trapping (1969 trap nights). All 27 rodents that were found to be positive for IgG antibody to SNV antigen were captured in crop border and extensively deforested agricultural areas where four of the 1998 HPS case-patients had recently worked. The IgG antibody prevalence data for rodents suggest that Bolomys lasiurus and perhaps Akodon sp. are potential hantavirus reservoirs in this state of Brazil. PMID- 12653148 TI - Acarologic risk of exposure to emerging tick-borne bacterial pathogens in a semirural community in northern California. AB - An acarologic study was conducted in a semirural community in northern California to determine the relative abundance of, and the prevalence of infection with, three emerging bacterial pathogens in the western black-legged tick (Ixodes pacificus). These included the agents causing Lyme disease (Borrelia burgdorferi), human granulocytic ehrlichiosis [Ehrlichia phagocytophila (formerly Ehrlichia equi)], and human monocytic ehrlichiosis (Ehrlichia chaffeensis). The study area in Sonoma County consisted of two properties each with four residents and an uninhabited adjacent comparison area. Six of the eight residents had been either physician-diagnosed or serodiagnosed previously with Lyme disease, and, of these, one also had been serodiagnosed with human monocytic ehrlichiosis. Direct immunofluorescent/culture assays and bacterial species-specific polymerase chain reaction assays were used to test whole ticks individually for presence of B. burgdorferi and Ehrlichia spp., respectively. Overall, 6.5% of the nymphal (n = 589) and 1.6% of the adult ticks (n = 318) from the same generational cohort were found to contain B. burgdorferi. In contrast, none of 465 nymphs and 9.9% of 202 adults were infected with E. phagocytophila. Excised tissues from another 95 adult ticks yielded a comparable E. phagocytophila infection prevalence of 13.7%. E. chaffeensis was not detected in either nymphal or adult ticks. Using a combination of culture and polymerase chain reaction assays, coinfection of I. pacificus adults with B. burgdorferi and E. phagocytophila was demonstrated for the first time. The marked disparity in the infection prevalence of these pathogens in nymphal and adult ticks suggests that their maintenance cycles are inherently different. PMID- 12653149 TI - Risk factors for human exposure to raccoon rabies during an epizootic in Connecticut. AB - The emergence of rabies among terrestrial wildlife poses increasing but poorly defined risks to people. In particular, events leading to human exposure to rabies virus via mammalian reservoirs remain elusive. Thus, we determined those risk factors associated with human exposure to rabies-positive animals during a raccoon rabies epizootic in Connecticut. Existing passive surveillance data on animal rabies tests in Connecticut from 1991 through 1994 were evaluated for demographic, ecological, and behavioral characteristics of human exposure. Of 2,525 rabies-positive terrestrial animals identified, human contact was reported on 556 occasions (22%) and involved at least 939 individuals. The annual incidence of exposure rose from none during 1985-1990 to 66 in 1991 and then averaged 291 during 1992-1994. Exposure was most often indirect in nature, involved a rabies-positive raccoon, was mediated through a domestic animal, and occurred most frequently either near or inside the home. These results suggest that human exposure to rabid animals represents a significant, reemerging public health concern in the United States. Analysis of the epizootiology of rabies infection and of individual exposure risks could reduce inappropriate administration of rabies postexposure prophylaxis, as well as inform other proposed interventions. PMID- 12653150 TI - Characterization of enzootic foci of Venezuelan equine encephalitis virus in western Venezuela. AB - The distribution of the sylvatic subtype ID Venezuelan equine encephalitis (VEE) viruses in the lowland tropical forests of western Venezuela was investigated using remote sensing and geographic information system technologies. Landsat 5 Thematic Mapper satellite imagery was used to study the reflectance patterns of VEE endemic foci and to identify other locations with similar reflectance patterns. Enzootic VEE virus variants isolated during this study are the closest genetic relatives of the epizootic viruses that emerged in western Venezuela during 1992-1993. VEE virus surveillance was conducted by exposing sentinel hamsters to mosquito bites and trapping wild vertebrates in seven forests identified and located by means of the satellite image. We isolated VEE viruses from 48 of a total of 1,363 sentinel hamsters in two of the forests on six occasions, in both dry and wet seasons. None of the 12 small vertebrates captured in 8,190 trap-nights showed signs of previous VEE virus infection. The satellite image was classified into 13 validated classes of land use/vegetation using unsupervised and supervised techniques. Data derived from the image consisted of the raw digital values of near- and mid-infrared bands 4, 5, and 7, derived Tasseled Cap indices of wetness, greenness, and brightness, and the Normalized Difference Vegetation Index. Digitized maps provided ancillary data of elevation and soil geomorphology. Image enhancement was applied using Principal Component Analysis. A digital layer of roads together with georeferenced images was used to locate the study sites. A cluster analysis using the above data revealed two main groups of dense forests separated by spectral properties, altitude, and soil geomorphology. Virus was isolated more frequently from the forest type identified on flat flood plains of main rivers rather than the forest type found on the rolling hills of the study area. The spatial analysis suggests that mosquitoes carrying the enzootic viruses would reach 82-97% of the total land area by flying only 1-3 km from forests. We hypothesize that humans within that area are at risk of severe disease caused by enzootic ID VEE viruses. By contrast, equines could actually become naturally vaccinated, thus preventing the local emergence of epizootic IC VEE virus strains and protecting humans indirectly. PMID- 12653152 TI - Baylisascaris procyonis (Nematoda: Ascaridae) in raccoons (Procyon lotor) in Orange County, California. AB - Baylisascaris procyonis is one of the common nematodes of the raccoon (Procyon lotor), being well recognized as a cause of visceral, ocular, and neural larval migrans in many species of wild and domestic birds and mammals, including humana. Between January 15 and December 31, 2000 in Orange County, CA, 640 raccoons (P. lotor) were live-trapped in four spatial distributions zones, and B. procyonis egg shedding prevalence and fecal egg per gram (EPG) counts were determined in relation to spatial distribution zones, season of the year, age, and sex. Egg shedding prevalence rates were not statistically different between zones, but the EPG count in the primarily residential Zone D differed significantly from other zones. Egg shedding prevalence and EPG counts exhibited a significant rise in the fall and decline in the winter and were significantly higher in juvenile and male raccoons than in adult and female raccoons. The overall fecal egg shedding prevalence was 72.0 +/- 1.8% (461/640), and the overall EPG count was 27,982 +/- 1,006. PMID- 12653151 TI - Estimation of vectorial capacity of Anopheles dirus (Diptera: Culicidae) in a forest-fringed village of Assam (India). AB - To estimate the vectorial capacity of Anopheles dirus, the main vector of forest malaria in the northeastern region of India, in order to gain an understanding of entomological factors related to malaria transmission in forest-fringe areas of Assam, India, an isolated village in the tropical rain forest-fringed area in the district of Dibrugarh, Assam, under the influence of An. dirus alone was studied. Data on various entomological variables required for computation of the vectorial capacity were generated in each month from June 1999 to May 2000 in the field using standard techniques. Malaria prevalence was also studied during the same period in the study village and correlated with the estimated vectorial capacity of An. dirus. Vectorial capacity of An. dirus was highest, 0.779 for Plasmodium vivax (Pv) and 0.649 for Plasmodium falciparum (Pf), during the hot-monsoon season (June-September) and decreased to 0.08 (Pv) and 0.07(Pf) in the temperate postmonsoon season (October-November) before attaining zero values in the cool dry season (December-February). With increasing temperature in the temperate premonsoon season (March-May), vectorial capacity recorded was 0.119 and 0.82 for Pv and Pf, respectively. Significant positive correlation was seen between the estimated vectorial capacity of An. dirus and the number of new Pf (r = 0.86, p < 0.001) and Pv (r = 0.69, p < 0.02) cases in the study village in different months. Thus, this study highlights the pattern of malaria transmission by An. dirus in a forest-fringe area of Assam that begins in March, peaks in July/August, subsides by November, and remains interrupted between December and February. Measures for controlling malaria in forest-fringe areas should be scheduled accordingly. PMID- 12653153 TI - Zoonotic diseases--an interview with Karl M. Johnson, M.D. by Vicki Glaser. PMID- 12653154 TI - Development of allergy post-bone marrow transplantation. PMID- 12653155 TI - Immune responses to allergens early in life: when and why do allergies arise? PMID- 12653156 TI - Pharmacological modulation of human mast cells and basophils. PMID- 12653157 TI - Immune responses to birch in young children during their first 7 years of life. AB - BACKGROUND: The character of immune responses to allergens during the first years of life may decide whether the individual will become tolerant or develop allergy later in life. OBJECTIVE: To study the development of immune responses to the seasonal inhalant allergen birch over the first 7 years of life. METHODS: Blood samples were obtained from 21 children who were followed prospectively from the second to the seventh pollen season of life. Birch-induced cytokine production and IgG subclass antibodies to rBet v 1 were analysed with ELISA, mRNA expression with real time PCR, IgE antibodies to birch with Magic Lite and birch-induced mononuclear cell proliferation with 3H-thymidine incorporation. RESULTS: Birch induced IFN-gamma and IL-10 production increased with age, both in atopic and non atopic children, while birch-induced IL-13 production decreased. The two children who were sensitized and developed clinical allergy to birch showed persistent IL 4 and IL-5 production and IL-9 mRNA expression, as well as Th2-associated IgG4 responses. Transient Th2-like responses were observed among the other children. Proliferative responses and IgG1 antibodies were seen in all children. CONCLUSIONS: Immune responses to birch can be demonstrated in all children, during the first 7 years of life, regardless of atopic status. A transient early Th2-like response is down-regulated after the fourth pollen season, except in children who develop clinical allergy to the particular allergen. PMID- 12653158 TI - Allergy development after bone marrow transplantation from a non-atopic donor. AB - BACKGROUND: Transfer of allergy from atopic bone marrow donors to recipients is known to occur. Development of allergy in a non-atopic patient transplanted from a non-atopic donor is an unfamiliar phenomenon in clinical practice. OBJECTIVES: To clarify the course of events causing a bone marrow recipient to acquire an allergic disease in such non-conducive circumstances. METHODS: Full medical history, prick and intradermal skin tests, and serum IgE levels were obtained from both donor and recipient patients. DNA and red blood cell phenotype analyses were used to detect the degree of chimerism. RESULTS: Only the recipient patient showed positive specific IgE antibodies and skin tests to house dust mite. The recipient patient displayed 100% donor chimera, based on all engraftment markers sought. CONCLUSION: Full engraftment after allogeneic bone marrow transplantation may be associated with modulation of T and B cell function, which in turn could cause the onset of allergic disease after bone marrow transplantation. PMID- 12653159 TI - Is the increase in allergic respiratory disease caused by a cohort effect? AB - BACKGROUND: Changes in lifestyle or environmental factors are responsible for the increasing prevalence of allergic respiratory disease. Establishing the time at which the increase began may provide a clue as to what factors possibly could have contributed to the increase. Many cross-sectional studies have shown that the prevalence of allergic sensitization decreases with increasing age. This could reflect the natural course of allergic sensitization. Alternatively, this could reflect that the increase in sensitization is caused by a cohort effect, i.e. an increase among subjects born during recent decades. OBJECTIVE: The aim was to investigate age-specific changes in the prevalence of allergic sensitization in a cohort of adults. METHODS: A total of 599 subjects aged 15 to 69 years participated in a cross-sectional general population study in 1990. In 1998 they were invited to a follow-up, and 64.4% (386/599) were reexamined. Serum samples obtained from the participants in 1990 and 1998 were analysed for specific IgE to six common inhalant allergens with the same assay. RESULTS: The prevalence of allergic sensitization (specific IgE to at least one allergen) increased among subjects who were less than c. 30 years at baseline (1990), i.e. subjects born during the 1960s or later, while the prevalence was unchanged among subjects who were more than c. 30 years at baseline. CONCLUSIONS: The results support the notion that the increasing prevalence of allergic respiratory disease is caused by a cohort effect. Thus, changes in lifestyle or environmental factors that occurred around or after 1960 may have contributed to this increase. PMID- 12653160 TI - High prevalence of current asthma and active smoking effect among the elderly. AB - BACKGROUND: Although asthma is a common cause of morbidity in adults, relatively few objectively measured population studies of asthma prevalence in adult populations have been conducted. OBJECTIVE: To evaluate the prevalence of asthma, based on both a questionnaire and methacholine bronchial provocation test, and to determine the risk factors of asthma prevalence in an adult population. METHODS: A total of 2,467 adults, who were randomly selected from metropolitan urban, non metropolitan urban and rural areas, responded to the modified ISAAC questionnaire, and underwent methacholine bronchial provocation tests and skin prick tests to locally common aeroallergens. RESULTS: The prevalence of current asthma based on the questionnaire and the methacholine challenge was 2.0% in adults younger than 40, 3.8% in 40- to 54-year-olds, 7.7% in 55- to 64-year-olds and 12.7% in those aged 65 or higher. For subjects of 55-64 years, active smoking was found to be significantly related with the prevalence of current asthma and bronchial hyper-responsiveness, although smoking was positively associated with percentage predictive value of forced expiratory volume of 1 s (FEV1). CONCLUSION: The prevalence of current asthma is common among the elderly, and active smoking may play an important role in the development of asthma and bronchial hyper-responsiveness among the elderly. PMID- 12653161 TI - Specific sensitization to common allergens and pulmonary function in the European Community Respiratory Health Survey. AB - BACKGROUND: The role of atopy in the evolution to chronic obstructive disease remains controversial. AIM: We aimed to assess the association between individual sensitization to common allergens and lung function. METHOD: We analysed data from 12,687 subjects aged 20 to 44 years, from 34 centres in 15 countries participating in the European Community Respiratory Health Survey (ECRHS). Participants performed a blood test, lung function test, methacholine challenge, and answered an administered questionnaire. The relationships between specific IgE, FEV1 and FEV1/FVC ratio were assessed for each study centre stratified by sex, followed by random effects meta-analysis. RESULTS: Asthmatics sensitized to house dust mite had a lower FEV1 (-119 mL in women and -112 mL in men) and FEV1/FVC ratio (-1.95%, and -2.48%) than asthmatics without sensitization. Asthmatics sensitized to cat had a lower FEV1 (statistically significant for women only) and a lower FEV1/FVC ratio. Asthmatic women sensitized to grass had a lower FEV1 and a lower ratio, and those sensitized to Cladosporium had a lower FEV1. A weak association was found with sensitization to cat and to Cladosporium among non-asthmatic women, which disappeared after adjusting for BHR. CONCLUSION: We conclude that atopy was related to a lower lung function, which was only apparent among asthmatics. This relationship was explained by specific sensitization to cat and to house dust mite, the latter being homogeneous across areas. PMID- 12653163 TI - Concentrations of exhaled nitric oxide in asthmatics and subjects with allergic rhinitis sensitized to the same pollen allergen. AB - BACKGROUND: Some studies have reported that the levels of exhaled nitric oxide (ENO) in asthmatics are similar to those in subjects with allergic rhinitis, and it has been postulated that atopic status might be the determinant of enhanced nitric oxide production in asthma. OBJECTIVES: The aim of this study was to determine differences in ENO levels between asthmatics and subjects with allergic rhinitis sensitized to the same allergen, and to correlate these levels with airway responsiveness. METHODS: Nineteen patients with asthma and 18 subjects with allergic rhinitis monosensitized to Parietaria pollen were enrolled in the study. ENO values and airway responsiveness to methacholine and adenosine 5' monophosphate (AMP) were measured during the pollen season. The response to each bronchoconstrictor agent was measured by the provocative concentration required to produce a 20% fall in FEV1 (PC20). ENO was measured with the single-exhalation method. RESULTS: The geometric mean (95% confidence interval) ENO values were significantly higher in asthmatics than in subjects with allergic rhinitis: 72.4p.p.b. (54.9-93.3p.p.b) vs. 44.7p.p.b. (30.9-64.6p.p.b., P = 0.03). In asthmatics, a significant correlation was found between ENO and PC20 AMP values (p = -0.57, P=0.02), whereas no correlation was detected between ENO and PC20 methacholine (p = -0.35, P = 0.14). CONCLUSIONS: Our results suggest that atopy is not the only determinant of increased ENO levels detected in subjects with asthma, and that responsiveness to AMP may be a more sensitive marker for assessing airway inflammation in asthma compared to methacholine. PMID- 12653162 TI - Allergen-induced bronchial inflammation in house dust mite-allergic patients with or without asthma. AB - BACKGROUND: It is presently unknown which factors determine the occurrence and persistence of asthma in house dust mite-allergic individuals. The level of allergen-specific IgE antibodies does not seem to be decisive for asthmatic symptoms. Moreover, levels of exposure to mite allergens do not seem to differ significantly between asthmatic and non-asthmatics individuals. AIM: It was hypothesized that the presence or absence of asthmatic symptoms in house dust mite-allergic patients is associated with quantitative or qualitative differences in the cellular bronchial inflammatory response during the late phase of the allergic reaction. This hypothesis was tested in the bronchial allergen challenge model. MATERIAL AND METHODS: Whole lung challenges with house dust mite extract were performed in 52 house dust mite-allergic subjects, of whom 26 had asthma and 26 had perennial rhinitis without asthmatic symptoms. Primary outcomes were parameters for bronchial inflammation in serial samples of induced sputum (cell differentials, eosinophil cationic protein (ECP), interleukin-8 (IL-8), myeloperoxydase (MPO)). In addition, lung function, non-specific bronchial hyper responsiveness and serial blood samples (eosinophils and IL-5) were analysed. RESULTS: At baseline sputum eosinophils and ECP were similar in both groups but neutrophils and IL-8 were higher in asthmatics. The early bronchoconstriction after allergen challenge was similar in asthma and non-asthmatic rhinitis (median decrease in FEV1: asthma -31.7% vs. non-asthmatics -29.1%, P > 0.1). The late phase bronchoconstriction was significantly greater in asthma (median decrease in FEV1: asthma -27.6% vs. non-asthmatics -18.9%, P = 0.02). Induction of bronchial hyper-responsiveness was similar in both groups. Bronchial allergen challenge elicited significant increases in sputum eosinophils and ECP, which were indistinguishable for both groups (P > 0.1 and P = 0.07, respectively). In contrast, higher numbers of neutrophils persisted in asthma 24h after challenge and were accompanied by significant increases in IL-8 and MPO, which were absent in non-asthmatics (difference between groups P = 0.007 and P = 0.05, respectively). CONCLUSION: Allergen challenge inducedvery similar increases in eosinophils and ECP in induced sputum in allergic asthmatics and in allergic non asthmatic patients. The difference in bronchial inflammation between asthma and non-asthmatic rhinitis appeared to be more closely related to indices for neutrophilic inflammation. PMID- 12653164 TI - Relationship between changes in interferon-gamma production by peripheral blood T cells and changes in peak expiratory flow rate in patients with chronic stable asthma. AB - BACKGROUND: Cytokines production by T helper lymphocytes (Th cells), which orchestrate the interplay of the different cells involved in airway inflammation of asthma, may be reflected in peripheral blood. Some studies have suggested that the Th cell cytokines by peripheral blood T cells correlate with asthma severity. OBJECTIVE: To investigate the relationship between changes in IFN-gamma production by peripheral blood T cells and changes in lung function in chronic stable asthmatics. METHODS: Sixteen patients with chronic stable moderate asthma aged 35-65 years (nine women) were recruited. Morning and evening peak expiratory flow rates (PEFR) monitoring and blood sampling for peripheral blood T cell culture, total IgE and blood eosinophils were performed at baseline and week 12. Levels of IFN-gamma, IL-4 and IL-5 in culture supernatants of peripheral blood T cell were determined by using enzyme-linked immunosorbent assay (ELISA) kits. RESULTS: Patients with increased IFN-gamma changes from baseline showed significantly increased changes in morning (P = 0.02) and evening (P < 0.05) PEFR compared with those with decreased IFN-gamma changes. The changes in IFN-gamma production and IFN-gamma: IL-4 ratio significantly correlated with the changes in morning PEFR (Rs = 0.59, P < 0.02; Rs = 0.63, P < 0.01, respectively) and tended to correlate with the changes in evening PEFR (Rs = 0.45, P = 0.08; Rs = 0.5, P = 0.05, respectively). The changes in IL-4 and IL-5 did not correlate with the changes in IgE and blood eosinophils, respectively. CONCLUSIONS: These findings suggest that IFN-gamma may be associated with the alteration of lung function in asthmatics and play a role in the pathophysiology of chronic stable asthma. PMID- 12653165 TI - Increased aeroallergen-specific interleukin-4-producing T cells in asthmatic adults. AB - BACKGROUND: Asthma, atopy and some forms of respiratory syncytial virus (RSV) disease are thought to be caused by T cells making IL-4 (Th2 cells). However, not all patients with similar patterns of clinical disease have the same underlying pathogenesis and the ability to detect immunopathogenic T cells by examination of the peripheral blood remains in doubt. With the prospect of specific immunotherapy for diseases caused by T cell subsets, it is important to determine whether peripheral blood mononuclear cell (PBMC) reactivity can be used to establish the presence of immunopathogenic responses and therefore to predict therapeutic effects. OBJECTIVE: To detect IL-4 and IFN-gamma production as markers of Th1 and Th2 responses in the peripheral blood of atopic and asthmatic adults. METHODS: PBMC from 22 adult asthmatics (18 of whom were atopic) and 21 non-asthmatic volunteers (ten of whom were atopic) were stimulated with cat, birch and house dust mite allergens, human rhinovirus, RSV and recombinant chimaeric F/G protein from RSV in vitro. ELISPOT assays were used to enumerate cells producing IL-4 and IFN-gamma. RESULTS: Asthmatics had a sixfold increase in frequencies of IL-4-producing cells to cat and birch allergen (median values: 37 vs. 7 per million PBMC, P < 0.01 and 20 vs. 3 per million PBMC, P < 0.04, respectively) compared to non-asthmatics. By contrast, non-asthmatic atopics showed no specific increase in antigen-specific IL-4 responses and there was no evident correlation between skin prick test reactivity and ELISPOT results. Atopics had significantly more IFN-gamma-producing cells specific for FG than nonatopics. while IFN-gamma and IL-4 responses to other antigens were not significantly different. CONCLUSION: Enhanced IL-4 responses to non-viral aeroallergens are seen in adults with asthma, while enhanced IFN-gamma responses to viral antigen FG were see in atopics. In practical terms, ELISPOT assays for specific cytokines may provide a method that could be used to monitor antigen specific T cell responses in peripheral blood. PMID- 12653166 TI - Safety of inhalant allergen immunotherapy with mass units-standardized extracts. AB - BACKGROUND: Allergen-specific immunotherapy (SIT) is an effective treatment for patients with respiratory allergies. However, subcutaneous injection of allergens can provoke systemic side-effects. OBJECTIVE: This study was carried out to determine the incidence and risk factors o fsystemic reactions caused by SIT treatment, using extracts of different inhalant allergens, adsorbed in aluminium hydroxide and biologically standardized with the major allergens quantified in mass units. METHODS: Five hundred and fifty-five subjects with allergic rhinitis and/or asthma were evaluated on clinical history and skin prick test (SPT) reactions to common inhalant allergens. Subcutaneous SIT was administered to all patients, according to the suggested precautionary guidelines and administration schedule. Patients were treated with house dust mite, grass pollen, Parietaria judaica pollen and olive pollen extracts, each receiving one or two extracts. RESULTS: A total of 36,359 injections were administered in the 555 patients. We observed 34 episodes of serious systemic side-effects (0.093% of all injections), in 29 patients (5.2% of all patients), and no fatalities. About 55% of patients reported mild rhinitis and asthma. The majority (59%) of the serious systemic reactions (SSR), and all the anaphylactic reactions, were immediate (i.e. occurred within 30 min after the injection). Asthmatic subjects were at higher risk of SSR than patients with rhinitis (P = 0.01). Most of the side-effects observed occurred during the dose-increase phase (P < 0.05). There was no association of SSR with age, gender, SPT reactivity or allergen type. CONCLUSION: SIT performed in patients with respiratory allergies by specialized staff with the allergen extracts studied, standardized in mass units, provoked a low rate of SSR. The significant risk factors for systemic reactions were asthma and the build-up period. PMID- 12653167 TI - Airway inflammation in thunderstorm asthma. AB - BACKGROUND: Epidemics of acute asthma associated with thunderstorms occur intermittently worldwide, though airway inflammation during these acute episodes has not been characterized. The aim of this study was to characterize airway inflammation in thunderstorm asthma. METHODS: Cases were recruited after presentation to the emergency room with acute asthma immediately following a thunderstorm (n = 6). They were compared to two control groups: a group of atopic asthmatics that had presented with acute asthma to the emergency room prior to the thunderstorm (n = 12), and a second group of corticosteroid naive asthmatics who presented to the emergency room in the prior 12 months (n = 6). Subjects had spirometry, sputum induction and allergy skin tests acutely and at review 4 weeks later. RESULTS: Thunderstorm (TS) cases were more likely to have a history of hay fever and grass pollen allergy, and less likely to be on inhaled corticosteroids (ICS) prior to presentation. Cases and control groups had a similar degree of moderate to severe acute airway obstruction (P = 1.0). TS cases had elevated sputum eosinophils (14.8% of total cell count) compared to controls (1%, 2.6%, P < 0.01). TS cases had higher sputum eosinophil cationic protein (ECP; 11,686 ng/mL) compared to controls (1,883, 3,300, P = 0.02) acutely. TS cases had more cells positive for IL-5 (30%) compared to controls (1, 1.5%, P = 0.02). When adjusted for ICS use, TS cases had a risk ratio for elevated sputum eosinophils of 2.4 (1.23-4.69). CONCLUSION: Thunderstorm asthma is characterized by airway inflammation with IL-5-mediated sputum eosinophilia and eosinophil degranulation. These results are consistent with allergen exposure as the cause of the exacerbation, and are consistent with the thunderstorm-induced grass pollen deluge as the cause of epidemic asthma after thunderstorms. PMID- 12653168 TI - The range of minimum provoking doses in hazelnut-allergic patients as determined by double-blind, placebo-controlled food challenges. AB - BACKGROUND: The risk for allergic reactions depends on the sensitivity of individuals and the quantities of offending food ingested. The sensitivity varies among allergic individuals, as does the threshold dose of a food allergen capable of inducing an allergic reaction. OBJECTIVE: This study aimed at determining the distribution of minimum provoking doses of hazelnut in a hazelnut-allergic population. METHODS: Thirty-one patients with a history of hazelnut-related allergic symptoms, a positive skin prick test to hazelnut and/or an elevated specific IgE level, were included. Double-blind, placebo-controlled food challenges (DBPCFC) were performed with seven increasing doses of dried hazelnut (1 mg to 1 g hazelnut protein) randomly interspersed with seven placebo doses. RESULTS: Twenty-nine patients had a positive challenge. Itching of the oral cavity and/or lips was the first symptom in all cases. Additional gastrointestinal symptoms were reported in five patients and difficulty in swallowing in one patient. Lip swelling was observed in two patients, followed by generalized urticaria in one of these. Threshold doses for eliciting subjective reactions varied from a dose of 1 mg up to 100 mg hazelnut protein (equivalent to 6.4-640 mg hazelnut meal). Extrapolation of the dose-response curve showed that 50% of our hazelnut-allergic population will suffer from an allergic reaction after ingestion of 6 mg (95% CI, 2-11 mg) of hazelnut protein. Objective symptoms were observed in two patients after 1 and 1,000 mg, respectively. CONCLUSION: DBPCFCs demonstrated threshold doses in half of the hazelnut-allergic patients similar to doses previously described to be hidden in consumer products. This stresses the need for careful labelling and strategies to prevent and detect contamination of food products with hazelnut residues. PMID- 12653170 TI - Nasal air sampling used for the assessment of occupational allergen exposure and the efficacy of respiratory protection. AB - BACKGROUND: Occupational exposure to rodent allergens may cause laboratory animal allergy. Personal exposure to occupational allergens is measured by collecting airborne dust on filters using person-carried pumps. This technique cannot be used to evaluate personal protective respiratory equipment. Recently developed intranasal air samplers collect inhaled particles by impaction on adhesive strips within the samplers. OBJECTIVE: The aims were to compare rodent aeroallergen exposure assessment using nasal air samplers with personal air sampling, and to evaluate the efficacy of using respiratory protection during rodent work using nasal air samplers. METHODS: Aeroallergen exposure was assessed during rodent work using both nasal air samplers and personal air samplers. The efficacy of respiratory protection (P2 facemasks and fresh-air helmets) was studied in subject pairs working side by side, one person with protection, the other without. Right nostril samples were laminated with protein-binding membrane and immunostained for rat urinary allergen-containing particles. Left nostril samples and air samples were eluted in buffer and analysed in amplified ELISAs for rat (RUA) and mouse (MUA) urinary allergen content (detection limit 10 pg/mL). Allergen collection efficacy of the nasal air samplers was tested at high and low exposure levels and at different flow rates using static sampling. RESULTS: P2 facemasks decreased the amount of inhaled allergen by about 90%, and verylittle allergen was inhaled using fresh-air helmets. Allergen levels in air and nasal samples correlated well (r(s) was about 0.8 for both RUA and MUA). The number of RUA-positive particles and nasal allergen levels measured in ELISA also correlated significantly (r(s) = 0.8). Collection efficacy of the nasal air sampler was better during high exposure (cleaning cages, median 73% of allergenic particles collected), than during low exposure (undisturbed room, 49% of particles). CONCLUSION: Nasal air sampling is a relevant and sensitive complement to personal air sampling and enables evaluation of personal respiratory protection equipment. Use of P2 facemasks and fresh-air helmets may substantially reduce occupational exposure to inhaled allergens. PMID- 12653169 TI - Zafirlukast has no beneficial effects in the treatment of chronic urticaria. AB - BACKGROUND: Leukotriene receptor antagonists have shown some efficacy in t he treatment of asthma. Injection of LTC4, LTD4 and LTE4 into the skin leads to a weal-and-flare reaction, suggesting an involvement of leukotrienes in the pathogenesis of urticaria. Indeed, various reports have indicated a beneficial effect for leukotriene receptor antagonists in patients with chronic urticaria. OBJECTIVE: To determine the therapeutic effect of the leukotriene receptor antagonist zafirlukast in patients with chronic urticaria. METHODS: The study was a double-blind, placebo-controlled, cross-over study lasting for 12 weeks. Fifty two patients with chronic urticaria were investigated at a university hospital. The patients were randomized to receive 20 mg zafirlukast b.i.d. or placebo and cross-over was scheduled after 6 weeks. The efficacy of the treatment was evaluated by a daily symptom score, six physical examinations, the requirement of rescue antihistamines (acrivastine) and an overall assessment by the patient andthe investigating physician. RESULTS: Forty-six patients completed the study: zafirlukast was well tolerated without alteration of the investigated laboratory parameters. In comparison with placebo, treatment with zafirlukast resulted in no significant positive effect for any of the efficacy measures. Moreover, we were unable to identify any subgroup of patients with chronic urticaria responding with a therapeutic benefit. CONCLUSIONS: The leukotriene receptor antagonist zafirlukast does not provide a significant therapeutic benefit at a dose of 20 mg b.i.d. in patients with chronic urticaria. PMID- 12653171 TI - Comparison of four allergen-sampling methods in conventional and allergy prevention classrooms. AB - BACKGROUND: Several allergen-sampling methods are used to assess level of personal or indirect exposure to cat in homes, schools and other public buildings and working environments. OBJECTIVE: To compare four different allergen-sampling methods (dust collectors, Petri dishes, person-carried pumps and intranasal samplers) by simultaneous sampling in classrooms and to compare the cat allergen levels between conventional classrooms and allergy prevention classrooms. Another aim was to relate the results to self-reported frequency of allergy and asthma symptoms among the children, to their perception of the school environment. METHODS: Among all compulsory schools (n = 257) in the Stockholm suburban area, 35 classrooms (five with implemented allergy prevention measures, seven with additional cleaning and 23 with normal cleaning routines) were chosen for allergen-sampling. Dust collectors (two models), Petri dishes, person-carried pumps and intranasal samplers were used simultaneously. All children (n = 829) received a self-administered questionnaire which included questions about home and school environment, allergic disease, asthma symptoms and pet contact. RESULTS: The correlation between sampling methods was generally poor.Furthermore, there was no significant difference in allergen levels between allergy prevention and allergen avoidance classes compared to conventional classes. Median levels were generally, but not significantly, lower in classes with few cat owners, compared to classes with many cat owners. Children in allergy prevention classes were more satisfied with the indoor air quality and cleaning than children attending classes with fewer or no allergy prevention measures (P < 0.0001). Nine per cent of all children reported allergic symptoms while at school. CONCLUSION: The lack of correlation between sampling methods used simultaneously demonstrates the difficulty in assessing allergen levels in schools and similar environments. The implemented intervention measures (allergy prevention/allergen avoidance) did not influence cat allergen levels at school. PMID- 12653172 TI - Suplatast tosilate inhibits thymus- and activation-regulated chemokine production by antigen-specific human Th2 cells. AB - BACKGROUND: Suplatast tosilate is an anti-allergic agent that suppresses cytokine production by human Th2 cells. OBJECTIVE: We investigated the effects of suplatast tosilate on the production of thymus- and activation-regulated chemokine (TARC) by T cells from allergic patients with asthma. METHODS: Purified protein derivative (PPD)-specific Th1 cell lines and Dermatophagoides farinae (Der f)-specific Th2 cell lines were established from nine patients with house dust mite-allergic asthma. The effects of suplatast tosilate on mRNA expression of TARC and protein production of TARC from antigen-specific Th1 or Th2 cell lines were investigated after stimulation with relevant antigens or phytohemagglutinin (PHA). In addition, the effects of IL-4, IL-10, and IFN-gamma on TARC production by Der f-specific Th2 cell lines in the presence or absence of suplatast tosilate were studied. RESULTS: Although PPD-specific Th1 cell lines did not produce TARC after stimulation with PPD antigen or PHA, stimulation of Der f-specific Th2 cell lines with Der f antigen or PHA increased production of TARC. Suplatast tosilate significantly and dose-dependently inhibited production of TARC by Der f-specific Th2 cell lines stimulated with either Der f antigen (76.5% inhibition at 100 microg/mL, P < 0.01) or PHA (81.9% inhibition at 100 microg/mL, P < 0.01). TARC production by Der f-specific Th2 cell lines was significantly increased only by activation with IL-4 but not with IL-10 or IFN gamma; this increase in TARC production was significantly inhibited by suplatast tosilate (97.5% inhibition at 100 microg/mL, P < 0.01). CONCLUSION: Suplatast tosilate inhibits TARC production by human Th2 cells. Therefore, this agent inhibits both Th2 cytokine and Th2 chemokine and may be a useful anti-allergic agent. PMID- 12653173 TI - Mast cells and T cells in Kimura's disease express increased levels of interleukin-4, interleukin-5, eotaxin and RANTES. AB - BACKGROUND: Kimura's disease (KD) is a chronic inflammatory disorder characterized by tumours in the head and neck region, enlarged lymph nodes, increased eosinophil counts andhigh serum IgE. Mast cells are known to play a central role in IgE-mediated allergic diseases through the release of inflammatory mediators like IL-4, IL-5 and chemokines. We hypothesized that mast cells may play a role in the pathogenesis of KD by regulating eosinophilic infiltration and IgE synthesis. OBJECTIVE: In order to investigate the role of mast cells in the pathogenesis of KD, we examined the expression of cytokines/chemokines in the lesions of KD. METHODS: We examined the number of tryptase+ cells, EG2+ cells, CD3+ cells, IL-4+ cells, IL-5+ cells, eotaxin+ cells, RANTES+ cells and CCR3+ cells in five specimens of KD versus normal tissues by immunohistochemistry. The sources of IL-4, IL-5, eotaxin and RANTES and the expression of CCR3 were examined by immunostaining of serial sections with antibodies to IL-4, IL-5, eotaxin, RANTES and CCR3, and antibodies to tryptase, ECP (EG2) and CD3. RESULTS: Mast cells, activated eosinophils, T cells, IL-4+ cells, IL-5+ cells, eotaxin+ cells, RANTES+ cells and CCR3+ cells were all increased in the lesions of KD as compared with those in normal tissue. Mast cells and T cells were the major source of IL-4, whereas mast cells, T cells and activated eosinophils were the main source of IL-5. Mast cells, T cells and activated eosinophils were the main source of eotaxin and RANTES. CONCLUSIONS: The number of IL-4, IL-5, eotaxin and RANTES-expressing mast cells and T cells were increased in the lesions of KD. As mast cells are lesional resident cells, these cells may play an important role in the pathogenesis of KD by regulating IgE synthesis and orchestrating eosinophilic infiltration. PMID- 12653174 TI - Intranasal application of chitin microparticles down-regulates symptoms of allergic hypersensitivity to Dermatophagoides pteronyssinus and Aspergillus fumigatus in murine models of allergy. AB - BACKGROUND: Previous studies have demonstrated that chitin in the form of microparticles that can be phagocytosed is a potent macrophage stimulator and promotes a Th1 cytokine response and it has been shown that oral administration of chitin microparticles is effective in down-regulating serum IgE and lung eosinophilia in a mouse model of ragweed allergy. To date there have been no studies on the effectivness of directly applying chitin microparticles to the respiratory tract as a treatment for allergic symptoms. OBJECTIVE: To test the effectivness of chitin microparticles when given intranasally as a treatment for the symptoms of respiratory allergy and allergic asthma and to compare its effectivness in two different mouse models of allergy, namely to Dermatophagoides pteronyssinus and Aspergilhus fumigatus. RESULTS: The intranasal application of microgram doses of chitin microparticles is an effective treatment for reducing serum IgE and peripheral blood eosinophilia, airway hyper-responsiveness and lung inflammation in both allergy models results in elevation in Th1 cytokines IL-12, IFN-gamma and TNF-alpha and reduction in IL-4 production during allergen challenge. CONCLUSION: Chitin microparticle suspensions have Th1 immunostimulatory properties and are effective when administered intranasally in mice. The stimulation of the nasal associated lymphoid tissue with chitin microparticles could offer a novel and natural approach to treating allergic disease in humans. PMID- 12653176 TI - The sun and its activity. PMID- 12653175 TI - Depression, quality of life, and lifestyle: chronoecological health watch in a community. AB - The objectives are to explore the possibility of preventive non-drug interventions on vascular disease risk by examining the associations among health related lifestyle (HLS), disease-related illnesses (DRI), subjective quality of life (QOL), depression, and blood pressure (BP). A sample of 181 adults (73 men and 108 women, mean age 57.3 +/- 10.2 years, range 24-76 years) in Urausu, Hokkaido, Japan, wore an ambulatory BP monitor around the clock for seven consecutive days. They completed a health survey questionnaire with which their HLS and DRI were assessed. QOL and depression were rated on the Visual Analogue Scales and the Geriatric Depression Scale-Short Form, respectively. For each participant's systolic (S) and diastolic (D) BP and HR, the circadian MESOR, amplitude, and acrophase were calculated, using cosinor analysis. Associations among the variables were analyzed, using Pearson's correlation coefficient and Kendall's tau-b. DRI was positively associated with depression (P = 0.005) and with HLS (P = 0.001), and was negatively associated with QOL (P = 0.041). Depression showed a moderate and negative correlation with QOL (P < 0.001). As expected, Body Mass Index (BMI) was associated with higher DRI (P = 0.008), SBP (P < 0.001), and DBP (P = 0.002), and with less variation of SBP (P = 0.006) and DBP (P = 0.004). Obesity as assessed by BMI was found to be a good indicator of the circadian BP endpoints and illnesses, warranting further investigation into dietary intake and health outcomes. Depression was also found to be a useful indicator of DRI, HLS, and QOL. PMID- 12653177 TI - The effect of geomagnetic activity on cardiovascular parameters. AB - Multidirectional changes in the natural history of many cardiovascular syndromes have been linked to different levels of daily and monthly geomagnetic activity (GMA). Previous studies have found that in periods of high GMA, there were more admissions for acute myocardial infarction and more cases of anterior wall myocardial infarction. Results also indicated: higher out-patient mortality and a trend towards higher hospital mortality from acute myocardial infarction; higher diastolic arterial pressure in healthy subjects and in treated hypertensive patients; higher prolactin and 17-corticosteroid levels in the peripheral blood; more severe migraine attacks and more admissions for CVA and cerebrovascular insufficiency in male patients; changes in many blood coagulation cellular gradients (platelet count, basophils in the peripheral blood), a rise in platelet aggregation, fibrinogen level and a drop in leukocyte adhesiveness. Periods of low GMA showed a related increase (negative correlation) in in-hospital non myocardial infarction-related cardiovascular deaths. Only in times of lowest GMA did inferior wall myocardial infarction exceed anterior wall myocardial infarction. Low GMA was also associated with higher levels of growth hormone and 11-ketosteroids in the peripheral blood, more sudden deaths, some increase in electrical heart instability number of ventricular and supraventricular extrasystoles and higher rate of ventricular tachycardia. The monthly occurrence of pregnancy-induced hypertension was negatively correlated with GMA level. Gender differences were noted in some of the parameters. Other studied parameters did not show changes related to GMA. These included hemoglobin level, electrolyte level, heart beat and pulse rate. Moreover, some observed cardiovascular fluctuations that were related to the level of GMA, also showed differences in the rising and dropping parts of the 11-year cycle of solar activity. It has been suggested that some of the changes observed in many clinical syndromes may be related to the concomitant activation of the serotoninergic system. PMID- 12653178 TI - Brain-heart connection and the risk of heart attack. AB - Autonomic functions, such as increased sympathetic and parasympathetic activity and the brain's suprachiasmatic nucleus, higher nervous centres, depression, hostility and aggression appear to be important determinants of heart rate variability (HRV), which is, itself, an important risk factor of myocardial infarction, arrhythmias, sudden death, heart failure and atherosclerosis. The circadian rhythm of these complications with an increased occurrence in the second quarter of the day may be due to autonomic dysfunction as well as to the presence of excitatory brain and heart tissues. While increased sympathetic activity is associated with increased levels of cortisol, catecholamines, serotonin, renin, aldosterone, angiotensin and free radicals; increased parasympathetic activity may be associated with greater levels of acetylecholine, dopamine, nitric oxide, endorphins, coenzyme Q10, antioxidants and other protective factors. Recent studies indicate that hyperglycemia, diabetes, hyperlipidemia, ambient pollution, insulin resistance and mental stress can increase the risk of low HRV. These risk factors, which are known to favour cardiovascular disease, seem to act by decreasing HRV. There is evidence that regular fasting may modulate HRV and other risk factors of heart attack. While exercise is known to decrease HRV, exercise training may not have any adverse effect on HRV. In a recent study among 202 patients with acute myocardial infarction (AMI), the incidence of onset of chest pain was highest in the second quarter of the day (41.0%), mainly between 4.0-8.0 AM, followed by the fourth quarter, usually after large meals (28.2%). Emotion was the second most common trigger (43.5%). Cold weather was a predisposing factor in 29.2% and hot temperature (> 40 degrees celsius) was common in 24.7% of the patients. Dietary n 3 fatty acids and coenzyme Q10 have been found to prevent the increased circadian occurrence of cardiac events in our randomized controlled trials, possibly by increasing HRV. We have also found that n-3 fatty acids plus CoQ can decrease TNF alpha and IL-6 in AMI which are pro-inflammatory agents. There is evidence that dietary n-3 fatty acids canenhance hippocampal acetylecholine levels, which may be protective. Similarly, the stimulation of the vagus nerve may inhibit TNF synthesis in the liver and acetylecholine, the principal vagal neurotransmitter, significantly attenuates the release of pro-inflammatory cytokines TNF-alpha, interleukin 1,6 and 18, but not the anti-inflammatory cytokine IL-10 in experiments. Therefore, any agent which can enhance brain acetylecholine levels, may be used as a therapeutic agent in protecting the suprachiasmatic nucleus, higher nervous centres, vagal activity and sympathetic nerve activity which are known to regulate the body clock and HRV and the risk of SCD and heart attack. PMID- 12653179 TI - System times and time horizons for biospheric near-matches of primarily non photic environmental cycles. AB - System time is defined as the duration (T) and sampling density (deltat) of a given study. A time horizon is obtained by combining analytical results on endpoints of time structures (chronomes), usually on control groups, from accumulating diverse studies carried out with drastically different values of T and deltat. These two design considerations depend upon and are limited by resources. The desideratum of a small At e.g., for studies of chaotic endpoints, limits T, e.g., has thus far precluded a continuous mapping of decadal cycles in endpoints such as the approximate entropy or the correlation dimension. Time horizons are being documented for an increasing number of variables that undergo cycles with infra-annual frequencies - with one cycle in about one, two and/or five decades - in the biosphere as in the environment. These infra-annuals, in and around us, modulate ultra-annuals, including circadians, as well as about 7 day (circaseptan) and about half-yearly (circasemiannual) cycles, all prominent, e.g., in geophysical and biospheric specta. Neglect of infra-annual chronomics can lead to blunders. A statistically highly significant decrease in the excretion of steroidal metabolites (or in heart rate variability) may be misinterpreted as premature aging if it lasts for several years, and may be unnecessarily acted upon, e.g., by drugs. The decrease, however, may only be transient, occurring only during a given stage of a circadecadal cycle, and it may be followed for the next several years by an also spurious "rejuvenation", possibly misinterpreted as a drug effect. When recognized as the alternating stages of decadal cycles and assessed with an affordable deltat, infra-annuals of variables involved in major problems of our day, including homicides and wars, may lead us to manipulable internal or external mechanisms, and thus, eventually, possibly to countermeasures to crime and terror as well as to the optimization of aims such as spirituality. PMID- 12653180 TI - The biological effects of solar activity. AB - The synchronization of biological circadian and circannual rhythms is broadly viewed as a result of photic solar effects. Evidence for non-photic solar effects on biota is also slowly being recognized. The ultrastructure of cardiomyocytes from rabbits, the time structure of blood pressure and heart rate of neonates, and the heart rate variability of human adults on earth and in space were examined during magnetically disturbed and quiet days, as were morbidity statistics. Alterations in both the about-daily (circadian) and about-weekly (circaseptan) components are observed during disturbed vs. quite days. The about weekly period of neonatal blood pressure correlates with that of the local geomagnetic disturbance index K. Circaseptans which are seen early in human life and in various other forms of life, including unicells, may provide information about the possible site(s) of life's origins from an integrative as well as adaptive evolutionary perspective. PMID- 12653181 TI - Graded response of heart rate variability, associated with an alteration of geomagnetic activity in a subarctic area. AB - It is becoming recognized that geomagnetic activity may influence biological processes, including the incidence of various human diseases. There is evidence that heart rate variability (HRV) may serve not only as an index of autonomic coordination of the circulation, but also as a powerful predictor of risk in apparently healthy subjects. This study focuses on the effects of geomagnetic disturbance on HRV, by comparing different indices of HRV of young, healthy men living in a subarctic area on days of low (Ap; 0-7), middle (Ap; 7-20), and high (Ap; 20-45) geomagnetic activity. The effect of geomagnetic disturbance on HRV is examined on the basis of 7-day records by Holter ECG, obtained longitudinally on 5 clinically healthy men, 21-31 years of age, in Alta, Norway (70 degree N). Frequency- and time-domain measures of HRV were analyzed for each subject on separate 24-hour spans. A graded alteration of HRV endpoints was found in association with increased geomagnetic activity. As time-domain measures of HRV, SDNNIDX and the 90% length of the Lorenz plot decreased statistically significantly on days with increased geomagnetic disturbance (p = 0.0144 and p = 0.0102, respectively). A graded decrease in frequency-domain HRV measures was also validated statistically for the total spectral power (decrease of 18.1% and 31.6% on days when 7 < Ap < 20 and 20 < Ap < 45 versus days when Ap < 7; p = 0.0013). The decrease in spectral power was mainly found at frequencies below 0.04 Hz, in the "ultra-low-frequency" (0.0001-0.003 Hz; 18.1% and 27.5% decrease, respectively; p = 0.0102) and "very-low-frequency" (0.003-0.04 Hz; 12.9% and 28.6% decrease, respectively; p = 0.0209) regions of the spectrum. The decrease in spectral power was much less pronounced around 10.5 sec ("low frequency"; N.S.) and around 3.6 sec ("high frequency"; N.S.). Evidence is provided here that HRV decreases on magnetically disturbed days, and that it does so in a dose dependent fashion, HRV being depressed more on days when 20 < Ap < 45 than on days when 7 < Ap < 20, by comparison with days when Ap < 7. This graded response of HRV to geomagnetic activity should encourage us to search for human magnetoreceptors and for a better understanding of putative mechanisms of magnetoreception. PMID- 12653182 TI - Is motivation influenced by geomagnetic activity? AB - To eventually build a scientific bridge to religion by examining whether non photic, non-thermic solar effects may influence (religious) motivation, invaluable yearly world wide data on activities from 1950 to 1999 by Jehovah's Witnesses on behalf of their church were analyzed chronobiologically. The time structure (chronome) of these archives, insofar as it is able to be evaluated in yearly means for up to half a century, was assessed. Least squares spectra in a frequency range from one cycle in 42 to one in 2.1 years of data on the average number of hours per month spent in work for the church, available from 103 different geographic locations, as well as grand totals also including other sites, revealed a large peak at one cycle in about 21 years. The non-linear least squares fit of a model consisting of a linear trend and a cosine curve with a trial period of 21.0 years, numerically approximating that of the Hale cycle, validated the about 21.0-year component in about 70% of the data series, with the non-overlap of zero by the 95% confidence interval of the amplitude estimate. Estimates of MESOR (midline-estimating statistic of rhythm, a rhythm (or chronome) adjusted mean), amplitude and period were further regressed with geomagnetic latitude. The period estimate did not depend on geomagnetic latitude. The about 21.0-year amplitude tends to be larger at low and middle than at higher latitudes and the resolution of the about 21.0-year cycle, gauged by the width of 95% confidence intervals for the period and amplitude, is higher (the 95% confidence intervals are statistically significantly smaller) at higher than at lower latitudes. Near-matches of periods in solar activity and human motivation hint that the former may influence the latter, while the dependence on latitude constitutes evidence that geomagnetic activity may affect certain brain areas involved in motivation, just as it was earlier found that it is associated with effects on the electrocardiogram and anthropometry. PMID- 12653183 TI - Non-linear relation of heart rate variability during exercise recovery with local geomagnetic activity. AB - Heart rate variability (HRV) during 30 minute's recovery from a 30-minute exercise was established for 6 healthy men 23-30 years old. The exercise-recovery schedule was performed at 8 circadian stages over 11 days for each subject and analyzed over consecutive 5-minute segments. The local K index for the time of the recovery was recorded as a variable for geomagnetic activity. Pooling all HRV values across the different times into recovery, the relation of HRV to local K is found to be non-linear. A quadratic model fitted to the pooled HRV values is statistically significant. The present study indicates that during recovery from exercise, the effect of geomagnetic disturbances on HRV is non-linear. A non linear relation of HRV with respect to geomagnetic activity deserves further work to determine whether it can account for discrepant findings of an effect of magnetic storms on the incidence of myocardial infarctions, for which a decreased HRV is a predictor. PMID- 12653184 TI - Distribution of monthly deaths, solar (SA) and geomagnetic (GMA) activity: their interrelationship in the last decade of the second millennium: the Lithuanian study 1990-1999. AB - Thirty percent of cardiovascular deaths cannot be explained by known risk factors. In this study, we sought links between 1) circannual rhythmicity, solar activity (SA) - sunspot number, solar radioflux and geomagnetic activity (GMA) - Ap., Cp., Am. with monthly number of deaths looking for environmental influence on mortality at the end of the second millennium. The Lithuanian national death data from for 120 consecutive months according to the IDC (1990 - 1999, n = 424925 deaths) was studied: 157189 from IHD (72144 male, 85045 female), 50228 from stroke (19062 male, 31166 female), 33722 from accidents (26193 male, 7529 female), 10655 from road accidents (8127 male, 2528 female) and 14810 from suicide (12137 male, 2673 female), 217508 - non cardiovascular (134308 male, 83200 female). Monthly death numbers (total, < 65y, 65-74, > 74y old for IHD and Stroke, and by gender) were compared with time of year, SA and GMA indices obtained from the National Geophysical Data and National Space Services Center, USA. Circannual rhythmicity indices, Pearson correlation coefficients and their probabilities were obtained and analyzed. The interrelationships of deaths from IHD and stroke and some other pathologies were also studied. In addition a multivariate linear regression analysis was done in order to evaluate the influence of time (month), Solar (S.A.) and Geomagnetic (GMA) activity on the temporal distribution of deaths. The total IHD monthly death number was significantly linked with GMA indices (r = 0.2-0.24, p = 0.03-0.075), but not with SA. IHD correlated with SA (r = 0.25-0.27, p = 0.006-0.0026), only in the 74 year age group. p = 0.0001) In the 65-74 age group, there was an inverse relationship with SA (r = -0.46-0.44) and no significant relationship to GMA. The IHD death number was correlated with GMA only in the < 65 and > 74 year age group (r = 0.30-0.36, p = 0.001-0.001). 2) Stroke-related deaths showed an inverse correlation with SA and GMA only for the 65-74y age group (r = -0.5, p = 0.0001 - SA, r = -0.4, p = 0.0001- GMA) and with GMA at age < 65 only for males (r = 0.20 0.25, p = 0.03-0.04). 3) Both IHD and stroke were strongly circannual rhythmic with acrophase at month 1.27 (IHD) and 1.32 (stroke) - first half of February, (p = 0.0001). 4) The IHD / stroke death ratio was correlated both with SA and GMA (r = 0.35, p 0.0001 - SA, r = 0.40-0.44, p = 0.0001 - GMA). 5) There was a strong inverse monthly deaths correlation between IHD and suicide (n = 14,810, r = 0.53, p = 0.0001). Stroke and suicide were also related, but to a much weaker extent (r = -0.217, p = 0.017). Accidents were inverse related to S.A. (r = 0.286, p = 0.0015), but not to GMA. Road Accidents - with both S.A. (r = 0.427, p < 0.0001) and GMA (r = 0.258, p = 0.004); with acrophases in October for road and November for other fatal accidents. Non cardiovascular deaths were annually rhythmic only for female - acrophase in early February, and for both gender inverse related to S.A. (r = -0.57, p < 0.0001) but not with GMA. Suicide victims had their acrophase in July (p < 0.0001) and were inverse related to S.A. (r = 0.6, p < 0.0001) and GMA (r = -0.27, p = 0.002). At the onset of the third millennium, the temporal distribution of deaths is still related to environmental physical activity. Those links differ for each of the pathologies, and by age and gender. PMID- 12653185 TI - Changes in circadian rhythm in heart rate and parasympathetic nerve activity after an eastward transmeridian flight. AB - Distortions of the circadian rhythm structure of the heart rate and parasympathetic nerve activity after an eastward transmeridian flight were studied. We recorded the ambulatory electrocardiograms of 12 healthy young men for 11 days: 3 days in Tokyo before the flight and 8 days in San Francisco following it. Heart rate variability was determined by power spectrum analysis, and high frequency power (0.15-0.4 Hz) was used as the index of parasympathetic tone. The structural characteristics of the circadian rhythm were analyzed by dividing consecutive RR data into four parts: the preflight days, the second and third days, the fourth and fifth days (Days 5-6) and the sixth and seventh days. Eight of the 12 cases were analyzed completely. The circadian period of the heart rate decreased after the flight. The peak time of parasympathetic nerve activity shifted from the small hours to the daytime, the maximum shift being during days 5-6 after the flight. We concluded that a structural disturbance of the circadian rhythm of the heart rate and parasympathetic nerve activity were observed. PMID- 12653186 TI - Marriage and divorce over a century in Japan: social biomedicine, not yet societal therapy. AB - During the past half-century, the differences found in the spectral element of marriages vs. divorces in Tokyo reveal a half-year aspect which increasingly dominates in marriages, probably as a result of a mix of socio-economic, climatic and/or geomagnetic, as well as human affection-related factors, On the other hand, a decadal component is prominent in divorces, possibly in association with non-photic solar activity. Sparser (yearly, not monthly) records, obtained over a century at different latitudes in Japan, demonstrate that the decadal components are not consistent in moving spectra of original (as yet not detrended) data. Longer and denser series over much greater differences in latitudes and longitudes will have to be subjected to a much more thorough scrutiny, in conjunction with a series of possibly pertinent variables, to assess the contributions of different sources of variation in different areas of Japan and world wide, preferably in democratic, non-combatant areas, where the socio economical, political and other war-associated conditions are mostly in the background. PMID- 12653187 TI - Geomagnetics and society interact in weekly and broader multiseptans underlying health and environmental integrity. AB - Evidence for the ubiquity and partial endogenicity of about-weekly (circaseptan) components and multiples and/or submultiples thereof (the multiseptans) accumulates as longer and denser records become available. Often attributed to a mere response to the social schedule, circaseptan components now have been documented to characterize environmental variables related to primarily non photic solar effects. Plausibly, like circadians, circaseptans are anchored in genomes, from bacteria to humans, via both an internal and external evolution. If so, circaseptans, like circadians, may be found in the absence of a 7-day schedule, whereas the social schedule may play a synchronizing role and be responsible for the detection of prominent weekly variations in population statistics. The wobbliness of multiseptans and other components of some environmental time structures (chronomes) may correspond to the wobbliness of multiseptans found in cardiovascular morbidity statistics. Here, the latter stem primarily, but not exclusively, from an extensive database on the incidence of daily calls for an ambulance in Moscow, Russia from 1979-1981. A modulation of multiseptans and other chronome components of both environmental and biological variables by the about 11-year solar activity cycle (and of other low-frequency signals reviewed elsewhere) may account for prior controversies and scepticism about a variety of non-photic effects on biota. This is notably the case when relatively short series are analyzed without consideration of effects of unassessed long-term variations; this is the task of the new field of chronomics. In the spectral element of the chronomes of geophysical and biospherical variability, there are natural near weeks,apart from any precise 7-day periodicity. PMID- 12653188 TI - Modulatory influences of ventilatory disorders on electrical stability of the rat heart. AB - It is a known fact that there is a relationship between some ventilatory disorders and the incidence of ventricular arrhythmias. The aim of our study was to show this relation in the circadian dependence in in-vivo rat models. The electrical stability of the heart was measured by ventricular arrhythmia threshold (VAT) at 3h intervals during a 24h period. The experiments were performed in the Wistar rats (pentobarbital anaesthesia 40mg/l kg i.p., open chest experiments, adaptation to the light regime 12:12 hours, with the dark phase from 18.00h to 06.00h). The normal artificial ventilation was used in the control group (n = 17), hypoventilation (n = 10) and hyperventilation (n = 7) in the second and third ones. The fourth group (n = 4) was subjected to 20 min. hypoventilation and subsequent 20 min. reoxygenation. The significant circadian rhythm was detected under normal ventilatory conditions. Hypoventilation significantly decreased (alpha = 0.001) the VAT and changed the 24h rhythm to the moderate biphasic compared to the control group. The biphasic character was evident only after 10 min. of hypoventilation. Hyperventilation non-significantly increased the VAT, but did not change the rhythm. Reoxygenation, after 20 minutes of hypoventilation, expressively changed the VAT circadian rhythm, inversely compared to the control group. The biphasic character was kept only after 5 min. of reoxygenation. It is concluded that myocardial vulnerability to the ventricular arrhythmias is influenced by ventilatory disorders in the circadian dependence. PMID- 12653189 TI - Self-measurement and ambulatory monitoring of blood pressure: a subject's chronobiological perspective. AB - Population studies show the direct benefit from self-measurement for guiding the treatment of patients with severe hypertension; they also separate groups of children with a positive vs. negative family history of high blood pressure, despite the difficulty of obtaining valid readings during the rest/sleep span, which are needed for a reliable estimation of the circadian parameters. From the viewpoint of preventive maintenance, a 67-year-old woman monitored her blood pressure and heart rate around the clock for 7 days, using concomitantly an ambulatory monitor and a manual instrument. Self-measurement profiles were repeated on several occasions. The results are compared with those of a 62-year old woman who followed the same initial 7-day protocol. The estimation of the circadian blood pressure pattern differed between the profiles obtained with an ambulatory monitor and by self-measurement. The differences, relatively small in one subject, were much larger for the other subject. Provided the results are chronobiologically analyzed and interpreted in the light of reference limits which are specific to self-measurement series as well as for gender, age and times of sampling, systematic self-measurement of blood pressure may yield a reliable assessment of the circadian variation for some, but not for all, individuals. For those who have been validated in at least one 7-day/24-h profile, occasional automatic ambulatory profiles are tentatively recommended as the main approach, with complementary systematic self-measurement when automatic instruments are scarce. For those with discrepant results, automatic monitoring is invariably recommended. PMID- 12653191 TI - Oxygen desaturation and heart rate variability due to Cheyne-Stokes respiration in congestive heart failure patients. AB - Cheyne-Stokes respiration is common in congestive heart failure patients and causes cyclical fluctuation of the RR interval. We studied the characteristics of apnea-related heart rate variability (apnea HRV) and the relation between apnea HRV and oxygen desaturation was examined. Ambulatory electrocardiograms and data on respiration (oronasal flow, tracheal sounds, abdominal wall movement and oxygen saturation) were simultaneously recorded by a multi-channel digital recorder for 16 congestive heart failure patients (10 men and 6 women; mean age, 68 +/- 13 years). HRV occurred as a result of cyclical apnea attacks between 0.005 and 0.03 Hz (apnea band). Apnea HRV was obtained as the power ratio of apnea HRV normalized by the very low frequency band (0.003-0.04 Hz). Apnea HRV increased with the number of apnea episodes and the oxygen desaturation index, but no relation between apnea HRV and either mean oxygen density or oxygen desaturation time was observed. We concluded that apnea HRV is a predictor of the number of apnea attacks or oxygen desaturation, but does not reflect the degree of oxygen desaturation in Cheyne-Stokes respiration. PMID- 12653190 TI - Circadian variation of basal total vascular tone and chronotherapy in patients with vasospastic angina pectoris. AB - Vasospastic angina pectoris (VSA) is an anginal attack which occurs characteristically between night and early morning. The aim of this study was to clarify the cause of VSA. The subjects consisted of 16 patients with VSA, 18 patients with effort angina (EAP) and 15 healthy individuals, who were used as the control group. Subjects were attached to an ambulatory blood pressure monitor and a non-invasive continuous cardiac output monitor concurrently, over a 24-hour period. Mean blood pressure (MBP), and cardiac index (CI) were measured. Then basal total vascular tone (TVT) was calculated as follows: basal TVT = (MBP/CI) x 1,332 dyne/sec/cm5. The decrement of CO was greater during sleeping hours as compared with the decrement of the MBP in the VSA group. Nocturnal basal TVT was significantly greater in the VSA group than in the EAP group or the control group. The increased nocturnal basal TVT was significantly suppressed by long acting calcium antagonists to the level of the EAP and the control groups. The treatment also decreased the frequency of ischemic attacks. PMID- 12653192 TI - Effect of H2 blockers on the circadian rhythm of intragastric acidity. AB - Most patients with peptic ulcer or gastro-oesophageal reflux disease develop nocturnal pain (epigastric and retrosternal pain from midnight to early morning), which often disappears before breakfast. Such pain may be related to a disturbance of the circadian rhythm of gastric acid secretion. Helicobacter pylori is a known aetiological agent of peptic ulcer disease and patients with gastritis or ulcers now undergo infection eradication therapy. However, this can result in the onset or exacerbation of gastro-oesophageal reflux disease. There has been a marked increase in the number of patients with oesophagitis rather than peptic ulcer and because most are negative for H. pylori, attention has centred on the status of their gastric acid secretion. Some patients with oesophagitis complain of nocturnal pain despite treatment with a proton pump inhibitor, and in those cases a short course of an H2 blocker can be very effective. We used a portable pH meter to study, in a cross-over fashion, the changes in the circadian rhythm of gastric acid secretion caused by two H2 blockers, laftidine and famotidine, in 10 H. pylori-negative subjects. There was a significant difference in the rhythm between baseline (no treatment) and when laftidine or famotidine were administered, with mean values for amplitude of 28.1, 13.80 and 10.82, respectively; for the midline estimating statistic of rhythm (MESOR), 22.7, 10.80, and 11.54; and for acrophase, 324.0. 312.3, and 274.5 (p < 0.001). The H2 blockers suppressed the normal circadian rhythm of intragastric acidity, which rises in the evening until the middle of the night and then drops in the morning. PMID- 12653193 TI - Intense response of heart rate with pronounced suppression of high-frequency power of heart rate variability to early morning exercise with high-intensity load. AB - Autonomic nerve activity shows circadian variation. Therefore, we put forward the hypothesis that the responses of heart rate (HR) and high-frequency (HF) power of HR variability to exercise would be different between early morning and daytime exercise. We performed ergometer constant load exercise tests [50 watts (low), 100 watts (high load)] in the early morning and during the day in 6 healthy volunteers. The HR response was fitted to an exponential hyperbolic sine function: HR= alpha*e(-beta*t) *sinh(omega*t)+gamma. In this equation, the beta/omega ratio was inversely correlated with the intensity of the HR response. HF power was determined using a recently developed algorithm with high time resolution power. There were no significant differences in HR, HF power or systolic blood pressure (BP) pressure before exercise between early morning and daytime exercise with either the 50 or 100 watt loads. During exercise, there were no significant differences in maximal HR or maximal systolic BP between early morning and daytime exercise with either 50 or 100 watt loads. For high load exercise, the beta/omega ratio was significantly lower in early morning exercise (mean +/- SD, 0.945 +/- 0.02) than in daytime exercise (0.987 +/- 0.03). Similarly, for 100 watt exercise, HF power of HR variability was significantly lower in early morning exercise (0.94 +/- 0.52 msec/Hz 1/2) than in daytime exercise (1.26 +/- 0.74 msec/Hz 1/2). In conclusion, the present study demonstrated that a lower beta/omega ratio in the HR response was associated with lower HF power of HR variability in early morning high-load exercise compared to that in daytime exercise, indicating that the heart rate responded more intensely to early morning exercise than to daytime exercise with a high load due, at least partly, to pronounced suppression of parasympathetic nerve activity. PMID- 12653195 TI - Non-linear dynamic analysis of hemodynamic parameters in an undulation type artificial heart system. AB - Undulation pump total artificial heart (UPTAH) is a unique total artificial heart implant (TAH) using an undulation pump that is a continuous blood flow pump. To evaluate the autonomic nerve function mediating the circulation system, we analyzed the hemodynamic parameters during animal experiments with UPTAH using the non-linear mathematical analyzing technique, including chaos and fractal theory. Adult female goats were used for the implantation of UPTAH. The natural heart was replaced with UPTAH under extra-corporal circulation. The conductance- and arterial pressure-based control method (1/R control) was applied on the 5th to 7th post-operative day as the influences of the cardiopulmonary bypass circulation were diagnosed to be terminated. Hemodynamic parameters were recorded on the data recorder, and non-linear mathematical analysis was performed. For the quantitative evaluation of the strange attractor, which was the characteristics of the deterministic chaos, the fractal dimension analysis was carried out. As a result, hemodynamic parameters fluctuated on the time axis and showed fractal characteristics, which were thought to be the characteristics of the deterministic chaos. The reconstructed attractor of the hemodynamics showed various behaviors according to changes in the situation of the goats. These results suggest that non-linear dynamical analysis might be useful in monitoring the circulatory regulatory system in artificial heart circulation. PMID- 12653196 TI - Non-linear dynamic analysis of hemodynamic behavior during virtual reality immersion. AB - Several years ago, a famous accident occurred in Japan. Hundreds of children, who were watching a cartoon on television, suddenly complained of spasms and vertigo, and were taken to hospital. In this study, the autonomic nervous system was evaluated during audiovisual stimulation with three dimensional Virtual Reality (VR) imaging. In our previous studies, we designed the diagnosis machine for an autonomic function using multi-parameters, including electrocardiography, arterial blood pressure, respiration and stroke volume as detected by ultrasonic cardiography. Healthy adult volunteers were used in this experiment with their satisfactory informed consent. The three-dimensional content for VR included dinosaur images in a pre-historic scene. The content was projected on a wide screen and volunteers watched an audiovisual screen for about 20 minutes and the 3-D and 2-D images were compared. There was no significant arrhythmia during experiments in both images. No significant alteration was observed in the quantified hemodynamic data during the experiment. Spectral analysis was performed to evaluate the heart rate variability (HRV) during the experiment. LF, HF and LF/HF of HRV were calculated. However, there were no significant changes during the experiment. Significant change was observed in the fractal dimension of the stroke volume during 2-D and 3-D image VR immersion. Our results suggest that a significant response was observed in the autonomic function according to the 2-D or 3-D images. Our study, which aims at safe audiovisual stimulating equipment, must be developed. PMID- 12653194 TI - Differences in sympathetic and vagal effects on paroxysmal atrial fibrillation: a simulation study. AB - The incidence of paroxysmal atrial fibrillation (AF) is affected by circadian variations in the vago-sympathetic balance. It is well known that both sympathetic and vagal effects increase the onset of paroxysmal AF, due to the shortened action potential duration. However, the reason why the vagally-mediated paroxysmal AF is maintained more than the adrenergically-mediated paroxysmal AF has remained unclear. In order to clarify this, we performed the following computer simulations. First, we constructed a homogeneous two-dimensional myocardial sheet (4.5 x 2.25 cm), using a bidomain ion channel model. The sympathetic and vagal effects were achieved by modifications of the ion channel conductance (Sympathetic effect: increased gSI and increased gK. Vagal effect: increased gK and increased gK1 with or without the dispersion of refractoriness). We found that the sympathetic effect shortened the action potential duration and flattened the restitution slope; therefore, this effect promoted spiral wave induction and restrained the spiral wave breakup. On the other hand, we found that the vagal effect also shortened the action potential duration and flattened the restitution slope; however, this effect promoted spiral wave breakup, due to the increase in both the IK1 and the dispersion of refractoriness. Overall, the differences between the sympathetic and vagal effects on the tendency toward spiral wave break-up may explain the reason why adrenergically-mediated paroxysmal AF terminates spontaneously and vagally-mediated paroxysmal AF tends to be maintained. In conclusion, our results may be helpful in understanding the difference in the action of sympathetic and vagal effects on paroxysmal AF. PMID- 12653197 TI - Non-linear dynamics applied to human respiratory movement during sleep. AB - We investigated the relationship between approximate entropy (ApEn) and correlation dimension (D2) in respiratory movement to reveal the non-linear dynamics in respiration during sleep. Seven healthy volunteers participated in this study. We recorded respiratory movement during sleep at night. We estimated the values of D2 and ApEn in respiratory movement. A significant relationship was observed between ApEn and D2 during sleep (r = 0.67, P < 0.001). Surrogate data analysis revealed that the respiratory movements had significant non-linear property. This evidence suggests that ApEn and D2 may become new indices to evaluate non-linear respiratory dynamics during sleep. PMID- 12653198 TI - Partial spectral element in the chronome of a human neonatal heart rate at term. AB - An initial infradian prominence characterizes the spectral element of the chronome or the time structure of half-hourly heart rates during the first 40 days after birth of a boy born at term. Immediately pertinent infradian prominence, which was previously documented for many premature infants and anticipated from the integration of 2-day segments of half-hourly data from infants born at term, is now displayed longitudinally. Dominating first are an about-weekly (circaseptan) or half-weekly (circasemiseptan) and an about 26-day (circatrigintan) component, with a subsequent change from infradian to circadian prominence. PMID- 12653199 TI - Chaos and chronos in the human brain: a tribute to Franz Halberg. AB - In order to understand the significance of brain signals, chaotic dynamics of the EEG were analyzed during parabolic flight. Chaotic and other measures of brain waves and eye movements change their characteristics as a function of a host of relatively slow components, such as an about-monthly cycle and early in human ontogeny, starting in gestation. Structural and functional redundancy, yet with diversity in space and time, may serve for adjustment to changes in the environment of the species and thus of the individual. With increasing age, with the accumulation of structured information, the order-to-noise ratio increases, suggesting that for a while during growth, the making of order out of chaos may define the course of development in time. PMID- 12653200 TI - The role of tetrahydrobiopterin in superoxide generation from eNOS: enzymology and physiological implications. AB - Tetrahydrobiopterin (BH4) is a ubiquitous pteridine metabolite that serves as a NOS cofactor. Recently, we showed that BH4 efficiently inhibits superoxide generation from the heme group at the oxygenase domain of eNOS. This role indicates that BH4 acts as a redox switch in the catalytic mechanism of the enzyme, which may have important consequences in the physiology of the endothelium. Here the mechanism by which BH4 inhibits superoxide release from eNOS and the "uncoupling" effects of oxidized BH4 metabolites are presented. The implications of the disparate actions of fully reduced and oxidized BH4 metabolites in the control of eNOS biochemistry are discussed in the light of clinical data indicating that BH4 levels are important in the regulation of superoxide levels and of endothelial reactivity. PMID- 12653201 TI - Hemin: a possible cause of oxidative stress in blood circulation of beta thalassemia/hemoglobin E disease. AB - A correlation between endogenous hemin and pro-oxidant activity was revealed in serum of beta-thalassemia/hemoglobin E disease (beta-thal/Hb E), which is the most common prevalent type of thalassemia in Thailand. The technique of low temperature electron spin resonance spectroscopy was used for characterization and quantification of high spin ferric heme, which had been identified as hemin (iron (III)-protoporphyrin IX). Hemin was present at levels ranging from 50 to 280 microM in serum of beta-thal/Hb E but not detectable in serum of non thalassemia. Pro-oxidant activity in serum of beta-thal/Hb E was demonstrated by luminol-mediated chemiluminescence, a sensitive method for screening of free radical generation in vitro. In the presence of H2O2, the chemiluminescence intensity (CL) was about 20 fold enhanced in serum of beta-thal/Hb E, indicating its extensive pro-oxidant activity. The CL showed a good correlation with serum heroin, r = 0.778 (p < 0.001), while the correlations with total serum iron and serum ferritin were 0.260 (p = 0.259) and 0.519 (p = 0.004), respectively. Our finding suggested that serum hemin readily catalyzed free radical reactions and it may contribute a major pro-oxidant in blood circulation of beta-thal/Hb E. PMID- 12653202 TI - Oxidative stress and potential free radical damage associated with photocopying. A role for ozone? AB - OBJECTIVE: To study the relationship between oxidative stress and potential free radical damage associated with photocopying and to explore a role for ozone emitted during the photocopying process. METHODS: 80 photocopying operators (PO) and 80 healthy volunteers (HV) were enrolled in a random control study design, in which the level of lipoperoxide (LPO, thiobarbituric acid reactive substances, TBARS) in erythrocytes and the levels of vitamin C (VC), vitamin E (VE) and beta carotene (beta-CAR) in plasma as well as the activities of superoxide dismutase (SOD) and catalase (CAT) in erythrocytes were determined by spectrophotometric methods. RESULTS: Compared with the average values of the above biochemical parameters in the HV group, the average value of LPO (TBARS) in erythrocytes in the PO group was significantly increased (P < 0.0001), while the average values of VC, VE and beta-CAR in plasma as well as those of SOD and CAT in erythrocytes in the PO group were significantly decreased (P < 0.0001). Pearson product-moment correlation analysis showed that with the increase of the ozone level in photocopying sites and the PO duration of exposure to ozone, the level of LPO in erythrocytes in the operators was increased (P < 0.001), while the levels of VC, VE and beta-CAR in plasma as well as the activities of SOD and CAT in erythrocytes in the operators were decreased (P < 0.01-0.0001). CONCLUSION: The findings in this study suggest that ozone causes oxidative damage in copier operatives. PMID- 12653203 TI - Synthesis, structure-activity relationship and in vitro evaluation of coelenterazine and coelenteramine derivatives as inhibitors of lipid peroxidation. AB - Coelenterazine (2-p-hydroxybenzyl-6-(3'-hydroxyphenyl)-8-benzyl-3,7 dihydroimidazolo[1,2-a]pyrazin-3-one, CLZn) and coelenteramine (2-amino-3-benzyl 5-(4'-hydroxyphenyl)-1,4-pyrazine, CLM), first described as luciferin and etioluciferin, respectively, of bioluminescent systems in marine organisms are endowed with antioxidant properties. This study was aimed at understanding the structural basis of their chain-breaking properties and at designing new compounds with improved antioxidative properties. For this, a series of 2-amino 1,4-pyrazine derivatives and their related imidazolopyrazinones were synthesised and examined for their capacity to inhibit lipid peroxidation in linoleate micelles subjected to the peroxidizing action of AAPH. Structure-activity relationship studies indicated that the reduction of the peroxidation rate by CLM is mainly determined by the concomitant presence of 5-p-hydroxyphenyl and 2-amino groups in para position. The lipophilic character of substituents also affected this effect. All imidazolopyrazinones induced a lag-time before the onset of the peroxidation process. The hetero-bicyclic imidazolopyrazinone moiety appears as the main contributor to this activity while phenol groups play little role in it. On the other hand, phenol groups were required for the reduction of the peroxidation rate after the lag-phase. The introduction of a supplementary p hydroxyphenyl substituent at C8 position did not increase chain-breaking properties. The substitution of the C5-p-hydroxyphenyl with a catechol moiety or the introduction of a second amino group on the pyrazine ring yielded the most active compounds, superior to imidazolopyrazinones and reference antioxidants like epigallocatechin gallate, vitamin E and trolox. The strong antioxidant properties of 2,6-diaminopyrazines are not dependent on the presence of hydroxyl groups indicating that their reaction mechanism differs from that of 2-amino-1,4 pyrazine derivatives. PMID- 12653204 TI - Direct detection of nitric oxide and its roles in maintaining gastric mucosal integrity following ethanol-induced injury in rats. AB - In gastric mucosal injury, nitric oxide (NO) plays both cytoprotective and cytotoxic roles, and the NO level is one determinant of these dual roles. We employed electron paramagnetic resonance (EPR)-spectrometry combined with an NO trapping technique to directly evaluate NO production in ethanol-induced gastric injury in rats. The rat stomach, mounted on an ex vivo chamber, was perfused with ethanol (12.5 and 43%), and NO levels in mucosal tissues were measured during perfusion. Luminal nitrite/nitrate (NOx) content, mucosal blood flow, area of mucosal injury, transmucosal potential difference (PD), and luminal pH were simultaneously monitored with/without preadministration of the NO synthase inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME). NO levels in the gastric tissue increased during ethanol perfusion, and luminal NOx levels increased after the perfusion, accompanying an increase in the area of mucosal injury and changes in physiological parameters. Preadministration of L-NAME aggravated the gastric mucosal damage and suppressed increases in mucosal blood flow in a dose-dependent manner. These results demonstrate that endogenous NO produced in ethanol-induced gastric injury contributes to maintenance of mucosal integrity via regulation of mucosal blood flow. PMID- 12653205 TI - Gas-phase disproportionation of nitric oxide at elevated pressures. AB - T.P. Melia's chemical kinetics study of the disproportionation of nitric oxide (NO), 3NO --> NO2 + N2O, (Melia, T.P. (1965) J. Inorg. Nucl. Chem., 27, 95-98), which is the most quoted quantitative investigation presently available, revealed a rather strong dependence of the effective rate constant, Kk' of Melia's third order rate law, -d[NO]/dt = Kk'[NO]3, on the initial pressure of NO. In order to estimate extent of accumulation of NO2 and N2O as a function of time by integration of the rate law, we have evaluated the dependence of the effective rate constant as a function of pressure and thus as a function of time on the basis of the non-ideality of NO gas. Although our approach is crude in that the non-idealities of NO2 and N2O and other NOx products and a probable deviation of the gas mixture from the Dalton's law have not been considered, it provides a means for approximately estimating the rate of accumulation of NO2 and N2O based on Melia's data. According to these calculations, the extent of the disproportionation is generally negligible at low initial pressures, e.g. 5 atm or less, while at 200 atm, the mole fractions of NO2 and N2O can become as high as 12-13% only after 10 days. These values are alarmingly high for handling pressured NO- in N2-mixture in either research or clinical settings. This information must be borne in mind when compressed NO in commercial cylinders is employed in high precision experiments. Disproportionation of NO under pressure also deserves attention in inhalation of low doses of NO in the treatment of diseases characterized by pulmonary hypertension and hypoxemia. PMID- 12653207 TI - MRP1-transfected cells do not show increased resistance against oxidative stress. AB - Sensitivity of V79 Chinese hamster cells and V79 cells transfected with human MRP1 gene to several agents inducing oxidative stress was compared. Cells overexpressing MRP1 did not show increased resistance to tert-butyl hydroperoxide, diamide, paraquat, menadione, dichromate and carmustine as estimated by cell survival and DNA damage assessed by comet assay. These findings suggest that overexpression of MRP1 does not confer increased resistance to oxidative stress. PMID- 12653206 TI - Xanthine oxidase mediates cytokine-induced, but not hormone-induced bone resorption. AB - Reactive oxygen species (ROS) such as hydrogen peroxide (H2O2) have been implicated as mediators of osteoclastic bone resorption. Xanthine oxidase (XO) a ubiquitous enzyme is widely known for its production of these ROS. We therefore evaluated the potential of XO as a source of ROS in cytokine-and hormone-induced bone resorption. XO activity in rat calvarial osteoblasts was found to be significantly elevated upon stimulation by the cytokines, TNFalpha and IL-1beta. These cytokines also caused a dose related increase in bone resorption of mouse calvariae, which was significantly inhibited by catalase (10 IU/ml). Allopurinol, the competitive inhibitor of XO, also caused a dose related (1-50 microM) inhibition of TNFalpha (20 ng/ml) and (0.01-10 microM) IL-1beta (50 IU/ml) induced bone resorption, respectively. PTH- and 1,25-(OH)2 Vitamin D3-induced bone resorption could also be inhibited by catalase (100 IU/ml) but was unaffected by allopurinol, indicating that another mediator, other than XO, is required for hormone-induced bone resorption. These results demonstrate, that modulation of the redox balance in the bone microenvironment, which contains XO, can affect the bone resorbing process. Therefore, XO may play a pivotal role in cytokine-induced bone resorption and, if manipulated appropriately, could show a therapeutic benefit in inflammatory bone disorders such as RA. PMID- 12653208 TI - Enhancement of glucose transport in rat thymocytes by different radical sources. AB - This study demonstrates that oxidative stress induced in rat thymocytes by the hydrophilic 2,2'-azobis(2-amidinopropane)dihydrochloride (AAPH), the lipophilic cumene hydroperoxide (CumOOH) and the freely diffusible H2O2 is associated with an activation of facilitative glucose transport rate, mediated by GLUT1, the major transporter in this cell type. We compared the effects of the three tested radical sources on the kinetic transport parameters, showing that the transport rate enhancement in the treated cells can be ascribed to an increase in the Vmax value, apart from the site of generation of the oxidative stress. The enhancement of glucose transport by the three oxidants in thymocytes was significantly attenuated both by protein tyrosine kinase inhibitors as genistein and tyrphostin A23 and by U73122, a phospholipase C inhibitor. Genistein and U73122 reversed also the cited increase of Vmax values. It is concluded that the stimulation of glucose transport in response to different oxidants is mediated, at least in part, through reactive oxygen species (ROS)-induced stimulation of protein tyrosine kinase and phospholipase C pathways. PMID- 12653209 TI - Identification of the binding site of methylglyoxal on glutathione peroxidase: methylglyoxal inhibits glutathione peroxidase activity via binding to glutathione binding sites Arg 184 and 185. AB - Methylglyoxal (MG), a physiological alpha-dicarbonyl compound is derived from glycolytic intermediates and produced during the Maillard reaction. The Maillard reaction, a non-enzymatic reaction of ketones and aldehydes with amino group of proteins, contributes to the aging of proteins and to complications associated with diabetes. In our previous studies (Che, et al. (1997) "Selective induction of heparin-binding epidermal growth factor-like growth factor by MG and 3 deoxyglucosone in rat aortic smooth muscle cells. The involvement of reactive oxygen species formation and a possible implication for atherogenesis in diabetes". J. Biol. Chem., 272, 18453-18459), we reported that MG elevates intracellular peroxide levels, but the mechanisms for this remain unclear. Here, we report that MG inactivates bovine glutathione peroxidase (GPx), a major antioxidant enzyme, in a dose- and time-dependent manner. The use of BIAM labeling, it was showed that the selenocysteine residue in the active site was intact when GPx was incubated with MG. MALDI-TOF-MS (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry) and protein sequencing examined the possibility that MG modifies arginine residues in GPx. The results show that Arg 184 and Arg 185, located in the glutathione binding site of GPx was irreversively modified by treatment with MG. Reactive dicarbonyl compounds such as 3-deoxyglucosone, glyoxal and phenylglyoxal also inactivated GPx, although the rates for this inactivation varied widely. These data suggest that dicarbonyl compounds are able to directly inactivate GPx, resulting in an increase in intracellular peroxides which are responsible for oxidative cellular damage. PMID- 12653210 TI - The impact of different antioxidant agents alone or in combination on reactive oxygen species, antioxidant enzymes and cytokines in a series of advanced cancer patients at different sites: correlation with disease progression. AB - In the present study we tested the ability of different antioxidant agents, used alone or in combination, to reduce the reactive oxygen species (ROS) levels and to increase the glutathione peroxidase (GPx) activity. Moreover, we tested the ability of such antioxidant agents to reduce the serum levels of proinflammatory cytokines IL-6 and TNFalpha. Fifty-six advanced stage cancer patients with tumors at different sites were included in the study: they were mainly stage III (12.5%) and stage IV (82.1%). The study was divided into two phases. In the 1st phase 28 patients were divided into five groups and a single different antioxidant agent was administered to each group. The selected antioxidant agents were: alpha lipoic acid or carboxycysteine-lysine salt, amifostine, reduced glutathione, vitamin A plus vitamin E plus Vitamin C. In the 2nd phase of the study 28 patients were divided into five groups and a combination of two different antioxidant agents was administered to each group. The antioxidant treatment was administered for 10 consecutive days. The patients were studied at baseline and after antioxidant treatment. Our results show that all single antioxidants tested were effective in reducing the ROS levels and three of them in increasing GPx activity, too. Among the combinations of antioxidant agents, three were effective in reducing ROS, while three were effective in increasing GPx activity (arm 4 was effective in both instances). Comprehensively, the "antioxidant treatment" was found to be effective both on ROS levels and GPx activity. Moreover, the antioxidant treatment was able to reduce serum levels of IL-6 and TNFalpha. Furthermore, a correlation was shown between the Eastern Cooperative Oncology Group Performance Status of patients and blood levels of ROS, GPx activity, serum levels of proinflammatory cytokines. PMID- 12653211 TI - Methodologies for the sensitive and specific measurement of S-nitrosothiols, iron nitrosyls, and nitrite in biological samples. PMID- 12653212 TI - Oxidative stress is evident in erythrocytes as well as plasma in patients undergoing heart surgery involving cardiopulmonary bypass. AB - OBJECTIVE: The aim of this study was to analyse the level and progression of oxidative stress, in both plasma and erythrocytes, during heart surgery involving cardiopulmonary bypass. MATERIALS AND METHODS: Twenty two patients undergoing cardiac surgery and considered to present a high/severe level of surgical risk were selected. We took five blood samples at different times during the cardiac surgery and analysed TBARS, alpha-tocopherol, coenzyme Q and retinol in plasma and TBARS (baseline levels and induced by Fe2+-ascorbate oxidation), alpha tocopherol, coenzyme Q and catalase, superoxide dismutase and gluthatione peroxidase activity in erythrocytes. RESULTS: Plasma results shown a decrease in both alpha-tocopherol and retinol concentration after starting CPB with respect to the reference level (13.6 +/- 1.5 nmol ml(-1) vs. 22.0 +/- 3.0 nmol ml(-1) and 1.2 +/- 0.1nmol ml(-1) vs. 1.8 +/- 0.2 nmol ml(-1), respectively (p < 0.05)). In comparison, in erythrocytes, all antioxidants, both enzymatic and non-enzymatic, increased in activity or concentration after starting CPB. Erythrocyte TBARS, both baseline levels and induced levels, followed a similar pattern, with an increase after starting CPB with respect to the reference level (3.9 +/- 0.6 nmol mg(-1) of protein vs. 2.3 +/- 0.2 nmol mg(-1) of protein and 10.6 +/- 0.8 nmol mg(-1) of protein vs. 6.7+/- 0.6 nmol mg(-1) of protein, respectively (p < 0.05)). CONCLUSION: These results reveal an increase in oxidative stress after CPB, both in plasma and erythrocytes, and although the organism is capable of attenuating this stress by means of various antioxidative defence mechanisms, there is an increased possibility of post-CPB complications and thus of mortality. PMID- 12653213 TI - Immune cells redox state from mice with endotoxin-induced oxidative stress. Involvement of NF-kappaB. AB - The immune cells, such as phagocytes and lymphocytes, which use reactive oxygen species (ROS) for carrying out many of their functions, need appropriate levels of intracellular antioxidants to avoid the harmful effect of oxidative stress. In previous studies, we have observed changes in several functions of those leukocytes from female BALB/c mice with lethal endotoxic shock caused by intraperitoneal injection of Escherichia coli 055:B5 lipopolysaccharide (LPS) (100 mg/kg), which were associated with high ROS production. In the present study, we have investigated the redox state of the above mentioned immune cells in that lethal endotoxic shock model measuring the oxidant/antioxidant balance through the following parameters: production of ROS, proinflammatory cytokine TNFalpha reduced glutathione (GSH), oxidized glutathione (GSSG), superoxide dismutase (SOD) and catalase (CAT) activities, malonaldehyde (MDA) and transcription factor NF-kappaB expression at different times after LPS injection. The results show an increase in ROS, TNFalpha and MDA production in both cell types, being higher in macrophages than in lymphocytes. GSSG/GSH ratio was increased in both macrophages and lymphocytes after LPS injection. With respect to the activity of the antioxidant enzymes SOD and CAT were decreased in both macrophages and lymphocytes. The activation of the transcription factor NF-kappaB was stimulated in macrophages and lymphocytes. These results point out that both lymphocytes and macrophages, which are able to play an important role in host response to endotoxin, show an oxidative stress thus contributing to the pathogenesis of this septic shock. PMID- 12653214 TI - NO synthesis in human saliva. AB - Human saliva contains nitrate that is converted into nitrite by the activity of facultative, anaerobic bacteria of the oral cavity. Nitrite can be reduced to NO in the acidic gastric milieu; some NO may also form in the mouth at acidic pH values. In this paper, we show that bacteria (S. salivarius, S. mitis and S. bovis) isolated from saliva, may contribute to NO production in human saliva. NO formation by bacteria occurs at neutral pH values and may contribute to the antibacterial activity of saliva. PMID- 12653216 TI - Purity of different preparations of sodium 3,5-dibromo-4-nitrosobenzenesulphonate and their applicability for EPR spin trapping. AB - During the preparation of sodium 3,5-dibromo-4-nitrosobenzenesulphonate (DBNBS) of high purity for electron paramagnetic resonance (EPR) spin-trapping purposes, it was found that the material synthesised as part of the present study differed significantly from some commercially available samples of DBNBS. A thorough chemical characterisation of the contents of the various samples led to the conclusion that the preparations synthesised in the present study, as well as one of four commercially available samples, contained essentially pure DBNBS and had efficient spin-trapping activity. In contrast, the remaining three commercially available samples contained almost exclusively sodium 3,5-dibromo-4 nitrobenzenesulphonate, i.e. a one-oxygen oxidation product of DBNBS, and had little spin-trapping activity. The two compounds were readily separated by reverse-phase high performance liquid chromatography (HPLC). It was further found that the quality of DBNBS preparations may be determined by NMR spectrometry, IR spectrometry, fast atom bombardment-mass spectrometry (FAB-MS) and EPR spectrometry. In particular, UV-Visible spectroscopy may be used to determine A308/A280, which should be greater than 1.8 for a high purity DBNBS preparation. PMID- 12653215 TI - Preventing superoxide formation in epineurial arterioles of the sciatic nerve from diabetic rats restores endothelium-dependent vasodilation. AB - We have previously reported that in streptozotocin-induced diabetic rats that increased formation of superoxide and peroxynitrite is associated with impairment in vascular relaxation in epineurial arterioles of the sciatic nerve. In this study we demonstrate that pretreating epineurial arterioles from diabetic rats in vitro with alpha-lipoic acid, dihydrolipoic acid, tempol or arginine restores acetylcholine-mediated vascular relaxation to near the reactivity observed in vessels from control rats. Suggesting that increased oxidative stress and reduction in nitric oxide availability is partially responsible for the impairment in endothelium-dependent vasodilation observed in epineurial arterioles from diabetic rats. In contrast, pretreating epineurial arterioles from diabetic rats with aminoguanidine or allopurinol had no effect. Studies designed to investigate the source of superoxide formation provided results suggesting that complex I of the mitochondrial electron transport chain and NAD(P)H oxidase are responsible for the increase in superoxide formation observed with epineurial arterioles from the sciatic nerve. Pretreating epineurial arterioles from diabetic rats with the protein kinase C inhibitor bisindolymaleimide I (GF 109203X) improved acetylcholine-mediated vascular relaxation but did not prevent the increase in superoxide formation suggesting that activation of protein kinase C by oxidative stress is downstream of superoxide formation. These studies imply that increased superoxide formation via the mitochondrial electron transport chain and perhaps NAD(P)H oxidase is partially responsible for reduced vascular reactivity observed in epineurial arterioles of the sciatic nerve from diabetic rats. PMID- 12653218 TI - Effects of a direct injection of liposoluble iron into rat striatum. Importance of the rate of iron delivery to cells. AB - For a better understanding of the role of iron imbalance in neuropathology, a liposoluble iron complex (ferric hydroxyquinoline, FHQ) was injected into striatum of rats. The effects of two modalities of iron injections on brain damage, hydroxyl radical (*OH) production (assessed by the salicylate method coupled to microdialysis) and tissue reactive iron level (evaluated ex vivo by the propensity of the injected structure for lipid peroxidation) were examined. Rapid injection of FHQ (10 nmoles of 5 mM FHQ pH 3 solution over 1-min period) but not that of corresponding vehicle led to extensive damage associated with increased tissue free iron level in the injected region. Conversely, neither lesion nor free iron accumulation was observed after slow FHQ injection (10 nmoles of a 100 microM FHQ pH 7 solution over 1-h period) as compared to corresponding vehicle injection. Production of *OH was induced by slow FHQ injection but not by rapid FHQ injection, probably as a result of in vivo abolition of iron-induced *OH formation by acid pH. Indeed, rapid injection of FAC pH 7 (ferric ammonium citrate, 5 mM in saline) was associated with *OH formation whereas rapid injection of FAC pH 3 did not. Our results identify the rate of iron delivery to cells as an important determinant of iron toxicity and do not support a major role for extracellular *OH in damage associated with intracerebral iron injection. PMID- 12653217 TI - Iron release in erythrocytes and plasma non protein-bound iron in hypoxic and non hypoxic newborns. AB - Iron is released in a desferrioxamine (DFO)-chelatable form (DCI) when erythrocytes are challenged by an oxidative stress. In beta-thalassemic erythrocytes, both DCI content and release (after aerobic incubation for 24h) are increased and correlated with the fetal hemoglobin (HbF) levels. Since erythrocytes from newborns have an extremely high content of HbF and are exposed to conditions of oxidative stress, the release of iron in these erythrocytes was investigated. The erythrocyte DCI content was increased in preterm but not in term newborns as compared to adults, while the release was increased in both preterm and term erythrocytes. The level of plasma non protein-bound iron (NPBI), which was not detectable in adults, was much higher in preterm than in term newborns. When term plus preterm newborns were divided in two groups, normoxic and hypoxic, according to cord blood pH, it was found that both iron release and NBPI were markedly higher in the hypoxic newborns compared to normoxic ones. Similar results were also obtained when the preterm and term infants were considered separately on the basis of cord blood pH. Therefore, iron release and NPBI are higher when conditions of hypoxia occur. In fact, when the values for iron release and NPBI were separately plotted against cord blood pH values, significant negative correlations were seen in both cases. NPBI was correlated with iron release seen in all the newborns and a significant part of the released iron could be recovered into the incubation medium at the end of the incubation. PMID- 12653219 TI - Oxidative DNA damage induced by toluene is involved in its male reproductive toxicity. AB - Toluene is widely used as an organic solvent in various industries and commercial products. Recent investigations have shown that toluene may induce male reproductive dysfunctions and carcinogenicity. To clarify whether the toxicity results from the interference of endocrine systems or direct damage to reproductive organs, we examined the effects of toluene on the male reproductive system in rats, comparing to those of diethylstilbestrol (DES), a potent synthetic estrogen. Toluene (50, 500 mg/kg) or DES (2 mg/kg) injected subcutaneously to male Sprague-Dawley rats once a day for 10 days decreased the epididymal sperm counts and the serum concentrations of testosterone. The mRNA level for gonadotropin-releasing hormone receptor in the pituitary was decreased by DES, but not by toluene. On the contrary, 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) formation in testes, the biological marker for oxidative DNA damage, was increased by toluene but not by DES. These results suggest that toluene induces reproductive toxicity via direct oxidative damage of spermatozoa, whereas DES affects endocrine systems via the hypothalamo-pituitary-gonadal axis. Morphological findings supported the idea. To determine the mechanism of 8-oxodG formation in vivo, we examined DNA damage induced by toluene metabolic products in vitro. Minor toluene metabolites, methylhydroquinone and methylcatechols, induced oxidative DNA damage, and the methylcatechols induced NADH-mediated 8 oxodG formation more efficiently than methylhydroquinone did. We propose that oxidative DNA damage in the testis plays a role in reproductive toxicity induced by toluene. PMID- 12653220 TI - PON1 paraoxonase activity is reduced during HDL oxidation and is an indicator of HDL antioxidant capacity. AB - The aim of this study was to investigate the effect of HDL oxidation on PON1 paraoxonase activity. Also, we were interested in investigating the mechanism by which PON1 could be inactivated and the correlation between its enzymatic activity and the antioxidant properties of HDL. Three different oxidation systems were used for the HDL oxidation: (1) oxidation induced by THP1 cells, (2) oxidation induced by copper ions at a concentration 10 microM, and (3) oxidation induced by *OH and O2.- oxygen free radicals produced by gamma-radiolysis. HDL oxidation was followed by the measurement of lipid peroxide formation, and PON1 activity was determined by measuring the rate of paraoxon hydrolysis. Our results show that HDL oxidation is accompanied by a reduction in the PON1 paraoxonase activity. The extent of PON1 inactivation depends both on the extent of HDL oxidation and on the oxidation system used. The rates of HDL oxidation and PON1 inactivation were significantly correlated (r = 0.93, p < 0.0054). Our results show that oxidized HDL loses its protective effect toward LDL oxidation. The antioxidant action of HDL towards LDL oxidation and the degradation of PON1 paraoxonase activity were significantly correlated (r = 0.95, p < 0.04). PMID- 12653221 TI - Dietary nitrate inhibits stress-induced gastric mucosal injury in the rat. AB - Dietary nitrate is reduced to nitrite by some oral bacteria and the resulting nitrite is converted to nitric oxide (NO) in acidic gastric juice. The aim of this study is to elucidate the pathophysiological role of dietary nitrate in the stomach. Intragastric administration of nitrate rapidly increased nitrate and NO in plasma and the gastric headspace, respectively. Water-immersion-restraint stress (WIRS) increased myeloperoxidase (MPO) activity in gastric mucosa and induced hemorrhagic erosions by a nitrate-inhibitable mechanism. In animals that had received either cardiac ligation or oral treatment with povidone-iodine, a potent bactericidal agent, administration of nitrate failed to increase gastric levels of NO and to inhibit WIRS-induced mucosal injury. WIRS decreased gastric mucosal blood flow by a mechanism which was inhibited by administration of nitrate. These data suggested that the enterosalivary cycle of nitrate and related metabolites consisted of gastrointestinal absorption and salivary secretion of nitrate, its conversion to nitrite by oral bacteria and then to NO in the stomach might play important roles in the protection of gastric mucosa from hazardous stress. PMID- 12653222 TI - The antioxidant cocktail, effective microorganism X (EM-X), protects retinal neurons in rats against N-methyl-D-aspartate excitotoxicity in vivo. AB - Injection of the glutamate agonist N-methyl-D-aspartate (NMDA) into the vitreous body of rats resulted in severe degeneration of neurons in the retina, with a loss of 81% of ganglion cells and 43% of non-ganglion cells. The cocktail EM-X is a novel antioxidant drink derived from ferment of unpolished rice, papaya and sea weeds with effective microorganisms (EM-X). In animals treated with an intraperitoneal injection of EM-X, the loss of ganglion cells was reduced to 55% and that of non-ganglion cells to 34% when compared to untreated NMDA-injected retinas. Cell degeneration resulting from NMDA excitotoxicity, is thought to be mediated via oxidative stress mechanisms. The neuroprotective effect of the EM-X in this system is therefore likely to be due, at least in part, to its flavonoids, saponins, vitamin E and ascorbic content. PMID- 12653223 TI - Nitric oxide exposure of CC531 rat colon carcinoma cells induces gamma glutamyltransferase which may counteract glutathione depletion and cell death. AB - Gamma-glutamyltransferase (GGT) has a central role in glutathione homeostasis by initiating the breakdown of extracellular GSH. We investigated in the present study whether nitric oxide exposure of CC531 rat colon carcinoma cells modulates GGT and how the activity of the enzyme affects the level of intracellular GSH. The data show that GGT activity was induced in a dose-related manner by two NO donors (spermineNONOate and nitrosoglutathione) and that antioxidants partly inhibited the induction. SpermineNONOate lowered intracellular GSH and induced apoptosis. Cultivating the cells in cystine-depleted medium also resulted in a 50% lowering of GSH, but this was avoided when GSH was added to the medium. This effect was mediated by the activity of GGT and shown after inhibiting GGT activity with acivicin and cyst(e)ine transporters with alanine and homocysteic acid. This shows that the cells benefit from GGT in maintaining the intracellular GSH level. Cells with induced GGT activity obtained after NO incubation showed a higher uptake rate of cysteine (2-fold), measured by incubating the cells with 5S radiolabeled GSH. The enzyme was also induced by interferon-gamma and tumor necrosis factor-alpha, but this induction was not connected to activation of the endogenous nitric oxide synthase, as the addition of aminoguanidine, a NO synthase inhibitor, did not affect the induction. The present study shows that the activity of GGT is upregulated by NO-donors and that the colon carcinoma cells, when cultivated in cystine-depleted medium, benefit from the enzyme in maintaining the intracellular level of GSH. Thus, the enzyme will add to the protective measures of the tumor cells during nitrosative stress. PMID- 12653224 TI - Electrochemical and EPR characterization of 1,4-dihydropyridines. Reactivity towards alkyl radicals. AB - This work reports the electrochemical oxidation of a series of three synthesized 4-substituted-1,4-dihydropyridine derivatives in different electrolytic media. Also, an EPR characterization of intermediates and the reactivity of derivatives towards ABAP-derived alkyl radicals are reported. Dynamic, differential pulse and cyclic voltammetry studies on a glassy carbon electrode showed an irreversible single-peak due to the oxidation of the 1,4-dihydropyridine (1,4-DHP) ring via 2 electrons to the corresponding pyridine derivative. Levich plots were linear in different media, indicating that the oxidation process is diffusion-controlled. Calculated diffusion coefficients did not exhibit significant differences between the derivatives in the same medium. The oxidation mechanism follows the general pathway (electron, H+, electron, H+) with formation of an unstable pyridinium radical. One-electron oxidation intermediate was confirmed with controlled potential electrolysis (CPE) and EPR experiments. On applying N-tert-butyl-alpha phenylnitrone (PBN) and 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as the spin trap, these unstable radical intermediates from the oxidation of 1,4-DHP derivatives were intercepted. The final product of the CPE, i.e. pyridine derivative, was identified by GC-MS technique. Direct reactivity of the synthesized compounds towards alkyl radicals was demonstrated by UV-Vis. spectroscopy and GC-MS technique. Results indicate that these derivatives significantly react with the radicals, even compared with a well-known antioxidant drug such as nisoldipine. PMID- 12653225 TI - Assessment of the value of anorganic bone additives in sinus floor augmentation: a review of clinical reports. AB - In order to objectively assess the value of anorganic bone additives in sinus floor augmentation, a review of the English literature till May 2002 was carried out. The studies or parts of studies had to include at the least two patients, with a follow-up of at least 3 months, whilst histomorphometric data ought to be available of biopsies taken from the core of the graft. Only 12 studies fulfilled these criteria. Firm conclusions could not be drawn because there were substantial differences in histomorphometric techniques, healing periods and ratio autogenous bone anorganic additives, but it seemed that autogenous bone without anorganic additives resulted in the highest amount of bone after a 4-6 months healing period, while hydroxyapatite and bovine bone mineral, used as a bone substitute, gave the lowest amount of bone. No correlation between bone volume and time of graft healing could be found. The consequences, however, for implant placement and survival are at present, not fully understood. A plea is made for systematic prospective clinical and experimental studies to assess the various bone additives and ratios of autogenous bone to bone additives in composite grafts analysed with an adequate histomorphometric technique. Part of this study was presented at the 15th ICOMS, Durban, South Africa, 19-24 May 2001. PMID- 12653226 TI - Image-based planning and clinical validation of zygoma and pterygoid implant placement in patients with severe bone atrophy using customized drill guides. Preliminary results from a prospective clinical follow-up study. AB - The zygoma implant has been designed for those situations where there is insufficient bone in the upper jaw, which would otherwise require onlay or inlay (sinus) bonegrafts. The aim of the study was to present and validate a planning system for implant insertion based on preoperative CT imaging. It allows the surgeon to determine the desired position of different kinds of implants. Finally a customized drill guide is produced by stereolithography. In this study, zygoma, pterygoid and regular platform implants were used. The treatment protocol is validated through 12 case studies, selected at random from the total patient group (n=29 patients). From postoperative images, the exact implant location is determined and the deviation of axes between planned and inserted implants is calculated. In this in vivo study, displacements, varying according to the type of implant and the location of the implants, were observed. From a clinical standpoint, most of the inserted implants were judged to be adequately sited. A prospective clinical follow-up study was performed on all 29 patients. Although all patients presented with severe maxillary atrophy, excellent cumulative survival rates (92%) for the zygoma implants and 93% for regular platform implants have been obtained. PMID- 12653227 TI - An evaluation of clinical and electrophysiologic tests in nerve injury diagnosis after mandibular sagittal split osteotomy. AB - The yield of clinical sensory tests and electrophysiologic tests in the diagnostics of inferior alveolar nerve (IAN) damage after bilateral sagittal split osteotomy (BSSO) was studied. The diagnostic value of these tests was evaluated by comparing the test results to the degree of nerve damage at the end of the operation as documented by means of the intraoperative nerve conduction recording of the IAN. Twenty patients undergoing BSSO were analysed preoperatively and 2 weeks postoperatively. The frequency of the IAN disturbance ranged from 10% to 94% depending on the test method and the test site used. Of the clinical sensory tests, the touch detection threshold (TD) test was the most sensitive and clinically useful test. It also correlated best with the electrophysiologically verified intraoperative nerve damage (R = -0.603, P = 0.017 on the right, R = -0.626, P = 0.01 on the left). The blink reflex and quantitative cold detection threshold tests were almost as often abnormal as the TD-test, but nerve conduction study (NCS) was the most sensitive (88%) of all clinical and electrophysiologic tests. The frequency of abnormal findings in the electrophysiologic tests indicating IAN injury, 75% on the right side and 90% on the left side, corresponded exactly with the figures of subjective sensory alteration. Almost all electrophysiologic tests showed obvious associations with the objectively verified IAN damage. All tests, except the NCS, showed only moderate sensitivity. Specificity of the tests was generally high, the only exceptions being the TD test and the NCS. To increase the diagnostic accuracy of the testing and to detect different types of damage in different nerve fibre populations, a combination of different sensory and electrophysiologic tests is recommended. PMID- 12653228 TI - Temporomandibular joint ankylosis: report of 14 cases. AB - The authors present a review of 14 patients with temporomandibular joint ankylosis treated between March 1992 and February 1997. Etiology of the ankylosis was trauma in four patients, ear infection in two, systemic infection in one case, congenital in another, and unknown in six. Patients were divided into two groups, according to their age: 16 years and under and over 16 years of age. The basic principle of surgical treatment in both groups is ample access for osseous resection and coronoidectomy. Costochondral grafts were used in group one (nine patients), while interposition of a silicone block, was performed in the second group (five patients). Follow-up evaluations were from twelve to 53 months (average 28.2 months). One case of recurrence occurred in the first group and no recurrences in the second group. The average long-term mouth opening in both groups was 32.8 mm. PMID- 12653229 TI - The prognostic implications of the surgical margin in oral squamous cell carcinoma. AB - The prime objective of tumour ablation in oral squamous cell carcinoma (OSCC) is the removal, with a 'margin' of normal tissue, of the whole tumour. Definition of what constitutes margin involvement varies. This study aims to examine the factors associated with close and involved surgical margins in the management of OSCC. A cohort of 200 consecutive patients with previously untreated OSCC provided the material for the study. Various clinical, operative and pathological parameters were related to the status of the surgical margin, as well as time to recurrence, and survival. Cox regression analysis of the survival was also undertaken. Of the 200 patients 107 (53.5%) had clear margins, 84 (42%) close and 9 (4.5%) involved. Poor correlation was found between the status of the surgical margin and clinical factors, but in contrast high correlation between histological indicators of aggressive disease and close or involved surgical margins. These results imply that close surgical margins in OSCC could be regarded as an indictor of aggressive disease. PMID- 12653230 TI - Influence of radiation therapy on reconstructive flaps after radical resection of head and neck cancer. AB - This study was undertaken to evaluate the influence of radiotherapy (RT) on reconstructive flaps after radical excision of head and neck cancer. Eighty-eight reconstructive flaps were used in 82 patients who received RT either before (PREOP group, 14 flaps) or after (POSTOP group, 74 flaps) surgery. The success and healing rates of the flaps were evaluated. The success and healing rates were lower in the PREOP group than in the POSTOP group (86% vs 99%, P=0.026 for success; and 64% vs 95%, P=0.003 for healing). The rate of acute radiation reaction in flaps in the POSTOP group was significantly lower than in the surrounding normal tissues (35% vs 84%, P=0.003). Late side effects of RT were rare. Our results suggest good radiation tolerance of reconstructive flaps after radical tumour excision in the head and neck region. Success and healing of the flap are likely to be better if RT is administered after surgery. PMID- 12653231 TI - The diagnostic value of ultrasonography to detect occult lymph node involvement at different levels in patients with squamous cell carcinoma in the maxillofacial region. AB - The purpose of this study was to evaluate ultrasonography (US) and computed tomography (CT) in detecting lymphnodes of the neck affected with squamous cell carcinoma. From 1987 to 1999 the data from 203 untreated patients with a diagnosis of cancer in the maxillofacial have been investigated. Of these, 115 had a primary squamous-cell carcinoma. US diagnosis was made by an oral- and maxillofacial surgeon experienced in US of the head and neck. CT diagnosis was made by a well-experienced radiologist. The following lymph node levels were assesed: level I (submental and submandibular lymphnodes), level II (lymphnodes distal to level I and confined to the region above the skin crease at or just below the level of the thyroid notch), level III (lymphnodes distal to level II and confined to the anterior cervical triangle including those deep to the sternocleidomastoid muscle), and level IV (lymphnodes distal to level III and confined to the posterior cervical triangle). For all levels US yielded a sensitivity of 71%, and a specificity of 87%, while CT showed a sensitivity of 32% and a specificity of 96%. The sensitivity of US decreased from level I to level IV, whereas the specificity increased from level I to level IV. For lymphnode levels I and II US may be useful for the detection of local metastases while for the other levels the application of advanced techniques of US may have to be investigated. PMID- 12653232 TI - Soft tissue sarcoma of the head and neck region in adults. AB - Soft tissue sarcomas (STS) are rare solid tumours accounting for less than 1% of all malignancies and are very unusual in the head and neck region. Histopathologically diagnosed cases of STS treated at Regional Cancer Centre (RCC). Trivandrum, India, between January 1989 and November 2000 were analyzed retrospectively. Survival analysis was carried out by Kaplan-Meier method and curves were compared using log rank test. A total of 22 cases were seen during the study period. The mean age of the patients was 36.4 years with male-to-female ratio of 2:1. The neck, lower jaw, tongue, cheek, scalp and maxilla were the commonest sites affected. None of the patients had palpable neck nodes or distant metastasis at presentation. All the patients were treated with primary surgical resection and this was followed by adjuvant treatment in 14 cases (63.6%). After a median follow-up of 14.5 months, two patients died, six developed local recurrence, four developed metastatic disease and another patient developed second primary sarcoma. The overall 5-year survival was 80% while the 5-year disease-free survival rate was 24.1%. The majority of the patients failed within first and second year. None of the parameters except grade of tumour (P=0.008) were found to have a significant effect on survival. The overall survival rate for patients with soft tissue sarcoma of the head and neck is good, however, disease-free survival is poor as the majority of these fail within 2 years of completing treatment. PMID- 12653233 TI - Lipomas of the oral cavity: clinical findings, histological classification and proliferative activity of 46 cases. AB - Lipomas represent about 1 to 5% of all neoplasms of the oral cavity. Although relatively common, few large series of intraoral lipomas and its variants are seen in the literature. Therefore, the authors present the clinical, histological and immunohistochemical features of 46 cases of intraoral lipomas reviewed from the files of the University of Campinas Dental School from 1970 to 2001. Most of the cases affected adults, without gender predilection, and the main involved sites were the buccal mucosa (21 cases), tongue (six cases), lips (six cases) and floor of mouth (five cases). The histological analysis revealed 21 cases of lipoma, 18 fibrolipomas, four intramuscular lipomas, two minor salivary gland lipomas and one spindle cell lipoma. PCNA and ki-67 expression indexes were higher in spindle cell lipoma, intramuscular lipomas and fibrolipomas compared to common lipomas, but the differences were not statistically significant. All lesions were removed surgically and none showed recurrence, regardless of the various proliferative activities. PMID- 12653235 TI - Cross-sectional characteristics of the masseter muscle: an ultrasonographic study. AB - Cross-sections of the masseter muscle may now be measured non-invasively using ultrasonography. The purpose of the present study was to determine (1) whether the scanning level may affect cross-sectional measurements and (2) whether measurements made at identical levels may be reproducible. The study included 42 asymptomatic volunteers. Unilateral ultrasonographic investigation was performed with a linear (B-scan) 7.5 Mhz small-part transducer to register cross-sections of the masseter muscle on five different levels. Scans were made on the relaxed and contracted muscle. Measurements were made in two sessions with a time interval of at least 5 min. Statistical analysis consisted of univariate analysis of variance for repeated measurements. Data were analysed for reproducibility by using the method error and measurement error. For the ultrsonographic measurements the main effects of the variables 'session' (P=0.0001), 'level' (P=0.0001), and 'condition' (P=0.0001) were significant. Analysis of simple session-within-level effects revealed a significant difference between the repeated measurements for the most upper level (P=0.022), the upper level (P=0.012), and the most lower level (P=0.0001) of the relaxed muscle. An additional significant effect was found for the most lower level of the contracted muscle (P=0.015). Cross-sections evaluated at the middle (method error=0.31 mm; measurement error=2.0%) and lower level (method error=0.32 mm; measurement error=2.4%) of the contracted muscle were the most reproducible. The conclusion is that ultrasonography is a reproducible method for measuring cross sections at the middle and lower level of the contracted masseter muscle. PMID- 12653234 TI - Effects of latency period in a rabbit mandibular distraction osteogenesis. AB - Recently, distraction osteogenesis has been widely used in the craniomaxillofacial skeleton. There are varying views, however, about the need for a latency period for mandibular distraction, which has been reported from 0 to 14 days. We used a rabbit model to determine the ideal latency period for mandibular distraction osteogenesis. We performed a subperiosteal corticotomy of the bilateral mandible and then applied an external device. We used 0-, 2-, 5-, and 10-day latency periods. Distraction was done at a rate of 0.5 mm per day (0.25 mm every 12 h) to elongate by 5 mm after each latency period. Hematoxylin eosin stain was used for histologic observation, and bone labelling was observed with a fluorescence microscope. In the 0-day latency, the lengthened region was filled with fibrous tissue. Cartilage formed like isles along the lengthened edge. In the 10-day latency, the lengthened region was almost completely filled with woven bone. In the 2- and 5-day latency, there were mineralization zones at the periphery of this fibrous interzone. The primary osteon remodelling zones were located at both ends of the zones of mineralization zones. In the 5-day latency, new bone formation was observed around the periosteum. PMID- 12653236 TI - Morphological changes of parotid glands following adjuvant arthritis and ibuprofen treatment in rats. AB - Drug administration and numerous systemic diseases may cause morphological changes of the parotid gland. The aim of this study was to investigate the possible relationship between experimental adjuvant arthritis following ibuprofen treatment and morphological alterations of the parotid glands in rats. Freud's adjuvant was injected intradermally into the plantar surface of the hind paw of the animals to induce experimental arthritis. Ibuprofen was administrated per os (17 mg/kg/day). Both adrenals and parotid glands were isolated and their absolute and relative weights were evaluated. A full histological examination of parotid glands took place. The diameter of the foot as well as the serum levels of rheumatoid factor was measured. In conclusion, both experimental adjuvant arthritis and ibuprofen treatment induce morphological changes of the parotid tissues, which are related to macro- and micro-structure of the gland. PMID- 12653237 TI - Oral cancer: material deprivation, unemployment and risk factor behaviour--an initial study. AB - Regional variations in the incidence of oral cancer may be related in some cases to material deprivation. The main aim of this study was to identify in cases (and controls matched for age and sex) an index of material deprivation, employment history, smoking and alcohol habits. This prospective study comprised 100 subjects in each group and was questionnaire based. Previous studies in relation to material deprivation in oral cancer have been population based. There was a statistically significant trend for patients to come from the most deprived groups. Sixty-six per cent of the cases had experience of long-term unemployment. Although a high proportion, this was not statistically significant after multi variable analysis due to the confounding effects of smoking and alcohol use. Such a high proportion of cases with a history of long-term unemployment requires further study to examine in detail if there is a definite link with oral cancer once the confounding effects of smoking and alcohol are removed. PMID- 12653238 TI - Oral and maxillofacial tumours in north Jordanian children and adolescents: a retrospective analysis over 10 years. AB - This study evaluates the types and distribution of oral and maxillofacial tumours in north Jordanian children and adolescents. The records of the Department of Pathology at Jordan University of Science & Technology, during the period 1991 2000, were reviewed for patients younger than 19 years with oral and maxillofacial tumours. The tumours were analysed for age, sex, site and type. Out of all maxillofacial tumours, 23% occurred in children and adolescents, of these 10% were malignant and 90% were benign. 73% of the latter were soft tissue tumours and 17% were jaw tumours (58% odontogenic and 42% non-odontogenic). The most common benign soft tissue and jaw tumours were haemangioma and odontoma, respectively. 58% of malignant tumours were sarcomas and 42% were carcinomas. The mean age was 11 years with a female to male ratio of 1:1.2. The intraoral and extraoral sites most commonly affected by benign soft tissue tumours were the lower lip and face, respectively. While the total number of benign jaw tumours was slightly larger in the maxilla than the mandible, odontogenic tumours were more in the maxilla, and nonodontogenic tumours were more in the mandible. Differences found in this study between Jordanian children and adolescents and those from other countries may be attributable to genetic and geographic differences. The majority of benign tumours in the young are probably developmental rather than true neoplasms. PMID- 12653239 TI - Fourier and fractal analysis of maxillary alveolar ridge repair using platelet rich plasma (PRP) and inorganic bovine bone. AB - This report concerns the regeneration of the maxillary alveolar process in a 17 year-old patient who had lost the upper central incisors together with alveolar bone as a result of a car accident. Three months later, GBR (guided bone regeneration) was started with the use of autogenic platelet rich plasma (PRP) and inorganic bovine bone. The regenerated bone was analysed after 10 months and compared with intact bone using Fourier analysis of radiograms. The radial and spatial distribution of Fourier transforms showed that the original trabecular pattern existing in the intact bone on both sides of the defect was replicated in an evident way in the regenerated bone. Fractal analysis of intact and regenerated bone showed a higher fractal dimension for intact bone in comparison with regenerated bone, confirming a lower complexity of the newly formed trabecular structures. Replication of the original trabecular pattern in regenerated bone allows us to conclude that genetic mechanisms are influencing the organization of the trabecular pattern of regenerated bone tissue, probably under the influence of the growth factors contained in autologous PRP. PMID- 12653240 TI - Sinus lift approach for the retrieval of root fragments from the maxillary sinus. AB - A new application of a well-known technique; lateral wall sinus approach, to remove a root fragment from the maxillary sinus with minimal damage to the epithelial lining and anterior face of maxilla is described. The technique presented in this article helps eliminate some of the complications associated with Caldwell Luc procedure while preserving the epithelial lining of the maxillary sinus and bony face of maxilla. PMID- 12653241 TI - Bilateral chylothorax following radical neck dissection. AB - Bilateral chylothorax as a complication of radical neck dissection is extremely rare, but it is potentially serious and sometimes fatal. We found only 14 cases reported in the English literature. Here, we report a case of bilateral chylothorax following right modified and left radical neck dissections that was successfully treated with conservative management. PMID- 12653242 TI - Simultaneous repair of an orbital floor fracture and removal of an ethmoid osteoma: case report and review of the literature. AB - A case is presented of an ethmoid osteoma, which was discovered during the investigation of an orbital floor fracture. The osteoma was removed at the same time as orbital floor repair and special attention paid to preserving the medical canthal structures. PMID- 12653243 TI - Metastatic hepatocellular carcinoma of the maxillary sinus: a rare autopsy case without lung metastasis and a review. AB - A rare case of metastatic hepatocellular carcinoma (HCC) of the maxillary sinus in a 67-year-old man is reported along with the findings at autopsy. A fine needle aspiration biopsy specimen revealed a characteristic tumour structure resembling primary HCC. At autopsy, metastatic lesions were recognized in the bilateral adrenals, spleen, sternum, vertebrae and lymph node at the lesser curvature of the stomach, but not in the lung, suggesting that the HCC had metastasized to the maxillary sinus via the plexus venous vertebralis and/or the azygos vein, or lymph duct. In our reviewed 29 cases of metastatic HCC in the oro maxillofacial region, most patients were men in the 50- to 70-year age range. At least 11 cases did not have lung metastasis, and in 18, metastasis to the oro maxillofacial region was the first sign of HCC. One should be aware of the possibility to encounter the oral lesion as first sign of metastatic HCC. PMID- 12653244 TI - Tumour of the juxtaoral organ. AB - The juxtaoral organ is a normal and constant structure of the oral cavity. It consists of benign epithelial nests. We describe an intraoral tumour of the juxtaoral organ in a child. The tumour was not diagnosed after clinical and radiological examinations because it is extremely rare. A histological examination revealed a tumour of the juxtaoral organ, presumed to be neuroid hamartoma. This is only the second time that a tumour of the juxtaoral organ has been described in a child. We also describe the location, the embryology, the histology and the function of this organ. This is important because this structure can be confused with carcinomas of the oral cavity when examining frozen sections. PMID- 12653246 TI - Re: critical review on dry socket. PMID- 12653245 TI - Human dirofilariasis of the buccal mucosa: a case report. AB - Human dirofilariasis is a helminthic zoonosis that is common in some parts of the world. A sporadic case of dirofilariasis affecting the buccal mucosa has been reported in a non-endemic area of southern China. Clinical findings, diagnosis, pathogenesis and management are discussed. This is the fourth case of human dirofilariasis involving the oral mucosa reported in world literature. PMID- 12653247 TI - Patterning parameters associated with the branching of the ureteric bud regulated by epithelial-mesenchymal interactions. AB - The mechanisms by which the branching of epithelial tissue occurs and is regulated to generate different organ structures are not well understood. In this work, image analyses of the organ rudiments demonstrate specific epithelial branching patterns for the early lung and kidney; the lung type typically generating several side branches, whereas kidney branching was mainly dichotomous. Parameters such as the number of epithelial tips, the angle of the first branch, the position index of the first branch (PIFB) in a module, and the percentage of epithelial module type (PMT) were analysed. The branching patterns in the cultured lung and kidney, and in homotypic tissue recombinants recapitulated their early in vivo branching patterns. The parameters were applied to heterotypic tissue recombinants between lung mesenchyme and ureteric bud, and tip number, PIFB and PMT values qualified the change in ureter morphogenesis and the reprogramming of the ureteric bud with lung mesenchyme. All the values for the heterotypic recombinant between ureteric bud and lung mesenchyme were significantly different from those for kidney samples but similar to those of the lung samples. Hence, lung mesenchyme can instruct the ureteric bud to undergo aspects of early lung-type epithelial morphogenesis. Different areas of the lung mesenchyme, except the tracheal region, were sufficient to promote ureteric bud growth and branching. In conclusion, our findings provide morphogenetic parameters for monitoring epithelial development in early embryonic lung and kidney and demonstrate the use of heterotypic tissue recombinants as a model for studying tissue-specific epithelial branching during organogenesis. PMID- 12653248 TI - Xenopus neurula left-right asymmetry is respeficied by microinjecting TGF-beta5 protein. AB - A variety of TGF-beta-related ligands regulate the left-right asymmetry of vertebrates but the involvement of TGF-betas in left-right specification has not been reported. We assessed whether TGF-beta signaling is involved in the left right specification of Xenopus post-gastrula embryos by microinjecting Xenopus TGF-beta5 protein into the left or right flank of neurula-tailbud embryos. Injection on the right side of neurulae caused left-right reversal of the internal organs in 93% of the embryos, while injection on the left side caused less than 5% left-right reversal. Expression of Xenopus nodal related-1 (Xnr-1 ), Xenopus antivin and Xenopus Pitx2, which are normally expressed on the left, was unaltered by the left-side injection. In contrast, right-side injection into neurulae induced the expression of these genes predominantly on the right side. Right-side injection into tailbud embryos caused bilateral expression of these handed genes. Time course analysis of asymmetric gene expression revealed that Xnr-1 could be induced by TGF-beta5 at late neurula stage, while antivin and Pitx2 could be induced by TGF-beta5 at the latertail bud stage. Injection of the antisense morpholino oligonucleotide against Xenopus TGF-beta5 into the left dorsal blastomere inhibited the normal left-handed expression of Xnr-1 and Pitx2, and caused the organ reversal in the injected embryos. These results suggest that normal left-right balance of endogenous TGF-beta5 signaling in the neurula embryo may be needed to determine the laterality of the asymmetric genes and to generate the correct left-right axis. PMID- 12653249 TI - Developmental expression of Smad1-7 suggests critical function of TGF-beta/BMP signaling in regulating epithelial-mesenchymal interaction during tooth morphogenesis. AB - Members of the transforming growth factor-beta family (e.g. TGF-beta, BMP and activin) are critical regulators of tooth morphogenesis. The basic TGF-beta signaling engine consists of a receptor complex that activates Smads and a Smad containing complex that controls transcription of the downstream target genes. Little is known about the expression of endogenous Smads during tooth morphogenesis. Using a cRNA probe or antibody which specifically recognizes the expression of each Smad molecule, we provide a comprehensive endogenous Smad expression analysis during tooth morphogenesis. BMP signaling is transmitted through Smad1 and 5 which are first expressed within the dental lamina and later expand into condensed dental mesenchyme at the bud stage. As tooth development advances into the cap and bell stage, BMP signaling Smads are strongly localized within the inner enamel epithelium (IEE) and cranial neural crest derived dental mesenchyme (DM), indicating their critical role in regulating epithelial mesenchyme interaction during tooth morphogenesis. Smad2 and 3 are responsible for transmitting TGF-beta/activin signaling and show unique expression patterns during tooth morphogenesis. They are localized within the nuclei of both IEE and DM, suggesting that TGF-beta-activated Smads are critical for regulating tooth development. Smad4, the common Smad, is expressed in both dental epithelium and mesenchyme throughout all stages of tooth morphogenesis. The expression of inhibitory Smads (Smad6 and 7) largely overlaps with receptor regulated Smads, indicating that negative feedback on BMP/TGF-beta signaling is critical throughout all stages of tooth morphogenesis. Our results suggest that both receptor-regulated and inhibitory Smads are important regulators of tooth morphogenesis. The selective activation of Smad, as indicated by nucleartranslocation, may suggest selective activation of different members of the TGF-beta superfamily during tooth development. PMID- 12653250 TI - Genetic disruption of the growth hormone receptor does not influence motoneuron survival in the developing mouse. AB - In the rodent central nervous system (CNS) during the five days prior to birth, both growth hormone (GH) and its receptor (GHR) undergo transient increases in expression to levels considerably higher than those found postnatally. This increase in expression coincides with the period of neuronal programmed cell death (PCD) in the developing CNS. To evaluate the involvement of growth hormone in the process of PCD, we have quantified the number of motoneurons in the spinal cord and brain stem of wild type and littermate GHR-deficient mice at the beginning and end of the neuronal PCD period. We found no change in motoneuron survival in either the brachial or lumbar lateral motor columns of the spinal cord or in the trochlear, trigeminal, facial or hypoglossal nuclei in the brain stem. We also found no significant differences in spinal cord volume, muscle fiber diameter, or body weight of GHR-deficient fetal mice when compared to their littermate controls. Therefore, despite considerable in vitro evidence for GH action on neurons and glia, genetic disruption of GHR signalling has no effect on prenatal motoneuron number in the mouse, under normal physiological conditions. This may be a result of compensation by the signalling of other neurotrophic cytokines. PMID- 12653251 TI - CpZPC, a newt ZPC molecule, localizes to the inner surface of the egg envelope. AB - Zona pellucida-3 is an essential molecule for the binding of sperm to the egg envelope and for induction of the acrosome reaction in mice. Its homologous molecules, ZPCs, have been widely identified in the eggs of many vertebrates, except for urodeles. In this study, to investigate the participation of ZPC in newt fertilization, we cloned the cDNA of newt ZPC from the ovary of Cynops pyrrhogaster by reverse transcription polymerase chain reaction (RT-PCR). The cDNA was constructed from 1,397 nucleotides and included one open reading frame corresponding to a sequence of 439 amino acids. The deduced amino acid sequence had identities at 52, 47 and 45% with Xenopus gp41, mouse ZP3 and medaka L-SF, respectively. It included four potential N-linked glycosylation sites and 12 highly conserved cysteine residues of mammalian ZP3/ZPC molecules. This result suggests that CynopsZPC (cpZPC) has molecular features similar to those of mammalian ZP3/ZPCs. Messenger RNA for cpZPC was detected in the ovary and faintly in the testis. Two bands corresponding to 84 kDa and 70 kDa in the egg envelopes were detected by immunoblotting with an antiserum raised against a 9 amino acid peptide in the C-terminus domain of cpZPC. The molecular size of 84 kDa fits with the size of a putative sperm-binding protein reported by Nakai et al. (1999), suggesting that cpZPC may contribute to sperm binding to the egg envelope in C. pyrrhogaster. The results of immunohistochemistry suggest that cpZPC was localized in the inner surface of the egg envelope. Similar localization is seen only in fish, suggesting that cpZPC is a unique molecule which may allow us to investigate the functional evolution of the egg envelope in vertebrates. PMID- 12653252 TI - Disruption of eyelid and cornea development by targeted overexpression of the glucocorticoid receptor. AB - Glucocorticoid hormones act through the glucocorticoid receptor (GR) and they affect almost all physiological systems in the organism. We have previously reported that transgenic mice overexpressing GR under the control of the keratin k5 promoter (K5-GR mice) display severe phenotypic alterations in the epidermis and other ectoderm derivatives (Perez et al., 2001). In this work, we aimed to characterize the pathological consequences of GR targeted overexpression in the eyelid and cornea at late developmental stages. Despite glucocorticoids being widely prescribed as a topical treatment in ophthalmology, their potential role during ocular development in the embryo is not well understood. As shown by scanning electron microscopy analysis as well as by our histopathological and immunohistochemical data, long-term and newborn transgenic embryos showed unfused eyelids, along with proptosis of the globe and exposure of the anterior surface. In addition, epithelial defects were evident at the cornea. Our results indicate that GR overexpression affected the proliferation rate of targeted epithelia of the cornea and eyelid, thus demonstrating that GR was responsible for the arrest of epithelial proliferation of the developing eyelid edges, as well as for their destruction. We conclude that constitutive targeted overexpression of GR in the eyelid and corneal epithelium dramatically impairs ocular function in these transgenic mice. PMID- 12653253 TI - Distinct mechanisms underlie sperm-induced and protease-induced oolemma block to sperm penetration. AB - Fertilization of a mouse egg results in modification of the cytoplasmatic membrane (oolemma) which makes fusion with additional sperm impossible. CD9 is a transmembrane protein reported to be responsible for gamete fusion. Since the molecular mechanism of zygote membrane modification after fertilization remains unknown, we were interested to check whether lack of CD9 is the reason for non penetrability of zona-free zygotes. We wanted also to determine the effect of different methods of zona pellucida removal on the presence of CD9 on the surface of unfertilized eggs and their ability to be fertilized afterwards. We demonstrated that CD9 is present on the surface of both zygotes and parthenogenotes. We showed also that the treatment of eggs with pronase completely removes CD9 from the membrane of eggs making them infertile. Eggs treated with chymotrypsin and acid Tyrode still posses CD9 on their surface and remain fertile. The results of our experiments indicate that modification of the zygote oolemma does not involve a lack of CD9. We cannot exclude however, that the amount of CD9 decreases after fertilization. In addition, our studies indicate that the previously reported infertility of eggs treated with different proteases may result from the decrease or removal of CD9 and probably other proteins responsible for gamete fusion from the surface of eggs. PMID- 12653255 TI - Unequal distribution of otx1 mRnas among cleavage stage blastomeres in the teleost, Leucopsarion petersii (shiro-uo). AB - The otx genes belong to the orthodenticle gene family and play important roles in anterior brain development in vertebrates. We isolated two cDNA sequences, one homologous to human and zebrafish otxl and another homologous to zebrafish otx3, from the teleost Leucopsarion petersii (shiro-uo), which belongs to the family of gobies in the Perciformes. During embryogenesis of shiro-uo, otx1 and otx3 were expressed in the fore- and mid-brain throughout development in a manner similar to that observed in other vertebrates so far studied. However, otx-1 mRNA was also present at earlier stages and we obtained unique results using in situ hybridization and RT-PCR analysis demonstrating that otx-1 signals showed a distinct increase in the upper blastomeres, but not in the lower blastomeres, at the 8-cell stage. These stronger signals were maintained in the animal pole blastomeres during the 16-cell to 64-cell stages, followed by a gradual decrease during blastula stages. Such unexpected unequal distribution of otx1 mRNA revealed that blastomeres at early cleavage stages already showed non-equivalence in the embryogenesis of shiro-uo. PMID- 12653256 TI - Twenty-year follow-up of acquired renal cystic disease. AB - AIMS: Since 1979 the diseased kidneys of 96 patients on replacement therapy with chronic renal failure due to chronic glomerulonephritis have been followed to investigate the development of acquired cysts and tumors. This is a report of the 20-year follow-up. MATERIALS AND METHODS: Ninety-six patients were followed using periodic CT scan and were divided into hemodialysis, renal transplantation, bilateral nephrectomy and deceased groups during the follow-up. In the hemodialysis group, 36 patients (19 males, 17 females) were followed for 20 years. RESULTS: Kidney volumes which were 57.8 (1.51) (geometric mean (geometric SD)) ml at start of the follow-up had become 185.3 (2.03) ml 20 years later in males, and in females, 57.3 (1.64) ml had become 99.7 (2.36) ml. The increased rate was 3.2 (2.06) fold in males and 1.7 (2.57) fold in females. This enlargement of the kidneys was due to acquired cysts. Kidney volumes at the 20 year follow-up had increased more significantly than those at the 15-year follow up in males; however, kidney volumes at the 20-year follow-up had not changed in females, if compared with data at the 15-year follow-up. Kidney volumes in males at 20-year follow-up were significantly larger than those in females (p = 0.0232). Males with more than 3.2-fold in kidney volume increase at the 20-year follow-up were under the age of 40 at entry into this study (p = 0.0055), although the correlation between the degree of kidney volume increase and age was not significant (p = 0.0910). Kidney volumes in the transplantation group remained small. There was no new renal cell carcinoma development after 15-year follow-up except for the local recurrence of a previous operated case. Although 7 of 44 patients died during the past 20 years due to malignancy, no patient died of renal cell carcinoma because of early detection and treatment. One patient died of retroperitoneal bleeding, which is a complication of acquired renal cystic disease. CONCLUSION: Male preponderance of acquired cysts was maintained at the 20-year follow-up. There was a tendency for the rate of increase in acquired renal cystic disease to be larger in young males. No one died of renal cell carcinoma, although the incidence of renal cell carcinoma was high. PMID- 12653254 TI - 2P1, a novel male mouse cDNA specifically expressed during meiosis. AB - We screened a mouse germinal cell expression library with a probe derived from Sob1, a human testis-specific cDNA, and identified 2P1, a new mouse cDNA. A database search revealed that 2P1 was 91% identical to ORF1 of E3-3, a rat gene probably involved in the regulation of alternative splicing. Sequencing showed that 2P1 has a destabilization motif in its 3'-untranslated region. Northern blotting showed strong gene expression in the testis and weak expression in the epididymis, with no signal detected in other tissues. RT-PCR analysis confirmed testis and epididymis expression. In situ hybridization revealed that 2P1 mRNA was absent in spermatogonia but expressed in spermatocytes. This last result was confirmed by RT-PCR of FACS isolated primary spermatocytes (pachytene stage). Using RT-PCR, purified spermatids were also shown to express 2P1. PMID- 12653257 TI - Which factors account for renal stone formation in cystic fibrosis? AB - AIMS: Studies dealing with the increased tendency to stone formation noted in cystic fibrosis, focus on enteric hyperoxaluria. It is well recognized, however, that low urine volume, hypocitraturia and perhaps even hypercalciuria are further risk factors for stone formation. METHODS: Nineteen patients with cystic fibrosis (14 boys and 5 girls, aged 10-23, median 15 years) underwent a standard protocol for metabolic evaluation of the lithogenic tendency. In 10 patients, the study was repeated after treatment with recombinant human growth hormone 43 microgram/kg body weight daily for 12 months. RESULTS: The metabolic evaluation disclosed low urine output in 12, hyperoxaluria in 8 and hypocitraturia in 9 of the 19 cystic fibrosis patients. The mentioned parameters were not influenced by treatment with recombinant human growth hormone. CONCLUSION: The report indicates that in cystic fibrosis low urine volume, hypocitraturia and hyperoxaluria act in concert and contribute to the likelihood of stone formation. This tendency is not modified by treatment with recombinant human growth hormone. PMID- 12653258 TI - Combined effects of vasopressin V2 receptor antagonist and loop diuretic in humans. AB - AIMS: Patients with edematous diseases are generally treated with diuretics, but frequently present with hyponatremia associated with impaired water excretion. This study investigated the combined effects of vasopressin V2 receptor antagonist OPC-31260, an aquaretic, and furosemide, a saluretic, on free water excretion and serum sodium concentration (PNa). MATERIAL AND METHODS: The study was planned as an open-label, 4-period, 4-treatment, crossover study in 12 healthy subjects using single doses of placebo, furosemide 20 mg (F), OPC-31260 30 mg (O), and furosemide 20 mg plus OPC-31260 30 mg (O+F). RESULTS: OPC-31260 co administered with furosemide induced an increase in PNa and serum osmolality (F: 281.6 +/- 0.7, O+F: 284.9 +/- 0.7 mOsm/kgH2O) as a result of an additive increase in maximal urine volume (F: 7.7 +/- 1.9, O: 6.8 4 0.5, O+F: 13.2 +/- 1.4 ml/min) with increased electrolyte-free water excretion (F: 0.35 +/- 0.15, O: 5.3 4 0.5, O+F: 4.8 +/- 0.5 ml/min) while maintaining sodium excretion (F: 731 +/- 110, O+F: 1,064 +/- 149 microEq/min). It was suggested that by the co-administration, the resulting decreased generation of free water in Henle's loop was canceled out by a decrease in back--diffusion of free water across the collecting ducts. CONCLUSIONS: The co-administration may be particularly effective in treating edema and hyponatremia in edematous diseases. PMID- 12653259 TI - Renal ultrasonography is useful for evaluating diabetic renal failure. AB - AIM: It has been reported that the course of renal function is heterogeneous in patients with diabetic nephropathy. This study was undertaken to examine the clinical usefulness of renal ultrasonography in evaluating diabetic chronic renal failure (CRF) patients. METHODS: The renal sizes of type 2 diabetic patients with various degrees of renal injury, non-diabetic subjects without renal diseases and patients with non-diabetic CRF were measured by ultrasonography. The renal area index (RAI) was calculated from renal measurements and body surface area. The rate of renal function decline (delta l/cre) was analyzed by calculating the slope of the regression line for the reciprocal of serum creatinine concentrations over time. The correlations between delta l/cre and various clinical and laboratory parameters, including RAI, were analyzed. RESULTS: The RAI values of type 2 diabetic patients with nephropathy increased on the whole. It was also found that the RAI value of diabetic CRF patients was heterogeneous. There was a significant correlation between RAI and log delta l/cre (r = 0.492, p < 0.01). In addition to RAI, urinary protein excretion, serum albumin concentration and mean blood pressure significantly correlated with log delta l/cre. The correlation between RAI and log delta l/cre remained significant after adjustment for age, gender and serum albumin concentration. However, it was no longer significant after inclusion of mean blood pressure in the multivariate analysis. CONCLUSION: Although RAI is not a completely independent predictor of the risk of progression of diabetic renal failure, RAI could be a useful marker for the evaluation of diabetic renal failure. Renal involvement in diabetic patients is heterogeneous, and since renal ultrasonography is non-invasive and safe to perform, it is useful in evaluating diabetic CRF patients. PMID- 12653260 TI - Influence of hemodialysis on echocardiographic Doppler indices of the left ventricle: changes in parameters of systolic and diastolic function and Tei index. AB - AIMS: Numbers of previous studies have evaluated the influence of dialysis induced altered loading condition on Doppler-echocardiographic indices of left ventricle in patients with chronic renal failure. It has been suggested that most of Doppler-derived indices are preload-dependent. On the other hand, there are no studies that have evaluated the influence of hemodialysis on Tei index; a new Doppler-derived index obtained by isovolumetric contraction time plus isovolumetric relaxation time divided by ejection time. The aim of this study is to evaluate whether Tei index is also influenced by dialysis-induced altered loading condition as well as other Doppler-derived indices, and to assess the possibility that Tei index is also preload-dependent. PATIENTS AND METHODS: Thirty-two patients with chronic renal failure (21 men and 11 women, aged 48-93 years) on maintenance hemodialysis were evaluated for Doppler-derived indices before and after hemodialysis. We studied parameters of diastolic function (peak velocities of mitral inflow in early diastole (E) and late diastole from atrial filling (A), ratio of A to E (A/E), deceleration time (DT), and isovolumetric relaxation time (IRT)), parameters of systolic function (ejection time (ET), pre ejection period (PEP), ratio of PEP to ET (PEP/ET), and isovolumetric contraction time (ICT)) and Tei index. RESULTS: Hemodialysis resulted in significant decreases in E, increase in A/E, prolongation of IRT, no change in A and DT; significant prolongation of ICT and PEP, shortening of ET, and increase in PEP/ET and a significant increase in Tei index (0.42 +/- 0.16 vs 0.51 +/- 0.16, p < 0.0001). When patients were subdivided into 2 groups based on weight loss after hemodialysis (> or = 1.5 kg and < 1.5 kg), only the group that lost > or = 1.5 kg had significant change in Tei index before and after hemodialysis (0.40 +/- 0.15 vs 0.52 +/- 0.17, p = 0.0002). CONCLUSION: This study demonstrates that not only most of Doppler-derived indices but also Tei index is affected by dialysis induced altered loading condition and suggests that Tei index is possibly preload dependent. PMID- 12653261 TI - The effect of vitamin B12 on total plasma homocysteine concentration in folate replete hemodialysis patients. AB - AIM: Results from several studies indicate that the total homocysteine (tHcy) concentration in plasma is an independent risk factor for cardiovascular disease in hemodialysis patients. Folic acid is the established mainstay of homocysteine lowering treatment, but since such treatment does not normalize plasma tHcy concentration in hemodialysis patients, it is of importance to search for additional therapy. METHODS: Twenty-eight folate-replete hemodialysis patients were randomized to 2 equally sized groups, a treatment group and a control group. The treatment group received vitamin B12 tablets at a dose of 2 mg 3 times a week for 6 weeks (after each dialysis session) while the control group received no such treatment. Blood samples were collected before and at the end of the treatment period for analysis of tHcy in plasma and vitamin B12, methylmalonic acid as well as folate in serum. RESULTS: At the end of the study period, serum vitamin B12 concentrations were significantly higher in the treatment group than in the control group. Plasma tHcy concentrations decreased significantly in both groups during the study period. However, there was no difference between the responses of the 2 groups. CONCLUSION: The results of this open, randomized controlled study did not support the hypothesis that treatment with oral vitamin B12 has considerable homocysteine-lowering effect in folate-replete hemodialysis patients. PMID- 12653262 TI - Evaluation of bone mineral density after renal transplantation under a tacrolimus based immunosuppression: a pilot study. AB - BACKGROUND: Progressive bone loss consistently complicates renal transplantation (TP) in patients given an immunosuppression including prednisolone. The adjunction of cyclosporine in the immunosuppressive regimen does not reverse the negative impact of renal TP on the skeleton. The post-transplant effect of tacrolimus on bone mass is still unknown. METHODS: We evaluated the evolution of bone mineral density (BMD) and various biochemical markers over the first 12 months following renal TP in 23 patients given an immunosuppression combining tacrolimus and low-dose prednisolone. BMD of lumbar spine, total hip and hip subregions was measured by dual-energy X-ray absorptiometry within the first 15 days and 1 year after TP. RESULTS: At the time of TP, the average BMD was low in both the lumbar spine and the hip. After TP, a normalization of serum creatinine and a decrease in serum phosphate and iPTH levels occurs. Serum alkaline phosphatase level significantly rose transiently within the first 6 months and decreased thereafter. At 1 year post TP, BMD remained unchanged in the lumbar and in the trochanter subregions and rose in the other sites. BMD increased by at least 2% in 8, 13, 10 and 10 out of the 23 patients in the lumbar, neck, trochanter and total hip subregions, respectively. No correlation was found between evolution in BMD and age, sex, dialysis duration, level of hyperparathyroidism, prednisolone and tacrolimus cumulative intake and prescription of calcium, vitamin D or hormone replacement therapy. CONCLUSIONS: An immunosuppression combining tacrolimus and low-dose prednisolone might avoid the usual post-TP bone loss. Further randomized double-blind studies evaluating a larger cohort of patients should be undertaken to compare the effect of cyclosporine and tacrolimus on bone mass. PMID- 12653263 TI - Late onset seizures, hemiparesis and blindness in hemolytic uremic syndrome. AB - Neurologic complications of hemolytic uremic syndrome, including seizures, usually occur early during the acute phase of the illness. We report a3-year-old girl with classic diarrhea-associated hemolytic uremic syndrome who developed late onset seizures, hemiparesis and transient blindness on the 17th hospital day, at which time her recovery was characterized by improvement in her blood pressure, serum electrolytes, renal function, hematocrit and platelet count. A CT and MR revealed brainstem and posterior parietal and occipital infarct/edema. The association of these radiologic findings within the posterior distribution along with visual loss and seizures are unique to posterior reversible encephalopathy syndrome. Within 7 days, she regained motor function and vision and had no further seizure activity. At 6 months follow-up, physical examination revealed normal motor function and vision and a repeat MR showed near resolution of the previous findings with minimal occipital lobe gliosis. This case report describes the uncommon finding of late onset seizures occurring during the recovery phase of hemolytic uremic syndrome with MR findings consistent with posterior reversible encephalopathy syndrome. PMID- 12653264 TI - Steroid-induced tumor lysis syndrome in a patient with preleukemia. AB - Tumor lysis syndrome (TLS) is a well recognized complication of chemotherapy and radiotherapy for leukemia, lymphoma as well as rapidly growing malignancies. Less described is the occurrence of TLS following steroid therapy alone. Herein, we report on a 32-year-old male with myelodysplastic syndrome, characterized by refractory anemia with excess blasts in transformation, who developed acute oliguric renal failure 12 hours after methylprednisolone 1.0 g for presumed autoimmune thrombocytopenia. Laboratory investigations revealed typical findings of TLS, including hyperkalemia, marked hyperuricemia, hyperphosphotemia, hypocalcemia and urine uric acid to creatinine ratio 1.8 (> 1.0). Long hemodialysis (8 hours) was initiated for 3 consecutive sessions. Renal function recovered 1 week later. This case high-lights that single-dose steroid administration in a patient with hematological malignancy may cause the potential life-threatening complications of TLS. Prophylactic management prior to the use of steroid therapy for a variety of purposes is absolutely required in high-risk patients. PMID- 12653265 TI - Recurrent angiolymphoid hyperplasia with eosinophilia mimicking temporal arteritis associated with nephrotic syndrome. AB - We report on a middle-aged Caucasian male who presented with nephrotic syndrome that on 2 consecutive recurrences was accompanied by a pulsating tumor suggesting temporal arteritis. Renal biopsies showed features of a low-grade mesangial proliferative glomerulonephritis. The resected tumor in the temporal region revealed a lesion consistent with angiolymphoid hyperplasia with eosinophilia (ALHE), with moderate inflammatory involvement of the temporal artery. The patient was successfully treated with oral prednisolone in addition to removal of the tumor, but has remained steroid-dependent. To our knowledge, only 2 cases of ALHE and nephrotic syndrome have been reported so far in non-Japanese individuals [Altman et al. 1995, Sonkodi et al. 1987], and we are not aware of any previous case combining these features while simultaneuosly mimicking temporal arteritis. PMID- 12653266 TI - Efficacy of mycophenolate mofetil on recurrent glomerulonephritis after renal transplantation. AB - Recurrent glomerulonephritis in transplanted kidneys is not rare despite classical immunosuppressive drugs and depends on the etiology of nephropathy. Treatment of recurrence of renal disease on graft remains controversial. We report 6 cases of patients with recurrent glomerulonephritis after renal transplantation treated with mycophenolate mofetil (MMF). The glomerular diseases were Wegener's granulomatosis (n = 1), membranoproliferative glomerulonephritis type I (n = 1), focal and segmental glomerular sclerosis (n = 1), membranous glomerulonephritis (idiopathic membranous nephropathy (n = 1) and systemic lupus erythematous) (n = 1)) and immunoglobulin A nephropathy (n = 1). MMF was introduced because of intolerance of classical immunosuppressive treatment in 2 cases and because of its inefficiency in the other cases. MMF was introduced between 3 months and 36 months (13.5 +/- 7 months) after recurrence of the primitive glomerulonephritis. During combined MMF/cyclosporine/prednisone therapy, only 3 patients responded to MMF. MMF was disrupted precociously in 1 out of 3 patients who stabilized renal function because of discovery of lung cancer and in 2 out of the 3 other patients because of gastrointestinal intolerance and severe anemia. We supposed that MMF could represent a new effective alternative therapy of recurrent glomerulonephritis on renal graft in some cases. PMID- 12653267 TI - Hemodynamically relevant hematuria several months after biopsy of a kidney graft: an unusual cause. AB - We report the case of a 52-year-old female patient, who after a complicated living donor kidney transplantation, underwent kidney biopsy for suspected rejection. Duplex scanning revealed a small, asymptomatic arteriovenous (AV) fistula which was assessed as being hemodynamically unimportant. During follow up, several urinary tract infections occurred and recurrent short episodes of hematuria were attributed to cystitis, urethritis and urosepsis. Eight months later, the patient developed suddenly massive hematuria, tamponade of the urinary bladder and hemorrhagic shock as well as urosepsis. Duplex sonography showed a massive pseudoaneurysm in addition to the AV fistula. Arteriography confirmed the Duplex sonographic findings and embolization was performed after treatment of concomitant urosepsis. The fistula was closed completely and bleeding ceased. Although AV fistulas are rare complications of kidney biopsies and in most cases they remain asymptomatic, life-threatening hematuria can present several months after a biopsy due to the development of a pseudoaneurysm. Concomitant infectious complications of the urinary tract, bleeding disorders and other factors can be misleading during the assessment of the cause of gross hematuria. Regular Duplex sonographic follow-up examinations in patients with AV fistulas are advisable. PMID- 12653268 TI - Staghorn calculi complicating renal transplantation in patients with persistent post-transplantation hyperparathyroidism. AB - Renal stones rarely complicate renal transplants. Their causation is diverse. We describe 2 patients with significant staghorn calculi caused by metabolic factors. PMID- 12653269 TI - Successful retransplantation using rapamycin in a patient with previous calcineurin inhibitor-induced posterior leukoencephalopathy syndrome. AB - Posterior Leukoencephalopathy Syndrome (PLES) is a rare but serious neurological condition with many aetiologies. In the era of organ transplantation there have been sporadic reports of calcineurin-inhibitor associated PLES. We describe a case, with subsequent uneventful retransplantation using sirolimus. PMID- 12653270 TI - Successful perioperative blood purification therapy in patients with maintenance hemodialysis therapy who underwent living donor liver transplantation. AB - Living donor liver transplantation (LDLT) is a treatment for end-stage liver failure, and was developed to overcome the distinct insufficiency of cadaveric donors. Case 1 is a 56-year-old man who had undergone maintenance hemodialysis therapy for 4 years. An LDLT was performed for the treatment of advanced liver cirrhosis and hepatocellular carcinoma. Continuous hemodiafiltration (CHDF) was performed from the 2nd to 5th days after the operation. Case 2 is a 55-year-old man with primary amyloidosis and chronic renal failure. An LDLT was performed for the treatment of severe abdominal distention caused by a large liver volume. Although CHDF was started at the 3rd day after the operation, it was discontinued within 24 hours because of an increased urinary volume. CHDF was required again from the 6th-8th days, after which the blood purification mode was switched to regular intermittent hemodialysis. Meanwhile, no major problems occurred in either case. In conclusion, CHDF was required for about 5 days from the 2nd day after the operation. The application of careful and aggressive blood purification therapy during the perioperative period is a key to successful LDLT in dialysis patients. PMID- 12653271 TI - Behcet's disease and focal segmental glomerulosclerosis. PMID- 12653272 TI - A case of giant condylomata acuminata involving anus after renal transplantation. PMID- 12653273 TI - Acute hyponatremia and renal failure following percutaneous nephrolithotomy. PMID- 12653274 TI - Fractionation of Eucalyptus grandis chips by dilute acid-catalysed steam explosion. AB - Steam explosion of Eucalyptus grandis has been carried out under various pretreatment conditions (200-210 degrees C, 2-5 min) after impregnation of the wood chips with 0.087 and 0.175% (w/w) H2SO4. This study, arranged as a 2(3) factorial design, indicated that pretreatment temperature is the most critical variable affecting the yield of steam-treated fractions. Pretreatment of 0.175% (w/w) H2SO4-impregnated chips at 210 degrees C for 2 min was the best condition for hemicellulose recovery (mostly as xylose) in the water soluble fraction, reaching almost 70% of the corresponding xylose theoretical yield. By contrast, lower pretreatment temperatures of 200 degrees C were enough to yield steam treated substrates from which a 90% cellulose conversion was obtained in 48 h, using low enzyme loadings of a Celluclast 1.5 1 plus Novozym 188 mixture (Novo Nordisk). Release of water-soluble chromophores was monitored by UV spectroscopy and their concentration increased with pretreatment severity. The yield of alkali soluble lignin increased at higher levels of acid impregnation and pretreatment temperatures. Thermoanalysis of these lignin fractions indicated a pattern of lignin fragmentation towards greater pretreatment severities but lignin condensation prevailed at the most drastic pretreatment conditions. PMID- 12653275 TI - Rice straw-wood particle composite for sound absorbing wooden construction materials. AB - In this study, rice straw-wood particle composite boards were manufactured as insulation boards using the method used in the wood-based panel industry. The raw material, rice straw, was chosen because of its availability. The manufacturing parameters were: a specific gravity of 0.4, 0.6, and 0.8, and a rice straw content (10/90, 20/80, and 30/70 weight of rice straw/wood particle) of 10, 20, and 30 wt.%. A commercial urea-formaldehyde adhesive was used as the composite binder, to achieve 140-290 psi of bending modulus of rupture (MOR) with 0.4 specific gravity, 700-900 psi of bending MOR with 0.6 specific gravity, and 1400 2900 psi of bending MOR with a 0.8 specific gravity. All of the composite boards were superior to insulation board in strength. Width and length of the rice straw particle did not affect the bending MOR. The composite boards made from a random cutting of rice straw and wood particles were the best and recommended for manufacturing processes. Sound absorption coefficients of the 0.4 and 0.6 specific gravity boards were higher than the other wood-based materials. The recommended properties of the rice straw-wood particle composite boards are described, to absorb noises, preserve the temperature of indoor living spaces, and to be able to partially or completely substitute for wood particleboard and insulation board in wooden constructions. PMID- 12653276 TI - Semi-dry thermophilic anaerobic digestion of the organic fraction of municipal solid waste: focusing on the start-up phase. AB - The paper concerns the results of a pilot-scale study of the simulation of the start-up phase of the thermophilic semi-dry anaerobic digestion of the organic fraction of municipal solid wastes. The aim of the study was to aid and shorten the start-up phase of the full-scale plant (500 t/d) in Verona--Ca' del Bue, where the semi-dry anaerobic digestion process is being used. The substrate used in the experimentation was the mechanically sorted organic fraction of municipal solid waste (MS-OFMSW) enriched with the putrescent fraction from the source sorted OFMSW in order to simulate the substrate which is dealt with in the Verona plant. The results of the pilot scale study agreed with literature data and previous work of the authors: it showed a specific gas production of 0.23 m3/kg TVSfeed and a gas production rate of 2.1 m3/m3 d when operating at a specific organic loading rate of 0.135 kgTVSfeed/kgTVSreacter d. No problems regarding process stability were encountered in the gradual acclimation of the biomass. The design organic loading rate of 9 kg TVSfeed/m3reactor d was reached in about 30 days, during which the total solids content in the feedwas increased. Only a partial comparison with the full scale start-up, which is now in progress, is possible: this shows an initial general concordance with the results found in previous work. PMID- 12653277 TI - The influence of temperature and moisture contents regimes on the aerobic microbial activity of a biosolids composting blend. AB - To understand the relationships between temperature, moisture content, and microbial activity during the composting of biosolids (municipal wastewater treatment sludge), well-controlled incubation experiments were conducted using a 2-factor factorial design with six temperatures (22, 29, 36, 43, 50, and 57 degrees C) and five moisture contents (30, 40, 50, 60, and 70%). The microbial activity was measured as O2 uptake rate (mg g(-1) h(-1)) using a computer controlled respirometer. In this study, moisture content proved to be a dominant factor impacting aerobic microbial activity of the composting blend. Fifty percent moisture content appeared to be the minimal requirement for obtaining activities greater than 1.0 mg g(-1) h(-1). Temperature was also documented to be an important factor for biosolids composting. However, its effect was less influential than moisture content. Particularly, the enhancement of composting activities induced by temperature increment could be realized by increasing moisture content alone. PMID- 12653278 TI - Evaluation of Anammox and denitrification during anaerobic digestion of poultry manure. AB - Two approaches based on ne w process development and biological nitrogen transformation were investigated in a bench study for removing nitrogen as N2 gas from poultry waste while stabilizing the wastes. The process, known as "Anammox", was explored in batch anaerobic culture using serum bottles. The Anammox process involves the use of nitrite as an electron acceptor in the bacterially mediated oxidation of ammonia to yield N2. Studies are described wherein nitrite was added to poultry waste and the effects on ammonium levels were monitored. About 13-22% ammonium removal was observed with the inoculation of returned activated sludge, and the total ammonium reduction was not proportional to the reduction of nitrite, thereby suggesting that Anammox was less competitive under the conditions in our studies. The addition of nitrite and nitrate was not inhibitory to the process based on gas generation and COD reduction. The classical nitrogen removal process of nitrification followed with denitrification offers a more reliable basis for nitrogen removal from poultry wastes. PMID- 12653279 TI - Adsorption studies on rice husk: removal and recovery of Cd(II) from wastewater. AB - Adsorption behaviour of Ni(II), Zn(II), Cd(II) and Cr(VI) on untreated and phosphate-treated rice husk (PRH) showed that adsorption of Ni(II) and Cd(II) was greater when PRH was used as an adsorbent. Sorption of Cd(II) was dependent on contact time, concentration, temperature, adsorbent doses and pH of the solution. The Langmuir constants and thermodynamic parameters have been calculated at different temperatures. It was found that recovery of Cd(II) from synthetic wastewater by column operation was better than a batch process. PMID- 12653280 TI - Accumulation of copper by roots, hypocotyls, cotyledons and leaves of sunflower (Helianthus annuus L.). AB - The effects of different concentrations of copper sulfate on the growth of and the accumulation of Cu2+ by root, hypocotyl, cotyledon and leaf growth of sunflower (Helianthus annuus L.) were examined in this study. The concentrations of copper sulfate (CuSO4 x 5H2O) used were in the range from 10(-5) to 10(-3) M. Seedlings exposed to 10(-5) M Cu2+ solution exhibited a 33% increase in growth (P < 0.005) when compared with the root length of the control. The seedlings treated with 10(-3) M Cu2+ were significantly inhibited in shoot growth (P < 0.005). The Cu2+ content in roots, hypocotyls, cotyledons and leaves increased with increasing solution Cu2+ concentration. The roots of plants exposed to 10(-3) M Cu2+ accumulated a large amount of Cu (1070 microgram/g DW), and the Cu2+ level was approximately 25 fold higher than that of control. The Cu2+ contents in sunflower roots treated with 10(-4) and 10(-5) M Cu2+ were about 3.3 and 2.6 fold higher than the control, respectively. Also, the Cu2- level of the roots exposed to 10(-3) M Cu2+ was approximately 7.7 and 9.8 fold respectively, in comparison with the roots of plants grown in 10(-4) and 10(-5) M Cu2+. At 10(-3) M Cu2+, the Cu accumulated mainly in the roots (about 73%), and small amounts of Cu2+ (27%) were translocated to the hypocotyls, cotyledons and leaves. The Cu2+ concentration in the roots was less than that of the above parts of seedlings in treated groups with 10(-5) - 10(-4) M Cu2+. H. annuus has potential ability to accumulate Cu without being overly sensitive to Cu toxicity. PMID- 12653281 TI - Analysis of the logistic function model: derivation and applications specific to batch cultured microorganisms. AB - Mathematical models are useful for describing microbial growth, both in natural ecosystems and under research conditions. To this end, a rate expression that accounted for depletion of nutrients was used to derive the logistic function model for batch cultures. Statistical analysis was used to demonstrate the suitability of this model for growth curve data. Two linear forms of the model and two procedures for calculating growth rate constants were derived to facilitate statistical evaluation of growth curves. The procedures for calculating growth rate constants were found to be useful for calculation of growth rate constants at each time point, or for estimating growth rate constants from early growth curve data. The utility of the logistic function model and its alternative forms is discussed with respect to planning experiments, analyzing growth curves for the effects of factors other than nutrient limitation, and developing more complete descriptions of cell proliferation. PMID- 12653282 TI - Yield of Cassia angustifolia in combination with different tree species in a silvi-herbal trial under hot arid conditions in India. AB - A silvi-herbal trial was conducted in the hot arid region of India to study the performance of a shrub (Cassia angustifolia) in combination with different tree species. The study area was frequented by frost. Leaf yield of C. angustifolia under different treatments was estimated. It was found that the shrubs produced a significantly higher yield of leaves in the vicinity of the tree species as compared to the shrubs at a far distance from the trees. This was perhaps because of more protection of crops near the canopy of the plants during frosts. The yield was not much affected by the variation of tree species indicating that the effect of tree-shrub combination was not profound. C. angustifolia as inter-crop provides support to the farming system by way of conferring stability and generating assured income. PMID- 12653283 TI - Anaerobic degradation of No. 2 diesel fuel in the wetland sediments of Barataria Terrebonne estuary under various electron acceptor conditions. AB - The biodegradation of No. 2 diesel fuel under anaerobic conditions was investigated using sediments collected from wetlands of Barataria-Terrebonne estuary in Louisiana. The results indicated enhanced biodegradation of diesel fuel under sulfate-reducing, nitrate-reducing, methanogenic, and mixed electron acceptor conditions. However, the rate of diesel degradation was the highest under mixed electron acceptor conditions followed in order by sulfate-reducing, methanogenic, and nitrate-reducing conditions. Under mixed electron acceptor condition, 99% removal of diesel fuel was achieved within 510 days, while under sulfate-reducing condition 62% degradation of diesel fuel was observed for the same period. Diesel fuel was also degraded to a smaller extent in the culture condition where electron acceptors were not supplemented (natural attenuation condition). This study showed evidence for enhanced diesel fuel metabolism in a mixed microbial population system similar to any contaminated field site, where a heterogeneous microbial population exists. PMID- 12653284 TI - Survival of Salmonella spp. in a simulated acid-phase anaerobic digester treating sewage sludge. AB - The presence of pathogenic microorganisms in municipal waste sludge may create a serious outbreak of water borne diseases if the sludge is used for agricultural purpose. An attempt to decrease the number of pathogenic microorganisms, Salmonella spp. using a simulated acid-phase anaerobic digester was tested in a laboratory-scale batch experiment. Reduction of Salmonella spp. was demonstrated in a mixture of sludge and organic acid, simulating an acid digester of a two phase anaerobic digestion process. A high concentration of organic acid at a pH value of 5.5-6.0 prevents a decrease in Salmonella spp. concentration. Almost complete destruction of Salmonella spp. is observed within two days if the pH value is maintained below 5.5. PMID- 12653285 TI - Purification and characterization of extracellular phytase from Aspergillus niger ATCC 9142. AB - Extracellular phytase produced by Aspergillus niger ATCC 9142 was purified to homogeneity by employing an initial ultrafiltration step, followed by chromatography using ion exchange, gel filtration and chromatofocusing steps. The purified enzyme was an 84 kDa, monomeric protein. It possessed a temperature optimum of 65 degrees C, and a pH optimum of 5.0. Km and Vmax values of 100 microM and 7 nmol/s, respectively, were recorded and these values fall well within the range of those previously reported for microbial phytases. Substrate specificity studies indicated that, while the enzyme could hydrolyse a range of non-phytate-based phosphorylated substrates, its preferred substrate was phytate. Phytase activity was moderately stimulated in the presence of Mg2+, Mn2+, Cu2+, Cd2+, Hg2+, Zn2+ and F- ions. Activity was not significantly affected by Fe2- or Fe3- and was moderately inhibited by Ca2+. The enzyme displayed higher thermostability at 80 degrees C than did two commercial phytase products. Initial characterisation of the purified enzyme suggested that it could be a potential candidate for use as an animal feed supplement. PMID- 12653286 TI - Effects of Alternaria triticina and foliar fly ash deposition on growth, yield, photosynthetic pigments, protein and lysine contents of three cultivars of wheat. AB - A greenhouse experiment was conducted to study the effects of Alternaria triticina with and without foliar dusting of fly ash (0.0, 2.5, 5.0, 7.5 g plant( 1)/day(-1)) on the growth, yield, photosynthetic pigments, protein and lysine contents of three cultivars of wheat, Triticum aestivum. Dusting of 2.5 and 5.0 g fly ash caused a significant increase in growth, yield, photosynthetic pigments, protein and lysine contents of all the three cultivars. Dusting of 5.0 g fly ash caused a higher increase in the parameters than the 2.5 g dusting. However, dusting of 7.5 g fly ash had an adverse effect on growth, yield, photosynthetic pigments, protein and lysine contents. Cultivar HD-2009 suffered highest reductions in growth and yield and showed greater infected leaf area and disease symptoms from A. triticina followed by HD-2329 and Lok-1. Inoculation of A. triticina to plants dusted with 2.5/5.0 g fly ash gave higher reduction in growth and yield than did plants inoculated with A. triticina without fly ash. Cultivar Lok-1 showed highest growth, yield, photosynthetic pigments, protein and lysine contents followed by HD-2329 and HD-2009. PMID- 12653287 TI - A steady-state model for the evaluation of disk rotational speed influence on RBC kinetic: model presentation. AB - The physical and biological mechanisms of attached-biomass growth were analyzed and a steady-state model was proposed to determine the soluble carbonaceous removal in an RBC unit for different organic loading rates in the reactor. The objective of the model was the prediction of the organic loading rate corresponding to the maximum removal capacity in the system. A system of equations was solved where the influent soluble carbonaceous substrate concentration was the main variable. Monod's rate law was used for the growth of microorganism: the soluble carbonaceous substrate was the limiting substrate. Endogenous decay was neglected. The influence of disk rotational speed on the RBC removal capacity was investigated, the disk rotational speed being a parameter acting on oxygen transfer in the biofilm. The criteria for the evaluation of the kinetic parameter in the model were proposed. PMID- 12653288 TI - Manufacture of flame retardant foaming board from waste papers reinforced with phenol-formaldehyde resin. AB - A new flame retardant and waterproof porous fiberboard was manufactured from waste newspapers by using a foaming agent and a reinforcing phenol-formaldehyde resin. To find the optimum conditions for forming fiberboard with high mechanical properties as well as flame retardant and waterproof characteristics, a series of experiments were performed with varying weight percentages of resin (w). The results indicate that the best quality of the fiberboard was obtained for w approximately 11%. PMID- 12653289 TI - Characterization and viscosity parameters of seed oils from wild plants. AB - The physico-chemical properties of Spondias mombin seed oil and the viscosity temperature profiles of six seed oils from other plants which grow in the wild: Balanites aegytiaca, Lophira lanceolata, Sterculia setigera, Khaya senegalensis, Ximenia americana and Sclereocarya birrea, were investigated. The oil content of S. mombin seed was significant at 31.5% (w/w). The oil appeared stable as deduced from its low peroxide and acid values of 6.0 mEq kg(-1) and 1.68 mg KOH, respectively. The X. americana oil was denser than the other ones, with a value of 0.9625 g cm(-3) at 30 degrees C. The kinematic viscosities of the oils and their temperature dependence in the range 30-70 degrees C suggested a potential industrial application of the oils as lubricating base stock. Specifically, the kinematic viscosities of the oils were in the range 59.8-938.2 cst at 30 degrees C with X. americana having the highest value. At 70 degrees C, the reduction in viscosities of the oils was marked: reduction by over 70% of their values at 30 degrees C for S. setigera, K. senegalensis, X. americana and S. birrea oils. PMID- 12653290 TI - Impact of ozone mini-holes on the heterogeneous destruction of stratospheric ozone. AB - A comprehensive study of ozone mini-holes over the mid-latitudes of both hemispheres is presented, based on model simulations with the coupled climate chemistry model ECHAM4.L39(DLR)/CHEM representing atmospheric conditions in 1960, 1980, 1990 and 2015. Ozone mini-holes are synoptic-scale regions of strongly reduced total ozone, directly associated with tropospheric weather systems. Mini holes are supposed to have chemical and dynamical impacts on ozone levels. Since ozone levels over northern mid-latitudes show a negative trend of approximately 4%/decade and since it exists a negative correlation between total column ozone and erythemally active solar UV-radiation reaching the surface it is important to understand and assess the processes leading to the observed ozone decline. The simulated mini-hole events are validated with a mini-hole climatology based on daily ozone measurements with the TOMS (total ozone mapping spectrometer) instrument on the satellite Nimbus-7 between 1979 and 1993. Furthermore, possible trends in the event frequency and intensity over the simulation period are assessed. In the northern hemisphere the number of mini-hole events in early winter decreases between 1960 and 1990 and increases towards 2015. In the southern hemisphere a positive trend in mini-hole event frequency is detected between 1960 and 2015 in spring associated with the increasing Antarctic Ozone Hole. Finally, the impact of mini-holes on the stratospheric heterogeneous ozone chemistry is investigated. For this purpose, a computer-based detection routine for mini-holes was developed for the use in ECHAM4.L39(DLR)/CHEM. This method prevents polar stratospheric cloud formation and therefore heterogeneous ozone depletion inside mini-holes. Heterogeneous processes inside mini-holes amount to one third of heterogeneous ozone destruction in general over northern mid- and high-latitudes during winter (January-April) in the simulation. PMID- 12653291 TI - Theoretical consideration of methane emission from sediments. AB - A stationary theory of gas emission from sedimentary (active) layers of wetlands is developed. The theory takes into account methane generation in a sedimentary layer (W1) and its depth dependence, (W1(z)), the solubility, determined by Henry's constant (K1), and the mobility of methane molecules set by the methane diffusion coefficient (D1). The exponential dependence of methane generation rate decay with depth is considered in more detail. The penetration of atmospheric nitrogen into the active layer is also taken into consideration. It is shown that the value of diffusion methane flux from sedimentary layers is proportional to square root(K1D1P0W10) where P0 characterizes the atmospheric pressure and W10 characterizes the maximum generation rate. Coefficients relating the diffusion methane flux to the square root(K1D1P0W10) value are calculated for the different depth dependencies of methane generation rate. The values of these coefficients are not much different from unity for most real cases. PMID- 12653292 TI - Models to predict emissions of health-damaging pollutants and global warming contributions of residential fuel/stove combinations in China. AB - Residential energy use in developing countries has traditionally been associated with combustion devices of poor energy efficiency, which have been shown to produce substantial health-damaging pollution, contributing significantly to the global burden of disease, and greenhouse gas (GHG) emissions. Precision of these estimates in China has been hampered by limited data on stove use and fuel consumption in residences. In addition limited information is available on variability of emissions of pollutants from different stove/fuel combinations in typical use, as measurement of emission factors requires measurement of multiple chemical species in complex burn cycle tests. Such measurements are too costly and time consuming for application in conjunction with national surveys. Emissions of most of the major health-damaging pollutants (HDP) and many of the gases that contribute to GHG emissions from cooking stoves are the result of the significant portion of fuel carbon that is diverted to products of incomplete combustion (PIC) as a result of poor combustion efficiencies. The approximately linear increase in emissions of PIC with decreasing combustion efficiencies allows development of linear models to predict emissions of GHG and HDP intrinsically linked to CO2 and PIC production, and ultimately allows the prediction of global warming contributions from residential stove emissions. A comprehensive emissions database of three burn cycles of 23 typical fuel/stove combinations tested in a simulated village house in China has been used to develop models to predict emissions of HDP and global warming commitment (GWC) from cooking stoves in China, that rely on simple survey information on stove and fuel use that may be incorporated into national surveys. Stepwise regression models predicted 66% of the variance in global warming commitment (CO2, CO, CH4, NOx, TNMHC) per 1 MJ delivered energy due to emissions from these stoves if survey information on fuel type was available. Subsequently if stove type is known, stepwise regression models predicted 73% of the variance. Integrated assessment of policies to change stove or fuel type requires that implications for environmental impacts, energy efficiency, global warming and human exposures to HDP emissions can be evaluated. Frequently, this involves measurement of TSP or CO as the major HDPs. Incorporation of this information into models to predict GWC predicted 79% and 78% of the variance respectively. Clearly, however, the complexity of making multiple measurements in conjunction with a national survey would be both expensive and time consuming. Thus, models to predict HDP using simple survey information, and with measurement of either CO/CO2 or TSP/CO2 to predict emission factors for the other HDP have been derived. Stepwise regression models predicted 65% of the variance in emissions of total suspended particulate as grams of carbon (TSPC) per 1 MJ delivered if survey information on fuel and stove type was available and 74% if the CO/CO2 ratio was measured. Similarly stepwise regression models predicted 76% of the variance in COC emissions per MJ delivered with survey information on stove and fuel type and 85% if the TSPC/CO2 ratio was measured. Ultimately, with international agreements on emissions trading frameworks, similar models based on extensive databases of the fate of fuel carbon during combustion from representative household stoves would provide a mechanism for computing greenhouse credits in the residential sector as part of clean development mechanism frameworks and monitoring compliance to control regimes. PMID- 12653293 TI - The effect of long-term atmospheric CO2 enrichment on the intrinsic water-use efficiency of sour orange trees. AB - Every two months of 1992, as well as on three occasions in 1994-1995, we obtained leaf samples together with samples of surrounding air from eight well-watered and fertilized sour orange (Citrus aurantium L.) trees that were growing out-of-doors at Phoenix, Arizona, USA. These trees had been planted in the ground as small seedlings in July of 1987 and enclosed in pairs by four clear-plastic-wall open top chambers of which two have been continuously maintained since November of that year at a CO2 concentration of 400 micromol mol(-1) and two have been maintained at 700 micromol mol(-1). In September 2000, we also extracted north south and east-west oriented wood cores that passed through the center of each tree's trunk at a height of 45 cm above the ground. Stable-carbon isotope ratios (13C/12C) derived from these leaf, wood and air samples were used to evaluate each tree's intrinsic water-use efficiency (iWUE). The grand-average result was an 80% increase in this important plant parameter in response to the 300 micromol mol(-1) increase in atmospheric CO2 concentration employed in the study. This increase in sour orange tree iWUE is identical to the long-term CO2-induced increase in the trees' production of wood and fruit biomass, which suggests there could be little to no change in total water-use per unit land area for this species as the air's CO2 content continues to rise. It is also identical to the increase in the mean iWUE reported for 23 groups of naturally occurring trees scattered across western North America that was caused by the historical rise in the air's CO2 content that occurred between 1800 and 1985. PMID- 12653294 TI - Global eddy permitting simulations of surface ocean nitrogen, iron, sulfur cycling. AB - The geocycles of N, Fe and S in the ocean are tightly coupled and together exert strong influence on biogeochemistry of the earth system. We investigate this interaction by inserting macro-micronutrient cycling into a high resolution ocean model that realistically represents the general circulation. Simulated chlorophyll distributions reproduce many features of satellite-based measurements of ocean color. Meridional sections through model results and seasonally averaged SeaWiFS data agree reasonably well, both in the oligotrophic gyres and along fronts. Discrepancies are associated in many cases with shelf, ridge or island effects. Dimethyl sulfide peaks and their chlorophyll correlations are similar to those obtained on major oceanographic expeditions. Lack of strong regional relationships between the sulfide and phytoplankton may be partly explained by correspondence between time constants for production and for the traversal of mesoscale transport features. In general the eddies and filaments of tropical instability waves are well represented, including the onset of the 1997-1998 El Nino. North-south shifts of the transition zone chlorophyll front appear prominently in the results. PMID- 12653295 TI - Methane emission from fields with differences in nitrogen fertilizers and rice varieties in Taiwan paddy soils. AB - Flooded rice fields are one of the major biogenic methane sources. In this study, methane emission rates were measured after transplanting in paddy fields with application of two kinds of nitrogen fertilizers (ammonium sulfate, NH4+-N and potassium nitrate, NO3(-)-N) and with two kinds of rice varieties (Japonica and Indica). The experiment was conducted in fields located at Tainan District Agricultural Improvement Station in Chia-Yi county (23 degrees 25'08"N, 120 degrees 16'26"E) of southern Taiwan throughout the first and the second crop seasons in 1999. The seasonal methane flux in the first crop season with NH4+-N and NO3(-)-N ranged from 2.48 to 2.78 and from 8.65 to 9.22 g CH4 m(-2); and the values ranged 24.6-34.2 and 36.4-52.6 g CH4 m(-2) in the second crop season, respectively. In the first crop season, there were significantly increased 3.1 3.7-fold in methane emission fluxes due to plantation of Indica rice. In comparison of two rice varieties, the Indica rice variety showed a tendency for larger methane emission than the Japonica rice variety in the second crop season. Moreover, ammonium sulfate treatment significantly reduced CH4 emissions by 37 85% emissions compared to potassium nitrate plots. It was concluded that the CH4 emission was markedly dependent on the type of nitrogen fertilizer and rice variety in Taiwan paddy soils. PMID- 12653296 TI - Contribution of winter to the annual CH4 emission from a eutrophied boreal lake. AB - The springtime methane (CH4) emission from a small, eutrophied boreal lake was assessed during the winter ice-cover by measurement of gas ebullition and CH4 accumulation in the water column in association with the development of oxygen depletion after ice formation. The winter CH4 production was estimated to result in a loss of 3.6-7.9 g CH4 m(-2) from the lake to the atmosphere during the short period of ice melt. This could account for 22-48% of the annual CH4 emission from the pelagic zone of the lake. The contribution of winter to the annual CH4 release can be similar or even higher in seasonally ice-covered northern aquatic ecosystems than in northern terrestrial wetlands, thus winter must be considered in any studies into the aquatic CH4 emissions. The trophic state and wintertime oxygen conditions, linked to the changes in land-use in the catchments and climate, are important factors controlling the springtime lake CH4 emissions. PMID- 12653297 TI - Response to "bioterrorism" editorial. PMID- 12653298 TI - Infection rates of Amblyomma americanum and Dermacentor variabilis by Ehrlichia chaffeensis and Ehrlichia ewingii in southwest Missouri. AB - Both Ehrlichia chaffeensis and Ehrlichia ewingii are causative agents of human ehrlichiosis. Both pathogens are transmitted to humans through the bite of an infected lone star tick (Amblyomma americanum). Since Missouri has a high incidence of human monocytic ehrlichiosis, we investigated the prevalence of E. chaffeensis- and E. ewingii-infected A. americanum and Dermacentor variabilis (American dog tick) ticks to help assess the relative risk for humans exposed to these vectors. We used a nested polymerase chain reaction assay for the detection of ehrlichial DNA in the collected ticks. Infection rates for both ehrlichial species were calculated from the assay results for each of the tick species. E. chaffeensis was found to be present in 9.8% of adult A. americanum ticks (57 of 579) and 6.7% of D. variabilis ticks (eight of 120). E. ewingii DNA was present at an infection rate of 5.4% in adult A. americanum (31 of 579) and 3.3% of D. variabilis ticks (four of 120). A minimum infection rate for nymph pools of A. americanum was 1.7% for E. chaffeensis and 0.6% for E. ewingii. PMID- 12653299 TI - Experimental evaluation of rodent exclusion methods to reduce hantavirus transmission to residents in a Native American community in New Mexico. AB - We conducted a pilot study to evaluate the efficacy of rodent proofing continuously occupied homes as a method for lowering the risk for hantavirus pulmonary syndrome (HPS) among residents of a Native American community in northwestern New Mexico. Rodent proofing of dwellings was paired with culturally appropriate health education. Seventy homes were randomly assigned to treatment or control categories. Treatment homes were rodent-proofed by sealing openings around foundations, doors, roofs, and pipes and repairing screens and windows. Repairs to each dwelling were limited to $500 US. After repairs were completed, 15-20 snap traps were placed in each treatment and control home and checked approximately every 2 days for an average of 3-4 weeks. During 23,373 trap nights, one house mouse (Mus musculus) was captured in one treatment home, and 20 mice (16 deer mice, Peromyscus maniculatus, two Pinyon mice, Peromyscus truei, and two unidentified mice) were captured in five control homes (one house had 14 captures, two had two captures, and two had one capture). Trap success was 0.01% in treatment homes and 0.15% in controls. Intensity of infestation (mean number of mice captured per infested home) was 1 in treatment homes and 4 in controls. Observations of evidence of infestation (feces, nesting material, gnaw marks, or reports of infestation by occupant) per 100 days of observation were 1.2 in treatment homes and 3.1 in controls. Statistical power of the experiment was limited because it coincided with a period of low rodent abundance (August November 2000). Nevertheless, these results suggest that inexpensive rodent proofing of occupied rural homes can decrease the frequency and intensity of rodent intrusion, thereby reducing the risk of HPS among rural residents in the southwestern United States. PMID- 12653300 TI - Identification of Ctenocephalides felis fleas as a host of Rickettsia felis, the agent of a spotted fever rickettsiosis in Yucatan, Mexico. AB - In search for the vector of the recently recognized spotted fever rickettsiosis of the Yucatan, ticks, fleas, and lice were collected from vegetation and dogs in localities where seropositive persons had been found. The arthropods were examined by polymerase chain reaction (PCR) using primers for the genus-specific 17-kDa protein gene followed by restriction fragment length polymorphism (RFLP) and DNA sequencing. Eleven (20%) of 54 pools of Ctenocephalides felis fleas contained DNA of Rickettsia felis. None of 219 Amblyomma cajennense, 474 Rhiphicephalus sanguineus, 258 Boophilus sp. ticks, and 33 Poliplax species lice contained DNA of Rickettsia. The identity of the rickettsial DNA was confirmed as R. felis by PCR/RFLP for the citrate synthase and outer membrane protein A genes and by DNA sequencing. The results indicate that the host of R. felis in Yucatan is C. felis and suggest that the spotted fever rickettsiosis that has infected >5% of the population of the Yucatan and can present as a dengue-like illness is likely to be caused by R. felis. PMID- 12653301 TI - Spatiotemporal analysis of epizootic raccoon rabies propagation in Connecticut, 1991-1995. AB - The quantitative analysis of pathogen transmission within its specific spatial context should improve our ability to predict and control the epizootic spread of that disease. We compared two methods for calibrating the effect of local, spatially distributed environmental heterogeneities on disease spread. Using the time-of-first-appearance of raccoon rabies across the 169 townships in Connecticut, we estimated local spatial variation in township-to-township transmission rate using Trend Surface Analysis (TSA) and then compared these estimates with those based on an earlier probabilistic simulation using the same data. Both the probabilistic simulation and the TSA reveal significant reduction in transmission when local spatial domains are separated by rivers. The probabilistic simulation suggested that township-to-township transmission was reduced sevenfold for townships separated by a river. The global effect of this sevenfold reduction is to increase the time-to-first-appearance in the eastern townships of Connecticut by approximately 29.7% (spread was from west to east). TSA revealed a similar effect of rivers with an overall reduction in rate of local propagation due to rivers of approximately 22%. The 7.7% difference in these two estimates reveals slightly different aspects of the spatial dynamics of this epizootic. Together, these two methods can be used to construct an overall picture of the combined effects of local spatial variation in township-to township transmission on patterns of local rate of propagation at scales larger than the immediate nearest neighboring townships. PMID- 12653302 TI - Investigation of the southern limits of Murray Valley encephalitis activity in Western Australia during the 2000 wet season. AB - Western Australia experienced its worst-ever outbreak of the mosquito-borne Murray Valley encephalitis (MVE) virus during the 2000 wet season. Highest-on record rainfall throughout much of the state during the 2000 wet season gave rise to extensive mosquito breeding and increased MVE virus transmission, resulting in nine cases of encephalitis. Activity of MVE virus in Western Australia is monitored by detecting MVE virus-specific antibodies in serum from sentinel chickens, located at towns and communities throughout the north of the state. However, during 2000, all 28 flocks of chickens seroconverted to MVE virus, including a flock located >600 km further south than MVE virus activity had ever previously been recorded. Furthermore, the majority of the nine cases of encephalitis occurred outside the enzootic Kimberley region. We therefore undertook a major serosurvey of domestic chicken flocks both south and east of the previously defined regions of virus activity. The results suggest that MVE virus activity extended as far south as the Midwest and northern Goldfields during 2000. A new southern limit of activity of MVE virus is therefore proposed. The results have implications for managing outbreaks of MVE virus in Western Australia and have enabled us to locate additional sentinel flocks as part of the MVE surveillance program for future years. PMID- 12653303 TI - The impact of ecological conditions on the prevalence of malaria among orangutans. AB - Contemporary human land use patterns have led to changes in orangutan ecology, such as the loss of habitat. One management response to orangutan habitat loss is to relocate orangutans into regions of intact, protected habitat. Young orangutans are also kept as pets and have at times been a valuable commodity in the illegal pet trade. In response to this situation, government authorities have taken law enforcement action by removing these animals from private hands and attempted to rehabilitate and release these orangutans. In relocating free ranging orangutans, the animals are typically held isolated or with family members for <48 h and released, but during the course of rehabilitation, orangutans often spend some time in captive and semicaptive group settings. Captive/semicaptive groups have a higher density of orangutans than wild populations, and differ in other ways that may influence susceptibility to infectious disease. In order to determine the impact of these ecological settings on malaria, the prevalence of malaria was compared between 31 captive and semicaptive orangutans in a rehabilitation program at the Sepilok Orangutan Rehabilitation Centre and 43 wild orangutans being moved in a translocation project. The prevalence of malaria parasites, as determined by blood smear and Plasmodium genus-specific nested-polymerase chain reaction, was greater in the captive/semicaptive population (29 of 31) than in the wild population (5 of 43) even when accounting for age bias. This discrepancy is discussed in the context of population changes associated with the management of orangutans in captive/semicaptive setting, in particular a 50-fold increase in orangutan population density. The results provide an example of how an ecological change can influence pathogen prevalence. PMID- 12653304 TI - A recombinant envelope protein-based enzyme-linked immunosorbent assay for West Nile virus serodiagnosis. AB - Recombinant West Nile virus envelope (E) protein was examined in enzyme-linked immunosorbent assay (ELISA) to detect antibodies elicited during West Nile virus infection. Horses (nine of 10) and humans (six of six) with confirmed West Nile virus infection had IgG and/or IgM antibodies to the E protein. Antibodies to the recombinant West Nile virus membrane and nonstructural 1 proteins were not detected in any of these sera. An E protein-based ELISA may aid in the serological diagnosis of West Nile virus infection. PMID- 12653305 TI - Occurrence of Campylobacter spp. and Cryptosporidium spp. in seagulls (Larus spp.). AB - An investigation was carried out into the prevalence of thermophilic Campylobacter subspecies (spp.) and Cryptosporidium spp. in fresh fecal specimens collected from members of the gull family (Larus spp.) from three coastal locations of Northern Ireland. A total of 205 fresh fecal specimens were collected from gulls, of which 28 of 205 (13.7%) were positive for Campylobacter spp. and none of 205 for Cryptosporidium spp. Of these campylobacters, 21 of 28 (75%) isolates obtained belonged to the urease-positive thermophilic Campylobacter (UPTC) taxon, followed by five of 28 (17.9%) Campylobacter lari and 2/28 (7.1%) Campylobacter jejuni. It is significant that seagulls are the sole warm-blooded animal host of this bacterial taxon in Northern Ireland. It is proposed that physiological adaptation to starvation by gulls may lead to increased concentrations of urea through energy production from protein, yielding increased levels of urea for metabolism by UPTC organisms. In general, the possibility exists that environmental contamination of surface waters with campylobacters might be mediated by wild birds (such as gulls), where such waters are used for recreational purposes or where such waters are consumed untreated, might represent a risk to public health. PMID- 12653306 TI - Zoonoses--an interview with James Emory Childs, Sc.D. PMID- 12653307 TI - Chronology of primate discoveries in Myanmar: influences on the anthropoid origins debate. AB - The history of primate paleontology in Asia is long and complex, beginning with the first discoveries of fossil primates on the Indian subcontinent in the early 1830's. The first Eocene mammals from Asia were collected in Myanmar and described in 1916, while the first primates, Pondaungia and Amphipithecus, were described in 1927 and 1937, respectively, both from the Pondaung Formation in Myanmar. For the next 60 years, these two Pondaung taxa remained as the only known Eocene primates from Myanmar and one of the few records of Eocene primates from all of Asia. Taxonomically, Pondaungia and Amphipithecus were linked with a number of different groups, including archaic, hoofed ungulates (condylarths), adapiform primates, omomyid primates, and anthropoids. While no consensus existed, Pondaungia and Amphipithecus were most commonly compared with anthropoids. Beginning in the late 1990s, new primates were discovered in Myanmar, including smaller-bodied forms such as Bahinia and Myanmarpithecus. Also, new and better specimens of the larger-bodied Pondaungia and Amphipithecus began to appear, including the first cranial and postcranial fragments. Evaluations based on these new specimens, especially the postcrania, indicate that the two larger-bodied Myanmar taxa are adapiform primates that show their closest affinities to North American notharctines. The smaller-bodied taxa remain enigmatic, but may share their closest affinities with North American and Asian omomyid primates and Asian Tarsius. None of the known Asian primate taxa appear closely related to African anthropoids, which suggests that true anthropoids did not reach Asia until the latest Oligocene or earliest Miocene. These facts make an Asian origin for Anthropoidea unlikely. Additional and earlier evidence from both Asia and Africa is required before the ultimate origin of anthropoids can be determined. It appears possible that true anthropoids were an ancient radiation that may have been part of a Gondwanan (southern hemisphere) community that is, at present, poorly sampled and little understood. PMID- 12653308 TI - On the nature and evolution of the neural bases of human language. AB - The traditional theory equating the brain bases of language with Broca's and Wernicke's neocortical areas is wrong. Neural circuits linking activity in anatomically segregated populations of neurons in subcortical structures and the neocortex throughout the human brain regulate complex behaviors such as walking, talking, and comprehending the meaning of sentences. When we hear or read a word, neural structures involved in the perception or real-world associations of the word are activated as well as posterior cortical regions adjacent to Wernicke's area. Many areas of the neocortex and subcortical structures support the cortical striatal-cortical circuits that confer complex syntactic ability, speech production, and a large vocabulary. However, many of these structures also form part of the neural circuits regulating other aspects of behavior. For example, the basal ganglia, which regulate motor control, are also crucial elements in the circuits that confer human linguistic ability and abstract reasoning. The cerebellum, traditionally associated with motor control, is active in motor learning. The basal ganglia are also key elements in reward-based learning. Data from studies of Broca's aphasia, Parkinson's disease, hypoxia, focal brain damage, and a genetically transmitted brain anomaly (the putative "language gene," family KE), and from comparative studies of the brains and behavior of other species, demonstrate that the basal ganglia sequence the discrete elements that constitute a complete motor act, syntactic process, or thought process. Imaging studies of intact human subjects and electrophysiologic and tracer studies of the brains and behavior of other species confirm these findings. As Dobzansky put it, "Nothing in biology makes sense except in the light of evolution" (cited in Mayr, 1982). That applies with as much force to the human brain and the neural bases of language as it does to the human foot or jaw. The converse follows: the mark of evolution on the brains of human beings and other species provides insight into the evolution of the brain bases of human language. The neural substrate that regulated motor control in the common ancestor of apes and humans most likely was modified to enhance cognitive and linguistic ability. Speech communication played a central role in this process. However, the process that ultimately resulted in the human brain may have started when our earliest hominid ancestors began to walk. PMID- 12653309 TI - The promise of geometric morphometrics. AB - Nontraditional or geometric morphometric methods have found wide application in the biological sciences, especially in anthropology, a field with a strong history of measurement of biological form. Controversy has arisen over which method is the "best" for quantifying the morphological difference between forms and for making proper statistical statements about the detected differences. This paper explains that many of these arguments are superfluous to the real issues that need to be understood by those wishing to apply morphometric methods to biological data. Validity, the ability of a method to find the correct answer, is rarely discussed and often ignored. We explain why demonstration of validity is a necessary step in the evaluation of methods used in morphometrics. Focusing specifically on landmark data, we discuss the concepts of size and shape, and reiterate that since no unique definition of size exists, shape can only be recognized with reference to a chosen surrogate for size. We explain why only a limited class of information related to the morphology of an object can be known when landmark data are used. This observation has genuine consequences, as certain morphometric methods are based on models that require specific assumptions, some of which exceed what can be known from landmark data. We show that orientation of an object with reference to other objects in a sample can never be known, because this information is not included in landmark data. Consequently, a descriptor of form difference that contains information on orientation is flawed because that information does not arise from evidence within the data, but instead is a product of a chosen orientation scheme. To illustrate these points, we apply superimposition, deformation, and linear distance-based morphometric methods to the analysis of a simulated data set for which the true differences are known. This analysis demonstrates the relative efficacy of various methods to reveal the true difference between forms. Our discussion is intended to be fair, but it will be obvious to the reader that we favor a particular approach. Our bias comes from the realization that morphometric methods should operate with a definition of form and form difference consistent with the limited class of information that can be known from landmark data. Answers based on information that can be known from the data are of more use to biological inquiry than those based on unjustifiable assumptions. PMID- 12653310 TI - Ancient DNA in anthropology: methods, applications, and ethics. AB - Anthropologists were quick to recognize the potential of new techniques in molecular biology to provide additional lines of evidence on questions long investigated in anthropology, as well as those questions that, while always of interest, could not have been addressed by more traditional techniques. The earliest ancient DNA studies, both within anthropology and in other fields, lacked rigorous hypothesis testing. However, more recently the true value of ancient DNA studies is being realized, and methods are being applied to a wide variety of anthropological questions. We review the most common methods and applications to date, and describe promising avenues of future research. We find that ancient DNA analyses have a valuable place in the array of anthropological techniques, but argue that such studies must not be undertaken merely to demonstrate that surviving DNA is present in organic remains, and that no such work should be performed before a careful consideration of the possible ethical ramifications of the research is undertaken. PMID- 12653313 TI - Interpreting the posture and locomotion of Australopithecus afarensis: where do we stand? AB - Reconstructing the transition to bipedality is key to understanding early hominin evolution. Because it is the best-known early hominin species, Australopithecus afarensis forms a baseline for interpreting locomotion in all early hominins. While most researchers agree that A. afarensis individuals were habitual bipeds, they disagree over the importance of arboreality for them. There are two main reasons for the disagreement. First, there are divergent perspectives on how to interpret primitive characters. Primitive traits may be retained by stabilizing selection, pleiotropy, or other ontogenetic mechanisms. Alternately, they could be in the process of being reduced, or they simply could be selectively neutral. Second, researchers are asking fundamentally different questions about the fossils. Some are interested in reconstructing the history of selection that shaped A. afarensis, while others are interested in reconstructing A. afarensis behavior. By explicitly outlining whether we are interested in reconstructing selective history or behavior, we can develop testable hypotheses to govern our investigations of the fossils. To infer the selective history that shaped a taxon, we must first consider character polarity. Derived traits that enhance a particular function, are found to be associated with that function in extant homologs, and that epigenetically sensitive data indicate were actually being used for that function, can be interpreted as adaptations. The null hypothesis to explain the retention of primitive traits is that of selective neutrality, or nonaptation. Disproving this requires demonstration of active stabilizing or negative selection (disaptation). Stabilizing selection can be inferred when primitive traits compromise a derived function clearly of adaptive value. Prolonged stasis, continued use of the trait for a particular function, or no change in variability in the trait are evidence that can support a hypothesis of adaptation for primitive traits, but still do not falsify the null hypothesis. Disaptation, or negative selection, should result in a trait being reduced or lost. To infer the behaviors of a fossil species, we must first determine its adaptations, use this to make hypotheses about its behavior, and test these hypotheses using epigenetically sensitive traits that are modified by an individual's activity pattern. When the A. afarensis data are evaluated using this framework, it is clear that these hominins had undergone selection for habitual bipedality, but the null hypothesis of nonaptation to explain the retention of primitive, ape-like characters cannot be falsified at present. The apparent stasis in Australopithecus postcranial form is currently the strongest evidence for stabilizing selection maintaining its primitive features. Evidence from features affected by individual behaviors during ontogeny shows that A. afarensis individuals were habitually traveling bipedally, but evidence presented for arboreal behavior so far is not conclusive. By clearly identifying the questions we are asking about early hominin fossils, refining our knowledge about character polarities, and elucidating the factors influencing morphology, we will be able to progress in our understanding of the posture and locomotion of A. afarensis and all early hominins. PMID- 12653311 TI - Canalization, developmental stability, and morphological integration in primate limbs. AB - Canalization and developmental stability refer to the tendency of developmental processes to follow particular trajectories, despite external or internal perturbation. Canalization is the tendency for development of a specific genotype to follow the same trajectory under different conditions (different environments or different genetic backgrounds), while developmental stability is the tendency for the development of a specific genotype to follow the same trajectory under the same conditions. Morphological integration refers to the tendency for structures to show correlated variation because they develop in response to shared developmental processes or function in concert with other structures. All three phenomena are emergent properties of developmental systems that can affect the interaction of development and evolution. In this paper, we review the topics of canalization, developmental stability, and morphological integration and their relevance to primate and human evolution. We then test three developmentally motivated hypotheses about the patterning of variability components in the mammalian limb. We find that environmental variances and fluctuating asymmetries (FA) increase distally along the limb in adult macaques but not in fetal mice. We infer that the greater variability of more distal segments in macaques is due to postnatal mechanical effects. We also find that heritability and FA are significantly correlated when different limb measurements are compared in fetal mice. This supports the idea that the mechanisms underlying canalization and developmental stability are related. Finally, we report that the covariation structure of fore- and hindlimb skeletal elements shows evidence for morphological integration between serially homologous structures between the limbs. This is evidence for the existence of developmental modules that link structures between the limbs. Such modules would produce covariation that would need to be overcome by selection for divergence in hind- and forelimb morphology. PMID- 12653314 TI - Temporization for endodontics. AB - Teeth undergoing root-canal therapy are susceptible to microbial contamination from oral fluids both during and after treatment. With the exception of single visit treatment, the use of a temporary restoration is mandatory. This review aims to provide an overview of the materials and techniques used for short- and long-term restorations during and immediately after endodontic treatment, and to make clinical recommendations. Further research is necessary to determine the effectiveness of temporary restorations in the conditions of the oral environment, especially with respect to leakage and functional demands. PMID- 12653312 TI - Critical periods in human growth and their relationship to diseases of aging. AB - It has long been recognized that there are "critical periods" during mammalian development when exposure to specific environmental stimuli are required in order to elicit the normal development of particular anatomical structures or their normal functioning. The responses of the organism to these stimuli depend on a specific level of anatomical maturation and a state of rapid anatomical and/or functional change. This discussion of critical periods in growth is not confined to the classic definition of a narrow time frame of development during which a particular environmental threshold or limit must exist for normal growth and function to ensue. Using both auxological and epidemiological approaches, we suggest a lifespan perspective which encompasses accumulating and interacting risks that are manifest from prenatal life onward. By understanding the process of growth development, and by scrutinizing the growth process, early variations that lead to later disease can be identified. Here we review a significant amount of the evidence that links exposure during growth to later morbidity and mortality. The fetus appears to respond to insults during the prenatal period through the process of "programming," which has short-term survival advantages but may have a long-term disadvantage in that it is associated with cardiovascular disease, hypertension, type II diabetes, and later obesity. Low birth weight combined with rapid postnatal growth during infancy also appears to be associated, for instance, with later childhood and adult sequelae in terms of glucose tolerance and obesity. Independent of birth weight, the timing of adiposity rebound during mid-childhood also predicts later obesity. The timing, magnitude, and duration of adolescent growth and maturationare associated with critical body composition changes, including the normal acquisition of body fat and bone mineralization. In particular, the acquisition of appropriate peak bone mass is critical in determining the later risk of osteoporosis. A putative causal mechanism linking early growth variation to later chronic disease risk through telomeric attrition is discussed. The obligatory loss of telomeric DNA with each cell division serves as a mitotic clock and marks the rate of growth and repair processes in the cell. Although much more work is required, existing studies support the notion that telomere shortening is not only a clock of cellular division, but also marks relative growth rate, as well as contributing to common degenerative processes of aging through its impact on cellular senescence. PMID- 12653315 TI - A quantitative evaluation of apical leakage of four root-canal sealers. AB - AIM: This in vitro study evaluated the microleakage of root fillings involving four root-canal sealers including AH Plus (De-Trey, Switzerland), RoekoSeal (Roeko, Germany). Ketac-Endo (ESPE, (Germany) and Sultan (Sultan Chemists, USA. METHODOLOGY: Forty extracted human maxillary anterior teeth were used. The coronal part of each tooth was removed and the root canals were prepared using the step-back technique and apical enlargement to size 60 file. The specimens were randomly divided into four groups of 10 samples, filled with one of the test materials and gutta-percha cones by the cold lateral condensation technique. A fluid filtration method was used for quantitative evaluation of apical leakage. The leakage was measured by the movement of an air bubble in a capillary glass tube connected to the experimental root section. Measurements of fluid movement were made at 2-min intervals for 8 min. Water transport through existing voids in the obturated canals could be measured reproducibly in this way. The quality of the seal of each specimen was measured after 7,14 and 21 days. RESULTS: Statistical analysis indicated that all the root fillings showed less leakage after 21 days (P < 0.05). Fillings incorporating Sultan showed significantly more leakage than all other sealers. CONCLUSIONS: Root fillings with RoekoSeal in combination with cold lateral condensation technique showed better sealing than those with Ketac-Endo, AH Plus and Sultan sealers after 21 days. The fluid filtration test used in this study gave quantitative results and allowed nondestructive long-term evaluation of specimens. PMID- 12653317 TI - Indirect digital images: limit of image compression for diagnosis in endodontics. AB - AIM: To analyse the impact of the loss of information that results from the compression of a file containing a radiographic image. METHODOLOGY: Fourteen intraoral radiographs were digitized employing an AGFA ARCUS II scanner at an optical resolution of 300 ppi and with a grey scale of 8 bytes. The images were stored in Tagged Image File Format (TIFF). The files were compressed with no information loss with the WinZip 8.0 program. Compression with information loss was performed using the Photoshop 5.0 program (Adobe Systems Inc., San Jose, CA. USA) and the Joint Photograph Expert (Group (JPEG) format. Each of the images was compressed to one of the 11 qualities available (10-0). An expert performed the qualitative analysis. The quantitative analysis involved digital subtraction with each of the JPEG images to yield a new image using Photoshop 5.). The histograms of grey values were submitted to statistical analysis. The mean and standard deviation were calculated for each image. RESULTS: The data revealed that a JPEG lossy compression, six times smaller than the original TIFF, is compatible with diagnostic applications. CONCLUSIONS: The compression ratio calculated as the quotient between the file sizes and the standard deviation of the values corresponding to the image that resulted from digital subtraction may be employed to assess the outcome of the compression process and guarantee adequate quality. PMID- 12653316 TI - Effect of calcium hydroxide as a supplementary barrier in the radicular penetration of hydrogen peroxide during intracoronal bleaching in vitro. AB - AIM: To examine pH changes in the cervical external root surface, when calcium hydroxide was used as a supplementary barrier to the protective base material during intracoronal bleaching. METHODOLOGY: Twenty-eight single-rooted human premolars extracted for orthodontic reasons were instrumented with K-Flex files, obturated with gutta-percha and subjected to thermocatalytic bleaching. The teeth were divided into four groups. In group A, a glass-ionomer cement barrier was placed at the cemento-enamel junction (CEJ) level and in group C, the barrier was placed 1 mm apical to the CEJ. In groups B and D, Ca(OH)2 was placed in contact and apical to the glass-ionomer cement at the CEJ and 1 mm apical to the CEJ, respectively. The teeth were placed in vials containing distilled water and the pH values of the medium surrounding the teeth were recorded after 1, 2, 4, 10 and 15 days, following renewal of the medium. RESULTS: The pH in the medium became acidic in all groups. No statistically significant differences existed between groups for all the experimental days (P = 0.790). CONCLUSION: The placement of Ca(OH)2 as a supplementary barrier during intracoronal bleaching did not have a significant effect in reversing the acidic pH created at the external root surface in vitro. Its potential effect during these procedures in vivo needs to be further investigated. PMID- 12653318 TI - Location, arrangement and possible function of interodontoblastic collagen fibres in association with calcium hydroxide-induced hard tissue bridges. AB - AIM: To assess the location, arrangement and possible function of interodontoblastic collagen fibres in association with calcium hydroxide-induced hard tissue bridges by using light and transmission electron microscopy techniques and immunohistochemical staining localization. METHODOLOGY: Prior to the study, an animal use protocol form was reviewed and approved by the Screening Committee for Animal Research of the Tokyo Medical and Dental University. Exposed monkey pulps were capped with a hard-set calcium hydroxide and histopathologically evaluated at 3, 14, 21, 30 and 90 days, using light microscopy with silver staining and transmission electron microscopy to differentiate structural features of interodontoblastic collagen fibres. In addition, an attempt was made to identify and to differentiate between several types of collagen and fibronectin using immunohistochemical localization techniques. RESULTS: At 14 days, interodontoblastic collagen fibres were observed extending from the original dentine, passing through the odontoblasts, and consisted of two portions: a thick fibril and a thin fibril. At 21 days, interodontoblastic collagen fibres were seen penetrating into the predentine and becoming incorporated into the mineralized dentine. At 30 days, interodontoblastic collagen fibres reached the cell process. Although interodontoblastic collagen fibres were no longer observed near the odontoblastoid cells at the area of the newly formed tubular dentine, interodontoblastic collagen fibres were observed embedded within the primary formed dentine bridge. Immunohistochemical staining demonstrated type I collagen and fibronectin within the interodontoblastic collagen fibres. CONCLUSIONS: Interodontoblastic collagen fibres were routinely detected throughout early dentine bridges. Interodontoblastic collagen fibres are thought to be important for initial dentine bridging to induce and support a dentinogenesis framework. PMID- 12653319 TI - A comparative study of lateral condensation, heat-softened gutta-percha, and a modified master cone heat-softened backfilling technique. AB - AIM: The aim of the study was to compare the quality of root fillings completed by lateral condensation (L), Thermafil (TF), and a new technique using Thermafil for backfilling (BF), with special emphasis on control of overfilling. METHODOLOGY: Sixty curved canals in plastic blocks were prepared with the ProFile system to size 40/04 taper in the apical half and to 06 taper in the coronal half. The canals were divided into three groups of 20. Apical patency was verified with a size 15 K-file. The canals were then filled using three different techniques: LC (20 canals), TF (20 canals), and BF, where a size 40 master point with sealer was seated prior to the introduction of a size 30 Thermafil point (20 canals). The same resin-based sealer was used on each occasion. The root fillings were assessed using stereo-microscopy for material extrusion, digital radiography for occurrence of voids, and microscopy of sections for voids and thickness of sealer layer. RESULTS: Extrusion of both gutta-percha and sealer occurred in all 20 canals filled with the TF technique, but only three and five cases of sealer extrusion were detected with LC and BF techniques, respectively. No voids were detected in the TF group, whereas small voids were present in most fillings in the LC and BF groups. The average total length of the voids was less than 1 mm per canal. The thickness of the sealer layer in the middle and apical parts was greater in the LC and BF groups than in the TF group. CONCLUSIONS: LC and BF techniques resulted in fewer overfills than TF. Voids were absent in TF fillings, whilst small voids were found in the LC and BF groups. PMID- 12653320 TI - Prodromal herpes zoster--a diagnostic challenge in endodontics. AB - This case report highlights the diagnostic challenge that herpes zoster represents if pain develops in the prodromal stage. A 58-year-old male presented with pain in the left maxilla. The symptoms had lasted for 7 months. The first premolar had been extracted soon after the onset, and the second premolar root filled shortly thereafter. Symptoms were experienced as sudden pain attacks lasting for several hours and analgesics gave some pain relief. Clinical examination showed that the second premolar was tender to percussion. No sinus tract or swelling were present. Radiographic examination showed previously root filled second premolar and first molar teeth, and no evidence of apical pathosis. Due to the uncertainty about the quality of the root filling in the second premolar and incomplete root filling in the first molar, retreatment was started prior to prosthetic treatment in the region. The pain continued and became more intense during the treatment. A diagnosis of herpes zoster was determined, when an acute attack with oedema and vesicles occurred, 2 months after retreatment was started. In the present case, therefore, the primary attack presented itself after months and the herpes zoster diagnosis could not be made until then. points. Key learning points. * A long lasting prodromal stage is an unusual event. * Symptoms combined with inadequate technical standard of root fillings may confuse the diagnostics. * No available data support or suggest the use of antiviral treatment as a diagnostic tool. PMID- 12653321 TI - What is your diagnosis? Small intestinal mechanical obstruction. PMID- 12653322 TI - Owners' assessments of their dog's quality of life during palliative chemotherapy for lymphoma. AB - The owners of 25 dogs with multicentric lymphoma which were treated by multidrug chemotherapy were interviewed to assess the quality of their pet's life during treatment. Sixty-eight per cent of the owners considered their dog's quality of life to be the same as before the lymphoma occurred, and the remaining 32 per cent felt that their pet's quality of life on chemotherapy treatment was acceptable but poorer than before the lymphoma occurred. Treatment complications were reported by 52 per cent of owners. Ninety-two per cent of owners had no regrets about treating their dog with multidrug chemotherapy. The results suggest that, while many dogs developed treatment complications during multidrug chemotherapy, most owners felt that the quality of life of their pets was good during treatment and had no regrets about opting for palliative chemotherapy. PMID- 12653323 TI - Clinical validation of a proANP 31-67 fragment ELISA in the diagnosis of heart failure in the dog. AB - Atrial natriuretic peptide (ANP) is a polypeptide hormone found in increased concentrations in the plasma of dogs with heart failure. However, problems arise in using ANP as a diagnostic marker for heart failure because of its short half life in plasma, proteolysis post-collection and the necessity for a radioimmunoassay. The diagnostic utility of a proANP 31-67 ELISA for the detection of heart failure in dogs was evaluated using plasma collected from 31 dogs with clinical and radiographic signs of heart failure and control samples from 40 dogs considered to be free of cardiac disease. Log proANP 31-67 levels were significantly higher in the heart failure group (P < 0.001). In this population of dogs, using a cut-off value of 1,750 fmol/ml, the sensitivity and specificity of the assay were 83.9 per cent and 97.5 per cent, respectively. Using a cut-off of 1,350 fmol/ml, the sensitivity and specificity were 93.5 per cent and 72.5 per cent, respectively. It is concluded that a proANP 31-67 fragment ELISA should prove to be a useful diagnostic aid in naturally occurring canine heart failure. PMID- 12653324 TI - Healing of dorsal pancarpal arthrodesis in the dog. AB - The purpose of this study was to assess radiographically the healing process in dogs which had undergone dorsal pancarpal arthrodesis with autologous cancellous bone grafting. The medical records of 17 dogs that had undergone dorsal pancarpal arthrodesis with complication-free dynamic compression plate fixation were reviewed. Postoperative healing was evaluated radiographically. It started five weeks after surgery and progressed for six to 12 weeks in all dogs. After this time period no further progression of healing was observed. Between weeks 6 and 8, bony fusion progressed significantly more rapidly in the intercarpal joint than in the radiocarpal joint. The following criteria had no influence on healing time: gender, age, bodyweight, and immediate versus delayed surgery after trauma. The major difficulties noted with respect to the interpretation of postoperative radiographs were irregular bony bridging, mainly caused by inadequate palmar curettage, and oblique projection of articular surfaces resulting in superimposition of joint spaces and solid bone. PMID- 12653325 TI - Familial non-rcd1 generalised retinal degeneration in Irish setters. AB - Four Irish setters were diagnosed with bilateral retinal degeneration and cataracts at an age ranging from six to 11 years. In three of these dogs, progressive night blindness was reported from an age of eight to 11 years. In the fourth dog, aged six, no signs of visual impairment had been noticed. In all four dogs, the rod-cone dysplasia type 1 (rcd1) mutation was excluded as a cause, using an allele-specific PCR. From their three-generation pedigrees, a familial relationship was detected in three out of four dogs, which were also related to four additional Irish setter dogs with a history and clinical signs suggestive of late-onset progressive retinal degeneration. These results suggest the existence of a possibly hereditary, late-onset, progressive retinal atrophy in the Irish setter breed, that is distinct from rcd1. PMID- 12653326 TI - Septic fibrinous pericarditis in a cocker spaniel. AB - A four-year-old cocker spaniel presented with cardiac tamponade due to a pericardial effusion, in addition to pyrexia and peripheral neutrophilia and a recent history of chest trauma. Cytological examination of the pericardial effusion revealed a predominant neutrophilia. The echocardiographic findings were of numerous hyperechoic densities in the pericardial space, due to fibrin, with concurrent thickening and distortion of the pericardium. Postmortem examination, including microbiology, revealed the presence of organising septic fibrinous pericarditis associated with a mixed infection of Streptococcus canis, Citrobacter species, Pseudomonas species and alpha-haemolytic streptococci. PMID- 12653328 TI - Trilostane treatment of a dog with functional adrenocortical neoplasia. AB - A 13-year-old, crossbreed dog presented with a history of recent onset polydipsia, progressive lethargy, weakness and reduced appetite. Blood tests showed raised concentrations of alkaline phosphatase and alanine aminotransferase with marginally low serum potassium. There was a leucocytosis with a mature neutrophilia and no eosinophils. Endocrine tests showed a normal aldosterone concentration and an exaggerated adrenocorticotropic hormone (ACTH) stimulation test, consistent with a diagnosis of hyperadrenocorticism (HAC). A diagnosis of adrenal-dependent HAC was made, based on the presence of a calcified mass involving the left adrenal gland, and hepatomegaly, on radiography and ultrasonography. The owners declined surgical adrenalectomy. Medical management with trilostane rapidly improved the clinical signs and normalised the serum chemistry. ACTH stimulation tests showed an improvement in post-ACTH cortisol concentrations and were used to make dose adjustments where necessary. At the time of writing, no adverse side effects had been seen and the dog remained well after 80 weeks of treatment. PMID- 12653327 TI - Ocular manifestation of disseminated Mycobacterium simiae infection in a cat. AB - Disseminated mycobacterial disease was diagnosed in an eight-year-old domestic shorthaired cat, with involvement of the skin, lungs, lymph nodes and one eye. Mycobacterium simiae was cultured from skin biopsies on solid agar and in liquid media. This organism is known to cause pulmonary, cutaneous or disseminated infection in human patients with acquired immunodeficiency syndrome but has never been encountered as a pathogen in companion animals. Combination treatment with rifampicin, enrofloxacin and clarithromycin resulted in complete clinical remission within six months, with no side effects. No recurrence was observed in a 22-month follow-up period. PMID- 12653330 TI - Surgical treatment of right-sided renal lymphoma with invasion of the caudal vena cava. AB - An eight-year-old, male castrated basset hound presenting with a three-month history of lethargy was examined. Diagnostic tests including radiography and ultrasonography showed a right-sided renal mass. A 99mTc diethylenetriamine penta acetic acid scan demonstrated that this kidney was non-functional. At surgery, invasion of the caudal vena cava was found, and the renal segment of the vena cava and the right kidney were resected. The left renal vein was anastomosed to the more proximal vena cava using a polytetrafluoroethylene graft, and the dog recovered well. Two days postsurgery, the dog suffered an acute episode of aspiration pneumonia and was euthanased. The renal mass was diagnosed as lymphoma on histopathology. PMID- 12653329 TI - Dermatitis associated with Dirofilaria repens microfilariae in three dogs in Saudi Arabia. AB - The presence of microfilariae of the zoonotic nematode Dirofilaria repens is reported in three dogs from Saudi Arabia, manifesting as pruritic dermatitis with signs including erythema, alopecia, papules, crusting and hyperkeratosis. All dogs were found to have concurrent babesiosis. Specific treatments against the two conditions led to complete clinical recovery and disappearance of microfilariae from the blood one month later. PMID- 12653331 TI - Medulloblastoma in a cat: clinical and MRI findings. AB - A two-year-old, castrated crossbred cat presented with loss of balance and anorexia. A mass of the caudal aspect of the cerebellum was revealed by magnetic resonance imaging (MRI). The mass was hypointense on T1-weighted images, iso- and hyperintense on T2-weighted images and was enhanced by intravenous gadolinium contrast medium. The MRI characteristics of this case were similar to those of medulloblastoma of the cerebellar vermis in humans. The authors were able to remove almost all of the tumour. The cat was discharged from hospital on day 22 after surgery, but died on day 45. The excised tissue was histologically diagnosed as medulloblastoma. PMID- 12653332 TI - The assessment of an intra-oral video camera as an aid to occlusal caries detection. AB - AIMS: To investigate the use of intra-oral video cameras (IOVCs) for occlusal caries detection. DESIGN: Eighty premolars and molars set-up to simulate clinical conditions were examined by six examiners using three techniques: visual examination (EX), an IOVC (CAM) and by watching a video that was recorded using an intra-oral video camera (VID). The teeth were then sectioned for histological validation. RESULTS: 30 surfaces had lesions limited to enamel and 39 had lesions extending into dentine. Sensitivity scores at the D1 threshold (enamel and dentine lesions) were 43% (EX), 68% (CAM) and 77% (VID) whilst the specificity scores were 92% (EX), 64% (CAM) and 60% (VID). Corresponding results at the D3 threshold (caries extending into dentine) were 27%, 42%, 60%, 97%, 99% and 77%. CONCLUSIONS: Compared to unaided vision, the use of an IOVC significantly increases the number of occlusal lesions detected, however, there is a rise in false positive scores at the D1 threshold. The use of the video technique also significantly increases the number of occlusal lesions detected but with a concurrent rise in false positive scores at both thresholds. PMID- 12653334 TI - Malocclusion and orthodontic treatment need of handicapped individuals in South Canara, India. AB - AIMS: To estimate the prevalence and the severity of malocclusion and treatment need for 329 handicapped individuals. SUBJECTS AND SETTING: 329 handicapped individuals aged 11-30 years, attending eight different nonresidential special schools of South Canara, India. METHOD: Examination and recording using the Dental Aesthetic Index (DAI). RESULTS: 53% had a dental appearance which required no orthodontic treatment, 24% had a definite malocclusion, where treatment was 'elective' and treatment for a further 12% was considered to be 'highly desirable'. The remaining subjects (11%) had a handicapping malocclusion where treatment was considered mandatory. CONCLUSIONS: The prevalence of definite and severe malocclusions was higher in the mentally subnormal individuals when compared to other handicapping conditions. Orthodontic services for the handicapped have generally been neglected. PMID- 12653333 TI - Ethnic disparities in the prevalence of dental caries and restorative dental treatment in Brazilian children. AB - AIM: This study assesses disparities in the oral health status of Brazilian black and white children. PARTICIPANTS: 11- and 12-year-old schoolchildren living in 131 cities of the State of Sao Paulo, Brazil. METHODS: Spatial data analysis of city-level indexes of oral health, socio-economic status and provision of dental services. MAIN OUTCOME MEASURES: Ethnic ratios of the DMFT and the care index. RESULTS: White children had higher indexes of caries in permanent teeth than their black counterparts, concurrent with a higher utilisation of dental attendance. The gap of caries prevalence between black and white children was reduced in cities with a better profile of socio-economic status. Cities with higher per-capita yearly budget, expenditure in health, and provision of public dental services presented reduced indications of ethnic inequality in dental care. CONCLUSION: The knowledge of conditions associated with a lower ethnic discrepancy in the risk of caries and in the incorporation of dental services can be used to design socially appropriate dental services. An improved community dental service, higher public expenditure in health and per-capita municipal yearly budget contribute effectively to reducing inequities in oral health by allowing an incorporation of restorative dental treatment more equitably distributed between black and white children. PMID- 12653336 TI - A review of paediatric oral biopsies in Turkey. AB - OBJECTIVES: To determine the site and prevalence of oral lesions and the age and gender of a paediatric population in Turkey. DESIGN: A review of the archives of pathology departments. SETTINGS: Six pathology departments in Ankara, Turkey. METHODS: This study presents the findings of 472 oral biopsies from patients, up to 15 years of age, received over an eight-year period. The cases represent 5.5 per cent of the total number of reports examined (8,532). The data were revised and compiled for diagnosis site, age and sex. The patients were divided into three age groups according to dentition periods: primary dentition 0-5 years (Group 1), mixed dentition 6-12 years (Group 2), and permanent dentition 13-15 years (Group 3). The lesions were categorised into five major groups as: 1) reactive and inflammatory lesions 2) neoplastic lesions 3) cystic lesions 4) non neoplastic lesions 5) dermatologic lesions. RESULTS: Inflammatory and reactive lesions formed the largest group of diagnoses (49 per cent) followed by neoplasms (36 per cent) and cystic lesions (12 per cent). Approximately 60 per cent of the biopsies were from the second group patients with an almost equal distribution among sexes. The predominant site of the biopsies was gingiva (28 per cent) followed by lips (19.2 per cent). CONCLUSION: The majority (49 per cent) of the lesions were in the category of reactive and inflammatory lesions with most occurring in the second age group (6-12 years) that represents mixed dentition. The results of the study suggest that the difficulties in maintaining oral hygiene or the presence of trauma may be the primary factor in mucosal lesions occurring in the mixed dentition period. PMID- 12653335 TI - Guidance for the planning, implementation and evaluation of oral health care demonstration projects for under-served populations. AB - Recently, it has been proposed that in planning oral health care services in non established market economy (non-EME) countries, and for under- served communities throughout the world, high priority be given to a basic package of oral care (BPOC). This package contains three key components: emergency care (oral urgent treatment--OUT), exposure to appropriate fluoride (affordable fluoride toothpaste -AFT) and appropriate treatment technology (atraumatic restorative treatment- ART). These three components are embedded in the supporting context of oral health promotion (OHP). There is a lack of experience in implementing BPOC and besides, there is not much known about the effectiveness, efficiency and sustainability of the proposed components of BPOC, either individually or as a package, under local conditions. An effective approach in one setting may not be successful in another setting due to many factors such as a lack of acceptability by the community or local government or because of insufficient financial and human resources. It is therefore recommended to consider small scale demonstration projects for effectiveness, efficiency and sustainability assessments of the various components of BPOC before embarking on large scale programmes. The purpose of this paper is to highlight the different aspects related to the planning, implementation and evaluation of oral health demonstration projects for under-served communities. PMID- 12653338 TI - Case report: Oral pemphigus vulgaris with multiple oral polyps in a young patient. AB - A rare case of oral pemphigus vulgaris is presented, which developed initially in a 9-year old Jordanian male. The disease was not well controlled with immunosuppressive therapy and was complicated by the development of multiple oral polyps approximately one year after the onset of symptoms. These lesions were smooth, painless and located particularly on the upper labial gingiva, the labial and buccal mucosae and the tongue. Disease activity continued for approximately nine years until the patient was referred to an oral medicine specialist. Histological examination of the polyps revealed exuberant granulation tissue. Adjustment of the steroid dosage lead to resolution of the oral symptoms and the polyps reduced in number and in size. This case highlights the occurrence of pemphigus vulgaris in young patients, illustrates a rare complication of persistent ulcero-inflammatory disease and emphasises the importance of specialist referral in the management of oral disease. PMID- 12653337 TI - The comparison of wear characteristics of prosthodontic restorative materials. AB - AIM: To assess the mutual wear characteristics of various prosthodontic restorative materials. MATERIALS AND METHODS: Samples were prepared using base metal alloy, ceramics and aesthetic methyl metacrylate resin materials in cylinders with a diameter of 30 mm, height 4 mm, and a spherical shape with a diameter of 5 mm and length of 6 mm. The mutual wearing process of samples on each other in groups was tested using a pin-on disc wear tester machine. RESULTS: The data obtained were evaluated by means of variance analysis and T-test. It was determined that the percentages of the mutual wearing of the restorative materials were statistically significant (p<0.001), but whether the samples were in the upper or lower position had no statistical significance. CONCLUSION: Various prosthodontic restorative materials had wear characteristics affecting each other. PMID- 12653340 TI - A behavioural approach in the treatment of elderly patients: a new philosophy. AB - Complete denture prosthodontists are very well aware of the influence of psychological factors in the success of their treatment. Recently, a number of authors tried to ascertain some psychosocial criteria in the predictability of patients' satisfaction. Although special attention was paid to concerns of socio demographic and educational status, it was demonstrated that there is little relationship between quality of dentures and patients' satisfaction, nor a relationship between satisfaction and anatomical conditions. The purpose of this paper is to give a better understanding of the behavioural problems of this treatment and offer some solutions. Educational programmes should be influenced by behavioural problems in the management of the elderly patient. PMID- 12653339 TI - Late-life depression: its oral health significance. AB - Late-life depression (LLD) initially occurs after age 65 and is a major public health concern because elderly people who are at high risk constitute an ever expanding segment of the population. LLD is a mental illness in which mood, thought content, and behavioural patterns are impaired, causing individual distress, compromising social function and impairing self-maintenance skills (e.g. bathing, dressing, hygiene). It is characterised by marked sadness, or a loss of interest or pleasure in daily activities and may be accompanied by weight change, sleep disturbance, fatigue, difficulty concentrating, and high suicide rate. Individuals under treatment for LLD and those whose illness has not been diagnosed or treated often present to the dentist with significant oral disease. LLD is frequently associated with a disinterest in performing oral hygiene, a cariogenic diet, diminished salivary flow, rampant dental decay, advanced periodontal disease, and oral dysesthesias. Many medications used to treat the disease magnify the xerostomia and increase the incidence of dental disease. Appropriate dental management necessitates a vigorous preventive dental education programme, the use of artificial salivary products, antiseptic mouthwash, daily fluoride mouthrinse and special precautions when administering local anaesthetics with vasoconstrictors and prescribing analgesics. PMID- 12653341 TI - Daily dialysis in North America: evidence for a bright future. PMID- 12653343 TI - Effects of novel manufacturing technology on blood and dialysate flow distribution in a new low flux "alpha Polysulfone" hemodialyzer. AB - The main target for low flux hemodialyzers is an efficient low molecular weight solutes clearance. Such efficiency is largely dependent on the optimization of diffusion between blood and dialysis solution. The diffusion process can be impaired if there is a mismatch between blood and dialysate flow distribution in the dialyzer. Thus optimized flow distribution both in the blood and dialysate compartment becomes quintessential for the maximal efficiency of the diffusion process within the hemodialyzer. The present paper describes the distribution of the blood and dialysate flows in a new low flux polysulfone hollow fiber hemodialyzer characterized by a specific undulation of the fibers and a new cutting technology of the fibers for an improved micro-flow condition in the blood compartment headers. Twelve Diacap alpha Polysulfone LO PS 15 (1.5 sqm) (B. Braun Medizintechnologie, Melsungen Germany) were employed for the study. Six were analyzed in vitro and six were studied in vivo. Blood flow distribution was studied in vitro by dye injection in the blood compartment during experimental extracorporeal circulation utilizing human blood with hematocrit adjusted at 33%. Sequential images were obtained with a helical scanner in a fixed longitudinal section of the dialyzer 1 cm thick. Average and regional blood flow velocities were measured utilizing the reconstructed imaging sequence. The method allowed the calculation of single fiber blood flow (SF Qb) and the mass transfer zone (MTR) definition in digitally subtracted images. The patterns 20-10 and 40-30 were utilized. The same technology was used to evaluate flow distribution in the dialysate compartment after dye injection in the Hansen's connector. Regional dialysate flow was calculated in central and peripheral sample areas of 1 cm2. Six in vivo hemodialysis treatments on patients with end stage renal disease were performed at three different blood flow rates (250-350 and 450 ml/min) in order to measure urea, creatinine and phosphate clearance. Macroscopic and densitometrical analysis revealed that flow distribution was homogeneous in the blood compartment while in the dialysate compartment a slight difference between the peripheral and central regions in terms of flow velocity was observed. This however was not generating channeling phenomena. Urea creatinine and phosphate clearances were remarkably high and so were the Kt/V observed in all sessions, especially in relation to the studied blood flows. In conclusion, a significant blood to dialysate flow match with optimized countercurrent flow condition was observed in the studied hollow fiber hemodialyzers. Such optimization might be due both to the improved dialyzer design at the level of the blood header and to the specific fiber undulation that prevents dialysate channeling. PMID- 12653342 TI - Treatment of insulin resistance in uremia. AB - Insulin resistance is a characteristic feature of uremia. As long as the hyperinsulinemia adequate to overcome the insulin resistance, glucose tolerance remains normal. In patients destined to develop type 2 diabetes, the beta cell compensatory response declines, and relative, or absolute, insulin deficiency develops. At this point glucose intolerance and eventually frank type 2 diabetes occur. Insulin resistance and concomitant hyperinsulinemia are present irrespective of the type of renal disease. Several studies have confirmed that hemodialysis (HD) treatment significantly improves insulin resistance. Both CAPD and CCPD are shown to improve insulin resistance in uremic patients. Comparing the effect of PD and HD treatment, it was found that the CCPD group has significantly higher insulin sensitivity than the HD group with the CAPD group similar to HD. Treatment of calcium and phosphate disturbances, including vitamin D therapy, significantly reduces insulin resistance in uremia. Treatment with recombinant human erythropoietin (EPO) is an efficient way to increase hematocrit, to reverse cardiovascular problems and to improve insulin sensitivity. Angiotensin-converting enzyme inhibitors have been shown to improve insulin resistance, hyperinsulinemia and glucose intolerance in uremic patients. Thiazolidinediones (TZDs), the new insulin-sensitizing drugs, provide the proof that pharmacologic treatment of insulin resistance can be of enormous clinical benefit. The great potential of insulin resistance therapy illuminated by the TZDs will continue to catalyze research in this area directed toward the discovery of new insulin-sensitizing agents that work through other mechanisms. PMID- 12653344 TI - Impact of blood and dialysate flow and surface on performance of new polysulfone hemodialysis dialyzers. AB - Optimization of hemodialysis treatment parameters and the characteristics of the dialyzer are crucial for short- and long-term outcome of end stage renal disease patients. The new high-flux membrane Helixone in the dialyzer of the FX series (Fresenius Medical Care, Germany) has interesting features, such as the relationship of membrane thickness and capillary diameter which increases middle molecule elimination by convection, as well as higher capillary packing and microondulation to improve the dialysate flow and distribution. Blood flow, dialysate flow and surface area are the main determinants of the performance of a dialyzer, however the impact of each parameter on small and middle molecule clearance in high flux dialysis has not been well explored. In order to find the best treatment condition for the new dialyzer series, we evaluated urea, creatinine, phosphate clearances and reduction rate of beta2-microglobulin in ten stable patients treated with different blood flows (effective Qb 280 and 360 ml/min), dialysate flow (Qd 300 or 500 ml/min) and dialyzer surfaces (1.4 and 2.2 m2, FX60 or FX100). KoA and Kt/V were also calculated. Blood flow, dialysate flow and surface area demonstrated a significant and independent effect on clearance of urea, creatinine and phosphate, as well as on Kt/V. Small solute clearance was stable over the treatment. In contrast to small solutes, reduction rate of beta2 microglobulin was related to increasing dialyzer surface only. The new dialyzer design of the FX series proves highly effective due to improved dialysate distribution and reduced diffusive resistance as shown by the small solute clearance. A high reduction rate of beta2-microglobulin is favored by improved fiber geometry and pore size distribution. These findings have potential long term benefits for the patient. PMID- 12653345 TI - Peritoneal leukocyte survival and respiratory burst responses in patients treated with a low glucose degradation and high pH peritoneal dialysis fluid. AB - The discovery of toxicity related to glucose degradation products (GDP) has initiated the development of new PD fluids with low GDP concentrations and higher, more physiological, pH levels. Cell numbers, differential counts and the respiratory burst responses of peritoneal leukocytes were compared between patients treated with the low GDP, high pH fluid Gambrosol-trio (n=10) and a conventional fluid (n=12). Effluents from over-night dwells were collected and leukocytes were evaluated morphologically and by luminol-amplified chemiluminescence (CL) after stimulation with opsonized zymosan. The frequency of necrosis and early apoptosis was quantified by means of annexin V binding and propidium iodide uptake. The Gambrosol-trio group produced significantly higher (p<5%) macrophage counts and stronger CL responses (p<10%) than did the conventional fluid group. The cell compositions did not differ significantly between the groups. Necrosis was significantly more common among the cells in the conventional fluid group. The occurrence of apoptosis did not differ between the fluids. PMID- 12653347 TI - Pre-HD dilution acidosis, without post-HD contraction alkalosis in uremic patients. AB - The aim of this study was to verify if the degree of pre-HD acidosis and its correction post-HD is related to body fluid expansion during the interdialytic period. Twelve uremic patients without major problems, with stable hematocrit, with regular and similar HD-session characteristics, but widely varying amounts of body fluid expansion in the interdialytic period were included. Blood samples were collected from arterial line pre- and post-HD, anaerobically in heparinized syringes, for determination of HCO3-, pH and PaCO2 (radiometer Copenhagen ABL 300 Acid-Base Laboratory), in two similar HD-sessions for each patient (12 patients, 24 HD-sessions). The percentage (%) of body weight gain in the interdialytic period was also estimated. For each patient, the mean value of parameters studied in the two HD-sessions was used for the evaluation of findings. According to mean values (+/-SD) of HCO3-, pH and PaCO2 Pre-HD (18.26+/-1.99 mmol/L, 7.31+/-0.03, 36.27+/-2.5 mmHg respectively) and post-HD (26.37+/-1.7, 7.43+/-0.03, 38.43+/ 2.10 respectively) patients are acidotic pre-HD and slightly alkalemic post-HD. Correlation between the percentage (%) of interdialytic body weight gain (IBWG) and the values of HCO3-, pH and PaCO2, Pre-HD (r=-0.814, p<0.001; r=-0.931, p<0.001; r=0, 100 NS; respectively) and post-HD (r=-0.958, p<0.001; r=-0.937, p<0.001; r=-0.504 NS; respectively) indicates a significant and negative relationship of IBWG% with HCO3- and pH pre- and post-HD, but not with PCO2. In conclusion, the negative relationship of IBWG% with HCO3- and pH pre- and post-HD indicates that the body fluid expansion during the interdialytic period contributes to a dilutional acidosis pre-HD, but not to a contraction alkalosis post-HD, by the elimination of fluid during the HD-session. PMID- 12653346 TI - Effect of hollow fiber length on solute removal and quantification of internal filtration rate by Doppler ultrasound. AB - Renal replacement therapy with dialyzers capable of enhanced internal filtration (IF) can be an alternative to standard hemodiafiltration, as it provides convective solute removal comparable to that of hemodiafiltration by a simple procedure. In this study, we clinically evaluated the effect of the hollow fiber length in the dialyzer, a crucial factor influencing the rate of IF, by comparing two commercial dialyzers (BS-1.6U, BS-1.6UL, Toray, Japan) which differed in the fiber length, but had the same surface area and inner diameter of their hollow fibers. We showed that in the dialyzer with the longer fibers, the pressure profile along the dialyzer was significantly altered, and the solute clearance tended to be increased. In addition, we successfully quantified the IF rate with a Doppler ultrasound in the experimental circuit, by measuring the blood flow velocities along the bundle of fibers. We showed that the changes in the blood flow velocity were more marked in the dialyzer with the longer fibers; the calculated IF rates in the dialyzers with the shorter and longer fibers were 11.1 mL/min and 37.7 mL/min, respectively, which seemed to be compatible with the solute clearances. This simple and readily applicable method is expected to be useful in the development of modified dialyzers to fully exploit the benefits of IF in renal replacement therapy. PMID- 12653349 TI - Hepatic encephalopathy and cerebral blood flow improved by liver dialysis treatment. AB - Eight acute liver failure patients, all in grade IV hepatic encephalopathy, were administered liver dialysis treatment with the Hemo Therapies Unit (Hemo Therapies Inc, San Diego, CA, USA). The patients were evaluated to determine whether the Glasgow Coma Scale score and cerebral blood flow improved with treatment. After the initial treatment, consciousness levels as measured by the Glasgow Coma Scale improved from a pre-treatment median of 5 (range 3 to 6) to a post-treatment median of 7 (range 5 to 9) (p=0.0005 by paired Wilcoxon test); mean blood flow velocity in the middle cerebral arteries as shown by transcranial Doppler sonography increased from a median of 37.85 cm/sec (range 20.3 to 114.0) to 57.90 (32.5 to 135.0) post-treatment (p=0.022); however, there was no significant change in the pulsatility index from a median of 1.18 (range 0.61 to 1.71) to 0.85 (range 0.70 to approximately 1.79) post-treatment (p=0.13). The 8 patients received 2 to 7 (median 5.5) times of daily 6-h liver dialysis treatments. Following the completion of all liver dialysis treatments, hepatic coma was fully resolved in 4 of 8 patients (50%) Three of 8 patients (37.5%) survived to hospital discharge, whereas 5 patients did not survive due to irreversible liver function and associated complications. In conclusion, liver dialysis treatment could improve hepatic encephalopathy, but the prognosis still depended on the underlying diseases. PMID- 12653348 TI - Modulation of pro-apoptotic (Bax) and anti-apoptotic (Bcl-2) gene expression in isolated porcine hepatocytes perfused within a radial-flow bioreactor after low temperature storing. AB - Due to the scarcity of available human livers, porcine hepatocytes are currently being evaluated as a xenogeneic cell source for extracorporeal bioartificial liver (BAL). Hypothermic storage of isolated porcine hepatocytes could support stocking of cell-loaded bioreactors for BAL use and may provide bioreactors ready to be used at the patient's bedside. For the development of this technology, it is of utmost importance to ensure cell viability and differentiated functions after low-temperature storage and following warm reperfusion. We compared cell viability, functional activity and apoptosis in isolated porcine hepatocytes which were perfused within a radial-flow bioreactor (RFB), stored at 4 degrees C and then reperfused at 37 degrees C. RFBs were loaded with 8 x 10(9), > or = 90% viable hepatocytes at 37 degrees C for 3 h. RFBs were then flushed with 4 degrees C University of Wisconsin solution (UW) and subsequently stored for 24 h or 48 h. RFBs were then reperfused for 8 h with recirculating medium plus serum at 37 degrees C . Cytochrome P450 (CYP) activity was studied before and after cold storage by means of monoethylglycinexylide (MEGX) detection in the effluent medium, after repeated lidocaine injections. After reperfusion experiments, hepatocytes were harvested for total RNA isolation. Reverse transcriptase polymerase chain reaction (RT-PCR) was used in order to amplify specific mRNAs for Bcl-2 and Bax genes, by using appropriate primers; beta-actin primers were used as control. Total RNA was extracted by northern blotting analysis and for Bcl-2, Bax and beta-actin RNA messenger detection, RT-PCR amplification was used. Freshly isolated hepatocytes perfused into the RFB showed a progressive increase of MEGX while a loss in Bax expression was paralleled by an increase in Bcl-2 expression, in comparison to starting hepatocytes. After 4 degrees C storage and warm reperfusion, MEGX production was preserved in 24 h- and 48 h-stored bioreactors as well as a sharp increase of Bcl-2 and a decrease of Bax mRNAs. Our study suggests that refrigeration of hepatocyte-bioreactors is a suitable strategy to maintain both viability and function of isolated hepatocytes, for up to 48 h a time-length that is compatible with long-distance delivery of ready-to use bioreactors. PMID- 12653350 TI - Numerical simulation of carotid hemodynamics in patients with rotary blood pump cardiac assist. AB - In recipients of rotary blood pumps for cardiac assist, the pulsatility of arterial flow is considerably diminished. This influences the shear stress patterns and streamlines in the arterial bed, with potential influence on washout and subsequent plaque growth. To study these effects, a three-dimensional computer simulation of the carotid bifurcation at various levels of flow pulsatility was performed. The results showed that as expected pulsatile shear stress varied considerably, whereas local mean shear stress levels were nearly identical for all degrees of pulsatility. Particle residence time in the carotid bulb did only increase for less than 15%, with secondary washout patterns contributing to good washout also in nonpulsatile conditions. It is concluded that also under continuous pump support the local flow patterns in the carotids provide sufficient washout and fluid exchange to prevent excessive plaque growth. PMID- 12653353 TI - Survival and function of Tenckhoff peritoneal dialysis catheter after surgical or percutaneous placement: one centre experience. PMID- 12653351 TI - Delivery of high dose VEGF plasmid using fibrin carrier does not influence its angiogenic potency. AB - Delivery of DNA mixed with a degradable matrix carrier was supposed to improve transgene expression. Using a rabbit hind-limb ischemia model, we tested the angiogenic potency of plasmid encoding human vascular endothelial growth factor (pSG5-VEGF165) entrapped in fibrin sealant. Animals were injected intramuscularly with 500 microg of pSG5-VEGF165 or control plasmid, dissolved in saline (PBS) or fibrin glue. After 14 days, presence of delivered constructs and expression of transgene was confirmed in injected muscles of all animals. There were no significant differences in the levels of human VEGF mRNA and protein between VEGF PBS and VEGF-fibrin groups (Mann-Whitney test). Accordingly, pSG5-VEGF165 regardless of the way of delivery, induced similar increases in capillary density within treated muscles (ANOVA). Control plasmid did not show any effects. In conclusion, injection of pSG5-VEGF165 into ischemic adductor muscle leads to synthesis of human VEGF and increases the number of capillaries. Fibrin carrier does not influence its angiogenic potential. PMID- 12653352 TI - Plasmaexchange in the treatment of myasthenia gravis associated with thymoma. AB - BACKGROUND: All patients with thymomatous Myasthenia Gravis (MG) should undergo early and total thymectomy, but controversy abounds in the choice of chronic immunosuppressive agents. The value of plasmaexchange (PE) in MG has been clearly established in preoperative preparation and treatment of myasthenic crisis. Whether PE may be used in the chronic long-term therapy of patients with thymomatous MG in addition to conventional immunosuppressive agents and cholinesterase inhibitors is yet to be answered. CASE HISTORY: We present a 40 year old woman with an 11 year history of MG. Thymectomy was done during the first year of the disease and the histopathologic finding was thymoma. To sustain clinical remission after thymectomy she continued with immunosuppression with methylprednisolone and cyclosporin A (or azathioprine) in addition to cholinesterase inhibitors. Despite the almost continuous immunosuppression, the disease course continued with fluctuating myasthenic weakness which few times progressed to myasthenic crisis requiring mechanical ventilation. During myasthenic crisis we performed 6-8 plasmapheresis at 2-3 day intervals in addition to conventional immunosuppressive therapy. The disease rapidly worsened in January 2000 and we started with intermittent plasmapheresis (3-6 procedures at 2-3 day intervals, every 6-8 weeks) in order to sustain remission. With this therapeutic protocol, during 20 months follow-up we managed to prevent myasthenic crisis and to avoid ventilatory support. CONCLUSIONS: Plasmaexchange could be used as a successful and safe therapeutic tool in chronic long-term therapy in addition to conventional immunosuppressive agents to sustain remission in patients with MG. This is particularly important in the treatment of patients with thymomatous MG because they have an increased frequency of myasthenic crisis and often respond poorly an to immunosuppression with steroids or other immunosuppressants. PMID- 12653355 TI - Statistics of polarization-dependent gain in fiber-based Raman amplifiers. AB - We develop an analytic model for finding the statistics of polarization-dependent gain (PDG) in fiber-based Raman amplifiers. We use it to find an analytic form for the probability distribution of PDG and study how the mean PDG and the variance of PDG fluctuations depend on the PMD parameter. We show that mean PDG as well as PDG fluctuations are reduced by approximately a factor of 30 in the case of backward pumping. PMID- 12653354 TI - Enhancement of Raman scattering by deformation of microparticles. AB - Raman scattering on deformed droplets levitated in an acoustic levitator and produced by a vibrating-orifice aerosol generator were investigated. Our samples experiments were diethyl hexyl sebecate (DEHS) droplets in the millimeter-size range and ethanol droplets in the size range 50-100 microm. The C-H stretching region from 2800 to 3100 cm(-1) was investigated. We found that the Raman intensity measured by a scattering angle of 90 degrees depended on the shape of the droplets. Raman scattering on spherical droplets was smaller than scattering on spheroidal droplets with the same volume. Similar results were observed for the fluorescence signal of Rhodamine 6G-doped DEHS droplets. PMID- 12653356 TI - Bragg gratings in air-silica structured fibers. AB - We report on grating writing in air-silica structured optical fibers with pure silica cores by use of two-photon absorption at 193 nm. A decrease in propagation loss with irradiation was observed. The characteristic growth curves were obtained. PMID- 12653357 TI - Investigation of microdeformation-induced attenuation spectra in a photonic crystal fiber. AB - We investigate both theoretically and experimentally the induced spectral attenuation in an all-silica photonic crystal fiber subjected to periodic axial microdeformations. The induced attenuation spectra show discrete attenuation peaks with a spectral position that is dependent on the period of the induced deformation. The peaks are assumed to be the result of mode coupling between the fundamental mode and a highly lossy higher-order mode. This assumption is verified through numerical calculation of the beat length between these two modes. Excellent agreement between experiment and numerical predictions of the spectral position of the attenuation peaks is obtained. PMID- 12653359 TI - Improvement of the spatial beam quality of laser sources with an intracavity Bragg grating. AB - We demonstrate a novel method for improvement of spatial beam quality by use of the angular selectivity of a thick phase grating inserted into various laser cavities. Preliminary experiments with an intentionally degraded beam from a Nd:YVO4 laser have enhanced beam propagation factor M2 by a factor of 4.5. Operation with a broad-area laser diode in an external cavity with a thick grating was achieved. We obtained 100 mW of cw power in a beam with an M2 of 1.3. Moreover, a reduction of the output spectrum by as much as a factor of 3.8 was obtained. PMID- 12653358 TI - Fluorescence quantum efficiency measurements in the presence of Auger upconversion by the thermal lens method. AB - Mode-mismatched thermal lens (TL) measurements in Cr3+-doped fluoride crystals (LiSrAIF6 and LiSrGaF6) are reported. A nonlinear increase of the TL signal, and decrease of quantum efficiency, with increasing excitation power was observed and attributed to energy-transfer upconversion (ETU). Assuming an upconversion rate that is proportional to the excited-state population, Wup = gammaN(e), the theoretical model developed fits the experimental data well. The ETU parameters (gamma) were determined with unprecedented accuracy for low Cr concentrations. Thermo-optical parameters (K, ds/dT) were also obtained. PMID- 12653360 TI - Frequency-stabilized high-power violet laser diode with an ytterbium hollow cathode lamp. AB - We have demonstrated in an ytterbium laser cooling and trapping experiment a high power violet extendedcavity diode laser (ECDL) stabilized to the Yb resonant transition at 398.9 nm in an Yb hollow-cathode lamp. A frequency-dispersion signal, which we obtained by applying a modulation-free dichroic-atomic-vapor laser lock technique, allowed us to stabilize the violet ECDL at a frequency stability below 1 MHz at 1-s average time and a useful output power of 15 mW. PMID- 12653361 TI - 2.1-microm lasers frequency stabilized against CO2 lines: comparison between fringe-side and frequency-modulation locking methods. AB - A comparison between the use of the fringe-side and the frequency-modulation methods to frequency stabilize diode-pumped Tm-Ho:YAG lasers against linear absorption lines of CO2 near 2.1 microm is reported. Frequency stability was evaluated by monitoring of the beat signal between two independently stabilized Tm-Ho:YAG systems. Frequency instability levels of approximately 20 and approximately 400 kHz for 1-s integration time were achieved by the frequency modulation and fringe-side methods, respectively. PMID- 12653362 TI - Stable spatial solitons in semiconductor optical amplifiers. AB - The existence of stable dissipative spatial solitons at low intensities in patterned electrode semiconductor optical amplifiers (SOAs) is predicted theoretically. In contrast to conventional SOAs, this system may support stable solitons because the inherent saturating losses provide subcritical bifurcations for both the plane-wave and the soliton solution. PMID- 12653363 TI - Generation of vacuum-ultraviolet light by an optically contacted, prism-coupled KBe2BO3F2 crystal. AB - We have demonstrated the second harmonic of a frequency-tripled Nd:YVO4 laser with 2.5-mW quasi-cw output by using an optically contacted, prism-coupled KBe2BO3F2 crystal. We also achieved the second harmonic with a frequency-doubled single-mode Ti:sapphire laser at 172.5 nm and sum-frequency mixing with a dual wavelength Ti:sapphire laser at 163.3 nm. These wavelengths are to our knowledge the shortest obtained by use of nonlinear crystals for second-harmonic generation and sum-frequency mixing, respectively. PMID- 12653364 TI - High-power and high-contrast optical parametric chirped pulse amplification in beta-BaB2O4 crystal. AB - We have developed a high-power laser system based on optical parametric chirped pulse amplification in a beta-BaB2O4 crystal. The system provides gain of 10(8), a conversion efficiency of approximately 23%, an output energy of -65 mJ, and good amplified beam quality. The prepulse ratio has been measured to be 1.5 x 10( 8) or less. The spectral width was as broad as 16.5 nm centered at 1053 nm. PMID- 12653365 TI - Emergence of linear wave segments and predictable traits in saturated nonlinear media. AB - We find the key behind the existence traits of asymptotic saturated nonlinear optical solitons in the emergence of linear wave segments. These traits, produced by the progressive relegation of nonlinear dynamics to wave tails, allow a direct and versatile analytical prediction of self-trapping existence conditions and simple soliton scaling laws, which we confirm experimentally in saturated-Kerr self-trapping observed in photorefractives. This approach provides the means to correctly evaluate beam tails in the saturated regime, which is instrumental in the prediction of soliton interaction forces. PMID- 12653366 TI - Efficient 355-nm generation in CsB3O5 crystal. AB - We demonstrate UV generation in CSB3O5 (CBO) crystals grown by the top-seeded solution growth technique. 355-nm UV light was generated by use of a type II CBO crystal as a sum frequency of the fundamental light and the second harmonic of a nanosecond Nd:YVO4 laser. A 3.0-W output of the third-harmonic was obtained at a repetition rate of 31 kHz. The conversion efficiency from the fundamental light to the third harmonic reached 30%, which was 1.5 times higher than that obtained with a type II LiB3O5 crystal under the same experimental conditions. PMID- 12653367 TI - Nonlinear photoluminescence from van Hove singularities in multiwalled carbon nanotubes. AB - Van Hove singularities (VHS) in the density of states of multiwalled carbon nanotubes (MWNTs) have been observed by optical spectroscopy for the first time to the authors' knowledge. Additionally, unexpected visible luminescence has been observed on photoexcitation at 1064 nm. This luminescence is believed to arise from radiative transitions between energy states in the VHS. A mechanism describing the underlying processes, which involve multiphoton absorption followed by upconverted luminescence from VHS energy states, is presented. Our results show that the presence of VHS readily facilitates efficient optical transitions in PMID- 12653368 TI - Optical image encryption by random shifting in fractional Fourier domains. AB - A number of methods have recently been proposed in the literature for the encryption of two-dimensional information by use of optical systems based on the fractional Fourier transform. Typically, these methods require random phase screen keys for decrypting the data, which must be stored at the receiver and must be carefully aligned with the received encrypted data. A new technique based on a random shifting, or jigsaw, algorithm is proposed. This method does not require the use of phase keys. The image is encrypted by juxtaposition of sections of the image in fractional Fourier domains. The new method has been compared with existing methods and shows comparable or superior robustness to blind decryption. Optical implementation is discussed, and the sensitivity of the various encryption keys to blind decryption is examined. PMID- 12653369 TI - Shift of whispering-gallery modes in microspheres by protein adsorption. AB - Biosensors based on the shift of whispering-gallery modes in microspheres accompanying protein adsorption are described by use of a perturbation theory. For random spatial adsorption, theory predicts that the shift should be inversely proportional to microsphere radius R and proportional to protein surface density and excess polarizability. Measurements are found to be consistent with the theory, and the correspondence enables the average surface area occupied by a single protein to be estimated. These results are consistent with crystallographic data for bovine serum albumin. The theoretical shift for adsorption of a single protein is found to be extremely sensitive to the target region, with adsorption in the most sensitive region varying as 1/R(5/2). Specific parameters for single protein or virus particle detection are predicted. PMID- 12653370 TI - Generation of entanglement between frequency bands by nonlinear fiber propagation and spectral pulse shaping. AB - A novel fiber-based scheme for generating quadrature entanglement between a specific pair of frequency bands is proposed. The scheme is based on use of a nonlinear fiber, a spectral pulse shaper, and an adaptive feedback loop. It is numerically predicted that at least -5 dB of the quadrature entanglement will be created by preparation of an appropriate initial phase spectrum. Optimal control of the four-wave mixing terms of the Kerr Hamiltonian is crucial for improvement of the entanglement. PMID- 12653371 TI - Single-shot kilohertz characterization of ultrashort pulses by spectral phase interferometry for direct electric-field reconstruction. AB - We describe a method of characterizing ultrashort optical pulses that is based on the technique of spectral phase interferometry for direct electric-field reconstruction and is capable of simultaneously measuring the amplitude and the phase of the electric field of a sub-10-fs pulse at kilohertz acquisition rates on a single-shot basis. Use of this technique results in a dramatic increase (>50x) in acquisition rate compared with that of existing diagnostics for full E field characterization and opens the door to a range of new experiments in which shot-to-shot phase and amplitude fluctuations are studied at kilohertz rates. PMID- 12653372 TI - Amplification and compression of ultrashort fundamental solitons in an erbium doped nonlinear amplifying fiber loop mirror. AB - A nonlinear amplifying loop mirror constructed from erbium-doped fiber is proposed for simultaneous amplification and compression of ultrashort fundamental solitons. Numerical simulations show that, unlike conventional erbium-doped fiber amplifiers in which nonlinear effects lead to serious degradation of pulse quality, the proposed device performs efficient high-quality amplification and compression of ultrashort fundamental solitons while it almost completely preserves the soliton nature of the input pulses. Moreover, the performance of the device is insensitive to small variations in the loop length and in the power splitting ratio of the coupler. PMID- 12653373 TI - Misadventures in insulin therapy: are you at risk? AB - About dollar 1 out of every dollar 7 spent on health care is related to diabetes mellitus, a leading cause of blindness and kidney failure and a strong risk factor for heart disease. Prevalence of the disease has increased by a third among adults in general in the last decade, but intensive therapy has been shown to delay the onset and slow the progression of diabetes-related complications. While insulin therapy remains key in the management of type 1 diabetes, many patients with type 2, or insulin-resistant, diabetes encounter insulin administration errors that compromise the quality of insulin delivery. Insulin errors are a major, but modifiable, barrier to dosing accuracy and optimal diabetes control for many patients. Future trends to combat the problem include increased use of insulin inhalers and smaller doses of rapid- or short-acting insulin to supplement longer-acting injections. PMID- 12653374 TI - The "slave health deficit:" the case for reparations to bring health parity to African Americans. PMID- 12653375 TI - Teaching patients to communicate with physicians: the impact of race. AB - Research on physician-patient communication has focused on the effect of physician communication training on health care outcomes. Much less is known about patient communication training, and even less about the impact of patient race on the effectiveness of patient communication interventions. One hundred and fifty patients of 25 family physicians were randomly assigned to one of three groups: 14-page patient communication workbook received 2-3 days pre-visit, 2 page patient communication handout received in the waiting room, or control group. Racial differences in the impact of patient communication training communication variables, immediate and delayed recall of information, and adherence to treatment were analyzed by t-test and ANOVA techniques. Across analyses, workbook communication skills training had a strong and significant effect on white patients but minimal or no effect on African-American patients. Minimal differences in dependent variables between racial groups existed for the patient handout and control groups. A partial correlation analysis was conducted to factor out the variance due to education. Results suggested that patient race accounted for the results over and above differences in education between racial groups. Our results suggested that the benefits of communication training can be enhanced by taking into account patient characteristics such as race and culture. PMID- 12653376 TI - Knowledge, beliefs, and risk factors for osteoporosis among African-American and Hispanic women. AB - The purpose of this work was to develop and conduct a needs and risk instrument to assess knowledge of osteoporosis risk factors, identify beliefs and attitudes about this disease, and delineate the presence and/or absence of healthy behaviors associated with osteoporosis among African American and Hispanic women. The survey findings suggest that African-American and Hispanic women are not well versed in behaviors that would promote and maintain optimal bone mass. Consequently, they are not practicing appropriate lifestyle and dietary habits to decrease their risk of osteoporosis. Such behaviors include inadequate physical activity, inadequate calcium intake, cigarette smoking, and long-term steroid use. Less than 10% of women in the study were getting adequate daily dietary calcium intake, with only 13% taking daily calcium supplements to augment this deficit and less than one-half of women exercising at a minimal level (20 minutes/3 times a week). Women in this study also had limited knowledge about osteoporosis, perceived this condition to be less of a health threat as compared to breast cancer, heart disease, diabetes, and Alzheimer's disease, and very few had the perception that being Hispanic or African American was a factor to consider in assessing their risk of osteoporosis. Our findings suggest that osteoporosis education and prevention initiatives are needed, specifically for African-American and Hispanic women, to promote healthy behaviors, identify women at-risk, and encourage early diagnosis and treatment. PMID- 12653377 TI - Adult constipation: a review and clinical guide. AB - Constipation is a common complaint that can be a symptom of serious disease. Awareness of the potential etiologies can help direct the history, physical exam and subsequent work-up for the presenting individual. This article details the differential diagnosis and pathophysiology of constipation based on a review of the literature. The article is also designed to be useful as a guide to the work up of constipation. Key elements of the history, physical exam and testing are outlined. Included is a detailed flow diagram to guide the work-up of constipation. Testing methods and their value in the evaluation of chronic idiopathic constipation are discussed. Finally, although the focus of this article is the evaluation of constipation, a section on the treatment of constipation is included. PMID- 12653378 TI - Delayed diagnosis of cluster headache in African-American women. AB - The male-to-female ratio has fallen in cluster headache over the last several decades and is now 2.1:1. Unfortunately, women still are not diagnosed accurately. This lack of appropriate diagnosis appears related to the misconception that cluster headache rarely occurs in women. Compounding this misconception, there seems to be an ethnic bias. We report cluster headache in five African-American women in whom diagnosis was delayed due to gender, ethnicity, and, most importantly, an inability to make a correct diagnosis of cluster headache. Cluster headache diagnostic criteria are no different in men or women and have no ethnic boundaries. Clinical features such as disordered chronobiology and abnormal behavior often suggest the diagnosis. Migrainous features occur commonly in cluster headache and, when present, should not exclude the diagnosis. Likewise, neither race nor sex should exclude the diagnosis. The diagnosis of cluster headache is easily made by considering unilateral orbital, supraorbital or temporal location; short duration (15-180 minutes, untreated), and ipsilateral autonomic dysfunction involving the eye or nose. PMID- 12653379 TI - Adrenal cortical carcinoma: a case of late pulmonary metastasis. AB - A 71 year-old woman with a large tumor of the upper lobe of the left lung was diagnosed as having metastatic adrenal cortical carcinoma nine years after the initial adrenalo-nephrectomy. The patient was well with no recurrence for six years following resection of the single lung metastasis. PMID- 12653380 TI - Breast cancer and AIDS. PMID- 12653381 TI - Factors preventing African Americans from seeking early intervention in the treatment of ischemic strokes. AB - The most widely advocated drug that has been developed to decrease and possibly reverse the crippling effect of stroke is recombinant tissue plasminogen activator (rt-PA). In the treatment of ischemic stroke, rt-PA must be administered within the first 3 hr after the onset of symptoms for optimal efficacy and avoidance of life-threatening cerebral hemorrhage. This study sought to determine whether African-American stroke victims presented for treatment within the first 3 hr. In the study, we identified some of the more common reasons for the lack of prompt presentation. A prospective survey of 103 randomly selected acute stroke patients was performed. We found that less than half of this population sought assistance in the emergency department in time to take advantage of rt-PA therapy. When confronted with symptoms that were consistent with acute stroke, many patients either chose to ignore them or thought that they were experiencing transient phenomena. Three factors were identified as possible reasons for the lack of prompt response: (a) a lack of information about available treatment options; (b) the inability to recognize the early warning signs of an impending stroke; and (c) denial of illness. This study may help to evolve strategies that must be instituted to better educate the community about the early recognition of stroke and available treatment alternatives. PMID- 12653382 TI - Perceived needs of African-American caregivers of elders with dementia. AB - To assess the perceived needs of African-American caregivers and their expectations of the health care system, perceived level of success and satisfaction in meeting their needs, and their level of distress, we convened two focus groups at a local church in Oklahoma City, OK to discuss three questions. Responses were qualitatively analyzed for common themes. The groups comprised 13 African-American caregivers of demented elders. All participants completed a demographic form, the Zarit Burden Interview, and the Depression Scale. We found that African-American caregivers are frustrated with health care and service providers. The main differences of this group compared with other caregivers were their perceptions of racism and the absence of an available local support group. This information can serve to guide future studies in developing ways to increase caregiver satisfaction, decrease caregiver stress, and increase patient quality of life for this subgroup of caregivers. PMID- 12653383 TI - Sickle cell crisis in the adult: chest radiographic findings and comparison with pediatric sickle cell disease. AB - With the advent of improved therapy, an increasing proportion of individuals suffering from sickle cell disease (SCD) are surviving into adulthood. In contrast to children, little has been documented concerning the typical radiographic findings in adults presenting with sickle cell crises (SCC). We describe the chest radiographic (CXR) manifestations of adults with SCD presenting in SSC, correlated to hemoglobin (Hb) values, and compare them to those of the pediatric sickle cell population. The chest radiographs of 66 consecutive adults presenting to our emergency department complaining of symptoms consistent with acute SCC were retrospectively reviewed over a 12-month period. The radiographic findings were correlated with admission Hb values and compared with those of 50 children with known SCD presenting with SCC. Chi square analysis revealed no significant difference between the cardiovascular and bony findings in the adults and in those of the pediatric controls (p > 0.08-p > 1.0). However, one important difference in the two cohorts was that upper lobe infiltrates occurred exclusively in the pediatric group (p = 0.06). There was a statistically significant (p < 0.05) difference in cardiovascular and skeletal abnormalities between adults with Hb above and below the mean (8.2 g/dL). The radiographic features of adults presenting in acute SCCs are similar to those of children. Although the chest radiograph is often normal, in decreasing frequency, cardiovascular abnormalities, pneumonia sparing the upper lobes, and aseptic osteonecrosis of the shoulders and spine are not uncommon. There is a significant relationship, however, between cardiovascular abnormalities and Hb levels. PMID- 12653384 TI - Adolescents in the age of AIDS: myths, misconceptions, and misunderstandings regarding sexually transmitted diseases. AB - The purpose of this study is twofold: to evaluate the extent of knowledge possessed by young people residing in an urban sexually transmitted disease (STD) and AIDS epicenter about STDs, including AIDS; and to determine whether knowledge levels varied by age, gender, race/ ethnicity, and/or previous health instruction. A total of 867 adolescents (472 females and 395 males) attending a large public high school in New York City completed a self-administered survey. Levels of knowledge about AIDS transmission and prevention were high (mean percentage correct = 91.8%). Nonetheless, adolescent respondents locked awareness about the prevalence of common STDs, had limited understanding of the ways in which these diseases can be transmitted and prevented, and were unaware of potentially serious sequelae resulting from exposure to infectious agents (e.g., infertility from chlamydial infections). Young people who had taken a health education course in which STDs were discussed did slightly better on the knowledge survey than did their peers. While the prevention of HIV infection is, and should be, a national priority, more concerted efforts are needed to better educate young people about other STDs in the overall context of sexual health. PMID- 12653386 TI - Models to increase enrollment of minority females in science-based careers. AB - Enrollment of African-American females in academic pathways that lead to science based-careers may be limited by gender discrepancies in standardized test scores, academic preparation, selecting "college-prep" course work in the high school curriculum, and lack of mentors or role models. Barriers related to teacher characteristics are: (a) lack of self-confidence, (b) learning environment, (c) teacher behavior, (d) lack of female role models, and (e) failure to see the relevance between the classes and a female's expected role in life. Other limiting factors include differences in access to educational resources, differences in economic status, differences in interest or choice, cultural barriers, and lack of encouragement. Confidence building models that improve enrollment of minority females in science-based careers include equitable teaching instruction, inquiry-based pedagogy, and cooperative learning. Practices that correlate with achievement and success include challenging curricula, a nurturing learning environment, high expectations, community service, research experiences, and mentoring relationships. PMID- 12653385 TI - Benign subarachnoid space enlargement of infancy. AB - Subarachnoid space enlargement is a benign clinical entity characterized by rapid head enlargement in an infant with normal neurodevelopment. We report on two infants who had rapid increases in head circumference, family histories of macrocephaly, and normal neurodevelopment. Radiologic investigations in both infants showed subarachnoid space fluid collection but normal ventricular size. They both had a benign clinical course with resolution of the subarachnoid space fluid collection by the second year of life. The head circumference, however, remained at or above the 95th percentile. There is a need for pediatricians to be aware of this clinical entity and its benign nature. PMID- 12653387 TI - Asthma in inner cities. AB - While the management of asthma has improved over the past two decades, the incidence of asthma in the inner city has not. The inner city, comprising a disproportionate number of people who live close to or below the poverty line, shows increased rates of morbidity and mortality from asthma. African Americans and Hispanic Americans are two to six times more likely to die from asthma than their white counterparts. When federally funded programs have targeted reducing morbidity and mortality in children from these populations, they have succeeded, but in a national study only 18 states had initiatives targeting asthma in low income populations. This is tantamount to a public health crisis. Patients are not always properly diagnosed and are often without a regular source of health care, and symptoms are seen only in an acute context. Living conditions for the inner-city child have significant allergen triggers associated with house dust, cockroaches, cigarette smoke, chemical pollutants, and particulate matter. Viral infections, such as those caused by respiratory syncytial virus, are worse in crowded living conditions. The desirability of an increased public awareness of the seriousness of the disease and the need for chronic health care are issues that should be raised, through culturally relevant public means and in the knowledge that visual information is most effective. Physicians must understand the proper use of rescue and controller drugs, and asthma education must expand beyond doctors and nurses in their offices. The National Medical Association is committed to doing this aggressively, and community organizations, alliances, and coalitions must also aggressively follow. Public agencies must be lobbied to set high standards for proper asthma care and resources. With organizations acting in concert, the mortality and morbidity from asthma can be substantially prevented in the inner city. PMID- 12653388 TI - A new look at the pathophysiology of asthma. AB - Episodes of asthma, characterized by bronchial hyper-responsiveness and airway obstruction, are usually reversible and can be mitigated or prevented by appropriate use of therapeutic agents to manage asthma attacks. This article will highlight the various forms of asthma, either chronic or induced, and review both the correlating environmental factors and the immune mechanisms that are now understood to drive the inflammatory processes in the airways of the affected individuals. A significant fraction of humans is genetically predisposed toward having a robust response to allergens, characterized by a strong immunoglobulin E response (termed atopy) that manifests as an asthmatic reaction to airborne allergens in these sensitized individuals. However, occupation chemicals, airborne particles, and common insect allergens at home can also elicit an asthmatic response in exposed individuals, regardless of whether they are genetically susceptible or not. The complex immune mechanisms and sequelae that are associated with, and probably causal to, acute and persistent asthma episodes indicate that an imbalanced immune system is the primary driving force underlying asthma. This article will discuss how a dysregulated immune system, if not controlled by proper and aggressive therapeutic regimens, leads to pathogenic infiltration of the lung by various pro-inflammatory cells and eventual remodeling of the airway tissues, compromising normal lung function and one's quality of life. PMID- 12653389 TI - Best practices for pediatric asthma: improved clinical management for the inner city patient. AB - Asthma patients who live in urban areas face many challenges that contribute to and influence poor outcomes. Many inner-city asthma patients are burdened with significant barriers to care, which include difficulty in obtaining short- and long-term treatment and misperceptions about the chronic nature of the disease. Poor access to care, poor quality of care, or both have been shown to contribute to the high morbidity rate in this population. The objective of asthma therapy is to interrupt allergen/antigen exposures wherever possible and to control airway inflammation. The goal of an effective asthma program is to prevent exacerbations of the disease, achieve and maintain normal activity levels and normal or near normal lung function, and achieve a personal best for each patient with an effective action plan. An effective action plan is often difficult to implement because it takes much effort from both physician and patient to incorporate complete treatment strategies. Nevertheless, over time these plans can be very effective. The 1997 NIH report provides specific guidelines to help accomplish these goals. PMID- 12653390 TI - New treatments for asthma: the role of leukotriene modifier agents. AB - The three leukotriene (LT) modifiers approved for use in the United States, zileuton, zafirlukast, and montelukast, are the first new class of therapeutic agents to be introduced in 20 years for the treatment of asthma. These agents are referred to as leukotriene modifiers and have clearly demonstrated the ability to ameliorate bronchoconstriction and indices of airway edema and abnormal mucus production as observed in clinical trials. These agents have been shown to improve airflow, reduce the need for an inhaled bronchodilator, and improve nocturnal awakenings and asthma symptom scores appreciably. When combined with inhaled corticosteroid therapy, they may either provide additional improvement in efficacy or permit the reduction of the dose of inhaled corticosteroid necessary for effective therapy. Physicians should be familiar with these agents and consider them for use in their practice, in conjunction with current asthma management guidelines. PMID- 12653391 TI - Pediatric asthma self-management: current concepts. AB - The concept of asthma self-management began in asthma camps in the 1970s. Today all asthma camps are required to provide an educational asthma self-management program. The interaction between children and educators is brief, and if the children do not continue in an associated program after camp, the benefits may be lost. Open Airways, the first program developed specifically for minority children, has been the prototype for community asthma self-management. School based intervention programs have incorporated asthma education into the health curriculum. Some asthma education programs include an emphasis on the environment. Another approach is to develop intervention projects with parents, as in the Head Start program. This program has been very effective in increasing early recognition of asthma and decreasing recidivism in a high-risk population. Another type of project addressed the reading ability and reading comprehension of asthmatic children. Improvement in reading skills resulted in a 47% decrease in asthma recidivism. After 18 months, there were only two hospitalizations among the enrolled participants. Asthma self-management programs that are most effective for inner-city children provide an interactive, culturally relevant form of asthma education and address issues such as literacy and continuity. PMID- 12653392 TI - Racism in medicine: planning for the future. AB - African Americans and other minority populations have not benefited to the same degree as the majority of Americans from the advances in health care in our country. The vestiges of racism have resulted in inequities which exist in training, medical practice, medical decision making, the work environment and biomedical research. Recommendations are made to eliminate racial and ethnic bias in healthcare through monitoring and increasing the number and the distribution of medical trainees proportionate to the diversity of the U.S. population; through improvements in the medical school curriculum to include the impact of race and ethnicity on healthcare outcomes as well as the effect of physicians' attitudes on health; through the inclusion of an adequate number, mix and distribution of health care professionals relative to the diversity of the participants in health plans with due process for patients and physicians; and through increased participation in peer review research funding, training and as subjects in biomedical research directed to improve the quality of healthcare for diverse populations. PMID- 12653393 TI - Impact of biomedical research on African Americans. AB - Pharmaceutical development and medical research continues at a fevered pitch. Historically, however, African Americans and other minorities have not been adequately represented in the studies determining a drug's safety and efficacy in humans. A history of misuse in the medical research systems (most notably the Tuskeegee study of syphillis in a population of illiterate, poor black men) have left many blacks wary of the health care system. However, attempts to address the health disparities faced by African Americans must include processes for including wider representation of blacks--as patients as well as investigators- in clinical trials. PMID- 12653394 TI - Impact of anti-affirmative action on medical school enrollment. AB - The nation's medical, dental and health profession school admissions of African American and other under-represented minority students needs reassessment in view of recent challenges to anti-affirmative action policies. Data suggest that low income and medically underserved communities are more likely to be cared for by minority physicians. Experts project that the U.S. will need about twice as many African-American physicians as it now has to serve future patient needs. Currently, African Americans comprise 3% of the physician workforce. Decisive actions and policies--such as the recommendations made by the National Medical Association--are needed to ensure parity and cultural diversity in the medical workforce. PMID- 12653395 TI - Race, medicine, and health care in the United States: a historical survey. AB - Racism in medicine, a problem with roots over 2,500 years old, is a historical continuum that continuously affects African-American health and the way they receive healthcare. Racism is, at least in part, responsible for the fact African Americans, since arriving as slaves, have had the worst health care, the worst health status, and the worst health outcome of any racial or ethnic group in the U.S. Many famous doctors, philosophers, and scientists of each historical era were involved in creating and perpetuating racial inferiority mythology and stereotypes. Such theories were routinely taught in U.S. medical schools in the 18th, 19th, and first half of the 20th centuries. The conceptualization of race moved from the biological to the sociological sphere with the march of science. The atmosphere created by racial inferiority theories and stereotypes, 246 years of black chattel slavery, along with biased educational processes, almost inevitably led to medical and scientific abuse, unethical experimentation, and overutilization of African-Americans as subjects for teaching and training purposes. PMID- 12653396 TI - Race: a major health status and outcome variable 1980-1999. AB - Based on the latest available data, African Americans are faced with persistent, or worsening, wide and deep, race-based health disparities compared to the white or general population as we enter the new millennium. These disparities are a 382 year continuum. There have been two periods of health reform specifically addressing the correction of race-based health disparities. The first period (1865-1872) was linked to Freedmen's Bureau legislation and the second (1965 1975) was a part of the Black Civil Rights Movement. Both had dramatic and positive effects on black health status and outcome, but were discontinued too soon to correct the "slave health deficit." Although African-American health status and outcome is slowly improving, black health has generally stagnated or deteriorated compared to whites since 1980. There is a compelling need for a third period of health reform accompanied by a cultural competence movement to address and correct persistent, often worsening, race-based health disparities. PMID- 12653397 TI - Abnormal endothelial function in young African-American females: discordance with blood flow. AB - In this pilot study, we sought to compare the vasodilatory and hemodynamic properties of the peripheral vasculature in the forearms of young, healthy African-American females to similarly matched white females. We used high resolution ultrasound of the brachial artery to evaluate 11 African-American females and 8 white females. When normalized to nitrate-induced dilation, endothelium-dependent dilation was reduced in young African American females compared to white females (0.6 in African American females compared to 1.0 in white females). These results indicate the need for a larger study to examine this phenomenon. PMID- 12653398 TI - African-American heredity prostate cancer study: a model for genetic research. AB - A genome-wide scan of high-risk prostate cancer families in North America has demonstrated linkage of a particular marker to Chromosome 1q (HPC1). An even greater proportion of African-American families have shown linkage to HPC1. Therefore, investigators at the National Human Genome Research Institute (NHGRI) in collaboration with Howard University and a predominantly African-American group of urologists established the African-American Hereditary Prostate Cancer (AAHPC) Study Network to confirm the suggested linkage of HPC in African Americans with a gene on Chromosome 1. Blood samples from recruited families were sent to Howard University for extraction of DNA. The DNA was sent to NHGRI at NIH where the genotyping and genetic sequence analysis was conducted. Genotype data are merged with pedigree information so that statistical analysis can be performed to establish potential linkage. From March 1, 1998, to June 1, 1999, a total of 40 African-American families have been recruited who met the study criteria. Preliminary results suggest that racial/ethnicity grouping may affect the incidence and extent of linkage of prostate cancer to specific loci. The importance of these findings lays in the future treatment of genetic-based diseases. PMID- 12653399 TI - Fish consumption and hypertension incidence in African Americans and whites: the NHANES I Epidemiologic Follow-up Study. AB - We sought to test the hypothesis that increased consumption of fish is associated with decreased incidence of essential hypertension. Data on fish consumption and incidence of hypertension from a national cohort of 5,394 blacks and whites normotensive at baseline and followed 10 years in the NHANES I Epidemiologic Follow-up Study (NHEFS) were analyzed. Our results showed that whites aged 25-74 years had no significant association of fish consumption with incidence of hypertension. In black women, after adjusting for multiple risk factors, those who increased their fish intake from <1 time/week to > or = 1 time/week had RR = 0.42, 95% CI 0.22-0.81, p = 0.009. However, those with high intake both times had adjusted RR = 0.75, 95% CI 0.45-1.26, p = 0.28. No consistent significant associations of fish consumption with hypertension incidence were found, perhaps because fish consumption in this population was low. Further studies are needed in blacks. PMID- 12653400 TI - Early initiation of sex, drug-related risk behaviors, and sensation-seeking among urban, low-income African-American adolescents. AB - The purpose of this study was to examine the relationship of early initiation of sex, drug-use, drug-trafficking, and sensation-seeking among urban, African American adolescents. A longitudinal follow-up of 383 youth ages 9 to 15 years at baseline over four years with serial risk-assessments was used. Sexual experience and several drug-related risk behaviors increased significantly during the four year study interval. Sensation-seeking scores were higher after the baseline assessment among youth reporting tobacco, alcohol, and marijuana use and were higher, both at baseline and through several follow-up assessments, among youth reporting drug-selling and sexual activity. At baseline, the correlations among drug-related risk behaviors were all strong, except those between initiation of sex and drug-related risk behaviors. However, over time, early initiators of sex were significantly more likely to report involvement in substance use and drug delivery/sales than were late initiators. Youth reporting repeated involvement in drug-related activities were more likely to report intensive sexual involvement than they were to report experimental sex or no sex. Sensation-seeking scores were lower among youth reporting no involvement in risk behaviors. However, scores did not differ between youth exhibiting experimental behavior compared to youth demonstrating repeated risk involvement. These results support the need for alternative experiences for youth exhibiting high levels of sensation-seeking and the need for early drug/sexual risk prevention programs. PMID- 12653402 TI - Fallopian tube necrosis after postpartum sterilization. AB - Because it usually presents with nonspecific symptoms and occurs rarely, the diagnosis of fallopian tube torsion and necrosis is usually done at laparotomy. A 32 year-old woman returned to the hospital with right lower quadrant pain three days after a postpartum tubal sterilization procedure. Clinical, laboratory and imaging findings did not assist with the diagnosis. At laparotomy, after dissection of adhesions, a necrotic right fallopian tube was found. A salpingectomy was performed and the patient had an uneventful postoperative course. Fallopian tube torsion should be included in the differential diagnosis of pelvic pain in women. This patient has a good prognosis. PMID- 12653403 TI - HIV resistance and tolerability issues in antiretroviral therapy. AB - The therapeutic regimens used to treat patients with HIV disease involve a considerable pill burden. Patient adherence to dosing schedules and other factors have been implicated in the therapeutic failure of antiretroviral agents. Alternative medical approaches to HIV disease are problematic for clinicians and scientists because many of these herbal compounds have not been assessed in well designed research trials. A thorough investigation of alternative therapies by the medical and pharmacological sciences are needed for evaluation against existing therapies. PMID- 12653401 TI - Does breastfeeding protect children from asthma? Analysis of NHANES III survey data. AB - We sought to determine whether breastfeeding (yes/no) or its duration protects against the development of childhood asthma, its severity or age of onset. We conducted a secondary analysis of youth files of the National Health and Nutrition Examination Survey III (1988-94), and reviewed data from 6,783 children age 2 months to 6 years (3,316 breastfed), excluding children with a history of low birth weight or treatment in a neonatal intensive care unit. Study participants were breastfed an average of 157 days. The average age at onset of asthma was 14.3 months. In the logistic regression model, "ever breast-fed" was not a significant protective factor for developing asthma. Significant predictive factors were the mother's age at child's birth (beta = -0.08, p < 0.01), and a parent having asthma or hayfever (beta = 0.46, p < 0.01). In the linear regression model, the duration of breastfeeding was not a predictor for age at onset of asthma (beta = 0.01, p = 0.53). Only maternal smoking during pregnancy was a significant predictor of age at onset of asthma (beta = -7.59, p < 0.01). Breastfeeding does not appear to prevent asthma, delay its onset, or reduce its severity. However, breastfeeding is still recommended for its many other benefits. PMID- 12653404 TI - Light directed gene transfer by photochemical internalisation. AB - Numerous gene therapy vectors, both viral and non-viral, are taken into the cell by endocytosis, and for efficient gene delivery the therapeutic genes carried by such vectors have to escape from endocytic vesicles so that the genes can further be translocated to the nucleus. Since endosomal escape is often an inefficient process, release of the transgene from endosomes represents one of the most important barriers for gene transfer by many such vectors. To improve endosomal escape we have developed a new technology, named photochemical internalisation (PCI). In this technology photochemical reactions are initiated by photosensitising compounds localised in endocytic vesicles, inducing rupture of these vesicles upon light exposure. The technology constitutes an efficient light inducible gene transfer method in vitro, where light-induced increases in transfection or viral transduction of more than 100 and 30 times can be observed, respectively. The method can potentially be developed into a site-specific method for gene delivery in vivo. This article will review the background for the PCI technology, and several aspects of PCI induced gene delivery with synthetic and viral vectors will be discussed. Among these are: (i) The efficiency of the technology with different gene therapy vectors; (ii) use of PCI with targeted vectors; (iii) the timing of DNA delivery relative to the photochemical treatment. The prospects of using the technology for site-specific gene delivery in vivo will be thoroughly discussed, with special emphasis on the possibilities for clinical use. In this context our in vivo experience with the PCI technology as well as the clinical experience with photodynamic therapy will be treated, as this is highly relevant for the clinical use of PCI-mediated gene delivery. The use of photochemical treatments as a tool for understanding the more general mechanisms of transfection will also be discussed. PMID- 12653405 TI - Intraperitoneal oncolytic and tumor vaccination therapy with replication competent recombinant virus: the herpes paradigm. AB - The biological therapy of tumors using live viruses was first proposed a century ago but was abandoned due to potential virulence of wild-type strains. Thanks to advances in recombinant technology, replication-restricted strains have been genetically engineered, which replicate selectively within tumor cells. Examples include replication-competent mutants of herpes simplex virus (HSV), adenovirus, vesicular stomatitis virus, reovirus and measles virus. Replication-restricted oncolytic viruses are able to propagate selectively within solid tumor nodules exerting direct antitumor activity by killing infected tumor cells at the completion of a replicative cycle. In the process, they generate an intratumoral inflammatory response, which under the appropriate circumstances, may trigger the activation of an adaptive antitumor immune response, a process that has been named in situ tumor vaccination. Recombinant HSV may offer distinct advantages in oncolytic therapy of epithelial tumors. HSV is highly infectious to tumors of epithelial origin, resulting in high efficacy, there is considerable redundancy in HSV receptors, which makes the loss of HSV receptors by tumors due to mutations less likely and potent anti-herpetic drugs are commercially available, which may be used clinically to control undesired side effects. Herewith we describe the use of oncolytic viral therapy against intraperitoneal malignancies with special emphasis on oncolytic herpes simplex virus. We review the preclinical evidence on the efficacy and safety of intraperitoneal applications of HSV and discuss the rationale for its use for oncolytic therapy and in situ tumor vaccination of intraperitoneal tumors. PMID- 12653406 TI - Gene therapy targeting in the central nervous system. AB - Targeted gene therapy aims at achieving the expression of therapeutic transgenes in specific and restricted cell populations, thus sparing all other cells of the unwanted effects of the gene product. This strategy is particularly appealing for therapy of the central nervous system (CNS), where many different cell types exist, and where the inappropriate expression of a molecule can produce grave consequences. To accomplish the objectives of targeted gene therapy, two different approaches have been developed. The first one consists in creating vectors that will deliver the transgene exclusively to the selected cells, that is manipulating the transductional capacities of the vector, and the second one is based on the transcriptional properties of the transgene, so that it will only be expressed in cells where the appropriate transcriptional machinery is present. Reaching the goals of targeted gene expression will greatly increase the specificity and safety of gene therapy, thus getting us closer to the fulfillment of the expectations generated by this new branch of molecular medicine. PMID- 12653407 TI - Gene therapy using an adenovirus vector for apoptosis-related genes is a highly effective therapeutic modality for killing glioma cells. AB - Preclinical studies in animal models and human clinical trials have evaluated the safety and efficacy of adenoviral vectors for cancer gene therapy. These studies have indicated that gene delivery via adenoviral vectors, including p53 gene therapy, represents a promising therapeutic modality for many types of human cancers. This review focuses on novel strategies to induce apoptosis in glioma cells by transduction with adenoviral vectors carrying a variety of apoptosis related genes, including Fas ligand, Fas, FADD, caspase-8, p53, p33ING1, p73alpha, Bax, Apaf-1, caspase-9, IkappaBdN, caspase-3, Bcl-2, and Bcl-X(L). We conclude that adenoviral vector-mediated delivery of apoptosis-related genes other than p53 is a potentially useful gene therapy approach toward the treatment of human brain tumors. PMID- 12653408 TI - Gene and cancer therapy--pseudorabies virus: a novel research and therapeutic tool? AB - The past decade has been marked by significant advances in the application of gene transfer into living cells of animals and humans, with the resulting catapulting of preclinical and basic scientific concepts into therapeutic trials. A variety of virus-mediated gene delivery techniques have proved to be superior to other methodologies. This article concisely reviews the current status of gene and tumor therapy, focusing on virus-based technologies, describes the molecular biology of neurotropic herpesviruses and the application of herpes simplex virus, a relative of pseudorabies virus (PRV) in gene transfer and cancer therapy protocols. Finally, it addresses the issue of whether PRV, a nonhuman pathogen, could serve as a suitable research and therapeutic tool as concerns genetic and tumor therapy. PMID- 12653409 TI - Smoking withdrawal dynamics: I. Abstinence distress in lapsers and abstainers. AB - Smoking withdrawal reports from a clinical trial (n = 893) were submitted to hierarchical linear modeling as a cross-method replication of a heterogeneity approach to withdrawal measurement and to clarify the influence of postcessation smoking on symptom reports. Five individual difference parameters tapping distinct facets of withdrawal were derived: intercepts (mean severity), linear slope (direction and rate of change), quadratic trend (curvature), volatility (scatter) and, among lapsers, a cigarette coefficent (smoking-related deflections of symptoms). All parameters were highly variable across persons. Lapsers had more aversive symptom patterns than abstainers, and symptoms tended to be higherthan otherwise predicted on lapse days. These results reinforce the conclusion that withdrawal symptoms are highly variable and argue against discarding withdrawal data from participants who lapse. PMID- 12653410 TI - Smoking withdrawal dynamics: II. Improved tests of withdrawal-relapse relations. AB - In this article, the authors assessed whether continuously scaled symptom parameters derived from growth models (T. M. Piasecki et al., 2003) are linked to smoking at long-term follow-up by using data from a large-scale clinical trial (N = 893). Results revealed that higher withdrawal intercepts, positive linear slopes, and greater volatility were all positively associated with relapse, and cigarette coefficients (indicating smoking-induced withdrawal reduction) were negatively related to relapse. In models keyed around the first lapse to smoking, those destined to lapse reported more severe withdrawal during abstinence, and withdrawal patterns discriminated groups defined according to lapse duration. The findings complement earlier heterogeneity studies in implicating the pattern of changing withdrawal symptoms over time as a factor strongly associated with smoking relapse. PMID- 12653411 TI - Psychometrically matched visual-processing tasks in schizophrenia spectrum disorders. AB - Patients with schizophrenia show deficits in visual perception and working memory, but the relationship between these deficits has not been characterized with psychometrically matched tasks. The authors administered 2 visual discrimination and 6 recognition tasks to 43 schizophrenia spectrum patients and 22 nonpsychiatric subjects. When performing difficulty-matched tasks, spectrum subjects showed more severe impairments for motion compared with form processing. When tasks were matched on true score variance, spectrum subjects exhibited worse performance on both form and motion discrimination, and a differential deficit in motion recognition with a short display duration and long interstimulus interval. These results provide evidence of differential deficits in visual processing in schizophrenia that appear to be dependent on the temporal characteristics of the tasks. PMID- 12653412 TI - Neuropsychology, genetic liability, and psychotic symptoms in those at high risk of schizophrenia. AB - Neuropsychological assessments were compared among individuals at enhanced genetic risk of schizophrenia (n = 157) and controls (n = 34). The relationship between cognitive impairments and the presence of psychotic symptoms and measures of genetic risk was explored in the high-risk subjects. Neuropsychological differences were identified in many areas of function and were not accounted for by the presence of psychotic symptoms. Genetic liability was not associated with neuropsychological performance or with psychotic symptoms, but exploratory analysis showed some tests were associated with both liability measures. These results suggest that what is inherited is not the disorder itself but a state of vulnerability manifested by neuropsychological impairment, occurring in many more individuals than are predicted to develop the disorder. PMID- 12653413 TI - The construct validity of depressive personality disorder. AB - This study examined the construct validity of depressive personality disorder (DPD: American Psychiatric Association, 1994). Adult psychiatric outpatients (N = 900) underwent comprehensive Axis I and II evaluations and provided data on 4,768 of their 1st-degree relatives. Despite modest overlap, DPD was not redundant with any Axis I or II disorder. Participants with DPD exhibited more Axis I and Axis II comorbidity, and greater psychosocial dysfunction, than participants without DPD. Relatives of participants with DPD had higher rates of mood disorders, alcohol abuse, and antisocial personality. Results are consistent with findings of several other similar investigations. The authors argue that DPD is a valid construct and should be conceptualized as a personality disorder as opposed to a mood disorder. PMID- 12653414 TI - Working memory for visual features and conjunctions in schizophrenia. AB - The visual working memory (WM) storage capacity of patients with schizophrenia was investigated using a change detection paradigm. Participants were presented with 2, 3, 4, or 6 colored bars with testing of both single feature (color, orientation) and feature conjunction conditions. Patients performed significantly worse than controls at all set sizes but demonstrated normal feature binding. Unlike controls, patient WM capacity declined at set size 6 relative to set size 4. Impairments with subcapacity arrays suggest a deficit in task set maintenance: Greater impairment for supercapacity set sizes suggests a deficit in the ability to selectively encode information for WM storage. Thus, the WM impairment in schizophrenia appears to be a consequence of attentional deficits rather than a reduction in storage capacity. PMID- 12653415 TI - Cognitive aspects of chronic depression. AB - Previous research on chronic depression has focused on its link with other mood disorders and Axis II personality disorders. However, there are few data examining whether the cognitive perspective applies to this condition. In this cross-sectional study, 42 outpatients with chronic depression were compared with 27 outpatients with nonchronic major depressive disorder and 24 never psychiatrically ill controls on cognitive variables thought to be related to vulnerability to depression (e.g., dysfunctional attitudes, attributional style, a ruminative response style, and maladaptive core beliefs). Both depressed groups were more elevated than a never-ill comparison group. However, chronically depressed individuals were generally more elevated on measures of cognitive variables than those with major depressive disorders even after controlling for mood state and personality disorder symptoms. PMID- 12653417 TI - Self-esteem in schizophrenia: relationships between self-evaluation, family attitudes, and symptomatology. AB - Participants with schizophrenia (N = 59) were assessed on self-evaluation, symptomatology, and positive and negative affect (expressed emotion) from significant others. An interview-based measure of self-evaluation was used and two independent dimensions of self-esteem were derived: negative and positive evaluation of self. As predicted, negative self-evaluation was strongly associated with positive symptoms, a more critical attitude from family members was associated with greater negative self-evaluation, and analyses supported a model whereby the impact of criticism on patients' positive symptoms was mediated by its association with negative self-evaluation. The interview-based method of self-esteem assessment was found to be superior to the questionnaire because its predictive effects remained after depressed mood was accounted for. PMID- 12653416 TI - Factorial structure of pathological personality as evaluated by peers. AB - This study explored how individuals apply features of personality disorders (PDs) to peers. Members of groups nominated peers who exhibited symptoms for each of the 10 PDs in the DSM-IV. Data were gathered in 2 samples: 1st-year college students (n = 1,440) and Air Force recruits (n = 2,075). The peer method reliably identified group members exhibiting specific PD features. Factor analyses identified a clearly interpretable structure relevant to the pathological personality constructs being assessed. The structure replicated well across samples and showed expected relationships to broader models of normal personality. However, cross-method correlations of factor scores were only moderate, suggesting that peer reports are reliably different from self-reports regarding the presence of pathological personality traits. PMID- 12653418 TI - Automatic processing in spider phobia: implicit fear associations over the course of treatment. AB - This study evaluates the cognitive model of anxiety by investigating treatment related changes in automatic associations to evaluate schematic processing. Spider-phobic participants (n = 31) and healthy controls (n = 30) completed fear based Implicit Association Tests (IATs), which are reaction-time measures that tap implicit associations without requiring conscious introspection. The specific tasks involved classifying pictures of snakes and spiders along with semantic categorizations (good vs.bad, afraid vs. unafraid, danger vs. safety, and disgusting vs. appealing). Phobic individuals were assessed before and after group-based exposure treatment and 2 months later, controls were assessed at matched time points. Results supported clinical applications for implicit fear associations, including prediction of phobic avoidance, and treatment sensitivity of the fear- and disgust-specific automatic associations. PMID- 12653420 TI - Developmental trajectories of disruptive behavior problems among sons of alcoholics: effects of parent psychopathology, family conflict, and child undercontrol. AB - This study examined trajectories of disruptive behavior problems from preschool to early adolescence in 302 boys from a community-recruited sample of high-risk families. Growth modeling showed that paternal alcoholism was associated with elevated levels of sons' disruptive behavior problems. Family conflict predicted more disruptive behaviors at school entry and a slower rate of decline in such problems. Parent antisocial personality disorder (ASPD) exacerbated the effects of high preschool levels of sons' undercontrol on level of disruptive behaviors at school entry; this effect became progressively stronger across time. Low levels of undercontrol protected sons of ASPD parents from experiencing heightened levels of disruptive behaviors both at school entry and increasingly as sons grew older. Implications for subsequent maladjustment are discussed. PMID- 12653419 TI - Effects of nicotine deprivation on craving response covariation in smokers. AB - Most models of craving propose that when cravings are strong, diverse responses- thought to index an underlying craving state--covary. Previous studies provided weak support for this hypothesis. The authors tested whether nicotine deprivation affects degree of covariation across multiple measures related to craving. Heavy and light smokers (N = 127) were exposed to smoking cues while either nicotine deprived or nondeprived. Measures included urge ratings, affective valence, a behavioral choice task assessing perceived reinforcement value of smoking, and smoking-related judgment tasks. Results indicated higher correlations in the nicotine-deprived than in nondeprived group. The measures principally responsible for this effect loaded onto a single common Craving factor for nicotine-deprived but not nondeprived smokers. These findings suggest that, under certain conditions, measures of craving-related processes covary. PMID- 12653421 TI - Context-processing deficits in schizophrenia: diagnostic specificity, 4-week course, and relationships to clinical symptoms. AB - Previous research on schizophrenia suggests that context-processing disturbances are one of the core cognitive deficits present in schizophia. However, it is not clear whether such deficits are specific to schizophrenia as compared with other psychotic disorders. To address this question, the authors administered a version of the AX Continuous Performance Test designed to assess context processing in a sample of healthy controls, patients with schizophrenia, and patients with other psychotic disorders. Participants were tested at index (when medication naive and experiencing their first contact with psychiatric services) and 4 weeks later, following medication treatment. At index, patients with schizophrenia and the psychotic comparison group demonstrated similar impairments in context processing. However, context-processing deficits improved in the psychotic comparison group at 4 weeks but did not improve in patients with schizophrenia. PMID- 12653422 TI - Sexual orientation and suicide attempt: a longitudinal study of the general Norwegian adolescent population. AB - Past and future suicide attempt rates among gay, lesbian, and bisexual (GLB) young people were compared with those of heterosexual young people. A sample of Norwegian students (N = 2.924; grades 7-12) was followed in 3 data collection waves. Risk factors included previous suicide attempt,depressed mood, eating problems, conduct problems, early sexual debut, number of sexual partners, pubertal timing, self-concept, alcohol and drug use, atypical gender roles, loneliness, peer relations, social support, parental attachment, parental monitoring, and suicidal behavior among family and friends. When homosexual attraction, homosexual identity, and same-sex sexual behavior were entered to predict suicide attempt, only same-sex sexual behavior was significantly predictive. The increased odds could not be attributed to GLB students' greater exposure to risk factors for suicide attempt. PMID- 12653423 TI - Subjective qualities of memories associated with the picture superiority effect in schizophrenia. AB - Patients with schizophrenia (n = 24) matched with 24 normal subjects were presented with both words and pictures. On a recognition memory task, they were asked to give remember, know, or guess responses to items that were recognized on the basis of conscious recollection, familiarity, or guessing, respectively. Compared with normal subjects, patients exhibited a lower picture superiority effect selectively related to remember responses. Unlike normal subjects, they did not exhibit any word superiority effect in relation to guess responses; this explains why the overall picture superiority effect appeared to be intact. These results emphasize the need to take into account the subjective states of awareness when analyzing memory impairments in schizophrenia. PMID- 12653424 TI - Drinking to cope and alcohol use and abuse in unipolar depression: a 10-year model. AB - This study examined drinking to cope with distress and drinking behavior in a baseline sample of 412 unipolar depressed patients assessed 4 times over a 10 year period. Baseline drinking to cope operated prospectively as a risk factor for more alcohol consumption at 1-, 4-, and 10-year follow-ups and for more drinking problems at 1- and 4-year follow-ups. Findings elucidate a key mechanism in this process by showing that drinking to cope strengthened the link between depressive symptoms and drinking behavior. Individuals who had a stronger propensity to drink to cope at baseline showed a stronger connection between depressive symptoms and both alcohol consumption and drinking problems. PMID- 12653425 TI - An experimental test of the effect of weight-loss dieting on bulimic pathology: tipping the scales in a different direction. AB - Although it is widely accepted that dieting increases the risk for bulimic pathology, this hypothesis has not been tested in a randomized experiment. Accordingly, the authors conducted an experimental test of the dietary restraint model by randomly assigning nonobese women (N = 82) to either a 6-week, low calorie diet or a waitlist control condition. The diet intervention resulted in significant weight loss, confirming that dieting was successfully manipulated. Contrary to the restraint model, dieting resulted in significant decreases in bulimic symptoms relative to the control condition. Results converge with past findings from randomized obesity prevention and treatment trials and provide evidence that dieting does not promote bulimic pathology; rather, effective decreases in caloric intake appear to reduce bulimic symptoms. PMID- 12653427 TI - Food structure--the need for consolidated research. PMID- 12653426 TI - Using a modified Stroop task to implicitly discern the cognitive organization of alcohol expectancies. AB - Alcohol expectancies have been statistically modeled as memory networks that influence future consumption. To test the organization of expectancies suggested by these models, a modified Stroop color-naming task incorporated expectancy words associated in past research with heavy and light drinking. Light and heavy drinkers ink-named expectancy targets after being cued with an alcohol beverage word or a nonalcohol beverage word. Consistent with predictions derived from statistical models, heavy drinkers displayed significant interference when arousing expectancy words had been primed by an alcohol beverage word, whereas light drinkers displayed significant interference when sedating expectancy targets had been so primed. These results reinforce the idea that mediation o falcohol use by expectancies may be implicit as well as explicit. PMID- 12653428 TI - Mycotoxin production in wheat grains by different Aspergilli in relation to different relative humidities and storage periods. AB - Four different Aspergilli (Aspergillus oryzae, A. parasiticus, A. terreus and A. versicolor) were grown on wheat grains underdifferent degrees of relative humidity 14, 50, 74, 80 and 90%. Samples of wheat grains were taken monthly for a period of six months and examined for mycotoxin production. A. oryzae was found to produce aflatoxins B1, B2, zearalenone, DON and T-2 toxins under elevated degrees of humidity and prolonged periods of storage. A. parasiticus produced aflatoxins B1, G1, NIV, DON and T-2 toxins in high concentrations during a period of not more than three months storage at 14% relative humidity; at an increased level of relative humidity of 74% ochratoxin A, zearalenone and sterigmatocystin were also produced at high levels. The isolate was drastic in toxin production. A. terrus produced toxins at 14% relative humidity (aflatoxin G2 and DON) at levels much higher than any other prevalent degrees of humidity. A. versicolor is highly sensitive to relative humidity and grain moisture content It produced aflatoxins B1, G1, NIV and DON at a relative humidity of 50% and another toxins (aflatoxin G2, ochratoxins A, B and zearalenone) at 74%. The microorganism can be considered a trichothecene producer under suitable relative humidity. PMID- 12653429 TI - Effect of menadione (vitamin K3) addition on lipid oxidation and tocopherols content in plant oils. AB - The effect of added menadione (vitamin K3) on stored corn and wheat germ oil on the dissolution and dimerization of natural tocopherols and autoxidation of triacylglycerols was investigated. Samples of corn and wheat germ oils pure and with added menadione were stored at +20 degrees C in brown and transparent glass bottles. During storage their peroxide value, changes of the content of fatty acids and dissolution of tocopherols were measured. Destruction of individual tocopherols in tested oils with menadione stored in both dark and transparent glass bottles was greater than in the oil samples without menadione. However, the degradation of individual tocopherols was different in each oil. Addition of menadione to the both oils resulted in accelerated the process of autoxidation of these oils. The corn oil stored in transparent bottles was oxidized faster than the one stored in brown bottles. The pro-oxidant activity of menadione was additionally activated by light. In contrast, in the case of germ oil, the process of autoxidation was very fast regardless of sort of container. Addition of menadione to the plant oils influenced the dissolution of natural tocopherols but did not influence the dimerization of tocopherols. As from the experiment, the addition of menadione to the oils decreased their nutritive value. PMID- 12653430 TI - A partial supplementation of pasteurized milk with vitamin C, iron and zinc. AB - After supplementation trials, vitamin C, iron and zinc levels were increased by 1789%, 59% and 30%, respectively. Partially supplemented pasteurized milk could be a new alternative with its high nutritive value, good sensory properties and low cost. PMID- 12653431 TI - Presence of two forms of methylated (-)-epigallocatechin-3-gallate in green tea. AB - Crude catechins extract from Chinese green tea were fractionated using Sephadex LH-20 column chromatography. The fraction containing (-)-epigallocatechin-3 gallate (EGCG) was then subjected to a semipreparative high-performance liquid chromatography (HPLC). Using a mobile phase of water: dimethyl formamide: methanol: acetic acid (157:49:2:1 v/v/v/v), the mixture of two methylated catechins was separated and isolated. According to mass spectrometry (MS) and nuclear magnetic resonance ('H-NMR) date, these compounds were identified as (-) epigallocatechin-3-(3-O-methylgallate) and (-)-epigallocatechin-3-(4-O methylgallate). PMID- 12653432 TI - Effect of temperature on sorption isotherms of Brussels sprout. AB - The effect of temperature (0, 20 and 40 degrees C) on the sorptivity of freeze dried Brussels sprout was studied. It was checked which of the sorption isotherm equations that included temperature was the best to estimate experimental data. The surface area of a monolayer and isosteric heat of sorption was calculated. From the tested equations the Guggenheim-Anderson-de Boer (GAB) and Oswin equations appeared to be the best for describing sorption isotherms of freeze dried Brussels sprout. The Clausius-Clapeyron equation was found suitable for the calculation of pure isosteric heat of sorption. PMID- 12653433 TI - Biochemical and physical properties of some walnut genotypes (Juglans regia L.). AB - Walnut (Juglans regia L.) is a plant that has significant economical value and medicinal importance for human health. It is consumed in large quantities by people, therefore, it has a very important place in the public nutritive habits. Turkey has a very good walnut population of walnut varieties when compared to other parts of the world. Our objective was to investigate kernel properties and fatty acid composition, proximate composition and minerals of genotypes that are commonly grown in Turkey. Investigated genotypes were Sebin-Type-I, Korcegoz, Karabodur, Tozam and Guvenli. Physical properties are attributed to quality criteria of nuts. Sebin Type-I and Guvenli showed a good quality according to the test results. Average fatty acid compositions of genotypes (%) were 16:0; 6.88, 18:0; 7.50, 18:1; 21.16, 18.2; 43.41, 18:3; 6.25: 20:0; 11.75 for Sebin-Type-I, Korcegoz, Karabodur, Tozani and Guvenli, respectively. Proximate compounds (%) were: protein, 13.77; ash, 1.81; moisture, 2.98; fat, 62.84; total carbohydrate, 18.67. Average mineral contents (mg/100 g) were: P, 316.0; K, 270.0; Ca, 85.0; Mg, 90.0; Zn, 2.01; Mn, 2.46; Cu, 1.01; Fe, 2.90; B, 1.03. PMID- 12653434 TI - Effects of ochratoxin A on micronucleus frequency in human lymphocytes. AB - Ochratoxin A (OTA), a nephrotoxic and carcinogenic mycotoxin, was investigated to examine its potency to induce micronuclei (MN) in cultured human lymphocytes. Lymphocyte cultures were treated for the last 48 h with OTA at concentrations of 25 microM, 10 microM, 1 microM, 100 nM, 10 nM, 1 nM, and 100 microM and absolute ethanol. At the highest concentration, OTA was found to induce MN in cytokinesis blocked lymphocytes (p < 0.05). The 25 microM OTA concentration also led to a clear decrease in the percentage of binucleated cells, probably due to cytotoxicity. OTA at the other concentrations tested did not induce MN frequency. These results indicate that a high concentration of OTA is genotoxic in cultured human lymphocytes. PMID- 12653435 TI - Application of regression analysis as a method to determine total ash content in some selected nectar honeys. AB - We developed a new, indirect method for the determination of mineral substances, expressed as total ash content in bee honey varieities, based on a multiple regression model. This time-saving and effective method could serve as a new procedure in routine quality control plans of bee honey varieties. PMID- 12653436 TI - Identification of honey types. AB - In order to develop a procedure to identify a type and variety of honeys, basing on the physicochemical parameters of honey quality, two-phase calculations were conducted. A method of discrimination analysis was used. Moreover, a model to use while identifying honeys was proposed. PMID- 12653437 TI - Antioxidant activity of Egyptian Eucalyptus camaldulensis var. brevirostris leaf extracts. AB - Leaves from Eucalvptus camaldulensis var. brevirostris trees, planted in the Nile delta in Egypt, were examined for the antioxidant activity of their nonvolatile compounds. The extracts obtained by ethanol digestion and by supercritical fluid extraction (SFE; CO2 with 15% ethanol) showed the most promising antioxidative activities. In order to identify the most active compounds, both extracts were subjected to a semipreparative reversed-phase HPLC separation, the main fractions were collected, tested for antioxidative activity and analysed by different chromatographical and spectroscopical methods for identification of the most relevant compounds. Gallic and ellagic acid were found to be the prevailing antioxidants in the ethanolic extract. The main two compounds of the SFE extract with antioxidative activity revealed to be flavones. To a high degree of probability they were identified as 5-hydroxy-7,4'-dimethoxy flavone and 5 hydroxy-7,4'-dimethoxy-8-methyl flavone, respectively. The extracts obtained by ethanoldigestion were dried and administered to rats for toxicity evaluation (up to 3 g/kg body weight). No mortality was observed which indicates a very low lethality of the tested extract. PMID- 12653438 TI - Effect of hydrothermal processing on carrot carotenoids changes and interactions with dietary fiber. AB - The aim of the studies was to determine the effect of different methods of heat treatment on carotenoids changes and their interactions with insoluble and soluble dietary fiber. Three industrial varieties of carrot--Simba, Caropak and Fayette constituted the experimental material. Carrot cubes were subjected to heat treatment by putting in water with or without citric acid, or in a convection-type steam furnace. The total content of alpha- and beta-carotene was determined in all kinds of pureed carrots. Its amount bounded with insoluble dietary fiber and pectins was also determined. Changes in soluble and insoluble fractions of dietary fiber during hydrothermal treatment were also determined. It was found that the content of trans alpha- and beta-carotene in carrots decreased significantly (p < 0.05) after heat treatment, compared with the control sample. The loss observed during heat treatment in water was higher (up to 50%) than in the case of a convection-type steam furnace. The highest decrease in the content of insoluble fraction of dietary fiber and the highest increase in soluble fraction were observed after treatment with the use of steam. An analysis of interactions between carotenoids and dietary fiber fractions after hydrothermal processing shows their stronger affinity to forming bonds with pectins than with insoluble fiber. It was also found that the effect of heat treatment parameters was significant--the highest (by six times) increase in the content of beta carotene bounded with pectins was noted in pureed carrots processed in a convection-type steam furnace. PMID- 12653439 TI - Effect of selected commercial substances with cryoprotective activity on the quality of mechanically recovered, washed and frozen stored poultry meat. AB - Investigations were conducted on mechanically recovered poultry meat (MRPM) and on protein preparation obtained from MRPM by washing it first with 1% water solution of sodium chloride and with water afterwards. The raw materials were frozen at the temperature of -23 C. The effect of added stabilizers on the quality of gels produced from fresh raw materials, and after freezing and frozen storage was assessed. The following additives were used: 1% pork hydrolizate (Pork Stock), 0.5% Cremodan containing carrageens, and 1.5% bovine blood plasma (AMP 600N). Freezing and frozen storage caused a significant reduction of functional properties of MRPM and its protein preparation. None of the examined additives protected simultaneously all the investigated functional properties of the frozen samples. The amount of thermal drip, the gel texture and the amount of protein transition heat were determined by scanning differential calorimetry. The lowest thermal drip in gels obtained from frozen-stored samples was observed when bovine blood plasma was used as a stabilizer. On the other hand, the most advantageous protective effect on the proteins of the frozen MRPM and on the preparation, determined by mechanical strain resistance of the gels, was found with 1% pork hydrolizate added. The results of thermodynamic investigations of proteins revealed that the best protective effect on the frozen preparation was observed with 1.5% blood plasma added. No protective activity of added Cremodan on proteins of the frozen protein preparation was noted. PMID- 12653440 TI - Assessment of the agronomic and technological characteristics of Triticum turgidum ssp. dicoccum Schrank and T. spelta L. AB - Recently, the peoples interest in natural and organic products led to a "rediscovery" of hulled wheat, particularly T. turgidum ssp. dicoccum, commonly known as "farro". Compared with durum wheat (T. turgidum ssp. durum), farro is characterized by low productivity and low quality semolina and flour, which makes it unsuitable for the production of bread and pasta. In this paper, the agronomic, technological and nutritional characteristics of three new varieties of farro obtained by crossing the T. turgidum ssp. dicoccum c.v. Molise with T. turgidum ssp. durum c.v. Simeto, were analysed. Data were then compared with those concerning older populations of T. turgidum and of two varieties of T. spelta. The aim of this work is to highlight the productive advantages of the new varieties and to emphasize their prospective use in the production of bread and pasta with considerable health properties due to nutritional characteristics of hulled cereals. PMID- 12653441 TI - Production and physicochemical characterization of resistant starch type III derived from pea starch. AB - Smooth pea starch was used for the production of physiological important resistant starch type III. For reduction of the molecular weight of the starch, different strategies including enzymatic debranching and acid hydrolysis (lintnerization), were tested to obtain an optimal starting material for retrogradation. The resulting polymer chain lengths were analyzed by high performance anion-exchange chromatography. Temperature regimes and starch concentrations in gel were optimized during the retrogradation with the aim to obtain a high yield of resistant starch. Optimal conditions led to resistant starch contents up to 74%. The products were thermostable and showed no loss of resistant structures after autoclaving. The peak temperatures of the thermal transition were at approximately 147 degrees C. The resulting resistant starch products are suitable for the generation of functional foods. PMID- 12653442 TI - Gelatinization and pasting properties of rice starch modified with 2-octen-1 ylsuccinic anhydride. AB - Rice starch was modified with various levels of 2-octen-1-ylsuccinic anhydride (OSA). Treatments with OSA at 3, 5, and 10% resulted in starch derivatives with 0.016, 0.033, and 0.070 degrees of substitution (DS), respectively. Thermovisco properties of the derivatives were investigated by differential scanning calorimetry (DSC) and rapid visco analysis (RVA). Water content in the sample was found to have a significant effect on the characteristics of the DSC endotherm. Pasting properties of the OSA-starch and the effect of pH and salt on the RVA profiles were also studied. In general, with increased OSA-modification, the starch derivatives swelled and gelatinized at lower temperatures to achieve higher viscosities. Specifically, based on DSC analysis at 80% water, the peak temperature of gelatinization decreased from 68.5 to 63.2 degrees C as the OSA modification increased in DS from 0 (intact starch) to 0.070. On the other hand, RVA results indicate that, for samples undergoing similar increase in OSA modification, the pasting temperature decreased from 88.7 to 51.5 degrees C and the peak viscosity increased from 668 to 6822 cP. PMID- 12653443 TI - Confidentiality. PMID- 12653444 TI - Barriers to higher education for individuals with psychiatric disabilities. AB - To study barriers to higher education, this qualitative study explored the college experiences of 35 people with psychiatric disabilities. Academic performance was related to psychiatric symptoms which subsequently led to college attrition. However, many research participants showed remarkable persistence in pursuit of academic goals. Campus-based support services were rarely utilized. Implications for psychiatric rehabilitation practice are discussed. PMID- 12653445 TI - Practices differentiating high-performing from low-performing supported employment programs. AB - This paper reports on a qualitative study designed to uncover the practices that differentiate programs producing high rates of competitive employment from those with low rates. The study found 13 differentiating practice factors in three areas: building consumer interest in working, getting a job, and maintaining a job. The congruence between these findings and current definitions of "best practice" is discussed. PMID- 12653446 TI - Representation of the governed: leadership building for people with behavioral health disorders who are homeless or were formerly homeless. AB - Many organizations that provide services to individuals with behavioral health disorders are required to include people with psychiatric disabilities on their boards and action groups, yet this requirement rarely results in successful, ongoing representation. We report on a pilot project that trained people who were homeless and formerly homeless, most of whom were diagnosed with behavioral health disorders, for internships on boards and action groups that provide services to people who are homeless. We relate the project's goals to the theme of empowerment, present our findings, discuss key implementation issues, and offer recommendation for future program efforts and research. PMID- 12653447 TI - Mental health program monitoring: towards simplifying a complex task. AB - Finding measures that can assess areas of expected program impact, provide valid results, and be easily integrated into routine program practices is a significant challenge. This paper is intended to assist program staff by providing an accessible inventory of measures appropriate for routine monitoring of the status and outcome of individuals using mental health outpatient and community programs. The inventory is not exhaustive, but rather includes examples of solid measures for assessing outcomes in four key domains--symptoms, functioning, quality of life, and satisfaction. These can provide a core of information, to which measurement of more in-depth issues can be added to address specific concerns. PMID- 12653448 TI - Payee relationships: institutional payees versus personal acquaintances. AB - Most patients' payees are friends or family members, but quantitative evaluations of patients' experience of payeeship published thus far have involved patients whose payees were mental health staff. We compared ratings of different aspects of payeeship among veterans whose payees were family or friends (n = 27) and among those with institutional payees (lawyers, accountants, or clinic affiliates, n = 22). After adjusting for differences in sociodemographic characteristics, the two groups had small and statistically non-significant differences on measures of their interaction with their payees. Overall satisfaction with payees and trust in them was high. PMID- 12653449 TI - A two-faceted work participation approach to employment and career development as applied to persons with a psychiatric disability. AB - The work histories of persons with severe and persistent psychiatric disabilities are generally characterized by sporadic employment interspersed with long periods of unemployment during which they are supported by and often become dependent upon such forms of public aid as SSI or SSDI. This paper first describes a multifaceted work participation approach to employment and career building. It then explores the implications that this approach has for establishing future directions for research and interventions aimed at improving the quantity and quality of the participation in the work life domain of persons with a psychiatric disability. PMID- 12653450 TI - Aging women with schizophrenia. AB - The purpose of this study was to describe the aging experiences of women with schizophrenia. The research focused on how participants viewed their own aging with schizophrenia, their perceived worries and concerns and how they were coping with aging with the disorder. Using a qualitative approach, data were collected using multiple in-depth interviews with six participants selected purposefully from the client list of a community mental health center. Interview transcriptions were coded and analyzed according to the study questions using QSR Nudist 4 software. Several categories and sub-categories emerged. These included the improvement in the illness over time; physical and daily living activity limitations; specific positive and negative changes that the women report have accompanied aging; the profound losses experienced by the participants when they were younger as a result of having schizophrenia; and how these losses have affected their present lives in terms of limiting available informal support, creating dependency on formal programs and services, and participants' fears of the future. Based on the study findings, implications for mental health practice and services are considered and suggestions are made to guide future research. PMID- 12653451 TI - Brazil's mental health adventure. AB - This is an account of my trips to Brazil in 2001 where I worked on a series of mental health projects with Brazilian colleagues. I first got interested in Brazil after I graduated from college when I was a Peace Corps volunteer in Northeast Brazil (Bahia state). After I got out of the Peace Corps I moved to Rio de Janeiro and went to work for United Press International (UPI) in their Rio bureau. I was UPI foreign news correspondent for a year and a half. Those years in Brazil were probably the happiest years of my life. Later on, after I became ill in the U.S., my Brazilian connection played an important role in my recovery. Raised in a Victorian family in a small town in the Midwest, and schooled in a traditional boarding school for boys and then at an all men's college, Brazil's lively Latino culture served as a healthy antidote for my tendency to be reserved and often depressed. My contact with Brazilians and Brazilian culture always beckoned me on. I maintained contact with my friends in Brazil and they stuck by me through my illness years. What seemed like my emotional and intellectual "excess" to me, was easily accepted by my Brazilian friends. I felt much more myself interacting with Brazilians and connected to a larger sense of self I developed in Brazil. I traveled to Brazil at every opportunity and made friends with Brazilians I met in the States. I initiated Portuguese classes at John Carroll University in Cleveland, Ohio in the early 1990s and then was invited to teach Brazilian culture to undergraduates. These appointments and my own resilience moved me past one depression and a dysthymia condition and into the wider community. I regained my confidence as a teacher, a role I had before and during the years of my illness. From this position, I organized a club for Brazilian students studying in the Cleveland area. After this teaching stint, I felt ready to pursue full time employment and began a job search that would eventually land me in New Haven at the Connecticut Mental Health Center. Since 1997, I've spent my vacations traveling and working in Brazil as an Outside Consultant on mental health projects with colleagues in Rio and Sao Paulo. In my travels I've been befriended and supported by adherents of a social movement, not unlike the U.S. Civil Rights Movement, that has struggled for many years to close Brazil's long-term psychiatric hospitals, create community-based services and expand the rights of mental patients. Now I see my Brazilian connection as part of my ongoing recovery. I see myself as having the opportunity to be a link between the mental health worlds of the U.S. and Brazil. I believe the two countries have much to offer each other when it comes to mental health. PMID- 12653452 TI - Perspectives of people with psychiatric disabilities on employment disclosure. AB - The Americans with Disabilities Act (ADA) of 1990 protects people with disabilities from employment discrimination. Under the ADA, employers must accommodate the known disabilities of a qualified employee or applicant. For persons with psychiatric disabilities, which are often invisible, the individual is required to make a conscious decision regarding disclosing their disability to an employer. The decision to disclose is very complex because the person needs to consider the possibility of confronting stigma and negative stereotypes. A qualitative study including a focus group and individual interviews was conducted to gather data from people with psychiatric disabilities/labels regarding employment disclosure. Major findings include the significant impact of disability identity (does the participant think they have a disability), and the importance of appropriate job matching as a disclosure strategy. PMID- 12653453 TI - The Psychiatric Rehabilitation Beliefs, Goals, and Practices Scale: sensitivity to change. AB - This study evaluated the Psychiatric Rehabilitation Beliefs, Goals, and Practices Scale's (PRBGP) sensitivity to change. A training curriculum in the current principles and practices of Psychiatric Rehabilitation was given to 301 practitioners in Pennsylvania. Participants completed the PRBGP Scale prior to and immediately following the two-day, twelve-hour training workshop. The training curriculum was expected to convey a substantial amount of information to these participants. The comparison between the pre-training and post-training PRBGP Scale scores of the participants found a significant and substantial difference. The PRBGP Scale reflected the expected changes in the participants' information levels. This study concluded that the PRBGP Scale was sensitive to change, and it provided a reliable and valid means of assessing the effectiveness of educational curricula in psychiatric rehabilitation. PMID- 12653454 TI - Dreams. PMID- 12653455 TI - Prevention of the complications of diabetes. AB - For patients with diabetes mellitus (DM), chronic complications can be devastating. Cardiovascular illness, the major cause of morbidity and mortality among these patients, encompasses macrovascular disease, with heart attacks, strokes, and gangrene; and microvascular disease, with retinopathy, nephropathy, and neuropathy (somatic and autonomic). Macrovascular events occur earlier in individuals with DM than in people without DM, and the underlying pathologies are often more diffuse and severe. Diabetic arteriopathy, which encompasses endothelial dysfunction, inflammation, hypercoagulability, changes in blood flow, and platelet abnormalities, contributes to the early evolution of these events. Efforts are under way to determine interventions that may have the potential to prevent or halt the complications of DM. Tight glucose and blood pressure (BP) control is known to improve the vascular status of patients with DM by varying degrees. Use of anti-inflammatory drugs and lowering low-density lipoprotein cholesterol (LDL-C) levels are also useful. An emerging understanding of the importance of small, dense LDL-C and the anti-inflammatory effects of statins has provided new algorithms for primary prevention of macrovascular disease. Antiplatelet agents have also been shown to be effective in the secondary prevention of cardiovascular events. In the ideal world every risk factor would be addressed and each person with DM would have excellent glycemic control, low to normal BP, and a low LDL level, and would be taking an angiotensin-converting enzyme (ACE) inhibitor, together with a statin, aspirin, and clopidogrel. Under these near-perfect conditions, the emerging epidemic of macrovascular disease could be contained. Microvascular disease, however, is a consequence of hyperglycemia. For every 1% reduction in glycosylated hemoglobin it is possible to achieve a 22% to 35% reduction in the microvascular complications. BP control is vital and the liberal use of ACE inhibitors and angiotensin receptor blockers to slow the progression of renal disease should drastically reduce the incidence of blindness, dialysis, and amputations. This article provides an overview of prevention of macrovascular disease such as stroke, myocardial infarction, and peripheral arterial disease and microvascular complications such as retinopathy, nephropathy, and neuropathy in patients with DM. PMID- 12653456 TI - Colorectal cancer test use among persons aged > or = 50 years--United States, 2001. AB - Colorectal cancer is the second leading cause of cancer-related death in the United States. The lifetime risk for having colorectal cancer diagnosed is 6%. Screening measures decrease the incidence and mortality of colorectal cancer by detecting early disease and removing precancerous lesions. The U.S. Preventive Services Task Force recommends routine cancer screening for U.S. adults aged > or = 50 years with one or a combination of the following screening options: annual home fecal occult blood testing (FOBT), sigmoidoscopy every 5 years, colonoscopy every 10 years, or double contrast barium enema every 5 years. To estimate rates and evaluate trends for colorectal cancer test use among U.S. adults aged > or = 50 years, CDC analyzed data from the 2001 Behavioral Risk Factor Surveillance System (BRFSS) on the use of FOBT and sigmoidoscopy/colonoscopy and compared the data for 2001 with those for 1997 and 1999. This report summarizes the results of that analysis, which indicate that despite small increases in the self-reported use of colorectal cancer tests, screening rates remain low. Efforts to increase awareness and encourage regular colorectal cancer screening should continue. PMID- 12653457 TI - Donated television airplay of colorectal cancer education public service announcements--United States, 1999-2002. AB - To help communicate the importance of colorectal cancer (CRC) screening, in 1999, the U.S. Department of Health and Human Services (DHHS) launched the "Screen for Life: National Colorectal Cancer Action Campaign" (SFL) (http://www.cdc.gov/cancer/screenforlife) as one of many strategies addressing the prevention and early detection of CRC. As a central part of this campaign, public service announcements (PSAs) were developed to take advantage of the influence and reach of television to encourage Americans aged > or = 50 years to get tested for CRC. This report summarizes an assessment of donated television airplay that SFL PSAs received during March 1999-February 2002. According to data obtained from Arbitron Inc., a research firm that monitors broadcast media in the United States, SFL PSAs were broadcast 41,624 times, amounting to approximately 4.3 million dollars in donated television airtime. As DHHS and others promote CRC screening, CDC will continue to release and track airplay of SFL PSAs and examine the collective influence that SFL and other educational efforts and strategies have on CRC screening rates in the United States. PMID- 12653458 TI - Poisoning by an illegally imported Chinese rodenticide containing tetramethylenedisulfotetramine--New York City, 2002. AB - Illegally imported foreign products can result in domestic exposures to unusual toxic chemicals, and health-care providers might not be able to provide appropriate therapy because the chemical ingredients might not be listed or recognized even after translation of the product label. This report describes the first known case in the United States of exposure to a Chinese rodenticide containing the toxin tetramethylenedisulfotetramine (TETS), a convulsant poison. The report of this investigation highlights the need to prevent such poisonings through increased public education, awareness, and enforcement of laws banning the importation of illegal toxic chemicals. PMID- 12653459 TI - Smallpox vaccine adverse events among civilians--United States, March 4-10, 2003. AB - During the civilian smallpox vaccination program, CDC, the Food and Drug Administration, and state health departments are conducting surveillance for vaccine-associated adverse events. In the first stage of the program, active surveillance is being conducted for potentially life-threatening, moderate-to severe, and other serious adverse events and for vaccinia transmission to contacts of vaccinees (Table). Nonserious events are reported through passive surveillance and are expected to be underreported. This report summarizes smallpox vaccine adverse events reported among civilians vaccinated as of March 7, 2003, and among contacts of vaccinees, received by CDC from the Vaccine Adverse Event Reporting System (VAERS) as of March 10. PMID- 12653460 TI - FDA licensure of diphtheria and tetanus toxoids and acellular pertussis adsorbed, hepatitis B (recombinant), and poliovirus vaccine combined, (PEDIARIX) for use in infants. AB - On December 13, 2002, the U.S. Food and Drug Administration (FDA) licensed a combined diphtheria and tetanus toxoids and acellular pertussis adsorbed (DTaP), hepatitis B (HepB) (recombinant) and inactivated poliovirus vaccine (IPV), DTaP HepB-IPV (PEDIARIX, SmithKline Beecham Biologicals, Rixensart, Belgium) for use in infants ages 2, 4, and 6 months. All components in the combined vaccine are recommended for routine use by the Advisory Committee on Immunization Practices (ACIP), the Committee on Infectious Diseases of the American Academy of Pediatrics, and the American Academy of Family Physicians. Combination vaccines decrease the number of vaccine injections. PMID- 12653461 TI - Distribution of sensory neurons of ventral and dorsal cervical cutaneous nerves in dorsal root ganglia of adult rat--a double-label study using DiO and DiI. AB - To examine distribution of sensory neurons of ventral and dorsal cervical cutaneous nerves in dorsal root ganglia (DRGs), DiO and DiI tracers were applied at the proximal section of nerves (transverse superficial cervical and anterior supraclavicular nerves were selected as ventral cervical cutaneous nerves; dorsal cutaneous branches of second, third and fourth cervical nerves were selected as dorsal cervical cutaneous nerves). Located distributions were observed in DRGs of C2, C3, and C4 (25/46 DRGs). Sensory neurons of the ventral cervical cutaneous nerves were distributed in dorso-lateral or dorso-medial portions; neurons of dorsal cervical cutaneous nerves were distributed in ventro-medial or ventro lateral portions of DRGs. Moreover, sensory neurons of transverse superficial cervical and anterior supraclavicular nerves were mainly distributed from the caudal half of C2 to whole part of C4 DRGs. Results show that there is a tendency for located distribution in two group sensory neurons; also, sensory neurons of ventral cervical cutaneous nerves have a segmental distribution, which has been verified in the brachial and lumbar plexus. PMID- 12653462 TI - Micromorphological characteristics of hepatic sinusoidal endothelial cells and their basal laminae in five different animal species. AB - The micromorphological characteristics of hepatic sinusoidal endothelial cells and their basal laminae in five different animal species (cow, sheep, guinea pig, pig, and dog) were studied by electron microscopy. Using transmission electron microscopy, a prominent basal lamina was seen only in cow and sheep. In pig and guinea pig, no prominent basal lamina was evident. Basal lamina-like material was occasionally found in dog. Immunohistochemically, basal lamina was found in cow, sheep, and dog. Using scanning electron microscopy, the size of endothelial fenestrae differed between species. These results may suggest that variation of endothelia and their basal laminae of different species is related to differences in nourishment. PMID- 12653463 TI - Development of astrocytes in the lamina cribrosa sclerae of the mouse optic nerve, with special reference to myelin formation. AB - In the mouse optic nerve, the optic nerve fiber layer in the retina, the optic papilla and the lamina cribrosa sclerae (LCS) just after penetrating the eyeball failed to generate myelin, whereas the optic nerve proper in the orbit was occupied by myelinated nerve fibers. The present study investigated development of the architecture of LCS, where the axons develop from unmyelinated to myelinated type, to elucidate how the initial part of axons was unmyelinated. At the LCS of the adult optic nerve, well developed astrocytes densely formed a cytoplasmic mesh-like frame through which unmyelinated fibers passed. The astrocytes here contained numerous and densely packed intermediate glial filaments and cell organelles. This framework formed by astrocytes appeared to be completed between 7 and 14 postnatal days before oligodendrocyte progenitors, migrated from the chiasm side, reached the proximal end of LCS, and began myelin formation. Thus the failure in myelin formation at the intraocular part and LCS possibly depended upon unsuccessful migration of oligodendrocytes beyond LCS constructed by specialized astrocytes, although other inhibitory factors for myelin formation, such as adhesion molecules distributed around LCS, may be unsolved. PMID- 12653464 TI - A rare multiple variation of the greater omentum. AB - In a dissection performed in our department, we observed multiple variations of the greater omentum. The unusual attachments of the greater omentum were identified. While the right upper part combined with ligamentum teres hepatis, the left upper part had connection with the anterolateral abdominal wall. In addition, the free lower margin of the greater omentum was bound to the front of the abdomen wall. These unusual attachments contained vessels and nerves. The greater omentum is used in reconstructive surgery of the chest wall. And its ligaments are important in abdominal surgery. Therefore, it is important to keep in mind the different attachments of the greater omentum, so that caution is required during intraabdominal surgery and also in appropriately interpreting the radiographs. PMID- 12653465 TI - Single coronary artery arising from the right sinus of Valsalva (case report). AB - A single coronary artery is a rare congenital anomaly of the coronary arteries where only one coronary artery arises from the aortic trunk by a single coronary ostium, supplying the entire heart. We report a case of a 70 years-old man with mitral valvular insufficiency and atherosclerotic right and left circumflex coronary arteries, in whom coronary angiography showed a single coronary artery arising from a single ostium in the right sinus of Valsalva (R-II-B subtype) and transverse trunk coursed between aorta and pulmonary artery. The clinical significance and subtype of the single coronary artery are discussed. PMID- 12653466 TI - Electronic states and transitions in C60 and C70 fullerenes. AB - A review of the most relevant aspects of fullerene electronic structure and spectroscopy is presented. Experimental data and their interpretation based on computational results are discussed both for fullerene C60 and C70, with particular attention to the properties of the isolated molecule. Concerning singlet state spectroscopy, it is shown that because of its high symmetry, only dipole-forbidden electronic states are found in the low excitation energy region of C60. Conversely, the lowering of symmetry in C70 leads to several complications in its electronic structure and spectroscopy, due to the presence of weakly allowed transitions in the low excitation energy region. A slightly less congested distribution of low lying excited states characterizes the triplet manifold of the fullerenes. It is concluded that while C60 is important in aiding understanding of the main features in electronic spectroscopy of fullerenes, such as the presence of strong absorptions in the high energy range, its spectra are deeply Influenced by its high symmetry and are very peculiar. On the other hand, C70, with its lower symmetry and more complex spectra, represents a more realistic model for the intricate details of the electronic structure and electronic spectroscopy of larger and smaller fullerenes and their derivatives, which are generally characterized by lower symmetry compared to C60. PMID- 12653467 TI - Effect of molecular structure on the selective phototoxicity of triarylmethane dyes towards tumor cells. AB - In response to transmembrane potentials which are negative on the inner side of both the plasma and mitochondrial membranes, cationic dyes displaying appropriate structural features naturally accumulate in the cytosol and inside the mitochondria. Because enhanced mitochondrial membrane potential is a prevalent tumor cell phenotype, a number of cationic dyes preferentially accrue and are retained for longer periods in the mitochondria of tumor cells as compared to normal cells. The opportunities brought about by this phenomenon in chemo- and photochemotherapy of neoplastic diseases is highlighted by the observation that the phototoxic effects associated with some of the cationic photosensitizers known to accumulate in cell mitochondria are much more pronounced in tumor cells than in normal cells. However, the structural determinants of selective phototoxicity towards tumor cells are not well understood, and the lack of a robust model to describe the relationship between molecular structure and tumor selectivity has prevented mitochondrial targeting from becoming a more dependable therapeutic strategy. In this report we describe how the lipophilic/hydrophilic character of a series of cationic triarylmethane dyes affects the selectivity with which these photosensitizers mediate the destruction of tumor cells. Our results indicated that only the more hydrophilic triarylmethanes show tumor selectivity, presumably because these are the only dyes capable of staining energized mitochondria with a high degree of specificity. The partition of the more lipophilic dyes into a variety of extra-mitochondrial subcellular compartments occurs with comparable efficiencies in tumor and in normal cells, and this less specific subcellular localization precludes tumor selectivity from taking place. PMID- 12653468 TI - In vitro photodynamic therapy of nasopharyngeal carcinoma using 5-aminolevulinic acid. AB - The purpose of this study was to investigate the potential use of 5 aminolevulinic acid (5-ALA, 5-amino-4-oxovaleric acid) induced protoporphyrin IX (PPIX) for photodynamic therapy (PDT) of nasopharyngeal carcinoma (NPC) and its related mechanisms of inducing cell death. PPIX biosynthesis at I to 8 h after incubation of a cultured NPC cell line (HNE1) with 5-ALA (10-5,000 microg ml(-1)) was determined via fluorescence analysis HNEI cells were irradiated at 4 h after incubation with 5-ALA (10-200 microg ml(-1)) by diode laser (lambda = 630 nm) at various energy levels (1-50 J cm(-2)). The survival rates at 6, 12, 24 and 48 h after PDT were determined by MTT assay. Mechanisms of PDT-induced cell death were investigated via Anncxin-V/propidium iodide staining and DNA electrophoresis After incubation with 5-ALA, a time- and dose-dependent increase of cellular PPIX fluorescence was recorded up to a threshold concentration of 1,000 microg ml(-1) 5-ALA, above which a decline of fluorescence intensities occurred. Similar values of PPIX-fluorescence were found at 100-1,000 microg ml(-1) of 5-ALA. Unlike sole incubation with 5-ALA or sole laser irradiation, the combination of both factors lead to a significant, concentration-, energy- and time-dependent increase of cell death (p < 0.01). At 100 microg ml(-1) ALA and 10 J cm 2 laser irradiation, cellular survival was <5% after 48 h. More than 80% of induced cell deaths thereby occurred via apoptosis within the first 12 h following irradiation; necrosis was accountable for less than 20%. High level induction of apoptosis by 5-ALA-PDT was confirmed by DNA electrophoresis. Our investigations show promising results of 5-ALA based PDT of nasopharyngeal carcinoma in vitro and set the basis for future studies in tumor models or humans, respectively. PMID- 12653469 TI - Solid state dye lasers: rhodamines in silica-zirconia materials. AB - Silica-zirconia materials as well as silica-zirconia ormosils prepared by the sol gel technique were doped with the laser dyes Rhodamine B and Rhodamine 6G and used as solid state dye lasers. The photostability and efficiency of the solid state laser samples were measured in a transverse pumping configuration by either a nitrogen laser or the second harmonic of a Nd-YAG laser. Under the excitation of a nitrogen laser the photostability of Rhodamine B in silica-zirconia materials was low and decreased with a growing amount of zirconia. The photophysical properties of the incorporated dyes were studied by time-resolved fluorescence spectroscopy. The fluorescence lifetimes of both dyes increased when the matrix was modified by organic compounds Furthermore, the threshold energy of Rhodamine 6G in two ormosils containing 3 and 50% methylsilica was measured. The results revealed that the threshold energy was lower for the matrix with a higher amount of ormosil while the slope efficiency was higher in the matrix containing 30% ormosil. PMID- 12653470 TI - Climate change and skin cancer. AB - Depletion of the ozone layer and climate change by the increasing greenhouse effect are distinctly different processes. It is becoming quite clear, however, that the two global environmental problems are interlinked in several ways [D. L. Albritton, P. J Aucamp, G. Megie, R. T. Watson, Scientific Assessment of Ozone Depletion, 1998, World Meteorological Organization, Global Ozone Research and Monitoring Project, Report No. 44 (WMO, Geneva, 1998)]. In the present analysis we deal with the possibility of such an interlinkage within one effect on human health, namely, skin cancer. The increase in the incidence of skin cancer is one of the most extensively studied effects of increasing ultraviolet radiation by ozone depletion (F. R. de Gruijl, Skin cancer and solar radiation, Eur. J Cancer, 1999, 35, 2003-2009). We wondered if this impact could also be influenced by increasing environmental temperatures. Here we show that it is likely that such an influence will occur. For the same reason, it is likely that the baseline incidence of skin cancer will be augmented by rising temperatures, which may become significant in magnitude. PMID- 12653471 TI - Some photophysical studies of cis- and trans-urocanic acid. AB - Urocanic acid, an important human skin chromophore, undergoes a variety of photochemical transformations when exposed to the near-UV portion of sunlight and natural daylight, the principal reaction being the transformation from the stable trans- or (E)-form of the chromophore (trans-UA) to the biologically active cis- or (Z)-form (cis-UA), which is claimed to induce immunosuppression linked to the onset of skin cancer. This study is concerned with the comparative photophysical behaviour of the two urocanic acid isomers in aqueous solution using both continuous irradiation and pulsed irradiation techniques. The UV absorption maximum for both isomers occurs in the region of 270 nm with the absorption shape varying characteristically with pH, the cis-isomer showing a lower overall molar absorptivity. Both isomers exhibit weak fluorescence (quantum yields estimated to be less than 10(-4)) with each isomer showing small differences in the way in which pH and excitation wavelength influence the fluorescence emission characteristics. Pulsed nanosecond laser irradiation at 266 nm of aqueous solutions at pH 7 shows that both isomers undergo photo-ionisation with a quantum yield of 0.02 for the hydrated electron production, a quantum yield value comparable with that for photoisomerisation at this wavelength. Laser flash studies also show that the photo-ionised species reacts efficiently with oxygen (quenching rate kQ = 1.3 x 10(9) M(-1) s(-1)), while some preliminary experiments indicate that both cis- and trans-urocanic acids react with the semiquinone radical of L-3,4-dihydroxyphenylalanine (L-DOPA) with a fast reaction rate constant of approximately 5 x 10(7) M(-1) s(-1). The photophysical characteristics of trans-UA and cis-UA reported here are discussed in the context of other recent pulsed irradiation studies on urocanic acid over nanosecond and picosecond time scales, in an attempt to clarify the complex photo-behaviour of this interesting biomolecule. PMID- 12653472 TI - Studies of polyphotochromic behaviour of supermolecules by NMR spectroscopy. Part 1. A bis-spirooxazine with a (Z)-ethenic bridge between each moiety. AB - The photoproducts resulting from the irradiation of a closed colourless biphotochromic molecule formed by two spironaphthoxazine entities linked through an ethenic bridge and its model compound, a singular spironaphthoxazine joined by a double bond to a 2-methoxynaphthalene nucleus in the Z configuration have been studied by high resolution 1H NMR spectroscopy. At low temperature, a clean conversion between closed and open structures has been observed with the demonstration of mono and bi opening of the biphotochromic molecule. In contrast, studies at ambient temperature have implied an irreversible loss of the photochromic properties, with the formation of diastereoisomers, resulting from the cyclisation-oxidation reaction of the systems. But, by carrying out experiments on degassed samples, a significant reversion to closed initial structures was observed. PMID- 12653473 TI - Synthesis and properties of adducts of a hindered amine and 2 hydroxyphenylbenzotriazole as novel polymer stabilizers. AB - The synthesis of new types of stabilizers (a combination of 2,2,6,6 tetramethylpiperidine and 2-hydroxyphenylbenzotriazole in one molecule) is reported. Six polymerizable combined stabilizers as well as two unsaturated triazinyl-2,2,6,6-tetramethylpiperidines and three unsaturated triazinyl-2 hydroxyphenylbenzotriazoles as individual stabilizers were synthesized. Their copolymers and the terpolymers of the individual stabilizers with methyl methacrylate were obtained. Chemical bonding of the stabilizers in the polymer was confirmed spectrophotometrically. The influence of these additives on the photostability of the copolymers was studied. The participation of the combined stabilizers in the polymerization did not affect considerably the rate of copolymerization, the molecular weight and polydispersity of the copolymers. A significant stabilizing effect against photodegradation was determined. PMID- 12653474 TI - Oxidation of sulfides and disulfides under electron transfer or singlet oxygen photosensitization using soluble or grafted sensitizers. AB - Two different photosensitizers, 9,10-dicyanoanthracene (DCA) and benzophenone (BzO) or a silica bound derivative (BzO-Si) have been compared for the photooxidation of di-n-butyl sulfide and di-n-butyl disulfide. With either photosensitizer, sulfide photooxidation in acetonitrile leads very efficiently to sulfoxide, with sulfone and disulfides as by-products. Although an electron transfer mechanism has previously been established starting with DCA, our results are indicative of two competitive mechanisms using BzO as the photosensitizer, instead of singlet oxygen addition and electron transfer. The more sluggish photooxidation of disulfides leads to a complex mixture of products, among which n-butyl butanethiosulfonate and strong acids (alkylsulfonic and sulfuric) are the major ones. The relative ratio thiosulfonate: acids depends, among other factors, on the medium polarity with acid formation favored starting with BzO or BzO-Si in a methanol-water mixture. An electron transfer mechanism only can account for the observed products Superoxide anion, the formation of which is much easier starting from BzO than from DCA, is suggested to play a crucial role in this oxidative radical pathway. Starting from disulfides, grafted benzophenone is more efficient for acid formation than its soluble counterpart. As this photosensitizer can easily be recycled, an easy and smooth way to acid formation is thus available, provided that the reaction solvent is properly chosen. PMID- 12653475 TI - Photosensitization of DNA damage by glycated proteins. AB - Photosensitized DNA damage in skin is thought to be an important mechanism of UV phototoxicity. Here we demonstrate that proteins modified by advanced glycation endproducts (AGE-proteins) are photosensitizers of DNA damage and show that multiple mechanisms are involved in AGE-sensitization. AGE-chromophores accumulate on long-lived skin proteins such as collagen and elastin as a consequence of glycation, the spontaneous amino-carbonyl reaction of protein bound lysine and arginine residues with reactive carbonyl species. AGE-proteins accumulate in both the nucleus and the cytoplasm of mammalian cells. To test the hypothesis that protein-bound AGEs in close proximity to DNA are potent UV photosensitizers, a simple plasmid DNA cleavage assay was established. Irradiation of supercoiled phiX 174 DNA with solar simulated light in the presence of AGE-modified bovine serum albumin or AGE-modified RNAse A induced DNA single strand breaks. The sensitization potency of the glycated protein correlated with increased AGE-modification and the unmodified protein displayed no photosensitizing activity. AGE-sensitized formation of reactive oxygen species was not fully responsible for the observed DNA damage and other mechanisms such as direct electron transfer interaction between photoexcited AGE and DNA are likely to be involved. Glycated proteins in skin may equally function as potent photosensitizers of DNA damage with implications for photoaging and photocarcinogenesis. PMID- 12653477 TI - Lipid interaction differentiates the constitutive and stress-induced heat shock proteins Hsc70 and Hsp70. AB - Heat shock proteins play a major role in the process of protein folding, and they have been termed molecular chaperones. Two members of the Hsp70 family, Hsc70 and Hsp70, have a high degree of sequence homology. But they differ in their expression pattern. Hsc70 is constitutively expressed, whereas Hsp70 is stress inducible. These 2 proteins are localized in the cytosol and the nucleus. In addition, they have also been observed in close proximity to cellular membranes. We have recently reported that Hsc70 is capable of interacting with a lipid bilayer forming ion-conductance channels. In the present study, we found that both Hsc70 and Hsp70 interact with lipids and can be differentiated by their characteristic induction of liposome aggregation. These proteins promote the aggregation of phosphatidylserine liposomes in a time- and protein concentration dependent manner. Although both proteins are active in this process, the level and kinetics of aggregation are different between them. Calcium ions enhance Hsc70 and Hsp70 liposome aggregation, but the effect is more dramatic for Hsc70 than for Hsp70. Addition of adenosine triphosphate blocks liposome aggregation induced by both proteins. Adenosine diphosphate (ADP) also blocks Hsp70-mediated liposome aggregation. Micromolar concentrations of ADP enhance Hsc70-induced liposome aggregation, whereas at millimolar concentrations the nucleotide has an inhibitory effect. These results confirm those of previous studies indicating that the Hsp70 family can interact with lipids directly. It is possible that the interaction of Hsp70s with lipids may play a role in the folding of membrane proteins and the translocation of polypeptides across membranes. PMID- 12653476 TI - Chaperonin 60 unfolds its secrets of cellular communication. AB - The cell biology of the chaperonins (Cpns) has been intensively studied over the past 25 years. These ubiquitous and essential molecules assist proteins to fold into their native state and function to protect proteins from denaturation after stress. The structure of the most widely studied Cpn60, Escherichia coli GroEL, has been solved and its mechanism of protein folding action largely established. But in the last decade, evidence has accumulated to suggest that the Cpn60s have functions in addition to intracellular protein folding, particularly the ability to act as intercellular signals with a wide variety of biological effects. Cpn60 has the ability to stimulate cells to produce proinflammatory cytokines and other proteins involved in immunity and inflammation and may, therefore, provide a link between innate and adaptive immunity. Cpn60s are also thought to be pathogenic factors in a wide range of diseases and have recently been reported to be present in the circulation of normal subjects and those with heart disease. An interesting facet of these proteins is the finding that in spite of significant sequence conservation, individual Cpn60 proteins can express very different biological activities. This review discusses the work to date, which has revealed the cell-cell signaling actions of Cpn60 proteins. PMID- 12653479 TI - Regulation of heat shock proteins, Hsp70 and Hsp64, in heat-shocked Malpighian tubules of Drosophila melanogaster larvae. AB - It is known from earlier studies that the heat shock (HS) response in Malpighian tubules (MTs) of Drosophila larvae is different from that in other tissues because instead of the Hsp70 and other common heat shock proteins, Hsp64 and certain other new proteins are induced immediately after HS. In the present study, we examined the kinetics of the synthesis of Hsp70 and Hsp64 immediately after HS and during recovery from HS by 35S-methionine labeling and Western blotting. In addition, we also examined the transcriptional activity of hsp70 genes in larval MT cells at different times after HS by in situ hybridization and Northern blotting. The HS-induced synthesis of Hsp64 ceased by 1 hour of recovery from the HS when synthesis of the Hsp70 commenced. Our results revealed that the induced synthesis of Hsp64 immediately after HS was dependent on new transcription. Although the levels of Hsp70 in MT cells rapidly increased after its synthesis began during recovery, the levels of Hsp64 remained unaltered irrespective of its new synthesis occurring during or after HS. Inhibition of new Hsp64 synthesis by transcriptional or translational inhibitors also did not affect the total amount of this protein in MTs. The Hsp64 polypeptides synthesized in response to HS are degraded rapidly. Apparently, the cells in MTs maintain a balance between new synthesis of Hsp64 and its turnover so that under all conditions a more or less constant level of this protein is maintained. Although the Hsp70 synthesis started only after 1 hour of recovery, the hsp70 genes were transcriptionally activated immediately after HS and they continued to transcribe till at least 4 hours after the HS. The hsp70 transcripts in MT cells that recovered for 2 hours or longer did not contain the 3' untranslated regions (UTRs), which may allow their longer stability and translatability at normal temperature. Synthesis of Hsp70 during recovery period was dependent on continuing transcription. Assessment of the beta-galactosidase activity in 2 transgenic lines carrying the LacZ reporter gene under hsp70 promoter and different lengths of the 5'UTR suggested that the delayed translation of hsp70 transcripts in MTs is probably regulated by some elements in the 5'UTR. PMID- 12653478 TI - The heat shock proteins, Hsp70 and Hsp83, of Leishmania infantum are mitogens for mouse B cells. AB - Extending earlier studies, this report demonstrates that Leishmania infantum heat shock proteins (Hsps), Hsp70 and Hsp83, expressed as recombinant proteins fused to the Escherichia coil maltose-binding protein (MBP), are potent mitogens for murine splenocytes. The response was not due to lipopolysaccharide (LPS) because the stimulatory activity of Hsp preparations was sensitive to boiling and trypsin treatments, whereas the corresponding activity of LPS was resistant to both treatments. It was found that in vitro incubation of spleen cells with the Leishmania Hsps leads to the expansion of CD220-bearing populations, suggesting a direct effect of these proteins on B lymphocytes. In fact, splenocytes from B cell-deficient mice did not proliferate in response to the Leishmania Hsps. In contrast, spleen cells from athymic nude mice were significantly stimulated by these recombinant proteins as an indication that the MBP-Hsp70 and MBP-Hsp83 recombinant proteins behave as T cell-independent mitogens of B cells. Furthermore, both proteins were able to induce proliferation on B cell populations purified from BALB/c spleen. PMID- 12653480 TI - Toxoplasma gondii-derived heat shock protein HSP70 functions as a B cell mitogen. AB - We have investigated the role of Toxoplasma gondii-derived heat shock protein 70 (TgHSP70) as a B cell mitogen by measuring proliferative responses in vitro. TgHSP70 induced prominent proliferative responses in murine B cells derived not only from T gondii-infected but also from uninfected mice. Nude mice responded to TgHSP70; however, severe combined immunodeficiency, RAG1-/- B6, and microMT mice failed to respond. B220+ spleen cells showed marked proliferation after stimulation with TgHSP70, but neither CD4+ nor CD8+ population responded. This unresponsiveness of CD4+ and CD8- T cells to TgHSP70 was antigen presenting cells independent. These data indicate that TgHSP70 induced the proliferation of B cells but not T cells. Polymyxin B, a potent inhibitor of lipopolysaccharide (LPS), did not eliminate TgHSP70-induced proliferation. C3H/HeN mice responded well to TgHSP70 stimulation; however, C3H/HeJ mice carrying a point mutation in the Toll-like receptor (TLR) 4 failed to respond. This indicates that TLR4 is required for TgHSP70-induced B cell activation. The involvement of TLR4 in the TgHSP70-induced proliferative responses of spleen cells was also shown by the use of TLR4-/- mice. But TgHSP70-induced, but not LPS-induced, spleen cell proliferation was observed in MyD88-/- mice, indicating that the MyD88 molecule was involved in LPS-induced proliferation but not in TgHSP70-induced proliferation. PMID- 12653481 TI - Inhibition of tumor growth in mice with severe combined immunodeficiency is mediated by heat shock protein 70 (Hsp70)-peptide-activated, CD94 positive natural killer cells. AB - Previously, we reported that the major stress-inducible heat shock protein 70 (Hsp70) acts as a recognition structure for natural killer (NK) cells, if localized on the cell surface of tumor cells. Incubation of purified NK cells with low-dose interleukin (IL)-2 (100 IU/mL) plus recombinant Hsp70-protein or the immunogenic 14-mer Hsp70-peptide TKDNNLLGRFELSG450-463, termed TKD (2 microg/mL), enhances the cytolytic activity against Hsp70 membrane-positive (CX+) but not against Hsp70-negative (CX-) tumor cells. Here, we show that the cytolytic activity against Hsp70-positive tumor cells is inducible by incubation of unseparated peripheral blood mononuclear cells (PBMNC) with low-dose IL-2 plus TKD. Cell sorting experiments revealed that within the PBMNC population CD94(+)/CD3(-) NK cells, and not CD94(-)/CD3(+) T cells, mediate the cytotoxic activity against Hsp70-positive tumor cells. The antitumoral effect of PBMNC stimulated either with IL-2 plus TKD or with IL-2 alone was assessed in tumor bearing severe combined immunodeficiency/beige mice. A single intravenous (iv) injection of 40 x 10(6) IL-2 plus TKD-stimulated PBMNC (containing 5.2 x 10(6) NK cells) on day 4 results in a 60% reduction in tumor size, from 3.89 g to 1.56 g. In contrast, the adoptive transfer of the identical amount PBMNC stimulated with low-dose IL-2 only (containing 4.4 x 10(8) NK cells) reduces the tumor size only less than 10% (3.64 g). A phenotypic characterization of the excised tumors revealed that predominantly Hsp70-positive tumor cells were eliminated by TKD activated PBMNC. Kinetic studies demonstrate that the in vivo cytolytic capacity of TKD-stimulated PBMNC is dependent on the effector to target cell ratio. An iv injection of effector cells on day 1 or 2 after tumor cell inoculation results in significantly smaller tumors (0.77 g or 0.89 g) on day 21 as compared with mice that were immunoreconstituted on day 4 or 8 (1.39 g or 2.23 g). The tumor size of nonimmunoreconstituted control animals was 3.55 g. PMID- 12653482 TI - Analysis of interactions between domains of a small heat shock protein, Hsp30 of Neurospora crassa. AB - The alpha-crystallin-related, small heat shock proteins (sHsps), despite their overall variability in sequence, have discrete regions of conserved sequence that are involved in structural organization, as well as nonconserved regions that may perform similar roles in each protein. Recent X-ray diffraction analyses of an archeal and a plant sHsp have revealed both similarities and differences in how they are organized, suggesting that there is variability, particularly in the oligomeric organization of sHsps. As an adjunct to crystallographic analysis of sHsp structure, we employed the yeast 2-hybrid system to detect interactions between peptide regions of the sHsp of Neurospora crassa, Hsp30. We found that the conserved alpha-crystallin domain can be divided into N-terminal and C terminal subdomains that interact strongly with one another. This interaction likely represents the tertiary contacts of the monomer that were visualized in the crystallographic structures of MjHsp16.5 and wheat Hsp16.9. The conserved sHsp monomeric fold is apparently determined by these regions of conserved sequence. We found that the C-terminal portion of the alpha-crystallin domain also interacts with itself in 2-hybrid assays; however, this interaction requires peptide extension into the semiconserved carboxyl tail. This C-terminal association may represent a principal contact site between dimers that contributes to higher-order assembly, as seen for the crystallized sHsps. PMID- 12653483 TI - Differential heat shock gene hsp70-1 response to toxicants revealed by in vivo study of lungs in transgenic mice. AB - Members of heat shock proteins (Hsp70) family have been considered to respond to a large variety of stressful conditions. But it was suggested that, in pulmonary cells, Hsp response depends more closely on the type of stimulus. The lungs are critical organs potentially subjected to air pollution affecting respiratory function and, therefore, these organs are of particular interest with regard to the stress response. To investigate the stress dependence of Hsp70 response in lungs, we created transgenic mice where the firefly luciferase reporter gene is under the control of the murine hsp70-1 promoter and exposed them to different sublethal toxic conditions. For each condition, the level of transgene induction and pulmonary toxicity were assessed. We found that hsp70-1 promoter was stimulated by heat shock and cadmium but not by ozone, paraquat, and parathion, even if these chemicals induced respiratory distress and lung inflammation. Similar observations were made when expression of the endogenous hsp70-1 gene was analyzed, indicating that our transgenic model was accurately detecting hsp70-1 induction. Thereby, it appeared that hsp70-1 response is selective and depends on signaling pathways triggered by the toxicants rather than by their pathologic toxicity per se. Furthermore, because all the chemicals used in our study have been previously described to increase the level of oxidative stress, it indicates that there is no direct and simple correlation between hsp70-1 response and the level of oxidative stress, but more specific oxidative patterns should be involved in Hsp regulation. PMID- 12653484 TI - Association of HSP70 and genotoxic damage in lymphocytes of workers exposed to coke-oven emission. AB - Heat shock proteins (Hsps) have been reported to protect cells, tissues, and organisms against damage from a wide variety of stressful stimuli. Whether they protect against deoxyribonucleic acid (DNA) damage in individuals exposed to environmental stresses and chemical carcinogens is unknown. In the study, we investigated the association between Hsp70 levels (the most abundant mammalian Hsp) and genotoxic damage in lymphocytes of workers exposed to coke-oven emission using Western dot blot and 2 DNA damage assays, the comet assay and the micronucleus test. The data show that there is a significant increase in Hsp70 levels, DNA damage score, and micronucleus rates in lymphocytes of workers exposed to coke-oven emission as compared with the control subjects. Furthermore, there was a significant negative correlation of Hsp70 levels with DNA damage scores in the comet assay (r = -0.663, P < 0.01) and with micronucleus rates (r = -0.461, P < 0.01) in the exposed group. In the control group, there was also a light negative correlation between Hsp70 with DNA damage and micronuclei rate (r = -0.236 and r = 0.242, respectively), but it did not reach a statistically significant level (P > 0.05). Our results show that individuals who had high Hsp70 levels generally showed lower genotoxic damage than others. These results suggest a role of Hsp70 in the protection of DNA from genotoxic damage induced by coke-oven emission. PMID- 12653486 TI - Reinstatement of 2-year-olds' event memory using photographs. AB - This experiment examined 24- and 30-month-olds' understanding of photographs as reminders using a deferred imitation paradigm. The 24- and 30-month-olds visited a laboratory playroom and observed an experimenter demonstrating novel activities. Upon returning after a retention interval, half of the children viewed photographs depicting the activities they had viewed during their first visits as reminders of the event, as well as photographs of activities they had never seen. Children in both the reminder and the no-reminder groups were then asked to complete the originally modelled activities as a test of recall. Results indicate that although 30-month-olds recalled more activities than did 24-month olds, children in the reminder condition at both ages recalled more activities than children in the no-reminder control condition. Results are discussed in terms of the development of representational understanding and long-term recall in young children. PMID- 12653485 TI - Phenomenal characteristics of cryptomnesia. AB - Qualitative characteristics of cryptomnesia, or unintentional plagiarism were investigated. In Experiment 1 we compared accurate and inaccurate source attributions in terms of their level of confidence using instructions that did not require a fixed number of responses. Confidence was lower for plagiarised responses than for correct responses. Nevertheless, participants provided high ratings of certainty for a large proportion of their plagiarised responses. In Experiment 2 the phenomenological differences between plagiarised recall and veridical recall were compared by using an adaptation of the memory characteristics questionnaire (Johnson, Foley, Suengas, & Raye, 1988). Correct responses were associated with more experiential detail than plagiarised responses. However, a considerable number of plagiarised responses were accompanied by a confident memory of at least one qualitative characteristic. Results are discussed in terms of the source monitoring framework developed by Johnson, Hashtroudi, and Lindsay (1993). PMID- 12653487 TI - Representation of the inner self in autobiography: women's and men's use of internal states language in personal narratives. AB - Adult women and men differ in the affective qualities of their autobiographical reports. In the present study, we tested whether gender differences in emotional content are apparent in memories of both the remote past and the recent past, as well as whether they extend to internal states other than emotion. A total of 48 women and 30 men provided written accounts of four events from early in life (events from before age 7) and four events from later in life (events from age 7 or later). The narratives were coded for mention of emotions, cognitions, perceptions, and physiological states. Women used more emotion terms in their descriptions of events from later in life, relative to men; across life phases, similar trends were observed for cognition and perception terms, but not for physiological states terms. The category of internal states terms was found to be more coherent for women than for men. Results are consistent with suggestions that females and males experience differential socialisation regarding expression of internal states. PMID- 12653488 TI - Testing a positional model of the Hebb effect. AB - In two experiments, we investigated the hypothesis that a strengthening of position-item associations underlies the improvement seen in performance on an immediate serial recall task, for a list that is surreptitiously repeated every third trial. Having established a strong effect of repetition, performance was tested on transfer lists in which half the items held the same position as in the repeated list (S-items) and the remainder moved (D-items). In Experiment 1, S items showed a small advantage in order errors over control and D-items. A second experiment tested whether a design feature in Experiment 1 underlay this advantage. When the experimental design was better controlled, no improvement was shown for either S- or D-items over controls. These data were shown to be inconsistent with the results of computer simulations of a positional model. An alternative model is outlined. PMID- 12653489 TI - Infantile amnesia reconsidered: a cross-cultural analysis. AB - A number of theories have been offered over the past hundred years to explain the phenomenon of infantile amnesia, the common inability to remember autobiographical experiences from the first years of life. Recent comparative studies that examine autobiographical memories in different populations, particularly populations in North America and East Asia, have yielded intriguing findings that provide a unique opportunity to revisit some of the major theoretical views and to propose new accounts. In light of these findings, this article discusses five theoretical explanations for infantile amnesia, including cognitive and social discontinuity, the emergence of the self, early parent-child memory sharing, functions of autobiographical memory, and the complexity of life experience. The reconsideration of infantile amnesia from a cross-cultural perspective suggests that while the basic mechanisms and contributing factors may be universal, the specific ways in which these mechanisms and factors are manifested differ qualitatively across cultures. A theoretical approach that takes the larger cultural context into account can help us understand this long standing puzzle. PMID- 12653490 TI - Life-narrative and word-cued autobiographical memories in centenarians: comparisons with 80-year-old control, depressed, and dementia groups. AB - Centenarians provided autobiographical memories to either a request for a life narrative or a request to produce autobiographical memories to cue words. Both methods produced distributions with childhood-amnesia, reminiscence-bump, and recency components. The life-narrative method produced relatively more bump memories at the expense of recent memories. The life-narrative distributions were similar to those obtained from 80-year-old adults without clinical symptoms and from 80-year-old Alzheimer's dementia and depression patients, except that the centenarians had an additional 20-year period of relatively low recall between the bump and recency components. The centenarians produced more emotionally neutral memories than the other three groups and produced fewer and less detailed memories than the non-clinical 80-year-old sample. PMID- 12653491 TI - Remembering or knowing others? Person recognition and recollective experience. AB - Using Tulving's (1985) remember/know procedure, the present research investigated the experiential concomitants of person recognition. Noting basic differences in the manner in which the mind processes expectancy-related material, it was anticipated that facial typicality would be a critical determinant of people's recollective experiences (i.e., remembering vs knowing). In particular, it was expected that whereas remember responses would be more prevalent for distinctive than typical faces, know responses would reflect the opposite pattern. The results of two experiments provided general support for these predictions. In addition, the recollective advantage for distinctive faces was traced to the availability of attentional resources at encoding. These results are considered in the context of contemporary issues in person recognition and social cognition. PMID- 12653492 TI - The power of the spoken word: sociolinguistic cues influence the misinformation effect. AB - We investigated whether the sociolinguistic information delivered by spoken, accented postevent narratives would influence the misinformation effect. New Zealand subjects listened to misleading postevent information spoken in either a New Zealand (NZ) or North American (NA) accent. Consistent with earlier research, we found that NA accents were seen as more powerful and more socially attractive. We found that accents per se had no influence on the misinformation effect but sociolinguistic factors did: both power and social attractiveness affected subjects' susceptibility to misleading postevent suggestions. When subjects rated the speaker highly on power, social attractiveness did not matter; they were equally misled. However, when subjects rated the speaker low on power, social attractiveness did matter: subjects who rated the speaker high on social attractiveness were more misled than subjects who rated it lower. There were similar effects for confidence. These results have implications for our understanding of social influences on the misinformation effect. PMID- 12653494 TI - Sixth International Conference on Chemical Structures. PMID- 12653493 TI - British guideline on the management of asthma. PMID- 12653495 TI - Similarity searching using reduced graphs. AB - Reduced graphs provide summary representations of chemical structures. In this work, the effectiveness of reduced graphs for similarity searching is investigated. Different types of reduced graphs are introduced that aim to summarize features of structures that have the potential to form interactions with receptors while retaining the topology between the features. Similarity searches have been carried out across a variety of different activity classes. The effectiveness of the reduced graphs at retrieving compounds with the same activity as known target compounds is compared with searching using Daylight fingerprints. The reduced graphs are shown to be effective for similarity searching and to retrieve more diverse active compounds than those found using Daylight fingerprints; they thus represent a complementary similarity searching tool. PMID- 12653496 TI - Further development of reduced graphs for identifying bioactive compounds. AB - Reduced graphs provide summary representations of chemical structures. Here, a variety of different types of reduced graphs are compared in similarity searches. The reduced graphs are found to give comparable performance to Daylight fingerprints in terms of the number of active compounds retrieved. However, no one type of reduced graph is found to be consistently superior across a variety of different data sets. Consequently, a representative set of reduced graphs was chosen and used together with Daylight fingerprints in data fusion experiments. The results show improved performance in 10 out of 11 data sets compared to using Daylight fingerprints alone. Finally, the potential of using reduced graphs to build SAR models is demonstrated using recursive partitioning. An SAR model consistent with a published model is found following just two splits in the decision tree. PMID- 12653497 TI - Visualization and substructure retrieval tools in the MOGADOC database (molecular gasphase documentation). AB - Over the years the MOGADOC database, which covers the literature for gas-phase electron diffraction, microwave spectroscopy, and molecular radio astronomy from the inception of each method, has been developed. Recently visualization and substructure retrieval tools have been implemented. New features are described by means of typical database applications. PMID- 12653498 TI - Evaluation of an experimental chemistry preprint server. AB - A preprint is a research article made publicly available prior to formal publication. A preprint server is a freely available archive and distribution medium for preprints, allowing rapid dissemination and use of multimedia and supporting files. Electronic eprints have been widely adopted in certain fields (notably high energy physics), but, until recently, the preprint concept has not been received with enthusiasm by most chemists. Despite the fact that preprints have the advantage of rapid publication, chemists have been reluctant to produce them because they could be viewed as "unallowable" for research assessment or tenure exercises or for publication in certain prestigious journals. In theory, preprints, together with version control and online discussion, could be a useful compromise: rapid prepublication followed by open peer review, before publication in a traditional journal. This paper constitutes a preliminary evaluation of a Chemistry Preprint Server in its second year of operation and summarizes the lessons that can be learned from the experiment to date. PMID- 12653499 TI - Cheminformatics analysis of organic substituents: identification of the most common substituents, calculation of substituent properties, and automatic identification of drug-like bioisosteric groups. AB - A large set of more than 3 million molecules was processed to find all the organic substituents contained in the set and to identify the most common ones. During the analysis, 849 574 unique substituents were found. Extrapolated to the number of known organic molecules, this result suggests that about 3.1 million substituents are known. Based on these findings the size of virtual organic chemistry space accessible using currently known synthetic methods is estimated to be between 10(20) and 10(24) molecules. The extracted substituents were characterized by calculated electronic, hydrophobic, steric, and hydrogen bonding properties as well as by the drug-likeness index. Various possible applications of such a large database of drug-like substituents characterized by calculated properties are discussed and illustrated by reference to a Web-based tool for automatic identification of bioisosteric groups. PMID- 12653500 TI - Optimizing the size and configuration of combinatorial libraries. AB - This paper addresses a major issue in library design, namely how to efficiently optimize the library size (number of products) and configuration (number of reagents at each position) simultaneously with other properties such as diversity, cost, and drug-like physicochemical property profiles. These objectives are often in competition, for example, minimizing the number of reactants while simultaneously maximizing diversity, and thus present difficulties for traditional optimization methods such as genetic algorithms and simulated annealing. Here, a multiobjective genetic algorithm (MOGA) is used to vary library size and configuration simultaneously with other library properties. The result is a family of solutions that explores the tradeoffs in the objectives. This is achieved without the need to assign relative weights to the objectives. The user is then able to make an informed choice on an appropriate compromise solution. The method has been applied to two different virtual libraries: a two-component aminothiazole library and a four-component benzodiazepine library. PMID- 12653502 TI - Calculation of intersubstituent similarity using R-group descriptors. AB - This paper discusses the calculation of the similarities between pairs of substituents on ring systems. An R-group descriptor characterizes the distribution of some atom-based property, such as elemental type or partial atomic charge, at increasing numbers of bonds distant from the point of substitution on the parent ring. The similarity between a pair of descriptors is then calculated by a comparison of the corresponding property vectors. Experiments with the BIOSTER database demonstrate the ability of such similarity measures to discriminate between bioisosteric and nonbioisosteric functional groups. PMID- 12653501 TI - Similarity metrics for ligands reflecting the similarity of the target proteins. AB - In this study we evaluate how far the scope of similarity searching can be extended to identify not only ligands binding to the same target as the reference ligand(s) but also ligands of other homologous targets without initially known ligands. This "homology-based similarity searching" requires molecular representations reflecting the ability of a molecule to interact with target proteins. The Similog keys, which are introduced here as a new molecular representation, were designed to fulfill such requirements. They are based only on the molecular constitution and are counts of atom triplets. Each triplet is characterized by the graph distances and the types of its atoms. The atom-typing scheme classifies each atom by its function as H-bond donor or acceptor and by its electronegativity and bulkiness. In this study the Similog keys are investigated in retrospective in silico screening experiments and compared with other conformation independent molecular representations. Studied were molecules of the MDDR database for which the activity data was augmented by standardized target classification information from public protein classification databases. The MDDR molecule set was split randomly into two halves. The first half formed the candidate set. Ligands of four targets (dopamine D2 receptor, opioid delta receptor, factor Xa serine protease, and progesterone receptor) were taken from the second half to form the respective reference sets. Different similarity calculation methods are used to rank the molecules of the candidate set by their similarity to each of the four reference sets. The accumulated counts of molecules binding to the reference target and groups of targets with decreasing homology to it were examined as a function of the similarity rank for each reference set and similarity method. In summary, similarity searching based on Unity 2D-fingerprints or Similog keys are found to be equally effective in the identification of molecules binding to the same target as the reference set. However, the application of the Similog keys is more effective in comparison with the other investigated methods in the identification of ligands binding to any target belonging to the same family as the reference target. We attribute this superiority to the fact that the Similog keys provide a generalization of the chemical elements and that the keys are counted instead of merely noting their presence or absence in a binary form. The second most effective molecular representation are the occurrence counts of the public ISIS key fragments, which like the Similog method, incorporates key counting as well as a generalization of the chemical elements. The results obtained suggest that ligands for a new target can be identified by the following three-step procedure: 1. Select at least one target with known ligands which is homologous to the new target. 2. Combine the known ligands of the selected target(s) to a reference set. 3. Search candidate ligands for the new targets by their similarity to the reference set using the Similog method. This clearly enlarges the scope of similarity searching from the classical application for a single target to the identification of candidate ligands for whole target families and is expected to be of key utility for further systematic chemogenomics exploration of previously well explored target families. PMID- 12653503 TI - Searching for patterns of amino acids in 3D protein structures. AB - This paper describes the program ASSAM, which has been developed to search for patterns of amino acid side-chains in the 3D structures in the Protein Data Bank. ASSAM represents an amino acid by a vector drawn from the main chain towards the functional part of the amino acid and then computes a graph representation of a protein in which the individual side-chain vectors are the nodes and the intervector distances are the edges. The presence of a query pattern in a Protein Data Bank structure can then be searched for by means of a subgraph isomorphism algorithm. Recent enhancements to ASSAM allow searches to include the following: the main-chain structure in addition to the side-chains; the secondary structure and solvent accessibility of side-chains; allowable distances from a known binding-site; disulfide bridges; and improved generic and wild-card queries. The effectiveness of these approaches is demonstrated by extensive searches of the Protein Data Bank for typical 3D query patterns. PMID- 12653504 TI - Fast calculation of quantum chemical molecular descriptors from the electronegativity equalization method. AB - The use of the Electronegativity Equalization Method (EEM) is presented for high performance calculation of molecular electrostatic descriptors, giving quite similar results to those obtained through Density Functional Theory (B3LYP/6 31G*) calculations. Molecular descriptors include atomic charges and different related descriptors as well as Fukui functions, hardness and softness. PMID- 12653505 TI - Prediction of aqueous solubility of organic compounds based on a 3D structure representation. AB - Two quantitative models for the prediction of aqueous solubility of 1293 organic compounds were developed by a Multilinear Regression (MLR) analysis and a Back Propagation (BPG) neural network. The molecules were described by a set of 32 values of a Radial Distribution Function (RDF) code representing the 3D structure and eight additional descriptors. The 1293 compounds were divided into a training set of 797 compounds and a test set of 496 compounds based on a Kohonen self organizing neural network map. The obtained models show a good predictive power: for the test set, a correlation coefficient of 0.96 and a standard deviation of 0.59 were achieved by the back-propagation neural network approach. PMID- 12653506 TI - Combination of fingerprint-based similarity coefficients using data fusion. AB - Many different types of similarity coefficients have been described in the literature. Since different coefficients take into account different characteristics when assessing the degree of similarity between molecules, it is reasonable to combine them to further optimize the measures of similarity between molecules. This paper describes experiments in which data fusion is used to combine several binary similarity coefficients to get an overall estimate of similarity for searching databases of bioactive molecules. The results show that search performances can be improved by combining coefficients with little extra computational cost. However, there is no single combination which gives a consistently high performance for all search types. PMID- 12653507 TI - CLIP: similarity searching of 3D databases using clique detection. AB - This paper describes a program for 3D similarity searching, called CLIP (for Candidate Ligand Identification Program), that uses the Bron-Kerbosch clique detection algorithm to find those structures in a file that have large structures in common with a target structure. Structures are characterized by the geometric arrangement of pharmacophore points and the similarity between two structures calculated using modifications of the Simpson and Tanimoto association coefficients. This modification takes into account the fact that a distance tolerance is required to ensure that pairs of interatomic distances can be regarded as equivalent during the clique-construction stage of the matching algorithm. Experiments with HIV assay data demonstrate the effectiveness and the efficiency of this approach to virtual screening. PMID- 12653508 TI - Evaluation of similarity measures for searching the dictionary of natural products database. AB - Similarity searches using combinations of seven different similarity coefficients and six different representations have been carried out on the Dictionary of Natural Products database. The objective was to discover if any special methods of searching apply to this database, which is very different in nature from the many synthetic databases that have been the subject of previous studies of similarity searching. Search effectiveness was assessed by a recall analysis of the search outputs from sets of pharmacologically active target structures. The different target sets produce exceptional but contradictory results for the Russell-Rao and Forbes coefficients, which have been shown to be due to a dependence on molecular size; these are the coefficients of choice in the case of large and small structures, respectively. Rankings from these results have been combined using a data fusion scheme and some small gains in performance were normally obtained by using substructural fingerprints and molecular holograms in combination with the Squared Euclidean or Tanimoto coefficients. PMID- 12653509 TI - Efficient generation, storage, and manipulation of fully flexible pharmacophore multiplets and their use in 3-D similarity searching. AB - Pharmacophore triplets and quartets have been used by many groups in recent years, primarily as a tool for molecular diversity analysis. In most cases, slow processing speeds and the very large size of the bitsets generated have forced researchers to compromise in terms of how such multiplets were stored, manipulated, and compared, e.g., by using simple unions to represent multiplets for sets of molecules. Here we report using bitmaps in place of bitsets to reduce storage demands and to improve processing speed. Here, a bitset is taken to mean a fully enumerated string of zeros and ones, from which a compressed bitmap is obtained by replacing uniform blocks ("runs") of digits in the bitset with a pair of values identifying the content and length of the block (run-length encoding compression). High-resolution multiplets involving four features are enabled by using 64 bit executables to create and manipulate bitmaps, which "connect" to the 32 bit executables used for database access and feature identification via an extensible mark-up language (XML) data stream. The encoding system used supports simple pairs, triplets, and quartets; multiplets in which a privileged substructure is used as an anchor point; and augmented multiplets in which an additional vertex is added to represent a contingent feature such as a hydrogen bond extension point linked to a complementary feature (e.g., a donor or an acceptor atom) in a base pair or triplet. It can readily be extended to larger, more complex multiplets as well. Database searching is one particular potential application for this technology. Consensus bitmaps built up from active ligands identified in preliminary screening can be used to generate hypothesis bitmaps, a process which includes allowance for differential weighting to allow greater emphasis to be placed on bits arising from multiplets expected to be particularly discriminating. Such hypothesis bitmaps are shown to be useful queries for database searching, successfully retrieving active compounds across a range of structural classes from a corporate database. The current implementation allows multiconformer bitmaps to be obtained from pregenerated conformations or by random perturbation on-the-fly. The latter application involves random sampling of the full range of conformations not precluded by steric clashes, which limits the usefulness of classical fingerprint similarity measures. A new measure of similarity, The Stochastic Cosine, is introduced here to address this need. This new similarity measure uses the average number of bits common to independently drawn conformer sets to normalize the cosine coefficient. Its use frees the user from having to ensure strict comparability of starting conformations and having to use fixed torsional increments, thereby allowing fully flexible characterization of pharmacophoric patterns. PMID- 12653510 TI - Comparison of ranking methods for virtual screening in lead-discovery programs. AB - This paper discusses the use of several rank-based virtual screening methods for prioritizing compounds in lead-discovery programs, given a training set for which both structural and bioactivity data are available. Structures from the NCI AIDS data set and from the Syngenta corporate database were represented by two types of fragment bit-string and by sets of high-level molecular features. These representations were processed using binary kernel discrimination, similarity searching, substructural analysis, support vector machine, and trend vector analysis, with the effectiveness of the methods being judged by the extent to which active test set molecules were clustered toward the top of the resultant rankings. The binary kernel discrimination approach yielded consistently superior rankings and would appear to have considerable potential for chemical screening applications. PMID- 12653511 TI - A geodesic framework for analyzing molecular similarities. AB - A fast self-organizing algorithm for extracting the minimum number of independent variables that can fully describe a set of observations was recently described (Agrafiotis, D. K.; Xu, H. Proc. Natl. Acad. Sci.U.S.A. 2002, 99, 15869-15872). The method, called stochastic proximity embedding (SPE), attempts to generate low dimensional Euclidean maps that best preserve the similarities between a set of related objects. Unlike conventional multidimensional scaling (MDS) and nonlinear mapping (NLM), SPE preserves only local relationships and, by doing so, reveals the intrinsic dimensionality and metric structure of the data. Its success depends critically on the choice of the neighborhood radius, which should be consistent with the local curvature of the underlying manifold. Here, we describe a procedure for determining that radius by examining the tradeoff between the stress function and the number of connected components in the neighborhood graph and show that it can be used to produce meaningful maps in any embedding dimension. The power of the algorithm is illustrated in two major areas of computational drug design: conformational analysis and diversity profiling of large chemical libraries. PMID- 12653512 TI - Modeling toxicity by using supervised kohonen neural networks. AB - Counterprogation neural network is shown to be a powerful and suitable tool for the investigation of toxicity. This study mined a data set of 568 chemicals. Two hundred eighty-two objects were used as the training set and 286 as the test set. The final model developed presents high performances on the data set R(2) = 0.83 (R(2) = 0.97 on the training set, R(2) = 0.59 on the test set). This technique distinguishes itself also for the ability to give to the expert two-dimensional maps suitable for the study of the distribution/clustering of the data and the identification of outliers. PMID- 12653513 TI - The Chemistry Development Kit (CDK): an open-source Java library for Chemo- and Bioinformatics. AB - The Chemistry Development Kit (CDK) is a freely available open-source Java library for Structural Chemo- and Bioinformatics. Its architecture and capabilities as well as the development as an open-source project by a team of international collaborators from academic and industrial institutions is described. The CDK provides methods for many common tasks in molecular informatics, including 2D and 3D rendering of chemical structures, I/O routines, SMILES parsing and generation, ring searches, isomorphism checking, structure diagram generation, etc. Application scenarios as well as access information for interested users and potential contributors are given. PMID- 12653514 TI - Topological indices based on vertex, edge, ring, and distance: application to various physicochemical properties of diverse hydrocarbons. AB - This paper developed the Edge degree-Distance Index (EDI) and Sum of edges (S(e)) based on the edge and distance of molecular graph. This set of topological indices, EDI, S(e) combined with VDI, OEI, and RDI proposed in our previous paper can characterize the molecular structures of diverse hydrocarbons well. The regression analyses against nine physicochemical properties, such as boiling points (Bp), critical properties (Tc, Pc, Vc), heat capacities (Cp), and so on, of 1038 diverse hydrocarbons were investigated, and good correlations were obtained. PMID- 12653516 TI - The geometric structure of deformed nanotubes and the topological coordinates. AB - After summarizing the harmonic approach to topological coordinates and the null space embedding of graphs, three-coordinated tiling of the plane by hexagons, pentagons, and heptagons are presented and used for the construction of tubular, toroidal, and helical carbon structures. Physically realistic 3D geometries are formed from the corresponding adjacency matrices, and the final structure was obtained with the help of Brenner-potential based molecular mechanics methods. PMID- 12653515 TI - Tuning neural and fuzzy-neural networks for toxicity modeling. AB - The need for general reliable models for predicting toxicity has led to the use of artificial intelligence. We applied neural and fuzzy-neural networks with the QSAR approach. We underline how the networks have to be tuned on the data sets generally involved in modeling toxicity. This study was conducted on 562 organic compounds in order to establish models for predictive the acute toxicity in fish. PMID- 12653517 TI - Decision forest: combining the predictions of multiple independent decision tree models. AB - The techniques of combining the results of multiple classification models to produce a single prediction have been investigated for many years. In earlier applications, the multiple models to be combined were developed by altering the training set. The use of these so-called resampling techniques, however, poses the risk of reducing predictivity of the individual models to be combined and/or over fitting the noise in the data, which might result in poorer prediction of the composite model than the individual models. In this paper, we suggest a novel approach, named Decision Forest, that combines multiple Decision Tree models. Each Decision Tree model is developed using a unique set of descriptors. When models of similar predictive quality are combined using the Decision Forest method, quality compared to the individual models is consistently and significantly improved in both training and testing steps. An example will be presented for prediction of binding affinity of 232 chemicals to the estrogen receptor. PMID- 12653518 TI - On a four-dimensional representation of DNA primary sequences. AB - We consider a four-dimensional representation of DNA primary sequences by assigning to each of the four basic amino acids A, T, G, C directions along the four orthogonal coordinate axes. Advantages and limitations of the novel representation of DNA primary sequences are discussed, and the use of the 4-D representation is illustrated by constructing novel sequence invariants. Comparisons with the similarity/dissimilarity results based on 2-D and 3-D representations for a set of eight short DNA sequences corresponding to the first exon of beta globin in eight species, including human, are considered to illustrate the use of our novel sequence invariants based on the entries in derived sequence matrices restricted to a selected width of a band along the main diagonal. PMID- 12653520 TI - Quantum-mechanical QSAR/QSPR descriptors from momentum-space wave functions. AB - It is shown that quantum-mechanical descriptors obtained as parameters from the one-dimensional radial distribution function of electron momentum can be used to predict molecular activities or properties to a precision that compares favorably with the more traditional QSAR/QSPR methods. The distribution function is derived from momentum space ab initio wave functions. The predictive value of the descriptors is illustrated by their application to the estimation of McGowan's volume, gas-chromatographic retention time, gas-hexadecane partition coefficient, second hyperpolarizability, and tadpole narcotic activity. PMID- 12653519 TI - Comparison between neural networks (NN) and principal component analysis (PCA): structure activity relationships of 1,4-dihydropyridine calcium channel antagonists (nifedipine analogues). AB - The applicability of the neural network computer package PSDD (Perceptron Simulator for Drug Design/Perceptron-type Neural Network Simulator) in structure activity relationship (SAR) studies was investigated. A group of 1,4 dihydropyridine derivatives was used in order to compare the PSDD results with those obtained previously with PCA. Calculated atomic and molecular descriptors using the semiempirical AM1 method were mainly used. It was shown that the predictive capability demonstrated by PSDD in SAR analysis were almost equivalent to that of PCA. PMID- 12653521 TI - On the possibility of detecting low barrier hydrogen bonds with kinetic measurements. AB - Recent experimental evidence has pointed to the possible presence of a short, strong hydrogen bond in the enzyme-substrate transition states in some biochemical reactions. To date, most experimental measures of these short, strong hydrogen bonds have monitored their equilibrium properties. In this work we show that kinetic measurements can also be used to detect the presence of short, strong hydrogen bonds. In particular, we find nontrivial differences among rate constant ratios of protonated to deuterated hydrogen bonds between strong and weak hydrogen bonds for proton transfer between donor-acceptor sites. We quantify this kinetic isotope effect by performing dynamical calculations of these rate constants by computing reactive flux through a dividing surface. This reactive flux is computed by evolving trajectories on an effective quantum mechanical potential energy surface. PMID- 12653523 TI - A chemical class-based approach to predictive model generation. AB - We make a quantitative comparison of two distinct approaches to predictive model generation in the context of diverse screening data. In the default approach, a single recursive partitioning model is constructed using all of the training data at one time. In the "class-based" approach, the same data are first partitioned into homogeneous, scaffold-based classes, and models are constructed within each class independently. Both approaches are tested on the identical set of hold-out data, using a formal protocol that includes consensus scoring to handle the multiple class-based models. The entire process is performed using three different descriptor sets and is repeated using five separate random trials, such that the trial-averaged prediction rates for the two approaches can be quantitatively compared. We find that although the predictive performances of the class-based and default approaches are similar, the former has at least two distinct advantages. The first is greater interpretability, in that chemists can more easily extract useful structure-activity information from the models. The second is greater reliability, allowing models to be applied with increased confidence to unseen data in virtual-screening applications. PMID- 12653522 TI - Prediction of aquatic toxicity: use of optimization of correlation weights of local graph invariants. AB - Quantitative structure-activity relationships (QSARs) were developed for three sets of toxicity data. Chemicals in each set represented a number of narcoses and electrophilic mechanisms of toxic action. A series of quantitative structure toxicity models correlating toxic potency with a number of optimization of correlation weights of local graph invariants were developed. In the case of the toxicity of a heterogeneous set of benzene derivatives to Tetrahymena pyriformis, the QSARs were based on the Descriptor of Correlation Weights (DCW) using atoms and extended connectivity (EC) graph invariants. The model [log (IGC(50)(-1)) = 0.0813 DCW(a(k),(3)EC(k)) + 2.636; n = 157, r(2) = 0.883, s = 0.27, F = 1170, Pr > F = 0.0001] based on third-order EC of 89 descriptors was observed to be best for the benzene data. However, fits for these data of > 0.800 were achieved ECs with as few as 23 variables. The relationship between the toxicity predicted by this model and experimental toxicity values for the test set [obs. log(IGC(50)( 1))) = 0.991 (pred. (log(IGC(50)(-1))) - 0.012; n = 60, r(2) = 0.863, s = 0.28, F = 372, Pr > F = 0.0001] is excellent. The utility of the approach was demonstrated by the model [log (IGC(50)(-1)) = 0.1744(DCW (a(k), (2)EC) - 3.505; n = 39, r(2) = 0.900, s = 0.35, F = 333, Pr > F = 0.0001] for the toxicity data for T. pyriformis exposed to halo-substituted aliphatic compounds and the model [log (IC(50)(-1)) = 0.1699(DCW (a(k), (2)EC)) - 2.610; n = 66, r(2) = 0.901, s = 0.31, F = 583, Pr > F = 0.0001] for the Vibrio fischeri toxicity data. PMID- 12653524 TI - Assessing model fit by cross-validation. AB - When QSAR models are fitted, it is important to validate any fitted model-to check that it is plausible that its predictions will carry over to fresh data not used in the model fitting exercise. There are two standard ways of doing this using a separate hold-out test sample and the computationally much more burdensome leave-one-out cross-validation in which the entire pool of available compounds is used both to fit the model and to assess its validity. We show by theoretical argument and empiric study of a large QSAR data set that when the available sample size is small-in the dozens or scores rather than the hundreds, holding a portion of it back for testing is wasteful, and that it is much better to use cross-validation, but ensure that this is done properly. PMID- 12653525 TI - A wavelet, fourier, and PCA data analysis pipeline: application to distinguishing mixtures of liquids. AB - Using a new optical engineering technique for the "fingerprinting" of beverages and other liquids, we study and evaluate a range of features. The features are based on resolution scale, invariant frequency information, entropy, and energy. They allow mixtures of beverages to be very precisely placed in principal component plots used for the data analysis. To show this we make use of data sets resulting from optical/near-infrared and ultrasound sensors. Our liquid "fingerprinting" is a relatively open analysis framework in order to cater for different practical applications, in particular, on one hand, discrimination and best fit between fingerprints, and, on the other hand, more exploratory and open ended data mining. PMID- 12653526 TI - Version of zones and zigzag structure in icosahedral fullerenes and icosadeltahedra. AB - A circuit of faces in a polyhedron is called a zone if each face is attached to its two neighbors by opposite edges. (For odd-sized faces, each edge has a left and a right opposite partner.) Zones are called alternating if, when odd faces (if any) are encountered, left and right opposite edges are chosen alternately. Zigzag (Petrie) circuits in cubic (= trivalent) polyhedra correspond to alternating zones in their deltahedral duals. With these definitions, a full analysis of the zone and zigzag structure is made for icosahedral centrosymmetric fullerenes and their duals. The zone structure provides hypercube embeddings of these classes of polyhedra which preserve all graph distances (subject to a scale factor of 2) up to a limit that depends on the vertex count. These embeddings may have applications in nomenclature, atom/vertex numbering schemes, and in calculation of distance invariants for this subclass of highly symmetric fullerenes and their deltahedral duals. PMID- 12653527 TI - A new approach of evaluating bond dissociation energy from eigenvalue of bonding orbital-connection matrix for C-C and C-H bonds in alkane. AB - A new bonding orbital-connection matrix was constructed, in which the diagonal elements were assigned the chemical potentials E(1) and E(2) of two radicals R(i)* and R(j)* obtained by cutting the interested bond, and the off-diagonal elements, representing bonding connections, were assigned values 1. The eigenvalues X(1CC) and X(1CH) of the bonding orbital-connection matrix were obtained for C-C and C-H bonds, respectively. Also a steric effect parameter S(ij) was proposed for C-C bond. Using X(1CC), X(1CH), and S(ij) as bond descriptors, good correlations with the Bond Dissociation Energies (BDEs) of the C-C and C-H bonds were obtained for alkanes. The result shows that the eigenvalue of bonding orbital-connection matrix is a good descriptor for expressing the relative bond energies of C-C and C-H bonds in alkane. This work provides a new physical insight and a principally novel general approach to the evaluation of the bond dissociation energies of carbon-carbon (C-C) and carbon-hydrogen (C-H) bonds in alkane. Also it builds a simple bridge linking the adjacent matrix of radicals R(i)* and R(j)* with the BDE of R(i)-R(j) and R(i)-H. Furthermore, the Heat of Atomization (HA) and Heat of Formation in Gas (HFG(0)) of alkane can be estimated well with the parameters X(1CC), X(1CH), and S(ij). PMID- 12653528 TI - Nanotubes: number of Kekule structures and aromaticity. AB - Carbon nanotubes (CNTs) are composed of cylindrical graphite sheets consisting of sp(2) carbons. Due to their structure CNTs are considered to be aromatic systems. In this work the number of Kekule structures (K) in "armchair" CNTs was estimated by using the transfer matrix technique. All Kekule structures of the cyclic variants of naphthalene and benzo[c]phenanthrene have been generated and the basic patterns have been obtained. From this information the elements of the transfer matrix were derived. The results obtained indicate that K (and the resonance energy) is greater if tubulenes are extended in the vertical than in the horizontal direction. Tubulenes are therefore more stabile than cyclic strips. An illustration, obtained by using scanning probe microscope, has been attached to affirm the existence of thin CNTs. PMID- 12653529 TI - A QSAR model of PAHs carcinogenesis based on thermodynamic stabilities of biactive sites. AB - Polycyclic aromatic hydrocarbons (PAHs) have been adopted to study the carcinogenesis of chemicals experimentally and theoretically. A model of carcinogenic activity of 48 PAHs was obtained based on the calculated relative thermodynamic stabilities of epoxide and carbonium intermediates of the PAHs. Using both epoxyl-energy and cationized-energy of two active sites, the model reasonably predicts the carcinogenic activity of these PAHs and shows a good ability to distinguish between carcinogenic and noncarcinogenic PAHs. Furthermore, the model suggests that double active sites and their distance characteristics are important factors in the chemical carcinogenesis of PAHs. The physical meaning of the energies corresponding addition reactions with DNA is also discussed. PMID- 12653530 TI - An atlas of forecasted molecular data. 1. Internuclear separations of main-group and transition-metal neutral gas-phase diatomic molecules in the ground state. AB - Needed spectroscopic data on diatomic molecules can often be found in the superb critical tables of Huber and Herzberg or in the literature published since 1979. Unfortunately, these sources apply to only a fraction of the diatomic species that can exist and so investigators have had to rely on interpolation, additivity, or ad hoc rules to estimate needed values, all of which require other information that is often lacking. This Atlas presents 1001 additional internuclear separations for use until critical tables are available to fill the needs more precisely. The Atlas was produced by mining the data from Huber and Herzberg for trends with least-squares analysis and with neural network software. There are 162 molecules about whose data Huber and Herzberg had no qualifications and whose data were employed for this work; 248 copies of data with low and high magnitudes were added to reduce the effects of frequency. Internuclear separations for 1001 species not found in Huber and Herzberg are presented, and least-squares predictions supplement some of them. The results, i.e., the Atlas, are presented as Table A, Supporting Information. The average error, based on the average of the absolute differences between the predicted values and tabulated values for the molecules having Huber and Herzberg data, is 0.074 A; if each error is expressed as a percent of the forecast to which it pertains, the average of these errors is 2.94%. There are 25 "questionable" data from Huber and Herzberg, not used in the preparation of the Atlas, for which predictions are included in the Atlas. Of these, 14 agree with the predicted internuclear separations to within twice the stated errors. Additional atlases for other properties of diatomic molecules are in preparation. PMID- 12653532 TI - A QSPR study of O-H bond dissociation energy in phenols. AB - A Quantitative Structure-Property Relationship (QSPR) is developed for the O-H bond dissociation energy (BDE) of a set of 78 phenols. The data set was composed of monosubstituted, disubstituted, and polysubstituted phenolic derivatives containing substituents with different steric and electronic effects in the ortho , meta-, and para-positions of the aromatic ring. The proposed model, derived from multiple linear regression, contains seven descriptors calculated solely from the molecular structure of compounds. The average absolute relative errors are 1.37% (R(2) = 0.8978; SD: 6.67) and 1.13% (R(2) = 0.9076; SD: 4.26) for the working set (62 compounds) and the prediction set (16 compounds), respectively. These results are better than those obtained from DFT calculations, QSAR approach, and correlations with Hammet parameters. PMID- 12653531 TI - Validation of DAPPER for 3D QSAR: conformational search and chirality metric. AB - Adequate conformational searching of small molecules and inclusion of a chirality identifier are necessary features of any current technique for quantitative structure-activity relationships (QSAR). However, implementation of these features can be difficult and computationally expensive, and some techniques can still lead to insufficient treatment of molecular conformation. We select the standard systematic conformational search as the default search method for our recent 3D QSAR program, DAPPER, and develop a novel chirality metric for use in QSAR. These techniques are implemented in DAPPER and validated on standard data sets. PMID- 12653533 TI - Correlation between the glass transition temperatures and repeating unit structure for high molecular weight polymers. AB - A set of five-parameter descriptors, sum MV(ter)(R(ter)), L(F), DeltaX(SB), sum PEI, and Q(+/-), are developed to express the chain stiffness (or mobility) and the intermolecular forces of polymers. Investigated results show a good correlation (R = 0.9517, R(2) = 0.9056, s = 20.86 K) between the glass transition temperatures (T(g)s) and the five parameters for a diverse set of 88 polymers. The descriptors are easy to calculate directly from the repeating unit structure and have clear physical meanings. This approach provides a new insight for Quantitative Structure-Property Relationship (QSPR) correlation of glass transition temperatures of high molecular weight polymers. PMID- 12653534 TI - Pharmacophore modeling as an efficient tool in the discovery of novel noncompetitive AMPA receptor antagonists. AB - A three-dimensional pharmacophore model for the binding of noncompetitive AMPA receptor antagonists was developed in order to map common structural features of highly active compounds. This hypothesis, which consists of two hydrophobic regions, one hydrogen bond acceptor and one aromatic region, was successfully used as framework for the design of a new class of allosteric modulators containing a tetrahydroisoquinoline skeleton and for in silico screening. The promising biological results suggested that the identified molecules might be useful "lead compounds" for future drug development. PMID- 12653535 TI - The comparative molecular surface analysis (CoMSA) with modified uniformative variable elimination-PLS (UVE-PLS) method: application to the steroids binding the aromatase enzyme. AB - The application of the CoMSA method to analyze 3D QSAR of 50 steroid aromatase inhibitors is described. The 3D QSAR model obtained, reaching a value of cross validated q(2) = 0.96 (s = 0.31), significantly outperforms those reported in the literature for the CoMFA or CoSA (CoSASA). It is shown that the Uniformative Variable Elimination UVE-PLS or modified iterative UVE procedure (IVE-PLS) can be used for indicating the regions contributing to the binding activity. Thus, after separating the series into two groups of the training and test molecules quite correct external predictions result from the processing of the training set. We proved that the procedure of the data elimination provides stable results, if tested in 50 random runs of the IVE-PLS-CoMSA with different training/test sets. Depending upon the procedure used the quality of the predictions for 25 test molecules is given by SDEP = sum(y(pred)-y(obs))(2)/n)(1/2) = 0.321 - 0.782. PMID- 12653536 TI - Active learning with support vector machines in the drug discovery process. AB - We investigate the following data mining problem from computer-aided drug design: From a large collection of compounds, find those that bind to a target molecule in as few iterations of biochemical testing as possible. In each iteration a comparatively small batch of compounds is screened for binding activity toward this target. We employed the so-called "active learning paradigm" from Machine Learning for selecting the successive batches. Our main selection strategy is based on the maximum margin hyperplane-generated by "Support Vector Machines". This hyperplane separates the current set of active from the inactive compounds and has the largest possible distance from any labeled compound. We perform a thorough comparative study of various other selection strategies on data sets provided by DuPont Pharmaceuticals and show that the strategies based on the maximum margin hyperplane clearly outperform the simpler ones. PMID- 12653537 TI - A consensus neural network-based technique for discriminating soluble and poorly soluble compounds. AB - BCUT [Burden, CAS, and University of Texas] descriptors, defined as eigenvalues of modified connectivity matrices, have traditionally been applied to drug design tasks such as defining receptor relevant subspaces to assist in compound selections. In this paper we present studies of consensus neural networks trained on BCUTs to discriminate compounds with poor aqueous solubility from those with reasonable solubility. This level was set at 0.1 mg/mL on advice from drug formulation and drug discovery scientists. By applying strict criteria to the insolubility predictions, approximately 95% of compounds are classified correctly. For compounds whose predictions have a lower level of confidence, further parameters are examined in order to flag those considered to possess unsuitable biopharmaceutical and physicochemical properties. This approach is not designed to be applied in isolation but is intended to be used as a filter in the selection of screening candidates, compound purchases, and the application of synthetic priorities to combinatorial libraries. PMID- 12653538 TI - Neighborhood behavior of in silico structural spaces with respect to in vitro activity spaces-a novel understanding of the molecular similarity principle in the context of multiple receptor binding profiles. AB - As a consequence of recent advances in the field of High Throughput Screening, the systematic testing ("in vitro profiling") of compounds against a panel of targets covering different therapeutic areas is nowadays used to generate relevant information with respect to the in vivo behavior of drug candidates. However, the development of chemoinformatics tools required for the exploitation of such data is yet in an incipient phase. In this paper, a formalism for the analysis of activity profile vectors (describing the experimental responses of compounds in each of the considered activity tests) is introduced and applied at the study of Neighborhood Behavior (NB; the hypothesis that structurally similar compounds display similar biological properties) of molecular similarity metrics. The experimental activity profiles define an Activity Space in which more than 500 drugs and reference compounds are positioned, their coordinates being inhibitory propensities in the included tests and unambiguously characterizing a molecule in terms of its receptor binding properties. While previous studies of Neighborhood Behavior had to rely on a loose classification of compounds in terms of the therapeutic areas they were designed for, here the NB of a calculated "in silico" similarity metric has been redefined as a relationships between intermolecular dissimilarity scores in the "structural" and "activity" spaces, respectively, and expressed in terms of two quantitative criteria: "consistency" (the propensity of the metric to selectively rank activity-related compound pairs among the structurally most similar pairs) and "completeness" (monitoring the retrieval rate of activity-related compound pairs among the best ranked pairs of structural neighbors). These criteria were used to calibrate and validate a similarity metric based on Fuzzy Bipolar Pharmacophore Fingerprints. PMID- 12653539 TI - Neighborhood behavior of in silico structural spaces with respect to in vitro activity spaces--a benchmark for neighborhood behavior assessment of different in silico similarity metrics. AB - In a previous work, we have introduced Neighborhood Behavior (NB) criteria for calculated molecular similarity metrics, based on the analysis of in vitro activity spaces that simultaneously monitor the responses of a compound with respect to an entire panel of biologically relevant receptors and enzymes. Now, these novel NB criteria will be used as a benchmark for the comparison of different in silico molecular similarity metrics, addressing the following topics: (1) the relative performance of 2D vs 3D descriptors, (2) the importance of the similarity scoring function for a given descriptor set, and (3) binary or Fuzzy Pharmacophore Fingerprints-can they capture the similarity of the spatial distribution of pharmacophoric groups despite different molecular connectivity? It was found that fuzzy pharmacophore descriptors (FBPA) displayed an optimal NB and, unlike their binary counterparts, were successful in evidencing pharmacophore pattern similarity independently of topological similarity. Topological FBPA, identical to the former except for the use of topological instead of 3D atom pair distances, display a somehow weaker, but significant NB. Metrics based on "classical" global 2D and 3D molecular descriptors and a Dice scoring function also performed well. The choice of the similarity scoring function is therefore as important as the choice of the appropriate molecular descriptors. PMID- 12653540 TI - A structure-activity study of taxol, taxotere, and derivatives using the electronic indices methodology (EIM). AB - Among the new families of effective anticancer drugs, the natural product paclitaxel (Taxol/Bristol-Myers-Squibb) and its semisynthetic derivative docetaxel (Taxotere/Rhone-Poulenc Rorer) are probably the most promising agents under investigation. Surprisingly considering their importance no detailed quantum mechanical studies have been carried out for these drugs. In this work we report the first structure--activity relationship (SAR) studies for 20 taxoid structures using molecular descriptors from all-electron quantum methods. The used methods were the pattern-recognition Principal Component Analysis (PCA), Hierarchical Clustering Analysis (HCA), and the recently developed Electronic Indices Methodology (EIM). The combined use of EIM with PCA/HCA methodologies was able to correctly classify active and inactive taxoids with 100% of accuracy using only a few "universal" quantum molecular descriptors. It was possible to identify the electronic features defining active molecules. This information can be used to select and design new active compounds. The combined use of EIM with PCA/HCA can be a new and very efficient tool in the field of computer assisted drug design. PMID- 12653541 TI - Primary reactions of the LOV2 domain of phototropin, a plant blue-light photoreceptor. AB - The phototropins constitute an important class of plant photoreceptor kinases that control a range of physiological responses, including phototropism, light directed chloroplast movement, and light-induced stomatal opening. The LOV2 domain of phototropin binds a molecule of flavin mononucleotide (FMN) and undergoes a photocycle involving light-driven covalent adduct formation between a conserved cysteine residue and the C(4a) atom of FMN. This product state promotes C-terminal kinase activation and downstream signal transduction. Here, we report the primary photophysics and photochemistry of LOV2 domains of phototropin 1 of Avena sativa (oat) and of the phy3 photoreceptor of Adiantum capillus-veneris (maidenhair fern). In agreement with earlier reports [Swartz, T. E., et al. (2001) J. Biol. Chem. 276, 36493-36500], we find that the FMN triplet state is the reactive species from which the photoreaction occurs. We demonstrate that the triplet state is the primary photoproduct in the LOV2 photocycle, generated at 60% efficiency. No spectroscopically distinguishable intermediates precede the FMN triplet on the femtosecond to nanosecond time scale, indicating that it is formed directly via intersystem crossing (ISC) from the singlet state. Our results indicate that the majority of the FMN triplets in the LOV2 domain exist in the protonated form. We propose a reaction mechanism that involves excited state proton transfer, on the nanosecond time scale or faster, from the sulfhydryl group of the conserved cysteine to the N5 atom of FMN. This event promotes adduct formation by increasing the electrophilicity of C(4a) and subsequent nucleophilic attack by the cysteine's thiolate anion. Comparison to free FMN in solution shows that the protein environment of LOV2 increases the ISC rate of FMN by a factor of 2.4, thus improving the yield of the cysteinyl-flavin adduct and the efficiency of phototropin-mediated signaling processes. PMID- 12653542 TI - Quinolone action against human topoisomerase IIalpha: stimulation of enzyme mediated double-stranded DNA cleavage. AB - Several important antineoplastic drugs kill cells by increasing levels of topoisomerase II-mediated DNA breaks. These compounds act by two distinct mechanisms. Agents such as etoposide inhibit the ability of topoisomerase II to ligate enzyme-linked DNA breaks. Conversely, compounds such as quinolones have little effect on ligation and are believed to stimulate the forward rate of topoisomerase II-mediated DNA cleavage. The fact that there are two scissile bonds per double-stranded DNA break implies that there are two sites for drug action in every enzyme-DNA cleavage complex. However, since agents in the latter group are believed to act by locally perturbing DNA structure, it is possible that quinolone interactions at a single scissile bond are sufficient to distort both strands of the double helix and generate an enzyme-mediated double-stranded DNA break. Therefore, an oligonucleotide system was established to further define the actions of topoisomerase II-targeted drugs that stimulate the forward rate of DNA cleavage. Results indicate that the presence of the quinolone CP-115,953 at one scissile bond increased the extent of enzyme-mediated scission at the opposite scissile bond and was sufficient to stimulate the formation of a double stranded DNA break by human topoisomerase IIalpha. These findings stand in marked contrast to those for etoposide, which must be present at both scissile bonds to stabilize a double-stranded DNA break [Bromberg, K. D., et al. (2003) J. Biol. Chem. 278, 7406-7412]. Moreover, they underscore important mechanistic differences between drugs that enhance DNA cleavage and those that inhibit ligation. PMID- 12653543 TI - Structural characterization of yeast acidic ribosomal P proteins forming the P1A P2B heterocomplex. AB - Acidic ribosomal P proteins form a distinct lateral protuberance on the 60S ribosomal subunit. In yeast, this structure is composed of two heterocomplexes (P1A-P2B and P1B-P2A) attached to the ribosome with the aid of the P0 protein. In solution, the isolated P proteins P1A and P2B have a flexible structure with some characteristics of a molten globule [Zurdo, J., et al. (2000) Biochemistry 39, 8935-8943]. In this report, the structure of P1A-P2B heterocomplex from Saccharomyces cerevisiae is investigated by means of size-exclusion chromatography, chemical cross-linking, circular dichroism, light scattering, and fluorescence spectroscopy. The circular dichroism experiment shows that the complex could be ranked in the tertiary class of all-alpha proteins, with an average alpha-helical content of approximately 65%. Heat and urea denaturation experiments reveal that the P1A-P2B complex, unlike the isolated proteins, has a full cooperative transition which can be fitted into a two-state folding unfolding model. The behavior of the complex in the presence of 2,2,2 trifluoroethanol also resembles a two-state folding-unfolding transition, further supporting the idea that the heterocomplex contains well-packed side chains. In conclusion, the P1A-P2B heterocomplex, unlike the isolated proteins, has a well defined hydrophobic core. Consequently, the complex can put up its structure without additional ribosomal components, so the heterodimeric complex reflects the intrinsic properties of the two analyzed proteins, indicating thus that this is the only possible configuration of the P1A and P2B proteins on the ribosomal stalk structure. PMID- 12653544 TI - A group II intron inserted into a bacterial heat-shock operon shows autocatalytic activity and unusual thermostability. AB - Group II intron RNAs fold into catalytically active structures that catalyze their own self-splicing and subsequent transposition into DNA. Because of their remarkable enzymatic properties, it has been of interest to find new group II introns with novel properties. Here we report the cloning, sequencing, and mechanistic characterization of a new group II intron from the bacterium Azotobacter vinelandii (the AV intron). Although it bears the characteristics of the group IIB1 class, the AV intron is unusually G-C rich, and it has unusual insertion sequences and a minimal dependence on the EBS2-IBS2 tertiary interaction. The AV intron is the first bacterial intron that has been found to reside in a housekeeping gene which, in this case, encodes a heat-shock protein (hsp60). Consistent with a potential role in heat-shock regulation, kinetic analysis reveals that AV intron self-splicing is activated only at elevated temperatures. This suggests a novel pathway for the regulation of heat shock in prokaryotes and provides a first example of a thermally tolerant group II intron RNA. PMID- 12653545 TI - Activation of phenylalanine hydroxylase: effect of substitutions at Arg68 and Cys237. AB - Phenylalanine hydroxylase (PAH) is a multidomain tetrameric enzyme that displays positive cooperative substrate binding. This cooperative response is believed to be of physiological significance as a mechanism that controls L-Phe homeostasis in blood. The substrate induces an activating conformational change in the enzyme affecting the secondary, tertiary, and quaternary structures. Chemical modification and substitution with a negatively charged residue of Cys237 in human PAH (hPAH) also result in activation of the enzyme. As seen in the modeled structure of full-length hPAH, Cys237 is located in the catalytic domain close to residues in the oligomerization and regulatory domains of an adjacent subunit in the dimer, notably to Arg68. This residue is located in a prominent loop (68-75), which also has contacts with the dimerization motif from the same subunit. To investigate further the involvement of Cys237 and Arg68 in the activation of the enzyme, we have prepared mutants of hPAH at these positions, with substitutions of different charge and size. The mutations C237D, R68A, and C237A cause an increase of the basal activity and affinity for L-Phe, while the mutation C237R results in reduced affinity for the substrate and elimination of the positive cooperativity. The conformational changes induced by the mutations were studied by far-UV circular dichroism, fluorescence spectroscopy, and molecular dynamics simulations. All together, our results indicate that the activating mutations induce a series of conformational changes including both the displacement of the inhibitory N-terminal sequence (residues 19-33) that covers the active site and the domain movements around the hinge region Arg111-Thr117, in addition to the rearrangement of the loop 68-75. The same conformational changes appear to be involved in the activation of PAH induced by L-Phe. PMID- 12653546 TI - Studies of the local conformational properties of the cell-adhesion domain of collagen type IV in synthetic heterotrimeric peptides. AB - Collagen type IV is a specialized form of collagen that is found only in basement membranes. It is involved in integrin-mediated cell-adhesion processes, and the responsible binding sites for the alpha1beta1 integrin cell receptor have been identified as Asp461 of the two alpha1 chains and Arg461 of the alpha2 chain. In the most plausible stagger of native collagen type IV the alpha2 chain is the tailing one. This has recently been confirmed by the differentiated binding affinities of synthetic heterotrimeric collagen peptides in which the chains were staggered in this native register as well as in the less plausible alpha1alpha2alpha1' register with an artificial cystine knot. In the present work, two heterotrimeric collagen peptides with chain registers identical to the previous ones were synthesized for fluorescence resonance energy transfer and emission anisotropy measurements, exploiting the native Phe464 in the alpha2 chain as donor and an Ile467Tyr mutation in the alpha1' chain as acceptor fluorophore. This fluorophore pair allowed extraction of more detailed information on the conformational properties of the cell-adhesion epitope incorporated into the central part of the trimeric collagen model peptides. A comparison of the experimentally derived values of the interfluorophore distance and of the orientation factor kappa(2) with the values extracted from the molecular model of the trimer in the native stagger confirmed a triple-helical structure of the adhesion-site portion at low temperature. The thermal unfolding of this central domain was specifically monitored by emission anisotropy, allowing unambiguous assignment of the three structural domains of the trimeric collagen molecules detected by microcalorimetry, with the integrin binding site as the portion of weakest triple-helical stability flanked by two more stable triple-helical regions. The results are consistent with the picture of a conformational microheterogeneity as the responsible property for selective recognition of collagens by interacting proteins. PMID- 12653547 TI - Structural roles of subunit cysteines in the folding and assembly of the DNA packaging machine (portal) of bacteriophage P22. AB - The DNA packaging machine (portal assembly) of bacteriophage P22 is constructed from 12 copies of a multidomain 725-residue subunit comprising a complex alpha/beta fold. The portal subunit contains four cysteines (Cys 153, Cys 173, Cys 283, and Cys 516), which produce distinctive Raman markers in the spectral interval 2500-2600 cm(-1) originating from S-H bond-stretching vibrations diagnostic of S-H...X hydrogen-bonding interactions. The Raman spectrum is unique in the capability to characterize cysteine sulfhydryl interactions in proteins and shows that portal cysteine environments are significantly altered by assembly (Rodriguez-Casado et al. (2001) Biochemistry 40, 13583-13591). We have employed site-directed mutagenesis, size-exclusion chromatography, and Raman difference spectroscopy to characterize the roles of portal cysteines in subunit folding and dodecamer assembly. The stability of the portal monomer is severely reduced by a Cys --> Ser point mutation introduced at either residue 173 or 516. In the case of C516S, the destabilized monomer still forms portal rings, as visualized by negative-stain electron microscopy, whereas portal ring formation cannot be detected for C173S, which forms aberrant aggregates. The C283S mutant is a hyperstable monomer that is defective in portal ring formation. Interestingly, Cys 283 is suggested by secondary structure homology with the phi29 portal to be within a domain involved in DNA translocation. Conversely, the phenotype of the C153S mutant is close to that of the wild-type protein, implying that the sulfhydryl moiety of Cys 153 is not essential to formation of the native subunit fold and productive assembly dynamics. The present results demonstrate that cysteines of the P22 portal protein span a wide range of sulfhydryl hydrogen bonding strengths in the wild-type assembly, that three of the four sulfhydryls play key roles in portal protein stability and assembly kinetics, and that substitution of a mutant seryl interaction (O-H...X) for a wild-type cysteinyl interaction (S-H...X) can either stabilize or destabilize the native fold depending upon sequence context. PMID- 12653549 TI - A high-resolution NMR study of long-lived water molecules in both oxidation states of a minimal cytochrome c. AB - The interaction of water with oxidized and reduced cytochrome c from the Gram positive bacterium Bacillus pasteurii (a 71-amino acid long monoheme cytochrome) is investigated through CLEANEX experiments and (15)N-edited ePHOGSY and Tr-ROESY experiments. It appears that a water molecule gives rise to dipolar cross relaxation with the amide protons of Gly74 and Ile75, with a residence time longer than 0.4 ns, to account for a negative NOE. Such water molecule is present in both the oxidized and reduced species and in the X-ray structure. It appears to have a structural role. Other possible roles are discussed by comparison with the water molecules present in other c-type cytochromes. The amide proton of Cys35 is found to exchange rapidly with the solvent in the oxidized but not in the reduced protein, at variance with H/D exchange experiments, which probe a different time scale. The present data confirm that electron-transfer proteins evolved to minimize reorganization energy upon change of the oxidation state, even though the consequent variation of charge of the metal ion may induce some changes in the structure and/or dynamics of the protein. PMID- 12653548 TI - Structural studies on ADP activation of mammalian glutamate dehydrogenase and the evolution of regulation. AB - Glutamate dehydrogenase (GDH) is found in all organisms and catalyzes the reversible oxidative deamination of L-glutamate to 2-oxoglutarate. Unlike GDH from bacteria, mammalian GDH exhibits negative cooperativity with respect to coenzyme, activation by ADP, and inhibition by GTP. Presented here are the structures of apo bovine GDH, bovine GDH complexed with ADP, and the R463A mutant form of human GDH (huGDH) that is insensitive to ADP activation. In the absence of active site ligands, the catalytic cleft is in the open conformation, and the hexamers form long polymers in the crystal cell with more interactions than found in the abortive complex crystals. This is consistent with the fact that ADP promotes aggregation in solution. ADP is shown to bind to the second, inhibitory, NADH site yet causes activation. The beta-phosphates of the bound ADP interact with R459 (R463 in huGDH) on the pivot helix. The structure of the ADP-resistant, R463A mutant of human GDH is identical to native GDH with the exception of the truncated side chain on the pivot helix. Together, these results strongly suggest that ADP activates by facilitating the opening of the catalytic cleft. From alignment of GDH from various sources, it is likely that the antenna evolved in the protista prior to the formation of purine regulatory sites. This suggests that there was some selective advantage of the antenna itself and that animals evolved new functions for GDH through the addition of allosteric regulation. PMID- 12653550 TI - Conservation of electrostatic properties within enzyme families and superfamilies. AB - Electrostatic interactions play a key role in enzyme catalytic function. At long range, electrostatics steer the incoming ligand/substrate to the active site, and at short distances, electrostatics provide the specific local interactions for catalysis. In cases in which electrostatics determine enzyme function, orthologs should share the electrostatic properties to maintain function. Often, electrostatic potential maps are employed to depict how conserved surface electrostatics preserve function. We expand on previous efforts to explain conservation of function, using novel electrostatic sequence and structure analyses of four enzyme families and one enzyme superfamily. We show that the spatial charge distribution is conserved within each family and superfamily. Conversely, phylogenetic analysis of key electrostatic residues provide the evolutionary origins of functionality. PMID- 12653551 TI - Molecular basis of enzyme inactivation by an endogenous electrophile 4-hydroxy-2 nonenal: identification of modification sites in glyceraldehyde-3-phosphate dehydrogenase. AB - 4-Hydroxy-2-nonenal (HNE), a major lipid peroxidation-derived reactive aldehyde, is a potent inhibitor of sulfhydryl enzymes, such as the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). It has been suggested that HNE exerts an inhibitory effect on the enzyme due to the modification of the cysteine residue (Cys-149) at the catalytic site generating the HNE-cysteine Michael addition-type adduct [Uchida, K., and Stadtman, E. R. (1993) J. Biol. Chem. 268, 6388-6393]. In the study presented here, to elucidate the mechanism for the inactivation of GAPDH by HNE, we attempted to identify the modification sites of the enzyme by monitoring the formation of the HNE Michael adducts by mass spectrometric methods. Incubation of GAPDH (1 mg/mL) with 1 mM HNE in 50 mM sodium phosphate buffer (pH 7.4) at 37 degrees C resulted in a time-dependent loss of enzyme activity, which was associated with the covalent binding of HNE to the enzyme. To identify the site of modification of GAPDH by HNE, both the HNE pretreated and untreated GAPDH were digested with trypsin and V8 protease, and the resulting peptides were subjected to electrospray ionization liquid chromatography-mass spectrometry (ESI-LC-MS). This technique identified five peptides, which contained the HNE adducts at His-164, Cys-244, Cys-281, His-327, and Lys-331 and revealed that both His-164 and Cys-281 were very rapidly modified at 5 min, followed by Cys-244 at 15 min and His-327 and Lys-331 at 30 min. These observations and the observation that the HNE modification of the catalytic center, Cys-149, was not observed suggest that the HNE inactivation of GAPDH is not due to the modification of the catalytic center but to the selective modification of amino acids primarily located in the surface of the GAPDH molecule. PMID- 12653552 TI - Energetics of sequence-specific protein-DNA association: binding of integrase Tn916 to its target DNA. AB - The DNA binding domain of the transposon Tn916 integrase (INT-DBD) binds to its DNA target site by positioning the face of a three-stranded antiparallel beta sheet within the major groove. Binding of INT-DBD to a 13 base pair duplex DNA target site was studied by isothermal titration calorimetry, differential scanning calorimetry, thermal melting followed by circular dichroism spectroscopy, and fluorescence spectroscopy. The observed heat capacity change accompanying the association reaction (DeltaC(p)) is temperature-dependent, decreasing from -1.4 kJ K(-1) mol(-1) at 4 degrees C to -2.9 kJ K(-1) mol(-1) at 30 degrees C. The reason is that the partial molar heat capacities of the free protein, the free DNA duplex, and the protein-DNA complex are not changing in parallel when the temperature increases and that thermal motions of the protein and the DNA are restricted in the complex. After correction for this effect, DeltaC(p) is -1.8 kJ K(-1) mol(-1) and temperature-independent. However, this value is still higher than DeltaC(p) of -1.2 kJ K(-1) mol(-1) estimated by semiempirical methods from dehydration of surface area buried at the complex interface. We propose that the discrepancy between the measured and the structure based prediction of binding energetics is caused by incomplete dehydration of polar groups in the complex. In support, we identify cavities at the interface that are large enough to accommodate approximately 10 water molecules. Our results highlight the difficulties of structure-based prediction of DeltaC(p) (and other thermodynamic parameters) and emphasize how important it is to consider changes of thermal motions and soft vibrational modi in protein-DNA association reactions. This requires not only a detailed investigation of the energetics of the complex but also of the folding thermodynamics of the protein and the DNA alone, which are described in the accompanying paper [Milev et al. (2003) Biochemistry 42, 3492-3502]. PMID- 12653553 TI - Energetics of sequence-specific protein-DNA association: conformational stability of the DNA binding domain of integrase Tn916 and its cognate DNA duplex. AB - Sequence-specific DNA recognition by bacterial integrase Tn916 involves structural rearrangements of both the protein and the DNA duplex. Energetic contributions from changes of conformation, thermal motions and soft vibrational modi of the protein, the DNA, and the complex significantly influence the energetic profile of protein-DNA association. Understanding the energetics of such a complicated system requires not only a detailed calorimetric investigation of the association reaction but also of the components in isolation. Here we report on the conformational stability of the integrase Tn916 DNA binding domain and its cognate 13 base pair target DNA duplex. Using a combination of temperature and denaturant induced unfolding experiments, we find that the 74 residue DNA binding domain is compact and unfolds cooperatively with only small deviation from two-state behavior. Scanning calorimetry reveals an increase of the heat capacity of the native protein attributable to increased thermal fluctuations. From the combined calorimetric and spectroscopic experiments, the parameters of protein unfolding are T(m) = 43.8 +/- 0.3 degrees C, DeltaH(m) = 255 +/- 18 kJ mol(-1), DeltaS(m) = 0.80 +/- 0.06 kJ mol(-1), and DeltaC(p) = 5.0 +/- 0.8 kJ K(-1) mol(-1). The DNA target duplex displays a thermodynamic signature typical of short oligonucleotide duplexes: significant heat absorption due to end fraying and twisting precedes cooperative unfolding and dissociation. The parameters for DNA unfolding and dissociation are DeltaH(m) = 335 +/- 4 kJ mol(-1) and DeltaC(p) = 2.7 +/- 0.9 kJ K(-(1) mol(-1). The results reported here have been instrumental in interpreting the thermodynamic features of the association reaction of the integrase with its 13 base pair target DNA duplex reported in the accompanying paper [Milev et al. (2003) Biochemistry 42, 3481 3491]. PMID- 12653554 TI - Phosphorylation of maize and Arabidopsis HMGB proteins by protein kinase CK2alpha. AB - In plants, a variety of chromatin-associated high mobility group (HMG) proteins belonging to the HMGB family have been identified. We have examined the phosphorylation of the HMGB proteins from the monocotyledonous plant maize and the dicotyledonous plant Arabidopsis by protein kinase CK2alpha. Maize CK2alpha phosphorylates the maize HMGB1 and HMGB2/3 proteins and the Arabidopsis HMGB1, HMGB2/3, and HMGB4 proteins. Maize HMGB4 and HMGB5 and Arabidopsis HMGB5 are not phosphorylated by CK2alpha. Depending on the HMGB protein up to five amino acid residues are phosphorylated in the course of the phosphorylation reaction. The HMGB1 proteins from both plants are markedly more slowly phosphorylated by CK2alpha than the other HMGB substrate proteins, indicating that certain HMGB proteins are clearly preferred substrates for CK2alpha. The rate of the phosphorylation reaction appears to be related to the ease of interaction between CK2alpha and the HMGB proteins, as indicated by chemical cross-linking experiments. MALDI/TOF mass spectrometry analyses demonstrate that the HMGB1 and HMGB2/3 proteins occur in various phosphorylation states in immature maize kernels. Thus, HMGB1 exists as monophosphorylated, double-phosphorylated, triple phosphorylated, and tetraphosphorylated protein in kernel tissue, and the tetraphosphorylated form is the most abundant version. The observed in vivo phosphorylation states indicate that protein kinase(s) other than CK2alpha contribute(s) to the modification of the plant HMGB proteins. The fact that the HMGB proteins are phosphorylated to various extents reveals that the existence of differentially modified forms increases the number of distinct HMGB protein variants in plant chromatin that may be adapted to certain functions. PMID- 12653555 TI - Catalysis by ribonuclease A is limited by the rate of substrate association. AB - The value of k(cat)/K(M) for catalysis of RNA cleavage by ribonuclease (RNase) A can exceed 10(9) M(-1) s(-1) in a solution of low salt concentration. This value approaches that expected for the diffusional encounter of the enzyme and its substrate. To reveal the physicochemical constraints upon catalysis by RNase A, the effects of salt concentration, pH, solvent isotope, and solvent viscosity on catalysis were determined with synthetic substrates that bind to all of the enzymic subsites and thereby enable a meaningful analysis. The pK(a) values determined from pH-k(cat)/K(M) profiles at 0.010, 0.20, and 1.0 M NaCl are inconsistent with the known macroscopic pK(a) values of RNase A. This incongruity indicates that catalysis of RNA cleavage by RNase A is limited by the rate of substrate association, even at 1.0 M NaCl. The effect of solvent isotope and solvent viscosity on catalysis support this conclusion. The data are consistent with a mechanism in which RNase A associates with RNA in an intermediate complex, which is stabilized by Coulombic interactions, prior to the formation of a Michaelis complex. Thus, RNase A has evolved to become an enzyme limited by physics rather than chemistry, a requisite attribute of a perfect catalyst. PMID- 12653556 TI - Polyproline type II conformation in the C-terminal domain of the nuclear pore complex protein gp210. AB - gp210 is a major constituent of the nuclear pore complex (NPC) with possible structural and regulatory roles. It interacts with components of the NPC via its C-terminal domain (CTD), which follows a transmembrane domain and a massive ( approximately 200 kDa) N-terminal region that resides in the lumen of the perinuclear space. Here, we report the solution structure of the human gp210 CTD as determined by various spectroscopic techniques. In water, the CTD adopts an extended, largely unordered conformation, which contains a significant amount of left-handed polyproline type II (PII) helical structure. The conformation of the CTD is altered by high pH, charged detergents, and the hydrogen bond-promoting reagent trifluoroethanol (TFE), which decrease the PII fraction of the fragment. TFE also induces a conformational change in a region containing an SPXX motif whose serine becomes specifically phosphorylated during mitosis. We propose that PII elements in the CTD may play a role in its interaction with the NPC and may serve as recognition sites for regulatory proteins bearing WW or other, unknown PII-binding motifs. PMID- 12653557 TI - Synthesis, enhanced fusogenicity, and solid state NMR measurements of cross linked HIV-1 fusion peptides. AB - In the HIV-1 gp41 and other viral fusion proteins, the minimal oligomerization state is believed to be trimeric with three N-terminal fusion peptides inserting into the membrane in close proximity. Previous studies have demonstrated that the fusion peptide by itself serves as a useful model fusion system, at least to the hemifusion stage in which the viral and target cell lipids are mixed. In the present study, HIV-1 fusion peptides were chemically synthesized and cross-linked at their C-termini to form dimers or trimers. C-terminal trimerization is their likely topology in the fusogenic form of the intact gp41 protein. The fusogenicity of the peptides was then measured in an intervesicle lipid mixing assay, and the assay results were compared to those of the monomer. For monomer, dimer, and trimer at peptide strand/lipid mol ratios between 0.0050 and 0.010, the final extent of lipid mixing for the dimer and trimer was 2-3 times greater than for the monomer. These data suggest that the higher local concentration of peptide strands in the cross-linked peptides enhances fusogenicity and that oligomerization of the fusion peptide in gp41 may enhance the rate of viral/target cell membrane fusion. For gp41, this effect is in addition to the role of the trimeric coiled-coil structure in bringing about apposition of viral and target cell membranes. NMR measurements on the membrane-associated dimeric fusion peptide were consistent with an extended structure at Phe-8, which is the same as has been observed for the membrane-bound monomer in the same lipid composition. PMID- 12653558 TI - Rapid filtration analysis of nucleotide binding to Na,K-ATPase. AB - Transient kinetic analysis of nucleotide binding to pig kidney Na,K-ATPase using a rapid filtration technique shows that the interaction between nucleotide and enzyme apparently follows simple first-order kinetics both for ATP in the absence of Mg(2+) and for ADP in the presence or absence of Mg(2+). Rapid filtration experiments with Na,K-ATPase membrane sheets may nevertheless suffer from a problem of accessibility for a fraction of the ATPase binding sites. Accordingly, we estimate from these data that for ATP binding in the absence of Mg(2+) and the presence of 35 mM Na(+) at pH 7.0 at 20 degrees C, the bimolecular binding rate constant k(on) is about 30 microM(-1) x s(-1) and the dissociation rate constant k(off) is about 8 s(-1). In the presence of 10 mM Mg(2+), the binding rate constant is the same as that in the absence of Mg(2+). For ADP or MgADP the binding rate constant is about 20 microM(-1) x s(-1) and the dissociation rate constant is about 12 s(-1). Results of rapid-mixing stopped-flow experiments with the fluorescent dye eosin are also consistent with a one-step mechanism of binding of eosin to the ATPase nucleotide site. The implication of these results is that nucleotide binding to Na,K-ATPase both in the absence and presence of Mg(2+) appears to be a single-step event, at least on the time scale accessible in these experiments. PMID- 12653560 TI - Interaction of an aromatic dibromoisothiouronium derivative with the Ca(2+) ATPase of skeletal muscle sarcoplasmic reticulum. AB - Isothiouronium compounds [Hoving, S., Bar-Shimon, M., Tijmes, J. J., Goldshleger, R., Tal, D. M., and Karlish, S. J. (1995) J. Biol. Chem. 270, 29788-29793] act as high-affinity competitive antagonists for Na(+) and K(+) (Rb(+)) on the renal Na(+)/K(+)-ATPase where they favor the E1 conformation. We have now characterized the effects of 1,3-dibromo-2,4,6-tris(methylisothiouronium)benzene (Br(2)-TITU) on the Ca(2+)-ATPase of skeletal muscle sarcoplasmic reticulum. Br(2)-TITU inhibited the Ca(2+)-ATPase, both transport and catalytic activity, with a K(0.5) of 5-15 microM. Maximum inhibition was at 10 min with t(0.5) of 3-5 min. Br(2) TITU, 100 microM, quenched Trp autofluorescence by 80%, but the residual signal still responded to Ca(2+) binding. Maximum quenching of fluorescence was at pH 9.0. Total E-P levels, during the steady state of turnover of the Ca(2+)-ATPase, were increased from 0.5 to 5.8 nmol x mg(-1) by Br(2)-TITU at pH 6.8. Trinitrophenyl-ATP (TNP-ATP) superfluorescence, which monitors hydrophobicity of the ATP site, was increased 3-4-fold, suggesting that Br(2)-TITU favors an "E2" like state. Fluorescence was also increased 3-5-fold when E-P was induced with P(i) plus EGTA. Br(2)-TITU increased the rate constants of induction of superfluorescence with ATP plus Ca(2+) from 0.32 to 0.69 s(-1) and with P(i) plus EGTA from 0.84 to 7.45 s(-1). Br(2)-TITU also decreased rate constants for "off" reactions from 2.9 to 0.66 s(-1) and from 10.9 to 0.73 s(-1) for the ATP and P(i) reactions, respectively. Br(2)-TITU, which competitively inhibits the Na(+)/K(+) ATPase, has a novel effect on the Ca(2+)-ATPase. It promotes accumulation of E2-P species due to increased rate of formation and decreased rate of hydrolysis and quenches tryptophan autofluorescence. Br(2)-TITU could be a useful inhibitor to probe intermediate reactions of the Ca(2+)-ATPase that link catalysis with Ca(2+) translocation. PMID- 12653559 TI - Purified human MDR 1 modulates membrane potential in reconstituted proteoliposomes. AB - Human multidrug resistance (hu MDR 1) cDNA was fused to a P. shermanii transcarboxylase biotin acceptor domain (TCBD), and the fusion protein was heterologously overexpressed at high yield in K(+)-uptake deficient Saccharomyces cerevisiae yeast strain 9.3, purified by avidin-biotin chromatography, and reconstituted into proteoliposomes (PLs) formed with Escherichia coli lipid. As measured by pH- dependent ATPase activity, purified, reconstituted, biotinylated MDR-TCBD protein is fully functional. Dodecyl maltoside proved to be the most effective detergent for the membrane solubilization of MDR-TCBD, and various salts were found to significantly affect reconstitution into PLs. After extensive analysis, we find that purified reconstituted MDR-TCBD protein does not catalyze measurable H(+) pumping in the presence of ATP. In the presence of physiologic [ATP], K(+)/Na(+) diffusion potentials monitored by either anionic oxonol or cationic carbocyanine are easily established upon addition of valinomycin to either control or MDR-TCBD PLs. However, in the absence of ATP, although control PLs still maintain easily measurable K(+)/Na(+) diffusion potentials upon addition of valinomycin, MDR-TCBD PLs do not. Dissipation of potential by MDR TCBD is clearly [ATP] dependent and also appears to be Cl(-) dependent, since replacing Cl(-) with equimolar glutamate restores the ability of MDR-TCBD PLs to form a membrane potential in the absence of physiologic [ATP]. The data are difficult to reconcile with models that might propose ATP-catalyzed "pumping" of the fluorescent probes we use and are more consistent with electrically passive anion transport via MDR-TCBD protein, but only at low [ATP]. These observations may help to resolve the confusing array of data related to putative ion transport by hu MDR 1 protein. PMID- 12653561 TI - Regulation of c-Fes tyrosine kinase activity by coiled-coil and SH2 domains: analysis with Saccharomyces cerevisiae. AB - The c-Fes protein-tyrosine kinase regulates the growth and differentiation of diverse cell types, including myeloid hematopoietic cells, vascular endothelial cells, and neurons. Structurally, Fes is composed of a unique N-terminal region with coiled-coil oligomerization motifs, followed by SH2 and kinase domains. Although Fes kinase activity is tightly regulated in cells, the structural basis for its negative regulation is not clear. In this report, c-Fes was expressed in Saccharomyces cerevisiae to determine whether regulation is kinase-intrinsic or dependent upon protein factors found in mammalian cells. Wild-type Fes kinase activity was completely repressed in yeast and did not affect cell growth. Mutation or deletion of the more N-terminal c-Fes coiled-coil domain reversed negative regulation, leading to strong kinase activation and suppression of yeast cell growth. Similarly, replacement of the wild-type SH2 domain with that of v Src induced strong kinase activation and the growth-inhibitory phenotype. Immunoblotting with phosphospecific antibodies shows that activation of Fes by either mechanism induced autophosphorylation of the activation loop tyrosine residue (Tyr 713). These data support the idea that Fes naturally adopts an inactive conformation in vivo, and that maintenance of the inactive structure requires the coiled-coil and SH2 domains. PMID- 12653562 TI - During apoptosis of tumor cells HMGA1a protein undergoes methylation: identification of the modification site by mass spectrometry. AB - Programmed cell death is characterized by posttranslational modifications of a limited and specific set of nuclear proteins. We demonstrate that during apoptosis of different types of tumor cells there is a monomethylation of the nuclear protein HMGA1a that is associated to its previously described hyperphosphorylation/dephosphorylation process. HMGA1a methylation is strictly related to the execution of programmed cell death and is a massive event that involves large amounts of the protein. In some tumor cells, HMGA1a protein is already methylated to an extent that depends on cell type. The degree of methylation in any case definitely increases during apoptosis. In the studied cell systems (human leukaemia, human prostate tumor, and rat thyroid transformed cells) among the low-molecular-mass HMG proteins, only HMGA1a was found to be methylated. A tryptic digestion map of HPLC-purified HMGA1a protein showed that methylation occurs at arginine 25 in the consensus G(24)R(25)G(26) that belongs to one of the DNA-binding AT-hooks of the protein. An increase of HMGA1a methylation could be related to heterochromatin and chromatin remodeling of apoptotic cells. PMID- 12653563 TI - Solid-state NMR investigation of the buried X-proline peptide bonds of bacteriorhodopsin. AB - The role of proline residues in the photocycle of bacteriorhodopsin (bR) is addressed using solid-state NMR. (13)C and (15)N chemical shifts from X-Pro peptide bonds in bR are assigned from REDOR difference spectra of pairwise labeled samples, and correlations of chemical shifts with structure are explored in a series of X-Pro model compounds. Results for the three membrane-embedded X Pro bonds of bR indicate only slight changes in the transition from the resting state of the protein to either the early or late M state of the protonmotive photocycle. These results suggest that the buried prolines serve a principally structural role in bR. PMID- 12653564 TI - Characterization of the metal ion-binding domains from rat alpha- and beta parvalbumins. AB - We have examined the metal ion-binding domains from rat alpha and beta parvalbumin. We find that the CD-EF fragments differ markedly in their tendency to self-associate. Whereas Ca(2+)-free alpha CD-EF is monomeric, the Ca(2+)-free beta peptide dimerizes weakly (K(2) = 2400 +/- 200 M(-1)). In buffer containing 1.0 mM Ca(2+), the apparent dimerization constant for beta CD-EF (191,000 +/- 29,000 M(-1)) is more than 50 times that of alpha (3400 +/- 200 M(-1)). Alpha CD EF binds two Ca(2+) with positive cooperativity. Titration calorimetry data afford binding constants of 3.7(0.1) x 10(3) M(-1) and 8.6(0.2) x 10(4) M(-1). Beta CD-EF also binds two Ca(2+) cooperatively but with lower affinity. Equilibrium dialysis yields Adair constants of 4.2(0.1) x 10(3) and 6.1(0.2) x 10(3) M(-1). Significantly, the difference in Ca(2+) affinity is substantially smaller than that observed for the full-length proteins-suggesting that the AB domain can modulate divalent ion affinity. Analysis of beta calorimetry data requires explicit consideration of the self-association behavior. Data collected at low CD-EF concentration are consistent with preferential occupation of the EF site, dimerization of singly bound monomers, and cooperative filling of the CD sites. At higher concentrations, apo-protein dimerization can apparently precede cooperative occupation of the EF sites. In the presence of Ca(2+), alpha CD-EF exhibits higher thermal stability, consistent with its higher Ca(2+) affinity. However, the beta melting temperature shows greater concentration dependence, consistent with its greater tendency to dimerize. Neither fragment exhibits a sigmoidal melting curve in the Ca(2+)-free state, suggesting that the apo peptides are disordered. PMID- 12653565 TI - Stability, miscoding potential, and repair of 2'-deoxyxanthosine in DNA: implications for nitric oxide-induced mutagenesis. AB - Nitric oxide (NO(*)) reacts with guanine in DNA and RNA to produce xanthine (X) as a major product. Despite its potential importance in NO(*)-mediated mutagenesis, the biochemical properties of X in polynucleotides have been relatively unexplored. We describe the synthesis and chemical characterization of xanthine-containing oligonucleotides and report on the susceptibility of X to depurination, its miscoding potential during replication by polymerases, and its recognition and excision by several members of the base excision repair (BER) family of DNA glycosylases. At neutral pH, X was found to be only slightly less stable than guanine to depurination (k(X)/k(G) = 1.19), whereas at pH Mpg > Nth > Fpg. Implications of these results for the induction of mutations by nitric oxide are discussed. PMID- 12653566 TI - pH-induced alteration and oxidative destruction of heme in purified chromaffin granule cytochrome b(561): implications for the oxidative stress in catecholaminergic neurons. AB - The transmembrane hemoprotein, cytochrome b(561) (b(561)), in the neuroendocrine secretory vesicles is shown to shuttle electrons from the cytosolic ascorbate (Asc) to the intravesicular matrix to provide reducing equivalents for the dopamine beta-monooxygenase (DbetaM) reaction. Intravesicular Asc may also play a role in relieving catecholamine-induced oxidative stress in catecholaminergic neurons. In the present study, we have examined the alteration of purified oxidized b(561) (b(561,ox)) under mild alkaline conditions to probe the structural and functional characteristics of the protein, using UV-vis and EPR spectroscopic and kinetic techniques. Our results show that low spin heme in oxidized b(561) (b(561,ox)) readily transforms to an altered high spin form and then slowly to an Asc nonreducible form, in a pH-, temperature-, and time dependent manner, which can be described by single-exponential rate equations, A(t) = A(o)(1- e (-kt)) and A(t) = A(o)e(-kt), respectively. More than half of the Asc nonreducible altered b(561) could be converted back to the native b(561) by pH adjustment followed by dithionite reduction, suggesting the reversibility of the process. The heme center of the transformed Asc nonreducible protein is completely bleached instantaneously by dithionite in the presence of atmospheric oxygen, which appears to be mediated by molecular oxygen and/or hydrogen peroxide. These results demonstrate that the heme centers of the protein are susceptible to the pH-induced alteration and oxidative destruction, raising some questions regarding the proposed one alkaline labile, two-heme model of b(561) [Tsubaki, M.; Nakayama, M.; Okuyama, E.; Ichikawa, Y. (1997) J. Biol. Chem. 272, 23206-23210]. The pH-induced alteration and the destruction of heme under oxidative conditions may play a significant role in the amplification of oxidative stress in catecholaminergic neurons. PMID- 12653567 TI - The transcriptional activity of the APP intracellular domain-Fe65 complex is inhibited by activation of the NF-kappaB pathway. AB - The beta-amyloid precursor protein (APP) is an integral membrane protein that is the subject of proteolytic processing. Sequential cleavage of APP by beta secretase and subsequently gamma-secretase generates the beta-amyloid peptide as well as a cytoplasmic intracellular domain (AICD). AICD binds to the transcriptional coactivator Fe65, and this complex has been shown to display transcriptional activity. The regulation of this complex is poorly understood. We show here that activation of the NF-kappaB pathway, either by overexpression of NF-kappaB-inducing kinase (NIK) or by treatment with the proinflammatory cytokine IL-1beta, downregulates the transcriptional activity of the AICD-Fe65 complex. This therefore provides a mechanism by which the activity of AICD might be modulated by extracellular stimuli. These results also identify an intracellular signal transduction pathway influenced by the NF-kappaB signaling pathway. PMID- 12653568 TI - Primary salivary clear cell tumors. PMID- 12653570 TI - A longitudinal replicate study of immunosuppressive drugs: a College of American Pathologists study. AB - OBJECTIVE: To identify the sources of analytical variation for cyclosporine and tacrolimus in a 3-year longitudinal study. DESIGN: Two pools of whole blood were spiked with cyclosporine and tacrolimus, respectively. One aliquot of cyclosporine and 2 of the tacrolimus pool were distributed in the first and last mailing for years 1999 to 2001. For both drugs, the total variance for each method was partitioned into within- and between-laboratory components. SETTING: The A and C mailings of the 1999, 2000, and 2001 AACC/CAP [American Association for Clinical Chemistry/College of American Pathologists] Immunosuppressive Drugs (CS) Monitoring Survey. MAIN OUTCOME MEASURES: For each drug, total variance was partitioned into specimen, mailing, year, and interlaboratory effects for each analytical method. PARTICIPANTS: The 292 laboratories for cyclosporine and 177 laboratories for tacrolimus enrolled in the survey from 1999 to 2001. RESULTS: For both cyclosporine and tacrolimus, the major source of imprecision came from within-laboratory factors, which accounted for nearly 85% (range, 77% to 90%) of the total variance. For cyclosporine, the major component of within-laboratory variance was between-mailing, within-year effect, whereas for tacrolimus it was the between-year, within-laboratory variation. CONCLUSION: The major source of long-term survey imprecision for cyclosporine and tacrolimus is within-laboratory factors. The finding that 85% of the total variance was due to within-laboratory variation is similar to other therapeutic drugs. PMID- 12653571 TI - Reactivity of resident immunocytes in normal and prepsoriatic skin using an ex vivo skin-explant model system. AB - CONTEXT: While it is well known that both exogenous and endogenous stimuli can trigger appearance of psoriatic lesions, the initial cellular and molecular events mediated by immunocompetent cells normally resident in prepsoriatic (PN) skin are not well understood. Moreover, it is unclear whether there are any fundamentally important differences in the innate immune response of normal healthy skin (NN skin) versus PN skin. Since acute tissue responses to stimuli involve both resident cells and immunocytes recruited rapidly from circulation, it is difficult to discern the contribution of endogenous cells normally present in skin. OBJECTIVE: To solely characterize the reactivity of resident immunocytes using an experimental system. DESIGN: To probe the activation potential of resident immunocytes in NN (n = 18) and PN skin (n = 10), a short-term ex vivo organ culture system containing interleukin (IL)-2 was established and characterized. To mimic exogenous or environmental trigger factors, bacteria derived superantigens and lipopolysaccharide were added to the skin-explant assays, whereas endogenous trigger factors were investigated using heat shock proteins. RESULTS: Using this skin-explant assay, both NN and PN skin gave rise to an expansion of various T-cell subsets, which could differentially produce various cytokines and a growth factor (keratinocyte growth factor), depending on the stimulus and source of skin. Bacterial superantigens were relatively potent inducers of interferon-gamma, and natural killer-T cells were observed proliferating from PN skin. CONCLUSIONS: Despite relatively few T cells normally residing in either NN or PN skin, initiation of skin explants from both sets of individuals in the presence of IL-2 triggered vigorous T-cell proliferation and cytokine/growth factor release. These results demonstrate the utility of this skin-explant assay system to further investigate quantitative and qualitative immune responses of NN and PN skin. PMID- 12653572 TI - Sinonasal smooth muscle cell tumors: a clinicopathologic and immunohistochemical analysis of 12 cases with emphasis on the low-grade end of the spectrum. AB - CONTEXT: Smooth muscle tumors (SMTs) of the sinonasal tracts are very rare mesenchymal neoplasms, and the literature includes very limited data correlating histologic parameters with clinical outcome. As the behavior of SMTs is site dependent, defining applicable criteria to distinguish among low-grade leiomyosarcoma, SMT of uncertain malignant potential (SMTUMP), and cellular leiomyoma is sometimes difficult and arbitrary. OBJECTIVE: To correlate the clinicopathologic features of 12 well-differentiated sinonasal SMTs with MIB-1 index and clinical outcomes so as to better classify this group of tumors. DESIGN: Twelve cases of sporadic well-differentiated SMTs arising from the sinonasal tract were retrieved from both institutions. High-grade leiomyosarcomas were excluded from this analysis. The histologic parameters assessed included circumscription, mucosal ulceration, cellularity, nuclear atypia, mitotic count, necrosis, and destruction of adjacent bony structures. The histologic classification of these tumors was based on the guidelines for SMTs of deep soft tissues, using greater than 4 mitotic figures (MF)/10 high-power fields (HPF) to separate SMTUMP from leiomyosarcoma and the presence of 1 to 4 MF/10 HPF to distinguish between leiomyoma and SMTUMP. Immunostaining of MIB-1 index was performed in 7 cases with available material. RESULTS: This study showed a 1:1.5 male-female ratio and a mean age of 40 years (range, 20-67 years). The most frequent symptoms were nasal stuffiness and/or epistaxis. The tumors involved nasal cavity in 8 cases (67%), paranasal sinus alone in 2 cases (16.5%), and both nasal cavity and paranasal sinuses in 2 cases (16.5%). The tumors ranged in size from 0.3 to 5.5 cm (mean, 2 cm) and were classified as follows: 7 leiomyomas, 2 SMTUMPs, and 3 low-grade leiomyosarcomas. All 7 leiomyomas, 1 SMTUMP, and 1 leiomyosarcoma originated from vessel walls. Bone involvement was seen in both the SMTUMPs (1 of 2) and leiomyosarcomas (2 of 3). Focal infiltrative growth was observed only in the 3 leiomyosarcomas, 1 of which also showed microscopic coagulative tumor necrosis. Mitotic figures ranged from 0 to 10 (mean, 2.3), with absence of mitosis in all 7 leiomyomas, 1 to 4 MF/10 HPF in the 2 SMTUMPs, and more than 4 MF/10 HPF in the 3 low-grade leiomyosarcomas. The MIB-1 index was low (or=15%) proliferative index. All 12 patients were treated by surgical excision, and only 1 patient with leiomyosarcoma received postoperative radiation. In all 12 tumors, there was neither local recurrence nor distant metastasis after an average of 93 months of follow-up (range, 4-221 months). CONCLUSION: Well-differentiated sinonasal SMTs are rare and occur in adults with a slight female predilection. The lesions confined within the nasal cavity were more common than those involving a single paranasal sinus or both nasal cavity and paranasal sinuses. Despite variations in location, clinical manifestation, histologic features, and MIB-1 index, these tumors appear to follow a favorable course. Complete surgical excision appears to provide adequate treatment for these patients. PMID- 12653573 TI - Mature B-cell leukemias with more than 55% prolymphocytes: report of 2 cases with Burkitt lymphoma-type chromosomal translocations involving c-myc. AB - CONTEXT: The molecular genetic events involved in the pathogenesis of mature B cell leukemias with more than 55% prolymphocytes are not well characterized. We have encountered 2 such cases in which conventional cytogenetic analysis identified Burkitt lymphoma-type chromosomal translocations involving 8q24. OBJECTIVE: To assess these 2 cases for involvement of the c-myc gene using fluorescence in situ hybridization analysis with probes specific for the c-myc and immunoglobulin heavy-chain (IgH) genes. RESULTS: In both cases, conventional cytogenetic analysis demonstrated complex karyotypes, including chromosomal translocations involving 8q24. In case 1, a case of de novo prolymphocytic leukemia, the t(8;14)(q24;q32) was detected. In case 2, a case of chronic lymphocytic leukemia in prolymphocytoid transformation, the t(8;22)(q24;q11) was identified. Fluorescence in situ hybridization studies showed c-myc/IgH fusion signals in case 1, proving the presence of the t(8;14). Split c-myc signals without fusion to IgH were observed in case 2, proving c-myc gene rearrangement and consistent with the t(8;22). CONCLUSION: These results suggest that c-myc gene alterations may be involved in the pathogenesis of a subset of mature B-cell leukemias with more than 55% prolymphocytes. PMID- 12653574 TI - Tissue plasminogen activator plasma levels as a potential diagnostic aid in acute pulmonary embolism. AB - CONTEXT: Pulmonary embolism (PE) is a potentially fatal and frequent complication of deep venous thrombosis, and the most reliable techniques for the diagnosis of PE are not universally available and have other limitations. OBJECTIVE: To determine the efficacy of 4 different fibrinolysis system parameters, namely, tissue plasminogen activator (tPA), tissue plasminogen activator inhibitor type 1 (PAI-1), plasmin-antiplasmin complexes (PAP), and D-dimer, in the diagnosis of acute PE. SETTING: A 350-bed university hospital serving an area with 280,000 inhabitants. PATIENTS: Sixty-six consecutive outpatients with clinically suspected PE. The diagnosis of PE was based on ventilation-perfusion (V/Q) lung scan in combination with clinical assessment, lower limb study, and (when required) pulmonary angiography. MAIN OUTCOME MEASURES: At the moment of clinical suspicion, a sample of venous blood was obtained to measure levels of tPA, PAI-1, PAP, and D-dimer using an enzyme-linked immunosorbent assay method. RESULTS: Twenty-seven patients (41%) were classified as PE positive (high clinical probability and V/Q lung scan [n = 12], nondiagnostic V/Q lung scan and high clinical probability [n = 1], inconclusive V/Q lung scan and positive lower limb examination for deep venous thrombosis [n = 11], and positive pulmonary angiography [n = 3]), and 39 patients (59%) were classified PE negative. The sensitivity/negative predictive value for tPA, using a cutoff of 8.5 ng/mL, and PAI-1, using a cutoff of 15 ng/mL, were 100%/100% and 100%/100%, respectively. A tPA level lower than 8.5 ng/mL occurred in 13 (19.7%; all PE negative) of 66 patients with suspected PE, and PAI-1 levels were lower than 15 ng/mL in 9 (13.6%; all PE negative) of 66 patients with suspected PE. The D-dimer, using a cutoff of 500 ng/mL, showed a sensitivity and negative predictive value of 92.6% and 87.5%, respectively. CONCLUSIONS: Our data indicate that tPA and PAI-1 levels are potentially useful in ruling out PE, although tPA seems to be the better parameter. The sensitivity levels and negative predictive values for the rapid enzyme-linked immunosorbent assay for D-dimer used in this investigation were low compared with previous studies using the same test. PMID- 12653575 TI - Allergic transfusion reactions: an evaluation of 273 consecutive reactions. AB - CONTEXT: Allergic reaction to transfusion is common. However, the review of a large series of allergic transfusion reactions has not been performed. OBJECTIVE: To review a large series of allergic transfusion reactions. DESIGN: A retrospective review of all reported and evaluated transfusion reactions during a 9-year period at 1 institution was performed. Associated clinical signs and symptoms were evaluated. SETTING: Large, tertiary-care teaching hospital. RESULTS: A total of 1613 adverse reactions to transfusion were evaluated. Allergic transfusion reactions accounted for 17% (273 of 1613) of the transfusion reactions. Severe allergic reactions (anaphylaxis, anaphylactoid signs and symptoms, and/or hypotension) were observed in 21 patients (7.7% of allergic reactions, or 1.3% of all transfusion reactions). Serum tryptase, a marker for anaphylaxis, was measured in 1 patient and determined to be borderline elevated. Five patients experienced allergic transfusion reactions to autologous red cell transfusions. One patient experienced hives during the transfusion of a major ABO mismatched red blood cell. A wide variety of skin manifestations were observed, but 26 (9.5%) patients did not have skin manifestations. Allergic transfusion reactions were estimated to occur in approximately 1 in 4124 blood components transfused, or 1 in 2338 transfusion episodes. Severe allergic reactions occurred in approximately 1 in 30,281 transfusions. No deaths directly attributable to transfusion were observed in this patient group. CONCLUSIONS: The clinical presentation of allergic transfusion reactions was quite variable, and the pathophysiology remains unclear. Recommendations for clinical evaluation and therapy remain problematic and often empirical. PMID- 12653576 TI - Pulmonary sclerosing hemangioma with lymph node metastases: report of 4 cases. AB - CONTEXT: Sclerosing hemangioma is an unusual pulmonary tumor. Previously, 4 patients with pulmonary sclerosing hemangioma and lymph node metastases have been described in the literature. OBJECTIVE: To report 4 additional cases of metastatic sclerosing hemangioma. DESIGN: Retrospective review of the authors' consultation files and review of histologic sections of pulmonary tumors and lymph node metastases. RESULTS: Four cases of a morphologically benign pulmonary sclerosing hemangioma with regional lymph node metastases (including hilar, peribronchial, and interlobar metastases) were identified. The patients (3 female, 1 male) had a mean age of 39 years (range, 10-56 years). The tumors ranged in size (greatest dimension) from 1.5 to 4.7 cm (mean, 3.1 cm). The pulmonary tumors were typical circumscribed sclerosing hemangiomas without mitotic activity, angiolymphatic invasion, or necrosis. One tumor had focal cytologic atypia. The metastases were identified in hilar lymph nodes that were removed at operation for the lung nodule. One patient received adjuvant chemotherapy for adenocarcinoma. All of the patients are alive. No recurrences or residual disease has been detected at a mean follow-up of 4.7 years (range, 2.3 10 years). CONCLUSIONS: On the basis of case data from the 4 patients described here and the 4 patients described previously, metastases to regional lymph nodes from pulmonary sclerosing hemangioma may occur but are rare and do not appear to affect prognosis. PMID- 12653577 TI - Comparison of polymerase chain reaction with histopathologic features for diagnosis of tuberculosis in formalin-fixed, paraffin-embedded histologic specimens. AB - Objective.-To investigate the relationship between various histopathologic features and the results of the tuberculosis (TB)-polymerase chain reaction (PCR) method in routinely submitted histologic specimens for the histopathologic diagnosis of TB. Design.-We used 95 formalin-fixed, paraffin-embedded tissue blocks from 81 patients who were clinically suspected of having TB. We assessed the presence of histopathologic features including well-formed granuloma, poorly formed granuloma, caseous necrosis, and Langhans-type giant cells. We performed nested PCR for IS6110 and Ziehl-Neelsen staining for acid-fast bacilli (AFB). Results.-Of the 81 patients studied, 53 patients had chronic granulomatous inflammation, whereas 28 patients had only chronic inflammation without definite granulomatous inflammation. Of the 53 cases with chronic granulomatous inflammation, 17 (32%) were AFB positive and 36 (68%) were TB-PCR positive. Among cases with chronic granulomatous inflammation, the percentage that were positive and negative by TB-PCR differed significantly with the presence of various histopathologic features. All of the 13 cases with well-formed granuloma, caseous necrosis, and Langhans-type giant cells were TB-PCR positive; however, 10 (36%) of the 28 cases with chronic inflammation without granulomatous lesions were also TB-PCR positive. Conclusions.-TB-PCR is a rapid, sensitive method for the diagnosis of TB in routinely processed formalin-fixed, paraffin-embedded histologic specimens and is readily available in histopathology laboratories. We recommend use of TB-PCR when TB is suspected clinically, especially in cases of chronic inflammation without definite evidence of granulomatous inflammation. PMID- 12653578 TI - Pulmonary expression of vascular endothelial growth factor in sepsis. AB - CONTEXT: Vascular endothelial growth factor (VEGF), an angiogenic and chemotactic peptide, is abundantly expressed in normal lung tissue, especially in alveolar and bronchial epithelium, glandular cells of the bronchi, and activated alveolar macrophages. OBJECTIVE: To investigate the role of VEGF in progressively impaired lung function as the major complication and cause of death in septic patients. MATERIALS AND METHODS: We evaluated pulmonary VEGF expression in lung autopsy material from septic patients who had been cared for by intensive care medicine using enzyme-linked immunosorbent assay (ELISA), reverse transcriptase-polymerase chain reaction (RT-PCR), and immunohistochemical methods. RESULTS: Compared with expression in nonseptic control individuals (n = 10), pulmonary VEGF expression as determined by ELISA was significantly (P <.001) decreased in septic patients (n = 8). As monitored by RT-PCR, mRNA for the 2 splice variants, VEGF(121) and VEGF(165), and for VEGFR-2/KDR were expressed in both groups, the yields being lower in the sepsis group. Samples from septic patients lacked or showed only sparse immunoreaction on bronchial and alveolar epithelium, whereas this reaction was strong in all control samples. However, alveolar macrophages were similarly immunopositive in both groups. CONCLUSIONS: The precise underlying mechanisms for the distinctly different expression of pulmonary VEGF in septic patients and nonseptic control individuals are not clear at present. Particularly the role of VEGF in the development of sepsis-induced lung injury and acute respiratory distress syndrome in mechanically ventilated patients suffering from severe sepsis remains to be clarified. PMID- 12653580 TI - Sinus histiocytosis with massive lymphadenopathy and Langerhans cell histiocytosis express the cellular adhesion molecule CD31. AB - Context.-We investigated expression of the adhesion molecule CD31 in sinus histiocytosis with massive lymphadenopathy (SHML) and Langerhans cell histiocytosis (LCH) because (1) SHML and LCH cells express a variety of cellular adhesion molecules and (2) SHML has been characterized as a reactive histiocytic proliferation, and tissue macrophages (histiocytes) are known to express CD31. Objective.-The purpose of this study was to determine whether SHML and LCH cells express CD31 and whether dual staining with CD31 and S100 facilitates diagnosis of these disease states. Methods.-Formalin-fixed, paraffin-embedded archival tissues were immunohistochemically stained via the labeled streptavidin-biotin method using antibodies against CD31 and S100 protein after heat-induced epitope retrieval. Archival tissues included SHML (n = 2), LCH (n = 10), malignant melanoma (n = 5), sinus hyperplasia (n = 4), granulomas (n = 4), granular cell tumor (n = 6), and normal skin (n = 4). Results.-Normal Langerhans cells in the epidermis were CD31(-)/S100(+); neoplastic Langerhans cells in LCH were CD31(+)/S100(+). Histiocytes in granulomas and in sinus hyperplasia were CD31(+)/S100(-); abnormal histiocytes in SHML were CD31(+)/S100(+). S100(+) tumors (malignant melanoma and granular cell tumor) were CD31(-). Conclusions. The spectrum of cell types that express CD31 is expanded to include SHML and LCH. We speculate that up-regulation of CD31 in neoplastic Langerhans cells contributes to the migratory capability of LCH cells. CD31 may be a useful nonlysosomal marker of macrophages and their neoplastic counterparts (true histiocytic sarcomas). An immunohistochemical staining panel that includes CD31 and S100 facilitates the diagnosis of SHML and LCH. PMID- 12653579 TI - The relationship between p63 and p53 expression in normal and neoplastic breast tissue. AB - CONTEXT: p63 is a recently described p53 homologue. Despite structural homology, they have different activities. OBJECTIVES: To obtain new insights into the role of p63 in normal and neoplastic breast tissue and to verify the possible association between p63 and p53 in breast carcinomas. DESIGN: Immunohistochemistry in 85 breast carcinomas using p63, smooth muscle actin (1A4), p53, estrogen receptor, and progesterone receptor. The p63-positive cases were submitted to a double-immunolabeling study using p63 with 1A4, cytokeratin 7, and 34betaE12. Clinical data were retrieved from medical files. RESULTS: p63, like 1A4, stained a single and continuous layer surrounding normal breast ductal and alveolar epithelium. In carcinomas, p53 was expressed in 21.17% of carcinomas, whereas p63 was expressed only in poorly differentiated ductal carcinomas (11.76% of cases). p63-positive cells coexpressed 1A4 and 34betaE12, but not cytokeratin 7. Expression of p63 correlated with pathologic staging, tumor size, histologic grading, nodal metastasis, and estrogen receptor negativity. CONCLUSIONS: p63 is a specific myoepithelial cell marker in normal breast tissue and is expressed in a minority of breast carcinomas, being seen only in grade III ductal carcinomas. In ductal carcinomas, malignant p63-positive cells have an immunophenotype similar to that of myoepithelial cells, suggesting that these cells originate from a primary progenitor cell that underwent divergent differentiation to ductal and myoepithelial cells during clonal expansion. Our study argues against a direct role in mammary tumorigenesis. However, p53 is rarely coexpressed with p63, suggesting that p63 could act indirectly as an oncogene by inhibiting p53. This hypothesis could also explain why p63 correlated with several other indicators of poor prognosis. PMID- 12653581 TI - Effect of marathon running on total and free serum prostate-specific antigen concentrations. AB - CONTEXT: Prostate-specific antigen (PSA) is an important tumor marker for the most frequently diagnosed cancer in the United States. A major limitation of this marker is falsely elevated results in patients who are found not to have prostate cancer. The effects of vigorous physical exertion on PSA concentrations are controversial. OBJECTIVE: To determine the effects of marathon running on PSA levels. DESIGN: Measurement of total and free PSA levels in the sera of participants in a marathon before and within 4 and 24 hours after the race. RESULTS: None of the participants had elevated total PSA levels before the race. Although we found no statistically significant changes in average total or free PSA concentrations at either time point, after the marathon, 2 (11%) of 18 runners had total PSA concentrations outside the standard reference range. Changes in total PSA levels did not correlate with age or prerace PSA concentrations. Free PSA levels were not statistically significantly changed after the race and did not allow a reliable determination of exercise-induced PSA elevations. CONCLUSIONS: Although it may not be necessary for men to abstain from exercise involving running before blood draws for PSA analysis, elevated PSA concentrations may be observed in some individuals after participation in a major sporting event. In these cases, repeat measurements should be considered at a time significantly removed from such exercise. PMID- 12653582 TI - Posttransplant CD30+ (Ki-1) anaplastic large cell lymphoma: a case report and review of the literature. AB - Posttransplant CD30(+) (Ki-1) anaplastic large cell lymphoma (ALCL) is rare. A review of the literature revealed only 3 such cases. All 3 cases were developed after single-organ transplantation. We describe CD30(+) (Ki-1) ALCL in a dual organ (liver and heart) transplantation recipient. The patient was a 68-year-old white female who underwent an orthotopic heart transplantation in 1999 and a liver transplantation in 2000. She presented with nausea and was found to have CD30(+) (Ki-1) ALCL by pathologic examination of the gastric antrum biopsy specimen. To our knowledge, this patient represents the first reported case of posttransplant CD30(+) ALCL following a dual-organ transplantation. PMID- 12653583 TI - Malignant ameloblastoma, spindle cell variant. AB - In this report, we document the histologic and clinical features of a previously undefined spindle cell variant of ameloblastoma that eventually behaved in a malignant fashion during a protracted course. The predominant histologic pattern was a well-differentiated, cellular, spindled epithelial proliferation arising in the maxilla of a 14-year-old African American girl. Over 19 years, the patient experienced numerous local recurrences, metastases to distant bones after 15 years, and finally bulky local recurrence with intracranial extension resulting in death. This ameloblastic malignancy histologically simulates a low-grade true sarcoma or an ameloblastic sarcoma, but differs in that the extensive spindle cell proliferation is epithelial, characterized by strong cytokeratin immunoreactivity and negative vimentin staining. PMID- 12653585 TI - Angiosarcoma masquerading as embryonal carcinoma in the metastasis from a mature testicular teratoma. AB - Sarcoma can arise within a germ cell tumor (GCT) from a malignant transformation of teratomatous elements or as late sequelae to radiation therapy. Angiosarcoma as a malignant component in testicular GCTs has rarely been reported and is often misdiagnosed as embryonal carcinoma. We report the case of a 23-year-old man with mature teratoma of the testis and retroperitoneal metastasis exhibiting components of mature teratoma intermingled with high-grade angiosarcoma. It is important to recognize the presence of a high-grade sarcomatous component within a GCT because of its aggressive clinical behavior and different response to therapy. PMID- 12653584 TI - Biphenotypic acute leukemia with coexpression of CD79a and markers of myeloid lineage. AB - Acute leukemias demonstrating immunophenotypic features of more than 1 cell lineage are referred to as acute leukemias of ambiguous lineage in the new World Health Organization classification system. A subtype of leukemia of ambiguous lineage is biphenotypic acute leukemia in which the malignant cell population expresses markers of 2 different lineages, most commonly myeloid and either B- or T-lymphoid lineages. This entity has been defined by a scoring system proposed by the European Group for the Immunological Characterization of Acute Leukemias (EGIL), with various markers assigned a score of 2, 1, or 0.5 depending on their specificity for myeloid or lymphoid lineage. Those cases having a score greater than 2 for the myeloid and either the B- or T-lymphoid lineages are biphenotypic acute leukemia in this system. One marker, CD79a, has been so clearly associated with acute lymphoblastic leukemia (ALL) by some researchers that its expression in the presence of blast markers is considered indicative of B-ALL. We describe an unusual case of acute leukemia meeting the criteria for biphenotypic acute leukemia in which CD79a expression was observed in the blast population. PMID- 12653586 TI - Hepatocellular injury with hyperaminotransferasemia after cresol ingestion. AB - A 42-year-old man attempted suicide by ingesting about 150 mL of a saponated cresol solution containing about 50% cresol. His serum aminotransferase concentrations were elevated, and a coagulopathy was present at the time of admission, 15 hours after ingestion. The hyperaminotransferasemia and coagulopathy worsened on the second day, but resolved thereafter with supportive therapy. Histologic examination of a biopsy specimen obtained on the 14th day demonstrated focal dropout of hepatocytes (which were replaced by reticulin and collagen fibers), ballooning or hydropic degeneration of hepatocytes, and rapid regeneration with small hepatocytes in the periportal zones as well as in the centrilobular zones. A rapid onset of illness with periportal hepatocellular injury is inconsistent with damage due to a hepatotoxic metabolite of p-cresol produced by cytochrome P450, which has been suggested by studies in vitro. A direct transient noxious effect mediated via the portal or arterial circulation may be involved in hepatic injury after cresol ingestion. PMID- 12653587 TI - Merkel cell carcinoma: a report of gastrointestinal metastasis and review of the literature. AB - Merkel cell carcinoma (MCC) is an uncommon, highly aggressive cutaneous neoplasm of neuroendocrine differentiation with a poor prognosis. MCC most often presents as a painless, firm, raised lesion in sun-exposed sites of the head and neck region of the elderly. We report a case of a metastatic MCC to the stomach presenting as upper gastrointestinal bleeding. To our knowledge, this is the second reported case of MCC presenting as upper gastrointestinal bleeding and the first case confirmed by the newer immunohistochemical techniques. The literature is reviewed. PMID- 12653588 TI - Primary cutaneous malignant melanoma with lipoblast-like cells. PMID- 12653590 TI - Pathologic quiz case: a 4-year-old girl with multiple skin nodules. PMID- 12653589 TI - Gamna-Gandy nodules in a renal clear cell carcinoma. PMID- 12653591 TI - Pathologic quiz case: post-lung transplantation patient with bilateral breast masses. PMID- 12653592 TI - Pathologic quiz case: a solitary pulmonary nodule in a young woman. PMID- 12653593 TI - Pathologic quiz case: a 77-year-old man with incidental discovery of pancreatic mass on abdominal computed tomography. PMID- 12653596 TI - Anaplastic large cell lymphoma arising in a silicone breast implant capsule: a case report and review of the literature. AB - Anaplastic large cell lymphoma is a rare type of primary breast lymphoma. We report a case of anaplastic large cell lymphoma, T-cell phenotype, occurring in the periprosthetic capsule of a silicone breast prosthesis 9 years after implantation for augmentation mammoplasty. This case is unique for its unusual presentation. PMID- 12653598 TI - Metastatic renal cell carcinoma to the ovary: a case report and discussion of differential diagnoses. AB - The clinical presentation of renal cell carcinoma in the ovary is rare and may closely resemble that of other ovarian clear cell tumors. Metastatic renal cell carcinoma was diagnosed in a patient who presented with a right adnexal mass and no other significant past medical history. The gross appearance of the tumor suggested a primary ovarian neoplasm. However, microscopic and immunohistochemical analysis revealed involvement of both ovaries by a clear cell tumor with features of renal cell carcinoma. A renal mass was subsequently discovered. We discuss the clinical, histologic, and immunohistochemical features of this tumor and the features of other clear cell neoplasms in the differential diagnosis. PMID- 12653597 TI - Hepatosplenic alpha beta T-cell lymphoma: a report of an S100-positive case. AB - Hepatosplenic T-cell lymphoma is an uncommon neoplasm characterized by a lymphoid infiltrate within the sinusoids of the liver, spleen, and bone marrow, without significant nodal involvement. The majority of cases express the gammadelta T cell receptor and are associated with an isochromosome 7q cytogenetic abnormality. Recently, a small number of cases have been reported that express the alphabeta T-cell receptor. Here, we report our findings of a case of an S100 positive hepatosplenic alphabeta T-cell lymphoma in a 20-year-old woman who presented with pancytopenia and hepatosplenomegaly. The case adds to the growing literature of hepatosplenic alphabeta T-cell lymphomas. PMID- 12653599 TI - Inflammatory pseudotumor of the spleen in a 6-year-old child: a clinicopathologic study. AB - Inflammatory pseudotumors of the spleen are extremely rare in children. To our knowledge, only 3 cases of splenic inflammatory pseudotumors have been reported. An inflammatory pseudotumor of the spleen was found incidentally during the workup of vesicoureteral reflux disease in a 6-year-old girl. The clinical, pathologic, immunophenotypic, and molecular features and the differential diagnostic features are discussed here. Splenic inflammatory pseudotumors, although rare, should be considered in the differential diagnosis of a mass lesion of the spleen in children. PMID- 12653600 TI - Diffuse neuroendocrine differentiation in a morphologically composite mammary infiltrating ductal carcinoma: a case report and review of the literature. AB - Neuroendocrine differentiation has been reported in both in situ and infiltrating breast cancers. The prognostic significance of neuroendocrine differentiation in mammary carcinoma is unclear. We report a case of infiltrating ductal carcinoma in which there was a morphologically conventional-appearing infiltrating ductal component admixed with nests of cells that resembled a carcinoid tumor and initially mimicked the appearance of intraductal carcinoma. Immunohistochemical stains for synaptophysin and chromogranin demonstrated diffuse, strong positivity uniformly throughout the tumor, even in the more conventional-appearing areas. Electron microscopic examination of tissue retrieved from paraffin blocks was attempted unsuccessfully. We concluded that this was an infiltrating ductal carcinoma with morphologic and immunohistochemical evidence of neuroendocrine differentiation. The case is discussed with a review of the literature and a discussion of nomenclature for tumors of the breast showing variable degrees of neuroendocrine differentiation. PMID- 12653601 TI - Renal medullary carcinoma: a report of 2 cases and review of the literature. AB - Renal medullary carcinoma is a recently described aggressive neoplasm of the kidney. With the exception of 2 patients, all other reported cases have been associated with sickle cell hemoglobinopathies, mainly sickle cell trait and hemoglobin SC disease. Renal medullary carcinoma is a highly malignant tumor with evidence of angiolymphatic and distant metastasis at the time of diagnosis. No specific genetic abnormality has been identified in this neoplasm despite its close association with a genetic disease. We describe 2 cases of renal medullary carcinoma, one associated with hemoglobin SC disease, and the other with what we believe to be the first reported case associated with sickle cell disease. PMID- 12653602 TI - Duodenal gangliocytic paraganglioma with lymph node metastasis: a case report and review of the literature. AB - A case of duodenal gangliocytic paraganglioma (DGP) in a 67-year-old woman is presented. The DGP arose in the second part of the duodenum. Although most of the reported cases of DGP are considered benign, in the present case, we found regional lymph nodes containing metastatic tumor. Previous reports have documented metastases containing only epithelioid cells. The current case demonstrates metastatic tumor in regional lymph nodes containing all 3 of the DGP components (spindle cells, ganglion-like cells, and epithelioid cells). PMID- 12653603 TI - Natural killer-like T-cell lymphoma of the small intestine with a distinct immunophenotype and lack of association with gluten-sensitive enteropathy. AB - We report the case of a large natural killer (NK)-like T-cell lymphoma that involved the ileum and displayed a distinct immunophenotype and complex karyotype. The patient exhibited no evidence of gluten-sensitive enteropathy (celiac disease) or any other type of enteropathy as determined by clinical history, endoscopy, and serology for immunoglobulin A (IgA) antiendomysial and IgG antigliadin antibodies. Molecular studies demonstrated a clonal T-cell receptor gamma chain gene rearrangement. Immunophenotype analysis showed expression of intestinal epithelium-homing receptor CD103, CD7, cytoplasmic CD3 epsilon, CD56, and CD16 but no other T- or NK-cell markers. Cytogenetic analysis of the malignant cells revealed multiple chromosomal abnormalities indicative of a biologically advanced, high-grade lymphoma. A novel subset of normal intestinal intraepithelial lymphocytes, bearing a similar phenotype, has been described; moreover, this subset diminishes, rather than expands, in gluten-sensitive enteropathy. This case supports the notion that lymphomas involving the small intestine represent a heterogeneous group of lymphomas with diverse pathogenetic mechanisms. PMID- 12653604 TI - Malignant peripheral nerve sheath tumor with divergent differentiation. AB - Right posterior thigh malignant peripheral nerve sheath tumor with malignant glandular and rhabdomyoblastic components was diagnosed in a 24-year-old African American man. Malignant glands and rhabdomyoblastic cells were admixed with the spindle cells. Immunohistochemistry demonstrated spindle cells positive for S100 and vimentin; the glandular component was positive for chromogranin, and rare cells were positive for CK20 but negative for CK7. Rhabdomyoblastic cells were positive for muscle-specific actin and desmin. A single pulmonary metastasis occurred 5 months after surgical resection of the tumor. Death occurred 1 month later and was probably due to a pulmonary embolus. PMID- 12653605 TI - Secondary tumors in mammary adenolipomas: a report of 2 unusual cases. AB - Two unusual examples of secondary tumors arising in adenolipomas were diagnosed in a 16-year-old girl and a 27-year-old woman, each of whom presented with asymmetric enlargement of the right breast. In both instances, a significant secondary lesion was associated with the adenolipoma. In the first case, a cyst appeared to have arisen in a fibroadenoma within the adenolipoma, which had apparently undergone infarction during a recent pregnancy and postpartum period. There was micropapillary duct epithelial hyperplasia of the cyst lining epithelium. In the second case, an adenofibroma with atypical lobular hyperplasia was present within the adenolipoma. PMID- 12653606 TI - Metastatic infiltrating syringomatous adenoma of the breast. AB - Infiltrating syringomatous adenoma of the breast is classified as a benign, locally invasive neoplasm. To our knowledge, there have been no reported cases of metastasizing syringomatous adenomas. We used sentinel lymph node mapping to identify metastatic cells, leading to a diagnosis of infiltrating syringomatous adenoma of the breast. PMID- 12653607 TI - Infection by Dipylidium caninum in an infant. AB - Dipylidium caninum, the dog tapeworm, is a cosmopolitan parasite of dogs and cats and occasionally causes human infection in the United States. Diagnosis is made by observing the characteristic rice grain-like proglottids in stool specimens and the pathognomonic egg packets in the gravid uterus in histologic sections of the parasite. There have been few reported cases of human infection with this parasite, and very little information on the pathology of this zoonotic disease is available in the English language. This report of a case of D caninum infection in a 6-month-old infant highlights the diagnostic features of this disease. To our knowledge, this is the first case to be reported in the American pathology literature during the last 36 years (MEDLINE database, 1966-2002). PMID- 12653608 TI - Pathologic quiz case: a pulmonary mass. PMID- 12653609 TI - Pathologic quiz case: a 16-year-old girl with a palpable liver mass, fever, and vomiting. PMID- 12653610 TI - Pathologic quiz case: a 24-year-old man with cutaneous and mandibular lesions. PMID- 12653611 TI - Pathologic quiz case: a 55-year-old woman with an adrenal mass. PMID- 12653612 TI - Pathologic quiz case: a 23-year-old woman with an ovarian mass. PMID- 12653613 TI - Pathologic quiz case: right thigh pain for 14 months. PMID- 12653614 TI - Pathologic quiz case: a 28-year-old man with an impacted tooth. PMID- 12653615 TI - Pathologic quiz case: a large pelvic mass in a 44-year-old man. PMID- 12653616 TI - The health effects of soft drinks. PMID- 12653617 TI - Promoting culturally competent care for gay youth. AB - Gay youth and those questioning their sexual identity have been referred to as "hidden," "invisible," "stigmatized," and "marginalized." As a result, the unique safety and health needs of this subculture have been overlooked, or worse, ignored, placing these youth at risk. Because school nurses have been identifying at-risk populations of students and developing programs to promote youth and family health for years, they should be prepared to provide health care for the subculture of gay youth. However, nurses are saying they do not have the knowledge or skills needed to identify and address the needs of this group. Providing school nursing care for gay youth requires the school nurse to be culturally competent. School nurses need to be aware of, sensitive to, and knowledgeable about the subculture. They must also possess communication skills required to relate appropriately to this group. This article presents information and nursing strategies that will promote the safety and health of gay youth while enhancing the school nurse's cultural competence. PMID- 12653618 TI - The health of the school nurse community: a framework. AB - School nursing is based on a conceptual foundation of community health nursing. Using community health nursing as a reference point, this article describes a viewpoint of school nurses as the population of care. With this perspective, school nurses will better understand how to foster the health of their community. Developed on the basis of McKenzie, Pinger, and Kotecki's community health model, the Health of the School Nurse Community Framework has been designed to (a) facilitate understanding of the concept of "school nurse community" and (b) organize the factors that affect the health of the school nurse community. Essential to its use is the identification of the school nurse community's assets and capacities as well as its needs. Application of the framework to strive to improve the health of one state's school nurse community is also presented. PMID- 12653619 TI - A Lyme disease case study and individualized healthcare plan. AB - The Atlantic and Pacific coasts are the boundaries of Lyme disease with the Northeastern and Midwestern regions of the United States continuing to report the majority of cases. New reported cases of Lyme disease doubled from 1991 to 2001 according to statistics published by the Centers for Disease Control and Prevention. Within that population are more children between the ages of 5 and 9. The younger the child, the more difficult it is to diagnose Lyme disease. Children under the age of 19 are at high risk for Lyme disease because of the amount of time spent in outdoor activities during the late spring and summer. These months correlate with the tick breeding cycle and the peak time for outdoor recreational activities. Lyme disease can pose serious health risks in late stage illness. A Lyme disease case study on a school-age child provides comprehensive assessment data, interventions, and educational information for parents. A template of an individualized healthcare plan using standardized language is a guide for school nurses. The value of school nurses as guardians of the public's health is addressed. PMID- 12653620 TI - Development of a skin cancer prevention program. AB - The Centers for Disease Control and Prevention (CDC) now categorizes skin cancer as epidemic. Nearly 90% of these deadly cancers start from sun exposure during the childhood years. This makes sun exposure in school-age children a serious public health risk, also one that school nurses can address. Solar radiation is now classified as a "known human carcinogen," with ultraviolet rays joining the ranks of other known cancer-causing elements such as tobacco, arsenic, and radon. Sun exposure to unprotected skin results in painful burns, premature aging, cataracts, and weakened immune systems. School nurses can use the new CDC guidelines along with other local and state resources to develop a specific skin cancer prevention program for their schools. They are in a pivotal position to partner with students, parents, administrators, teachers, and coaches to reduce the occurrence of skin cancer in children. The article describes one high school's skin cancer prevention project. PMID- 12653621 TI - Evaluation of the SunWise School Program. AB - Melanoma, the most fatal form of skin cancer, is rising at rates faster than all other preventable cancers in the United States. Childhood exposure to ultraviolet (UV) light increases the risk for skin cancer as an adult, therefore, starting positive sun protection habits early may be key to reducing the incidence of this disease. The Environmental Protection Agency's SunWise School Program, a national environmental and health education program for sun safety of children in primary and secondary schools (Grades K-8), was evaluated. Pretests (n = 4,559) and posttests (n = 4,016) were completed by students from 85 schools in 35 states. Significant increases were noted for the three knowledge variables. Intentions to play in the shade increased from 70% to 76% (p < 0.001) with more modest changes in intentions to use sunscreen. Attitudes regarding the healthiness of a tan also decreased significantly. Brief, standardized sun protection education can be efficiently interwoven into existing school curricula and result in increases in knowledge and positive intentions for sun protection. PMID- 12653622 TI - A bicycle safety education program for parents of young children. AB - This study examined parental perceptions of the benefits and barriers to bicycle helmet use and their level of knowledge about bicycle safety issues. A school based bicycle safety education program was taught to first- and second-grade students in a rural/suburban school district by a graduate nursing student. Pender's Health Promotion Model was the theoretical framework used to guide this research study. A parent Bicycle Helmet Questionnaire (BHQ) developed by the author was the tool used to gather data. The study determined that parents of children in schools that received the educational intervention prior to completing the questionnaire had significantly higher knowledge level scores and that the majority of parents were in favor of bicycle helmet legislation for children. The implications from this research study include the importance of bicycle safety education interventions for students and parents and the need to implement bicycle helmet legislation. PMID- 12653623 TI - Vision screening practices in central Iowa: a follow-up evaluation. AB - In response to data from the initial survey published in October 2001, interventions were developed to increase vision screening knowledge and promote use of the Iowa Vision Screening Program Guidelines. The purpose of this evaluation was to measure differences in practice after interventions, including attendance at a half-day workshop and practice session. Results of the study indicate that participants of the workshop and practice session were more likely to conduct yearly distance and near vision, and depth perception screening. In addition, they were more likely to use the recommended screening instruments for distance and near vision, and depth perception. They were also much more likely to use recommended referral criteria for all types of screening and to have changed some aspect of their vision screening practice in the past year. Results are presented with implications for school nursing practice. PMID- 12653625 TI - Genes in food--why the furore? AB - Although unprecedented and perhaps unique in its irrationality, the recent furore over genetically modified (GM) food holds extremely important lessons for scientists. Some sections of the media undoubtedly bear a heavy responsibility for giving the expression 'GM' threatening connotations that are quite unwarranted. However, influential contributions to the hysteria have come from a surprising range of other sources, including some scientists. The research community has failed in its responsibility to society in three ways. Firstly, plant scientists did not appreciate that certain techniques (such as the use of antibiotic resistance genes as markers during plant transformation) would inevitably provoke public consternation. As a result, they took no steps to address such concerns. Secondly, researchers overlooked, minimized or in some cases simply dismissed the significance of public fears that they were 'interfering with Nature' or 'playing God'. Thirdly, plant breeders apparently saw no need to take pro-active measures with regard to the media and public in placing potential environmental and nutritional benefits of GM crops on the agenda in a positive fashion. Partly because of this failure, GM food is now firmly established in the public mind as wholly objectionable. One measure of how far we have travelled down that road is that it hardly matters any more whether objections are based on alleged environmental risks of cultivating GM crops or alleged toxicological hazards of eating them. 'Genetically modified organism', like 'radioactivity', has become an odious, generic shibboleth. Given that millions of people throughout the world are already benefiting from pharmaceuticals made by GM organisms, this is bizarre. PMID- 12653626 TI - How to (or not to) communicate science. AB - Protagonists for 'the public understanding of science' still sometimes fail to recognize that there is also a need for 'the scientists' understanding of the public' and that for most of science most of the time we are all public. 'Science' is communicated to 'the public' through popular books, museums, TV, the Internet, but far too often the present state of scientific belief is presented uncritically as the onward march of truth as discovered by Euro-American males. This has contributed to a widespread public concern, if not mistrust, in many areas of science, not least genetics and neuroscience. Although researchers often criticize the media for misrepresenting their work, the hype and simplifications often begin with the press releases put out by the researchers, their institutions and the scientific journals themselves. I conclude by looking more optimistically at the ways in which, by bringing natural science into theatre, novels and other art forms, the fragmentation of our culture may be diminished. PMID- 12653627 TI - Sulphated endothelial ligands for L-selectin in lymphocyte homing and inflammation. AB - Lymphocytes from the blood home to secondary lymphoid tissues through a process of tethering, rolling, firm adhesion and transmigration. Tethering and rolling of lymphocytes is mediated by the interaction of L-selectin on lymphocytes with sulphated ligands expressed by the specialized endothelial cells of high endothelial venules (HEVs). The sulphate-dependent monoclonal antibody MECA79 stains HEVs in peripheral lymph nodes and recognizes the complex of HEV ligands for L-selectin termed peripheral node addressin. High endothelial cell GlcNAc-6 sulphotransferase/L-selectin ligand sulphotransferase is a HEV-expressed sulphotransferase that contributes to the formation of the MECA79 epitope and L selectin ligands on lymph node HEVs. MECA79-reactive vessels are also common at sites of chronic inflammation, suggesting mechanistic parallels between lymphocyte homing and inflammatory trafficking. PMID- 12653628 TI - Sulphotransferases acting on mucin-type oligosaccharides. AB - This review summarizes the occurrence, properties and role in mucin O glycosylation pathways of the various members of glycoprotein sulphotransferase families. Although a number of sulphotransferases have been cloned that act on mucin-type substrates in vitro, it is still difficult to determine exactly which enzymes are responsible for mucin sulphation in vivo. Sulphotransferases play a critical role in determining the chemical, physical and biological properties of mucins. Several of these enzymes have been shown to differ in expression and activity in cancer and inflammation. PMID- 12653629 TI - Glycoprotein hormone GalNAc-4-sulphotransferase. AB - The glycoprotein hormones lutropin (LH) and thyrotropin and a limited number of additional glycoproteins bear carbohydrate structures terminating with the unique sequence SO(4)-4-GalNAcbeta1,4GlcNAcbeta that has been conserved in the glycoprotein hormones of all vertebrate species. Synthesis of these structures is mediated by a protein-specific beta1,4GalNAc-transferase and a GalNAc-4 sulphotransferase (GalNAc-4-ST1). GalNAc-4-ST1 is a member of a family of sulphotransferases that includes HNK-1 sulphotransferase, chondroitin-4 sulphotransferases-1-3 and dermatan-4-sulphotransferase-1. With the exception of HNK-1-ST, these sulphotransferases add sulphate to the C-4 hydroxy group of either terminal or non-terminal beta1,4-linked GalNAc. GalNAc-4-ST1 is most highly expressed in pituitary, cerebellum and other regions of the brain. The terminal GalNAcSO(4) on LH is recognized by the cysteine-rich domain of the mannose/GalNAc-4-SO(4) receptor located in hepatic endothelial cells. Each cysteine-rich domain binds a single terminal GalNAc-4-SO(4), and the receptor must form non-covalently associated homodimers in order to simultaneously engage two GalNAc-4-SO(4) moieties on separate oligosaccharides with sufficient affinity to form stable complexes. The receptor mediates the clearance of LH from the blood. This clearance, in conjunction with the stimulated release of hormone from dense core granules in pituitary gonadotroph cells, is required to produce the episodic rise and fall in LH levels needed for optimal oestrogen production during the implantation of embryos in the uterus. PMID- 12653630 TI - Heparan sulphate N-sulphotransferase activity: reaction mechanism and substrate recognition. AB - Human heparan sulphate N-deacetylase/N-sulphotransferase 1 sulphates the NH(3) (+) group of the glucosamine moiety of the heparan chain in heparan sulphate/heparin biosynthesis. An open cleft that runs perpendicular to the sulphate donor 3'-phosphoadenosine 5'-phosphosulphate may constitute the acceptor substrate-binding site of the sulphotransferase domain (hNST1) [Kakuta, Sueyoshi, Negishi and Pedersen (1999) J. Biol. Chem. 274, 10673-10676]. When a hexasaccharide model chain is docked into the active site, only a trisaccharide ( IdoA-GlcN-IdoA-) portion interacts directly with the cleft residues: Trp-713, His 716 and His-720 from alpha helix 6, and Phe-640, Glu-641, Glu-642, Gln-644 and Asn-647 from random coil (residues 640-647). Mutation of these residues either abolishes or greatly reduces hNST1 activity. Glu-642 may play the critical role of catalytic base in the sulphuryl group transfer reaction, as indicated by its hydrogen-bonding distance to the NH(3) (+) group of the glucosamine moiety in the model and by mutational data. PMID- 12653631 TI - Chondroitin sulphate proteoglycans in the central nervous system: changes and synthesis after injury. AB - Chondroitin sulphate proteoglycans (CSPGs) are up-regulated in the central nervous system after injury, specifically around the lesion site where the glial scar forms. This structure contains astrocytes, oligodendrocyte precursor cells, microglia and meningeal cells, and forms an inhibitory substrate for axon re growth. CSPGs have been shown to be closely involved in this neuronal growth inhibition, specifically through their sugar chains. These chains are composed of repeats of the same disaccharide unit carrying sulphate groups in different positions. The sulphation pattern directly influences the CSPG binding properties and function; the specific sulphation pattern required for the inhibitory activity of these molecules on axon growth is unknown at present. The expression of the chondroitin sulphotransferases, which sulphate the disaccharide residues of CSPGs and thus are responsible for the structural diversity of the chondroitin sulphate sugar chains, is regulated differently in central nervous system during development and after injury, suggesting the implication of a specific sulphation pattern in the inhibitory activity of CSPGs. PMID- 12653632 TI - Interaction between glial-cell-line-derived neurotrophic factor (GDNF) and 2-O sulphated heparin-related glycosaminoglycans. AB - Glial cell line-derived growth factor (GDNF) is a cytokine of the transforming growth factor (TGF)-beta family with potent neuroprotective activity. Clinical trials of recombinant GDNF in advanced Parkinson's disease are currently under way. It is known that mice homozygous for disruption of the gene encoding heparan sulphate 2-O-sulphotransferase die perinatally, due to the complete absence of kidneys. Similar phenotypes arise from targeted disruption of the genes encoding either GDNF, or its receptors, GFRalpha1 and c-Ret. It may therefore be proposed that GDNF normally binds to 2-O-sulphate-rich heparan sulphate within kidney progenitor tissues, and that this interaction is essential for its activity in kidney development. In support of this hypothesis we have shown in ELISA studies that GNDF binds to heparin and heparan sulphate. This binding is unusually sensitive to the chemical 2-O-desulphation, and promotes the binding of GNDF to GFRalpha1. PMID- 12653634 TI - Heparan sulphate sulphotransferase expression in mice and Caenorhabditis elegans. AB - Heparan sulphate (HS) acts as a multifunctional cell regulator, with specific sulphated saccharide sequences designed for selective interactions with many proteins. Functionally, these interactions result in regulation of the protein activities, and there is growing evidence that cells can dynamically alter the structure of HS sequences that they display. HS biosynthesis involves the action of a complex set of enzymes with polymerase, epimerase and sulphotransferase (ST) activities. In higher organisms, multiple isoforms of STs decorate the nascent HS chains with specific patterns of sulphation that confer selective biological functions. The study of HSSTs in model organisms provides a valuable opportunity to examine the expression of these enzymes in relation to the structure and activities of the HS produced. Here we describe that, in mice, there are stage specific combinations of HSST isoenzymes that underlie the synthesis of different HS species at different times in the developing brain. This differential expression of HSSTs results in the synthesis of structurally variant HS species that form functional signalling complexes with specific fibroblast growth factors and their receptors. Regulated synthesis of specific HS species could be a mechanism for the regulation of proliferation and differentiation in the developing brain. We also describe evidence that a Caenorhabditis elegans orthologue of the mammalian 2OST enzyme, called HST-2, is essential for the normal development of this nematode. Together, these studies emphasize the importance of HSSTs in the biosynthesis of functionally variant HS proteoglycans, and demonstrate the importance of these complex regulatory molecules in developmental processes. PMID- 12653633 TI - Glucosaminyl N-deacetylase/N-sulphotransferases in heparan sulphate biosynthesis and biology. AB - During the biosynthesis of heparan sulphate (HS) in the Golgi compartment, the first modification enzyme, glucosaminyl N-deacetylase/N-sulphotransferase (NDST), starts to work on the growing HS polysaccharide chain. This enzyme defines the overall design of the sulphation pattern, which will determine the ability of the HS chain to interact with target molecules. NDST removes acetyl groups from glucosamine residues and replaces them with sulphate groups. These N-sulphate groups are essential for further modification during biosynthesis; without N sulphation, no O-sulphation or conversion of glucuronic acid into iduronic acid will occur. Four NDST isoforms, transcribed from four genes, have been identified. Much of our work is concentrated on how the enzymes are organized within the Golgi compartment and the identification of interacting partners. In addition, we study mice in which the gene encoding NDST-1 or NDST-2 has been knocked out. NDST-1 knockout mice with altered HS structure die at birth due to lung failure, whereas lack of NDST-2 results in abnormal mast cells. Since NDSTs have a key role in HS design (see above), these mice can be used to study HS function. Areas of interest are cell differentiation, growth, inflammation, cancer, lipid metabolism and microbial infection. PMID- 12653635 TI - A gravimetric analysis of protein-oligosaccharide interactions. AB - Interactions between an immobilized, heparin-derived octasaccharide and growth factors have been observed using a quartz crystal microbalance-dissipation (QCM D). This device can measure the amount of growth factors binding to the octasaccharide surface and also the change of dissipation of the surface. Dissipation is a measure of how the adhered material 'damps' the surface vibrations. The octasaccharides were anchored through their reducing ends by the intermediary of the alkanethiol molecule, which covalently binds to the crystal surface through the thiol group. As expected, heparin sulphate binding growth factors bound to the octasaccharide, but the change in mass of growth factor bound per unit change in dissipation is different for the different growth factors. Suggesting that the structures of the various growth factor octasaccharide complexes are different, therefore, indicates that the change in dissipation can give insights into the structure, orientation and packing of the oligosaccharide-growth factor complexes. PMID- 12653636 TI - Hepatocyte growth factor/scatter factor and its interaction with heparan sulphate and dermatan sulphate. AB - Hepatocyte growth factor (HGF)/scatter factor (SF) is a unique growth factor, in that it binds both heparan sulphate (HS) and dermatan sulphate (DS). The sequences in HS and DS that specifically interact with and modulate HGF/SF activity have not yet been fully identified. Ascidian DS, which uniquely possesses O-sulphation at C-6 (and not C-4) of its N -acetylgalactosamine unit, was analysed for HGF/SF-binding activity in the biosensor. The kinetic analysis revealed a strong, biologically relevant interaction with an equilibrium dissociation constant ( K (d)) of approx. 1 nM. An Erk activation assay also demonstrated stimulation of the MAP kinase pathway downstream of the Met receptor following addition of both HGF/SF and ascidian DS to the glycosaminoglycan deficient CHO-745 mutant cell line. Furthermore, the activation of Met and the MAP kinase pathway by HGF/SF and ascidian DS leads to a cellular response in the form of migration. PMID- 12653637 TI - Polyamines and their role in human disease--an introduction. AB - The naturally occurring polyamines are found in all living cells, where they fulfil a number of critical functions in relation to cell growth. The quest to identify these functions has been the subject of five independent colloquia hosted by the Biochemical Society and today still occupies several hundred scientists across Europe, the U.S.A. and Japan. PMID- 12653638 TI - Transgenic mouse models for studies of the role of polyamines in normal, hypertrophic and neoplastic growth. AB - Transgenic mice expressing proteins altering polyamine levels in a tissue specific manner have considerable promise for evaluation of the roles of polyamines in normal, hypertrophic and neoplastic growth. This short review summarizes the available transgenic models. Mice with large increases in ornithine decarboxylase (ODC), S-adenosylmethionine decarboxylase or antizyme, a protein regulating polyamine synthesis by reducing polyamine transport and ODC in the heart, have been produced using constructs in which the protein is expressed from the alpha -myosin heavy-chain promoter. These mice are useful in studies of the role of polyamines in hypertrophic growth. Expression from keratin promoters has been used to target increased synthesis of ODC, spermidine/spermine-N(1) acetyltransferase (SSAT) and antizyme in the skin. Such expression of ODC leads to an increased sensitivity to chemical and UV carcinogenesis. Expression of antizyme inhibits carcinogenesis in skin and forestomach. Expression of SSAT increases the incidence of skin papillomas and their progression to carcinomas in response to a two-stage carcinogenesis protocol. These results establish the importance of polyamines in carcinogenesis and neoplastic growth and these transgenic mice will be valuable experimental tools to evaluate the importance of polyamines in mediating responses to oncogenes and studies of cancer chemoprevention. PMID- 12653640 TI - Polyamine dependence of normal cell-cycle progression. AB - The driving force of the cell cycle is the activities of cyclin-dependent kinases (CDKs). Key steps in the regulation of the cell cycle therefore must impinge upon the activities of the CDKs. CDKs exert their functions when bound to cyclins that are expressed cyclically during the cell cycle. Polyamine biosynthesis varies bicyclically during the cell cycle with peaks in enzyme activities at the G(1)/S and S/G(2) transitions. The enzyme activities are regulated at transcriptional, translational and post-translational levels. When cells are seeded in the presence of drugs that interfere with polyamine biosynthesis, cell cycle progression is affected within one cell cycle after seeding. The cell cycle phase that is most sensitive to polyamine biosynthesis inhibition is the S phase, while effects on the G(1) and G(2)/M phases occur at later time points. The elongation step of DNA replication is negatively affected when polyamine pools are not allowed to increase normally during cell proliferation. Cyclin A is expressed during the S phase and cyclin A/CDK2 is important for a normal rate of DNA elongation. Cyclin A expression is lowered in cells treated with polyamine biosynthesis inhibitors. Thus, polyamines may affect S phase progression by participating in the regulation of cyclin A expression. PMID- 12653639 TI - The role of polyamine catabolism in anti-tumour drug response. AB - Interest in polyamine catabolism has increased since it has been directly associated with the cytotoxic response of multiple tumour types to exposure to specific anti-tumour polyamine analogues. Human polyamine catabolism was considered to be a two-step pathway regulated by the rate-limiting enzyme spermidine/spermine N(1)-acetyltransferase (SSAT) that provides substrate for an acetylpolyamine oxidase (APAO). Further, the super-induction of SSAT by several anti-tumour polyamine analogues has been implicated in the cytotoxic response of specific solid-tumour phenotypes to these agents. This high induction of SSAT has been correlated with cellular response to the anti-tumour polyamine analogues in several systems and considerable progress has been made in understanding the molecular mechanisms that regulate the analogue-induced expression of SSAT. A polyamine response element has been identified and the transacting transcription factors that bind and stimulate transcription of SSAT have been cloned and characterized. The link between SSAT activity and cellular toxicity is thought to be based on the production of H(2)O(2) by the activity of the constitutive APAO that uses the SSAT-produced acetylated polyamines. The high induction of SSAT and the subsequent activity of APAO are linked to the cytotoxic response of some tumour cell types to specific polyamine analogues. However, we have recently cloned a variably spliced human polyamine oxidase (PAOh1) that is inducible by specific polyamine analogues, efficiently uses unacetylated spermine as a substrate, and also produces toxic H(2)O(2) as a product. The results of studies with PAOh1 suggest that it is an additional enzyme in polyamine catabolism that has the potential to significantly contribute to polyamine homoeostasis and drug response. Most importantly, PAOh1 is induced by specific polyamine analogues in a tumour-phenotype-specific manner in cell lines representative of the major forms of solid tumours, including lung, breast, colon and prostate. The sensitivity to these anti-tumour polyamine analogues can be significantly reduced if the tumour cells are co-treated with 250 microM of the polyamine oxidase inhibitor N (1), N (4)-bis(2,3-butadienyl)-1,4-butanediamine (MDL 72,527), suggesting that the H(2)O(2) produced by PAOh1 does in fact play a direct role in the observed cytotoxicity. These results strongly implicate PAOh1 as a new target that, in combination with SSAT, may be exploited for therapeutic advantage. The current understanding of the role and regulation of these two important polyamine catabolic enzymes are discussed. PMID- 12653641 TI - Acrolein produced from polyamines as one of the uraemic toxins. AB - It is well known that the addition of spermine or spermidine to culture medium containing ruminant serum inhibits cellular proliferation. This effect is caused by the products of oxidation of polyamines that are generated by serum amine oxidase. Among the products, we found that acrolein is a major toxic compound produced from spermine and spermidine by amine oxidase. We then analysed the level of polyamines (putrescine, spermidine and spermine) and amine oxidase activity in plasma of patients with chronic renal failure. It was found that the levels of putrescine and the amine oxidase activity were increased, whereas spermidine and spermine were decreased in plasma of patients with chronic renal failure. The levels of free and protein-conjugated acrolein were also increased in plasma of patients with chronic renal failure. An increase in putrescine, amine oxidase and acrolein in plasma was observed in all cases such as diabetic nephropathy, chronic glomerulonephritis and nephrosclerosis. These results suggest that acrolein is produced during the early stage of nephritis through kidney damage and also during uraemia through accumulation of polyamines in blood due to the decrease in their excretion into urine. PMID- 12653642 TI - Polyamines and prostatic cancer. AB - The importance of polyamines in prostatic growth and differentiation has prompted studies to evaluate the clinical relevance of the ornithine decarboxylase/polyamine system in prostatic cancer. These studies show that differences in biological behaviour of prostatic (cancer) cells are associated with changes in polyamine levels and/or the activity of their metabolic enzymes. Faulty antizyme regulation of polyamine homoeostasis may play an important role in the growth and progression of prostatic carcinoma. Treatment of human prostate carcinoma cells with inhibitors of polyamine metabolic enzymes or polyamine analogues induces cell growth arrest or (apoptotic) cell death. Our recent in vitro studies using conformationally restricted polyamine analogues show that these compounds inhibit cell growth, probably by inducing antizyme-mediated degradation of ornithine decarboxylase. Sensitivity of human prostate cancer cells for these compounds was increased in the absence of androgens. These results suggest that these analogues might have chemotherapeutic potential in case prostatic cancer has become androgen-independent. Pilot data in an in vivo model show that these analogues have effects on tumour cell proliferation, vascularity, blood perfusion and tissue hypoxia. Overall, these studies show that polyamines may serve as important biomarkers of prostatic malignancy and provide a promising target for chemotherapy of prostatic cancer. PMID- 12653643 TI - Polyamines and colon cancer. AB - In colon cancer, the activities of polyamine-synthesizing enzymes and polyamine content are increased 3-4-fold over that found in the equivalent normal colonic mucosa, and polyamines have even been attributed as markers of neoplastic proliferation in the colon. Furthermore, and in contrast with all other cell systems in the body, normal and neoplastic cells in the colon are exposed to high concentrations of putrescine from the lumen, synthesized by colonic microflora. While such a high polyamine supply may be of benefit in non-neoplastic colonic mucosal growth, the role of luminal polyamines in colon cancer is a clear concern. Luminal polyamines are readily taken up by neoplastic colonocytes, they are utilized in full to support neoplastic growth, and their uptake is strongly up-regulated by the mitogens known to play an important role in colonic carcinogenesis. Inhibition of polyamine synthesis and their uptake, impaired utilization of exogenous polyamines, and enhanced catabolism of polyamines in neoplastic colonocytes are therefore logical approaches in the chemoprevention of colorectal cancer. PMID- 12653644 TI - Polyamine-reduced diet in metastatic hormone-refractory prostate cancer (HRPC) patients. AB - Polyamine (PA) deprivation is effective in prostate carcinoma models. We have assessed the observance by patients, tolerance and side effects of a PA-reduced diet (PRD) and intestinal decontamination (ID), in order to reduce PA dietary and intestinal bacterial pools, in metastatic, hormone-refractory prostate cancer (HRPC) patients. A total of 13 volunteers (mean age, 67+/-10 years) with metastatic HRPC were proposed for PRD and ID (0.75 g/day of oral neomycin every other week). The mean time from HRPC diagnosis to the start of the diet was 12+/ 8 months. Of the total 13, seven patients had received prior chemotherapy or Estramustine phosphate. PRD was obtained after HPLC assessment of PA contents in current foods and given 5 days a week. Toxicity, performance and pain status were assessed according to the World Health Organisation and EORTC scales. Prostatic specific antigen (PSA), blood counts, ionograms, transaminases and erythrocyte PA spermidine (Spd) and spermine (Spm; assessed by HPLC) were evaluated regularly. Mean observance was 8+/-7 months (range, 2-26 months). One case of grade II toxicity to neomycin was observed. Cancer-specific survival (after the diet) was 14+/-7 months, and two patients are still alive. All the other patients have died of their cancer at 12+/-6 months (range, 4-20 months). Cancer-specific survival after hormonal escape was 27+/-11 months (range, 9-45 months). Performance status was improved during the regimen and deteriorated 3 months after stopping. Pain score was improved (1.3 versus 0.6; P =0.04) during the diet and increased (2.1 versus 0.3) 3 months after stopping. Erythrocyte Spd (11.6+/-7 versus 7.7+/-2 nmol/8 x 10(9) erythrocytes; P =0.036) and Spm (7+/-6 versus 3.9+/-1.6 nmol/8 x 10(9) erythrocytes; P =0.036) levels were significantly reduced at 3 months. One patient had a >50% reduction in PSA, three patients had PSA stabilization for 6 months. PSA progression was observed in all other patients. No significant modification of other studied biological parameters was noted. Reducing PA dietary intake and ID is a well-observed and tolerated regimen and seems to be beneficial for patient quality of life and pain control. Patients with low initial PSA can experience durable stabilization. These encouraging results in such an aggressive disease certainly warrant further investigation. PMID- 12653645 TI - Polyamines as modifiers of genetic risk factors in human intestinal cancers. AB - Polyamines are downstream mediators of genetic risk factors in human intestinal cancers. The adenomatous polyposis coli (APC) tumour-suppressor gene, which is mutated in essentially all human colon cancers, regulates the expression of several e-box transcription factors. These factors, in turn, regulate the transcription of ornithine decarboxylase (ODC), the first enzyme in polyamine synthesis. The Kirsten ras ( K-ras ) oncogene regulates the expression of several genes, including suppressing the expression of peroxisomal proliferator-activated receptor gamma (PPARgamma). This PPAR, in turn, activates the expression of the spermidine/spermine-N(1)-acetyltransferase (SSAT), the first enzyme in polyamine catabolism. The non-steroidal anti-inflammatory drug (NSAID) sulindac induces the transcription of SSAT via activation of PPARgamma. Inactivation of the APC tumour suppressor gene, and the activation of K-ras, have a combined effect on increasing tissue polyamine contents due to increased synthesis and decreased catabolism of the polyamines. Pharmacological strategies for suppressing ODC (e.g. the enzyme-activated inhibitor alpha-difluoromethylornithine) and activating SSAT (e.g. NSAIDs) are potent inhibitors of intestinal carcinogenesis in rodent models. Clinical trials combining these classes of agent in humans with risk factors for colon cancer are in progress. PMID- 12653646 TI - Polyamine analogues as anticancer drugs. AB - Just over 30 years ago, the late Diane Russell published the first in a series of papers linking polyamines and cancer. These early studies led to a flurry of research activity in the polyamine field that continues to this day attempting to identify a role for the polyamines in cancer development, treatment and/or prevention. The recognition that polyamines are critical for the growth of cancer cells, and consequently the identification of their metabolic pathways as a target for therapeutic intervention, led to the development of a number of useful inhibitors of polyamine biosynthesis. Arguably the most significant addition to the polyamine field in the last 30 years was the synthesis of alpha difluoromethylornithine (DFMO), which is being tested currently as a cancer chemopreventative agent in man and is used also as a highly effective trypanocidal agent. Although an extremely useful tool experimentally, DFMO has been disappointing in clinical trials with little therapeutic efficacy. Despite this setback, the polyamine pathway is still considered a viable target for chemotherapeutic intervention. This has led to the development of the polyamine analogues as multifunctional inhibitors that will produce inhibition of tumour cell growth, polyamine depletion and optimum therapeutic efficacy. PMID- 12653647 TI - Polyamines and novel polyamine conjugates interact with DNA in ways that can be exploited in non-viral gene therapy. AB - As a part of our continuing studies on 'Polyamines and their role in human disease' we are investigating how polyamines, and especially how novel polyamine conjugates, interact with DNA. We are studying how these conjugates interact with circular plasmids in order to produce nanometre-sized particles suitable for transfecting cells. Our considerations of structure--activity relationships (SAR) within naturally occurring and synthetic polyamines have shown the significance of the inter-atomic distances between the basic nitrogen atoms. As these atoms are typically fully protonated under physiological conditions, they exist in equilibrium as polyammonium ions. The covalent addition of a lipid moiety, typically one or two alkyl or alkenyl chains, or a steroid, allows much greater efficiency in DNA condensation and in the cellular transfection achieved. Thus efficient DNA condensation and subsequently drug delivery (i.e. with DNA as the drug) can be brought about using novel polyamine conjugates. Taking further advantage of the functionalization of specific steroids (e.g. cholesterol and certain bile acids), we have designed and prepared novel fluorescent molecular probes as tools to throw light on the problematic steps in non-viral gene delivery which still impede efficient gene therapy. Thus, the current aims of our research are to understand, design and prepare small-molecule lipopolyamines for non-viral gene therapy (NVGT). The rational design and practical preparation of non-symmetrical polyamine carbamates and amides, based on steroid templates of cholesterol and the bile acid lithocholic acid as the lipid moiety, provides fluorescent molecular probes that condense DNA. These novel lipopolyamine conjugates mimic the positive charge distribution found in the triamine spermidine and the tetra-amine spermine alkaloids. After optimizing their SAR, these fluorescent probes will be useful in monitoring gene delivery in NVGT. PMID- 12653648 TI - The biological activities of new polyamine derivatives as potential therapeutic agents. AB - A number of polyamine derivatives have demonstrated potential as therapeutic agents. For example, 1,12-bisethylspermine and bisnaphthalimide (elinafide) are currently in phase I clinical trials for the treatment of certain cancers. Here, the biological activities of two new groups of polyamine derivative, namely the oxa-polyamines and the bisnaphthalimides, are presented. The most active compounds in the oxa-polyamine and bisnaphthalimido series possessed IC(50) values of 2.93 and 1.38 microM, respectively, against MCF7 cells after 48 h of exposure. The structure-relationship activities of each group of compounds are discussed. Bisnaphthalimido compounds are DNA-binding agents. Addition of the bisnaphthalimides PK3, PK4, PK5, PK6 and PK7, at a concentration of 10 microM, to the calf thymus DNA duplex increased the T (m) of DNA by 11.55+/-0.56, 14.545+/ 1.59, 6.23+/-2.45, 12.56+/-1.84 and 16.45+/-0.39 degrees C respectively. With the exception of PK5, all compounds bind to DNA by intercalation as judged by effect of compounds on DNA mobility. Ethidium bromide displacement assay showed that all the compounds have significant affinity for calf thymus DNA (the drug concentration required to reduce the fluorescence of initially DNA-bound ethidium bromide by 50%, C(50), was 1.21-17.33 microM). The order of DNA-binding strength was PK4 > PK3 > PK7 > PK6 > PK5. In HL-60 promyelocytic leukaemia cells, oxa polyamine and bisnaphthalimido treatment resulted in a decline in cell proliferation and viability. The assays performed suggested that apoptosis was not the principal cell death mechanism involved in oxa-polyamine cytotoxicity. In contrast, HL-60 cell death induced by the bisnaphthalimido series was characterized by early exposure of phosphatidylserine exclusive from membrane damage, elevated caspase-3 activity, increased DNA instability and, ultimately, DNA fragmentation. Thus the principal cytotoxic members of the bisnaphthalimido series appear to induce apoptosis. PMID- 12653649 TI - Turnover of trypanosomal ornithine decarboxylases. AB - Interestingly, there is a major difference in turnover rate between ornithine decarboxylases (ODCs) from various trypanosomatids. ODCs from Trypanosoma brucei and Leishmania donovani are both stable proteins, whereas ODC from Crithidia fasciculata is a metabolically unstable protein in the parasite. C. fasciculata ODC is also rapidly degraded in mammalian systems, whereas the closely related L. donovani ODC is not. The degradation of C. fasciculata ODC in the mammalian systems is shown to be dependent on a functional 26 S proteasome. However, in contrast to the degradation of mammalian ODC, the degradation of C. fasciculata ODC does not involve antizyme. Instead, it appears the degradation of C. fasciculata ODC may be associated with poly-ubiquitination of the enzyme. PMID- 12653650 TI - Polyamine biosynthetic enzymes as drug targets in parasitic protozoa. AB - Molecular, biochemical and genetic characterization of ornithine decarboxylase, S -adenosylmethionine decarboxylase and spermidine synthase establishes that these polyamine-biosynthetic enzymes are essential for growth and survival of the agents that cause African sleeping sickness, Chagas' disease, leishmaniasis and malaria. These enzymes exhibit features that differ significantly between the parasites and the human host. Therefore it is conceivable that exploitation of such differences can lead to the design of new inhibitors that will selectively kill the parasites while exerting minimal, or at least tolerable, effects on the parasite-infected patient. PMID- 12653651 TI - Polyamine metabolism in the Microsporidia. AB - Members of the phylum Microspora are all obligate intracellular parasites. Little is known concerning metabolic pathways in these parasites, some of which pose serious problems in immunocompromised patients. We investigated polyamine metabolism in the systemic pathogen Enterocytozoon cuniculi using intact pre emergent spores, and cell-free preparations. We found both polyamine synthetic and interconversion pathways to be operative, as evidenced by conversion of ornithine into polyamines, and production of spermidine from spermine by pre emergent spores. Recent developments in the antitumour field have highlighted the ability of bis-ethylated polyamine analogues to reduce polyamine levels and block growth of tumour cells. In light of enhanced polyamine uptake in Enc. cuniculi, we have begun to study bis-aryl 3-7-3 and bis-ethyl oligoamine analogues as leads for chemotherapy of microsporidia. PMID- 12653653 TI - Modern approaches to drug discovery and design: setting the scene. AB - The raison d'itre for the drug discovery and development process is to provide safe and effective treatments for diseases. Bringing a new drug to market, however, is a time-consuming and expensive process and it remains an imperative for drug companies that they identify ways in which they can accelerate the identification of potential targets and their screening and development in order to maintain a competitive edge. Successful drug discovery efforts include biochemical, biophysical, genetic and immunological approaches, targeting such processes as signal transduction, cell cycle control, apoptosis, gene regulation and metastasis. As the number of these biological targets increases, reliance on bioinformatics and chemoinformatics to improve decision making, by identifying characteristics of successful drugs and sharing knowledge gained within the scientific community, has become a burgeoning area in the post-genomic era of drug discovery. PMID- 12653652 TI - Translational regulation of the plant S-adenosylmethionine decarboxylase. AB - It is becoming apparent that control of protein synthesis by metabolites is more common than previously thought. Much of that control is exerted at the level of initiation of mRNA translation, orchestrated by upstream open reading frames (uORFs) and RNA secondary structure. S-Adenosylmethionine decarboxylase (AdoMetDC) is a key enzyme in polyamine biosynthesis and both mammalian and plant AdoMetDCs are translationally regulated by uORFs in response to polyamine levels by distinct mechanisms. PMID- 12653654 TI - Genomics in target and drug discovery. AB - Genomics-based discovery of novel therapeutic drug targets requires the design of well-controlled biological or pharmacological experiments with experimental questions and hypotheses that relate to the therapeutic area of interest. This will aid the validation level of differentially expressed genes and hence facilitate the de-selection of the genes that are identified in microarray experiments. We here provide an example of how this approach is followed in the manipulation of human macrophage foam cells towards the discovery of novel drug targets for treatment of atherosclerosis. PMID- 12653655 TI - The role of therapeutic antibodies in drug discovery. AB - The last 5 years have seen a major upturn in the fortune of therapeutic monoclonal antibodies (mAbs), with nine mAbs approved for clinical use during this period and more than 70 now in clinical trials beyond phase II. Sales are expected to reach $4 billion per annum worldwide in 2002 and $15 billion by 2010. This success can be related to the engineering of mouse mAbs into mouse/human chimaeric antibodies or humanized antibodies, which have had a major effect on immunogenicity, effector function and half-life. The issue of repeated antibody dosing at high levels with limited toxicity was essential for successful clinical applications. Emerging technologies (phage display, human antibody-engineered mice) have created a vast range of novel, antibody-based therapeutics, which specifically target clinical biomarkers of disease. Modified recombinant antibodies have been designed to be more cytotoxic (toxin delivery), to enhance effector functions (bivalent mAbs) and to be fused with enzymes for prodrug therapy and cancer treatment. Antibody fragments have also been engineered to retain specificity and have increased the penetrability of solid tumours (single chain variable fragments). Radiolabelling of antibodies has now been shown to be effective for cancer imaging and targeting. This article focuses on developments in the design and clinical use of recombinant antibodies for cancer therapy. PMID- 12653656 TI - A new paradigm for drug discovery: integrating clinical, genetic, genomic and molecular phenotype data to identify drug targets. AB - Application of statistical genetics approaches to variations in mRNA transcript abundances in segregating populations can be used to identify genes and pathways associated with common human diseases. The combination of this genetic information with gene expression and clinical trait data can also be used to identify subtypes of a disease and the genetic loci specific to each subtype. Here we highlight results from some of our recent work in this area and further explore the many possibilities that exist in employing a more comprehensive genetics and functional genomics approach to the functional annotation of genomes, and in applying such methods to the validation of targets for complex traits in the drug discovery process. PMID- 12653657 TI - A study of mitochondrial DNA mutations in peripheral lymphocytes in an aging cohort. AB - Somatic mutation in the mitochondrial genome occurs much more rapidly than in the nuclear genome and is a feature, possibly contributory, of the aging of cells and tissues. Identifying mitochondrial sequence changes in blood DNA of elderly subjects may provide a maker for the epigenetic changes of mitochondrial DNA known to occur in tissues with lower cellular turnover, and would also have implications for immunosenescence. No large-scale epidemiological studies have been reported previously. In this study we have established long-PCR banks of the mitochondrial genome from peripheral lymphocytes for an elderly cohort of 716 individuals with a range of measured aging phenotypes, and we have established assays for three widely reported mutations: the 4977 bp and 8048 bp deletions and point mutation A3243G. No individuals were identified with detectable heteroplasmy for these changes. Implications for tissue and population prevalence are discussed. The mitochondrial long-PCR DNA banks established will be useful for a wide range of studies of somatic mutation and of germline haplotypes in relation to aging. PMID- 12653658 TI - Effects of age on antibody affinity maturation. AB - The elderly are more susceptible to infectious diseases. Mortality and morbidity from infections increase sharply over the age of 65 years. At the same time, the efficacy of vaccinations in the elderly is decreased. The elderly also have an increased incidence of cancer and inflammatory diseases. All the above indicate an age-related dysregulation of the immune system. Evidence suggests that the change in the humoral immune response with age is a qualitative rather than a quantitative one, i.e. it is the affinity and specificity of the antibody that changes, rather than the quantity of antibody produced. There are a number of possible causes of this failure, one of which is a defect in the mechanism of hypermutation of immunoglobulin genes. We have studied individual clonal responses within germinal centres of spleen and Peyer's patches in young and old patient groups. Our results indicate that there is no difference in the actual mechanism of hypermutation with age. There are, however, differences that are due either to a change in selection processes or to a change in the founder cells available for activation. PMID- 12653659 TI - Effect of age on susceptibility to post-traumatic infection in the elderly. AB - Previous work has demonstrated an age-related decline in neutrophil function, including a decline in phagocytic capacity, with age in healthy individuals. This decline in function may contribute to increased susceptibility to bacterial infections in the elderly population. The present study has investigated the effects of age on susceptibility to infection and neutrophil function in elderly humans following mild trauma. Specifically, we have measured neutrophil function in 44 patients, all of whom had no significant co-morbidity, were over 65 years old (mean age 82.5 years) and had sustained a fractured neck of femur. We obtained neutrophils and examined the process of microbial engulfment by phagocytosis and the bactericidal mechanism of superoxide production. In the 5 week period after trauma, almost half of the elderly trauma patients succumbed to bacterial or fungal infection, with a predominance of chest and urinary tract infections. When examining neutrophil function, a decline in superoxide production was observed in neutrophils from the elderly trauma group at the time of hip fracture when compared with those from healthy elderly controls, and this was maintained 5 weeks after trauma. This was accompanied by an age-related reduction in phagocytic function during this period. We propose that trauma and an age-related decline in neutrophil function combine to decrease the immune response to bacteria in the elderly. PMID- 12653660 TI - Critique of the demographic evidence for 'late-life non-senescence'. AB - Although the Gompertz formula accurately describes observed mortality distributions over most of their extent, their 'tail' is much longer than that of a Gompertz curve fitted to the whole data set. A simple candidate explanation is that the longest-lived subset of any population will necessarily be enriched in individuals that age more slowly than the average of that population. However, some investigators have suggested that, instead, individuals actually cease to senesce after a certain age. Here, using a new approach to determining the best fit degree of heterogeneity in the Gompertz slope parameter, it is shown that observed distributions can in fact be fit quite accurately by purely 'heterogeneous Gompertz' curves. Either explanation may therefore be correct. PMID- 12653661 TI - Skeletal muscles of aged male mice fail to adapt following contractile activity. AB - Skeletal muscle adapts rapidly following exercise by the increased production of heat-shock proteins (HSPs). The aim of this study was to examine the ability of muscle from adult and aged mice to produce HSPs following non-damaging exercise. Adult and aged B6XSJL mice were anaesthetized and their hind limbs were subjected to isometric contractions. At different time points, muscles were analysed for HSP production by Western and Northern blotting and by electrophoretic mobility shift assay. HSP protein and mRNA levels in muscles from adult mice increased significantly following exercise. This was not evident in muscles of aged mice. In contrast, binding of the transcription factor heat-shock factor 1 (HSF1) was not grossly altered in muscles of aged mice compared with adult mice. The data suggest that the inability of muscles of aged mice to produce HSPs appears to be due to alterations during gene transcription. PMID- 12653662 TI - Centenarians as a model for healthy aging. AB - For over 10 years we have studied centenarians as a model to address the biological basis of aging and longevity, with particular attention to immunology and genetics. The most important findings can be summarized as follows. (i) Human immunosenescence represents a complex remodelling, whereby clonotypical immunity deteriorates, while ancestral, innate immunity is largely preserved. (ii) Continuous exposure to antigens causes a lifelong, chronic antigenic stress, which is responsible, together with the involution of the thymus, for the accumulation of memory/effector T cells and the exhaustion of naive T cells. (iii) Aging is characterized by a peculiar chronic inflammatory status that we propose to call 'inflammaging', which appears to be under genetic control, is detrimental for longevity and is more evident in men than in women. Inflammaging, i.e. the up-regulation of a variety of anti-stress responses at the cellular and molecular level, is the consequence of the ability of the body to adapt to and counteract the effects of a variety of stressors, which causes the accumulation of molecular and cellular scars. Inflammaging is considered the common and most important driving force of age-related pathologies, such as neurodegeneration, atherosclerosis, diabetes and sarcopenia, among others, all of which share an inflammatory pathogenesis. (iv) Possible strategies to counteract the major effects of immunosenescence and inflammaging, such as the systematic reduction of the lifelong antigenic load, the elimination of chronic infections, thymic rejuvenation and preventative treatment with anti-inflammatory drugs in people with a pro-inflammatory genotype, are envisaged. PMID- 12653663 TI - Osteoporosis: pathogenesis and clinical intervention. AB - Osteoporosis is a very common disorder and much has been learnt in recent years about the many pathogenic processes that contribute to bone loss and fragility. Drug treatments are now available to prevent bone loss and reduce fracture, and there are prospects for modifying some of the pathogenic processes themselves. In common with other structures, the tissues of the musculoskeletal system undergo many changes with aging, and some of the commonest skeletal disorders are seen in the elderly. The changes in bone lead to osteoporosis and fractures, whereas muscle changes (sarcopenia) contribute to frailty, and changes in cartilage lead to osteoarthritis. PMID- 12653664 TI - An interaction map of proteasome subunits. AB - Despite the central role of the 26 S proteasome in eukaryotic cells, many facets of its structural organization and functioning are still poorly understood. To learn more about the interactions between its different subunits, as well as its possible functional partners in cells, we performed, with Marc Vidal's laboratory (Dana-Farber Cancer Institute, Boston, MA, U.S.A.), a systematic two-hybrid analysis using Caenorhaditis elegans 26 S proteasome subunits as baits [Davy, Bello, Thierry-Mieg, Vaglio, Hitti, Doucette-Stamm, Thierry-Mieg, Reboul, Boulton, Walhout et al. (2001) EMBO Rep. 2, 821-828]. A pair-wise matrix of all subunit combinations allowed us to detect numerous possible intra-complex interactions, among which some had already been reported by others and eight were novel. Interestingly, four new interactions were detected between two ATPases of the 19 S regulatory complex and three alpha-subunits of the 20 S proteolytic core. Possibly, these interactions participate in the association of these two complexes to form the 26 S proteasome. Proteasome subunit sequences were also used to screen a cDNA library to identify new interactors of the complex. Among the interactors found, most (58) have no clear connection to the proteasome, and could be either substrates or potential cofactors of this complex. Few interactors (7) could be directly or indirectly linked to proteolysis. The others (12) interacted with more than one proteasome subunit, forming 'interaction clusters' of potential biological interest. PMID- 12653665 TI - Proteasomal interactors control activities as diverse as the cell cycle and glutaminergic neurotransmission. AB - The six regulatory non-redundant ATPases in the base of the 19 S regulator of the 26 S proteasome belong to the AAA superfamily of ATPases. Yeast two-hybrid genetic screens, biochemical analyses and cell biological studies have identified and characterized new interactors of the human S6 (rpt3) and S8 (rpt6) ATPases of the 19 S regulator of the 26 S proteasome. The S6 ATPase interacts with gankyrin. This protein is found in purified human 26 S proteasomes and in a smaller complex(es) containing CDK4 and free S6 ATPase. Gankyrin overexpression causes the phosphorylation of the retinoblastoma protein (pRb) and the release of E2F transcription factor to trigger the expression of DNA synthesis genes. Gankyrin is oncogenic in nude mice and is overexpressed in hepatocellular carcinoma cells (HCCs). The S8 ATPase interacts with members of the large Homer-3 protein family. There are three Homer genes; the Homer 1 and 2 gene products control trafficking and calcium-store-related functions of metabotropic glutamate receptors (e.g. mGluR1alpha). Homer-3A11 by binding to the S8 ATPase brings mGluR1alpha to the 26 S proteasome for degradation. The degradation of mGluR1alpha is blocked by proteasomal inhibitors and by overexpression of the N-terminus of Homer which binds to the receptor. The S8 ATPase and mGluR1alpha are co-localized in Purkinje dendrites in rat cerebellum. The data are discussed in terms of the regulation of the cell cycle and glutaminergic receptor functions by the 26 S proteasome. PMID- 12653666 TI - The ubiquitin proteolytic system and pathogenesis of human diseases: a novel platform for mechanism-based drug targeting. AB - Until the early 1980s, protein degradation was a neglected research area, and scientists were mostly busy deciphering the genetic code and its translation to the proteome. Destruction of cellular proteins was thought to be a scavenger, non specific and dead-end process. Although it was known that proteins do turn over, the large extent and high specificity of the process, whereby distinct proteins have half-lives that range from a few minutes to several days, was not appreciated. The discovery of the lysosome by Christian de Duve did not change this view significantly, as it was clear that this organelle is involved mostly in the degradation of extracellular proteins, and their proteases cannot be substrate-specific. The discovery of the complex cascade of the ubiquitin pathway revolutionized the field. It is clear now that degradation of cellular proteins via the ubiquitin system is a highly complex, temporally controlled and tightly regulated process that plays major roles in a variety of basic pathways and processes during cell life and death, and in health and disease. The system is involved in targeting many cellular proteins, among them cell cycle regulators, growth- and differentiation-controlling factors, transcriptional activators, cell surface receptors and ion channels, endoplasmic reticulum proteins, antigenic proteins destined for presentation on class I MHC molecules, and abnormal/misfolded proteins. Consequently, it is involved in regulating many basic cellular processes, such as cell cycle and division, growth and differentiation, signal transduction and transcription, modulation of the secretory and endocytic pathways, the immune and inflammatory responses, and quality control. With the multitude of substrates targeted and the numerous processes involved, it is not surprising that aberrations in the pathway have been implicated in the pathogenesis of many diseases, with certain malignancies and neurodegenerative disorders being ranked among them. PMID- 12653667 TI - Protecting p53 from degradation. AB - Inactivation of the p53 function is a common event in cancer. Approx. 50% of human tumours express mutant p53 and there is evidence that in others, including many childhood tumours, p53 function is impaired in other ways. These defects on p53 function may be due to the alteration of cellular factors that modulate p53 or to the expression of viral oncoproteins. Radiotherapy and many of the chemotherapeutic drugs currently used in cancer treatment are potent activators of p53. However, most of these therapies have a serious drawback; that is, the long-term consequences of their DNA-damaging effects. Understanding the mechanisms regulating p53 stability is crucial for the development of new strategies to activate p53 non-genotoxically. Here we describe the effect of a potent activator of the p53 response, the nuclear export inhibitor leptomycin B, on Mdm2 degradation and we provide evidence for the oligomerization of the p14ARF tumour suppressor and Mdm2 inhibitor in response to oxidative stress. PMID- 12653668 TI - Target-based agents against ErbB receptors and their ligands: a novel approach to cancer treatment. AB - The ErbB receptors and their cognate ligands that belong to the epidermal growth factor (EGF) family of peptides are involved in the pathogenesis of different types of carcinomas. In fact, the ErbB receptors and the EGF-like growth factors are frequently expressed in human tumors. These proteins form a complex system that regulates the proliferation and the survival of cancer cells. Therefore, ErbB receptors and their ligands might represent suitable targets for novel therapeutic approaches in human carcinomas. In this regard, different target based agents that are directed against the ErbB receptors have been developed in the past two decades. One of these compounds, the humanized anti-ErbB-2 monoclonal antibody trastuzumab has been approved for the treatment of patients with metastatic breast cancer. The anti-EGF receptor (EGFR) antibody C225, as well as EGFR tyrosine kinase inhibitors ZD1839 and OSI-774 are currently in phase III clinical development. Several other ErbB tyrosine kinase inhibitors are in phase I/II studies. These compounds have generally been shown to have an acceptable toxicity profile and promising anti-tumor activity in heavily pretreated patients. The mechanisms of action of these compounds, as well as the potential therapeutic strategies to improve their efficacy are discussed in this review with particular regard to the combinations of anti-ErbB agents with cytotoxic drugs, or combinations of different ErbB-targeting agents. PMID- 12653669 TI - Endometrial cancer: experimental models useful for studies on molecular aspects of endometrial cancer and carcinogenesis. AB - There is definitely a need for the development of new drugs for the treatment and cure of endometrial cancer. In addition there are various new drugs or phyto remedies under development which are intended for use in the treatment and prevention of breast cancer, for the treatment of menopausal symptoms and for hormone replacement therapy. The efficacy of novel drugs targeting steroid receptors in endometrial cancers has to be evaluated and the safety of other endocrine measures on endometrial cancers or on endometrial carcinogenesis has to be assessed. For these experimental purposes five main classes of experimental models are available: spontaneous endometrial tumorigenesis models in inbred animals (Donryu rats, DA/Han rats, BDII/Han rats), inoculation tumors from chunks of tumors (rat EnDA-tumor, human EnCa 101 tumor) or from inoculated tumor cell lines (rat RUCA-I cells, human Ishikawa and ECC-1 cells), developmental estrogenic exposure or chemical carcinogen exposure of CD-1 and ICR mice, transgenic approaches such as mice heterozygous regarding the tumor suppressor gene PTEN (pten(+/-)-mice) and endometrial tumor cell lines cultured under conditions promoting in vivo-like morphology and functions e.g. cell culture on reconstituted basement membrane. Although the number of models is comparatively small, most aspects related to functions of estrogenic or gestagenic substances are assessable, particularly if various experimental models are combined. Whereas models based on human endometrial adenocarcinoma cells are widely used, the properties and advantages of animal-derived models have mainly been ignored so far. PMID- 12653671 TI - New paradigms in adjuvant systemic therapy of breast cancer. AB - Since the initial studies of adjuvant therapy in the 1970s, it has become increasingly clear that chemotherapy and hormone therapy have had a substantial effect on the survival of women with early breast cancer. It was originally assumed that only women with high-risk features would derive benefit from adjuvant therapy, but it is now apparent from numerous studies that adjuvant therapy improves survival in all subgroups of women with invasive breast cancer, although the absolute benefit varies depending on tumor stage and other prognostic features. Considerable progress has been made in elucidating effective adjuvant therapy regimens, but there continue to be many unanswered questions that are being addressed in ongoing clinical trials of adjuvant hormone therapy and chemotherapy. This paper reviews the current paradigms in adjuvant therapy, the published data that have affected current practice patterns, and the current controversies. PMID- 12653670 TI - ATP binding cassette transporters and drug resistance in breast cancer. AB - Resistance to chemotherapy is a critical issue in the management of breast cancer patients. The nature of clinical drug resistance is likely to be multifactorial. However, in the last decade considerable attention has been dedicated to the role played by membrane transporter proteins belonging to the ATP binding cassette protein superfamily, and in particular by the MDR1 product P-glycoprotein (Pgp) and the multidrug resistance protein (MRP1). Heterogeneity of results is a common feature of studies evaluating the expression and prognostic role of these proteins, due to both methodological and biological factors. Nonetheless, Pgp and MRP1 are detected in a significant proportion of untreated breast cancers (on average 40 and 50% respectively, by immunohistochemistry), without a clear and consistent association with cancer stage. Exposure to chemotherapy increases the expression of both proteins. In vitro studies on primary cultures of breast cancer cells obtained at surgery consistently show an association between Pgp (protein) or MDR1 (mRNA) expression and resistance to chemotherapy. However, the correlation with clinical drug resistance is not as well defined. A stronger association of Pgp/MDR1 with response rates has been observed when expression or an increase in expression are detected immediately following chemotherapy. Correlations with prognosis appear more evident in studies using immunohistochemistry, in adjuvant and neoadjuvant settings. Evidence of clinical reversal of drug resistance by verapamil suggests a functional role of Pgp in drug resistance, although the significance of the evidence is generally weakened by poor trial designs. Future studies should take into account the multifactorial nature of drug resistance in breast cancer and use standardized approaches with adequate controls. Expression studies should be complemented by well-designed trials of drug-resistance reversal using target-specific chemosensitizing agents, and relating the results to the levels of expression of the target proteins. PMID- 12653672 TI - Sequential steroid hormone receptor measurements in primary breast cancer with and without intervening primary chemotherapy. AB - The objective of this analysis was to determine the accuracy of steroid receptor measurement in large core needle biopsies compared with surgically removed specimens and the influence of preoperative chemotherapy on hormone receptor status. We consecutively performed 722 large core needle biopsies in palpable lesions of the breast. The diagnosis of breast cancer was confirmed upon biopsy in 450 patients; 236 women underwent immediate surgery, and 214 patients received preoperative chemotherapy. We assessed estrogen (ER) and progesterone receptor (PR) in biopsy tissue and surgically removed specimens and calculated accuracy, sensitivity, specificity, the weighted kappa value and Spearman's rank correlation. The modulation of steroid receptor status in preoperatively treated patients was tested by Cochran-Mantel-Haenszel statistics. The accuracy of ER evaluation in the biopsy material of patients without intervening chemotherapy was 91%, sensitivity and specificity were 94% and 80% respectively. Accuracy, sensitivity and specificity were 86% in patients treated preoperatively. In terms of PR assessment, we obtained slightly inferior results: accuracy, sensitivity and specificity were 80%, 73% and 85% respectively in patients without preoperative treatment, and 79%, 48% and 92% respectively in patients undergoing preoperative therapy. Following preoperative chemotherapy, patients showed a significant increase in ER-negative (P=0.02) and PR-negative (P=0.0005) measurements. We have concluded from our results that ER and PR receptor measurement in core needle biopsy is a reliable basis in clinical practice for selecting patients for neoadjuvant endocrine treatment. Preoperative cytotoxic chemotherapy induced a significant extent of variation in the steroid receptor expression of breast cancer cells. PMID- 12653673 TI - Adrenal ganglioneuroma in a patient presenting with severe hypertension and diarrhea. AB - Ganglioneuromas (GNs) are neural crest cell-derived tumors and rarely occur in the adrenal gland. There are presently no markers that can reliably distinguish benign and malignant neuroendocrine tumors. Here we describe a 63-year-old woman who developed sudden chest pain and hypertension combined with increased stool frequency. An incidental adrenal mass 5 cm in size with a bright signal on T2 weighted magnetic resonance imaging was discovered. Biochemical evaluation and (131)I-metaiodobenzylguanidine (MIBG) scintigraphy were negative. Histopathological examination revealed a mature adrenal GN. Neuroblastoma, the immature form of a GN, is known for deletions on chromosomal locus 1p36, and adrenal tumors frequently show allele loss on 17p. To further elucidate the histo and pathogenesis of adrenal GN, we performed loss of heterozygosity studies on chromosomal loci 1p34-36 and 17p13 (the p53 gene locus) after careful microdissection of tumor and normal tissue. We did not detect allelic losses at these loci with the informative polymorphic markers used, suggesting that these loci are not involved in tumorigenesis. In addition, immunohistochemical investigation of the GN was positive for vasoactive intestinal peptide, a hormone commonly expressed in ganglion cells. We suggest that in our patient with an adrenal GN, the combination of biochemical, scintigraphic, molecular, immunohistochemical, and histopathological findings are all consistent with the benign morphology of this tumor. PMID- 12653674 TI - Contrasting neurovascular findings in chronic orthostatic intolerance and neurocardiogenic syncope. AB - Simple faint (neurocardiogenic syncope) and postural tachycardia syndrome (POTS) characterize acute and chronic orthostatic intolerance respectively. We explored the hypothesis that vascular function is similar in the two conditions. We studied 29 patients with POTS and compared them with 20 patients with neurocardiogenic syncope who were otherwise well, and with 15 healthy control subjects. We measured continuous heart rate, respiration and blood pressure, and used venous occlusion strain gauge plethysmography to measure calf and forearm blood flow, peripheral arterial resistance, peripheral venous resistance and venous pressure ( P (v)). Upright tilt was performed to 70 degrees for 10 min, during which calf blood flow and volume were measured. Calf P (v) was increased (to 27.2+/-2.0 mmHg) in a subgroup of POTS patients, who also had increased arterial resistance (57+/-6 mmHg.ml(-1).min(-1).100 ml(-1) tissue), increased venous resistance (2.4+/-0.3 mmHg.ml(-1).min(-1).100 ml(-1) tissue), and decreased peripheral flow (1.0+/-0.2 ml.min(-1).100 ml(-1) tissue) in the calf; other POTS patients with a normal P (v) had decreased arterial resistance (18+/-2 mmHg.ml(-1).min(-1).100 ml(-1) tissue) and increased blood flow (3.8+/-0.3 ml.min(-1).100 ml(-1) tissue). Syncope patients were not different from controls ( P (v)=11.4+/-0.5 mmHg; calf flow=3.1+/-0.2 ml.min(-1).100 ml(-1) tissue; arterial resistance=27+/-2 mmHg.ml(-1).min(-1).100 ml(-1) tissue; venous resistance=1.2+/-0.3 mmHg.ml(-1).min(-1).100 ml(-1) tissue). When upright, syncope patients and control subjects had similar increases in heart rate and calf volume, stable blood pressure, and decreases in blood flow. POTS patients had markedly increased heart rate and calf blood flow, unstable blood pressure, and pooling in the lower extremities, regardless of subgroup. We conclude that peripheral vascular physiology in patients with POTS is abnormal, in contrast with normal peripheral vascular physiology in neurocardiogenic syncope. PMID- 12653676 TI - High-dose heparin fails to improve acute lung injury following smoke inhalation in sheep. AB - Thrombin is involved in various inflammatory responses. In sepsis, coagulation abnormalities are major complications. Acute lung injury is one of the most life threatening problems that can result from sepsis. We hypothesized that high-dose heparin might be effective in attenuating acute lung injury in our sepsis model. Female sheep ( n =16) were surgically prepared for the study. After a tracheotomy, 48 breaths of cotton smoke (<40 degrees C) were insufflated into the airway. Afterwards, live Pseudomonas aeruginosa (5 x 10(11) colony-forming units) bacteria were instilled into the lung. All sheep were ventilated mechanically with 100% O(2), and were divided into three groups: a heparin infusion group ( n =6), a Ringer's lactate infusion group ( n =6), and a sham-injury group ( n =4; surgically prepared in the same fashion but receiving no inhalation injury or bacteria). The treatment was started 1 h after the insult, and was continued thereafter for 24 h. The dose of heparin was adjusted by monitoring to target an activated clotting time of between 300 and 400 s (baseline=approx. 150 s). Sheep exposed to lung injury presented with typical hyperdynamic cardiovascular changes, including an increased cardiac output and a fall in systemic vascular resistance. There was a decrease in the arterial partial pressure of O(2). In conclusion, high-dose heparin did not prevent lung dysfunction in this model, in which acute lung injury was induced by combined smoke and septic challenge. PMID- 12653675 TI - Cardiovascular hypertrophy in diabetic spontaneously hypertensive rats: optimizing blockade of the renin-angiotensin system. AB - The aim of the present study was to compare the antihypertrophic effects of blockade of the renin-angiotensin system (RAS), vasopeptidase inhibition and calcium channel antagonism on cardiac and vascular hypertrophy in diabetic spontaneously hypertensive rats (SHR). SHR with streptozotocin-induced diabetes were treated with one of the following therapies for 32 weeks: the angiotensin converting enzyme (ACE) inhibitor captopril (100 mg/kg); the angiotensin AT(1) receptor antagonist valsartan (30 mg/kg); a combination of captopril with valsartan; the vasopeptidase inhibitor mixanpril (100 mg/kg); or the calcium channel antagonist amlodipine (6 mg/kg). Systolic blood pressure and cardiac and mesenteric artery hypertrophy were assessed. Mean systolic blood pressure in diabetic SHR (200+/-5 mmHg) was reduced by captopril (162+/-5 mmHg), valsartan (173+/-5 mmHg), mixanpril (176+/-2 mmHg) and amlodipine (159+/-4 mmHg), and was further reduced by the combination of captopril with valsartan (131+/-5 mmHg). Captopril, valsartan and mixanpril reduced heart and left ventricle weights by approx. 10%. The combination of captopril and valsartan further reduced heart weight (-24%) and left ventricular weight (-29%). Amlodipine did not affect cardiac hypertrophy. Only mixanpril and the combination of captopril and valsartan significantly reduced mesenteric weight. The mesenteric wall/lumen ratio was reduced by all drugs, and to a greater extent by the combination of captopril and valsartan. We conclude that optimizing the blockade of vasoconstrictive pathways such as the RAS, particularly with the combination of ACE inhibition and AT(1) receptor antagonism, is associated with antitrophic effects in the context of diabetes and hypertension. In contrast, calcium channel blockade, despite similar effects on blood pressure, confers less antitrophic effects in the diabetic heart and blood vessels. PMID- 12653678 TI - Evaluation of methods to measure endothelium-dependent vasodilation. PMID- 12653680 TI - Haematological phenotypes in relation to the C1797T beta-adducin polymorphism in a Caucasian population. AB - beta-Adducin plays a role in maintaining the structural integrity of the red blood cell (erythrocyte) membrane. Moreover, beta-adducin-deficient knock-out mice show a phenotype characterized by mild anaemia and compensated haemolysis. We therefore investigated whether, in humans, common haematological phenotypes of red blood cells were associated with a polymorphism in exon 15 of the human beta adducin gene (C1797T). We studied 802 unrelated individuals and 294 families (459 parents and 609 offspring) randomly selected from a Caucasian population. We employed generalized estimating equations to allow for the non-independence of the observations within families, while controlling for co-variables. In 917 men, with adjustments applied for age, body mass index, serum total cholesterol, smoking and alcohol intake, CC homozygotes had significantly ( P =0.02) lower values for red blood cell count (4.93 x 10(12)/l compared with 4.86 x 10(12)/l), haemoglobin level (9.30 compared with 9.18 mmol/l) and haematocrit (45.0% compared with 44.4%) than T allele carriers. In the 329 men who consumed alcohol, the differences between CC homozygotes and T allele carriers were 0.13 x 10(12)/l ( P =0.02) for red blood cell count, 0.23 mmol/l ( P =0.005) for haemoglobin and 1.08% ( P =0.02) for haematocrit. In 953 women, none of these associations was significant ( P >/=0.06), regardless of alcohol intake [13.3% of women ( n =127) consmued alcohol]. In conclusion, in men consuming alcohol, the beta-adducin CC genotype was associated with lower red blood cell count, haemoglobin level and haematocrit. We hypothesize that, in CC homozygotes, alcohol consumption may unveil the greater fragility of the red blood cell membrane. This genotype may slightly potentiate the structural and functional haematological disturbances associated with alcohol intake. PMID- 12653677 TI - Comparison between the effects of mixed dyslipidaemia and hypercholesterolaemia on endothelial function, atherosclerotic lesions and fibrinolysis in rabbits. AB - We compared the impact of hypercholesterolaemia and mixed dyslipidaemia on vascular function, vascular structure and fibrinolytic balance in rabbits. To this end, vascular reactivity was studied in aortic rings from rabbits fed a control diet, a diet containing 0.5% cholesterol+14% coconut oil (mixed dyslipidaemia) or a diet containing 1% cholesterol (hypercholesterolaemia) for 12 14 weeks. Morphometric analysis of aorta was also performed and plasminogen activator inhibitor-1 (PAI-1) as well as tissue-type plasminogen activator (t-PA) plasma activities were measured. Both diets induced a similar increase in cholesterol plasma levels, although triacylglycerols (triglycerides) were increased in animals with mixed dyslipidaemia. Hypercholesterolaemia was associated with intimal thickening, reduction in acetylcholine-induced relaxation ( P <0.05) and increased vasoconstriction induced by acetylcholine+ N (G)-nitro-L arginine methyl ester (L-NAME) when compared with controls ( P <0.05). These effects were more marked ( P <0.05) in animals with mixed dyslipidaemia. Incubation with ifetroban, a thromboxane A(2)/prostaglandin H(2) receptor antagonist, increased acetylcholine-induced relaxation ( P <0.05) and reduced acetylcholine+L-NAME contraction ( P <0.05) in both diet groups. In contrast, the presence of PD 145, an endothelin (ET)(A)/ET(B) receptor antagonist, exerted these effects only in rabbits with mixed dyslipidaemia. Both hypercholesterolaemia and mixed dyslipidaemia induced a similar increase in PAI-1 and a similar decrease in t-PA plasma activities. These data suggest that hypertriglyceridaemia can increase the deleterious effects of hypercholesterolaemia on endothelial function and vascular structure. This additional harmful effect exerted by triacylglycerols on endothelial function could, in part, be mediated by ET. PMID- 12653681 TI - Clinical and electrophysiological characterization of a novel mutation (F193L) in the KCNQ1 gene associated with long QT syndrome. AB - KCNQ1 is a gene encoding an alpha subunit of voltage-gated cardiac K(+) channels, with properties similar to the slowly activating delayed rectifier K(+) current, and one of the genes causing long QT syndrome (LQTS). However, genotype-phenotype correlations of the KCNQ1 gene mutations are not fully understood. The aims of this study were to identify a mutation in the KCNQ1 gene in patients with LQTS, and to characterize the clinical manifestations and electrophysiological properties of the mutation. We screened and identified mutations by PCR, single strand conformational polymorphism analysis and DNA sequencing. We identified a novel mutation [Phe193Leu (F193L)] in the KCNQ1 gene in one family with LQTS. The patients with this mutation showed a mildly affected phenotype. The proband was a 17-year-old girl who had a prolonged QT interval. Her elder brother, father and paternal grandmother also had the mutation. None of them had any history of syncope. Sudden death was not found in this family. Next, we studied the electrophysiological characteristics of the F193L mutation in the KCNQ1 gene using the expression system in Xenopus oocytes and the two-microelectrode voltage clamp technique. Co-expression of F193L KCNQ1 with the K(+) channel minK suppressed peak (by 23.3%) and tail (by 38.2%) currents compared with those obtained by the combination of wild-type (WT) KCNQ1 and minK. Time constants of current activation in F193L KCNQ1 and F193L KCNQ1+minK were significantly slower than those of WT KCNQ1 and WT KCNQ1+minK. This electrophysiological study indicates that F193L causes less severe KCNQ1 current suppression, and thereby this mutation may result in a mildly affected phenotype. PMID- 12653682 TI - Relationships between cholesterol homoeostasis and triacylglycerol-rich lipoprotein remnant metabolism in the metabolic syndrome. AB - The dysmetabolic syndrome of insulin resistance and visceral obesity is characterized by elevated plasma concentration of triacylglycerol-rich lipoprotein (TRL) remnants that may be related to increased cardiovascular risk. Perturbed hepato-intestinal cholesterol metabolism may play a contributory role in this abnormality. We therefore investigated the association between plasma markers of cholesterol absorption and synthesis with TRL remnant metabolism in 35 men with the metabolic syndrome (MS). Plasma campesterol:cholesterol and lathosterol:cholesterol ratios were measured as estimates of cholesterol absorption and synthesis respectively. Remnant metabolism was assessed by measuring remnant-like particle-cholesterol (RLP-C), apolipoprotein (apo)B-48 and the fractional catabolic rate (FCR) of a labelled remnant-like emulsion. Compared with controls, subjects with the MS had significantly lower plasma campesterol:cholesterol ratio, but higher lathosterol:cholesterol ratio ( P <0.05). Plasma RLP-C and apoB-48 concentrations were also higher ( P <0.01) and the remnant-like emulsion FCR was lower ( P <0.05). The plasma campesterol:cholesterol ratio was inversely correlated ( P <0.05) with plasma triacylglycerols ( r =-0.346), RLP-C ( r =-0.443), apoB-48 ( r =-0.427) and plasma lathosterol:cholesterol ratio ( r =-0.366); the campesterol:cholesterol ratio was also positively correlated with the remnant-like emulsion FCR ( r =0.398, P <0.05). In multiple regression analysis, the significant correlations between plasma campesterol:cholesterol ratio and plasma triacylglycerols, RLP-C, apoB-48 and FCR of the remnant-like emulsion were independent of age, dietary energy and plasma lathosterol. Our findings suggest that in subjects with the MS alterations in cholesterol absorption and synthesis may be closely linked with the kinetic defects in TRL metabolism. PMID- 12653683 TI - Epithelial sodium channel (ENaC) subunit mRNA and protein expression in rats with puromycin aminonucleoside-induced nephrotic syndrome. AB - In experimental nephrotic syndrome, urinary sodium excretion is decreased during the early phase of the disease. The molecular mechanism(s) leading to salt retention has not been completely elucidated. The rate-limiting constituent of collecting duct sodium transport is the epithelial sodium channel (ENaC). We examined the abundance of ENaC subunit mRNAs and proteins in puromycin aminonucleoside (PAN)-induced nephrotic syndrome. The time courses of urinary sodium excretion, plasma aldosterone concentration and proteinuria were studied in male Sprague-Dawley rats treated with a single dose of either PAN or vehicle. The relative amounts of alphaENaC, betaENaC and gammaENaC mRNAs were determined in kidneys from these rats by real-time quantitative TaqMan PCR, and the amounts of proteins by Western blot. The kinetics of urinary sodium excretion and the appearance of proteinuria were comparable with those reported previously. Sodium retention occurred on days 2, 3 and 6 after PAN injection. A significant up regulation of alphaENaC and betaENaC mRNA abundance on days 1 and 2 preceded sodium retention on days 2 and 3. Conversely, down-regulation of alphaENaC, betaENaC and gammaENaC mRNA expression on day 3 occurred in the presence of high aldosterone concentrations, and was followed by a return of sodium excretion to control values. The amounts of alphaENaC, betaENaC and gammaENaC proteins were not increased during PAN-induced sodium retention. In conclusion, ENaC mRNA expression, especially alphaENaC, is increased in the very early phase of the experimental model of PAN-induced nephrotic syndrome in rats, but appears to escape from the regulation by aldosterone after day 3. PMID- 12653684 TI - Vascular endothelial growth factor and tissue factor in patients with established peripheral artery disease: a link between angiogenesis and thrombogenesis? AB - Increasing evidence points towards a prothrombotic state in atherosclerosis and its manifestations, such as peripheral artery disease (PAD), which is associated with thrombosis-related complications, such as acute limb ischaemia, graft thrombosis and stroke. We hypothesized that the increased risk of thrombogenesis in PAD may be related to abnormal angiogenesis and, thus, an increased risk of future vascular disease. To test this hypothesis, we measured plasma levels of tissue factor (TF) and related levels to indices of angiogenesis, that is vascular endothelial growth factor (VEGF) and its soluble receptor sFlt-1. We studied 234 patients (145 males; mean age 68.6+/-10 years) with proven PAD (ankle brachial pressure index <0.8) and compared them with 50 healthy controls. Levels of VEGF ( P =0.001) and TF ( P =0.043) were increased in patients compared with controls. There were significant correlations between VEGF and TF levels in both patients (Spearman r =0.351, P <0.001) and healthy controls (Spearman r =0.335, P =0.017). Amongst PAD patients, levels of VEGF were related to gender, with women having higher levels than men. There was no difference in the levels of sFlt-1 between the patients and controls, or between the subgroups of patients. There were however significant correlations between the levels of sFlt-1 and TF (Spearman r =0.268, P <0.001) and between sFlt-1 and VEGF (Spearman r =0.499, P <0.001). In conclusion, patients suffering from proven PAD have higher plasma levels of TF and VEGF compared with controls, with a significant correlation between the two. This suggests a link between the hypercoagulable state in PAD and the process of angiogenesis. PMID- 12653685 TI - Evidence of impaired microvascular function in pre-eclampsia: a non-invasive study. AB - The clinical presentation of pre-eclampsia suggests that microvascular dysfunction may play a role in the maternal manifestations of the disease. Isovolumetric venous pressure ( P V(i)) is an index of microvascular function, reflecting local plasma colloid osmotic (oncotic) pressure, and is abnormal in clinical conditions with microvascular dysfunction. We hypothesized that, in pre eclampsia, post-capillary margination of neutrophils would increase post capillary resistance, and therefore P V(i). A small cumulative step strain-gauge plethysmography protocol was used to compare P V(i) in 18 women with pre eclampsia, 16 normal pregnant women and 17 non-pregnant controls. Circulating levels of vascular cell-adhesion molecule-1 (VCAM-1), intercellular cell-adhesion molecule-1 (ICAM-1) and E-selectin, and neutrophil elastase, were measured to assess endothelial and neutrophil activation respectively. P V(i) was significantly greater in the pre-eclampsia group, relative to the normal pregnant and non-pregnant controls ( P <0.001, ANOVA, for both comparisons). P V(i) was significantly lower during normal pregnancy compared with the non-pregnant controls ( P =0.001). Plasma levels of neutrophil elastase, VCAM-1, ICAM-1 and E selectin ( P =0.001) were significantly greater in the pre-eclamptics than the controls. Significant positive correlations were observed between P V(i) and neutrophil elastase ( r =0.71, P =0.001), VCAM-1 ( r =0.52, P =0.03), ICAM-1 ( r =0.67, P =0.002), E-selectin ( r =0.69, P =0.001), uric acid levels ( r =0.54, P =0.02) and haematocrit ( r =0.64, P =0.004) in pre-eclampsia. The relationship with the platelet count was negative ( r =-0.65, P =0.003). No significant correlations were observed between P V(i) and maternal age, gestational age, total protein, albumin, diastolic blood pressures, age, body mass index and infant birth mass in the normal pregnant and non-pregnant controls. These data suggest that microvascular dysfunction occurs in pre-eclampsia, and that it is related to alterations in endothelial cell and neutrophil activation. PMID- 12653686 TI - Pre-eclampsia and the microcirculation: a novel explanation. PMID- 12653687 TI - Indeterminate Chagas' disease: Trypanosoma cruzi strain and re-infection are factors involved in the progression of cardiopathy. AB - Chagas' disease is caused by Trypanosoma cruzi, which is transmitted by reduviid bugs. The World Health Organization has estimated that about 16-18 million people in the Americas are infected, and that more than 100 million are at risk. In the present study we have used a murine model to analyse if particular T. cruzi strains (Tulahuen strain and SGO-Z12 isolate from a chronic patient) and/or re infection may determine, during the indeterminate phase of experimental Chagas' disease, changes that could explain the different evolution of cardiac lesions. Re-infected mice reached higher parasitaemias than those infected for the first time. The survival in the indeterminate phase of mice infected with Tulahuen strain was 50.0%, while the SGO-Z12-infected group presented a significantly higher survival rate (77.1%; P <0.01). The SGO-Z12-re-infected group showed a survival rate (70.9%) significantly higher than that of the Tulahuen-re-infected group (37.0%; P <0.01). Electrocardiographic abnormalities were found in 66% of Tulahuen-infected mice, while in SGO-Z12-infected group such abnormalities were found in only 36% of animals ( P <0.01). The two groups exhibited similar percentages of electrocardiographic dysfunction on re-infection, although intraventricular blocks were more frequent in Tulahuen-re-infected mice ( P <0.01). Hearts from infected or re-infected mice with either parasite showed mononuclear infiltrates. The SGO-Z12-re-infected and Tulahuen-re-infected groups exhibited a significantly diminished affinity ( P <0.05) and a significantly increased density ( P <0.05) of cardiac beta-adrenergic receptors compared with the infected and non-infected groups. The indeterminate phase of Chagas' disease is defined as a prolonged period that is clinically silent, but the present findings show that different T. cruzi strains and re-infection are able to alter the host-parasite equilibrium, and these factors may be responsible for inducing progressive cardiopathy. PMID- 12653688 TI - Cyclical mechanical stretch enhances angiopoietin-2 and Tie2 receptor expression in cultured human umbilical vein endothelial cells. AB - Endothelial cells are essential for neovascularization. Angiopoietins and Tie receptors are required for a normal vasculature. How cyclical mechanical stretch affects the expression of components of the angiopoietin system is not known. In this study, we investigated the regulation of angiopoietins and Tie receptors by cyclical mechanical stretch in cultured human umbilical vein endothelial cells (HUVECs). HUVECs grown on a flexible membrane base were stretched by vacuum to 20% elongation, at 60 cycles/min. The levels of angiopoietin-2 protein began to increase as early as 2 h after stretch was initially applied, reached a maximum of 2.7-fold over the control value by 6 h. The Tie2 receptor protein showed the same pattern as Ang-2. These increases in angiopoietin-2 and Tie2 receptor proteins at 6 h were blocked by the addition (30 min before stretch) of the protein kinase C inhibitor Go6976 (16 nM) or the tyrosine kinase inhibitor herbimycin A (24 microM). Similar to protein expression, the levels of angiopoietin-2 and Tie2 receptor mRNAs in HUVECs increased 3.1-fold and 2.5-fold respectively after stretch for 6 h. These increases were also blocked by Go6976 or herbimycin A. Cyclical mechanical stretch increased (and Go6976 or herbimycin A abrogated these increases) the immunohistochemical labelling of angiopoietin-2 and Tie2 receptor after a 6 h stretch. The levels of angiopoietin-1 and Tie1 receptor proteins, mRNAs and immunohistochemical staining were unaffected by cyclical mechanical stretch. Thus cyclical mechanical stretch activates the expression of angiopoietin-2 and the Tie2 receptor, but not angiopoietin-1 or the Tie1 receptor, in cultured HUVECs. This mechanical effect is probably mediated by the tyrosine kinase and protein kinase C pathways. PMID- 12653689 TI - Natriuretic peptides maintain sodium homoeostasis during chronic volume loading post-myocardial infarction in sheep. AB - The impaired ability to excrete sodium is a key feature of established congestive heart failure and is also apparent in asymptomatic left ventricular (LV) impairment. However, few studies have examined responses to chronic volume loading immediately post-myocardial infarction (MI). Experimental MI was induced in six sheep by thrombogenic coil coronary artery occlusion, and resulted in significant LV dysfunction with reduced LV ejection fraction ( P =0.001) and subsequent remodelling (increased LV volumes, P =0.015). Chronic volume loading with 2, 3 and 4 litres/day intravenous saline (each for 7 days) showed no evidence of renal sodium or volume retention in sheep with experimental MI compared with six normal control sheep. Plasma levels of brain natriuretic peptide (BNP), N-terminal pro-BNP and cGMP (all P <0.05) were higher in the MI group compared with normal control sheep. There were no differences in haemodynamics, body mass or renin-aldosterone levels between groups. This study provides evidence that natriuretic peptides play a pivotal role in preserving volume/electrolyte balance in the early stages of post-MI cardiac dysfunction. PMID- 12653690 TI - DC-SIGN (dendritic cell-specific ICAM-grabbing non-integrin) and DC-SIGN-related (DC-SIGNR): friend or foe? AB - C-type lectins are calcium-dependent carbohydrate-binding proteins with a wide range of biological functions, many of which are related to immunity. DC-SIGN (dendritic cell-specific ICAM-grabbing non-integrin, where ICAM is intercellular adhesion molecule) is a recently described mannose-specific C-type lectin expressed by dendritic cells. Dendritic cells are potent antigen-presenting cells capable of activating T-lymphocytes. DC-SIGN, which is expressed by dendritic cells, binds to ICAM-3 on T-lymphocytes, therefore playing an important role in the activation of T-lymphocytes. DC-SIGN can also bind HIV, and the virus may remain bound to DC-SIGN for protracted periods. DC-SIGN may deliver bound HIV to permissive cell types, mediating infection with high efficiency. A closely related C-type lectin, DC-SIGN-related molecule (DC-SIGNR) has also been described. DC-SIGNR is expressed by restricted subsets of endothelial cells, but has similar ICAM-3 and HIV-binding properties to DC-SIGN. This review describes the mapping of DC-SIGN and DC-SIGNR to chromosome 19p13.3 adjacent to the previously described C-type lectin, CD23 [the low-affinity receptor for immunoglobulin E (FcERII)]. The similar genomic organization of these three genes is discussed and consideration is given to the evolutionary duplications that may underlie this arrangement. Both DC-SIGN and DC-SIGNR possess a neck region, made up of multiple repeats, which supports the ligand-binding domain. Consideration is given to the biological reasons underlying the considerable polymorphism in the numbers of repeats in DC-SIGNR, but not DC-SIGN. The expression patterns of both DC-SIGN and DC-SIGNR are discussed in detail, with particular attention to the expression of both molecules in the placenta, which may have implications for the vertical transmission of HIV. Since dendritic cells may be important in determining the phenotype of many immune responses, via effects on T-lymphocytes, the differential expression of DC-SIGN by particular dendritic cell subsets may have important implications for the immunobiological functions of DC-SIGN. Similarly, the expression of DC-SIGNR by very restricted subsets of endothelial cells may give clues to the function of DC-SIGNR. Finally, the role of DC-SIGN in pathology, particularly in infective and neoplastic processes, is discussed, followed by speculation about likely future developments in this field. PMID- 12653691 TI - Dissociation of augmented physiological, hormonal and cognitive responses to hypoglycaemia with sustained caffeine use. AB - In patients with Type I diabetes and healthy volunteers, ingestion of modest amounts of caffeine augments the usual symptomatic and counter-regulatory responses to hypoglycaemia. The aim of the present study was to determine whether these are lost with sustained caffeine use, i.e. does tolerance develop? Eleven healthy caffeine consumers underwent two identical hyperinsulinaemic glucose clamp procedures. For 7 days prior to each clamp, subjects consumed a caffeine free diet supplemented with 200 mg of caffeine capsules twice daily (caffeine replete) or placebo (caffeine-withdrawn). During each clamp, blood glucose was held for 80 min at 4.5 mmol/l and then 2.5 mmol/l. At 85 min, subjects were given a 200 mg caffeine capsule. Measurements were taken of symptoms, plasma catecholamine, middle cerebral artery blood velocity (V(MCA)) and cognition. Following the acute caffeine challenge and during hypoglycaemia, V(MCA) fell only in the caffeine-withdrawn condition [-5.1 (-7.3, -3.0) cm/s compared with -1.9 ( 4.0, +0.2) cm/s in caffeine-replete condition; P <0.04; values are differences (95% confidence intervals)]. Plasma catecholamine levels and global cognitive performance were unaffected by caffeine status, whereas tests of executive intellectual function were better preserved during hypoglycaemia in the caffeine replete condition ( P <0.05). The influence of caffeine on hypoglycaemic symptomatic awareness depended upon the duration of the hypoglycaemic stimulus. At onset, symptoms were more intense in caffeine-withdrawn state ( P <0.01); however, with increasing duration of hypoglycaemia, symptom intensity was greater in caffeine-replete condition ( P <0.05). Thus previous caffeine consumption influences the physiological and symptomatic responses to acute hypoglycaemia, but complete tolerance does not develop with sustained use. PMID- 12653694 TI - Aspirin sensitivity and urticaria. AB - The relationship of aspirin sensitivity to urticaria is complex. Aspirin sensitivity can cause acute urticaria in some individuals, aggravate pre-existing chronic urticaria in others or, rarely, act as a cofactor with food or exercise to provoke anaphylaxis. Individuals who react with urticaria appear to come from a different population to those who react with asthma, although there is some overlap. Aspirin-sensitive chronic urticaria patients may also react adversely to some food additives. The pharmacological mechanisms of aspirin-sensitive urticaria are not fully understood but probably involve diversion of arachidonic acid metabolism from prostaglandin to cysteinyl leukotriene formation leading to direct effects on blood vessels and delayed mast cell degranulation with release of histamine. Cross-reactivity amongst all nonsteroidal drugs is common in aspirin-aggravated chronic urticaria but appears not to occur with selective cyclo-oxygenase 2 inhibitors. PMID- 12653696 TI - Larval therapy--an effective method of ulcer debridement. AB - Since ancient times, larval therapy has been reputed to help wound healing. Its use has recently been rediscovered and it is now enjoying increasing interest in clinical practice, as well as in research. We have investigated the effects of larval therapy on wounds in an open study of 74 patients with necrotic or sloughy chronic ulcers of different aetiologies. We found larval therapy to effectively debride 86% of the necrotic ulcers, and a single application was clinically beneficial in two-thirds of the patients. Failure to debride was mostly attributable to larval death. No ulcer type was shown to be more suited to larval therapy than others; however, there was an excellent response in all 29 patients with diabetes. Larval therapy was also noted to reduce odour in 58% of the 31 foul-smelling ulcers of mixed aetiology. No serious side-effects were observed. One-quarter of the study group experienced less pain during treatment, while 41% felt no difference in pain, and, although 34% noted an increase in pain, most of these patients wanted to continue the treatment because of subjective and objective visual improvement in wound debridement. In conclusion, we found larval therapy to be effective for debriding ulcers, easy to use and well accepted by the patients. PMID- 12653695 TI - Lichen sclerosus. AB - Lichen sclerosus is a chronic skin condition, which offers many challenges to the clinician. It affects men, women and children, and usually occurs in the anogenital area. The clinical signs can be confused with those seen in sexual abuse in children. The underlying cause is unknown; however, there is a strong association with autoimmune disorders, and immunogenetic studies have demonstrated a link with HLA DQ7. Patients suffer significant morbidity as a consequence of the intractable symptoms, physical scarring and psychosexual damage. Support groups may be helpful for some patients. Potent topical corticosteroids have been shown to be effective. There is a 5% incidence of squamous cell carcinoma, and all suspicious lesions should be biopsied. It is unclear whether the risk of malignancy is changed with the use of topical corticosteroids, as there is a potential risk of triggering a latent infection of human papillomavirus. A multidisciplinary approach to care is required and ideally all patients should attend a dedicated clinic and be offered long-term monitoring. PMID- 12653697 TI - Thirty-one cases of Marjolin's ulcer. AB - Marjolin's ulcer is a rare and often aggressive cutaneous malignancy that arises in previously traumatized or chronically inflamed skin, particularly after burns. We reviewed 264 burns cases treated in our departments to assess the frequency and clinicopathological features of this malignant complication. We found 31 cases of Marjolin's ulcer and a further 14 cases of nonmalignant ulceration at previous burn sites. Eighteen Marjolin's ulcers were located on the extremities, six on the scalp, six on the trunk, and one on the nose. Most malignancies were squamous cell carcinomas. The average time lag between the burns and subsequent malignant ulceration was 19 years. Four of the malignant ulcers had spread to regional lymph nodes, but there were no distant metastases. This study emphasizes that Marjolin's ulcer should be considered as a significant postburn complication. PMID- 12653698 TI - Kikuchi-Fujimoto disease with papulopustular skin manifestations. AB - Kikuchi-Fujimoto disease commonly presents with cervical lymphadenopathy accompanied by fever, myalgia, neutropaenia, and rarely cutaneous eruption. Most cutaneous lesions present as erythematous macules, papules, plaques, nodules, or ulcers on the upper part of the body such as trunk, upper extremities, and face. We present a case of Kikuchi-Fujimoto disease with a papulopustular eruption on the whole body including the lower extremities. PMID- 12653699 TI - Benign cutaneous Degos' disease. AB - Malignant atrophic papulosis is a rare systemic vaso-occlusive disorder characterized by thrombosis of vessels of the dermis, gastrointestinal tract, central nervous system and, occasionally, other organs. Cutaneous lesions consist of erythematous, dome-shaped papules that develop a central area of necrosis to leave a porcelain-like scar. The most accepted theory of pathogenesis is based on endothelial cell damage. There is no effective treatment of the disease. We describe a 26-year-old man with Degos' disease, a diagnosis based on the clinical and histologic pattern of skin lesions. The good response to antiplatelet therapy and the absence of systemic involvement over 8 years' follow-up is noteworthy. We believe that this case represents the benign form of the disease, typically referred to as benign cutaneous Degos' disease. PMID- 12653700 TI - Aleukaemic leukaemia cutis presenting as a benign-appearing eruption. AB - A 68-year-old Caucasian male presented with a 5-week history of a widespread pruritic papular eruption. Histology from a papule on the left shoulder showed a dense dermal infiltrate of large mononuclear cells which were positive for leucocyte common antigen, KP1 and PGM1, with an MIB-1 proliferating fraction of 40%, diagnostic of acute monocytic (M5) leukaemia cutis. Full blood count revealed pancytopaenia but no blasts. Bone marrow aspirate showed reduced red cell precursors and 10% blasts, consistent with myelodysplastic syndrome (refractory anaemia with excess blasts). The patient was managed with a 3 unit transfusion of packed red cells, after which his skin eruption resolved within 6 weeks and his peripheral blood counts returned to normal. No chemotherapy was administered. In conclusion, leukaemia can present in the skin, the eruption may be nonspecific and it may precede systemic involvement by either myelodysplastic syndrome or acute leukaemia. PMID- 12653702 TI - Leucocytoclastic vasculitis and influenza vaccination. AB - Influenza vaccination is recommended for all people over 75 years of age and for an expanding range of other indications. Side-effects of influenza vaccination are usually mild but we describe four cases of leucocytoclastic vasculitis following influenza vaccination. The four patients, who were all elderly, presented with cutaneous vasculitis but all had abnormal urinalysis suggestive of associated renal involvement. Since 1974 only 10 cases of vasculitis following influenza vaccination have been reported in the literature. The clinical features of these cases are reviewed. As the use of influenza vaccination is likely to increase, dermatologists should be aware of the possible association with vasculitis. PMID- 12653701 TI - Purpura and leg ulcers in a patient with cryoglobulinaemia, non-Hodgkin's lymphoma, and antiphospholipid syndrome. AB - We report a 69-year-old Caucasian female with non-Hodgkin's lymphoma who subsequently developed leg ulcers as a manifestation of the antiphospholipid syndrome. Investigations revealed a mixed cryoglobulinaemia with monoclonal IgM kappa and antiphospholipid activity with anticardiolipin antibodies, antimitochondrial type M5 antibodies and lupus anticoagulant. Significantly increased concentration of anticardiolipin antibodies was detected in the cryoprecipitate. Our case illustrates a connection between cryoglobulinaemia and lymphoproliferative and autoimmune disorders. Both cryoglobulins and anticardiolipin antibodies could participate in the vascular damage. Cutaneous manifestations in the presence of these disorders associated with non-Hodgkin's lymphoma have not been described previously. PMID- 12653703 TI - Mucor mycosis infection presenting as a non-healing ulcer in an immunocompromised patient. AB - Mucor mycosis is an uncommon saprophytic opportunistic fungus causing localized cutaneous infection associated with high morbidity and, on dissemination, high mortality. We report the case of an immunocompromised patient with an aggressively progressing, painful non-traumatic ulceration, unresponsive to standard treatment. Deep biopsies for haematoxylin and eosin staining and fungal culture revealed the characteristic broad non-septate irregular hyphae of mucor allowing introduction of the appropriate treatment. Infection with mucor mycosis must be considered in today's medical environment as the number of immunocompromised patients increases. PMID- 12653704 TI - Widespread morphoea following radiotherapy for carcinoma of the breast. AB - We report a case of a 60-year-old lady who was treated with radiotherapy for breast cancer of both breasts 8 years apart. Thirteen years after the first dose of radiotherapy she developed localized morphoea in all the irradiated skin of the chest wall and also the gaiter regions of both lower legs. Radiation-induced localized morphoea has been previously reported; however, there is no previous publication of an occurrence at a distant site as in this case. This case demonstrates that morphoea can occur distant to the original breast carcinoma and site of radiotherapy. We postulate that radiotherapy can induce neoantigen formation, which initiates a T cell response and subsequent tissue growth factor alpha release. Tissue growth factor alpha induces fibroblast activation and collagen production may persist due to a positive feedback mechanism within the fibroblast. The reason why the disease did not generalize remains unclear. PMID- 12653705 TI - Dystrophic epidermolysis bullosa complicated by cutaneous squamous cell carcinoma and pulmonary and renal amyloidosis. AB - A 25-year-old woman with Hallopeau-Siemens recessive dystrophic epidermolysis bullosa had generalized blistering, scarring and milia since birth. In the course of the disease, acral pseudosyndactyly developed, and the patient suffered from corneal erosions, oesophageal strictures, malabsorption, recurrent severe pneumonias and nephrotic syndrome. In addition, she had severe anaemia, sideropaenia, hypocalcaemia, heavy proteinuria and hypoalbuminaemia. A rapidly growing skin squamous cell carcinoma developed on the neck that spread to axillary and cervical lymph nodes. Recurrent hypocalcaemic tetanic convulsions and dyspnoea and a pneumonia refractory to antibiotics led to the premature demise of the patient. Autopsy revealed extensive amyloidosis of the renal, hepatic and splenic tissues. AA type amyloid deposits were detected in the renal glomeruli and in the lung, explaining the patient's unusually severe pulmonary infections. In essence, the patient had severe recessive dystrophic epidermolysis bullosa, complicated by squamous cell carcinoma, recurrent pneumonias and nephrotic syndrome due to secondary amyloidosis of the kidney and lung. The possibility of secondary pulmonary amyloidosis should be considered in severe dystrophic epidermolysis bullosa patients with recurrent pulmonary infections. PMID- 12653706 TI - Tea tree oil dermatitis associated with linear IgA disease. AB - Tea tree oil dermatitis is an increasingly common finding, reflecting the strong demand for natural remedies and aromatic substances. Linear immunoglobulin A (IgA) disease is a rare acquired subepidermal blistering disorder, characterized by basement membrane zone IgA deposition. We describe a patient in whom linear IgA disease appears to have been precipitated by a contact reaction to tea tree oil. PMID- 12653707 TI - Keloidal scleroderma. AB - A 53-year-old woman with systemic sclerosis developed numerous cutaneous nodules, resembling keloids, on her anterior chest wall. The nodules failed to respond to therapy with topical steroid, calcipotriol, or extended photochemotherapy. This is a rare, disfiguring variant of scleroderma, unresponsive to treatment, and poorly understood. PMID- 12653708 TI - Granulomatous mycosis fungoides presenting as an acquired ichthyosis. AB - We report a case of a 69-year-old gentleman who presented with a 3-month history of unexplained fevers and malaise who developed generalized pruritus, alopecia and an ichthyosiform erythematous eruption on his forearms, legs, chest and back. Skin histology, immunophenotyping and molecular features were consistent with granulomatous mycosis fungoides. He has been successfully treated with twice weekly PUVA photochemotherapy. PMID- 12653709 TI - Contact allergy: the role of skin chemistry and metabolism. AB - Chemical reactivity plays the driving role in the biological processes that result in the induction of allergic contact dermatitis. This paper presents an overview of the chemical basis of allergic contact dermatitis, including the physicochemical parameters governing skin penetration, chemical reaction mechanisms associated with haptenation of skin proteins, (quantitative) structure activity relationships (Q)SARs for contact allergens and prohaptens/skin metabolism of contact allergens. Despite the complexities and poor understanding of some of the metabolic processes leading to skin sensitization, it is possible to describe some of the relationships between chemical structures and the ability to form covalent conjugates with proteins. This knowledge, which relates chemical structure to a specific endpoint, can be programmed into an expert system. The Deductive Estimation of Risk from Existing Knowledge (DEREK) is one such expert system which is described in further detail. PMID- 12653710 TI - The central role of FcepsilonRI in allergy. AB - The high-affinity receptor for immunoglobulin E (IgE), FcepsilonRI, plays a central role in the initiation and control of atopic allergic inflammation. On mast cells and basophils, the function of the receptor is well known and constitutes cellular degranulation and the release of various mediators. FcepsilonRI on antigen-presenting cells (APCs), however, does not lead to degranulation of preformed granula, but has different functions: signal transduction pathways like the activation of NF-kappaB are initiated to induce inflammatory cytokine gene expression. In addition, FcepsilonRI on APCs acts as an allergen-focusing structure and can efficiently amplify allergen presentation in an IgE-dependent manner. Recently, we and others have gained new insight into the regulation and function of FcepsilonRI on APCs, which has shed new light on the modulating effects of the immune system in atopic inflammation. PMID- 12653711 TI - The role of free oxygen radicals in the aetiopathogenesis of rosacea. AB - A possible link between superoxide dismutase activity and malondialdehyde level with the clinical manifestations of rosacea was investigated. We found differences in superoxide dismutase activities between mild rosacea (stages I and II) and severe involvement (stage III) groups, as well as between disease and control groups that were statistically significant (P < 0.05). In the mild involvement group (stages I and II), the superoxide dismutase activity was higher than in the control group (P < 0.05), while the malondialdehyde levels did not differ from the control. In the severe involvement group (stage III), the superoxide dismutase activity was lower than in the control group (P < 0.05), and this was coupled to a raised level of malondialdehyde (P < 0.05). These findings clearly show that in the mild involvement phase of rosacea patients, superoxide dismutase activity was stimulated to protect the skin against reactive oxygen species so that the malondialdehyde levels were maintained. In contrast, in more severe disease, due to a decrease in the capacity of the antioxidant defence system, the malondialdehyde levels were increased. These findings support the 'antioxidant system defect hypothesis' in rosacea patients. PMID- 12653712 TI - An efficient gene transduction system for studying gene function in primary human dermal fibroblasts and epidermal keratinocytes. AB - One of the critical challenges for cellular genetic studies in primary human skin cells is lack of a gene delivery system that provides efficient transduction and sustained expression of the transgenes. Due to the limited time of survival in culture, the processes of drug selection and clonal expansion for establishing gene stably expressing cell lines are not a realistic option for primary skin cells. We have examined various gene transduction techniques in primary dermal fibroblasts and epidermal keratinocytes of human skin. We report here that vectors based on the human immunodeficiency virus (HIV, lentivirus) offer more than 90% gene transduction efficiency and sustained expression of transgenes in both human skin cell types. In contrast, most of the commonly used techniques have at best 30% transduction efficiency in these cells. Using two previously reported migration control genes, protein kinase Cdelta and p38alpha-MAPK, as examples, we provide evidence that the unprecedented efficiency of the lentiviral system enables a clear detection of the genes' dominant negative effects, which are otherwise greatly compromised by ordinary transfection techniques. We believe that a wide application of this gene transduction system will greatly benefit studies of gene function in human skin cells. PMID- 12653713 TI - A new extended haplotype Cw*0602-B57-DRB1*0701-DQA1*0201-DQB1*0201 associated with psoriasis in the Croatian population. AB - In this study, we have analysed the distribution of HLA class II alleles and the extended haplotype HLA-Cw-B-DRB1-DQA1-DQB1 in Croatian patients with type I and type II psoriasis by hybridization with specific oligonucleotide probes. Type I psoriasis showed a significant association with the DRB1*0701 [P < 0.00001; relative risk (RR) = 5.83], DQA1*0201 (P < 0.00001; RR = 6.12), DQB1*0201 (P = 0.0006; RR = 3.29) and DQB1*0303 alleles (P = 0.0008; RR = 7.51). A negative correlation with type I disease was observed for the DQA1*0102 allele (P = 0.002; RR = 0.26). Type II psoriasis did not show any association with any class II alleles. The extended haplotype HLA-Cw*0602-B57-DRB1*0701-DQA1*0201-DQB1*0201 was present at a significantly higher frequency in type I patients (P < 0.00001; RR = 7.72). However, this haplotype was not detected at all in patients with type II psoriasis. In conclusion, the extended haplotype HLA-Cw*0602-B57-DRB1*0701 DQA1*0201-DQB1*0201 is a risk haplotype for type I disease in the Croatian population. This particular haplotype has not been reported previously in association with psoriasis in any other ethnic groups. PMID- 12653714 TI - Fish odour syndrome with features of both primary and secondary trimethylaminuria. AB - We report a patient with the fish odour syndrome who has both primary and secondary trimethylaminuria. The diagnosis was made using biochemical and genetic analysis in the apparent absence of any characteristic smell. Differentiation of primary and secondary trimethylaminuria is usually made on urinary analysis of trimethylamine and its metabolite trimethylamine N-oxide, with different, characteristic patterns of both compounds in primary and secondary trimethylaminuria. Our patient had biochemical analysis consistent with a diagnosis of secondary trimethylaminuria, while analysis of the flavin-containing mono-oxygenase 3 gene, the causative gene in primary trimethylaminuria, demonstrated three sequence polymorphisms, two of which are known to reduce enzyme activity. The patient showed temporary clinical and biochemical response to treatment with metronidazole and neomycin. It is important to be aware of this diagnosis in patients without obvious clinical signs, and of the subjective benefits of treatment. PMID- 12653715 TI - Recurrent missense mutations in the hair keratin gene hHb6 in monilethrix. AB - Monilethrix is an autosomal dominant hair disorder characterized by a beaded appearance of the hair resulting from periodic thinning of the shaft (MIM 158000). The phenotype shows variable penetrance and results in hair fragility and patchy dystrophic alopecia. Mutations of the helix-encoded region in two hair specific keratins (hHb1 and hHb6) have been identified as responsible for this disorder. We investigated two unrelated families from Russia and Colombia with monilethrix and found two missense mutations in hHb6. In the Russian family, we found a G to A transition at the first base of codon 402, resulting in a lysine substitution (GAG to AAG), designated E402K. In the Colombian family, affected patients carried a missense mutation of codon 413, involving a transition from G to A causing a lysine substitution (GAG to AAG), designated E413K. These two mutations have been identified in other monilethrix families from Europe. Our findings extend the body of evidence implicating recurrent hHb6 and hHb1 mutations in monilethrix families from around the world. PMID- 12653716 TI - A novel tumour marker RCAS1 in a case of extramammary Paget's disease. AB - A 66-year-old male, whose primary skin lesion in extramammary Paget's disease had been surgically resected 4 years previously, was hospitalized with liver metastases. Hepatic arterial infusion chemotherapy was carried out and the tumours clearly reduced in size. Serum levels of some common tumour markers were not elevated, even prior to therapy. We measured serum levels of a novel tumour associated antigen, RCAS1, because its expression was detected in the tumour cells. The patient's serum RCAS1 level was elevated (22.0 U/mL) before therapy and fell during (10.5 U/mL) and after (5.0 U/mL) therapy. Therefore, serum RCAS1 levels may be valuable as a potential biomarker for monitoring therapeutic efficacy against Paget's disease. PMID- 12653717 TI - Impact of skin cancer education on medical students' diagnostic skills. AB - Skin cancer is increasingly common, and the skills involved in its diagnosis should be promoted in UK medical schools. However, there has been no scientific evaluation of the teaching methods employed by dermatology departments. The aim of this study was to evaluate, using traditional audiovisual methods, the impact of an illustrated booklet on skin cancer, coupled with a lecture, on undergraduates' diagnostic skills. The ability of 27 final-year medical students to recognize a variety of skin lesions, using projected images from clinical slides, was assessed. They were tested without warning on two occasions. Immediately after the first test, students were given an illustrated booklet on skin tumours and pigmented lesions which was supplemented with a lecture based on the booklet. Two weeks later, a second test was employed using a series of slides deemed to be of equal diagnostic difficulty. Our results showed a significant increase in the median number of correct diagnoses between the first and second tests (P < 0.001). However, there remained wide variation at the second test in the percentage of correct answers (30 to 80%) amongst students. Our study highlights the need to develop effective methods for improving the diagnostic skills of undergraduates in dermatology, and the importance of evaluating teaching methods. The methods of evaluation, such as ours, can be simple and inexpensive. PMID- 12653718 TI - Contact allergy: the local lymph node assay for the prediction of hazard and risk. AB - The prospective identification of potential contact allergens and their subsequent safety assessment are pivotal in successful management of this risk to human health. Although much can be learned from the chemical and physical properties of a substance, the definitive information in respect of sensitizing hazard/risk derives from an assessment of the integrated response of the immune system. In recent years, the focus for such assessments has begun to switch from the guinea pig to the mouse, notably to the local lymph node assay (LLNA). In this paper, the current value of the LLNA for hazard identification is reviewed and its regulatory status defined. Once a potential contact allergen has been identified, however, the vital clue to accurate safety evaluation is the assessment of the potency of the allergen. How this can be achieved using the LLNA and employed in safety evaluation is discussed in detail, together with practical suggestions for the deployment of such processes in regulatory toxicology. PMID- 12653719 TI - Solar urticaria induced by infrared radiation. PMID- 12653721 TI - Lack of response of elephantiasic pretibial myxoedema to treatment with high-dose intravenous immunoglobulins. PMID- 12653720 TI - Peristomal lichen sclerosus developing posturostomy. PMID- 12653722 TI - Scrotal metastases originating from colorectal carcinoma. PMID- 12653723 TI - Subungual haemorrhage in hydroa vacciniforme. PMID- 12653724 TI - Genital swelling caused by octyldodecanol contact dermatitis. PMID- 12653725 TI - Cutaneous reaction to oral antihistamine. PMID- 12653726 TI - Acquired port-wine stain associated with glaucoma. PMID- 12653728 TI - New CD56 positive and cytotoxic T-cell cutaneous lymphomas from the World Health Organisation. PMID- 12653729 TI - Causes, investigation and treatment of leg ulceration. AB - Chronic ulceration of the lower leg is a frequent condition, with a prevalence of 3-5% in the population over 65 years of age. The incidence of ulceration is rising as a result of the ageing population and increased risk factors for atherosclerotic occlusion such as smoking, obesity and diabetes. Ulcers can be defined as wounds with a 'full thickness depth' and a 'slow healing tendency'. In general, the slow healing tendency is not simply explained by depth and size, but caused by an underlying pathogenetic factor that needs to be removed to induce healing. The main causes are venous valve insufficiency, lower extremity arterial disease and diabetes. Less frequent conditions are infection, vasculitis, skin malignancies and ulcerating skin diseases such as pyoderma gangrenosum. But even rarer conditions exist, such as the recently discovered combination of vasculitis and hypercoagulability. For a proper treatment of patients with leg ulcers, it is important to be aware of the large differential diagnosis of leg ulceration. PMID- 12653730 TI - Guidelines for treatment of onychomycosis. AB - These guidelines for management of onychomycosis have been prepared for dermatologists on behalf of the British Association of Dermatologists. They present evidence-based guidance for treatment, with identification of the strength of evidence available at the time of preparation of the guidelines, and a brief overview of epidemiological aspects, diagnosis and investigation. PMID- 12653731 TI - Decreased cutaneous expression of stem cell factor and of the p75NGF receptor in urticaria. AB - BACKGROUND: Mast cells, the main effector cells in urticaria, have been reported to be increased in number in lesional and nonlesional skin of urticaria patients, but the underlying mechanisms have so far not been studied. Serum NGF has however, been reported to be increased in urticaria. OBJECTIVES: We have therefore explored the potential involvement of known mast cell growth modulating factors in urticaria. METHODS: Tissue sections from patients with different types of urticaria and healthy controls were studied for the immunohistochemical expression of known mast cell growth factors (stem cell factor, SCF; nerve growth factor, NGF), of the inhibitory granulocyte-macrophage colony-stimulating factor (GM-CSF), and of the corresponding receptors, using the alkaline phosphatase antialkaline phosphatase technique. RESULTS: Compared to skin of normal controls, staining for SCF, but not for NGF and GM-CSF, was significantly decreased in epidermis, endothelium and perivascular cells in lesional and nonlesional skin of all urticarias. On separate analysis of urticaria subtypes, decreased expression reached significance only in delayed pressure urticaria. Expression of the p75NGF receptor (p75NGFR) was also significantly decreased on endothelium and on perivascular cells of lesional and nonlesional skin in all urticarias. On evaluation of serial sections, p75NGFR expression was also decreased on c-Kit positive dermal mast cells. In contrast, expression of the NGF receptor tyrosine kinase and of the SCF and GM-CSF receptors was unchanged. CONCLUSIONS: These findings show that SCF and p75NGFR are selectively and systemically down regulated in the skin of urticaria patients and may represent a negative feedback to increased mast cell reactivity and proliferation. PMID- 12653732 TI - A study of candidate genes for psoriasis near HLA-C in Chinese patients with psoriasis. AB - BACKGROUND: Genetic analyses have identified the HLA-Cw6 allele as the major risk allele for psoriasis in many racial groups. However, by serological typing, HLA Cw6 is not considered a risk factor in Chinese psoriatics. There are several susceptibility genes for psoriasis residing in chromosome 6p near the HLA-C locus, including the corneodesmosin (CDSN) gene, the octamer transcription factor 3 (POU5F1) gene, the major histocompatibility complex class I chain-related gene A (MICA), and the gene for tumour necrosis factor (TNF)-alpha. However, the information about their role in psoriasis in Chinese patients is limited. OBJECTIVES: We aimed to determine whether Cw6 and the genetic polymorphism of the CDSN gene, POU5F1 gene, MICA gene and the gene for TNF-alpha promoter region were associated with an increased risk of psoriasis in Chinese patients. METHODS: We conducted a case-control association study in 105 Chinese patients with psoriasis vulgaris and 160 control subjects of similar ages. Genotypes of Cw6, the CDSN gene, the POU5F1 gene, and the gene for the TNF-alpha promoter region were determined by polymerase chain reaction (PCR) followed by restriction enzyme digestion. Genotyping of MICA was determined by PCR combined with fluorescent based automated fragment detection technology. Results The allele frequencies showed no differences between patients and controls for the POU5F1 gene, MICA gene and the gene for TNF-alpha promoter region. The frequency of the HLA-Cw6 allele in the psoriasis group was significantly higher than that in the control group (18.6% vs. 6.56%, P < 0.00005). For the CDSN gene, patients were more likely to have C allele at position +619 (P = 0.006) and C allele at position +1243 (P = 0.007), but the significance disappeared after correction for multiple testing (Pc > 0.05). CONCLUSIONS: HLA-Cw6 remains the most significant susceptibility gene in Chinese patients with psoriasis. However, the role of the CDSN gene in the pathogenesis of psoriasis deserves further scrutiny. PMID- 12653733 TI - Early vitronectin receptor downregulation in a melanoma cell line during all trans retinoic acid-induced apoptosis. AB - BACKGROUND: Recent evidence assigns the vitronectin receptors (VnRs) an important role in regulating tumour cell invasion and dissemination. In vivo and in vitro studies document that all trans-retinoid acids (ATRAs) inhibit growth-inducing apoptosis in melanomas. OBJECTIVES: We have analysed the effects of ATRA treatment on melanoma cell adhesion and motility. METHODS: Human M14 melanoma cells were treated with 10 micromol L-1 ATRA for different times and stained with rhodamine-phalloidin to analyse the effect of treatment on cytoskeleton organization. Cell adhesion and cell migration assays were performed to analyse the role of VnRs in the ATRA-induced early stages of apoptosis. VnR expression was evaluated by Western blot, immunoprecipitation and immunocytochemistry assays. RESULTS: First, using an annexin V assay, we found that apoptosis was triggered by 48 h with 10 micromol L-1 ATRA exposure. At this time point, decrease in the F-actin polymerization as well as inhibition of cell adhesive ability to vitronectin (Vn) was exerted by ATRA treatment. In the presence of serum, exposure to 10 micromol L-1 ATRA for 48 h produced a dramatic inhibition of the cell adhesion ability that was comparable with that exerted by untreated cells preincubated with anti-alpha(v)beta(3) or anti-alpha(v)beta(5) VnR monoclonal antibodies. Functionally, the treatment of melanoma cells with 10 micromol L-1 ATRA for 48 h causes an inhibition of directional cell migration towards Vn-coated filters. Therefore, we analysed the effect of ATRA on the VnR expression. Both alpha(v)beta(3) and alpha(v)beta(5) VnR levels were reduced upon exposure to 10 micromol L-1 ATRA for 48 h as shown by Western blot, immunoprecipitation and immunocytochemistry assays. CONCLUSIONS: Altogether, our data indicate that treatment of M14 melanoma cells with ATRA downregulates VnR expression and that this reduction is closely correlated with the ATRA-dependent inhibition of actin-fibre organization, cell adhesion and migration. Although the mechanism by which ATRA regulates the expression of VnR in M14 melanoma cells needs further elucidation, this system may represent a model for understanding the molecular basis of ATRA therapy in melanoma. PMID- 12653734 TI - The role of stratum corneum and dermal microvascular perfusion in penetration and tissue levels of water-soluble drugs investigated by microdialysis. AB - BACKGROUND: Hydrophilic drugs are poorly absorbed when applied topically, due to low partitioning through the lipid matrix of the stratum corneum. Cutaneous blood flow rapidly clears the absorbed drug, which may result in low tissue levels. This is of importance for topically applied drugs whose site of action is within the epidermis or dermis. Dermal drug levels can be measured using cutaneous microdialysis, which is a means of continuously sampling substances from the dermal extracellular fluid. OBJECTIVES: To measure the contribution of stratum corneum barrier and microvascular perfusion in determining dermal tissue levels of hydrophilic drugs (aciclovir and penciclovir) in vivo. METHODS: Studies were performed using microdialysis of the volar surface of the forearm of healthy volunteers (n = 55) over a 5-h collection period. Stratum corneum was removed by tape stripping, and barrier disruption quantified by measurement of transepidermal water loss (TEWL); dermal microvascular perfusion was modulated by inclusion of noradrenaline in the microdialysis perfusate. RESULTS: With intact skin and normal cutaneous blood flow the concentration of penciclovir recovered was below assay threshold (0.05 ng x mL(-1). With noradrenaline-induced local vasoconstriction, the area under the curve of drug absorbed through normal skin (+/- SEM) was 13.3 +/- 2.9 ng mL(-1) h(0-5) for penciclovir and 27.6 +/- 10.6 ng mL(-1) h(0-5) for aciclovir. Removal of the stratum corneum (to glistening) by tape stripping increased penciclovir absorption by 1300-fold and aciclovir absorption by 440-fold, confirming the stratum corneum as the major barrier to hydrophilic drug absorption. Sequential barrier disruption by tape stripping gave a close correlation between penciclovir concentration absorbed per hour and barrier disruption measured by TEWL (r2 = 0.9283). There was a 15.6-fold difference in the recovery of penciclovir through barrier-deficient skin with and without cutaneous blood flow. There was no relationship between fibre depth and amount of drug dialysed, which suggests free movement of antiviral drug on reaching the aqueous environment of the dermis. CONCLUSIONS: This study defines for the first time the relationship between the degree of mechanical barrier impairment and drug absorption at the same anatomical site in humans, and the role of blood flow in drug clearance in vivo. PMID- 12653735 TI - Beneficial effects of fumarate therapy in psoriasis vulgaris patients coincide with downregulation of type 1 cytokines. AB - BACKGROUND: Fumarates have been shown to be effective in psoriasis vulgaris. OBJECTIVES: To find out whether successful therapy is associated with modulation of cytokines. METHODS: We determined interferon (IFN)-gamma, interleukin (IL)-4 and IL-10 secretion capacities of peripheral blood mononuclear cells (PBMC) after phytohaemagglutinin stimulation, and IL-12p70 and IL-10 secretion capacities of PBMC after endotoxin stimulation in psoriasis vulgaris patients during treatment with fumarates. In a cohort study, 12 patients (five men, median age 50 years; seven women, median age 46 years) with psoriasis vulgaris were followed during 24 months of fumarate treatment. In addition, we followed 14 healthy controls (six men, median age 31 years; eight women, median age 29 years) without skin diseases during 12 months to investigate possible changes in the cytokine secretion capacity of PBMC as a result of seasonal changes. Disease activity in patients was determined by Psoriasis Area and Severity Index (PASI) score. Blood was collected for measurement by enzyme-linked immunosorbent assay of cytokine levels after stimulation of PBMC. RESULTS: Within 6 months of fumarate treatment, the mean +/- SD PASI score had decreased to 22 +/- 9% of its initial value. These beneficial effects coincided with lymphocytopenia and a significant (P < 0.05) downregulation of IFN-gamma expression by circulating blood cells, followed by a significant downregulation of IL-4 expression. Notably, production of the cytokine synthesis inhibitor IL-10 by PBMC was unchanged. CONCLUSIONS: The beneficial effects of fumarates may be attributed to their downregulatory action on type 1 cytokines. PMID- 12653736 TI - Keratin 17 mutation in pachyonychia congenita type 2 with early onset sebaceous cysts. AB - BACKGROUND: Pachyonychia congenita (PC) is a group of autosomal dominant ectodermal dysplasias caused by mutations in four differentiation-specific keratin genes. Two major clinical subtypes of PC have been generally recognized. Symmetrically thickened fingernails and toenails are the defining characteristic of PC type 2 (PC-2) with onset at infancy. Pilosebaceous cysts are the best hallmark of PC-2, but they usually occur at puberty. OBJECTIVES: To report a Chinese pedigree of PC-2 with unusually early onset sebaceous cysts and to explore the genetic mutation and its phenotype. METHODS: Exon 1 of keratin 17 was amplified by polymerase chain reaction (PCR) from genomic DNA from the three patients in the pedigree, the proband, his half-sister and his younger son, two unaffected members in the pedigree and 50 unrelated and unaffected people. PCR products were directly sequenced to detect the mutation. RESULTS: Direct sequencing of the PCR products revealed a heterozygous 275A-->G mutation in all three affected members. This mutation predicts the substitution of asparagine by serine in codon 92 (N92S) located in the 1A domain of keratin 17. CONCLUSIONS: Mutation in the 1A domain of keratin 17 underlies the affected members' phenotype, PC-2 with early onset sebaceous cysts and late-onset thickened fingernails and toenails. The onset of the cysts is very early in some people within this family and the age at onset of thickened fingernails and toenails is variable within the family, implying the existence of modifying factors. PMID- 12653737 TI - Anaerobic cocci populating the deep tissues of chronic wounds impair cellular wound healing responses in vitro. AB - BACKGROUND: Anaerobic cocci are estimated to be present in the deep tissues of over 50% of chronic skin wounds. While the part they play in the chronicity of these wounds is uninvestigated, anaerobic cocci have previously been shown to be involved in other chronic inflammatory human conditions. METHODS: In this study the anaerobic microflora of the deep tissues of 18 patients with refractory chronic venous leg ulcers (mean age 80.3 years; mean duration > 24 months) was characterized using strict anaerobic culture conditions. The effect of the anaerobic organisms isolated from these tissues on extracellular matrix (ECM) proteolysis and cellular wound healing responses was studied using in vitro models. RESULTS: Anaerobic organisms were present in the deep tissues of 14 of 18 wounds and were principally Peptostreptococcus spp. The effects of three Peptostreptococcus spp. isolated from these wounds (P. magnus, P. vaginalis and P. asaccharolyticus) on cellular wound healing responses were compared with those of two pathogenic organisms also isolated from these wounds (Pseudomonas aeruginosa and Citrobacter diversus). While the direct ECM proteolytic activity exhibited by the Peptostreptococcus spp. was limited, they did significantly inhibit both fibroblast and keratinocyte proliferation, but only at high concentrations. However, at lower concentrations peptostreptococcal supernatants profoundly inhibited keratinocyte wound repopulation and endothelial tubule formation. The magnitude of these effects varied between strains and they were distinct from those demonstrated by Pseudomonas aeruginosa and Citrobacter diversus. CONCLUSIONS: These studies confirm the importance of anaerobic organisms in chronic wounds and demonstrate an indirect, strain-specific mechanism by which these microorganisms may play a part in mediating the chronicity of these wounds. PMID- 12653738 TI - Antibiotic-resistant acne: lessons from Europe. AB - BACKGROUND: Propionibacterium acnes and P. granulosum are widely regarded as the aetiological agents of inflammatory acne. Their proliferation and metabolism are controlled using lengthy courses of oral and/or topical antibiotics. Despite numerous reports of skin colonization by antibiotic-resistant propionibacteria among acne patients, accurate prevalence data are available only for the U.K. OBJECTIVES: To determine the prevalence of skin colonization by antibiotic resistant propionibacteria among acne patients and their contacts from six European centres. METHODS: Skin swabs were collected from 664 acne patients attending centres in the U.K., Spain, Italy, Greece, Sweden and Hungary. Phenotypes of antibiotic-resistant propionibacteria were determined by measuring the minimum inhibitory concentrations (MIC) of a panel of tetracycline and macrolide, lincosamide and streptogramin B (MLS) antibiotics. Resistance determinants were characterized by polymerase chain reaction (PCR) using primers specific for rRNA genes and erm(X), followed by nucleotide sequencing of the amplified DNA. RESULTS: Viable propionibacteria were recovered from 622 patients. A total of 515 representative antibiotic-resistant isolates and 71 susceptible isolates to act as control strains were characterized phenotypically. The prevalence of carriage of isolates resistant to at least one antibiotic was lowest in Hungary (51%) and highest in Spain (94%). Combined resistance to clindamycin and erythromycin was much more common (highest prevalence 91% in Spain) than resistance to the tetracyclines (highest prevalence 26.4% in the U.K.). No isolates resistant to tetracycline were detected in Italy, or in Hungary. Overall, there were strong correlations with prescribing patterns. Prevalence of resistant propionibacteria on the skin of untreated contacts of the patients varied from 41% in Hungary to 86% in Spain. Of the dermatologists, 25 of 39 were colonized with resistant propionibacteria, including all those who specialized in treating acne. None of 27 physicians working in other outpatient departments harboured resistant propionibacteria. CONCLUSIONS: The widespread use of topical formulations of erythromycin and clindamycin to treat acne has resulted in significant dissemination of cross-resistant strains of propionibacteria. Resistance rates to the orally administered tetracycline group of antibiotics were low, except in Sweden and the U.K. Resistant genotypes originally identified in the U.K. are distributed widely throughout Europe. Antibiotic-resistant propionibacteria should be considered transmissible between acne-prone individuals, and dermatologists should use stricter cross-infection control measures when assessing acne in the clinic. PMID- 12653739 TI - Atopy patch test reactions to Malassezia allergens differentiate subgroups of atopic dermatitis patients. AB - BACKGROUND: The yeast Malassezia is considered to be one of the factors that can contribute to atopic dermatitis (AD). OBJECTIVES: To investigate the reactivity to Malassezia allergens, measured as specific serum IgE, positive skin prick test and positive atopy patch test (APT), in adult patients with AD. METHODS: In total, 132 adult patients with AD, 14 with seborrhoeic dermatitis (SD) and 33 healthy controls were investigated for their reactions to M. sympodialis extract and three recombinant Malassezia allergens (rMal s 1, rMal s 5 and rMal s 6). RESULTS: Sixty-seven per cent of the AD patients, but only one of the SD patients and none of the healthy controls, showed a positive reaction to at least one of the Malassezia allergens (extract and/or recombinant allergens) in at least one of the tests. The levels of M. sympodialis-specific IgE in serum correlated with the total serum IgE levels. Elevated serum levels of M. sympodialis-specific IgE were found in 55% and positive APT reactions in 41% of the AD patients with head and neck dermatitis. A relatively high proportion of patients without head and neck dermatitis and patients with low total serum IgE levels had a positive APT for M. sympodialis, despite lower proportions of individuals with M. sympodialis specific IgE among these groups of patients. CONCLUSIONS: These results support that Malassezia can play a role in eliciting and maintaining eczema in patients with AD. The addition of an APT to the test battery used in this study reveals a previously overlooked impact of Malassezia hypersensitivity in certain subgroups of AD patients. PMID- 12653740 TI - Erosive pustular dermatosis of the leg associated with compression bandaging and fungal infection. AB - BACKGROUND: Erosive pustular dermatosis of the leg (EPDL) has been described in association with venous insufficiency and atrophy of the skin of the lower leg. Like erosive pustular dermatosis of the scalp, this disease has also been reported to be a non-infective condition. OBJECTIVES: To investigate the clinicopathological features and, where possible, the aetiology of clinical EPDL. METHODS: We identified a group of patients undergoing continuous compression bandaging for venous dermatitis of the legs and/or predominantly venous leg ulceration with clinical features described in patients with EPDL. They were investigated by skin biopsy, patch testing and microbiological tests for the presence of bacteria and fungi. RESULTS: Twenty-four of 400 (6%) patients were noted to have pustules on the leg(s). There was laboratory evidence of fungal infection in 13 of 24 patients (54%), with complete and sustained resolution of pustules after antifungal treatment. Pustulation in the other 11 patients (46%) was unresponsive to antibiotics for confirmed bacterial infection; some improvement was seen with potent topical corticosteroids but full clearance was achieved only after a switch from continuous four-layer compression bandaging to the use of intermittent long stretch compression. CONCLUSIONS: EPDL is a fairly common clinical picture seen in patients undergoing continuous compression bandaging. It may be produced by opportunistic, particularly fungal, infection. In almost half an infective aetiology cannot be demonstrated and a pyoderma gangrenosum-like process may be implicated. PMID- 12653741 TI - Skin diseases associated with the cosmetic use of bleaching products in women from Dakar, Senegal. AB - BACKGROUND: The cosmetic use of bleaching products is considered a common practice in dark-skinned women from sub-Saharan Africa. However, there are few studies on this subject. OBJECTIVES: To increase the knowledge about the dermatological consequences of this practice in Dakar, the capital of Senegal. METHODS: A representative sample of 368 adult women presenting at our dermatological centre was selected. Each woman was questioned about her cosmetic use of bleaching products. Next, the following data were recorded in 425 women who used bleaching products: names and types of products used; modalities of the skin bleaching practice; skin diseases motivating the dermatological visit, with recording of their clinical features; and results of a full skin examination. The active substances of the bleaching products were determined mainly by reading the indications on their packages; with products of unknown composition, a pharmacological analysis of samples was done. A statistical analysis was performed. RESULTS: Of the 368 women questioned, 194 (52.7%) were current users of bleaching products. Concerning the 425 users enrolled, products were applied on the whole body in 92% of users, with a median duration of use of 4 years. The active principles used included hydroquinone (used by 89% of users), glucocorticoids (70%), mercury iodide (10%) and caustic agents (17%); 13% of users used products of unknown composition. In the samples that were analysed, hydroquinone was found at concentrations of between 4% and 8.7%. Concerning steroids, superpotent (class 1) glucocorticoids predominated. The main skin complaints in bleaching products users included dermatophyte infections (n = 105) and scabies (n = 69), both often unusually extensive and severe; acne (n = 42), often severe; eczema (n = 41); irritant dermatitis (n = 14); and dyschromia (n = 26, including 14 cases of exogenous ochronosis). The skin examination noted features apparently disregarded by users: striae (noticed in 39% of users), and macular hyperchromia involving the face, mainly the periocular area (33%). The statistical analysis showed that glucocorticoid use was associated with the presence and severity of infectious skin diseases, and of acne. CONCLUSIONS: More than half of the adult women presenting at our dermatology centre were using bleaching products. Most skin diseases observed in bleaching products users appeared to be induced, aggravated or modified by this practice. Superpotent topical glucocorticoids appeared to be the main agents responsible for the observed complications. The cosmetic use of bleaching products therefore has a major impact on our current dermatological practice. PMID- 12653742 TI - Clinical and aetiological aspects in urticaria and angio-oedema. AB - BACKGROUND: Urticaria is a common disorder that affects as many as 20% of all people at some time during their lifetime. OBJECTIVES: To analyse the prevalence of various forms of urticaria according to an aetiological and clinical classification, we carried out a 4-year study in an outpatient clinic. METHODS: The study was carried out on 562 consecutive patients (178 males and 384 females; mean age 35.4 +/- 16), who had been referred to our unit for the study of urticaria and angio-oedema. Baseline investigations included: the patient's family and personal history of allergy; duration of symptoms, presence of associated symptoms and objective signs of the current episode; clinical, laboratory and instrumental investigations; assessment of response to antihistamine treatment. RESULTS: A family history of atopy was present in 35% of patients and a personal history of allergy in 24%. We subdivided urticaria and angio-oedema into several groups on the basis of their clinical and aetiological aspects. Of the 562 patients, 424 (76%) presented with ordinary urticaria (43 acute urticaria, 311 chronic urticaria, 70 episodic urticaria), 80 (14%) physical urticaria, 49 (9%) angio-oedema without weals, six (1%) IgE-mediated contact urticaria and three (0.5%) urticarial vasculitis. In 64 cases (11%) urticaria/angio-oedema was associated with one or more symptoms. We identified 394 cases (82%) of idiopathic urticaria, 42 (9%) of immunological urticaria, 29 (6%) of non-immunological urticaria and 17 (3%) of urticaria secondary to infections. Of the treated subjects, 54% showed a good response to treatment with antihistamines. CONCLUSIONS: Our data provide an overview of urticaria/angio oedema in a large series of patients, based on clinical-aetiological aspects, and related to recent diagnostic guidelines. PMID- 12653743 TI - Blastic natural killer cell and extranodal natural killer cell-like T-cell lymphoma presenting in the skin: report of six cases from the UK. AB - BACKGROUND: Some lymphomas express natural killer (NK)-cell markers such as the neural cell adhesion molecule, which is recognized by the CD56 antibody. These lymphomas may present in the skin, but do not represent a homogeneous group. The new World Health Organization classification of lymphoma/leukaemia recognizes several types of NK/T-cell neoplasm, including blastic NK-cell lymphoma, which characteristically presents with cutaneous lesions. OBJECTIVES: To describe the clinical, pathological and molecular features in six cases of CD56+ lymphoma with cutaneous presentation. METHODS: The clinical, histopathological and immunophenotypic features of six patients were reviewed. In addition, in situ hybridization (ISH) to identify Epstein-Barr virus (EBV) mRNA, and polymerase chain reaction analysis to identify the presence of a clonal population of T cells or B cells were performed on lesional skin. RESULTS: All patients presented with widespread nodules and plaques, which in five cases were a characteristic purple colour. Four patients developed disseminated disease, three with neurological involvement. These four patients died between 14 and 46 months following diagnosis (median 30 months). In four of six cases the histopathological and immunohistological features were in keeping with a blastic NK-cell lymphoma. No clonal immunoglobulin heavy chain (IgH) or T-cell receptor (TCR) gene rearrangement was detected in the four cases consistent with an origin from NK cells. A further case fitted the criteria for an extranodal NK/T-cell lymphoma of nasal type and was also the only case to show evidence of EBV mRNA by ISH. A clonal T-cell population was identified in the final case. This patient also exhibited molecular evidence of a clonal B-cell population and a t(14;18) translocation confirmed by sequence analysis. CONCLUSIONS: Our data confirm that NK-cell lymphomas presenting in the skin are a heterogeneous group, and that in the U.K., blastic NK-cell lymphoma is more common than extranodal NK/T-cell lymphoma of nasal type. These lymphomas pursue an aggressive course, with rapid development of disseminated disease, and resistance to chemotherapy. Detailed immunophenotyping is needed to distinguish the different types. Our molecular data indicate that blastic NK-cell lymphoma cases lack clonal TCR/IgH gene rearrangements consistent with an NK-cell origin. Our ISH findings indicate that EBV plays a pathogenetic role only in extranodal NK/T-cell lymphoma of nasal type. PMID- 12653744 TI - Subcutaneous panniculitis-like T-cell lymphoma: a clinicopathological, immunophenotypic and molecular analysis of six patients. AB - BACKGROUND: Subcutaneous panniculitis-like T-cell lymphoma (SPTCL) is a rare cytotoxic T-cell lymphoma of the skin. In the World Health Organization classification of T-cell and natural killer cell lymphoma it is listed as an example of extranodal lymphoma. In practice, however, it is most likely to present to a dermatologist. OBJECTIVES: To describe the clinicopathological, immunophenotypic and molecular features of six U.K. patients with SPTCL. METHODS: The clinical, histological and immunophenotypic features were reviewed. T-cell receptor (TCR) gene analysis was performed on blood and tissue samples using polymerase chain reaction/single-strand conformational polymorphism analysis of the TCR-gamma gene using consensus primers. In situ hybridization was performed on lesional skin to detect mRNA for Epstein-Barr virus (EBV). RESULTS: All patients presented with subcutaneous nodules, plaques or ulceration, and three had systemic symptoms. All biopsies exhibited an infiltrate of medium to large pleomorphic cells involving the subcutis with characteristic rimming of fat spaces. Five showed areas of necrosis, but only one showed marked cytophagia. In three cases the neoplastic cells did not express TCR-beta. One was strongly p53 positive, and the other two were CD56 positive. Both these patients showed epidermal involvement with lichenoid changes histologically, and both developed the haemophagocytic syndrome. The other three cases were TCR-beta positive, CD8 positive and CD56 negative. All cases were positive with pan T-cell markers and also for the cytotoxic granule protein T-cell intracellular antigen-1 and granzyme B. All cases were EBV negative both by immunostaining (latent membrane protein-1) and by in situ hybridization (EBV-encoded mRNA). TCR gene analysis revealed a T-cell clone in four of five cases; two of these patients had an identical T-cell clone in the peripheral blood. The median survival was 16 months. However, two of the three TCR-beta-negative patients have died, whereas none of the TCR-beta-positive patients has died. CONCLUSIONS: This is the first series of SPTCL patients to be reported in the U.K. and the data support the view that there are two subsets of SPTCL: those derived from gammadelta T cells which carry a poor prognosis, and are usually CD56 positive, and a more indolent group derived from alphabeta T cells. PMID- 12653745 TI - Confocal laser scanning microscopic observation of glycocalyx production by Staphylococcus aureus in skin lesions of bullous impetigo, atopic dermatitis and pemphigus foliaceus. AB - BACKGROUND: Glycocalyx collapses during dehydration to produce electron-dense accretions. Confocal laser scanning microscopy (CLSM) may be used to visualize fully hydrated microbial biofilms. OBJECTIVES: Using CLSM, to analyse glycocalyx production by Staphylococcus aureus cells in skin lesions of bullous impetigo, atopic dermatitis and pemphigus foliaceus. A second objective was to compare numbers of S. aureus cells in tissue sections prepared by different methods for routine light microscopy. METHODS: S. aureus cells in skin lesions of impetigo, atopic dermatitis and pemphigus were stained with safranin, and positive staining with fluorescein isothiocyanate-conjugated concanavalin A was considered to indicate the presence of glycocalyx. RESULTS: All S. aureus cells tested in skin lesions of impetigo, atopic dermatitis and pemphigus were covered with glycocalyx and formed microcolonies. The numbers of S. aureus cells in a routine light microscopy section were significantly lower than those in a frozen section that had not been dehydrated with ethanol. CONCLUSIONS: S. aureus cells generally produce glycocalyx in skin lesions of bullous impetigo, atopic dermatitis and pemphigus foliaceus, which accounts for the difficulty of removing S. aureus cells from these skin lesions. The glycocalyx may collapse during dehydration and most of the S. aureus cells may be carried away during preparation of routine light microscope sections. PMID- 12653746 TI - Widespread expression of parathyroid hormone-related peptide in melanocytic cells. AB - BACKGROUND: Parathyroid hormone-related peptide (PTH-rP) was associated with the syndrome of hypercalcaemia of malignancy. An increased serum level of PTH-rP could occur in patients with advanced melanoma. OBJECTIVES: We examined PTH-rP expression in cultured melanocytic cell lines and in lesions of melanocytic origin for associations with clinicopathological variables of disease progression. We measured the supernatant and cell lysate level of PTH-rP in cultured melanoma cells to clarify whether melanoma cells secrete PTH-rP. METHODS: PTH-rP expression was examined by reverse transcriptase-polymerase chain reaction (RT-PCR) in cultured melanocytic cell lines and by immunoperoxidase staining in 18 melanocytic naevi, 40 primary melanoma and 19 metastatic melanoma lesions. The supernatant level of PTH-rP was measured with an immunoradiometric assay. RESULTS: RT-PCR products of PTH-rP mRNA were detected in six of eight melanoma cell lines; however, neither naevus cells nor melanocytes showed positive products. On the other hand, immunohistochemical analysis showed that PTH-rP was widely expressed both in benign and malignant melanocytic lesions. In addition, PTH-rP expression was not associated with any clinicopathological variables. Cell lysate but not the supernatant of melanoma cells showed high PTH rP levels. CONCLUSIONS: These results suggest that PTH-rP was widely expressed in melanocytic cells; however, the cells did not secrete PTH-rP. PMID- 12653747 TI - Randomized comparison of photodynamic therapy with topical 5-fluorouracil in Bowen's disease. AB - BACKGROUND: Bowen's disease (BD; intraepithelial squamous cell carcinoma) is therapeutically challenging because lesions, which may be multiple, are frequently located at sites that heal poorly. There is a small risk of progression to invasive carcinoma. Photodynamic therapy (PDT) is an effective treatment for certain non melanoma skin cancers, but comparison studies with other, better-established therapies are limited. OBJECTIVES: To compare the efficacy and tolerability of PDT and topical 5-fluorouracil (5-FU) in BD. METHODS: Forty patients from two centres were randomized to either topical PDT or 5-FU. The PDT group was treated with 20% 5-aminolaevulinic acid (ALA) applied 4 h before illumination with 100 J cm-2 narrowband red light (630 +/- 15 nm). 5-FU was applied to lesions for 4 weeks. A repeat treatment cycle was performed after 6 weeks if required. Results Twenty-nine of 33 (88%) lesions treated with PDT initially responded completely, compared with 22 of 33 (67%) after 5-FU. After 12 months, two recurrences in the PDT group and six in the 5-FU group reduced complete clinical clearance rates to 82% and 48%, respectively. PDT was significantly more effective (P = 0.006, odds ratio 4.78, 95% confidence interval 1.56-14.62). In the 5-FU group, severe eczematous reactions developed around seven lesions, ulceration in three and erosions in two. No such reactions occurred following PDT. There was no difference in overall pain experienced during each therapy. CONCLUSIONS: Topical ALA-PDT is more effective than topical 5-FU in the treatment of BD, with fewer adverse events. ALA-PDT should be considered one of the first-line therapeutic options for BD. PMID- 12653748 TI - Suppression of insulin-like growth factor signalling pathway and collagen expression in keloid-derived fibroblasts by quercetin: its therapeutic potential use in the treatment and/or prevention of keloids. AB - BACKGROUND: Keloids are characterized by abnormal proliferation of fibroblasts and overproduction of collagen. Insulin-like growth factor (IGF)-I is mitogenic for fibroblasts and a stimulatory factor for collagen synthesis. OBJECTIVES: We have assessed the in vitro effects of quercetin on proliferation, collagen synthesis and the expression of the IGF system in keloid-derived fibroblasts. METHODS: Fibroblasts were isolated from earlobe keloids and exposed to quercetin at different concentrations. The inhibitory effects of quercetin on fibroblast proliferation were assayed using 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay, Western and Northern blot analyses. RESULTS: Quercetin inhibited keloid fibroblast (KF) proliferation in a dose dependent manner. Significant growth inhibition was observed on day 2 of culture. The dose required for 50% growth inhibition was approximately 25 microg mL-1. Collagen 1 expression was significantly decreased while collagen 3 was almost undetectable following quercetin treatment. Basal levels of IGF-I receptor (IGF IR) beta subunits, p85 subunit of phosphatidylinositol 3-kinase, c-Raf, phospho Raf-1, phospho-MEK 1/2, phospho-mitogen-activated protein kinase, phospho-Elk-1 and phospho-Akt-1 were significantly reduced when KF cells were exposed to quercetin for 24 h. Blocking IGF-IR activity with IGF-IR antibody or neutralizing endogenous IGF-I activity with IGF-I antibody led to significant growth inhibition suggesting the role of IGF-I in regulation of KF proliferation. CONCLUSIONS: Because the IGF system plays an important part in fibroblast cell proliferation and collagen production, the described activities of quercetin on the IGF system and collagen expression may provide a novel approach for the use of quercetin in treatment and/or prevention of hypertrophic scar and keloid. PMID- 12653749 TI - Compound heterozygosity for mutations in the hairless gene causes atrichia with papular lesions. AB - BACKGROUND: Congenital atrichias represent a complex and heterogeneous group of genodermatoses, which have been shown in several consanguineous families to result from homozygous mutations in the hairless gene (HR). OBJECTIVES: To identify the molecular basis of congenital atrichia in a non-consanguineous family. METHODS: Genetic analysis was carried out in a two-generation family with two children with congenital atrichia and one healthy child. RESULTS: We established a diagnosis of atrichia with papular lesions based on clinical and histopathological data. We identified a heterozygous 11-bp deletion (189-199del) in the two affected children and their mother. In addition, the two affected children and their father were shown to carry a non sense mutation (Q478X), which has previously been described in a Pakistani family. Haplotype analysis revealed that mutation Q478X occurred independently in the two families. CONCLUSIONS: We have identified the first case of compound heterozygosity for mutations in HR as well as the first instance of a recurrent mutation in this gene. These data further expand our understanding of the molecular pathomechanisms underlying congenital atrichias. PMID- 12653750 TI - Observer agreement in recording the clinical signs of nail disease and the accuracy of a clinical diagnosis of fungal and non-fungal nail disease. AB - BACKGROUND: Onychomycosis is increasing in incidence. To date, no studies have examined the detection of abnormalities of the nail apparatus, nor the accuracy of a clinical diagnosis of onychomycosis and non-fungal nail disease. OBJECTIVES: To assess the agreement between and within different groups of observers in detecting signs of nail disease, and to obtain information regarding clinical diagnostic skills. METHODS: An observational study was performed. Nine observers, including dermatologists, mycologists, general practitioners and a dermatology clinical assistant, completed a questionnaire containing 21 clinical signs of nail disease during examination of nine patients, five with onychomycosis and four with non-fungal nail disease. Observers were additionally requested to suggest the most likely underlying diagnosis for the nail dystrophy. Mean pair observer agreement values were calculated for each of the clinical signs, between all observers and within groups of observers. The chance-corrected agreement index, kappa, was determined. From the clinical diagnoses given, the positive predictive value of a diagnosis of fungal and non-fungal nail disease was calculated. RESULTS: There was substantial between-observer agreement on only three clinical signs: abnormal nails on both hands, abnormal toenails and abnormal fingernails. More specific signs of nail disease such as onycholysis elicited weaker agreement. All observers showed accuracy in making a clinical diagnosis of fungal nail disease, with a mean positive predictive value of 0.91, compared with 0.77 for non-fungal nail disease. CONCLUSIONS: Our results showed that agreement between observers, in recording signs of nail disease, was generally poor. The clinical diagnosis of onychomycosis was highly likely to be correct, suggesting that other criteria are being employed by individuals in reaching the diagnosis. PMID- 12653751 TI - Scleromyxoedema-like changes in four renal dialysis patients. AB - We describe four renal dialysis patients from our hospital who, over a 6-month period, developed erythematous, thickened, indurated dermal plaques. The plaques were limited to the limbs and in three patients there were associated flexion contractures. The clinical features most resembled scleromyxoedema. All patients had previously received at least one renal transplant. Histopathology of the plaques showed features of scleromyxoedema in two patients, whereas the other two showed a different picture, more suggestive of a morphoea-like process. There are important differences between our patients and classical scleromyxoedema. All four patients had normal immunoglobulins and no paraprotein was detected. Almost all cases of classical scleromyxoedema are associated with an IgGlambda paraproteinaemia. We have not yet identified an underlying cause for this cluster of cases in our hospital. It is possible that the skin changes seen may have been precipitated by an environmental agent, such as in 'toxic oil syndrome' and vinyl chloride-induced scleroderma. We discuss the differences between our patients and those with scleromyxoedema, localized or generalized morphoea and environmentally induced scleroderma. We feel that our patients show a constellation of features similar, but not identical, to scleromyxoedema. There has been only one previous report of similar patients. We believe this to be a new and distinct phenomenon. PMID- 12653752 TI - Malignant melanoma in an 8-year-old Caribbean girl: diagnostic criteria and utility of sentinel lymph node biopsy. AB - The incidence of malignant melanoma (MM) is continuing to rise, although childhood MM remains rare. We describe an 8-year-old Afro-Caribbean girl who developed a non pigmented lesion on the tip of her left thumb, which persisted despite treatment in primary care with cryotherapy. At biopsy she was found to have an acquired acral MM. She underwent amputation of the distal phalanx of her thumb, together with positive sentinel lymph node (SLN) biopsy and subsequent axillary lymph node clearance and adjuvant chemotherapy. MMs are very rare in this age and skin-type group, therefore requiring strict diagnostic criteria. These criteria include the distinction from MM mimics, especially Spitz tumours, and an appropriate use of staging techniques such as SLN biopsy to influence management. PMID- 12653753 TI - Successful treatment by double-filtration plasmapheresis of a patient with bullous pemphigoid: effects in vivo on transcripts of several genes for chemokines and cytokines in peripheral blood mononuclear cells. AB - The involvement of various cytokines and chemokines has been reported in the pathogenesis of bullous pemphigoid (BP). Double-filtration plasmapheresis (DFPP) is an effective treatment for BP but the mechanism of action remains unclear. Using semiquantitative reverse transcription-polymerase chain reaction, we examined levels of transcripts for various cytokines and chemokines in freshly isolated peripheral blood mononuclear cells in a patient with BP before and after DFPP treatment. DFPP was performed four times. Relative levels of transcripts for interleukin (IL)-8, macrophage inflammatory protein (MIP)-1alpha and IL-5, and the ratio of relative levels of transcripts for IL-4 and interferon (IFN)-gamma, were higher, before treatment, than in healthy controls, and decreased when the extent of the lesions was reduced. Relative levels of transcripts for tumour necrosis factor (TNF)-alpha and IL-4 also decreased with regression of lesions, although they were similar to or lower than the corresponding levels in healthy individuals. When eruptions recurred, relative levels of transcripts for IL-8, MIP-1alpha, RANTES (regulated upon activation normal T cell expressed and secreted), IL-2, IFN-gamma and TNF-alpha were very much higher than those prior to the recurrence, while relative levels of mRNAs for IL-4 and IL-5 did not increase. Relative levels of transcripts for IL-8, MIP-1alpha, TNF-alpha and IL-2 were lower at the end of each individual DFPP and after the four treatments than at the beginning of treatment. Our observations suggest that cytokines and chemokines produced in mononuclear cells play important roles in the pathogenesis of BP and that regulation of their expression might be involved in the therapeutic effects of DFPP in BP. PMID- 12653754 TI - Pyogenic lymphoma of the skin: a peculiar variant of primary cutaneous neutrophil rich CD30+ anaplastic large-cell lymphoma. Clinicopathological study of four cases and review of the literature. AB - Systemic anaplastic large-cell lymphoma (ALCL) in human immunodeficiency virus (HIV)-infected individuals showing an extensive infiltrate of neutrophils has been reported and referred to as 'neutrophil-rich' CD30+ ALCL. Secondary cutaneous involvement has been found in a subset of these cases. We report the clinicopathological features of four immunocompetent patients with primary cutaneous neutrophil-rich ALCL and present a new histological subtype with a dissolute growth pattern of CD30+ tumour cells. Four HIV-negative patients presented with rapidly growing solitary or multiple tumours located on the face. Ulceration of the lesions with purulent discharge was a typical finding. Various inflammatory dermatoses were considered clinically in all cases. The histological hallmark was a large number of neutrophils in the infiltrate that masked neoplastic CD30+ anaplastic cells. In two cases, a dissolute growth pattern of anaplastic tumour cells was observed. In two cases, a strong correlation between tumour growth and interleukin (IL)-8 cytokine pattern as well as the production of IL-8 by tumour cells was demonstrated. The diagnosis of neutrophil-rich ALCL is challenging clinically and histologically as the tumour cell compartment is masked by an extensive inflammatory infiltrate of neutrophils and other reactive cells such as histiocytes which may be mainly due to release of IL-8 by tumour cells. The term 'pyogenic' designates the typical feature of this distinct neutrophil-rich ALCL, namely abscess formation ('pyo-') by cytokines (IL-8) produced by tumour cells ('-genic'). The clinical behaviour of this type is the same as in primary cutaneous CD30+ ALCL with classical histological presentation. PMID- 12653755 TI - Facial lichen striatus: successful treatment with tacrolimus ointment. AB - We report a 22-year-old Japanese woman with facial lichen striatus (LS). The distribution of the lesions corresponded to that of Blaschko's lines. Histology of the lesional skin showed an inflammatory cell infiltrate around hair follicles and eccrine glands. Treatment of the linear lesions with tacrolimus ointment once or twice daily resulted in a dramatical improvement in a short time. LS is a T cell-mediated inflammatory disease and tacrolimus ointment may be an effective alternative treatment for this disease especially when the lesions are located on the face. PMID- 12653757 TI - Keratoacanthoma developing in hypertrophic lichen planus. PMID- 12653758 TI - Two new adverse effects of isotretinoin. PMID- 12653756 TI - Mycosis fungoides presenting as annular erythema. PMID- 12653759 TI - Erosive pustular dermatosis of the scalp successfully treated with calcipotriol cream. PMID- 12653760 TI - Straight hair associated with interferon alfa plus ribavirin in hepatitis C infection. PMID- 12653761 TI - Successful treatment of amyopathic dermatomyositis with topical tacrolimus. PMID- 12653762 TI - Collagenoma in Down syndrome. PMID- 12653763 TI - Trichofolliculoma: case report with immunohistochemical study of cytokeratins. PMID- 12653764 TI - Hydroxyurea-induced foot ulcer successfully treated with a topical basic fibroblast growth factor product. PMID- 12653765 TI - Raman spectroscopy analysis of protein structure of hair in patients with trichothiodystrophy. PMID- 12653766 TI - Schnitzler's syndrome: successful treatment of two patients using thalidomide. PMID- 12653767 TI - Treatment of pemphigus vulgaris with anti-CD20 monoclonal antibody (rituximab). PMID- 12653768 TI - Flutamide-induced photosensitivity: is it a forme fruste of lupus? PMID- 12653769 TI - The selective cyclooxygenase-2 inhibitor rofecoxib may improve the treatment of chronic idiopathic urticaria. PMID- 12653770 TI - Liposomal doxorubicin in the treatment of metastasized angiosarcoma. PMID- 12653771 TI - Adenolipoma of the lip. PMID- 12653772 TI - Reducing skin cancer mortality by 2010: lessons from children's sunburn. PMID- 12653773 TI - The 2-week wait: to BCC or not to BCC? PMID- 12653774 TI - Clopidogrel bisulphate-induced photosensitive lichenoid eruption: first report. PMID- 12653781 TI - The role of ultrasonographically guided puncture of the human rete testis in the therapeutic management of nonobstructive azoospermia. AB - We attempted to characterize the cells collected from the rete testis via ultrasonographically guided puncture. Unilateral puncture of the rete testis was performed in nine men with obstructive azoospermia and 51 men with nonobstructive azoospermia. All the aspirated samples from the rete testis were observed via confocal scanning laser microscope and some of them after fluorescent in situ hybridization techniques. Then therapeutic testicular biopsy was performed in the punctured testis of each man. Spermatozoa were found in all rete testis samples and all biopsy samples from obstructed men. Twenty-two nonobstructed men demonstrated absence of spermatozoa in biopsy samples. Twenty-nine nonobstructed men showed spermatozoa in biopsy material and 24 of these men (82%) had demonstrated spermatozoa in rete testis samples. There were no significant differences in fertilization and cleavage rate between intracytoplasmic sperm injection trials using biopsy spermatozoa and rete testis spermatozoa both in obstructed and nonobstructed men. Considering that puncture of the rete testis does not reduce the volume of testicular parenchyma, is less invasive and apparently causes less detrimental effect on testicular vasculature than biopsy, puncture of rete testis is recommended as first line approach for the treatment of azoospermic men. If puncture is negative for spermatozoa in nonobstructed men, biopsy is indicated. PMID- 12653782 TI - C3 in seminal plasma has no additional informative value in the diagnosis of infection/inflammation of the male genital tract. AB - The objective of this study was to determine the clinical significance of complement fraction C3 (C3c) in seminal plasma. Therefore 120 samples from randomly chosen subfertile males without signs of genital tract infection were screened for C3 and for seminal leucocytes as markers for subclinical infection/inflammation. A comprehensive semen evaluation included sperm analysis, sperm migration testing, immunocytochemical round cell differentiation to determine seminal leucocyte counts and the leucocyte ratio and semen cultures, in aliquots of the same ejaculates. C3 concentrations were significantly correlated with leucocyte counts per ml (P < 0.002) and per ejaculate (P < 0.001), and with the leucocyte ratio (P < 0.001). No association of C3 concentrations with semen quality or with the bacterial colonization of semen samples was found. The significant association with seminal leucocytes suggests that C3 might be used as an additional marker for silent male genital tract infection. In comparison with semen leucocytes, C3 screening does not reveal any further information about semen quality or infection/inflammation pathogenesis of the male genital tract. PMID- 12653783 TI - Influence of autogenous leucocytes and Escherichia coli on sperm motility parameters in vitro. AB - Urogenital infections are considered important factors in male infertility. In this in vitro study we have evaluated the impact of leucocytes in association with an artificial infection with Escherichia coli on the motility of human spermatozoa. Ejaculates and blood samples were obtained from healthy donors with normal semen parameters. Ejaculates were prepared by swim-up technique and five fractions were isolated for incubation. Leucocyte subtypes were separated from blood samples by gradient centrifugation. Purified sperm suspensions were adjusted to a concentration of 20 x 106 ml-1 and incubated with lymphocytes/ monocytes, polymorphonuclear granulocytes (PMN), and E. coli. Samples were incubated for up to 6 h at 37 degrees C. Motility analysis was performed using a computer-assisted sperm analyzer (CASA). Spermatozoa incubated with 3 x 106 PMN ml-1 revealed a significant (P=0.003) decrease in progressive motility after 2 h. This decrease remained weakly significant (P=0.024) after 4 and 6 h. Lymphocytes and monocytes had no effect on sperm motility. Spermatozoa incubated with granulocytes and E. coli demonstrated highly significant alterations in motility after 4 and 6 h of incubation (P < 0.001). The PMN indicate an effect on motility of spermatozoa under experimental conditions. However, the results suggest that bacteria are the primary agents that interfere with sperm motility. PMID- 12653784 TI - Serum levels of dimeric and monomeric inhibins and the degree of seminal alteration in infertile men with varicocele. AB - The aim of the present study was to establish the serum levels of inhibins and their relationship with the degree of seminal alteration in infertile men. Thirty six patients with varicocele (Va) and seven non-obstructive azoospermic men (Az) were included. The Va group was divided into two subgroups: Va I (sperm concentration: >20 x 106; n = 21) and Va II (sperm concentration: < 20 x 106; n = 15). Twelve fertile men were included as a control group (Co). Semen analysis and serum follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), inhibin B and Pro-alphaC levels were determined. Serum inhibin B and T levels were significantly lower and FSH and LH significantly higher in group Az when compared with the Co. Inhibin B was unable to differentiate Va I from Va II groups. However, in Va II an increase in FSH levels was observed. An inverse correlation between inhibin B and FSH, a direct correlation between inhibin B and testosterone, sperm concentration, motility and morphology were found. No such correlations were seen when only the Va group was analysed. The lack of correlation between serum levels of inhibin B, gonadotrophins, sperm concentration and seminal parameters observed in Va, adds other factor to the complex pathophysiology of varicocele. Finally, further studies are needed to elucidate if oligozoospermic patients with varicocele have also an impaired negative feed-back mechanism that regulates FSH synthesis and secretion. PMID- 12653785 TI - Induction of the acrosome reaction in bull spermatozoa with plasmin. AB - Proteolytic enzymes appear to have an essential role in multiple phases of mammalian fertilization. Plasmin, the active enzyme of the plasminogen activation system that stimulates fibrinolysis and proteolysis has a less well-documented role in reproduction. The current study was conducted to investigate the effect of the active protease, plasmin, on the ability of bovine sperm to undergo the acrosome reaction. Aliquots of freshly ejaculated bull sperm were incubated in capacitating conditions with 10 microg ml-1 of heparin for 4 h. Every 2 h an aliquot of spermatozoa was exposed to lysophosphatidylcholine (100 microg ml-1) or 0, 0.1, 1, 10 and 100 mU of plasmin to induce the acrosome reaction in capacitated spermatozoa. Plasmin increased the percentage of live acrosome reacted sperm after 4 h of incubation in the capacitation medium. Viability was not affected by any of the treatments. This study provides new information on bovine acrosome reaction during in vitro incubation with plasmin and indicates that this protease may participate in the proteolytic events that accompany fertilization. PMID- 12653786 TI - The effect of colloid osmotic pressure in human spermatozoa exposed to hypoosmotic conditions. AB - The use of a protein source such as serum and albumin had been extensively employed as supplements of culture media for handling and culture of gametes and embryos. Protein molecules behave as colloids in solution and contribute to the osmotic pressure of fluids. The interaction of proteins in solution and spermatozoa needs to be assessed in order to determine their possible role in osmoregulation. The aim of this study was to assess possible osmoregulatory mechanisms of protein supplementation against exposure to hypoosmotic conditions by assessing the sperm's response to those environments. A stock hypoosmotic solution (HOS) was prepared by using a mixture of fructose and sodium citrate and adjusted to an osmotic pressure of 150 mOsm l-1. Another stock solution was prepared by diluting a preparation of synthetic serum supplement [SSS; 6% (v/v) total protein] with distilled water to obtain an osmotic pressure of 150 mOsm l-1 (hypoosmotic SSS or H-SSS). Three additional solutions were prepared by mixing the stock HOS and H-SSS solutions in the following proportions (v/v): (i) 75% H SSS/25% HOS, (ii) 50% H-SSS/50% HOS and (iii) 25% H-SSS/75% HOS. Aliquots of washed spermatozoa from 18 men were diluted 1 : 10 (v/v) with each of the testing solutions and incubated for 60 min. Specimens were assessed on wet mounts for total and specific swelling patterns. Swelling patterns were classified as maximal (>50% tail length swollen) and minimal (<50% tail length swollen) swelling with or without associated sperm motility. The major finding of this study was that increasing the concentration of protein supplementation resulted in a decrease in the proportion of maximal sperm tail swelling patterns and an increase in the proportion of minimal tail swelling patterns. A proportion of spermatozoa which exhibited minimal swelling patterns were still motile in all solutions tested, and the percentage of those spermatozoa increased as the protein supplementation was also increased in the testing solutions. Incorporation of protein supplementation as described in this study delays the effect of sperm swelling in hypoosmotic conditions. PMID- 12653787 TI - Comparison of fine-needle aspiration and open biopsy of testis in sperm retrieval and histopathologic diagnosis. AB - This study aimed to compare the diagnostic value of testicular fine-needle aspiration (FNA) with open biopsy in infertile males with azoospermia or severe oligozoospermia and to evaluate the reliability of testicular sperm extraction by FNA. A total of 76 testes of 40 patients, 34 with azoospermia, four oligozoospermia and two patients who underwent orchidectomy as a result of cancer of prostate were included. Detailed clinical and laboratory examinations were performed and two semen analyses were obtained from each patient. A 20-ml 26 gauge 13-mm needle was used for FNA and smears were stained with May-Grunwald Giemsa and Papanicolaou stain. An open biopsy was performed in each patient after FNA and the samples were stained with haematoxylen-eosin. Smears and histological stains were examined and compared under light microscopy by the same pathologist. In 69 of the 76 testes (90%) FNA cytology results agreed with the histology. In four testes, the aspirate was unsatisfactory and in three testes, spermatocytic arrest was found cytologically while subsequent biopsies revealed diffuse fibrosis. In 15 of 16 patients (93.7%) with normal and hypospermatogenesis, spermatozoa had been extracted by FNA. Spermatozoa could not be obtained after neither FNA nor open biopsy in the remaining 24 patients. Testicular FNA in infertile males is a simple, reliable and minimally invasive diagnostic tool. It is as effective as open biopsy for testicular sperm extraction and good results can be achieved in experienced hands. PMID- 12653790 TI - Aging and asthma: pathophysiological mechanisms. PMID- 12653788 TI - Levels of interleukin-6 and interleukin-8 in seminal fluid of men attending an andrological clinic. AB - The presence of interleukins (IL) and other cytokines in seminal plasma was demonstrated in the literature. In particular, the levels of IL-6 were found to be related to male accessory gland inflammation. The close correlation to leucocyte count indicates a production of interleukins from the leucocytes and by the prostate gland. No relation of IL-6 levels to spermatogenic activity was quoted in the literature. We measured IL-6 and IL-8 in 454 men and compared the values with seminal parameters. The mean values of IL-6 30.7 +/- 101.2 pg ml-1 and IL-8 2023 +/- 1721 pg ml-1. The correlation analysis revealed a significant correlation of IL-6 and/or IL-8 to age, total fructose, immunoglobulin G (IgG) concentration and leucocyte count. The significant correlation of IL-6 and fructose levels indicates that also the seminal vesicles take part in the production of seminal IL-6. No correlation of the two interleukins measured to sperm parameters occurred. The calculation of a single harmonic trend revealed a significant trend over the year of the levels of IL-6 with a maximum in December and a peak-to-trough variation of 33% of the mean. It may be the consequence of a higher frequency of seminal tract inflammations in autumn and winter. PMID- 12653791 TI - An update on the diagnosis and treatment of sinusitis and nasal polyposis. PMID- 12653792 TI - Requirements for medications commonly used in the treatment of allergic rhinitis. European Academy of Allergy and Clinical Immunology (EAACI), Allergic Rhinitis and its Impact on Asthma (ARIA). PMID- 12653793 TI - Nuclear transport factor 2 represents a novel cross-reactive fungal allergen. AB - BACKGROUND: Ubiquitously occuring moulds are important allergenic sources known to elicit IgE-mediated allergic diseases and to share cross-reactive allergens. Limited information is available about the molecular structures involved in cross reactivity. We aimed to clone and characterize cross-reactive mould allergens. METHODS: Phage surface-displayed Alternaria alternata and Cladosporium herbarum cDNA libraries were screened using sera from Aspergillus fumigatus-sensitized patients. Inserts encoding putative allergens were sequenced, and recombinant proteins used to demonstrate cross-reactivity by inhibition experiments and skin test. Three-dimensional homology models of cloned putative nuclear transport factor 2 (NTF2) were constructed based on known NTF2 structure to corroborate the functional and structural properties of the novel allergens. RESULTS: After six rounds of affinity selection, the libraries were enriched for clones displaying allergens. Sequencing of inserts showed that some clones derived from Alternaria alternata and Cladosporium herbarum contain open reading frames predicting proteins of 124 and 125 amino acids corresponding to NTF2. The recombinant proteins were able to bind and cross-inhibit IgE binding and to elicit type I skin reactions in mould-sensitized individuals, demonstrating the allergenicity of the proteins. CONCLUSIONS: NTF2 represents a novel cross-reactive fungal allergen as demonstrated by sequence homology, three-dimensional modelling, inhibition experiments and skin test reactivity. PMID- 12653794 TI - Effect of ciclesonide on allergen challenge in subjects with bronchial asthma. AB - BACKGROUND: The aim of this clinical trial was to investigate whether repeated inhalation of the new inhaled steroid ciclesonide reduces the early-phase (EAR) and late-phase (LAR) reactions after allergen challenge in patients with mild allergic asthma. Also, this study provides further data on safety and tolerance of ciclesonide. METHODS: The study was designed as a double-blind placebo controlled randomized crossover trial. Following a baseline period, patients were randomized to either of two treatment sequences (ciclesonide/placebo, placebo/ciclesonide) each of which lasted for one week and were separated by 3-5 weeks from the alternate treatment sequence. Patients received 800 micro g ciclesonide twice daily by means of a Cyclohaler. At the end of each treatment patients were subjected to an allergen challenge. RESULTS: Thirteen asthmatic patients (mean FEV1 of 91% predicted) who experienced an EAR and LAR after allergen challenge participated in the study. The time-average FEV1 decreases 0-2 h (2-12 h) after allergen challenge as measure of the EAR (LAR) were significantly reduced (P < 0.05, one-sided) from 0.426 L to 0.233 L (EAR) and from 0.443 L to 0.213 L (LAR), respectively. Thus, the study results suggest that ciclesonide significantly lowered the extent of EAR and LAR compared to placebo. Ciclesonide was well tolerated and no drug-related adverse events were reported. Cortisol excretion in 24-h urine showed no significant difference between ciclesonide and placebo. CONCLUSIONS: The study supports the efficacy and safety of ciclesonide. PMID- 12653795 TI - Bronchial hyperresponsiveness and airway inflammation in adolescents with asymptomatic childhood asthma. AB - BACKGROUND: About 70% of childhood asthmatics become free of asthma-related symptoms during adolescence. Little is known about bronchial hyperresponsiveness (BHR) and airway inflammation in young adults with "outgrown" childhood asthma. METHODS: We studied 61 nonsmoking medical students (18 intermittent mild asthmatics, 23 students with outgrown childhood asthma but free of asthma-related symptoms for 10 years (asymptomatic asthmatics) and 20 healthy students). BHR and lung function were measured, and induced sputum samples analyzed for eosinophil count, eosinophilic cationic protein (ECP), granulocyte-macrophage colony stimulating factor (GM-CSF), and tumor necrosis factor-alpha (TNF-alpha). RESULTS: BHR was still present in most asymptomatic asthmatics, but it was milder compared with healthy students. Only three subjects with previous asthma had no BHR and no signs of airway inflammation. Percentages of eosinophil, and ECP, TNF alpha and GM-CSF concentrations in induced sputum of mild asthmatics and asymptomatic asthma groups were higher than in the healthy group. In asymptomatic asthmatics group, the duration of asthma, sputum eosinophil percentage, and the level of TNF-alpha in sputum correlated significantly with BHR. CONCLUSIONS: Only a few subjects with longstanding asymptomatic asthma could be considered as cured; most asymptomatic asthmatics continued to exhibit BHR and signs of airway inflammation. The outcome of childhood asthma and BHR was associated with the degree of airway inflammation and the duration of childhood asthma. PMID- 12653796 TI - The control of asthma in Italy. A multicentre descriptive study on young adults with doctor diagnosed current asthma. AB - BACKGROUND: Few data are available on the management of asthma in the general population. The aim of this study was to evaluate the level of asthma control in Italian patients, a decade after the publication of the international guidelines. MATERIALS AND METHODS: Within the framework of a multicentre, population-based study on people aged 20-44 years, 18 873 subjects replied to a postal screening questionnaire (response rate = 72.7%) on the presence of asthma symptoms and exacerbations, and their impact on daily life. All subjects reporting having ever had a doctor diagnosis of asthma and either taking medicine for asthma when interviewed or having had an asthma attack in the last 12 months, were considered current asthmatics. RESULTS: Of the screened subjects, 649 (3.4%) were current asthmatics. Of these, only 14% did not report exacerbations or had been symptom free in the last 12 months. More than 20% of current asthmatics had their daily life activities seriously impaired and 54 patients (8%) had at least one hospital/emergency department admission as a result of asthma exacerbation in the last 12 months. The life impairment and the rates of hospitalization significantly increased as the control of the symptoms worsened. The use of asthma drugs was quite common in current asthmatics: 586 (90.2%) reported having been under pharmacological treatment in the last 12 months. Only 63 patients (10%) had the disease under control (neither symptoms nor life impairment): they had a significantly higher percentage of drug use (100%vs 89%) and of daily use prescriptions (50%vs 36%) than non/moderately controlled asthmatics. Poorly controlled asthmatics had a significantly higher percentage of women (63%vs 44%), of patients with the coexistence of chronic cough and phlegm (47%vs 30%) than moderately/well-controlled asthmatics. The comparison between our data and similar data collected in 1991 in Italy showed that the use of asthma drugs increased by about 12%, while the control of symptoms did not improve in the last decade. CONCLUSION: Despite the high percentage of drug users, the control of symptoms and exacerbations was overall poor in Italy and resulted in a heavy individual and social burden, pointing out that the guideline goals have far from been reached in Italy. PMID- 12653797 TI - Maternal oral contraceptive use and atopic diseases in the offspring. AB - BACKGROUND: This study examined the association of maternal oral contraceptive (OC) use - before and after birth - and atopic manifestations in the offspring. METHODS: A total of 2754 East German children aged 5-14 years participated in a cross-sectional survey in 1998-99. The standardized parental questionnaire in 1998-99 included data on atopic diseases, socio-economic factors, parental atopy and maternal OC use. Specific immunoglobulin E against common inhalant allergens was measured by radioallergosorbent test (RAST). RESULTS: Maternal OC use before birth was associated with a higher risk of atopic diseases in the offspring compared with children of mothers who had never taken OC [asthma: odds ratio (OR) 1.6; 95% confidence interval (CI): 0.9-3.0; allergic rhinitis: OR 1.5; CI: 0.96 2.2; atopic eczema: OR 2.6; CI: 1.6-4.3; atopic sensitization: OR 1.5; CI: 0.97 2.2]. However, the effect estimates for maternal OC use after birth compared with the never users showed quite similar effects for these atopic conditions. No relations were observed between the prevalences of atopic diseases and maternal age at beginning of OC use, the duration of OC use, the type of contraceptive or maternal age at birth. CONCLUSION: This study raises doubts in a true biological association between OC use and atopic diseases. PMID- 12653798 TI - National survey on the management of rhinopathies in asthma patients by French pulmonologists in everyday practice. AB - BACKGROUND: Epidemiological and pathophysiologic studies have shown that allergic rhinitis and asthma often occur in combination. The internationally developed ARIA position paper (Allergic Rhinitis and its Impact on Asthma) recently offered recommendations on allergic rhinitis. As part of this new report and prior to its diffusion, we investigated the management of rhinopathies in asthma patients by pulmonologists in their everyday practice. METHODS: From March to June 2000, 477 (48%) French pulmonologists in office-based practice participated in the survey. They were asked to include their first five asthmatic adult patients. In addition to descriptive statistics, univariate and multivariate analyzes were performed. RESULTS: We studied 1623 patients with varying severity of asthma (sex ratio 0.9; median age 35 years). The pulmonologists reported rhinopathy in 76.6% of these, with a chronic course in 91%. Among the patients, 67.1% reported rhinopathy. The diagnosis was allergic rhinitis in 66.2% of participants and nasal polyposis in 10.1%. Examination of the nasal cavities was performed by the pulmonologists themselves in 56.2% of patients. Imaging of the sinuses was performed radiographically in 55.3% of enrolled patients and/or by computed tomography in 17.2%. Referral to an ENT specialist occurred for 21.6% of patients, being more common for patients with rhinitis that failed to respond to medical therapy (although some pulmonologists referred their patients routinely). CONCLUSIONS: The high prevalence of rhinopathies in asthma patients requires that these conditions are recognized and managed by pulmonologists. Thus, our findings support one of the central messages contained in the new ARIA guidelines-asthma patients should be investigated routinely for rhinitis and other rhinopathies. PMID- 12653799 TI - Altered intracellular expression of the chemokines MIP-1alpha, MIP-1beta and IL-8 by peripheral blood CD4+ and CD8+ T cells in mild allergic asthma. AB - BACKGROUND: The ability of chemokines to regulate Th1 and Th2 responses suggests a role in the pathogenesis of atopic disorders such as allergic asthma where Th2 response dominance has been observed. Although the impact of allergic asthma on local chemokine production in the lung has been the subject of investigation, little is know about the influence of disease progression on peripheral chemokine production. We now report use of whole blood culture and flow cytometry to assess the influence of mild allergic asthma on peripheral T-cell chemokine expression. METHODS: Study participants included patients with mild allergic asthma (n = 7) and nonasthmatic controls (n = 7). Following in vitro stimulation of peripheral venous blood with phorbol 12-myristate acetate (PMA) and ionomycin, flow cytometry was used to estimate the percentage of CD4+ and CD8+ T cells producing a number of chemokines, including macrophage inflammatory proteins MIP-1alpha and MIP-1beta, RANTES (regulated on activation, T-cell expressed and secreted), monocytic chemotactic protein-1 (MCP)-1, and interleukin (IL)-8, or the cytokines interferon (IFN)-gamma and IL-4. Serum levels of MIP-1alpha, MIP-1beta, RANTES, MCP-1, IL-8, IFN-gamma and IL-4 were also assessed by quantitative ELISA. RESULTS: Intracellular expression of MIP-1beta by CD4+ and CD8+ T cells from allergic asthmatics was significantly reduced in comparison to that observed for nonasthmatics (median = 2.29% (1.75-3.50) vs 4.57% (3.38-6.64), P = 0.05; 14.20% (13.18-17.88) vs 44.10% (30.38-48.70), P = 0.01). Similarly, intracellular expression of MIP-1alpha by CD8+ T cells from allergic asthmatics was also significantly lower (3.67% (1.17-5.42) vs 17.10% (4.97-20.43), P = 0.05). Conversely, IL-8 expression by both CD4+ and CD8+ T cells from allergic asthmatics demonstrated significant enhancement (9.93% (7.77-11.28) vs 4.14% (3.61-7.11), P = 0.05; 8.40% (6.97-10.04) vs 4.98% (3.37-6.08), P = 0.05). Examination of intracellular IFN-gamma and IL-4 revealed no significant difference in the expression of either cytokine by CD4+ T-cells from allergic asthmatics and nonasthmatics. In contrast, expression of IFN-gamma was significantly reduced in CD8+ T-cells from allergic asthmatics (24.60% (21.08 32.50) vs 48.40% (41.50-55.28), P = 0.01). CONCLUSIONS: The occurrence in mild allergic asthma of peripheral T-cell chemokine expression suggestive of a diminished Th1 response, coinciding with marginal change in cytokine profiles indicative of a Th2 response bias, confirms the importance of chemokine involvement in the etiology of allergic asthma. The ability to use whole blood culture to estimate chemokine expression in T cell subsets may ultimately provide a practical means to evaluate disease status and to monitor early intervention therapies which target chemokines. PMID- 12653800 TI - Molecular cloning, purification, and IgE-binding of a recombinant class I chitinase from Hevea brasiliensis leaves (rHev b 11.0102). AB - BACKGROUND: Class I chitinase in natural rubber latex (NRL) has been assumed to be an important allergen, especially concerning its cross-reactivity with fruits like avocado and banana. OBJECTIVES: The present study aimed to produce a recombinant latex class I chitinase from Hevea brasiliensis leaves and to study its immunoglobulin (Ig)E-binding reactivity. METHODS: A class I chitinase specific complementary DNA from H. brasiliensis leaves was synthesized, subcloned, sequenced and overexpressed in fusion with the maltose-binding protein (MBP) in Escherichia coli. The IgE-binding reactivity of this protein was studied by the Pharmacia CAP System and by immunoblot experiments using sera from latex allergic patients. RESULTS: The rHev b 11.0102 was found to have a length of 295 amino acid residues and contains an N-terminal hevein-like domain with a 56% homology to hevein. Analysis by the CAP method revealed the presence of rHev b 11.0102-specific IgE antibodies in 17 of 58 sera (29%) of IgE-mediated latex allergic subjects tested. Immunoblot analysis of the MBP-rHev b 11.0102 fusion protein and the MBP carrier protein as a negative control confirmed the IgE reactivity of rHev b 11.0102. CONCLUSION: Due to its IgE-reactivity rHev b 11.0102 represents an allergen of intermediate prevalence in NRL. Its property to cross-react with certain fruits makes it an important supplement in the diagnostic panel of recombinant NRL allergens. PMID- 12653801 TI - Staphylococcal enterotoxin induced IL-5 stimulation as a cofactor in the pathogenesis of atopic disease: the hygiene hypothesis in reverse? AB - BACKGROUND: The incidence of Staphylococcus aureus (S. aureus) colonization on the skin of patients with atopic eczema/dermatitis syndrome (AEDS) is approximately 90% and a variety of evidence implicates epidermal staphylococcal infection as a pathogenic factor in atopic dermatitis. However, the mechanism(s) underlying the effects of this organism in the disease process are unclear. The cellular responses of AEDS suffers and asymptomatic atopic individuals to bacterial superantigens (SAg) were investigated in an attempt to elucidate the role of staphylococcal enterotoxin B (SEB) in atopic disease. METHODS: Peripheral blood mononuclear cells (PBMC) were isolated from normal nonatopic adults, asymptomatic atopic individuals, patients with active AEDS and patients with active allergic asthma. The cells were cultured for 24 or 96 h with house dust mite (HDM), SEB and phytohaemagluttinin (PHA), and the supernatants were assayed for cytokine levels. RESULTS: Staphylococcal enterotoxin B selectively stimulates the production of interleukin (IL)-5 in AEDS sufferers but not in asymptomatic atopics or nonatopics. Additionally, we observed comparable susceptibility to the IL-5-stimulatory effects of SEB in allergic asthmatics. CONCLUSIONS: Given the central role of IL-5-driven eosinophilia in progression from mild atopy to severe disease, these findings provide a plausible mechanism for the AEDS-promoting effects of staphylococcal SAg. Staphylococcal enterotoxin B may also have a similar role in atopic respiratory disease. PMID- 12653802 TI - Comparison of standard and modified SPT method. AB - BACKGROUND: The aim of our study was to compare clinical significance and economical factors of commonly used standard skin prick tests (SPT) method with new, modified and more handy way of performing SPT. METHODS: Twenty-one subjects [with an average age of 23 years (SD +/- 2.72)] with known sensitivity to examined allergen extracts were tested with histamine, negative control solution and standard allergen extracts (Allergopharma, Reinbeck, Germany). SPT were applied to the volar surface of randomly assigned forearm with two methods: standard technique using lancet needle to prick the epidermis under small drop of extract and tested method using lancet needle which was dipped in extract before prick. In this method all extracts were placed in marked, small, plastic chambers. RESULTS: Mean wheal to histamine was 5.49 (standard method) and 5.01 (modified method). There was no statistical difference between the standard and modified method for size of wheal to histamine (P = 0.654) and to allergen extracts (n = 36, P = 0.824, Wilcoxon test). Economical analysis revealed that examined SPT technique is cheaper than standard method. CONCLUSION: Examined method may be considered to be an alternative for performing SPT as it is safe, cheaper and more convenient than standard technique. SUMMARY STATEMENT: Comparison of standard and modified SPT method revealed the new method to be more economical. PMID- 12653803 TI - Oral allergic syndrome after ingestion of custard apple. PMID- 12653804 TI - IP-10 in atopic dermatitis. PMID- 12653805 TI - Late-type allergy to Ultravist (iopromid). PMID- 12653806 TI - Contact allergy to phenylpentanedione. PMID- 12653807 TI - Methadone syrup injection in Australia: a sentinel finding? PMID- 12653808 TI - Methadone-related and heroin-related deaths among opiate users: methadone helps save lives. PMID- 12653809 TI - Conversation with Ambros Uchtenhagen. PMID- 12653810 TI - Longer-term primary prevention for alcohol misuse in young people: a systematic review. AB - OBJECTIVE: To identify and summarize rigorous evaluations of psychosocial and educational interventions aimed at the primary prevention of alcohol misuse by young people aged up to 25 years, especially over the longer term (>3 years). METHODS: Cochrane Collaboration Systematic Review. DATA SOURCES: A comprehensive search of 22 databases and recursive checking of bibliographies for randomized and non-randomized controlled trials and interrupted time-series studies. MAIN OUTCOME MEASURES: Objective or self-report measures of alcohol use and misuse. RESULTS: Fifty-six studies were selected for inclusion in the systematic review. Twenty of the 56 studies showed evidence of ineffectiveness. No firm conclusions about the effectiveness of prevention interventions in the short- and medium term were possible. Over the longer term (>3 years), the Strengthening Families Programme (SFP) showed promise as an effective prevention intervention. The Number Needed to Treat (NNT) for the SFP over 4 years for three alcohol initiation behaviours (alcohol use, alcohol use without permission and first drunkenness) was 9 (for all three behaviours). One study also highlighted the potential value of culturally focused skills training over the longer-term (NNT = 17 over 3.5 years for 4+ drinks in the last week) CONCLUSIONS: (1) Research into important outcome variables needs to be undertaken; (2) the methodology of evaluations needs to be improved; (3) the SFP needs to be evaluated on a larger scale and in different settings; (4) culturally focused interventions require further development and rigorous evaluation; and (5) an international register of alcohol and drug misuse prevention interventions should be established and criteria agreed for rating prevention interventions in terms of safety, efficacy and effectiveness. PMID- 12653811 TI - Prevalence and correlates of intravenous methadone syrup administration in Adelaide, Australia. AB - AIMS: The aims of this study were to determine the prevalence of methadone syrup injecting in Adelaide, South Australia and to characterize methadone injectors, including their heroin use and risk behaviours associated with heroin overdose. DESIGN: Cross-sectional design. SETTING: Community setting, principally metropolitan Adelaide. PARTICIPANTS: Current heroin users (used heroin in the last 6 months), recruited through snowballing. MEASUREMENTS: Structured questionnaire. FINDINGS: Of 365 participants, 18.4% reported having ever injected methadone syrup and 11.0% had injected methadone in the last 6 months. Those that had injected methadone were more likely to be male, and were more likely to be receiving methadone maintenance. They were also maintained on higher doses of methadone than subjects not injecting methadone. A history of methadone injection was associated with more heroin overdose experiences and greater dependence on heroin. Methadone injectors were also more likely to engage in risky behaviours associated with heroin overdose, including using heroin when no other people were present, not trial-tasting new batches of heroin and polydrug use. CONCLUSIONS: Methadone syrup injectors appear to be at greater risk of a series of harms than subjects not injecting methadone. The prevalence of methadone syrup injecting in Adelaide, South Australia was 11%, which was lower than prevalence in Sydney, New South Wales, but higher than in Melbourne, Victoria. Jurisdictional differences concerning the prevalence of methadone syrup injecting may reflect differing policies by each state to methadone dispensing. PMID- 12653812 TI - Trends in drug overdose deaths in England and Wales 1993-98: methadone does not kill more people than heroin. AB - AIMS: To test the hypothesis that methadone is responsible for a greater increase in overdose deaths than heroin, and causes proportionally more overdose deaths than heroin at weekends. DESIGN AND SETTING: Multivariate analysis of 3961 death certificates mentioning heroin, morphine and/or methadone held on the Office for National Statistics drug-related poisoning mortality database from 1993 to 1998 in England and Wales. MEASUREMENTS: Percentage increase in deaths by year by drug, odds ratio (OR) of dying at the weekend from methadone-related overdose compared to dying from heroin/morphine overdose. FINDINGS: From 1993 to 1998, annual opiate overdose deaths increased from 378 to 909. There was a 24.7% (95% confidence interval (CI) 22-28%) yearly increase in heroin deaths compared to 9.4% (95% CI 6-13%) for methadone only. This difference was significant (P < 0.001 by test of interaction) after adjustment for sex, age group, polydrug use, area of residence and underlying cause of death. The largest number of deaths occurred on Saturday (673). The OR of death from methadone overdose on Saturday and Sunday was 1.48 (95% CI 1.29-1.71) for methadone-only deaths compared to dying from heroin/morphine at the weekend after adjustment for other covariates, but the OR was not significant (1.09, 95% CI 0.95-1.25) if the weekend was defined as Friday and Saturday. CONCLUSIONS: There was no evidence that the threefold increase in deaths over time was due to methadone. There was equivocal support only for the hypothesis that there was an excess of deaths from methadone at weekends. Increased interventions to prevent overdose among injectors in England and Wales are long overdue. PMID- 12653813 TI - Evaluation of the effects of lofexidine and clonidine on naloxone-precipitated withdrawal in opioid-dependent humans. AB - AIMS: To examine the efficacy of lofexidine, an alpha2 adrenergic agonist, to suppress opioid withdrawal symptoms in opioid-dependent humans under in-patient laboratory conditions by using a naloxone challenge procedure. DESIGN: Randomized, within-subject, cross-over design with drug administration taking place under double-blind and triple-dummy conditions. SETTING: A 14-bed in patient hospital research unit dedicated to the conduct of behavioral pharmacology studies. PARTICIPANTS: Eight healthy adult volunteers (two female/six male) with histories of polysubstance abuse and current physical dependence on opioids. INTERVENTION: Participants were stabilized onto methadone and maintained on 30 mg/day, p.o. throughout the study. Oral placebo, lofexidine (0.4, 0.8 and 1.6 mg, p.o.) and clonidine (0.1 and 0.2 mg, p.o.) were each tested as pre-treatments once in combination with each of three intramuscular naloxone doses (0, 0.1 and 0.3 mg, i.m.) during 18 separate experimental sessions. MEASUREMENTS: An array of physiological indices (e.g. heart rate, blood pressure, pupil diameter) as well as a number of subjective and observer-rating scales sensitive to opioid withdrawal effects. FINDINGS: As expected, lofexidine and clonidine both produced dose-related decreases in blood pressure and heart rate but few subjective effects; naloxone increased opioid withdrawal signs and symptoms in a dose- and time-dependent fashion. Although both lofexidine and clonidine reduced the cardiovascular response to naloxone challenge, close inspection of the data reveal that this occurred only to the extent that baseline physiological parameters were reduced, while neither drug significantly modified the overall magnitude of the response to naloxone. Moreover, neither lofexidine nor clonidine suppressed the subjective discomfort of opioid withdrawal or significantly reduced other autonomic signs of opioid withdrawal, such as lacrimation or rhinorrhea. CONCLUSIONS: These data suggest that lofexidine is well tolerated even at supratherapeutic acute doses. However, its failure to modify most signs and symptoms of opioid withdrawal suggest that its effective use in spontaneous withdrawal will require concomitant medications for improved therapeutic response. PMID- 12653814 TI - Buprenorphine versus methadone maintenance therapy: a randomized double-blind trial with 405 opioid-dependent patients. AB - AIMS: To assess the efficacy of buprenorphine compared with methadone maintenance therapy for opioid dependence in a large sample using a flexible dosing regime and the marketed buprenorphine tablet. DESIGN: Patients were randomized to receive buprenorphine or methadone over a 13-week treatment period in a double blind, double-dummy trial. SETTING: Three methadone clinics in Australia. PARTICIPANTS: Four hundred and five opioid-dependent patients seeking treatment. INTERVENTION: Patients received buprenorphine or methadone as indicated clinically using a flexible dosage regime. During weeks 1-6, patients were dosed daily. From weeks 7-13, buprenorphine patients received double their week 6 dose on alternate days. MEASUREMENTS: Retention in treatment, and illicit opioid use as determined by urinalysis. Self-reported drug use, psychological functioning, HIV-risk behaviour, general health and subjective ratings were secondary outcomes. FINDINGS: Intention-to-treat analyses revealed no significant difference in completion rates at 13 weeks. Methadone was superior to buprenorphine in time to termination over the 13-week period (Wald chi 2 = 4.371, df = 1, P = 0.037), but not separately for the single-day or alternate-day dosing phases. There were no significant between-group differences in morphine-positive urines, or in self-reported heroin or other illicit drug use. The majority (85%) of the buprenorphine patients transferred to alternate-day dosing were maintained in alternate-day dosing. CONCLUSIONS: Buprenorphine did not differ from methadone in its ability to suppress heroin use, but retained approximately 10% fewer patients. This poorer retention was due possibly to too-slow induction onto buprenorphine. For the majority of patients, buprenorphine can be administered on alternate days. PMID- 12653815 TI - Short-term outcomes after brief ambulatory opioid detoxification with buprenorphine in young heroin users. AB - AIMS: This study examines the outcomes at 1, 3 and 6 months after a very brief outpatient detoxification with buprenorphine in 18-25-year-old heroin users. DESIGN: Prospective follow-up study. SETTING: Outpatient drug treatment clinic, providing brief detoxification in downtown Baltimore, Maryland, USA. PARTICIPANTS: One hundred and twenty-three subjects between 18 and 25 years old; 56% male; 95% Caucasian; seeking detoxification; living in Baltimore City and five surrounding counties. INTERVENTION: Detoxification with buprenorphine over 3 days. Follow-up at 1, 3 and 6 months. MEASUREMENTS: Drug use history, the Addiction Severity Index at baseline and follow-up, urine drug screens, evaluation of the detoxification experience. FINDINGS: By self-report, 37% of the total sample were not currently using heroin at 1 month, 32% at 3 months and 29% at 6 months, and 6.7%, 10.1% and 11.8% had an opioid negative urine test at 1, 3 and 6 months, respectively. There was a significant reduction from the baseline in mean Addiction Severity Index drug use composite score, as well as the mean number of days of heroin and cocaine use during past 30 days, that was sustained over the three follow-up points. Engagement in aftercare was generally poor. CONCLUSIONS: The findings show a reduced frequency and intensity of drug use, suggesting a possible role for brief outpatient detoxification in reducing the severity of dependence for some younger heroin users who may not yet be ready to engage in long-term abstinence-oriented or opioid substitution treatments. PMID- 12653816 TI - Use of morphine and 6-monoacetylmorphine in blood for the evaluation of possible risk factors for sudden death in 192 heroin users. AB - AIMS: To detect risk factors for sudden death from heroin injection. DESIGN: Evaluation of data from forensic investigations of all fatal cases of suspected heroin death in a metropolitan area. Only cases with detectable morphine and 6 monoacetylmorphine (6-MAM) in blood were included in order to select heroin intoxication cases. SETTING: Stockholm, Sweden. MEASUREMENTS: Autopsy investigation and toxicological analysis of blood and urine; and police reports. FINDINGS: In two-thirds of the 192 cases, death occurred in public places, and mostly without any time delay. Blood concentrations of morphine ranged from 50 to 1200 ng/g, and of 6-MAM from 1 to 80 ng/g. Codeine was detected in 96% of the subjects. In the majority of cases the forensic investigation indicated polydrug use, the most common additional findings being alcohol and benzodiazepines. However, in one-quarter of the cases other drug combinations were found. Previous abstinence from heroin and use of alcohol were identified as risk factors. For 6 MAM there was also a correlation with the presence of THC and benzodiazepines. Despite a high frequency of heart abnormalities (e.g. myocarditis and focal myocardial fibrosis), these conditions did not correlate with morphine or 6-MAM blood concentrations. CONCLUSIONS: We confirm that alcohol intake and loss of tolerance are risk factors for death from heroin use, whereas no connection to heart pathology was observed. Further, prospective, studies should focus on other possible risk factors. PMID- 12653817 TI - Estimating prevalence of problem drug use at national level in countries of the European Union and Norway. AB - OBJECTIVES: This paper will present the most recent estimates for problematic drug use in European Member States and explore the problems of comparability. METHODS: Estimates of problematic drug use, derived according to agreed guidelines, were collected from all EU countries and Norway. Methods included multipliers based on treatment, police, mortality or AIDS/HIV data, the capture recapture method and the multivariate indicator method. Prevalence estimates were transformed into rates per 1000 population aged 15-64 years. RESULTS: Target populations varied according to data selection. Estimates for six partially overlapping types of drug use could be identified: 'problem opiate use', 'problem opiate or cocaine use', 'problem amphetamine or opiate use', 'problem drug use', '(current) injecting' and 'life-time injecting'. Rates of injectors ranged from 2.6 in Germany to 4.8 in Luxembourg; rates in Austria, Denmark, Finland, Portugal and the United Kingdom fell within this range. For problem opiate use, a group of high prevalence countries were found with average rates exceeding six cases (Italy, Luxembourg, Spain and the United Kingdom) and a group with lower prevalence with average rates close to three cases (Austria, Germany, Ireland, the Netherlands). Rates obtained for France (problem opiate or cocaine use), Finland and Sweden (problem amphetamine or opiate use) are not directly comparable and fall between these rates. CONCLUSIONS: Cross-national comparisons should be made with care and estimated target populations may vary greatly between countries. For estimating various forms of problem drug use at national level, a multi-method approach is recommended. PMID- 12653818 TI - Delphi survey of experts' opinions on strategies used by community pharmacists to reduce over-the-counter drug misuse. AB - AIM: To explore the views of experts within the fields of pharmacy and addiction on the value of current strategies and possible alternatives and to reach an agreement on best practice in the sale of over-the-counter (OTC) medicines which are liable to misuse. DESIGN: Using a modified Delphi approach, an anonymous, international, three-stage, postal questionnaire was conducted that generated both qualitative and quantitative data. PARTICIPANTS: Of those contacted by telephone (164) from the United Kingdom, Australia, Canada, New Zealand and United States, 109 experts (66%) agreed to take part. Forty-three per cent (47/109) completed all three stages of the study. MEASUREMENTS: A Delphi technique was employed to gather data. The second and final questionnaires were constructed from the responses to the preceding questionnaires. Content analysis of the qualitative data was carried out at each stage. Statistical analyses of the influence of demographic factors, degree of shift in overall opinion between the first and second stages and degree of agreement between respondents at each stage were also conducted. FINDINGS: A consensus was reached on the strategies considered the most important and effective. Key areas include improving access to current information, improved staff training, addressing the issues of non pharmacy outlets and Internet pharmacy sites. Concerns were expressed regarding the possible conflict between commercial and customer interests. CONCLUSIONS: The consensus view presented offers practical and realistic guidance for policy makers and community pharmacists on the sale of OTC products. It reflects the best evidence to date of expert views in this area and accords with current UK guidelines. The effective implementation of these strategies can only be achieved with improved communication and coordination at local and national level. PMID- 12653819 TI - Major depression in patients with substance use disorders: relationship to 12 Step self-help involvement and substance use outcomes. AB - AIMS: Many patients treated for substance use disorders (SUDs) who become involved in 12-Step self-help groups have improved treatment outcomes. However, due to high rates of psychiatric comorbidity and major depressive disorder (MDD), among SUD patients in particular, concerns have been raised over whether these benefits extend to dual diagnosis patients. This study examined the influence of comorbid MDD among patients with SUDs on 12-Step self-help group involvement and its relation to treatment outcome. DESIGN: A quasi-experimental, prospective, intact group design was used with assessments completed during treatment, and 1 and 2 years postdischarge. PARTICIPANTS: A total of 2161 male patients recruited during in-patient SUD treatment, of whom 110 had a comorbid MDD diagnosis (SUD MDD) and 2051 were without psychiatric comorbidity (SUD-only). FINDINGS: SUD-MDD patients were initially less socially involved in and derived progressively less benefit from 12-Step groups over time compared to the SUD-only group. However, substance use outcomes did not differ by diagnostic cohort. In contrast, despite using substantially more professional out-patient services, the SUD-MDD cohort continued to suffer significant levels of depression. CONCLUSIONS: Treatment providers should allocate more resources to targeting depressive symptoms in SUD MDD patients. Furthermore, SUD-MDD patients may not assimilate as readily into, nor benefit as much from, traditional 12-Step self-help groups such as Alcoholics Anonymous, as psychiatrically non-comorbid patients. Newer, dual-diagnosis specific, self-help groups may be a better fit for these patients, but await further study. PMID- 12653820 TI - Linking alcohol- and drug-dependent adults to primary medical care: a randomized controlled trial of a multi-disciplinary health intervention in a detoxification unit. AB - AIM: Pragmatic approaches to integration of medical care and substance abuse treatment are desired. We assessed the effectiveness of a novel multi disciplinary clinic for linking patients in a residential detoxification program to primary medical care. PARTICIPANTS: We enrolled patients undergoing in-patient detoxification from alcohol, heroin or cocaine who had no primary care physician into a randomized controlled trial. The intervention consisted of a clinical evaluation at the detoxification unit in the health evaluation and linkage to primary care (HELP) clinic by a nurse, social worker and physician and facilitated referral to an off-site primary care clinic. The primary outcome of interest was attendance at a primary care appointment within 12 months. Secondary outcomes assessed over 24 months were addiction severity, health-related quality of life, utilization of medical and addiction services and HIV risk behaviors. FINDINGS: Of the 470 subjects enrolled, 235 were randomized to the HELP clinic intervention. Linkage to primary medical care occurred in 69% of the intervention group compared to 53% in the control group (P = 0.0003). The clinic was similarly effective for subjects with alcohol and illicit drug problems. Randomization to the HELP clinic resulted in no significant differences in secondary outcomes. CONCLUSIONS: The HELP clinic, a multi-disciplinary clinic located in a detoxification unit, effectively linked alcohol- and drug-dependent individuals to primary medical care. This intervention utilized a 'reachable moment', the period of addiction care, as a window of opportunity for linking substance abusers to medical care. PMID- 12653821 TI - Defining 'current drinkers' in national surveys: results of the 2000 National Alcohol Survey. AB - AIMS: The purpose of this study is to determine which respondents of national surveys who report low 'usual' past year drinking frequency, are unsure of their drinking frequency or refuse to answer, are misclassified either as 'current drinkers' or as 'ex-drinkers.' DESIGN AND SETTING: The data are from the 2000 National Alcohol Survey, a national household telephone probability sample of adults in all 50 US States and Washington, DC (n = 7612). PARTICIPANTS: A subsample of 1734 respondents who reported drinking 'less than once a month but at least once a year', 'less than once a year', 'refused' and 'don't knows' were also asked if they had consumed a whole drink of any alcoholic beverage in the 12 months using dates to anchor the question. MEASUREMENTS: Five alcohol measures were used to compare groups who changed or did not change their drinking status based on the followup question: volume, mean number of days 5 +, social consequences, dependence symptoms and DSM-IV. FINDINGS: The results indicate that only 4.6% of respondents (n = 349) changed their drinking status, and the percentages were equal in both directions. Women were more likely to change their drinking status and specifically become ex-drinkers based on the follow-up question. Respondents who became current drinkers drank significantly at a lower average volume than those who remained current drinkers. The prevalence rates based on the five alcohol measures were not affected by how 'current drinker' is defined. CONCLUSIONS: Adding the follow-up question is recommended for new alcohol surveys so that fewer respondents are misclassified. PMID- 12653822 TI - Efficacy of exercise counselling as an aid for smoking cessation: a randomized controlled trial. AB - AIM: To examine whether exercise counselling increases smoking abstinence and reduces tobacco withdrawal and gains in weight and body fat. DESIGN: A randomized controlled trial. SETTING: A community-based stop smoking clinic. PARTICIPANTS: Two hundred and ninety-nine male and female smokers. INTERVENTION: Participants were assigned randomly to a 7-week smoking cessation programme, including nicotine replacement therapy plus either (i) exercise counselling, or (ii) health education advice with equal contact time as for the exercise counselling condition. MEASUREMENTS: Six weeks of smoking abstinence was confirmed by expired carbon monoxide. FINDINGS: There was no significant difference in smoking abstinence between the exercise group (n = 154) and the controls (n = 145) at 6 weeks (39.6% versus 38.6%), nor was there any difference in gains in weight or body fat, although those in the exercise group increased their exercise levels. Exercise participants reported less tension, anxiety and stress than the controls during the first week of smoking abstinence (P = 0.03, 0.01 and 0.04, respectively), less irritability throughout 2 weeks of abstinence (P = 0.03), and less restlessness throughout 3 weeks of abstinence (P = 0.04). CONCLUSIONS: Adding brief exercise counselling to a smoking cessation programme did not increase smoking abstinence or reduce gains in weight or body fat significantly, although exercise levels were raised and there were some beneficial effects on psychological symptoms. PMID- 12653825 TI - Zero caffeine for kids. PMID- 12653828 TI - Iguana column. Trivializing health education. Portman Group style. PMID- 12653830 TI - Primary immunodeficiency diseases: an update. PMID- 12653829 TI - Immunological hurdles to lung gene therapy. AB - Gene delivery has the potential to offer effective treatment to patients with life-threatening lung diseases such as cystic fibrosis, alpha1-antitrypsin deficiency and lung cancer. Phase I/II clinical trials have shown that, in principle, gene transfer to the lung is feasible and safe. However, gene expression from both viral and non-viral gene delivery systems has been inefficient. In addition to extra- and intracellular barriers, the host innate and acquired immune system represents a major barrier to successful gene transfer to the lung. Results from studies in experimental animals and clinical trials have shown that inflammatory, antibody and T cell responses can limit transgene expression duration and readministration of the gene transfer vector. We will review here how the development of pharmacological and/or immunological agents can modulate the host immune system and the limitations of these strategies. A better understanding of the immunological barriers which exist in the lung might allow for a more sustained expression of the transgene and importantly help overcome the problem of readministration of viral vectors. PMID- 12653831 TI - Relevance of differential immunogenicity of human and mouse recombinant desmoglein-3 for the induction of acantholytic autoantibodies in mice. AB - Pemphigus vulgaris (PV) is an antibody-mediated autoimmune disease of the skin and mucous membranes. Desmoglein-3 (dsg-3) expressed in the suprabasal layer of the skin serves as an autoantigen in PV. Passive transfer of sera, either from patients with PV or from experimental animals immunized with a recombinant human dsg3 (hdsg3) into neonatal BALB/c mice results in blister formation, suggesting strongly that there is significant cross-reactivity between the mouse dsg3 (mdsg3) and the hdsg3. However, efforts to induce disease in adult mice through active immunization using hdsg-3 have not been successful, suggesting that the epitopes required for the induction of pathogenic antibodies in adult mice might not be present in hdsg3. Therefore, in this study, we expressed a full-length mdsg3 in insect cells and compared its serological reactivity with that of the hdsg3 using species specific polyclonal sera and a panel of seven monoclonal antibodies (MoAbs) with unique binding specificities to hdsg3. Studies using sera demonstrated a considerable cross-reactivity, while studies using MoAbs exhibited specific epitope differences between the two proteins. Because of these differences, we reasoned that immunization with mdsg3 might induce disease in adult mice. Immunization of four strains of mice (i.e. BALB/c, DBA/1, HRS/J and SJL/J) with mdsg3 resulted in considerable antibody response, but failed to induce lesions. However, sera from immunized BALB/c mice induced acantholysis of neonatal mouse skin in vitro. These studies indicated that our inability to induce lesions in adult mice through active immunization is not due to differences in the ability of mouse and human dsg3 to induce acantholytic antibodies, but due probably to structural differences between adult and neonatal mouse skin. Alternatively, immunization with a combination of dsg3 protein along with other proteins might be necessary to induce pemphigus disease in adult mice. Nevertheless, our current studies show that molecular mechanisms leading to the production of acantholytic antibodies in mice can now be studied using homologous mdsg3. PMID- 12653832 TI - Predicting death from tumour necrosis factor-alpha and interleukin-6 in 80-year old people. AB - Ageing is associated with low-grade inflammation and markers such as IL-6 possess prognostic value. Tumour necrosis-alpha (TNF-alpha) initiates the inflammatory cascade and has been linked to several age-associated disorders. It remains, however, unknown if TNF-alpha is associated with mortality in old populations. The aim of the present study was to investigate if serum levels of TNF-alpha were associated with all-cause mortality independently of interleukin (IL)-6 in a prospective study of 333 relatively healthy 80-year-old people. A Cox regression model was used to explore effects of TNF-alpha and IL-6 on survival in the following 6 years. A total of 133 participants died during this follow-up period. TNF-alpha was associated with mortality in men, but not in women, whereas low grade elevations in IL-6 were associated strongly with mortality in both sexes. TNF-alpha explained only 7% of the variability in IL-6 and effects of the two cytokines were independent of each other as well as of other traditional risk factors for death [smoking, blood pressure, physical exercise, total cholesterol, co-morbidity, body mass index (BMI) and intake of anti-inflammatory drugs]. These findings indicate that at least in old populations chronic elevated levels of TNF alpha and IL-6 have different biological functions that trigger age-associated pathology and cause mortality. PMID- 12653833 TI - Glomerular deposition of C1q and anti-C1q antibodies in mice following injection of antimouse C1q antibodies. AB - Anti-C1q autoantibodies are present in the serum of patients with different autoimmune diseases such as systemic lupus erythematosus (SLE). The occurrence of these autoantibodies correlates with renal involvement. In the present study we examined whether injection of rabbit antimouse C1q antibodies in mice leads to deposition in kidneys. Injection of healthy mice with a single dose of rabbit IgG antimouse C1q antibodies resulted in deposition of both C1q and IgG anti-C1q in glomeruli. The pattern of deposition observed in the glomeruli of mice injected with antimouse C1q antibodies both at 24 h and 2 weeks was both glomerular basement membrane (GBM)-associated and mesangial. Injection of control IgG did not have a detectable effect on circulating C1q levels, and no deposition of either C1q or rabbit IgG was seen at 24 h. The deposition of rabbit antimouse C1q and C1q in glomeruli resulted in complement activation, as assessed by C3 deposition, and influx of leucocytes associated with albuminuria in some, but not all mice. In none of the control mice was albuminuria observed. This report is the first to show that anti-C1q antibodies deposit in the healthy glomerulus together with autologous C1q. This deposition is stable for at least 2 weeks, causes complement activation, leucocyte influx and can lead to mild albuminuria. PMID- 12653834 TI - Augmented production of proinflammatory cytokines and accelerated allotransplantation reactions in heroin-treated mice. AB - Heroin treatment or abusive drug addiction influences many physiological functions, including the reactions of the immune system. Although suppression of various manifestations of the immune system after heroin (or morphine) administration has been reported, we show here that production of proinflammatory cytokines and nitric oxide (NO) was enhanced and allotransplantation reactions were accelerated significantly in heroin-treated recipients. Mice were treated by a subcutaneous administration of heroin (diacetylmorphine) given in one or repeated daily doses. The ability of spleen cells from treated mice to respond in vitro to alloantigens and to produce IL-2, IL-4, IL-10 and IFN-gamma, and the production of IL-1beta, IL-12 and NO by peritoneal macrophages, were tested. Within 2 h after heroin administration, proliferative responses to alloantigens and the production of IL-1beta, IFN-gamma, IL-12 and NO were enhanced significantly. In contrast, the production of anti-inflammatory cytokines IL-4 and IL-10 was at the same time rather decreased. As a consequence, skin allografts in heroin-treated mice were rejected more promptly than in untreated or vehicle-treated recipients. Similarly, the growth of allogeneic tumours induced by high doses of tumour cells was suppressed significantly in heroin treated mice. The enhancing effects of heroin on the production of proinflammatory cytokines were antagonized by naltrexone, a specific inhibitor of classic opioid receptors. These results show that heroin treatment augments production of proinflammatory cytokines and accelerates allotransplantation reactions. The observations thus illustrate the complexity of the effects of heroin on the immune system and should be taken into account during medical treatment of opiate addicts and in the use of morphine to decrease pain in various clinical situations. PMID- 12653835 TI - Differences in MHC-class I presented minor histocompatibility antigens extracted from normal and graft-versus-host disease (GVHD) mice. AB - Graft-versus-host disease (GVHD) may develop after allogeneic bone marrow transplantation (BMT) between donors and recipients incompatible for minor histocompatibility antigens (mHAg). Here, we examined the possible relationship between tissue-specific distribution of dominant mHAg peptides and specific organ destruction caused by GVHD. In the B6 anti-Balb/b (H-2b) strain combination, a GVHD developed against Balb/b mHAgs. Despite the high number of incompatible mHAgs between these two strains, both cytotoxic T lymphocyte (CTL) response and GVHD could be attributed to a limited number of dominant mHAgs. We studied CTL defined expression of dominant mHAgs in normal tissues and their GVHD-related modifications. mHAg peptides were prepared by acid elution and reversed-phase high pressure liquid chromatography fractionation from the spleen, liver, gut and skin as GVHD target tissues and from the heart and kidney as control tissues. Peptidic fractions extracted from normal and GVHD tissues were incubated with RMA S targets and analysed using bulk B6 anti-Balb/b CTL. In each tissue several fractions were recognized with a given pattern of mHAg expression. GVHD induced qualitative and quantitative changes in antigenic peptide expression. Modifications in mHAg presentation during GVHD concerned preferentially GVHD target organs as opposed to non-GVHD target organs. In addition, when immunizing tissues were derived from GVHD mice instead of normal mice, the profile of CTL recognition was different. In conclusion, these data indicate that broad differences could exist in peptide presentation between various normal and GVHD target organs. PMID- 12653836 TI - Calcium signalling is altered in myeloid cells with a deficiency in NADPH oxidase activity. AB - The relation of O(2.-)-production and Ca2+ homeostasis was investigated in PLB 985 cell lines and neutrophilic granulocytes from peripheral blood. In differentiated wild-type PLB-985 cells, a high level of O(2.-)-production was associated with a significant decrease in the membrane potential and the inhibition of capacitative Ca2+ entry. These correlations were not observed in gp91phox -/- cells or in cells transfected with a non-functional mutant of gp91phox (Thr341Lys). Membrane depolarization and inhibition of Ca2+ entry reappeared in cells transfected with wild-type gp91phox. These experiments demonstrate that inhibition of Ca2+ entry depends on the presence of a functional NADPH oxidase. The Ca2+ signal induced by stimulation of chemotactic receptors also showed remarkable differences: [Ca2+]ic in the sustained phase was higher in gp91phox-/- than in wild-type cells. Alteration of the Ca2+ signal was reproduced by treating peripheral blood neutrophils with the NADPH oxidase inhibitor diphenylene-iodonium. It is concluded that the deficiency in O(2.-)-production is accompanied by significant alterations of Ca2+ homeostasis in myeloid cells. PMID- 12653838 TI - CpG ODN enhances uptake of bacteria by mouse macrophages. AB - Unmethylated CpG motif in synthetic oligodeoxynucleotide (CpG ODN) or bacterial DNA is well recognized for its role in innate immunity, including enhancing production of NO and cytokines by macrophages. In the present study, we demonstrated the effect of CpG ODN on the phagocytic uptake of bacteria by macrophages. Flow cytometric analysis of mouse macrophages (RAW 264.7) incubated with fluorescein isothiocyanate (FITC)-labelled Burkholderia pseudomallei, Salmonella enterica serovar Typhi or Escherichia coli showed that CpG ODN increased the uptake of these bacteria by mouse macrophages. The enhancement of bacterial uptake by CpG ODN was concentration-dependent. The increase of bacterial uptake by CpG ODN-activated macrophages shown above is consistent with the result of bacteria internalization study using a standard antibiotic protection assay. There was also an increase in the rate and degree of multi nucleated giant cell formation, phenomena which have been shown previously to be unique when the cells were infected with B. pseudomallei. These observations may provide significant insights for future investigation into host cell-pathogen interaction. PMID- 12653837 TI - Keratinocyte growth factor (KGF) decreases ICAM-1 and VCAM-1 cell expression on bronchial epithelial cells. AB - Activation of leucocytes during airway inflammatory reaction involves adhesion to bronchial epithelial cells (BEC), a process implicating specific interactions between glycoproteins with epithelial cell surface proteins, mainly intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). In this study, the effect of keratinocyte growth factor (KGF), a growth factor involved in pulmonary epithelium repair, was evaluated on adhesion molecule expression with BEAS-2B cells and BEC and granulocyte adherence to BEAS-2B. The modulation by KGF of membrane and mRNA expression of ICAM-1 and VCAM-1 was studied on confluent cells stimulated or not with tumour necrosis factor-alpha (TNF) (200 UI/ml) or TNF and interleukin (IL)-4 (50 UI/ml and 10 ng/ml). Levels of soluble-(s)ICAM-1 and sVCAM-1 were measured by ELISA. Although moderately, KGF significantly decreased membrane ICAM-1 expression in unstimulated BEAS-2B cells (24% inhibition at 100 ng/ml) or in TNF- or TNF + IL-4-stimulated cells (22.5 and 18.7% inhibition, respectively). Treatment with KGF tended to decrease VCAM-1 expression in TNF- and TNF + IL-4-stimulated BEAS-2B (P = n.s. and P < 0.05, 14 and 15% inhibition, respectively). In primary culture of BEC, adhesion molecule expression was also reduced. ICAM-1 and VCAM-1 mRNA expression were also inhibited by KGF. Levels of sICAM-1 and sVCAM-1 were not significantly increased in supernatants from KGF-treated cells (30% and 24% increase at 100 ng/ml, respectively) compared to controls. Moreover, KGF decreased by 31% the adherence of neutrophils to TNF-activated BEAS-2B. In conclusion, KGF decreases ICAM-1 and VCAM-1 expression and neutrophil adherence in BEC. These suggest its involvement in the resolution of the inflammatory reaction. PMID- 12653839 TI - Evaluation of CD4+ T cells proliferating to grass pollen in seasonal allergic subjects by flow cytometry. AB - Our objective was to characterize T-cell responses to Phleum pratense in grass pollen allergic individuals and healthy controls using the fluorescent dye PKH26. Peripheral blood mononuclear cells were stimulated with P. pratense, or with recall antigens, and CD3+/CD4+ and CD3+/CD8+ T-cells that had proliferated were analysed by flow cytometry. In the presence of P. pratense CD4+/CD3+ T-cells proliferated more in grass pollen sensitive atopic patients than in nonallergic controls or in nongrass pollen sensitive atopic subjects. PPD and TT recall antigens elicited uniformly high proliferation in all T-cell subsets. Only half of pollen sensitive patients also had an increased proliferation of CD3+/CD8+ T cells in response to P. pratense. We determined precursor frequency of CD4+ T cells in the original population that responded to P. pratense and found values ranging from 1 x 10-3 to 0.6 x 10-1, in the same range as those measured for PPD and TT. In conclusion, grass pollen sensitive atopic patients show enhanced CD4+ T-cell reactivity to P. pratense, and this could be related to the presence of elevated numbers of circulating allergen-specific CD4+ T cells. This flow cytometric method should allow the identification of other phenotypic markers such as intracellular cytokines in allergen specific responding CD4+ T cells. PMID- 12653841 TI - Restricted genetic defects underlie human complement C6 deficiency. AB - Complement C6 homozygous deficiency (C6D) has been rarely observed in Caucasians but was reported at higher prevalence among African-Americans. We report on the molecular basis of C6D in seven unrelated black individuals of North or Central Africa descent who live in France. These patients have presented Neisseria meningitidis infection (four cases), focal and segmental glomerulosclerosis with hyalinosis (one case), systemic lupus erythematosus (one case) or Still's disease (one case). All patients exhibited undetectable antigenic C6 by using a sensitive ELISA assay. An additional four cases of complete C6 deficiency with no associated disease have been characterized after family studies. Exons 6, 7 and 12 have been described recently as the location of molecular defects on the C6 gene in randomly chosen black Americans. Genomic DNA from the seven patients were subjected to direct polymerase chain reaction amplification of these three exons. Nucleotide sequencing analysis of the amplified DNA fragments revealed a homozygous single-base deletion (1936delG) in exon 12 in three cases and four compound heterozygous deletions for a single base in exon 7 (1195delC) or in exon 6 (878delA) associated with the same deletion in exon 12 (1936delG). Our observations further establish the restricted pattern of genetic defects associated with homozygous C6 complement deficiency in individuals of African descent. PMID- 12653840 TI - Clearance of red cells by monoclonal IgG3 anti-D in vivo is affected by the VF polymorphism of Fcgamma RIIIa (CD16). AB - Human red cells (RBC) coated with IgG anti-D are cleared from the circulation to the spleen by macrophages which express IgG receptors (Fcgamma R). Polymorphisms of Fcgamma RIIa and Fcgamma RIIIa affect IgG binding in vitro, and may alter the efficiency of clearance of immune complexes in vivo. In a RBC clearance study, 22 Rh D-negative subjects were given 100-400 micro g human monoclonal or polyclonal IgG anti-D i.m. followed 48 h later by 51Cr-labelled D+ RBC. The half lives of the infused D+ RBC were determined, together with the coating levels of anti-D on the D+ RBC. Fcgamma RIIA and FcgammaRIIIA genotyping was performed. Large ranges of phagocytosis and extracellular lysis of RBC in vitro, and of half lives of RBC in vivo, were observed. Clearance of RBC coated with monoclonal IgG3 anti-D (BRAD 3) was more rapid in five subjects homozygous for Fcgamma RIIIa-F/F158 than in three subjects expressing the Fcgamma RIIIa-V158 allele (P = 0.024). This effect was not observed, however, for those individuals given polyclonal anti-D. There was also no significant difference in the efficiency of RBC destruction in vitro or of RBC clearance in vivo between the subjects analysed for individual genotypes or alleles or combinations of alleles. In conclusion, the presence of the Fcgamma RIIIa-V158 allele compromised the efficiency of removal of RBC coated with IgG3 anti-D. PMID- 12653842 TI - Mannan-binding lectin and hepatitis C infection. AB - Japanese patients with chronic hepatitis C infection unresponsive to treatment with interferon possessed genotypes disproportionately conferring low mannan binding lectin (MBL) concentrations. Our aims were to confirm or refute this finding in European patients at the MBL protein level, and to investigate whether a low circulating concentration of MBL might influence susceptibility to, or disease progression from, hepatitis C viral infection. Serum samples obtained from 180 hepatitis C patients and 566 blood donors were assayed for MBL. MBL concentrations were related to disease characteristics retrieved from patients' records. MBL concentrations were higher in hepatitis C patients (median 2.5 microg/ml versus 1.3; P < 0.0001) and the proportion of patients with very low (MBL-deficient) concentrations was similar to that of the healthy controls. There were no significant associations between patients with low serum MBL and the disease features studied, including response to antiviral therapy. Therefore, low circulating MBL does not increase susceptibility to hepatitis C infection, and MBL concentration does not have a major influence on the course of the disease or the response to antiviral therapy. MBL replacement therapy would therefore not be indicated for chronic hepatitis C patients who failed to respond fully to treatment with interferon and ribavirin. PMID- 12653843 TI - Cellular responses to cytomegalovirus in immunosuppressed patients: circulating CD8+ T cells recognizing CMVpp65 are present but display functional impairment. AB - The availability of tetrameric complexes of HLA class I molecules folded with immunodominant peptides makes it possible to utilize flow cytometry for rapid and highly specific visualization of virus specific CD8+ T cells. An alternate technique is to incubate whole blood with specific antigens and to subsequently detect and characterize responding T cells (e.g. by performing intracellular staining of interferon-gamma). By using an HLA-A2 tetramer construct folded with the same immunodominant CMV-peptide as that used for peptide pulsing, we monitored both the presence and functional capacity of CMV-specific CD8+ T cells. In addition T cell activation was assayed by determination of CD38 and CD69 expression. Twelve organ transplant patients and 31 healthy blood donors with latent CMV infection were investigated using CMV pp65 tetramer staining and intracellular staining of interferon-gamma after CMV pp65 peptide pulsing or CMV lysate pulsing. CMV-specific T cells were detected in similar absolute numbers as well as frequencies of T cells in the two groups investigated. However, the CMV specific CD8+ T cells in immunosuppressed individuals showed a decreased functional response to the CMV-peptide, as evidenced by reduced interferon-gamma production when compared to healthy blood donors (19%; 42%, P < 0.005). In addition, CD38 expression was markedly higher in immunosuppressed patients compared to healthy blood donors (24%; 6%, P < 0.005). In a case report we demonstrate that reactivation of CMV can occur in an immunosuppressed patient with high number of CMV-specific T cells, but without functional capacity. Hence, these findings reflect impaired activation of cytotoxic T cells controlling latent CMV infection in immunosuppressed patients. PMID- 12653844 TI - Preserved immune system in long-term asymptomatic vertically HIV-1 infected children. AB - The objective of this study was to study immune system status in long-term asymptomatic (LTA) HIV-1-infected children. A cross-sectional study was used, involving HIV-1-infected children over 7 years of age who were rated into two groups according to their clinical and immunological classification: (a) LTA: 7 asymptomatic HIV-1-infected children in A1; (b) Rapid progressor (RP): 14 age matched C3 HIV-1-infected children. The control group consisted of 17 age-matched uninfected children. The characterization of CD4+ T-cell subsets was determined by three-colour flow cytometry. The proliferative response and cytokine production by activated peripheral blood T-cells were also measured. IL-7 levels were measured in serum. Thymic production of T-cells was quantified by TCR rearrangement excision circles (TRECs). The LTA children showed similar proliferative responses to PHA, PWM and anti-CD3+ anti-CD28, but lower responses to tetanus toxoid and streptokinase, in comparison with the controls but always higher responses in comparison with the RP group. The production of TNF-alpha and IFN-gamma was similar in the LTA and control groups, and both were higher than the levels in the RP group. The LTA group showed a lower percentage of memory CD4+ T-cells (CD4+ CD45RO+, CD4+ CD45RA-CD62L+) than the control and RP groups. The LTA group also showed lower percentages of CD4+ CD7- cells than the controls. As for naive CD4+ T-cells (CD4+ CD45RA+ CD62L+), CD4+ CD45RA+ and CD4+ CD62L+ cells, the LTA group showed higher values than the control and RP groups. The LTA group showed higher percentages of CD4+ HLA-DR+ CD38+ than the controls, but lower values than the RP group. In contrast, the LTA group had percentages of CD4+ HLA-DR-CD38+ T-cells higher than both the control and RP groups, whereas CD4+ CD38+ levels were only higher in the LTA group in comparison with the controls. CD4+ HLA-DR+ CD38- and CD4+ HLA-DR+ cell numbers were lower in the LTA group in comparison with the RP group. We found almost normal values of TRECs and IL-7 in the LTA group, but lower values in the RP group. Moreover, we found an inverse relation between TREC levels and IL-7 in plasma from HIV-infected children. Asymptomatic HIV-1 infected children have a well preserved immune system similar to that of control uninfected children in spite of HIV-infection for more than 7 years. Moreover, our results identified new markers of HIV disease, such as TRECs and IL-7, that could be used to monitor disease. PMID- 12653845 TI - Role of incidental and/or cured intestinal parasitic infections on profile of CD4+ and CD8+ T cell subsets and activation status in HIV-1 infected and uninfected adult Ethiopians. AB - Intestinal parasitic infections have been suggested to cause persistent immune activation leading to an unbalanced immune state. Such a state has been proposed to be a major factor in the pathogenesis of AIDS in an African context. The present study investigated the effect of incidental parasitic infection and treatment on the profile of T cell differentiation and activation markers on CD4+ and CD8+ T cells from HIV-1 infected and uninfected adult Ethiopians. Cryopreserved PBMCs from 64 subjects (41 HIV-negative and 23 HIV-positive) with follow-up visits at 6-monthly intervals were used to compare the effect of incidental intestinal parasites and their treatment upon T cell subset profiles and activation status. The samples were stained with antibodies to various T cell differentiation and activation markers allowing naive, memory, effector, memory/effector, activated and resting CD4+ and CD8+ T cell subsets to be quantified by triple-colour FACScan. Incidental intestinal parasitic infections resulted in a significant increase in memory CD4+ T cell numbers both in HIV negative and HIV-positive subjects (P < 0.05). There was also a significant increase in the percentage of CD8+ HLA-DR+ T cells (P < 0.05) in HIV-positive subjects co-infected with parasites. In HIV-negative subjects, a significant decline in activated cells and a significant increase in resting CD8+ T cells (P < 0.05) was observed after treatment for parasites. These data suggest that intestinal parasitic infections could result in the alteration of T cell subset counts and also in the up-regulation of T cell activation markers in peripheral blood. Treatment of parasitic infections showed a tendency to reduce the activation suggesting that, together with other community based intervention strategies, such treatment could be used to down-regulate immune activation and hence protect the host from being easily attacked by HIV. PMID- 12653846 TI - Virus load correlates inversely with the expression of cytotoxic T lymphocyte activation markers in HIV-1-infected/AIDS patients showing MHC-unrestricted CTL mediated lysis. AB - Cytotoxic T lymphocytes (CTL) are key players to suppress viral load (VL) but CTL responses become compromised with progression of HIV-infection/AIDS. Some progressors develop MHC-unrestricted CTL with anti-CD4+ cytocidal activity. Immune activation status of these CTL and its significance in disease progression are unknown. To determine the relationship between VL and T cell activation, a cross-sectional study was carried out using blood samples from 13 HIV-1 infected/AIDS patients at various stages of progression and seven age-matched seronegative controls. We examined expression of HLA-DR and CD38 activation markers on purified CTL. MHC-unrestricted killing by these CTL was also evaluated against uninfected, allogeneic CD4+ T cells as well as several human cell lines. The expression of activation markers correlated inversely (rs = - 0.91, P < 0.0001) with VL of the subjects. CTL effectors of these patients killed targets expressing or lacking CD4+, independently of MHC class I recognition. Interestingly, the patients with higher VL showed an increased number of gammadeltaTCR-bearing CTL in blood and their MHC-unrestricted killing activity was blocked significantly (P < 0.01) by gammadeltaTCR-specific monoclonal antibody. CD3+ T counts of these patients were also consistently subnormal. Inverse correlation between VL and CD8+ T cell activation markers seems to be an indicator of CTL-associated immunopathogenesis in HIV patients with elevated gammadeltaCTL in the peripheral blood. PMID- 12653847 TI - Anti-cytokine autoantibodies in autoimmunity: preponderance of neutralizing autoantibodies against interferon-alpha, interferon-omega and interleukin-12 in patients with thymoma and/or myasthenia gravis. AB - We have screened for spontaneous anticytokine autoantibodies in patients with infections, neoplasms and autoimmune diseases, because of their increasingly reported co-occurrence. We tested for both binding and neutralizing autoantibodies to a range of human cytokines, including interleukin-1alpha (IL 1alpha), IL-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-18, interferon-alpha2 (IFN-alpha2), IFN-omega, IFN-beta, IFN-gamma, tumour necrosis factor alpha (TNF alpha), transforming growth factor beta-1 (TGF-beta1) and granulocyte-macrophage colony stimulating factor (GM-CSF), in plasmas or sera. With two notable exceptions described below, we found only occasional, mostly low-titre, non neutralizing antibodies, mainly to GM-CSF; also to IL-10 in pemphigoid. Strikingly, however, high-titre, mainly IgG, autoantibodies to IFN-alpha2, IFN omega and IL-12 were common at diagnosis in patients with late-onset myasthenia gravis (LOMG+), thymoma (T) but no MG (TMG-) and especially with both thymoma and MG together (TMG+). The antibodies recognized other closely related type I IFN alpha subtypes, but rarely the distantly related type I IFN-beta, and never (detectably) the unrelated type II IFN-gamma. Antibodies to IL-12 showed a similar distribution to those against IFN-alpha2, although prevalences were slightly lower; correlations between individual titres against each were so modest that they appear to be entirely different specificities. Neither showed any obvious correlations with clinical parameters including thymoma histology and HLA type, but they did increase sharply if the tumours recurred. These antibodies neutralized their respective cytokine in bioassays in vitro; although they persisted for years severe infections were surprisingly uncommon, despite the immunosuppressive therapy also used in most cases. These findings must hold valuable clues to autoimmunizing mechanisms in paraneoplastic autoimmunity. PMID- 12653848 TI - Altered CD8+ T cell responses to selected Epstein-Barr virus immunodominant epitopes in patients with multiple sclerosis. AB - An increased frequency of antiviral CD8+ T cells is seen in chronic viral infections. During herpes virus infections the expanded CD8+ T cells are thought to control the reactivation of the latent infection. Because multiple sclerosis (MS), a presumed autoimmune disease of the central nervous system, has been associated with a late Epstein-Barr virus (EBV) infection, we wished to examine whether the CD8+ T cell response to EBV epitopes differed between MS patients and healthy controls. Here we report an increased frequency of CD8+ T cells responding to EBV epitopes from nuclear antigen 3 A (HLA-A2/CLG) and latent membrane protein 2 (HLA-B7/RPP) in MS patients. Noticeably, the altered CD8+ T cell response occurred to some but not all EBV epitopes and did not reach the high level seen during acute infection. The responses towards two immunodominant epitopes from human cytomegalovirus (HCMV) were similar in MS patients and normal controls. Together, our data demonstrate the presence of an increased frequency of CD8+ T cells reacting with two epitopes from EBV in patients with MS. The altered response to only two of the tested EBV epitopes would be consistent with the presence of cross-reactive epitopes. PMID- 12653849 TI - Immunoglobulins in children with epilepsy: the Dutch Study of Epilepsy in Childhood. AB - In an unselected cohort of 282 children, serum immunoglobulin (Ig) concentrations were determined shortly after the first presentation with one or more unprovoked epileptic seizures and before the start of treatment with anti-epileptic drugs (AEDs), and after 9-18 months of AEDs use. At intake, IgA, IgG1, IgG2 and IgG4 concentrations were significantly higher than published reference values in healthy age-matched controls. In a subset of 127 children, Ig levels at intake were compared with those after AEDs use for 9-18 months. IgA and IgG4 levels had decreased significantly to normal concentrations, but IgG1 and IgG3 levels increased significantly. To determine the influence of AEDs, Ig levels in children who used carbamazepine or valproic acid monotherapy were analysed separately. The use of carbamazepine was associated with a significant decrease of IgA and IgG4 levels, and the use of valproic acid with a significant decrease of IgA and increase of IgG1 levels. In conclusion, humoral immunity is already altered in children shortly after the first presentation with epileptic seizures. Whether this is the consequence of an exogenous event, and to what extent this is related to an interaction of the central nervous system and the immune system, remains to be evaluated. Treatment with AEDs, such as carbamazepine and valproic acid, is associated with significant changes of Ig (sub)class concentrations. PMID- 12653850 TI - Increased circulating interleukin-12 (IL-12) p40 in pulmonary sarcoidosis. AB - In sarcoidosis, a T helper 1 (Th1) response is an essential event and the up regulation of interleukin-12 (IL-12) has been detected in affected disease sites. In order to investigate the clinical usefulness of circulating IL-12, we measured the serum concentrations of IL-12 by ELISA and performed immunohistochemistry using specific MoAbs for IL-12 in the lungs and scalene lymph nodes of patients with sarcoidosis. The serum concentration of IL-12 p40 was detectable in all 45 patients with pulmonary sarcoidosis and 18 normal controls, whereas that of IL-12 p70 was undetectable. The serum concentrations of IL-12 p40 in pulmonary sarcoidosis were significantly higher than those of the normal controls, especially in cases with abnormal intrathoracic findings detected by chest roentogenogram. The serum concentrations of interferon-gamma (IFN-gamma) also increased compared with those of normal controls and there was a significant positive correlation between the serum concentrations of IL-12 p40 and IFN-gamma. Furthermore, serum angiotensin-converting enzyme (ACE) and lysozyme, which are known to be useful markers for disease activity in sarcoidosis, correlated well with the serum concentrations of IL-12 p40. The positive 67Ga scan group (for lung field) had significantly elevated serum IL-12 p40 levels compared with those of the negative group. No bioactivity of IL-12 p70 was detected in three sarcoid cases sera by using the IL-12 responsive cell line. Finally, the immunohistochemical approach revealed that IL-12 p40 was expressed in the epithelioid cells and macrophages of sarcoid lungs and lymph nodes. We concluded that the production of IL-12 p40 was far greater in the sera and we have demonstrated this to be a useful clinical marker for disease activity and the Th1 response in pulmonary sarcoidosis. PMID- 12653851 TI - High serum vascular endothelial growth factor correlates with disease activity of spondylarthropathies. AB - Angiogenesis is involved in chronic inflammatory joint diseases such as rheumatoid arthritis (RA). Vascular endothelial growth factor (VEGF) plays a crucial role in angiogenesis. The spondylarthropathies (SpA) are characterized by enthesitis and synovitis, in which blood vessels participate. The objective of this study was to investigate serum VEGF levels and their potential associations with disease activity markers for SpA. Sera were collected from 105 patients with SpA (72 with ankylosing spondylitis (AS), four with psoriatic arthritis (PsA), six with reactive arthritis (ReA), eight with enteropathic arthropathy and 15 with undifferentiated SpA), 50 patients with rheumatoid arthritis (RA) and 64 healthy controls. Disease activity in SpA patients was assessed using the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and laboratory parameters of inflammation [erythrocyte sedimentation rate (ESR) and C-reactive protein level (CRP)]. Serum VEGF levels were significantly higher in SpA patients (316.4 +/- 215.6 pg/ml) and RA patients (405.2 +/- 366.5) than in controls (217.3 +/- 145.2) (P = 0.003). In SpA patients, serum VEGF levels correlated with disease activity indices (BASDAI: r = 0.22, P = 0.04; ESR: r = 0.3, P = 0.003; and CRP: r = 0.23, P = 0.02). Serum VEGF levels were not associated with presence of extra articular manifestations or syndesmophytes or with the grade of sacroiliitis. These results suggest that VEGF and therefore angiogenesis may play a role in SpA pathogenesis and may serve as a disease activity marker in SpAs. PMID- 12653852 TI - Elevated angiogenin levels in synovial fluid from patients with inflammatory arthritis and secretion of angiogenin by cultured synovial fibroblasts. AB - Angiogenesis is a key process in the pathogenesis of inflammatory arthritis. Angiogenin is one of the most potent inducers of neovascularization in experimental models in vivo. To look for evidence that angiogenin is involved in inflammatory joint disease, we examined plasma and synovial fluid (SF) samples from rheumatology patients and synovial fibroblast cell culture supernatants. Angiogenin levels were determined by radioimmunoassay and ELISA. Plasma angiogenin concentrations ranged from 96 to 478 ng/ml, with no significant difference between patients and normal controls. In SF, angiogenin concentrations were significantly higher in patients with acute or chronic synovitis (rheumatoid arthritis (RA): median, 104 ng/ml; range 13-748, n = 14; crystal-induced arthritis (CIA): median, 149 ng/ml; range, 37-616, n = 14, and other chronic inflammatory arthritis: median, 42 ng/ml; range, 15-205; n = 9) than in the 18 patients with osteoarthritis (OA) (median, 20 ng/ml; range 8-116) (P < 0.0001, anova). Angiogenin levels in SF from RA patients in remission with secondary OA were similar to those achieved in primary OA, and decreased in parallel with the resolution of acute gout. Angiogenin protein was released by cultured synovial fibroblasts from OA and RA patients, and reached 1.18 ng/106 cells/day. These data suggest that angiogenin may mediate local inflammation in arthritis via effects on angiogenesis and leucocyte regulation. PMID- 12653853 TI - CTLA-4 (CD152) expression in peripheral blood T cells in Kawasaki disease. AB - Kawasaki disease (KD) is an acute febrile illness of early childhood caused by vasculitis. Whether or not peripheral blood T cells are activated in acute KD remains uncertain, as some reports have presented evidence of peripheral blood T cell activation, whereas others suggest that the level of peripheral blood T cell activation is low during acute KD. Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4, CD152) is a surface molecule of activated T cells. We therefore investigated intracellular CTLA-4 expression in the peripheral blood T cells of patients with acute KD as a marker of T cell activation. We collected blood samples from 20 patients with KD and six with Epstein-Barr virus infectious mononucleosis (EBV-IM) who were admitted to our hospital, as well as 13 healthy children. We determined the intracellular expression of CTLA-4 in T cells by flow cytometry. We demonstrated that the intracellular expression of CTLA-4 is up regulated in peripheral blood CD3+ T cells, CD4+ T cells and CD8+ T cells at the early part of the acute stage in KD. However, the mean percentages of intracellular T cells expressing CTLA-4 in EBV-IM patients were about fourfold higher than those in T cells from patients with acute KD. Our results suggested that the level of activation of peripheral blood T cells is very low during acute KD. PMID- 12653854 TI - Production of antineutrophil cytoplasm antibodies derived from circulating B cells in patients with systemic vasculitis. AB - The pathogenesis of systemic vasculitis is complex and is likely to involve many mechanisms. In certain systemic vasculitides, autoimmunity plays an important role with autoantibodies developing towards neutrophils, which are termed antineutrophil cytoplasm antibodies (ANCA). There is a growing body of evidence that T cells may contribute to the pathogenesis of ANCA-associated vasculitides. A system was set up to determine whether B cells require T cell help to produce antibodies in a peripheral blood lymphocyte (PBL) culture system enriched for B cells and dendritic cells (DC). As a control, tetanus toxoid (TT) antibody production was detected from individuals not recently immunized with tetanus vaccine when stimulated with TT antigen. Proteinase 3 (PR3) and myeloperoxidase (MPO) antibodies were produced from B cell and DC enriched cultures prior to the addition of antigen in some ANCA-positive patients with high ANCA titres, but not from patients with low ANCA titres or controls. PBMC from individuals recently immunized with tetanus vaccine were also maximally stimulated in that addition of antibody did not enhance antibody production. We conclude that this system supports a role for T cell help in the production of TT antibodies in individuals not immunized recently with tetanus vaccine. However, in patients with ANCA associated vasculitis and controls recently immunized with tetanus vaccine, circulating B cells are apparently spontaneously producing autoantibody, possibly reflecting a system already maximally driven in vivo, and therefore masking underlying potential T cell-B cell collaboration. Such B cells may be less responsive to regulatory stimuli in vivo. PMID- 12653856 TI - Cyclic guanosine monophosphate phosphodiesterase activity in human gingival carcinoma. AB - BACKGROUND: Cyclic guanosine monophosphate (cGMP) is an essential second messenger metabolized by phosphodiesterases (PDEs). OBJECTIVES: We looked for a possible correlation of PDE activities in human oral squamous cell carcinoma (OSCC) with and without lymph node metastases. MATERIALS AND METHODS: The analysis of phosphodiesterase activity and the cGMP assay were done by reverse phase HPLC on samples of fresh or frozen gingival tissues. Analysis of cGMP was confirmed with the enzyme-linked immunoabsorption assay. RESULTS AND CONCLUSIONS: cGMP PDE activity was 34.92 +/- 7.17 SD, 12.89 +/- 4.43 SD, and 35.88 +/- 8.76 SD (nmols/mg of protein), respectively, in controls, samples without lymph node involvement (N-), and specimens with lymph node metastases (N+). cGMP values were 1.97 +/- 0.63 SD, 3.30 +/- 1.47 SD, and 3.49 +/- 1.47 SD (nmols/mg of protein). Our data support the hypothesis of a role for cGMP and PDE in the progression of OSCC. PMID- 12653857 TI - Increase of mast cells and tumor angiogenesis in oral squamous cell carcinoma. AB - BACKGROUND: Although mast cells (MCs) have been implicated in promoting angiogenesis in some malignant tumors, especially of the aerodigestive tract, little is known in oral squamous cell carcinoma (SCC). METHODS: A retrospective study was conducted to elaborate upon the correlation between MCs and tumor angiogenesis in 26 cases of oral SCC, six cases of oral pre-malignant dysplasia, 10 cases of oral hyperkeratosis, and six cases of normal oral mucosa by means of immunohistochemical technique. RESULTS: The MCs in all lesions and normal oral mucosa strongly expressed tryptase. The densities of MCs and microvessels appeared to increase with disease progression. The MC and microvascular counts were significantly higher in oral SCC than in hyperkeratosis and normal oral mucosa (P < 0.05). A significant correlation between MC and microvascular densities was observed in oral SCC (r = 0.5; P = 0.012). CONCLUSIONS: These findings suggest that MCs may upregulate tumor angiogenesis in oral SCC, perhaps via MC tryptase. PMID- 12653858 TI - The multiform and variable patterns of onset of orofacial granulomatosis. AB - BACKGROUND: The recurrent chronic orofacial swelling caused by orofacial granulomatosis (OFG) can cause significant cosmetic and functional problems but can be prevented if the disease is diagnosed early and promptly treated. Although the enlargement of the lips is described to be the most common presenting complaint, the clinical onset of OFG may be characterized by minor associated mucosal and neurological manifestations, making early diagnosis very difficult or, sometimes, merely presumable. PATIENTS AND METHODS: We retrospectively analyzed the clinical manifestations of 19 patients with OFG, who were examined at our institution between 1998 and 2002, in order to determine their initial manifestations and presenting symptoms. RESULTS: A total of 10 patients showed classical recurrent enlargement of the lips (six lower; four upper) as presenting symptom. In the other nine patients, OFG onset was characterized by transient unilateral facial nerve palsy (two cases), intraoral manifestations (two cases), recurrent swelling of the periorbital area (two cases), of the chin (one case), of the zygomatic area (one case), and of the cheeks (one case). CONCLUSION: Our data underlined that OFG onset could be frequently characterized by widely variable, multiform, and temporary clinical findings. Involvement of atypical sites of the orofacial region and presence of single minor manifestations may occur as presenting symptoms, often preceding the development of traditional clinical findings. PMID- 12653859 TI - Levamisole and Chinese medicinal herbs can modulate the serum interleukin-6 level in patients with recurrent aphthous ulcerations. AB - BACKGROUND: Recurrent aphthous ulcerations (RAU) are common oral inflammatory lesions. Interleukin-6 (IL-6) is a pro-inflammatory cytokine that has effects on cellular and humoral immunities. Previous studies have shown that the high serum IL-6 levels in some RAU patients can be reduced by drug treatment. This finding suggests that IL-6 may be a useful marker in evaluating therapeutic effects of RAU. METHODS: In this study, we used a solid phase, two-site sequential chemiluminescent immunometric assay to determine the baseline serum levels of IL 6 in a group of 228 patients with RAU, erythema multiforme (EM), traumatic ulcers (TU), oral submucous fibrosis (OSF), pemphigus vulgaris (PV), or Sjogren's syndrome (SS), and in 77 normal control subjects. Some RAU patients were treated with levamisole plus Chinese medicinal herbs or levamisole only for 0.5-5 months and their serum IL-6 levels were measured after treatment. RESULTS: We found that about 99% of the normal control subjects and the patients with EM, TU, or OSF had a serum IL-6 level within the normal limit of 5.0 pg/ml. However, 24% (48/197) RAU patients, 14% (1/7) EM patients, 43% (3/7) PV patients, and 100% (6/6) SS patients had a serum level of IL-6 greater than 5.0 pg/ml. The mean serum level of IL-6 in patients with RAU (3.6 +/- 3.5 pg/ml, P < 0.001), minor type RAU (2.7 +/- 2.0 pg/ml, P < 0.05), major type RAU (5.2 +/- 4.6 pg/ml, P < 0.001), or herpetiform type RAU (4.1 +/- 3.8 pg/ml, P < 0.01) was higher than that in normal control subjects. The mean serum level of IL-6 in major type (P < 0.001) or in herpetiform type RAU patients (P < 0.05) was higher than that in minor type RAU patients. The mean reduction of serum IL-6 level (10.0 +/- 7.1 pg/ml) in RAU patients after treatment with levamisole plus Chinese medicinal herbs was significantly higher than that (5.1 +/- 3.7 pg/ml) in RAU patients after treatment with levamisole only (P < 0.005), suggesting that the combination therapy is superior to the single therapy of levamisole only. CONCLUSION: We conclude that levamisole and levamisole plus Chinese medicinal herbs can modulate the serum IL-6 level in RAU patients. Although the therapeutic effect of RAU can be assessed by a decrease in the frequency, duration and number of the oral ulcerations, it can also be monitored by a reduction of serum IL-6 level in RAU patients. PMID- 12653860 TI - Molecular epidemiology of invasive stomatitis due to Aspergillus flavus in patients with acute leukemia. AB - BACKGROUND: Invasive oral aspergillosis is a rare complication and only little information on the epidemiology of Aspergillus flavus infection is available. We present here the molecular analysis of the epidemiology of invasive stomatitis due to Aspergillus flavus in patients with acute leukemia. METHODS: During a 5 year period (1992-1996), six isolates of A. flavus were obtained from leukemic patients with invasive Aspergillus stomatitis. Random amplification of polymorphic DNA (RAPD) with three different PCR primers was carried out to investigate the DNA typing of the isolates. RESULTS: The molecular analysis using RAPD revealed that three isolates of A. flavus obtained in 1992 from three patients were of the same type, whereas each of the isolates from the other three patients had a distinct unique band, resulting in four groups of A. flavus. CONCLUSION: As the three patients with invasive oral aspergillosis detected in 1992 were infected by a single strain of A. flavus, the strain was suspected to have caused a nosocomial outbreak of invasive oral aspergillosis in the hematology unit. PMID- 12653861 TI - Helicobacter pylori DNA in recurrent aphthous stomatitis. AB - BACKGROUND: Considering not only the fact that recurrent aphthous stomatitis (RAS) and stomach ulcers are immunologically mediated ulcers associated with Helicobacter pylori, but also the recent evidence that anaemia can be associated with both diseases, and the discovery of H. pylori in the oral mucosa led us to hypothesize that this bacteria may be related to RAS pathogenesis. METHODS: Thirty-six consecutive subjects affected by minor and major forms of RAS and 48 healthy volunteers were included in the present study. The nested polymerase chain reaction (PCR) technique was used to detect the presence of H. pylori in the oral lesion, the normal contralateral mucosa of patients affected by RAS and the oral mucosa of control subjects. The chi2- and Fisher's tests were used for statistical analysis. RESULTS: No association between RAS lesions and H. pylori was observed. However, 14 out of 36 (38.9%) of the patients with RAS were found to show the presence of H. pylori DNA in the lesion and/or contralateral mucosa. Sixteen out of 48 (33.3%) of the patients without RAS (control subjects) were positive (P > 0.05). CONCLUSION: The present study does not give support to the assumption that H. pylori could be involved in RAS development. PMID- 12653863 TI - Evidence for the formation of metaplastic bone from pericytes in calcifying fibroblastic granuloma. AB - BACKGROUND: The calcifying fibroblastic granuloma (CFG) is a variant of epulis characterised histologically by calcified nodules and metaplastic bone formation and clinically by its pattern of frequent recurrence. Recent evidence suggests that calcified nodules and bone may form directly from non-haemopoetic pericytes in a variety of situations. This study aimed to test the hypothesis that metaplastic bone formation in CFG may be the result of a pericytic differentiation pathway. METHODS: A series of 16 cases of CFG meeting the criteria of Lee were reviewed and stained using markers which aid the identification of pericyte (smooth muscle actin, von Willebrand's factor, monoclonal antibody 3G5) and osteoblast (AML-3) lineages. RESULTS: The cells in mineralising areas of CFG resembled pericytes morphologically. Often, calcified nodules were observed within the vascular basement membrane and nodules frequently formed the nidus for reactive bone formation. The pericyte-like cells were positive for smooth muscle actin and AML-3 and were negative for von Willebrand's factor. CONCLUSIONS: These data provide circumstantial evidence for a pericytic lineage in CFG. PMID- 12653862 TI - Cellular mechanisms of osteoclast formation and lacunar resorption in giant cell granuloma of the jaw. AB - BACKGROUND: Giant cell granuloma (GCG) is an osteolytic tumour of the jaw which is characterised by the presence of both mononuclear and multinucleated (osteoclast-like) giant cell components. The nature of these component cells and the pathogenesis of the extensive osteolysis associated with this lesion is uncertain. METHODS: Using cell culture techniques and immunohistochemistry, we defined the phenotypic characteristics of the mononuclear and multinucleated cells present in four cases of GCG of the jaw. We also analysed the cellular and humoral factors associated with osteoclast formation and osteolysis in these tumours and determined whether GCG stromal cells are capable of supporting osteoclast formation. RESULTS: GCG-derived giant cells expressed the phenotypic characteristics of osteoclasts (TRAP+, VNR+, and calcitonin responsive) and were capable of lacunar resorption. In addition to macrophages, the mononuclear cell population contained numerous spindle-shaped stromal cells which proliferated in culture and expressed RANKL; these GCG-stromal cells were capable of supporting human osteoclast formation from circulating monocyte precursors. CONCLUSION: Our findings indicate that the giant cells in GCG of the jaw are osteoclast-like and formed from monocyte/macrophage precursors which differentiate into osteoclasts under the influence of RANKL-expressing mononuclear stromal cells found in this lesion. PMID- 12653864 TI - Immunohistochemical study of matrilin-1 in arthritic articular cartilage of the mandibular condyle. AB - BACKGROUND: The objective of this study was to investigate the expression of matrilin-1 in arthritic articular cartilage of the mandibular condyle by means of immunohistochemical methods. METHODS: Condylar cartilage specimens were obtained from temporomandibular joints (TMJs) of 12 patients with arthritis (osteoarthritis and internal derangement) (mean age 51.8 years; age range 28-71 years) and four patients with TMJ ankylosis (mean age 44.0 years; age range 16-64 years), diagnosed clinically and with imaging examinations. Paraffin sections were immunostained with anti-matrilin-1 antibodies. RESULTS: Matrilin-1 expression was detected in both patient groups with TMJ ankylosis and arthritis, and the level was remarkably higher in arthritic cartilage. The mean percentage of matrilin-1-producing cells to the total chondrocytes was significantly (P < 0.05) greater in the arthritic group (43.9 +/- 19.2%) than in subjects with TMJ ankylosis (28.0 +/- 8.7%). CONCLUSIONS: Articular chondrocytes in the TMJ condyle can express matrilin-1 and the expression is enhanced in arthritic cartilage, suggesting a presence of functional or adaptive remodeling in the condyle in response to degenerative changes in the TMJ structures. PMID- 12653865 TI - Detection of HHV-8 sequences and antigens in a MALT lymphoma associated with Sjogren's syndrome. AB - We describe the case of a bilateral parotid mucosa-associated lymphoid tissue (MALT) lymphoma associated with 2 years history of Sjogren's syndrome (SS), which was linked to human herpes virus 8 (HHV-8) infection. Using polymerase chain reaction (PCR) assay HHV-8 sequences were detectable in the lymphoma tissue of both sides. Serologic testing of the patient revealed HHV-8 antibodies in enzyme linked immunosorbent assay (ELISA) and immunofluorescence assay (IFA). Immunohistologic staining with two antibodies against open reading frame (ORF) 26 and v-cyclin homologues of HHV-8 revealed positive staining of the salivary acinic cells whereas the lymphoma cells were negative. The potential influence of HHV-8 infection for MALT lymphoma development in this case and possible parallels to gastric MALT lymphoma are discussed. PMID- 12653866 TI - Histopathological characteristics of eruption mesenchymal calcified hamartoma: two case reports. AB - Odontogenic calcified masses were present in the opercula of lower first molars that were delayed in eruption. The masses were relatively small, opaque, white in color with a smooth texture. Histopathological examinations revealed that they contained osteodentin, cementum, and pulp-like components; however, not odontogenic epithelial cells or enameloid. Further, mesenchymal multinucleated giant cells and dysplastic dental matrices were observed in the connective tissues surrounding the masses. These clinical and histopathological findings disagree with the features of pericoronal odontogenic hamartoma lesions, including odontoma, ameloblastic fibroma, and ameloblastic fibro-odontoma. Therefore, we propose to categorize this odontogenic mass as a new variety of hamartoma, eruption mesenchymal calcified hamartoma. PMID- 12653867 TI - Should pseudopolycythaemia be treated? PMID- 12653868 TI - DNA vaccines: a review. AB - The DNA vaccines are simple rings of DNA containing a gene encoding an antigen, and a promoter/terminator to make the gene express in mammalian cells. They are a promising new approach for generating all types of desired immunity: cytolytic T lymphocytes (CTL), T helper cells and antibodies, whilst being a technology that has the potential for global usage in terms of manufacturing ease, broad population administration and safety. This review gives an overview of the mechanisms, preclinical and clinical efficacy of DNA vaccines, and point out the limitations of the first generation of such vaccines, and some of the promising second-generation developments. This technology is also being utilized in the field of proteomics as a tool to elucidate the function of genes. The breadth of applications for DNA vaccines thus ranges from prophylactic vaccines to immunotherapy for infectious diseases, cancer, and autoimmune and allergic diseases. PMID- 12653869 TI - Pericardiocentesis guided by 2-D echocardiography: the method of choice for treatment of pericardial effusion. AB - BACKGROUND: Percutaneous pericardiocentesis guided by 2-D echocardiography has been used at Linkoping Heart Centre since 1983. AIM: To evaluate our experience of this method including a follow-up and also to determine the aetiology of pericardial effusion. METHODS: A retrospective study including 120 of 252 consecutive patients punctured. RESULTS: The two most common aetiologies were cardiac surgery (77% valve surgery), followed by malignant disease. The postsurgical effusions became clinically important a median of 12 days after surgery (range 0-56 days). The median survival in the group with malignant disease was 89 days (30-day survival 87%, 1-year survival 10%). Indwelling catheter was used in 93% of the patients. There was no mortality but one patient needed a second pericardiocentesis after an accidental puncture of the right ventricle. Nine patients had rhythm aberrations. Recurring effusion that needed puncture was seen in 8%. CONCLUSION: Pericardiocentesis guided by 2-D echocardiography is a safe and efficient method to treat pericardial effusion and also valuable as palliative treatment for patients with malignant aetiology of the effusion. PMID- 12653870 TI - Survival and trends of occurrence of left ventricular hypertrophy, gender differences, 1967-92: The Reykjavik Study. AB - OBJECTIVE: We estimated the prevalence, incidence and risk factors of left ventricular hypertrophy (LVH) in a prospective cohort study of 26 489 participants. MATERIAL AND METHODS: The LVH was defined as Minnesota Code 310 on electrocardiogram (ECG). Everyone with this code at first visit was defined as a prevalence case and those who developed it between subsequent visits were incidence cases. The comparison cohort were all other participants in the Reykjavik Study stages I-V. RESULTS: A total of 297 men and 49 women were found to have LVH of 3.2% and 0.5%, respectively. The incidence was 25 per 1000 per year amongst men and six per 1000 per year amongst women. Prevalence in both genders increased with increasing age. Risk factors at the time of diagnosis were systolic blood pressure [odds ratio (OR) per mmHg 1.02; 95% confidence interval (CI): 1.01-1.03], age (OR per year 1.04; 95% CI: 1.02-1.05), silent myocardial infarction (MI) (OR 3.18; 95% CI: 1.39-7.27) and ST-T changes (OR 3.06; 95% CI: 2.14-4.38) amongst men and systolic blood pressure and age for women with similar odds ratio. Predictive factors for acquiring LVH were systolic blood pressure [incidence ratio (IR) 1.01; 95% CI: 1.01-1.02] and angina with ECG changes (IR 2.33; 95% CI: 1.08-5.02) amongst men and systolic blood pressure amongst women (IR 1.03; 95% CI: 1.01-1.04). The risk for coronary mortality was significantly increased amongst women with hypertrophy [hazard ratio (HR) 3.07; 95% CI: 1.5 6.31] and their total survival was poorer with increasing time from diagnosis of LVH (HR 2.17; 95% CI: 1.36-3.48). CONCLUSIONS: We conclude that the presence of LVH and its appearance is associated with age and increased blood pressure amongst both genders. Women with LVH have poorer survival than other women and they are at threefold risk of dying of ischaemic heart disease. PMID- 12653871 TI - Plasma oxidized LDL: a predictor for acute myocardial infarction? AB - OBJECTIVES: Oxidized LDL has been attributed a key role in the development of atherosclerosis. Previous studies have demonstrated increased plasma levels of oxidized LDL in patients with established coronary artery disease. The aim of the present study was to investigate if plasma oxidized LDL also predicts risk for development of coronary heart disease (CHD). DESIGN: We used a nested case control design to study the association between plasma levels of oxidized LDL and risk for development of acute myocardial infarction (AMI) and/or death by CHD. SUBJECTS: Oxidized LDL was analysed by ELISA in cases (n = 26), controls (n = 26) and controls with LDL cholesterol >5.0 mmol L-1 (n = 26). RESULTS: Oxidized LDL correlated with total plasma and LDL cholesterol in both cases (r = 0.72, P < 0.01, r = 0.69, P < 0.01, respectively) and controls (r = 0.71, P < 0.01, r = 0.77, P < 0.01, respectively). The oxidized LDL/plasma cholesterol ratio was higher amongst cases (13.5, range 10.7-19.8) than in controls (12.6, range 9.5 15.8, P < 0.05) and hypercholesterolaemic controls (12.2, range 8.0-16.0, P < 0.01). CONCLUSIONS: These findings identify high plasma oxidized LDL/total cholesterol ratio as a possible indicator of increased risk for AMI. PMID- 12653872 TI - Multiple risk factor intervention reduces cardiovascular risk in hypertensive patients with echolucent plaques in the carotid artery. AB - OBJECTIVE: In a previously published randomized 6-year study we observed that multiple risk factor intervention reduced cardiovascular risk in high-risk hypertensive men, and that this effect was confined to patients with carotid artery plaques. Hypothetically, the underlying mechanism might have been a stabilization of echolucent, instable, rupture-prone plaques. The aim of the present study was to examine plaque characteristics by B-mode ultrasound in the previous intervention study, and also to investigate the relationship between plaque characteristics at baseline and cardiovascular events during the 6-year follow-up in the two randomization groups. METHODS: High resolution B-mode ultrasound was used to characterize plaque echogenicity in four subgroups - dominantly echolucent, substantially echolucent, dominantly echogenic, and uniformly echogenic. RESULTS: In the usual care group 17 of 32 (53%) patients with echolucent plaques at baseline suffered from a combined end-point (any death or nonfatal myocardial infarction or nonfatal stroke) during follow-up compared with seven of 28 (25%) patients in the intervention group (P = 0.036). The corresponding numbers in patients with echogenic plaques were n = 4/13 (31%) and n = 4/17 (24%), respectively (NS). In the usual care group 11 of 33 (33%) patients with no plaques suffered from a combined end-point during follow-up compared with 11 of 30 (37%) in the intervention group. CONCLUSION: Our data indicate that the beneficial effect of the multiple risk intervention programme was confined to those patients with echolucent plaques. The data have to be confirmed with a large-scale trial. PMID- 12653873 TI - Congestive heart failure is associated with the rate of bone loss. AB - AIMS: To characterize relationships between mineral homeostasis, bone turnover, bone mass, and congestive heart failure (CHF), we evaluated 75 women with mild to moderate CHF. METHODS AND RESULTS: We examined the association in annual rate of change in spinal bone mineral density (BMD) with polymorphism of the vitamin D receptor (VDR) gene. Compared with the control group, the CHF group had reduced left ventricular ejection fraction (LVEF: 68.2 +/- 7.5% vs. 60.2 +/- 12.9%; P = 0.0249), human atrial natriuretic peptide (hANP) was elevated (hANP: 10.7 +/- 4.7 pmol L-1 vs. 25.8 +/- 24.2 pmol L-1; P = 0.001) and had lower peak VO2 (22.3 +/- 7.5 mL kg-1 min-1 vs. 15.8 +/- 7.4 mL kg-1 min-1; P = 0.0429). The CHF patients with the VDR FF genotype had a significantly high annual rate of decrease in BMD. In the CHF patients with the VDR FF genotype, urinary calcium excretion (FECa) was elevated (1.40 +/- 0.91% vs. 2.39 +/- 1.40%; P = 0.028), and serum bone-type alkaline phosphatase (B-ALP) was reduced (62.6 +/- 13.7 IU L-1 vs. 47.0 +/- 18.6 IU L-1; P = 0.0123). Also, FECa was correlated positively with furosemide dose (R = 0.881; P = 0.0087) and hANP concentrations (R = 0.635; P = 0.0147) and negatively with DeltaBMD (R = 0.72; P = 0.044) in the CHF patients with the VDR FF genotype. CONCLUSION: The CHF patients with the VDR FF genotype have higher rates of bone loss. These patients may need to increase their calcium intake and BMD may need to be followed more carefully over time. PMID- 12653874 TI - High GAD65 autoantibody levels in nondiabetic adults are associated with HLA but not with CTLA-4 or INS VNTR. AB - OBJECTIVES: To explore the relationship between genetic background and antibody levels in a nondiabetic population. We evaluated if high levels of autoantibodies against the 65 kDa isoform of glutamic acid decarboxylase (GAD65Ab), were associated with high-risk genes, i.e. HLA, CTLA-4 and INS VNTR genes. DESIGN AND SUBJECTS: Seventy-five (M/F 39/36) subjects exceeding the 95th percentile of GAD65 autoantibody index and 75 age and sex matched subjects below the 95th percentile, randomly selected amongst participants in the Vasterbotten Intervention Programme. METHODS: The GAD65 Ab were measured in a radioligand binding assay. HLA class II typing was performed by an oligoblot hybridization method. CTLA-4 repeat length was analysed and divided into short forms and long forms. Class I and class III alleles of INS VNTR were detected. Differences in distribution were tested by Pearson chi-square with Yates correction. Odds ratios (OR) were used to compare groups calculated with Cochran's and Mantel-Haenszel statistics. RESULTS: The DQB1*0201-DQA1*0501-DRB1*03 haplotype was increased in subjects with high GAD65Ab levels (P = 0.04). This increase seemed to be explained by a difference in haplotype frequencies amongst men (P = 0.01). Calculating OR showed a significant association between the DQB1*0201-DQA1*0501 DRB1*03 haplotype and elevated levels of GAD65Ab in all subjects (OR 2.2, 95% CI 1.02-4.9) as well as in men (OR 4.6, 95% CI 1.3-15.9). There was no association between high levels of GAD65Ab and either INS VNTR or CTLA-4 polymorphisms. CONCLUSION: Our study suggests that adult males with the DQB1*0201-DQA1*0501 DRB1*03 haplotype tend to develop high GAD65Ab titres. As none of these subjects have developed diabetes these data suggest that HLA may be important in GAD65Ab formation but that additional factors are required for the progression to overt type 1 diabetes. PMID- 12653875 TI - The hypothalamus-pituitary function after pituitary stereotactic radiosurgery: evaluation of growth hormone deficiency. AB - OBJECTIVES: Radiation therapy to the pituitary gland means a considerable risk of developing hypopituitarism. The aim of the study was to investigate the growth hormone releasing hormone (GHRH)-growth hormone (GH)-insulin-like growth factor-I (IGF-I) axis after treatment with stereotactic radiosurgery to the pituitary because of Cushing's disease. SETTING: Inpatient ward in university clinic. SUBJECTS: Eleven adult patients (eight women, three men), 20-65 years of age were studied 2.5-11.3 years after stereotactic radiosurgery (isocentre dose 50-100 Gy lesion-1) and compared with healthy controls. MAIN OUTCOME MEASURES: Spontaneous GH secretion was evaluated as 12-h night GH profiles. Stimulated GH responses were evaluated in seven of 11 patients using arginine-insulin and GHRH tests. Serum IGF-I levels were measured in fasting serum morning samples. RESULTS: All patients except one displayed blunted nocturnal GH profiles. After arginine insulin challenge, six of seven patients displayed low GH release. GH response was higher after GHRH injection compared with both the response to arginine insulin and to the maximum GH levels in the nocturnal profiles. Seven patients had an IGF-I standard deviation score (SDS) within the normal range for age. Serum IGF-I values were correlated to mean GH values in the 12-h night profile (r = 0.67, P < 0.05) and both these variables were negatively correlated to time elapse since last radiation treatment (r = -0.64, P < 0.05 and r = -0.78, P < 0.05, respectively). CONCLUSIONS: Our patients with Cushing's disease evaluated several years after stereotactic radiosurgery as the primary and only treatment, demonstrated severely blunted spontaneous GH secretion and GH response to arginine-insulin. A disturbed regulation at the hypothalamic level was suggested as mechanism for this. Noteworthy is that serum IGF-I values correlated to the mean values of the 12-h GH profile. PMID- 12653876 TI - Intermittent versus on-demand use of a very low calorie diet: a randomized 2-year clinical trial. AB - OBJECTIVES: To compare two different very low calorie diet (VLCD)-based weight maintenance strategies. DESIGN AND SETTING: A randomized 2-year clinical trial performed at the Department of Body Composition and Metabolism, Sahlgrenska University Hospital, Sweden. SUBJECTS: A total of 334 patients, body mass index (BMI) >30 kg m-2, aged 18-60 years. INTERVENTIONS: All the patients started with 16 VLCD weeks. Subjects in the intermittent group were then scheduled to use VLCD for 2 weeks every third month, whilst patients in the on-demand group were instructed to use VLCD whenever their body weight passed an individualized cut off level. Irrespective of the treatment group, all the subjects were recommended a hypocaloric diet during VLCD-free periods. MAIN OUTCOME MEASURES: Changes in body weight, body composition, anthropometric variables and cardiovascular risk factors. RESULTS: Completers in both groups maintained highly significant weight losses after 2 years: 7.0 +/- 11.0 kg (6.2 +/- 9.5%) in the intermittent group and 9.1 +/- 9.7 kg (7.7 +/- 8.1%) in the on-demand group (P < 0.001, ns between groups). Male completers in the on-demand group lost significantly more weight than men in the intermittent group, 14.5 +/- 11.0 kg vs. 4.0 +/- 10.5 kg, respectively (P < 0.01). Most cardiovascular risk factors improved during the first year, whilst anthropometric measures, insulin, HDL- and LDL-cholesterol were also significantly improved after 2 years of treatment. CONCLUSION: Clinically significant weight reductions were achieved after 2 years of VLCD based treatment. The structure of VLCD treatment during the maintenance phase did not affect weight loss in the total study population, whilst male subjects might benefit from the VLCD on-demand strategy. PMID- 12653877 TI - Health economics in the Hypertension Optimal Treatment (HOT) study: costs and cost-effectiveness of intensive blood pressure lowering and low-dose aspirin in patients with hypertension. AB - OBJECTIVES: To investigate the marginal cost-effectiveness of different targets for the reduction of blood pressure and the cost-effectiveness of adding acetylsalicylic acid (ASA) to the treatment of hypertension. DESIGN: Patients with hypertension were randomized to three target groups for blood pressure; < or =90, < or =85 and < or =80 mmHg. Patients were also randomly assigned ASA and placebo. The average follow-up time was 3.8 years. The direct costs for drugs, visits, hospitalizations, and side-effects were calculated and related to clinical outcome. SETTING: Resource utilization data from all the 26 countries in the study were pooled, and Swedish unit costs were applied to the aggregated resource utilization. SUBJECTS: A total of 18 790 patients, 50-80 years of age (mean 61.5 years), with a diastolic blood pressure between 100 and 115 mmHg (mean 105 mmHg). INTERVENTIONS: Antihypertensive treatment with the long-acting calcium antagonist felodipine was given to all patients. Additional therapy and dose increments in four further steps were prescribed to reach the randomized target blood pressure. Fifty per cent of the patients were randomized to a low dose, 75 mg daily, of acetylsalicylic acid. MAIN OUTCOME MEASURES: Direct health care costs, major cardiovascular (CV) events (myocardial infarction and stroke) and CV death. RESULTS: The average cost of drugs and visits increased with more intensive treatment. The increase in treatment costs was partly but not fully offset by a nonsignificant reduction in the cost of CV hospitalizations. For patients with diabetes there were no significant differences in total cost between the target groups. The cost of avoiding a major CV event was negative in the base case analysis, SEK -10 360 (CI: -78 195, 75 630), and SEK 18 450 (CI: 88 789, 192 980) in a sensitivity analysis. For patients on ASA, costs were slightly but significantly higher than for patients on placebo. The estimates of the cost of avoiding a major CV event varied between SEK 41 600 and SEK 477 400, with very wide confidence intervals. CONCLUSIONS: The treatment cost increases as the target for hypertension treatment is lowered. In patients with diabetes, intensive treatment to a lower target is cost-effective. Because of the nonsignificant difference in events, no conclusion can be made for all patients in the study. Furthermore, no conclusive evidence was found regarding the cost effectiveness of adding ASA to the treatment of hypertension. PMID- 12653878 TI - Life-threatening thrombotic complications of relative polycythaemia. AB - Relative polycythaemia refers to raised haematocrit with normal red cell mass. Plasma volume may be reduced. This condition is associated with acute hypoxia, smoking, alcohol and diuretics. We describe two life-threatening thrombotic events in two patients with relative polycythaemia under age 40 years. The first had myocardial infarction and on admission haemoglobin was 21.6 g dL-1. The second developed pulmonary embolism and haemoglobin was 19.1 g dL-1. Both patients received antithrombotic measures and isovolumetric venesection. Sixteen patients (age <40 years) who attended our accident and emergency department in 1 year had a haemoglobin level of >18.0 g dL-1. Recognition of relative polycythaemia in at risk-individuals may help reduce thrombotic risk. PMID- 12653880 TI - Septal alcohol ablation and Heyde's syndrome revisited. AB - We describe a 57-year-old male with hypertrophic obstructive cardiomyopathy and occult blood loss. Septal alcohol ablation reduced the left ventricular outflow tract obstruction and eliminated recurrent bleeding. The case resembles Heyde's syndrome, i.e. the association between occult gastrointestinal bleeding and aortic valve stenosis, where resolution of blood loss after corrective aortic valve surgery has also been reported. PMID- 12653879 TI - Insulinoma presenting with hyperandrogenism: a case report and a literature review. AB - An 18-year-old woman presented with a 6-month history of amenorrhoea and hyperandrogenism. Three months later she developed several episodes of fasting hypoglycaemia and was subsequently diagnosed with an insulinoma. Hyperinsulinaemia was observed in association with an elevated serum testosterone level. Surgical removal of the insulinoma resulted in resolution of the clinical and biochemical features of the polycystic ovarian syndrome (PCOS). Polycystic ovarian syndrome is unusual in a patient having an insulinoma. The rarity of this association may be the result of the late age of onset of this type of tumour, intermittent secretion of excessive insulin by the tumour, the degree of hyperinsulinism or other factors extrinsic to the insulin receptor that may facilitate insulin activity. However, we could not discover how our patient differs in having had PCOS from the majority of women with insulinoma who do not. If other patients with insulinoma are subsequently found to have hyperandrogenism, then this tumour might be added to the differential diagnosis of causes of anovulatory cycles and hyperandrogenaemia, although rare the association would be uncommon. PMID- 12653881 TI - Is there a common cause of adenoma, focal nodular hyperplasia, and hemangioma of the liver? PMID- 12653882 TI - Can a non-invasive strategy for managing young dyspeptics be safely implemented in Asia? AB - Dyspepsia is a common problem in the Asia-Pacific region, with a prevalence rate ranging from 10-20%. It constitutes 2-5% of consultations with primary-care physicians and forms a major part of the gastroenterologists' workload. Although upper gastrointestinal endoscopy (UGIE) is the investigation of choice, no serious disease is present in the majority of patients and various other ways have been suggested, mainly in the West, to reduce the demand on the finite resources of UGIE services. The alternative methods to UGIE have been based on non-invasive detection of Helicobacter pylori in patients with dyspepsia, as the organism has been shown to be associated with most peptic ulcers and even gastric cancer. A positive H. pylori test in a patient with dyspepsia may not necessarily indicate serious disease, but H. pylori eradication eliminates the propensity for developing peptic ulcers and perhaps even cancer (not proven). In high-risk populations, non-invasive screening for H. pylori can even be considered a 'cancer test', as it can help target investigations in a selected group of patients. PMID- 12653883 TI - Immune function in patients with acute pancreatitis. AB - BACKGROUND: The aim of this study was to clarify the relationship between the balance of T-helper (Th)1 and Th2 cytokines, and the numbers of CD4+ T and CD8+ T cells, and was investigated, together with the plasma concentration of the antigen, an apoptosis marker, in patients with mild and acute pancreatitis (AP). METHODS: Plasma concentrations of soluble (s) CD4, sCD8, sIL-2-R, IL-12, IFN gamma and sFas antigen were measured by ELISA, and CD4+ T, and CD8+ T lymphocyte counts were measured by flow cytometry. RESULTS: Both CD4+ T and CD8+ T-cells were reduced in number; in the severe cases the reduction in the former was more pronounced. A significant positive correlation was noted among the concentrations of sCD4, sIL-2-R and IL-12, and a significant positive correlation was also found between sCD4 and sFas. During the early stage of AP, the concentrations of sCD4, sCD8, sIL-2-R, IL-12 and IFN-gamma increased more in the severe cases compared with those who had milder symptoms; however, these increases were moderated during the clinical course. CONCLUSION: We considered that these Th1 type CD4+ T cells probably induce the activation of macrophages and further pro-inflammatory reactions during the early stage of AP, as well as exerting direct cytotoxicity effects through Fas/Fas ligand expression. PMID- 12653884 TI - How reversible is hepatic functional impairment in autoimmune hepatitis? AB - BACKGROUND AND AIMS: We quantifed the short-term effects of immunosuppressive therapy on hepatic metabolic function in autoimmune hepatitis to establish how long it takes to achieve maximum functional improvement. METHODS: We studied 14 newly diagnosed patients with autoimmune hepatitis (12 type 1, two type 2) by antipyrine clearance and conventional liver tests, then repeated studies at 3-6 month intervals during the first 18 months of immunosuppressive therapy. RESULTS: Low values for antipyrine clearance were found in 13 of 14 cases; serum albumin concentration was low in four, bilirubin raised in eight and prothrombin time prolonged in four. Following immunosuppressive treatment for 3 months, antipyrine clearance improved by 98% (standard error of the mean 24%), which was proportionally greater than for serum albumin, bilirubin or prothrombin time. Antipyrine clearance and serum albumin continued to improve after 6-12 months of immunosuppressive treatment in several cases, whereas there were no further improvements in alanine aminotransferase (ALT), bilirubin and prothrombin time. CONCLUSIONS: In the short term, immunosuppressive therapy for autoimmune hepatitis markedly improves hepatic metabolic function, which is particularly striking for the sensitive metabolic test antipyrine clearance, but may also be seen with serum albumin. However, it may take up to 12 months to achieve maximal functional recovery. Management guidelines on autoimmune hepatitis should be extended to emphasize that changes in hepatic metabolic function, as well as ALT and gamma-globulin levels, be taken into consideration in the definition of remission. PMID- 12653885 TI - Regulation of p21 and p27 expression by the hepatitis B virus X protein and the alternate initiation site X proteins, AUG2 and AUG3. AB - BACKGROUND: The hepatitis B virus X gene has three in-frame start codons encoding the pX, AUG2 and AUG3 proteins. The AUG2 and AUG3 genes are 5'-truncated in respect to the full-length pX gene; however, all three genes terminate at the same stop codon. The activity of pX as an oncogene is well characterized; however, less is known about the AUG2 and AUG3 proteins. METHODS: The effects of pX, AUG2 and AUG3 on p21Cip,1/WAF,1/MDA6 and p27Kip-1 cyclin kinase inhibitor (CKI) protein expression, and the impact they have on proliferation, were investigated in CHO K-1 cells. CHO K-1 cells were chosen because they can be transfected at 100% efficiency. RESULTS: p21- and p27-luciferase reporter expression is modulated by increasing doses of the hepatitis B X proteins. At low concentrations of pX or AUG2, p21- and p27-luciferase activity was increased, and at high concentrations, p21- and p27-luciferase activity was decreased. Expression of the AUG3 gene showed a different profile: it was increasingly stimulatory with dose for both promoters. Western blot analyses demonstrated that p21 and p27 protein levels were modulated as predicted based on data generated in the promoter-luciferase experiments. Tritiated thymidine labeling of DNA showed biphasic kinetics of incorporation in the presence of varying pX and AUG2 concentrations, whereas labeling decreased with AUG3 concentration. The growth inhibitory effect of pX expression was reduced by antisense ablation of either p21 or p27. CONCLUSIONS: The relative expression level of pX, AUG2, and AUG3 impacts on CKI expression and cell proliferation. Our findings may explain why divergent effects of pX expression on growth have been observed by different groups, which may be related to relative pX expression levels. PMID- 12653886 TI - Steatosis in chronic hepatitis C: association with increased messenger RNA expression of collagen I, tumor necrosis factor-alpha and cytochrome P450 2E1. AB - BACKGROUND: Increased levels of tumor necrosis factor (TNF)-alpha and oxidative stress have been implicated as factors contributing to hepatic injury in fatty liver diseases. As steatosis is associated with an accelerated progression of fibrosis in chronic hepatitis C (HCV), we hypothesized that the messenger (m)RNA expression of genes involved with the production of reactive oxygen species, inflammation and cellular injury would be increased in liver tissue from subjects with steatosis and chronic HCV. METHODS: Real-time polymerase chain reaction was performed to determine relative mRNA expression levels of collagen I, TNF-alpha, cytochrome P450 2E1 (CYP 2E1), transforming growth factor-beta1 and CD14 in liver biopsies from 38 patients with chronic HCV. The mRNA expression levels were compared between subjects with and without steatosis, fibrosis, and inflammation. RESULTS: Multivariate analysis demonstrated that collagen I mRNA expression was increased by 199% in steatosis (P = 0.02), 85% in moderate to severe fibrosis (P = 0.02) and 157% in inflammation (P = 0.03). Livers of patients with steatosis also had an increase in TNF-alpha mRNA expression by 50% (P = 0.03) and CYP 2E1 expression by 37% (P = 0.04) compared with non-steatotic livers. Tumor necrosis factor-alpha protein was localized to Kupffer cells, bile ducts and portal inflammatory cells by immunohistochemistry. CONCLUSION: Increased expression of TNF-alpha may be involved in the pathogenesis of liver injury and progression of fibrosis in individuals who have steatosis in association with chronic HCV. PMID- 12653887 TI - Transforming growth factor beta in hepatitis C virus infection: in vivo and in vitro findings. AB - BACKGROUND: Hepatitis C virus (HCV) is a leading cause of chronic liver disease (CLD) worldwide. The chronicity is a result of viral persistence and the ability of the virus to escape from the immune mechanisms of the host. Transforming growth factor (TGF)-beta is a cytokine thought to be responsible for viral persistence and liver fibrogenesis. METHODS: The present study examined the levels of TGF-beta messenger (m)RNA by reverse transcription polymerase chain reaction (RT-PCR) in 35 liver biopsies and HCV-transfected HepG2 cells. RESULTS: Transforming growth factor-beta mRNA was detected in nine liver biopsies from patients with chronic HCV infection, but was not detected in patients with non HCV-related CLD or controls. On quantitation by semiquantitative PCR, TGF-beta mRNA levels ranged from 10-4.75 to 10-12.8 amol (10-7.46 +/- 3.771) in liver biopsies of HCV-related CLD. No significant difference in TGF-beta receptor levels was observed by RT-PCR in HCV- or non-HCV-related CLD by immunohistochemistry. To correlate these findings with in vitro experiments, levels of TGF-beta mRNA and its receptors were determined by RT-PCR in HepG2 cells transfected with HCV and hepatitis B virus (HBV) constructs, using mock transfected cells as control. The TGF-beta protein levels were quantitated in these cell supernatants by enzyme immunoassay. The TGF-beta mRNA and protein levels were two logs and approximately 30 times higher in HCV-transfected HepG2 cells than in HBV- and mock-transfected cells, respectively. The TGF-beta receptors in HepG2 cells were also downregulated in HCV-transfected cells as compared with mock-transfected cells. CONCLUSION: These observations suggest upregulation of TGF-beta in HCV infection and a probable role for TGF-beta in the pathogenesis of HCV-related CLD. PMID- 12653888 TI - Hepatitis C virus genotypes and hepatic fibrosis regulate 24-h decline of serum hepatitis C virus RNA during interferon therapy in patients with chronic hepatitis C. AB - BACKGROUND AND AIMS: Recently, hepatitis C virus (HCV) dynamics during interferon (IFN) therapy have been studied in detail. We examined factors that regulate the viral kinetics and the relationship between the viral kinetics and clinical effect of IFN therapy. METHODS: Eighty-eight patients with chronic hepatitis C entered this study. All patients had been treated with 3 MU of IFN-beta twice a day for the first 2-4 weeks, then IFN-alpha for the next 20-22 weeks (three injections per week). The levels of serum HCV RNA were determined by Amplicor HCV Monitor version 1.0, before and 24 h after the first injection of IFN; then the decline of HCV was calculated. Liver inflammation and fibrosis were scored as 0 (none), 1 (mild), 2 (moderate) or 3 (severe) using biopsy specimens. RESULTS: The decline of serum HCV RNA was 1.42 +/- 0.65 log copies/mL in genotype 1b and 1.83 +/- 0.72 in genotype 2a or 2b (P < 0.01). By a logistic regression model, genotype (1b, 2a or 2b) and hepatic fibrosis (0 or 1, 2 or 3) associated with 24 h decline of serum HCV RNA, independently. As the predictor of IFN therapy, the decline of serum HCV RNA and serum HCV RNA levels before IFN therapy were the independent significant factors (P < 0.001). CONCLUSIONS: The decline of serum HCV RNA during the first 24 h of IFN therapy was regulated by genotypes and hepatic fibrosis. The decline of serum HCV RNA and initial HCV load were independent factors that can be the predictor of the subsequent sustained viral response to IFN therapy. PMID- 12653889 TI - Efficacy of interferon-alpha treatment in Japanese children with chronic hepatitis C. AB - BACKGROUND: We investigated the efficacy of natural interferon (IFN)-alpha treatment in 34 Japanese children with chronic hepatitis C. METHODS: Thirty-four children completed 6 months of therapy with natural IFN-alpha and were followed for 12 months or longer. We examined the serum hepatitis C virus (HCV) RNA titer and liver histology before, during, and after IFN treatment. RESULTS: At 6 months after the cessation of IFN-alpha treatment, 16 patients (47%) had normal serum alanine aminotransferase concentration and no detectable serum HCV RNA. There were no major side-effects, excluding some influenza-like symptoms during the IFN alpha treatment. Most genotype 2a patients had a complete response (80%). Moreover, patients who had a low HCV RNA titer (<102 copies/mL) after 1 month of IFN-alpha treatment became complete responders at 6 months after the cessation of treatment. Histological improvement was observed in almost all patients after IFN alpha treatment. CONCLUSION: Interferon-alpha treatment is safe and effective for children with chronic hepatitis C and has no serious side-effects. A HCV RNA concentration of <102 copies/mL after 1 month of IFN-alpha treatment and genotype 2a may be useful predictors of long-term IFN efficacy. PMID- 12653890 TI - Sarcomatous hepatocellular carcinoma: a special reference to ordinary hepatocellular carcinoma. AB - BACKGROUND: Sarcomatous hepatocellular carcinoma (HCC) has a worse prognosis than ordinary HCC. The relationship between the malignant potential of sarcomatous HCC and cell proliferation or cell adhesion is unknown. This study was undertaken to clarify this relationship. METHODS: In 21 cases of sarcomatous HCC, including 16 surgically resected and five autopsy cases, immunohistochemistry was used to compare the sarcomatous component (s-comp) and the ordinary component (o-comp) within each sarcomatous HCC. RESULTS: We found 15 epithelial-cadherin (E cadherin)-positive cases in o-comp (79%) and nine positive cases in s-comp (43%). The difference between sarcomatous HCC and ordinary HCC in E-cadherin-positive tumor prevalence was significant (P < 0.05). The Ki-67 (MIB1) labeling index ratio was 127 +/- 40 in s-comp and 80 +/- 33 in o-comp, and there was a greater tendency to have an ability to multiply in s-comp than in o-comp (P = 0.096). CONCLUSION: This study indicated that the loss of E-cadherin related to a morphological alteration in sarcomatous HCC; however, no relationship in respect to the malignant potential was found. PMID- 12653892 TI - Loss of heterozygosity of chromosome 8p and 11p in the dysplastic nodule and hepatocellular carcinoma. AB - BACKGROUND AND AIM: In hepatocarcinogenesis, both de novo and multistep pathways have been suggested, and in the latter a dysplastic nodule is the proposed precancerous lesion. But genetic changes involved in the dysplastic nodule are not well understood. In this study, we tried to determine whether allelic loss of the chromosome 8p and/or 11p could be involved in the development of the dysplastic nodule and/or hepatocellular carcinoma. Platelet-derived growth factor receptor beta-like tumor suppressor gene (PRLTS) and deletion in liver cancer-1 tumor suppressor gene are located at 8p21.3-p22. The hepatitis B virus integration site and WT1 tumor suppressor gene are located at 11p13. METHODS: We therefore studied loss of heterozygosity (LOH) of chromosome 8p21.3-p22 and 11p13 in 22 dysplastic nodules and 21 hepatocellular carcinomas. The samples, microdissected from paraffin-embedded tissues, were examined using a polymerase chain reaction-based LOH assay using microsatellite markers. RESULTS: Loss of heterozygosity was detected for chromosome 8p21.3-p22 in nine (40.9%) of 22 dysplastic nodules and in eight (42.1%) of 19 hepatocellular carcinomas. D8S261, located adjacent to PRLTS, showed most frequent LOH: 28.6% in dysplastic nodule and 40.0% in hepatocellular carcinoma. Loss of heterozygosity on chromosome 11p13 was found in three (15.8%) of 19 dysplastic nodules and in six (31.6%) of 19 hepatocellular carcinomas. Loss of heterozygosity of D11S995 and D11S907 was found in 33.3% and 7.1% of dysplastic nodules, and 8.3% and 44.4% of hepatocellular carcinomas, respectively. CONCLUSION: These results suggest that at least one putative tumor suppressor gene involved in the development and progression of hepatocellular carcinoma may be located on 8p21.3-p22 and 11p13. Particularly, PRLTS might be related to an early genetic event of hepatocarcinogenesis. PMID- 12653891 TI - Comparison of Doppler ultrasonography and the hepatic venous pressure gradient in assessing portal hypertension in liver cirrhosis. AB - BACKGROUND AND AIM: This prospective study aimed to determine whether Doppler ultrasonography can represent the hepatic venous pressure gradient (HVPG) as an assessment of the severity of portal hypertension and the response to terlipressin, which reduces the portal pressure in liver cirrhosis. METHODS: The HVPG and the Doppler ultrasonographic parameters, such as the portal venous velocity and the splenic venous velocity, the pulsatility and the resistive index of the hepatic, splenic and renal arteries were measured in 138 patients with liver cirrhosis. The changes in the HVPG and the portal venous velocity after administering terlipressin were evaluated in 43 of the 138 patients. The patients who showed a reduction in the HVPG of more than 20% of the baseline were defined as responders to terlipressin. RESULTS: None of the Doppler ultrasonographic parameters correlated with the HVPG. Both the HVPG (28.0 +/- 19.8%) and the portal venous velocity (29.7 +/- 13.2%) showed a significant reduction after terlipressin administration. However, the portal venous velocity decreased significantly, not only in the responders (31.0 +/- 12.0%) but also in the non responders (25.2 +/- 16.4%). CONCLUSIONS: Doppler ultrasonography does not represent the HVPG, and is therefore not suitable for replacing HVPG as a means of assessing the severity of portal hypertension and the response to drugs which reduce the portal pressure in liver cirrhosis. PMID- 12653893 TI - In vivo inefficiency of pentoxifylline and interferon-alpha on hepatic fibrosis in biliary-obstructed rats: assessment by tissue collagen content and prolidase activity. AB - BACKGROUND AND AIM: To evaluate the possible antifibrotic effects of two drugs, pentoxifylline (PTX) and interferon (IFN)-alpha as well as their combination, on a bile-duct-ligated rat hepatic fibrosis model. METHODS: Bile ducts of 34 female Wistar rats were ligated, and 24 bile ducts were sham operated. Bile-duct-ligated rats were divided into four groups, in which either sterile saline, IFN-alpha (100 000 IU/3 days a week), PTX (50 mg/kg/day) or IFN-alpha + PTX were administered. Sham-operated rats were treated at the same doses. On the 28th day, rats were decapitated to obtain blood for the measurements of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma-glutamyl transpeptidase (GGT) and bilirubins. Serum prolidase was assayed at the beginning and at the end of the study by the modified Chinard's colorimetric method. Liver prolidase was assayed after tissue homogenization. Liver collagen content was determined by the dye elution method described by Lopez de Leon. Morphometric-densitometric measurements of hepatic fibrosis were quantified by computerized image analysis. RESULTS: The AST, ALT, ALP, GGT and bilirubins, liver prolidase enzyme activity, collagen content and hepatic collagen surface density were found to be increased in bile-duct-ligated rats on day 28. There was no statistically significant recovery or even a change in collagen turnover rate in rats treated with alternate regimens applied in the study (P > 0.05). CONCLUSION: Pentoxifylline, IFN-alpha and their combination have no beneficial effect on experimental fibrosis induced by biliary obstruction. PMID- 12653894 TI - Inducibility of microsomal liver function may differentiate cirrhotic patients with maintained compared with severely compromised liver reserve. AB - BACKGROUND AND AIM: Quantitative tests of liver function (QTLF) can be modulated by enzyme-inducing agents. The objective of the study was to examine changes in QTLF after treatment with Phenobarbital, a potent cytochrome P450-inducing agent. METHODS: Forty-six consecutive patients with liver cirrhosis Child-Pugh score B and C (34 alcoholic, 12 hepatitis C) and a compromised aminopyrine breath test (ABT) were included. Thirty-six patients (group I) received phenobarbitone (150 mg) for 7 days. Ten patients (group II) received a placebo. The QTLF, which included ABT (microsomal liver function), galactose elimination capacity (GEC, cytosolic liver function), sorbitol clearance (SCl, liver plasma flow) and indocyanine green clearance (ICG, liver perfusion) was carried out before and after pharmacological induction. RESULTS: Group I showed a basal ABT of 0.18 +/- 0.11% dose.kg/mmol CO2 (normal >0.6%), which increased significantly after induction (172%, P < 0.05), whereas in group II the ABT values did not change. In group IB, a subgroup of group I which exceeded the basal threshold value of 0.30% after stimulation (n = 22), the ABT values increased significantly to 0.44 +/- 0.17% (244%, P < 0.01). The GEC, SCl and ICG remained unchanged before and after induction. CONCLUSIONS: After pharmacological induction, microsomal liver function increased significantly in a subgroup of patients with liver cirrhosis, whereas the GEC, SCl and ICG remained unchanged. Inducibility of the microsomal liver function could be used as a novel parameter of functional hepatic reserve and prognosis in cirrhosis. PMID- 12653895 TI - Randomized controlled trial to compare the dose of adjuvant chemotherapy after curative resection of hepatocellular carcinoma. AB - BACKGROUND AND AIM: Adjuvant locoregional chemotherapy has been shown to be useful to prevent recurrence after curative resection of hepatocellular carcinoma (HCC) in some retrospective studies. Our aim was to compare the dose effect in the prevention of tumor recurrence. METHODS: A prospective randomized controlled trial was conducted in patients with curative resection of HCC; they were given either one intra-arterial dose of cisplatin/lipiodol, or received four doses, once every 3 months. The rates of recurrence, disease-free and overall survival were compared. RESULTS: During a median follow up of 818 days, 21 patients received one dose and 19 received four doses, with 10 (47.6%) and eight (42.1%) recurrences, respectively. The 1-year, 2-year and 3-year disease-free survival rates were 71%, 54% and 44% for the one-dose group and 74%, 60% and 40% for the four-dose group (P = 0.78). The respective overall survival rates were 85%, 74%, 55% and 84%, 71%, 40% (P = 0.64). The only prognostic factor was presence of vascular permeation. The side-effects were mild and tolerable. CONCLUSIONS: There is no significant difference in the survival rates between the two groups. Adjuvant chemotherapy may not be useful. PMID- 12653896 TI - Gastrointestinal: synchronous and metachronous colorectal cancers. PMID- 12653897 TI - Hepatobiliary and pancreatic: right-upper-quadrant pain. PMID- 12653898 TI - Chronic hepatitis C associated with monoclonal gammopathy of undetermined significance. AB - BACKGROUND: Two patients were admitted to Mie Prefectural General Medical Center and diagnosed as chronic hepatitis C complicated with monoclonal gammopathy of undetermined significance (MGUS). METHODS: The MGUS class were immunoglobulin (Ig)G. The hepatitis C virus (HCV) RNA genotype was Ib. Based on these findings, they were diagnosed as chronic hepatitis C complicated with MGUS. RESULTS: Histological studies showed chronic hepatitis in the liver and a mild rise in plasma cells without dysplasia and abnormalities in the bone marrow. Serum examination for cryoglobulin was negative. CONCLUSION: Chronic HCV infection might play a pathogenic role in the multistage process leading to lymphoproliferative disorders. PMID- 12653900 TI - Idiopathic thrombocytopenic purpura with acute hepatitis C viral infection. PMID- 12653901 TI - Gallbladder aspiration for acute cholecystitis in high-surgical-risk patients. PMID- 12653902 TI - Pregnancy and exposure to infliximab (anti-tumor necrosis factor-alpha monoclonal antibody). PMID- 12653903 TI - Esophageal injury as a result of ingestion of iron tablets. PMID- 12653904 TI - Elevated carbohydrate antigen 19-9 levels in a patient with choledocholithiasis. PMID- 12653905 TI - Primary bile duct carcinoma in histocompatibility antigen-identical twins. PMID- 12653909 TI - Henoch-Schonlein purpura nephritis. PMID- 12653910 TI - Carotenoderma--a review of the current literature. PMID- 12653911 TI - Chancroid: contemporary appraisal. PMID- 12653912 TI - Correlating skin phototype and minimum erythema dose in Arab skin. AB - Skin typing, as proposed by Fitzpatrick, has been used for estimating the initial dose in phototherapy.1 However, several reports have questioned the reliability of the method.2-4 It has been suggested that skin typing should not be the sole basis for starting UV-B phototherapy, and that determining the minimum erythema dose (MED) is a more reliable method of estimating the UV-B dosage.5 There is little published data with respect to skin types and MED values in Arab skin. In this review, we present our experience on the subject with Bahraini patients. PMID- 12653913 TI - A seroepidemiologic survey of the prevalence of varicella-zoster virus in the pediatric population in two university hospitals in Brazil. AB - BACKGROUND: Varicella-zoster virus (VZV) is an alpha-herpesvirus causing varicella (chickenpox) and herpes zoster (shingles). Varicella results from primary VZV infection, and is a common childhood illness associated with fever and a generalized, pruritic, vesicular eruption. Herpes zoster is caused by VZV reactivation later in life (most cases after the fifth decade), and is characterized by a localized, painful, and vesicular eruption involving one or adjacent dermatomes. The incidence of herpes zoster increases with age and immunosuppression. OBJECTIVE: To estimate the seroprevalence of VZV in the Brazilian pediatric population by evaluating the prevalence of specific antibodies to VZV in children from two university hospitals in the state of Rio de Janeiro. METHODS: A population composed of 160 children derived from two university hospitals in the state of Rio de Janeiro was included in the study. All patients completed a questionnaire regarding their socio-epidemiologic characteristics, and a complete physical examination was performed. All blood samples were screened using a commercial enzyme-linked fluorescent assay (ELFA) kit, specific for the detection of immunoglobulin G (IgG) antibodies to VZV. RESULTS: The seroprevalence of VZV was 58.1% in the overall population, with a statistically significant correlation between seroprevalence and increasing age (P < 0.0001). A previous history of measles infection (P < 0.04), previous history of varicella infection (P < 0.0001), and the presence of skin lesions (P < 0.05) were significantly associated with seropositivity to VZV. CONCLUSIONS: Further studies should be performed in order to evaluate the endemicity of VZV infections and to establish criteria for the use of the specific vaccine in Brazil. PMID- 12653914 TI - Incidence of cutaneous tuberculosis in patients with organ tuberculosis. AB - BACKGROUND: Tuberculosis continues to be a health problem in some countries. The development of resistance to antituberculitic drugs and the increase in diseases and conditions associated with immunodeficiency such as AIDS and chemotherapy have caused tuberculosis to increase recently. As a result, the incidence of cutaneous tuberculosis has been increasing as well. AIM: To detect cutaneous tuberculosis in patients with organ tuberculosis and to establish some characteristics of the relation between organ and cutaneous TB. MATERIAL AND METHODS: A total of 370 patients (145 females and 225 males), aged 2-76 years (mean age 27.5), enrolled for this screening study. These patients were hospitalized patients who already had pulmonary or extrapulmonary tuberculosis diagnosed before admission. All patients underwent a general skin examination, and, if needed, cutaneous biopsies were taken from involved skin areas. RESULTS: Three hundred and forty-seven (93.78%) out of 370 patients had pulmonary tuberculosis only or in association with one of other organ tuberculoses. Twenty three patients had extrapulmonary TB: nine were TB adenitis, six were TB peritonitis, three were bone tuberculosis, and five were TB meningitides. Of 370 patients, only 13 (3.51%) had cutaneous TB: seven scrofuloderma (SCD; 2.16%), four lupus vulgaris (LV; 1.35%), one LV and SCD, and one Bacille Calmette-Guerin (BCG) adenitis (0.027%). Cutaneous tuberculosis was observed in seven out of 260 patients with parenchymal tuberculosis (2.96%). Four out of nine patients with TB adenitis (44.4%), one out of 12 pulmopleuretic (8.3%), and one out of 67 pleuresic patients (1.40%) had cutaneous TB as well. Mean age of the 13 patients was 32.46 years: mean age of SCD and LV was 24.8 and 48 years, respectively. The one patient with BCG adenitis was 7 months old. Five (62.5%) out of eight patients with SCD, and only one (20%) out of five patients with LV were new cases. Four patients with SCD had a positive family history, while LV patients did not. CONCLUSIONS: Organ tuberculosis is rarely associated with cutaneous tuberculosis. Scrofuloderma and LV are the most frequent forms of skin TB associated with organ TB in this population. Tuberculosis adenitis is the organ TB that causes cutaneous TB most often among other organ tuberculoses. More than one form of cutaneous TB affected only one patient with pulmonary TB; therefore, it is very rare. Tuberculids were not observed in any of the patients. PMID- 12653915 TI - Eyelid trichoblastoma: an unusual localization. PMID- 12653916 TI - American cutaneous leishmaniasis unresponsive to antimonial drugs: successful treatment using combination of N-methilglucamine antimoniate plus pentoxifylline. AB - BACKGROUND: American cutaneous leishmaniasis is characterized by single or multiple ulcerations. Cytokines, among other factors, have been shown to influence lesion development and tumoral necrosis factor-alpha is a major cytokine implicated in pathogenesis of ulcers. OBSERVATIONS: We tested oral pentoxifylline, a known tumoral necrosis factor-alpha inhibitor, at doses of 400 mg (2-3x/day), associated to N-methylglucamine antimoniate (15 mg/kg/day) in two patients with American cutaneous leishmaniasis unresponsive to antimonial drugs. We observed a satisfactory response with quick cure of skin lesions of these patients. CONCLUSIONS: Our results suggest that oral pentoxifylline in association to N-methylglucamine antimoniate should be consider in refractory cutaneous leishmaniasis patients. PMID- 12653917 TI - Disseminated gonococcemia. AB - A 26-year-old woman presented with a high-grade fever and chills of 2 days' duration. She complained of associated joint pain, especially in the wrists and knees. One day before admission, tender skin lesions began to develop on the fingers, and subsequently spread to the more proximal extremities. The patient recalled having a sore throat and a nonproductive cough before the onset of the fever and eruption. The past medical history was significant for Gardnerella vaginitis and several urinary tract infections. The patient was taking oral contraceptive pills; her most recent menstruation was 3 weeks before admission. She reported having sexual intercourse with her boyfriend 2 weeks before admission. The patient's temperature was 40 degrees C. Dermatologic examination revealed a 6-mm, hemorrhagic pustule on an ill-defined pink base, overlying the volar aspect of the left second proximal interphalangeal joint (Fig. 1a). Scattered on the upper and lower extremities were occasional round, ill-defined pink macules with central pinpoint vesiculation (Fig. 1b). A skin biopsy of the digit revealed a dense neutrophilic infiltrate with leukocytoclasis and marked fibrin deposition in the superficial and deep dermal vessels (Fig. 2a). Gram stains demonstrated the presence of Gram-negative diplococci (Fig. 2b). Laboratory findings included leukocytosis (leukocyte count of 20 x 109/L, with 81% neutrophils). Analysis of an endocervical specimen by polymerase chain reaction was positive for Neisseria gonorrhoeae and negative for Chlamydia trachomatis. Throat and blood cultures grew N. gonorrhoeae. Specimen cultures obtained by skin biopsy yielded no growth. Results of serologic analysis for human immunodeficiency virus, hepatitis, syphilis, and pregnancy were negative. Beginning on admission, intravenous ceftriaxone, 2 g, was administered every 24 h for 6 days, followed by oral cefixime, 400 mg twice daily for 4 days. Oral azithromycin, 1 g, was administered to treat possible coinfection with C. trachomatis. By treatment day 4, the patient was afebrile, with the resolution of leukocytosis and symptomatic improvement of arthralgias. PMID- 12653918 TI - Barraquer-Simons syndrome with benign infundibulocystic proliferation. PMID- 12653919 TI - Pyoderma gangrenosum, acne conglobata, and IgA gammopathy. PMID- 12653920 TI - Erosive perianal lichen planus responsive to tacrolimus. PMID- 12653921 TI - Reconstruction of epidermis by grafting of keratinocytes cultured on polymer support--clinical study. AB - BACKGROUND: Extensive wound coverage still represents a challenge for contemporary medicine. We demonstrate the results of a clinical trial of the grafting of cultured keratinocytes directly on a polymer cultivation support in the treatment of skin defects in seriously burned patients and in patients with trophic ulcers. METHODS: Wound closure was evaluated clinically. The morphology and phenotypic pattern of the reconstructed epidermis, including the basal lamina, as well as the presence of Langerhans cells, were evaluated immunocytochemically using a panel of monoclonal antibodies. RESULTS: All layers of the reconstructed epidermis were normally differentiated (cytokeratin immunocytochemistry). The basal lamina contained collagen type IV and laminin. The reconstructed epidermis was extensively colonized by Langerhans cells. CONCLUSIONS: The results of the described technology are encouraging, especially in patients after a burn injury. The described procedure is suitable for the treatment of skin defects in clinical practice. PMID- 12653922 TI - Repeat courses of intravenous alefacept in patients with chronic plaque psoriasis provide consistent safety and efficacy. AB - BACKGROUND: Psoriasis is a chronic, relapsing skin disease that may require multiple treatment courses. Alefacept targets the memory T cells implicated in psoriasis pathogenesis. This open-label study evaluated the safety and tolerability, efficacy, and pharmacodynamics of repeat courses of alefacept in men and women with chronic plaque psoriasis. This article reports the interim results of this ongoing study. METHODS: Patients (n = 174) who participated in previous phase II studies of alefacept were included in this retreatment study. Intravenous alefacept (7.5 mg) was administered once weekly for 12 weeks followed by 12 weeks of observation. Initial and subsequent retreatment courses were only given when, in the opinion of the investigators, disease had returned and necessitated treatment; CD4+ T-cell counts had to be at or above the lower limit of normal. RESULTS: Adverse events were similar regardless of the retreatment course. No opportunistic infections, rebound of disease, or flares were reported. Low titers of anti-alefacept antibodies occurred in a few patients without related safety issues. Sixty-six per cent of patients achieved a >/= 50% reduction in the Psoriasis Area and Severity Index (PASI) at any time after the first dose of retreatment course 1. Patients who received two retreatment courses (n = 50) had consistent or improved responses after the second course; 64% and 68% of these patients achieved a >/= 50% PASI improvement at any time after the first dose of retreatment courses 1 and 2, respectively. Alefacept selectively reduced memory T cells without cumulative effects. CONCLUSIONS: Repeat courses of alefacept were well tolerated, and subsequent retreatment courses were at least as effective as the initial course of therapy. PMID- 12653923 TI - Newly available treatments for psoriatic arthritis and their impact on skin psoriasis. AB - Far from being a "benign" arthropathy, as it was initially characterized, psoriatic arthritis (PsA) affects approximately 0.2% of the US population and can be associated with considerable joint damage, symptomatology, and quality of life impairment. PsA shares many characteristics with rheumatoid arthritis (RA), and new, rationally designed drugs that are effective in RA also are proving active in PsA. Two such drugs, etanercept and infliximab, target tumor necrosis factor (TNF), a key component of the inflammatory response. This review discusses the rationale for and experience with the use of these agents in PsA. Etanercept is a dimeric fusion protein that binds specifically to TNF, blocking its interaction with cell surface TNF receptors. Infliximab is a chimeric (murine/human) monoclonal antibody that binds to TNF and inhibits its binding to its receptor. A randomized placebo-controlled trial of etanercept in PsA found statistically significant benefits for this agent in measures of arthritic activity and psoriatic severity. There have been anecdotal reports of the efficacy of infliximab in PsA, but results from controlled clinical trials of this agent in PsA have not been reported. TNF inhibitors represent new therapeutic options for patients with PsA. The potential advantages of treatment with etanercept and infliximab early in the disease course are discussed. PMID- 12653924 TI - Long-term control of mycosis fungoides of the hands with topical bexarotene. AB - BACKGROUND: Limited Stage IA mycosis fungoides (MF) is often treated with topical steroids, which can cause atrophy, or with nitrogen mustard, which imposes several limitations on the patient's lifestyle. Topical bexarotene is a novel synthetic rexinoid with few side-effects that has shown efficacy for treatment of mycosis fungoides skin lesions in recent Phase II-III clinical trials. The Phase I-II trial involving 67 stage IA-IIA MF patients demonstrated complete response (CR) in 21% and partial response (PR) in 42% of the patients. The median time to response was approximately 20 weeks. In the phase III trial of refractory stage IA, IB and IIA MF, the patients demonstrated a 44% response rate (8% CR). Patients with no prior therapy for mycosis fungoides responded at a higher rate (75%) than those with prior topical therapies. METHODS: Case report of a patient with MF limited to the hands treated with topical bexarotene 0.1% gel in a open label phase II clinical trial. RESULTS: Partial response occurred after 2 weeks of topical bexarotene therapy and the lesions were well controlled for 5 years using bexarotene monotherapy, with only occasional mild local irritation. CONCLUSIONS: Topical bexarotene is effective as long-term treatment monotherapy for limited MF lesions. To our knowledge this is the longest use of the drug by any individual. PMID- 12653925 TI - A new insight into the pathogenesis of Reiter's syndrome using bioinformatics tools. PMID- 12653926 TI - Behcet's disease and nitric oxide production. PMID- 12653927 TI - Congenital self-healing Langerhans cell histiocytosis: the need for a long term follow up. PMID- 12653928 TI - Atopy patch test reactivity to house dust mites in patients with scabies. PMID- 12653931 TI - Mitochondrial diversity evaluated by the single strand conformation polymorphism method in African and North American house flies (Musca domestica L.). AB - Single strand conformation polymorphisms (SSCPs) provide a convenient and inexpensive method of surveying mitochondrial genetic variation in large samples. We investigated how much variation should be incorporated into such surveys by scoring SSCP variation at eight mitochondrial loci in each of four sub-Saharan African and four North American house fly (Diptera: Muscidae.) populations. Hierarchical analysis of diversity was performed on haplotype frequencies at each locus and on haplotype frequencies formed by combining haplotypes from two, three, four, five and eight loci. Composite haplotypes at two loci were as informative about population structure as those composed of a greater number of loci. Increasing the number of loci increased diversity estimates within, but not between, populations. Mean composite haplotype diversities (16S2 and COII) were 0.49 +/- 0.09 among the African populations and 0.32 +/- 0.08 among the North American populations. Only two of 16 haplotypes were shared between continents. Nei's genetic differentiation statistic between populations in continents GPC was 0.30 +/- 0.06 and mean genetic differentiation between continents GCT was 0.39 +/ 0.06. We conclude that there has been little detectable gene flow between North America and sub-Saharan Africa. PMID- 12653932 TI - Development of an orally infectious Sindbis virus transducing system that efficiently disseminates and expresses green fluorescent protein in Aedes aegypti. AB - We have constructed an orally infectious Sindbis virus, ME2/5'2J/GFP, that expresses green fluorescent protein (GFP) in the midgut of Aedes aegypti and in other tissues as the virus disseminates. This virus has two unique features that are improvements over the SIN-based expression systems currently used in mosquitoes. First, a subgenomic RNA promoter and GFP coding sequence is located 5'- to the second subgenomic promoter and structural genes of the virus. Second, the E2 glycoprotein gene of TE/5'2J/GFP is replaced with the E2 gene of MRE16 SIN virus. The first feature enhances virus genome stability during virus dissemination from the midgut to other tissues and the second allows efficient virus entry into the midgut epithelial cells and then spread of the virus throughout the mosquito. PMID- 12653933 TI - Linkage and mapping analysis of a non-susceptibility gene to densovirus (nsd-2) in the silkworm, Bombyx mori. AB - Nonsusceptibility to Bombyx mori densovirus type 2 (BmDNV-2) is controlled by a recessive non-susceptibility gene, nsd-2 (non-susceptibility to DNV-2) in B. mori. Taking advantage of a lack of crossing over in females, reciprocal backcrossed F1 (BF1) progeny were used for linkage analysis and mapping of nsd-2 using silkworm strains C124 and 902, which are classified as being highly susceptible and non-susceptible to DNV-2, respectively. BF1 larvae were inoculated twice with DNV-2 virus at the first and second instar stages. DNA was extracted from each of the surviving fifth instar larvae and analysed by RFLP inheritance patterns using probes specific to each of the 28 linkage groups of B. mori. Our results indicated that the non-susceptibility gene was linked to linkage group 17, since all surviving larvae showed the homozygous profile of strain 902 in their genotype. The other linkage groups showed mixtures of heterozygous and homozygous genotypes, indicating an independent assortment. A linkage map of 30.6 cM was constructed for linkage group 17 with nsd-2 mapped at 24.5 cM and three closely linked cDNA markers were identified. PMID- 12653934 TI - Chemosensory proteins of Locusta migratoria. AB - Two different classes of chemosensory proteins (CSPs) in Locusta migratoria have been identified on the basis of the molecular cloning of a series of different cDNAs from the antennae of this insect. Several CSP isoforms have been purified and biochemically characterized from antennal and wing extracts, some of them corresponding to expression products predicted for the identified cDNAs. In wings, the nature of the main endogenous ligand binding to these proteins was determined as oleoamide by a gas chromatography-mass spectrometric approach. One of these isoforms has been expressed in a bacterial system with high yield and used in a fluorescent binding assay. Competitive binding experiments have indicated the presence of long-chain compounds among the best ligands. PMID- 12653935 TI - Cowpea bruchid Callosobruchus maculatus uses a three-component strategy to overcome a plant defensive cysteine protease inhibitor. AB - The soybean cysteine protease inhibitor, soyacystatin N (scN), negatively impacts growth and development of the cowpea bruchid, Callosobruchus maculatus[Koiwa et al. (1998) Plant J 14: 371-379]. However, the developmental delay and feeding inhibition caused by dietary scN occurred only during the early developmental stages (the 1st, 2nd and 3rd instars) of the cowpea bruchid. The 4th instar larvae reared on scN diet (adapted) exhibited rates of feeding and development which were comparable to those feeding on an scN-free diet (unadapted) prior to pupation. Total gut proteolytic capacity at this larval stage significantly increased in the scN-adapted insects. The elevated enzymatic activity was attributed to a differential expression of insect gut cysteine proteases (representing the major digestive enzymes), and of aspartic proteases. scN degradation by the gut extract was observed only in adapted bruchids, and this activity appeared to be a combined effect of scN-induced cysteine and aspartic proteases. Thirty cDNAs encoding cathepsin L-like cysteine proteases were isolated from insect guts, and they were differentially regulated by dietary scN. Our results suggest that the cowpea bruchid adapts to the challenge of scN by qualitative and quantitative remodelling of its digestive protease complement, and by activating scN-degrading protease activity. PMID- 12653936 TI - Substitution of a non-retinal phospholipase C in Drosophila phototransduction. AB - The Drosophila norpA gene encodes at least two subtypes of phospholipase C (PLC), one of which is essential for phototransduction and the other is utilized in signalling pathways other than phototransduction. The two subtypes of norpA-PLC differ by 14 amino acids that have been proposed as important for the function of PLC in different signalling pathways. The present study aimed to determine whether norpA subtype II enzyme can functionally substitute for the subtype I enzyme in the phototransduction pathway. We found that the non-retinal norpA-PLC enzyme can substitute for its retinal counterpart, but that there is a reduced rate of repolarization of photoreceptors following intense light stimuli. This reduced repolarization might be due to the inability of a regulatory component being able to interact with the non-retinal norpA-PLC enzyme. PMID- 12653937 TI - Molecular evolutionary analyses of mariners and other transposable elements in fire ants (Hymenoptera: Formicidae). AB - Screens of a library of genomic DNA made during a recent study of the fire ant Solenopsis invicta revealed the presence of three distinct types of transposable elements (TEs). Two of the recovered sequences showed a high similarity to long terminal repeat (LTR) retrotransposons, while the third showed a high homology to mariner elements. To investigate the distribution and relationships of mariners in related ants, we PCR-amplified these elements from additional Solenopsis species. Phylogenetic analyses showed that they form a single group within the mauritiana subfamily that is part of a larger clade derived from hymenopteran species. We also present partial sequence data for the two LTR-retrotransposons and describe their phylogenetic affinities. PMID- 12653938 TI - Bmmar6, a second mori subfamily mariner transposon from the silkworm moth Bombyx mori. AB - A second member of the divergent mori subfamily of mariner transposons, Bmmar6, is described from the silkworm moth Bombyx mori genome. A confident consensus sequence for Bmmar6 was obtained from a single genomic copy, 17 EST sequences, and the direct sequencing of a 'family' sequence from an amplification of all full-length genomic copies. Bmmar6 is most similar to Bmmar1 in the mori subfamily, which now also includes several fly and nematode transposons. These might be viewed as a discrete family of transposons within the IS630-Tc1-mariner superfamily with a distinctive D,D37D catalytic motif, and another small divergent D,D41D clade is recognized as their sister group of transposons. PMID- 12653939 TI - Molecular cloning and expression of the dihydrofolate reductase (DHFR) gene from adult buffalo fly (Haematobia irritans exigua): effects of antifolates. AB - The folate analogues methotrexate, aminopterin and pyrimethamine were toxic when fed in a blood meal to adult buffalo flies (Haematobia irritans exigua), but aminopterin caused greater mortality than methotrexate, while trimethoprim was not toxic to adult flies. This is the first recorded instance of mortality in adult insects caused by ingestion of folate analogues. In order to investigate the mechanism of this toxicity, the dihydrofolate reductase (DHFR) gene was cloned from adult buffalo fly cDNA using a PCR-based approach. The full-length DHFR coding sequence (BF-DHFR) was 887 bp and contained an open reading frame encoding a protein of 188 amino acids. The deduced protein sequence identities between BF-DHFR and the other known insect DHFR sequences were: Drosophila melanogaster, 75%; Aedes albopictus, 54%; Heliothis virescens, 43%. The BF-DHFR gene has a single 52 bp intron, an organization more similar to Dipteran species (Drosophila and Aedes). The cDNA encoding BF-DHFR was inserted into an Escherichia coli expression vector and the recombinant protein was expressed to levels representing about 25% of total cell protein. The active enzyme was purified by affinity chromatography on methotrexate-agarose and displayed a relatively low affinity (IC50 = 30 nm) for methotrexate. PMID- 12653940 TI - Dynamics of Rickettsia-tick interactions: identification and characterization of differentially expressed mRNAs in uninfected and infected Dermacentor variabilis. AB - To begin to explore the molecular dynamics of rickettsial tick interaction, subtractive hybridization was used to screen mRNAs in Rickettsia montanensis infected and uninfected Dermacentor variabilis. We isolated 30 cDNA fragments, 22 of which were up-regulated and eight were down-regulated in response to rickettsial infection. Based on a putative identity of 11 cDNA fragments with similarity to known protein families, the tick genetic factors have been assigned into three groups including, the tick immune response factors (alpha-2 macroglobulin and IgE-dependent histamine release factor), the receptor/adhesion molecules (the signal transducer and activator of transcription-1/3 protein inhibitor, the clathrin adaptor protein and tetraspanin) and the stress response proteins (aldose reductase, glutathione-S transferase, ferritin, nucleosome assembly protein and cyclin A protein). Density analyses of semiquantitative RT PCR amplified products demonstrated differential expression for 18 of the 23 tested genetic factors, apparently representing a 78% agreement with results obtained by subtractive hybridization. Additionally, mRNA transcripts of 17 of the 23 tested genetic factors were amplified from tick haemocytes/circulatory cells demonstrating that their expression is not restricted to the ovaries and suggesting a potential involvement in the immune response. PMID- 12653941 TI - Amplifying whole insect genomes with multiple displacement amplification. AB - Many applications in insect genetics require repeated analyses on individual organisms. These include population genetic and genomics, linkage mapping, molecular systematics and map-based positional cloning. However, using the polymerase chain reaction, a limited number of analyses are possible with DNA isolated from whole bodies or parts of small or preserved specimens. We describe the optimization of a new technique, Multiple Displacement Amplification, for 100 400-fold amplification of whole mosquito genomes. We demonstrate that MDA amplifies genomic DNA directly from an adult leg or from organisms as small as a first instar mosquito larva. Genetic polymorphisms revealed at individual loci are the same whether using the original genomic DNA or MDA DNA as a template. PMID- 12653942 TI - A preliminary linkage map of the hard tick, Ixodes scapularis. AB - A linkage map of the Ixodes scapularis genome was constructed, based upon segregation amongst 127 loci. These included 84 random amplified polymorphic DNA (RAPD) markers, 32 Sequence-Tagged RAPD (STAR) markers, 5 cDNAs, and 5 microsatellites in 232 F1 intercross progeny from a single, field-collected P1 female. A preliminary linkage map of 616 cM was generated across 14 linkage groups with one marker every 10.8 cM. Assuming a genome size of approximately 109 bp, the relationship of physical to genetic distance was found to be approximately 300 kb/cM in the I. scapularis genome. PMID- 12653943 TI - Differential diagnosis of skin lesions after allogeneic haematopoietic stem cell transplantation. AB - Allogeneic haematopoietic stem cell transplantation (i.e. bone marrow or peripheral blood stem cell transplantation) is a common procedure in the treatment of various haematological disorders such as aplastic anaemia, (pre)leukaemias, some malignant lymphomas, multiple myeloma and immunodeficiency states. Many of these patients develop erythematous skin lesions following transplantation. Although graft- versus-host disease is the major differential diagnosis in these situations, many other causes of erythema are encountered. The large number of transplant patients means that more and more pathologists are confronted with the challenging problem of making a correct diagnosis in these situations. In this review article we therefore describe the different causes of erythema and their differential diagnoses. In most cases the clinical presentation is related to the microscopical features. Besides acute and chronic graft-versus-host disease, we discuss the (common) drug reactions and non specific features such as Sweet's syndrome, erythema nodosum and eosinophilic folliculitis. In addition, we deal with the recurrence of original diseases and infections. With this knowledge every pathologist should feel comfortable when looking at skin biopsies of patients after haematological stem cell transplantation. PMID- 12653944 TI - Excision biopsy findings of patients with breast needle core biopsies reported as suspicious of malignancy (B4) or lesion of uncertain malignant potential (B3). AB - AIMS: The UK National Health Service Breast Screening Programme has proposed five categories for reporting breast needle core biopsies. The majority of cores are reported as benign (B1), normal (B2) or malignant (B5). The predictive value of the two borderline categories suspicious of malignancy (B4) and lesion of uncertain malignant potential (B3) was studied. METHODS AND RESULTS: Over a 2 year period a total of 3822 breast needle core biopsies were performed, with 2997 from symptomatic patients and 825 from women undergoing mammographic screening, including 43 B4 reports (40 patients) and 120 B3 reports (116 lesions in 115 patients). The frequencies of B4 (2.5% versus 0.7%) and B3 cores (7.3% versus 2.0%) were both higher in screening than in symptomatic patients. B4 was most commonly used for small fragments of atypical cells separate from the main core or focal atypical intraductal proliferations. The criteria for calling a core B3 were: atypical intraductal epithelial proliferations (including foci that in excision specimens would be classified as atypical ductal hyperplasia), lobular neoplasia, radial scar, papillary lesion, fibroepithelial lesion with cellular stroma and spindle cell proliferations. Excision biopsies were performed in 39 patients with B4 core and 96 with B3 core. Invasive carcinoma or ductal carcinoma in situ was seen in 33 of the patients with B4 (85%) and in 29 of those with B3 cores (25%). Some categories of B3 core were associated with a higher rate of malignancy (40% for atypical intraductal epithelial proliferations and 46% for lobular neoplasia). CONCLUSIONS: The positive predictive value for carcinoma is high following a B4 core (86%). The lesion should be excised, but definitive cancer treatment is not appropriate. In some cases a definite diagnosis of malignancy can be made on repeat core. The B3 group is more heterogeneous and has a lower rate of malignancy on further biopsy (25%). The majority of B3 lesions require excision. All these patients should be discussed at multidisciplinary meetings. PMID- 12653945 TI - Calibration of immunohistochemistry for assessment of HER2 in breast cancer: results of the French multicentre GEFPICS study. AB - AIMS: HER2 protein is over-expressed in 15-30% of breast carcinomas. Immunohistochemistry (IHC) is a common and inexpensive method able to specifically detect HER2 protein. However, lack of standardization of IHC has been considered responsible for discrepancies in HER2 status assessment performed by IHC and fluorescence in-situ hybridization (FISH). This prompted us to perform a multicentric IHC calibration test to achieve a maximum accuracy of HER2-IHC compared with HER2-FISH taken as the reference method. METHODS AND RESULTS: Twelve French laboratories participated in this study, including 119 cases of invasive breast carcinomas for which both fixed and frozen tissues were available. HER2 expression was determined in fixed tissues by individual in-house IHC techniques, using either CB11 (Novocastra, Newcastle, UK) or A0485 (Dako, Glostrup, Denmark) anti-HER2 antibodies. Two cut-off values were used: 10% and 60% of immunostained cells. In 116 of the 119 cases, HER2 gene status could also be determined by FISH on frozen sections, performed in a single laboratory. Results were centralized and compared. When suboptimal concordance between IHC and FISH was observed, IHC was calibrated and a second run was performed. The specificity, sensitivity and accuracy of IHC compared with FISH were noted before and after calibration. Forty-four out of 116 (38%) tumours showed HER2 gene amplification. Accuracy of IHC was complete in the first run for 6/12 laboratories. Calibration, necessary for the six others, relied mainly on the combination of a heat-induced epitope retrieval step with an increase of dilution of the primary antibody. In the second run, HER2 over-expression was found in 46 (40%) and 44 (38%) of the 116 cases, using 10% or 60% of stained cells as cut offs, respectively. The corresponding accuracy rates were 93% and 95%. CONCLUSIONS: This study showed that a high accuracy of IHC could be obtained for the determination of HER2 status in all laboratories using their in-house IHC technique, provided that a calibration process was performed. Antigen retrieval procedure, high dilutions of anti-HER2 antibody and the use of specific controls were crucial for HER2-IHC calibration. A 95% accuracy rate of IHC, using FISH as gold standard, was obtained by considering immunolabelling HER2-IHC results as a continuous variable, and taking 60% invasive stained cells as the cut-off for HER2 over-expression. PMID- 12653946 TI - Expression of HER-2/neu gene and protein in salivary duct carcinomas of parotid gland as revealed by fluorescence in-situ hybridization and immunohistochemistry. AB - AIMS: Salivary duct carcinoma is a highly malignant salivary gland tumour with aggressive clinical behaviour, characterized by histological resemblance to invasive ductal carcinoma of the breast. Amplification of HER-2/neu oncogene and over-expression of its gene product have both prognostic and therapeutic implications in breast cancer. Recent report on salivary duct carcinomas for HER 2/neu using immunohistochemistry (IHC) has shown over-expression in most cases. However, correlation between IHC and molecular genetic analysis of HER-2/neu in salivary duct carcinoma has not yet been performed. METHODS AND RESULTS: We have now evaluated 11 cases of salivary duct carcinomas for HER-2/neu status using IHC and fluorescent in-situ hybridization (FISH). To our knowledge, this is the first molecular genetic analysis of HER-2/neu in salivary duct carcinoma. CONCLUSIONS: In immunohistochemistry, over-expression of HER-2/neu protein was identified as distinct membrane staining in most carcinoma cells in all our salivary duct carcinoma cases, while only four cases revealed an amplification of HER-2/neu gene by means of FISH analysis. Both amplified and non-amplified salivary duct carcinomas with strong immunohistochemical staining for HER-2/neu protein were associated with poor clinical outcome for the patients. Apparently, HER-2/neu protein over-expression could also be controlled by mechanisms other than gene amplification. In the group of salivary gland tumours other than salivary duct carcinoma, strong over-expression was detected only in three cases of carcinoma ex pleomorphic adenoma. Thus, over-expression of HER-2/neu protein is also a useful marker of malignant transformation in pleomorphic adenomas. PMID- 12653947 TI - Cellular differentiation status of epithelial polyps of the colorectum: the gastric foveolar cell-type in hyperplastic polyps. AB - AIMS: The 'metaplastic' polyp of the colorectum, a synonym for the hyperplastic polyp, was named based only on features of the crypt epithelium. It is considered non-neoplastic, but the precise cellular differentiation status remains to be proven. METHODS AND RESULTS: Forty-eight hyperplastic polyps, 12 serrated adenomas, 45 tubular adenomas and five juvenile polyps were studied for their phenotypic expression using gastric (foveolar or pyloric gland cell), small intestinal (goblet cell), and colonic (goblet cell) cellular markers by immunohistochemical and mucin histochemical techniques. Gastric foveolar cell type differentiation was significantly expressed in hyperplastic polyps, while colonic differentiation was also consistently preserved. Neither gastric pyloric type nor small intestinal differentiation was observed. The same cell differentiation status as hyperplastic polyps was observed in serrated adenomas but not in tubular adenomas or juvenile polyps. CONCLUSIONS: A large proportion of hyperplastic polyps are composed of hybrid epithelium, with bidirectional differentiation to both gastric foveolar and colonic epithelial cells in the same crypt. Therefore hyperplastic polyps might be interpreted as the outcome of abnormal cell differentiation of stem cells. The same phenotypic expression suggests that hyperplastic polyps and serrated adenomas share the same cell lineage. PMID- 12653948 TI - Pathological significance of oxidative cellular damage in human alcoholic liver disease. AB - AIMS: To investigate the pathological significance of oxidative stress-induced lipid peroxidation and oxidative DNA damage in alcoholic liver disease. METHODS AND RESULTS: Hepatic expression of 4-hydroxy-2'-nonenal (HNE) adducts and 8 hydroxydeoxyguanosine (8-OHdG) as reliable markers of lipid peroxidation and oxidative DNA damage, respectively, was analysed immunohistochemically and compared with histological findings in alcoholic liver disease. While no HNE adducts were observed in control livers, HNE adducts were frequently (37 of 40 cases, 92.5%) detected in alcoholic liver disease. The localization of HNE adducts was the cytoplasm of hepatocytes and sinusoidal cells in zone 3. As for 8 OHdG, 29 of 40 cases (72.5%) with alcoholic liver disease exhibited positive immunolabelling for 8-OHdG, while 8-OHdG expression was not evident in control livers. The nuclear expression of 8-OHdG was mainly detected in the hepatocytes within the areas of active inflammation. Among histological parameters, the grade of necro-inflammation activity as well as the presence of Mallory bodies were significantly associated with the expression of HNE adducts and 8-OHdG. In addition, the severity of steatosis also correlated with HNE adduct expression. CONCLUSIONS: Lipid peroxidation and oxidative DNA damage occur widely and may be associated with certain pathological features in human alcoholic liver disease. PMID- 12653949 TI - Concordance of FIGO grade of endometrial adenocarcinomas in biopsy and hysterectomy specimens. AB - AIMS: We compared the FIGO grade of endometrial adenocarcinomas in endometrial samples and subsequent hysterectomy specimens, between 1997 and 2000 in a general pathology department. Our aims were to assess concordance, and the influence of the sampling technique, patient age, reporting pathologist and interval between the biopsy and definitive surgery. In order that the study reflected working practice, the tumour grade given by the reporting pathologist was used whenever possible. METHODS AND RESULTS: Endometrial samples and the subsequent hysterectomy specimens from 125 patients were studied. The comparative FIGO grades were analysed with reference to biopsy type, patient age, reporting pathologist and interval between biopsy and hysterectomy. Concordance was 45% for grade 1, 63.3% for grade 2 and 75.6% for grade 3 carcinomas, the overall concordance being 64.5%. The concordance for grade 3 tumours was significantly higher than for grade 1. The predictive accuracy was independent of biopsy type, patient age, the interval between biopsy and hysterectomy, and whether the same pathologist reported both specimens. CONCLUSIONS: Irrespective of the sampling device used, patient age, time-lapse between biopsy and hysterectomy, or whether the same pathologist reported both specimens, the grade of endometrial adenocarcinoma often differs in the initial biopsy sample from that in the final hysterectomy specimen. This may have important implications for patient management, especially for tumours that yield low-grade biopsies. PMID- 12653950 TI - Leiomyoblastoma of the uterus: an immunohistochemical and electron microscopic study of distinctive tumours with immature smooth muscle cell differentiation mimicking fetal uterine myocytes. AB - AIMS: We present three distinctive uterine tumours which exhibited immature smooth muscle differentiation mimicking smooth muscle cells of the fetal uterus. METHODS AND RESULTS: The patients were 45, 46 and 49 years old, and all of them had simple hysterectomies. Grossly, all tumours were present in the uterine body, and two of the three tumours were well demarcated 60-mm and 85-mm lesions, and the other tumour was a small 25-mm incidental lesion within multiple conventional leiomyomas. The tumours had varied histological features and were composed of round epithelioid, rhabdoid and large vacuolated cells intermingled with spindle shaped cells to various degrees. Although their round vesicular nuclei showed mild to moderate variation in size, prominent nuclear atypia was not seen. Necrosis and mitotic figures suggesting biological aggressiveness were not present in any of the tumours. Immunohistochemically, tumour cells were intensely positive for desmin and alpha-smooth muscle actin, whereas positivity for heavy molecular weight caldesmon was restricted. In addition, two cases were positive for non-muscle myosin heavy chain (SMemb). Ultrastructurally, most tumour cells contained various amounts of intermediate filaments which were occasionally abundant and aggregated as in rhabdoid cells. Well-developed myofilaments with focal densities were observed in only a few tumour cells. Intermediate filaments and bundles of thin filaments without dense bodies were often intermingled and they occasionally formed distinctive complexes with many irregular dense body like structures and crystalloid bodies. Other cytoplasmic organelles including rather rich mitochondria, some rough endoplasmic reticulum and free ribosomes were also common. CONCLUSIONS: These findings support their immature smooth muscle cell differentiation which mimics the mesenchymal cells of fetal uterus during 14-26 weeks of gestation. The term 'uterine leiomyoblastoma' is thought to be appropriate for describing these distinctive immature smooth muscle tumours. PMID- 12653951 TI - Differential expression of S100 calcium-binding proteins in epidermoid cysts, branchial cysts, craniopharyngiomas and cholesteatomas. AB - AIMS: To investigate whether epidermoid cysts, branchial cysts, craniopharyngiomas and cholesteatomas express S100 proteins differentially by immunohistochemical assaying the presence of S100A1, S100A2, S100A3, S100A4, S100A5, S100A6 and S100B. METHODS AND RESULTS: Immunopositivity/negativity was recorded for each S100 protein in a series of 52 cases consisting of 12 epidermoid cysts, 12 branchial cysts, 15 adamantinomatous craniopharyngiomas and 13 acquired cholesteatomas. Except in the case of the craniopharyngiomas, immunoreactivity was assessed independently in the basal membrane and the basal, the internal and the keratin layers. Our data show that in contrast to S100B, which was rarely expressed, S100A1, S100A2, S100A4 and S100A5 were often present in these four types of epithelial lesions. S100A3 and S100A6 and, to a lesser extent, S100A5 were the most differentially expressed proteins across the different histopathological groups analysed. These three proteins are expressed more often in craniopharyngiomas and cholesteatomas, the two more aggressive types of lesions. CONCLUSIONS: This is the first study to report data on the expression of seven S100 proteins in different histopathological groups of epithelial head and neck lesions, whose precise embryological origins are still a matter of debate. S100 proteins could possibly be used as markers to target this embryonic origin, since our results show that S100A3 and S100A6 (and, to a lesser extent, S100A5) are expressed differentially across these different groups of epithelial lesions. PMID- 12653953 TI - Adenocarcinoma in Crohn's disease. PMID- 12653952 TI - Expression of cyclin A and topoisomerase IIalpha of oligodendrogliomas is correlated with tumour grade, MIB-1 labelling index and survival. AB - AIMS: This study was designed to investigate immunoexpression of cyclin A and D1, and topoisomerase IIalpha in oligodendrogliomas and to evaluate the correlation with MIB-1 (Ki67), tumour grade, and survival of the patients. METHODS AND RESULTS: Forty cases of oligodendrogliomas (20 high- and 20 low-grade) were studied immunohistochemically with the above-mentioned monoclonal antibodies. RESULTS: Normal brain tissues included in tumour sections did not express any of cyclin A, MIB-1 and topoisomerase IIalpha except cyclin D1, which was shown in perineuronal and interfascicular normal oligodendroglial cells. In low-grade and high-grade oligodendrogliomas, the mean cyclin A labelling index (LI) was 1.18 +/ 0.98% versus 4.65 +/- 1.99%, respectively; the mean topoisomerase IIalpha LI was 1.32 +/- 1.04% versus 6.63 +/- 4.31%, respectively; and the mean MIB-1 LI was 1.69 +/- 1.55% versus 9.46 +/- 4.66%, respectively. Interestingly, cyclin D1 was not expressed in any oligodendrogliomas. Both cyclin A and topoisomerase IIalpha LI showed a significant positive correlation with MIB-1 LI and histological grade of oligodendrogliomas (P < 0.01) and an inverse correlation with overall survival (P < 0.01). Univariate analysis showed that cyclin A and topoisomerase IIalpha LIs with a cut-off point at 3% were a significant prognostic factor (P: cyclin A = 0.0040, topoisomerase IIalpha = 0.0033). CONCLUSION: Cyclin A and topoisomerase IIalpha expression are closely correlated with anaplastic oligodendrogliomas and worse clinical outcomes. Cyclin D1 seems not to be involved in the tumorigenesis of oligodendrogliomas. PMID- 12653954 TI - Intestinal polyposis due to multiple myeloma. PMID- 12653955 TI - CD30 expression in L&H cells of Hodgkin's disease, nodular lymphocyte predominant type. PMID- 12653956 TI - The origin of osteoclast-like giant cells in atypical fibroxanthoma. PMID- 12653957 TI - Intra-abdominal small round cell tumour with EWS-WT1 fusion transcript in an elderly patient. PMID- 12653959 TI - Tamo selectively modulates nuclear import in Drosophila. AB - BACKGROUND: The NF-kappaB/Rel pathway functions in the establishment of dorsal ventral polarity and in the innate humoral and cellular immune response in Drosophila. An important aspect of all NF-kappaB/Rel pathways is the translocation of the Rel proteins from the cytoplasm to the nucleus, where they function as transcription factors. RESULTS: We have identified a new protein, Tamo, which binds to Drosophila Rel protein Dorsal, but not to Dorsal lacking the nuclear localization sequence. Tamo does not bind to the other Drosophila Rel proteins, Dif and Relish. The Tamo-Dorsal complex forms in the cytoplasm and Tamo also interacts with a cytoplasmically orientated nucleoporin. In addition Tamo binds the Ras family small GTPase, Ran. Tamo functions during oogenesis and, based on phenotypic analysis, controls the levels of nuclear Dorsal in early embryos. It further regulates the accumulation of Dorsal in the nucleus after immune challenge. CONCLUSIONS: Tamo has an essential function during oogenesis. Tamo interacts with Dorsal and proteins that are part of the nuclear import machinery. We propose that tamo modulates the levels of import of Dorsal and other proteins. PMID- 12653960 TI - The second phase activation of protein kinase C delta at late G1 is required for DNA synthesis in serum-induced cell cycle progression. AB - BACKGROUND: Cell lines that stably over-express protein kinase C (PKC) delta frequently show a decrease in growth rate and saturation density, leading to the hypothesis that PKC delta has a negative effect on cell proliferation. However, the mode of PKC delta activation, the cell cycle stage requiring PKC delta activity, and the exact role of PKC delta at that stage remains unknown. RESULTS: Here we show that the treatment of quiescent fibroblasts with serum activates PKC delta at two distinct time points, within 10 min after serum treatment, and for a longer duration between 6 and 10 h. This biphasic activation correlates with the phosphorylation of Thr-505 at the activation loop of PKC delta. Importantly, an inhibitor of PKC delta, rottlerin, suppresses the biphasic activation of PKC delta, and suppression of the second phase of PKC delta activation is sufficient for the suppression of DNA synthesis. Consistent with this, the transient over expression of PKC delta mutant molecules lacking kinase activity suppresses serum induced DNA synthesis. These results imply that PKC delta plays a positive role in cell cycle progression. While the over-expression of PKC delta enhances serum induced DNA synthesis, this was not observed for PKC epsilon. Similar experiments using a series of PKCdelta/ epsilon chimeras showed that the carboxyl-terminal 51 amino acids of PKC delta are responsible for the stimulatory effect. On the other hand, the over-expression of PKC delta suppresses cell entry into M-phase, being consistent with the previous studies based on stable over-expressors. CONCLUSIONS: We conclude that PKC delta plays a role in the late-G1 phase through the positive regulation of cell-cycle progression, in addition to negative regulation of the entry into M-phase. PMID- 12653961 TI - A SWI2/SNF2-type ATPase/helicase protein, mDomino, interacts with myeloid zinc finger protein 2A (MZF-2A) to regulate its transcriptional activity. AB - BACKGROUND: The myeloid zinc finger protein 2A (MZF-2A) is a Kruppel-type C2H2 zinc finger transcription factor expressed in myeloid cells and involved in the growth, differentiation and tumorigenesis of myeloid progenitors. Previously we identified a 180 amino acid domain in MZF-2A which is responsible for the transcriptional activation of MZF-2A. To understand the mechanism of the MZF-2A dependent transcriptional activation, we screened for molecules that interact with the transactivation domain (TAD) of MZF-2A. RESULTS: By using the yeast Ras recruitment two-hybrid screening, we identified a novel SWI2/SNF2-related protein, termed mammalian Domino (mDomino), as an MZF-2A-binding partner. The mDomino protein, which shows a marked similarity to the Drosophila Domino protein, contains a SWI2/SNF2-type ATPase/helicase domain, a SANT domain, and a glutamine-rich (Q-rich) domain. The C-terminal Q-rich domain of mDomino physically associates with the TAD of MZF-2A in mammalian cells as well as in yeast. Expression of the mDomino Q-rich domain, together with MZF-2A in myeloid LGM-1 cells, enhanced the MZF-2A-mediated activation of a reporter gene. CONCLUSIONS: These results strongly suggest that an ATP-dependent chromatin remodelling complex containing mDomino interacts with MZF-2A to regulate gene expression in myeloid cells. PMID- 12653962 TI - A brute force postgenome approach to identify temperature-sensitive mutations that negatively interact with separase and securin plasmids. AB - BACKGROUND: The fission yeast Schizosaccharomyces pombe separase/Cut1 and securin/Cut2 are required for anaphase-specific activation of proteolysis that leads to proper sister chromatid separation. We intended to identify ts (temperature sensitive) strains whose growth was inhibited by multicopy plasmid pCUT1 or pCUT2 at the permissive temperature. RESULTS: After a one-by-one transformation of 1015 randomly isolated ts strains, 18 transformants that retarded in colony formation at the permissive or semipermissive temperature were isolated. Six of them, in the absence of pCUT1 or pCUT2, produced mitotic phenotypes with condensed chromosomes at the restrictive temperature. Gene cloning established that these mutants were defective in either the subunits (Cut9, Cut23, Cut20 or Apc10) of APC (anaphase promoting complex)/cyclosome or Cut8, a regulator for 26S proteasome localization. The inhibitory effect of separase against APC/cyclosome mutations was abolished when the catalytic site mutation C1730A was introduced and overproduced, indicating that inhibition needs an active separase. Securin/Cut2 overproduction also caused a negative effect on these mutants. Surprisingly, the phenotypes of cut9 and cut23 in the presence of pCUT1 or pCUT2 were not the mitotic arrest, and they were strikingly different depending on pCUT1 or pCUT2. CONCLUSIONS: This study shows the functional link between separase/Cut1 and APC/cyclosome in a separase activity-dependent manner. The negative effect of active separase overproduction on APC/cyclosome mutations is possibly due to the direct inhibition of APC/cyclosome. In addition, the manner of the inhibition by high copy securin and separase plasmids were quite different each other and did not result in the mitotic block. PMID- 12653963 TI - The small GTPase Rho4 is involved in controlling cell morphology and septation in fission yeast. AB - BACKGROUND: Rho family small GTPases have been shown to be involved in various cellular activities, including the organization of actin cytoskeleton in eukaryotic cells. There are six rho genes in the fission yeast Schizosaccharomyces pombe. Cdc42 is known to control the polarity of the cell. Rho1, Rho2 and Rho3 play important roles in controlling cell shape and septation. On the other hand, Rho4 and Rho5 have not yet been characterized. Here we report the function of rho4+ in fission yeast. RESULTS: Gene disruption revealed that rho4+ is not essential for cell growth. However, rho4-null cells were abnormally elongated and had multiple septa of irregular shape at 37 degrees C. In these cells, F-actin patches were randomly localized all over the cell periphery, and cytoplasmic microtubules (MTs) were misoriented. On the other hand, the exogenous expression of a constitutively active Rho4-G23V or Rho4-Q74L in wild-type cells induced depolarization of F-actin patches and cytoplasmic MTs. Rho4 was localized to the cell periphery during interphase and septum during mitosis. Both the binding of GTP and isoprenylation of its C-terminus were necessary for the localization. Furthermore, the localization of Rho4 was likely to be controlled by Rho GAP and Rho GDI. CONCLUSION: Rho4 may control cell morphogenesis and septation by regulating both the actin cytoskeleton and cytoplasmic MTs. PMID- 12653964 TI - Structure-function analysis of human Spt4: evidence that hSpt4 and hSpt5 exert their roles in transcriptional elongation as parts of the DSIF complex. AB - BACKGROUND: The human Spt4/Spt5 complex, termed DRB-sensitivity inducing factor (DSIF) is a dual regulator of transcription that stimulates, or, when cooperating with negative elongation factor (NELF), represses RNA polymerase II (RNAPII) elongation. Spt4 and Spt5 are also thought to be involved in mRNA capping, homologous DNA recombination, and transcription-coupled DNA repair. As a first step to understanding how these proteins regulate diverse cellular processes, we investigated the structure and function of hSpt4 in vitro. RESULTS: Immunodepletion of hSpt5 from HeLa nuclear extracts resulted in the efficient co depletion of hSpt4. Using DSIF-depleted nuclear extracts and a series of Spt4 mutants, we examined the amino acid sequence of hSpt4 which was important for hSpt5 binding and for transcriptional repression and activation by DSIF. Unexpectedly, the zinc finger of hSpt4, which is critical for the yeast counterpart to function in vivo, was dispensable for hSpt5 binding and for transcriptional regulation in vitro. CONCLUSION: These and other results suggest: (i) that the central region of hSpt4 is necessary and sufficient for its function in vitro and (ii) that there is no free hSpt4 or hSpt5 in cells. We propose that hSpt4 and hSpt5 exert their roles in transcriptional regulation, and possibly in other nuclear processes, as parts of the DSIF complex. PMID- 12653965 TI - Keap1 regulates both cytoplasmic-nuclear shuttling and degradation of Nrf2 in response to electrophiles. AB - BACKGROUND: Transcription factor Nrf2 regulates the expression of a set of detoxifying and anti-oxidant enzyme genes. Several lines of evidence suggest that electrophiles and reactive oxygen species liberate Nrf2 from its cytoplasmic repressor Keap1 and provoke the accumulation of Nrf2 in the nucleus. To elucidate the molecular mechanisms as to how Nrf2 is activated by inducers, we examined the cytoplasmic-nuclear shuttling and turnover of Nrf2. RESULTS: We found that Nrf2 is rapidly degraded through the proteasome pathway, while electrophiles cause Nrf2 nuclear translocation with concomitant stabilization. Crucial to the inducible accumulation of Nrf2 is the enfeebling of the Nrf2-Keap1 interaction by electrophiles. Exploiting mice which have the LacZ reporter gene knocked into the nrf2 locus, we revealed that the inducible accumulation of Nrf2 protein by electrophiles in macrophages and intestinal epithelia could be recapitulated by the Nrf2 N-terminal region in combination with a nuclear localization signal. We also found constitutive Nrf2 nuclear accumulation in Keap1-deficient mouse macrophages. CONCLUSIONS: Our results highlight the fact that Nrf2 protein turnover is regulated by Keap1 mediated subcellular compartmentalization. PMID- 12653966 TI - Enforced cytokinesis without complete nuclear division in embryonic cells depleting the activity of DNA topoisomerase IIalpha. AB - BACKGROUND: There are two distinct DNA topoisomerase II (topo II) isoforms, designated topo IIalpha and topo IIbeta, in mammalian cells. The function of topo IIalpha in the development of mammalian cells has not been elucidated because of a lack of topo IIalpha mutants. RESULTS: We generated mice with a targeted disruption of the topo IIalpha gene. The development of topo IIalpha-/- embryos was terminated at the 4- or 8-cell stage. When wild-type embryos at the 2- or 4 cell stage were treated with ICRF-193, a catalytic inhibitor of topo II, nuclear division occurred followed by cytokinesis to form 4 or 8 cells, respectively, then development was terminated. Microscope analysis of 4,6-diamidino-2 phenylindole (DAPI)-stained nuclei of both topo IIalpha-/- and ICFR-193-treated embryonic cells revealed a droplet-like structure connecting the terminals of two adjacent nuclei forming a bridge-like structure. Phosphorylated histone H3, a marker for the M phases, disappeared from the nuclei of the topo IIalpha-depleted embryonic cells. Laser scanning cytometry of the topo IIalpha-depleted cells revealed the presence of 2N DNA cells. CONCLUSIONS: Our results indicate that topo IIalpha has an essential role in the early stages of mouse development and that depletion of topo IIalpha from the embryonic cells causes incomplete nuclear division followed by enforced cytokinesis. PMID- 12653968 TI - Alpha2A and alpha2C-adrenoceptor regulation in the brain: alpha2A changes persist after chronic stress. AB - Stress-induced activation of the central nervous noradrenergic system has been suspected to induce depressive disorders. As episodes of depression often occur some time after a stress experience we investigated whether stress-induced changes in the alpha2-adrenoceptor (alpha2-AR) system persist throughout a post stress recovery period. Brains of male tree shrews were analysed after 44 days of chronic psychosocial stress and after a subsequent 10-day recovery period. Expression of RNA for alpha2A and alpha2C-adrenoceptors was quantified by in situ hybridization, and receptor binding was determined by in vitro receptor autoradiography. Activities of the sympathetic nervous system and of the hypothalamo-pituitary-adrenal axis were increased during chronic stress but normalized during recovery. Alpha2A-AR RNA in the glutamatergic neurons of the lateral reticular nucleus was elevated significantly after stress and after recovery (by 29% and 17%). In the dorsal motor nucleus of the vagus, subtype A expression was enhanced after recovery (by 33%). In the locus coeruleus, subtype A autoreceptor expression was not changed significantly. Subtype C expression in the caudate nucleus and putamen was elevated by stress (by 5 and 4%, respectively) but normalized during recovery. Quantification of 3H-RX821002 binding revealed receptor upregulation during stress and/or recovery. Our data therefore show: (i) that chronic psychosocial stress differentially regulates expression of alpha2-adrenoceptor subtypes A and C; (ii) that subtype A heteroreceptor expression is persistently upregulated whereas (iii), subtype C upregulation is only transient. The present findings coincide with post mortem studies in depressed patients revealing upregulation of alpha2A-ARs. PMID- 12653967 TI - Alleles of the proximal promoter of BAT1, a putative anti-inflammatory gene adjacent to the TNF cluster, reduce transcription on a disease-associated MHC haplotype. AB - BACKGROUND: BAT1 belongs to the DEAD-box family of proteins, and is encoded in the central region of the MHC, a region containing genes affecting immunopathological disorders including Type 1 diabetes. We showed that BAT1 can reduce inflammatory cytokine production, supporting its candidacy as a disease gene. Here we examined the proximal promoter region of BAT1. RESULTS: Ten single nucleotide polymorphisms were identified in approximately 1.4 kb of sequence, defining at least seven alleles. Sections of the BAT1 promoter region were amplified from cells homozygous for the MHC haplotypes associated with susceptibility (HLA-A1, B8, DR3; 8.1 haplotype) and resistance (HLA-A3, B7, DR15; 7.1 haplotype) to diabetes and cloned into a promoter-less luciferase-encoding plasmid. Jurkat cells transiently transfected with fragments from the 8.1 haplotype exhibited a lower luciferase activity than those transfected with fragments from the 7.1 haplotype, indicating reduced transcription. The effect was clearest with the 520 bp immediately upstream of the transcriptional start site. Electrophoretic mobility shift assays using oligonucleotides spanning polymorphic sites within the 520 bp (proximal) promoter fragment showed haplotype specific binding of nuclear proteins. CONCLUSIONS: In view of the anti inflammatory role of BAT1, reduced production on a disease-associated haplotype constitutes a novel and self-consistent model for the effect of central MHC genes on disease. PMID- 12653969 TI - Aberrant responses to acoustic stimuli in mice deficient for neural recognition molecule NB-2. AB - NB-2, a member of the contactin subgroup in the immunoglobulin superfamily, is expressed specifically in the postnatal nervous system, reaching a maximum level at 3 weeks postnatal. NB-2 displays neurite outgrowth-promoting activity in vitro. To assess its function in the nervous system, we generated mutant mice in which a part of the NB-2 gene was ablated and replaced with the tau-LacZ gene. The general appearance of NB-2-deficient mice and their gross anatomical features were normal. The LacZ expression patterns in heterozygous mice revealed that NB-2 is preferentially expressed in the central auditory pathways. In the audiogenic seizure test, NB-2-deficient mice exhibited a lower incidence of wild running, but a higher mortality rate than the wild-type littermates. c-Fos immunohistochemistry demonstrated that neural excitability induced by the audiogenic seizure test in the NB-2-deficient mice was prominently attenuated in both the dorsal and external cortices of the inferior colliculus, where enhanced neural excitability was observed in the wild-type mice. In response to pure-tone stimulation after priming, NB-2-deficient mice exhibited a diffuse and low level of c-Fos expression in the central nucleus of the inferior colliculus, which was distinctly different from the band-like c-Fos expression corresponding to the tonotopic map in the wild-type littermates. Taken together, these results suggest that a lack of NB-2 causes impairment of the neuronal activity in the auditory system. PMID- 12653970 TI - Actions of neuropeptide Y and growth hormone secretagogues in the arcuate nucleus and ventromedial hypothalamic nucleus. AB - Systemic or central administration of growth-hormone secretagogues (GHS) induces dense Fos expression in the arcuate nucleus but little or no Fos expression in the ventromedial hypothalamic nucleus, although both sites show intense expression of mRNA for the GHS receptor. Here, we recorded the electrical activity of single neurons from the arcuate nucleus and from the ventromedial hypothalamic nucleus in a rat hypothalamic slice preparation, and compared responses of these two populations to GHS. At both sites, the predominant neuronal response to GHS was a long-lasting excitation, indicating that GHS receptors at both sites are functional and similarly coupled to electrical excitation. We also tested neurons at both sites for their responses to neuropeptide Y and to somatostatin; at both sites the predominant response to each of these peptides was inhibitory. The arcuate cells that are activated by GHS include neuropeptide Y cells and growth hormone-releasing hormone cells. It seems possible that neuropeptide Y released in the ventromedial hypothalamus from the terminals of arcuate neurons counteracts the activation of ventromedial hypothalamic neurons by GHS in vivo, or that somatostatin released following liberation of growth hormone may do so. PMID- 12653971 TI - Stage-specific activity patterns affect motoneuron axonal retraction and outgrowth during the metamorphosis of Manduca sexta. AB - During the metamorphosis of holometabolous insects, most larval muscles and sensory neurons are replaced by new adult elements, whereas most motoneurons persist and are remodelled to serve new adult functions. In Manduca sexta, the formation of the anlagen of the adult dorsal longitudinal flight muscle (DLM) is characterized by retraction of axonal terminals and dendrites of persisting larval motoneurons, partial target muscle degeneration and myoblast accumulation during late larval life. Most of these structural changes have been attributed to hormonal control, not only because ecdysteroids govern metamorphosis, but also because motoneurons express ecdysteroid receptors and experimental manipulations of ecdysteroid titres perturb normal development. To test whether activity dependent mechanisms also came into play, chronic extracellular recordings were conducted in vivo from the five future DLM motoneurons throughout the last 3 days of larval life. Motoneuron activity is regulated developmentally. The types of motoneurons recruited, the number of motor spikes and the duration of bursts change in a stereotypical fashion during different stages, indicating an internal control of motor activity. A characteristic cessation in the activity of the five future DLM motoneurons coincides in time with the retraction of their dendrites and their terminal arborizations, whereas their activation during ecdysis coincides with the onset of new outgrowth. Inducing advanced activity by stimulating the motoneurons selectively with ecdysis-like patterns results in significant outgrowth of their terminal arborizations. Therefore, steroids might act in concert with activity-dependent mechanisms during the postembryonic modifications of neuromuscular systems. PMID- 12653972 TI - The expression of genes encoding visual components is regulated by a circadian clock, light environment and age in the honeybee (Apis mellifera). AB - The honeybee, Apis mellifera, has been used as a model to study the development of the visual system and adult bee behaviour. However, the regulation of the levels of visual component genes has never been addressed in this organism. We isolated honeybee cDNAs encoding green-sensitive opsin and visual arrestin and then measured their mRNA levels in honeybee workers. Both mRNAs fluctuate on a daily cycle that depends on a pacemaker that functions separately from the pacemaker which controls rhythmic locomotor activity. The cycling-patterns of opsin and arrestin mRNAs are different from each other and are modified by light. Furthermore, light exposure can increase the absolute levels of both mRNAs and the arrestin mRNA level is also dependent on age. Consistent with these results, both mRNA levels are higher in foragers than in in-hive bees under natural conditions. This study thus shows that the expression of genes encoding visual components is regulated by multiple factors and is adjusted to the honeybees' need for vision during the day, and throughout their lives. Comparison of data obtained with honeybees and other organisms indicates that there is a link between the regulation of phototransduction components and vision-related animal behaviour. PMID- 12653973 TI - Regulation of RGS proteins by chronic morphine in rat locus coeruleus. AB - The present study explored a possible role for RGS (regulators of G protein signalling) proteins in the long term actions of morphine in the locus coeruleus (LC), a brainstem region implicated in opiate physical dependence and withdrawal. Morphine influences LC neurons through activation of micro -opioid receptors, which, being Gi/o-linked, would be expected to be modulated by RGS proteins. We focused on several RGS subtypes that are known to be expressed in this brain region. Levels of mRNAs encoding RGS2, -3, -4, -5, -7, -8 and -11 are unchanged following chronic morphine, but RGS2 and -4 mRNA levels are increased 2-3-fold 6 h following precipitation of opiate withdrawal. The increases in RGS2 and -4 mRNA peak after 6 h of withdrawal and return to control levels by 24 h. Immunoblot analysis of RGS4 revealed a striking divergence between mRNA and protein responses in LC: protein levels are elevated twofold following chronic morphine and decrease to control values by 6 h of withdrawal. In contrast, levels of RGS7 and -11 proteins, the only other subtypes for which antibodies are available, were not altered by these treatments. Intracellular application of wild-type RGS4, but not a GTPase accelerating-deficient mutant of RGS4, into LC neurons diminished electrophysiological responses to morphine. The observed subtype- and time-specific regulation of RGS4 protein and mRNA, and the diminished morphine induced currents in the presence of elevated RGS4 protein levels, indicate that morphine induction of RGS4 could contribute to aspects of opiate tolerance and dependence displayed by LC neurons. PMID- 12653974 TI - Direct and indirect interactions between cannabinoid CB1 receptor and group II metabotropic glutamate receptor signalling in layer V pyramidal neurons from the rat prefrontal cortex. AB - At proximal synapses from layer V pyramidal neurons from the rat prefrontal cortex, activation of group II metabotropic glutamate receptors (group II mGlu) by (2S,2'R,3'R)-2-(2',3'-dicarboxycyclopropyl) glycine (DCG IV) induced a long lasting depression of excitatory postsynaptic currents. Paired-pulse experiments suggested that the depression was expressed presynaptically. Activation of type 1 cannabinoid receptors (CB1) by WIN 55,212-2 occluded the DCG IV-induced depression in a mutually occlusive manner. At the postsynaptic level, WIN 55,212 2 and DCG IV were also occlusive for the activation of extracellular signal regulated kinase. The postsynaptic localization of active extracellular signal regulated kinase was confirmed by immunocytochemistry after activation of CB1 receptors. However, phosphorylation of extracellular signal-regulated kinase in layer V pyramidal neurons was dependent on the activation of N-methyl-d-aspartate receptors, consequently to a release of glutamate in the local network. Group II mGlu were also shown to be involved in long-term changes in synaptic plasticity induced by high frequency stimulations. The group II mGlu antagonist (RS)-alpha methylserine-O-phosphate monophenyl ester (MSOPPE) favoured long-term depression. However, no interaction was found between MSOPPE, WIN 55,212-2 and the CB1 receptor antagonist SR 141716A on the modulation of long-term depression or long term potentiation and the effects of these drugs were rather additive. We suggest that CB1 receptor and group II mGlu signalling may interact through a presynaptic mechanism in the induction of a DCG IV-induced depression. Postsynaptically, an indirect interaction occurs for activation of extracellular signal-regulated kinase. However, none of these interactions seem to play a role in synaptic plasticities induced with high frequency stimulations. PMID- 12653975 TI - Delayed peripheral nerve regeneration and central nervous system collateral sprouting in leucocyte common antigen-related protein tyrosine phosphatase deficient mice. AB - Cell adhesion molecule-like receptor-type protein tyrosine phosphatases have been shown to be important for neurite outgrowth and neural development in several animal models. We have previously reported that in leucocyte common antigen related (LAR) phosphatase deficient (LAR-deltaP) mice the number and size of basal forebrain cholinergic neurons, and their innervation of the hippocampal area, is reduced. In this study we compared the sprouting response of LAR deficient and wildtype neurons in a peripheral and a central nervous system lesion model. Following sciatic nerve crush lesion, LAR-deltaP mice showed a delayed recovery of sensory, but not of motor, nerve function. In line with this, neurofilament-200 immunostaining revealed a significant reduction in the number of newly outgrowing nerve sprouts in LAR-deltaP animals. Morphometric analysis indicated decreased axonal areas in regenerating LAR-deltaP nerves when compared to wildtypes. Nonlesioned nerves in wildtype and LAR-deltaP mice did not differ regarding myelin and axon areas. Entorhinal cortex lesion resulted in collateral sprouting of septohippocampal cholinergic fibres into the dentate gyrus outer molecular layer in both genotype groups. However, LAR-deltaP mice demonstrated less increase in acetylcholinesterase density and fibre number at several time points following the lesion, indicating a delayed collateral sprouting response. Interestingly, a lesion-induced reduction in number of (septo-entorhinal) basal forebrain choline acetyltransferase-positive neurons occurred in both groups, whereas in LAR-deltaP mice the average cell body size was reduced as well. Thus, regenerative and collateral sprouting is significantly delayed in LAR-deficient mice, reflecting an important facilitative role for LAR in peripheral and central nervous system axonal outgrowth. PMID- 12653976 TI - Receptor-selective changes in mu-, delta- and kappa-opioid receptors after chronic naltrexone treatment in mice. AB - Chronic treatment with the opioid antagonist naltrexone induces functional supersensitivity to opioid agonists, which may be explained by receptor up regulation induced by opioid receptor blockade. In the present study, the levels of opioid receptor subtypes through the brain of mice were determined after chronic naltrexone treatment using quantitative in vitro autoradiography. This is the first complete mapping study in mice for micro-, delta- and kappa-opioid receptors after chronic naltrexone exposure. Treatment with naltrexone clearly induced up-regulation of micro- (mean 80%) and, to a lesser extent, delta-opioid receptors (mean 39%). The up-regulation of micro- and delta-opioid receptors was evident throughout the brain, although there was variation in the percentage change across brain regions. In contrast, consistent up-regulation of kappa opioid receptors was observed in cortical structures only and was not so marked as for micro- and delta-opioid receptors. In noncortical regions kappa-opioid receptor expression was unchanged. Taken together, the present findings suggest opioid receptor subtype-selective regulation by chronic naltrexone treatment in mice. PMID- 12653977 TI - Mechanisms underlying the noradrenergic modulation of longitudinal coordination during swimming in Xenopus laevis tadpoles. AB - Noradrenaline (NA) is a potent modulator of locomotion in many vertebrate nervous systems. When Xenopus tadpoles swim, waves of motor neuron activity alternate across the body and propagate along it with a brief rostro-caudal delay (RC delay) between segments. We have now investigated the mechanisms underlying the reduction of RC-delay s by NA. When recording from motor neurons caudal to the twelfth postotic cleft, the mid-cycle inhibition was weak and sometimes absent, compared to more rostral locations. NA enhanced and even unmasked inhibition in these caudal neurons and enhanced inhibition in rostral neurons, but to a lesser extent. Consequently, the relative increase in the amplitude of the inhibition was greater in caudal neurons, thus reducing the RC-inhibitory gradient. We next investigated whether NA might affect the electrical properties of neurons, such that enhanced inhibition under NA might promote postinhibitory rebound firing. The synaptic inputs during swimming were simulated using a sustained positive current, superimposed upon which were brief negative currents. When these conditions were held constant NA enhanced the probability of rebound firing- indicating a direct effect on membrane properties--in addition to any indirect effect of enhanced inhibition. We propose that NA preferentially enhances weak caudal inhibition, reducing the inhibitory gradient along the cord. This effect on inhibitory synaptic transmission, comprising parallel pre- and postsynaptic components, will preferentially facilitate rebound firing in caudal neurons, advancing their firing relative to more rostral neurons, whilst additionally increasing the networks ability to sustain the longer cycle periods under NA. PMID- 12653978 TI - Long-term potentiation of mGluR1 activity by depolarization-induced Homer1a in mouse cerebellar Purkinje neurons. AB - Metabotropic glutamate receptor 1 (mGluR1) plays a crucial role in synaptic plasticity and motor learning in the cerebellum. We have studied activity dependent changes in mGluR1 function in mouse cultured Purkinje neurons. Depolarizing stimulation potentiated Ca2+ and current responses to an mGluR1 agonist for several hours in the cultured Purkinje neurons. It also blocked internalization of mGluR1 and increased the number of mGluR1s on the cell membrane. We found that depolarization simultaneously increased transcription of Homer1a in Purkinje neurons. Homer1a inhibited internalization and increased cell surface expression of mGluR1 when coexpressed in human embryonic kidney (HEK)-293 cells. Depolarization-induced Homer1a expression in Purkinje neurons was blocked by a mitogen-activated protein kinase (MAPK) inhibitor. Changes in internalization and mGluR1-mediated Ca2+ response were also blocked by inhibition of MAPK activity, suggesting that localization and activity of mGluR1 were regulated in the same signalling pathway as Homer1a expression. It is thus suggested that depolarization of the Purkinje neuron leads to the increment in mGluR1 responsiveness through MAPK activity and induction of Homer1a expression, which increases active mGluR1 on the cell surface by blocking internalization of mGluR1. PMID- 12653979 TI - D2 Dopamine receptor blockade results in sprouting of DA axons in the intact animal but prevents sprouting following nigral lesions. AB - Recently it was demonstrated that sprouting of dopaminergic neurons and a microglial and astrocyte response follows both partial lesions of the substantia nigra pars compacta and blockade of the D2 dopamine receptor. We therefore studied the effects of the combination of these two treatments (lesioning and D2 dopamine receptor blockade). Haloperidol administration caused a 57% increase in dopaminergic terminal tree size (measured as terminal density per substantia nigra pars compacta neuron) and an increase of glia in the striatum. Following small to medium nigral lesions (less than 60%), terminal tree size increased by 51% on average and returned density of dopaminergic terminals to normal. In contrast, administration of haloperidol for 16 weeks following lesioning resulted in reduced dopaminergic terminal density and terminal tree size (13%), consistent with absent or impaired sprouting. Glial cell numbers increased but were less than with lesions alone. When haloperidol was administered after the striatum had been reinnervated through sprouting (16-32 weeks after lesioning), terminal tree size increased up to 150%, similar to the effect of haloperidol in normal animals. By examining the effect of administering haloperidol at varying times following a lesion, we concluded that a switch in the effect of D2 dopamine receptor blockade occurred after dopaminergic synapses began to form in the striatum. We postulate that when synapses are present, D2 dopamine receptor blockade results in increased terminal density, whereas prior to synapse formation D2 dopamine receptor blockade causes attenuation of a sprouting response. We speculate that D2 dopamine receptors located on growth cones 'push' neurites toward their targets, and blockade of these receptors could lead to attenuation of sprouting. PMID- 12653980 TI - Spreading of slow cortical rhythms to the basal ganglia output nuclei in rats with nigrostriatal lesions. AB - A high proportion of neurons in the basal ganglia display rhythmic burst firing after chronic nigrostriatal lesions. For instance, the periodic bursts exhibited by certain striatal and subthalamic nucleus neurons in 6-hydroxydopamine-lesioned rats seem to be driven by the approximately 1 Hz high-amplitude rhythm that is prevalent in the cerebral cortex of anaesthetized animals. Because the striatum and subthalamic nucleus are the main afferent structures of the substantia nigra pars reticulata, we examined the possibility that the low-frequency modulations (periodic bursts) that are evident in approximately 50% nigral pars reticulata neurons in the parkinsonian condition were also coupled to this slow cortical rhythm. By recording the frontal cortex field potential simultaneously with single-unit activity in the substantia nigra pars reticulata of anaesthetized rats, we proved the following. (i) The firing of nigral pars reticulata units from sham-lesioned rats is not coupled to the approximately 1 Hz frontal cortex slow oscillation. (ii) Approximately 50% nigral pars reticulata units from 6 hydroxydopamine-lesioned rats oscillate synchronously with the approximately 1 Hz cortical rhythm, with the cortex leading the substantia nigra by approximately 55 ms; the remaining approximately 50% nigral pars reticulata units behave as the units recorded from sham-lesioned rats. (iii) Periodic bursting in nigral pars reticulata units from 6-hydroxydopamine-lesioned rats is disrupted by episodes of desynchronization of cortical field potential activity. Our results strongly support that nigrostriatal lesions promote the spreading of low-frequency cortical rhythms to the substantia nigra pars reticulata and may be of outstanding relevance for understanding the pathophysiology of Parkinson's disease. PMID- 12653982 TI - Nucleus accumbens beta-endorphin levels are not elevated by brain stimulation reward but do increase with extinction. AB - Beta-endorphin is an endogenous opioid peptide implicated in reward processes, but the brain sites directly involved in its putative role in reward have not been identified. Here we used in vivo microdialysis in rats to study the effect of a potent reinforcer, lateral hypothalamus self-stimulation (LHSS), on the extracellular levels of beta-endorphin in the nucleus accumbens (NAS). The NAS is involved in the reinforcing effects of natural and artificial rewards, has high density of opioid receptors and is innervated by arcuate nucleus beta-endorphin neurons. LHSS had no effect on extracellular levels of beta-endorphin in the NAS. Surprisingly, extinction of the self-stimulation behaviour induced a rapid increase in NAS beta-endorphin levels. In a subsequent experiment in rats previously trained to self-administer heroin for 10 days, beta-endorphin levels also were increased during a test for extinction of the heroin-reinforced behaviour. Finally, the increase in extracellular beta-endorphin levels in the NAS was also observed during exposure to an aversive stimulus, intermittent footshock (20 min). These results indicate a possible role for increased levels of NAS beta-endorphin in the organism's adaptive response to stress and frustration. PMID- 12653981 TI - Topography of cocaine-induced gene regulation in the rat striatum: relationship to cortical inputs and role of behavioural context. AB - Psychostimulants alter gene expression in projection neurons of the striatum, and such neuroplasticity is implicated in drug addiction and dependence. Evidence indicates that excitatory inputs from the cortex and thalamus are critical for these molecular changes. In the present study, we determined the topography of cocaine-induced changes in gene expression in the rat striatum and investigated whether these molecular alterations are associated with particular cortical inputs. Acute induction of c-fos (by 25 mg/kg of cocaine), and the c-fos response and dynorphin expression after repeated cocaine treatment (25 mg/kg, 4 days) were assessed as examples for short-term and longer-term molecular changes, respectively. In addition, we examined whether these molecular effects were influenced by the behaviour performed during cocaine action (running-wheel training vs. open field). Our results demonstrate that the overall topography of cocaine-induced gene regulation in the striatum is remarkably stable. Both acute and longer-term molecular changes were maximal in caudal dorsal striatal sectors that receive convergent inputs from the medial agranular and the sensorimotor cortex. In contrast, relatively minor or no effects were found in rostral and ventral striatal sectors. However, running-wheel training under the influence of cocaine enhanced the c-fos response to a subsequent cocaine challenge selectively in parts of the caudal sensorimotor striatum. These results indicate that cocaine produces molecular adaptations preferentially in cortico-basal ganglia circuits through the sensorimotor striatum, and that some of these neuronal changes are influenced by the behaviour performed during drug exposure. PMID- 12653983 TI - Analysis of differential gene expression supports a role for amyloid precursor protein and a protein kinase C substrate (MARCKS) in long-term memory. AB - Previous work has identified the intermediate and medial part of the hyperstriatum ventrale (IMHV) as a region of the chick brain storing information acquired through the learning process of imprinting. We have examined in this brain region changes in expression of candidate genes involved in memory. Chicks were exposed to a rotating red box and the strength of their preference for it, a measure of learning, determined. Brain samples were removed approximately 24 h after training. Candidate genes whose expressions were different in IMHV samples derived from strongly imprinted chicks relative to those from chicks showing little or no learning were identified using subtractive hybridization. The translation products of two candidate genes were investigated further in samples from the left and right IMHV and from two other brain regions not previously implicated in imprinting, the left and right posterior neostriatum. One of the proteins was the amyloid precursor protein (APP), the other was myristoylated alanine rich C kinase substrate (MARCKS). In the left IMHV the levels of the two proteins increased with the strength of learning. The effects in the right IMHV were not significantly different from those in the left. There were no effects of learning in the posterior neostriatum. This is the first study to relate changes in the amounts of MARCKS and APP proteins to the strength of learning in a brain region known to be a memory store and demonstrates that the systematic identification of protein molecules involved in memory formation is possible. PMID- 12653984 TI - Rhinal-hippocampal theta coherence during declarative memory formation: interaction with gamma synchronization? AB - The hippocampus and the rhinal cortex, two substructures of the medial temporal lobe, together play a crucial role in human declarative memory formation. To investigate in detail the mechanism connecting these two structures transiently during memory formation we recorded depth EEG in epilepsy patients from within the hippocampus and the rhinal cortex. During this recording, patients performed a single-trial word list-learning paradigm with a free recall memory test following a distraction task. Rhinal-hippocampal EEG coherence and spectral power at both locations in the time interval up to 2 s after onset of word presentation were analysed in the frequency range 1-19 Hz. Successful as opposed to unsuccessful memory formation was associated with a general rhinal-hippocampal coherence enhancement, but without alterations in spectral power. Coherence increases in the theta range were correlated with the previously reported memory related changes in rhinal-hippocampal gamma phase synchronization. This correlation may suggest an interaction of the two mechanisms during declarative memory formation. While theta coherence might be associated with slowly modulated coupling related to an encoding state, rhinal-hippocampal gamma synchronization may be more closely related to actual memory processes by enabling fast coupling and decoupling of the two structures. PMID- 12653985 TI - Does luminance-contrast contribute to a saliency map for overt visual attention? AB - In natural environments, humans select a subset of visual stimuli by directing their gaze to locations attended. In previous studies it has been found that at fixation points luminance-contrast is higher than average. This led to the hypothesis that luminance-contrast makes a major contribution to a saliency map of visual overt attention, consistent with a computation of stimulus saliency in early visual cortical areas. We re-evaluate this hypothesis by using natural and modified natural images to uncover the causal effects of luminance-contrast to human overt visual attention and: (i) we confirm that when viewing natural images, contrasts are elevated at fixation points. This, however, only holds for low spatial frequencies and in a limited temporal window after stimulus onset; (ii) however, despite this correlation between overt attention and luminance contrast, moderate modifications of contrast in natural images do not measurably affect the selection of fixation points. Furthermore, strong local reductions of luminance-contrast do not repel but attract fixation; (iii) neither contrast nor contrast modification is correlated to fixation duration; and (iv), even the moderate contrast modifications used fall into the physiologically relevant range, and subjects are well able to detect them in a forced choice paradigm. In summary, no causal contribution of luminance-contrast to a saliency map of human overt attention is detectable. In conjunction with recent results on the relation of contrast sensitivity of neuronal activity to the level in the visual cortical hierarchy, the present study provides evidence that, for natural scenes, saliency is computed not early but late during processing. PMID- 12653986 TI - The integration of cortical and behavioural dynamics during initial learning of a motor task. AB - Here we test the hypothesis that frequency and topographically specific changes in the strength of functional cortico-cortical coupling occur during the acquisition of a completely new task. To this end we studied the behavioural and cortical dynamics of a bimanual multifrequency coordination pattern during which one hand moved at twice the frequency of the other hand. This pattern represents a noninherent assignment and necessitates training before appropriate interlimb decoupling takes place. Results showed that acquisition of the multifrequency task was associated with an improved behavioural output that matched specific changes in the electroencephalogram dynamics. In particular, practice of the coordination pattern was accompanied by a decrease in coherence between the primary sensorimotor regions, and over the midline area in the alpha and beta bands, respectively, along with an increase in functional interhemispheric coupling between the prefrontal areas in the gamma band. These data suggest that the strength of cortico-cortical connectivity is adaptively modified across regions and across frequencies during early learning as the functional couplings are created and optimized for the purpose of movement execution. PMID- 12653987 TI - Functional properties and interaction of the anterior and posterior intraparietal areas in humans. AB - In the monkey the lateral bank of the anterior part of the intraparietal sulcus (area AIP), contains neurons that are involved in visually guided, object-related hand movements. It has also been shown that neurons in the caudal part of the intraparietal sulcus (area CIP) preferentially respond to 3D surface orientation. According to these results, it has been hypothesized that neurons in area CIP primarily encode the 3D features of an object and forwards this information to area AIP. AIP then utilizes this information for appropriate hand actions towards the object. Based on analogies to these primate studies, recent neuroimaging studies have suggested human homologues of areas AIP and CIP, however, the functional interaction between these areas remains unclear. Our event related fMRI study was designed to address specifically the question, how CIP and AIP interact in the process of adjustment of hand orientation towards objects. Volunteers were asked to perform three tasks: discrimination of surface orientation, imaging of visually guided hand movements and execution of visually guided hand movements. Our data show that the human AIP was activated both during discrimination of surface orientation and during the subsequent spatial adjustment of the thumb and index finger position towards the surface orientation. In contrast, human CIP was activated by the surface orientation but not by spatial adjustment of finger position. These data clearly indicate that the function of human CIP is more involved in coding 3D features of the objects, whereas human AIP is more involved in visually guided hand movements, similar to its role in the monkey. PMID- 12653988 TI - Developmental changes in calbindin-D28k and calretinin expression in the mouse suprachiasmatic nucleus. AB - The hypothalamic suprachiasmatic nucleus, the primary circadian pacemaker in mammals, and the retinohypothalamic tract, the retinal afferent fibres to the suprachiasmatic nucleus, both mature during early postnatal life. The establishment of circadian rhythms is thought to depend on input from the retina, but the mechanism remains unknown. Here we examined developmental changes in the expression of the Ca2+-binding proteins calbindin-D28k and calretinin in the mouse hypothalamus. Robust calbindin-D28k immunoreactivity was observed in the dorsomedial suprachiasmatic nucleus and the supraoptic nucleus in neonatal mice (postnatal day 3). The calbindin-D28k immunoreactivity decreased significantly in the suprachiasmatic nucleus but not in the supraoptic nucleus during postnatal days 9-15, when retinohypothalamic tract projections to the suprachiasmatic nucleus are completed. Calretinin immunoreactivity was low in the neonatal suprachiasmatic nucleus and increased with development in the ventrolateral suprachiasmatic nucleus, in parallel with the developmental reduction of calbindin-D28k immunoreactivity observed in the dorsomedial suprachiasmatic nucleus. Developmentally stable calretinin immunoreactivity was also observed in retinohypothalamic tract fibres. Organotypic slice cultures of the suprachiasmatic nucleus were prepared from postnatal day 3 mice to examine the effect of the absence of retinohypothalamic tract inputs on developmental changes in calbindin-D28k and calretinin expression. After 12 days in vitro, the cultured suprachiasmatic nucleus slices exhibited dense calbindin-D28k immunoreactivity similar to neonatal mice, and calretinin immunoreactivity in the ventrolateral suprachiasmatic nucleus similar to young adult mice. These results demonstrate a developmental reduction in calbindin-D28k expression that paralleled retinohypothalamic tract formation and a developmental increase in calretinin expression that is independent of retinohypothalamic tract connections to suprachiasmatic nucleus neurons. PMID- 12653989 TI - Working memory controls involuntary attention switching: evidence from an auditory distraction paradigm. AB - One function of working memory is to protect current mental processes against interference. In contrast, to be able to react flexibly on unpredictable environmental changes working memory should not totally be encapsulated from processing task unrelated information; that is, it should remain distractible. By manipulating the task load of the primary task in an auditory distraction paradigm we investigated how these opposing functions are coordinated by working memory. The behavioural results show that distraction effects were still present but reduced markedly with higher task demands. This suggests that working memory exerts some control over involuntary attention. In addition, event-related brain potentials related to the different processing stages reveal that the preattentive change detection system underlying distraction was not modulated by task demand whereas distraction per se was. The present data suggest that working memory is able to coordinate the maintenance of distractibility and the focus on the task at hand. PMID- 12653990 TI - The essential role of Broca's area in imitation. AB - The posterior sector of Broca's area (Brodmann area 44), a brain region critical for language, may have evolved from neurons active during observation and execution of manual movements. Imaging studies showing increased Broca's activity during execution, imagination, imitation and observation of hand movements support this hypothesis. Increased Broca's activity in motor task, however, may simply be due to inner speech. To test whether Broca's area is essential to imitation, we used repetitive transcranial magnetic stimulation (rTMS), which is known to transiently disrupt functions in stimulated areas. Subjects imitated finger key presses (imitation) or executed finger key presses in response to spatial cues (control task). While performing the tasks, subjects received rTMS over the left and right pars opercularis of the inferior frontal gyrus (where Brodmann area 44 is probabilistically located) and over the occipital cortex. There was significant impairment in imitation, but not in the control task, during rTMS over left and right pars opercularis compared to rTMS over the occipital cortex. This suggests that Broca's area is a premotor region essential to finger movement imitation. PMID- 12653991 TI - Enhancing cognitive performance with repetitive transcranial magnetic stimulation at human individual alpha frequency. AB - We applied rapid-rate repetitive transcranial magnetic stimulation (rTMS) at individual alpha frequency (IAF) to improve cognitive performance by influencing the dynamics of alpha desynchronization. Previous research indicates that a large upper alpha power in a reference interval preceding a task is related to both large suppression of upper alpha power during the task and good performance. Here, we tested the hypothesis that rTMS at individual upper alpha frequency (IAF + 1 Hz) can enhance alpha power in the reference interval, and can thus improve task performance. Repetitive TMS was delivered to the mesial frontal (Fz) and right parietal (P6) cortex, and as sham condition with 90 degrees-tilted coil (P6 position). The behavioural effect was assessed in a mental rotation task. Further control conditions were rTMS at a lower IAF (IAF - 3 Hz) and at 20 Hz. The results indicate that rTMS at IAF + 1 Hz can enhance task performance and, concomitantly, the extent of task-related alpha desynchronization. This provides further evidence for the functional relevance of oscillatory neuronal activity in the alpha band for the implementation of cognitive performance. PMID- 12653992 TI - Surface polysaccharide involvement in establishing the rhizobium-legume symbiosis. AB - When the rhizosphere is nitrogen-starved, legumes and rhizobia (soil bacteria) enter into a symbiosis that enables the fixation of atmospheric dinitrogen. This implies a complex chemical dialogue between partners and drastic changes on both plant roots and bacteria. Several recent works pointed out the importance of rhizobial surface polysaccharides in the establishing of the highly specific symbiosis between symbionts. Exopolysaccharides appear to be essential for the early infection process. Lipopolysaccharides exhibit specific roles in the later stages of the nodulation processes such as the penetration of the infection thread into the cortical cells or the setting up of the nitrogen-fixing phenotype. More generally, even if active at different steps of the establishing of the symbiosis, all the polysaccharide classes seem to be involved in complex processes of plant defense inhibition that allow plant root invasion. Their chemistry is important for structural recognition as well as for physico-chemical properties. PMID- 12653994 TI - In situ proton NMR analysis of alpha-alkynoate biotransformations. From 'invisible' substrates to detectable metabolites. AB - Only 2% of the known natural products with acetylenic bonds are alpha-alkynoates. Their polarized, conjugated triple bond is an optimal target for an enzymic hydration. Therefore they are good substrates for the enzymes involved in metabolism of acetylenic compounds, resulting in products that are suitable for bacterial growth. We isolated a Pseudomonas putida strain growing on 2 butynedioate as well as on propynoate, and determined the metabolic pathways of these two alpha-alkynoates. The triple bonds in both compounds were initially hydrated and 2-ketobutandioate as well as 3-ketopropanoate were formed. These two beta-keto acids were decarboxylated resulting in pyruvate and acetaldehyde, respectively. Pyruvate was further hydrolysed mainly to acetate and formate, whereas minor amounts were reduced to lactate. In the other biotransformation, acetaldehyde was oxidized to acetate accompanied by the reduction of 3 ketopropanoate to 3-hydroxypropanoate. Analyses of these metabolic processes were performed by in situ 1H-NMR spectroscopy in 1H2O, although the substrates, propynoate and 2-butynedioate, carried only one or even no detectable protons, respectively. However, while protons from the solvent are incorporated in the course of the pathway, the metabolites can be detected and identified. Therefore a detailed determination of the metabolic process is possible. PMID- 12653993 TI - 'Antifreeze' glycoproteins from polar fish. AB - Antifreeze glycoproteins (AFGPs) constitute the major fraction of protein in the blood serum of Antarctic notothenioids and Arctic cod. Each AFGP consists of a varying number of repeating units of (Ala-Ala-Thr)n, with minor sequence variations, and the disaccharide beta-D-galactosyl-(1-->3)-alpha-N-acetyl-D galactosamine joined as a glycoside to the hydroxyl oxygen of the Thr residues. These compounds allow the fish to survive in subzero ice-laden polar oceans by kinetically depressing the temperature at which ice grows in a noncolligative manner. In contrast to the more widely studied antifreeze proteins, little is known about the mechanism of ice growth inhibition by AFGPs, and there is no definitive model that explains their properties. This review summarizes the structural and physical properties of AFGPs and advances in the last decade that now provide opportunities for further research in this field. High field NMR spectroscopy and molecular dynamics studies have shown that AFGPs are largely unstructured in aqueous solution. While standard carbohydrate degradation studies confirm the requirement of some of the sugar hydroxyls for antifreeze activity, the importance of following structural elements has not been established: (a) the number of hydroxyls required, (b) the stereochemistry of the sugar hydroxyls (i.e. the requirement of galactose as the sugar), (c) the acetamido group on the first galactose sugar, (d) the stereochemistry of the beta-glycosidic linkage between the two sugars and the alpha-glycosidic linkage to Thr, (e) the requirement of a disaccharide for activity, and (f) the Ala and Thr residues in the polypeptide backbone. The recent successful synthesis of small AFGPs using solution methods and solid-phase chemistry provides the opportunity to perform key structure-activity studies that would clarify the important residues and functional groups required for activity. Genetic studies have shown that the AFGPs present in the two geographically and phylogenetically distinct Antarctic notothenioids and Arctic cod have evolved independently, in a rare example of convergent molecular evolution. The AFGPs exhibit concentration dependent thermal hysteresis with maximum hysteresis (1.2 degrees C at 40 mg x mL-1) observed with the higher molecular mass glycoproteins. The ability to modify the rate and shape of crystal growth and protect cellular membranes during lipid-phase transitions have resulted in identification of a number of potential applications of AFGPs as food additives, and in the cryopreservation and hypothermal storage of cells and tissues. PMID- 12653995 TI - Three-dimensional structures of thermophilic beta-1,4-xylanases from Chaetomium thermophilum and Nonomuraea flexuosa. Comparison of twelve xylanases in relation to their thermal stability. AB - The crystal structures of thermophilic xylanases from Chaetomium thermophilum and Nonomuraea flexuosa were determined at 1.75 and 2.1 A resolution, respectively. Both enzymes have the overall fold typical to family 11 xylanases with two highly twisted beta-sheets forming a large cleft. The comparison of 12 crystal structures of family 11 xylanases from both mesophilic and thermophilic organisms showed that the structures of different xylanases are very similar. The sequence identity differences correlated well with the structural differences. Several minor modifications appeared to be responsible for the increased thermal stability of family 11 xylanases: (a) higher Thr : Ser ratio (b) increased number of charged residues, especially Arg, resulting in enhanced polar interactions, and (c) improved stabilization of secondary structures involved the higher number of residues in the beta-strands and stabilization of the alpha-helix region. Some members of family 11 xylanases have a unique strategy to improve their stability, such as a higher number of ion pairs or aromatic residues on protein surface, a more compact structure, a tighter packing, and insertions at some regions resulting in enhanced interactions. PMID- 12653996 TI - Steady-state and time-resolved fluorescence studies of conformational changes induced by cyclic AMP and DNA binding to cyclic AMP receptor protein from Escherichia coli. AB - cAMP receptor protein (CRP), allosterically activated by cAMP, regulates the expression of several genes in Escherichia coli. As binding of cAMP leads to undefined conformational changes in CRP, we performed a steady-state and time resolved fluorescence study to show how the binding of the ligand influences the structure and dynamics of the protein. We used CRP mutants containing a single tryptophan residue at position 85 or 13, and fluorescently labeled with 1,5-I AEDANS attached to Cys178. Binding of cAMP in the CRP-(cAMP)2 complex leads to changes in the Trp13 microenvironment, whereas its binding in the CRP-(cAMP)4 complex alters the surroundings of Trp85. Time-resolved anisotropy measurements indicated that cAMP binding in the CRP-(cAMP)2 complex led to a substantial increase in the rotational mobility of the Trp13 residue. Measurement of fluorescence energy transfer (FRET) between labeled Cys178 and Trp85 showed that the binding of cAMP in the CRP-(cAMP)2 complex caused a substantial increase in FRET efficiency. This indicates a decrease in the distance between the two domains of the protein from 26.6 A in apo-CRP to 18.7 A in the CRP-(cAMP)2 complex. The binding of cAMP in the CRP-(cAMP)4 complex resulted in only a very small increase in FRET efficiency. The average distance between the two domains in CRP-DNA complexes, possessing lac, gal or ICAP sequences, shows an increase, as evidenced by the increase in the average distance between Cys178 and Trp85 to approximately 20 A. The spectral changes observed provide new structural information about the cAMP-induced allosteric activation of the protein. PMID- 12653997 TI - The swinging movement of the distal histidine residue and the autoxidation reaction for midge larval hemoglobins. AB - Some insects have a globin exclusively in their fast-growing larval stage. This is the case in the 4th-instar larva of Tokunagayusurika akamusi, a common midge found in Japan. In the polymorphic hemoglobin comprised of 11 separable components, hemoglobin VII (Ta-VII Hb) was of particular interest. When its ferric met-form was exposed to pH 5.0 from 7.2, the distal histidine was found to swing away from the E7 position. As a result, the iron(III) was converted from a hexacoordinate to a pentacoordinate form by a concomitant loss of the axial water ligand. The corresponding spectral changes in the Soret band were therefore followed by stopped-flow and rapid-scan techniques, and the observed first-order rate constants of k(out) = 25 s(-1) and kin = 128 s(-1) were obtained for the outward and inward movements, respectively, of the distal histidine residue in 0.1 m buffer at 25 degrees C. For O2 affinity, Ta-VII Hb showed a value of P50 = 1.7 Torr at pH 7.4, accompanied with a remarkable Bohr effect (deltaH+ = -0.58) almost equal to that of mammalian hemoglobins. We have also investigated the stability property of Ta-VII HbO2 in terms of the autoxidation rate over a wide range of pH from 4 to 11. The resulting pH-dependence curve was compared with those of another component Ta-V HbO2 and sperm whale MbO2, and described based on a nucleophilic displacement mechanism. In light of the O2 binding affinity, Bohr effect and considerable stability of the bound O2 against acidic autoxidation, we conclude that T. akamusi Hb VII can play an important role in O2 transport and storage as the major component in the larval hemolymph. PMID- 12653998 TI - Phenol hydroxylase from Acinetobacter radioresistens S13. Isolation and characterization of the regulatory component. AB - This paper reports the isolation and characterization of the regulatory moiety of the multicomponent enzyme phenol hydroxylase from Acinetobacter radioresistens S13 grown on phenol as the only carbon and energy source. The whole enzyme comprises an oxygenase moiety (PHO), a reductase moiety (PHR) and a regulatory moiety (PHI). PHR contains one FAD and one iron-sulfur cluster, whose function is electron transfer from NADH to the dinuclear iron centre of the oxygenase. PHI is required for catalysis of the conversion of phenol to catechol in vitro, but is not required for PHR activity towards alternative electron acceptors such as cytochrome c and Nitro Blue Tetrazolium. The molecular mass of PHI was determined to be 10 kDa by SDS/PAGE, 8.8 kDa by MALDI-TOF spectrometry and 18 kDa by gel permeation. This finding suggests that the protein in its native state is a homodimer. The isoelectric point is 4.1. PHI does not contain any redox cofactor and does not bind ANS, a fluorescent probe for hydrophobic sites. The N-terminal sequence is similar to those of the regulatory proteins of phenol hydroxylase from A. calcoaceticus and Pseudomonas CF 600. In the reconstituted system, optimal reaction rate was achieved when the stoichiometry of the components was 2 PHR monomers: 1 PHI dimer: 1 PHO (alphabetagamma) dimer. PHI interacts specifically with PHR, promoting the enhancement of FAD fluorescence emission. This signal is diagnostic of a conformational change of PHR that might result in a better alignment with respect to PHO. PMID- 12653999 TI - Association of feather colour with constitutively active melanocortin 1 receptors in chicken. AB - Seven alleles of the chicken melanocortin (MC) 1 receptor were cloned into expression vectors, expressed in mammalian cells and pharmacologically characterized. Four of the clones e(+R), e(+B&D), e(wh)/e(y), E(Rfayoumi) gave receptors to which melanocortin stimulating hormone (alpha-MSH) and NDP-MSH bound with similar IC50 values and responded to alpha-MSH by increasing intracellular cAMP levels in a dose-dependent manner. Three of the cMC1 receptors; e(b), E and E(R), did not show any specific binding to the radioligand, but were found to be constitutively active in the cAMP assay. The E and E(R) alleles are associated with black feather colour in chicken while the eb allele gives rise to brownish pigmentation. The three constitutively active receptors share a mutation of Glu to Lys in position 92. This mutation was previously found in darkly pigmented sombre mice, but constitutively active MC receptors have not previously been shown in any nonmammalian species. We also inserted the Glu to Lys mutation in the human MC1 and MC4 receptors. In contrast with the chicken clones, the hMC1 E94K receptor bound to the ligand, but was still constitutively active independently of ligand concentration. The hMC4-E100K receptor did not bind to the MSH ligand and was not constitutively active. The results indicate that the structural requirements that allow the receptor to adapt an active conformation without binding to a ligand, as a consequence of this E/K mutation, are not conserved within the MC receptors. The results are discussed in relationship to feather colour in chicken, molecular receptor structures and evolution. We suggest that properties for the 'E92K switch' mechanism may have evolved in an ancestor common to chicken and mammals and were maintained over long time periods through evolutionary pressure, probably on closely linked structural features. PMID- 12654001 TI - Preventing nondesired RNA-primed RNA extension catalyzed by T7 RNA polymerase. AB - The transcription patterns of 64 linear double stranded DNA templates obtained with T7 RNA polymerase were investigated. These templates consisted of 17 nucleotide-long sequences under the control of the minimal bacteriophage T7 promoter and represented all possible combinations of nucleotides at positions +8, +10 and +11. Two clearly distinct types of template were identified, which produced the range of transcription patterns observed: (a) those that yielded 17 nucleotide-long RNA as the only detectable run-off product (only 15% of the total), and (b) templates that in addition to the expected full-length RNA, produced other products longer than 17 nucleotides. Self-complementarity analysis of the expected run-off transcripts showed that those obtained from the first type of template were able to form stable intermolecular duplexes with non-base paired 3'-ends. However, the second type of template yielded RNAs able to generate energetically favorable intermolecular duplexes with 3'-end complementarity, therefore yielding an RNA-primed RNA-template. The gel-purified 17-nucleotide-long RNAs transcribed from the latter yielded longer products when incubated under in vitro transcription conditions in the absence of a DNA template. No extension was observed when assaying the 17-nucleotide RNA products resulting from the first type of template. We observed that just a single nucleotide change within the DNA template could convert the RNA product from an RNA-primed template into a nonextendible dimer thus leading to a drastic switch of the 17-nucleotide product yield from less than 10% to 100%. Further, two type B DNA templates were extended by two nucleotides at the 3'-end, to produce RNA transcripts theoretically unable to form 3'-end base-paired duplexes. The full length products of these modified DNA templates were found to be nonextendible by T7 RNA polymerase under the standard in vitro transcription conditions. PMID- 12654000 TI - Connection of transport and sensing by UhpC, the sensor for external glucose-6 phosphate in Escherichia coli. AB - UhpC is a membrane-bound sensor protein in Escherichia coli required for recognizing external glucose-6-phosphate (Glc6P) and induction of the transport protein UhpT. Recently, it was shown that UhpC is also able to transport Glc6P. In this study we investigated whether these transport and sensing activities are obligatorily coupled in UhpC. We expressed a His-UhpC protein in a UhpC-deficient E. coli strain and verified that this construct does not alter the basic biochemical properties of the Glc6P sensor system. The effects of arginine replacements, mutations of the central loop, and introduction of a salt bridge in UhpC on transport and sensing were compared. The exchanges R46C, R266C and R149C moderately affected transport by UhpC but strongly decreased the sensing ability. This suggested that the affinity for Glc6P as a transported substrate is uncoupled in UhpC from its affinity for Glc6P as an inducer. Four of the 11 arginine mutants showed a constitutive phenotype but had near wild-type transport activity suggesting that Glc6P can be transported by a molecule locked in the inducing conformation. Introduction of an intrahelical salt bridge increased the transport activity of UhpC but abolished sensing. Three conserved residues from the central loop were mutated and although none of these showed transport, one exhibited increased affinity for sensing. Taken together, these data show that transport by UhpC is not required for sensing, that conserved arginine residues are important for sensing and not for transport, and that residues located in the central hydrophilic loop are critical for transport and for sensing. PMID- 12654002 TI - Mechanisms of accumulation of arachidonate in phosphatidylinositol in yellowtail. A comparative study of acylation systems of phospholipids in rat and the fish species Seriola quinqueradiata. AB - It is known that phosphatidylinositol (PtdIns) contains abundant arachidonate and is composed mainly of 1-stearoyl-2-arachidonoyl species in mammals. We investigated if this characteristic of PtdIns applies to the PtdIns from yellowtail (Seriola quinqueradiata), a marine fish. In common with phosphatidylcholine (PtdCho), phosphatidylethanolamine (PtdEtn) and phosphatidylserine (PtdSer) from brain, heart, liver, spleen, kidney and ovary, the predominant polyunsaturated fatty acid was docosahexaenoic acid, and levels of arachidonic acid were less than 4.5% (PtdCho), 7.5% (PtdEtn) and 3.0% (PtdSer) in these tissues. In striking contrast, arachidonic acid made up 17.6%, 31.8%, 27.8%, 26.1%, 25.4% and 33.5% of the fatty acid composition of PtdIns from brain, heart, liver, spleen, kidney and ovary, respectively. The most abundant molecular species of PtdIns in all these tissues was 1-stearoyl-2-arachidonoyl. Assay of acyltransferase in liver microsomes of yellowtail showed that arachidonic acid was incorporated into PtdIns more effectively than docosahexaenoic acid and that the latter inhibited incorporation of arachidonic acid into PtdCho without inhibiting the utilization of arachidonic acid for PtdIns. This effect of docosahexaenoic acid was not observed in similar experiments using rat liver microsomes and is thought to contribute to the exclusive utilization of arachidonic acid for acylation to PtdIns in yellowtail. Inositolphospholipids and their hydrolysates are known to act as signaling molecules in cells. The conserved hydrophobic structure of PtdIns (the 1-stearoyl-2-arachidonoyl moiety) may have physiological significance not only in mammals but also in fish. PMID- 12654003 TI - Kinetic mechanisms of glycine oxidase from Bacillus subtilis. AB - The kinetic properties of glycine oxidase from Bacillus subtilis were investigated using glycine, sarcosine, and d-proline as substrate. The turnover numbers at saturating substrate and oxygen concentrations were 4.0 s(-1), 4.2 s( 1), and 3.5 s(-1), respectively, with glycine, sarcosine, and D-proline as substrate. Glycine oxidase was converted to a two-electron reduced form upon anaerobic reduction with the individual substrates and its reductive half reaction was demonstrated to be reversible. The rates of flavin reduction extrapolated to saturating substrate concentration, and under anaerobic conditions, were 166 s(-1), 170 s(-1), and 26 s(-1), respectively, with glycine, sarcosine, and D-proline as substrate. The rate of reoxidation of reduced glycine oxidase with oxygen in the absence of product (extrapolated rate approximately 3 x 10(4) M(-1) x s(-1)) was too slow to account for catalysis and thus reoxidation started from the reduced enzyme:imino acid complex. The kinetic data are compatible with a ternary complex sequential mechanism in which the rate of product dissociation from the reoxidized enzyme form represents the rate-limiting step. Although glycine oxidase and D-amino acid oxidase differ in substrate specificity and amino acid sequence, the kinetic mechanism of glycine oxidase is similar to that determined for mammalian D-amino acid oxidase on neutral D-amino acids, further supporting a close similarity between these two amine oxidases. PMID- 12654004 TI - The nucleotide-binding domains of P-glycoprotein. Functional symmetry in the isolated domain demonstrated by N-ethylmaleimide labelling. AB - The two nucleotide-binding domains (NBDs) of a number of ATP-binding cassette (ABC) transporters have been shown to be functionally dissimilar, playing different roles in the transport process. A high degree of co-operativity has been determined for the NBDs of the human multidrug transporter, P-glycoprotein. However, the issue of functional symmetry in P-glycoprotein remains contentious. To address this, the NBDs of P-glycoprotein were expressed and purified to 95% homogeneity, as fusions to maltose-binding protein. The NBDs were engineered to contain a single cysteine residue in the Walker-A homology motif. Reactivity of this cysteine residue was demonstrated by specific, time-dependent, covalent labelling with N-ethylmaleimide. No differences in the rates of labelling of the two NBDs were observed. The relative affinity of binding to each NBD was determined for a number of nucleotides by measuring their ability to effect a reduction in N-ethylmaleimide labelling. In general, nucleotides bound identically to the two NBDs, suggesting that there is little asymmetry in the initial step of the transport cycle, namely the recognition and binding of nucleotide. Any observed functional asymmetry in the intact transporter presumably reflects different rates of hydrolysis at the two NBDs or interdomain communications. PMID- 12654006 TI - Critical role of the plasma membrane for expression of mammalian mitochondrial side chain cleavage activity in yeast. AB - Engineered yeast cells efficiently convert ergosta-5-eneol to pregnenolone and progesterone provided that endogenous pregnenolone acetylase activity is disrupted and that heterologous sterol delta7-reductase, cytochrome P450 side chain cleavage (CYP11A1) and 3beta hydroxysteroid dehydrogenase/isomerase (3beta HSD) activities are present. CYP11A1 activity requires the expression of the mammalian NADPH-adrenodoxin reductase (Adrp) and adrenodoxin (Adxp) proteins as electron carriers. Several parameters modulate this artificial metabolic pathway: the effects of steroid products; the availability and delivery of the ergosta-5 eneol substrate to cytochrome P450; electron flux and protein localization. CYP11A1, Adxp and Adrp are usually located in contact with inner mitochondrial membranes and are directed to the outside of the mitochondria by the removal of their respective mitochondrial targeting sequences. CYP11A1 then localizes to the plasma membrane but Adrp and Adxp are detected in the endoplasmic reticulum and cytosol as expected. The electron transfer chain that involves several subcellular compartments may control side chain cleavage activity in yeast. Interestingly, Tgl1p, a potential ester hydrolase, was found to enhance steroid productivity, probably through both the availability and/or the trafficking of the CYP11A1 substrate. Thus, the observation that the highest cellular levels of free ergosta-5-eneol are found in the plasma membrane suggests that the substrate is freely available for pregnenolone synthesis. PMID- 12654005 TI - In vivo studies of altered expression patterns of p53 and proliferative control genes in chronic vitamin A deficiency and hypervitaminosis. AB - Several clinical trials have revealed that individuals who were given beta carotene and vitamin A did not have a reduced risk of cancer compared to those given placebo; rather, vitamin A could actually have caused an adverse effect in the lungs of smokers [Omenn, G.S., Goodman, G.E., Thornquist, M.D., Balmes, J., Cullen, M.R., Glass, A., Keogh, J.P., Meyskens, F.L., Valanis, B., Williams, J.H., Barnhart, S. & Hammar, S. N. Engl. J. Med (1996) 334, 1150-1155; Hennekens, C.H., Buring, J.E., Manson, J.E., Stampfer, M., Rosner, B., Cook, N.R., Belanger, C., LaMotte, F., Gaziano, J.M., Ridker, P.M., Willet, W. & Peto, R. (1996) N. Engl. J. Med. 334, 1145-1149]. Using differential display techniques, an initial survey using rats showed that liver RNA expression of c-H-Ras was decreased and p53 increased in rats with chronic vitamin A deficiency. These findings prompted us to evaluate the expression of c-Jun, p53 and p21WAF1/CIF1 (by RT-PCR) in liver and lung of rats. This study showed that c-Jun levels were lower and that p53 and p21WAF1/CIF1 levels were higher in chronic vitamin A deficiency. Vitamin A supplementation increased expression of c-Jun, while decreasing the expression of p53 and p21WAF1/CIF1. Western-blot analysis demonstrated that c-Jun and p53 showed a similar pattern to that found in the RT-PCR analyses. Binding of retinoic acid receptors (RAR) to the c-Jun promoter was decreased in chronic vitamin A deficiency when compared to control hepatocytes, but contrasting results were found with acute vitamin A supplementated cells. DNA fragmentation and cytochrome c release from mitochondria were analyzed and no changes were found. In lung, an increase in the expression of c-Jun produced a significant increase in cyclin D1 expression. These results may explain, at least in part, the conflicting results found in patients supplemented with vitamin A and illustrate that the changes are not restricted to lung. Furthermore, these results suggest that pharmacological vitamin A supplementation may increase the risk of adverse effects including the risk of oncogenesis. PMID- 12654007 TI - Functional fine-mapping and molecular modeling of a conserved loop epitope of the measles virus hemagglutinin protein. AB - Neutralizing and protective monoclonal antibodies (mAbs) were used to fine-map the highly conserved hemagglutinin noose epitope (H379-410, HNE) of the measles virus. Short peptides mimicking this epitope were previously shown to induce virus-neutralizing antibodies [El Kasmi et al. (2000) J. Gen. Virol.81, 729-735]. The epitope contains three cysteine residues, two of which (Cys386 and Cys394) form a disulfide bridge critical for antibody binding. Substitution and truncation analogues revealed four residues critical for binding (Lys387, Gly388, Gln391 and Glu395) and suggested the binding motif X7C[KR]GX[AINQ]QX2CEX5 for three distinct protective mAbs. This motif was found in more than 90% of the wild type viruses. An independent molecular model of the core epitope predicted an amphiphilic loop displaying a remarkably stable and rigid loop conformation. The three hydrophilic contact residues Lys387, Gln391 and Glu395 pointed on the virus towards the solvent-exposed side of the planar loop and the permissive hydrophobic residues Ile390, Ala392 and Leu393 towards the solvent-hidden side of the loop, precluding antibody binding. The high affinity (Kd = 7.60 nm) of the mAb BH216 for the peptide suggests a high structural resemblance of the peptide with the natural epitope and indicates that most interactions with the protein are also contributed by the peptide. Improved peptides designed on the basis of these findings induced sera that crossreacted with the native measles virus hemagglutinin protein, providing important information about a lead structure for the design of more stable antigens of a synthetic or recombinant subunit vaccine. PMID- 12654008 TI - The N-terminal cysteine pair of yeast sulfhydryl oxidase Erv1p is essential for in vivo activity and interacts with the primary redox centre. AB - Yeast Erv1p is a ubiquitous FAD-dependent sulfhydryl oxidase, located in the intermembrane space of mitochondria. The dimeric enzyme is essential for survival of the cell. Besides the redox-active CXXC motif close to the FAD, Erv1p harbours two additional cysteine pairs. Site-directed mutagenesis has identified all three cysteine pairs as essential for normal function. The C-terminal cysteine pair is of structural importance as it contributes to the correct arrangement of the FAD binding fold. Variations in dimer formation and unique colour changes of mutant proteins argue in favour of an interaction between the N-terminal cysteine pair with the redox centre of the partner monomer. PMID- 12654009 TI - Localization of fluorescence-labeled poly(malic acid) to the nuclei of the plasmodium of Physarum polycephalum. AB - The nuclei in the plasmodium of Physarum polycephalum, as of other myxomycetes, contain high amounts of polymalate, which has been proposed to function as a scaffold for the carriage and storage of several DNA-binding proteins [Angerer, B. and Holler, E. (1995) Biochemistry 34, 14741-14751]. By delivering fluorescence-labeled polymalate into a growing plasmodium by injection, we observed microscopic staining of nuclei in agreement with the proposed function. The fluorescence intensity was highest during the reconstruction phase of the nuclei. To examine whether the delivery was under the control of polymalatase or related proteins [Karl, M. & Holler, E. (1998) Eur. J. Biochem.251, 405-412], the cellular distribution of these proteins was also examined by staining with antibodies against polymalatase. Double-stained plasmodia revealed a fluorescent halo around each fluorescent nucleus during the reconsititution. Fluorescent nuclei were not observed when the hydroxyl terminus of polymalate, known to be essential for the binding of polymalatase, was blocked by labeling with fluorescein-5-isothiocyanate. By immune precipitation, it was shown that polymalate and polymalatase or related proteins were in the precipitate. It is concluded that polymalate is delivered to the surface of nuclei in the complex with polymalatase or related proteins. The complex dissociates, and polymalate translocates into the nucleus, while polymalatase or related proteins remain at the surface. PMID- 12654010 TI - In vitro analysis of the relationship between endonuclease and maturase activities in the bi-functional group I intron-encoded protein, I-AniI. AB - The AnCOB group I intron from Aspergillus nidulans encodes a homing DNA endonuclease called I-AniI which also functions as a maturase, assisting in AnCOB intron RNA splicing. In this investigation we biochemically characterized the endonuclease activity of I-AniI in vitro and utilized competition assays to probe the relationship between the RNA- and DNA-binding sites. Despite functioning as an RNA maturase, I-AniI still retains several characteristic properties of homing endonucleases including relaxed substrate specificity, DNA cleavage product retention and instability in the reaction buffer, which suggest that the protein has not undergone dramatic structural adaptations to function as an RNA-binding protein. Nitrocellulose filter binding and kinetic burst assays showed that both nucleic acids bind I-AniI with the same 1 : 1 stoichiometry. Furthermore, in vitro competition activity assays revealed that the RNA substrate, when prebound to I-AniI, stoichiometrically inhibits DNA cleavage activity, yet in reciprocal experiments, saturating amounts of prebound DNA substrate fails to inhibit RNA splicing activity. The data suggest therefore that both nucleic acids do not bind the same single binding site, rather that I-AniI appears to contain two binding sites. PMID- 12654011 TI - Oxygen control of nif gene expression in Klebsiella pneumoniae depends on NifL reduction at the cytoplasmic membrane by electrons derived from the reduced quinone pool. AB - In Klebsiella pneumoniae, the flavoprotein, NifL regulates NifA mediated transcriptional activation of the N2-fixation (nif) genes in response to molecular O2 and ammonium. We investigated the influence of membrane-bound oxidoreductases on nif-regulation by biochemical analysis of purified NifL and by monitoring NifA-mediated expression of nifH'-'lacZ reporter fusions in different mutant backgrounds. NifL-bound FAD-cofactor was reduced by NADH only in the presence of a redox-mediator or inside-out vesicles derived from anaerobically grown K. pneumoniae cells, indicating that in vivo NifL is reduced by electrons derived from membrane-bound oxidoreductases of the anaerobic respiratory chain. This mechanism is further supported by three lines of evidence: First, K. pneumoniae strains carrying null mutations of fdnG or nuoCD showed significantly reduced nif-induction under derepressing conditions, indicating that NifL inhibition of NifA was not relieved in the absence of formate dehydrogenase-N or NADH:ubiquinone oxidoreductase. The same effect was observed in a heterologous Escherichia coli system carrying a ndh null allele (coding for NADH dehydrogenaseII). Second, studying nif-induction in K. pneumoniae revealed that during anaerobic growth in glycerol, under nitrogen-limitation, the presence of the terminal electron acceptor nitrate resulted in a significant decrease of nif induction. The final line of evidence is that reduced quinone derivatives, dimethylnaphthoquinol and menadiol, are able to transfer electrons to the FAD moiety of purified NifL. On the basis of these data, we postulate that under anaerobic and nitrogen-limited conditions, NifL inhibition of NifA activity is relieved by reduction of the FAD-cofactor by electrons derived from the reduced quinone pool, generated by anaerobic respiration, that favours membrane association of NifL. We further hypothesize that the quinol/quinone ratio is important for providing the signal to NifL. PMID- 12654012 TI - Functional expression of the quinoline 2-oxidoreductase genes (qorMSL) in Pseudomonas putida KT2440 pUF1 and in P. putida 86-1 deltaqor pUF1 and analysis of the Qor proteins. AB - The availability of a system for the functional expression of genes coding for molybdenum hydroxylases is a prerequisite for the construction of enzyme variants by mutagenesis. For the expression cloning of quinoline 2-oxidoreductase (Qor) from Pseudomonas putida 86--that contains the molybdopterin cytosine dinucleotide molybdenum cofactor (Mo-MCD), two distinct [2Fe-2S] clusters and FAD--the qorMSL genes were inserted into the broad host range vector, pJB653, generating pUF1. P. putida KT2440 and P. putida 86-1 deltaqor were used as recipients for pUF1. Whereas Qor from the wild-type strain showed a specific activity of 19-23 U x mg( 1), the specific activity of Qor purified from P. putida KT2440 pUF1 was only 0.8 2.5 U x mg(-1), and its apparent k(cat) (quinoline) was about ninefold lower than that of wild-type Qor. The apparent Km values for quinoline were similar for both proteins. UV/visible and EPR spectroscopy indicated the presence of the full set of [2Fe-2S] clusters and FAD in Qor from P. putida KT2440 pUF1, however, the very low intensity of the Mo(V)-rapid signal, that occurs in the presence of quinoline, as well as metal analysis indicated a deficiency of the molybdenum center. In contrast, the metal content, and the spectroscopic and catalytic properties of Qor produced by P. putida 86-1 deltaqor pUF1 were essentially like those of wild-type Qor. Release of CMP upon acidic hydrolysis of the Qor proteins suggested the presence of the MCD form of the pyranopterin cofactor; the CMP contents of the three enzymes were similar. PMID- 12654013 TI - H2O2, but not menadione, provokes a decrease in the ATP and an increase in the inosine levels in Saccharomyces cerevisiae. An experimental and theoretical approach. AB - When Saccharomyces cerevisiae cells, grown in galactose, glucose or mannose, were treated with 1.5 mm hydrogen peroxide (H2O2) for 30 min, an important decrease in the ATP, and a less extensive decrease in the GTP, CTP, UTP and ADP-ribose levels was estimated. Concomitantly a net increase in the inosine levels was observed. Treatment with 83 mm menadione promoted the appearance of a compound similar to adenosine but no appreciable changes in the nucleotide content of yeast cells, grown either in glucose or galactose. Changes in the specific activities of the enzymes involved in the pathway from ATP to inosine, in yeast extracts from (un)treated cells, could not explain the effect of H2O2 on the levels of ATP and inosine. Application of a mathematical model of differential equations previously developed in this laboratory pointed to a potential inhibition of glycolysis as the main reason for that effect. This theoretical consideration was reinforced both by the lack of an appreciable effect of 1.5 mm (or even higher concentrations) H2O2 on yeast grown in the presence of ethanol or glycerol, and by the observed inhibition of the synthesis of ethanol promoted by H2O2. Normal values for the adenylic charge, ATP and inosine levels were reached at 5, 30 and 120 min, respectively, after removal of H2O2 from the culture medium. The strong decrease in the ATP level upon H2O2 treatment is an important factor to be considered for understanding the response of yeast, and probably other cell types, to oxidative stress. PMID- 12654014 TI - Deadenylation of interferon-beta mRNA is mediated by both the AU-rich element in the 3'-untranslated region and an instability sequence in the coding region. AB - Viral infection of fibroblastic and endothelial cells leads to the transient synthesis of interferon-beta (IFN-beta). The down-regulation of IFN-beta synthesis after infection results both from transcriptional repression of the IFN beta gene and rapid degradation of mRNA. As with many cytokine mRNAs, IFN-beta mRNA contains an AU-rich element (ARE) in its 3'-untranslated region (UTR). AREs are known to mediate mRNA deadenylation and destabilization. Depending on the class of ARE, deadenylation was shown to occur through synchronous or asynchronous mechanisms. In this study, we analysed IFN-beta mRNA deadenylation in natural conditions of IFN-beta synthesis, e.g., after viral infection. We show that human IFN-beta mRNA follows an asynchronous deadenylation pathway typical of a mRNA containing a class II ARE. A deletion analysis of the IFN-beta natural transcript demonstrates that poly(A) shortening can be mediated by the ARE but also by a 32 nucleotide-sequence located in the coding region, that was identified previously as an instability determinant. In fact, these elements are able to act independently as both of them have to be removed to abrogate mRNA deadenylation. Our data also indicate that deadenylation occurs independently of mRNA translation. Moreover, we show that deadenylation of IFN-beta mRNA is not under the control of viral infection as IFN-beta mRNA derived from a constitutively expressed gene cassette is deadenylated in absence of viral infection. Finally, an unidentified nuclear event appears to be a prerequisite for IFN-beta mRNA deadenylation as IFN-beta mRNA introduced directly into the cytoplasm does not undergo deadenylation. In conclusion, our study demonstrates that IFN-beta mRNA poly(A) shortening is under the control of two cis-acting elements recruiting a deadenylating machinery whose activity is independent of translation and viral infection but might require a nuclear event. PMID- 12654015 TI - Wound bed preparation: a systematic approach to wound management. AB - The healing process in acute wounds has been extensively studied and the knowledge derived from these studies has often been extrapolated to the care of chronic wounds, on the assumption that nonhealing chronic wounds were simply aberrations of the normal tissue repair process. However, this approach is less than satisfactory, as the chronic wound healing process differs in many important respects from that seen in acute wounds. In chronic wounds, the orderly sequence of events seen in acute wounds becomes disrupted or "stuck" at one or more of the different stages of wound healing. For the normal repair process to resume, the barrier to healing must be identified and removed through application of the correct techniques. It is important, therefore, to understand the molecular events that are involved in the wound healing process in order to select the most appropriate intervention. Wound bed preparation is the management of a wound in order to accelerate endogenous healing or to facilitate the effectiveness of other therapeutic measures. Experts in wound management consider that wound bed preparation is an important concept with significant potential as an educational tool in wound management. This article was developed after a meeting of wound healing experts in June 2002 and is intended to provide an overview of the current status, role, and key elements of wound bed preparation. Readers will be able to examine the following issues; the current status of wound bed preparation; an analysis of the acute and chronic wound environments; how wound healing can take place in these environments; the role of wound bed preparation in the clinic; the clinical and cellular components of the wound bed preparation concept; a detailed analysis of the components of wound bed preparation. PMID- 12654018 TI - In vitro development of marmoset monkey oocytes by pre-antral follicle culture. AB - A technique for in vitro maturation of oocytes from small ovarian follicles of marmoset monkeys (Callithrix jacchus) has been developed. We employed a two-step culture system for primary follicles (45-85 microm) and a one-step culture technique for secondary follicles (>85 microm). The two-step technique started with the culture of stromal tissue fragments for 2 days. Thereafter, mechanically isolated follicles were transferred to a culture system where they attached to the culture surface and grew for up to a further 12 days. Significant growth of the small follicles and their oocytes was only achieved with gonadotrophins in the medium. Oocytes with a mean diameter of 39 microm from follicles <85 microm reached a mean diameter of 90 microm by the end of the two-step culture. After in vitro maturation, 19% of oocytes from these follicles had progressed to the germinal vesicle breakdown (GVBD). Follicles between 85 and 170 microm in diameter were isolated from the stroma and placed directly in the culture. Oocytes from these follicles had a mean diameter of 64 microm. The maximum size the oocytes reached in culture was related to the age of the females (pre pubertal females: 102 +/- 1.3 microm; adults: 96 +/- 1.4 microm). Twenty-seven per cent of oocytes from pre-pubertal ovaries achieved GVBD and nearly two-thirds of these progressed to polar body stage. From adult ovaries, only 12% progressed to GVBD and one-third of these to polar body stage. It is possible to develop mature oocytes in vitro from marmoset secondary pre-antral follicles (>85 microm). From primary follicles, although near full size oocytes were developed, maturation capacity was incomplete. PMID- 12654017 TI - Advances in biotechnology: new tools in future pig production for agriculture and biomedicine. AB - Biotechnology in livestock comprises an arsenal of reproductive biotechniques and molecular genetics. While molecular genetics are poorly developed in swine, reproductive techniques are more advanced and applied under field conditions. This review describes three selected examples of our own research to illustrate the implication of biotechnology in future pig reproduction. Sperm sexing technology is now available and can be used to generate piglets of the desired sex by IVF and ICSI. First studies also indicate satisfactory success rates following intrauterine insemination with sexed spermatozoa. Cloning technique and production of transgenic pigs require information about the regulation and time course of gene expression during in vitro production and pre-implantation development. Information on gene expression is scare in porcine embryos. With the exception of transcripts for the oestrogen receptor gene, no mRNA's from the activated porcine genome have been identified. Recent development of cDNA arrays might help to identify a larger amount of genes in single embryos. Remarkable progress has been made in organ transplantation technology. As the demand for human organs is increasing rapidly, the pig might serve as donor of xenotranplants, provided the transmission of zoonoses from the donor animal to the human recipient is prevented, donor organ anatomy and function are compatible, and immunological rejections (HAR, VAR MAC) can be overcome. The most promising strategy is the synthesis of human complement regulatory proteins in the pig. Transgenic pigs have been generated for hDAF or hCD 46 and their hearts have been transplanted into non human primates for up to 90 days. HCD 59 driven by CMV promotor provides significant protection against HAR at least under in vitro and in vivo conditions. Current studies indicate a temporary use in patients within the next 3-5 years. As the microinjection technology to produce transgenic offspring is time consuming and very expensive, nuclear transfer technology provides a possibility for multiplication without going through the germ line with recombination effects. A functional nuclear transfer system will be crucial for xenotransplantation as it is anticipated that the expression of several transgenes will be required. PMID- 12654020 TI - Cyclic AMP signalling during mammalian sperm capacitation--still largely terra incognita. AB - Cyclic AMP is known to play a major role in intracellular signalling during mammalian sperm capacitation. However, despite much research, many of the molecular details of cyclic AMP's involvement remain obscure. In this review, I discuss the following aspects, presenting some original data as illustration where relevant. With respect to cyclic AMP synthesis, uncertainties exist as to the number of forms of adenylyl cyclase that are present in the spermatozoon, whether they are cytosolic or bound to subcellular structures, and which physiological effectors they respond to (e.g. bicarbonate, Ca2+, or receptor coupled G-proteins). While net intracellular levels of cyclic AMP in spermatozoa depend upon the relative activities of adenylyl cyclase and phosphodiesterase, there are wide between-sample variations within species, both in basal levels and in levels attained after activation of the cyclase (e.g. after sperm treatment with bicarbonate). Moreover, minor changes in bulk cyclic AMP levels can result in large changes in cyclic AMP-dependent functions. Finally, while cyclic AMP levels respond very rapidly to sperm treatment by effectors such as bicarbonate and Ca2+ (key components of capacitating media), there are big discrepancies between the rates of functional response. For example, enhancement of motility and collapse of phospholipid asymmetry take place within a few minutes, whereas more than 1 h of exposure to capacitating conditions is needed for cyclic AMP dependent protein tyrosine phosphorylation to become detectable or for the sperm population to attain a capacitated state. PMID- 12654019 TI - Resorption of the element zinc from spermatozoa by the epididymal epithelium. AB - In this study, elimination of the element zinc from spermatozoa during epididymal maturation was investigated. Testes and epididymides from 40 bulls were collected; epididymal fluid was flushed, pooled, labelled with 0.5 MBq 65Zn2+ per sample and proteins were separated on a Sephacryl S-200 HR and zinc chelate column chromatography. To follow the resorption of zinc in the epididymal epithelial lining, an autometallographic technique (AMG) was performed in tissue from caput, corpus, cauda and vas deferens. The results showed a zinc-binding protein fraction with an apparent molecular weight of 150-160 kDa, which was enriched after chelate column chromatography. Specific labelling of 65Zn was about five times higher in the caput than in the cauda epididymidis. AMG revealed no detectable zinc in the caput, but a significant increase of zinc resorption from the corpus to the cauda and vas deferens. Controls showed that the detectable zinc was located within the principal cells. In conclusion, our study proves that zinc present in the sperm flagellum starts to be mobilized in the caput epididymidis and is resorbed by the epididymal epithelium as from the corpus. This zinc elimination is a mandatory step in sperm maturation to obtain motility. PMID- 12654022 TI - Hyperactivated motility in sperm. AB - Hyperactivation is a movement pattern seen in sperm at the site and time of fertilization in mammals. It may be critical to the success of fertilization, because it enhances the ability of sperm to detach from the wall of the oviduct, to move around in the labyrinthine lumen of the oviduct, to penetrate mucous substances and, finally, to penetrate the zona pellucida of the oocyte. The movement of hyperactivated sperm appears different under different physical conditions and in different species, but basically it involves an increase in flagellar bend amplitude and, usually, beat asymmetry. Presumably, a signal or signals exist in the oviduct to initiate hyperactivation at the appropriate time; however, none has yet been identified with certainty. While the signal transduction cascade regulating hyperactivation remains to be completely described, it is clear that calcium ions interact with the axoneme of the flagellum to switch on hyperactivation. Although hyperactivation often occurs during the process of capacitation, the two events are regulated by somewhat different pathways. PMID- 12654021 TI - Cloning and characterization of boar epididymal secretory proteins by homology to the human. AB - Northern blot analysis suggested that the boar epididymis produces closely related counterparts to human epididymal proteins HE1, HE3, HE4, HE5 and HE12. 'Full-length' cloning by nucleic acid and amino acid sequence similarity was achieved by RT-PCR methods in the case of the porcine counterparts of HE3 and HE4, while the homologues of HE5 and HE12, despite their cross-hybridization during Northern blot analysis, have not yet been cloned. The two novel porcine cDNAs were derived from moderately abundant epididymal mRNAs that were 75 and 83% identical to HE3 and HE4 cDNAs, respectively. To emphasize their relationship to the corresponding HEs, they were named Se3 and Se4 cDNAs. Their open reading frames predicted small secretory proteins with 55% (Se3) and 76% (Se4) conserved amino acids. Monospecific antipeptide antibodies to HE secretory proteins identified He3- and HE12-related proteins on Western blots of porcine epididymal fluid and semen. Both Northern and Western analyses indicated that the Se proteins were produced in a regionalized pattern and accumulated in the cauda fluid. PMID- 12654023 TI - Seeking mathematical strategies in sperm function analysis: between scylla and charybdis? AB - During the last decades, essential progresses in reproductive biotechnology were achieved, implying development of special spermatological techniques. The major problem was to set up simple, rapid, precise and adequate evaluation methods. The key aspect to be considered in all assays of sperm fertilizing function is capacitation. As not all spermatozoa respond to fertilizing conditions in a similar manner, it seems to be logical to assess samples via their response to these specific conditions. For the spermatological practice, the sensitivity of methodology for assessment and analysis of data with respect to differences in individual response, in heterogeneity of the population, and proper temporal characterization of the response is crucial for the improvement of evaluation procedures. Currently, most used statistical analytical tools in spermatology do not always fulfil these essential sensitivity requirements. We structured our paper concerning different fields of mathematical science (distribution analysis, fractal geometry, functional approximation and differentiation) related to the modern insights in sperm function analysis. The spectrum of methods we are going to review in this paper is restricted to basic ideas to illustrate how the accuracy and sensitivity of sperm evaluation assays may be improved by applying adequate elementary tools of the mathematical analysis. PMID- 12654024 TI - Application of techniques for sperm selection in fresh and frozen-thawed stallion semen. AB - The objective of this research was to improve the techniques in processing chilled and frozen-thawed horse semen. In a preliminary experiment (Exp. I), different techniques for sperm selection and preparation [Swim-up, Glass wool (GW) filtration, Glass wool Sephadex (GWS) filtration; Percoll] were tested for their suitability for equine spermatozoa and results were compared with the routine procedure by dilution (Exp. I). In the main experiment (Exp. II), two sperm preparation techniques (GWS, Leucosorb) refering to the results of Exp. I and a previous study of our group (Pferdcheilkunde 1996 12, 773) were selected for processing complete ejaculates either for cooled-storage or cryopreservation. In a third experiment (Exp. III), pregnancy rates from inseminations with semen processed according to the techniques tested in Exp. II were compared with those obtained with semen processed according to routine procedures. In Exp. I (six stallions, six ejaculates/stallion), between 48 and 92% of spermatozoa were lost following the different sperm selection procedures (p < 0.05). Preparation of sperm increased percentage of progressively motile spermatozoa (pms) [Swim-up, GW, GWS vs dilution, Percoll (p < 0.05)] and decreased percentage of sperm head abnormalities [Swim-up, GW, GWS vs dilution, Percoll (p < 0.05)] probably by not improving the quality of individual cells, but by elimination of spermatozoa of inferior quality. In Exp. II (eight stallions, three ejaculates/stallion) Leucosorb and GWS procedures allowed the filtration of large volumes (extended ejaculates) for routine laboratory practice. GWS and Leucosorb filtration resulted in increased motility, membrane integrity and sperm viability after storage of spermatozoa until 48 h at +5 degrees C when compared with control (diluted) and centrifuged semen (p < 0.05). Significantly more spermatozoa were recovered after centrifugation (87.8 +/- 15.4%) compared with GWS (63.5 +/- 18.6%) and Leucosorb filtration (53.6 +/- 22.3%). GWS or Leucosorb procedure resulted in successful cryopreservation of stallion semen without centrifugation for removal of seminal plasma. The per cycle conception rate of inseminated mares using 200 x 106 pms transferred within 8 h after collection of semen was not affected by GWS filtration or Leucosorb separation when compared with centrifugation (n.s.; Exp. III). In conclusion, GWS and Leucosorb filtration results in the improvement of semen quality and should be considered as a method for stallion semen processing. Additional studies are needed for the evaluation of potentially higher fertilizing ability of stallion spermatozoa separated by techniques for sperm selection. PMID- 12654025 TI - On the species specificity of sperm binding and sperm penetration of the zona pellucida. AB - Sperm binding and sperm penetration of the zona pellucida (zp) are regarded as species-specific. In this investigation, the interactions between bovine oocytes and porcine, respectively, equine spermatozoa have been studied under in vitro conditions and compared with the normal in vitro fertilization of bovine oocytes by bovine sperm. Surprisingly, many of the heterologous spermatozoa adhered firmly to the bovine oocytes and could not be removed by intense washing. On average, more than 100 boar or equine spermatozoa were bound to the zp of bovine oocytes. Electron microscopic studies clearly demonstrated that porcine sperm attached to the zona and underwent the acrosome reaction. Equine spermatozoa displayed a similar binding affinity, but unlike the porcine spermatozoa even penetrated the zp and were taken up into the oocyte after a longer period of co incubation. Considering these new results the dogma of a strict species specificity of sperm zona interactions under in vitro conditions has to be reconsidered. PMID- 12654026 TI - Reflections upon sperm-endosalpingeal and sperm-zona pellucida interactions in vivo and in vitro. AB - In terms of experimental studies of the process of fertilization in mammals, this essay attempts to give a balanced assessment of current interpretations and to raise questions of direct relevance to two fields of research. First, the nature of pre-ovulatory sperm-endosalpingeal binding is set in a physiological perspective, and examination of this dynamic process in vitro is shown to suffer from serious shortcomings. In particular, sperm-epithelial binding reactions demonstrated in cultures of endosalpingeal cells may represent only a portion of the binding reaction in vivo or, as revealed by sperm binding to the tracheal epithelium, could be largely non-specific. Second, concerning the subsequent phase of binding of a fertilizing spermatozoon to the zona pellucida, the nature of the initial contact arrest of the sperm head is evaluated, especially in the light of the highly uneven or 'pitted' surface of this outer coating of the oocyte. Bearing in mind the active phase of oviduct macromolecular secretion that continues after ovulation, the question is posed as to how functional sperm binding sites are preserved on the surface of the zona pellucida rather than masked by epithelial and cumulus cell secretions. Finally, it is recalled that initial sperm : egg ratios at the site of fertilization in the ampullary-isthmic region of the oviduct are close to unity in diverse species of mammal, such fertilizing spermatozoa being released from prominent reserves in the caudal isthmus shortly before the time of ovulation. On strictly quantitative grounds, therefore, the extent to which biochemical or molecular studies portray changes in the vanguard of fertilizing spermatozoa needs to be questioned. Such studies are more likely to be descriptive of overall changes in very large numbers of sperm cells at diverse stages of maturation. PMID- 12654027 TI - Identification of cytokeratins in bovine sperm outer dense fibre fractions. AB - Outer dense fibres (ODF) are important substructures of mammalian sperm tails that are involved in the regulation of sperm motility. In this study, we investigated the identity of several sodium dodecyl sulphate (SDS)-insoluble ODF proteins. Bovine ODF were purified by separating sperm heads and tails using ultrasound and Percoll(R) density gradient centrifugation. Sperm flagella were treated with the detergent cetyltrimethylammonium bromide (CTAB). CTAB-insoluble material, which reportedly represents the ODF fraction, was collected, and electron microscopy confirmed a highly purified ODF fraction. We found after solubilization of this fraction with SDS that high amounts of insoluble material were retained after centrifugation. SDS-insoluble material was collected and quantitatively dissolved in 8 M urea. SDS-gel electrophoresis in the presence of urea revealed polypeptides with apparent molecular masses of approximately 25, 43, and 50 kDa. Subsequent immunoblotting with anti-cytokeratin antibodies detected two urea-soluble, SDS-insoluble proteins with apparent molecular masses of 45 and 66 kDa. The 45-kDa protein was identified as cytokeratin 19. An antibody reacting with a palette of cytokeratins (CK 1-18 and CK 20), KL1, was the only antibody that reacted with the 66-kDa polypeptide. We conclude that sperm ODF fractions contain at least one each of type I and type II intermediate filaments. As keratins and intermediate filaments are described as rope-like structures, we suggest that these intermediate filaments play an important structural or tension-bearing role in sperm flagella. PMID- 12654029 TI - Purine and pyrimidine biosynthesis in higher plants. AB - Purine and pyrimidine nucleotides have important functions in a multitude of biochemical and developmental processes during the life cycle of a plant. In higher plants the processes of nucleotide metabolism are poorly understood, but it is in principle accepted that nucleotides are essential constituents of fundamental biological functions. Despite of its significance, higher plant nucleotide metabolism has been poorly explored during the last 10-20 years (Suzuki and Takahashi 1977, Schubert 1986, Wagner and Backer 1992). But considerable progress was made on purine biosynthesis in nodules of ureide producing tropical legumes, where IMP-synthesis plays a dominant role in primary nitrogen metabolism (Atkins and Smith 2000, Smith and Atkins 2002). Besides these studies on tropical legumes, this review emphasises on progress made in analysing the function in planta of genes involved in purine and pyrimidine biosynthesis and their impact on metabolism and development. PMID- 12654028 TI - Assessing in vivo fertilizing capacity of liquid-preserved boar semen according to the 'Hanover gilt model'. AB - The goal of this study was to determine the ability of the Hanover gilt model to assess in vivo fertilizing capacity of preserved sperm and to consider whether any modifications to this model were needed. This model evaluates the fertilizing capacity of semen based on the fertilization rate, the rate of normal embryos and the accessory sperm count of 3-5-day embryos. Its distinguishing characteristics are the use of one-time insemination of sperm in reduced numbers, of spontaneously ovulating gilts and of ovulation detection through ultrasound examination of ovaries. Reduced sperm numbers allow for an accurate evaluation of the fertilizing potential of different semen treatments, thereby avoiding the compensatory effect of doses calibrated to maximize fertility. The model's usefulness was assessed in a trial run designed to compare the fertilizing capacity of liquid boar semen diluted into two different extenders. The diluent, the boar and the backflow, had no significant effect on any of the parameters studied. Gilts inseminated less than 24 h before ovulation had a significantly higher (p < 0.01) fertilization rate and accessory sperm cell count (p < 0.05) than those inseminated more than 24 h before ovulation. Very good/good embryos from homogeneous litters (only very good/good embryos were present) had a significantly higher (p < 0.01) accessory sperm count than those from heterogeneous litters (at least one embryo was of a different quality and/or oocytes were present). Both very good/good and degenerated/retarded embryos from heterogeneous litters had low accessory sperm numbers. This suggests that accessory sperm count is significantly related to the quality of the litter, but not to the quality of the embryo within gilts. It can be concluded that the Hanover gilt model is sensitive enough to show fertility differences (in this study, those associated with in vivo ageing of semen), while using relatively few gilts and little time. PMID- 12654030 TI - Phytochrome controlled signalling cascades in higher plants. AB - Plants can sense the changes in the environmental light conditions with highly specialized photoreceptors. Phytochromes are sensitive to red and far-red light and have a dual role in the life of plants. These photoreceptors play an important role in plant growth and development from germination to seed maturation and they are also involved in synchronizing the circadian clock with light/dark cycles. Biochemical, cell biological and genetic studies have been carried out to elucidate the molecular mechanism by which phytochromes transduce light signals. A major step in this process seems to be the light-dependent nuclear import of phytochromes. In the nuclei phytochromes interact with transcription factors and regulate the expression of numerous genes, resulting in complex physiological and developmental responses to light. This review focuses on the recently obtained results leading to the identification of some factors and processes involved in phytochrome signalling. PMID- 12654031 TI - Measurements of photosynthesis and respiration in plants. AB - Methods for measuring the rates of photosynthesis and respiration in plants are reviewed. Closed systems that involve manometric techniques, 14CO2 fixation, O2 electrodes and other methods for measuring dissolved and gas phase O2 are described. These methods typically provide time-integrated rate measurements, and limitations to their use are discussed. Open gas exchange systems that use infra red CO2 gas analysers and differential O2 analysers for measuring instantaneous rates of CO2 and O2 exchange are described. Important features of the analysers, design features of gas exchange systems, and sources of potential error are considered. The analysis of chlorophyll fluorescence parameters for estimating the quantum yield for O2 evolution and CO2 fixation is described in relation to new fluorescence imaging systems for large scale screening of photosynthetic phenotypes, and the microimaging of individual chloroplasts. PMID- 12654032 TI - Overexpression of alfalfa cytosolic glutamine synthetase in nodules and flowers of transgenic Lotus japonicus plants. AB - Legumes can obtain nitrogen from symbiotic nitrogen fixation in root nodules. The glutamine synthetase/glutamate synthase cycle is responsible for the initial nitrogen assimilation. This work reports the analysis of transgenic Lotus japonicus plants with the chimeric gene containing the alfalfa cytosolic glutamine synthetase (GS1) (EC 6.3.1.2) gene controlled by the Sesbania rostrata leghemoglobin gene promoter (Srglb3p). Surprisingly, all of the transgenic primary transformants analysed were sterile. Two transformants designated GS39 and GS44 were further analysed. GS in nodules of GS39 and GS44 plants was upregulated, at the level of transcript and protein. The transgenic plants had 2 fold higher nodule GS activity and similar root GS activity compared to control plants. The GS39 and GS44 sterile plants showed morphological alterations in pollen grains and in ovules. An increase in GS transcript abundance and enzyme activity was measured during early and late stages of flower development of GS plants. Flowers of GS plants showed higher glutamine content, resulting in an increased glutamine/glutamate ratio. The GS transcript and protein were detected in ovules. These data indicate that overexpression of GS1 in reproductive organs critically affects their development and might be a reason for sterility of L. japonicus plants. PMID- 12654033 TI - Aldehyde oxidase isoforms and subunit composition in roots of barley as affected by ammonium and nitrate. AB - Aldehyde oxidase (AO; EC 1.2.3.1) isoforms in roots of barley plants grown on ammonium or nitrate as nitrogen sources were studied. Roots of ammonium-grown barley plants exhibited considerable levels of AO2, AO3, and AO4 activities after native PAGE. Significantly lower AO2 and AO3 activity bands were observed in roots of plants grown on nitrate. When abscisic aldehyde was used as a substrate a strong response of the AO2 band was observed as well as a faint reaction of the AO3 band, but no activity of AO4 was observed using this substrate. The 160 and 145 kDa polypeptides were detected in ammonium grown plants. Root extracts of nitrate-fed plants revealed only a minor 145 kDa protein band and none of the 160 kDa subunit was detected. The assembly of the AO3 heterodimer requires the simultaneous presence of 160 and 145 kDa subunits. Subunit analysis of AO2 and AO4 revealed homodimeric composition of 160 and 145 kDa, respectively. Western blot analysis revealed changing AO subunits levels during germination and plant development. Differential expression of AO subunits (160 and 145 kDa) and subsequent formation of isoforms, which differ in substrate specificity, distribution and fulfil different enzymatic reactions, may constitute an important regulatory mechanism in the plant. PMID- 12654034 TI - Effects of season-dependent irradiance levels and nitrogen-deficiency on photosynthesis and photoinhibition in field-grown rice (Oryza sativa). AB - Photoinhibition and acclimation of photosynthesis in rice plants grown under N sufficient (NS) and N-deficient (ND) field conditions were investigated during the tropical wet (WS) and dry (DS) seasons in the Philippines. Diurnal patterns of CO2 assimilation were examined. There was a transient peak in CO2 assimilation in the leaves of the NS plants in the early morning during the DS and the WS, which was not seen in the ND plants in either season. ND leaves had lower Ribulose bisphosphate carboxylase/oxygenase (Rubisco) contents and lower chlorophyll contents. A lowered quantum yield of photosystem II (phiPSII) was observed in the ND plants at an intermediate irradiance though no differences between N treatments were seen at high irradiance. Analysis of carotenoids indicated a small increase in the de-epoxidation state of the xanthophyll cycle (DES) at mid-day in the ND leaves compared to NS. Photoinhibition was greater in ND leaves when incident mid-day irradiance was increased by altering the leaf angle. Although Rubisco contents were lower in ND plants, photosynthesis in situ did not decline proportionally. For NS plants, Chlorophyll content, but not Rubisco content, was season-dependent and results are discussed in terms of the interaction between irradiance use and N content of rice leaves. PMID- 12654035 TI - Monoterpene levels in needles of Douglas fir exposed to elevated CO2 and temperature. AB - Monoterpene levels in current year needles of Douglas fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings were measured at the end of 4 years of exposure to ambient or elevated CO2 (+179 micro mol mol-1), and ambient or elevated temperature (+0.3.5;C). Eleven monoterpenes were identified and quantified using gas chromatography/flame ionization detector/mass spectroscopy, with eight of these compounds regularly occurring in all trees examined. Elevated CO2 exposure significantly reduced the levels for four of the eight main compounds in needles. Total monoterpene production was reduced by 52% (P < 0.05). Elevated temperature also reduced monoterpene levels (P < 0.07). The combination of elevated temperature and elevated CO2 resulted in a 64% reduction in total monoterpenes compared with needles on ambient temperature trees. Two-way anova showed no significant temperature-CO2 interaction. It is hypothesized that seasonal reductions in needle monoterpene pools under elevated CO2 and temperature conditions may be due to a combination of competing carbon sinks, including increased carbon flux through the roots. PMID- 12654036 TI - Inhibition of growth and development of root border cells in wheat by Al. AB - The production and development of border cells vary with genotype, and they are released in wheat at an earlier stage of root development than other species studied so far. No significant difference was observed in the maximum number of border cells between Al-tolerant (Atlas 66) and Al-sensitive (Scout 66) cultivars in the absence of Al treatment. Al seriously inhibited the production and release of border cells, resulting in clumping of border cells in Scout 66, but less clustering in Atlas 66. The number of border cells released from roots treated with Al is significantly less than that from roots grown without Al treatment. Al treatment induced the death of detached border cells in vitro and they were killed by a 20-h treatment with 25 micro m Al. No significant difference in survival percentage of detached border cells was observed between Atlas 66 and Scout 66, regardless of the presence or absence of Al. The removal of border cells from root tips of both Atlas 66 and Scout 66 enhanced the Al-induced inhibition of root elongation concomitant with increased Al accumulation in the root. These results suggest that border cells adhered to the root tips play a potential role in the protection of root from Al injury in wheat. PMID- 12654037 TI - Fluoride inhibits root water transport and affects leaf expansion and gas exchange in aspen (Populus tremuloides) seedlings. AB - The effects of sodium fluoride (0.3, 5 and 10 mM NaF) on root hydraulic conductivity, and gas exchange processes were examined in aspen (Populus tremuloides Michx.) seedlings grown in solution culture. A long-term exposure of roots to NaF significantly decreased root hydraulic conductivity (Lp) and stomatal conductance ( gs). Root absorbed NaF significantly affected electrolyte leakage in leaf tissues and substantially restricted leaf expansion. NaF did not significantly affect leaf chlorophyll contents but decreased net photosynthesis (Pn). A short-term exposure of excised roots to 5 mM NaF and KF significantly decreased root water flow (Qv) with a concomitant decline in root respiration and reduced gs when applied through intact roots or excised stems. The same molar concentration of NaCl also decreased Qv and gs in intact seedlings, but to a lesser extent than NaF or KF, and did not significantly affect root respiration. The results suggest that fluoride metabolically inhibited Qv or Lp, probably by affecting water channel activity. We suggest that the metabolic inhibition of Lp by root-absorbed fluoride affected gas exchange and leaf expansion in aspen seedlings. PMID- 12654038 TI - Light-dependent photosynthetic characteristics indicated by chlorophyll fluorescence in five mangrove species native to Pohnpei Island, Micronesia. AB - To investigate the adaptive capacity to strong light among mangrove species, we examined light-dependent properties of photosynthesis in relation to photoinhibition using chlorophyll fluorescence for sun-leaves of five mangrove tree species; Sonneratia alba, Rhizophora stylosa, Rhizophora apiculata, Bruguiera gymnorrhiza and Xylocarpus granatum. These species were classified into three groups; pioneer -S. alba and R. stylosa, intermediate -R. apiculata and B. gymnorrhiza and climax species -X. granatum. A further distinction can be made between the two intermediate species, the less shade-tolerant R. apiculata and the shade-tolerant B. gymnorrhiza. There was a clear association between successional status and light-saturated electron transport rate (ETR) where ETR was higher in S. alba and R. stylosa > R. apiculata and B. gymnorrhiza > X. granatum. Based on its lower initial slope of light-dependent ETR and lower ratio of variable to maximum fluorescence (Fv/Fm) after a dark-adaptation, X. granatum appeared to suffer greater photoinhibition than other species. The response of qP, which represents PSII openness, to incident light indicates a ranking of tolerant capacity to photoinhibition of S. alba and R. stylosa > R. apiculata > B. gymnorrhiza and X. granatum. The difference in qP response between R. apiculata and B. gymnorrhiza might be attributed to the higher leaf absorptance and the lower degree of thermal dissipation indicated by the higher F'v/F'm in B. gymnorrhiza. It appears that the co-ordination of leaf absorptance, thermal dissipation and consumption of absorbed light energy through electron transport is important in the acclimation of mangrove species to exposed habitats. PMID- 12654039 TI - Longitudinal leaf gradients of UV-absorbing screening pigments in barley (Hordeum vulgare). AB - In vivo leaf characteristics were examined to describe longitudinal gradients of UV-absorbing screening pigments in barley. Chlorophyll fluorescence properties and in vivo absorption spectra (210-750 nm) of leaves were measured from the base to the tip. Barley leaves showed strong longitudinal gradients of chlorophyll, where chlorophyll concentration increased within the first 5-8 cm from the leaf base, and did not significantly change for the remaining part of the leaf. Fluorescence microscopy was used to localize cell wall bound screening pigments different from flavonoids, since flavonoids lack a blue-green fluorescence emission (Lichtenthaler and Schweiger 1998). Measurements of in vivo chlorophyll fluorescence indicated that the ratio of UV-absorbing screening pigments per leaf area increases from the leaf base to the tip. These gradients were confirmed by in vivo absorption spectra. It is demonstrated that leaves in the early stage of development are less protected against UV-radiation than fully developed mature leaf regions. The experiments show that measurements of in vivo chlorophyll fluorescence are ideally suited as a fast non-invasive tool to estimate the epidermal UV-transmittance in different leaf sections. PMID- 12654040 TI - The impact of elevated ozone and carbon dioxide on young Acer saccharum seedlings. AB - The effects of high O3 (200 nl l-1 during the light period) and high CO2 (650 &mgr;l l-1 CO2, 24 h a day) alone and in combination were studied on 45-day-old sugar maple (Acer saccharum Marsh.) seedlings for 61 days in growth chambers. After 2 months of treatment under the environmental conditions of the experiment, sugar maple seedlings did not show a marked response to the elevated CO2 treatment: the effect of high CO2 on biomass was only detected in the leaves which developed during the treatment, and assimilation rate was not increased. Under high O3 at ambient CO2, assimilation rate at days 41 and 55 and Rubisco content at day 61 decreased in the first pair of leaves; total biomass was reduced by 43%. In these seedlings large increases (more than 2-fold) in glucose 6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) activity and in anaplerotic CO2 fixation by phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) were observed, suggesting that an enhanced reducing power and carbon skeleton production was needed for detoxification and repair of oxidative damage. Under high O3 at elevated CO2, a stimulation of net CO2 assimilation was observed after 41 days but was no longer observed at day 55. However, at day 61, the total biomass was only reduced by 21% and stimulation of G6PDH and PEPC was less pronounced than under high O3 at ambient CO2. This suggests that high CO2 concentration protects, to some extent, against O3 by providing additional carbon and energy through increased net assimilation. PMID- 12654041 TI - Synergistic interactions between volicitin, jasmonic acid and ethylene mediate insect-induced volatile emission in Zea mays. AB - Plants display differential responses following mechanical damage and insect herbivory. Both caterpillar attack and the application of caterpillar oral secretions (OS) to wounded leaves stimulates volatile emission above mechanical damage alone. Volicitin (N-17-hydroxylinolenoyl-l-glutamine), present in beet armyworm (BAW, Spodoptera exigua) OS, is a powerful elicitor of volatiles in excised maize seedlings (Zea mays cv. Delprim). We consider some of the mechanistic differences between wounding and insect herbivory in maize by examining the activity of volicitin, changes in jasmonic acid (JA) levels, and volatile emission from both intact plant and excised leaf bioassays. Compared to mechanical damage alone, volicitin stimulated increases in both JA levels and sesquiterpene volatiles when applied to intact plants. In a bioassay comparison, excised leaves were more sensitive and produced far greater volatile responses than intact plants following applications of both volicitin and JA. In the excised leaf bioassay, volicitin applications (10-500 pmol) to wounded leaves resulted in dose dependent JA increases and a direct positive relationship between JA and sesquiterpene volatile emission. Interestingly, volicitin-induced JA levels did not differ between intact and excised bioassays, suggesting a possible interaction of JA with other regulatory signals in excised plants. In addition to JA, insect herbivory is known to stimulate the production of ethylene. Significant increases in ethylene were induced only by BAW herbivory and not by either wounding or volicitin treatments. Using intact plant bioassays, ethylene (at 1 micro l l-1 or less) greatly promoted volatile emission induced by volicitin and JA but not mechanical damage alone. For intact plants, wounding, elicitor-induced JA and insect-induced ethylene appear to be important interacting components in the stimulation of insect-induced volatile emission. PMID- 12654042 TI - Aquaporin functionality in relation to H+-ATPase activity in root cells of Capsicum annuum grown under salinity. AB - As water and nutrient uptake should be related in the response of plants to salinity, the aim of this paper is to establish whether or not aquaporin functionality is related to H+-ATPase activity in root cells of pepper (Capsicum annuum L.) plants. Thus, H+-ATPase activity was measured in plasma membrane vesicles isolated from roots and aquaporin functionality was measured using a cell pressure probe in intact roots. Salinity was applied as 60 mM NaCl or 60 mM KCl, to determine which ion (Na+, K+ or Cl-) is producing the effects. We also investigated whether the effects of both salts were ameliorated by Ca2+. Similar results were obtained for cell hydraulic conductivity, Lpc, and H+-ATPase activity, large reductions in the presence at NaCl or KCl and an ameliorative effect of Ca2+. However, fusicoccin (an activator of H+-ATPase) did not alter osmotic water permeability of protoplasts isolated from roots. Addition of Hg2+ inhibited both ATPase and aquaporins, but ATPase also contains Hg-binding sites. Therefore, the results indicate that H+-ATPase and aquaporin activities may not be related in pepper plants. PMID- 12654043 TI - Casparian strips in needles of Pinus bungeana: isolation and chemical characterization. AB - By using cell wall degrading enzymes, Casparian strips were for the first time isolated from Pinus bungeana needle endodermis. They appeared as a fine network, similar to those isolated from roots. Fourier transform infrared spectroscopic analysis provided evidence that the Casparian strips were impregnated with lignin, suberin, cellulose and cell wall proteins. PMID- 12654044 TI - Cloning of a pine germin-like protein (GLP) gene promoter and analysis of its activity in transgenic tobacco Bright Yellow 2 cells. AB - Germins and germin-like proteins (GLPs) constitute a large and highly diverse family of ubiquitous plant cell wall proteins. These proteins seem to be involved in many developmental stages and stress-related processes, but their exact participation in these processes generally remains obscure. In Pinus caribaea Morelet, the PcGER1 gene is expressed uniquely in embryo tissues, and encodes a GLP ionically bound to the walls of pine embryo cells maintained in 2,4-D containing medium. We have cloned a genomic fragment including the 1520 bp 5' upstream promoter region of PcGER1. This sequence contains, in its 1200 bp distal part, several cis elements (e.g. SEF4, 60 kDa protein, ABA RE and Dof recognition sites) present in genes responding to hormones and/or expressed in embryo or seed tissues, or during germination. The PcGER1 promoter sequence was cloned upstream of the GUS (beta-glucuronidase) reporter gene and transferred to tobacco Bright Yellow 2 (BY-2) cells via Agrobacterium tumefaciens-mediated transformation. Promoter activity and growth performances of transgenic asynchronous cell suspensions were analysed in the presence or absence of 2,4-D and/or BA. Optimal growth, maximum cell-wall yield and PcGER1 promoter activity were observed in the presence of 2,4-D and BA at day 4, the end of the exponential growth phase where 70-75% cells have a 2C DNA content. Analysis of promoter activity during the cell cycle in an aphidicoline-synchronized culture suggested that the expression is maximum in G1 cells. We also showed that under optimal growth conditions, 5' promoter deletions decreased the activity of the reporter gene. We discuss the function of this gene with regards to cell growth. Accession number: The PcGER1 promoter sequence was submitted to the genbank database under the accession number AY077704. PMID- 12654045 TI - Cuticle characteristics and volatile emissions of petals in Antirrhinum majus. AB - Floral volatiles, which are small and generally water-insoluble, must move from their intracellular sites of synthesis through the outermost cuticle membrane before release from the flower surface. To determine whether petal cuticle might influence volatile emissions, we performed the first analysis of petal cuticle development and its association with the emission of flower volatiles using Antirrhinum majus L. (snapdragon) as a model system. Petal cuticular wax amount and composition, cuticle thickness and ultrastructure, and the amounts of internal and emitted methylbenzoate (the major snapdragon floral scent compound) were examined during 12 days, from flower opening to senescence. Normal (n-) alkanes were found to be the major wax class of snapdragon petals (29.0% to 34.3%) throughout the 12 days examined. Besides n-alkanes, snapdragon petals possessed significant amounts of methyl branched alkanes (23.6-27.8%) and hydroxy esters (12.0-14.0%). Hydroxy esters have not been previously reported in plants. Changes in amount of methylbenzoate inside the petals followed closely with levels of methylbenzoate emission, suggesting that snapdragon petal cuticle may provide little diffusive resistance to volatile emissions. Moreover, clear associations did not exist between methylbenzoate emission and the cuticle properties examined during development. Nevertheless, the unique wax composition of snapdragon petal cuticles shows similarities with those of other highly permeable cuticles, suggesting an adaptation that could permit rapid volatile emission by scented flowers. PMID- 12654046 TI - Screening for GALC to make neonatal diagnosis and initial neonatal stem cell treatment with umbilical cord blood. PMID- 12654047 TI - Ambulatory blood pressure monitoring in pediatric renal transplantation. AB - Over last two decades ABPM has evolved from a research device to an established and valuable clinical tool for BP evaluation. More than 10 yrs ago ABPM was introduced to pediatrics and since that time, its importance has been increasing in the management of hypertension in children and adolescents. This review summarizes the information gathered from the studies of ABPM in adult and pediatric patients with renal transplants. We will review the importance of hypertension in this patient subset, discuss the advantage of ABPM over CBP and focus on specific abnormalities and clinical significance of ABPM in renal transplant recipients. PMID- 12654048 TI - Liver transplantation for cholestasis associated with cystic fibrosis in the pediatric population. AB - The most common hepatic complications of cystic fibrosis (CF) are steatosis, fibrosis, biliary cirrhosis, atretic gallbladder, cholelithiasis, and sclerosing cholangitis. Cholestatic liver disease is a slow progressive disorder, but will stabilize for many patients. CF patients may suffer from the consequences of their liver disease and without liver transplantation, variceal hemorrhage, malnutrition, or end-stage liver disease can lead to death. Prospective data were collected and reviewed on 311 liver transplants performed in 283 patients at the Children's Medical Center of Dallas between October 1984 and November 2000. Ten children received an orthotopic liver transplant (OTLX) for end-stage liver disease associated with cystic fibrosis. Pulmonary function tests were obtained preoperatively in all cases. There were nine boys and one girl. Six are currently alive, and four are dead. Both patient and graft survival was 5.75 yr. Among those currently alive, mean patient and graft survival is 7.71 yr (range 0.10 12.62 yr). Mean patient and graft survival of those who died was 2.35 yr (range 0.78-5.33 yr). No survivor required re-transplantation and currently, all have normal serum aminotransferase values. Chronic sinusitis was not a significant pre or post-transplant morbidity, although systematic radiographic evaluation of the sinuses did not occur. Pulmonary deaths occurred in three patients from pulmonary hemorrhage, pulmonary infection with Aspergillus and Candida glabrata, and acute bronchopneumonia associated with polymicrobial sepsis because of Pseudomonas, Klebsiella, and Candida albicans 1.44, 0.78, and 1.83 yr, respectively, after transplantation. The fourth death was associated with chronic rejection, and occurred 5.33 yr after transplantation. All non-survivors were below the 5th percentile for height and weight at the time of liver transplantation. Mean age at transplantation was 9.72 yr (range 1.23-19.09, median 9.61). Survivors were transplanted at a younger age than non-survivors (mean of 9.21 yr vs. 10.66 yr), and had shorter waiting times from diagnosis of end-stage liver disease to transplantation (6.87 months vs. 13.83 months). Eighty percentage (n = 8) of patients had pretransplant variceal bleeds (83% of survivors, 75% of non survivors). While all non-survivors had a history of meconium ileus and preoperative need of pancreatic enzymes, only 67% of those alive experienced these complications. Preoperative forced vital capacity FVC was 103% for survivors and 95% for non-survivors. The corresponding numbers for forced expiratory flow (FEF) 25-75 were 74-84% respectively. Preoperative Aspergillus was identified in 30% of patients (n = 3). Two of these patients are alive. Cystic fibrosis constitutes an indication for 3.5% of pediatric liver transplants. Evaluation and transplantation for end-stage liver disease associated with cystic fibrosis should be undertaken at an early age. Most deaths were associated with pulmonary/septic events, and occurred less than 2 yr after OLTX. Those children who did not survive had poor growth and nutrition, prolonged waiting times prior to transplantation, were transplanted at an older age, and had a higher incidence of pancreatic insufficiency and meconium ileus. The presence of Aspergillus in the sputum does not constitute a contraindication for OLTX. PMID- 12654049 TI - Hypophosphatemia and hypouricemia in pediatric allogeneic bone marrow transplant recipients. AB - Increased phosphate (P) uptake by the replicating neutrophils during engraftment syndrome has been described to play a role in the development of hypophosphatemia (HP) in bone marrow transplantation patients, and suggested as a measure of neutrophil recovery. Here, the relationship of serum P with engraftment was determined in 56 children who underwent allogeneic bone marrow transplantation (BMT). Uric acid (UA) levels were also analyzed to study the contribution of cytolysis on P levels. HP and hypouricemia (HU) developed at least once in 63 and 57% of patients respectively, before and until day +20 after transplantation. The minimal values of P and UA were observed at day +10 and were significantly lower than the baseline values (p < 0.01). The mean neutrophil engraftment was at day +13, following the P and UA nadir by 3 days. In addition there was a significant correlation between P and UA levels (p = 0.01). The levels of of both P and UA returned to pretransplant values at day +20. A significant correlation (p < 0.05) between platelet engraftment and P levels was also demonstrated. HP and HU seen in pediatric patients undergoing BMT reflects a combination of pathophysiologic mechanisms. PMID- 12654051 TI - Neurocognitive outcome in pediatric liver transplant recipients. AB - Studies have shown that children with liver transplants demonstrate deficits on intellectual, academic, and language measures. However, limited information is known about the long-term cognitive development of these children. In this study, 15 children who were at least 2 yr post-liver transplantation (LT) were compared on cognitive measures to an equated group of 15 children with cystic fibrosis (CF). Children with CF were selected as a clinical control given similarities in disease onset and chronicity, as well as physical growth and development. Results indicated that children with LT tended to have lower verbal intelligence quotient scores and performed significantly lower than CF children on language measures, particularly on receptive language tasks. No significant differences were obtained on measures of academic achievement or visual-spatial performance. In the LT group, days in the intensive care unit, total number of days spent in the hospital during the first year following the transplant, and elevated pretransplant bilirubin levels significantly predicted the speech and language delays. PMID- 12654050 TI - Predictive value of pretransplantation cyclosporine pharmacokinetic studies on initial post-transplantation dosing in pediatric kidney allograft recipients. AB - Despite the introduction of a variety of new immunosuppressive agents, cyclosporine A (CsA) has maintained a strong position in pediatric transplantation (Tx). Post-Tx dosing with CsA is a challenging task because of the narrow therapeutic window of the drug, the great individual variability of metabolism and the lack of consensus about the optimal dosage and targeted blood concentration. Sufficient administration of CsA may be protective against acute rejections and other early complications after Tx, which is crucial for the long term survival of the graft. Individual doses based on pre-Tx pharmacokinetic studies might be helpful in achieving optimal early concentrations of CsA. To asses the usefulness of pharmacokinetic studies, we retrospectively compared the post-Tx doses administered with the individually predicted doses between 1988 and 1998. Multiple regression of data on 65 de novo renal transplant recipients, 1.1 15.5 yr old, was used to analyze the significance of the predicted dose, trough blood concentration of CsA (B-CsA), serum creatinine and age at the time of Tx in explaining the doses used during the first three post-Tx weeks. Patients were grouped according to age (<2, 2-8 and >8 yr), according to the predicted dose (within or outside +/-25% of age-group average), and according to the oral formulation of CsA. Standard dosing scheme was simulated by using age-specific average doses in the place of the individual predicted doses. Administered doses of CsA were high [averaging 22.6 (504), 20.7 (484), and 12.4 mg/kg/d (329 mg/m2/d) for patients <2, 2-8, and >8 yr old] but the average B-CsA remained in the target range of 250-450 microg/L. The predicted dose and age were significant parameters in explaining the administered doses during the first 3 wk after Tx. B CsA and S-creatinine were non-significant. The predicted doses were used to initiate the dosing of CsA after Tx (R2 = 0.70) and adjustments to dosing were made during the next weeks (R2 = 0.59, 0.52). Multiple regression model showed better fit for 60% of our patients, who had atypical predicted doses (R2 = 0.74, 0.60, 0.64 for first, second and third post-Tx weeks, respectively), most remarkably in patients <2 yr of age, than for the study population as a whole. A simulated standard dose was not able to explain the administered doses of CsA. In conclusion, pre-Tx pharmacokinetic studies are valuable for determining individual post-Tx starting doses, especially for those patients who need high or low doses of CsA. Individual dosing led to relatively high initial CsA doses, which could be significant for the long-term survival of the graft. PMID- 12654052 TI - Tacrolimus-based immunosuppression with steroid withdrawal in pediatric kidney transplantation--4-year experience at a moderate-volume center. AB - Tacrolimus-based immunosuppression with steroid withdrawal in pediatric kidney transplantation was pioneered at the University of Pittsburgh but is not broadly practiced at other centers. We present our 4-year experience with a modified Pittsburgh protocol at our own moderate-volume center. Seventeen pediatric kidney transplant recipients were treated with a tacrolimus-based immunosuppressive regimen involving steroid withdrawal over 6-12 post-transplant months in most cases and followed for up to 49 months. Patient and graft survival as well as graft function were excellent, and beneficial effects on several cardiovascular parameters were noted. Complications included fungal infections, glucose intolerance and post-transplant lymphoproliferative disease and were generally managed successfully. While awaiting longer-term follow-up data, we conclude that tacrolimus-based immunosuppression with steroid withdrawal in pediatric kidney transplantation is potentially beneficial and feasible even at centers managing a rather small number of recipients. PMID- 12654053 TI - Prevalence and clinical significance of human herpesviruses 6 and 7 active infection in pediatric liver transplant patients. AB - Recent studies in adult liver transplant patients have suggested that both human herpesvirus (HHV)-6 and HHV-7 infection are important causes of morbidity following liver transplantation. However, the impact of HHV-6 and -7 infection in pediatric liver transplant patients remains largely unknown. The aims were to determine the prevalence of HHV-6 and -7 infection in pediatric liver transplant patients and to determine whether there is an association between HHV-6 and -7 infection with episodes of graft rejection and cytomegalovirus (CMV) infection. A total of 46 pediatric liver transplant patients transplanted at Mayo Clinic between January 1994 and January 2000 were evaluated. Quantitative polymerase chain reaction (PCR) assays for CMV, HHV-6 and HHV-7 were performed on stored sera obtained prior to transplant, weekly for 8 wk and at 4 months and 1 yr post transplant. Pretransplant sera were tested for HHV-6 antibodies by indirect immunofluorescence assay. A total of 215 blood samples were tested (mean 6.5 +/- 3.1, range 3-18). CMV infection occurred in 11 of 33 (33.3%) patients, while CMV disease occurred in 4 of 33 (12%) patients. Infection with HHV-6 (variant B) was detected in three of 33 (9.1%) patients. HHV-7 infection was not detected. Case 1 and 2 were infants (10- and 11-month old, respectively). Both were seronegative for HHV-6 pretransplant. In both cases, HHV-6 infection was associated with concurrent episodes of moderate to severe acute graft rejection. Case 3 was a 16 yr-old girl who was seropositive for HHV-6 pretransplant. No clinical events were recorded and a liver biopsy performed per protocol showed no evidence of rejection. None of the three patients had concomitant CMV infection or disease. In this study, HHV-6 infection occurred in 9% of pediatric liver transplant patients while HHV-7 was not detected. A potential association between primary HHV-6 infection and allograft rejection warrants further investigation. PMID- 12654055 TI - Mycophenolate mofetil without antibody induction in cadaver vs. living donor pediatric renal transplantation. AB - Mycophenolate mofetil (MMF) is a new immunosuppressive agent that blocks de novo purine synthesis in T and B lymphocytes via a potent selective inhibition of inosine monophosphate dehydrogenase. MMF has been shown to significantly reduce the incidence of acute rejection in both adult and pediatric renal transplantation. The impact of MMF on routine antibody induction therapy in pediatric renal transplantation has not been defined. Remarkably, a recent North American Pediatric Transplant Cooperative Study concluded that T-cell antibody induction therapy was deleterious for patients who received MMF. Our study examines the use of MMF in an evolving immunosuppressive strategy to avoid antibody induction in both living (LD) and cadaver (CAD) donor pediatric renal transplantation. We retrospectively analyzed the records of 43 pediatric renal transplants that received MMF-based triple therapy without antibody induction therapy between November 1996 and April 2000. We compared CAD (n = 17) with LD (n = 26). The two groups were similar demographically except that CAD had significantly younger donors than LD, 26.1 +/- 13.7 vs. 36.2 +/- 9.2 yr (p = 0.006). All the patients received MMF at 600 mg/m2/b.i.d. (maximum dose of 2 g/d) and prednisone with cyclosporine (86%) or tacrolimus (14%). Mean follow-up was >36 months for each group. Acute rejection rate at 6 months was 11.8% (CAD) vs. 15.4% (LD) (p = 0.999) and at 1 yr was 23.5% (CAD) vs. 26.9% (LD) (p = 0.999). Mean estimated glomerular filtration rate (ml/min/1.73 m2) at 6 months was 73.3 +/- 15.3 (CAD) vs. 87.6 +/- 24.2 (LD) (p = 0.068). Patient survival at 1, 2, and 3 yr was 100, 100, and 100% for CAD vs. 100, 96, and 96% for LD, respectively. Graft survival at 1, 2, and 3 yr was 100, 100, and 94% for CAD vs. 96, 88, and 71% for LD, respectively. Graft loss in CAD was because of chronic rejection (n = 2) while in LD it was because of non-compliance (n = 6), post-transplant lymphoproliferative disorder (n = 1), and sepsis (n = 1). In conclusion, MMF without antibody induction in both CAD and LD pediatric renal transplantation provides statistically similar and effective prophylaxis against acute rejection at 6 months and 1 yr post-transplant. The short-term patient and graft survival rates are excellent, however, non-compliance remains a serious challenge to long term graft survival. Additional controlled studies are needed to define the role of MMF without antibody induction therapy in pediatric renal transplantation. PMID- 12654054 TI - Risk factors for moderate-to-severe acute graft-vs.-host disease after allogeneic stem cell transplantation in children. AB - Severe acute graft-vs.-host disease (aGVHD) remains a major cause of transplantation-related mortality. However, because of a graft-vs.-leukaemia effect, a mild (grade I) aGVHD is desirable. As risk factors predisposing for aGVHD are not necessarily the same in children and adults, we have performed a retrospective analysis of risk factors (RFs) for grade II-IV aGVHD in 258 paediatric patients undergoing allogeneic stem cell transplantation at our centre. Thirty-two potential RFs were assessed with univariate analysis in logistic regression. Eleven factors were selected for further evaluation in stepwise elimination multivariate analysis. Three independent RFs were found: (1) donor other than human lekocyte antigen (HLA)-identical sibling [odds ratio (OR) 6.1, p < 0.001); (2) single drug [cyclosporine A (CsA) or methotrexate (Mtx)] graft-vs.-host disease (GVHD) prophylaxis (OR 7.0, p < 0.001); and (3) ABO disparity of any kind (OR 2.4, p = 0.02). The RFs were additive: moderate-to severe aGVHD was seen in none of the patients without any RFs; in 16% with one RF; in 32% with two RFs and in 67% with all three RFs present. Single drug GVHD prophylaxis (CsA or Mtx), any kind of ABO mismatch, and non-sibling donors are RFs for grade II-IV acute GVHD in paediatric SCT. We encourage the use of combination GVHD prophylaxis in children. ABO mismatch should be considered when choosing between otherwise equally suitable donors. PMID- 12654056 TI - The effect of aprotinin on transfusion requirements in pediatric orthotopic liver transplantation. AB - The use of aprotinin to reduce blood loss by inhibiting fibrinolysis thereby decreasing transfusion requirements during orthotopic liver transplantation (OLT), is well-documented in adults. We set out to test the hypothesis that the prophylactic use of aprotinin reduced blood product requirements during pediatric OLT. A retrospective study was performed, reviewing data from 24 OLTs performed over a 4-yr period. Six patients did not receive aprotinin (group 1), while 18 (group 2) received a weight-based dose of aprotinin. Both groups were comparable with respect to demographics, baseline characteristics and surgical variables except for a significantly more prolonged activated partial thromboplastin time (APTT) in the aprotinin group (p = 0.015). Despite the fact that median values for transfused volumes of red blood cells (78.3 vs. 36.7 mL/kg) and fresh frozen plasma (51.9 vs. 23.7 mL/kg) were more than halved in the aprotinin group, there was no statistical difference demonstrated. The failure to reach statistical significance can probably be explained by the small number in group 1 and a high level of scatter. All patients in group 1 required intraoperative transfusion of RBC and fresh frozen plasma (FFP) while two patients in group 2 did not require RBC and seven received no FFP. There were four patients in group 1 and 17 in group 2 who did not receive platelets while five in group 1 and 12 in group 2 did not receive cryoprecipitate. The differences between the groups in avoidance of these blood products did not reach statistical significance. There was little difference between groups with respect to albumin and crystalloid requirements. No statistical difference was demonstrated in intraoperative hematologic profiles between the two groups except during the anhepatic phase of surgery when there was a statistically significant more prolonged prothrombin time (p = 0.04) and a greater international normalized ratio (p = 0.027) in group 2. PMID- 12654058 TI - Treatment of monomorphic B-cell lymphoma with rituximab after liver transplantation in a child. AB - Rituximab, a monoclonal antibody directed against the B-cell specific CD20 antigen has been used with success in post-transplant lymphoproliferative disorder (PTLD) of B-cell phenotype. However, the use of such drug in children with liver transplantation and PTLD is very limited. We report a 2-yr-old liver transplant recipient with monomorphic non-Hodgkin lymphoma of B-cell origin. The lymphoma did not respond to immunosuppression withdrawal, with a subsequent allograft rejection. Despite resumption of immunosuppression and rejection treatment, the lymphoma was successfully treated with rituximab. PMID- 12654057 TI - Clinical approach to graft hepatic artery thrombosis following living related liver transplantation. AB - Hepatic artery thrombosis (HAT) has an occurrence rate of 1.7-26% following living donor liver transplantation (LDLT) and is one of the most common reasons for graft loss and mortality in this population. There is a higher incidence of HAT in pediatric recipients. The aim of this case report is to discuss clinical approaches for the treatment of HAT occurring in the early post-operative period after LDLT. An 11-month-old, 7.8-kg female with cirrhosis secondary to biliary atresia underwent LDLT at Gazi University Hospital in Ankara. The graft was a left lateral segment from her father with a left hepatic artery (HA) of 2 mm diameter and a graft weight/recipient body weight ratio of 2.0%. After an uneventful early post-operative period, HAT was diagnosed by Doppler ultrasonography (USG) on the fifth post-operative day. Following angiographic evaluation, immediate exploration and reanastomosis was performed using an operation microscope. Post-operatively, the HA was patented by Doppler USG and graft function returned to normal. Now, 42 months later, the patient continues to do well with normal graft function, using a regimen of tacrolimus monotherapy for immunosuppression. In countries which have very limited resources for urgent re transplantation, given their serious donor shortage, graft salvage may be the only option for patient survival when HAT occurs. In these circumstances, early diagnosis and immediate revascularization may be the only method for graft salvage. A daily routine of Doppler USG examination in the early post-operative period may provide a method for the early diagnosis of HAT, before liver enzymes are elevated and hepatic necrosis has begun. PMID- 12654060 TI - Pathologic changes of glial cells in murine model of Niemann-Pick disease type C: immunohistochemical, lectin-histochemical and ultrastructural observations. AB - BACKGROUND: In recent years, morbid states of glial cells have been reported in several neurodegenerative diseases. We studied neuropathologically the glial cells in a murine model of Niemann-Pick disease type C (NPC) to clarify involvement of glias, the most important supportive cells in the central nervous system, by the disease. METHODS: The brains of sphingomyelinosis mice (spm/spm), aged from 5 to 13 weeks, and 15 of their age-matched normal siblings were studied histopathologically, immunohistochemically and electron micro-scopically. RESULTS: Accumulation of ubiquitin-positive materials was found in the cytoplasm of foam cells and ballooned neurons immunohistochemically. In addition to the morphologically abnormal cells, double immunostaining of ubiquitin and glial fibrillary acidic protein (GFAP) revealed the deposition of ubiquitinated substances in the cytoplasm of astrocytes. Ultrastructurally, numerous concentric lamellar inclusions, so-called 'myelin figures', appeared in the neurons and phagocytotic cells. Some oligodendrocytes also contained 'myelin figure' inclusions and multivesicular inclusions. Astrocytes contained abnormal irregularily-shaped electron dense materials. CONCLUSIONS: In the murine model of NPC, astrocytes and oligodendrocytes are also involved in the morbid processes. Thus, it might be relevant to investigate the glial dysfunction to understand the pathological processes of the disease and to prepare an adjunct therapeutic strategy to manage the patients with NPC. PMID- 12654059 TI - Improvement of hepatopulmonary syndrome after transjugular intrahepatic portasystemic shunting: case report and review of literature. AB - The hepatopulmonary syndrome has been described in as many as 5-29% of patients with liver disease. Patients with this syndrome may suffer from chronic hypoxemia, and mortality rates of liver patients with this syndrome are as high as 41%. Early diagnosis of such patients is essential. Currently, liver transplantation is the only effective therapy for such patients, and reversal of this syndrome is seen in up to 80% of patients post-transplant. Transjugular intrahepatic portasystemic shunting (TIPS) as a therapeutic maneuver for this syndrome has been described in five patients to date with mixed results. Reduction in portal hypertension with consequent redistribution of blood flow and altered synthesis of vasodilatory chemicals have been postulated to help resolve this disease. In this report, we describe an 11-yr-old female with biliary atresia and hepatopulmonary syndrome. Her disease was complicated with recurrent variceal bleeding. TIPS achieved a therapeutic response of both her bleeding and respiratory complications. PMID- 12654061 TI - Intravenous glycerol therapy should not be used in patients with unrecognized fructose-1,6-bisphosphatase deficiency. AB - BACKGROUND: In Asian countries, glycerol solution that contains fructose (5%) is often used for management of brain edema. However, glycerol and fructose may cause severe hypoglycemia and metabolic acidosis in patients with fructose-1,6 bisphosphatase (FBPase) deficiency, even under stable conditions. The aim of the present study was to determine whether glycerol solution was used for brain edema during acute metabolic decompensation of hypoglycemia and metabolic acidosis in patients with unrecognized FBPase deficiency in Japan and to examine a long-term prognosis of the patients who had this kind of severe metabolic decompensation with or without glycerol therapy. METHODS: A retrospective study of 20 children with FBPase deficiency was conducted, based on their medical records. RESULTS: Six of the 20 children were given glycerol solution for the presence or possibility of brain edema during acute metabolic decompensation of hypoglycemia and metabolic acidosis; two of the six patients administered with glycerol were given dialysis. In four patients treated with glycerol alone without dialysis, two had no brain edema before glycerol administration but it developed later after the administration. These four patients treated with glycerol alone died or developed severe neurological complications. Fourteen patients who were not treated with glycerol solution had no brain edema and showed good prognosis. CONCLUSIONS: Glycerol solution, which contains fructose in Asian countries including Japan, should not be used as an osmotic agent for treatment of brain edema in patients who have hypoglycemia and retention-type metabolic acidosis, until FBPase deficiency is ruled out by measuring blood concentration of lactate. PMID- 12654062 TI - Screening for glucose-6-phosphate dehydrogenase deficiency using a modified formazan method: a pilot study on Filipino male newborns. AB - BACKGROUND: Glucose-6-phosphate dehydrogenase (G6PD) deficiency has increased prevalence rates in tropical Africa, tropical and subtropical Asia and some parts of the Mediterranean. Earlier studies on G6PD deficiency in the Philippines have shown prevalence rates of 4.5% to 25.7%. METHODS: In the present study, 3278 male newborns were screened for G6PD deficiency using the modified formazan method, a simple screening procedure affordable in the setting of a developing country. Subjects with positive screening results were recalled for confirmatory testing using a commercial assay kit for quantitative enzyme determination. RESULTS: Of the 3278 boys studied, 186 revealed positive screening results. Of the 186, 65 boys had confirmatory testing. Of these 65 boys, 45 were confirmed to have G6PD deficiency and 20 had normal results. This study reveals an incidence of G6PD deficiency of 3.9% among male Filipinos. CONCLUSIONS: This study recommends the inclusion of G6PD deficiency in the panel of disorders for newborn screening among Filipino newborns. PMID- 12654063 TI - Biochemical and molecular characterization of mutant hexosaminidase A in a Turkish family. AB - BACKGROUND: Tay-Sachs disease is a form of monosialoganglioside triaose (GM2) gangliosidosis that results from the mutations in the alpha-subunit gene of hexosaminidase A. In the B1 variant, the active site of the alpha-subunit of the enzyme is thought to be affected. In the present study, a patient who had previously been diagnosed as a B1 variant is further analyzed. The patient's parents and brother were also analyzed. METHODS: Single-stranded conformational polymorphism (SSCP) and DNA sequencing analysis were conducted in all cases. In addition, hexosaminidase A (Hex A) was isolated from leukocyte homogenates of the patient's parents and brother using DE 52 ion-exchange chromatography, and thermostability analyses of the isolated enzymes were performed. RESULTS: Hexosaminidase A of the parents was found to be more thermostable than normal Hex A. DNA sequencing analysis revealed a 12-bp deletion mutation in exon 10 of the Hex A gene. The patient was a homozygote and the parents were heterozygotes for the mutation, which could also be observed at the DNA double strands by SSCP analysis. These deleted bases are located within the catalytic domain of the alpha-subunit. CONCLUSIONS: The 12-bp deletion mutation in exon 10 of Hex A is responsible for the increased thermostability of the enzyme. Considering this mutation has previously been found in a Turkish Tay-Sachs patient, the patient in the present study may have another mutation on the Hex B gene that causes decreased thermostability of the enzyme. Thermal inactivation assay may not be sufficient for a correct diagnosis in such unusual cases. PMID- 12654064 TI - Hypophosphatemia in juvenile patients with systemic lupus erythematosus. AB - BACKGROUND: Hypophosphatemia is observed in both genetic diseases and acquired disorders, although it has never been described in systemic lupus erythematosus (SLE). The present study reports hypophosphatemia observed in patients with SLE, and investigates whether serum phosphorus (P) concentration is associated with disease activity and some cytokines. METHODS: Six girls with SLE (age 10 to 16 years) and seven age-matched controls were studied. Relationships between serum P and parameters that represented disease activity were evaluated. Interleukin (IL) 6 and tumor necrosis factor alpha (TNF-alpha) levels were measured by enzyme linked immunosorbent assay (ELISA). RESULTS: Two of the patients with SLE developed marked hypophosphatemia during active disease. Their serum P significantly correlated positively with serum complements, red blood cell (RBC) and platelet counts, and negatively with anti-double-stranded DNA antibody (dsDNAab). Three of the remaining four patients showed the same trend, in which their serum P significantly correlated with RBC, platelets, dsDNAab and/or serum complement concentrations. Serum and urinary IL-6 and serum TNF-alpha were higher in active SLE patients than those with inactive disease or in the control group. In addition, the concentrations of these cytokines significantly correlated inversely with serum P. Renal tubular reabsorption of P was significantly lower in patients with active disease both with and without lupus nephritis. CONCLUSIONS: Hypophosphatemia may be another sign of juvenile SLE, and serum P may represent disease activity. Both TNF-alpha and IL-6 may be related to hypophosphatemia in patients with SLE. Waste of P from renal tubules may be a possible mechanism of hypophosphatemia in SLE. PMID- 12654066 TI - Endothelin-1 in children with acute poststreptococcal glomerulonephritis and hypertension. AB - BACKGROUND: Endothelin-1 (ET-1), the most potent vasoconstrictor peptide, is known to play a role in arterial hypertension. In patients with acute poststreptococcal glomerulonephritis (APSGN) an increase in the production of ET 1 is suspected due to damaged endothelium, platelet activation and increased thrombin production in the glomeruli. The aim of the present study was to investigate whether the levels of plasma ET-1 are elevated in children with APSGN. Furthermore, we examined the association between plasma ET-1 levels and blood pressure levels in the same children. METHODS: We studied 18 children (14 boys) with APSGN (mean age 7.44 to approximately 2.82 years). Fourteen healthy children served as controls. The following parameters were evaluated: plasma ET 1, plasma atrial natriuretic peptide (ANP), plasma renin (Rn), serum aldosterone (Aldo), creatinine clearance (Ccr) and fractional excretion of sodium (FENa). RESULTS: The mean plasma ET-1 concentrations were higher in patients with APSGN (3.39 to approximately 1.86 pg/mL) compared to controls (1.40 to approximately 0.15 pg/mL; P=0.0001). Patients with APSGN also had higher plasma ANP concentrations (41.67 to approximately 27.99 pg/mL) than the controls (22.80 to approximately 4.24 pg/mL; P=0.011). Plasma Rn concentrations were lower in patients (24.54 to approximately 16.34 microU/mL) compared to controls (56.76 to approximately 32.36 microU/mL; P=0.027). A positive correlation was found between ET-1 plasma concentrations and the height of systolic or diastolic blood pressure (r=0.57, P=0.013 and r=0.53, P=0.023, respectively). CONCLUSIONS: Our results suggest that increased plasma ET-1 concentrations may play an important role in the pathogenesis of hypertension in children with acute poststreptococcal glomerulonephritis. PMID- 12654065 TI - Therapeutic effect of spiramycin in brucellosis. AB - OBJECTIVE: This study was undertaken to investigate the usefulness of spiramycin in treatment for brucellosis in an animal model. METHODS: Eighty-four Sprague Dawley rats were infected by intraperitoneal injection of Brucella melitensis suspension. Seven days after inoculation, four rats were selected randomly, killed and spleen cultures and Brucella standard tube agglutination test were carried out. All four rats were found to be infected. Eighty adult rats were randomly divided into four groups of 20 rats each. Tap water was given to the first group. Rifampicin 50 mg/kg per day and doxycycline 40 mg/kg per day were given to the second group, spiramycin 50 mg/kg per day orally was given to the third group, and a combination of spiramycin and rifampicin at the same dose and period was given to the fourth group. Duration of therapy regimens in all groups was 21 days. The spleens of all 80 rats were removed aseptically, homogenized, and placed onto Brucella agar plates to determine if viable bacteria were present. RESULTS: Bacterial growth occurred in all of the rats' spleens in the first group and in two rats' spleens in the spiramycin group. Mean colony forming unit (c.f.u.) values were at the highest in the first group. The effectivities of spiramycin and rifampicin-spiramycin were similar to rifampicin-doxycycline. There were no differences in the treatment results between the three groups that received combined rifampicin-doxycycline, rifampicin-spiramycin and only spiramycin (P>0.05). CONCLUSIONS: The results show that spiramycin cures experimental rat brucellosis and may be an effective alternative in the therapy of human brucellosis. PMID- 12654067 TI - Influence of heart surgery on magnesium concentrations in pediatric patients. AB - BACKGROUND: There have been only a few investigations into the effects of cardiopulmonary bypass (CPB) on the magnesium (Mg) concentrations in pediatric patients. The purpose of the present study was to compare the postoperative Mg concentrations and their recovery time to pre-surgical values in pediatric patients undergoing CPB for surgical repair of congenital heart disease. The incidence of dysrhythmia was also determined. METHODS: Twenty-seven pediatric patients undergoing open-heart surgery with CPB were enrolled in this study. Controls were 23 pediatric patients undergoing palliative surgery without CPB. Serum Mg (SMg) concentrations and ionized Mg (iMg) concentrations were measured at four sample points: 24 h before the surgery, immediately after the surgery, 24 h after the surgery and 48 h after the surgery. RESULTS: Serum Mg and iMg concentrations were significantly decreased after open-heart surgery. Immediately after the surgery, the mean SMg concentration was 64.1% and the mean iMg concentration was 68.8% of the pre-surgical values. The concentration of iMg showed quicker recovery than that of SMg, and returned to normal range 48 h after surgery. However, SMg and iMg concentrations after palliative surgery did not show significant differences from the pre-surgical values. The incidence of dysrhythmia was almost the same between the two groups; dysrhythmia was observed in two patients in the open-heart surgery group and in three patients in the palliative surgery group. CONCLUSIONS: Mg concentrations showed significant decrease after CPB; however, they showed quick recovery and did not increase the incidence of dysrhythmia in pediatric patients. PMID- 12654068 TI - Transcatheter occlusion of patent ductus arteriosus using tornado platinum coils. AB - BACKGROUND: The purpose of the present study was to develop a method to occlude a patent ductus arteriosus (PDA) using a tornado platinum coil, which is compatible with magnetic fields. METHODS: Twelve patients with a PDA (5 boys and 7 girls; 0.6 to 7 yrs; 6.5 to 22.1 kg) were enrolled. The minimum size of the PDA ranged from 0.2 mm to 3.6 mm. Either the anterograde or retrograde method was applied using a retrievable system that consisted of a 5-Fr multipurpose catheter and a 3 Fr bioptome. Three to 3.5 loops of the larger end of a tornado platinum coil were placed in the aortic ampulla and the remaining 0.5-1.0 loop of the smaller end of the coil was placed in the main pulmonary artery. RESULTS: In 11 patients with minimum size of PDA 0.05). CONCLUSIONS: The results of the present study showed that sensory integration, vestibular stimulation and neurodevelopmental therapy were effective in children with Down syndrome. It was concluded that when designing rehabilitation programs for children with Down syndrome, all treatment methods should be applied in combination, and should support each other according to the individual needs of the child. PMID- 12654074 TI - Pharmacokinetics of etoposide with intravenous drug administration in children and adolescents. AB - BACKGROUND: Pharmacokinetics of etoposide in Japanese children and adolescents has not been investigated. The objectives of the present study were (i) to document the pharmacokinetics of etoposide in Japanese children; (ii) to determine the intra- and interpatient variability in systemic etoposide exposure and (iii) to obtain insights into the age-pharmacokinetic parameter relationship. METHODS: Pharmacokinetic studies of etoposide, given at doses of 60-200 mg/m2 by intravenous (i.v.) route of administration, were conducted in 18 children and adolescents (aged <19 years) with malignant diseases. High performance liquid chromatography was used to measure the blood etoposide levels. RESULTS: Pharmacokinetic parameters (mean~SD) of the 14 patients (24 courses) who received etoposide 100 mg/m2 were as follows: peak serum concentration (Cmax), 18.5~6.4 microg/mL; trough serum concentration, 0.2~0.1 microg/mL; biological half-life (T1/2), 3.6~0.7 h; volume of distribution (Vd), 6.3~3.4 L/m2; area under the etoposide serum concentration-time curve (AUC), 129~38 hr x microg/mL; systemic clearance, 21.1~10.8 mL/min per m2. The T1/2, Vd, and AUC were not associated with age. An increase in etoposide dose per body surface area (BSA) was associated with increase in its Cmax and area under the time-concentration curve (AUC). Wide interpatient variability in these parameters was demonstrated. CONCLUSIONS: The present study demonstrated that: (i) Pharmacokinetics of etoposide in Japanese children and adolescents were similar to those in Caucasians. (ii) Increased exposure to etoposide was associated with the Cmax. A clear correlation between Cmax and AUC was also found. (iii) Selecting the dose of etoposide according to body surface area (BSA) might give an acceptable range of exposure for children more than 1 year of age. PMID- 12654075 TI - Factors associated with practice of procedures in pediatric-related areas among certified pediatricians. AB - BACKGROUND: Medical care in pediatric-related areas in which physicians other than pediatricians, such as surgeons, otolaryngologists, ophthalmologists, orthopedists and dermatologists, can also participate is one of the fields of pediatric primary care. Independent factors associated with the practice of specific procedures in pediatric-related areas by certified pediatricians in Japan were evaluated. METHODS: Data were collected from a nationwide survey of 1116 pediatricians who were selected by systematic sampling from the list of certificated members of the Japan Pediatric Society. The survey included questions relating to full- or part-time practice, types of practice setting, practice with or without pediatric subspecialty interest, numbers of procedures performed among 15 procedures in pediatric-related areas, and attitude toward practice of those procedures. RESULTS: Of 79% of respondents, 73% were analyzed (n=810). Among 15 procedures in pediatric-related areas, seven procedures performed by more than 30% of certificated pediatricians were evaluated. In comparison with certified pediatricians who annually performed three or less procedures (n=410), certified pediatricians who annually performed four or more procedures (n=400) were more likely to be young and male, working full-time and in a general hospital setting, practicing with a pediatric subspecialty interest. In multiple logistic regression analyses, young age (odds ratio [OR]: 0.8 per 10 years of age; 95% CI: 0.6-0.9), positive attitude toward practice (OR: 1.7; 95% CI: 1.6-1.9), male (OR: 1.7; 95% CI: 1.1-2.5), full-time practice (OR: 3.2; 95% CI: 1.5-6.9), and practice in general hospital setting (OR: 1.7; 95% CI: 1.2-2.4) were independently associated with more extensive practice of procedures in pediatric-related areas. CONCLUSIONS: A positive attitude toward medical care in pediatric-related areas is one of the most important factors associated with practice of procedures in these areas. An educational strategy for pediatricians to keep their motivation high for improved medical care in these areas should be planned in Japan. PMID- 12654076 TI - Prevention of antibiotic-associated diarrhea in children by Clostridium butyricum MIYAIRI. AB - BACKGROUND: Clostridium butyricum MIYAIRI (CBM) is a probiotic bacteria used for anti-diarrheal medicine in Japan. The preventive effect of CBM was investigated for antibiotic-associated diarrhea (AAD) in children. METHODS: One hundred and ten children who suffered from upper respiratory tract infection or gastroenteritis were divided into three groups. Twenty-seven of the patients received only antibiotics, 38 received CBM from the mid point of the antibiotic treatment and 45 concomitantly received CBM from the beginning of the antibiotic treatment. To examine the effects of CBM on AAD, the changes in intestinal flora were investigated. RESULTS: Diarrhea was observed in 59% of the subjects who received only antibiotics, and total fecal anaerobes, especially Bifidobacterium, were remarkably decreased. In contrast, diarrhea in the subjects who received CBM from either the middle or the beginning of the antibiotic therapy was decreased to 5% and 9%, respectively. Concomitant administration of CBM increased anaerobes and prevented the decrease of Bifidobacterium in the subjects who received antibiotics. CONCLUSIONS: Clostridium butyricum MIYAIRI is effective for both the treatment and the prophylaxis of AAD in children, as it normalizes the intestinal flora disturbed by antibiotics. PMID- 12654077 TI - Scheie syndrome (MPS-IS) presented as bilateral trigger thumb. PMID- 12654078 TI - Intraventricular hemorrhage in a full-term neonate associated with sinus venous thrombosis and homozygosity for the plasminogen activator inhibitor-1 4G/4G polymorphism. PMID- 12654079 TI - Duplication (22)(q11.22-q11.23) without coloboma and cleft lip or palate. PMID- 12654080 TI - Scleredema adultorum due to streptococcal infection. PMID- 12654081 TI - Aortopexy and bronchopexy for the management of severe tracheomalacia and bronchomalacia. PMID- 12654082 TI - Effective use of corticosteroid in a child with life-threatening plastic bronchitis after Fontan operation. PMID- 12654083 TI - Severely damaged aortic valve and cardiogenic shock in an infant with Kawasaki disease. PMID- 12654084 TI - Henoch-Schonlein purpura associated with hepatitis A infection. PMID- 12654085 TI - Case of shaken baby syndrome in Japan caused by shaking alone. PMID- 12654086 TI - C-reactive protein-mediated phagocytosis of Leishmania donovani promastigotes does not alter parasite survival or macrophage responses. AB - C-reactive protein (CRP) is an acute phase protein that binds to surface structures of a number of different organisms. Leishmania donovani express CRP ligand when first entering the mammalian host and CRP has been shown to alter macrophage function. The aim of this study was to investigate the functional significance of CRP-mediated uptake of L. donovani on survival of the parasite within human macrophages and macrophage cell responses to the infection. CRP opsonized L. donovani uptake was inhibitable by including excess CRP in the fluid phase, suggesting Fc receptor usage rather than indirect complement-mediated uptake. Comparing equivalent initial infection loads, parasite survival over 72 h within peripheral blood derived macrophages (PBMs) and differentiated U937 cells was unaltered by CRP. Whereas CRP increased macrophage responses to phosphorylcholine coated erythrocytes, no significant alteration in tumour necrosis factor-alpha, interleukin (IL)-10 or IL-12 production from PBMs was observed between CRP opsonized or unopsonized L. donovani promastigotes. Thus, in contrast to other systems, where CRP opsonization results in macrophage activation, Leishmania can use CRP to improve infection without inducing detrimental macrophage activation. PMID- 12654088 TI - Immunomodulatory properties of Ascaris suum glycosphingolipids - phosphorylcholine and non-phosphorylcholine-dependent effects. AB - Immunomodulatory properties of phosphorylcholine (PC)-containing glycosphingolipids from Ascaris suum were investigated utilizing immune cells from BALB/c mice. Proliferation of splenic B cells induced either via F(ab')2 fragments of anti-murine Ig (anti-Ig) or LPS was significantly reduced when the glycosphingolipids were present in the culture medium. However whereas the LPS mediated effect was dependent on the PC moiety of the glycosphingolipids, the result generated when using anti-Ig was not. Analysis of cell cycle status and mitochondrial potential indicated that the combination of the glycosphingolipids and anti-Ig reduced B cell proliferation, at least in part, by inducing apoptosis. Consistent with the observed suppression of B cell activation/cell cycle progression, investigation of the effect of glycosphingolipid pre-exposure on mitogenic B cell signal transduction pathways activated by anti-Ig, revealed a PC-independent inhibitory effect on dual (thr/tyr) phosphorylation and activation of ErkMAPKinase. The glycosphingolipids were also investigated for their inhibitory effect on LPS/IFN-gamma induced Th1/pro-inflammatory cytokine production by peritoneal macrophages. It was found that IL-12 p40 production was inhibited and in an apparently PC-dependent manner. Overall these data indicate that PC-containing glycosphingolipids of A. suum appear to have at least two immunomodulatory constituents - PC and an as yet unknown component. PMID- 12654087 TI - Endemically exposed asymptomatic individuals show no increase in the specific Leishmania (Viannia) panamensis-Th1 immune response in comparison to patients with localized cutaneous leishmaniasis. AB - In Colombia, most cases of human cutaneous leishmaniasis are caused by Leishmania (Viannia) panamensis. Interestingly, up to 30% of the exposed population do not suffer from clinical leishmaniasis although it is likely that they are continuously infected with Leishmania parasites. Since it is believed that the induction of efficient Th1 immune responses protects against Leishmania infections both in humans and in animal models, we determined if endemically exposed asymptomatics showed stronger Leishmania-specific Th1 immune responses than patients with active localized cutaneous leishmaniasis (LCL). We found that Montenegro skin test responses were slightly higher among asymptomatic individuals compared to patients suffering from LCL. However, PBMC from patients with LCL showed similar Leishmania-specific proliferative responses compared to PBMC from asymptomatic individuals. Furthermore, PBMC from both groups also secreted similar amounts of IFN-gamma, IL-12p40 and IL-10 after in vitro exposure to L. panamensis. No IL-4 was detected in the supernatants. Taken together our results suggest that lack of LCL development in endemically exposed asymptomatics cannot be explained by stronger systemic anti-Leishmania Th1 immune responses or decreased Th2 responses in these individuals in comparison to individuals who develop LCL. It may be possible that other mechanisms are responsible for resistance to cutaneous leishmaniasis in Colombia in endemically exposed asymptomatics. PMID- 12654089 TI - Expression of inducible nitric oxide synthase (iNOS) mRNA in target organs of lethal and non-lethal strains of murine malaria. AB - Nitric oxide (NO) is a putative mediator of the immunological and/or pathological responses to malaria, consequently it is a potential target for novel drug therapy. Numerous cell types increase expression of inducible nitric oxide synthase (iNOS) under inflammatory conditions, the most relevant stimuli being cytokines and endotoxins. In this study the expression of iNOS mRNA in several target organs (brain, liver, spleen) of malaria have been investigated in MF1 mice during lethal Plasmodium (P.) berghei and non-lethal P. c. chabaudi infection. In P. berghei malaria, iNOS mRNA decreased in liver and was unchanged in spleen during the period of rising parasitaemia, but increased in both organs late in the infection, when parasitaemia was high and death imminent. In mice infected with P. c. chabaudi, spleen iNOS mRNA increased progressively throughout the early, peak and recovery periods of parasitaemia, but decreased in liver. Brain iNOS mRNA decreased in samples collected throughout the time courses of both infections. Hence it is evident that changes in iNOS mRNA in murine malaria depend upon the tissue, day of infection, degree of parasitaemia and strain of Plasmodium. These data indicate induction of iNOS mRNA in the spleen has a role in combating these strains of Plasmodium in MF1 mice. Failure to clear lethal P. berghei parasitaemia was associated with increased iNOS mRNA expression in the liver, which may contribute to the pathology of this malaria. PMID- 12654090 TI - Neutropenia augments experimentally induced Schistosoma japonicum egg granuloma formation in CBA mice, but not in C57BL/6 mice. AB - The present study was designed to investigate the role of neutrophils during the development of Schistosoma japonicum egg granulomas, in C57BL/6 and CBA mice. Laid eggs were implanted into the liver and monoclonal antibody, RB6-8C5, was used to eliminate neutrophils. After daily antibody treatment between days 9 and 13 of egg implantation, both strains of mice showed a marked decrease in neutrophil infiltration and coagulative hepatocyte necrosis at 2 weeks. At 4 weeks, after antibody administration every other day between days 16 and 26, granuloma formation in C57BL/6 mice was not affected by the treatment, whereas CBA mice exhibited a significant increase of reactions. Neutropenia augmented the Th2 cytokine response (IL-4, IL-13 and IL-5), but not for IFN-gamma at any time point examined and in either strain of mice. Higher levels of IL-4 and IL-13 were noted in CBA mice at early and late stages of granuloma formation, compared to C57BL/6 mice. There was also a striking difference in IL-13 production between the two strains. Our results indicate that neutropenia is associated with a significant augmentation of S. japonicum egg-induced granuloma formation in CBA mice, probably through increase in Th2 cytokines, however, the effects differ between early and late stages and between high and low responders. PMID- 12654091 TI - Plasma levels of interleukin-18 and interleukin-12 in Plasmodium falciparum malaria. AB - Interleukin (IL)-18 produced primarily by mononuclear phagocytes synergizes with IL-12 for interferon-gamma production from T, B and natural killer cells. It has been also demonstrated that, in Plasmodium falciparum malaria, IL-18 could have an immunoregulatory function. The aim of this study was to detect the plasma levels of IL-12 and IL-18, using an enzyme-linked immunosorbent assay, in 105 African children affected by mild and severe Plasmodium falciparum malaria to correlate the production of these cytokines with the severity of the disease. The levels of IL-18 and IL-12 were higher (25.7 +/- 7.6 pg/ml and 17.1 +/- 7.8 pg/ml, respectively) in children with mild malaria than in children with a severe form of the disease (21.5 +/- 10 pg/ml and 13.2 +/- 5.5 pg/ml, respectively). A positive correlation was observed between IL-18 and IL-12. This finding suggests that the production of these two cytokines (IL-18 and IL 12) may be coregulated and both have an immunoregulatory effect on the immune response in Plasmodium falciparum infection. PMID- 12654092 TI - P38 mitogen-activated protein kinase influence on the production of IL-10 in human schistosomiasis mansoni. AB - Schistosomiasis mansoni remains a significant public health problem in many parts of the tropics and subtropics. Clinical manifestations range from the asymptomatic intestinal form through to the hepatosplenic form of the disease, a potentially lethal clinical condition in a subsection of the exposed population. In this study, we investigated the mechanisms by which interleukin (IL)-10 production could be differentially controlled in patients with the intestinal and hepatosplenic forms of the disease, as IL-10 may play a fundamental role in the development of the hepatosplenic disease state. It is reported that p38 mitogen activated protein (MAP) kinase signalling, and in particular p38 MAPK activation, is central to IL-10 production of cells from patients with schistosomiasis. Furthermore, the difference in the levels of activated p38 MAPK and the activation transcription factor (ATF-2), may explain the difference in the amount of IL-10 produced by cells from intestinal and hepatosplenic patients. We suggest that the type of immune response triggered in patients with hepatosplenomegaly could be influenced by the levels of phosphorylated p38 MAPK. PMID- 12654093 TI - Activation of interleukin 1beta gene transcription by human cytomegalovirus: molecular mechanisms and relevance to periodontitis. AB - In recent years, studies have demonstrated an association between human cytomegalovirus (HCMV) and destructive periodontal disease. It has been shown that reactivation of HCMV in periodontitis lesions may be related to progressing periodontal disease. Several possible mechanisms by which HCMV exerts periodontopathic potential have been previously proposed. These are reviewed and include the upregulation of bone resorptive cytokines such as interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) by active HCMV infection at the periodontitis site. This review focuses on the molecular basis of IL-1beta gene activation by HCMV immediate early (IE) gene products. A novel hypothesis is also described whereby HCMV plays a significant role in the pathogenesis of periodontal disease by the ability of its IE proteins to strongly transactivate IL-1beta gene expression. More studies are needed to further explore this hypothesis and clarify the association between HCMV and periodontitis. PMID- 12654094 TI - Immunoglobulins in saliva of preterm and full-term infants. AB - The aim of this longitudinal study was to determine salivary levels of total IgA, IgG and IgM in 84 preterm and 214 full-term infants, from birth to 18 months of age. Unstimulated whole saliva was collected from each infant at birth, and subsequently at 3-monthly intervals. Immunoglobulin levels were estimated using an ELISA technique. At birth, IgA was detected in 147/214 (69%) full-term infants but only 47/84 (56%) preterm infants (P < 0.01). In the case of IgG, 61% of full term and 56% of preterm infants showed detectable levels, whereas IgM was found in 71% and 73%, respectively. Levels of IgA and IgG rose from birth to 18 months, whereas IgM levels did not change significantly. Increases in salivary levels of IgA were associated with introduction of solid foods (P < 0.001), as well as tooth eruption (P < 0.001). Our results indicate that the majority of full-term and preterm infants are orally immunocompetent at birth. PMID- 12654095 TI - Helicobacter pylori and Campylobacter rectus share a common antigen. AB - AIM: The aim of this study was to investigate the presence of antigens with immunological cross-reactivity in periodontopathogenic bacteria and Helicobacter pylori, the pathogen associated with gastritis and peptic ulcers in human. MATERIALS AND METHODS/RESULTS: Among the putative periodontopathogens tested (Actinobacillus actinomycetemcomitans, Campylobacter rectus, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia and Treponema denticola), cross-reactive bands were only detected in C. rectus by SDS PAGE/Western immunoblotting analysis using a polyclonal antibody directed to H. pylori cells. One of these cross-reactive antigens, a 64-kDa band antigen, also reacted with a monoclonal antibody directed to the human heat shock protein (HSP) 60. The N-terminal amino acid sequence of this C. rectus protein revealed a high degree of homology with corresponding regions of other HSPs belonging to the HSP60 family, indicating that the 64-kDa antigen was a GroEL protein. The nucleotide sequence of the C. rectus GroEL protein coded for a 547 amino acid protein with a predicted size of 57.8 kDa. Comparison of the alignment of the deduced amino acid sequence of the GroEL protein of C. rectus with that of H. pylori showed a high degree of similarity throughout its length (76.8%). GroEL protein from C. rectus possessed the ability to stimulate production of IL-6 by a confluent monolayer of human gingival epithelial cells and was cytotoxic when used at a high concentration. CONCLUSIONS: This study reveals an immunological relationship between H. pylori and C. rectus, and clearly indicates that one of the shared antigens is a GroEL protein possessing a biological activity that might play a role in the initiation and progression of periodontal disease. PMID- 12654097 TI - Fusobacterium nucleatum impairs serum binding to Porphyromonas gingivalis biofilm. AB - Mouse immune sera obtained by immunization with Fusobacterium nucleatum and then Porphyromonas gingivalis demonstrated an impaired binding capacity to P. gingivalis-biofilm and lower avidity to P. gingivalis when compared with sera obtained from mice immunized with P. gingivalis alone. PMID- 12654096 TI - Influence of periodontal disease on Th1/Th2-type cytokines in saliva of HIV positive individuals. AB - Cytokines present during immune responses have a tremendous influence on resistance/susceptibility to oral diseases including periodontal disease and oral opportunistic infections in the immunocompromised individual, as seen by altered Th cytokines in saliva with human immunodeficiency virus (HIV) disease progression and oropharyngeal candidiasis. This study was designed to evaluate whether the presence of severe periodontal disease has any influence on Th cytokines in saliva of HIV-positive persons. For this, saliva from a cohort of HIV-positive persons with mild or severe periodontitis was evaluated for Th cytokines. A dominant Th2-type cytokine profile in saliva was validated in HIV positive subjects with considerable immune suppression, irrespective of periodontal disease status. However, no significant differences in concentrations of Th1- or Th2-type cytokines in saliva were observed when stratified by periodontal status. Thus, the lack of salivary influences by periodontitis eliminates periodontal disease as a variable in interpretations regarding correlates of local cytokines during oral manifestations of HIV. PMID- 12654098 TI - Defensin-induced adaptive immunity in mice and its potential in preventing periodontal disease. AB - The severity of periodontal disease is dependent on a combination of host, microbial agent and environmental factors. One strong correlate related to periodontal disease pathogenesis is the immune status of the host. Here we show that human neutrophil peptide (HNP) defensins or human beta-defensins (HBD), co administered intranasally with the antigen ovalbumin (OVA), induce unique immune responses that if used with microbial antigens may have the potential to hinder the pathogenesis of periodontal disease. C57BL/6 mice were immunized intranasally with phosphate buffered saline (PBS) containing 1 micro g HNP-1, HNP-2, HBD1 or HBD2 with and without 50 microg OVA. At 21 days, isotypes and subclasses of OVA specific antibodies were determined in saliva, serum, nasal wash, bronchoalveolar lavage fluid, and fecal extracts. OVA-stimulated splenic lymphoid cell cultures from immunized mice were assessed for interferon (IFN)-gamma, Interleukin (IL)-4 and IL-10. In comparison with mice immunized with only OVA, HNP-1 and HBD2 induced significantly higher (P < 0.05) OVA-specific serum IgG, lower, but not significant, serum IgM and significantly lower (P < 0.05) IFN-gamma. In contrast, HNP-2 induced low OVA-specific serum IgG and higher, but not significant, serum IgM. HBD1 induced significantly higher (P < 0.05) OVA-specific serum IgG, higher, but not significant, serum IgM, and significantly higher (P < 0.05) IL-10. The elevated serum IgG subclasses contained IgG1 and IgG2b. PMID- 12654099 TI - Evaluation of root canal microorganisms isolated from teeth with endodontic failure and their antimicrobial susceptibility. AB - Studies of the microbiota from the canals of teeth with failure of endodontic therapy have revealed that it differs markedly from that of untreated necrotic dental pulps. This study aimed to evaluate the microbiota of 30 root-filled teeth with persisting periapical lesions and to test the antibiotic susceptibility of the most prevalent species. Microbial samples, isolation and speciation were done using advanced microbiologic techniques for anaerobic species. A total of 55 bacterial species were isolated, 80% were gram-positives and 58% facultative anaerobic microorganisms. The bacterial genera most frequently recovered were Enterococcus, Streptococcus, Peptostreptococcus and Actinomyces. Antibiotic sensitivity of Enterococcus faecalis and Peptostreptococcus spp. was accomplished with the E-test system. All species studied were susceptible to benzylpenicillin, amoxicillin, amoxicillin combined with clavulanate. However, 20% of the E.faecalis strains were resistant to erythromycin and 60% to azithromycin. It was concluded that microbial flora in canals after endodontic failure comprised predominantly facultative anaerobes and gram-positive species. E.faecalis was the species most frequently isolated and showed erythromycin and azithromycin resistance among the isolates. PMID- 12654100 TI - Cytomegalovirus and Epstein-Barr virus DNA transcription in endodontic symptomatic lesions. AB - OBJECTIVES: Productive Herpesviridae infections are implicated in the etio pathogenesis of aggressive periodontitis. However, virtually nothing is known about a possible role of herpesviruses in pulpal and periapical pathosis. This study employed a cDNA analysis to determine transcription of human cytomegalovirus (HCMV), Epstein-Barr virus (EBV) and herpes simplex virus (HSV) in 14 recalcitrant periapical lesions and in 2 periapical healthy control sites. METHODS: Periapical samples were collected in conjunction with periapical surgery and kept frozen until virologic examination. RNA was isolated from periapical tissue by using a guanidinium isothiocyanate-acid phenol procedure (TRIZOL LS Reagent, GIBCO BRL, Rockville, MD). cDNAs were amplified by means of oligonucleotides targeting highly conserved regions of the test viruses and the RT-PCR-100 amplification kit (Sigma-Aldrich, St Louis, MO). Standardization of PCR primer sensitivity and validation was carried out according to established methods. Amplification products were identified by agarose gel electrophoresis. RESULTS: HCMV transcript was detected in 12 of 13 symptomatic and in 1 asymptomatic periapical lesion. EBV transcript was demonstrated in 8 of the 13 symptomatic lesions but not in the asymptomatic periapical lesion. HCMV and EBV dual transcription occurred at higher frequency in periapical lesions showing radiographic bone destruction of 5 mm x 7 mm or larger than in smaller size lesions (P = 0.03; Chi-squared test). No HCMV or EBV transcription was identified in the 2 healthy control sites. HSV transcript was not detected in any study site. CONCLUSION: The present data suggest that HCMV or EBV infections participate in the pathogenesis of periapical symptomatic lesions. Herpesviruses may produce periapical pathosis as a direct result of viral infection and replication, or as a consequence of virally induced impairment of the host defense and subsequent increased virulence of resident bacterial pathogens. PMID- 12654101 TI - Acid-neutralizing activity during amino acid fermentation by Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum. AB - Acid-neutralizing activity during amino acid fermentation by washed cells of Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum was studied. When the washed cells of these strains were anaerobically incubated in the presence of aspartylaspartic acid or glutamylglutamic acid for P. gingivalis, aspartic acid for P. intermedia and glutamic acid for F. nucleatum at an initial pH of 5.0 or 5.5, the pH of the incubation mixtures rose toward neutral. F. nucleatum had the highest acid-neutralizing activity, followed by P. intermedia and P. gingivalis. The P. intermedia and F. nucleatum cells were used to measure the amounts of base produced at a fixed pH of 5.0. These cells generated significant amounts of base at pH 5.0 along with the production of organic acids and ammonia from aspartic or glutamic acid. Acid-base balance theoretically calculated from the amounts of consumed substrate and end products implies that the acid-neutralizing activity was derived from the decrease in acidity during the fermentation of amino acid into organic acids and ammonia. PMID- 12654102 TI - Prevalence of Csh-like fibrillar surface proteins among mitis group oral streptococci. AB - The prevalence of Csh-like fibrillar surface proteins among oral streptococci was investigated by ELISA and by immunoelectron microscopy using antiserum raised to recombinant fragments of CshA of Streptococcus gordonii DL1. The majority of S. gordonii, Streptococcus sanguis and Streptococcus oralis strains tested elaborated short (ca. 50-80 nm long) surface fibrils and reacted with antiserum to the amino acid repeat region of CshA, demonstrating the widespread nature of Csh-like proteins among these species. In contrast, reactivity with antiserum raised to the adhesion-mediating non-repetitive region of CshA was more restricted. On the basis of the ELISA results, several isolates were selected for immunogold analysis using CshA antisera. Immunogold-negative staining showed a surface distribution of 10 nm gold particles consistent with antibody binding to short fibrils. Long fibrils (>150 nm long), where present, were not significantly labelled with gold. The results suggest that some of the short peritrichous fibrils on many mitis group streptococci comprise Csh-like fibrillar protein. Further, the data are consistent with our hypothesis that the antigenically conserved amino acid repeat region of Csh-like proteins forms a scaffold for cell distal presentation of the amino-terminal non-repetitive region that, at least in S. gordonii DL1, functions as an adhesin. PMID- 12654103 TI - Influence of Enterococcus faecalis proteases and the collagen-binding protein, Ace, on adhesion to dentin. AB - Enterococcus faecalis is a pathogen that persists in medicated root canals. Here, we tested the hypothesis that the E. faecalis proteases, serine protease and gelatinase, and the collagen-binding protein (Ace) contribute to adhesion to the root canal. Scanning electron microscopy was used to examine dentin binding by four E. faecalis strains: OG1RF, the wild type, and three mutant derivatives of OG1RF, TX5128, TX5243 and TX5256 deficient in serine protease and gelatinase, serine protease, and Ace, respectively. For each strain, 20 root halves were exposed to 3 x 10(9) to 5 x 10(9) cells/ml for 6 h, and 50 fields per root half were examined for adherent bacteria. Statistical analysis revealed that adherence of OG1RF was significantly greater than the mutant strains (P < 0.001), while significant differences were not detected between the protease mutants. The data indicate that serine protease and Ace aid E. faecalis binding to dentin, while the role of gelatinase is uncertain. PMID- 12654105 TI - Concurrent oral shedding of feline calicivirus and feline herpesvirus 1 in cats with chronic gingivostomatitis. AB - Oral mucosal salivary samples were collected from 25 cats with chronic gingivostomatitis and 24 cats with periodontal disease. Viral culture and isolation of feline calicivirus and feline herpesvirus 1 were performed. Eighty eight per cent of cats with chronic gingivostomatitis were shedding both viruses, compared to 21% of cats without chronic oral inflammatory disease. Cats with chronic gingivostomatitis are significantly more likely to concurrently shed both feline calicivirus and feline herpesvirus 1 than are cats with classical periodontal disease. PMID- 12654104 TI - Amended biochemical characteristics and phylogenetic position of Treponema medium. AB - Umemoto et al. (1997, Int J Syst Bacteriol 47, pp. 67-72) proposed spirochete strain G7201, isolated from the periodontal pocket of an adult patient, as a new species, Treponema medium. They deposited this strain in the American Type Culture Collection (ATCC) as type strain ATCC 700293T. Recently, ATCC suggested that there is a discrepancy between the previous report and the results obtained by ATCC in biochemical tests on T. medium ATCC 700293T. In this study, we re examined and verified the biochemical characteristics of T. medium. The fermentation pattern of carbohydrates of T. medium resembled that of Treponema vincentii and Treponema denticola, but T. medium was clearly differentiated from T. vincentii in the production of indole, and from T. denticola in the hydrolysis of esculin. Also, sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS PAGE) protein profile analysis and phylogenetic comparison of 16S rDNA sequences revealed that T. medium is clearly differentiated from any established treponemal species, which supports the validity of the proposal of Treponema medium as a new species. PMID- 12654107 TI - Teaching about professionalism. PMID- 12654109 TI - Equal opportunity does not produce equity: (not) getting into medical school. PMID- 12654110 TI - Spoiled for choice. PMID- 12654111 TI - Whistle blowing. PMID- 12654112 TI - The Physick Garden: an interview with R M Harden. PMID- 12654113 TI - Does exposure to medically underserved areas during training influence eventual choice of practice location? AB - INTRODUCTION: There are an increasing number of communities within the United States that have limited or no access to primary healthcare. In recognition, many medical schools now provide opportunities and activities that offer exposure to these demographic areas in order to increase the presence of and community access to medical care and to promote these locations as practice site choices for graduating students. Evaluation of these enhancements has led to doubts whether this exposure timing is optimal in promoting practice in these settings. The purpose of this study is to identify whether early exposure(s) to medically underserved settings prior to medical school is associated with eventual choice of practice location. METHODS: Utilising a cross-sectional design, 450 US Family Medicine residency programmes were surveyed. From these, 775 participants responded to a standardised self-administered questionnaire on indicators associated with medically underserved area (MUA) exposure. RESULTS: Early MUA exposures combined with medical training experiences in underserved settings have a positive effect on later practice site choice. Identification of these attributes may be useful in considering determinants that impact eventual choice of practice location. PMID- 12654114 TI - The West Midlands Matching Scheme 2000: a survey of participant satisfaction and outcomes. AB - CONTEXT: The West Midlands Matching Scheme has been in operation since 1999 and is one of a number of schemes used for the allocation of pre-registration house officer positions in the UK. METHOD AND RESULTS: A questionnaire was distributed amongst candidates applying to the scheme in 2000. A response rate of 73.7% was obtained (160 replies from a sample population of 217). Half the responders reported to be dissatisfied with the explanation of the matching scheme given. A total of 66.6% were satisfied with the scheme being run via the Internet and E mail. For 6-month positions in medicine, 85% of the candidates were happy with the job they had received. The figures for surgical jobs were similar and for rotations even higher. However, 23.8% of the sample was missing one or two jobs at the end of the matching process. This figure was still high when non responders were taken into account. This group was not happy with the way in which the remaining jobs were distributed to them (71% dissatisfied). CONCLUSION: Although the matching scheme matched a high proportion of responders to jobs they were satisfied with, there was a significant group without jobs who were unhappy with the scheme. Some of those with jobs were also dissatisfied with the jobs they had received. Both problems should be addressed in future schemes. Further work in this area should examine the characteristics of the group without jobs and look at the success of other schemes in order to determine the optimal way in which to allocate PRHO jobs. PMID- 12654115 TI - Doctors' reasons for rejecting initial choices of specialties as long-term careers. AB - OBJECTIVES: To report on rejected choices of specialty as long-term careers and reasons for rejection. DESIGN: Postal questionnaire survey. SETTING: United Kingdom. PARTICIPANTS: All graduates of 1996 and 1999 from UK medical schools during their first year after qualification. MAIN OUTCOME MEASURES: Any career choice which had been seriously considered and rejected and the reason for its rejection. RESULTS: In all, 33.1% (1871) of respondents named a rejected choice and gave reasons for its rejection. Disproportionately high numbers rejected the surgical specialties, paediatrics and obstetrics and gynaecology (O&G), using the specialty distribution of positive choices as the comparator. Relatively few doctors rejected general practice (GP) after giving it serious consideration. Doctors rejecting the hospital medical and surgical specialties or paediatrics were most likely to specify reasons relating to quality of life. Three-quarters of the graduates of 1999 who rejected O&G did so because of poor career prospects. CONCLUSIONS: Quality of life issues, and concerns about working relationships, are sufficiently influential to persuade many doctors to abandon an initial choice of medical career. It is unlikely that much of the decline in entry to GP is attributable to rejection of GP by doctors who initially chose it. The decline must therefore represent an increase in the number of doctors who had never seriously considered it as a long-term career choice. PMID- 12654116 TI - Critical factors in career decision making for women medical graduates. AB - BACKGROUND: Within the next 30 years there will be equal numbers of women and men in the medical workforce. Indications are that women are increasing their participation in specialties other than general practice, although at a slower rate than their participation in the workforce as a whole. To inform those involved in training and employment of medical women, this study investigated the influencing factors in career decision making for female medical graduates. METHODS: A total of 305 women medical graduates from the University of Auckland responded to a mail survey (73% response rate) which examined influences on decision making, in both qualitative and quantitative ways, as part of a larger survey. RESULTS: Most women were satisfied with their careers. The principal component analysis of the influencing factors identified four distinct factors important in career choice - interest, flexibility, women friendliness and job security, although the first two of these were rated more highly than the others. CONCLUSIONS: Barriers to full participation by medical women in training and employment need to be systematically examined and removed. This is not only to allow women themselves to reach their full potential, but for workforce and socio economic reasons. Initiatives that allow and value more flexible training and work practices, particularly through the years of child raising, are necessary for women and the health care workforce at large. PMID- 12654117 TI - An evaluation of a national scheme for continuing professional development (CPD) for career grade doctors: the Royal College of Paediatrics and Child Health's programme for paediatricians evaluated by focus group methodology. AB - OBJECTIVES: To evaluate a national continuing professional development (CPD) scheme through the views and experiences of its participants. METHODS: A qualitative focus group methodology was used. Forty-nine career grade paediatricians were allocated to nine focus groups according to their work settings, ages and posts. All groups discussed the following: (a) ease or difficulty of achieving CPD requirements; (b) whether a personal professional development plan (PDP) enabled the planning process for CPD; (c) whether CPD participation facilitated changes in practice, and (d) their views on the CPD system in operation at the time of the study. RESULTS: Taking part in CPD had a positive effect on participants' clinical practice and broadened their professional and academic base. The main difficulties in achieving CPD were variable financial support (especially for non-consultant staff) and pressure due to extra work accumulating during study leave. The quality of internal CPD meetings was inconsistent. A professional development plan was useful for planning CPD needs, but there was a conflict between the educational and training needs of the individual and the employing NHS Trusts. The scheme's guidelines were clear but there were reservations about its layout and the system of recording points. CONCLUSION: This study enabled discovery of the views of paediatricians participating in a national CPD scheme operated by the Royal College of Paediatrics and Child Health. Consequently, changes were made to the scheme's presentation (but not content) and process of recording points. Barriers to achieving CPD requirements were identified and are being monitored. Efforts to involve non-consultant career grade paediatricians have been increased. PMID- 12654118 TI - Assessing students' personal and professional development using portfolios and interviews. AB - BACKGROUND AND PURPOSE: Medical schools are placing more emphasis on students' personal and professional development (PPD) and are seeking ways of assessing student progress towards meeting outcome goals in relation to professionalism. The Faculty of Medicine at the University of Sydney sought an assessment method that would demonstrate the value of reflection in attaining PPD, provide feedback and encourage students to take responsibility for setting and achieving high standards of performance. METHODS: The instruments used to assess Year 1 students in PPD are a portfolio and interview. This assessment format encourages students to explore ideas and values that are important to them and relevant to the PPD theme. A confidential interview, based on the PPD goals, is held with a faculty member who has read the student's portfolio. RESULTS: In 1997/98, 96% of students agreed that they had engaged in useful reflection on their approach to the course and 91% agreed that the experience was worthwhile. A further 76% of students agreed that they could see opportunities to modify their approach in some ways as result of this exercise. CONCLUSION: Sustained PPD is essential in equipping doctors for the varied stresses of careers in medicine. Despite, or perhaps because of, the latitude in the Year 1 assessment, both students and faculty members found the process of value. This form of assessment acknowledges that the most valid assessment formats cannot always be made reliable and that in some parts of the curriculum it is more important to demonstrate trust in students' own motivation to become competent and mindful practitioners. The fact that the portfolio and interview are the only summative assessments in the first year emphasises the importance that the Faculty places on PPD. PMID- 12654119 TI - Fostering students' personal and professional development in medicine: a new framework for PPD. AB - CONTEXT: Altruism, accountability, duty, integrity, respect for others and lifelong learning are qualities that have been identified as central to medical professionalism. However, we do not have a systematically developed understanding of what is needed to optimise medical students' personal and professional development (PPD). We need some level of agreement on how to teach and assess PPD, but traditional educational methods may not be strong determinants of students' or graduates' actual behaviour in clinical settings. AIMS: This paper considers the factors that demonstrably influence doctors' behaviour as a contribution to the development of a model for considering PPD within the broader context of medical practice. The model presented acknowledges that behaviour change comes about through a number of influences including education, feedback, rewards, penalties and participation. These elements can be plotted against the cognitive, affective and metacognitive processes that are intrinsic to learning. IMPLICATIONS: A framework that promotes the consideration of all of these factors in PPD can provide guidance for schools undergoing curriculum reform and inform further research into one of the most important and challenging aspects of medical education. PMID- 12654120 TI - To be and not to be: the paradox of the emerging professional stance. AB - PURPOSE: Understanding how students resolve professional conflict is essential to teaching and evaluating professionalism. The purpose of this study was to refine an existing coding structure of rationalizations of student behaviour, and to further our understanding of students' reasoning strategies in the face of perceived professional lapses. METHODS: Anonymous essays were collected from final year medical students at two universities. Each essay included a description of a specific professional lapse, and a consideration of how the lapse was dealt with. Essays were analysed using grounded theory. The resulting coding structure was applied using NVivo software. RESULTS: Twenty essays, containing 147 instances of articulated reasoning, were included. Three major categories (and several subcategories) of reasoning strategies emerged: Narrative Attitude (deflection or reflection), Dissociation (condescension or identity mobility), and Engagement (with associated action or no action). This data set revealed a wider range of Narrative Attitude than in the original study, confirmed the dominance of Dissociation as a reasoning strategy, and, perhaps paradoxically, also revealed significant evidence of action on the part of the students (predominantly directed towards dealing with the consequences of a lapse or confronting the lapser). Most of these actions were perceived to be ineffective. CONCLUSIONS: Encountering a professional lapse can be a paradoxical and profoundly disordering experience for students. When students report these experiences, they invoke reasoning strategies that enable them to re-story the lapse. Their methods of re-storying provide insight into the double-binds that students experience, their efforts to transcend these double-binds, and, through these, their emerging professional stance. PMID- 12654121 TI - CELT: a computerised evaluative learning tool for continuing professional development. AB - OBJECTIVES: To evaluate a computerised, evaluative learning tool (CELT) designed to encourage self-directed learning and help users make changes in practice following learning. The study aimed to evaluate how CELT was used and to ascertain user perceptions of the program. DESIGN: Qualitative analysis of interviews and quantitative analysis of entries made using the software. SETTING AND SUBJECTS: West of Scotland region, comprising six Health Board areas with a total of 2176 general practitioners (GPs), 39 of whom took part in the study. RESULTS: Of the 39 GPs who started on the project, 34 used CELT. Of these 34, 28 GPs sent in files and six did not. Of the 28 GPs who sent in files, 25 entered data and 76% (22/29) considered the program easy to use. The program was used 7 days a week during the day and night. It raised participants' awareness of the educational value of everyday experiences and led to increased thought about learning. In 41% (45/111) of entries there was evidence that some action had been initiated by users as a result of learning. CONCLUSIONS: CELT was designed to encourage self-directed learning and help users make changes in practice following learning. The study has shown that it can be used to deliver individual continuing professional development. It encourages a disciplined approach to learning, promotes thought about learning needs and increases the ability of GPs to learn from everyday experiences. In some instances, users were able to apply what had been learned. PMID- 12654123 TI - Experimental study design and grant writing in eight steps and 28 questions. AB - While writing a grant proposal may take a few days, the planning of the study takes much longer and requires thoughtful consideration. The use of a systematic and itemised approach can help in planning crucial details of a study. An eight step, 28-question, iterative approach is proposed to help with the careful planning of experiments in order to maximise the researchers' chances of acceptance when submitting the study for funding and its results for publication. The steps include defining a relevant research question; selecting instrumentation, study design and statistics; determining sample size and sampling procedure; ensuring data quality throughout data collection and analysis; setting personnel and budget requirements, and writing a convincing grant proposal. Reviewers pay particular attention to the importance of the research topic and question, the presence of a clear problem statement and up to date review of the literature, the use of an optimal design and instrumentation, a sufficient and unbiased sample, and appropriate and well applied statistics. They also appreciate a clear and easy to follow proposal. The research question is the keystone of the entire enterprise, followed by the selection of an optimal study design and the control of possible confounding variables. No study is perfect. The researchers must constantly weigh advantages and disadvantages and select the most scientifically sound and feasible alternatives. While the steps and questions presented are best applied to experimental studies, the principles are also applicable to a wide range of questions and observational, evaluative and qualitative designs. PMID- 12654122 TI - Students' attitudes and potential behaviour with regard to whistle blowing as they pass through a modern medical curriculum. AB - OBJECTIVE: To examine students' attitudes and potential behaviour with regard to whistle blowing as they progress through a modern undergraduate medical curriculum. DESIGN: Cohort design. SETTING: University of Glasgow Medical School. SUBJECTS: A cohort of students entering Glasgow University's new learner-centred, integrated medical curriculum in October 1996. METHODS: Students' pre- and post Year 1, post-Year 3 and post-Year 5 responses to the whistle blowing vignette of the Ethics in Health Care Instrument (EHCI) were examined quantitatively and qualitatively. Analysis of students' multichoice answers enabled measurement of movement towards professional consensus opinion. Analysis of written justifications helped determine whether their reasoning was consistent with professional consensus and enabled measurement of change in knowledge content and recognition of the values inherent in the vignette. Themes in students' reasoning behind their decisions of whether or not to whistle blow were also identified. RESULTS: There was little improvement in students' performance as they progressed through the curriculum in terms of their proposed behaviour on meeting the whistle blowing scenario. There was also no improvement in the quality of justifications provided. Students' reasoning on whether or not to whistle blow was found to change as the curriculum progressed. CONCLUSIONS: The EHCI has the potential to elicit students' attitudes towards ethical issues at entry to medical school and to measure change as they progress through the curriculum. Students should be encouraged to contemplate dilemmas from all ethical standpoints and consider relevant legal implications. Whistle blowing should be addressed as part of the wider domain of professionalism. PMID- 12654124 TI - Academic recognition of critical appraisal and systematic reviews in British postgraduate medical education. PMID- 12654125 TI - Practical skills acquisition during the houseman year. PMID- 12654126 TI - Self-directed learning activities of paediatric residents. PMID- 12654127 TI - What do we know about the anxieties of new clinical students? PMID- 12654129 TI - The effects of supraphysiological doses of corticosteroids in hypotensive liver failure. AB - BACKGROUND: In septic shock, supraphysiological doses of corticosteroids reduce norepinephrine requirements. We reviewed our experience of this treatment in hypotensive liver failure. METHODS: We retrospectively analysed 20 patients with liver failure who were treated with supraphysiological doses of hydrocortisone because of norepinephrine dependence. We compared their norepinephrine requirements, outcome, microbiology and incidence of gastrointestinal bleeding to an historical control group treated with norepinephrine but not corticosteroids. RESULTS: After 48 h of steroid treatment, the median norepinephrine dose was reduced (0.14 microg/kg/min to 0.08 microg/kg/min; P < 0.05) while the blood pressure over the same period of time did not change significantly (67.3 mm Hg to 70 mm Hg). Duration of ITU stay was longer in the steroid treated group (13.5 days vs 3 days; P < 0.05) but survival was similar in both groups. There were 23 episodes of positive bacterial cultures after norepinephrine was started in the steroid treated group, compared with 18 episodes in the control group. More of the positive cultures were due to resistant organisms in the steroid treated group (65% vs 17% in the control group; P < 0.002). There was no significant bleeding due to gastrointestinal inflammation in either group. CONCLUSIONS: Supraphysiological doses of corticosteroids reduce norepinephrine requirements in hypotensive liver failure. They do not improve survival but may extend time to find a suitable donor in those awaiting urgent liver transplantation. PMID- 12654130 TI - Prolonged cholestasis and ductopenia following gold salt therapy. AB - Hepatotoxicity, predominantly cholestatic, is a rare adverse effect of gold salt therapy, which usually completely resolves within a few months. We report the case of a female patient treated for rheumatoid arthritis, who had gold salt overdose, and in whom acute cholestatic hepatitis occurred three weeks after beginning of therapy. Evolution of gold concentration was followed in plasma and urine, as well as in cutaneous and liver dry tissue. Liver biopsy showed marked inflammatory changes of interlobular bile ducts that evolved towards ductopenia, which was responsible for prolonged cholestasis still present 15 months later. In addition, sialadenitis with sicca syndrome was noted six months after onset of the disease. The mechanism of hepatotoxicity was probably immunoallergic since liver lesions were associated with hypersensitivity syndrome including dermatitis and blood and tissue eosinophilia. This is the first report of gold salt hepatotoxicity with histological demonstration of cholangitis followed by ductopenia. PMID- 12654131 TI - Accumulating CD57 + CD3 + natural killer T cells are related to intrahepatic bile duct lesions in primary biliary cirrhosis. AB - BACKGROUND: Some populations of extrathymic T cells including natural killer T (NKT) cells are involved in autoimmune diseases. In particular, a deficiency of NKT cells has been implicated in human systemic sclerosis. Primary biliary cirrhosis (PBC) is an autoimmune liver disease characterized by the selective destruction of intrahepatic small bile ducts and the formation of granulomas. It is generally believed that cellular immune mechanisms, particularly T cells, cause the bile duct damage in PBC. METHODS: In this study, to investigate the unique population of extrathymic T cells in PBC, we examined the T cell receptor (TCR) V alpha 24-J alpha Q gene and CD57 + CD3 + cells corresponding to the major genotype and one of the phenotypes of NKT cells, respectively. RESULTS: By reverse transcription-polymerase chain reaction (RT-PCR), amplicons of the V alpha 24-J alpha Q gene fragment in liver tissues of PBC were found to be similar to those of control diseased livers, demonstrating that there was no distinct deficiency of V alpha 24-J alpha Q + NKT cells in PBC. Immunohistochemistry for CD57 and CD3 revealed the CD57 + CD3 + cells to be distributed in portal tracts and hepatic parenchyma in all cases studied, but the numbers of these cells were increased within portal tracts in PBC, in particular around injured interlobular bile ducts, compared to other diseased and normal livers. CONCLUSION: This indicates that an immune disturbance induced by a selective reduction of V alpha 24-J alpha Q + NKT cells is unlikely to occur in PBC. Inversely, it can be postulated that auto aggressive CD57 + CD3 + NKT cells are recruited to regulate the altered immunity of the periductal microenvironment in PBC. PMID- 12654132 TI - Reduced hepatic glycogen stores in patients with liver cirrhosis. AB - BACKGROUND: Patients with alcoholic liver cirrhosis have reduced hepatic glycogen stores but the mechanisms leading to this finding are not clear. METHODS: We therefore determined the hepatic glycogen content in patients with alcoholic (n = 9) or biliary cirrhosis (n = 8), and in control patients undergoing liver surgery (n = 14). All patients were in the postabsorptive state. In addition, we performed a morphometric analysis of the livers, and measured activities and mRNA expression of several enzymes involved in glycogen metabolism. Cirrhotic and control patients were similar regarding age and body weight. RESULTS: Cirrhotic patients had a reduced glycogen content per gram liver wet weight (17 +/- 11 versus 45 +/- 17 mg/g, P < 0.05), per milliliter hepatocytes (28 +/- 16 versus 52 +/- 21 mg/ml, P < 0.05) and per liver (28 +/- 17 versus 64 +/- 22 g, P < 0.05), the reduction being observed in both patients with alcoholic or biliary cirrhosis. Liver histology confirmed these findings and revealed that the decrease in liver glycogen in cirrhotic patients was not homogeneous across cirrhotic lobules. Activities of glycogen synthase and phosphorylase (total activity and active form) were not different between cirrhotic and control patients, whereas hepatic mRNA expression was decreased in cirrhotics by approximately 50%. The activity of glucokinase was decreased in cirrhotic as compared in control patients (0.06 +/- 0.30 versus 0.42 +/- 0.21 U/ml hepatocytes, P < 0.05), the reduction being observed in both patients with alcoholic or biliary cirrhosis. CONCLUSIONS: We conclude that patients with alcoholic or biliary cirrhosis have decreased hepatic glycogen stores per volume of hepatocytes and per liver. Decreased activity of glucokinase may represent an important mechanism leading to this finding. PMID- 12654133 TI - Hyaluronic acid and endothelial damage due to paracetamol-induced hepatotoxicity. AB - BACKGROUND: Damage to endothelial cells may be an important factor in the complications of acute liver failure, resulting in multi-organ failure. The aim of this study was to assess endothelial cell function in patients with severe hepatotoxicity due to paracetamol ingestion. PATIENTS AND METHODS: Fifty-eight patients with paracetamol-induced hepatotoxicity were studied for up to 7 days. Serum hyaluronic acid (HA), as a marker of hepatic sinusoidal endothelial cell function, was determined using an enzyme-linked binding assay. Plasma von Willebrand Factor, thrombomodulin and interleukin-8 were also determined using ELISA. RESULTS: Serum HA on admission was significantly increased (median 6777 ng/ml, range 24-50 967 ng/ml) as compared to normal controls (n = 10, median 21 ng/ml, range 0-50 ng/ml; P < 0.001). In non-survivors (n = 21) HA levels peaked on day 2 after admission (P = 0.044), and then decreased. In the survivors (n = 37) the levels of HA did not increase further. Plasma von Willebrand Factor, plasma thrombomodulin and serum interleukin-8 were significantly increased in the patients as compared to the normal controls (P < 0.001). Serum interleukin-8 was significantly higher in non-survivors in the first 2 days. CONCLUSIONS: Endothelial function is abnormal in paracetamol-induced hepatotoxicity. Damage to hepatic sinusoidal endothelial cells assessed by serum HA was greater in non survivors than survivors. PMID- 12654134 TI - Histological features associated with relapse after corticosteroid withdrawal in type 1 autoimmune hepatitis. AB - Relapse of type 1 autoimmune hepatitis after drug withdrawal may relate to incomplete histological improvement during corticosteroid therapy and/or persistence of pathogenic mechanisms. AIM: Determine the histological features prior to drug withdrawal that are associated with relapse in patients satisfying pre-established clinical, laboratory, and histological criteria for remission and relapsing after corticosteroid withdrawal. METHODS: One hundred liver tissue samples obtained immediately prior to corticosteroid withdrawal from 88 patients who had previously satisfied criteria for histological remission were reviewed retrospectively. RESULTS: Histological findings in the patients who relapsed were similar to those in the patients who sustained remission in regard to histological activity index (1.7 +/- 0.1 versus 1.6 +/- 0.2, P = 0.6), fibrosis score (2.6 +/- 0.3 versus 2.3 +/- 0.4, P = 0.5), and frequencies of interface hepatitis (36% versus 20%, P = 0.2), cirrhosis (21% versus 17%, P = 0.8), and normal or near normal tissue (9% versus 7%, P > 0.9). Only the presence of portal plasma cells was associated with relapse (31% versus 7%, P = 0.01). The positive predictability of portal plasma cell infiltration for relapse was 92%, but its sensitivity was only 31%. CONCLUSIONS: Portal plasma cell infiltration is predictive of relapse after drug withdrawal in tissue specimens already satisfying criteria for remission. Portal plasma cell infiltration may be indicative of an active antibody-dependent pathogenic mechanism. Its low sensitivity for relapse indicates the need for other complementary predictors of outcome. PMID- 12654135 TI - The molecular physiology of liver regeneration following partial hepatectomy. AB - The ability of the liver to regenerate after resection has been known for many years. Two reports from Germany in the late 1800s probably mark the introduction of the phenomenon into the scientific literature, but in the early 1900s the first reviews of this subject had appeared in the English literature. Predating these early scientific reports the legends from the Greek mythology described the fate of Prometheus. As punishment for defying Zeus and revealing the secret of fire to man, Prometheus was chained to a rock and each day had part of his liver ripped out by an eagle which, returning the following day, repeated the torture because his liver regenerated itself overnight. Although the speed of regeneration in the Greek legend is somewhat greater than that observed either clinically or in the laboratory, the myth does serve to emphasise the remarkable ability of the liver to repeatedly regenerate following repeated resections. This review aims to summarise the more recent literature concerning the early molecular events accompanying liver regeneration and to integrate this with the existing knowledge of this subject. PMID- 12654136 TI - Systematic review of the role of pre-oxygenation for tracheal suctioning in ventilated newborn infants. AB - Pre-oxygenation for endotracheal suctioning for mechanically ventilated infants is routine practice in many neonatal intensive care units. In the present systematic review the evidence to support its use is discussed and the authors conclude that no confident recommendations can be made from the results of this review. PMID- 12654137 TI - Children and war. AB - Children bear disproportionate consequences of armed conflict. The 21st century continues to see patterns of children enmeshed in international violence between opposing combatant forces, as victims of terrorist warfare, and, perhaps most tragically of all, as victims of civil wars. Innocent children so often are the victims of high-energy wounding from military ordinance. They sustain high-energy tissue damage and massive burns - injuries that are not commonly seen in civilian populations. Children have also been deliberately targeted victims in genocidal civil wars in Africa in the past decade, and hundreds of thousands have been killed and maimed in the context of close-quarter, hand-to-hand assaults of great ferocity. Paediatricians serve as uniformed military surgeons and as civilian doctors in both international and civil wars, and have a significant strategic role to play as advocates for the rights and welfare of children in the context of the evolving 'Laws of War'. One chronic legacy of contemporary warfare is blast injury to children from landmines. Such blasts leave children without feet or lower limbs, with genital injuries, blindness and deafness. This pattern of injury has become one of the post-civil war syndromes encountered by all intensivists and surgeons serving in four of the world's continents. The continued advocacy for the international ban on the manufacture, commerce and military use of antipersonnel landmines is a part of all paediatricians' obligation to promote the ethos of the Laws of War. Post-traumatic stress disorder remains an undertreated legacy of children who have been trapped in the shot and shell of battle as well as those displaced as refugees. An urgent, unfocused and unmet challenge has been the increase in, and plight of, child soldiers themselves. A new class of combatant comprises these children, who also become enmeshed in the triad of anarchic civil war, light-weight weaponry and drug or alcohol addiction. The International Criminal Court has outlawed as a War Crime, the conscription of children under 15 years of age. Nevertheless, there remain more than 300000 child soldiers active and enmeshed in psychopathic violence as part of both civil and international warfare. The typical profile of a child soldier is of a boy between the ages of 8 and 18 years, bonded into a group of armed peers, almost always an orphan, drug or alcohol addicted, amoral, merciless, illiterate and dangerous. Paediatricians have much to do to protect such war-enmeshed children, irrespective of the accident of their place of birth. Only by such vigorous and maintained advocacy can the world's children be better protected from the scourge of future wars. PMID- 12654138 TI - Predicting iron status in low birthweight infants. AB - OBJECTIVE: To determine the iron status of a selected group of low birthweight infants at approximately 9 months of age, and examine the feasibility of predicting iron status by examining the history of supplementary iron intake. METHODS: All live low birthweight infants recorded in the Dunedin Hospital Queen Mary Maternity Unit birth register who reached 9 months of age between November 1995 and September 1996 were eligible to participate. Infants were categorized into 'high' or 'low' iron intake groups depending on their consumption of infant formula or medicinal iron for one month prior to the study, and their iron status compared. RESULTS: Eighty-one infants of 73 mothers, with an average age of 10 months (range 8-13 months), participated. Thirty-three per cent (n = 27) were iron deficient: 19% (n = 15) had latent iron deficiency and 15% (n = 12) had iron deficiency anaemia. Those with a 'low' iron intake were 13-fold more likely to be iron deficient than infants with a 'high' iron intake (95% confidence interval: 4.4-41.5). Screening for iron deficiency using categories based on supplementary iron intake had a positive predictive value of 66% and a negative predictive value of 88%. CONCLUSIONS: The risk of iron deficiency was considerably greater for infants who had not received supplementary iron daily over the course of the previous month. Current preventative methods for avoiding poor iron status in this group of high risk infants are not effective. Screening for iron deficiency in low birthweight infants on the basis of iron intake from infant formula or medicinal iron provides a useful method for identifying infants whose iron status should be investigated. PMID- 12654139 TI - Occurrence of subclinical post-streptococcal glomerulonephritis in family contacts. AB - OBJECTIVE: There are few data in the published literature on the occurrence of subclinical post-streptococcal glomerulonephritis. In order to estimate the incidence of subclinical disease, 75 families of index cases with sporadic clinical post-streptococcal glomerulonephritis were screened for the presence of subclinical disease. METHODS: Three hundred and seventeen family contacts were investigated 1-7 days after the admission of the index cases. The diagnosis of subclinical disease was based on the presence of abnormal urinalysis, transitory hypocomplementaemia and increased antistreptolysin O titre. RESULTS: No cases of clinical/subclinical disease were detected among 147 parents. Abnormal urinalyses were found in 22.3% of sibling contacts. The incidence of nephritis among 170 siblings was 9.4% and the calculated ratio subclinical/clinical disease was 0.11. There were 16 siblings (9.4%) whose abnormal urinalyses could not be explained by appropriate tests; 11 of them had dysmorphic microhaematuria and significantly elevated antistreptolysin O titre. CONCLUSIONS: Sibling contacts have increased risk for the development of clinical/subclinical post-streptococcal glomerulonephritis compared with their parents. Sibling contacts with unexplained urinary abnormalities might have subclinical nephritis in evolution; their complement levels normalized before occurrence of nephritis in index cases. PMID- 12654140 TI - Slowing of growth in height and weight on stimulants: a characteristic pattern. AB - OBJECTIVE: The aims of the present study were to describe the growth pattern of children starting stimulant medication and to analyse the changes over time in height, weight and height velocity in a cohort of treated patients. METHODS: Retrospective review of growth data from files of all newly treated patients with attention-deficit/hyperactivity disorder in one paediatric practice. Forty-four boys and seven girls were treated for 6-42 months with either dexam-phetamine (n = 32) or methylphenidate (n = 19). RESULTS: During the first 6 months on stimulant medication 44 children (86%) had a height velocity below the age corrected mean and there was weight loss in 39 (76%). The height and weight standard deviation score (SDS) showed a progressive decline that was statistically significant after 6 and 18 months (P < 0.001, paired t-test). The height velocity was significantly attenuated for the first 30 months (P < 0.01), being lowest during the first 6 months. The mean height deficit during the first 2 years was approximately 1 cm/year. The change in weight SDS was 2.4 times the change in height SDS after 30 months on treatment with a significant correlation (Pearson's correlation coefficient r = 0.88, P < 0.001). CONCLUSIONS: Stimulant medication is associated with a decrease in height and weight SDS during the first 6-30 months with a characteristic pattern on the growth chart. PMID- 12654141 TI - Ultrasound grading of cerebral ventricular dilatation in preterm neonates. AB - OBJECTIVE: To compare three grading systems previously described for the measurement of cerebral ventricular dilatation in preterm neonates using ultrasound scans and then to compare these with expert clinical judgement. METHODS: Eighty-one preterm neonates <1500 g birthweight or < 32 weeks' gestation, with normal, mild, moderate or severely dilated ventricles recorded predominantly during the first 2 weeks of life were selected to ensure a broad representation of ventricular size. Their cranial ultrasound scans obtained nearest to 6 weeks of age were studied. The ventricular index, the diagonal width in the coronal plane and the ventricular height in the parasagittal plane were measured adjacent to the foramen of Monro and were used to grade the degree of dilatation. In 20 of these neonates, this grading was compared with grading based on expert clinical judgement. RESULTS: Grading according to the ventricular index defined 79% of the scans as normal. However, only 44% and 39% were considered to be normal according to vertical height and diagonal width, respectively. There was a high level of correlation between expert clinical judgement and grading based on ventricular height measurement (r = 0.812; area under receiver operating characteristic (ROC) curve = 0.922) and the diagonal width (r = 0.806; area under ROC curve = 0.935). Ventricular index correlated poorly with clinical judgement (r = 0.298; area under ROC curve = 0.604). CONCLUSIONS: The measurement of ventricular height and the measurement of diagonal width correlated well with expert clinical judgement. The ventricular index correlated poorly and was less sensitive to mild dilatation. There was no correlation between the ventricular index and the other two measures. The ventricular height and the diagonal width are more appropriate for assessing ventricular dilatation in preterm neonates. PMID- 12654142 TI - Birthweight distribution in southern Chinese infants with symptomatic congenital heart disease. AB - OBJECTIVE: Western infants with congenital heart disease have frequently been reported to have a low birthweight for gestational age. Studies in Asian infants seem to be lacking in this area. This is the first extensive study presenting the birthweight distribution of Chinese newborns with symptomatic congenital heart disease. METHODS: The birth data of 454, mainly southern-Chinese infants with symptomatic cardiovascular defects, born between 1990 and 1995 and admitted to Grantham Hospital, Hong Kong in 1994 and 1995, were analysed retrospectively. Infants with (non)-genetic syndromes or other major extracardiac malformations were excluded. RESULTS: Fifteen per cent of all newborns had a birthweight below the reference mean of - 2 Z-score. After correction for length of gestation, no significant difference could be detected in birthweight between the cyanotic and acyanotic groups, nor between the different haemodynamic disturbances. Infants with atrial or ventricular septal defects, tetralogy of Fallot, pulmonary atresia with ventricular septal defect, heart with univentricular atrio-ventricular connection or double outlet right ventricle showed significant birthweight deficits. Transposition of the great arteries was not related to being small for gestational age. CONCLUSIONS: Similar to Western infants, prenatal growth impairment was a common feature in Chinese infants with symptomatic congenital heart disease. The birthweight distribution in Chinese might be comparable to that in Western populations. Exceptions are possibly the high frequency of low birthweight in Chinese newborns with atrial septal defect or a single-ventricle abnormality. Further studies on fetal anthropometry and haemodynamics are necessary to provide insight into the relation between cardiovascular malformations and being small for gestational age. PMID- 12654143 TI - Effect of postural supports on neuromotor function in very preterm infants to term equivalent age. AB - OBJECTIVE: To determine the effect of a postural support nappy and/or a postural support roll on neuromotor function in very preterm infants when nursed prone to term equivalent age. METHODS: A randomized observer blind controlled trial of 123 very preterm infants was conducted in the neonatal intensive care unit of the sole tertiary referral centre in Western Australia. Infants were stratified by gestational age (< 29 weeks or 29-30 weeks), then randomized into one of three intervention groups: postural support nappy, postural support nappy and postural support roll, or disposable nappy and postural support roll. Interventions started when infants were stable and ceased when routine side-lying commenced. Measurements of shoulder and hip posture were performed pre-intervention, 5 weeks post-intervention and term postmenstrual age. RESULTS: Infants nursed with a postural support roll and a postural support nappy demonstrated improved hip posture to term equivalent age compared with infants nursed with either a postural support roll only, or a postural support nappy only. Infants nursed with a postural support roll either with or without a postural support nappy demonstrated improved shoulder posture to term equivalent age. CONCLUSIONS: Combined use of a postural support roll and a postural support nappy while very preterm infants are nursed prone improves hip posture up to term postmenstrual age. Use of a postural support roll improves shoulder posture up to term equivalent age. PMID- 12654144 TI - Predicting the need for ventilatory support in neonates 30-36 weeks' gestational age. AB - OBJECTIVES: To determine antenatal factors associated with the need for ventilatory support in babies born between 30 and 36 weeks gestational age and use this information to help referring obstetricians decide which mothers need antenatal transfer. METHODS: Babies born at Royal Prince Alfred Hospital at 30-36 weeks' gestation inclusive between January 1992 to December 1999 were identified in the obstetric and neonatal databases. Information for a wide range of antenatal factors and respiratory outcomes was extracted. Babies with major congenital anomalies were excluded. Statistical analysis using spss for Windows was then undertaken. RESULTS: Data were available for 3102 babies. On univariate analysis, seven factors were significant. Lower gestational age, absence of labour, Caesarean section, antepartum haemorrhage, breech presentation and hypertensive disease of pregnancy were associated with increased risk, while threatened premature labour was protective. On logistic regression analysis, gestational age and absence of labour dominated the model. Other significant factors in the model had a relatively minor impact. CONCLUSIONS: The risk of needing ventilatory support according to antenatal risk factors is described for infants born between 30 and 36 weeks. Gestational age and absence of labour were found to be the major determinants of risk. Delivery without labour increased the risk by the equivalent of about 2 weeks of gestation. For example, to avoid a risk > 20%, the cut off is 33 weeks for mothers who labour, but is increased to 35 weeks when there is no labour. Our results should be interpreted with caution, as the patients in a tertiary obstetric unit may not be typical of the wider community. PMID- 12654145 TI - Peak inspiratory flows in children with cystic fibrosis. AB - OBJECTIVE: To show that in children with moderately severe cystic fibrosis lung disease: (i). inspiratory flow may be reduced; and (ii). peak inspiratory flow may be predicted from height, expiratory flow analysis or body mass index. METHODS: All children attending the Royal Children's Hospital, Melbourne, between May and July, 2001 who had cystic fibrosis, were aged > 5 years, were able to perform spirometry reproducibly and who had a forced expiratory volume in 1 s < 60% predicted were prospectively enrolled. Height, weight, peak inspiratory flow, forced expiratory volume in 1 s and forced vital capacity were recorded. Linear regression analysis was performed. RESULTS: The age range was 9.4-19.9 years. Sixteen boys and 11 girls were studied. All children had a peak inspiratory flow > 0.5 L/s. There was a significant relationship between peak inspiratory flow and forced vital capacity (R2 = 0.50) especially in boys (R2 = 0.65). In boys, peak inspiratory flow was significantly related to forced expiratory volume in 1 s (R2 = 0.47). There was no relationship between peak inspiratory flow and predicted values of expiratory flow, age, height, weight or body mass index. Logistic regression was used to predict the probability that peak inspiratory flow was < 2.0 L/s for a given forced vital capacity. If the forced vital capacity is > 2.5 L, peak inspiratory flow is likely to be > 2.0 L/s. CONCLUSIONS: In children with significant cystic fibrosis lung disease, peak inspiratory flow is likely to be > 0.5 L/s, which is required to activate dry powder inhalers. If the forced vital capacity is < 2.5 L, the peak inspiratory flow may be < 2.0 L/s, and a metered dose inhaler and spacer should be considered. Further studies that investigate the relationship between expiratory flow and peak inspiratory flow against an internal resistance are needed. PMID- 12654146 TI - Is pentoxifylline therapy effective for the treatment of acute rheumatic carditis? A pilot study. AB - OBJECTIVE: The aim of the present study was to determine whether pentoxifylline has a beneficial effect on the treatment of rheumatic carditis. METHODS: A total of 33 children between the ages 6 and 16 were studied in two groups. The first group (5 boys, 10 girls, mean age: 12.2 +/- 2.9 years) was treated with steroid plus pentoxifylline and the second group (6 boys, 12 girls, mean age; 11.6 +/- 2.8 years) was treated with steroid only for 3-6 weeks until the acute-phase reactants became normal. At admission and on the 7th, 30th, and 90th days of the treatment, laboratory studies including white blood cell count, erythrocyte sedimentation rate, C-reactive protein, throat culture and cytokines (interleukin 1alpha, tumour necrosis factor-alpha) were performed. Cardiac evaluation with chest X-ray, electrocardiography and echocardiography was performed in all patients. In the control group (12 boys, 3 girls, mean age; 10.7 +/- 3.2 years) all parameters were evaluated once only. RESULTS: In both groups, the similar white blood cell count was significantly decreased on the 90th day, and there was no significant difference between the two groups. C-reactive protein, erythrocyte sedimentation rate and interleukin-1alpha were significantly decreased on the 30th and 90th days. In the first group (treated with steroid plus pentoxifylline), the cardiothoracic index was significantly greater at the beginning of the therapy. In the first group, tumour necrosis factor-alpha became normal on the 30th day and in the second group, tumour necrosis factor-alpha became normal on the 7th day of therapy. For all parameters, there was no significant difference between the two groups with respect to the type of therapy used. CONCLUSION: The present study showed that pentoxifylline plus steroid treatment has no beneficial effects on the treatment of acute rheumatic carditis when compared with steroid alone. PMID- 12654147 TI - Ethical issues in paediatric rehabilitation. PMID- 12654148 TI - Brain death. PMID- 12654149 TI - Superior sagittal sinus thrombosis in a child with Down syndrome. PMID- 12654150 TI - Development of psoriatic lesions during acute and convalescent phases of Kawasaki disease. AB - A 7-month-old infant developed a discrete pustular rash confined to both soles during the acute phase of Kawasaki disease. Histological examination of a pustular lesion from the sole of a foot showed subcorneal neutrophilic microabscesses, psoriasiform acanthosis with a thin granular layer and mononuclear perivascular infiltrates in the upper dermis, consistent with psoriasis. Following the standard treatment with intravenous gamma globulin, the initial symptoms and signs of Kawasaki disease resolved completely. Eight weeks later, psoriasiform plaques appeared on both cheeks and on the extensor surfaces of the forearms and legs. Skin biopsy from one of these lesions revealed psoriasiform epidermal hyperplasia, focal parakeratosis and dilated papillary capillaries. The patient was treated with mild-potency topical steroids that resulted in rapid and complete resolution of the skin lesions. Concurrent association of psoriatic skin lesions and Kawasaki disease might not be incidental and could stem from a common pathogenetic mechanism induced by superantigens. PMID- 12654151 TI - Evolving hypopituitarism following cranial irradiation. AB - Increasing survival after treatment for childhood cancer in recent years has left many patients with long-term sequelae. Following cranial irradiation, changes in hypothalamic-pituitary function evolve over several years and multiple hormone deficiencies are frequently found. In the present study we describe a boy whose initial presentation with a cerebral tumour included central precocious puberty. He was followed for more than 15 years and sequentially developed deficiencies of growth hormone, thyroid-stimulating hormone, gonadotrophins and adrenocorticotropic hormone after high-dose cranial irradiation. Long-term endocrine follow up of such children is essential for the early diagnosis and optimal management of hormone deficiencies. PMID- 12654152 TI - Concurrent HIV infection and neuroblastoma. AB - An infant is reported who presented with Pneumocystis carinii pneumonia, secondary to HIV infection, diagnosed at 3 months of age, and who had concurrent paravertebral neuroblastoma. Although neuroblastoma cell lines support the growth of HIV in vitro, this is the first report of a clinical association between HIV infection and neuroblastoma. Although we do not think the conditions are causally linked, we report the case to raise awareness of a possible association between HIV and neuroblastoma. The case also raises the importance of starting antiretroviral treatment with three drugs. PMID- 12654153 TI - Colonic atresia and associated anomalies. PMID- 12654161 TI - Hypoxia-induced megaloblastosis in vitamin B12-deficient rats. AB - In rats, in contrast with human subjects who develop megaloblastic anaemia due to vitamin B12 deficiency, haematological abnormalities with anaemia were not observed under normoxic conditions even though plasma vitamin B12 concentration was reduced to <15 % of a normal concentration by depleting dietary vitamin B12. To elucidate whether erythropoiesis was affected by vitamin B12 deficiency in rats, these vitamin B12-deficient rats were exposed to hypoxia (10.5 % O2) to stimulate erythropoiesis. In the vitamin B12-sufficient control rats, erythrocyte count was significantly (P<0.05) increased 1 week after starting the hypoxic exposure. However, the hypoxia-induced erythropoiesis was affected by vitamin B12 deficiency, and no significant increase in the erythrocyte count was observed even after 6-week exposure to hypoxia in the vitamin B12-deficient rats. In the vitamin B12-deficient rats in hypoxia, erythrocytes became abnormally enlarged, and haemoglobin concentration in peripheral blood was increased in proportion to the increase of mean corpuscular volume. However, the level of the increase in the haemoglobin concentration was significantly (P<0.05) lower in the vitamin B12 deficient rats compared with that in the -sufficient controls. In addition, in the vitamin B12-deficient rats, in contrast to the -sufficient rats, serum erythropoietin concentration was not normalized even after 6-week exposure to hypoxia. These results indicate that a megaloblastic anaemia-like symptom is induced when the vitamin B12-deficient rats are exposed to hypoxia. PMID- 12654160 TI - Megaloblastic anaemia in vitamin B12 deficiency. PMID- 12654162 TI - Supplementation of barley straw with Sesbania pachycarpa leaves in vitro: effects on fermentation variables and rumen microbial population structure quantified by ribosomal RNA-targeted probes. AB - Tropical livestock is often maintained on roughage-based diets deficient in N, and therefore requires supplementation with protein-rich substrates to achieve reasonable production levels. The optimum inclusion rate of a potential supplement is usually determined by in vivo feeding trials or by in vitro incubation of the diet components to estimate the feed value of the complete diet. The present work simulates a supplementation experiment in vitro, by incubating a pure roughage (barley straw), a pure supplement (Sesbania pachycarpa leaves) and mixtures of the two, with increasing inclusion levels of the supplement, in a short-term batch incubation system. Fermentation kinetics were followed by the release of fermentation endproducts (gas and short-chain fatty acids). Microbial biomass was estimated using ribosomal (r) RNA as internal marker for bacteria and eukaryotes separately. Cell-wall-degrading subpopulations were quantified by hybridisation with taxon-specific oligonucleotide probes targeting Chytridiomycetes, Fibrobacter spp., Ruminococcus albus and R. flavefaciens. Carboxymethylcellulase (CMCase) was assayed as an indicator for cell-wall-degrading activity. The addition of S. pachycarpa leaves stimulated fermentation in all cases. Gas production, and especially rRNA concentration, showed clear maxima at 40 % S. pachycarpa inclusion, rates that significantly exceeded the values interpolated from the incubations of the pure substrates. Short-chain fatty acid yield changed only slightly, but in the same way. The analysis of the microbial population structure showed that the positive effects were mainly mediated through enhanced growth of Ruminococcus spp. Increasing proportions of S. pachycarpa leaves in the diet led to a drastic decline in the total eukaryotic population. This points to a defaunation, which may also have added to the positive effects. The eukaryotic subpopulation of the rumen fungi were affected to a lesser degree. Although the cell-wall-degrading organisms showed positive responses to the supplementation, the CMCase activity was not affected significantly by the supplementation. The present work shows that it is possible to predict optimum inclusion levels for a new feed supplement in vitro and thus reduce in vivo experiments. It was also demonstrated that true supplementation effects occur particularly for the microbial biomass production, which is the primary source of amino acids for the ruminant animal. The analysis of microbial population structure in context with conventional metabolic measurements adds valuable information to interpret the observed effects on production-related variables. PMID- 12654163 TI - Influence of sources of dietary vitamin E on the maternal transfer of alpha tocopherol to fetal and neonatal guinea pigs as determined by a stable isotopic technique. AB - The accepted biological potencies of vitamin E (United States Phamacopeia, 1985) for 1 mg all-rac-alpha-tocopheryl acetate (synthetic form) is 1.00 IU and that of 1 mg (RRR)-alpha-tocopheryl acetate (natural form) is 1.36 IU. In the present study, a stable isotopic (2H) technique was employed to evaluate the bioavailability of natural v. synthetic forms of vitamin E and to determine whether the potency of the forms is the stated relationship of 1.36:1.00 (RRR) alpha-tocopheryl acetate:all-rac-alpha-tocopheryl acetate. Sixty female in-bred guinea pigs received either 40 or 80 mg vitamin E/kg diet with equal levels of (RRR)-alpha-tocopheryl acetate and all-rac-alpha-tocopheryl acetate throughout gestation and lactation. At late-term pregnancy (day 50 or 60) and during early lactation, dams and their corresponding fetuses or neonates were killed and various tissues collected for subsequent alpha-tocopherol analysis. Vitamin E analysis of fetal and neonatal tissues indicated a substantial transfer of 2H labelled alpha-tocopherol across the placenta and through the mammary gland. Total alpha-tocopherol concentrations were significantly influenced by tissue type and dose level, but not by stage of gestation or lactation. The relative bioavailability (d3:d6) across fetal and neonatal tissues was on average 1.81:1.00, with a range from 1.62:1.00 to 2.01:1.00. Maternal tissues had a mean ratio of 1.77:1.00. A higher relative bioavailability (P2 g/d) known to increase lipid peroxidation. In contrast, very low doses, that may prevent lipid peroxidation, might affect the immune system differently. To study the latter hypothesis further, we investigated whether the supplementation of healthy elderly people with very low doses of marine oil (MO), a docosahexaenoate (DHA)- and eicosapentaenoate (EPA)-rich triacylglycerol, was able to affect lymphocyte proliferation and biochemical markers known to be altered with age. In a randomized, double-blind design, twenty healthy elderly subjects were assigned to a placebo group (600 mg sunflower oil/d) or to a group consuming 600 mg MO/d providing 150 mg DHA + 30 mg (EPA) for 6 weeks. At day 42, the proliferative responses of lymphocytes to several mitogens were significantly (P<0.01) decreased in the MO group compared with control values. This was accompanied by a slight lowering of their cytosolic cyclic nucleotide phosphodiesterase (PDE) activity, a marked and significant (P<0.05) increase of their particulate PDE activity (+56-57 %) and a slight but significant (P<0.05) increase in cyclic nucleotide intracellular levels. At the same time, the glutathione peroxidase activity was markedly and significantly (P<0.01) depressed in the MO group. None of these modifications could be seen in the placebo group. Collectively, these results demonstrate that even very low doses of n-3 fatty acids are sufficient to affect the immune responses of elderly subjects. PMID- 12654173 TI - Effect of a methionine-supplemented diet on the blood pressure of Wistar-Kyoto and spontaneously hypertensive rats. AB - The objectives of the present work were to evaluate the effect of a methionine supplemented diet as a model of hyperhomocysteinaemia on the systolic blood pressure (BP) and vasomotor functions of aortic rings in Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR). WKY and SHR rats, randomised into four groups, were fed a normal semisynthetic diet or a methionine (8 g/kg) supplemented diet for 10 weeks. Systolic BP was measured non-invasively. At the end of the experiment, plasma homocysteine, methionine, cysteine and glutathione levels were determined. Vasoconstriction and vasodilatation of aortic rings were measured. The methionine-supplemented diet induced a significant increase in plasma homocysteine and methionine concentration in both WKY and SHR rats, an increase in plasma cysteine concentrations in WKY rats and an increase in the glutathione concentration in SHR. The systolic BP of WKY rats fed the methionine supplemented diet increased significantly (P<0.01), whereas systolic BP was reduced in SHR. An enhanced aortic responsiveness to noradrenaline and a decreased relaxation induced by acetylcholine and bradykinin were observed in the WKY rats fed the methionine-enriched diet. In SHR, the bradykinin-induced relaxation was reduced, but the sodium nitroprusside response was increased. In conclusion, a methionine-enriched diet induced a moderate hyperhomocysteinaemia and an elevated systolic BP in WKY rats that was consistent with the observed endothelial dysfunction. In SHR, discrepancies between the decreased systolic BP and the vascular alterations suggest more complex interactions of the methionine enriched diet on the systolic BP. Further investigations are needed to understand the paradoxical effect of a methionine-rich diet on systolic BP. PMID- 12654174 TI - [Bioinformatic analysis of glioma development relative genes]. AB - BACKGROUND & OBJECTIVE: Exploiting the transcriptional regulation mechanism by microarray and bioinformatics is an important method at genomic research field, and it is better than previous methods, by which we can easily analyze the gene expression regulative networks at genomic level. This study was designed to analyze the glioma gene expression profiles by clustering and bioinformatic retrieving to seek some tumor development relative genes that can be cloned for the purpose of function research in the future. METHODS: Firstly, we analyzed the glioma gene expression profiles of 16,363 genes by clustering, and chose 368 genes which expression differences were greater than 2 folds and the variances of 2 dots were smaller than 0.33. We selected 2 groups of genes that were expressed similarly. One group (11 cases) was upregulated with the glioma development and another (6 cases) was downregulated with the glioma development. Secondly, we determined the genomic information about those 17 genes and exploited their association with the development of gliomas. RESULTS: Two groups of genes were expressed similarly during the glioma development, one group was upregulated with the development of gliomas, and another was downregulated. Bioinformatic analysis showed that 3 of those genes (X55987, M85085, and AB011097) might be important tumor relative genes. CONCLUSION: By bioinformatics and microarray technologies, we found 3 genes, X55987(EDN, eosinophil-derived neurotoxin), AB011097 (ARTS-1, TNF receptor shedding regulator),and M85085(CStF, cleavage stimulation factor), which might be potential key tumor development relative genes that can be developed for therapy targets in the future. PMID- 12654175 TI - [Cloning, sequencing and bioinformatics analysis of a new tumor suppressor gene ndr2 from mouse]. AB - BACKGROUND & OBJECTIVE: Ndr2 (N-myc down stream regulator) gene in human is a new gene cloned with the human adult whole brain cDNA as template in 1999, which accession number is AF159092 in GenBank. Locating backward position of the N-myc gene in human chromosome, this gene was named Ndr2 gene. The previous experimental results showed Ndr2 gene probably is a tumor suppressor gene. To research the function of Ndr2 gene, the authors cloned the genomic sequence of ndr2 from mouse. METHODS: To clone Ndr2 genomic sequence by reverse transcription polymerase chain reaction(RT-PCR) with the mouse genome library as template; automatic sequencing was performed using 310 Genetic Analyzer; homogeneous analysis was made using GenBank BLAST; open reading fragment(ORF) analysis was made using PC Gene and ORF Finder; domain analysis was made using ProDom system. RESULTS: A fragment (about 3310bp,identified by agarose gel electrophoresis) was obtained using RT-PCR with the mouse genome library as template. The fragment was cloned in pMD18-T vector. BLAST analysis showed that the sequence was highly homogeneous (with the homogeneity rate of 91.4%) with Ndr2 gene in human and non homogeneous with genomic sequence database in mouse. ORF analysis showed that there was a complete coding region in it, which including 8 extrons and 7 introns; it can interpret a protein containing about 200 amino acid residuals. ProDom analysis showed there was a domain like acyl carrier protein(ACP) in it. CONCLUSION: The authors cloned Ndr2 gene in mouse and proved that the sequence is a new genome sequence in mouse genomic sequence database. At present, the genome sequence has been submitted to GenBank(the accession number: AY151387). PMID- 12654177 TI - [Analysis of gene expression patterns of leukemia K562 cells after cytochalasin B treatment]. AB - BACKGROUND & OBJECTIVE: The advanced technique of DNA microarray makes it possible to monitor the expression of thousands of genes simultaneously in one hybridization experiment. This technique accelerates demonstration of anti-tumor drug mechanisms and discovery of new drug targets. This study was designed to investigate the differential gene expression of K562 cells after cytochalasin B treatment using cDNA microarray. METHODS: Restriction display polymerase chain reaction (RD-PCR) products of 277 human genes were spotted on a glass slide in microarray. K562 cells grew in RPMI 1640 medium with 10 microg/ml cytochalasin B. After 24 hours, the total RNA was isolated from K562 cells, and mRNA was purified. Both mRNA from the treated K562 cells and the controlled K562 cells were reversely transcribed into cDNA and labeled with two different fluorescence dyes: Cy5 or Cy3, using a method of restriction digestion and PCR labeling (RD PCR). The probes were hybridized to the cDNA microarrays. After high-stringent washing,the cDNA microarray was scanned for the fluorescent signals and showed difference between the two cells. RESULTS: Among the 277 target genes, 18 down regulated genes were identified after cytochalasin B treatment. CONCLUSION: There is a consistent tendency toward lower-expressed genes in partial K562 cells after cytochalasin B treatment. Most down-regulated genes were correlated with cell proliferation, signal transduction, and transcription factor. PMID- 12654176 TI - [Analysis of characteristics of gene expression in pericancerous stromal cells of nasopharyngeal carcinoma by cDNA array]. AB - BACKGROUND & OBJECTIVE: The mechanism of how stromal cell play an important role in nasopharyngeal carcinogenesis is now a hotspot. This study was designed to elucidate the possible mechanism of stromal cell in the occurrence and progression of nasopharyngeal carcinoma (NPC) through the analysis of the characteristics of gene expression in pericancerous stromal cells of NPC by cDNA array. METHODS: The atlas human select tumor arrays were used to compare the expression profiles between NPC tissue and NPC cell lines. RESULTS: Pericancerous stromal cells in NPC expressed at least 40 genes specifically. CONCLUSION: The specific expression of these genes in pericancerous stromal cells provides energy materials for growth of NPC cells; furthermore, it can accelerate the degradation of extracellular matrix, thus promoting the metastasis of NPC cells. PMID- 12654178 TI - [Relationship between Helicobacter pylori infection and proliferation and apoptosis of gastric epithelial dysplasia cell]. AB - BACKGROUND & OBJECTIVE: Gastric epithelial dysplasia is the precancerous lesion of gastric cancer. However, the mechanism that dysplasia evolves to malignancy is not clear. In order to clarify the relationship between Helicobacter pylori (HP) infection and its virulence factor and changes of cell kinetics of dysplasia, the authors measured the changes of proliferation and apoptosis and the status of HP infection. METHODS: A total of 117 gastric mucosal biopsy specimens were enrolled, including 12 of chronic superficial gastritis (CSG) and 105 of dysplasia. Dysplasia samples were divided into two groups: 35 of high-grade dysplasia [carcinogenesis group (n=30), regression group (n=5)], 70 of low-grade dysplasia [carcinogenesis group (n=18), regression group (n=52)]. The expression of proliferating cell nuclear antigen (PCNA) was measured by immunohistochemical staining; cell apoptosis was determined by terminal deoxynucleotidyl transferase mediated nick end labeling(TUNEL); the status of HP infection was detected by polymerase chain reaction(PCR) with specific primers of urea A and cagA gene. RESULTS: There was no significant difference of HP infection between dysplasia and CSG(84.76% vs. 83.33%), but CagA-positive strain infection rate in dysplasia was slightly higher than that in CSG (85.39% vs. 60.00%). Proliferation indexes(PI) in the patients with HP infection and CagA(+) strain infection were higher than that in the patients without HP infection and CagA(-)strain, respectively (P< 0.05). PI was positively associated with the status of HP and CagA(+) strains infection (P< 0.05). AI/PI ratio (AI: apoptosis index) was negatively associated with CagA-positive strain infection (P< 0.05). CONCLUSION: Gastric epithelial dysplasia cells have abnormal changes in PI and AI when it evolves to malignancy, and the abnormal cell kinetics is partly correlated with HP and CagA(+) strain infection. So treatment of HP infection may produce a good result for the evolution of dysplasia. PMID- 12654179 TI - [Expression of 67kD laminin receptor in human hepatocellular carcinoma cells]. AB - BACKGROUND & OBJECTIVE: Previous study of our laboratory showed that 67kD laminin receptor (67LR) from SMMC-7721 hepatocellular carcinoma (HCC) cells had higher binding affinity with laminin than that from L-02 normal hepatic cells; however, the expression level of 67LR in HCC cells and normal hepatic cells was still unknown. The aim of this study was to investigate the relationship between laminin binding activity and the expression of mRNA and protein of 67kD laminin receptor in SMMC-7721,HepG2 HCC cells, and L-02 hepatic cells. METHODS: (1) The binding of laminin with three kinds of cells was quantified by (131)I-labeled laminin. (2)The expression of 67kD laminin receptor was determined by the reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry. RESULTS: (1) Under the same condition, the amount of radiolabelled laminin bound to SMMC-7721, HepG2 HCC cells was 17.54+/-0.49 ng per 10(5) cells and 11.18+/-0.53 ng per 10(5) cells, respectively, which were significantly higher than that to L-02 hepatic cells (8.36+/-0.48 ng per 10(5) cells). The binding of laminin with HCC cells is specific and saturable, and the cells bound exogenous laminin with high affinity. (2) 67 LR showed notably increased expression on the surface of L-02 hepatic cells (55.3%), compared with SMMC-7721 HCC cells (34.7%) by flow cytometry. HepG2 did not express membrance 67 LR. (3) The expression of 67LR mRNA in SMMC-7721 and HepG2 HCC cells was higher than that in L-02 cells. CONCLUSION: The conflict of laminin binding activity and the expression of 67 LR in the three cell lines suggested SMMC-7721 HCC cells may express 67 LR with higher laminin affinity than L-02 hepatic cells, and other laminin receptors were involved in laminin binding. PMID- 12654180 TI - [Apoptosis of human gastric cancer cell induced by photochemical riboflavin]. AB - BACKGROUND & OBJECTIVE: Photochemical riboflavin, the production of reactive oxygen species (ROS), has been reported having cytotoxity on some cancer cells. The aim of this study was to investigate the effect of photochemical riboflavin on inducing apoptosis in human gastric cancer cell line MGC80-3. METHODS: Trypan blue exclusion method, Giemsa staining, DNA electrophoresis, DNA quantification, Western blot analysis, and flow cytometry were conducted to determine the effect of photochemical riboflavin on cell survival, morphology, DNA fragmentation, gene expression, and cell cycle arresting. RESULTS: The cell viability dropped down according to the riboflavin concentration and treating time. Exposure of the cells in 20 micromol/L riboflavin for 24 hours resulted in typical apoptotic morphology and G(2)/M arresting. As MGC80-3 cells were separately treated with 10, 20, 30, and 40 micromol/L photochemical riboflavin, DNA electrophoresis showed that the ladder bands, a typical feature of apoptotic cell, appeared in the groups treated by over 20 micromol/L photochemical riboflavin. The efficiency rates of DNA fragmentation were 35.4%, 54.1%, 70.6%, and 86.8%, respectively. Western blot analysis showed that the expression of apoptosis related proteins p53, C-myc, and Bax were up-regulated whereas the expression of Bcl-2 was down regulated. CONCLUSION: These results indicate that photochemical riboflavin has high efficiency of inducing cell apoptosis on MGC 80-3 cells in vitro and there is a correlation between apoptosis and G(2)/M arresting. PMID- 12654181 TI - [Effects of lovastatin on proliferation and gap junctional intercellular communication of human breast cancer cell MCF-7]. AB - BACKGROUND & OBJECTIVE: Lovastatin,an inhibitor of endogenous cholesterol biosynthesis,has been widely used in the clinical treatment of hypercholesterolemia.Recently,lovastatin has been paid more attention for its wide-range effects on human cancer cells; however,the detail mechanisms of its anti-cancer effects are not yet understood. This study was designed to investigate the effects of lovastatin on proliferation and gap junctional intercellular communication (GJIC) of MCF-7 human breast cancer cells. METHODS: After treated with lovastatin at dosages of 4,8,16 micromol/L for 1-3 days,the cell differentiation was examined with nitroblue tetrazolium (NBT) reduction test;the proliferation and distribution of cell cycles were examined with flow cytometry (FCM). Meanwhile,GJIC of MCF-7 cells was observed using the scrape loading and dye transfer(SLDT) technique. RESULTS: Lovastatin could inhibit the proliferation of MCF-7 cells significantly and 75.80 percent of cells were inhibited after treated with 16 micromol/L lovastatin for 72 hours (P< 0.05). Meanwhile, lovastatin could arrest MCF-7 cells in the G(0)/G(1) phase of cell cycle and 80 percent of cells were arrested in G(0)/G(1) phase after treated with lovastatin for 72 hours. Furthermore, lovastatin could induce the differentiation of MCF-7 cells (P< 0.01) and up-regulate GJIC in MCF-7 cells. After treated with 16 micromol/L lovastatin for 72 hours, transfer of LY fluorescence could reach 4 5 rows of cells from the scraped line. However, apoptosis in MCF-7 cells was not obvious. All these effects of lovastatin were in a dose-and time-dependent manner. CONCLUSION: It suggests that lovastatin has the capabilities of inhibiting proliferation, arresting MCF-7 cells at G(0)/G(1) phase of cell cycle and inducing differentiation. These effects of lovastatin maybe correlate with lovastatin promoting GJIC function in MCF-7 cells. PMID- 12654182 TI - [Significance of expression of epidermal growth factor (EGF) and its receptor (EGFR) in chronic cholecystitis and gallbladder carcinoma]. AB - BACKGROUND & OBJECTIVE: Epidermal growth factor(EGF) and its receptor(EGFR) play an important role in tumorigenesis. Cholelithiasis and cholecystitis were closely related to gallbladder carcinoma. This study was designed to investigate the relationship between the expression of EGF and EGFR in chronic cholecystitis and gallbladder carcinoma. METHOD: SABC immunohistochemistry was used to determine the expression of EGF, EGFR, and proliferating cell nuclear antigen (PCNA) in surgically resected specimens of 41 gallbladder carcinomas, 26 simple hyperplasia tissues, 14 dysplasia tissues,and 10 normal gallbladder tissues. RESULTS: The positive expression rates of EGF and EGFR were higher in gallbladder carcinomas(63.4%,70.7%) and dysplasia tissues (71.4%,85.7%) than in simple hyperplasia tissues (15.4%,27%) and normal gallbladder tissues (0%,0%)(P< 0.01); PCNA counting score tended to increase with the severity of mucosa change degree(P< 0.01), which were 1.0 in normal tissues, 1.0 in simple hyperplasia tissues, 1.9288+/-0.9972 in dysplasia tissues,and 3.0488+/-0.669 in gallbladder carcinomas. Significantly positive correlation were shown between EGF,EGFR, and PCNA(P< 0.05). There was no correlation between EGF, EFGR, and PCNA expression and tumor TNM stage(P>0.05). CONCLUSION: Expression of EGF and EGFR is involved in the gallbladder carcinogenesis, and is related to high activity of cell proliferation. PMID- 12654183 TI - [Significance of phosphorylation of mitogen-activated protein kinase and signal transducer and activator of transcription 3 and cyclin D1 protein expression in Hodgkin's lymphomas]. AB - BACKGROUND & OBJECTIVE: Signal cascades of mitogen-actived protein kinase(MAPK) and signal transducer and activator of transcription 3 (Stat3) are two main signal transduction pathways which were associated with cell proliferation and malignant transformation.MAPK and Stat3 proteins are activated by phosphorylation. The biological effects which are caused by multiple cytokines produced by Hodgkin's lymphoma(HL) cells are mediated through MAPK and Stat3 signal pathways. This study was designed to investigate significance of MAPK and Stat3 phosphorylation(p-MAPK and p-Stat3) and cyclin D1 protein expression in HL. METHODS: SP immunohistochemistry was used to detect expression of p-MAPK, p Stat3, and cyclin D1 protein in 45 cases of HL of various types. RESULTS: The expression positive rates of p-MAPK, p-Stat3, and cyclin D1 proteins were 73.3%(33/45),64.4%(29/45), and 68.9%(31/45), respectively. The positive expression levels of p-MAPK and cyclin D1 protein gradually increased(P< 0.05), whereas that of p-stat3 had no significant difference(P >0.05) in four subsets(LR:lymphocyte-rich classical type; NS:nodular sclerosis type; MC:mixed cellularity type; LD:lymphocyte depletion type) of all cases. The expression of p MAPK was positively related to that of cyclin D1 protein (r(s)=0.7254,P< 0.01), but the expression of p-Stat3 was not related to that of cyclin D1 protein (r(s)=0.2197,P >0.05). CONCLUSION: The data suggest that activation of MAPK may play an important role in genesis and progression of HL, but Stat3 activation is not associated with the progression of HL. MAPK may induce overexpression of cyclin D1 protein and results in persistent proliferation of RS/H cells in genesis and development of HL. PMID- 12654184 TI - [Inhibition of triclosan to fatty acid synthase from goose uropygial glands and human breast cancer cells in vitro]. AB - BACKGROUND & OBJECTIVE: It has been indicated that fatty acid synthase (FAS) is abnormally overexpressed in human breast cancer compared with normal human tissue. Inhibition of FAS induces apoptosis of human breast cancer cells. The aim of this study was to observe the inhibition of triclosan on FAS from goose uropygial glands for establishing the method and to study the inhibition of triclosan on FAS from human breast cancer SKBr3 cells in vitro. METHODS: The goose uropygial glands FAS was purified by ultra-centrifugation and Superdex PG 200 chromatography; the human breast cancer SKBr3 cell FAS was partially purified by ultra-centrifugation. The FAS was interacted with different concentrations of Triclosan with different times before catalyzing. Then the substrates of FAS were added to the reaction system. The inhibitory activities of triclosan against the FAS were investigated using spectrophotometric assays. RESULTS: In the goose uropygial gland group, FAS was purified as a single band at 250kDa with SDS-PAGE. The inhibitory activities of triclosan(12.5 micromol/L) at 0, 5, and 10 minute on FAS were 26.40%, 28.30%, and 43.93%, respectively. The inhibitory activities of triclosan (25.00 micromol/L) at 0, 5, and 10 minute on FAS were 46.22%, 50.28%, and 97.05%, respectively. The inhibitory activities of triclosan (100.00 micromol/L) at 0, 5, and 10 minute on FAS were 98.11%, 97.75%, and 97.37%, respectively. In human SKBr3 breast cancer cell group, the inhibitory activities of triclosan (25, 50, 100, and 200 micromol/L) at 5 minute on FAS were 20.00%, 26.67%, 60.00%, and 100%, respectively. CONCLUSION: Triclosan inhibits the FAS from goose uropygial glands and human breast cancer SKBr3 cells. The inhibitory activities depended on the concentrations of triclosan and the interaction times between triclosan and FAS before catalyzing. PMID- 12654185 TI - [An experimental research on inhibitory action of skeletal muscle conditioned medium on metastases of lung carcinomas]. AB - BACKGROUND & OBJECTIVE: Malignant tumors spread and metastasize in the majority of the organs, but are very rare in skeletal muscles. This study was conducted to explore the effect of organic microenvironment of skeletal muscles on the proliferation of pulmonary large cell carcinomas with different metastatic potential and to investigate the mechanism of the rarity of metastases in skeletal muscles. METHODS: Primary culture of newborn Wistar rat skeletal muscle cells was established, and the murine skeletal muscle conditioned medium(MMCM)was prepared to test its effect in vitro on pulmonary large cell carcinomas with different metastatic potential (PLA-801C with lower potential and PLA-801D with relatively higher potential) by MTT assay. Adriamycin was used as positive control for MMCM; murine benign renal cells BHK-21 were used as negative control for lung carcinoma cells. RESULTS: Proliferation of tumor cell lines of both PLA 801C and PLA-801D was significantly restrained when cultured with MMCM, while BHK 21 cells were not affected(P< 0.05). Compared with PLA-801C (significant only in primary MMCM), PLA-801D showed significantly decreased proliferation even when cultured in higher reciprocal of MMCM dilution(1/16 of primary MMCM). CONCLUSION: Skeletal muscle cells could selectively inhibit the proliferation of cancerous cells in vitro while benign cells are not affected. Tumor cells with higher metastatic potential are more sensitive to this effect. PMID- 12654186 TI - [Expression of cell cycle-regulatory proteins in squamous cell carcinoma of the esophagus]. AB - BACKGROUND & OBJECTIVE: Tumor cell cycle analysis has indicated that tumors with a higher proliferation rate showed more aggressive clinical behavior. Cell cycle regulatory proteins Ki-67, cyclin A, and p27 are associated with cell proliferation. The objective of this study was to observe the character of expression of cell cycle-regulatory proteins Ki-67, cyclin A, and p27 in esophageal carcinoma and to clarify the relationship between clinicopathologic and these proteins. METHODS: Immunohistochemistry for Ki-67, cyclin A, and p27 were carried out. Ki-67 and cyclin A staining index (SI), and p27 labeling index were examined and detailed pathologic examinations were conducted on tumors from 60 patients (48 males and 12 females with surgically resected esophageal squamous cell carcinoma). RESULTS: Ki-67, cyclin A, and p27 immunostaining were all confined to the nuclei in both neoplastic and non-neoplastic cells. The staining indexes(SIs) of Ki-67 and cyclin A, were significantly higher in poorly differentiated squamous cell carcinoma (SCC,27.2+/-4.9;15.4+/-5.3) than in well differentiated SCC(20.6+/-6.3;11.3+/-6.4,P< 0.05). The p27 positive immunostaining in the nuclei in well-differentiated SCC(36%)were higher than that in the other tumor types (29%;18%), but there was no significance(P< 0.25,P >0.05). CONCLUSION: The expression levels of Ki-67, cyclin A, and p27 may suggest the proliferative activity of cancer cells in Chinese patient with SCC of esophagus. The cell cycle-regulatory proteins Ki-67 and cyclin A over-expression correlates with poor SCC differentiation in patients with esophageal carcinoma. PMID- 12654187 TI - [Regularity and therapeutic strategy of cervical lymph node metastasis in squamous cell carcinoma of the tongue]. AB - BACKGROUND & OBJECTIVE: At present, whether neck dissection should be employed in squamous cell carcinoma (SCC) of the tongue with clinically node negative neck(cN0) is still controversial. This study was designed to explore the regularity of cervical lymph node metastasis in SCC of the mobile part of the tongue, and to discuss the theories and principles of employment of selective neck dissection in cN0 cases. METHODS: A retrospective research was performed on clinical data of 214 cases of SCC of the tongue treated with surgery from 1991 to 1997. Distribution of cervical lymph node metastasis of cN0 pN(+)= (pathologically node positive) and cN(+) (clinically node positive) pN(+)= cases were analyzed; the survival rates of different groups were compared; the factors that impact the survival of SCC of the tongue were screened out by Cox regression analysis. RESULTS: Cervical lymph node metastases were found in 69 cases. The metastatic rate was 32.2%. Metastases occurred in level I, II, III, IV, V of the ipsilateral neck were 22.3%, 33.5%, 22.3%, 4.6%, 1.0%, in level I,II,III, IV,V of the contralateral neck were 6.6%, 3.6%, 3.0%, 2.0%, 0.5%, respectively. The 5 year survival rates of the pN(+)= group and the pN0 group were 47% and 83%, respectively(P< 0.001). T stage and N stage were independent factors that impact the long-term outcome of SCC of the tongue. CONCLUSION: Level I,II,III of the ipsilateral neck tend to be involved when cervical lymph node metastases occur. Selective neck dissection can be used to treat the cervical metastasis in cN0 cases, as well as to evaluate the cervical lymph node status in order to determine whether comprehensive neck dissection should be employed. PMID- 12654188 TI - [Optimized methods of sentinel node localization in cN0 tongue carcinoma]. AB - BACKGROUND & OBJECTIVE: At present, there is no clinical examination and biochemical marker available to assess the lymph node status of tongue carcinoma with clinically negative neck (cN0) accurately. Therefore, the treatment of the cN0 neck in tongue carcinoma is of blind to some extent. Application of sentinel node(SN) biopsy may possibly bring about the individualized management of the cN0 neck of tongue carcinoma. This study was designed to assess whether lymph node status can be accurately evaluated by SN biopsy and to screen out the optimized sentinel node localization method. METHODS: Preoperative nuclear lymphoscintigraphy and intraoperative methylene blue dye mapping were performed in 24 cases of tongue carcinoma to reveal the efficacy of sentinel node biopsy in evaluating the lymph node status in cN0 tongue carcinoma. Shortcomings and advantages of lymphoscintigraphy, methylene blue dye mapping and combination of both methods were discussed. RESULTS: Sentinel nodes were detected in all cases with both methods. Sentinel nodes were found with metastasis in 4 cases. No non sentinel node was found with metastasis while sentinel nodes were pathologically negative. The detection rates were 3.5 sentinel nodes per case by lymphoscintigraphy, and 2.7 by methylene blue dye, while only 2.2 sentinel nodes by combined methods. CONCLUSION: Either lymphoscintigraphy or methylene blue dye mapping can be used to assess the cervical lymph node status of cN0 tongue carcinoma accurately. Combination with both methods is the most efficient and practicable method in sentinel node detection. PMID- 12654189 TI - [Quantitative magnetic resonance (MR) imaging of bone marrow in leukemia]. AB - BACKGROUND & OBJECTIVE: There were many studies on the relationship between magnetic resonance (MR) imaging of bone marrow and clinical laboratory variables of the patients with leukemia; however, few of them had separated lymphoid leukemia (LL) from myeloid leukemia (ML). The current study was designed to investigate the role of signal intensity ratio (SIR) of MR Imaging in the characteristic diagnosis and tumor burden evaluation in leukemia by separately measurement of spinal marrow SIR in LL and ML. METHODS: Spinal marrow in 20 LL patients and 10 ML patients in initial consultant were examined with MR imaging. The diagnosis of leukemia in all the patients was proved by iliac marrow cytological examination. MR imaging of spinal marrow was performed with 0.5T super-conducting system. T1-weighted imaging with spin-echotechniques and T2 weighted imaging with turbo spin-echo techniques were performed. The SIR of spinal marrow to spinal cord in leukemia was calculated on midline sagittal T1 weighted imaging. Meanwhile,peripheral blood routine examination and bone marrow cytological examination was performed. RESULTS: The SIR of spinal marrow in 20 LL patients and 10 ML patients were 0.72+/-0.11 and 0.73+/-0.11, respectively. There was no statistically significant difference between LL and ML(P=0.836). The SIR of spinal marrow in LL negatively correlated with the percentage of blast lymphocyte in marrow(r =-0.836,P= 0.000),while the SIR of spinal marrow in ML negatively correlated with the percentage of blast myelocyte in marrow (r= 0.673,P= 0.033). CONCLUSION: The SIR of spinal marrow is limited in the characteristic diagnosis of leukemia because of its inability to differential LL from ML. The SIR of spinal marrow can be used to evaluate tumor burden in leukemia. PMID- 12654191 TI - [Expression of growth hormone receptor in hepatocellular carcinoma and its significance]. AB - BACKGROUND & OBJECTIVE: Recombinant human growth hormone (rhGH) has been proved effective in clinic, such as promoting protein synthesis and decreasing the mortality. But there are still many arguments on whether it can be used in hepatocellular carcinoma (HCC) patients. rhGH cannot work except that it combines to its own receptor growth hormone receptor (GHR). The current study was designed to explore the expression of GHR in HCC tissues and to investigate the viability of rhGH for HCC treatment. METHODS: Radioreceptor assays were used to determine growth hormone receptor (GHR) in 40 HCC tissues; 6 normal liver tissues were used as control. Receptor binding capacity(RT) and affinity constant(Kd) of GHR were calculated by Scatchard's method; the relationship between RT of GHR in cancer and clinicopathologic factors were also analyzed. RESULTS: The growth hormone- specific singular binding site, namely GHR, was detected respectively in 35 HCC cases and control tissues. The RT and Kd of GHR were 18.5416+/-4.1686 fmol/mg protein and 0.6319+/-0.1978 nmol/L in tumor tissues, 39.5467+/-3.4770 fmol/mg protein and 0.6167+/-0.1007 nmol/L in control tissues, respectively. Compared with the normal liver tissue, RT of GHR in tumor was lower (P< 0.05) but Kd did not show any difference(P >0.05). The RT of the GHR was negatively relevant to the tumor size and disease stage, but was not associated with differentiation of tumor, ages of the patients or whether the patient suffered from cirrhosis at the same time. GHR was undetectable in 5 cases. CONCLUSION: This study showed that most of the HCC tissues express low levels of GHR. Before their functions are well understood, rhGH should be very carefully used in HCC patients. PMID- 12654190 TI - [Significance of detection of K-ras gene mutations and CA19-9 in serum for diagnosis of pancreatic carcinoma]. AB - BACKGROUND & OBJECTIVE: The early diagnosis of pancreatic carcinoma is difficult. The serum tumor markers such as CA19-9 have a relatively high sensitivity but with low specificity. The oncogene K-ras is frequently mutated in pancreatic carcinoma and with high specificity. The aim of this study was to assess the feasibility of detection of K-ras mutation combined with serum content of CA19-9 as an approach for diagnosis of pancreatic carcinoma. METHODS: Serum DNA was extracted from 39 patients with pancreatic carcinoma. Mutation enriched polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to determine codon 12 mutations of K-ras. Serum content of CA19-9 was determined by radioimmunoassay. In addition, the sera from 17 patients with other pancreatic diseases and 21 healthy individuals were also analyzed as control. RESULTS: K-ras gene mutations at codon 12 were detected in the sera of 71.79%(28/39) patients with pancreatic carcinoma and 11.76%(2/17) of patients with benign pancreatic tumors. The positive rates of CA19-9 were 71.79% and 41.18%, respectively. Parallel combined test increased the diagnostic sensitivity to 94.87%; and serial combined test increased the diagnostic specificity to 94.12%. Negative results of K-ras gene and CA19-9 were obtained in all sera from healthy controls. CONCLUSION: Combined detection of K-ras mutation and CA19-9 could increase the sensitivity and specificity in diagnosing pancreatic carcinoma, and would seem to be merited in clinic. PMID- 12654192 TI - [Significance of cell-free Epstein-Barr virus DNA in monitoring prognosis of nasopharyngeal carcinoma]. AB - BACKGROUND & OBJECTIVE: It has been reported that cell-free Epstein-Barr virus (EBV-DNA) in plasma was useful in diagnosing and monitoring nasopharyngeal carcinoma (NPC). The current study was designed to evaluate the significance of EBV-DNA in monitoring the prognosis of nasopharyngeal carcinoma and to compare with VCA/IgA and EA/IgA. METHODS: EBV-DNA, VCA/IgA, and EA/IgA levels in plasma were detected in different NPC patients after radiotherapy, including 30 distant metastasis patients, 22 locoregional recurrence patients, 24 remission individuals who had been followed up more than 2 years after treatment. EBV-DNA was detected using real-time quantitative PCR system;VCA/IgA and EA/IgA were tested using regular immunofluorescence. In cohort study, the indexes were tested in different radiation periods for the 20 new cases of nasopharyngeal carcinoma. RESULTS: The median plasma EBV-DNA concentration was 135,100 copies/ml (interquartile range: 5,525-1,003,750) in metastasis group, 20,500 copies/ml (interquartile range: 0-58,500) in locoregional recurrence group and 0 copies/ml (interquartile range: 0-0) in continuous remission group (P< 0.05). The levels of VCA/IgA and EA/IgA had no significant difference in different groups. The high level of EBV-DNA concentration in metastasis group was more than that in locoregional recurrence group. At the level of 1,000,000 copies/ml, EBV DNA indicated distant metastasis of NPC with a specificity of 100% and a sensitivity of 27.3%; however, the sensitivity was 0 copies/ml in locoregional recurrence group. For the 20 new patients, EBV DNA concentration gradually decreased in the radiation period, 32,050 copies/ml (interquartile range: 3,880-317,750) before radiation, 0 copies/ml (interquartile range: 0-14,375) when 40 Gy radiation dose and 0 copies/ml (interquartile range: 0-2940) when the radiation finished (P< 0.05). However, the levels of VCA/IgA and EA/IgA had no significant difference. CONCLUSION: The plasma cell-free EBV-DNA is more valuable than VCA/IgA and EA/IgA for monitoring the prognosis of NPC patients. PMID- 12654193 TI - [Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) expression in superficial transitional cell bladder carcinoma]. AB - BACKGROUND & OBJECTIVE: Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) can promote the proliferation of endothelial cell and angiogenesis. They are important materials in growth, invasion, and metastasis of tumor. There were few reports of their difference in single and multiple superficial transitional cell bladder carcinoma. Our objective was to study the expression of them in single and multiple superficial transitional cell bladder carcinoma and their clinical significance. METHODS: Immunohistochemical method was used to study the VEGF and bFGF expression in 60 cases of superficial transitional cell bladder carcinoma and in 10 cases of normal bladder tissue (as control). RESULTS: The expression levels of VEGF (55.6%), bFGF (50.0%), and coexpression level of VEGF and bFGF (50.0%) in the multiple superficial transitional cell bladder carcinoma were higher than that in single tumor. The recurrent rate in the patients with VEGF high expression (61.1%) was higher than that in the patients with VEGF low expression (16.7%). CONCLUSION: The expression levels of VEGF and bFGF were correlated with the biological behavior of superficial transitional cell bladder carcinoma. PMID- 12654194 TI - [Safety of an E1B deleted adenovirus administered intratumorally to patients with cancer]. AB - BACKGROUND & OBJECTIVE: In recent years, great development has been made in cancer therapeutics with replication-competent viruses (oncolytic viruses). Genetically modified adenovirus is one of promising viruses. The current study was designed to evaluate the safety of intratumoral H101, an E1B deleted adenovirus. METHODS: A total of 15 patients with malignant tumors were treated with H101 in a dose-escalation manner (from 5 x 10(7) VP to 1.5 x 10(12) VP per day for 5 consecutive days). Systemic examination was performed prior to injection. Physical signs (including weight, temperature, respiration,pulse, and blood pressure) and toxicity were observed and recorded carefully during treatment. Adenoviral DNA in plasma, urine, and swabs of oropharynx, swabs of the injection site from different time-point were determined by PCR. RESULTS: Evaluation of toxicity in all 15 cases was available. All patients tolerated well following intratumoral injection of H101. Dose limited toxicity (DLT) and serious adverse event (SAE) were not found in the course of treatment. Main side effects were injection site pain (60.0%) and fever (33.3%). Adenoviral DNA was undetectable in plasma, urine, and swabs of oropharynx, swabs of the injection site. CONCLUSION: This study showed that the toxicities of intratumoral injection of E1B deleted adenovirus (H101) were lower and tolerable. There is no evidence indicating the invasion. These results suggested that intratumorally injected H101 is safe. PMID- 12654195 TI - [Damaging nucleus centromedianus thalami for treatment of cancer pain in experimental and clinical aspects]. AB - BACKGROUND & OBJECTIVE: There is no perfect method to control cancer pain. It is reported that nucleus centromedianus thalami plays a crucial role in the analgesia of central nerve system. The authors conducted this study, based on rat experiments, together with the clinical treatment of more than 90 cases involving various cancer pains, to explore the pain-relieving effects after damaging nucleus centromedianus thalami. METHODS: Ten SD rats, whose nucleus centromedianus thalami were damaged by electrolysis, were chosen, and then measured the pain degree by applying electricity to stimulate the tails of the rats. Meanwhile, another 10 rats, whose nucleus centromedianus thalami were not damaged, were chosen as the control group, among whom the same operation procedure as the above mentioned was carried out. The range of pain scale of the rats was measured by the alteration of the electric intensity. A total of 90 cases of intractable cancer pain were treated, including 36 cases of lung cancer, 21 cases of nasopharyngeal carcinoma, 10 cases of intestinal cancer, 8 cases of cancer of pancreas, 8 cases of osteocarcinoma, 4 cases of carcinoma of kidney, 3 cases of hepatocarcinoma. The brain stereotactic technique was used to damage the nucleus centromedianus thalami with radiofrequency coagulation lesions. The 10 grade method recommended by WHO was used to rank pain degree. RESULTS: Pain scale of rats in the first group rose from 0.152+/-0.034 mA prior to the damage to 0.326+/-0.05 afterwards, with a significant difference (P< 0.001), while the pain scale of the control group dropped from 0.142+/-0.027 mA prior to the operation to 0.138+/-0.035 mA afterwards, with no remarkable difference (P > 0.05). To patients with cancer pain, the average pain grade in this study went above 7 scores, but dropped to 0-3 scores after operation, according to the 10 grade method by WHO. A life-long tracing observation indicted that cancer pain in 24 cases relapsed to varying degrees but below 5 scores, the rest of the patients were analgesic persistently, 3 cases among whom lasted for as long as 2 years. The incidence cases of operational complications were 15 of somnolence, 10 of urinary incontinence, 8 of divagation, and 3 of unilateral oculomotor paralysis. These complications released after symptomatic treatments. CONCLUSION: Nucleus centromedianus thalami damage is an effective way to relieve cancer pain, as well as the complications should be paid attention. PMID- 12654196 TI - [Clinical study of 39 cases of inferior postcricoid carcinoma]. AB - BACKGROUND & OBJECTIVE: There were many treatment strategies for inferior postcricoid carcinoma; however, the total survival rates were not so good till now. This study was designed to investigate the effective method for treatment of inferior postcricoid carcinoma. METHODS: Thirty-nine cases of inferior postcricoid carcinoma from 1970 to 1997 in Department of Head and Neck, Cancer Center, Sun Yat-sen University were surveyed. The treatment modalities were classified into two groups: simple radiotherapy and multimodality (surgery operation plus postoperational radiotherapy). The 3-year and 5-year survival rates of the two groups were analyzed. The different local lymph nodes metastasis was explored at the same time. RESULTS: Both the 3-year and 5-year survival rates of the simple radiotherapy group (35.2% and 17.6%) were lower than another group (59.1% and 45.5%) with significant difference (P< 0.05). Of 39 follow-up patients, 18 were found with neck lymph node metastasis. Contralateral or (and) bilateral lymph node metastasis were found in 72.7%(13/18) of the patients with neck lymph node metastasis. CONCLUSION: Multimodality is an effective method for inferior postcricoid carcinoma. It will increase the survival rate of the inferior postcricoid carcinoma by paying more attention to contralateral or (and) bilateral lymph node metastasis and dealing with it actively. PMID- 12654197 TI - [Expression of vascular endothelial growth factor and microvessel density in ovarian tumor]. AB - BACKGROUND & OBJECTIVE: Angiogenesis is a critical step in growth, invasion, and metastasis of solid tumors, and determination of microvessel of density (MVD) could be used to evaluate angiogenesis. Vascular endothelial growth factor (VEGF) is an important angiogenic factor, which involves in tumor growth. This study was designed to investigate the relationship between the expression of VEGF and MVD in borderline ovarian tumor. METHODS: SP immunohistochemistry and in situ hybridization were used to investigate the expression of VEGF and VEGF mRNA in 69 samples of borderline ovarian tumor (BOT), 18 samples of benign tumor and 27 samples of ovarian carcinoma (OC). MVD was counted by endothelial cells immunostained with anti-factor VIII monoclonal antibody. RESULTS: The expression levels of VEGF protein and VEGF mRNA were middle in borderline ovarian tumors between in benign tumors and in carcinoma, with significant differences (P< 0.05). The expression of VEGF and VEGF mRNA were closely associated with clinic stage and MVD(P< 0.05), but were not associated with tissue type and peritoneal implants(P > 0.05). CONCLUSION: VEGF is associated with the development, invasion, and metastasis of ovarian tumor. VEGF and MVD were predictors for the prognosis of borderline ovarian tumor. PMID- 12654198 TI - [Interactive pathway of ARF-mdm2-p53]. AB - The inhibitor of cyclin-dependent kinase 4-p16INK4 alpha and its alterative reading frame (ARF)-INK4/ARF gene locates at CDKN2A locus of human chromosome 9p21. This locus encodes two overlapped genes: ARF gene and p16INK4 alpha. The amino acid sequences of two genes are completely different because they are encoded by alternative reading frames. ARF participates in the regulation of mdm2 p53 pathway by mdm2. Recent studies showed that ARF gene may play a role in tumorigenesis; the ARF gene promoter hypermethylation may be the principal mechanism in the inactivation of this gene. Here is a review of ARF-mdm2-p53 interacting pathway. PMID- 12654199 TI - [BRCA1 and genomic stability]. AB - BRCA1 is a 220kDa nuclear protein with multiple functional domains. It interacts directly or indirectly with a variety of important proteins, including oncogene proteins (c-myc, E2F), tumor suppressor proteins (p53, RB, BRCA2), DNA damage repair proteins (RAD50, RAD51), cell-cycle regulators (cyclin, CDK), transcriptional regulators (RNA polymerase II) and others related to the important biological events. BRCA1 is likely to play an important role in the maintenance of genomic stability through its activities in cell-cycle progression, DNA damage repair, transcriptional regulation, and apoptosis. Here, the authors provided a review of the biochemistry structure of BRCA1 as well as its role in maintaining the genomic stability. PMID- 12654200 TI - Effect of cervical instability in sympathetic cervical spondylosis. AB - OBJECTIVE: To investigate the etiology and treatment of sympathetic cervical spondylosis (SCS). METHODS: Twenty patients who with SCS had undergone operations for sympathetic cervical spondylosis were reviewed retrospectively from 1988 to 2000. Lateral views in flexion and extension of pre- and postoperative cervical X ray were analyzed to quantify cervical instability. RESULTS: Cervical instability was detected at one level in seven patients, two levels in ten patients, three levels in three. Cervical instability mainly occurred at C(3)-C(4) and C(4)-C(5). Cervical epidural block had a short time effect in the greater part of patients. Cervical discectomy and fusion at unstable segment was carried out in all 20 cases. The effective rate was 90%. CONCLUSIONS: Cervical instability at C(3)-C(4) or C(4)-C(5) maybe an important factor in the etiology of sympathetic cervical spondylosis. Cervical epidural block may provide diagnostic information. Anterior cervical fusion were effective to treat sympathetic cervical spondylosis. PMID- 12654201 TI - [Experience with transluminal stent-assisted angioplasty for intracranial vascular diseases]. AB - OBJECTIVE: To assess the value of stent-assistant angioplasty for intracranial vascular diseases. METHODS: Thirteen patients with intracranial vascular diseases were treated consecutively by stent-assistant angioplasty for different purposes. Of these patients 7 had symptomatic intracranial artery stenosis, 3 intracranial wide-neck aneurysms, 2 intracranial pseudoaneurysms, and 1 bilateral transverse sinus stenosis. Clinical procedures and technical data of the patients were retrospectively analyzed. RESULTS: In the 7 patients, the degree of intracranial artery stenosis reduced from 83% to 5%. Three months after the operation they exhibited good patency of stented lesions. Intracranial wide neck aneurysms (with 2 pseudoaneurysms) were successfully embolized with GDC in 5 patients. The patients with bilateral transverse sinus stenosis were implanted a self-expanded stent in the right transverse sinus. One patient failed because of the tortuous configuration of the left jugular bulb, but the patient's symptoms were improved quickly after the procedure. CONCLUSION: Stent-assistant angioplasty is a useful technique for selected intracranial vascular diseases, and it needs further investigation. PMID- 12654202 TI - [Endovascular thrombolysis and stent angioplasty for obliteration in cerebral venous sinuses]. AB - OBJECTIVE: To summarize the experience of treating the obliteration of the cerebral venous sinus in 17 patients by using direct thrombolysis and stent angioplasty. METHODS: All 17 patients with thrombosis and stenosis of the venous sinus were confirmed by digital subtraction angiography. Of these 3 patients had thrombosis in a single sinus and 14 had thrombosis in multiple sinuses. The circulating time was prolonged for over 13 seconds. The micro-catheter was preserved in the sinus for 5 days, followed by infusion of urokinase 1.5 million units and oral warfarin 3-5 mg each day. Stent angioplasty was done for 4 patients with obvious stenosis of the venous sinus detected by DSA after thrombosis. RESULTS: After contact thrombosis and stent angioplasty in sinuses of the 17 patients, remarkable recanalization of obliterated sinuses was achieved. After treatment, intracranial hypertension pressure (ICP) returned to normal in 7 patients, and 8 thrombosis relapsed in different degree after 7 days. Anticoagulation was prescribed. Only 2 patients showed the ICP above 280 mm H(2)O. No obvious relapse was found in 15 patients during the follow-up for 3-60 months. CONCLUSION: Our results demonstrated that successive thrombolysis and stent angioplasty for occlusion and thrombosis of the venous sinus are effective in promoting drainage of cerebral venous blood and rapidly decreasing ICP. PMID- 12654204 TI - [Laparoscopic total mesorectal excision for low or ultralow anterior resection of rectal cancer with anal sphincter preservation]. AB - OBJECTIVE: To assess the feasibility of laparoscopic total mesorectal excision (TME) for low or ultralow anterior resection of rectal cancer. METHODS: Excision of the mesorectum and low (ultralow) colo-anal anastomoses were performed laparoscopically in 62 patients with low rectal cancer based on the concept of TME and double stapling technique (DST). RESULTS: Sixty-two operations with TME and DST were performed in a totally laparoscopic manner, and only one was converted to open procedures because of dysfunction of coagulation. The operative time was 125 min (110-210 min) and the operative blood loss 20 ml (5-80 ml). The time for bowel function recovery and post-operatively dietary intake was 1-2 days. Twenty-eight patients received postoperative analgesics. Average hospital stay was 8 days (5-14 days). Complications were observed in only 2 of the 62 patients, one had suffered from urinary retention and the other, anastomotic leakage. CONCLUSIONS: Totally laparoscopic excision of the mesorectum for low or ultralow anterior resection of rectal cancer is a minimally invasive technique with sphincter preservation, less postoperative pain, and rapid recovery. PMID- 12654203 TI - [Initial application of brain protection device in dilatation and stenting of carotid and vertebral artery stenosis]. AB - OBJECTIVE: To report the initial application of brain protection device in the dilatation and stenting of atherosclerotic stenosis of the carotid and vertebral artery. METHODS: Eighteen patients with 21 atherosclerotic stenoses of the carotid or vertebral artery underwent dilatation and/or stenting with brain protection device (filterwire or angioguard). The clinical results were summarized and the indispensability and feasibility of the device was discussed. RESULTS: With the aid of brain protection device, endovascular dilatation and (or) stenting were performed in all the patients with 21 stenoses of the carotid or vertebral artery. The interventional manipulation was successful and no complications occurred. CONCLUSIONS: Brain protection device is helpful to decrease the embolic complication caused by atherosclerotic plaque and thromboembolus and to increase the security of interventional therapy, during the dilatation and/or stenting of stenosis of the carotid or vertebral artery. PMID- 12654205 TI - [Multiple primary malignant neoplasms]. AB - OBJECTIVE: To investigate the epidemiological and clinical features of multiple primary malignant neoplasms (MPMNs). METHODS: One hundred and twenty-five cases of more than or equal to three primary malignant neoplasms detected in the period of 1958 to 2001 were studied retrospectively in terms of frequency, age of onset, sex ratio, tumor location and the interval between sequential tumors. RESULTS: The overall incidence of MPMNs was 0.090%; for three, four and five MPMNs was 0.078%, 0.009% and 0.001%. The mean age of onset of MPMNs was 64 years. The sex ratio, of male to female, was 1.0:1.2. The mean time interval was 7.1 years, and it reduced from 5.7 to 2.4 years in a descending order of the first, second, third, forth and fifth primary tumors. Three, four and five primary malignant neoplasms were predicated respectively in the large intestine, breast, lung, esophagus, uterus, skin; the large intestine, breast, skin stomach, urinary bladder, uterus; and the large intestine, breast, skin, parotid glands. CONCLUSIONS: The overall incidence of multiple primary malignant neoplasms seems to ascend. The age of onset of MPMNs tends to be increase. The incidence of in women is higher than in men. The time interval from the first to fifth primary tumor is decreasing. The susceptible organs are identical organ, bigeminal organs or organs of the same system. PMID- 12654206 TI - [Comparison of colonic J-pouch and straight coloanal anastomosis after low anterior resection for rectal carcinoma: a meta-analysis of 8 randomized trails]. AB - OBJECTIVE: To evaluate the effect of colonic J-pouch coloanal anastomosis after low anterior resection for mid or low rectal cancer on improving defecation and anorectal physiology. METHODS: To make a meta-analysis, prospective randomized controlled trial of with or without colonic J-pouch after low anterior resection for mid or low rectal cancer was conducted. The key words included rectal cancer, J-pouch (or J-pouch) and those randomized from selected reports. The data on bowel function and physiological function of the anal canal and (neo) rectum were meta-analyzed using fixed effect model and random effect model. RESULTS: Eight randomized trails including 378 patients entered this study. After one year follow-up, the functional results showed that there were significant differences in stool frequency per day, urgency and use of medication between colonic J-pouch group and straight coloanal anastomosis group. On physiological function of the anal canal and (neo) rectum, there was significant difference only in rectal compliance between the two groups. But there was no significant difference in other 4 items. CONCLUSIONS: The functional improvement gained from colonic J pouch coloanal anastomosis continues to benefit the patient with mid or low rectal cancer for at least 12 months. It is necessary to further study about the effect of colonic J-pouch coloanal anastomosis on physiological function of the anal canal and (neo) rectum. PMID- 12654208 TI - [Arthroscopy for the diagnosis and treatment of hip joint disease]. AB - OBJECTIVE: To evaluate the value of arthroscopy for the diagnosis and treatment of hip joint disease. METHODS: Arthroscopy was performed in 54 patients (68 hips) from July 1998 to September 2001. Of the 54 patients, 36 were male and 18 female, aged from 17 to 64 years. The disease was located on the left side in 18 patients, the right side in 22, and both sides in 14. In this group, osteoarthritis was noted in 21 patients, avascular necrosis of the femoral head in 16, infection of the hip joint in 3, chondrosarcoma in 2, synovial chondromatosis in 7, acetabular labral tears in 2, and spondylitis in 3. The procedures of arthroscopy consisted of removal of loose bodies, debridement, and biopsy. RESULTS: The operation made accurate diagnosis possible for chondrosarcoma, acetabular labral tears, and infection of the hip joint. After debridement of the hip joint for avascular necrosis of the femoral head, osteoarthritis and spondylitis, most patients in this group were free from pain, function and range of the motion were restored and the pathological process was delayed. The Harris hip scores increased by 27 on average. CONCLUSION: Arthroscopy plays an important role in accurate diagnosis and treatment of hip joint. PMID- 12654207 TI - [Screening and cloning of genes related to varicose great saphenous vein accompanying with primary deep vein valve insufficiency]. AB - OBJECTIVE: To screen the genes related to the occurrence and development of varicosis of the great saphenous vein in the patients with primary deep vein valve insufficiency. METHODS: Using mRNA fluorescent differential display reverse transcriptive polymerase chain reaction (FDD-RTPCR), different genes expressed in the varicose great saphenous veins in patients with primary deep vein valve insufficiency and corresponding normal human tissues were compared. Differentially expressed cDNA fragments confirmed by Northern blot were compared and then cloned into the pGEM-Teasy vector. Positive clones were selected and sequenced. All the sequences were put into GenBank and analyzed by BLASTN software to search for their genetic origins. RESULTS: Altogether 37 different cDNA fragments were obtained and 30 of which were confirmed by Northern blot. Analysis of the sequences by BLASTN software showed that C(610) fragment (NO. 18 cDNA clone) shared 96% homology with the mRNA sequence of the human Mckusick Kaufman syndrome gene (MKKS gene). CONCLUSION: C(610) fragment is highly homologous with the mRNA sequence of the human MKKS gene and is closely related to the development of varicosis of the great saphenous vein in patients with primary deep vein valve insufficiency. PMID- 12654209 TI - [Treatment of simple renal cysts with small incision]. AB - OBJECTIVE: To analyze the operative characteristics and indications small incision for treating simple renal cysts. METHODS: 19 patients with simple renal cyst were treated by ablation with small incision from January. 1995 to September. 2001 at our institute. Surgical technique, curative effect and indications were analyzed. RESULTS: In the 19 patients the operative procedure took 34 +/- 15 min (20-80 min) on average and the mean hospitalization was 4.4 +/ 1.5 d (3-8 d). Neither complication nor recurrence occurred from 3 to 6 months during the follow-up. CONCLUSIONS: Ablation with small incision is significantly advantageous for operation, recovery, short hospital stay, and lower cost. This operation is indicated for simple renal cyst in the lower, middle pole of the kidney. PMID- 12654210 TI - [Patella ligament and femoral quadriceps tendon rupture]. AB - OBJECTIVE: To study the diagnosis and treatment of rupture of patella ligament and Femoral quadriceps tendon. METHODS: Twenty-three patients with rupture of the patella ligament and femoral quadriceps tendon were treated from March 1990 to August 2000. They were retrospectively evaluated with regard to patient's age, cause of injuries, characteristics of injuries, symptoms and signs, supplemental checkup, patella position, operative method and time interval from injury to treatment, immobilization, motion of knee, functional recovery etc. RESULTS: They were followed up for 6 years (4 months-11 years) on average. Thirteen of 15 acute injuries after treatment showed excellent results and good results in 2. In 5 old injuries of the extensor mechanism after treatment 2 got excellent results (3 lost to follow-up). CONCLUSION: Excellent or good results could obtain in patients with acute injuries after operation. The results of old injuries are not as good as acute injures, but better results could also achieve after repair. PMID- 12654211 TI - [Operative stress response and energy metabolism after laparoscopic cholecystectomy and open cholecystectomy]. AB - OBJECTIVE: To determine the level of neurohormonal operative stress response reactive protein (CRP) and rest energy expenditure (REE) after laparoscopic cholecystectomy (LC) and open cholecystectomy (OC). METHODS: Twenty-six consecutive patients with noncomplicated gallstones were randomized for LC (n = 14) and OC (n = 12). Plasma concentrations of somatotropin, insulin, cortisol and CRP were measured. The levels of REE were also measured. RESULTS: On the third postoperative day, the insulin levels were lower than those before operation (P < 0.05). On the first postoperative day, the levels of somatotropin and cortisol were higher in OC than in LC. After operation, the parameters of somatotropin, CRP and cortisol increased compared to the preoperative period in all patients (P < 0.05). On the all-postoperative day, the CRP levels were higher in OC than in LC (P < 0.05). After operation, the REE level increased in OC and LC (P < 0.05). On the all-postoperative day, the REE levels were higher in OC than in LC (P < 0.05). CONCLUSIONS: LC results in less prominent stress response and smaller metabolic interference compared to open surgery. These benefit the restoration of stress hormones, nitrogen balance, and energy metabolism. However, LC can also induce acidemia and pulmonary hypoperfusion because of pneumoperitoneum during surgery. PMID- 12654213 TI - [Comparison of perioperative myocardial injury between off-pump coronary artery bypass grafting and conventional coronary artery bypass grafting]. AB - OBJECTIVE: To compare the perioperative release levels of cardiac troponin I (cTnI) between off-pump coronary artery bypass grafting (OPCAB) or conventional coronary artery bypass grafting (CCABG) in an attempt to detect myocardial injury. METHODS: Fifty-nine patients with instable angina underwent coronary artery bypass grafting by OPCAB in 34 patients aged (59.15 +/- 1.71) years or CCABG in 25 patients, aged (54.46 +/- 1.81) years. RESULTS: Baseline characteristics were similar. The number of grafts was similar (OPCAB, mean 2.90; CCABG, mean 3.2), and no patient died. Postoperative myocardial serum enzyme measures were significantly lower in OPCAB, suggesting less myocardial injury. OPCAB patients did not receive blood transfusion, and had higher hematocrit at discharge. Most of OPCAB patients were extubated in 4 hours. CONCLUSIONS: Compared with CCABG, OPCAB may achieve similar outcomes; it reduces transfusion volume and creates less myocardial injury. PMID- 12654212 TI - [Primary common bile duct closure after choledochotomy]. AB - OBJECTIVE: To investigate the rationality and feasibility of primary closure of the common bile duct after choledochotomy for common bile duct calculi. METHODS: From January 1990 to June 2001, 386 patients with the evidence of stones in the common bile duct underwent choledochotomy. Among them, 215 received primary closure of the common bile duct (group A) and 171 T-tube drainage (group B). The patients with emergency operations were excluded. Intraoperative choledochoscopy or cholangiography was routinely performed to rule out the possibility of retained stones. The duct was meticulously stitched using 0/3 to 0/5 absorbent sutures for primary closure. A T-tube was placed in the subhepatic space in the patients of both groups. RESULTS: Postoperative bile leakage was seen in 9 patients of group A and in 5 of group B, respectively (P > 0.05), and no reoperations were necessary. After surgery, the average time and volume of transfusion was 4.9 days and 9.1 liters in group A, versus 7.3 days and 12.8 liters in group B (P < 0.01). The patients in group B had a longer postoperative hospital stay than the those in group A (average 17.6:10.0 days, P < 0.01). T tube removal resulted in bile peritonitis in 5 patients at day 16, 17, 19, 21 and 22 after surgery in group B, and 3 patients required repeated surgery. CONCLUSIONS: Primary closure of the common bile duct after choledochotomy is safe, effective, and inexpensive in selected patients with common bile duct calculi, and should be regarded as an alternative procedure. PMID- 12654214 TI - [Preparation and osteogenic activity of combining calcium phosphate cement with bone morphogenetic protein composite as a bone graft material]. AB - OBJECTIVES: To construct new type of bone graft material by combining calcium phosphate cement (CPC) with bone morphogenetic protein (BMP), and then to detect its osteogenic activity. METHODS: The surface of CPC and CPC/BMP composite were observed by scanning electron microscope (SEM). CPC and CPC/BMP pellets were separately implanted into the thigh muscle pouches of mice. Samples obtained at different times were tested by histological analysis, SEM, organic substance detection, and alkaline phosphatase (ALP) measurement to observe the induced ectopic bone formation. RESULTS: Under SEM, the CPC and CPC/BMP composite was found to consist primarily of platy crystals, granular crystals and some small rod-like crystals with micropores about 10-50 microm in size. BMP about 1-5 microm in size was seen like micro globules distributing evenly in the micropores. Newly formed cartilage or bone was not found in the CPC group. In the CPC/BMP group, mesenchymal cells were proliferated and abundant cartilage was found in one week. Woven bone appeared at 2 weeks. New bone formation increased with bone marrow at 4 weeks. At 8 weeks, the implanted CPC/BMP became heterogeneous and a lot of collapsed granules were observed. At the end of 16 weeks, mature lamellar bone appeared and the volume of the implanted CPC/BMP became smaller. One week after implantation, the ALP increased evidently in the CPC/BMP groups and reached the highest level at the 4th week, which was about 168 U/L. The content of organic substance in specimens increased from 22% to 39% by the end of the 16th week, showing the continuous calcification and formation of new bone. SEM also showed that the CPC/BMP composite had good potentiality of ectopic bone induction, and the new bone formed accompanied by the slow degradation of the material. CONCLUSION: The results of this study suggested that the CPC/BMP composite could be used as material for bone graft substitute. PMID- 12654215 TI - [Influence of warm ischemia injury on energy metabolism and survival of liver graft in rats]. AB - OBJECTIVES: To investigate the energy metabolism and post transplantation survival of liver graft under different warm ischemia times (WIT) in rats and determine the maximum limitation of liver graft to warm ischemia. METHODS: According to WIT, the rats were randomized into 7 groups, and WIT were 0, 10, 15, 20, 30, 45, 60 minutes respectively. The indexes of energy metabolism were measured by reversed-phase high performance liquid chromatography (HPLC) and all liver graft specimens were subjected to ultrastructural observation. After orthotopic liver transplantation (OLTx), the recovery of energy metabolism of liver graft after 24, 48 hours and the rats' survival were observed. RESULTS: The levels of adenosine triphosphate (ATP) and energy charge (EC) decreased gradually after different WIT in a time-dependent manner, and especially significant within 30 minutes. The levels of ATP and EC of liver grafts were largely recovered after 24 hours of OLT within 30 minutes of warm ischemia, partially recovered after 48 hours of OLT with 45 minutes of warm ischemia and hardly recovered even after 48 hours of OLT with 60 minutes of warm ischemia. The rat survival time after OLT was not significantly different within 30 minutes of WIT, while the long-term survival was insulted with 45 and 60 minutes of WIT. CONCLUSIONS: The levels of ATP and EC after OLT may be the important criteria to evaluate the quality of liver graft. WIT of liver graft is closely related to both the recovery of hepatic energy metabolism and the liver graft survival. PMID- 12654216 TI - [Growth hormone insensitivity of rats under the endotoxemic condition]. AB - OBJECTIVE: To investigate the mechanism of growth hormone insensitivity of rats under the endotoxemic condition. METHODS: Sprague Dawley rats (n = 180) were injected endotoxin, TNF-alpha, and IL-6 respectively. Part of endotoxin injected rats were treated with exogenous somatotropin simultaneously, and all rats were killed at different time points. Liver expression of IGF I, GHR and SOCS-3 mRNA was detected by RT-PCR, the levels of growth hormone (GH) were measured by radioimmunoassay, and the levels of TNF-alpha and IL-6 were detected by ELISA. RESULTS: Serum GH levels showed no significant change after endotoxin injection; however, liver IGF I and GHR mRNA expressions were obviously down-regulated in endotoxemic rats, with the lowest decrease of 53% and 89% respectively. Although SOCS-3 mRNA was weakly expressed in control rats, it was strongly up-regulated in endotoxemic rats and the marked increase was 7.84 folds. The higher LPS dosage induced marked GHR mRNA down-regulation and marked SOCS-3 mRNA up-regulation. Exogenous GH made IGFI mRNA expression increase 25% in the control rats, but it did fail to prevent the decline in IGFI mRNA expression in endotoxemic rats. Endotoxin stimulated the production of TNF-alpha and IL-6, and the elevated IL-6 levels showed a positive correlation with increased SOCS-3 mRNA expression. Liver GHR mRNA expression was obviously down-regulated after TNF-alpha i.v. injection, but for IL-6, it mainly up-regulated the liver SOCS-3 mRNA expression. CONCLUSION: The growth hormone insensitivity could be induced by LPS injection, which might be associated with down-regulated GHR mRNA expression and up regulated SOCS-3 mRNA expression. The in vivo biological activities of LPS may be partially mediated by TNF-alpha and IL-6 at different aspects. PMID- 12654217 TI - [A clinical study on laser peripheral iridoplasty for primary angle-closure glaucoma with positive provocative tests after iridectomy]. AB - OBJECTIVE: To evaluate the therapeutic effects of laser peripheral iridoplasty for primary angle closure glaucoma with positive dark room and prone test after laser peripheral iridectomy. METHODS: A long-term prospective study of 56 eyes (34 cases) with primary angle-closure glaucoma was carried out. The patients presented with positive dark room and prone provocative test after laser peripheral iridectomy, and laser peripheral iridoplasty by Double-Frequency Nd:YAG laser was performed on them. Their extent of goniosynechia was less than 1/2 circumference of the anterior chamber angle. Forty-nine eyes (27 cases) of all those studied were acute angle-closure glaucoma and the other 7 eyes (7 cases) were chronic angle-closure glaucoma. The inferior peripheral anterior chamber depth, anterior chamber angle, the configuration of peripheral iris and intraocular pressure were observed carefully, and the dark room, prone and mydriatic provocative tests were performed. The postoperative follow-up ranged from 1 year to 4 years. RESULTS: The results showed that in all these cases, the peripheral anterior chamber depth was increased, the anterior chamber angle was widened on goniscope and the trabecular meshwork could be visualized widely in static state. All patients did not have ocular hypertension and damage of visual field in the follow up. Mydriatic, dark room and prone provocative tests following laser peripheral iridoplasty were negative. CONCLUSIONS: In some acute angle-closure glaucoma and chronic angle-closure glaucoma patients, the pupillary block is relieved by laser peripheral iridectomy, but provocative tests can also be positive because of the abnormal configuration of the peripheral iris. Pupillary dilation results in peripheral iris bunching, and the surface of the trabecular meshwork can be blocked, hypertension then occurs. But the laser peripheral iridoplasty can improve the shape of the peripheral iris effectively and it can prevent the disease from deteriorating. The goniscopic examination and provocative tests following laser peripheral iridectomy are very important and also effective in detecting this kind of glaucoma. PMID- 12654218 TI - [Drug therapy for residual angle-closure glaucoma after laser iridectomy]. AB - OBJECTIVE: To observe and compare the effect of 0.005% latanoprost and 0.5% timolol solutions in the treatment of post-iridectomy residual angle-closure glaucoma. METHODS: Laser peripheral iridectomy was performed and 68 cases (68 eyes) of post-iridectomy residual angle-closure glaucoma were selected and treated with 0.005% latanoprost once at night as the study group and 0.5% timolol twice a day as the control group Intraocular pressure (IOP) was measured before treatment, on day 3, week 1, week 2, week 4, month 2, month 3 and month 6 in the follow-up, and visual acuity, visual field and C/D ratio were monitored. RESULTS: Of 68 patients, 56 (82.4%) complete the 6 month follow-up. The basic IOP for all subjects in two groups before treatment was not statistically different (t = 0.236, P = 0.814). After the treatment for three days, the efficiency in IOP reduction was statistically different between 2 groups with repeated measure ANOVA (F = 287.441, P < 0.001). In each group, the IOP after treatment was statistically different from that at baseline, with t = 14.103, P < 0.001 and t = 10.308, P < 0.001 in the latanoprost and timolol groups respectively. For those patients who finished 6 months' follow-up, the IOP difference between the 2 groups was statistically significant with repeated measure ANOVA (F = 74.259, P < 0.001). The IOP was statistically reduced in both the latanoprost and the timolol groups with F = 148.787, P < 0.001 and F = 54.875, P < 0.001 respectively. At each follow-up time, the IOP in latanoprost group was statistically different from that of the timolol group with t values to be -6.127, -5.930, -6.201, 5.931, -7.412, -6.640 and -6.756 (P < 0.001). The IOP was statistically different from that before treatment in each group with a mean difference to be -7.034 approximately -6.897 in the latanoprost group and -4.704 approximately -5.000 in the timolol group (P < 0.001). The IOP was not statistically different within each group in the follow-up with mean difference of 0.034 approximately 0.138 (P = 0.904 approximately 0.629) in the latanoprost group and a mean difference of 0.037 approximately 0.296 (P = 0.910 approximately 0.366) in the timolol group. The best-corrected visual acuity, visual field and C/D ratio were not significantly changed during follow-up. CONCLUSIONS: 0.005% latanoprost and 0.5% timolol can be used in the drug therapy of post-iridectomy angle-closure glaucoma and the efficiency of 0.005% latanoprost is better than of 0.5% timolol. Drug therapy of post-iridectomy angle-closure glaucoma is a practical strategy. PMID- 12654219 TI - [Frequency-doubling perimetry in examination of open-angle glaucoma with hemifield defect]. AB - OBJECTIVE: To determine whether frequency-doubling perimetry (FDP) can find visual field defect of early glaucoma which can not be found by conventional perimetry. METHODS: Forty-nine eyes of 49 patients with open-angle glaucoma and hemifield defect detected by Humphery Field Analyzer (HFA) were tested by both FDP (threshold test C-20) and Heidelberg Retina Tomograph (HRT). For each hemifield, its FDP result was evaluated. According to the FDP results in intact HFA hemifields, the patients were divided into FDP normal group and FDP abnormal group. The HRT parameters of half disk corresponding to the intact HFA hemifields were compared between these two groups. RESULTS: There was a strong correlation between the mean sensitivity of FDP and mean deviation of HFA results (r = 0.58, P < 0.01). In intact HFA hemifields, 41% (20/49) of eyes showed visual loss in FDP tests, while in defective HFA hemifields, only 2% (1/49) of eyes was normal in FDP test. For the HRT parameters of half disk corresponding to the intact HFA hemifields, cup area and cup/disk area ratio were statistically bigger and rim area and rim volume were statistically smaller in FDP abnormal group than in FDP normal group. CONCLUSION: FDP is able to detect early glaucomatous visual field damage which can not be detected by HFA, but can be explained by the optic nerve glaucomatous changes in HRT parameters. PMID- 12654220 TI - [Effects of pilocarpine in new ophthalmic formulations on intraocular pressure in rabbits with ocular hypertension]. AB - OBJECTIVE: To assess the effects of pilocarpine (PC) incorporated into an in situ gelling vehicle, submicron emulsion vehicle, polyvinyl alcohol vehicle and PC solution on intraocular pressure (IOP) in rabbits with carbomer-induced elevation of IOP. METHODS: Forty-three rabbit eyes with carbomer-induced elevation of IOP were randomly divided into 5 groups. Each eye in every group received topical application of PC from one of the ophthalmic formulations. IOP was measured before and 1, 2, 3, 4, 5, 7, 10, 12, 22 and 24 hours (h) after topical ocular administration of the ophthalmic formulations. RESULTS: The reduction in IOP lasted for 7 h and the maximum decrease was (12.25 +/- 5.28) mm Hg in the hypertensive eyes treated by 2% PC solution. The reduction in IOP lasted for 12 h and the maximum decrease was (13.67 +/- 4.61) mm Hg in the hypertensive eyes treated by 2% PC gel. The reduction in IOP lasted for 24 h and the maximum decrease was (13.63 +/- 3.58) mm Hg in the hypertensive eyes treated by 4% PC gel. The reduction in IOP lasted for 12 h and the maximum decrease was (13.55 +/- 3.90) mm Hg in the hypertensive eyes treated by 2% PC submicron emulsion. The reduction in IOP lasted for 10 h and the maximum decrease was (10.89 +/- 4.04) mm Hg in the hypertensive eyes treated by 2% PC polyvinyl alcohol. The duration of action for lowering IOP lasted longer following administration with new ophthalmic formulations as compared to 2% PC solution. The series of prolongation is as follows: 4% PC gel > 2% PC gel = 2% PC submicron emulsion > 2% PC polyvinyl alcohol > 2% PC solution. The maximum reduction in IOP was not significantly different among the groups. No clinical evidence of ocular toxicity occurred in all groups except mild conjunctival injection. CONCLUSION: These results indicate that the duration of action for lowering IOP is longer in PC gel, PC submicron emulsion and PC polyvinyl alcohol as compared to PC solution. The duration of action with 4% PC gel lasts the longest among all the groups. A single dose of 4% PC gel applied once daily can reduce IOP for 24 h. PMID- 12654221 TI - [Non-penetrating trabecular surgery with application of mitomycin C]. AB - OBJECTIVE: To evaluate the clinical effect of non-penetrating trabecular surgery (NPTS) with mitomycin C (MMC) in primary open-angle glaucoma (POAG). METHODS: Twenty-five eyes of 18 patients with POAG underwent NPTS with MMC during the operation. The mean follow-up was 16.6 months (6 - 28 months). RESULTS: The mean preoperative intraocular pressure (IOP) was (27.93 +/- 7.62) mm Hg (1 mm Hg = 0.133 kPa) which decreased to a mean postoperative IOP of (14.62 +/- 3.53) mm Hg (P < 0.001). After operation, there was an obvious diffuse bleb in each eye, in the 3 months after operation, there were functional blebs in 21 eyes, and the successful filtration was ceased in 4 eyes. The anterior chamber reaction was mild. Hyphema occurred in 1 eye after operation. Intra-operatively, in 4 eyes microperforations occurred in the trabecular meshwork without iris prolapse, and after operation, under the gonioscope, the perforation could be observed in these eyes. In another 2 eyes, there was no intra-operative perforation, but a small fissure was seen by gonioscopy. CONCLUSIONS: NPTS with application of MMC can lower the IOP effectively in this group of patients. The recovery of vision after operation is quick. A few complications occurred after operation. It is a promising technique to treat POAG. PMID- 12654222 TI - [Chop technique with lens loop-pad and foldable intraocular lens implantation]. AB - OBJECTIVE: To evaluate the use of chop technique with lens loop-pad and foldable intraocular lens implantation for cataract surgery. METHOD: Chop technique with self-designed lens loop-pad and foldable intraocular lens implantation were performed on 73 eyes of 64 cases with senile cataract. RESULTS: The intraoperative complication was posterior capsular rupture in only 2 cases. All the patients had very mild postoperative reaction and round pupil. The corrected visual acuity was improved to 0.5 or over in 79.5% (58 eyes) at one week postoperatively, and in 89.0% (65 eyes) at three months postoperatively. The mean percentage of endothelial cell loss was 14.1% (t = 17.46, P < 0.001) at the superior area of cornea and 12.2% (t = 17.52, P < 0.001) at the center of cornea. CONCLUSION: It is suggested that the chop technique with lens loop-pad simplify the procedure, and be safer and more reliable, achieving the goals of small incision cataract surgery. PMID- 12654223 TI - [Effects of laser in situ keratomileusis (LASIK) retreatment on the central corneal endothelium and retina and optic nerve]. AB - OBJECTIVE: To investigate the effects of laser in situ keratomileusis (LASIK) on the corneal endothelium, retina and optic nerve for correcting residual myopia caused by undercorrection or regression. METHODS: The pre- and post-operative values of central corneal endothelium and electrophysiological functions of retina and optic nerve were analyzed respectively in 23 eyes of 18 patients who received LASIK retreatment. The endothelium checked by contact specular microscopy was analyzed for central cell density (CCD), coefficient of cell variation in size and hexagonality preoperatively and 1, 4, 12 weeks after the operation. The amplitude and latency of P(100) in standard pattern visual evoked potentials (P-VEPs), that of a, b waves of different reactions in flash electroretinograms (F-ERGs) and also the total amplitude of oscillatory potentials (OPs) were evaluated preoperatively and 2, 12 weeks after the operation. RESULTS: The mean CCD decreased significantly (3.94%) one week after the operation (P < 0.01). The change was negatively correlated with the thickness of residual untreated posterior cornea (r = -0.719, P < 0.01) and not significantly correlated with the amount of laser pulse (r = 0.371, P > 0.05). There was no significant difference between preoperative and either 4 or 12 week postoperative CCD (P > 0.05). No significant differences in mean coefficient variation and hexagonality were found between preoperative and postoperative values in the follow up (P > 0.05). Examination of electrophysiological function showed that the amplitude of P(100) increased and the latency of P(100) decreased 2 and 12 weeks postoperatively. The significant changes were only at spatial frequency 34' in the P-VEPs test (P < 0.05). In F-ERGs test, there was no significant change in amplitude, latency of a, b waves or total amplitude of OPs between pre-operative and either 2 or 12 week post-operative values (P > 0.05). CONCLUSIONS: Retreatment after LASIK may cause a reversible reduction of CCD with no morphological changes in a short period (1 week) after the operation. It was not absolutely safe for corneal endothelium when the thickness of residual untreated basal cornea approaches to 200 micro m. These results show that the operation seems not to cause functional impairment in the retina and optic nerve. And it also can improve the quality of vision for some degrees. PMID- 12654224 TI - [Clinical manifestations and diagnosis of Vogt-Koyanagi-Harada syndrome]. AB - OBJECTIVE: To investigate the clinical features, causes of blindness and diagnosis of Vogt-Koyanagi-Harada (VKH) syndrome. METHODS: The data of 157 patients with VKH syndrome were reviewed and analyzed. Patients were carefully examined with slit lamp, ophthalmoscope, three-mirror lens, fundus fluorescein angiography (FFA), indocyanine green angiography (ICGA) and HLA typing. RESULTS: Headache was noted in 73.5% of these patients. Simultaneous involvement of both eyes occurred in 80.8% of these patients. Choroiditis, papilledema and edema of the retina adjacent to the optic nerve were noted in 100% of these patients in the posterior uveitis stage, whereas recurrent granulomatous anterior uveitis (98.4%), "sunset glow" fundus (95.8%) and Dalen-Fuchs nodules (71.2%) were the common ocular findings in the recurrent anterior uveitis stage. The common causes of blindness were papillitis, exudative retinal detachment and complicated cataract in the posterior uveitis stage, anterior uveal involvement stage and its recurrent stage. Poliosis (36.3%) and alopecia (35.0%) were the most common extraocular findings. Early irregular patches of fluorescence, followed by localized hyperfluorescent spots were the typical findings of FFA. Dilation of choroidal vessels and leakage of ICG from the choroidal vessels were the common ICGA findings. The prevalence of HLA-DR4 and HLA-DRw53 in patients (54.9% and 71.8% respectively) was significantly higher than that in controls (14.7% and 38.2% respectively). CONCLUSIONS: VKH syndrome is characterized by choroiditis, papillitis or neuroretinitis in the posterior uveitis stage, followed by a generalized uveitis with a typical recurrent granulomatous anterior uveitis. Extraocular findings and relevant examinations including FFA, ICGA and HLA typing are helpful to the diagnosis of VKH syndrome. PMID- 12654225 TI - [Corneal topography and thickness in keratoconus]. AB - OBJECTIVE: To evaluate the corneal topography and pachymetry in keratoconus. METHODS: Thirty-five patients (47 eyes) with keratoconus were evaluated by Orbscan corneal topography system. The location, anterior and posterior surface elevation, curvature and pachymetry were analyzed with special reference to the central point of cornea, the apex, the highest point of posterior surface, the point with greatest curvature and the thinnest point. The correlation between the point of greatest curvature and the apex was also analyzed. RESULTS: The mean elevation of the apex was (0.102 +/- 0.044) mm, the mean distance between the apex and the central point of cornea was (0.686 +/- 0.376) mm. Most of the apexes, the points with the highest posterior surface, the points with the greatest curvature were located in the inferior temporal quadrant, followed by inferior nasal, superior temporal and superior nasal quadrant. The SimK of keratocunus was (60.9 +/- 10.4/53.4 +/- 8.7) D and the mean powers within 3 mm and 5 mm were (57.1 +/- 8.0) D and (50.7 +/- 5.4) D respectively. High correlation between apex elevation and the apex curvature was found (r = 0.697, P < 0.01), and there was also negative relationship between apex elevation and corneal thickness (r = -0.563, P < 0.01). CONCLUSIONS: The Orbscan system can provide accurate and subtle information about the shape and thickness of keratoconus, which can help us to learn this disorder deeply, especially in the early diagnosis of this disease. PMID- 12654226 TI - [An epidemiological survey of low vision and blindness of senile persons in Beixinjing blocks, Shanghai]. AB - OBJECTIVE: To investigate the morbidities of low vision and blindness and the leading cause of blindness in residents aged >or= 60 years in Beixinjing blocks, Shanghai. METHODS: The vision archives were first established for total old residents. The pinhole visual acuity was recorded and classified under WHO grading system. Assigned ophthalmologic doctors assured the leading blind causes of every blind person. RESULTS: The response rates in 1994, 1997 and 2000 were 83.87%, 88.32% and 91.98% respectively. The morbidities of low vision and blindness in these 3 years were 3.61% and 1.51%, 2.31% and 1.09%, 1.31% and 0.71% respectively. The leading causes of blindness were degenerative retinopathy (including age-related maculopathy), cataract, corneal diseases and retinal angiopathy (including hypertensive and diabetic retinopathy, retinal artery or venous occlusion, etc). CONCLUSIONS: The morbidities of low vision and blindness in this area decreased from 1994. The leading cause of blindness was degenerative retinopathy. PMID- 12654227 TI - [An investigation on causes of blindness of children in seven blind schools in East China]. AB - OBJECTIVE: To determine the anatomical sites and underlying causes of blindness and severe visual impairment (BL/SVI), potentially preventable and treatable causes in East China. METHODS: Between April 1998 and May 1999, a total of 385 children attending seven blind school in East China area were examined, and WHO Prevention of Blindness Program (WHO/PBL) Eye Examination Record for children with blindness and low vision was used. RESULTS: Of the 385 children, 356 (92.5%) were blind or visually impaired severely (visual acuity less than 0.1 in the better eye). The commonest anatomical sites of BL/SVI were lens (27.5%), retina (22.5%), whole globe (15.2%) and optic nerve (14.9%). The commonest eye diseases were cataract (26.4%), retinal dystrophy (10.7%), optic nerve hypoplasia (8.4%). The aetiologic causes were hereditary factors (35.1%), meningitis (5.3%), trauma (2.8%), vitamin A deficiency (0.6%) and so on, drugs or alcohol taken during pregnancy (6.2%), retinopathy of premature infant (2.8%). Corneal scarring due to vitamin A deficiency or infection was not a major cause of visual loss. It was considered that 47.5% of cases were avoidable blindness, i.e. 23.0% potentially preventable and 24.4% potentially treatable. CONCLUSIONS: Along with the improvement of health and socio-economic status, nutritional and infective causes of blindness are uncommon, and now hereditary factors become the major causes in children. We shall propagandize rudimentary knowledge of ophthalmology actively, and strengthen the building of pediatric ophthalmology, so as to prevent and treat childhood blindness better. PMID- 12654228 TI - [A new model of choroidal neovascularization in mice]. AB - OBJECTIVE: To investigate the feasibility of inducing choroidal neovascularization (CNV) in mice by diode laser. METHODS: Two hours, 3 days, 1 week, 2 weeks and 4 weeks after photocoagulation, the effect of diode laser for inducing CNV by fundus fluorescein angiography (FFA), light microscopy and transmission electron microscopy. RESULTS: Two hours and 3 days after laser treatment, there were no CNV and fluorescent leakage observed in mice. The first signs of leakage in FFA associated with CNV appeared 1 week after the postphotocoagulation, (48/90 lesions, or 53.3%). Four weeks after laser treatment, there were still 33 lesions appearing leakage (36.67%). Histological sections proved the existence of CNV, but the rate of CNV was not in accord with that of fluorescence leakage. CONCLUSIONS: This animal model is very useful for in vivo studies of angiogenesis and its modulation via various therapy. PMID- 12654229 TI - [The profiles of free organophosphorus poisons in the bile of rabbits poisoned with different organophosphates]. AB - OBJECTIVE: To study the process and significance of the distribution of free organophosphorus poisons (FOPs) in the blood and bile of rabbits poisoned with organophosphates. METHODS: Seventy two livid blue rabbits, male, 2 - 2.5 kg in weight, were divided into 3 groups: trichlorfon (556.0 mg/kg), monocrotophos (11.12 mg/kg) and methyl parathion (37.05 mg/kg). Each group consisted of 24 rabbits. All organophosphates were administered by subcutaneous route. Blood and bile were collected at time intervals of 1, 6, 24, 96 hours after administration. Blood cells and plasma were separated. Acetylcholinesterase (AChE) activity was measured with dithiobisnitrobenzoic acid (DTNB) enzyme kinetic method. The levels of FOPs in plasma and bile were determined with enzyme inhibited method. RESULTS: From 1 h to 96 h after administration negative correlation was found between time and FOP concentration in plasma (trichlorfon, r = -0.74, P < 0.01; monocrotophos, r = -0.55, P < 0.01; methyl parathion, r = -0.69, P < 0.01), and it was also found in bile between time and FOP concentration of trichlorfon (r = -0.97, P < 0.01) and monocrotophos (r = -0.71, P < 0.01). There is no linear correlation between time and concentration of methyl parathion in bile (r = -0.14, P > 0.05). When FOPs in plasma were not detectable at 96 h after administration, high levels of FOPs could still be detected in bile [trichlorfon (300.3 +/- 174.44) IU/L; monocrotophos (362.8 +/- 136.62) IU/L; methyl parathion (101.0 +/- 75.85) IU/L]. CONCLUSION: The bile is the most important store for FOPs in animal. FOPs can exist in bile over 96 h. The process of poisoning is a changing process of absorption, distribution and redistribution and it is different owing to various physical and chemical properties of organophosphates. PMID- 12654230 TI - [The expression and functional characteristics of AC133 antigen in cord blood hematopoietic cells]. AB - OBJECTIVE: To explore the significance of AC(133) antigen expression on cord blood (CB) hematopoietic cells. METHODS: The immunophenotype of CB hematopoietic cells was analyzed by flow cytometric two-color direct immunofluorescence method. Hematopoietic activity of AC(133)(+), CD(34)(+) and AC(133)(-) CD(34)(+) cells in CB was examined by analyzing clonogenic capacity, number of LTC-IC and ex vivo expansion ability. RESULTS: AC(133)(+) cells consisted of 0.67% of the total CB MNCs. Of the AC(133)(+) cells, 93.4% expressed CD(34) antigen and 94.2% was PKH(26)(+). Although the total number of CFC was the same in the AC(133)(-) CD(34)(+) fraction as in the CD(34)(+) and AC(133)(-) fractions, the number of LTC-IC was much higher in the AC(133)(+) fraction. In addition, the features of the colonies grown from these three fractions were quite different. Approximately 50% of the colonies derived from the AC(133)(+) fraction were CFU-GM, whereas more than 60% of the colonies derived from the CD(34)(+) fraction and 80% of the colonies derived from the AC(133)(-) CD(34)(+) fraction were BFU-E. After 7 d ex vivo expansion, the total number of cells and CFC increased 28.2-fold and 7.1 fold respectively in AC(133)(+) cell populations, 6.8% of the cells was PKH(26)(+). CONCLUSIONS: These findings suggest that the early stem/progenitor cells of CB are enriched in AC(133)(+) fraction, AC(133)(+) cells contain higher proliferative efficiency during ex vivo culture. It can generate either committed progenitors or early stem/progenitors. AC(133) antigen would be a useful marker for clinical selection of hematopoietic stem/progenitors. PMID- 12654231 TI - [The application of dual fusion interphase fluorescence in situ hybridization probe in detecting minimal residual disease in chronic myeloid leukemia]. AB - OBJECTIVE: To explore the sensitivity, specificity and reproducibility of dual color and dual-fusion fluorescence in situ hybridization (D-FISH) in detecting the minimal residual disease (MRD) of chronic myeloid leukemia (CML) after treatment. METHODS: MRD were detected in the bone marrow of 8 cases of chronic myeloid leukemia after treatment with interphase fluorescence in situ hybridization. Sensitivity, specificity and reproducibility of D-FISH were compared with those of single-fusion FISH (S-FISH). Furthermore, correlation between the results of D-FISH and RT-PCR was explored. RESULTS: D-FISH was more sensitive and specific than S-FISH and had a good reproducibility. In normal control, the cutoff rates of D-FISH and S-FISH were 0.570% and 5.868% with significant statistic difference. In detection of MRD in 8 cases of CML post transplantation, positive rates of S-FISH and D-FISH were 3/8 (37.5%) and 5/8 (62.5%) respectively. Furthermore, the results of D-FISH had a high correlation with that of RT-PCR. CONCLUSION: In view of the lower false-positive and false negative results of D-FISH, it may be used in the detection of minimal residual disease in CML after treatment. But further exploration is required for verifying the correlation between the results of D-FISH and the clinical parameters. PMID- 12654232 TI - [Association of the cocaine and amphetamine-regulated transcript gene with type 2 diabetes mellitus]. AB - OBJECTIVE: To investigate whether genetic variation in cocaine and amphetamine regulated transcript (CART) gene might contribute to the genesis of type 2 diabetes. METHODS: Screening for mutations in the entire coding region for the CART gene were performed with polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) in 180 normoglycemic control subjects and 221 patients with type 2 diabetes. RESULTS: (1) Adenine deletion was identified at position 1,457 nucleotide located at untranslation area of exon 3. In normal weight control, the frequencies of CART-A + and CART-A-alleles were 83.6% and 16.4% respectively. The frequencies of CART-A + A +, A + A-, A-A- genotype were 68.9%, 29.4% and 1.7% respectively. (2) In diabetic patients, the frequencies of CART-A + and A-alleles were 84.6% and 15.4% respectively; the frequencies of CART A + A +, A + A-, A-A- genotype were 71.9%, 25.3% and 2.7% respectively. The frequency of A deletion of the CART gene in diabetic patients did not differ significantly from normoglycemic control subjects. (3) Diabetic patients with the A deletion had increased total cholesterol, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol. CONCLUSION: Polymorphism was found in the 3'-untranslated region (Delta A1457) of CART in Chinese. A deletion in CART is not associated with type 2 diabetes, but may contribute to dyslipidemia. PMID- 12654233 TI - [A linkage study of cytotoxic T lymphocyte-associated antigen 4 gene and Graves' disease in northern Chinese Han ethnic]. AB - OBJECTIVE: To determine if the cytotoxic T lymphocyte-associated antigen 4 (CTLA 4) gene, which is located on chromosome 2q31-q33, is the major susceptibility gene for Graves' disease (GD) of northern Chinese Han nationality. METHODS: Five highly polymorphic microsatellite markers spanning the entire region of chromosome 2q31-q33 were employed to screen 54 families with multiple incidences of GD (322 individuals) of northern Chinese Han nationality. Tow-point and multi point Lod scores were calculated under different levels of penetrance, assuming both dominant and recessive models. Multipoint nonparametric linkage (NPL) scores were also calculated. RESULTS: Two-point Lod scores (theta = 0) and multipoint Lod scores of less than -2 were observed for all the markers tested, at all levels of penetrance, in both the dominant and recessive models of inheritance. P values of greater than 0.05 were observed for all the multipoint NPL scores obtained. CONCLUSION: We obtained evidence against the linkage of GD with chromosome 2q31-q33, thus we are of the opinion that CTLA-4 gene is not the major susceptibility locus for Graves' disease of northern Chinese Han nationality. PMID- 12654234 TI - [A control study on bacterial resistance and clinical features of lower respiratory tract infection with Pseudomonas aeruginosa in medical intensive care unit and general ward]. AB - OBJECTIVE: To compare the bacterial resistance and clinical features of lower respiratory tact infection with Pseudomonas aeruginosa (PA) in medical intensive care units (MICU) and general wards (GW) and to analyze the predisposing factors to PA infection. METHODS: Three hundred and twenty three patients with lower respiratory tract PA infection, aged from 30 to 91 years, admitted into Shanghai Zhongshan hospital from May 1997 to December 2001 (163 in MICU and 160 in GW) were included in this retrospective study. The predisposing factors, underlying diseases, clinical presentations, laboratory data, treatment and outcome of these patients were analyzed. On the other hand, the susceptibility patterns to antibiotics of 1 387 PA strains (879 from MICU and 508 from GW) from the recorders of the bacteria culture chamber were investigated. RESULTS: The underlying diseases of the patients mainly were chronic obstructive pulmonary disease (23.3% of patients in MICU and 12.5% in GW) and malignant tumor (14.7% in MICU and 14.4% in GW). A relatively high resistant rates of PA strains to common antibiotics were found in both groups: cefoperazone/sulbactam 1.0% (MICU: 1.0%, GW: 1.1%), piperacillin/tazobactam 2.5% (MICU: 1.0%, GW: 5.0%), amikacin 6.1% (MICU: 7.0%, GW: 4.5%), ceftazidime 9.2% (MICU: 11.6%, GW: 2.7%), cefepime 11.3% (MICU: 12.2%, GW: 10.8%), imipenem 13.5% (MICU: 6.8%, GW: 25.2%), piperacillin 13.8% (MICU: 17.8%, GW: 6.9%), cefoperazone 16.4% (MICU: 22.1%, GW: 6.6%), ticarcillin/clavulanic acid 17.2% (MICU: 21.2%, GW: 17.3%), cidomycin 18.5% (MICU: 20.5%, GW: 14.9%), ciprofloxacin 20.2% (MICU: 27.2%, GW: 8.1%), ceftriaxone 44.4% (MICU: 53.6%, GW: 28.4%). Most of the resistant rates of strains from MICU were higher than those from GW. The susceptibilities of PA to ceftazidime and piperacillin changed little during the past 5 years and were above 80.00%. Since 1988, especially from 2000, the susceptibility of PA to most antibiotics became remarkably lower. With selected antibiotic treatment of the infections 196 patients were cured, 79 patients died in hospital. The mortality of patients in GW (16.25%, 26/160) was lower than that in MICU (32.52%, 53/163). CONCLUSION: Precautions to PA infection is important in patients with predisposing factors. Patients with Lower respiratory tract PA infection in MICU had different clinical features, laboratory data and outcome compared to GW patients. The PA isolated from MICU patients had a higher resistant rate to many antibiotics than that from GW patients. It is important to perform antibiotic sensitivity test timely to help correct selection of drugs, especially for MICU patients. PMID- 12654235 TI - [Tumor necrosis factor alpha HLA-DRB(1) gene polymorphism and genetic susceptibility to cirrhosis]. AB - OBJECTIVE: To study the relationship between the gene polymorphism of HLA-DRB(1) and tumor necrosis factor (TNF)alpha with the genetic susceptibility to cirrhosis. METHODS: The gene polymorphism of DRB(1) and TNF alpha of 106 cases of cirrhosis due to HBV and 108 controls were detected by means of polymerase chain reaction-sequence specific primer and RFLP techniques. RESULTS: The frequency of DRB(1) * 120X and TNF2/1 was increased in the patients as compared with the controls (35.9% vs 11.1%, P < 0.001, 19.8% vs 10.2%, P < 0.05 respectively), The frequency of DRB(1) * 150X allele was reduced in the patients as compared with the controls (13.2% vs 30.6%, P < 0.05). It is suggested that the gene polymorphism of HLA-DRB(1) and TNF alpha to be associated with genetic susceptibility to cirrhosis. Cross analysis showed that DRB(1) * 120X allele was more strongly associated with genetic susceptibility to cirrhosis than TNF2 allele. CONCLUSIONS: The genetic susceptibility to cirrhosis is associated with DRB(1) * 120X and TNF2 allele, persons with DRB(1) * 120X and TNF2 allele have an increased risk for the liver cirrhosis occurrence; DRB(1) * 120X allele may be a susceptibility gene to liver cirrhosis and DRB(1) * 150X allele may be a protective gene from liver cirrhosis. PMID- 12654236 TI - [A clinical study of reflux oesophagites]. AB - OBJECTIVE: To study the incidence and role of acid and bile reflux in gastroesophageal reflux disease (GERD). METHODS: Fifty two patients with reflux symptoms were studied. All of them were performed endoscopy received acid and bile reflux monitoring. RESULTS: Fifty two patients received acid and bile reflux monitoring, 42 patients had reflux with the main type of double-reflux (both acid and bile reflux) which was 59.5% (25/42), and with the type of the single-reflux which was 40.5% (17/42). Among single-reflux, the acid reflux was the main type about 76.4% (13/17); Among double-reflux, the degree of acid reflux was more severe, than did bile reflux; The percent of reflux oesophagites join the double reflux patients was significant high than that in single-acid-reflux patients (P < 0.05); There was a liner correlation between acid reflux index either the frequency or the severity of reflux symptoms. CONCLUSIONS: Double-reflux was the main type of reflux in GERD, and acid reflux was the main type among double reflux and single-reflux. There may be a synergistic effect between acid and bile reflux. Acid reflux was the main factor for reflux symptoms. PMID- 12654237 TI - [The renal protective effect of selective cyclooxygenase-2 inhibitor on obstructive nephropathy]. AB - OBJECTIVE: To observe the effect of selective cyclooxygenase-2 (COX-2) inhibitor meloxicam on the progression of tubulointerstitial fibrosis in rats with unilateral ureteral obstruction (UUO). METHODS: UUO rats were treated with meloxicam (M), indomethacin (I) or vehicle alone (U) for 4 weeks. Using reverse transcription-polymerase chain reaction (RT-PCR), we examined the mRNA expressions of transforming growth factor (TGF)-beta receptor-1 and 2 in left kidney from rats with UUO. Immuno-precipitation and immunohistochemistry analysis were carried out to investigate the protein level of TGF-beta(1). Tubulointerstitial fibrosis was quantified by Masson's staining. Similar studies were performed in another group of rats with sham operation (C). RESULTS: mRNA expressions of TGF-beta receptor-1 and 2 were markedly increased in Group U compared with those in Group C. Both meloxicam and indomethacin inhibited their expressions to a different degree (P < 0.01), but the effect of meloxicam was more significant. Meloxicam decreased the protein level of TGF-beta(1) (P < 0.01), while indomethacin had no such effect. Compared with Group C, there appeared tubulointerstitial fibrosis in the kidney of rats with UUO. The lesion was aggravated by indomethacin, but remarkably attenuated by meloxicam. CONCLUSIONS: In summary, selective COX-2 inhibitor-meloxicam has a salutary effect on the tubular and interstitial response to UUO. TGF-beta and its receptor approaches partly explain some of the mechanism. PMID- 12654238 TI - [Relationship between the heterotopic neurons and intractable epilepsy]. AB - OBJECTIVE: To discuss the relationship between heterotopic neurons and intractable epilepsy. METHODS: Epileptogenic zones were precisely located with MRI, video-EEG monitoring, intracranial electrical recording and electrocorticography monitoring and then resected in seven patients. The removed tissue was processed for conventional pathological examination with immunohistochemical staining. RESULTS: Heterotopic neurons were found in the white matter of the specimens. All the seven cases experienced a favorable outcome, with six seizure free. CONCLUSION: The heterotopic neurons scattered or clustered in the white matter contribute to the pathogenesis of intractable epilepsy. PMID- 12654239 TI - Treatment of established osteoporosis: a systematic review and cost-utility analysis. PMID- 12654240 TI - Variation on an Src-like theme. AB - The modularity of protein architecture and the diversity of protein domains hint at a vast combinatorial richness. But evolution appears to have been relatively conservative about selecting new combinations. When a particular grouping of domains within a polypeptide chain can perform a concerted function, that combination tends to reappear in multiple genomic contexts. In other words, once a molecular solution to a functional problem has emerged, it is reused rather than reinvented. PMID- 12654241 TI - Interchangeable parts of the Escherichia coli recombination machinery. AB - In Escherichia coli, at least two groups of proteins, or "recombination machines," can operate independently on broken DNA to produce a 3'-terminated single-stranded DNA filament coated with RecA protein and ready for synapsis with intact homologous DNA. Recent analyses of mutants lacking one or more of the activities required for presynaptic filament formation by one recombination machine demonstrate that parts of the two normally separate machines can interchange to initiate homologous recombination. PMID- 12654242 TI - Membranes, trafficking, and signaling during animal development. AB - Molecular genetics has been key in allowing developmental biologists to uncover many of the molecules that participate in pattern formation. Cell biology is now beginning to help developmental biologists in their quest to understand how these molecules interact within cells to direct tissue behavior. This is particularly true in the areas of membrane trafficking and cell motility. Recent work has shown that various trafficking events such as secretion, endocytosis, segregation in membrane microdomains, intracellular transport, and targeting to lysosomes regulate various signaling pathways. It is likely that within the context of an embryo, these trafficking events are integrated such that secreted factors reliably orchestrate many developmental decisions. PMID- 12654243 TI - Global chromosome positions are transmitted through mitosis in mammalian cells. AB - We investigated positioning of chromosomes during the cell cycle in live mammalian cells with a combined experimental and computational approach. By non invasive labeling of chromosome subsets and tracking by 4D imaging, we could show that no global rearrangements occurred in interphase. Using the same assay, we also observed a striking order of chromosomes throughout mitosis. By contrast, our computer simulation based on stochastic movements of individual chromosomes predicted randomization of chromosome order in mitosis. In vivo, a quantitative assay for single chromosome positioning during mitosis revealed strong similarities between daughter and mother cells. These results demonstrate that global chromosome positions are heritable through the cell cycle in mammalian cells. Based on tracking of labeled chromosomes and centromeres during chromosome segregation and experimental perturbations of chromosomal order, we propose that chromosome specific timing of sister chromatid separation transmits chromosomal positions from one cell generation to the next. PMID- 12654244 TI - Chromosomal cohesin forms a ring. AB - The cohesin complex is essential for sister chromatid cohesion during mitosis. Its Smc1 and Smc3 subunits are rod-shaped molecules with globular ABC-like ATPases at one end and dimerization domains at the other connected by long coiled coils. Smc1 and Smc3 associate to form V-shaped heterodimers. Their ATPase heads are thought to be bridged by a third subunit, Scc1, creating a huge triangular ring that could trap sister DNA molecules. We address here whether cohesin forms such rings in vivo. Proteolytic cleavage of Scc1 by separase at the onset of anaphase triggers its dissociation from chromosomes. We show that N- and C terminal Scc1 cleavage fragments remain connected due to their association with different heads of a single Smc1/Smc3 heterodimer. Cleavage of the Smc3 coiled coil is sufficient to trigger cohesin release from chromosomes and loss of sister cohesion, consistent with a topological association with chromatin. PMID- 12654245 TI - Pirh2, a p53-induced ubiquitin-protein ligase, promotes p53 degradation. AB - The p53 tumor suppressor exerts anti-proliferative effects in response to various types of stress including DNA damage and abnormal proliferative signals. Tight regulation of p53 is essential for maintaining normal cell growth and this occurs primarily through posttranslational modifications of p53. Here, we describe Pirh2, a gene regulated by p53 that encodes a RING-H2 domain-containing protein with intrinsic ubiquitin-protein ligase activity. Pirh2 physically interacts with p53 and promotes ubiquitination of p53 independently of Mdm2. Expression of Pirh2 decreases the level of p53 protein and abrogation of endogenous Pirh2 expression increases the level of p53. Furthermore, Pirh2 represses p53 functions including p53-dependent transactivation and growth inhibition. We propose that Pirh2 is involved in the negative regulation of p53 function through physical interaction and ubiquitin-mediated proteolysis. Hence, Pirh2, like Mdm2, participates in an autoregulatory feedback loop that controls p53 function. PMID- 12654246 TI - Structural basis for the function of the beta subunit of the eukaryotic signal recognition particle receptor. AB - Protein translocation across and insertion into membranes is a process essential to all life forms. In higher eukaryotes, this process is initiated by targeting the translating ribosome to the endoplasmic reticulum via the signal recognition particle (SRP) and its membrane-associated heterodimeric receptor (SR). This targeting step is regulated by three G proteins, SRP54, SR alpha, and SR beta, which act in concert. Little is known about the regulatory role of SR beta. Here, we present the 1.7 A crystal structure of the SR beta-GTP subunit in complex with the interaction domain of SR alpha. Strikingly, the binding interface overlaps largely with the switch 1 region of SR beta. This finding, together with additional biochemical data, shows that the eukaryotic SR is a conditional and not an obligate heterodimer. The results suggest that the GTP/GDP switch cycle of SR beta functions as a regulatory switch for the receptor dimerization. We discuss the implications for the translocation pathway. PMID- 12654247 TI - Different levels of the homeodomain protein cut regulate distinct dendrite branching patterns of Drosophila multidendritic neurons. AB - Functionally similar neurons can share common dendrite morphology, but how different neurons are directed into similar forms is not understood. Here, we show in embryonic and larval development that the level of Cut immunoreactivity in individual dendritic arborization (da) sensory neurons correlates with distinct patterns of terminal dendrites: high Cut in neurons with extensive unbranched terminal protrusions (dendritic spikes), medium levels in neurons with expansive and complex arbors, and low or nondetectable Cut in neurons with simple dendrites. Loss of Cut reduced dendrite growth and class-specific terminal branching, whereas overexpression of Cut or a mammalian homolog in lower-level neurons resulted in transformations toward the branch morphology of high-Cut neurons. Thus, different levels of a homeoprotein can regulate distinct patterns of dendrite branching. PMID- 12654248 TI - ANKTM1, a TRP-like channel expressed in nociceptive neurons, is activated by cold temperatures. AB - Mammals detect temperature with specialized neurons in the peripheral nervous system. Four TRPV-class channels have been implicated in sensing heat, and one TRPM-class channel in sensing cold. The combined range of temperatures that activate these channels covers a majority of the relevant physiological spectrum sensed by most mammals, with a significant gap in the noxious cold range. Here, we describe the characterization of ANKTM1, a cold-activated channel with a lower activation temperature compared to the cold and menthol receptor, TRPM8. ANKTM1 is a distant family member of TRP channels with very little amino acid similarity to TRPM8. It is found in a subset of nociceptive sensory neurons where it is coexpressed with TRPV1/VR1 (the capsaicin/heat receptor) but not TRPM8. Consistent with the expression of ANKTM1, we identify noxious cold-sensitive sensory neurons that also respond to capsaicin but not to menthol. PMID- 12654249 TI - Mechanisms of autoinhibition and STI-571/imatinib resistance revealed by mutagenesis of BCR-ABL. AB - The Bcr-Abl fusion protein kinase causes chronic myeloid leukemia and is targeted by the signal transduction inhibitor STI-571/Gleevec/imatinib (STI-571). Sequencing of the BCR-ABL gene in patients who have relapsed after STI-571 chemotherapy has revealed a limited set of kinase domain mutations that mediate drug resistance. To obtain a more comprehensive survey of the amino acid substitutions that confer STI-571 resistance, we performed an in vitro screen of randomly mutagenized BCR-ABL and recovered all of the major mutations previously identified in patients and numerous others that illuminate novel mechanisms of acquired drug resistance. Structural modeling implies that a novel class of variants acts allosterically to destabilize the autoinhibited conformation of the ABL kinase to which STI-571 preferentially binds. This screening strategy is a paradigm applicable to a growing list of target-directed anti-cancer agents and provides a means of anticipating the drug-resistant amino acid substitutions that are likely to be clinically problematic. PMID- 12654250 TI - A myristoyl/phosphotyrosine switch regulates c-Abl. AB - The c-Abl tyrosine kinase is inhibited by mechanisms that are poorly understood. Disruption of these mechanisms in the Bcr-Abl oncoprotein leads to several forms of human leukemia. We found that like Src kinases, c-Abl 1b is activated by phosphotyrosine ligands. Ligand-activated c-Abl is particularly sensitive to the anti-cancer drug STI-571/Gleevec/imatinib (STI-571). The SH2 domain phosphorylated tail interaction in Src kinases is functionally replaced in c-Abl by an intramolecular engagement of the N-terminal myristoyl modification with the kinase domain. Functional studies coupled with structural analysis define a myristoyl/phosphotyrosine switch in c-Abl that regulates docking and accessibility of the SH2 domain. This mechanism offers an explanation for the observed cellular activation of c-Abl by tyrosine-phosphorylated proteins, the intracellular mobility of c-Abl, and it provides new insights into the mechanism of action of STI-571. PMID- 12654251 TI - Structural basis for the autoinhibition of c-Abl tyrosine kinase. AB - c-Abl is normally regulated by an autoinhibitory mechanism, the disruption of which leads to chronic myelogenous leukemia. The details of this mechanism have been elusive because c-Abl lacks a phosphotyrosine residue that triggers the assembly of the autoinhibited form of the closely related Src kinases by internally engaging the SH2 domain. Crystal structures of c-Abl show that the N terminal myristoyl modification of c-Abl 1b binds to the kinase domain and induces conformational changes that allow the SH2 and SH3 domains to dock onto it. Autoinhibited c-Abl forms an assembly that is strikingly similar to that of inactive Src kinases but with specific differences that explain the differential ability of the drug STI-571/Gleevec/imatinib (STI-571) to inhibit the catalytic activity of Abl, but not that of c-Src. PMID- 12654253 TI - Apoptosis, proliferation, differentiation: in search of the order. AB - In search of the order, we are tempted to universally link cell death, proliferation, differentiation, and senescence. Current models (classical, conflicting signal and quantitative signal models) are restricted, precisely because they attempt to hardware a plethora of end-points of cellular responses. By defining each cellular process in molecular term, one can disconnect proliferation (CDK activation), apoptosis (caspase activation), and differentiation (tissue function genes expression), even though these responses are linked by upstream signal transduction pathways. These ambivalent pathways (e.g. mitogen-activated pathways) simultaneously transduce opposite signals (for growth arrest and cycling, for cell death and survival), which are ultimately translated in all possible combinations of cellular responses. When depicted in multidimensional axis, this universal model may also include invasiveness, senescence, metastatic and angiogenic responses and even such integral characteristics as malignant transformation. PMID- 12654254 TI - NF-kappaB in cancer: a marked target. AB - The imbalance between proliferation and programmed cell death (apoptosis) is one of the critical cellular events that lead to oncogenesis. While there is no doubt that uncontrolled cell proliferation is essential for the development of cancer, deregulation of apoptosis may play an equally important role in this process. Inhibition of apoptosis prevents the death of tumor cells with DNA damage either associated with carcinogenic initiation or cancer therapy. The transcription factor NF-kappaB is a key regulator in oncogenesis. By promoting proliferation and inhibiting apoptosis, NF-kappaB tips the balance between proliferation and apoptosis toward malignant growth in tumor cells. PMID- 12654255 TI - The role of the Bcl-2 protein family in cancer. AB - Seminal studies on the proto-oncogene bcl-2 have first demonstrated that mutations that inhibit programmed cell death (apoptosis) can promote lymphomagenesis and influence the sensitivity of tumour cells to chemotherapy or radiotherapy. It is now widely believed that neoplastic transformation of many, perhaps even all, cell types requires mutational changes that interfere with the cell death programme. In this review, we describe current knowledge of the molecular control of cell death and discuss the role of pro- and anti-apoptotic members of the Bcl-2 protein family in tumourigenesis and anti-cancer therapy. PMID- 12654256 TI - Defects in the apoptotic machinery of cancer cells: role in drug resistance. AB - The therapeutic goal in cancer treatment is to trigger tumor-selective cell death. Since many antineoplastic agents induce an apoptotic type of death in susceptible cells, it is likely that dysfunction of the apoptotic machinery might be an important determinant of resistance to anticancer drugs. Here we review known differences in the apoptotic machinery in cancer cells, and how this knowledge can be used to increase the efficiency of tumor treatment. PMID- 12654257 TI - Cell surface Death Receptor signaling in normal and cancer cells. AB - The extrinsic cell death pathway is initiated upon ligand-receptor interactions at the cell surface including FAS ligand-FAS/APO1, TNF-TNF receptors, and TRAIL TRAIL receptors. Abnormalities of various components of these pathways have been identified in human cancer including loss of FAS expression, deletion or loss of TRAIL receptor DR4, mutation of TRAIL receptor DR5, overexpression of TRAIL decoy TRID or overexpression of Fas decoy, as well as overexpression of the caspase activation inhibitor, FLIP. Death ligands have been explored as potential therapeutics in cancer therapy with some limitations in the case of FAS and TNF due to toxicities. TRAIL remains promising as a therapeutic and has potential for combination with chemo- or radio-therapy. The death receptor signaling pathways include cross-talk with the mitochondrial pathway and can in some cases be influenced by mitochondrial membrane potential changes or NF-kappaB. FLIP and BCL XL expression may reduce sensitivity of cancer cells to combination therapies. PMID- 12654258 TI - Apoptosis in hematological disorders. AB - Apoptosis, the cell's intrinsic death program, plays a critical role in the regulation of tissue homeostasis, especially in cell systems with a high turnover rate such as hematopoietic cells. Imbalances between proliferation and cell death may result in premature death, uncontrolled polyclonal or monoclonal expansion or tumor formation. Also, cytotoxic therapy and immunotherapy of leukemia and lymphoma predominantly mediates cell death through induction of apoptosis. Understanding the molecular events by which tumor cells evade apoptotic deletion and which induce apoptosis by cytotoxic therapies, have provided a paradigm to link normal growth control, malignant transformation and response to therapy. PMID- 12654259 TI - Angiogenesis and apoptosis. AB - This review assembles the laboratory and clinical evidence that cytotoxic chemotherapy and antiangiogenic therapy are each dependent on endothelial cell apoptosis. During cytotoxic chemotherapy, apoptosis of endothelial cells in the vascular bed of tumors precedes apoptosis of tumor cells, even when the tumor has been made drug resistant. Administration of an angiogenesis inhibitor which is not directly cytotoxic to tumor cells can increase tumor cell apoptosis and inhibit tumor growth by inhibiting endothelial proliferation and migration and/or by inducing endothelial apoptosis. Furthermore, oncogene expression and loss of tumor suppressor gene activity can at once protect tumor cells against apoptosis and increase their angiogenic output. Both of these survival advantages conferred on the tumor can be overcome by antiangiogenic therapy. They can also be overcome by cytotoxic chemotherapy administered on a low dose 'antiangiogenic schedule' which continuously exposes endothelial cells in the tumor bed to the drug. As a result, endothelial apoptosis can be demonstrated to precede tumor cell apoptosis, and tumors regress or are inhibited, whether or not the tumor cells are resistant to the drug, and with little or no host toxicity. In contrast, cytotoxic chemotherapy administered on a 'conventional schedule' of maximal tolerated dose followed by an off-therapy interval, becomes ineffective after drug resistance is acquired. On the basis of these experimental findings, chemotherapy of cancer may possibly be improved-i.e. decreased drug resistance and decreased toxic side-effects-by changing dose and schedule to maximize apoptosis of endothelial cells in the vascular bed of tumors. Further improvement may be achieved by combining angiogenesis inhibitors with 'antiangiogenic chemotherapy'. PMID- 12654260 TI - Restoring apoptosis as a strategy for cancer gene therapy: focus on p53 and mda 7. AB - Understanding the molecular and genetic determinants of cancer will provide unique opportunities for developing rational and effective therapies. Malignant cells are frequently resistant to chemotherapy and radiation induced programmed cell death (apoptosis). This resistance can occur by mutations in the tumor suppressor gene p53. Strategies designed to replace this defective tumor suppressor protein, as well as forced expression of a novel cancer specific apoptosis inducing gene, melanoma differentiation associated gene-7 (mda-7), offer promise for restoring apoptosis in tumor cells. Conditional-replicating viruses that selectively induce cytolysis in tumor cells provides an additional means of targeting cancer cells for destruction. Although these approaches represent works in progress, future refinements will in all likelihood result in the next generation of cancer therapies. PMID- 12654261 TI - Gene silencing by systemic delivery of synthetic siRNAs in adult mice. AB - In mammalian cells, RNA duplexes of 21-23 nucleotides, known as small interfering RNAs (siRNAs) specifically inhibit gene expression in vitro. Here, we show that systemic delivery of siRNAs can inhibited exogenous and endogenous gene expression in adult mice. Cationic liposome-based intravenous injection in mice of plasmid encoding the green fluorescent protein (GFP) with its cognate siRNA, inhibited GFP gene expression in various organs. Furthermore, intraperitoneal injection of anti-TNF-alpha siRNA inhibited lipopolysaccharide-induced TNF-alpha gene expression, whereas secretion of IL1-alpha was not inhibited. Importantly, the development of sepsis in mice following a lethal dose of lipopolysaccharide injection, was significantly inhibited by pre-treatment of the animals with anti TNF-alpha siRNAs. Collectively, these results demonstrate that synthetic siRNAs can function in vivo as pharmaceutical drugs. PMID- 12654262 TI - Structural diversity and isomorphism of hydrogen-bonded base interactions in nucleic acids. AB - The wide structural diversity of RNA results in part from the diversity of non Watson-Crick interactions between bases. To examine the repertoire of possible hydrogen bond interactions among bases, we computed databases of base-pairs and base-triples by systematically matching all possible hydrogen-bond donors and acceptors between bases and evaluating the geometries of each planar configuration. For base-pairs, we find 53 arrangements having at least two hydrogen bonds, including 23 pairs with protonated bases that have not previously been modeled. A comparison with experimentally observed base-pairs reveals an unexpected G:U pair recently observed in the ribosome. For base-triples, we find 840 arrangements in which the three bases are constrained by a total of at least three hydrogen bonds. Base-triples in particular exhibit a wide range of structural diversity, suggesting how compact or elongated nucleic acid structures may be constructed using different hydrogen-bonding patterns. Base-pair and base triple conformations were systematically compared to identify structurally isomorphic combinations, and the experimentally observed arrangements within double and triple helices are among the most isomorphic. Unexpectedly, however, other combinations in the database are even more isomorphic, including several in which all-purine arrangements overlap with all-pyrimidine arrangements. These studies highlight some of the combinatoric and geometric versatility of base interactions and help provide a framework for analyzing and modeling isomorphic interactions and potentially for designing novel nucleic acid structures. PMID- 12654263 TI - Recognition of nucleic acid bases and base-pairs by hydrogen bonding to amino acid side-chains. AB - Sequence-specific protein-nucleic acid recognition is determined, in part, by hydrogen bonding interactions between amino acid side-chains and nucleotide bases. To examine the repertoire of possible interactions, we have calculated geometrically plausible arrangements in which amino acids hydrogen bond to unpaired bases, such as those found in RNA bulges and loops, or to the 53 possible RNA base-pairs. We find 32 possible interactions that involve two or more hydrogen bonds to the six unpaired bases (including protonated A and C), 17 of which have been observed. We find 186 "spanning" interactions to base-pairs in which the amino acid hydrogen bonds to both bases, in principle allowing particular base-pairs to be selectively targeted, and nine of these have been observed. Four calculated interactions span the Watson-Crick pairs and 15 span the G:U wobble pair, including two interesting arrangements with three hydrogen bonds to the Arg guanidinum group that have not yet been observed. The inherent donor-acceptor arrangements of the bases support many possible interactions to Asn (or Gln) and Ser (or Thr or Tyr), few interactions to Asp (or Glu) even though several already have been observed, and interactions to U (or T) only if the base is in an unpaired context, as also observed in several cases. This study highlights how complementary arrangements of donors and acceptors can contribute to base-specific recognition of RNA, predicts interactions not yet observed, and provides tools to analyze proposed contacts or design novel interactions. PMID- 12654264 TI - Extending the understanding of mutagenicity: structural insights into primer extension past a benzo[a]pyrene diol epoxide-DNA adduct. AB - DNA polymerase enzymes employ a number of innate fidelity mechanisms to ensure the faithful replication of the genome. However, when confronted with DNA damage, their fidelity mechanisms can be evaded, resulting in a mutation that may contribute to the carcinogenic process. The environmental carcinogen benzo[a]pyrene is metabolically activated to reactive intermediates, including the tumorigenic (+)-anti-benzo[a]pyrene diol epoxide, which can attack DNA at the exocyclic amino group of guanine to form the major (+)-trans-anti-[BP]-N(2)-dG adduct. Bulky adducts such as (+)-trans-anti-[BP]-N(2)-dG primarily block DNA replication, but are occasionally bypassed and cause mutations if paired with an incorrect base. In vitro standing-start primer-extension assays show that the preferential insertion of A opposite (+)-trans-anti-[BP]-N(2)-dG is independent of the sequence context, but the primer is extended preferentially when dT is positioned opposite the damaged base in a 5'-CG*T-3' sequence context. Regardless of the base positioned opposite (+)-trans-anti-[BP]-N(2)-dG, extension of the primer past the lesion site poses the greatest block to polymerase progression. In order to gain insight into primer-extension of each base opposite (+)-trans anti-[BP]-N(2)-dG, we carried out molecular modeling and 1.25 ns unrestrained molecular dynamics simulations of the adduct in the +1 position of the template within the replicative pol I family T7 DNA polymerase. Each of the four bases was modeled at the 3' terminus of the primer, incorporated opposite the adduct, and the next-to-be replicated base was in the active site with its Watson-Crick partner as the incoming nucleotide. As in our studies of nucleotide incorporation, (+)-trans-anti-[BP]-N(2)-dG was modeled in the syn conformation in the +1 position, with the BP moiety on the open major groove side of the primer template duplex region, leaving critical protein-DNA interactions intact. The present work revealed that the efficiency of primer-extension past this bulky adduct opposite each of the four bases in the 5'-CG*T-3' sequence can be rationalized by the stability of interactions between the polymerase protein, primer-template DNA and incoming nucleotide. However, the relative stabilization of each nucleotide opposite (+)-trans-anti-[BP]-N(2)-dG in the +1 position (T > G > A > or = C) differed from that when the adduct and partner were the nascent base-pair (A > T > or = G > C). In addition, extension past (+)-trans-anti-[BP] N(2)-dG may pose a greater block to a high fidelity DNA polymerase than does nucleotide incorporation opposite the adduct because the presence of the modified base-pair in the +1 position is more disruptive to the polymerase-DNA interactions than it is within the active site itself. The dN:(+)-trans-anti-[BP] N(2)-dG base-pair is strained to shield the bulky aromatic BP moiety from contact with the solvent in the +1 position, causing disruption of protein-DNA interactions that would likely result in decreased extension of the base-pair. These studies reveal in molecular detail the kinds of specific structural interactions that determine the function of a processive DNA polymerase when challenged by a bulky DNA adduct. PMID- 12654265 TI - Monomeric complex of human orphan estrogen related receptor-2 with DNA: a pseudo dimer interface mediates extended half-site recognition. AB - While most nuclear receptors bind DNA as homo or heterodimers, the human estrogen related receptors (hERRs) are members of a subfamily of orphan receptors that bind DNA as monomers. We have determined the solution structure of the DNA binding domain (DBD) of hERR2 bound to its cognate DNA. The structure and base interactions of the core DBD are similar to those of other nuclear receptors. However, high-affinity, sequence-specific DNA binding as a monomer necessitates formation of additional base contacts outside the core DBD. This is accomplished using a modified guanosine-binding "AT-hook" within the C-terminal extension (CTE) flanking the DBD, which makes base-specific minor groove interactions. The structure of the CTE is stabilized both by interactions with the DNA and by packing against a region of the core DBD normally reserved for dimerization. This pseudo-dimer interface provides a basis for the expansion of DNA recognition and suggests a mechanism through which dimerization may have evolved from an ancestral monomeric receptor. PMID- 12654266 TI - NMR structure of Citrobacter freundii AmpD, comparison with bacteriophage T7 lysozyme and homology with PGRP domains. AB - AmpD is a bacterial amidase involved in the recycling of cell-wall fragments in Gram-negative bacteria. Inactivation of AmpD leads to derepression of beta lactamase expression, presenting a major pathway for the acquisition of constitutive antibiotic resistance. Here, we report the NMR structure of AmpD from Citrobacter freundii (PDB accession code 1J3G). A deep substrate-binding pocket explains the observed specificity for low molecular mass substrates. The fold is related to that of bacteriophage T7 lysozyme. Both proteins bind zinc at a conserved site and require zinc for amidase activity, although the enzymatic mechanism seems to differ in detail. The structure-based sequence alignment identifies conserved features that are also conserved in the eukaryotic peptidoglycan recognition protein (PGRP) domains, including the zinc-coordination site in several of them. PGRP domains thus belong to the same fold family and, where zinc-binding residues are conserved, may have amidase activity. This hypothesis is supported by the observation that human serum N-acetylmuramyl-L alanine amidase seems to be identical with a soluble form of human PGRP-L. PMID- 12654267 TI - Structural basis for GroEL-assisted protein folding from the crystal structure of (GroEL-KMgATP)14 at 2.0A resolution. AB - Nucleotide regulates the affinity of the bacterial chaperonin GroEL for protein substrates. GroEL binds protein substrates with high affinity in the absence of ATP and with low affinity in its presence. We report the crystal structure of (GroEL-KMgATP)(14) refined to 2.0 A resolution in which the ATP triphosphate moiety is directly coordinated by both K(+) and Mg(2+). Upon the binding of KMgATP, we observe previously unnoticed domain rotations and a 102 degrees rotation of the apical domain surface helix I. Two major consequences are a large lateral displacement of, and a dramatic reduction of hydrophobicity in, the apical domain surface. These results provide a basis for the nucleotide-dependent regulation of protein substrate binding and suggest a mechanism for GroEL assisted protein folding by forced unfolding. PMID- 12654268 TI - Pressure-dependent changes in the solution structure of hen egg-white lysozyme. AB - The "rules" governing protein structure and stability are still poorly understood. Important clues have come from proteins that operate under extreme conditions, because these clarify the physical constraints on proteins. One obvious extreme is pressure, but so far little is known of the behavior of proteins under pressure, largely for technical reasons. We have therefore developed new methodology for calculating structure change in solution with pressure, using NMR chemical shift changes, and we report the change in structure of lysozyme on going from 30 bar to 2000 bar, this being the first solution structure of a globular protein under pressure. The alpha-helical domain is compressed by approximately 1%, due to tighter packing between helices. The interdomain region is also compressed. By contrast, the beta-sheet domain displays very little overall compression, but undergoes more structural distortion than the alpha-domain. The largest volume changes tend to occur close to hydrated cavities. Because isothermal compressibility is related to volume fluctuation, this suggests that buried water molecules play an important role in conformational fluctuation at normal pressures, and are implicated as the nucleation sites for structural changes leading to pressure denaturation or channel opening. PMID- 12654269 TI - Hammond behavior versus ground state effects in protein folding: evidence for narrow free energy barriers and residual structure in unfolded states. AB - Apparent transition state movement upon mutation or changes in solvent conditions is frequently observed in protein folding and is often interpreted in terms of Hammond behavior. This led to the conclusion that barrier regions in protein folding are broad maxima on the free energy landscape. Here, we use the concept of self-interaction and cross-interaction parameters to test experimental data of 21 well-characterized proteins for Hammond behavior. This allows us to characterize the origin of transition state movements along different reaction coordinates. Only one of the 21 proteins shows a small but coherent transition state movement in agreement with the Hammond postulate. In most proteins the structure of the transition state is insensitive to changes in protein stability. The apparent change in the position of the transition state upon mutation, which is frequently observed in phi-value analysis, is in most cases due to ground state effects caused by structural changes in the unfolded state. This argues for significant residual structure in unfolded polypeptide chains of many proteins. Disruption of these residual interactions by mutation often leads to decreased folding rates, which implies that these interactions are still present in the transition state. The failure to detect Hammond behavior shows that the free energy barriers encountered by a folding polypeptide chain are generally rather narrow and robust maxima for all experimentally explorable reaction coordinates. PMID- 12654270 TI - PKA-dependent binding of mRNA to the mitochondrial AKAP121 protein. AB - Protein kinase A (PKA) anchoring proteins (AKAPs) tether PKA to various subcellular locations. AKAP121, which tethers PKAII to the outer mitochondrial membrane, includes a K homology (KH) RNA-binding motif. Purified AKAP121 KH domain binds the 3' untranslated regions (3'UTRs) of transcripts encoding the Fo f subunit of mitochondrial ATP synthase and manganese superoxide dismutase (MnSOD). Binding requires a structural motif in the 3'UTR and is stimulated by PKA phosphorylation of the domain or a mutation that mimics this phosphorylation. AKAP121 expressed in HeLa cells promotes the translocation of MnSOD mRNA from cytosol to mitochondria and an increase in mitochondrial MnSOD. Both reactions are stimulated by cAMP. Thus, by focusing translation at the mitochondrial membrane, AKAP121 may facilitate import of mitochondrial proteins in response to cAMP stimulation. PMID- 12654271 TI - Community-level access and control in the management of Lake Victoria's fisheries. AB - The fisheries of Lake Victoria have reached a critical stage in their development: catches are declining, and there is increasing evidence of hardship amongst the fishing communities. In an effort to countermand the reduction in the flow of economic benefits the lake's riparian states are collaborating with international agencies to develop a fisheries management strategy that would be based on a partnership between stakeholders. One possible area for such collaboration is via the devolution of certain access controls to fishing communities. This paper describes findings from recent research and argues that notions of access are linked to community perceptions of the resource, control over it and their own individual and collective manoeuvrings designed to retain and improve livelihoods. PMID- 12654273 TI - Relevance of joint action toxicity evaluations in setting realistic environmental safe limits of heavy metals. AB - The evaluation of types of toxicological interactions existing between heavy metals, which are prominent in effluents of some industrial establishments in Lagos State, Nigeria and the Lagos lagoon sediment was carried out against benthic animals, Tympanotonus fuscatus, Clibanarius africanus and Sesarma huzardi of the Lagos lagoon. In order to determine the type of interactions existing between the metals, acute toxicity tests of the metal compounds when acting singly and in joint action studies, by adopting mixture ratios that depict (i) the proportions of the concentrations of the metal ions in the sediment of the Lagos lagoon and (ii) equitoxic mixtures, i.e. based on the 96 hLC(50) values of the metal compounds under single action studies were carried out in laboratory biotests. The joint action evaluations of the test metal mixtures, prepared on the basis of the proportion of the availability of the metals ions in lagoon sediment and equitoxic ratio against the test animals agreed mainly with the model of antagonism (reduction in toxicity) except for test mixtures prepared on the basis of equitoxic ratio and tested against T. fuscatus, where interactions between the test metals was in conformity with the model of synergism, indicating that the toxicity of the constituent metals in the mixture was enhanced. Furthermore, on the basis of classification of synergistic ratio model, the toxic effect of Pb compound which was the least toxic metal compound in the single action toxicity studies was found to be enhanced (synergised) in the presence of other metals when tested jointly. The significance of the results in setting water quality criteria aimed at protecting aquatic biota was discussed. PMID- 12654272 TI - Time activity modelling of domestic exposures to radon. AB - Radon gas occurs naturally in the environment with a variable distribution. In some areas radon concentrates sufficiently within the built environment that it is considered as a public health risk. It is possible, successfully, to reduce radon levels in the built environment, and it has been shown that such remediation programmes can be justified in terms of the costs and benefits accruing. However, the estimated dose received by people in their homes depends on the time spent indoors. The research presented here uses data derived from time activity surveys in Northamptonshire, together with radon data from a representative home, to model potential exposures for different population sub groups. Average home occupancy ranged from 14.8h (probable error 2.5h) for students to 17.7 (3.1) h for adults; schoolchildren spent an average of 14.9 (1.2) h at home. Over a quarter of adults, however, were in the home for 22 h on more. These differences in occupancy patterns lead to substantial differences in radon exposure. In a home with an average hourly ground floor radon concentration of 467 Bqm(-3), modelled hourly average exposures ranged from ca. 250 Bqm(-3) for students and school children, to over 340 Bqm(-3), for women based at home. Modelled exposures show a non-linear association with total time spent at home, suggesting that exposure estimates based on linear models may provide misleading estimates of health risks from radon and the potential benefits of radon remediation. Highest hourly exposures are likely to be experienced by people with highly occupancy, living in single-storey, ground floor accommodation (for example, the elderly the infirm and non-working young mothers). Since these may be least aware of radon risks, and least able to take up remediation measures, they should be specifically targeted for radon monitoring and for assistance in remediation schemes. PMID- 12654274 TI - Environmental effects of agri-environmental schemes in Western Europe. AB - Agri-environmental schemes (AES) have been introduced as part of European Union's (EU) Common Agricultural Policy and are now an important part of this. A methodological approach to analyse the policy effects of AES is outlined, in which we distinguish between performance effects (on agricultural practices) and outcome effects (environmental impact). The performance effects are further approached including measurement of improvement and protection effects based on 12 indicators on changes/maintenance of land use and agricultural management. Data from personal interviews of participating and non-participating farmers in AES measures in nine EU Member States and Switzerland were used to analyse policy effects, including single indicator effects on agricultural practices as well as combined effects at the agreement level. Significant effects were found for mineral N-fertiliser use, stocking density reduction, maintenance of a minimum livestock density and pesticides. For AES agreements regulating grassland management, fertiliser use and pesticides, clear indications of combined improvement and protection effects were found. In addition clear improvement effects of agreements regulating fertiliser and pesticides use on mainly arable lands were revealed. It is concluded that the approach presented including the 12 selected indicators has proven to be operational. PMID- 12654275 TI - Win-win opportunities and environmental regulation: testing of porter hypothesis for Indian manufacturing industries. AB - This paper studies the effect of environmental regulation on the productive efficiency of water polluting industries in India. The panel data of 92 firms belonging to sugar industry of India during the period 1996-99 are used to test the Porter hypothesis of having win-win opportunities for the firms subjected to the regulation. The main empirical result is that the technical efficiency of firms increases with the degree of compliance of firms to the environmental regulation and the water conservation efforts there by supporting the Porter hypothesis. PMID- 12654276 TI - A multi-criteria targeting approach to neutral grassland conservation. AB - Resources for creating and managing rare habitats are limited, and a targeting approach aimed at identifying the most viable sites for habitat conservation is therefore desirable. This study developed a multi-criteria targeting approach to site conservation for two rare grassland types, based on a suite of biotic and abiotic factors managed within a Geographical Information System. A number of biotic and abiotic criteria were assessed to evaluate the biodiversity status of grassland sites. Biotic factors included species diversity, species richness and species rarity; and abiotic factors included patch area, position in the ecological unit and the influence of surrounding land use. Each criterion was given equal weighting and a final biodiversity value for each patch was calculated; the patch with the highest cumulative rank score was deemed the patch with the greatest biodiversity. Each site was then examined in relation to agricultural land under the existing management prescriptions of the Upper Thames Tributaries Environmentally Sensitive Area (UTTESA). Sites identified with high biodiversity potential, but currently not included under management prescriptions, were targeted for future inclusion in the ESA scheme. The targeting approach demonstrated how the national Lowland Meadows habitat action plan creation target of 500 ha could be achieved in the UTTESA. The fact that this target figure was so easily attained within this study area highlighted the possible underestimation of national habitat creation targets. PMID- 12654277 TI - Learning through public involvement in environmental assessment hearings. AB - This research examined the opportunities for critical education available to participants of a federal environmental assessment (EA) that includes hearings. The Sable Gas Panel Review, an assessment of a natural gas project situated in the Maritimes, was undertaken between 1996 and 1997. This study documented what participants learned through their experiences and how that learning was facilitated. Primary data collection emphasized the use of semi-structured interviews to record the experientially based observations of panel participants. Primary data were supported by a review of material submitted by hearing participants for consideration by the panel, and complemented with literature related to EA, environmental education, and transformational learning theories. Evidence was considered in terms of assessment-specific operational definitions, developed from Shor (1993) [Education is politics: Paulo Freire's Critical Pedagogy. In: McLaren, P., Leonard, P. (Eds.), Paulo Freire : A critical Encounter, Routledge, New York, pp. 25-35] ten indicators of critical education. The study revealed that participants of the Sable Gas panel review had an opportunity to engage in critical education. While it is important to acknowledge that the panel did not set out to engage participants in a 'critical educational' experience, each of the 10 operational definitions of critical education used in the study were addressed to varying degrees through activities undertaken by the panel secretariat. Results illustrate the types of educational opportunities, such as a class on how to participate in a quasi-judicial hearing, and the importance of participant-led research that can arise out of the panel review process. The findings of this research contribute to the evolving literature regarding the role of critical education in EA and environmental management. Understanding the existing scope of critical education within a panel review provides the foundation for identifying opportunities for expanding the capacity of the existing EA process to facilitate learning by participants. PMID- 12654278 TI - Developing and applying an index of environmental integrity for the US Mid Atlantic region. AB - Environmental conditions in the Mid-Atlantic region of United States are presently being documented in a series of reports that use 'environmental report cards' to summarize the condition of individual natural resources (e.g. estuaries, streams, forests, and landscapes) over the entire region and within major subregions. An 'index of environmental integrity' (IEI) approach has been developed and is illustrated using the information content of these report cards to evaluate the overall condition throughout the region. The IEI approach is a four-step process: (1) select individual components for the index, (2) calculate subindex values for each of the individual components, (3) aggregate the subindex values into the overall index, and (4) interpretation of index values. The IEI approach was illustrated by applying to the Mid-Atlantic estuaries and wadable streams in the Mid-Atlantic Highlands. Because the IEI approach is new, application should not be made without considering issues such as evaluation of the selection of the indicators, weighting scheme, uncertainties, and appropriate way to interpret the values. PMID- 12654281 TI - Apoptosis pathways: turned on their heads? PMID- 12654279 TI - The current and future management of wild mammals hunted with dogs in England and Wales. AB - There is increasing concern about the use of lethal methods to control wild mammal populations, especially those methods that may have significant impacts on animal welfare. The continued use of dogs to hunt wild mammals in England and Wales, principally foxes (Vulpes vulpes), red deer (Cervus elaphus), brown hares (Lepus europaeus) and mink (Mustela vison), has become a focus for political debate and has been the subject of a recent UK government inquiry. This paper reports the results of a questionnaire study to quantify the use, effectiveness and acceptability of the different methods currently used to manage these four species, and future changes in management following a possible ban on hunting with dogs. There was no straightforward relationship between culling pressure and perceived pest status of the different species from the questionnaire data. For foxes and brown hares, the proportion of land managers (practitioners) carrying out lethal control was higher than that considering these species to be pests. However, the reverse was the case for mink. The most frequently used and effective control methods, which were most acceptable to practitioners and public alike, were various forms of shooting. The general public perceived hunting with dogs as one of the least acceptable means of control for all four species. Practitioners thought that hunting with dogs for red deer and the use of terriers against foxes were among the least acceptable forms of control, but considered hunting with dogs in other situations and for other species to be relatively acceptable. Most practitioners said a ban on hunting with dogs would make no difference to their management of the four species. A ban on hunting with dogs would have minimal impact on populations of foxes, red deer and mink, but it may be of conservation benefit to hares. PMID- 12654282 TI - Enhanced oncolytic potency of replicative adenovirus expressing p53. PMID- 12654283 TI - 3D structure of AcrB: the archetypal multidrug efflux transporter of Escherichia coli likely captures substrates from periplasm. AB - Recent advances in structural biology have extended our understanding of the multiple drug efflux complex, AcrAB-TolC, of Escherichia coli. This tripartite complex and its homologs are the major mechanisms that give most Gram-negative bacteria their characteristic intrinsic resistance to a variety of lipophilic drugs, dyes, and detergents. Most recently, the structure of the transporter AcrB was elucidated at high resolution [Nature 419(2002)587]. It is a particularly significant achievement since integral membrane proteins are notoriously elusive structures for crystallography. The striking features of this trimeric pump, such as the presence of potential substrate-binding sites in the periplasmic domain and the possibility of direct interaction with the end of TolC tunnel, refine our understanding of the mode of action of this tripartite efflux transport complex. PMID- 12654284 TI - The use of cyclin-dependent kinase inhibitors alone or in combination with established cytotoxic drugs in cancer chemotherapy. AB - Cyclin-dependent kinase (CDK) inhibitors are small molecule inhibitors of the kinases required for the orderly progression of cells, both normal and neoplastic, through the cell cycle. Because cell cycle dysregulation is such a common occurrence in neoplasia, the search for agents that might block cell cycle traverse has been the focus of intense interest. These efforts have led to the identification of a broad array of compounds that interfere directly with the function of CDKs. Two of these agents (flavopiridol and UCN-01) have now entered the clinical arena, and others are scheduled to do so in the near future. In preclinical studies, CDK inhibitors have shown the ability not only to block neoplastic cell proliferation, but also to induce, through a variety of mechanisms, programmed cell death. The latter capacity may stem from the diverse effects that CDK inhibitors exert on multiple kinases and apoptotic regulatory molecules. In addition, there is abundant preclinical evidence that CDK inhibitors can potentiate, generally in a dose- and sequence-dependent manner, the anti-tumor effects of many established cytotoxic agents. In clinical studies in humans, flavopiridol and UCN-01 have been shown to be tolerable, although clear evidence of single agent activity or enhancement of the efficacy of established agents has not yet emerged. This may reflect a failure to optimize drug schedules/pharmacokinetics, or to identify the critical molecular targets of these agents. Finally, in recent years, a rationale has emerged for combining CDK inhibitors with other molecularly targeted agents (i.e. differentiation-inducers and signal transduction modulators). Current research has basically two goals: (a). to identify CDK inhibitor concentrations and schedules that inhibit the growth of and induce apoptosis in specific tumor cell types; and (b). to establish a rational basis for combining CDK inhibitors with more conventional cytotoxic agents to enhance antitumor efficacy. This review gives a brief summary of such efforts, with an emphasis on agents and combinations that are in or near clinical development. PMID- 12654285 TI - Tumor p53 status and response to topoisomerase II inhibitors. AB - It is thought that when tumor cells are treated with anticancer drugs, they die through the apoptotic pathway and that cell resistance to cancer chemotherapy is mainly a resistance to apoptosis commitment. p53 is not functional in nearly half of the tumors examined and because of its involvement (directly or through its target genes) in the apoptotic pathway, drug resistance to chemotherapy has been largely attributed to the status of this "tumor suppressor protein". Topoisomerase II (topo II) inhibitors are widely used not only as single agents, but also in the majority of combination treatment protocols for hematologic malignancies and solid tumors. The relationship between p53 and topo II raises many questions about basic regulatory, biochemical, structural and functional characteristics that could be different in cells in different tissues, and most importantly, between different tumor cell types and their normal tissue counterpart. Understanding these relationships may lead to strategies for chemotherapy optimization and further precision targeting of tumor cells in order to avoid drug resistance and thereby chemotherapy failure. PMID- 12654286 TI - Insights on antibiotic resistance of Staphylococcus aureus from its whole genome: genomic island SCC. AB - Staphylococci are ubiquitous colonizers of the skin and mucous membranes and Staphylococcus aureus is the most pathogenic species. The spread of antibiotic resistance among S. aureus strains is a major concern in the treatment of staphylococcal infections. Acquisition of resistance may involve mutation of a bacterial gene on the chromosome or transfer of a resistance gene from other organisms by some form of genetic exchange (conjugation, transduction, or transformation). Completion of whole genome sequences of three methicillin resistant S. aureus (MRSA) strains has provided us a bird's-eye view of the distribution of the mobile genetic elements in the bacterial chromosome that encode antibiotic resistance as well as pathogenicity in S. aureus. PMID- 12654288 TI - Windows into development: historic, current, and future perspectives on transgenic zebrafish. AB - The recent explosion of transgenic zebrafish lines in the literature demonstrates the value of this model system for detailed in vivo analysis of gene regulation and morphogenetic movements. The optical clarity and rapid early development of zebrafish provides the ability to follow these events as they occur in live, developing embryos. This article will review the development of transgenic technology in zebrafish as well as the current and future uses of transgenic zebrafish to explore the dynamic environment of the developing vertebrate embryo. PMID- 12654289 TI - Wnt signaling components in the chicken intestinal tract. AB - Wnt signal transduction has emerged as an increasingly complex pathway due to the numerous ligands, receptors, and modulators identified in multiple developmental systems. Wnt signaling has been implicated in the renewal of the intestinal epithelium within adult animals and the progression of cancer in the colon. The Wnt family, however, has not been explored for function during embryonic gut development. Thus, to dissect the role of Wnt signaling in the developing gastrointestinal tract, it is necessary to first obtain a complete picture of the spatiotemporal expression of the Wnt signaling factors with respect to the different tissue layers of the gut. Here, we offer an in depth in situ gene expression study of Wnt ligands, frizzled receptors, and frizzled related modulators over several days of chicken gut development. These data show some expected locations of Wnt signaling as well as a surprising lack of expression of factors in the hindgut. This paper describes the first comprehensive characterization of the dynamic expression of Wnt signaling molecules during gut development. These data form the basis for future studies to determine the role of Wnt signaling in the developing gastrointestinal tract. PMID- 12654290 TI - BMP receptor 1b is required for axon guidance and cell survival in the developing retina. AB - Previous work has documented the importance of BMPs in eye development. Loss-of function studies in mice, with targeted deletions in either the Bmp7 or Bmp4 genes, have shown that these molecules are critical for early eye development. On the basis of the asymmetry in the dorsal-ventral expression patterns of several members of this family, it has been proposed that these molecules are critical for some aspect of dorsal-ventral patterning in the eye; however, it has been difficult to test this hypothesis because of the early requirement for BMPs in eye development. We have therefore examined the effects of loss of one of the BMP receptors, the BmprIb, on the development of the eye by using targeted deletion. We have found that BmprIb is expressed exclusively in the ventral retina during embryonic development and is required for normal ventral ganglion cell axon targeting to the optic nerve head. In mice with a targeted deletion of the BmprIb gene, many axons arising from the ventrally located ganglion cells fail to enter the optic nerve head, and instead, make abrupt turns in this region. A second phenotype in these mice is a significantly elevated inner retinal apoptosis during a distinct phase of postnatal development, at the end of neurogenesis. Our results therefore show two distinct requirements for BmprIb in mammalian retinal development. PMID- 12654291 TI - Eyeless collaborates with Hedgehog and Decapentaplegic signaling in Drosophila eye induction. AB - eyeless (ey) is a key regulator of the eye development pathway in Drosophila. Ectopic expression of ey can induce the expression of several eye-specification genes (eya, so, and dac) and induce eye formation in multiple locations on the body. However, ey does not induce eye formation everywhere where it is ectopically expressed, suggesting that EY needs to collaborate with additional factors for eye induction. We examined ectopic eye induction by EY in the wing disc and found that eye induction was spatially restricted to the posterior compartment and the anterior-posterior (A/P) compartmental border, suggesting a requirement for both HH and DPP signaling. Although EY in the anterior compartment induced dpp and dac, these were not sufficient for eye induction. Coexpression experiments show that EY needs to collaborate with high level of HH and DPP to induce ectopic eye formation. Ectopic eye formation also requires the activation of an eye-specific enhancer of the endogenous hh gene. PMID- 12654292 TI - T1alpha, a lung type I cell differentiation gene, is required for normal lung cell proliferation and alveolus formation at birth. AB - T1alpha, a differentiation gene of lung alveolar epithelial type I cells, is developmentally regulated and encodes an apical membrane protein of unknown function. Morphological differentiation of type I cells to form the air-blood barrier starts in the last few days of gestation and continues postnatally. Although T1alpha is expressed in the foregut endoderm before the lung buds, T1alpha mRNA and protein levels increase substantially in late fetuses when expression is restricted to alveolar type I cells. We generated T1alpha null mutant mice to study the role of T1alpha in lung development and differentiation and to gain insight into its potential function. Homozygous null mice die at birth of respiratory failure, and their lungs cannot be inflated to normal volumes. Distal lung morphology is altered. In the absence of T1alpha protein, type I cell differentiation is blocked, as indicated by smaller airspaces, many fewer attenuated type I cells, and reduced levels of aquaporin-5 mRNA and protein, a type I cell water channel. Abundant secreted surfactant in the narrowed airspaces, normal levels of surfactant protein mRNAs, and normal patterns and numbers of cells expressing surfactant protein-B suggest that differentiation of type II cells, also alveolar epithelial cells, is normal. Anomalous proliferation of the mesenchyme and epithelium at birth with unchanged numbers of apoptotic cells suggests that loss of T1alpha and/or abnormal morphogenesis of type I cells alter the proliferation rate of distal lung cells, probably by disruption of epithelial-mesenchymal signaling. PMID- 12654293 TI - ePAD, an oocyte and early embryo-abundant peptidylarginine deiminase-like protein that localizes to egg cytoplasmic sheets. AB - Selected for its high relative abundance, a protein spot of MW approximately 75 kDa, pI 5.5 was cored from a Coomassie-stained two-dimensional gel of proteins from 2850 zona-free metaphase II mouse eggs and analyzed by tandem mass spectrometry (TMS), and novel microsequences were identified that indicated a previously uncharacterized egg protein. A 2.4-kb cDNA was then amplified from a mouse ovarian adapter-ligated cDNA library by RACE-PCR, and a unique 2043-bp open reading frame was defined encoding a 681-amino-acid protein. Comparison of the deduced amino acid sequence with the nonredundant database demonstrated that the protein was approximately 40% identical to the calcium-dependent peptidylarginine deiminase (PAD) enzyme family. Northern blotting, RT-PCR, and in situ hybridization analyses indicated that the protein was abundantly expressed in the ovary, weakly expressed in the testis, and absent from other tissues. Based on the homology with PADs and its oocyte-abundant expression pattern, the protein was designated ePAD, for egg and embryo-abundant peptidylarginine deiminase-like protein. Anti-recombinant ePAD monospecific antibodies localized the molecule to the cytoplasm of oocytes in primordial, primary, secondary, and Graafian follicles in ovarian sections, while no other ovarian cell type was stained. ePAD was also expressed in the immature oocyte, mature egg, and through the blastocyst stage of embryonic development, where expression levels began to decrease. Immunoelectron microscopy localized ePAD to egg cytoplasmic sheets, a unique keratin-containing intermediate filament structure found only in mammalian eggs and in early embryos, and known to undergo reorganization at critical stages of development. Previous reports that PAD-mediated deimination of epithelial cell keratin results in cytoskeletal remodeling suggest a possible role for ePAD in cytoskeletal reorganization in the egg and early embryo. PMID- 12654295 TI - The midline (notochord and notoplate) patterns the cell motility underlying convergence and extension of the Xenopus neural plate. AB - We investigated the role of the dorsal midline structures, the notochord and notoplate, in patterning the cell motilities that underlie convergent extension of the Xenopus neural plate. In explants of deep neural plate with underlying dorsal mesoderm, lateral neural plate cells show a monopolar, medially directed protrusive activity. In contrast, neural plate explants lacking the underlying dorsal mesoderm show a bipolar, mediolaterally directed protrusive activity. Here, we report that "midlineless" explants consisting of the deep neural plate and underlying somitic mesoderm, but lacking a midline, show bipolar, mediolaterally oriented protrusive activity. Adding an ectopic midline to the lateral edge of these explants restores the monopolar protrusive activity over the entire extent of the midlineless explant. Monopolarized cells near the ectopic midline orient toward it, whereas those located near the original, removed midline orient toward this midline. This behavior can be explained by two signals emanating from the midline. We postulate that one signal polarizes neural plate deep cells and is labile and short-lived and that the second signal orients any polarized cells toward the midline and is persistent. PMID- 12654294 TI - An inducible system for the study of FGF signalling in early amphibian development. AB - The use of a novel inducible FGF signalling system in the frog Xenopus laevis is reported. We show that the lipophilic, synthetic, dimerizing agent AP20187 is able to rapidly activate signalling through an ectopically expressed mutant form of FGFR1 (iFGFR1) in Xenopus embryos. iFGFR1 lacks an extracellular ligand binding domain and contains an AP20187 binding domain fused to the intracellular domain of mouse FGFR1. Induction of signalling by AP20187 is possible until at least early neurula stages, and we demonstrate that ectopically expressed iFGFR1 protein persists until late neurula stages. We show that activation of signalling through iFGFR1 can mimic a number of previously reported FGF activities, including mesoderm induction, repression of anterior development, and neural posteriorization. We show that competence to morphological posteriorization of the anteroposterior axis by FGF signalling only extends until about stage 10.5. We demonstrate that the competence of neural tissue to express the posterior markers Hoxa7 and Xcad3, in response to FGF signalling, is lost by the end of gastrula stages. We also show that activation of FGF signalling stimulates morphogenetic movements in neural tissue until at least the end of the gastrula stage. PMID- 12654296 TI - Contribution of somitic cells to the avian ribs. AB - The traditional view that all parts of the ribs originate from the sclerotome of the thoracic somites has recently been challenged by an alternative view suggesting that only the proximal rib derives from the sclerotome, while the distal rib arises from regions of the dermomyotome. In view of this continuing controversy and to learn more about the cell interactions during rib morphogenesis, this study aimed to reveal the precise contributions made by somitic cells to the ribs and associated tissues of the thoracic cage. A replication-deficient lacZ-encoding retrovirus was utilized to label cell populations within distinct regions of somites 19-26 in stage 13-18 chick embryos. Analysis of the subsequent contributions made by these cells revealed that the thoracic somites are the sole source of cells for the ribs. More precisely, it is the sclerotome compartment of the somites that contributes cells to both the proximal and distal elements of the ribs, confirming the traditional view of the origin of the ribs. Results also indicate that the precursor cells of the ribs and intercostal muscles are intimately associated within the somite, a relationship that may be essential for proper rib morphogenesis. Finally, the data from this study also show that the distal ribs are largely subject to resegmentation, although cell mixing may occur at the most sternal extremities. PMID- 12654297 TI - Overexpression of transcription factor AP-2alpha suppresses mammary gland growth and morphogenesis. AB - AP-2 transcription factors are key regulators of mouse embryonic development. Aberrant expression of these genes has also been linked to the progression of human breast cancer. Here, we have investigated the role of the AP-2 gene family in the postnatal maturation of the mouse mammary gland. Analysis of AP-2 RNA and protein levels demonstrates that these genes are expressed in the mammary glands of virgin and pregnant mice. Subsequently, AP-2 expression declines during lactation and then is reactivated during involution. The AP-2alpha and AP-2gamma proteins are localized in the ductal epithelium, as well as in the terminal end buds, suggesting that they may influence growth of the ductal network. We have tested this hypothesis by targeting AP-2alpha expression to the mouse mammary gland using the MMTV promoter. Our studies indicate that overexpression of AP 2alpha inhibits mammary gland growth and morphogenesis, and this coincides with a rise in PTHrP expression. Alveolar budding is severely curtailed in transgenic virgin mice, while lobuloalveolar development and functional differentiation are inhibited during pregnancy and lactation, respectively. Our studies strongly support a role for the AP-2 proteins in regulating the proliferation and differentiation of mammary gland epithelial cells in both mouse and human. PMID- 12654298 TI - ADAM 10: an active metalloprotease expressed during avian epithelial morphogenesis. AB - The ADAMs are a family of proteins containing multiple functional domains. We have cloned the avian orthologue of ADAM 10 and demonstrate that it has metalloprotease activity. Chick ADAM 10 is expressed in the developing dermatome and myotome of the somite, epidermis, gut endoderm, the epithelial tissues of the kidney, liver, and heart, and in neural crest cells. The expression patterns and protein distribution of ADAM 10 suggest it may play a significant role in the morphogenesis of several epithelial tissues. When a dominant-negative metalloprotease-mutant form of ADAM 10 is expressed in the ectoderm or ADAM 10 expression is knocked down with morpholinos, morphogenesis and tissue specification are altered. PMID- 12654299 TI - Left-right patterning of the mouse lateral plate requires nodal produced in the node. AB - Initial determination of left-right (L-R) polarity in mammalian embryos takes place in the node. However, it is not known how asymmetric signals are generated in the node and transferred to the lateral plate mesoderm (LPM). Mice homozygous for a hypomorphic Nodal allele (Nodal(neo)) were generated and found to exhibit L R defects, including right isomerism. Although the mutant embryos express Nodal at gastrulation stages, the subsequent expression of this gene in the node and left LPM is lost. A transgene that conferred Nodal expression specifically in the node rescued the L-R defects of the Nodal(neo/neo) embryos. Conversely, ectopic expression of the Nodal inhibitor Lefty2 in the node of Nodal(neo/+) embryos resulted in a phenotype similar to that of the Nodal(neo/neo) mutant. These results indicate that Nodal produced in the node is required for expression of Nodal and other left side-specific genes in the LPM. PMID- 12654300 TI - UNC-52/perlecan affects gonadal leader cell migrations in C. elegans hermaphrodites through alterations in growth factor signaling. AB - The unc-52 gene of Claenorhabditis elegans encodes a homologue of the basement membrane heparan sulfate proteoglycan perlecan. Viable alleles reduce the abundance of UNC-52 in late larval stages and increase the frequency of distal tip cell (DTC) migration defects caused by mutations disrupting the UNC-6/netrin guidance system. These unc-52 alleles do not cause circumferential DTC migration defects in an otherwise wild-type genetic background. The effects of unc-52 mutations on DTC migrations are distinct from effects on myofilament organization and can be partially suppressed by mutations in several genes encoding growth factor-like molecules, including EGL-17/FGF, UNC-129/TGF-beta, DBL-1/TGF-beta, and EGL-20/WNT. We propose that UNC-52 serves dual roles in C. elegans larval development in the maintenance of muscle structure and the regulation of growth factor-like signaling pathways. PMID- 12654301 TI - Transgenic RNAi in mouse oocytes: a simple and fast approach to study gene function. AB - Double-strand RNA (dsRNA)-mediated posttranscriptional gene silencing, also known as RNA interference (RNAi), is a powerful tool to inhibit gene expression in several experimental model systems, including Arabidopsis, Caenorhabditis, and Drosophila. We previously described that the microinjection of Mos dsRNA into fully grown mouse oocytes results in the specific degradation of Mos mRNA in a time- and concentration-dependent manner. We report here a transgenic RNAi approach that is suitable to study gene function during mouse oocyte development and differentiation. The oocyte-specific Zp3 promoter was used to drive the expression of a long hairpin dsRNA ( approximately 500 bp) targeting Mos mRNA. Transgenic founder animals appeared healthy, but while males were fertile, females were not, in accordance with the known Mos null phenotype. The amount of Mos mRNA in the transgenic F(1) females was reduced by >90%, whereas there was no decrease in the nontargeted tissue plasminogen activator (Plat) mRNA. Moreover, the maturation-associated increase in mitogen-activated protein (MAP) kinase activity was not observed, and the metaphase II eggs underwent spontaneous parthenogenetic activation, thus recapitulating the Mos null phenotype. This approach provides a powerful method to study the functions of any oocyte synthesized gene during oocyte development and early embryogenesis. PMID- 12654302 TI - Measurement of UDP-N-acetylglucosaminyl transferase (OGT) in brain cytosol and characterization of anti-OGT antibodies. AB - UDP-N-acetylglucosaminyl transferase (OGT) catalyzes O-linked glycosylation of cytosolic and nuclear proteins, but enzyme studies have been hampered by the lack of a rapid, sensitive, and economical OGT assay. Employed assay methods typically involved the use of HPLC, formic acid, and large amounts of expensive radiolabeled [3H]UDP-N-acetylglucosaminyl ([3H]UDP-GlcNAc). In the current study, we have developed an OGT assay that circumvents many of these problems through four critical assay improvements: (1) identification of an abundant and enriched source of OGT enzyme (rat brain tissue), (2) utilization of a rapid method for efficiently removing salts and sugar nucleotides from cytosol (polyethylene glycol precipitation of active enzyme), (3) expression of a recombinant p62 acceptor substrate designed to facilitate purification (polyhistidine metal chelation site), and (4) development of two alternative methods to rapidly separate free [3H]UDP-GlcNAc from 3H-p62ST acceptor peptide (trichloroacetic acid precipitation and metal-chelation affinity purification). To study the enzymology of OGT, independent of potential regulatory proteins within cytosol, we also developed and characterized an alternate OGT assay that uses antibody-purified OGT as the enzyme source. The major advantage of this assay lies in the ability to measure OGT in the absence of other cytosolic proteins. PMID- 12654303 TI - Low-volume, high-sensitivity assay for cadmium in blood and urine using conventional atomic absorption spectrophotometry. AB - An assay for cadmium in whole blood and urine using deuterium background correction electrothermal atomic absorption spectroscopy (D(2)-ETAAS) was developed. Cadmium (in a 1- to 2-ml sample) was bound to 15 mg anion-exchange resin, interfering ions were removed in a 2-ml Bio-Spin column, and cadmium was extracted into 100 microl 1M nitric acid for analysis. Cadmium in the sample extract was concentrated 7-fold for blood and 10-fold for urine over the starting material. These steps produced cadmium atomic absorption traces with high signal to background ratios and allowed analysis against aqueous standards. At approximately 0.1 ng Cd/ml, mean intra- and interassay coefficients of variation were 11-12%. Cadmium recovery for 0.1 to 0.6 ng added cadmium was 107+/-4% for blood and 94+/-4% for urine (mean+/-SE, n=3). The mean detection limit (mean + 3 x SD of blank) was 0.008 ng/ml for blood and 0.003 ng/ml for urine. Samples from "unexposed" animals including humans ranged from 0.051+/-0.000 to 0.229+/-0.035 ng/ml. Values were approximately 10-fold lower than those obtained by the method of Stoeppler and Brandt using Zeeman background-correction ETAAS. This new high sensitivity, low-volume assay will be useful for epidemiological studies, even those involving children, and will provide a means to help determine the contribution of cadmium to disease incidence in the general population. PMID- 12654304 TI - Capillary electrophoresis of erythrocytes. AB - Capillary electrophoresis (CE) of erythrocytes from different sources under various conditions is reported in this paper. It was found that erythrocyte samples from sheep, duck, and human showed characteristic and reproducible elution peaks, and that the retention times of A-, B-, AB-, and O-type erythrocytes from human blood were distinctively different; even subtle differences, among individuals with the same blood type could be detected by CE. A strictly linear correlation was obtained between the peak area and the amount of human erythrocyte over a range of 4.8 x 10(2)-1.9 x 10(4) cells (r=0.999), indicating that CE could be used for rapid and accurate quantification of erythrocytes. Using this CE protocol, the decrease of the surface electrical charge of erythrocyte during storage was confirmed. Therefore, this work demonstrated that CE could be a useful alternative for characterizing and quantifying erythrocytes or other cells. PMID- 12654305 TI - Reagentless optical sensing of glutamine using a dual-emitting glutamine-binding protein. AB - Glutamine is a major source of nitrogen and carbon in cell culture media. Thus, glutamine monitoring is important in bioprocess control. Here we report a reagentless fluorescence sensing for glutamine based on the Escherichia coli glutamine-binding protein (GlnBP) that is sensitive in the submicromolar ranges. The S179C variant of GlnBP was labeled at the -SH and N-terminal positions with acrylodan and ruthenium bis-(2,2'-bipyridyl)-1,10-phenanthroline-9 isothiocyanate, respectively. The acrylodan emission is quenched in the presence of glutamine while the ruthenium acts as a nonresponsive long-lived reference. The apparent binding constant, K'(d), of 0.72 microM was calculated from the ratio of emission intensities of acrylodan and ruthenium (I(515)/I(610)). The presence of the long-lived ruthenium allowed for modulation sensing at lower frequencies (1-10 MHz) approaching an accuracy of +/-0.02 microM glutamine. Dual frequency ratiometric sensing was also demonstrated. Finally, the extraordinary sensitivity of GlnBP allows for dilution of the sample, thereby eliminating the effects of background fluorescence from the culture media. PMID- 12654306 TI - Novel bioluminescent assay of alkaline phosphatase using adenosine-3'-phosphate 5'-phosphosulfate as substrate and the luciferin-luciferase reaction and its application. AB - This paper describes a novel bioluminescent assay of alkaline phosphatase (ALP) utilizing ATP-sulfurylase and the luciferin-luciferase reaction. The principle governing the assay is as follows. Adenosine-3'-phosphate-5'-phosphosulfate, which serves as the substrate for ALP, is hydrolyzed enzymatically to produce adenosine-5'-phosphosulfate (APS). APS is converted into ATP by ATP-sulfurylase in the presence of pyrophosphate. The ATP produced is detected by the luciferin luciferase reaction. The measurable range was 1 zmol to 100 fmol/assay and the detection limit at blank+3 SD was 10 zmol/assay. The coefficient of variation (CV, n=5) was examined at each point of the standard curve; the mean CV percentage was 4.47% (n=6). This assay system was applied to enzyme immunoassay of human chorionic gonadotropin and allele-specific PCR enzyme-linked immunosorbent assay of verotoxin gene using ALP as the label enzyme; 10(-2) mIU/mL hCG in urine and 5 pg of Escherichia coli O157 DNA could be assayed directly and with high sensitivity by the proposed method. PMID- 12654307 TI - Polarographic direct determination and homogeneous immunoassay of anti-human serum albumin (HSA) by parallel catalytic hydrogen wave of anti-HSA or HSA. AB - Human serum albumin (HSA) or anti-human serum albumin (anti-HSA) yields a catalytic hydrogen wave at about -1.85V (vs Ag/AgCl) in 0.25M NH(3).H(2)O-NH(4)Cl (pH 8.58) buffer. When 1.0 x 10(-2)M K(2)S(2)O(8) is present, the catalytic hydrogen wave is further catalyzed, producing a parallel catalytic wave of hydrogen as catalyst in nature, termed the parallel catalytic hydrogen wave. The sensitivity of the parallel catalytic hydrogen wave is higher by two orders of magnitude than that of the catalytic hydrogen wave. Using the parallel catalytic hydrogen wave of anti-HSA or HSA in the presence of K(2)S(2)O(8), two sensitive methods for the determination of anti-HSA were developed. One is a direct determination based on the parallel catalytic hydrogen wave of anti-HAS itself, and the other is a homogeneous immunoassay based on measuring the decrease of the peak current of the parallel catalytic hydrogen wave of HSA after homogeneous immunoreaction of HSA with anti-HSA. In the direct determination, the second order derivative peak current of the parallel catalytic hydrogen wave of anti-HSA itself is rectilinear to its titer in the range from 1:1.0 x 10(7) to 1:8.4 x 10(6). In the homogeneous immunoassay, the decrease in the second-order derivative peak current of the parallel catalytic hydrogen wave of HSA is linearly related to the added anti-HSA in the titer range from 1:3.0 x 10(7) to 1:6.0 x 10(6). These assays are highly sensitive and rapid in operation and can be used to evaluate such antigens and their antibodies as those that could yield the parallel catalytic hydrogen wave. PMID- 12654308 TI - Molecular assay to generate expression profile of eight estrogen receptor alpha isoform mRNA copy numbers in picogram amounts of total RNA from breast cancer tissues. AB - The presence of estrogen receptor alpha (ERalpha) is the basis for treating breast cancer patients with targeted molecular therapies that block estrogen stimulation of breast cancer cell division. Currently, the ERalpha presence is determined by microscopically scoring the slides subjected to immunohistochemistry. This method is not quantitative, cannot distinguish between all the known ERalpha isoforms, and requires large amounts of tumor tissue. We describe here a real-time PCR-based molecular approach that can be applied to determine mRNA copies of eight ERalpha isoforms in picogram amounts of total RNA from clinical samples. Each isoform mRNA is quantified using a specific primer pair and a 5'FAM- and 3'TAMARA-labeled probe in comparison with a standard curve constructed with known copy numbers of its respective reverse-transcribed cRNA. Seven alternatively spliced isoforms were quantified using splice-targeted primers. The cRNAs for eight isoforms were generated by in vitro transcription of their respective coding sequences. The sensitivity of detection with reverse transcribed cRNAs is as low as 100 copies. The devised assays can detect ERalpha cDNAs reverse transcribed from as low as 50-100 pg of total RNA from breast cancer tissues. The applicability of the devised assays for profiling eight ERalpha isoform mRNAs is demonstrated using 6 breast cancer cell lines and 10 breast cancer tissues. It is expected that these assays could be applied to profile ERalpha isoforms in any estrogen-responsive tissues. In addition, these methods could highly facilitate the design of tissue-specific selective estrogen receptor modulators to treat breast cancers and other estrogen-related abnormalities. PMID- 12654309 TI - Isothermal titration calorimetric procedure to determine protein-metal ion binding parameters in the presence of excess metal ion or chelator. AB - Determination of binding parameters for metal ion binding to proteins usually requires preceding steps to remove protein-bound metal ions. Removal of bound metal ions from protein is often associated with decreased stability and inactivation. We present two simple isothermal titration calorimetric procedures that eliminate separate metal ion removal steps and directly monitor the exchange of metal ions between buffer, protein, and chelator. The concept is to add either excess chelator or metal ion to the protein under investigation and subsequently titrate with metal ion or chelator, respectively. It is thereby possible in the same experimental trial to obtain both chelator-metal ion and protein-metal ion binding parameters due to the different thermodynamic "fingerprints" of chelator and protein. The binding models and regression routines necessary to analyze the corresponding binding isotherms have been constructed. Verifications of the models have been done by titrations of mixtures of calcium chelators (BAPTA, HEDTA, and EGTA) and calcium ions and they were both able to account satisfactorily for the observed binding isotherms. Therefore, it was possible to determine stoichiometric and thermodynamic binding parameters. In addition, the concept has been tested on a recombinant alpha-amylase from Bacillus halmapalus where it proved to be a consistent procedure to obtain calcium binding parameters. PMID- 12654310 TI - Identification of the covalent flavin adenine dinucleotide-binding region in pyranose 2-oxidase from Trametes multicolor. AB - We present the first report on characterization of the covalent flavinylation site in flavoprotein pyranose 2-oxidase. Pyranose 2-oxidase from the basidiomycete fungus Trametes multicolor, catalyzing C-2/C-3 oxidation of several monosaccharides, shows typical absorption maxima of flavoproteins at 456, 345, and 275 nm. No release of flavin was observed after protein denaturation, indicating covalent attachment of the cofactor. The flavopeptide fragment resulting from tryptic/chymotryptic digestion of the purified enzyme was isolated by anion-exchange and reversed-phase high-performance liquid chromatography. The flavin type, attachment site, and mode of its linkage were determined by mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy of the intact flavopeptide, without its prior enzymatic degradation to the central aminoacyl moiety. Mass spectrometry identified the attached flavin as flavin adenine dinucleotide (FAD). Post-source decay analysis revealed that the flavin is covalently bound to histidine residue in the peptide STHW, consistent with the results of N-terminal amino acid sequencing by Edman degradation. The type of the aminoacyl flavin covalent link was determined by NMR spectroscopy, resulting in the structure 8alpha-(N(3)-histidyl)-FAD. PMID- 12654311 TI - Targeting the MraY and MurG bacterial enzymes for antimicrobial therapeutic intervention. AB - Assays for two enzymes from Escherichia coli were developed and validated as antibacterial inhibitor screens. The MraY and MurG enzymes were overexpressed and purified as the membrane fraction or to homogeneity, respectively. The MurG enzyme was expressed with a six-histidine tag using an optimized minimal-medium protocol for subsequent purification. Although traditional assays were established, the enzymes were also assayed via a 96-well membrane plate assay and a 384-well scintillation proximity-based assay developed herein. These assays afford a more economical and high-throughput evaluation of inhibitors. A mureidomycin inhibitor mix was used as a control for the assay development and screen validation. Several inhibitors resulting from a high-throughput screen were found and evaluated for potential therapeutic use. PMID- 12654313 TI - A microplate assay specific for the enzyme aggrecanase. AB - We have identified a 41-residue peptide, bracketing the aggrecanase cleavage site of aggrecan, that serves as a specific substrate for this enzyme family. Biotinylation of the peptide allowed its immobilization onto streptavidin-coated plates. Aggrecanase-mediated hydrolysis resulted in an immobilized product that reveals an N-terminal neoepitope, recognized by the specific antibody BC-3. This assay is highly specific for aggrecanases; MMPs were inactive in this assay. Reduction of the peptide size below 30 amino acids resulted in a significant diminution of activity. Using the immobilized 41-residue peptide as a substrate, we have developed a 96-well microplate-based assay that can be conveniently used for high-throughput screening of samples for aggrecanase activity and for discovery of inhibitors of aggrecanase activity. PMID- 12654312 TI - Series of vectors to evaluate the position and the order of two different affinity tags for purification of protein complexes. AB - A series of protein expression vectors with dual-affinity tags has been developed. With these constructed vectors, FLAG and hexahistidine tags were fused to a given protein at either the N- or the C-terminal ends or both, for a total of six combinations. Three auxotrophy markers were introduced into each construct, thus yielding 18 different vectors. These vectors allow evaluation of different positions and orders of two different tags. To confirm the efficacy of these vectors, we purified a histone acetyltransferase (Esa1p)-containing complex. First, an appropriate position of the tags was selected through small scale purification. Next, large-scale purification was done for the selected construct, yielding an Esa1p-containing complex that was comparable to an Esa1p containing complex (NuA4) obtained by a conventional activity-based purification. These vectors provide a convenient way to select the best position of tags for efficient purification of protein complexes also applicable in proteomics studies. PMID- 12654315 TI - Multivariate statistics in analysis of data from the in vitro motility assay. AB - A novel approach is described for classification of filaments as stationary or moving and for extraction of velocity data for smooth actin filament sliding in vitro. Moving and stationary filaments were effectively classified using four discriminating variables in a multivariate statistical analysis. The variables were (1) two different measures of the average filament distance from its starting point, (2) a measure of the variability in sliding direction, and (3) the coefficient of variation (CV) of the frame-to-frame sliding velocity (v(mean)). On the basis of this multivariate analysis we obtained correct classification of 98% of the stationary filaments and 94% of the moving filaments in a cross-validation data set. The same classification functions were useful throughout despite a 10-fold variation in the average sliding velocity in the cross-validation data. Further analysis of motile filaments suggested that the velocity of smooth sliding should, ideally, be obtained from the intercept on the velocity axis of a plot of v(mean) against CV. The velocity, so obtained, was between 10 and 30% (mean 20+/-3%; n=7; p<0.001) higher than if average sliding velocity was obtained for all moving filaments with CV<0.5. PMID- 12654314 TI - Site-specific characterization of the N-linked oligosaccharides of a murine immunoglobulin M by high-performance liquid chromatography/electrospray mass spectrometry. AB - Immunoglobulin M is an especially important product of the immune system because it plays a critical role in early protection against infections. In this report, the glycosylation pattern of the protective murine monoclonal IgM 12A1 to Cryptococcus neoformans polysaccharide was analyzed by high-performance liquid chromatography coupled with electrospray ionization mass spectrometry. Peptide mapping studies covering 88% of the deduced amino acid sequence indicated that of the six potential N-glycosylation sites in this antibody only five were utilized, as the tryptic peptide derived from monoclonal IgM 12A1 containing Asn-260 was recovered without carbohydrates. The oligosaccharide side chains of monoclonal IgM 12A1 were characterized at each of the N-glycosylation sites. Asn-166 possessed 20 monosialylated and nonsialylated, and fucosylated and nonfucosylated complex- and hybrid-type oligosaccharides and one high-mannose-type oligosaccharide. Thirteen oligosaccharides were attached to the site at Asn-401, including six complex-type, four hybrid-type, and three high-mannose-type oligosaccharides. Twelve hybrid-type oligosaccharides were attached to Asn-378, three of which had terminal sialic acids. Eleven hybrid-type oligosaccharides were attached to Asn-331, seven of which had terminal sialic acids. Only two high mannose type oligosaccharides were attached to Asn-363. These results indicated great complexity in the structure and composition of oligosaccharides attached to individual IgM glycosylation sites. PMID- 12654316 TI - Use of phage display and high-density screening for the isolation of an antibody against the 51-kDa subunit of complex I. AB - Monoclonal antibodies play an increasingly important role in structural biology. In this report, we develop the use of phage display technology for the isolation of an antibody that binds to a specific subunit of a macromolecular assembly. Antibodies that bind to the intact complex are selected from a phage display library and screened with a high-density Western blot to identify a subunit specific binder. Conventional Western blotting and competition ELISA are then used to confirm the identity of the target subunit and that the antibody binds to the native protein complex and not to an epitope that is only revealed when the antibody is immobilized for phage selection. Using this technique, monoclonal scFv and Fab fragments have been produced that bind to the 51-kDa subunit of bovine complex I, a large integral membrane protein complex from mitochondria. PMID- 12654318 TI - A gradient PCR-based screen for use in site-directed mutagenesis. AB - Site-directed mutagenesis is widely used to study protein and nucleic acid structure and function. Despite recent advancements in the efficiency of procedures for site-directed mutagenesis, the fraction of site-directed mutants by most procedures rarely exceeds 50% on a routine basis and is never 100%. Hence it is typically necessary to sequence two or three clones each time a site directed mutant is constructed. We describe a simple and robust gradient-PCR based screen for distinguishing site-directed mutants from the starting, unmutated plasmid. The procedure can use either purified plasmid DNA or colony PCR, starting from a single colony. The screen utilizes the primer used for mutagenesis and a common outside primer that can be used for all other mutants constructed with the same template. Over 30 site-specific mutants in a variety of templates were successfully screened and all of the mutations detected were subsequently confirmed by DNA sequencing. A single base pair mismatch could be detected in an oligonucleotide of 36 bases. Detection efficiency was relatively independent of starting template concentration and the nature of the outside primer used. PMID- 12654317 TI - Determination of affinities and antigenic epitopes of bovine cardiac troponin I (cTnI) with monoclonal antibodies by surface plasmon resonance biosensor. AB - A surface plasmon resonance (SPR) biosensor based on wavelength modulation was used for real-time detection of the interaction of three monoclonal antibodies and antigens of bovine cardiac troponin I (cTnI). In order to recognize antigenic epitopes of bovine cTnI, two experimental modes were applied. In the first experimental mode, three monoclonal antibodies were divided into three groups and three experiments were performed on biosensor surfaces prepared with protein A. In the second experimental mode, antigen was immobilized on the biosensor surface prepared by the amine-coupling method and three monoclonal antibodies were detected in turn. The results obtained by the two modes are consistent. In addition, the affinities of the monoclonal antibodies for the antigen were also determined by the association rate and the disassociation rate in real-time. These results validate the biosensor technology and illustrate how biosensors based on wavelength modulation can be used to study the interaction of monoclonal antibodies and antigens in real time. PMID- 12654319 TI - Transfecting mammalian cells by DNA/calcium phosphate precipitates: effect of temperature and pH on precipitation. PMID- 12654320 TI - RNA isolation from plant tissues rich in polysaccharides. PMID- 12654321 TI - Identity and localization of advanced glycation end products on human beta2 microglobulin using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. PMID- 12654323 TI - Prevalence and outcomes for parents with disabilities and their children in an Australian court sample. AB - OBJECTIVE: The purpose of this study was to obtain sound prevalence and outcomes data on parents with disabilities and their children in statutory child protection proceedings. METHOD: The court files of all care and protection matters initiated by the statutory child protection authority and finalized in a 9-month period (n=285) at two Children's Courts in NSW, Australia were reviewed. RESULTS: Parents with disabilities featured in almost one-third of the cases (29.5%). Parental psychiatric disability was most prevalent at 21.8% followed by parental intellectual disability (mental retardation) at 8.8%. Significant associations were found between parental disability and court outcome with a disproportionately large number of children of parents with intellectual disability being made wards of the state. CONCLUSIONS: The findings demonstrate that parents with disabilities are significantly over-represented in statutory child protection proceedings and that the outcomes of these proceedings vary according to disability type. PMID- 12654322 TI - Parents with disabilities involved in dependency cases. PMID- 12654324 TI - Street children and political violence: a socio-demographic analysis of street children in Rwanda. AB - OBJECTIVE: The aims were: (1) to examine the profile of African street children and to assess the link between street children in Africa and political violence; (2) to undertake a systematic examination of causal factors of street children in postgenocide Rwanda; and (3) to situate this analysis in the context of the socio cultural and political impact of the genocide on Rwandan communities. METHOD: Observational mapping examined the profile and activities of Rwandan street children. Structured interviews were carried out with 290 children in four regional towns to obtain information on socio-demographic, familial, educational background, causal factors surrounding street life involvement, psychological well-being, and relationship to the street. Focus group discussions and key informant interviews examined the relationship between street children and the broader Rwandan society. RESULTS: Street children in Rwanda were predominantly adolescent boys, almost half of whom were homeless (42%), with a high proportion of orphaned children or children who had lost at least one parent. Two variables predicted homelessness: child's guardian and reason for being in street. Qualitative accounts of children conveyed the impact of death of family members, repatriation, imprisonment of parents, and poverty on their lives. CONCLUSIONS: The analysis highlighted the need for community based support for children in alternative guardianship care and for policies to support the reintegration of male youths in postconflict welfare strategies as prevention strategies for street migration. PMID- 12654325 TI - Abusive head trauma in Maine infants: medical, child protective, and law enforcement analysis. AB - OBJECTIVE: To collect and compare the results of medical, child protective, and law enforcement evaluation of a sample of Maine children who were victims of abusive head trauma (AHT) in order to describe the clinical and evaluative characteristics as they relate to victims, families and perpetrators of such trauma and to improve the professional response to AHT in Maine. METHOD: Retrospective chart review of medical, child protective, and law enforcement records of all AHT victims admitted to two tertiary care hospitals in Maine or seen by the state medical examiner from 1991 to 1994. RESULTS: Nineteen children (age range 2 weeks to 17 months) were identified as victims of AHT (out of a total of 94 head trauma admissions) accounting for 20 hospitalizations during the study period. There was a history of prior injury in 30%, history of prior medical evaluations for possibly abuse related problems in 65%, while, on presentation, 75% had evidence or history of prior injury. The hospitals notified child protective services (CPS) in all 20 cases and correctly identified abuse in 18 (90%). Parental risk factors for abuse identified in CPS records included substance abuse (53%), domestic violence (42%), criminal history (32%), unrealistic expectations (42%), and attachment problems (32%). However, risk factors were inadequately assessed in 53% of homes. Law enforcement identified a likely perpetrator in 79% of cases and in the majority the identified suspect was the father. In the 15 cases where a perpetrator was identified by law enforcement, that person was alone with the child at symptom onset in 14 (93%). CONCLUSIONS: The medical response, at least at the inpatient level, was generally well done with regard to suspicion and reporting. Cases are possibly being missed at the outpatient level. Child protective risk assessment was limited overall yet in a third of the homes where AHT occurred, few if any risk factors were present to aid in identification and prevention. Law enforcement results suggest that a primary suspect for AHT is the caretaker alone with the child at the time of symptom onset. PMID- 12654326 TI - Interpretations of child compliance in individuals at high- and low-risk for child physical abuse. AB - OBJECTIVE: Our studies compared individuals at high- and low-risk for child physical abuse on measures of social information processing. METHOD: Two studies were conducted using similar methods. Twenty-eight childless women in Study 1 and 36 mothers in Study 2 read vignettes of parent-child interactions in which the child's level of compliance was difficult to interpret. Participants were asked a series of questions about the child's behavior and their own reactions. RESULTS: Accuracy and bias in identifying compliant behavior were assessed using a signal detection paradigm. In both samples, high- and low-risk participants did not differ in their overall accuracy in identifying children's behaviors. However, they used different evaluation standards such that high-risk participants were biased toward seeing more noncompliance and low-risk participants were biased toward seeing more compliance. High- and low-risk participants also made different types of errors in interpreting children's behavior. Low-risk participants were more likely to misinterpret noncompliant behavior as compliant, and there was a trend for high-risk participants to not perceive compliant behavior when it occurred. There were no differences in reported disciplinary responses in either study and the results for affective reactions were mixed. CONCLUSIONS: Specific differences in social information processing between high- and low-risk individuals replicated across samples, suggesting a reliable association between evaluation standards and risk of child physical abuse. However, the absence of differences in reported discipline and inconsistent findings on affective reactions indicate the need to identify the mechanism through which cognition influences parenting behavior. PMID- 12654327 TI - A comparison of pregnant women presenting for alcohol and other drug treatment by CPS status. AB - OBJECTIVE: The purpose of this study was to describe the demographic, substance use, and treatment variables of alcohol and other drug (AOD) treatment seeking pregnant women. This study also compared women on these variables based on Child Protective Service (CPS) status. METHOD: Secondary data of all pregnant women with at least one child entering all publicly funded AOD treatment programs (N=678), in a southwest county, over a 3-year period were analyzed to examine the differences between pregnant women also involved with CPS and those not involved with CPS. RESULTS: Pregnant women reporting CPS involvement were similar to non CPS women on demographic variables but differed on drug use and treatment variables. CPS women were more likely to report marijuana use as their primary problem drug, be mandated to treatment, attend day treatment and be released from treatment unsatisfactorily compared to the non-CPS pregnant women. Those without CPS involvement were more likely to report cocaine or crack as their primary drug, attend outpatient treatment and be found to have a satisfactory release from treatment compared to those with CPS involvement. Results from the multiple logistic regression analysis found that the significant predictors of CPS involvement were mandated status and unsatisfactory treatment release. CONCLUSIONS: Differences in treatment outcomes may be based on AOD agencies' difficulty in working with mandated clients and/or with the multiple demands placed by CPS on clients as part of their treatment plans. Further research is needed to determine reasons for CPS clients' high failure rates in AOD treatment. PMID- 12654328 TI - When is the personal professional in public child welfare practice? The influence of intimate partner and child abuse histories on workers in domestic violence cases. AB - OBJECTIVE: The objective of this article is to examine children's services workers' (CSWs') personal histories of abuse and their influence on professional responses to allegations of domestic violence. METHOD: Using an anonymous questionnaire, data were collected from CSWs in two large urban counties in Southern California who participated in a domestic violence training program (n=303). It was hypothesized that CSWs' responses to domestic violence cases would be affected by personal histories of abuse and the degree to which the CSWs identified with battered women. RESULTS: Approximately one-half of the respondents reported experiencing physical and/or sexual violence by an intimate partner. One-third of respondents reported physical abuse, and 22%, sexual abuse during childhood. The experience of childhood sexual abuse, especially for female CSWs, was associated with increased support for removal of children whose mother is being abused. Those CSWs with a history of partner violence who identified with battered women were less likely to approve of removing children from the battered mother than were CSWs without partner abuse history or identification with battered women. CONCLUSIONS: Our findings provide support for expanded training efforts that recognize the ongoing impact of victimization on CSWs' professional functioning. PMID- 12654329 TI - Child maltreatment in the "children of the nineties:" the role of the child. AB - AIM: To determine characteristics of children that may predispose to maltreatment. METHODS: The research is based on a large cohort study, the Avon Longitudinal Study of Parents and Children. Out of 14,256 children participating in the study, 115 have been identified as having been placed on local child protection registers prior to their 6th birthday. Data on the children have been obtained from obstetric data and from a series of parental questionnaires administered during pregnancy and the first 3 years of life. Risk factors have been analysed using logistic regression analysis. RESULTS: Significant relationships were found between low birthweight (OR 2.08), unintended pregnancies (OR 2.92), poor health (OR 1.91) and developmental problems (OR 1.99) in infancy, and subsequent maltreatment. In addition, mothers of registered children were less likely to have reported positive attributes in their 4-week old infant. In contrast, negative attributes in infancy, feeding and crying problems, and frequent temper tantrums were not significantly associated with maltreatment. CONCLUSIONS: While child factors are significant, they are only a small part of the overall complex set of circumstances and conditions that ultimately lead to abuse or neglect. Parental attitudes towards the child may be more significant than the actual characteristics of the child. PMID- 12654330 TI - Similar planning strategies for whole-body and arm movements performed in the sagittal plane. AB - The present paper looks for kinematic similarities between whole-body and arm movements executed in the sagittal plane. Eight subjects performed sit-to-stand (STS) and back-to-sit (BTS) movements at their preferred speed in the sagittal plane. Kinematics analysis focused on shoulder motion revealed that STS was composed of a straight, forward displacement followed by a curved, upward displacement while BTS was characterized by a curved, downward and straight, backward displacement. Curvature of the upward displacement was significantly greater than the downward one. Analysis of shoulder-velocity profiles showed that movement duration was significantly longer for BTS compared with STS and that the shape of the velocity profiles changed when subjects performed an STS compared with a BTS movement. Velocity profiles of the upward and downward displacements also differed; the relative acceleration duration (acceleration duration divided by movement duration during the vertical motion) was smaller for the upward compared with the downward displacement. The present results are in accordance with previous findings concerning the execution of vertical arm movements and suggest that the CNS uses similar motor plans for the performance of arm and whole-body movements in the sagittal plane. PMID- 12654331 TI - Increase of hippocampal acetylcholine release at the onset of dark phase is suppressed in a mutant mice model of evening-type individuals. AB - We have previously reported that clock mutant mice on Jcl:ICR background show about 2-h delayed circadian profiles in body temperature, spontaneous activity and sleep-wake rhythm, and that they appear to be useful as a model of evening type of individual. Hippocampal acetylcholine (ACh) release which is positively correlated with attention, learning and memory shows a circadian variation. In this study, changes in hippocampal ACh release in transitional phase from light (rest) to dark (active) period in clock mutant mice were monitored using an in vivo microdialysis method. Compared with wild mice, the increase in hippocampal ACh in the first 2 h of the active period in the mutant mice was suppressed in parallel with peak frequency in electroencephalogram theta rhythm. The molecular basis of the circadian system appears to have a strong effect on hippocampal cholinergic function, and is probably associating with individual temporal differences in voluntary behavior, cognition, learning and/or memory performance. PMID- 12654332 TI - Estradiol affects axo-somatic contacts of neuroendocrine cells in the arcuate nucleus of adult rats. AB - It has been shown that gonadal steroids have the capacity to induce synaptic plasticity in certain areas of the nervous system. Previously we have demonstrated that due to the effect of estradiol there is a transient decrease in the number of GABAergic axo-somatic synapses in the arcuate nucleus. By using systemic application of the tracer Fluorogold we retrogradely labeled a subpopulation of arcuate neurons that project to the median eminence. We than applied the disector method for synapse quantification and found that these "hypophysiotropic neurons" receive less axo-somatic inputs. We found that 17beta estradiol induced a decrease in the numerical density of axo-somatic contacts of these retrogradely-labeled neoroendocrine cells. Our data support the hypothesis that the hormonally driven morphological synaptic plasticity is neuron specific within the arcuate nucleus and plays a decisive role in the regulation of anterior pituitary. PMID- 12654333 TI - Galanin expression in adult human dorsal root ganglion neurons: initial observations. AB - Human dorsal root ganglia (DRGs) were obtained during various procedures and processed for single and double in situ hybridisation using oligonucleotide probes complementary to three peptide mRNAs. Some postmortem ganglia were also analysed. In donor (unlesioned) DRGs 12.5% of the neuron profiles (NPs) were galanin mRNA-positive (mRNA(+)), 47.5% calcitonin gene-related peptide (CGRP) mRNA(+) and 32.7% substance P mRNA(+). The corresponding percentages for cervical/thoracic DRGs from patients suffering from severe brachial plexus injury were 32.8%, 57.4% and 34.5%, respectively. In these DRGs a high proportion of the galanin mRNA(+) NPs contained CGRP mRNA and substance P mRNA. In DRGs from a patient with migraine-like pain a comparatively small proportion expressed galanin, whereas in DRGs from a herpes zoster patient galanin mRNA(+) NPs were comparatively more frequent. The results from human postmortem DRGs revealed only weak peptide mRNA signals. The present results demonstrate that galanin is expressed in DRGs not only in a number of animal species including monkey as previously shown, but also in a considerable proportion of human DRG neurons, often together with CGRP and substance P, and mostly in small neurons. Thus, galanin may play a role in processing of sensory information, especially pain, in human DRGs and dorsal horn. However, to what extent a similarly dramatic upregulation of galanin expression can be seen after peripheral nerve lesion in man, as has been reported for rat, mouse and monkey, remains to be analysed. PMID- 12654335 TI - Long-term potentiation and long-term depression in hippocampal CA1 neurons of mice lacking the IP(3) type 1 receptor. AB - To investigate the role in synaptic plasticity of Ca(2+) released from intracellular Ca(2+) stores, mice lacking the inositol 1,4,5-trisphosphate type 1 receptor were developed and the physiological properties, long-term potentiation, and long-term depression of their hippocampal CA1 neurons were examined. There were no significant differences in basic synaptic functions, such as membrane properties and the input/output relationship, between homozygote mutant and wild type mice. Enhanced paired-pulse facilitation at interpulse intervals of less than 60 ms and enhanced post-tetanic potentiation were observed in the mutant mice, suggesting that the presynaptic mechanism was altered by the absence of the inositol 1,4,5-trisphosphate type 1 receptor. Long-term potentiation in the field excitatory postsynaptic potentials induced by tetanus (100 Hz, 1 s) and the excitatory postsynaptic currents induced by paired stimulation in hippocampal CA1 pyramidal neurons under whole-cell clamp conditions were significantly greater in mutant mice than in wild-type mice. Homosynaptic long-term depression of CA1 synaptic responses induced by low-frequency stimulation (1 Hz, 500 pulses) was not significantly different, but heterosynaptic depression of the non-associated pathway induced by tetanus was blocked in the mutant mice. Both long-term potentiation and long-term depression in mutant mice were completely dependent on N-methyl-D-aspartate receptor activity. To rule out the possibility of an effect compensating for the lack of the inositol 1,4,5-trisphosphate type 1 receptor occurring during development, an anti-inositol 1,4,5-trisphosphate type 1 receptor monoclonal antibody that blocks receptor function was diffused into the wild-type cell through a patch pipette, and the effect of acute block of inositol 1,4,5-trisphosphate type 1 receptor on long-term potentiation was examined. Significant enhancement of long-term potentiation was observed compared with after control immunoglobulin G injection, suggesting that developmental redundancy was not responsible for the increase in long-term potentiation amplitude observed in the mutant mouse. The properties of channels that could be involved in long-term potentiation induction were examined using whole-cell recording. N-methyl-D-aspartate currents were significantly larger in mutant mice than in wild-type mice only between holding potentials of -60 and -80 mV. We conclude that inositol 1,4,5-trisphosphate type 1 receptor activity is not essential for the induction of synaptic plasticity in hippocampal CA1 neurons, but appears to negatively regulate long-term potentiation induction by mild modulation of channel activities. PMID- 12654334 TI - 5'-aminoimidazole-4-carboxamide riboside induces apoptosis in human neuroblastoma cells. AB - 5'-Aminoimidazole-4-carboxamide riboside (AICA riboside) has been previously shown to be toxic to two neuronal cell models [Neuroreport 11 (2000) 1827]. In this paper we demonstrate that AICA riboside promotes apoptosis in undifferentiated human neuroblastoma cells (SH-SY5Y), inducing a raise in caspase 3 activity. In order to exert its effect on viability, AICA riboside must enter the cells and be phosphorylated to the ribotide, since both a nucleoside transport inhibitor, and an inhibitor of adenosine kinase produce an enhancement of the viability of AICA riboside-treated cells. Short-term incubations (2 h) with AICA riboside result in five-fold increase in the activity of AMP-dependent protein kinase (AMPK). However, the activity of AMPK is not significantly affected at prolonged incubations (48 h), when the apoptotic effect of AICA riboside is evident. The results demonstrate that when the cell line is induced to differentiate both toward a cholinergic phenotype (with retinoic acid) or a noradrenergic phenotype (with phorbol esters), the toxic effect is significantly reduced, and in the case of the noradrenergic phenotype differentiation, the riboside is completely ineffective in promoting apoptosis. This reduction of effect correlates with an overexpression of Bcl-2 during differentiation. AICA riboside, derived from the hydrolysis of the ribotide, an intermediate of purine de novo synthesis, is absent in normal healthy cells; however it may accumulate in those individuals in which an inborn error of purine metabolism causes an increase in the rate of de novo synthesis and/or an overexpression of cytosolic 5'-nucleotidase, that appears to be the enzyme responsible for AICA ribotide hydrolysis. In fact, 5'-nucleotidase activity has been shown to increase in patients affected by Lesch-Nyhan syndrome in which both acceleration of de novo synthesis and accumulation of AICA ribotide has been described, and also in other neurological disorders of unknown etiology. Our results raise the intriguing clue that the neurotoxic effect of AICA riboside on the developing brain might contribute to the neurological manifestations of syndromes related to purine dismetabolisms. PMID- 12654336 TI - Tracing developing pathways in the brain: a comparison of carbocyanine dyes and cholera toxin b subunit. AB - The present study examined the efficiency of fluorescent carbocyanine dye 1,1' dioctadecyl-3,3,3',3'-tetramethylinodocarbocyanine perchlorate and cholera toxin B subunit in tracing the crossed tectal projection to the nucleus rotundus of the thalamus (tectorotundal pathways) of paraformaldehyde-fixed and living chick embryos. The tracers were injected into the optic tectum under three experimental conditions (carbocyanine postfix, carbocyanine in vivo, and cholera toxin B subunit in vivo) and the anterograde transport of the nucleus rotundus was monitored and compared. In the carbocyanine postfix method, small crystals of carbocyanine dye were inserted into the tectum of paraformaldehyde-fixed embryos. A 6-month post-insertion period was required to label the crossed tectorotundal pathway. Results showed that tectal neurons did not begin to innervate the ipsilateral nucleus rotundus until embryonic day 9 and the contralateral nucleus rotundus until embryonic day 17. This slow progression of labeling through the crossed tectal projection resulted in significant contrast of the labeling between the ipsilateral and contralateral nuclei rotundus. In the carbocyanine in vivo method, a small volume of carbocyanine dye solution was injected into the tectum of living embryos. A 8- to 12-h survival period was sufficient enough to label the tectorotundal pathway. By embryonic day 8, the labeled axons terminated in the ipsilateral nucleus rotundus and the crossed tectorotundal projection was first detected by embryonic day 10. Similarly, in the cholera toxin B subunit in vivo method, a small volume of cholera toxin B subunit solution was injected into the tectum of living embryos. After a 6- to 10-h survival period, heavily labeled axons were found to innervate bilaterally the nucleus rotundus by embryonic day 8. This appeared to be the earliest schedule for detecting the crossed tectorotundal projection, compared with that of both the postfix and in vivo methods of carbocyanine dye. Based on the differences in the detectability of the crossed tectorotundal projection between the postfix and in vivo methods, the present data suggest that the former method is of limited purpose for labeling tectal collaterals during embryogenesis. Moreover, given the rapid transport rate and absence of photobleaching, which is often seen when using carbocyanine dye, the cholera toxin B subunit in vivo method appears to be the tracer of choice for investigating embryonic pathways. PMID- 12654337 TI - In vivo role of truncated trkb receptors during sensory ganglion neurogenesis. AB - The mammalian trkB locus undergoes alternative splicing to produce two different types of brain-derived neurotrophic factor receptors. The first type is the full length receptor tyrosine kinase (TrkB(Tk+); the second type is a truncated receptor lacking the intracellular tyrosine kinase domain (TrkB(Tk-)). To investigate the function of both types of TrkB receptor in vivo, we have generated knockout mice lacking all isoforms of the TrkB receptor (trkB-/-) and compared sensory neuron survival in these mice to that in the previously described TrkB kinase domain knockout mice (trkB(k)-/-). We observed that the presence of truncated TrkB receptors in trkB(k)-/- mice results in more severe sensory neuron losses. Increased neuron losses associated with the presence of truncated TrkB were most severe in regions where neuron survival is most dependent on brain-derived neurotrophic factor and neurotrophin-3. Our data suggest that truncated TrkB receptors negatively influence neuron survival by interfering with the function of catalytic TrkB receptors. PMID- 12654338 TI - Molecular mechanisms of neuronal death in the dorsal lateral geniculate nucleus following visual cortical lesions. AB - We investigated the molecular mechanisms of cell death in the dorsal lateral geniculate nucleus of the rat, following suction lesion of the visual cortex at birth or in the third postnatal week, using terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling (TUNEL) technique and immunohistochemistry for caspase-3, -7, -8, and cleaved poly(ADP-ribose) polymerase. Following lesion at birth, TUNEL-positive neurons were found in the dorsal lateral geniculate nucleus between 24 h and 3 days after lesion, with a peak on the second day. Shorter survival times (12-18 h) resulted in labeling of very few neurons in dorsal lateral geniculate nucleus and of several neurons in the perilesional cortex. Activated caspase-3 was expressed from the first to the third days after lesion, whereas cleaved poly(ADP-ribose) polymerase and activated caspase-8 were expressed on the second and third day. Activated caspase-7 was expressed mainly in pretectal nuclei. Caspase-3 activation coincided with the appearance of TUNEL positive profiles, but decreased earlier than TUNEL. In the ipsi- and contralateral cerebral cortex, all parameters were unchanged. In animals lesioned in the third week, rare apoptotic thalamic neurons were detected as TUNEL- and activated caspase-3-positive profiles 2 days after cortical ablation, and were still present 1 week after lesion.Thus, early target ablation has dramatic effects on neonatal thalamic neurons, which die following activation of caspases 3 and 8. In contrast, cortical neurons are relatively unaffected by target deprivation. Compared with early lesions, late lesions induce a limited thalamic cell death, that persists over time. PMID- 12654339 TI - Chronic psychosocial stress decreases calcineurin in the dentate gyrus: a possible mechanism for preservation of early ltp. AB - Chronic psychosocial stress impairs early long-term potentiation (LTP) in the hippocampal CA1 region but not in the dentate gyrus of anesthetized rats. Analysis of putative signaling molecules involved in the expression of LTP was performed to determine the possible reason(s) for the apparent resistance of the LTP of the dentate gyrus to chronic psychosocial stress. Immunoblotting was used to determine possible changes in the basal levels of various fractions of calcium dependent calmodulin kinase II (CaMKII), phosphorylated CaMKII (P-CaMKII), calmodulin, protein kinase C gamma (PKCgamma) and calcineurin in the dentate gyrus of chronically stressed rats. Western blot analysis revealed that chronic stress significantly decreased the levels of the total CaMKII without affecting P CaMKII levels. No significant change was detected in the levels of the upstream effectors, calmodulin and PKCgamma. However, chronic stress produced a significant decrease in calcineurin levels. The data suggest that the dentate gyrus of chronically stressed rats may have developed a compensatory mechanism whereby calcineurin levels are reduced to maintain normal P-CaMKII levels, which may be responsible for the normal early LTP of the dentate gyrus of chronically stressed rats. The results of this work will increase understanding of why certain brain regions are more resistant to deleterious effects of conditions that deteriorate learning and memory. PMID- 12654340 TI - Regional distribution of alpha(2C)-adrenoceptors in brain and spinal cord of control mice and transgenic mice overexpressing the alpha(2C)-subtype: an autoradiographic study with [(3)H]RX821002 and [(3)H]rauwolscine. AB - Behavioral studies on gene-manipulated mice have started to elucidate the neurobiological functions of the alpha(2C)-adrenoceptor (AR) subtype. In this study, we applied quantitative receptor autoradiography to investigate the potential anatomical correlates of the observed functional effects of altered alpha(2C)-AR expression. Labeling of brain and spinal cord sections with the subtype non-selective alpha(2)-AR radioligand [(3)H]RX821002 and the alpha(2C)-AR preferring ligand [(3)H]rauwolscine revealed distinct binding-site distribution patterns. In control mice, [(3)H]rauwolscine binding was most abundant in the olfactory tubercle, accumbens and caudate putamen nuclei, and in the CA1 field of the hippocampus. A mouse strain with overexpression of alpha(2C)-AR regulated by a gene-specific promoter showed approximately two- to four-fold increased levels of [(3)H]rauwolscine binding in these regions. In addition, dramatic increases in [(3)H]rauwolscine binding were seen in the nerve layer of the olfactory bulb, the molecular layer of the cerebellum, and the ventricular system of alpha(2C)-AR overexpressing mice, representing "ectopic" alpha(2C)-AR expression. Competition binding experiments with several alpha(2)-AR ligands confirmed the alpha(2C)-AR identity of these sites. Our results provide quantitative evidence of the predominance of the alpha(2A)-AR subtype in most regions of the mouse CNS, but also disclose the wide distribution of alpha(2C)-AR in the normal mouse brain, although at relatively low density, except in the ventral and dorsal striatum and the hippocampal CA1 area. alpha(2C)-AR are thus present in brain regions involved in the processing of sensory information and in the control of motor and emotion related activities such as the accumbens and caudate putamen nuclei, the olfactory tubercle, the lateral septum, the hippocampus, the amygdala, and the frontal and somatosensory cortices. The current results may help in specifying an anatomical framework for the functional roles of the alpha(2A)- and alpha(2C)-AR subtypes in the mouse CNS. PMID- 12654341 TI - Ammonia prevents glutamate-induced but not low K(+)-induced apoptosis in cerebellar neurons in culture. AB - Cultured rat cerebellar granule neurons are widely used as a model system for studying neuronal apoptosis. Either low K(+) (5 mM) or low concentrations of glutamate (1-10 microM) induce apoptosis in cerebellar neurons in culture. However, the molecular mechanism(s) involved remain unclear. We show that long term treatment with ammonia prevents glutamate-induced but not low K(+)-induced apoptosis in cerebellar neurons, as assessed by measuring DNA fragmentation and activation of caspase 3. Ammonia prevented glutamate-induced increase of intracellular calcium, depolarization of the inner mitochondrial membrane, release of cytochrome c to the cytosol, activation of caspase 3 and fragmentation of DNA. However, ammonia did not prevent low K(+)-induced activation of caspase 3 and fragmentation of DNA. These results indicate that the initial steps involved in the induction of apoptosis by low K(+) or by glutamate are different and that ammonia prevents glutamate-induced apoptosis by reducing glutamate-induced rise of intracellular Ca(2+), thus avoiding the activation of subsequent events of the apoptotic process. PMID- 12654344 TI - Basal ganglia and processing of cortical information: functional interactions between trans-striatal and trans-subthalamic circuits in the substantia nigra pars reticulata. AB - The substantia nigra pars reticulata (SNR), a major output station of basal ganglia, receives information from the cerebral cortex through three main pathways, i.e. a direct inhibitory trans-striatal pathway, an indirect excitatory trans-striatal pathway that involves the pallidum and the subthalamus and a direct excitatory trans-subthalamic pathway. In order to determine how cortical information flow originating from functionally distinct cortical areas and processed through the trans-striatal and trans-subthalamic pathways is integrated within the SNR, the responses induced by electrical stimulation of prefrontal, motor and auditory cortex in SNR cells were analyzed in anesthetized rats. Further confirming that direct striato-nigral pathways related to these functionally distinct cortical areas are organized in parallel channels, stimulation of the prefrontal, motor and auditory cortex induced an inhibitory response on distinct subpopulations of SNR cells. Within a given channel, the direct trans-striatal and the trans-subthalamic pathways converge on a large number of nigral cells. In addition, the present study reveals that nigral cells receiving an inhibitory input from a given cortical area through the direct trans striatal pathway can also receive an excitatory input from a functionally distinct cortical area through the trans-subthalamic pathways. Such a convergence mainly occurred between the direct striato-nigral pathway issued from the auditory cortex and the trans-subthalamic pathways issued from the motor cortex. These data reveal the existence of a converging influence of trans-subthalamic and direct striato-nigral pathways not only within but also across channels. Within a given cortico-basal ganglia channel, the trans-subthalamic pathways likely contribute to the temporal shaping of the striato-nigral inhibition and thus of the disinhibition of the related nigral target nuclei in the thalamus and mesencephalon. Across channels, the specific interactions between trans subthalamic and direct striato-nigral pathways could contribute to prevent inhibition of subpopulations of nigral cells implicated in competing functions. PMID- 12654342 TI - The selective cyclooxygenase-2 inhibitor rofecoxib suppresses brain inflammation and protects cholinergic neurons from excitotoxic degeneration in vivo. AB - Brain inflammatory processes underlie the pathogenesis of Alzheimer's disease, and non-steroidal anti-inflammatory drugs have a protective effect in the disease. The aim of this work was to study in vivo whether attenuation of brain inflammatory response to excitotoxic insult by the selective cyclooxygenase-2 inhibitor, rofecoxib, may prevent neurodegeneration, as a contribution to a better understanding of the role inflammation plays in the pathology of Alzheimer's disease. We investigated, by immunohistochemical methods, glia reaction, the activation of p38 mitogen-activated protein kinase (p38MAPK) pathway with an antibody selective for the phosphorylated form of the enzyme and the number of choline acetyltransferase-positive neurons and, by in vivo microdialysis, cortical extracellular levels of acetylcholine following the injection of quisqualic acid into the right nucleus basalis of adult rats. Seven days after injection, a marked reduction in the number of choline acetyltransferase-positive neurons was found, along with an intense glia reaction, selective activation of p38MAPK at the injection site and a significant decrease in the extracellular levels of acetylcholine in the cortex ipsilateral to the injection site. The loss of cholinergic neurons persisted for at least up to 28 days. Rofecoxib (3 mg/kg/day, starting 1 h prior to injection of quisqualic acid) treatment for 7 days significantly attenuated glia activation and prevented the loss of choline acetyltransferase-positive cells and a decrease in cortical acetylcholine release. The prevention of cholinergic cell loss by rofecoxib occurred concomitantly with the inhibition of p38MAPK phosphorylation. Our findings suggest an important role of brain inflammatory reaction in cholinergic degeneration and demonstrate a neuroprotective effect of rofecoxib, presumably mediated through the inhibition of p38MAPK phosphorylation. PMID- 12654343 TI - Mu opioid receptor signaling in morphine sensitization. AB - We used a previously reported model of morphine sensitization that elicited a complex behavioral syndrome involving stereotyped and non stereotyped activity. To identify the mechanism of these long-lasting processes, we checked the density of mu opioid receptors, receptor-G-protein coupling and the cyclic AMP (cAMP) cascade. In morphine-sensitized animals mu opioid receptor autoradiography revealed a significant increase in the caudate putamen (30% versus controls), nucleus accumbens shell (16%), prefrontal and frontal cortex (26%), medial thalamus (43%), hypothalamus (200%) and central gray (89%). Concerning morphine's activation of G proteins in the brain, investigated in the guanylyl 5'-[gamma (35)S]thio]triphosphate ([(35)S]GTPgammaS) binding assay, a significant increase in net [(35)S]GTPgammaS binding was seen in the caudate putamen (39%) and hypothalamus (27%). In the caudate putamen this was due to an increase in the amount of activated G proteins, and in the hypothalamus to a greater affinity of G proteins for guanosine triphosphate (GTP). The main second messenger system linked to the opioid receptor is the cAMP pathway. In the striatum basal cAMP levels were significantly elevated in sensitized animals (70% versus controls) and [D-Ala(2), N-Me-Phe(4), Gly(5)-ol]-enkephalin (DAMGO) significantly inhibited forskolin-stimulated cAMP production in control (30%) but not in sensitized rats. In the hypothalamus no significant changes were observed in basal cAMP levels and DAMGO inhibition. These cellular events induced by morphine pre-exposure could underlie the neuroadaptive processes involved in morphine sensitization. PMID- 12654346 TI - Attenuation of N-methyl-D-aspartate-mediated cytoplasmic vacuolization in primary rat hippocampal neurons by mood stabilizers. AB - Recent post-mortem and brain imaging studies suggest that decreased neuronal and glial densities may account for cell loss in vulnerable brain regions such as the hippocampus and the frontal cortex in patients with bipolar disorder. Investigations into the mechanisms of action of mood stabilizers suggest that these drugs may regulate the expression of neuroprotective genes and protect against excitotoxicity. In this study, we characterized the ultrastructural appearance of rat hippocampal neurons pretreated with mood stabilizers and then exposed to the glutamate receptor agonist N-methyl-D-aspartate. Using transmission electron microscopy we found that rat hippocampal neurons exposed to 0.5 mM N-methyl-D-aspartate for 10 min produced more cytoplasmic vacuolization than in control neurons. Chronic treatment with mood stabilizers, lithium, valproate or carbamazepine for 7 days at therapeutically relevant concentrations fully attenuated N-methyl-D-aspartate-mediated cytoplasmic vacuolization. These results suggest that inhibition of neurotoxicity may be involved in the action of mood stabilizers. PMID- 12654345 TI - 5-hydroxytryptamine(2A) receptors regulate sympathetic nerves constricting the cutaneous vascular bed in rabbits and rats. AB - Hyperthermia induced by 3,4-methylenedioxymethamphetamine (MDMA) is partially due to sympathetically-mediated cutaneous vasoconstriction that impairs normal heat dissipation. MDMA acts by releasing monoamines, including 5-hydroxytryptamine (5 HT), but receptor mechanisms underlying MDMA-elicited hyperthermia and cutaneous vasoconstriction are not known. The specific 5-HT2A agonist (+/-)-1-(2,5 dimethoxy-4-iodophenyl)-2-aminopropane (DOI) is a potent hallucinogen that also causes marked hyperthermia, suggesting the possibility that DOI, via stimulation of 5-HT2A receptors, might also cause sympathetically mediated cutaneous vasoconstriction. We tested this hypothesis in conscious unrestrained rabbits and rats. Blood flow was assessed by chronically implanted Doppler ultrasonic flow probes. Body temperature was measured by i.p. telemetric probes. We compared effects of DOI on cutaneous blood flow (ear pinna in rabbits, tail in rats) with effects on mesenteric blood flow and arterial pressure.Hyperthermia induced by DOI (5-100 microgram/kg i.v. in rabbits and 100 microgram/kg s.c. in rats) was preceded and accompanied by markedly reduced blood flow to the cutaneous bed, with no change in flow to the mesenteric bed. In rabbits, DOI (5 microgram/kg i.v.) did not affect arterial pressure or heart rate. DOI (100 microgram/kg i.v.) caused a moderate rise in arterial pressure. In rabbits, the 5-HT2A receptor antagonists ketanserin (0.3 mg/kg i.v.) and AC90179 (0.5 mg/kg i.v.) reversed the ear pinna vasoconstriction induced by DOI (5 microgram/kg i.v.). In rats, ketanserin (3 mg/kg s.c.) reversed tail vasoconstriction and hyperthermia induced by DOI (100 microgram/kg s.c.). In rabbits, the cutaneous vasoconstricting effect of DOI (5 microgram/kg i.v.) was substantially abolished in the ipsilateral ear pinna after interruption of preganglionic sympathetic nerve activity by unilateral section of the cervical sympathetic trunk. Thus hyperthermia evoked by direct stimulation of 5-HT2A receptors is associated with marked sympathetically mediated vasoconstriction, selective for the cutaneous bed. Impairment of the ability to dissipate heat following drug-induced stimulation of 5-HT2A receptors is likely to contribute to hyperthermia induced by MDMA and by hallucinogenic drugs such as LSD. PMID- 12654348 TI - Long-term changes in hippocampal n-methyl-D-aspartate receptor subunits following unilateral vestibular damage in rat. AB - Previous studies have indicated that damage to the peripheral vestibular system results in dysfunction of hippocampal place cells and an impairment of spatial learning and memory. The aim of this study was to determine whether lesions of one vestibular labyrinth (unilateral vestibular deafferentation, UVD) result in changes in the expression of the NR1 and NR2A subunits of the N-methyl-D aspartate (NMDA) receptor, and the GluR2 subunit of the alpha-amino-3-hydroxy-5 methyl-4-isoxazole propionate receptor, in subregions of the rat hippocampus (CA1, CA2/3 and the dentate gyrus) at 10 h or 2 weeks following UVD. Compared with sham surgery controls and anaesthetic controls, the expression of the NR1 subunit was significantly reduced in the ipsilateral CA2/3 region at 2 weeks post UVD. The expression of the NR2A subunit was also significantly reduced in the ipsilateral CA2/3 and, to a smaller extent, in the contralateral CA2/3 region, at 2 weeks post-UVD. The only other change in NR2A expression was an increase in the ipsilateral CA1 at 10 h post-UVD. No other changes in NR1, NR2A or GluR2 expression were observed in any hippocampal subregion, at any time point, or in cortical tissue at any time point. These results suggest that UVD may result in long-term changes in NMDA receptor subunit expression in the rat hippocampus. PMID- 12654349 TI - Estrous odors and sexually conditioned neutral odors activate separate neural pathways in the male rat. AB - Olfactory stimuli play important roles in sexual behavior. Previous studies have demonstrated that both estrous odors and initially neutral odors paired with copulation influence the sexual behavior of male rats. The present study examines the pattern of neural activation as revealed by Fos immunoreactivity (Fos-IR) following exposure to bedding scented with either a neutral odor (almond) paired previously with copulation, estrous odors or no odor. Following exposure to estrous odors Fos-IR increased in the accessory olfactory bulb, medial amygdala, medial bed nucleus of the stria terminalis, medial preoptic area, ventromedial hypothalamus, ventral tegmental area, and both the nucleus accumbens core and shell. Conversely, following exposure to the sexually conditioned odor Fos-IR increased in the piriform cortex, basolateral amygdala, nucleus accumbens core, and the anterior portion of the lateral hypothalamic area. In addition, following exposure to almond odor Fos-IR increased in the main olfactory bulb independent of its pairing with copulation. These patterns of Fos-IR following exposure to estrous or sexually conditioned odors were not influenced by either the addition or omission of the other type of odor. These findings demonstrate that estrous and sexually conditioned odors are processed by distinct neural pathways and converge in the nucleus accumbens core, suggesting that this structure has a unique role in processing sexual stimuli of both pheromonal and olfactory natures. PMID- 12654350 TI - Cutaneous sympathetic motor rhythms in the decerebrate rat. AB - Investigation of rhythmic discharges may provide insights into integrative mechanisms underlying nervous system control of effectors. We have previously shown that, in CNS-intact, anesthetized rats, cutaneous sympathetic vasoconstrictor neurones innervating thermoregulatory circulations exhibit a robust rhythmicity in the 0.4-1.2-Hz frequency range (T-rhythm). Here we examined whether the neural circuitry required to generate this rhythm remained intact in decerebrate (at collicular level), paralyzed and artificially ventilated preparations with cervical vagotomy, ligation of common carotid arteries and pneumothorax. Population sympathetic activity was recorded from the ventral collector nerve (VCN) of the tail in nine animals, while monitoring central respiratory drive. We found that rhythmic activity remained a robust feature and that activity behaved in a comparable manner to that previously described in the intact anesthetized preparation. Manifest as peaks in the autospectra, the dominant rhythm was either at the frequency of (f) lung inflation cycle (fLIC), central respiratory drive (fCRD) or in the 'free-run' T-rhythm frequency range. Through manipulation of fLIC we could alter the dominant rhythm of discharges. We show a significant relationship between fLIC and the likelihood of the dominant rhythm in VCN discharges being at fLIC or at a frequency that was neither fLIC nor fCRD. At fLIC of 1 Hz: in seven of nine animals the VCN dominant rhythm was 1 Hz, zero of nine displayed a dominant T-rhythm; at fLIC of 2 Hz: two of nine had a dominant VCN rhythm at 2 Hz and five of nine a T-rhythm. Furthermore, CRD was never observed to entrain to fLIC. These experiments demonstrate that the network underlying the generation of the T-rhythm is located below the collicular level of the neuraxis and that in this preparation LIC-related modulation of discharges may be mediated by spinal (sympathetic) afferents. PMID- 12654351 TI - Distribution of glutamate receptor subunits in experimentally induced cortical malformations. AB - Electrophysiological studies in humans and animal models have revealed an intrinsic epileptogenicity of cortical dysplasias which are a frequent cause of drug-resistant epilepsy. An imbalance of inhibition and excitation has been causative related. Receptor-binding studies in rodents demonstrated reduced binding to GABA and increased binding to glutamate receptors within cortical dysplasias and increments of AMPA- and kainate-receptor binding in its surround. Immunohistochemically a differential downregulation of GABA(A) receptor subunits could be demonstrated in widespread areas within and around dysplasias. As receptor binding critically depends on receptor subunit composition the observed changes in binding properties might be related to this. Here, we immunohistochemically analyzed the regional expression of four NMDA receptor subunits and two major AMPA- and kainate-receptor complexes in adult rats after neonatal freeze lesions. These lesions are characterized by a three- to four layered cortex and a microsulcus which mimic human polymicrogyria. Using antibodies against NR1, NR2A, NR2B, NR2D, GluR2,3, and GluR5,6,7 receptor subunits we demonstrated a pronounced disturbance of cortical immunostaining pattern in the cortical malformation. These changes reflected the structural disorganization of the microgyrus with some distortion of the apical dendrites of paramicrogyral pyramidal cells, a decrease and disorganization of cells at the bottom of the microsulcus, and an inflection of apical dendrites toward the microsulcus. The neuronal staining pattern of large pyramidal cells in the neighborhood of the dysplasia did not differ for any subunit investigated. No remote or widespread changes of glutamate-receptor subunit distribution could be detected. The lack of gross and/or widespread alterations of glutamate-receptor subunit distribution in the surround of focal cortical dysplasia suggests the presence of other or additional mechanisms underlying the increased excitatory neurotransmitter binding and excitability in cortical malformations. PMID- 12654347 TI - Activity-dependent regulation of the subcellular localization of neuronal calcium sensor-1 in the avian cochlear nucleus. AB - Neurons in the avian cochlear nucleus, nucleus magnocellularis (NM), are highly sensitive to manipulations of afferent input, and removal of afferent activity through cochlear ablation results in the death of approximately 20-40% of ipsilateral NM neurons. The intracellular cascades that determine whether an individual NM neuron will die or survive are not fully understood. One early event observed in NM following deafferentation is a rapid rise in intracellular calcium concentration. In most cellular systems, the activity of calcium-binding proteins is believed to accommodate calcium influx. The calcium-binding protein, neuronal calcium sensor-1 (NCS-1), is an intracellular neuronal calcium sensor belonging to the EF-hand superfamily. NCS-1 has been implicated in calcium dependent regulation of signaling cascades. To evaluate NCS-1 action in NM neurons, the localization of NCS-1 protein was examined. Double-label immunofluorescence experiments revealed that NCS-1 expression is evident in both the presynaptic nerve terminal and postsynaptic NM neuron. The postsynaptic expression of NCS-1 typically appears to be closely associated with the cell membrane. This close proximity of NCS-1 to the postsynaptic membrane could allow NCS-1 to function as a modulator of postsynaptic signaling events. Following deafferentation, NM neurons were more likely to show diffuse cytoplasmic NCS-1 labeling. This increase in the number of cells showing diffuse cytoplasmic labeling was observed 12 and 24 h following cochlea ablation, but was not observed 4 days following surgery. This activity-dependent regulation of NCS-1 subcellular localization suggests it may be associated with, or influenced by, processes important for the survival of NM neurons. PMID- 12654352 TI - Auditory thalamic nuclei projections to the temporal cortex in the rat. AB - Thalamocortical projections from the auditory thalamic nuclei were examined systematically in the rat, including those from the dorsal division (MGD) of the medial geniculate body (MG), which were less clearly determined in previous studies. Injections of biocytin confined in each thalamic nucleus revealed characteristic features of projections in terms of cortical areas and layers of termination. In contrast to exclusively selective projections to cortical area Te1 from the ventral division (MGV) of the MG, diffuse and selective terminations were observed in the projections from the dorsal (MGD) and medial divisions (MGM) of the MG and the suprageniculate nucleus (SG). Diffuse termination was continuous in layer I or VI of the temporal cortex, while selective termination was in layers III and IV of discrete cortical areas. In addition to diffuse termination in the upper half of layer I of cortical areas Te1, Te2d and Te3v, the MGD and SG projections formed plexuses of axons selectively in lower layer III and layer IV of Te2d and Te3v. The SG projections targeted further the dorsal bank of the perirhinal cortex (PRh), while the MGD projections targeted in part the ventral fringe of Te1. The MGM projections terminated diffusely in layer VI of Te1 and Te3v, and selectively in lower layer III and layer IV of the rostral part of Te3v. Diffuse projections to layers I and VI from the SG and MGM extended in cortical regions over the dorsal fringe of Te1. Selective dense projections to middle cortical layers of Te2d and Te3v (especially its rostral part) indicate the existence of auditory areas, which could be involved in cross-modal interaction with visual and somatosensory system, respectively. Diffuse projections are supposed to bind information processings in these areas and the primary auditory area (Te1). PMID- 12654353 TI - Enhancement of 5-hydroxytryptamine-induced head-twitch response after olfactory bulbectomy. AB - 5-Hydroxytryptamine(2A) receptor agonists evoke the head-twitch response in mice. The head-twitch response in olfactory bulbectomized mice elicited by the administration of 5-hydroxytryptamine (40 microgram/mouse, i.c.v.) was increased about threefold as compared with controls on the 14th day after the operation. The injection of ketanserin (1 mg/kg, i.p.), a 5-hydroxytryptamine(2A) receptor antagonist, inhibited this enhancement of 5-hydroxytryptamine-induced head-twitch response after olfactory bulbectomized. On the 14th day, the number of head twitch response induced by 5-hydroxytryptophan (40, 80 and 160 mg/kg, i.p.), a precursor of 5-hydroxytryptamine, did not differ between olfactory bulbectomized and control mice. Monoamine oxidase-B activity in the forebrain of olfactory bulbectomized mice was higher than that in controls while monoamine oxidase-A activities were unchanged. The 5-hydroxytryptamine uptake into synaptosomes in the forebrain homogenates of olfactory bulbectomized mice was lower than that in controls. These findings indicate that olfactory bulbectomized causes the enhancement of head-twitch response by a supersensitivity of 5 hydroxytryptamine(2A) receptors in cerebral cortex derived from degeneration of neurons projecting from the olfactory bulb. PMID- 12654354 TI - Contribution of hippocampal place cell activity to learning and formation of goal directed navigation in rats. AB - Although extensive behavioral studies have demonstrated that hippocampal lesions impair navigation toward specific places, the role of hippocampal neuronal activity in the development of efficient navigation during place learning remains unknown. The aim of the present study was to investigate how hippocampal neuronal activity changes as rats learn to navigate efficiently to acquire rewards in an open field. Rats were pre-trained in a random reward task where intracranial self stimulation rewards were provided at random locations. Then, the rats were trained in a novel place task where they were rewarded at two specific locations as they repeatedly shuttled between them. Hippocampal neuronal activity was recorded during the course of learning of the place task. The rats learned reward sites within several sessions, and gradually developed efficient navigation strategies throughout the learning sessions. Some hippocampal neurons gradually changed spatial firing as the learning proceeded, and discharged robustly near the reward sites when efficient navigation was established. Over the learning sessions, the neuronal activity was highly correlated to formation of efficient shuttling trajectories between the reward sites. At the end of the experiment, spatial firing patterns of the hippocampal neurons were re-examined in the random reward task. The specific spatial firing patterns of the neurons were preserved if the rats navigated, as if they expected to find rewards at the previously valid locations. However, those specific spatial firing patterns were not observed in rats pursuing random trajectories. These results suggest that hippocampal neurons have a crucial role in formation of an efficient navigation. PMID- 12654355 TI - Prolonged exercise induces angiogenesis and increases cerebral blood volume in primary motor cortex of the rat. AB - Plastic changes in motor cortex capillary structure and function were examined in three separate experiments in adult rats following prolonged exercise. The first two experiments employed T-two-star (T(2)*)-weighted and flow-alternating inversion recovery (FAIR) functional magnetic resonance imaging to assess chronic changes in blood volume and flow as a result of exercise. The third experiment used an antibody against the CD61 integrin expressed on developing capillaries to determine if motor cortex capillaries undergo structural modifications. In experiment 1, T(2)*-weighted images of forelimb regions of motor cortex were obtained following 30 days of either repetitive activity on a running wheel or relative inactivity. The proton signal intensity was markedly reduced in the motor cortex of exercised animals compared with that of controls. This reduction was not attributable to alterations of vascular iron levels. These results are therefore most consistent with increased capillary perfusion or blood volume of forelimb regions of motor cortex. FAIR images acquired during experiment 2 under normocapnic and hypercapnic conditions indicated that resting cerebral blood flow was not altered under normal conditions but was elevated in response to high levels of CO(2), suggesting that prolonged exercise increases the size of a capillary reserve. Finally, the immunohistological data indicated that exercise induces robust growth of capillaries (angiogenesis) within 30 days from the onset of the exercise regimen. Analysis of other regions failed to find any changes in perfusion or capillary structure suggesting that this motor activity-induced plasticity may be specific to motor cortex.These data indicate that capillary growth occurs in motor areas of the cerebral cortex as a robust adaptation to prolonged motor activity. In addition to capillary growth, the vascular system also experiences heightened flow under conditions of activation. These changes are chronic and observable even in the anesthetized animal and are measurable using noninvasive techniques. PMID- 12654356 TI - Botulinum A toxin injection for restrictive myopathy of thyroid-related orbitopathy: effects on intraocular pressure. AB - PURPOSE: To study the effect of extraocular muscle injections of botulinum A toxin on intraocular pressure in patients with thyroid-related orbitopathy. DESIGN: Retrospective observational case series. METHODS: The medical records of eight consecutive patients with restrictive myopathy secondary to thyroid related orbitopathy (TRO) who underwent botulinum A toxin injection from December 1997 to December 1998 were reviewed and analyzed retrospectively. All patients were seen at the University of California, San Diego (UCSD) Thyroid Eye Center, a university-based tertiary referral center. The main outcome measure was intraocular pressure (IOP) readings taken before and after injection in both primary gaze and upgaze (involving one eye in seven of the patients and both eyes in one patient). Intraocular pressure readings were measured by an unmasked physician using a Goldmann applanation tonometer. RESULTS: A statistically significant decrease in IOP in upgaze was noted 2 to 6 weeks following botulinum A toxin injection and in both fields of gaze (primary and upgaze) after 2 to 4 months. The mean IOP before injection was 21.4 +/- 3.0 mm Hg in primary gaze and 29.9 +/- 9.7 mm Hg in upgaze. The mean IOP, following injection at 2 to 6 weeks, was 19.2 +/- 4.2 mm Hg (P <.095) in primary gaze and 25.1 +/- 5.9 mm Hg (P <.023) in upgaze. At 2 to 4 months following injection, the mean IOP was 19.3 +/- 3.9 mm Hg (P <.044) in primary gaze and 27.7 +/- 8.5 mm Hg (P <.024) in upgaze. Six patients indicated improved ocular deviation, which was associated with a lowering of IOP. Two patients indicated no change in IOP or strabismic deviation following botulinum A toxin injection. CONCLUSIONS: Botulinum A toxin injections cause a secondary effect to lower IOP in patients with restrictive strabismus associated with thyroid-related orbitopathy. PMID- 12654357 TI - Aging changes in apparent accommodation in eyes with a monofocal intraocular lens. AB - PURPOSE: To examine the aging changes in the amplitude of apparent accommodation in eyes with a monofocal intraocular lens (IOL). DESIGN: Prospective comparative observational study. METHODS: Two hundred eyes of 200 patients who were scheduled for phacoemulsification and monofocal IOL implantation were studied. Forty patients in each of five age groups-younger than 40 years of age or in their 50s, 60s, 70s, and 80s-were prospectively recruited. Using an accommodopolyrecorder, the amplitude of apparent accommodation of these patients was measured approximately 1 month after surgery. Visual acuities from far to near distances after best-corrected distance were examined using an all-distance vision tester, and the region of accommodation at which each patient achieved a visual acuity of 20/29 or 20/40 was determined and converted to the diopteric range. Contrast sensitivity was also examined using a vision contrast test system. RESULTS: The amount of apparent accommodation as measured with the accommodopolyrecorder was decreased significantly in proportion to patient age (P <.0001). Furthermore, significant negative correlation was found between the amount of apparent accommodation and actual age of each patient (r = -.491). The diopteric range of accommodation at which the patients achieved 20/29 or 20/40 also reduced in proportion to the age. Although no significant differences were found in the mean far visual acuities between age groups, the intermediate and near visual acuities worsened significantly with age. The contrast sensitivity was also reduced in proportion to the age. CONCLUSIONS: The amplitude of apparent accommodation in eyes with a monofocal IOL decreases significantly in proportion to age, resulting in worsening of best-corrected intermediate and near visual acuities. PMID- 12654358 TI - Anemia and papilledema. AB - PURPOSE: To elucidate the relationship between anemia and raised intracranial pressure (ICP). DESIGN: Interventional case series. METHODS: Retrospective case series and review of the literature. Only patients with documented papilledema, neuroimaging ruling out a space-occupying lesion, and anemia were included. RESULTS: Five women with confirmed idiopathic intracranial hypertension (IIH) (normal brain magnetic resonance imaging, normal cerebrospinal fluid, elevated intracranial pressure), and one man with presumed IIH (normal head computed tomography [CT], no lumbar puncture) were evaluated. All had bilateral papilledema associated with peripapillary hemorrhages. Two had retinal cotton wool spots (CWS), and two had preretinal hemorrhages. All had severe iron deficiency anemia, which was discovered at the time of their ocular complaints in five of them. Their symptoms and signs improved dramatically after treatment of the anemia. We found 30 well-documented cases in the English and French literature. Among those, 13 were excluded from our analyses (11 had confounding disorders, and two had cerebral venous thrombosis). In the remaining 17 cases, isolated raised ICP associated with anemia was the most likely diagnosis, although in none of these cases was cerebral venous thrombosis excluded. CONCLUSIONS: Anemia may play a role in the occurrence of raised ICP and papilledema. Although only a few cases in the literature support this association, it may be more common than previously thought. Because most patients are not known to be anemic when papilledema is discovered, we suggest that a complete blood count be obtained in patients with IIH, especially in the absence of known associated factors such as obesity or medications or when treatment aimed at lowering ICP fails to improve the patient's symptoms. The underlying mechanisms remain unknown, but cerebral venous thrombosis should be carefully excluded. PMID- 12654359 TI - Azithromycin vs doxycycline in the treatment of inclusion conjunctivitis. AB - PURPOSE: The aim of the study was to compare the efficacy and safety of azithromycin and doxycycline in the treatment of chlamydial conjunctivitis in adults. DESIGN: An open, randomized clinical trial. METHODS: Seventy-eight adult patients with incluson conjunctivitis were enrolled in this multicenter clinical study. Patients with chlamydial conjunctivitis as indicated by a positive direct fluorescent antibody (DFA) test or cell culture were randomized to receive a single 1-g dose of azithromycin or doxycycline, 100 mg twice daily for 10 days. A conjuctival swab for cell culture was obtained from all patients immediately before the treatment for subsequent confirmation of the presence of chlamydial infection in the central laboratory. Control examinations were performed 10 to 12 days and 4 to 6 weeks after the treatment initiation. Clinical and bacteriological responses to the treatment were evaluated at the last visit. The occurrence and frequency of adverse events were analyzed as well. RESULTS: Of 78 patients enrolled, 51 completed the study and were evaluated for efficacy. The main reasons for withdrawal were lack of confirmation of the presence of chlamydial infection by the central laboratory and failure to attend the follow up visit. Eradication of C. trachomatis was achieved in 23 of 25 (92%) patients treated with azithromycin and in 25 of 26 (96%) patients treated with doxycycline. Clinical cure was observed in 15 (60%) and 18 (69%) patients treated with azithromycin and doxycycline, respectively. Both drugs were equally well tolerated. CONCLUSIONS: A single 1-g azithromycin therapy was as effective as standard 10-day treatment with doxycycline (100 mg twice daily) in the treatment of adult inclusion conjunctivitis. PMID- 12654360 TI - Preliminary findings in corneal allograft rejection in patients with keratoconus. AB - PURPOSE: Classically, corneal allograft rejection is thought to be a T(H)1 mediated phenomenon. However, T(H)2-mediated allograft rejection has been reported in other transplanted organ systems, including the heart and kidney. We previously reported a form of T(H)2-mediated corneal allograft rejection in a murine model with a T(H)2 immune bias. In this study we sought to determine if there was any evidence for this form of corneal allograft rejection in humans. DESIGN: Experimental study with an interventional case series. METHODS: The clinical records of all keratoconus patients undergoing penetrating keratoplasty at the University of Texas, Southwestern Medical Center from 1994 to 1999 were reviewed. Careful attention was paid to a clinical history of atopy. Atopic patients were selected, because these patients have been shown to have a "T(H)2 immune bias." The corneal graft rejection rate in these patients and the number of repeat corneal transplants performed was determined. The experimental group consisted of patients with a clinical history of atopy and keratoconus who had at least one repeat penetrating keratoplasty for an immunologically rejected corneal transplant. Any patient with evidence of primary allograft failure was excluded from this study. Tissue specimens from these patients were embedded in paraffin, serially sectioned, stained with Giemsa stains, and examined histologically. The control group consisted of patients without a clinical history of allergy (and therefore no T(H)2 immune bias) who underwent corneal transplantation for Fuch corneal endothelial dystrophy, or aphakic/pseudophakic bullous keratopathy. Failed grafts from these control patients were also paraffin embedded, serially sectioned, stained, and examined histologically. The human experimental and control corneal specimens were compared with data obtained in a murine model of T(H)2-mediated corneal allograft rejection. Briefly, full-thickness penetrating C57BL/6ByJ corneal allografts were transplanted onto Balb/cByJ and Balb/c-IFN gamma(tm1Ts) (Balb/c-IFN-gamma knockout) mice. Additionally, full-thickness Balb/cByJ corneal allografts were transplanted onto C57BL/6ByJ and C57BL/6ByJ-IFN gamma(tm1Ts) mice. Corneal allograft rejection rates and mean rejection times were calculated and compared between wild-type and interferon gamma (IFN-gamma) knockout hosts. The rejected allografts were examined histologically by the same methods used in the human tissue. RESULTS: There were 84 penetrating keratoplasties performed from 1994 to 1999 for keratoconus. Seven of these 84 patients rejected their corneal grafts. Of the 7 patients who rejected their corneal allografts, 4 had repeat penetrating keratoplasty. Of these 4 repeat corneal allografts, 3 showed eosinophilia when compared with rejected grafts in control patients. Atopic keratoconus patients had a mixed inflammatory cellular infiltrate in the rejected corneal tissue specimen with a significantly greater density of eosinophils (P =.001) compared with patients who did not have a pre existing T(H)2 bias. The inflammatory infiltrate in these patients without a T(H)2 immune bias was mononuclear. In the murine model, corneal allograft rejection did occur in the absence of IFN-gamma, a critical T(H)1 cytokine in both fully allogeneic donor-host combinations. Histologically, rejection in these ("T(H)2 mice") was characterized by a predominant eosinophilic infiltrate in the rejected graft bed when compared with wild-type animals ("T(H)1 mice") that had a predominantly mononuclear infiltrate in the rejected corneal graft bed. CONCLUSIONS: Preliminary findings show that corneal allograft rejection in patients with a pre-existing T(H)2 phenotype is similar to what is seen in the murine model of T(H)2-mediated corneal allograft rejection. Based on this small sample, it appears that eosinophils may play a role in corneal allograft rejection in this group of patients. However, further study is necessary to determine the importance of these cells in allograft rejection. PMID- 12654362 TI - The magnitude and cost of global blindness: an increasing problem that can be alleviated. AB - PURPOSE: To identify the potential effect on global economic productivity of successful interventions, that are planned as part of the "VISION 2020-right to sight" initiative. The initiative aims to eliminate avoidable blindness. DESIGN: This study used economic and epidemiologic modeling. METHODS: Existing data and assumptions about blindness prevalence, national populations, gross domestic product (GDP) per capita, labor force participation, and unemployment rates were used to project the economic productivity loss associated with unaccommodated blindness. RESULTS: Without extra interventions, the global number of blind individuals would increase from 44 million in the year 2000 to 76 million in 2020. A successful VISION 2020 initiative would result in only 24 million blind in 2020 and lead to 429 million blind person-years avoided. A conservative estimate of the economic gain is $102 billion. CONCLUSIONS: The VISION 2020 initiative has the potential to increase global economic productivity. PMID- 12654361 TI - Clinicopathologic correlation and genetic analysis in a case of posterior polymorphous corneal dystrophy. AB - PURPOSE: To evaluate the clinical history, histopathology, and genetics of posterior polymorphous corneal dystrophy (PPMD) in a woman with a prominent retrocorneal membrane. DESIGN: Observational case report and genetic analysis of her family, UM:139. METHODS: Records were reviewed from a case and associated family members. The diagnosis of PPMD was based on clinical examination, immunohistochemical staining, electron microscopy, and screening of genetic markers from regions previously reported to be associated with PPMD. RESULTS: Over 17 years, the proband with PPMD had 25 ocular procedures performed for glaucoma, cataract, cornea, retina, and postoperative problems. A prominent retrocorneal membrane grew onto the crystalline lens and intraocular lens (IOL). Histopathology revealed stratified epithelial-like cells on iris from an iridectomy and stratified corneal endothelium on a corneal button. Electron microscopy on the cornea revealed microvilli, tonofilaments, and desmosomes consistent with endothelial transformation, which was confirmed by positive anticytokeratin (CK) AE1/AE3 and CAM 5.2 immunoreactivity. Negative immunoreactivity in epithelium and positive in endothelium with anti-CK 7 supported the diagnosis of PPMD rather than epithelial downgrowth. Multiple relatives were affected with PPMD with apparent autosomal dominant inheritance, but surprisingly, the PPMD, congenital hereditary endothelial dystrophy 1 (CHED1) and CHED2 loci on chromosome 20 and the collagen, type VIII, alpha-2 (COL8A2) gene were excluded by linkage and haplotype analyses. CONCLUSIONS: We are unaware of previous PPMD reports describing the unusual feature of a retrocorneal membrane extending onto the crystalline lens and IOL. In addition, this family suggests another PPMD locus. PMID- 12654363 TI - Reduced oscillatory potentials of the full-field electroretinogram of eyes with aphakic or pseudophakic cystoid macular edema. AB - PURPOSE: To report reduced oscillatory potentials (OPs) in the full-field electroretinogram (ERG) of eyes with aphakic or pseudophakic cystoid macular edema (CME). DESIGN: Observational case series. METHODS: Bright white flash full field ERGs were recorded from 19 eyes of 19 patients (ages, 53-84 years) with aphakic or pseudophakic CME. Seven of the cases had uncomplicated phacoemulsification (PE) with implantation of a posterior chamber intraocular lens (PC-IOL) and 6 had posterior capsular rupture during phacoemulsification and aspiration with implantation of a PC-IOL. Four additional patients had uncomplicated intracapsular cataract extraction (ICCE), and 2 had secondary anterior chamber intraocular lens after ICCE. Their fellow eyes without CME served as controls. Cystoid macular edema was diagnosed as clinically significant by slit-lamp examination and by fluorescein angiography. The grading for severity of CME was based on the fluorescein angiography. The visual acuity ranged from 20/200 to 20/16. RESULTS: There were no significant differences in the mean amplitudes and implicit times of the a-waves and b-waves between the affected eyes and the fellow eyes. The mean summed amplitude of the OPs was significantly reduced in the affected eyes (P =.0003, Wilcoxon signed rank test). This decrease was strongly correlated with visual acuity (r =.779, P <.0001). The mean implicit time of the first OP was significantly delayed in the affected eyes (P =.0089, Wilcoxon signed rank test). CONCLUSIONS: Because the peripheral retina contributes more significantly to full-field ERGs than the macula, the reduced OP amplitudes of the full-field ERGs in eyes with aphakic or pseudophakic CME suggest a functional impairment not only in the macula but also throughout the retina. PMID- 12654364 TI - Retinal arterial wall plaques in Susac syndrome. AB - PURPOSE: To demonstrate retinal arterial wall plaques (RAWPs) in patients with Susac syndrome, a disorder that consists of the triad of branch retinal artery occlusion, encephalopathy, and hearing loss. The clinician may misinterpret these RAWPs as emboli. DESIGN: Observational case series. METHODS: Four patients (one man and three women, aged 21-38 years) were examined and followed. RESULTS: Four patients with Susac syndrome had RAWPs in association with branch retinal artery occlusions. They were present in six of eight eyes. The plaques were yellow to yellow-white and located usually away from retinal bifurcations. Retinal arterial wall plaques occur at the midarteriolar segments, whereas Hollenhorst plaques occur at vascular bifurcations. Four of six eyes had resolution of the plaques. CONCLUSIONS: Retinal arterial wall plaques may occur with branch retinal artery occlusions in Susac syndrome. This finding should help the clinician who is struggling to make a diagnosis in a patient with an enigmatic encephalopathy and/or hearing loss. These plaques may resolve over time. PMID- 12654365 TI - Macular hole formation following ruptured retinal arterial macroaneurysm. AB - PURPOSE: To report four cases of macular hole formation following ruptured retinal arterial macroaneurysm. DESIGN: Case-control study. METHODS: Review of clinical records of 75 consecutive eyes (73 patients) with macroaneurysm. Macroaneurysms were confirmed using either fluorescein angiography or indocyanine green angiography or both that indicated a telangiectatic retinal vascular change. Clinical data of patients with macroaneurysm with and without macular hole (MH) formation were compared retrospectively. RESULTS: Of 75 eyes, four (5.3%; four patients) with macroaneurysm had a macular hole. In one of the four eyes, a macular hole was detected 2 days after the rupture. The distance from the macroaneurysm to the center of the fovea was significantly closer in the four eyes with a macular hole (MH group, 1.1 +/- 0.4 (mean +/- standard deviation [SD]) disk diameter) than in the 71 eyes without a macular hole (non-MH group, 1.9 +/- 0.9 disk diameter) (P =.0196). The incidence of subretinal hemorrhage in the macula was significantly greater in the MH group (100%) than in the non-MH group (35.2%) (P =.0195). CONCLUSIONS: Macular hole formation associated with ruptured macroaneurysm is not as uncommon as previously thought. We suggest that there might be early onset of macular hole formation following ruptured macroaneurysm. Macroaneurysms with early-onset macular hole are present near the center of the macula and are accompanied by subretinal hemorrhage in the macula. PMID- 12654366 TI - Measured visual acuity of fellow eyes as a prognostic factor in macular hole surgery. AB - PURPOSE: To investigate the effect of the vision of the fellow eye on the visual acuity of eyes after successful macular hole surgery. DESIGN: Prospective consecutive series. METHODS: A consecutive series of eyes with successful macular hole closure were studied and assigned to one of two groups according to the visual acuity of the fellow eye; a group with visual acuity less than 20/200 and a group with visual acuity of 20/200 or better. Preoperative and postoperative visual acuity in the two groups was measured based on the logarithm of the minimal angle of resolution (LogMAR), and postoperative visual acuity was also determined by using the multiple-letter visual acuity chart, which permitted measurement of visual acuity at an extrafoveal point. RESULTS: Group 1 (<20/200) consisted of 19 eyes and group 2 (>20/200) consisted of 51 eyes. LogMAR visual acuity at 6 months postoperatively was significantly better in group 1 than in group 2 (0.21 vs 0.41, P <.01). The logMAR change 6 months after surgery was significantly greater in group 1 than in group 2 (0.49 vs 0.23, P <.01). There was no significant difference between two groups in logMAR visual acuity at 6 months postoperatively determined with the multiple-letter visual acuity chart (0.20 vs 0.29, P >.05). CONCLUSIONS: Visual recovery after successful macular hole surgery is inversely correlated with vision in the fellow eye. Learning to use eccentric fixation may contribute to visual improvement after macular hole surgery. PMID- 12654367 TI - Characteristics of visual field progression in patients with normal-tension glaucoma with optic disk hemorrhages. AB - PURPOSE: To study the characteristics of visual field progression in patients with normal-tension glaucoma (NTG) with optic disk hemorrhages. DESIGN: Observational study. METHODS: Fifty-eight eyes of 58 untreated patients with NTG who had at least five reliable visual fields of the Humphrey Field Analyzer (central 30-2) within the follow-up period of more than 24 months were enrolled. Of these, 27 eyes had optic disk hemorrhages in their clinical courses. Pointwise linear regression analysis was done using total deviation values of the fields at each of 74 test locations in each patient. Progression was defined as the points of negative slopes with P <.01. A whole visual field was concentrically divided into three clusters: the areas within 10 degrees, 10 to 20 degrees, and 20 to 30 degrees. Percentages of the progressed points of the three clusters and a whole field were compared between the groups with and without optic disk hemorrhages. RESULTS: No significant differences were found in patients' backgrounds, including initial mean deviation values, follow-up periods, and the number of visual field examinations between the 27 patients with optic disk hemorrhages and the 31 without. The group with optic disk hemorrhages showed significantly higher percentages of progressed points within the 10-degree area compared with the group without optic disk hemorrhages (mean +/- SD: 13.1 +/- 13.7%; 5.1 +/- 8.5%, respectively; P =.0086, Student t test), whereas no significant differences were found in the other two clusters or in a whole field. CONCLUSIONS: Patients with NTG with optic disk hemorrhages tend to show visual field progression in areas within 10 degrees. PMID- 12654368 TI - Comparison of optic nerve head measurements obtained by optical coherence tomography and confocal scanning laser ophthalmoscopy. AB - PURPOSE: To evaluate the relationship between optic nerve head (ONH) measurements generated by optical coherence tomography (OCT; versions 2 and 3) and confocal scanning laser ophthalmoscopy (CSLO) and to compare the association between OCT and CSLO ONH measurements with glaucoma disease status, as determined by clinical evaluation and perimetry. DESIGN: Cross-sectional study. METHODS: In a prospective study in the glaucoma service of an academic department of ophthalmology, 159 eyes (97 subjects) and 77 eyes (44 subjects) were recruited in two separate periods. All subjects were scanned with a CSLO device. Subjects tested within the first period of recruitment were scanned with OCT version 2 and in the second period with OCT version 3. The main outcome measure was the correlation between automatic and manually defined OCT ONH measurements and the correlation of CSLO and OCT ONH measurements between devices and with glaucoma disease status. RESULTS: A high correlation was found between ONH measurements obtained by the automatic determination of ONH margin and those obtained by manual tracing of the disk margin (r =.93 to.98). Optical coherence tomography and CSLO ONH measurements were highly correlated. Optical coherence tomography measured mean disk area was significantly larger than that measured by CSLO, as were all other disk size-related parameters. The areas under the receiver operator characteristic (AROC) curves for the associations between CSLO and OCT ONH measurements and clinical diagnosis were found to be similar and in the range of 0.47 to 0.79 for both devices. CONCLUSIONS: Automated OCT ONH measurements correlate highly with those obtained by manual tracing of disk margin. Optical coherence tomography and CSLO ONH analyses are highly correlated and have similar associations with glaucoma disease status. PMID- 12654369 TI - Evaluation of the glaucomatous damage on retinal nerve fiber layer thickness measured by optical coherence tomography. AB - PURPOSE: To evaluate the relationship between visual field and retinal nerve fiber layer (RNFL) thickness measured by optical coherent tomography (OCT) and to assess the diagnostic ability of OCT to distinguish between early glaucomatous or glaucoma-suspect eyes from normal eyes. DESIGN: Retrospective, non-randomized, cross-sectional study. METHODS: A total of 160 eyes of 120 normal Japanese adults, 23 eyes of 16 patients with ocular hypertension, 38 eyes of 35 glaucoma suspect patients, and 237 glaucomatous eyes of 140 glaucoma patients were enrolled in the study. The glaucoma group included 89 early glaucomatous eyes. Thickness of the RNFL around the optic disk was determined with three 3.4-mm diameter circle OCT scans. Average and segmental RNFL thickness values were compared among all groups. The correlation between mean deviation and RNFL thickness in glaucomatous eyes was also analyzed. Receiver operating characteristic (ROC) curve area was calculated to discriminate normal eyes from early glaucomatous or glaucoma-suspect eyes. RESULTS: A significant relationship existed between the mean deviation and RNFL thickness in all parameters excluding the 3-o'clock area. The average RNFL thickness had the strongest correlation in all parameters (r = -0.729, P <.001). Retinal nerve fiber layer thickness at the 7-o'clock inferotemporal segment had the widest areas under the ROC curves in all parameters for early glaucomatous eyes (0.873). CONCLUSIONS: Measurement of RNFL thickness by OCT is useful in detecting early RNFL damage. Furthermore, OCT measurements of RNFL thickness may provide clinically relevant information in monitoring glaucomatous changes. PMID- 12654370 TI - Scanning laser polarimetry with variable corneal compensation and optical coherence tomography in normal and glaucomatous eyes. AB - PURPOSE: To evaluate the relationship between visual function and retinal nerve fiber layer (RNFL) measurements obtained with scanning laser polarimetry with variable corneal compensation (SLP-VCC) and optical coherence tomography (OCT). DESIGN: Cross-sectional analysis of normal and glaucomatous eyes in a tertiary care academic referral practice. METHODS: A commercial GDx nerve fiber analyzer was modified to enable the measurement of corneal polarization axis and magnitude so that compensation for corneal birefringence was eye specific. Complete examination, SLP with fixed corneal compensation (FCC) and variable corneal compensation (VCC), optical coherence tomography (OCT) imaging of the peripapillary RNFL, and automated achromatic perimetry were performed in all subjects. Exclusion criteria were visual acuity less than 20/40, diseases other than glaucoma, and unreliable perimetry. RESULTS: Fifty-nine patients (59 eyes; 29 normal, 30 glaucomatous) were enrolled (mean age, 56.7 +/- 20.3 years, range, 20-91). All eyes with glaucoma had associated visual field loss (average mean defect, -8.4 +/- 5.8 dB). Using SLP-FCC, nine of 12 retardation parameters (75%) were significantly less in glaucomatous eyes. Using SLP-VCC, 11of 12 retardation parameters (92%) were significantly less in glaucomatous eyes. Multiple regression models constructed for each retardation parameter with visual field demonstrated that the following VCC parameters were statistically significant whereas FCC parameters were not: ellipse average (FCC, P =.28, VCC, P =.001), superior average (FCC, P =.38, VCC, P <.001), inferior average (FCC, P =.10, VCC, P =.008), average thickness (FCC, P =.30, VCC, P =.031), and superior integral (FCC, P =.43, VCC, P =.001). Similar results were obtained for multiple regression models constructed with OCT-derived RNFL thickness: ellipse average (FCC, P =.99, VCC, P =.002), superior average (FCC, P =.90, VCC, P <.001), inferior average (FCC, P =.61, VCC, P =.007), and superior integral (FCC, P =.92, VCC, P <.001). CONCLUSIONS: Compared with fixed compensation, mean-based SLP parameters generated with SLP-VCC have greater correlation with visual function and RNFL thickness assessments obtained with OCT. PMID- 12654371 TI - An improved technique of eye drop self-administration for patients with limited vision. AB - PURPOSE: To describe a new, highly accurate, tactile technique to increase accuracy in the self-administration of eye drops. DESIGN: Interventional case series. METHODS: Ten adult patients with loss of fixation (< or = 20/400) in one eye and ten adult patients with loss of fixation in both eyes instilled one drop of artificial tears in each eye using the technique normally employed at home. The time required to instill each drop, the number of drops squeezed from the bottle, and location of the drops' landing points on the face or eye were recorded. The patients were then instructed in a new technique for instillation, guided through the procedure once, and allowed to practice until comfortable with it. All measurements were then repeated. RESULTS: In patients with loss of fixation in one eye, an additional 3.2 +/- 4.1 seconds were required to instill a drop using the new technique (P <.0001, paired t test; range, -7-18 seconds). The mean number of drops dispensed decreased by 0.1 +/- 0.6 drops (P =.60, paired t test; range, -2-1 drop). The accuracy of drop placement increased from 80.0% to 82.5% (P =.32, paired t test). In subjects with loss of fixation in both eyes, using the new technique increased the time needed to instill a drop by 3.8 +/- 3.8 seconds (P <.0001, paired t test; range, -3-13 seconds). The average number of drops dispensed decreased by 0.1 +/- 0.6 drops (P =.25, paired t test; range, 2-1 drop). The accuracy of placement increased from 63.0% to 85.0% (P =.001, paired t test). CONCLUSIONS: This technique of drop instillation may be beneficial for patients with significant visual impairment in both eyes. PMID- 12654372 TI - A pilot study of lens extraction in the management of acute primary angle-closure glaucoma. AB - PURPOSE: To describe a pilot study of lens extraction in the management of acute primary angle-closure glaucoma (APACG) in Chinese patients. DESIGN: Interventional case series. METHODS: A prospective case series of patients presented with APACG at a tertiary eye hospital in Xiamen, China. All the patients had intensive medical treatment followed by immediate phacoemulsification and posterior chamber intraocular lens implantation under topical anesthesia. RESULTS: There were 18 patients who were seen with APACG who had phacoemulsification and posterior chamber intraocular lens implantation under topical anesthesia. There was a 9.0-mm Hg drop in mean preoperative and first day postoperative intraocular pressure (22.8 vs 13.8 mm Hg, P =.012) and a 10.2-mm Hg drop in mean preoperative and seventh-day postoperative intraocular pressure (22.8 vs 13.2 mm Hg, P =.002). CONCLUSIONS: This pilot study indicates that phacoemulsification and posterior chamber intraocular lens implantation may be useful in the treatment of primary acute angle-closure glaucoma in Chinese patients. Further study with longer follow-up is needed. PMID- 12654373 TI - Accuracy of intraocular pressure by Tono-Pen XL over amniotic membrane patching in rabbits. AB - PURPOSE: To report an accuracy of intraocular pressure (IOP) by Tono-Pen XL over amniotic membrane patching in rabbits. DESIGN: Experimental study. METHODS: Seven male albino rabbits were used in this study. After general anesthesia, IOP was measured in both eyes using the Tono-Pen without amniotic membrane. A single layer amniotic membrane was then immediately placed over the rabbit eye, and the IOP measurement was repeated. RESULTS: The average IOP without amniotic membrane was 5.8 +/- 0.6 mm Hg (mean +/- SD). The average IOP through amniotic membrane was 6.0 +/- 0.4 mm Hg (mean +/- SD). There was no statistical difference in IOP measured on corneas with or without the amniotic membrane (P =.34). CONCLUSIONS: The accuracy of IOP measurement by Tono-Pen XL over a single layer of amniotic membrane patching was demonstrated using rabbit eyes. Further studies in human eyes may verify our findings in actual clinical settings. PMID- 12654374 TI - Optical coherence tomographic evaluation of a surgically treated traumatic macular hole secondary to Nd:YAG laser injury. AB - PURPOSE: To report the importance of optical coherence tomographic (OCT) evaluation of a macular hole caused by neodymium:yttrium-aluminum-garnet (Nd:YAG) laser injury. DESIGN: Observational case report. METHODS: A 33-year-old man presented with an accidental Nd:YAG laser-induced macular hole in his right eye. After observing the course of the macular hole by OCT, the macular hole was treated by standard pars plana vitrectomy. The appearance of the macular hole was also followed up postoperatively. RESULTS: Seven days after the injury, OCT showed a full-thickness macular hole without a fluid-cuff or swelling of the surrounding retina. Twenty-one days after the injury, however, OCT disclosed that the retina around the macular hole was swollen. After vitrectomy, OCT showed that the macular hole was closed, and the visual acuity improved from 20/30 to 20/20. CONCLUSIONS: Surgical treatment can improve the visual function of eyes with a Nd:YAG laser-induced macular hole. Optical coherence tomography is valuable in providing precise information about the condition of the macula. PMID- 12654375 TI - Lipemia retinalis associated with branch retinal vein occlusion. AB - PURPOSE: To report a patient with a branch retinal vein occlusion associated with lipemia retinalis. DESIGN: Observational case report. METHODS: A 58-year-old woman presented with decreased vision and was found to have a branch retinal vein occlusion with massive lipid exudation in the setting of lipemia retinalis. Laboratory testing demonstrated an abnormal lipid profile with a markedly elevated triglyceride level. The setting was a retina service in a major referral center. RESULTS: Management of the hypertriglyceridemia with medication, exercise, and dietary modification resolved the lipemia retinalis and was associated with improvement in visual function. CONCLUSIONS: Although usually not visually significant, lipemia retinalis may be associated with vascular pathology, such as a branch retinal vein occlusion with marked exudative response and decreased visual acuity. Because of potential systemic and ocular complications of lipemia retinalis, these patients should be referred for management of their lipid disorder. PMID- 12654376 TI - Antiphospholipid antibody syndrome in a six-year-old female patient. AB - PURPOSE: To report the first instance of primary antiphospholipid antibody (APA) syndrome in an otherwise healthy 6-year-old female patient with retinal venous thromboses. DESIGN: Observational case report. METHODS: A 6-year-old girl with poor vision in the left eye and preretinal hemorrhage underwent testing for infectious, autoimmune and embolic disease, diabetes, and hypertension. Testing for factor V Leiden and prothrombin G20210A mutations, homocysteine, anticardiolipin antibodies (ACAs), lupus anticoagulant, and functional assays for protein S, protein C, and antithrombin III were performed to detect a hypercoagulable state. No IRB approval was necessary. RESULTS: Only a positive lupus anticoagulant and moderately elevated ACA IgG were found. The ACA IgG was moderately elevated on repeat testing 18 months later. Laser to nonperfused retina caused some regression of retinal neovascularization. Aspirin was recommended to reduce the risk of future thromboses. CONCLUSIONS: Although uncommon, retinovascular thrombosis in children can occur in APA syndrome. Testing for ACA and lupus anticoagulant should be considered. PMID- 12654377 TI - Newly designed self-retaining contact lens for vitreous surgery. AB - PURPOSE: Development of a new self-retaining acrylic contact lens for vitreous surgery. DESIGN: A standard -60 diopters plano concave lens with the radius of curvature 6.7 mm (less than the cornea curvature) was modified with addition of four foot plates to facilitate stability and centration. METHODS: A drop of viscoelastic material placed between lens and cornea induces negative suction and helps retain the lens in position. CONCLUSIONS: This specially designed lens eliminates the need for suturing on the sclera or dependence on the assistant to provide placement. It provides a stable well-centered view of the fundus during vitreous surgery. PMID- 12654379 TI - Internal limiting membrane removal for macular detachment in highly myopic eyes. AB - PURPOSE: To present a treatment option for highly myopic eyes developing retinal detachment over a posterior staphyloma. DESIGN: Interventional case report. METHODS: Pars plana vitrectomy with peeling of the internal limiting membrane and gas tamponade; no laser retinopexy. RESULTS: Retinal reattachment with marked visual improvement. CONCLUSIONS: It appears that the rigid internal limiting membrane is responsible for the retina's inability to conform to the posterior staphyloma. Intraoperative findings suggest that this rigidity may also play a role in subsequent macular hole development. Internal limiting membrane removal should be considered in eyes with retinal detachment over a posterior staphyloma, whether or not a macular hole is present. PMID- 12654378 TI - Spontaneous resolution of a shallow detachment of the macula in a highly myopic eye. AB - PURPOSE: To report a case of pathologic myopia with shallow detachment of the macula and anatomic reattachment after spontaneous posterior vitreous separation. DESIGN: Observational case report. METHODS: A 66-year-old woman with pathologic myopia presented with decreased vision of 20/400 in her left eye. Biomicroscopy showed myopic chorioretinal changes, posterior staphyloma, and a shallow macular elevation. Optical coherence tomography was performed at the initial and follow up examinations. RESULTS: Initial optical coherence tomography revealed a retinal detachment, retinoschisis, and incomplete posterior vitreous detachment. One and one-half years after the initial visit the patient's best-corrected visual acuity improved to 20/200, and optical coherence tomography disclosed macular reattachment, release of vitreous adhesions, and restoration of normal retinal tomographic appearance. CONCLUSIONS: Spontaneous resolution of chronic shallow detachment of the macula in highly myopic eyes may occur and can be attributed to spontaneous posterior vitreous detachment. PMID- 12654380 TI - Neutrophil margination as a possible mechanism for verteporfin infusion associated pain. AB - PURPOSE: To investigate the changes in absolute neutrophil counts in patients with and without pain after verteporfin infusion. DESIGN: Experimental study. METHODS: The absolute neutrophil counts were obtained from the blood samples taken at the beginning of infusion and after the end of verteporfin infusion. RESULTS: Four patients in whom verteporfin infusion-related pain developed had a decrease in the absolute neutrophil counts from a preinfusion mean of 4,589/mm(3) to a postinfusion mean of 1,688/mm(3) while they had pain, as compared with preinfusion and postinfusion counts of 4,912/mm(3) and 4,111/mm(3), respectively, in five control patients having no pain, a difference in proportion that was significant (P =.027). CONCLUSIONS: Patients in this study with verteporfin infusion-associated pain developed neutropenia during infusion, most likely from drug-induced neutrophil margination. It is possible that medications preventing the transient neutropenia may help avert verteporfin infusion-associated pain. PMID- 12654382 TI - Frequency of epithelial debridement during diabetic vitrectomy. AB - PURPOSE: To survey active vitreoretinal surgeons to determine how often they intentionally debride the corneal epithelium during vitrectomy surgery for diabetic patients and to ascertain whether the selection of the surgical lens type influences the debridement rate. DESIGN: Observational retrospective case series. METHODS: Fifty-five vitreoretinal surgeons were asked to retrospectively report how many pars plana vitrectomies they performed in 1 year on diabetic eyes and in what percent of the cases was debridement of the cornea necessary. In a second query, they were asked to note which specific type of surgical lens system (hand-held irrigating, Landers sew-on, or Oculus BIOM noncontact) was used for their surgical intervention. RESULTS: The total number of diabetic vitrectomies performed in 1 year by the respondents was 8,002. The frequency of epithelial debridement was 17.4%, with a range of 0% to 90%. The use of irrigating contact lenses was associated with a significantly higher rate of debridement compared with the use of sew-on or BIOM noncontact lenses (23.5% vs 12.1%, P <.001). Regarding the 41 surgeons who indicated a specific type of lens used, we found that the debridement rate for infusion lenses was 23.8% compared with 13.0% for sew-on lenses and 15.6% for noncontact BIOM lenses. The difference between these groups was statistically significant (P <.025). CONCLUSIONS: Irrigating contact lenses appear to increase the need for epithelial debridement compared with other alternatives. Sew-on lenses with a viscoelastic cushion may provide the best corneal protection. PMID- 12654383 TI - Corneal perforation during laser in situ keratomileusis after hyperopic electrothermal keratoplasty. AB - PURPOSE: To report a corneal perforation during laser in situ keratomileusis (LASIK) after previous electrothermokeratoplasty. DESIGN: Interventional case report. METHODS: A 49-year-old man presented with primary hyperopia in the right eye and residual hyperopia after electrothermokeratoplasty in the left eye. His refraction was +4.00 in the right eye and +7.00 -3.00 x 135 degrees in the left eye, with a central pachymetry of 535 microm and 549 microm, respectively. Phacoemulsification with intraocular lens (IOL) insertion in the right eye and a two-step keratophacorefractive procedure with a piggyback IOL insertion and LASIK in the left eye were proposed. RESULTS: Postoperative refraction was -0.50 -0.50 x 150 degrees 20/20 in the right eye. Postphacoemulsification refraction was 4.75 -4.25 x 135 degrees in the left eye. Laser in situ keratomileusis was performed in the left eye, 4 months later, with uneventful astigmatic laser ablation. During the myopic ablation, a sudden outcome of aqueous humor in one of the temporal corneal scars was observed. CONCLUSIONS: Unpredictably thin areas after electrothermokeratoplasty may lead to unexpected corneal perforation during LASIK. The available pachymetry systems may be unreliable after electrothermal keratoplasty. PMID- 12654381 TI - Progressive outer retinal necrosis in immunocompetent patients treated initially for optic neuropathy with systemic corticosteroids. AB - PURPOSE: To report two cases of progressive outer retinal necrosis occurring in immunocompetent individuals after treatment with corticosteroids for presumed optic neuropathy. DESIGN: Observational case report. SETTING: University-based tertiary eye hospital. METHODS: Retrospective review of existing clinical records. RESULTS: Two patients were treated empirically with systemic corticosteroids for suspected inflammatory papillopathy. Subsequently, both were diagnosed with necrotizing herpetic retinitis with features of progressive outer retinal necrosis. Anterior chamber paracentesis confirmed varicella-zoster infection. Both patients were human immunodeficiency virus negative; one patient with rheumatoid arthritis was taking etanercept. Both became completely blind in one eye despite intensive treatment with antiviral medication intravenously and intravitreally. CONCLUSIONS: Progressive outer retinal necrosis is not confined to patients with underlying severe immunodeficiency, such as acquired immune deficiency syndrome. Initial treatment of acute, unexplained vision loss with systemic corticosteroids may lead to catastrophic visual loss in patients with evolving necrotizing herpetic retinopathy. PMID- 12654384 TI - Congenital adrenal hyperplasia due to 21-hydroxylase deficiency associated with bilateral keratoconus. AB - PURPOSE: To report a clinical association between congenital adrenal hyperplasia and keratoconus. DESIGN: Observational case report. METHODS: Two 25-year-old dizygotic female twins were retrospectively studied. Clinical history, hormonal serologic profile, complete ophthalmologic examination, and Holladay Diagnostic Summary variables were evaluated. In the course of a 15-month follow-up period, a progression of the corneal disease was observed. RESULTS: In both twins a diagnosis of nonclassical congenital adrenal hyperplasia, due to 21-hydroxylase deficiency, was established when they were 21 years old. In one subject, the computed corneal topographic analyses diagnosed an asymmetric keratoconus in both eyes. A progressive form of fruste central keratoconus was also documented in her sister. CONCLUSIONS: Congenital adrenal hyperplasia may be associated with keratoconus. An abnormal steroidogenic pathway, affecting the normal development of the cornea, could induce stromal abnormalities that lead to corneal ectasia. PMID- 12654385 TI - Sclerotic scattering illumination during phototherapeutic keratectomy for better visualization of corneal opacities. AB - PURPOSE: To report on a simple method to obtain a better view of corneal opacities during excimer laser phototherapeutic keratectomy (PTK). DESIGN: An interventional case report. METHODS: Phototherapeutic keratectomy was performed under sclerotic scattering illumination with a vitrectomy-endoilluminator at the limbus. RESULTS: During conventional PTK with diffuse illumination, the intraoperative observations of corneal opacities are not as good as that with a slit lamp. Conversely, subtle corneal opacities can be easily seen with sclerotic scattering illumination even under a surgical microscope. CONCLUSIONS: The use of sclerotic scattering illumination during PTK is valuable, because it provides a better view of opacified tissues and avoids excessive ablation. PMID- 12654386 TI - An unusual case of uveitis-glaucoma-hyphema syndrome. AB - PURPOSE: To report a case of uveitis-glaucoma-hyphema (UGH) syndrome in which anterior chamber paracentesis led to the diagnosis of sickle cell trait. DESIGN: Observational case report. METHODS: A 43-year-old Cuban pseudophakic male was seen multiple times over a 3-year period complaining of floaters and blurry vision in his left eye. He was noted to have an inferotemporally displaced posterior chamber intraocular lens and recurrent microhyphemas with elevated intraocular pressure (IOP) readings between 29 and 46 mm Hg with each episode. He was diagnosed with UGH syndrome. Posterior chamber intraocular lens explantation and anterior chamber washout was performed. The aqueous fluid was submitted for cytopathologic examination. RESULTS: Postoperatively, the patient's symptoms resolved and he had no further hemorrhages or elevated IOP readings. Cytopathology of the aspirate revealed sickled red blood cells. CONCLUSIONS: Microscopic examination of aqueous fluid can be a valuable tool in diagnosing ophthalmic manifestations of systemic disease. PMID- 12654387 TI - Three-dimensional eye movement analysis of superior oblique myokymia. AB - PURPOSE: To confirm the idea that sole contraction of the superior oblique muscle causes attacks of superior oblique myokymia (SOM). DESIGN: Observational case report. METHODS: A 43-year-old man presented with episodic monocular oscillatory eye movements. Three-dimensional eye movements were recorded using the magnetic search coil system and analyzed as rotation vector. The attacks of SOM consisted of intorsional, depressive, and abductive fast phases and their exponential decaying slow phases. RESULTS: Average direction of rotation axis for the fast phases while fixating straight ahead was 51.5 +/- 5.4 degrees from depression axis in vertical-torsional plane, 78.7 +/- 4.4 degrees from abduction axis in horizontal-torsional plane, and 76.3 +/- 4.3 degrees from abduction axis in horizontal-vertical plane (+/-SD, n = 50). Time constants of slow phases were less than 0.1 second. CONCLUSIONS: Quantitative three-dimensional analysis of SOM directly confirmed that the fast phases of SOM attacks were induced by the sole contraction of the superior oblique muscle. PMID- 12654388 TI - Abnormalities of scanning laser polarimetry associated with pituitary adenoma. AB - PURPOSE: To report abnormalities of retinal nerve fiber layer (RNFL) thickness using scanning laser polarimetry in patients with pituitary adenoma. DESIGN: Observational case reports. METHODS: Two patients with pituitary adenoma who had bitemporal visual field abnormalities were examined using scanning laser polarimetry. RESULTS: In Case 1, destruction of the normal two-peak pattern in the RNFL thickness was seen nasally by scanning laser polarimetry; this RNFL pattern corresponded well to visual field abnormalities. In Case 2, abnormalities in the RNFL pattern corresponded well to visual field defects not only in the superotemporal fields but also in the inferotemporal fields where the visual field defects were mild. CONCLUSIONS: Measurement of RNFL thickness using scanning laser polarimetry provides useful information in the diagnosis of chiasmal lesions such as pituitary adenomas. PMID- 12654389 TI - Unilateral Lisch nodules in the absence of other features of neurofibromatosis 1. AB - PURPOSE: To report a 14-year-old boy with unilateral Lisch nodules without any other diagnostic features of neurofibromatosis type 1. DESIGN: Observational case report. METHODS: A complete ophthalmologic examination and medical genetics examination was performed. RESULTS: By clinical examination, multiple Lisch nodules were identified in the left eye. CONCLUSIONS: Lisch nodules, in the absence of other diagnostic features of neurofibromatosis type 1, are an atypical finding. PMID- 12654390 TI - Ultrasound findings in orbital hydatid cysts. AB - PURPOSE: To describe the ultrasound features seen in orbital hydatid disease, which may confirm the diagnosis preoperatively. DESIGN: Interventional case series. METHODS: Nine patients of orbital hydatid cyst were evaluated preoperatively with ultrasound and computed tomography scans. The diagnostic features seen on ultrasound as well as the CT features are described. All patients underwent surgery to remove the cyst, and the diagnosis was confirmed surgically in all nine cases. RESULTS: We found that ultrasound was most helpful in diagnosing a hydatid cyst, even more so than a computed tomography scan, and we saw a diagnostic "double wall" sign in all cases, on ultrasound, a sign not previously described in orbital hydatids. CONCLUSIONS: It is difficult to diagnose an orbital hydatid cyst preoperatively. However, we found that the double wall sign could be elicited ultrasonically in all cases of hydatid cyst, if looked for carefully, and the diagnosis could be confirmed. PMID- 12654403 TI - The metaphor of patients as customers: implications for measuring satisfaction. AB - The use of satisfaction surveys in health care reflects the current tendency to think metaphorically of patients as "customers." This article reflects critically on the logic underlying this metaphor because metaphors are integral to the meaning of concepts. We argue that because the metaphor works differently when considering satisfaction with the process of care and satisfaction with treatment outcome, there are theoretical reasons for assessing these concepts from different perspectives. It seems reasonable to ask patients to rate their satisfaction with the processes of care or services received (e.g., hospital food, the physical environment) in much the same way they would rate services received at a repair shop or restaurant. When evaluating satisfaction with treatment outcome, however, the customer metaphor is problematic because the body is made an object when it is conceived of as the repairable possession of a customer. We conclude that measures of satisfaction with treatment outcome should be based on the assumption that rather than having bodies, people are embodied. Hence, the validity of satisfaction with treatment outcome would be enhanced by questions about psychologic, social, and experiental aspects of treatment outcome. PMID- 12654404 TI - Evaluating preference effects in partially unblinded, randomized clinical trials. AB - The ability of randomized, clinical trials (RCTs) to produce unbiased estimates of an intervention's specific (i.e., innate pharmacologic or physiologic) effect rests on at least two assumptions. First, other prognostic effects (such as disease severity or the psychologic effects of treatment) are assumed to be evenly distributed across treatment groups, and second, it is assumed that these other effects do not interact with the intervention's specific effect. This essay shows how the effect of patients' preferences for treatments may interact with an intervention's specific effect to bias the trial's results. Earlier mathematical descriptions of how preference effects may produce bias in nonblinded trials are extended to the case of (presumably) blinded RCTs. The extent to which preference effects may confer bias in partially unblinded placebo-controlled vs. active controlled drug trials are considered. Finally, methods for estimating preference effects in partially unblinded RCTs are provided. It is concluded that investigators could use these methods to quantify potential preference effects in partially unblinded RCTs, and thereby more accurately estimate the specific effects of treatments. PMID- 12654405 TI - Techniques for knowledge discovery in existing biomedical databases: estimation of individual aging effects in cognition in relation to dementia. AB - New interest is being expressed in the systematic application of modeling techniques to existing datasets. Under the rubric of Knowledge Discovery in Databases (KDD) large databases are being exploited for commercial and scientific purposes. This article reviews the development and applications of KDD techniques to dementia, using the longitudinal Canadian Study of Health and Aging dataset. KDD has demonstrated usefulness at the group level. For example, as in the course of functional impairment between Alzheimer's disease and no cognitive impairment suggest damage control-protection mechanisms for the former compared with noncompensated random accumulation of deficits for the latter. At the individual level, KDD suggests that more precise diagnosis seems possible as well as individual life expectancy prediction. Biomedical databases appear to hold the potential for novel insights when explored by systematic modeling. PMID- 12654406 TI - The validity of the diagnosis of acute myocardial infarction in routine statistics: a comparison of mortality and hospital discharge data with the Danish MONICA registry. AB - We analyzed the validity of the diagnosis of acute myocardial infarction (MI) in national registries by individual record linkage with the Danish MONICA registry. MI events were identified by record linkage between The National Hospital Registry and The National Death Registry. The sensitivity and specificity were estimated based on three different definitions of an MI event. For events with MI as the primary diagnosis or underlying cause of death the predictive value was 93.6% and sensitivity 77.6% when compared to DANMONICA definite or possible MI. Compared to DANMONICA, definite MI the predictive value was 75.0% and the sensitivity 92.8%. When secondary diagnosis and contributory cause of death were included, the sensitivity increased and the predictive value decreased only slightly. The combination of the national registries was found to be a valid and powerful tool for monitoring the population incidence of MI. PMID- 12654407 TI - The performance of instrumental activities of daily living scale in screening for cognitive impairment in elderly community residents. AB - A retrospective analysis of Short Orientation-Memory-Concentration (OMC) and Lawton's IADL data was performed to assess the association between instrumental activities of daily living (IADL) and a rating of cognitive impairment, and to test whether IADL measures can be used to screen for dementia. The study analyzed data from 1,095 elderly community residents who were regarded as potentially benefiting from care coordination. Three IADL items (telephone use, self medication, and handling finances) were statistically associated with cognitive impairment (OMC cutoff 10/11), independent of age and sex. An IADL indicator based on these items had only modest power in predicting cognitive impairment, its highest sensitivity being 0.71. Specificity was 0.75 at this point, but increased to 0.97 if higher indicator scores were used to define a positive result. In conclusion, the usefulness of an IADL indicator seems limited to ruling out further cognitive assessment rather than positively identify those with dementia. PMID- 12654408 TI - Quality of life assessment in the community-dwelling elderly: validation of the Assessment of Quality of Life (AQoL) Instrument and comparison with the SF-36. AB - Measurement of Health-Related Quality of Life (HRQoL) of the elderly requires instruments with demonstrated sensitivity, reliability, and validity, particularly with the increasing proportion of older people entering the health care system. This article reports the psychometric properties of the 12-item Assessment of Quality of Life (AQoL) instrument in chronically ill community dwelling elderly people with an 18-month follow-up. Comparator instruments included the SF-36 and the OARS. Construct validity of the AQoL was strong when examined via factor analysis and convergent and divergent validity against other scales. Receiver Operator Characteristic (ROC) curve analyses and relative efficiency estimates indicated the AQoL is sensitive, responsive, and had the strongest predicative validity for nursing home entry. It was also sensitive to economic prediction over the follow-up. Given these robust psychometric properties and the brevity of the scale, AQoL appears to be a suitable instrument for epidemiologic studies where HRQoL and utility data are required from elderly populations. PMID- 12654409 TI - Evaluating the quality of self-reports of hypertension and diabetes. AB - Increasingly, researchers and health specialists are obtaining information on chronic illnesses from self-reports. This study validates self-reports of two major health conditions, hypertension and diabetes, based on a recent survey in Taiwan (SEBAS 2000). These data, based on a large, nationally representative sample of respondents aged 54 and older, include both self-reported health information and a physical examination. Average blood pressure readings, laboratory measures of glycosylated hemoglobin, and information on whether the respondent was taking medication for hypertension or diabetes are used to validate respondents' reports of high blood pressure and diabetes. The resulting comparisons reveal that self-reports vastly underestimate the prevalence of hypertension, but yield a reasonably accurate estimate of the prevalence of diabetes. Significant correlates of the accuracy of the self-reports include age, education, time of the most recent health exam, and cognitive function. PMID- 12654410 TI - Systematic review of the epidemiologic and trial evidence of an association between antidepressant medication and breast cancer. AB - Biologic studies have suggested that antidepressant use may increase breast cancer risk. We conducted a systematic review of trials and controlled epidemiologic studies to assess this association. Pooled data from 31 primary efficacy drug company trials of fluoxetine suggested no increased risk but the short duration of these trials may have been insufficient to detect an association. In one prospective cohort study antidepressant use was associated with breast cancer, but this study was conducted among women attending for breast screening, and only limited data on antidepressant use were available. In a second large prospective drugs screening study no association was found between either amitriptyline or imipramine and breast cancer. In a large well-conducted retrospective cohort study there was no association between antidepressant use and breast cancer. A second retrospective cohort study was flawed, with exposure in those who developed breast cancer being measured over a shorter time period than in those who remained disease free. Two of four case-control studies found no association between antidepressant use and breast cancer after control for a number of potential confounding factors. We conclude that epidemiologic evidence does not support an association between antidepressant use and breast cancer. PMID- 12654411 TI - A retrospective cohort study of diabetes mellitus and antipsychotic treatment in the United States. AB - Treatment-emergent diabetes mellitus (DM) has been described for conventional and atypical antipsychotics. In our study, antipsychotic prescription claims from AdvancePCS's database were used to identify patients starting antipsychotic monotherapy. The relative risk of developing DM was determined using prescription claims for antidiabetic agents in the following cohorts: AdvancePCS general patient population, combined conventional antipsychotics, and combined atypical antipsychotics. Cox proportional hazards regression was used to adjust for differences in age, gender, and duration of antipsychotic exposure between cohorts in the estimation of risk of developing diabetes. Hazard ratios for developing DM in the combined conventional, combined atypical, and individual conventional and atypical antipsychotic treatment cohorts were greater than the AdvancePCS general patient population cohort. An increased risk of developing diabetes compared with the AdvancePCS general patient population was observed during treatment with conventional or atypical antipsychotics. PMID- 12654412 TI - Lifestyle factors and the risk of varicose veins: Edinburgh Vein Study. AB - The objective of this study was to determine the inter-relationships between a range of lifestyle factors and risk of varicose veins to identify which factors may be implicated in the etiology. An age-stratified random sample of 1566 subjects (699 men and 867 women) aged 18 to 64 years was selected from 12 general practices throughout Edinburgh. A detailed self-administered questionnaire was completed, and a comprehensive physical examination determined the presence and severity of varicose veins. The slightly higher age-adjusted prevalence of varicose veins in men than in women (39.7% versus 32.2%) was not explained by adjustment for an extensive range of lifestyle risk factors (male odds ratio [OR] 2.11, 95% confidence interval [CI] 1.51-2.96). In both sexes, increasing height showed a significant relationship with varicose veins (male OR 1.50, 95% CI 1.18 1.93 and female OR 1.26, 95% CI 1.01-1.58). Among women, body mass index was associated with an increased risk of varicose veins (OR 1.26, 95% CI 1.02-1.54). The current study casts doubt as to whether varicose veins occur predominantly in women. In addition, no consistent relationship with any lifestyle factor was shown. Self-reported evidence suggested a familial susceptibility, thereby warranting future genetic studies. PMID- 12654413 TI - Sex difference in hospitalization due to asthma in relation to age. AB - To describe the sex and age differences in asthma hospitalization among the Canadian population, we conducted an analysis based on a total of 9,486,173 hospital records in Canada for a 3-year period (1994/1995, 1995/1996, and 1996/1997), including 204,304 asthma patients and 288,977 asthma-related records. Asthma as one of the first five diagnoses, accounted for 3.0% of total hospitalizations, which was almost constant across the 3-year study period. The 3 year cumulative incidence of asthma hospitalization was substantially higher for young boys than girls, and it was reversed for adults. The incidence ratio for females vs. males for asthma hospitalization reached 2.8 for individuals 25 to 34 years of age, decreased gradually with increasing age, and then approached unity for those aged 80 years or more. The data suggest that sex is an important determinant for asthma, and the sex effect varies considerably over a life span. PMID- 12654414 TI - Education, poverty, and stroke incidence in whites and blacks: the NHANES I Epidemiologic Follow-up Study. AB - The purpose of this study was to test the hypothesis that educational attainment and poverty index are inversely associated with incidence of stroke in blacks and in whites. The NHANES I Epidemiologic Follow-up Study data were analyzed. We analyzed 2953 women and 2661 men with no history of stroke before baseline (1971 1975), using the incidence of stroke through 1992, years of education, and poverty index at baseline. In white men aged 45 to 74, Cox regression models showed an inverse age-adjusted association with education that did not attain statistical significance. In white women, those with 12 or more years of education had significantly lower age-adjusted risk of stroke compared with those with less than 8 years. A test for linear trend was significant when adjusting for age (P = 0.0005). In blacks, stroke risk was significantly lower in those with 8 or more years of education than in those with <8 years in adjusted models. In each group, Cox regression models showed an inverse, graded, age-adjusted association with poverty index that attained statistical significance. After controlling for multiple confounders and risk mediators, the association was diminished and nonsignificant. PMID- 12654415 TI - Insulin-mediated effects of alcohol intake on serum lipid levels in a general population: the Hisayama Study. AB - To determine whether the beneficial effects of alcohol on lipid concentrations are mediated by insulin levels, we performed a cross-sectional analysis in 2103 nondiabetic men and women aged 40 to 79 years from a general Japanese population in Hisayama. The multivariate-adjusted sum of fasting and 2-hour postloading insulin levels and the insulin resistance index significantly decreased with elevating alcohol intake levels in men (P < 0.01 for the trend) but not in women. No dose-response relations between alcohol intake and glucose levels were observed. In both sexes, high-density lipoprotein cholesterol (HDLC) significantly increased with elevated alcohol intake (P < 0.01), whereas total cholesterol and low-density lipoprotein cholesterol (LDLC) were inversely correlated with alcohol intake (P < 0.01). In contrast, triglycerides (TGs) levels in men showed a J-shaped relation to alcohol dose, with moderate drinkers (10-29 g/d) having the lowest levels. Estimates using regression models indicated that for men, 10% of the alcohol-induced increase in HDLC and 2% of the alcohol induced decrease in LDLC were insulin mediated. It was also estimated for male subjects that 36% of the reduction in TGs due to low to moderate alcohol intake was mediated by low levels of insulin and that this insulin-mediated pathway reduced the positive alcohol-TG relation by 13% in cases of moderate to heavy drinking. Our data suggest that regular alcohol consumption dose-dependently increased insulin sensitivity among male nondiabetics, but the insulin-mediated beneficial effects of alcohol on lipid concentrations were relatively small. PMID- 12654416 TI - Confidence intervals for adjusted NNEs: a simulation study. PMID- 12654417 TI - The ethics of sample size: the whole picture should be considered. PMID- 12654419 TI - High-dose-rate brachytherapy-. High-dose, high-tech, and high results. PMID- 12654420 TI - "Supplemental beam" and prostate brachytherapy: a simple answer to a complicated question? PMID- 12654421 TI - The challenge of predicting changes in pulmonary function tests after thoracic irradiation. PMID- 12654422 TI - Do "elderly fit" patients have less comorbidity? PMID- 12654423 TI - A comparison of surgical resection and stereotactic radiosurgery in the treatment of solitary brain metastases. AB - PURPOSE: To determine whether neurosurgery (NS) or stereotactic radiosurgery (RS) provided better local tumor control and enhanced patient survival. METHODS AND MATERIALS: Retrospective review of all solitary brain metastases (SBM) patients newly diagnosed at Mayo Clinic Rochester between 1991 and 1999. Eligible patients satisfied tumor size and SBM site criteria to qualify for both NS and RS. RESULTS: There were no significant differences between 74 NS and 23 RS patients in terms of baseline characteristics (age, gender, systemic disease type, systemic disease status, signs/symptoms at SBM presentation) or percent of patients who received whole brain radiotherapy. Median follow-up for alive patients was 20 months (range 0-106 months). There was no significant difference in patient survival (p = 0.15); the 1-year survival rate was 56% for the RS patients and 62% for the NS patients. Multivariate Cox regression analysis found that a significant prognostic factor for survival was a performance score of 0 or 1. There was a significant (p = 0.020) difference in local tumor control between NS and RS for solitary brain metastasis; none of the RS group had local recurrence compared to 19 (58%) of the NS group. CONCLUSION: The need for a Phase III study comparing these two techniques appears to be supported by the data from this study. PMID- 12654424 TI - A study on the radiation tolerance of the optic nerves and chiasm after stereotactic radiosurgery. AB - PURPOSE: To evaluate the risk of clinically significant radiation optic neuropathy (RON) for patients having stereotactic radiosurgery of benign tumors adjacent to the optic apparatus. METHODS AND MATERIALS: We reviewed the dose plans and clinical outcomes of 218 gamma knife procedures (215 patients) for tumors of the sellar and parasellar region (meningiomas, n = 122; pituitary adenomas, n = 89; craniopharyngiomas, n = 7 patients). Previous surgery or radiation therapy was performed in 156 (66%) and 24 (11%) patients, respectively. Median follow-up was 40 months (range 4-115). RESULTS: The median maximum radiation dose to the optic nerve was 10 Gy (range 0.4-16.0). Four patients (1.9%) developed RON at a median of 48 months after radiosurgery. All had prior surgery, and 3 of 4 had external beam radiotherapy (EBRT) in their management either before (n = 2) or adjuvantly (n = 1). The risk of developing a clinically significant RON was 1.1% for patients receiving 12 Gy or less. Patients receiving prior or concurrent EBRT had a greater risk of developing RON after radiosurgery (p = 0.004). CONCLUSION: RON occurred in less than 2% of our patients, despite the majority (73%) receiving more than 8 Gy to a short segment of the optic apparatus. Knowledge of the dose tolerance of these structures permits physicians to be more aggressive in treating patients with sellar or parasellar tumors, especially those with hormone-producing pituitary adenomas that appear to require higher doses to achieve biochemical remission. PMID- 12654425 TI - A phase I trial of etanidazole and hyperfractionated radiotherapy in children with diffuse brainstem glioma. AB - PURPOSE: To determine the toxicity and maximum tolerated dose of etanidazole administered concurrently with hyperfractionated radiation therapy (HRT) for children with brainstem glioma. METHODS AND MATERIALS: Eighteen patients with brainstem glioma were treated with etanidazole and HRT on a dose escalation protocol (Phase I trial) between 1990 and 1996. All patients had MRI confirmation of diffuse pontine glioma and signs/symptoms of cranial nerve deficit, ataxia, or long tract signs of <6 months' duration. Cervicomedullary tumors were excluded. Patients (median age: 8.5 years; 11 males, 7 females) received HRT to the tumor volume plus a 2-cm margin with parallel-opposed 6-15-MV photons. The total dose was 66 Gy in 44 fractions (1.5 Gy b.i.d., with at least 6 h between fractions) for the first 3 patients and 63 Gy in 42 fractions for the subsequent 15 patients. Etanidazole was administered as a rapid i.v. infusion 30 min before the morning fraction of HRT. Planned doses of etanidazole were 1.8 g/m(2) x 17 doses (30.6 g/m(2)) at Step 1 to a maximum of 2.4 g/m(2) x 21 doses (50.4 g/m(2)) at Step 8. Dose escalation was planned with 3 patients at each of the 8 levels. RESULTS: Three patients were treated at each dose level except Level 2, on which only 1 patient was treated. The highest dose level achieved was Level 7, which delivered a total etanidazole dose of 46.2 g/m(2). Two patients were treated at this level, and both patients experienced Grade 3 toxicity in the form of a diffuse cutaneous rash. Three patients received a lower dose of 42 g/m(2) (dose Level 6) without significant toxicity, and this represents the maximum tolerated dose (MTD). There were 23 cases of Grade 1 toxicity (10 vomiting, 5 peripheral neuropathy, 2 rash, 2 constipation, 1 weight loss, 3 others), 11 cases of Grade 2 toxicity (4 vomiting, 2 skin erythema, 2 constipation, 1 arthralgia, 1 urinary retention, 1 hematologic), and 4 Grade 3 toxicities (2 rash, 1 vomiting, 1 skin desquamation). Grade 2 or 3 peripheral neuropathy was not seen at any dose level. The median survival from the start of treatment was 8.5 months (range: 3-58 months). CONCLUSION: The MTD of etanidazole in children receiving HRT for brainstem glioma is 42 g/m(2), with cutaneous rash as the dose-limiting toxicity. This is in contrast to the adult experience, which demonstrates a 24% lower MTD of 34 g/m(2) limited by peripheral neuropathy. PMID- 12654426 TI - Determinants and prognoses of locoregional and distant progression in breast cancer. AB - PURPOSE: To describe locoregional and distant progression in a population-based breast cancer sample. METHODS AND MATERIALS: Between 1978 and 1998, the Munich Cancer Registry evaluated 14,429 patients. The mean follow-up of survivors was 8.3 years. Metastases (MET), local recurrence (LR), and lymph node recurrence (LNR) were considered as outcome measures. The prognostic factor for, and effects of, LR and MET were assessed multivariately by the Cox and dynamic Aalen models. RESULTS: The LR and MET rate increased with increasing tumor size, with the latter described by pT category. Distant MET occurred earlier than local progression. MET was recorded even earlier for MET alone. The mean time from diagnosis to MET for MET and LR was 54.9, 43.4, 29.4, and 24.7 months and for MET only was 36.5, 31.0, 22.6, and 12.9 months for pT1, pT2, pT3, and pT4, respectively. After MET, survival varied only slightly by pT stage; after LR, a more favorable prognosis, especially for pT1 and pT2, was evident. The prognosis after MET depended mainly on the MET location; 50% of patients with cerebral or nervous system MET survived <1 year and 50% of those with skeletal MET survived >2 years. In the Cox model, the relative risk of LR for MET was 3.0. In the Aalen model, after 30 months, when the hazard rates of MET began to decline, there was still an excess risk of MET after LR. CONCLUSION: This disease description highlights the importance of long-term observational studies. Empiric evidence that LR is both an indicator for, and in part a cause of, MET has been provided. In the future, the MET location should be reported. Variations in guidelines or health care systems that influence the time to MET and survival after MET through different diagnostic procedures should also be considered. PMID- 12654427 TI - Breast conservation rates-barriers between tertiary care and community practice. AB - PURPOSE: Low rates of breast conservation therapy (BCT) are reported in the southern United States. We evaluated the influence on BCT rates of opening a radiotherapy (RT) clinic at a community hospital in North Carolina. Before opening, RT was available 5 miles away at a tertiary care center. METHODS AND MATERIALS: A review of the pathology database of the community hospital identified patients who underwent definitive surgery for invasive breast malignancy or ductal carcinoma in situ between 1994 and 1995, and 1997 and 1998, before and after the opening of the RT clinic in 1996. From these data, the mode of therapy, mastectomy or BCT, was determined. The results were compared using logistic regression analysis. Surgical and RT physician staffing were unchanged throughout the study period. RESULTS: A total of 586 patients was evaluated. The BCT rate at the community hospital for 1994-1995 and 1997-1998 was 29% and 44%, respectively. On both univariate and multivariate logistic regression analysis, the era of treatment was statistically significant in its impact on the procedure performed (p <0.001). CONCLUSION: The use of BCT increased at a community hospital after the opening of an on-site RT facility, even though RT was available 5 miles away previously. PMID- 12654428 TI - Ten-year outcome after combined modality therapy for inflammatory breast cancer. AB - PURPOSE: To evaluate the long-term outcome of combined modality therapy for inflammatory breast cancer. METHODS AND MATERIALS: The data from 54 women treated between 1983 and 1996 for inflammatory breast cancer were analyzed. Patients with metastatic disease or disease progression on induction chemotherapy were excluded. Induction chemotherapy was given to 52 patients. Mastectomy was performed in 52 patients. Radiotherapy was delivered to the breast or chest wall and regional lymph nodes in all patients. The median follow-up for all patients was 5.1 years. RESULTS: The 5- and 10-year overall survival rate was 56% and 35%, respectively; the corresponding relapse-free survival rates were 49% and 34%. Patients with a pathologic complete response after chemotherapy with or without preoperative radiotherapy had better 5- and 10-year overall survival rates (65% and 46%, respectively) and 5- and 10-year relapse-free survival rates (59% and 50%, respectively) compared with patients without a pathologic complete response. Those patients had a 5- and 10-year relapse-free survival rate of 45% and 27%, respectively. Locoregional failure at 5 and 10 years was 8% and 19%, respectively. CONCLUSION: The outcomes for patients completing multimodality therapy compare favorably with published data; however, the exclusion of patients with progression during induction chemotherapy may account in part for these results. The pathologic complete response rate was found to be an important prognostic factor. Selected patients with inflammatory breast cancer have the potential for long-term survival. PMID- 12654429 TI - Risk of lymphedema after regional nodal irradiation with breast conservation therapy. AB - PURPOSE: To evaluate the risk factors for lymphedema in patients receiving breast conservation therapy for early-stage breast cancer. METHODS AND MATERIALS: Between 1982 and 1995, 727 Stage I-II breast cancer patients were treated with breast conservation therapy at Massachusetts General Hospital. A retrospective analysis of the development of persistent arm edema was performed. Lymphedema was defined as a >2-cm difference in forearm circumference compared with the untreated side. The median follow-up was 72 months. Breast and regional nodal irradiation (BRNI) was administered in 32% of the cases and breast irradiation alone in 68%. RESULTS: Persistent arm lymphedema was documented in 21 patients. The 10-year actuarial incidence was 4.1%. The median time to edema was 39 months. The only significant risk factor for lymphedema was BRNI. The 10-year risk was 1.8% for breast irradiation alone vs. 8.9% for BRNI (p = 0.001). The extent of axillary dissection did not predict for lymphedema even within the subgroups of patients defined by the extent of irradiation. Most patients underwent Level I or II dissection. In this subgroup, the lymphedema risk at 10 years was 10.7% for BRNI vs. 1.0% for breast irradiation alone (p = 0.0003). CONCLUSION: Nodal irradiation was the only significant risk factor for arm lymphedema in patients receiving breast conservation therapy for early-stage breast cancer. Our data suggest that this risk is low with Level I/II dissection and breast irradiation. However, even after the addition of radiotherapy to the axilla and supraclavicular fossa, the development of lymphedema was only 1 in 10, lower than generally recognized. PMID- 12654430 TI - Radiation-induced DNA damage and repair in lymphocytes from breast cancer patients and their correlation with acute skin reactions to radiotherapy. AB - PURPOSE: Repair of radiation-induced DNA damage plays a critical role for both the susceptibility of patients to side effects after radiotherapy and their subsequent cancer risk. The study objective was to evaluate whether DNA repair data determined in vitro are correlated with the occurrence of acute side effects during radiotherapy. METHODS AND MATERIALS: Breast cancer patients receiving radiation therapy after a breast-conserving surgery were recruited in a prospective epidemiologic study. As an indicator for clinical radiosensitivity, adverse reactions of the skin were recorded. Cryo-preserved lymphocytes from 113 study participants were gamma-irradiated with 5 Gy in vitro and analyzed using the alkaline comet assay. Reproducibility of the assay was determined by repeated analysis (n = 26) of cells from a healthy donor. A coefficient of variation of 0.3 was calculated. RESULTS: The various parameters determined to characterize the individual DNA repair capacity showed large differences between patients. Eleven patients were identified with considerably enhanced DNA damage induction, and 7 patients exhibited severely reduced DNA repair capacity after 15 and 30 min. Six patients were considered as clinically radiosensitive, indicated by moist desquamation of the skin after a total radiation dose of about 50 Gy. CONCLUSIONS: Using the alkaline comet assay as described here, breast cancer patients were identified showing abnormal cellular radiation effects, but this repair deficiency corresponded only at a very limited extent to the acute radiation sensitivity of the skin. Because impaired DNA repair could be involved in the development of late irradiation effects, individuals exhibiting severely reduced DNA repair capacity should be followed for the development of late clinical symptoms. PMID- 12654431 TI - Concurrent mitomycin C, 5-fluorouracil, and radiotherapy in the treatment of locally advanced carcinoma of the cervix: a randomized trial. AB - PURPOSE: This is a prospective, Phase III multicenter randomized trial to assess the effectiveness of concurrent intravenous mitomycin C, oral 5-fluorouracil (5 FU), and radiotherapy (RT) in locally advanced carcinoma of the cervix. METHODS AND MATERIALS: Between January 1988 and November 1994, 926 patients with locally advanced carcinoma of the cervix, FIGO Stage IIB-IVA, were entered into this study. The patients were randomized into four arms, as follows: Arm 1: conventional RT; Arm 2: conventional RT and adjuvant chemotherapy; Arm 3: conventional RT plus concurrent chemotherapy; Arm 4: conventional RT plus concurrent chemotherapy and adjuvant chemotherapy. Concurrent chemotherapy consisting of intravenous mitomycin C at 10 mg/m(2) was given on Days 1 and 29, and oral 5-FU at 300 mg/day was administered on Days 1-14 and 29-42 during RT. Adjuvant chemotherapy of 5-FU orally at 200 mg/day was given for three courses of 4 weeks, with a 2-week rest every 6 weeks. Six centers participated in the trial. RESULTS: The median follow-up time was 89 months. Acute side effects were generally higher in concurrent arms, but most of the patients tolerated the treatment well. Bone marrow toxicity was also higher in concurrent arms. The 5 year actuarial disease-free survival (DFS) was 48.2%, 54.1%, 64.5%, and 59.7% for arms 1, 2, 3, and 4, respectively. The pattern of failure revealed a significant increase in locoregional recurrence in the nonconcurrent chemoradiotherapy arm. The local recurrence was 25.5%, 20.6%, 14.3%, and 17.6% for arms 1, 2, 3, and 4, respectively. The metastatic rates were not significantly different in all four arms. At the time of analysis, there were no increases in late side effects, especially in gastrointestinal and genitourinary systems. CONCLUSIONS: Concurrent chemotherapy, mitomycin C, and 5-FU together with conventional RT showed an improved DFS rate when compared with conventional RT alone in patients with locally advanced carcinoma of the cervix. PMID- 12654432 TI - Assessing tumor hypoxia in cervical cancer by positron emission tomography with 60Cu-ATSM: relationship to therapeutic response-a preliminary report. AB - PURPOSE: Tumor hypoxia is associated with poor response to therapy. We have investigated whether pretreatment tumor hypoxia assessed by positron emission tomography (PET) with Cu-60 diacetyl-bis(N(4)-methylthiosemicarbazone) ((60)Cu ATSM) predicts responsiveness to subsequent therapy in cervical cancer. METHODS AND MATERIALS: Fourteen patients with biopsy-proved cervical cancer were studied by PET with (60)Cu-ATSM before initiation of radiotherapy and chemotherapy. (60)Cu-ATSM uptake was evaluated semiquantitatively by determining the tumor-to muscle activity ratio (T/M) and peak slope index of tumor tracer uptake. All patients also underwent clinical PET with F-18 fluorodeoxyglucose (FDG) before institution of therapy. The PET results were correlated with follow-up evaluation (14-24 months). RESULTS: Tumor uptake of (60)Cu-ATSM was inversely related to progression-free survival and overall survival (log-rank p = 0.0005 and p = 0.015, respectively). An arbitrarily selected T/M threshold of 3.5 discriminated those likely to develop recurrence; 6 of 9 patients with normoxic tumors (T/M < 3.5) are free of disease at last follow-up, whereas all of 5 patients with hypoxic tumors (T/M > 3.5) have already developed recurrence. Similar discrimination was achieved with the peak slope index. The frequency of locoregional nodal metastasis was greater in hypoxic tumors (p = 0.03). Tumor FDG uptake did not correlate with (60)Cu-ATSM uptake (r = 0.04; p = 0.80), and there was no significant difference in tumor FDG uptake between patients with hypoxic tumors and those with normoxic tumors. CONCLUSION: (60)Cu-ATSM-PET in patients with cervical cancer revealed clinically relevant information about tumor oxygenation that was predictive of tumor behavior and response to therapy in this small study. PMID- 12654433 TI - Estimating hypoxic status in human tumors: a simulation using Eppendorf oxygen probe data in cervical cancer patients. AB - PURPOSE: To define the minimal number of pO(2) measurements, with 90% sensitivity and 90% specificity, needed to categorize cervical tumors as either hypoxic or oxic. METHODS AND MATERIALS: Using Eppendorf oxygen probe data from our ongoing prospective trial, we simulated the measurement of tumor oxygenation with a smaller number of data points in 135 patients with cervical cancer. The hypoxic proportion, defined as the percentage of pO(2) values <5 mm Hg (HP5), was calculated for each tumor. Hypoxic tumors were defined as those with a median HP5 >50%, and tumors with normal oxygen levels as those with a median HP5 < or =50%. A small number of pO(2) measurements were randomly selected from the Eppendorf measurements in each tumor, or per Eppendorf track, and used to define the tumor as hypoxic or oxic. The sensitivity and specificity were calculated, considering the classification as given by the complete set of Eppendorf measurements as the reference standard. RESULTS: The probability of falsely classifying the tumor decreased as the selected number of pO(2) measurements per tumor increased, and at 16 measurements was approximately 10%. Adding additional measurements per tumor beyond 24 improved the ability to classify the tumor accurately only slightly. The probability of falsely classifying the tumor decreased as the pO(2) measurements per track increased. At five measurements per track, the probability of falsely classifying the tumor was approximately 9%. CONCLUSION: Approximately 20 measurements per tumor, or five measurements per track, using the Eppendorf pO(2) histograph, are sufficient to categorize cervical tumors as hypoxic or oxic. The results of this study will serve as a guide for research clinicians in the use of this and other systems in the assessment of tumor oxygenation in humans. PMID- 12654434 TI - Hyperfractionated radiotherapy with concurrent chemotherapy for para-aortic lymph node recurrence in carcinoma of the cervix. AB - PURPOSE: To evaluate efficacy, toxicity, and patterns of relapse in patients treated with hyperfractionated radiotherapy (HFRT) with concurrent chemotherapy for para-aortic lymph node (PALN) recurrence of cervical carcinoma. METHODS AND MATERIALS: Between September 1997 and October 2000, 12 cervical carcinoma patients with isolated PALN recurrence who had previously received radical or postoperative radiotherapy were treated with HFRT and concurrent chemotherapy. The initial FIGO stage was Stage IB in 4 (33%) patients, Stage IIA in 2 (17%), and Stage IIB in 6 (50%). The radiation field encompassed the gross recurrent PALN with the superior margin at the upper end of the T12 body and the inferior margin between L5 and S1. The fractionated dose was 1.2 Gy in 2 daily fractions, and the median total dose was 60 Gy. The weekly concurrent chemotherapy consisted of paclitaxel in 11 patients and cisplatin in 1. The median number of cycles of chemotherapy was 5. RESULTS: The latent period to PALN recurrence from the time of initial treatment for all patients ranged from 2 to 92 months (median: 12 months). One month after treatment, the clinical tumor response evaluated was complete in 33% (4/12) and partial in 67% (8/12). The 3-year overall survival rate and median survival were 19% and 21 months, respectively. The latent period to PALN recurrence was the only significant prognostic factor; the median survival of patients who relapsed in < or =24 months from the initial treatment of cervical carcinoma was 13 months vs. 45 months for those relapsed at >24 months (p = 0.026). Grade 3-4 hematologic toxicity developed in 2 patients. Six (50%) patients experienced Grade 2 nausea. There were no late gastrointestinal or neurologic complications during the follow-up period. Subsequent distant metastases after PALN treatment developed in 58% (7/12). CONCLUSION: HFRT of 60 Gy to PALN with concurrent chemotherapy could be regarded as an effective treatment modality without significant acute or late toxicity. Patients with a latent period >24 months until PALN recurrence had a more favorable survival rate than those with a latent period 2 mm). Postoperative radiotherapy was mandatory for surgery only patients with a positive CRM, but was not always administered and enabled us to compare local recurrence rates for patients with or without postoperative radiotherapy. Furthermore, the effect of preoperative radiotherapy was assessed in the different margin groups. RESULTS: Of 120 patients in the surgery-only group with a positive CRM, 47% received postoperative radiotherapy. There was no difference in the local recurrence rate between the irradiated and nonirradiated patients (17.3% vs. 15.7%, p = 0.98). Preoperative radiotherapy was effective in patients with a narrow CRM (0% vs. 14.9%, p = 0.02) or wide CRM (0.9 vs. 5.8%, p < 0.0001), but not in patients with positive margins (9.3% vs. 16.4%, p = 0.08). CONCLUSION: Preoperative hypofractionated radiotherapy has a beneficial effect in patients with wide or narrow resection margins, but cannot compensate for microscopically irradical resections resulting in positive margins. PMID- 12654444 TI - Do age and comorbidity impact treatment allocation and outcomes in limited stage small-cell lung cancer? a community-based population analysis. AB - PURPOSE: The effects of age and comorbidity on treatment and outcomes for patients with limited stage small-cell lung cancer (L-SCLC) are unclear. This study analyzes relapse and survival in a community-based population with L-SCLC according to age and comorbidity. METHODS: A retrospective review was performed on 174 patients with L-SCLC referred to the British Columbia Cancer Agency, Vancouver Island Centre, between January 1991 and December 1999. Patient and treatment characteristics, disease response, relapse, and survival were compared among three age cohorts: <65 years (n = 55, 32%), 65-74 years (n = 76, 44%), and > or =75 years (n = 43, 25%); and according to Charlson comorbidity scores 0, 1, and > or =2. Multivariate analysis was performed to identify independent prognostic factors associated with treatment response and survival. RESULTS: Patient factors that significantly differed with age were functional status classified by Eastern Cooperative Oncology Group performance status and number of comorbidities. Increasing age was significantly associated with fewer diagnostic scans. Combined modality chemoradiotherapy (CRT) was given in 86%, 66%, and 40% of patients ages <65, 65-74, and > or =75 years, respectively, (p <0.0001). Thoracic irradiation use was comparable among the age cohorts (p >0.05), but chemotherapy use varied significantly with less intensive regimens, fewer cycles, and lower total doses with advancing age (p <0.05). Prophylactic cranial irradiation (PCI) was used in 41 patients, only 3 of whom were age >70 years. Overall response rates to primary treatment significantly decreased with advancing age: 91%, 79%, and 74% in patients ages <65, 65-74, and > or =75 years, respectively (p = 0.014). Treatment toxicity and relapse patterns were similar across the age cohorts. Overall 2-year survival rates were significantly lower with advancing age: 37%, 22%, and 19% (p = 0.003), with corresponding median survivals of 17, 12, and 7 months among patients ages <65, 65-74, and > or =75 years, respectively. On multivariate analysis, age and Charlson comorbidity scores were not significantly associated with treatment response and survival. Independent prognostic factors favorably associated with survival were good performance status, normal lactate dehydrogenase, absence of pleural effusion, and > or =four cycles of chemotherapy. CONCLUSION: Increasing age was associated with decreased performance status and increased comorbidity. Older patients with L-SCLC were less likely to be treated with CRT, intensive chemotherapy, and PCI. Treatment response and survival rates were lower with advancing age, but this may be attributed to poor performance status and suboptimal treatment rather than age. PMID- 12654446 TI - Spontaneous and radiation-induced apoptosis in colorectal carcinoma cells with different intrinsic radiosensitivities: survivin as a radioresistance factor. AB - BACKGROUND: Spontaneous apoptosis has been shown to predict tumor response to radiochemotherapy in rectal cancer in vivo. It remains to be elucidated, however, which genetic profile determines whether a tumor is more or less prone to apoptosis. Recently, a novel member of the inhibitor of apoptosis protein family, designated survivin, was identified. We investigated the impact of survivin expression on tumor cell apoptosis in three colorectal cell lines of different intrinsic radiosensitivities. METHODS AND MATERIALS: Survivin protein expression was measured by Western blot analysis, and survivin mRNA expression by quantitative TaqMan reverse transcription polymerase chain reaction, both in untreated cell and after irradiation with 2 and 8 Gy. The expression profile was then correlated to spontaneous and radiation-induced apoptosis (Tunel-Assay, DAPI staining) in three colorectal cell lines of low (SW 480), intermediate (HCT-15), and high radiosensitivity (SW 48), as determined by the colony-forming assay. RESULTS: In vitro analysis revealed higher spontaneous and higher radiation induced apoptosis rates in the radiosensitive line (SW 48), as compared with the more resistant line (SW 480). In Western blot analysis and in TaqMan analysis, SW 480 was characterized by a higher spontaneous expression and a pronounced induction of survivin 48 h after irradiation, whereas survivin expression was low when untreated and not increased after irradiation in the most radiosensitive line SW 48. HCT-15 was intermediate, both with respect to the level of survivin mRNA and protein expression. CONCLUSION: The inverse correlation of survivin expression with spontaneous and radiation-induced apoptosis suggests that survivin is an important inhibitor of apoptosis in colorectal cancer cell lines. Analysis of survivin mRNA or protein expression may therefore provide predictive information on radio- and chemoresistance of individual colorectal tumors. PMID- 12654445 TI - Pulmonary function following high-dose radiotherapy of non-small-cell lung cancer. AB - PURPOSE: To study changes of pulmonary function tests (PFTs) after radiotherapy (RT) of non-small-cell lung cancer (NSCLC) in relation to radiation dose, tumor regression, and changes in lung perfusion. METHODS AND MATERIALS: Eighty-two patients with inoperable NSCLC were evaluated with PFTs (forced expiratory volume in 1 s [FEV(1)] and diffusion capacity [T(L,COc)]), a computed tomography (CT) scan of the chest, and a single photon emission CT (SPECT) lung perfusion scan, before and 3-4 months after RT. The reductions of PFTs and tumor volume were calculated. The lung perfusion was measured from pre- and post-RT SPECT scans, and the difference was defined as the measured perfusion reduction (MPR). In addition, the perfusion post-RT was estimated from the dose distribution using a dose-effect relation for regional lung perfusion, and compared with the pre-RT lung perfusion to obtain the predicted perfusion reduction (PPR). The difference between the actually measured and the PPR was defined as reperfusion. The mean lung dose (MLD) was computed and weighted with the pre-RT perfusion, resulting in the mean perfusion-weighted lung dose (MpLD). Changes of PFTs were evaluated in relation to tumor dose, MLD, MpLD, tumor regression, and parameters related to perfusion changes. RESULTS: In a multivariate analysis, the total tumor dose and MLD were not associated with reductions of PFTs. Tumor regression resulted in a significant improvement of FEV(1) (p = 0.02), but was associated with a reduction of T(L,COc) (p = 0.05). The MpLD and the PPR showed a significant (p = 0.01 to 0.04) but low correlation (r = 0.24 to 0.31) with the reduction of both PFTs. The other parameters for perfusion changes, the MPR and reperfusion were not correlated with changes in PFTs. CONCLUSION: The perfusion-related dose variables, the MpLD or the PPR, are the best parameters to estimate PFTs after RT. Tumor regression is associated with an improvement of FEV(1) and a decline of T(L,COc). Reperfusion was not associated with an improvement of global pulmonary function. PMID- 12654447 TI - Lack of interferon beta-induced radiosensitization in four out of five human glioblastoma cell lines. AB - PURPOSE: To investigate the potential of interferon beta to enhance the cytotoxic activity of ionizing irradiation against glioma cells, and to elucidate the possible mechanisms responsible for conflicting clinical results. METHODS AND MATERIALS: Five glioblastoma cell lines (U87MG, U118MG, U373MG, MO59K, MO59J) with different radiosensitivity and genetic background were used. Experiments were performed in exponentially growing cultures, and cell survival was measured by a colony-forming assay. Cells were incubated with natural interferon beta (n IFN-beta; 30-3000 IU/mL) for 24 h followed by single dose irradiation with 1 to 6 Gy of gamma-rays. RESULTS: Significant differences in n-IFN-beta sensitivity were found. The cell lines also differed in their radiation sensitivity, and there was no correlation between the n-IFN-beta and the radiation sensitivity. In three of five cell lines, the interaction of n-IFN-beta and irradiation was infra additive; in one cell line, it was additive. For MO59J cells only, which are NHEJ deficient, supra-additivity was observed. CONCLUSION: Our results confirm the remarkable heterogeneity that is characteristic of malignant glioma. The combined effect of n-IFN-beta and radiation was mostly infra-additive or additive; synergistic interaction might occur in tumor cells that already have acquired repair deficiencies because of their genetic instability, as shown for the MO59J cell line. PMID- 12654448 TI - Erythropoietin restores the anemia-induced reduction in radiosensitivity of experimental human tumors in nude mice. AB - PURPOSE: The effect of recombinant human erythropoietin (rhEPO) on the radiosensitivity of human tumor xenografts growing in anemic and nonanemic nude mice was studied. METHODS AND MATERIALS: Anemia was induced by total body irradiation ([TBI], 2 x 4 Gy) of mice before tumor implantation into the subcutis of the hind leg. The development of anemia was prevented by rhEPO (750 U/kg s.c.) given 3 times weekly starting 2 weeks before TBI. Fourteen days after fractionated TBI (tumor volume of approx. 40 mm(3)), single-dose irradiation of the tumor with varying doses was performed so that in full dose-response relationship for the probability of tumor cure was obtained. RESULTS: Radiation induced anemia (hemoglobin concentration [cHb] = 9.9 g/dl) led to a reduced radiosensitivity compared to controls [49.4 vs. 40.1 Gy radiation dose to control 50% of the tumors (TCD50)]. Upon rhEPO treatment for anemia prevention (cHb = 13.3 g/dl), the TCD50 was 39.8 Gy, illustrating restored radiosensitivity compared to anemic mice. CONCLUSION: These data provide further experimental evidence for restored radiosensitivity upon prevention of anemia with rhEPO. PMID- 12654450 TI - Experimental thermoradiotherapy in malignant hepatocellular carcinoma. AB - PURPOSE: The human liver is known to be a relatively radiosensitive organ that develops clinically relevant late radiation hepatitis subsequent to whole liver treatment with total doses above 30 Gy in conventional fractionation. Experimental data, as well as clinical series, have demonstrated that hyperthermia of solid tumors in addition to radiotherapy enhances tumor growth inhibition and tumor control probability. We therefore developed an experimental model for combined radiotherapy and hyperthermia of the liver in transplantable rat Morris hepatoma 3924A. METHODS AND MATERIALS: A cube of approximately 8 mm(3) was implanted subcapsularly into the middle liver lobe of 59 male syngenic ACI rats weighing approximately 180-200 g. On Day 16 after tumor implantation, irradiation of the tumor-bearing liver with either 0 Gy/25 Gy/35 Gy/45 Gy total dose in 10 fractions +/- hyperthermia (target temperature 40-42 degrees C) twice a week was initiated. Energy deposition was monitored by temperature probes in the liver and esophagus of the rats. Determination of tumor volume with magnetic resonance imaging was performed 2 to 5 weeks after the end of therapy. The tumor growth rates could be estimated for 44 rats. If the growth rate was positive (37 rats), the inverse of the growth rate was interpreted as the time to 10-fold tumor volume. Otherwise the maximum observation time was considered as a censored value in a parametric survival analysis. RESULTS: Intrahepatic temperature probes showed a temperature plateau of greater than 40 degrees C after 5 to 8 min subsequent to initiation of hyperthermia. The target temperatures could be maintained for at least 22 min > or =40 degrees C and 10 min > or =41 degrees C, respectively. Median plateau temperature in liver, esophagus, and epicutaneously was 41.2 degrees C (standard deviation [SD] 0.7 degrees C; range 38.2 to 43.3 degrees C), 40.4 degrees C (SD 1.08 degrees C; range 38.9 to 41.8 degrees C), and 40.8 degrees C (SD 0.8 degrees C; range 38.2 to 42.7 degrees C), respectively. Elevation of the temperature in the esophagus correlated with intrahepatic temperatures in the range of 39-42 degrees C, r = 0.957. The increase in time to 10-fold tumor volume for each step of irradiation dosage was by 34% (95% confidence interval [CI] 20% to 49%) without hyperthermia and by 60% (95% CI 47% to 80%) with hyperthermia (p < 0.0001). CONCLUSION: Treatment outcome after experimental percutaneous thermoradiotherapy in intrahepatically implanted Morris hepatoma 3924A was related to total dose of irradiation and concurrently administered regional hyperthermia. An increased radiosensitivity due to hyperthermia (<42 degrees C) has to be assumed. PMID- 12654449 TI - Enhancement of radiation therapy by tumor necrosis factor alpha in human colon cancer using a bispecific antibody. AB - PURPOSE: To overcome the systemic side effects of tumor necrosis factor alpha (TNFalpha) injected i.v., we used a bispecific antibody (BAb) directed against carcinoembryonic antigen (CEA) and TNFalpha to target this cytokine in human CEA expressing colorectal carcinoma treated simultaneously with radiation therapy (RT). METHODS, MATERIALS AND RESULTS: LS174T cell line was used to study the interaction of TNFalpha and radiation on clonogenic cytotoxicity. When TNFalpha (2500 U/mL) was added 12 h before RT, the surviving fraction at 2 Gy was 54% lower than that obtained with irradiation alone (0.23 vs. 0.42, respectively, p = 0.001). At 20%, 50%, or 70% survival, data points were within the envelope of additivity. Concerning in vivo experiments, RT as a single agent slowed tumor progression as compared with the control group (p = 0.027), whereas TNFalpha, BAb, or BAb + TNFalpha had no effect. BAb + TNFalpha + RT combination enhanced the delay for the tumor to reach 2000 mm(3) as compared with RT alone (p = 0.033, for BAb + TNFalpha + RT group vs. RT group). CONCLUSION: These results suggest that TNFalpha in combination with BAb and RT may be beneficial for the treatment of locally advanced colorectal cancer. PMID- 12654451 TI - Can PET provide the 3D extent of tumor motion for individualized internal target volumes? A phantom study of the limitations of CT and the promise of PET. AB - PURPOSE: To characterize the limitations of fast, spiral computed tomography (CT) when imaging a moving object and to investigate whether positron emission tomography (PET) can predict the internal target volume (ITV) and ultimately improve the planning target volume (PTV) for moving tumors. METHODS AND MATERIALS: To mimic tumors, three fillable spheres were imaged while both stationary and during periodic motion using spiral CT and PET. CT- and PET-imaged volumes were defined quantitatively using voxel values. Ideal PTVs for each scenario were calculated. CT-based PTVs were generated using margins of 7.5, 10, and 15 mm to account for both organ motion and setup uncertainties. PET-based PTVs were derived with the assumption that motion was captured in the PET images and only a margin (7.5 mm) for setup errors was necessary. Comparisons between CT based and PET-based PTVs with ideal PTVs were performed. RESULTS: CT imaging of moving spheres resulted in significant distortions in the three-dimensional (3D) image-based representations, and did not, in general, result in images well representative of either moving or stationary spheres. PET images were similar to the ideal capsular shape encompassing the sphere and its motion. In all cases, CT imaged volumes were larger than that for the stationary sphere (range of excess volume from 0.4 to 29 cm(3) for stationary volumes of 2.14 to 172 cm(3)), but smaller than that for the true motion volume. PET-imaged volumes were larger than the true motion volume (difference from ideal ranged from 3 to 94 cm(3) for motion volumes of 1.2 to 243 cm(3)) and much larger than the stationary volume. Using CT data, geographic miss of some part of the ideal PTV occurred for 0 of 24 cases, 11 of 24 cases, and 18 of 24 cases using a 15-mm, 10-mm, and 7.5-mm margin, respectively. Geographic miss did not occur in any case for the PET-based PTV. The amount of "normal tissue" included in CT-based PTVs was dramatically greater than that included in PET-based PTVs. CONCLUSION: Fast CT imaging of a moving tumor can result in poor representation of the time-averaged position and shape of the tumor. PET imaging can provide a more accurate representation of the 3D volume encompassing motion of model tumors and has potential to provide patient-specific motion volumes for an individualized ITV. PMID- 12654452 TI - Are multiple CT scans required for planning curative radiotherapy in lung tumors of the lower lobe? AB - PURPOSE: Lung tumors located in the lower lobe are the most mobile. Multiple computed tomographic (CT) scans, which had been performed for radiotherapy planning, were analyzed to determine the minimal number of required scans. METHODS AND MATERIALS: Six spiral CT scans (3 rapid and 3 slow) from 7 such patients were coregistered. Reproducibility of target volumes was defined as the ratio between the overlapping and encompassing volume (COM/SUM) from scans derived using one technique. Volumetric and dosimetric analyses were performed. RESULTS: Slow CT scans generated larger and more reproducible target volumes than rapid planning scans, with a mean COM/SUM ratio of 71.9 +/- 8.7% and 58.0 +/- 12.7%, respectively. When only a single slow CT scan was used for planning, the addition of a symmetrical 3D margin of 5 mm ensured 99% coverage of the "optimal" target volume, which was derived from summation of target volumes from all six scans. CONCLUSION: Planning target volumes (PTVs) derived from a single slow CT scan plus a 5-mm margin covered the "optimal" PTVs generated from six scans. Although these "slow PTVs" were larger, the increase in V(20) (the volume of lung tissue receiving a dose > or = 20 Gy) was limited. This indicates that only two CT scans, i.e., a full rapid scan of the entire thorax and a limited slow scan, are necessary for treatment planning in peripheral lung cancers. PMID- 12654453 TI - Patterns of patient movement during frameless image-guided radiosurgery. AB - PURPOSE: Image-guided radiosurgery aligns the treatment beam to the target site by using a radiographic imaging system to locate anatomic landmarks associated with the treatment target. Because the procedure is performed without a rigid frame, the precision of dose alignment can be affected by patient movement. Movement is limited by noninvasive restraints and compensated by remeasuring the target position at short intervals throughout treatment and then realigning the beam. Frameless image-guided radiosurgery has been used at our institution to treat 250 cranial, 23 spinal, 9 lung, and 3 pancreas cases involving malignant and benign tumors as well as vascular malformations. We have analyzed the target position records for all of these cases to assess the frequency, magnitude, and case-by-case patterns of patient movement. METHODS AND MATERIALS: The position of the treatment site during image-guided radiosurgery was measured at approximately 1-2-min intervals, on average, using orthogonal amorphous silicon X-ray cameras and an image registration process that determined all six degrees of freedom in the target's position. The change in position from one measurement to the next was indicative of patient movement. RESULTS: The treatment site position along each axis of translation was observed to vary by an average of 0.45 mm for the cranium, 0.53 mm for the cervical spine, 0.53 mm for the lumbar and thoracic spine, 1.06 mm for the lung, and 1.50 mm for the pancreas. Half of all cranial cases showed systematic drifting of the target away from the initial setup position. CONCLUSION: Using noninvasive restraints and supports, short-term movement of the head and spine during image-guided radiosurgery was limited to a radius of 0.8 mm, which satisfies the prevailing standard for radiosurgical dose alignment precision, but maintaining this margin of error throughout a treatment fraction requires regular monitoring of the target site's position. PMID- 12654454 TI - Quantifying the degree of conformity in radiosurgery treatment planning. AB - PURPOSE: To compare different parameters used to quantify the quality of a treatment plan and to evaluate the dose conformity and coverage clinically achieved using gamma knife radiosurgery. METHODS AND MATERIALS: Various existing parameters for coverage and conformity are reviewed. Additionally, a modified conformity index (CI) has been defined as the ratio of the volume within the target irradiated to at least the prescription isodose over the total volume enclosed by the prescription isodose. These parameters are calculated for all the 551 evaluable patient treatment plans. RESULTS: The median CI for all targets is 0.75, with a median target coverage of 94.6%. Regardless of the conformity parameter chosen, the conformity is seen to vary depending on the type of tumor and its location, reflecting the treatment planning philosophy. For tumors with volumes smaller than about 1 cm(3), the conformity parameter is also seen to be dependent on the target volume. CONCLUSION: With gamma knife radiosurgery, it is possible to achieve highly conformal dose distributions. A single parameter for the quantification of a plan, though desirable, is not realistic, because of the competing components of high dose to the target and low dose to normal tissue. Thus, we propose the use of the CI, together with the target volume coverage. PMID- 12654455 TI - A three-field breast treatment technique with precise geometric matching using multileaf collimator-equipped linear accelerators. AB - PURPOSE: Many authors have studied the problems associated with the three-field breast treatment, yet the proposed solutions present their own difficulties. This study presents a technique that overcomes these difficulties, reduces scatter to the contralateral breast, and improves setup reproducibility. METHODS AND MATERIALS: Patients are set up with both arms raised superiorly on a breast board. A precise field-match is achieved by rotating the couch and collimator of the tangents, while the supraclavicular field is half-beam blocked using an independent jaw. The posterior borders of the tangents are conformally defined by multileaf collimation. Measurements were performed to verify the field matching and evaluate scatter doses. RESULT: A smooth dose transition was found at the match line at all depths. Corner blocks and lower wedges were not used, which reduced the scatter to the contralateral breast compared with our prior technique. CONCLUSION: The technique achieves a precise match while removing constraints on the tangents' length and decreasing scatter dose. Procedures for simulation, planning, and treatment have been devised, along with a new patient setup routine incorporating orthogonal setup films and tattoos. This technique has been successfully implemented in routine treatment since September 2001. A program calculating the setup parameters is available at our website. PMID- 12654456 TI - Evaluation of rapid dose map acquisition of a scanning liquid-filled ionization chamber electronic portal imaging device. AB - PURPOSE: We evaluated the performance of a new dosimetry module in the LC250 scanning liquid-filled ionization chamber (SLIC) electronic portal imaging device (EPID) for intensity-modulated radiotherapy (IMRT) verification. This module permits one to convert EPID readings to two-dimensional (2D) maps of IMRT dose rate in real time, and to integrate them over time to produce a profile of accumulated dose for treatment verification. METHODS AND MATERIALS: The EPID was calibrated using an iterative procedure, from which a lookup table for dose integration was generated and transferred to the image-acquisition hardware. To evaluate the EPID's integration capability, we investigated the linearity of imaging time (vs. monitor unit [MU]) and integrated dose (vs. planned dose) for static and IMRT fields, in both standard ( approximately 2.7 s/image) and fast ( approximately 1 s/image) synchronous acquisition modes (S- and F-modes). We also compared the EPID-measured profiles with that measured using film and ionization chamber, or calculated from the treatment planning system. For the EPID's patient dose verification capability, we compared the integrated central-axis (CAX) dose with the planned dose for 25 prostate IMRT fields. We also compared the measured relative profiles with the planned ones using a linear regression model, which returns an index sigma (root mean squared error) for the goodness of fit. We identified errors that are either associated with the timing of the EPID-start delay and end truncation, or with the integration process-detector memory effects (decrease in detector's sensitivity with time during the fast continuous acquisition) and beam hold-off effects (the withholding of linac beam pulses when multileaf collimator leaves are not in the correct positions). The CAX doses of static fields were corrected using the ratio of the irradiation time to the imaging time. A linear decay model was proposed to correct the detector memory effect. To investigate the beam hold-off effect, we verified the relative profiles of a five-field prostate IMRT plan for five different MU settings, and correlated the goodness of fit with the percent of beam hold-off. RESULTS: The imaging time is linearly proportional to the given MU with a slope of 0.250 MU/s (ideal slope is 0.250 MU/s) and a R(2) = 1.0. Although the R(2) of the linearity for the measured vs. planned dose is 1.0 for both modes, only the slopes for the S-mode are within 3% of unity. The slopes for the F-mode deviate from unity due to detector memory effects, and are accurately corrected using the linear decay model. The EPID measured profiles agree well (within 2.0%) with the planned dose and profiles for both modes. For the CAX dose of the 25 IMRT fields, the S-mode is within 2% of the planned dose, whereas the F-mode is off significantly (>3%) if not corrected for detector memory effects. For the relative profile verification, lower MU always produces higher sigma for the same mode. The F-mode is more accurate than the S-mode for the same MU; however, the improvement is not proportional to the difference in imaging speed. Analysis of the correlation of the goodness of fit with the percent of beam hold-off indicates that the accuracy of profile verification for the F-mode is predominantly determined by the beam hold-off effect for lower MU. CONCLUSION: The S-mode of LC250 combined with a large MU can be used for the pretreatment verification of IMRT beam delivery with a significant reduction of processing time and computer resources in comparison to off-line processing. Real-time verification during treatment requires the F mode. Although the detector memory effects encountered in the F-mode can be compensated using the proposed linear decay model, sufficient accuracy for real time verification requires a resolution of the beam hold-off problem. PMID- 12654457 TI - Design optimization of intraoperative radiotherapy cones. AB - PURPOSE: Electron intraoperative cones (EIORCs) commonly used for intraoperative radiation therapy (IORT) often generate high-dose regions at superficial depths. This study was performed to optimize the use of rings in the EIORC design that reduces the high-dose region while minimizing the loss of the treatment volume at the prescribed depth. METHODS AND MATERIALS: Monte Carlo simulations were performed to study the dosimetry properties of various EIORC designs. Simulations were conducted with EIORCs of various internal radii, lengths, and material compositions irradiated by available electron beam energies. The data were analyzed in terms of volume receiving > 105% and < 90% of the prescription dose, respectively. RESULTS: The high-dose volume increases with the EIORC size and the electron beam energy. The use of a ring inside the EIORC reduces the 105% dose volume but also increases the sub-90% volume. The degree of change of these volumes depends on the ring thickness and position. CONCLUSION: The optimal ring position is about 10 cm from the bottom of the EIORC, regardless of the EIORC material, geometry, or electron energy. The optimal thickness of the ring is dependent on its material composition, the beam energy, and the preferred compromise between a uniform dose profile and a loss of treatment volume. PMID- 12654458 TI - In regard to Dobelbower RR, Simon Kramer 1919-2002: physician, teacher, pioneer, scientist, leader. IJROBP 2000;54:633-634. PMID- 12654459 TI - In regard to Zlotecki et al., the results of radiation therapy provide data to limit indications for surgery in aggressive fibromatosis in adults. IJROBP 2002;54:177-181. PMID- 12654460 TI - In regard to Rosenman et al., high-dose conformal radiotherapy for treatment of stage III A/B non-small-cell lung cancer: technical issues and results of a phase I/II trial. IJROBP 2002;54:348-356. PMID- 12654462 TI - In regard to hyperfractionation for non-small-cell lung cancer: fire, ready, aim! PMID- 12654464 TI - In regard to Pollack et al., prostate cancer radiation dose response: results of the M. D. Anderson phase III randomized trial. IJROBP 2002;53:1097-1105. PMID- 12654466 TI - Dietary flavonoids fail to suppress F2-isoprostane formation in vivo. AB - Dietary antioxidants, including alpha-tocopherol (alpha-TOH) and polyphenolic flavonoid compounds, have been the subject of much research interest, but few studies have investigated interactions between these two antioxidants in vivo. We have conducted a feeding study to determine if supplementation with dietary flavonoids or polyphenol-containing compounds will provide antioxidant protection in tocopherol-deficient animals or exceed the antioxidant protection provided by alpha-TOH alone, using the sensitive and specific measure of lipid peroxidation, F2-isoprostanes. Seventy-two male Sprague Dawley rats were divided into 12 treatment groups to receive either alpha-TOH-sufficient or -deficient AIN93-G diet supplemented with one of five compounds: 0.5% quercetin, catechin, or epicatechin; or 1% cocoa powder or lignin. The fat source was polyunsaturated oil, increased from 7 to 11.05% (w/w with diet) to maximize lipid peroxidation while staying within a physiological range. After 7 weeks of treatment, animals were sacrificed with plasma and hearts analyzed to determine differences in F2 isoprostane levels. None of the treatment compounds significantly decreased plasma or heart F2-isoprostanes compared to the control beyond the significant protection displayed by alpha-tocopherol. We conclude that under these experimental conditions, quercetin, catechin, and epicatechin do not suppress lipid peroxidation in vivo. PMID- 12654467 TI - Exercise, antioxidants, and HSP72: protection against myocardial ischemia/reperfusion. AB - Endurance exercise is associated with protection against myocardial ischemia/reperfusion (I/R) injury and has been shown to increase heat shock protein 72 (HSP72). Dietary antioxidants have also been reported to decrease I/R induced injury. Because exercise and antioxidants may provide cardioprotection via different mechanisms, combining these countermeasures could provide additive protection. Alternatively, because exercise-induced oxidant production may promote expression of HSP72, antioxidants could attenuate exercise-induced HSP72 expression and decrease exercise-related cardioprotection. These experiments examined the individual and combined effects of exercise and antioxidants on myocardial I/R injury (in vivo). Rats receiving a mixed antioxidant diet or control diet were assigned to exercise or sedentary groups and randomized to receive: (i) short I/R (myocardial stunning), (ii) long I/R (myocardial infarction), or (iii) sham surgery. Antioxidants significantly increased total antioxidant capacity and attenuated exercise-related HSP72 accumulation. Nonetheless, during short I/R, exercise-trained animals demonstrated improved left ventricular developed pressure (LVDP), independent of diet. Further, antioxidants alone resulted in improved LVDP. Finally, compared to control diet/sedentary animals, both exercise groups (control and antioxidant diets) and the antioxidant diet/sedentary group sustained smaller infarctions. We conclude that exercise and antioxidants can independently provide protection against myocardial contractile dysfunction and infarction, and the combination of these two strategies does not enhance or inhibit the protection observed with each individual countermeasure. PMID- 12654468 TI - Resveratrol, a red wine constituent polyphenol, prevents superoxide-dependent inflammatory responses induced by ischemia/reperfusion, platelet-activating factor, or oxidants. AB - Moderate consumption of red wine has been shown to exert cardioprotection against ischemia/reperfusion. Because oxidant-dependent leukocyte infiltration plays a critical role in ischemia/reperfusion-induced tissue injury, we hypothesized that resveratrol, a red wine constituent polyphenol would attenuate postischemic leukocyte recruitment and subsequent endothelial dysfunction. Intravital microscopic approaches were used to quantify leukocyte/endothelial cell interactions and venular protein leakage in rat mesenteries exposed to either 20 min ischemia and 60 min reperfusion (I/R), oxidants generated by the reaction of hypoxanthine and xanthine oxidase (HX/XO), platelet-activating factor (PAF), or leukotriene B4 (LTB4). I/R or HX/HX produced marked increases in the number of adherent (LA) and emigrated (LE) leukocytes, which were associated with significant increases in venular albumin leakage (VAL). Intravenous administration of resveratrol or superoxide dismutase (SOD) attenuated these increases in LA, LE, and VAL. Superfusion of the mesentery with PAF or LTB4 also markedly increased LA, LE, and VAL. While resveratrol attenuated the proinflammatory effects of PAF, LTB4-induced changes were not affected by resveratrol. Resveratrol prevents leukocyte recruitment and endothelial barrier disruption induced by a number of superoxide-dependent proinflammatory stimuli, including I/R, HX/XO, or PAF. These salutary effects appear to be related to the antioxidant properties of resveratrol and contribute to the cardioprotective actions associated with consumption of red wine. PMID- 12654469 TI - Oxidants and antioxidants affect the expression of glycodelin. AB - Glycodelin is a glycoprotein that has immunosuppressive activity. We have shown that K562 cells, hematopoitic progenitor cells, are capable of synthesizing glycodelin peptide (Gp) and, perhaps, contribute to Gp in tissues. In addition, several reproductive and nonreproductive tissues themselves are capable of synthesis of glycodelin. In this study, we report that lipid peroxides induce the synthesis of Gp. Antioxidants vitamin E and pyrrolidine dithiocarbamate (PDTC) and antioxidizing enzymes catalase and superoxide dismutase (SOD) effectively blocked phorbol myristate acetate- (PMA-) and lyso phosphatidic acid- (LPA-) induced synthesis of Gp. Dioctanoin (a mimic of diacylglycerol) activated Gp synthesis, and an inhibitor of protein kinase C (PKC) downregulated the response. Based on these observations, we postulate that oxidants by way of PKC might potentiate the angiogenic process. PMID- 12654470 TI - Expression of major HDL-associated antioxidant PON-1 is gender dependent and regulated during inflammation. AB - Paraoxonase 1, an HDL-associated enzyme that confers antioxidant activity on HDL, and its activity in serum have been correlated with protection against atherosclerosis, an oxidative disease. However, serum PON-1 activity is highly variable and its regulation is complex, involving both genetic and environmental factors. It is influenced by gender and inflammation, two important factors in atherosclerosis. Serum PON-1 activity has been shown to be lower in male mice and is decreased in male Syrian hamster during inflammation. Here we show that male mice had lower hepatic PON-1 mRNA that increased by 170% after castration. Our data also suggested that this effect was testes but not plasma testosterone dependent. Ovariectomy had no effect on PON-1 mRNA in female mice. LPS caused hepatic PON-1 mRNA to decrease further in male mice, and to increase moderately in female mice. Anti-inflammatory dexamethasone enhanced PON-1 mRNA level by 2 fold in male and female LPS-treated mice, and increased PON-1 expression by 8 fold in Hepa cell, a mouse hepatoma cell line. Therefore, antioxidant PON-1 is regulated at the mRNA level in a gender-specific manner by proinflammatory LPS and anti-inflammatory dexamethasone. PMID- 12654471 TI - Immunochemical detection of hemoglobin-derived radicals formed by reaction with hydrogen peroxide: involvement of a protein-tyrosyl radical. AB - To investigate the involvement of a hemoglobin radical in the human oxyhemoglobin (oxyHb) or metHb/H2O2 system, we have used a new approach called "immuno-spin trapping," which combines the specificity and sensitivity of both spin trapping and antigen:antibody interactions. Previously, a novel rabbit polyclonal anti DMPO nitrone adduct antiserum, which specifically recognizes protein radical derived nitrone adducts, was developed and validated in our laboratory. In the present study, the formation of nitrone adducts on hemoglobin was shown to depend on the oxidation state of the iron heme, the concentrations of H2O2 and DMPO, and time as determined by enzyme-linked immunosorbent assay (ELISA) and by Western blotting. The presence of reduced glutathione or L-ascorbate significantly decreased the level of nitrone adducts on metHb in a dose-dependent manner. To confirm the ELISA results, Western blotting analysis showed that only the complete system (oxy- or metHb/DMPO/H2O2) generates epitopes recognized by the antiserum. The specific modification of tyrosine residues on metHb by iodination nearly abolished antibody binding, while the thiylation of cysteine residues caused a small but reproducible decrease in the amount of nitrone adducts. These findings strongly suggest that tyrosine residues are the site of formation of the immunochemically detectable hemoglobin radical-derived nitrone adducts. In addition, we were able to demonstrate the presence of hemoglobin radical-derived nitrone adducts inside red blood cells exposed to H2O2 and DMPO. In conclusion, our new approach showed several advantages over EPR spin trapping with the anti DMPO nitrone adduct antiserum by demonstrating the formation of tyrosyl radical derived nitrone adduct(s) in human oxyHb/metHb at much lower concentrations than was possible with EPR and detecting radicals inside RBC exposed to H2O2. PMID- 12654472 TI - Structures of (-)-epicatechin glucuronide identified from plasma and urine after oral ingestion of (-)-epicatechin: differences between human and rat. AB - (-)-epicatechin is one of the most potent antioxidants present in the human diet. Particularly high levels are found in black tea, apples, and chocolate. High intake of catechins has been associated with reduced risk of cardiovascular diseases. There have been several reports concerning the bioavailability of catechins, however, the chemical structure of (-)-epicatechin metabolites in blood, tissues, and urine remains unclear. In the present study, we purified and elucidated the chemical structure of (-)-epicatechin metabolites in human and rat urine after oral administration. Three metabolites were purified from human urine including (-)-epicatechin-3'-O-glucuronide, 4'-O-methyl-(-)-epicatechin-3'-O glucuronide, and 4'-O-methyl-(-)-epicatechin-5 or 7-O-glucuronide, according to 1H- and 13C-NMR, HMBC, and LC-MS analyses. The metabolites purified from rat urine were 3'-O-methyl-(-)-epicatechin, (-)-epicatechin-7-O-glucuronide, and 3'-O methyl-(-)-epicatechin-7-O-glucuronide. These compounds were also detected in the blood of humans and rats by LC-MS. The presence of these metabolites in blood and urine suggests that catechins are metabolized and circulated in the body after administration of catechin-containing foods. PMID- 12654473 TI - Effect of catechin O-methylated metabolites and analogues on human LDL oxidation. AB - The effects of catechin metabolites and methylated analogues on LDL oxidation were studied in vitro using either a water-soluble initiator or copper ions to induce lipid peroxidation. Direct addition of catechin O-methylated analogues to the oxidation mixture led to a clear protective effect during lag phase and for the metabolites during both lag and propagation phases. The structure-activity relationships obtained with these selectively O-methylated compounds allowed determination of catechin active moietie: the catechol B-ring. Based on physical chemical studies, these results suggest that the mechanism implied in the scavenging properties of flavan-3-ols is not only hydrogen transfer, as generally described, but mainly an electronic transfer from the phenolate, and that 3'- and 4'-O-methylcatechin seem, moreover, to act as amphiphilic chain-breaking antioxidants. However, the plasma concentration of flavan-3-ols necessary to protect LDL is far greater than those usually found in human plasma. Therefore, the data do not support a direct physiological relevance of flavan-3-ols as antioxidants in lipid processes. Future research should focus on other effects besides simple antioxidant ones. PMID- 12654474 TI - Detection of endothelial nitric oxide release with the 2,3-diaminonapthalene assay. AB - The reliable measurement of nitric oxide (NO) production by endothelial cells in vitro has become an important tool for investigating mechanisms of endothelial dysfunction. This study evaluates measuring NO production by cultured porcine pulmonary artery endothelial cells (PAEC) using the assay based on the fluorometric detection of 1-(H)-naphthotriazole, the fluorescent product of the reaction between nitrite (NO2-) and 2,3-diaminonapthalene (DAN). To stimulate NO production, PAEC were treated for 60 min with agonists known to stimulate endothelial NO production. The DAN assay was unable to detect NO production from agonist-stimulated PAEC. In contrast, chemiluminescence analysis, which detects NO, NO2-, and nitrate (NO3-) (collectively referred to as NO(x)), detected significant increases in NO(x) from stimulated PAEC. Nitrate reductase-mediated reduction of NO3-to NO2- in media from stimulated PAEC enhanced the ability of the DAN assay to detect NO release from PAEC. These results provide the first direct comparison of the sensitivity of these two commonly employed assays. Our findings emphasize that NO3-reduction may be required to enable the DAN assay to detect small amounts of NO produced by cultured endothelial cells. PMID- 12654475 TI - Recombinant peroxiredoxin 5 protects against excitotoxic brain lesions in newborn mice. AB - The pathophysiology of brain lesions associated with cerebral palsy is multifactorial and likely involves excess release of glutamate and excess production of free radicals, among other factors. Theoretically, antioxidants could limit the severity of these brain lesions. Peroxiredoxins are a family of peroxidases widely distributed in eukaryotes and prokaryotes. Peroxiredoxin 5 (PRDX5) is a recently discovered mammalian member of this family of antioxidant enzymes that is able to reduce hydrogen peroxide and alkyl hydroperoxides. The present study was designed to examine the neuroprotective effects of recombinant PRDX5 against neonatal excitotoxic challenge in both in vivo and in vitro experiments. For in vivo experiments, mice (postnatal day 5) were injected intraneopallially with ibotenate acting on NMDA and metabotropic receptors, or S bromowillardiine acting on AMPA-kainate receptors to produce excitotoxic stress and brain lesions. Systemically administered recombinant PRDX5 provided protection against ibotenate-induced excitotoxic stress. Brain lesions of animals given ibotenate and PRDX5 were up to 63% smaller than that given ibotenate alone. However, PRDX5 provided no prevention from lesions induced with S bromowillardiine. A mutated recombinant PRDX5 that is devoid of peroxidase activity was also tested and showed no protection against lesions induced by either ibotenate or S-bromowillardiine. Two classical antioxidants, N acetylcysteine and catalase-PEG, provided the same neuroprotective effect as PRDX5. For in vitro experiments, neocortical neurons were exposed to 300 microM NMDA alone, NMDA plus recombinant PRDX5, or NMDA, recombinant PRDX5 and dithiothreitol, a classical electron donor for peroxiredoxins. Recombinant PRDX5 plus dithiothreitol displayed a synergistic neuroprotective effect on NMDA induced neuronal death. These findings indicate that reactive oxygen species production participates in the formation of NMDA receptor-mediated brain lesions in newborn mice and that antioxidant compounds, such as PRDX5, provide some neuroprotection in these models. PMID- 12654476 TI - Effect of SOD1 overexpression on age- and noise-related hearing loss. AB - Reactive oxygen species (ROS) have been implicated in hearing loss associated with aging and noise exposure. Superoxide dismutases (SODs) form a first line of defense against damage mediated by the superoxide anion, the most common ROS. Absence of Cu/Zn SOD (SOD1) has been shown to potentiate hearing loss related to noise exposure and age. Conversely, overexpression of SOD1 may be hypothesized to afford a protection from age- and noise-related hearing loss. This hypothesis may be tested using a transgenic mouse model carrying the human SOD1 gene. Contrary to expectations, here, we report that no protection against age-related hearing loss was observed in mice up to 7 months of age or from noise-induced hearing loss when 8 week old mice were exposed to broadband noise (4-45 kHz, 110 dB for 1 h). Mitochondrial DNA deletion, an index of aging, was elevated in the acoustic nerve of transgenic mice compared to nontransgenic littermates. The results indicate the complexity of oxidative metabolism in the cochlea is greater than previously hypothesized. PMID- 12654477 TI - Vitamin C protects low-density lipoprotein from homocysteine-mediated oxidation. AB - Homocysteine, an atherogenic amino acid, promotes iron-dependent oxidation of low density lipoprotein (LDL). We investigated whether vitamin C, a physiological antioxidant, could protect LDL from homocysteine-mediated oxidation. LDL (0.2 mg of protein/ml) was incubated at 37 degrees C with homocysteine (1000 microM) and ferric iron (10-100 microM) in either the absence (control) or presence of vitamin C (5-250 microM). Under these conditions, vitamin C protected LDL from oxidation as evidenced by an increased lag time preceding lipid diene formation (> or = 5 vs. 2.5 h for control), decreased thiobarbituric acid-reactive substances accumulation (< or = 19 +/- 1 nmol/mg when vitamin C > or = 10 microM vs. 32 +/- 3 nmol/mg for control, p <.01), and decreased lipoprotein anodic electrophoretic mobility. Near-maximal protection was observed at vitamin C concentrations similar to those in human blood (50-100 microM); also, some protection was observed even at low concentrations (5-10 microM). This effect resulted neither from altered iron redox chemistry nor enhanced recycling of vitamin E in LDL. Instead, similar to previous reports for copper-dependent LDL oxidation, we found that vitamin C protected LDL from homocysteine-mediated oxidation through covalent lipoprotein modification involving dehydroascorbic acid. Protection of LDL from homocysteine-mediated oxidation by vitamin C may have implications for the prevention of cardiovascular disease. PMID- 12654478 TI - Pulmonary arterial endothelial cells affect the redox status of coenzyme Q0. AB - The pulmonary endothelium is capable of reducing certain redox-active compounds as they pass from the systemic venous to the arterial circulation. This may have important consequences with regard to the pulmonary and systemic disposition and biochemistry of these compounds. Because quinones comprise an important class of redox-active compounds with a range of physiological, toxicological, and pharmacological activities, the objective of the present study was to determine the fate of a model quinone, coenzyme Q0 (Q), added to the extracellular medium surrounding pulmonary arterial endothelial cells in culture, with particular attention to the effect of the cells on the redox status of Q in the medium. Spectrophotometry, electron paramagnetic resonance (EPR), and high-performance liquid chromatography (HPLC) demonstrated that, when the oxidized form Q is added to the medium surrounding the cells, it is rapidly converted to its quinol form (QH2) with a small concentration of semiquinone (Q*-) also detectable. The isolation of cell plasma membrane proteins revealed an NADH-Q oxidoreductase located on the outer plasma membrane surface, which apparently participates in the reduction process. In addition, once formed the QH2 undergoes a cyanide sensitive oxidation by the cells. Thus, the actual rate of Q reduction by the cells is greater than the net QH2 output from the cells. PMID- 12654479 TI - Reversal of the superoxide dismutase reaction revisited. AB - Reversal of the superoxide dismutase (SOD) reaction was measured in terms of the reduction of tetranitromethane (TNM) by O2-. Cu,ZnSOD caused a biphasic reduction of TNM by H2O2. The rapid initial phase was stoichiometric with the enzyme and was followed by a slower catalytic phase that was oxygen dependent and was augmented by HCO3-. The reaction scheme explaining this behavior is presented and a rate constant for the reduction of O2 by the cuprous enzyme is estimated. This rate constant is so low that it precludes significant O2- production by the reduced enzyme under the conditions explored. PMID- 12654480 TI - Cardiopulmonary bypass as a cause of free radical-induced oxidative stress and enhanced blood-borne isoprostanes in humans. AB - Free radicals are believed to be involved in postsurgery-related complications. We studied whether cardiopulmonary bypass (CPB) operation has any immediate impact on the initiation of oxidative stress and inflammatory response by measuring isoprostanes and prostaglandin F2alpha during and 24 h following CPB. The levels of 8-iso-PGF2alpha (a major F2-isoprostane and biomarker of oxidative stress) and 15-keto-dihydro-PGF2alpha (a major metabolite of PGF2alpha and biomarker of inflammatory response) were measured in frequently collected plasma samples before, during, and up to 24 h postsurgery in 21 patients. 8-Iso PGF2alpha levels significantly increased within 3 min (p <.0001) and continued until 50 min (p <.0001) during CPB. On the contrary, no significant increase of inflammatory response indicator, 15-keto-dihydro-PGF2alpha was found during and up to 24 h postoperatively. These findings establish an increased free radical induced oxidative stress activity rather than inflammatory response after CPB. PMID- 12654482 TI - Inhibition of influenza infection by glutathione. AB - Infection by RNA virus induces oxidative stress in host cells. Accumulating evidence suggests that cellular redox status plays an important role in regulating viral replication and infectivity. In this study, experiments were performed to determine whether the thiol antioxidant glutathione (GSH) blocked influenza viral infection in cultures of Madin-Darby canine kidney cells or human small airway epithelial cells. Protection against production of active virus particles was observed at a low (0.05-0.1) multiplicity of infection (MOI). GSH inhibited expression of viral matrix protein and inhibited virally induced caspase activation and Fas upregulation. In BALB/c mice, inclusion of GSH in the drinking water decreased viral titer in both lung and trachea homogenates 4 d after intranasal inoculation with a mouse-adapted influenza strain A/X-31. Together, the data suggest that the thiol antioxidant GSH has an anti-influenza activity in vitro and in vivo. Oxidative stress or other conditions that deplete GSH in the epithelium of the oral, nasal, and upper airway may, therefore, enhance susceptibility to influenza infection. PMID- 12654481 TI - Impacts of glutathione peroxidase-1 knockout on the protection by injected selenium against the pro-oxidant-induced liver aponecrosis and signaling in selenium-deficient mice. AB - Previous research has suggested that repletion of cellular glutathione peroxidase (GPX1) activity by a single injection of Se was dissociated from the Se protection against the pro-oxidant-induced liver necrosis in Se-deficient rodents. Using the GPX1 knockout (GPX1-/-) mice, TUNEL assay, and apoptosis gene expression microarray, we have demonstrated strikingly different impacts of GPX1 knockout on hepatotoxicity and the related signaling induced by an intraperitoneal injection of 12.5 mg paraquat/kg body weight (b.wt.). In both Se deficient GPX1-/- and wild-type (WT) mice, the paraquat did not induce typical liver necrosis, rather aponecrosis or necrapoptosis, a syncretic process of cell death sharing characteristics of both apoptosis and necrosis. The severity of liver aponecrosis and the associated mortality were reduced to a much greater extent by an injection of Se (ip, 50 microg/kg b.wt. as Na2SeO3) prior to paraquat stress in the WT mice, compared with the GPX1-/- mice. The induced liver aponecrosis seemed to be more apoptotic in the GPX1-/- mice but more necrotic in the WT mice. The paraquat-mediated gene or protein expression of proapoptotic Bax, Bcl-w, and Bcl-X(S), cell survival/death factors GADD45, MDM2, c-Myc, and caspase-3 was upregulated, but that of antiapoptotic Bcl-2 was downregulated in the GPX1-/- mice vs. the WT mice. Overall, these differences between the two groups of mice were related to a low level of liver GPX1 activity in the WT mice that represented < 4% of the normal physiological level. Therefore, the low level of GPX1 activity in the Se-deficient mice can exert a potent role in defending against liver aponecrosis induced by moderate oxidative stress. PMID- 12654484 TI - Molecular based methods for the detection of microbial pathogens in the environment. PMID- 12654483 TI - Activation of NAD(P)H oxidase by lipid hydroperoxides: mechanism of oxidant mediated smooth muscle cytotoxicity. AB - Oxidized lipids, such as 13-hydroperoxyoctadecadienoic acid (13-HPODE), have been implicated in the pathogenesis of atherosclerosis. 13-HPODE, a constituent of oxidized low-density lipoproteins, can induce cytotoxicity of vascular smooth muscle cells (SMC), which may facilitate plaque destabilization and/or rupture. 13-HPODE-induced cytotoxicity has been linked to oxidative stress, although the mechanisms by which this occurs are unknown. In the present study, we show that 13-HPODE and 9-HPODE (10-30 microM) increased superoxide (O2*-) production and induced cytotoxicity in SMC. The 13-HPODE-induced increase in O2*- was blocked by transfecting the cells with antisense oligonucleotides against p22phox, suggesting that the O2*- was produced by NAD(P)H oxidase. Similar concentrations of the corresponding HPODE reduction products, 13-hydroxyoctadecadienoic acid (13 HODE) and 9-HODE, neither increased O2*- production nor induced cytotoxicity, while 4-hydroxy nonenal (4-HNE), an unsaturated aldehyde lipid peroxidation product, induced cytotoxicity without increasing O2*- production. Treatment with superoxide dismutase or Tiron to scavenge O2*-, or transfection with p22phox antisense oligonucleotides to inhibit O2*- production, attenuated 13-HPODE induced cytotoxicity, but not that induced by 4-HNE. These findings suggest that activation of NAD(P)H oxidase, and production of O2*-, play an important role in lipid hydroperoxide-induced smooth muscle cytotoxicity. PMID- 12654485 TI - Application of the real-time PCR for the detection of airborne microbial pathogens in reference to the anthrax spores. AB - To establish the rapid detection method of airborne bacterial spores, we examined Bacillus anthracis spores by real-time PCR. One hundred liters of air were trapped on a filter of an air monitor device. After it was suspended in PBS, spores of B. anthracis were artificially added. The suspension was also heated at 95 degrees C for 15 min and used for real-time PCR using anthrax-specific primers. A single cell of B. anthracis was detected by real-time PCR within 1 h. Our results provide evidence that anthrax spores from the atmosphere can be detected rapidly, suggesting that real-time PCR provides a flexible and powerful tool to prevent epidemics. PMID- 12654486 TI - Detection of pathogenic Vibrio parahaemolyticus in oyster enrichments by real time PCR. AB - A real time polymerase chain reaction (PCR) assay was developed and evaluated to detect the presence of the thermostable direct hemolysin gene (tdh), a current marker of pathogenicity in Vibrio parahaemolyticus. The real time PCR fluorogenic probe and primer set was tested against a panel of numerous strains from 13 different bacterial species. Only V. parahaemolyticus strains possessing the tdh gene generated a fluorescent signal, and no cross-reaction was observed with tdh negative Vibrio or non-Vibrio spp. The assay detected a single colony forming unit (CFU) per reaction of a pure culture template. This sensitivity was achieved when the same template amount per reaction was tested in the presence of 2.5 microl of a tdh negative oyster:APW enrichment (oyster homogenate enriched in alkaline peptone water overnight at 35 degrees C). This real time technique was used to test 131 oyster:APW enrichments from an environmental survey of Alabama oysters collected between March 1999 and September 2000. The results were compared to those previously obtained using a streak plate procedure for culture isolation from the oyster:APW enrichment combined with use of a non-radioactive DNA probe for detection of the tdh gene. Real time PCR detected tdh in 61 samples, whereas the streak plate/probe method detected tdh in 15 samples. Only 24 h was required for detection of pathogenic V. parahaemolyticus in oyster:APW enrichments by real time PCR, whereas the streak plate/probe method required 3 days and was more resource intensive. This study demonstrated that real time PCR is a rapid and reliable technique for detecting V. parahaemolyticus possessing the tdh gene in pure cultures and in oyster enrichments. PMID- 12654487 TI - PCR-based detection of non-indigenous microorganisms in 'pristine' environments. AB - PCR-based technologies are widely employed for the detection of specific microorganisms, and may be applied to the identification of non-indigenous microorganisms in 'pristine' environments. For 'pristine' environments such as those found on the Antarctic continent, the application of these methods to the assessment of environmental contamination from human activities must be treated with caution. Issues such as the possibility of non-human dispersal of organisms, stability and survival of non-indigenous organisms in vivo, the sensitivity, reproducibility and specificity of the PCR process (and particularly primer design) and the sampling regime employed must all be considered in detail. We conclude that despite these limitations, PCR and related technologies offer enormous scope for assessment of both natural and non-indigenous microbial distributions. PMID- 12654488 TI - The use of NASBA for the detection of microbial pathogens in food and environmental samples. AB - The isothermal amplification method nucleic acid sequence-based amplification (NASBA), which amplifies RNA, has been reported as useful for the detection of microbial pathogens in food and environmental samples. Methods have been published for Campylobacter spp., Listeria monocytogenes and Salmonella enterica ser. Enteritidis in various foods and for Cryptosporidium parvum in water. Both 16S rRNA and various mRNAs have been used as target molecules for detection; the latter may have advantages in allowing specific detection of viable cells. Most of the methods to detect pathogens in foods have employed enrichment in nutrient medium prior to NASBA, as this can ensure sensitivity of detection and encourage the detection of only viable target cells. Although a relatively recent method, NASBA has the potential for adoption as a diagnostic tool for environmental pathogens. PMID- 12654489 TI - Molecular methods for the assessment of bacterial viability. AB - A significant number of pathogenic microorganisms can be found in environmental reservoirs (air, water, soil). It is important to assess the viability status of these organisms to determine whether they pose a threat to public health. Classical methods for determining viability are time consuming. Hence, molecular methods have been developed to address this problem. Molecular methods offer speed, sensitivity and specificity. Both DNA and RNA have been analysed using molecular amplification methods such as polymerase chain reaction (PCR), reverse transcriptase PCR (RT-PCR) and nucleic acid sequence-based amplification (NASBA). However, due to the variable persistence of nucleic acids in cells post-death, the correlation between presence of DNA and RNA and viability is not clear-cut. Similarly, the choice of target and sensitivity of the method can significantly affect the validity of the viability assay. This review assesses the molecular methods currently available and evaluates their ability to assess cell viability with emphasis on environmental pathogens. PMID- 12654490 TI - Towards a unified system for detecting waterborne pathogens. AB - Currently, there is no single method to collect, process, and analyze a water sample for all pathogenic microorganisms of interest. Some of the difficulties in developing a universal method include the physical differences between the major pathogen groups (viruses, bacteria, protozoa), efficiently concentrating large volume water samples to detect low target concentrations of certain pathogen groups, removing co-concentrated inhibitors from the sample, and standardizing a culture-independent endpoint detection method. Integrating the disparate technologies into a single, universal, simple method and detection system would represent a significant advance in public health and microbiological water quality analysis. Recent advances in sample collection, on-line sample processing and purification, and DNA microarray technologies may form the basis of a universal method to detect known and emerging waterborne pathogens. This review discusses some of the challenges in developing a universal pathogen detection method, current technology that may be employed to overcome these challenges, and the remaining needs for developing an integrated pathogen detection and monitoring system for source or finished water. PMID- 12654491 TI - Detection of pathogenic bacteria in shellfish using multiplex PCR followed by CovaLink NH microwell plate sandwich hybridization. AB - Outbreak of diseases associated with consumption of raw shellfish especially oysters is a major concern to the seafood industry and public health agencies. A multiplex PCR amplification of targeted gene segments followed by DNA-DNA sandwich hybridization was optimized to detect the etiologic agents. First, a multiplex PCR amplification of hns, spvB, vvh, ctx and tl was developed enabling simultaneous detection of total Salmonella enterica serotype Typhimurium, Vibrio vulnificus, Vibrio cholerae and Vibrio parahaemolyticus from both pure cultures and seeded oysters. Amplicons were then subjected to a colorimetric CovaLink NH microwell plate sandwich hybridization using phosphorylated and biotinlylated oligonucleotide probes, the nucleotide sequences of which were located internal to the amplified DNA. The results from the hybridization with the multiplexed PCR amplified DNA exhibited a high signal/noise ratio ranging between 14.1 and 43.2 measured at 405 nm wavelength. The sensitivity of detection for each pathogen was 10(2) cells/g of oyster tissue homogenate. The results from this study showed that the combination of the multiplex PCR with a colorimetric microwell plate sandwich hybridization assay permits a specific, sensitive, and reproducible system for the detection of the microbial pathogens in shellfish, thereby improving the microbiological safety of shellfish to consumers. PMID- 12654492 TI - Analysis of environmental microbial communities by reverse sample genome probing. AB - Development of fast and accurate methods for monitoring environmental microbial diversity is one of the great challenges in microbiology today. Oligonucleotide probes based on 16S rRNA sequences are widely used to identify bacteria in the environment. However, the successful development of a chip of immobilized 16S rRNA probes for identification of large numbers of species in a single hybridization step has not yet been reported. In reverse sample genome probing (RSGP), labelled total community DNA is hybridized to arrays in which genomes of cultured microorganisms are spotted on a solid support in denatured form. This method has provided useful information on changes in composition of the cultured component of microbial communities in oil fields, the soil rhizhosphere, hydrocarbon-contaminated soils and acid mine drainage sites. Applications and limitations of the method, as well as the prospects of extending RSGP to cover also the as yet uncultured component of microbial communities, are evaluated. PMID- 12654493 TI - New biochip technology for label-free detection of pathogens and their toxins. AB - microSERS is a new biochip technology that uses surface-enhanced Raman scattering (SERS) microscopy for label-free transduction. The biochip itself comprises pixels of capture biomolecules immobilized on a SERS-active metal surface. Once the biochip has been exposed to the sample and the capture biomolecules have selectively bound their ligands, a Raman microscope is used to collect SERS fingerprints from the pixels on the chip. SERS, like other whole-organism fingerprinting techniques, is very specific. Our initial studies have shown that the Gram-positive Listeria and Gram-negative Legionella bacteria, Bacillus spores and Cryptosporidium oocysts can often be identified at the subspecies/strain level on the basis of SERS fingerprints collected from single organisms. Therefore, pathogens can be individually identified by microSERS, even when organisms that cross-react with the capture biomolecules are present in a sample. Moreover, the SERS fingerprint reflects the physiological state of a bacterial cell, e.g., when pathogenic Listeria and Legionella were cultured under conditions known to affect virulence, their SERS fingerprints changed significantly. Similarly, nonviable (e.g., heat- or UV-killed) microorganisms could be differentiated from their viable counterparts by SERS fingerprinting. Finally, microSERS is also capable of the sensitive and highly specific detection of toxins. Toxins that comprised as little as 0.02% by weight of the biomolecule toxin complex produced strong, unique fingerprints when spectra collected from the complexes were subtracted from the spectra of the uncomplexed biomolecules. For example, aflatoxins B(1) and G(1) could be detected and individually identified when biochips bearing pixels of antibody or enzyme capture biomolecules were incubated in samples containing one or both aflatoxins, and the spectra were then collected for 20 s from an area of the biomolecule pixel approximately 1 microm in diameter. In the future, we plan to investigate the use of hyperspectral imaging Raman microscopy for collecting fingerprints from all the pixels on the biochip, individually yet simultaneously, to enable the rapid detection of diverse pathogens and their toxins in a sample, using a single biochip. PMID- 12654494 TI - Detection of bacterial pathogens in environmental samples using DNA microarrays. AB - Polymerase chain reaction (PCR) is an important tool for pathogen detection, but historically, it has not been possible to accurately identify PCR products without sequencing, Southern blots, or dot-blots. Microarrays can be coupled with PCR where they serve as a set of parallel dot-blots to enhance product detection and identification. Microarrays are composed of many discretely located probes on a solid substrate such as glass. Each probe is composed of a sequence that is complimentary to a pathogen-specific gene sequence. PCR is used to amplify one or more genes and the products are then hybridized to the array to identify species specific polymorphism within one or more genes. We illustrate this type of array using 16S rDNA probes suitable for distinguishing between several salmonid pathogens. We also describe the use of microarrays for direct detection of either RNA or DNA without the aid of PCR, although the sensitivity of these systems currently limits their application for pathogen detection. Finally, microarrays can also be used to "fingerprint" bacterial isolates and they can be used to identify diagnostic markers suitable for developing new PCR-based detection assays. We illustrate this type of array for subtyping an important food-borne pathogen, Listeria monocytogenes. PMID- 12654495 TI - Quantitative, multiplexed detection of bacterial pathogens: DNA and protein applications of the Luminex LabMAP system. AB - Escherichia coli, Salmonella, Listeria monocytogenes and Campylobacter jejuni are bacterial pathogens commonly implicated in foodborne illnesses. Generally used detection methods (i.e., culture, biochemical testing, ELISA and nucleic acid amplification) can be laborious, time-consuming and require multiple tests to detect all of the pathogens. Our objective was to develop rapid assays to simultaneously detect these four organisms through the presence of antigen or DNA using the Luminex LabMAP system. For nucleic acid detection, organism-specific capture probes corresponding to the 23S ribosomal RNA gene (rrl) were coupled covalently to LabMAP microspheres. Target molecules included synthetic complementary oligonucleotides and genomic DNA isolated from ATCC type strains or other well-characterized strains of each organism. Universal PCR primers were designed to amplify variable regions of bacterial 23S ribosomal DNA, yielding biotinylated amplicons of 86 to 109 bp in length. Varying quantities of targets were hybridized to the combined microsphere sets, labeled with streptavidin-R phycoerythrin and analyzed on the Luminex(100) system. Results of nucleic acid detection assays, obtained in 30 to 40 min following amplification, correctly and specifically identified each bacterial species with a detection sensitivity of 10(3) to 10(5) genome copies. Capture-sandwich immunoassays were developed with organism-specific antibodies coupled to different microsphere sets. Microspheres were incubated with organism-specific standards and reactivity was assessed with biotinylated detection antibodies and streptavidin-R-phycoerythrin. In the immunoassays, microsphere-associated fluorescence was organism concentration dependent with detectable response at < or = 1000 organisms/ml and with no apparent cross-reactivity. We have demonstrated that the Luminex LabMAP system is a rapid, flexible platform capable of simultaneous, sensitive and specific detection of pathogens. The practical significance of this multiplexing approach would be to provide more timely, economical and comprehensive information than is available with conventional isolation and identification methodologies. PMID- 12654496 TI - Phage display for detection of biological threat agents. AB - The essential element of any immuno-based detector device is the probe that binds analyte and, as a part of the analytical platform, generates a measurable signal. The present review summarizes the state of the art in development of the probes for detection of the biological threat agents: toxins, bacteria, spores and viruses. Traditionally, the probes are antibodies, which are isolated from sera of immunized animals or culture media of hybridomas. However, the "natural" antibodies may have limited application in the new generation of real-time field detectors and monitoring systems, where stress-resistant and inexpensive long livers are required. Phage display is a newcomer in the detection area, whose expertise is development of molecular probes for targeting of various biological structures. The probes can be selection from about billion clone libraries of recombinant phages expressing on their surface a vast variety of peptides and proteins, including antigen-binding fragments of antibodies. The selection procedure, like kind of affinity chromatography, allows separating of phage binders, which are propagated in Escherichia coli bacterial cells and purified using inexpensive technology. Although phage display traditionally is focused more on development of medical preparations and studying molecular recognition in biological systems, there are some examples of its successful use for detection, which are presented in the review. To be used as probes for detection, peptides and antibodies identified by phage display are usually chemically synthesized or produced in bacteria. Another interesting aspect is using of the selected phage itself as a probe in detector devices, like sort of substitute antibodies. This idea is illustrated in the review by "detection" of beta-galactosidase from E. coli with "landscape" phage displaying a dense array of peptide binders on the surface. PMID- 12654497 TI - Discovery of phage display peptide ligands for species-specific detection of Bacillus spores. AB - Short peptides are capable of tight and specific binding to physiological or fortuitous receptors on the surface of cells. These peptides can be used to tag or capture target cells in an assortment of detector platforms. As part of an effort to identify small-molecule ligands for advanced detectors for spores of Bacillus anthracis, the causative agent of anthrax, we are screening (or biopanning) commercial phage display peptide libraries for peptides that bind tightly and selectively to spores of several Bacillus species. In addition to B. anthracis, these species include B. cereus, B. subtilis, and B. globigii. This review summarizes the methods used in our studies, the results from the biopanning experiments, and the characterization of the spore-binding peptides identified to date. Briefly, several unique families of peptides, with consensus sequences< or = seven-amino-acids long, were identified that exhibit preferential binding to spores (but not vegetative cells) of either one or only a few Bacillus species. At least one peptide family binds well to spores of multiple strains of B. anthracis, while binding poorly or not at all to spores of phylogenetically similar species. This review also discusses other points of interest regarding the use of peptide ligands for spore detection and for the detection of other types of cells. PMID- 12654498 TI - Specific and selective biosensor for Salmonella and its detection in the environment. AB - The specific and selective detection of Salmonella typhymurium based on the use of a polyclonal antibody immobilized by the Langmuir-Blodgett method on the surface of a quartz crystal acoustic wave device was demonstrated in liquid samples. These biosensors were selective to S. typhymurium in the presence of large concentrations of Escherichia coli O157:H7. They were also specific to S. typhymurium since bacteria preincubated with free antibody produced no signal. Dark-field and electron microscopy showed that two different antibodies, polyvalent somatic O and flagellar H7, were immobilized on the sensor surface producing two distinct attachments of bacteria at the liquid-solid interface. The somatic O antibody exhibits a rigid, binding, while the flagellar H7 antibody forms a flexible connection allowing a large degree of freedom. When the attachment of bacteria was rigid and strong, the responses of the acoustic wave sensors correlated with changes in the mass of bacteria present at the liquid solid interface. In contrast, when attachment was flexible, the sensor signals were inversely proportional to the additional mass of bound bacteria. This difference is probably determined by the interfacial viscoelasticity and by acoustic and electromagnetic coupling. The signals of environmentally aged sensors with either predominantly rigid or flexible positioning of bacteria were correlated with changes in mass at the liquid-solid interface. Sensors with O or H type of binding could be used for analytical purposes. PMID- 12654499 TI - Expression of the transcriptional coactivators CBP and p300 in the hamster suprachiasmatic nucleus: possible molecular components of the mammalian circadian clock. AB - Immediate early genes are expressed in the mammalian suprachiasmatic nucleus in response to photic information arriving from the retina at restricted times of the day, therefore their expression is regulated by the circadian biological clock. These light-induced genes are also activated by the phosphorylated form of CREB (pCREB) that binds to a cAMP response element upstream of the genes. The nuclear proteins CBP and p300 are known to be coactivators with pCREB in certain cell types, but their identification within the rodent SCN has not been reported. Therefore, in this study we examined the distribution of both CBP and p300 in the hamster suprachiasmatic nucleus. CBP and p300 immunoreactivity is detected in cells throughout the suprachiasmatic nucleus, and the pattern of staining within cells is indicative of a nuclear location for these proteins. The number of cells immunoreactive for both CBP and p300 significantly decreases at mid-night circadian times with respect to mid-day circadian times, although the reduction is less than 20%. Neither CBP nor p300 expression is affected by a circadian phase-resetting light pulse given late in the night. The ability of CBP and p300 to interact with pCREB as well as with the clock gene BMAL1 is discussed, and we propose that CBP and p300 may interact with, and link, both clock genes and clock controlled genes in the generation of circadian rhythms in mammals. We further suggest that there will be a general importance for the role of transcriptional coactivators such as CBP and p300 in many of the molecular pathways related to the mammalian circadian clock. PMID- 12654500 TI - Role of the hippocampal CA2 region following postischemic hypothermia in gerbil. AB - To investigate the changes in the principal subunit of N-methyl-D-aspartate (NMDA) receptor 1 (NR1) following the transient ischemia and postischemic hypothermia, in situ hybridization was used in the gerbil hippocampus. One of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors, Glutamate receptor 2 (GluR2) was also investigated to compare with NR1. Even at 1 day, NR1 and GluR2 mRNAs in the CA1 region were reduced following ischemia. Although postischemic hypothermia prevented almost all the neuronal cell death by ischemia and inhibited the reduction of NR1 and GluR2 mRNAs in the CA1 region after 7 days, the downregulation of NR1 mRNA in the CA2 region was observed even at 1 day. This change was specific for NR1 and not for GluR2. These results suggest that the changes in NR1 and GluR2 receptors at the mRNA level would occur in spite of postischemic hypothermia. The phenomenon in the CA2 region may play an important role to rescue neuronal cell death by ischemia. PMID- 12654501 TI - Gene expression changes in single dentate granule neurons after adrenalectomy of rats. AB - Removal of corticosterone by adrenalectomy induces apoptosis 3 days later, in some, but not all, rat dentate granule cells. We hypothesized that individual dentate cells trigger specific gene expression profiles that partly determine their apoptosis susceptibility. RNA was collected from physiologically characterized granule cells at 2 or 3 days after adrenalectomy or sham operation, and linearly amplified. The amplified RNA was hybridized to cDNA clones of: (1) candidate genes earlier identified after adrenalectomy in whole hippocampi with SAGE; and (2) genes encoding growth factors and their receptors. We observed that based on the entire expression profile, cells relatively resistant to apoptosis 3 days after adrenalectomy clustered together with one-third of cells 2 days after adrenalectomy. Within the group of ADX cells, a limited number of transcript ratios were found to correlate-positively or negatively-with a known risk factor for apoptosis, calcium influx. The overall analysis of physiological properties and multiple gene expression in single cells can narrow down the number of critical genes involved in apoptosis identified with large scale gene screening methods and allows a first impression of their role as being a potential risk factor or neuroprotective. PMID- 12654502 TI - Exogenous brain-derived neurotrophic factor prevents postischemic downregulation of [3H]muscimol binding to GABA(A) receptors in the cortical penumbra. AB - We have previously shown that exogenous application of brain-derived neurotrophic factor (BDNF) reduces infarct volume in the cortical ischemic penumbra after experimental focal ischemia [Stroke 31 (2000) 2212-2217]. Since BDNF is known to modulate the expression and function of various neurotransmitter receptors, we addressed the question whether BDNF may act via modification of postischemic ligand binding to excitatory NMDA and AMPA and/or inhibitory GABA(A) receptors, respectively. Transient focal cerebral ischemia was induced in male Wistar rats for 2 h using the suture occlusion technique. A period of 30 min after occlusion of the middle cerebral artery, BDNF (300 microg/kg per hour in vehicle; n=5) or vehicle alone (n=5) was continuously infused intravenously for 3 h. Using quantitative receptor autoradiography, postischemic ligand binding of [(3)H]MK 801, [(3)H]AMPA and [(3)H]muscimol was analyzed in the ischemic core, the ischemic cortical penumbra and corresponding regions of the contralateral hemisphere. Transient focal ischemia caused a significant reduction of [(3)H]muscimol binding to GABA(A) receptors within the ischemic cortical penumbra of placebo-treated rats. This was largely prevented by exogenous application of BDNF. [(3)H]MK-801 and [(3)H]AMPA binding values were also reduced in the cortical penumbra and the corresponding area of the contralateral hemisphere. Our data suggest that the neuroprotective effect of BDNF against ischemic damage in the cortical penumbra may be mediated in part by maintained activity of the inhibitory GABAergic system which likely counteracts glutamate induced excitotoxicity. PMID- 12654503 TI - Regulation of GRK 2 and 6, beta-arrestin-2 and associated proteins in the prefrontal cortex of drug-free and antidepressant drug-treated subjects with major depression. AB - G protein-coupled receptor kinases (GRKs) and beta-arrestin-2 play a crucial role in the regulation of neurotransmitter receptors in brain. In this study, GRK 2, GRK 6, beta-arrestin-2 and associated proteins (Gbeta proteins and protein phosphatase (PP)-2A) were quantitated in parallel (immunodensity with specific antibodies) in brains of depressed subjects (drug-free and antidepressant treated) to investigate the effect of major depression and antidepressant drugs on these receptor regulatory proteins. Specimens of the prefrontal cortex (Brodmann's area 9) were collected from 19 suicide and non-suicide depressed subjects and 13 control subjects. In drug-free (n=9), but not in antidepressant treated (n=10), depressed subjects an increase in the density of membrane associated GRK 2 (30%, n=9, P=0.005) was found compared with that in sex-, age-, and PMD-matched controls. Comparison between drug-free and antidepressant-treated depressed subjects showed that GRK 2 was reduced in membrane (39%, n=10, P=0.008) and cytosolic (44%, n=10, P=0.09) preparations after antidepressant drug treatment. In contrast, membrane-associated GRK 6 (drug-free and antidepressant treated depressed subjects) was found unchanged when compared with that in matched controls. Similarly, the densities of beta-arrestin-2, PP-2A, and Gbeta proteins were not significantly different from those in matched controls. There was a positive correlation between the immunodensities of GRK 2 and beta-arrestin 2 in membrane preparations (r=0.48, n=19, P=0.04), suggesting that both proteins are regulated in a coordinated manner in brains of depressed subjects. The results of this study indicate that major depression is associated with upregulation of brain GRK 2, but not GRK 6, and that antidepressant drug treatment appears to induce downregulation of GRK 2 protein. PMID- 12654504 TI - Inhibition of H-ras as a treatment for experimental brain C6 glioma. AB - The oncogene H-ras plays an important role in tumor growth and maintenance and could serve as a target treatment for brain tumors. In this study, diallyl disulfide (DADS), an inhibitor of H-ras was used to treat experimental brain glioma in a rat model. One hundred and twenty-five Sprague-Dawley rats (175-200 g) were implanted with 2 x 10(5) C6 glioma cells into the intra striatal region of the brain. Animals were treated with DADS (33 micromol) either before or after implantation of tumor cells. Control animals received soybean oil. Treatment outcome was evaluated based on H-ras expression in tumor tissue, animal's neurological status, tumor size, and life span. Application of DADS 7 days before implantation of tumor cells reduced the tumor size (P<0.05), improved neurological status (P<0.05), and increased the animal life span (P<0.05) when compared to the control group (no treatment). The expression of H-ras was significantly (P<0.05) reduced in brain tumor tissue of animals treated with DADS before implantation. Application of DADS after tumor implantation failed to improve clinical status or life span. This study demonstrates that pretreatment with DADS is capable of inhibiting the expression of H-ras in experimental brain C6 glioma which leads to an improved neurological status and an extended life span in the rat. Higher doses of DADS or other more potent inhibitors need to be used after tumor has been implanted. PMID- 12654505 TI - Activation of mitogen-activated protein kinase pathways during the death of PC12 cells is dependent on the state of differentiation. AB - PC12 cells that are differentiated with NGF and cAMP become totally dependent on these factors for their survival, unlike those that are differentiated with NGF alone. We have asked whether the MAP Kinases, ERKs, JNKs and p38s play a role in the cell death induced by withdrawal of trophic factors on NGF- and NGF/cAMP differentiated PC12 cells. By Western-blot analyses with antibodies directed against the activated forms of these kinases, we show that when the trophic factors were withdrawn, ERK phosphorylation was reduced to very low levels within 1 h in both cases. Changes in the other enzymes were observed only in the NGF/cAMP-differentiated cells, in which the JNK phosphorylation increased about 160% by 6 h and that of p38 increased linearly to at least 18-fold throughout the cell death process. The increases in p38 and JNK phosphorylation were implicated in the death of the cells, since the p38 inhibitor PD169316 and the JNK inhibitor SP600125 were protective. These results demonstrate that the state of differentiation of PC12 cells, a model for the differentiation of sympathetic neurons, determines their vulnerability to cell death by modifying the state of phosphorylation and the regulation of specific kinases implicated in signal transduction pathways that are responsible for the survival or the death of these cells. PMID- 12654506 TI - Identification of nuclear factor 1 (NF1) as a transcriptional modulator of rat A(2A) adenosine receptor. AB - By a combination of PCR and DNA walking technique, we isolated a 4.8-kb DNA fragment containing a 4.3 kb 5'-flanking region and a 0.5-kb 5'-untranslated region of the rat A(2A) adenosine receptor (A(2A)-R) gene. Various lengths of the 5'-flanking region of the A(2A)-R gene were inserted into an expression vector and transfected into several different cell lines for promoter analysis. Our results reveal that a consensus NF1 element (designated as A(2A)-R/NF1), located between bases -2846 and -2827 of the A(2A)-R gene, functions as a repressor for A(2A)-R promoters in the rat brain-derived type-2 astrocyte cell line (RBA2), which expresses no A(2A)-R. Electrophoretic gel mobility shift assay (EMSA) revealed that two A(2A)-R/NF1-protein complexes of RBA2 nuclear extract were formed. Supershift experiments using an anti-NF1 antibody suggest that NF1 proteins exist in both A(2A)-R/NF1-protein complexes. Furthermore, mutations in the conserved NF1 binding site of this A(2A)-R/NF1 element disturbed DNA-protein formation. Thus, NF1 proteins appear to mediate this cell line-specific suppression of A(2A)-R promoters in RBA2 cells. The importance of NF1 proteins in regulating A(2A)-R promoters was further confirmed in another cell line (Siha) which expresses no endogenous A(2A)-R. Moreover, addition of the A(2A) R/NF1element upstream of an irrelevant thymidine kinase (TK) promoter suppressed its promoter activity in Siha cells, but not in RBA2 cells. Thus, the NF1 mediated inhibition of the A(2A)-R promoter was promoter- and cell line-specific. In summary, we have defined a distal negative element (A(2A)-R/NF1) that plays a functional role in modulating the expression of A(2A)-R. PMID- 12654507 TI - Elevated D3 dopamine receptor mRNA in dopaminergic and dopaminoceptive regions of the rat brain in response to morphine. AB - As opiates increase dopamine transmission, we measured the effects of morphine on dopamine-related genes using a real-time optic PCR assay that reliably detects small differences in mRNA in discrete brain regions. Tissue from dopaminoceptive and dopaminergic brain regions was collected from rats injected twice daily for 7 days with saline or increasing doses of morphine. Tissues were assayed for D1, D2 and D3 dopamine receptor mRNAs (D1R, D2R and D3R), as well as for mRNAs for tyrosine hydroxylase (TH) and the dopamine transporter (DAT). The neuron associated mRNAs for SNAP-25 and synaptophysin, as well as the glial-associated mRNA for S100-beta and three 'housekeeping' mRNAs, were also measured. As reported previously by others, there was no alteration in D1R mRNA and a 25% decrease in D2R mRNA in the caudate-putamen, 2 h after the final morphine injection. Importantly, in the same RNA extracts, D3R mRNA showed significant increases of 85% in the caudate-putamen and 165% in the ventral midbrain, including the substantia nigra and ventral tegmental area. There were no other significant morphine effects. Mapping of brain regions in saline control rats agreed with previous studies, including showing the presence of low abundance TH mRNA and the absence of DAT mRNA in the caudate-putamen. The finding that chronic, intermittent injections of morphine caused an increase in D3R mRNA extends our understanding of the ability of D3R agonists to reduce the effects of morphine. PMID- 12654508 TI - Association of the C677T and A1298C polymorphisms in the 5,10 methylenetetrahydrofolate reductase gene in patients with migraine risk. AB - Although controversial, diminished activity of 5,10 methylenetetrahydrofolate reductase (MTHFR), a regulatory enzyme of homocysteine metabolism, may predispose to migraine in Turkish people. In a case-control study, we determined the prevalence of two common MTHFR polymorphisms,C677T and A1298C, in 102 migraine patients (23 migraine with aura, 70 migraine without aura and nine with tension type headache) and compared it to that of 136 healthy controls. The frequencies of the T allele of MTHFR677 and the C allele of MTHFR1298 were significantly higher in the total migraine population (33.82%, 33.82%) than in controls (25.38% and 24.26%), respectively. The genotypes T677T and C1298C were the only genotypes significantly associated with migraine (OR=5.702; 95% CI=1.184-27.457; P=0.015) and (OR=8.933; 95% CI=1.953-40.869; P=0.001), respectively). Individuals with migraine with aura with C1298C and C677C/C1298C genotypes were even more profoundly associated with migraine risk than others (OR=14.105; 95% CI=2.417 82.320; P=0.0001) and (OR=10.050; 95% CI=1.580-63.907; P=0.003), respectively. However individuals with migraine without aura with T677T and C1298C genotypes showed the same susceptibility (OR=7.444; 95% CI=1.503-36.863); P=0.005). Patients with C1298C and C677C/C1298C genotypes may also predispose to tension type headache (OR=8.375; 95% CI=0.685-102.458); P=0.049). PMID- 12654510 TI - Regional and cellular distribution of DREAM in adult rat brain consistent with multiple sensory processing roles. AB - We used immunohistochemistry to map the recently discovered EF-hand protein, DREAM (downstream regulatory element antagonist modulator), and compare its expression in rat brain with that of other key neural proteins. DREAM immunoreactivity was most intense in cerebellar granular cortex. That expression pattern matches one reported for Kv4.2, an interaction partner of DREAM in regulation of potassium channels. On the other hand, the regional and cellular expression of DREAM in cerebellum was opposite to that of calbindin D(28k). Other loci for DREAM expression included the hippocampus and retrosplenial granular cortex, which share afferent and efferent connections. A notable trend, however, was the consistent appearance of DREAM at primary sites for sensory processing. These included the optic tract, superior colliculus, olfactory bulb, and several thalamic relay centers such as the anterior dorsal, medial geniculate, dorsolateral geniculate, ventral posteromedial and ventral posterolateral nuclei. Altogether, the results are consistent with multiple functions for DREAM, including a potential role in transfer of sensory information. PMID- 12654509 TI - Heat shock protein 27 delivered via a herpes simplex virus vector can protect neurons of the hippocampus against kainic-acid-induced cell loss. AB - Heat shock proteins are expressed in response to cellular stress and can protect cells from further stress and facilitate recovery. Heat shock protein 27 is of particular interest because it has been implicated in a range of protective roles including protein chaperoning, stabilising elements of the cytoskeleton and as an active inhibitor of apoptosis. In the present study, we have examined the potential of administration of exogenous HSP27 to confer protection against KA induced neuronal cell death in vivo. We aimed to exploit the neurotropic specificity of herpes simplex virus-1 based virus vectors, which have been rendered replication-incompetent, to infect neurons of the hippocampus. The systemic administration of kainic acid, an analogue of glutamate, causes seizures resulting in neuronal damage and is an established animal model of epilepsy. Neuron loss is particularly prominent in the hippocampus and the mode of death is at least partly apoptotic in nature. We show that the overexpression of HSP27 in these neurons can significantly augment their survival following kainic acid administration. In contrast, injection of a control virus expressing beta galactosidase does not confer protection. This is the first time that protection by exogenously expressed HSP27 has been demonstrated in an in vivo model of neuronal cell death. PMID- 12654511 TI - Pivotal role of attractin in cell survival under oxidative stress in the zitter rat brain with genetic spongiform encephalopathy. AB - Accumulation of reactive oxygen species (ROS), which is generated during energy metabolism, is a cause of physiological aging, neuropathogenesis and numerous diseases, such as Parkinson's and Alzheimer's diseases. Zitter rat is an autosomal recessive mutant, characterized by spongiform degeneration and hypomyelination in the brain, and the phenotype has been suggested to be involved in oxidative stress by the accumulation of ROS. To determine the relation between neurodegeneration of the zitter rat and Attractin (Atrn) gene expression, which was identified as a gene responsible for the zitter, we established fibroblast cells from the zitter rat (Fz) and the Wistar tremor control (WTC) rat (Fw), and transduced Fz cells with the Atrn gene (Fz/Atrn). In the Fz/Atrn cells, accumulation of ROS was repressed, and cell survival against oxidative stress was enhanced to the same level as in Fw cells. Interestingly, phosphorylation of ERK was significantly increased in Fz/Atrn cells by H(2)O(2) stimulus, similarly to Fw cells. Furthermore, activation of ERK was confirmed in the brains of WTC and zitter rats by Western blot analysis and immunohistochemistry. These observations suggested that lack of Atrn gene expression induced neurodegeneration by a decrease in active ERK through an intracellular signaling via oxidative stress. PMID- 12654512 TI - Alterations in brain protein kinase C isoforms following developmental exposure to a polychlorinated biphenyl mixture. AB - PCBs have been shown to alter several neurochemical end-points and are implicated in the etiology of some neurological diseases. Recent in vivo studies from our laboratory indicated that developmental exposure to a commercial PCB mixture, Aroclor 1254, caused perturbations in calcium homeostasis and changes in protein kinase C (PKC) activities in rat brain. However, it is not known which molecular substances are targets for PCB-induced developmental neurotoxicity. Since the PKC signaling pathway has been implicated in the modulation of motor behavior as well as learning and memory, and the roles of PKC are subspecies specific, the present study attempted to analyze the effects on selected PKC isozymes in the cerebellum and the hippocampus following developmental exposure (gestational day 6 through postnatal day 21) to a PCB mixture, Aroclor 1254. The results indicated that the developmental exposure to PCBs caused significant hypothyroxinemia and age dependent alterations in the translocation of PKC isozymes; the effects were greatly significant at postnatal day (PND) 14. Immunoblot analysis of PKC-alpha (alpha) from both cerebellum and hippocampus revealed that developmental exposure to Aroclor 1254 caused a significant decrease in cytosolic fraction and an increase in particulate fraction. There was no significant difference between these two brain regions on the level of fractional changes. However, the ratio between the fractions (particulate/cytosol) from cerebellum only was increased in a dose-dependent manner. Analysis of PKC-gamma (gamma) in cerebellum on PND14 showed a decrease in cytosolic fraction in both dose groups and an increase in particulate fraction at high dose (6 mg/kg) only. The ratio between the two fractions was increased in a dose-dependent manner. In the hippocampus, there was a significant decrease in PKC-gamma in cytosolic fraction of the high-dose group and a significant increase in particulate fraction of the low-dose group. But, the ratio between the fractions showed a significant increase (2.6-fold increase in high dose on PND14). Analysis of PKC-epsilon (epsilon) in cerebellum showed a significant decrease in cytosolic fraction at PND14, while particulate PKand an increase in ratio between fractions at 6 mg/kg on PND14. The results from this study indicate that the patterns of subcellular distributions of PKC isoforms following a developmental PCB exposure were PKC isozyme- and developmental stage specific. Considering the significant role of PKC signaling in motor behavior, learning and memory, it is suggested that altered subcellular distribution of PKC isoforms at critical periods of brain development may be a possible mechanism of PCB-induced neurotoxic effects and that PKC-alpha, gamma, and epsilon may be among the target molecules implicated with PCB-induced neurological impairments during developmental exposure. It is believed that this is the first report successfully identifying PKC isoforms responding to PCBs during developmental exposure. PMID- 12654513 TI - Cloning of a C-terminally truncated NK-1 receptor from guinea-pig nervous system. AB - In order to examine the possibility that some actions of substance P may be mediated by a variant of the neurokinin-1 (NK-1) receptor, we isolated and sequenced the cDNA encoding a truncated NK-1 receptor from guinea-pig celiac ganglion and brain mRNA by two-step RT-PCR based on the 3'RACE method. The truncated NK-1 receptor sequence corresponded to a splice variant missing the final exon 5, and encoded a 311-amino acid protein that was truncated just after transmembrane domain 7, in an identical position to a truncated variant of the human NK-1 receptor. Thus, the truncated NK-1 receptor lacked the intracellular C terminus sequence required for the phosphorylation and internalisation of the full-length NK-1 receptor. Using a sensitive one-step semi-quantitative RT-PCR assay, we detected mRNA for both the full length and truncated NK-1 receptors throughout the brain, spinal cord, sensory and autonomic ganglia, and viscera. Truncated NK-1 receptor mRNA was present in lower quantities than mRNA for the full-length NK-1R in all tissues. Highest levels of mRNA for the truncated NK-1 receptor were detected in coeliac ganglion, spinal cord, basal ganglia and hypothalamus. An antiserum to the N-terminus of the NK-1 receptor labelled dendrites of coeliac ganglion neurons that were not labelled with antisera to the C-terminus of the full length NK-1 receptor. These results show that a C terminally truncated variant of the NK-1 receptor is likely to be widespread in central and peripheral nervous tissue. We predict that this receptor will mediate actions of substance P on neurons where immunohistochemical evidence for a full length NK-1 receptor is lacking. PMID- 12654514 TI - Pro-brain-derived neurotrophic factor is decreased in parietal cortex in Alzheimer's disease. AB - Brain-derived neurotrophic factor (BDNF) promotes the function and survival of the major neuronal types affected in Alzheimer disease, such as hippocampal, cortical and basal forebrain cholinergic neurons. We and others have demonstrated a reduction in BDNF mRNA expression in Alzheimer's disease hippocampus and cortex, which may help to explain the selective vulnerability of these neurons. Several studies have also shown decreased BDNF protein in Alzheimer's disease. BDNF protein is synthesized as a precursor, proBDNF, which is cleaved to the mature 14-kDa form. We demonstrate here that BDNF exists as a mixture of proBDNF and mature BDNF in all regions tested of human brain. Using Western blotting, we observe a 40% reduction in proBDNF levels in Alzheimer's disease parietal cortex compared to controls. Thus, decreased BDNF protein measured by ELISA and immunohistochemistry likely represents a mixture of the two BDNF forms, and previously reported decreases in BDNF protein may be due, at least in part, to a significant reduction in proBDNF levels. Although the biological activity of proBDNF is unknown, reduced proBDNF may have functional consequences for the selective neuronal degeneration in Alzheimer's disease brain. PMID- 12654515 TI - VEGF-induced activation of the PI3-K/Akt pathway reduces mutant SOD1-mediated motor neuron cell death. AB - The increased oxidative stress induced by mutant SOD1 is associated with motor neuron degeneration in both human ALS and transgenic mice expressing mutant SOD1. Vascular endothelial growth factor (VEGF) is neurotrophic and also protects from hypoxia-induced neuronal injury. The potential role of VEGF in preventing mutant SOD1-mediated motor neuron cell death was examined using a mouse NSC34 motor neuron-like cell culture system. Infection with adenovirus containing mutant G93A SOD1, but not vector control or wild-type SOD1, increased cellular oxidative stress and motor neuron-like cell death. However, NSC34 cells pretreated with VEGF displayed a dose-dependent resistance to oxidative damage from hydrogen peroxide, TNF-alpha, and mutant G93A-SOD1. VEGF activated both PI3-K and MAPK activities in mouse NSC34 motor neuron-like cells. Pharmacological inhibitors and constitutively active as well as dominant negative mutants of MAPK and PI3-K revealed that the protective effects of VEGF were mediated via the PI3-K activity, and that MAPK activation was not associated with NSC34 cell survival. Furthermore, VEGF-induced downstream Akt activation promoted motor neuron-like NSC34 cell survival in the presence of mutant G93A-SOD1. Thus, VEGF protected mouse NSC34 motor neuron-like cell death from mutant G93A-SOD1 effects via PI3 K/Akt activation. PMID- 12654516 TI - Cellular and subcellular distributions of translation initiation, elongation and release factors in rat hippocampus. AB - Novel protein synthesis in the brain has been suggested to contribute to the formation of synapses and neural circuits during development and the modulation of long-term synaptic plasticity through life. However, cellular and subcellular distribution of neuronal translation machinery and regulator molecules has not yet been extensively characterized in rat brain. In this report, the distribution of translation factors in the developing hippocampus, a region which is highly plastic, was analyzed by immunohistochemistry and Western blotting. Western blot analysis revealed that the hippocampus expresses the factors in all three steps of translation, initiation factors, elongation factors and a release factor. Immunochemical studies of hippocampal slices and culture showed that all translation factors were observed not only in cell bodies but also in dendrites of hippocampal neurons. In addition, the levels of the individual translation factors differed between hippocampal subregions. The differential distribution of translation factors was also confirmed by Western blotting. These results suggest that regulated protein synthesis occurs in the hippocampus, with differences existing between different subregions such as CA1, CA3 and dentate gyrus. PMID- 12654517 TI - How does prolonged caloric restriction ameliorate age-related impairment of long term potentiation in the hippocampus? AB - Prolonged dietary restriction has been reported to suppress age-induced phenomena. In order to investigate how prolonged caloric restriction reduces age related deterioration of hippocampal synaptic transmission, we compared the levels of major hippocampal polyunsaturated fatty acids, arachidonic acid and docosahexaenoic acid between 4- and 26-month-old rats. The Ca(2+) responses upon perfusion of NMDA or 30 mM K(+) between 4- and 26-month-old rats with prolonged dietary restriction were also compared using the fluorescent probe Fura-2. A decrease in membrane arachidonic acid is thought to be a major causal factor in the age-related impairment of long-term potentiation. Long-term caloric restriction seems to increase arachidonic acid levels regardless of age. However, there is no significant difference of hippocampal arachidonic acid levels between in freely feeding 4- and 26-month-old rats. Similar results were obtained from the measurement of hippocampal docosahexaenoic acid levels. Under caloric restriction, the 500 microM N-methyl-D-aspartate-induced Ca(2+) response was greatly reduced by aging, while the 30 mM K(+)-induced Ca(2+) response was not affected. In our preliminary data, the amplitude of the population spike after tetanic stimulation did not differ between 4- and 26-month-old rats under caloric restriction, while 50 microM of 2-amino-5-phosphonovaleric acid, a N-methyl-D aspartate antagonist, markedly inhibited a potentiation of the population spike in 4-month-old rats, but with negligible inhibition in 26-month-old rats. From these results, an age-related impairment of hippocampal excitatory synaptic transmission may not be solely due to the reduction of membrane arachidonic acid. Caloric restriction might prevent age-related reduction in hippocampal synaptic transmission by enhancing non-N-methyl-D-aspartate mechanisms. PMID- 12654518 TI - Dynamic changes in prefrontal cortex gene expression following lysergic acid diethylamide administration. AB - Lysergic acid diethylamide (LSD) is a psychoactive drug that transiently alters human perception, behavior, and mood at extremely low doses. Certain aspects of the behavior elicited by acute doses of LSD closely resemble symptoms of mental disorders such as schizophrenia. Characterizing gene expression profiles after LSD will be important for understanding how it alters behavior, and will lead to novel insights into disorders, such as schizophrenia, whose behavioral symptoms resemble the temporary effects of hallucinogenic drugs. We previously identified a small collection of genes within the rat prefrontal cortex that respond to LSD. Many of the products of these genes are involved in the process of synaptic plasticity. In the current report, we present a detailed analysis of the expression of these genes within the brain using RNase protection analysis. We find that the gene response to LSD is quite dynamic. The expression of some genes increases rapidly and decreases rapidly, while other genes change more gradually. Dose-response studies show two classes of expression; gene expression maximally stimulated at lower doses, versus gene expression that continues to rise at the higher doses. The role of the 5-HT(1A) and 5-HT(2A) receptor in mediating the increases in gene expression was examined in a series of experiments using receptor specific antagonists. Most expression increases were due to activation of the 5-HT(2A) receptor, however expression of two genes had neither a 5-HT(1A) nor a 5-HT(2A) receptor component. PMID- 12654519 TI - Glutamate stimulates neurotrophin expression in cultured Muller cells. AB - The uptake of excess extracellular glutamate and the secretion of neurotrophins by glial cells have been suggested to protect CNS neurons from glutamate-induced toxicity. In the retina, perturbation of glutamate transport and decreased retrograde transport of neurotrophic factors such as brain-derived neurotrophic factor (BDNF) may contribute to ganglion cell death in experimental glaucoma. Although many studies show a clear relationship between glutamate and neurotrophic factors, such relationship has not been thoroughly investigated in the retinal environment. In the following study, we determined the effects of glutamate on early passaged rat Muller cells, specifically their expression of neurotrophic factors including BDNF, nerve growth factor (NGF), neurotrophin-3 (NT-3), neurotrophin-4 (NT-4), and glial-cell line derived neurotrophic factor (GDNF); and of glutamate receptors and transporters using immunoblots or enzyme linked immunosorbent assays. Binding of BDNF to its cognate receptor TrkB was also determined using co-immunoprecipitation studies. Cultured Muller cells grown in the presence of glutamate were also assayed for survival using 3-(4,5 dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS). Our study showed that while glutamate treatment did not promote cell death, it upregulated secretion of BDNF, NGF, NT-3, NT-4, and GDNF by Muller cells. While solitary bands at approximately 13-14 kDa were observed for NGF, NT 3, and NT-4; two BDNF-reactive bands were observed in immunoblots: a faster migrating band at the reported size of the BDNF monomer (approximately 13 kDa); and a more intense band at approximately 36 kDa. GDNF-reactive bands were observed at approximately 22, approximately 28, and approximately 55 kDa. Glutamate also induced significant changes in glutamate receptor and transporter proteins, as well maintained the association of BDNF to TrkB in Muller cells. The decreased N-methyl-D-aspartate receptor (NMDAR) levels and sustained activation of TrkB by BDNF could serve as protective mechanisms for Muller cell survival. Moreover, the increased secretion of neurotrophic factors and upregulation of L glutamate/L-aspartate transporter (GLAST) expression in Muller cells may protect retinal neurons from glutamate toxicity. PMID- 12654520 TI - Molecular cloning and differential expression of the cat immediate early gene c fos. AB - Recently, the effect of binocular central retinal lesions on the expression of immediate early genes in the visual system of adult cats was demonstrated using in situ hybridization and immunocytochemistry. The present study was undertaken to quantify cat c-fos mRNA expression differences in the cat primary visual cortex after sensory deafferentation. Prior to quantification, DNA fragments obtained using reverse transcription-polymerase chain reaction (RT-PCR) in combination with rapid amplification of complementary DNA ends (RACE) were cloned and sequenced. This provided us with the necessary sequence(1) information to prepare cat-specific c-fos primers for the development of a new quantitative RT PCR assay. We optimized a reverse transcription-competitive polymerase chain reaction (RT-cPCR) method with a heterologous DNA fragment (competitor) as external standard to quantify relative amounts of cat c-fos mRNA expression levels. Internal standardization was accomplished by quantifying, in a parallel RT-cPCR, a well-characterized housekeeping gene, glyceraldehyde-3-phosphate dehydrogenase (GAPDH). This cat-specific RT-cPCR assay allowed us to measure c fos mRNA expression levels in central and peripheral regions of primary visual cortex in normal and retinal lesion cats. PMID- 12654522 TI - Cloning of mouse Ca2+/calmodulin-dependent protein kinase kinase beta (CaMKKbeta) and characterization of CaMKKbeta and CaMKKalpha distribution in the adult mouse brain. AB - The Ca(2+)/calmodulin-dependent protein kinase kinases alpha and beta (CaMKKs alpha and beta) are novel members of the CaM kinase family. The CaMKKbeta was cloned from mouse brain. The deduced amino acid sequence shared 96.43% homology with the rat CaMKKbeta. Both the alpha and beta isoforms were widely distributed throughout the adult mouse brain. Additionally, all peripheral tissues examined displayed CaMKK alpha and beta expression. PMID- 12654521 TI - Positive and negative regulators for neuronal BC1 RNA transcription by RNA polymerase III are possible members of the RNA polymerase II transcription system. AB - Neuronal cell-specific BC1 RNA is a unique RNA polymerase III (Pol III) transcript. The transcription is controlled by an activator E2 site and by BCRE, a repressor element, in response to neuronal activity. BC1 RNA is localized to dendritic domains as ribonucleoprotein particles, and it has been suggested to play a functional role in translational regulation of dendritic mRNAs. In the present study, using a luciferase assay in NG108-15 cells, we found that the positive and negative regulators for BC1 RNA transcription can also function in the Pol II transcription system. Our results suggest that the neuronal activity dependent expression of BC1 RNA by Pol III and a subset of neuronal mRNAs by Pol II may be simultaneously controlled by the E2 site and BCRE, as well as their binding proteins. PMID- 12654523 TI - A simple DNA extraction and rapid specific identification technique for single cells and early embryos of two breeds of Bos taurus. AB - In the process of nuclear transfer (NT), different cytoplasm from a donor cell and a recipient oocyte are mixed. However, it is unclear what effect the donor cytoplasm has upon the dedifferentiation of donor nuclei in enucleated ooplasm and upon subsequent production of live cloned offspring. Mitochondria are component parts of cytoplasm so the detection of mitochondrial DNA is helpful to reveal changes of donor cytoplasm in the NT reconstructed embryos. In this study, the experiments were designed to develop efficient DNA extraction techniques and specific primer pairs for mitochondrial DNA of Holstein and Chinese Yellow breeds in order to identify the changes of donor cytoplasm in early stage embryos. Firstly, by adding Triton X-100 and Taq DNA polymerase reaction buffer to the DNA extraction mixture, DNA was rapidly isolated from single diploid cells, single oocytes, early stage embryos and from single hairs. Secondly, two specific primer pairs for the two breeds were designed to detect the cytoplasmic DNA in a different amount of single cells and in early stage embryos. The results show that two specific fragments were successfully amplified from single somatic cells, single oocytes, parthenogenetic embryos and from NT reconstructed embryos. As a result, the techniques provide a powerful tool for studying the developmental mechanism in NT reconstructed embryos. PMID- 12654524 TI - Responses of the estrous cycle in dairy cows exposed to electric and magnetic fields (60 Hz) during 8-h photoperiods. AB - To study the effects of exposure to extremely low frequency (ELF) electric and magnetic fields (EMF) on the estrous cycle of dairy cows under short-day photoperiod, 16 non-lactating, non-pregnant Holstein cows were exposed to a vertical electric field of 10 kV/m and a horizontal magnetic field of 30 microT for 16 h per day in a cross-over design consisting of two sequences. Each sequence included three periods, and each period corresponded to the duration of one estrous cycle. All animals were maintained under short photoperiod (8 h light/16 h dark) during the trial. Exposure to EMF had an impact on the duration of a complete estrous cycle (P<0.01) and on the duration of the luteal phase (P<0.01). The mean duration of one cycle was 19.5+/-0.4 for the control and 21.3+/-0.4 days for the exposed animals, respectively. The mean duration of the luteal phase was 15.4+/-0.4 days for the control and 17.2+/-0.4 days for the exposed group. The total area under the progesterone (P(4)) curve, the amplitude of the curve or the slope of the P(4) rise at the onset of the luteal phase were not affected by EMF exposure. Results indicate that exposure to EMF may increase the duration of the estrous cycle. PMID- 12654525 TI - Influences of epidermal growth factor and insulin-like growth factor-I on bovine blastocyst development in vitro. AB - Experiments were carried out to investigate putative beneficial effects of adding epidermal growth factor (EGF) or insulin-like growth factor-I (IGF-I) for bovine embryo culture in chemically defined media. Presumptive zygotes (18 h post insemination) were randomly assigned to culture treatments. In experiment 1, treatments involved additions of recombinant human EGF to provide concentrations of 0 ng (control), 1, 5, and 25 ng/ml. No differences were seen in numbers of 4 cell stage embryos between groups. A concentration of 5 ng/ml EGF but not 1 or 25 ng/ml during embryo culture improved percentages of 4-cell stage embryos reaching blastocysts compared to the control (P<0.05). Numbers of inner cell mass (ICM) cells and trophoblast cells of day 8 blastocysts were similar for the control and 5 ng/ml EGF-treated groups. In experiment 2, culture with recombinant human IGF-I in concentrations of 0 ng (control), 2, 10, and 50 ng/ml resulted in no differences in numbers of 4-cell stage embryos between groups. When compared to controls, IGF-I treatments at 10 and 50 ng/ml improved proportions of 4-cell stage embryos that reached blastocysts (P<0.05). In experiment 3, numbers of ICM cells of day 8 blastocysts were significantly higher after being cultured with 50 ng/ml of IGF-I compared to those of the controls (P<0.05). No additive effect of combining EGF (5 ng/ml) and IGF-I (50 ng/ml) was seen when results were compared to those following supplementation of the media with either EGF or IGF-I alone. In conclusion, both EGF and IGF-I could independently enhance bovine preimplantational development in chemically defined media and IGF-I but not EGF may play a mitogenic role during early bovine development. PMID- 12654526 TI - Influence of the duration of gamete interaction on cleavage, growth rate and sex distribution of in vitro produced bovine embryos. AB - Various factors including the length of gamete interaction and embryo culture conditions are known to influence the rate of development and sex ratio of mammalian embryos produced in vitro. While the duration of gamete interaction deemed optimum would vary depending upon the species involved and the preferred sex in the outcome of in vitro procedures, the mechanisms favoring the selection of embryos of one sex over the other, or the exact time of post-fertilization stage at which a sex-related difference in growth rate is manifested, are not fully understood. In order to determine the optimum length of gamete co incubation and the impact of male gamete 'aging' on the growth rate and sex ratio of bovine embryos, a series of experiments was carried out using in vitro matured (IVM) oocytes. In experiment 1, IVM oocytes were co-incubated with sperm from two different bulls for 6, 9, 12 and 18 h and the presumptive zygotes were cultured for approximately 7.5 days (178-180 h post-insemination (hpi)) prior to assessing the cleavage rate, blastocyst yield and the sex ratio of blastocysts in each co incubation group. In experiment 2, the blastocysts obtained from different co incubation groups were subjected to differential staining to determine the total cell number (TCN) and the proportion of cells allocated to the inner cell mass (ICM) in male and female embryos to test for sex-related differences in cell proliferation or in differentiation of the two embryonic cell lineages in the blastocysts. In experiment 3, IVM oocytes co-incubated for 6, 9, 12 and 18 h with sperm from a single bull, were cultured for 3 days (72 hpi) and the pre-morulae, categorized according to the specific stage of early development, were sexed to determine if a sex-dependent difference is detectable before the blastocyst stage. In experiment 4, IVM oocytes exposed to prolonged co-incubation (18 and 24 h) were compared with those co-incubated with "aged" (pre-incubated) sperm to determine if "aging sperm" is a factor affecting the growth rate and sex ratio of the out come. Our experiments showed that (1) the shortest period (6 h) allowed the highest proportion of cleaved oocytes to reach the blastocyst stage regardless of the semen donor, (2) males out number females (over 2 to 1) among blastocysts when co-incubation of gametes is reduced to 6 h, (3) the male blastocysts display higher total cell count, and (4) the faster growth rate of the male embryos does not affect the early differentiation and allocation of cells to the ICM. Furthermore, our results indicate that the disruption of the expected 1:1 ratio for male and female embryos in the short term co-incubation group is evident as early as the 4-cell stage and peaks at the 8-cell stage and that prolonged gamete interaction tends to reduce the blastocyst yield to even out the sex ratio. Absence of a significant effect on the yield and sex ratio of blastocysts in the prolonged co-incubation groups irrespective of the type of sperm (aged versus non-aged) used suggest that the preponderance of male embryos in short term gamete interaction group may be dependent upon the in vitro advantage of the Y-chromosome bearing sperm. This advantage, manifested in the precocious development during the pre-morulae stage is confined to a short duration that is neutralized when gamete interaction is allowed to proceed beyond 6h. PMID- 12654527 TI - Effects of pre-synchronization using combinations PGF(2alpha) and (or) GnRH on pregnancy rates of Ovsynch- and Cosynch-treated lactating Holstein cows. AB - In Experiment 1, the effects of two pre-synchronization treatments on synchronized AI pregnancy rates of lactating dairy cattle were compared. Lactating Holstein cows (n=159) received 100 microg of GnRH (im) on day -7 and 25mg of PGF(2alpha) (im) on day 0 and were observed once daily for signs of estrus from day -3 to day 3. Cows detected in standing estrus and those that had lost significant amounts of tail-chalk in the previous 24h were immediately inseminated in a once-daily observation/AI program. Cows not detected in estrus by 72 h after PGF(2alpha) received fixed-time AI (TAI) and a concurrent 100 microg injection of GnRH (im). Cows were randomly assigned by parity and calving date to receive one of the following pre-synchronization treatments: (1) 25mg of PGF(2alpha) (im) on day -35 and day -21 (PGF-PGF) or (2) 100 microg of GnRH (im) on day -14 (GnRH). Fewer (P<0.05) GnRH- (49%, 41/84) than PGF-PGF-pretreated cows (65%, 49/75) were detected in estrus, however, overall pregnancy rates were not affected by pre-synchronization treatment (30 versus 32%, respectively). In Experiment 2, lactating Holstein cows received 100 microg of GnRH (im) on day -7, 25mg of PGF(2alpha) (im) on day 0 and TAI at 60-64 h after PGF(2alpha). Cows were randomized by parity and postpartum interval into pre- and post-synchronization treatments in a 2 x 2 factorial design. Pre-synchronization treatments included: (1) 25mg of PGF(2alpha) (im) on day -35 and on day -21 (PGF-PGF; n=168) or (2) 25mg of PGF(2alpha) (im) on day -21 and 100 microg of GnRH (im) on day -14 (PGF GnRH; n=180). Within each pre-synchronization treatment, cows were further allocated by parity and postpartum interval to receive as a post-synchronization treatment 100 microg of GnRH (im) at either 48 h (Ovsynch; n=175) or 60-64 h (Cosynch; n=173) after PGF(2alpha). Pregnancy rates at TAI were not affected by pre- (PGF-PGF=26%, 44/168 versus PGF-GnRH=24%, 44/180) or post-synchronization treatments (Ovsynch=29%, 50/175 versus Cosynch=22%, 38/173). However, the numeric shift towards reduced pregnancy rates in Cosynch-treated cows suggests the 12h interval between GnRH and AI may be important to optimize conception rates in GnRH-PGF(2alpha)-based TAI protocols in dairy cattle. In conclusion, each of the pre-synchronization protocols evaluated in present study performed with comparable efficacy. Although the Cosynch protocol facilitates more efficient labor utilization, numeric trends toward reduced conception warrants further investigation. PMID- 12654528 TI - Undernutrition of ewe lambs in utero and in early post-natal life does not affect hypothalamic-pituitary function in adulthood. AB - The effect of undernutrition in utero, during late gestation (from day 100), and early neonatal life on hypothalamic-pituitary function was investigated in female lambs born to ewes fed rations calculated to provide either 100% (high; H) or 70% (low; L) of the energy requirements to sustain a twin pregnancy. Following parturition in early spring, ewes and lambs were maintained on pasture with sward heights of 6 cm (H) or 4 cm (L) until week 8 of lactation and then sward heights of 5 cm (H) or 3 cm (L) until weaning at week 14. Mean lamb birth weights were 18% lower in L than H animals (P<0.05) and mean liveweights were 23% lower in the L animals (P<0.001) at weaning at 14 weeks of age. Liveweight differences were not significant at, or after, 26 weeks of age. There were no significant differences between pre-pubertal H and L animals, either before (26 weeks) or after ovariectomy (31 weeks), with respect to hypothalamic or pituitary activity, as measured by LH pulse frequency, pulse amplitude or mean plasma LH and FSH concentrations and the responses to GnRH injection as measured by LH peak amplitude, respectively. Similarly there were no differences in any of these variables in pubertal animals at 18 months of age. At 31 weeks of age, H animals had significantly lower pituitary GnRH receptor binding (P<0.01) and lower ERalpha mRNA content (P<0.05) than L lambs. There were no differences with treatment in the abundance of mRNA for LHbeta, FSHbeta or GnRH-receptor at 31 weeks of age or in pubertal animals aged 18 months, when there were no significant differences with treatment in GnRH receptor binding or ERalpha mRNA expression. It is concluded that effects on lifetime reproductive function of female sheep of undernutrition during late gestation and early neonatal life are unlikely to be expressed through permanent changes in hypothalamic-pituitary function and are therefore attributable to effects exerted directly on the ovary. PMID- 12654530 TI - Germ cell development in Meishan and White Composite gilts. AB - This study compared dynamics of the germ cell population in two swine breeds that differ in prolifacy, White Composite (WC) and Meishan (MS), during fetal and neonatal life and in mature sows. Germ cell populations developed in a similar pattern in these two diverse breeds during fetal life. Maximal germ cell number was observed at 90 days postcoitum (dpc) in both WC and MS gilts, and substantial oogonial apoptosis was evident thereafter with approximately 30% of maximal numbers present at 25 days postpartum (dpp). Neither gilt nor sow germ cell number was correlated with maternal ovulation rate. Postnatal MS gilts had larger pools of primordial follicles and consistently greater proportions and numbers of primary and secondary follicles compared to postnatal WC gilts, indicative of enhanced follicular recruitment and primordial follicle activation. Occasional antral follicles were present in MS ovaries by 25 dpp and numerous surface follicles were observed at 56 dpp in MS but not WC ovaries, indicative of more rapid ovarian maturation and early onset of puberty. Total germ cell number is unlikely to influence or to predict subsequent ovulation rate. These observations highlight important developmental events during late fetal and early postnatal life that prepare the ovarian environment for early onset of puberty and subsequent ovulation in MS gilts. PMID- 12654529 TI - The role of GABA(A) receptors in the neural systems of the medial preoptic area in the control of GnRH release in ewes during follicular phase. AB - To examine the role of gamma-aminobutyric acid (GABA)(A) receptor mediating systems in the control of gonadotropin-releasing hormone (GnRH) release from the medial preoptic area (MPOA) of ewes during the follicular phase of the estrous cycle, the extracellular concentrations of GnRH, beta-endorphin, noradrenaline (NE), dopamine (DA), 4-hydroxy-3-methoxy-phenyl-glycol (MHPG) and 3,4-dihydroxy phenylacetic acid (DOPAC) were quantified during the local infusion of muscimol and bicuculline (agonist and antagonist of GABA(A) receptors, respectively) to this structure. Stimulation of GABA(A) receptors markedly attenuated GnRH release, increased beta-endorphin release and noradrenergic system activity in the MPOA. The decrease of the luteinizing hormone (LH) concentration in blood plasma and LH pulse amplitude suggests that a GABA(A) receptor agonist in the MPOA also suppresses GnRH release from the GnRH axon terminals in the ventromedial hypothalamus/nucleus infundibularis region (VEN/NI). Blockade of GABA(A) receptors had no evident effect on GnRH/LH secretion but decreased beta endorphin release and increased the extracellular DOPAC concentration. The suppressive influence of muscimol in the MPOA on GnRH release might be considered a net result of its direct inhibitory effect on GnRH release, indirect inhibitory influence on GnRH release through activation of the beta-endorphinergic system, and facilitation of GnRH neurons by increasing noradrenaline release. The results obtained during bicuculline perfusion on these systems' activity are not sufficiently consistent to provide a clear understanding of the lack of changes in the GnRH/LH release under blockade of GABA(A) receptors. We conclude that the MPOA in ewes during the follicular phase is an important regulatory site where stimulation of GABA(A) receptors both decreases GnRH secretion and increases beta endorphin release. PMID- 12654531 TI - Cloning and sequence analysis of FSH and LH in the giant panda (Ailuropoda melanoleuca). AB - The giant panda (Ailuropoda melanoleuca) is an endangered species and indigenous to China. It has been proposed that it has a highly specialized reproductive pattern with low fecundity, but little is known about its basic reproductive biology at the molecular level. In this report the genes encoding gonadotropin subunits alpha, follicle-stimulating hormone (FSH) beta and luteinizing hormone (LH) beta of the giant panda were amplified for the first time by RT-PCR from pituitary total RNA, and were cloned, sequenced and analyzed. The results revealed that the open reading region (ORF) of gonadotropin subunits alpha, FSH beta and LH beta are 363, 390 and 426 bp long, respectively. They displayed a reasonably high degree (74-94, 85-93, 75-91%, for alpha, FSH beta and LH beta subunits, respectively) of identity when deduced amino acids were compared with homologous sequences from partial available mammals including human, cattle, sheep, pig, rat, mouse. Three distinct differences were found at the site of 59 aa of the alpha subunit and 55 aa, 68 aa of FSH beta subunit. Our results provide an insight into understanding the mechanism of reproduction regulation and genetic characteristics of giant panda which will make an actual contribution to its conservation. In addition they lay a foundation for a further study towards producing recombinant panda FSH and LH which can be used in artificial breeding aimed to increase its captive reproductive efficiency. PMID- 12654532 TI - The use of steroid hormones in superovulation of Nelore donors at different stages of estrous cycle. AB - The objective of the present study was to evaluate the superovulatory response and ova/embryo recovery from Nelore donors following treatment with a controlled internal drug releasing device and estradiol benzoate (CIDR-B program) at different stages of the estrous cycle. The control group (TI; n=40) received a standard superovulation protocol with females of this group being between days 9 and 12 of the estrous cycle (estrus = day 0). The donors that received a CIDR-B program containing 1.9 g progesterone and an intramuscular injection of estradiol benzoate (2 mg) were at day 0 (TII; n=30), between days 2 and 6 (TIII; n=30), days 7 and 12 (TIV; n=30), days 13 and 16 (TV; n=30) and days 17 and 20 (TVI; n=30) of the estrous cycle. Superovulation was induced with 400 IU of p-FSH, divided into eight decreasing doses (80/80; 60/60; 40/40; 20/20) at intervals of 12h. The donors received PGF2alpha (Cloprostenol) 48 h after beginning the treatment and CIDRs were removed 12h later. Artificial inseminations (AI) were performed 12 and 22 h after the initiation of estrus and embryos were collected 7 days after AI. The mean numbers (+/-S.E.M.) of total ova and embryos, viable (transferable) and degenerated embryos were 14.2+/-11.3, 7.4+/-6.9 and 3.2+/-3.5 (TI), 13.3+/-10.4, 7.1+/-6.2 and 3.3+/-4.3 (TII), 13.5+/-7.0, 8.1+/-6.7 and 2.3+/ 3.0 (TIII), 17.4+/-9.9, 9.4+/-6.9 and 4.0+/-4.4 (TIV), 16.9+/-8.8, 9.8+/-8.1 and 2.7+/-2.5 (TV) and 13.0+/-7.8, 7.2+/-6.9 and 2.3+/-2.5 (TVI), with no significant differences (P>/=0.05) among groups. Pregnancy rates of 67.1% (TI; n=86/128), 60.8% (TII; n=59/97), 62.5% (TIII; n=73/115), 64.1% (TIV; n=84/131), 72.3% (TV; n=81/112) and 60.6% (TVI; n=63/104) were obtained with embryos transferred from these collections and did not differ significantly (P>/=0.05) among groups. The results of the present study allow us to conclude that a combination of steroid hormones may be used prior to superovulation in Nelore donors, at any stage of the estrous cycle without affecting the efficiency of embryo transfer programs. PMID- 12654533 TI - Stop talking at the back. AB - A new compound has been shown to interfere with quroum sensing in Pseudomonas aeruginosa, which could lead to a new class of antibiotics. PMID- 12654534 TI - Centenarians provide genetic clue to age-related disease. PMID- 12654535 TI - Improving drug response with pharmacogenomics. AB - Adverse drug reactions, which in the USA are estimated to account for 100,000 hospitalizations annually, could be halved by the implementation of personalized medicine, but medical schools must adapt to change, say researchers in Israel. PMID- 12654539 TI - Taking the lead? PMID- 12654538 TI - Ancient viruses in the fight against HIV. PMID- 12654540 TI - Inducing apoptosis: mind the natural killer cells. PMID- 12654541 TI - The role of telomeres and telomerase in cancer. PMID- 12654542 TI - Targeting RNA: new opportunities to address drugless targets. AB - Historically, pharmaceutical industries have focussed on the discovery of compounds that target the protein products of genes. The intermediary product between gene and protein, consisting of RNA, has remained largely unexplored. Several drugs targeting the rRNA of bacteria have been, however, in clinical use for over half a century. One of these drug classes, the aminoglycoside antibiotics, also targets human rRNA, and have been developed as therapeutics for genetic disorders. Targeting at the RNA level is an economical approach to address non-drugable proteins and targets that have failed to give leads by hits in HTS, as it can build on biological knowledge gathered over years. RNA also offers entirely new opportunities for drug development, such as targeting of non coding RNA sequences. PMID- 12654543 TI - Potential for proteasome inhibition in the treatment of cancer. AB - Proteasome inhibition is a new approach to treating cancer. Proteasome inhibitors specifically induce apoptosis in cancer cells, but most proteasome inhibitors are not suitable for clinical development. Peptide boronates overcome the shortcomings of earlier generation proteasome inhibitors, and bortezomib (VELCADE; formerly PS-341) is the first peptide boronate to enter clinical trials. Preclinical studies of bortezomib have demonstrated antitumor activity in a variety of tumor types. Phase I trials provided evidence of manageable toxicities and support a twice-weekly dosing regimen now being examined in a Phase III study. PMID- 12654544 TI - Pharmaceutical profiling in drug discovery. AB - Drug discovery research organizations are building capability for measuring an ensemble of crucial 'drug-like' properties. These structure-property relationship (SPR) data complement current SAR information. This pharmaceutical profiling strategy enables research teams to better plan and interpret discovery experiments, be alerted to potential 'show stoppers', improve property liabilities, and select the best candidates for advancement. High throughput property assays for physicochemical properties--solubility, permeability, lipophilicity, stability, and pK(a)--in vitro ADME--metabolism, transporters, protein binding and CYP inhibition--and in vivo PK/exposure provide a wealth of data for teams to make informed decisions. PMID- 12654547 TI - Fracture toughness and microhardness of a composite: do they correlate? AB - OBJECTIVES: Chipping and bulk fracture are major contributors in clinical failures of composite restorations. Fracture toughness (K(Ic)) quantifies susceptibility for fracture, but experimental determination is complicated. It would be beneficial for the dental community if a relatively simple experiment, such as microhardness (HK), could be used to screen composites for fracture resistance. This study explores a possible correlation between K(Ic) and HK. METHODS: Composite cylinders (4mm diameter and approximately 7 mm long) were cured for five combinations of light intensity (I, microm W/cm(2)) and curing time (T, s) to achieve a range of different total light energy densities (I x T=100 x 10, 100 x 20, 300 x 20, 300 x 40, and 700 x 60 microm W s/cm(2)). A chevron-notch was cut in the median plane of the cylinders for the fracture toughness test, which was executed in a displacement control mode at 6 micro m/s cross-head speed (sample size 4). Knoop hardness was determined at the median plane of the cylinders (sample size 6). The tests were performed 15 min and 24h after curing. RESULTS: Both the K(Ic) and HK increased with increased light energy density and storage time. Linear regression analysis indicated a strong correlation between HK and K(Ic) tested at the same time period (R(2)=0.97 and 0.90 for 15 min and 24h, respectively). The correlation became weaker between the different storage times (R(2)=0.71), indicating a change in fracture toughness and/or microhardness mechanisms. CONCLUSION: Fracture toughness of a composite cannot be simply extrapolated from microhardness. PMID- 12654548 TI - Volumetric shrinkage of composites using video-imaging. AB - OBJECTIVE: This study involves investigation of the use of video-imaging for measurement of volumetric shrinkage of composites. METHODS: Six composites were tested for volumetric shrinkage using video-imaging. The volumetric shrinkage was measured using the single- and multi-view volumetric reconstruction modes. All composites were cured using a VIP(TM) curing light for 40s at 500 mW/cm(2). Dynamic shrinkage was measured using the single-view mode with a red filter placed over the detector opening. RESULTS: Analysis of the volumetric shrinkage values by a one way ANOVA for each composite showed no difference for the single- and multi-view measurement mode. The shrinkage values determined by video-imaging were compared to those measured for the same composites by mercury dilatometry by one way ANOVA followed by a paired comparison using the Bonferroni method. CONCLUSION: The video-imaging technique gives reproducible results for volumetric shrinkage of composites comparable to those measured by dilatometry. PMID- 12654549 TI - Efficacy of anti-retraction devices in preventing bacterial contamination of dental unit water lines. AB - OBJECTIVE: In this study we examine the efficacy of anti-retraction devices in preventing microbial contamination of dental unit water lines (DUWLs). METHODS: The study was performed on 54 randomly selected DUs in use at private and public institutions for over six months. The selected DUs were all currently commercially available. To evaluate the efficiency of anti-retraction devices, two different methods were employed, mechanical and microbiological. The ADA/ANSI specification #47 (corresponding to a water retraction of less than 40.3 microl) was used for the evaluation of the retraction determined using the mechanical method; the presence/absence of test microorganisms in DUWLs upstream the turbine after the test was used for evaluating the microbiological assay. RESULTS: Both evaluation methods indicated correct amounts of retraction and prevention of DUWL contamination occurred in only two cases (3.7%). Correct retraction was noted in six other dental units (DUs), but their DUWLs became contaminated during testing. Six other produced the opposite results-improper retraction, yet no contamination. The remaining 40 DUs (74.0%) failed both evaluations. CONCLUSION: The results showed that the overwhelming majority of the anti-retraction devices did not prevent retraction when the turbine stopped running, leading to a contamination of the water lines, and to a consequent possible cross contamination of the patients. PMID- 12654550 TI - Comparison of antibacterial activity of two dentin bonding systems using agar well technique and tooth cavity model. AB - OBJECTIVE: This study compared the antibacterial activities of two dentin bonding systems (ABF, Kuraray and Reactmer Bond, Shofu) by a conventional agar well technique and a newly designed in vitro test using tooth model. METHODS: In the agar well technique, the test materials were filled in the wells of Muller Hinton agar plates inoculated with Streptococcus mutans NCTC10449, and the diameters of inhibition zones produced around the materials were measured after 24h of incubation. For the tooth model test, three cavities (diameter 1mm, depth 2mm) were prepared in the flat occlusal dentin of human extracted molar. After sterilization, the teeth were left in broth culture of 1.56 x 10(8)CFU/ml of S. mutans at 37 degrees C for 72h for allowing bacteria to invade the cavity. The dentin bonding systems were applied separately to each of the two infected cavities, and the third cavity was left unapplied for control. After sealing the occlusal surfaces, the teeth were kept in physiologic saline solution at 37 degrees C for 72h. The standardized amounts of dentin chips (120+/-5mg) were obtained from the cavity walls and the number of bacteria recovered was determined. The results were analyzed by One Way ANOVA, Kruskal Wallis and Mann Whitney's U tests. RESULTS: The primer of ABF and Reactmer Bond produced inhibition zones with similar sizes (p>0.05), but the bonding resin of ABF did not produce any inhibition. When tested by the model cavity method, the application of ABF resulted in significantly less bacterial recovery than Reactmer Bond (p<0.05), demonstrating substantial antibacterial effects. CONCLUSIONS: The tooth model method used in this study was effective for evaluating the substantial antibacterial effects of dentin bonding agents, and the experimental dentin bonding system ABF was demonstrated to be able to inactivate the bacteria in the cavity effectively in comparison with little antibacterial activity shown by Reactmer Bond. PMID- 12654551 TI - Bond failure at dentin-composite interfaces with 'single-bottle' adhesives. AB - OBJECTIVES: Significant differences in interfacial fracture toughness (intK(1c)) among six dentinal adhesives have been reported. Resulting fractured test specimens were examined under SEM to determine differences in micromorphology, which might account for the respective intK(1c) results. METHODS: Interfacial fracture toughness specimens were assembled from bovine dentin and P50 resin composite under moist conditions, using one of five 'single-bottle' adhesives; either Single Bond, One Step, Optibond Solo, Prime & Bond 2.1, Bond 1 or a resin modified glass ionomer, Fuji Bond LC. After fracture toughness testing, four fractured specimens from each group were sectioned transversely, critical-point dried and examined under SEM. RESULTS: Most bond failures occurred at the interface between adhesive resin and the top of the hybrid layer. Single Bond and One Step gave the highest intK(1c) results and showed good resin infiltration at this interface. Average film thickness of unfilled adhesives was 30 microm. When this intermediate adhesive layer was too thin, resin infiltration was poor and associated with low fracture toughness. Filled adhesives, Optibond Solo and Fuji Bond LC, formed thick films varying from 60 to 250 microm which failed cohesively, effectively sealing the dentin surface despite moderate fracture toughness results. CONCLUSIONS: Most bond failures occurred between the adhesive and hybrid layers. Good resin infiltration at the top of the hybrid layer combined with an intermediate adhesive layer of adequate width produced a fracture-resistant interface. Filled adhesives failed cohesively, providing a good dentin seal, despite material fracture. PMID- 12654552 TI - SEM evaluation of the interaction pattern between dentin and resin after cavity preparation using ER:YAG laser. AB - OBJECTIVE: The aim of this study was to describe the interaction pattern formed between dentin and resin on cavities prepared with an erbium laser (Er:YAG). The morphological aspect of the irradiated dentin after acid etching was also observed. METHODS: Ten dentin disks were obtained from fresh extracted third molars. Each disk received two cavities, one prepared with a conventional high speed drill, while the other cavity was obtained by the use of an Er:YAG laser (KaVo KEY Laser, KaVo Co.). The laser treatment was performed with 250 mJ/pulse, 4Hz, non contact mode, focused beam, and a fine water mist was used. Five disks were prepared for morphological analysis of the acid etched dentin. The other five disks had their cavities restored with Single Bond (3M) followed by Z100 resin (3M). The specimens were observed under scanning electron microscopy after dentin-resin interface demineralization and deproteinization. RESULTS AND CONCLUSIONS: It was observed that the morphological characteristics of the acid etched irradiated dentin were not favorable to the diffusion of monomers through the collagen network. The dentin-resin interfacial aspect of irradiated dentin, after acid etching, showed thin tags and scarce hybridization zones, which agreed with the morphology of the irradiated and acid-etched dentin substrate observed. PMID- 12654553 TI - Acidic solubility of luting cements. AB - OBJECTIVES: Chemical disintegration of luting cements can adversely affect their long term success. The aim of the present study was to assess the susceptibility of zinc phosphate cement, glass ionomer cement and resin cement to erosion at various pH values. METHODS: Zinc phosphate and glass ionomer cement samples were eroded in 0.3% citric acid adjusted to pH 3.0, 4.0, 5.0, 6.0 and 7.0 at a temperature of 35 degrees C. A control group was stored in saline. Profilometric readings were performed after 30 min and 1h erosion time. Resin cement samples were eroded at pH 3.0 with measurements after 30 min, 6h, 24h and 7 days. A saline control group was measured after 7 days. RESULTS: Erosion of zinc phosphate cement was significantly higher at all pH values than erosion of glass ionomer cement with erosion depth increasing with decreasing citric acid pH. No erosion was seen in the control groups. The resin cement did not show any change in surface profile after the 7 day erosion period compared to the control group. Comparison to dental tissues revealed that erosion of glass ionomer cement in vitro was slightly lower than erosion of enamel and dentine. In contrast, erosion of zinc phosphate cement was much higher. PMID- 12654554 TI - Restorative service trends in private general practice in Australia: 1983-1999. AB - OBJECTIVES: Restorative services are a major component of the annual workload of a dentist, but are undergoing a period of change as population oral health continues to improve and restorative materials and techniques are developed. The aim of this study was to examine trends in the provision of restorative services over a 15-year period spanning 1983-84 to 1998-99. METHODS: A series of cross sectional surveys was performed in 1983-84, 1988-89, 1993-94 and 1998-99 with response rates of over 70%. Data were collected by a mailed survey from a random sample of dentists from each state/territory in Australia. Dentists provided data on patients treated on a typical clinical day from which annual rates of provision of services per dentist were derived for restorative and related services. RESULTS: The overall number of annual restorative services provided per dentist declined over the four points of the study period. This trend reflected a steady decline in the annual number of amalgams provided per dentist. However, composite resins increased between 1993-94 and 1998-99, while glass ionomers, other-restorative services, crowns and fissure sealants all showed increases over the first three points of the study period. CONCLUSIONS: Over the study period the highest provision of restorative and related services changed from amalgams to composite resins, reflecting trends towards improved oral health and development of restorative materials. The decline in restorative services was not uniform across all age groups of patients, with increases observed among 45-64 and 65+ year olds, reflecting increased retention of teeth among adults. PMID- 12654555 TI - The efficacy of a toothbrush disinfectant spray--an in vitro study. AB - OBJECTIVES: Toothbrushes are contaminated by micro-organisms which may be sources of infection, re-infection or cross infection. The object of this research is to test the efficacy of a proprietary toothbrush spray disinfectant (Brushtox) against specific test bacteria and fungi and to consider its potential. METHODS: Three bacteria and two fungi representing a broad microbial spectrum with relevance to oral bacteria were used in four laboratory tests including an in vitro toothbrush disinfection test. RESULTS: Brushtox proved to be almost 100% effective in three tests and showed significant inhibitory action in the standard agar diffusion test. CONCLUSION: Brushtox is an effective disinfectant agent for bacteria and fungi on toothbrushes. The solution may be valuable in the control of a wide variety of oral infections and secondary infections, especially in frail or debilitated individuals or those on immuno-suppressive therapy, and also in healthy individuals. PMID- 12654556 TI - Volumetric analysis predicts hepatic dysfunction in patients undergoing major liver resection. AB - Liver-enhancing modalities, such as portal vein embolization, are increasingly employed prior to major liver resection to prevent postoperative liver dysfunction. Selection criteria for such techniques are not well described. This study uses CT-based volumetric analysis as a tool to identify patients at highest risk for postoperative hepatic dysfunction. Between July 1999 and December 2000, a total of 126 consecutive patients who were undergoing liver resection for colorectal metastasis and had CT scans at our institution were included in the analysis. Volume of resection was determined by semiautomated contouring of the liver on preoperative volumetrically (helical) acquired CT scans. Hepatic dysfunction was defined as prothrombin time greater than 18 seconds or serum bilirubin level greater than 3 mg/dl. Marginal regression was used to compare the predictive ability of volumetric analysis and the extent of resection. The percentage of liver remaining was closely correlated with increasing prothrombin time and bilirubin level (P < 0.001). After trisegmentectomy, 90% of patients with .2 mm), three had micrometastases (between 2 mm and 0.2 mm), and five had isolated tumor cells or clusters (<.2 mm) identified by immunohistochemical analysis only. Patients had significantly fewer blue/radioactive ("hot") nodes than blue-only nodes (1.38 vs. 2.48 per patient; P = 0.0001). It is important to note that nodal metastases were more common in blue/hot nodes than in blue-only nodes (27.3% [19 of 68] vs. 8.8% [11 of 124]; P = 0.005). Dual-agent lymphatic mapping more accurately identifies sentinel node metastases than blue dye alone. In addition, this technique allows a more focused histopathologic examination of these nodes, in conjunction with the revised American Joint Committee on Cancer guidelines, and thereby offers the potential for significant upstaging of CRC. PMID- 12654560 TI - Inhibition of the phosphatidylinositol 3'-kinase signaling pathway increases the responsiveness of pancreatic carcinoma cells to sulindac. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) may be effective treatment for pancreatic cancer. We have previously demonstrated that NSAIDs suppress pancreatic cell growth in vitro by inhibiting cell cycle progression but have little effect on apoptosis. In fact, we have shown that NSAIDs, in some instances, increase Akt phosphorylation in human pancreatic carcinoma cells suggesting activation of the phosphatidylinositol 3'-kinase (PI3K)-Akt survival (antiapoptotic) pathway. We subsequently examined the effects of treating the human pancreatic cancer cell lines BxPC-3 and PaCa-2 with a specific inhibitor of the PI3K/Akt pathway, LY294002, in the presence or absence of the NSAID sulindac. The growth effects of sulindac (250 to 500 micromol/L) and/or LY294002 (1 to 100 micromol/L) were determined by a colorimetric proliferation assay and cell counts. The combination of low-dose LY294002 (10 micromol/L) and sulindac enhanced the growth inhibitory effects of sulindac in BxPC-3 and PaCa-2 cells. Treatment of both cell lines with the LY294002/sulindac combination altered the cell cycle distribution as determined by flow cytometry and also lowered the apoptotic threshold as measured with an enzyme-linked immunosorbent assay to detect DNA fragmentation. These effects were associated with changes in the expression and/or phosphorylation level of proteins and kinases that regulate cell cycle progression and apoptosis. Taken together, our results suggest that inhibition of the PI3K/Akt signaling pathway may sensitize pancreatic tumor cells to therapy with NSAIDs such as sulindac. PMID- 12654559 TI - Small bowel extrinsic denervation does not alter water and electrolyte absorption from the colon in the fasting or early postprandial state. AB - Small bowel transplantation (SBT) causes watery diarrhea. The decreases shown previously in absorption of water, electrolytes, and bile salts in the jejunum and ileum, although present, are not dramatic and seem not to be great enough to explain the diarrhea. Our aim was to determine changes in water and electrolyte absorption in the colon during fasting and postprandially in a canine preparation of jejunoileal extrinsic denervation, which serves as a model of jejunoileal autotransplantation. We hypothesized that colonic absorption of water and electrolytes decreases transiently in the colon after SBT. Six dogs had cannulas implanted in the colon to study absorption of an ileal-like basal electrolyte solution with or without 10 mmol/L glucose. Absorption during fasting and postprandially was measured before and 2 and 12 weeks after a validated preparation of jejunoileal extrinsic denervation. All dogs developed diarrhea after SBT. Net colonic absorptive fluxes of water and electrolytes in the colon did not change from baseline values at 2 or 12 weeks after extrinsic denervation, either during fasting or postprandially; glucose in the infusate did not alter absorptive fluxes during fasting or postprandially. Extrinsic denervation of the small intestine does not appear to alter colonic absorption of water or electrolytes during fasting or postprandially. These observations suggest that the neurally intact colon has a minimal role in the diarrhea after SBT. PMID- 12654561 TI - Development of a therapeutic adenoviral vector for cholangiocarcinoma combining tumor-restricted gene expression and infectivity enhancement. AB - Cholangiocarcinoma is an invasive malignancy that is most often unresectable upon diagnosis and unresponsive to chemotherapy and radiation. While adenoviral gene therapy has shown promise in treating many tumors, systemic toxicity and low tumor transduction efficiency have hampered its application in many gastrointestinal cancers. To overcome these difficulties, we have constructed an adenoviral vector utilizing a tumor-specific promoter (TSP) for selective transgene expression and a vector with an RGD-motif in the fiber-knob region for infectivity enhancement. In seeking a TSP for cholangiocarcinoma, Secretory Leukoprotease Inhibitor, Midkine, Gastrin Releasing Peptide, VEGF, Cox-2M, and Cox-2L promoters were configures in adenoviral vectors, and evaluated in cholangiocarcinoma cells lines (Oz and SkChA-1). Luciferase assays demonstrated that Cox-2 promoters (M and L) showed the highest promoter activity, with Cox-2M appearing slightly stronger than Cox-2L. Infectivity enhanced vectors with RGD motif in the fiber-knob region were also constructed with the luciferase transgene driven by a CMV control and the Cox-2M and Cox-2L promoters. Subsequent luciferase assays comparing the unmodified vectors to the RGD-modified versions demonstrated higher levels of luciferase activity than the RGD-infected cells. This paradigm was then applied to a therapeutic HSV-TK/GCV model by constructing RGD-enhanced HSV-TK vectors driven by Cox-2M and Cox-2L promoters. In vitro cytocidal effect analysis confirmed that the RGD-modified, cox-2 (M and L) driven vectors showed a stronger cytocidal effect upon gancyclovir administration than the vectors with wild-type fiber. The Cox-2 promoter demonstrates a favorable selectivity profile for cholangiocarcinoma, and RGD-modification further enhances transduction efficiency. This combination has potential to overcome the obstacles to clinical application of adenoviral gene therapy in cholangiocarcinoma. PMID- 12654562 TI - Incidence and management of biliary pancreatitis in cholecystectomized patients. Results of a 7-year study. AB - Data are lacking concerning the frequency of biliary acute pancreatitis in the postcholecystectomy patient. The aim of this study was to identify patients at risk for biliary pancreatitis after cholecystectomy and to describe the therapeutic management of these patients, based on an analysis of 278 unselected patients with acute pancreatitis during a 7-year period. A biliary etiology was presumed in the presence of laboratory findings of cholestasis that could not be explained by another disease, together with the absence of any other known etiology of acute pancreatitis. A biliary cause of disease was found in 132 (47%) of 278 patients. Seventeen (13%) of 132 patients had a history of cholecystectomy. Endoscopic retrograde cholangiopancreatography was performed in all patients with a suspected biliary cause of acute pancreatitis. It showed bile duct stones, microlithiasis, or sludge in 14 patients, and was consistent with typical findings at the papilla of Vater after stone passage in another three patients. No surgical bile duct exploration was necessary. One patient with severe disease and infected pancreatic necrosis died of septic multiorgan failure. PMID- 12654563 TI - Arginase acts as an alternative pathway of L-arginine metabolism in experimental colon anastomosis. AB - L-Arginine is the substrate for the nitric oxide synthase (NOS) pathway that is essential for gastrointestinal wound healing. L-Arginine is also the substrate for the enzyme arginase which metabolizes L-arginine to ornithine and subsequently to proline and polyamines both known to interact in cell proliferation and collagen synthesis. Two distinct isoforms of arginase exist. The temporal expression of the L-arginine metabolism in experimental colon anastomosis was investigated. Male Lewis rats underwent laparotomy. A left-sided colotomy was performed and the colon reanastomosed using 6-0 prolene. Sham operation was performed in controls. On days 2, 5, 10, 14, and 28 after the surgery the anastomosis was excised. The tissue at the anastomosis (ANAST) as well as above and below the anastomosis (PDC) and from sham colon was harvested and analyzed for distinct arginase isoform I (AI) and arginase isoform II (AII) activity, protein and mRNA expression as well as immunohistochemistry. iNOS protein and mRNA expression were investigated in parallel. A mean of 3 to 4 separate rats were analyzed per time point. Statistical analysis was performed by student's t-test, significance was reached when P < 0.05. AI activity, protein, and mRNA expression were significantly upregulated at the anastomosis compared to sham controls and PDC colons at all time points. The maximum was achieved at days 10 to 14 after wounding, and decreased to baseline levels thereafter. Inflammatory cells stained positive for AI. AII protein was not detectable. However RT-PCR showed low baseline expression. iNOS expression was upregulated early but for a shorter time period after wounding and reverted quickly to undetectable levels. In anastomotic healing, AI upregulation suggests a prolonged metabolism of arginine via arginase to polyamines and proline to provide substrate for collagen synthesis and cell proliferation. The functional implication of this arginase pathway further needs to be elucidated. PMID- 12654564 TI - Combined endoscopic/laparoscopic intragastric resection of gastric stromal tumors. AB - Myogenic neoplasms of the stomach are the most common submucosal mass. Their natural history is indeterminate, and surgical resection is advised regardless of size. These lesions have typically required open resection, but a variety of laparoscopic techniques have been described. We report results of endoscopically guided, laparoscopic intragastric resection. Fourteen lesions have been excised in 13 patients in the last 3.5 years. There were eight women and five men with a mean age of 57 years (range 34-72). All patients were asymptomatic, and no lesions had mucosal ulceration. Eight lesions were located at the gastroesophageal junction, two each at the incisura and posterior body, and one each in the fundus and anterior wall of the corpus. All lesions were predominantly intraluminal, and three were transmural. The diagnosis of a myogenic lesion was confirmed by endoscopic ultrasound in eight patients. The laparoscopic/endoscopic technique included two or three, 2 or 5 mm intragastric trocars; endoscopic suture passage and specimen removal; and laparoscopic intragastric suture repair of the gastric defect. The mean operative time was 186 minutes. The mean size of the resected specimens was 3.8 cm (range 1.5-7.0). There was no mitotic activity on histopathology, and all were considered pathologically benign. The median length of stay was 3.8 days (range 3-8). There was no mortality or operative morbidity. At a mean follow-up of 16.2 months (range 1-32) there has been no local recurrences. A combined laparoscopic/endoscopic intragastric resection is most appropriate for intraluminal, benign-appearing submucosal lesions of the proximal stomach. PMID- 12654565 TI - Modulation of growth factor and cytokine expression by nitric oxide during rat colon anastomotic healing. AB - We have previously shown that inhibition of nitric oxide generated by inducible nitric oxide synthase (iNOS) results in impaired colon anastomotic healing. Therefore, we proceeded to assess whether disruption of iNOS activity alters the normal pattern of growth factor expression during anastomotic healing. Two groups of male Sprague-Dawley rats underwent distal colonic division and anastomosis, jugular venous catheterization and subcutaneous placement of polyvinyl alcohol sponges. The first group (n = 10) received q8 hour intravenous injections of 10 mg/kg L-N-iminoethyl-lysine (L-NIL, a selective inhibitor of iNOS), while the second group (n = 12) received equal volumes of saline. On postoperative day 5, animals were sacrificed and anastomotic bursting pressure was determined. Histologic sections of the anastomosis were subjected to in situ hybridization versus mRNA of the proteins listed below. Positive controls were reacted with a poly-thymidine (poly-T) probe versus ubiquitous mRNA poly-adenine tails. Positively stained cells were quantified using a calibrated optical grid encompassing 0.5 mm(2) area centered over the anastomosis. Results are reported as the number of positive cells per 1000 cells positive for poly-T. L-NIL treated animals demonstrated an 18% decrease in wound fluid NO(X) compared to controls (29.2 +/- 1.2 vs. 34.6 +/- 2.0 microM, mean +/- SEM; P = 0.035). This corresponded to a 17% decrease in anastomotic bursting pressure (153 +/- 4 vs. 182 +/- 8 mm Hg, mean +/- SEM; P < 0.05). L-NIL also markedly increased the number of cells expressing transforming growth factor-beta, tumor necrosis factor alpha, vascular endothelial growth factor, and both inducible and endothelial forms of nitric oxide synthase. L-NIL had no effect on the expression of basic fibroblast growth factor. The data demonstrate that iNOS inhibition markedly disrupts the profile of cytokine and growth factor mRNA normally expressed during anastomotic healing. This provides in vivo evidence that NO modulates gene expression during anastomotic healing. PMID- 12654568 TI - Bile duct injuries 12 years after the introduction of laparoscopic cholecystectomy. PMID- 12654567 TI - Treatment strategies for hepatocellular carcinoma in cirrhosis. PMID- 12654569 TI - Primary cystic neoplasms of the pancreas. Neoplastic disorders of emerging importance-current state-of-the-art and unanswered questions. PMID- 12654570 TI - Roux-en-Y gastric bypass after previous unsuccessful gastric restrictive surgery. PMID- 12654572 TI - Understanding congestive heart failure (CHF). PMID- 12654573 TI - Music as medicine. PMID- 12654574 TI - All in the family: heredity and prostate cancer. PMID- 12654576 TI - Are antidepressants placebos? A persistent controversy could affect the choice of psychiatric symptoms. PMID- 12654575 TI - I am an avid long-distance biker, but I've noticed some tingling in my penis after long rides. What can I do to prevent problems with my sex life? PMID- 12654577 TI - Anorexia nervosa. Part II. PMID- 12654578 TI - Varieties of schizophrenia. PMID- 12654579 TI - A genetic defense against child abuse? PMID- 12654580 TI - Hats off to an old blood pressure drug. For treating high blood pressure, an old- and inexpensive--medication is a great place to start or a good one to add. PMID- 12654581 TI - Should you see about your CRP? A simple blood test for C-reactive protein is joining cholesterol screening as an early warning for heart disease. But not everyone needs it. PMID- 12654582 TI - No age limit for cholesterol-lowering benefits. Older people gain as much as younger people from cholesterol-lowering statins. PMID- 12654584 TI - Don't let drug names confuse you. PMID- 12654583 TI - Slow? Or steady? Both win the race. For taming atrial fibrillation, controlling the heart's rate or its rhythm work equally well. Either way, it's important to keep taking drugs that prevent blood clots. PMID- 12654585 TI - Smart pacemakers join the Web. PMID- 12654586 TI - Anti-inflammatory drugs may put on the pressure. PMID- 12654587 TI - Ask the doctor. I took the diet drug sibutramine (Meridia) for two months but stopped when I heard that its use had been linked to some deaths. During this time I developed mitral valve regurgitation. Could this have been a side effect of Meridia? PMID- 12654588 TI - Ask the doctor. At age 62 I'm a bit overweight and have diabetes. I take a statin, and my LDL cholesterol is good (84 mg/dL). But my HDL is low (30 mg/dL) and my triglycerides are above 300 mg/dL. Are high triglycerides a problem? PMID- 12654589 TI - Which blood pressure drug is best for you? If you need medication to control your blood pressure, it may be reassuring to know that the best choice is also the cheapest. PMID- 12654590 TI - Forestalling frailty. The average woman can now expect live into her 80s. Whether she enjoys her later years may depend on avoiding a common, though not inevitable, consequence of aging. PMID- 12654591 TI - Healthy mouth, healthy life. New understanding of the relationship between oral health and general health bolsters the tried-and-true advice to brush, floss, and see your dentist often. PMID- 12654592 TI - Dry mouth: more than just an annoyance. PMID- 12654593 TI - Costochondritis: Not a heart attack but it feels like one. PMID- 12654594 TI - Readers question waist whereabouts. PMID- 12654595 TI - By the way, doctor. I recently read that the new osteoporosis drug Forteo is better than any available alternative. Since so many of us who are postmenopausal are at risk for osteoporosis, shouldn't we be taking it? PMID- 12654596 TI - By the way, doctor. The more I eat a health diet--one that's rich in whole grains, fruits, and vegetables--the more trouble I have with flatulence. Is this unusual? Any suggestions? PMID- 12654597 TI - A guide for the perplexed: towards an understanding of the molecular basis of heart failure. PMID- 12654599 TI - Regionalization of care for acute ischemic heart disease: a call for specialized centers. PMID- 12654598 TI - Proinflammatory cytokines: predictors of a failing heart? PMID- 12654600 TI - Need for centers to care for patients with acute coronary syndromes. PMID- 12654601 TI - Editor's commentary: centers of excellence. PMID- 12654602 TI - Family history and cardiovascular risk in familial hypercholesterolemia: data in more than 1000 children. AB - BACKGROUND: Elevated LDL cholesterol (LDL-C) levels in childhood predict cardiovascular disease (CVD) later in life. Familial hypercholesterolemia (FH) represents the paradigm of this relation. METHODS AND RESULTS: The objectives of this study were to (1) establish the LDL-C level that provides the most accurate diagnosis of FH in children from families with known FH and (2) assess whether lipoprotein variation in these children is associated with premature CVD in relatives. Foremost, however, it was our objective to identify children with FH who are at high risk and in need of early intervention. A total of 1034 consecutive children from FH kindreds were investigated. First, LDL-C levels >3.50 mmol/L had a 0.98 post-test probability (95% CI, 0.96 to 0.99) of predicting the presence of an LDL receptor mutation. Second, children with FH in the highest LDL-C tertile (>6.23 mmol/L) had a 1.7-times higher incidence (95% CI, 1.24 to 2.36) of having a parent with FH suffering from premature CVD (P=0.001). In addition, such a parent was found 1.8 times more often (95% CI, 1.20 to 2.59) among children with FH who had HDL-C <1.00 mmol/L (P=0.004). Last, children with FH whose lipoprotein(a) was >300 mg/L had a 1.45-times higher incidence (95% CI, 0.99 to 2.13) of having a parent with FH suffering from premature CVD (P=0.05). CONCLUSIONS: In FH families, LDL-C levels allow accurate diagnosis of FH in childhood. Moreover, increased LDL-C and lipoprotein(a) and decreased HDL-C levels in children identify FH kindreds with the highest CVD risk. PMID- 12654603 TI - Suppression of the functionally coupled cyclooxygenase-2/prostaglandin E synthase as a basis of simvastatin-dependent plaque stabilization in humans. AB - BACKGROUND: The clinical benefits of statins are attributed to changes in plaque composition that lead to reduced metalloproteinase (MMP) activity and plaque stabilization. However, the molecular mechanism of this effect is unclear. Recently, we demonstrated enhanced expression of isoforms of inducible cyclooxygenase (COX) and PGE synthase (COX-2/mPGES) in human symptomatic plaque and provided evidence that this is associated with MMP-induced plaque rupture. The aim of this study was to characterize the effect of simvastatin on inflammatory infiltration and the expression of COX-2/mPGES and MMPs in human carotid plaques. METHODS AND RESULTS: Seventy patients with symptomatic carotid artery stenosis were randomized to the American Heart Association Step 1 diet plus simvastatin (40 mg/d) or the American Heart Association Step 1 diet alone for 4 months before endarterectomy. Plaques were subjected to analysis of COX-1, COX-2, mPGES, MMP-2 and MMP-9, lipid and oxidized LDL (oxLDL) content, and collagen content by immunocytochemistry, Western blot, and reverse transcription polymerase chain reaction, whereas zymography was used to detect MMP activity. Immunocytochemistry was also used to identify CD68+ macrophages, CD3+ T lymphocytes, smooth muscle cells (SMCs), and HLA-DR+ inflammatory cells. Plaques from the simvastatin group had fewer (P<0.0001) macrophages, T-lymphocytes, and HLA-DR+ cells; less (P<0.0001) immunoreactivity for COX-2/mPGES and MMPs; reduced (P<0.0001) gelatinolytic activity; increased (P<0.0001) collagen content; and reduced (P<0.0001) lipid and oxLDL content. Interestingly, COX-2/mPGES inhibition by simvastatin was completely reversed by mevalonate in vitro. CONCLUSIONS: This study demonstrates that simvastatin decreases inflammation and inhibits COX 2/mPGES expression in plaque macrophages, and this effect in turn may contribute to plaque stabilization by inhibition of MMP-induced plaque rupture. PMID- 12654604 TI - Inflammatory markers and risk of heart failure in elderly subjects without prior myocardial infarction: the Framingham Heart Study. AB - BACKGROUND: Experimental studies support a key role for cytokines in left ventricular remodeling. In congestive heart failure (CHF) patients, elevated plasma cytokine levels are associated with worse functional status and adverse prognosis. It is unclear whether cytokine levels can predict the incidence of CHF in asymptomatic individuals. METHODS AND RESULTS: We investigated the relations of serum interleukin-6 (IL-6), C-reactive protein (CRP), and spontaneous production of tumor necrosis factor-alpha (TNFalpha) by peripheral blood mononuclear cell (PBMC) to CHF incidence among 732 elderly Framingham Study subjects (mean age 78 years, 67% women) free of prior myocardial infarction and CHF. On follow-up (mean 5.2 years), 56 subjects (35 women) developed CHF. After adjustment for established risk factors, including the occurrence of myocardial infarction on follow-up, there was a 60 (PBMC TNFalpha) to 68% (serum IL-6) increase in risk of CHF per tertile increment in cytokine concentration (P=0.04, and 0.03, respectively, for trend). A serum CRP level > or =5 mg/dL was associated with a 2.8-fold increased risk of CHF (P=0.02). Subjects with elevated levels of all 3 biomarkers (serum IL-6 and PBMC TNFalpha >median values, CRP> or =5 mg/dL) had a markedly increased risk of CHF (hazards ratio 4.07 [95% CI 1.34 to 12.37], P=0.01) compared with the other subjects. CONCLUSIONS: In our elderly, community-based sample, a single determination of serum inflammatory markers, particularly elevated IL-6, was associated with increased risk of CHF in people without prior myocardial infarction. Additional epidemiological investigations are warranted to confirm the contribution of inflammation to the pathogenesis of CHF in the general population. PMID- 12654605 TI - Mitral annular calcification predicts cardiovascular morbidity and mortality: the Framingham Heart Study. AB - BACKGROUND: Mitral annular calcification (MAC) has been associated with stroke in longitudinal, community-based cohorts and cardiovascular disease (CVD) outcomes in many small retrospective studies. Prospective data are limited on the relation of MAC with CVD morbidity and mortality. METHODS AND RESULTS: We examined the association between MAC assessed by M-mode echocardiography and the incidence of CVD, CVD death, and all-cause death over 16 years of follow-up in the Framingham Heart Study subjects who attended a routine examination between 1979 and 1981. Cox proportional hazards models were used to estimate hazard ratios (HRs) associated with the presence of MAC for each outcome. Of 1197(445 male, 752 female) subjects who had adequate echocardiographic assessment, 14% had MAC. There were 307 incident CVD events and 621 deaths. In multivariable adjusted analyses, MAC was associated with an increased risk of incident CVD (HR, 1.5; 95% CI, 1.1, 2.0), CVD death (HR, 1.6; 95% CI, 1.1, 2.3), and all-cause death (HR, 1.3; 95% CI, 1.04, 1.6). For each 1-mm increase in MAC, the risk of incident CVD, CVD death, and all-cause death increased by approximately 10%. CONCLUSIONS: The independent association of MAC with incident CVD and CVD death underscores that cardiac calcification is a marker of increased CVD risk. PMID- 12654606 TI - Adjunctive platelet glycoprotein IIb/IIIa receptor inhibition with tirofiban before primary angioplasty improves angiographic outcomes: results of the TIrofiban Given in the Emergency Room before Primary Angioplasty (TIGER-PA) pilot trial. AB - BACKGROUND: Previous work has suggested that platelet glycoprotein IIb/IIIa receptor blockade may confer benefit in the treatment of acute myocardial infarction. The TIGER-PA pilot trial was a single-center randomized study to evaluate the safety, feasibility, and utility of early tirofiban administration before planned primary angioplasty in patients presenting with acute myocardial infarction. METHODS AND RESULTS: A total of 100 patients presenting with acute myocardial infarction were randomized to either early administration of tirofiban in the emergency room or later administration in the catheterization laboratory. The primary outcome measures were initial TIMI grade flow, corrected TIMI frame counts, and TIMI grade myocardial perfusion ("blush"). Thirty-day major adverse cardiac events were also assessed. Angiographic outcomes demonstrate a significant improvement in initial TIMI grade flow, corrected TIMI frame counts, and TIMI grade myocardial perfusion when patients are given tirofiban in the emergency room before primary angioplasty. The rate of 30-day major adverse cardiac events suggests that early administration may be beneficial. CONCLUSIONS: This pilot study suggests that early administration of tirofiban improves angiographic outcomes and is safe and feasible in patients undergoing primary angioplasty for acute myocardial infarction. PMID- 12654607 TI - Value of magnetic resonance imaging for the noninvasive detection of stenosis in coronary artery bypass grafts and recipient coronary arteries. AB - BACKGROUND: Magnetic resonance imaging (MRI) is a potential noninvasive diagnostic tool to detect coronary artery bypass graft stenosis, but its value in clinical practice remains to be established. We investigated the value of MRI in detecting stenotic grafts, including recipient vessels. METHODS AND RESULTS: We screened for inclusion 173 consecutive patients who were scheduled for coronary angiography because of recurrent chest pain after coronary artery bypass grafting (CABG). We studied 69 eligible patients with 166 grafts (81 single vein, 44 sequential vein, and 41 arterial grafts). MRI with baseline and stress flow mapping was performed. Both scans were successful in 80% of grafts. Grafts were divided into groups with stenosis > or =50% (n=72) and > or =70% (n=48) in the graft or recipient vessels. Marginal logistic regression was used to predict the probability for the presence of stenosis per graft type using multiple MRI variables. Receiver operator characteristics (ROC) analysis was performed to assess the diagnostic value of MRI. Sensitivity (95% confidence interval)/specificity (95% confidence interval) in detecting single vein grafts with stenosis > or =50% and > or =70% were 94% (86 to 100)/63% (48 to 79) and 96% (87 to 100)/92% (84 to 100), respectively. CONCLUSIONS: MRI with flow mapping is useful for identifying grafts and recipient vessels with flow-limiting stenosis. Flow scans could be obtained in 80% of the grafts. This proof-of-concept study suggests that noninvasive MRI detection of stenotic grafts in patients who present with recurrent chest pain after CABG may be useful in selecting those in need of an invasive procedure. PMID- 12654608 TI - Influence of inhaled iloprost on transpulmonary gradient of big endothelin in patients with pulmonary hypertension. AB - BACKGROUND: The pulmonary circulation is an important site for the production and clearance of endothelin (ET)-1, a potent vasoactive and mitogenic peptide. In healthy individuals, 40% to 50% of circulating ET-1 is removed on each passage through the lungs resulting in an arteriovenous ratio of <1, whereas many patients with pulmonary arterial hypertension (PAH) have ratios >1, indicating reduced clearance or increased release of endothelin. The influence of inhaled prostanoids on endothelin clearance is unknown. METHODS AND RESULTS: In a prospective investigation, plasma concentrations of big endothelin-1 (big ET-1, Elisa) were measured in 15 patients with pulmonary hypertension undergoing right heart catheterization with iloprost inhalation (4 m, 11 f, aged 35 to 75 years, mean pulmonary arterial pressure (PAPm) 54+/-2.3 mm Hg, pulmonary vascular resistance (PVR) 1061+/-141 dyn x sec x cm(-5)). There was a significant transpulmonary gradient for big ET-1 with 31% +/-11% higher concentrations in the radial artery than in the pulmonary artery (P<0.001). After inhalation of iloprost a significant decrease in the AV-ratio from 1.31+/-0.11 to 0.92+/-0.06 (P<0.007) was observed. The pulmonary net release of 3.10+/-0.65 pmol/min big ET 1 at baseline decreased to -1.24+/-1.32 pmol/min (P=0.013) within 15 minutes indicating a restored balance. Patients under long-term treatment with iloprost (n=7) tended to have a lower net release and AV-ratio for big ET-1 than patients without pretreatment. CONCLUSION: An increase in pulmonary clearance of big-ET could be a mechanism contributing to the beneficial effects of inhaled prostanoids in the treatment of PAH. PMID- 12654609 TI - Why are patients with chronic obstructive pulmonary disease at increased risk of cardiovascular diseases? The potential role of systemic inflammation in chronic obstructive pulmonary disease. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) increases the risk of cardiovascular disease 2- to 3-fold. The factors responsible for this association remain largely unknown. METHODS AND RESULTS: We analyzed data from participants, > or =50 years of age, of the Third National Health and Nutrition Examination Survey (n=6629) to determine whether C-reactive protein (CRP) and other systemic inflammatory markers are present in participants with chronic airflow obstruction and are associated with cardiac injury. Participants with severe airflow obstruction had circulating leukocyte, platelet, and fibrinogen levels that were 460/ microL (95% confidence interval [CI], 30 to 890/ microL), 39 510/ microL (95% CI, 21 730 to 57 290/ microL), and 41.63 mg/dL (95% CI, 19.87 to 63.39 mg/dL) higher, respectively, than in those without airflow obstruction. They were also 2.18 times (95% CI, 1.46 to 3.27) more likely to have an elevated circulating CRP level. Moderate airflow obstruction was associated with smaller but still significant increases in these levels. Moderate and severe airflow obstruction was associated with increased occurrence of ischemic changes on electrocardiograms of participants. In the presence of both highly elevated CRP and moderate or severe airflow obstruction, the Cardiac Infarction Injury Score was 2.68 and 5.88 U higher, respectively, than in those without airflow obstruction and with low CRP, which suggests an additive effect of CRP and COPD on the risk of cardiac injury. CONCLUSION: Low-grade systemic inflammation was present in participants with moderate to severe airflow obstruction and was associated with increased risk of cardiac injury. This may in part explain the high rates of cardiovascular complications in COPD. PMID- 12654610 TI - L-4F, an apolipoprotein A-1 mimetic, restores nitric oxide and superoxide anion balance in low-density lipoprotein-treated endothelial cells. AB - BACKGROUND: Low-density lipoprotein (LDL) impairs endothelial cell function by uncoupling endothelial nitric oxide synthase (eNOS) activity, which allows superoxide anion (O2*-)) to be generated rather than nitric oxide (*NO). Recent reports indicate that apolipoprotein (apo) A-1 mimetics inhibit the development of atherosclerotic lesions in LDL receptor-null mice. Here we hypothesize that L 4F, an apoA-1 mimetic that inhibits atherosclerosis induced by hypercholesterolemia, protects endothelial cell function by preventing LDL from uncoupling eNOS activity. METHODS AND RESULTS: Bovine aortic endothelial cells were incubated with LDL+/-L-4F, and changes in A23187-stimulated.NO and O2*- generation were determined by ozone chemiluminescence and superoxide dismutase inhibitable ferricytochrome c reduction, respectively. Western analysis of eNOS immunoprecipitates was used to determine effects of LDL and L-4F on heat shock protein 90 (hsp90) interactions with eNOS. LDL decreased.NO production and increased eNOS-dependent O2*- generation. Pretreatment of LDL with L-4F increased.NO and decreased O2*- generation. By itself, L-4F had no effect on O2*- but did increase *NO generation. Stimulation of endothelial cells incubated with LDL decreased the association of hsp90 with eNOS. Pretreatment of LDL with L-4F prevented a decrease in hsp90 association with eNOS and often enhanced association on stimulation. CONCLUSIONS: These data demonstrate that L-4F protects endothelial cell function by preventing LDL from uncoupling eNOS activity. L-4F allows endothelial cell to maintain coupled eNOS activity to generate.NO even in the face of atherogenic concentrations of LDL. PMID- 12654611 TI - Display of Fas ligand protein on cardiac vasculature as a novel means of regulating allograft rejection. AB - BACKGROUND: Fas ligand (FasL) is a potent death-inducing molecule with important functions in immune homeostasis and tolerance to self-antigens. The complex biological activities of FasL and its inefficient expression using conventional gene transfer approaches limit its use for immunomodulation to prevent allograft rejection. We have recently generated a chimeric FasL with core streptavidin (SA FasL) with potent apoptotic activity and designed a novel approach to display it on the surface of several cell types via biotinylation. We herein tested whether SA-FasL can also be displayed on vascular endothelial cells in the heart and examined its effect on graft survival after transplantation into syngeneic and allogeneic hosts. METHODS AND RESULTS: SA-FasL was efficiently displayed on the vasculature of BALB/c hearts with a half-life of 9 days in vivo. Transplantation of hearts displaying SA-FasL into syngeneic hosts resulted in indefinite graft survival without detectable toxicity to the grafts and hosts. In contrast, transplantation of allogeneic C57BL/10 hearts displaying SA-FasL into BALB/c recipients resulted in graft rejection, but in a delayed fashion as compared with control hearts (mean survival time=17.4+/-5 versus 9.6+/-1 days). Allograft survival was further extended to 21+/-2.6 and 24+/-3 days (P<0.05) by intravenous treatment of graft recipients with 1 dose of SA-FasL-decorated donor splenocytes on days 2 and 6 after transplantation, respectively. CONCLUSIONS: This study shows for the first time that exogenous proteins can be displayed on the endothelium of solid organs for therapeutic purposes. This approach provides a convenient and rapid means of displaying exogenous proteins on the surface of cells, tissues, and solid organs, with broad research and therapeutic implications. PMID- 12654612 TI - Small GTP-binding protein Rac is an essential mediator of vascular endothelial growth factor-induced endothelial fenestrations and vascular permeability. AB - BACKGROUND: Vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) induces both angiogenesis and vascular permeability. Although its angiogenic activity has been well characterized, the signaling pathways of VEGF induced permeability remain poorly understood. METHODS AND RESULTS: Using the mouse corneal micropocket assay, Miles assay, and a combination of cytochemical, electron microscopic, and biochemical assays, we demonstrate that VEGF-induced vascular leakage partly can be separated from its angiogenic activity. VEGF but not FGF-2 induced capillaries with a highly fenestrated endothelium, a feature linked with increased vascular permeability. A cell-permeable Rac antagonist (TAT RacN17) converted VEGF-induced, leaky vascular plexuses into well-defined vascular networks. In addition, this Rac mutant blocked formation of VEGF-induced endothelial fenestrations and vascular permeability but only partially inhibited angiogenesis. Studies on endothelial cell cultures further revealed that VEGF stimulated phosphorylation of VEGF receptor-2 (VEGFR-2), leading to activation of Rac as well as increased phosphorylation of phospholipase Cgamma (PLCgamma), protein kinase B (Akt), endothelial nitric oxide synthase (eNOS), and extracellular regulated kinase (Erk1/2). We further found that phosphatidylinositol-3-OH kinase (PI3K) acted upstream of Rac and Akt-eNOS in VEGF/VEGFR-2 signaling. CONCLUSIONS: Our findings indicate that the small GTP binding protein Rac is a key component in mediation of VEGF-induced vascular permeability but less so in neovascularization. This may have conceptual implications for applying Rac antagonists in treatment and prevention of VEGF induced vascular leakage and edema in connection with ischemic disorders. PMID- 12654613 TI - Insulin-stimulated cyclic guanosine monophosphate inhibits vascular smooth muscle cell migration by inhibiting Ca/calmodulin-dependent protein kinase II. AB - BACKGROUND: Insulin resistance is associated with vascular disease. Physiological concentrations of insulin inhibit cultured vascular smooth muscle cell (VSMC) migration in the presence of nitric oxide, and the failure to do so in insulin resistant states may aggravate vascular disease. We sought to determine the molecular mechanisms by which insulin inhibits VSMC migration. METHODS AND RESULTS: Insulin at 1 nmol/L stimulated cGMP production in cultured rat VSMCs that were induced to express inducible nitric oxide synthase (iNOS). VSMC migration was measured in a wound-closure assay, and the platelet-derived growth factor-AB (PDGF-AB)-stimulated component of VSMC migration after wounding was inhibited by insulin, 8-Br-cGMP, and 1-[N-0-bis(5-isoquinolinesulfonyl)-N-methyl L-tyrosyl]-4-phenylpiperazine (KN-62), a selective inhibitor of calcium/calmodulin-dependent protein kinase II (CaM kinase II). Wounding alone or incubating cells with only PDGF-AB stimulated CaM kinase II activity in an insulin- and 8-Br-cGMP-inhibitable manner. Transfecting VSMCs with a constitutively active CaM kinase II mutant blocked the inhibition by insulin of both wound-induced and wound plus PDGF-AB-induced VSMC migration. High intracellular Ca2+ ([Ca]i)-stimulated CaM kinase II activity was inhibited by 8 Br-cGMP by an okadaic acid-sensitive mechanism. CONCLUSIONS: We conclude that in cultured rat VSMCs expressing iNOS, insulin, via stimulation of cGMP production, inhibits both wound alone-induced and the PDGF-AB-stimulated component of VSMC migration by inhibiting CaM kinase II activity. cGMP inhibits CaM kinase II at a post-[Ca]i step by a protein phosphatase-dependent mechanism. PMID- 12654614 TI - Superparamagnetic iron oxide-based method for quantifying recruitment of monocytes to mouse atherosclerotic lesions in vivo: enhancement by tissue necrosis factor-alpha, interleukin-1beta, and interferon-gamma. AB - BACKGROUND: It has been found recently that the MRI contrast agent superparamagnetic iron oxide (SPIO) localizes to aortic atherosclerotic plaques. We therefore asked whether SPIO might be used to monitor monocyte recruitment into aortic atherosclerotic plaques. METHODS AND RESULTS: Eleven female apo E knockout (K/O) mice, each 11 months old, were divided into 2 groups. Six mice received tissue necrosis factor-alpha (0.2 microg IP once), interleukin-1beta (0.2 micro g IP once), and interferon-gamma (100 U/g per day IP for 5 days); 5 received 0.5 mL saline containing 1% BSA and served as sham-treated atherosclerotic controls. Two wild-type C57BL/6 mice served as sham-treated nonatherosclerotic controls. Three hours after initial cytokine or sham treatment, all mice received SPIO by intravenous injection (1 mmol/kg iron). Six days later, all mice were euthanized, the hearts and aortas were perfused under physiological pressure, and the entire aortas were studied histologically. Atherosclerotic plaques in cytokine-treated mice contained more iron-positive macrophages per cross section than did those in sham-treated apo E K/O control mice (42+/-11.8 versus 11.6+/-5.9) (P<0.0001). Iron-laden macrophages were present either in subendothelial plaque surfaces or in thin layers overlying the internal elastic lamina, often at the edges of atherosclerotic plaques. No iron deposition was seen in aortas of the wild-type nonatherosclerotic control mice. Immunocytochemistry showed mostly macrophages and few T lymphocytes in atherosclerotic plaques of cytokine-treated mice. CONCLUSIONS: SPIO allows detection of iron-laden macrophages in the aortic subendothelium of apo E deficient mice under basal conditions and monitoring of monocyte recruitment after cytokine injection. PMID- 12654615 TI - Correlation between anatomy and electrical activation in canine pulmonary veins. AB - BACKGROUND: The roles of complex muscle sleeve geometry and fiber orientation in the pulmonary veins (PVs) in wave-front propagation are poorly understood. METHODS AND RESULTS: We mapped the left superior PV (LSPV, n=7) and left inferior PV (LIPV, n=4) of dogs with 420 bipolar electrodes (1-mm resolution) and performed detailed histological examination. In the anterior LSPV-left atrial (LA) junction, myocardial muscle fibers were oriented perpendicular to PV blood flow. A wedge filled with connective tissues led to a complete muscle separation or an abrupt increase in muscle thickness between the PV and LA (0.42+/-0.12 versus 2.0+/-0.31 mm, P<0.01). Distal LSPV pacing resulted in conduction block at the anterior PV-LA junction, with double potentials. In contrast, the posterior LSPV-LA junction showed gradual muscle thickening and a fiber orientation parallel to the blood flow. The maximum PV muscle thickness in the anterior PV-LA junction is thinner than that in the posterior junction (0.83+/-0.15 versus 1.3+/ 0.38 mm, P<0.01). Distal LIPV pacing showed multiple PV-LA breakthroughs, with segmental conduction block in the anterior PV-LA junction. The conduction block corresponded to segmental PV-LA muscle disconnection. Complex fiber orientations in the PV muscle sleeves away from the PV-LA junction were responsible for intra PV conduction delay or block during rapid PV pacing. CONCLUSIONS: We conclude that segmental muscle disconnection and differential muscle narrowing at PV-LA junctions and complex fiber orientations within the PV provide robust anatomical bases for conduction disturbance at the PV-LA junction and complex intra-PV conduction patterns. PMID- 12654616 TI - Important triad in cardiovascular medicine: diabetes, coronary intervention, and platelet glycoprotein IIb/IIIa receptor blockade. PMID- 12654617 TI - Images in cardiovascular medicine. Echocardiographic diagnosis of anomalous drainage of the superior vena cava into the left atrium. PMID- 12654618 TI - American Heart Association guidelines for primary prevention of atherosclerotic cardiovascular disease beginning in childhood. PMID- 12654619 TI - Images in cardiovascular medicine. Non-life-threatening leaflet escape. PMID- 12654620 TI - Titin stiffness in heart disease. PMID- 12654621 TI - Shouldering the risk burden: infection, atherosclerosis, and the vascular endothelium. PMID- 12654622 TI - Pretreatment of patients with atrial fibrillation who undergo electrical cardioversion. PMID- 12654623 TI - Low serum levels of soluble RANK ligand are associated with the presence of coronary artery disease in men. PMID- 12654624 TI - Why does elevated plasma homocysteine result in severe microvascular injury, but not glomerular damage? PMID- 12654625 TI - Upregulation and activation of eNOS by resveratrol. PMID- 12654627 TI - Interleukin-13 and leukotrienes: an intersection of pathogenetic schema. PMID- 12654628 TI - CCR3: a key to mast cell phenotypic and functional diversity? PMID- 12654629 TI - Leukotrienes mediate murine bronchopulmonary hyperreactivity, inflammation, and part of mucosal metaplasia and tissue injury induced by recombinant murine interleukin-13. AB - Interleukin (IL)-13 induces bronchopulmonary hyperreactivity (BHR), eosinophilic inflammation, and mucus accumulation in the murine airways. To investigate the potential role of leukotrienes (LT) in mediating these effects, we studied the ability of IL-13 to induce the expression of 5-lipoxygenase (5-LO), we compared the effects of IL-13 and of various leukotrienes on different biological parameters and the interference by the 5-LO inhibitor zileuton (orally, 50 mg/kg, 3 times a day for 3 days), and by some antagonists. The cysteinyl (Cys)-LTs LTC4, LTD4, LTE4, and LTB4, (1 microg/d for 3 d, instilled intratracheally) induced BHR, cell recruitment, fibroblast growth, and mucus production and release into the airways. After the intratracheal instillation of recombinant murine (rm) IL 13, Cys-LT increased in the bronchoalveolar lavage fluid (BALF) at 15 min, followed by lower amounts at 3-6 h. Zileuton inhibited LT production in the BALF, eosinophil and neutrophil sequestration in the lungs, and their passage into the BALF. Zileuton and the Cys-LT-receptor antagonist (ra) LY171883 or MK-571, or the LTB4-ra PH-163 (at 3-10, 5-15, and 10 mg/kg, respectively, administered intratracheally), inhibited BHR by recombinant murine IL-13. Airways mucus after recombinant murine IL-13-challenge was reduced by zileuton and by LY171883, MK 571, and PH-163. LT also induced the vascular endothelium remodelling and collagen deposition. Overall, our results demonstrate the major involvement of LT in the effects of IL-13 on the lung. PMID- 12654630 TI - CC chemokine receptor 3 mobilizes to the surface of human mast cells and potentiates immunoglobulin E-dependent generation of interleukin 13. AB - Eotaxins-1, -2, and -3 mediate the recruitment of blood-borne eosinophils to allergically inflamed tissues by binding CC chemokine receptor (CCR) 3. Mast cells (MCs) are resident tissue cells that also express CCR3. In the present study, we demonstrate that human (h) MCs in nasal polyps and cultured cord blood derived hMCs express CCR3 not only on their surfaces but also in their secretory granules. Activation of hMCs mediated by the high-affinity Fc receptor for immunoglobulin (Ig)E (Fc epsilon RI) increased the surface presentation of CCR3 within 1 h, with a parallel decrease in intracellular CCR3 as determined by flow cytometry on saponin-permeabilized hMCs. Recombinant eotaxin-1 alone did not induce histamine release or cytokine generation, and did not significantly augment IgE-dependent histamine release by interleukin (IL)-4-primed hMCs. Nevertheless, stimulation of hMCs with eotaxin-1 2 h after Fc epsilon RI cross linkage (concomitantly with maximal surface CCR3 expression) increased Fc epsilon RI-dependent IL-13 generation by hMCs, compared with their replicates stimulated simultaneously with both agonists. Thus, hMCs may store CCR3 and rapidly mobilize it to their surface with IgE-dependent activation, providing a novel potential mechanism for enhanced hMC effector function, including IL-13 production. PMID- 12654631 TI - In vivo transglutaminase type 1 expression in normal lung, preinvasive bronchial lesions, and lung cancer. AB - Transglutaminase type 1 (TGase 1) is a member of a class of enzymes that catalyze the cross-linking of proteins, a characteristic feature of epidermal differentiation and squamous metaplasia. The role of TGase 1 has been extensively studied in epidermis but not in the lung. Using a polyclonal anti-TGase 1 antibody prepared in our laboratory (TGase-lac), TGase 1 expression in normal bronchial epithelium, bronchial preinvasive lesions, and lung cancer was characterized. The specificity of the antibody was confirmed by the presence in squamous differentiated bronchial cells of specific 106-kD and 92-kD bands by Western blotting. In addition, immunohistochemistry displayed a recognized pattern of labeling in both normal and tumor cells beneath the cytoplasmic membrane and within the cytosol. TGase 1 was shown to be expressed by cells from bronchial epithelium and bronchial preinvasive lesions, strongly expressed in most non-small-cell lung cancer tumor cells and in apoptotic bodies, but weakly expressed in small-cell lung cancer. The distribution of TGase 1 mRNA correlated with the immunohistochemical profile. These observations suggest that TGase 1 expression is a normal feature of bronchial epithelium and is linked to the process of squamous differentiation occurring in preinvasive lesions. Its role in lung cancer remains to be clarified. PMID- 12654632 TI - Mechanical stress increases RhoA activation in airway smooth muscle cells. AB - Cultured airway smooth muscle cells subjected to cyclic strain respond with increased cytoskeletal organization and contractility resembling effects described with RhoA activation. To test the hypothesis that strain increases cell cytoskeletal organization through RhoA, cells were subjected to strain in the presence of known activators or inhibitors of RhoA. Ten percent cyclic deformational strain (serum-free conditions) increased F-actin staining (152% over control), and this effect was enhanced by serum or lysophosphatidic acid (180%), but decreased (68%) with Clostridium botulinum toxin inhibition of RhoA or with the Rho kinase inhibitor Y27632 (67%). When cells expressing the dominant negative N17-RhoA isoform were subjected to strain, F-actin staining was disorganized and cells failed to elongate or migrate relative to strain direction. When cells expressing a green fluorescent protein (GFP)-RhoA fusion protein were subjected to strain, GFP showed up to 25% greater cell membrane staining than control cells. Finally, strain caused a 4-fold increase in RhoA activation (Rhotekin binding assay), and a 3-fold increase myosin phosphatase phosphorylation that was inhibited by Y27632. We conclude that mechanical stress activates RhoA, an event that may increase airway smooth muscle contractility. PMID- 12654633 TI - Hyperoxia impairs antibacterial function of macrophages through effects on actin. AB - Oxidative stress may impair alveolar macrophage function in patients with inflammatory lung diseases or those exposed to high concentrations of oxygen. We investigated putative mechanisms of injury to macrophages by oxidative stress, using RAW 264.7 cells exposed to 95% oxygen for 48 h. Hyperoxia-exposed macrophages were less able to phagocytose and kill Klebsiella pneumoniae than normoxic controls, despite increased production of nitric oxide, a free radical important in pathogen killing. Exposure of macrophages to hyperoxia had marked effects on the actin cytoskeleton, including increased actin polymerization, loss of cortical actin, formation of stress fibers, de novo synthesis of actin, and actin oxidation. Hyperoxia induced changes in cell morphology, with increased cell size and pseudopod formation. Exposure of macrophages to jasplakinolide, an agent that increases actin polymerization, also impaired their ability to phagocytose Klebsiella. Alveolar macrophages isolated from mice exposed to 100% oxygen for 84 h also demonstrated impaired phagocytic function, as well as similar effects on the actin cytoskeleton and cell morphology to macrophages exposed to hyperoxia in vitro. We conclude that oxidative stress in vitro and in vivo impairs macrophage antibacterial function through effects on actin. PMID- 12654634 TI - Increased susceptibility to RSV infection by exposure to inhaled diesel engine emissions. AB - Although epidemiologic data strongly suggest a role for inhaled environmental pollutants in modulating the susceptibility to respiratory infection in humans, the underlying cellular and molecular mechanisms have not been well studied in experimental systems. The current study assessed the impact of inhaled diesel engine emissions (DEE) on the host response in vivo to a common pediatric respiratory pathogen, respiratory syncytial virus (RSV). Using a relatively resistant mouse model of RSV infection, prior exposure to either 30 microg/m3 particulate matter (PM) or 1,000 microg/m3 PM of inhaled DEE (6 h/d for seven consecutive days) increased lung inflammation to RSV infection as compared with air-exposed RSV-infected C57Bl/6 mice. Inflammatory cells in bronchoalveolar lavage fluid were increased in a dose-dependent manner with regard to the level of DEE exposure, concomitant with increased levels of inflammatory mediators. Lung histology analysis indicated pronounced peribronchial and peribronchiolar inflammation concordant with the level of DEE exposure during infection. Mucous cell metaplasia was markedly increased in the airway epithelium of DEE-exposed mice following RSV infection. Interestingly, both airway and alveolar host defense and immunomodulatory proteins were attenuated during RSV infection by prior DEE exposure. DEE-induced changes in inflammatory and lung epithelial responses to infection were associated with increased RSV gene expression in the lungs following DEE exposure. These findings are consistent with the concept that DEE exposure modulates the lung host defense to respiratory viral infections and may alter the susceptibility to respiratory infections leading to increased lung disease. PMID- 12654635 TI - Binding of interleukin-8 to heparan sulfate and chondroitin sulfate in lung tissue. AB - Interleukin (IL)-8, a member of the CXC chemokine family, is a potent neutrophil chemotactic factor. Mechanisms that regulate the activity of chemokines in tissue are not clear. The goal of this study was to determine whether IL-8 glycosaminoglycan interactions are responsible for the binding of IL-8 in lung tissue. Experiments were performed with a quantitative tissue-binding assay to measure the amount of 125I-IL-8 binding and an in situ tissue-binding assay to characterize the location of IL-8 binding in lung tissue. Confocal microscopy demonstrated IL-8 binding to specific anatomic locations such as cell surfaces and extracellular matrix that were enriched with heparan sulfate and chondroitin sulfate. Removal of heparan sulfate or chondroitin sulfate from lung tissue significantly decreased the binding of 125I-IL-8. Two forms of IL-8 with single amino acid mutations in the glycosaminoglycan-binding domain showed decreased binding. In addition, studies with normal and monomeric IL-8 showed that dimerization increased the binding of 125I-IL-8 in lung tissue. These findings suggest that IL-8-glycosaminoglycan interactions determine the location where IL 8 binds in lung tissue and provides a site for the dimerization of IL-8, which increases the local concentration of IL-8 in the lungs. PMID- 12654636 TI - Expression of the high-affinity choline transporter, CHT1, in the rat trachea. AB - The rate limiting step in neuronal acetylcholine (ACh) synthesis is the uptake of choline by the high-affinity choline transporter (CHT1). Here, we investigated the distribution of CHT1 in the rat trachea. CHT1-mRNA was detected by reverse transcriptase-polymerase chain reaction in trachea without epithelium, abraded tracheal mucosa, and in epithelial cells obtained by laser-assisted cell-picking. Accordingly, CHT1-mRNA could also be detected in tracheal epithelial cells by in situ hybridization. Recently obtained polyclonal rabbit and guinea-pig antisera against a synthetic peptide corresponding to amino acid residues 29-40 of the rat CHT1 sequence localized CHT1 protein in combination with antisera against the vesicular acetylcholine transporter in cholinergic fibers innervating tracheal glands and the tracheal muscle. In case of the tracheal epithelium, CHT1 was restricted to the apical membrane of the ciliated cells, as demonstrated by confocal laser scanning and electron microscopy using an affinity-purified CHT1 antiserum. The close apposition of CHT1 to reported sites of localization of choline acetyltransferase in these cells is strongly in favor of ACh synthesis being fueled by choline uptake via CHT1 after release and breakdown of ACh at the luminal surface. Accordingly, cholinergic regulation of tracheal epithelial function is governed by local release and recycling of ACh by ciliated cells. PMID- 12654637 TI - Pulmonary vascular permeability and ischemic injury in gelsolin-deficient mice. AB - Gelsolin is a potent actin filament regulatory protein that controls cytoskeletal assembly and disassembly. Because cellular gelsolin deficiency leads to pronounced actin stress fiber formation and defective chemotaxis, and similar cytoskeletal remodeling results in endothelial barrier dysfunction, we hypothesized that gelsolin deficient mice would exhibit increased vascular permeability. To test this hypothesis, we compared baseline lung lavage (BAL) protein concentration, wet/dry weight ratio, and osmotic reflection coefficient for albumin (sigma alb) in gelsolin-deficient (gsn-/-) and C57BL/6 (wild-type) mice. In addition, we assessed lung permeability in response to ischemia by evaluating BAL protein concentration after 4, 8, or 24 h of left pulmonary arterial (LPA) occlusion, and lung wet/dry weight ratio and histology after 24 h of LPA occlusion, in gsn-/- and wild-type animals, as compared with control and sham-operated mice. Baseline measurements revealed that BAL protein concentration was 18-fold higher in gsn-/- than in wild-type mice, whereas sigma alb averaged 0.62 + 0.15 in wild-type, as compared with 0.31 + 0.05 in gsn-/- animals, indicating that gelsolin deficiency caused increased pulmonary vascular permeability. Ischemia increased lung permeability (BAL protein and lung wet/dry weight) in both wild-type and gsn-/- mice. However, whereas the fold-increase in BAL protein concentration was less in gsn-/- mice (2- to 4-fold) as compared with wild-type (22- to 34-fold), the duration of ischemia-induced permeability changes was prolonged. Lung wet/dry weight and gross histology following ischemia were comparable in wild-type and gsn-/- animals. These data suggest that gelsolin significantly contributes to maintenance of vascular barrier function in the lung. PMID- 12654638 TI - Secretory component is cleaved by neutrophil serine proteinases but its epithelial production is increased by neutrophils through NF-kappa B- and p38 mitogen-activated protein kinase-dependent mechanisms. AB - We previously showed that expression of polymeric immunoglobulin receptor (pIgR)/secretory component (SC), the epithelial receptor assuming transport of polymeric IgA in mucosal secretions, is strongly decreased in severe chronic obstructive pulmonary disease. Here, we evaluated in vitro the effects of polymorphonuclear neutrophil (PMN) mediators on pIgR/SC. On polyacrylamide gel electrophoresis analysis, soluble SC was rapidly cleaved by supernatants from phorbol-myristate-acetate-activated PMN, through a serine proteinase activity. Moreover, purified PMN serine proteinases also cleaved SC. Similarly, polymeric IgA was rapidly cleaved in monomers by neutrophil elastase, whereas secretory immunoglobulin A was relatively resistant to neutrophil elastase. Surface pIgR on human bronchial epithelial cells was also cleaved by serine proteinases, as shown by immunofluorescence. In contrast, pIgR/SC production by cultured epithelial cells (quantified by enzyme-linked immunosorbent assay) was significantly increased by supernatants from interleukin-8/formylmethionylleucylphenylalanine activated PMN (122.6 +/- 17.3 versus 70.9 +/- 9 ng/mg protein, P < 0.01). Upregulation of pIgR/SC production by bronchial epithelial cells was abolished by nuclear factor kappa B- and p38 mitogen-activated protein kinase (MAPK) inhibitors. Moreover, supernatants from interleukin 8/formylmethionylleucylphenylalanine-activated PMN induced the phosphorylation of I kappa B-alpha and p38 MAPK in epithelial cells, independently of serine proteinases. Thus, PMN serine proteinases cleave pIgR/SC, whereas activated PMN induce an increased pIgR/SC expression through epithelial activation of nuclear factor kappa B and p38 MAPK pathways. PMID- 12654639 TI - The acute respiratory distress syndrome: a role for transforming growth factor beta 1. AB - The acute respiratory distress syndrome (ARDS) remains a major cause of morbidity and mortality. Enhanced fibrosis and elevated procollagen III levels have been linked to increased mortality. We hypothesized that transforming growth factor (TGF)-beta 1 may play an important role in ARDS, given its role in stimulating fibrosis. Using reverse transcriptase in situ polymerase chain reaction (RT in situ PCR) and immunohistochemistry, we analyzed lung tissue from four fibroproliferative ARDS cases and control subjects. We also compared active TGF beta 1 levels in the bronchoalveolar lavage (BAL) fluid of 13 de novo ARDS cases, and 7 normal control subjects. RT in situ PCR showed TGF-beta 1 mRNA expression in fibroproliferative ARDS cases. Immunohistochemistry confirmed protein expression in these samples. Controls were negative for both techniques. In the newly enrolled ARDS cases, TGF-beta 1 levels, as measured by luciferase assay, were elevated in the 11 of 13 samples, averaging 98 +/- 40 pg/mg protein. Controls had no detectable TGF-beta 1 activity. These data suggest that activation of TGF-beta 1 may be important in the early phases of acute lung injury in addition to driving fibroproliferation. These data may lead to new therapeutic approaches in ARDS through more targeted inhibition of fibrosis. PMID- 12654640 TI - Tissue inhibitor of metalloproteinase-3 downregulation in lymphangioleiomyomatosis: potential consequence of abnormal serum response factor expression. AB - Pulmonary lymphangioleiomyomatosis (LAM) is characterized by abnormal smooth muscle-like cell proliferation leading to tissue destruction and cyst formation. We demonstrate that serum response factor (SRF), a critical smooth muscle transcription factor, is overexpressed in LAM cells. To determine whether abnormal SRF levels might have a pathogenic role in LAM, we transfected SRF into mouse lung fibroblasts and performed a cDNA array analysis. High SRF level upregulated the expression of matrix metalloproteinase (MMP)-2 and MMP-14, two MMPs previously shown to be increased in LAM. In addition, SRF down-regulated tissue inhibitor of metalloproteinase (TIMP)-3, one of their inhibitors. TIMP-3 inhibition was further confirmed by reverse transcriptase/polymerase chain reaction, immunoblotting, and immunostaining of human lung fibroblasts transfected with SRF fused to DsRed2 (a red variant of green fluorescent protein). To determine the in vivo significance of our findings, we immunostained 12 LAM cases for TIMP-3. In eight of them, TIMP-3 was ubiquitously present in normal lung parenchyma, but it was absent in LAM lesions. In the remaining cases, including two out of five normal control lungs, the antibody immunoreacted exclusively with elastin, probably due to suboptimal tissue processing. Because timp-3-null mice develop spontaneous emphysema, our findings suggest that SRF mediated TIMP-3 inhibition might contribute to the tissue damage seen in LAM. PMID- 12654641 TI - Platelets promote eosinophil adhesion of patients with asthma to endothelium under flow conditions. AB - During the late-phase asthmatic response, eosinophils migrate to the bronchial tissue and cause severe damage. In this study we compared in vivo primed eosinophils from patients with allergic asthma with eosinophils from healthy control subjects in their adhesion behavior to tumor necrosis factor-alpha activated endothelium under flow conditions (0.8 dyn/cm2). More eosinophils from patients with asthma adhered to activated endothelium, compared with cells from healthy control subjects (1,237 +/- 126 versus 887 +/- 94 cells/mm2, respectively). In the presence of blocking antibodies directed against very late antigen-4 and E-selectin, the residual binding of the cells of individuals with allergic asthma was significantly higher than that of the healthy control subjects (353 +/- 64 versus 123 +/- 31 cells/mm2, respectively, P < 0.02). In addition, secondary tethering or formation of clusters of the eosinophils of patients with allergic asthma was significantly increased compared with the healthy control subjects (cluster indices 1.8 +/- 0.3 versus 0.8 +/- 0.2, respectively, P < 0.05). Because patient cells showed an enhanced interaction with platelets during the perfusions, the role of P-selectin on platelets was investigated. Blocking antibodies directed against P-selectin reduced the enhanced binding and clustering of eosinophils of patients with allergic asthma. We conclude that P-selectin-bearing platelets contribute to secondary tethering processes of eosinophils to activated endothelium. Therefore, platelets might play an important role in the chronic inflammatory processes of these patients. PMID- 12654642 TI - Surfactant protein A decreases nitric oxide production by macrophages in a tumor necrosis factor-alpha-dependent mechanism. AB - Surfactant protein A (SP-A) modulates the lung defense system through regulation of cytokines and nitric oxide (NO) production by alveolar macrophages (AMs). Whether SP-A upregulates or downregulates production of proinflammatory cytokines and NO is controversial. This study demonstrates the molecular mechanism(s) by which SP-A suppresses NO production by activated murine AMs. NO production by interferon-gamma (IFN-gamma) and IFN-gamma plus Mycobacterium avium-stimulated AMs was mediated through tumor necrosis factor-alpha (TNF-alpha) production, as addition of neutralizing anti-TNF-alpha antibodies during AMs stimulation resulted in reduced NO production. SP-A suppressed NO production by activated AMs by inhibiting TNF-alpha production. The maximum inhibitory effect of SP-A on NO production was observed at 20 microg/ml of SP-A concentration. Furthermore, SP-A inhibited activation of nuclear factor-kappa B, a transcription factor required for induction of TNF-alpha and inducible NO synthase genes. These findings suggest that SP-A suppresses NO production by activated AMs by inhibiting TNF alpha secretion and nuclear factor-kappa B activation. This study also highlights the importance of SP-A levels in the lung, as changes in SP-A levels may modulate the local lung defense system. PMID- 12654643 TI - Pseudomonas aeruginosa elastase degrades surfactant proteins A and D. AB - Both in vitro and in vivo studies provide evidence that surfactant protein (SP)-A and SP-D have an important role in the innate immune response to Pseudomonas aeruginosa. In preliminary experiments characterizing the binding of SP-A to this bacteria, we observed the appearance of apparent degradation products of SP-A, and therefore we hypothesized that P. aeruginosa produces an enzyme that degrades SP-A. Incubation of SP-A with P. aeruginosa organisms from several clinical isolates resulted in concentration- and temperature-dependent degradation of SP-A that was inhibited by a metalloproteinase inhibitor, phosphoramidon. The degradative enzyme was purified by anion exchange chromatography and identified by ion trap mass spectroscopy as Pseudomonas elastase, which was shown to directly degrade SP-A and SP-D. Incubation of P. aeruginosa or purified elastase with cell-free bronchoalveolar lavage (BAL) resulted in degradation of SP-A. Furthermore, SP-A degradation fragments were detectable in BAL from lung transplant patients with cystic fibrosis. We speculate that degradation of SP-A and SP-D is a virulence mechanism in the pathogenesis of chronic P. aeruginosa infection. PMID- 12654644 TI - In vivo pharmacodynamics of antifungal drugs in treatment of candidiasis. PMID- 12654645 TI - In vivo pharmacodynamics of HMR 3270, a glucan synthase inhibitor, in a murine candidiasis model. AB - In vivo pharmacokinetic/pharmacodynamic characterization for numerous antibacterial compounds has had a significant impact upon optimal dosing regimen design and the development of in vivo susceptibility breakpoints. More recently, similar characterization has been undertaken for antifungal drug classes. Very little is known of these pharmacodynamic relationships for the new echinocandin class of compounds. We utilized a neutropenic murine model of disseminated candidiasis to describe the time course antifungal activity of HMR 3270, a new glucan synthase inhibitor. Single-dose in vivo time kill studies with four 16 fold escalating doses demonstrated concentration-dependent killing when drug levels in serum were more than four times the MIC. Postantifungal effects were dose dependent, ranging from 8 to 80 h duration. Multiple dosing regimen studies utilized six total doses, four dosing intervals, and a treatment duration of 6 days. Shortening the dosing interval from every 144 h (q144h) to q36h resulted in a fourfold rise in the dose necessary to achieve a net fungistatic effect. The peak/MIC ratio most strongly correlated with treatment outcomes (peak/MIC ratio, R(2) = 98%; ratio of the area under the concentration-time curve from 0 to 24 h to the MIC, R(2) = 79%, percentage of time above the MIC, R(2) = 61%). Studies were also conducted with five additional Candida albicans isolates to determine if a similar peak/MIC ratio was associated with efficacy. In vivo concentration dependent killing was similarly observed in studies with each of the additional isolates. The peak/MIC ratio necessary to produce efficacy was relatively similar among the strains studied (P = 0.42). The peak/MIC ratio (mean +/- standard deviation) necessary to achieve a fungistatic effect was 3.72 +/- 1.84, and the ratio necessary to achieve maximal killing was near 10. PMID- 12654646 TI - In vivo pharmacodynamics of a new triazole, ravuconazole, in a murine candidiasis model. AB - In vivo studies have characterized the pharmacodynamic characteristics of the triazole fluconazole. These investigations demonstrated that the ratio of the area under the concentration-time curve from 0 to 24 h to the MIC (24-h AUC/MIC ratio) is the critical pharmacokinetic/pharmacodynamic (PK/PD) parameter associated with treatment efficacy. Further analysis demonstrated that a fluconazole 24-h AUC/MIC ratio of 20 to 25 was predictive of treatment success in both experimental models and clinical trials. We used a neutropenic murine model of disseminated Candida albicans infection to similarly characterize the time course activity of the new triazole ravuconazole. The PK/PD parameters (percent time above the MIC, AUC/MIC ratio, and peak level in serum/MIC ratio) were correlated with in vivo efficacy, as measured by organism number in kidney cultures after 24 and 72 h of therapy. Ravuconazole kinetics and protein binding were performed in neutropenic infected mice. Peak/dose and AUC/dose values ranged from 0.03 to 0.04 and 0.30 to 0.34, respectively. Serum elimination half-life ranged from 3.9 to 4.8 h. Protein binding was 95.8%. Single-dose postantifungal effect studies demonstrated prolonged suppression of organism regrowth after serum ravuconazole levels had fallen below the MIC. Treatment efficacies with the five dosing intervals studied were similar, supporting the argument for the AUC/MIC ratio as the PK/PD parameter predictive of efficacy. Nonlinear regression analysis also suggested that the AUC/MIC ratio was strongly predictive of treatment outcomes (AUC/MIC ratio, R(2) = 91%; peak/MIC ratio, R(2) = 85%; percent time above the MIC, R(2) = 47 to 65%). Similar studies were conducted with seven additional C. albicans isolates with various ravuconazole susceptibilities (MIC, 0.016 to 0.12 micro g/ml) to determine if a similar 24-h AUC/MIC ratio was associated with efficacy. The ravuconazole free-drug AUC/MIC ratios were similar for all of the organisms studied (10 to 36; mean +/- SD = 20.3 +/- 8.2; P = 0.43). These free-drug AUC/MIC ratios are similar to those observed for fluconazole in this model. PMID- 12654647 TI - Sublethal injury and resuscitation of Candida albicans after amphotericin B treatment. AB - Amphotericin B treatment was previously shown to inhibit Candida albicans reproduction and reduce the fluorescence of vitality-specific dyes without causing a corresponding increase in the fluorescence of the mortality-specific dyes bis-(1,3-dibutylbarbituric acid)trimethine oxonol and SYBR Green I. In the present study, we have confirmed these results and have shown that the numbers of CFU are reduced by 99.9% by treatment with 0.5 micro g of amphotericin B per ml for 10 h at 35 degrees C. This reduction was not due to fungal cell death. First, the level of reduction of the tetrazolium salt 2,3-bis(2-methoxy-4-nitro-5 sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide increased in the presence of concentrations of amphotericin B that caused greater than 90% reductions in the numbers of CFU. Second, fungal cells treated with amphotericin B at a concentration of 0.5 micro g/ml were resuscitated by further incubation at 22 degrees C for 15 h in the continued presence of amphotericin B. Third, recovery of the ability to replicate was prevented by sequential treatment with 20 micro g of miconazole per ml, which also increased the fluorescence of mortality-specific dyes to near the maximal levels achieved with 0.9 micro g of amphotericin B per ml. Sequential treatment with fluconazole and flucytosine did not increase the levels of staining with the mortality-specific dyes. Itraconazole was less effective than ketoconazole, which was less effective than miconazole. The practice of equating the loss of the capacity of C. albicans to form colonies with fungal cell death may give incorrect results in assays with amphotericin B, and the results of assays with caution with other antifungal agents that are lipophilic or that possess significant postantifungal effects may need to be interpreted. PMID- 12654648 TI - Drug resistance is not directly affected by mating type locus zygosity in Candida albicans. AB - Recently, evidence was presented that in a collection of fluconazole-resistant strains of Candida albicans there was a much higher proportion of homozygotes for the mating type locus (MTL) than in a collection of fluconazole-sensitive isolates, suggesting the possibility that when cells become MTL homozygous they acquire intrinsic drug resistance. To investigate this possibility, an opposite strategy was employed. First, drug susceptibility was measured in a collection of isolates selected for MTL homozygosity. The majority of these isolates had not been exposed to antifungal drugs. Second, the level of drug susceptibility was compared between spontaneously generated MTL-homozygous progeny and their MTL heterozygous parent strains which had not been exposed to antifungal drugs. The results demonstrate that naturally occurring MTL-homozygous strains are not intrinsically more drug resistant, supporting the hypotheses that either the higher incidence of MTL homozygosity previously demonstrated among fluconazole resistant isolates involved associated homozygosity of a drug resistance gene linked to the MTL locus, or that MTL-homozygous strains may be better at developing drug resistance upon exposure to the drug than MTL-heterozygous strains. Furthermore, the results demonstrate that a switch by an MTL-homozygous strain from the white to opaque phenotype, the latter functioning as the facilitator of mating, does not notably alter drug susceptibility. PMID- 12654649 TI - Genetic basis for differential activities of fluconazole and voriconazole against Candida krusei. AB - Invasive infections caused by Candida krusei are a significant concern because this organism is intrinsically resistant to fluconazole. Voriconazole is more active than fluconazole against C. krusei in vitro. One mechanism of fluconazole resistance in C. krusei is diminished sensitivity of the target enzyme, cytochrome P450 sterol 14alpha-demethylase (CYP51), to inhibition by this drug. We investigated the interactions of fluconazole and voriconazole with the CYP51s of C. krusei (ckCYP51) and fluconazole-susceptible Candida albicans (caCYP51). We found that voriconazole was a more potent inhibitor of both ckCYP51 and caCYP51 in cell extracts than was fluconazole. Also, the ckCYP51 was less sensitive to inhibition by both drugs than was caCYP51. These results were confirmed by expressing the CYP51 genes from C. krusei and C. albicans in Saccharomyces cerevisiae and determining the susceptibility of the transformants to voriconazole and fluconazole. We constructed homology models of the CYP51s of C. albicans and C. krusei based on the crystal structure of CYP51 from Mycobacterium tuberculosis. These models predicted that voriconazole is a more potent inhibitor of both caCYP51 and ckCYP51 than is fluconazole, because the extra methyl group of voriconazole results in a stronger hydrophobic interaction with the aromatic amino acids in the substrate binding site and more extensive filling of this site. Although there are multiple differences in the predicted amino acid sequence of caCYP51 and ckCYP51, the models of the two enzymes were quite similar and the mechanism for the relative resistance of ckCYP51 to the azoles was not apparent. PMID- 12654650 TI - Genome-wide expression profile analysis reveals coordinately regulated genes associated with stepwise acquisition of azole resistance in Candida albicans clinical isolates. AB - Candida albicans is an opportunistic human fungal pathogen and a causative agent of oropharyngeal candidiasis (OPC), the most frequent opportunistic infection among patients with AIDS. Fluconazole and other azole antifungal agents have proven effective in the management of OPC; however, with increased use of these agents treatment failures have occurred. Such failures have been associated with the emergence of azole-resistant strains of C. albicans. In the present study we examined changes in the genome-wide gene expression profile of a series of C. albicans clinical isolates representing the stepwise acquisition of azole resistance. In addition to genes previously associated with azole resistance, we identified many genes whose differential expression was for the first time associated with this phenotype. Furthermore, the expression of these genes was correlated with that of the known resistance genes CDR1, CDR2, and CaMDR1. Genes coordinately regulated with the up-regulation of CDR1 and CDR2 included the up regulation of GPX1 and RTA3 and the down-regulation of EBP1. Genes coordinately regulated with the up-regulation of CaMDR1 included the up-regulation of IFD1, IFD4, IFD5, IFD7, GRP2, DPP1, CRD2, and INO1 and the down-regulation of FET34, OPI3, and IPF1222. Several of these appeared to be coordinately regulated with both the CDR genes and CaMDR1. Many of these genes are involved in the oxidative stress response, suggesting that reduced susceptibility to oxidative damage may contribute to azole resistance. Further evaluation of the role these genes and their respective gene products play in azole antifungal resistance is warranted. PMID- 12654651 TI - Isolation of three Mycobacterium ulcerans strains resistant to rifampin after experimental chemotherapy of mice. AB - By use of a murine model for Buruli ulcer, Mycobacterium ulcerans was found to be susceptible to rifampin, with the MIC being 0.5 to 1 micro g/ml. Three mutants were isolated after rifampin monotherapy. Two were resistant to rifampin at 8 micro g/ml, and one was resistant to rifampin at 32 micro g/ml. The mutants harbored Ser416Phe mutations and His420Tyr mutations in the rpoB gene, and these mutations have also been found to be responsible for rifampin resistance in the leprosy and tubercle bacilli. The results indicate that while rifampin may be active against M. ulcerans, it should never be used as monotherapy in humans. PMID- 12654652 TI - In vivo antiretroviral activity of stampidine in chronically feline immunodeficiency virus-infected cats. AB - Here we report the antiretroviral activity of the experimental nucleoside reverse transcriptase inhibitor (NRTI) compound stampidine in cats chronically infected with feline immunodeficiency virus (FIV). Notably, a single oral bolus dose of 50 or 100 mg of stampidine per kg resulted in a transient >/=1-log decrease in the FIV load of circulating peripheral blood mononuclear cells in five of six FIV infected cats and no side effects. A 4-week stampidine treatment course with twice-daily administration of hard gelatin capsules containing 25 to 100 mg of stampidine per kg was also very well tolerated by cats at cumulative dose levels as high as 8.4 g/kg and exhibited a dose-dependent antiretroviral effect. One of three cats treated at the 25-mg/kg dose level, three of three cats treated at the 50-mg/kg dose level, and three of three cats treated at the 100-mg/kg dose level (but none of three control cats treated with placebo pills) showed a therapeutic response, as evidenced by a >/=1-log reduction in the FIV load in peripheral blood mononuclear cells within 2 weeks. The previously documented in vitro and in vivo antiretroviral activity of stampidine against primary clinical human immunodeficiency virus type 1 isolates with genotypic and/or phenotypic NRTI resistance, together with its favorable animal toxicity profile, pharmacokinetics, and in vivo antiretroviral activity in FIV-infected cats, warrants further development of this promising new NRTI compound. PMID- 12654653 TI - Single nucleotide polymorphisms in genes associated with isoniazid resistance in Mycobacterium tuberculosis. AB - Isoniazid (INH) is a central component of drug regimens used worldwide to treat tuberculosis. Previous studies have identified resistance-associated mutations in katG, inhA, kasA, ndh, and the oxyR-ahpC intergenic region. DNA microarray-based experiments have shown that INH induces several genes in Mycobacterium tuberculosis that encode proteins physiologically relevant to the drug's mode of action. To gain further insight into the molecular genetic basis of INH resistance, 20 genes implicated in INH resistance were sequenced for INH resistance-associated mutations. Thirty-eight INH-monoresistant clinical isolates and 86 INH-susceptible isolates of M. tuberculosis were obtained from the Texas Department of Health and the Houston Tuberculosis Initiative. Epidemiologic independence was established for all isolates by IS6110 restriction fragment length polymorphism analysis. Susceptible isolates were matched with resistant isolates by molecular genetic group and IS6110 profiles. Spoligotyping was done with isolates with five or fewer IS6110 copies. A major genetic group was established on the basis of the polymorphisms in katG codon 463 and gyrA codon 95. MICs were determined by the E-test. Semiquantitative catalase assays were performed with isolates with mutations in the katG gene. When the 20 genes were sequenced, it was found that 17 (44.7%) INH-resistant isolates had a single locus, resistance-associated mutation in the katG, mabA, or Rv1772 gene. Seventeen (44.7%) INH-resistant isolates had resistance-associated mutations in two or more genes, and 76% of all INH-resistant isolates had a mutation in the katG gene. Mutations were also identified in the fadE24, Rv1592c, Rv1772, Rv0340, and iniBAC genes, recently shown by DNA-based microarray experiments to be upregulated in response to INH. In general, the MICs were higher for isolates with mutations in katG and the isolates had reduced catalase activities. The results show that a variety of single nucleotide polymorphisms in multiple genes are found exclusively in INH-resistant clinical isolates. These genes either are involved in mycolic acid biosynthesis or are overexpressed as a response to the buildup or cellular toxicity of INH. PMID- 12654654 TI - Role of nutrient limitation and stationary-phase existence in Klebsiella pneumoniae biofilm resistance to ampicillin and ciprofloxacin. AB - Biofilms formed by Klebsiella pneumoniae resisted killing during prolonged exposure to ampicillin or ciprofloxacin even though these agents have been shown to penetrate bacterial aggregates. Bacteria dispersed from biofilms into medium quickly regained most of their susceptibility. Experiments with free-floating bacteria showed that stationary-phase bacteria were protected from killing by either antibiotic, especially when the test was performed in medium lacking carbon and nitrogen sources. These results suggested that the antibiotic tolerance of biofilm bacteria could be explained by nutrient limitation in the biofilm leading to stationary-phase existence of at least some of the cells in the biofilm. This mechanism was supported by experimental characterization of nutrient availability and growth status in biofilms. The average specific growth rate of bacteria in biofilms was only 0.032 h(-1) compared to the specific growth rate of planktonic bacteria of 0.59 h(-1) measured in the same medium. Glucose did not penetrate all the way through the biofilm, and oxygen was shown to penetrate only into the upper 100 micro m. The specific catalase activity was elevated in biofilm bacteria to a level similar to that of stationary-phase planktonic cells. Transmission electron microscopy revealed that bacteria were affected by ampicillin near the periphery of the biofilm but were not affected in the interior. Taken together, these results indicate that K. pneumoniae in this system experience nutrient limitation locally within the biofilm, leading to zones in which the bacteria enter stationary phase and are growing slowly or not at all. In these inactive regions, bacteria are less susceptible to killing by antibiotics. PMID- 12654655 TI - High-efficiency generation of antibiotic-resistant strains of Streptococcus pneumoniae by PCR and transformation. AB - We designed a method by which to generate antibiotic-resistant strains of Streptococcus pneumoniae at frequencies 4 orders of magnitude greater than the spontaneous mutation rate. The method is based on the natural ability of this organism to be genetically transformed with PCR products carrying sequences homologous to its chromosome. The genes encoding the targets of ciprofloxacin (parC, encoding the ParC subunit of DNA topoisomerase IV), rifampin (rpoB, encoding the beta subunit of RNA polymerase), and streptomycin (rpsL, encoding the S12 ribosomal protein) from susceptible laboratory strain R6 were amplified by PCR and used to transform the same strain. Resistant mutants were obtained with a frequency of 10(-4) to 10(-5), depending on the fidelity of the DNA polymerase used for PCR amplifications. Ciprofloxacin-resistant mutants, for which the MICs were four-to eightfold higher than that for R6, carried a single mutation of a residue in the quinolone resistance-determining region: S79 (change to A, F, or Y) or D83 (change to N or V). Rifampin-resistant strains, for which the MICs were at least 133-fold higher than that for R6, contained a single mutation within cluster I of rpoB: S482 (change to P), Q486 (change to L), D489 (change to V), or H499 (change to L or Y). Streptomycin-resistant mutants, for which the MICs were at least 64-fold higher than that for R6, carried a mutation at either K56 (change to I, R, or T) or K101 (change to E). PCR products obtained from the mutants were able to transform R6 to resistance with high efficiency (>10(4)). This method could be used to efficiently obtain resistant mutants for any drug whose target is known. PMID- 12654656 TI - Vancomycin treatment failure associated with heterogeneous vancomycin intermediate Staphylococcus aureus in a patient with endocarditis and in the rabbit model of endocarditis. AB - Heterogeneous resistance to vancomycin is thought to precede emergence of intermediate susceptibility to vancomycin in Staphylococcus aureus, but the clinical significance of heterogeneous resistance is unknown. Paired S. aureus isolates from a patient with endocarditis who relapsed after vancomycin treatment were tested for heterogeneous resistance to vancomycin. The pretreatment and the relapse clinical isolates (strains SF1 and SF2, respectively) were genotyped by pulsed-field gel electrophoresis. Susceptibility to vancomycin was assessed by the broth dilution method, population analysis, and time-kill studies and in the rabbit model of endocarditis. Strains SF1 and SF2 had similar genotypes, and the vancomycin MICs for the strains were /= 128 micro g/ml) and susceptible to fluconazole (0.5 < MIC < 2 micro g/ml), four became flucytosine-fluconazole cross resistant when both antifungals were used simultaneously. Fluconazole resistance occurred only in the presence of high flucytosine concentrations, and the higher the fluconazole concentration used, the greater the flucytosine concentration necessary to trigger the cross resistance. When the flucytosine- and fluconazole-resistant cells were grown in the presence of fluconazole alone, the cells reversed to fluconazole susceptibility. Genetic analyses of the progeny from crosses between resistant and sensitive isolates showed that resistance to flucytosine was derived from a recessive mutation in a single gene, whereas cross-resistance to fluconazole seemed to vary like a quantitative trait. We further demonstrated that the four clinical isolates were susceptible to 5-fluorouracil and that cytosine deaminase activity was unaffected. Kinetic transport studies with [(14)C]flucytosine showed that flucytosine resistance was due to a defect in the purine-cytosine permease. Our hypothesis was that extracellular flucytosine would subsequently behave as a competitive inhibitor of fluconazole uptake transport. Finally, in vitro selection of spontaneous and induced mutants indicated that such a cross resistance mechanism could also affect other Candida species, including C. albicans, C. tropicalis, and C. glabrata. This is the first report of a putative fluconazole uptake transporter in Candida species and of a possible resistance mechanism associated with a deficiency in the uptake of this drug. PMID- 12654659 TI - Integron-containing IncU R plasmids pRAS1 and pAr-32 from the fish pathogen Aeromonas salmonicida. AB - A 45-kb R plasmid, pRAS1, that confers resistance to tetracyclines, trimethoprim, and sulfonamides was isolated in 1989 from an atypical strain of the fish pathogen Aeromonas salmonicida. This plasmid could be transferred by conjugation to Escherichia coli with a high degree of efficiency (frequency, 0.48). The following year pRAS1 was isolated from A. salmonicida subsp. salmonicida in the same area. Incompatibility group U plasmid pRAS1 contained a drug resistance determining region of 12 kb consisting of a class 1 integron similar to In4 of Tn1696 but with a dfrA16 gene cassette inserted. Close to IS6100 at the right end of Tn4 was a truncated Tn1721. Restriction enzyme analysis showed that R plasmid pAr-32, isolated from A. salmonicida in Japan in 1970, had the same backbone structure as pRAS1, while the drug resistance-determining region contained a complex class 1 integron with an aadA2 cassette; the chloramphenicol resistance gene catA2, as in In6 of pSa; and a duplicate of the 3' conserved segment of the integron. PMID- 12654660 TI - Engineering anthracycline biosynthesis toward angucyclines. AB - The biosynthesis pathways of two anthracyclines, nogalamycin and aclacinomycin, were directed toward angucyclines by using an angucycline-specific cyclase, pgaF, isolated from a silent antibiotic biosynthesis gene cluster. Addition of pgaF to a gene cassette that harbored the early biosynthesis genes of nogalamycin resulted in the production of two known angucyclinone metabolites, rabelomycin and its precursor, UWM6. Substrate flexibility of pgaF was demonstrated by replacement of the nogalamycin minimal polyketide synthase genes in the gene cassette with the equivalent aclacinomycin genes together with aknE2 and aknF, which specify the unusual propionate starter unit in aclacinomycin biosynthesis. This modification led to the production of a novel angucyclinone, MM2002, in which the expected ethyl side chain was incorporated into the fourth ring. PMID- 12654661 TI - Imipenem resistance in a Salmonella clinical strain due to plasmid-mediated class A carbapenemase KPC-2. AB - A Salmonella enterica serotype Cubana isolate exhibiting resistance to most beta lactam antibiotics, including oxyimino-cephalosporins and imipenem, was isolated from a 4-year-old boy with gastroenteritis in Maryland. beta-Lactam resistance was mediated by a conjugative plasmid that encoded KPC-2, a class A carbapenemase previously found in a Klebsiella pneumoniae isolate from the Maryland area as well. Sequence analysis of the flanking regions indicated a potential association of bla(KPC-2) with mobile structures. PMID- 12654662 TI - Use of genetically engineered phage to deliver antimicrobial agents to bacteria: an alternative therapy for treatment of bacterial infections. AB - The emergence and increasing prevalence of multidrug-resistant bacterial pathogens emphasizes the need for new and innovative antimicrobial strategies. Lytic phages, which kill their host following amplification and release of progeny phage into the environment, may offer an alternative strategy for combating bacterial infections. In this study, however, we describe the use of a nonlytic phage to specifically target and deliver DNA encoding bactericidal proteins to bacteria. To test the concept of using phage as a lethal-agent delivery vehicle, we used the M13 phagemid system and the addiction toxins Gef and ChpBK. Phage delivery of lethal-agent phagemids reduced target bacterial numbers by several orders of magnitude in vitro and in a bacteremic mouse model of infection. Given the powerful genetic engineering tools available and the present knowledge in phage biology, this technology may have potential use in antimicrobial therapies and DNA vaccine development. PMID- 12654663 TI - Bactericidal activities of methoxyfluoroquinolones gatifloxacin and moxifloxacin against aerobic and anaerobic respiratory pathogens in serum. AB - Gatifloxacin (Bristol-Myers Squibb) and moxifloxacin (Bayer) are new methoxyfluoroquinolones with broad-spectrum activity against aerobic and anaerobic pathogens of the respiratory tract. In this investigation, we analyzed the bactericidal activity in serum over time of these antimicrobials against three aerobic (Streptococcus pneumoniae, Haemophilus influenzae, and Staphylococcus aureus) and four anaerobic (Peptostreptococcus micros, Peptostreptococcus magnus, Fusobacterium nucleatum, and Prevotella melaninogenica) bacteria associated with respiratory tract infections. Serum samples were obtained from 11 healthy male subjects following a single 400-mg oral dose of gatifloxacin and moxifloxacin. These samples were collected prior to and at 2, 6, 12, and 24 h after the dose of each drug. Gatifloxacin exhibited bactericidal activity for a median of 12 h against Streptococcus pneumoniae (MIC = 0.5 micro g/ml), Peptostreptococcus micros (MIC = 0.25 micro g/ml), and F. nucleatum (MIC = 0.5 micro g/ml) and 24 h against H. influenzae (MIC = 0.03 micro g/ml), Staphylococcus aureus (MIC = 0.125 micro g/ml), Peptostreptococcus magnus (MIC = 0.125 micro g/ml), and Prevotella melaninogenica (MIC = 0.5 micro g/ml). Moxifloxacin exhibited bactericidal activity for a median of 24 h against Streptococcus pneumoniae (MIC = 0.125 micro g/ml), H. influenzae (MIC = 0.015 micro g/ml), Staphylococcus aureus (MIC = 0.06 micro g/ml), F. nucleatum (MIC = 0.5 micro g/ml), Prevotella melaninogenica (MIC =0.5 micro g/ml), Peptostreptococcus magnus (MIC = 0.125 micro g/ml), and Peptostreptococcus micros (MIC = 0.25 micro g/ml). The results from this pharmacodynamic study suggest that these fluoroquinolones would have prolonged killing activity against these organisms in vivo and may have clinical utility in the treatment of mixed aerobic anaerobic respiratory tract infections. PMID- 12654664 TI - Rifampin followed by ceftriaxone for experimental meningitis decreases lipoteichoic acid concentrations in cerebrospinal fluid and reduces neuronal damage in comparison to ceftriaxone alone. AB - Rifampin (RIF) releases smaller quantities of lipoteichoic acids (LTAs) from Streptococcus pneumoniae than ceftriaxone (CRO). Due to the rapid development of resistance, RIF cannot be used as a single agent for therapy of bacterial meningitis. For this reason, we compared the effect of treatment with RIF followed by treatment with CRO (RIF-CRO) or the effect of treatment with clindamycin (CLI) followed by treatment with CRO (CLI-CRO) to that of CRO alone on the concentrations of LTAs and teichoic acids in vitro. The effects of RIF-CRO on LTA concentrations in cerebrospinal fluid (CSF) and on neuronal injury were investigated in a rabbit model of S. pneumoniae meningitis. In vitro, bacterial titers were effectively reduced by CRO, RIF-CRO, and CLI-CRO when each drug was used at 10 micro g/ml. The levels of release of LTAs after the initiation of therapy were lower in RIF-CRO- and CLI-CRO-treated cultures than in cultures treated with CRO alone (P < 0.05 from 3 to 12 h after initiation of treatment). Similarly, in rabbits, the increase in the amount of LTAs in CSF was lower in RIF CRO-treated animals than in CRO-treated animals (P = 0.02). The density of dentate apoptotic granular cells was lower after RIF-CRO therapy than after CRO therapy (medians, 58.4 and 145.6/mm(2), respectively; 25th quartiles, 36.3 and 81.7/mm(2), respectively; 75th quartiles, 100.7 and 152.3/mm(2), respectively; P = 0.03). Therefore, initiation of therapy with a protein synthesis-inhibiting antibacterial and continuation of therapy with a combination that includes a beta lactam may be a strategy to decrease neuronal injury in bacterial meningitis. PMID- 12654665 TI - Daptomycin pharmacokinetics and safety following administration of escalating doses once daily to healthy subjects. AB - The purpose of this paper is to establish the pharmacokinetics and safety of escalating, once-daily doses of daptomycin, a novel lipopeptide antibiotic active against gram-positive pathogens, including those resistant to methicillin and vancomycin. This phase 1, multiple-dose, double-blind study involved 24 healthy subjects in three dose cohorts (4, 6, and 8 mg/kg of body weight) who were randomized to receive daptomycin or the control at a 3:1 ratio and administered the study medication by a 30-min intravenous infusion every 24 h for 7 to 14 days. Daptomycin pharmacokinetics was assessed by blood and urine sampling. Safety and tolerability were evaluated by monitoring adverse events (AEs) and laboratory parameters. Daptomycin pharmacokinetics was linear through 6 mg/kg, with a slight ( approximately 20%) nonlinearity in the area under the curve and trough concentration at the highest dose studied (8 mg/kg). The pharmacokinetic parameters measured on the median day of the study period, (day 7) were half-life ( approximately 9 h), volume of distribution ( approximately 0.1 liters/kg), systemic clearance ( approximately 8.2 ml/h/kg), and percentage of the drug excreted intact in urine from 0 to 24 h ( approximately 54%). Daptomycin protein binding (mean amount bound, 91.7%) was independent of the drug concentration. No gender effect was observed. All subjects who received daptomycin completed the study. The frequencies and distributions of treatment-emergent AEs were similar for the subjects who received daptomycin and the control subjects. There were no serious AEs and no pattern of dose-related events. The pharmacokinetics of once daily administration of daptomycin was linear through 6 mg/kg. For all three doses, plasma daptomycin concentrations were consistent and predictable throughout the dosing interval. Daptomycin was well tolerated when it was administered once daily at a dose as high as 8 mg/kg for 14 days. PMID- 12654667 TI - Phase I safety and pharmacokinetic trials of 1263W94, a novel oral anti-human cytomegalovirus agent, in healthy and human immunodeficiency virus-infected subjects. AB - 1263W94 [maribavir; 5,6-dichloro-2-(isopropylamino)-1, beta-L-ribofuranosyl-1-H benzimidazole], a novel benzimidazole compound, has been demonstrated to potently and selectively inhibit human cytomegalovirus replication in vitro and to have favorable safety profiles in animal species. Two phase I trials evaluated the safety and pharmacokinetics of escalating single doses of 1263W94 in 13 healthy subjects (dose, 50 to 1,600 mg) and 17 human immunodeficiency virus (HIV) infected subjects (dose, 100 to 1,600 mg). No severe safety concerns were observed in the evaluation of adverse events, vital signs, electrocardiograms, and clinical laboratory tests following administration of a single dose of 1263W94. The most frequently reported adverse events in both populations were taste disturbance (80%) and headache (53%). 1263W94 was rapidly absorbed following oral administration, with peak concentrations in plasma (C(max)) occurring 1 to 3 h after dosing. The increases in the C(max) of 1263W94 and the area under the concentration-time curve from time zero to infinity (AUC(0- infinity )) for 1263W94 were dose dependent; C(max) increased slightly less than proportionally to the dose, and AUC(0- infinity ) increased slightly more than proportionally to the dose. 1263W94 was rapidly eliminated, with a mean half-life in plasma of 3 to 5 h; the half-life was independent of the dose level. Less than 2% of the 1263W94 dose administered was eliminated unchanged in urine. The principal metabolite of 1263W94 was 4469W94 (which is derived by N-dealkylation of 1263W94 via CYP3A4), which accounted for 30 to 40% of the dose in urine. Greater than 98% of the 1263W94 in plasma is bound to proteins, and the extent of binding appears to be constant over the dose range of 200 to 1,600 mg. In the trial with HIV-infected subjects, consumption of a high-fat meal decreased the 1263W94 AUC(0- infinity ) and C(max) in plasma by approximately 30%. PMID- 12654666 TI - Activities of atazanavir (BMS-232632) against a large panel of human immunodeficiency virus type 1 clinical isolates resistant to one or more approved protease inhibitors. AB - To evaluate the cross-resistance profile of the human immunodeficiency virus type 1 protease inhibitor (PI) atazanavir (BMS-232632), a panel of 551 clinical isolates exhibiting a wide array of PI resistance profiles and a variety of genotypic patterns were assayed for susceptibility to atazanavir and six other PIs: amprenavir, indinavir, lopinavir, nelfinavir, ritonavir, and saquinavir. In general, reductions in atazanavir susceptibility in vitro required several amino acid changes and were relatively modest in degree, and susceptibility was retained among isolates resistant to one or two of the currently approved PIs. There was a clear trend toward loss of susceptibility to atazanavir, as isolates exhibited increasing levels of cross-resistance to multiple PIs. Atazanavir appeared to have a distinct resistance profile relative to each of the other six PIs tested based on susceptibility comparisons against this panel of resistant isolates. Analysis of the genotypic profiles of 943 PI-susceptible and -resistant clinical isolates identified a strong correlation between the presence of amino acid changes at specific residues (10I/V/F, 20R/M/I, 24I, 33I/F/V, 36I/L/V, 46I/L, 48V, 54V/L, 63P, 71V/T/I, 73C/S/T/A, 82A/F/S/T, 84V, and 90M) and decreased susceptibility to atazanavir. While no single substitution or combination of substitutions was predictive of atazanavir resistance (change, >3.0-fold), the presence of at least five of these substitutions correlated strongly with loss of atazanavir susceptibility. Mutations associated with reduced susceptibility to each of the other six PIs were also determined. PMID- 12654668 TI - Chromosomal aadD2 encodes an aminoglycoside nucleotidyltransferase in Bacillus clausii. AB - Bacillus clausii SIN is one of the four strains of B. clausii composing a probiotic administered to humans for the prevention of gastrointestinal side effects due to oral antibiotic therapy. The strain is resistant to kanamycin, tobramycin, and amikacin. A gene conferring aminoglycoside resistance was cloned into Escherichia coli and sequenced. The gene, called aadD2, encoding a putative 246-amino acid protein, shared 47% identity with ant(4')-Ia from Staphylococcus aureus, which encodes an aminoglycoside 4'-O-nucleotidyltransferase. Phosphocellulose paper-binding assays indicated that the gene product was responsible for nucleotidylation of kanamycin, tobramycin, and amikacin. The aadD2 gene was detected by DNA-DNA hybridization in the three other strains of the probiotic mixture and in the reference strain B. clausii DSM8716, although it did not confer resistance in these strains. Mutations in the sequence of the putative promoter for aadD2 from B. clausii SIN resulted in higher identity with consensus promoter sequences and may account for aminoglycoside resistance in that strain. The aadD2 gene was chromosomally located in all strains and was not transferable by conjugation. These data indicate that chromosomal aadD2 is specific to B. clausii. PMID- 12654669 TI - Molecular determination of point mutation haplotypes in the dihydrofolate reductase and dihydropteroate synthase of Plasmodium falciparum in three districts of northern Tanzania. AB - The antimalarial combination of sulfadoxine and pyrimethamine (SP) was introduced as first-line treatment for uncomplicated malaria in Tanzania during 2001 following 18 years of second-line use. The genetic determinants of in vitro resistance to the two drugs individually are shown to be point mutations at seven sites in the dihydrofolate reductase gene (dhfr) conferring resistance to pyrimethamine and five sites in the dihydropteroate synthase (dhps) gene conferring resistance to sulfadoxine. Different combinations of mutations within each gene confer differing degrees of insensitivity, but information about the frequency with which allelic haplotypes occur has been lacking because of the complicating effects of multiple infection. Here we used a novel high-throughput sequence-specific oligonucleotide probe-based approach to examine the present resistance status of three Plasmodium falciparum populations in northern Tanzania. By using surveys of asymptomatic infections and screening for the presence of all known point mutations in dhfr and dhps genes, we showed that just five dhfr and three dhps allelic haplotypes are present. High frequencies of both triple-mutant dhfr and double-mutant dhps mutant alleles were found in addition to significant interregional heterogeneity in allele frequency. In vivo studies have shown that the cooccurrence of three dhfr mutations and two dhps mutations in an infection prior to treatment is statistically predictive of treatment failure. We have combined data for both loci to determine the frequency of two locus genotypes. The triple-dhfr/double-dhps genotype is present in all three regions with frequencies ranging between 30 and 63%, indicating that treatment failure rates are likely to be high. PMID- 12654670 TI - Pharmacokinetics and efficacy of linezolid in a gerbil model of Streptococcus pneumoniae-induced acute otitis media. AB - The oxazolidinone linezolid represents a new antibacterial class of potential benefit in managing multidrug-resistant gram-positive infections, including those caused by Streptococcus pneumoniae. In a gerbil model of acute otitis media (AOM) induced by either penicillin-resistant S. pneumoniae (PRSP; amoxicillin MIC = 8 micro g/ml, linezolid MIC = 1 micro g/ml) or penicillin-susceptible S. pneumoniae (PSSP; amoxicillin MIC = 0.015 micro g/ml, linezolid MIC = 1 micro g/ml), we explored the plasma and ear fluid levels of linezolid required to demonstrate efficacy. Threshold pathogen doses required to induce bilateral AOM (1,500 CFU/ear with PRSP; 30 CFU/ear with PSSP) were administered to gerbils by intrabullar injection on day 0. At peak infection ( approximately 10(6) to 10(7) CFU/ear flush; day 2 for PRSP-AOM and day 3 for PSSP-AOM), twice-a-day oral doses of linezolid, amoxicillin, or vehicle were administered over 4.5 days prior to collection and assay of middle ear effluents for S. pneumoniae content. Linezolid doses of >/=10 mg/kg of body weight induced significant cure rates of >/=72% versus both PRSP and PSSP infections, whereas amoxicillin at MIC of >/=42%, a C(max)/MIC ratio of >/=3.1, and a (24-h area under the curve)/MIC ratio of >/=30 h. Application of this model will be useful in defining preclinical pharmacodynamic relationships of novel antibiotics necessary to cure S. pneumoniae-induced AOM. PMID- 12654671 TI - Stability of colistin and colistin methanesulfonate in aqueous media and plasma as determined by high-performance liquid chromatography. AB - The stabilities of colistin and colistin methanesulfonate (CMS) in different aqueous media were studied by specific high-performance liquid chromatography (HPLC) methods. Colistin was stable in water at 4 and 37 degrees C for up to 60 days and 120 h, respectively. However, degradation was observed when colistin was stored in isotonic phosphate buffer (0.067 M, pH 7.4) and human plasma at 37 degrees C. The stability of CMS from three different sources in water was explored by strong-anion-exchange (SAX) HPLC for CMS and by measuring the concentrations of colistin formed from the hydrolysis of CMS. The peaks of CMS in SAX HPLC disappeared almost completely after 12 h at 37 degrees C, but appeared to remain intact for up to 2 days at 4 degrees C. Over the same period, there was no formation of colistin at 4 degrees C. In water, phosphate buffer, and plasma, there was rapid formation of colistin within 24 to 48 h at 37 degrees C from the three sources of CMS. The hydrolysis products were assumed to be a complex mixture of many different sulfomethyl derivatives, including colistin. The stability of a fourth source of CMS in Mueller-Hinton broth examined during 30 min at 37 degrees C revealed no formation of colistin. Along with previous microbiological studies, this suggested that different sulfomethyl CMSs possess intrinsic antibacterial activity. These results will be helpful for understanding the pharmacokinetics and pharmacodynamics of colistin and CMS in humans and animals. PMID- 12654672 TI - MexXY-OprM efflux pump is necessary for a adaptive resistance of Pseudomonas aeruginosa to aminoglycosides. AB - Exposure of Pseudomonas aeruginosa to aminoglycosides frequently selects for recalcitrant subpopulations exhibiting an unstable, "adaptive" resistance to these antibiotics. In this study, we investigated the implication in the phenomenon of MexXY-OprM, an active efflux system known to export aminoglycosides in P. aeruginosa. Immunoblotting experiments demonstrated that the transporter MexY, but not the outer membrane pore OprM, was overproduced during the post-drug exposure adaptation period in wild-type strain PAO1. Furthermore, MexY production was dependent upon the degree of bacterial exposure to gentamicin (drug concentration). In contrast to parental strain PAO1, mutants defective in MexXY or in OprM were unable to develop adaptive resistance. Altogether, these results indicate that the resistance process requires the rapid production of MexXY and the interaction of these proteins with the constitutively produced component OprM. PMID- 12654673 TI - In vitro antifungal activity of Micafungin (FK463) against dimorphic fungi: comparison of yeast-like and mycelial forms. AB - The characteristics of in vitro micafungin (FK463) antifungal activity against six species of dimorphic fungi were investigated in accordance with the NCCLS M27 A microdilution methods. MICs of micafungin, amphotericin B, itraconazole, and fluconazole for Histoplasma capsulatum var. capsulatum, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Penicillium marneffei, and Sporothrix schenckii were determined both for the yeast-like form and mycelial form. Coccidioides immitis was tested only in its mycelial form. We have clearly demonstrated that the in vitro activity of micafungin depends considerably on the growth form of dimorphic fungi. Micafungin exhibited potent activity against the mycelial forms of H. capsulatum, B. dermatitidis, and C. immitis (MIC range, 0.0078 to 0.0625 micro g/ml), while it was very weakly active against their yeast-like forms (MIC range, 32 to >64 micro g/ml). Micafungin was also more active against the mycelial forms than the yeast-like forms of Paracoccidioides brasiliensis, Penicillium marneffei, and S. schenckii. The MICs of amphotericin B were 2 to 5 dilutions lower for the mycelial forms than for the yeast-like forms of B. dermatitidis and Paracoccidioides brasiliensis. There was no apparent difference in the activity of itraconazole between the two forms. The MICs of fluconazole for the yeast-like forms were generally lower than those for the mycelial forms, and considerably so for B. dermatitidis. These results suggest that the growth form employed in antifungal susceptibility testing of dimorphic fungi can considerably influence the interpretation of results. At present, it cannot be judged whether micafungin has clinical usefulness for dimorphic fungus infections, since for most fungi it remains uncertain which growth form correlates better with therapeutic outcome. However, the results of this study warrant further investigations of micafungin as a therapeutic agent for infections caused by dimorphic fungi. PMID- 12654675 TI - In vitro activities of piperaquine and other 4-aminoquinolines against clinical isolates of Plasmodium falciparum in Cameroon. AB - The spread of chloroquine-resistant Plasmodium falciparum calls for a constant search for new drugs. The in vitro activity of piperaquine, a new Chinese synthetic drug belonging to the bisquinolines, was evaluated in 103 fresh clinical isolates of P. falciparum in Cameroon, Central Africa, and compared with that of other 4-aminoquinoline and Mannich base derivatives and dihydroartemisinin. Piperaquine was highly active (geometric mean 50% inhibitory concentration, 38.9 nmol/liter; range, 7.76 to 78.3 nmol/liter) and equally active (P > 0.05) against the chloroquine-sensitive and the chloroquine-resistant isolates. There was a significant but low correlation of response between chloroquine and piperaquine (r = 0.257, P < 0.05). These results suggest that further development of piperaquine, in combination with dihydroartemisinin, holds promise for use in chloroquine-resistant regions of endemicity. PMID- 12654674 TI - Epidemiology of rifampin ADP-ribosyltransferase (arr-2) and metallo-beta lactamase (blaIMP-4) gene cassettes in class 1 integrons in Acinetobacter strains isolated from blood cultures in 1997 to 2000. AB - We characterized two new gene cassettes in an Acinetobacter isolate: one harbored the metallo-beta-lactamase (IMP-4) gene bla(IMP-4), the other harbored the rifampin ADP-ribosyltransferase (ARR-2) gene arr-2, and both arrayed with the aminoglycoside acetyltransferase [AAC(6')-Ib(7)] gene cassette aacA4 in two separate class 1 integrons. The epidemiology of these gene cassettes in isolates from blood cultures obtained from 1997 to 2000 was studied. Isolates bearing either the bla(IMP-4) or the arr-2 gene cassette or both represented 17.5% (10 of 57) of isolates in 1997, 16.1% (10 of 62) in 1998, 2.5% (1 of 40) in 1999, and 0% (0 of 58) in 2000. These two gene cassettes, probably borne on two separate integrons, were found in at least three genomic DNA groups, with evidence of clonal dissemination in the intensive care unit during 1997 to 1998. Seventeen of the 52 Acinetobacter baumannii (genomic DNA group 2) isolates from 1997 to 2000 harbored intI1, but only one was positive for these gene cassettes, whereas 20 of the 21 intI1-positive isolates of all other genomic DNA groups were positive for either or both of them. Reduced susceptibility to imipenem and rifampin was seen only in isolates harboring the bla(IMP-4) and arr-2 cassettes, respectively. The aminoglycoside phosphotransferase [APH(3')-VIa] gene aph(3')-VIa was detected in all 21 isolates for which the MIC of amikacin was >/=8 micro g/ml, with or without aacA4, whereas aacA4 alone was found in isolates for which the MIC of amikacin was 0.5 to 2 micro g/ml. Significant differences between the 17 intI1 positive and 47 intI1-negative isolates belonging to genomic DNA group 3 from 1997 to 1998 in the MICs of amikacin, gentamicin, imipenem, sulfamethoxazole, and ceftazidime were observed (Mann-Whitney test, P < 0.001 to 0.01). PMID- 12654676 TI - Pharmacokinetics and metabolism of [(14)C]ribavirin in rats and cynomolgus monkeys. AB - Absorption, pharmacokinetics, distribution, metabolism, and excretion of [(14)C]ribavirin were studied in rats (30 mg/kg of body weight) and cynomolgus monkeys (10 mg/kg) after intravenous (i.v.) and oral administration. The oral absorption and bioavailability were 83 and 59%, respectively, in rats and 87 and 55%, respectively, in monkeys. After i.v. administration, the elimination half life (t([1/2])) was 9.9 h in rats and 130 h in monkeys and the total body clearance was 2,600 ml/h/kg in rats and 224 ml/h/kg in monkeys. The apparent volume of distribution was 11.4 liter/kg in rats and 29.4 liter/kg in monkeys. There was extensive distribution of drug-derived radioactivity into red blood cells and extensive metabolism of ribavirin in rats and a lesser degree of metabolism in monkeys. Excretion of total radioactivity in urine from rats accounted for 84% of the i.v. dose and 83% of the oral dose, whereas that from monkeys accounted for 47% of the i.v. dose and 67% of the oral dose. Several metabolites were observed in plasma and urine from both species. The amount of unchanged ribavirin in urine from both species was quite small after either i.v. or oral administration. PMID- 12654677 TI - Time-kill studies of the antianaerobe activity of garenoxacin compared with those of nine other agents. AB - The activities of garenoxacin, ciprofloxacin, levofloxacin, moxifloxacin, trovafloxacin, amoxicillin-clavulanate, piperacillin-tazobactam, imipenem, clindamycin, and metronidazole against 20 anaerobes were tested. At two times the MIC, garenoxacin was bactericidal against 19 of 20 strains after 48 h and against 17 of 20 after 24 h. Other drugs, except clindamycin (which gave lower killing rates), gave killing rates similar to those for garenoxacin. PMID- 12654678 TI - Activities of ABT-773 against Listeria monocytogenes and coryneform bacteria of clinical interest. AB - The in vitro activities of ABT-773 were evaluated against 15 Listeria monocytogenes strains and 196 coryneform bacteria isolated from clinical samples. One hundred percent of the L. monocytogenes strains were inhibited by 32 (Corynebacterium jeikeium), 0.03 and >32 (Corynebacterium minutissimum), >32 and >32 (Corynebacterium pseudodiphtheriticum and Corynebacterium urealyticum), 0.125 and >32 (Corynebacterium striatum), and 0.03 and 0.5 (Rhodococcus equi), respectively. PMID- 12654679 TI - In vitro metronidazole and tinidazole activities against metronidazole-resistant strains of Trichomonas vaginalis. AB - The in vitro activities of tinidazole and metronidazole against Trichomonas vaginalis isolates clinically resistant to metronidazole were compared. Minimal lethal concentrations (MLCs) of tinidazole were significantly lower than MLCs of metronidazole. Increased metronidazole resistance correlated with increased tinidazole resistance. These data support a role for tinidazole in the treatment of trichomoniasis. PMID- 12654681 TI - In vitro synergy of caspofungin and itraconazole against Aspergillus spp.: MIC versus minimal effective concentration end points. AB - Caspofungin and itraconazole were studied alone and in combination against 31 clinical isolates of Aspergillus spp. according to NCCLS M38-P guidelines. MICs and microscopic minimal effective concentrations (MECs) were recorded, and synergy was calculated by using both end points. Synergy or synergy to additivity was found in 30 of 31 isolates by using MIC end points. With MEC end points no synergy was found and indifference was detected in 26 of 31 strains. PMID- 12654680 TI - Activity of and resistance to moxifloxacin in Staphylococcus aureus. AB - Moxifloxacin has enhanced potency against Staphylococcus aureus, lower propensity to select for resistant mutants, and higher bactericidal activity against highly resistant strains than ciprofloxacin. Despite similar activity against purified S. aureus topoisomerase IV and DNA gyrase, it selects for topoisomerase IV mutants, making topoisomerase IV the preferred target in vivo. PMID- 12654683 TI - Multiplex PCR for detection of aminoglycoside resistance genes in enterococci. AB - A multiplex PCR procedure for detecting the aminoglycoside resistance genes aac(6')-Ie-aph(2")-Ia, aph(2")-Ib, aph(2")-Ic, aph(2")-Id, aph(3')-IIIa, and ant(4')-Ia was evaluated and found to determine accurately the presence of these genes in enterococci. PMID- 12654682 TI - Genetic characterization of fluoroquinolone-resistant Streptococcus pneumoniae strains isolated during ciprofloxacin therapy from a patient with bronchiectasis. AB - Five Spain(9V-3) Streptococcus pneumoniae strains were isolated from a patient with bronchiectasis who had received long-term ciprofloxacin therapy. One ciprofloxacin-susceptible strain was isolated before treatment, and four ciprofloxacin-resistant strains were isolated during treatment. The resistant strains were derived from the susceptible strain either by a parC mutation (low level resistance) or by parC and gyrA mutations (high-level resistance). This study shows that ciprofloxacin therapy in a patient colonized by susceptible S. pneumoniae may select fluoroquinolone-resistant mutants. PMID- 12654685 TI - Novel tetracycline resistance determinant from the oral metagenome. AB - A major drawback of most studies on how bacteria become resistant to antibiotics is that they concentrate mainly on bacteria that can be cultivated in the laboratory. In the present study, we cloned part of the oral metagenome and isolated a novel tetracycline resistance gene, tet(37), which inactivates tetracycline. PMID- 12654684 TI - Integron-associated antibiotic resistance in Salmonella enterica serovar typhi from Asia. AB - Eighteen of 25 isolates of Salmonella enterica serovar Typhi were multidrug resistant and contained class 1 integrons with a single cassette, dfrVII or aadA1. The dfrVII-containing integron was likely borne on an IncHI1 plasmid. Salmonella serovar Typhi could become resistant to broad-spectrum cephalosporins by integrating cassettes, such as veb-1, a common cassette in Asia. PMID- 12654686 TI - Molecular characterization and antimicrobial susceptibility of fluoroquinolone resistant or -susceptible Streptococcus pneumoniae from Hong Kong. AB - Fluoroquinolone resistance in Streptococcus pneumoniae isolated from Hong Kong as part of Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin 1999/2000 was found to be due to the spread of the Spain(23F)-1 clone (mainly a Spain(23F)-1-14 variant). All the isolates were multidrug resistant but were susceptible to quinupristin-dalfopristin, linezolid, and telithromycin. The Spain(23F)-1 clone also occurred among antimicrobial susceptible isolates, which suggests that the primary source of this clone may have been Asia rather than Iberia. PMID- 12654687 TI - Activity of posaconazole against Pseudallescheria boydii: in vitro and in vivo assays. AB - Thirty isolates of Pseudallescheria boydii were tested to compare the in vitro activity of posaconazole with those of fluconazole and itraconazole, using NCCLS methods. Posaconazole was evaluated in an immunosuppressed mouse model of disseminated pseudallescheriasis. Posaconazole was more effective than itraconazole and as effective as fluconazole in preventing death and significantly reducing the CFU of P. boydii from tissues. PMID- 12654688 TI - Tissue penetration by ertapenem, a parenteral carbapenem administered once daily, in suction-induced skin blister fluid in healthy young volunteers. AB - The penetration of 1 g of intravenous ertapenem once daily for 3 days in suction induced skin blisters was evaluated. Ten forearm blisters were formed (n = 12) 12 h prior to the last dose. Concentrations of ertapenem in blister fluid exceeded 4 micro g/ml (the MIC at which 90% of the isolates tested are eliminated) for the entire dosing interval. The area under the concentration-time curve for 0 to 24 h ratio of blister fluid to plasma was 61% (90% confidence interval, 56, 65%) suggesting good blister penetration. PMID- 12654689 TI - Antibiotic resistance in bacteria isolated from subgingival plaque in a norwegian population with refractory marginal periodontitis. AB - The prevalence of beta-lactamase-producing bacteria in subgingival plaque from patients with refractory periodontitis in Norway was assessed by the chromogenic nitrocefin method. beta-Lactamase activity was detected in 68% of the patients. Enzyme-producing strains belonged predominantly to the genus Prevotella; others were staphylococci, enteric gram-negative rods, and Bacillus spp. PMID- 12654690 TI - In vitro activity of a new antibiotic, NVP-PDF386 (VRC4887), against Chlamydia pneumoniae. AB - The in vitro activity of NVP-PDF386 (VRC4887), a novel new peptide deformylase inhibitor, and those of levofloxacin and clarithromycin were tested against 21 isolates of Chlamydia pneumoniae. The MIC at which 90% of the isolates were inhibited and the minimal bactericidal concentration at which 90% of the isolates were killed by NVP-PDF386 for all isolates of C. pneumoniae were 0.008 micro g/ml (range, 0.008 to 0.015 micro g/ml) compared to 0.25 and 0.06 micro g/ml for levofloxacin and clarithromycin, respectively. PMID- 12654692 TI - Efficacy of micafungin alone or in combination against systemic murine aspergillosis. AB - We tested the efficacy of micafungin (FK) alone or in combination with other antifungals against systemic murine aspergillosis. FK alone at 10 mg/kg of body weight/dose prolonged survival (P = 0.01) and reduced CFU in the brain and kidney. Combination therapy that used suboptimal FK with amphotericin B or itraconazole prolonged survival. Although no survivors were free of infection, no antagonism was seen. Nikkomycin Z with FK showed significantly greater potency (P < 0.01) than either alone. PMID- 12654691 TI - Trypanocidal and leishmanicidal properties of substitution-containing chalcones. AB - Ten chalcones were synthesized and tested as potential leishmanicidal and trypanocidal agents. All tested compounds caused concentration-dependent inhibition of the in vitro growth of Leishmania braziliensis and Trypanosoma cruzi with no significant toxic effect towards host macrophages. Our results show that the positions of the substituents seem to be critical for their antiprotozoal activities. PMID- 12654693 TI - PCR-oligonucleotide ligation assay for detection of point mutations associated with quinolone resistance in Streptococcus pneumoniae. AB - We have developed a PCR-oligonucleotide ligation assay to rapidly identify base substitutions in topoisomerase genes that are associated with quinolone resistance in clinical isolates of Streptococcus pneumoniae. Thirty-seven strains for which the ciprofloxacin MICs were >/=4 micro g/ml and 16 strains for which the MICs were T in the G protein beta3 subunit gene and 190G-->A in the CC chemokine receptor 2 gene) were significantly associated with hypertension in men and that one polymorphism (-238G-->A in the tumor necrosis factor alpha gene) was significantly associated with hypertension in women. These results suggest that 2 and 1 genes may be susceptibility loci for hypertension in Japanese men and women, respectively, and that genotyping of these polymorphisms may prove informative for prediction of the genetic risk for hypertension. PMID- 12654704 TI - Estrogen depletion increases blood pressure and hypothalamic norepinephrine in middle-aged spontaneously hypertensive rats. AB - In male spontaneously hypertensive rats (SHR) a high NaCl diet increases arterial pressure via a reduction in anterior hypothalamic nucleus norepinephrine release. Young female SHR are relatively well protected from this NaCl-sensitive hypertension, but depletion of both endogenous and dietary estrogens greatly exacerbates NaCl-sensitive hypertension. This study tests the hypothesis that estrogen also protects late middle-aged female SHR from NaCl-sensitive hypertension and that this effect is mediated by an estrogen-related effect on hypothalamic norepinephrine release. Ten-month-old female SHR were ovariectomized and placed on a phytoestrogen-free diet containing either basal or high NaCl. Each rat was implanted with a silastic tube containing 17beta estradiol or vehicle. Three months later, arterial pressure and hypothalamic norepinephrine metabolite levels (MOPEG) were measured. On the basal NaCl diet, estrogen depleted rats displayed increased arterial pressure (12 mm Hg) and decreased anterior hypothalamic nucleus MOPEG (20%). Both effects were reversed by estrogen treatment. In all groups, the high NaCl diet increased arterial pressure by over 35 mm Hg and reduced anterior hypothalamic nucleus MOPEG by >60%. Across all groups, there was a significant inverse correlation between arterial pressure and anterior hypothalamic nucleus MOPEG. These data suggest that both dietary NaCl excess and estrogen depletion raise arterial pressure in middle-aged female SHR by a decreasing hypothalamic norepinephrine. PMID- 12654705 TI - An acyl-CoA synthetase gene family in chromosome 16p12 may contribute to multiple risk factors. AB - We recently reported that genetic polymorphisms of SAH, an acyl-CoA synthetase for fatty acids, might contribute to multiple risk factors, especially hypertriglyceridemia. There are at least 4 members in this SAH gene family, SAH, MACS1, MACS2, and MACS3, and these 4 members are clustered in human Ch16p12. It is possible either that the previously observed associations were due to linkage disequilibrium with truly important polymorphisms in other members of the SAH gene family or that other polymorphisms in this gene family may also influence multiple risk factors. Thus, we performed association studies between genetic polymorphisms in this SAH region and multiple risk factors, using a large cohort representing the general population in Japan. The L513S polymorphism in MACS2 was shown to significantly influence the triglyceride level and the waist-to-hip ratio. The previously observed associations between an SAH polymorphism and the waist-to-hip ratio appear to be due to linkage disequilibrium with the L513S polymorphism. Haplotype analysis indicated that a haplotype defined by the I/D polymorphism of SAH and the L513S polymorphism in MACS2 was highly significantly associated with the triglyceride level. This study confirmed the importance of this chromosomal region in the pathogenesis of hypertriglyceridemia and visceral obesity. PMID- 12654706 TI - Effect of low-dose perindopril/indapamide on albuminuria in diabetes: preterax in albuminuria regression: PREMIER. AB - Microalbuminuria in diabetes is a risk factor for early death and an indicator for aggressive blood pressure (BP) lowering. We compared a combination of 2 mg perindopril/0.625 mg indapamide with enalapril monotherapy on albumin excretion rate (AER) in patients with type 2 diabetes, albuminuria, and hypertension in a 12-month, randomized, double-blind, parallel-group international multicenter study. Four hundred eighty-one patients with type 2 diabetes and hypertension (systolic BP > or =140 mm Hg, <180 mm Hg, diastolic BP <110 mm Hg) were randomly assigned (age 59+/-9 years, 77% previously treated for hypertension). Results from 457 patients (intention-to-treat analysis) were available. After a 4-week placebo period, patients with albuminuria >20 and <500 microg/min were randomly assigned to a combination of 2 mg perindopril/0.625 mg indapamide or to 10 mg daily enalapril. After week 12, doses were adjusted on the basis of BP to a maximum of 8 mg perindopril/2.5 mg indapamide or 40 mg enalapril. The main outcome measures were overnight AER and supine BP. Both treatments reduced BP. Perindopril/indapamide treatment resulted in a statistically significant higher fall in both BP (-3.0 [95% CI -5.6, -0.4], P=0.012; systolic BP -1.5 [95% CI 3.0, -0.1] diastolic BP P=0.019) and AER -42% (95% CI -50%, -33%) versus -27% (95% CI -37%, -16%) with enalapril. The greater AER reduction remained significant after adjustment for mean BP. Adverse events were similar in the 2 groups. Thus, first-line treatment with low-dose combination perindopril/indapamide induces a greater decrease in albuminuria than enalapril, partially independent of BP reduction. A BP-independent effect of the combination may increase renal protection. PMID- 12654708 TI - AT1 receptors in the RVLM mediate pressor responses to emotional stress in rabbits. AB - In this study, we examined the role of angiotensin type 1 (AT1) receptors in the rostral ventrolateral medulla (RVLM) in mediating the pressor action of emotional stress in conscious rabbits. Rabbits were chronically instrumented with guide cannulas for bilateral microinjections into the RVLM and an electrode for measuring renal sympathetic nerve activity (RSNA). Airjet stress evoked increases in arterial pressure, heart rate, and RSNA, which reached a maximum (+9+/-1 mm Hg, +20+/-5 beats/min, and +93+/-17%, respectively) in the first 2 minutes of stress exposure. Then RSNA rapidly returned to prestress values, while arterial pressure and heart rate remained close to the maximal level until the conclusion of the 7-minute airjet exposure. Microinjections of the nonselective angiotensin receptor antagonist sarile (0.5 nmol, n=8) or AT1 receptor antagonists losartan (2 nmol, n=6) or candesartan (0.2 nmol, n=6) into the RVLM did not alter resting cardiovascular parameters. By contrast, the antagonists attenuated the sustained phase (4 to 7 minutes) of the pressor stress response by 55% to 89%. However, only sarile decreased the onset of this response. The antagonists affected neither the stress-induced tachycardia nor the pressor response to glutamate microinjections. Microinfusion of angiotensin II (4 pmol/min, n=8) into the RVLM did not change the pressor response to airjet stress but attenuated tachycardic response by 47%. Microinjections of vehicle did not alter the cardiovascular stress response. Sarile, losartan, and angiotensin II did not affect the sympathoexcitatory response to baroreceptor unloading. These results suggest that AT1 receptors in the RVLM are important in mediating the pressor effects of emotional stress in conscious rabbits. PMID- 12654707 TI - AT1-receptor antagonism improves endothelial function in coronary artery disease by a bradykinin/B2-receptor-dependent mechanism. AB - Impaired flow-dependent, endothelium-mediated vasodilation is an early finding in patients with coronary artery disease (CAD). Experimental and some clinical studies observed that angiotensin type-1 receptor antagonists (AT1A) enhance endothelium-dependent relaxation in CAD. The present study was designed to determine whether AT1A improves flow-dependent dilation (FDD) in patients with CAD and, if so, whether bradykinin and NO are involved. High-resolution ultrasound was used to measure radial artery diameter at rest and during reactive hyperemia, causing endothelium-mediated vasodilation. Twenty patients with CAD were randomly assigned to receive intrabrachial infusion of candesartan (800 microg/min) with and without icatibant, a bradykinin B2-receptor antagonist (90 microg/min; group A) or N-monomethyl-l-arginine (L-NMMA), an NO-synthase inhibitor (7 micromol/min; group B). The AT1A candesartan improved FDD by >40%, an effect that was inhibited by icatibant (group A: control, 7.3+/-0.9; candesartan, 10.3+/-1.1; candesartan+icatibant, 5.0+/-0.5%). Similarly, L-NMMA blunted the beneficial effect of candesartan (group B: control, 6.3+/-0.6; candesartan, 8.9+/-0.6; candesartan+L-NMMA: 4.7+/-0.5%; each P<0.01). The angiotensin type-1 receptor antagonist candesartan improves flow-dependent, endothelium-mediated vasodilation in patients with CAD. This effect is inhibited by either icatibant and or L-NMMA, suggesting that both bradykinin and NO contribute to the vascular effects of AT1-receptor antagonists in this patient population. PMID- 12654710 TI - Reliability of different echocardiographic methods in determining left ventricular mass in healthy men and athletes. PMID- 12654711 TI - Segment of rat chromosome 20 regulates diet-induced augmentations in adiposity, glucose intolerance, and blood pressure. AB - Previous linkage and association studies have suggested that a region of human chromosome 6 containing the tumor necrosis factor (TNF)-alpha gene is involved in the pathogenesis of obesity and obesity-associated hypertension. The aim of the present investigation was to establish whether a segment of rat chromosome 20 (RNO20), which also contains the TNF-alpha gene, determines diet-induced changes in adiposity and blood pressure (BP). The results showed that a transfer of the RNO20 segment from the normotensive Brown Norway (BN) rat onto the background of the spontaneously hypertensive rat (SHR) is associated with a significantly greater increase in adiposity, glucose intolerance, circulating leptin levels, and BP during 12-week, high-fat-diet feeding. In contrast, the transfer is not associated with significant changes in these variables during 12-week, normal diet feeding. In addition, sequencing of the TNF-alpha gene revealed differences between SHR and BN in the 5'- and 3'-regulatory regions of the gene. Subsequent analyses of TNF-alpha gene expression in fat, muscle, and liver, however, did not provide support for the functional involvement of these differences. In summary, the investigated RNO20 segment contains 1 or more gene variants that affect adiposity, glucose tolerance, serum leptin levels, and BP, but only when the animals are exposed to a particular environment, ie, high-fat-diet feeding. Further studies are needed to identify genes mediating these effects. Considering current changes in our lifestyle involving an increased calorie and fat intake, we believe that gene-environment interactions, such as those described here, play an important role in the current epidemic of obesity and obesity-associated hypertension. PMID- 12654712 TI - Transactivation of KDR/Flk-1 by the B2 receptor induces tube formation in human coronary endothelial cells. AB - Endothelial cells (ECs) are the critical cellular element responsible for postnatal angiogenesis. Vascular endothelial growth factor (VEGF) stimulates angiogenesis via the activation of kinase insert domain-containing receptor/fetal liver kinase-1 (KDR/Flk-1) in ECs. In addition, transactivation of KDR/Flk-1 by the bradykinin (BK) B2 receptor induces the activation of endothelial nitric oxide synthase (eNOS). These findings indicate that the precise role of BK in angiogenesis is likely to be more complex than initially thought, and it questions the importance of BK in angiogenic processes. Therefore, we examined whether transactivation by BK induced tube formation. We developed an in vitro model of human coronary artery EC (HCEC) tube formation on a matrix gel. We demonstrated that BK dose-dependently induced tube formation. Although a lower concentration of BK and VEGF did not separately induce tube formation, the formation was induced by a combination of lower concentrations of BK and VEGF, suggesting that VEGF and BK had a synergistic effect. The effect was blocked by a B2 receptor antagonist (HOE140) and specific inhibitors of VEGF receptor tyrosine kinases (Tki) and NOS. In addition, BK induced tyrosine phosphorylation of the KDR/Flk-1 receptor, as did VEGF itself. The transactivation was also blocked by HOE140 and Tki. Our results showed that, in HCECs, stimulation of the B2 receptor leads to the transactivation of KDR/Flk-1, as well as to eNOS activation, which induces tube formation. To our knowledge, this is a novel mechanism in which transactivation of KDR/Flk-1 by a G protein-coupled receptor, B2 receptor, may be a potent signal for tube formation. PMID- 12654713 TI - Is altered adrenal steroid biosynthesis a key intermediate phenotype in hypertension? AB - Approximately 10% of patients with hypertension have a high ratio of aldosterone to renin, but the reason for this and the relationships among low-renin essential hypertension, elevation of the ratio, and true primary aldosteronism are unclear. We have previously reported that a polymorphism of the gene (C-to-T conversion at position -344) encoding aldosterone synthase is associated with hypertension, particularly in patients with a high ratio. However, the most consistent association with this variant is a relative impairment of adrenal 11beta hydroxylation. In this review, we propose that altered conversion of deoxycortisol to cortisol leads to a subtle, chronic increase in adrenocortrophin drive to the adrenal cortex, with eventual development of hyperplasia. In combination with other genetic or environmental factors (such as dietary sodium intake), we suggest that this might be responsible for the long-term development of a resetting of the aldosterone response to angiotensin II, giving rise to the phenotype of hypertension with a raised ratio. In some subjects, this may progress further to true primary aldosteronism with a dominant adrenal nodule. Thus, there may be a genetically influenced continuum from hypertension with a normal ratio, through hypertension with a raised ratio, and primary aldosteronism. PMID- 12654714 TI - Racial differences in the prevalence of hypertensive retinopathy. AB - Few population-based data support the hypothesis that hypertensive retinopathy is more common in African Americans than in whites. We examined racial differences in the prevalence of and risk factors for hypertensive retinopathy in a population-based sample of 1860 African Americans and 7874 white persons, aged 49 to 73 years, without diabetes. Retinal photographs were taken of one randomly selected eye and evaluated for the presence of retinopathy (flame and blot-shaped retinal hemorrhages, microaneurysms, and soft exudates) according to standardized protocols by graders masked to participant characteristics. The prevalence of retinopathy was 2 times higher in African Americans than in whites (7.7% versus 4.1%, age- and gender-adjusted odds ratio [OR] 2.03, 95% confidence intervals [CI] 1.65, 2.49). After controlling for 6-year mean arterial blood pressure, use of antihypertensive medications and left ventricular hypertrophy by ECG criteria, the excess prevalence of retinopathy in African Americans was reduced by 40% (adjusted OR 1.61, 95% CI 1.26, 2.06). Further adjustment for other vascular risk factors, common carotid artery intima-media thickness, and serum creatinine levels reduced the excess prevalence in African Americans by another 13% (adjusted OR 1.48, 95% CI 1.08, 2.03). We conclude that hypertensive retinopathy is twice as frequent in African Americans compared with whites without diabetes and that the excess prevalence of retinopathy in African Americans is associated with blood pressure and severity of hypertension. PMID- 12654716 TI - Hypothesis: ACE2 modulates blood pressure in the mammalian organism. PMID- 12654715 TI - Cardiac hypertrophy and microvascular deficit in kinin B2 receptor knockout mice. AB - Experimental and clinical evidence suggests kinin involvement in adaptive myocardial growth. Kinins are growth-inhibitory to cardiomyocytes. Knockout of kinin B2 receptor (B2R) signaling causes dilated and failing cardiomyopathy in 129/J mice, and a 9-bp deletion polymorphism of human B2R is associated with reduced receptor expression and exaggerated left ventricular growth response to physical stress. We reasoned that genetic background and aging may significantly influence the impact of B2R mutation on cardiac phenotype. The theory was challenged in C57BL/6 mice, a strain that naturally differs from the 129/J strain, carrying 1 instead of 2 renin genes. C57BL/6 B2R knockouts (B2R-KO) showed higher blood pressure and heart rate levels (P<0.05) compared with wild type controls (WT) at all ages examined. At 12 months, left ventricular contractility and diastolic function were mildly altered (P<0.05) and histological and morphological analyses revealed ventricular hypertrophy and cardiomyocyte enlargement in B2R-KO (P<0.01). Reparative fibrosis was enhanced by 208% and capillary density reduced by 38% (P<0.01). Functional and structural alterations induced by B2R deletion in C57BL/6 mice were less severe than those reported previously in the 129/J strain. We conclude that interaction of B2R signaling with other genetic determinants influences aging-related changes in myocardial structure and function. These findings may help us understand the role of kinins in the development of cardiac failure. PMID- 12654717 TI - Maternal-fetal flow, negative events, and preeclampsia: role of ACE I/D polymorphism. AB - The risk for an adverse pregnancy outcome is markedly higher in women with history of preeclampsia. This may stem from impaired placentation in early gestation and from high impedance to flow in uteroplacental circulation. The renin-angiotensin system is one of the mediators of the remodeling of spiral arteries throughout pregnancy. The D allele of the Insertion/Deletion (I/D) polymorphism in the ACE gene has been associated with higher ACE activity, accounting for 47% of the total phenotypic variance of serum enzyme levels. To investigate whether the ACE I/D polymorphism affects maternal uteroplacental and fetal umbilical circulation and the pregnancy outcome in women with a history of preeclampsia, 106 women underwent Doppler examination of uterine arteries resistance index and umbilical artery pulsatility index at the 16th, 20th, and 24th weeks of gestation and were genotyped for the I/D polymorphism. This study found a difference in genotype distribution (P=0.0002) and allele frequency (P<0.0001) between women with and those without preeclampsia recurrence and fetal growth restriction as well as an association (P=0.0007) between DD genotype and risk of recurrent preeclampsia or fetal growth restriction. At the 16th, 20th, and 24th weeks, uterine artery resistance indexes were significantly lower in II, higher in DD, and intermediate in ID genotype carriers, whereas the umbilical artery pulsatility index values were significantly higher in the DD group in comparison to ID and II genotypes. The current study shows that the ACE I/D polymorphism affects uteroplacental and umbilical flows and the recurrence of an adverse pregnancy outcome in women with history of preeclampsia. PMID- 12654718 TI - Novel multilocus measure of linkage disequilibrium to estimate past effective population size. AB - Linkage disequilibrium (LD) between densely spaced, polymorphic genetic markers in humans and other species contains information about historical population size. Inferring past population size is of interest both from an evolutionary perspective (e.g., testing the "out of Africa" hypothesis of human evolution) and to improve models for mapping of disease and quantitative trait genes. We propose a novel multilocus measure of LD, the chromosome segment homozygosity (CSH). CSH is defined for a specific chromosome segment, up to the full length of the chromosome. In computer simulations CSH was generally less variable than the r(2) measure of LD, and variability of CSH decreased as the number of markers in the chromosome segment was increased. The essence and utility of our novel measure is that CSH over long distances reflects recent effective population size (N), whereas CSH over small distances reflects the effective size in the more distant past. We illustrate the utility of CSH by calculating CSH from human and dairy cattle SNP and microsatellite marker data, and predicting N at various times in the past for each species. Results indicated an exponentially increasing N in humans and a declining N in dairy cattle. CSH is a valuable statistic for inferring population histories from haplotype data, and has implications for mapping of disease loci. PMID- 12654719 TI - The Gene Ontology Annotation (GOA) project: implementation of GO in SWISS-PROT, TrEMBL, and InterPro. AB - Gene Ontology Annotation (GOA) is a project run by the European Bioinformatics Institute (EBI) that aims to provide assignments of terms from the Gene Ontology (GO) resource to gene products in a number of its databases (http://www.ebi.ac.uk/GOA). In the first stage of this project, GO assignments have been applied to a data set representing the complete human proteome by a combination of electronic mappings and manual curation. This vocabulary has also been applied to the nonredundant proteome sets for all other completely sequenced organisms as well as to proteins from a wide range of organisms where the proteome is not yet complete. PMID- 12654720 TI - Chromosomal deletion formation system based on Tn5 double transposition: use for making minimal genomes and essential gene analysis. AB - In this communication, we describe the use of specialized transposons (Tn5 derivatives) to create deletions in the Escherichia coli K-12 chromosome. These transposons are essentially rearranged composite transposons that have been assembled to promote the use of the internal transposon ends, resulting in intramolecular transposition events. Two similar transposons were developed. The first deletion transposon was utilized to create a consecutive set of deletions in the E. coli chromosome. The deletion procedure has been repeated 20 serial times to reduce the genome an average of 200 kb (averaging 10 kb per deletion). The second deletion transposon contains a conditional origin of replication that allows deleted chromosomal DNA to be captured as a complementary plasmid. By plating cells on media that do not support plasmid replication, the deleted chromosomal material is lost and if it is essential, the cells do not survive. This methodology was used to analyze 15 chromosomal regions and more than 100 open reading frames (ORFs). This provides a robust technology for identifying essential and dispensable genes. PMID- 12654721 TI - Efficient high-throughput resequencing of genomic DNA. AB - Targeted resequencing of genomic DNA from organisms such as humans is an important tool enabling experimental access to variation within the species and between similar species. Taking full advantage of the reference genome sequences in designing robust, specific PCR assays and using stringent conditions, resequencing can be done efficiently without purification of the PCR product. By using a 10-fold greater amount of one primer when setting up the PCR initially in a new version of asymmetric PCR, one simply adds the rest of the sequencing reagents at the end of PCR and allows the sequencing reaction to proceed, with the excess PCR primer serving as the sequencing primer. We demonstrated that this streamlined protocol can be used with PCR products up to 1300 bp and had up to a 97% success rate in high-throughput analysis of allele frequencies for >30,000 single-nucleotide polymorphisms (SNPs). SNP primers and characterization results are provided at http://snp.wustl.edu. PMID- 12654722 TI - Computationally identifying novel NF-kappa B-regulated immune genes in the human genome. AB - Identifying novel NF-kappa B-regulated immune genes in the human genome is important to our understanding of immune mechanisms and immune diseases. We fit logistic regression models to the promoters of 62 known NF-kappa B-regulated immune genes, to find patterns of transcription factor binding in the promoters of genes with known immune function. Using these patterns, we scanned the promoters of additional genes to find matches to the patterns, selected those with NF-kappa B binding sites conserved in the mouse or fly, and then confirmed them as NF-kappa B-regulated immune genes based on expression data. Among 6440 previously identified promoters in the human genome, we found 28 predicted immune gene promoters, 19 of which regulate genes with known function, allowing us to calculate specificity of 93%-100% for the method. We calculated sensitivity of 42% when searching the 62 known immune gene promoters. We found nine novel NF kappa B-regulated immune genes which are consistent with available SAGE data. Our method of predicting gene function, based on characteristic patterns of transcription factor binding, evolutionary conservation, and expression studies, would be applicable to finding genes with other functions. PMID- 12654723 TI - LAGAN and Multi-LAGAN: efficient tools for large-scale multiple alignment of genomic DNA. AB - To compare entire genomes from different species, biologists increasingly need alignment methods that are efficient enough to handle long sequences, and accurate enough to correctly align the conserved biological features between distant species. We present LAGAN, a system for rapid global alignment of two homologous genomic sequences, and Multi-LAGAN, a system for multiple global alignment of genomic sequences. We tested our systems on a data set consisting of greater than 12 Mb of high-quality sequence from 12 vertebrate species. All the sequence was derived from the genomic region orthologous to an approximately 1.5 Mb region on human chromosome 7q31.3. We found that both LAGAN and Multi-LAGAN compare favorably with other leading alignment methods in correctly aligning protein-coding exons, especially between distant homologs such as human and chicken, or human and fugu. Multi-LAGAN produced the most accurate alignments, while requiring just 75 minutes on a personal computer to obtain the multiple alignment of all 12 sequences. Multi-LAGAN is a practical method for generating multiple alignments of long genomic sequences at any evolutionary distance. Our systems are publicly available at http://lagan.stanford.edu. PMID- 12654724 TI - A divergent INS protein in Caenorhabditis elegans structurally resembles human insulin and activates the human insulin receptor. AB - Caenorhabditis elegans contains a family of putative insulin-like genes proposed to regulate dauer arrest and senescence. These sequences often lack characteristic sequence features of human insulin essential for its folding, structure, and function. Here, we describe the structure and receptor-binding properties of INS-6, a single-chain polypeptide expressed in specific neurons. Despite multiple nonconservative changes in sequence, INS-6 recapitulates an insulin-like fold. Although lacking classical receptor-binding determinants, INS 6 binds to and activates the human insulin receptor. Its activity is greater than that of an analogous single-chain human insulin analog. PMID- 12654725 TI - The death domain kinase RIP has an essential role in DNA damage-induced NF-kappa B activation. AB - The transcription factor NF-kappaB is activated when cells are exposed to genotoxic stress. It has been suggested that DNA damage will trigger a cytoplasmic signaling that leads to the activation of IKK and NF-kappaB, but the signaling components upstream of IKK have not yet been identified. Here we report that the receptor interacting protein, RIP, is the IKK upstream component, essential for the activation of NF-kappaB by DNA damage. Also, our findings suggest that this NF-kappaB activation by DNA damage is not mediated by autocrine or TNF-R1 signaling pathway. In wild-type fibroblasts, DNA damage induced by agents such as adriamycin, campthothecin, and ionizing radiation induces NF kappaB activation. We found, however, that DNA damage failed to activate NF kappaB in RIP-/- fibroblasts. The induction of IkappaBalpha degradation by DNA damage was normal in TNF-R1-/-, TRAF2-/-, TRAF5-/- and FADD-/- fibroblasts or when de novo protein synthesis was blocked. More importantly, the reconstitution of RIP expression in RIP-/- cells restores DNA damage-induced NF-kappaB activation. We also found that RIP forms a complex with IKK in response to DNA damage. Therefore, our study provides a possible mechanism for the initiation of the cytoplasmic signaling to activate NF-kappaB in response to DNA damage. PMID- 12654726 TI - Essential role for caspase 8 in T-cell homeostasis and T-cell-mediated immunity. AB - Defects in death receptor-mediated apoptosis have been linked to cancer and autoimmune disease in humans. The in vivo role of caspase 8, a component of this pathway, has eluded analysis in postnatal tissues because of the lack of an appropriate animal model. Targeted disruption of caspase 8 is lethal in utero. We generated mice with a targeted caspase 8 mutation that is restricted to the T cell lineage. Despite normal thymocyte development in the absence of caspase 8, we observed a marked decrease in the number of peripheral T-cells and impaired T cell response ex vivo to activation stimuli. caspase 8 ablation protected thymocytes and activated T-cells from CD95 ligand but not anti-CD3-induced apoptosis, or apoptosis activated by agents that are known to act through the mitochondria. caspase 8 mutant mice were unable to mount an immune response to viral infection, indicating that caspase 8 deletion in T-cells leads to immunodeficiency. These findings identify an essential, cell-stage-specific role for caspase 8 in T-cell homeostasis and T-cell-mediated immunity. This is consistent with the recent identification of caspase 8 mutations in human immunodeficiency. PMID- 12654729 TI - Hospitalization, a risk factor for antibiotic-resistant Escherichia coli in the community? AB - OBJECTIVE: The impact of hospitalization on the prevalence of resistant Escherichia coli in the intestinal flora of patients admitted to the surgical wards of three Dutch university-affiliated hospitals was analysed prospectively. METHODS: Faecal samples were obtained on admission to the hospital, at the time of discharge, and 1 and 6 months after discharge. All samples were examined for resistance to nine antibiotic agents. RESULTS: For the total patient population, no significant differences in the prevalence of resistance were observed at the different sampling intervals, except for a significant decrease in cefazolin resistance between the time of discharge and 6 months after discharge (10% to 3%, P < 0.05). This decrease was mainly observed in patients from the university hospital Maastricht (azM), in which a significant decrease from 17% to 6% was detected (P < 0.05). Moreover, despite dissimilarities in patient characteristics and the marked variations in antibiotic use, no significant differences in the prevalence of antibiotic resistance were observed between the three hospitals, except for the overall higher prevalence of cefazolin-resistant E. coli in azM patients (P < 0.05). CONCLUSION: In this study, hospitalization did not seem to have any substantial effect on the prevalence of antibiotic-resistant E. coli at the different time intervals. However, as our study population consisted of surgical patients with a relatively moderate antibiotic use, and the prevalence of antibiotic resistance was only analysed for faecal E. coli, further investigation should be encouraged, as the understanding of the interaction between different resistance reservoirs is important for directing future intervention studies. PMID- 12654730 TI - Alginate-based oral drug delivery system for tuberculosis: pharmacokinetics and therapeutic effects. AB - Alginate microparticles were developed as oral sustained delivery carriers for antitubercular drugs in order to improve patient compliance. In the present study, pharmacokinetics and therapeutic effects of alginate microparticle encapsulated antitubercular drugs, i.e. isoniazid, rifampicin and pyrazinamide were examined in guinea pigs. Alginate microparticles containing antitubercular drugs were evaluated for in vitro and in vivo release profiles. These microparticles exhibited sustained release of isoniazid, rifampicin and pyrazinamide for 3-5 days in plasma and up to 9 days in organs. Peak plasma concentration (Cmax), Tmax, elimination half-life (t1/2e) and AUC0- infinity of alginate drugs were significantly higher than those of free drugs. The encapsulation of drug in alginate microparticles resulted in up to a nine-fold increase in relative bioavailability compared with free drugs. Chemotherapeutic efficacy of alginate drug microspheres against experimental tuberculosis showed no detectable cfu values at 1:100 and 1:1000 dilutions of spleen and lung homogenates. Histopathological studies further substantiated these observations, thus suggesting that application of alginate-encapsulated drugs could be useful in the effective treatment of tuberculosis. PMID- 12654728 TI - Translational control by TOR and TAP42 through dephosphorylation of eIF2alpha kinase GCN2. AB - Yeast protein kinase GCN2 stimulates the translation of transcriptional activator GCN4 by phosphorylating eIF2alpha in response to amino acid starvation. Kinase activation requires binding of uncharged tRNA to a histidyl tRNA synthetase related domain in GCN2. Phosphorylation of serine 577 (Ser 577) in GCN2 by another kinase in vivo inhibits GCN2 function in rich medium by reducing tRNA binding activity. We show that rapamycin stimulates eIF2alpha phosphorylation by GCN2, with attendant induction of GCN4 translation, while reducing Ser 577 phosphorylation in nonstarved cells. The alanine 577 (Ala 577) mutation in GCN2 (S577A) dampened the effects of rapamycin on eIF2alpha phosphorylation and GCN4 translation, suggesting that GCN2 activation by rapamycin involves Ser 577 dephosphorylation. Rapamycin regulates the phosphorylation of Ser 577 and eIF2alpha by inhibiting the TOR pathway. Rapamycin-induced dephosphorylation of Ser 577, eIF2alpha phosphorylation, and induction of GCN4 all involve TAP42, a regulator of type 2A-related protein phosphatases. Our results add a new dimension to the regulation of protein synthesis by TOR proteins and demonstrate cross-talk between two major pathways for nutrient control of gene expression in yeast. PMID- 12654731 TI - siRNAs: a new wave of RNA-based therapeutics. PMID- 12654732 TI - What have we learnt from community-acquired infections in Hong Kong? AB - This study was initiated throughout Hong Kong, to reveal the characteristics of community-acquired infections. All specimens collected by general practitioners from infected patients were followed prospectively, and those that were culture positive were analysed. Four thousand seven hundred and forty-one specimens were collected from 3977 patients by 89 doctors from July 2000 to October 2001. The most common specimens were throat swabs (33%), urine (26%) and sputa (16%). The average culture-positive rate was 28%. The most common organisms were Escherichia coli (18%), beta-haemolytic streptococci (15%) and Staphylococcus aureus (12%). Fluoroquinolone resistance was relatively high (up to 35%) in organisms commonly causing urinary tract infection (E. coli, Proteus and Morganella). Although none of the pneumococci was resistant to penicillin 1 mg/L, the proportion with intermediate resistance (0.1-1 mg/L) was alarming (81%). There were three strains of methicillin-resistant S. aureus. A decrease in ampicillin resistance but a high prevalence of macrolide resistance were noted in Haemophilus influenzae. All Neisseria gonorrhoeae isolates were resistant to penicillin, up to 79% to the fluoroquinolones, 15% to spectinomycin, but all were susceptible to ceftriaxone. Respiratory pathogens (Streptococcus pneumoniae, beta-haemolytic streptococci and H. influenzae) were relatively susceptible to the newer fluoroquinolones (0-2%, 0.5-6% and 2% resistant, respectively) or third-generation cephalosporins (0-2% resistant). The distribution of organisms and their antibiotic resistance varied over time. Thus frequent surveillance is needed to provide information on the drugs of choice for different infections. PMID- 12654727 TI - daf-28 encodes a C. elegans insulin superfamily member that is regulated by environmental cues and acts in the DAF-2 signaling pathway. AB - In Caenorhabditis elegans, the decision to enter a developmentally arrested dauer larval stage is triggered by a combination of signals from sensory neurons in response to environmental cues, which include a dauer pheromone. These sensory inputs are coupled to the parallel DAF-2/insulin receptor-like and DAF-7/TGFbeta like signaling pathways. Although sensory inputs have been shown to physiologically regulate DAF-7/TGFbeta expression, no such regulation of insulin like ligands in the DAF-2 pathway has been reported. We show here that daf-28 encodes an insulin-like protein, which when mutated causes dauer arrest and down regulation of DAF-2/IR signaling. A daf-28GFP fusion gene is expressed in ASI and ASJ, two sensory neurons that regulate dauer arrest. daf-28GFP expression in ASI and ASJ is down-regulated under dauer-inducing conditions and in mutants of DAF 11/guanylyl cyclase, a predicted component of the dauer-pheromone-sensing pathway. Thus, daf-28 expression in sensory neurons is regulated by the environmental cues that normally trigger dauer arrest. Among the 38 C. elegans insulin genes, daf-28 is so far the only insulin mutant to affect dauer arrest. daf-28 was revealed from this functional redundancy by a dominant-negative allele that disrupts a probable proteolytic processing site required for insulin maturation. This DAF-28 mutant is likely to be poisonous to wild-type DAF-28 and other insulins. PMID- 12654733 TI - Mutations in gyrA and parC genes in nalidixic acid-resistant Escherichia coli strains from food products, humans and animals. AB - Mutations in quinolone targets were analysed in 80 unrelated nalidixic acid resistant (NALR) Escherichia coli strains whose nalidixic acid and ciprofloxacin MICs ranged from 32 to >256 mg/L and 0.03-64 mg/L, respectively. These strains were isolated from food products (23) and faecal samples from humans (15) and healthy animals (42). Thirteen nalidixic acid-susceptible (NALS) E. coli strains were also analysed. Mutations in gyrA and parC genes were studied by PCR and sequencing. No amino acid changes were detected in GyrA or ParC proteins of the 13 NALS strains. A single change in the GyrA protein was detected in all 61 NALR strains with ciprofloxacin MICs Leu (54), Ser-83-->Ala (one), Ser-83-->Val (one), Asp-87-->Asn (two), Asp-87-->Tyr (two) and Asp-87-->Gly (one). A double change in GyrA was found in 18 of 19 NALR strains with ciprofloxacin MICs >/= 4 mg/L. Amino acid substitutions were Ser-83-->Leu, with an additional change [Asp-87-->Asn (15), Asp-87-->Tyr (two) or Asp-87-->His (one)]. The remaining strain (ciprofloxacin MIC 4 mg/L) showed a single Ser-83-->Leu substitution. In respect of the ParC protein, a single change at Ser-80 or Glu-84 was found in 25 of 42 strains, with ciprofloxacin MICs ranging from 0.5 to 32 mg/L. A double substitution (Ser-80-->Ile and Glu-84-->Gly) was found in one strain (ciprofloxacin MIC 64 mg/L). No amino acid changes were detected in the GyrB protein of 18 NALR strains. PMID- 12654734 TI - An integron cassette encoding erythromycin esterase, ere(A), from Providencia stuartii. AB - We have mapped the variable region of the two class 1 integrons found in the multiresistant strain Providencia stuartii 1723. Integron 1 contains a new arrangement of gene cassettes, aacA4-aadB-aadA1, conferring resistance to all aminoglycosides used for clinical treatment. Integron 2 contains a variant of the gene cassette ere(A), coding for an erythromycin esterase, whose nucleotide sequence shares 93.7% DNA identity with ere(A) from Escherichia coli BM2195 plasmid pIP1100. PMID- 12654735 TI - Structure-activity relationships for a series of peptidomimetic antimicrobial prodrugs containing glutamine analogues. AB - Synthetic glutamine analogues such as N3-(4-methoxyfumaroyl)-l-2,3 diaminopropanoic acid (FMDP) inhibit purified glucosamine-6-phosphate synthase, an intracellular enzyme that is essential for microbial cell wall synthesis, but they are inactive against intact organisms because they cannot enter the cell. However, when the analogues are linked to a peptide they can be actively transported, and FMDP peptidomimetics show broad-spectrum antimicrobial activity. To characterize this process in more detail, the antibacterial activities of various synthetic peptidomimetics containing glutamine analogues have been determined against isogenic strains of Escherichia coli in which one or more of its three peptide transporters Dpp, Opp and Tpp have been mutated. In addition, their affinities for DppA and OppA, the binding-protein components of the transporters, have been measured. In general, antibacterial activities against the various transport mutants correlated with binding to DppA and OppA. Xaa-FMDP compounds have greater activities than FMDP-Xaa analogues. To explore structure activity relationships for the peptidomimetics, molecular modelling was used to determine the conformational forms they adopt in solution. The relative bioactivities of the peptidomimetics correlated with the percentage of conformers that had backbone torsions matching those previously defined for the molecular recognition templates of the peptide transporters. However, the large size of the N-terminal residue in the FMDP-Xaa analogues appears to interfere with transport and thus to limit antibacterial activity. Overall, the results provide the structural rationale for the identification in silico of analogues with optimal bioactivities, which decreases the need for extensive chemical syntheses and testing. PMID- 12654736 TI - Antibiotic susceptibility and serotypes of Streptococcus pneumoniae isolates from Hungary. AB - OBJECTIVE: Hungary has reported one of the highest incidences of penicillin resistance in Streptococcus pneumoniae in Europe since the 1970s and is still cited accordingly. However, since the end of the 1990s the resistance of pneumococci in Hungary has not been investigated. In this study we assessed the current situation, particularly to establish whether the incidence of resistance is increasing and if this could be related to the spread of specific strain types. METHODS: Isolates of S. pneumoniae (n = 304) were collected by five diagnostic laboratories in Hungary in 2000-2002. Their identity was confirmed and their susceptibilities to 16 antibiotics were determined by the agar dilution method according to NCCLS guidelines. Representative strains were serotyped (n = 112). RESULTS AND CONCLUSIONS: We found significantly lower resistance rates for penicillin compared with the data previously reported from Hungary, but the intermediate resistance was high, at 37%. Macrolide resistance was a bigger problem ( approximately 40% for erythromycin), although there was full susceptibility to telithromycin. The strains with the highest MICs were isolated from carriers and young children. The fluoroquinolones were very effective, especially moxifloxacin and gatifloxacin. There was full susceptibility to vancomycin and linezolid. We found inconsistencies with previous reports in the survey of the resistance and identification of S. pneumoniae in the country. The serotype distribution of the isolates showed a much greater diversity than had previously been reported; however, there was correlation between serotype and resistance. PMID- 12654737 TI - Therapeutic potential of neutralizing antibodies in the treatment of HIV-1 infection. PMID- 12654738 TI - Induction of mexCD-oprJ operon for a multidrug efflux pump by disinfectants in wild-type Pseudomonas aeruginosa PAO1. AB - Induction of the MexCD-OprJ multidrug efflux pump was investigated in wild-type Pseudomonas aeruginosa PAO1. MexCD-OprJ was induced by clinically important disinfectants such as benzalkonium chloride and chlorhexidine gluconate, and by some cytotoxic agents such as tetraphenylphosphonium chloride, ethidium bromide and rhodamine 6G. MexCD-OprJ was not induced by norfloxacin, tetracycline, chloramphenicol, streptomycin, erythromycin or carbenicillin, although they are substrates for the pump. Cells of PAO1 showed increased resistance to norfloxacin when grown in the presence of the inducers of the mexCD-oprJ operon mentioned above. These results indicate that MexCD-OprJ plays an important role in intrinsic multidrug resistance in wild-type P. aeruginosa in hospitals where disinfectants are used frequently. PMID- 12654739 TI - Inhibition of human immunodeficiency virus 1 replication in vitro by a self stabilized oligonucleotide with 2'-O-methyl-guanosine-uridine quadruplex motifs. AB - OBJECTIVES: Given that the guanosine-quadruplex may have a role in blocking the interaction between gp120 and CD4, we describe here the design of a highly nuclease-resistant dimeric hairpin guanosine-quadruplex, [Gm3Um4Gm3-s], containing the 2'-O-methyl groups on the nucleoside and sulphur groups on the internucleotidic bonds, and its anti-HIV-1 activity in cultured cells. METHODS: The unmodified and modified oligonucleotides were chemically synthesized. The anti-HIV activities of test compounds on HIV-1 infection were determined by protection against HIV-1-induced cytopathic effects. The mechanism of action of the oligonucleotides was determined by virus binding and detection [anti-CD4 monoclonal antibody (MAb) and anti-V3 MAb] assays. RESULTS: Gm3Um4Gm3-s was highly nuclease resistant, had significantly higher anti-HIV-1 activity than dG3T4G3-s, dG10-s and Gm10-s, and blocked the interaction between gp120 and CD4. CONCLUSION: The anti-HIV-1 activity of this oligonucleotide was increased when the phosphodiester and 2'-hydroxyl groups on the oligonucleotide backbones were replaced with a phosphorothioate and 2'-O-methyl backbone; thus Gm3Um4Gm3-s may inhibit HIV-1 infection, at least in part, by blocking the interaction between gp120 and CD4. PMID- 12654740 TI - Amelioration of nephropathy in mice expressing HIV-1 genes by the cyclin dependent kinase inhibitor flavopiridol. AB - Cumulative evidence suggests that human immunodeficiency virus-associated nephropathy (HIVAN), the third leading cause of end-stage renal disease in African-Americans, may respond to therapeutic strategies that interrupt HIV-1 expression in infected renal epithelium. We recently demonstrated that suppression of HIV-1 transcription in infected glomerular visceral epithelial cells by flavopiridol, a small-molecule inhibitor of the cyclin-dependent kinases required for HIV-1 promoter activity, reversed HIV-induced proliferation and dedifferentiation in vitro. To address whether flavopiridol could ameliorate HIV induced renal disease, we utilized a well-established HIV-1 NL4-3 transgenic mouse model of HIVAN. HIV-1 proviral transgene expression in whole kidney was markedly suppressed by a 20 day treatment with flavopiridol. Following treatment, histopathological, serological and urinary indices of nephrosis were normalized in flavopiridol-treated but not in vehicle-treated transgenics. Microarray analysis showed that 82% of the dysregulated genes in HIVAN kidney were normalized to control levels by flavopiridol, whereas continued dysregulation of most of the remaining 18% was attributable to an effect from flavopiridol alone. These results demonstrate for the first time that targeting the cyclin-dependent kinases that support HIV-1 expression can ameliorate HIV-induced disease in an animal model. PMID- 12654741 TI - A high performance liquid chromatography (HPLC) assay for linezolid in continuous ambulatory peritoneal dialysis fluid (CAPDF). PMID- 12654742 TI - The role of genomics in antimicrobial discovery. PMID- 12654743 TI - Stability and in vitro efficacy of antibiotic-heparin lock solutions potentially useful for treatment of central venous catheter-related sepsis. AB - OBJECTIVES: Increasing numbers of patients for whom infection is a major risk are dependent on central venous catheters. Antibiotic-anticoagulant locks may have a role in preventing or treating catheter-related infections. The aim of this study was to determine the in vitro stability and efficacy of antibiotic-heparin lock solutions. METHODS: Candidate antibiotics (amikacin, ciprofloxacin, flucloxacillin, gentamicin, linezolid, teicoplanin) were investigated in vitro, either individually or in combination, in solution with heparin. The solutions were initially tested for visual precipitation. The efficacy of stable solutions and taurolidine was then tested in a catheter model bioassay system against microorganisms commonly encountered in catheter-related septicaemia. RESULTS: In general, lower concentrations of heparin (/= 10 mg/L) during the treatment period substantially increased from 3.2% on day 2, to 35%, 70%, 90% and approximately 95% on days 4, 7, 11 and 15, respectively. The findings suggest that optimal teicoplanin therapy was achieved only after at least 4, and probably 7, days of therapy in most cases, mainly because of a failure to use an appropriate loading dose. Among the possible causes for the reluctance to use a loading dose, concern over the potential nephrotoxicity of teicoplanin was a major factor. We conclude that loading doses of teicoplanin (6 mg/kg every 12 h for at least three doses) must be considered mandatory in all patients, regardless of their renal function, to enable optimal drug concentrations to be achieved early in the treatment period. Subsequently, TDM is important to ensure that dose regimens are optimized to the individual requirements of the patients. PMID- 12654758 TI - Role of the acetyltransferase AAC(6')-Iz modifying enzyme in aminoglycoside resistance in Stenotrophomonas maltophilia. AB - Stenotrophomonas maltophilia is an emerging nosocomial pathogen that displays high-level intrinsic resistance to multiple antibiotics including aminoglycosides. A gene [aac(6')-Iz] encoding an aminoglycoside-modifying enzyme, AAC(6')-Iz acetyltransferase, was recently cloned and sequenced in S. maltophilia, but its importance with respect to aminoglycoside resistance in this organism was not determined. Using a homologous gene replacement approach, mutants carrying unmarked chromosomal deletions of the aac(6')-Iz gene were constructed in wild-type and in vitro-selected aminoglycoside-resistant S. maltophilia. AAC(6')-Iz-deficient mutants derived from both wild-type and aminoglycoside-resistant strains displayed an increase in susceptibility to amikacin, netilmicin, sisomicin and tobramycin (4- to 32-fold decrease in MICs), known substrates for AAC(6')-I enzymes. The cloned aac(6')-Iz gene restored the aminoglycoside resistance of the aac(6')-Iz mutants, and could also confer aminoglycoside resistance upon Escherichia coli. To assess the significance of the aac(6')-Iz gene with respect to the aminoglycoside resistance of clinical strains, its distribution was assessed in 65 clinical isolates from two hospitals. Using PCR, Southern hybridization, RT-PCR and/or nucleotide sequencing, the aac(6')-Iz gene was identified in 57% of the isolates. Susceptibility tests indicated a good correlation between the presence of the aac(6')-Iz gene and the resistance to tobramycin, netilmicin and sisomicin in these strains. These results indicate that the aac(6')-Iz gene is an important contributor to aminoglycoside resistance in clinical strains of S. maltophilia, particularly to tobramycin. PMID- 12654759 TI - In vitro effect on Cryptosporidium parvum of short-term exposure to cathelicidin peptides. AB - Two laboratory methods, a cell culture system and double fluorogenic staining, were used to study the viability and infective ability of Cryptosporidium parvum sporozoites and oocysts after short-term exposure to four cathelicidin peptides. The compounds, SMAP-29, BMAP-28, PG-1 and Bac7(1-35), exerted a strong cytotoxic effect on sporozoites, but did not affect the viability and function of oocysts consistently. Overall, in the sporozoite series, a percentage of the viable population decreased rapidly to less than detectable levels after 15 and 60 min exposure to the peptides at concentrations of 100 and 10 micro g/mL, respectively. In the oocyst series, no compound produced complete inhibition of parasite growth: 60-85% of the oocyst population was viable after 180 min exposure at 100 micro g/mL. SMAP-29 exerted the highest activity against both sporozoites and oocysts. PMID- 12654760 TI - Isolation, excystation and axenization of Giardia lamblia isolates: in vitro susceptibility to metronidazole and albendazole. AB - From 53 samples of human faeces containing Giardia lamblia cysts, 18 isolates were successfully excysted in vitro, and cultivated axenically in TYI-S-33 modified medium. The in vitro effects of metronidazole and albendazole on these isolates were evaluated by the trophozoite adherence inhibition method. The IC50 was between 2.4 and 11.5 micro M for metronidazole and 0.027 and 0.192 micro M for albendazole. These IC50 values were similar to those found for the ATCC 30888 and 30957 reference isolates. All isolates were susceptible to the antiparasitic drugs tested. These results suggest that resistance of G. lamblia to metronidazole and albendazole does not seem to be a significant problem in our population. PMID- 12654762 TI - AUC(0-t)/MIC is a continuous index of fluoroquinolone exposure and predictive of antibacterial response for Streptococcus pneumoniae in an in vitro infection model. AB - OBJECTIVE: To conduct a comprehensive pharmacodynamic analysis of moxifloxacin and levofloxacin against Streptococcus pneumoniae in an in vitro infection model. METHODS: In dose escalation studies, single doses with peak concentrations equivalent to 1 x, 2 x, 4 x, 8 x, 16 x and 32 x MIC against two isolates of S. pneumoniae were studied over 24 h. Traditional pharmacodynamic indices, including peak concentration divided by MIC (peak/MIC), time of concentration above MIC (T > MIC) and AUC24/MIC, were estimated for all regimens. As a continuous index of fluoroquinolone exposure, AUC0-t/MIC was also calculated, as AUC from time 0 to 1, 2 and 6 h divided by MIC. Correlations between pharmacodynamic indices and antibacterial effects were examined using linear and non-linear methods. In validation experiments, the pharmacodynamic model was used to predict bacterial kill curves, produced by simulated clinical doses of moxifloxacin and levofloxacin against two other S. pneumoniae isolates. RESULTS: Peak/MIC was most predictive of early bacterial kill, whereas T > MIC was significantly associated with final bacterial counts at 24 h. Antibacterial effects were bacteriostatic when T > MIC was 48% and bactericidal when values exceeded 55%. AUC0-t/MIC was strongly associated with bacterial kill throughout the dosing interval. Bactericidal activity and bacterial eradication were associated with AUC0-t/MICs of 28 and 135, respectively. AUC0-t/MIC was also highly predictive of bacterial kill curves produced by simulated clinical doses of moxifloxacin and levofloxacin (precision 0.36 log10 cfu/mL, bias 0.02 log10 cfu/mL). CONCLUSION: This study demonstrated the novel application of AUC0-t/MIC as a continuous index of antibiotic activity, and provided extensive characterization of fluoroquinolone pharmacodynamics against S. pneumoniae. PMID- 12654761 TI - Effects of the antimitotic natural product dolastatin 10, and related peptides, on the human malarial parasite Plasmodium falciparum. AB - Microtubule inhibitors from several chemical classes can block the growth and development of malarial parasites, reflecting the importance of microtubules in various essential parasite functions. With the spread of antimalarial drug resistance, there is an urgent need for new approaches to the chemotherapy of this devastating disease. We investigated the effects of two naturally occurring marine peptides, dolastatin 10 and dolastatin 15, and 10 synthetic dolastatin 10 based compounds (auristatins), on cultured malarial parasites of the species most lethal to humans, Plasmodium falciparum. Dolastatin 10 was a more potent inhibitor of P. falciparum than any other previously described microtubule inhibitor, with a median inhibitory concentration (IC50) of 10-10 M. Dolastatin 15 was less active, and compounds of the auristatin series had various potencies. Comparison of the concentrations required to inhibit P. falciparum and mammalian cell proliferation showed that the orders of potency were not the same. Dolastatin 10 and auristatin PE caused arrested nuclear division and apparent disassembly of mitotic microtubular structures in the parasite. The effects of these agents were, superficially at least, similar to those of vinblastine but different from those of paclitaxel. These studies indicate that compounds binding in the 'Vinca domain' of tubulin can be highly potent antimalarial agents. PMID- 12654763 TI - Community-acquired respiratory tract infections caused by resistant pneumococci: clinical and bacteriological efficacy of the ketolide telithromycin. AB - The incidence of community-acquired respiratory tract infections caused by Streptococcus pneumoniae exhibiting antibacterial resistance has increased dramatically in recent years. Telithromycin is the first of a new class of antibacterials, the ketolides, which have been developed specifically to provide effective treatment for these infections. Data were analysed from 3935 patients who had participated in one Japanese Phase II study and 11 US/global Phase III studies in three indications: community-acquired pneumonia, acute exacerbations of chronic bronchitis or acute sinusitis. Patients received either telithromycin 800 mg once daily or a comparator antibacterial. S. pneumoniae isolates considered to be causative for infection were tested for susceptibility to penicillin G and erythromycin A. In per-protocol analyses, telithromycin showed a high level of clinical efficacy against S. pneumoniae, with clinical cure rates of 92.8% for all isolates, 91.7% for those with reduced susceptibility to penicillin G and 86.0% for those with reduced susceptibility to erythromycin A. Bacterial eradication rates were consistent with the clinical outcomes. High rates of clinical cure and bacterial eradication were also observed for infections caused by isolates demonstrating high-level resistance to erythro mycin A [MICs >/= 512 mg/L: 100% (13/13) clinical cure, 100% (13/13) bacterial eradication]. These results support the use of telithromycin as a first-line oral therapy for the treatment of community-acquired respiratory tract infections caused by S. pneumoniae with reduced susceptibility to penicillin G and erythromycin A. PMID- 12654764 TI - Microbiological efficacy of ABT-773 (cethromycin) for the treatment of community acquired pneumonia due to Chlamydia pneumoniae. AB - Nasopharyngeal specimens for culture of Chlamydia pneumoniae were obtained from patients with community-acquired pneumonia enrolled in a randomized study comparing the novel ketolide antibiotic ABT-773 at a dose of 150 mg once a day to 150 mg twice a day, by mouth for 10 days. C. pneumoniae was eradicated from the nasopharynx of 10 of 10 (100%) microbiologically evaluable patients. MICs and MBCs for 13 isolates of C. pneumoniae from 12 patients obtained before and after therapy were performed against ABT-773. The MIC90 and MBC90 of ABT-773 were 0.015 mg/L. PMID- 12654766 TI - Interaction of plasmid and host quinolone resistance. PMID- 12654765 TI - Natural antimicrobial susceptibilities of strains of 'unusual' Serratia species: S. ficaria, S. fonticola, S. odorifera, S. plymuthica and S. rubidaea. AB - The natural susceptibility to 71 antibiotics of 104 Serratia strains of Serratia ficaria (n = 15), Serratia fonticola (n = 18), Serratia odorifera (n = 16), Serratia plymuthica (n = 32) and Serratia rubidaea (n = 23) was examined. MICs were determined using a microdilution procedure in IsoSensitest broth for all the strains and in cation-adjusted Mueller-Hinton broth for some strains. With few exceptions, all species tested were uniformly naturally resistant to penicillin G, oxacillin, cefazolin, cefuroxime, all tested macrolides, lincosamides, streptogramins, glycopeptides, fusidic acid and rifampicin, and naturally sensitive to several aminoglycosides, piperacillin, piperacillin/tazobactam, carbapenems, some cephalosporins, fluoroquinolones and folate-pathway inhibitors. Major species-related differences in natural susceptibility affecting clinical assessment criteria were seen with tetracyclines, some aminoglycosides, aminopenicillins, ticarcillin, cefaclor, loracarbef, cefoxitin, pipemidic acid, chloramphenicol, nitrofurantoin and fosfomycin. Differences in susceptibility dependent on the medium were seen with macrolides, tetracycline, fosfomycin and some beta-lactams. The natural antibiotic susceptibility patterns suggest novel species-specific mechanisms of antibiotic resistance. Uncharacterized species specific aminoglycoside-modifying enzymes and multidrug efflux systems affecting tetracyclines, quinolones and chloramphenicol are probably responsible for some of the phenotypes observed. The natural amoxicillin sensitivity of several strains of some species combined with natural resistance to some narrow-spectrum cephalosporins indicate the expression of naturally occurring beta-lactamases with unique substrate profiles. beta-Lactamases of representative strains of each species were characterized phenotypically and genotypically. It was shown that all species expressed naturally occurring AmpC beta-lactamases and, with respect to S. fonticola, also a species-specific class A beta-lactamase. Inducibility of these enzymes was shown in all species with the exception of S. rubidaea and four of five strains of S. plymuthica. PMID- 12654767 TI - Clinical features and in vitro antimicrobial susceptibilities of community acquired Klebsiella pneumoniae meningitis in Taiwan. AB - Twenty-seven adult patients were identified as having community-acquired Klebsiella pneumoniae meningitis. The K. pneumoniae isolates, collected from cerebrospinal fluid samples, were tested for in vitro antimicrobial susceptibilities. The prognostic factors of these 27 patients were also analysed. All of the third- and fourth-generation cephalosporins tested, as well as monobactam, carbapenem and ciprofloxacin, had good activities against the isolated K. pneumoniae strains. None of the clinical isolates was detected as being an extended-spectrum beta-lactamase-producing pathogen. Among the third- and fourth-generation cephalosporins, ceftizoxime, cefepime, ceftriaxone and cefotaxime had superior activities, with MIC90s about four- to eight-fold lower than those of ceftazidime and moxalactam. Mortality rates of patients classified by different antimicrobial agents were as follows: ceftazidime 38% (8/21) and cefepime 16.7% (1/6). The presence of septic shock and the initial level of consciousness at the start of appropriate antimicrobial therapy were the major determinants of survival and neurological outcomes in these 27 patients. Early diagnosis and choice of appropriate antibiotics according to antimicrobial susceptibilities may improve therapeutic outcomes. PMID- 12654768 TI - Nalidixic acid-resistant strains of Salmonella showing decreased susceptibility to fluoroquinolones in the mid-west region of the Republic of Ireland. PMID- 12654769 TI - In vitro activity of linezolid alone and in combination with gentamicin, vancomycin or rifampicin against methicillin-resistant Staphylococcus aureus by time-kill curve methods. AB - The in vitro activity of the oxazolidinone linezolid was studied alone and in combination with three antibiotics acting on different cellular targets. Oxazolidinones are bacterial protein synthesis inhibitors that act at a very early stage by preventing the formation of the initiation complex. Combinations of linezolid with gentamicin, vancomycin or rifampicin were evaluated against four methicillin-resistant Staphylococcus aureus strains, using killing curves in conjunction with scanning electron microscopy. Time-kill curves were performed over 24 h using an inoculum of 5 x 10(6)- 1 x 10(7) cfu/mL. Linezolid was studied at concentrations of 1 x, 4 x and 8 x MIC, with partner drugs at 8 x MIC. Addition of linezolid resulted in a decrease of antibacterial activity for gentamicin and vancomycin, and linezolid was antagonistic to the early bactericidal activity of gentamicin. Linezolid, in combination with rifampicin, showed an additive interaction for susceptible strains and inhibited rifampicin resistant variants. Linezolid plus rifampicin appeared to be the most active combination against methicillin-resistant S. aureus strains in time-kill experiments. PMID- 12654770 TI - In vitro susceptibility to a new antimalarial organometallic analogue, ferroquine, of Plasmodium falciparum isolates from the Haut-Ogooue region of Gabon. AB - OBJECTIVES: To assess the activity of a new organometallic chloroquine analogue, ferroquine, against numerous Plasmodium falciparum isolates from Gabon. METHODS: The in vitro susceptibility of 116 P. falciparum isolates to chloroquine and ferroquine was assessed using the isotopic microtest. All isolates were from outpatients in the Franceville and Bakoumba medical centres in the province of Haut-Ogooue, south-east Gabon. RESULTS: The in vitro resistance to chloroquine was 51.8% in Franceville and 96.7% in Bakoumba. The IC50 geometric mean (95% CI) of ferroquine against isolates in Franceville was 16.0 (14.4-17.8) nM, with individual values ranging from 1.0 to 47.0 nM; in Bakoumba it was 27.9 (23.4 33.2) nM, with individual values ranging from 1.0 to 62.0 nM. Compared with chloroquine, ferroquine was 5.3 times more active on isolates susceptible to chloroquine, and 13.3 times more active on isolates resistant to chloroquine. A weak positive correlation was observed between responses of these two drugs, but too low to demonstrate cross-resistance. CONCLUSIONS: Ferroquine may be useful as an alternative drug for treating chloroquine-resistant malaria. PMID- 12654771 TI - In vitro development of resistance to a novel fluoroquinolone, DW286, in methicillin-resistant Staphylococcus aureus clinical isolates. AB - In vitro development of resistance to a novel fluoroquinolone, DW286, as well as to ciprofloxacin, gemifloxacin, sparfloxacin and trovafloxacin, was investigated in eight methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates. The strains were subcultured in subinhibitory concentrations of each agent during a 50 day period. Subculturing in most agents led to the selection of 37 mutants with increased MICs. The DW286 MICs were increased from 0.004-0.031 to 0.125-0.5 mg/L in five strains after 13-47 passages, and were not increased in three strains. The ciprofloxacin, gemifloxacin, sparfloxacin and trovafloxacin-selected mutants showed relatively weak cross-resistance to DW286. DNA sequencing analyses of all of the selected mutants revealed a few point mutations responsible for the high level of resistance, but actually these variations did not confer high resistance to fluoroquinolones. In the presence of reserpine, an inhibitor of the Gram-positive efflux pump, of 36 mutants 22 had two- to 16-fold lower ciprofloxacin MICs, and 20 had two- to 16-fold lower gemifloxacin MICs. However, sparfloxacin, trovafloxacin and DW286 were not good substrates for efflux pumps. PMID- 12654772 TI - On the use of DNA vaccines for the prophylaxis of mycobacterial diseases. PMID- 12654773 TI - Flagella are virulence determinants of Burkholderia pseudomallei. AB - Burkholderia pseudomallei, a facultatively intracellular pathogen, is a flagellated and motile gram-negative bacterium and is the causative agent of melioidosis in humans. Flagella are commonly recognized as important virulence determinants expressed by bacterial pathogens since the motility phenotype imparted by these organelles often correlates with the ability of an organism to cause disease. We used a virulent isolate of B. pseudomallei, KHW, to construct an isogenic deletion mutant with a mutation in the flagellin gene (fliC) by gene replacement transposon mutagenesis. The KHWDeltafliCKm mutant was aflagellate and nonmotile in semisolid agar. The isogenic KHWDeltafliCKm mutant was not impaired in terms of the ability to invade and replicate in cultured human lung cells compared with the wild type. It was also equally virulent in slow-killing assays involving Caenorhabditis elegans, but it was avirulent during intranasal infection of BALB/c mice. Very few bacteria, if any, were isolated from the lungs and spleens of KHWDeltafliCKm-infected mice. In contrast, the bacterial loads in the lungs and spleens were similar in mice infected with KHW and in mice infected with the complemented mutant, KHWDeltafliCKm/pUCP28TfliC. Unlike the Syrian hamster or diabetic rat models of infection, the B. pseudomallei flagellin was also a virulence factor during intraperitoneal infection of BALB/c mice. In this study, all animals infected with KHWDeltafliCKm remained healthy and did not succumb to disease regardless of the route of infection. The flagellum is therefore an important and necessary virulence determinant of B. pseudomallei during intranasal and intraperitoneal infection of mice. PMID- 12654774 TI - Interleukin-18 impairs the pulmonary host response to Pseudomonas aeruginosa. AB - Interleukin-18 (IL-18) is a potent cytokine with many different proinflammatory activities. To study the role of IL-18 in the pathogenesis of Pseudomonas pneumonia, IL-18-deficient (IL-18(-/-)) and wild-type mice were intranasally inoculated with Pseudomonas aeruginosa. IL-18 deficiency was associated with reduced outgrowth of Pseudomonas in the lungs and diminished dissemination of the infection. In addition, pulmonary inflammation (histopathology) and levels of tumor necrosis factor alpha, IL-6, and macrophage inflammatory protein-2 in lungs and plasma were lower in IL-18(-/-) mice. Consistent with results obtained for IL 18(-/-) mice, treatment of wild-type mice with a neutralizing IL-18 binding protein-immunoglobulin G Fc fusion construct also attenuated outgrowth of Pseudomonas compared with that for mice treated with a control protein. These results demonstrate that the presence of endogenous IL-18 activity facilitates inflammatory responses in the lung during Pseudomonas pneumonia, concurrently impairing bacterial clearance. PMID- 12654775 TI - Prevalence and distribution of adhesins in invasive non-type b encapsulated Haemophilus influenzae. AB - Adhesion to the respiratory epithelium plays an important role in Haemophilus influenzae infection. The distribution of H. influenzae adhesins in type b and nontypeable strains has been characterized, but little is known about the prevalence of these factors in non-type b encapsulated strains. We analyzed 53 invasive type a, type e, and type f strains for the presence of hap, hia, hmw, and hif genes; Hap, Hia, and HMW1/2 adhesins; and hemagglutinating pili. The hap gene was ubiquitous, and homologs of hmw and hia were present in 7 of 53 (13.2%) and 45 of 53 (84.9%) strains, respectively. Hap was detected in 28 of 45 (62.2%) hap(+) strains, HMW1/2 was detected in 5 of 7 (71.4%) hmw(+) strains, and Hia was detected in 31 of 45 (68.8%) hia(+) strains. The hif gene cluster was present in 26 of 53 strains (49.1%), and 21 of 26 hif(+) strains (80.8%) agglutinated (HA) red blood cells. Nine isolates exhibited HA but lacked the hif gene cluster. The distribution of adhesin genes correlated with the genetic relatedness of the strains. Strains belonging to one type a clonotype and the major type e clonotype possessed hia but lacked the hif cluster. Strains belonging to the second type a clonotype possessed both hia and hif genes. All type f strains belonging to the major type f clonotype possessed hia and lacked hifB. Although the specific complement of adhesin genes in non-type b encapsulated H. influenzae varies, most invasive strains express Hap and Hia, suggesting these adhesins may be especially important to the virulence of these organisms. PMID- 12654776 TI - Effect of interleukin-8 and granulocyte colony-stimulating factor on priming and activation of bovine neutrophils. AB - Neutrophils are important effector cells in innate and acquired immunity, but the magnitude and character of their phagocytic and bactericidal responses depend on cues derived from mediators in the local microenvironment. This study investigated the effect of bovine interleukin-8 (IL-8) and granulocyte colony stimulating factor (G-CSF) on priming and activation of bovine neutrophils in vitro and in vivo. Neutrophils were isolated from blood and cultured for up to 18 h, with or without cytokines, and then Mannheimia haemolytica-induced oxidative burst and phagocytosis of Staphylococcus aureus were measured by flow cytometry. Neither IL-8 nor G-CSF directly triggered an oxidative burst, but incubation with these cytokines for 18 h primed neutrophils for a greater oxidative burst triggered by M. haemolytica and for enhanced uptake of S. aureus. The maximal response was observed when neutrophils were incubated with both cytokines together, at concentrations of 200 ng/ml for G-CSF and 400 ng/ml for IL-8. The IL 8-induced priming effect was reduced by treatment with a neutralizing antibody to IL-8, and was not attributed to endotoxin contamination. Instillation of IL-8 into the lung using a bronchoscope induced neutrophil recruitment within 18 h. Neutrophils from IL-8-treated lung showed dose-dependent enhancement of the oxidative burst triggered by M. haemolytica. Histologically, neutrophils filled alveoli and bronchioles, and scattered macrophages contained neutrophils with morphological features of apoptosis. Thus, prolonged in vitro or in vivo exposure to IL-8 and/or G-CSF enhances the subsequent oxidative burst and phagocytic responses of bovine neutrophils. PMID- 12654777 TI - Cross-reactivity of antibodies against PorA after vaccination with a meningococcal B outer membrane vesicle vaccine. AB - The cross-reactivity of PorA-specific antibodies induced by a monovalent P1.7-2,4 (MonoMen) and/or a hexavalent (HexaMen) meningococcal B outer membrane vesicle vaccine (OMV) in toddlers and school children was studied by serum bactericidal assays (SBA). First, isogenic vaccine strains and PorA-identical patient isolates were compared as a target in SBA, to ensure that the vaccine strains are representative for patient isolates. Geometric mean titers (GMTs) in SBA against patient isolates with subtypes P1.5-2,10 and P1.5-1,2-2 after vaccination with HexaMen were generally lower than those against vaccine strains with the same subtype, although the percentage of vaccine responders (> or =4-fold increase in SBA after vaccination) was not affected. Using various P1.7-2,4 patient isolates, GMTs as well as the number of vaccine responders were higher than for the P1.7 2,4 vaccine strain, indicating that the use of the P1.7-2,4 vaccine strain may have underestimated the immunogenicity of this subtype in HexaMen. Secondly, the cross-reactivity of antibodies induced by MonoMen and HexaMen was studied using several patient isolates that differed from the vaccine subtypes by having minor antigenic variants of one variable region (VR), by having a completely different VR or by having a different combination of VRs. MonoMen induced P1.4-specific antibodies that were cross-reactive with P1.4 variants P1.4-1 and P1.4-3. HexaMen induced a broader cross-reactive antibody response against various patient isolates with one VR identical to a vaccine subtype or a combination of VRs included in HexaMen. Cross-reactivity, measured by a fourfold increase in SBA after vaccination, against these strains ranged from 23 to 92% depending on the subtype of the tested strain and was directed against both VR1 and VR2. The extended cross-reactivity of vaccinee sera induced by HexaMen against antigenic variants has important favorable implications for meningococcal B OMV vaccine coverage. PMID- 12654778 TI - Virulence, immunogenicity, and protective efficacy of two recombinant Mycobacterium bovis bacillus Calmette-Guerin strains expressing the antigen ESAT 6 from Mycobacterium tuberculosis. AB - We constructed two recombinant Mycobacterium bovis BCG (rBCG) strains expressing ESAT-6 of Mycobacterium tuberculosis, named rBCG-1 and rBCG-2. rBCG-1 contained the ESAT-6 gene linked to BCG hsp60 and expressed a fusion protein, while rBCG-2, with a secretory sequence, could secret ESAT-6 into the culture medium. There was no evidence for increased virulence of the two rBCG strains when we made a comparison between them and BCG with regard to organ bacterial loads, lung histology, and survival time. rBCG-1 induced significantly higher specific antibody titers and stronger cellular immune response than BCG, whereas rBCG-2 had immunogenicity similar to that of the parental BCG strain. Both rBCG-1 and rBCG-2 conferred marked protection against M. tuberculosis infection, yet in terms of protective efficacy, they showed no significant improvements upon conventional BCG vaccine. PMID- 12654779 TI - Surfactant protein D-mediated aggregation of Pneumocystis carinii impairs phagocytosis by alveolar macrophages. AB - Pneumocystis carinii remains an important and potentially fatal cause of opportunistic pneumonia. Animal studies reveal that substantial quantities of surfactant protein D (SP-D) accumulate in the airspaces during P. carinii pneumonia and are particularly abundant in aggregates of organisms. Due to the multimeric structure of SP-D, we hypothesized that SP-D mediates aggregation of the organism. From previous clinical studies it is known that aggregated organisms are conspicuous in sections of lung tissue and bronchoalveolar lavage (BAL) fluids of humans with active P. carinii pneumonia. Herein, we observe that SP-D levels increased at least fourfold in BAL fluids of patients with P. carinii pneumonia. Next, a spectrophotometric sedimentation assay was developed to assess the aggregation of P. carinii in vitro by SP-D. P. carinii organisms were first stripped with glutathione to remove bound SP-D and subsequently incubated in the presence of SP-D and 2 mM calcium. P. carinii incubated with natural SP-D (10 micro g/ml) containing dodecamers and higher-order forms exhibited aggregation and enhanced sedimentation compared to that of glutathione-stripped P. carinii. Aggregation was also enhanced by the concentrated supernatant of rat BAL fluid, and this effect was abolished by the selective removal of SP-D from the lavage fluid. P. carinii aggregation was reduced by maltose, mannose, and EDTA, consistent with the role of the SP-D C-type lectin domain (CRD) in the aggregation event. Comparisons of different molecular forms of SP-D showed that dodecamers-but not trimeric subunits-mediate optimal aggregation of P. carinii. Aggregation of P. carinii by SP-D was shown to be responsible for the impaired phagocytosis of the organisms by alveolar macrophages. Thus, SP-D-mediated aggregation of P. carinii may represent one means by which the organism avoids elimination by the host. PMID- 12654780 TI - A new vaccine against tuberculosis affords greater survival after challenge than the current vaccine in the guinea pig model of pulmonary tuberculosis. AB - Tuberculosis (TB) remains an enormous global health problem, and a new vaccine against TB more potent than the current inadequate vaccine, Mycobacterium bovis BCG, is urgently needed. We describe a recombinant BCG vaccine (rBCG30) expressing and secreting the 30-kDa major secretory protein of Mycobacterium tuberculosis, the primary causative agent of TB, that affords greater survival after challenge than parental BCG in the highly demanding guinea pig model of pulmonary TB. Animals immunized with rBCG30 and then challenged by aerosol with a highly virulent strain of M. tuberculosis survived significantly longer than animals immunized with conventional BCG. The parental and recombinant vaccine strains are comparably avirulent in guinea pigs, as they display a similar pattern of growth and clearance in the lung, spleen, and regional lymph nodes. The pMTB30 plasmid encoding the 30-kDa protein is neither self-transmissible nor mobilizable to other bacteria, including mycobacteria. The pMTB30 plasmid can be stably maintained in Escherichia coli but is expressed only in mycobacteria. The recombinant and parental strains are sensitive to the same antimycobacterial antibiotics. rBCG30, the first vaccine against TB more potent than nearly century old BCG, is being readied for human clinical trials. PMID- 12654781 TI - Cloning and expression of the Escherichia coli K1 outer membrane protein A receptor, a gp96 homologue. AB - Escherichia coli is one of the most common gram-negative bacteria that cause meningitis in neonates. Our previous studies have shown that outer membrane protein A (OmpA) of E. coli interacts with a 95-kDa human brain microvascular endothelial cell (HBMEC) glycoprotein, Ecgp, for invasion. Here, we report the identification of a gene that encodes Ecgp by screening of an HBMEC cDNA expression library as well as by 5' rapid amplification of cDNA ends. The sequence of the Ecgp gene shows that it is highly similar to gp96, a tumor rejection antigen-1, and contains an endoplasmic reticulum retention signal, KDEL. Overexpression of either Ecgp or gp96 in both HBMECs and CHO cells increases E. coli binding and invasion. We further show that Ecgp gene transfected HBMECs express Ecgp on the cell surface despite the presence of the KDEL motif. Northern blot analysis of total RNA from various eukaryotic cells indicates that Ecgp is significantly expressed in HBMECs. Recombinant His-tagged Ecgp blocked E. coli invasion efficiently by binding directly to the bacteria. These results suggest that OmpA of E. coli K1 interacts with a gp96-like molecule on HBMECs for invasion. PMID- 12654783 TI - Expression of multiple outer membrane protein sequence variants from a single genomic locus of Anaplasma phagocytophilum. AB - Anaplasma phagocytophilum is the causative agent of an emerging tick-borne zoonosis in the United States and Europe. The organism causes a febrile illness accompanied by other nonspecific symptoms and can be fatal, especially if treatment is delayed. Persistence of A. phagocytophilum within mammalian reservoir hosts is important for ensuring continued disease transmission. In the related organism Anaplasma marginale, persistence is associated with antigenic variation of the immunoprotective outer membrane protein MSP2. Extensive diversity of MSP2 is achieved by combinatorial gene conversion of a genomic expression site by truncated pseudogenes. The major outer membrane protein of A. phagocytophilum, MSP2(P44), is homologous to MSP2 of A. marginale, has a similar organization of conserved and variable regions, and is also encoded by a multigene family containing some truncated gene copies. This suggests that the two organisms could use similar mechanisms to generate diversity in outer membrane proteins from their small genomes. We define here a genomic expression site for MSP2(P44) in A. phagocytophilum. As in A. marginale, the msp2(p44) gene in this expression site is polymorphic in all populations of organisms we have examined, whether organisms are obtained from in vitro culture in human HL-60 cells, from culture in the tick cell line ISE6, or from infected human blood. Changes in culture conditions were found to favor the growth and predominance of certain msp2(p44) variants. Insertions, deletions, and substitutions in the region of the genomic expression site encoding the central hypervariable region matched sequence polymorphisms in msp2(p44) mRNA. These data suggest that, similarly to A. marginale, A. phagocytophilum uses combinatorial mechanisms to generate a large array of outer membrane protein variants. Such gene polymorphism has profound implications for the design of vaccines, diagnostic tests, and therapy. PMID- 12654782 TI - Profiling of temperature-induced changes in Borrelia burgdorferi gene expression by using whole genome arrays. AB - Borrelia burgdorferi is the etiologic agent of Lyme disease, the most prevalent arthropod-borne disease in the United States. The genome of the type strain, B31, consists of a 910,725-bp linear chromosome and 21 linear and circular plasmids comprising 610,694 bp. During its life cycle, the spirochete exists in distinctly different environments, cycling between a tick vector and a mammalian host. Temperature is one environmental factor known to affect B. burgdorferi gene expression. To identify temperature-responsive genes, genome arrays containing 1,662 putative B. burgdorferi open reading frames (ORFs) were prepared on nylon membranes and employed to assess gene expression in B. burgdorferi B31 grown at 23 and 35 degrees C. Differences in expression of more than 3.5 orders of magnitude could be readily discerned and quantitated. At least minimal expression from 91% of the arrayed ORFs could be detected. A total of 215 ORFs were differentially expressed at the two temperatures; 133 were expressed at significantly greater levels at 35 degrees C, and 82 were more significantly expressed at 23 degrees C. Of these 215 ORFs, 134 are characterized as genes of unknown function. One hundred thirty-six (63%) of the differentially expressed genes are plasmid encoded. Of particular interest is plasmid lp54 which contains 76 annotated putative genes; 31 of these exhibit temperature-regulated expression. These findings underscore the important role plasmid-encoded genes may play in adjustment of B. burgdorferi to growth under diverse environmental conditions. PMID- 12654784 TI - Role of antibodies in immunity to Bordetella infections. AB - The persistence of Bordetella pertussis and B. parapertussis within vaccinated populations and the reemergence of associated disease highlight the need to better understand protective immunity. The present study examined host immunity to bordetellae and addressed potential concerns about the mouse model by using a comparative approach including the closely related mouse pathogen B. bronchiseptica. As previously observed with B. pertussis, all three organisms persisted throughout the respiratory tracts of B-cell-deficient mice, indicating that B cells are required for bacterial clearance. However, adoptively transferred antibodies rapidly cleared B. bronchiseptica but not human pathogens. These results obtained with the mouse model are consistent with human clinical observations, including the lack of correlation between antibody titers and protection, as well as the limited efficacy of intravenous immunoglobulin treatments against human disease. Together, this evidence suggests that the mouse model accurately reflects substantial differences between immunities to these organisms. Although both B. pertussis and B. parapertussis are more closely related to B. bronchiseptica than they are to each other, they share the ability to resist rapid clearance from the lower respiratory tract by adoptively transferred antibodies, an adaptation that correlates with their emergence as human pathogens that circulate within vaccinated populations. PMID- 12654785 TI - Differential modulation by Ca2+ of type III secretion of diffusely adhering enteropathogenic Escherichia coli. AB - Enteropathogenic Escherichia coli (EPEC) strains are a common cause of persistent diarrhea among infants, primarily in developing countries. The pathogenicity of EPEC is associated with the expression and secretion of bacterial proteins encoded by the chromosomal locus of enterocyte effacement (LEE). The LEE-encoded type III-secreted proteins EspA, EspB, and EspD are part of a molecular syringe, which is used by EPEC to translocate effector proteins directly into the cytoplasm of host cells. The type III-secreted translocated intimin receptor (Tir) protein is thought to be delivered by an Esp-dependent mechanism into the host cell, and this is followed by insertion into the host plasma membrane, where the protein serves as the receptor for intimin, an afimbrial bacterial adhesin. Type III secretion is subject to environmental regulation, and secretion can be induced in vitro by growing bacteria in cell culture medium. In this study we found that Ca(2+) is involved in the regulation of type III secretion both in classical locally adherent EPEC and in atypical diffusely adherent EPEC. Interestingly, we observed contrasting secretion of Esp proteins and Tir in response to Ca(2+). While the secretion of Tir is clearly enhanced and the protein is integrated into HeLa membranes under calcium chelation conditions, Esp secretion is strongly reduced under these conditions. These data suggest that under Ca(2+)-depleted conditions Tir might be secreted into the medium and integrated into host membranes by an Esp-independent mechanism, without the need for a functional type III translocation machinery. PMID- 12654787 TI - Cholera toxin and its B subunit promote dendritic cell vaccination with different influences on Th1 and Th2 development. AB - Cholera toxin (CT) is a strong mucosal adjuvant for codelivered antigens, whereas its nontoxic B subunit (CTB) is an efficient mucosal carrier molecule for the generation of immune responses to linked antigens. We investigated the effects of CT and CTB on the immunogenicity of in vitro-treated antigen-pulsed dendritic cells (DC) following intravenous injection into mice. Prior to infusion, DC were pulsed for 90 min with either free ovalbumin (OVA), OVA mixed with CT or CTB, or chemical conjugates of OVA with CT and CTB (OVA-CT and OVA-CTB). DC pulsed with OVA or with OVA and CTB gave rise to modest antibody and T-cell responses. Conjugation of OVA with CTB enhanced both the subsequent B-cell and T-cell responses to OVA and preferentially induced Th2 responses. CT was shown to be a strong adjuvant when it was coadministered to DC with OVA and was even stronger when it was coadministered with OVA-CTB and primed for a mixed Th1-Th2 response. The antibody and T-cell responses were further enhanced if OVA was coupled to CT, implying that CT can utilize a combined carrier and adjuvant function vis-a-vis linked antigens for DC vaccination. The immunopotentiating capacity of CT- and CTB-linked antigen was associated with both upregulated secretion of interleukin 1beta by the pulsed DC and increased expression of CD80 and CD86 on the DC surface. These results imply that CT and CTB can be used to both markedly increase and partially direct the DC vaccine-induced immune response with respect to Th1 and Th2 responses, which has obvious implications for DC-based vaccine development. PMID- 12654786 TI - Modification in the ppk gene of Helicobacter pylori during single and multiple experimental murine infections. AB - The bacterial pathogen Helicobacter pylori is highly adapted to the human stomach, and a high level of polymorphism is observed among clinical isolates. This polymorphism may be the consequence of adaptive changes during colonization, making a strain better able to survive, to evade the immune system, and to provoke a chronic infection. To investigate the mechanisms involved in the acquisition of diversity in H. pylori, mouse models of single infections, coinfections, and superinfections were developed. These experimental infections were conducted with strain SS1, well known to be mouse adapted, and with two strains freshly isolated from infected patients: Hp141 and Hp145. Genetic modifications occurring in these strains were studied over time by comparing randomly selected colonies of the emerging strains to those of the infecting strains by using randomly amplified polymorphic DNA fingerprinting with six different primers and by using PCR to amplify the vacA and cagA genes. We showed that, regardless of the number of infecting strains, only one emerged from the animals and that the establishment of a first strain thwarted the implantation of a second strain. During both a single infection and a coinfection with SS1, Hp141 was replaced by a genetic variant (Hp141v) that overcame SS1 in coinfection experiments. Hp141v exhibited a deletion of a 102-bp repeated sequence within the ppk gene, which encodes polyphosphate kinase (PPK), an enzyme involved in the physiological adaptation of the microbial cell to nutritional and environmental stresses. The deletion led to higher enzymatic activity of PPK, and the variant exhibited a better capacity to colonize mice. Considering that the modified gene is known to be involved in adaptation to a new environment, our results are consistent with an adaptive change in strain Hp141 and suggest that PPK is an important virulence factor in H. pylori. PMID- 12654788 TI - Induction of protective immunity to Listeria monocytogenes with dendritic cells retrovirally transduced with a cytotoxic T lymphocyte epitope minigene. AB - In the present study, we developed a cytotoxic T lymphocyte (CTL) epitope minigene-transduced dendritic cell (DC)-based vaccine against Listeria monocytogenes. Murine bone marrow-derived DCs were retrovirally transduced with a minigene for listeriolysin O (LLO) 91-99, a dominant CTL epitope of L. monocytogenes, and were injected into BALB/c mice intravenously. We found that the DC vaccine was capable of generating peptide-specific CD8+ T cells exhibiting LLO 91-99-specific cytotoxic activity and gamma interferon production, leading to induction of protective immunity to the bacterium. Furthermore, we demonstrated that the retrovirally transduced DC vaccine was more effective than a CTL epitope peptide-pulsed DC vaccine and a minigene DNA vaccine for eliciting antilisterial immunity. These results provide an alternative strategy in which retrovirally transduced DCs are used to design vaccines against intracellular pathogens. PMID- 12654789 TI - Helicobacter pylori-specific CD4+ CD25high regulatory T cells suppress memory T cell responses to H. pylori in infected individuals. AB - Helicobacter pylori colonizes the gastric and duodenal mucosa. The infection normally persists for life and causes peptic ulcers and gastric cancer in a subset of infected individuals. We hypothesized that the inability to clear the infection may be a consequence of H. pylori-specific regulatory T cells that actively suppress T-cell responses. Therefore, we characterized the T-cell responses to H. pylori in H. pylori-infected individuals without any subjective symptoms and in uninfected control subjects and investigated the role of regulatory CD4+ CD25(high) T cells during infection. The stimulation of CD4+ peripheral blood T cells with monocyte-derived dendritic cells pulsed with a membrane preparation of H. pylori resulted in proliferation and gamma interferon production in both infected and uninfected individuals. Sorted memory cells from infected individuals responded less than cells from uninfected subjects, and the unresponsiveness could be abolished by depletion of CD4+ CD25(high) regulatory T cells or the addition of interleukin 2. Furthermore, CD4+ CD25(high) T cells suppressed H. pylori-induced responses in cocultures with CD25(low/-) cells. Tetanus toxoid induced comparable responses in memory cells from infected and uninfected individuals in both the presence and the absence of regulatory T cells, suggesting that the suppression was H. pylori specific. In conclusion, we have shown that H. pylori-infected individuals have impaired memory CD4+ T-cell responses to H. pylori that are linked to the presence of H. pylori-specific regulatory T cells that actively suppress the responses. PMID- 12654790 TI - Differential effects of control and antigen-specific T cells on intracellular mycobacterial growth. AB - We investigated the effects of peripheral blood mononuclear cells expanded with irrelevant control and mycobacterial antigens on the intracellular growth of Mycobacterium bovis bacillus Calmette-Guerin (BCG) in human macrophages. More than 90% of the cells present after 1 week of in vitro expansion were CD3(+). T cells were expanded from purified protein derivative-negative controls, persons with latent tuberculosis, and BCG-vaccinated individuals. T cells expanded with nonmycobacterial antigens enhanced the intracellular growth of BCG in suboptimal cultures of macrophages. T cells expanded with live BCG or lysates of Mycobacterium tuberculosis directly inhibited intracellular BCG. Recent intradermal BCG vaccination significantly enhanced the inhibitory activity of T cells expanded with mycobacterial antigens (P < 0.02), consistent with the induction of memory-immune inhibitory T-cell responses. Selected mycobacterial antigens (Mtb41 > lipoarabinomannan > 38kd > Ag85B > Mtb39) expanded inhibitory T cells, demonstrating the involvement of antigen-specific T cells in intracellular BCG inhibition. We studied the T-cell subsets and molecular mechanisms involved in the memory-immune inhibition of intracellular BCG. Mycobacteria-specific gammadelta T cells were the most potent inhibitors of intracellular BCG growth. Direct contact between T cells and macrophages was necessary for the BCG growth enhancing and inhibitory activities mediated by control and mycobacteria-specific T cells, respectively. Increases in tumor necrosis factor alpha, interleukin-6, transforming growth factor beta, and vascular endothelial growth factor mRNA expression were associated with the enhancement of intracellular BCG growth. Increases in gamma interferon, FAS, FAS ligand, perforin, granzyme, and granulysin mRNA expression were associated with intracellular BCG inhibition. These culture systems provide in vitro models for studying the opposing T-cell mechanisms involved in mycobacterial survival and protective host immunity. PMID- 12654793 TI - Genetic organization and iron-responsive regulation of the Brucella abortus 2,3 dihydroxybenzoic acid biosynthesis operon, a cluster of genes required for wild type virulence in pregnant cattle. AB - Brucella abortus reportedly produces the monocatechol siderophore 2,3 dihydroxybenzoic acid (2,3-DHBA) in response to iron limitation. Nucleotide sequence analysis of the cloned DHBA biosynthesis locus from virulent B. abortus 2308 and genetic complementation of defined Escherichia coli mutants were used to identify the B. abortus genes (designated dhbC, -B, and -A) responsible for synthesis of this siderophore. Reverse transcriptase PCR analysis of total RNA with dhb-specific primers demonstrated that dhbC, -B, and -A are transcribed as components of an operon, together with dhbE, a functional homolog of the Escherichia coli entE gene. Homologs of the E. coli entD and Vibrio cholerae vibH genes were also detected in the flanking regions immediately adjacent to the B. abortus dhbCEBA operon, suggesting that B. abortus has the genetic capacity to produce a more complex 2,3-DHBA-based siderophore. Slot blot hybridization experiments and primer extension analysis showed that transcription of the B. abortus dhbCEBA operon originates from two iron-regulated promoters located upstream of dhbC. Consistent with their iron-dependent regulation, both of the dhbCEBA promoter sequences contain typical consensus Fur-binding motifs. Although previously published studies have shown that 2,3-DHBA production is not required for the establishment and maintenance of chronic spleen infection by B. abortus in mice, experimental infection of pregnant cattle with the B. abortus dhbC mutant BHB1 clearly showed that production of this siderophore is essential for wild-type virulence in the natural ruminant host. PMID- 12654791 TI - Afa/Dr diffusely adhering Escherichia coli infection in T84 cell monolayers induces increased neutrophil transepithelial migration, which in turn promotes cytokine-dependent upregulation of decay-accelerating factor (CD55), the receptor for Afa/Dr adhesins. AB - Ulcerative colitis and Crohn's disease are inflammatory bowel diseases thought to involve strains of Escherichia coli. We report here that two wild-type Afa/Dr diffusely adhering E. coli (DAEC) strains, C1845 and IH11128, which harbor the fimbrial F1845 adhesin and the Dr hemagglutinin, respectively, and the E. coli laboratory strain HB101, transformed with the pSSS1 plasmid to produce Afa/Dr F1845 adhesin, all induced interleukin-8 (IL-8) production and transepithelial migration of polymorphonuclear leukocytes (PMNL) in polarized monolayers of the human intestinal cell line T84 grown on semipermeable filters. We observed that after PMNL migration, expression of decay-accelerating factor (DAF, or CD55), the brush border-associated receptor for Afa/Dr adhesins, was strongly enhanced, increasing the adhesion of Afa/Dr DAEC bacteria. When examining the mechanism by which DAF expression was enhanced, we observed that the PMNL transepithelial migration induced epithelial synthesis of tumor necrosis factor alpha and IL 1beta, which in turn promoted the upregulation of DAF. PMID- 12654794 TI - Attenuated Yersinia enterocolitica mutant strains exhibit differential virulence in cytokine-deficient mice: implications for the development of novel live carrier vaccines. AB - Yersinia enterocolitica mutant strains, including mutants deficient in the chaperone SycH resulting in a functional deficiency in tyrosine phosphatase (YopH), Mn-cofactored superoxide dismutase (SodA), iron-repressive protein 1 (IRP 1), and Yersinia adhesin A (YadA), were demonstrated to be highly attenuated in wild-type C57BL/6 mice. TNFRp55(-/-), IL-12p40(-/-), and IL-18(-/-) mutant mice, in which the Yersinia wild-type strain causes severe systemic infections, were used to investigate whether these Yersinia mutant strains would be attenuated in immunodeficient hosts. A plasmid-cured Yersinia mutant strain was unable to colonize any of the mutant mice tested. A SycH-deficient mutant strain colonized intestinal tissues of these mice but was attenuated for systemic infection in all of the mutant mice. Both YadA- and Irp-1-deficient Yersinia mutants were still attenuated in IL-12(-/-) and IL-18(-/-) mice but were pathogenic in TNFRp55(-/-) mice. By contrast, a Yersinia sodA mutant was highly pathogenic for TNFRp55(-/-) and IL-12p40(-/-) mice while interleukin-18 (IL-18) was dispensable. This finding demonstrates that certain virulence factors enable yersiniae to compete with distinct cytokine-dependent host defense mechanisms. Moreover, while gamma interferon mRNA expression did not reflect protective host responses in cytokine deficient mice, IL-10 expression coincided with a heavy splenic bacterial load and was associated with progressive infection courses. We can thus segregate minor (SodA), intermediate (YadA and IRP-1), and major (YopH) virulence factors of Y. enterocolitica. Finally, we demonstrate that, even in immunocompromised hosts, Yersinia sycH and, with some restrictions, irp-1 mutants may be suitable for use as live carrier vaccines. PMID- 12654792 TI - Expression of Clostridium difficile toxins A and B and their sigma factor TcdD is controlled by temperature. AB - Growth temperature was found to control the expression of toxins A and B in Clostridium difficile VPI 10463, with a maximum at 37 degrees C and low levels at 22 and 42 degrees C in both peptone yeast (PY) and defined media. The up regulation of toxin A and B mRNA and protein levels upon temperature upshift from 22 to 37 degrees C followed the same kinetics, showing that temperature control occurred at the level of transcription. Experiments with Clostridium perfringens using gusA as a reporter gene demonstrated that both toxin gene promoters were temperature controlled and that their high activity at 37 degrees C was dependent on the alternative sigma factor TcdD. Furthermore, tcdD was found to be autoinduced at 37 degrees C. Glucose down-regulated all these responses in the C. perfringens constructs, similar to its impact on toxin production in C. difficile PY broth cultures. C. difficile proteins induced at 37 degrees C and thus coregulated with the toxins by temperature were demonstrated by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified as enzymes involved in butyric acid production and as electron carriers in oxidation reduction reactions. The regulation of toxin production in C. difficile by temperature is a novel finding apparently reflecting an adaptation of the expression of its virulence to mammalian hosts. PMID- 12654795 TI - Pneumococcal behavior and host responses during bronchopneumonia are affected differently by the cytolytic and complement-activating activities of pneumolysin. AB - Pneumolysin, a multifunctional toxin produced by all clinical isolates of Streptococcus pneumoniae, is strongly implicated in the pathogenesis of pneumococcal bronchopneumonia and septicemia. Using isogenic mutant strains, we examined the effect of deletion of the cytotoxic activity or complement activating activity of pneumolysin on bacterial growth in lungs and blood, histological changes in infected lung tissue, and the pattern of inflammatory cell recruitment. Both of the activities of pneumolysin contributed to the pathology in the lungs, as well as the timing of the onset of bacteremia. Histological changes in the lungs were delayed after infection with either mutant compared to the changes seen after infection with the wild-type pneumococcus. The complement-activating activity of pneumolysin affected the accumulation of T cells, whereas the toxin's cytolytic activity influenced neutrophil recruitment into lung tissue. PMID- 12654797 TI - Characterization of Cryptosporidium meleagridis of human origin passaged through different host species. AB - Cryptosporidium meleagridis, a protozoon first observed in turkeys, has been linked by several investigators to cryptosporidiosis in humans. C. meleagridis is the only known Cryptosporidium species that infects both avian and mammalian species. We describe the successful propagation of C. meleagridis (isolate TU1867), originally purified from a patient with diarrhea, in laboratory animals including chickens, mice, piglets, and calves. TU1867 was readily transmitted from one animal host to another, maintaining genetic homogeneity and stability. The rate of infectivity and virulence of TU1867 for the mammalian species were similar to those of Cryptosporidium parvum. Laboratory propagation of genetically and phenotypically stable and well-characterized reference isolates, representing various Cryptosporidium species, particularly those infectious to humans, will improve considerably the spectrum and quality of laboratory and field investigations on this medically important protozoa. PMID- 12654796 TI - Protective efficacy of anti-Helicobacter pylori immunity following systemic immunization of neonatal mice. AB - Helicobacter pylori infection of the gastric mucosa is a significant cause of morbidity and mortality because of its etiologic role in symptomatic gastritis, peptic ulcer disease, and gastric adenocarcinoma. Infection occurs in young children; therefore, a prophylactic vaccine would have to be administered within the first year of life, a period thought to be immunologically privileged. We investigated vaccine formulations administered by different routes to confer protective anti-H. pylori immunity in neonatal mice. Neonatal mice immunized with a single dose of vaccine in complete Freund's adjuvant (CFA) generated antigen specific gamma interferon-, interleukin-2 (IL-2)-, IL-4-, and IL-5-secreting T cells in numbers similar to those in immunized adult mice, while vaccine administered to neonates in incomplete Freund's adjuvant (IFA) induced such cells in reduced numbers compared to those in adult mice. Both IFA and CFA, however, provided partial protection from a challenge with infectious H. pylori when the vaccine was administered subcutaneously. Neonatal immunized mice also had reduced bacterial loads when immunized intraperitoneally with CFA. In all cases, protection was equivalent to that achieved when adult counterparts were immunized. These studies suggest that an efficacious vaccine might be successfully administered to very young children to prevent perinatal infection of H. pylori. PMID- 12654798 TI - Repeat sequences in block 2 of Plasmodium falciparum merozoite surface protein 1 are targets of antibodies associated with protection from malaria. AB - Human antibodies to the block 2 region of Plasmodium falciparum merozoite surface protein 1 (MSP1) are associated with a reduced prospective risk of clinical malaria. Block 2 is highly polymorphic, but all known alleles can be grouped into three major types. Two of these types (the K1-like and MAD20-like types) contain type-specific sequences (found in all alleles of a particular type) that flank polymorphic tripeptide repeats. These repeats contain both type-specific and subtype-specific sequences. To evaluate the antibody recognition of these parts of block 2, a new panel of six recombinant proteins was used (fused type-specific flanking sequences and two representative repeat sequences for each of the K1 like and MAD20-like types separately). Extensive testing of these antigens and full-length block 2 antigens showed that human serum immunoglobulin G antibodies induced by infection can recognize (i) type-specific epitopes in the repeats, (ii) subtype-specific epitopes in the repeats, or (iii) type-specific epitopes in flanking sequences. A large prospective study in The Gambia showed that antibodies to the repeats are strongly associated with protection from clinical malaria. The results are important for design of a vaccine to induce protective antibodies, and they address hypotheses about repeat sequences in malaria antigens. PMID- 12654799 TI - Inactivation of the Moraxella catarrhalis 7169 ferric uptake regulator increases susceptibility to the bactericidal activity of normal human sera. AB - Moraxella catarrhalis is a strict human pathogen and a significant cause of respiratory disease and otitis media. In direct response to these infections, research efforts have focused primarily on the identification of potential vaccine targets. The general biology of M. catarrhalis, however, including the mechanisms utilized to survive in the human host, remains poorly understood. Previous work has demonstrated that M. catarrhalis expresses iron-repressible proteins, suggesting the presence of iron acquisition systems under the control of a ferric uptake regulator (Fur). In this study M. catarrhalis fur has been cloned and sequenced from strain 7169. A deletion-insertion mutation of 7169 fur resulted in upregulation of iron-repressible outer membrane proteins in the absence and presence of iron. This mutant strain, 7169fur1, was significantly more sensitive to the bactericidal activity of normal human serum than the resistant wild-type strain. These data suggest that constitutive expression of iron-regulated proteins may provide multiple targets for human antibodies. In addition, the 7169 fur mutant provides an important tool for further investigation of the iron acquisition mechanisms utilized by M. catarrhalis. PMID- 12654800 TI - Development and evaluation of an improved mouse model of meningococcal colonization. AB - Studies of meningococcal pathogenesis have been severely restricted due to the absence of an adequate animal model. Given the significance of iron in meningococcal pathogenesis, we developed a model of Neisseria meningitidis colonization in outbred adult mice that included daily administration of iron dextran. While receiving iron, the animals were inoculated intranasally with the initial doses of bacterial suspension. Meningococci were recovered from the animals by nasopharyngeal washes. Approximately half of the animals inoculated with 10(7) CFU remained colonized 13 days after the initial bacterial inoculation. The model was further evaluated with genetically defined isogenic serogroup B mutant strains, and the colonization capabilities of the mutants were compared to that of the wild-type parent. A mutant that produces truncated lipooligosaccharide (KDO(2)-lipid A) and a mutant defective in capsule transport were dramatically impaired in colonization. A mutant defective in pilus transport (pilQ) showed moderately impaired colonization. The immunological aspect of the model was also evaluated by challenging mice after immunization with homologous whole-cell meningococci. The immunized mice were protected from colonization of the homologous strain. In this model, long-term meningococcal colonization was maintained, allowing us to study the effects of specific genetic mutation on colonization. In addition, this model allows investigation of the role of active immune response against meningococci. PMID- 12654801 TI - Erythrocyte invasion phenotypes of Plasmodium falciparum in The Gambia. AB - In vitro experimentation with Plasmodium falciparum has determined that a number of different receptor-ligand interactions are involved in the invasion of erythrocytes. Most culture-adapted parasite isolates use a mechanism of invasion that depends primarily on the erythrocyte sialoglycoprotein glycophorin A (GYPA) and erythrocyte-binding antigen 175 (EBA-175) of the parasite blood-stage merozoite. However, a minority of culture-adapted parasites and a majority of Indian field isolates can apparently invade by other means. Here, erythrocyte invasion phenotypes of P. falciparum field isolates in Africa were studied. For 38 Gambian isolates, invasion of neuraminidase-treated and trypsin-treated erythrocytes was inhibited, on average, by more than 60 and 85%, respectively, indicating a high level of dependence on sialic acid and trypsin-sensitive proteins on the erythrocyte surface. These results support the hypothesis that African P. falciparum parasites use GYPA as a primary receptor for invasion. However, the considerable variation among isolates confirms the idea that alternative receptors are also used by many parasites. Three amino acid polymorphisms in the GYPA-binding region of EBA-175 (region II) were not significantly associated with invasion phenotype. There was variation among isolates in the selectivity index (i.e., a statistical tendency toward aggregation or multiple invasions of host erythrocytes), but this variation did not correlate with enzyme-determined invasion phenotype or with eba-175 alleles. Overall, these invasion phenotypes in Africa support a vaccine strategy of inhibiting EBA-175 binding to GYPA but suggest that parasites with alternative phenotypes would be selected for if this strategy were used alone. PMID- 12654802 TI - Role of glutathione in macrophage control of mycobacteria. AB - Reactive oxygen and nitrogen intermediates are important antimicrobial defense mechanisms of macrophages and other phagocytic cells. While reactive nitrogen intermediates have been shown to play an important role in tuberculosis control in the murine system, their role in human disease is not clearly established. Glutathione, a tripeptide and antioxidant, is synthesized at high levels by cells during reactive oxygen intermediate and nitrogen intermediate production. Glutathione has been recently shown to play an important role in apoptosis and to regulate antigen-presenting-cell functions. Glutathione also serves as a carrier molecule for nitric oxide, in the form of S-nitrosoglutathione. Previous work from this laboratory has shown that glutathione and S-nitrosoglutathione are directly toxic to mycobacteria. A mutant strain of Mycobacterium bovis BCG, defective in the transport of small peptides such as glutathione, is resistant to the toxic effect of glutathione and S-nitrosoglutathione. Using the peptide transport mutant as a tool, we investigated the role of glutathione and S nitrosoglutathione in animal and human macrophages in controlling intracellular mycobacterial growth. PMID- 12654803 TI - Novel virulence-associated type II secretion system unique to high-pathogenicity Yersinia enterocolitica. AB - Yersinia enterocolitica strains comprise an important group of bacterial enteropathogens that cause a broad range of gastrointestinal syndromes. Three groups are distinguishable within this bacterial species, namely, the nonpathogenic group (biotype 1A strains), the low-pathogenicity, non-mouse-lethal group (biotypes 2 to 5), and the high-pathogenicity, mouse-lethal group (biotype 1B). To date, the presence of the high-pathogenicity island (HPI), a chromosomal locus that encodes the yersiniabactin system (involved in iron uptake), defines essentially the difference between low-pathogenicity and high-pathogenicity Y. enterocolitica strains, with the low-pathogenicity strains lacking the HPI. Using the powerful tool of representational difference analysis between the nonpathogenic 1A strain, NF-O, and its high-pathogenicity 1B counterpart, WA-314, we have identified a novel type II secretion gene cluster (yts1C-S) occurring exclusively in the high-pathogenicity group. The encoded secreton, designated Yts1 (for Yersinia type II secretion 1) was shown to be important for virulence in mice. A close examination of the almost completed genome sequence of another high-pathogenicity representative, Y. enterocolitica 8081, revealed a second putative type II secretion cluster uniformly distributed among all Y. enterocolitica isolates. This putative species-specific cluster (designated yts2) differed significantly from yts1, while resembling more closely the putative type II cluster present on the genome of Y. pestis. The Yts1 secreton thus appears to have been additionally acquired by the high-pathogenicity assemblage for a virulence-associated function. PMID- 12654804 TI - Oral transfer of adult Ancylostoma ceylanicum hookworms into permissive and nonpermissive host species. AB - Syrian hamsters become anemic and exhibit delayed growth following oral infection with third-stage Ancylostoma ceylanicum hookworm larvae. Here we describe experiments designed to determine the feasibility of adult worm transfer (AWT) between hosts, a technique that would facilitate the specific study of bloodfeeding hookworms in vivo without prior exposure of the host to larva specific antigens, permit the ex vivo manipulation of adult parasites prior to reimplantation, and also allow for cross-species transfer of worms. Weanling hamsters given an oral AWT of 40 or 60 mixed-sex A. ceylanicum worms rapidly developed anemia; in the higher-dose group, hemoglobin levels declined from prechallenge levels by 44% within 4 days following AWT. Long-term survival of transferred worms was demonstrated by recovery of parasites from the intestines 42 days after AWT. AWT hamsters acquired humoral immune responses against soluble adult hookworm extracts and excretory-secretory products that were comparable in magnitude to those of animals given a typical infection with larvae. In AWT experiments employing the nonpermissive murine model, C57BL/6 mice given adult worms rapidly became anemic and lost weight in a manner similar to AWT hamsters. Infection of additional mouse strains demonstrated that while C57BL/10 and CD-1 mice also developed anemia following AWT, BALB/c mice were resistant. The technique of AWT to mice may further our understanding of hookworm pathogenesis by allowing the study of adult hookworm infections in a species with well characterized genetics and an abundance of available reagents. PMID- 12654806 TI - Zonula occludens toxin acts as an adjuvant through different mucosal routes and induces protective immune responses. AB - Zonula occludens toxin (Zot) is produced by Vibrio cholerae and has the ability to increase mucosal permeability by reversibly affecting the structure of tight junctions. Because of this property, Zot is a promising tool for mucosal drug and antigen (Ag) delivery. Here we show that Zot acts as a mucosal adjuvant to induce long-lasting and protective immune responses upon mucosal immunization of mice. Indeed, the intranasal delivery of ovalbumin with two different recombinant forms of Zot in BALB/c mice resulted in high Ag-specific serum immunoglobulin G titers that were maintained over the course of a year. Moreover, His-Zot induced humoral and cell-mediated responses to tetanus toxoid in C57BL/6 mice and protected the mice against a systemic challenge with tetanus toxin. In addition, we found that Zot also acts as an adjuvant through the intrarectal route and that it has very low immunogenicity compared to the adjuvant Escherichia coli heat-labile enterotoxin. Finally, by using an octapeptide representing the putative binding site of Zot and of its endogenous analogue zonulin, we provide evidence that Zot may bind a mucosal receptor on nasal mucosa and may mimic an endogenous regulator of tight junctions to deliver Ags in the submucosa. In conclusion, Zot is a novel and effective mucosal adjuvant that may be useful for the development of mucosal vaccines. PMID- 12654805 TI - Intranasal immunization with recombinant Lactococcus lactis secreting murine interleukin-12 enhances antigen-specific Th1 cytokine production. AB - Interleukin-12 (IL-12), a heterodimeric cytokine, plays an important role in cellular immunity to several bacterial, viral, and parasitic infections and has adjuvant activity when it is codelivered with DNA vaccines. IL-12 has also been used with success in cancer immunotherapy treatments. However, systemic IL-12 therapy has been limited by high levels of toxicity. We describe here inducible expression and secretion of IL-12 in the food-grade lactic acid bacterium Lactococcus lactis. IL-12 was expressed as two separate polypeptides (p35-p40) or as a single recombinant polypeptide (scIL-12). The biological activity of IL-12 produced by the recombinant L. lactis strain was confirmed in vitro by its ability to induce gamma interferon (IFN-gamma) production by mouse splenocytes. Local administration of IL-12-producing strains at the intranasal mucosal surface resulted in IFN-gamma production in mice. The activity was greater with the single polypeptide scIL-12. An antigen-specific cellular response (i.e., secretion of Th1 cytokines, IL-2, and IFN-gamma) elicited by a recombinant L. lactis strain displaying a cell wall-anchored human papillomavirus type 16 E7 antigen was dramatically increased by coadministration with an L. lactis strain secreting IL-12 protein. Our data show that IL-12 is produced and secreted in an active form by L. lactis and that the strategy which we describe can be used to enhance an antigen-specific immune response and to stimulate local mucosal immunity. PMID- 12654807 TI - M type 1 and 3 group A streptococci stimulate tissue factor-mediated procoagulant activity in human monocytes and endothelial cells. AB - Streptococcal toxic shock syndrome (StrepTSS) is an invasive infection characterized by marked coagulopathy, multiple organ failure, and rapid tissue destruction and is strongly associated with M type 1 and 3 group A streptococci (GAS). Initiation of the coagulation cascade with formation of microvascular thrombi contributes to multiple organ failure in human cases of gram-negative bacteremia; however, little is known regarding the mechanism of coagulopathy in StrepTSS. Thus, we investigated the abilities of several strains of M type 1 and 3 GAS isolated from human cases of StrepTSS to stimulate production of tissue factor (TF), the principal initiator of coagulation in vivo. Washed, killed M type 1 and 3 GAS, but not M type 6 GAS, elicited high-level TF-mediated procoagulant activity from both isolated human monocytes and cultured human umbilical vein endothelial cells. M type 1 GAS consistently elicited higher levels of TF from monocytes than did M type 3 GAS. GAS-induced TF synthesis in monocytes did not correlate with production of tumor necrosis factor alpha or interleukin-8. Conversely, M type 3 GAS were consistently more potent than M type 1 GAS in stimulating endothelial cell TF synthesis. These results demonstrate that (i) M type 1 and 3 strains of GAS are potent inducers of TF synthesis, (ii) GAS-induced TF synthesis is not simply an epiphenomenon of cytokine generation, and (iii) induction of TF in endothelial cells and monocytes may be M type specific. In total, these findings suggest that a novel interaction between GAS and host cells contributes to the observed coagulopathy in StrepTSS. PMID- 12654809 TI - Contribution of the Shigella flexneri Sit, Iuc, and Feo iron acquisition systems to iron acquisition in vitro and in cultured cells. AB - Shigella flexneri possesses multiple iron acquisition systems, including proteins involved in the synthesis and uptake of siderophores and the Feo system for ferrous iron utilization. We identified an additional S. flexneri putative iron transport gene, sitA, in a screen for S. flexneri genes that are induced in the eukaryotic intracellular environment. sitA was present in all Shigella species and in most enteroinvasive Escherichia coli strains but not in any other E. coli isolates tested. The sit locus consists of four genes encoding a potential ABC transport system. The deduced amino acid sequence of the S. flexneri sit locus was homologous to the Salmonella enterica serovar Typhimurium Sit and Yersinia pestis Yfe systems, which mediate both manganese and iron transport. The S. flexneri sit promoter was repressed by either iron or manganese, and the iron repression was partially dependent upon Fur. A sitA::cam mutation was constructed in S. flexneri. The sitA mutant showed reduced growth, relative to the wild type, in Luria broth containing an iron chelator but formed wild-type plaques on Henle cell monolayers, indicating that the sitA mutant was able to acquire iron and/or manganese in the host cell. However, mutants defective in two of these iron acquisition systems (sitA iucD, sitA feoB, and feoB iucD) formed slightly smaller plaques on Henle cell monolayers. A strain carrying mutations in sitA, feoB, and iucD did not form plaques on Henle cell monolayers. PMID- 12654810 TI - The Apa protein of Mycobacterium tuberculosis stimulates gamma interferon secreting CD4+ and CD8+ T cells from purified protein derivative-positive individuals and affords protection in a guinea pig model. AB - The search to identify Mycobacterium tuberculosis antigens capable of conferring protective immunity against tuberculosis has received a boost owing to the resurgence of tuberculosis over the past two decades. It has long been recognized that lymphoid cells are required for protection against M. tuberculosis. While traditionally the CD4(+) populations of T cells were believed to predominantly serve this protective function, a pivotal role for CD8(+) T cells in this task has been increasingly appreciated. We show that the 50- to 55-kDa Apa protein, specified by the Rv1860 gene of M. tuberculosis, can elicit both lymphoproliferative response and gamma interferon (IFN-gamma) production from peripheral blood mononuclear cells (PBMC) of purified protein derivative (PPD) positive individuals, with significant differences recorded in the levels of responsiveness between PPD-positive healthy controls and pulmonary tuberculosis patients. Flow cytometric analysis of whole blood stimulated with the recombinant Apa protein revealed a sizeable proportion of CD8(+) T cells in addition to CD4(+) T cells contributing to IFN-gamma secretion. PBMC responding to the Apa protein produced no interleukin-4, revealing a Th1 phenotype. A DNA vaccine and a poxvirus recombinant expressing the Apa protein were constructed and tested for their ability to protect immunized guinea pigs against a challenge dose of virulent M. tuberculosis. Although the DNA vaccine afforded little protection, the poxvirus recombinant boost after DNA vaccine priming conferred a significant level of protective immunity, bringing about a considerable reduction in mycobacterial counts from the challenge bacilli in spleens of immunized guinea pigs, a result comparable to that achieved by BCG vaccination. PMID- 12654811 TI - The stress-responsive dgk gene from Streptococcus mutans encodes a putative undecaprenol kinase activity. AB - We analyzed a previously constructed stress-sensitive Streptococcus mutans mutant Tn-1 strain resulting from disruption by transposon Tn916 of a gene encoding a protein exhibiting amino acid sequence similarity to the Escherichia coli diacylglycerol kinase. It was confirmed that the mutation led to significantly reduced lipid kinase activity, while expression of the intact gene on a plasmid restored both kinase activity and the wild-type phenotype. Further analysis revealed that the product of the dgk gene in S. mutans predominantly recognizes a lipid substrate other than diacylglycerol, most likely undecaprenol, as demonstrated by its efficient phosphorylation and the resistance of the product of the reaction to saponification. The physiological role of the product of the dgk gene as a putative undecaprenol kinase was further supported by a significantly higher sensitivity of the mutant to bacitracin compared with that of the parental strain. PMID- 12654808 TI - P-selectin contributes to severe experimental malaria but is not required for leukocyte adhesion to brain microvasculature. AB - Plasmodium berghei-infected mice, a well-recognized model of experimental cerebral malaria (ECM), exhibit many of the hallmarks of a systemic inflammatory response, with organ damage in brain, lung, and kidneys. Identification of the molecules mediating pathogenesis of the inflammatory response, such as leukocyte adhesion, may lead to new therapies. Indeed, mice lacking the cell adhesion molecule P-selectin were significantly (P = 0.005) protected from death due to P. berghei malaria compared with C57BL/6 controls despite similar parasitemia (P = 0.6) being found in both groups of mice. P-selectin levels assessed by the quantitative dual radiolabeled monoclonal antibody technique increased significantly (P < 0.05) in several organs in C57BL/6 mice infected with P. berghei, supporting the concept of a systemic inflammatory response mediating malarial pathogenesis. Intravital microscopic analysis of the brain microvasculature demonstrated significant (P < 0.001) leukocyte rolling and adhesion in brain venules of P. berghei-infected mice compared with those found in uninfected controls. The maximum leukocyte adhesion occurred on day 6 of P. berghei infection, when the mice become moribund and exhibit marked vascular leakage into the brain, lung, and heart. However, P-selectin levels were significantly (P < 0.005) increased in brain, lung, and kidneys during P. berghei malaria in ECM-resistant BALB/c mice compared with those found in uninfected BALB/c controls, indicating that increased P-selectin alone is not sufficient to mediate malarial pathogenesis. Leukocyte adhesion to brain microvessels of P selectin-deficient mice with P. berghei malaria was similar to that observed in control mice. Collectively, these results indicate that P-selectin is important for the development of malarial pathogenesis but is not required for leukocyte adhesion in brain. PMID- 12654812 TI - Autodisplay: development of an efficacious system for surface display of antigenic determinants in Salmonella vaccine strains. AB - To optimize antigen delivery by Salmonella vaccine strains, a system for surface display of antigenic determinants was established by using the autotransporter secretion pathway of gram-negative bacteria. A modular system for surface display allowed effective targeting of heterologous antigens or fragments thereof to the bacterial surface by the autotransporter domain of AIDA-I, the Escherichia coli adhesin involved in diffuse adherence. A major histocompatibility complex class II-restricted epitope, comprising amino acids 74 to 86 of the Yersinia enterocolitica heat shock protein Hsp60 (Hsp60(74-86)), was fused to the AIDA-I autotransporter domain, and the resulting fusion protein was expressed at high levels on the cell surface of E. coli and Salmonella enterica serovar Typhimurium. Colonization studies in mice vaccinated with Salmonella strains expressing AIDA-I fusion proteins demonstrated high genetic stability of the generated vaccine strain in vivo. Furthermore, a pronounced T-cell response against Yersinia Hsp60(74-86) was induced in mice vaccinated with a Salmonella vaccine strain expressing the Hsp60(74-86)-AIDA-I fusion protein. This was shown by monitoring Yersinia Hsp60-stimulated IFN-gamma secretion and proliferation of splenic T cells isolated from vaccinated mice. These results demonstrate that the surface display of antigenic determinants by the autotransporter pathway deserves special attention regarding the application in live attenuated Salmonella vaccine strains. PMID- 12654814 TI - Signaling through the T1/ST2 molecule is not necessary for Th2 differentiation but is important for the regulation of type 1 responses in nonhealing Leishmania major infection. AB - T1/ST2 is a stable cell surface marker selectively expressed on type 2 T helper (Th2) effector cells. Since nonhealing Leishmania major infections in susceptible BALB/c mice have been ascribed to a polarized Th2 response, we used an anti T1/ST2 monoclonal antibody (MAb) or a T1-Fc fusion protein to investigate the role of CD4+ T1/ST2(+) Th2 cells in experimental leishmaniasis. We show that interfering with T1/ST2 signaling had no effect on lesion development or parasite replication; however, it induced a significantly higher type 1 response and an enhanced capacity of CD4+ T cells to respond to interleukin 12 (IL-12). Surprisingly, even in the presence of an elevated Th1 response, the production of antigen-specific type 2 cytokines was not altered in the group of mice treated with the anti-T1/ST2 MAb or the T1-Fc fusion protein. To characterize further this Th2 response, we assessed the cytokine profile of CD4+ T cells and found that interfering with T1/ST2 signaling did not alter the cytokine profile of CD4+ T1/ST2(+) T cells. These results show that T1/ST2 signaling is not necessary for the differentiation of naive CD4+ T cells into antigen-specific CD4+ T1/ST2(+) Th2 cells. In addition to CD4+ T1/ST2(+) T cells, we detected another subpopulation of CD4+ Th2 cells, negative for the expression of T1/ST2, that could differentiate in vivo in response to L. major infection. Taken together, our results suggest that CD4+ T1/ST2(+) Th2 cells but not CD4+ T1/ST2(-) Th2 cells can downregulate the Th1 response during the course of a nonhealing L. major infection through a mechanism that is independent of IL-4 or IL-10. PMID- 12654813 TI - Synthetic peptides identify promiscuous human Th1 cell epitopes of the secreted mycobacterial antigen MPB70. AB - MPB70 is a secreted protein of Mycobacterium bovis and Mycobacterium tuberculosis which stimulates both cellular and humoral immune responses during infection with bovine and human tubercle bacilli. In addition, vaccination with MPB70 has been shown to induce Th1 cell responses and protection in animal models of tuberculosis. The present study was carried out to map the dominant human Th1 cell epitopes of MPB70 in relation to major histocompatibility complex (MHC) class II restriction in healthy subjects showing strong T-cell responses to complex mycobacterial antigens. Peripheral blood mononuclear cells (PBMC) from HLA-DR-typed donors were tested with complex mycobacterial antigens (whole-cell M. tuberculosis and M. tuberculosis culture filtrates), with MPB70 purified from the culture filtrate of M. bovis BCG Tokyo, and with 13 synthetic peptides (25 mers overlapping by 10 residues) covering the sequence of MPB70. The donors that responded to the complex antigens and MPB70 also responded to the cocktail of synthetic MPB70 peptides. Testing of PBMC with individual peptides showed that peptides p5 (amino acids [aa] 61 to 85), p6 (aa 76 to 100), p8 (aa 106 to 130), p12 (aa 166 to 190), and p13 (aa 181 to 193) were most frequently recognized in proliferation and gamma interferon (IFN-gamma) assays. Testing of antigen specific CD4(+) T-cell lines with the individual peptides of MPB70 confirmed that peptides p8, p12, and p13 contain immunodominant Th1 cell epitopes of MPB70. MHC restriction analysis with HLA-typed donors showed that MPB70 and its immunodominant peptides were presented to T cells promiscuously. The T-cell lines responding to MPB70 and peptides p8, p12, and p13 in IFN-gamma assays mediated antigen-peptide-specific cytotoxic activity against monocytes/macrophages pulsed with the whole-protein antigen or the peptides. In conclusion, the promiscuous recognition of MPB70 and its immunodominant peptide defined epitopes (aa 106 to 130 and 166 to 193) by IFN-gamma-producing Th1 cells supports possible application of this secreted antigen to subunit vaccine design. PMID- 12654815 TI - Mutation of luxS affects biofilm formation in Streptococcus mutans. AB - Quorum sensing is a bacterial mechanism for regulating gene expression in response to changes in population density. Many bacteria are capable of acyl homoserine lactone-based or peptide-based intraspecies quorum sensing and luxS dependent interspecies quorum sensing. While there is good evidence about the involvement of intraspecies quorum sensing in bacterial biofilm, little is known about the role of luxS in biofilm formation. In this study, we report for the first time that luxS-dependent quorum sensing is involved in biofilm formation of Streptococcus mutans. S. mutans is a major cariogenic bacterium in the multispecies bacterial biofilm commonly known as dental plaque. An ortholog of luxS for S. mutans was identified using the data available in the S. mutans genome project (http://www.genome.ou.edu/smutans.html). Using an assay developed for the detection of the LuxS-associated quorum sensing signal autoinducer 2 (AI 2), it was demonstrated that this ortholog was able to complement the luxS negative phenotype of Escherichia coli DH5alpha. It was also shown that AI-2 is indeed produced by S. mutans. AI-2 production is maximal during mid- to late-log growth in batch culture. Mutant strains devoid of the luxS gene were constructed and found to be defective in producing the AI-2 signal. There are also marked phenotypic differences between the wild type and the luxS mutants. Microscopic analysis of in vitro-grown biofilm structure revealed that the luxS mutant biofilms adopted a much more granular appearance, rather than the relatively smooth, confluent layer normally seen in the wild type. These results suggest that LuxS-dependent signal may play an important role in biofilm formation of S. mutans. PMID- 12654816 TI - Recognition of mycobacterial epitopes by T cells across mammalian species and use of a program that predicts human HLA-DR binding peptides to predict bovine epitopes. AB - Bioinformatics tools have the potential to accelerate research into the design of vaccines and diagnostic tests by exploiting genome sequences. The aim of this study was to assess whether in silico analysis could be combined with in vitro screening methods to rapidly identify peptides that are immunogenic during Mycobacterium bovis infection of cattle. In the first instance the M. bovis derived protein ESAT-6 was used as a model antigen to describe peptides containing T-cell epitopes that were frequently recognized across mammalian species, including natural hosts for tuberculosis (humans and cattle) and small animal models of tuberculosis (mice and guinea pigs). Having demonstrated that some peptides could be recognized by T cells from a number of M. bovis-infected hosts, we tested whether a virtual-matrix-based human prediction program (ProPred) could identify peptides that were recognized by T cells from M. bovis infected cattle. In this study, 73% of the experimentally defined peptides from 10 M. bovis antigens that were recognized by bovine T cells contained motifs predicted by ProPred. Finally, in validating this observation, we showed that three of five peptides from the mycobacterial antigen Rv3019c that were predicted to contain HLA-DR-restricted epitopes were recognized by T cells from M. bovis infected cattle. The results obtained in this study support the approach of using bioinformatics to increase the efficiency of epitope screening and selection. PMID- 12654817 TI - Cryptococcus neoformans with a mutation in the tetratricopeptide repeat containing gene, CCN1, causes subcutaneous lesions but fails to cause systemic infection. AB - We studied a Cryptococcus neoformans strain that caused feline chronic nasal granuloma without disseminated disease. This strain, B-4551, grows at temperatures up to 35 degrees C and fails to cause systemic infection in mice. Many cells of B-4551 formed short hyphal elements in feline nasal tissue and occasionally at 35 degrees C in vitro. A complementation and sequence analysis revealed that the temperature-sensitive (Ts) phenotype of B-4551 was due to deletion of a lysine residue in the cryptococcal CCN1 gene. B-4551 complemented with the wild type CCN1 gene grew at 37 degrees C and caused fatal systemic infection in mice. The CCN1 gene encodes a protein containing 16 copies of a tetratricopeptide repeat. CCN1 is homologous to the Saccharomyces cerevisiae CLF1 gene, which is required for pre-mRNA splicing, cell cycle progression, and DNA replication, and to the Drosophila melanogaster crn gene, which is involved in neurogenesis. CLF1 complemented the Ts phenotype of B-4551. CCN1, however, failed to rescue the clf1 mutant in S. cerevisiae. These results indicate that the Ccn1p may not be as functionally diverse as Clf1p in yeast. PMID- 12654818 TI - Anaplasma phagocytophilum reduces neutrophil apoptosis in vivo. AB - Ovine neutrophils spontaneously underwent apoptosis during culture in vitro, as assessed by morphological changes and exposure of annexin V binding sites on their cell surfaces. The addition of conditioned medium from concanavalin A treated ovine peripheral blood mononuclear cells (PBMC) could partially protect against this progression into apoptosis, but dexamethasone and sodium butyrate could not. Actinomycin D accelerated the rate at which ovine neutrophils underwent apoptosis. Neutrophils isolated from sheep experimentally infected with Anaplasma phagocytophilum showed significantly delayed apoptosis during culture ex vivo, and the addition of conditioned medium from PBMC to these cells could not delay apoptosis above the protective effects observed after in vivo infection. The ability of neutrophils from A. phagocytophilum-infected sheep to activate a respiratory burst was increased compared to the activity measured in neutrophils from uninfected sheep, but chemotaxis was decreased in neutrophils from infected sheep. These data are the first demonstration that in vivo infection with A. phagocytophilum results in changes in rates of apoptosis of infected immune cells. This may help explain how these bacteria replicate in these normally short-lived cells. PMID- 12654820 TI - Effects of zinc deficiency and pneumococcal surface protein a immunization on zinc status and the risk of severe infection in mice. AB - Streptococcus pneumoniae is a major cause of illness and death in children in developing countries. In these children, zinc deficiency is associated with an increased risk of acute respiratory tract infections, which can be reduced by daily zinc administration. Severe infections decrease zinc levels in plasma and may thereby move individuals with preexisting low zinc stores into a vicious cycle of infection and unavailable zinc. Pneumococcal surface protein A (PspA) has emerged as a promising vaccine candidate, and immunization with this antigen protects animals from pneumococcal infection. In an animal experiment, we measured the effect of zinc depletion on the immune response to parenterally administrated PspA and assessed the effect of this PspA vaccination and zinc depletion on the severity of pneumococcal infection and on zinc status. Mice were kept on different diets for 5 weeks, immunized twice 14 days apart, and challenged intranasally with S. pneumoniae. Mice on the zinc-deficient diet showed substantially reduced immune responses to PspA, more extensive pneumococcal colonization in the nasal mucosa, more severe infections, and an increased risk of death. PspA immunization reduced the risk of severe disease, and the reduction in severity was reflected in substantially reduced zinc depletion from bones. PMID- 12654819 TI - Resistance to acute babesiosis is associated with interleukin-12- and gamma interferon-mediated responses and requires macrophages and natural killer cells. AB - We examined the role of the cytokines gamma interferon (IFN-gamma) and interleukin-12 (IL-12) in the model of acute babesiosis with the WA1 Babesia. Mice genetically deficient in IFN-gamma-mediated responses (IFNGR2KO mice) and IL 12-mediated responses (Stat4KO mice) were infected with the WA1 Babesia, and observations were made on the course of infection and cytokine responses. Levels of IFN-gamma and IL-12 in serum increased 24 h after parasite inoculation. The augmented susceptibility observed in IFNGR2KO and Stat-4KO mice suggests that the early IL-12- and IFN-gamma-mediated responses are involved in protection against acute babesiosis. Resistance appears to correlate with an increase in nitric oxide (NO) production. In order to assess the contribution of different cell subsets to resistance against the parasite, we also studied mice lacking B cells, CD4+ T cells, NK cells, and macrophages. Mice genetically deficient in B lymphocytes or CD4+ T lymphocytes were able to mount protective responses comparable to those of immunosufficient mice. In contrast, in vivo depletion of macrophages or NK cells resulted in elevated susceptibility to the infection. Our observations suggest that a crucial part of the response that protects from the pathogenic Babesia WA1 is mediated by macrophages and NK cells, probably through early production of IL-12 and IFN-gamma, and induction of macrophage-derived effector molecules like NO. PMID- 12654821 TI - Pathogenic Yersinia enterocolitica strains increase the outer membrane permeability in response to environmental stimuli by modulating lipopolysaccharide fluidity and lipid A structure. AB - Pathogenic biotypes of Yersinia enterocolitica (serotypes O:3, O:8, O:9, and O:13), but not environmental biotypes (serotypes O:5, O:6, O:7,8, and O:7,8,13,19), increased their permeability to hydrophobic probes when they were grown at pH 5.5 or in EGTA-supplemented (Ca(2+)-restricted) media at 37 degrees C. A similar observation was also made when representative strains of serotypes O:8 and O:5 were tested after brief contact with human monocytes. The increase in permeability was independent of the virulence plasmid. The role of lipopolysaccharide (LPS) in this phenomenon was examined by using Y. enterocolitica serotype O:8. LPS aggregates of bacteria grown in acidic or EGTA supplemented broth took up more N-phenylnaphthylamine than LPS aggregates of bacteria grown in standard broth and also showed a marked increase in acyl chain fluidity which correlated with permeability, as determined by measurements obtained in the presence of hydrophobic dyes. No significant changes in O-antigen polymerization were observed, but lipid A acylation changed depending on the growth conditions. In standard medium at 37 degrees C, there were hexa-, penta-, and tetraacyl lipid A forms, and the pentaacyl form was dominant. The amount of tetraacyl lipid A increased in EGTA-supplemented and acidic media, and hexaacyl lipid A almost disappeared under the latter conditions. Our results suggest that pathogenic Y. enterocolitica strains modulate lipid A acylation coordinately with expression of virulence proteins, thus reducing LPS packing and increasing outer membrane permeability. The changes in permeability, LPS acyl chain fluidity, and lipid A acylation in pathogenic Y. enterocolitica strains approximate the characteristics in Yersinia pseudotuberculosis and Yersinia pestis and suggest that there is a common outer membrane pattern associated with pathogenicity. PMID- 12654822 TI - Helicobacter pylori mutants defective in RuvC Holliday junction resolvase display reduced macrophage survival and spontaneous clearance from the murine gastric mucosa. AB - Homologous recombination contributes to the extraordinary genetic diversity of Helicobacter pylori and may be critical for surface antigen expression and adaptation to environmental challenges within the stomach. We generated isogenic, nonpolar H. pylori ruvC mutants to investigate the function of RuvC, a Holliday junction endonuclease that resolves recombinant joints into nicked duplex products. Inactivation of ruvC reduced the frequency of homologous recombination of H. pylori between 17- and 45-fold and increased sensitivity to DNA-damaging agents and the antimicrobial agents levofloxacin and metronidazole. The H. pylori ruvC mutants were more susceptible to oxidative stress and exhibited reduced survival within macrophages. Experiments with the H. pylori SS1 mouse model revealed that the 50% infective dose of the ruvC mutant was approximately 100 fold higher than that of the wild-type SS1 strain. Although the ruvC mutant was able to establish colonization with bacterial loads that were initially similar to those of the parental SS1 strain, infection was spontaneously cleared from the murine gastric mucosa over periods that varied from 36 to 67 days. These results demonstrate that, in this infection model, RuvC is essential for continued survival of H. pylori in vivo and raises the possibility that inactivation of ruvC might be of value in an attenuated vaccine strain. PMID- 12654824 TI - Different genetic control of cutaneous and visceral disease after Leishmania major infection in mice. AB - The mouse strains BALB/cHeA (BALB/c) and STS/A (STS) are susceptible and resistant to Leishmania major-induced disease, respectively. We analyzed this difference using recombinant congenic (RC) BALB/c-c-STS/Dem (CcS/Dem) strains that carry different random subsets of 12.5% genes of the strain STS in a BALB/c background. Previously, testing the resistant strain CcS-5, we found five novel Lmr (Leishmania major response) loci, each associated with a different combination of pathological and immunological reactions. Here we analyze the response of RC strain CcS-16, which is even more susceptible to L. major than BALB/c. In the (CcS-16 x BALB/c)F(2) hybrids we mapped three novel loci that influence cutaneous or visceral pathology. Lmr14 (chromosome 2) controls splenomegaly and hepatomegaly. On the other hand Lmr15 (chromosome 11) determines hepatomegaly only, and Lmr13 (chromosome 18) determines skin lesions only. These data confirm the complex control of L. major-induced pathology, where cutaneous and visceral pathology are controlled by different combinations of genes. It indicates organ-specific control of antiparasite responses. The definition of genes controlling these responses will permit a better understanding of pathways and genetic diversity underlying the different disease phenotypes. PMID- 12654823 TI - Induction of mosquitocidal activity in mice immunized with Anopheles gambiae midgut cDNA. AB - Vaccines that induce mosquito-killing (mosquitocidal) activity could substantially reduce the transmission of certain mosquito-borne diseases, especially vaccines against African malaria vectors, such as the mosquito Anopheles gambiae. To generate and characterize antimosquito immunity we immunized groups of mice with two individual A. gambiae midgut cDNAs, Ag-Aper1 (a secreted peritrophic matrix protein) and AgMuc1 (a midgut-bound mucin), and an A. gambiae midgut cDNA library from blood-fed mosquitoes. We observed significantly increased mortality among mosquitoes that fed on either the AgMuc1- or the cDNA library-immunized mice compared to that of controls, but no differences were observed among those fed on Ag-Aper1-immunized mice. Analysis of the humoral and cellular immune responses from mice showed that the induced mosquitocidal effect was associated with immune profiles characterized by elevated tumor necrosis factor alpha and gamma interferon cytokine levels and very low antibody titers. Furthermore, an additional immunization of cDNA library-immunized mice with midgut protein shifted immunity toward a Th2-type immune response, characterized by elevated antibody titers and high interleukin-5 and interleukin-10 cytokine levels; importantly, mosquitoes feeding on these mice exhibited no undue mortality. Finally, when immune sera was ingested by mosquitoes through a membrane feeder, no effect on mosquito mortality was observed, indicating that serum factors alone were not responsible for the mosquitocidal effect. Our results demonstrate that mosquitocidal immunity in mice can be consistently generated by midgut cDNA immunization and suggest this cDNA-induced mosquitocidal immunity is cell mediated. PMID- 12654825 TI - Soluble factors released by Toxoplasma gondii-infected astrocytes down-modulate nitric oxide production by gamma interferon-activated microglia and prevent neuronal degeneration. AB - The maintenance of a benign chronic Toxoplasma gondii infection is mainly dependent on the persistent presence of gamma interferon (IFN-gamma) in the central nervous system (CNS). However, IFN-gamma-activated microglia are paradoxically involved in parasitism control and in tissue damage during a broad range of CNS pathologies. In this way, nitric oxide (NO), the main toxic metabolite produced by IFN-gamma-activated microglia, may cause neuronal injury during T. gondii infection. Despite the potential NO toxicity, neurodegeneration is not a common finding during chronic T. gondii infection. In this work, we describe a significant down-modulation of NO production by IFN-gamma-activated microglia in the presence of conditioned medium of T. gondii-infected astrocytes (CMi). The inhibition of NO production was paralleled with recovery of neurite outgrowth when neurons were cocultured with IFN-gamma-activated microglia in the presence of CMi. Moreover, the modulation of NO secretion and the neuroprotective effect were shown to be dependent on prostaglandin E(2) (PGE(2)) production by T. gondii-infected astrocytes and autocrine secretion of interleukin-10 (IL-10) by microglia. These events were partially eliminated when infected astrocytes were treated with aspirin and cocultures were treated with anti-IL-10 neutralizing antibodies and RP-8-Br cyclic AMP (cAMP), a protein kinase A inhibitor. Further, the modulatory effects of CMi were mimicked by the presence of exogenous PGE(2) and by forskolin, an adenylate cyclase activator. Altogether, these data point to a T. gondii-triggered regulatory mechanism involving PGE(2) secretion by astrocytes and cAMP-dependent IL-10 secretion by microglia. This may reduce host tissue inflammation, thus avoiding neuron damage during an established Th1 protective immune response. PMID- 12654827 TI - Exposure of immunocompetent adult mice to Pneumocystis carinii f. sp. muris by cohousing: growth of P. carinii f. sp. muris and host immune response. AB - There has been emerging evidence that immunocompetent hosts can harbor Pneumocystis in their lungs. The purpose of this study was to determine the kinetics of Pneumocystis carinii f. sp. muris infection in adult immunocompetent mice and the host immune response to the organisms. To accomplish this, we exposed adult immunocompetent mice to SCID mice infected with P. carinii f. sp. muris by cohousing. We found that P. carinii f. sp. muris was detectable in the lungs of cohoused immunocompetent mice by PCR by 3 weeks after the beginning of cohousing. At about 4 weeks of cohousing, P. carinii f. sp. muris was readily detectable in the lungs of mice by microscopic techniques. Also at this time, P. carinii f. sp. muris-specific immunoglobulin G was found in the sera of the mice, and CD62(low) CD4- and CD8-positve T cells accumulated in the lungs. Shortly after this immune response, the P. carinii f. sp. muris organisms were cleared from the lungs. Adult mice cohoused for only 1 week also contained P. carinii f. sp. muris cysts detectable by silver staining at 5 and 6 weeks after the beginning of cohousing. We also found that the P. carinii f. sp. muris organisms grew to greater numbers in the lungs of BALB/c mice than in those of C57BL6 mice. This indicates that immunocompetent hosts develop a mild infection with P. carinii f. sp. muris which resolves in 5 to 6 weeks when there is a detectable immune response to the organism. Once an acquired immune response was initiated, the P. carinii f. sp. muris organisms were quickly eliminated without clinical signs of disease. PMID- 12654826 TI - Recombinant gamma interferon stimulates signal transduction and gene expression in alveolar macrophages in vitro and in tuberculosis patients. AB - Tuberculosis is the seventh leading cause of morbidity and mortality in the world, with eight million cases per year. Animal and human studies demonstrate an enrichment of CD4 cells at sites of disease, with a more favorable clinical course when there is a Th1 response with the presence of gamma interferon (IFN gamma). We previously treated patients who had multidrug-resistant tuberculosis with recombinant IFN-gamma (rIFN-gamma) in aerosol form and were able to convert smear-positive cases to smear negative with 12 treatments over 1 month. We hypothesized that rIFN-gamma would induce signal transducer and activator of transcription (STAT) and interferon regulatory factor (IRF) binding activity in alveolar macrophages (AM). AM treated in vitro showed clear upregulation of STAT 1 and IRF-1 by rIFN-gamma. STAT-1 was not activated and IRF-1 was only weakly induced after 1 day of infection by Mycobacterium tuberculosis TN913. In bronchoalveolar lavage (BAL) cells obtained from 10 of 10 tuberculosis patients 10 +/- 2 days post-antituberculosis treatment, there was no detectable STAT-1 or IRF-1 DNA-binding activity. After 4 weeks of treatment with rIFN-gamma aerosol in addition to the antituberculosis drugs, 10 of 10 patients had increased STAT-1, IRF-1, and/or IRF-9 DNA-binding activity in BAL cells from lung segments shown radiographically to be involved and in those shown to be uninvolved. Symptoms and chest radiographs improved, and amounts of macrophage inflammatory cytokines and human immunodeficiency virus type 1 (HIV-1) viral loads (in five of five HIV-1 coinfected patients) declined in the second BAL specimens. rIFN-gamma aerosol induces signal transduction and gene expression in BAL cells and should be evaluated for efficacy in a randomized, controlled clinical trial. PMID- 12654828 TI - Transmission of Anaplasma phagocytophilum to Ixodes ricinus ticks from sheep in the acute and post-acute phases of infection. AB - A total of 60 sheep were exposed to Anaplasma phagocytophilum infection on an enclosed area of Ixodes ricinus-infested pasture in North Wales, United Kingdom, and rapidly acquired acute A. phagocytophilum infections detectable by PCR and blood smear examination. Of the ticks that had engorged in the previous instar on infected sheep, 52% of adult ticks and 28% of nymphs were PCR positive; a significant, 10-fold increase in prevalence compared to that of ticks that engorged on sheep preinfection was observed (P = 0.015). The likelihood that ticks were PCR positive, after feeding on the sheep and molting to the next instar, increased marginally with increasing numbers of infected neutrophils per milliliter of blood of their sheep host (P = 0.068) and increased significantly when they were collected from sheep carrying higher numbers of adult female ticks (P = 0.017), but increasing numbers of feeding nymphs had a significant negative effect on transmission (P = 0.049). The numbers of circulating neutrophils and of infected neutrophils also varied significantly with the numbers of ticks feeding on the sheep when the blood was collected. Our study suggests that ruminants are efficient reservoirs of A. phagocytophilum during the acute and post-acute phases of infection. The risk of ruminant-derived infections may, however, be strongly affected by variations in tick densities, which may influence transmission from acutely infected animals via effects on the numbers of infected cells in the blood and possibly by within-skin modulation of infection. PMID- 12654829 TI - Major histocompatibility complex heterozygote superiority during coinfection. AB - Genes of the major histocompatibility complex (MHC) play a critical role in immune recognition, and many alleles confer susceptibility to infectious and autoimmune diseases. How these deleterious alleles persist in populations is controversial. One hypothesis postulates that MHC heterozygote superiority emerges over multiple infections because MHC-mediated resistance is generally dominant and many allele-specific susceptibilities to pathogens will be masked by the resistant allele in heterozygotes. We tested this hypothesis by using experimental coinfections with Salmonella enterica (serovar Typhimurium C5TS) and Theiler's murine encephalomyelitis virus (TMEV) in MHC-congenic mouse strains where one haplotype was resistant to Salmonella and the other was resistant to TMEV. MHC heterozygotes were superior to both homozygotes in 7 out of 8 comparisons (P = 0.0024), and the mean standardized pathogen load of heterozygotes was reduced by 41% over that of homozygotes (P = 0.01). In contrast, no heterozygote superiority was observed when the MHC haplotype combinations had similar susceptibility profiles to the two pathogens. This is the first experimental evidence for MHC heterozygote superiority against multiple pathogens, a mechanism that would contribute to the evolution of MHC diversity and explain the persistence of alleles conferring susceptibility to disease. PMID- 12654830 TI - Pneumolysin-dependent and -independent gene expression identified by cDNA microarray analysis of THP-1 human mononuclear cells stimulated by Streptococcus pneumoniae. AB - Pneumolysin is an important virulence factor of Streptococcus pneumoniae, interacting with the membranes of host cells to elicit a multitude of inflammatory responses. We used cDNA microarrays to identify genes which are responsive to S. pneumoniae in a pneumolysin-dependent and -independent fashion. The THP-1 human monocytic cell line was coincubated for 3 h with medium alone, with the virulent type 2 S. pneumoniae strain D39, or with the isogenic strain PLN, which does not express pneumolysin. RNA was isolated from the monocytes and hybridized on cDNA microarrays. Of 4,133 genes evaluated, 142 were found to be responsive in a pneumolysin-dependent fashion, whereas 40 were found to be responsive independent of pneumolysin. Genes that were up-regulated in cells exposed to D39 relative to those exposed to PLN included genes encoding proteins such as mannose binding lectin 1, lysozyme, alpha-1 catenin, cadherin 17, caspases 4 and 6, macrophage inflammatory protein 1beta (MIP-1beta), interleukin 8 (IL-8), monocyte chemotactic protein 3 (MCP-3), IL-2 receptor beta (IL-2Rbeta), IL-15 receptor alpha (IL-15Ralpha), interferon receptor 2, and prostaglandin E synthase. Down-regulated genes included those encoding complement component receptor 2/CD21, platelet-activating factor acetylhydrolase, and oxidized low density lipoprotein receptor 1 (OLR1). Pneumolysin-independent responses included down-regulation of the genes encoding CD68, CD53, CD24, transforming growth factor beta2, and signal transducers and activators of transcription 1. These results demonstrate the striking effects of pneumolysin on the host cell upon exposure to S. pneumoniae. PMID- 12654831 TI - Leishmania donovani-induced expression of suppressor of cytokine signaling 3 in human macrophages: a novel mechanism for intracellular parasite suppression of activation. AB - Leishmania donovani protozoan parasites, the causative agent of visceral leishmaniasis, establish an infection partly by interfering with cytokine signaling in the host macrophages. Therefore, we investigated the expression of the suppressor of cytokine signaling (SOCS) genes in human macrophages infected with L. donovani. The expression of SOCS3 mRNA was induced transiently after exposure to live or heat-killed parasites, but not purified lipophosphoglycan, while that of other SOCS genes remained unchanged. SOCS3 gene expression was not dependent on phagocytosis or on cytokines released by L. donovani-infected macrophages, such as interleukin-1beta or tumor necrosis factor alpha. In addition, Leishmania used a different signaling pathway(s) than bacterial lipopolysaccharide to induce SOCS3 mRNA, as indicated by the kinetics of induction and sensitivity to polymyxin B inhibition. Finally, phosphorylation of the STAT1 transcription factor was significantly reduced in L. donovani-infected macrophages and required de novo transcription. The induction of SOCS3 provides a potent inhibitory mechanism by which intracellular microorganisms may suppress macrophage activation and interfere with the host immune response. PMID- 12654832 TI - Salmonella infection induces a hypersecretory phenotype in human intestinal xenografts by inducing cyclooxygenase 2. AB - Enteric Salmonella infection is accompanied by inflammation and diarrhea, and yet little is known about its effects on intestinal epithelial physiology. Since species differences limit the utility of animal tissues and cell lines lack relevant cell-cell interactions, we have used a human model of fetal intestine grown as xenografts in SCID mice. We investigated here the effects of Salmonella enterica serovar Typhimurium SL1344 on xenograft ion transport. Harvested xenografts were stripped of seromuscular layers by blunt dissection, infected with Salmonella, and mounted in Ussing chambers. Salmonella infection for 1 h increased baseline ion transport without altering tissue conductance or morphology. The increased transport was blocked by the cyclooxygenase inhibitor, indomethacin, or the specific Cox-2 inhibitor, NS-398. Further, xenografts infected for 2 h showed increased secretory responses to the calcium-dependent agonist, carbachol, and the cyclic AMP-dependent agonists prostaglandin E(2) (PGE(2)) and forskolin, which were blocked by indomethacin. Western blot experiments revealed that infection was accompanied by increased cyclooxygenase 2 (Cox-2) expression, with no change in Cox-1 levels. Immunoassay demonstrated basolateral PGE(2) release, which was inhibited by indomethacin. Histological examination of infected xenografts illustrated that upregulated Cox-2 expression was restricted to the epithelium and that little or no invasion of the tissue by Salmonella occurred for up to 2 h. In summary, Salmonella infection rapidly increases Cox-2 expression in human intestinal tissue, accounting for increased epithelial ion transport characteristic of infectious diarrhea. PMID- 12654833 TI - Interactions between Brucella melitensis and human phagocytes: bacterial surface O-Polysaccharide inhibits phagocytosis, bacterial killing, and subsequent host cell apoptosis. AB - Brucellae are gram-negative intracellular pathogens that survive and multiply within host phagocytic cells. Smooth organisms present O-polysaccharides (OPS) on their surface. The wboA gene, which codes for the enzyme glycosyl transferase, is essential for the assembly of O-chain in Brucella. Deletion of wboA in smooth, virulent B. melitensis 16M results in a rough mutant designated WRR51. Unlike B. abortus, both smooth and rough strains of B. melitensis are resistant to complement-mediated killing. To determine the role of surface OPS in the interactions of B. melitensis with monocytes/macrophages (M/M), 16M and WRR51 were transformed with the plasmid pBBR1MCS-6y encoding green fluorescent protein, and the transformants were used to infect human mononuclear phagocytes with and without fresh human serum as a source of complement. Human monocytes were cultured in the presence of macrophage colony-stimulating factor to allow their differentiation into macrophages during the course of infection. Intracellular bacteria were easily visualized using fluorescence microscopy. Infection in M/M, identified by surface staining and fate of infected phagocytes, was quantitated by flow cytometry. Rough bacteria were internalized, with no requirement for opsonization by serum, at a higher rate than smooth organisms. Smooth B. melitensis survived and multiplied for at least 6 days inside M/M, but rough organisms were eliminated by death of the infected cells. In human monocytes cultured for 1 day without serum in order to trigger the apoptotic pathway, infection by rough brucellae accelerated phagocyte death; smooth brucellae inhibited apoptosis. This study suggests that the presence of surface OPS on live B. melitensis benefits the bacterium by preventing the death of macrophages, Brucella's preferred target for intracellular replication. PMID- 12654834 TI - Flagellin of enteropathogenic Escherichia coli stimulates interleukin-8 production in T84 cells. AB - The type III secretion system (TTSS) of enteropathogenic Escherichia coli (EPEC) has been associated with the ability of these bacteria to induce secretion of proinflammatory cytokines, including interleukin-8 (IL-8), in cultured epithelial cells. However, the identity of the effector molecule directly involved in this event is unknown. In this study, we determined that the native flagellar filament and its flagellin monomer are activators of IL-8 release in T84 epithelial cells. Supernatants of wild-type EPEC strain E2348/69 and its isogenic mutants deficient in TTSS (escN) and in production of intimin (eae), grown in Luria-Bertani broth, elicited similar amounts of IL-8 secretion by T84 cells. In contrast, supernatants of EPEC fliC mutants and of B171, a nonflagellated EPEC strain, were defective in inducing IL-8 release, a phenotype that was largely restored by complementation of the fliC gene in the mutant lacking flagella. Purified flagella from E. coli K-12, EPEC serotypes H6 and H34, and enterohemorrhagic E. coli serotype H7 all induced IL-8 release in T84 cells. Induction of IL-8 by purified flagella or His-tagged FliC from EPEC strain E2348/69 was dose dependent and was blocked by a polyclonal anti-H6 antibody. Finally, the mitogen-activated protein kinases (Erk1 and -2 and Jnk) were phosphorylated in flagellin-treated T84 cells, and inhibition of the p38 and Erk pathways significantly decreased the IL-8 response induced by EPEC flagellin. Our data clearly indicate that FliC of EPEC is sufficient to induce IL-8 release in T84 cells and that activation of the Erk and p38 pathways is required for IL-8 induction. PMID- 12654835 TI - CesD2 of enteropathogenic Escherichia coli is a second chaperone for the type III secretion translocator protein EspD. AB - Enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli are extracellular pathogens that employ a type III secretion system to export translocator and effector proteins, proteins which facilitates colonization of the mucosal surface of the intestine via formation of attaching and effacing (A/E) lesions. The genes encoding the proteins for A/E lesion formation are located on a pathogenicity island, termed the locus of enterocyte effacement (LEE), which contains eae encoding intimin as well as the type III secretion system and effector genes. Many type III secreted proteins are stabilized and maintained in a secretion-competent conformation in the bacterial cytosol by specific chaperone proteins. Three type III chaperones have been described thus far within the EPEC LEE region: CesD, for the translocator proteins EspB and EspD; CesT, for the effector proteins Tir and Map; and CesF, for EspF. In this study we report the characterization of CesD2 (previously Orf27), a second LEE encoded chaperone for EspD. We show specific CesD2-EspD protein interaction which appears to be necessary for proper EspD secretion in vitro and pathogenesis in vivo as demonstrated in the A/E-lesion-forming mouse pathogen Citrobacter rodentium. PMID- 12654836 TI - An Escherichia coli MG1655 lipopolysaccharide deep-rough core mutant grows and survives in mouse cecal mucus but fails to colonize the mouse large intestine. AB - The ability of E. coli strains to colonize the mouse large intestine has been correlated with their ability to grow in cecal and colonic mucus. In the present study, an E. coli MG1655 strain was mutagenized with a mini-Tn5 Km (kanamycin) transposon, and mutants were tested for the ability to grow on agar plates with mouse cecal mucus as the sole source of carbon and nitrogen. One mutant, designated MD42 (for mucus defective), grew poorly on cecal-mucus agar plates but grew well on Luria agar plates and on glucose minimal-agar plates. Sequencing revealed that the insertion in MD42 was in the waaQ gene, which is involved in lipopolysaccharide (LPS) core biosynthesis. Like "deep-rough" E. coli mutants, MD42 was hypersensitive to sodium dodecyl sulfate (SDS), bile salts, and the hydrophobic antibiotic novobiocin. Furthermore, its LPS core oligosaccharide was truncated, like that of a deep-rough mutant. MD42 initially grew in the large intestines of streptomycin-treated mice but then failed to colonize (<10(2) CFU per g of feces), whereas its parent colonized at levels between 10(7) and 10(8) CFU per g of feces. When mouse cecal mucosal sections were hybridized with an E. coli-specific rRNA probe, MD42 was observed in cecal mucus as clumps 24 h postfeeding, whereas its parent was present almost exclusively as single cells, suggesting that clumping may play a role in preventing MD42 colonization. Surprisingly, MD42 grew nearly as well as its parent during growth in undiluted, highly viscous cecal mucus isolated directly from the mouse cecum and, like its parent, survived well after reaching stationary phase, suggesting that there are no antimicrobials in mucus that prevent MD42 colonization. After mini-mariner transposon mutagenesis, an SDS-resistant suppressor mutant of MD42 was isolated. The mini-mariner insertion was shown to be in the bipA gene, a known regulator of E. coli surface components. When grown in Luria broth, the LPS core of the suppressor mutant remained truncated; however, the LPS core was not truncated when the suppressor mutant was grown in the presence of SDS. Moreover, when the suppressor mutant was grown in the presence of SDS and fed to mice, it colonized the mouse large intestine. Collectively, the data presented here suggest that BipA may play a role in E. coli MG1655 LPS core biosynthesis and that because MD42 forms clumps in intestinal mucus, it is unable to colonize the mouse large intestine. PMID- 12654838 TI - Mapping the binding domain of the F18 fimbrial adhesin. AB - F18 fimbrial Esherichia coli strains are associated with porcine postweaning diarrhea and pig edema disease. Recently, the FedF subunit was identified as the adhesin of the F18 fimbriae. In this study, adhesion domains of FedF were further studied by constructing deletions within the fedF gene and expressing FedF proteins with deletions either together with the other F18 fimbrial subunits or as fusion proteins tagged with maltose binding protein. The region essential for adhesion to porcine intestinal epithelial cells was mapped between amino acid residues 60 and 109 of FedF. To map the binding domain even more closely, all eight charged amino acid residues within this region were independently replaced by alanine. Three of these single point mutants expressing F18 fimbriae exhibited significantly diminished capabilities to adhere to porcine epithelial cells in vitro. In addition, a triple point mutation and a double point mutation completely abolished receptor adhesiveness. The result further confirmed that the region between amino acid residues 60 and 109 is essential for the binding of F18 fimbriae to their receptor. In addition, the adhesion capability of the binding domain was eliminated after treatment with iodoacetamide, suggesting the formation of a disulfide bridge between Cys-63 and Cys-83, whereas Cys-111 and Cys-116 could be deleted without affecting the binding ability of FedF. PMID- 12654837 TI - Involvement of myeloid dendritic cells in the development of gastric secondary lymphoid follicles in Helicobacter pylori-infected neonatally thymectomized BALB/c mice. AB - We previously described an animal model of Helicobacter pylori-induced follicular gastritis in neonatally thymectomized (nTx) mice. However, it is still not clear whether antigen-presenting dendritic cells (DCs) in the stomach have a role in the development of secondary follicles in H. pylori-infected nTx mice. We investigated the distribution of DC subsets using this model and examined their roles. To identify lymphoid and myeloid DCs, sections were stained with anti CD11c (pan-DC marker) in combination with anti-CD8alpha (lymphoid DC marker) or anti-CD11b (myeloid DC marker) and were examined with a confocal microscope. Expression of macrophage inflammatory protein 3alpha (MIP-3alpha), which chemoattracts immature DCs, was analyzed by real-time PCR and immunohistochemistry. Follicular dendritic cells (FDCs) were stained with anti SKY28 antibodies. In noninfected nTx mice, a few myeloid and lymphoid DCs were observed in the bottom portion of the lamina propria, whereas in H. pylori infected nTx mice, there was an increased influx of myeloid DCs throughout the lamina propria. FDC staining was also observed in the stomachs of members of the infected group. MIP-3alpha gene expression was upregulated in the infected nTx group, and the immunohistochemistry analysis revealed MIP-3alpha-positive epithelial cells. These data suggest that H. pylori infection upregulates MIP 3alpha gene expression in gastric epithelial cells and induces an influx of myeloid DCs in the lamina propria of the gastric mucosa in nTx mice. Myeloid DCs and FDCs might contribute to the development of gastric secondary lymphoid follicles in H. pylori-infected nTx mice. PMID- 12654839 TI - Identification and characterization of a conserved, stage-specific gene product of Plasmodium falciparum recognized by parasite growth inhibitory antibodies. AB - We have identified a novel conserved protein of Plasmodium falciparum, designated D13, that is stage-specifically expressed in asexual blood stages of the parasite. The predicted open reading frame (ORF) D13 contains 863 amino acids with a calculated molecular mass of 99.7 kDa and displays a repeat region composed of pentapeptide motives. Northern blot analysis with lysates of synchronized blood stage parasites showed that D13 is highly expressed at the mRNA level during schizogony. The first N'-terminal 138 amino acids of D13 were expressed in Escherichia coli and the purified protein was used to generate anti D13 monoclonal antibodies (MAbs). Using total lysates of blood stage parasites and Western blot analysis, these MAbs stained one single band of approximately 100 kDa, corresponding to the predicted molecular mass of ORF D13. Western blot analysis demonstrated further that D13 is expressed during schizogony, declines rapidly in early ring stages and is undetectable in trophozoites. D13 protein is localized in individual merozoites in a distinct area, as demonstrated by indirect immunofluorescence analysis. After subcellular fractionation, D13 was confined to the pelleted fraction of the parasite lysate and its extraction by alkaline carbonate buffer treatment indicated that D13 is not a membrane-integral protein. Inclusion of certain anti-D13 MAbs into in vitro cultures of blood stage parasites resulted in considerable reduction in parasite growth. The N'-terminal domain encompassing 158 amino acids is 94 and 95%, respectively, identical at the amino acid level between Plasmodium knowlesi, Plasmodium yoelii, and P. falciparum. In summary, we describe a novel stage-specifically expressed, highly conserved gene product of P. falciparum that is recognized by parasite growth inhibitory antibodies. PMID- 12654840 TI - Rapid protection of gnotobiotic pigs against experimental salmonellosis following induction of polymorphonuclear leukocytes by avirulent Salmonella enterica. AB - Oral inoculation of 5-day-old gnotobiotic pigs with Salmonella enterica serovar Typhimurium strain F98 resulted in severe enteritis and invasive disease. Preinoculation 24 h earlier with an avirulent mutant of Salmonella enterica serovar Infantis (1326/28) completely prevented disease for up to 14 days (when the experiment was terminated). S. enterica serovar Infantis colonized the alimentary tract well, with high bacterial counts in the intestinal lumen but with almost no invasion into the tissues. Unprotected pigs had high S. enterica serovar Typhimurium counts in the intestines, blood, and major nonintestinal organs. Recovery of this strain from the blood and major organs in S. enterica serovar Infantis-protected pigs was substantially reduced despite the fact that intestinal counts were also very high. Protection against disease thus did not involve a colonization exclusion phenomenon. Significant (P < 0.05) infiltration of monocytes/macrophages was observed in the submucosal regions of the intestines of both S. enterica serovar Infantis-protected S. enterica serovar Typhimurium challenged pigs and unprotected S. enterica serovar Typhimurium-challenged pigs. However, only polymorphonuclear neutrophils (PMNs) were observed throughout the villus, where significant (P < 0.05) numbers infiltrated the lamina propria and the subnuclear and supranuclear regions of the epithelia, indicating that PMN induction and positioning following S. enterica serovar Infantis inoculation was consistent with rapid protection against the challenge strain. Similarly, in vitro experiments using a human fetal intestinal epithelial cell line (INT 407) demonstrated that, although significantly (P < 0.05) fewer S. enterica serovar Infantis than S. enterica serovar Typhimurium organisms invaded the monolayers, S. enterica serovar Infantis induced an NF-kappaB response and significantly (P < 0.05) raised interleukin 8 levels and transmigration of porcine PMN. The results of this study suggest that attenuated Salmonella strains can protect the immature intestine against clinical salmonellosis by PMN induction. They also demonstrate that PMN induction is not necessarily associated with clinical symptoms and/or intestinal pathology. PMID- 12654841 TI - Pulmonary necrosis resulting from DNA vaccination against tuberculosis. AB - The use of DNA constructs encoding mycobacterial proteins is a promising new approach to vaccination against tuberculosis. A DNA vaccine encoding the hsp60 molecule of Mycobacterium leprae has previously been shown to protect against intravenous infection of mice with Mycobacterium tuberculosis in both the prophylactic and immunotherapeutic modes. It is shown here, however, that this vaccine was not effective in a more realistic aerosol infection model or in a model of latent tuberculosis in the lungs. Moreover, when given in an immunotherapeutic model the immunized mice developed classical Koch reactions characterized by multifocal discrete regions of cellular necrosis throughout the lung granulomas. Similar and equally severe reactions were seen in mice given a vaccine with DNA coding for the Ag85 antigen of M. tuberculosis. This previously unanticipated safety problem indicates that DNA vaccines should be used with caution in individuals who may have already been exposed to tuberculosis. PMID- 12654842 TI - Group A Streptococcus gene expression in humans and cynomolgus macaques with acute pharyngitis. AB - The molecular mechanisms used by group A Streptococcus (GAS) to survive on the host mucosal surface and cause acute pharyngitis are poorly understood. To provide new information about GAS host-pathogen interactions, we used real-time reverse transcription-PCR (RT-PCR) to analyze transcripts of 17 GAS genes in throat swab specimens taken from 18 pediatric patients with pharyngitis. The expression of known and putative virulence genes and regulatory genes (including genes in seven two-component regulatory systems) was studied. Several known and previously uncharacterized GAS virulence gene regulators were highly expressed compared to the constitutively expressed control gene proS. To examine in vivo gene transcription in a controlled setting, three cynomolgus macaques were infected with strain MGAS5005, an organism that is genetically representative of most serotype M1 strains recovered from pharyngitis and invasive disease episodes in North America and Western Europe. These three animals developed clinical signs and symptoms of GAS pharyngitis and seroconverted to several GAS extracellular proteins. Real-time RT-PCR analysis of throat swab material collected at intervals throughout a 12-day infection protocol indicated that expression profiles of a subset of GAS genes accurately reflected the profiles observed in the human pediatric patients. The results of our study demonstrate that analysis of in vivo GAS gene expression is feasible in throat swab specimens obtained from infected human and nonhuman primates. In addition, we conclude that the cynomolgus macaque is a useful nonhuman primate model for the study of molecular events contributing to acute pharyngitis caused by GAS. PMID- 12654844 TI - Cloning and characterization of an Ehrlichia canis gene encoding a protein localized to the morula membrane. AB - A gene encoding a 23.5-kDa ehrlichial morula membrane protein designated MmpA was cloned by screening an Ehrlichia canis expression library with convalescent dog sera, which resulted in three positive clones. Sequence analysis of the insert DNAs from all three clones indicated an open reading frame with a size of 666 bp that encodes MmpA. The structural analysis of MmpA indicated that it is a transmembrane protein with extreme hydrophobicity. Southern blot analysis of the HindIII-digested chromosomal DNA demonstrated the presence of a single copy of the mmpA gene in E. canis and Ehrlichia chaffeensis but not in the human granulocytic ehrlichiosis agent. The mmpA gene was amplified, cloned, and expressed as a fusion protein. Polyclonal antibodies to the recombinant protein (rMmpA) were raised in rabbits. Western blot analysis of E. canis and E. chaffeensis lysates with the anti-rMmpA serum resulted in the presence of an MmpA band only in E. canis, not in E. chaffeenesis. Sera from dogs which were either naturally or experimentally infected with E. canis recognized the recombinant protein. Double immunofluorescence confocal microscopy studies demonstrated that MmpA was localized mainly on the morula membrane of E. canis. Since the morula membrane is the interface between the ehrlichial growing environment and the host cytoplasm, MmpA may play a role in bacterium-host cell interactions. PMID- 12654845 TI - Passive immunity in Helicobacter-challenged neonatal mice conferred by immunized dams lasts until weaning. AB - The objective of this study was to examine the effect of breast-feeding by immunized dams on Helicobacter colonization in newborns. Urease-based immunization regimens failed to protect nursing pups against H. felis, whereas H. felis lysate-cholera toxin resulted in protection. This observation correlated with a high recognition of cell surface-expressed bacterial antigens by milk antibodies. Protection lasted until weaning, indicating that infection is maintained at undetectable levels by passive immunity but then resumes when breast-feeding stops. PMID- 12654843 TI - Caenorhabditis elegans as a model host for Staphylococcus aureus pathogenesis. AB - Staphylococcus aureus, an important pathogen of humans and other warm-blooded animals, is also capable of killing the nematode Caenorhabditis elegans. Here, we show that C. elegans organisms that are fed S. aureus die over the course of several days in a process that is correlated with the accumulation of bacteria within the nematode digestive tract. Several S. aureus virulence determinants known or speculated to be important in mammalian pathogenesis, including the quorum-sensing global virulence regulatory system agr and the global virulence regulator sarA, the alternative sigma factor sigma(B), alpha-hemolysin, and V8 serine protease, are required for full pathogenicity in nematodes. In addition, several defined C. elegans mutants were examined for susceptibility to S. aureus infection. Enhanced susceptibility to S. aureus killing was observed with loss-of function mutations in the C. elegans genes esp-2/sek-1 and esp-8/nsy-1, which encode components of a conserved p38 MAP kinase signaling pathway involved in nematode defense against multiple pathogens. These results suggest that key aspects of S. aureus pathogenesis have been conserved, irrespective of the host, and that specific C. elegans host factors can alter susceptibility to this gram positive human pathogen. PMID- 12654846 TI - Protein binding between PcrG-PcrV and PcrH-PopB/PopD encoded by the pcrGVH-popBD operon of the Pseudomonas aeruginosa type III secretion system. AB - Of the proteins encoded by the pcrGVH-popBD operon of the Pseudomonas aeruginosa type III secretion system, PcrG bound to PcrV and PcrH bound to PopB/PopD. In addition, Yersinia LcrG bound to PcrV, and Yersinia LcrH bound to PopD. The results imply a highly functional conservation of type III secretion between P. aeruginosa and Yersinia species. PMID- 12654847 TI - Synergistic protection of mice against plague with monoclonal antibodies specific for the F1 and V antigens of Yersinia pestis. AB - Monoclonal antibodies specific for Yersinia pestis V antigen and F1 antigen, administered singly or in combination, protected mice in models of bubonic and pneumonic plague. Antibodies showed synergy when administered prophylactically and as a therapy 48 h postinfection. Monoclonal antibodies therefore have potential as a treatment for plague. PMID- 12654848 TI - Enhanced murine antigen-specific gamma interferon and immunoglobulin G2a responses by using mycobacterial ESAT-6 sequences in DNA vaccines. AB - The Mycobacterium tuberculosis protein ESAT-6 has unusual immune stimulating activities, has been implicated in the recall of long-lived immunity, and induces protection against tuberculosis in mice. For many diseases caused by bacterial or viral pathogens, a strong cell-mediated immune (i.e., type 1) response is often required for recovery or protection. Therefore, it is important to design immunization regimens that induce agent-specific type 1 immunity. We have shown in previous studies that ESAT-6 could enhance antigen-specific type 1 immune responses in BALB/c mice against a second antigen when presented as a purified fusion protein. It was also of interest to determine if ESAT-6 could enhance the type 1 response against a second antigen beyond that afforded by DNA vaccination through CpG motifs. This was tested by using gene fusions of ESAT-6 and the Mycoplasma hyopneumoniae surface antigen P71. Modified P71 gene sequences were cloned with or without ESAT-6 sequences into a DNA vaccine vector and were used to immunize mice. Splenic lymphocytes from vaccinated mice were tested for gamma interferon (IFN-gamma) and interleukin-10 (IL-10) secretion. Serum antibodies were examined for P71 antigen-specific isotype responses. When stimulated in vitro with purified P71 antigen, splenocytes from the ESAT-6:P71 vaccinates secreted higher levels of IFN-gamma and lower levels of IL-10 compared to those of vaccinates receiving the P71 construct alone. Furthermore, the immunoglobulin G2a serum antibody levels were significantly higher in the ESAT-6:P71 vaccinates compared to those of the vaccinates receiving P71 alone. In conclusion, ESAT-6 was shown to enhance antigen-specific type 1 immune responses in BALB/c mice when used in DNA vaccines. PMID- 12654849 TI - Th2 responses predominate during the early phases of infection in patients with localized cutaneous leishmaniasis and precede the development of Th1 responses. AB - Intralesional Th2 responses preceded the development of Th1 responses in localized cutaneous leishmaniasis due to Leishmania guyanensis. Although the number of parasites increased in Th2 lesions, no correlation was found between the levels of cytokine expression and the number of parasites. In contrast, the decreased number of parasites in Th1 lesions is negatively correlated to gamma interferon expression. PMID- 12654850 TI - Molecular characterization of the prototrophic Salmonella mutants defective for intraepithelial replication. AB - Three MudJ prototrophs demonstrated that intracellular replication is a Salmonella virulence trait (K. Y. Leung and B. B. Finlay, Proc. Natl. Acad. Sci. USA, 88:11470-11474, 1991). mutS and mutH are disrupted in mutants 3-11 and 12 23, and ssaQ is disrupted in mutant 17-21. Further analysis revealed that loss of Salmonella pathogenicity island 2 function underlies the intracellular replication defect of 3-11 and 17-21. PMID- 12654851 TI - Neisseria meningitidis RTX proteins are not required for virulence in infant rats. AB - RTX cytotoxins play an important role in virulence of numerous gram-negative pathogens. Unexpectedly, however, we show here that the RTX proteins of Neisseria meningitidis are dispensable for virulence in the infant rat model of infection. PMID- 12654852 TI - Vector priming reduces the immunogenicity of Salmonella-based vaccines in Nramp1+/+ mice. AB - The present studies in Nramp1(-/-) BALB/c and Nramp1(+/+) CBA mice question the significance of this genotype as a determinant of the level of gut colonization following oral administration of naturally attenuated or highly virulent Salmonella strains. In line with previous results in BALB/c hosts, vector priming of CBA mice with Salmonella enterica serovar Stanley was found to significantly compromise the immunogenicity of a recombinant construct expressing a foreign pilus protein. PMID- 12654853 TI - Polymorphisms within EspA filaments of enteropathogenic and enterohemorrhagic Escherichia coli. AB - Enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC) possess a filamentous type III secretion system (TTSS) employed to deliver effector proteins into host cells. EspA is a type III secreted protein which forms the filamentous extension to the TTSS and which interacts with host cells during early stages of attaching and effacing (A/E) lesion formation. By immunofluorescence, a polyclonal antibody previously raised to EspA from EPEC strain E2348/69 (O127:H6) stained approximately 12-nm-diameter EspA filaments produced by this strain but did not stain similar filaments produced by EHEC serotype O157:H7. Similarly, an antibody that we subsequently raised to EHEC strain 85-170 (O157:H7) EspA stained approximately 12-nm-diameter EspA filaments produced by strain 85-170 but did not stain E2348/69 EspA filaments. Given such heterogeneity between EPEC and EHEC EspA filaments, we examined polymorphisms of functional EspA filaments among different EPEC and EHEC serotypes. With use of the EPEC EspA antiserum, EspA filaments were observed only with EPEC serotypes O127:H6 and O55:H6, serotypes which encode an identical EspA protein. When stained with the EHEC EspA antiserum, EspA filaments were detected only on EHEC strains belonging to serotype O157:H7; the EHEC antiserum did, however, stain EspA filaments produced by the closely related EPEC serotype O55:H7 but not filaments of any other EPEC serotype tested. Such polymorphisms among functional EspA filaments of EPEC and EHEC would be expected to have important implications for the development of broad-range EspA-based vaccines. PMID- 12654854 TI - Expression of Helicobacter pylori vacuolating toxin in Escherichia coli. AB - VacA is a secreted toxin that plays a role in Helicobacter pylori colonization of the stomach and that contributes to the pathogenesis of peptic ulcer disease. Studies of VacA structure and function have been hindered by the lack of an efficient system for expression and genetic manipulation of this toxin. In this study, we developed methodology for expression of a functionally active VacA toxin in Escherichia coli. We then used a high-throughput screen to analyze a library of mutant toxins with pentapeptide insertions and identified six mutants that lacked the capacity to induce vacuolation of HeLa cells. The capacity to analyze VacA in this heterologous-expression system should greatly facilitate efforts to elucidate the structure and function of this toxin. PMID- 12654855 TI - Mucosal immunization with a genetically engineered pertussis toxin S1 fragment cholera toxin subunit B chimeric protein. AB - A chimeric protein consisting of a divalent pertussis toxin (PT) S1 fragment linked to the cholera toxin (Ctx) A(2)B fragment was constructed. The chimera induced a mucosal immunoglobulin A (IgA) and a serum IgG immune response to PT and CtxB in BALB/c mice following intranasal immunization. The immune sera neutralized PT in vitro. In the mouse model of Bordetella pertussis respiratory infection, the chimera-immunized animals showed a significant reduction in bacterial lung counts (P = 0.01) from that of the sham control group. Thus, a divalent S1 fragment CtxA2B chimera is an immunogenic antigen and can elicit a protective immunity. PMID- 12654856 TI - Interleukin-10 negatively regulates local cytokine and chemokine production but does not influence antibacterial host defense during murine pneumococcal meningitis. AB - To determine the role of endogenous interleukin-10 (IL-10) in local host defense during pneumococcal meningitis, the inflammatory responses of IL-10-gene deficient and wild-type mice after the induction of meningitis were compared. The absence of IL-10 was associated with higher cytokine and chemokine concentrations and a more pronounced infiltrate, but antibacterial defense or survival was not influenced. PMID- 12654857 TI - Flagellum-mediated adhesion by Burkholderia pseudomallei precedes invasion of Acanthamoeba astronyxis. AB - In this study we investigated the role of the bacterial flagellum in Burkholderia pseudomallei entry to Acanthamoeba astronyxis trophozoites. B. pseudomallei cells were tethered to the external amoebic surface via their flagella. MM35, the flagellum-lacking fliC knockout derivative of B. pseudomallei NCTC 1026b did not demonstrate flagellum-mediated endocytosis in timed coculture, confirming that an intact flagellar apparatus assists B. pseudomallei entry into A. astronyxis. PMID- 12654858 TI - Immunization with Porphyromonas gingivalis capsular polysaccharide prevents P. gingivalis-elicited oral bone loss in a murine model. AB - The capsular polysaccharide (CPS) of the periodontal pathogen Porphyromonas gingivalis is an important virulence factor for this organism. We purified P. gingivalis CPS, immunized mice with this antigen, and assessed the vaccine potential of P. gingivalis CPS by using the murine oral challenge model. Animals immunized with P. gingivalis CPS developed elevated levels of immunoglobulin M (IgM) and IgG in serum that reacted with whole P. gingivalis organisms. The mice immunized with P. gingivalis CPS were protected from P. gingivalis-elicited oral bone loss. These data demonstrate that P. gingivalis CPS is a vaccine candidate for prevention of P. gingivalis-elicited oral bone loss. PMID- 12654859 TI - Bidirectional concentration-dependent effects of tumor necrosis factor alpha in Shigella dysenteriae-related seizures. AB - We have previously demonstrated that pretreatment of mice with Shigella dysenteriae sonicate enhanced their susceptibility to pentylenetetrazole-induced seizures and that tumor necrosis factor alpha (TNF-alpha) was proconvulsive in this respect. The present study shows that TNF-alpha, at high concentrations, may also exert a suppressive effect on Shigella-mediated seizures. This implies that high levels of TNF-alpha may play a protective role in neurologic complications of S. dysenteriae infection. PMID- 12654860 TI - The fibronectin-binding proteins of Staphylococcus aureus may promote mammary gland colonization in a lactating mouse model of mastitis. AB - The fibronectin-binding proteins (FnBPs) of Staphylococcus aureus are believed to be implicated in the pathogen's adherence to and colonization of bovine mammary glands, thus leading to infectious mastitis. In vitro studies have shown that FnBPs help the adhesion of the pathogen to bovine mammary epithelial cells. However, the importance of FnBPs for the infection of mammary glands has never been directly established in vivo. In this study with a mouse model of mastitis, the presence of FnBPs on the surface of S. aureus increased the capacity of the bacterium to colonize mammary glands under suckling pressure compared to that of a mutant lacking FnBPs. PMID- 12654862 TI - Why should we study the plant cell cycle? AB - Description of the molecular biology of plant and animal cell cycles highlights similarities and critical differences. The cell cycle is a point of control in both growth and development and deepening understanding of underlying processes and mechanisms may have many practical applications. PMID- 12654861 TI - Specificity and cross-reactivity of Plasmodium falciparum variant surface antigen specific antibody responses. PMID- 12654863 TI - Defining senescence and death. AB - This article evaluates features of leaf and flower senescence that are shared with, or are different from, those of other terminal events in plant development. Alterations of plastid structure and function in senescence are often reversible and it is argued that such changes represent a process of transdifferentiation or metaplasia rather than deterioration. It may be that the irreversible senescence of many flowers and some leaves represents the loss of ancestral plasticity during evolution. Reversibility serves to distinguish senescence fundamentally from programmed cell death (PCD), as does the fact that viability is essential for the initiation and progress of cell senescence. Senescence (particularly its timing and location) requires new gene transcription, but the syndrome is also subject to significant post- transcriptional and post-translational regulation. The reversibility of senescence must relate to the plastic, facultative nature of underlying molecular controls. Senescence appears to be cell-autonomous, though definitive evidence is required to substantiate this. The vacuole plays at least three key roles in the development of senescing cells: it defends the cell against biotic and abiotic damage, thus preserving viability, it accumulates metabolites with other functions, such as animal attractants, and it terminates senescence by becoming autolytic and facilitating true cell death. The mechanisms of PCD in plants bear a certain relation to those of apoptosis, and some processes, such as nucleic acid degradation, are superficially similar to aspects of the senescence syndrome. It is concluded that, in terms of physiological components and their controls, senescence and PCD are at best only distantly related. PMID- 12654864 TI - A novel alpha-helical protein, specific to and highly conserved in plants, is associated with the nuclear matrix fraction. AB - A cDNA for a novel plant protein was isolated from tomato. Nuclear Matrix Protein 1 (NMP1) is a ubiquitously expressed 36 kDa protein, which has no homologues in animals and fungi, but is highly conserved among flowering and non-flowering plants, including gymnosperms, moss, and the liverwort Marchantia polymorpha. NMP1 is predominantly alpha-helical with multiple stretches of short amphipathic regions. Cell fractionation, immunofluorescence, and GFP localization experiments showed that NMP1 is located both in the cytoplasm and nucleus and that the nuclear fraction is associated with the nuclear matrix. NMP1 is a candidate for a plant-specific structural protein with a function both in the nucleus and cytoplasm. PMID- 12654865 TI - Physiological, biochemical and molecular analysis of sugar-starvation responses in tomato roots. AB - Two-month-old tomato plants were submitted to day/night cycles and to prolonged darkness in order to investigate the physiological and biochemical response to sugar starvation in sink organs. Roots appeared particularly sensitive to the cessation of photosynthesis, as revealed by the reduction of the growth rate and the decline of the carbohydrate and protein content. Therefore, excised tomato roots were used as a model to deepen the characterization of sugar starvation symptoms. In excised roots, the endogenous sugars were rapidly exhausted and significant degradation of protein was observed. Glutamine and asparagine accounted for most of the nitrogen released by protein breakdown. Respiration declined and proliferation- and growth-associated genes were repressed soon after the beginning of the sugar depletion. Among the genes studied, only the gene encoding asparagine synthetase was strongly induced. All the starvation symptoms were reversible when the roots were resupplied with sugar. When the culture conditions deteriorated, the metabolic and molecular changes led to the triggering of apoptosis of the root cells. PMID- 12654866 TI - A jasmonate-responsive element within the A. thaliana vsp1 promoter. AB - The vsp1 gene of Arabidopsis thaliana encodes a storage protein that accumulates in vegetative organs. Transgenic plants expressing a vsp1 promoter-gus (beta glucuronidase) gene fusion were found to contain high GUS activity when challenged with jasmonate, a volatile plant hormone. The induction of vsp1-gus expression by jasmonate could be measured in tobacco mesophyll protoplasts, after transient expression. A number of deletions were operated in the vsp1 promoter in order to locate its jasmonate-responsive element. A 41 bp sequence taken approximately 150 bp upstream of the vsp1 TATA box could confer jasmonate responsiveness upon a short CaMV 35S promoter. Whereas the deletion of a CAAT box like element within the 41 bp sequence did not affect promoter activity, mutation of a short palindromic motif completely abolished jasmonate responsiveness. This motif shows no homology with the jasmonate-responsive elements of other promoters. PMID- 12654867 TI - Aluminium-responsive genes in sugarcane: identification and analysis of expression under oxidative stress. AB - Suppression subtractive hybridization (SSH) technology was used to gain preliminary insights into gene expression induced by the phytotoxic aluminium species, Al(3+), in sugarcane roots. Roots of hydroponically-grown Saccharum spp. hybrid cv. N19 were exposed to 221 microM Al(3+) at pH 4.1 for 24 h, a regime shown to inhibit root elongation by 43%, relative to unchallenged roots. Database comparisons revealed that, of a subset of 50 cDNAs ostensibly up-regulated by the metal in the root tips, 14 possessed putative identities indicative of involvement in signalling events and the regulation of gene expression, while the majority (28) were of unknown function. All of the 50 cDNAs sequenced displayed significant similarity to uncharacterized plant expressed sequence tags (ESTs), approximately half (23) of which had been derived from other graminaceous crop species that had been subject to a variety of stresses. Analysis of the expression of 288 putative Al(3+)-inducible genic fragments indicated higher levels of expression under oxidative (1 mM diamide for 4 h) rather than Al(3+) stress. By deploying SSH, this study has provided an indication of the nature of genes expressed in sugarcane roots under Al(3+) stress. It is anticipated that the information obtained will guide further exploration of the potential for manipulation of the Al tolerance characteristics of the crop. PMID- 12654868 TI - Isolation of tobacco ubiquitin-conjugating enzyme cDNA in a yeast two-hybrid system with tobacco ERF3 as bait and its characterization of specific interaction. AB - Tobacco ETHYLENE-RESPONSIVE FACTOR3 (ERF3) is a member of the ERF-domain transcription factors and has a transcriptional repressor activity, whereas other ERF proteins show activation activity. To understand the regulation of ERF3 repressor activity, protein(s) were screened which interact with ERF3 in a yeast two-hybrid system. A partial sequence (B8) of NtUBC2, a tobacco ubiquitin conjugating enzyme was isolated. This B8 specifically interacted with ERF3 in the yeast two-hybrid system. Further analyses revealed that the region unique to ERF3 interacted with B8. The physiological functions of NtUBC2 and the stability of ERF3 are discussed in relation to the regulation of the repression activity of ERF3. PMID- 12654869 TI - Isolation and characterization of a cDNA encoding a lipid transfer protein expressed in 'Valencia' orange during abscission. AB - The genetics and expression of a lipid transfer protein (LTP) gene was examined during abscission of mature fruit of 'Valencia' orange. A cDNA encoding an LTP, CsLTP, was isolated from a cDNA subtraction library constructed from mature fruit abscission zones 48 h after application of a mature fruit-specific abscission agent, 5-chloro-3-methyl-4-nitro-pyrazole (CMN-pyrazole). A full-length cDNA clone of 652 nucleotides was isolated using 5' and 3' RACE followed by cDNA library screening and PCR amplification. The cDNA clone encoded a protein of 155 amino acid residues with a molecular mass and isoelectric point of 9.18 kDa and 9.12, respectively. A partial genomic clone of 505 nucleotides containing one intron of 101 base pairs was amplified from leaf genomic DNA. Southern blot hybridization demonstrated that at least two closely related CsLTP genes are present in 'Valencia' orange. Temporal expression patterns in mature fruit abscission zones were examined by northern hybridization. Increased expression of CsLTP mRNA was detected in RNA of mature fruit abscission zones 6, 24, 48, and 72 h after application of a non-specific abscission agent, ethephon. Low expression of CsLTP transcripts was observed after treatment of CMN-pyrazole until 24 h after application. After this time, expression markedly increased. The results suggest that CsLTP has a role in the abscission process, possibly by assisting transport of cutin monomers to the fracture plane of the abscission zone or through its anti-microbial activity by reducing the potential of microbial attack. PMID- 12654870 TI - Expression of a putative grapevine hexose transporter in tobacco alters morphogenesis and assimilate partitioning. AB - Tobacco plants were transformed by leaf disc regeneration with the VvHT1 (Vitis vinifera hexose transporter 1) cDNA under the control of the constitutive CaMV 35S promoter in a sense or antisense orientation. Among the 20 sense plants and 10 antisense plants obtained, two sense plants showed a mutant phenotype when grown in vitro, with stunted growth and an increase in the (leaves+stem)/roots dry weight ratio. The rate of [(3)H]-glucose uptake in leaf discs from these plants was decreased to 25% of the value measured in control plants. The amount of VvHT1 transgene and of host monosaccharide transporter MST transcripts in the leaves were studied by RNA gel blot analysis. The VvHT1 transcripts were usually present, but the amount of MST transcripts was the lowest in the plants that exhibited the most marked phenotype. Although the phenotype was lost when the plants were transferred from in vitro to greenhouse conditions, it was found again in vitro in the progeny obtained by self-pollination or by back-cross. The data show that VvHT1 sense expression resulted in unidirectional post transcriptional gene inactivation of MST in some of the transformants, with dramatic effects on growth. They provide the first example of plants modified for hexose transport by post-transcriptional gene silencing. Some of the antisense plants also showed reduced expression of MST, and decreased growth. These results indicate that, like the sucrose transporters, hexose transporters play an important role in assimilate transport and in morphogenesis. PMID- 12654871 TI - Non-hydraulic regulation of fruit growth in tomato plants (Lycopersicon esculentum cv. Solairo) growing in drying soil. AB - Tomato (Lycopersicon esculentum cv. Solairo) fruit growth, fruit mesocarp and leaf epidermal cell turgor, and fruit and leaf sub-epidermal apoplastic pH were monitored as plants were allowed to dry the soil in which they were rooted. Soil drying regimes involved splitting the root system of plants between two halves of a single pot separated by a solid impervious membrane to form a split-root system. Plants were then allowed to dry the soil in both halves of the pot (a soil-drying (SD) treatment) or water was supplied to one-half of the pot (a partial root-drying (PRD) treatment), allowing only one-half of the root system to dry the soil. A well-watered control treatment watered the soil on both halves of the pot. The rate of fruit growth was highly correlated with the soil water content of both sides of the SD treatment and the dry side of the PRD treatment. Soil drying caused a significant restriction in fruit growth rate, which was independent of any changes in the turgor of expanding fruit mesocarp cells in the PRD treatment. By supplying water to half of the root system, the turgors of mesocarp cells were maintained at values above those recorded in well-watered controls. The turgor of leaf epidermal cells exhibited a similar response. The pH of the sub-epidermal apoplastic compartment in leaves and fruit increased with soil drying. The dynamics of this increase in leaves and fruit were identical, suggesting free transport of this signal from shoot to fruit. Fruit growth rate and sub-epidermal pH within the fruit showed a strong correlation. The similarity of fruit growth response in the SD and PRD treatment, suggests that tomato plants respond to a discrete measure of soil water status and do not integrate measures to determine total soil water availability. The results of this study are not consistent with Lockhartian models of growth regulation in expanding fruit of a higher plant. A non-hydraulic, chemical-based signalling control of fruit growth in plants growing in drying soil is proposed. PMID- 12654872 TI - Effects of the experimental blockage of the major veins on hydraulics and gas exchange of Prunus laurocerasus L. leaves. AB - The impact of leaf vein blockage on leaf hydraulic conductance (K(L)), gas exchange (g(L)) and water potential (Psi(L)) was studied in Prunus laurocerasus L., a broad-leaved evergreen. For this purpose, leaves were measured for the three variables above, either with an intact leaf blade (controls) or with the midrib cut a third of the way up (cut a), or with the midrib cut at three different points and the first-order veins cut through near their insertion to the midrib (cut b), or with the midrib cut at 2 mm from the leaf base (cut c). All the cut surfaces were sealed with cyanoacrylate. A serial decrease of K(L) was recorded from cut a to cut c with respect to that measured for the controls, i.e. a K(L) loss of about 37% (cut a), 57% (cut b) and 87% (cut c). A positive linear relationship appeared to exist between g(L) and K(L) with a high correlation coefficient (r(2)=0.99) and a high statistical significance (P <0.01). Even under a severe drop in K(L) (as that induced by cut c), leaf water potential remained approximately constant and not statistically different from Psi(L) measured for the controls. In fact, Psi(L) ranged between -0.83 and -0.98 MPa, i.e. within the cavitation threshold of leaves in terms of the critical Psi(L) inducing a significant production of ultrasound acoustic emissions which was -0.94+/-0.09 MPa. The conclusion was that stomata were very sensitive to changes in K(L) and that stomatal closure led to the homeostatic maintenance of Psi(L) and cavitation avoidance. PMID- 12654873 TI - Root and stem hydraulic conductivity as determinants of growth potential in grafted trees of apple (Malus pumila Mill.). AB - The anatomy of the graft tissue between a rootstock and its shoot (scion) can provide a mechanistic explanation of the way dwarfing Malus rootstocks reduce shoot growth. Considerable xylem tissue disorganization may result in graft tissue having a low hydraulic conductivity (k(h)), relative to the scion stem. The graft may influence the movement of substances in the xylem such as ions, water and plant-growth-regulating hormones. Measurements were made on 3-year-old apple trees with a low-pressure flow system to determine k(h) of root and scion stem sections incorporating the graft tissue. A range of rootstocks was examined, with different abilities of dwarfing; both ungrafted and grafted with the same scion shoot cultivar. The results showed that the hydraulic conductivity (k(hroot)) of roots from dwarfing rootstocks was lower compared with semi vigorous rootstocks, at least for the size class of root measured (1.5 mm diameter). Scion hydraulic conductivity (k(hs)) was linked to leaf area and also to the rootstock on to which it was grafted, i.e. hydraulic conductivity was greater for the scion stem on the semi-vigorous rootstock. Expressing conductivities relative to xylem cross-sectional areas (k(s)) did not remove these differences suggesting that there were anatomical changes induced by the rootstock. The calculated hydraulic conductivity of the graft tissue was found to be lower for grafted trees on dwarfing rootstocks compared to invigorating rootstocks. These observations are discussed in relation to the mechanism(s) by which rootstock influences shoot growth in grafted trees. PMID- 12654874 TI - Fate of fructose supplied to leaf sheaths after defoliation of Lolium perenne L.: assessment by 13C-fructose labelling. AB - The role of fructans from leaf sheaths for the refoliation of Lolium perenne after severe defoliation was assessed by following the fate of (13)C-fructose supplied to leaf sheaths at the time of defoliation. At the end of the 4 h labelling period on defoliated plants, 77% of the (13)C incorporated was still located in leaf sheaths. Only 4% and 0.9% were, respectively, allocated to stem and roots, while 18% was imported by the growing leaves where (13)C was allocated first to the proximal part of the leaf growth zone (0-10 mm). In all tissues, the most highly (13)C-labelled carbohydrates was not fructose but sucrose. In leaf sheaths, (13)C-loliose was produced. In the leaf growth zone (0-20 mm), fructans were simultanously synthesized from (13)C entering the leaves and degraded. The export of (13)C from leaf sheaths continued during the first day of regrowth but stopped afterwards. There was no net loss of C from (13)C-fructose over the first 2 d of regrowth. The role of fructans and loliose is discussed as well as the physiological mechanisms contributing to defoliation tolerance in L. perenne. PMID- 12654875 TI - Expression of CycD3 is transiently increased by pollination and N-(2-chloro-4 pyridyl)-N'-phenylurea in ovaries of Lagenaria leucantha. AB - Lagenaria leucantha is an important vegetable crop and a potential model for the study of fruit development. To study the function of D cyclins in fruit development, full-length cDNA clones for two D cyclin genes were isolated from young ovaries of Lagenaria leucantha. They were classified as D3 cyclins by sequence similarities and phylogenetic analysis, and nominated LlCycD3;1 and LlCycD3;2, respectively. The deduced amino acid sequence of both LlCycD3 genes contained a retinoblastoma-binding motif and a PEST-destruction motif. Unpollinated ovaries failed to develop and eventually aborted. N-(2-chloro-4 pyridyl)-N'-phenylurea (CPPU) induced parthenocarpic fruit significantly larger than pollinated ones. In unpollinated ovaries, the expression of both LlCycD3 genes was abundant at anthesis and then suddenly decreased, concomitant with the cessation of cell division. Pollination/fertilization induced an activation of the cell cycle accompanied by a large increase in the transcript levels of LlCycD3;1 and LlCycD3;2 in young fruits. Treating ovaries with CPPU also reactivated cell division and transcription of CycD3 genes and the effect was more rapid and pronounced than after pollination/fertilization. PMID- 12654876 TI - Regulation of nodulation in the absence of N2 is different in actinorhizal plants with different infection pathways. AB - Root nodulation in actinorhizal plants, like Discaria trinervis and Alnus incana, is subject to feedback regulatory mechanisms that control infection by Frankia and nodule development. Nodule pattern in the root system is controlled by an autoregulatory process that is induced soon after inoculation with Frankia. The final number of nodules, as well as nodule biomass in relation to plant biomass, are both modulated by a second mechanism which seems to be related to the N status of the plant. Mature nodules are, in part, involved in the latter process, since nodule excision from the root system releases the inhibition of infection and nodule development. To study the effect of N(2) fixation in this process, nodulated D. trinervis and A. incana plants were incubated under a N(2)-free atmosphere. Discaria trinervis is an intercellularly infected species while A. incana is infected intracellularly, via root hairs. Both symbioses responded with an increment in nodule biomass, but with different strategies. Discaria trinervis increased the biomass of existing nodules without significant development of new nodules, while in A. incana nodule biomass increased due to the development of nodules from new infections, but also from the release of arrested infections. It appears that in D. trinervis nodules there is an additional source for inhibition of new infections and nodule development that is independent of N(2) fixation and nitrogen assimilation. It is proposed here that the intercellular Frankia filaments commonly present in the D. trinervis nodule apex, is the origin for the autoregulatory signals that sustain the blockage of initiated nodule primordia and prevent new roots from infections. When turning to A. incana plants, it seems likely that this signal is related to the early autoregulation of nodulation in A. incana seedlings and is no longer present in mature nodules. Thus, actinorhizal symbioses belonging to relatively distant phylogenetic groups and displaying different infection pathways, show different feedback regulatory processes that control root nodulation by Frankia. PMID- 12654877 TI - Characterization and genetic analysis of a lettuce (Lactuca sativa L.) mutant, weary, that exhibits reduced gravitropic response in hypocotyls and inflorescence stems. AB - A lettuce (Lactuca sativa L.) mutant that exhibits a procumbent growth habit was identified and characterized. In two wild type (WT) genetic backgrounds, segregation patterns revealed that the mutant phenotype was controlled by a recessive allele at a single locus, which was designated weary. Hypocotyls and inflorescence stems of plants homozygous for the weary allele exhibited reduced gravitropic responses compared with WT plants, but roots exhibited normal gravitropism. Microscopic analysis revealed differences in the radial distribution of amyloplasts in hypocotyl and inflorescence stem cells of weary and WT plants. Amyloplasts occurred in a single layer of endodermal cells in WT hypocotyls and inflorescence stems. By contrast, amyloplasts were observed in several layers of cortical cells in weary hypocotyls, and weary inflorescence stem cells lacked amyloplasts entirely. These results are consistent with the proposed role of sedimenting amyloplasts in shoot gravitropism of higher plants. The phenotype associated with the weary mutant is similar to that described for the Arabidopsis mutant sgr1/scr, which is defective in radial patterning and gravitropism. PMID- 12654878 TI - Sodium chloride reduces growth and cytosolic calcium, but does not affect cytosolic pH, in root hairs of Arabidopsis thaliana L. AB - The effects of salinity (NaCl) stress on growth, cytosolic Ca(2+) gradients and cytosolic pH homeostasis of root hairs of Arabidopsis thaliana are assessed here. Neither cytosolic Ca(2+) nor pH at the hair apex were significantly affected by 20 min exposure of up to 90 mM NaCl or of up to 5 mM extracellular Ca(2+). Exposure to increasing NaCl concentrations, up to 90 mM, for 2 d or 6 d reduced hair extension, and this inhibition was relieved by supplemental extracellular Ca(2+). Such extended salinity stress reduced the magnitude of the Ca(2+) gradient in the apical 12 microm of hairs at all NaCl concentrations tested (up to 90 mM), including NaCl concentrations that did not reduce hair extension. The magnitude of the tip-focused gradient was also reduced in root hairs of plants grown with low (0.5 mM) extracellular Ca(2+) when compared to those in 5 mM extracellular Ca(2+), regardless of the presence of NaCl. Up to 90 mM NaCl did not affect cytosolic pH of root hairs in any of the treatments. It is concluded that NaCl inhibition of root hair extension in the long term may operate via alterations in the tip-focused Ca(2+) gradient that regulates root hair growth. However, NaCl-induced alterations in this gradient do not always lead to detectably altered growth kinetics. Short-term signalling events in response to NaCl may operate by a means other than altering Ca(2+) at the root hair apex. Salinity stress in root hairs does not appear to be mediated by effects on cytosolic pH. PMID- 12654879 TI - Do increases in xylem sap pH and/or ABA concentration mediate stomatal closure following nitrate deprivation? AB - Stomatal conductance (g(s)) of pepper (Capsicum annuum L.) plants decreased during the second photoperiod (day 2) after withholding nitrate (N). Stomatal closure of N-deprived plants was not associated with a decreased shoot water potential (Psi(shoot)); conversely Psi(shoot) was lower in N-supplied plants. N deprivation transiently (days 2 and 3) alkalized (0.2-0.3 pH units) xylem sap exuded from de-topped root systems under root pressure, and xylem sap expressed from excised shoots by pressurization. The ABA concentration of expressed sap increased 3-4-fold when measured on days 2 and 4. On day 2, leaves detached from N-deprived and N-supplied plants showed decreased transpiration rates when fed an alkaline (pH 7) artificial xylem (AX) solution, independent of the ABA concentration (10-100 nM) supplied. Thus changes in xylem sap composition following N deprivation can potentially close stomata. However, the lower transpiration rate of detached N-deprived leaves relative to N-supplied leaves shows that factors residing within N-deprived leaves also mediate stomatal closure, and that these factors assume greater importance as the duration of N deprivation increases. PMID- 12654881 TI - Quantitative assessment to the structural basis of water repellency in natural and technical surfaces. AB - Many plant surfaces are water-repellent because of a complex 3-dimensional microstructure of the epidermal cells (papillae) and a superimposed layer of hydrophobic wax crystals. Due to its surface tension, water does not spread on such surfaces but forms spherical droplets that lie only on the tips of the microstructures. Studying six species with heavily microstructured surfaces by a new type of confocal light microscopy, the number, height, and average distance of papillae per unit area were measured. These measurements were combined with those of an atomic force microscope which was used to measure the exposed area of the fine-structure on individual papillae. According to calculations based upon these measurements, roughening results in a reduction of the contact area of more than 95% compared with the projected area of a water droplet. By applying water/methanol solutions of decreasing surface tension to a selection of 33 water repellent species showing different types of surface structures, the critical value at which wetting occurs was determined. The results impressively demonstrated the importance of roughening on different length scales for water repellency, since extremely papillose surfaces, having an additional wax layer, are able to resist up to 70% methanol. Surfaces that lack papillae or similar structures on the same length scale are much more easily wetted. PMID- 12654880 TI - Fatty acid elongation is important in the activity of thiocarbamate herbicides and in safening by dichlormid. AB - The thiocarbamates, such as pebulate (S-propyl butyl (ethyl) thiocarbamate) are a well-established class of herbicides. They inhibit fatty acid elongation, which is necessary for the biosynthesis of constituents of surface waxes and suberin and this has been proposed to be important for their toxicity. In this study lipid metabolism was investigated in herbicide-treated barley (Hordeum vulgare) and a pernicious weed, wild oats (Avena ludoviciana), to test the hypothesis that inhibitory effects on fatty acid elongation could be counteracted by the safer, dichlormid. Pebulate and its sulphoxide derivative (thought to be the active metabolite in vivo) were tested against lipid metabolism in barley or wild oat shoots. In both plants there was a significant inhibition of very long chain fatty acid (VLCFA) synthesis at herbicide concentrations > or =25 micro M. The extent to which safener dichlormid could prevent the inhibition of VLCFA synthesis was different in the two species. Previous treatment of barley with dichlormid (N,N-diallyl-2,2-dichloroacetamide) enabled fatty acid elongation in the presence of pebulate or pebulate sulphoxide, but had no effect on wild oats. The effects on fatty acid elongation mimicked the differential safening action of dichlormid observed on shoot elongation and growth in the two species. These data provide further evidence that inhibition of VLCFA formation is important for the mechanism of action of thiocarbamates. PMID- 12654882 TI - Resistance to broomrape (Orobanche spp.) in sunflower (Helianthus annuus L.) is temperature dependent. AB - The effects of various temperature regimes in the range 29-17/21-9 degrees C day/night on each stage of the parasitism process of Orobanche cumana and O. aegyptiaca on sunflower were studied under controlled conditions in polyethylene bags. The response of the resistant sunflower variety 'Ambar' was expressed as the degeneration of the parasite tissues after its establishment in the plant roots, and this stage was found to be temperature dependent. The degeneration rate of Orobanche tubercles in the resistant sunflower variety was also found to be temperature dependent and was about five times as great as that in the sensitive variety in the highest temperature regime tested of 29/21 degrees C day/night. The ability to reject the parasite by causing its degeneration and death is the main factor that determines the resistance. As the temperature rises, more tubercles degenerate and die, that is the sunflower plant expresses higher levels of resistance. PMID- 12654883 TI - Insights into the evolution of human bipedalism from experimental studies of humans and other primates. AB - An understanding of the evolution of human bipedalism can provide valuable insights into the biomechanical and physiological characteristics of locomotion in modern humans. The walking gaits of humans, other bipeds and most quadrupedal mammals can best be described by using an inverted-pendulum model, in which there is minimal change in flexion of the limb joints during stance phase. As a result, it seems logical that the evolution of bipedalism in humans involved a simple transition from a relatively stiff-legged quadrupedalism in a terrestrial ancestor to relatively stiff-legged bipedalism in early humans. However, experimental studies of locomotion in humans and nonhuman primates have shown that the evolution of bipedalism involved a much more complex series of transitions, originating with a relatively compliant form of quadrupedalism. These studies show that relatively compliant walking gaits allow primates to achieve fast walking speeds using long strides, low stride frequencies, relatively low peak vertical forces, and relatively high impact shock attenuation ratios. A relatively compliant, ape-like bipedal walking style is consistent with the anatomy of early hominids and may have been an effective gait for a small biped with relatively small and less stabilized joints, which had not yet completely forsaken arboreal locomotion. Laboratory-based studies of primates also suggest that human bipedalism arose not from a terrestrial ancestor but rather from a climbing, arboreal forerunner. Experimental data, in conjunction with anatomical data on early human ancestors, show clearly that a relatively stiff modern human gait and associated physiological and anatomical adaptations are not primitive retentions from a primate ancestor, but are instead recently acquired characters of our genus. PMID- 12654884 TI - Localization of myoinhibitory peptide immunoreactivity in Manduca sexta and Bombyx mori, with indications that the peptide has a role in molting and ecdysis. AB - For normal development of Manduca sexta larvae, the ecdysteroid titer must drop following its sudden rise at the start of the molting cycle; this sudden decline in titer may be due to myoinhibitory peptide I (MIP I), which has an inhibitory effect on the release of ecdysone by the prothoracic glands of Bombyx mori in vitro. Using an antiserum to MIP, we have demonstrated secretion of an MIP-like peptide by the epiproctodeal glands of Manduca sexta, which are located on each proctodeal nerve, just anterior to the rectum. These MIP-immunoreactive glands are also present in B. mori. In fourth-instar larvae of M. sexta, the epiproctodeal glands show a distinct cycle of synthesis and sudden release of MIP that coincides with the time of the rapid decline in ecdysteroid titer. The function of the epiproctodeal glands appears to be the timely release of MIP during the molting cycle, so as to inhibit the prothoracic glands and thus to facilitate the sudden decline in ecdysteroid titer. In addition, we have found that MIP immunoreactivity is co-localized with that of crustacean cardioactive peptide (CCAP) in the 704 interneurons; these peptides appear to be co-released at the time of ecdysis. It is known that CCAP can initiate the ecdysis motor program; our results suggest that MIP may also be involved in activating ecdysis behavior. PMID- 12654885 TI - Factors affecting stroking patterns and body angle in diving Weddell seals under natural conditions. AB - Aquatic animals use a variety of strategies to reduce the energetic cost of locomotion. Efficient locomotion is particularly important for breath-holding divers because high levels of exercise may quickly deplete oxygen reserves, leading to the termination of a dive. We investigated the swimming behavior of eight adult Weddell seals, which are proficient divers, in McMurdo Sound, Antarctica. A newly developed data logger was attached to free-ranging females at their own breeding sites to record swimming speed, depth, two-dimensional accelerations (stroke frequency and body angle) and temperature. All seals conducted multiple deep dives (the mean dive depth range for each animal was 223.3+/-66.5-297.9+/-164.7 m). Prolonged gliding while descending was observed with thinner females (N=5 seals). But the fatter females (N=3 seals) exhibited only swim-and-glide swimming, characterized by intermittent stroking and fluctuating swim speed, throughout their descent and ascent. The body angles of four of the seals were restricted to less than 30 degrees by the location of breathing holes in the ice and the slope of local bathymetric features. Of these four, the three fatter seals adopted the stroke-and-glide method while the other thinner seal descended with prolonged periods of gliding. Prolonged gliding seems to be a more efficient method for locomotion because the surface time between dives of gliding seals was significantly less than that of stroking animals, despite their same stroke frequencies. PMID- 12654886 TI - Heterothermy and the water economy of free-living Arabian oryx (Oryx leucoryx). AB - To test the idea that large, free-living, desert ungulates use heterothermy to reduce water loss, we measured core body temperature (T(b)) of six free-ranging, adult Arabian oryx (Oryx leucoryx) during 2 years in the arid desert of west central Saudi Arabia. We report the first case of heterothermy in a free-living ruminant in a desert environment: T(b) varied by 4.1+/-1.7 degrees C day(-1) during summer (June to September) and by 1.5+/-0.6 degrees C day (-1) during winter (November to March). Over both seasons, mean T(b) was 38.4+/-1.3 degrees C. During the day in both summer and winter, T(b) increased continually, suggesting that oryx store heat instead of dissipating it by evaporation, whereas at night T(b) decreased. The minimum T(b) was lower in summer (36.5+/-1.16 degrees C) than in winter (37.5+/-0.51 degrees C) despite the fact that the temperature gradient between T(b) and air temperature (T(a)) was larger and solar radiation was lower in winter. Throughout the year, daily variation in T(b) appeared to reflect thermal load (T(a,max)-T(a,min)) rather than an endogenous rhythm. Behavioural thermoregulation was used by oryx to cope with thermal stress during summer: animals lay down in shade in the morning shortly before T(a) exceeded T(b) and remained there until evening when T(b)-T(a) became positive. The use of heterothermy by oryx resulted in storage of 672.4 kJ day(-1) animal( 1) in summer and 258.6 kJ day(-1) animal(-1) in winter, if heat storage is based on calculations involving mean T(b). To dissipate this heat by evaporation would require 0.28 litres H(2)O day(-1) animal(-1) and 0.11 litres H(2)O day(-1) animal(-1) in summer and winter, respectively. Without heat storage in summer, we estimated that oryx would have to increase their water intake by 19%, a requirement that would be difficult to meet in their desert environment. If heat storage was calculated based on the daily change in T(b) rather than on heat storage above mean T(b) then we estimated that oryx saved 0.538 litres H(2)O day( 1) animal(-1) during summer. PMID- 12654887 TI - Wing resonances in the Australian field cricket Teleogryllus oceanicus. AB - The anatomy and mechanics of the fore-wings of the Australian cricket Teleogryllus oceanicus were examined to study how resonances of the wings were excited, to model the interactions between the two wings during sound production, to account for the frequency changes that occur within the pulses and to determine the variation in sound amplitude during the pulses. Sound is produced after raising the wings by closing the right wing over the left; the plectrum of the left wing engages and releases teeth on the file on the underside of the right wing. The mean number of teeth on the right file is 252; the teeth are more closely spaced in the posterior part of the file, which is engaged at the start of the song pulses. The anterior part of the file is separated from the base of the harp by a short flexible region. The dorsal field of the wing, in which the harp is situated, is largely mechanically isolated from the driving veins of the lateral field, except for a cross vein at the apex of the harp. The harps of the two wings did not differ significantly in area but the plectrum of the left wing was significantly longer and wider than that of the right wing. The posterior edge of the plectrum has a radius of approximately 0.5 micro m, which allows it to engage the 20 micro m-tall teeth of the file. The plectrum is separated from the wing by a 0.5 micro m-thick crescent that allows it to twist lengthways and thus disengage the file teeth. The sigmoid shape of the file allows the plectrum to engage teeth over most of the length of the file. The calling song of T. oceanicus consists of a chirp of four similar pulses followed by a trill of pairs of pulses. The dominant frequency of all pulses is approximately 4.8 kHz but cycle-by-cycle analysis suggests that the different types of pulse are produced by wing-closing movements through different arcs. Free resonances of the left wing occurred at 4.56 kHz [quality factor (Q)=25.1] and of the right wing at 4.21 kHz (Q=23.9). Driven by loud sound, maximum vibration of the harp was seen at approximately 4.5 kHz; at lower sound levels, the vibration was confined to the cross-veins of the harp that extend distally from the file. Resonances of the left wing driven by vibration of the same wing, either at the plectrum or on the anal area, occurred at similar frequencies to those of the songs and had similar Qs but were approximately anti-phase, demonstrating that movement of the plectrum (e.g. by the file teeth) causes an opposite movement of the harp. When the right wing was driven directly on the file, the resonant frequency was 5.88 kHz but, when driven on the file via a length of the left file and the left plectrum, it was 4.83 kHz. The amplitude of the vibration increased from the posterior end of the file to the middle then fell towards the anterior end of the file. Pushing a left plectrum across the middle of the right file produced trains of damped sound pulses at 4.82 kHz (Q=23.4). Clicks excited from the anterior end of the file had lower frequencies. The resonances excited from both the left wing via its plectrum and from the right wing when driven via the left plectrum were similar in frequency to that of the song. The resonance of the dorsal field persisted after ablation of the harp but the mean resonant frequency increased 1.12-fold with a similar Q to the intact wing. Droplets of water on the distal end of the harp or proximal part of the dorsal field raised the resonant frequency. The resonant frequency was lowered by the addition of weights to the harp or the file; the factor of the decrease suggested that the mass of the resonant system was approximately 1.4 mg, which accords with the mass of the harp plus file plus anal area of the wing (left wing, 1.27 mg; right wing, 1.15 mg) but is far heavier than the harp (0.22 mg). An earlier suggestion that the harp is the resonator is not supported; instead, it is proposed that the major elastic component of the resonant system is the file plus 1st anal vein and that the mass component is the combined mass of the file, anal area and harp. PMID- 12654888 TI - Behavioral attraction of Leach's storm-petrels (Oceanodroma leucorhoa) to dimethyl sulfide. AB - A recent model for olfactory foraging by procellariiform seabirds suggests that these birds use biogenic sulfur compounds to locate productive areas for foraging in the southern oceans. The present study refines a simple approach to test birds' responses to odors on land and extends our knowledge to a northern species, the Leach's storm-petrel (Oceanodroma leucorhoa). Rather than working at sea, we tested the behavioral response to dimethyl sulfide (DMS) at night in breeding colonies on Kent Island, New Brunswick, Canada. Birds were presented with either 5 ml DMS (100 micro mol l(-1) concentration) or control (water) solutions from a platform 1.5 m in height positioned in a flyway 10 m from a breeding colony. We also tested birds' responses to cod liver oil, a well established olfactory attractant of procellariiforms foraging at sea. Leach's storm-petrels approached DMS presentations nearly twice as frequently as they approached controls. We next compared the distribution of approaches against a Poisson process to test for evidence of social cueing. We found that approaches to DMS were significantly clustered. By contrast, the distribution of approaches did not depart significantly from a Poisson distribution for either cod liver oil or control presentations. Taken together, these results suggest that Leach's storm-petrels can smell DMS and potentially use it as a foraging cue. The results are consistent with the hypothesis that the detection of biogenic sulfur compounds in combination with other cues assists birds in locating foraging hotspots. PMID- 12654889 TI - Regulation of Na+/K+-ATPase activity by nitric oxide in the kidney and gill of the brown trout (Salmo trutta). AB - In teleost fish, successful osmoregulation involves controlled ion transport mechanisms in kidney and gill epithelia. In this study, the effect of nitric oxide (NO) on Na(+)/K(+)-ATPase was investigated in vitro in these two tissues in brown trout (Salmo trutta) acclimated to freshwater. The NO donor sodium nitroprusside (SNP) inhibited in situ Na(+)/K(+)-ATPase activity, measured as ouabain-sensitive Rb(+) uptake, in both samples of kidney and gill tissue and in isolated gill cells. The effect was dose-dependent in both tissues, with a maximal observed inhibition of approximately 40-50% (1 mmol l (-1) SNP). The time course of inhibition revealed a maximum effect with 10 min pre-incubation. The effect of SNP was reproduced with another NO donor, papa-nonoate (NOC-15; 200 micro mol l(-1)), and was prevented by the NO scavenger 2-phenyl-4,4,5,5 tetramethylimidazoline-1-oxyl 3-oxide (PTIO; 1 mmol l(-1)). To further investigate the mechanism of the NO effect, whole-tissue Na(+) and K(+) levels were analysed. In kidney, SNP (1 mmol l(-1)) led to an increase in tissue Na(+) levels and a decrease in K(+) levels in a 3:2 ratio. In gill tissue, no change in either ion was observed. These observations indicate that the effect on Na(+)/K(+)-ATPase is direct rather than due to a decrease in intracellular Na(+), its rate-limiting substrate. SNP elevated the level of cyclic GMP (cGMP) in both kidney and gill tissue. Dibutyryl cyclic GMP (db-cGMP; 1 mmol l(-1)) also inhibited Na(+)/K(+)-ATPase activity in both tissues. Hence, a possible mechanism may involve the cGMP-activated kinase, even though other mechanisms cannot be excluded. PMID- 12654890 TI - Evolution of glutamine synthetase in vertebrates: multiple glutamine synthetase genes expressed in rainbow trout (Oncorhynchus mykiss). AB - Glutamine synthetase (GSase) is a key enzyme in nitrogen metabolism and encoded by a single gene in mammals. Using PCR cloning techniques, including RT-PCR from total RNA and PCR from a cDNA library, we find evidence of four expressed GSase mRNAs for the tetraploid rainbow trout. For two of these mRNAs (Onmy-GS01, -GS02) we characterize the full-length coding regions, and for two others (Onmy-GS03, GS04), we describe partial sequences. Northern analysis of Onmy-GS01, -GS02, GS03 and -GS04 indicates that (1) Onmy-GS02 is expressed at higher levels relative to the other transcripts in most adult tissues, with the exception of brain and gill, where Onmy-GS01 is at the highest level, and (2) the tissue with the highest level of expression of all four transcripts is the brain, with decreasing levels in the intestine, liver, red muscle, gill/kidney, white muscle and heart. Clearly, rainbow trout possess multiple GSase genes with differing levels of tissue expression, implying manifold potential routes of regulation for this octameric enzyme. Our data also indicate that caution should be taken when interpreting mRNA expression data of a single gene, unless multiple genes have been ruled out. Consistent with a southern blot, phylogenetic and intron sequence analyses imply that the trout genes are encoded by at least four separate loci, belonging to two distinct evolutionary branches. Our data on rainbow trout, together with those from two full-length zebrafish Danio rerio GSase genes compiled from GenBank ESTs, support the idea that fish GSases are polyphyletic and that gene duplications have occurred at multiple points and in independent lineages throughout the evolution of bony fishes. PMID- 12654891 TI - A second glutamine synthetase gene with expression in the gills of the gulf toadfish (Opsanus beta). AB - We characterized the expression of the nitrogen metabolism enzyme glutamine synthetase [GSase; L-glutamate: ammonia ligase (ADP-forming), E.C. 6.3.1.2] in tissues of the gulf toadfish Opsanus beta subjected to unconfined (ammonotelic) and confined (ureotelic) conditions. Enzymological results demonstrate that mass specific GSase activities rank in the order of brain > liver > stomach approximately kidney > intestine > gill > heart/spleen > muscle. When tissue mass is used to calculate a glutamine synthetic potential, the liver has the greatest, followed by muscle > stomach and intestine, with minor contributions from the remaining tissues. Additionally, during confinement stress, GSase activity increases significantly only in liver (fivefold) and muscle (twofold), tissues that previously showed significant expression of the other enzymes of urea synthesis. Western analyses of samples on SDS gels demonstrated that GSase specific protein content reflected enzyme activity, and all tissues except muscle had a single, similarly sized GSase subunit of 49.4 kDa; muscle showed staining of two bands of 36.8 and 98.9 kDa, which may possibly result from another gene product or post-translational modification. RT-PCR and RACE-PCR revealed the presence of a second GSase cDNA from gill tissue that shares only 73% nucleotide and amino acid sequence similarity with the GSase cDNA previously cloned from liver, and that lacks a mitochondrial leader-targeting sequence. RT-PCR and restriction digestion experiments demonstrated that mRNA from the original 'liver' GSase is expressed in all tissues examined (liver, gill, stomach, intestine, kidney, brain and muscle), whereas the new 'gill' form shows expression primarily in the gill. Gill GSase activity shows apparently exclusive expression in the soluble compartment, while other tissues expressing the 'liver' form show both cytoplasmic and mitochondrial activities. Phylogenetic analysis of a number of GSases demonstrates that the toadfish gill GSase has a greater affinity for a clade that includes the Xenopus GSase genes and one of two Fugu GSase genes, than it has for a clade containing the toadfish liver GSase and other described teleost GSase genes. The results are discussed in the context of a prior hypothesis on an ammonia-trapping mechanism in the gill of the toadfish. PMID- 12654892 TI - Twilight orientation to polarised light in the crepuscular dung beetle Scarabaeus zambesianus. AB - The polarisation pattern of skylight offers many arthropods a reference for visual compass orientation. The dung beetle Scarabaeus zambesianus starts foraging at around sunset. After locating a source of fresh droppings, it forms a ball of dung and rolls it off at high speed to escape competition at and around the dung pile. Using behavioural experiments in the field and in the laboratory, we show that the beetle is able to roll along a straight path by using the polarised light pattern of evening skylight. The receptors used to detect this skylight cue can be found in the ommatidia of the dorsal rim area of the eye, whose structures differ from the regular ommatidia in the rest of the eye. The dorsal rim ommatidia are characterised by rhabdoms with microvilli oriented at only two orthogonal orientations. Together with the finding that the receptors do not twist along the length of the rhabdom, this indicates that the photoreceptors of the dorsal rim area are polarisation sensitive. Large rhabdoms, a reflecting tracheal sheath and a lack of screening pigments make this area of the eye well adapted for polarised light detection at low light levels. The fan-shaped arrangement of receptors over the dorsal rim area was previously believed to be an adaptation to polarised light analysis, but here we argue that it is simply a consequence of the way that the eye is built. PMID- 12654893 TI - Functional consequences of a gene duplication and fusion event in an arginine kinase. AB - Arginine kinase (AK) from the foot of the razor clam Ensis directus consists of two full-length AK domains, denoted D1 and D2, fused in a single polypeptide chain. The full-length cDNA for Ensis AK was obtained and its deduced amino acid sequence was analyzed in the context of the X-ray crystal structure of a typical, monomeric AK. Both domains of Ensis AK contain most of the residues currently thought to be critical in catalysis, suggesting that both AK domains are catalytically competent. The full-length Ensis AK, a D2-NusA-His-tag fusion protein and a D2-truncated AK (enterokinase cleavage product of the fusion protein) were expressed in Escherichia coli and purified. All recombinant AK constructs displayed high enzyme activity. Attempts at expressing active D1 alone, D2 alone or a D1-NusA-His-tag fusion protein were unsuccessful. The catalytic properties of the active proteins were compared with the corresponding properties of recombinant AK from the horseshoe crab Limulus polyphemus, which is a typical monomeric AK. In contrast to expectations, the kinetic results strongly suggest that Ensis AK has only one active domain, namely D2. The K(cat) values for all Ensis constructs were roughly twice that of typical AKs, indicating higher overall catalytic throughput at the competent active site. Furthermore, both the full-length and truncated D2 Ensis AKs showed no synergism of substrate binding unlike typical AKs. The D2-NusA-His-tag fusion construct actually displayed negative synergism of substrate binding, which means that, in effect, the first substrate bound acts as a competitive inhibitor of the second. The conservation of the structure of the apparently inactive D1 may be related to constraints imposed by structural changes that could potentially impact substrate binding in D2 and/or possibly influence the proper folding of the enzyme during synthesis. Overall, the results from the present study indicate that the AK contiguous dimer from Ensis directus functions with activity in only the second domain. Although lacking activity in D1, D2 appears to compensate by having a higher intrinsic catalytic throughput than typical 40-kDa monomeric AKs. PMID- 12654894 TI - The energetics of the trot-gallop transition. AB - Two studies have focused on potential triggers for the trot-gallop transition in the horse. One study concluded that the transition was triggered by metabolic economy. The second study found that it was not metabolic factors but, rather, peak musculoskeletal forces that determine gait transition speeds. In theory, peak musculoskeletal forces should be the same when trotting up an incline as when trotting at the same speed on the level. Assuming this is the case, we hypothesized that if peak forces determine gait transition speeds then horses should switch from a trot to a gallop at the same speed (i.e. the same critical force) regardless of incline. The aim of the present research was to examine the effects of incline on the trot-gallop transition speed in horses and to re examine the role of metabolism in determining the trot-gallop transition. Horses (Equus caballus) were conditioned to run on a high-speed treadmill prior to data collection. Gait changes were recorded for each horse using a standardized testing protocol on the level and when trotting up a 10% incline. Both maximum sustained trotting speeds and minimum sustained galloping speeds, representing the lower and upper limits of the trot-gallop transition, respectively, were significantly slower when trotting up an incline. After completing collection of gait transition data, the horses were trained to extend their gaits beyond the normal transition speeds, and metabolic data were collected. Maximum sustained trotting speeds were not different from the energetically optimal transition speeds, i.e. the speed at which metabolic rates are the same for both gaits. PMID- 12654895 TI - Regulation of promoter occupancy during activation of cryptobiotic embryos from the crustacean Artemia franciscana. AB - Artemia franciscana embryos can suspend their development and metabolism at the gastrula stage to enter a state of cryptobiosis, forming cysts. Embryonic development and metabolism can be resumed under favorable environmental conditions to give rise to free-swimming larvae or nauplii. The mechanisms that mediate these processes are not completely known. Here, we report our studies of the mechanisms that regulate transcriptional activation upon exiting cryptobiosis. Regulatory regions of several A. franciscana gene promoters were identified. Functional analyses in mammalian cells allowed the identification of transcriptional activator regions in the Actin302 promoter and in promoter 2 of the sarco/endoplasmic reticulum Ca(2+)-ATPase-encoding gene. These regions were shown to specifically bind protein factors from nuclear extracts of A. franciscana nauplii by means of electrophoretic mobility shift assays. Several protein-binding regions were also detected by DNase I protection analysis in the promoters of the genes encoding the alpha1 subunit of Na(+)/K(+)-ATPase, actin 302 and sarco/endoplasmic reticulum Ca(2+)-ATPase. Specific DNA-binding proteins in nauplius nuclear extracts were detected for all the promoter regions analyzed. These proteins were either not present in cyst nuclear extracts or were present in much smaller concentrations. Three of the five regions analyzed also bound proteins present in cyst nuclear extracts. These data indicate that transcriptional activation upon exiting cryptobiosis in A. franciscana involves the expression/activation of DNA-binding transcription factors that are not present in cyst nuclei PMID- 12654896 TI - Adaptation in pheromone-sensitive trichoid sensilla of the hawkmoth Manduca sexta. AB - In extracellular tip recordings from long trichoid sensilla of male Manduca sexta moths, we studied dose-response relationships in response to bombykal stimuli of two different durations in the adapted and the non-adapted state. Bombykal responsive cells could be distinguished from non-bombykal-sensitive cells in each trichoid sensillum because the bombykal-responsive cell always generated the action potentials of larger initial amplitude. The bombykal cell, which was recorded at a defined location within a distal flagellar annulus, can resolve at least four log(10)-units of pheromone concentrations but is apparently unable to encode all stimulus durations tested. Parameters of the amplitude-modulated sensillar potential and the frequency-modulated action potential responses were examined in different states of adaptation. Evidence is presented for the existence of several mechanisms of adaptation, which affect distinct steps of the transduction cascade. After adapting pheromone stimuli, the sensillar potential rises to a lower amplitude and declines faster compared with the non-adapted response. In addition, the frequency of the adapted action potential response is reduced. Only the time of rise of the sensillar potential is differentially affected by adapting pheromone stimuli of different duration. The time of rise does not increase after short, but only after long, adapting stimuli. Both short and long adapting stimuli shift the dose-response curves of the sensillar potential amplitude, as well as the initial slope of its rising phase, to higher stimulus concentrations by approximately one log(10)-unit. The shift in the dose response curve of the action potential response is larger than for the sensillar potential response, suggesting that an additional adaptation mechanism acts at the level of action potential generation. Furthermore, a faster decline of the sensillar potential after short and long adapting stimuli suggests that the resting potential of the olfactory receptor neuron is stabilized. PMID- 12654897 TI - Regulation of L-alanine transport systems A and ASC by cyclic AMP and calcium in a reptilian duodenal model. AB - The regulation of neutral amino acid transport by cyclic AMP (cAMP) and calcium across the isolated duodenum of the lizard Gallotia galloti has been studied under short-circuit conditions. Active L-alanine transport was stimulated by forskolin, theophylline and dibutyryl cyclic AMP (db-cAMP). All these agents increased transmural potential difference (PD) and short-circuit current (I(sc)) in a manner consistent with the activation of a chloride secretory pathway. Both forskolin and theophylline increased intracellular cAMP levels in the lizard duodenal mucosa. Addition of calcium ionophore A23187 rapidly reduced mucosa-to serosa L-alanine fluxes and diminished net L-alanine transport. Despite the reduction of alanine fluxes by A23187, transepithelial PD and I(sc) values were increased by the ionophore. Analyses of the responses of isolated transport pathways indicated that the Na(+)-independent L-alanine transport system was unaffected by db-cAMP or calcium ionophore. By contrast, Na(+)-dependent transport activities were profoundly modified by these agents. Thus, while system A [alpha-methylamino-isobutiric acid (MeAIB)-transporting pathway] was stimulated by increased calcium, system ASC activity was nearly abolished. Calcium ionophore also potentiated the electrogenic response of system A. Forskolin strongly stimulated system ASC activity but left system A activity unchanged. Activation of system ASC by forskolin was clearly electroneutral, as pre-incubation of the tissues with the chloride channel blocker diphenylamine-2-carboxilic acid (DPC) completely prevented forskolin-induced transepithelial electrical responses. It is concluded that intracellular messengers cAMP and calcium oppositely modulate active Na(+)-dependent (L)-alanine transport in the lizard intestine. The different sensitivity exhibited by individual transport pathways may well account for the changes observed in overall alanine transport. PMID- 12654898 TI - CH-ILKBP regulates cell survival by facilitating the membrane translocation of protein kinase B/Akt. AB - Cell survival depends on proper propagation of protective signals through intracellular signaling intermediates. We report here that calponin homology domain-containing integrin-linked kinase (ILK)-binding protein (CH-ILKBP), a widely expressed adaptor protein localized at plasma membrane-actin junctions, is essential for transmission of survival signals. Cells that are depleted of CH ILKBP undergo extensive apoptosis despite the presence of cell-extracellular matrix contacts and soluble growth factors. The activating phosphorylation of protein kinase B (PKB/Akt), a key regulator of apoptosis, is impaired in the absence of CH-ILKBP. Importantly, loss of CH-ILKBP prevents the membrane translocation of PKB/Akt. Furthermore, forced membrane targeting of PKB/Akt bypasses the requirement of CH-ILKBP for the activating phosphorylation of PKB/Akt, suggesting that CH-ILKBP is required for the membrane translocation but not the subsequent phosphorylation of PKB/Akt. Finally, we show that loss of CH ILKBP is also required for the full activation of extracellular signal-regulated kinase (ERK)1/2. However, restoration of the PKB/Akt activation is sufficient for protection of cells from apoptosis induced by the depletion of CH-ILKBP despite the persistent suppression of the ERK1/2 activation. Thus, CH-ILKBP is an important component of the prosurvival signaling pathway functioning primarily by facilitating the membrane translocation of PKB/Akt and consequently the activation of PKB/Akt in response to extracellular survival signals. PMID- 12654899 TI - Dictyostelium cell death: early emergence and demise of highly polarized paddle cells. AB - Cell death in the stalk of Dictyostelium discoideum, a prototypic vacuolar cell death, can be studied in vitro using cells differentiating as a monolayer. To identify early events, we examined potentially dying cells at a time when the classical signs of Dictyostelium cell death, such as heavy vacuolization and membrane lesions, were not yet apparent. We observed that most cells proceeded through a stereotyped series of differentiation stages, including the emergence of "paddle" cells showing high motility and strikingly marked subcellular compartmentalization with actin segregation. Paddle cell emergence and subsequent demise with paddle-to-round cell transition may be critical to the cell death process, as they were contemporary with irreversibility assessed through time lapse videos and clonogenicity tests. Paddle cell demise was not related to formation of the cellulose shell because cells where the cellulose-synthase gene had been inactivated underwent death indistinguishable from that of parental cells. A major subcellular alteration at the paddle-to-round cell transition was the disappearance of F-actin. The Dictyostelium vacuolar cell death pathway thus does not require cellulose synthesis and includes early actin rearrangements (F actin segregation, then depolymerization), contemporary with irreversibility, corresponding to the emergence and demise of highly polarized paddle cells. PMID- 12654900 TI - The Ulp1 SUMO isopeptidase: distinct domains required for viability, nuclear envelope localization, and substrate specificity. AB - Protein modification by the ubiquitin-like SUMO protein contributes to many cellular regulatory mechanisms. In Saccharomyces cerevisiae, both sumoylating and desumoylating activities are essential for viability. Of its two known desumoylating enzymes, Ubl-specific protease (Ulp)1 and Ulp2/Smt4, Ulp1 is specifically required for cell cycle progression. A approximately 200-residue segment, the Ulp domain (UD), is conserved among Ulps and includes a core cysteine protease domain that is even more widespread. Here we demonstrate that the Ulp1 UD by itself can support wild-type growth rates and in vitro can cleave SUMO from substrates. However, in cells expressing only the UD of Ulp1, many SUMO conjugates accumulate to high levels, indicating that the nonessential Ulp1 NH2 terminal domain is important for activity against a substantial fraction of sumoylated targets. The NH2-terminal domain also includes sequences necessary and sufficient to concentrate Ulp1 at nuclear envelope sites. Remarkably, NH2 terminally deleted Ulp1 variants are able, unlike full-length Ulp1, to suppress defects of cells lacking the divergent Ulp2 isopeptidase. Thus, the NH2-terminal regulatory domain of Ulp1 restricts Ulp1 activity toward certain sumoylated proteins while enabling the cleavage of others. These data define key functional elements of Ulp1 and strongly suggest that subcellular localization is a physiologically significant constraint on SUMO isopeptidase specificity. PMID- 12654901 TI - Mid2p stabilizes septin rings during cytokinesis in fission yeast. AB - Septins are filament-forming proteins with a conserved role in cytokinesis. In the fission yeast Schizosaccharomyces pombe, septin rings appear to be involved primarily in cell-cell separation, a late stage in cytokinesis. Here, we identified a protein Mid2p on the basis of its sequence similarity to S. pombe Mid1p, Saccharomyces cerevisiae Bud4p, and Candida albicans Int1p. Like septin mutants, mid2delta mutants had delays in cell-cell separation. mid2delta mutants were defective in septin organization but not contractile ring closure or septum formation. In wild-type cells, septins assembled first during mitosis in a single ring and during septation developed into double rings that did not contract. In mid2delta cells, septins initially assembled in a single ring but during septation appeared in the cleavage furrow, forming a washer or disc structure. FRAP studies showed that septins are stable in wild-type cells but exchange 30 fold more rapidly in mid2delta cells. Mid2p colocalized with septins and required septins for its localization. A COOH-terminal pleckstrin homology domain of Mid2p was required for its localization and function. No genetic interactions were found between mid2 and the related gene mid1. Thus, these studies identify a new factor responsible for the proper stability and function of septins during cytokinesis. PMID- 12654902 TI - Nuclear envelope breakdown in starfish oocytes proceeds by partial NPC disassembly followed by a rapidly spreading fenestration of nuclear membranes. AB - Breakdown of the nuclear envelope (NE) was analyzed in live starfish oocytes using a size series of fluorescently labeled dextrans, membrane dyes, and GFP tagged proteins of the nuclear pore complex (NPC) and the nuclear lamina. Permeabilization of the nucleus occurred in two sequential phases. In phase I the NE became increasingly permeable for molecules up to approximately 40 nm in diameter, concurrent with a loss of peripheral nuclear pore components over a time course of 10 min. The NE remained intact on the ultrastructural level during this time. In phase II the NE was completely permeabilized within 35 s. This rapid permeabilization spread as a wave from one epicenter on the animal half across the nuclear surface and allowed free diffusion of particles up to approximately 100 nm in diameter into the nucleus. While the lamina and nuclear membranes appeared intact at the light microscopic level, a fenestration of the NE was clearly visible by electron microscopy in phase II. We conclude that NE breakdown in starfish oocytes is triggered by slow sequential disassembly of the NPCs followed by a rapidly spreading fenestration of the NE caused by the removal of nuclear pores from nuclear membranes still attached to the lamina. PMID- 12654903 TI - Spindle assembly and cytokinesis in the absence of chromosomes during Drosophila male meiosis. AB - A large body of work indicates that chromosomes play a key role in the assembly of both a centrosomal and centrosome-containing spindles. In animal systems, the absence of chromosomes either prevents spindle formation or allows the assembly of a metaphase-like spindle that fails to evolve into an ana-telophase spindle. Here, we show that Drosophila secondary spermatocytes can assemble morphologically normal spindles in the absence of chromosomes. The Drosophila mutants fusolo and solofuso are severely defective in chromosome segregation and produce secondary spermatocytes that are devoid of chromosomes. The centrosomes of these anucleated cells form robust asters that give rise to bipolar spindles that undergo the same ana-telophase morphological transformations that characterize normal spindles. The cells containing chromosome-free spindles are also able to assemble regular cytokinetic structures and cleave normally. In addition, chromosome-free spindles normally accumulate the Aurora B kinase at their midzones. This suggests that the association of Aurora B with chromosomes is not a prerequisite for its accumulation at the central spindle, or for its function during cytokinesis. PMID- 12654905 TI - Covalent trimers of the internal N-terminal trimeric coiled-coil of gp41 and antibodies directed against them are potent inhibitors of HIV envelope-mediated cell fusion. AB - We have engineered two soluble, covalently linked, trimeric polypeptides, N35CCG N13 and N34CCG comprising only the internal trimeric coiled-coil of the ectodomain of HIV-1 gp41. Both trimers inhibit human immunodeficiency virus, type 1 (HIV-1) envelope (Env)-mediated cell fusion at nanomolar concentrations by targeting the exposed C-terminal region of the gp41 ectodomain in the prehairpin intermediate state. The IC50 values for N35CCG-N13 and N34CCG are approximately 15 and approximately 95 nM, respectively, in a quantitative vaccinia virus-based reporter gene assay for HIV-1 Env-mediated cell fusion using Env from the T cell tropic strain LAV. Polyclonal antibodies were raised against N35CCG-N13 and a tightly binding fraction of anti-N35CCG-N13 inhibits T cell and macrophage tropic HIV-1 Env-mediated cell fusion with respective IC50 values of approximately 0.5 and approximately 1.5 microg/ml at 37 degrees C. The tightly binding anti-N35CCG N13 antibody fraction targets the exposed internal trimeric coiled-coil in the prehairpin intermediate state of gp41 in a manner analogous to peptides derived from the C region of the gp41 ectodomain. The potency of the tightly binding anti N35CCG-N13 antibody fraction in the fusion assay is comparable with that of the broadly neutralizing monoclonal antibody 2G12. These results indicate that N35CCG N13 is a potential anti-HIV therapeutic agent and represents a suitable immunogen for the generation of neutralizing monoclonal antibodies targeted to the internal trimeric coiled-coil of gp41. The data on the tightly binding anti-N35CCG-N13 antibody fraction demonstrate that the internal trimeric coiled-coil of gp41 in the prehairpin intermediate state is accessible to antibodies and that access is not restricted by either antibody size or the presence of a kinetic barrier. PMID- 12654906 TI - Binding discrimination of MutS to a set of lesions and compound lesions (base damage and mismatch) reveals its potential role as a cisplatin-damaged DNA sensing protein. AB - The DNA mismatch repair (MMR) system plays a critical role in sensitizing both prokaryotic and eukaryotic cells to the clinically potent anticancer drug cisplatin. It is thought to mediate cytotoxicity through recognition of cisplatin DNA lesions. This drug generates a range of lesions that may also give rise to compound lesions resulting from the misincorporation of a base during translesion synthesis. Using gel mobility shift competition assays and surface plasmon resonance, we have analyzed the interaction of Escherichia coli MutS protein with site-specifically modified DNA oligonucleotides containing each of the four cisplatin cross-links or a set of compound lesions. The major 1,2-d(GpG) cisplatin intrastrand cross-link was recognized with only a 1.5-fold specificity, whereas a 47-fold specificity was found with a natural G/T containing DNA substrate. The rate of association, kon, for binding to the 1,2-d(GpG) adduct was 3.1 x 104 m-1 s-1 and the specificity of binding was essentially dependent on koff. DNA duplexes containing a single 1,2-d(ApG), 1,3-d(GpCpG) adduct, and an interstrand cross-link of cisplatin were not preferentially recognized. Among 12 DNA substrates, each containing a different cisplatin compound lesion derived from replicative misincorporation of one base opposite either of the 1,2 intrastrand adducts, 10 were specifically recognized including those that are more likely formed in vivo based on cisplatin mutation spectra. Moreover, among these lesions, two compound lesions formed when an adenine was misincorporated opposite a 1,2-d(GpG) adduct were not substrates for the MutY-dependent mismatch repair pathway. The ability of MutS to sense differentially various platinated DNA substrates suggests that cisplatin compound lesions formed during misincorporation of a base opposite either adducted base of both 1,2-intrastrand cross-links are more plausible critical lesions for MMR-mediated cisplatin cytotoxicity. PMID- 12654908 TI - The mechanism of regulation of bacteriophage lambda pR promoter activity by Escherichia coli DnaA protein. AB - Apart from its function as an initiator of DNA replication, the Escherichia coli DnaA protein is also a specific transcription factor. It activates and represses a number of promoters. However, mechanisms of transcription stimulation by DnaA remained unknown. Bacteriophage lambda pR promoter is one of the promoters activated by DnaA. It was reported previously that DnaA binds downstream of the pR promoter and perhaps interacts with the RNA polymerase beta subunit. Here we demonstrate that DnaA positively regulates transcription from pR by stimulation of two steps in transcription initiation: RNA polymerase binding to the promoter region and promoter escape. For this transcription activation, two weak DnaA boxes located downstream of pR are necessary and sufficient. Such a mechanism of transcription activation and location of the activator-binding sites relative to the transcription start point are unusual in prokaryotes. Changes in the distance between the transcription start point and the first DnaA box by 5 and 10 bp and alterations in the orientation of these boxes did not abolish the stimulation of transcription by DnaA, but the efficiency of the promoter activation was different for various mutations. It seems plausible that formation of higher order nucleoprotein structures, involving DNA looping, is necessary for effective stimulation of the pR promoter. At high concentrations, DnaA is a repressor of pR rather than an activator. This repression was found to be because of inhibition of RNA polymerase binding to the promoter region. PMID- 12654904 TI - The Ran GTPase cycle is required for yeast nuclear pore complex assembly. AB - Here, we report the first evidence that the Ran GTPase cycle is required for nuclear pore complex (NPC) assembly. Using a genetic approach, factors required for NPC assembly were identified in Saccharomyces cerevisiae. Four mutant complementation groups were characterized that correspond to respective mutations in genes encoding Ran (gsp1), and essential Ran regulatory factors Ran GTPase activating protein (rna1), Ran guanine nucleotide exchange factor (prp20), and the RanGDP import factor (ntf2). All the mutants showed temperature-dependent mislocalization of green fluorescence protein (GFP)-tagged nucleoporins (nups) and the pore-membrane protein Pom152. A decrease in GFP fluorescence associated with the nuclear envelope was observed along with an increase in the diffuse, cytoplasmic signal with GFP foci. The defects did not affect the stability of existing NPCs, and nup mislocalization was dependent on de novo protein synthesis and continued cell growth. Electron microscopy analysis revealed striking membrane perturbations and the accumulation of vesicles in arrested mutants. Using both biochemical fractionation and immunoelectron microscopy methods, these vesicles were shown to contain nups. We propose a model wherein a Ran-mediated vesicular fusion step is required for NPC assembly into intact nuclear envelopes. PMID- 12654909 TI - The merozoite surface protein 1 complex of human malaria parasite Plasmodium falciparum: interactions and arrangements of subunits. AB - The major protein component at the surface of merozoites, the infectious form of blood stage malaria parasites, is the merozoite surface protein 1 (MSP-1) complex. In the human malaria parasite Plasmodium falciparum, this complex is generated by proteolytic cleavage of a 190-kDa glycosylphosphatidylinositol anchored precursor into four major fragments, which remain non-covalently associated. Here, we describe the in vitro reconstitution of the MSP-1 complex of P. falciparum strain 3D7 from its heterologously produced subunits. We provide evidence for the arrangement of the subunits within the complex and show how they interact with each other. Our data indicate that the conformation assumed by the reassembled complex as well as by the heterologously produced 190-kDa precursor corresponds to the native one. Based on these results we propose a first structural model for the MSP-1 complex. Together with access to faithfully produced material, this information will advance further structure-function studies of MSP-1 that plays an essential role during invasion of erythrocytes by the parasite and that is considered a promising candidate for a malaria vaccine. PMID- 12654911 TI - A recombinant chimeric epidermal growth factor-like module with high binding affinity for integrins. AB - Integrins are cell surface receptors involved in numerous pathological processes such as metastasis invasion and abnormal angiogenesis. To target these receptors, the epidermal growth factor (EGF)-like domain of human complement protease C1r was used as a natural scaffold to design chimeric modules containing the RGD motif. Here we report a high yield bacterial expression system and its application to the production of two such modules, EGF-RGD and V2, the latter variant mimicking the RGD-containing domain of disintegrins. These modules were characterized chemically, and their biological activity was investigated by cellular assays using various Chinese hamster ovary cell lines expressing beta1 and beta3 integrins and by surface plasmon resonance spectroscopy. Remarkably, the modifications leading to the V2 variant had differential effects on the interaction with beta3 and beta1 integrins. The disintegrin-like V2 module exhibited enhanced binding affinities compared with EGF-RGD, with KD values of 7.2 nm for alpha5beta1 (a 4-fold decrease) and 3.5 nm for alphavbeta3 (a 1.5-fold decrease), comparable with the values determined for natural integrin ligands. Analysis by NMR spectroscopy also revealed a differential dynamic behavior of the RGD motif in the EGF-RGD and V2 variants, providing insights into the structural basis of their relative binding efficiency. These novel RGD-containing EGF modules open the way to the design of improved variants with selective affinity for particular integrins and their use as carriers for other biologically active modules. PMID- 12654910 TI - The alpha-glucuronidase, GlcA67A, of Cellvibrio japonicus utilizes the carboxylate and methyl groups of aldobiouronic acid as important substrate recognition determinants. AB - alpha-Glucuronidases are key components of the ensemble of enzymes that degrade the plant cell wall. They hydrolyze the alpha1,2-glycosidic bond between 4-O methyl-d-glucuronic acid (4-O-MeGlcA) and the xylan or xylooligosaccharide backbone. Here we report the crystal structure of an inactive mutant (E292A) of the alpha-glucuronidase, GlcA67A, from Cellvibrio japonicus in complex with its substrate. The data show that the 4-O-methyl group of the substrate is accommodated within a hydrophobic sheath flanked by Val-210 and Trp-160, whereas the carboxylate moiety is located within a positively charged region of the substrate-binding pocket. The carboxylate side chains of Glu-393 and Asp-365, on the "beta-face" of 4-O-MeGlcA, form hydrogen bonds with a water molecule that is perfectly positioned to mount a nucleophilic attack at the anomeric carbon of the target glycosidic bond, providing further support for the view that, singly or together, these amino acids function as the catalytic base. The capacity of reaction products and product analogues to inhibit GlcA67A shows that the 4-O methyl group, the carboxylate, and the xylose sugar of aldobiouronic acid all play an important role in substrate binding. Site-directed mutagenesis informed by the crystal structure of enzyme-ligand complexes was used to probe the importance of highly conserved residues at the active site of GlcA67A. The biochemical properties of K288A, R325A, and K360A show that a constellation of three basic amino acids (Lys-288, Arg-325, and Lys-360) plays a critical role in binding the carboxylate moiety of 4-O-MeGlcA. Disruption of the apolar nature of the pocket created by Val-210 (V210N and V210S) has a detrimental effect on substrate binding, although the reduction in affinity is not reflected by an inability to accommodate the 4-O-methyl group. Replacing the two tryptophan residues that stack against the sugar rings of the substrate with alanine (W160A and W543A) greatly reduced activity. PMID- 12654913 TI - Lectins homologous to those of monocotyledonous plants in the skin mucus and intestine of pufferfish, Fugu rubripes. AB - We have characterized pufflectin, a novel mannose-specific lectin, from the skin mucus of the pufferfish, Fugu rubripes. Molecular mass estimations by gel filtration and matrix-assisted laser desorption ionization time-of-flight mass spectrometry and the SDS-PAGE pattern suggest that pufflectin is a homodimer composed of non-covalently associated subunits of 13 kDa. The full-length pufflectin cDNA consists of 527 bp, with 116 amino acid residues deduced from the open reading frame. The amino acid sequence of pufflectin shows no homology with any known animal lectin. Surprisingly, pufflectin shares sequence homology with mannose-binding lectins of monocotyledonous plants and has conserved two of three carbohydrate recognition domains of these plant lectins. The pufflectin gene is expressed in gills, oral cavity wall, esophagus, and skin. In addition, an isoform occurs exclusively in the intestine. Pufflectin differs from mannose binding lectins purified from the blood plasma of Fugu. Whereas pufflectin did not agglutinate five bacterial species tested, it was demonstrated to bind to the parasitic trematode, Heterobothrium okamotoi. This finding suggests that pufflectin contributes to the parasite-defense system in Fugu. PMID- 12654912 TI - Vps9p CUE domain ubiquitin binding is required for efficient endocytic protein traffic. AB - Rab5 GTPases are key regulators of protein trafficking through the early stages of the endocytic pathway. The yeast Rab5 ortholog Vps21p is activated by its guanine nucleotide exchange factor Vps9p. Here we show that Vps9p binds ubiquitin and that the CUE domain is necessary and sufficient for this interaction. Vps9p ubiquitin binding is required for efficient endocytosis of Ste3p but not for the delivery of the biosynthetic cargo carboxypeptidase Y to the vacuole. In addition, Vps9p is itself monoubiquitylated. Ubiquitylation is dependent on a functional CUE domain and Rsp5p, an E3 ligase that participates in cell surface receptor endocytosis. These findings define a new ubiquitin binding domain and implicate ubiquitin as a modulator of Vps9p function in the endocytic pathway. PMID- 12654914 TI - A novel zinc snap motif conveys structural stability to 3-methyladenine DNA glycosylase I. AB - The Escherichia coli 3-methyladenine DNA glycosylase I (TAG) is a DNA repair enzyme that excises 3-methyladenine in DNA and is the smallest member of the helix-hairpin-helix (HhH) superfamily of DNA glycosylases. Despite many studies over the last 25 years, there has been no suggestion that TAG was a metalloprotein. However, here we establish by heteronuclear NMR and other spectroscopic methods that TAG binds 1 eq of Zn2+ extremely tightly. A family of refined NMR structures shows that 4 conserved residues contributed from the amino and carboxyl-terminal regions of TAG (Cys4, His17, His175, and Cys179) form a Zn2+ binding site. The Zn2+ ion serves to tether the otherwise unstructured amino and carboxyl-terminal regions of TAG. We propose that this unexpected "zinc snap" motif in the TAG family (CX(12-17)HX(approximately 150)HX(3)C) serves to stabilize the HhH domain thereby mimicking the functional role of protein-protein interactions in larger HhH superfamily members. PMID- 12654915 TI - Mechanism of the targeting action of DnaJ in the DnaK molecular chaperone system. AB - In the DnaK (Hsp70) molecular chaperone system of Escherichia coli, the substrate polypeptide is fed into the chaperone cycle by association with the fast-binding, ATP-liganded form of the DnaK. The substrate binding properties of DnaK are controlled by its two cochaperones DnaJ (Hsp40) and GrpE. DnaJ stimulates the hydrolysis of DnaK-bound ATP, and GrpE accelerates ADP/ATP exchange. DnaJ has been described as targeting the substrate to DnaK, a concept that has remained rather obscure. Based on binding experiments with peptides and polypeptides we propose here a novel mechanism for the targeting action of DnaJ: ATP.DnaK and DnaJ with its substrate-binding domain bind to different segments of one and the same polypeptide chain forming (ATP.DnaK)m.substrate.DnaJn complexes; in these ternary complexes efficient cis-interaction of the J-domain of DnaJ with DnaK is favored by their propinquity and triggers the hydrolysis of DnaK-bound ATP, converting DnaK to its ADP-liganded high affinity state and thus locking it onto the substrate polypeptide. PMID- 12654916 TI - Questioning the role of checkpoint kinase 2 in the p53 DNA damage response. AB - Cdc25C and p53 have been reported to be physiological targets of checkpoint kinase 2 (Chk2). Surprisingly, although Chk2 purified from DNA damage sustaining cells has dramatically increased ability to phosphorylate Cdc25C when compared with untreated cells, its ability to phosphorylate p53 is weak before treatment, and there is no increase in its activity toward p53 after DNA damage by gamma irradiation or the radiomimetic agent neocarzinostatin. Furthermore, introduction of Chk2 short interfering RNA into three different human tumor cell lines leads to marked reduction of Chk2 protein, but p53 is still stabilized and active after DNA damage. The results with Chk1 short interfering RNA indicate as well that Chk1 does not play a role in human p53 stabilization after DNA damage. Thus, Chk1 and Chk2 are unlikely to be regulators of p53 in at least some human tumor cells. We discuss our results in the context of previous findings demonstrating a requirement for Chk2 in p53 stabilization and activity. PMID- 12654917 TI - The Chk2 tumor suppressor is not required for p53 responses in human cancer cells. AB - Ionizing radiation damages chromosomal DNA and activates p53-dependent transcription in mammalian cells. The Chk2 protein kinase has been hypothesized to be the primary mediator of this response. We have rigorously tested this hypothesis in human cells by disrupting the CHK2 gene through homologous recombination. We found that the p53 response was unexpectedly robust in such cells. Phosphorylation of p53 at serine 20, accumulation of p53 protein, transcriptional activation of p53 target genes, and cell cycle arrest and apoptotic death phenotypes were completely intact regardless of CHK2 status. Our results indicate that Chk2 kinase is not required for p53 activation in human cells and explain why CHK2 and TP53 mutations can jointly occur in human tumors. PMID- 12654918 TI - Serine phosphorylation negatively regulates RhoA in vivo. AB - Previous work indicates that RhoA phosphorylation on Ser188 by cAMP or cGMP dependent kinases inhibits its activity. However, these studies lacked the possibility to directly study phosphorylated RhoA activity in vivo. Therefore, we created RhoA proteins containing phosphomimetic residues in place of the cAMP/cGMP-dependent kinase phosphorylation site. RhoA phosphorylation or phosphomimetic substitution did not affect Rho guanine nucleotide exchange factor, GTPase activating protein, or geranylgeranyl transferase activity in vitro but promoted binding to the Rho guanine-dissociation inhibitor as measured by exchange factor competition assays. The in vitro similarities between RhoA phosphomimetic proteins and phosphorylated RhoA allowed us to study function of phosphorylated RhoA in vivo. RhoA phosphomimetic proteins display depressed GTP loading when transiently expressed in NIH 3T3 cells. Stable-expressing RhoA and RhoA(S188A) clones spread significantly slower than mock-transfected or RhoA(S188E) clones. RhoA(S188A) clones were protected from the morphological effects of a cAMP agonist, whereas phosphomimetic clones exhibit stress fiber disassembly similar to control cells. Together, these data provide in vivo evidence that addition of a charged group to Ser188 upon phosphorylation negatively regulates RhoA activity and indicates that this occurs through enhanced Rho guanine-dissociation inhibitor interaction rather than direct perturbation of guanine nucleotide exchange factor, GTPase activating protein, or geranylgeranyl transferase activity. PMID- 12654919 TI - Subunit interaction and regulation of activity through terminal domains of the family D DNA polymerase from Pyrococcus horikoshii. AB - Functions of the terminal domains of the family D DNA polymerase from Pyrococcus horikoshii (PolDPho) were analyzed by making and characterizing various truncated proteins. Based on a co-expression vector developed previously (Shen, Y., Musti, K., Hiramoto, M., Kikuchi, H., Kawabayashi, Y., and Matsui, I. (2001) J. Biol. Chem. 276, 27376-27383), 25 vectors for terminal truncated proteins were constructed. The expressed proteins were characterized in terms of thermostability, subunit interaction, and polymerization and 3'-5' exonuclease activities. The carboxyl-terminal (1255-1332) of the large subunit (DP2Pho) and two regions, the 201-260 and 599-622, of the small subunit (DP1Pho) were found to be critical for the complex formation, and probable subunit interaction of PolDPho. The amino-terminal (1-300) of DP2Pho is essential for the folding of PolDPho and is likely the oligomerization domain of PolDPho. A short region at the extreme C-terminal of DP2Pho (from 1385 to 1434) and the N-terminal of DP1Pho(1-200), which forms a stable protein, are not absolutely necessary for either polymerization or the 3'-5' exonuclease activity. We identified a possible regulatory role of DP1Pho(1-200) for the 3'-5' exonuclease. Deletion of DP1Pho(1 200) increased the exonuclease and DNA binding activities of PolDPho. Adding DP1Pho(1-200) to the truncated protein suppressed the elevated exonuclease activity. We also constructed and analyzed three internal deletion mutants and two site-directed mutants in the region of the putative zinc finger motif (cysteine cluster II) of DP2Pho at the COOH-terminal. We found that the internal region of the zinc finger motif is critical for the 3'-5' exonuclease, but is dispensable for the DNA polymerization. PMID- 12654920 TI - Interactions between Piccolo and the actin/dynamin-binding protein Abp1 link vesicle endocytosis to presynaptic active zones. AB - Piccolo is a high molecular weight multi-domain protein shown to be a structural component of the presynaptic CAZ (cytoskeletal matrix assembled at active zones). These features indicate that Piccolo may act to scaffold proteins involved in synaptic vesicle endo- and exocytosis near their site of action. To test this hypothesis, we have utilized a functional cell-based endocytosis assay and identified the N-terminal proline-rich Q domain in Piccolo as a region that interferes with clathrin-mediated endocytosis. Utilizing the Piccolo Q domain as bait in a yeast two-hybrid screen, we have identified the F-actin-binding protein Abp1 (also called SH3P7 or HIP-55) as a potential binding partner for this domain. The physiological relevance of this interaction is supported by in vitro binding studies, colocalization in nerve terminals, in vivo recruitment studies, and immunoprecipitation experiments. Intriguingly, Abp1 binds to both F-actin and the GTPase dynamin and has been implicated in linking the actin cytoskeleton to clathrin-mediated endocytosis. Our results suggest that Piccolo, as a structural protein of the CAZ, may serve to localize Abp1 at active zones where it can actively participate in creating a functional connection between the dynamic actin cytoskeleton and synaptic vesicle recycling. PMID- 12654921 TI - Characterization of 2-enoyl thioester reductase from mammals. An ortholog of YBR026p/MRF1'p of the yeast mitochondrial fatty acid synthesis type II. AB - A data base search with YBR026c/MRF1', which encodes trans-2-enoyl thioester reductase of the intramitochondrial fatty acid synthesis (FAS) type II in yeast (Torkko, J. M., Koivuranta, K. T., Miinalainen, I. J., Yagi, A. I., Schmitz, W., Kastaniotis, A. J., Airenne, T. T., Gurvitz, A., and Hiltunen, K. J. (2001) Mol. Cell. Biol. 21, 6243-6253), revealed the clone AA393871 (HsNrbf-1, nuclear receptor binding factor 1) in human EST data bank. Expression of HsNrbf-1, tagged C-terminally with green fluorescent protein, in HeLa cells, resulted in a punctated fluorescence signal, superimposable with the MitoTracker Red dye. Wild type polypeptide was immunoisolated from the extract of bovine heart mitochondria. Recombinant HsNrbf-1p reduces trans-2-enoyl-CoA to acyl-CoA with chain length from C6 to C16 in an NADPH-dependent manner with preference to medium chain length substrate. Furthermore, expression of HsNRBF-1 in the ybr026cDelta yeast strain restored mitochondrial respiratory function allowing growth on glycerol. These findings provide evidence that Nrbf-1ps act as a mitochondrial 2-enoyl thioester reductase, and mammalian cells may possess bacterial type fatty acid synthetase (FAS type II) in mitochondria, in addition to FAS type I in the cytoplasm. PMID- 12654922 TI - Inhibition of InhA activity, but not KasA activity, induces formation of a KasA containing complex in mycobacteria. AB - Isoniazid (INH) remains one of the key drugs used to control tuberculosis, with the enoyl-AcpM reductase InhA being the primary target. However, based on the observation that INH-treated Mycobacterium tuberculosis overproduces KasA, an enzyme involved in the biosynthesis of mycolic acids, and induces the formation of a covalent complex consisting of AcpM, KasA, and INH, it has been proposed that KasA represents the primary target of INH. However, the relevance of this complex to INH action remains obscure. This study was aimed at clarifying the role of InhA and KasA in relation to INH activity. By using anti-KasA antibodies we detected the KasA-containing complex in INH-treated Mycobacterium smegmatis. In addition, INH-treated cells also produced constant levels of KasA that were not sequestered in the complex and presumably were sufficient to ensure mycolic acid biosynthesis. Interestingly, a furA-lacking strain induced the complex at lower concentrations of INH compared with the control strain, whereas higher INH concentrations were necessary to induce the complex in a strain that lacks katG, suggesting that INH needs to be activated by KatG to induce the KasA-containing complex. The InhA inhibitors ethionamide and diazaborine also induced the complex; thus, its formation was not specifically relevant to INH action but was because of InhA inhibition. In addition, in vitro assays using purified InhA and KasA demonstrated that KatG-activated INH, triclosan, and diazaborine inhibited InhA but not KasA activity. Moreover, several thermosensitive InhA mutant strains of M. smegmatis constitutively expressed the KasA-containing complex. This study provides the biochemical and genetic evidence. 1) Only inhibition of InhA, but not KasA, induces the KasA-containing complex. 2) INH is not part of the complex. 3) INH does not target KasA, consistent with InhA being the primary target of INH. PMID- 12654924 TI - Determinants of inhibition of transiently expressed voltage-gated calcium channels by omega-conotoxins GVIA and MVIIA. AB - The Conus magus peptide toxin omega-conotoxin MVIIA is considered an irreversible, specific blocker of N-type calcium channels, and is now in clinical trials as an intrathecal analgesic. Here, we have examined the action of MVIIA on mutant and wild type calcium channels transiently expressed in tsA-201 cells. Although we have shown previously that mutations in a putative external EF-hand motif in the domain IIIS5-H5 region alters block by both omega-conotoxin GVIA and MVIIA (Feng, Z. P., Hamid, J., Doering, C., Bosey, G. M., Snutch, T. P., and Zamponi, G. W. (2001) J. Biol. Chem. 276, 15728-15735), the introduction of five point mutations known to affect GVIA blocking (and located downstream of the EF hand) affected MVIIA block to a smaller degree compared with GVIA. These data suggest that despite some overlap, MVIIA and GVIA block does not share identical channel structural determinants. At higher concentrations (approximately 3 microm), MVIIA reversibly blocked L-, P/Q-, and R-type, but not T-type channels, indicating that the overall architecture of the MVIIA site is conserved in all types of high voltage-activated calcium channels. A kinetic analysis of the MVIIA effects on the N-type channel showed that MVIIA blocked resting, open, and inactivated channels. Although the development of MVIIA block did not appear to be voltage-, nor frequency-dependent, the degree of recovery from block strongly depended on the potential applied during washout. Interestingly, the degree of washout was highly variable and appeared to weakly depend on the holding potential applied during toxin application. We propose a model in which N-type calcium channels can form both reversible and irreversible complexes with MVIIA. PMID- 12654925 TI - Alteration of polysaccharide size distribution of a vertebrate hyaluronan synthase by mutation. AB - Hyaluronan (HA) is a nonsulfated glycosaminoglycan that has long been known to play structural roles in vertebrates. Recently, it has become increasingly obvious that this linear polysaccharide has many more uses than simply scaffolding or space filler. HA has been found to be involved in development, cell signaling, cell motility, and metastasis. These roles are often dictated by the length of the HA polymer, which can vary from a few to about 10,000 sugar residues in length. Three distinct isoforms of HA synthase exist in mammals. It has been shown previously by others that each isoform produces HA that differs in size distribution, but the regulatory mechanism is not yet known. Mutations have been described that alter the size distribution of the HA produced by the streptococcal HA synthases. We show that by mutating one particular amino acid residue of a vertebrate HA synthase, depending on the introduced side chain, the size of HA produced can be either reduced or increased. We postulate that several cysteine residues and a serine residue may be involved in binding directly or indirectly to the nascent HA chain. These data support the theory that the relative strength of the interaction between the catalyst and the polymer may be a major factor in HA size control. PMID- 12654926 TI - Regulation of interleukin-1alpha expression by integrins and epidermal growth factor receptor in keratinocytes from a mouse model of inflammatory skin disease. AB - Transgenic mice expressing beta1 integrins in the suprabasal epidermal layers have sporadic skin hyperproliferation and inflammation correlated with activation of extracellular signal-regulated kinase (Erk) mitogen-activated protein kinase and increased interleukin (IL)-1alpha production. We investigated the link between aberrant integrin expression, Erk activation, and expression of IL 1alpha. Transgenic keratinocytes had higher basal Erk activity and IL-1alpha levels than nontransgenic controls and were more sensitive to stimulation of Erk activity and IL-1alpha production by IL-1alpha, 12-O-tetradecanoylphorbol-13 acetate (TPA), epidermal growth factor (EGF), and serum. Inhibition of Erk in transgenic keratinocytes reduced basal IL-1alpha levels and the stimulation of IL 1alpha production by serum or phorbol ester, demonstrating that Erk could regulate IL-1alpha expression. TPA or IL-1alpha treatment resulted in rapid down regulation of the EGF receptor in transgenic cells, indicative of transactivation. Inhibition of transactivation blocked basal and TPA or IL-1alpha induced Erk activation, but not IkappaBalpha degradation, and abolished increased IL-1alpha production in transgenic cells. In transgene-negative cells, constitutive activation of IL-1-dependent signaling by wild type or kinase-dead IRAK1 stimulated IL-1alpha production independent of Erk. We conclude that suprabasal integrin expression leads to Erk activation and increased IL-1alpha expression by potentiating activation of the EGF receptor. These results provide a mechanism by which aberrant integrin expression triggers epidermal hyperproliferation and inflammation. PMID- 12654927 TI - Affinity and specificity of interactions between Nedd4 isoforms and the epithelial Na+ channel. AB - The epithelial Na+ channel (alphabetagammaENaC) regulates salt and fluid homeostasis and blood pressure. Each ENaC subunit contains a PY motif (PPXY) that binds to the WW domains of Nedd4, a Hect family ubiquitin ligase containing 3-4 WW domains and usually a C2 domain. It has been proposed that Nedd4-2, but not Nedd4-1, isoforms can bind to and suppress ENaC activity. Here we challenge this notion and show that, instead, the presence of a unique WW domain (WW3*) in either Nedd4-2 or Nedd4-1 determines high affinity interactions and the ability to suppress ENaC. WW3* from either Nedd4-2 or Nedd4-1 binds ENaC-PY motifs equally well (e.g. Kd approximately 10 microm for alpha- or betaENaC, 3-6-fold higher affinity than WW4), as determined by intrinsic tryptophan fluorescence. Moreover, dNedd4-1, which naturally contains a WW3* instead of WW2, is able to suppress ENaC function equally well as Nedd4-2. Homology models of the WW3*.betaENaC-PY complex revealed that a Pro and Ala conserved in all WW3*, but not other Nedd4-WW domains, help form the binding pocket for PY motif prolines. Extensive contacts are formed between the betaENaC-PY motif and the Pro in WW3*, and the small Ala creates a large pocket to accommodate the peptide. Indeed, mutating the conserved Pro and Ala in WW3* reduces binding affinity 2-3-fold. Additionally, we demonstrate that mutations in PY motif residues that form contacts with the WW domain based on our previously solved structure either abolish or severely reduce binding affinity to the WW domain and that the extent of binding correlates with the level of ENaC suppression. Independently, we show that a peptide encompassing the PY motif of sgk1, previously proposed to bind to Nedd4-2 and alter its ability to regulate ENaC, does not bind (or binds poorly) the WW domains of Nedd4-2. Collectively, these results suggest that high affinity of WW domain-PY-motif interactions rather than affiliation with Nedd4-1/Nedd-2 is critical for ENaC suppression by Nedd4 proteins. PMID- 12654929 TI - G+C3 structuring along the genome: a common feature in prokaryotes. AB - The heterogeneity of gene nucleotide content in prokaryotic genomes is commonly interpreted as the result of three main phenomena: (1) genes undergo different selection pressures both during and after translation (affecting codon and amino acid choice); (2) genes undergo different mutational pressure whether they are on the leading or lagging strand; and (3) genes may have different phylogenetic origins as a result of lateral transfers. However, this view neglects the necessity of organizing genetic information on a chromosome that needs to be replicated and folded, which may add constraints to single gene evolution. As a consequence, genes are potentially subjected to different mutation and selection pressures, depending on their position in the genome. In this paper, we analyze the structuring of different codon usage measures along completely sequenced bacterial genomes. We show that most of them are highly structured, suggesting that genes have different base content, depending on their location on the chromosome. A peculiar pattern of genome structure, with a tendency toward an A+T enrichment near the replication terminus, is found in most bacterial phyla and may reflect common chromosome constraints. Several species may have lost this pattern, probably because of genome rearrangements or integration of foreign DNA. We show that in several species, this enrichment is associated with an increase of evolutionary rate and we discuss the evolutionary implications of these results. We argue that structural constraints acting on the circular chromosome are not negligible and that this natural structuring of bacterial genomes may be a cause of overestimation in lateral gene transfer predictions using codon composition indices. PMID- 12654928 TI - Comparative analysis of pyruvate kinases from the hyperthermophilic archaea Archaeoglobus fulgidus, Aeropyrum pernix, and Pyrobaculum aerophilum and the hyperthermophilic bacterium Thermotoga maritima: unusual regulatory properties in hyperthermophilic archaea. AB - Pyruvate kinases (PK, EC 2.7.1.40) from three hyperthermophilic archaea (Archaeoglobus fulgidus strain 7324, Aeropyrum pernix, and Pyrobaculum aerophilum) and from the hyperthermophilic bacterium Thermotoga maritima were compared with respect to their thermophilic, kinetic, and regulatory properties. PKs from the archaea are 200-kDa homotetramers composed of 50-kDa subunits. The enzymes required divalent cations, Mg2+ and Mn2+ being most effective, but were independent of K+. Temperature optima for activity were 85 degrees C (A. fulgidus) and above 98 degrees C (A. pernix and P. aerophilum). The PKs were highly thermostable up to 110 degrees C (A. pernix) and showed melting temperatures for thermal unfolding at 93 degrees C (A. fulgidus) or above 98 degrees C (A. pernix and P. aerophilum). All archaeal PKs exhibited sigmoidal saturation kinetics with phosphoenolpyruvate (PEP) and ADP indicating positive homotropic cooperative response with both substrates. Classic heterotropic allosteric regulators of PKs from eukarya and bacteria, e.g. fructose 1,6 bisphosphate or AMP, did not affect PK activity of hyperthermophilic archaea, suggesting the absence of heterotropic allosteric regulation. PK from the bacterium T. maritima is also a homotetramer of 50-kDa subunits. The enzyme was independent of K+ ions, had a temperature optimum of 80 degrees C, was highly thermostable up to 90 degrees C, and had a melting temperature above 98 degrees C. The enzyme showed cooperative response to PEP and ADP. In contrast to its archaeal counterparts, the T. maritima enzyme exhibited the classic allosteric response to the activator AMP and to the inhibitor ATP. Sequences of hyperthermophilic PKs showed significant similarity to characterized PKs from bacteria and eukarya. Phylogenetic analysis of PK sequences of all three domains indicates a distinct archaeal cluster that includes the PK from the hyperthermophilic bacterium T. maritima. PMID- 12654930 TI - Influence of mutational and sampling factors on the estimation of demographic parameters in a "continuous" population under isolation by distance. AB - In numerous species, individual dispersal is restricted in space so that "continuous" populations evolve under isolation by distance. A method based on individual genotypes assuming a lattice population model was recently developed to estimate the product Dsigma2, where D is the population density and sigma2 is the average squared parent-offspring distance. We evaluated the influence on this method of both mutation rate and mutation model, with a particular reference to microsatellite markers, as well as that of the spatial scale of sampling. Moreover, we developed and tested a nonparametric bootstrap procedure allowing the construction of confidence intervals for the estimation of Dsigma2. These two objectives prompted us to develop a computer simulation algorithm based on the coalescent theory giving individual genotypes for a continuous population under isolation by distance. Our results show that the characteristics of mutational processes at microsatellite loci, namely the allele size homoplasy generated by stepwise mutations, constraints on allele size, and change of slippage rate with repeat number, have little influence on the estimation of Dsigma2. In contrast, a high genetic diversity (approximately 0.7-0.8), as is commonly observed for microsatellite markers, substantially increases the precision of the estimation. However, very high levels of genetic diversity (>0.85) were found to bias the estimation. We also show that statistics taking into account allele size differences give unreliable estimations (i.e., high variance of Dsigma2 estimation) even under a strict stepwise mutation model. Finally, although we show that this method is reasonably robust with respect to the sampling scale, sampling individuals at a local geographical scale gives more precise estimations of Dsigma2. PMID- 12654931 TI - Colonization of heterochromatic genes by transposable elements in Drosophila. AB - As a further step toward understanding transposable element-host genome interactions, we investigated the molecular anatomy of introns from five heterochromatic and 22 euchromatic protein-coding genes of Drosophila melanogaster. A total of 79 kb of intronic sequences from heterochromatic genes and 355 kb of intronic sequences from euchromatic genes have been used in Blast searches against Drosophila transposable elements (TEs). The results show that TE homologous sequences belonging to 19 different families represent about 50% of intronic DNA from heterochromatic genes. In contrast, only 0.1% of the euchromatic intron DNA exhibits homology to known TEs. Intraspecific and interspecific size polymorphisms of introns were found, which are likely to be associated with changes in TE-related sequences. Together, the enrichment in TEs and the apparent dynamic state of heterochromatic introns suggest that TEs contribute significantly to the evolution of genes located in heterochromatin. PMID- 12654932 TI - Ancient SINEs from African endemic mammals. AB - Afrotheria is a newly recognized taxon comprising elephants, hyraxes, sea cows, aardvarks, golden moles, tenrecs, and elephant shrews, each of which originated in Africa. Although some members of this taxon were once classified into distantly related groups, recent molecular studies have demonstrated their close relationships. It was suggested that this group emerged as a result of physical isolation of the African continent during the successive breakup events of Gondowanaland. In this study, a novel family of SINEs, designated AfroSINEs, was isolated and characterized from the genomes of afrotherians. This SINE family is distributed exclusively among the afrotherian species, confirming their monophyletic relationships. Furthermore, a distinct subfamily, which shares a deletion in the middle region of the SINE, was identified. The distribution of this subfamily is apparently restricted to the genomes of hyraxes, elephants, and sea cows, suggesting monophyly of these three groups, which was previously proposed as Paenungulata. We characterized the structures of the AfroSINEs from all afrotherian representatives by PCR, and we discuss how they were generated as well as the phylogenetic relationships of their host species. PMID- 12654933 TI - Rapid divergence of the ecdysone receptor in Diptera and Lepidoptera suggests coevolution between ECR and USP-RXR. AB - Ecdysteroid hormones are major regulators in reproduction and development of insects, including larval molts and metamorphosis. The functional ecdysone receptor is a heterodimer of ECR (NR1H1) and USP-RXR (NR2B4), which is the orthologue of vertebrate retinoid X receptors (RXR alpha, beta, gamma). Both proteins belong to the superfamily of nuclear hormone receptors, ligand-dependent transcription factors that share two conserved domains: the DNA-binding domain (DBD) and the ligand-binding domain (LBD). In order to gain further insight into the evolution of metamorphosis and gene regulation by ecdysone in arthropods, we performed a phylogenetic analysis of both partners of the heterodimer ECR/USP RXR. Overall, 38 USP-RXR and 19 ECR protein sequences, from 33 species, have been used for this analysis. Interestingly, sequence alignments and structural comparisons reveal high divergence rates, for both ECR and USP-RXR, specifically among Diptera and Lepidoptera. The most impressive differences affect the ligand binding domain of USP-RXR. In addition, ECR sequences show variability in other domains, namely the DNA-binding and the carboxy-terminal F domains. Our data provide the first evidence that ECR and USP-RXR may have coevolved during holometabolous insect diversification, leading to a functional divergence of the ecdysone receptor. These results have general implications on fundamental aspects of insect development, evolution of nuclear receptors, and the design of specific insecticides. PMID- 12654934 TI - Formation of solo-LTRs through unequal homologous recombination counterbalances amplifications of LTR retrotransposons in rice Oryza sativa L. AB - We studied the dynamics of hopi, Retrosat1, and RIRE3, three gypsy-like long terminal repeat (LTR) retrotransposons, in Oryza sativa L. genome. For each family, we assessed the phenetic relationships of the copies and estimated the date of insertion of the complete copies through the evaluation of their LTR divergence. We show that within each family, distinct phenetic groups have inserted at significantly different times, within the past 5 Myr and that two major amplification events may have occurred during this period. We show that solo-LTR formation through homologous unequal recombination has occurred in rice within the past 5 Myr for the three elements. We thus propose an increase/decrease model for rice genome evolution, in which both amplification and recombination processes drive variations in genome size. PMID- 12654935 TI - Significantly different patterns of amino acid replacement after gene duplication as compared to after speciation. AB - We have performed a large-scale analysis of amino acid sequence evolution after gene duplication by comparing evolution after gene duplication with evolution after speciation in over 1,800 phylogenetic trees constructed from manually curated alignments of protein domains downloaded from the PFAM database. The site specific rate of evolution is significantly altered by gene duplication. A significant increase in the proportion of amino acid substitutions at constrained (slowly evolving) sites after duplication was observed. An increase in the proportion of replacements at normally constrained amino acid sites could result from relaxation of purifying selective pressure. However, the proportion of amino acid replacements involving radical changes in amino acid properties after duplication does not appear to be significantly increased by relaxed selective pressure. The increased proportion of replacements at constrained sites was observed over a relatively large range of protein change (up to 25% amino acid replacements per site). These findings have implications for our understanding of the nature of evolution after duplication and may help to shed light on the evolution of novel protein functions through gene duplication. PMID- 12654936 TI - Messenger RNA surveillance and the evolutionary proliferation of introns. AB - The mechanisms responsible for the proliferation and subsequent stabilization of introns within the eukaryotic lineage have remained elusive. In the early stages of eukaryotic evolution, most introns may have been mildly deleterious at the time of insertion, but enough of them eventually acquired integral roles in transcript processing that few eukaryotic species can any longer survive without them. We suggest that the proliferation of spliceosomal introns was facilitated by the evolution of nonsense-mediated decay, an ancient and (in many cases) intron-dependent mechanism for eliminating aberrant mRNA molecules resulting from errors in transcription and splicing and from mutations at the DNA level. The spatial distribution of introns, as revealed by whole-genome analysis, is consistent with expectations for a model in which maximum protective coverage of a gene stochastically evolves over time. PMID- 12654937 TI - Recent horizontal transfer of mellifera subfamily mariner transposons into insect lineages representing four different orders shows that selection acts only during horizontal transfer. AB - We report the isolation and sequencing of genomic copies of mariner transposons involved in recent horizontal transfers into the genomes of the European earwig, Forficula auricularia; the European honey bee, Apis mellifera; the Mediterranean fruit fly, Ceratitis capitata; and a blister beetle, Epicauta funebris, insects from four different orders. These elements are in the mellifera subfamily and are the second documented example of full-length mariner elements involved in this kind of phenomenon. We applied maximum likelihood methods to the coding sequences and determined that the copies in each genome were evolving neutrally, whereas reconstructed ancestral coding sequences appeared to be under selection, which strengthens our previous hypothesis that the primary selective constraint on mariner sequence evolution is the act of horizontal transfer between genomes. PMID- 12654938 TI - The mammalian Crx genes are highly divergent representatives of the Otx5 gene family, a gnathostome orthology class of orthodenticle-related homeogenes involved in the differentiation of retinal photoreceptors and circadian entrainment. AB - The mammalian Crx genes are highly divergent orthodenticle (otd)-related homeogenes that play important roles in the differentiation of retinal photoreceptors and the circadian entrainment. However, their evolutionary origin and orthological relationships with other otd-related genes remain unclear. An orthology relationship of these genes with the highly conserved Otx5 genes identified in fish and amphibians, and also expressed in the eye and epiphysis, has been proposed previously but remains controversial. To test this hypothesis, we have identified Crx genes in a wide range of mammals, including three marsupials, and Otx5-related genes in a lizard, a turtle, and two archosaurs (crocodile and chick), as well as in the pufferfish. Phylogenetic analyses of the coding sequences show that the mammalian Crx genes are orthologous to the Otx5 related genes isolated in other gnathostomes. They also indicate that a duplication event has taken place in actinopterygians, after the splitting of the Cladistia, and that a relaxation of the structural constraints acting on the gene coding region has occurred early in the mammalian lineage. This process may be linked not only to the loss of ancestral Otx5/Crx functions during gastrulation or in the retinal pigmented epithelium, but also to the evolution of photic entrainment mechanisms in mammals. PMID- 12654939 TI - Clinical trials in the Journal of Pediatric Psychology: applying the CONSORT statement. AB - OBJECTIVE: To determine how well therapeutic randomized controlled trials (RCTs) in the Journal of Pediatric Psychology (JPP) met the CONSORT criteria as compared to pediatric trials in the Journal of Clinical and Consulting Psychology (JCCP), which served as a control. METHODS: Nine trials were found in JPP and 19 clinical trials were retrieved from JCCP. The modified Consort Checklist and Flow Chart was applied to each trial by two unblinded independent raters. RESULTS: The number of CONSORT items not reported between the two journals was strikingly similar. One half (11/22) of the CONSORT items were reported less than 25% of the time. Information provided on the flow of participants in the trials was similar across the two journals; however, JCCP more often reported on those assessed to be eligible overall, chi(2) = 5.241, p <.05. JPP reported on recruitment 33.3% of the time, while it was not reported at all in JCCP, chi(2) = 7.093, p <.05. CONCLUSIONS: Although many of the items in the CONSORT statement were not adhered to by the two psychological journals, most of the CONSORT items can clearly be applied to psychosocial trials. PMID- 12654940 TI - Commentary: the Journal of Pediatric Psychology should adopt the CONSORT statement as a way of improving the evidence base in pediatric psychology. PMID- 12654942 TI - Hemophilia Growth and Development Study: caregiver report of youth and family adjustment to HIV disease and immunologic compromise. AB - OBJECTIVE: To assess differences in caregiver report of youth and family psychosocial adjustment associated with HIV infection and greater immune compromise in youths with hemophilia. METHODS: Caregivers of 162 boys with hemophilia 8 to 20 years old completed three youth and family questionnaires (Personality Inventory for Children, Revised [PIC-R]; Questionnaire on Resources and Stress [QRS]; Family Environment Scale). RESULTS: Caregivers of HIV positive (HIV+) youths reported greater health concerns, social withdrawal (PIC-R), physical and adaptive limitations associated with illness (QRS) in their sons, and more pessimism about their sons' future and negative attitudes about parenting (QRS). Caregivers of HIV+ youths with greater immune compromise reported greater concerns about their sons' health and greater pessimism about their futures, as well as lower levels of family integration and more limited family opportunities. CONCLUSIONS: Results suggest caregivers perceive psychosocial problems in HIV+ youths with hemophilia and their families; some problems are specifically associated with greater immune compromise. PMID- 12654944 TI - Brief report: Cross-cultural replication of an anomalous psychometric pattern in children with type 1 diabetes. AB - OBJECTIVE: To replicate an anomalous psychometric profile previously documented in children with Type 1 diabetes living in the mainland United States with a cross-cultural sample selected from Puerto Rico. METHODS: Ninety-three Spanish speaking children (M age = 12.8 years) with Type 1 diabetes living in Puerto Rico were administered the Puerto Rican version of the Wechsler Intelligence Scale for Children-Revised (WISC-R). The factor structure of the Puerto Rican sample's WISC R was then compared to that of a United States sample (n = 95) in which an anomalous factor structure in children with diabetes was first documented. RESULTS: As in the United States sample, a four-factor IQ structure was obtained. Instead of the traditional three-factor structure of the WISC-R, the Perceptual Organization factor split into a Spatial Conceptual factor and an anomalous Visual Discrimination factor. CONCLUSIONS: Results support previous findings and suggest anomalies in the psychometric profiles of children with Type 1 diabetes. Cross-cultural replication of the anomalous IQ factor structure, and atypical visual discrimination, suggests that differences are illness-related, and consideration may therefore be warranted when administering some subtests of the Wechsler scales to children with Type 1 diabetes. PMID- 12654943 TI - Brief report: Parental perceptions of child vulnerability in children with chronic illness. AB - OBJECTIVE: To determine the extent to which parental perceptions of child vulnerability predict school and social adjustment in children with chronic illness. METHODS: Sixty-nine child-parent dyads were recruited from pediatric rheumatology and pulmonary clinics. Parents completed a self-report measure of parental perceptions of child vulnerability. Children completed measures of social adjustment. Parents also provided written permission to obtain school attendance records. Physicians provided a global assessment of children's disease severity. RESULTS: Increased parental perceptions of child vulnerability were related to increased social anxiety in children, even after controlling for child age and disease severity. Lower levels of parental education related to both increased perceptions of child vulnerability and increased school absences. CONCLUSIONS: Health providers should assess parental beliefs and parenting practices in assessing the adjustment of children with chronic illness. Moreover, interventions aimed at enhancing child adjustment to chronic illness might best target parents as well as children. PMID- 12654945 TI - Stress and adaptation in mothers of children with cerebral palsy. AB - OBJECTIVE: To assess the importance of disability severity and child functional status as predictors of maternal depressive symptoms and the moderating effects of maternal appraisal, social support, and family income on the relationship between disability factors (severity and functional status) and maternal depressive symptoms. METHOD: Mothers of 270 children with cerebral palsy completed surveys on their appraisal of the child's disability, social support, and family demographics. Physicians assessed the severity of the disability and the child's functional status. RESULTS: Thirty percent of the mothers had depressive symptoms above the cutoff on a depression screening instrument. Disability severity and child's functional status did not predict maternal depression. Perceived social support moderated the relationship between the child's functional status and maternal depressive symptoms. CONCLUSIONS: Mothers of children with cerebral palsy may be at risk for depression. Interventions that take into account the moderating effects of social support may increase maternal adaptation. PMID- 12654946 TI - Blood glucose estimations in adolescents with type 1 diabetes: predictors of accuracy and error. AB - OBJECTIVE: To examine predictors of blood glucose (BG) estimation accuracy and errors in adolescents with Type 1 diabetes. METHOD: Seventy-eight adolescents (ages 11-19) rated their physical symptoms and estimated their BG prior to conducting daily BG tests. BG estimation data were subject to an error grid analysis. Hierarchical regression was used to identify predictors of BG estimation accuracy and errors. RESULTS: The average participant made accurate BG estimations 37% of the time and clinically relevant BG estimation errors 24% of the time. Girls and older adolescents had higher BG estimation accuracy rates and lower BG estimation error rates than boys and younger adolescents. Higher BG variability was also associated with increased BG estimation errors. CONCLUSIONS: Although boys, younger adolescents, and those with higher BG variability showed higher BG estimation error rates, most participants showed low rates of BG estimation accuracy and high rates of BG estimation error. Health providers may underestimate the frequency of inappropriate self-treatment as a result of patient difficulty in estimating BG accurately. PMID- 12654947 TI - Interactions between children with juvenile rheumatoid arthritis and their mothers. AB - OBJECTIVE: To determine the degree to which mothers of children with juvenile rheumatoid arthritis (JRA) show an overprotective or highly controlling interaction style. METHOD: We videotaped 84 mother-child pairs (42 JRA and 42 healthy, ages 6 to 13) while working on a collaborative problem-solving task. Based on physical therapy evaluations, children in the JRA group were assigned to "more severe" (n = 19) and "milder" (n = 22) arthritis subgroups. RESULTS: Results showed numerous differences between mothers of children with more severe arthritis and the other mothers (no differences between the milder arthritis and healthy comparison groups were found). Compared to mothers in the other two groups, mothers of children with more severe arthritis were more directive of their children's behavior during the task, showing higher rates of structure and rule setting, general clues, and prompting the child for an answer. DISCUSSION: Sequential analyses showed that mothers in the more severe group appeared to treat the task in a more evaluative manner, being more likely than other mothers to respond to correct answers with positive feedback and to incorrect answers with structure and rule setting. Mothers in the other groups were more likely to respond to both correct and incorrect answers with specific clues. CONCLUSIONS: We discuss how these differences in interactional style might impact the social development of children with JRA. PMID- 12654948 TI - Progressive supranuclear palsy: Setting the scene for therapeutic trials. PMID- 12654949 TI - An old acquaintance: opioids in neuropathic pain. PMID- 12654950 TI - Hypnic headache: clinical features, pathophysiology, and treatment. AB - Hypnic headache has been described in several case reports since 1981 and is regarded as an idiopathic headache disorder. In this review of 71 cases in the literature, the clinical features, neurophysiologic including polysomnographic findings, and treatment procedures are analyzed and the pathophysiology of this condition, which remains however speculative, is discussed. There is some evidence that hypnic headache is related to REM sleep. The analysis shows that hypnic headache most probably is an entity among the idiopathic headache disorders unassociated with structural lesions and does not belong to the trigeminal-autonomic cephalalgias. Lithium shows the best efficacy; indomethacin, flunarizine, and caffeine may also be useful. PMID- 12654951 TI - Quantitative sensory testing: report of the Therapeutics and Technology Assessment Subcommittee of the American Academy of Neurology. AB - OBJECTIVE: This assessment evaluates the clinical utility, efficacy, and safety of quantitative sensory testing (QST). METHODS: By searching MEDLINE, Current Contents, and their personal files, the authors identified 350 articles. Selected articles utilized computer operated threshold systems, manually operated threshold systems, and electrical threshold devices. The authors evaluated the use of normal values and the degree of reproducibility between the same and different systems. Articles were rated using a standard classification of evidence scheme. RESULTS: Because of differences between systems, normal values from one system cannot be transposed to others. Reproducibility of results was also an important concern, and there is no consensus on how it should be defined. The authors identified no adequately powered class I studies demonstrating the effectiveness of QST in evaluating any particular disorder. A number of class II and III studies demonstrated that QST is probably or possibly useful in identifying small or large fiber sensory abnormalities in patients with diabetic neuropathy, small fiber neuropathies, uremic neuropathies, and demyelinating neuropathy. CONCLUSIONS: QST is a potentially useful tool for measuring sensory impairment for clinical and research studies. However, QST results should not be the sole criteria used to diagnose pathology. Because malingering and other nonorganic factors can influence the test results, QST is not currently useful for the purpose of resolving medicolegal matters. Well-designed studies comparing different QST devices and methodologies are needed and should include patients with abnormalities detected solely by QST. PMID- 12654952 TI - Clinical features and natural history of progressive supranuclear palsy: a clinical cohort study. AB - OBJECTIVE: To describe clinical features and identify prognostic predictors in progressive supranuclear palsy (PSP). METHODS: Record-based diagnosis according to National Institute of Neurological Disorders and Stroke-Society for Progressive Supranuclear Palsy criteria was performed in 187 cases of PSP. Clinical information was abstracted from patient records. Sixty-two patients (33%) were examined by the investigators. Forty-nine of 62 patients (79%) underwent standardized clinical assessment. Predictors of survival were examined after a mean of 6.4 years. RESULTS: The most common symptoms at disease onset related to mobility (69%). Of patients undergoing standardized clinical assessment, diplopia occurred in 61%, photophobia in 43%, and eyelid apraxia in 43%. Seventy-five cases (40%) died during follow-up. Older age at onset and classification as probable PSP were associated with poorer survival. Onset of falls (hazard ratio 3.28, 95% CI 1.17 to 9.13), speech problems (hazard ratio 4.74, 95% CI 1.10 to 20.4), or diplopia (hazard ratio 4.23, 95% CI 1.23 to 14.6) within 1 year and swallowing problems within 2 years (hazard ratio 3.91, 95% CI 1.39 to 11.0) were associated with a worse prognosis. CONCLUSIONS: Mobility problems are the commonest early feature in PSP and visual symptoms are often functionally disabling. Early falls, speech and swallowing problems, diplopia, and early insertion of a percutaneous gastrostomy predict reduced survival. PMID- 12654953 TI - Progression of gait, speech and swallowing deficits in progressive supranuclear palsy. AB - OBJECTIVE: To identify outcome measures for clinical trials in progressive supranuclear palsy (PSP), the authors determined the time to key motor impairments in a well-defined patient cohort. METHODS: The records of consecutive patients with probable PSP, defined by National Institute of Neurological Disorders and Stroke-Society for Progressive Supranuclear Palsy workshop criteria on first visit to a tertiary care center, were selected for study if the patients were seen regularly at 3- to 6-month intervals, had at least 24 months of care at the Rush movement disorder center, and were originally seen in the movement disorder center before any of the following key motor impairments developed: unintelligible speech, no independent walking, inability to stand unassisted, wheelchair-bound, or recommendation for feeding tube placement. Using standardized criteria from the Unified Parkinson's Disease Rating Scale and Hoehn and Yahr stages taken at each clinic visit, the authors recorded the time that each patient reached the key motor impairments and death. Median months from first symptom onset and from first consultation at the movement disorder center to each motor impairment were derived from Kaplan-Meier curves. RESULTS: Fifty subjects, mean age at study entry 64.2 years, were followed to death (n = 21, mean duration of surveillance 53.6 months) or in ongoing fashion (n = 29, mean duration of surveillance 46.2 months). Eighty-eight percent of the sample met at least one milestone. The median time from disease onset to the first key motor impairment was 48 months, 24 months after first consultation. The three gait items occurred temporally close, and the authors considered them thereafter as a single milestone, occurring at a median disease duration of 57 months, 34 months after first consultation. Unintelligible speech occurred at a median disease duration of 71 months, median 44 months after first consultation. As a composite end point, speech/gait accounted for 98% of the sample's first key motor impairment. Need for nasogastric tube was rarely the first milestone (8%). CONCLUSIONS: Gait impairment and unintelligible speech are milestones that occur rapidly in PSP and can be monitored with standardized rating scales. The authors suggest that clinical trials use these indices to assess how interventions alter anticipated progression of clinical deterioration in PSP. PMID- 12654954 TI - Diffusion-weighted imaging discriminates progressive supranuclear palsy from PD, but not from the parkinson variant of multiple system atrophy. AB - BACKGROUND AND OBJECTIVE: The parkinson variant of multiple system atrophy (MSA P) and progressive supranuclear palsy (PSP) present with atypical parkinsonism, which may be misdiagnosed as PD, particularly in early disease stages. It was previously shown that diffusion-weighted MRI (DWI) is a sensitive tool to discriminate MSA-P from PD based on increased apparent diffusion coefficients (ADCs) in the putamen. In this study DWI was evaluated in 10 patients with PSP compared with 13 patients with PD and 12 with MSA-P. METHODS: Disease was diagnosed according to established diagnostic criteria and groups were matched for age, disease duration, and Hoehn and Yahr "off" stage. Regional ADCs (rADCs) were determined in different brain regions including basal ganglia, gray matter, white matter, substantia nigra, and pons. RESULTS: In patients with PSP compared with those with PD, rADCs were significantly increased in putamen, globus pallidus, and caudate nucleus. Stepwise logistic regression analysis followed by receiver operating characteristics analysis identified an optimal cut-off value for putaminal rADC, discriminating PSP and PD with a sensitivity of 90% and a positive predictive value of 100%. DWI failed to discriminate PSP and MSA-P. CONCLUSIONS: These results show that DWI detects basal ganglia abnormalities in PSP patients within few years of disease onset, discriminating patients with PSP from those with PD, but not from those with MSA-P. PMID- 12654955 TI - Controlled-release oxycodone for pain in diabetic neuropathy: a randomized controlled trial. AB - BACKGROUND AND OBJECTIVE: Opioid treatment has played a limited role in the management of diabetic neuropathy, in part because of concerns about the responsiveness of neuropathic pain to opioid treatment. This controlled study evaluated the efficacy and safety of controlled-release (CR) oxycodone in subjects with moderate to severe pain due to diabetic neuropathy. METHODS: This multicenter, randomized, double-blind, placebo-controlled, parallel-group study included 159 subjects with moderate to severe pain due to diabetic neuropathy. Treatment began with either one 10-mg tablet of CR oxycodone (n = 82) or identical placebo (n = 77) every 12 hours. Doses could be increased every 3 days to a maximum of 6 tablets (60 mg CR oxycodone) every 12 hours. Treatment lasted up to 6 weeks. The primary efficacy variable was overall average daily pain intensity during study days 28 to 42. RESULTS: At an average (SD) dose of 37 (21) mg per day (range 10 to 99 mg/d), CR oxycodone provided more analgesia than placebo (p= 0.002) in the intent-to-treat cohort. From days 28 to 42, overall average daily pain intensity (least squares mean +/-SE), rated in subject diaries on a numeric scale of 0 (no pain) to 10 (pain as bad as you can imagine), was 4.1 +/- 0.3 in subjects given CR oxycodone and 5.3 +/- 0.3 in placebo-treated subjects. Overall, 80 (96%) of 82 subjects given CR oxycodone and 52 (68%) of 77 subjects who received placebo reported adverse events. The most common adverse events in the CR oxycodone group were opioid related. CONCLUSIONS: In this 6-week trial, CR oxycodone was effective for the treatment of moderate to severe pain due to diabetic neuropathy. Adverse events were typical of opioid-related side effects. PMID- 12654956 TI - Premonitory symptoms in migraine: an electronic diary study. AB - BACKGROUND: Migraine is frequently associated with nonheadache symptoms before, during, and after the headache. Premonitory symptoms occurring before the attack have not been rigorously studied. Should these symptoms accurately predict headache, there are considerable implications for the pathophysiology and management of migraine. METHODS: Electronic diaries were used in a 3-month multicenter study to record nonheadache symptoms before, during, and after migraine. The authors recruited subjects who reported nonheadache symptoms in at least two of three attacks that they believed predicted headache. Symptoms were entered in the diaries by patient initiation and through prompted entries at random times daily. Entries could not be altered retrospectively. Data recorded included nonheadache symptoms occurring during all three phases of the migraine, prediction of the attack from premonitory symptoms, general state of health, and action taken to prevent the headache. RESULTS: One hundred twenty patients were recruited: 97 provided usable data. Patients correctly predicted migraine headaches from 72% of diary entries with premonitory symptoms. A range of cognitive and physical symptoms was reported at a similar rate through all three phases of the migraine. The most common premonitory symptoms were feeling tired and weary (72% of attacks with warning features), having difficulty concentrating (51%), and a stiff neck (50%). Subjects who functioned poorly in the premonitory phase were the most likely to correctly predict headache. CONCLUSIONS: Using an electronic diary system, the authors show that migraineurs who report premonitory symptoms can accurately predict the full-blown headache. PMID- 12654957 TI - CSF hypovolemia vs intracranial hypotension in "spontaneous intracranial hypotension syndrome". AB - OBJECTIVES: To investigate the role of CSF hypovolemia in spontaneous intracranial hypotension (SIH) syndrome because so-called SIH syndrome sometimes lacks intracranial hypotension. METHODS: Ten women (aged from 28 to 49 years) with characteristic orthostatic headache without a previous history of dural tear were investigated. In addition to gadolinium (Gd)-enhanced brain MRI, spinal MRI with and without Gd enhancement was performed. RESULTS: Gd-enhanced brain MRI demonstrated diffuse pachymeningeal enhancement in all patients. Sagittal T2 weighted spinal MRI revealed a variable amount of CSF in the extradural space in all patients. Sagittal T2-weighted MRI or axial Gd-enhanced T1-weighted MRI showed dilated epidural veins located in the high cervical portion in each patient. The intensity of dilatation of the epidural veins correlated significantly with the amount of CSF in the epidural space. This suggested that the Monro-Kellie doctrine was applicable in this circumstance. CONCLUSIONS: Since some patients with SIH syndrome have normal CSF pressure and since a downward displacement of the brain due to a reduction of the buoyant action of CSF may induce symptoms, CSF hypovolemia, not intracranial hypotension, may be the cause. Based on the Monro-Kellie doctrine, detecting leaked CSF and venous engorgement (epidural vein dilatation and pachymeningeal enhancement) is an important clue to diagnose so-called SIH syndrome. Dilatation of epidural veins suggests CSF hypovolemia in appropriate conditions. PMID- 12654958 TI - Protein S-100b serum levels in trauma-induced brain death. AB - OBJECTIVE: To analyze the time course of serum protein S-100b in patients with traumatic brain injury deteriorating to brain death and to investigate the predictive value of initial S-100b levels in relation to clinical and radiologic measures of injury severity with regard to brain death. METHODS: Forty-seven patients who sustained severe head injury were studied. Blood samples for measurement of S-100b were drawn on admission in the intensive care unit and every 24 hours thereafter for a maximum of 6 consecutive days or until brain death occurred. Variables related to outcome were recorded, including age, sex, Glasgow Coma Scale (GCS), and brain CT findings on admission. Outcome was defined as deterioration to brain death or not. RESULTS: Of the 47 patients studied, 17 deteriorated to brain death and 30 did not. On admission, patients who became brain dead had higher median serum S-100b levels compared with those who did not (2.32 microg/L vs 1.04 micro g/L, p = 0.0028). Logistic regression analysis showed that initial S-100b was an independent predictor of brain death (p = 0.041), in the presence of advanced age (p = 0.043) and low GCS score (p = 0.013). The odds ratio of 2.09 (95% CI, 1.03 to 4.25) indicates a more than doubling of the probability of deteriorating to brain death per 1- micro g/L increase in S-100b on admission. At clinical brain death, median S-100b was higher in patients with brain death compared with the peak S-100b value obtained over a 6-day period in those who did not become brain dead (6.58 microg/L vs 1.49 microg/L, p < 0.0001). CONCLUSIONS: Prediction of brain death after severe head injury can be improved by combining clinical and S-100b data; thus, serum S-100b determination deserves to be included in the neuromonitoring of patients with severe traumatic brain injury. PMID- 12654959 TI - Early and late cognitive changes following temporal lobe surgery for epilepsy. AB - OBJECTIVE: To evaluate early and late postoperative cognitive changes in adult patients who had a standardized temporal lobe resection for control of intractable complex partial seizures and to relate quality of life measures at the long-term follow-up to seizure control and a verbal memory skill. METHODS: Patients in this longitudinal study had intractable epilepsy and had cognitive tests at the initial surgical evaluation, 1 year later, and at a long-term follow up (at least >9 years later). Mean follow-up was 12.8 years. Participants were 44 patients who had an en bloc left (LTL) or right (RTL) temporal lobe resection and 8 nonsurgical patients with epilepsy. RESULTS: Patients with LTL surgery showed selective early decreases in verbal memory. At the long-term follow-up, further decreases in verbal memory and visual memory scores were seen for all patient groups. In general, the nonmemory scores remained stable over time. LTL surgery and initial high scores were predictors of verbal memory decreases seen at the early follow-up. Late memory declines were predicted by higher 1-year scores. LTL surgery was an additional predictor of late decline on a verbal memory task sensitive to the integrity of the left hippocampus. Higher quality of life scores of patients at the long-term follow-up were associated with both better seizure control and a higher everyday verbal memory skill: prose recall. CONCLUSIONS: Surgery-induced verbal memory deficits following LTL surgery continue 13 years after surgery. Late, possibly age-related, declines in multiple memory scores are seen in patients with temporal lobe surgery and nonsurgical patients with epilepsy. Patients with LTL surgery may be at risk for a more rapid decline in selective verbal memory skills. Evaluations of treatments for intractable epilepsy that compromise memory functions should consider the further quality of life impact of late age-related memory declines. PMID- 12654960 TI - Alpha-methyl-L-tryptophan PET detects epileptogenic cortex in children with intractable epilepsy. AB - BACKGROUND: In children with tuberous sclerosis, the PET tracer alpha[11C]methyl L-tryptophan (AMT) has been shown to be selectively taken up by epileptogenic tubers, thus allowing differentiation from nonepileptogenic tubers in the interictal state. OBJECTIVE: To determine whether cortical areas showing increased AMT uptake in children without tuberous sclerosis complex with intractable neocortical epilepsy indicate the epileptogenic zone, and to assess the relative contributions of AMT and 2-deoxy-2[18F]fluoro-D-glucose (FDG) PET abnormalities to the localization of epileptogenic cortical regions. METHODS: Areas of increased AMT and decreased FDG uptake were marked objectively as regions with abnormal asymmetry using an in-house written software in 27 children who underwent comprehensive evaluation for resective epilepsy surgery. The marked PET abnormalities were compared to the locations of scalp and subdural EEG epileptiform abnormalities, as well as histology and surgical outcome. RESULTS: Focal cortical increases of AMT uptake were found in 15 patients. The lobar sensitivity (39.0%) of AMT PET for seizure onset was lower, but its specificity (100%) was higher (p < 0.0001) than that of hypometabolism on FDG PET (sensitivity 73.2%, specificity 62.7%). AMT PET abnormalities were smaller than corresponding FDG PET hypometabolic regions (p = 0.002), and increased AMT uptake occurred in two patients with nonlocalizing FDG PET. Histologically verified cortical developmental malformations were associated with increased AMT uptake (p = 0.044). Subdural electrodes adjacent to the area of increased AMT uptake were most often involved in seizure onset. CONCLUSIONS: Focal increase of cortical AMT uptake in children is less sensitive but more specific for the lobe of seizure onset than corresponding FDG PET hypometabolism, and it is often associated with epileptogenic cortical developmental malformations. AMT PET can assist placement of subdural electrodes even when MRI and FDG PET fail to provide adequate localizing information. Cortical areas adjacent to increased AMT uptake should be carefully addressed by intracranial EEG because these regions often show a high degree of epileptogenicity. PMID- 12654961 TI - Intrasubject reproducibility of presurgical language lateralization and mapping using fMRI. AB - BACKGROUND: fMRI is becoming a standard tool for the presurgical lateralization and mapping of brain areas involved in language processing. However, its within subject reproducibility has yet to be fully explored. OBJECTIVE: To evaluate within-test and test-retest reliability of language fMRI in consecutive patients undergoing evaluation for epilepsy surgery. METHODS: Thirty-four unselected patients were investigated once (within-test reliability) and 12 patients twice (test-retest reliability). The imaging series consisted of an alternating 25 second synonym judgment condition with a 25-second letter-matching condition repeated 15 times. Reproducibility of activation maps of the first and second half of session 1 or activation maps of sessions 1 and 2 was evaluated by comparing one global and three regional lateralization indexes (Broca's area, remaining prefrontal cortex, temporoparietal area) and on a voxel-by-voxel basis (intraclass correlation coefficient, percentage overlap, correlation of t values). RESULTS: Global and regional language lateralization was achieved with high reliability within and across sessions. Reproducibility was evenly distributed across both hemispheres but not within each hemisphere. Frontal activations were more reliable than temporoparietal ones. Depending on the statistical threshold chosen, the voxel-by-voxel analysis revealed a mean overlap of activations derived from the first and second investigation of up to 48.9%. CONCLUSION: Language fMRI proved sufficiently reliable for the determination of global and regional lateralization of language representation in individual unselected patients with epilepsy. PMID- 12654962 TI - Occurrence of seizure clusters and status epilepticus during inpatient video-EEG monitoring. AB - OBJECTIVE: To investigate the occurrence of status epilepticus and seizure clusters, and the duration until first seizure at epilepsy monitoring units in the United States. METHODS: The authors examined the inpatient video-EEG monitoring reports of 514 consecutive patients admitted to five comprehensive epilepsy centers during the year 2000. Time to first seizure, seizure clustering, and seizure duration were ascertained from reports and entered into a database. RESULTS: In 169 admissions with complex partial seizures (CPSs) or secondarily generalized tonic-clonic (2GTC) seizures, there were 5 (3.0%) patients with status epilepticus, 30 (17.8%) with 4-hour seizure clusters, and 82 (48.5%) with 24-hour seizure clusters. There were no statistically significant differences between centers, except that seizure clusters were observed to be less common at the one center with a formal drug withdrawal protocol. The average time to CPS or 2GTC seizure was 2.1 days; the average number of days to nonepileptic event was 1.2 days (p = 0.001). CONCLUSIONS: Although status epilepticus is uncommon at epilepsy monitoring units, clusters of seizures are common. Intensive monitoring with drug withdrawal must be performed in a highly supervised, hospitalized setting. Inpatient video-EEG monitoring is efficient, with recording of the first epileptic or nonepileptic events in 2 days or less. PMID- 12654963 TI - The accuracy of the diagnosis of paroxysmal events in children. AB - OBJECTIVE: To assess the accuracy of the diagnosis of epileptic seizures in children. METHODS: The Dutch Study of Epilepsy in Childhood is a prospective hospital-based study of 881 children referred because of possible seizures. The diagnosis was based on predefined descriptive criteria, as applied by a panel of three pediatric neurologists. Children with a definite other diagnosis were excluded. All children with unclear events were followed up for 1 year and children with seizures were followed up for 2 years to assess the accuracy of the diagnosis. RESULTS: In 170 of 224 children seen after a single event, the incident was classified initially as epileptic, in 54 as unclear. In none of the 170 children did the diagnosis prove to be wrong. In four of the 54 children, recurrent episodes enabled a definite diagnosis of epilepsy. In 412 of the 536 children seen with multiple events, an initial diagnosis of epilepsy was made. After follow-up, this initial diagnosis was probably incorrect in 19. In contrast, seven of 124 children with multiple unclear episodes at intake later received the diagnosis epilepsy. CONCLUSIONS: A false-positive diagnosis of epilepsy was made in 4.6%, whereas a definite diagnosis of epilepsy or seizure was delayed in 5.6% of children with multiple unclear events and in 7.4% of children with one unclear event. PMID- 12654964 TI - Classical infantile spinal muscular atrophy with SMN deficiency causes sensory neuronopathy. AB - OBJECTIVE: Classic infantile spinal muscular atrophy (SMA) is believed to be a purely motor disorder, affecting neurons of the spinal anterior horn and nuclei of the lower cranial nerves. Other organ malformations or peripheral nerve involvement have been regarded as exclusion criteria for infantile SMA. Whether SMN protein deficiency can also lead to loss of sensory neurons has not been systematically addressed. METHODS: The authors evaluated the sural nerve biopsies of 19 patients with infantile SMA of varying severity. The diagnosis of SMA was confirmed by the presence of a homozygous deletion of the SMN1 gene in all patients. RESULTS: In seven unrelated infants with SMA type I, axonal degeneration of the sural nerve was noted. Five patients showed abnormal sensory conduction, thus prompting sural nerve biopsy. Sural nerves showed different degrees of axonal loss: fiber density ranged from 3.482 to 22.076/mm2 and was markedly reduced in four patients. There was no evidence of primary demyelination: the ratio of total myelinated fiber thickness to axon diameter (g ratio) was normal in the patients examined. In seven patients with SMA II and five patients with SMA III, no sural nerve alterations were seen, and conduction velocity was normal. In addition to SMN1 gene deletions, homozygous NAIP gene deletions were detected in six out of seven infants with peripheral neuropathy, whereas there was no evidence of a large deletion including the multicopy markers C212 and Ag1-CA in two out of three families tested. CONCLUSIONS: In this series of patients with SMA I through III who underwent sural nerve biopsy, there was significant sensory nerve pathology in severely affected patients with SMA type I, whereas there were no sensory nerve alterations clinically or morphologically in patients with milder SMA type II or III. PMID- 12654966 TI - Activation of nuclear factor-kappaB in inflammatory myopathies and Duchenne muscular dystrophy. AB - OBJECTIVE: To investigate the immunolocalization and activation of nuclear factor kappaB (NF-kappaB) in polymyositis, dermatomyositis, and Duchenne muscular dystrophy (DMD). BACKGROUND: NF-kappaB is a major transcription factor modulating the cellular immune, inflammatory, and proliferative responses. In skeletal muscle it was demonstrated to play a role in the expression of inducible genes in response to oxidative stress and ischemia/reperfusion injury, and also in myonuclear apoptosis and muscle catabolism. Some data suggest that NF-kappaB may play a role in the pathogenesis of inclusion body myositis. METHODS: Muscle samples from five patients each with polymyositis, dermatomyositis, and DMD and 10 normal controls were studied by immunocytochemistry and Western blot of nuclear extracts for the activated form of NF-kappaB. NF-kappaB DNA binding activity was studied by electrophoretic mobility shift assay (EMSA). RESULTS: Immunoreactivity for NF-kappaB was found in the cytoplasm of all regenerating fibers and in 20 to 40% of necrotic fibers. Western blot analysis of nuclear extracts showed a single band corresponding to 65 kd in all patients. EMSA analysis confirmed activation of NF-kappaB pathway in inflammatory myopathies and, to a lesser extent, also in DMD. CONCLUSIONS: These data indicate that nuclear factor-kappaB pathway is activated in polymyositis, dermatomyositis, and Duchenne muscular dystrophy. It may play a role in modulating the immune response and in regulating myogenesis and muscle repair. PMID- 12654965 TI - FKRP gene mutations cause congenital muscular dystrophy, mental retardation, and cerebellar cysts. AB - BACKGROUND: Congenital muscular dystrophies (CMD) are autosomal recessive disorders that present within the first 6 months of life with hypotonia and a dystrophic muscle biopsy. CNS involvement is present in some forms. The fukutin related protein gene (FKRP) is mutated in a severe form of CMD (MDC1C) and a milder limb girdle dystrophy (LGMD2I). Both forms have secondary deficiencies of laminin alpha2 and alpha-dystroglycan immunostaining. Structural brain involvement has not been observed in patients with FKRP gene mutations. METHODS: The authors studied two unrelated patients who had a pattern of muscle involvement identical to MDC1C, mental retardation, and cerebellar cysts on cranial MRI. The FKRP gene was analyzed along with the skeletal muscle expression of laminin alpha2 and alpha-dystroglycan. RESULTS: The muscle biopsy of both patients showed severe dystrophic findings, a reduction in laminin alpha2, and profound depletion of alpha-dystroglycan. Both patients had homozygous FKRP gene mutations not previously reported (C663A [Ser221Arg] and C981A [Pro315Thr]). CONCLUSIONS: Mutations within the FKRP gene can result in CMD associated with mental retardation and cerebellar cysts. This adds structural brain defects to the already wide spectrum of abnormalities caused by FKRP mutations. The severe depletion of alpha-dystroglycan expression suggests that FKRP is involved in the processing of alpha-dystroglycan. PMID- 12654967 TI - Predictors of nursing home placement in Huntington disease. AB - OBJECTIVE: To determine whether motor, behavioral, or psychiatric symptoms in Huntington disease (HD) predict skilled nursing facility (SNF) placement. METHODS: Subjects were participants in the Huntington Study Group's Unified Huntington Disease Rating Scale Database (Rochester, NY) between January 1994 and September 1999. Specific motor, psychiatric, and behavioral variables in subjects residing at home and in SNF were analyzed using chi2 and Student's t-tests. For a subset of subjects for whom longitudinal data existed, a Cox proportional hazards model controlling for age, sex, and disease duration was used. RESULTS: Among 4,809 subjects enrolled, 3,070 had clinically definite HD. Of these, 228 (7.4%) resided in SNF. The SNF residents' average age was 52 years, average disease duration was 8.6 years, and they were predominantly women (63%). The SNF residents had worse motor function (chorea, bradykinesia, gait abnormality, and imbalance, p < 0.0001); were more likely to have obsessions, compulsions, delusions, and auditory hallucinations; and had more aggressive, disruptive (p < 0.0001), and irritable behaviors (p = 0.0012). For 1,559 subjects, longitudinal data existed (average length of follow-up, 1.9 years), and 87 (5%) moved from home to SNF. In the Cox model, bradykinesia (HR 1.965, 95% CI 1.083 to 3.564), impaired gait (HR 3.004, 95% CI 1.353 to 6.668), and impaired tandem walking (HR 2.546, 95% CI 1.460 to 4.439) were predictive of SNF placement. CONCLUSIONS: Institutionalized patients with HD are more motorically, psychiatrically, and behaviorally impaired than their counterparts living at home. However, motor variables alone predicted institutionalization. Treatment strategies that delay the progression of motor dysfunction in HD may postpone the need for institutionalization. PMID- 12654968 TI - Strong association between N-acetyltransferase 2 genotype and PD in Hong Kong Chinese. AB - BACKGROUND: The slow acetylator genotype for N-acetyltransferase 2 (NAT2 genotype) may be associated with PD in white subjects and the genotype is common in both white and Chinese populations. Whether there is a relationship between NAT2 genotype and PD in Chinese subjects is not known. OBJECTIVE: To investigate the association between the slow acetylator genotype for N-acetyltransferase 2 and PD in a Chinese population. METHODS: The authors obtained DNA samples and documented sex, age, and smoking history in 99 Chinese patients with PD and in 126 control subjects from two major Hong Kong hospitals. PCR-restriction fragment length polymorphism was used to identify M1, M2, and M3 mutant polymorphisms of the slow acetylator genotype for N-acetyltransferase 2. Logistic regression analyses were carried out to investigate the relationships between the different variables and PD. RESULTS: The frequency of the slow acetylator genotype for N acetyltransferase 2 in the PD group was significantly higher than that of the control group (68.7% vs 28.6%) with an OR of 5.53 (95% CI 3.08 to 9.92) after adjusting for age, sex, and smoking history. In a subgroup analysis smoking had no modifying effect on the association between genotype and PD. CONCLUSIONS: There is a significant association between PD and the slow acetylator genotype for N-acetyltransferase 2 in Hong Kong Chinese. The OR found is among the highest reported so far in all susceptibility gene studies for PD in both Chinese and white subjects and provides evidence for a possible functional relationship between NAT2 slow acetylator genotype and PD in both racial groups. PMID- 12654969 TI - Staging disease severity in pathologically confirmed cases of frontotemporal dementia. AB - OBJECTIVE: To devise a staging scheme for addressing the severity of atrophy in patients with pathologically proven frontotemporal dementia (FTD) and determine any relationship with clinical indices. METHODS: Twenty-four cases with clinical and pathologic features of FTD were selected using standard inclusion and exclusion criteria from 125 dementia cases collected in Sydney, Australia, over an 8.5-year period. Patterns of gross atrophy were determined in two coronal brain slices. Reproducibility of a four-stage severity scheme was tested. Nonparametric statistics were used to determine relationships between the stage of atrophy and clinical indices (age at death, duration from diagnosis, and clinical severity at death). RESULTS: The FTD cases studied could be reliably grouped (kappa = 0.97) into four progressively severe stages of global atrophy. Initial mild atrophy occurred in the orbital and superior medial frontal cortices and hippocampus (stage 1), progressed to involve the other anterior frontal regions, temporal cortices, and basal ganglia (stage 2), then involved all remaining tissue in these coronal slices (stage 3), until very marked atrophy was observed in all areas (stage 4). These stages correlated with disease duration and clinical dementia severity, lending validity to the progressive nature of the staging scheme. CONCLUSIONS: The authors have identified a reproducible staging system for the severity of gross atrophy in cases of FTD. This staging scheme provides the required framework to compare different research indices and determine correlates relating to time and disease progression in FTD-information necessary to determine core disease processes and etiologic factors. PMID- 12654971 TI - James Glaisher's 1862 account of balloon sickness: altitude, decompression injury, and hypoxemia. AB - In 1862, James Glaisher and Henry Coxwell ascended to 29,000 feet in an open hot air balloon. During the ascent, Glaisher described marked neurologic compromises: appendicular and later truncal paralysis, blindness, initially preserved cognition, and subsequent loss of consciousness. The author examines Glaisher's account of balloon sickness by comparing it with other balloonists' observations and discussing it in the context of altitude sickness, decompression injury, and hypoxemia. PMID- 12654970 TI - Primary cervical dystonia and scoliosis: a multicenter case-control study. AB - OBJECTIVE: To study the distribution of prior scoliosis among patients with primary adult-onset cervical dystonia (CD) and matched control subjects. METHODS: Case and control subjects were selected among consecutive outpatients attending four Italian centers. Control outpatients were matched for age (+/-5 years), sex, and referral center. Information on prior scoliosis, other spine diseases, and family history of dystonia was obtained by a standardized questionnaire and supported by medical records. Conditional logistic regression models were used to adjust simultaneously for age, disease duration, and education level and to determine the independent association of exposure variables with the outcome. RESULTS: Prior scoliosis developing in middle or late childhood or at around the puberty occurred more frequently among 72 case patients than among 144 neurologic control subjects. No subject reported conditions considered to be responsible for secondary scoliosis. The association of scoliosis and CD was not confounded by age, duration of disease, education level, other spine diseases, or family history of dystonia (adjusted odds ratio [OR] 6.8; 95% CI 1.5 to 29.5; p = 0.011). The OR of family history of dystonia (18.7; 95% CI 2.4 to 147.5; p = 0.005) fell to 11.7 (95% CI 1.3 to 103; p = 0.03) after controlling for scoliosis. CONCLUSIONS: Prior scoliosis may increase the risk of developing CD. The observed decrease in the magnitude of the association between family history of dystonia and CD after controlling for scoliosis suggests a link between the two conditions. PMID- 12654972 TI - MRI in diabetic muscle infarction. PMID- 12654973 TI - Hemorrhagic stroke associated with the Iowa amyloid precursor protein mutation. AB - The authors searched for mutations in the beta-amyloid precursor protein in a Spanish family with a hereditary syndrome of hemorrhagic stroke, dementia, leukoencephalopathy, and occipital calcifications. DNA from two affected members demonstrated the Iowa amyloid precursor protein mutation previously identified as a cause of severe amyloid angiopathy without hemorrhagic stroke. These data point to other genetic or environmental factors that may determine the occurrence of symptomatic hemorrhage in amyloid angiopathy. PMID- 12654974 TI - APOE genotype is a risk factor for neuropathy severity in diabetic patients. AB - This cross-sectional study tested the hypothesis that APOE genotype is a risk factor for diabetic neuropathy severity. A model with age, duration of diabetes, and APOE genotype was found to predict (p = 0.0083) severity on the Neuropathy Impairment Score in the Lower Limbs (NISLL). Considering genotype alone, patients with APOE 3/4 and 4/4 genotypes had 3 more NISLL points than patients with other genotypes. This impact on severity is equivalent to having 15 extra years of age or diabetes duration. PMID- 12654975 TI - Myasthenia gravis: a higher than expected incidence in the elderly. AB - This 10-year (1991 to 2000) prospective study of MG in the county of Osona (Barcelona, Spain) reveals an annual incidence rate of 21.27 cases per million inhabitants (95% CI 13.89 to 31.16). Incidence increased from 5.03 x 10(6) in the age group of 0 to 14 years to 14.68 x 10(6) in the age group of 15 to 64 years and to 63.38 x 10(6) in the older population. These results, the highest reported to date, may be explained by the population aging. PMID- 12654976 TI - Elevated polyunsaturated fatty acids in blood serum obtained from children on the ketogenic diet. AB - The authors analyzed blood metabolites in nine children with epilepsy prior to starting the ketogenic diet (KD) and 3 to 4 weeks after KD therapy. Elevated beta hydroxybutyrate and cortisol levels were observed in all children on the KD. Free fatty acids increased 2.2-fold on the KD, with significant elevations in most polyunsaturated fatty acids (PUFA; arachidonate increased 1.6- to 2.9-fold and docosahexaenoate increased 1.5- to 4.0-fold). The rise in total serum arachidonate correlated with improved seizure control. Elevated PUFA may represent a key anticonvulsant mechanism of the KD. PMID- 12654977 TI - Facial hemangioma and cerebral corticovascular dysplasia: a syndrome associated with epilepsy. AB - The authors performed imaging studies in two children with epilepsy and congenital facial hemangioma. The first patient had dysplastic pericallosal arteries and frontal polymicrogyria. In the second patient, dysplastic arteries and dysplastic cortex lined the interhemispheric fissure, the dysplastic cortex bridging across the midline, which resulted in holoprosencephaly. Abnormal cortical development may underlie epilepsy in children with facial hemangioma. PMID- 12654978 TI - Periventricular heterotopia associated with chromosome 5p anomalies. AB - Periventricular heterotopia (PH) is characterized by neuronal nodules along the lateral ventricles. Whereas mutations in X-linked FLNA cause such cortical malformations, the authors report two cases of PH localizing to chromosome 5p. Both subjects have complex partial seizures. MRI demonstrated bilateral nodular PH, with subcortical heterotopia or focal gliosis. FISH identified a duplication of 5p15.1 [46,XX,dup(5)(p15.1p15.1)] and a trisomy of 5p15.33 [46,XY,der(14)t(5;14)(p15.33;p11.2) mat]. These findings suggest a new PH locus along the telomeric end of chromosome 5p. PMID- 12654980 TI - Cerebral blood flow changes induced by subthalamic stimulation in Parkinson's disease. AB - The authors used PET to show that the abnormal pattern of cerebral blood flow (CBF) activation described in PD is normalized by electrical stimulation of subthalamic nucleus. Improvement in bradykinesia correlated with increase in CBF due to stimulation in the supplementary motor area and anterior cingulate cortex. Compared to unilateral stimulation, bilateral stimulation induced a greater extent of activation in bilateral cortical areas and additional bilateral activation of globus pallidus. PMID- 12654979 TI - Neurotransmitter specificity of sympathetic denervation in Parkinson's disease. AB - In PD, orthostatic hypotension reflects sympathetic noradrenergic denervation. The authors assessed sympathetic cholinergic innervation by the quantitative sudomotor axon reflex test (QSART) in 12 patients who had sympathetic neurocirculatory failure, markedly decreased cardiac 6-[18F] fluorodopamine derived radioactivity, and subnormal plasma norepinephrine increments during standing. All 12 had normal QSART results. The sympathetic nervous system lesion in PD involves loss of postganglionic catecholaminergic but not cholinergic nerves. PMID- 12654981 TI - Atypical speech is rare in individuals with normal developmental histories. AB - The prevalence of atypical (right, bilateral) speech lateralization is unknown in normal populations. The authors investigated this by studying people with normal developmental histories but a later, specific adult neurologic event leading to intractable epilepsy. Fifty of 836 people receiving intracarotid amobarbital procedures (IAPs) met criteria of normal neurologic histories through age 15 years, with later head trauma or cerebral infection as probable cause of subsequent epilepsy. All 50 patients had left hemispheric speech on IAP. Atypical speech lateralization is rare unless there is also a positive neurologic history. PMID- 12654982 TI - Short-lasting impairment of tactile perception by 0.9Hz-rTMS of the sensorimotor cortex. AB - To test whether low-frequency repetitive transcranial magnetic stimulation (rTMS) of sensorimotor cortex (SM1) has prolonged effects on somatosensory function, eight subjects were given 900 TMS pulses over the left hand SM1 (0.9Hz, 90% of the resting motor threshold) or at sites 3 cm anterior or posterior to it. Tactile threshold of the right hand was increased for a short duration after rTMS over SM1, but two-point discrimination and median nerve SEPs were unaffected after rTMS at any sites. PMID- 12654983 TI - Clinical diagnosis of Fabry disease: whorl-like corneal opacity. PMID- 12654984 TI - Ipsilateral truncal sensory deficit in a patient with ophthalmic zoster sine herpete. PMID- 12654985 TI - Cluster-like headache time-locked to the menstrual cycle. PMID- 12654986 TI - Neuroleptic malignant-like syndrome due to donepezil and maprotiline. PMID- 12654987 TI - The role of the left frontal lobe in action naming: rTMS evidence. PMID- 12654988 TI - Opioids versus antidepressants in postherpetic neuralgia: a randomized, placebo controlled trial. PMID- 12654989 TI - Prevalence and clinical importance of sleep apnea in the first night after cerebral infarction. PMID- 12654990 TI - Isolated hand palsy due to cortical infarction: Localization of the motor hand area. PMID- 12654991 TI - Scope of neurological practice: different places, different models. PMID- 12654994 TI - Patient page. Memory problems after epilepsy surgery. PMID- 12654995 TI - A nomenclature for restriction enzymes, DNA methyltransferases, homing endonucleases and their genes. AB - A nomenclature is described for restriction endonucleases, DNA methyltransferases, homing endonucleases and related genes and gene products. It provides explicit categories for the many different Type II enzymes now identified and provides a system for naming the putative genes found by sequence analysis of microbial genomes. PMID- 12654996 TI - A survey of small RNA-encoding genes in Escherichia coli. AB - Small RNA (sRNA) molecules have gained much interest lately, as recent genome wide studies have shown that they are widespread in a variety of organisms. The relatively small family of 10 known sRNA-encoding genes in Escherichia coli has been significantly expanded during the past two years with the discovery of 45 novel genes. Most of these genes are still uncharacterized and their cellular roles are unknown. In this survey we examined the sequence and genomic features of the 55 currently known sRNA-encoding genes in E.coli, attempting to identify their common characteristics. Such characterization is important for both expanding our understanding of this unique gene family and for improving the methods to predict and identify sRNA-encoding genes based on genomic information. PMID- 12654998 TI - Scp160p associates with specific mRNAs in yeast. AB - Scp160p is a multiple KH-domain RNA-binding protein in yeast that has been demonstrated previously to associate with both soluble and membrane-bound polyribosomes as an mRNP component. One key question that has remained unanswered, however, is whether the mRNAs in these mRNP complexes are random or specific. We have addressed this question using microarray analyses of RNAs released from affinity isolated Scp160p-containing complexes, compared with total RNA controls from the same lysates. Our results, confirmed by quantitative RT-PCR analysis, clearly demonstrate that Scp160p associates with specific rather than with random messages, and that among the enriched targets are DHH1, YOR338W, BIK1, YOL155C and NAM8. Furthermore, loss of Scp160p resulted in a significant change in both the abundance and distribution between soluble and membrane associated fractions for at least one of these messages (YOR338W), and in a subtle yet significant shift from soluble polyribosomes to soluble mRNPs for at least two of these target messages (DHH1 and YOR338W). Together, these data not only identify specific mRNA targets associated with Scp160p in vivo, they demonstrate that the association of Scp160p with these messages is biologically relevant. PMID- 12654997 TI - A structural and primary sequence comparison of the viral RNA-dependent RNA polymerases. AB - A systematic bioinformatic approach to identifying the evolutionarily conserved regions of proteins has verified the universality of a newly described conserved motif in RNA-dependent RNA polymerases (motif F). In combination with structural comparisons, this approach has defined two regions that may be involved in unwinding double-stranded RNA (dsRNA) for transcription. One of these is the N terminal portion of motif F and the second is a large insertion in motif F present in the RNA-dependent RNA polymerases of some dsRNA viruses. PMID- 12654999 TI - DNA helix: the importance of being GC-rich. AB - A new explanation for the emergence of heavy (GC-rich) isochores is proposed, based on the study of thermostability, bendability, ability to B-Z transition and curvature of the DNA helix. The absolute values of thermostability, bendability and ability to B-Z transition correlated positively with GC content, whereas curvature correlated negatively. The relative values of these parameters were determined as compared to randomized sequences. In genes and intergenic spacers of warm-blooded animals, both the relative bendability and ability to B-Z transition increased with elevation of GC content, whereas the relative thermostability and curvature decreased. The usage of synonymous codons in GC rich genes was also found to augment bendability and ability to B-Z transition and to reduce thermostability of DNA (as compared to synonymous codons with the same GC content). The analysis of transposable elements (Alu and B2 repeats in the human and mouse) showed that the level of their divergence from the consensus sequence positively correlated with relative bendability and ability to B-Z transition and negatively with relative thermostability. The bendability and ability to B-Z transition are known to relate to open chromatin and active transcription, whereas curvature facilitates chromatin condensation. Because heavy isochores are known to be gene-rich and show a high level of transcription, it is suggested here that isochores arose not as an adaptation to elevated temperature but because of a certain grade of general organization and correspondingly advanced level of genomic organization, reflected in genome structuring, with physical properties of DNA in the gene-rich regions being optimized for active transcription and in the gene-poor regions for chromatin condensation ('transcription/grade' concept). PMID- 12655000 TI - Thyroid-specific transcription factors control Hex promoter activity. AB - The homeobox-containing gene Hex is expressed in several cell types, including thyroid follicular cells, in which it regulates the transcription of tissue- specific genes. In this study the regulation of Hex promoter activity was investigated. Using co- transfection experiments, we demonstrated that the transcriptional activity of the Hex gene promoter in rat thyroid FRTL-5 cells is approximately 10-fold greater than that observed in HeLa and NIH 3T3 cell lines (which do not normally express the Hex gene). To identify the molecular mechanisms underlying these differences, we evaluated the effect of the thyroid- specific transcription factor TTF-1 on the Hex promoter activity. TTF-1 produced 3-4-fold increases in the Hex promoter activity. Gel- retardation assays and mutagenesis experiments revealed the presence of functionally relevant TTF-1 binding sites in the Hex promoter region. These in vitro data may also have functional relevance in vivo, since a positive correlation between TTF-1 and Hex mRNAs was demonstrated in human thyroid tissues by means of RT-PCR analysis. The TTF-1 effect, however, is not sufficient to explain the difference in Hex promoter activity between FRTL-5 and cells that do not express the Hex gene. For this reason, we tested whether Hex protein is able to activate the Hex promoter. Indeed, co-transfection experiments indicate that Hex protein is able to increase the activity of its own promoter in HeLa cells approximately 4-fold. TTF-1 and Hex effects are additive: when transfected together in HeLa cells, the Hex promoter activity is increased 6-7-fold. Thus, the contemporary presence of both TTF-1 and Hex could be sufficient to explain the higher transcriptional activity of the Hex promoter in thyroid cells with respect to cell lines that do not express the Hex gene. These findings demonstrate the existence of direct cross regulation between thyroid-specific transcription factors. PMID- 12655002 TI - AINTEGUMENTA utilizes a mode of DNA recognition distinct from that used by proteins containing a single AP2 domain. AB - The Arabidopsis protein AINTEGUMENTA (ANT) is an important regulator of organ growth during flower development. ANT is a member of the AP2 subclass of the AP2/ERF family of plant-specific transcription factors. These proteins contain either one or two copies of a DNA-binding domain called the AP2 domain. Here, it is shown that ANT can act as a transcriptional activator in yeast through binding to a consensus ANT-binding site. This activity was used as the basis for a genetic screen to identify amino acids that are critical for the DNA binding ability of ANT. Mutants that showed reduced or no activation of a reporter gene under the control of ANT-binding sites were identified. The mutations identified in the screen as well as additional site-directed mutations suggest that the mode of DNA recognition by members of the AP2 subfamily is distinct from that of ERF proteins. Surprisingly, it appears that each AP2 domain of ANT uses different amino acids to contact DNA. Identification of several linker mutations argues that this sequence acts in the positioning of each AP2 domain on the DNA or makes direct DNA contacts. PMID- 12655001 TI - Conditional gene expression by controlling translation with tetracycline-binding aptamers. AB - We present a conditional gene expression system in Saccharomyces cerevisiae which exploits direct RNA-metabolite interactions as a mechanism of genetic control. We inserted preselected tetracycline (tc) binding aptamers into the 5'-UTR of a GFP encoding mRNA. While aptamer insertion generally reduces GFP expression, one group of aptamers displayed an additional, up to 6-fold, decrease in fluorescence upon tc addition. Regulation is observed for aptamers inserted cap-proximal or near the start codon, but is more pronounced from the latter position. Increasing the thermodynamic stability of the aptamer augments regulation but reduces expression of GFP. Decreasing the stability leads to the opposite effect. We defined nucleotides which influence the regulatory properties of the aptamer. Exchanging a nucleotide probably involved in tc binding only influences regulation, while mutations at another position alter expression in the absence of tc, without affecting regulation. Thus, we have developed and characterized a regulatory system which is easy to establish and controlled by a non-toxic, small ligand with good cell permeability. PMID- 12655003 TI - A non-redundant microarray of genes for two related bacteria. AB - A microarray with sequences from the annotated open reading frames (ORFs) in Salmonella enterica subspecies 1, serovar Typhimurium was supplemented with annotated chromosomal ORFs from serovar Typhi that are divergent from Typhimurium (>10% DNA sequence divergence). This non- redundant array was used to (i) measure changes in gene copy number in DNA from actively growing versus stationary Typhi and (ii) to reveal the transcriptional response of Typhi to peroxide, a stress similar to that experienced when they are phagocytosed by macrophages. In S.enterica subspecies 1, pairs of genomes differ in the presence or absence of approximately 10% of their genes. An array twice the size of that needed to cover all ORFs for one genome could carry close homologs of all the ORFs for 10 genomes. Non-redundant DNA arrays could be constructed for any group of closely related organisms that differ by the presence and absence of a few genes. PMID- 12655004 TI - The human Imp3 and Imp4 proteins form a ternary complex with hMpp10, which only interacts with the U3 snoRNA in 60-80S ribonucleoprotein complexes. AB - Ribosome biogenesis requires a vast number of trans-acting factors many of which are required for the chemical modification and processing of the pre-rRNA component. The U3 snoRNP complex is required for the early cleavage steps in pre rRNA processing. We have cloned cDNAs encoding the human and mouse homologs of the yeast U3 snoRNP-associated proteins Imp3 and Imp4. Both human proteins localize to nucleoli and interact with the U3 snoRNA. The results of complementation experiments show that, in contrast to mouse Imp4, mouse Imp3 can partially alleviate the growth defect of the corresponding yeast null strain, indicating that the role of Imp3 in pre-rRNA processing is evolutionarily conserved. The results of density gradient centrifugation experiments show that, in contrast to hU3-55K, the human Imp3 and Imp4 proteins predominantly interact with the U3 snoRNA in 60-80S ribonucleoprotein complexes. In addition, we have found that hImp3, hImp4 and hMpp10 can form a stable hetero-trimeric complex in vitro, which is generated by direct interactions of both hImp3 and hImp4 with hMpp10. The analysis of hImp3 and hImp4 mutants indicated that their binding to hMpp10 correlates with their nucleolar accumulation, strongly suggesting that the formation of the ternary complex of hImp3, hImp4 and hMpp10 is required for their association with nucleolar components. PMID- 12655005 TI - Functional cooperation between exonucleases and endonucleases--basis for the evolution of restriction enzymes. AB - Many types of restriction enzymes cleave DNA away from their recognition site. Using the type III restriction enzyme, EcoP15I, which cleaves DNA 25-27 bp away from its recognition site, we provide evidence to show that an intact recognition site on the cleaved DNA sequesters the restriction enzyme and decreases the effective concentration of the enzyme. EcoP15I restriction enzyme is shown here to perform only a single round of DNA cleavage. Significantly, we show that an exonuclease activity is essential for EcoP15I restriction enzyme to perform multiple rounds of DNA cleavage. This observation may hold true for all restriction enzymes cleaving DNA sufficiently far away from their recognition site. Our results highlight the importance of functional cooperation in the modulation of enzyme activity. Based on results presented here and other data on well-characterised restriction enzymes, a functional evolutionary hierarchy of restriction enzymes is discussed. PMID- 12655006 TI - Influence of DNA torsional rigidity on excision of 7,8-dihydro-8-oxo-2' deoxyguanosine in the presence of opposing abasic sites by human OGG1 protein. AB - The human protein OGG1 (hOGG1) targets the highly mutagenic base 7,8-dihydro-8 oxo-2'-deoxyguanosine (8-oxodG) and shows a high specificity for the opposite DNA base. Abasic sites can arise in DNA in close opposition to 8-oxodG either during repair of mismatched bases (i.e. 8-oxodG/A mismatches) or, more frequently, as a consequence of ionizing radiation exposure. Bistranded DNA lesions may remain unrepaired and lead to cell death via double-strand break formation. In order to explore the role of damaged-DNA dynamics in recognition/excision by the hOGG1 repair protein, specific oligonucleotides containing an 8-oxodG opposite an abasic site, at different relative distances on the complementary strand, were synthesized. Rotational dynamics were studied by means of fluorescence polarization anisotropy decay experiments and the torsional elastic constant as well as the hydrodynamic radius of the DNA fragments were evaluated. Efficiency of excision of 8-oxodG was tested using purified human glycosylase. A close relation between the twisting flexibility of the DNA fragment and the excision efficiency of the oxidative damage by hOGG1 protein within a cluster was found. PMID- 12655007 TI - Pathology-related substitutions in human mitochondrial tRNA(Ile) reduce precursor 3' end processing efficiency in vitro. AB - The human mitochondrial genome encodes 22 tRNAs interspersed among the two rRNAs and 11 mRNAs, often without spacers, suggesting that tRNAs must be efficiently excised. Numerous maternally transmitted diseases and syndromes arise from mutations in mitochondrial tRNAs, likely due to defect(s) in tRNA metabolism. We have systematically explored the effect of pathogenic mutations on tRNA(Ile) precursor 3' end maturation in vitro by 3'-tRNase. Strikingly, four pathogenic tRNA(Ile) mutations reduce 3'-tRNase processing efficiency (V(max) / K(M)) to approximately 10-fold below that of wild-type, principally due to lower V(max). The structural impact of mutations was sought by secondary structure probing and wild-type tRNA(Ile) precursor was found to fold into a canonical cloverleaf. Among the mutant tRNA(Ile) precursors with the greatest 3' end processing deficiencies, only G4309A displays a secondary structure substantially different from wild-type, with changes in the T domain proximal to the substitution. Reduced efficiency of tRNA(Ile) precursor 3' end processing, in one case associated with structural perturbations, could thus contribute to human mitochondrial diseases caused by mutant tRNAs. PMID- 12655008 TI - Repair of hydrolytic DNA deamination damage in thermophilic bacteria: cloning and characterization of a Vsr endonuclease homolog from Bacillus stearothermophilus. AB - Hydrolytic deamination of 5-methyl cytosine in double stranded DNA results in formation of a T/G mismatch that-if left unrepaired-leads to a C-->T transition mutation in half of the progeny. In addition to several mismatch-specific glycosylases that have been found in both pro- and eukaryotes to channel this lesion into base excision repair by removing the T from the mismatch, Vsr endonuclease from Escherichia coli has been described which initiates repair by an endonucleolytic strand incision 5' to the mismatched T. We have isolated a gene coding for a homolog of E.coli Vsr endonuclease from the thermophilic bacterium Bacillus stearothermophilus H3 (Vsr.Bst) using a method that allows PCR amplification with degenerated primers of gene segments which code for only one highly conserved amino acid region. Vsr.Bst was produced heterologously in E.coli and purified to apparent homogeneity. Vsr.Bst specifically incises heteroduplex DNA with a preference for T/G mismatches. The selectivity of Vsr.Bst for the sequence context of the T/G mismatch appears less pronounced than for Vsr.Eco. PMID- 12655009 TI - Molecular recognition properties of IGS-mediated reactions catalyzed by a Pneumocystis carinii group I intron. AB - We report the development, analysis and use of a new combinatorial approach to analyze the substrate sequence dependence of the suicide inhibition, cyclization, and reverse cyclization reactions catalyzed by a group I intron from the opportunistic pathogen Pneumocystis carinii. We demonstrate that the sequence specificity of these Internal Guide Sequence (IGS)-mediated reactions is not high. In addition, the sequence specificity of suicide inhibition decreases with increasing MgCl(2) concentration, reverse cyclization is substantially more sequence specific than suicide inhibition, and multiple reverse cyclization products occur, in part due to the formation of multiple cyclization intermediates. Thermodynamic analysis reveals that a base pair at position -4 of the resultant 5' exon-IGS (P1) helix is crucial for tertiary docking of the P1 helix into the catalytic core of the ribozyme in the suicide inhibition reaction. In contrast to results reported with a Tetrahymena ribozyme, altering the sequence of the IGS of the P.carinii ribozyme can result in a marked reduction in tertiary stability of docking the resultant P1 helix into the catalytic core of the ribozyme. Finally, results indicate that RNA targeting strategies which exploit tertiary interactions could have low specificity due to the tolerance of mismatched base pairs. PMID- 12655010 TI - Inhibition of HCV NS3 protease by RNA aptamers in cells. AB - Non-structural protein 3 (NS3) of hepatitis C virus (HCV) has two distinct activities, protease and helicase, which are essential for HCV proliferation. In previous work, we obtained RNA aptamers (G9-I, II and III) which specifically bound the NS3 protease domain (DeltaNS3), efficiently inhibiting protease activity in vitro. To utilize these aptamers in vivo, we constructed a G9 aptamer expression system in cultured cells, using the cytomegarovirus enhancer + chicken beta-actin globin (CAG) promoter. By conjugating the cis-acting genomic human hepatitis delta virus (HDV) ribozyme and G9-II aptamer, a chimeric HDV ribozyme G9-II aptamer (HA) was constructed, which was used to produce stable RNA in vivo and to create tandem repeats of the functional unit. To target the transcribed RNA aptamers to the cytoplasm, the minimal mutant of constitutive transport element (CTE), derived from type D retroviruses, was conjugated at the 3' end of HA (HAC). Transcript RNAs from (HA)(n) and (HAC)(n) were processed into the G9-II aptamer unit by the cis-acting HDV ribozyme, both in vitro and in vivo. Efficient protease inhibition activity of HDV ribozyme-G9-II aptamer expression plasmid was demonstrated in HeLa cells. Protease inhibition activity level of tandem chimeric aptamers, (HA)(n) and (HAC)(n), rose with the increase of n from 1 to 4. PMID- 12655013 TI - Absolute mRNA concentrations from sequence-specific calibration of oligonucleotide arrays. AB - Oligonucleotide microarrays are based on the hybridization of labeled mRNA molecules to short length oligonucleotide probes on a glass surface. Two effects have been shown to affect the raw data: the sequence dependence of the probe hybridization properties and the chemical saturation resulting from surface adsorption processes. We address both issues simultaneously using a physically motivated hybridization model. Based on publicly available calibration data sets, we show that Langmuir adsorption accurately describes GeneChip hybridization, with model parameters that we predict from the sequence composition of the probes. Because these parameters have physical units, we are able to estimate absolute mRNA concentrations in picomolar. Additionally, by accounting for chemical saturation, we substantially reduce the compressive bias of differential expression estimates that normally occurs toward high concentrations. PMID- 12655012 TI - Specific SR protein-dependent splicing substrates identified through genomic SELEX. AB - The Drosophila pre-mRNA splicing factor B52 (SRp55) is essential for fly development, but splicing of RNAs of specific genes tested previously is normal in B52-null animals, presumably due to partial functional redundancy with other SR proteins. To identify B52-dependent splicing substrates in vivo, we selected genomic sequence fragments whose transcripts bind B52. Almost all of the corresponding genes having a known function encode either transcription factors or components of signal transduction pathways, with the B52- binding fragments located to not only exonic but also intronic regions. Some pre-mRNAs from these genes showed splicing defects in the B52-null mutant. These results indicate that B52 has unique functions in the removal of some introns during development, and plays a critical role in cellular regulatory networks. PMID- 12655014 TI - Transcription of genes encoding trans-acting factors required for rRNA maturation/ribosomal subunit assembly is coordinately regulated with ribosomal protein genes and involves Rap1 in Saccharomyces cerevisiae. AB - We demonstrate that the genes encoding trans- acting factors essential for pre rRNA processing/ribosomal subunit assembly are responsive to various kinds of stresses such as heat shock, nitrogen deprivation and a secretory defect, in coordination with ribosomal protein genes in Saccharomyces cerevisiae. The rap1 17 mutation, which produces the C-terminally truncated protein of a transcriptional factor Rap1p, affects transcriptional repression of the trans acting factor genes due to a secretory defect as shown previously for both ribosomal protein and rRNA genes. PMID- 12655011 TI - What's in the genome of a filamentous fungus? Analysis of the Neurospora genome sequence. AB - The German Neurospora Genome Project has assembled sequences from ordered cosmid and BAC clones of linkage groups II and V of the genome of Neurospora crassa in 13 and 12 contigs, respectively. Including additional sequences located on other linkage groups a total of 12 Mb were subjected to a manual gene extraction and annotation process. The genome comprises a small number of repetitive elements, a low degree of segmental duplications and very few paralogous genes. The analysis of the 3218 identified open reading frames provides a first overview of the protein equipment of a filamentous fungus. Significantly, N.crassa possesses a large variety of metabolic enzymes including a substantial number of enzymes involved in the degradation of complex substrates as well as secondary metabolism. While several of these enzymes are specific for filamentous fungi many are shared exclusively with prokaryotes. PMID- 12655015 TI - Conserved sequence elements associated with exon skipping. AB - One of the major forms of alternative splicing, which generates multiple mRNA isoforms differing in the precise combinations of their exon sequences, is exon skipping. While in constitutive splicing all exons are included, in the skipped pattern(s) one or more exons are skipped. The regulation of this process is still not well understood; so far, cis- regulatory elements (such as exonic splicing enhancers) were identified in individual cases. We therefore set to investigate the possibility that exon skipping is controlled by sequences in the adjacent introns. We employed a computer analysis on 54 sequences documented as undergoing exon skipping, and identified two motifs both in the upstream and downstream introns of the skipped exons. One motif is highly enriched in pyrimidines (mostly C residues), and the other motif is highly enriched in purines (mostly G residues). The two motifs differ from the known cis-elements present at the 5' and 3' splice site. Interestingly, the two motifs are complementary, and their relative positional order is conserved in the flanking introns. These suggest that base pairing interactions can underlie a mechanism that involves secondary structure to regulate exon skipping. Remarkably, the two motifs are conserved in mouse orthologous genes that undergo exon skipping. PMID- 12655017 TI - An approach to identify over-represented cis-elements in related sequences. AB - Computational identification of transcription factor binding sites is an important research area of computational biology. Positional weight matrix (PWM) is a model to describe the sequence pattern of binding sites. Usually, transcription factor binding sites prediction methods based on PWMs require user defined thresholds. The arbitrary threshold and also the relatively low specificity of the algorithm prevent the result of such an analysis from being properly interpreted. In this study, a method was developed to identify over represented cis-elements with PWM-based similarity scores. Three sets of closely related promoters were analyzed, and only over- represented motifs with high PWM similarity scores were reported. The thresholds to evaluate the similarity scores to the PWMs of putative transcription factors binding sites can also be automatically determined during the analysis, which can also be used in further research with the same PWMs. The online program is available on the website: http://www.bioinfo.tsinghua.edu.cn/- zhengjsh/OTFBS/. PMID- 12655016 TI - 3'-Exonuclease resistance of DNA oligodeoxynucleotides containing O6-[4-oxo-4-(3 pyridyl)butyl]guanine. AB - Tobacco-specific nitrosamine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), is a chemical carcinogen thought to be involved in the initiation of lung cancer in smokers. NNK is metabolically activated to methylating and pyridyloxobutylating species that form promutagenic adducts with DNA nucleobases, e.g. O(6)-[4-oxo-4-(3-pyridyl)butyl]guanine (O(6)-POB-dG). O(6)-POB-dG is a strongly mispairing DNA lesion capable of inducing both G-->A and G-->T base changes, suggesting its importance in NNK mutagenesis and carcinogenesis. Our earlier investigations have identified the ability of O(6)-POB-dG to hinder DNA digestion by snake venom phosphodiesterase (SVPDE), a 3'-exonuclease commonly used for DNA ladder sequencing and as a model enzyme to test nuclease sensitivity of anti-sense oligonucleotide drugs. We now extend our investigation to three other enzymes possessing 3'-exonuclease activity: bacteriophage T4 DNA polymerase, Escherichia coli DNA polymerase I, and E.coli exonuclease III. Our results indicate that, unlike SVPDE, 3'-exonuclease activities of these three enzymes are not blocked by O(6)-POB-dG lesion. Conformational analysis and molecular dynamics simulations of DNA containing O(6)-POB-dG suggest that the observed resistance of the O(6)-POB-dG lesion to SVPDE-catalyzed hydrolysis may result from the structural changes in the DNA strand induced by the O(6)-POB group, including C3'-endo sugar puckering and the loss of stacking interaction between the pyridyloxobutylated guanine and its flanking bases. In contrast, O(6) methylguanine lesion used as a control does not induce similar structural changes in DNA and does not prevent its digestion by SVPDE. PMID- 12655018 TI - Analysis of the conformational energy landscape of human snRNA with a metric based on tree representation of RNA structures. AB - It is an outstanding problem to clarify how the RNA sequence is related to its structure and biological functions. We developed a simplified definition of a metric for tree representation of RNA secondary structures and analyzed the conformational energy landscapes of human spliceosomal snRNAs. We discuss the structural properties of the biological sequence by calculating the conformational energy landscapes based on the structural distance between each of the pairs in the set of suboptimal structures. The new index value is introduced for estimating the shapes of distribution patterns in conformational energy landscapes. We apply our method to the five human snRNAs and show that U1 snRNA has a multi-valley profile of the landscape, whereas the landscapes of the other four snRNAs have one steep valley. This result reflects different biological functions of these snRNAs in the pre-mRNA splicing process. The results of analyzing tRNAs and rRNAs show that the conformational energy landscapes of these sequences have multi-valley profiles. PMID- 12655020 TI - Bypassing antibiotic selection: positive screening of genetically modified cells with an antigen-dependent proliferation switch. AB - While antibiotic selection has been routinely used for the selection of genetically modified cells, administration of cytotoxic drugs often leads to deleterious effects not only to inert cells but also to transfected or transduced ones. In this study, we propose an Antigen-MEdiated Genetically modified cell Amplification (AMEGA) system employing antibody/receptor chimeras without antibiotic selection. Based on a rational design where the extracellular domains of dimeric erythropoietin receptor (EpoR) or gp130 were substituted with heterodimeric V(H)/V(L) regions of anti-hen egg lysozyme (HEL) antibody and EpoR D2 domains, the genes encoding the chimeras as well as a model transgene, enhanced green fluorescent protein (EGFP), were retrovirally infected into IL-3 dependent Ba/F3 cells followed by direct HEL selection in the absence of IL-3. After a single round of selection, EGFP-positive cells were selectively amplified, resulting in a population of almost 100% positive cells. The AMEGA without antibiotic selection will not harm normal cells, which will be especially useful for increasing the efficacy for stem cell-based gene therapy. PMID- 12655021 TI - Amplification of repeat-containing transcribed sequences (ARTS): a transcriptome fingerprinting strategy to detect functionally relevant microsatellite mutations in cancer. AB - Cancer is a genetic disease caused by mutations in somatic cells. Those that carry advantageous mutations are favoured by natural selection. In most cancers, genetic instability increases mutation rate and facilitates cancer cell evolution. Microsatellite instability (MSI), due to defects of the DNA mismatch repair system, affects in particular repeat sequences (microsatellites) scattered throughout the genome. As mutations in expressed genes are more likely to be functional, we developed a procedure for the systematic identification of mutant repeat-containing expressed sequences (amplification of repeat-containing transcribed sequences, ARTS). The entire cell mRNA was converted into short double-stranded cDNA fragments linked to an adapter at both ends. Repeat containing cDNA fragments were PCR amplified using the adapter-specific primer in combination with different arbitrary primers including the repeat. ARTS yielded discrete PCR products with lengths that were directly correlated to the lengths of the endogenous repeats. Comparison between ARTS products obtained from control cells and cancer cells with microsatellite instability (MSI+) revealed mRNAs carrying insertions or deletions at repeats. The subsequent sequencing allowed the identification of a series of frameshift-mutated mRNAs in MSI+ cancer cells, including the already described mutant BAX transcript. These data show that ARTS provides an unbiased genome-wide approach to the discovery of functionally relevant genes that could be affected by MSI in cancer. PMID- 12655019 TI - Self-association and conformational properties of RAG1: implications for formation of the V(D)J recombinase. AB - RAG1 and RAG2 catalyze the initial DNA cleavage steps in V(D)J recombination. Fundamental properties of these proteins remain largely unknown. Here, self association and conformational properties of murine core RAG1 (residues 384-1008) were examined. As determined by multi-angle laser light scattering measurements, the molecular masses of two predominant core RAG1 species corresponded to dimeric and tetrameric states. Similar results were obtained using a RAG1 fragment containing residues 265-1008, indicating that a non-core portion of RAG1 does not alter the oligomerization states observed for the core region. The fraction of core RAG1 in the tetrameric state increased significantly at lower ionic strengths (0.2 versus 0.5 M NaCl), indicating that this oligomeric form may factor into the physiological function of RAG1. In addition, the secondary structural content of core RAG1, obtained by circular dichroism spectroscopy, demonstrated a significant dependence on ionic strength with a 26% increase in alpha-helical content from 0.2 to 1.0 M NaCl. Together, these results indicate that structural and oligomerization properties of core RAG1 are strongly dependent on electrostatic interactions. Furthermore, the secondary structure of core RAG1 changes upon binding to DNA, with larger increases in alpha-helical content upon binding to the recombination signal sequence (RSS) as compared with non-sequence-specific DNA. As shown by electrophoretic mobility shift assays, higher order oligomeric forms of core RAG1 bound to the canonical RSS. Furthermore, core RAG2 (residues 1-387) formed complexes with multimeric RAG1 species bound to a single RSS, providing additional support for the physiological relevance of higher order oligomeric states of RAG1. PMID- 12655022 TI - Internal 32P-labeling of L-deoxyoligonucleotides. AB - A general two step procedure for the internal labeling of L deoxyoligonucleotides, Spiegelmers, has been developed. Through radioactive labeling oligonucleotides can easily be detected and monitored in biological samples. T4 polynucleotide kinase is shown to efficiently phosphorylate strands of L-nucleic acids which allows the labeling with phosphorous isotopes such as (32)P. In order to protect the terminal phosphate label against unspecific phosphatases, one of two fragments of a Spiegelmer is enzymatically phosphorylated with [gamma-(32)P]ATP. In a second step we used a template- directed chemical ligation reaction in order to attach the labeled oligonucleotide to the other fragment to yield the full-length Spiegelmer with an internal [(32)P]phosphodiester bond. It has been shown that the functionality of a chemically ligated Spiegelmer is still retained. PMID- 12655023 TI - Light-directed 5'-->3' synthesis of complex oligonucleotide microarrays. AB - Light-directed synthesis of high-density microarrays is currently performed in the 3'-->5' direction due to constraints in existing synthesis chemistry. This results in the probes being unavailable for many common types of enzymatic modification. Arrays that are synthesized in the 5'-->3' direction could be utilized to perform parallel genotyping and resequencing directly on the array surface, dramatically increasing the throughput and reducing the cost relative to existing techniques. In this report we demonstrate the use of photoprotected phosphoramidite monomers for light-directed array synthesis in the 5'-->3' direction, using maskless array synthesis technology. These arrays have a dynamic range of >2.5 orders of magnitude, sensitivity below 1 pM and a coefficient of variance of <10% across the array surface. Arrays containing >150,000 probe sequences were hybridized to labeled mouse cRNA producing highly concordant data (average R(2) = 0.998). We have also shown that the 3' ends of array probes are available for sequence-specific primer extension and ligation reactions. PMID- 12655024 TI - Data extraction from composite oligonucleotide microarrays. AB - Microarray or DNA chip technology is revolutionizing biology by empowering researchers in the collection of broad-scope gene information. It is well known that microarray-based measurements exhibit a substantial amount of variability due to a number of possible sources, ranging from hybridization conditions to image capture and analysis. In order to make reliable inferences and carry out quantitative analysis with microarray data, it is generally advisable to have more than one measurement of each gene. The availability of both between-array and within-array replicate measurements is essential for this purpose. Although statistical considerations call for increasing the number of replicates of both types, the latter is particularly challenging in practice due to a number of limiting factors, especially for in-house spotting facilities. We propose a novel approach to design so-called composite microarrays, which allow more replicates to be obtained without increasing the number of printed spots. PMID- 12655025 TI - RNaseCut: a MALDI mass spectrometry-based method for SNP discovery. AB - MALDI mass spectrometry is an established platform for high-throughput genotyping of single nucleotide polymorphisms (SNPs). For many species and also for specific ethnic groups, the number of described SNPs is far from sufficient. Here we present a method for SNP discovery that can use existing MALDI genotyping platforms and is automation-compatible. The method is based on in vitro RNA transcripts from PCR products, that can be used to obtain highly informative sequence fingerprints by digestion with the guanosine- specific ribonuclease T1. In these fingerprints, a mutation can be detected as either a mass shift, absence of an existing peak or appearance of an additional peak. Due to mass-degeneracy of fragments and multiple presence of shorter fragments in a given sequence, a certain fraction of possible mutations will remain undetected with this method. Screening of both strands from one PCR product is possible by using T3- and T7 tailed primers and the respective RNA polymerases, and markedly decreases the probability of missing an existing SNP. The use of mass-shifted nucleotides can significantly reduce fragment overlaps and hence increase detectability. We have used a simulation of RNase digests of a set of randomly generated sequences to provide estimates for the general detection probability in dependence on PCR product length. A software package is provided that helps to design PCR primers by plotting out regions with a high SNP discovery score, calculates expected mass fingerprints and peaklists from the target sequence selected for screening and helps in interpretation of digest spectra. PMID- 12655026 TI - A simple and cost-effective method for producing small interfering RNAs with high efficacy. AB - Small interfering RNAs (siRNAs) are powerful RNA interference (RNAi) reagents for directed post- transcriptional gene silencing. Exogenous siRNA is frequently used in RNAi studies. However, due to profound differences in the activity of siRNAs targeted to different regions of a gene, several reagents may have to be screened for optimal activity. This approach is expensive due to the cost of chemical synthesis of RNAs. We report a technically simple, quick and cost-effective method for the production of siRNAs that makes use of in vitro transcription and deoxyribozyme digestion of the transcripts to produce the desired sequence and length. The method allows for several siRNAs to be produced in parallel at much reduced costs. The siRNAs produced with this method were tested in MDA-MB-231 human breast cancer cells for efficacy against the type 1 insulin-like growth factor receptor (IGF1R) mRNA and they caused dose-dependent inhibition of IGF1R expression comparable to that induced by chemically synthesised siRNAs of the same sequence. This method is also useful for producing long RNA fragments of defined length and sequence that may be difficult to synthesise chemically, and also for producing large quantities of RNAs for applications including structural studies and the study of interactions between RNA and other molecules, such as proteins, other nucleic acids and drugs. PMID- 12655028 TI - Construction and purification of site-specifically modified DNA templates for transcription assays. AB - Chemical and physical agents can alter the structure of DNA by modifying the bases and the phosphate-sugar backbone, consequently compromising both replication and transcription. During transcription elongation, RNA polymerase complexes can stall at a damaged site in DNA and mark the lesion for repair by a subset of proteins that are utilized to execute nucleotide excision repair, a pathway commonly associated with the removal of bulky DNA damage from the genome. This RNA polymerase-induced repair pathway is called transcription-coupled nucleotide excision repair. Although our understanding of DNA lesion effects on transcription elongation and the associated effects of stalled transcription complexes on DNA repair is broadening, the attainment of critical data is somewhat impeded by labor-intensive, time- consuming processes that are required to prepare damaged DNA templates. Here, we describe an approach for building linear DNA templates that contain a single, site-specific DNA lesion and support transcription by human RNA polymerase II. The method is rapid, making use of biotin-avidin interactions and paramagnetic particles to purify the final product. Data are supplied demonstrating that these templates support transcription, and we emphasize the potential versatility of the protocol and compare it with other published methods. PMID- 12655027 TI - Mechanisms underlying the impact of humic acids on DNA quantification by SYBR Green I and consequences for the analysis of soils and aquatic sediments. AB - DNA quantification of soils and sediments is useful for the investigation of microbial communities and for the acquisition of their genomes that are exploited for the production of natural products. However, in such samples DNA quantification is impaired by humic acids (HA). Due to its lack of specificity and sensitivity, UV spectrophotometry cannot be applied. Consequently, fluorimetric assays applying Hoechst (H) 33258 or PicoGreen (PG) are used. Here, we investigated the SYBR Green I (SG) assay, which was also affected by HA, but was found to be 25- and 1.7-fold more sensitive compared to the H 33258 and PG assays, respectively. Spectrophotometric, fluorimetric and quenching studies as well as gel mobility shift assays suggested that the effect of HA on the SG assay was based on an inner filter effect, collisional quenching and binding of SG to HA. As to the latter finding, the standard 6250-fold dilution of the SG reagent was optimised to a 2000-fold dilution. Although the sensitivity of the optimised SG assay was reduced by a factor of 1.3, the interfering effect of HA could be reduced up to 22-fold. A significant reduction of HA interferences by lowering the pH of the assay was not observed. Finally, the performance of the modified SG assay and the corresponding evaluation methods were verified by the determination of DNA recoveries and concentrations of standards and environmental samples in comparison to the PG assay. PMID- 12655029 TI - Mechanisms of increased liver tissue repair and survival in diet-restricted rats treated with equitoxic doses of thioacetamide. AB - Moderate dietary or caloric restriction (DR) modulates animal physiology in a beneficial fashion. Previously, we have reported an equitoxic dose experiment where liver injury in DR male Sprague-Dawley rats exposed to a low dose of thioacetamide (TA, 50 mg/kg) was similar to that observed in ad libitum fed (AL) rats exposed to a 12-fold higher dose (600 mg/kg). Paradoxically, the AL rats experienced 90% mortality while all of the DR rats, with the same amount of initial bioactivation-mediated liver injury, survived. The protection observed in the DR rats was due to efficient compensatory liver tissue repair, which was delayed and attenuated in the AL rats, leading to progression of liver injury. The objective of the present study was to investigate the molecular mechanisms of the enhanced tissue repair in the DR rats upon equitoxic challenge with TA. Promitogenic mechanisms and mediators such as proinflammatory cytokines (TNF alpha and IL-6), growth factors (TGF-alpha and HGF), and inducible nitric oxide synthase (iNOS) were estimated over a time course after equitoxic challenge (50 mg/kg to DR vs. 600 mg/kg to AL rats). Except for TNF-alpha, all other molecules were expressed earlier and in greater amount in the DR rats. IL-6 was 10-fold greater and peaked 12 h earlier; HGF also peaked 12 h sooner in the DR rats, when it was 2.5-fold greater than the value in the AL rats. TGF-alpha expression in livers of DR rats increased after TA administration and peaked at 24 h. In the AL rats, it was lower and peaked at 36 h. Diet restriction alone induced iNOS 2-fold in the DR rats and remained elevated until 12 h after TA administration, then declined thereafter. The lower iNOS activity in the AL rats further decreased after TA injection. DR rats exhibited higher apoptosis after thioacetamide administration, which further increased the efficiency of tissue repair. Taken together, these data indicate that even though the liver injury is near equal in AL and DR rats, sluggish signal transduction leads to delayed liver regeneration, progression of liver injury, and death in the AL rats. The equitoxic dose experiment indicates that stimulation of tissue repair is independent of the extent of initial liver injury and is governed by physiology of diet restriction. DR stimulates promitogenic signaling leading to a quick and timely response upon liver injury, arrest of progressive injury on one hand, and recovery from injury on the other, paving the way for survival of the DR rats. PMID- 12655030 TI - Expression of aryl hydrocarbon receptor repressor in normal human tissues and inducibility by polycyclic aromatic hydrocarbons in human tumor-derived cell lines. AB - Aryl hydrocarbon receptor repressor (AhRR) has been recently identified as a negative factor that suppresses aryl hydrocarbon receptor (AhR)-mediated transcriptional gene expression. In the present study, the expression level of AhRR in normal human tissues was determined. AhRR mRNA was detected in liver, breast, colon, kidney, lung, bladder, uterus, testis, ovary, and adrenal gland. The expression level in the testis was prominently high. AhRR mRNA was also detected in various human tissue-derived cell lines, HepG2 (hepatocellular carcinoma), MCF-7 (breast carcinoma), LS-180 (colon carcinoma), ACHN (renal carcinoma), A549 (lung carcinoma), HT-1197 (bladder carcinoma), HeLa (cervix of uterus adenocarcinoma), NEC14 (testis embryonal carcinoma), and OMC-3 (ovarian carcinoma). Since the expression level of AhRR mRNA was prominently high in HeLa cells, it is suggested that the high expression level of AhRR might work as a negative factor for the low inducibility of the CYP1 family in HeLa cells. The expression of AhRR mRNA was induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) or 3-methylchoranthrene (3-MC) in HepG2, MCF-7, LS-180, and OMC-3 cells, but not in ACHN, A549, HT-1197, HeLa, and NEC14 cells. The responsiveness was similar to the cell-specific inducibility of the CYP1 family. The inducibility of AhRR mRNA by nitropolycyclic aromatic hydrocarbons (NPAHs) as well as their parent PAHs was compared in HepG2 and OMC-3 cells. The chemical-specific inducibility of AhRR was also similar to that of the CYP1 family determined in our previous study. These results indicated that AhRR is also induced in chemical- and cell-specific manners. PMID- 12655031 TI - Gestational exposure to persistent organic pollutants: maternal liver residues, pregnancy outcome, and effects on hepatic gene expression profiles in the dam and fetus. AB - Dietary exposure of Inuit people to a mixture of pesticides and polychlorinated biphenyls, or persistent organic pollutants (POPs), during pregnancy is a public health concern. We examined the consequences of administering the mixture of 28 POPs found in the Inuit diet (at doses representing 10-1000 times dietary levels) by gavage to pregnant Sprague-Dawley rats either during gestation days 0-19 or 8 19. The levels of individual components of the POPs mixture in the maternal liver were measured by high-resolution mass spectrometry. On gestation day 20, dams were sacrificed and pregnancy outcome determined. RNA isolated from maternal and fetal livers was 32P-labeled for gene expression profiling. The concentrations of individual POPs were increased in maternal livers of dams gavaged with the 1000x POPs mixture by 10- to 500-fold. While exposure to POPs had no significant effects on pregnancy outcome, dramatic changes were observed in the gene expression profiles of both the maternal and fetal livers. The gene expression profiles of maternal and fetal female and male liver were distinct with respect to the numbers of transcripts detected, the genes expressed exclusively in control or POPs-exposed livers, and those for which expression was up- or downregulated. While different genes were affected in each group, the overall consequence of POPs exposure on hepatic gene expression profiles was to decrease both the numbers of genes expressed and the relative intensity of expression. Thus, in utero exposure to POPs alters hepatic gene expression in the dam and the fetus; these changes may have functional implications. PMID- 12655033 TI - Ultrafine carbon black particles enhance respiratory syncytial virus-induced airway reactivity, pulmonary inflammation, and chemokine expression. AB - Exposure to particulate matter (PM) may exacerbate preexisting respiratory diseases such as asthma, chronic obstructive pulmonary disease (COPD), bronchitis, and pneumonia. However, few experimental studies have addressed the effects of PM on lower respiratory tract (LRT) viral infection. Respiratory syncytial virus (RSV) is a major etiological agent for LRT infections in infants, the elderly, and the immunocompromised and may lead to chronic wheezing and the development of asthma in children. In this study, we examined the effects of carbon black (CB) on RSV-induced pulmonary inflammation, chemokine and cytokine expression, and airway hyperresponsiveness in a mouse model of RSV. Female BALB/c mice were instilled via the trachea (i.t.) with 1 x 106 plaque forming units (pfu) RSV or with uninfected culture media. On day 3 of infection, mice were i.t. instilled with either 40 micro g ultrafine CB particles or with saline. End points were examined on days 4, 5, 7, and 14 of RSV infection. Viral titer and clearance in the lung were unaffected by CB exposure. Neutrophil numbers were elevated on days 4 and 7, and lymphocyte numbers were higher on days 4 and 14 of infection in CB-exposed, RSV-infected mice. CB exposure also enhanced RSV-induced airway hyperresponsiveness to methacholine, bronchoalveolar lavage (BAL) total protein, and virus-associated chemokines monocyte chemoattractant protein (MCP 1), macrophage inflammatory protein (MIP-1 alpha), and regulated upon activation, normal T cell expressed and secreted (RANTES). MIP-1 alpha mRNA expression was increased in the alveolar epithelium, where ultrafine particles deposit in the lung. These data demonstrate a synergistic effect of ultrafine CB particles on RSV infection, and suggest a potential mechanism for increased respiratory infections in human populations after PM exposure. PMID- 12655032 TI - Male mice deficient in microsomal epoxide hydrolase are not susceptible to benzene-induced toxicity. AB - Enzymes involved in benzene metabolism are likely genetic determinants of benzene induced toxicity. Polymorphisms in human microsomal epoxide hydrolase (mEH) are associated with an increased risk of developing leukemia, specifically those associated with benzene. This study was designed to investigate the importance of mEH in benzene-induced toxicity. Male and female mEH-deficient (mEH-/-) mice and background mice (129/Sv) were exposed to inhaled benzene (0, 10, 50, or 100 ppm) 5 days/week, 6 h/day, for a two-week duration. Total white blood cell counts and bone marrow cell counts were used to assess hematotoxicity and myelotoxicity. Micronucleated peripheral blood cells were counted to assess genotoxicity, and the p21 mRNA level in bone marrow cells was used as a determinant of the p53 regulated DNA damage response. Male mEH-/- mice did not have any significant hematotoxicity or myelotoxicity at the highest benzene exposure compared to the male 129/Sv mice. Significant hematotoxicity or myelotoxicity did not occur in the female mEH-/- or 129/Sv mice. Male mEH-/- mice were also unresponsive to benzene-induced genotoxicity compared to a significant induction in the male 129/Sv mice. The female mEH-/- and 129/Sv mice were virtually unresponsive to benzene-induced genotoxicity. While p21 mRNA expression was highly induced in male 129/Sv mice after exposure to 100-ppm benzene, no significant alteration was observed in male mEH-/- mice. Likewise, p21 mRNA expression in female mEH-/- mice was not significantly induced upon benzene exposure whereas a significant induction was observed in female 129/Sv mice. Thus mEH appears to be critical in benzene-induced toxicity in male, but not female, mice. PMID- 12655034 TI - Thirteen-week inhalation toxicity of N,N-dimethylformamide in F344/N rats and B6C3F1 mice. AB - Male and female F-344 rats and B6C3F1 mice (10/sex/group) were exposed to N,N dimethylformamide (DMF) by whole body inhalation exposure at 0, 50, 100, 200, 400, or 800 ppm, 6 h/day, 5 days/week, for 13 weeks. A concentration-dependent depression in body weight occurred in rats of both sexes at 400 (6-11%) and 800 ppm (20-22%). In contrast, all weight changes in both sexes of mice were within 10% of controls. No rats died, while 5 mice died from nonexposure-related causes. Relative liver weights were significantly increased at all DMF concentrations in both sexes and both species. Activities of serum sorbitol dehydrogenase (SDH) were statistically increased in male and female rats (200 to 800 ppm) on study days 4, 24, and 91 (13 weeks). Activities of alanine aminotransferase (ALT) and isocitrate dehydrogenase (ICD) were statistically increased in both sexes of rats exposed to 800 ppm DMF at all time points. Cholesterol (CHOL) levels were statistically increased in male and female rats (50-800 ppm) at all sampling time points. Levels of total bile acids (TBA) were statistically increased in both sexes of rats (400-800 ppm) on days 24 and 91. Centrilobular hepatocellular necrosis (minimal to moderate) was seen in rats of both sexes exposed at 400 and 800 ppm, with the lesions more severe in females. Centrilobular hepatocellular hypertrophy (minimal to mild) was found in all groups of DMF-exposed male mice, and in female mice exposed at 100-800 ppm. For male and female rats the no observed-adverse-effect concentration (NOAEC) for microscopic liver injury was 200 ppm. The NOAEC was 50 ppm for female mice, but an NOAEC based upon the absence of microscopic liver injury was not determined in male mice. PMID- 12655035 TI - In vitro rat hepatic and intestinal metabolism of the organophosphate pesticides chlorpyrifos and diazinon. AB - Chlorpyrifos (CPF) and diazinon (DZN) are thionophosphorus organophosphate (OP) insecticides; their toxicity is mediated through CYP metabolism to CPF-oxon and DZN-oxon, respectively. Conversely, CYPs also detoxify these OPs to trichloropyridinol (TCP) and 2-isopropyl-4-methyl-6-hydroxypyrimidine (IMHP), respectively. In addition, A-esterase (PON1) metabolism of CPF- and DZN-oxon also forms TCP and IMHP. This study evaluated the role intestinal and hepatic metabolism may play in both the activation and detoxification of CPF and DZN in Sprague-Dawley rats. Similar CYP- and PON1-mediated metabolic profiles were demonstrated in microsomes from liver or isolated intestinal enterocytes. The metabolic efficiency was estimated by calculating the psuedo-1st order rate constant from the metabolic constants by dividing Vmax/Km. In enterocyte microsomes, the CYP metabolic efficiency for metabolism to the oxon metabolites was approximately 28-fold greater for CPF than DZN. Compared on a per nmol P450 basis, the Vmax for CPF in enterocytes was approximately 2-3 times higher than in liver microsomes for the production of CPF-oxon and TCP. The Michaelis-Menten rate constant (Km) for the metabolism of CPF to CPF-oxon was comparable in liver and enterocyte microsomes; however, the enterocyte Km for TCP production was higher (indicating a lower affinity). The smaller volume of intestine, lower amount of CYP, and higher Km for TCP in the enterocyte microsomes, resulted in a lower catalytic efficiency (2 and 62 times) than in liver for oxon and TCP. PON1 mediated metabolism of CPF- and DZN-oxon was also demonstrated in liver and enterocyte microsomes. Although PON1 affinity for the substrates was comparable in hepatic and enterocytic microsomes, the Vmax were 48- to 275-fold higher, in the liver. These results suggest that intestinal metabolism may impact the metabolism of CPF and DZN, especially following low-dose oral exposures. PMID- 12655036 TI - Effects of dibutyl phthalate in male rabbits following in utero, adolescent, or postpubertal exposure. AB - We evaluated sequelae in male rabbits following exposure to dibutyl phthalate (DBP) at a dose level known to adversely affect testicular function in rodents without causing systemic toxicity. Because rabbits have a relatively long phase of reproductive development simulating better than rodents the reproductive development of humans, and because the use of rabbits facilitates multiple evaluations of mating ability and seminal quality, we used this animal model. Rabbits were exposed to 0 or 400 mg DBP/kg/day in utero (gestation days [GD] 15 29) or during adolescence (postnatal weeks [PNW] 4-12), and male offspring were examined at 6, 12, and 25 weeks of age. Another group was exposed after puberty (for 12 weeks) and examined at the conclusion of exposure. The most pronounced reproductive effects were in male rabbits exposed in utero. Male offspring in this group exhibited reduction in numbers of ejaculated sperm (down 43%; p < 0.01), in weights of testes (at 12 weeks, down 23%; p < 0.05) and in accessory sex glands (at 12 and 25 weeks, down 36%; p < 0.01 and down 27%; p < 0.05, respectively). Serum testosterone levels were down (at 6 weeks, 32%; p < 0.05); a slight increase in histological alterations of the testis (p < 0.05) and a doubling in the percentage (from 16 to 30%, p < 0.01) of abnormal sperm; and 1/17 males manifesting hypospadias, hypoplastic prostate, and cryptorchid testes with carcinoma in situ-like cells. In the DBP group exposed during adolescence, basal serum testosterone levels were reduced at 6 weeks (p < 0.01) while at 12 weeks, testosterone production in vivo failed to respond normally to a GnRH challenge (p < 0.01). In addition, weight of accessory sex glands was reduced at 12 weeks but not at 25 weeks after a recovery period; there was a slight increase in the percentage of abnormal sperm in the ejaculate; and 1/11 males was unilaterally cryptorchid. In both of these DBP-treated groups, daily sperm production, epididymal sperm counts, mating ability, and weights of body and nonreproductive organs were unaffected. Thus, DBP induces lesions in the reproductive system of the rabbit, with the intrauterine period being the most sensitive stage of life. PMID- 12655037 TI - Gene expression profile induced by 17 alpha-ethynyl estradiol in the prepubertal female reproductive system of the rat. AB - The profound effects of 17beta-estradiol on cell growth, differentiation, and general homeostasis of the reproductive and other systems, are mediated mostly by regulation of temporal and cell type-specific expression of different genes. In order to understand better the molecular events associated with the activation of the estrogen receptor (ER), we have used microarray technology to determine the transcriptional program and dose-response characteristics of exposure to a potent synthetic estrogen, 17 alpha-ethynyl estradiol (EE), during prepubertal development. Changes in patterns of gene expression were determined in the immature uterus and ovaries of Sprague-Dawley rats on postnatal day (PND) 24, 24 h after exposure to EE, at 0.001, 0.01, 0.1, 1 and 10 micro g EE/kg/day (sc), for four days (dosing from PND 20 to 23). The transcript profiles were compared between treatment groups and controls using oligonucleotide arrays to determine the expression level of approximately 7000 annotated rat genes and over 1740 expressed sequence tags (ESTs). Quantification of the number of genes whose expression was modified by the treatment, for each of the various doses of EE tested, showed clear evidence of a dose-dependent treatment effect that follows a monotonic response, concordant with the dose-response pattern of uterine wet weight gain and luminal epithelial cell height. The number of genes whose expression is affected by EE exposure increases according to dose. At the highest dose tested of EE, we determined that the expression level of over 300 genes was modified significantly (p < or = 0.0001). A dose-dependent analysis of the transcript profile revealed a set of 88 genes whose expression is significantly and reproducibly modified (increased or decreased) by EE exposure (p < or = 0.0001). The results of this study demonstrate that, exposure to a potent estrogenic chemical during prepubertal maturation changes the gene expression profile of estrogen-sensitive tissues. Furthermore, the products of the EE regulated genes identified in these tissues have a physiological role in different intracellular pathways, information that will be valuable to determine the mechanism of action of estrogens. Moreover, those genes could be used as biomarkers to identify chemicals with estrogenic activity. PMID- 12655038 TI - The mutagenic potential of the furylethylene derivative 2-furyl-1-nitroethene in the mouse bone marrow micronucleus test. AB - The compound 2-furyl-1-nitroethene (G-0) has been tested to determine its ability to induce clastogenic or aneugenic effects in vivo, through the induction of micronucleated polychromatic erythrocytes (MNPCE) in mouse bone marrow. Groups of five CD-1 male mice were administered once intraperitoneally at a dose range of 5 20 mg/kg and bone marrow was sampled at 24 and 48 h after the treatment. G-0 was dissolved in corn oil, thus a vehicle control group received only corn oil at 10 ml/kg. The positive control group was administered with cyclophosphamide (40 mg/kg). All animals dosed with the highest concentration of the test agent (20 mg/kg) showed evident clinical symptoms of toxicity. Although evidences of bone marrow toxicity were observed, no statistically significant increases in the incidence of MNPCE over the vehicle control group were observed at any sampling time with any of the assayed doses of the G-0 compound. Cyclophosphamide treatment increased the incidence of MNPCE in all treated animals, demonstrating the sensitivity of the assay conditions in which it was carried out. From the results obtained, it is concluded that the test agent G-0 is neither clastogenic nor aneugenic in the erythrocytes from the bone marrow of treated mice at the doses tested. PMID- 12655039 TI - Comparative metabolism of chrysene and 5-methylchrysene by rat and rainbow trout liver microsomes. AB - We have investigated the metabolism of chrysene (CHR) and 5-methychyrsene (5 MeCHR) by Shasta rainbow trout (Oncorhyncus mykiss) and Long Evans rat liver microsomes to assess the effect of a non-benzo ring methyl substituent on the reactions involved in the metabolism of polycyclic aromatic hydrocarbons (PAHs). Trout as well as rat liver microsomes metabolized both CHR and 5-MeCHR at essentially similar rates, indicating that the methyl substituent does not alter the substrate specificity of the cytochrome P450(s) involved in the metabolism of the two PAHs. Dihydrodiols were the major CHR metabolites formed by both trout and rat liver microsomes, whereas the trout liver microsomes formed a considerably higher proportion of 5-MeCHR phenols compared to diols, indicating that 5-methyl substitution alters the substrate specificity of trout microsomal epoxide hydrolase for 5-MeCHR epoxides. Unlike trout liver microsomes, rat liver microsomes formed a much greater proportion of 5-MeCHR diols compared to 5-MeCHR phenols, suggesting that 5-MeCHR epoxides are better substrates for the microsomal epoxide hydrolase present in rat liver than for the enzyme in trout liver. Both trout and rat liver microsomes are more efficient at attacking the bay-region bond versus the non-bay-region double bond in chrysene. In contrast the reverse is true in the case of 5-MeCHR, indicating that a non-benzo ring methyl substituent alters the regioselectivity of the enzymes involved in the oxidative metabolism of PAHs. PMID- 12655040 TI - The legacy of Leon Golberg (1915-1987). PMID- 12655041 TI - The structural basis for biphasic kinetics in the folding of the WW domain from a formin-binding protein: lessons for protein design? AB - The mechanism of formation of beta-sheets is of great importance because of the significant role of such structures in the initiation and propagation of amyloid diseases. In this study we examine the folding of a series of three-stranded antiparallel beta-sheets known as WW domains. Whereas other WW domains have been shown to fold with single-exponential kinetics, the WW domain from murine formin binding protein 28 has recently been shown to fold with biphasic kinetics. By using a combination of kinetics and thermodynamics to characterize a simple model for this protein, the origins of the biphasic kinetics is found to lie in the fact that most of the protein is able to fold without requiring one of the beta hairpins to be correctly registered. The correct register of this hairpin is enforced by a surface-exposed hydrophobic contact, which is not present in other WW domains. This finding suggests the use of judiciously chosen surface-exposed hydrophobic pairs as a protein design strategy for enforcing the desired strand registry. PMID- 12655042 TI - The history of effective population size and genetic diversity in the Yellowstone grizzly (Ursus arctos): implications for conservation. AB - Protein, mtDNA, and nuclear microsatellite DNA analyses have demonstrated that the Yellowstone grizzly bear has low levels of genetic variability compared with other Ursus arctos populations. Researchers have attributed this difference to inbreeding during a century of anthropogenic isolation and population size reduction. We test this hypothesis and assess the seriousness of genetic threats by generating microsatellite data for 110 museum specimens collected between 1912 and 1981. A loss of variability is detected, but it is much less severe than hypothesized. Variance in allele frequencies over time is used to estimate an effective population size of approximately 80 across the 20th century and >100 currently. The viability of the population is unlikely to be substantially reduced by genetic factors in the next several generations. However, gene flow from outside populations will be beneficial in avoiding inbreeding and the erosion of genetic diversity in the future. PMID- 12655043 TI - Translational control of inducible nitric oxide synthase expression by arginine can explain the arginine paradox. AB - L-Arginine is the only endogenous nitrogen-containing substrate of NO synthase (NOS), and it thus governs the production of NO during nervous system development as well as in disease states such as stroke, multiple sclerosis, Parkinson's disease, and HIV dementia. The "arginine paradox" refers to the dependence of cellular NO production on exogenous L-arginine concentration despite the theoretical saturation of NOS enzymes with intracellular L-arginine. Herein, we report that decreased availability of L-arginine blocked induction of NO production in cytokine-stimulated astrocytes, owing to inhibition of inducible NOS (iNOS) protein expression. However, activity of the promoter of the iNOS gene, induction of iNOS mRNA, and stability of iNOS protein were not inhibited under these conditions. Our results indicate that inhibition of iNOS activity by arginine depletion in stimulated astrocyte cultures occurs via inhibition of translation of iNOS mRNA. After stimulation by cytokines, uptake of L-arginine negatively regulates the phosphorylation status of the eukaryotic initiation factor (eIF2 alpha), which, in turn, regulates translation of iNOS mRNA. eIF2 alpha phosphorylation correlates with phosphorylation of the mammalian homolog of yeast GCN2 eIF2 alpha kinase. As the kinase activity of GCN2 is activated by phosphorylation, these findings suggest that GCN2 activity represents a proximal step in the iNOS translational regulation by availability of l-arginine. These results provide an explanation for the arginine paradox for iNOS and define a distinct mechanism by which a substrate can regulate the activity of its associated enzyme. PMID- 12655044 TI - Visualizing the Ca2+-dependent activation of gelsolin by using synchrotron footprinting. AB - Radiolytic protein footprinting with a synchrotron source is used to reveal detailed structural changes that occur in the Ca(2+)-dependent activation of gelsolin. More than 80 discrete peptides segments within the structure, covering 95% of the sequence in the molecule, were examined by footprinting and mass spectrometry for their solvent accessibility as a function of Ca(2+) concentration in solution. Twenty-two of the peptides exhibited detectable oxidation; for seven the oxidation extent was seen to be Ca(2+) sensitive. Ca(2+)titration isotherms monitoring the oxidation within residues 49-72 (within subdomain S1), 121-135 (S1), 162-166 (S2), and 722-748 (S6) indicate a three state activation process with a intermediate that was populated at a Ca(2+) concentration of 1-5 microM that is competent for capping and severing activity. A second structural transition with a midpoint of approximately 60-100 microM, where the accessibility of the above four peptides is further increased, is also observed. Tandem mass spectrometry showed that buried residues within the helical "latch" of S6 (including Pro-745) that contact an F-actin-binding site on S2 and buried F-actin-binding residues within S2 (including Phe-163) are unmasked in the submicromolar Ca(2+) transition. However, residues within S4 that are part of an extended beta-sheet with S6 (including Tyr-453) are revealed only in the subsequent transition at higher Ca(2+) concentrations; the disruption of this extended contact between S4 and S6 (and likely the analogous contact between S1 and S3) likely results in an extended structure permitting additional functions consistent with the fully activated gelsolin molecule. PMID- 12655045 TI - Macula densa cell signaling involves ATP release through a maxi anion channel. AB - Macula densa cells are unique renal biosensor cells that detect changes in luminal NaCl concentration ([NaCl](L)) and transmit signals to the mesangial cellafferent arteriolar complex. They are the critical link between renal salt and water excretion and glomerular hemodynamics, thus playing a key role in regulation of body fluid volume. Since identification of these cells in the early 1900s, the nature of the signaling process from macula densa cells to the glomerular contractile elements has remained unknown. In patch-clamp studies of macula densa cells, we identified an [NaCl](L)-sensitive ATP-permeable large conductance (380 pS) anion channel. Also, we directly demonstrated the release of ATP (up to 10 microM) at the basolateral membrane of macula densa cells, in a manner dependent on [NaCl](L), by using an ATP bioassay technique. Furthermore, we found that glomerular mesangial cells respond with elevations in cytosolic Ca(2+) concentration to extracellular application of ATP (EC(50) 0.8 microM). Importantly, we also found increases in cytosolic Ca(2+) concentration with elevations in [NaCl](L), when fura-2-loaded mesangial cells were placed close to the basolateral membrane of macula densa cells. Thus, cell-to-cell communication between macula densa cells and mesangial cells, which express P2Y(2) receptors, involves the release of ATP from macula densa cells via maxi anion channels at the basolateral membrane. This mechanism may represent a new paradigm in cell-to cell signal transduction mediated by ATP. PMID- 12655047 TI - Toll-like receptor 4 on stromal and hematopoietic cells mediates innate resistance to uropathogenic Escherichia coli. AB - Innate host defenses at mucosal surfaces are critical in the early stages of many bacterial infections. In addition to cells of the traditional innate immune system, epithelial cells can also produce inflammatory mediators during an infection. However, the role of the epithelium in innate host defense in vivo is unclear. Recent studies have shown that lipopolysaccharide (LPS) recognition is critical for bladder epithelial cells to recognize and respond to Escherichia coli. Moreover, the LPS-nonresponsive mouse strain C3HHeJ, which has a mutation in the primary LPS receptor, Toll-like receptor 4 (TLR4), is extremely susceptible to infection with uropathogenic strains of E. coli. In this study, a bone marrow transplant approach was used to investigate the specific contributions of the bladder epithelium (and other stromal cells) in the TLR4 mediated innate immune response to the invading E. coli pathogen. Mice expressing the mutant TLR4 in the epithelialstromal compartment were not able to mount a protective inflammatory response to control the early infection even when their hematopoietic cells expressed wild-type TLR4. However, the presence of TLR4(+) epithelialstromal cells was not sufficient to activate an acute inflammatory response unless the hematopoietic cells were also TLR4(+). These results demonstrated that bladder epithelial cells play a critical role in TLR4-mediated innate immunity in vivo during a mucosal bacterial infection. PMID- 12655046 TI - A metal-binding site in the catalytic subunit of anaerobic ribonucleotide reductase. AB - A Zn(Cys)(4) center has been found in the C-terminal region of the crystal structure of the anaerobic class III ribonucleotide reductase (RNR) from bacteriophage T4. The metal center is structurally related to the zinc ribbon motif and to rubredoxin and rubrerythrin. Mutant enzymes of the homologous RNR from Escherichia coli, in which the coordinating cysteines, conserved in almost all known class III RNR sequences, have been mutated into alanines, are shown to be inactive as the result of their inability to generate the catalytically essential glycyl radical. The possible roles of the metal center are discussed in relationship to the currently proposed reaction mechanism for generation of the glycyl radical in class III RNRs. PMID- 12655048 TI - Bioinorganic chemistry in the postgenomic era. AB - Genome sequencing has revolutionized all fields of life sciences. Bioinorganic chemistry is certainly not immune to this influence, which is presenting unprecedented challenges. A new goal for bioinorganic chemistry is the investigation of the linkages between inorganic elements and genomic information. This requires new advancements andor the development of new expertise in fields such as bioinformatics and genetics but also provides a driving force to push forward the exploitation of traditional analytical techniques and spectroscopic tools. The "case study" of metal homeostasis in cells is discussed to provide a flavor of the current evolution of the field. PMID- 12655049 TI - Neutral thiol as a proximal ligand to ferrous heme iron: implications for heme proteins that lose cysteine thiolate ligation on reduction. AB - Cysteine plays a key role as a metal ligand in metalloproteins. In all well recognized cases, however, it is the anionic cysteinate that coordinates. Several cysteinate-ligated heme proteins are known, but some fail to retain thiolate ligation in the ferrous state, possibly following protonation to form neutral cysteine. Ligation by cysteine thiol in ferrous heme proteins has not been documented. To establish spectroscopic signatures for such systems, we have prepared five-coordinate adducts of the ferrous myoglobin H94G cavity mutant with neutral thiol and thioether sulfur donors as well as six-coordinate derivatives such as with CO and, when possible, with NO and O(2). A thiol-ligated oxyferrous complex is reported, to our knowledge for the first time. Further, a bis thioether ferrous H93G model for bis-methionine ligation, as found in Pseudomonas aeruginosa bacterioferritin heme protein, is described. Magnetic CD spectroscopy has been used due to its established ability in axial ligand identification. The magnetic CD spectra of the H93G complexes have been compared with those of ferrous H175CD235L cytochrome c peroxidase to show that its proximal ligand is neutral cysteine. We had previously reported this cytochrome c peroxidase mutant to be cysteinate-ligated in the ferric state, but the ferrous ligand was undetermined. The spectral properties of ferrous liver microsomal cytochrome P420 (inactive P450) are also consistent with thiol ligation. This study establishes that neutral cysteine can serve as a ligand in ferrous heme iron proteins, and that ferric cysteinate-ligated heme proteins that fail to retain such ligation on reduction may simply be ligated by neutral cysteine. PMID- 12655050 TI - Superoxo, mu-peroxo, and mu-oxo complexes from heme/O2 and heme-Cu/O2 reactivity: copper ligand influences in cytochrome c oxidase models. AB - The O(2)-reaction chemistry of 1:1 mixtures of (F(8))Fe(II) (1; F(8) = tetrakis(2,6-diflurorophenyl)porphyrinate) and [(L(Me(2))N)Cu(I)](+) (2; L(Me(2))N = N,N-bis(2-[2-(N',N'-4-dimethylamino)pyridyl]ethyl)methylamine) is described, to model aspects of the chemistry occurring in cytochrome c oxidase. Spectroscopic investigations, along with stopped-flow kinetics, reveal that low temperature oxygenation of 1/2 leads to rapid formation of a heme-superoxo species (F(8))Fe(III)-(O(2)(-)) (3), whether or not 2 is present. Complex 3 subsequently reacts with 2 to form [(F(8))Fe(III)-(O(2)(2-)) Cu(II)(L(Me(2))N)](+) (4), which thermally converts to [(F(8))Fe(III)-(O) Cu(II)(L(Me(2))N)](+) (5), which has an unusually bent (Fe-O-Cu) bond moiety. Tridentate chelation, compared with tetradentate, is shown to dramatically lower the nu(O-O) values observed in 4 and give rise to the novel structural features in 5. PMID- 12655051 TI - New clues for platinum antitumor chemistry: kinetically controlled metal binding to DNA. AB - From the metal ions and metal compounds that are known to bind to DNA, many anticancer Pt(II) and Ru(II)Ru(III) compounds are known to have ligand-exchange kinetics in the same order of magnitude as the division of tumor cells. The present article discusses this process in detail with special attention to cisplatin and related compounds and the cellular binding sites and processes of such compounds. Detailed platinated DNA structures are presented and discussed in light of the mechanistic studies of metal antitumor compounds. It is now known that platinum antitumor drugs eventually end up on the DNA. However, it remains a challenge to understand how (fast) they reach the DNA and how they are removed. The kinetics of ligand exchange around platinum appear to play a crucial role, and the possible role of other ligands as intermediates, especially those with S donor sites, is of great interest. New types of Pt compounds with additional functionalities influencing DNA binding and kinetics are discussed in the context of steric and H-bonding properties. A comparison is made with more sterically crowded Ru complexes. The effects on activity and correlations with structural and kinetic properties are clues in understanding the biological activities of these classes of compounds. PMID- 12655053 TI - Salmochelins, siderophores of Salmonella enterica and uropathogenic Escherichia coli strains, are recognized by the outer membrane receptor IroN. AB - Members of a family of catecholate siderophores, called salmochelins, were isolated by reversed-phase HPLC from Salmonella enterica serotype Typhimurium and structurally characterized by Fourier transform ion cyclotron resonance-MSMS and GC-MS. The tentative structure of salmochelin 1 contained two 2,3- dihydroxybenzoylserine moieties bridged by a glucose residue, bound to the serine hydroxyl group of one moiety and the carboxylate of the second moiety. Salmochelin 2 contained in addition a second glucose residue linked to a third 2,3-dihydroxybenzoylserine moiety. Salmochelins were not produced by an iroBC mutant, which indicated that the IroB protein might be responsible for the glucosyl transfer predicted by sequence similarities to known glycosyltransferases. Uptake experiments with radiolabeled (55)Fe-salmochelin and growth promotion tests with salmochelins showed that the IroN outer membrane receptor, encoded in the iroA locus of S. enterica and uropathogenic Escherichia coli strains, was the main receptor for ferric salmochelin transport. PMID- 12655052 TI - The role of copper and protons in heme-copper oxidases: kinetic study of an engineered heme-copper center in myoglobin. AB - To probe the role of copper and protons in heme-copper oxidase (HCO), we have performed kinetic studies on an engineered heme-copper center in sperm whale myoglobin (Leu-29 --> HisPhe-43 --> His, called Cu(B)Mb) that closely mimics the heme-copper center in HCO. In the absence of metal ions, the engineered Cu(B) center in Cu(B)Mb decreases the O(2) binding affinity of the heme. However, addition of Ag(I), a redox-inactive mimic of Cu(I), increases the O(2)-binding affinity. More importantly, copper ion in the Cu(B) center is essential for O(2) reduction, as no O(2) reduction can be observed in copper-free, Zn(II), or Ag(I) derivatives of Cu(B)Mb. Instead of producing a ferryl-heme as in HCO, the Cu(B)Mb generates verdoheme because the engineered Cu(B)Mb may lack a hydrogen bonding network that delivers protons to promote the heterolytic OO cleavage necessary for the formation of ferryl-heme. Reaction of oxidized Cu(B)Mb with H(2)O(2), a species equivalent in oxidation state to 2e(-), reduced O(2) but, possessing the extra protons, resulted in ferryl-heme formation, as in HCO. The results showed that the Cu(B) center plays a critical role in O(2) binding and reduction, and that proton delivery during the O(2) reduction is important to avoid heme degradation and to promote the HCO reaction. PMID- 12655054 TI - Metal stoichiometry and functional studies of the diphtheria toxin repressor. AB - Diphtheria toxin repressor (DtxR) is a transition metal ion-activated repressor in Corynebacterium diphtheriae. DtxR is an iron sensor; metal-bound DtxR represses transcription of genes downstream of the tox operator. Wild-type DtxR [DtxR(wt)] and several mutant forms were overexpressed and purified from Escherichia coli. DtxR was isolated without bound metal. Metal reconstitution gave a binding stoichiometry of 2 per monomer for DtxR(wt) and 1 per monomer for DtxR(H79A) and DtxR(M10A). DNA binding of DtxR(H79A) and DtxR(M10A) indicates that metal site 2 is essential for activity. Metal binding lowers the dimerization K(d) of DtxR from low micromolar to 33 nM. Gel electrophoretic mobility-shift assays show that Fe(2+) and not Fe(3+) activates DtxR for DNA binding. This finding suggests that gene regulation by DtxR may be sensitive not only to iron levels but also to redox state of the iron. Mutations in the tox operator sequence indicate that DtxR dimers binding to DNA may be highly cooperative. PMID- 12655055 TI - The netrin-1 receptors UNC5H are putative tumor suppressors controlling cell death commitment. AB - The three mammalian receptors UNC5H1, UNC5H2, and UNC5H3 (also named UNC5A, UNC5B, and UNC5C in human) that belong to the family of the netrin-1 receptors, UNC5H, were initially proposed as mediators of the chemorepulsive effect of netrin-1 on specific axons. However, they were also recently shown to act as dependence receptors. Such receptors induce apoptosis when unbound to their ligand. We show here that the expression of the human UNC5A, UNC5B, or UNC5C is down-regulated in multiple cancers including colorectal, breast, ovary, uterus, stomach, lung, or kidney cancers. In colorectal tumors, this down-regulation is associated with loss of heterozygosity occurring within UNC5A, UNC5B, and UNC5C genes but may also be partially related to epigenetic processes because histone deacetylase inhibitor increased UNC5C expression in various cancer cell lines. Moreover, sequencing of UNC5C gene in patients with colorectal tumors revealed the presence of missense mutations. The lossreduction of expression may be a crucial mechanism for tumorigenicity because the expression of UNC5H1, UNC5H2, or UNC5H3 inhibits tumor cell anchorage-independent growth and invasion. Moreover, these hallmarks of malignant transformation can be restored by netrin-1 addition or apoptosis inhibition. Hence, UNC5H1, UNC5H2, and UNC5H3 receptors may represent tumor suppressors that inhibit tumor extension outside the region of netrin-1 availability by inducing apoptosis. PMID- 12655056 TI - The bioinorganic chemistry of iron in oxygenases and supramolecular assemblies. AB - The bioinorganic chemistry of iron is central to life processes. Organisms must recruit iron from their environment, control iron storage and trafficking within cells, assemble the complex, iron-containing redox cofactors of metalloproteins, and manage a myriad of biochemical transformations by those enzymes. The coordination chemistry and the variable oxidation states of iron provide the essential mechanistic machinery of this metabolism. Our current understanding of several aspects of the chemistry of iron in biology are discussed with an emphasis on the oxygen activation and transfer reactions mediated by heme and nonheme iron proteins and the interactions of amphiphilic iron siderophores with lipid membranes. PMID- 12655057 TI - Evidence against the presence of an anandamide transporter. AB - On the basis of temperature dependency, saturability, selective inhibition, and substrate specificity, it has been proposed that an anandamide transporter exists. However, all of these studies have examined anandamide accumulation at long time points when downstream effects such as metabolism and intracellular sequestration are operative. In the current study, we have investigated the initial rates (<1 min) of anandamide accumulation in neuroblastoma and astrocytoma cells in culture and have determined that uptake is not saturable with increasing concentrations of anandamide. However, anandamide hydrolysis, after uptake in neuroblastoma cells, was saturable at steady-state time points (5 min), suggesting that fatty acid amide hydrolase (FAAH) may be responsible for observed saturation of uptake at long time points. In general, arvanil, olvanil, and N-(4-hydroxyphenyl)arachidonylamide (AM404) have been characterized as transport inhibitors in studies using long incubations. However, we found these "transport inhibitors" did not inhibit anandamide uptake in neuroblastoma and astrocytoma cells at short time points (40 sec or less). Furthermore, we confirmed that these inhibitors in vitro were actually inhibitors of FAAH. Therefore, the likely mechanism by which the transport inhibitors raise anandamide levels to exert pharmacological effects is by inhibiting FAAH, and they should be reevaluated in this context. Immunofluorescence has indicated that FAAH staining resides mainly on intracellular membranes of neuroblastoma cells, and this finding is consistent with our observed kinetics of anandamide hydrolysis. In summary, these data suggest that anandamide uptake is a process of simple diffusion. This process is driven by metabolism and other downstream events, rather than by a specific membrane-associated anandamide carrier. PMID- 12655058 TI - Pitx3 is required for development of substantia nigra dopaminergic neurons. AB - Dopaminergic (DA) neurons of substantia nigra in the midbrain control voluntary movement, and their degeneration is the cause of Parkinson's disease. The complete set of genes required to specifically determine the development of midbrain DA subgroups is not known yet. We report here that mice lacking the bicoid-related homeoprotein Pitx3 fail to develop DA neurons of the substantia nigra. Other mesencephalic DA neurons of the ventral tegmental area and retrorubral field are unaltered in their dopamine expression and histological organization. These data suggest that Pitx3-dependent gene expression is specifically required for the differentiation of DA progenitors within the mesencephalic DA system. PMID- 12655059 TI - Mouse ribonucleotide reductase R2 protein: a new target for anaphase-promoting complex-Cdh1-mediated proteolysis. AB - Ribonucleotide reductase consists of two nonidentical proteins, R1 and R2, and catalyzes the rate-limiting step in DNA precursor synthesis: the reduction of ribonucleotides to deoxyribonucleotides. A strictly balanced supply of deoxyribonucleotides is essential for both accurate DNA replication and repair. Therefore, ribonucleotide reductase activity is under exquisite control both transcriptionally and posttranscriptionally. In proliferating mammalian cells, enzyme activity is regulated by control of R2 protein stability. This control, which responds to DNA damage, is effective until cells pass into mitosis. We demonstrate that the mitotic degradation and hence the overall periodicity of R2 protein levels depends on a KEN box sequence, recognized by the Cdh1-anaphase promoting complex. The mouse R2 protein specifically binds Cdh1 and is polyubiquitinated in an in vitro ubiquitin assay system. Mutating the KEN signal stabilizes the R2 protein during mitosisG(1) in R2 protein-overexpressing cells. The degradation process, which blocks deoxyribonucleotide production during G(1), may be an important mechanism protecting the cell against unscheduled DNA synthesis. The newly discovered p53-induced p53R2 protein that lacks a KEN box may supply deoxyribonucleotides for DNA repair during G(0)G(1). PMID- 12655060 TI - Critical role of oxidative stress in estrogen-induced carcinogenesis. AB - Mechanisms of estrogen-induced tumorigenesis in the target organ are not well understood. It has been suggested that oxidative stress resulting from metabolic activation of carcinogenic estrogens plays a critical role in estrogen-induced carcinogenesis. We tested this hypothesis by using an estrogen-induced hamster renal tumor model, a well established animal model of hormonal carcinogenesis. Hamsters were implanted with 17beta-estradiol (betaE2), 17alpha-estradiol (alphaE2), 17alpha-ethinylestradiol (alphaEE), menadione, a combination of alphaE2 and alphaEE, or a combination of alphaEE and menadione for 7 months. The group treated with betaE2 developed target organ specific kidney tumors. The kidneys of hamsters treated with alphaE2, alphaEE, or menadione alone did not show any gross evidence of tumor. Kidneys of hamsters treated with a combination of alphaE2 and alphaEE showed early signs of proliferation in the interstitial cells. Kidneys of hamsters treated with a combination of menadione and alphaEE showed foci of tumor with congested tubules and atrophic glomeruli. betaE2 treated tumor-bearing kidneys showed >2-fold increase in 8-iso-prostaglandin F(2alpha) (8-iso-PGF(2alpha)) levels compared with untreated controls. Kidneys of hamsters treated with a combination of menadione and alphaEE showed increased 8 iso-PGF(2alpha) levels compared with untreated controls, whereas no increase in 8 iso-PGF(2alpha) was detected in kidneys of alphaEE-treated group. A chemical known to produce oxidative stress or a potent estrogen with poor ability to produce oxidative stress, were nontumorigenic in hamsters, when given as single agents, but induced renal tumors, when given together. Thus, these data provide evidence that oxidant stress plays a crucial role in estrogen-induced carcinogenesis. PMID- 12655061 TI - Recombination signal sequence-binding protein Jkappa alters mesodermal cell fate decisions by suppressing cardiomyogenesis. AB - The transcription factor recombination signal sequence-binding protein Jkappa (RBP-J) is a key downstream element in the signaling pathway of all four mammalian Notch receptors that are critically involved in the control of embryonic and adult development. RBP-J-deficient mice display complex defects and die around day 9.5 postcoitum. Here, we investigate the function of RBP-J in the development of mesodermal cell lineages by using the OP9 stroma coculture system. RBP-J-deficient embryonic stem (ES) cells gave rise to cardiomyocytes, endothelial cells, and primitive and definitive hematopoietic cells. Thus, RBP-J mediated signals are not required for generation of these cell types. However, when compared with parental RBP-J-expressing ES cells, cardiomyogenesis derived from RBP-J-deficient ES cells was increased. Repression over the cardiogenic pathway was restored by expressing RBP-J in RBP-J-deficient ES cells. Our data indicate that Notch signaling via RBP-J plays an important role for the correct specification of myocardial cell fates. PMID- 12655062 TI - Enterobactin: an archetype for microbial iron transport. AB - Bacteria have aggressive acquisition processes for iron, an essential nutrient. Siderophores are small iron chelators that facilitate cellular iron transport. The siderophore enterobactin is a triscatechol derivative of a cyclic triserine lactone. Studies of the chemistry, regulation, synthesis, recognition, and transport of enterobactin make it perhaps the best understood of the siderophore mediated iron uptake systems, displaying a lot of function packed into this small molecule. However, recent surprises include the isolation of corynebactin, a closely related trithreonine triscatechol derivative lactone first found in Gram positive bacteria, and the crystal structure of a ferric enterobactin complex of a protein identified as an antibacterial component of the human innate immune system. PMID- 12655064 TI - Identification and functional characterization of a novel binding site on TNF alpha promoter. AB - Transcription of the tumor necrosis factor (TNF) gene is rapidly and transiently induced by lipopolysaccharide in cells of monocytemacrophage lineage. Previous studies have suggested that in the mouse, multiple NF-kappaBRel-binding sites contribute to the TNF transcriptional response to LPS. But the role of these regulatory elements in transcriptional activation of the TNF-alpha gene in human monocytes remains unclear. Previously, a transcription factor, termed lipopolysaccharide-induced TNF-alpha factor (LITAF), was found to regulate TNF alpha gene expression. However, the specific protein domain(s) of human (h)LITAF that interact with the hTNF-alpha promoter had not been identified. In this study, we identify by footprinting a sequence motif, CTCCC (-515 to -511), within the TNF-alpha promoter that binds to hLITAF. We also identify the region of hLITAF (amino acids 165-180) that was named peptide B and specifically mediates binding to the hTNF-alpha promoter. When THP-1 cells were stimulated with this peptide B, it was sufficient to induce TNF-alpha secretion. Induction of TNF alpha transcription by LPS or peptide B depended on the presence of the -515 to 511 promoter region, which was found to be essential for hLITAF binding. Together, these findings help to clarify the mechanism of hLITAFhTNF-alpha interaction and the manner by which hLITAF contributes to hTNF-alpha regulation in an attempt to design new pharmacological interventions to address TNF-related diseases. PMID- 12655063 TI - Analysis of the eye developmental pathway in Drosophila using DNA microarrays. AB - Pax-6 genes encode evolutionarily conserved transcription factors capable of activating the gene-expression program required to build an eye. When ectopically expressed in Drosophila imaginal discs, Pax-6 genes induce the eye formation on the corresponding appendages of the adult fly. We used two different Drosophila full-genome DNA microarrays to compare gene expression in wild-type leg discs versus leg discs where eyeless, one of the two Drosophila Pax-6 genes, was ectopically expressed. We validated these data by analyzing the endogenous expression of selected genes in eye discs and identified 371 genes that are expressed in the eye imaginal discs and up-regulated when an eye morphogenetic field is ectopically induced in the leg discs. These genes mainly encode transcription factors involved in photoreceptor specification, signal transducers, cell adhesion molecules, and proteins involved in cell division. As expected, genes already known to act downstream of eyeless during eye development were identified, together with a group of genes that were not yet associated with eye formation. PMID- 12655065 TI - Context sensitivity of activity-dependent increases in cerebral blood flow. AB - Functional neuroimaging in humans is used widely to study brain function in relation to human disease and cognition. The neural basis of neuroimaging signals is probably synaptic activity, but the effect of context, defined as the interaction between synaptic inhibition, excitation, and the electroresponsive properties of the targeted neurons, is not well understood. We examined here the effect of interaction of synaptic excitation and net inhibition on the relationship between electrical activity and vascular signals in the cerebellar cortex. We show that stimulation of the net inhibitory parallel fibers simultaneously with stimulation of the excitatory climbing fibers leads to a further rise in total local field potentials (LFP) and cerebral blood flow (CBF) amplitudes, not a decrease, as predicted from theoretical studies. However, the combined stimulation of the parallel and climbing fiber systems produced changes in CBF and LFP that were smaller than their algebraic sum evoked by separate stimulation of either system. This finding was independent of the starting condition, i.e., whether inhibition was superimposed on a state of excitation or vice versa. The attenuation of the increases in LFP and CBF amplitudes was similar, suggesting that synaptic activity and CBF were coupled under these conditions. The result might be explained by a relative neuronal refractoriness that relates to the intrinsic membrane properties of Purkinje cells, which determine the recovery time of these cells. Our work implies that neuronal and vascular signals are context-sensitive and that their amplitudes are modulated by the electroresponsive properties of the targeted neurons. PMID- 12655066 TI - Investigation of metal-dithiolate fold angle effects: implications for molybdenum and tungsten enzymes. AB - Gas-phase photoelectron spectroscopy and density functional theory have been used to investigate the interactions between the sulfur pi-orbitals of arene dithiolates and high-valent transition metals as minimum molecular models of the active site features of pyranopterin MoW enzymes. The compounds (Tp*)MoO(bdt) (compound 1), Cp(2)Mo(bdt) (compound 2), and Cp(2)Ti(bdt) (compound 3) [where Tp* is hydrotris(3,5-dimethyl-1-pyrazolyl)borate, bdt is 1,2-benzenedithiolate, and Cp is eta(5)- cyclopentadienyl] provide access to three different electronic configurations of the metal, formally d(1), d(2), and d(0), respectively. The gas phase photoelectron spectra show that ionizations from occupied metal and sulfur based valence orbitals are more clearly observed in compounds 2 and 3 than in compound 1. The observed ionization energies and characters compare very well with those calculated by density functional theory. A "dithiolate-folding-effect" involving an interaction of the metal in-plane and sulfur-pi orbitals is proposed to be a factor in the electron transfer reactions that regenerate the active sites of molybdenum and tungsten enzymes. PMID- 12655067 TI - Nitric oxide binding at the mononuclear active site of reduced Pyrococcus furiosus superoxide reductase. AB - Nitric oxide (NO) has been used as a substrate analog to explore the structural and electronic determinants of enzymatic superoxide reduction at the mononuclear iron active site of Pyrococcus furiosus superoxide reductase (SOR) through the use of EPR, resonance Raman, Fourier transform IR, UV-visible absorption, and variable-temperature variable-field magnetic CD spectroscopies. The NO adduct of reduced SOR is shown to have a near-axial S = 32 ground state with ED = 0.06 and D = 12 +/- 2 cm(-1) (where D and E are the axial and rhombic zero-field splitting parameters, respectively) and the UV-visible absorption and magnetic CD spectra are dominated by an out-of-plane NO(-)(pi*)-to-Fe(3+)(dpi) charge-transfer transition, polarized along the zero-field splitting axis. Resonance Raman studies indicate that the NO adduct is six-coordinate with NO ligated in a bent conformation trans to the cysteinyl S, as evidenced by the identification of nu(N O) at 1,721 cm(-1), nu(Fe-NO) at 475 cm(-1), and nu(Fe-S(Cys), at 291 cm(-1), via (34)S and (15)NO isotope shifts. The electronic and vibrational properties of the S = 32 (FeNO)(7) unit are rationalized in terms of a limiting formulation involving a high-spin (S = 52) Fe(3+) center antiferromagnetically coupled to a (S = 1) NO(-) anion, with a highly covalent Fe(3+)-NO(-) interaction. The results support a catalytic mechanism for SOR, with the first step involving oxidative addition of superoxide to form a ferric-peroxo intermediate, and indicate the important roles that the Fe spin state and the trans cysteinate ligand play in effecting superoxide reduction and peroxide release. PMID- 12655068 TI - How does cyanide inhibit superoxide reductase? Insight from synthetic FeIIIN4S model complexes. AB - Superoxide reductases (SORs) are nonheme iron-containing enzymes that reduce HO(2) to H(2)O(2). Exogenous substrates such as N(3)(-) and CN(-) have been shown to bind to the catalytic iron site of SOR, and cyanide acts as an inhibitor. To understand how these exogenous ligands alter the physical and reactivity properties of the SOR iron site, acetate-, azide-, and cyanide-ligated synthetic models of SOR have been prepared. The x-ray crystal structures of azide-ligated [Fe(III)(S(Me2)N(4)(tren))(N(3))](+) (3), dimeric cyanide-bridged ([Fe(III)(S(Me2)N(4)(tren))](2)-mu-CN)(3+) (5), and acetate-ligated [Fe(III)(S(Me2)N(4)(tren))(OAc)](+) (6) are described, in addition to x-ray absorption spectrum-derived and preliminary crystallographic structures of cyanide-ligated [Fe(III)(S(Me2)N(4)(tren))(CN)](+) (4). Cyanide coordination to our model (4) causes the redox potential to shift anodically by 470 mV relative to acetate-ligated 6 and 395 mV relative to azide-ligated 3. If cyanide coordination were to cause a similar shift in redox potential with SOR, then the reduction potential of the catalytically active Fe(3+) center would fall well below that of its biological reductants. These results suggest therefore that cyanide inhibits SOR activity by making the Fe(2+) state inaccessible and thus preventing the enzyme from turning over. Cyanide inhibits activity in the metalloenzyme superoxide dismutase via a similar mechanism. The reduced five coordinate precursor to 3, 4, and 6 [Fe(II)(S(Me2)N(4)(tren))](+) (1) was previously shown by us to react with superoxide to afford H(2)O(2) via an [Fe(III)(S(Me2)N(4)(tren))(OOH)](+) intermediate. Cyanide and azide do not bind to 1 and do not prevent 1 from reducing superoxide. PMID- 12655069 TI - Meeting of the minds: metalloneurochemistry. AB - Metalloneurochemistry is the study of metal ion function in the brain and nervous system at the molecular level. Research in this area is exemplified through discussion of several forefront areas where significant progress has been made in recent years. The structure and function of ion channels have been elucidated through high-resolution x-ray structural work on the bacterial K(+) ion channel. Selection of potassium over sodium ions is achieved by taking advantage of key principles of coordination chemistry. The role of calcium ions in neuronal signal transduction is effected by several Ca(2+)-binding protein such as calmodulin, calcineurin, and synaptotagmin. Structural changes in response to calcium ion concentrations allow these proteins to function in memory formation and other neurochemical roles. Metallochaperones help to achieve metal ion homeostasis and thus prevent neurological diseases because of metal ion imbalance. Much detailed chemical information about these systems has become available recently. Zinc is another important metal ion in neuroscience. Its concentration in brain is in part controlled by metallothionein, and zinc is released in the hippocampus at glutamatergic synapses. New fluorescent sensors have become available to help track such zinc release. PMID- 12655071 TI - Male-biased transmission of deleterious mutations to the progeny in Arabidopsis thaliana. AB - The extent and cause of male-biased mutation rates, the higher number of mutations in sperm than in eggs, is currently an active and controversial subject. Recent evidence indicates that this male (sperm) bias not only occurs in animals but also in plants. The higher mutation rate in plant sperm was inferred from rates of evolution of neutral DNA regions, and the results were confined to the mitochondria and chloroplasts of gymnosperms. However, the relative transmission rates of deleterious mutations, which have substantial evolutionary consequences, have rarely been studied. Here, an investigation is described by using the hermaphroditic self-compatible flowering plant Arabidopsis thaliana, in which we artificially increased the rate of mutation in pollen (i.e., sperm donor) and maternal (i.e., egg donor) parents, by using two kinds of UV irradiation in parallel and separate experiments, and assessed the deleterious effects on fitness of the F(2) generation. The results show that more deleterious induced mutations are transmitted to the progeny by a sperm than by an egg. These findings provide the first experimental evidence that more deleterious mutations are inherited from sperm than from an egg in any organism. Possible causes underlying this male bias are discussed. PMID- 12655070 TI - Misfolded CuZnSOD and amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a progressive degenerative disease of motor neurons. The inherited form of the disease, familial ALS, represents 5-10% of the total cases, and the best documented of these are due to lesions in SOD1, the gene encoding copper-zinc superoxide dismutase (CuZnSOD). The mechanism by which mutations in SOD1 cause familial ALS is currently unknown. Two hypotheses have dominated recent discussion of the toxicity of ALS mutant CuZnSOD proteins: the oligomerization hypothesis and the oxidative damage hypothesis. The oligomerization hypothesis maintains that mutant CuZnSOD proteins are, or become, misfolded and consequently oligomerize into increasingly high-molecular-weight species that ultimately lead to the death of motor neurons. The oxidative damage hypothesis maintains that ALS mutant CuZnSOD proteins catalyze oxidative reactions that damage substrates critical for viability of the affected cells. This perspective reviews some of the properties of both wild-type and mutant CuZnSOD proteins, suggests how these properties may be relevant to these two hypotheses, and proposes that these two hypotheses are not necessarily mutually exclusive. PMID- 12655072 TI - Preorganization of molecular binding sites in designed diiron proteins. AB - De novo protein design provides an attractive approach to critically test the features that are required for metalloprotein structure and function. Previously we designed and crystallographically characterized an idealized dimeric model for the four-helix bundle class of diiron and dimanganese proteins [Dueferri 1 (DF1)]. Although the protein bound metal ions in the expected manner, access to its active site was blocked by large bulky hydrophobic residues. Subsequently, a substrate-access channel was introduced proximal to the metal-binding center, resulting in a protein with properties more closely resembling those of natural enzymes. Here we delineate the energetic and structural consequences associated with the introduction of these binding sites. To determine the extent to which the binding site was preorganized in the absence of metal ions, the apo structure of DF1 in solution was solved by NMR and compared with the crystal structure of the di-Zn(II) derivative. The overall fold of the apo protein was highly similar to that of the di-Zn(II) derivative, although there was a rotation of one of the helices. We also examined the thermodynamic consequences associated with building a small molecule-binding site within the protein. The protein exists in an equilibrium between folded dimers and unfolded monomers. DF1 is a highly stable protein (K(diss) = 0.001 fM), but the dissociation constant increases to 0.6 nM (deltadeltaG = 5.4 kcalmol monomer) as the active-site cavity is increased to accommodate small molecules. PMID- 12655073 TI - Why the kidney glomerulus does not clog: a gel permeation/diffusion hypothesis of renal function. AB - Current interpretations of kidney function in terms of a coarse filter followed by a fine filter have difficulty explaining why the glomerulus does not clog. I propose, as an alternative, a semiquantitative hypothesis that assumes that the size-selective property of the glomerulus is a consequence of the limited fraction of space in the glomerular basement membrane (a concentrated gel) into which macromolecules can permeate. The glomerular epithelial cell slits and slit diaphragms are assumed to impose substantial resistance to liquid flow across the glomerulus without acting as a molecular sieve. Calculations based on gel behavior show that proteins cross the glomerular basement membrane mainly by diffusion rather than by liquid flow, whereas water crosses entirely by flow. Thus, diffusion provides most of the protein, whereas flow provides the diluent. As a result, the single-nephron glomerular filtration rate (GFR) becomes a prime factor in (inversely) determining the concentration of proteins in early proximal tubular fluid. Because the reabsorption of proteins from the tubules is a saturable process, the gel permeationdiffusion hypothesis readily accounts for the albuminuria observed when single-nephron GFR is substantially reduced by severe pathological decreases in slit diaphragm length, such as occur in minimal change nephrotic syndrome in humans, in animals treated with puromycin aminonucleoside, or in humans or animals with mutations in the gene coding for nephrin. My hypothesis predicts that albuminuria will ensue, even with a normal kidney, if the single-nephron GFR falls below approximately 50% of normal. PMID- 12655074 TI - Enhanced oxygen diffusivity in interfaces of nanocrystalline ZrO2.Y2O3. AB - First measurements of oxygen grain boundary diffusion coefficients in nanocrystalline yttria-doped ZrO(2) (n-ZrO(2).6.9 mol % Y(2)O(3)) are presented. The (18)O diffusion profiles measured by secondary ion mass spectroscopy are much deeper in the nanocrystalline specimens than in single crystals. An oxygen diffusivity, D(B), in the grain boundaries can be deduced, which is approximately 3 orders of magnitude higher than in single crystals. From the present data the temperature variation of the oxygen grain boundary diffusivity, D(B) = 2.0 x 10( 5) exp (-0.91 eVk(B)T) m(2)s, and the oxygen surface exchange coefficient, k = 1.4 x 10(-2) exp (-1.13 eVk(B)T) ms, are derived. PMID- 12655075 TI - Do lipid rafts mediate virus assembly and pseudotyping? AB - Co-infection of a host cell by two unrelated enveloped viruses can lead to the production of pseudotypes: virions containing the genome of one virus but the envelope proteins of both viruses. The selection of components during virus assembly must therefore be flexible enough to allow the incorporation of unrelated viral membrane proteins, yet specific enough to exclude the bulk of host proteins. This apparent contradiction has been termed the pseudotypic paradox. There is mounting evidence that lipid rafts play a role in the assembly pathway of non-icosahedral, enveloped viruses. Viral components are concentrated initially in localized regions of the plasma membrane via their interaction with lipid raft domains. Lateral interactions of viral structural proteins amplify the changes in local lipid composition which in turn enhance the concentration of viral proteins in the rafts. An affinity for lipid rafts may be the common feature of enveloped virus proteins that leads to the formation of pseudotypes. PMID- 12655076 TI - Neutralizing epitopes specific for influenza B virus Yamagata group strains are in the 'loop'. AB - To study the neutralizing epitopes of influenza B virus Yamagata group strains, two monoclonal antibodies (mAbs) were used to select escape mutants of the virus. mAbs 5H4 and 3A12 were found to react with B/Yamagata group strains in haemagglutination inhibition and neutralization tests; no reactivity with B/Victoria group strains was observed. Most of the mutants reacted poorly to polyclonal ferret antibody against the 1998 isolate. Analysis of the deduced amino acid sequences identified a single amino acid substitution at residue 141 (Gly-->Arg) or 149 (Arg-->Gly) in 5H4-escape mutants and 141 (Gly-->Arg), 147 (Thr-->Ile) or 148 (Ser-->Gly) in 3A12-escape mutants. These residues are situated in close proximity in the 'loop' of the haemagglutinin molecule. These epitopes have been conserved in B/Yamagata group strains for almost 10 years in Japan but amino acid substitutions in the loop have been observed in clinical isolates only since 1999. PMID- 12655077 TI - Oral immunization with recombinant Yersinia enterocolitica expressing a measles virus CD4 T cell epitope protects against measles virus-induced encephalitis. AB - Immunization via the oral route with an attenuated Yersinia enterocolitica strain expressing a fragment of the measles virus nucleocapsid protein (aa 79-161) via its type III protein secretion system induced a T helper type 1 response in immunized C3H mice, which conferred protection against measles virus-induced encephalitis in a time- and dose-dependent manner. PMID- 12655078 TI - Recombinant Newcastle disease virus as a viral vector: effect of genomic location of foreign gene on gene expression and virus replication. AB - Newcastle disease virus (NDV) was examined for its suitability as a vector for the expression and delivery of foreign genes for vaccination and gene therapy. A reporter gene encoding human secreted alkaline phosphatase (SEAP) was inserted as an additional transcription unit at four different positions in the NDV genome, between the NP and P, M and F, and HN and L genes and behind the L gene. Eight infectious recombinant NDV (rNDV) viruses, four in the non-virulent strain NDFL and four in the virulent derivative NDFLtag, were generated by reverse genetics. SEAP expression levels, replication kinetics and virus yield were examined. Replication kinetics of the rNDV viruses in primary chicken embryo fibroblasts showed that the insertion of an additional gene resulted in a delay in the onset of replication. This effect was most prominent when the gene was inserted between the NP and P genes. With the exception of the strain that carried the SEAP gene behind the L gene, all recombinant strains expressed high levels of SEAP, both in cell culture and in embryonated chicken eggs. In embryonated eggs, the rNDV viruses showed a 2.6- to 5.6-fold (NDFL) or 2.1- to 8.1-fold (NDFLtag) reduction in yield compared with the parent strains. These results show that foreign genes can be inserted at different positions in the NDV genome without severely affecting replication efficiency or virus yield. PMID- 12655079 TI - Effects of a point mutation in the 3' end of the S genome segment of naturally occurring and engineered Bunyamwera viruses. AB - The genome of Bunyamwera virus (BUN) consists of three segments of single stranded RNA of negative polarity. The smallest segment, S, encodes the N protein and a nonstructural protein called NSs. We recently described a mutant virus (BUNdelNSs) that does not express NSs but overexpresses N and grows to lower titres than wild-type (wt) BUN. Here we report a BUNdelNSs variant that expresses lower levels of N protein and grows to higher titres. Sequencing of the 3' and 5' termini of the BUNdelNSs S RNA segment and analysis using a minireplicon system show that the N overexpressing phenotype results from a single nucleotide substitution at position 16 in the 3' terminus. This mutation could also be detected in wtBUN populations, and was isolated by plaquing a 'wt' variant carrying the mutation. This variant was found to express increased N and NSs levels, and grew to lower titres than wtBUN. PMID- 12655080 TI - Molecular epidemiology of rabies epizootics in Colombia: evidence for human and dog rabies associated with bats. AB - Three urban rabies outbreaks have been reported in Colombia during the last two decades, one of these is occurring in the Caribbean Region (northern Colombia), while the other two occurred almost simultaneously in Arauca (eastern Colombia) and in the Central Region and ended in 1997. In order to derive phylogenetic relationships between rabies viruses isolated in these three areas, 902 nt cDNA fragments encoding the cytoplasmic domain of protein G and a fragment of protein L were obtained by RT-PCR. These amplicons contained the G-L intergenic region and were sequenced to draw phylogenetic trees. Phylogenetic analysis showed three distinct groups of viruses in the study sample. Colombian genetic variant I viruses were isolated in both Arauca and the Central Region. These viruses are apparently extinct in Colombia. Colombian genetic variant II viruses were isolated in the Caribbean Region and are still being transmitted in that area. The third group of viruses consists of viruses isolated from two insectivorous bats, three domestic dogs and a human. According to sequence analysis, the data here indicate that the isolates in this third group are bat rabies virus variants. This finding is the first that associates bats to rabies in Colombian dogs and humans, showing an unsuspected vector threatening animal and public health. PMID- 12655081 TI - Phylogeography of infectious haematopoietic necrosis virus in North America. AB - Infectious hematopoietic necrosis virus (IHNV) is a rhabdoviral pathogen that infects wild and cultured salmonid fish throughout the Pacific Northwest of North America. IHNV causes severe epidemics in young fish and can cause disease or occur asymptomatically in adults. In a broad survey of 323 IHNV field isolates, sequence analysis of a 303 nucleotide variable region within the glycoprotein gene revealed a maximum nucleotide diversity of 8.6 %, indicating low genetic diversity overall for this virus. Phylogenetic analysis revealed three major virus genogroups, designated U, M and L, which varied in topography and geographical range. Intragenogroup genetic diversity measures indicated that the M genogroup had three- to fourfold more diversity than the other genogroups and suggested relatively rapid evolution of the M genogroup and stasis within the U genogroup. We speculate that factors influencing IHNV evolution may have included ocean migration ranges of their salmonid host populations and anthropogenic effects associated with fish culture. PMID- 12655083 TI - Sequencing of 'untypable' enteroviruses reveals two new types, EV-77 and EV-78, within human enterovirus type B and substitutions in the BC loop of the VP1 protein for known types. AB - The N-terminal part of VP1 was sequenced for 43 enterovirus isolates that could not initially be neutralized with LBM pools or in-house antisera. Most isolates were found to belong to human enterovirus type A (HEV-A) and HEV-B (18 isolates of each). All HEV-A isolates could be typed by sequencing, with CV (coxsackievirus)-A16 and EV (enterovirus)-71 being dominant (nine and seven isolates, respectively). These types thus seem to have diverged more from their prototypes than the other types. Among the HEV-B isolates, E-18 dominated with five isolates that became typable after filtration. The virus type obtained by molecular typing was verified for 28 of the other patient isolates by neutralization using high-titre monovalent antisera or LBM pools. Twenty-two of the other 30 'untypable' isolates had substitutions in the VP1 protein within or close to the BC loop. Two closely related HEV-B isolates diverged by 19.4 % from E-15, the most similar prototype. Two non-neutralizable HEV-C isolates split off from the CV-A13/CV-A18 branch, from which they diverged by 15.7-18.2 %. Three of the six non-neutralizable isolates, W553-130/99, W543-122/99 and W137-126/99, diverged by >24.2 % from the most similar prototype in the compared region. The complete VP1 was therefore sequenced and found to diverge by >29 % from all prototypes and by >28 % from each other. Strains similar to W553-130/99 that have been identified in the USA are tentatively designated EV-74. The two other isolates fulfil the molecular criterion for being new types. Since strains designated EV-75 and EV-76 have been identified in the USA, we have proposed the tentative designations EV-77 and EV-78 for these two new members of HEV-B. PMID- 12655082 TI - Amino acids 1-20 of the hepatitis C virus (HCV) core protein specifically inhibit HCV IRES-dependent translation in HepG2 cells, and inhibit both HCV IRES- and cap dependent translation in HuH7 and CV-1 cells. AB - A self-modulating mechanism by the hepatitis C virus (HCV) core protein has been suggested to influence the level of HCV replication, but current data on this subject are contradictory. We examined the effect of wild-type and mutated core protein on HCV IRES- and cap-dependent translation. The wild-type core protein was shown to inhibit both IRES- and cap-dependent translation in an in vitro system. This effect was duplicated in a dose-dependent manner with a synthetic peptide representing amino acids 1-20 of the HCV core protein. This peptide was able to bind to the HCV IRES as shown by a mobility shift assay. In contrast, a peptide derived from the hepatitis B virus (HBV) core protein that contained a similar proportion of basic residues was unable to inhibit translation or bind the HCV IRES. A recombinant vaccinia-HCV core virus was used to examine the effect of the HCV core protein on HCV IRES-dependent translation in cells and this was compared with the effects of an HBV core-recombinant vaccinia virus. In CV-1 and HuH7 cells, the HCV core protein inhibited translation directed by the IRES elements of HCV, encephalomyocarditis virus and classical swine fever virus as well as cap-dependent translation, whereas in HepG2 cells, only HCV IRES dependent translation was affected. Thus, the ability of the HCV core protein to selectively inhibit HCV IRES-dependent translation is cell-specific. N-terminal truncated (aa 1-20) HCV core protein that was expressed from a novel recombinant vaccinia virus in cells abrogated the inhibitory phenotype of the core protein in vivo, consistent with the above in vitro data. PMID- 12655084 TI - Molecular characterization of M1146, an American isolate of Ljungan virus (LV) reveals the presence of a new LV genotype. AB - Ljungan virus (LV) is a suspected human pathogen recently isolated from bank voles in Sweden. This study describes the genetic characterization of a virus, M1146, which was isolated in 1962 from another vole species (Microtus montanus), trapped in Oregon, USA. Based on antigenic properties, M1146 was postulated previously as a putative member of the family PICORNAVIRIDAE: The near complete genomic sequence verifies that M1146 is a member of the Picornaviridae, most closely related to LVs isolated in Sweden. The strain M1146 possesses typical LV genomic organization, including a cluster of two 2A homologues. There are significant differences throughout the capsid protein region, while the non structural region of M1146 is closely related to the Swedish LV genomes. Genetic and phylogenetic analyses show that M1146 represents a new genotype within the distinct LV cluster. Isolation of LV from both Swedish and American voles trapped over a period of 30 years suggests a continuous worldwide presence. PMID- 12655085 TI - Galactose is needed only for expression of co-receptors used by Theiler's murine encephalomyelitis virus as the virus does not directly bind galactose or use the UDP-galactose transporter as a receptor. AB - Theiler's murine encephalomyelitis virus (TMEV) infects most mammalian cells, but a TMEV receptor has not been identified. Studies have demonstrated that the UDP galactose transporter (UGT) is critical for TMEV attachment and entry into mammalian cells (Hertzler et al., Virology 286, 336-344, 2001). It was suggested that UGT might function as a TMEV receptor. We have demonstrated that polyclonal rabbit antibodies to human UGT that cross-react with hamster UGT do not block binding to or infection of mammalian cells by either high- or low-neurovirulence TMEV. In addition, incubation of virus with galactose, or blocking galactose on the cell surface with lectins, does not inhibit TMEV binding or infection. Thus, TMEV needs UGT for its transporter activity and galactose for assembly of its co receptors (attachment factors) but does not bind directly to galactose. Excluding direct involvement of UGT and galactose in TMEV binding and entry provides further insight into how TMEV interacts with the host cell and should facilitate ongoing studies to identify a TMEV receptor. PMID- 12655087 TI - Identification and analysis of gp116 and gp64 structural glycoproteins of yellow head nidovirus of Penaeus monodon shrimp. AB - Yellow head virus (YHV) is a major agent of disease in farmed penaeid shrimp. YHV virions purified from infected shrimp contain three major structural proteins of molecular mass 116 kDa (gp116), 64 kDa (gp64) and 20 kDa (p20). Two different staining methods indicated that the gp116 and gp64 proteins are glycosylated. Here we report the complete nucleotide sequence of ORF3, which encodes a polypeptide of 1666 amino acids with a calculated molecular mass of 185 713 Da (pI=6.68). Hydropathy analysis of the deduced ORF3 protein sequence identified six potential transmembrane helices and three ectodomains containing multiple sites for potential N-linked and O-linked glycosylation. N-terminal sequence analysis of mature gp116 and gp64 proteins indicated that each was derived from ORF3 by proteolytic cleavage of the polyprotein between residues Ala(228) and Thr(229), and Ala(1127) and Leu(1128), located at the C-terminal side of transmembrane helices 3 and 5, respectively. Comparison with the deduced ORF3 protein sequence of Australian gill-associated virus (GAV) indicated 83 % amino acid identity in gp64 and 71 % identity in gp116, which featured two significant sequence deletions near the N terminus. Database searches revealed no significant homology with other proteins. Recombinant gp64 expressed in E. coli with and without the C-terminal transmembrane region was shown to react with antibody raised against native gp64 purified from virions. PMID- 12655088 TI - A cytoplasmic region of the NSP4 enterotoxin of rotavirus is involved in retention in the endoplasmic reticulum. AB - The rotavirus genome encodes two glycoproteins, one structural (VP7) and one non structural (NSP4), both of which mature and remain in the endoplasmic reticulum (ER). While three amino acids in the N terminus have been proposed to function as a retention signal for VP7, no information is yet available on how NSP4 remains associated with the ER. In this study, we have investigated the ER retention motif of NSP4 by producing various C-terminal truncations. Deleting the C terminus by 52 amino acids did not change the intracellular distribution of NSP4, but an additional deletion of 38 amino acids diminished the ER retention and resulted in the expression of NSP4 on the cell surface. Brefeldin A treatment prevented NSP4 from reaching the cell surface, suggesting that C-terminal truncated plasma membrane NSP4 is transported through the normal secretory pathway. On the basis of these results, we propose that the region between amino acids 85 and 123 in the cytoplasmic region of NSP4 are involved in ER retention. PMID- 12655086 TI - Swine hepatitis E virus strains in Japan form four phylogenetic clusters comparable with those of Japanese isolates of human hepatitis E virus. AB - Japanese patients with sporadic acute hepatitis E are infected with polyphyletic strains of hepatitis E virus (HEV). Hepatitis E is considered a zoonotic disease. Thus far in Japan, only three strains of swine HEV have been identified and an antibody study for HEV antibodies has not been done on Japanese pigs. To determine the prevalence of swine HEV infection in Japan and the extent of genetic variation among Japanese swine HEV strains, we tested serum samples obtained from 2500 pigs from 2 to 6 months of age at 25 commercial swine farms in Japan for the presence of IgG antibodies to HEV and swine HEV RNA. Anti-HEV antibodies were detected in 1448 pigs (58 %). One-hundred-and-thirteen (15 %) of the 750 3-month-old pigs and 24 (13 %) of the 180 4-month-old pigs were positive for swine HEV RNA. The nucleotide sequence of a 412 bp region within open reading frame 2 of the 137 swine HEV isolates was determined. Sequence analyses revealed that the 137 isolates shared 76.6-100 % nucleotide sequence identities and were classifiable into genotype III (93 %) or IV (7 %) and that the isolates from the same farm were > or = 97.1 % similar to each other. Phylogenetic analysis showed that the Japanese swine and human HEV isolates segregated into four clusters, with the highest nucleotide identity being 94.4-100 % between swine and human isolates in each cluster. These results indicate that swine HEV is widespread in the Japanese swine population and further support the hypothesis that swine serve as reservoirs for HEV infection. PMID- 12655089 TI - Network analysis of human and simian immunodeficiency virus sequence sets reveals massive recombination resulting in shorter pathways. AB - The intrinsic recombination rate of human immunodeficiency virus (HIV) exceeds the point mutation rate by a factor of 10. As the majority of infected cells in vivo harbour multiple proviruses, the stage is set for rampant recombination. Therefore, it may be presumed that phylogenic relationships and mutation frequencies will probably be affected by recombination. However, the proportion of homoplasies arising from recombination and mutation is not known. By studying the evolution of the hypervariable regions of the simian immunodeficiency virus envelope gene among four macaques, it is shown that homoplasies arise more from recombination than from point mutation. When recombination is accounted for, the minimum number of substitutions in a sequence set may be reduced by as much as 45 %. In fact, the true number of point mutations in a set of HIV sequences tends to the number of discrete substitutions. Hence, lineages are younger than anticipated previously, although not in proportion to the ratio of the intrinsic recombination/point mutation rate. Recombination also inflates codon polymorphisms. PMID- 12655090 TI - P53 facilitates degradation of human T-cell leukaemia virus type I Tax-binding protein through a proteasome-dependent pathway. AB - Human T-cell leukaemia virus type 1 (HTLV-I), the aetiological agent of adult T cell leukaemia (ATL) and tropical spastic paraparesis (TSP/HAM), transforms human T-cells in vivo and in vitro. The Tax protein of HTLV-I is essential for cellular transformation as well as viral and cellular gene transactivation. The interaction of Tax with cellular proteins is critical for these functions. We previously isolated and characterized a novel Tax-binding protein, TRX (TAX1BP2), by screening a Jurkat T-cell cDNA library. In the present study, we present evidence that the tumour suppressor p53 targets the TRX protein for proteasome degradation. Pulse-chase experiments revealed that p53 enhanced the degradation of TRX protein and reduced the half-life from 2.0 to 0.25 h. p53 mutants R248W and R273H enhance TRX degradation suggesting a transcriptionally independent mechanism. Both HTLV-I Tax and the proteasome-specific inhibitor MG132 inhibited p53-mediated TRX protein degradation. These results suggest that TRX degradation is mediated through activation of the proteasome protein degradation pathway independent of transcriptional function of p53. Our results provide the first experimental evidence that Tax inhibits transcription-dependent and independent functions of p53. PMID- 12655092 TI - Divergence of reiterated sequences in a series of genital isolates of herpes simplex virus type 1 from individual patients. AB - Both serotypes of herpes simplex virus (HSV), HSV-1 and HSV-2, are aetiological agents of genital herpes, although genital herpes caused by HSV-1 recurs less frequently. The HSV-1 genome contains a number of short, tandemly repeated sequences, and some reiterated sequences can serve as sensitive markers for the differentiation of HSV-1 strains. In the present study, variation in reiterations (assumed to be due to different copy numbers of tandemly repeated sequences) was examined in HSV-1 isolates from genital lesions from the same individual. Six sets (three primary-recurrence sets and three multiple-recurrence sets) of HSV-1 isolates were analysed: the primary-recurrence set consisted of two isolates (one isolated at a primary episode and the other at a recurrent episode) from the same individual; the multiple-recurrence set consisted of plural isolates from different episodes of recurrence in the same individual. Variations in length of the major DNA fragment, containing reiteration I (within the a sequence) and/or reiteration IV (within introns of genes US1 and US12), were detected between isolates of each multiple-recurrence set, but not of the primary-recurrence set. Thus, HSV-1 isolates of multiple-recurrence sets are assumed to have diverged more widely within each set than those of primary-recurrence sets, probably because of more rounds of virus DNA replication. This divergence of reiterations seems to indicate a forward step in the division of HSV-1 from a common ancestor into different lineages. PMID- 12655091 TI - A glucocorticoid response element in the LTR U3 region of Friend murine leukaemia virus variant FIS-2 enhances virus production in vitro and is a major determinant for sex differences in susceptibility to FIS-2 infection in vivo. AB - The nucleotide sequence of the Friend murine leukaemia virus variant FIS-2 LTR has high identity with the closely related Friend murine leukaemia virus (F-MuLV) LTR, except for the deletion of one direct repeat, a few point mutations and the generation of a glucocorticoid response element (GRE) in the U3 region. The GRE can mediate gene induction by glucocorticoids, mineral corticoids, progesterone and androgens, and it has been shown that incorporation of a GRE(s) within the LTR can increase the transcriptional activity of retroviral enhancers. We have previously reported an increased early virus replication in male mice compared with female mice when infected with a virus containing the FIS-2 LTR and have proposed that the GRE might contribute to this sex difference. In the present study, we introduced a single point mutation in the GRE and performed comparative studies in NIH 3T3 cells and in young adult male and female NMRI mice. We found that significantly more virus was produced from NIH 3T3 cells infected with wt FIS-2 than from cells infected with the FIS-2 GRE mutant and that this difference was further augmented by glucocorticoids. The glucocorticoid antagonist RU486 inhibited virus production in a dose-dependent manner. The wt FIS-2 disseminated significantly faster than the FIS-2 GRE mutant in both male and female mice. There was no significant difference in the dissemination rate between male and female mice infected with the FIS-2 GRE mutant. Hence, the GRE in the FIS-2 LTR is one determinant of the significant sex difference in susceptibility to FIS-2 infection. PMID- 12655093 TI - RNase L activity does not contribute to host RNA degradation induced by herpes simplex virus infection. AB - In early herpes simplex virus (HSV) infection, the virion host shutoff (vhs) protein mediates the degradation of mRNA and subsequent shutoff of host protein synthesis. It is unclear whether vhs acts alone or in concert with virus-induced cellular factors for this activity. This paper examines whether RNase L, a virally induced endoribonuclease, contributes to HSV-induced mRNA decay. Results showed that RNA degradation was comparable in wild-type and RNase L(-/-) cells, demonstrating that HSV-mediated RNA degradation is independent of RNase L activity. Furthermore, the data show that HSV-1 does not significantly induce RNase L activity in murine embryo fibroblasts. PMID- 12655094 TI - Stimulation of bovine herpesvirus-1 productive infection by the adenovirus E1A gene and a cell cycle regulatory gene, E2F-4. AB - Identifying cellular genes that promote bovine herpesvirus-1 (BHV-1) productive infection is important, as BHV-1 is a significant bovine pathogen. Previous studies demonstrated that BHV-1 DNA is not very infectious unless cotransfected with a plasmid expressing bICP0, a viral protein that stimulates expression of all classes of viral promoters. Based on these and other studies, we hypothesize that the ability of bICP0 to interact with and modify the function of cellular proteins stimulates virus transcription. If this prediction is correct, cellular proteins that activate virus transcription could, in part, substitute for bICP0 functions. The adenovirus E1A gene and bICP0 encode proteins that are potent activators of viral gene expression, they do not specifically bind DNA and both proteins interact with chromatin-remodelling enzymes. Because of these functional similarities, E1A was tested initially to see if it could stimulate BHV-1 productive infection. E1A consistently stimulates BHV-1 productive infection, but not as efficiently as bICP0. The ability of E1A to bind Rb family members plays a role in stimulating productive infection, suggesting that E2F family members activate productive infection. E2F-4, but not E2F-1, E2F-2 or E2F-5, activates productive infection with similar efficiency as E1A. Next, E2F family members were examined for their ability to activate the BHV-1 immediate-early (IE) transcription unit 1 (IEtu1) promoter, as it regulates IE expression of bICP0 and bICP4. E2F-1 and E2F-2 strongly activate the IEtu1 promoter, but not a BHV-1 IEtu2 promoter or a herpes simplex virus type 1 ICP0 promoter construct. These studies suggest that E2F family members can stimulate BHV-1 productive infection. PMID- 12655095 TI - Antibody-induced internalization of viral glycoproteins and gE-gI Fc receptor activity protect pseudorabies virus-infected monocytes from efficient complement mediated lysis. AB - Pseudorabies virus (PRV)-infected blood monocytes are able to transport virus throughout the body of vaccination-immune pigs. PRV-infected monocytes express viral glycoproteins in their plasma membrane that can be recognized by virus specific antibodies. Recently, it has been shown that addition of PRV-specific polyclonal immunoglobulins to PRV-infected monocytes at 37 degrees C induces internalization of the majority of plasma membrane-expressed viral glycoproteins. This study investigated whether this process may interfere with efficient antibody-dependent complement-mediated lysis (ADCML) of infected monocytes. Therefore, an ADCML assay was set up in vitro. A significant decrease in the percentage of cells lysed by ADCML was observed when antibody-induced internalization of PRV glycoproteins occurred (P<0.005). Furthermore, it is shown (i) that the PRV gE-gI complex, which, like certain other alpha herpesvirus orthologues, possesses IgG-binding capacity, aids in avoiding efficient ADCML of PRV-infected monocytes and (ii) that the efficiency of PRV gE-gI-mediated evasion of ADCML can be decreased by the presence of gE-gI-specific antibodies. PMID- 12655096 TI - Heterogeneous Epstein-Barr virus latent gene expression in AIDS-associated lymphomas and in type I Burkitt's lymphoma cell lines. AB - Epstein-Barr virus (EBV) is associated with lymphoma in immunocompromised patients. This study provides evidence that the expression of EBV nuclear antigen 3 genes can be directed from the F promoter in different type I Burkitt's lymphoma cell lines and in some lymphomas from human immunodeficiency virus infected patients. This expression occurs predominantly after induction of the EBV lytic cycle. PMID- 12655097 TI - The BZLF1 promoter of Epstein-Barr virus is controlled by E box-/HI-motif-binding factors during virus latency. AB - The BZLF1 open reading frame of Epstein-Barr virus (EBV) encodes an important transactivator of replication. During latency, transcription of this gene is switched off. HI motifs have been shown to cause negative regulation of the promoter. Using yeast one-hybrid assays, we isolated the E box-binding protein, E2-2, interacting with these motifs. Electrophoretic mobility shift assays demonstrated that E2-2 binds to HI alpha, HI beta and HI gamma, which contain E box consensus binding sites. Deletion of the HI-associated E boxes and overexpression of E2-2 in transfection assays revealed that these elements act as repressors in lymphoid cells. In contrast, in epithelial cells they contribute to the increased responsiveness of the promoter to transactivation by the BZLF1 protein. The data presented are in accord with an alternative and exclusive binding of different cell type- and differentiation-specific factors, such as E2 2, to the HI-associated E boxes in lymphoid and epithelial cells. This implies a role in cell type-specific virus replication. PMID- 12655099 TI - Double-stranded RNA-binding proteins could suppress RNA interference-mediated antiviral defences. AB - RNA interference (RNAi) is a double-stranded (ds)RNA-inducible, sequence-specific RNA-degradation mechanism that operates as a natural antiviral system in plants and animals. Successful virus infection requires evasion or suppression of RNAi. Indeed, RNAi suppressor proteins have been identified in plant and animal viruses, although the molecular mechanism of silencing inhibition is still poorly understood. Because many RNA viruses encode dsRNA-binding proteins (dsRBPs) and as RNAi is triggered by the accumulation of dsRNAs, dsRBPs were examined to see if they inhibit RNAi. Here, it is shown that heterologous dsRBPs suppressed RNAi in plants, indicating that in natural host-virus interactions, pathogen-encoded dsRBPs could inactivate RNAi-mediated host defences. PMID- 12655098 TI - A novel function for the Epstein-Barr virus transcription factor EB1/Zta: induction of transcription of the hIL-10 gene. AB - Interleukin-10 (IL-10) plays a critical role in Epstein-Barr virus (EBV) biology. Indeed, the EBV genome contains a gene (BCRF1) with homology to the human IL-10 (hIL-10) gene. In addition to viral IL-10, which is secreted late in the productive cycle, hIL-10 production is also induced in B cells infected by EBV. The EBV protein LMP-1 and the viral small non-polyadenylated RNAs (EBERs) expressed during latency are involved in hIL-10 induction. In this study, we show that in B cells the viral transcription factor EB1, which is the main inducer of the EBV productive cycle, also activates transcription of the hIL-10 gene and secretion of the hIL-10 protein. Accordingly, EB1 bound directly to specific DNA sequences in the hIL-10 minimal promoter. Moreover, specific disruption of EB1 binding to some of these sites impaired EB1-mediated activation of transcription at the hIL-10 promoter in a transient expression assay. Therefore, an increase in IL-10 production occurs during latency and early and late during the productive cycle. This production of IL-10 might favour the survival of EBV-infected cells in vivo and/or create a microenvironment required for efficient de novo infection of B lymphocytes by EBV virions. PMID- 12655100 TI - Assembly of single-shelled cores and double-shelled virus-like particles after baculovirus expression of major structural proteins P3, P7 and P8 of Rice dwarf virus. AB - Expression of the core capsid protein P3 of Rice dwarf virus in a baculovirus system resulted in the formation of single-shelled core-like particles in insect cells in the absence of any other capsid proteins. Double-shelled virus-like particles were also observed upon mixing or co-expression of P3 and the major outer capsid protein P8, suggesting that P3 and P8 have the ability to form double-shelled particles both in vivo and in vitro. Core protein P7 expressed in a similar manner was incorporated into the virus-like particles. PMID- 12655101 TI - Immunodetection and fluorescent microscopy of transgenically expressed hordeivirus TGBp3 movement protein reveals its association with endoplasmic reticulum elements in close proximity to plasmodesmata. AB - The subcellular localization of the hydrophobic TGBp3 protein of Poa semilatent virus (PSLV, genus Hordeivirus) was studied in transgenic plants using fluorescent microscopy to detect green fluorescent protein (GFP)-tagged protein and immunodetection with monoclonal antibodies (mAbs) raised against the GFP based fusion expressed in E. coli. In Western blot analysis, mAbs efficiently recognized the wild-type and GFP-fused PSLV TGBp3 proteins expressed in transgenic Nicotiana benthamiana, but failed to detect TGBp3 in hordeivirus infected plants. It was found that PSLV TGBp3 and GFP-TGBp3 had a tendency to form large protein complexes of an unknown nature. Fractionation studies revealed that TGBp3 represented an integral membrane protein and probably co-localized with an endoplasmic reticulum-derived domain. Microscopy of epidermal cells in transgenic plants demonstrated that GFP-TGBp3 localized to cell wall-associated punctate bodies, which often formed pairs of opposing discrete structures that co localized with callose, indicating their association with the plasmodesmata enriched cell wall fields. After mannitol-induced plasmolysis of the leaf epidermal cells in the transgenic plants, TGBp3 appeared within the cytoplasm and not at cell walls. Although TGBp3-induced bodies were normally static, most of them became motile after plasmolysis and displayed stochastic motion in the cytoplasm. PMID- 12655102 TI - The optimal temperature for RNA replication in cells infected by Soil-borne wheat mosaic virus is 17 degrees C. AB - Systemic infection of wheat plants with Soil-borne wheat mosaic virus (SBWMV) requires temperatures below 20 degrees C. Here we examine the cause of the temperature sensitivity by inoculating infectious in vitro transcripts of SBWMV RNA1 and RNA2 to barley mesophyll protoplasts. After RNA inoculation, protoplasts were incubated at temperatures between 15 and 25 degrees C for up to 48 h. Western blot analysis showed that the capsid protein accumulated most abundantly at 17 degrees C but was not detectable at 25 degrees C. Northern blot analysis showed that the wild-type RNA1 and RNA2 and their subgenomic RNAs accumulated most abundantly at 17 degrees C but were barely detectable at 25 degrees C. An RNA1 mutant in which the p152 and p211 replicase genes were placed between the 5' and 3'-untranslated regions also replicated most efficiently at 17 degrees C but not at 25 degrees C. Thus, the requirement for temperatures lower than 20 degrees C for SBWMV infection is primarily determined by replication of RNA1, which encodes the viral RNA replicase. PMID- 12655103 TI - Potato mop-top virus: the coat protein-encoding RNA and the gene for cysteine rich protein are dispensable for systemic virus movement in Nicotiana benthamiana. AB - Full-length genomic cDNA clones of the Swedish isolate of Potato mop-top virus (PMTV) were transcribed in vitro using T7 RNA polymerase. The combination of RNA 1, 2 and 3 synthesized in the presence of m(7)GpppG cap analogue was infectious when inoculated onto Nicotiana benthamiana plants. Also, the combination of RNA 1 (encodes the viral replicase) with RNA 3 [encodes the triple gene block proteins and a small cysteine-rich protein (CRP)] was infectious and both RNAs moved systemically in N. benthamiana plants in the absence of RNA 2, which encodes the coat protein (CP). However, the yellow mosaic symptoms that typically developed following PMTV infection with all three RNAs were not observed in plants infected with RNA 1+RNA 3. Site-directed mutagenesis experiments revealed that expression of the putative CRP was not required for systemic infection and symptom induction in N. benthamiana. These data show that PMTV represents an example of a multipartite virus capable of establishing systemic infection without the CP encoding RNA, and also without the putative CRP. PMID- 12655104 TI - Nucleotide sequence and genome organization of Cucumber yellows virus, a member of the genus Crinivirus. AB - The genome of Cucumber yellows virus (CuYV), isolated in Japan from cucumber (Cucumis sativus L.), was completely sequenced and shown to be bipartite. CuYV RNA1 consisted of 7889 nucleotides and encompassed seven open reading frames (ORFs), which is typical of the Closteroviridae, including a heat-shock protein 70 homologue, a coat protein and a diverged coat protein (CPd). CuYV RNA2 consisted of 7607 nucleotides and included two ORFs: ORF1a potentially encoded a polyprotein containing putative papain-like protease, methyltransferase and helicase domains, and ORF 1b potentially encoded an RNA-dependent RNA polymerase, which is probably expressed via a +1 ribosomal frameshift. The size and organization of the CuYV genome are similar to those of Lettuce infectious yellows virus (LIYV), the type member of the genus Crinivirus in the family Closteroviridae, indicating that CuYV is a member of that genus, although CuYV differed in several points from LIYV. PMID- 12655105 TI - In vitro cell-free conversion of bacterial recombinant PrP to PrPres as a model for conversion. AB - Prion diseases are associated with the conversion of the normal cellular prion protein, PrP(C), to the abnormal disease-associated protein, PrP(Sc). This conversion can be mimicked in vitro using PrP(Sc) isolated from the brains of scrapie-infected animals to induce conversion of recombinant PrP(C) into a proteinase K-resistant isoform, PrP(res). Traditionally, the 'cell-free' conversion assay has used, as substrate, recombinant PrP(C) purified from mammalian tissue culture cells or, more recently, from baculovirus-infected insect cells. The cell-free conversion assay has been modified by replacing the tissue culture-derived PrP(C) with recombinant PrP purified from bacteria. Bacterial expression and chromatographic purification give high yields of recombinant radiolabelled untagged protein, eliminates artefacts that may be due to cellular factors or antibody fragments normally present in labelled PrP preparations and allows accurate and rapid variation of protein sequence using standard molecular biological techniques. In addition, these cell-free conversion assays were carried out under more physiological conditions, giving more relevance to the assay as a model for conversion. To validate its use in this assay, this bacterial recombinant PrP has been shown to have the conversion properties of mammalian PrP(C): (i) it converts to a proteinase K-resistant isoform in the presence of PrP(Sc); (ii) the efficiency of this conversion by PrP(Sc) of different strains and species parallels that found in vivo; and (iii) its cell-free conversion is inhibited by Congo Red analogues in a structure dependent manner similar to that seen in in vivo and in vitro cell assays. PMID- 12655106 TI - Studies of the transmissibility of the agent of bovine spongiform encephalopathy to pigs. AB - Studies to test the transmissibility of the bovine spongiform encephalopathy (BSE) agent to pigs began in 1989. Parenteral inoculation of the agent by three routes simultaneously (intracranially, intravenously and intraperitoneally) produced disease with an incubation period range of 69-150 weeks. Pre-clinical pathological changes were detected in two pigs killed electively at 105 and 106 weeks post-inoculation. Infectivity was detected by bioassay in inbred mice in the CNS of those pigs that developed spongiform encephalopathy. Infectivity was also found in the stomach, jejunum, distal ileum and pancreas of terminally affected pigs. These findings show that pigs are susceptible to BSE. In contrast, disease failed to occur in pigs retained for 7 years after exposure by feeding BSE-affected brain on three separate days, at 1-2 week intervals. The amounts fed each day were equivalent to the maximum daily intake of meat and bone meal in rations for pigs aged 8 weeks. No infectivity was found in tissues assayed from the pigs exposed orally. This included tissues of the alimentary tract. It is suggested that these pigs did not become infected. The relatively high oral exposure used in these experiments compared with feed-borne exposure in the field may explain the absence of an epidemic of spongiform encephalopathy in domestic pigs concurrent with the BSE epidemic in the UK. PMID- 12655107 TI - Cell-associated variants of disease-specific prion protein immunolabelling are found in different sources of sheep transmissible spongiform encephalopathy. AB - Scrapie and bovine spongiform encephalopathy (BSE) are transmissible spongiform encephalopathies (TSEs) or prion diseases affecting domestic and exotic ruminants. In previous immunohistochemical studies, we have shown that different sheep TSE sources may be distinguished by both the proportion of disease-specific prion protein (PrP(d)) accumulation relative to different cell types in the brain (the 'PrP(d) profile') and by different labelling patterns for PrP peptide sequences within phagocytic cells. In the present study, we have further characterized the intracellular accumulation patterns of PrP(d) in the lymphoreticular system (LRS) and in the brain of sheep clinically affected with scrapie or BSE. BSE-infected PrP(ARQ/ARQ) sheep of different breeds were compared with scrapie-infected sheep of different PrP genotypes. Cases of BSE infection could be distinguished from scrapie cases by a marked reduction in labelling of PrP(d) containing the 84-105 amino acid residues in phagocytic cells of the LRS and in neurones and glia of the brain. These results therefore indicate that TSE agent-dependent processing of PrP in specific cell types within the brain and LRS can be used to distinguish between BSE in PrP(ARQ/ARQ) sheep and scrapie in sheep of several PrP genotypes. Three different N-terminal peptide antibody labelling patterns were recognized for different cell types in different tissues of BSE infected sheep, suggesting that different truncated forms of PrP(d) are formed following infections with this agent strain. These variations in the cleavage sites of BSE PrP(d) may be due to cell-specific variation in endosomal-lysosomal digestion or to cell- and tissue-specific differences in BSE PrP(d) conformation. PMID- 12655108 TI - Molecular analysis of iatrogenic scrapie in Italy. AB - An accidental intra- and interspecies transmission of scrapie occurred in Italy in 1997 and 1998 following exposure to a vaccine against Mycoplasma agalactiae. PrP(Sc) in affected sheep and goats, collected from a single flock exposed to vaccination 2 years earlier, was molecularly typed. In five animals with iatrogenic scrapie, a PrP(Sc) type with a 20 kDa core fragment was found in all areas of the brain investigated. In three sheep and one goat, this isoform co occurred with a fully glycosylated isoform that had a protease-resistant backbone of 17 kDa, whereas in two sheep and four goats, the two PrP(Sc) types were detected in different regions of the brain. In sheep with natural field scrapie, a PrP(Sc) type with physico-chemical properties indistinguishable from the 20 kDa isoform was found. The present results suggest the co-presence of two prion strains in mammary gland and brain homogenates used for vaccination. PMID- 12655109 TI - Campylobacter spp in human, chickens, pigs and their antimicrobial resistance. AB - Campylobacter spp. have been identified as etiologic agents in outbreaks and sporadic cases of gastroenteritis in developed countries. In developing countries, most reported Campylobacter infections are in children. Previously reported prevalences of Campylobacter spp. in children in Southeast Asia range from 2.9% to 15%. The frequency and pattern of occurrence of Campylobacter spp. differ between developed and developing countries, especially in the number of cases reported in adults and the presence of any seasonal patterns in occurrence. Although the severity of Campylobacter infection in adults was different between developed and developing countries, the clinical symptoms of infection in adults resulting from infection in developing countries was similar to those in developed countries. Many different animal species maintain Campylobacter spp. with no clinical signs. There do not appear to be significantly different colonization rates of Campylobacter in food animals between developed and developing countries. The role of C. jejuni as a primary pathogen in farm animals is uncertain. C. jejuni can be found in feces of diarrheic and healthy calves and piglets. Campylobacter with resistance to antimicrobial agents have been reported in both developed and developing countries, and the situation seems to deteriorate more rapidly in developing countries, where there is widespread and uncontrolled use of antibiotics resistance was observed at high levels in food animals in both developed and developing countries. Studies suggested an association between antimicrobial use in food animals and the development of resistance in human isolates in developed countries. PMID- 12655110 TI - The cellular differentiation of M cells from crypt undifferentiated epithelial cells into microvillous epithelial cells in follicle-associated epithelia of chicken cecal tonsils. AB - To clarify the cellular origin and the fate of M cells, detailed distributions of the epithelial cells were investigated scanning electron microscopically on the follicle-associated epithelia (FAE) of chicken cecal tonsils. The distribution of M cells was closely related with the situation of the crypt orifices in chicken cecal tonsils. In undeveloped cecal tonsils, the intestinal crypts were localized at the periphery of the FAE. In these tonsils, M cells without microvilli (M(0)) were predominantly populated in the basal region of the FAE, whereas goblet cells and microvillous epithelial cells (MV) were more distributed in the middle to the apical region of the FAE. A few M cells with short microvilli were dispersed throughout the FAE. Significantly shrunk MV (MVs) clustered together in transitional portions from the lateral face to the roof of the FAE. In well developed cecal tonsils, the crypts also opened at the lateral surface in addition to the periphery of the FAE. In these tonsils, the M(0) accumulated densely in the small areas around the crypt orifices exclusively. No sign of exfoliation of apoptotic epithelial cells was found in the M(0)-accumulated areas and at their peripheral boundaries. The MVs were often clustered in the central regions among the crypt orifices in addition to the roof of the FAE. These findings suggest that M cells are directly derived from the undifferentiated crypt epithelial cells, not fall into apoptotic cell death and further differentiate into MV in the FAE of chicken cecal tonsils. PMID- 12655111 TI - Age-related histological changes in the canine substantia nigra. AB - Age-related changes in the canine substantia nigra, were examined using immunostaining for tyrosine hydroxylase (TH), glial fibrillary acidic protein (GFAP), neurofilament (NF), ubiquitin, single stranded DNA (ssDNA), and alpha synuclein (alphaSN). Brain sections from 34 necropsied dogs, ranging from 2 months to 18 years old, were used for this study. On general histological examinations, several age-related changes, including lipofuscin deposition, polyglucosan bodies, amorphous basophilic inclusions and eosinophilic crystal inclusions, were found in the aged dogs. Immunohistochemically, TH-positive neurons were located only in the substantia nigra. The number of TH-positive neurons was well preserved in all dogs examined, however, the ratio of TH positive neurons to GFAP-positive glial cells tended to show slight decrease in aged dogs. By ssDNA immunostaining for apoptotic cells, there were no significant results in the number of ssDNA-positive neurons. The number of ubiquitin- and NF positive swollen neurites was increased markedly in aged dogs. Ubiquitin immunostaining revealed a small number of basophilic and eosinophilic inclusions, although both types of inclusions were negative for NF. By alphaSN immunostaining, no neurons were immunoreactive and no basophilic or eosinophilic intracytoplasmic inclusions were revealed. These results indicate that in the substantia nigra of aged dogs the dopaminergic neurons are well preserved, but intracytoplasmic inclusions and ubiquitin-positive degenerative neurites are commonly found. PMID- 12655112 TI - Effects of Bacillus thuringiensis Cry1Ab toxin on mammalian cells. AB - The Cry proteins produced by Bacillus thuringiensis are considered to be highly specific insecticidal proteins. Judged to be safe for humans and farm animals due to their insect-oriented selective toxicity, the proteins have been utilized as a biological pesticide and introduced into genetically modified plants. However, some critical fundamental characters of the Cry proteins remain unclear, and the direct effects of activated Cry proteins on mammalian cells have not yet been fully confirmed. Therefore, in this study we employed primary cultured bovine hapatocytes as a model system to determine if Cry1Ab, a Cry protein, affects mammalian cells. There were no significant changes in the secretion of albumin or the morphology of the Cry1Ab-treated cells. The LDH release showed a tendency to increase after the administration of Cry1Ab, but not significantly. Taking these results on bovine hepatocytes into consideration, Cry1Ab has little acute toxicity on mammalian cells. PMID- 12655113 TI - Relation between reticulocyte count and characteristics of erythrocyte 5' nucleotidase in dogs, cats, cattle and humans. AB - To examine substrate specificity and susceptibility to lead, erythrocyte 5' nucleotidase was measured in dogs, cats, cattle and humans, and its relationship to the reticulocyte count in these species was determined. The reticulocyte count in dogs was similar to that in humans, but the count in cats was higher than that in humans. Reticulocytes were not observed in cattle. The activities of canine erythrocyte 5'-nucleotidase measured using cytidine and uridine 5' monophosphates, which are preferentially catalyzed by one of the human pyrimidine 5'-nucleotidase isozymes (P5N-I), were similar to those of the human enzyme. The canine enzyme preferentially catalyzed thymidine 3'-monophosphate, which is catalyzed only by human P5N-II, more strongly than the human enzyme. This suggests that canine erythrocytes have two isozymes similar to human P5N-I and P5N-II, and a higher P5N-II-like activity than human erythrocytes. Feline erythrocytes had the highest level of P5N-I-like activity among the species examined, and the bovine enzymic activities including those of P5N-I and II were the lowest among these species. According to these observations, the reticulocyte count was approximately proportional to the P5N-I-like activity in these species. Therefore, the P5N-I-like activity may be involved in the morphological maturation of mammalian erythrocytes. The canine and feline erythrocytes had markedly high activity and preferentially catalyzed purine 5'-monophosphate suggesting the presence of a purine-specific 5'-nucleotidase as in human erythrocytes. In addition, the canine and feline P5N-I-like activity showed less susceptibility to lead than the human P5N-I. This may be a reason why there are few case reports of lead-induced anemia in dogs and cats. PMID- 12655114 TI - Bovine apolipoprotein E in plasma: increase of ApoE concentration induced by fasting and distribution in lipoprotein fractions. AB - Apolipoprotein E (apoE) is a protein constituent of lipoproteins, and acts as a receptor-binding ligand. Although the existence of bovine apoE in lipoprotein fractions has already been reported, quantitative studies on the changes of apoE in plasma and lipoprotein fractions are lacking. In the present study, an increase of a 38 kDa protein in the very low-density lipoprotein (VLDL) fraction obtained from fasted calves was detected. This 38 kDa protein was identified as bovine apoE by determination of the N-terminal amino acid sequence. Bovine apoE was purified and an enzyme-linked immunosorbent assay (ELISA) was developed. Using this system, the effect of fasting on the concentration of apoE in plasma and the distribution of apoE in lipoprotein fractions were investigated. After 3 days of fasting, the concentration of plasma apoE increased significantly (p<0.05) by 280 %, and was returned to the basal level by 3 days of refeeding. The lipoprotein fractions obtained from before and after fasting was separated by ultracentrifugation. ApoE was significantly increased in VLDL, low-density lipoprotein (LDL) and non-lipoprotein fractions by fasting (p<0.05). On the other hand, in high-density lipoprotein (HDL) fractions obtained from both before and after fasting, the level of apoE was very low compared to the other fractions. These results suggested that bovine apoE contents in triglyceride-rich lipoproteins are modulated by nutritional treatment and closely associated with triglyceride-rich lipoprotein metabolism. PMID- 12655116 TI - Clinical and clinico-pathologic characteristics of Shiba dogs with a deficiency of lysosomal acid beta-galactosidase: a canine model of human GM1 gangliosidosis. AB - The present study was conducted to determine the clinical and clinico-pathologic characteristics of Shiba dogs with GM1 gangliosidosis, which is due to an autosomal recessively inherited deficiency of lysosomal acid beta-galactosidase activity. Clinical and clinico-pathological features were investigated in 10 homozygous Shiba dogs with GM1 gangliosidosis. The age at onset was 5 to 6 months and the dogs manifested progressive neurologic signs including loss of balance, intermittent lameness, ataxia, dysmetria and intention tremor of the head. The dogs were unable to stand by 10 months of age due to a progression of ataxia and spasticity in all limbs. Corneal clouding, a visual defect, generalized muscle rigospasticity, emotional disorder and a tendency to be lethargic were observed at 9 to 12 months. The dogs became lethargic from 13 months of age. The survival period seemed to be 14 to 15 months. As a clinico-pathologic feature, lymphocytes with abnormally large vacuoles were observed in peripheral blood (30 to 50% of total lymphocytes) through the lifetime of the dogs. The clinical and clinico pathologic characteristics of this animal model are useful for not only the development and testing of potential methods of therapy, but also the diagnosis of affected homozygous Shiba dogs in veterinary clinics. PMID- 12655115 TI - Flow cytometric assessment of ethylene glycol monoethyl ether on spermatogenesis in rats. AB - The effects of ethylene glycol monoethyl ether (EGEE) on testicular cell populations in rats were investigated by a flow cytometric method. Rats were administered by gavage with EGEE at the various doses of 0 (saline alone), 100, 200, 400, and 800 mg/kg body weight/day for 4 weeks. The treatment of EGEE caused decreases in the weight of testis and epididymis and in the number of testicular cells. Histopathologically, exfoliation of germ cells into the tubular lumen was observed at the doses of above 200 mg/kg. The treatment of EGEE at the dose of 400 mg/kg caused moderate testicular degeneration. A significant depletion of haploid cells and a disproportionate ratio of diploid and tetraploid cells were observed as determined by flow cytometric analysis. These results indicate that the toxic effect of EGEE on the male reproductive system may be strongly associated with the disproportion of testicular germ cells. PMID- 12655117 TI - Large-scale production of porcine mature interleukin-18 (IL-18) in silkworms using a hybrid baculovirus expression system. AB - In this report, a hybrid baculovirus expression system, which means a hybrid virus of the Autographa californica nuclear polyhedrosis virus and the Bombyx mori nuclear polyhedrosis virus, was used for the large-scale production of porcine mature interleukin-18 (IL-18) in silkworms. Two recombinant hybrid baculoviruses containing cDNA of the porcine precursor IL-18 and the porcine caspase-1 were constructed and were used to infect silkworm larvae. After the co infection of the two viruses, porcine mature IL-18 was efficiently produced in the haemolymph. The concentration of IL-18 in the haemolymph was 80-100 microg/ml, as determined by porcine IL-18 specific ELISA. This yield was twenty times more than that of the insect cell expression system described previously. The porcine mature IL-18 produced by the silkworms strongly induced interferon gamma (IFN-gamma) production from porcine PBMC. An insect factory system for the large-scale production of useful cytokines for livestock animals will be available in the near future. PMID- 12655118 TI - Mitogen-activated protein kinases partially regulate endothelin-1-induced contractions through a myosin light chain phosphorylation-independent pathway. AB - Endothelin (ET), derived from the endothelium of blood vessels, is a potent vasoactive peptide. Although it has been reported to be involved in cardiovascular diseases, such as hypertension, the mechanism by which ET evokes vasoconstriction is still unclear. On the other hand, p42/p44 mitogen-activated protein kinase (MAPK) and p38 MAPK are activated by a variety of growth factors and cellular stresses, respectively. However, the role of p42/p44 MAPK and p38 MAPK on the ET-1-induced vasoconstriction is not fully understood. This study was undertaken to determine whether p42/p44 MAPK and p38 MAPK participate in the regulation of vascular smooth muscle contraction by ET-1. The isometric vasoconstriction and intracellular Ca(2+) ([Ca(2+)](i)) were simultaneously measured using CAF-100. Phosphorylation of myosin light chain (MLC) and p42/p44 MAPK, p38 MAPK were determined by Western blots. In rat thoracic aorta, ET-1 induced a sustained contraction. In contrast, [Ca(2+)](i) was decreased with time. Both PD98059, an inhibitor of p42/p44 MAPK, and SB203580, an inhibitor of p38 MAPK, partially attenuated ET-1-induced contractions in concentration dependent manners. ET-1 increased phosphorylation of both p42/p44 MAPK and p38 MAPK, and PD98059 and SB203580 completely decreased phosphorylation of p42/p44 MAPK and p38 MAPK in response to ET-1 stimulation, respectively. On the other hand, PD98059 and SB203580 did not affect MLC phosphorylation in response to ET-1 stimulation. These results indicate that p38 MAPK, as well as p42/p44 MAPK, may partially regulate the ET-1-induced contraction through a MLC phosphorylation independent pathway. PMID- 12655119 TI - Development of in situ nest PCR and comparison of five molecular biological diagnostic methods for the detection of intracellular viral DNAs in paraffin sections. AB - Nest polymerase chain reaction (PCR), in situ hybridization (ISH), in situ PCR, in situ PCR/hybridization (PCR-ISH) and in situ nest PCR were compared for the detection and localization of intracellular viral DNAs in paraffin sections. MDBK cells were infected with alcelapine herpesvirus 1 ranging from 10(1) to 10(5) 50% tissue culture infected doses (TCID(50)), incubated 18 hr, then fixed and processed into paraffin blocks. Sections of the cell preparation were subjected to nest PCR, ISH, in situ PCR, PCR-ISH and in situ nest PCR using specific oligonucleotide primers or probes directed against the viral open reading frame 50. In situ nest PCR and nest PCR were found to be capable of detecting the viral DNA in the cells infected with the lowest virus titer. As compared with other molecular biological methods for the detection of the virus, in situ nest PCR was found to be more sensitive than ISH, in situ PCR and PCR-ISH. In situ nest PCR has wide applications for sensitive localization of low copy viral sequences within cells to investigate the role of viruses in a variety of clinical conditions. PMID- 12655120 TI - Involvement of activin and inhibin in the regulation of food and water intake in the rat. AB - The expression of activin and inhibin has been demonstrated in the hypothalamus, but their physiological roles in the brain remain to be elucidated. In the present study, involvement of activin and inhibin in the regulation of food and water intake was examined. Male rats were deprived of food or water for 12 and 60 hr, and mRNA levels of activin/inhibin alpha, betaA and betaB subunits in the hypothalamus were estimated by RT-PCR. Gene expression of alpha subunit transiently decreased at 12 hr of food deprivation, while it did not change during water deprivation. Food and water deprivation for 60 hr increased mRNA levels of betaA and betaB subunits, respectively. These results indicated that gene expression of each subunit was independently regulated. Injection of activin A (0.5 and 4.0 microg) into the third ventricle decreased food intake. Water intake was suppressed by 4.0 microg, but not 0.5 microg, of activin A. Intracerebroventricular injection of inhibin A (0.5 and 4.0 microg) decreased water intake in a dose dependent manner without affecting food intake, suggesting that inhibin could act independently of activin. Taken together, it is suggested that activin and inhibin take part in the central regulation of nutrient and fluid balance, though further study is needed to determine precise molecular species involved. PMID- 12655121 TI - Development of a method of diagnosing ovarian disorders in sows and gilts using uterine ultrasonography. AB - In ultrasonographic diagnosis of ovarian disorders and the estrous cycle in sows, transverse observation of the uterus yielded more characteristic findings than observation of sagittal sections. Transverse ultrasonography revealed that the low progesterone (P) type uterus showed a round structure, while the high P type uterus showed a flattened structure. These results corresponded well with rectal palpation findings: the low P type uterus had a hard, pipe-like structure and the high P type a soft, balloon-like structure. For gilts, we employed a minimum convex type transrectal prostate probe that had an approximately 18 cm insertion handle. The images of the uterus obtained thereby were a similar to those obtained from sows. The above results suggest that it should be possible to diagnose and treat many ovarian disorders in sows and gilts based only on the ultrasonograhic findings. In short, ultrasonograhic findings of a round structure of the uterine wall might be an indication for PMSG (pregnant mare serum gonadotropin) treatment, while findings of a flattened structure might be an indication for PGF(2) (alpha) administration. PMID- 12655122 TI - Inhibition of replication induces non-apoptotic cell death in fibroblast cell lines derived from LEC rats. AB - Hydroxyurea (HU), an anticancer drug, inhibits ribonucleoside diphosphate reductase and reduces pool sizes of deoxyribonucleoside triphosphate (dNTP). The reduction of dNTP results in inhibition of DNA replication. The cytotoxic effect of HU was investigated using fibroblast cell lines from LEC rats. LEC rat cells showed significantly higher sensitivity to HU than did cell lines from control WKAH rats. No significant differences were observed between the percentages of apoptotic cells in either LEC or WKAH rat cells that had been treated with HU and those that had not been treated with HU. LEC rat cells also showed significantly higher sensitivity to aphidicolin, which blocks DNA synthesis by inhibiting DNA polymerase alpha, than did WKAH rat cells. In both LEC and WKAH rat cells, intensified bands of p53 protein were observed immediately after treatment with HU. Although the high level of p53 protein persisted in WKAH rat cells until 6 hr post-incubation time after treatment with HU, the level of p53 protein had decreased at 6 hr post-incubation time in LEC rat cells. When the cells were X irradiated in the absence or presence of HU, the ratio of the surviving fraction without HU to that with HU only slightly increased after X-irradiation in WKAH rat cells. In contrast, the ratio in LEC rat cells significantly increased after X-irradiation in a dose-dependent manner. PMID- 12655123 TI - Development of a mixed antigen agar gel enzyme assay (AGEA) for the detection of antibodies to poxvirus in chicken and turkey sera. AB - A mixed-antigen agar gel enzyme assay (AGEA) was developed to detect antibodies to poxviruses in chicken and turkey sera. The assay combines the principles of immunodiffusion and enzyme assay. For the detection of antibodies to fowl poxvirus (FP), pigeon poxvirus (PP) and turkey poxvirus (TP) in turkey serum samples, the three antigens were combined to form a mixed-antigen assay. To screen for antibodies to FP and PP in chicken serum samples, the two antigens were combined. When FP and PP viruses were combined as antigens, the sensitivity for chicken sera was 64% but the sensitivity of the agar gel precipitation test (AGPT) was 34% (P<0.001). When antibodies were detected in turkey sera using the mixed antigens, the AGEA had a sensitivity of 66.4% while that of AGPT was 25% (P<0.001). PMID- 12655124 TI - Comparison of canine heartworm removal rates using flexible alligator forceps guided by transesophageal echocardiography and fluoroscopy. AB - Fluoroscopy (FS)- or transesophageal echocardiography (TEE)-guided heartworm removal was carried out using flexible alligator forceps to compare the rate of worm removal. As a result, the worm removal rates were similar between the two procedures. However, the TEE-guided procedure does not involve radiation exposure, and facilitates observation of worms in the cardiac chamber and pulmonary artery. Therefore, the TEE-guided procedure is thought to be more useful than the FS-guided procedure in clinical setting. PMID- 12655125 TI - Efficacy of bovine viral diarrhea vaccine used in Japan against bovine viral diarrhea virus type 2 strain 890. AB - Bovine viral diarrhea virus (BVDV) has been segregated into two genotypes, type 1 and type 2. To determine the efficacy of the commercially available bovine viral diarrhea type 1 vaccine used in Japan against BVDV type 2, calves were infected with BVDV type 2 strain 890 4 weeks after administration of the vaccine. The vaccinated calves did not develop any clinical signs and hematological changes such as observed in unvaccinated calves after the challenge. Furthermore, the challenge virus was not recovered from the vaccinated calves throughout the duration of the experiment, whereas it was recovered from all unvaccinated calves. The bovine viral diarrhea vaccine used in Japan is efficacious against infection with BVDV type 2 strain 890. PMID- 12655126 TI - Direct detection of dermatophytes in skin samples based on sequences of the chitin synthase 1 (CHS1) gene. AB - For the direct detection of dermatophytes in skin scrapings and hairs from animals, a primer pair specific to the chitin synthase 1 (CHS1) gene of dermatophytes was constructed. By PCR analysis with the primer pair, dermatophyte DNA could be diagnosed directly and rapidly in clinical skin samples. PMID- 12655127 TI - Peripheral neuroblastoma in a young labrador retriever. AB - A 2-year-old Labrador Retriever developed atrophy of the right temporal muscle, subsequently showed generalized seizure and died 2 months after the clinical onset. Postmortem examination revealed the tumor masses in the right mandibulopharyngeal area, nasopharynx and intracranial space. Histopathologically, these tumor masses were composed of small round neoplastic cells and neuropil-like stroma separated by fibrovascular septa. In the neoplastic masses, small neoplastic cells with round to oval hyperchromatic nuclei and scanty cytoplasm predominated, and angulated neoplastic cells with larger nuclei and moderate cytoplasm were scattered. Immunohistochemically, neoplastic cells were positive for neuron specific enorase, neurofilament protein, chromogranin A, synaptophysin and tyrosine hydroxylase. Based on these findings, this case was diagnosed as peripheral neuroblastoma, presumably originated from the sympathetic ganglion, maybe right craninal cervical ganglion. PMID- 12655128 TI - Molecular cloning of the feline thymus and activation-regulated chemokine cDNA and its expression in lesional skin of cats with eosinophilic plaque. AB - Thymus and activation-regulated chemokine (TARC) is a member of CC chemokine and plays an essential role in recruitment of CC chemokine receptor 4 positive Th2 cells to allergic lesion. To investigate the association of TARC in allergic inflammation of cats, a TARC cDNA was cloned from feline thymus by RT-PCR with 3' rapid amplification of cDNA ends (RACE) method. The feline TARC clone contained a full length open reading frame encoding 99 amino acids which shared 80.8%, 72.5%, 65.6% and 67.8% homology with dog, human, mouse and rat homologues, respectively. Expression of TARC mRNA was detected not only in thymus but also in spleen, lung, lymph node, kidney, small intestine, colon and skin of the normal cat tissues examined. Furthermore, it was found that TARC mRNA was strongly expressed in lesional skin of cats with eosinophilic plaque. The present results demonstrated that TARC might be involved in the pathogenesis of eosinophilic plaque in cats. PMID- 12655129 TI - Changes in the distribution of labeled retinal ganglion cells after an implant of DiI into the optic nerve in the chick embryos. AB - Changes in the distribution of retinal ganglion cells (RGCs) were studied using the retrograde labeling of DiI in chicks and chick embryos. The small retinal area filled with labeled RGCs was observed in the retinal fundus on E8. The labeled retinal area expanded radially toward the peripheral retina as the retina grew, and finally occupied a whole retina by P1. The temporal retina was labeled more rapidly than in the nasal retina. The observed-increasing rate of the labeled area was corrected with the growing rate of the retina. Consequently, the corrected-increasing rate of the labeled area was estimated to be about 390% between E8 and E11, and 20-50% after E11. This means that spreading speed of the maturated RGCs lowered until 1/10-1/20 after E11. PMID- 12655130 TI - Infection of a chimeric simian and human immunodeficiency virus with CCR5 specific HIV-1 envelope to Rhesus macaques. AB - Human immunodeficiency virus (HIV) infects lymphocytes and macrophages via CD4 and chemokine receptors. In this study, the infectivity of a chimeric simian and human immunodeficiency virus (SHIV) having a CCR5-specific HIV-1 envelope gene was examined. A SHIV strain termed SHIV-JRFL could enter cells via CD4 with a chemokine receptor CCR5, not CXCR4, and the viral replication was suppressed by recombinant human RANTES, one of beta-chemokines. The intravenous inoculation of SHIV-JRFL into two rhesus macaques resulted in a systemic infection, though it was rather weak. During the early infection, the production of RANTES from Con A stimulated PBMCs of the infected monkeys increased. These results suggested that beta-chemokine has the potential to limit the infectivity of an R5-type SHIV. PMID- 12655131 TI - Seroprevalence of bovine immunodeficiency virus and bovine leukemia virus in dairy and beef cattle in hokkaido. AB - Serological survey of bovine immunodeficiency virus (BIV) and bovine leukemia virus (BLV) infection was conducted in dairy cattle from 10 different regions of Hokkaido, Japan. Among 390 cattle, 11.0% of cattle were BIV-seropositive and 3.3% were BLV-seropositive. Moreover, in two dairy farms, where bovine leukosis has been reported, prevalence of BIV infections were 6.4 and 9.1%, respectively. In contrast, among 150 beef cattle, 16.6% were BIV-seropositive while none was BLV seropositive. Dual infections with BLV and BIV in dairy cattle were tested by using 107 BLV-seropositive sera, and 20 sera were found BIV-positive (18.7%). These results indicate that BIV infection was widespread in Hokkaido. PMID- 12655132 TI - Association between changes in plasma calcium concentration and plasma tartrate resistant acid phosphatase activity in periparturient cows. AB - Eight periparturient Holstein Friesian cows were examined for plasma tartrate resistant acid phosphatase (TRAP) activity to assess the degree of bone metabolic activity and to evaluate the association between the change in calcium (Ca) concentration and bone metabolism during the periparturient period. Milk fever occurred in 1 of 8 cows just after parturition. Plasma TRAP activities did not markedly change in 5 of 8 cows during the experimental period. The changing rate of Ca between preparturition and just after parturition was under -20% in 3 of 8 cows, and low TRAP activities were observed in 2 of these 3 cows. This study suggests that cows with a low TRAP activity are at risk of developing milk fever in comparison to cows with high TRAP activity. Temporary increases of parathyroid hormone were observed in 7 cows, but not in the cow with milk fever. PMID- 12655133 TI - Leaf-variegated mutations and their responsible genes in Arabidopsis thaliana. AB - Leaf variegation has long been known as a recessive genetic trait in higher plants. Unlike albino mutants, leaf-variegated mutants are non-lethal and thus enable us to study a novel mechanism of plastid development and maintenance. Variegation results from a defect that makes chloroplast development unstable, since at least part of the tissues gives rise to normal chloroplasts. Despite the fact that leaf-variegated mutants have contributed to the findings of maternal inheritance or have been used as genetic markers, these mutations and the responsible loci have been poorly understood at the molecular level. A comprehensive study of the leaf-variegated mutants is possible in Arabidopsis, since such mutants have been known and the cloning can be at relative ease as a model plant. Here I summarize recent progress on characterization of the Arabidopsis leaf-variegated mutants. Detailed analysis of the responsible loci revealed that variegation is caused by a defect in various metabolic pathways related to organelle functions. Thus, studies on these genes provide us with novel redundant mechanisms by which heteroplasmic organelles such as plastids and mitochondria can survive from an environmental stress. PMID- 12655134 TI - DNA polymorphism at the ACAULIS5 locus of the wild plant Arabidopsis thaliana. AB - Nucleotide variation in the ACL5 gene region, which encodes spermine synthase, was analyzed for 21 Arabidopsis thaliana ecotypes and one accession of Arabis gemmifera. In A. thaliana, dimorphism was also detected in the ACL5 region, as in other nuclear genes of this plant. The nucleotide diversity (pi) of the entire region, exon and intron was 0.0163, 0.0042 and 0.0293, respectively. The level of nucleotide variation in this region was among the highest of those reported for genes in this plant species. The neutrality tests of Tajima, and Fu and Li did not detect significant deviation from test assumptions for the polymorphism data. However, the HKA test indicated that the level of polymorphism in the intron was significantly high, compared with A. gemmifera. The high nucleotide variation in the intron is responsible for the high level of nucleotide variation in the entire region. These results can be explained by elevated mutation rate in the ACL5 region in the A. thaliana lineage after the two species were split. PMID- 12655135 TI - Adaptive significance of amylase polymorphism in Drosophila XIII. Old World obscura species subgroup divergence according to biochemical properties of alpha amylase. AB - Biochemical properties of enzyme alpha-amylase were surveyed in Drosophila obscura Old world group of species (D. subobscura, D. ambigua, D. obscura and D. tristis) sampled in the same habitat, with the aim to reveal some ecological and evolutionary aspects of amylase polymorphism, which has been studied extensively in D. subobscura, but not compared with other species in the group. The data obtained show that D. subobscura is distinct from the other three species regarding all biochemical amylase properties. Such a divergence also correlates with the niche breadth and relative abundance of these species in the same habitat. PMID- 12655136 TI - Enhanced cost of mating in female sterile mutants of Drosophila melanogaster. AB - In Drosophila females, mating is known to cause a reduction in life span, which is referred to as 'the cost of mating'. Since mating enhances oogenesis and oviposition, the cost of mating may be regarded as a trade-off between reproduction and longevity. We examined whether the cost of mating exists in mutant females that are unable to produce eggs. Three different mutant alleles of ovarian tumors (otu) and an allele of dunce (dnc(M11)) of Drosophila melanogaster were used to sterilize females. For all the female sterile mutants tested, mating dramatically decreased the life span of homozygous sterile females. Even more extreme shortening of life spans were observed when the sex peptide gene (Acp70A) was expressed in homozygous otu females, though they were virgin, indicating that the shortening in life span is due to seminal factors. These results indicate that the cost of mating is greater in females defective in oogenesis than that in normally fertile females. PMID- 12655137 TI - Phylogenetic relationships in the division Lipastromorphi (Coleoptera, Carabidae) of the world as deduced from mitochondrial ND5 gene sequences. AB - Phylogenetic trees have been constructed using mitochondrial ND5 gene sequences of 85 specimens representing 40 species belonging to the division Lipastromorphi of the subtribe Carabina (= genus Carabus s. lat.) of the world. In the trees constructed by several methods, there have been recognized a basal split into two lineages, A and B, which are further divided into 2 (A1 and A2) and 9 (B1-B9) sublineages, respectively. Each sublineage and the clustering of the species in the trees are more or less linked to their geographic distribution. Morphology of the species within A1 or A2 in the lineage A is similar, and the species in most of the sublineages in the lineage B are common in a number of important morphological characters in spite of their long evolutionary histories. Whilst such a morphological stability, considerable morphological changes may be recognized upon or after split of lineages and sublineages. Based on the molecular phylogeny together with the morphological data, the pattern of diversification is discussed. PMID- 12655139 TI - Phylogeny of the lizard subfamily Lygosominae (Reptilia: Scincidae), with special reference to the origin of the new world taxa. AB - Phylogenetic relationships of the three lygosomine skink genera occurring both in the Old World and the New World (Mabuya, Scincella and Sphenomorphus) were inferred from mitochondrial DNA sequence of 12S and 16S rRNA genes. Results strongly suggested the non-monophyly for any of these three genera. Within the Mabuya group, Asian members appear to have diverged first, leaving the Neotropical and the Afro-Malagasy Mabuya as sister groups. These relationships, together with the absence of extant or fossil representatives of the Mabuya group from North America, strongly suggest the trans-Atlantic dispersals of Mabuya from Africa to Neotropics. Our results also indicated a closer affinity of the New World Scincella with the New World Sphenomorphus than with the Old World Scincella. Such relationships suggest the trans-Beringian dispersal of the common ancestor from Asia and its subsequent divergence into the North American Scincella and the Neotropical Sphenomorphus. PMID- 12655138 TI - Mode of morphological differentiation in the Latitarsi-ground beetles (Coleoptera, Carabidae) of the world inferred from a phylogenetic tree of mitochondrial ND5 gene sequences. AB - The Latitarsi is one large division of the subtribe Carabina (subfamily Carabinae, family Carabidae), and has been considered as a discrete morphological group consisting of 17 genera. The phylogenetic relationships and evolutionary pattern of the Latitarsi ground beetles have been investigated by analyzing mitochondrial NADH dehydrogenase subunit 5 (ND5) gene sequences. The phylogenetic tree suggests that the Latitarsi members do not form a single cluster, i.e., not monophyletic and at least 16 lineages belonging to the so-called Latitarsi emerged at about the same time of the Carabina radiation together with the members of other divisions. This suggests that these lineages (A, B, C, H, L, N, O, P, Q, R, S, T, U, V, W and X in Fig. 2a) may be treated each as a phylogenetically distinct division equivalent to other divisions. The group with bootstrap value of more than 80 percent has been considered as a single lineage (division) with two exceptions, V and X. The independency of each lineage has been assumed by the traditional morphology as well as a single clustering on the trees constructed by independent methods, unchanged topology by replacement of outgroups, etc. Generally speaking, the members in a single lineage are geographically linked. Many phylogenetic lineages are composed of a single or only a few species without conspicuous morphological differentiation. In contrast to such a "silent morphological evolution", a remarkable morphological differentiation occasionally took place in several lineages. PMID- 12655140 TI - Phylogeography and population structure of the Dall's porpoise, Phocoenoides dalli, in Japanese waters revealed by mitochondrial DNA. AB - We investigated genetic diversity and phylogenetic relationships among three morphologically distinct populations of Dall's porpoise (Phocoenoides dalli) in Japanese waters by analyzing mitochondrial DNA variation. These populations, the Sea of Japan-Okhotsk dalli-type population, the truei-type population and the standard dalli-type population in the northwestern North Pacific, are clearly discriminated from each other by differences in the size of their white flank patch. A total of 479 bp of the mitochondrial control region and flanking tRNA genes was sequenced for 103 individuals. Haplotypic diversity was high (h = 0.968), but these haplotypes differed by only a few nucleotides (pi = 0.0106). Although many haplotypes were shared between populations, analysis of molecular variance (AMOVA) indicated genetic subdivision among the three populations (overall F (ST) = 0.023, P < 0.001; phi(ST) = 0.026, P = 0.029). Pairwise comparisons indicated a low but significant difference between the Sea of Japan Okhotsk and the other two populations, whereas there was no significant difference between the latter. These results suggest that there is a close evolutionary relationship among these populations despite their consistent differences in coloration. This may reflect genetic polymorphism in the common ancestral population, which subsequently underwent a rapid divergence. The low genetic variability and haplotypic differentiation of the Sea of Japan-Okhotsk population suggest that it originated from a small population that colonized the Sea of Japan or that experienced population reduction when this Sea was isolated from the North Pacific in the last glacial period. PMID- 12655141 TI - Three novel mutations responsible for Cockayne syndrome group A. AB - Cockayne syndrome (CS) is a rare autosomal recessive disease, which shows diverse clinical symptoms such as photosensitivity, severe mental retardation and developmental defects. CS cells are hypersensitive to killing by UV-irradiation and defective in transcription-coupled repair. Two genetic complementation groups in CS (CS-A and CS-B) have been identified. We analyzed mutations of the CSA gene in 5 CS-A patients and identified 3 types of mutations. Four unrelated CS-A patients (CS2OS, CS2AW, Nps2 and CS2SE) had a deletion including exon 4, suggesting that there is a founder effect on the CSA mutation in Japanese CS-A patients. Patient CS2SE was a compound heterozygote for this deletion and an amino acid substitution at the 106th glutamine to proline (Q106P) in the WD-40 repeat motif of the CSA protein, which resulted in a defective nucleotide excision repair. Patient Mps1 had a large deletion in the upstream region including exon 1 of the CSA gene. Our results indicate that a rapid and reliable diagnosis of CSA mutations could be achieved in CS-A patients by PCR or PCR-RFLP and that the Q106P mutation could alter the propeller structure of the CSA protein which is important for the formation of the CSA protein complex. PMID- 12655142 TI - Construction of a BAC library for Haplochromis chilotes, a cichlid fish from Lake Victoria. AB - Cichlid fishes in Lake Victoria are model organisms for studying rapid radiation and speciation. On the way to examine the molecular basis of how these cichlid fishes achieved such a remarkable morphological diversification, we constructed a bacterial artificial chromosome (BAC) library derived from a cichlid species, Haplochromis chilotes, from Lake Victoria. The library includes 157,056 clones with the average insert size of 128 kb, corresponding to a 10-fold coverage of the H. chilotes genome. Given that the cichlid fishes endemic to Lake Victoria are closely related to one another phylogenetically and their genomes are nearly identical, this BAC library can be utilized to isolate genes from the more than 200 Haplochromine cichlid species in Lake Victoria. PMID- 12655143 TI - The phylogenetic relationships of insectivores with special reference to the lesser hedgehog tenrec as inferred from the complete sequence of their mitochondrial genome. AB - The complete mitochondrial genome of a lesser hedgehog tenrec Echinops telfairi was determined in this study. It is an endemic African insectivore that is found specifically in Madagascar. The tenrec's back is covered with hedgehog-like spines. Unlike other spiny mammals, such as spiny mice, spiny rats, spiny dormice and porcupines, lesser hedgehog tenrecs look amazingly like true hedgehogs (Erinaceidae). However, they are distinguished morphologically from hedgehogs by the absence of a jugal bone. We determined the complete sequence of the mitochondrial genome of a lesser hedgehog tenrec and analyzed the results phylogenetically to determine the relationships between the tenrec and other insectivores (moles, shrews and hedgehogs), as well as the relationships between the tenrec and endemic African mammals, classified as Afrotheria, that have recently been shown by molecular analysis to be close relatives of the tenrec. Our data confirmed the afrotherian status of the tenrec, and no direct relation was recovered between the tenrec and the hedgehog. Comparing our data with those of others, we found that within-species variations in the mitochondrial DNA of lesser hedgehog tenrecs appear to be the largest recognized to date among mammals, apart from orangutans, which might be interesting from the view point of evolutionary history of tenrecs on Madagascar. PMID- 12655144 TI - To reviews on physiological and pathological roles of cell death. PMID- 12655145 TI - The role of Apaf-1 in programmed cell death: from worm to tumor. AB - Apoptosis or programmed cell death is an important process to eliminate unnecessary or hazardous cells. Apaf-1, a mammalian homologue of CED-4 of C. elegans, is the essential adaptor molecule in the mitochondrial pathway of apoptosis. Mice lacking Apaf-1 show accumulation of neurons in the developing central nervous system due to reduced apoptosis. Apaf-1-deficient cells are remarkably resistant to various apoptotic stimuli. Apaf-1-mediated apoptosis plays a role in the prevention of tumorigenesis. However, Apaf-1-independent cell death pathways are also indicated. In this review, we will summarize what has been learned about the role of Apaf-1 by biochemical and genetical approaches. PMID- 12655146 TI - Physiological functions of Pten in mouse tissues. AB - PTEN is a tumor suppressor gene mutated in many human sporadic cancers and in hereditary cancer syndromes such as Cowden disease, Bannayan-Zonana syndrome and Lhermitte-Duclos disease. The major substrate of PTEN is PIP3, a second messenger molecule produced following PI3K activation induced by variety of stimuli. PIP3 activates the serine-threonine kinase PKB/Akt which is involved in anti apoptosis, proliferation and oncogenesis. In mice, heterozygosity for a null mutation of Pten (Pten(+/-) mice) frequently leads to the development of a variety of cancers and autoimmune disease. Homozygosity for the null mutation (Pten (-/-) mice) results in early embryonic lethality, precluding the functional analysis of Pten in various organs. To investigate the physiological functions of Pten in viable mice, various tissue-specific Pten mutations have been generated using the Cre-loxP system. This review will summarize the phenotypes of conditional mutant mice lacking Pten function in specific tissues, and discuss how these phenotypes relate to the physiological roles of Pten in various organ systems. PMID- 12655147 TI - Roles of MAPKKK ASK1 in stress-induced cell death. AB - Apoptosis signal-regulating kinase 1 (ASK1) is a ubiquitously expressed mitogen activated protein (MAP) kinase kinase kinase that activates the c-Jun N-terminal kinase (JNK) and p38 MAP kinase signaling cascades. Recent findings from analyses of ASK1-deficient mice have revealed that ASK1 is required for apoptosis induced by oxidative stress, TNF and endoplasmic reticulum (ER) stress. In addition, several lines of evidence have suggested that ASK1 has diverse functions in the decision of cell fate beyond its pro-apoptotic activity. Thus, ASK1 appears to be a pivotal component not only in stress-induced cell death but also in a broad range of biological activities in order for cells to adapt to or oppose various stresses. PMID- 12655148 TI - Physiological and pathological cell deaths in the reproductive organs. AB - Apoptosis of testicular germ cells and oocytes and their supporting cells in the gonads occurs at physiological and normal conditions or after exposure to pathological stimuli. Cell-death regulators, including Bcl-2 family members, caspases, Fas and p53 are thought to be involved in these processes. This article reviews the details of the apoptotic machinery in the reproductive organs by describing briefly the abnormal phenotypes observed in transgenic and gene ablated mice. PMID- 12655150 TI - The early secretory pathway contributes to autophagy in yeast. AB - Autophagy is a starvation response in eukaryotes by which the cell delivers cytoplasmic components to the vacuole for degradation, and is mediated by a double membrane structure called the autophagosome. We have previously proposed that the specific combination of COPII like components, including Sec24p, is required for autophagy (Ishihara, N. et al. (2001) Mol. Biol. Cell, 12: 3690 3702). The autophagic defect in sec24 deleted mutant cells was, however, suppressed upon the recovery of its secretory flow by the overexpression of its homologue, Sfb2p. We have also reported that the autophagic defect is not observed in sec13 and sec31 mutants, a phenomenon that can be explained by the fact that starvation stress suppresses the secretory defect of these mutants. These observations indicate that the active flow in the early secretory pathway plays an important role in autophagy; that is, autophagy proceeds in the presence, but not in the absence of the early secretory flow. Both autophagy and its closely related cytoplasm to vacuole-targeting (Cvt) pathway occur through a pre-autophagosomal structure (PAS), and since the PAS and the functional Cvt pathway exist in all sec mutants, the early secretory pathway must be involved specifically in autophagy, subsequent to PAS formation. PMID- 12655149 TI - Suppression of cofilin phosphorylation in insulin-stimulated ruffling membrane formation in KB cells. AB - Various cellular events such as cell motility and division are directed by the actin cytoskeleton under the control of its regulatory system. Cofilin is a low molecular weight actin-modulating protein that severs and depolymerizes F-actin and is shown to enhance actin filament dynamics. The activity of cofilin is negatively regulated by phosphorylation at Ser-3. In human epidermoid carcinoma KB cells, insulin treatment induces characteristic ruffling membranes, and it was reported that LIMK1, a cofilin kinase, was activated in these cells treated with insulin. Since cofilin is a key protein responsible for establishing the rapid turnover of actin filaments, it appears to be contradictory that cofilin is phosphorylated (inactivated) by a stimulus that is known to induce the highly dynamic actin structure, ruffling membranes. Therefore, we examined the phosphorylation state of endogenous cofilin in KB cells treated with insulin. The dephosphorylated form of cofilin increased with insulin treatment, as analyzed by nonequilibrium pH gradient gel electrophoresis (NEpHGE)-immunoblotting. Cell labeling with (32)P orthophosphate indicated that cofilin was being continuously phosphorylated and dephosphorylated, and that the apparent insulin-induced dephosphorylation was due to suppression of continuous phosphorylation and not to enhanced dephosphorylation. Further, we examined the localization of the phosphorylated form of cofilin using phospho-specific antibody raised against phosphorylated cofilin. Surprisingly, phosphorylated cofilin was concentrated in the ruffling membranes induced by insulin. These results suggest that the examination of the kinetics and spatial regulation of phosphorylation is critical for the elucidation of the role of cofilin and upstream kinases in actin reorganization. PMID- 12655151 TI - Coculture of endothelial cells and mature adipocytes actively promotes immature preadipocyte development in vitro. AB - Adipose tissue consists of mature adipocytes and endothelial cells, which are all supported by the extracellular matrix. Adipose tissue development is closely associated with angiogenesis. However, the adipocyte-endothelial cell interaction is unclear. To address this issue, we examined the effects of endothelial cells on the growth, apoptosis, and differentiation of mature adipocytes in three dimensional collagen gel culture of the adipocytes with or without rat lung endothelial (RLE) cells. Spindle-shaped preadipocytes, an immature type of adipocyte, developed more actively around the adhesion sites of RLE cells to mature adipocytes in the coculture (rate of preadipocytes: 18.9+/-4.3%) than in the culture of adipocytes alone (2.0+/-5.1%). With respect to growth, RLE cells induced about a three-fold increase in bromodeoxyuridine uptake of mature adipocytes alone, while RLE cells did not influence the uptake of preadipocytes. RLE cells also did not affect the apoptotic indices by immunohistochemistry for single-stranded DNA in mature adipocytes or preadipocytes. These phenomena were not reproduced by RLE cell-conditioned medium, or by certain endothelial cell produced cytokines. Our in vitro study is the first demonstration that endothelial RLE cells promote the active development of preadipocytes together with increased growth of mature adipocytes. These results suggest that endothelial cells are involved in the enlargement mechanism of adipose tissue mass through their direct adhesion to mature adipocytes. PMID- 12655152 TI - Innate apoptosis of human B lymphoblasts transformed by Epstein-Barr virus: modulation by cellular immortalization and senescence. AB - B-lymphoblastoid cell lines (LCLs) transformed by Epstein-Barr virus have a phenotype corresponding to activated B-lymphoblasts. Although they are widely used as models in various biological and medical studies, their innate morphological differentiation and apoptosis has been little studied. We report here that a large proportion of LCL cells spontaneously differentiate into smaller lymphoid cells which ultimately undergo apoptosis during conventional cell culture. Two distinct types of apoptosis with some intermediate types exist: type 1 apoptosis in small and medium-size cells with shrunken nuclei having heavily condensed chromatin in the whole nucleus region accompanied by relatively large internucleosomally fragmented DNA (above 2 kbp); type 2 apoptosis in large lymphoblasts with extremely lobulated nuclei having chromatin condensation beneath the nuclear membrane alone accompanied by smaller internucleosomally fragmented DNA (below 2 kbp). Type 1 apoptotic cells were far more numerous than type 2 apoptotic cells. The incidence of type 1 apoptosis was suppressed by cellular immortalization and was extremely stimulated at the end of the lifespan (crisis). These results provide essential information for us to use LCLs for various biological and medical studies including cellular immortalization, tumorigenesis and senescence. PMID- 12655153 TI - Effects of trypsin-digested outer-arm dynein fragments on the velocity of microtubule sliding in elastase-digested flagellar axonemes. AB - Flagellar movement is caused by the coordinated activity of outer and inner dynein arms, which induces sliding between doublet microtubules. In trypsin treated flagellar axonemes, microtubule sliding induced by ATP is faster in the presence than in the absence of the outer arms. To elucidate the mechanism by which the outer arms regulate microtubule sliding, we studied the effect of trypsin-digested outer-arm fragments on the velocity of microtubule sliding in elastase-treated axonemes of sea urchin sperm flagella. We found that microtubule sliding was significantly slower in elastase-treated axonemes than in trypsin treated axonemes, and that this difference disappeared after the complete removal of the outer arms. After about 95% of the outer arms were removed, however, the velocity of sliding induced by elastase and ATP increased significantly by adding outer arms that had been treated with trypsin in the presence of ATP. The increase in sliding velocity did not occur in the elastase-treated axonemes from which the outer arms had been completely removed. Among the outer arm fragments obtained by trypsin treatment, a polypeptide of about 350 kDa was found to be possibly involved in the regulation of sliding velocity. These results suggest that the velocity of sliding in the axonemes with only inner arms is similar to that in the axonemes with both inner and outer arms, and that the 350 kDa fragment, probably of the alpha heavy chains, increases the sliding activity of the intact outer and inner arms on the doublet microtubules. PMID- 12655154 TI - Development of a conditionally immortalized testicular Sertoli cell line RTS3-3 from adult transgenic rats harboring temperature-sensitive simian virus 40 large T-antigen gene. AB - Transgenic mice and rats harboring temperature-sensitive simian virus 40 (tsSV40) large T-antigen gene are useful for establishing cell lines from tissues. We succeeded in establishing a conditionally immortalized testicular Sertoli cell line, RT3-3, from adult transgenic rats harboring the oncogene. The cells grew at permissive (33 degrees C) and intermediate (37 degrees C) temperatures but not at nonpermissive temperature (39 degrees C). Large T-antigen was expressed at 33 and 37 degrees C, whereas the expression level was gradually decreased at 39 degrees C, suggesting that the temperature-sensitive growth characteristics arise as a result of the function of tsSV40 large T-antigen. The cells showed biochemical features associate with normal Sertoli cells including expressions of mRNAs of sulfated glycoprotein-2 (SGP-2), transferrin (TF) and steel factor. Quantitative polymerase chain reaction revealed that nonpermissive temperature induced increase in the level of SGP-2. Moreover, levels of SGP-2 and/or TF were significantly elevated in the cells treatment with sodium butyrate and retinoic acid, inducers of cellular differentiation. To our knowledge, this is the first report of the establishment of a testicular Sertoli cell line from the transgenic rats. Thus, the conditionally immortalized cell line RTS3-3 with unique characteristics may serve as good experimental in vitro models for basic and applied biology of testicular Sertoli cells. PMID- 12655155 TI - Autoimmune hair loss induced by alloantigen in C57BL/6 mice. AB - Exponentially growing Meth-A cells expressing H-2K(d).D (d) antigen were found to induce alopecia when injected intraperitoneally into normal C57BL/6 mice, which express the H-2K(b).D (b) antigen. However, the capacity to induce alopecia disappeared when Meth-A cells were treated with K252a, which inhibits protein kinases. Histologically, skin in affected areas showed dense mononuclear cell infiltration and a focal foreign-body giant-cell reaction in hair follicles. The subtyping of lymphocytes in peripheral blood demonstrated a significant difference between normal mice and Meth-A cell-injected mice. To further examine the mechanism by which the alloantigen induces alopecia, lymphocytes isolated from the peripheral blood of normal C57BL/6 mice were cultured in medium containing Meth-A cell homogenate, phytohemagglutinin (PHA) and recombinant mouse interleukin-2 (rm IL-2), and intravenously injected into normal C57BL/6 mice. The adoptive transfer of the lymphocytes induced alopecia in a similar way. These findings suggest that the protein kinase-modulated alloantigen induces alopecia by disturbing the immunological homeostasis, and that lymphokine-activated killer cells play an important role in induction of alopecia by cross-reacting with hair follicles. PMID- 12655156 TI - Coronary microembolization. AB - Atherosclerotic plaque rupture is a key event in the pathogenesis of acute coronary syndromes and during coronary interventions. However, it does not always result in complete thrombotic occlusion of the entire epicardial coronary artery with subsequent acute myocardial infarction; in milder forms the result can be embolization of atherosclerotic and thrombotic debris into the coronary microcirculation. This review summarizes the available morphological evidence for coronary microembolization in patients who died from coronary artery disease, most notably from sudden death, and then goes on to address the experimental pathophysiology of coronary microembolization in animal models of acute coronary syndromes and heart failure. Finally, the review presents the available clinical evidence for coronary microembolization in patients, highlights its key features (ie, arrhythmias, contractile dysfunction, infarctlets and reduced coronary reserve) and addresses its prevention by mechanical protection devices and glycoprotein IIb/IIIa antagonism. PMID- 12655157 TI - Sustained reduction of serum cholesterol in low-dose 6-year simvastatin treatment with minimum side effects in 51,321 Japanese hypercholesterolemic patients. AB - The Japan Lipid Intervention Trial (J-LIT) study, a nationwide cohort study utilizing the clinical practice of general physicians, was designed to clarify the relationship between the incidence of coronary heart disease and serum lipid concentrations during simvastatin therapy, as well as the safety of the therapy, in a large number of Japanese hypercholesterolemic patients. All the enrolled patients were treated with simvastatin. The current study analyzed the lipid lowering effect and safety of the low-dose simvastatin therapy used in the J-LIT study. Open-labeled simvastatin was given to 51,321 patients at an initial dose of mostly 5 mg/day. After 6 months of the treatment, the average serum total cholesterol (TC) and low density lipoprotein-cholesterol concentrations in all the patients followed up were reduced by 18.3% and 26.0%, respectively, and that of high density lipoprotein-cholesterol increased 2.3% on average. These concentrations were well maintained throughout the 6-year treatment period. A minority of patients (1.4%) unexpectedly had a remarkable reduction in TC concentration by more than 40%. Hyper-responders, even to low-dose statin, were found for the first time in this large-scale and long-term investigation. Overall adverse drug reactions occurred in 3.3% of subjects during the 6-year treatment, the major events being hepatic and musculoskeletal disorders, of which the incidence was less than 1%. Low-dose simvastatin therapy of 5 mg/day effectively controlled the serum TC concentration by reducing it by approximately 20% on average in hypercholesterolemic Japanese patients, a reduction that corresponds to the effect of simvastatin 20 mg/day in Western studies. In addition, the low incidence of drug-related adverse events in this study may be also related to the low dosage of simvastatin. PMID- 12655158 TI - Intravenous nicorandil in conjunction with coronary reperfusion therapy is associated with better clinical and functional outcomes in patients with acute myocardial infarction. AB - The aim of this retrospective study was to assess whether intravenous nicorandil, a hybrid of NO and a KATP channel opener, in conjunction with percutaneous coronary intervention (PCI) improves the long-term prognosis in patients with acute myocardial infarction (AMI). Intravenous nicorandil has already been shown to improve the in-hospital prognosis of patients with anterior AMI. The study population consisted of 272 patients with a reperfused AMI who were admitted during a similar time interval, before (control; n=114) and after nicorandil treatment (n=158). In the nicorandil group, a 4 mg bolus injection was given, followed by 6 mg/h infusion for 24 h and then oral nicorandil at 15 mg/day for at least 1 month. In the patients with an anterior AMI, left ventricular (LV) function was better and the frequency of LV remodeling was lower after 3 months in the nicorandil group; however, in those with a non-anterior AMI, there were no differences in functional outcome and the frequency of LV remodeling between the 2 groups. The frequency of cardiac events was significantly lower in the nicorandil group, and the use of nicorandil was derived as a potential factor related to freedom from cardiac events (p<0.01, odds ratio = 0.27). Nicorandil treatment was associated with better myocardial perfusion and a better functional and clinical outcome than PCI alone, and this beneficial effect was maintained for a long time, particularly in patients with anterior AMI. PMID- 12655159 TI - Increased serum matrix metalloproteinase-1 concentration predicts advanced left ventricular remodeling in patients with acute myocardial infarction. AB - The left ventricular ejection fraction (LVEF) is one of the major prognostic factors after acute myocardial infarction (AMI) and matrix metalloproteinase-1 (MMP-1) is an enzyme responsible for extracellular collagen degradation and remodeling. The present study investigated whether the concentration of serum MMP 1 was associated with the LVEF after AMI. Blood was sampled on admission, and at 24 h, 3 days, 7 days, 2 weeks and 4 weeks in 24 patients with their first AMI. Left ventriculography was performed 4 weeks after the onset of AMI and the LVEF was calculated by center line method. MMP-1 concentrations were higher at 7 days and at 2 weeks than on admission (p<0.001), and at 7 days (r=-0.655, p=0.0005) and at 2 weeks (r=-0.636, p=0.0008) were negatively correlated with the LVEF. The patients with AMI were divided into high and low LVEF groups according to the results of left ventriculography. Although there were no differences in the clinical characteristics between the 2 LVEF groups, the MMP-1 concentrations at 24 h (p<0.01), 7 days (p<0.01) and 2 weeks (p<0.05) were lower in the high LVEF group than in low LVEF group. A high concentration of MMP-1 at the subacute phase after AMI predicts advanced left ventricular remodeling. PMID- 12655160 TI - Significance of asymmetric dimethylarginine (ADMA) concentrations during coronary circulation in patients with vasospastic angina. AB - The basal activity of nitric oxide (NO) is reduced in spastic arteries of patients with vasospastic angina (VSA). Elevated concentrations of ADMA are associated with reduced NO production and impaired endothelium-dependent vasodilatation. The aim of this study was to elucidate the role of ADMA and its relationship to NO end-products (NOx; nitrate + nitrite) during coronary circulation in patients with VSA. The plasma ADMA and NOx concentrations during coronary circulation were evaluated in 16 VSA and 16 control patients. Blood samples were obtained from the coronary sinus (V) and the ostium of the left coronary artery (A), and the (V-A) differences of ADMA and NOx were determined. The coronary sinus plasma ADMA concentration in patients with VSA was higher than that in the control. The coronary sinus - arterial (V-A) difference of NOx was negative in the VSA group and approximately zero in the control group (VSA group =-1.4 micromol/L, control group =-0.1 micromol/L, p=0.0005). Furthermore, in the VSA patients, there was a negative correlation between the (V-A) difference of NOx and the basal coronary artery tone at the site of spasm (r=-0.60, p=0.015). A significant negative correlation between the (V-A) differences of NOx and ADMA was observed in patients with VSA (r=-0.52, p<0.05), but not in those of the control. Higher ADMA concentrations might cause the reduced formation of NO that underlies the pathophysiology of coronary vasospasm. PMID- 12655161 TI - Risk stratification for asymptomatic patients with Brugada syndrome. AB - Ventricular fibrillation (VF) is induced in some asymptomatic patients with Brugada syndrome (BS), but the prognostic value of programmed electrical stimulation (PES) in such patients is controversial. The clinical characteristics of 41 asymptomatic BS patients, divided into 2 groups according to whether VF was induced by PES (inducible VF group: n=13, non-inducible VF group: n=28) were evaluated. ST levels in the right precordial leads were measured before and after administration of pilsicainide and the abnormal late potential (LP) was evaluated on the signal-averaged electrogram. The ST level at V(2) at baseline in the inducible VF group was significantly higher than that in the non-inducible VF group (p<0.05). Pilsicainide induced significant ST segment elevation in both groups and the ST level after pilsicainide in the inducible VF group was higher than that in the non-inducible VF group (p<0.01). LP was more frequent in the inducible VF group than in the non-inducible VF group. The criterion of ST level >0.15 mV at baseline with pilsicainide-induced additional ST elevation >0.10 mV and positive LP showed high sensitivity (92%) and specificity (89%) for detection of PES-induced VF in asymptomatic BS patients. PMID- 12655162 TI - Sensitive method of detecting myocardial ischemia during dobutamine stress echocardiography. AB - To test the hypothesis that dobutamine-induced myocardial ischemia causes early systolic asynchrony predominantly in the regional left ventricular wall, color kinesis (CK) images during dobutamine stress echocardiography (DSE) were recorded in 13 patients with coronary artery disease and in 10 patients without, all of whom showed normal wall motion at rest. Based on the visual interpretation of DSE and the angiographic findings, 21 segments in the short-axis images at the papillary muscle level were defined as ischemic, and 60 segments of the patients without coronary artery disease were defined as normal. The incremental fractional segmental area change (IFAC) was calculated at 33-ms intervals from the CK images. At the peak dose, IFACs during the first 33 and 33-67 ms were significantly lower in the ischemic segments than in the normal ones, and IFACs during 133-167, 200-233 and 233-267 ms were significantly higher in the ischemic segments. The ratio (peak/low dose) of the cumulative fractional area change at 100 ms gave the best sensitivity (= specificity) for differentiating the 2 groups (86%). Dobutamine-induced ischemia is characterized by an early-systolic asynchrony rather than a change in overall wall excursion and CK can provide an objective assessment of ischemia developing during DSE. PMID- 12655163 TI - Association of body mass index, body fat, and weight gain with inflammation markers among rural residents in Japan. AB - Because obesity is thought to play a key role in atherosclerosis through the low grade chronic inflammation, the present study was designed to investigate associations of body mass index (BMI), body fat, and weight gain with optimized inflammation markers in 1,053 residents who were 40 years of age and older from a rural community (total population = 3,940 in 2000) in Japan. People reporting having a cold and those who did not undergo blood examinations were excluded. C reactive protein (CRP), fibrinogen, serum albumin, and white blood cell (WBC) count were used as the markers for inflammation, body fat was calculated by a conventional method, and weight change since the age of 20 was assessed. The BMI and body fat significantly increased with CRP quartile, and its correlation coefficients to BMI or body fat were relatively high. Similar associations were found for fibrinogen, serum albumin and WBC. Multivariate-adjusted analysis found a high concentration of CRP was significantly associated with obesity, but attenuated the association in other markers. In an analysis restricted to people aged 40-69 years, body fat levels were more strongly associated with CRP and fibrinogen than with BMI only. Furthermore, only CRP concentrations were significantly elevated according to weight gain. Strong associations of CRP concentration with BMI, body fat, and weight gain were found among elderly Japanese, but not with fibrinogen, serum albumin or WBC. PMID- 12655164 TI - Clinical features of acute pulmonary thromboembolism in younger patients. AB - The incidence of acute pulmonary thromboembolism (APTE) in younger patients is extremely low compared with older patients, so the clinical features of these younger patients with APTE is unknown. In the present study, 8 patients with APTE who were less than 40 years old (YG) were compared with 40 patients who were more than 41 years of age (OG). All YG patients had coagulopathy compared with 3 patients in the OG (p<0.01). Deep venous thrombosis (DVT) occurred in all 8 patients in the YG compared with 19 patients in the OG (p<0.01). A higher incidence of patients whose symptoms occurred gradually was noted in the YG (p<0.05). There were no significant differences in clinical characteristics, initial symptoms, past history or other predisposing factors for venous thromboembolism between the 2 groups. Residual pulmonary hypertension was not noted in the YG. However, 1 patient in the YG had recurrent APTE, despite good warfarin control. This study demonstrated the frequency of gradual onset, coagulopathy and clinical signs of DVT in the YG and therefore more careful and long-term observation is necessary in such patients. PMID- 12655165 TI - Protective effect of basic fibroblast growth factor against myocyte death and arrhythmias in acute myocardial infarction in rats. AB - The present study in rats investigated whether basic fibroblast growth factor (bFGF) plays an important role in cardioprotection against myocardial cell death and arrhythmias in acute myocardial infarction (AMI). After ligating the left coronary artery in 62 Wistar rats, 20 Eg of human recombinant bFGF was injected into the infarcted myocardium in 33 rats (group F), while saline was used for 29 control rats (group C). The development of ventricular tachyarrhythmias was assessed during the first 30 min of ischemia. After 24 h occlusion, the hearts of the surviving rats (group F: n=13, group C: n=10) were excised to assess minimum infarct wall thickness and infarct size, determine the number of TUNEL-positive cardiomyocytes and to analyze Bcl-2 and Bax expression by immunohistochemical staining and Western blotting. The incidence of ventricular tachycardia was higher in group C than in group F (p<0.05). The thinning ratio was higher in group F than in group C (p<0.05). There were fewer TUNEL-positive cardiomyocytes in the infarct border area in group F than in group C (p<.0001). Western blot analysis showed greater expression of Bcl-2 in group F than in group C (p<0.05), but similar expression of Bax in the 2 groups. In conclusion, intramyocardial administration of bFGF prevented ischemia-induced myocardial cell death and arrhythmias. PMID- 12655166 TI - Effect of pilsicainide on atrial electrophysiologic properties in the canine rapid atrial stimulation model. AB - The heterogeneous process of atrial electrical remodeling (AER) in the canine rapid atrial stimulation model has been previously reported although it has been reported that a sodium channel blocker might suppress the shortening of the atrial effective refractory period (AERP), its effect on long-term electrical remodeling is unknown. In the present study, the effect of pilsicainide on AER was evaluated. The right atrial appendage (RAA) was paced at 400 beats/min for 2 weeks. In the RAA, Bachmann's bundle (BB), the right atrium near the inferior vena cava (IVC) and in the left atrium (LA), AERP, AERP dispersion (AERPd) and the inducibility of atrial fibrillation (AF) were evaluated at several time points of the pacing phase and the recovery phase (1 week). The same protocol was performed during the administration of pilsicainide (4.5 mg/kg per day) and the parameters were compared with the controls. In the control dogs, the AERP was significantly shortened by rapid pacing at all atrial sites studied and the AERP shortening (DeltaAERP) was larger at the RAA and LA sites (p<0.03). However, pilsicainide decreased these DeltaAERPs at all 4 atrial sites. AERPd was increased during the pacing phase whereas it was decreased during the recovery phase in the control dogs. In contrast, this pacing-induced AERPd was attenuated by the administration of pilsicainide. The AF inducibility was highest at the LA site in both groups, and the inducibility was lower in the pilsicainide group than the control group at all atrial sites. During the rapid pacing phase, the ventricular heart rate was significantly lower in the pilsicainide group than the control because of intra-atrial conduction block. In a canine rapid right atrial stimulation model, pilsicainide suppressed the shortening of the AERP at all atrial sites, possibly through the improvement of the hemodynamics as well as the action of the Na - Ca exchanger. PMID- 12655167 TI - TNF-alpha rapidly antagonizes the beta-adrenergic responses of the chloride current in guinea-pig ventricular myocytes. AB - The purpose of this study was to test the hypothesis that tumor necrosis factor alpha (TNF-alpha) rapidly antagonizes the beta-adrenergic responses of the chloride current and to clarify the intracellular mechanisms responsible for the anti-adrenergic action. The whole-cell patch-clamp technique was used to monitor the anti-adrenergic effects of TNF-alpha on the cAMP-dependent chloride current (I(Cl)) recorded from isolated guinea-pig ventricular myocytes. Ramp pulses (+/ 120 mV; dv/dt = +/-0.4 V/s) were applied from the holding potential of -40 mV. TNF-alpha rapidly (<15 min) inhibited the isoproterenol (Iso, 0.1 micromol/L) induced I(Cl) in a concentration-dependent manner (30-1,000 U/ml, IC (50) = 144 U/ml, n=30). The inhibitory action of TNF-alpha was also observed when I(Cl) had been previously stimulated by 1 micromol/L forskolin (n=5). Prior exposure of myocytes to 5 microg/ml pertussis toxin (PTX) hardly affected the anti-adrenergic action of TNF-alpha (n=4). However, when I(Cl) was induced by both 8-bromo-cAMP (100 micromol/L) and isobutylmethylxanthine (0.1 mmol/L), TNF-alpha (1,000 U/ml) failed to decrease I(Cl) amplitude (n=5). Prior exposure of myocytes to 5 mg/ml pertussis toxin (PTX) hardly affected the anti-adrenergic action of TNF-alpha (n=4). Furthermore, despite of the presence of nitro-L-arginine methyl ester (0.1 mmol/L), a nitric oxide synthase (NOS) inhibitor, TNF-alpha reversed the Iso induced increase in I(Cl) (n=5). These results suggest that TNF-alpha rapidly antagonizes the beta-adrenergic responses of I(Cl) by reducing cAMP concentration. This anti-adrenergic action is mediated by neither the PTX sensitive G proteins regulatory pathway nor constitutive NOS activation. PMID- 12655168 TI - Congenitally corrected transposition of the great arteries treated by partial systemic ventriculectomy. AB - Congenitally corrected transposition of the great arteries (CCTGA) is a rare congenital heart disease characterized by atrioventricular (AV) and ventriculo arterial discordance;(1) that is, the left ventricle supports the pulmonary circulation and the right ventricle supports the systemic circulation. The most common cardiac anomalies in CCTGA include ventricular septal defect, pulmonary outflow tract obstruction and abnormalities of the systemic AV valve. (1) The dysfunction of the systemic ventricle occurs with increasing frequency in older patients with CCTGA, independent of their commonly associated structural defects.(2-4) We report a patient with CCTGA undergone the partial ventriculectomy for his systemic ventricular dysfunction. PMID- 12655169 TI - Mallory - weiss tear complicating intraoperative transesophageal echocardiography. AB - A Mallory - Weiss tear occurred as a complication of intraoperative transesophageal echocardiography carried out in a 62-year-old man who underwent coronary artery bypass grafting. Left ventricular function was monitored in the transgastric short-axis view. Postoperative esophagogastroscopy revealed a Mallory - Weiss tear at the gastroesophageal junction and erosions in the cardia, presumably secondary to contact pressure by the echoprobe and ultrasonic thermal injury. When not actively imaging, the echoprobe should be left free in the esophagus with the acoustic power off. PMID- 12655170 TI - Chronic traumatic pseudoaneurysm of the ascending aorta causing right ventricular inflow obstruction. AB - A 28-year-old man developed exertional dyspnea and peripheral edema 1 year after the initial blunt trauma to the chest. The diagnosis of pseudoaneurysm (PSA) of the ascending aorta was made based on the echocardiographic finding of a large echo-free space that communicated with the ascending aorta through the defect in the ascending aortic wall. The PSA caused a right ventricular inflow obstruction with increased diastolic transtricuspid blood flow velocity up to 2.7 m/s. Following the excision of the PSA and repair of the communication between it and the ascending aorta, the sizes of the right atrium and right ventricle, and the diastolic transtricuspid blood flow velocity, normalized. PMID- 12655171 TI - Successful catheter interventional therapy for acute coronary syndrome secondary to kawasaki disease in young adults. AB - Acute coronary syndrome occurred in 2 young adults who had a history of Kawasaki disease (KD), but few other coronary risk factors. The first patient was a 27 year-old male with acute myocardial infarction without stenosis detected by coronary arteriography 4 years earlier. Emergency coronary arteriography showed occlusion of the right coronary artery. Aspiration-thrombectomy and rescue balloon angioplasty were successfully performed. The second patient was a 32-year old male with unstable angina. Right coronary arteriography showed total occlusion with severe calcification. Left coronary arteriography showed 99% stenosis at the proximal site of the circumflex artery, and a directional coronary atherectomy was performed. Histological examination of a specimen from this site revealed a lipid core, macrophages, and smooth muscle cells. Restenosis was not observed on follow-up coronary arteriography after 5-6 months in either case. The coronary stenosis in each case was probably caused by accelerated atherosclerosis at the site without aneurysm as it seemed to be 'normal' on arteriography. Conventional catheter intervention was effective treatment. The sequelae of KD should be recognized as independent coronary risk factors. PMID- 12655172 TI - Disappearance of intima over the stent and ulcer formation after intracoronary radiation for in-stent restenosis. AB - Little is known about the alterations of the vascular surface after radiation therapy for in-stent restenosis in humans, even though animal experiments suggest that delayed healing of the neointima is a cause of late thrombosis. Coronary angioscopy, together with coronary angiography, was performed at 3 months follow up of 7 patients with in-stent restenosis who underwent beta-radiation therapy. Minimal lesion diameter (MLD) of the lesion increased from 1.00+/-0.30 mm (immediately before) to 2.44+/-0.39 mm (immediately after) and the MLD was well maintained 3 months later (2.34+/-0.62 mm) without any cases of restenosis. In 5 patients, the intima was so thin that some stent struts could be seen through it on angioscopy and in 2 of those, the intima over the stent had disappeared and 1 patient showed ulceration of the vascular wall beneath the stent. After intracoronary radiation therapy, the intima can become so thin that some stent struts are exposed to the lumen, which may be related to the occurrence of late thrombosis. Accordingly, patients who are treated with intracoronary radiation therapy may need long-term antiplatelet therapy. PMID- 12655174 TI - Effect of acute L-tryptophan exposure on the brain serotonergic system and behavior in the male medaka. AB - The aim of this study was to investigate the effect of exposure to L-tryptophan (TRP) on the metabolism of 5-hydroxytryptamine (5HT) and behavior of medaka. In the first experiment, the fish were exposed to a 0, 1, 2 or 4 g/l of TRP solution for 24 hr. Although no significant difference in the brain 5HT content was detected, 5-hydroxyindoleacetic acid (5HIAA), a major 5HT metabolite, increased in a dose-dependent fashion. In the second experiment, the fish were maintained in a 0 or 4 g/l of TRP solution for 28 hr, and then their behaviors were monitored. The fish reared in under TRP solution were divided into two groups and transferred to either fresh water or a TRP solution. The locomotion of the TRP treated group significantly increased compared to the control group irrespective of water conditions. It was suggested that TRP exposure activated the brain 5HTnergic systems and stimulated behavior of medaka. PMID- 12655173 TI - Effects of 20-hydroxyecdysone and serotonin on neurite growth and survival rate of antennal lobe neurons in pupal stage of the silk moth Bombyx mori in vitro. AB - Effects of 20-hydroxyecdysone and serotonin on the morphological development and the survival of antennal lobe neurons from day-2 pupal brains of the silk moth Bombyx mori were investigated in vitro. Four morphologically distinct neuronal types could be identified in the cultured antennal lobe neurons: unipolar, bipolar, multi-polar and projection neurons. Antennal lobe neurons in culture with 20-hydroxyecdysone and serotonin showed different patterns of the morphological development from those described in Manduca sexta. Projection neurons extend their neurites remarkably by 20-hydroxyecdysone in B. mori, but there is no extension from antennal lobe neurons in M. sexta. Multi-polar neurons conspicuously increase only formation of new branches from their primary neurites by serotonin in B. mori, but there are both extension and branching of the neurites in M. sexta. On day-5, antennal lobe neurons in lower titers of 20 hydroxyecdysone had significantly higher survival rates than those in higher titers. Neurons cultured for 7 days at different levels of 20-hydroxyecdysone generally showed significantly lower survival rates than neurons cultured for 5 days under the same conditions. PMID- 12655175 TI - Design of the labial cuticle in Cenocorixa bifida Hung. (Hemiptera: Corixidae) with reference to ionic transport. AB - The surface topography and ultrastructure of the labial cuticle of Cenocorixa bifida were examined by scanning and transmission electron microscopy. The dorsal wall of the labium consists of seven sclerotized transverse bars each displaying two rows of semicircular grooves and pores. The cuticle is about 20 microm thick and is composed of epicuticle and lamellate exocuticle and endocuticle, the latter separated from the underlying epidermis by subcuticle containing amorphous material. The epicuticle is subdivided into an electron-dense very thin outer epicuticle and a homogenous thick inner epicuticle, which is penetrated by grooves. The exocuticle is filled with electron-dense blocks of material, which may provide mechanical support to the labial wall. The elongate epidermal cells display extensive infoldings of the apical plasma membrane (facing the cuticle) and contain abundant mitochondria in the cytoplasm. The presence of deep epicuticular grooves and pores in the thin labial cuticle and extensive apical membrane infolding and abundant mitochondria in the epidermal cells suggest that the labium in C. bifida is the site of osmoregulatory ionic uptake. PMID- 12655176 TI - Expression of membrane-bound and soluble guanylyl cyclase mRNAs in embryonic and adult retina of the medaka fish Oryzias latipes. AB - Localization of mRNAs for four membrane-bound guanylyl cyclases (membrane GCs; OlGC3, OlGC4, OlGC5, and OlGC-R2), three soluble guanylyl cyclase subunits (soluble GC; OlGCS-alpha(1), OlGCS-alpha(2), and OlGCS-beta(1)), neuronal nitric oxide synthase (nNOS), and cGMP-dependent protein kinase I (cGK I) was examined in the embryonic and adult retinas of the medaka fish Oryzias latipes by in situ hybridization. All of the membrane GC mRNAs were detected in the photoreceptor cells of the adult and embryonic retinas, but in different parts; the OlGC3 and OlGC5 mRNAs were expressed in the proximal part and the OlGC4 and OlGC-R2 mRNAs were expressed in the outer nuclear layer. The mRNA for nNOS was expressed in a scattered fashion on the inner side of the inner nuclear layer in the adult and embryonic retinas. The mRNAs (OlGCS-alpha(2) and OlGCS- beta(1)) of two soluble GC subunits (alpha(2) and beta(1)) were expressed mainly in the inner nuclear layer and the ganglion cell layer of the embryonic retina while the mRNAs of the soluble GC alpha(1) subunit and cGK I were not detected in either the adult or embryonic retina. These results suggest that NO itself and/or the cGMP generated by soluble GC (alpha(2)/beta(1) heterodimer) play a novel role in the neuronal signaling and neuronal development in the medaka fish embryonic retina in addition to the role played by phototransduction through membrane GCs in the adult and embryonic retinas. PMID- 12655178 TI - Regulation of Wolbachia density in the Mediterranean flour moth, Ephestia kuehniella, and the almond moth, Cadra cautella. AB - The Mediterranean flour moth, Ephestia kuehniella, is infected with A-group Wolbachia (wKue), and the almond moth, Cadra cautella, is doubly infected with A- and B-group Wolbachia, which are designated as wCauA and wCauB, respectively. In both insects, the Wolbachia populations increased greatly during embryonic and larval stages. The Wolbachia population doubled every 3.6 days on average in E. kuehniella larvae, whereas those of wCauA and wCauB doubled every 2.1 days in C. cautella larvae. The populations of wCauA and wCauB that had been transferred into the E. kuehniella background increased at similar rates to that of wKue in the natural host E. kuehniella, suggesting that the host genetic background influences Wolbachia proliferation. To examine whether the populations of the two Wolbachia variants in double infection is regulated collectively or independently, we measured the infection load in the ovaries of three transfected E. kuehniella lines in different infection states: single infection with wCauA, single infection with wCauB, and double infection. The density of each Wolbachia variant did not differ significantly between the singly and doubly transfected hosts, suggesting independent regulation. PMID- 12655177 TI - A novel G protein alpha subunit in embryo of the ascidian, Halocynthia roretzi. AB - A cDNA clone encoding a novel G protein alpha subunit, HrGalpha(n) was isolated from the larvae of ascidian, Halocynthia roretzi. In contrast with overall amino acid identity (63%) with G protein alpha subunit of G(i) or G(o) subclass, HrGalpha(n) has a unique amino acid sequence, which lacks a residue for pertussis toxin substrate, but retains for cholera toxin substrate for ADP-ribosylation. The sequence characteristics and molecular phylogenetic analysis suggest that HrGalpha(n) defines a novel subclass within G(i) class of G protein alpha subunits. The zygotic expression of HrGalpha(n) was first detected at the 64-cell stage and observed in all blastomeres except for B7.4, B7.5 and B7.6 cells till the 110-cell stage. As progress of the developmental stages, the expression of HrGalpha(n) became restricted and was observed in the muscle, mesenchyme and a part of trunk lateral cells in tailbud embryos. With HrGalpha(n)-GFP fusion-gene construct it was showed that the genomic fragment containing 2674 bp upstream of the putative translation start site of HrGalpha(n) contained the regulatory sequence responsible for the expression in the muscle and mesenchyme cells, and that the regulatory sequence functioned also in Ciona intestinalis. Our results suggest a possible involvement of HrGalpha(n) in the signaling system regulates the cell fate during the embryogenesis of the ascidian. PMID- 12655179 TI - Oryzias curvinotus has DMY, a gene that is required for male development in the medaka, O. latipes. AB - DMY is a Y-specific DM-domain gene required for male development and appears to be the sex-determining gene in the teleost fish medaka, Oryzias latipes. Although the genomic region containing DMY appears to have originated through duplication of the DMRT1 region, it is unknown when the duplication occurred. Here we show that O. curvinotus also has the DMY gene on the Y chromosome, which is homologous to the Y chromosome of medaka, and that DMY is expressed in XY embryos. A phylogenetic tree based on the amino acid sequence including the DM-domain shows that DMY was derived from DMRT1 immediately before speciation of O. latipes and O. curvinotus. PMID- 12655180 TI - Atlas of the embryonic brain in the pygmy squid, Idiosepius paradoxus. AB - Gross structural changes and neuropil formation in the brain during development were described in Idiosepius paradoxus, a sepioid that we chose as a model cephalopod. The brain originates in 4 pairs of ectodermal placodes, which occur separately in the embryonic surface undergoing epiboly. In the final period of epiboly, neuroblasts internalize from the placodes and gather into 4 pairs of ganglionic masses. The ganglionic masses assemble into a ring-like cluster encircling the inner yolk and the foregut anlage, gradually integrated into the 4 domains of a massive brain, a subesophageal mass (SBM), a supraesophageal mass (SPM), and a pair of optic lobes. In the early brain, neuropil forms a framework composed of a longitudinal ladder lying in the SBM, and a transverse arch standing on the lateral sides of the SBM and crossing the SPM. Differentiation of brain lobes proceeds from ventral to dorsal along this framework; first the magnocellular lobes and the posterior pedal lobe appear first in the SBM, the other lobes in the SBM and the basal lobes follow in the proximal region of the SPM, and the accessory lobes develop last in the most dorsal zone of the SPM. In the hatchlings, the brain lobes show almost the same arrangement as in the adults, but the accessory lobes, particularly the vertical lobe, are much smaller than those in the adults. Comparison of the present results with those in the teuthoid and the octopod indicates that developmental sequences of the brain are highly conserved in the coleoid cephalopods. PMID- 12655181 TI - Localization of the nitric oxide/cGMP signaling pathway-related genes and influences of morpholino knock-down of soluble guanylyl cyclase on medaka fish embryogenesis. AB - To better understand the nitric oxide (NO) / cyclic GMP (cGMP) signaling pathway during embryogenesis, we examined the spatial and temporal expression pattern of the genes for neuronal nitric oxide synthase (nNOS), soluble guanylyl cyclase (soluble GC) subunit (OlGCS-alpha(1), OlGCS-alpha(2), and OlGCS-beta(1)), and cGMP-dependent protein kinase (cGK) I and II (cGK I and cGK II) in the medaka fish embryos. OlGCS-beta(1) and nNOS were expressed maternally and OlGCS alpha(1), OlGCS-alpha(2),and cGK II were expressed zygotically. The zygotic expression of OlGCS-alpha(1) and cGK I was detected at stage 19, while that of OlGCS-alpha(2) was detected at stage 16. Whole-mount in situ hybridization showed that the expression of nNOS or cGK I was localized in tail bud, otic vesicles, thyroid, and brain ventricle, or in thymus, gill arch, and olfactory pits, respectively, and that of OlGCS-alpha(1), OlGCS-alpha(2), or OlGCS-beta(1) was dim and dispersed throughout the embryos. To clarify the "role of the NO/cGMP signaling pathway in embryogenesis, we examined the influences of morpholino antisense oligonucleotide of the soluble GC subunit gene (alpha(1)-MO, alpha(2) MO or beta(1)-MO) on development of medaka fish embryos. Embryos injected with alpha(1)-MO or alpha(2)-MO mainly exhibited abnormalities in the central nervous system, including defects in the formation of forebrain, eye, and otic vesicles. alpha(2)-MO injection caused cell death at the tail bud of the embryos at stage 22, and beta(1)-MO injection inhibited the development of the embryos at late blastula. These results suggest that the NO/cGMP signaling pathway plays critical roles in early embryogenesis. PMID- 12655182 TI - Correlation of mitogen-activated protein kinase activities with cell survival and apoptosis in porcine granulosa cells. AB - The regulation of granulosa cell survival and death is critical for determining the fate of ovarian follicles. Mitogen-activated protein kinases (MAPKs) play central roles in various cellular responses, but the relationship between MAPK activities and granulosa cell survival as well as death is poorly understood. The present study examines the roles of the extracellular signal-regulated kinase (ERK) and p38 MAPK activities in porcine granulosa cells in response to survival factors and oxidative stress. Cell survival and apoptosis were evaluated by Trypan blue staining, DNA fragmentation, and chromatin staining with Hoechst 33342. Cell survival induced by serum or by follicle-stimulating hormone (FSH) was inhibited when ERK activity was attenuated with PD98059, which led to the induction of apoptosis. The p38 inhibitor SB203580 significantly decreased the cell survival evoked by FSH, but not by serum. Even in the presence of 10% serum, H(2)O(2) caused apoptosis, indicating that H(2)O (2) may be an atretogenic factor or its mediator. Interestingly, this induction of apoptosis was also prevented by SB203580, suggesting that p38 is involved in an apoptotic pathway induced by H(2)O (2) as well as in a survival pathway evoked by FSH in granulosa cells. These results indicate that whereas ERK activity is critical to the survival of granulosa cells, p38 activity contributes to their survival or apoptosis depending on the stimulus. PMID- 12655183 TI - Cloning of the gene for the thyrotropin beta subunit in the Japanese crested ibis, Nipponia nippon. AB - We isolated a putative gene for the thyrotropin beta subunit (TSHbeta) from two types of genomic libraries of the Japanese crested ibis, Nipponia nippon. Exon intron structure was deduced by comparing the determined sequence with those of TSH beta cDNA of other birds. The deduced amino acid sequence shows extensive similarities to those of the other birds, which assures our assumption that the acquired nucleotide sequence represents the TSHbeta gene. The assembled genomic fragment is 4192 bp in size and consists of 1937 bp of putative 5' flanking region followed by exon-intron structure with three exons and two introns, similar to those observed in rat, human and goldfish counterparts. Locations of introns are also similar to those in mammals and goldfish. Comparison of the 5' flanking region of the ibis TSHbeta gene with those of mammals reveals that several regulatory sequences, such as negative thyroid hormone responsive element (nTRE), Pit-1 responsive element, and AP-1 responsive element, which were characterized in mammalian TSHbeta genes, are also found in the promoter region. This is the first report on the exon-intron structure and 5' flanking region of the TSHbeta gene in an avian species. PMID- 12655185 TI - A new species of parastenhelia (Copepoda: Harpacticoida: Parastenheliidae) from Korea. AB - A new species belonging to the genus Parastenhelia of family Parastenheliidae is described on the basis of the specimens of both sexes collected from the zosteran bed at Seungbong Island and from the shallow sandy bottoms of Jeju Island in Korea. It is distinguished from the congeners by its character combination of inserted location and the length proportion of the inner seta on the first endopodal segment of leg 1, shape of female leg 5, the modified male legs 2-3 endopods, and the number of setae on male leg 5 exopod. An emended table of morphological characters for the genus is provided. PMID- 12655184 TI - Steroidogenic acute regulatory protein in eels: cDNA cloning and effects of ACTH and seawater transfer on its mRNA expression. AB - Steroidogenic acute regulatory protein (StAR) is a key molecule for steroid production by translocating cholesterol from the outer to inner mitochondrial membrane. Two cDNAs of different length encoding StAR was cloned from the head kidney of the eel (Anguilla japonica). In the 3'-untranslated region (UTR) of the longer cDNA, two putative polyadenylation signals were found. The shorter one differed from the longer one solely by the lack of middle of 3'-UTR including the first polyadenylation signal. Reverse transcription-polymerase chain reaction (RT PCR) that differentiates the two mRNAs showed that the ratio of the two was highly variable among individuals, and no preferential expression was detected between freshwater and seawater eels. The predicted protein consists of 285 amino acid residues with 64-83% identity to other StARs thus far obtained. RT-PCR analyses revealed that eel StAR mRNA was expressed abundantly in the head kidney and gonad, and faintly in the brain; but no expression was detected in the gill, heart, liver, intestine, kidney and skeletal muscle. Plasma cortisol concentration increased, but StAR mRNA content in the head kidney did not change, 3 and 24 h after transfer of freshwater eels to seawater, indicating that the transcriptional regulation of StAR may not be involved in cortisol production after seawater transfer. However, ACTH elevated both plasma cortisol and StAR mRNA levels in the head kidney 1.5 and 4.5 h after injection. Thus, the steroidogenic effect of ACTH is mediated by increased StAR production as observed in mammals. PMID- 12655186 TI - Two new freshwater species of the genus Jesogammarus (Crustacea: Amphipoda: Anisogammaridae) from northern Japan. AB - Two new anisogammarid species belonging to the genus Jesogammarus were obtained from freshwater habitats from northern Japan. They are Jesogammarus (J.) fujinoi sp. nov. and Jesogammarus (J.) shonaiensis sp. nov. The former new species is diagnosed by the pleonites without spine and the telson without distolateral spine. The other new species differs from the congeneric species in having many dorsal spines on pleonites 1-3. PMID- 12655187 TI - Phylogenetic relationships and divergence times among mustelids (Mammalia: Carnivora) based on nucleotide sequences of the nuclear interphotoreceptor retinoid binding protein and mitochondrial cytochrome b genes. AB - Phylogenetic relationships among 20 species-group taxa of Mustelidae, representing Mustelinae (Mustela, Martes, Gulo), Lutrinae (Enhydra), and Melinae (Meles), were examined using nucleotide sequences of the nuclear interphotoreceptor retinoid binding protein (IRBP) and mitochondrial cytochrome b genes. Neighbor-joining and maximum-parsimony phylogenetic analyses on these genes separately and combined were conducted. While IRBP performed better than cytochrome b in recovering more-inclusive clades, cytochrome b demonstrated more resolving power in recovering less-inclusive clades. Strong support was found for a close affinity of Enhydra with Mustela to the exclusion of Martes and Gulo (causing Mustelinae to be paraphyletic); the most-basal position of Mustela vison within Mustela, followed by Mustela erminea; an association of Mustela lutreola, Mustela itatsi, Mustela sibirica, and the subgenus Putorius (including Mustela putorius and Mustela eversmanii), to the exclusion of Mustela nivalis and Mustela altaica; and a basal position of Mustela itatsi to a clade containing Mustela sibirica and Putorius. Whereas cytochrome b strongly supported Mustela lutreola as the sister species to Putorius, IRBP strongly supported its basal placement to the Mustela itatsi-Mustela sibirica-Putorius clade. The low level of sequence divergence in cytochrome b between Mustela lutreola and Putorius is therefore a result of interspecific mitochondrial introgression between these taxa, rather than a recent origin of Mustela lutreola in a close relationship to Putorius. Time estimates inferred from IRBP and cytochrome b for mustelid divergence events are mostly in agreement with the fossil record. PMID- 12655189 TI - [Our clinical experience with non-cemented total hip revision arthroplasty]. AB - OBJECTIVES: We evaluated the results of non-cemented revision arthroplasty in patients who had undergone cemented total hip arthroplasty. METHODS: Twenty patients (12 women, 8 men; mean age 72 years; range 61 to 84 years) underwent non cemented revision arthroplasty after a mean of 9.4 years (range 3 to 16 years) following primary surgery. The mean follow-up period was 4.5 years (range 3 months to 6.3 years). RESULTS: During revision arthroplasty, femur fractures were observed in three patients (15%). Three patients (15%) had femoral cortex perforations. Trochanteric osteotomy was performed in five patients (25%). The mean Harris hip score after the operation was 76.7 (range 50 to 90). The results were excellent in three patients (15%), good in seven (35%), fair in seven (35%), and poor in three patients (15%). CONCLUSION: Despite a high intraoperative complication rate during non-cemented revision total hip arthroplasty, we believe that non-cemented revision should be the preferred method because of low infection and low aseptic loosening rates. PMID- 12655188 TI - Genetic relationships and origin of two geographic groups of the freshwater threespine stickleback, 'hariyo'. AB - 'Hariyo' comprise the southernmost freshwater populations of the threespine stickleback, Gasterosteus aculeatus species complex, distributed in central Honshu, Japan. Two geographic groups (Gifu and Shiga) of the 'hariyo' have been recognized as differing from each other in some morphological and ecological features. In order to elucidate the genetic characteristics and phylogenetic position of these populations, partial sequences of the mitochondrial cytochrome b gene were compared in 123 specimens from 10 'hariyo' populations, and eight other freshwater and anadromous populations of threespine stickleback in Japan. Phylogenetic analysis resolved 22 haplotypes in a single most-parsimonious tree. In contrast to previous allozyme analyses, the haplotype tree indicated that 'hariyo' populations are monophyletic. Each of two geographic groups of the 'hariyo' was also shown to be nearly monophyletic. The two geographic groups differed from each other in sequence by an average of 0.47% and from other populations by 0.70%. Based on a molecular clock, constructed from fossil records and mtDNA genetic distances of Gasterosteus and Pungitius, it was estimated that the two groups of 'hariyo' differentiated from the other threespine stickleback populations and also from each other in the middle Pleistocene. The results suggested that each of the two groups of 'hariyo' is a distinct evolutionarily significant unit with unique genetic features, as well as morphological and ecological characteristics. PMID- 12655190 TI - [Results of the Ilizarov method in the treatment of pseudoarthrosis of the lower extremities]. AB - OBJECTIVES: We evaluated the results of the Ilizarov method in the treatment of pseudoarthrosis of the lower extremities. METHODS: Forty-six patients (34 men, 12 women; mean age 38.6 years; range 28 to 69 years) were treated by the Ilizarov method for femoral (n=8, 17%) and tibial (n=38, 83%) pseudoarthrosis. The mean duration of the disease was 1.6 years (range 6 months to 4.8 years). Pseudoarthrosis was hypertrophic in seven patients (16%) and atrophic in 39 patients (84%). The mean number of previous operations was 1.4 (range 0 to 4); the mean bone loss was 7.4 cm (range 3 to 12 cm); the mean shortening was 6.8 cm (range 0 to 12 cm); the mean size of the defect was 5.2 cm (range 3 to 12 cm). Applications were monofocal in 30 patients (66%) and bifocal in 16 patients (34%). The mean follow-up was 22.6 months (range 9 to 54 months). RESULTS: Union occurred in all patients (92%) but four (2 monofocal, 2 bifocal). The fixator was applied for a mean of 208 days (range 93 to 750 days), which was 162 days (range 98 to 296 days) for monofocal, and 286 days (range 140 to 496 days) for bifocal applications. According to the Paley's criteria, the results for bone healing and function were excellent in 26 and 25 patients, good in 12 and 14 patients, fair in four and three patients, and poor in four patients, respectively. Pin tract infections developed in 28 patients, and reflex sympathetic dystrophy in three patients. Refracture occurred after the removal of the frame in three patients who received bifocal treatment. One patient developed transient peroneal nerve palsy with drop foot. Equinus rigidity of the ankle was seen in four patients. Cancellous bone grafting was performed in four patients (25%) in whom delayed healing was observed at the target site following segmental bone transport. Three patients had union with a residual deformity of more than 7 degrees. In the monofocal group, none of the patients had a residual shortening of more than 1 cm. Following bifocal applications, no bone defects were observed; the mean residual length discrepancy was 1. 5 cm (range 0 to 4 cm), and the healing index was 52 days/cm. CONCLUSION: The Ilizarov technique may simultaneously be successful in the treatment of joint contractures, angular, rotational, and translational deformities, shortening, and bone defects. PMID- 12655191 TI - [Interlocking intramedullary nailing of humeral shaft fractures in adults]. AB - OBJECTIVES: We evaluated the effectiveness of interlocking intramedullary nailing in the treatment of humeral shaft fractures. METHODS: Forty-three patients (30 men, 13 women; mean age 42 years; range 20 to 83 years) were treated with interlocking intramedullary nailing for humeral shaft fractures. All fractures but one (type II open) were closed. Three patients had radial nerve, one patient had radial and ulnar nerve dysfunction. As all the nerve lesions were assessed as neuropraxia, no immediate surgical exploration was performed. Intramedullary nailing was performed by closed (18 fractures) and open technique (25 fractures), with (n=22) or without (n=21) reaming. Functional evaluation was made according to the Constant-Murley's shoulder scoring system. The mean follow-up was 16.5 months (range 5 to 40 months). RESULTS: Radiologically, union was achieved in all patients in a mean of 15.6 weeks. No surgery-associated nerve injuries or infections were encountered. Preoperative nerve dysfunction resolved in all patients in a mean of 12.5 weeks. No signs of rotator cuff impingement or restriction were detected in two patients in whom the nails were placed high in close proximity to the shoulder girdle. Of these, one patient underwent reoperation for the removal of the nail after the fracture union completed. Functional results were excellent in all patients at the end of postoperative three months. No significant correlation was found between the techniques employed and time to union and functional outcome. CONCLUSION: Interlocking intramedullary nailing provides adequate fixation and early mobilization, and results in satisfactory radiographic and functional results in the treatment of humeral shaft fractures. PMID- 12655192 TI - [The use of a modified Weber technique for high tibial osteotomy]. AB - OBJECTIVES: We evaluated fixation in high tibial osteotomy with the use of a modified Weber technique (MWT), together with its advantages. METHODS: Thirty nine patients (40 knees) underwent high tibial osteotomy for varus deformities of the knee joint and medial compartment degeneration. Fixation was performed by a modified Weber technique in which a semi-tubular plate and a long leg screw were used in place of the original Weber instrumentation. Thirty-five patients were women and four were men, with a mean age of 52.5 years (range 25 to 67 years). The patients were evaluated before surgery both clinically and by HSS scores and the advantages and disadvantages of the technique were assessed. The mean follow up was 22 months (range 8 to 45 months). RESULTS: All patients were allowed to give weight on the affected leg to the extent of utmost tolerability on the forth postoperative day. At the end of two weeks, 60% of the patients could bear half of the body weight. Screw breakage occurred in one patient due to faulty application of the technique. Pseudoarthrosis was seen in none of the patients, nor were there any early or late complications. Radiologic evidence of union was observed in all osteotomy sites. All patients achieved full extension except two who had a flexion deformity of 5 degrees. The mean knee flexion was 130.2 degrees. CONCLUSION: High tibial osteotomy may provide a rigid fixation, postoperative early motion and low complication rates in varus deformities and medial gonarthrosis of the knee. Our data suggest that MWT results in good stability both statically and dynamically in the fixation of high tibial osteotomy. PMID- 12655193 TI - [Surgical treatment of scapular fractures]. AB - OBJECTIVES: To evaluate the findings and results in patients who were treated surgically for scapular fractures. METHODS: Six patients (4 females, 2 males; mean age 40 years; range 25 to 59 years) with scapular fractures were treated by open reduction and internal fixation. All the fractures were caused by traffic accidents and all the patients had associated injuries. Four patients had scapular neck fractures, three of whom had accompanying ipsilateral clavicular fractures. In two patients, the fractures involved the glenoid fossa. The mean follow-up period was 27 months (range 18 to 38 months). Functional evaluations were made according to the scoring system described by Herscovici et al. RESULTS: The results were excellent in six patients, good in one, and fair in one patient. Four patients had no pain. Three patients returned to preinjury jobs. Two patients who were housewives were able do their housework. The range of movement was greater than 120 degrees in both abduction and flexion, and the muscle strength was 5 in four patients. No postoperative complications were encountered. CONCLUSION: Open reduction and stabilization followed by early rehabilitation may be successful in preventing stiffness, pain, and disability in scapular neck fractures accompanied by ipsilateral clavicle fractures, or fractures involving the glenoid fossa and leading to significant displacement. PMID- 12655195 TI - [An arthroscopic study on the anterior intermeniscal ligament]. AB - OBJECTIVES: An arthroscopic study was made on the presence and anatomic features of the anterior intermeniscal ligament with regard to its thickness and attachment patterns to the anterior horn of the menisci, together with its relationship with associated non-traumatic intraarticular disorders of the knee. METHODS: This multicenter study included 136 consecutive patients (80 males, 56 females; mean age 40 years; range 15 to 74 years) in whom the presence of anterior intermeniscal ligament was sought during arthroscopic examination of the knee. Arthroscopic features of the ligament was evaluated by probe. RESULTS: The anterior intermeniscal ligament was detected in 80 knees (58.8%). Its presence did not differ significantly with respect to sex (33 females, 58.9%; 47 males, 58.8%; p>0.05). A cord-like appearance was observed in 19 cases (14%), whereas a membranous-like structure was identified in 61 cases (44.9%), 34 (55.7%) of which could only be detected following synovial abrasion. Attachment patterns according to the Nelson and LaPrade's description showed type A in 47 cases (58.8%), type B in 23 cases (28.8%), and type C in 10 cases (12.5%). Type A was more common among cord-like intermeniscal ligaments, although this did not reach significance (p>0.05). No significant association was found between the presence of the ligament and intraarticular disorders. The detection of discoid lateral meniscus was significantly associated with the presence of a cord-like intermeniscal ligament (p<0.05). Of eight cases with discoid lateral meniscus (5 complete, 3 incomplete), five patients exhibited a cord-like anterior intermeniscal ligament. CONCLUSION: This is the first arthroscopic study to define the anatomy and the incidence of the anterior intermeniscal ligament. Functional features of the ligament should be more clearly demonstrated biomechanically. PMID- 12655194 TI - [The correlation between the accuracy of steroid injections and subsequent shoulder pain and function in subacromial impingement syndrome]. AB - OBJECTIVES: We assessed the relationship between proper placement of corticosteroid injections and subsequent shoulder function and pain in subacromial impingement syndrome. METHODS: The study included 48 patients (29 women, 19 men; mean age 46.5 years; range 23 to 58 years) with subacromial impingement syndrome, whose complaints of shoulder pain lasted more than two months. To monitor the site of injection, contrast material was added to a mixture of steroid and local anesthetic solution. Injections were delivered into the subacromial bursa by an anterolateral approach. Radiographs of the joint were taken immediately afterwards to ensure the accurate placement of the injection. Shoulder function and pain were evaluated by visual pain scale, range of movement of the joint, and Constant scores before treatment, and half an hour and two weeks after the injections. RESULTS: The injections were placed accurately in 42 patients (87%), while in six patients (12.5%), delivery to the target site failed. Statistically significant improvements were observed in both groups half an hour after the injections (p<0.05). However, two weeks after the treatment evaluations showed that failure to obtain an accurate placement was associated with return to pretreatment values, while significant improvement continued in the other group. CONCLUSION: Failure to deliver injections to the target site may be decreased by increased utilization of visualization and imaging methods. PMID- 12655196 TI - [Early results of complete subtalar release in congenital clubfoot deformity]. AB - OBJECTIVES: We evaluated the radiological and clinical results of complete subtalar release in patients with congenital clubfoot deformity. METHODS: We performed complete subtalar release in 30 feet of 20 patients (14 boys, 6 girls; mean age 8 months; range 3.5 to 18.5 months). We employed the Cincinnati incision in all cases. The results were evaluated according to the Simon's clinical and radiological criteria. Tarsal bone problems were analyzed separately. The mean follow-up period was 24.6 months (range 12 to 41 months). RESULTS: Clinical results were satisfactory in 24 feet (80%) and unsatisfactory in six feet (20%). Radiologically, satisfactory and unsatisfactory results were encountered in 19 feet (63%) and 11 feet (37%), respectively. Undercorrection occurred in 13 feet (43.3%) and overcorrection in one foot (1.4%). Revision surgery which was recommended for five undercorrected feet (16.6%) with unsatisfactory clinical and radiological results could be performed in four feet (13.3%). Deep skin necrosis that occurred in two feet (one with Achilles tendon necrosis) was treated surgically. Superficial skin necrosis developed in five feet and soft tissue infection in one feet, all of which responded well to conservative treatment. Other complications included talar head hypoplasia and navicular hypoplasia in two feet, respectively. CONCLUSION: Complete subtalar release allows to correct all components of clubfoot deformity in a single session. The results may appreciably be improved with enhanced surgical experience. PMID- 12655197 TI - [The injection of acrylic bone cement prevents bone collapse in the intercalar bones lacking bony support: an experimental sheep semilunar bone model]. AB - OBJECTIVES: In a sheep semilunar bone model, we investigated whether collapse in the intercalar bones lacking bony support could be prevented by the injection of acrylic bone cement. METHODS: The study included 16 limbs of eight sheep. Preoperatively, anteroposterior and lateral views of the carpal joints in the fore limbs were obtained. The animals were divided into four groups. In group 1 (n=3) no surgical procedure was performed in the right semilunar bones, whereas the periosteum on the contralateral side was elevated (group 2; n=3). The first two groups were left as controls. In Group 3 (n=5) the left semilunar bones were filled with acrylic bone cement following decancellation of the bone, while the right semilunar bones were left decancellated (group 4; n=5). The sheep were monitored for three months. Radiographs of the carpal joints were obtained to evaluate collapse occurrence in the semilunar bones. Thereafter, the animals were sacrificed and the semilunar bones were excised for biomechanical and histological examinations. Osteonecrosis and cartilage damage were sought and resistance to compressive forces was investigated. RESULTS: Radiologically, the extent of collapse was statistically significant in the semilunar bones in group 4 (p<0.05). The use of acrylic bone cement was found to prevent collapse in group 3, with no significant difference being noted between preoperative and postoperative semilunar bone heights (p>0.05). Biomechanically, the least resistance to compressive forces was measured in group 4 (p<0.05). Histologically, cartilage damage and osteonecrosis were only seen in group 4. CONCLUSION: Our data suggest that the use of acrylic bone cement prevents collapse in the semilunar bones, without inducing any cartilage damage or osteonecrosis. PMID- 12655198 TI - [One-bone forearm operation for defective ulna osteomyelitis: a case report]. AB - A four-year-old boy with forearm instability secondary to defective ulna osteomyelitis underwent one-bone forearm operation in neutral supination pronation position. A year after the operation, shortening (3 cm) of the forearm was detected. Radiographs showed delayed union; however, a stable and functional forearm was obtained. The patient was able to use his fingers. PMID- 12655199 TI - [Suprascapular nerve entrapment by a ganglion cyst: a case report]. AB - A twenty-two-year-old male patient presented with complaints of muscle atrophy and shoulder pain. Clinical and radiological studies showed a ganglion cyst near the spinoglenoid notch and a diagnosis of compression of the inferior branch of the suprascapular nerve was made. Following open excision of the cyst by a posterior approach, the patient's complaints subsided. A year after the operation recurrence of the lesion was detected. Arthroscopic control showed no intrarticular lesion. The patient still experiences minimal pain from time to time. PMID- 12655200 TI - [Total calcanectomy in a patient with a giant cell tumor]. AB - Total en-bloc calcanectomy was performed in a 36-year-old male patient with a giant cell tumor in his right calcaneus. A year after surgery, the patient had no complaints, and a good functional result was obtained. He was able to return to his work. No local or metastatic recurrences were encountered. PMID- 12655201 TI - [Tuberculous osteitis of the tarsal bones: a case report]. AB - A thirty-year old woman presented with complaints of pain, swelling in her right ankle, and difficulty in walking that had a history of six months. The range of motion of the joint was limited. She did not get any benefit from various non steroidal anti-inflammatory drugs. She had undergone surgery three years ago for pain in her right hip, difficulty in walking, and a swelling in the right inguinal area. She had an elevated erythrocyte sedimentation rate. Foot radiographs showed degeneration of the navicular and cuneiform bones. Hyperintensity in the same area on magnetic resonance scans was interpreted in favor of a chronic infection. Arthrotomy of the calcaneocuboid joint and the 3rd, 4th, and 5th tarsometatarsal joints was performed, with curettage of the cuboid and lateral cuneiform bones. Tissue samples were obtained from the synovium, joint capsule, and bone, among which Mycobacterium tuberculosis was recovered only in bone cultures. The patient manifested rapid improvement with antituberculous therapy. This case demonstrates the need to obtain cultures from the bone as well as those from the synovial fluid or tissues to identify tuberculosis of a very rare site of involvement. PMID- 12655202 TI - Maxillary volume growth in childhood. AB - Nasomaxillary abnormalities in form, position, and development in children are often prominent features of craniosynostosis, and in particular, craniofacial dysostosis. While attempting to quantitatively assess the volumetric maxillary deficiency in these patients, it became apparent that there was no "normal" reference range for maxillary volumes throughout childhood that could be used for comparison. The aim of this study was to generate a model for measuring maxillary volume and subsequent changes throughout childhood. The technique of segmentation was applied to magnetic resonance images obtained in 55 healthy children (30 boys, 25 girls), aged 1 month to 184 months (15.33 years). Maxillary volumes were plotted against age for boys and girls to create a model for normal maxillary growth during the first 15 years of life. Maxillary volumes were larger in boys at all ages. However, the pattern of maxillary growth in boys and girls was similar and could be divided into three periods, each lasting approximately 5 years. During the first 5 years of life, there is a steady increase in maxillary volume, at the end of which the maxilla has reached 53 percent of the volume recorded at 15 years. There is an accelerated rate of growth between 5 and 11 years, which corresponds to the development and eruption of the permanent dentition. Thereafter, until the age of 15 years, the rate of growth of the maxilla plateaus. Maxillary volume in the first 12 months of life is, on average, 29 cm3 in boys and 25 cm3 in girls. By 15 years of age, it has increased to an average of 73.0 cm3 in boys and 59.4 cm3 in girls (an increase by a factor of 2.5 in boys and 2.4 in girls). The difference between the two sexes is statistically significant for the entire series (boys: mean maxillary volume = 56.55 cm3, SD = 24.61; girls: mean maxillary volume = 40.68, SD = 17.69, p = 0.009, one-way analysis of variance). PMID- 12655203 TI - Maxillary volume growth in craniosynostosis. AB - Craniosynostosis, and in particular, craniofacial dysostosis, exhibits abnormalities of the nasomaxillary complex in form, position, and development. The aim of this study was to quantitatively assess the volumetric maxillary abnormality in patients at the time of initial diagnosis of craniosynostosis and to make comparisons with a "normal" reference range for maxillary volumes throughout childhood. The technique of segmentation was applied to preoperative computed tomographic head scans obtained in 31 children (14 boys, 17 girls), between 1 and 34 months of age (mean, 11.06 months), who underwent cranial expansion surgery for craniosynostosis affecting the coronal suture complex. Maxillary volumes were plotted against age for the first 3 years of life and were compared with a healthy population. There was no statistical difference between the two sexes for mean maxillary volume. The mean maxillary volumes for the entire group were statistically smaller than the norm (p = 0.046, linear regression with age as a covariable), but there was no statistical difference among the four different groups of coronal synostosis (unilateral coronal, nonsyndromic bilateral coronal, nonsyndromic complex pansynostosis, syndromic bilateral coronal synostosis) (p = 0.407, one-way analysis of variance). On graphic data analysis, the maxillary volume was smaller than the norm in craniosynostotic children who presented in the first few months of life. However, by 7 months of age in nonsyndromic bilateral coronal synostosis and by 17 months of age in syndromic bilateral coronal synostosis, the maxillary volumes had increased toward the norm. This implies that the effect of the craniosynostotic process on the midface structures is present from birth and parallels the effect on the cranial vault sutures. PMID- 12655204 TI - Detection of subclinical infection in significant breast implant capsules. AB - The pathogenesis of fibrous capsular contracture after augmentation mammaplasty is still debated. One hypothesis implicates low-grade bacterial infections as a cause. The presence of a staphylococcal biofilm in a patient with recurrent capsular contracture was previously reported. A comparative, prospective, blinded, clinical study of implants and capsules removed from patients with or without significant capsular contracture was conducted to investigate the association of biofilm contamination, breast implants, and capsular contracture. Capsule and implant samples obtained during explantation were tested by routine microbiological culture, sensitive broth culture (after maceration and sonication), and scanning electron microscopy. Clinical parameters were correlated with microbiological findings. A total of 48 implant and/or capsule samples were obtained from 27 breasts during a 22-month period. Of the 27 breasts, 19 exhibited significant contracture (Baker grade III/IV). The mean duration of implantation was 9.2 years (range, 0.4 to 26.0 years). Routine swab cultures obtained at the time of explantation were negative for bacterial growth for all samples. The sensitive broth culture technique yielded 24 positive samples (50 percent, n = 48). An analysis of capsules demonstrated that 17 of 19 samples obtained from patients with significant contracture were positive, compared with only one of eight samples obtained from patients with minimal or no contracture (p = 0.0006). Fourteen of the 17 positive cultures from significantly contracted breasts yielded coagulase-negative staphylococci, mainly, species of the Staphylococcus epidermidis group. The presence of coagulase-negative staphylococci was also significantly associated with capsular contracture (p = 0.01). There was no significant difference in the frequency of culture positivity for saline versus silicone implants (p = 0.885). Scanning electron microscopy confirmed the presence of extensive biofilm on implants and within capsules.Biofilm, in particular, S. epidermidis biofilm, was detected for a significant proportion of patients with capsular contracture. This implicates biofilm disease in the pathogenesis of contracture, and strategies for its prevention should be explored. PMID- 12655205 TI - Surface landmarks to locate the thenar branch of the median nerve: an anatomical study. AB - The thenar branch of the median nerve can be injured during carpal tunnel release. The purpose of this study was to identify surface landmarks to consistently predict the location of the thenar branch of the median nerve. Surface landmarks were marked and incised in 28 cadaveric hands. The incisions were made along the longitudinal line of the third web space and the horizontal cardinal line from the hamate hook to the ulnar border of the thumb. The origin of the thenar branch was determined in relation to these longitudinal and horizontal vectors. The origin of the thenar nerve branch was consistently observed in the radial proximal quadrant formed by the aforementioned longitudinal and horizontal vectors. The thenar branch origin was observed to be an average of 8.6 +/- 1.9 mm radial to the longitudinal axis along the third web space. The origin of the thenar branch was observed to be an average of 6.3 +/- 2.0 mm proximal to the horizontal axis between the hamate hook and the ulnar border of the thumb. The thenar branch was observed precisely at the intersection of the longitudinal vector from the second web space to the scaphoid tubercle and the horizontal vector from the hamate hook to the radial edge of the proximal metacarpophalangeal crease in all 28 cadaveric hands. On the basis of these 28 cadaveric dissections, the location of the thenar branch of the median nerve can be predicted by the intersection of the longitudinal vector from the second web space to the scaphoid tubercle and the horizontal vector from the hamate hook to the radial aspect of the metacarpophalangeal crease. PMID- 12655206 TI - Open carpal tunnel release using a 1-centimeter incision: technique and outcomes for 104 patients. AB - The advantages of endoscopic carpal tunnel release, compared with traditional open techniques, include smaller incisions, less scar tenderness, and faster recoveries. However, endoscopic carpal tunnel release has also been associated with higher complication rates. The goal of this study was to evaluate the safety and functional outcomes of minimal-incision open carpal tunnel release. In this prospective study involving a 2-year period, 104 patients (149 hands) underwent open carpal tunnel release with a 1-cm incision. Prospective data on complications among 104 patients were recorded, and functional outcomes among 20 patients were assessed by using the Michigan Hand Outcomes Questionnaire, the Jebsen-Taylor Hand Function Test, and pinch/grip strength testing. Data were collected before the operation and 3 weeks and 6 months after the operation. Complications included three wound infections and one carpal tunnel syndrome recurrence, 18 months after the initial release procedure. Michigan Hand Outcomes Questionnaire scores improved significantly between the preoperative and postoperative periods. There were no significant changes in Jebsen-Taylor Hand Function Test results or pinch/grip strength. Minimal-incision open carpal tunnel release can be performed safely and is associated with good functional outcomes. PMID- 12655207 TI - Treatment of complex interphalangeal joint fractures with dynamic external traction: a series of 20 cases. AB - Data are reported for a series of 20 patients who were treated with the pins and rubbers traction system for fractures of the proximal interphalangeal joints of the long fingers. This technique allows fracture reduction with external dynamic traction and immediate active mobilization. Two patients in the series were lost to follow-up monitoring. For two others, the pins and rubbers traction system needed to be removed early (during the first week) because of intolerance or infection. Sixteen patients who were reexamined after minimal follow-up periods of 1 year demonstrated a mean active range of motion of 85.9 degrees for the injured joint; only one patient experienced intermittent pain. PMID- 12655208 TI - Anterolateral thigh flap in the treatment of postburn flexion contractures of the knee. AB - The use of the anterolateral thigh fasciocutaneous flap in the reconstruction of soft-tissue defects around the knee among burn patients is described. The anterolateral thigh fasciocutaneous flap was used for eight patients (all male; mean age, 45 years; age range, 32 to 60 years). Flexion contracture was observed for seven patients with unhealed wounds and one patient with a healed burn wound. The anterolateral thigh flap was used as a free flap for six patients and as a distally based island flap for two patients. The flaps ranged from 8 to 17 cm in width and from 12 to 30 cm in length. Seven flaps were based on a musculocutaneous perforator, and two of them were thinned before transfer to the defect. A true septocutaneous perforator was observed in only one case. The mean follow-up period was 12.5 months (range, 3 to 23 months). Only one flap exhibited distal superficial necrosis, which did not compromise the final result. All patients returned to ambulatory status in 15 to 22 days. Extensor splints were applied to prevent mobilization of the skin graft at the flap donor site for only 7 days. The anterolateral thigh flap has many advantages for the reconstruction of postburn flexion contracture of the knee, as follows: (1) very large thin flaps can be elevated, (2) the two-team approach is possible, (3) color and texture matches are good, (4) the donor-site scar can be easily hidden, and (5) the technique allows early mobilization and patients can return to normal daily activity in a short time. Free or distally based anterolateral thigh flaps are a good choice, both aesthetically and functionally, for the reconstruction of soft tissue defects of the knee region. PMID- 12655209 TI - Induction of vascular endothelial growth factor by fibrin as a dermal substrate for cultured skin substitute. AB - In the initial phase of wound healing, endogenous fibrin clots are known to form a provisional matrix and to promote angiogenesis. Growth factors such as vascular endothelial growth factor (VEGF) increase in wounds to stimulate angiogenesis. However, it remains unknown whether VEGF is induced when fibrin is used as a dermal substrate for cultured skin substitutes. The authors investigated the effect of fibrin gel as a dermal substrate for a cultured skin substitute, using human keratinocytes and dermal fibroblasts. A collagen-cultured skin substitute was also examined for comparison. VEGF in the culture supernatant in both types was measured by enzyme-linked immunosorbent assay, and VEGF mRNA was determined semiquantitatively by reverse-transcriptase polymerase chain reaction after 2 days of incubation. Experiments were performed using 12 cultured skin substitutes: four for histologic examination before transplantation, four for VEGF assay in vitro, and four for the transplantation to athymic mice. Three independent experiments were performed for each step. VEGF concentration in the fibrin-cultured supernatant was 84.3 +/- 11.8 pg/ml, whereas it was 27.8 +/- 4.68 pg/ml in the case of the collagen substrate. The relative levels of VEGF mRNA were 1.088 +/- 0.100 and 0.698 +/- 0.226, respectively. In in vivo transplantation, the fibrin-type cultured skin substitute showed an excellent take on the wound bed, and a normally proliferating keratinocyte layer with emergence of vascular endothelial cells in the transplanted floor was seen 3 days after transplantation. Vascular endothelial cells, which were identified using alkaline phosphatase stain, were significantly increased in the fibrin-type cultured skin substitute. The use of fibrin as a dermal substrate for cultured skin substitute increases the secretion of VEGF, improves regeneration of mature epidermal structure after in vivo transplantation, and promotes the migration of vascular endothelial cells. PMID- 12655210 TI - Evaluation of the hormonal function and histological features of heterotopic isogenic ovarian transplantation in rats. AB - The purpose of the study was to determine the feasibility of preserving ovarian function after heterotopic transplantation by means of microvascular anastomosis of the transplanted vascular pedicles to a set of preselected vessels. Six groups of 10 Sprague-Dawley inbred rats were used in this study. Group I underwent bilateral ovariectomy operation and served as the ovariectomy control. Group II underwent bilateral ovariectomy followed by heterotopic isogenic ovarian implantation. Group III underwent bilateral ovariectomy and isogenic heterotopic ovarian transplantation by means of microvascular anastomosis. Group IV served as the laparotomy sham-operated control. Group V served as the ovarian donor for group II. Group VI served as the donor of the ovarian-kidney vascular pedicle complex for group III. Postoperative ovarian estradiol levels were measured, and histological characteristics were elucidated in groups I, II, III, and IV. The results demonstrated that the estradiol level of the transplantation group was comparable to that of the sham operation group and was significantly higher than that of the implantation group. Histologically normal ovarian architecture was observed in the sham group (IV) and also in the transplantation group (III). Altered architecture was observed in the implantation group (II). These findings indicate that extraabdominal heterotopic ovarian transplantation with microvascular anastomosis led to normal ovarian hormonal function and was effective in preserving oocyte production capacity. PMID- 12655211 TI - Effect of single-dose administration of FK-506 on allogeneic limb transplantation in rats: is there an optimal timing of its administration? AB - Although a number of experimental studies have demonstrated that high-dose administration of FK-506 induces the success of limb allogeneic transplantation in rats, some of them have reported occurrences of lethal side effects. Therefore, a more effective regimen with lower-dose administration of this agent must be developed. The objective of this study was to determine an optimal timing of a single-dose administration of FK-506 in rat limb allografts. In the current study, orthotopic hindlimb transplantations were performed using major histocompatibility mismatched pairs of inbred rats. The rats were classified into five groups on the basis of the different time schedules of FK-506 administration as follows: syngeneic group, Lewis-to-Lewis transplantation; control group, ACI to-Lewis without any immunotherapy; day 0 group, recipients treated with a single dose administration of FK-506 (5 mg/kg of body weight, intramuscular injection) at day 0 postoperatively; day 1 group, with a single-dose administration of FK 506 at day 1 postoperatively; and day 2 group, with a single-dose administration of FK-506 at day 2 postoperatively. The median graft survival time in each group (n = 5) was 9 days in the control group, 19 days in the day 0 group, 49 days in the day 1 group, and 42 days in the day 2 group. The values in the day 1 and the day 2 groups significantly increased compared with those in other groups. For prolonged survival of a grafted limb, a single-dose administration of FK-506 is more effective at 24 to 48 hours after transplantation. PMID- 12655212 TI - Low-molecular-weight heparin (dalteparin) effectively prevents thrombosis in a rat model of deep arterial injury. AB - Unfractionated heparin is often used to prevent thrombosis in microvascular surgery, but a major drawback of heparin therapy is increased bleeding. Low molecular-weight heparins prevent venous thrombosis as effectively as heparin and have better bioavailability and a longer plasma half-life, which explains the increased use of low-molecular-weight heparins as substitutes for heparin in clinical practice. However, the ability of low-molecular-weight heparins to prevent arterial thrombosis has been debated. In this study, the authors compared the antithrombotic and antihemostatic effects of heparin and the low-molecular weight heparin dalteparin in a rat model of arterial thrombosis. A segment of the left common carotid artery was isolated between vascular clamps and opened longitudinally. An endarterectomy was performed and the arteriotomy was closed with a running suture. The antithrombotic effect (vascular patency 31 minutes after reperfusion) and the surgical bleeding were measured. Groups of 10 rats were treated in a blind, random fashion with intravenous injection of one of the following substances 1 minute before clamp release. Three groups received a bolus of heparin (20, 60, or 180 IU anti-factor Xa/kg), three groups received dalteparin (60, 180, or 540 IU anti-factor Xa/kg), and one group was treated with vehicle (saline). Heparin 180 IU/kg produced a distinct antithrombotic effect compared with the control group (p = 0.03), but it also significantly increased the surgical bleeding to 2.0 g compared with 1.5 g in the control group (medians, p = 0.01). Dalteparin 180 and 540 IU/kg also produced a powerful antithrombotic effect (p = 0.01 and p = 0.03, respectively). In contrast to heparin, 180 IU/kg dalteparin did not increase the surgical bleeding (median, 1.5 g; p = 0.37 versus controls). Dalteparin 540 IU/kg increased the median surgical bleeding to 2.6 g (p = 0.06 versus controls). The nonsignificant difference may be explained by the great interindividual variation of surgical bleeding in the high-dose dalteparin group. Dalteparin prevented arterial thrombosis as effectively as unfractionated heparin. In contrast to heparin, dalteparin did not increase the surgical bleeding, which indicates that dalteparin instead of heparin can be used to prevent thrombosis in microvascular surgery. PMID- 12655213 TI - Assessment of the effects on growth of porous hydroxyapatite granule cranioplasty in the immature guinea pig craniofacial skeleton. AB - The immature guinea pig was used to study the effects on growth of porous granular hydroxyapatite used as an onlay cranioplasty and inlay cranioplasty to reconstruct full-thickness cranial defects in a growing craniofacial skeleton. Forty Hartley guinea pigs, 20 immature animals and 20 mature animals, were divided into four groups each containing five mature and five immature animals. The mature animals served as controls. Group I underwent elevation and replacement of the parietal periosteum. Group II underwent placement of hydroxyapatite between periosteum and parietal bone. Group III underwent elevation and replacement of autogenous bone flap after the formation of a 1 x 1 cm craniectomy defect in the parietal skull. Group IV underwent elevation of a 1 x 1-cm parietal craniectomy and reconstruction of the defect with hydroxyapatite granules placed between the dura and periosteum. Immature animals were killed at maturity at 3.5 months and mature animals were killed 2.5 months postoperatively. Macroscopic examination of the operative field, transverse and longitudinal cephalometric measurements, and histological sections encompassing the operative sites were compared. Macroscopically, all reconstructed operative sites were fully incorporated into the cranium. Histological staining of the sectioned operative site revealed no hydroxyapatite migration through the cranial bone or dura. No inflammatory or foreign body reaction was evident in the subcutaneous tissue, periosteum, or dura. No statistically significant cephalometric intergroup or intragroup differences were found at the conclusion of the study. The results of this study indicate that a granular porous form of hydroxyapatite may be used as an onlay or inlay cranioplasty in the immature guinea pig craniofacial skeleton without evidence of dural inflammation, granule migration, or growth restriction or retardation. PMID- 12655214 TI - Correction of warfarin-induced nasal hypoplasia. PMID- 12655215 TI - A prefabricated osteocutaneous flap for tracheal reconstruction. PMID- 12655216 TI - Use of waxing screws for accurate primary placement of endosteal implants in the vascularized fibular bone-reconstructed mandible. PMID- 12655217 TI - The extended latissimus dorsi flap revisited. PMID- 12655218 TI - A history of noma, the "Face of Poverty". AB - Noma (necrotizing ulcerative stomatitis, stomatitis gangrenosa, or cancrum oris) is a devastating orofacial gangrene that occurs mainly among children. The disease has a global yearly incidence of 140,000 cases and a mortality rate of approximately 90 percent. Patients who survive noma generally suffer from its sequelae, including serious facial disfigurement, trismus, oral incontinence, and speech problems. The medical history of noma indicates that the disease was already known in classical and medieval civilizations in Europe. In the sixteenth and seventeenth centuries, Dutch chirurgeons clearly described noma as a clinical entity and realized that the popular name "water canker" was not sufficient, because this quickly spreading ulceration in the faces of children was different from "cancer." In the eighteenth century, awareness that noma is related to poverty, malnutrition, and preceding diseases such as measles increased in northwestern Europe. In the first half of the nineteenth century, extensive surgical procedures were described for the treatment of the sequelae of noma. At the end of that century, noma gradually disappeared in the Western world because of economic progress, which gave the poorest in society the opportunity to feed their children sufficiently. Only in the twentieth century were effective drugs (sulfonamides and penicillin) against noma developed, as well as adequate surgical treatment for the sequelae of noma. These modes of treatment remain inaccessible for the many present-day victims of noma because of their extreme poverty. The only truly effective approach to the problem of noma throughout the world is prevention, namely, combating the extreme poverty with measures that lead to economic progress. In the meantime, medical doctors in the Western world should not forget their own history and ignore this global health problem; rather, they should face "the face of poverty" with the eyes of mercy and concern suited to their profession. PMID- 12655219 TI - Subciliary versus subtarsal approaches to orbitozygomatic fractures. AB - LEARNING OBJECTIVES: After studying this article, the participant should be able to: 1. Describe the anatomic differences in the subciliary versus the subtarsal approach. 2. Discuss the difference between the "skin-only" and the "skin-muscle flap" variations of the subciliary approach. 3. Discuss the potential complications of both approaches. 4. Discuss the advantages of the subtarsal approach versus the subciliary approach. Many incisions have been described for approaches to orbitozygomatic fractures, the most frequently used being the subciliary incision with its modifications, the subtarsal incision, and the transconjunctival incision with or without lateral canthotomy. Each of these approaches has its advantages and disadvantages that may make it more or less appealing to use depending on the patient's age and severity of fracture. A balance must be struck between adequate exposure and acceptable cosmetic result. This article reviews the literature with particular respect to the transcutaneous approaches of subciliary versus subtarsal techniques in the treatment of orbitozygomatic fractures. PMID- 12655220 TI - Plastic surgery knowledge: state of the art. PMID- 12655221 TI - The bare area of the upper conjunctiva: a closer look at the anatomy of transconjunctival upper blepharoplasty. AB - Transconjunctival upper blepharoplasty is a novel technique for which the overall experience has been limited. The authors have used this technique in 42 patients for eyelid rejuvenation, with predictable results and low morbidity. The authors previously described the "bare" area in the medial upper conjunctiva, which was initially noted during their clinical series. This area serves as an anatomic window to access the medial upper eyelid fat pad with a high degree of safety. To clarify the anatomy of this approach, an anatomic study of the upper medial eyelid was performed on the orbits of 10 cadavers (20 orbits). The bare area was further elucidated during these dissections and its microscopic components described in detail. Its spatial relationship to the medial orbital wall and trochlea was also examined during this study. The authors hope that these findings will aid surgeons planning to use this technique. PMID- 12655223 TI - Aesthetic and restorative midface lifting with hand-carved, expanded polytetrafluoroethylene orbital rim implants. AB - The midface lift represents an important advance in aesthetic and reconstructive surgery. However, the need for reliable fixation along the orbital rim has been a significant challenge. Furthermore, volume is needed at the orbital rim, to compensate for long-term remodeling of the bone of the orbital rim and malar face. A technique using a hand-carved, expanded polytetrafluoroethylene implant that is permanently anchored to the orbital rim with titanium microscrews, creating a site for fixation of the advanced midface soft tissues, was developed. This report presents a retrospective, uncontrolled, case series of 41 consecutive patients who underwent transconjunctival midface operations with these implants, and it addresses a variety of midface aesthetic and reconstructive deficits. Only patients with at least 6 months of follow-up data were included in the study. To date, significant complications have been limited. The complications included two cases of implant palpability, with only one requiring surgical modification. One patient underwent implant removal because of skin breakdown and infection related to recurrent squamous cell carcinoma. One patient required revisional lateral canthoplasty for reasons of symmetry. On the basis of this series, hand-carved, expanded polytetrafluoroethylene implants seem to have significant advantages, compared with previously available orbital rim implants. These advantages include the ability to easily modify the implant for the individual anatomical needs, the creation of a secure anchor for fixation of advanced midface soft tissues, excellent tolerance of the implant material, and the ability to place the implant with limited exposure. The greatest disadvantage is the need for the surgeon to carve the implant, which requires time and carving skill. Despite this limitation, the technique is promising. PMID- 12655222 TI - The safe face lift with bony anatomic landmarks to elevate the SMAS. AB - The risk for facial nerve injury has been reported to be increased with the inclusion of superficial musculoaponeurotic system (SMAS) elevation as compared with a skin-only face lift. The facial nerve courses through the parotid gland. The SMAS is elevated superficial to the parotid gland. However, in elevating the SMAS anterior to the parotid gland, the facial nerve is at risk of injury where its branches emerge from the anterior edge of the parotid gland. The purpose of this study was to identify bony anatomic landmarks to predict the location of the anterior edge of the parotid gland to avoid injury to the facial nerve branches as they exit the parotid gland. The authors dissected 20 cadaver face halves to determine bony landmarks-the masseteric tuberosity and the inferior lateral orbital rim-to predict the location of the anterior parotid edge. Then they measured the anterior edge of the parotid gland in relation to the vector formed between these two bony landmarks. They identified and measured the most anterior portion of the parotid gland in relation to this vector. Then the most posterior aspect of the parotid gland in relation to this vector was measured. In the 20 dissections, the authors found the most anterior portion of the parotid gland to be 2.7 +/- 1.0 mm anterior to the vector from the inferior lateral orbital rim to the masseteric tuberosity. The most posterior part of the anterior edge of the parotid gland in relation to this vector was found to be 1.0 +/- 1.5 mm posterior to this vector. The parotid gland measured an average of 38.8 +/- 3.5 mm in width from the tragus to the anterior parotid edge. In elevating the SMAS with a face lift, the facial nerve branches can be predicted to exit the anterior edge of the parotid gland, which can be located 38.8 mm anterior to the tragus and near the vector from the inferior lateral orbital wall to the masseteric tuberosity. PMID- 12655225 TI - The HIPAA Privacy Rule: what it means for submissions to the Journal. PMID- 12655224 TI - Nondebridement of laser char after two carbon dioxide laser passes results in faster reepithelialization. AB - Skin repair following laser injury can be accelerated by using techniques that promote rapid reepithelialization. In this article, the benefit of intraoperative nondebridement of laser debris after two laser passes is discussed. After carbon dioxide laser resurfacing of the face, skin specimens were examined using indirect immunofluorescence with antibodies to specific epidermal and basement membrane proteins. Biopsy specimens obtained immediately after resurfacing showed a greater injury to epidermal and basement membrane proteins when skin was wiped with saline-soaked gauze after laser passes than when there was no debridement after two passes. Later examination of skin specimens obtained from nine patients 2 days after carbon dioxide resurfacing showed that nondebrided, occluded skin had faster reepithelialization than the other treatments. Nondebridement of the skin at the time of resurfacing along with the use of postoperative occlusive dressings led to the rapid reestablishment of a multilayered epidermis only 2 days after resurfacing. Nondebridement along with occlusive dressings results in rapid reepithelialization of the skin after two carbon dioxide laser passes for skin rejuvenation. PMID- 12655226 TI - What you can do is not what they want. PMID- 12655227 TI - Profiling. PMID- 12655228 TI - Training of plastic surgery on the white board. PMID- 12655229 TI - Simple and effective device for finger tourniquet: a rolled penrose drain. PMID- 12655230 TI - Why have paper when you can have silk? PMID- 12655231 TI - Atypical fibroxanthoma and metastasis to the lung. PMID- 12655232 TI - Do we sacrifice ourselves? PMID- 12655233 TI - An interesting mammary implant rupture: seat belt injury. PMID- 12655234 TI - Utilizing a surgical glove to trace skin defects on fingers. PMID- 12655235 TI - An easy way of finding a vein in microsurgery. PMID- 12655236 TI - Fate of diced cartilage grafts wrapped with oxidized regenerated cellulose. PMID- 12655237 TI - A novel use of the surgical cap. PMID- 12655238 TI - Digital blocks with or without epinephrine. PMID- 12655239 TI - Can sensorial nerve transection be distinguished with the "naked eye"? Clinical sign of the ulnar nerve transection. PMID- 12655240 TI - Clipped microvascular anastomoses in lower-leg free flaps. PMID- 12655241 TI - The levator septi nasi muscle and its clinical significance. PMID- 12655242 TI - Pedicled or free flap reconstruction of the oral cavity and tracheostomy tube management: the ring stitches. PMID- 12655243 TI - Minimal access cranial suspension lift. PMID- 12655244 TI - Gigantic metameric seborrheic keratosis. PMID- 12655245 TI - Faulty eudermic glove. PMID- 12655246 TI - The ex-suture: a nonbleeding excision for hair transplantation. PMID- 12655247 TI - An areola marker. PMID- 12655248 TI - Forehead lifting for cutis verticis gyrata. PMID- 12655249 TI - Two practical devices for monitoring temperature. PMID- 12655250 TI - A firm donor-site dressing. PMID- 12655251 TI - Cholangiopathy and the biliary cast syndrome. AB - Biliary casts are uncommon but are more frequently described in liver transplant patients. To our knowledge there have been only two published cases describing biliary casts in non-liver transplant patients. The aetiology of cast development is not fully known but is likely to be multifactorial with the presence of biliary sludge being a prerequisite for cast formation. Bile duct damage and ischaemia, biliary infection, fasting, parenteral nutrition, abdominal surgery and possibly other factors, are all thought to be implicated in cast pathogenesis via sludge development. Endoscopic management has been shown to be effective in a minority of cases and may act as a temporary measure in others but surgical removal of casts is usually necessary. With a greater understanding and improvement in liver transplant surgical techniques and the management of post operative complications, the development and severity of biliary sludge and casts have decreased. PMID- 12655252 TI - Intrafamilial transmission of hepatitis B virus in the eastern Anatolian region of Turkey. AB - OBJECTIVE: To determine the possible route of hepatitis B virus (HBV) transmission throughout a family setting. METHOD: Two hundred and forty family members (42 spouses, 32 offspring, 34 mothers, 35 fathers and 97 siblings) of 84 HBsAg carriers attending the Ataturk University Medical Faculty Hospital between July 2000 and May 2001, and 384 first time blood donors (control group) were screened prospectively for markers of HBV by using an ELISA. The chi-squared and Fisher's exact tests were used for statistical analysis. RESULTS: The prevalence of HBsAg and HBV infection among the family members (29.6% and 43.8%, respectively) was significantly higher than in the control group (9.6%, and 29.7%), P < 0.001. Among family members, the highest prevalences of HBsAg and HBV infection were in the husbands (70.0% and 90.0%, respectively), offspring (66.7% and 100.0%) of female index cases, and siblings (40.2% and 49.5%, respectively) of all index cases. Transmission of HBV to offspring was high in cases where both the parents were positive, but there was no difference in cases where only the mother or father was positive. Despite a high prevalence of HBsAg in the offspring of female index cases, HBsAg and anti-HBs seroprevalences in the mothers of index cases were not higher than the control group (P < 0.05). Thus, it appears that mothers do not have an important role in the acquisition of HBV infection. CONCLUSION: In the area studied, in addition to the sexual route, both parents-to-child and sibling-to-sibling horizontal transmission might be the main route of intrafamilial spread of HBV infection. PMID- 12655253 TI - Development of antinuclear antibodies and its clinical impact in patients with Crohn's disease treated with chimeric monoclonal anti-TNFalpha antibodies (infliximab). AB - BACKGROUND: Although the efficacy of infliximab in Crohn's disease (CD) has been demonstrated, its safety profile has yet to be established. Autoimmune adverse events such as human anti-chimeric antibodies and the development of antinuclear antibodies (ANAs) have been notified, but the true incidence and clinical relevance of the latter is still unknown. OBJECTIVE: To evaluate the changes in ANA status in CD patients treated with infliximab and the clinical evolution of those who are ANA positive. METHODS: The ANA status of 36 CD patients treated with infliximab was determined at baseline and 6 weeks after the initial infliximab infusion. Patients were followed up monthly. In the case of infliximab re-treatment, ANA status was again evaluated. Twenty-eight patients (78%) were treated concomitantly with immunosuppressants. RESULTS: Eight patients (22%) were ANA positive at baseline; none developed anti-double-stranded DNA antibodies (aDNAds) at week 6. Three of them were re-treated: there were increasing ANA titres in all cases and developing aDNAds in two. Only six of 28 patients who were ANA negative at baseline changed their ANA status at week 6, but none developed aDNAds. One of them was retreated showing a further increase in ANA titre and developing aDNAds at high titre. No patient presented lupus-like syndrome. CONCLUSIONS: Only a few CD patients treated with infliximab and immunosuppressants develop ANAs. This condition is not associated with aDNAds and/or lupus-like syndrome in the majority of cases. PMID- 12655254 TI - Drug utilization study in patients with Crohn's disease in Spain. AB - OBJECTIVE: To discover the therapeutic strategies for Crohn's disease used in Spain and to analyse the factors associated with the use of each treatment. DESIGN: Observational, cross-sectional study of patients with Crohn's disease who attended hospitals in Spain. METHODS: Two structured questionnaires were used, one completed by gastroenterologists about demographic clinical data and disease activity, and the other a telephone interview with the patients to obtain epidemiological and drug utilization data. RESULTS: A total of 635 patients from 39 hospitals were included. Aminosalicylates (77%) and corticosteroids (41%) were the most commonly used drugs. Monotherapy was used in 52% of the patients and was associated with less severe disease and relapse. Aminosalicylate use was associated with relapse and associated inversely with use of immunosuppressors. Corticosteroid use was associated with relapse, disease activity, and emergency visit frequency, and associated inversely with perianal disease. Use of immunosuppressors was associated with perianal disease, chronic active disease, and routine visit frequency, and associated inversely with aminosalicylate use. Antibiotic use was associated with relapse, perianal disease, chronic active disease and hospitalization. CONCLUSION: Aminosalicylates, corticosteroids and a combination of both drugs are the drugs used most frequently against Crohn's disease in Spain. Gender, age, clinical pattern and location were not associated with the type of drug used either in monotherapy or in combined therapy. PMID- 12655255 TI - Wireless capsule video endoscopy is a superior diagnostic tool in comparison to barium follow-through and computerized tomography in patients with suspected Crohn's disease. AB - BACKGROUND: The recently introduced wireless M2A capsule video endoscopy (CVE) has been demonstrated to be superior to barium follow-through and enteroscopy in diagnosing patients with occult blood loss and iron-deficiency anaemia. OBJECTIVE: To further investigate CVE in comparison to barium follow-through and entero-computerized tomography (CT) in establishing the diagnosis of patients with suspected Crohn's disease. DESIGN AND SETTING: The study was conducted in one academic hospital. Twenty patients with recurrent abdominal pain and/or weight loss or chronic diarrhoea underwent barium follow-through as their initial examination, followed by CVE (if there was no stricture) and entero-CT. The radiologist that performed the barium follow-through and entero-CT was blinded to the results of the CVE. A blinded reader who was unaware of the study objective diagnosed the results of the CVE. In most cases in which there was a discrepancy between examinations, colonoscopy and ileoscopy were performed. For each patient, the diagnosis and disease extent were recorded. RESULTS: Twenty patients (13 males, 7 females; mean age 31 years, range 20-57) were included in the study. Ninety-five per cent of the patients had abdominal pain, 75% had diarrhoea, and 65% had weight loss. The mean haemoglobin level of the group was 13.1 g% (range 10-15.5). Only 13 patients underwent entero-CT.CVE confirmed the diagnosis of Crohn's disease that was suspected by alternative methods in six of the 20 patients. CVE made the diagnosis of Crohn's disease in six patients that had normal barium follow-through or entero-CT. CVE ruled out a diagnosis of Crohn's disease suspected by other modalities in three patients. CVE extended the involvement of Crohn's disease in three of the patients, and established new diagnoses in two patients. SUMMARY: CVE established new diagnoses, confirmed existing diagnoses, enlarged the extent of the disease, and ruled out the suspicion of Crohn's disease in 70% of the patients. Barium follow-through established diagnoses, measured the extent of the disease, and ruled out the suspicion of Crohn's disease in 37% of the patients. The capsule detected all of the lesions diagnosed by barium follow-through and entero-CT. CVE detected additional lesions that were not detected by other modalities in 47% of cases and ruled out lesions that were detected by other modalities in 16% of cases (P < 0.05). CONCLUSION: CVE is a superior and more sensitive procedure than barium follow-through and entero-CT in establishing the diagnosis and estimating the extent of Crohn's disease. PMID- 12655256 TI - Serum C-reactive protein correlates with survival in colorectal cancer patients but is not an independent prognostic indicator. AB - BACKGROUND: C-reactive protein (CRP) is an acute-phase protein synthesized in liver and up-regulated by pro-inflammatory cytokines, such as interleukin-6 (IL 6), interleukin-8 (IL-8) and tumour necrosis factor (TNF). Elevated CRP has been reported to be associated with reduced crude survival rates in patients with colorectal cancer. OBJECTIVE: To investigate the prognostic significance of preoperative serum CRP in relation to the disease-specific survival rate and expression of different cytokines. METHODS: One hundred and seventy-two consecutive patients with colorectal cancer, whose primary lesions were resected, were selected from April 1995 to December 1999. Preoperative serum CRP levels were measured, and the relationship between the elevation of CRP and clinicopathological factors was investigated. Prognostic significance was analysed by univariate and multivariate tests. RESULTS: One-third of patients had increased CRP levels, and this was associated with larger tumour size, lymph node or liver metastasis, and advanced Dukes' stage. Higher CRP levels were also related to the elevation of IL-6 and IL-8. The most important prognostic factor predicting survival was Dukes' stage (P < 0.001). Multivariate analysis indicated that CRP level is not an independent factor predicting survival. CONCLUSIONS: A preoperative elevation of serum CRP does not have the independent prognostic significance reported by earlier studies. Whether the elevated CRP can predict the development of cachexia or whether this association characterizes a pattern in tumour behaviour remains to be determined. PMID- 12655258 TI - Differentiation between patients with acute upper gastrointestinal bleeding who need early urgent upper gastrointestinal endoscopy and those who do not. A prospective study. AB - OBJECTIVE: To differentiate, among patients presented at the emergency department with acute upper gastrointestinal bleeding, those who need early urgent upper gastrointestinal endoscopy from those who do not. METHODS: Seventeen variables for each patient presenting with upper gastrointestinal bleeding were prospectively recorded and considered in a multivariate analysis. We used the presence of active bleeding during early urgent upper gastrointestinal endoscopy within 12 h from admission as the end point. The derived score was validated with data from the next consecutive patients presenting with upper gastrointestinal bleeding. RESULTS: Among 190 consecutive patients (mean age 63.7 +/- 16 years; 64.7% men), active bleeding was observed in 51 patients (26.8%). Four variables were identified as independent predictors (P < 0.05) of active bleeding in early urgent upper gastrointestinal endoscopy and were used for the derivation of the following integer-based scoring system: number of points = 6 (fresh blood in nasogastric tube) + 4 (haemodynamic instability) + 4 (haemoglobin < 8 g/dl) + 3 (white blood cell count > 12 000/microl). The validation study consisted of 110 patients (71 men; mean age 66.1 +/- 14 years; 28 patients [25.5%] with active bleeding). In this study, a cut off of < 7 points indicated absence of active bleeding and >/= 11 points indicated presence of active bleeding; this gave a sensitivity of 96%, specificity of 98%, positive predictive value of 96% and negative predictive value of 98%. CONCLUSIONS: Simple clinical and laboratory variables available at presentation can be used to differentiate patients with upper gastrointestinal bleeding who do not need an early urgent upper gastrointestinal endoscopy from those who do. PMID- 12655257 TI - Prognostic relevance of Fas (APO-1/CD95) ligand in human colorectal cancer. AB - OBJECTIVE: Fas ligand (FasL) is an important mediator of immune function and induces apoptosis by binding to its receptor Fas on sensitized cells. It has recently been shown that malignancies may express FasL and acquire immune privilege by inducing apoptosis of lymphocytes. Acquired resistance to Fas mediated apoptosis is known to be an early event in carcinogenesis. The aim of this study was to determine the extent of FasL expression in patients with colorectal cancer and examine its relationship with several prognostic pathological features and survival. DESIGN AND METHODS: Sixty-eight patients (median age 66 years) with colorectal cancer, whose diagnosis was made between 1988 and 1991 and in whom long-term follow-up was available, were evaluated. The tumours were of varying stages at diagnosis (eight Dukes' A, 28 Dukes' B, 23 Dukes' C and nine Dukes' D). The expression of FasL was detected immunohistochemically with a rabbit polyclonal IgG using the DAKO EnVision+ System. The specificity of FasL binding was confirmed by pre-incubation of the antibody with the immunizing peptide prior to staining. The relationship with several pathological features was determined using Kendall's tau-b correlation. Overall survival was estimated using the Kaplan-Meier product limit curves. Differences in observed survival were tested for statistical significance using the Mantel-Haenszel log rank test. Both the extent and intensity of staining were graded by a blinded observer. RESULTS: FasL was predominantly expressed in tumour epithelial cells in 88% of the cases. The positive staining of tumours varied in extent. FasL staining was higher in earlier Dukes' stage tumours in that the extent of FasL staining negatively correlated with Dukes' stage (Kendall tau-b = 0.22, P = 0.038). Consistent with this, the overall survival was better with a greater extent of FasL expression (log rank chi2 = 5.68, P = 0.017). There was a lower extent of FasL expression in mucinous adenocarcinomas (Kendall tau-b = 0.288, P = 0.01) and in those tumours with neural invasion (Kendall tau-b = 0.26, P = 0.03). No relationship was detected between FasL and tumour site, size, margin, differentiation, vascular invasion, necrosis or Crohn's-like reaction. CONCLUSIONS: FasL is widely expressed in colorectal cancers. This finding suggests that the extent of FasL expression in colorectal tumours is directly related to patients' survival. PMID- 12655259 TI - Mutant p53 expression and apoptotic activity of Helicobacter pylori positive and negative gastritis in correlation with the presence of intestinal metaplasia. AB - BACKGROUND: Mutation of the p53 gene is detectable in most cases of gastric cancer, as it is the most common genetic alteration in human malignancies. It is also well documented that Helicobacter pylori infection plays an important role in gastric carcinogenesis. There is still no clarification, however, concerning how genetic instability influences the homeostasis of gastric epithelium. We have studied the effect of H. pylori infection on apoptosis of the antral epithelium in the presence/absence of intestinal metaplasia and the expression of the p53 oncoprotein. The relationship between these two processes is analysed. METHODS: Antral biopsies were taken from 36 patients who underwent routine upper endoscopy (17 men, 19 women, mean age 61.0 years). The biopsies were fixed in formalin and embedded in paraffin. Patients were classified into two histological groups: (1) as chronic gastritis without intestinal metaplasia (n = 19), and (2) chronic gastritis with intestinal metaplasia (n = 17). An immunohistochemical method was used to detect the expression of p53 oncoprotein, and the terminal transferase mediated dUTP nick end-labelling (TUNEL) method was used to detect apoptotic cells. RESULTS: In the absence of intestinal metaplasia, both the apoptotic index (0.0272 +/- 0.011 vs 0.0128 +/- 0.006) and expresssion of p53 (35.55 +/- 31.16 vs 18.33 +/- 19.65) were significantly higher in H. pylori positive cases compared to H. pylori negative cases. In the presence of intestinal metaplasia, p53 expression was further increased (P < 0.05), but apoptosis was similar to that observed in H. pylori negative gastritis without intestinal metaplasia. In the presence of intestinal metaplasia, H. pylori infection did not influence apoptosis (0.013 +/- 0.004 vs 0.011 +/- 0.004), or p53 ratio (70.16 +/- 22.54 vs 68.50 +/- 28.96). In the sequence of gastritis-intestinal metaplasia the two indices show a close negative correlation (P < 0.05). CONCLUSION: In the absence of intestinal metaplasia H. pylori infection increases both apoptotic activity and expression of p53 oncoprotein in the gastric mucosa. The lack of increased apoptosis with a higher p53 expression in the presence of intestinal metaplasia suggests an increased genetic instability and also may suggest that mutation of the p53 gene is an early step in the multistep process of gastric carcinogenesis. PMID- 12655260 TI - How Helicobacter pylori urease may affect external pH and influence growth and motility in the mucus environment: evidence from in-vitro studies. AB - BACKGROUND: Survival of Helicobacter pylori is dependent upon urease in the cytoplasm and at the bacterial surface. We have sought to clarify how alkaline ammonium salts, released from urea by this enzyme, might alter mucus pH and so affect growth and motility of the bacterium in the gastric mucus environment. METHODS: Experiments were conducted in vitro to determine how the growth and motility of H. pylori are affected by changes in external pH, and how the bacterium, by hydrolysing urea, alters the pH of the bicarbonate buffer that occurs at the gastric mucosal surface. These data were fitted into experimental models that describe how pH varies within the mucus layer in the acid-secreting stomach. RESULTS: H. pylori was motile between pH 5 and 8, with optimal motility at pH 5. It grew between pH 6 and 8, with optimal growth at pH 6. The bacterium had urease activity between pH 2.7 and 7.4, as evidenced by pH rises in bicarbonate-buffered solutions of urea. Changes in buffer pH were dependent upon initial pH and urea concentration, with the greatest rate of pH change occurring at pH 3. Modelling experiments utilizing these data indicated that (1) in the absence of urease, H. pylori growth and motility in the mucus layer would be restricted severely by low mucus pH in the acid-secreting stomach, and (2) urease will sometimes inhibit H. pylori growth and motility in the mucus layer by elevating the pH of the mucus environment above pH 8. CONCLUSIONS: Urease is essential to the growth and motility of H. pylori in the mucus layer in the acid secreting stomach, but, paradoxically, sometimes it might suppress colonization by raising the mucus pH above 8. This latter effect may protect the bacteria from the adverse consequences of overpopulation. PMID- 12655261 TI - Effects of selective COX-2 inhibitors on the gastric permeability of sucrose: a controlled study with placebo and ibuprofen. AB - OBJECTIVE: Acute and chronic use of non-steroidal anti-inflammatory drugs can increase gastrointestinal permeability. Celecoxib, which selectively inhibits the enzyme cyclooxygenase-2, is a novel anti-inflammatory drug with minimal gastrointestinal toxic effects while retaining anti-inflammatory efficacy. Our aim was to assess the potential effects of celecoxib on gastric permeability in comparison with placebo and ibuprofen. DESIGN: We conducted a prospective, double blind, cross-over study. SETTING: This study is carried out at Marmara University Hospital. PARTICIPANTS: Twenty-five healthy subjects entered the study but 19 subjects completed the treatment. INTERVENTION: Subjects were randomized to celecoxib 100 mg twice daily, ibuprofen 600 mg twice daily or placebo for 7 days in pre-defined sequences. Treatments were separated by a 7 day washout period. MAIN OUTCOME MEASURE: Gastric permeability was assessed by measuring urinary excretion of sucrose spectrophotometrically. RESULTS: Ibuprofen 600 mg twice daily produced greater increases in gastric permeability compared with placebo or celecoxib (geometric mean of urinary sucrose recovery was 59.15, 32.65 and 33.11 mg/h for ibuprofen, placebo and celecoxib, respectively) (P < 0.001). Celecoxib was generally better tolerated than ibuprofen. CONCLUSIONS: When compared with ibuprofen, celecoxib 100 mg twice daily has no significant effect on gastric mucosa in healthy subjects. PMID- 12655262 TI - A primary care cross-sectional study of undiagnosed adult coeliac disease. AB - OBJECTIVES: To establish the prevalence of coeliac disease in the general population and in specific conditions, such as irritable bowel syndrome, iron deficiency anaemia, fatigue and other coeliac-related conditions. METHODS: Primary-care-based cross-sectional study using immunoglobulins, IgA/IgG antigliadin antibodies and endomysial antibodies to initially recognize coeliac disease. A total of 1200 volunteers were recruited from January 1999 to June 2001 from five general practices in South Yorkshire, UK. Any participant with a positive IgA antigliadin antibody, positive endomysial antibody, or only IgG antigliadin antibody in the presence of IgA deficiency was offered a small-bowel biopsy to confirm the diagnosis of coeliac disease. RESULTS: Twelve new cases of coeliac disease were diagnosed from 1200 samples. The prevalence of coeliac disease in this primary care population sample is 1% (95% CI 0.4-1.3%). The prevalence of coeliac disease was 3.3% (4/123) in participants with irritable bowel syndrome, 4.7% (3/64) in participants with iron deficiency anaemia, and 3.3% (3/92) in participants with fatigue. CONCLUSIONS: This study describes the prevalence of undiagnosed adult coeliac disease in primary care patients with irritable bowel syndrome, iron deficiency anaemia and fatigue. Underdiagnosis of coeliac disease is common in primary care. A case-finding approach would avoid delays in diagnosis and the associated morbidity or potential complications of coeliac disease. A low threshold for serological screening of patients with coeliac-associated symptoms or conditions would be an optimal strategy. PMID- 12655263 TI - Idiopathic cholangiopathy in a biliary cast syndrome necessitating liver transplantation following head trauma. AB - The development of total biliary casts is very unusual, especially in patients who have not undergone liver transplantation. The aetiology of these casts is uncertain but several factors are believed to play a role, including periods of fasting, haemolysis, cholangitis and recent surgery. Resultant bile stasis and/or gallbladder hypocontractility promote sludge and subsequent stone formation. Here we present the case of a previously well 66-year-old woman who developed a total biliary cast several weeks after being involved in a road traffic accident during which she sustained head injuries but no obvious liver insult. This cast was removed at laparotomy but the patient had resultant diffuse biliary tree abnormalities and persistent cholestasis and subsequently required a liver transplant. The possible aetiologies of biliary cast formation and subsequently cholangiopathy necessitating transplantation in this patient are described. PMID- 12655264 TI - Severe alcoholic hepatitis accompanied by Fournier's gangrene. AB - Fulminant necrotizing soft-tissue infection of the external genitalia and perineum (Fournier's gangrene) occurred in a patient with severe alcoholic hepatitis. By means of radical debridement and disinfection of the necrotizing tissue, use of broad-spectrum antibiotics and prednisolone, and other supportive measures, Fournier's gangrene and severe alcoholic hepatitis eventually subsided with broad skin defects in the waist and external genitalia. Later, the skin defects were successfully reconstructed with skin grafts. Although the route of bacterial intrusion could not be defined, Fournier's gangrene presumably developed in a background of impaired immunological defences, principally associated with habitual massive alcohol consumption and profound liver dysfunction. This present case highlights not only the underlying immunocompromised condition but also the need for concern regarding the urogenital and anorectal areas as potential foci of life-threatening infection in patients with liver dysfunction, especially when related to alcohol intake. PMID- 12655265 TI - Autoimmune hepatitis, cryoglobulinaemia and untreated coeliac disease: a case report. AB - The clinical presentation of adult coeliac disease is often uncharacteristic, with extraintestinal symptoms being the main findings. We report a 48-year-old woman who presented with type II, hepatitis-C-negative cryoglobulinaemia, elevated liver enzymes, and iron deficiency. Antinuclear antibodies were positive, and immunoglobulin G (IgG) levels were elevated. On liver biopsy, a diagnosis of type I autoimmune hepatitis with a possible autoimmune cholangitis overlap syndrome was made. Immunosuppressive treatment led to a normalization of transaminase levels and resolved the cryoglobulinaemic vasculitis. In addition, the patient exhibited low ferritin and iron levels, which led to the diagnosis of coeliac disease. Long-standing, untreated coeliac disease is recognized to be a trigger for autoimmune disorders and is known to be associated with other autoimmune diseases, but the association with autoimmune hepatitis or autoimmune cholangitis is reported rarely. We conclude that in patients with autoimmune liver disease and unspecific clinical signs, such as iron deficiency, coeliac disease must be ruled out. PMID- 12655266 TI - Magnetic resonance imaging and magnetic resonance cholangiopancreaticography for diagnosis and pre-interventional evaluation of a fluid thoracic mass. AB - Differential diagnoses of various types of thoracic cystic masses are a prerequisite for appropriate management strategies. Mediastinal pancreatic pseudocysts are rare but life-threatening complications of pancreatitis. They may appear as a thoracic mass on X-rays and need to be differentiated from other lesions. This report demonstrates that magnetic resonance cholangiopancreaticography, which is not included in current algorithms, is a valuable diagnostic tool in such cases. We summarize the advantages and disadvantages of magnetic resonance cholangiopancreaticography and show how pre interventional identification of pancreatic origin and exclusion of necrotic debris guided the therapeutic management of the patient. However, after 3 weeks of successful percutaneous drainage, infectious complications necessitated surgical intervention. PMID- 12655267 TI - Decompensation of chronic stable alcoholic liver disease by severe exfoliative dermatitis. AB - Of the numerous precipitants of hepatic decompensation in chronic liver disease, there are no reports in the literature documenting an acute decompensation following an acute episode of severe dermatitis. This case highlights the haemodynamic consequences of a severe flare up of exfoliative dermatitis in a patient with stable chronic alcoholic liver disease, speculates on the mechanism by which this may provoke clinical decompensation and the impact this may have upon liver failure. PMID- 12655268 TI - Spontaneous, self-limited, non-atherosclerotic dissection of the superior mesenteric artery. AB - A case of spontaneous, non-atherosclerotic dissection of the superior mesenteric artery is described. The patient presented with severe abdominal pain. An initial computed tomography scan did not disclose any pathology, namely the major mesenteric arteries were perfused. The patient's initial symptoms resolved during the first 48 h under conservative treatment. Sagittal reconstruction from a follow-up computed tomography scan disclosed a 7 cm long dissection of the proximal superior mesenteric artery with thrombosis of the false lumen and residual blood flow through the true lumen. There were no signs of transmural infarction of the bowel. Anticoagulation with heparin and later with phenprocoumon was initiated and the patient is well after 6 months. Together with several other cases reported recently, and in contrast to previous experience, this case adds evidence that conservative treatment of spontaneous, non atherosclerotic dissection of the superior mesenteric artery may be an alternative to surgery if residual blood flow is maintained. PMID- 12655270 TI - Is biliary atresia an autoimmune disease? PMID- 12655269 TI - An unusual expression of hyperplastic gastropathy (Menetrier type) in twins. AB - Menetrier's disease is an uncommon condition of unknown aetiology. We describe two cases of male identical twins with haematemesis aged 29 and 35 years that exhibited a similar and particular form of this hyperplastic gastropathy. Their stomachs showed confluent polypoid mucosal projections affecting mainly the gastric fundus and the antrum. To the best of our knowledge, only four previous cases have been reported in a familial setting, and this is the first documented example of an occurrence in twins. These two cases suggest the possibility of a genetic predisposition for this condition. PMID- 12655271 TI - Genotype distribution of hepatitis C virus in the Hungarian population with chronic viral hepatitis C. PMID- 12655273 TI - Mechanisms of free radical production in the vascular wall. PMID- 12655274 TI - Oxidative stress and heart failure. PMID- 12655275 TI - Oxidative stress and regional ischemia-reperfusion injury: the peroxynitrite poly(ADP-ribose) polymerase connection. PMID- 12655276 TI - Redox regulation following cardiac ischemia and reperfusion. PMID- 12655277 TI - The conceptual basis of mathematics in cardiology: (II). Calculus and differential equations. AB - This is the second in a series of four articles developed for the readers of Coronary Artery Disease. Without language ideas cannot be articulated. What may not be so immediately obvious is that they cannot be formulated either. One of the essential languages of cardiology is mathematics. Unfortunately, medical education does not emphasize, and in fact, often neglects empowering physicians to think mathematically. Reference to statistics, conditional probability, multicompartmental modeling, algebra, calculus and transforms is common but often without provision of genuine conceptual understanding. At the University of Vermont College of Medicine, Professor Bates developed a course designed to address these deficiencies. The course covered mathematical principles pertinent to clinical cardiovascular and pulmonary medicine and research. It focused on fundamental concepts to facilitate formulation and grasp of ideas. This series of four articles was developed to make the material available for a wider audience. The articles will be published sequentially in Coronary Artery Disease. Beginning with fundamental axioms and basic algebraic manipulations they address algebra, function and graph theory, real and complex numbers, calculus and differential equations, mathematical modeling, linear system theory and integral transforms and statistical theory. The principles and concepts they address provide the foundation needed for in-depth study of any of these topics. Perhaps of even more importance, they should empower cardiologists and cardiovascular researchers to utilize the language of mathematics in assessing the phenomena of immediate pertinence to diagnosis, pathophysiology and therapeutics. The presentations are interposed with queries (by Coronary Artery Disease abbreviated as CAD) simulating the nature of interactions that occurred during the course itself. Each article concludes with one or more examples illustrating application of the concepts covered to cardiovascular medicine and biology. PMID- 12655278 TI - Glutathione peroxidase 1 genotype is associated with an increased risk of coronary artery disease. AB - OBJECTIVE: Oxidative stress is implicated in the pathogenesis of many human diseases including atherosclerosis. Human glutathione peroxidase 1 (hgpx1) participates in limiting cellular damage caused by oxidation. A characteristic polyalanine sequence polymorphism in exon 1 of hgpx1 produces three alleles with five, six or seven alanine (ALA) repeats in this sequence. The objective of this study was to determine whether hgpx1 genotype is associated with an altered risk of coronary artery disease (CAD). METHODS: The frequency of the ALA6 allele was determined in 207 men with angiographic evidence of significant CAD compared to a control group (n = 146), by analysing the lengths of polymerase chain reaction fragments containing the ALA repeat polymorphism. Additional information was collected on severity of CAD, presence or absence of a prior acute myocardial infarction (AMI), smoking status, body mass index (BMI) and other clinical data. RESULTS: There was a significant association between individuals with at least one ALA6 allele and an increased risk of CAD after adjustment for age, BMI and smoking status (odds ratio, 2.07, 95% confidence interval, 1.08-3.99, P = 0.029). However, there was no association between hgpx1 genotype and a previous history of AMI or hgpx1 genotype and severity of CAD. CONCLUSION: We conclude that individuals possessing one or two ALA6 alleles appear to be at a modest increased risk of CAD. This observation merits further investigation in other patient populations. PMID- 12655279 TI - Vascular endothelial function in patients with slow coronary flow. AB - BACKGROUND: Slow coronary flow (SCF) in a normal coronary angiogram is a well recognized clinical entity, but its etiopathogenesis remains unclear. DESIGN: The aim of the study was to determine endothelial function in patients with SCF using a flow-mediated dilatation (FMD) technique in the brachial artery. METHODS: Coronary flow was quantified using the corrected thrombosis in myocardial infarction (TIMI) frame count (CTFC) method. Endothelial function was studied in 27 patients with SCF (23 men, four women, mean age 47.6+/-8.7 years) and in 30 people with normal coronary flow (NCF) (22 men and eight women, mean age 47.5+/ 7.4 years). RESULTS: The flow-mediated diameter increase in the SCF group was significantly smaller than that in the NCF group (3.48+/-0.10% compared with 9.11+/-0.10%, P < 0.001). The percentage of nitroglycerine (NTG)-induced dilatation was not significantly different between patients with SCF and people with NCF (16.8+/-1.1% compared with 17.1+/-1.1%, P = 0.87). Simple regression analysis showed that mean CTFC (CTFC(m)) was strongly and inversely related to the percentage of FMD (r = -0.29, P < 0.01) in all participants. When the patients with SCF were excluded, CTFC(m) was still inversely related to the percentage of FMD (r = -0.36, P < 0.05). CTFC(m) was also inversely related to NTG-induced dilatation in the 57 participants (r = -0.23, P < 0.05). Multiple regression analysis showed that CTFC(m) was inversely related to the percentage of FMD only (r = -0.37, P < 0.05). CONCLUSIONS: These findings suggest that endothelial function is impaired in people with SCF and that CTFC correlates well with endothelial dysfunction. PMID- 12655281 TI - Efficacy and tolerability of trimetazidine in stable angina: a meta-analysis of randomized, double-blind, controlled trials. AB - OBJECTIVE: The objective of this meta-analysis was to evaluate the efficacy and tolerance of the metabolic agent trimetazidine (TMZ), both in monotherapy and in combination with other antianginal agents, in the treatment of stable angina pectoris. A search of literature published between 1985 and 2001 was performed on computerized databases (MEDLINE and EMBASE). METHODS: Only double-blind, randomized, controlled trials were included in this meta-analysis. Patients had to be treated for at least 2 weeks. Four parameters were selected, one clinical parameter (number of weekly angina attacks) and three ergometric parameters (time to 1 mm ST-segment depression, total work and exercise duration at peak exercise). They were evaluated at baseline and at the end of the treatment period.The quality of the trials was assessed on specific methodological criteria. Standard statistical methods, pooled odds ratio and 95% confidence intervals for subjective symptoms and pooled z and P for objective symptoms, were used. RESULTS: Twelve clinical studies meeting our criteria were analyzed. Results showed that TMZ significantly reduced the number of weekly angina attacks in coronary patients and improved time to 1 mm segment depression and total work at peak exercise, while exercise duration at peak exercise showed a trend toward improvement (P = 0.09). CONCLUSION: This meta-analysis confirms the efficacy of TMZ in the treatment of stable angina, compared with placebo or conventional antianginal agent, as well as in monotherapy or in combination with conventional antianginal agents. TMZ is well tolerated in monotherapy as well as in combination. PMID- 12655280 TI - Antibodies to oxidized low-density lipoprotein in patients following coronary artery revascularization. AB - BACKGROUND: Measurement of antibodies to oxidized low-density lipoprotein (AB oxLDL) has been proposed as a way of identifying individuals at risk for cardiovascular disease beyond the determination of LDL cholesterol (LDL-c) alone. While this hypothesis is appealing, evidence that AB-oxLDL is a risk factor independent of LDL-c levels remains to be established. METHODS AND RESULTS: We conducted a cross-sectional study of 158 hypercholesterolemic adults who underwent coronary revascularization. Patients were randomized in a clinical trial to a nurse case management (NURS) or an enhanced usual care (EUC) group. The goal in the NURS group was to achieve an LDL-c <85 mg/dl with pharmacotherapy and lifestyle advice. Six months after revascularization, serum and urine were collected to determine lipids, AB-oxLDL, urinary isoprostanes (an in-vivo marker of oxidative damage), oxygen-radical-absorbing capacity (ORAC) of serum, serum carotenoids, alpha-tocopherol and C-reactive protein (CRP). After 6 months of intervention, the NURS group had a mean (+/-SD) LDL-c of 84 (18) mg/dl compared with 105 (25) mg/dl in the EUC group (P < 0.001). In addition, AB-oxLDL was lower (median +/- 95% confidence interval, -202 milliunits (mU)/ml, -372 to -32, P = 0.02) in the NURS group (488 mU/ml, 399, 588) than in the EUC group (690 mU/ml, 544, 847). The difference in AB-oxLDL between groups persisted after adjustment for LDL-c (-316 mU/ml, -519 to -112, P = 0.005). Further adjustment for ORAC and alpha-tocopherol did not diminish the group effect. The groups did not differ significantly in serum antioxidants, urinary isoprostane excretion or CRP. In additional analyses, change in LDL-c from baseline was not associated with AB oxLDL at 6 months. CONCLUSIONS: Our results demonstrate lower AB-oxLDL in patients who achieve greater lipid reduction. However, AB-oxLDL appears to be independent of achieved LDL-c, ORAC levels or serum antioxidant levels. These results support the hypothesis that AB-oxLDL is independent of LDL-c level. Further prospective studies are needed to determine the prognostic value of AB oxLDL. PMID- 12655282 TI - Evaluation of the Oculomedin gene in the etiology of primary open angle and exfoliative glaucoma. AB - PURPOSE: Glaucoma is a disease of the retinal ganglion cells leading to reduction of peripheral vision. It is often associated with an increase in intraocular pressure, leading to mechanical stress of tissues. The oculomedin gene is activated by such stretching and is therefore a candidate for causing glaucoma. METHODS: The coding sequence and part of the promoter was screened for sequence variants in Swedish cohorts of primary open angle glaucoma, exfoliative glaucoma, and matched controls. RESULTS: Only rare variants were detected in the patient material. CONCLUSIONS: There was no evidence that the oculomedin gene participates in the etiology of glaucoma. PMID- 12655283 TI - Effects of the matricellular protein SPARC on human retinal pigment epithelial cell behavior. AB - PURPOSE: To determine the effects of the matricellular protein SPARC (Secreted Protein, Acidic and Rich in Cysteine) on human retinal pigment epithelial (HRPE) cell behavior in vitro. METHODS: Proliferation and migration assays were performed on HRPE cells exposed to various concentrations of SPARC. Additionally, HRPE cells were seeded on top of collagen matrices (a 2D model of the retinal scarring disorder known as proliferative vitreoretinopathy or PVR) and were exposed to SPARC over a 7 day period. Changes in matrix contraction were recorded. RESULTS: HRPE cell proliferation was significantly inhibited at 1 and 10 microg/ml SPARC (p<0.01). SPARC protein did not stimulate HRPE cell migration at any of the concentrations used. SPARC did not significantly affect fibronectin induced HRPE cell migration at SPARC concentrations up to 10 microg/ml. HRPE cell seeded collagen matrices demonstrated a significant inhibition of matrix contraction by 1 and 10 microg/ml SPARC (t-test; p<0.02 and 0.001, respectively) compared to controls. CONCLUSIONS: SPARC protein has anti-proliferative effects on HRPE cells in vitro. In addition, SPARC appears to have an inhibitory effect on HRPE-mediated contraction of 2D collagen matrices. These results are consistent with an important role for SPARC in modulating cell behavior in vitro and may indicate a role for SPARC in modifying HRPE cell activities during the development of PVR and other proliferative retinal diseases. PMID- 12655285 TI - Keratocyte matrix interactions and thrombospondin 2. AB - PURPOSE: To determine whether human keratocytes synthesize thrombospondins 2 and 3 (TSP-2, TSP-3) in a collagen matrix and the effect of addition of antibodies to TSP-2 and TSP-3 on keratocyte-populated collagen matrix contraction. METHODS: Keratocyte-populated collagen matrices were evaluated for TSP-2 and TSP-3 mRNA. Sections of matrices were stained by immunohistochemistry for TSP-2 and TSP-3. Keratocyte populated collagen matrices were treated with antibodies specific for TSP-2 and TSP-3 and these preparations were evaluated for contraction and keratocyte morphology. RESULTS: Keratocyte derived fibroblasts in collagen matrices contained TSP-2 and TSP-3 mRNA, and the cells were immunoreactive for both proteins. Compared to controls, an antibody specific to the N-terminal domain of TSP-2 significantly inhibited matrix contraction for up to 10 days at a concentration of 20 microg/ml antibody. At 2 microg/ml TSP-2 antibody concentration significant inhibition occurred up to 3 days. Removal of the antibody from the media reversed the inhibitory effect. Cultured keratocytes in TSP-2 treated collagen matrices appeared more rounded than keratocytes in control matrices. An antibody specific to TSP-3 had no effect on matrix contraction or keratocyte morphology. CONCLUSIONS: Keratocyte derived fibroblasts synthesize TSP 2 and TSP-3 when seeded in collagen matrices. Antibody specific to TSP-2 reversibly inhibits matrix contraction. TSP-2 may play a key role in keratocyte/collagen matrix interactions, as may occur during corneal stromal repair. PMID- 12655284 TI - Expression of cone photoreceptor cGMP-phosphodiesterase alpha' subunit in Chinese hamster ovary, 293 human embryonic kidney, and Y79 retinoblastoma cells. AB - PURPOSE: A functional protein is required for structure/function analysis of cone photoreceptor cGMP-phosphodiesterase alpha' subunit (PDEalpha'). The purpose of this study was to express enzymatically active PDEalpha'. METHODS: Three expression vectors were constructed for transient and stable expression of PDEalpha': pC57 (transient) was obtained by subcloning bovine PDEalpha' cDNA into the pCIS2 expression vector; pNC57 (stable) was constructed by inserting the neo gene controlled by the mouse phosphoglycerate kinase-1 gene promoter into the pC57 vector; and pFC57 (transient) was generated by fusing the sequence encoding the FLAG peptide to the 5' end of the coding region of PDEalpha' cDNA. The recombinant plasmid DNAs were introduced into HEK293, CHO, or Y79 retinoblastoma cells using the calcium phosphate-mediated transfection procedure or lipofectamin. Northern and western blot hybridizations were used for RNA and protein analysis, respectively. RESULTS: Northern blots of both HEK293- and CHO transfected cells showed strong expression of a 3 kb transcript corresponding to PDEalpha'. cGMP-PDE activity measured in homogenates of transiently and stably transfected cells ranged between 1.5 and 2.2 nmol cGMP hydrolyzed/min x mg total protein, a level of PDE activity slightly greater than that previously reported for the individual rod-photoreceptor PDE subunits transiently-expressed in HEK293 cells. Western blots of these cell homogenates showed a low level of expressed PDEalpha'. Transfection of Y79 retinoblastoma cells, that have been shown to express rod and cone PDEs endogenously, with the construct containing cone PDEa' cDNA fused to the FLAG peptide resulted in a protein with no enzymatic activity. CONCLUSIONS: Our results demonstrate that both HEK293 and CHO cells are capable of expressing functionally active cone PDEalpha'. High level of mRNA transcription and relatively low protein synthesis efficiency indicates the presence of a post-transcriptional control mechanism regulating overall expression of PDEalpha' in HEK293 and CHO cells. PMID- 12655287 TI - Death receptors leave a caspase footprint that Smacs of XIAP. PMID- 12655288 TI - Role of membrane microdomain rafts in TNFR-mediated signal transduction. PMID- 12655289 TI - Death fold domain interaction in apoptosis. PMID- 12655290 TI - JNK: a killer on a transcriptional leash. PMID- 12655291 TI - Apaf-1/cytochrome c apoptosome: an essential initiator of caspase activation or just a sideshow? PMID- 12655292 TI - All in the family: evolutionary and functional relationships among death receptors. AB - Over the last decade, significant progress has been made towards identifying the signaling pathways within mammalian cells that lead to apoptosis mediated by death receptors. The simultaneous expression of more than one death receptor in many, if not all, cell types suggests that functional innovation has driven the divergence of these receptors and their cognate ligands. To better understand the physiological divergence of the death receptors, a phylogenetic analysis of vertebrate death receptors was conducted based upon amino-acid sequences encoding the death domain regions of currently known and newly identified members of the family. Evidence is presented to indicate an ancient radiation of death receptors that predates the emergence of vertebrates, as well as ongoing divergence of additional receptors both within several receptor lineages as well as modern taxonomic lineages. We speculate that divergence among death receptors has led to their functional specialization. For instance, some receptors appear to be primarily involved in mediating the immune response, while others play critical roles during development and tissue differentiation. The following represents an evolutionary approach towards an understanding of the complex relationship among death receptors and their proposed physiological functions in vertebrate species. PMID- 12655293 TI - The CD95(APO-1/Fas) DISC and beyond. AB - CD95 (APO-1/Fas) is a prototype death receptor characterized by the presence of an 80 amino acid death domain in its cytoplasmic tail. This domain is essential for the recruitment of a number of signaling components upon activation by either agonistic anti-CD95 antibodies or cognate CD95 ligand that initiate apoptosis. The complex of proteins that forms upon triggering of CD95 is called the death inducting signaling complex (DISC). The DISC consists of an adaptor protein and initiator caspases and is essential for induction of apoptosis. A number of proteins have been reported to regulate formation or activity of the DISC. This review discusses recent developments in this area of death receptor research. PMID- 12655294 TI - Signaling and transcriptional control of Fas ligand gene expression. AB - Fas ligand (FasL), a member of the tumor necrosis factor family, initiates apoptosis by binding to its surface receptor Fas. As a consequence, there is sequential activation of caspases and the release of cytochrome c from the mitochondria, with additional caspase activation followed by cellular degradation and death. Recent studies have shed important insight into the molecular mechanisms controlling FasL gene expression at the level of transcription. Nuclear factors such as nuclear factor in activated T cells, nuclear factor-kappa B, specificity protein-1, early growth response factor, interferon regulatory factor, c-Myc and the forkhead transcriptional regulator, alone or cooperatively, activate FasL expression. These factors are often coexpressed with FasL in pathophysiologic settings including human atherosclerotic lesions. Here, we review these important advances in our understanding of the signaling and transcriptional mechanisms controlling FasL gene expression. PMID- 12655295 TI - Tumor necrosis factor signaling. AB - A single mouse click on the topic tumor necrosis factor (TNF) in PubMed reveals about 50,000 articles providing one or the other information about this pleiotropic cytokine or its relatives. This demonstrates the enormous scientific and clinical interest in elucidating the biology of a molecule (or rather a large family of molecules), which began now almost 30 years ago with the description of a cytokine able to exert antitumoral effects in mouse models. Although our understanding of the multiple functions of TNF in vivo and of the respective underlying mechanisms at a cellular and molecular level has made enormous progress since then, new aspects are steadily uncovered and it appears that still much needs to be learned before we can conclude that we have a full comprehension of TNF biology. This review shortly covers some general aspects of this fascinating molecule and then concentrates on the molecular mechanisms of TNF signal transduction. In particular, the multiple facets of crosstalk between the various signalling pathways engaged by TNF will be addressed. PMID- 12655296 TI - Apo2L/TRAIL and its death and decoy receptors. AB - Apo2 ligand or tumour necrosis factor-related apoptosis-inducing ligand (Apo2L/TRAIL) is one of the several members of the tumour necrosis factor (TNF) gene superfamily that induce apoptosis through engagement of death receptors (DRs). Apo2L/TRAIL interacts with an unusually complex receptor system of two DRs and three decoys. This protein has garnered intense interest as a potential candidate for cancer therapy because as a trimer it selectively induces apoptosis in many transformed cells but not in normal cells. While much of the early characterisation of Apo2L/TRAIL and its receptors relied on overexpression studies, recent work using untransfected cells has clarified how endogenous proteins transmit apoptotic signals from this ligand. In this review, we focus on the apoptotic signalling pathways stimulated by Apo2L/TRAIL and summarise what is known about its physiological role. PMID- 12655297 TI - Many cuts to ruin: a comprehensive update of caspase substrates. AB - Apoptotic cell death is executed by the caspase-mediated cleavage of various vital proteins. Elucidating the consequences of this endoproteolytic cleavage is crucial for our understanding of cell death and other biological processes. Many caspase substrates are just cleaved as bystanders, because they happen to contain a caspase cleavage site in their sequence. Several targets, however, have a discrete function in propagation of the cell death process. Many structural and regulatory proteins are inactivated by caspases, while other substrates can be activated. In most cases, the consequences of this gain-of-function are poorly understood. Caspase substrates can regulate the key morphological changes in apoptosis. Several caspase substrates also act as transducers and amplifiers that determine the apoptotic threshold and cell fate. This review summarizes the known caspase substrates comprising a bewildering list of more than 280 different proteins. We highlight some recent aspects inferred by the cleavage of certain proteins in apoptosis. We also discuss emerging themes of caspase cleavage in other forms of cell death and, in particular, in apparently unrelated processes, such as cell cycle regulation and cellular differentiation. PMID- 12655298 TI - Caspase-2 redux. AB - It has been difficult to assign caspase-2 to the effector or initiator caspase groups. It bears sequence homology to initiators (caspase-9 and CED-3), but its cleavage specificity is closer to the effectors (caspase-3 and -7). Interest in caspase-2 was dampened by the lack of a dramatic phenotype in the caspase-2 null mouse. Studies have been inhibited by the lack of knowledge about its mechanism of activation and the lack of specific methods to assay its activity. Molecular studies have defined a unique role for caspase-2 in apoptosis initiated by beta amyloid toxicity or by trophic factor deprivation. Recently, a role for caspase-2 as an upstream initiator of mitochondrial permeabilization has been proposed. Thus, while much remains to be deciphered about caspase-2, most critically the mode of activation, it is clear that caspase-2 plays critical and singular roles in the control of programmed cell death. PMID- 12655299 TI - Degradation of chromosomal DNA during apoptosis. AB - Apoptosis is often accompanied by degradation of chromosomal DNA. CAD, caspase activated DNase, was identified in 1998 as a DNase that is responsible for this process. In the last several years, mice deficient in the CAD system have been generated. Studies with these mice indicated that apoptotic DNA degradation occurs in two different systems. In one, the DNA fragmentation is carried out by CAD in the dying cells and in the other, by lysosomal DNase II after the dying cells are phagocytosed. Several other endonucleases have also been suggested as candidate effectors for the apoptotic degradation of chromosomal DNA. In this review, we will discuss the mechanism and role of DNA degradation during apoptosis. PMID- 12655300 TI - Protein-based therapeutic approaches targeting death receptors. AB - Death receptors (DRs) are a growing family of transmembrane proteins that can detect the presence of specific extracellular death signals and rapidly trigger cellular destruction by apoptosis. Eight human DRs (Fas, TNF-R1, TRAMP, TRAIL-R1, TRAIL-R2, DR-6, EDA-R and NGF-R) have been identified. The best studied to date is Fas (CD95). Expression and signaling by Fas and its ligand (FasL, CD95L) is a tightly regulated process essential for key physiological functions in a variety of organs, including the maintenance of immune homeostasis. Recently, strong evidence has shown that dysregulation of Fas expression and/or signaling contributes to the pathogenesis of tissue destructive diseases such as graft versus-host disease, toxic epidermal necrolysis, multiple sclerosis and stroke. With these new developments, strategies for modulating the function of Fas signaling have emerged and provided novel protein-based therapeutic possibilities that will be discussed herein. Selective triggering of DR-mediated apoptosis in cancer cells is an emerging approach that is being intensely investigated as a mode of cancer therapy. Local administration of Fas agonists, and more promisingly, systemic use of soluble recombinant forms of TRAIL have shown efficacy in preclinical models of the disease. Developments in this field that may have important clinical implications for the treatment of cancer are reviewed. PMID- 12655301 TI - Autoimmune lymphoproliferative syndromes: genetic defects of apoptosis pathways. AB - Human and mouse natural mutants presenting with lymphoproliferative syndrome and autoimmunity (ALPS) have enlightened the role of the Fas and FasL in lymphocyte cell death and peripheral tolerance. Further study of the genetic basis of the human pathology led to the identification of apoptosis signaling defect, and pointed out to the crucial role of caspase-10 in the process of apoptosis induction. In contrast, the absence of lymproliferation in engineered mutants of 'death pathways' suggests that additional events are necessary to recapitulate the overt phenotype of ALPS patients or MRL/lpr mice. Moreover, these models highlight the roles of Fas and associated molecules, such as FADD and caspase-8, in lymphocyte development or activation. This review will discuss the main findings provided by the study of mouse models and human conditions. PMID- 12655303 TI - Mutually regulated expression of caspase-activated DNase and its inhibitor for apoptotic DNA fragmentation. PMID- 12655302 TI - Neutralization of TRAIL death pathway protects human neuronal cell line from beta amyloid toxicity. AB - Here we report that a novel member of the TNF-alpha family, TNF-related apoptosis inducing ligand (TRAIL), contributes substantially to amyloid-induced neurotoxicity in human SH-SY5Y neuronal cell line. Involvement of TRAIL in the amyloid-induced cell death is supported by cDNA array, Northern blot, and Western blot data, demonstrating increased TRAIL expression after treatment of the cells with a neurotoxic fragment of amyloid protein (betaAP). TRAIL was also found to be released in the culture media after betaAP treatment with a time-course overlapping to contents of the intracellular protein. Contribution of TRAIL to betaAP neurotoxicity is demonstrated by data showing that TRAIL-neutralizing monoclonal antibody protects neuronal SH-SY5Y cells from betaAP neurotoxicity. Moreover, exposure of neuronal SH-SY5Y cells to TRAIL leads to cell death, indicating that this substance per se is endowed with neurotoxic properties. We also found that, similarly to betaAP and TRAIL, activation of the death-domain adaptor protein FADD results in neuronal cell death. Lack of FADD function, by overexpression of its dominant negative, rescued cells from either TRAIL- or betaAP-induced neurotoxicity, supporting the hypothesis that these three molecules share common intracellular pathways. Finally, we found that betaAP strongly activated caspase-8, and the cell-permeable, selective caspase-8 inhibitor z-IETD-FMK prevents both betaAP- and TRAIL-induced neurotoxicity. In view of TRAIL's potency in inducing neuronal death, and its role as mediator of betaAP, it is plausible to hypothesize that TRAIL can be regarded as a molecule that provides substantial contribution to betaAP-dependent cell death, which takes part in the progression of the neurodegenerative process and related chronic inflammatory response. PMID- 12655304 TI - The active caspase-8 heterotetramer is formed at the CD95 DISC. PMID- 12655305 TI - Serotonin transporter: a potential substrate in the biology of suicide. AB - Suicide is a serious public health problem in the US, yet its neurobiological underpinnings are poorly understood. Suicide is highly correlated with depressive symptoms, and considerable evidence suggests that depression is associated with a relative deficiency in serotonergic neurotransmission. Serotonergic circuits also mediate impulsivity, a trait obviously relevant to suicide. These findings, taken together, suggest that alterations in the serotonergic system might contribute to suicidal behavior, serving as an impetus for researchers to scrutinize the serotonin transporter (SERT) as a potential substrate for the pathophysiology of suicide. Using post-mortem brain tissue, platelets, and DNA from suicide completers and attempters have not provided unequivocal evidence for a pre eminent role for the SERT in the pathophysiology of suicide. This paper provides a review of several studies that have evaluated the role of the SERT in the pathophysiology of suicide. PMID- 12655306 TI - Sex-dependent modulation of ethanol consumption in vesicular monoamine transporter 2 (VMAT2) and dopamine transporter (DAT) knockout mice. AB - Several lines of evidence suggest that monoaminergic systems, especially dopaminergic and serotoninergic systems, modulate ethanol consumption. Humans display significant differences in expression of the vesicular and plasma membrane monoamine transporters important for monoaminergic functions, including the vesicular monoamine transporter (VMAT2, SLC18A2) and dopamine transporter (DAT, SLC6A3). In addition, many ethanol effects differ by sex in both humans and animal models. Therefore, ethanol consumption and preference were compared in male and female wild-type mice, and knockout (KO) mice with deletions of genes for DAT and VMAT2. Voluntary ethanol (2-32% v/v) and water consumption were compared in two-bottle preference tests in wild-type (+/+) vs heterozygous VMAT2 KO mice (+/-) and in wild-type (+/+) vs heterozygous (+/-) or homozygous (-/-) DAT KO mice. Deletions of either the DAT or VMAT2 genes increased ethanol consumption in male KO mice, although these effects were highly dependent on ethanol concentration, while female DAT KO mice had higher ethanol preferences. Thus, lifetime reductions in the expression of either DAT or VMAT2 increase ethanol consumption, dependent on sex. PMID- 12655307 TI - Previous exposure to VTA amphetamine enhances cocaine self-administration under a progressive ratio schedule in an NMDA, AMPA/kainate, and metabotropic glutamate receptor-dependent manner. AB - Previous exposure to amphetamine (AMPH) in the ventral tegmental area (VTA) enhances cocaine self-administration in a D(1) dopamine receptor-dependent manner. The present study examined the contribution of VTA NMDA, AMPA/kainate, and metabotropic glutamate (mGlu) receptors to this effect. Rats in different groups received three intra-VTA injections, one every third day, of either saline (0.5 microl/side), AMPH (2.5 microg/0.5 microl/side), AMPH+CPP (NMDA receptor antagonist; 10 microM or 100 microM/0.5 microl/side), AMPH+CNQX (AMPA/kainate receptor antagonist; 0.3 mM or 1 mM/0.5 microl/side), AMPH+MCPG (mGlu receptor antagonist; 0.5 mM or 50 mM/0.5 microl/side), or the glutamate receptor antagonists alone. Starting 7-10 days after the last pre-exposure injection, rats were trained to self-administer cocaine (0.3 mg/kg/infusion) and then tested under a progressive ratio (PR) schedule of reinforcement for 6 consecutive days. As reported previously, VTA AMPH pre-exposed rats worked more and obtained more infusions of cocaine than saline pre-exposed animals. Coadministration of CPP, CNQX, or MCPG with AMPH during pre-exposure dose-dependently blocked this enhancement of cocaine self-administration. Rats pre-exposed to the glutamate receptor antagonists alone did not differ on the test days from the saline pre exposed controls. These results indicate that, in a manner paralleling the induction of sensitization of the locomotor stimulating effects of AMPH, activation of NMDA, AMPA/kainate, and mGlu receptors during pre-exposure to AMPH in the VTA is necessary for the enhancement of cocaine self-administration to develop. PMID- 12655308 TI - Amphetamine effects on prepulse inhibition across-species: replication and parametric extension. AB - Despite the similarities of prepulse inhibition (PPI) of the startle reflex and its apparent neural regulation in rodents and humans, it has been difficult to demonstrate cross-species homology in the sensitivity of PPI to pharmacologic challenges. PPI is disrupted in rats by the indirect dopamine (DA) agonist amphetamine, and while studies in humans have suggested similar effects of amphetamine, these effects have been limited to populations characterized by smoking status and specific personality features. In the context of a study assessing the time course of several DA agonist effects on physiological variables, we failed to detect PPI-disruptive effects of amphetamine in a small group of normal males. The present study was designed to reexamine this issue, using a larger sample and a paradigm that should be more sensitive for detecting drug effects. PPI in rats was shown to be disrupted by the highest dose of amphetamine (3.0 mg/kg) at relatively longer prepulse intervals (>30 ms). In humans, between-subject comparisons of placebo (n=15) vs 20 mg amphetamine (n=15) failed to detect significant PPI-disruptive effects of amphetamine, but significant PPI-disruptive effects at short (10-20 ms) prepulse intervals were detected using within-subject analyses of postdrug PPI levels relative to each subject's baseline PPI. Post hoc comparisons failed to detect greater sensitivity to amphetamine among subjects characterized by different personality and physiological traits. Bioactivity of amphetamine was verified by autonomic and subjective changes. These results provide modest support for cross-species homology in the PPI-disruptive effects of amphetamine, but suggest that these effects in humans at the present dose are subtle and may be best detected using within-subject designs and specific stimulus characteristics. PMID- 12655309 TI - A systemically administered neurotensin agonist blocks disruption of prepulse inhibition produced by a serotonin-2A agonist. AB - Prepulse inhibition (PPI) of the startle reflex can be disrupted by drugs that act as agonists at the serotonin (5-HT) 2A receptor, such as DOI, and this effect is blocked by drugs that inhibit 5-HT2A transmission. We tested the effects of systemic administration of PD149163, a neurotensin agonist, on DOI-induced disruption of PPI in Sprague-Dawley rats. PD149163 completely and dose dependently blocked the PPI deficits produced by DOI. These findings suggest that, in addition to their established ability to inhibit dopamine transmission, neurotensin agonists may also inhibit 5-HT2A transmission, a pharmacological feature associated with atypical antipsychotic drugs. PMID- 12655310 TI - GABA-A and 5-HT1A receptor agonists block expression of fear-potentiated startle in mice. AB - The present experiments characterized the acquisition of fear-potentiated startle (FPS) and determined the sensitivity of FPS to anxiolytic compounds in DBA/1J mice. A light (30 s) conditioned stimulus (CS) and mild footshock (0.14 mA, 0.5 s) unconditioned stimulus (US) were used. First, acquisition of FPS was examined by presenting the acoustic startle probe during and after each CS-US pairing trial, allowing for a trial-by-trial measurement of experience-dependent startle plasticity. In this novel protocol, mice showed robust acquisition (larger acoustic startle reflex in the presence of the CS) of FPS after as few as eight CS-US pairings. FPS was significantly greater when the CS and US were paired explicitly (light-paired) as compared to when both the US and CS were presented randomly (unpaired), or when the CS was presented alone (no shock), indicating pairing-dependent learning of the CS. Second, the present study assessed the sensitivity of FPS in mice to anxiolytic drugs. The GABA-A receptor agonists diazepam (3 and 6 mg/kg) and chlordiazepoxide (10 mg/kg) significantly reduced the expression of FPS post-training, as did the serotonin 1A receptor partial agonist buspirone (5 and 10 mg/kg). Furthermore, all three anxiolytic drugs reduced startle responding in a cue-specific manner and without significant changes in baseline responding. These data demonstrate a novel method of studying acquisition of FPS, and support the predictive validity of the FPS model of anxiolytic drug action in mice. PMID- 12655311 TI - Modulation of passive avoidance in mice by the 5-HT1A receptor agonist flesinoxan: comparison with the benzodiazepine receptor agonist diazepam. AB - The effects of the 5-HT(1A) receptor agonist flesinoxan on passive avoidance in mice were compared with those of the benzodiazepine receptor agonist diazepam. In preliminary experiments, the retention latency to enter a dark compartment in mice subjected to single-training sessions with 0.6-mA electric foot shocks for 4, 8, or 16 s slightly increased in all of the test sessions (immediately, 24 h, and 1 week after the training sessions), but none of these changes were significant. In contrast, mice subjected to double-training sessions with 0.6-mA electric foot shocks for 16 s showed a significant increase in retention latency in all of the test sessions. Pretreatment with either flesinoxan or diazepam 30 min before the double-training sessions with 0.6-mA electric foot shocks for 16 s significantly decreased the retention latency in test sessions 24 h and 1 week later. In contrast, mice pretreated with flesinoxan 24 h before the single training sessions with 0.6-mA electric foot shocks for 4, 8, or 16 s showed a significant increase in retention latency in the test sessions 24 h and/or 1 week later. Similar enhancements of retention latency in the test sessions 24 h and/or 1 week later were observed also in mice pretreated with flesinoxan 24 h before the double-training sessions. However, in this time interval following injection, pretreatment with diazepam did not affect the retention latency of mice in any of the test sessions. Neither flesinoxan nor diazepam, at the same doses and time intervals used in the passive avoidance study, modified the thresholds for flinching and jumping elicited by electrical stimuli. These results suggest that the activation of 5-HT(1A) receptors, but not benzodiazepine receptors, has a dual effect on the formation of learning and memory for an aversive event that depends on the time interval following receptor activation. PMID- 12655312 TI - Effects of strain, novelty, and NMDA blockade on auditory-evoked potentials in mice. AB - People with schizophrenia exhibit impaired ability to modify electroencephalographic event-related potential (ERP) responses to novel stimuli. These deficits serve as a window into the abnormalities of neuronal organization and function and are thought to reflect a component of genetic vulnerability for schizophrenia. We describe differences among inbred mouse strains for ERPs following a novelty detection paradigm, as a model for genetic contributions to disease vulnerability. Auditory-evoked potentials were recorded during an auditory oddball task in nonanesthetized C57BL/6J, C3H/HeJ, and DBA/2J mice prior to and following ketamine (10 mg/kg). Stimuli consisted of 80 sets of 24 standard tones followed by one novel tone. Principal component analysis yielded four temporal components that contribute to the auditory ERP responses to standard and novel stimuli. Two principal components that varied between standard and novel stimuli also differed among inbred mouse strains. Post hoc analyses indicate that strain effects on novelty detection are due to a significant difference between the response to novel and standard tones in C3H/HeJ mice that is absent in the other two strains. Inbred strains of mice vary in their ability to perform neuronal detection of change in the auditory environment. The ability to model novelty detection deficits in mice will aid in identifying genetic contributions to abnormal neuronal organization in people with schizophrenia. PMID- 12655313 TI - Effects of long-term acetyl-L-carnitine administration in rats--II: Protection against the disrupting effect of stress on the acquisition of appetitive behavior. AB - Long-term acetyl-L-carnitine (ALCAR) administration prevents the development of escape deficit produced by acute exposure to unavoidable stress. However, it does not revert the escape deficit sustained by chronic stress exposure. Rats exposed to chronic stress show a low dopamine (DA) output in the nucleus accumbens shell (NAcS) and do not acquire an appetitive behavior sustained by the earning of vanilla sugar (VS) made contingent on the choice of one of the two divergent arms of a Y-maze (VS-sustained appetitive behavior, VAB), while control rats consistently do. The present study shows that ALCAR treatment in rats exposed to a 7-day stress protocol prevented a decrease in DA output in the NAcS and medial prefrontal cortex (mPFC) of rats, and that it strengthened the DA response to VS consummation in the same two areas. Moreover, rats treated with long-term ALCAR or exposed to chronic stress while treated with ALCAR acquired VAB as efficiently as control rats. Moreover, VAB acquisition in stressed rats treated with ALCAR coincided with the reversal of the deficits in escape and in dopaminergic transmission in the NAcS. Thus, repeated ALCAR treatment preserved the DA response to VS in chronically stressed rats and this effect appeared to be predictive of the rat's competence to acquire VAB. PMID- 12655314 TI - Effect of agomelatine in the chronic mild stress model of depression in the rat. AB - Chronic mild stress (CMS), a well-validated model of depression, was used to study the effects of the melatonin agonist and selective 5-HT(2C) antagonist agomelatine (S 20098) in comparison with melatonin, imipramine, and fluoxetine. All drugs were administered either 2 h before (evening treatment) or 2 h after (morning treatment) the dark phase of the 12-h light/dark cycle. Chronic (5 weeks) evening treatment with agomelatine or melatonin (both at 10 and 50 mg/kg i.p.) dose-dependently reversed the CMS-induced reduction in sucrose consumption. The magnitude and time course of the action of both drugs was comparable to that of imipramine and fluoxetine (both at 10 mg/kg i.p.); however, melatonin was less active than agomelatine at this dose. The effect of evening administration of agomelatine and melatonin was completely inhibited by an acute injection of the MT(1)/MT(2) antagonist, S 22153 (20 mg/kg i.p.), while the antagonist had no effect in animals receiving fluoxetine or imipramine. When the drugs were administered in the morning, agomelatine caused effects similar to those observed after evening treatment (with onset of action faster than imipramine) but melatonin was ineffective. Moreover, melatonin antagonist, S 22153, did not modify the intakes in stressed animals receiving morning administration of agomelatine and in any other control and stressed groups tested in this study. These data demonstrate antidepressant-like activity of agomelatine in the rat CMS model of depression, which was independent of the time of drug administration. The efficacy of agomelatine is comparable to that of imipramine and fluoxetine, but greater than that of melatonin, which had no antidepressant-like activity after morning administration. While the evening efficacy of agomelatine can be related to its melatonin receptors agonistic properties, its morning activity, which was not inhibited by a melatonin antagonist, indicates that these receptors are certainly required, but not sufficient to sustain the agomelatine efficacy. It is therefore suggested that the antidepressant-like activity of agomelatine depends on some combination of its melatonin agonist and 5-HT(2C) antagonist properties. PMID- 12655315 TI - Association between estrus cycle-related changes in respiration and estrus cycle related aggression in outbred female Wistar rats. AB - Premenstrual dysphoric disorder is characterized by irritability surfacing during the luteal phase of the menstrual cycle, and disappearing shortly after the onset of menstruation. Although the cardinal symptoms of premenstrual dysphoria are different from those of panic disorder, the two conditions share a number of traits indicating that they both may be associated with abnormalities in the regulation of respiration. Both subjects with panic disorder and subjects with premenstrual dysphoria are hence reported to display enhanced respiratory variability, and to experience anxiety when exposed to CO(2). In the present study, the possible influence of the estrus cycle on respiratory parameters in outbred female rats of the Wistar strain was investigated. Before being tested with respect to respiration, the rats were subdivided into two groups: those displaying estrus cycle-related variation in aggression, as evaluated using the resident intruder paradigm, and those not showing aggression throughout the cycle. Whereas the former group was found to display higher respiratory rate during the diestrus phase than during the proestrus/estrus phase, no cycle related variation in respiration was observed in animals not showing cycle related variation in aggression. The results support previous studies indicating that the estrus cycle exerts an influence on respiration, and suggest that rats prone to cycle-related aggression are more sensitive also to the influence of hormonal cyclicity on respiration. The possible bearing of these findings for the aberration in respiration displayed by subjects with premenstrual dysphoria is discussed. PMID- 12655316 TI - Pronounced cognitive deficits following an intravenous L-tryptophan challenge in first-degree relatives of bipolar patients compared to healthy controls. AB - Cognitive impairment has repeatedly been described in bipolar disorders (BD). Serotonin (5-hydroxytryptophan; 5-HT) is possibly involved in these cognitive processes, more particularly in executive functions, learning, memory, and attention. The aim of this study was to investigate serotonergic vulnerability and its relation to cognitive functioning in healthy first-degree relatives of BD patients. We investigated the effects of an intravenous (i.v.) tryptophan (Trp) challenge and placebo on cognitive performance in 30 healthy first-degree relatives of bipolar patients (FH) and 15 matched controls in a double-blind crossover design. A distinction was made between relatives of type I BD patients (FH I) and type II BD patients (FH II). Performances on planning, memory, attention, and psychomotor tasks were assessed 3 h after Trp infusion. After Trp, planning and attention were impaired in FH subjects but not in controls. Independent of Trp, FH subjects showed cognitive deficits on memory, focused and divided attention, and psychomotor performance. FH I subjects showed more pronounced cognitive impairments then FH II and controls. In all groups, Trp impaired memory and psychomotor performance significantly. In conclusions, cognitive deficits in FH following Trp may reflect a central 5-HT vulnerability in frontal brain areas. Independent of Trp, cognitive deficits in FH provide evidence for a trait marker for BD. PMID- 12655317 TI - Neurotrophic effects of electroconvulsive therapy: a proton magnetic resonance study of the left amygdalar region in patients with treatment-resistant depression. AB - Negatively balanced neurotrophic factors may be important in precipitating clinical depression. Recently, it has been reported that antidepressant therapy may exert positive neurotrophic effects. The aim of this study was to detect probable neurotrophic changes during electroconvulsive therapy (ECT). For this purpose, N-acetylaspartate (NAA), an amino acid exclusively located in neurons, and other brain metabolites such as glutamine/glutamate (Glx), choline (Cho), and creatine (Cr) were measured in patients by localized proton magnetic resonance spectroscopy. A total of 28 severely depressed patients (DSM-IV) were enrolled, and the left amygdalar region was investigated by proton STEAM spectroscopy before and after unilateral ECT. The results were compared with 28 age- and gender-matched controls using nonparametric paired and unpaired tests. A significant increase in NAA was observed only in ECT responders (n=14; p=0.019). Five out of 14 nonresponders to ECT monotherapy were remeasured following a clinical improvement after continued ECT combined with antidepressants and were then found also to present a significant increase in NAA. In all successfully treated patients, parallel observations, that is, increased levels, were made for Glx, whereas Cho and Cr were unchanged. In conclusion, our preliminary finding of increased NAA concentrations after successful ECT may indicate a probable neurotrophic effect of ECT. PMID- 12655318 TI - Extended amygdala and emotional salience: a PET activation study of positive and negative affect. AB - Functional neuroimaging studies have implicated amygdaloid and basal forebrain regions, including sublenticular extended amygdala (SLEA), in the mediation of aversive emotional responses. However, it is not clear whether SLEA responds to 'aversiveness' or to general stimulus salience. We predicted that both pleasant and aversive stimuli would activate this region. Using [(15)O] water PET, we studied 10 healthy subjects while viewing pleasant, aversive, neutral, and blank images. Each subject underwent eight scans, which were processed and averaged with standard statistical methods. Both positive and negative stimuli activated regions in SLEA. Both positive and negative content activated the visual cortex, relative to neutral content. Aversive stimuli deactivated the left frontal pole, relative to positive and neutral stimuli. These findings demonstrate that both positive and negative emotional content evokes processing in the sublenticular/extended amygdala region, suggesting that this region is involved in general emotional processing, such as detection or attribution of salience. PMID- 12655319 TI - Circadian clock-related polymorphisms in seasonal affective disorder and their relevance to diurnal preference. AB - Disturbed circadian rhythms have been observed in seasonal affective disorder (SAD). The aim of this study was to further investigate this connection, and to test for potential association between polymorphisms in circadian clock-related genes and SAD, seasonality (seasonal variations in mood and behavior), or diurnal preference (morningness-eveningness tendencies). A total of 159 European SAD patients and 159 matched controls were included in the genetic analysis, and subsets were screened for seasonality (n=177) and diurnal preference (n=92). We found that diurnal preference was associated with both SAD and seasonality, supporting the hypothesis of a link between circadian rhythms and seasonal depression. The complete case-control material was genotyped for polymorphisms in the CLOCK, Period2, Period3, and NPAS2 genes. A significant difference between patients and controls was found for NPAS2 471 Leu/Ser (chi(2)=9.90, Bonferroni corrected P=0.035), indicating a recessive effect of the leucine allele on disease susceptibility (chi(2)=6.61, Bonferroni corrected P=0.050). Period3 647 Val/Gly was associated with self-reported morningness-eveningness scores (n=92, one-way ANOVA: F=4.99, Bonferroni corrected P=0.044), with higher scores found in individuals with at least one glycine allele (t=3.1, Bonferroni corrected P=0.013). A second, population-based sample of individuals selected for high (n=127) or low (n=98) degrees of seasonality, was also genotyped for NPAS2 471 Leu/Ser. There was no significant difference between these seasonality extreme groups, and none of the polymorphisms studied were associated with seasonality in the SAD case-control material (n=177). In conclusion, our results suggest involvement of circadian clock-related polymorphisms both in susceptibility to SAD and diurnal preference. PMID- 12655320 TI - Alcohol hangover effects on memory functioning and vigilance performance after an evening of binge drinking. AB - The impairing effects on memory functioning after acute alcohol intoxication in healthy volunteers and after chronic use in alcoholics are well established. However, research determining the next-morning effects of a single episode of binge drinking on memory functioning is scarce. A total of 48 healthy volunteers participated in a single-blind study comprising an evening (baseline) session, followed by a treatment administration (ethanol 1.4 g/kg or placebo), and a morning session. Memory was tested with a word-learning test (including immediate and delayed recall, and recognition). Further, a 45-min Mackworth clock test for measuring vigilance was included (parameters: number of hits and false alarms) and subjective alertness was assessed, to infer whether word-learning test findings reflect sedation or specific memory impairments. Delayed recall in the morning session was significantly worse in the alcohol group when compared to the placebo group (F(1,42)=6.0, p<0.02). In contrast, immediate recall and recognition were unimpaired in the alcohol group. In the morning session, relative to the placebo group, subjective alertness was significantly reduced in the alcohol group before and after the tests (F(1,44)=8.7, p<0.005; F(1,44)=13.3, p&<0.001, respectively). However, in the Mackworth clock test, the alcohol group and placebo group did not differ significantly in the morning session. The specific findings of impaired delayed recall show that memory retrieval processes are significantly impaired during alcohol hangover. Vigilance performance was not significantly affected, indicating that this memory impairment does not reflect sedation. PMID- 12655321 TI - Alcohol reduces prefrontal cortical excitability in humans: a combined TMS and EEG study. AB - The effects of alcohol (0.8 g/kg) on the prefrontal cortex were studied in nine healthy subjects using the technique of transcranial magnetic stimulation (TMS) combined with electroencephalography (EEG). A total of 120 magnetic pulses were delivered with a figure-of-eight coil to the left prefrontal cortex at the rate of 0.4-0.7 Hz. The EEG was recorded simultaneously with 60 scalp electrodes (41 electrodes were used for analysis); the TMS-evoked activation was estimated by the area under the global mean field amplitude (GMFA) time curve. TMS caused changes in EEG activity lasting up to 270 ms poststimulus. Alcohol decreased GMFA at 30-270 ms poststimulus (713+/-303 vs 478+/-142 microV ms; p=0.007). Alcohol induced differences were most pronounced at anterior electrodes. These results suggest that alcohol reduces the excitability in the prefrontal cortex. PMID- 12655322 TI - A clinical laboratory paradigm for evaluating medication effects on alcohol consumption: naltrexone and nalmefene. AB - Opiate antagonist medications have been shown to improve alcoholism treatment, but few human laboratory-based studies investigating mechanisms for these effects have been conducted on alcohol dependent persons. The present study was designed to determine the impact of two opiate antagonists on alcohol consumption among nontreatment-seeking alcoholics (n=125) and social drinkers (n=90). Participants were randomly assigned to receive placebo, naltrexone (titrated to 50 mg/day), or nalmefene (titrated to 40 mg/day) for 8 days with an alcohol laboratory session on the final day. Alcohol consumption was monitored in the natural environment during the first 5 medication days, and during a choice consumption paradigm following a standard 'priming' alcohol dose in a bar-laboratory setting. Social drinkers consumed less alcohol than alcoholics during the prelab medication period and the laboratory choice consumption paradigm, and they attained lower blood alcohol levels than alcoholics following the priming drink. Both opiate antagonist medications equally reduced drinking amounts and frequency among alcoholics but not social drinkers, relative to placebo, during natural environment and bar-lab alcohol consumption evaluations. Greater medication side effects, mostly mild in nature, were observed in participants taking nalmefene. These findings demonstrate that both naltrexone and nalmefene can lead to reductions in alcohol consumption among alcoholics who are not attempting to reduce drinking. Similar laboratory paradigms may offer substantial advantages for observing these effects during evaluation of other medications as well. PMID- 12655323 TI - Intravenous nicotine reduces cerebral glucose metabolism: a preliminary study. AB - Nicotine is self-administered by smoking tobacco products, and enhances positive mood (at least in smokers). Since most drugs of abuse decrease regional cerebral metabolic rate(s) for glucose (rCMRglc) in human subjects, we posited that administration of nicotine would similarly reduce rCMRglc. Positron emission tomography (PET) with [F-18]fluorodeoxyglucose was used to assess the effects of intravenous nicotine (1.5 mg) on cerebral glucose metabolism in six healthy male volunteers (21-38 years of age). Two PET assays (placebo and nicotine) were performed, and subjective self-reports of mood and feeling state were collected. Data were analyzed using analysis of variance. Nicotine reduced global glucose metabolism (by 9.51% of placebo control), with reductions in most of the 30 individual regions tested. Nine regions had bilateral effects that reached statistical significance (p&<0.05, uncorrected for the number of regions tested), although the statistical model used did not separate these effects from a global effect. The subjects reported both positive and negative effects of nicotine on mood/feeling state. The widespread decreases in cerebral metabolism are consistent with the many effects of nicotine on cognition and mood. The findings indicate that nicotine resembles other drugs of abuse in reducing brain metabolism, perhaps by a common mechanism. PMID- 12655324 TI - Development of an abbreviated schizophrenia quality of life scale using a new method. AB - The goal of the study was to develop and apply a predictive model approach to reduce the number of items collected for scales that yield a total summary score. A parsimonious subset of items from the 21-item Quality of Life Scale (QLS) that can accurately predict the total scale score was sought and evaluated in 198 patients with schizophrenia, using a statistical modeling approach. Two additional data sets were used for model validation: the subset of 101 patients used in the model construction who had the QLS administered approximately 1 year later and a new sample of 37 patients. Using only seven QLS items as predictors, the correlation was 0.9831 between the predicted and true QLS totals. Applying the model based for these seven QLS items, the correlations from the first and second validation data sets were 0.9791 and 0.9637, respectively. The study demonstrates that a small subset of items of the QLS predicts the entire 21-item scale with high accuracy. Two validation samples have confirmed the finding. This reduces the effort associated with scale administration and is likely to increase the assessment of an important functional domain. Such models can guide efforts for item reduction in other rating instruments. PMID- 12655325 TI - Influence of benzodiazepines on auditory perception. AB - The aim of this study was to test for an influence of benzodiazepine (BZD) on various perceptual and/or cognitive auditory processes. Loudness, auditory selective attention, and the ability of subjects to form perceptual streams out of alternating tone sequences were tested. Nine subjects were tested before, 1, 3, 7, and 24 h after a single-dose oxazepam vs placebo administration in a crossover design. A sample of blood allows us to measure plasma oxazepam concentration. The results revealed a significant reduction in stream segregation expressed as d' scores 1 h after oxazepam intake in the test subjects. No significant change occurred across time in the same subjects when they were administrated a placebo in another session. Furthermore, oxazepam had no substantial and systematic influence either on auditory selective attention or on loudness perception. Altogether, these results suggest that the perceptual organization of sound sequences involves inhibitory neural mechanisms, which can be affected by BZDs. This outcome is consistent with existing models of auditory stream segregation and may be paralleled with earlier findings on the effect of BZDs on perceptual binding in the visual modality. PMID- 12655326 TI - Subchronic haloperidol downregulates dopamine synthesis capacity in the brain of schizophrenic patients in vivo. AB - The antipsychotic effect of neuroleptics cannot be attributed entirely to acute blockade of postsynaptic D(2)-like dopamine (DA) receptors, but may arise in conjunction with the delayed depolarization block of the presynaptic neurons and reduced DA synthesis capacity. Whereas the phenomenon of depolarization block is well established in animals, it is unknown if a similar phenomenon occurs in humans treated with neuroleptics. We hypothesized that haloperidol treatment should result in decreased DA synthesis capacity. We used 6-[(18)F]fluoro-L-dopa (FDOPA) and positron emission tomography (PET) in conjunction with compartmental modeling to measure the relative activity of DOPA decarboxylase (DDC) (k(D)(3), min(-1)) in the brain of nine unmedicated patients with schizophrenia, first in the untreated condition and again after treatment with haloperidol. Patients were administered psychometric rating scales at baseline and after treatment. Consistent with our hypothesis, there was a 25% decrease in the magnitude of k(D)(3) in both caudate and putamen following 5 weeks of haloperidol therapy. In addition, the magnitudes of k(D)(3) in cerebral cortex and thalamus were also decreased. Psychopathology as measured with standard rating scales improved significantly in all patients. The decrease of k(D)(3) in the thalamus was highly significantly correlated with the improvement of negative symptoms. Subchronic treatment with haloperidol decreased the activity of DDC in the brain of patients with schizophrenia. This observation is consistent with the hypothesis that the antipsychotic effect of chronic neuroleptic treatment is associated with a decrease in DA synthesis, reflecting a depolarization block of presynaptic DA neurons. We link an alteration in cerebral catecholamine metabolism in human brain with the therapeutic action of neuroleptic medication. PMID- 12655327 TI - Schizophrenia subjects show intact success-related neural activation but impaired uncertainty processing during decision-making. AB - Decision-making is a complex process that is important for everyday life. This study examined the effect of the degree of success, and outcome uncertainty, on decision-making and associated neural substrate activation in schizophrenia subjects (SZS) and normal comparison subjects (NCS). A total of 15 subjects with a diagnosis of schizophrenia and 15 age- and education-matched NCS participated in this study. These subjects completed the two-choice prediction task during functional magnetic resonance imaging. Decision-making characteristics and activation of neural substrates were obtained at 20, 50, or 80% error rate. Success and uncertainty influenced the behavioral characteristics on the two choice prediction task, and the task-related activation in SZS and NCS. Neither success nor uncertainty differentially affected the behavioral characteristics of SZS relative to NCS during the two-choice prediction task. Nonetheless, there was a significant interaction between group and error rate in bilateral parietal cortex. The activation in NCS was the highest when the outcome was most uncertain. In contrast, task-related activation in SZS was not modulated by the degree of uncertainty. Thus, SZS failed to utilize the parietal cortex to process decision-making situations with highly uncertain outcomes. PMID- 12655328 TI - Differential effects of antidepressants on glucocorticoid receptors in human primary blood cells and human monocytic U-937 cells. AB - A number of data support the assumption that antidepressants (ADs) normalize the altered function of the hypothalamic-pituitary-adrenocortical (HPA) system involved in the pathophysiology of depressive disorder via direct effects on glucocorticoid receptors (GRs). In the present study, we examined the tricyclic ADs desipramine (DESI) and imipramine (IMI), the noradrenaline reuptake inhibitor maprotiline (MAPRO), and the noradrenergic and specific serotonergic AD (NaSSA) mirtazapine (MIR) for their effects on GR expression in primary human leukocytes and in monocytic U-937 cells. Semiquantitative RT-PCR indicated that the ADs exert differential effects on GR-mRNA levels in both primary human leukocytes and U-937 cells: whereas MAPRO and IMI did not induce pronounced changes in GR-mRNA levels, DESI and MIR significantly decreased the amounts of GR-mRNA in both cell systems. Further characterization of the effects of MIR revealed a time dependency of the regulation with an initial increase of GR-mRNA levels above control levels after 2.5 h of treatment and a decrease after 4, 24, and 48 h of incubation. A dose-response analysis demonstrated maximal effects of MIR at a concentration of 10(-7) M. Immunohistochemical studies showed that MIR increased the GR protein levels in a time-dependent manner and that this upregulation appeared earlier by additional treatment with dexamethasone (DEX). A translocation of the GR protein from the cytoplasm to the nucleus was induced between 24 and 48 h of treatment with MIR and MIR/DEX, respectively. Taken together, our data further support the assumption that ADs influence the neuroendocrine and immune system via effects on cellular GRs. PMID- 12655329 TI - Inappropriate choice of the experimental unit leads to a dramatic overestimation of the significance of quantitative trait loci for prepulse inhibition and startle response in recombinant congenic mice. PMID- 12655335 TI - Hepatitis C: virus, host, disease. PMID- 12655336 TI - Subversion of cell survival and cell death: viruses as enemies, tools, teachers and allies. PMID- 12655337 TI - Hepatitis C viral kinetic models. PMID- 12655338 TI - Intrahepatic natural immunity and HCV immunopathogenesis. PMID- 12655339 TI - Does nitric oxide play a pathogenic role in hepatitis C virus infection? PMID- 12655340 TI - Transgenic models for Hepatitis C virus pathogenesis. PMID- 12655341 TI - Mitogenesis by ligands of nuclear receptors: an attractive model for the study of the molecular mechanisms implicated in liver growth. PMID- 12655342 TI - HCV and interferon: viral strategies for evading innate defence mechanisms in the virus-host battle. PMID- 12655343 TI - HIV/HCV co-infection: putting the pieces of the puzzle together. PMID- 12655344 TI - Hepatitis C virus biology. AB - Hepatitis C virus infection represents a major problem of public health with around 350 millions of chronically infected individuals worldwide. The frequent evolution towards severe liver disease and cancer are the main features of HCV chronic infection. Antiviral therapies, mainly based on the combination of IFN and ribavirin can only assure a long term eradication of the virus in less than half of treated patients. The mechanisms underlying HCV pathogenesis and persistence in the host are still largely unknown and the efforts made by researchers in the understanding the viral biology have been hampered by the absence of a reliable in vitro and in vivo system reproducing HCV infection. The present review will mainly focus on viral pathogenetic mechanisms based on the interaction of HCV proteins (especially core, NS3 and NS5A) with host cellular signaling transduction pathways regulating cell growth and viability and on the strategies developed by the virus to persist in the host and escape to antiviral therapy. Past and recent data obtained in this field with different experimental approaches will be discussed. PMID- 12655345 TI - HCV immunology--death and the maiden T cell. AB - Cellular immune responses play an important role in the control of hepatitis C virus (HCV), although in the majority of cases they ultimately fail. We examine the mechanisms by which virus-specific T cells may interact with a cell that is infected with HCV and how this interaction may explain the success and failure of the immune response. As an infected cell presenting foreign antigen, the hepatocyte will interact with a large number of lymphocytes, both by direct cell to cell contact and by indirect means through the secretion of cytokines and chemokines. These interactions may lead on the one hand to the death of infected hepatocytes or suppression of viral replication and on the other hand to the death of T lymphocytes or down regulation of their function. We suggest that activation of lymphocytes in lymphoid organs leads to generation of effector T cells (positive loop), while at the same time presentation of antigen in the liver either on hepatocytes or other specialised antigen presenting cells depresses these responses (negative loop). This model helps to explain both the specific phenotype and low frequencies of HCV specific CTL in chronic infection, through early elimination of cells before expansion and maturation can occur. The outcome of HCV infection is likely to result from the early balance between these two simultaneous loops. PMID- 12655346 TI - Apoptosis in hepatitis C virus infection. AB - Infection with hepatitis C virus (HCV) is characterized by inflammatory liver damage and a long viral persistence associated with an increased risk of developing hepatocellular carcinoma. Both in liver damage and in oncogenesis a disturbance of apoptosis has been implicated, although the underlying mechanisms in these apparently opposite processes are incompletely understood. HCV-triggered liver injury is mediated mainly by host immune mechanisms and eventually by direct cytopathic effects of HCV. Recent data shows that caspase activation, either triggered by death ligands, other cytokines, granzyme B or HCV proteins, is considerably upregulated in HCV-infected liver. Interestingly, caspase activation appears to correlate closely with the inflammatory response. Data about the role of single HCV proteins, either in cultured cells or transgenic animals models, however, are contradictory, as both pro- and anti-apoptotic effects have been observed. Nevertheless, apoptosis induction upon HCV infection may critically contribute to liver damage, while inhibition of apoptosis may result in HCV persistence and development of hepatocellular carcinoma. PMID- 12655347 TI - Hepatitis C and liver fibrosis. AB - Chronic hepatitis C progresses to cirrhosis within 20 years in an estimated 20 30% of patients, while running a relatively uneventful course in most others. Certain HCV proteins, such as core and NS5A, can induce derangement of lipid metabolism or alter signal transduction of infected hepatocytes which leads to the production of reactive oxygen radicals and profibrogenic mediators, in particular TGF-beta1. TGF-beta1 is the strongest known inducer of fibrogenesis in the effector cells of hepatic fibrosis, i.e. activated hepatic stellate cells and myofibroblasts. However, fibrogenesis proceeds only when additional profibrogenic stimuli are present, e.g. alcohol exposure, metabolic disorders such as non alcoholic steatohepatitis, or coinfections with HIV or Schistosoma mansoni that skew the immune response towards a Th2 T cell reaction. Furthermore, profibrogenic polymorphisms in genes that are relevant during fibrogenesis have been disclosed. This knowledge will make it possible to identify those patients who are most likely to progress and who need antiviral or antifibrotic therapies most urgently. However, even the best available treatment, the combination of pegylated interferon and ribavirin, which is costly and fraught with side effects, eradicates HCV in only 50% of patients. While the suggestive antifibrotic effect of interferons (IF-gamma>alpha,beta), irrespective of viral elimination, has to be proven in randomised prospective studies, additional, well tolerated and cost-effective antifibrotic therapies have to be developed. The combination of cytokine strategies, e.g. inhibition of the key profibrogenic mediator TGF-beta, with other potential antifibrotic agents appears promising. Such adjunctive agents could be silymarin, sho-saiko-to, halofuginone, phosphodiesterase inhibitors, and endothelin-A-receptor or angiotensin antagonists. Furthermore, drug targeting to the fibrogenic effector cells appears feasible. Together with the evolving validation of serological markers of hepatic fibrogenesis and fibrolysis an effective and individualised treatment of liver fibrosis is anticipated. PMID- 12655348 TI - Viral versus cellular BCL-2 proteins. AB - All gamma herpesviruses and a few other viruses encode at least one homologue of the mammalian cell death inhibitor BCL-2. Gamma herpesviruses are associated with human and animal lymphoid and epithelial tumours. However, the role of these viral BCL-2 homologues in the virus replication cycle or in human disease is not known, though recent developments show progress in this area. The structure of viral BCL-2 family protein, KSBcl-2, is similar to that of cellular family members, but viral BCL-2 proteins differ functionally from the cellular proteins, apparently escaping the regulatory mechanisms to which their cellular counterparts are subjected. Thus, exploring the biochemical and biological functions of the viral BCL-2 family proteins will increase our understanding of their role in virus infections and will undoubtedly teach us something about their cellular kin. PMID- 12655349 TI - Trying to catch the HCV virus in its 'battle field'. PMID- 12655350 TI - Tissue transglutaminase in HCV infection. PMID- 12655351 TI - Capacitance and resistance of the bilayer lipid membrane formed of phosphatidylcholine and cholesterol. AB - Capacity and electric resistance of lipid membranes composed of lecithin and cholesterol were determined. The components were chosen for the study because they were present in biological membranes. Capacitance of the lecithin and cholesterol membranes amounts to 0.38 and 0.61 microF/cm(2), and resistance to 1.44(10(4)and 2.12(10(6)Omega cm(2), respectively. A 1:1 complex appears as a result of lecithin-cholesterol membrane formation. Parameters of the membrane formed of the lecithin-cholesterol complex were determined: surface concentration (Gamma(3)), capacitance (C(3)), and conductance (R;(3)(-1), as well as the stability constant (K) of the complex. The mean values of those magnitudes are as follows: 4.265(10(-6)mol/m(2), 0.54 microF/cm(2), 1.381(10(-6)Omega(-1)cm(-2)and 3.748(10(7), respectively. PMID- 12655352 TI - Lysosomal high molecular weight multienzyme complex. AB - Three acidic glycosidases: beta-galactosidase (beta-GAL, EC 3.2.1.23), alpha neuraminidase (NEUR, sialidase, EC 3.2.1.18), N-acetylaminogalacto-6-sulfate sulfatase (GALNS, EC 3.1.6.4) and serine carboxypepidase cathepsin A (EC 3.4.16.1) form a functional high molecular weight complex in the lysosomes. The major constituent of this complex is cathepsin A, the so-called "lysosomal protective protein" (PPCA). By forming a multienzyme complex, it protects the glycosidases from rapid intralysosomal proteolysis, and it is also required for the intracellular sorting and proteolytic processing of their precursors. In man, a deficiency of cathepsin A leads to a combined deficiency of beta-GAL and NEUR activities, called "galactosialidosis". Multiple mutations identified in the cathepsin A gene are the molecular basis of this lysosomal storage disease. This review describes the structural organization of the lysosomal high molecular weight multienzyme complex and the importance of the protective protein/cathepsin A in physiology and pathology. PMID- 12655354 TI - Disturbances of stem circumnutations evoked by wound-induced variation potentials in Helianthus annuus L. AB - The relationship between evoked electrical activity and stem movements in three week old sunflowers was demonstrated. Electrical potential changes (recorded by Ag/AgCl extracellular electrodes) and time-lapse images (from a top view camera) were recorded and analyzed. A heat stimulus applied to the tip of one of the second pair of leaves evoked a variation potential, transmitted basipetally along one side of the stem. After stimulation, disturbances of circumnutations occurred. They included: changes in the period, disorders in the elliptical shape, and, in some cases, reversion of direction (of movement). We suggest that asymmetrically propagated variation potential induces asymmetric stem shrinking and bending, which strongly disturbs circumnutations. Our results confirm the involvement of electrical potential changes in the mechanism of stem nutations. PMID- 12655353 TI - Damage to the erythrocyte membrane caused by chlorophenoxyacetic herbicides. AB - We studied the damage caused to erythrocyte membranes by chlorophenoxyacetic herbicides. An increase in haemolysis was observed. The compounds investigated caused lipid bilayer damage by lipid peroxidation, as well as an increase in membrane fluidity at the 16th carbon atom of fatty acids was observed. Metabolites caused damage to membrane proteins - the free SH group content was increased. Higher toxicity of metabolites compared to basic compounds was observed. PMID- 12655355 TI - Characterization of the human lactoferrin (HLF) cell line HLFK1, generated in CBA mice. AB - A human lactoferrin-specific cell line was generated in CBA mice, sensitized with 200 microg HLF in Freund's complete adjuvant. HLFK1 cells derived from the lymph nodes of these mice were maintained using HLF as the antigen. HLF was added at the beginning of each 14-day restimulation cycle, at a concentration of 100 microg/ml. The presentation of the antigen to HLFK1 was demonstrated using glass adherent lymphocytes from spleens (GAL) as the antigen-presenting cells (APC). The presentation of HLF by GAL was highly efficient; a very low concentration of the antigen (1 microg/ml) was enough to stimulate proliferation of the HLFK1 cell line. HLFK1 did not proliferate in the presence of ovalbumin or bovine lactoferrin (BLF), which is structurally related to HLF. However, we found that BLF caused a reduction in the proliferation of the HLFK1 cell line when BLF was added to the cultures together with the antigen - (HLF). On the other hand, proliferation of the HLFK1 cell line was not inhibited by pretreatment of the antigen-presenting cells or T cells with BLF. Therefore, we suggest that bovine lactoferrin may interfere with the binding or uptake of the antigen (HLF). Alternatively, BLF may nonspecifically inhibit the activation of the HLFK1 cell line. PMID- 12655356 TI - Peroxisome proliferator-activated receptor alpha is downregulated in the failing human heart. AB - Cardiac hypertrophy in humans is associated with a decrease in myocardial fatty acid beta-oxidation (FAO) and accompanying alterations in metabolic gene expression. Flux through the cardiac FAO pathway, which is the principal source of energy production in the adult mammalian heart, is tightly controlled in accordance with energy demands. In rodents, the FAO pathway is under control of a nuclear peroxisome proliferator-activated receptor alpha (PPARalpha?. We sought to delineate the molecular regulatory events involved in the energy substrate preference switch from fatty acids to glucose during cardiac hypertrophic growth in humans. We analysed the amount of PPARalpha protein in human cardiac tissue. PPARalpha protein level was measured in homogenates prepared from left ventricular biopsies taken from five control donor hearts and compared to the amount of this transcription factor in biopsies from five patients with compensated end-stage heart failure (HF) at the time of transplantation. Using Western blot analysis with a monoclonal antibody against human PPARalpha, we observed a significant decrease (54%) in the mean amount of PPARalpha in the group of HF patients compared to that in the donor tissue. This study indicates that the decrease in cardiac PPARalpha transcription factor gene expression observed in the failing human heart could play an important role in a reduction in fatty acid utilisation by the adult heart during cardiac hypertrophy. PMID- 12655357 TI - The hemolytic and physiological activities of mixtures of some phenoxy and organophosphorous herbicides. AB - Experiments were performed investigating the potential to improve the biological activity of some phenoxy and organophosphorous compounds by using them in binary mixtures. The compounds were: 2,4-dichlorophenoxyacetic acid (1) and its sodium salt (2), dibutyl 1-butylamino-1-cyclohexanephosphonate (3) and diethyl 9 butylamino-9-fluorenephosphonate (4), all widely used as herbicides. There were two test methods: the inhibition of cucumber (Cucumis sativus) growth induced by one single herbicide or by equimolar binary mixtures of herbicides; and, in parallel, the hemolytic efficiency of separate compounds or their mixtures. The hemolytic properties of the compounds were studied as hemolysis is generally a good measure of their toxicity, especially in the case of lipophilic compounds. Pig erythrocytes were used as good models for the determination of toxicity and the kinetics of red blood cell hemolysis. In the plant-based experiments, binary mixtures were found to display additive type toxicity. The compounds' hemolytic activities were of additive or antagonistic types. In some combinations, the addition of a second component did not change the hemolytic efficiency of the first component, and vice versa. PMID- 12655358 TI - A morphological and immunohistochemical investigation of guinea pig skin after the introduction of substance P and VIP. AB - The aim of this study was to examine a morphological picture of guinea pig skin that had been injected with neuropeptides (NPS)(2)- substance P (SP) and guinea pig vasoactive intestinal peptide (VIP) - to elucidate their local influence. Routine histological stainings were performed, together with immunohistochemical reactions for T cells and for macrophages. In the deeper layers of the skin, T cell and macrophagic infiltrations were observed. The intensity of these changes was greater 24 hours after injections than that observed at the third hour of the experiment. PMID- 12655359 TI - The effect of a selective inhibition of potassium channels on the relaxation induced by nitric oxide in the human pregnant myometrium. AB - The aim of this study was to investigate whether apamin-sensitive K(+)channels play a role in the NO induced relaxation of the human pregnant myometrium. Concentration-response curves for sodium nitroprusside (SNP) (10(-9)-10(-4)M) were constructed in the absence and presence of 10(-8)M apamin and 10(-7)M charybdotoxin (CTX). Preincubation with apamin resulted in a significant attenuation of the relaxation caused by SNP, while pre-treatment with CTX insignificantly decreased the SNP induced relaxation. Our findings suggest that apamin-sensitive K(+)channels exist in the human pregnant myometrium and play a role in modulation of the myometrium response to NO donors PMID- 12655360 TI - Associating oligonucleotides with positively charged liposomes. AB - Oligonucleotides (ODNs) are short (up to 30 bases) fragments of single-stranded nucleic acids that are used as sequence specific regulators of gene expression and anti-sense based therapeutics. ODNs are frequently aggregated with particulates in order to improve their pharmacological characteristics. Complexes of ODN and lipid aggregates are among the most commonly mentioned in the literature. In order to control the formation and final properties of such aggregates, a detailed description of how ODN interacts with the lipid surface is needed. In this paper, we present the results of fluorescence measurements regarded an association of 20 base ODN, labelled with fluorescein, and a lipid surface containing various amount of positive charge. Unilamellar lipid vesicles were formed from egg phosphatidylcholine (PC) and various amounts of the cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP). It was found that about 20 mol% of DOTAP in the lipid bilayer suffices to obtain complete ODN association. This result was further confirmed via measurements performed by fluorescence correlation spectroscopy (FCS). These in turn showed that the diffusion time of labelled ODN in the presence of cationic liposomes decreases. Also, the particle number and count rate were reduced, concurring with conclusions derived from steady-state fluorescence spectroscopy results. PMID- 12655361 TI - The nonallosteric mechanism of enzyme activity regulation. Is it the only true mechanism? AB - The nonallosteric regulation mechanism of enzyme reaction velocity assumes that the substrate and enzyme interact via a metal cation and form simple and mixed, mono- and multi-nuclear complexes. A solution of equations for individual cases gives a function of initial reaction velocity at any given substrate or modifier concentration. This function can describe kinetic effects that are considered allosteric, as well as phenomena omitted by commonly-accepted models. PMID- 12655362 TI - The effect of selenium on the accumulation of some metals in Zea mays L. plants treated with indole-3-acetic acid. AB - In this study, we examined the relationship between the accumulation of NaHSeO3, the plant hormone (IAA), and some nutrient elements (K(+), Na(+), Ca(2+)) in the tissues of the roots, mesocotyls and leaves of Zea mays L. plants. Our experiments were carried out with eight- to nine-day old maize plants (Zea mays L. var K33xF2) grown on Hoagland's medium containing the standard macro- and microelements, IAA and NaHSeO(3). The accumulation of selenium, potassium, sodium and calcium in the seedlings was measured by emission spectroscopy using a spectrometer with excitation by the argon inductively coupled plasma technique (ICP-AES). We observed that when selenite and phytohormone (IAA) are present in the external medium of growing plants, they change the uptake and accumulation of some cations (K(+), Na(+), Ca(2+)) in the leaf, mesocotyl and root tissues. The change of transport of some nutrient elements is probably one of the first observed symptoms of selenium's effects on plants. PMID- 12655363 TI - pH-dependent influence of a quaternary ammonium salt and an aminoester on the yeast Saccharomyces cerevisiae ultrastructure. AB - Quaternary ammonium salts inhibited the growth of yeast especially at pH higher (pH 8) than optimal. It was postulated that compounds integrate with the cell membrane and interfere with its functions. The yeast cell ultrastructure investigated under an electron microscope confirms this hypothesis. A relatively high percentage of cells treated at pH 6 with the quaternary ammonium salt of alanine derivative (DMALM-12) at the minimal inhibitory concentration showed an irregularity in the cell shape. No such irregularity was observed in the control. Besides, in the cells treated with the drug, practically no lipid droplets were seen at all. Inside the control cells, electron-dense round bodies were clearly seen and interpreted as vacuoles. These bodies were absent in the cells treated with DMALM-12. Although the yeast cells growing at pH 8 showed a more or less normal shape, they seemed to have difficulty in budding - no fully developed buds were found in the preparations. Only some convexities of the cell wall were seen that could be the beginning of budding which stopped early after the start. Some changes in the round bodies interpreted as vacuoles were visible: they were less dense and full of granules. PMID- 12655364 TI - The dual mechanism of the antifungal effect of new lysosomotropic agents on the Saccharomyces cerevisiae RXII strain. AB - Quinacrine was used to visualize the intracellular pH changes in the yeast strain Saccharomyces cerevisiae RXII occurring after exposure to four recently synthesized lysosomotropic drugs: DM-11, PY-11, PYG-12s and DMAL-12s. The cells took up quinacrine, mostly accumulating it in their vacuoles. DM-11 and PY-11 gave rise to diffuse quinacrine fluorescence throughout the cells, with the vacuoles staining to a somewhat greater extent than the cytosol. This quinacrine detected overall acidification of the cell interior is very probably caused by blocking of plasma membrane H(+)-ATPase. PYG-12s gave rise to a strong vacuolar accumulation of the dye. Like the vacuolar ATPase inhibitor bafilomycin A(1), DMAL-12s strongly lowered the intensity of quinacrine fluorescence. Owing to its low pK(a), it can penetrate rapidly into the cells and may inhibit vacuolar H(+) ATPase and prevent quinacrine-detectable vacuolar acidification without causing strong cell acidification. Since these drugs were found to penetrate into the cells, their lack of effect may reflect a higher resistance of both plasma membrane H(+)-ATPase and vacuolar ATPase to the drugs. Our data indicate that the lysosomotropic drugs under study have a dual action. On entering the cell, they cause intracellular acidification, very probably by inhibiting plasma membrane H(+)-ATPase and curtailing active proton pumping from the cells. Furthermore, they interfere with the function of V-type ATPase, causing vacuolar alkalinization and eventually cell death. PMID- 12655365 TI - Morphological and biochemical changes in human fibroblast lines induced by anthracyclines during apoptosis. AB - We show that treating human trisomic fibroblasts with anthracyclines - aclarubicin, daunorubicin and idarubicin - leads to certain changes in these cells; namely the activation of caspase 3, morphological changes and an increase in the level of intracellular calcium. These results suggest that anthracycline drugs are also able to induce apoptosis in pathological, trisomic cells. PMID- 12655366 TI - Carbamylation of proteins leads to alterations in the membrane structure of erythrocytes. AB - The effect of the sodium cyanate-induced carbamylation (carbamoylation) of proteins in erythrocytes was studied using spin labelling and spectrophotometric methods. The experiments were conducted in whole blood and in erythrocytes in phosphate buffer using 25 mmol/L of sodium cyanate. Lipid membrane fluidity was determined using three spin-labelled fatty acids: 5-, 12- and 16-doxylstearic acids (5-DS, 12-DS, 16-DS). Internal viscosity was measured with Tempamine, using also EPR spectroscopy. Osmotic fragility was determined spectrophotometrically. Incubation of whole blood with sodium cyanate led to an increase in lipid membrane fluidity in the deeper region of the lipid layer, indicated by 12- and 16-doxylstearic acid, and a decrease near the surface (5-DS). Statistically significant results were obtained for the internal viscosity and osmotic fragility of erythrocytes. An increase in internal viscosity and increase in osmotic fragility were found in erythrocytes after incubation of whole blood, as well as in erythrocytes incubated with sodium cyanate in buffer. Alterations in internal viscosity were stronger in erythrocytes incubated with sodium cyanate in blood than in erythrocytes in the buffer. On the other hand, higher osmotic fragility was observed for erythrocytes in the buffer. PMID- 12655367 TI - 52 kD Ro/SS-A localizes to punctate structures in the cytoplasm of epithelial cells. AB - Autoantibodies directed against 52 kD and 60 kD Ro/SS-A are frequently found in the sera of patients with lupus erythematosus and Sjogren's syndrome-related disorders. Their location in the cell is subject to continuous debate in literature. It has been postulated that 52 kD Ro (52 Ro) co-localizes with the 60 kD Ro autoantigen in the nucleus, while others demonstrated that 52 Ro is primarily cytoplasmic. In order to resolve this controversy, 52 Ro protein was tagged with green fluorescence protein, overexpressed in A431 keratynocytes, and its location determined using fluorescence confocal microscopy. The intracellular location of the fusion protein was revealed via GFP autofluorescene and indirect immunofluorescence microscopy, using purified anti-52 Ro antibodies. The cellular locations of native 52 Ro in normal human keratinocytes, and in human A431 keratinocyte and HepG2 hepatocyte cell lines were similarly determined by utilizing 2 human anti-52 Ro antibodies purified from two different non overlapping fragments of recombinant 52 Ro. In addition, colocalization of 52 Ro with mitochondria, lysosomes and endosomes was evaluated. It was found that both the 52 Ro-GFP fusion protein and the native 52 Ro localize in discrete cytoplasmic punctate structures separately from the mitochondria, lysosomes and endosomes. Furthermore, human autoantibodies that are reactive with denaturation sensitive epitopes on 52 Ro recognize these cytoplasmic punctate structures, whereas antibodies directed against denaturation-resistant 52 Ro epitopes do not. This explains why the previously used antibody against denaturation-resistant 52 Ro epitopes failed to detect the protein in such punctate structures. PMID- 12655368 TI - The influence of X-rays on human erythrocytes. Primary radicals. AB - The effects on human erythrocytes of water-derived radicals generated by X-rays were studied under anaerobic conditions and in the presence of oxygen. Erythrocyte damage was estimated on the basis of the reduced GSH and MetHb content in the erythrocytes, the -SH group content in the membrane proteins and the amount of K(+)released from the erythrocytes. The results obtained show that the level of reduced GSH was the most sensitive indicator of erythrocyte damage by X-rays followed by the efflux of K(+). The processes of GSH oxidation took place most rapidly under air. At a dose of 100 Gy, the level of GSH fell to about 50%, whereas under argon and N(2)O to about 75% and 65%, respectively. A slight increase in the efflux of K(+)was observed in preparations irradiated under air. However, when erythrocytes were irradiated under argon and N(2)O, the loss of K(+)occurred at a dose 8-times higher. Changes in the remaining parameters occurred at considerably higher doses. On the basis of the results obtained one can say that oxygen is a factor increasing the toxicity of(.)OH radicals towards erythrocytes; however, e(-)(aq)present in the system can cause a decrease in damage to certain cellular components. PMID- 12655369 TI - Is a fluid-mosaic model of biological membranes fully relevant? Studies on lipid organization in model and biological membranes. AB - The basic concept of the fluid-mosaic model of Singer and Nicolson, an essential point of which is that the membrane proteins are floating in a sea of excess lipid molecules organized in the lipid bilayer, may be misleading in understanding the movement of membrane components in biological membranes that show distinct domain structure. It seems that the lipid bilayer is an active factor in forming the membrane structure, and the lipid composition is responsible for the presence of domains in the membrane. The main role in the process of domain formation is played by cholesterol and sphingolipids. The results presented here show that in a binary mixture of cholesterol and unsaturated phospholipids, cholesterol is segregated out from the bulk unsaturated liquid-crystalline phase. This forms cholesterol-enriched domains or clustered cholesterol domains due to the lateral nonconformability between the rigid planar ring structure of cholesterol and the rigid bend of the unsaturated alkyl chain at double bond position. These cholesterol-enriched domains may be stabilized by the presence of saturated alkyl chains of sphingomyelin or glycosphingolipids, and also by specific proteins which selectively locate in these domains and stabilize them as a result of protein-protein interaction. Such lipid domains are called "rafts" and have been shown to be responsible both for signal transduction to and from the cell and for protein sorting. We also looked at whether polar carotenoids, compounds showing some similarities to cholesterol and affecting membrane properties in a similar way, would also promote domain formation and locate preferentially in one of the lipid phases. Our preliminary data show that in the presence of cholesterol, lutein (a polar carotenoid) may segregate out from saturated lipid regions (liquid-ordered phase) and accumulate in the regions rich in unsaturated phospholipids forming carotenoid-rich domains there. Conventional and pulse EPR (electron paramagnetic resonance) spin labeling techniques were employed to assess the molecular organization and dynamics of the raft-constituent molecules and of the raft itself in the membrane. PMID- 12655370 TI - Organization of antibiotic amphotericin B in model lipid membranes. A mini review. AB - Amphotericin B (AmB) is a polyene antibiotic frequently applied in the treatment of fungal infections. According to the general understanding, the mode of action of AmB is directly related to the molecular organization of the drug in the lipid environment, in particular to the formation of pore-like molecular aggregates. Electronic absorption and fluorescence techniques were applied to investigate formation of molecular aggregates of AmB in the lipid environment of liposomes and monomolecular layers formed at the argon-water interface. It appears that AmB dimers, stabilized by van der Waals interactions, are present in the membrane environment along with the aggregates formed by a greater number of molecules. Linear dichroism measurements reveal that AmB is distributed between two fractions of molecules, differently oriented with respect to the bilayer. Molecules in one fraction remain parallel to the plane of the membrane and molecules in the other one are perpendicular. Scanning Force Microscopy imaging of the surface topography of the monolayers formed with AmB in the presence of lipids reveals formation of pore-like structures characterized by the external diameter close to 17 A and the internal diameter close to 6 A. All the findings are discussed in terms of importance of the molecular organization of AmB in the pharmacological action, as well as of the toxic side effects of the drug. PMID- 12655371 TI - The nitroxides pirolin and pirolid protect the plasma membranes of rat cardiomyocytes against damage induced by anthracyclines. AB - This study was performed to evaluate the protective effects of pyrroline and pyrrolidine nitroxides Pirolin, PL, and Pirolid, PD, on the plasma membranes of rat cardiomyocytes treated in vitro with anthracycline drugs aclarubicin (ACL) and doxorubicin (DOX). The influence of two concentrations of drugs (10 and 20 microM) and nitroxides (0.1 and 1 mM) as well as their combinations (a drug and a nitroxide) on membrane fluidity was investigated. The plasma membranes of cardiomyocytes were labelled with a hydrophobic fluorescence probe 12-AS and membrane fluidity was estimated on the basis of the fluorescence anisotropy of the probe. We found that aclarubicin and doxorubicin induced a significant dose dependent decrease in membrane fluidity, whereas the nitroxides (PL and PD) caused its increase. Preincubation of cardiomyocytes with Pirolin entirely protected plasma membranes of these cells against damage caused by DOX. In the same conditions no protective effect of Pirolid was observed. What is more, Pirolid in combination with DOX caused fluidisation of the plasma membranes of cardiomyocytes. Both nitroxides at low concentration (0.1 mM) protected plasma membranes against rigidification induced by aclarubicin, while high concentration (1 mM) was ineffective and caused fluidisation of the plasma membranes of cardiomyocytes. PMID- 12655372 TI - Effects of pyrroline and pyrrolidine nitroxides on lipid peroxidation in heart tissue of rats treated with doxorubicin. AB - Protection from doxorubicin-induced lipid peroxidation in vivo by two pyrroline and pyrrolidine nitroxides, Pirolin, PL, and Pirolid, PD, was examined in the heart tissue of rats treated with this drug. The level of lipid peroxidation was estimated on the basis of MDA content. A considerable (three-fold) increase in the MDA amount was found in heart homogenates from rats injected with doxorubicin, whereas no significant changes in MDA content compared to control were observed in cardiomyocytes treated with the nitroxides (Pirolin or Pirolid) only. Pirolin injected simultaneously with doxorubicin showed antioxidative effect and markedly attenuated lipid peroxidation in the heart tissue caused by this drug. In contrast to Pirolin, structurally related Pirolid was ineffective in the protection of heart myocytes from DOX-induced lipid peroxidation. PMID- 12655373 TI - Testing cms-P-linked AFLPs for selection of rye hybrid components. AB - Application of AFLPs linked to pollen fertility restoration and non-performing genes evaluated in the C394-F2 hybrid was studied using a set of male sterile lines in the sterilising Pampa cytoplasm, several restorers and maintainer lines and, finally, two inbred lines backcrossed into cms-P, cms-R, cms-S and cms-C cytoplasms each. The set of male sterile lines based on the Pampa cytoplasm exhibited gradual variation in their ability to restore pollen fertility (starting from low and closing with high) in crosses with three unrelated restorers. Variations in the AFLPs between the analysed materials were observed, however, no clustering of the lines according to their sterile and fertile phenotypes was observed. The same markers, when applied to the population restorer (cv. Walet) that formed the C394-F2 cross permitted identification of plants with genotypes that could be recognized as restorers. PMID- 12655374 TI - Comprehensive analysis of all triple helical repeats in beta-spectrins reveals patterns of selective evolutionary conservation. AB - The spectrin superfamily (spectrin, alpha-actinin, utrophin and dystrophin) has in common a triple helical repeating unit of ~106 amino acid residues. In spectrin, alpha and beta chains contain multiple copies of this repeat. beta spectrin chains contain the majority of binding activities in spectrin and are essential for animal life. Canonical beta-spectrins have 17 repeats; beta-heavy spectrins have 30. Here, the repeats of five human beta-spectrins, plus beta spectrins from several other vertebrates and invertebrates, have been analysed. Repeats 1, 2, 14 and 17 in canonical beta are highly conserved between invertebrates and vertebrates, and repeat 8 in some isoforms. This is consistent with conservation of critical functions, since repeats 1, 2 and 17 bind alpha spectrin. Repeats 1 of beta-spectrins are not always detected by SMART or Pfam tools. A profile hidden Markov model of beta-spectrin repeat 1 detects alpha actinins, but not utrophin or dystrophin. Novel examples of repeat 1 were detected in the spectraplakins MACF1, BPAG1 and plectin close to the actin binding domain. Ankyrin binds to the C-terminal portion of repeat 14; the high conservation of this entire repeat may point to additional, undiscovered ligand binding activities. This analysis indicates that the basic triple helical repeat pattern was adapted early in the evolution of the spectrin superfamily to encompass essential binding activities, which characterise individual repeats in proteins extant today. PMID- 12655375 TI - The effect of hypochlorite on human erythrocytes pretreated with X-radiation. AB - Both hypochlorite and ionizing radiation induce oxidation processes of biomolecules. The effects are dependent to a large degree on the dose of the oxidizing agent. Previously we observed that split doses of gamma radiation caused lower hemolysis than the same but single doses. The critical factors influencing the occurrence of this effect were: the value of the first dose and the time between the doses. In this work we examined the effect of gamma radiation (40-400 Gy) on hemolysis of human erythrocytes induced by hypochlorite. Erythrocytes in PBS, hematocrit 2 %, were irradiated with doses of 40, 200 or 400 Gy. The dose-rate was 23.8 Gy/min. Cell suspensions were stirred during irradiation. After irradiation the erythrocytes were incubated for 1, 3 or 4 hours at room temperature and then hypochlorite was added to a 250 microM concentration. Control samples were erythrocytes treated only with NaOCl. The level of hemolysis was determined after NaOCl addition. Hemolysis of erythrocytes preirradiated with the dose of 400 Gy was lower than hemolysis of erythrocytes treated only with NaOCl. The effect was dependent on the time between the end of irradiation and the addition of NaOCl. In contrast, slightly higher hemolysis was observed for erythrocytes preirradiated with lower (40 or 200 Gy) doses of radiation. The observed effect is similar to that obtained for radiation-induced hemolysis. It suggests that ionizing radiation may induce structural and/or functional changes in erythrocytes, which make the cell more resistant to further oxidative damage. PMID- 12655376 TI - Cystocyte and lymphocyte derived fusomes/spectrosomes: analogies and differences: a mini-review. AB - Structures analogous to Drosophila spectrosomes were found in mammalian lymphocytes. Repasky and colleagues discovered an intracellular spectrin-rich structure in lymphoid cells, which had far-reaching parallels with the fusome/spectrosome of D. melanogaster germ cells. This fact implies that spectrosomes may be characteristic not only of insect germ cells, but also that an analogous structure may play an important role in other cell types. The term "spectrosome" was first used by Lin and Spradling in 1995 to describe a large sphere of fusomal material in D. melanogaster germline stem cells and their differentiated daughter cells - cytoblasts. In the D. melanogaster ovary, membrane skeletal proteins such as ankyrin, alpha/beta spectrin as well as adducin-like Hts protein(s) were found in this specific organelle - spectrosome/fusome. These orgalelles are involved in the creation of mitotic spindles and D. melanogaster cyst formation and oocyte differentiation, but the role of analogous spectrin-based aggregates found in nucleated cells still remains unclear. PMID- 12655378 TI - Induction and decay of thermotolerance in human erythrocytes determined by hemolysis. AB - Hemolysis was used as an endpoint for the measurement of damage to the plasma membrane in human erythrocytes after a single or a double heat shock. The thermotolerance of erythrocytes is a transitional phenomenon, reaching its maximum at a 3-hour incubation at 37 degrees C between the heat shocks. PMID- 12655377 TI - Resolving the ionotropic receptor kinetics and modulation in the time scale of synaptic transmission. AB - Synaptic transmission plays a crucial role in signal transduction in the adult central nervous system. It is known that synaptic transmission can be modulated by physiological and pathological processes and a number of factors including metal ions, pH, drugs, etc. The patch-clamp technique allows to measure postsynaptic currents, but the mechanism of these currents modulation remains unclear. The estimated value of neurotransmitter transient indicates that this time course is very short and the activation of postsynaptic receptors is extremely non-equilibrient. The ultrafast perfusion system makes it possible to mimic synaptic conditions and, additionally, the agonist concentration can be controlled, which is very important for pharmacokinetic studies. In the present paper, examples of pharmacological modulation of mIPSC kinetics and currents evoked by ultrafast agonist application are presented. PMID- 12655379 TI - [Evaluation of microsurgical treatment in a series of 121 intracranial aneurysms]. AB - OBJECTIVE: The results obtained with therapy of intracranial aneurysms, in terms of morbidity and mortality, are very important when the patient has to choose between microsurgical techniques or endovascular management. The aim of this paper is to review the information regarding current microsurgical treatment of intracranial aneurysms, and presenting our experience over the last five years. MATERIAL AND METHODS: We studied 101 consecutive patients with 121 intracranial aneurysms admitted between 1996 and 2000 with the initial diagnosis of subarachnoid hemorrhage. We paid special attention to the day of admission from the onset of the symptomatic hemorrhage to the grade of Hunt&Hess scale and the possibility of early or delayed microsurgical treatment. The diagnosis was based on four vessels cerebral angiography and in a few cases with CT-angiography. All patients were treated by microsurgical technique and such treatment was completed by nimodipine, intensive care unit management and in some cases of postoperative suspected vasospasm, induced arterial hypertension was applied. Post surgical angiography was carried out in all patients to confirm the clipping of the cerebral aneurysm. The 12 months assessment was based on the Glasgow Outcome Scale (GOS). RESULTS: The 92.1% of the patients were admitted with a grade equal or below III in the Hunt&Hess scale. A 80% were operated within the 72 hours of admission and in the remaining cases, the surgical treatment was delayed due to a grade IV or V or to a medical contraindication. Four patients died (3.9%). At 12 months follow up, 88.9% presented a score I or II in the GOS. CONCLUSION: According to our results, there are a substantial improvements in the microsurgical treatment of cerebral aneurysms, specially in patients admitted early after the onset of the symptoms of their hemorrhage, who have a grade I to III in the Hunt&Hess scale and showed a good level of consciousness. We think that the improvement of our results are due to: l. the high percentage of patients admitted with grades I to III. 2. the high percentage of patients operated within the first 72 hours from the onset of their symptomatic hemorrhage. 3. surgery was always carried out by the same two experienced vascular neurosurgeons. 4. intraoperative measures taken to prevent the rupture of the aneurysm. 5. early administration of nimodipine, ICU management, doppler studies and in seldom cases, induced hypertension therapy to treat the vasospasm and postoperative hypotension. PMID- 12655380 TI - [Giant supratentorial intracranial aneurysms. Analysis of 22 cases]. AB - INTRODUCTION: Giant intracranial aneurysms represent 2 to 5% of all aneurysms. They are well characterized from the anatomical and clinical point of view. Their natural history shows its potential lethality. Surgical treatment of giant aneurysms is a challenge for neurosurgeons. MATERIAL AND METHODS: Twenty-two patients were operated on through pterional craniotomy, specialized neuroanesthesia and microneurosurgical technics. Auxiliary methods like transitory clipping and retrograde decompression-suction technique were applied. Patients were followed at intensive care units and they were evaluated three months after the operation. Nineteen patients were in the fourth and sixth decade of life. Seventeen were females. Aneurysms were located at middle cerebral artery bifurcation; paraclinoidal carotid artery; proximal anterior cerebral artery and carotid bifurcation. Ninety one percent of aneurysms were clipped. Retrograde decompression-suction technique was performed in thirteen cases. RESULTS: Seventeen patients had good outcome and one patient died (4.5%). There were 6 postoperative complications and in four disappeared three months later. CONCLUSIONS: Giant aneurysms were operated on following main neurosurgical rules helped by auxiliar procedures to reduce aneurysms size and wall, aneurysms tension. New knowledge about giants aneurysms and the development of new techniques will permit better results. PMID- 12655382 TI - [Congenital brain tumors: nine cases and review of the literature]. AB - OBJECTIVE: To review congenital brain tumors patients younger than 2 months diagnosed in our Department. PATIENTS AND METHODS: Nine congenital brain tumors were diagnosed between 1983 and 2001 among a consecutive serie of 484 paediatric cerebral tumors (patients less than 15 years). Clinical and radiological findings and prognostic factors are analyzed. RESULTS: There were two teratomas, two glioblastomas, one xanthogranuloma affecting cavernous sinus, one supratentorial PNET and one medulloblastoma and one oligodendroglioma. In one case histological diagnosis was not possible. In four cases antenatal diagnosis was done by means of ultrasonography and MRI. Seven patients were operated on, with one intraoperative exitus. Five patients died due to tumoral progression in the first two months after surgery. Another case was considered not amenable for surgical resection, and thus was not operated. He died in the fifth day of life. Three patients are alive 14, 36 and 72 months after surgery, one with severe psicomotor delay, another one with normal mental status but dependent on seizures medication and the last one with mild mental delay. In the last case of our serie intrauterine death happened by 33(rd )week and histological diagnosis was not possible. CONCLUSIONS: Prognosis for congenital brain tumors depends on the operative morbidity and tumor histology. However final prognosis in these patients is still discouraging, despite early surgery and operative and anaesthetic improvements. PMID- 12655381 TI - Trigeminal nucleus caudalis dorsal root entry zone radiofrequency thermocoagulation for invalidating facial pain. AB - INTRODUCTION: Facial pain syndromes occasionally result in desperate clinical settings completely unresponsive to any known therapy. Trigeminal nucleus caudalis dorsal root entry zone (DREZ) lesion is reported to be of benefit in such cases. In 1982 Nashold performed the first DREZ caudalis lesion in a patient with anaesthesia dolorosa. PATIENTS AND METHODS: From 1994 to 2002 we have performed six DREZ caudalis lesions on five patients with extremely invalidating facial pain resistant to multiple pharmacological and surgical therapies. Pain was secondary to previous craniofacial surgery in all but one case. Pain presented as anaesthesia dolorosa or atypical facial pain so severe as to interfere with personal hygiene and even to prevent patients from oral feeding. A midline suboccipital approach was used and radiofrequency lesions (at the trigeminal nucleus caudalis in the cervicomedullary junction) were made at 1-mm intervals, 75 (o)C for 15 seconds each along the ipsilateral posterolateral sulcus from the cervical DREZ up to the obex. RESULTS: Pain relief was complete and permanent in two patients. Three patients experienced significant improvement but pain recurred in two (weeks to a few months after the procedure). No patient's pain was made worse. A patient with persistent postoperative nasolabial pain was re-operated on (improving again but ultimately remaining unchanged). Air venous embolism related to the sitting position (3 patients) during surgery and bradycardia due to manipulation in medulla (2 patients) occurred during some of the procedures without any cardiovascular or neurological repercussion. Postoperative complications included mild and transient ataxia and monoparesia (3 patients). DISCUSSION: Facial pain secondary to craniofacial surgery is known to be among the least responsive to treatment and a true challenge for pain clinicians. Actual indications for this procedure, operative technical details and the results of our series compared to previous reports are reviewed. CONCLUSION: Trigeminal nucleus caudalis radiofrequency thermocoagulation is an effective neurosurgical procedure for the treatment of chronically debilitating and desperate facial pain syndromes with acceptable morbidity. PMID- 12655383 TI - [Non invasive intracranial hyperthermia with Electric Capacitive Transference ECT- Intratumoral and cerebral thermometry results]. AB - The aim of this work is to present the results of thermic increase obtained at the brain and intratumoral levels through a non invasive technique -Electric Capacitive Transference- (ECT), developed in 1985 by Indiba. A review of the literature does not provide any reference of cerebral and intratumoral thermometry in real time with a non invasive technique of intracranial hyperthermia. In the 8 studied patients, the increases of temperature in the brain, ranged between 0.7 and 1.5 degrees C in relation to the depth of the thermometric probe and the incidence angle of the external electrode. Between tumoral and perilesional brain tissues, thermic increase was 0.3 to 0.7 degrees C greater at tumoral level. The observation that in no case the surrounding brain tissue registered a temperature over 39.2 degrees C supports the harmlessness of the technique regarding the potential damage to healthy brain tissue and seems to confirm previous data obtained in anatomopathological studies in animal experimentation performed in 1990, which showed an absence of lesions in tissues and organs. The greater and somewhat more prolonged thermic increase observed at tumoral level has been called the "greenhouse effect". PMID- 12655384 TI - [Oligoastrocytoma with signet-ring cell differentiation. A morphological, ultrastructural and immunohistochemical study]. AB - We present a case of a mixed glial tumor (oligoastrocytoma) with signet-ring cells. This cellular feature is a rare differentiation in glial tumors of the central nervous system. Histological, immunohistochemical and ultrastructural findings have been analyzed. Signet-ring cells showed intense expression with GFAP, S-100 and vimentin. A differential diagnosis with other primary brain tumors and cerebral metastases with signet-ring cell differentiation was discussed. PMID- 12655385 TI - [Cauda syndrome due to spinal stenosis and diskytis in two different spinal levels. Diagnosis using myelo-TC]. AB - We report a case of a 75 year old man presenting with paraparesis, urinary incontinence and saddle anesthesia evolving after several months and severe lumbalgia and bad general status which developed fewdays before admission. CT scan showed spondilosis with lytic lesion at L5-S1 level. MR1 was not performed because the patient had a pacemaker. A myelo-CT study was performed showing a complete stop at L4-L5 level together with L5-S1 diskytis. Laminectomy of L5 and discectomy L5-S1 was performed obtaining tissue sample diagnostic of diskytis and osteomyelitis. Our patient had two lesions at two different levels with two independent syndromic appearence. Myelo-CT played an important diagnostic role in this case because MRI study could not be obtained. PMID- 12655386 TI - [Differential diagnosis of multiple metastasis]. PMID- 12655388 TI - [Tale: till the next congress]. PMID- 12655389 TI - Outcomes of patients referred for confirmation of brain death by 99mTc exametazime scintigraphy. AB - OBJECTIVE: To review clinical outcomes in patients referred for confirmation of brain death by (99m)Tc-labelled exametazime scintigraphy and to assess any tangible benefit. DESIGN AND SETTING: A retrospective casenote review in the intensive care unit and nuclear medicine department of a tertiary-level acute care hospital. PATIENTS AND PARTICIPANTS: 66 consecutive patients, in whom a clinical diagnosis of brain death was not possible, undergoing (99m)Tc-labelled exametazime scintigraphy from February 1993 to March 2002, and for 56 of whom casenotes were available for review. MEASUREMENTS AND RESULTS: Brain death was confirmed in 77% on the basis of absent supratentorial and infratentorial uptake. Where brain death was not confirmed, a normal scan was seen in 30%, the remainder showing patchy reduced infratentorial and/or supratentorial uptake. All patients with the scintigraphic appearance of brain death subsequently died. Even when brain death could not be confirmed, 11 of 13 patients died soon afterwards. Two patients with normal brain scintigraphy regained consciousness and survived for an extended period. When brain death was not confirmed by scintigraphy, elapsed time to certification of death was longer (44.7 vs. 7 h) and organ donation was less common even when a subsequent clinical diagnosis was made. CONCLUSIONS: The results indicate that patient prognosis remains poor regardless of the findings on (99m)Tc-labelled exametazime scintigraphy, but that the scan result influences the elapsed time to withdrawal of therapy or organ donation, providing a significant cost benefit, and occasionally a long-term survivor is seen. PMID- 12655391 TI - Comment on "Terlipressin in chronic hyperdynamic endotoxic shock: is it safe?" by P. Asfar. PMID- 12655394 TI - Thoughts on intensive care: from morphine to Merlyn and Tolstoy to television. PMID- 12655393 TI - High levels of sulfated mucins in bronchoalveolar lavage fluid of ICU patients with ventilator-associated pneumonia. AB - OBJECTIVE: To compare the levels of sulfated mucins in bronchoalveolar lavage fluid (BALF) in ICU patients with ventilator-associated pneumonia (VAP) with those in non-infectious controls, i.e., ventilated ICU patients without VAP, and nonventilated patients. DESIGN AND SETTING: Prospective study in a mixed intensive care unit and outpatient clinic of a university hospital. PATIENTS: The study included 56 ventilated ICU patients with VAP, 21 ventilated ICU patients without VAP, and 26 nonventilated outpatients with no pulmonary infection. MEASUREMENTS AND RESULTS: Total cell count and differential cell count of BALF samples were determined, and sulfated mucin levels were measured. For this we used the monoclonal antibody F2 against the sulfated Lewis C structure (SO(3) 3Galbeta1-3GlcNAc). Sulfated mucin levels were significantly increased in ICU patients with VAP than in those without VAP and nonventilated patients. No statistical difference was found between the two groups of ICU patients regarding APACHE II score and the duration of mechanical ventilation prior to the bronchoalveolar lavage. CONCLUSIONS: The increased levels of sulfated mucins in ICU patients with VAP are associated with infection and not with ventilation. The increase in sulfated mucins may favor the persistence of those micro-organisms that possess mucin sulfatase activity, which enable them to survive within the mucus, especially Pseudomonas aeruginosa. PMID- 12655390 TI - Cerebral energy metabolism during transient hyperglycemia in patients with severe brain trauma. AB - OBJECTIVE: To study whether transient hyperglycemia adversely affects cerebral energy metabolism in patients with severe traumatic brain lesions. DESIGN AND SETTING: Prospective, nonrandomized study in the neurosurgical intensive care unit of a university hospital. PATIENTS: 108 patients treated for severe traumatic brain lesions. INTERVENTIONS: All patients were treated according to neurosurgical intensive care routine including monitoring of intracranial pressure. One microdialysis catheter was inserted via a burr hole frontally to that used for the intraventricular catheter ("better" position). In patients with focal lesions one or more catheters were inserted into cerebral cortex surrounding an evacuated focal contusion or underlying an evacuated hematoma ("worse" position). Perfusion rate was 0.3 micro l/min and samples were taken every 30 or 60 min. The levels of glucose, pyruvate, lactate, glutamate, and glycerol were analyzed and displayed bedside. MEASUREMENTS AND RESULTS: There were 18 episodes of moderate (12-15 mmol/l) and 6 episodes of pronounced (>15 mmol/l) hyperglycemia. Moderate hyperglycemia did not change intracerebral levels of lactate, pyruvate, glutamate, glycerol, or lactate/pyruvate ratio. Lactate concentrations increased during pronounced hyperglycemia. Pronounced cerebral lactic acidosis and a moderate increase in interstitial glycerol concentration indicating cell membrane degradation was observed in a single patient with pronounced, long-lasting hyperglycemia. CONCLUSIONS: Cerebral energy metabolism was affected by transient hyperglycemia only at blood glucose concentration above 15 mmol/l as shown by a moderate increase in interstitial lactate level. PMID- 12655395 TI - A computer program for interpreting pulmonary artery catheterization data: results of the European HEMODYN Resident Study. AB - We previously validated an expert computer program (Hemodyn) designed to assist in interpreting pulmonary artery catheterization data. The present multicentric study assessed the influence of Hemodyn on the therapeutic strategies of residents. Each resident made several diagnostic choices and suggested appropriate treatments based on pulmonary artery catheterization (PAC) data. After knowledge of the computer interpretation, the resident could either maintain or change his or her diagnosis and treatment under a senior supervision. Agreement between the residents' initial evaluation and the computer's was poor (kappa <0.6). After computer assistance, agreement improved dramatically (kappa >0.9). Computer assistance led the residents to change at least one suggested treatment in 63% of cases, and in 8% of cases the residents changed the initial suggestion to its opposite. Expert software capable of helping residents to interpret PAC data properly may improve the quality of care given to critically ill patients. PMID- 12655396 TI - Effects of selective inactivation of individual genes for low-molecular-mass subunits on the assembly of photosystem II, as revealed by chloroplast transformation: the psbEFLJoperon in Nicotiana tabacum. AB - Photosystem (PSII) is a supramolecular polypeptide complex found in oxygenic photosynthetic membranes, which is capable of extracting electrons from water for the reduction of plastoquinone. An intriguing feature of this assembly is the fact that it includes more than a dozen low-mass polypeptides of generally unknown function. Using a transplastomic approach, we have individually disrupted the genes of the psbEFLJoperon in Nicotiana tabacum, which encode four such polypeptides, without impairing expression of downstream loci of the operon. All four mutants exhibited distinct phenotypes; none of them was capable of photoautotrophic growth. All mutants bleached rapidly in the light. Disruption of psbEand psbF, which code for the alpha and beta apoproteins of cytochrome b(559), abolished PSII activity, as expected; Delta psbL and Delta psbJ plants displayed residual PSII activity in young leaves. Controlled partial solubilisation of thylakoid membranes uncovered surprisingly severe impairment of PSII structure, with subunit and assembly patterns varying depending on the mutant considered. In the Delta psbL mutant PSII was assembled primarily in a monomeric form, the homodimeric form was preponderant in Delta psbJ, and, unlike the case in Delta psbZ, the thylakoids of both mutants released some PSII supercomplexes. On the other hand, Photosystem I (PSI), the cytochrome b(6)f complex, ATP synthase, LHCII, and CP24/CP26/CP29 antennae were present in near wild-type levels. The data are discussed in terms of their implications for structural, biogenetic and functional aspects of PSII. PMID- 12655397 TI - The Aspergillus parasiticus protein AFLJ interacts with the aflatoxin pathway specific regulator AFLR. AB - The Aspergillus parasiticus aflJgene, which is located in the aflatoxin biosynthetic gene cluster and is transcribed divergently from the aflatoxin pathway regulatory gene aflR, encodes a 438-amino acid protein. Transformation with aflJplus aflR, but not aflJalone, increased the accumulation of aflatoxin precursors substantially in an O-methylsterigmatocystin-accumulating A. parasiticus strain. Disruption of aflJ resulted in non-pigmented mutants that lost the ability to synthesize aflatoxin intermediates. Transcript profiling by real time RT-PCR indicated that the lack of aflJtranscripts in the aflJ knockout mutants is associated with a significant decrease in the transcript levels of the genes for early (pksAand nor1), middle (ver1) and later (omtA) steps in the aflatoxin biosynthetic pathway, with the degree of reduction ranging from 5- to 20-fold. Deletion of aflJ, however, did not have any effect on the aflRtranscript level, and vice versa. Two-hybrid assays showed that AFLJ did not interact with aflatoxin biosynthetic enzymes, including NOR1, VER1, OMTA and ORDA. But AFLJ interacted with full-length AFLR, and the DNA-binding domain of AFLR was found not to be essential for this interaction. Simultaneous substitution of Arg427, Arg429, and Arg431 in the C-terminal region of AFLR with Leu residues abolished its ability to interact with AFLJ. Replacement of Asp436, which was previously shown to be crucial for AFLR's activation activity, with His, in contrast, had little effect on the interaction. On the other hand, deletions in most regions of AFLJ appeared to destroy its function, despite the fact that random amino acid substitution(s) in its C-terminal region did not drastically affect its capacity to interact with AFLR. The results show that aflJis involved in the expression of aflatoxin structural genes and support the hypothesis that aflJis a coactivator gene. PMID- 12655398 TI - Construction of a BAC library and generation of BAC end sequence-tagged connectors for genome sequencing of the African malaria mosquito Anopheles gambiae. AB - A Bacterial Artificial Chromosome (BAC) genomic DNA library of Anopheles gambiae, the major human malaria vector in sub-Saharan Africa, was constructed and characterized. This library (ND-TAM) is composed of 30,720 BAC clones in eighty 384-well plates. The estimated average insert size of the library is 133 kb, with an overall genome coverage of approximately 14-fold. The ends of approximately two-thirds of the clones in the library were sequenced, yielding 32,340 pair-mate ends. A statistical analysis (G-test) of the results of PCR screening of the library indicated a random distribution of BACs in the genome, although one gap encompassing the white locus on the X-chromosome was identified. Furthermore, combined with another previously constructed BAC library (ND-1), ~2,000 BACs have been physically mapped by polytene chromosomal in situ hybridization. These BAC end pair mates and physically mapped BACs have been useful for both the assembly of a fully sequenced A. gambiae genome and for linking the assembled sequence to the three polytene chromosomes. This ND-TAM library is now publicly available at both http://www.malaria.mr4.org/mr4pages/index.html/ and http://hbz.tamu.edu/, providing a valuable resource to the mosquito research community. PMID- 12655400 TI - Instability of short-sequence DNA repeats of pear pathogenic Erwinia strains from Japan and Erwinia amylovora fruit tree and raspberry strains. AB - An array of short-sequence DNA repeats (SSRs) occurs in the plasmid pEA29 of the fire blight pathogen Erwinia amylovora. A large number of "fruit tree" strains, mainly from Central and Western Europe, were screened for their SSR numbers, and the analyses were extended to five raspberry strains from North America and six pear pathogenic Erwinia strains from Japan. The repeat ATTACAGA present in all E. amylovorastrains was found to be reiterated 3 to 15 times. The Japanese strains contained the major repeat sequence GGATTCTG, which was reiterated 16 to 24 times. ATTACAGG, which resembles the SSR of E. amylovora, was reiterated two or three times. In a novel approach, sequencing gels were used to visualize the rare occurrence of shorter arrays (down to three repeats) in E. amylovoraand the Japanese Erwinia strains. Changes in the repeat numbers in E. amylovora were observed repeatedly when the bacteria had been exposed to stress conditions. The repeat structures of homo- and heteroduplices of PCR-amplified repeats were also analyzed by cleavage of annealed molecules with the single-strand-specific endonuclease from bacteriophage T4. Not only heteroduplexes, but also homoduplexes showed non-matching regions in the SSRs, which could arise from transient formation of loops due to strand slippage during the assays. PMID- 12655399 TI - Mutant telomeres inhibit transcriptional silencing at native telomeres of the yeast Kluyveromyces lactis. AB - We report the identification and characterization of transcriptional silencing at native telomeres in the budding yeast Kluyveromyces lactis. We show that K. lactis telomeres are able to repress the transcription of a gene located at the junction between the telomeric repeat tract and the subtelomeric domain. As in Saccharomyces cerevisiae, switching between the repressed and derepressed transcriptional states occurs. C-terminal truncation of the telomere binding protein Rap1p, which leads to a regulated alteration in telomere length, reduces telomeric silencing. In addition, telomeric silencing is reduced dramatically in telomerase RNA mutants in which telomere length control has been lost. This is consistent with the possibility that the structure of the entire telomere affects the silencing functions exhibited by its internal domain. PMID- 12655401 TI - Meiotic segregation of a homeologous chromosome pair. AB - During meiosis, the alignment of homologous chromosomes facilitates their subsequent migration away from one another to opposite spindle poles at anaphase I. Recombination is part of the mechanism by which chromosomes identify their homologous partners, and serves to link the homologs in a way that, in some organisms, has been shown to promote proper attachment to the meiotic spindle. We have built a diploid strain that contains a pair of homeologous chromosomes V': one is derived from Saccharomyces cerevisiae and one originates from S. carlsbergensis. Sequence analysis reveals that these chromosomes share 71% sequence identity. The homeologs experience high levels of meiotic double stranded breaks. Despite their relatedness and their competence to initiate recombination, the meiotic segregation behavior of the homeologous chromosomes suggests that, in most meioses, they are partitioned by a meiotic segregation system that has been shown previously to partition non-exchange chromosomes and pairs with no homology. Though the homeologous chromosomes show a degree of meiotic segregation fidelity similar to that of other non-exchange pairs, our data provide evidence that their limited sequence homology may provide some bias in meiotic partner choice. PMID- 12655403 TI - Tourist C transposable elements are closely associated with genes expressed in flowers of rice (Oryza sativa). AB - Tourist elements comprise a group of transposable elements in plants. One of these elements, Tourist-OsaCatA(a Tourist C element), has been found in the 5; flanking region of a catalase gene, CatA, in rice (Oryza sativa). Using reverse transcriptase-PCR (RT-PCR) analyses of leaves, roots, flowers and developing seeds of rice, we assessed the transcription levels of ten known genes containing Tourist C elements, and of three additional putative genes for which expressed sequence tags (ESTs) including Tourist C elements have been isolated. We found that nine of the ten known genes and two of the three represented by ESTs were expressed in at least one of the organs we analyzed, and all of the genes detected were expressed in flowers, usually in stamens or pistils. We also assessed the expression of the 29 Tourist C-containing hypothetical coding sequences (CDSs) obtained so far by high-throughput genomic sequencing. We found that CDSs of all 11 genes whose transcripts were detectable by RT-PCR were expressed in flowers, especially in stamens or pistils. In contrast, RT-PCR analyses of genes or CDSs associated with other miniature inverted-repeat transposable elements (MITEs), such as Tourist D, Gaijin, Explorer, and Castaway, showed that some of them were expressed only minimally or not at all in flowers. Therefore, compared with other MITEs, Tourist C elements seem to show a strong association with genes that are expressed in the flowers of rice. PMID- 12655402 TI - The transmembrane domain of the DnaJ-like protein DjlA is a dimerisation domain. AB - DjlA is a bitopic inner membrane protein, which belongs to the DnaJ co-chaperone family in Escherichia coli. Overproduction of DjlA leads to the synthesis of colanic acid, resulting in mucoidy, via the activation of the two-component regulatory system RcsC/B that controls the cps (capsular polysaccharide) operon. This induction requires both the co-chaperone activity of DjlA, in cooperation with DnaK and GrpE, and its unique transmembrane (TM) domain. Here, we show that the TM segment of DjlA acts as a dimerisation domain: when fused to the N terminal DNA-binding domain of the lambda cI repressor protein, it can substitute for the native C-terminal dimerisation domain of cI, thus generating an active cI repressor. Replacing the TM domain of DjlA by other TM domains, with or without dimerising capacity, revealed that dimerisation is not sufficient for the induction of cps expression, indicating an additional sequence- or structurally specific role for the TM domain. Finally, the conserved glycines present in the TM domain of DjlA are essential for the induction of mucoidy, but not for dimerisation. PMID- 12655404 TI - Patterns of Hermes transposition in Drosophila melanogaster. AB - Transposable elements are being developed as tools for genomics and for the manipulation of insect genotypes for the purposes of biological control. An understanding of their transposition behavior will facilitate the use of these elements. The behavior of an autonomous Hermes transposable element from Musca domestica in the soma and germ-line of Drosophila melanogaster was investigated using the method of transposon display. In the germ-line, Hermes transposed at a rate of approximately 0.03 jumps per element per generation. Within the soma Hermes exhibited markedly non-random patterns of integration. Certain regions of the genome were distinctly preferred over others as integration targets, while other regions were underrepresented among the integration sites used. One particular site accounted for 4.4% of the transpositions recovered in this experiment, all of which were located within a 2.5-kb region of the actin5C promoter. This region was also present within the Hermes element itself, suggesting that this clustering is an example of transposable element "homing". Clusters of integration sites were also observed near the original donor sites; these represent examples of local hopping. The information content (sequence specificity) of the 8-bp target site was low, and the consensus target site resembles that determined from plasmid-based integration assays. PMID- 12655405 TI - The different (sur)faces of Rap1p. AB - The DNA-binding protein Rap1p fulfills many different functions in the yeast cell. It targets 5% of the promoters, acting both as a transcriptional activator and as a repressor, depending on the DNA sequence context. In addition, Rap1p is an essential structural component of yeast telomeres, where it contributes to telomeric silencing. Here we review the evidence indicating that Rap1p function is modulated by the precise architecture of the its binding site and its surroundings: long tracts of telomeric repeats for telomeric functions, specific sequences and orientation for maximal transcriptional activation, and specific DNA recognition sequences for complementary factors in other cases. Many of these functions are probably related to chromatin organization around Rap1p DNA binding sites, resulting from the very tight binding of Rap1p to DNA. We propose that Rap1p alters its structure to bind to different versions of its DNA binding sequence. These structural changes may modulate the function of Rap1p domains, providing different interacting surfaces for binding to specific co-operating factors, and thus contributing to the diversity of Rap1p function. PMID- 12655406 TI - Structure and expression of spermidine synthase genes in apple: two cDNAs are spatially and developmentally regulated through alternative splicing. AB - Three cDNAs (MdSPDS1, 2a and 2b) encoding spermidine synthase (SPDS), a key enzyme in the polyamine biosynthesis, have been cloned from apple [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.]. The deduced amino acid sequences of their protein products share 76-83% identity with SPDSs of other higher plants. A comparison of the sequences of the three cDNAs and of the two corresponding genomic DNA fragments (SPDS1 and SPDS2) indicated that MdSPDS1 was transcribed from the SPDS1 sequence, whereas MdSPDS2a and MdSPDS2b were both derived from SPDS2 by alternative splicing. To learn more about the physiological roles of MdSPDS1, MdSPDS2a and MdSPDS2b, Northern analyses were carried out, together with measurements of polyamine content. Levels of both MdSPDS1 and MdSPD2a were higher in young leaves than in mature leaves and shoots. In fruits, mRNA levels were nearly as high as in young leaves and remained high during fruit development. By RT-PCR, MdSPDS2b transcripts were detected in mature leaves and shoots, but not in young leaves and fruits. These results indicate that MdSPDS2a and MdSPDS2b are differentially regulated in a tissue- and developmentally specific manner. The content of free polyamines in mesocarp tissues was measured at five stages of fruit development. At all stages, spermidine (Spd) was the predominant form of polyamine. The level of Spd was high at the early growth stage and declined to about 90% during later developmental stages. The possible regulation of SPDS expression during apple fruit development is discussed. PMID- 12655408 TI - Characterization of the ugpG gene encoding a UDP-glucose pyrophosphorylase from the gellan gum producer Sphingomonas paucimobilis ATCC 31461. AB - The ugpGgene, which codes for a UDP-glucose pyrophosphorylase (UGP) (or glucose-1 phosphate uridylyltransferase; EC 2.7.7.9) in Sphingomonas paucimobilis ATCC 31461, was cloned and sequenced. This industrial strain produces the exopolysaccharide gellan, a new commercial gelling agent, and the ugpG gene may convert glucose-1-phosphate into UDP-glucose in the gellan biosynthetic pathway. The ugpG gene is capable of restoring the capacity of an Escherichia coli galU mutant to grow on galactose by functional complementation of its deficiency for UDP-glucose pyrophosphorylase activity. As expected, the predicted gene product shows strong homology to UDP-glucose pyrophosphorylases from several bacterial species. The N-terminal region of UgpG exhibits the motif GXGTRXLPXTK, which is highly conserved among bacterial XDP-sugar pyrophosphorylases, and a lysine residue (K(192)) is located within a VEKP motif predicted to be essential for substrate binding or catalysis. UgpG was purified to homogeneity as a heterologous fusion protein from crude cell extracts prepared from IPTG-induced cells of E. coli, using affinity chromatography. Under denaturing conditions, the fusion protein S-UgpG-His(6) migrated with an estimated molecular mass of 36 kDa [corresponding to the predicted molecular mass of native UgpG (31.2 kDa) plus 5 kDa for the S and histidine tags). Kinetic analysis of UgpG in the reverse reaction (pyrophosphorolysis) showed a typical Michaelis-Menten substrate saturation pattern. The apparent K(m) and V(max) values estimated for UDP-glucose were 7.5 microM and 1275 micromol/min/g. PMID- 12655407 TI - Interfering mutations provide in vivo evidence that Escherichia coli SecE functions in multimeric states. AB - SecY, SecE and SecG form a heterotrimer, which functions as a protein translocation channel in Escherichia coli. The cytosolic loop of SecE contains a segment that is conserved among different organisms. Here we show that mutational alterations in this segment not only inactivate the SecE function but confer dominant interfering properties on the altered SecE molecule. Such effects were especially evident in mutant cells in which the requirement for SecE function was increased. Overproduction of SecE, but not of SecY, alleviated the dominant negative effects. These results suggest that the inactive SecE molecule sequesters wild-type SecE. It was also found that an amino acid substitution, D112P, in the C-terminal periplasmic region intragenically suppressed the dominant interference. These results are consistent with a notion that there is significant SecE-SecE interaction in vivo, in which the C-terminal region has an important role. The data hence suggest that dimeric SecE participates in the formation of the functional translocation channel. PMID- 12655409 TI - Mutations in GCR1 affect SUC2 gene expression in Saccharomyces cerevisiae. AB - Transcription of SUC2, the gene that encodes the cytoplasmic and secreted forms of the enzyme invertase, is controlled by glucose repression and derepression mechanisms in Saccharomyces cerevisiae. Several regulatory factors such as the Mig1p-Tup1p-Ssn6p repressor complex and the Snf1p kinase complex have been identified previously as regulators of SUC2 expression. We show that, in addition to these factors, expression of SUC2 is affected by mutations in the gene GCR1 that encodes the glycolysis regulatory protein Gcr1p. Expression of Suc2-LacZ was not repressed by glucose in gcr1 mutant yeast cells exposed to glucose. Furthermore, secreted invertase activity was constitutively expressed under glucose-repressed and derepressed conditions in gcr1 mutants. DNA gel mobility shift assays and in-vitro DNase I protection experiments mapped a DNA binding site for Gcr1p in the transcriptional control region of the SUC2 gene, next to a previously mapped Mig1p binding site. However, the mechanism by which gcr1 mutations relieve glucose repression remains obscure. PMID- 12655411 TI - Post-dural puncture headaches in children. A literature review. AB - Post-dural puncture headache (PDPH) is a well recognised complication of spinal and epidural anaesthesia. It can also occur after diagnostic or therapeutic lumbar puncture. Few cases have been reported in children. We reviewed the literature regarding definition, aetiology, incidence, risk factors, prevention and treatment, in order to provide some recommendations. Significant factors include age, gender, needle diameter, needle tip design, orientation of the tip during puncture, previous PDPH, history of migraine and repeated attempts to achieve puncture. There is no evidence for the use of increased fluids or bed rest to prevent PDPH. Once the diagnosis is made, conservative therapy is recommended for 48 h. Persistent PDPH can be treated in several ways; an epidural blood patch is one of the most effective methods. CONCLUSION: occurrence of post dural puncture headache after lumbar puncture in children is rare. There are conflicting data about risk factors, prevention and treatment. PMID- 12655412 TI - Immunonutrients and neonates. AB - The gastrointestinal tract is the largest surface area of the body and the primary site for microorganisms, foreign antigens and toxins to gain entry to the host's internal milieu. The use of enteral feedings enriched with immune enhancing ingredients is attracting considerable interest because there is increasing application of enteral feeding and appreciation of the role of the gut in the development of infection and of multiple organ failure in critically ill patients. CONCLUSION: in this review, we will discuss nutrients, such as glutamine, arginine, omega 3 fatty acids, nucleotides, probiotics, and lactoferrin, and how they might be used as immunonutrients in neonatal clinics. PMID- 12655413 TI - Unusual presentation of three siblings with familial heterozygous hypobetalipoproteinaemia. AB - We describe three siblings with the unusual presentation of manifest steatorrhoea and vitamin E deficiency mimicking homozygous familial hypobetalipoproteinaemia (FHBL) but whose lipid profile (cholesterol and ApoB) was consistent with heterozygous FHBL. Upper gastrointestinal endoscopy and small intestinal biopsy were normal. We discuss the diagnosis with reference to the relevant literature. CONCLUSION: although rare, familial hypobetalipoproteinaemia should be considered among the causes of manifest steatorrhoea in childhood even without evidence of failure to thrive. Dietary restriction of fat and high dose vitamin E supplementation improves quality of life by reducing stool frequency and may prevent or delay neurological complications. PMID- 12655414 TI - Two cases with transient lipoprotein lipase (LPL) activity impairment: evidence for the possible involvement of an LPL inhibitor. AB - Two independent severe hypertriglyceridemic infants with transiently impaired lipoprotein lipase (LPL) activity were observed and the causes were explored. Both infants were female, born prematurely with low birth weight and developed hypertriglyceridemia (Fredrickson type V hyperlipidemia: high VLDL and low LDL/HDL) a few months after birth. While mass levels of their post-heparin plasma LPL and apoprotein C-II (apo C-II), a physiological activator of LPL, were normal, their post-heparin plasma LPL activities were remarkably impaired. Both of their mothers' post-heparin plasma LPL activities were slightly or moderately impaired as well, without a decrease in the LPL mass level. No mutations in the genes for LPL and apo C-II were detected in either patient. In an in vitro study with their serum at onset, we could not detect any distinct circulating inhibitors for LPL. There was no data supporting infection or autoimmune diseases, which might have an impact on LPL activity, during the follow-up period. Levels of their plasma triglyceride (TG) and total cholesterol (TC) were decreased quickly by a dietary intervention with medium-chain triglyceride (MCT) milk and kept normal even after stopping the intervention at around age 1 year. However, their low post-heparin LPL activity persisted and returned to normal at around age 2 years. Their low HDL cholesterol levels persisted even after recovery of the TG and TC levels, although lecithin:cholesterol acyltransferase (LCAT) and cholesterol-ester-transfer protein (CETP), two key enzymes of HDL metabolism, were normal throughout the course. The exact reasons why their post heparin LPL activities were impaired for a certain period and why their HDL cholesterol levels have remained low are still unclear. CONCLUSION: Transiently impaired LPL activity with no defect in LPL enzyme induced severe hypertriglyceridemia in infants. The transient occurrence of inhibitor(s) for LPL was proposed. PMID- 12655415 TI - Insulin sensitivity and beta-cell secretion in thalassaemia major with secondary haemochromatosis: assessment by oral glucose tolerance test. AB - Diabetes mellitus in patients with thalassaemia major is caused by secondary haemochromatosis due to transfusional iron overload. The pathogenetic mechanisms leading from siderosis to diabetes are still poorly understood. This study aimed at assessing the influence of insulin resistance and insulin deficiency on that process. Glucose, insulin and C-peptide levels during oral glucose tolerance tests (OGTT) from 36 thalassaemic patients with normal ( n=23), impaired ( n=6), or diabetic glucose tolerance ( n=7) and 32 control subjects were examined. Insulin secretion and insulin sensitivity were assessed by established calculated indices. Fasting, 2h and integrated glucose concentration were significantly increased in thalassaemic patients with normal glucose tolerance compared to controls (5.01/4.59 mmol/l, 6.33/5.17 mmol/l, and 844.2/739.3 mmol/l per min, respectively; all P<0.03). Patients with impaired glucose tolerance presented hyperinsulinaemia and delayed peak insulin during OGTT. The C-peptide/insulin ratio was decreased in patients with abnormal glucose tolerance compared to controls (5.85/7.33 x 10(3)pmol/l per min, P<0.03). It was negatively correlated with age in patients ( r=-0.45, P<0.01), but positively in controls ( r=0.43, P<0.03). Insulin sensitivity was significantly reduced in patients with impaired glucose tolerance or diabetes compared to controls. In addition, a significant decrease in patients with normal glucose tolerance was shown by two insulin sensitivity indices (all P<0.05). In thalassaemia patients, insulin sensitivity was negatively correlated with age. Insulin secretion capacity according to the homeostasis assessment model was significantly reduced in patient groups compared to controls (Kruskal-Wallis-test, P<0.004). CONCLUSION: Insulin resistance is of central importance for the development of diabetes mellitus in patients with secondary haemochromatosis. An additional early defect in beta-cell secretion cannot be excluded. PMID- 12655416 TI - Transmesenteric hernia: report of two patients with diagnostic emphasis on plain abdominal X-ray findings. AB - Transmesenteric hernia is a rare cause of small bowel obstruction and is seldom diagnosed preoperatively, partly because of unfamiliarity with this type of internal hernia. The clinical symptoms of internal hernia may be intermittent and nonspecific, making the diagnosis extremely difficult. We report two patients, 7- and 5-year-old girls, with mesenteric hernia of the ileum. Because of the difficulty of clinical diagnosis of internal hernia, imaging studies played a crucial role. We would like to stress the importance of plain radiological findings as diagnostic aids. CONCLUSION: a consistent intestinal gas imaging after some interval suggests the possibility of an internal hernia, especially accompanied with a circular or oval defect of the gas shadows in the middle of the abdomen. A serial abdominal X-ray study can be helpful in the diagnosis of internal hernia. PMID- 12655417 TI - Clinical data and cerebrospinal fluid findings in Lyme meningitis versus aseptic meningitis. AB - The purpose of this study was to characterise Lyme meningitis (LM) in a Belgian paediatric population and to suggest findings that could allow early distinction from aseptic meningitis (AM). The medical records of patients hospitalised between 1993 and 2000 and with a discharge diagnosis of LM (n=14) or AM (n=16) were retrospectively reviewed. The Kruskal-Wallis test was used to compare data. Of the patients, 16 were identified with AM and 14 with LM, amongst which 7 presented with isolated LM and 7 with associated peripheral facial palsy (PFP). Patients with AM, when compared with LM, complained of more pronounced signs and symptoms of meningitis (fever, headaches, and neck stiffness); they statistically displayed a shorter period of symptoms before admission (1.6 vs 15 days), higher neutrophilic component (mean 56% vs 2.4%), and lower protein levels (mean 0.39 vs 1.12 g/l) on cerebrospinal fluid analysis. In the neuroborreliosis group, the duration of symptoms was shorter and the cerebrospinal protein level was lower in cases of LM associated with PFP compared to isolated LM (mean 1.3 vs 15 days; mean 0.55 g/l vs 1.12 g/l). Conclusions. Our results suggest that some clinical data and laboratory findings may help the physician to diagnose aseptic or Lyme meningitis before completion of serologic testing. LM should be suspected in cases of meningitis with very low CSF neutrophilic counts and high protein levels associated with prolonged duration of symptoms, low grade fever, and absence of pronounced signs of meningitis. PMID- 12655418 TI - Frequency of mtDNA A1555G and A7445G mutations among children with prelingual deafness in Turkey. AB - Considerable differences on the frequencies of the mitochondrial 12S rRNA A1555G and tRNA(Ser(UCN)) A7445G mutations have been reported in different populations. Our screening of 168 patients coming from independent Turkish families with prelingual sensorineural non-syndromic deafness revealed three deaf children with A1555G (1.8%) but no examples of A7445G. One proband with the mitochondrial A1555G mutation has also evidence for right parietal infarct on a brain imaging study, for which common thrombotic mutations were found to be negative. CONCLUSION: This study shows that the mitochondrial A1555G mutation is among the significant causes of prelingual non-syndromic deafness in the Turkish population. PMID- 12655419 TI - Preschool development of very low birth weight children born 1994-1995. AB - This study reports on the developmental outcome of a sample of pre-term children with a birth weight < or =1500 g born in 1994 and 1995 studied at the age of 5 to 6 years. The sample included 60 out of 81 surviving pre-term children (74.1%) of a German neonatal intensive care unit which was matched to a control group of 60 kindergarten children of the same region. The results show significant differences between term and pre-term children with regard to their intellectual development, language comprehension, attentiveness, and hyperactivity. Moreover, the pre-term children required more early intervention to compensate for developmental problems. The developmental deficits were intensified by the presence of biological risk factors like increasingly low birth weight, bronchopulmonary dysplasia and intraventricular haemorrhage. CONCLUSION: differences between term and pre-term children still exist, although the early 1990s witnessed fundamental changes in the treatment of pre-term children. The persistence of these differences is explained mainly by the increase in the survival rate of children with high biological risk factors. PMID- 12655420 TI - Tracheal bronchus associated with VACTERL. AB - Tracheal bronchus (TB) associated with VACTERL has not been reported previously. A 5-month-old girl with VACTERL association was ventilator-dependent following surgical closure of a patent ductus arteriosus (PDA). Chest radiographs showed persistent hyperinflation of the right upper lobe. Bronchoscopy showed a laterally displaced right main bronchus with stenosis and malacia of the left main bronchus. Dynamic bronchography revealed a TB associated with tracheal stenosis and malacia below the abnormal bronchus. TB may represent an additional tracheal (T) anomaly in VACTERL association. Dynamic bronchography is a useful diagnostic tool in tracheobronchial anomalies. CONCLUSION: TB may represent an "associated" tracheal (T) anomaly in children with VACTERL association. Bronchoscopy may fail to diagnose an ectopic bronchus. Dynamic bronchography is a useful diagnostic tool in tracheobronchial anomalies. PMID- 12655422 TI - Refractory iron-deficiency anaemia due to silent Helicobacter pylori gastritis in children. AB - We describe the cases of three children with chronic active Helicobacter pylori gastritis and iron-deficiency anaemia without evidence of oesophagogastrointestinal bleeding. In all cases, long-standing iron supplementation became effective only after eradication of Helicobacter pylori. CONCLUSION: Iron-deficiency anaemia may be due to clinically inapparent H. pylori gastritis. PMID- 12655421 TI - The effect of hydrotherapy on the incidence of common cold episodes in children: a randomised clinical trial. AB - Few interventions have proved to be effective in the prevention and treatment of common colds. Anecdotal reports suggest the possible beneficial effect of hydrotherapy (stimulation of the body surface with warm and cold water). This study's objective was to evaluate the clinical effect of hydrotherapy on common colds in children. Children aged 3-7 years with six or more common cold episodes during the preceding 12 months were randomised to receive either daily inhalation of normal saline in the control group or daily inhalation plus daily hydrotherapy in the experimental group for 12 months. The main outcome measurements were incidence, duration and severity of common cold episodes as reported by the children's parents in a daily symptom diary. Groups did not differ at baseline with regard to age, gender, or number of cold episodes in the year before the study. Diaries were available from 81 patients in the control group and 65 patients in the experimental group. In the study period, there were no significant differences in the incidence of colds (control vs. experimental group, mean +/- SD, 4.8+/-3.5 vs. 4.1+/-3.3 episodes) or the average duration of episodes (7.7+/-3.5 vs. 7.6+/-3.8 days). CONCLUSION: This study does not demonstrate any beneficial effect of hydrotherapy on preschool children with frequent common colds. PMID- 12655423 TI - Pulmonary re-occurrence of post-transplant lymphoproliferative disease with hypogammaglobulinaemia. AB - We report the case of a 12-year-old boy, who developed Epstein-Barr virus (EBV) associated post-transplant lymphoproliferative disease (PTLD) 7 years after renal transplantation. He responded well to the reduced immunosuppressive therapy and treatment with ganciclovir. Two years later he developed severe pneumonia and hypogammaglobulinaemia related to EBV infection exacerbation. An X-ray film revealed persistent pneumonia in the right lung. Lung biopsy showed a large, diffuse EBV-associated B-cell lymphoma. This constellation suggested re occurrence of the primary PTLD. CONCLUSION: We present a case of recurring Epstein-Barr virus-associated post-transplant lymphoproliferative disease with a remarkably late onset in addition to hypogammaglobulinaemia. PMID- 12655424 TI - Causes and consequences of pedestrian injuries in children. AB - Pedestrian-car collisions are in motorised countries one of the leading causes of unintentional-injury-related death and morbidity among children. The objective of this study was to assess correlations between the mode and the severity or post traumatic sequels of the injury. A 12-year (1989-2001) retrospective study of children aged 0-16 years injured in pedestrian-motor-vehicle collisions was undertaken. An anonymous survey with questionnaires completed by families was conducted. In this time period, 501 children (average age: 7.9 years) were injured. The lack of visibility of the vehicle (45%) and/or the pedestrian (39%) was the leading cause of injury. The mean injury severity score was 6.0 (1-75) and the mortality rate was 0.8% ( n=4, injury severity scores 47, 75, 75, 75). In this study, 217 questionnaires were analysed. When the children were injured, 37% of them were alone, 32.3% were in the presence of accompanying adults, and 24.0% were in the presence of other children. The latter group suffered the highest injury severity score ( P=0.016). Positive correlations were noted between injury severity score and impact speed ( r=0.31, P<0.001) and the distances that the children were thrown off or transported on the vehicle front ( r=0.29, P<0.001). Physical sequels were reported by 27% of the families, and 23% noted psychological disturbances. A positive correlation was noted between impact speed and the occurrence of physical sequels ( P<0.001). The number of deformed parts per vehicle was correlated to the occurrence of behavioural disturbances ( P<0.004). CONCLUSION: The incidence of post-traumatic sequels seems to be associated with higher speed at impact, lack of braking attempts, and higher numbers of vehicle parts deformed. PMID- 12655425 TI - Epidemiological and clinical aspects of congenital heart disease in children in Tuzla Canton, Bosnia-Herzegovina. AB - Congenital heart disease (CHD) is among the most frequent of all congenital anomalies. The purpose of this study was to present the results of an initial registration of children with CHD from January 1994 to December 1999 in Tuzla Canton, Bosnia-Herzegovina. The population studied consisted of all 39,699 live born children in this area. Diagnosis of CHD was made by clinical findings, electrocardiography, chest X-ray, echocardiography, catheterisation or autopsy. In the 6-year period, 243 children were found to have CHD, i.e. a prevalence of 6.12 per 1000 live-born. Critical CHD was present in 58 of them, or 1.46 children per 1000 live-born. The average age at diagnosis was 1.47 years. There were 132 boys (54.3%) and 111 girls. The most frequent anomaly was ventricular septum defect with a prevalence of 2.49 per 1000 live-born, representing 40.7 % of the total anomalies. Of the total group, 46 (18.9%) had extracardiac anomalies related to syndromes. Cardiac surgery was indicated in 98 patients (40.3%) but could only be carried out in 42 (17.3%). A total of 63 (25.9%) patients died, 54 of whom within the 1st month of life. CONCLUSION: congenital heart disease is a very significant health problem in Tuzla Canton. It requires urgent measures in terms of organisation of early diagnosis and proper management. PMID- 12655426 TI - Editorial comment. PMID- 12655427 TI - Sodium valproate and measles virus replication. PMID- 12655428 TI - A premature newborn with vesiculobullous skin lesions. AB - Although the rate of congenital syphilis is declining in Western Europe and in the USA, a significant increase is observed in the developing countries. In this contribution, a newborn with cutaneous manifestations of congenital syphilis is presented. CONCLUSION: Pediatricians should be attentive when confronted with vesiculobullous skin lesions in a newborn, and congenital syphilis should be considered in the differential diagnosis. PMID- 12655429 TI - Psychotic symptoms in severe MTHFR deficiency and their successful treatment with betaine. PMID- 12655430 TI - An interim report of the pilot study of screening for congenital hypothyroidism in Tehran and Damavand using cord blood spot samples. PMID- 12655432 TI - Occurrence of coeliac disease in children with Down's syndrome in north Moravia, Czech Republic. PMID- 12655431 TI - Small thymus at birth and neonatal outcome in very-low-birth-weight infants. PMID- 12655433 TI - IgG2, IgG4 and IgA deficiency possibly associated with carbamazepine treatment. PMID- 12655434 TI - Nemaline myopathy and early respiratory failure. PMID- 12655435 TI - Acute tumor lysis syndrome in solid tumors--a case report and review of the literature. AB - PURPOSE: Tumor lysis syndrome (TLS) is a potential complication in cancer therapy. It may occur in highly sensitive tumors, especially in childhood cancers and acute leukemias, whereas it is rare in the treatment of adult solid tumors. TLS is characterized by hyperuricemia, hyperkalemia, hyperphosphatemia and hypocalcemia following massive lysis of malignant cells. Complications include acute renal failure and metabolic acidosis. We report the first case of TLS during chemotherapy in a patient with metastatic medulloblastoma, together with a review of the literature regarding the occurrence of TLS in patients with solid tumors. METHODS: Data regarding clinical and biochemical parameters were extracted from the actual patients' files. Reports of TLS in the English language literature up to 2002 were identified by searching Medline. RESULTS: A 23-year old male with metastatic medulloblastoma received chemotherapy with cisplatin and etoposide due to massive extracerebral manifestations including metastases to the liver, mediastinal lymph nodes and bone marrow metastases. The patient developed classical signs of TLS on the second day of chemotherapy, including acute renal failure. A 17-fold increase in plasma LDH up to 87608 U/l was observed together with a 4-fold increase in plasma creatinine. The patient was treated with aggressive hydration, allopurinol and repeated hemodialysis. During the following days the patient improved and the biochemical markers all returned to normal. REVIEW. Reviewing the literature, a total of 45 patients with solid tumors who developed TLS have been reported. Most of the patients presented with metastatic, therapy-sensitive disease. Although preventable in practically 100% of patients, TLS is a potentially fatal complication, and in this material the mortality rate was one in three. Risk factors included increased LDH, hyperuricemia and pretreatment azotemia. CONCLUSIONS: TLS is only rarely associated with treatment of solid tumors. Precautions should be taken to avoid this potentially fatal complication in (chemo)therapy of solid tumors, especially in therapy-sensitive tumors presenting with bulky, metastatic disease and preexisting risk factors, including azotemia, elevated LDH and hyperuricemia. Prophylactic treatment to avoid TLS includes allopurinol, hydration prior to treatment and alkalization of the urine. Urate oxidase (rasburicase) is now beginning to replace allopurinol as a more effective way of reducing hyperuricemia and thereby the risk of TLS. PMID- 12655436 TI - Toxicity of a quinocarmycin analog, DX-52-1, in rats and dogs in relation to clinical outcome. AB - PURPOSE: Quinocarmycin analog DX-52-1 is a cyanated derivative of quinocarmycin, a compound isolated from cultures of Streptomyces melanovinaceus. DX-52-1 was selected for preclinical development because it showed efficacy against melanoma cell lines in the NCI human tumor cell screen and melanoma xenografts in mice. This report describes studies in rats and dogs to determine the maximum tolerated dose (MTD) and identify dose-limiting toxicities (DLT) in each species in different regimens to establish a safe starting dose and potential target organs of DX-52-1 for phase I clinical trials. METHODS: DX-52-1 was administered to Fischer 344 rats using repeated intravenous (i.v.) slow bolus injections following q3hx3 and q3hx3,q7dx3 regimens, and to beagle dogs using a single injection, 6-h continuous i.v. infusion (c.i.v.) and weekly 6-h c.i.v. for 3 weeks. Endpoints evaluated included clinical observations, body weights, hematology, serum clinical chemistry, and microscopic pathology of tissues. RESULTS: The MTD of DX-52-1 was a total dose of 18 mg/m(2) body surface area for q3hx3 administration in rats and 30 mg/m(2) for a single c.i.v. administration in dogs. The total dose MTD for rats on a weekly (q3hx3,q7dx3) regimen was 54 mg/m(2), and for dogs on the weekly x3 (6-h c.i.v.) infusion was 60 mg/m(2). In rats, significant elevations in blood urea nitrogen and creatinine were observed together with acute renal tubular necrosis histologically. Modest increases in liver enzymes were also observed, as were decreases in reticulocytes that were unaccompanied by histologic changes in liver and bone marrow. In dogs, adverse signs included vomiting/retching, diarrhea, and transient hypothermia; also red blood cells, hemoglobin, hematocrit, and lymphocytes were decreased. Histologic evaluation of tissues from dogs revealed necrosis and cellular depletion of the bone marrow, and extensive damage to the entire gastrointestinal tract, including marked cellular necrosis of the mucosa and lymphoid necrosis of the gastrointestinal associated lymphoid tissue. Destruction of the mucosal lining of the intestinal tract was likely responsible for dehydration, toxemia, septicemia, and shock seen in moribund dogs. CONCLUSIONS: The MTD values were comparable between rats and dogs given roughly similar dose regimens (single dose or weekly) and both species tolerated a higher total dose with weekly administration. However, the principal target organ responsible for DLT in rats was the kidney, whereas in dogs, the most severe effects were on the gastrointestinal tract and bone marrow. Both renal and gastrointestinal toxicities were reported in patients after 6-h c.i.v. infusions in a limited phase I clinical trial, indicating that neither animal model alone was predictive of DX-52-1-induced toxicity in humans, and that both species were required to define human toxicity. PMID- 12655437 TI - Ritterazine B, a new cytotoxic natural compound, induces apoptosis in cancer cells. AB - PURPOSE: Ritterazine B, one of the ritterazine analogues extracted from Ritterella tokioka, has been shown to be chemically similar to cephalostatin 1, and among the ritterazine derivatives is the most cytotoxic to P388 murine leukemia cells. The objective of this study was to determine the cytotoxicity of ritterazine B to non-small-cell lung cancer (NSCLC) cells in vitro and its effects on the cell cycle and apoptosis. METHODS: The cytotoxicity of ritterazine B against PC14 NSCLC cells was investigated using a 4-day MTT assay. Morphological changes in cells after exposure to this compound were evaluated by phase-contrast microscopy. The effects on the cell cycle of HL-60 leukemia cells and PC14 cells were elucidated by flow cytometry and an in vitro CDK/cyclin kinase assay. Induction of apoptosis in HL-60 cells was assessed using the TUNEL assay and Hoechst 33342 staining. In addition, molecules involved in apoptosis were evaluated by Western blotting. RESULTS: Ritterazine B exerted strong cytotoxic effects against PC14 cells with a mean GI(50) of 75.1 n M. Cell cycle analysis showed that ritterazine B caused accumulation of HL-60 and PC14 cells at the G2/M checkpoint. Furthermore, ritterazine B-treated HL-60 cells became multinucleated, and at a concentration of 20 n M this resulted in the onset of apoptosis. Neither cleavage of caspase target molecules nor phosphorylation of bcl-2 were observed in ritterazine B-treated HL-60 cells. CONCLUSIONS: These results indicate that ritterazine B might be a potent inducer of apoptosis acting via a novel antimitotic mechanism. PMID- 12655438 TI - Growth suppression of a tumorigenic rat liver cell line by the anticancer agent, ET-18-O-CH(3), is mediated by inhibition of cytokinesis. AB - PURPOSE: This study was undertaken to elucidate the potential mechanism of the antitumor activity of ET-18-O-CH(3), a synthetic analogue of lysophosphatidyl choline, and a known antitumor agent and specific inhibitor of phosphoinositide phospholipase C (PI-PLC). METHODS: A normal rat liver epithelial "oval" cell line (WB-F344) was neoplastically transformed by the H-ras oncogene (WB-ras2) and treated with a series of ET-18-O-CH(3) concentrates for a number of days. Cell growth, morphological "differentiation", cell cycle regulation, karyotypic changes, growth in soft agar (anchorage-independent growth) and the expression of cdk2, cdc2 and ERK genes were studied to determine the effect of ET-18-O-CH(3) on these neoplastic cells. RESULTS: ET-18-O-CH(3) at 5 and 10 microg/ml was found to cause an increase in cell size, suppress cell growth, reduce the colony-forming efficiency and inhibit the anchorage-independent growth of the WB-ras2 cells. Significantly, flow-cytometric analysis revealed that while control cells and cells treated with concentrations of ET-18-O-CH(3) below 5 microg/ml were diploid, cell populations treated with 5 and 10 microg/ml ET-18-O-CH(3) comprised 33-37% diploid cells and over 60% tetraploid cells (4n-8n cycle cells). ET-18-O CH(3) was found to induce aberrant cytokinesis as evidenced by the presence of a high frequency of enlarged cells, which were binucleated or multinucleated and mitotic cells with 4n and 8n numbers of chromosomes. ET-18-O-CH(3) was also capable of inhibiting both the expression of cdk2 and cdc2 and the activation of ERK1/2, while no effect was found on the expression of p21 ras. CONCLUSIONS: The effect of ET-18-O-CH(3) on neoplastically transformed H-ras rat liver cells has been interpreted as the result of an altered phenotype characterized by an enlarged and flattened cell morphology with ploidy changes caused by inhibition of cytokinesis. PMID- 12655439 TI - Stimulation of intestinal epithelial restitution by prostaglandin E(1) analogue. AB - BACKGROUND: 5-Fluorouracil (5-FU) causes intestinal mucosal damage and malabsorption. We have recently reported that coadministration of 17 S,20 dimethyl- trans- lower right triangle (2)-prostaglandin E(1) (OP-1206), a stable synthetic analogue of prostaglandin E(1), with 5-FU to rats protects the small intestine from 5-FU-induced damage. Enterocyte proliferation would contribute to the restitution of the wounded intestinal mucosa. Thus, we investigated the effect of OP-1206 on the proliferation of rat jejunal crypt cells (IEC-6 cells) treated with 5-FU. METHODS: Proliferation of IEC-6 cells was evaluated in terms of [(3)H]-thymidine incorporation and using the 3-(4,5-dimethyl-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) assay. Mucosal healing was assessed by measuring the speed of resealing across the denuded area of an IEC-6 cell monolayer. RESULTS: OP-1206 stimulated [(3)H]-thymidine incorporation into subconfluent IEC-6 cells pretreated with 5-FU and increased the number of IEC-6 cells. AH23848B, an EP4 prostaglandin receptor antagonist, blocked the OP-1206 stimulated [(3)H]-thymidine incorporation into IEC-6 cells. The speed of resealing across the denuded area of a wounded IEC-6 cell monolayer was found to increase following treatment with OP-1206. CONCLUSIONS: OP-1206 stimulated the proliferation of IEC-6 cells treated with 5-FU, indicating a possible mechanism for the protective effect of OP-1206 against 5-FU-induced damage to the small intestine. OP-1206 was shown to be active in intestinal mucosal healing. PMID- 12655440 TI - Comparison of the sulforhodamine B assay and the clonogenic assay for in vitro chemoradiation studies. AB - PURPOSE: Since there is a growing interest in preclinical research on interactions between radiation and cytotoxic agents, this study focused on the development of an alternative to the very laborious clonogenic assay (CA). METHODS: The colorimetric sulforhodamine B (SRB) assay was compared to the clonogenic assay for radiosensitivity testing in two lung cancer cell lines (A549, H292), one colon cancer cell line (HT-29) and one breast cancer cell line (MCF-7). In addition, the combination of the radiosensitizing agent gemcitabine and radiation was investigated with both assays. RESULTS: The dose-response curves obtained with the SRB assay and the CA were very similar up to 6 Gy. The radiosensitivity parameters (SF(2), alpha, beta, MID and ID(50)) obtained from the SRB assay and the CA were not significantly different between H292, A549 and MCF-7 cells. The radiation dose-response curves for A549 and H292 cells pretreated with 4 n M gemcitabine for 24 h clearly showed a radiosensitizing effect with both assays. The dose-enhancement factors obtained with the SRB assay and the CA were 1.80 and 1.76, respectively, for A549 cells, and 1.52 and 1.41 for H292 cells. CONCLUSIONS: The SRB assay was shown to be as useful as the more traditional CA for research on chemotherapy/radiotherapy interactions in cell lines with moderate radiosensitivity. This assay will be used for more extensive in vitro research on radiosensitizing compounds in these cell lines. PMID- 12655441 TI - Phase I and pharmacokinetic study of anhydrovinblastine every 3 weeks in patients with refractory solid tumors. AB - PURPOSE: Anhydrovinblastine (AVLB) is a novel semisynthetic vinca alkaloid. We conducted a phase I trial to determine the maximum tolerated dose (MTD), dose limiting toxicities (DLT) and pharmacokinetics of AVLB given as a 1-h intravenous infusion once every 3 weeks in patients with advanced refractory solid tumors. PATIENTS AND METHODS: Entered into the study were 24 patients with normal bone marrow, hepatic and renal function, and of these 21 were evaluable. There were 12 males and 12 females with a median age of 60 years (range 27-75 years). Diagnoses were non-small-cell lung cancer (NSCLC) (11), colorectal cancer (5), soft tissue sarcoma (4), and miscellaneous (4). Patients had had a median of three prior chemotherapy regimens (range one to six). A total of 51 courses were administered at doses of 2.5, 5, 10, 16.5, 21, 25 and 30 mg/m(2) in one, three, one, three, six, six and one patient respectively. RESULTS: Grade 2 infusional hypertension, anemia, and dizziness were noted at 16.5 mg/m(2). At 25 mg/m(2), two of six evaluable patients had DLT. DLT was grade 4 constipation, neutropenia and grade 3 nausea/vomiting. At 21 mg/m(2) one of six evaluable patients had DLT (grade 3 nausea/vomiting). This dose was the MTD. Stable disease was noted in one patient with metastatic sarcoma to the lungs and in three patients with metastatic NSCLC. The pharmacokinetics of AVLB were linear, and well characterized by a two compartment model, with a mean clearance of 26.4 l/h per m(2) and median terminal half-life of 18 h. CONCLUSIONS: The recommended phase II dose is 21 mg/m(2). A phase II study in NSCLC is being initiated. PMID- 12655443 TI - Intermittent FLDP: 24-h infusion of 5-FU on days 1, 3 and 5 combined with low dose cisplatin on days 1-5 for gastric cancer, and its pharmacologic and kinetic rationale. AB - PURPOSE: To improve the therapeutic efficacy and minimize the toxicity of 5 fluorouracil (5-FU), intermittent therapy consisting of alternate 24-h intravenous infusion and based on differences in generation time (T(G)) between normal cells and tumor cells was investigated. METHODS: Two human gastric cancer cell lines MKN-7 and MKN-74 with T(G) of 35 h and 17 h, respectively, were used in an in vitro cytotoxic assay. The drug exposure schedule consisted of a continuous 144-h exposure and alternate 24-h exposures. In a clinical trial, a total of 23 patients with advanced or recurrent gastric cancer were treated with intermittent therapy consisting of 24-h intravenous infusion with 5-FU 700 mg/m(2) per day on days 1, 3 and 5 in combination with low-dose cisplatin (CDDP) at 3.3 mg/m(2) per day on days 1 to 5. One cycle of the combined chemotherapy lasted for four consecutive weeks, followed by withdrawal over 1-2 weeks. Plasma 5-FU concentrations were measured by high-performance liquid chromatography in 15 patients and dihydropyrimidine dehydrogenase (DPD) activity in peripheral blood mononuclear cells (PBMC) was measured in 13 patients. RESULTS: The in vitro study revealed no statistically significant difference in cytotoxicity of 5-FU between the two drug exposure schedules in MKN-7 cells. In MKN-74 cells, however, a statistically significant decrease in cytotoxicity was found with the alternate 24-h exposure. In a clinical trial, plasma 5-FU concentrations showed a trapezoidal pattern. There was a significant correlation between DPD activity in PBMC and total body clearance of 5-FU. There were eight partial responders (8/22, 36%). Toxicities were very mild in severity, with no grade 3 or 4 toxicity. In particular, diarrhea and stomatitis were infrequent (one patient), and none of the patients developed thrombocytopenia. CONCLUSIONS: Toxicities which may be observed in rapidly growing cells such as bone marrow cells and gastrointestinal epithelial cells following continuous intravenous infusion of 5-FU seemed to be reduced by intermittent therapy of 5-FU consisting of alternate 24-h intravenous infusions. PMID- 12655442 TI - Phase I trial of continuous infusion 5-aza-2'-deoxycytidine. AB - PURPOSE: To identify a dose of the demethylating agent 5-aza-2'-deoxycytidine (DAC) with acceptable side effects, and to study its effect on the methylation patterns of relevant genes in tumor biopsies before and after treatment with a novel methylation assay using real-time PCR. METHODS: A group of 19 patients with metastatic solid tumors were treated with DAC by continuous intravenous infusion over 72 h, days 1-3 of a 28-day cycle. Tumor biopsies were taken before and 7 days after starting DAC. RESULTS: The dose levels studied were 20, 30 and 40 mg/m(2). Grade 4 neutropenia was found in two of five patients at 40 mg/m(2) and one of six patients at 30 mg/m(2). No objective responses were seen in this study. Steady-state DAC levels of 0.1 to 0.2 microM were achieved in the 30 and 40 mg/m(2) cohorts. Changes in methylation were observed, but no single gene consistently demonstrated evidence of demethylation. CONCLUSIONS: DAC was tolerated at a dose of 30 mg/m(2) per day for a 72-h intravenous infusion. Changes in gene methylation were observed. PMID- 12655444 TI - Antitumor activity of Virulizin, a novel biological response modifier (BRM) in a panel of human pancreatic cancer and melanoma xenografts. AB - PURPOSE: To define the anticancer efficacy of Virulizin in vivo as a single agent or in combination with conventional drugs in human pancreatic tumor and melanoma xenografts. METHODS: The therapeutic effect of Virulizin was evaluated in a series of human tumor xenografts in athymic nude mice. RESULTS: Virulizin had a high level of antitumor activity against all the pancreatic tumors (BxPC-3, SU 86.86. and Mia-PaCa-2) and melanomas (C8161 and A2058), as indicated by suppression of tumor growth with an optimal T/C value of 99% ee. ( S)-ECHB was synthesized to 48.7 g/l with an optical purity of 99.8% ee, using recombinant E. coli cells coexpressing AR from R. eutropha and glucose dehydrogenase from B. subtilis for the regeneration of NADPH. PMID- 12655456 TI - Molecular cloning, sequencing and expression of the gene encoding a novel chitinase A from a marine bacterium, Pseudomonas sp PE2, and its domain structure. AB - The pchA gene encoding chitinase A (PchA) from a Pythium porphyrae cell-wall degrading marine bacterium, Pseudomonas sp. PE2, was cloned and characterized. The deduced PchA was a modular enzyme composed of an N-terminal signal peptide, a glycoside hydrolase family 18 catalytic domain that was responsible for the chitinase activity, the chitin-binding domains (ChBDs), and the carbohydrate binding modules (CBM). The amino acid sequence of ChBD(PchA) was highly conserved in the CBM family 12 that also accommodates ChBDs without an AKWWTQG motif, a domain commonly found in bacterial chitinase and Streptomyces griseus protease C. Interestingly, CBM(PchA) showed significant sequence homology to the C-terminal region of endoglucanase B from Cellvibrio mixtus, which is a member of CBM family 6. This is the first report of a chitinase possessing a domain with high similarity to CBM family 6. Deletion analysis indicated clearly that ChBD(PchA) might play an important role in the binding of native chitin and chitosan, but not processed chitin. CBM(PchA) also appeared to play such a role in the binding of xylan and Avicel. These results suggest that the C-terminal region of PchA might be a key component in the binding of chitin in the cell walls of P. porphyrae or other structural components of marine organisms. PMID- 12655457 TI - Transformation and mineralization of 2,4,6-trinitrotoluene by the white rot fungus Irpex lacteus. AB - Unlike other 2,4,6-trinitrotoluene (TNT)-degrading white rot fungi, including Phanerochaete chrysoporium, initial metabolism of TNT by Irpex lacteus was found to occur through two different transformation pathways. Metabolites of the nitro group reduction pathway were confirmed with the standard compounds, and the formation of hydride-Meisenheimer complex of TNT (H(-)-TNT) formed in the denitration pathway was identified with LC/MS and by LC/photodiode array (PDA) detection. The molecular weight of the H(-)-TNT complex was identified as 228 m/z, and the UV-visible absorption spectrum, recorded with a PDA detector, proved the identity of this metabolite (RT, 18.7 min; lambda(max) 254, 474, 557 nm) by comparison with the authentic synthetic H(-)-TNT (RT 18.7 min; lambda(max) 261, 474, 563 nm). Mineralization of [U-(14)C]TNT by I. lacteus was also measured in static and shaken cultures. The mineralization rate of TNT in the static culture was higher than that in the shaken culture, and addition of Tween 80 (final concentration 1%) enhanced the mineralization of TNT in the static culture, resulting in 30.57% of CO(2) evolution from the radiolabeled TNT added. The high TNT mineralization capability of I. lacteus seemed to be the result of simultaneous utilization of the denitration pathway, which is more favorable for the ring cleavage and mineralization of TNT, together with the nitro group reduction pathway. PMID- 12655458 TI - Brettanomyces bruxellensis: effect of oxygen on growth and acetic acid production. AB - The influence of the oxygen supply on the growth, acetic acid and ethanol production by Brettanomyces bruxellensis in a glucose medium was investigated with different air flow rates in the range 0-300 l h(-1 ) x (0-0.5 vvm). This study shows that growth of this yeast is stimulated by moderate aeration. The optimal oxygen supply for cellular synthesis was an oxygen transfer rate (OTR) of 43 mg O(2) l(-1) x h(-1). In this case, there was an air flow rate of 60 l h(-1) (0.1 vvm). Above this value, the maximum biomass concentration decreased. Ethanol and acetic acid production was also dependent on the level of aeration: the higher the oxygen supply, the greater the acetic acid production and the lower the ethanol production. At the highest aeration rates, we observed a strong inhibition of the ethanol yield. Over 180 l h(-1) x (0.3 vvm, OTR =105 mg O(2) l( 1) x h(-1)), glucose consumption was inhibited and a high concentration of acetic acid (6.0 g x l(-1)) was produced. The ratio of "ethanol + acetic acid" produced per mole of consumed glucose using carbon balance calculations was analyzed. It was shown that this ratio remained constant in all cases. This makes it possible to establish a stoichiometric equation between oxygen supply and metabolite production. PMID- 12655459 TI - Global metabolic regulation analysis for Escherichia coli K12 based on protein expression by 2-dimensional electrophoresis and enzyme activity measurement. AB - Regulation of the main metabolic pathways of Escherichia coli K12 was investigated based on 2-dimensional electrophoresis (2DE) and the measurement of enzyme activities. The cells were grown aerobically in different carbon sources, such as glucose, acetate, gluconate or glycerol. Microaerobic cultivation was also conducted with glucose as a carbon source. Fifty-two proteins could be identified based on 2DE, and 26 enzyme activities from the main metabolic pathways-including glycolysis, pentose phosphate pathway, TCA cycle, Entner Doudoroff pathway and fermentative pathway-were assayed. These enzyme activities, together with global and quantitative protein expression, gave us a clear picture of metabolic regulation. The results show that, compared with the control experiment with glucose as a carbon source under aerobic conditions, glycolytic enzymes were slightly up-regulated (<2-fold), TCA cycle enzymes were significantly down-regulated (2- to 10-fold), and fermentative enzymes such as pfl and adhE were highly up-regulated (>10-fold) under microaerobic conditions in glucose medium. When acetate was used as a carbon source, pfkA, pykF, ppc and zwf were down-regulated, while fbp, pckA, ppsA and mez were significantly up regulated. Glyoxylate enzymes such as aceA and aceB were strongly up-regulated (>10-fold) and TCA-cycle-related enzymes were also up-regulated to some extent. With gluconate as a carbon source, edd, eda, fbp and TCA cycle enzymes were up regulated. With glycerol as a carbon source, fbp and TCA cycle enzymes were up regulated, while ackA was significantly down-regulated. Protein abundance obtained by 2DE correlated well with enzyme activity, with a few exceptions (e.g., isocitrate dehydrogenase), during aerobic growth on acetate. PMID- 12655460 TI - Neurochemical correlates of relapse to d-amphetamine self-administration by rats induced by stimulation of the ventral subiculum. AB - RATIONALE AND OBJECTIVES: Previous studies show that electrical stimulation of the ventral subiculum (vSub) can reinstate drug-seeking behavior in rats following extinction. This study examined whether vSub stimulation could also evoke reinitiation of d-amphetamine (d-AMPH) self-administration during voluntary abstinence following a prolonged bout of drug intake. Dynamic changes in extracellular levels of dopamine (DA) and metabolites in the nucleus accumbens (NAc) during 48-h unlimited access to d-AMPH by rats were monitored. Neurochemical correlates of relapse to d-AMPH administration induced by vSub stimulation or by experimenter administered d-AMPH infusions were also examined during voluntary abstinence in separate experiments. METHODS AND RESULTS: In vivo microdialysis using high-pressure liquid chromatography with electrochemical detector (HPLC-EC) was used to monitor changes in DA and metabolite efflux in the NAc during a continuous access-abstinence-relapse cycle of d-AMPH self administration in a 48-h test. The initial pattern of drug intake was associated with significant increases in DA efflux in the NAc. During the abstinence phase, DA efflux was near pre-session baseline values. Electrical stimulation at the vSub after 2 h of abstinence immediately induced a significant increase in DA efflux and reinstatement of drug self-administration behavior. Evoked DA release and responses on drug-paired lever induced by vSub stimulation were significantly greater than those induced by experimenter-administered d-AMPH. CONCLUSIONS: Relapse to drug-taking behavior can be triggered by activation of the subicular glutamatergic pathway to the NAc. This study also confirmed that during abstinence the neurochemical response of the mesolimbic DA system to d-AMPH is attenuated and this can be reversed by vSub stimulation. PMID- 12655461 TI - Effects of 8-OH-DPAT and WAY-100635 on performance on a time-constrained progressive-ratio schedule. AB - RATIONALE: Performance on progressive-ratio schedules has been proposed as a means of assessing the effects of drugs on motivation. We have adopted a mathematical model proposed by Killeen to analyse the effects of drugs acting at 5-HT(1A) receptors on progressive-ratio performance. According to this model, the relationship between response rate and ratio size is described by a bitonic (inverted-U) function. One parameter of the function, a, expresses the motivational or "activating" effect of the reinforcer (duration of activation of responding produced by the reinforcer), whereas another parameter, delta, expresses the minimum time needed to execute a response and is regarded as an index of "motor capacity". OBJECTIVE: To examine the effect of the selective 5 HT(1A) receptor agonist 8-OH-DPAT [8-hydroxy-2-(di- n-propylamino)tetralin] and the antagonist WAY-100635 [ N-[2-(4-[2-methoxyphenyl]-1-piperazinyl)ethyl]- N-2 pyridinylcyclo-hexanecarboxamide] on progressive-ratio schedule performance. METHODS: Sixteen rats responded for a food-pellet reinforcer on a time constrained progressive-ratio schedule (55-min sessions). In phase 1, they received single doses (s.c.) of 8-OH-DPAT (25, 50, 100, 200 microg kg(-1), four treatments at each dose) or the vehicle (0.9% saline solution). In phase 2, they received WAY-100635 (30, 100, 300 microg kg(-1)) according to the same regimen. In phase 3, they received 8-OH-DPAT (100 microg kg(-1)) alone or in combination with WAY-100635 (30 microg kg(-1)). 8-OH-DPAT dose dependently increased the value of a, significant increases being seen with the 50, 100 and 200 microg kg( 1) doses. The highest dose also increased delta. WAY-100635 did not significantly alter either a or delta. WAY-100635 significantly attenuated the effect of 8-OH DPAT on both a and delta. CONCLUSIONS: The results suggest that 8-OH-DPAT enhanced the activating effect of the reinforcer (the highest dose may also have induced motor debilitation). The finding that the effect of 8-OH-DPAT on a was attenuated by WAY-100635 implicates 5-HT(1A) receptors in this effect. The results are consistent with previous reports that 8-OH-DPAT facilitates feeding and food-reinforced operant responding in rats and suggest that these effects may be brought about by an increase in food motivation. PMID- 12655462 TI - Brain muscarinic receptor subtypes mediating water intake and Fos following cerebroventricular administration of bethanecol in rats. AB - RATIONALE: The brain regions and receptor subtypes involved in water intake following central cholinergic stimulation have been incompletely characterized. OBJECTIVES: To examine whether drinking and brain Fos-immunoreactivity (ir) induced in rats by central administration of bethanecol is reversed by either the preferential M1 antagonist pirenzepine, the M3 antagonist 4-DAMP, or their combination. METHODS: Male Sprague-Dawley rats were surgically implanted with cerebroventricular cannulae. The muscarinic agonist, bethanecol was used as the dipsogenic agent. Either nonselective (atropine) or selective muscarinic receptor antagonists were injected together with bethanecol to determine blockade of drinking. In parallel studies, Fos-ir was assessed in discrete brain regions. RESULTS: Bethanecol-induced drinking was completely blocked by atropine or by a combination of pirenzepine and 4-DAMP; these latter antagonists alone produced sub-total inhibition of drinking. In contrast, water intake induced by angiotensin II was unaffected by combination of pirenzepine and 4-DAMP. Fos-ir was induced by bethanecol in many brain regions previously implicated in body fluid regulation, including subfornical organ and the magnocellular supraoptic and paraventricular hypothalamic nuclei. Induced Fos-ir was substantially but not completely prevented by co-injection of either pirenzepine or 4-DAMP, but their combination did not seem markedly more effective than either alone. CONCLUSIONS: Drinking induced by brain muscarinic receptor stimulation seems to proceed by a combination of M1 and M3 receptor subtypes. Drinking induced by angiotensin II occurs independently of this mechanism. Fos-ir induced in fluid-related brain regions by bethanecol either uses additional receptor type(s) or is less easily blocked than drinking behavior. PMID- 12655463 TI - Perinatal lead exposure and relapse to drug-seeking behavior in the rat: a cocaine reinstatement study. AB - RATIONALE: Intravenous self-administration of cocaine at low doses is increased by chronic low-level exposure to lead during gestation and lactation (perinatal lead exposure). Insofar as drug potency is increased by early lead exposure, it must be considered that cocaine-seeking and relapse after periods of withdrawal similarly may be enhanced by perinatal lead exposure. OBJECTIVES: Employing an animal model, the present study examined the effects of lead exposure during gestation and lactation on cocaine-induced reinstatement of drug-seeking, when animals were tested as adults. METHODS: Adult female rats were gavaged once daily with 0 or 16 mg lead for 30 days prior to breeding with non-exposed males. This exposure regimen continued until offspring were weaned at postnatal day (PND) 21. At PND 120, male offspring were trained to self-administer cocaine intravenously (IV) [0.50 mg/kg cocaine per infusion on a fixed-ratio schedule where two lever presses resulted in drug delivery (FR-2 schedule)]. After steady-state responding was established, cocaine reinstatement responding was assessed for each group within an extinction paradigm. During the initial 1 h of reinstatement testing, the previous baseline contingencies were in place, i.e. animals operated under an FR-2 schedule for an infusion of 0.50 mg/kg cocaine. During the 2 h, 3 h, and 4 h of testing saline infusions were substituted for cocaine infusions. After responding extinguished during hour 4, reinstatement of responding was tested by administering an intraperitoneal (IP) priming injection of 0.00, 5.00, 10.00, or 20.00 mg/kg cocaine. Following these injections, lever responding for saline infusions was monitored during hour 5. RESULTS: The number of saline infusions self-administered during hour 5 increased in a dose-dependent fashion for both controls (group 0-mg) and lead-exposed (group 16-mg) animals. However, lead exposed animals self-administered significantly more saline infusions than controls at the 5.00 mg/kg and 10.00 mg/kg doses. This apparent metal-related increase in sensitivity to cocaine was evident with blood lead in metal-exposed test animals returning to control levels. However, brain lead levels remained elevated in lead-exposed test animals, relative to controls. CONCLUSIONS: The results of this investigation suggest that low-level lead exposure during gestation and lactation increases sensitivity to the relapse phase of drug abuse. It is further apparent that this increased sensitivity to the reinstatement of drug-seeking behavior is long-lasting. PMID- 12655466 TI - ["Where the doctors (...) handled the women beastly (...)". The Marburg lying in hospital from the view of the women]. AB - In the statement she made in 1864, the housemaid, Elizabeth Gunkel, accused of infanticide, is reported to have said that she would have preferred to have drowned herself rather than look for a lying in hospital in which to give birth to her child. Even though they were feared by pregnant women due to the dubious- and often fatal--experiments performed there, lying in hospital continued to be the only places left for unmarried, abandoned or poverty-stricken women under financial or moral pressure to resort to. The lecture supplies a graphic illustration of the dynamics of interplay taking place in the process of medicalization within the system in which pregnant women became involved--state, legal system, church, science, traditional life-styles. As such is given a vivid picture of the daily life of women in the 19th century who became pregnant not by choice--their fears, hopes, despair and resistance to an environment in which they saw little real chance for themselves. PMID- 12655465 TI - Role of acetaldehyde in ethanol-induced conditioned taste aversion in rats. AB - RATIONALE: In spite of many recent studies on the effects of acetaldehyde, it is still unclear whether acetaldehyde mediates the reinforcing and/or aversive effects of ethanol. OBJECTIVES: The present study reexamined the role of acetaldehyde in ethanol-induced conditioned taste aversion (CTA). A first experiment compared ethanol- and acetaldehyde-induced CTA. In a second experiment, cyanamide, an aldehyde dehydrogenase inhibitor, was administered before conditioning with either ethanol or acetaldehyde to investigate the effects of acetaldehyde accumulation. METHODS: A classic CTA protocol was used to associate the taste of a saccharin solution with either ethanol or acetaldehyde injections. In experiment 1, saccharin consumption was followed by injections of either ethanol (0, 0.5, 1.0, 1.5 or 2.0 g/kg) or acetaldehyde (0, 100, 170 or 300 mg/kg). In experiment 2, the rats were pretreated with either saline or cyanamide (25 mg/kg) before conditioning with either ethanol or acetaldehyde. RESULTS: Both ethanol and acetaldehyde induced significant CTA. However, ethanol produced a very strong CTA relative to acetaldehyde that induced only a weak CTA even at toxic doses. Cyanamide pretreatments significantly potentiated ethanol- but not acetaldehyde-induced CTA. CONCLUSIONS: The present results indicate that ethanol induced CTA does not result from brain acetaldehyde effects. In contrast, it is suggested that the reinforcing effects of brain acetaldehyde might actually reduce ethanol-induced CTA. Our results also suggest that the inhibition of brain catalase activity may contribute to the potentiating effects of cyanamide on ethanol-induced CTA. PMID- 12655467 TI - [Sexuality--a taboo topic?]. AB - Looking at the media one might believe there are no sexual taboos. On the contrary: People hardly discuss their own sexual experiences and behaviour. This applies in equal measure to day-to-day clinical and medical practice, although many studies definitely underline that patients do need information and counselling. The present contribution highlights a few fundamental aspects when treading on such delicate ground and provides helpful pointers. PMID- 12655468 TI - [Sexual activity in old age-problem-oriented counselling from the psychiatric point of view]. AB - Empirical investigations often report sexual activity in old age. Society's negative stereotype of the aging person, however, results in the topic being socially taboo and prejudiced. Active sexual intercourse gets less frequent in advancing age, but masturbation and sexual fantasies still prevail. In higher age groups men are seemingly more active than women. Persons living inconstant partnerships are more active than persons living on their own. There is a significant shift from sexual intercourse to tenderness, confidence, and satisfying relations. There are numerous physical, psychological, and social factors influencing sex life in old age. Important physiological alterations and their consequences are to be observed in advancing age. Multiple diseases, drugs, and objective and subjective (e.g. anxiety disorders) after-effects of operations often result in reducing or totally giving up sexual activities. If a doctor has hints pointing to sex problems these ought to be talked about without any prejudice and with an intention of looking for possible solutions. Frequently these problems are indicative of psychological or functional disorders which may respond to a treatment by competent counselor or psychotherapist. PMID- 12655469 TI - CD8+CD45RA+CD27-CD28-T-cell subset in PBL of cervical cancer patients representing CD8+T-cells being able to recognize cervical cancer associated antigens provided by HPV 16 E7. AB - OBJECTIVE: In response to antigenic stimulation, naive MHC-class I restricted and antigen-specific CD8+CD45RA+CD28+T-cells undergo clonal expansion and differentiate into CD8+CD45RO+ memory T-cells. Upon re- encounter with the nominal antigen, CD45RO+ T-cells are able to convert to CD8+CD45RA+CD28-T-cells displaying potent immune effector functions, including TNF-alpha production. This T-cell subpopulation constitutes a minor population in healthy individuals. In the present study we are currently evaluating whether this particular T-cell subset in PBL represents CD8+T-cells which may be able to recognize cervical cancer associated antigens provided by HPV 16 E7. MATERIAL AND METHODS: Flow cytometric cell sorted CD8+CD45RA+CD28- and CD8+CD45RA+CD28-T-cells were obtained from patients with cervical cancer and tested for recognition of HLA-A2 restricted peptides derived from the human papillomavirus (HPV)16-E7 gene product using ELISA. HPV DNA in tumor tissue was detected by PCR. RESULTS: We show that the effector CD8+CD45RA+CD28-T-cell subset is expanded in peripheral blood lymphocytes (PBL) from patients with cervical cancer, but also in PBL from patients with an acute mycobacterial infection. CD8+T-cells from 3/6 cancer patients showed a peptide-specific immune response which could be segregated in peptide epitopes which elicited either a strong TNF-alpha production, or GM-CSF and IL-2 secretion. Peptide-reactivity could exclusively be detected in the ex vivo freshly isolated CD8+CD45RA+CD28-T-cell population. A similar situation was found to be true for HLA-A2 presented peptide epitopes derived from M. tuberculosis-associated antigens presented to T-cells obtained from patients with tuberculosis. CONCLUSIONS: The sorting of CD8+CD45RA+CD28-T-cells enables to determine the fine specificity of CD8+ effector T-cells without the need for in vitro manipulation and aids to define the most appropriate target epitopes for novel vaccine designs. PMID- 12655470 TI - [Efficiency and side effects of antiretroviral treatment of HIV infected pregnant women]. AB - OBJECTIVE: Antiretroviral treatment (ART) of pregnant women and newborn reduces the vertical HIV transmission below 2 % but long term effects are still not clear. METHOD: We analyzed the data of 59 children born to HIV infected women. RESULTS: The median observation time was 404 (14-740) days. There was no vertical HIV transmission (95 %-confidence interval [CI] 0-5.1 %). Among 6 children without prenatal ART one child with congenital diaphragmatic hernia was born. In 53 infants with prenatal ART we observed 19 congenital malformations (35.8 %, 95 %-CI 23.2-50.6 %). Three were severe malformations (5.7 %, 95 %-CI 1.2-15.5 %): one supraorbital hemangioma (after zidovudine), one athetoid cerebral palsy without evidence of mitochondrial dysfunction (after zidovudine), and one unilateral femoral hypoplasia (after short term zidovudine, lamivudine, and efavirenz in late pregnancy). Sixteen minor malformations were found (30.1 %, 95 %-CI 21.2-43.8 %): ventricular septal defect (closed spontaneously after 6 months), atrial septal defect, transient and persistent pulmonary stenoses, hexadactylies, unilateral hearing impairment, dystopic kidney, subependymal bleeding cysts, and mild dilation of ventricles. CONCLUSIONS: ART prevents vertical HIV transmission. All women should be advised ART during pregnancy and delivery. The prevalence of malformations after prenatal/perinatal ART appears considerably high. Therefore, detailed information for the parents and long term follow-up of all children exposed to antiretroviral drugs are crucial. PMID- 12655471 TI - [Disturbed utero-tubal transport in hysterosalpingoscintigraphy as a predictive functional test for IVF therapy]. AB - Hysterosalpingoscintigraphy (HSSG) is a simple method to evaluate the transport function of uterus and fallopian tubes. There is a quick uptake of radionuclides into the uterus and a transport to the side bearing the dominant follicle in 70 % of the patients in the late follicular phase of the cycle. Uptake and transport of the immotile radionuclides imitate the directed sperm transport through the female genital tract at the time of ovulation. 214 of 796 infertility patients with proven patency of fallopian tubes (27 %) showed only an uptake of the particles without a transport towards the fallopian tubes (negative HSSG). In these patients no spontaneous pregnancy occurred and pregnancy rate remained low by the means of timed intercourse or insemination. Indeed, the pregnancy rate (8.4 %) was significantly lower compared with the patients who became pregnant by timed intercourse, insemination or spontaneously and had a positive HSSG before (15 %; p=0.001). However, the pregnancy rate that could only be achieved by methods of ART was significantly higher in the group of patients with negative HSSG (57 % vs. 25 %, p=0.05). Our data suggest that HSSG is a new method to evaluate the integrity of the inner genital tract's transport function, especially in patients suffering from idiopathic infertility. Impaired transport function (negative HSSG) should be considered as an indication for IVF-treatment. PMID- 12655472 TI - [Identification and characterisation of immune cells in the ejaculate of infertile and fertile men]. AB - Using the immunoperoxidase technique with monoclonal antibodies the ejaculates of 20 infertile and 15 fertile men were examined for lymphocyte subsets and activation markers to know more about the cellular mucosal immunity in the male genital tract. The results have shown that there are small differences of T helper (CD4), T-suppressor (CD8), B- (CD19) and Natural Killer cells (CD57) between both groups. The number of monocytes (CD14), activated T- and B-cells and macrophages (CD25), the LCA-positive cells (CD45) and the HLA-DR-positive cells were remarkably increased in fertile patients. It has been established that the number of NK-cells (CD57) correlated with mycoplasma infection and the B-cells (CD19) correlated with the testosterone level. It seems that the increased testosterone value improved the cellular immunity. Infertile men have a disturbed mucosal immunity and a higher rate of genital infections. PMID- 12655473 TI - Determinants of wound infections after surgery for breast cancer. AB - BACKGROUND: There are evident risks factors stemming from any the local post surgical infection, which merits analysis of morbidity derived from conservative and radical surgery after treatment of the operable breast cancer. STUDY DESIGN: 107 patients with breast cancer recruited between November 1995 and October 1997 were evaluated. From that total, 36 patients were subjected to quadrantectomy, axillary lymphadenectomy and postoperative radiation while 71 patients were subjected to modified radical mastectomy. RESULTS: 7 % percent of patients who underwent modified radical surgery succumbed to postoperative local infection vs. 17 % of patients subjected to breast conserving therapies. Local infections were associated with the older patients (63 vs. 54 years old, p=0.03) and greater obesity (p=0.04). Upon logistic regression analysis conservative surgery exhibited adjusted Odds Ratio of 6.42 (1.27-32.28), p =0.02. In postoperative infection group average hospital stay increased by 6 days (p= 0.0001). CONCLUSION: In the present study main risk factors associated with postsurgical wound infection after breast cancer treatment proved to be advanced age, greater obesity and conservative surgery. Nonetheless, more studies are necessary to confirm current findings. PMID- 12655474 TI - [Patients as "living manikins"? Gottingen University's maternity hospital ca. 1800]. AB - The maternity hospital in Gottingen, founded in 1751, is considered to be the first in the world which was part of a university. Its main purpose was to train male medical students. Secondary aims were to instruct female midwives and to provide a haven for poor pregnant and lying-in women. The hospital was open to all women, without discriminating against foreigners, or any religion or race. Almost all patients were not married, and the overwhelming majority were servants. This article makes use of printed as well as archival material, mainly from the period when Professor Friedrich Benjamin Osiander was the hospital's director, i. e. 1792-1822, in order to show how the patients were used for developing and teaching 'scientific' obstetrics and man-midwifery. PMID- 12655475 TI - [Torsion of pregnant uterus--a case report]. AB - The 29 year old woman with a caesarian section 5 years before was hospitalized in the 23 rd gestational week bleeding by a placenta praevia totalis and was intravenously tocolized. In the 26 nd gestational week a caesaran re-section was necessary because of heavy acute abdominal symptoms. The uterus presented torquated for 180 degrees to right. The male child (720 g, pH 7.22) died the next night by extreme immaturity. The mother was discharged after 8 days without complications. PMID- 12655476 TI - [Ovarian epidermoid cyst and endometrioid carcinoma: do they share their origin?]. AB - A case of a well differentiated endometrioid adenocarcinoma of the ovary in combination with an epidermoid cyst is described. A typical epidermoid cyst without skin appendages could be observed. In direct neighborhood, there was a typical endometrioid carcinoma with some foci of squamous metaplasia. There was no continuum between the squamous metaplasia of the carcinoma and the wall of the epidermoid cyst. In addition, the benign cyst and the carcinoma shared some of their immunohistochemical profile. This case highlights the still unsolved question about the origin of epidermoid cysts and adds to the hypothesis that they arise from pluripotent coelomic epithelium. PMID- 12655481 TI - The European College of Veterinary Clinical Pathology: good news for all of us. PMID- 12655482 TI - Flow cytometric detection of activated platelets in the dog. AB - BACKGROUND: Platelet activation appears to play a role in a variety of canine thrombotic disorders. At present, tests for the detection of activated platelets are not used routinely in veterinary clinical laboratories. OBJECTIVE: The purpose of this study was to develop a clinically applicable method to detect activated canine platelets. METHODS: A flow cytometric assay was developed to detect activated platelets, platelet aggregates, and platelet microparticles in the dog. Blood was collected from healthy dogs using EDTA or sodium citrate as the anticoagulant, and platelet-rich plasma was prepared. Platelets were activated by adding phorbol myristate acetate. In some experiments, platelets were fixed by incubation with 0.5% paraformaldehyde. In other experiments, platelets were stored for 4 or 24 hours at 4 degrees C before analysis. Activated platelets were detected by measuring surface expression of P-selectin and by determining the percentages of platelet aggregates and microparticles using forward-angle vs side-angle light scatter plots. Results were analyzed by using 2 way ANOVA and the Scheffe F-test. RESULTS: Platelets collected in EDTA had minimal expression of P-selectin, whereas platelets collected in sodium citrate had greater median fluorescence intensity. Fixation with 0.5% paraformaldehyde before labeling platelets with anti-P-selectin did not affect antibody binding or the percentages of platelet aggregates and microparticles. Storage of platelet rich plasma at 4 degrees C for 4 hours did not affect antibody binding or the percentages of platelet aggregates or microparticles. Activation of platelets ex vivo by addition of 10 ng/mL phorbol myristate acetate resulted in a large increase in expression of P-selectin but only slight increases in platelet aggregates and microparticles. CONCLUSION: Determination of platelet P-selectin expression and percentages of platelet aggregates and platelet microparticles may provide a clinically applicable means for detection of activated platelets in dogs. The capacity to use EDTA-anticoagulated blood samples and to fix platelets for evaluation at a later time makes the test attractive as a routine diagnostic tool. PMID- 12655483 TI - Ehlers-Danlos-like syndrome in 2 dogs: clinical, histologic, and ultrastructural findings. AB - BACKGROUND: Ehlers-Danlos syndrome comprises a group of rare inherited connective tissue diseases characterized by skin hyperextensibility, joint laxity, skin and vessel fragility, and poor wound healing. OBJECTIVE: The purpose of this report was to describe the clinical, histologic, and ultrastructural findings in 2 dogs with collagenopathies consistent with Ehlers-Danlos syndrome. METHODS: Two dogs were examined clinically; skin extensibility index was calculated. Skin biopsies obtained from the dorsum were examined by light and electron microscopy. RESULTS: Both dogs had clinical signs of skin hyperextensibility and fragility, lower skin elasticity, vessel fragility, and poor wound healing. One dog had a hip dislocation, and the other had bilateral medial patellar luxation (grade II), subcutaneous hematomas produced by minimal trauma, and generalized periodontitis. Histologic and ultrastructural examination confirmed abnormalities in the structure and arrangement of collagen fibrils. Fibroblasts were characterized by variable dilatation of the rough endoplasmic reticulum, and anomalous elastic fibers (elaunin fibers) were present in the dermis. CONCLUSION: Although the primary defects underlying collagenopathies in animals are still unknown, analysis of the ultrastructural changes in collagen fibrils and clinical findings could facilitate better characterization of these disorders in dogs. PMID- 12655484 TI - Symmetrical crusting rash in a dog. PMID- 12655485 TI - Equine immune-mediated hemolytic anemia associated with Clostridium perfringens infection. AB - BACKGROUND: Previous studies have suggested an association between equine immune mediated hemolytic anemia and clostridial infections or neck abscesses. OBJECTIVE: The purpose of this report was to describe and characterize the hematologic abnormalities in a horse with Clostridium-associated immune-mediated hemolytic anemia. We also retrospectively evaluated hematologic abnormalities in 8 horses with clostridial myositis or subcutaneous emphysema. METHODS: A 7-year old Foreign Warm-Blood gelding was evaluated for anemia and a cervical abscess. CBCs and reticulocyte counts were obtained using an Advia 120 analyzer and evaluation of Wright's-stained smears. All cases of equine Clostridium spp.myositis or subcutaneous emphysema over a 7-year period were identified in a retrospective search of the University of Minnesota Veterinary Teaching Hospital database. Clinical, hematologic, and microbiological findings were recorded. RESULTS: Clostridium perfringens genotype A was isolated from the neck wound of the gelding. The CBC was characterized by severe regenerative anemia, intravascular hemolysis, and RBC agglutination. A direct Coombs' test was positive. Moderate numbers of spheroechinocytes were observed. The total automated reticulocyte count was 4.5% (56,700/microliter), with most reticulocytes having low absorbance (mature reticulocytes). The anemia responded to penicillin and steroidal and nonsteroidal immunosuppressive drugs. Of 8 horses with myositis, all of which involved the cervical region, 5 were anemic, 1 had a positive direct Coombs' test, and 2 had increased numbers of type III echinocytes and spheroechinocytes. CONCLUSIONS: The presence of type III echinocytes or spheroechinocytes may be helpful in diagnosing immune-mediated hemolytic anemia associated with clostridial infections in horses. Automated reticulocyte counts may detect very low levels of reticulocytosis in hemolytic anemia in horses. PMID- 12655486 TI - A distal colonic mass in a dog. PMID- 12655487 TI - Metastatic balloon cell melanoma in a dog. AB - BACKGROUND: Balloon cell melanoma is a rare variant of amelanotic melanoma that is difficult to differentiate from sebaceous cell carcinoma, liposarcoma, and other clear cell neoplasms without immunohistochemistry or ultrastructural evidence of melanin or melanosomes. OBJECTIVE: The purpose of this report was to describe the clinical, cytologic, histologic, immunohistochemical, and ultrastructural findings in a dog with metastatic balloon cell melanoma. METHODS: A 6-year-old female Golden Retriever was evaluated for a white, flocculent infiltrate in the anterior chamber of the left eye and an enlarged left prescapular lymph node. Cytologic evaluation of the eye and lymph node were performed following aqueocentesis and fine-needle aspiration, respectively. The affected lymph node was examined histologically and stained for cytokeratin, vimentin, S-100, and Melan A. Following euthanasia a necropsy was performed and samples of the affected lymph node were examined by electron microscopy. RESULTS: Cytologic examination of the lymph node and aqueocentesis sample revealed round neoplastic cells that had abundant clear vacuolated cytoplasm. A tentative diagnosis of metastatic sebaceous cell carcinoma or clear cell neoplasm was made. Histologically, the affected lymph node had similar polygonal clear cells arranged in sheets and packets divided by delicate fibrovascular stroma. Immunohistochemical staining of the cells was negative for cytokeratin but positive for vimentin, weakly positive for S-100, and strongly positive for Melan A. At necropsy, metastatic lesions were identified in the diaphragm, heart, lung, kidneys, left eye, prescapular and sublumbar lymph nodes, and multiple skin sites. Ultrastructural examination of neoplastic lymph nodes revealed many membrane-bound vacuoles, myelin-like figures, and rare melanosomes. CONCLUSION: Immunohistochemical staining and ultrastructural features of the neoplastic cells supported a diagnosis of metastatic balloon cell melanoma. PMID- 12655488 TI - Mass in the laryngeal region of a dog. PMID- 12655489 TI - Serum creatinine concentrations in retired racing Greyhounds. AB - BACKGROUND: Greyhounds frequently have laboratory values that are outside reference intervals established for dogs. Our recognition of increased serum creatinine concentrations in several Greyhounds posed a problem when evaluating a Greyhound with suspected renal disease. OBJECTIVE: The purpose of this study was to compare serum creatinine concentrations between Greyhound and non-Greyhound dogs. METHODS: Thirty retired racing Greyhounds and 30 age- and gender-matched control non-Greyhound dogs were evaluated. Serum creatinine concentrations in both groups were measured using a standard biochemical method and compared statistically using a Kruskal-Wallis test. RESULTS: Creatinine concentration was significantly higher in the Greyhounds (P <.01) than in the control group. CONCLUSION: Greyhounds have a higher serum creatinine concentration than do non Greyhound dogs. This idiosyncrasy should be taken into account when evaluating healthy Greyhounds and those with suspected renal disease. PMID- 12655490 TI - Effects of MECP2 mutation type, location and X-inactivation in modulating Rett syndrome phenotype. AB - Rett syndrome (RTT) is a clinically defined disorder that describes a subset of patients with mutations in the X-linked MECP2 gene. However, there is a high degree of variability in the clinical phenotypes produced by mutations in MECP2, even amongst classical RTT patients. In a large-scale screening project, this variability has been examined by looking at the effects of mutation type, functional domain affected and X-inactivation. Mutations have been identified in 60% of RTT patients in this study (25% of whom were atypical), including 23 novel mutations and polymorphisms. More mutations were found in classical patients (63%) compared to atypical patients (44%). All of the pathogenic mutations were de novo in patients for whom parent DNA was available for screening. A composite phenotype score was developed, based on the recommendations for reporting clinical features in RTT of an international collaborative group. This score proved useful for summarising phenotypic severity, but did not correlate with mutation type, domain affected or X-inactivation, probably due to complex interactions between all three. Other correlations suggested that truncating mutations and mutations affecting the methyl-CpG-binding domain tend to lead to a more severe phenotype. Skewed X-inactivation was found in a large proportion (43%) of our patients, particularly in those with truncating mutations and mutations affecting the MBD. It is therefore likely that X-inactivation does modulate the phenotype in RTT. PMID- 12655491 TI - Self-injurious behavior in young boys with fragile X syndrome. AB - In this study, we distributed surveys to 67 families of young boys with fragile X syndrome to determine the prevalence, onset, form, function, location, and correlates of self-injurious behavior. Fifty-five surveys were completed (82%). The mean age of the boys at the time of the survey was 80 months (range = 20 144). Self-injurious behavior (SIB) was reported for 58% of the participants with a mean age of onset of 31 months. The mean number of forms of self-injury was 2 per participant. Biting was the most commonly reported form of self-injury with the fingers and back of the hand disproportionately targeted as the most prevalent self-injury body site. There was no linear increase in risk of SIB with age past 25 months. SIB was reported as most likely to occur following the presentation of difficult task demands or changes in routine. Significant group differences were found between overall ratings of problem behavior for boys with self-injury compared to those without self-injury. Groups did not differ on measures of fragile X mental retardation protein (FMRP), autism status, adaptive behavior, or age first medicated. Results are discussed in terms of future research designed to further elucidate the behavioral phenotype of fragile X syndrome. PMID- 12655492 TI - Cytogenetic and molecular characterization of a de novo 4q24qter duplication and correlation to the associated phenotype. AB - We report on a newborn with severe psychomotor retardation, minor anomalies, congenital heart defects, thumb and urogenital abnormalities. Cytogenetic analysis showed a 4q24qter duplication, never described before, as the result of a de novo t(4;14). The extension of the duplicated 4q region was defined by FISH using YAC probes. The breakpoint was localized between 106.3cM (YAC 800f2, D4S1572) and 111 cM (YAC 744e4, D4S1564). Comparing our patient with those previously reported in literature, we observed some features mature frequently reported in these patients: psychomotor retardation, retromicrognathia, low set and/or malformed ears and some more specific traits: congenital cardiac defects, hypoplastic thumb and urogenital abnormalities. PMID- 12655493 TI - Relationship of deficits of FMR1 gene specific protein with physical phenotype of fragile X males and females in pedigrees: a new perspective. AB - The effect of deficit of the FMR1-gene product (FMRP) on physical phenotype was investigated using a robust modification of the maximum likelihood estimators for pedigree data. The approach is a powerful method of examining genotype-phenotype relationships because it adjusts for intra-familial variation, and the robust modification allows violation of distributional assumptions in the data to be overcome by objectively down-weighting unusual observations. The data on 19 age- or height-adjusted physical measures including head, trunk and limb measures and height and weight from 110 extended fragile X families (including 185 fragile X males and females and 120 normal relatives) were related to the FMRP levels assessed in peripheral blood lymphocytes. A significant interaction between FMRP and age was also included in the models for some measures. The results have demonstrated a linear effect of progressively reducing levels of FMRP on the values of a majority of physical measures considered in the study. The most evident effect of FMRP deficit in sexes combined was in decreasing body height and limb length, and in increasing head height and the degree of connective tissue involvement (measured by the middle finger extension angle). Heritability estimated from the complex FMRP models showed the highest values for height and limb length, and the lowest for weight, finger extension angle and some facial measures. On the basis of the present data, a possible mechanism by which the FMRP deficit impacts physical phenotype is discussed. PMID- 12655494 TI - Clinical and epidemiological studies of amniotic deformity, adhesion, and mutilation (ADAM) sequence in a South American (ECLAMC) population. AB - Amniotic deformity, adhesion, and mutilation (ADAM) sequence is a heterogeneous condition, with a broad spectrum of anomalies, where intrinsic causes, as defect of germ plasm, vascular disruption, and disturbance of threshold boundaries of morphogens during early gastrulation, alternate with extrinsic causes as amniotic band rupture to explain the condition. This study aimed to identify which phenotypes could be considered as ADAM sequence, determine the prevalence rate, and disclose risk factors for this sequence. We identified 270 cases defined as having some skin evidence of constriction band, plus those having limb defects suggestive of ADAM sequence, among 3,020,896 live and stillborns in the years 1982 to 1998 in ECLAMC (Latin American Collaborative Study of Congenital Malformations). Half of the cases presented mutilation (reduction), and deformity (ring constriction) affecting distal parts of fingers or toes bilaterally, without associated defects. Acrania, cephalocele, typical or atypical facial clefts, eyelid coloboma, and celosomia were also observed being significantly associated with the skin lesion. One affected infant in every 11,200 births, was found with stable trends during the last 17 years. There was an excess of cases in populations living at high altitude, stillborns, and neonatal infant dead. Among ADAM cases there was an excess of mothers with a prenatal history of febrile acute illness, medication drug use, or vaginal bleeding during the first trimester of pregnancy. Higher than expected frequencies of first-born child, premature birth, low birth weight for gestational length, and non-cephalic fetal presentation were also found. The observed geographic difference in birth prevalence could be a useful indication to study specific genetic and environmental candidate factors to ADAM susceptibility. PMID- 12655495 TI - Developing culturally sensitive cancer genetics communication aids for African Americans. AB - The goal of this project was to develop educational materials to communicate genetic health information in a culturally sensitive manner. These materials were designed to communicate information about cancer risk, genetic testing options, and health management options in an African American kindred with a known BRCA1 mutation. Educational materials were pilot-tested in four African American focus groups varying in socioeconomic status and gender. The audiotaped focus groups consisted of presentation of the educational materials, followed by a feedback session led by an African American facilitator. Qualitative analysis of the focus group transcripts identified important themes and the educational materials were revised in response to the participants' suggestions. The products included a booklet and a flip chart for use in educational sessions. Focus group participants recommended a substantial reduction in technical detail, and recommended that information be personalized and made relevant to the lives of the target population. Other critical themes included the importance of building trust in the medical system and avoiding words and images that have strong negative associations in the African American community. Strategies that were successful included nontechnical images to explain genetic concepts, clip art images to energize and personalize word slides, vibrant color, identifiably African American figures, and the development of themes relevant to many African Americans. The use of these materials in an ongoing study offering BRCA1 counseling and testing to a large, rural Louisiana-based kindred will provide additional feedback about the effectiveness of the culturally tailored genetic education and counseling materials. PMID- 12655496 TI - SKY assessment of two karyotypes with 0-6 supernumerary marker/ring chromosomes and review of previously reported cases with two or more markers. AB - A 7-month-old boy with developmental delay and congenital abnormalities and a 58 year-old man with mental retardation, impaired speech, and dysmorphic features were referred for cytogenetic studies. The peripheral blood chromosome studies of Patient 1 had a de novo mosaic karyotype with 2-6 supernumerary marker chromosomes. Patient 2 had a mosaic karyotype with 1-5 supernumerary marker chromosomes and normal cells. All markers appeared to have a centromere by C banding and also by fluorescence in situ hybridization (FISH) using all centromere probe for Patient 1. The majority of the markers appeared like rings. Except for one marker in Patient 1 and 2-3 markers in Patient 2 with discernible >5 Mb euchromatin, the rest of the markers were minute and some appeared to have barely discernible euchromatin in C-banding or FISH. Spectral karyotyping (SKY) was attempted to determine the origin of the marker chromosomes. Because some markers had barely any euchromatin, their classification was not clear cut and they were identified as derived from more than one chromosome. The SKY classification of the markers in Patient 1 was 1, 3, 5, 7, 11, 15, and 22 and in Patient 2 was 1, 5, 6, or 7. Patient 2 was lost to further follow-up studies. To confirm the recurring SKY classifications in Patient 1, centromere probes for chromosomes 1, 3, 5, 7, 11, 15, and 22 were used. The markers were negative for 1, 3, and 11 but positive for 7, 15, and 22 and probably 5. Since 5 centromere probe cross hybridizes with 1 and 19, the weak signal on the marker/s in successive hybridization did not give a definitive answer. Also, the 5 paint probe was not conclusive because of the minute size of the marker. In some metaphases, two markers were derived from 5 or 22. For clinical considerations, the marker derived from 7, although variable in size, appeared to consistently have euchromatin, followed by 15, while 22 and 5 markers were mostly centromeric heterochromatin. The elastin gene probe that maps to 7q11.23, SNRPN gene that maps to 15q11.2, and TUPLE gene that maps to 22q11.2 did not give a signal on the markers. As expected for a majority of ring chromosomes, the pan telomere probe did not hybridize to any of the markers. This highly unusual karyotype was confirmed in the buccal epithelium using a mix of centromere 7 and 15 probes and the combination 14/22 probe. The ratio of additional FISH signals in the buccal mucosal cells was comparable to the ratios observed in the peripheral blood. In this study, we have attempted to consolidate the data on >/=2 marker cases to understand the analysis constraints, the range of clinical abnormalities, and the mechanisms involved. The literature was surveyed for multiple markers cases. A majority of the reported cases had two markers, either derived from the same chromosome or from two different chromosomes or two cell lines with different markers derived from the same chromosome. Cases with three or more markers were rare. The nature and extent of euchromatin content of the multiple markers appears to determine the phenotype. Frequently, multiple marker cases had small to minute markers. The clinical presentation varied from mild to severe. While two bisatellited markers may be associated with infertility, the phenotype in other cases ranged from borderline intelligence and mild dysmorphism to developmental delay, mental retardation, and congenital abnormalities. PMID- 12655497 TI - Mutation screening of FOXP2 in individuals diagnosed with autistic disorder. AB - Although it is well established that genetic factors play an important role in the etiology of autistic disorder (AD), no specific genes have as yet been implicated. Genetic epidemiological data, particularly the sharp fall in concordance rates from monozygotic to dizygotic twins, indicate that the mode of transmission of this disorder is complex and may involve several genes. The 7q31 locus has been repeatedly linked to AD, suggesting that this chromosomal region is likely to harbor a susceptibility gene for AD. Recently, variations in the FOXP2 gene were reported to be responsible for a severe speech and language disorder. Because of the chromosomal location of FOXP2 (7q31) and the putative implication of the 7q31 region both in autistic and in language disorders (a feature of AD), it has been hypothesized that FOXP2 may be implicated in the pathophysiology of AD. To test this hypothesis, we screened the FOXP2 gene coding sequence for mutations in subjects diagnosed with AD and in normal controls. We identified four silent polymorphisms that were equally distributed between patients and controls. Using an intra-family association design, we identified no transmission disequilibrium in any of the four identified alleles, suggesting that the FOXP2 gene does not play a significant role in AD. PMID- 12655498 TI - Interstitial deletion of the short arm of chromosome 1: attempt to establish a clinical phenotype (46,XX,del (1)(p22p32)). AB - A girl with a de novo interstitial deletion of the short arm of chromosome 1 (46,XX,del (1)(p22p32) is described with moderate developmental delay and minor phenotypic abnormality. These clinical manifestations are compared to previously reported patients with interstitial deletion of chromosome 1, in an attempt to identify a clinical phenotype which seems quite different from the syndrome linked to more terminal deletion of chromosome 1p, and perhaps from more proximal 1p deletion phenotype. PMID- 12655499 TI - Poland anomaly--report of an unusual family. AB - A family with classical Poland anomaly presenting in the son and a partially affected mother is reported. We hypothesize that Poland anomaly has a multifactorial basis involving vascular compromise in early development. Two possible predisposing factors are aberrant vascular formation and thrombophilia. The investigation of our family has not demonstrated an abnormality in coagulation, therefore, we surmise that an inherited structural variant of blood vessel formation is responsible. PMID- 12655500 TI - Y chromosomal polysomy: a unique case of 49,XYYYY in amniotic fluid cells. PMID- 12655501 TI - Famous people and genetic disorders: from monarchs to geniuses--a portrait of their genetic illnesses. AB - Famous people with genetic disorders have always been a subject of interest because such news feeds the curiosity the public has for celebrities. It gives further insight into their lives and provides a medical basis for any unexplained or idiosyncratic feature or behavior they exhibit. It draws admiration from society of those who excel in their specialized fields despite the impositions of their genetic illnesses and also elicits sympathy even in the most casual observer. Such news certainly catapults a rare genetic disorder into the realm of public awareness. We hereby present six famous figures: King George III, Toulouse Lautrec, Queen Victoria, Nicolo Paganini, Abraham Lincoln, and Vincent van Gogh, all of whom made a huge indelible mark in either the history of politics or that of the arts. PMID- 12655502 TI - Another explanation for familial Cornelia de Lange syndrome. PMID- 12655505 TI - Odorant receptor expression as a function of neuronal maturity in the adult rodent olfactory system. AB - Odorant receptors (ORs) are expressed in a spatially restricted manner in the mammalian olfactory epithelium (OE), and this patterning probably contributes to innervation specificity within the olfactory bulb (OB). Furthermore, glomerular targeting appears to be contingent on receptor choice. Central to the mechanism by which ORs influence axonal specificity is the timing of OR expression during the life cycle of the olfactory sensory neurons (OSNs). Data indicate that OSNs express ORs in the absence of the OB but do not address whether OR expression is an early event in OSN differentiation. Accordingly, we evaluated whether ORs are expressed in mature [olfactory marker protein (OMP(+))] and/or immature [growth associated protein of 43 kDa m.w. (GAP-43(+))] OSNs by assessing the expression of the P2 OR subtype via immunostaining for beta-gal and concurrent OMP or GAP-43 expression in P2-IRES-tauLacZ mice. Nearly 90% of P2(+) OSNs expressed OMP, whereas approximately 10% expressed GAP-43. One month after unilateral bulb ablation, the number of P2(+) OSNs decreased on the lesioned side; however, the percent of P2(+)/GAP-43(+) OSNs dramatically increased. We also determined that onset of P2 OR expression is slightly delayed when evaluated in the context of neuronal differentiation. Additionally, we defined the expression of OR(+) OSNs in the OE of rats via in situ hybridization with a panel of eight ORs followed by OMP immunostaining. All eight ORs were found in neurons situated throughout the height of the OE, including those OSNs deep to OMP staining, thus demonstrating definitively that ORs are expressed prior to the maturational state defined by OMP expression. PMID- 12655506 TI - Functional organization of vasodilator neurons in pelvic ganglia of female guinea pigs: comparison with uterine motor neurons. AB - Neurons producing vasodilation during reproductive activity constitute a large population of neurons in pelvic autonomic ganglia. We used intracellular recording, dye-filling and multiple-labeling immunohistochemistry to determine the morphology and electrophysiological properties of, and number of synaptic inputs to, vasodilator pelvic neurons in female guinea pigs. Vasodilator neurons, identified by their immunoreactivity for vasoactive intestinal peptide (VIP) and their location in paracervical ganglia, had simple dendritic arbors (1 primary dendrite) compared with nonvasodilator neurons (3 dendrites). Vasodilator neurons had more depolarized resting membrane potentials (-47 mV) than other paracervical neurons (-55 mV) and had smaller apparent cell capacitances (65 pF vs. 110 pF). Vasodilator and nonvasodilator neurons could not be distinguished on the basis of their action potential discharge characteristics or current voltage relationships. Most pelvic neurons ( approximately 70%) had tonic (slowly adapting) discharges. Fifty-five percent of vasodilator and 60% of nonvasodilator neurons showed inward rectification when hyperpolarized below -90 mV. Around 65% of neurons showed evidence of M-current. Both vasodilator and nonvasodilator neurons ( approximately 80%) expressed an A-like current. Vasodilator neurons and nonvasodilator neurons received 1-2 fast synaptic inputs following stimulation of pelvic or hypogastric nerve trunks. Most neurons received a least one strong synaptic input. These results indicate that vasodilator neurons and neighboring neurons projecting to other pelvic targets, primarily in the myometrium, express a similar range of ionic conductances and integrate few synaptic inputs. The similarities between these two populations of neurons may be related to their coactivation as part of spinal somato-pelvic reflexes. Vasodilation and uterine contraction during reproductive behavior in female guinea pigs are likely to involve input from preganglionic neurons at both lumbar and sacral spinal levels. PMID- 12655507 TI - No further loss of dorsal root ganglion cells after axotomy in p75 neurotrophin receptor knockout mice. AB - The role of the p75 neurotrophin receptor for neuronal survival after nerve crush was studied in L5 dorsal root ganglia (DRG) of knockout mice and controls with assumption-free stereological methods. Numbers of neuronal A- and B-cells were obtained using the optical fractionator and optical disector techniques. At birth, the total number of DRG neurons was 10,000 +/- 2,600 in control mice compared with 5,100 +/- 1,300 in p75 knockout mice. During postnatal development, 1,400 neuronal B-cell bodies were lost in p75 knockouts (2P < 0.05) and 1,100 in controls (NS), whereas the A-cell population remained stable. After a sciatic nerve crush, the total neuron loss in controls was 15.4% +/- 3.5% (2P < 0.05) and 22.7% +/- 5.1% (2P < 0.05) at days 14 and 42, respectively. In contrast, there was no loss in total number of neurons after crush in p75 knockout mice. Neuronal A-cell number was unchanged after the crush in p75 knockouts as well as in controls at both times. At 14 days, the population of B-cells was reduced by 24.8% +/- 3.6% in controls and by 6.1% +/- 3.5% in p75 knockouts, this difference being significant (2P < 0.001). At 42 days, the B-cell loss was 29.6% +/- 5.5% in controls and 4.2% +/- 6.4% in p75 knockouts (2P < 0.001). In conclusion, the lack of the p75 receptor results in neuronal DRG cells that are resistant to nerve injury, pointing to a role for the receptor in apoptosis. PMID- 12655508 TI - Immunohistochemical localization of citrullinated proteins in adult rat brain. AB - By using hybridoma technology, an IgM monoclonal antibody (F95) against multiple citrullinated synthetic and natural peptides was recently developed and used to stain immunohistochemically subsets of astrocytes and myelin basic protein (MBP) from selected regions of human brain (Nicholas and Whitaker [2002] Glia 37:328 336). With this antibody, the present study provides a more detailed localization of citrullinated epitopes in the central nervous system (CNS) by examining immunohistochemical staining patterns for F95 in the normal adult rat brain. Thus, immunohistochemical labeling for citrullinated epitopes was seen in white matter areas consistent with myelin staining; however, in general, it was more prominent and uniform in the caudal CNS (spinal cord, medulla oblongata, pons, and cerebellum) than in more rostral areas. F95 staining was also seen in cells and fibers often intimately associated with blood vessels and/or ventricular surfaces. By using dual-color immunofluorescence, the vast majority of this latter staining was colocalized within a subset of astrocytes also immunoreactive for glial fibrillary acidic protein (GFAP). By using Western blot analysis of rat brain proteins, multiple GFAP- and MBP-immunoreactive proteins and peptide fragments were seen, and many of them were also reactive with the F95 antibody. Thus, the present study not only demonstrates that citrullinated epitopes in normal rat brain are most concentrated in subsets of myelin and astrocytes but also provides evidence that GFAP, like MBP, may be present as multiple citrullinated isoforms. PMID- 12655509 TI - Neuron-specific distribution of P2X7 purinergic receptors in the monkey retina. AB - Extracellular ATP is a signaling molecule, working through P2X purinoceptors in the nervous system. P2X7 is a major subtype of the purinoceptors in the brain, where it is expressed mostly in glia cells and considered to work as a trigger of cytolysis. In the rodent retina, however, P2X7 is expressed in several classes of neurons including ganglion cells. In the present study we identified cells immunopositive for P2X7 by double immunolabeling. Immunoreactivity for P2X7 was observed in the inner nuclear layer (INL), the inner plexiform layer (IPL), and the ganglion cell layer (GCL). In the INL, strongly immunopositive cells corresponded to the subpopulation of horizontal cells. In the IPL, fine processes were immunopositive. In the GCL, most of the ganglion cells showed P2X7 immunoreactivity. At the ultrastructural level, immunoreactivity was confirmed in the cytoplasm of ganglion cells. No P2X7 immunoreactivity was found in non-neural cells, i.e., Muller cells or microglia. The immunohistochemical distribution of other purinoceptor subtypes (P2X1, P2X2, and P2X4) was also examined in the monkey retina. Immunoreactivity for P2X1 was strongly detected in a band, in sublamina a of the IPL. The band existed at almost the same level as tyrosine hydroxylase immunoreactivity, but did not seem to actually overlap. P2X2 was not expressed in the retina, and P2X4 was only faintly expressed at the scleral margin of the INL. Because P2X7 in the primate retina is expressed exclusively in neurons, it may in this location be involved in neural transmission rather than in cytolysis, as found for glia cells. PMID- 12655511 TI - GABA and glutamate in mating-activated cells in the preoptic area and medial amygdala of male gerbils. AB - The posterodorsal medial amygdala (MeApd), the posterodorsal preoptic nucleus (PdPN), and the medial cell group of the sexually dimorphic preoptic area (mSDA) contain cells that are activated specifically at ejaculation as assessed by Fos expression. The mSDA also expresses Fos early in the mating context. Because little is known about the neurotransmitters of these activated cells, the possibility that they use gamma-aminobutyric acid (GABA) or glutamate was assessed. Putative glutamatergic cells were visualized with immunocytochemistry (ICC) for glutamate and its neuron-specific transporter. Their distributions were compared with those of GABAergic cells visualized with ICC for the 67-kDa form of glutamic acid decarboxylase (GAD(67)) and in situ hybridization for GAD(67) messenger RNA (mRNA). Colocalization of Fos and GAD(67) mRNA in recently mated males indicated that half of the activated cells in the PdPN, mSDA, and lateral MeApd are GABAergic. Colocalization of Fos and glutamate suggested that a quarter of the activated mSDA and lateral MeApd cells are glutamatergic. The PdPN does not appear to have glutamatergic cells. In the lateral MeApd, the percentage of activated cells that are GABAergic (45%) matches the percentage that project to the principal part of the bed nucleus of the stria terminalis (BST; 43%), and the percentage likely to be glutamatergic (27%) matches the percentage projecting to the mSDA (27%). The latter could help to trigger ejaculation. The distribution of GABAergic and putative glutamatergic cells in the caudal preoptic area, caudal BST, and medial amygdala of male gerbils is also described. PMID- 12655510 TI - Neurogranin expression by cerebellar neurons in rodents and non-human primates. AB - Neurogranin (NG) is a brain-specific protein kinase C substrate involved in the regulation of calcium signaling and neuronal plasticity. A rostrocaudal expression profile, with large amounts in telencephalic brain regions and low expression levels in phylogenetically older brain structures, was reported previously. In the cerebellum, expression of NG has not been described. By using immunocytochemistry and in situ hybridization, we found that NG is expressed in the mouse (C57Bl/6), rat (Wistar), and monkey (Cercopithecus aetiops) cerebella. In the mouse cerebellum, Golgi cells were strongly immunoreactive for NG, whereas other cerebellar neurons were devoid of this protein. Cell counts showed 1.6-fold more immunopositive Golgi cells in the hemispheres (61.1 +/- 8.0 cells/mm(2)) than in the vermis (37.5 +/- 3.3 cells/mm(2)). Developmental studies showed detectable NG in the mouse cerebellum as early as on postnatal day 10 (P10). In contrast to the mouse, in the rat cerebellum we found only a few Golgi cells containing NG (hemispheres, 2.4 +/- 0.5 cells/mm(2); vermis, 1.5 +/- 0.3 cells/mm(2)). In the monkey cerebellum, unipolar brush cells, localized in the granular layer, were heavily labeled, whereas Golgi cells were devoid of NG. This study demonstrated that NG is strongly expressed in specific gamma-aminobutyric acidergic neurons in the rodent cerebellum. In addition, NG expression in the primate cerebellum by brush cells, which are excitatory, showed remarkable cell type-specific and species-specific expression patterns of a postsynaptic protein mediating calcium signaling mechanisms. PMID- 12655512 TI - Efferent connections of the dorsomedial thalamic nuclei of the domestic chick (Gallus domesticus). AB - Small iontophoretic injections of the anterograde tracer Phaseolus vulgaris leucoagglutinin were placed in the thalamic anterior dorsomedial nucleus (DMA) of domestic chicks. The projections of the DMA covered the rostrobasal forebrain, ventral paleostriatum, nucleus accumbens, septal nuclei, Wulst, hyperstriatum ventrale, neostriatal areas, archistriatal subdivisions, dorsolateral corticoid area, numerous hypothalamic nuclei, and dorsal thalamic nuclei. The rostral DMA projects preferentially on the hypothalamus, whereas the caudal part is connected mainly to the dorsal thalamus. The DMA is also connected to the periaqueductal gray, deep tectum opticum, intercollicular nucleus, ventral tegmental area, substantia nigra, locus coeruleus, dorsal lateral mesencephalic nucleus, lateral reticular formation, nucleus papillioformis, and vestibular and cranial nerve nuclei. This pattern of connectivity is likely to reflect an important role of the avian DMA in the regulation of attention and arousal, memory formation, fear responses, affective components of pain, and hormonally mediated behaviors. PMID- 12655514 TI - Decreased immunoreactivity for p27 protein in patients with early-stage breast carcinoma is correlated with HER-2/neu overexpression and with benefit from one course of perioperative chemotherapy in patients with negative lymph node status: results from International Breast Cancer Study Group Trial V. AB - BACKGROUND: The objective of this study was to clarify the prognostic and predictive value of immunoreactivity for the cyclin-dependent kinase inhibitor p27(Kip1) in patients with early-stage breast carcinoma and to investigate its relation with clinicopathologic features and other markers. METHODS: Immunoreactivity for p27 protein was analyzed on tumor slides from 461 patients who were enrolled in the International Breast Cancer Study Group (IBCSG) Trial V (median follow-up, 13 years), including 198 patients with lymph node negative disease and 263 patients with lymph node positive disease. Tumors with < 50% immunoreactive neoplastic cells were considered low expressors. Immunoreactivity for p27 was correlated with several clinicopathologic characteristics. Disease free survival (DFS) and overall survival were analyzed according to p27 immunoreactivity and treatment group. RESULTS: In the lymph node negative population, decreased p27 immunoreactivity was associated with higher tumor grade (P = 0.001) and HER-2/neu overexpression (P = 0.04). In the lymph node positive population, low p27 expression was associated with higher tumor grade (P = 0.01), low expression of thymidylate synthase (P = 0.001), and higher Ki-67 expression (P = 0.007). DFS was not significantly different according to p27 status in either lymph node negative patients (10-year DFS: low p27 expression, 53% +/- 5%; high p27 expression, 55% +/- 5%) or in lymph node positive patients (10 year DFS: low p27 expression, 33% +/- 4%; high p27 expression, 32% +/- 4%). However, in the lymph node negative population, the benefit of one course of perioperative chemotherapy with cyclophosphamide, methotrexate, and 5-fluorouracil was confined exclusively to patients with tumors that showed reduced p27 immunoreactivity (P = 0.03; test for interaction). CONCLUSIONS: This analysis indicates that p27 immunoreactivity has little if any prognostic value in patients with early-stage breast carcinoma. However, these findings suggest that, in patients with breast carcinoma who have negative lymph node status, reduced p27 immunoreactivity is associated with HER-2/neu overexpression and may be predictive of a benefit from the early administration of adjuvant chemotherapy. PMID- 12655515 TI - Epithelial lesions in prophylactic mastectomy specimens from women with BRCA mutations. AB - BACKGROUND: It has been suggested that BRCA-associated breast carcinoma may often lack a detectable preinvasive phase. To investigate this hypothesis, the authors compared the prevalence of histopathologic lesions in prophylactic mastectomy (PM) specimens from women with BRCA mutations and in mastectomy specimens obtained at autopsy from an age and race-matched comparison group without a known cancer predisposition. METHODS: All specimens from women with a deleterious BRCA1 or BRCA2 mutation who participated in an ongoing follow-up study and underwent PM at Memorial Sloan-Kettering Cancer Center between November 1, 1987 and May 31, 2001 were reviewed. For each case, breast tissue from two age and race-matched women without a known cancer predisposition was also reviewed. The prevalence of benign, premalignant, and cancerous lesions was compared. RESULTS: Mastectomy specimens from 24 cases and 48 comparison subjects were reviewed. Ductal carcinoma in situ (DCIS), atypical ductal hyperplasia (ADH), and atypical lobular hyperplasia (ALH) were all more common in PM specimens from women with BRCA mutations than in those from the comparison group. The odds ratio for the detection of any high-risk lesion (DCIS, lobular carcinoma in situ, ADH, or ALH) in specimens from BRCA mutation carriers was 12.7 (95% confidence interval, 3.1 52.4; P < 0.001). CONCLUSIONS: Lesions associated with an increased risk of subsequent malignancy are more common in PM specimens from women with BRCA mutations than in breast tissue obtained at autopsy from unaffected women without a known predisposition. This finding suggests that hereditary breast carcinoma has a preinvasive phase that may be detectable with aggressive surveillance. PMID- 12655516 TI - Carcinoid heart disease: the role of urinary 5-hydroxyindoleacetic acid excretion and plasma levels of atrial natriuretic peptide, transforming growth factor-beta and fibroblast growth factor. AB - BACKGROUND: Serotonin excretion plays a role in the development of carcinoid heart disease (CHD), but the exact pathogenesis is not known. In the current study, the authors evaluated 24-hour urinary 5-hydroxyindoleacetic acid (5-HIAA) excretion, as well as plasma levels of transforming growth factor-beta (TGF beta), fibroblast growth factor (FGF), and atrial natriuretic peptide (ANP) in patients with and without CHD determined by ultrasound examination. METHODS: Urine and plasma samples were obtained for 37 patients and cardiac ultrasound was performed during follow-up in 1999 and 2000. Median 5-HIAA excretion was calculated for the period between diagnosis and ultrasound examination. CHD was defined as the thickening of the tricuspid valve with additional III-IV/IV tricuspid valve regurgitation. RESULTS: CHD was found in 9 of 37 patients (24%). No significant differences were found for age, gender, presence, and duration of liver metastases. All CHD patients had symptoms of the carcinoid syndrome compared with 71% of the non-CHD patients (P = 0.159). Median 5-HIAA excretion was significantly higher in the CHD group compared with the non-CHD group: 576 micromol/24 hours versus 233 micromol/24 hours (P = 0.02). No difference in TGF beta and FGF plasma levels was observed between both groups (P = 0.139 and P = 0.985, respectively), nor was there a correlation with morphology of the tricuspid valve or degree of dilatation of the right atrium/ventricle. However, the CHD group had higher median ANP levels than the non-CHD group: 48 ng/L and 25 ng/L, respectively (P = 0.026). CONCLUSIONS: High levels of 5-HIAA excretion and plasma ANP were found to be associated with CHD. No significant relation with TGF beta or FGF was been found. PMID- 12655517 TI - Pattern of recurrence following complete resection of esophageal carcinoma and factors predictive of recurrent disease. AB - BACKGROUND: Despite increasingly radical surgery for esophageal carcinoma, a large number of patients still experience recurrent disease soon after operation. The current study was undertaken to evaluate the pattern of recurrence after curative esophagectomy for cancer of the thoracic esophagus and to identify factors predictive of recurrent disease. METHODS: A total of 439 consecutive patients discharged from the authors' institution following R0 resection between January 1982 and July 2002 were followed for evidence of recurrence over a mean interval of 37.3 (range, 1-207) months. RESULTS: Overall 1-, 3- and 5-years survival rates were 91%, 54%, and 41%, respectively. Some 230 patients (52.4%) developed proven recurrence, of whom 24 were alive and 206 were dead at the time of writing. The median time to recurrence was 12.0 (range, 6-96) months, with a median survival thereafter of 7.0 (range, 0-83) months. The pattern of recurrence was local in 12.1%, regional in 20.5% (cervical 3.6%, mediastinal 14.8%, and abdominal 2.1%), and distant in 19.8%, respectively. The overall pattern of dissemination was significantly different according to the histologic subtype (P = 0.021). One hundred five (45.7%) of all recurrences occurred within 12 months of surgery, with local, regional, and distant recurrence occurring at a median of 14.0 (range, 6-77), 13.5 (range, 6-73), and 11.0 (range, 6-96) months, respectively; A factor predictive of recurrent disease was histologic tumor depth invasion (P = 0.001). CONCLUSIONS: Depth of tumor invasion should be used to identify patients who will have recurrence within 12 months of operation, so that these patients may be either entered into trials of multimodality treatment or offered nonsurgical palliation. PMID- 12655518 TI - Sperm banking and rate of assisted reproduction treatment: insights from a 15 year cryopreservation program for male cancer patients. AB - BACKGROUND: Semen cryostorage remains the only proven method to preserve fertility in men with cancer. However, the necessity and the justification of banking spermatozoa have been questioned because it has been reported that only a small percentage of patients are referred for assisted reproductive technologies using frozen semen. METHODS: A 15-year cryopreservation program involving 776 men with malignant diseases who were referred to the study institution for semen cryopreservation before proceeding with chemotherapy and/or radiotherapy was reviewed. Data regarding cancer diagnosis, age, semen quality, and use of frozen semen were analyzed. RESULTS: Sperm banking was not performed for 90 of the 776 subjects (11.6%) because of azoospermia. Sperm quality was reduced in men with testicular carcinoma. To date, 36 of the 686 patients who had banked spermatozoa (5.2%) have attended the clinic for assisted conception treatments using frozen spermatozoa. Cumulative rates related to the use of frozen semen at 4, 8, and 12 years were 4.5%, 8.7%, and 11.8%, respectively. Relevant factors demonstrated to be significantly associated with a lower probability of using frozen material were a younger age at the time of cryostorage and a diagnosis of testicular carcinoma. CONCLUSIONS: The rate of referral for assisted reproductive techniques in patients who have banked their semen is low but tends to rapidly increase extending the length of follow-up. The cumulative percentage of use is at least above 10% but a longer follow-up is required to draw definitive conclusions. A diagnosis of testicular carcinoma is associated with a lower rate of use when compared with other cancers. Cancer 2003;97:1624-9. PMID- 12655520 TI - Anatomic distribution of periprostatic adipose tissue: a mapping study of 100 radical prostatectomy specimens. AB - BACKGROUND: Because the prostatic capsule is often indistinct, recognition of extraprostatic extension by carcinoma depends heavily on the identification of carcinoma cells in the periprostatic adipose tissue. However, the distribution of adipose tissue along the prostatic surfaces has not been studied extensively. METHODS: The authors analyzed the periprostatic adipose tissue in specimens from 100 patients treated with radical retropubic prostatectomy and bilateral pelvic lymphadenectomy for prostatecarcinoma. Each specimen was totally embedded and examined by the whole mount method. The presence or absence of adipose tissue on the anterior, posterior, right, and left surfaces of the prostate was determined and analyzed. RESULTS: Periprostatic adipose tissue was present on 48% of all prostatic surfaces examined. The distribution of periprostatic adipose tissue varied among the different surfaces of the prostate, with the anterior, posterior, right, and left surfaces showing 44%, 36%, 59%, and 57% adipose tissue, respectively. The amount of periprostatic adipose tissue was similar among specimens from operations performed by different surgeons (P = 0.72). However, nerve-sparing procedures resulted in less adipose tissue (46%) than non nerve-sparing procedures (54%) (P = 0.01). CONCLUSIONS: Less than half of the prostatic surfaces examined were covered by adipose tissue. The absence of adipose tissue over large areas of the prostatic surface, especially the posterior surface, makes the evaluation of extraprostatic extension difficult and unreliable. Further refinement of the prostate carcinoma staging system is needed. PMID- 12655519 TI - Molecular markers of outcome after radiotherapy in patients with prostate carcinoma: Ki-67, bcl-2, bax, and bcl-x. AB - BACKGROUND: Abnormal expression of key proteins of the apoptotic pathway has been associated with poor prognosis, although there have been few studies of these correlations in patients with prostate carcinoma who are treated with radiotherapy. The current study examined the association between expression levels of Ki-67, bcl-2, bax, and bcl-x in pretreatment biopsy specimens and patient outcome after definitive radiotherapy alone. METHODS: Archival pretreatment prostate biopsy tumor tissue was retrieved from 106 patients with Stage T1-T3 prostate carcinoma who were treated at the University of Texas M. D. Anderson Cancer Center with external beam radiotherapy between 1987 and 1993. Expression levels of Ki-67 (MIB-1 staining; n = 106 patients), bcl-2 (n = 77 patients), bax (n = 70 patients), and bcl-x (both long and short splice variants; n = 72 patients) were determined by immunohistochemical staining. The Ki-67 labeling index (Ki67-LI) was available for all patients and was derived from the percentage of Ki-67 positive cells. Biochemical failure after radiotherapy was defined as three consecutive rises in prostate specific antigen level on follow up. The median follow-up was 62 months. RESULTS: High Ki67-LI (> 3.5%) expression was observed in 33% of patients, overexpression of bcl-2 was observed in 16% of patients, altered bax expression was observed in 23% of patients, and altered bcl x expression was observed in 53% of patients. There was no correlation found between the biomarkers. Kaplan-Meier survival estimates of freedom from biochemical failure (bNED) and the log-rank test revealed significantly lower rates in association with high Ki67-LI, positive bcl-2, and altered bax staining. No correlation was observed between bcl-x staining and bNED. Cox proportional hazards multivariate analysis confirmed that bcl-2 and bax were independent of pretreatment PSA level, Gleason score, disease stage, and Ki67-LI in predicting bNED. CONCLUSIONS: Abnormalities in the expression levels of bcl-2 and bax were associated with increased failure after patients were treated for prostate carcinoma with external beam radiotherapy. These biomarkers appeared to be useful in categorizing patient risk further, beyond Ki-67 staining and conventional clinical prognostic factors. PMID- 12655521 TI - Can patient selection for bladder preservation be based on response to chemotherapy? AB - BACKGROUND: Neoadjuvant chemotherapy for patients with muscle-invasive bladder carcinoma is given to treat micrometastases and to preserve the bladder. The objective of this study was to evaluate the possibility of bladder preservation in patients with muscle-invasive bladder carcinoma who were treated with neoadjuvant methotrexate, vinblastine, doxorubicin, and cisplatin (M-VAC) chemotherapy. METHODS: One hundred four consecutive patients with T2-T4,N0,M0 transitional cell carcinoma of the bladder were treated with 3 cycles of neoadjuvant M-VAC chemotherapy. After clinical restaging, 52 patients underwent transurethral resection of the bladder (TURB) alone, 13 patients underwent partial cystectomy, and 39 patients underwent radical cystectomy. RESULTS: The median survival for the entire group was 7.49 years (95% confidence interval, 4.86-10.0 years). Forty-nine patients (49%) were T0 at the time of TURB after receiving M-VAC. Thirty-one of 52 patients (60%) who received chemotherapy and underwent TURB alone were alive at a median follow-up of 56 + months (range, 10 160 + months): Twenty-three patients (44%) in that TURB group maintained an intact bladder. Of 13 responding patients with monofocal lesions who underwent partial cystectomy, only 1 patient required salvage cystectomy, and survival generally was good. The 5-year survival rate for this group was 69%. With a long median follow-up of 88 + months (range, 16-158 months), 4 patients (31%) were alive with a functioning bladder. In the radical cystectomy group, the median follow-up was 45 months (range, 4-172 + months), and 15 of 39 patients (38%) patients remained alive. In 77 patients who had their tumors down-staged to T0 or superficial disease, the median follow-up was 63 months (range, 4-172 + months), and the 5-year rate survival was 69%. This is in contrast to a 5-year survival rate of only 26% in 27 patients who failed to respond and had a status >/= T2 after receiving chemotherapy (median follow-up, 31 months; range, 7-156 + months). The median survival for 27 elderly patients (age >/= 70 years; median age, 73 years; range, 70-82 years) was 90 months (7.5 years). For elderly patients who underwent TURB and partial cystectomy, the 5-year survival rate was 67% with a 109-month (9-year) median survival; 47% of patients preserved their bladders intact. The median follow-up of the living elderly patients was 61 months (range, 20-120 + months). CONCLUSIONS: Bladder sparing in selected patients on the basis of response to neoadjuvant chemotherapy is a feasible approach that should be confirmed in prospective, randomized trials. Selected elderly patients are candidates for this approach. PMID- 12655522 TI - Patient satisfaction with treatment decisions for clinically localized prostate carcinoma. Results from the Prostate Cancer Outcomes Study. AB - BACKGROUND: Men with early-stage prostate carcinoma can choose aggressive treatment or conservative management. The authors used data from a population based cohort of men with clinically localized prostate carcinoma to describe satisfaction with treatment decisions 24 months after diagnosis and to examine the association of demographic, socioeconomic, and clinical characteristics with treatment satisfaction. METHODS: The authors evaluated 2365 men with clinically localized prostate carcinoma, diagnosed between October 1994 and October 1995, in one of six Surveillance, Epidemiology, and End Results tumor registries and who were available for 24 month follow-up. Medical record review and patient completed surveys provided demographic, socioeconomic, and clinical data and measured treatment satisfaction. Logistic regression analyses were used to identify factors associated with satisfaction. RESULTS: Overall, 59.2% of subjects were delighted or very pleased with their treatment selection. The perception of being cancer free (66.4%), maintaining urinary (64.2%) and bowel (60.5%) control and normal erectile function (65.9%), having good general health (71.3%), and preserving social relationships (68.1%) were significantly associated with being satisfied (P < 0.05). Men receiving no active treatment were less satisfied (50.5%, P < 0.001) than actively treated men, and Hispanic men were less satisfied than non-Hispanic white men after undergoing radical prostatectomy (50.1% vs. 58.0%, P = 0.05) or androgen deprivation (29.7% vs. 71.8%, P < 0.02). CONCLUSIONS: The majority of men were satisfied with their treatment selection for clinically localized prostate carcinoma. Receiving an active treatment, believing oneself to be free of cancer, avoiding treatment complications, and having good overall health and social support were positively associated with satisfaction. PMID- 12655523 TI - Prediction of progression after radical nephrectomy for patients with clear cell renal cell carcinoma: a stratification tool for prospective clinical trials. AB - BACKGROUND: The objective of the current study was to develop an algorithm to predict progression to metastases after radical nephrectomy for patients with clinically localized renal cell carcinoma (RCC) to allow stratification of patients for potential adjuvant therapy trials. METHODS: The authors identified 1671 sporadic patients with clinically localized, unilateral clear cell RCC who underwent radical nephrectomy between 1970 and 2000. The clinical features examined included age, gender, smoking history, recent onset hypertension, performance status, and presenting symptoms. The pathologic features examined included surgical margins, tumor stage, regional lymph node status, tumor size, nuclear grade, histologic tumor necrosis, sarcomatoid component, cystic architecture, and multifocality. Metastases free survival was estimated using the Kaplan-Meier method. A multivariate Cox proportional hazards regression model was fit to determine associations between the clinical and pathologic features and distant metastases. RESULTS: The median follow-up was 5.4 years (range, 0-31 years). Metastases occurred in 479 patients at a median of 1.3 years (range, 0-25 years) after nephrectomy. The estimated metastases free survival rates were 86.9% at 1 year, 77.8% at 3 years, 74.1% at 5 years, 70.8% at 7 years, and 67.1% at 10 years. Multivariate analysis showed that the following features were associated with progression to metastases: tumor stage, regional lymph node status, tumor size, nuclear grade, and histologic tumor necrosis (P < 0.001 for all). CONCLUSIONS: In patients with clear cell RCC, tumor stage, regional lymph node status, tumor size, nuclear grade, and histologic tumor necrosis showed statistically significant associations with progression to metastatic RCC. The authors present a scoring algorithm based on these features that can be used to predict disease progression after patients undergo radical nephrectomy for clinically localized clear cell RCC. Cancer 2003;97:1663-71. PMID- 12655524 TI - Correlation of cervical carcinoma and precancerous lesions with human papillomavirus (HPV) genotypes detected with the HPV DNA chip microarray method. AB - BACKGROUND: Human papillomavirus (HPV) infection is considered to play an important role in the development of cervical carcinoma, and it is known that certain HPV types, such as HPV-16 and HPV-18, are highly associated with cervical carcinoma. However, the pathologic behavior of other HPV types remains unclear. Recently, a new HPV detection technique, the HPV DNA chip, was introduced. The HPV DNA chip harbors 22 HPV probes and has the advantage of being able to detect 22 HPV types simultaneously. To evaluate the quality of the HPV DNA chip method and to identify HPV types related to cervical carcinoma and precancerous lesions, the authors performed HPV typing in cervical specimens from 1983 patients and compared their cytologic and histologic diagnoses. METHODS: The HPV DNA chip was used for HPV typing. Among 1983 patients who were tested for HPV types, cervical smear cytology was performed in 1650 patients, and 677 of those patients underwent cervical biopsy. RESULTS: Among the 1650 smears that were examined cytologically, 92.7% (114 of 123 smears) of low-grade squamous intraepithelial lesions (LSILs), 98.1% (106 of 108 smears) of high-grade squamous intraepithelial lesions (HSILs), and 96.3% (51 of 53 smears) of carcinomas were HPV positive, compared with only 35.1% of smears with normal cytology that were HPV positive. HPV-16 was the most prevalent type (chi-square test; P < 0.01) in LSILs (28.5%), in HSILs (51.9%), and in carcinomas (62.5%) followed by HPV-58 and a group of low risk types (HPV-6, HPV-11, HPV-34, HPV-40, HPV-42, HPV-43,and HPV-44) in LSILs. HPV-58 (15.7%), HPV-18 (6.7%), and HPV-52 (4.6%) were the next most prevalent types after HPV-16 in HSILs. HPV-18 (11.4%) and HPV-58 (11.4%) were the second most common types in carcinomas. HPV-58 had the highest positive predictive value (54.9%) for the detection of histologically confirmed HSIL or carcinoma, whereas HPV 16 had the highest negative predictive value (80.6%). The sensitivity (96.0%) of the HPV test using the DNA chip method for detecting HSIL or carcinoma was superior compared with the sensitivity of cytologic diagnosis (83.6%). CONCLUSIONS: The HPV DNA chip provides a very sensitive method for detecting 22 HPV genotypes with reasonable sensitivity (96.0%) and reasonable negative predictive value (96.9%), and it overcomes the low sensitivity of cytologic screening for the detection of HSIL or carcinoma. HPV-58, HPV-52, and HPV-56, as well as HPV-16 and HPV-18, were associated highly with HSIL and carcinoma in the current large series. In addition, multiple HPV infection was associated less frequently with cervical carcinoma and with precancerous lesions compared with normal cytology. PMID- 12655525 TI - Spectroscopic detection and evaluation of morphologic and biochemical changes in early human oral carcinoma. AB - BACKGROUND: Understanding the development and progression of head and neck squamous cell carcinoma is key in the quest for the early diagnosis and prevention of this type of malignancy. The current study correlated early biochemical and histologic changes in oral tissue with spectral features in fluorescence, reflectance, and light scattering spectra acquired in vivo to diagnose early stages of oral malignancies. METHODS: A total of 91 tissue sites from 15 patients with varying degrees of malignancy (normal, dysplastic, and cancerous sites) and 8 healthy volunteers were analyzed with 3 spectroscopic techniques. Direct biochemical information regarding oral tissue native fluorophores was obtained with intrinsic fluorescence spectroscopy by fitting a linear combination of collagen and the reduced form of nicotinamide adenine dinucleotide (NADH) fluorescence spectra to the intrinsic tissue fluorescence spectra excited with 337 nanometer (nm) and 358-nm laser light. Diffuse reflectance spectroscopy was used to provide information regarding tissue absorption and structure, such as hemoglobin concentration and stroma density, by measuring the wavelength-dependent absorption and scattering coefficients. By subtracting the diffusely reflected component from the measured reflectance, light scattering spectroscopy (LSS) information resulting from single backscattering from epithelial cell nuclei was obtained. LSS provides information concerning the size distribution of cell nuclei. RESULTS: These optically extracted tissue parameters provide biochemical or structural information in vivo without the need for tissue excision, and can be used to diagnose tissue abnormalities. By combining the information provided by the three techniques, a method known as trimodal spectroscopy, a sensitivity and specificity of 96% and 96%, respectively, in distinguishing cancerous/dysplastic (mild, moderate, and severe) from normal tissue was achieved. In addition, the authors were able to distinguish dysplastic from cancerous tissue with a sensitivity of 64% and a specificity of 90%. CONCLUSIONS: The results of the current study demonstrated that Trimodal spectroscopy is a highly sensitive and specific technique with which to diagnose tissue abnormalities. PMID- 12655526 TI - Stricture of the proximal esophagus in head and neck carcinoma patients after radiotherapy. AB - BACKGROUND: It is well recognized that many patients with head and neck carcinoma have problems with food intake and malnutrition. The objective of the current study was to determine the clinical pattern of patients with nonneoplastic stricture of the upper esophagus after radiotherapy for head and neck carcinoma. METHODS: A retrospective chart study of 22 patients with stricture of the proximal esophagus diagnosed between 1993 and 1999 at Karolinska Hospital was performed. The dose volume histograms of the first 2 cm and 5 cm, respectively, of the proximal esophagus were calculated. RESULTS: Five of the patients (23%) had total obliteration. The first 2 cm of the esophagus received at least 60 grays (Gy) in > 80% of the volume. Radiation injury was not reported to occur at doses < 60 Gy. There was a correlation found between dysphagia during radiotherapy and the development of proximal esophageal stricture. Stricture was diagnosed 1-60 months (median, 6 months) after radiotherapy. In 18 patients, the stricture was treated with single or repeated endoscopic dilation. These treatments allowed a nearly normal diet in 78% of the patients. CONCLUSIONS: Stricture of the upper esophagus is one deglutition disorder that is reported to occur after radiotherapy for head and neck carcinoma. In the current study, the authors emphasize the importance of knowing the tolerance of the normal esophagus to irradiation as well as early diagnosis of stricture of the proximal esophagus because this condition may lead to physical and emotional distress. PMID- 12655527 TI - Defects in the opioid growth factor receptor in human squamous cell carcinoma of the head and neck. AB - BACKGROUND: The endogenous opioid peptide, [Met(5)]-enkephalin, termed opioid growth factor (OGF), interacts with its receptor (OGFr) to play a role as a constitutively expressed inhibitory growth factor in the proliferation of epithelial cells. This study compared protein and gene expression of OGFr in surgical specimens of human squamous cell carcinoma of the head and neck (SCCHN) with normal epithelium. METHODS: Tissues from 64 patients with SCCHN and from 49 patients undergoing uvulapalatoplasty or tonsillectomy were utilized. Binding affinity and capacity were assessed by receptor binding assays and the levels of OGFr protein were determined by quantitative Western blot analysis. Immunohistochemistry assessed the presence and distribution of OGFr. Levels of OGFr mRNA were quantitated by Northern blot analysis. Protein and gene expressions of OGFr also were evaluated in the margins of SCCHN. RESULTS: Binding analyses indicated nearly ninefold fewer OGFr binding sites in tumor tissue in comparison with normal samples. The OGFr protein levels were reduced fivefold in tumor tissues relative to normal epithelium. Values of OGFr mRNA were comparable in tumors and normal epithelium. Tumor margins had intermediate levels of protein and binding, but OGFr mRNA values were similar to those of normal specimens. CONCLUSIONS: These data demonstrate that OGFr is defective in SCCHN and that translation/posttranslation of OGFr protein, but not transcriptional levels of the OGFr gene, is involved. In addition, the attenuated levels of OGFr binding capacity may serve as a marker for SCCHN. These subnormal levels of OGFr may be responsible in part for tumor progression, diminishing the interaction of OGF with OGFr that aids in stabilizing cell replication by an inhibitory mechanism. Gene therapy to reinstate OGFr and/or function could provide a useful treatment for inhibiting tumor progression. PMID- 12655528 TI - Fractionated cyclophosphamide, vincristine, liposomal daunorubicin, and dexamethasone plus rituximab and granulocyte-macrophage-colony stimulating factor (GM-CSF) alternating with methotrexate and cytarabine plus rituximab and GM-CSF in patients with Richter syndrome or fludarabine-refractory chronic lymphocytic leukemia. AB - BACKGROUND: Therapy for patients with Richter syndrome (RS) or fludarabine refractory chronic lymphocytic leukemia (CLL) is unsatisfactory. A Phase II study was conducted to evaluate an alternating combination cytotoxic regimen given with rituximab and granulocyte-macrophage-colony stimulating factor (GM-CSF) in these patients. METHODS: Fludarabine-refractory CLL was defined as failure to respond to most recent prior fludarabine-containing regimen. Patients received up to six cycles of fractionated cyclophosphamide, vincristine, liposomal daunorubicin, and dexamethasone (hyper-CVXD) plus rituximab and GM-CSF alternating with methotrexate and cytarabine plus rituximab and GM-CSF. Response, toxicity, and survival data were compared with data from prior therapy with hyper-CVXD alone in this patient group. RESULTS: Forty-nine patients with RS (n = 30 patients) or refractory CLL (n = 19 patients) were treated on study. Nine patients (18%) achieved a complete remission, and 11 patients achieved a partial remission (22%), for an overall objective response (OR) rate of 41%. With a median follow up of 7.5 months and a maximum follow-up of 15.2 months, the 12-month failure free survival (FFS) rate was 27%, and the overall survival (OS) rate was 39%. Nine patients (18%) died during the first cycle of therapy, and two patients (4%) died during the second cycle. There were no significant differences between the rates of OR, OS, and FFS in the current study and those obtained with hyper-CVXD alone on a prior study. CONCLUSIONS: The study regimen had activity and significant toxicity in patients with RS or fludarabine-refractory CLL. It was not clearly better compared with hyper-CVXD alone in this patient population. PMID- 12655529 TI - Comparison of allogeneic stem cell transplantation, high-dose cytarabine, and autologous peripheral stem cell transplantation as postremission treatment in patients with de novo acute myelogenous leukemia. AB - BACKGROUND: Postremission therapy is critical in maintaining complete remission (CR) in patients with de novo acute myelogenous leukemia (AML). The aim of this trial was to compare allogeneic stem cell transplantation (SCT), high-dose cytarabine (ara-C; HiDAC), and autologous SCT as postremission therapy in patients with de novo AML. METHODS: One hundred twenty patients age /= T2 tumors, and 28% of patients had HER-2 positive tumors. Eighty-four percent of patients received four courses of FAC, 8% of patients received 3 courses of FAC, and the remaining 8% of patients received 5-6 courses of FAC. The clinical response rate (complete response [CR] and partial response [PR]) was 78%, the imaging response rate (CR and PR) was 64%, and 15% of patients had a good pathologic response, defined as a CR or minimal residual disease (tumor measuring < 1 cm in greatest dimension and negative lymph nodes). Concordance between the three methods of response assessment (clinical, pathologic, and imaging) was modest and was best between clinical assessment and imaging assessment (64% concordance). HER-2 status did not correlate with pathologic or clinical response (assessed by physical examination or imaging), although a nonsignificant trend was noted toward better response in patients with breast tumors that overexpressed HER-2. CONCLUSIONS: The authors found no significant correlation between HER-2 expression and clinical or pathologic response to neoadjuvant chemotherapy in patients with breast carcinoma. PMID- 12655532 TI - Physical activity levels before and after a diagnosis of breast carcinoma: the Health, Eating, Activity, and Lifestyle (HEAL) study. AB - BACKGROUND: Increased body weight at the time patients are diagnosed with breast carcinoma has been associated with an increased risk of recurrence and reduced survival. Weight gain also is common after diagnosis. Increasing physical activity (PA) after diagnosis may minimize these adverse outcomes. In this population-based study, the authors investigated whether PA levels after diagnosis declined from prediagnosis levels and whether any changes in PA varied by disease stage, adjuvant treatment, patient age, or body mass index (BMI) in 812 patients with incident breast carcinoma (from in situ to Stage IIIa). METHODS: Types of sports and household activities and their frequency and duration for the year prior to diagnosis and for the month prior to the interview (i.e., 4-12 months postdiagnosis) were assessed during a baseline interview. RESULTS: Patients decreased their total PA by an estimated 2.0 hours per week from prediagnosis to postdiagnosis, an 11% decrease (P < 0.05). Greater decreases in sports PA were observed among women who were treated with radiation and chemotherapy (50% decrease) compared with women who underwent surgery only (24% decrease) or who were treated with radiation only (23%; (P < 0.05). Greater decreases in sports PA were observed among obese patients (41% decrease) compared with patients of normal weight (24% decrease; P < 0.05). CONCLUSIONS: PA levels were reduced significantly after patients were diagnosed with breast carcinoma. Greater decreases in PA observed among heavier patients implied a potential for greater weight gain among women who already were overweight. Randomized, controlled trials are needed to evaluate how PA may improve the prognosis for patients with breast carcinoma. PMID- 12655534 TI - Clinical importance of molecular determinations in gynecologic patients with multiple tumors. AB - BACKGROUND: The prognosis and treatment of patients with multiple tumors may depend on the correlation between tumors: multiple primary tumors, or recurrent tumors, and metastatic disease. The authors investigated whether the detection of molecular aberrations in multiple gynecologic tumors in individual patients provided clinically useful information on the correlation between the tumors. METHODS: Between 1999 and 2001, molecular analyses were performed on tissue from 15 gynecologic patients, all with multiple tumors. The molecular analyses included loss of heterozygosity determinations at eight DNA loci and mutation analyses of p53 exons 5-8 using the single-strand conformation polymorphism method. Previously, it was not possible to use routine diagnostic histopathology to determine accurately the correlation between multiple lesions in patients with gynecologic malignancies, information that may have an impact on clinical decision-making and prognosis. RESULTS: Molecular results were obtained from all tumors from each of the 15 patients. The DNA alterations detected provided evidence that two patients had second primary tumors, nine patients had a single tumor with metastases, and four patients had two independent primary tumors as well as metastatic disease. The results provided additional diagnostic information and contributed to clinical decision-making. CONCLUSIONS: The authors demonstrated that, by comparing DNA alterations in multiple tumors within an individual patient, evidence about correlations between the tumors can be obtained. These investigations can be performed on routinely processed tissues, and the results may be of clinical importance in helping to determine the management or prognosis of patients with gynecologic malignancies. PMID- 12655535 TI - Outpatient, sequential, parenteral-oral antibiotic therapy for lower risk febrile neutropenia in children with malignant disease: a single-center, randomized, controlled trial in Argentina. AB - BACKGROUND: Recent reports and previous randomized trials conducted at the authors' institution suggested that children with lower risk febrile neutropenic (LRFN) may benefit from substitution of oral antibiotic therapy for parenteral therapy. The objective of this study was to determine the efficacy of parenteral oral outpatient therapy in the management of children with LRFN who were receiving treatment for malignant disease. METHODS: From August 2000 to April 2002, 135 children with a median age of 7.5 years (range, 1.6-15.8 years) who had a total of 177 episodes of LRFN were included in a prospective, randomized, single-institution trial. Children with LRFN received a single dose of ceftriaxone and amikacin and completed a risk-assessment work-up. All patients were discharged immediately and, at 24 hours, were allocated randomly to two groups: Group A (89 episodes) received oral ciprofloxacin, and Group B (88 episodes) received intravenous ceftriaxone. RESULTS: Most patients (61% in Group A and 51% in Group B) were receiving treatment for leukemia (P value not significant [NS]). Twenty-eight children (31%) in Group A and 22 children (25%) in Group B displayed unexplained fever (P value NS). No significant differences in sites of initial infection were found between the two groups. The median duration of neutropenia was 4.2 days and 4.7 days for Group A and Group B, respectively (P value NS); the median duration of fever was 2.3 days and 2.6 days, respectively (P value NS); and the median duration of antibiotic treatment was 4.5 days and 4.8 days, respectively (P value NS). The overall results of the study were excellent. Only four treatment failures in Group A (5%) and 6 treatment failures in Group B (7%) were observed. These patients were readmitted to the hospital and did well with appropriate treatment. CONCLUSIONS: In children with LRFN who are receiving treatment for malignant disease, outpatient oral ciprofloxacin after 24 hours of a single dose of intravenous ceftriaxone and amikacin was as safe and efficacious as parenteral ceftriaxone. Outpatient management and early antibiotic withdrawal were safe for both groups. PMID- 12655536 TI - Extended-field radiotherapy and high-dose-rate brachytherapy in carcinoma of the uterine cervix: clinical experience with and without concomitant chemotherapy. AB - BACKGROUND: It has long been recognized that many patients with locally advanced carcinoma of the cervix harbor occult paraaortic metastases. A randomized study demonstrated that elective paraaortic irradiation improved survival and reduced distant metastases. More recently, concomitant chemotherapy with pelvic irradiation has improved survival among patients with locally advanced carcinoma of the cervix. This has led to a reexamination of the role of extended-field irradiation. An important issue is the toxicity of concomitant chemotherapy and extended-field radiotherapy. The authors report a retrospective analysis of their experience with extended-field radiotherapy and high-dose-rate brachytherapy with or without concomitant chemotherapy. METHODS: The authors treated 54 women with biopsy-confirmed carcinoma of the cervix using extended-field radiotherapy and high-dose-rate brachytherapy with or without concomitant chemotherapy. The histology was squamous cell carcinoma in 49 patients (91%) and nonsquamous cell carcinoma in 5 patients (9%). The median size of the primary tumor was 7 cm (range, 3-10 cm). Each patient received 45 grays (Gy) of external beam radiotherapy to the pelvis and the paraaortic region, followed by a parametrial boost (9 Gy) in the patients with disease extension to the parametrium or the pelvic side wall(s). Each patient also underwent two applications of high-dose rate brachytherapy, 1 week apart. The median dose delivered to Point A from each application was 9 Gy. Forty-four of the 54 patients (81%) received concomitant chemotherapy (cisplatin, 20 mg/m(2)/day for 5 days) during the first and the fourth weeks of external beam radiotherapy, and once after the second high-dose rate application. Chemotherapy was not assigned randomly. RESULTS: One of the 10 patients (10%) treated without chemotherapy experienced acute toxicity, whereas 41 of 44 patients (93%) who received chemotherapy suffered from acute toxicity, including hematologic toxicity, gastrointestinal toxicity, and deep venous thrombosis. During a median follow-up period of 28 months (range, 12-70 months), 6 of the 54 patients have died (11%). The actuarial rate of local control at 3 years is 100% among the patients treated without chemotherapy, compared with 85% among those receiving chemotherapy. No one failed in the paraaortic region. The actuarial rates of freedom from distant metastases are 90% and 95% among the patients treated without and with chemotherapy, respectively. The actuarial incidence of late toxicity is 10% among the patients treated without chemotherapy and 6% among those receiving chemotherapy. CONCLUSIONS: The regimen of extended field radiotherapy with concomitant cisplatin and high-dose-rate brachytherapy produced substantial acute toxicity, but its long-term toxicity is low and the preliminary tumor control excellent, albeit with limited follow-up. Only prospective, randomized trials can evaluate whether these results are truly better than those of pelvic radiotherapy with concomitant chemotherapy, or those of other regimens of extended-field radiotherapy with concomitant chemotherapy. Cancer 2003;97:1781-8. PMID- 12655537 TI - Adjuvant irradiation for cervical lymph node metastases from melanoma. AB - BACKGROUND: The risk of regional disease recurrence after surgery alone for lymph node metastases from melanoma is well documented. The role of adjuvant irradiation remains controversial. METHODS: The medical records of 160 patients with cervical lymph node metastases from melanoma were reviewed retrospectively. Of these, 148 (93%) presented with clinically palpable lymph node metastases. All patients underwent surgery and radiation to a median dose of 30 grays (Gy) at 6 Gy per fraction delivered twice weekly. Surgical resection was either a selective neck dissection in 90 patients or local excision of the lymph node metastasis in 35 patients. Only 35 patients underwent a radical, modified radical, or functional neck dissection. RESULTS: At a median follow-up of 78 months, the actuarial local, regional, and locoregional control rates at 10 years were 94%, 94%, and 91%, respectively. Univariate analysis of patient, tumor, and treatment characteristics failed to reveal any association with the subsequent rate of local or regional control. The actuarial disease-specific (DSS), disease-free, and distant metastasis-free survival (DMFS) rates at 10 years were 48%, 42%, and 43%, respectively. Univariate and multivariate analyses revealed that patients with four or more involved lymph nodes had a significantly worse DSS and DMFS. Nine patients developed a treatment-related complication requiring medical management, resulting in a 5-year actuarial complication-free survival rate of 90%. CONCLUSIONS: Adjuvant radiotherapy resulted in a 10-year regional control rate of 94%. Complications for all patients were rare and manageable when they did occur. The authors recommend adjuvant irradiation for patients with extracapsular extension, lymph nodes measuring 3 cm in size or larger, the involvement of multiple lymph nodes, recurrent disease, or any patient having undergone a selective therapeutic neck dissection. PMID- 12655538 TI - Modulation of antitumor immune responses by hematopoietic cytokines. AB - BACKGROUND: Advances in immunotherapy for the treatment of patients with malignant disease have led to increasingly successful use of these methods in the clinical setting. This review presents findings from recent studies that have explored improved methods for the presentation of tumor-associated antigens and for the restoration of tumor specific immune responses using cytokine therapy. METHODS: A review of human clinical trial research on immune cytokines from 1995 (MEDLINE) to the present was conducted. Particular attention was focused on articles that reported results from Phase II or later clinical studies in patients with malignant disease. RESULTS: The defects in cellular immunity commonly seen in patients with malignancies often are expressed as tumor specific anergy. Reversing patient tolerance to tumor antigens may be accomplished by treatment with immunoregulatory cytokines, such as Flt-3 and granulocyte macrophage-colony stimulating factor, that mature and activate dendritic cells. Published clinical studies indicate that granulocyte-macrophage-colony stimulating factor stimulates antigen-presenting cells and has promising antitumor activity as an adjunct or as stand-alone therapy for patients with malignant disease, including leukemia, melanoma, breast carcinoma, prostate carcinoma, and renal cell carcinoma. CONCLUSIONS: Immune-modulating cytokines may be used alone or in combination with other treatments to help restore immune function, improve response to tumor-associated antigens, and reduce the toxic effects of standard antitumor therapies. The evolving understanding of how dendritic cells regulate immune responses and promising results from published studies of immune-enhancing cytokines in the treatment of patients with malignant disease support the conduct of randomized clinical trials to confirm the clinical benefit of these immunotherapeutic strategies. PMID- 12655539 TI - Luteinizing hormone receptor status and clinical, pathologic, and prognostic features in patients with breast carcinomas. AB - BACKGROUND: It has been established that pregnancy protects against breast carcinoma, and animal models have shown that human chorionic gonadotropin (hCG) mimics this effect by inhibiting the initiation and progression of experimental breast carcinoma. Luteinizing hormone (LH)/hCG receptors (LHR) have been characterized in several human breast carcinoma cell lines and in a limited number of breast carcinoma biopsy specimens. These observations led to the suggestion that hCG may be used as a means of prevention and possibly treatment in patients with breast carcinoma. METHODS: The authors used immunocytochemistry to analyze tumors from 160 patients who were followed for a median of 2539 days. Using a cut-off value of 18% immunolabeled cells in each tumor, 72% of tumors were identified as LHR positive. The LHR-positive tumors were found more frequently in premenopausal women, who had tumors with greater cell differentiation and positive estrogen receptor alpha status. Infiltrating lobular carcinomas were positive for LHR more frequently compared with infiltrating ductal carcinomas. There was no correlation between LHR status and lymph node invasion, tumor size, or progesterone receptor status. RESULTS: Patients with LHR positive tumors had a longer metastasis free survival, although the statistical significance was slight (P = 0.07), most likely due to the limited number of events in the patients studied. Conversely, there was no difference between patients with LHR-positive or LHR-negative tumors in the local recurrence free interval. CONCLUSIONS: LHR status seems to be related in part to the degree of differentiation in breast tumors, confirming experimental evidence of the effect of hCG on mammary tissue. The presence of LHR is a tumor characteristic that largely is independent of other clinical and pathologic tumor features. It may be of interest in the future to correlate the presence of LHR with a possible therapeutic response in individual patients to hCG. PMID- 12655541 TI - Dermoscopic and histopathologic diagnosis of equivocal melanocytic skin lesions: an interdisciplinary study on 107 cases. PMID- 12655542 TI - Focus on the molecular genetics of phenylketonuria. PMID- 12655543 TI - PAHdb 2003: what a locus-specific knowledgebase can do. AB - PAHdb, a legacy of and resource in genetics, is a relational locus-specific database (http://www.pahdb.mcgill.ca). It records and annotates both pathogenic alleles (n = 439, putative disease-causing) and benign alleles (n = 41, putative untranslated polymorphisms) at the human phenylalanine hydroxylase locus (symbol PAH). Human alleles named by nucleotide number (systematic names) and their trivial names receive unique identifier numbers. The annotated gDNA sequence for PAH is typical for mammalian genes. An annotated gDNA sequence is numbered so that cDNA and gDNA sites are interconvertable. A site map for PAHdb leads to a large array of secondary data (attributes): source of the allele (submitter, publication, or population); polymorphic haplotype background; and effect of the allele as predicted by molecular modeling on the phenylalanine hydroxylase enzyme (EC 1.14.16.1) or by in vitro expression analysis. The majority (63%) of the putative pathogenic PAH alleles are point mutations causing missense in translation of which few have a primary effect on PAH enzyme kinetics. Most apparently have a secondary effect on its function through misfolding, aggregation, and intracellular degradation of the protein. Some point mutations create new splice sites. A subset of primary PAH mutations that are tetrahydrobiopterin-responsive is highlighted on a Curators' Page. A clinical module describes the corresponding human clinical disorders (hyperphenylalaninemia [HPA] and phenylketonuria [PKU]), their inheritance, and their treatment. PAHdb contains data on the mouse gene (Pah) and on four orthologous mutant mouse models and their use (for example, in research on oral treatment of PKU with the enzyme phenylalanine ammonia lyase [EC 4.3.1.5]). PMID- 12655544 TI - Phenylketonuria mutations in Europe. AB - Phenylketonuria (PKU) is heterogeneous. More than 400 different mutations in the phenylalanine hydroxylase (PAH) gene have been identified. In a systematic review of the molecular genetics of PKU in Europe we identified 29 mutations that may be regarded as prevalent in European populations. Comprehensive regional data for these mutations were collated from all available studies. The spectrum of mutations found in individual regions results from a combination of factors including founder effect, range expansion and migration, genetic drift, and probably heterozygote advantage. Common mutations include R408W on a haplotype 2 background in Eastern Europe, IVS10-11G>A in the Mediterranean, IVS12+1G>A in Denmark and England, Y414C in Scandinavia, I65T in Western Europe, and R408W on haplotype 1 in the British Isles. Molecular data from mild hyperphenylalaninemia (MHP) patients are available from a number of countries, but it is currently not possible to calculate relative allele frequencies. The available data on PAH mutations are useful for the understanding of both the clinical features and the population genetics of PAH deficiency in Europe. PMID- 12655545 TI - How PAH gene mutations cause hyper-phenylalaninemia and why mechanism matters: insights from in vitro expression. AB - Mutations in the human PAH gene, which encodes phenylalanine hydroxylase are associated with varying degrees of hyperphenylalaninemia (HPA). The more severe of these manifest as a classic metabolic disease--phenylketonuria (PKU). In vitro expression analysis of PAH mutations has three major applications: 1) to confirm that a disease-associated mutation is genuinely pathogenic, 2) to assess the severity of a mutation's impact, and 3) to examine how a mutation exerts its deleterious effects on the PAH enzyme, that is, to elucidate the molecular mechanisms involved. Data on expression analysis of 81 PAH mutations in multiple in vitro systems is summarized in tabular form online at www.pahdb.mcgill.ca. A review of these findings points in particular to a prevalent general mechanism that appears to play a major role in the pathogenicity of many PAH mutations. Amino acid substitutions promote misfolding of the PAH protein monomer and/or oppose the correct assembly of monomers into the native tetrameric enzyme. The resulting structural aberrations trigger cellular defenses, provoking accelerated degradation of the abnormal protein. The intracellular steady-state levels of the mutant PAH enzyme are therefore reduced, leading to an overall decrease in phenylalanine hydroxylation within cells and thus to hyperphenylalaninemia. There is considerable scope for modulation of the enzymic and metabolic phenotypes by modification of the cellular handling--folding, assembly, and degradation--of the mutant PAH protein. This has major implications, both for our understanding of genotype-phenotype correlations and for the development of novel therapeutic approaches. PMID- 12655546 TI - Phenylketonuria: genotype-phenotype correlations based on expression analysis of structural and functional mutations in PAH. AB - When analyzed in the context of the phenylalanine hydroxylase (PAH) three dimensional structure, only a minority of the PKU mutations described world-wide affect catalytic residues. Consistent with these observations, recent data point to defective folding and subsequent aggregation/degradation as a predominant disease mechanism for several mutations. In this work, we use a combined approach of expression in eukaryotic cells at different temperatures and a prokaryotic system with co-expression of chaperonins to elucidate and confirm structural consequences for 18 PKU mutations. Three mutations are located in the amino terminal regulatory domain and 15 in the catalytic domain. Four mutations were found to abolish the specific activity in all conditions. Two are catalytic mutations (Y277D and E280K) and two are severe structural defects (IVS10-11G>A and L311P). All the remaining mutations (D59Y, I65T, E76G, P122Q, R158Q, G218V, R243Q, P244L, R252W, R261Q, A309V, R408Q, R408W, and Y414C) are folding defects causing reduced stability and accelerated degradation, although some of them probably affect residues involved in regulation. In these cases, we have demonstrated that the amount of mutant PAH protein and residual activity could be modulated by in vitro experimental conditions, and therefore the observed in vivo metabolic variation may be explained by interindividual variation in the quality control systems. The results derived provide an experimental framework to define the mutation severity relating genotype to phenotype. They also explain the observed inconsistencies for some mutations in patients with similar genotype and different phenotypes. PMID- 12655547 TI - Comparative multiplex dosage analysis detects whole exon deletions at the phenylalanine hydroxylase locus. AB - We have developed quantitative comparative multiplex dosage analysis to detect altered copy number of regions of the phenylalanine hydroxylase gene. Out of 41 alleles (4% of 1,010 PKU chromosomes) on which a mutation had not been characterized previously, this technique has highlighted two novel mutations: deletions of exon 5 and of exon 6 on a total of eight alleles. Restriction-enzyme digestion of genomic DNA and hybridization to an amplified segment of the phenylalanine hydroxylase (PAH) cDNA probe PAH247 established the size of the deletion in five individuals to be between 700 and 900 bases. We also report somatic mosaicism in the parent of an affected child previously shown to have a deletion spanning exons 5 and 6. Finally, we report a putative duplication of a region encompassing exon 6 in an affected individual. PMID- 12655548 TI - Genetic diversity within the R408W phenylketonuria mutation lineages in Europe. AB - The R408W phenylketonuria mutation in Europe has arisen by recurrent mutation in the human phenylalanine hydroxylase (PAH) locus and is associated with two major PAH haplotypes. R408W-2.3 exhibits a west-to-east cline of relative frequency reaching its maximum in the Balto-Slavic region, while R408W-1.8 exhibits an east to-west cline peaking in Connacht, the most westerly province of Ireland. Spatial autocorrelation analysis has demonstrated that the R408W-2.3 cline, like that of R408W-1.8, is consistent with a pattern likely to have been established by human dispersal. Genetic diversity within wild-type and R408W chromosomes in Europe was assessed through variable number tandem repeat (VNTR) nucleotide sequence variation and tetranucleotide short tandem repeat (STR) allelic associations. Wild-type VNTR-8 chromosomes exhibited two major cassette sequence organizations: (a1)5-b3-b2-c1 and (a1)5-b5-b2-c1. R408W-1.8 was predominantly associated with (a1)5-B5-B2-C1. Both wild-type vntr-3 and r408w-2.3 chromosomes exhibited a single invariant cassette sequence organization, a2-b2-c1. STR allele distributions associated with the cassette variants were consistent with greater diversity in the wild-type VNTR-8 lineage and were suggestive of different levels of diversity between R408W-1.8 and R408W-2.3. The finding of greater genetic diversity within the wild-type VNTR-8 lineage compared to VNTR-3 suggests that VNTR-8 may be older within the European population. However, in the absence of a more extensive STR data-set, no such conclusions are possible for the respective R408W mutant lineages. PMID- 12655549 TI - A role for overdominant selection in phenylketonuria? Evidence from molecular data. AB - To date, the reason is unknown for the high prevalence of phenylalanine hydroxylase (PAH) mutations causing phenylketonuria (PKU) in extant European populations. However, molecular genetic data generated over the last decade suggest that the concomitant excess of (unaffected) PKU carriers is at least in part the result of overdominant selection ("heterozygous advantage"). Such selection would have acted upon several different mutations in different historical populations. The exact nature of the underlying selective mechanism is unknown; its elucidation requires further investigation. PMID- 12655550 TI - The molecular basis of phenylketonuria in Lithuania. AB - We report the spectrum of phenylalanine hydroxylase (PAH) gene mutations in patients with phenylketonuria (PKU) residing in Lithuania. A total of 184 independent chromosomes was investigated. R408W mutation was first analysed through restriction enzyme digestion of exon 12. The remaining uncharacterised PKU chromosomes were analysed by scanning the whole coding sequence of PAH gene by multiplex 'broad range' denaturing gradient gel electrophoresis. Mutations were identified by fluorescent automated sequencing or by restriction enzyme digestion analysis if an abnormal DGGE pattern was recognised. 21 different mutations were identified for 175 PKU chromosomes, with a mutation detection rate of 95%. The most common ones were R408W (73.5% chromosomes) and R158Q (7.0% chromosomes) whereas the remaining mutations appeared to be rare (relative frequencies 0.5%-2%). The high mutation detection rate obtained is an evidence of the efficiency of PAH genetic testing achieved in Lithuania. Moreover, the definition of the PKU mutation profile in the Lithuanian population will allow to perform a genotype-phenotype correlation study thus making feasible genotyped based prediction of the biochemical phenotype in newborns with hyperphenylalaninemia. This may be useful for refining diagnosis and anticipating dietary requirements. PMID- 12655551 TI - The molecular basis of phenylketonuria in Latvia. AB - Characterization of the molecular basis of phenylketonuria (PKU) in Latvia has been accomplished through the analysis of 96 unrelated chromosomes from 50 Latvian PKU patients. Phenylalanine hydroxylase (PAH) gene mutations have been analyzed through a combined approach in which R158Q, R252W, R261Q, G272X, IVS10 11G>A and R408W mutations were first screened for by PCR or restriction generating PCR amplification of PAH gene exons 5, 7, 11 and 12 followed by digestion with the appropriate diagnostic enzyme. Subsequently 'broad range' denaturing gradient gel electrophoresis analysis of the 13 PAH gene exons has been used to study uncharacterized PKU chromosomes. A mutation detection rate of 98% was achieved. 12 different mutations were found, with the most frequent mutation, R408W, accounting for 76% of Latvian PKU alleles. Six mutations (R408W, E280K, R158Q, A104D, R261Q and P281L) represent 92% of PKU chromosomes. PAH VNTR and STR alleles have been also identified and minihaplotype associations with PKU mutations were also determined. PMID- 12655552 TI - The molecular basis of phenylalanine hydroxylase deficiency in Croatia. AB - We present the results of a comprehensive analysis of mutations, polymorphisms and haplotypes in the phenylalanine hydroxylase (PAH) gene in 39 Croatian families with phenylketonuria (PKU). A total of 21 disease-causing mutations was identified on 78 out of 79 independent chromosomes. The commonest mutation, R408W on haplotype 2 was found with a relative frequency of 37 %. P281L accounted for 11 %, R261Q and E390G each for 9 % of mutant chromosomes. There were three novel mutations: L249P (c.746T>C) in exon 7, IVS8+2T>C (c.912T>C) in intron 8, and F402L (c.1206T>G) in exon 12 of the PAH gene. Two known PKU mutations were found in cis on the same chromosome in one family, highlighting the need to perform full mutation scanning in recessive disease genes for molecular diagnosis even if two known mutations have been identified in a patient. This is the first comprehensive report on PKU mutations in southeastern Europe, adding to the growing bulk of molecular data for population genetic investigations. PMID- 12655553 TI - Mutational spectrum in German patients with phenylalanine hydroxylase deficiency. AB - We report on the spectrum and frequency of mutations in the phenylalanine hydroxylase (PAH) gene in 226 German families with PAH deficiency, most of them from Southern Germany. A total of 88 mutations were identified in 428 out of 438 mutant PAH alleles including one novel stop mutation L293X (c.878T>A). In three families, two phenylketonuria (PKU) mutations were found in cis, and in one family a de novo mutation was observed. A comparison of the results from Southern Germany with those of other parts of Western Germany showed no obvious local mutation clustering. In addition we studied the genotypic spectrum of 39 Turkish families with PAH deficiency. Twenty-three mutations were identified in 73 out of 75 Turkish chromosomes including two novel mutations: E280A (c.839A>G) in Exon 7 and IVS10-7C>A (c.1066-7C>A) in Intron 10. A new polymorphism IVS4+47C/T (c.441+47C>T) was found in mutant and normal PAH alleles. Screening of 170 German and 150 Turkish individuals without family history of PAH deficiency revealed 10 and 12 heterozygotes, respectively, a higher frequency of carriers than expected. A novel mutations of uncertain functional relevance, R169H (c.506G>A) in Exon 5 was found in two Turkish heterozygotes. Most of the Turkish heterozygotes carried mild mutations, indicating that mild forms of PAH deficiency may be more common in that population than previously recognised. PMID- 12655554 TI - Tetrahydrobiopterin sensitivity in German patients with mild phenylalanine hydroxylase deficiency. AB - We report the results of tetrahydrobiopterin (BH4) loading tests in 10 German patients with mild phenylketonuria. A significant decline of phenylalanine values after application of BH4 was observed in all but one patients. Molecular genetic analyses revealed a range of different PAH gene mutations. Re-testing of one patient previously reported as non-responsive to BH4 loading showed a moderate response with a higher dose of BH4. Nevertheless, there appear to be kinetic differences in phenylalanine hydroxylation in patients with the same genotype. Non-responsiveness to 20 mg/kg BH4 was observed only in a single patient who was compound heterozygous for the novel mutation R176P (c.527G>C) and the common null mutation P281L. In summary, our data are in line with recent reports indicating that BH4 sensitivity is a normal feature of most mild forms of PAH deficiency but may be influenced by other factors. PMID- 12655555 TI - Mutations in the AUH gene cause 3-methylglutaconic aciduria type I. AB - The conversion of 3-methylglutaconyl-CoA to 3-hydroxy-3-methylglutaryl-CoA is the only step in leucine catametabolism yet to be characterized at enzyme and DNA levels. The deficiency of the putative mitochondrial enzyme 3-methylglutaconyl CoA hydratase associates with the rare organic aciduria 3-methylglutaconic aciduria type I (MGA1), but neither the enzyme nor its gene have been described in any organism. Here we report that human 3-methylglutaconyl-CoA hydratase is identical with a previously described RNA-binding protein (designated AUH) possessing enoyl-CoA hydratase activity. Molecular analyses in five patients from four independent families revealed homozygosity or compound heterozygosity for mutations in the AUH gene; most mutations are predicted to completely abolish protein function. Mutations identified include c.80delG, R197X, IVS8-1G>A, A240V, and c.613_614insA. Clinical severity of MGA1 in published patients has been quite variable. Included in the present study is an additional patient with MGA1 who was detected by neonatal screening and has remained asymptomatic up to his present age of 2 years. The boy is homozygous for an N-terminal frameshift mutation in the AUH gene. Complete absence of 3-methylglutaconyl-CoA hydratase/AUH appears to be compatible with normal development in some cases. Further work is required to identify external or genetic factors associated with development of clinical problems in patients with MGA1. PMID- 12655556 TI - Human growth hormone 1 (GH1) gene expression: complex haplotype-dependent influence of polymorphic variation in the proximal promoter and locus control region. AB - The proximal promoter region of the human pituitary expressed growth hormone (GH1) gene is highly polymorphic, containing at least 15 single nucleotide polymorphisms (SNPs). This variation is manifest in 40 different haplotypes, the high diversity being explicable in terms of gene conversion, recurrent mutation, and selection. Functional analysis showed that 12 haplotypes were associated with a significantly reduced level of reporter gene expression whereas 10 haplotypes were associated with a significantly increased level. The former tend to be more prevalent in the general population than the latter (p<0.01), possibly as a consequence of selection. Although individual SNPs contributed to promoter strength in a highly interactive and non-additive fashion, haplotype partitioning was successful in identifying six SNPs as major determinants of GH1 gene expression. The prediction and functional testing of hitherto unobserved super maximal and sub-minimal promoter haplotypes was then used to test the efficacy of the haplotype partitioning approach. Electrophoretic mobility shift assays demonstrated that five SNP sites exhibit allele-specific protein binding. An association was noted between adult height and the mean in vitro expression value corresponding to an individual's GH1 promoter haplotype combination (p=0.028) although only 3.3% of the variance of adult height was found to be explicable by reference to this parameter. Three additional SNPs, identified within sites I and II of the upstream locus control region (LCR), were ascribed to three distinct LCR haplotypes. A series of LCR-GH1 proximal promoter constructs were used to demonstrate that 1) the LCR enhanced proximal promoter activity by up to 2.8-fold depending upon proximal promoter haplotype, and that 2) the activity of a given proximal promoter haplotype was also differentially enhanced by different LCR haplotypes. The genetic basis of inter-individual differences in GH1 gene expression thus appears to be extremely complex. PMID- 12655557 TI - Novel mutations of the growth hormone 1 (GH1) gene disclosed by modulation of the clinical selection criteria for individuals with short stature. AB - Subtle mutations in the growth hormone 1 (GH1) gene have been regarded as a comparatively rare cause of short stature. Such lesions were sought in a group of 41 individuals selected for short stature, reduced height velocity, and bone age delay; a group of 11 individuals with short stature and idiopathic growth hormone deficiency (IGHD); and a group of 154 controls. Heterozygous mutations were identified in all three groups but disproportionately in the individuals with short stature, both with (odds ratio 25.2; 95% CI, 5.1-132.2) and without (odds ratio 3.6; 95% CI, 1.0-12.9) IGHD. Twenty-four novel GH1 gene lesions were found. Thirteen novel missense mutations were characterized by assaying the signal transduction activity of in vitro expressed variants; six (T27I, K41R, N47D, S71F, S108R, and T175A) exhibited a reduced ability to activate the JAK/STAT pathway. Molecular modeling suggested that both K41R and T175A might compromise GH receptor binding. Seven GH variants (R16C, K41R, S71F, E74K, Q91L, S108C, and a functional polymorphism, V110I) manifested reduced secretion in rat pituitary cells after allowance had been made for the level of expression attributable to the associated GH1 proximal promoter haplotype. A further leader peptide variant (L-11P) was not secreted. Eleven novel mutations in the GH1 gene promoter were assessed by reporter gene assay but only two, including a GH2 gene-templated gene conversion, were found to be associated with a significantly reduced level of expression. Finally, a novel intron 2 acceptor splice-site mutation, detected in a family with autosomal dominant type II IGHD, was shown to lead to the skipping of exon 3 from the GH1 transcript. A total of 15 novel GH1 gene mutations were thus considered to be of probable phenotypic significance. Such lesions are more prevalent than previously recognized and although most may be insufficient on their own to account for the observed clinical phenotype, they are nevertheless likely to play a contributory role in the etiology of short stature. PMID- 12655558 TI - Mitochondrial DNA mutation correlates with stage progression and prognosis in non small cell lung cancer. PMID- 12655559 TI - Gene structure of human carbamylphosphate synthetase 1 and novel mutations in patients with neonatal onset. AB - Carbamylphosphate synthetase 1 (E.C. 6.3.4.16) deficiency is a rare autosomal recessive disorder of the urea cycle that can result in severe neonatal hyperammonemia. Since the genomic structure of the CPS1 gene was not yet elucidated, mutation detection was performed by analysis of transcripts in the past. Here, we present the entire DNA sequence of the human CPS1 gene including all exon-intron boundaries. Moreover, mutation analysis was performed in six patients leading to the detection of 9 novel mutations including the missense mutations c.2528T>C and c.2623A>G, the nonsense mutations c.712C>T and c.2115ins35bp, the splice site mutations c.1263+5G>C, c.3558+1G>C and c.4101+2T>C, and a small deletion c.3036_3038delGGT. The mutations c.2528T>C and c.2623A>G were identified on a double mutated allele. New data on the genomic structure of the CPS1 gene provided in this study are useful to characterize the heterogenous molecular basis of the disease in patients deficient for carbamylphosphate 1 deficiency. PMID- 12655560 TI - Germline mutations in the BRCA1 and BRCA2 genes in Turkish breast/ovarian cancer patients. AB - In this study we genotyped Turkish breast/ovarian cancer patients for BRCA1/BRCA2 mutations: protein truncation test (PTT) for exon 11 BRCA1 of and, multiplex PCR and denaturing gradient gel electrophoresis (DGGE) for BRCA2, complemented by DNA sequencing. In addition, a modified restriction assay was used for analysis of the predominant Jewish mutations: 185delAG, 5382InsC, Tyr978X (BRCA1) and 6174delT (BRCA2). Eighty three breast/ovarian cancer patients were screened: twenty three had a positive family history of breast/ovarian cancer, ten were males with breast cancer at any age, in eighteen the disease was diagnosed under 40 years of age, one patient had ovarian cancer in addition to breast cancer and one patient had ovarian cancer. All the rest (n=30) were considered sporadic breast cancer cases. Overall, 3 pathogenic mutations (3/53-5.7%) were detected, all in high risk individuals (3/23-13%): a novel (2990insA) and a previously described mutation (R1203X) in BRCA1, and a novel mutation (9255delT) in BRCA2. In addition, three missense mutations [two novel (T42S, N2742S) and a previously published one (S384F)] and two neutral polymorphisms (P9P, P2532P) were detected in BRCA2. Notably none of the male breast cancer patients harbored any mutation, and none of the tested individuals carried any of the Jewish mutations. Our findings suggest that there are no predominant mutations within exon 11 of the BRCA1 and in BRCA2 gene in Turkish high risk families. PMID- 12655561 TI - A double missense variation of the BUB1 gene and a defective mitotic spindle checkpoint in the pancreatic cancer cell line Hs766T. AB - To determine sequence variations of the BUB1 and BUB1B genes in pancreatic cancer, the entire coding regions of the BUB1 and BUB1B genes were sequenced in pancreatic cancer cell lines and xenografts. Although only polymorphic alterations were found in the BUB1B gene, the aneuploid pancreatic cell line Hs766T had two novel missense variants (p.[Y259C;H265N]) in the BUB1 gene. These mutations were on the same allele, accompanied by a wild-type BUB1 allele. This change was not found in other samples, the literature, or 110 additional chromosomes from a reference population. Compared to two cell lines having microsatellite instability (MIN), the TP53 wild-type pancreatic cell line Hs766T had a defective mitotic spindle checkpoint, indicative of a cell line with chromosomal instability (CIN). Evidence that this checkpoint pathway can be abrogated by mutations in the BUB1 gene (Cahill et al., 1998) supports the suggestion the missense mutations of the BUB1 gene in the Hs766T cell line may contribute to its observed mitotic checkpoint defect. PMID- 12655562 TI - Identification of six novel MSH2 and MLH1 germline mutations in HNPCC. AB - Germline mutations in mismatch repair genes are responsible for hereditary nonpolyposis colorectal cancer (HNPCC), the most common hereditary cancer susceptibility syndrome. We report six novel germline mutations, three in MSH2 and three in MLH1. All but one mutation have been found in families fulfilling the criteria of the Bethesda guidelines; two of them additionally fulfilled the Amsterdam criteria. We identified two nonsense mutations in MSH2 (c.1764T>G [p.Y588X], c.2579C>A [p.S860X]), one duplication of four nucleotides causing premature stop codon (MLH1: c.821_824dupAAGC [p.A275fsX307]), one splice site mutation resulting in skipping of exon 8 from the MLH1 transcript (c.677+3A>G), one duplication of 18 nucleotides leading to duplication of six amino acids in the mismatch-binding domain of MSH2 (c.4_21dup [p.A2_E7dup) and one missense mutation in the PMS2 interaction domain of MLH1 (c.1756G>C [p.A586P]). The three latter mutations were not found in 73, 90 and 94 healthy control individuals, respectively. The corresponding tumors from all patients showed a high level of microsatellite instability (MSI-H). Immunohistochemistry (IH) revealed complete loss of expression of the affected protein in the tumor cells from the patients with the nonsense, splice-site and missense mutation. The tumor from the patient with the c.821_824dupAAGC mutation showed a reduced, rather than lost, expression of the MLH1-protein. PMID- 12655563 TI - Identification of seven novel mutations in the GAN gene. AB - Giant axonal neuropathy (GAN) is a severe early onset neurodegenerative disorder affecting both the peripheral nerves and the central nervous system. The diagnosis is based on the presence of characteristic giant axons on nerve biopsy. In GAN, the integrity of the intermediate filament network is altered. Indeed, abnormal accumulation of the intermediate filaments has been reported in different cell types, including in the swollen axons, which are filled with neurofilaments. We identified the defective protein, gigaxonin, of unknown function, and reported fourteen distinct mutations in twelve families of various origins. Two additional mutations have been recently reported. In the present study, we analysed the GAN gene in 6 families, and identified seven novel mutations: three nonsense and two missense mutations and two deletions. In addition, the molecular result for an already reported family was re-evaluated. In this family, the R269Q "polymorphism" is in fact the pathogenic mutation. PMID- 12655565 TI - Mutation analysis of the entire keratin 5 and 14 genes in patients with epidermolysis bullosa simplex and identification of novel mutations. AB - Epidermolysis bullosa simplex is a group of blistering skin disorders caused by defects in one of the keratin genes, KRT5 and KRT14. Previously reported KRT5 and KRT14 mutations are clustered in several hotspots, namely the rod ends of the 1A and 2B domains and in the non-helical linker region L12. Therefore, genomic KRT5 and KRT14 mutation analysis was initially limited to these hotspots. In this study we describe the screening of nine EBS patients for mutations in the hotspots. In two patients, with the Koebner and the Weber-Cockayne subtypes of epidermolysis bullosa simplex respectively, we could, however, not identify any mutation in one of the hotspot domains of KRT5 and KRT14. Therefore, it appeared to be necessary to screen the entire genes for mutations. For KRT5, a complete genomic mutation detection system was previously described. We now developed a complete genomic mutation detection system for KRT14. For the amplification of the KRT14 genes, we make use of restriction sites to exempt the keratin 14 pseudogene sequence from polymerase chain reaction amplification. Using the complete genomic mutation detection system for both KRT5 and KRT14, we identified four novel KRT5 mutations (IVS1-1G>C, K404E, A438D, E475K), two of which are outside the KRT5 hotspot domains, and three novel KRT14 mutations (IVS4+1G>A, L408M, L130P). PMID- 12655564 TI - Genetic analysis of familial colorectal cancer in Israeli Arabs. AB - Colorectal cancers (CRC) among Israeli Arabs differ from those diagnosed in Jewish Israeli individuals in two manners: an earlier age of occurrence and a low frequency. These differences are unaccounted for and thus prompted us to perform genetic analysis in Israeli Arab CRC patients. Analysis included the major Hereditary non-polyposis colorectal cancer (HNPCC) genes and the APC I1307K mutation (MIM# 175100.0029). Twenty-five patients and 25 relatives from 24 unrelated families were clinically classified according to personal and familial cancer history. If MSI (microsatellite instability) was displayed in tumor tissue, patients underwent mutation analysis of the MSH2 and MLH1 genes using DGGE (denaturing gradient gel electrophoresis) and sequencing. MSI was detected in 9/21 of the tumors tested (43%). Two novel missense mutations were diagnosed among 11 fully analyzed patients: a change of A to G at position 380 in MSH2 (N127S), and a D601G mutation in MLH1. The I1307K mutation was detected in 8 families (8/24, 33.3%). This is the first report of genetic analysis in familial CRC associated genes among Israeli Arabs. We suggest that the I1307K mutation may contribute to CRC in Israeli Arabs and that inactivating mutations of MSH2 and MLH1 may not be a major cause for early onset CRC. PMID- 12655566 TI - Genetic analysis of variegate porphyria (VP) in Italy: identification of six novel mutations in the protoporphyrinogen oxidase (PPOX) gene. AB - Variegate Porphyria (VP) is one of the acute hepatic porphyrias, and is clinically characterised by skin lesions and acute neuropsychiatric/visceral attacks that occur separately or together. The disorder is caused by a partial deficiency of protoporphyrinogen oxidase, the penultimate enzyme in the heme biosynthetic pathway, and a number of mutations have been described for the corresponding gene (PPOX). Here we report a genetic analysis of VP in Italy, and the identification of six novel and three previously characterised mutations from nine affected individuals and families. Among those newly identified, two mutations were small deletions (c.418_419delAA; c.759delA), leading to the formation of premature stop codons, two were splicing defects (IVS10+2T>G; IVS12+1G>C), one was a nonsense (c.384G>A=p.W128X) and one a missense mutation (c.848T>A=I283N). This is the first study of the molecular genetics of Variegate Porphyria in patients of Italian origin, and the finding of six novel mutations out of nine identified confirms the genetic heterogeneity observed for this disorder. PMID- 12655567 TI - Mutational analysis of BRCA2 in Spanish breast cancer patients from Castilla Leon: identification of four novel truncating mutations. AB - Mutations in BRCA1 and BRCA2 account for approximately 5% of the overall familial risk of breast cancer. We have carried out a mutational analysis of the entire coding sequence of the BRCA2 gene in 150 breast cancer patients from Castilla Leon by two different methods: Protein Truncation Test (PTT) and Conformation Sensitive Gel Electrophoresis (CSGE). We have identified 10 distinct truncating mutations of the BRCA2 protein in 17 unrelated patients. Four mutations had not been previously described in any other population: two nonsense, Q2354X and K3083X, and two frameshift deletions, 6126delT and 5374delTATG. Moreover, three further mutations, 1538delAAGA, E1308X and S2219X, had not been reported in Spanish patients until now. Five mutations were recurrent: 3036delACAA, 1538delAAGA, 9538delAA and the novel mutations, 6126delT and 5374delTATG. The most prevalent mutation was 3036delACAA found in four unrelated patients. PMID- 12655568 TI - Novel MLH1 and MSH2 germline mutations in the first HNPCC families identified in Slovakia. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is a dominantly-inherited cancer predisposition syndrome, in which the susceptibility to cancer of the colon, endometrium and ovary is linked to germline mutations in DNA mismatch repair (MMR) genes. We have recently initiated a cancer prevention program in suspected HNPCC families in the Slovak Republic. The first ten families fulfilling Amsterdam criteria or Bethesda guidelines were screened for germline mutations in MLH1 and MSH2, two MMR genes most frequently mutated in HNPCC families. Six mutations were identified, five of which have not been reported previously. Two of the three new mutations in MLH1 (c.380+2T>A; c.307-2A>C) were absent from 100 chromosomes of healthy controls and probably cause a splicing defect, while the third was a 1 bp deletion (c.1261delA). In the MSH2 gene, one new nonsense (c.1030C>T [p.Q344X]) and one missense (c.524T>C [p.L175P]) mutation were identified. This latter variant was not found in 104 alleles of healthy control individuals. Moreover, a previously-reported pathogenic mutation (c.677G>T [p.R226L]) was found in one kindred. The clinical data and the genotypic and phenotypic evaluation of the tumors indicate that all the new alterations are pathogenic HNPCC mutations. PMID- 12655569 TI - Mutational spectrum of the iduronate 2 sulfatase gene in 25 unrelated Korean Hunter syndrome patients: identification of 13 novel mutations. AB - Hunter syndrome (Mucopolysaccharidosis type II, MPS2) is an X-linked recessively inherited disease caused by a deficiency of iduronate 2 sulfatase (IDS). In this study, we investigated mutations of the IDS gene in 25 Korean Hunter syndrome patients. We identified 20 mutations, of which 13 mutations are novel; 6 small deletions (69_88delCCTCGGATCCGAAACGCAGG, 121-123delCTC, 500delA, 877_878delCA, 787delG, 1042_1049delTACAGCAA), 2 insertions (21_22insG, 683_684insC), 2 terminations (529G>T, 637A>T), and 3 missense mutations (353C>A, 779T>C, 899G>T). Moreover, using TaqI or HindIII RFLPs, we found three gene deletions. When the 20 mutations were depicted in a 3-dimensional model of IDS protein, most of the mutations were found to be at structurally critical points that could interfere with refolding of the protein, although they were located in peripheral areas. We hope that these findings will further the understanding of the underlying mechanisms associated with the disease. PMID- 12655570 TI - Six novel ATM mutations in Italian patients with classical ataxia-telangiectasia. AB - Mutations in the ATM gene are responsible for the autosomal recessive syndrome Ataxia Telangiectasia (AT). In a group of 26 classical AT Italian patients studied by protein truncation test (PTT), we identified six new mutations, never reported so far. Mutations -spread over the entire ATM coding sequence with not clear "hot-spot"- are four frameshifts (2192_2193insA, 3110delC, 7150delA, 8368delA), one splice site alteration (8850G>T, causing exon 63 skipping) and one nonsense change (6913C>T, Q2305X). The identification of ATM gene mutations is important for understanding the molecular basis of the disease, and is essential for diagnosis and genetic counseling. PMID- 12655571 TI - Identification of novel SNPs in the interleukin 6 receptor gene (IL6R). AB - The human interleukin-6 receptor (IL6R) is responsible for signal transduction of IL6 that might have associations with immune diseases and various infectious diseases. We have sequenced all 10 exons including the putative splicing site to identify single nucleotide polymorphisms in IL6R. Seven novel single nucleotide polymorphisms (SNPs) were identified; one SNP in the promoter region (-183G>A), one synonymous SNP in exon 2 (24013G>A: Ala31Ala), one non-synonymous SNP in exon 9 (48892A>C: Asp358Ala) and four in introns (29753A>C, 42700T>C, 48869T>A and 59818C>T). The frequencies of each SNP in the Korean population (n=300) were 0.48 (-183 G>A), 0.13 (24013 G>A), 0.41 (29753A>C), 0.41 (42700T>C), 0.1 (48869T>A), 0.42 (48892A>C), and 0.07 (59818C>T), respectively. Haplotypes and their frequencies were estimated by the EM algorithm. Linkage disequilibrium coefficients (/D'/) between each SNP pair were also calculated. The information on SNPs and haplotypes in IL6R would be useful for genetic studies of this gene. PMID- 12655572 TI - Bruton tyrosine kinase gene mutations in Argentina. AB - The block in differentiation from pro-B to pre-B cells results in a selective defect in the humoral immune response characteristic of human X-linked agammaglobulinemia (XLA). Mutations of Bruton tyrosine kinase (BTK) gene have been identified as the cause of XLA. Mutation detection is the most reliable method for making a definitive diagnosis, except when clinical and laboratory findings are distinctive and coupled with history of X-linked inheritance. To provide a definitive diagnosis to 40 families incorporated in the Argentinian Primary Immunodeficiencies Registry we analysed the BTK gene by SSCP analysis as screening method for XLA, followed by direct sequencing. The molecular defect was localized in 45 patients from 34 unrelated families. From the 34 independent mutations identified, 16 were previously undescribed, 31 were unique mutations, 22 were exonic single nucleotide changes (16 missense and 6 nonsense) and four intronic mutations. Because five families had clinical, immunological and inheritance data sufficient for a definitive diagnosis, our study allowed 37 patients from 29 families previously categorized probable/ possible XLA, have now definitive diagnosis leading to appropriate genetic counseling. PMID- 12655573 TI - Mutations in the human homologue of Drosophila patched in Japanese nevoid basal cell carcinoma syndrome patients. AB - Mutations in the human homologue of Drosophila patched (PTCH) have been identified in patients with nevoid basal cell carcinoma syndrome (NBCCS; also called Gorlin syndrome) as well as sporadic basal cell carcinomas and medulloblastomas. However, using PCR-SSCP analysis, mutations in PTCH have been found in only a fraction (about one third to a half) of NBCCS patients. In this study, we determined the whole genomic organizations of the PTCHgene and developed a new set of more accurate primers for the analysis of mutations in PTCH. Using these primers, we examined 8 Japanese NBCCS patients for mutations in all PTCH exons by direct sequencing of the PCR products. As a result, we identified 5 novel PTCH mutations in 6 out of 8 patients including 2 sisters as well as 5 polymorphisms, two of them, 1704G>C and 2928G>C were novel. Four of these mutations, 900delC, 1247insT, 1999delC and 933+5G>T, cause protein truncation due to the insertion or deletion of a single nucleotide or aberrant splicing. The remaining mutation, 1514G>A was a missense alteration (G509D). Interestingly, the amino acid substitution, G509V, has been reported previously in an NBCCS patient, suggesting an important role of this amino acid residue in the function of PTCH protein. The difference in the detection rate of PTCH mutations among NBCCS between previous reports and ours is due to the difference either in ethnicity or in the detection methods. PMID- 12655574 TI - Haplotype analysis of the BRCA2 9254delATCAT recurrent mutation in breast/ovarian cancer families from Spain. AB - A frame-shift 9254del5 mutation was independently identified in 12 families, eleven of them with Spanish ancestors, in a BRCA2 screening performed in 841 breast and/or ovarian cancer families and in 339 women with breast cancer diagnosed before the age of 40 at different centers in France and Spain. We sought to analyze in detail the haplotype and founder effects of the 9254del5 and to estimate the time of origin of the mutation. Eight polymorphic microsatellite markers and two BRCA2 polymorphisms were used for the haplotype analyses. The markers were located flanking the BRCA2 gene spanning a region of 6.1 cM. Our results suggest that these families shared a common ancestry with BRCA2 9254del5, which is a founder mutation originating in the Northeast Spanish, with an estimated age of 92 (95% CI 56-141) generations. PMID- 12655575 TI - Pathogenic mutations of the lipoprotein lipase gene in Chinese patients with hypertriglyceridemic type 2 diabetes. AB - Elevated plasma triglyceride and nonesterified fatty acid concentrations may cause insulin resistance. Lipoprotein lipase (LPL) is a rate-determining enzyme in lipid metabolism. To investigate the role of the LPL gene in Chinese patients with hypertriglyceridemic type 2 diabetes, 277 patients with type 2 diabetes and 241 healthy control subjects were recruited and screened for sequence changes in the LPL gene by PCR, SSCP, restriction analysis and direct DNA sequencing. Ten mutations were identified: four missense mutations, Ala71Thr, Val181Ile, Gly188Glu and Glu242Lys; one nonsense mutation Ser447Ter; and five silent mutations. Ser447Ter was found in both patients and controls with no significant difference in frequency. The four missense mutations were located in the highly conserved exon 3, 5, and 6 regions and in highly conserved amino acid sites. They led to reduced LPL mass and enzyme activities in both post-heparin plasma and in vitro expression. The modeled structures displayed major differences between the mutant and wildtype molecules. These results indicated that the four missense mutations lead to LPL deficiency and subsequent hypertriglyceridemia. Based on our study and published data, a putative pathogenic pathway was suggested: LPL enzyme deficiency causes elevated plasma triglyceride level and subsequent insulin resistance; both increased free fatty acids and insulin resistance promote gluconeogenesis and hyperglycaemia, a vicious circle leading to type 2 diabetes. PMID- 12655576 TI - Mutation analysis in 16 patients with mtDNA depletion. AB - Sixteen unrelated Southern European patients with the mitochondrial depletion syndrome (MDS) were analyzed for mutations in the TK2 and DGUOK genes. Three novel mutations were identified in TK2 (R183G, R254X, and 142insG). When we analyzed additional genes involved in the dNTPs pool, such as SLC25A19 (DNC) and NT5M (d-NT2), we did not detect mutations. The current study suggest that scanning the TK2, DGUOK, SLC25A19, and NT5M genes is likely to help about 10% of MDS families in terms of genetic counseling. Also, our findings indicate that genotype-phenotype correlations are not straightforward in MDS. PMID- 12655577 TI - Guidelines for using quantitative magnetization transfer magnetic resonance imaging for monitoring treatment of multiple sclerosis. AB - Quantitative evaluation of brain magnetic resonance imaging (MRI) scans is now an accepted part of the trial of new putative treatments for multiple sclerosis (MS). However, conventional MRI is not pathologically specific, and it does not reveal the details of the pathological processes that underlie the progression of the disease. Magnetization transfer (MT) imaging is a relatively new quantitative technique that appears to offer some pathological specificity, and can be used to monitor the changes over time in both individual lesions and the central nervous system as a whole. This paper considers the case for incorporating MT imaging into new clinical trials, so that the utility of MT for monitoring the modification of MS progression by treatment can be assessed. Specific guidelines for implementing MT imaging as part of a large multicenter clinical trial are given, and practical considerations when planning such a trial are detailed. It is anticipated that MT imaging will be incorporated into many new trials in the near future. PMID- 12655578 TI - MRI tissue characterization of experimental cerebral ischemia in rat. AB - PURPOSE: To extend the ISODATA image segmentation method to characterize tissue damage in stroke, by generating an MRI score for each tissue that corresponds to its histological damage. MATERIALS AND METHODS: After preprocessing and segmentation (using ISODATA clustering), the proposed method scores tissue regions between 1 and 100. Score 1 is assigned to normal brain matter (white or gray matter), and score 100 to cerebrospinal fluid (CSF). Lesion zones are assigned a score based on their relative levels of similarities to normal brain matter and CSF. To evaluate the method, 15 rats were imaged by a 7T MRI system at one of three time points (acute, subacute, chronic) after MCA occlusion. Then they were killed and their brains were sliced and prepared for histological studies. MRI of two or three slices of each rat brain (using two DWI (b = 400, b = 800), one PDWI, one T2WI, and one T1WI) was performed, and an MRI score between 1 and 100 was determined for each region. Segmented regions were mapped onto the histology images and scored on a scale of 1-10 by an experienced pathologist. The MRI scores were validated by comparison with histology scores. To this end, correlation coefficients between the two scores (MRI and histology) were determined. RESULTS: Experimental results showed excellent correlations between MRI and histology scores at different time points. Depending on the reference tissue (gray matter or white matter) used in the standardization, the correlation coefficients ranged from 0.73 (P < 0.0001) to 0.78 (P < 0.0001) using the entire dataset, including acute, subacute, and chronic time points. This suggests that the proposed multiparametric approach accurately identified and characterized ischemic tissue in a rat model of cerebral ischemia at different stages of stroke evolution. CONCLUSION: The proposed approach scores tissue regions and characterizes them using unsupervised clustering and multiparametric image analysis techniques. The method can be used for a variety of applications in the field of computer-aided diagnosis and treatment, including evaluation of response to treatment. For example, volume changes for different zones of the lesion over time (e.g., tissue recovery) can be evaluated. PMID- 12655579 TI - In vivo accuracy of multisequence MR imaging for identifying unstable fibrous caps in advanced human carotid plaques. AB - PURPOSE: To evaluate the in vivo accuracy of a multisequence MRI technique for prospectively identifying one feature of the vulnerable plaque-an unstable fibrous cap-in human carotid atherosclerosis. MATERIALS AND METHODS: The carotid arteries of 18 endarterectomy patients were preoperatively imaged in a 1.5 T scanner using a multisequence protocol that generated four contrast weightings (3D time of flight (ToF), T1, proton density (PD), and T2) at each slice location. With the use of previously published MR criteria, the images of the vessel wall were first examined for evidence of an unstable fibrous cap. The imaging findings were then correlated with the histology from the surgical specimens. RESULTS: A blinded review of the MR findings with the histologic state of the fibrous cap revealed that 1). assessing the preoperative appearance of the fibrous cap has a high test sensitivity (0.81) and specificity (0.90) for identifying an unstable cap in vivo; and 2). the availability of different contrast weightings facilitated image interpretation when intimal calcifications or flow artifacts obscured the lumen surface. CONCLUSION: Multisequence MRI can accurately characterize the in vivo state of the fibrous cap. This finding supports the use of these noninvasive techniques to prospectively identify vulnerable plaques. PMID- 12655580 TI - Flow-independent magnetic resonance venography of the calf. AB - PURPOSE: To show that flow-independent venography that exploit the intrinsic MR properties of blood to isolate vessels from surrounding structures can be used for depiction of peripheral veins and for detection of deep venous thrombosis (DVT). MATERIALS AND METHODS: Sequence and parameters were first determined on a theoretical basis. The sequence was then optimized in volunteers (N = 4) for the depiction of the peripheral venous vessels. Qualitative evaluation of the normal venous anatomy was performed in five volunteers. The feasibility of diagnosing DVT of the calf with this method was evaluated with preliminary clinical studies. RESULTS: Excellent depiction of the venous anatomy was achieved in all volunteers with the optimized technique. Very small venous structures, such as superficial, muscular, and perforator veins, were clearly depicted because of the high spatial and contrast resolution capacities of the sequences. In all six patients, DVT findings diagnosed by duplex sonography were also seen on MR venography. CONCLUSION: Venous anatomy mapping and detection of lower-limb DVT appear feasible using flow-independent MR venograms. PMID- 12655581 TI - Dynamic MRI of breast hardness following radiation treatment. AB - PURPOSE: To evaluate functional microvascular characteristics of breast induration several years after radiation treatment using dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) techniques. MATERIALS AND METHODS: Fifteen women with moderate or marked breast induration after surgery and radiotherapy for breast cancer (2-15 years) were examined. Images of the irradiated breast (boost and nonboost sites) on short tau inversion recovery (STIR) and DCE-MRI sequences were subjectively evaluated for edema and the presence of enhancement and compared to the contralateral normal breast. Quantitative enhancement parameters-percent enhancing pixels, transfer constant (K(trans)), rate constant (k(ep)), leakage space (v(e)), and maximum contrast medium accumulation (MCMA)-were also compared. RESULTS: No tumor recurrence was seen. Fat necrosis was seen in 2/15 cases. Increased parenchymal edema at the electron boost site was seen in 12/14 patients. Greater enhancement in the irradiated breast was seen in 11/14 evaluable patients. Kinetic parameter estimates including K(trans) were similar except for percent enhancing pixels, which was greater in the irradiated breast at both boost and nonboost sites (P = 0.03 and 0.04, respectively). v(e) and MCMA estimates were greater in breasts with marked induration compared to moderate grades (P = 0.002 and 0.01, respectively). CONCLUSION: Parenchymal edema may be an important contributor to palpable induration several years after breast radiotherapy. Increased fluid content may be related to increased numbers of perfused microvessels and/or impaired lymphatic drainage. PMID- 12655582 TI - Magnetic resonance microscopy quantifies the disease progression in Marfan syndrome mice. AB - PURPOSE: To use noninvasive magnetic resonance microscopy (MRM) to examine aneurysmal disease in the mouse model of Marfan syndrome (MFS). MATERIALS AND METHODS: A total of eight wild-type (WT) and MFS mice were imaged using MRM; four of them at three different time points over an 8-week period and the remaining animals were imaged at one time point. The maximal cross-sectional area of the aorta was measured by manual tracing and by automated means from combined cardiac and respiratory-gated bright-blood images. Relationships between aortic size and age and the differences between WT and mutant mice aortic size were established. RESULTS: Maximal cross-sectional aortic areas differed significantly (P < 0.05) between WT and mutant mice for all time points, with MFS mice having larger aortic size. There was a positive correlation between aortic size and age in MFS mice (r = 0.80) with a significant increase from the 14th to the 22nd week (P < 0.05). CONCLUSION: MRM detected the differences in aortic size between WT and mutant mice over time, demonstrating a potential for noninvasive technique for the assessment of potential therapies in MFS mice. PMID- 12655583 TI - Investigation of apparent diffusion constant as an indicator of early degenerative disease in articular cartilage. AB - PURPOSE: To investigate the apparent diffusion constant (ADC) as a prospective magnetic resonance imaging (MRI) marker of early degeneration in articular cartilage. MATERIALS AND METHODS: Early degenerative changes were studied using in vitro MRI on cartilage-bone specimens excised from human femoral condyles. The loss of proteoglycans developed in vivo due to a degenerative process was compared with a gadolinium diethylenetriamine pentaacetate anion (Gd-DTPA(2-)) enhanced decrease of T(1) relaxation times, and with an increase of ADCs and T(2) relaxation times. RESULTS: Contrast enhanced T(1) values decreased and the diffusion constants increased in cartilage regions with depleted proteoglycans. The relative changes in diffusion constants were smaller than those of Gd-DTPA(2 ) enhanced T(1), and in some proteoglycan-depleted regions no changes in the diffusion constants were detected. T(2) relaxation times showed considerable spatial variability that did not correlate with proteoglycan concentration. CONCLUSION: In contrast to Gd-DTPA(2-) enhanced T(1), which reflects changes in chemical composition, diffusion constants may reflect structural degradation of the cartilage matrix. PMID- 12655585 TI - Challenges in dynamic contrast-enhanced MRI imaging of cervical lymph nodes to detect metastatic disease. AB - PURPOSE: To identify and overcome challenges in using dynamic contrast-enhanced magnetic resonance imaging (MRI) to distinguish tumor from nontumor in the cervical lymph nodes of patients with squamous cell carcinoma of the head and neck. MATERIALS AND METHODS: Signal-to-noise ratio (S/N), motion, node heterogeneity, and tissue normalizations were examined. Twenty-one patients with squamous cell carcinoma of the head and neck were scanned before a neck dissection (two-dimensional fast spoiled gradient-echo: 10 locations/13 seconds). Peak time, peak enhancement, maximum upslope, and washout slope were measured in pathologically confirmed tumor and nontumor nodes and in the submandibular gland and the sternocleidomastoid muscle. RESULTS: Surface coil arrays provided high coverage and high S/N. Motion averaged 1.1 pixels and was corrected. Large tumor nodes were heterogeneous in their contrast enhancement, while the nontumor nodes were homogeneous. The contrast enhancement parameters were significantly different for all regions except for the submandibular gland compared to the nontumor nodes. CONCLUSION: Challenges of dynamic imaging of cervical lymph nodes were overcome and significant differences were found between the tumor and nontumor nodes, indicating that dynamic imaging is feasible and may aid this patient population. PMID- 12655584 TI - Tumor vascular architecture and function evaluated by non-invasive susceptibility MRI methods and immunohistochemistry. AB - PURPOSE: To investigate the physiological origins responsible for the varying blood oxygenation level dependent (BOLD) magnetic resonance imaging (MRI) responses to carbogen (95% O(2)/5% CO(2)) breathing observed with different tumor types. MATERIALS AND METHODS: Susceptibility contrast-enhanced MRI using the exogenous blood pool contrast agent NC100150 to determine blood volume and vessel size, and immunohistochemical-derived morphometric parameters, were determined in GH3 prolactinomas and RIF-1 fibrosarcomas, both grown in mice, which exhibited very different BOLD responses to carbogen. RESULTS: Administration of NC100150 increased the R(2)* and R(2) rates of both tumor types, and indicated a significant four-fold larger blood volume in the GH3 tumor. The ratio deltaR(2)*/deltaR(2) showed that the capillaries in the GH3 were two-fold larger than those in the RIF-1, in agreement with morphometric analysis. Carbogen breathing induced a significant 25% decrease in R(2)* in the GH3 prolactinoma, whereas the response in the RIF-1 fibrosarcoma was negligible. CONCLUSION: Low blood volume and small vessel size (and hence reduced hematocrit) are two reasons for the lack of R(2)* change in the RIF-1 with carbogen breathing. BOLD MRI is sensitive to erythrocyte-perfused vessels, whereas exogenous contrast agents interrogate the total perfused vascular volume. BOLD MRI, coupled with a carbogen challenge, provides information on functional, hemodynamic tumor vasculature. PMID- 12655586 TI - Assessment of reliability in functional imaging studies. AB - PURPOSE: To investigate the reliability of functional magnetic resonance imaging (fMRI), an approach for mapping and quantifying reliably activated voxels was developed. MATERIALS AND METHODS: First, a SPM99 analysis was performed, and the resulting statistical maps were taken as the basis for subsequent analyses of reliability. Several approaches were demonstrated using 1). a voxel-wise intraclass correlation coefficient (ICC); 2). an analysis of scatter plots, calculating the correlation of contrast t-values for pairs of activation maps; and 3). the ratio of overlapping volumes as suggested in the literature. The methods were applied to an fMRI study in which subjects were asked to vary their attentional effort during watching a flickering checkerboard pattern with varying letters in the center. The subjects had to ignore or attend to the presentation, or they had to detect a target letter within the checkerboard. RESULTS: The imaging data showed good reliability in terms of ICC for regions of visual processing, as well as for frontal areas, especially in the letter detection task. Furthermore, the size of reliable clusters depended on the presumed attentional effort of the subjects. CONCLUSION: Application of the method demonstrated that the activation due to visual stimulation could also be detected very consistently during a no-attend condition, but the reliability of the activations were best during the attended tasks. PMID- 12655587 TI - Calibrated magnetic resonance hydrometry: an in vitro study. AB - PURPOSE: To demonstrate a quantitative approach to measuring fluid volumes with standard single shot RARE sequences. MATERIALS AND METHODS: In phantom experiments, magnetic resonance hydrometry (MRH), in combination with various calibration phantoms (5 mL up to 500 mL) as internal standards, was used to quantify fluid volumes. In total, 16 volume phantoms were investigated with six different calibration phantoms. In addition, noise correction was implemented to correct the quantification results and to avoid the influence of random noise in the image. RESULTS: All MR measurements show significant correlations of up to r = 0.99 (P <.05) with the real applied volume in the investigated phantoms. However, measurements of large volumes were more precise with large calibration phantoms. Noise reduction did not change the correlation between measured and real applied volumes, but did reduce the error of the measured volumes. Calibrated magnetic resonance hydrometry (cMRH) is able to quantify volumes of fluid fast and noninvasively. The volumes of the used calibration phantoms have to be at least in the order of magnitude of the volumes that are to be measured. CONCLUSION: In vitro, cMRH using a single-shot rapid acquisition with refocused echoes (ssRARE) sequence and calibration phantoms is a fast and accurate method of quantifying steady amounts of fluid. PMID- 12655588 TI - Multislice double inversion-recovery black-blood imaging with simultaneous slice reinversion. AB - PURPOSE: To develop a technique for time-efficient multislice double inversion recovery (DIR) black-blood imaging and to test its applicability and limitations for high-resolution imaging of carotid arteries. MATERIALS AND METHODS: A multislice DIR pulse sequence with fast spin-echo (FSE) readout was implemented on a 1.5 T magnetic resonance (MR) scanner. The principle of the method is that a slice-selective inversion in a DIR preparation reinverts an entire slice pack, and all slices are imaged within repetition time (TR). The number of slices acquired per TR (N) controls the inversion time (TI) to execute the readout for each slice at the zero-crossing point of blood. Multislice DIR images (TR/TE = 2500/9 msec) of carotid arteries were obtained with variable N = 2-8 from four subjects. The method was compared with the standard single-slice DIR and inflow saturation techniques. RESULTS: Multislice DIR with N = 2-6 provided similar flow suppression in carotid arteries as single-slice DIR. At all N = 1-8, blood suppression by DIR was significantly better than by inflow saturation. An additional limitation of multislice DIR was saturation of the signal from stationary tissues that worsened visualization of the vessel wall at N >or= 6. CONCLUSION: Multislice DIR provides up to eight-fold improvement of time efficiency relative to single-slice DIR and high-quality blood suppression. PMID- 12655589 TI - MRI depiction of chronic intradural (subdural) hematoma in evolution. AB - The usual chronic subdural hematoma encountered in the elderly is actually thought to be a chronic intradural hematoma, which has been described clinically and anatomically. However, the evolution of how this chronic hematoma occurs remains enigmatic. We report the first magnetic resonance (MR) depiction of an apparent chronic intradural hematoma in evolution over several months in a 61 year-old man after a minimal head injury. The time delay from injury, subsequent focal pachymeningeal T1-gadolinium enhancement, and apparent splitting of the pachymeninges by proven chronic hematoma should serve as a stimulus for further MR investigations of this interesting process. PMID- 12655590 TI - Juxtapapillary diverticulum: findings on MRI. AB - The purpose of our study was to describe the imaging findings of juxtapapillary diverticulum on magnetic resonance imaging (MRI). The MRI and magnetic resonance cholangiopancreatography (MRCP) examinations of 14 patients with juxtapapillary diverticula that were diagnosed on endoscopic retrograde cholangiopancreatography (ERCP) (N = 8) or endoscopy (N = 6) were retrospectively evaluated. T1-weighted spoiled gradient-echo, T2-weighted half Fourier single shot fast spin-echo (HASTE), and T2-weighted True FISP (fast imaging with steady state precession) images and thin-slice MRCP images were obtained on all patients. In five patients, diluted gadolinium DPTA (1/100) was used as an oral contrast. T2 weighted True FISP and HASTE images demonstrated air-fluid levels within all diverticula. Hyperintense oral contrast on T1-weighted spoiled gradient-echo images aided detection of the smaller diverticula. MRCP images obtained in the coronal plane best demonstrated the relationship of the diverticula to the papilla. MRI with the use of HASTE, True FISP, and oral contrast-enhanced T1 weighted sequences was able to depict juxtapapillary diverticula in our series. PMID- 12655591 TI - A manipulator system for 14-gauge large core breast biopsies inside a high-field whole-body MR scanner. AB - PURPOSE: To investigate a robotic manipulator system for MR-guided breast biopsies inside a 1.5 T whole-body magnet. MATERIALS AND METHODS: Fourteen patients with suspicious breast lesions (diameter 18.6 +/- 12 mm) underwent biopsy. Patients with malignant findings underwent surgery afterwards and the histologic findings between biopsy and surgery were correlated. RESULTS: In five patients biopsy findings were histopathologically confirmed following open surgery. One tubular carcinoma was missed; one invasive cancer was underestimated. Seven patients with benign findings are still in the follow-up period. CONCLUSION: The study demonstrates the feasibility to perform breast biopsies inside the magnet of a whole-body MR scanner by using a manipulator system. PMID- 12655592 TI - Time-resolved three-dimensional phase-contrast MRI. AB - PURPOSE: To demonstrate the feasibility of a four-dimensional phase contrast (PC) technique that permits spatial and temporal coverage of an entire three dimensional volume, to quantitatively validate its accuracy against an established time resolved two-dimensional PC technique to explore advantages of the approach with regard to the four-dimensional nature of the data. MATERIALS AND METHODS: Time-resolved, three-dimensional anatomical images were generated simultaneously with registered three-directional velocity vector fields. Improvements compared to prior methods include retrospectively gated and respiratory compensated image acquisition, interleaved flow encoding with freely selectable velocity encoding (venc) along each spatial direction, and flexible trade-off between temporal resolution and total acquisition time. RESULTS: The implementation was validated against established two-dimensional PC techniques using a well-defined phantom, and successfully applied in volunteer and patient examinations. Human studies were performed after contrast administration in order to compensate for loss of in-flow enhancement in the four-dimensional approach. CONCLUSION: Advantages of the four-dimensional approach include the complete spatial and temporal coverage of the cardiovascular region of interest and the ability to obtain high spatial resolution in all three dimensions with higher signal-to-noise ratio compared to two-dimensional methods at the same resolution. In addition, the four-dimensional nature of the data offers a variety of image processing options, such as magnitude and velocity multi-planar reformation, three-directional vector field plots, and velocity profiles mapped onto selected planes of interest. PMID- 12655594 TI - Microglia activation influences dye coupling and Cx43 expression of the astrocytic network. AB - Under inflammatory conditions, activated microglia are capable of producing proinflammatory cytokines that are reported to influence cell-to-cell communication. The present study was performed to evaluate the influence of microglial activation on the coupling efficiency of the astroglial network. Primary astrocyte cultures of newborn rats were cocultured with either 5% (M5) or 30% (M30) microglia. Microglial activation (rounded phagocytotic phenotype) was investigated using the monoclonal anti-ED1 antibody, and immunofluorescence with a polyclonal anti-Cx43 antibody was used to study astroglial Cx43 expression and distribution. Functional coupling of astrocytes was evaluated by monitoring the transfer of microinjected Lucifer yellow into neighboring cells. The data obtained can be summarized as follows: astroglia/M30 cocultures contained significantly fewer resting microglia and significantly more activated microglia than the M5 cocultures; significantly reduced astroglial Cx43 staining was found in M30 cocultures concurrently with a reduced number of dye coupled astrocytes; and the positive correlation of percent activated microglia with reduced astroglial Cx43 expression was highly significant, indicating that the degree of intercellular communication in the astroglial network may be modulated by the activation of microglia under in vitro conditions. PMID- 12655595 TI - Differentiation of radial glia-like cells from embryonic stem cells. AB - Radial glial cells play important roles in neural development. They provide support and guidance for neuronal migration and give rise to neurons and glia. In vitro, neurons, astrocytes, and oligodendrocytes can be generated from neural and embryonic stem cells, but the generation of radial glial cells from these stem cells has not yet been reported. Since the differentiation of radial glial cells is indispensable during brain development, we hypothesize that stem cells also generate radial glial cells during in vitro neural differentiation. To test this hypothesis, we utilized five different clones of mouse embryonic (ES) and embryonal carcinoma (EC) stem cell lines to investigate the differentiation of radial glial cells during in vitro neural differentiation. Here, we demonstrate that radial glia-like cells can be generated from ES/EC cell lines. These ES/EC cell-derived radial glia-like cells are similar in morphology to radial glial cells in vivo, i.e., they are bipolar with an unbranched long process and a short process. They also express several cytoskeletal markers, such as nestin, RC2, and/or GFAP, that are characteristics of radial glial cells in vivo. The processes of these in vitro generated radial glia-like cells are organized into parallel arrays that resemble the radial glial scaffolds in neocortical development. Since radial glia-like cells were observed in all five clones of ES/EC cells tested, we suggest that the differentiation of radial glial cells may be a common pathway during in vitro neural differentiation of ES cells. This novel in vitro model system should facilitate the investigation of regulation of radial glial cell differentiation and its biological function. PMID- 12655596 TI - Microfilament and microtubule organization and dynamics in process extension by central glia-4 oligodendrocytes: evidence for a microtubule organizing center. AB - Microfilaments in freshly adhering CG-4 cells and differentiated CG-4 oligodendrocytes are concentrated at the tips and edges of rapidly forming processes while microtubules are concentrated in new processes and extend from a concentrated spot of alpha-tubulin staining in the cell body to the cell periphery. In motile bipolar CG-4 cells, microfilaments are heavily concentrated at the flattened end of one process and along the rim of processes and the cell body: microtubules are concentrated along main processes and splay out into process tips and the cell body. In differentiated CG-4 oligodendrocytes, microfilaments are concentrated at the many process tips, in filopodia and in fine processes, but are not obvious in main processes where separate bundles of microtubules, which diverge at process branch points, are concentrated. gamma tubulin, involved in microtubule nucleation, is concentrated at a small discrete area in the cell body, indicative of a microtubule organizing center. Polymerization of both actin and tubulin is required for initial process elaboration. Depolymerization of microtubules, but not of microfilaments, causes complete retraction of bipolar CG-4 cell processes. This process retraction does not occur if microfilaments are depolymerized first, indicating that process extension/retraction in motile bipolar CG-4 cells may occur by a balance of motor protein-driven forces as suggested for growth cone motility. Cytoskeleton organization in CG-4 cells is very similar to that reported for oligodendrocytes. CG-4 cells are thus a useful model for investigating the signals and mechanisms regulating oligodendrocyte process dynamics. PMID- 12655597 TI - Slit and glypican-1 mRNAs are coexpressed in the reactive astrocytes of the injured adult brain. AB - The slit family serves as a repellent for growing axons toward correct targets during neural development. A recent report describes slit mRNAs expressed in various brain regions in adult rats. However, their functions in the adult nervous system remain unknown. In the present study, we investigated whether slit mRNAs were expressed in the cryo-injured brain, using in situ hybridization. All slit family members were expressed at the lesion. Slit2 mRNA was the most intensely expressed in the cells surrounding the necrotic tissue. A double labeling study showed that slit2 mRNA was expressed in the glial fibrillary acidic protein (GFAP)-positive reactive astrocytes. In addition, glypican-1, a heparan sulfate proteoglycan that serves as a high-affinity receptor for Slit protein, was coexpressed with slit2 mRNA in the reactive astrocytes. These findings suggested that slit2 might prevent regenerating axons from entering into the lesion in concert with glypican-1. PMID- 12655598 TI - Role of the alpha-chemokine stromal cell-derived factor (SDF-1) in the developing and mature central nervous system. AB - alpha-chemokines, which control the activation and directed migration of leukocytes, participate in the inflammatory processes in host defense response. One of the alpha-chemokines, CXCL12 or stromal cell-derived factor 1 (SDF-1), not only regulates cell growth and migration of hematopoietic stem cells but may also play a central role in brain development as we discuss here. SDF-1 indeed activates the CXCR4 receptor expressed in a variety of neural cells, and this signaling results in diverse biological effects. It enhances migration and proliferation of cerebellar granule cells, chemoattracts microglia, and stimulates cytokine production and glutamate release by astrocytes. Moreover, it elicits postsynaptic currents in Purkinje cells, triggers migration of cortical neuron progenitors, and produces pain by directly exciting nociceptive neurons. By modulating cell signaling and survival during neuroinflammation, SDF-1 may also play a role in the pathogenesis of brain tumors, experimental allergic encephalitis, and the nervous system dysfunction associated with acquired immunodeficiency syndrome. PMID- 12655599 TI - Pharmacological characterization of P2Y receptor subtypes on isolated tiger salamander Muller cells. AB - Muller cells express a variety of neurotransmitter receptors that permit them to "sense" the extracellular environment within the retina. We have used a battery of agonists and antagonists to characterize the purinergic receptor subtypes expressed on isolated tiger salamander Muller cells. Changes in intracellular calcium ion concentration ([Ca(2+)](i)) in Muller cells were measured using the Ca(2+) indicator dye Fura-2 and digital imaging microscopy. ATP, 2-methylthio ATP, 2-methylthio-ADP, ADP, UTP, UDP, deoxyATP, and 3'-O-(4-benzoyl)benzoyl ATP evoked increases in [Ca(2+)](i) in both the presence and absence of extracellular Ca(2+). Therefore, the increases we observed were likely due to intracellular Ca(2+) release mediated by G-protein-coupled P2Y receptor activation, rather than Ca(2+) influx via P2X receptor channels. The P2Y(1) receptor agonists 2 methylthio-ATP, 2-methylthio-ADP, and ADP evoked increases in [Ca(2+)](i) that were inhibited by the P2Y(1) receptor antagonists adenosine 3'-phosphate 5' phosphosulfate and 2'-deoxy-N(6)-methyleneadenosine-3',5'-bisphosphate. Responses to ADP were not completely inhibited by the P2Y(1) receptor antagonists. The residual response to ADP could be mediated by P2Y(13) receptors. UTP evoked an increase in [Ca(2+)](i) that was partially inhibited by suramin, suggesting that Muller cells express P2Y(2) and P2Y(4) receptors. The P2Y(6) receptor agonist UDP, and the P2Y(11) receptor agonists deoxyATP, and 3'-O-(4-benzoyl)benzoyl ATP, evoked increases in [Ca(2+)](i) in Muller cells. We conclude that isolated tiger salamander Muller cells express P2Y(1), P2Y(2), P2Y(6), P2Y(11), and possibly P2Y(4) and P2Y(13) receptors. Therefore, the physiological release of ATP, ADP, UTP, and UDP and/or their accumulation in the retina under pathological conditions could stimulate increases in [Ca(2+)](i) in Muller cells. PMID- 12655601 TI - Temporal progressive antigen expression in radial glia after contusive spinal cord injury in adult rats. AB - In the development of the CNS, radial glial cells are among the first cells derived from neuroepithelial cells. Recent studies have reported that radial glia possess properties of neural stem cells. We analyzed the antigen expression and distribution of radial glia after spinal cord injury (SCI). Sprague-Dawley rats had a laminectomy at Th11-12, and spinal cord contusion was created by compression with 30 g of force for 10 min. In the injury group, rats were examined at 24 h and 1, 4, and 12 weeks after injury. Frozen sections of 20 microm thickness were prepared from regions 5 and 10 mm rostral and caudal to the injury epicenter. Immunohistochemical staining was performed using antibodies to 3CB2 (a specific marker for radial glia), nestin, and glial fibrillary acidic protein (GFAP). At 1 week after injury, radial glia that bound anti-3CB2 MAb had spread throughout the white matter from below the pial surface. From 4 weeks after injury, 3CB2 expression was also observed in the gray matter around the central canal, and was especially strong around the ependymal cells and around blood vessels. In double-immunohistochemical assays for 3CB2 and GFAP or 3CB2 and nestin, coexpression was observed in subpial structures that extended into the white matter as arborizing processes and around blood vessels in the gray matter. The present study demonstrated the emergence of radial glia after SCI in adult mammals. Radial glia derived from subpial astrocytes most likely play an important role in neural repair and regeneration after SCI. PMID- 12655602 TI - Active Src expression is induced after rat peripheral nerve injury. AB - The non-receptor-type Src tyrosine kinases are key components of intracellular signal transduction that are expressed at high levels in the nervous system. To improve understanding of the cascades of molecular events underlying peripheral nerve regeneration, we analyzed active Src expression in the crushed or cut rat sciatic nerves using a monoclonal antibody (clone 28) that recognizes the active form of Src tyrosine kinases, including c-Src and c-Fyn. Western blots showed that active Src expressed in the normal sciatic nerve transiently increased up to threefolds after both types of injury. Immunohistochemistry using clone 28 showed that axonal components are the primary sites of active Src expression in the normal sciatic nerve. Soon after both types of injury, active Src was abundantly expressed in Schwann cells of the segments distal to the injury site. The expression of active Src in the cells decreased with restoration of the axon Schwann cell relationship and eventually became depleted to very low levels after crushing, but was sustained at high levels in the cut model until the end of the experiment. Regenerated axons consistently expressed active Src throughout nerve regeneration and these eventually became the major sites of active Src expression in the crushed nerve. Among the Src tyrosine kinases, active c-Src selectively increased after crushing according to immunoprecipitation and immunoblotting analyses. Due to its potent biological activity, the increased amounts of the active form of Src probably enhance axonal regrowth, the Schwann cell response, and axon-Schwann cell contact for peripheral nerve regeneration. PMID- 12655600 TI - Acute downregulation of Cx43 alters P2Y receptor expression levels in mouse spinal cord astrocytes. AB - Propagation of intercellular calcium waves (ICW) between astrocytes depends on the diffusion of signaling molecules through gap junction channels and diffusion through the extracellular space of neuroactive substances acting on plasmalemmal receptors. The relative contributions of these two pathways vary in different brain regions and under certain pathological conditions. We have previously shown that in wild-type spinal cord astrocytes, ICW are primarily gap junction dependent, but that deletion of the main gap junction protein (Cx43) by homologous recombination results in a switch in mode of ICW propagation to a purinoceptor-dependent mechanism. Such a compensatory mechanism for ICW propagation was related to changes in the pharmacological profile of P2Y receptors, from an adenine-sensitive P2Y(1), in wild-type, to a uridine-sensitive P2U receptor subtype, in Cx43 knockout (KO) astrocytes. Using oligonucleotide antisense to Cx43 mRNA for acute downregulation of connexin43 expression levels, we provide evidence for the molecular nature of such compensatory mechanism. Pharmacological studies and Western blot analysis indicate that there is a reciprocal regulation of P2Y(1) and P2Y(4) expression levels, such that downregulation of Cx43 leads to decreased expression of the adenine-sensitive P2Y(1) receptor and increased expression of the uridine-sensitive P2Y(4) receptor. This change in functional expression of the P2Y receptor subtype population in acutely downregulated Cx43 was paralleled by changes in the mode of ICW propagation, similar to that previously observed for Cx43 KO spinal cord astrocytes. On the basis of these results, we propose that Cx43 regulates both modes of ICW by altering P2Y receptor subtype expression in addition to providing intercellular coupling. PMID- 12655603 TI - Identification of differentially expressed proteins in human glioblastoma cell lines and tumors. AB - An in-frame deletion of 801 bp in exons 2-7 (type III mutation) of the epidermal growth factor receptor (EGFR) is detected at high incidence in primary glioblastoma tumors. A proteomic approach was used to generate differential protein expression maps of fetal human astrocytes (FHA), human glioblastoma cell lines U87MG and U87MG expressing type III EGFR deletion (U87MGdeltaEGFR) that confers high malignancy to tumor cells. Two-dimensional gel electrophoresis followed by in-gel digestion of separated spots and protein identification by LC MS-MS and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) identified 23 proteins expressed at higher levels or exclusively in FHA and 29 proteins expressed at higher levels or exclusively in U87MG cells. Three proteins, ubiquitin, cystatin B, and tissue transglutaminase (TTG), were upregulated in U87MGdeltaEGFR relative to U87MG. Four proteins highly expressed by U87MG cells, Hsp27, major vault protein, TTG, and cystatin B, were analyzed by Western blot, ELISA, or RT-PCR in cell extracts and in tissue samples of glioblastoma multiforme (GBM; grade IV), low-grade astrocytomas (grades I and II), and nonmalignant brain lesions. All four proteins were highly expressed in GBM tissues compared to nonmalignant brain. These proteins may be used as diagnostic or functional (e.g., multiple drug resistance, invasiveness) markers for glioblastoma tumors. PMID- 12655604 TI - Effects of silkworm paralytic peptide on in vitro hematopoiesis and plasmatocyte spreading. AB - Bombyx mori paralytic peptide (BmPP), a multifunctional cytokine-like molecule, is expressed in the hematopoietic organ-wing imaginal disc complex, suggesting that BmPP is involved in both immune response and the hematopoietic process. We studied the effects of BmPP on plasmatocytes and hematopoietic organs of the silkworm. BmPP (1 microM) stimulated spreading of circulating plasmatocytes, but the percentage of spread plasmatocytes was only 20%. Over 10 nM of BmPP, however, elicited prominent spreading in 70% of young plasmatocytes discharged from cultured hematopoietic organs. Cells in hematopoietic organs that were enzymatically dispersed did not spread even after adding 100 nM of BmPP, indicating that plasmatocytes acquired BmPP-sensitivity immediately after discharge. When cultured in a medium containing larval plasma, hematopoietic organs grew markedly and discharged a large number of hemocytes, over 95% of which were morphologically plasmatocytes. The hemocyte discharge was blocked in the medium containing BmPP dose-dependently, although hematopoietic organ growth was not suppressed. These results suggest that BmPP plays important roles both in hematopoietic regulation and in the hemocyte immune reaction of the silkworm. PMID- 12655605 TI - Yeast-like symbiotes as a sterol source in anobiid beetles (Coleoptera, Anobiidae): possible metabolic pathways from fungal sterols to 7 dehydrocholesterol. AB - Insects are unable to synthesize sterols and require exogenous sterol sources for their normal development and reproduction. A few exceptions are insects associated with symbiotic yeasts or fungi. We analyzed sterols by GC-MS in two anobiid beetles (Lasioderma serricorne and Stegobium paniceum), their intracellular yeast-like symbiotes (YLS), and their diets in order to clarify the sterols synthesized by YLS and the metabolic pathways of the sterols in the beetles. Several C(27), C2(8), and C(29) saturated and unsaturated sterols were identified; the predominant sterols were cholesterol and 7-dehydrocholesterol in the anobiid beetles and ergosterol in the YLS. Most sterols detected in YLS were those known in the late pathway of the ergosterol biosynthesis in yeasts and most of the sterols in the beetles appear to be intermediate metabolites from YLS sterols to 7-dehydrocholesterol. The anobiid beetles appear to use ergosterol and 5-dihydroergosterol as sources for 7-dehydrocholesterol. PMID- 12655606 TI - Eicosanoid involvement in the regulation of behavioral fever in the desert locust, Schistocerca gregaria. AB - The desert locust Schistocerca gregaria behaviorally thermoregulates in order to try and maintain a favoured "set point" body temperature. Locusts infected with the deuteromycete fungal pathogen Metarhizium anisopliae var acridumchoose a significantly elevated temperature. This "behavioral fever" greatly delays the progress of mycosis. We have confirmed this phenomenon and shown that desert locusts also fever when infected with the bacterial pathogen Serratia marcescens. Elevation in the prefered environmental temperature occurs also upon injection with laminarin and lipopolysaccharide (microbial cell wall components). Since such treatments also stimulate the immune system it would appear that "behavioral fever" is probably a feature of the immune response. The eicosanoid biosynthesis inhibitor dexamethasone prevented laminarin invoked fever. This effect was reversable by arachidonic acid. Therefore in common with the febrile response in mammals behavioral fever in insects may be mediated locally by circulating eicosanoids. PMID- 12655607 TI - Development of SCAR markers for the DNA-based detection of the Asian long-horned beetle, Anoplophora glabripennis (Motschulsky). AB - DNA markers were identified for the molecular detection of the Asian long-horned beetle (ALB), Anoplophora glabripennis (Mot.), based on sequence characterized amplified regions (SCARs) derived from random amplified polymorphic DNA (RAPD) fragments. A 2,740-bp DNA fragment that was present only in ALB and not in other Cerambycids was identified after screening 230 random primers in a PCR-based assay system. Three pairs of nested 22-mer oligonucleotide primers were designed on the basis of the sequence of this fragment and were used to perform diagnostic PCR. The first pair of primers (SCAR1) amplified a single 745-bp fragment of ALB DNA, but this did not differentiate ALB from other species. The other two pairs of SCAR primers (SCAR2 and SCAR3) amplified bands of 1,237- and 2,720-bp, respectively, that were capable of differentiating ALB from other closely related non-native and native Cerambycids, such as A. chinensis (Forster), A. malasiaca (Thomson), A. nobilis (Ganglbauer), Monochamus scutellatus (Say), Plectrodera scalator (Fab), Saperda tridentata (Olivier), and Graphisurus fasciatus (Degeer). The latter two SCAR markers could be amplified using DNA extracted from body parts of ALB such as the wing, the leg, and the antennae as well as tissues from all the developmental stages including the egg, larva, pupa, and adult. These markers were also capable of identifying ALB using the DNA extracted from frass. Our results demonstrate that the SCAR markers we have identified can be used for unambiguously identifying ALB from other closely related Cerambycids using a simple PCR procedure. PMID- 12655608 TI - On the ontogeny of Leptodora kindtii (Crustacea, Branchiopoda, Cladocera), with notes on the phylogeny of the Cladocera. AB - Leptodora kindtii, a large predaceous cladoceran, is among the most deviant species of the Cladocera. Therefore, its phylogenetic position has traditionally proven difficult to determine. Its many peculiar features include, among others, long, stenopodous, forwardly directed trunk limbs, a posteriorly placed dorsal brood pouch, a tri-lobed lower lip, and a long, segmented abdomen. This study describes the ontogeny of L. kindtii (Haplopoda), including general body proportions, appendages, the carapace, and other external structures in an attempt to facilitate the comparison of its aberrant morphology to that of other branchiopods. In general, the early embryos are similar to the early embryos of other cladoceran taxa with respect to body shape and size and position and orientation of the early limb buds. Many of the unusual features of L. kindtii appear late in ontogeny. The carapace appears at an early stage as a pair of dorsolateral swellings in a position corresponding to the gap between the mandibles and the first pair of trunk limbs; it later becomes posteriorly transposed by a gradual fusion of its more anterior parts to the dorsal side of the thorax. The tri-lobed "lower lip," under the labrum of the late embryo and the adult, develops as a fusion of the first maxillae (lateral lobes) to an elevated sternal region behind the mouth (median lobe). The stenopodous, segmented trunk limbs in the adult develop from embryonic, elongate, subdivided limb buds, similar to those seen in early stages of other branchiopods. Two conflicting possibilities for the phylogeny of the Cladocera, involving two different positions of L. kindtii (Haplopoda), are discussed. Several characters support a sister-group relationship between the Haplopoda and Onychopoda. However, some characters support the Anomopoda and Onychopoda as sister groups, leaving the Haplopoda outside this clade. In contrast to recent suggestions, we prefer to retain the term "Cladocera" in its original sense as comprising the Haplopoda, Ctenopoda, Anomopoda, and Onychopoda. PMID- 12655609 TI - Formation of the arthrodial membrane in the blue crab, Callinectes sapidus. AB - In this study the pattern of arthrodial membrane deposition in Callinectes sapidus was determined by histological and ultrastructural examination of tissues from the carpus joint of the cheliped collected during premolt, ecdysis, postmolt, and intermolt. Apolysis in the arthrodial membrane occurs at stage D(0) and is synchronous with apolysis of the calcified cuticle. Epicuticle formation begins at early stage D(1) and is completed in late stage D(1). Procuticle deposition starts at D(2) and continues until ecdysis. Numerous cytoplasmic extensions occur throughout the lamellae. Component fibers of the arthrodial membrane are intimately associated with dense plaques on the apical membrane of the underlying hypodermal cells, suggesting a site for fiber polymerization. Deposition of the arthrodial membrane continues after ecdysis, with most of the cuticle thickening occurring during stage C. When stained with PAS and counterstained with hematoxylin, a difference can be discerned between preecdysial and postecdysial procuticle of the arthrodial membrane, a distinction not made in previous studies. The boundary between the arthrodial membrane and calcified cuticle is thicker than either of the two layers and the layers overlap rather than butting up against one another. This pattern suggests that underlying hypodermal cells have to produce multiple types of cuticle over the molt cycle. A summary of the various molting patterns in C. sapidus suggests that the control of these diverse events may prove to be complex. PMID- 12655610 TI - Functional morphology of prey capture in the sturgeon, Scaphirhynchus albus. AB - Acipenseriformes (sturgeon and paddlefish) are basal actinopterygians with a highly derived cranial morphology that is characterized by an anatomical independence of the jaws from the neurocranium. We examined the morphological and kinematic basis of prey capture in the Acipenseriform fish Scaphirhynchus albus, the pallid sturgeon. Feeding pallid sturgeon were filmed in lateral and ventral views and movement of cranial elements was measured from video sequences. Sturgeon feed by creating an anterior to posterior wave of cranial expansion resulting in prey movement through the mouth. The kinematics of S. albus resemble those of other aquatic vertebrates: maximum hyoid depression follows maximum gape by an average of 15 ms and maximum opercular abduction follows maximum hyoid depression by an average of 57 ms. Neurocranial rotation was not a part of prey capture kinematics in S. albus, but was observed in another sturgeon species, Acipenser medirostris. Acipenseriformes have a novel jaw protrusion mechanism, which converts rostral rotation of the hyomandibula into ventral protrusion of the jaw joint. The relationship between jaw protrusion and jaw opening in sturgeon typically resembles that of elasmobranchs, with peak upper jaw protrusion occurring after peak gape. PMID- 12655611 TI - Comparison of testes structure, spermatogenesis, and spermatocytogenesis in young, aging, and hybrid cichlid fish (Cichlidae, Teleostei). AB - Testis structure, spermatogenesis, and spermatocytogenesis were compared in 13 species of cichlid fishes, belonging to the subfamilies Haplochrominae and Tilapinae. The species studied were either mouth brooders, in which fertilization occurs mostly inside the mouth of the brooding fish, or substrate brooders, whose eggs adhere to a substrate over which the sperm is ejaculated. In this study, the embryogenesis of testes anlagen and sperm production was followed in embryos and in fish up to 15 years old, as well as in hybrids of the two subfamilies. In cichlids, the testes are of the unrestricted type and primary spermatogonia develop along the entire length of the developing sperm tubule. The first primary spermatogonia are observed in the testes anlagen 2-5 days after fertilization and they continue to develop in cysts formed by the enveloping Sertoli cells and the intertubular elements. The dimensions of such primary and secondary spermatocysts are correlated with the number of spermatogonia they contain and the corresponding number of mitotic multiplications. The largest mature cysts attained 300 microm, and contained 2,200-2,400 spermatids in the mouth-brooding species and 2,600-3,200 in the substrate-brooding species. Despite the fact that in such cysts cytoplasmic bridges connect only the isogamete spermatids, the maturation of all cells and consequent spermiation is synchronized. Meristic characters distinguish the sperm of mouth brooders from those of substrate brooders, especially in the number of mitochondria and length of the flagellum. In older fish and hybrids, various changes can be seen in the gametogenic epithelium and intertubular cells. These include thickening of the connective tissue, formation of "yellow" groups of Leydig cells, cell apoptosis and degeneration, and, especially, formation of large spermatogonia, with large, electron-dense nucleoli, that have the cytological characteristics of oocytes. The intra- and interspecific variability of sperm dimensions in the studied cichlids poses an interesting question in the context of sperm competition. PMID- 12655612 TI - Surface glycoconjugates in the olfactory system of Ambystoma mexicanum. AB - Lectin binding histochemistry was performed on the olfactory system of neotenic and metamorphosed Ambystoma mexicanum to investigate the distribution and density of defined carbohydrate residues on the cell surface glycoproteins of the olfactory and vomeronasal receptor cells and their terminals in the olfactory bulbs. The lectin binding patterns indicate that the main olfactory system possesses a high density of N-acetyl-galactosamine and N-acetyl-glucosamine residues, while the vomeronasal system contains a high density of N-acetyl galactosamine and galactose moieties and a moderate density of N-acetyl glucosamine. The presence of specific glycoproteins, whose terminal sugars are detected by lectin binding, might be related to the chemoreception and transduction of the odorous message into a nervous signal or to the histogenesis and development of the olfactory system. In fact, the olfactory and vomeronasal receptor cells are neurons that undergo a continuous cycle of proliferation not only during development but also in mature animals. PMID- 12655613 TI - Compartmental fasciotomy and isolating a muscle from neighboring muscles interfere with myofascial force transmission within the rat anterior crural compartment. AB - Muscles within the anterior crural compartment (extensor digitorum longus, EDL; tibialis anterior, TA; and extensor hallucis longus, EHL) and within the peroneal compartment were excited simultaneously and maximally. All muscles were kept at constant length with the exception of EDL, for which muscle length was changed by moving its proximal tendon. Active and passive force was measured at proximal as well as distal EDL tendons and at the combined distal tendons of TA and EHL (TA+EHL). In the initial experimental condition, a difference (F(proximal) > F(distal)) in EDL force, amounting to 0-14% of proximal force, was confirmed for most EDL lengths. This is interpreted as a clear proof of extramuscular myofascial force transmission, as no significant EDL length effects could be shown on TA+EHL force. Repeated measurements were confirmed to cause marked changes of both proximal and distal length-force characteristics, such as a shift of the whole ascending limb of the active curve, including optimum length, to higher lengths without decreasing optimum force, and decreasing active force at low lengths (by approximately 57%). Repeated measurements also lowered proximal and distal EDL passive force (by up to 35%). The proximo-distal difference in passive as well as active EDL force was decreased, but persisted. At most lengths, this difference for active force amounted to a constant fraction (14%) of proximal force. TA+EHL force was not affected significantly. Subsequently, acute effects of experimental surgical alterations were studied: The first manipulation was full lateral fasciotomy of the anterior crural compartment that caused a further decrease in active force at the proximal EDL but not at the distal EDL tendon. Passive forces showed no further significant changes. The proximo-distal EDL active force difference decreased to 0-5% of proximal force. After fasciotomy, TA+EHL force increased by 30%. This was interpreted as evidence of increased intramuscular and decreased extramuscular myofascial force transmission. The second manipulation was full isolation of EDL from TA+EHL, but not from extramuscular connective tissues, which caused a further decrease of the EDL proximo-distal force differences, indicating a stiffening effect of the presence of TA+EHL on the extramuscular matrix. For EDL active force the difference was no longer significantly different from zero. In contrast, for EDL passive force the proximo-distal force difference persisted. It is concluded that extramuscular myofascial force transmission is an important feature of the anterior crural compartment. The magnitude of this force transmission requires that it be considered in analysis of muscular function. PMID- 12655614 TI - Postcanine dental form in the mustelidae and viverridae (Carnivora: Mammalia). AB - This study investigates whether the gross morphology of mustelid and viverrid postcanine dentitions corresponds with differences in diet. For each species, the predominant foods ingested are used to form predictions of dental form and measurements of the carnassial and molar teeth determine the extent of shearing and crushing surfaces on the postcanine teeth. Principal components analysis distinguishes species according to morphological differences in the dentition and these differences are compared with predictions of dental form based on diet. Dietarily specialized species are more likely to be correspondingly specialized in the dentition and species with varied food sources are more likely to possess dental characteristics that are generalized in function. Consumers of foods with high fracture resistance, such as vertebrate tissue and hard-surfaced invertebrates, possess specialized shearing or crushing postcanine teeth. On the other hand, species that consume foods of lesser fracture resistance, such as fruit and soft invertebrates, differ greatly in dental form and are more generalized in dental function. A few species possess postcanine dentitions that do not correspond with diet; the absence of dental-dietary correlation in these species suggests that other factors, such as phylogeny, are important in determining dental form. PMID- 12655615 TI - Craniofacial variation of the Chinese macaques explored with Morphologika. AB - In order to analyze, separately and dynamically, the variation in cranial size and shape of Chinese macaques, a new method, Morphologika, was used to illustrate 3D profiles based on the coordinates of 26 landmarks on the skull. Striking image variation between the two sexes was detected on the facial region: males exhibited a larger and more protrusive facial structure. Males also displayed a bigger cranium than females. The two sexes also showed quite different images in skull shape. However, they expressed the same allometric pattern with regard to the relationship between size and shape, which was significantly positively associated with each other along the first axis. The same relationship was negatively displayed along the second axis when the two sexes were analyzed together. However, only the relationship for females reached a significant level when the two sexes were studied separately. This was considered to be related to their differentiation in growth trajectory. This study also tested the concept of size revealed by the second axis of principal components analysis based on traditional morphometric methods on the same taxa. PMID- 12655616 TI - Altered histology of the thymus and spleen in contaminant-exposed juvenile American alligators. AB - Morphological differences in spleen and thymus are closely related to functional immune differences. Hormonal regulation of the immune system has been demonstrated in reptilian splenic and thymic tissue. Spleens and thymus were obtained from juvenile alligators at two reference sites in Florida, USA: Orange Lake and a National Wildlife Refuge, Lake Woodruff, as well as from a contaminated lake, Lake Apopka. Lake Apopka has been extensively polluted with agricultural pesticides. Tissues were prepared for histological analysis to determine if previously detected endocrine abnormalities associated with contaminant exposure might also be reflected in morphological differences in splenic and thymic structures important for immunological response. Similar tissues were taken from captive-raised juvenile female alligators (3 years old) that were hatched from eggs collected on Lake Woodruff and Lake Apopka. Differences in thymic ratios (medulla/cortex) were found among alligators collected from the two lakes (P = 0.0051). Alligators from Lake Apopka had smaller thymic ratios than animals from either reference lake. Males from Lake Woodruff had significantly smaller lymphocyte sheaths in the spleen than females (P = 0.0009), indicative of a normal sexual dimorphism. Lymphocyte sheath width differed among females obtained from the three lakes, with females from Lake Apopka having the smallest sheath width and those from Orange Lake having the largest. Malpighian body area was largest in alligators from Orange Lake, intermediate in Lake Woodruff, and smallest in Lake Apopka. In contrast to that observed for wild-caught animals, no difference was found in the thymic medulla/cortex ratio of captive-raised female alligators (P = 0.378). Captive raised female alligators from Lake Apopka and Lake Woodruff displayed lake associated differences in lymphocyte sheath width as observed in wild animals; Lake Apopka alligators had smaller lymphocyte sheath width compared to Woodruff alligators (P = 0.0396). In contrast to wild-caught animals, area of the Malpighian bodies did not differ by lake in the captive-raised female alligators (P = 0.066). The enlarged thymic cortex suggests a change in T-lymphocyte maturation within the thymus of alligators from a contaminated environment, Lake Apopka. The results point to alterations in the histology of the thymus and spleen. Further studies are required to examine the functional significance of these observations. PMID- 12655617 TI - Stomach of Aplysia depilans (Mollusca, Opisthobranchia): a histochemical, ultrastructural, and cytochemical study. AB - We report the results of a morphological, histochemical, and cytochemical characterization of the Aplysia depilans stomach, an organ little studied in opisthobranchs. Very thin ciliated cells with microvilli on their apical surfaces are predominant in the epithelium lining the lumen of the stomach. Many lysosomes with a strong arylsulphatase activity were present in the apical regions of these cells that could also contain some lipid droplets and glycogen. Small peroxisomes were observed, usually around lipid droplets or mitochondria. Bottle-shaped secretory cells are very common in this epithelium and produce a secretion rich in proteins and acidic mucopolysaccharides. Most of the cytoplasm of these mucus producing cells was filled with a very high number of granules and the nucleus is dislocated to the basal region. Cisternae of rough endoplasmic reticulum were abundant around the nucleus and several Golgi stacks were also present in this area. In spite of the variation in the electron density of the granules, only one type of secretory cell seems to be present in the stomach epithelium, since granules with very different electron densities were frequently found in the same cell. A few neurons were also found in the stomach epithelium of this species. Fibrocytes, muscle cells, nerves, and amebocytes were observed in the connective tissue of the stomach wall. PMID- 12655618 TI - Ultrastructural and ultracytochemical study of the middle ear epithelium in the chicken, Gallus gallus domesticus. AB - The fine structure of the epithelium lining the tympanic cavity of the chicken was studied by TEM and SEM. In addition, the distribution of nonspecific esterase activity in the epithelium was investigated by TEM. Ultrastructural study revealed the presence of disk-like apical protrusions of the epithelial cells, previously not observed in other cell types. The protrusions contained some cytoplasmic organelles and were characterized by a ring-shaped thickening around their periphery. The ring was made up of a granulo-filamentous material. Our observations clearly indicate the existence of an apocrine secretory mechanism, consisting of a progressive detachment of disk-like protrusions from the apex of the epithelial cells. The ultracytochemical study demonstrated nonspecific esterase activity on the epithelial surface and in the secretory vesicles. We propose that nonspecific esterase is a marker for middle ear surfactant in birds. PMID- 12655619 TI - Comparison of group and individual cognitive-behavioral therapy for patients with bulimia nervosa. AB - OBJECTIVE: The clinical effectiveness of group and individual cognitive behavioral therapy (CBT) for bulimia nervosa (BN) was compared. METHOD: Sixty BN patients from hospitals and general practitioners in Sydney, Australia, were allocated randomly to group or individual CBT. Forty-four completed treatment (n = 22 in group CBT and n = 22 in individual CBT). Patients were assessed at pretreatment, posttreatment, and at 3 and 6 months follow-up with the Eating Disorder Examination-12 and self-report questionnaires examining weight and shape attitudes (Eating Disorder Inventory-2), social adjustment (Social Adjustment Scale-Modified), self-esteem (Rosenberg Self-Esteem Scale), and general psychopathology (Symptom Checklist 90R). RESULTS: The effects of group and individual CBT were equivalent on most measures. However, a significantly greater proportion of individual CBT patients than group CBT patients were abstinent from bulimic behaviors at posttreatment, but not at follow-up. DISCUSSION: This has implications for the delivery of cost-effective and clinically effective treatment for BN. PMID- 12655621 TI - Anxiolytic effects of acute tryptophan depletion in anorexia nervosa. AB - OBJECTIVE: Recent studies have raised the question as to whether a dysregulation of the neurotransmitter serotonin may contribute to the alterations in mood seen in anorexia nervosa (AN). People with AN tend to be anxious, obsessional, perfectionistic, and harm avoidant. These traits are premorbid and persist after recovery. It has been suggested that increased activity of brain serotonin systems could contribute to this pathologic condition. Dieting in AN, which serves to reduce plasma levels of tryptophan (TRP), may serve to reduce symptoms of dysphoric mood. METHOD: Fourteen women currently symptomatic with AN (ILL AN), 14 women recovered from AN (REC AN), and 15 healthy control women (CW) underwent acute tryptophan depletion (ATD). Measures of psychological state were self assessed at baseline and hourly after ATD to determine whether ATD would reduce negative mood. RESULTS: ILL AN and REC AN had significantly higher mean baseline TRP/LNAA (tryptophan/large neutral amino acids) ratios compared with CW. In contrast to placebo, the ATD challenge demonstrated a significantly greater reduction in the TRP/LNAA ratio for ILL AN (-95%) and REC AN (-84%) compared with CW (-70 %). Both the ILL AN and REC AN had a significant reduction in anxiety on the ATD day compared with the placebo day. DISCUSSION: These data demonstrate that a dietary-induced reduction of TRP, the precursor of serotonin, is associated with decreased anxiety in people with AN. Restricting dietary intake may represent a mechanism through which individuals with AN modulate a dysphoric mood. PMID- 12655623 TI - Neuroticism and low self-esteem as risk factors for incident eating disorders in a prospective cohort study. AB - OBJECTIVE: To assess the role of neuroticism and low self-esteem as risk factors for eating disorders (ED). METHOD: A representative sample of girls 12-21 years old from Navarre, Spain. Girls free from any ED in 1997 were followed up for 18 months and reevaluated using DSM-IV criteria. Multivariable logistic models were used to examine associations between neuroticism (Eysenck Inventory) or low self esteem (36-item scale) and incident ED. RESULTS: Higher levels of neurotic personality increased the risk of ED (adjusted odds ratio [OR] for the highest quartile, 3.3; 95% Cl; 1.6-4.8). High levels of self-esteem were protective (OR, 0.32; 95% CI, 0.16-0.66). Neuroticism was a more powerful predictor than low self esteem. DISCUSSION: Our results provide prospective evidence supporting the role of neuroticism and low self-esteem as major determinants of ED. PMID- 12655624 TI - A dialectical behavior therapy program for people with an eating disorder and borderline personality disorder--description and outcome. AB - OBJECTIVE: To describe and evaluate a full dialectical behavior therapy (DBT) program for people with comorbid eating disorder and borderline personality disorder. The program included a novel skills training module written especially for eating-disordered patients. METHOD: The program was run for 18 months. Days in hospital and major acts of self-harm were counted for the 18 months before and after DBT. RESULTS: There were no dropouts from the program. The patients seemed to benefit. Most patients were neither eating disordered nor self-harming at follow-up. DISCUSSION: Full DBT is an expensive and demanding treatment but deserves consideration for patients with an eating disorder and co-morbid borderline personality disorder and self-harm. There is a need for a more systematic and thorough evaluation. PMID- 12655625 TI - Differential heritability of eating attitudes and behaviors in prepubertal versus pubertal twins. AB - OBJECTIVE: Differential heritability of eating pathology in prepubertal, pubertal, and 17-year-old twins was examined using a population-based twin sample. METHOD: Participants included 530 11-year-old (452 prepubertal, 78 pubertal twins) and 602 17-year-old female twins from the Minnesota Twin Family Study. Puberty was assessed with the Pubertal Development Scale, and general eating pathology was assessed with the total score from the Minnesota Eating Disorders Inventory (M-EDI). RESULTS: M-EDI total score intraclass correlations for the pubertal 11-year-old group were more similar to the 17-year-old twins' than the prepubertal 11-year-old twins, suggesting similar levels of genetic influence in the pubertal and older adolescent cohort. Model-fitting corroborated these impressions, indicating that genetic factors accounted for 0% of the variance in 11-year-old prepubertal twins but 54% of the variance in 11-year-old pubertal and 17-year-old twins. DISCUSSION: Findings suggest that a portion of the genetic influence on eating pathology may become activated during puberty. PMID- 12655626 TI - Genetic and environmental contributions to obesity and binge eating. AB - OBJECTIVE: Binge eating is present in a substantial proportion of obese individuals. A tendency toward obesity has been identified as a risk factor for eating disorders such as bulimia nervosa and binge eating disorder. The purpose of this article was to determine the extent of overlap between genetic and environmental factors that contribute to the liability to obesity and binge eating. RESEARCH METHOD: In a population-based sample of 2163 female twins, we conducted bivariate twin modeling to explore the relation between the genetic and environmental risk factors for obesity and binge eating. RESULTS: Bivariate twin modeling revealed substantial heritability for obesity (0.86: 95% CI, 0.77-0.94), moderate heritability for binge eating (0.49: 95% CI, 0.38-0.61), and a modest genetic correlation of +.34 (95% CI, 0.19-0.50) between the two traits. CONCLUSIONS: Both binge eating and obesity are heritable conditions, and there seems to be only modest overlap in the genetic risk factors that increase liability to each condition. PMID- 12655627 TI - Cognitive processing of anorexic patients in recognition tasks: an event-related potentials study. AB - OBJECTIVE: We used event-related potentials (ERPs) to test whether anorexic subjects have difficulties in filtering out irrelevant stimuli in controlled information processing tasks. METHODS: ERPs from 12 anorexic patients were recorded during recognition of simple and complex body images and simple and complex geometrical shapes. RESULTS: Anorexic subjects had larger P300 amplitudes for frequent stimuli during body images and simple geometrical shape recognition tasks. Longer P300 latencies were also found in simple geometrical shape recognition tasks, although task complexity had no effect on the P300 latency and amplitude. DISCUSSION: These results are explained in terms of nonspecific hyperarousal in mental anorexia and relative inability to filter out irrelevant stimuli leading to working memory saturation. PMID- 12655628 TI - Obsessions and compulsions in anorexia nervosa subtypes. AB - OBJECTIVE: Obsession and compulsions in anorexia nervosa (AN) patients are often confused with the preoccupations and rituals that are characteristic of obsessive compulsive disorder (OCD). We examined the type and frequency of characteristic OCD obsessions and compulsions in a large sample of AN patients. METHOD: In personal interviews with 324 AN patients, we assessed lifetime histories of eating disorder symptomatology and obsessive-compulsive behaviors with valid semistructured interviews. Checklist category sums on the Yale-Brown Obsessive Compulsive Scale were compared between AN and OCD subjects using generalized estimating equations. RESULTS: Lifetime obsessions and compulsions occurred in 68% of the AN restricting type and in 79.1% of the AN binge/purge type. The AN subgroups did not differ from OCD controls in frequency of obsessions in the symmetry and somatic categories or in the compulsion categories of ordering and hoarding. In all other categories, the AN subgroups had a significantly lower frequency compared with the OCD controls. DISCUSSION: Some common phenotype characteristics shared by most AN and OCD patients suggest these disorders may share common brain behavioral pathways. However, the lack of complete overlap indicates they most likely have different loci of pathology within those pathways. PMID- 12655629 TI - A longitudinal examination of patterns in girls' weight concerns and body dissatisfaction from ages 5 to 9 years. AB - OBJECTIVE: To examine the development of girls' weight concerns and body dissatisfaction from ages 5 to 9 years and how change and continuity in these constructs from age 5 to 7 years is associated with girls' eating attitudes, dietary restraint, and dieting status at age 9. METHODS: Weight concerns, body dissatisfaction, and weight status were assessed for 182 girls when they were 5, 7, and 9 years old, and their eating attitudes, dietary restraint, and dieting status were assessed when they were 9. RESULTS: Girls tended to maintain their rank in weight concerns and body dissatisfaction across ages 5 to 9 years, and associations among girls' weight concerns, body dissatisfaction, and weight status increased with age. In addition, positive associations were found between changes in girls' weight concerns, body dissatisfaction, and weight status across ages 7 to 9. Finally, girls' who reported high weight concerns or high body dissatisfaction across ages 5 to 7 reported higher dietary restraint, more maladaptive eating attitudes, and a greater likelihood of dieting at age 9, independent of their weight status. CONCLUSION: Girls' reported weight concerns and body dissatisfaction across middle childhood showed consistency over time, were systematically related to their weight status, and predicted their dietary restraint, eating attitudes, and the likelihood of dieting at age 9. These results reflect patterns identified among adolescent girls and women. PMID- 12655630 TI - The nature of body image disturbance in patients with binge eating disorder. AB - OBJECTIVE: This study examined the distinction between body dissatisfaction and self-evaluation unduly influenced by body shape and weight, and their longitudinal relationships to depressive symptomatology and self-esteem in patients with binge eating disorder (BED). METHOD: Ninety-seven patients with BED completed measures tapping these constructs at baseline and again 4 weeks later. RESULTS: Change in body dissatisfaction was significantly correlated with both change in depressive symptomatology and change in self-esteem over time, whereas change in self-evaluation was significantly correlated only with change in self esteem. In addition, change in shape concern, but not change in weight concern, was significantly correlated with change in self-esteem only. DISCUSSION: These findings suggest that self-evaluation unduly influenced by body shape is a more useful indicator for BED than body dissatisfaction or self-evaluation unduly influenced by weight. PMID- 12655631 TI - Body image dissatisfaction and disordered eating in black and white women. AB - OBJECTIVE: This study predicted and found that body image dissatisfaction and bulimic symptoms have a curvilinear relationship among undergraduate women. RESULTS: For the women in this sample, regardless of race, body image dissatisfaction correlated with bulimic symptoms, such that women who perceived themselves as bigger or smaller than the ideal body size for their ethnic group endorsed bulimic symptoms. Black and white women differed regarding their ethnic group's ideal body image and their self-perceptions of how they compared with the ideal image. Black women tended to report being underweight, whereas white women tended to report being overweight. DISCUSSION: The findings in this study suggest that some black women are not buffered against eating disorders as suggested in previous research. PMID- 12655632 TI - Cognitive impairment in anorexia nervosa is not due to depressed mood. AB - OBJECTIVE: Previous research has revealed cognitive deficits in patients with anorexia nervosa. It is unclear whether these deficits are linked to co-morbid depression in this population. We examined the relationship between depressive symptoms and cognitive functioning. METHOD: A large sample of subjects (N = 98) was examined using a common measure of cognitive ability and two measures of depression. RESULTS: We confirmed that there is no relationship between depression and cognitive ability in this group. DISCUSSION: This suggests that alternative explanations be explored for cognitive deficits in patients with anorexia nervosa. PMID- 12655633 TI - Achalasia mimicking prepubertal anorexia nervosa. AB - A 9-year-old girl presents for continuing weight loss of 10 kg over the course of 1 year. Medical history showed three episodes of pneumonia requiring hospital admission in the 6 months before presentation and 4 months of weekly psychotherapy for anorexia nervosa. A thorough history of eating behavior and a review of systems revealed not only typical aspects of prepubertal anorexia nervosa but also vomiting at night while asleep, difficulty drinking liquids, epigastric pain, and a frequent experience of "a lump in the throat"; these symptoms were not suggestive of a diagnosis of anorexia nervosa but rather of esophageal achalasia. The patient was transferred to the Department of Pediatrics, and a diagnosis of esophageal achalasia was made by chest x-ray and barium swallow. After dilatation and botulinum toxin application, the patient regained weight easily and was discharged in stable condition. In this case, esophageal achalasia mimicked prepubertal anorexia nervosa. PMID- 12655634 TI - Severe neutropenia with gelatinous bone marrow transformation in anorexia nervosa: a case report. AB - A 28-year-old woman with a history of anorexia nervosa was admitted with excessive weight loss, edema, and amenorrhea. She had lost 34% of her previous body weight within 2 years, and her body mass index was 12.3 kg/m(2). The leukocyte count on admission was 2150/microl and gradually decreased to 980/microl (neutrophils; 276/microl). Bone marrow biopsy disclosed gelatinous transformation with hypocellularity. After the patient was treated with intravenous nutritional support, the severe neutropenia improved to the level on admission. Hematological abnormalities seem to be common in anorexia nervosa, but severe neutropenia with gelatinous bone marrow transformation has rarely been reported. PMID- 12655636 TI - A small issue addressed. AB - Cell size is an important determinant of body size. While the genetic mechanisms of cell size regulation have been well studied in yeast, this process has only recently been addressed in multicellular organisms. One recent report by Wang et al. (2002) shows that in the nematode C. elegans, the TGFbeta-like pathway acts in the hypodermis to regulate cell size and consequently body size.1 This finding is an exciting step in discovering the molecular mechanisms that control cell and body size. PMID- 12655635 TI - Echocardiographic investigation of pericardial effusion in a case of anorexia nervosa. AB - OBJECTIVE: Pericardial effusion has recently been reported as a complication of anorexia nervosa. A distinct pathophysiological cause of it could not be revealed. In some reports, there was a probable correlation between weight gain and reduction of pericardial effusion in anorexia nervosa cases. We encountered a case in which pericardial effusion remitted completely along with body weight increase and normalization of low T3 syndrome. These findings suggest that the reduction of pericardial effusion may correlate with both weight gain and low T3 normalization. Plasma brain natriuretic peptide (BNP) levels were increased in this case despite heart failure, and plasma BNP decreased as pericardial effusion remitted. The measurement of serum BNP level may be a clinical parameter in such a case of pericardial effusion. PMID- 12655637 TI - Topoisomerase II may be linked to the reduction of chromosome number in meiosis. AB - A reduction of chromosome number in meiosis is essential for genome transmission in diploid organisms. Reduction depends on a change in kinetochore configuration.1 A recent study2 connects changes in kinetochores with other changes in chromosome structure and raises the intriguing possibility that topoisomerase II, the DNA untangling enzyme, is involved. PMID- 12655638 TI - Block to DNA replication in meiotic maturation: a unified view for a robust arrest of cell cycle in oocytes and somatic cells. AB - Under certain conditions, the cell cycle can be arrested for a long period of time. Vertebrate oocytes are arrested at G(2) phase, while somatic cells arrest at G(0) phase. In both cells, nuclei have lost the ability to initiate DNA synthesis. In a pair of recently published papers,[1,2] Mechali and colleagues and Coue and colleagues have clarified how frog oocytes prevent untimely DNA synthesis during the long G(2) arrest. Intriguingly, they found only Cdc6 is responsible for the inability of immature oocytes to replicate DNA. Cdc6 is a key component for replication licensing, and for G(0) cells to re-enter the proliferative stage. Strikingly similar strategies for preventing the untimely replication in both cells suggest that the suppression of replication licensing is a universal mechanism for securing the prolonged arrest of the cell cycle. PMID- 12655639 TI - Wnt-frizzled signaling in the induction and differentiation of the neural crest. AB - The neural crest is a transient population of multipotent progenitors arising at the lateral edge of the neural plate in vertebrate embryos. After delamination and migration from the neuroepithelium, these cells contribute to a diverse array of tissues including neurons, smooth muscle, craniofacial cartilage, bone cells, endocrine cells and pigment cells. Considerable progress in recent years has furthered our understanding at a molecular level of how this important group of cells is generated and how they are assigned to specific lineages. Here we review a number of recent studies supporting a role for Wnt signaling in neural crest induction, differentiation, and apoptosis. We also summarize the timing of expression of a number of Wnt ligands and receptors with respect to neural crest induction. PMID- 12655640 TI - Transcriptional silencing and translational control: key features of early germline development. AB - The germ lineage has been studied for a long time because of its crucial role in the propagation and survival of a species. While this lineage, in contrast to the soma, is clearly unique in its totipotent ability to produce a new organism, it has now been found also to have specific features at the cellular level. One feature, a period of transcriptional quiescence in the early germ cell precursors, has been observed in both Drosophila and C. elegans, where it is essential for the formation and the survival of the germline. In addition, there are numerous instances where these early germ cells are reliant on translational regulation, especially in Drosophila. The genes that are important for these two functions, the mechanisms of their action, and studies in vertebrate organisms that reveal similarities as well as some potential differences in early germ cell development are discussed. PMID- 12655641 TI - Actin-based motility: from molecules to movement. AB - Extensive progress has been made recently in understanding the mechanism by which cells move and extend protrusions using site-directed polymerization of actin in response to signalling. Insights into the molecular mechanism of production of force and movement by actin polymerization have been provided by a crosstalk between several disciplines, including biochemistry, biomimetic approaches and computational studies. This review focuses on the biochemical properties of the proteins involved in actin-based motility and shows how these properties are used to generate models of force production, how the predictions of different theoretical models are tested using a biochemically controlled reconstituted motility assay and how the changes in motility resulting from changes to the concentrations of components of the assay can help understand diverse aspects of the motile behavior of living cells. PMID- 12655642 TI - Neurofilaments and neurological disease. AB - Neurofilaments are one of the major components of the neuronal cytoskeleton and are responsible for maintaining the calibre of axons. They are modified by post translational changes that are regulated in complex fashions including by the interaction with neighbouring glial cells. Neurofilament accumulations are seen in several neurological diseases and neurofilament mutations have now been associated with Charcot-Marie-Tooth disease, Parkinson's disease and amyotrophic lateral sclerosis. In this review, we discuss the structure, normal function and molecular pathology of neurofilaments. PMID- 12655643 TI - The MID1/PP2A complex: a key to the pathogenesis of Opitz BBB/G syndrome. AB - Opitz BBB/G syndrome is a monogenic disorder that is characterized by malformations of the ventral midline. Investigations into the underlying genetic defects and the pathobiochemistry of this syndrome have already shed light on the mechanisms of both the physiological and the pathological development of the ventral midline, a complicated multistep process. Moreover, these studies have revealed the ubiquitin-dependent regulation of microtubule-associated phosphatase 2A, a central mechanism in many cellular processes. In this review, we summarize recent findings and speculate upon their implications for both medical and general research. PMID- 12655644 TI - Enzymes and auxiliary factors for GPI lipid anchor biosynthesis and post translational transfer to proteins. AB - GPI lipid anchoring is an important post-translational modification of eukaryote proteins in the endoplasmic reticulum. In total, 19 genes have been directly implicated in the anchor synthesis and the substrate protein modification pathway. Here, the molecular functions of the respective proteins and their evolution are analyzed in the context of reported literature data and sequence analysis studies for the complete pathway (http://mendel.imp.univie.ac.at/SEQUENCES/gpi-biosynthesis/) and questions for future experimental investigation are discussed. Studies of two of these proteins have provided new mechanistic insights. The cytosolic part of PIG-A/GPI3 has a two-domain alpha/beta/alpha-layered structure; it is suggested that its C terminal subsegment binds UDP-GlcNAc whereas the N-terminal domain interacts with the phosphatidylinositol moiety. The lumenal part of PIG-T/GPI16 apparently consists of a beta-propeller with a central hole that regulates the access of substrate protein C termini to the active site of the cysteine protease PIG K/GPI8 (gating mechanism) as well as of a polypeptide hook that embraces PIG K/GPI8. This structural proposal would explain the paradoxical properties of the GPI lipid anchor signal motif and of PIG-K/GPI8 orthologs without membrane insertion regions in some species. PMID- 12655645 TI - Trilobite body patterning and the evolution of arthropod tagmosis. AB - Preservation permitting patterns of developmental evolution can be reconstructed within long extinct clades, and the rich fossil record of trilobite ontogeny and phylogeny provides an unparalleled opportunity for doing so. Furthermore, knowledge of Hox gene expression patterns among living arthropods permit inferences about possible Hox gene deployment in trilobites. The trilobite anteroposterior body plan is consistent with recent suggestions that basal euarthropods had a relatively low degree of tagmosis among cephalic limbs, possibly related to overlapping expression domains of cephalic Hox genes. Trilobite trunk segments appeared sequentially at a subterminal generative zone, and were exchanged between regions of fused and freely articulating segments during growth. Homonomous trunk segment shape and gradual size transition were apparently phylogenetically basal conditions and suggest a single trunk tagma. Several derived clades independently evolved functionally distinct tagmata within the trunk, apparently exchanging flexible segment numbers for greater regionally autonomy. The trilobite trunk chronicles how different aspects of arthropod segmentation coevolved as the degree of tagmosis increased. PMID- 12655646 TI - Evolution of adrenal and sex steroid action in vertebrates: a ligand-based mechanism for complexity. AB - Various explanations have been proposed to account for complex differentiation and development in humans, despite the human genome containing only two to three times the number of genes in invertebrates. Ignored are the actions of adrenal and sex steroids-androgens, estrogens, glucocorticoids, mineralocorticoids, and progestins-which act through receptors that arose from an ancestral nuclear receptor in a protochordate. This ligand-based mechanism is unique to vertebrates and was integrated into the already robust network of transcription factors in invertebrates. Adrenal and sex steroids influence almost all aspects of vertebrate differentiation and development. I propose that evolution of this ligand-based mechanism in a primitive vertebrate was an important contribution to vertebrate complexity. Sequencing of genomes from a cephalochordate, such as amphioxus, and from hagfish and lamprey will establish early events in the evolution of steroid hormone signaling, and also allow genetic studies to elucidate how vertebrate complexity depends on steroid hormones. PMID- 12655647 TI - The intracrine hypothesis and intracellular peptide hormone action. AB - There is evidence that many peptide growth factors and hormones act in the intracellular space after either internalization or retention in their cells of synthesis. These factors, commonly called intracrines, are structurally diverse while sharing some common functional features. Reports of intracellular peptide hormone binding and action are reviewed here. Also, this laboratory has made proposals regarding the origin and actions of intracrines and these areas are further explored. Intracrine interactions and the relationship of intracrines to transcription factors are discussed. The intracellular/intracrine renin angiotensin system (iRAS) is reviewed to illustrate the intracrine analogue of a well-established physiological system. The role of intracrine action in metazoan development is also considered. PMID- 12655648 TI - Eyespot placement and assembly in the green alga Chlamydomonas. AB - The eyespot organelle of the green alga Chlamydomonas allows the cell to phototax toward (or away) from light to maximize the light intensity for photosynthesis and minimize photo-damage. At cytokinesis, the eyespot is resorbed at the cleavage furrow and two new eyespots form in the daughter cells 180 degrees from each other. The eyespots are positioned asymmetrically with respect to the microtubule cytoskeleton. Eyespots are assembled from all three chloroplast membranes and carotenoid-filled granules, which form a sandwich structure overlaid by the tightly apposed plasma membrane. This review describes (1) my interest in cellular asymmetry and organelle biology, (2) isolation of mutations that describe four genes governing eyespot placement and assembly, (3) the characterization of the EYE2 gene, which encodes a thioredoxin superfamily member, and (4) the characterization of the MIN1 gene, which is required for the layered organization of granules and membranes in the eyespot. BioEssays 25:410 416, 2003. PMID- 12655649 TI - Chronology of cellular alterations during 7-ketocholesterol-induced cell death on A7R5 rat smooth muscle cells: analysis by time lapse-video microscopy and conventional fluorescence microscopy. AB - BACKGROUND: Time-lapse video microscopy was used to determine whether mitochondrial and nuclear changes (decrease in mitochondrial transmembrane potential, condensation, and/or fragmentation of the nuclei, morphologic features typical of apoptosis) occurring during 7-ketocholesterol-induced cell death on A7R5 rat smooth muscle cells took place before or after the loss of cell adhesion. In addition, changes in actin organization were followed by conventional fluorescence microscopy. METHODS: Morphologic, functional, and spatial changes at the mitochondrial level were investigated with 3,3' dihexyloxacarbocyanine iodide and/or MitoTracker Red, and nuclear morphology was characterized by staining with Hoechst 33342. Actin fibers, which are major components of the filament network of the cytoskeleton, were visualized with phalloidin linked to fluorescein. The numbers of adherent and nonadherent cells were determined by cell counting. RESULTS: 7-Ketocholesterol-induced cell death was associated with a rapid alteration of actin fibers, a loss of intercellular junctions, and cell shape modifications. Analysis of mitochondrial transmembrane potential showed successively a hyperpolarization and a more or less pronounced progressive decrease followed by a dramatic drop associated with an increase in Hoechst 33342 staining, reflecting chromatin condensation and morphologic changes in the nuclei. CONCLUSIONS: During cell death induced by 7-ketocholesterol in A7R5 rat smooth muscle cells, the different methods of microscopy allowed us to establish that alterations of actin fibers and mitochondrial dysfunctions occurred before condensation and/or fragmentation of the nuclei, which preceded the loss of cell adhesion. PMID- 12655650 TI - DNA content, chromatin texture and nuclear morphology in benzo[a]pyrene transformed human breast epithelial cells after microcell-mediated transfer of chromosomes 11 and 17. AB - BACKGROUND: A relation between the changes in DNA content and chromatin supra organization and the expression of gradual steps of tumorigenesis has been assessed by image analysis in human breast epithelial cells (MCF-10F) treated with benzo[a]pyrene (BP) (cell lines BP1, BP1-E, BP1-Tras, and others). METHODS: Because abnormal chromosomes 11 and 17 have been associated with neoplastic progression in BP-transformed MCF-10F cells, image analysis of Feulgen-stained tumorigenic BP1-E cells with the microcell-mediated chromosome transfer of normal chromosomes 11 and 17 was carried out. RESULTS: A tendency of DNA amount distribution and nuclear size restoration to values typical of non-transformed MCF-10F cells was demonstrated, especially after the transfer of chromosome 17. No reversion in chromatin texture was found after the transfer of chromosome 11 or 17. CONCLUSIONS: Although the presence of a normal chromosome 17 should be considered among the necessary steps for tumorigenic human breast epithelial cells to recover their normality, a more complex genome balance is required for the entire nuclear chromatin of these cells to recover its totally normal supra organization and expression. PMID- 12655651 TI - Fluorescence lifetime-resolved pH imaging of living cells. AB - BACKGROUND: The regulation and maintenance of intracellular pH are critical to diverse metabolic functions of the living cells. Fluorescence time-resolved techniques and instrumentations have advanced rapidly and enabled the imaging of intracellular pH based on the fluorescence lifetimes. METHODS: The frequency domain fluorescence lifetime imaging microscopy (FLIM) and fluorophores displaying appropriate pH-dependent lifetime sensitivities were used to determine the temporal and spatial pH distributions in the cytosol and vesicular compartment lysosomes. RESULTS: We found that cytosolic pH levels are different in 3T3 fibroblasts, Chinese hamster ovary (CHO) cells, and MCF-7 cells when using the pH probe carboxy-SNAFL2. We also tracked the transient cytosolic pH changes in the living CHO cells after treatments with proton pump inhibitors, ion exchanger inhibitors, and weak base and acid. The intracellular lysosomal pH was determined with the acidic lifetime probes DM-NERF dextrans, OG-514 carboxylic acid dextrans, and LysoSensor DND-160. Our results showed that the resting lysosomal pH value obtained from the 3T3 fibroblasts was between 4.5 and 4.9. The increase of lysosomal pH induced by the treatments with proton pump inhibitor and ionophores also were observed in our FLIM measurements. CONCLUSIONS: Our lifetime based pH imaging data suggested that FLIM can measure the intracellular pH of the resting cells and follow the pH fluctuations inside the cells after environmental perturbations. To improve the z-axis resolution to the intracellular lifetime resolved images, we are investigating the implementation of the pseudo-confocal capability to our current FLIM apparatus. PMID- 12655652 TI - Comparison of reagents for shape analysis of fixed cells by automated fluorescence microscopy. AB - BACKGROUND: Cell size and shape have been implicated as potentiators of intracellular signaling events and as indicators of abnormal cell behavior. Automated microscopy and image analysis can provide quantitative information about the size and shape of cultured cells, but it requires that the edge of a cell be clearly identified. Generating adequate contrast at the edge of thin well spread cells can be challenging. METHODS: We compared six (five chemically reactive and one lipophilic) fluorescent molecules--5-chloromethyl fluorescein diacetate (CMFDA, CellTracker green), fluorescein-5-maleimide, fluorescein-5 isothiocyanate (FITC), 5-iodoacetamidofluorescein, 5(6)-carboxy fluorescein-N hydroxysuccinimidyl ester, and N-fluorescein-1,2-dihexadecanoyl-sn-glycerol-3 phosphoethanolamine--for their effectiveness as stains for automated morphology analysis of fixed cells. RESULTS: Formaldehyde-fixed rat aortic smooth muscle cells stained with fluorescein-5-maleimide or FITC exhibited an average intensity that was at least twofold greater than cells stained with CMFDA even when subjected to a 25-fold shorter exposure time. Cell area determined with the higher intensity stains was less sensitive to threshold settings during automated cell morphology analysis. CONCLUSION: A procedure that includes the use of fluorescein-5-maleimide or FITC for staining fixed cell provides sensitivity sufficient to permit rapid, automated, morphologic analysis of well-spread fixed cells. PMID- 12655653 TI - Effect of anti-CD20 monoclonal antibody, Rituxan, on cynomolgus monkey and human B cells in a whole blood matrix. AB - BACKGROUND: Cynomolgus monkeys are widely used animal models in biomedical research. The differences between cynomolgus monkey and human B cells are not completely understood. However, these differences are of crucial importance for interpretation of data from studies on new therapeutic agents aimed at B-cell depletion, such as anti-CD20 monoclonal antibodies. METHODS: Multicolor fluorescence-activated cell sorting analysis of peripheral blood B cells was performed on samples treated ex vivo with the anti-CD20 therapeutic monoclonal antibody, Rituxan, in a whole blood matrix. RESULTS: In contrast to humans, cynomolgus monkeys had two distinct B-cell subsets, CD20highCD40lowCD21- and CD20lowCD40highCD21+. These B-cell subsets had a 2.5-fold difference in the EC50 for Rituxan binding and differed significantly in their in vitro susceptibility to Rituxan depletion. Human B cells were similar to the CD20lowCD40highCD21+ cynomolgus monkey B cells with regard to their EC50 for Rituxan and response to Rituxan in a whole blood matrix assay. CD21 was upregulated, whereas CD40 was downregulated at incubation with Rituxan in the CD20lowCD40highCD21+ monkey and human B cells in a concentration-dependent manner. CONCLUSIONS: These findings have direct implications for in vivo studies of therapeutic agents that target B cells in cynomolgus monkeys and for extrapolation of the results to humans. In addition, our data are consistent with the model in which CD20, CD21, and CD40 exist in a supramolecular complex that is affected by anti-CD20 monoclonal antibodies. PMID- 12655654 TI - Comparison of the effect of mitochondrial inhibitors on mitochondrial membrane potential in two different cell lines using flow cytometry and spectrofluorometry. AB - BACKGROUND: Determination of mitochondrial membrane potential (DeltaPsim) is widely used to characterize cellular metabolism, viability, and apoptosis. Changes of DeltaPsim induced by inhibitors of oxidative phosphorylation characterize respective contributions of mitochondria and glycolysis to adenosine triphosphate (ATP) synthesis. METHODS: DeltaPsim in BSC-40 and HeLa G cell lines was determined by flow cytometry and spectrofluorometry. Its changes induced by specific mitochondrial inhibitors were evaluated using 3,3' dihexyloxacarbocyanine iodide (DiOC6(3)), tetramethylrhodamine ethyl ester, and MitoTracker Red. Mitochondrial function was further characterized by oxygen consumption. RESULTS: Inhibition of respiration by antimycin A or uncoupling of mitochondria by FCCP decreased DeltaPsim in both cell lines. Inhibition of ATP production by oligomycin or atractyloside induced a moderate decrease of DeltaPsim in HeLa G cells and an increase of DeltaPsim in BSC-40 cells. Statistically significant differences in DeltaPsim between the two cell lines were found with both flow cytometry and spectrofluorometry. Respirometry showed higher basal and FCCP-stimulated respiration in BSC-40 cells. CONCLUSION: Changes of DeltaPsim and oxygen consumption showed that BSC-40 cells are more sensitive than HeLa G cells to inhibitors of mitochondrial function, suggesting that BSC-40 cells are more dependent than HeLa G cells on aerobic ATP production. Determination of DeltaPsim changes by flow cytometry exhibited greater sensitivity than the ones by spectrofluorometry. PMID- 12655655 TI - Leukocyte apoptosis in whole blood involves platelet-dependent coaggregation. AB - BACKGROUND: Activated leukocytes and platelet-leukocyte interaction are involved in the pathogenesis of thrombotic and inflammatory events. Because apoptosis is a prerequisite for the successful resolution of an inflammatory response, we investigated the amount of apoptotic peripheral blood leukocytes (PBLs) in whole blood and their possible functional relation with the platelet-leukocyte interaction by a flow cytometric assay using APO 2.7 antibody for the detection of apoptosis METHODS: Thirty healthy subjects volunteered for the study. PBL apoptosis in seven volunteers was induced by phorbol 12-myristate 13-acetate or while standing at rest. RESULTS: Apoptosis was observed in all types of leukocytes (0.7% neutrophils, 1.5% monocytes, and 0.3% lymphocytes). Apoptosis was found predominantly in platelet and leukocyte coaggregates (<1% of nonaggregated leukocytes vs. 9% of platelet and leukocyte coaggregates). This phenomenon was even more pronounced after induction of leukocyte apoptosis in vitro (66% of platelet and leukocyte coaggregates). CONCLUSIONS: Apoptosis and platelet-leukocyte interaction seemed to be closely related phenomena, and apoptotic leukocytes seemed to trigger adhesion and, hence, activation of platelets. Because platelet-leukocyte interaction is involved in the pathogenesis of thrombotic events, apoptotic leukocytes may constitute an additional prothrombotic trigger. PMID- 12655656 TI - Measuring containment of viable infectious cell sorting in high-velocity cell sorters. AB - BACKGROUND: With the advent of high-speed sorters, aerosols are a considerable safety concern when sorting viable infectious materials. We describe a four-part safety procedure for validating the containment. METHODS: This procedure includes aerosol containment, physical barriers, environmental controls, and personal protection. The Aerosol Management System (AMS) produces a negative pressure within the sort chamber, where aerosols are forced through a HEPA filter. Physical barriers include the manufacturer's standard plastic shield and panels. The flow cytometer was contained within a BSL-3 laboratory for maximum environmental control, and the operator was protected by a respiratory system. Containment was measured by using highly fluorescent Glo-Germ particles under the same conditions as the cell sort. RESULTS: Escaping aerosols were vacuumed for 10 min onto a glass slide and examined. With the AMS active and the cytometer producing the maximum aerosols possible, Glo-Germ particles remained within the sort chamber. Measurements taken directly outside the door averaged fewer than one particle per slide, and those taken at 2 ft away and on top of the sorter were completely negative. CONCLUSIONS: With this monitoring system in place, aerosols can be efficiently measured, thus reducing the risk to the operator while sorting viable infectious cells. PMID- 12655657 TI - Perspectives. Faster, stronger, better: can state prescription aid fill the Medicare gap? PMID- 12655658 TI - Perspectives. Stretching Social Security retirement age hits Medicare and private pension plans. PMID- 12655659 TI - [Overview and expectation for the 21st century]. PMID- 12655661 TI - [Prospect for the 21st century: the circulatory organs]. PMID- 12655660 TI - [Prospect for the 21st century. The digestive organs]. PMID- 12655662 TI - [Prospect for the 21st century: endocrine metabolism]. PMID- 12655663 TI - [Prospect for the 21st century: the kidney]. PMID- 12655664 TI - [Prospect for the 21st century: the respiratory organs]. PMID- 12655665 TI - [Prospect for the 21st century: the blood]. PMID- 12655666 TI - [Prospect for the 21st century: neurology]. PMID- 12655667 TI - [Prospect for the 21st century: allergic collagen diseases]. PMID- 12655668 TI - [Prospect for the 21st century: infections]. PMID- 12655669 TI - [Genomic medicine]. PMID- 12655670 TI - [Translational research]. PMID- 12655673 TI - [Medical engineering]. PMID- 12655672 TI - [Cell transplantation and tissue engineering]. PMID- 12655671 TI - [Regeneration medicine]. PMID- 12655674 TI - [Medical application of nanobiotechnology--new development in gene diagnosis and gene therapy]. PMID- 12655675 TI - [New approach in drug designing: molecular-targeted drugs]. PMID- 12655676 TI - [Telemedicine]. PMID- 12655677 TI - [EMB (evidence-based medicine) and quality of health care]. PMID- 12655679 TI - [Information disclosure in medicine and its impact]. PMID- 12655678 TI - [Aging society and the nursing care insurance system--a report by a clinic in charge of regional health care]. PMID- 12655680 TI - [Specialist system in Japan--relaxation of the regulations concerning specialist announcement and the responsibility of medical societies]. PMID- 12655681 TI - [Global warming and changes in disease patterns]. PMID- 12655682 TI - [Revision of the National Health Insurance System and the practice of internal medicine]. PMID- 12655684 TI - [Education in internal medicine and the core curriculum]. PMID- 12655683 TI - [Requirement for clinical internship and training in specialties related to internal medicine]. PMID- 12655685 TI - [Digitalization of clinical information and its application to related medical services]. PMID- 12655686 TI - [The role of the internist in community medicine]. PMID- 12655687 TI - [Environmental problems and internal medicine--from the experience with Minamata Disease]. PMID- 12655688 TI - [Expectation on the Japanese Society of Internal Medicine]. PMID- 12655689 TI - [Expectation on the Japanese Society of Internal Medicine]. PMID- 12655691 TI - [Expectation on the Japanese Society of Internal Medicine]. PMID- 12655690 TI - [Expectation on the Japanese Society of Internal Medicine]. PMID- 12655692 TI - [Expectation on the Japanese Society of Internal Medicine--examination of the "ideal" and the "basic policy"]. PMID- 12655693 TI - [Swallow syncope]. PMID- 12655694 TI - [Tetanus with facial palsy as a unique initial symptom]. PMID- 12655695 TI - [Three times high-dose chemotherapy and peripheral blood stem cell transplantation rescue for patient with refractory ovarian germ cell tumor]. PMID- 12655697 TI - [Autoimmune hepatitis, Basedow's disease and myasthenia gravis]. PMID- 12655696 TI - [Genetic analysis of a case of Gitelman syndrome]. PMID- 12655698 TI - [Wegener's granulomatosis with pachymeningitis]. PMID- 12655699 TI - [Difficulties in the treatment of tuberculosis in infants]. AB - Nowadays, methods of the diagnosis in infants with suspected tuberculosis and of the treatment are definitely established in Japan, where a number of childhood tuberculosis has been falling down (the incidence is less than 2 per 100,000). Still, infants less than one year are considered to be at high risk against tuberculosis. Actually, the number of tuberculosis among them is three times larger than those of one or two years old children. One of major reasons of difficulties in the treatment is the rapid progress of the disease because of underdeveloped cell-mediated immunity among them. Alveolar macrophage and lymphocyte and their cooperation in immunological functions do not develop enough to kill or confine Mycobacterium tuberculosis. As a result, the infection may progress to disease quickly, and then may spread systemically before the starting treatment. Anatomical underdevelopment of cranial arteries and narrow cerebrospinal passages easily cause cerebral infarction and hydrocephalus following meningeal inflammation due to tuberculosis. These neurological disorders may result in poor prognosis despite of administration of effective anti-tuberculosis medicines. Delay in the diagnosis also makes the treatment difficult in some infants whose tuberculin skin test shows false negative and radiographic manifestation of chest is not clear. During the treatment, systemic and enteric viral infections occur frequently among infants with tuberculosis, and liver functional disorders caused by these infections sometimes disturbs the treatment for tuberculosis. Recurrence of tuberculosis is very rare among infants who complete the full treatment at the age of more than one year. Finally, it is important for the early start of treatment for tuberculosis to recognize both susceptibility to tuberculosis and difficulties in the diagnosis in some infants less than one year. PMID- 12655700 TI - [The incidence rate of active pulmonary tuberculosis among adult population with fibrotic lesions]. AB - The incidence rate of "active" pulmonary tuberculosis (TB) cases with bacteriological confirmation or cavitary lesions on chest radiographs was studied among population screened with mass miniature radiography in Funai-Gun, Kyoto Prefecture from 1982 to 1993. The results were as follows: Among population of 40 and over, prevalence rate of all fibrotic lesions on chest radiographs among male (8.3%) was twice as high as that of female (3.8%). The rate of moderate or extensive fibrotic lesions among male (3.3%) was three times as high as that of female (1%). The higher the age of the population, the higher the prevalence rate of radiological fibrotic lesions both in male and female. In male, in particular, prevalence rate of moderate or extensive fibrotic lesions started to rise after 40 years of age, became much higher after 70 years of age and reached 8.1% after 80 years of age. In female, however, it started to rise at 50s (0.3%) gradually and reached only 2.3% after 80 years of age. The incidence rate of "active" pulmonary TB among male of 40 years and over with moderate or extensive fibrotic lesions (4.2 per 1000 person-years) was 16 times as high as male of 40 years and over with normal chest X-ray finding (0.26 per 1000 person-years). Similarly, the incidence rate of "active" pulmonary TB in female of 40 years and over with moderate or extensive fibrotic lesions was 24 times as high as among female with normal finding, and the difference was statistically significant (p < 0.001). From the data obtained and bibliographical review, benefits of INH prophylaxis were discussed. PMID- 12655701 TI - [Effect of glycyrrhizin on anti-tuberculosis drug-induced hepatitis]. AB - In cases in which hepatotoxicity developed during anti-tuberculosis chemotherapy, the rapid recovery of liver function is essential for the completion of the anti tuberculosis chemotherapy protocol. Glycyrrhizin (Stronger Neo-Minophagen C: SNMC) is widely used in Japan for the treatment of patients with drug eruption or chronic hepatitis. However, a consensus on the clinical effects of glycyrrhizin for the treatment of anti-tuberculosis drug-induced hepatitis has not yet been reached. We studied 24 cases who showed abnormal liver function test results while undergoing anti-tuberculosis chemotherapy and who were treated with or without glycyrrhizin. We then compared recovery periods of liver function among both groups. The time required for liver function normalization in the patients who received glycyrrhizin (SNMC, 40 ml daily, intravenously) was 15.1 +/- 4.5 days and the time required for normalization in the non-glycyrrhizin group was 15.2 +/- 5.2 days. The difference was not significant and the fact indicated that glycyrrhizin is not useful for the treatment of anti-tuberculosis drug-induced hepatitis. PMID- 12655702 TI - [Notification rate of TB in Japan (1949-2001)]. AB - Notification of TB cases was introduced since 1947 in Japan, and statistics on notified TB cases by sex and age groups was available since 1949. Sex and age specific TB notification rates per 100,000 from 1949 to 2001 were tabulated together with the number of notified TB cases since 1969. PMID- 12655703 TI - [A case of pulmonary tuberculosis initially presented with syndrome of inappropriate secretion of antidiuretic hormone (SIADH)]. AB - A 90-year old man was admitted to a hospital because of consciousness loss with hyponatremia. Although his symptom promptly improved with Na supply, his chest X ray film showed pulmonary infiltration and direct microscopy of sputum smear was positive for acid-fast bacilli, then he was referred our hospital and was admitted. We made a clinical diagnosis of pulmonary tuberculosis with SIADH based on detailed examinations. But he should neither respiratory symptoms nor fever. He was medicated with the standard antituberculosis drugs with fluid restriction, and his tuberculosis and hyponatremia were improved gradually. We should be more careful about pulmonary tuberculosis irrespective of its severity as a cause of SIADH. PMID- 12655704 TI - [Effects of antisense oligo DNA on the antimicrobial activity of reactive oxygen intermediates and antimycobacterial agents against Mycobacterium avium complex]. AB - There has not yet been systematic studies which attempt to elucidate detailed profiles of the interaction between antimicrobial drugs and macrophage microbicidal mechanisms. We examined the effects of antisense oligo DNAs (AsDNAs) against oxyR and ahpC on the susceptibility of Mycobacterium avium complex (MAC) to the H2O2-halogenation system and combined antimycobacterial drugs [clarithromycin (CAM) + rifampicin (RFP)], both separately and in combination. It was found that AsDNA treatment of MAC did not affect the susceptibility of the organisms to any of the antimicrobial systems tested. Since the present AsDNAs did not efficiently reduce the expression of AhpC mRNA, attempts to increase bacterial uptake of AsDNAs are necessary to achieve significant increase in the drug susceptibility of MAC organisms due to AsDNA treatment. PMID- 12655705 TI - [Tuberculosis control strategy in Asia for the 21st century--tuberculosis control in the Western Pacific Region of World Health Organization]. AB - The unprecedented and rapid changes at the global level posed a big challenge to public health. The tuberculosis disease as one of major health problems today should be viewed from this context. These global challenges include 1) population issue particularly on growth and ageing; 2) epidemiological issue such as health transition; and, 3) social and environmental issues such as rapid urbanization and global warming. Furthermore, we should also take into account other changes such as the role of the government in the health service delivery, clinical or cure-oriented approach to prevention, increasing consumer demand and health financing. The burden of tuberculosis is devastating. Everyday in the Western Pacific Region (WPR), about 1000 people lose their life due to tuberculosis. Most of them are between the ages 14-54, which are the so-called economically productive age group. The economic impact, therefore, is significant. In addition TB is a disease of poverty of which the risk of getting TB is higher in poor who have less access to TB services due to financial barrier and lack of knowledge. Despite this devastating situation, TB has a significant and cost effective tool called DOTS. WPRO put highest priority in TB control programme not only because of the facts mentioned above but I also consider TB as an agenda carried over from the last century. I would like, then, to commit myself to this cause for the betterment of the future of the next generation. PMID- 12655706 TI - [HIV infection and tuberculosis]. AB - The number of people infected with human immunodeficiency virus (HIV) is gradually increasing in Japan, and the morbidity rate from tuberculosis in the Japanese people is high. Accordingly, the number of cases with both infections is considered to increase in the future. Our hospital has already encountered 31 cases of HIV associated tuberculosis. HIV infects mainly CD4-positive cells. The extreme decrease in the cell count results in serious cellular immunological disorder. CD4-positive cell disorder induces disorders of B lymphocytes, cytotoxic T cells, natural killer cells, and macrophage functions. These destructive conditions show the state of immunodeficiency including macrophage that are most important for defense of acid-fast bacterial infection. Migration and activation of macrophages with cytokines derived from T cells are impaired to induce the following phenomena: hypoplasia of granuloma, failure of tubercule bacillus suppression, the spread to regional lymph nodes (hilar or mediastinal lymph nodes), and hematogenous dissemination. On this occasion, caseous necrosis and cavitation are unlikely to occur, and false-negative tuberculin reaction is often observed. The incidence of severe cases, which include miliary tuberculosis, tuberculous meningitis, etc., and extrapulmonary tuberculosis, are high among acquired immunodeficiency syndrome (AIDS)-associated tuberculosis cases. HIV-infected tuberculosis cases are generally regarded as endogenous exacerbation, but they include primary infection and reinfection as well. Even during the treatment for drug-sensitive strains particularly, some cases may have reinfection with multidrug-resistant bacteria, suggesting that caution should be taken against this point. Conversely, the association of tuberculosis is a factor for the poor prognosis of HIV infection, since it facilitates the development of HIV infection. If the bacteria belong to a drug-sensitive strain, the infection with them responds well to antituberculous drugs, the same as in tuberculosis cases without HIV infection, showing a favorable prognosis. However, the mortality rate of infection with multi-drug-resistant tuberculosis is extremely high. The combined use of a protease inhibitor, i.e., anti-HIV drug, with rifampicin is regarded as contraindication for the treatment because rifampicin strongly induces hepatic cytochrome P-450 and increases the metabolism of protease inhibitors and nonnucleoside reverse transcriptases to markedly decrease the blood concentrations. Accordingly, the treatment for tuberculosis should take priority over that for HIV infection in HIV-infected tuberculosis, and highly active antiretroviral therapy (HAART) may be administered after the treatment of tuberculosis. When HAART is necessary for the treatment during the tuberculosis treatment, rifampicin had better be exchanged to rifabutin because the effect of rifabutin to induce cytochrome P-450 is less potent than that of rifampicin. A report has recently shown that the exacerbation of pyrexia and chest X-ray findings was transiently observed approximately 2 weeks after potent anti-HIV therapy for HIV-infected tuberculosis, which included a protease inhibitor. The reason for the exacerbation has been believed to be that the impaired function of CD4-positive cells is improved by the administration of anti-HIV drugs to raise temporarily the reaction of the vital part to M. tuberculosis. A tuberculin skin test (TST) reaction size of > or = 5 mm of induration is considered positive (i.e., indicative of M. tuberculosis infection) in persons who are infected with HIV. Persons with a TST reaction size > or = 5 mm who have not previously received treatment for M. tuberculosis infection should receive tuberculosis preventive treatment. Prevention by BCG vaccination is regarded as contraindications for HIV-infected patients, because disseminated M. bovis infection may be associated with them. Many HIV-positive patients infected with tuberculosis show uneventful healing, when M. tuberculosis is the sensitive strain. However, since some patients show the rapid course of tuberculosis, clinical physicians keep the early detection of tuberculosis for HIV-infected patients and the association of HIV infection for tuberculosis patients in their mind, respectively. PMID- 12655707 TI - [Method of embryological engineering to elucidate the influence of genetic and maternal factors on craniofacial development in mice]. PMID- 12655709 TI - [Message to junior anesthetists]. PMID- 12655708 TI - A Relationship between febrile illness, serum albumin level and mortality in elderly hospitalized patients. AB - The author sought to determine the etiology of febrile illness and risk factors influencing mortality in elderly patients. The patients, aged 65 years or older, were hospitalized in a geriatric hospital in Fukuoka City, Japan. A total of 405 febrile illness was diagnosed and the background, clinical features of the fever, and laboratory data were analyzed. Serum albumin levels were checked both at the onset and on the 8th day of fever in 140 patients. Of febrile illness of elderly patients, pneumonia is a principal disease with a high risk of mortality. Serum albumin level at the onset of fever and the reduction rate in the serum albumin level from baseline to day +8 after the onset of febrile illness were predictive factors of mortality within 30 days of fever. When examining febrile, elderly patients, checking the serum albumin level is useful for determining which patients are at high risk. PMID- 12655710 TI - [Post-genome sequence era and medicine]. PMID- 12655711 TI - [Oxidative stress protection by heme]. PMID- 12655712 TI - [Progress in study of erythrocytes]. PMID- 12655714 TI - [Progress in cardiovascular surgery]. PMID- 12655713 TI - [Regulative mechanism in smooth muscle tonus]. PMID- 12655716 TI - [Etiological view of diseases from protein domain--activation mechanism of reactive-oxygen-species producing phagocyte NADPH oxidase important rule in host defence]. PMID- 12655715 TI - [Mechanism of macrophage activation by LPS]. PMID- 12655717 TI - [Regulative mechanism of calcium signaling in cardiomyocytes]. PMID- 12655718 TI - [Functional analysis of pain pathway in posterior horn cells using spinal cord slice and in-vivo patch clamp method]. PMID- 12655719 TI - [Recent progress in therapy for postherpetic neuralgia]. PMID- 12655720 TI - [Fetal monitoring]. PMID- 12655721 TI - [Trends in therapy for diabetes mellitus]. PMID- 12655722 TI - [Physiological study on nociception]. PMID- 12655723 TI - [Recent progress in the field of pediatric surgery]. PMID- 12655724 TI - [Oxygen metabolism in acute phase reaction]. PMID- 12655725 TI - [Organ-protective effects of anesthetics]. PMID- 12655726 TI - [Basic and clinical studies on coronary vasospasm]. PMID- 12655727 TI - [Future of hematological intensive care in the next 15 years]. PMID- 12655728 TI - [Neuroscience and anesthesia]. PMID- 12655729 TI - [Recent study on action mechanism of general anesthetics]. PMID- 12655730 TI - [Education for anesthetists]. PMID- 12655731 TI - [Clinical skill and knowledge of specialists in respiratory management]. PMID- 12655732 TI - [Next generation of drugs used in anesthesia]. PMID- 12655733 TI - [Update on pediatric anesthesia]. PMID- 12655734 TI - AARN applauds Romanow Report. Urges all levels of government to work together to improve health care. PMID- 12655735 TI - Alberta RN statistics 2002. Registered nursing by the numbers. PMID- 12655736 TI - R.E.L.A.T.E. and R.E.S.P.O.N.D. Key tools for building positive relationships with patients and families. PMID- 12655737 TI - The upper part of the earnings distribution in the United States: how has it changed? AB - This article uses Social Security Administration data to examine changes in the upper end of the earnings distribution over the period 1982-1995. These data provide a unique opportunity for analyzing those changes because they come directly from W-2 forms and are not topcoded--obvious advantages over data typically provided by surveys such as the Current Population Survey. Although they do not provide some of the information necessary to explain what one observes occurring at the top of the earnings distribution (no educational attainment information, for example), these data are sufficient for describing in great detail what happened to high earners through the 1980s and into the 1990s. This analysis clearly demonstrates the extent to which earnings are concentrated at the top of the distribution. The study's findings reinforce those of Feenberg and Poterba (2000) by showing that the very highest earners--those in the upper 1 percent--experienced the largest relative increase in earnings share from 1982 to 1995. Even within that upper 1 percent, those with earnings in the upper 0.1 percent were the ones driving the increase in the group's earnings share. Perhaps not surprisingly, the overwhelming majority of the highest earners are white men who are in the middle to latter part of their working lives. Women have made strides toward entering this elite group of earners but still form a very small percentage of the group relative to their size in the working population. Very few blacks are in the extreme upper tail of the earnings distribution, and they have made very little progress (in absolute terms) over the period 1982-1995 in increasing their numbers. In contrast, persons of racial/ethnic backgrounds other than white or black have increased their presence among top earners. They went from being relatively underrepresented in the top 0.1 percent in 1982 to being overrepresented by 1995; that is, they accounted for a larger share of the top 0.1 percent of earners than they did of the entire working population. Finally, the study also finds that the percentage of overall wage and salary earnings from Social Security-covered employment that is not taxed for the purposes of Old-Age, Survivors, and Disability Insurance because of the taxable maximum generally increased over the 1982-1995 period. Wage and salary earnings above the taxable maximum rose at a faster rate than is the average wage. PMID- 12655738 TI - Trends in the economic status of the elderly, 1976-2000. AB - The economic well-being of elderly Americans (aged 65 or older) improved between 1976 and 2000. Overall, poverty rates fell during this period, median real income rose, and median income relative to the working-age population was relatively stable. Most population subgroups shared in the reduced poverty rates; however, the economic status of elderly Hispanics did not improve. This article attempts to explain those economic trends by identifying changes in five sources of income for the elderly and analyzing the changes in the context of demographic changes in the elderly populations over the past 25 years. As a result of increased longevity, for example, larger proportions of elderly men and women are now 80 or older, and smaller proportions are 65 to 69. Hispanics and Asian Americans make up a larger share of the elderly population and whites a smaller share. The fraction of women who are married has increased, the fraction who are widowed has fallen, and the fraction who are divorced has grown. Such demographic changes can greatly affect the economic status of subgroups as well as the overall elderly population. Of the five sources of income for the elderly, Social Security remains the most prevalent and important. While both the rate of receipt and the share of aggregate income from Social Security benefits stayed relatively steady over the past 25 years, the average real Social Security benefit increased because of rising wages. Income from assets, the second most important source of income for the elderly, fluctuated. Because the elderly are more likely to hold interest-bearing assets such as bonds rather than stocks, their asset income is responsive to changes in nominal interest rates and bond yields. Receipt of pension income increased during this period, although it leveled off during the 1990s. Factors contributing to this pattern include enactment of the Employee Retirement Income Security Act of 1974, which increased protections of pension benefits for spouses, and improved labor market opportunities for blacks and women. In recent years, defined contribution pension plans have become more prevalent than defined benefit plans, but the full effect of this change on pension income may not yet be apparent. After decades of decline, labor force participation rates of older men leveled out in the mid-1980s and then increased. For older women, the trend before the mid-1980s was flat, but since then rates have risen substantially. The increased use of part-time jobs or self-employment to ease the transition into retirement, the economic expansion of the 1990s, and the liberalization of the Social Security earnings test may all have contributed to those trends. Although the percentage of elderly people with earnings has increased only modestly in the past few years, the share of income from earnings has grown substantially--from 16 percent of income in 1984 to 23 percent in 2000. Finally, Supplemental Security Income (SSI) benefits are indexed for inflation but not for growth in real wages. As real incomes of the elderly rose, therefore, fewer elderly persons were eligible to receive SSI or, for those receiving SSI, were eligible for smaller benefits. The proportion of elderly persons receiving public assistance, primarily SSI, declined from 11 percent in 1976 to 5 percent in 2000. PMID- 12655739 TI - Income growth and future poverty rates of the aged. AB - This article estimates the effects of 50 years of steady growth in incomes on poverty rates among the elderly. It assumes that the poverty threshold continues to be adjusted for inflation but not for increases in real incomes. Simulations with the March 1998 Current Population Survey indicate that if the benefit rules for Social Security and Supplemental Security Income (SSI) are not changed and if earnings and other sources of income in an otherwise unchanging population grow by 1 percent per year (the intermediate assumption about earnings growth used in the Social Security Trustees' Report), poverty among the elderly will decrease from 10.5 percent in 1997 to about 7.2 percent in 2020 and to 4.1 percent in 2047. These projected poverty rates are quite sensitive to both the assumption about earnings growth and the assumption that benefits are not further reduced to maintain solvency. This article quantifies the sensitivity of the results to these assumptions and discusses several other aspects that might affect future poverty rates--changes in other income components like SSI, earnings, and pensions; changes in longevity and marital patterns; and changes in the distribution of earnings. PMID- 12655740 TI - SSI eligibility and participation among the oldest old: evidence from the AHEAD. AB - The proportion of elderly SSI recipients aged 70 or older has been growing in recent years, perhaps because of rising life expectancies overall and a higher incidence of poverty among the oldest old. In 1999, 84 percent of all elderly SSI recipients were 70 or older. This article examines Supplemental Security Income (SSI) eligibility and participation among the oldest old. The analysis was based on 1993 data from the Study of Assets and Health Dynamics Among the Oldest Old that were used to build a detailed SSI eligibility model to identify individuals who meet the federal criteria for SSI income and resource eligibility. The participation rate among those eligible for federal SSI benefits is 53.9 percent, which is generally consistent with the findings of other studies. Furthermore, eligible participants would receive a significantly higher federal SSI benefit than eligible nonparticipants. Correspondingly, eligible participants have significantly lower incomes and assets than eligible nonparticipants. An econometric model is used to estimate the influence of various demographic, financial, and health care use characteristics on the probability of SSI participation among eligible individuals and couples. The model corrects for measurement error in calculated benefits and for misclassifying someone as ineligible. The empirical results show that the effect of higher SSI benefits on the probability of participation is substantial--a $100 increase in benefits would increase the probability of participating for an average eligible unit by 15 percentage points. Many of the demographic, financial, and health care use variables also are important predictors of SSI participation among the oldest old. The eligibility and participation models are also used to simulate the effect of increasing the SSI unearned income disregard from $20 to $125. Those made eligible by this policy change would receive a very low federal SSI benefit on average, suggesting that they are on the margin of eligibility under the original program rules. The simulated participation rate is 48.8 percent--5 percentage points lower than under the original program rules--reflecting the low benefit that new eligibles would receive. Only 36 percent of those made eligible by the new program rules are predicted to participate. These SSI eligibility and participation models are potentially useful tools for policy analysis. It is fairly straightforward to use these models to change a feature of SSI eligibility, reestimate the group of eligible individuals and couples, and predict participation among those who are eligible under the simulated program rules. New eligibles can be compared with those eligible under original program rules. New participants can be compared with old participants. Although these models focus only on individuals aged 70 or older, this type of analysis can be helpful in estimating the potential distributional effects of proposed SSI policy changes. PMID- 12655741 TI - What determines 401(k) participation and contributions? PMID- 12655743 TI - Will foundation trusts wield the axe on nursing posts? PMID- 12655742 TI - Comparing beneficiaries of the Medicare savings programs with eligible nonparticipants. AB - This note focuses on participation in two entitlement programs that help reduce out-of-pocket expenses for low-income Medicare beneficiaries: the Qualified Medicare Beneficiary (QMB) program and the Specified Low-Income Medicare Beneficiary (SLMB) program. As of 1999, about 2.75 million eligible, noninstitutionalized individuals were not enrolled in these Medicare savings programs. The eligible nonparticipants differed substantially from the QMB and SLMB participants in that they were less likely to be Supplemental Security Income beneficiaries and more likely to be elderly, nonblack, and in relatively good health. These findings, which could help target future outreach efforts, are based on Survey of Income and Program Participation data matched with administrative records from the Social Security Administration. PMID- 12655744 TI - Should clinical placement time be decreased? PMID- 12655745 TI - Nursing the nurses. PMID- 12655746 TI - The use of protective isolation. AB - Protective isolation denotes a range of practices used to protect immunocompromised hospital patients from infection. The decision to institute protective isolation will be made as part of an individualized plan of care, taking into account the reason for, the nature, degree and expected duration of immunosuppression of the affected person. This article examines protective isolation, when it might be implemented and the evidence for the use of single rooms. The issues of hand hygiene, food, drink and equipment are also discussed. PMID- 12655747 TI - What you need to know about... haemorrhoids. PMID- 12655748 TI - The use of nebulisers. PMID- 12655749 TI - The ethics of assisted suicide. AB - There is continuing national and international debate with reference to autonomy and the right to choose the manner and time of our death. It is now recognised that human life has a natural end and that, as such, it should not be preserved simply because a treatment exists. Three cases are briefly discussed which raise the issue of whether or not we should be permitted by law to choose how and when to die. The ethical principle of autonomy and the influence of religion are considered. Central to the debate about end-of-life choices is the recognition that not all persons can be healed and not all suffering can be relieved. PMID- 12655750 TI - The medical nurse practitioner's role in early stroke recognition. AB - Stroke is one of the world's major health care problems and begins as an acute medical emergency. To ensure the best outcomes, stroke services should comprise an identified stroke unit with a dedicated multidisciplinary team of specialist staff working within agreed protocols. The medical nurse practitioner's intervention at the assessment stage for patients admitted to A&E with stroke should provide a more timely and appropriate referral in order to access stroke beds directly. PMID- 12655751 TI - What a difference a matron can make. PMID- 12655752 TI - Swapping a suit for a uniform. PMID- 12655753 TI - Understanding scabies. AB - Increased incidence of scabies in older people has highlighted difficulties of diagnosis and treatment. Ian Burgess describes the life cycle of the scabies mite, transmission of the infection, diagnosis and treatment. PMID- 12655754 TI - Improving hand hygiene compliance. AB - Achieving compliance with hand hygiene can be difficult. Annette Jeanes explains how a new approach to marketing hand hygiene produced sustained positive results. PMID- 12655756 TI - The psychological impact of isolation. AB - Maurice Madeo describes the psychological effect of source isolation and strategies for dealing with the effects on the patient. PMID- 12655755 TI - The delivery of hepatitis B vaccine. AB - Hepatitis B is a serious preventable infection that is common in men who have sex with men. Historically, uptake of the vaccine has been poor in this group. Leigh Chislett describes how the introduction of a nurse-led service significantly increased uptake of the vaccine at a London clinic. PMID- 12655757 TI - A nurse's experience of isolation. AB - A nurse specialist in infection control, describes how it felt to be on the receiving end of protective isolation. PMID- 12655759 TI - Listening online. PMID- 12655760 TI - It's all in the mix. PMID- 12655761 TI - What do you think of the deal so far? PMID- 12655763 TI - Fertile minds. PMID- 12655762 TI - Doctor's orders. PMID- 12655764 TI - Pilot evaluation of an intermediate care scheme. AB - AIM: To explore whether an intermediate care scheme could reduce the number of unnecessary emergency hospital admissions for people aged over 65. METHOD: A retrospective documentary analysis of 57 patients' discharge summaries was undertaken. RESULTS: The analysis identified that only 7 per cent (n = 4) of these patients were re-admitted to hospital. These admissions appeared to be necessary in light of increasing clinical instability. Thirty three per cent (n = 19) of the patients required no further interventions to remain at home. Forty seven per cent (n = 27) of the patients that remained at home were referred to other agencies for ongoing care. The most common need identified was meeting patients' hygiene needs. CONCLUSION: This pilot evaluation identified that only 5 per cent of patients were re-admitted to hospital, which demonstrated great potential. The integration of a health and social care team appears to have resulted in a service with the skill, vision and resources to respond to older people's needs, ensuring that they are cared for, where appropriate, at home, with the right support. PMID- 12655765 TI - Setting up a deep vein thrombosis clinic. AB - The author describes the establishment of an outpatient service for patients with a suspected deep vein thrombosis. The outpatient service is on a medical admission unit at Glenfield Hospital, one of three sites of the University Hospitals of Leicester. The author considers that the service has been as effective as the inpatient service previously provided and in its first year of operation, 400 potential bed days were saved on positive to DVT patients. PMID- 12655766 TI - The new Code of Professional Conduct. AB - The Nursing and Midwifery Council (NMC) has approved a new Code of Professional Conduct (NMC 2002a). This article discusses the main elements of the new code, examines the implications for the profession and encourages you to think about the implications for your own nursing practice. It identifies actions that you should take to comply with the code. PMID- 12655767 TI - Nutritional support in wound healing. AB - Wounds place increased metabolic and hence nutritional demands on patients, which may not always be met, especially in older patients in the community. Patients may be deficient in carbohydrates, protein and amino acids, fatty acids and certain micronutrients. Nutritional support may be needed to promote wound healing. Nurses should be able to assess the risk of poor nutrition in patients with wounds and have a role in ensuring adequate nutrition through the normal diet or supplementation where necessary. PMID- 12655768 TI - Foot care in patients with diabetes. AB - This article outlines the three main factors that cause foot problems in people with diabetes: peripheral neuropathy, peripheral vascular disease and infection. It describes how these problems are classified and how nurses can use their skills in assessment and education to improve outcomes for these patients. PMID- 12655769 TI - Legal concerns in tissue viability and wound healing. AB - In an age of increasing litigation, it is important for tissue viability practitioners to be aware of the legal issues surrounding their practice. Bridgit Dimond discusses some of the legal aspects that were raised as concerns by participants recently attending a post-graduate diploma course in tissue viability and wound healing. She clarifies the main legal requirements for healthcare professionals working in this field. PMID- 12655770 TI - Functional post-translational proteomics approach to study the role of N-glycans in the development of Caenorhabditis elegans. AB - Glycosylation is one of the most common post-translational protein modifications. Carbohydrate-mediated interactions between cells and their environment are important in differentiation, embryogenesis, inflammation, cancer and metastasis and other processes. Humans and mice with mutations that prevent normal N glycosylation show multi-systemic defects in embryogenesis, thereby proving that these molecules are essential for normal development; however, a large number of proteins undergo defective glycosylation in these human and mouse mutants, and it is therefore difficult to determine the precise molecular roles of specific N glycans on individual proteins. We describe here a 'functional post-translational proteomics' approach that is designed to determine the role of N-glycans on individual glycoproteins in the development of Caenorhabditis elegans. PMID- 12655771 TI - Comparative aspects of glycosyltransferases. AB - Glycosyltransferases, the enzymes that build oligosaccharides and glycoconjugates, have received much interest in recent years owing to their biological functions and their potential uses in biotechnology. Despite the fact that many glycosyltransferases recognize similar donor or acceptor substrates, there is surprisingly limited sequence identity between different classes. On the one hand, the glycosyltransferases are found in a large number of families, by sequence-based classification. On the other hand, only two structural folds have been identified among the fewer than one dozen glycosyltransferases that have been crystallized at present. Detection of conserved motifs that have a direct role in the functional aspects of glycosyltransferases is one approach for identifying remote similarity. With the availability of more crystal structures, the use of the fold-recognition approach is also very promising. PMID- 12655772 TI - Glycosyltransferases and glycan structures contributing to the adhesive activities of L-, E- and P-selectin counter-receptors. AB - In mammals, leucocytes of the adaptive and innate immune systems must move from their sites of origin to sites of maturation, or to where they are deployed against the invasion of pathogens. The vascular tree serves as the primary throughfare by which leucocytes move to these various destinations. Adhesion must be established between the leucocyte and the endothelial cells that line the vascular tree to enable leucocytes to escape the vascular compartment and then contribute to extravascular immune processes. A major fraction of these leucocyte endothelial-cell adhesive events initiate with, and require interactions between, the selectin family of cell adhesion molecules and their glycoconjugate counter receptors. This article will review the structures of the glycan components of these counter-receptors, and the glycosyltransferases that control their expression. PMID- 12655773 TI - New insights into heparan sulphate biosynthesis from the study of mutant mice. AB - Heparan sulphate (HS) is an essential co-receptor for a number of growth factors, morphogens and adhesion proteins. The biosynthetic modifications involved in the generation of a mature HS chain may determine the strength and outcome of HS ligand interactions. These modifications are catalysed by a complex family of enzymes, some of which occur as multiple gene products. Various mutant mice have now been generated, which lack the function of isolated components of the HS biosynthetic pathway. In this discussion, we outline the key findings of these studies, and use them to put into context our own work concerning the structure of the HS generated by the Hs2st -/- mice. PMID- 12655774 TI - Genomic analysis of C-type lectins. AB - Many biological effects of complex carbohydrates are mediated by lectins that contain discrete carbohydrate-recognition domains. At least seven structurally distinct families of carbohydrate-recognition domains are found in lectins that are involved in intracellular trafficking, cell adhesion, cell-cell signalling, glycoprotein turnover and innate immunity. Genome-wide analysis of potential carbohydrate-binding domains is now possible. Two classes of intracellular lectins involved in glycoprotein trafficking are present in yeast, model invertebrates and vertebrates, and two other classes are present in vertebrates only. At the cell surface, calcium-dependent (C-type) lectins and galectins are found in model invertebrates and vertebrates, but not in yeast; immunoglobulin superfamily (I-type) lectins are only found in vertebrates. The evolutionary appearance of different classes of sugar-binding protein modules parallels a development towards more complex oligosaccharides that provide increased opportunities for specific recognition phenomena. An overall picture of the lectins present in humans can now be proposed. Based on our knowledge of the structures of several of the C-type carbohydrate-recognition domains, it is possible to suggest ligand-binding activity that may be associated with novel C type lectin-like domains identified in a systematic screen of the human genome. Further analysis of the sequences of proteins containing these domains can be used as a basis for proposing potential biological functions. PMID- 12655775 TI - Lectins and protein traffic early in the secretory pathway. AB - Lectins of the early secretory pathway are involved in selective transport of newly synthesized glycoproteins from the endoplasmic reticulum (ER) to the ER Golgi intermediate compartment (ERGIC). The most prominent cycling lectin is the mannose-binding type I membrane protein ERGIC-53 (ERGIC protein of 53 kDa), a marker for the ERGIC, which functions as a cargo receptor to facilitate export of an increasing number of glycoproteins with different characteristics from the ER. Two ERGIC-53-related proteins, VIP36 (vesicular integral membrane protein 36) and a novel ERGIC-53-like protein, ERGL, are also found in the early secretory pathway. ERGL may act as a regulator of ERGIC-53. Studies of ERGIC-53 continue to provide new insights into the organization and dynamics of the early secretory pathway. Analysis of the cycling of ERGIC-53 uncovered a complex interplay of trafficking signals and revealed novel cytoplasmic ER-export motifs that interact with COP-II coat proteins. These motifs are common to type I and polytopic membrane proteins including presenilin 1 and presenilin 2. The results support the notion that protein export from the ER is selective. PMID- 12655777 TI - Roles of galectins in vivo. AB - The mutational analysis of the galectin family is shedding a different light of this class of molecules. On the one hand, it appears that galectin 1 and galectin 3 are not required for the survival of mice in normal animal house conditions, while on the other hand, there seems to be several subtle, but very complex, consequences of lacking galectins during development. PMID- 12655776 TI - New I-type lectins of the CD 33-related siglec subgroup identified through genomics. AB - Siglecs are sialic-acid-binding proteins of the Ig superfamily that are involved in cell-cell interactions and signalling. In recent years, several novel siglecs that are highly related to CD33/Siglec-2 have been identified through genomics and functional screens. In addition to their distinct sialic-acid-binding properties, most of these novel siglecs bear tyrosine-based signalling motifs that are typically found in inhibitory receptors of the immune system. The restricted expression patterns of CD33-related siglecs in the haemopoietic and immune systems suggests that they are involved in regulating leucocyte activation during inflammatory and immune responses. PMID- 12655778 TI - MS screening strategies: investigating the glycomes of knockout and myodystrophic mice and leukodystrophic human brains. AB - The implementation of highly sensitive and rapid mass spectrometric screening strategies for defining the glycosylation repertoires of organs in knockout mice is helping to reveal the roles that glycans play in health and disease. Thus novel glycosylation pathways have been uncovered in two such knockouts, namely alpha-mannosidase II null mice and UDP-N-acetylglucosamine: alpha 6-D-mannoside beta 1,2-N-acetylglucosaminyltransferase II null mice. This chapter documents the glycosylation profiles of a wide range of organs from the normal mouse which should facilitate future glycomics studies of knockout mice. Furthermore, we report applications of our screening technology in studies of the myodystrophy mouse and a human leukodystrophy. PMID- 12655779 TI - The glycomes of Caenorhabditis elegans and other model organisms. AB - There is no doubt that the immense amount of information that is being generated by the initial sequencing and secondary interrogation of various genomes will change the face of glycobiological research. However, a major area of concern is that detailed structural knowledge of the ultimate products of genes that are identified as being involved in glycoconjugate biosynthesis is still limited. This is illustrated clearly by the nematode worm Caenorhabditis elegans, which was the first multicellular organism to have its entire genome sequenced. To date, only limited structural data on the glycosylated molecules of this organism have been reported. Our laboratory is addressing this problem by performing detailed MS structural characterization of the N-linked glycans of C. elegans; high-mannose structures dominate, with only minor amounts of complex-type structures. Novel, highly fucosylated truncated structures are also present which are difucosylated on the proximal N-acetylglucosamine of the chitobiose core as well as containing unusual Fuc alpha 1-2 Gal 1-2 Man as peripheral structures. The implications of these results in terms of the identification of ligands for genomically predicted lectins and potential glycosyltransferases are discussed in this chapter. Current knowledge on the glycomes of other model organisms such as Dictyostelium discoideum, Saccharomyces cerevisiae and Drosophila melanogaster is also discussed briefly. PMID- 12655780 TI - Custom microarray for glycobiologists: considerations for glycosyltransferase gene expression profiling. AB - The development of microarray technology offers the unprecedented possibility of studying the expression of thousands of genes in one experiment. Its exploitation in the glycobiology field will eventually allow the parallel investigation of the expression of many glycosyltransferases, which will ultimately lead to an understanding of the regulation of glycoconjugate synthesis. While numerous gene arrays are available on the market, e.g. the Affymetrix GeneChip arrays, glycosyltransferases are not adequately represented, which makes comprehensive surveys of their gene expression difficult. This chapter describes the main issues related to the establishment of a custom glycogenes array. PMID- 12655781 TI - Neoglycoconjugates: trade and art. AB - This chapter deals with the tendencies in design of multivalent neoglycoconjugates for glycobiology research and high-throughput profiling technologies, including cellular phenotyping. Soluble polyacrylamide (PAA) conjugates are remarkable owing to a variety of possibilities for synthesis and application. PAA is soluble and stable, and the molecule is flexible, PAA tethered ligands are capable of adjusting to a receptor during multiple-point interactions and PAA does not bind to the cell surface. Synthesis provides unlimited diversity of the probe types (biotin, fluorescein, allyl, digoxygenin, 3H, radiolabelled I), glyco-particles, glyco-surfaces, multiarrays, immunogens etc. Several examples illustrate the most advanced applications. (i) Dynamic systems: the selectin ligands immobilized on the surface as sugar-PAA conjugates made the study of the kinetics of rolling in the model system possible. Carbohydrate ligands that are covalently attached to the chip as sugar-PAA conjugates are of use with the surface plasmon resonance method. (ii) Pseudoglycoprotein: some questions arise regarding the biologically active glycoproteins, e.g.: is a carbohydrate or peptide fragment responsible for the activity? We have proposed the approach that promotes to answer this and other questions. The pool of oligosaccharides that were spitted off a glycoprotein is attached to PAA resulting in a pseudoglycoprotein. (iii) Virtual (dynamic) glycotope: receptor-ligand recognition, such as that of P-selectin with its receptor P-selectin glycoprotein ligand 1, frequently involves molecular interactions at two distinct sites. Using P-selectin as a model, we developed an approach to discover novel ligands. PAA was synthesized with multiple ligands; a marked synergistic inhibitory effect was observed. PMID- 12655782 TI - Fetal antigen 1 (FA1) in the adult rat adrenal gland, ovary and pituitary gland. AB - Fetal antigen 1 (FA1) is a circulating glycoprotein containing six epidermal growth factor (EGF)-like repeats. FA1's larger membrane-bound precursor is defined by the cDNAs referred to as either human delta-like (dlk) or human adrenal specific cDNA, pG2. In rodents FA1 has also been studied under the names of preadipocyte factor 1 (Pref-1), and zona glomerulosa-specific factor (ZOG). FA1 is abundantly expressed in fetal tissues, but in the mature cells of the adult organism the tissue presence of the protein seems to be restricted to neuroendocrine tissues. The present study demonstrates FA1 localisation in endocrine tissues of the adult female rat in which the protein was found present in the medulla and the zona glomerulosa of the cortex of the adrenal glands, in the pars distalis of the adenohypophysis, and in the ovarian granulosa lutein cells. No staining was found in the pancreas, which is in contrast to what has been described in the human. PMID- 12655783 TI - Increase in plasma fibrinogen and plasminogen activator inhibitor level in diabetic KK and KK-Ay mice. AB - Coagulation activity in KK mice and KK-Ay mice produced by transferring the yellow obese gene (Ay) into KK mice, was studied to examine whether both mice are useful as a model of diabetic atherosclerosis. Plasma levels of hemoglobin A1c (HbA1c), insulin, fibrinogen, plasminogen activator inhibitor (PAI) and thrombomodulin were significantly high in KK and KK-Ay mice compared with age matched non-diabetic mice (ddY mice). The changes in the plasma levels of fibrinogen at each time-point correlated with the increases in HbA1c levels. Pathological observation by Oil red O staining of aorta tissue from 4-month-old KK and KK-Ay mice revealed the early stages of atherosclerosis such as lipid deposition. These age-related increases in the plasma level of fibrinogen and PAI suggested that KK-Ay mice may contribute to help elucidate the early stages of diabetic atherosclerosis. PMID- 12655785 TI - Antitumor activity of doxorubicin in combination with docetaxel against human breast cancer xenografts. AB - In this study we assessed the in vivo antitumor activity of combined docetaxel (DOCE) and doxorubicin (DXR) treatment using 2 human breast carcinoma cell xenografts (R-27 and MX-1) in the nude mouse model. The transplanted tumors were allowed to reach exponential growth, whereupon 10 or 40 mg DOCE per kg alone (i.p.), 8 mg DXR per kg alone (i.v.), or 10 mg/kg DOCE (i.p.) and 8 mg/kg of DXR (i.v.), in the sequence of DOCE followed by DXR, were administered. The in vivo antitumor activity of combined DOCE and DXR was synergistic against R-27 and additive against MX-1. P-glycoprotein (P-gp) was detected immunohistochemically, and was highly expressed in R-27, but not in MX-1. In conclusion, DOCE may increase the antitumor activity of DXR against P-gp-positive breast cancer xenografts, such that the DOCE and DXR combination may be a useful treatment in clinical breast cancer. PMID- 12655784 TI - Manganese superoxide dismutase-plasmid/liposome (MnSOD-PL) intratracheal gene therapy reduction of irradiation-induced inflammatory cytokines does not protect orthotopic Lewis lung carcinomas. AB - BACKGROUND: Intratracheal injection of manganese superoxide dismutase plasmid/liposome (MnSOD-PL) prior to single fraction or fractionated irradiation of C57BL/6J mouse lung has been demonstrated to protect the lung from irradiation induced damage. MATERIALS AND METHODS: To determine whether irradiation-induced inflammatory cytokine levels influenced the recovery of tumors following single fraction irradiation, mice with orthotopic Lewis Lung Carcinoma (3LL) tumors received MnSOD-PL treatment 24 hours after tumor implantation and 24 hours prior to irradiation. Subgroups were implanted with Alzet pumps continuously replacing levels of inflammatory cytokines over 7 days. RESULTS: In cytokine-treated mice, there was no detectable significant alteration in radiotherapy-mediated improved survival (tumor regrowth delay) compared to irradiated control mice. Each group of mice that received MnSOD-PL had increased survival compared to irradiated controls. CONCLUSION: These results support the anticipated safety of intrapulmonary MnSOD-PL gene therapy in lung cancer patients for protection of normal lung tissue from irradiation while allowing effective irradiation-mediated tumor control. PMID- 12655786 TI - Human herpesvirus-6 has no apparent influence on course of HCV hepatitis, but may complicate HBV hepatitis and alcoholic liver disease. A pilot study. AB - Human herpesvirus-6 (HHV-6) is a widespread virus with occasional reactivation and a potential hepatotropism. The present study was undertaken to investigate the frequency of HHV-6 reactivation in viral (HCV, HBV) and alcoholic liver diseases and its implication for the course of the primary disease. Serological and immunohistochemical tests were done to document viral activity, hepatocellular apoptosis or proliferation, and autoantibody formation. While the course of HCV remains apparently uninfluenced by HHV-6, HBV hepatitis and alcoholic liver disease show a higher incidence of autoantibody formation if HHV 6 is present. The data of this pilot study warrant more extensive investigations of the clinical pathology of HHV-6 in liver diseases. PMID- 12655787 TI - An in utero allotransplantation model of metastatic breast cancer in the cat. AB - BACKGROUND: Breast cancer represents one of the most significant health risks in the human female population due both to its frequency and metastatic potential. Current therapies, while improving the overall survival rate of women with mammary neoplasias, have not eliminated this disease as an important cause of morbidity and mortality. MATERIALS AND METHODS: We have investigated transplantation of a feline mammary adenocarcinoma cell line into fetal cats as a model of human metastatic breast cancer. Fetal cats injected in utero with an allogeneic mammary adenocarcinoma cell line were born with palpable masses at the site of injection and developed widespread metastases over the following 6-10 weeks. Tumor foci were seen most commonly in the lung and in the local tissues adjacent to the primary injection site. This distribution of metastases mimics that seen in human breast cancer. Thus, in utero transplantation in cats is a reproducible experimental model of metastatic breast cancer in women. PMID- 12655788 TI - Evaluation of the morphology of submucosal tumor invasion and its volume in early gastric cancer. AB - BACKGROUND: The aim of the present study was to evaluate the clinical importance of the morphology of submucosal tumor invasion and its volume in early gastric cancer. MATERIALS AND METHODS: The subjects were 92 patients with a single lesion of early gastric cancer, who underwent gastrectomy with dissection of lymph nodes, and in whom the lesion was histologically-diagnosed as early gastric cancer with submucosal invasion. The volume of lesions (Vsm, S: < 50 mm3, L: > = 50 mm3) was determined by reconstructing submucosal lesions by the surface rendering method using pathological tissue sections (mean, 3.6 sections/lesion). The relationships between the volume of lesions and lymph node metastasis and between conventional clinicopathological parameters and lymph node metastasis were evaluated. RESULTS: Lymph node metastasis was observed in 18 patients (20%). The depth of submucosal invasion was 1.7 +/- 1.4 mm (mean +/- S.D.; range, 0.2 7.5 mm). sm1 (depth of submucosal tumor invasion < 0.5 mm) was observed in 11 patients (12%), and sm2 (depth of submucosal tumor invasion > = 0.5 mm) in 81 patients (88%). The mean Vsm was 104.1 +/- 215.4 mm3 (0.4-1,730.5 mm3). Fifty seven patients (62.0%) were in group S, and 35 patients (38.0%) in group L. There was a significant difference between lymph node metastasis and Vsm (S vs. L) alone (p = 0.006). Logistic regression analysis also demonstrated that Vsm alone was correlated with lymph node metastasis (p = 0.005; odds ratio, 1.27; 95% confidence interval, 0.074-0.401). CONCLUSION: The volume of submucosal lesions in early gastric cancer is important for evaluating lymph node metastasis. PMID- 12655789 TI - Altered tumor angiogenesis and metastasis of B16 melanoma in transgenic mice overexpressing tissue inhibitor of metalloproteinases-1. AB - Tissue inhibitor of metalloproteinases-1 (TIMP-1) has emerged as a multifunctional protein that plays contrasting roles during angiogenesis and cancer spread. We have investigated the growth, vascularization and metastasis of B16 melanoma cells in a transgenic mouse model with elevated TIMP-1 levels in the systemic circulation. Transgenic C57BL/6j-CBA mice overexpressing human TIMP-1 in the liver under the control of the mouse albumin promoter/enhancer were employed. An early subcutaneous growth advantage and an increased tumor angiogenic response were observed in transgenic animals with respect to wild-type hybrid mice. On the contrary, there was a dramatic decrease in the lung colonizing ability of B16 melanoma cells in TIMP-1 transgenic mice. No significant effect on metastasis formation was observed in another transgenic mouse model with increased TIMP-1 expression in lungs but low plasma levels, where the transgene was placed under the control of the murine mammary tumor virus promoter. These results support the notion that TIMP-1 displays paradoxical effects on tumor progression and suggest that circulating TIMP-1 is efficient in suppressing lung colonization of melanoma cells. PMID- 12655790 TI - Brucella melitensis infection stimulates an immune response leading to Kikuchi Fujimoto disease. AB - A 43-year-old Greek cattler with a history of brucellosis three months previously for which he was treated with sulfonamides for three weeks, was admitted to the hospital complaining of fever, arthralgias, night sweats, painful cervical and axillary lymph nodes as well as a weight loss of 8 kg in the previous four months. Since microbiological and serological studies did not give a specific diagnosis, an open cervical lymph node biopsy was performed. The histological examination revealed Kikuchi-Fujimoto disease. The etiology of the disease is unknown but viral, bacterial, protozoal and neoplastic as well as physicochemical agents may stimulate a particular immune response leading to Kikuchi-Fujimoto disease. Hereby, we present a case in which Kikuchi-Fujimoto disease followed brucella melitensis infection. This association permits us to hypothesize that the initial brucella melitensis infection three months previously triggered an immune response leading to Kikuchi-Fujimoto disease. The association of the disease with brucellosis is very important since these two entities share some similar characteristics, with brucellosis being relatively common in Europe. To our best knowledge, in the English language bibliography, this is the first reported case worldwide, though another similar case was described in the Spanish literature ten years ago. PMID- 12655791 TI - Review. DNA-mismatch repair and hereditary nonpolyposis colorectal cancer syndrome. PMID- 12655792 TI - Effects of high sucrose diet on body and liver weight and hepatic enzyme content and activity in the rat. AB - The effect of sucrose on the induction of hepatic and peripheral insulin resistance is well-documented. Studies show that, although oral administration of glucose does not significantly decrease total hepatic microsomal cytochrome P450 content, it causes an increase in cytosolic protein and in microsomal phospholipid and fatty acid content. In this study we examined the effects of a chronic high sucrose diet (HSD) on liver enzyme activity. Male Fisher 344 weanling rats were randomly assigned to a control diet (0% sucrose by calories, n = 10) or a diet in which starch was replaced by sucrose (65% sucrose, by calories, n = 10) for 90 days. The effects of HSD on weight gain, liver weight, hepatic microsomal cytochrome P450 (CYP450) content and glutathione-S-transferase (GST) activity were measured and compared with those fed standard lab chow. A small but statistically significant decrease in body weight (g) was seen in the sucrose-fed rats after day 50. Liver GST activity (nmol/mg protein/min) at the end of 90 days was decreased in animals maintained on HSD compared to those on the control diet, (181.7 +/- 8.0, 234.7 +/- 5.5), respectively. The liver weight and total CYP450 content in the two diet groups were not significantly different. The ratios of liver weight to body weight at the end of 90 days suggested that the livers of the HSD-fed animals were larger per gram of body weight. In addition, rats on the HSD had significantly smaller amounts of liver CYP450 1A1 and 3A2 than the rats on the control diet. These results suggest that a HSD may alter the hepatic enzyme activity which may affect the metabolism of substrates for these enzyme systems. PMID- 12655793 TI - A simple method to demonstrate normal and metaplastic Paneth cells in tissue sections. AB - Using hematoxylin and eosin (H&E)-stained sections, Paneth cells are identified by the presence of eosinophilic granules. Several other methods have been proposed to stain Paneth cells. Recently, we noticed that Paneth cells were autofluorescent in H&E sections. To assess the number of Paneth cells by this method, 30 surgical specimens from the small bowel and 10 from the colon with IBD were investigated. Consecutive sections were stained with H&E and immunostained with lysozyme (Lz). Paneth cell counting was done in 20 well-oriented, vertically cut crypts per-case using alternatively transmitted light (TL) and incident-light fluorescence (ILF). When H&E and Lz sections were observed with TL, the number of Paneth cells was easily assessed in some crypts, but in crypts having numerous Paneth cells, their exact number was ascertainable. On the other hand, using IFL, the number of H&E-stained Paneth cells-crypt was easily assessed, even in crypts with Paneth cell clustering. Using H&E-ILF, a mean of 3.9 (range 3-8) Paneth cells-pe-crypt was found in the normal small bowel and a mean of 3.3 (range 1-14) in the colon with IBD. ILF provided the adequate visual conditions--i.e. a dark, non-autofluorescent background--for assessing the actual number of normal and metaplastic Paneth cells, even when arranged in tight cell groups. The present method will improve the enumeration of the actual number of normal and metaplastic Paneth cells both in experimental research and in clinical trials. PMID- 12655794 TI - Clinical and biological effects of interleukin-2 with or without a concomitant administration of granulocyte-macrophage colony-stimulating factor in metastatic cancer patients. AB - GM-CSF has been shown to modulate the anticancer activity of IL-2 with, however, controversial results depending on the great variety of biological effects induced by GM-CSF itself. The activation of dendritic cells and the generation of suppressive cells would constitute the main favourable and unfavourable biological effects of GM-CSF, respectively. The present study was performed in an attempt to evaluate the clinical and biological effects of a concomitant GM-CSF administration of the immunotherapy of metastatic renal cell carcinoma with IL-2. The study included 25 patients, who were randomized to be treated with IL-2 alone or IL-2 plus GM-CSF. IL-2 was injected subcutaneously at 6 MIU/day for 6 days/week for 4 consecutive weeks, coressponding to one complete cycle. GM-CSF was injected subcutaneously at 0.3 micrograms/kg b.w. for 3 consecutive days for the first 3 days of each week of IL-2 administration. Two immunotherapeutic cycles at 21-day intervals were planned. No significant difference was observed in the percent of non-progressive disease between the two groups of patients. The increase in leukocyte mean number was significantly higher in patients concomitantly treated with GM-CSF, whereas no difference was observed in that of lymphocytes. This preliminary study suggests that the concomitant administration of GM-CSF does not enhance the therapeutic efficacy of IL-2 immunotherapy of metastatic renal cell cancer. PMID- 12655795 TI - Modification of radiation response in mice by ginsenosides, active components of Panax ginseng. AB - We performed this study to determine the effect of extract of whole ginseng and ginsenosides (total saponin, panaxadiol and panaxatriol) on jejunal crypt survival, endogenous spleen colony formation, and apoptosis in jejunal crypt cells of mice irradiated with high- and low-doses of gamma-radiation. The radioprotective effect of ginseng was compared with the effect of diethyldithiocarbamate (DDC). The jejunal crypts were protected by pretreatment with extract of whole ginseng (i.p.: 50 mg/kg of body weight at 12 and 36 hours before irradiation, p < 0.005). Extract of whole ginseng (p < 0.005), total saponin (p < 0.01) or panaxadiol (p < 0.05) administration before irradiation (i.p.: 50 mg/kg of body weight at 12 and 36 hours before irradiation) resulted in an increase in the formation of the endogenous spleen colony. The frequency of radiation-induced apoptosis in the intestinal crypt cells was also reduced by pretreatment with extract of whole ginseng (p < 0.05), total saponin (p < 0.005) or panaxadiol (p < 0.05) (i.p. at 12 and 36 hours before irradiation). The radioprotective effect on the jejunal crypts and apoptosis in the DDC-treated group appeared similar to that in the ginseng-treated groups. Treatment with DDC showed no significant modifying effects on the formation of the endogenous spleen colony. In the experiment on the effect of ginsenosides, the result indicated that panaxadiol might have a major radioprotective effect. Although the mechanisms of this inhibitory effect remain to be elucidated, these results indicated that ginseng might be a useful radioprotector, especially since it is a relatively nontoxic natural product. Further studies are needed to better characterize the protective nature of ginseng extract and each ginsenoside. PMID- 12655797 TI - An evaluation of the multiple dose protocol of cetrorelix in single ovary women. AB - BACKGROUND: The ovarian response and the pregnancy outcome of women with one ovary have been investigated under various IVF stimulation protocols. Cetrorelix is a novel agent offering important advantages in ovarian stimulation. In this study, the multiple protocol of cetrorelix is evaluated in single ovary women, for the first time. MATERIALS AND METHODS: Retrospective comparison of the ovarian response and ICSI/ET outcome between 12 cycles of single ovary women and 39 cycles of women with two ovaries. RESULTS: Between the two groups, there were no statistically significant differences in any of the analysed parameters. In the group of single ovary, 2 pregnancies were achieved and 4 children were born. In the group of two ovaries, 10 pregnancies were achieved and nine children were born. CONCLUSION: Single ovary women have the same response to ovarian stimulation with cetrorelix/hMG/rFSH as women having both ovaries, but they present a lower, although not statistically significant, pregnancy rate. PMID- 12655796 TI - Eicosanoid precursor polyenoic fatty acids modulate synaptic levels of dopamine in ex-vivo slices of rat brain striatum. AB - BACKGROUND: Considerable evidence indicates that polyunsaturated fatty acids are important in normal brain structure and function. MATERIALS AND METHODS: Rat brain striatal slices incubated with tritiated dopamine were electrically stimulated twice. During the first only buffer was perfused. During the second period buffer, fatty acid plus indomethacin, or fatty acid plus nordihydroguaiaretic acid were perfused. The ratio of the two stimulations indicated changes in released tritium. RESULTS: The only fatty acids to induce significant changes in tritium were the eicosanoid-precursors, dihomo-gamma linolenic, arachidonic and eicosapentaenoic acids. DISCUSSION: There were no differences between the effects of the fatty acid alone or fatty acid in the presence of indomethacin, indicating little involvement of the cyclooxygenase pathway. Fatty acid in the presence of nordihydroguaiaretic acid reversed the low synaptic tritium concentrations, indicating that the lipoxygenase pathway may be active in dopaminergic metabolism in striatum. PMID- 12655798 TI - Ki-67, cyclin D1, p53 and bcl-2 expression in advanced head and neck cancer. AB - In order to analyse the radiosensitivity of tumours and to evaluate the possibility of improving the treatment results with regard to genetic alterations, we examined 33 patients with advanced head and neck tumours after 66 70 Gy irradiation. Between 1998 and 2001, 33 patients with advanced head and neck squamous cell carcinoma (HN-SCC) were observed. They received 66-70 Gy to the primary tumour site and pathological lymph nodes. One month later, physical examination and CT were performed to verify the effect of radiotherapy, and the patients were followed until their death. The histological grading and Ki-67, cyclin D1, p53 and bcl-2 status were examined from the aspect of their potential prognostic value in all patients. The average survival was 13 months; 25% of the patients survived for at least 20 months. Seventy-two percent of the patients demonstrated Ki-67 positivity, 69% p53 positivity and 40% cyclin D1 positivity; there were only 12% bcl-2-positive cases. A significant correlation was not found between the tumour response or the duration of survival and the Ki-67, p53 or cyclin D1 positivity. Only bcl-2-positive cases exhibited significantly better outcome. These parameters indicate the proliferating (Ki-67 and cyclin D1) and apoptotic (p53 and bcl-2) activities of advanced HN-SCC cells. Our results proved that they proliferate rapidly and have impaired repair or apoptotic functions. The heterogeneity of our results did not allow us to conclude that the above parameters are of clinically reliable prognostic value, but the obviously high kinetic activity of HN-SCC underlines the potential efficacy and need for accelerated irradiation in these cases. PMID- 12655799 TI - Possible mechanism of local tumor progression in experimentally-induced colorectal carcinomas in rats. AB - Sections from 130 DMH-induced carcinomas in rats were reviewed. In 78% (74 out of 95) of the gland-forming adenocarcinomas, neoplastic glands displayed at the growing edge either a thinner layer or a partial loss of tumor cells ("pore" formation). The thinner epithelium appears to be a stage preceding the focal "destruction" of the tumor cells ("pore" formation). Through those glandular "pores" mucins, inflammatory cells or necrotic elements were being released free into the surrounding host tissues. It is theorized that those non-neoplastic "pseudopodial" forerunners (rich in proteolytic enzymes) encourage the degradation of the surrounding host tissues. The malignant cells surrounding glandular "pores" eventually grow around the secreted products to restore the continuity of the gland. The remodelling of the glands will guarantee a stepwise but everlasting tumor progression deeper into the host. PMID- 12655801 TI - Perspectives. Budget: vaccine program gets booster shot; NIH star fades. PMID- 12655800 TI - Effects of Cupressus sempervirens cone extract on lipid parameters in Wistar rats. AB - BACKGROUND: Lipid-lowering drugs have been developed to treat hyperlipidemia, but they are expensive, while serious side-effects may occasionally occur. In Albania, a hydroalcoholic extract of the cones of Cupressus sempervirens is used to treat a variety of disorders, including hyperlipidemia. The purpose of our study was to assess the effects of Cupressus sempervirens cone extract (CSE) on the lipid profile of Wistar rats. MATERIALS AND METHODS: The animals were divided into two groups. Group I (30 animals) served as a control group. Serum lipid parameters, muscle and liver enzymes, red and white blood count, platelets, and serum concentrations of uric acid and creatinine were determined at baseline and at weeks 6, 12, 18 and 24 of the study. RESULTS: The oral administration of the extract resulted in a substantial decrease of serum total cholesterol, which was significant even after 6 weeks of treatment. Moreover, these animals exhibited lower total cholesterol levels compared to the controls after the initiation of treatment (p < 0.001) during the study period. The administration of the extract led to a substantial reduction in serum triglycerides (p < 0.05) in Group I, comparing 0 week to 6-24 weeks. However no significant differences in triglyceride levels were observed between CSE animals (Group I) and controls (Group II) during the entire study period. No significant changes in HDL cholesterol in the other parameters occurred in either group. CONCLUSION: The administration of CSE has an important lipid-lowering effect in Wistar rats. PMID- 12655802 TI - Biosorption of cadmium and chromium in duckweed Wolffia globosa. AB - The biosorption of cadmium (Cd) and chromium (Cr) by using dried Wolffia globosa biomass were investigated using batch technique. The effects of concentration and pH solution on the adsorption isotherm were measured by determining the adsorption isotherm at initial metal concentrations from 10 to 400 mg/L and pH 4 to 7 for Cd, and pH 1.5 to 6 for Cr. The adsorption equilibria were found to follow Langmuir models. The maximum adsorption capacity (Xm) at pH 7 in W. globosa-Cd system was estimated to be 80.7 mg/g, while the maximum removal achieved at pH 4, pH 5, and pH 6 were 35.1, 48.8, and 65.4 mg/g, respectively. The Xm at pH 1.5 in W. globosa--Cr system was estimated to be 73.5 mg/g, while the maximum removal achieved at pH 3, pH 5, and pH 6 were 47.4, 33.1, and 12.9 mg/g, respectively. The effects of contact times on Cd and Cr sorption indicated that they were absorbed rapidly and more efficiently at lower concentrations. PMID- 12655803 TI - Phytoaccumulation and phytotoxicity of cadmium and chromium in duckweed Wolffia globosa. AB - The phytoaccumulation and phytotoxicity of heavy metals, cadmium (Cd), and chromium (Cr) on a common duckweed, Wolffia globosa, were studied. W. globosa were cultured in 3% Hoagland's nutrient medium, which was supplemented with 1, 2, 4, and 8 mg/L of Cd and Cr and were separately harvested after 3, 6, 9, and 12 days. The accumulation of Cd and Cr in W. globosa showed significant increases when the exposure time and metal concentration were increased. The effects of Cd and Cr on the biomass productivity and total chlorophyll content in W. globosa indicated that there were significant decreases in the biomass productivity and total chlorophyll content when the exposure time and metal concentration were increased. PMID- 12655804 TI - Isolation, characterization, and identification of bacteria associated with the zinc hyperaccumulator Thlaspi caerulescens subsp. calaminaria. AB - We investigated bacterial populations associated with the Zn hyperaccumulator Thlaspi caerulescens subsp. calaminaria grown in a soil collected from an abandoned Zn-Pb mine and smelter in Plombieres, Belgium. The bacterial population of the nonrhizospheric soil consisted of typical soil bacteria, some exhibiting multiple heavy-metal resistance characteristics that often are associated with polluted substrates: 7.8% and 4% of the population survived in the presence of elevated levels of Zn (1 mM) and Cd (0.8 mM), respectively. For the bacterial population isolated from the rhizosphere, the comparable survival rates were 88 and 78%. This observation indicates a selective enrichment of the metal-resistant strains due to an increased availability of the metals in soils near the roots compared with nonrhizospheric soil. The endophytic inhabitants of the roots and shoots were isolated, identified, and characterized. Although similar endophytic species were isolated from both compartments, those from the rhizoplane and roots showed lower resistance to Zn and Cd than the endophytic bacteria isolated from the shoots. In addition, root endophytic bacteria had additional requirements. Contrary to the rootresiding inhabitants, the shoot represented a niche rich in metal-resistant bacteria and even seemed to contain species that were exclusively abundant there. These differences in the characteristics of the bacterial microflora associated with T. caerulescens might possibly reflect altered metal speciation in the different soils and plant compartments studied. PMID- 12655805 TI - Bioengineering of a phytoremediation plant by means of somatic hybridization. AB - Phytoremediation is a technology that exploits a plant's ability to remove contaminants from the environment or render toxic compounds harmless. An efficient metal phytoremediating plant must combine high biomass production and established cultivation methods with high tolerance to a specific contaminant and ability for root uptake, translocation, and compartmentalization of contaminants in the above-ground biomass. Symmetric and asymmetric somatic hybridizations were used to introduce toxic metal-resistant traits from Thlaspi caerulescens into Brassica juncea. B. juncea hypocotyl protoplasts were fused with T. caerulescens mesophyll protoplasts. The hypocotyl protoplasts of B. juncea were stained with CFDA before fusion and thus fluoresced green under UV, whereas the mesophyll protoplasts of T. caerulescens had red autofluorescense. Heteroplasmic fusion products were identified and selected by flow cytometry and cell sorting. All putative hybrids grown in the greenhouse had morphological characteristics of B. juncea. A Thlaspi-specific repetitive sequence was hybridized to total DNA of plants, including the parental species. All plants from both symmetric and asymmetric fusions showed Thlaspi-specific hybridization patterns while B. juncea did not exhibit any hybridization signal. Hybrid plants, produced by asymmetric somatic hybridization between the two species, demonstrated high metal accumulation potential, tolerance to toxic metals, and good biomass production. PMID- 12655806 TI - Removal of N, P, BOD5, and coliform in pilot-scale constructed wetland systems. AB - Pilot-scale surface-flow (SF), subsurface-flow (SSF), and floating aquatic plant (FAP) constructed wetland system designs were installed and evaluated to determine the effectiveness of constructed wetlands to treat tertiary effluent wastewater in a Midwestern U.S. climate (central Illinois). Average ammonia nitrogen (N) concentrations decreased approximately 50% in the SSF system design, suggesting that this design had the highest nitrification rate. Nitrate-N concentrations decreased by over 60% in the FAP system design, possibly due to dissimilatory reduction or plant uptake. Total phosphorus (P) concentration reductions of 25 to 40% were observed in all three system designs. Five-day biochemical oxygen demand (BOD5) and dissolved oxygen (DO) results suggested that biodegradation was highest in the SSF system design and lowest in the FAP system design. Greater than 90% concentration reductions of total coliform and E. coli recovered were also observed following treatment in all three system designs. The FAP system design appeared to yield the highest concentration reduction efficiency for E. coli, possibly due to increased sunlight and related bacteriocidal ultraviolet light exposure. Ongoing experiments will test regularly for a variety of vegetative, water quality, and biological conditions for longer time periods in order to gain a better understanding of the pilot constructed wetland system design kinetics. PMID- 12655807 TI - Treatment of munitions in soils using phytoslurries. AB - Phytoremediation is an established technology for the treatment of explosives in water and soil. This study investigated the possibility of using slurried plants (or phytoslurries) to treat explosives (TNT and RDX). The degradation of TNT in solution using intact and slurried parrotfeather (Myriophyllum aquaticum), spinach (Spinicia oleracea), and mustard greens (Brassica juncea) was evaluated. Phytoslurries of parrotfeather and spinach removed the TNT faster than the intact plant. Conversely, the removal rate constants for slurried and intact mustard greens were about the same. A study using pressurized heating to destroy enzymatic activity in the phytoslurries was also conducted to compare removal from released plant chemicals to adsorptive removal. Aqueous phase removal of TNT by autoclaved spinach phytoslurry was compared with nonautoclaved spinach phytoslurry. The autoclaved phytoslurry did remove TNT, but not as completely as nonautoclaved slurry. This suggests that some removal is due to adsorption, but not all. Phytoslurries of mustard greens and parrotfeather had higher RDX removal rates compared with intact plant removal, but the rates for parrotfeather in either case were relatively low. Phytoslurries of spinach had relatively modest increases in RDX removal rates compared with intact plant. Studies were then conducted with phytoslurry/soil mixtures at two scales: 60 ml and 1.5 l. In both cases, phytoslurries of mustard greens and spinach removed TNT and RDX at higher levels than control slurries. PMID- 12655808 TI - Remediation of benzo[a]pyrene and chrysene-contaminated soil with industrial hemp (Cannabis sativa). AB - The phytoremediation, with industrial hemp (Cannabis sativa), of a Hawaiian silty clay soil contaminated with two polycyclic aromatic hydrocarbons (PAHs), chrysene and benzo[a]pyrene, was studied. Hemp showed a very high tolerance to the contaminants. The growth rates of hemp, compared with control, in soils fortified with chrysene and benzo[a]pyrene at concentrations of each varying from 25 to 200 micrograms/g were consistently above 100%. The plants grew from seed for 45 days in soil fortified with PAHs at concentrations of 25, 50, and 75 micrograms/g. Controls were pots with contaminated soil but no plant. PAHs levels were significantly reduced in all pots (control and seeded pots), expect for one set at a high concentration of chrysene, which may be due to uneven spiking. A time course study over 28 days was done to monitor changes of microbial count and levels of chrysene. Little changes were observed for the total microbial count in the soil, and the concentration of chrysene in the soil decreased slightly in the pots containing plants. However, the chrysene levels in those pots were consistently lower than those in the pots without plants. PMID- 12655810 TI - A comparison of a threat of smallpox in the United States and Pakistan from a student perspective. PMID- 12655811 TI - Infectious disease (Ebola virus). PMID- 12655812 TI - A cancer clip. PMID- 12655814 TI - [Home care services, a reaffirmed role in the health system]. PMID- 12655816 TI - [The war against AIDS continues!]. PMID- 12655817 TI - [For an active prevention of cardiovascular diseases]. PMID- 12655818 TI - [Chronic inflammatory bowel diseases come out of the shadows]. PMID- 12655819 TI - [Use of a blood glucose monitor in a specialized hospital unit]. PMID- 12655821 TI - [Rare diseases, a public health problem]. PMID- 12655820 TI - [Skin preparation in surgical patients, study of practices in a hospital center]. PMID- 12655822 TI - [Ethics and rare diseases]. PMID- 12655823 TI - [Nursing care and support for patients with rare diseases]. PMID- 12655824 TI - [Jeremy, afflicted with acute idiopathic hypersomnia]. PMID- 12655825 TI - [Orphanet, an information site on rare diseases]. PMID- 12655826 TI - [The French Myopathy Association's campaign against emerging rare diseases]. PMID- 12655827 TI - [Care in the treatment of orphan diseases at home]. PMID- 12655828 TI - [9/9 Moral harassment]. PMID- 12655829 TI - HIV-positive gay and bisexual men: predictors of unsafe sex. AB - This paper describes the sexual risk practices of 322 HIV-positive gay and bisexual men who reported having unprotected anal, oral or vaginal sex with an HIV-negative or unknown status partner(s) during the previous four months. Descriptive data on type of sex, partner types and number of partners are presented. Determinants of unprotected anal and oral intercourse were examined using a social cognitive framework. More unprotected anal sex was associated with having a steady versus a casual partner, not being on triple drug therapy, increased use of drugs and alcohol, and lower outcome expectancies for condom use and negotiation of safer sex practices. Unprotected oral sex was associated with partner type; men with casual partners had less unprotected oral sex as compared to men with steady partners. Findings are discussed in relation to the development of sexual risk reduction interventions for HIV-positive individuals. PMID- 12655830 TI - Childhood sexual abuse and sexual transmission risk behaviour among HIV-positive men who have sex with men. AB - Previous studies have indicated an association between childhood sexual abuse (CSA) and adult sexual risk behaviour among women and among men who have sex with men (MSM). However, no studies to date have tested the hypothesis that a history of CSA predicts sexual behaviour carrying risk of transmission of HIV to others, i.e. in a known HIV-positive cohort. The present study tested this hypothesis among a sample of 456 HIV-positive MSM recruited from community venues in New York and San Francisco. CSA history was found to be significantly associated with past (in the last 90 days) unprotected anal sex acts, both insertive (33% versus 20%, p < 0.05) and receptive (43% versus 27%, p < 0.02), with partners of HIV negative or unknown serostatus. Further, several potential mediators of this effect were tested, and three found to be predicted by CSA history. Each of these potential mediators was associated with sexual risk behaviour, but differentially: anxiety and hostility were significantly associated with insertive acts, while anxiety, hostility and suicidality were associated with receptive acts. Mediation analyses supported the hypothesis that these factors significantly (albeit partially) accounted for the association of CSA with receptive anal intercourse. Nonsignificant mediation effects were found for insertive sex, suggesting the operation of unmeasured mediating variables. These results highlight the importance of mental health services for individuals who have been sexually abused, both for personal and for public health benefit, and also indicate a need for further research into mediators of CSA effects on transmission-related behaviour. PMID- 12655832 TI - Leaving sex work: barriers, facilitating factors and consequences for female sex workers in northern Thailand. AB - Factors facilitating or inhibiting women's ability to leave sex work are still poorly characterized, and little is known about women's lives after they leave the profession. This paper presents findings from a qualitative study about factors affecting women's ability to leave sex work and influencing their lives after leaving. We interviewed 42 current and former female sex workers (FSWs) drawn from a cohort study of 500 FSWs in northern Thailand. All but one of the participants had quit sex work at least once. The majority experienced one or more quit-re-entry-quit cycles. Women's ability and decisions to leave sex work were determined primarily by four factors: economic situation, relationship with a steady partner, attitudes towards sex work and HIV/AIDS experience. Economic concerns, ranging from survival needs to materialistic desires, had the strongest influence. Most women perceived their risk for HIV infection to be lower after leaving sex work, but three of the 17 HIV-infected women acquired infection after having left, presumably from their steady partners. Prevention efforts should guide women as they transition out of commercial sex work. Interventions aimed at assisting women wanting to leave sex work need to address the role of economic factors. PMID- 12655831 TI - Availability of combination therapy for HIV: effects on sexual risk taking in a sample of high-risk gay and bisexual men. AB - The objective of this study was to assess the impact of successes in the medical treatment of HIV disease on the perceptions of high-risk gay and bisexual men attending an anonymous testing site towards becoming infected with HIV and on their decisions to have unsafe sex. A cross-sectional survey with a face-to-face interview and self-recorded information was used. A convenience sample of high risk men seeking anonymous HIV antibody counselling was recruited for an intervention study. To be eligible, men had to have a history of at least one previous negative antibody test and unprotected anal intercourse within the last year. Upon enrollment, subjects were asked about the impact of new treatment developments on their decisions to engage in high-risk sex both in the recent past and in the future. In general, neither the recent successes in treating HIV nor the availability of combination therapy had any impact on the men's decisions about safe sex, although a minority of men reported less concern about becoming HIV-positive. Also, some men reported already having taken more risk. In conclusion, a small number of men in this high-risk population have increased risk behaviour as a result of recent treatment advances. These results merit continued monitoring. PMID- 12655833 TI - HIV prevalence and sexual behaviour of male clients of brothels' prostitutes in Dakar, Senegal. AB - This study reports the results of research designed to determine the prevalence of HIV infection in a group of male clients of brothel prostitutes, and to describe characteristics associated with HIV infection in Dakar, Senegal. Clients come from the lower portion of the social scale rather than from the middle class or from the wealthier groups of the population. A significant number of these men did not use condoms with their regular partners, and another group reported sexual contact with occasional partners with whom condom use was not usual. Additionally, for a fraction, condoms were not systematically used either with prostitutes or with partners. HIV prevalence in clients appeared to be much higher than the prevalence in the general population and HIV-infected clients were older than HIV-negative clients. The significant association between HIV infection and age can be explained by the fact that older clients probably have been exposed to prostitution longer. HIV-positive clients had more contact with prostitutes during the previous seven days, and in addition they also had more occasional sexual encounters than did the HIV-negative clients. This demonstrates that a multiplicity of sexual partners increases the risk of HIV infection beyond the contact with prostitutes for this group of men. PMID- 12655834 TI - HIV/AIDS knowledge, attitudes, and practices among Burmese migrant factory workers in Tak Province, Thailand. AB - Little information is known regarding the level of knowledge among Burmese migrant workers along the Thai/Burma border concerning risk factors, prevention or transmission of HIV. The Burma Medical Association (BMA) and National Health and Education Committee (NHEC) collected data on knowledge, attitudes and practices among 725 factory workers in Tak Province, Thailand during July 2000. We present results from a secondary analysis of these data. Responses were grouped into prevention, transmission and risk categories, and percentages answered correctly were recorded. Men consistently scored higher than women, with significant gender differences in the prevention and transmission questions. Forty-one per cent of the women understood that contraceptive pills do not prevent infection and 15% of females reported ever seeing a condom. Twelve per cent of men and 1.4% of women reported ever using a condom (OR = 8.57, p < 0.0001). Previously, virtually no access has been extended to persons trying to document health status among Burmese migrant workers in factories. The survey reveals a significant lack of knowledge about HIV among factory workers and indicates that a sub-population of Burmese people appears to lack the most basic information about the epidemic. PMID- 12655835 TI - Study of the effect of information, motivation and behavioural skills (IMB) intervention in changing AIDS risk behaviour in female university students. AB - This study has perhaps for the first time in the Indian context made an attempt to use a psychosocial model for a HIV/AIDS intervention programme. In line with the same, the main objective of the research was to study the effect of the information, motivation and behavioural skills (IMB) model-based intervention on a group of female college/university students. Purposive sampling was used to select 200 participants; pre-testing of these participants revealed that they did not have adequate information on issues related to HIV transmission and prevention. Further, neither were they motivated nor possessed appropriate behavioural skills to engage in HIV preventive behaviours. Following this, 100 participants were assigned to the experimental group and the remaining 100 to the control group. Post-testing showed a significant increase in the level of information, motivation and behavioural skills of the experimental group participants who underwent the three-session intervention programme as compared to the control group participants. PMID- 12655836 TI - HIV prevention in Latin America: reaching youth in Colombia. AB - The aim of this paper is to describe and evaluate a school-based peer education programme on HIV primary prevention implemented in urban marginal districts of three cities of Colombia from 1997 to 1999. Its main objective was to promote risk awareness and safe sexual behaviours among urban youth populations. Methodology included the collection of baseline information through qualitative methods (focus groups and in-depth interviews), a knowledge, attitudes and practices (KAP) survey, a health education intervention, and post-intervention data collection. Direct beneficiaries were adolescents 10 to 19 years of age, and secondary school teachers of 6th to 9th grades. Main strategies used were peer education and classroom sessions conducted by trained teachers. Short-term results suggest that the programme had a positive effect on knowledge and attitudes related to HIV/AIDS (as the mean knowledge summary indicator among adolescents and secondary school teachers increased 24% and 21%, respectively). The main outcome has been the development of a sex education programme, emphasizing the role of schools in the promotion of sexual and reproductive health. Mass education by a combination of interventions and events at school level, backed up by effective interpersonal communication such as peer education, classroom teaching and community actions are effective primary prevention strategies for HIV sexual transmission and should be more extensively considered. PMID- 12655837 TI - Characteristics of HIV antiretroviral treatments, access and adherence in an ethnically diverse sample of men who have sex with men. AB - Data regarding HIV antiretroviral treatment regimens, access to treatment and medical care, and adherence to medications were collected as part of the Seropositive Urban Men's Study, a formative study of HIV-positive men who have sex with men. Participants (N = 456) were recruited from AIDS service organizations, mainstream gay venues and public/commercial sex environments. The sample was 94% gay or bisexually-identified; 29% were African American, 24% Latino and 30% white. The majority (71%) indicated being on antiretroviral treatment, and most were taking a protease inhibitor/nucleoside reverse transcriptase inhibitor combination. African American men in New York City were less likely to be on treatment. Among those on treatment (n = 322), 51% reported at least one day in which they had missed a dose of their medication and the mean number of days in which a dose was missed (in the past 30 days) was 1.72. Multivariate analyses indicated that avoidant coping, frequency of drinking alcohol and difficulty in communicating with sex partners about HIV were related to days of missed doses, suggesting the need or desire to escape from the reality of life with HIV as a potential explanation for poor adherence. PMID- 12655838 TI - Home-based study of anti-HIV drug regimen adherence among HIV-infected women: feasibility and preliminary results. AB - Collection of antiretroviral medication adherence data in the homes of HIV infected people may have methodological advantages that can improve data quality. However, the feasibility of this approach has not been established. In addition, data on adherence, and its predictors, among HIV-infected women have been limited. Sixty-three HIV-positive women who were prescribed at least one antiretroviral drug in the last month were interviewed in their homes. A standard instrument was used to collect data on all antiretroviral medications prescribed and taken in the three days prior to the interview. Data were also collected on factors thought potentially to affect the ability to be adherent. The results of this study suggest that it is feasible to conduct home-based adherence research. Sixty-seven per cent reported taking all prescribed antiretroviral medication doses. One-third took a sub-optimal dose putting themselves at increased risk of treatment failure and the selection of resistant HIV strains. Unintentional reasons for missing doses were most commonly reported. An ability to describe the intended effect of antiretroviral therapy on HIV viral load was the best predictor of adherence. This finding is consistent with other research suggesting that adherence is associated with an understanding and belief in the effectiveness of antiretroviral therapy. PMID- 12655839 TI - The problem of antiretroviral adherence: a self-regulatory model for intervention. AB - Controlled trials that determine which interventions can significantly improve adherence to combination antiretroviral regimens are urgently needed. Rates of adherence to the regimens are often sub-optimal, and while a variety of factors that are predictive or associated with antiretroviral adherence have been identified, few research findings are reported of interventions that effectively promote adherence and improve health outcomes. Further, the mechanisms by which interventions promote antiretroviral adherence are not well understood. Maximizing adherence and achieving the full potential of the antiretroviral therapies demand multidimensional initiatives that address complex behavioural and biomedical issues. A model drawn from self-regulation theory and empirical data to explain antiretroviral medication adherence is proposed. PMID- 12655841 TI - UNAIDS/WHO global AIDS statistics. PMID- 12655840 TI - A pilot study of the effects of a behavioural intervention on treatment adherence in HIV-infected patients. AB - A pilot study was carried out in a group of six HIV-infected non-adherent men testing the effects of a behavioural medication management intervention on adherence with antiretroviral drugs. The study was prospective, using a one-group repeated-measures design. Adherence was measured using two self-reports. The intervention was a behaviourally-based programme that lasted for three months and included individualized education about antiretroviral medication and their side effects; positive reinforcement and encouragement; individualized counselling weekly; follow-up calls; and lifestyle assessment and the identification of adherence barriers. Assessments were carried out at recruitment, immediately after the initiation of the intervention, one month, three months and six months later. Results suggested that the intervention enhanced adherence rates from a mean percentage of 80.27 at baseline to a mean of 97.5% at the end of follow-up (six months time point). Similar improvement was observed in the scores of the scale measuring adherence difficulties. Further, CD4+ counts also slightly improved. Interviews with four of the participants at the end of the study highlighted the problems experienced by patients in taking their medication and supported the usefulness of the intervention. Because of the complexity of the factors behind adherence, it is important that patients are supported with individualized medication management programmes. PMID- 12655842 TI - Sensitive magnetic sensors without cooling in biomedical engineering. AB - Magnetic field sensors are used in various fields of technology. In the past few years a large variety of magnetic field sensors has been established and the performance of these sensors has been improved enormously. In this review article all recent developments in the area of sensitive magnetic field sensory analysis (resolution better than 1 nT) are presented and examined regarding their parameters. This is mainly done under the aspect of application fields in biomedical engineering. A comparison of all commercial and available sensitive magnetic field sensors shows current and prospective ranges of application. PMID- 12655844 TI - [Development of a periodontium-approximated specimen holder for attrition studies of telescopic crowns]. AB - To enable attritional studies on telescopic crowns, a specimen holder has been constructed that is intended to simulate, in zero approximation, the biomechanical properties of abutments bearing telescopic crowns. The specimen holder consists essentially of a system with two metal "concertinas" with different elasticity constants reflecting the different biomechanical parameters for periodontium and bone. The biomechanical processes in the periodontium are too complex to be simulated accurately in vitro. Nevertheless, the specimen holder described here enables the forces acting during insertion and removal of a partial denture to be accommodated elastically in the angular and axial directions, to simulate the possible degrees of freedom of the abutments in situ. In this way, the unavoidable errors of specimen fixation are rendered tolerable, so that local preferential wear is decisively reduced. PMID- 12655843 TI - Strain measurement at the knee ligament insertion sites. AB - We describe the modification of an existing method of ligament strain measurement at the knee joint in detail. At ten fresh joint specimens we used that technique where strain gauges are attached to the ligamentous insertions and origins. We both improved the preparation of the attachment site and the application of the strain gauges. In a special apparatus the specimens were moved from 0 degree extension to 100 degrees flexion while simulating muscle strength and axial force. Testing was performed at the posterior cruciate ligament with both intact and transsected anterior cruciate ligament. In contrast to other existing techniques it does not affect the motion of the joint or the integrity and the function of the ligaments. Unlike the original description of that method we could register a loading behaviour of the posterior cruciate ligament that is similar to those reported in the literature. PMID- 12655845 TI - [Transmission electron microscopy image of wear particles of joint endoprostheses and ultrastructural cell changes]. AB - Wear particles from joint endoprostheses vary considerably in size, and may be detectable in tissue only by electron microscopy. Wear debris plays a central role in the non-infectious late loosening of prostheses, and it has been estimated that the submicron particles induce increased liberation of mediators of osteolysis by activated macrophages. From the types of prostheses currently in use, bone cement and polyethylene particles greatly predominate over metallic and ceramic particles. Since it had formerly not been possible to reliably identify wear particles in the transmission electron microscopy, and descriptions of them in the literature varied considerably, we analysed ultrathin sections obtained from periprosthetic tissue containing wear particles previously identified by laser microprobe mass analysis. Using this method, it proved possible to classify almost all the wear particles detected in the electron microscope, to determine their size range and to represent the cellular alterations caused by them. PMID- 12655846 TI - [Microdialysis as a procedure for evaluating intestinal hypoxia--an animal experiment approach]. AB - Usual ICU monitoring of patients with abdominal pathology provides no detailed information about hepatosplanchnic haemodynamics or intestinal metabolism. In our animal experiment, the effects of systemic hypoxia on microdialysis measurements of the peritoneum in comparison with the ischiocrural muscle as reference were investigated in 7 rats. The parameter of interest was the course of glucose metabolism reflecting sympathoadrenergic activity during the experiment. Measurements were obtained at timed intervals at baseline, under hypoxia, and during reoxygenation. After induction of systemic hypoxia, the peritoneal microdialysis showed significantly higher levels of glucose in comparison with the ischiocrural muscle. The results indicate hypermetabolic activity or a hypersympathetic response of the bowel in response to hypoxic stress. In the clinical setting, the bowel has an important role in the development of multiorgan failure. Microdialysis may therefore be an interesting tool for the early detection of hypoxic metabolism during and after abdominal procedures. PMID- 12655848 TI - Working together--but against the odds. PMID- 12655847 TI - Frequency component selection for an ECoG-based brain-computer interface. AB - The aim of the present study was to investigate the most significant frequency components in electrocorticogram (ECoG) recordings in order to operate a brain computer interface (BCI). For this purpose the time-frequency ERD/ERS map and the distinction sensitive learning vector quantization (DSLVQ) are applied to ECoG from three subjects, recorded during a self-paced finger movement. The results show that the ERD/ERS pattern found in ECoG generally matches the ERD/ERS pattern found in EEG recordings, but has an increased prevalence of frequency components in the beta range. PMID- 12655849 TI - Scepticism over AIDS claim. PMID- 12655850 TI - Counting on co-operation. PMID- 12655851 TI - Learning how to work together. PMID- 12655852 TI - Learning how to work together. PMID- 12655853 TI - Delay tactics. PMID- 12655855 TI - Partnership of equals. PMID- 12655854 TI - Shooting pains. PMID- 12655856 TI - Testing times. PMID- 12655857 TI - Mind your language. PMID- 12655858 TI - An evaluation of the Leading an Empowered Organisation programme. AB - AIM: To evaluate outcomes from the NHS leadership development programme Leading an Empowered Organisation (LEO). METHOD: A prospective case study based on a pre- and post-course quantitative analysis with interpretative methodological support. RESULTS: Role conflict, whether nurse or manager, was apparent in nurses' perceptions of their roles. Respondents claimed many positive leadership attributes but lacked assertiveness and the skills for handling conflict. The LEO programme had a statistically significant effect on workplace leadership performance and positive benefits related to communication competence, articulation of goals, networking, assertiveness, zones of responsibility and problem solving. CONCLUSION: LEO is having an effect on workplace performance. However it could be improved by considering the package holistically, including not only the course content, but the entry level, pre-course preparation and post course mentorship. PMID- 12655859 TI - Continuing professional development: a review. AB - In the UK health sector, continuing professional development for nurses is now clearly identified as necessary to the continued improvement of clinical services. The clinical governance movement that is driving this improvement has had a great effect on the NHS and it is now necessary to demonstrate service maintenance and improvement. Lifelong learning and the enhancement of skills through training or self-development are mentioned as ways of providing staff governance (DoH 1999, SEDH 1999). PMID- 12655860 TI - Measles: the disease and its prevention. AB - The MMR vaccine is currently the subject of controversy. Helen Bedford describes the measles disease and its complications, and advises nurses on what to tell parents about vaccination. PMID- 12655861 TI - Investing in people. PMID- 12655863 TI - Away with the fairies. PMID- 12655862 TI - Support systems. PMID- 12655864 TI - Primary care. Where next? PMID- 12655865 TI - Winner by smiles. Interview by Nick Edwards. PMID- 12655866 TI - Food procurement. Making a meal of it. AB - The NHS needs to explore alternative approaches to food procurement in the interests of good nutrition and waste reduction. Other European countries have interpreted EU regulations on public sector procurement in ways which allow them to buy local produce. Many trusts are unsure about their powers to implement changes in food procurement. PMID- 12655867 TI - Joint-working. Worlds apart. AB - A programme bringing together primary care trust non-executives and local counsellors from the same patch changed participants' perceptions of each other. PCT non-executives and counsellors were convinced of the benefits of joint working, but unsure how to go about it. Knowledge of each other's worlds was initially poor. Annual structured visits between PCTs and local authorities should be encouraged. PMID- 12655868 TI - Human resources. Grade expectations. PMID- 12655869 TI - Room to move. PMID- 12655870 TI - Research governance. Checking it out. PMID- 12655871 TI - Reducing health inequalities. Long division. PMID- 12655872 TI - Reducing health inequalities. Local care for local people. PMID- 12655874 TI - Reducing health inequalities. Public property. PMID- 12655873 TI - Reducing health inequalities. Teenage sex: co-ordination counts. PMID- 12655875 TI - Hunger pains. Diet: mapping local access to healthy food. PMID- 12655877 TI - No butts.... Smoking cessation: working with authority figures in ethnic minority communities. PMID- 12655878 TI - Telling our story. ACHE launches a national image campaign to promote the role of healthcare executives. PMID- 12655879 TI - Interview with ACHE's incoming chairman. Interview by Ellen G. Lanser. PMID- 12655880 TI - Surviving the medical malpractice crisis. PMID- 12655881 TI - Hospital/physician relations: collaboration or competition? PMID- 12655883 TI - Ensuring your ROMI. Does your service contract help you achieve a return on your maintenance investment? PMID- 12655884 TI - Evaluating claims of conscience. With appropriate guidelines, you can respect staff's beliefs while providing quality care for patients. PMID- 12655885 TI - Readers' picks. Young healthcare executives highlight the Web sites that best meet their needs. PMID- 12655886 TI - The hard case for soft skills and retention. Staffing woes may be related to poorly developed soft skills in your leaders. PMID- 12655887 TI - Long-distance mentoring. Make the most of mentoring when you can't meet in person. PMID- 12655888 TI - Recovering physician loyalty. Lessons from a national physician survey on crafting a true hospital-physician partnership. PMID- 12655889 TI - Community redevelopment. A health system helps to revitalize neighbourhoods. PMID- 12655890 TI - The rising cost of healthcare. Cost increases drive healthcare to the top of the domestic policy agenda. PMID- 12655891 TI - The view from the middle. Successful leadership begins with mastering the managerial role. PMID- 12655892 TI - Improving emergency department services. Ball Memorial strives for a more patient friendly Emergency Department. PMID- 12655894 TI - Use these tools to comply with patient safety goals. AB - Technology can help you comply with the 2003 National Patient Safety Goals from the Joint Commission on Accreditation of Healthcare Organizations. EDs are beginning to invest in these resources: Bar-coding systems can ensure that patients receive the correct medication. Software can alert physicians when nonstandard doses are ordered. Computerized order entry can give correct doses if you enter the patient's weight. PMID- 12655893 TI - Are pediatric drug errors occurring in your ED? Act now before tragedy strikes. AB - Children are at high risk for errors in the ED, and the mistakes often are due to weight-based dosing errors. Reduce the number of drugs, concentrations, formulations, and suppliers used. Have ED prescriptions reviewed by a pharmacist. Have two nurses or physicians verify dosages for high-risk drugs such as sedating agents. PMID- 12655895 TI - Speed up orders for inpatients held in ED. AB - You need a policy to ensure that important orders are carried out for admitted patients being held in the ED. ED physicians should not succumb to pressure to write inpatient orders. Transitional orders can be used for stable inpatients during their stay in the ED. By using transitional orders, inpatients being held in the ED can be sent upstairs more quickly. PMID- 12655896 TI - Deluged with inpatient holds? Avoid violations. AB - To comply with standards of the Joint Commission on Accreditation of Healthcare Organizations, you must provide the same level of care to inpatients held in your ED as they would receive in critical care units. Train the entire staff in telemetry monitoring. Give patients the same level of services from radiology, laboratory, and pharmacy as they would receive upstairs. Have ED nurse work occasional shifts in critical care units. PMID- 12655897 TI - Work closely with coders or ED revenue may be lost. PMID- 12655898 TI - Physicians insist on sending their patients to our ED for injections to treat ongoing medical conditions. PMID- 12655899 TI - A broad cast of network players. PMID- 12655900 TI - Perspectives from sterile processing and perioperative services. PMID- 12655901 TI - Performance lines in 2003. PMID- 12655902 TI - Measurable differences. PMID- 12655903 TI - Quality of life in OCD: differential impact of obsessions, compulsions, and depression comorbidity. AB - OBJECTIVE: An anxiety disorder severely affects the sufferer's quality of life (QOL), and this may be particularly true of those with obsessive-compulsive disorder (OCD). This study examines the differential impact of obsessions, compulsions, and depression comorbidity on the QOL of individuals with OCD. METHOD: Forty-three individuals diagnosed with OCD according to DSM-IV criteria and experiencing clinically significant obsessions and compulsions completed measures of QOL, obsessive-compulsive symptom severity, and depression severity. RESULTS: Obsession severity was found to significantly predict patient QOL, whereas the severity of compulsive rituals did not impact on QOL ratings. Comorbid depression severity was the single greatest predictor of poor QOL, accounting for 54% of the variance. CONCLUSIONS: Given the importance of these symptoms, treatments that directly target obsessions and secondary depression symptoms in OCD are warranted. However, replication of these findings in a prospective cohort study is required, because although the the current study's cross-sectional design allows for the examination of the associations among obsessions, depression, and QOL, it cannot establish their temporal framework (that is, causal relations). PMID- 12655904 TI - A pilot study of a parent-education group for families affected by depression. AB - OBJECTIVE: This study assessed the feasibility and efficacy of a parent-education group for families with young children and a parent with depression. We designed the program to be readily disseminated if shown to be effective. METHOD: We recruited 44 parents with depression from clinics and family doctors in Hamilton, Ontario, and randomly assigned them to receive the parenting program or to a wait list control group. The outcomes measured included knowledge of depression, parenting, family relationships, depression symptoms, child depressive symptoms, and functioning. We used analysis of covariance to test for posttreatment differences between experimental and control groups. RESULTS: Of the treatment group, 27% dropped out at posttreatment, and 43% by follow-up. Those who dropped out had more severe depression at baseline than did those who completed the program, and there was selective loss of parents with more severe depression in the experimental group. In intention-to-treat analyses at posttreatment, probands in the experimental group reported more improvements on family functioning, parenting sense of competence, and family and parent conflict than did control subjects. Standardized effect sizes (ES) were medium (0.4 to 0.6). When baseline depressive symptom scores were controlled in the analyses, the between-group differences were reduced, showing that selective loss of participants may have influenced the findings. CONCLUSIONS: On balance, the results are encouraging and support the further development and evaluation of the group intervention. However, the study does not provide unequivocal evidence in support of the program. Before it is transferred to other settings, the program needs further modification to improve participation by parents with more severe depression and further evaluation of its effectiveness. PMID- 12655905 TI - Differentiating symptoms of complicated grief and depression among psychiatric outpatients. AB - OBJECTIVE: This study examined whether dimensions of complicated grief (CG) could be distinguished from dimensions of depression and whether these dimensions were differentially affected by group psychotherapy for CG. METHOD: A total of 398 psychiatric outpatients who had experienced one or more significant death losses provided ratings on standard measures of grief and depression. Factor analysis of the 56 items from these measures was used to explore the possibility that grief and depression symptoms would form separate dimensions of distress. Subsamples of the patients also participated in 1 of 2 forms of short-term group therapy for CG. Repeated-measures analysis of variance and calculation of effect sizes were performed to examine changes in the dimensions following treatment. RESULTS: The grief items formed 3 distinct clusters representing different dimensions of CG. None of the depression items loaded highly on these grief dimensions. The depression items formed 2 distinct clusters. Two of the grief dimensions demonstrated the most improvement following group therapy that addressed CG. There was also evidence for differential effectiveness of the 2 forms of group therapy. CONCLUSIONS: When assessing psychiatric patients who have death losses, clinicians should consider different types of grief reactions. Different types of grief reactions may be responsive to different treatments. In the absence of depressive symptoms, clinicians should not assume the absence of CG.